PMID- 14519407 TI - Web-based remote monitoring of infant incubators in the ICU. AB - A web-based real-time operating, management, and monitoring system for checking temperature and humidity within infant incubators using the Intranet has been developed and installed in the infant Intensive Care Unit (ICU). We have created a pilot system which has a temperature and humidity sensor and a measuring module in each incubator, which is connected to a web-server board via an RS485 port. The system transmits signals using standard web-based TCP/IP so that users can access the system from any Internet-connected personal computer in the hospital. Using this method, the system gathers temperature and humidity data transmitted from the measuring modules via the RS485 port on the web-server board and creates a web document containing these data. The system manager can maintain centralized supervisory monitoring of the situations in all incubators while sitting within the infant ICU at a work space equipped with a personal computer. The system can be set to monitor unusual circumstances and to emit an alarm signal expressed as a sound or a light on a measuring module connected to the related incubator. If the system is configured with a large number of incubators connected to a centralized supervisory monitoring station, it will improve convenience and assure meaningful improvement in response to incidents that require intervention. PMID- 14519406 TI - Primary care physician attitudes concerning follow-up of abnormal test results and ambulatory decision support systems. AB - OBJECTIVES: Failures to follow-up abnormal test results are common in ambulatory care. Information systems could assist providers with abnormal test result tracking, yet little is known about primary care providers attitudes toward outpatient decision support systems. METHODS: A cross-sectional survey of 216 primary care physicians (PCPs) that utilize a single electronic medical record (EMR) without computer-based clinical decision support. RESULTS: The overall response rate was 65% (140/216). Less than one-third of the respondents were satisfied with their current system to manage abnormal laboratory, radiographs, Pap smear, or mammograms results. Only 15% of providers were satisfied with their system to notify patients of abnormal results. Over 90% of respondents felt automated systems to track abnormal test results would be useful. Seventy-nine percent of our respondents believed that they could comply better with guidelines through electronic clinical reminders. CONCLUSIONS: Most PCPs were not satisfied with their methods for tracking abnormal results. Respondents believed that clinical decision support systems (CDSS) would be useful and could improve their ability to track abnormal results. PMID- 14519408 TI - Wireless and PDA: a novel strategy to access DICOM-compliant medical data on mobile devices. AB - The medical procedures at the patient bedside are out of the scope of current HIS/RIS systems, which means that patient record and image data access during the medical visit or the execution, recording and confirmation of the medicine prescriptions, still do not enjoy computerized support. As a consequence, the necessary inclusion of new data to the patient record, still needs to be carried out through notations on paper and later typed, causes delays on the availability of this information (Mobile computing in a hospital: the Ward-In-Hand project. SAC 2000-ACM Symposium on Applied Computing, Marco, 2000, Villa Olmo, Como, Italy). This paper presents a software technology where the medical and nursing staff will be equipped with a handheld computer connected by radio to a central server that provides access to the electronic patient records and that will actively inform about tasks pending distribution. The server acts as a database for multimodal electronic patient record information, storing patient data from computerized tomography (CT), ultrasonography (US), magnetic resonance (MRI) images and also medical findings, observations and prescriptions coded as DICOM Structured Reports (Digital Image and Communications in Medicine). The prototype described in this article implements a medical images and structured reports server that makes the search and recovery of data stored in the DICOM standard possible. PMID- 14519409 TI - Open source model for global collaboration in higher education. PMID- 14519410 TI - Binary combinations of propofol and barbiturates on human alpha(1) glycine receptors expressed in Xenopus oocytes. AB - To test whether there is a common site of action for intravenous anaesthetics at the glycine receptor, the effects of binary combinations of thiopentone, pentobarbitone, methohexitone, and propofol have been tested on human alpha(1) glycine receptors expressed in Xenopus laevis oocytes using two-electrode voltage clamp techniques. Thiopentone (5-40 microM), pentobarbitone (25-400 microM) and propofol (2-100 microM) (but not methohexitone), potentiated the glycine-induced (50 microM) current in a dose-dependent manner, with the maximum potentiation observed to be 218%, 400%, and 576%, respectively. In binary combination with thiopentone, pentobarbitone or propofol, methohexitone reduced potentiation compared to that by the individual anesthetics to 190%, 260% and 460%, respectively. Combination of thiopentone and pentobarbitone (50 microM) increased potentiation, compared to that by thiopentone alone. Binary combinations of propofol with either thiopentone or pentobarbitone showed more potentiation, compared to that observed with the individual anesthetics. Our results indicate that thiopentone, pentobarbitone and propofol all act as positive allosteric modulators at the alpha(1) glycine receptor. In contrast, methohexitone has no action alone but acts as a competitive antagonist to thiopentone, pentobarbitone and propofol. We suggest that, on the basis of these results, these four intravenous anaesthetics share a common site of action at the glycine receptor. PMID- 14519411 TI - Possible antioxidant and neuroprotective mechanisms of FK506 in attenuating haloperidol-induced orofacial dyskinesia. AB - Tardive dyskinesia is a serious motor side effect of chronic neuroleptic therapy. The pathophysiology of this disabling and commonly irreversible movement disorder is still obscure. It may be caused by a loss of dopaminergic cells, due to free radicals as a product of high synaptic dopamine levels. Chronic treatment with neuroleptics leads to the development of abnormal oral movements in rats called vacuous chewing movements. Vacuous chewing movements in rats are widely accepted as an animal model of tardive dyskinesia. Chronic haloperidol (1 mg/kg for 21 days) treatment significantly induced vacuous chewing movements and tongue protrusions in rats, and FK506 (Tacrolimus) [[3S [3R*[E(1S*,3S*,4S*)],4S*,5R*,8S*,9E,12R*,14R*,15S*,16R*,18S*,19S*,26aR*]] 5,6,8,11,12,13,14,15,16,17,18,19,24,25,26,26a-hexadecahydro-5, 19-dihydroxy-3-[2 (4-hydroxy-3-methoxycyclohexyl)-1-methylethenyl]-14, 16-dimethoxy-4,10,12, 18 tetramethyl-8-(2-propenyl)-15, 19-epoxy-3H-pyrido[2,1-c][1,4] oxaazacyclotricosine-1,7,20, 21(4H,23H)-tetrone, monohydrate] dose dependently (0.5 and 1 mg/kg) reduced these haloperidol-induced movements. Biochemical analysis revealed that chronic haloperidol treatment significantly induced lipid peroxidation and decreased the levels of glutathione and of the antioxidant defense enzymes, superoxide dismutase and catalase, in the brains of rats. Co administration of FK506 dose dependently (0.5 and 1 mg/kg) and significantly reduced the lipid peroxidation and restored the decreased glutathione levels induced by chronic haloperidol treatment. It also significantly reversed the haloperidol-induced decrease in brain superoxide dismutase and catalase levels. The major findings of the present study suggest that oxidative stress-induced neuronal death might play a significant role in neuroleptic-induced orofacial dyskinesia. In conclusion, FK506 could be a useful drug for the treatment of neuroleptic-induced orofacial dyskinesia. PMID- 14519412 TI - Inducible nitric oxide synthase is involved in corticotropin-releasing hormone mediated central sympatho-adrenal outflow in rats. AB - Brain nitric oxide (NO), recognized as a neurotransmitter or a neuromodulator, is mainly generated either by neuronal NO synthase (NOS) or by inducible NOS. NO has been shown to activate cyclooxygenase (a prostaglandin-forming enzyme) in addition to guanylate cyclase. Recently, we reported that the intracerebroventricularly (i.c.v.) administered corticotropin-releasing hormone (CRH) increases plasma catecholamines through brain cyclooxygenase-dependent mechanisms in rats. In the present experiments, therefore, we examined whether NO is involved in the CRH-induced increase of plasma catecholamines using urethane anesthetized rats. I.c.v. administered CRH increased plasma noradrenaline and adrenaline in a dose-dependent manner (0.5, 1.5, and 3.0 nmol/animal). The CRH (1.5 nmol/animal, i.c.v.)-induced increase of plasma catecholamines was reduced by N(omega)-nitro-L-arginine methyl ester (a non-selective inhibitor of NOS) [111 nmol (30 microg)/animal, i.c.v.], but not by the same dose of N(omega)-nitro-D arginine methyl ester (an inactive isomer of N(omega)-nitro-L-arginine methyl ester). The CRH-induced increase of plasma catecholamines was also reduced either by cycloheximide (an inhibitor of protein synthesis) [107 nmol (30 microg)/animal, i.c.v.] or by S-methylisothiourea (an inhibitor of inducible NOS) [71 nmol (20 microg) and 711 nmol (200 microg)/animal, i.c.v.]. These results suggest the involvement of brain inducible NOS in the CRH-induced activation of the central sympatho-adrenomedullary outflow in rats. PMID- 14519413 TI - Differential effects of antipsychotics on haloperidol-induced vacuous chewing movements and subcortical gene expression in the rat. AB - The behavioral and neurochemical effects of switching from typical to atypical medications have not been evaluated in the rodent models of tardive dyskinesia. Thus, we treated rats with haloperidol-decanoate for 12 weeks, and assessed the effects of additional treatment with olanzapine, haloperidol, clozapine, or vehicle on vacuous chewing movements and expression of transcripts for dopamine receptors, tyrosine hydroxylase, delta-opioid receptor, prodynorphin, preproenkephalin, glutamic acid decarboxylase-65 (glutamic acid decarboxylase (GAD)-65) and GAD-67 and N-methyl-D-aspartate (NMDA) receptor subunits in the striatum and its efferent pathways. Haloperidol-decanoate induced vacuous chewing movements extinguished following an additional 4 weeks of treatment with vehicle, olanzapine or haloperidol, but not clozapine. Post-treatment, vacuous chewing movements in the clozapine group were significantly higher than the vehicle, olanzapine and haloperidol groups. GAD-67 mRNA expression in the globus pallidus was decreased following additional treatment with olanzapine or haloperidol, but not clozapine. Changes in expression of other transcripts were not detected. These findings demonstrate important differences in the effects of typical and atypical antipsychotics on chronic vacuous chewing movements. PMID- 14519414 TI - Gradient of dopamine responsiveness to dopamine receptor agonists in subregions of the rat nucleus accumbens. AB - The present study sought to investigate the possibility that the degree of selectivity of dopamine D3/D2 receptor agonists such as quinelorane, 7-hydroxy-2 dipropylaminotetralin (7-OH-DPAT), quinpirole and apomorphine on dopamine D3 over D2 receptor subtypes can be assessed by measuring dopamine transmission in the shell vs. core compartments of the nucleus accumbens by using microdialysis in freely moving rats. Significant reductions in dialysate dopamine levels compared to vehicle-treated animals were observed in the shell of the nucleus accumbens with 3, 10 and 30 microg/kg quinelorane, 100 microg/kg 7-OH DPAT, 25 and 100 microg/kg quinpirole, and 100 microg/kg apomorphine. In the core subregion, significant reductions in dopamine were seen at 10 and 30 microg/kg quinelorane, 25 and 100 microg/kg 7-OH-DPAT, 100 microg/kg quinpirole and 100 microg/kg apomorphine. However, a significant shell/core dichotomy could only be observed in response to the lowest dose of quinelorane (3 microg/kg) with the shell being hyper-responsive compared with the core. The present findings suggest that quinelorane is one of the most selective dopamine D3 receptor agonists based on its ability to target the shell subregion of the nucleus accumbens. PMID- 14519415 TI - Effects of alpha(1)-adrenoceptors and muscarinic cholinoceptors on water intake in rats. AB - The present study investigated the effects of alpha(1)-adrenoceptors and muscarinic cholinoceptors on water intake in adult male rats. Intracerebroventricular (i.c.v.) injections were carried out in all experiments after 24-h deprivation of water. After deprivation, the volume of consumed water was measured for 1 h. Administration of pilocarpine, a muscarinic cholinoceptor agonist (0.5-1 microg/rat), and prazosin, the alpha(1)-adrenoceptors antagonist (2 microg/rat), increased, while scopolamine, a muscarinic cholinoceptor antagonist (5-10 microg/rat), and phenylephrine, an alpha(1)-adrenoceptor agonist (30 microg/rat), decreased water intake in rats. The activation of muscarinic cholinoceptors by pilocarpine attenuated the inhibitory effect induced by phenylephrine. Blockade of muscarinic cholinoceptors did not change the phenylephrine-induced response. Pretreatment with prazosin decreased the pilocarpine-induced response. However, pharmacological blockade of muscarinic cholinoceptors by scopolamine decreased the prazosin-induced effect on water intake. It is concluded that muscarinic cholinoceptors and alpha(1)-adrenoceptors may interact on water intake. PMID- 14519416 TI - Very low doses of ethanol induce behavioral changes involving dopamine D2 receptors. AB - In male rats, pretreatment for 20 days with very low (0.5, 1%, v/v) but not with high (5, 10%, v/v) oral doses of ethanol delayed the initiation and reduced the duration of narcosis induced by an acute high intraperitoneal (i.p.) dose of the drug (3 g/kg in 25% saline solution). Furthermore, the treatment improved the acquisition of shuttle-box active avoidance response but did not affect the emission of ultrasonic calls, an index of emotional state of animals. These effects were inhibited by peripheral administration of the dopamine D2 receptor antagonist, sulpiride (1 mg/kg). A higher dose of sulpiride (10 mg/kg) prolonged the duration of narcosis in rats pretreated with high-dose ethanol and reduced the number of conditioned avoidance responses in the shuttle-box paradigm. The pretreatment with the dopamine D2 receptor agonist, (+/-)-2-(N-phenethyl-N propylamino)-5-hydroxytetralin (PPHT, 0.1 mg/kg), enhanced the effects of ethanol very low doses in delaying the initiation and reducing the duration of narcosis induced by an acute i.p. dose of the drug. A pharmacokinetic study in ethanol pretreated animals revealed that administration of 0.5% or 1% ethanol for 20 days did not modify significantly the bioavailability of acute ethanol administered i.p. in a dose of 3 g/kg in 25% saline solution. Thus, repeated administration of ethanol very low doses may have affected the sensitivity of presynaptic dopamine D2 receptors. The influence on dopamine release through an action on presynaptic receptors may be involved in these effects of small doses of ethanol. PMID- 14519417 TI - Effects of alpha(1)-acid glycoprotein on isometric tension of mouse aorta. AB - We examined the effects of human alpha(1)-acid glycoprotein on isometric tension of mouse aortic rings. alpha(1)-Acid glycoprotein (7.5-75 microM) produced a transient, concentration-dependent relaxation of the phenylephrine-precontracted preparation. Although N(G)-nitro-L-arginine methyl ester or removal of endothelium rarely affected the alpha(1)-acid glycoprotein-induced relaxation, extracellular heparin inhibited the alpha(1)-acid glycoprotein-induced relaxation. In 10 mM Ca(2+)-containing external solutions, the alpha(1)-acid glycoprotein-induced relaxation was significantly potentiated. In the 60 mM KCl precontracted preparation, alpha(1)-acid glycoprotein produced weaker relaxation than in the phenylephrine-precontracted preparation. These results suggest that the vasorelaxant effect of alpha(1)-acid glycoprotein is mainly achieved by block of Ca(2+) entry in the vascular smooth muscle cells. The interaction between alpha(1)-acid glycoprotein molecules and plasmalemmal Ca(2+) entry channels may be modified by extracellular Ca(2+) and heparin. PMID- 14519418 TI - Age-dependent changes in blood pressure and arterial reactivity in obese Zucker rats. AB - The purpose of the present investigation was to determine whether there is an association between changes in arterial reactivity to vasoactive agents and the development of hypertension in obese Zucker rats. At 20 weeks of age, obese rats were mildly hypotensive compared to their lean littermate controls. Maximum contractile responses of endothelium-intact mesenteric arteries from these rats to noradrenaline, endothelin-1 and KCl were depressed, although there was no change in relaxation to acetylcholine. By 32 weeks of age, obese rats had developed hypertension compared to their lean littermate controls. Maximum contractile responses of mesenteric arteries from 32-week-old obese rats to noradrenaline and endothelin-1 were no longer significantly different than control, although contractile responses to KCl remained depressed. In addition, there was a small increase in sensitivity to endothelin-1, while endothelium dependent relaxation to acetylcholine was impaired. In contrast, there were no changes in contractile responses of endothelium-intact aortas from either 20- or 32-week-old obese rats to noradrenaline, endothelin-1 or KCl, while endothelium dependent relaxation of this artery to acetylcholine was slightly enhanced at both ages. Therefore, changes in the reactivity of the mesenteric artery but not the aorta from obese Zucker rats parallel changes in blood pressure in these animals. PMID- 14519419 TI - Alterations in pulmonary vascular function in rats exposed to intermittent hypoxia. AB - Vasoactive agents were examined in arteries from control rats and rats exposed to intermittent hypoxia (10% oxygen; 8 h/day) for 3, 5 or 20 days. Hypoxic rats developed right ventricular hypertrophy after 5 days, but became pulmonary hypertensive (elevated right ventricular systolic pressure; RVSP) only after 20 days. In pulmonary arteries (main and intralobar), responses to acetylcholine and ionomycin (endothelium-dependent vasodilators) were reduced after 20 and 5 days of intermittent hypoxia, whereas contractions to 5-hydroxytryptamine (5-HT) were enhanced (potency increase >10-fold) after 20, 5 and 3 days. Contractions to endothelin-1 and a thromboxane-mimetic, but not Ca(2+), were also increased. No changes in vascular function occurred in aorta. Since changes in pulmonary vascular function preceded the increase in RVSP they do not result from, but may contribute to, the development of hypoxia-induced pulmonary hypertension. If similar changes occur in humans, they may be important in conditions characterised by intermittent, as opposed to continuous, hypoxia. PMID- 14519420 TI - Relaxing effects induced by the soluble guanylyl cyclase stimulator BAY 41-2272 in human and rabbit corpus cavernosum. AB - 5-Cyclopropyl-2-[1-(2-fluoro-benzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-pyrimidin-4 ylamine (BAY 41-2272) is a potent soluble guanylyl cyclase stimulator in a nitric oxide (NO)-independent manner. The relaxant effect of BAY 41-2272 was investigated in rabbit and human corpus cavernosum in vitro. BAY 41-2272 (0.01-10 microM) relaxed both rabbit (pEC(50)=6.82+/-0.06) and human (pEC(50)=6.12+/-0.10) precontracted cavernosal strips. The guanylyl cyclase inhibitor (ODQ, 10 microM) caused significant rightward shifts in the concentration-response curves for BAY 41-2272 in rabbit (4.7-fold) and human (2.3-fold) tissues. The NO synthesis inhibitor (N-nitro-L-arginine methyl ester (L-NAME), 100 microM) also produced similar rightward shifts, revealing that BAY 41-2272 acts synergistically with endogenous NO to elicit its relaxant effect. The results also indicate that ODQ is selective for the NO-stimulated enzyme, since relaxations evoked by BAY 41 2272 were only partly attenuated by ODQ. The present study shows that both BAY 41 2272 and sildenafil evoke relaxations independent of inhibition of haem in soluble guanylate cyclase. Moreover, there is no synergistic effect of the two compounds in corpus cavernosum. PMID- 14519421 TI - Phosphodiesterase type 5 inhibition ameliorates nephrotoxicity induced by cyclosporin A in spontaneous hypertensive rats. AB - Our recent study suggests that there is a reciprocal mechanism to maintain cGMP content, via both a decrease in cGMP degradation (decrease in cGMP phosphodiesterase activity) and an increase in synthesis of cGMP (increase in guanylate cyclase activity) in the kidney of cyclosporin A-treated rats. We undertook this study to clarify the role of cGMP-phosphodiesterase in cyclosporin A nephrotoxicity by evaluating N-(3,4-dimethoxybenzyl)-2-[[(1R)-2-hydroxy-1 methylethyl]amino]-5-nitrobenzamide (FR226807), a phosphodiesterase type 5 inhibitor, in an animal model. Male spontaneous hypertensive rats (SHR) were treated with cyclosporin A (50 mg/kg) for 2 weeks or with cyclosporin A and FR226807 (3.2 mg/kg or 10 mg/kg) for 2 weeks. Cyclosporin A-treated rats showed renal dysfunction and histological change compared with vehicle-treated rats. Administration of FR226807 improved the renal dysfunction (increase in serum creatinine and fractional excretion of sodium, and decrease in creatinine clearance) as well as the pathological changes (tubular vacuolization) induced by cyclosporin A in SHR. At the molecular level, administration of FR226807 resulted in a further increase in cGMP content in the kidney, aorta and platelets from cyclosporin A-treated rats. Our present study demonstrates that cGMP phosphodiesterase plays an important role in the cyclosporin A nephrotoxicity and also suggests that further inhibition of cGMP-phosphodiesterase is a potential pharmacological target for preventing cyclosporin A nephrotoxicity. PMID- 14519422 TI - Rosiglitazone-induced protection against myelotoxicity produced by 5 fluorouracil. AB - Insulin promotes survival of haemopoietic progenitors. We investigated if rosiglitazone, an insulin sensitizer, could confer protection against 5 fluorouracil (5-FU)-induced myelotoxicity in mice. The decrease in bone marrow cellularity, frequency and content of granulocyte-macrophage progenitors (CFU-GM) characterized myelotoxicity in mice, while insulin sensitivity was determined by hyperinsulinaemic euglycaemic glucose clamping. CFU-GM colony numbers increased in groups pre-treated with rosiglitazone (1.5-6 mg/kg, 5 days), compared to that in mice treated with 5-fluorouracil alone. Since rosiglitazone pre-treatment significantly promoted the clonal expansion of CFU-GM when given in the insulin sensitizing dose, we conclude that rosiglitazone had myeloprotective effects possibly by amplifying endogenous insulin action. PMID- 14519424 TI - The beta-adrenergic receptor kinase in heart failure. AB - Heart failure (HF) remains a significant and increasing cause of worldwide morbidity and mortality. HF is less a disease than a common clinical endpoint resulting from diverse, but often co-existing etiologies-including hypertension, coronary artery disease, and viral cardiomyopathy. Regardless of the pathologic trigger, HF can be characterized by a series of specific, molecular changes in the diseased myocardium. Noteworthy among these changes are alterations in the beta-adrenergic receptor (betaAR) signaling cascade. betaARs belong to the larger family of G-protein-coupled receptors (GPCRs) and modulate cardiac function by controlling the inotropic and chronotropic response to catecholamines. betaARs, in turn, are regulated by GPCR kinases (GRKs). GRKs phosphorylate betaARs, blocking downstream-signaling cascades and ultimately desensitizing the receptor to further catecholamine stimuli. Recent advances in transgenic mouse and gene therapy techniques have led to therapeutic strategies by manipulating betaAR signaling, specifically through the inhibition of the beta-adrenergic receptor kinase (betaARK1 or GRK2), the predominant myocardial GRK. The purpose of this manuscript, then, is to review (1). the changes that occur to betaAR-signaling pathways in HF, (2). the evidence from transgenic murine studies examining the consequences of betaARK1 manipulation in the failing heart, and (3). the effectiveness of in vivo applications of betaARK1-targeted gene therapy at ameliorating HF. PMID- 14519423 TI - Propofol ameliorates liver dysfunction and inhibits aortic superoxide level in conscious rats with endotoxic shock. AB - Propofol, widely used as a sedative agent, is known to exert antioxidant and anti inflammatory effects in vitro. We studied the effects of propofol on hemodynamics and the function of several organs in conscious rats with endotoxemia. Intravenous injection of rats with endotoxin (lipopolysaccharide) caused hypotension, vascular hyporeactivity and tachycardia as well as significant lung, liver and kidney damage. Hepatocellular damage caused by lipopolysaccharide for 6 h was significantly attenuated in the lipopolysaccharide+propofol group. Aortic superoxide anion (O(2)(radical)(-)) production, but not plasma nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) level, was also suppressed by propofol in lipopolysaccharide-injected rats. Light microscopy showed that propofol attenuated the marked infiltration of neutrophils in liver tissues from lipopolysaccharide-injected rats. Moreover, the survival rate of the lipopolysaccharide+propofol group at 16 h was significantly increased when compared with that of the lipopolysaccharide group (53% vs. 12%). These results suggest that inhibition of aortic O(2)(radical)(-) production and amelioration of liver dysfunction contribute to the beneficial effect of propofol in conscious rats with endotoxic shock. PMID- 14519425 TI - TBX5: a developmental key that fits many locks. PMID- 14519426 TI - Complexity of transcriptional regulation of the tumor necrosis factor-alpha gene. PMID- 14519427 TI - alpha(1)-adrenoceptors in the heart - friend or foe? PMID- 14519428 TI - How glucose and glucose transporters protect cardiac myocytes. PMID- 14519429 TI - TBX5 nuclear localization is mediated by dual cooperative intramolecular signals. AB - The TBX5 transcription factor is required for normal cardiogenesis, and human TBX5 mutations cause congenital heart defects. Previous studies have shown that TBX5 can localize to cellular nuclei during embryogenesis and have suggested that altered nuclear localization may contribute to disease pathogenesis. Current analyses suggest that TBX5 nuclear localization is not uniform during organogenesis. To determine the biochemical mechanisms underlying TBX5 nuclear import, we performed site-directed mutagenesis of human TBX5. We identified two distinct nuclear localization signals in TBX5, one monopartite and one bipartite. While each is insufficient to promote complete TBX5 nuclear localization, they act cooperatively to do so. These sequences are evolutionarily conserved and have cognates in other T-box gene family members. PMID- 14519430 TI - Regulation of the human tumor necrosis factor-alpha promoter by angiotensin II and lipopolysaccharide in cardiac fibroblasts: different cis-acting promoter sequences and transcriptional factors. AB - We recently showed that angiotensin (ANG) II as well as mechanical stretch stimulated production of tumor necrosis factor (TNF) in cardiac fibroblasts. Presently, we examined the molecular mechanisms by which ANGII and lipopolysaccharide (LPS) upregulate TNF-alpha gene expression. In neonatal rat cardiac fibroblasts, increased transcription of TNF-alpha mRNA was detected as luciferase activity associated with activity of the TNF-alpha promoter. Progressive deletion from this promoter located the LPS-responsive region between -200 and -120 bp from the transcription initiation site, while the sequence between -120 and -70 bp was required for ANGII-induced expression. Next, we examined which cis-acting sequences in the TNF-alpha promoter region were essential for induction of TNF-alpha transcription. Competition analysis by electrophoretic mobility shift assay with and without specific antibodies showed that LPS increased binding of Sp1 and Sp3 to the Sp1-binding site, while Egr-1 was unimportant. With ANGII, binding of ATF-2/c-jun to the CRE site was required for TNF-alpha gene induction; neither Ets nor NF-kappaB was essential. Mutation analysis confirmed that response to LPS relied upon the Sp1 site in the TNF-alpha promoter, while the CRE-binding site was essential for stimulation by ANGII. We concluded that since TNF-alpha gene expression is transcriptionally activated by ANGII or LPS in cardiac fibroblasts via different cis-acting sequences in the TNF alpha promoter and different transcriptional factors, mechanisms inducing TNF production differ between heart failure or cardiac hypertrophy and infectious disease. PMID- 14519431 TI - Abnormal myocardial contraction in alpha(1A)- and alpha(1B)-adrenoceptor double knockout mice. AB - We used double-knockout mice (ABKO) lacking both predominant myocardial alpha(1) adrenergic receptor (AR) subtypes (alpha(1A) and alpha(1B)) to determine if alpha(1)-ARs are required for normal myocardial contraction. Langendorff-perfused ABKO hearts had higher developed pressure than wild type (WT) hearts (123 +/- 3 mmHg n = 22 vs. 103 +/- 3 mmHg, n = 38, P < 0.001). Acutely inhibiting alpha(1) ARs in WT hearts with prazosin did not increase pressure, suggesting that the increased pressure of ABKO hearts was mediated by long-term trophic effects on contraction rather than direct regulatory effects of alpha(1)-AR removal. Similar to perfused hearts, ABKO ventricular trabeculae had higher submaximal force at 2 mM extracellular [Ca(2+)] than WT (11.4 +/- 1.7 vs. 6.9 +/- 0.6 mN/mm(2), n = 8, P < 0.05); however, the peaks of fura-2 Ca(2+) transients were not different (0.79 +/- 0.11 vs. 0.75 +/- 0.16 microM, n = 10-12, P > 0.05), suggesting ABKO myocardium had increased myofilament Ca(2+)-sensitivity. This conclusion was supported by measuring the Ca(2+)-force relationship using tetanization. Increased myofilament Ca(2+)-sensitivity was not explained by intracellular pH, which did not differ between ABKO and WT (7.41 +/- 0.01 vs. 7.39 +/- 0.02, n = 4 6, P > 0.05; from BCECF fluorescence). However, ABKO displayed impaired troponin I phosphorylation, which may have played a role. In contrast to increased submaximal force, ABKO trabeculae had lower maximal force than WT at high extracellular [Ca(2+)] (29.6 +/- 1.9 vs. 37.6 +/- 1.4 mN/mm(2), n = 7, P < 0.01). However, peak cytosolic [Ca(2+)] was not different (1.13 +/- 0.15 vs. 1.19 +/- 0.04 microM, n = 6-7, P > 0.05), suggesting ABKO myocardium had impaired myofilament function. Finally, ABKO myocardium had decreased responsiveness to beta-AR stimulation. We conclude: alpha(1)-ARs are required for normal myocardial contraction; alpha(1)-ARs mediate long-term trophic effects on contraction; loss of alpha(1)-AR function causes some of the functional abnormalities that are also found in heart failure. PMID- 14519432 TI - Upregulation of GLUT1 expression is necessary for hypertrophy and survival of neonatal rat cardiomyocytes. AB - During hypertrophy the heart increases its utilization of glucose and decreases that of fatty acids, resuming a fetal pattern of substrate metabolism. As demonstrated here, GLUT1 protein expression is increased in association with in vivo pressure-overload-induced hypertrophy. The relationship of changes in GLUT1 to enhanced glucose uptake and to cardiomyocyte hypertrophy and survival is not known. To explore this question we first examined the effect of prostaglandin F2alpha (PGF2alpha), an established hypertrophic agonist, on GLUT1 expression and glucose uptake in neonatal rat ventricular myocytes (NRVMs). PGF2alpha treatment for 24 h led to a fivefold increase in GLUT1 expression and a sixfold increase in glucose uptake. However, NRVMs cultured in the absence of glucose or with 3-O methyl glucose, a competitive inhibitor of glucose uptake, still exhibited PGF2alpha-induced hypertrophic growth. In addition, we determined that overexpression of GLUT1 using adenovirus was insufficient to cause an increase in cell size, myofibrillar organization, or atrial natriuretic factor (ANF) expression. On the other hand, adenoviral overexpression of antisense GLUT1 (which blocked PGF2alpha-induced increases in GLUT1 protein) prevented PGF2alpha stimulated cell enlargement and increases in ANF transcription. Overexpression of GLUT1 or addition of PGF2alpha also protected cells against serum deprivation induced apoptosis; this effect was blocked by antisense GLUT1 but, surprisingly, was not dependent on glucose. Together, these data suggest that upregulation of GLUT1 serves a role in agonist-induced hypertrophy and survival which can be dissociated from its role in glucose transport. PMID- 14519433 TI - Tyrosine kinases inhibitors block Fas-mediated deleterious effects in normoxic and hypoxic ventricular myocytes. AB - Experimental evidences suggest an important role for Fas receptor activation in a wide range of myocardial pathologies, which are associated with a variety of arrhythmias and mechanical dysfunction. Our recent studies have shown that Fas mediated arrhythmias and mechanical disturbances of ventricular myocytes can be accounted for by activation of the phospholipase C-1,4,5-inositol triphosphate intracellular calcium release (PLC-1,4,5-IP(3)-[Ca(2+)](i)) cascade, which is responsible for attenuating the transient outward current (I(to)) and augmenting the L-type Ca(2+) current (I(Ca,L)). We have also shown that whereas ventricular myocytes are resistant to Fas-mediated apoptosis, hypoxia predisposes myocytes to apoptosis induced by Fas activation by shifting the balance between pro-apoptotic and anti-apoptotic proteins towards the former. Since protein tyrosine phosphorylation has been shown to be involved in Fas signaling, we have hypothesized that inhibiting tyrosine kinases will attenuate Fas-mediated effects in ventricular myocytes in normoxic and hypoxic conditions. Therefore, we tested the effect of the tyrosine kinases inhibitors, genistein (50 micromol/l) and herbimycin A (50 microg/ml), on the abovementioned Fas-mediated effects in cultured neonatal rat ventricular myocytes (NRVM) and in freshly isolated adult murine ventricular myocytes. Fas receptor was activated by incubating NRVM with recombinant Fas ligand (rFasL, 50 ng/ml) and enhancing antibody (1 microg/ml), or by incubation of murine ventricular myocytes with the Fas-activating antibody Jo2 (10 microg/ml). The major findings were that genistein prevented Fas-mediated increase in 1,4,5-IP(3) production in NRVM (quantified by ion-exchange chromatography): 216 +/- 41 counts per minute (cpm) in control, 605 +/- 184 cpm in FasL-treated cardiomyocytes and 137 +/- 51 cpm in rFasL + genistein-treated cultures. Accordingly, genistein or herbimycin A abolished the diastolic [Ca(2+)](i)-rise (as measured by fura-2 fluorescence) and arrhythmogenic activity in both rat and murine ventricular myocytes, and the Fas-mediated changes in I(to) and I(Ca,L) in murine ventricular myocytes. Importantly, genistein attenuated Fas-mediated apoptosis in hypoxic (22 h in 1% O(2)) NRVM; the apoptotic ratios as measured by DAPI fluorescence assay were: 4.6 +/- 1.0% in control, 12.5 +/- 3.0% in rFasL and 7.3 +/- 1.6% in rFasL + genistein-treated NRVM. This prevention of apoptosis by tyrosine kinases blockade was accompanied by inhibition of hypoxia-induced increased Fas expression and decreased expression of the anti-apoptotic protein xIAP. In conclusion, our findings indicate that tyrosine phosphorylation is involved in Fas signaling in ventricular myocytes, and can, therefore, serve as a novel potential target for attenuating Fas-mediated dysfunction in normoxic and hypoxic myocardium. PMID- 14519434 TI - Differential coupling of m-cholinoceptors to Gi/Go-proteins in failing human myocardium. AB - Muscarinic acetylcholine receptors (mAChRs) mediate their main cardiac effects via pertussis toxin-sensitive G-proteins. Physiological effects differ considerably between atrium and ventricle, and it is unknown to which extent these differences derive from selective receptor-G-protein coupling or further downstream events. We have characterized specific coupling between mAChRs and Gi/Go-protein isoforms in atrial and ventricular myocardium by agonist-dependent photoaffinity labeling with [(32)P]azidoanilido GTP (aaGTP) and immunoprecipitation in sarcolemmal membranes from terminally failing human hearts. The total amount of mAChRs, as determined by specific binding of [(3)H]QNB, was significantly higher in right-atrial (RA +/- SEM, 959 +/- 68 fmol/mg, n = 4) than in left-ventricular membranes (LV, 582 +/- 53 fmol/mg, n = 6). Standardized immunoblots revealed that Gialpha-2 was the predominant subtype in both regions. A 40-kDa splice variant of Goalpha (Goalpha-1 and/or Goalpha-3) was almost exclusively detectable in RA. Levels of Gialpha-3 and a 39-kDa splice variant of Goalpha (Goalpha-2) were also higher in RA. Basal aaGTP binding was higher in RA than in LV for all Gialpha/Goalpha subtypes. The carbachol (10 micromol/l)-induced increase in aaGTP binding was significantly higher in RA than in LV for Goalpha-1/3 (336 +/- 95% of LV, n = 4) and for Gialpha-3 (211 +/- 83%), lower for Gialpha-2 (42 +/- 5%), and was similar in both regions for Goalpha-2 (130 +/- 62%). The differential coupling of mAChRs in human RA and LV suggests that the initiation of different physiological responses to mAChR stimulation starts with signal sorting at the receptor-G-protein level. PMID- 14519435 TI - Mutation analysis of AMP-activated protein kinase subunits in inherited cardiomyopathies: implications for kinase function and disease pathogenesis. AB - Familial hypertrophic cardiomyopathy (HCM) has been defined as a disease of the cardiac sarcomere, although sarcomeric protein mutations are not found in one third of cases. We have recently shown that HCM associated with Wolff-Parkinson White syndrome (WPW) and conduction disease can be caused by mutations in PRKAG2, which encodes the gamma2 subunit of AMPK, an enzyme central to cellular energy homeostasis. AMPK is a heterotrimer composed of one catalytic subunit (alpha) and two regulatory subunits (beta and gamma). Seven known genes encode the subunit isoforms (alpha1, alpha2, beta1, beta2, gamma1, gamma2, gamma3) and all are expressed in the heart. To better understand the role of AMPK mutations in HCM/WPW and other inherited cardiomyophathies, all 7 subunit genes were screened for mutations in a panel of probands: 3 with HCM/WPW, 4 with DCM/WPW, 38 with HCM alone (in whom contractile protein mutations had not been found) and 13 with DCM alone. In total, 73 amplimers were screened in the 58 probands and a number of polymorphisms, including non-conservative substitutions, were identified. However, no further disease-causing mutations were found in any AMPK subunit gene. These results indicate that HCM with WPW is a distinct, but genetically heterogeneous, condition caused by mutations in PRKAG2 and in an unknown gene or genes, not involved in the AMPK complex. Mutations in PRKAG2 appear to specifically cause HCM with WPW and conduction disease, and not other inherited cardiomyopathies. As deleterious alleles were not found in other AMPK subunit isoforms, the mutations affecting PRKAG2 are likely to confer a specific alteration of AMPK function of particular importance in the myocardium. PMID- 14519436 TI - Obligatory role of diastolic voltage oscillations in sino-atrial node discharge. AB - The role of diastolic voltage oscillations in the initiation and maintenance of pacemaker discharge was studied in guinea pig-isolated sino-atrial (SA) node by means of a microelecrode technique. When [K(+)](o) is suitably increased, the maximum diastolic potential decreases and all action potentials (APs) assume the characteristics of dominant pacemakers (slow responses with U-shaped diastolic depolarization). Subsequently, as the slope and amplitude of diastolic depolarization (DD) decreases, the threshold is missed, unmasking the fused oscillatory potentials V(os) and ThV(os). As high [K(+)](o) perfusion continues, the oscillatory potentials become separated, V(os) following the AP and ThV(os) appearing later on, when DD enters a less negative voltage range (oscillatory zone). ThV(os) grow in amplitude and attain the threshold, thereby insuring a slow discharge. If [K(+)](o) is further increased, the smaller ThV(os) miss the threshold and SA node becomes quiescent. On reducing high [K(+)](o), ThV(os) re appear, increase in size and initiate spontaneous discharge. As they occur progressively earlier during DD, ThV(os) eventually fuse with V(os): at that stage, DD appears to continue directly into the upstroke (U-shaped DD) and the oscillations are no longer seen. During recovery in Tyrode solution, size and slope of V(os) and of ThV(os) further increase and cause a faster discharge. When APs assume a subsidiary configuration, their DD (no longer U-shaped) abruptly terminates into the upstroke. In high [K(+)](o), increasing [Ca(2+)](o) or applying a fast drive increase the size and slope of V(os) and of ThV(os), which in turn restore or accelerate discharge. In contrast, low [Ca(2+)](o) abolishes V(os) and ThV(os) and causes SA node arrest. Low [Ni(2+)] (35.5 microM) increases the rate whereas high [Ni(2+)] (0.73 mM) stops the SA node. Ryanodine eliminates V(os) and ThV(os) and markedly slows or stops discharge. Thus, ThV(os) and V(os) are separate voltage oscillations that play an obligatory role in the initiation and maintenance of SA node discharge, V(os) by steepening early DD and ThV(os) by attaining the threshold in the dominant pacemaker range, either by gradually increasing during late DD at slow rates or by fusing with V(os) at fast rates. Both V(os) and ThV(os) are Ca(2+) dependent, but apparently in different ways. PMID- 14519437 TI - Overexpression of calpastatin by gene transfer prevents troponin I degradation and ameliorates contractile dysfunction in rat hearts subjected to ischemia/reperfusion. AB - Calpain is a Ca(2+)-activated neutral protease that supposedly plays a key role in myocardial dysfunction following ischemia/reperfusion, by degrading certain proteins involved in the contraction mechanism. It is possible that overexpression of calpastatin, an endogenous calpain inhibitor, lessens contractile dysfunction in the heart after reperfusion by preventing cardiac troponin I (TnI) degradation. This claim is tested by overexpression of human calpastatin (hCS) in rat hearts ex vivo using an adenovirus vector; the hearts were transplanted heterotopically into the abdomens of recipient rats to allow expression of hCS. On the fourth day after surgery, the hearts were excised and perfused in vitro to study their recovery from 30 min of global ischemia, which was followed by 60 min of reperfusion. The peak recovery of the left ventricular developed pressure (LVDP), and the values of its first derivative (max dP/dt, min dP/dt) in the hCS-overexpressed hearts were 88.9 +/- 4.8%, 90.8 +/- 9.2% and 106.4 +/- 9.8%, respectively; these values were all significantly greater than in the control hearts transfected with LacZ alone (51.4 +/- 6.9%, 52.6 +/- 8.1% and 54.7 +/- 6.6%, P < 0.05). In western blot analysis of ventricular myocardial samples (at 60-min reperfusion) using a monoclonal anti-TnI antibody, two bands corresponding to intact TnI (30 kDa) and TnI fragments (27 kDa) were distinguished. The fraction of 27-kDa TnI (percent of total TnI immunoreactivity) in hCS-overexpressed hearts was significantly less than the controls (5.7 +/- 2.7% vs. 18.1 +/- 3.2%, P < 0.05), implying a protective action of hCS against TnI degradation. These results suggest that adenovirus-mediated overexpression of hCS in the heart could be a novel biological means to minimize myocardial stunning by ischemia/reperfusion. PMID- 14519438 TI - Altered signaling surrounding the C-lobe of cardiac troponin C in myofilaments containing an alpha-tropomyosin mutation linked to familial hypertrophic cardiomyopathy. AB - A region of interaction between the near N-terminal of cardiac troponin I (cTnI) and the C-lobe of troponin C (cTnC), where troponin T (cTnT) binds, appears to be critical in regulation of myofilament Ca(2+)-activation. We probed whether functional consequences of modulation of this interface influence the function of tropomyosin (Tm) in thin filament activation. We modified the C-lobe of cTnC directly by addition of the Ca(2+)-sensitizer, EMD 57033, and indirectly by replacing native cTnI with cTnI-containing Glu residues at Ser-43 and Ser-45 (cTnI-S43E/S45E) in myofilaments from hearts of non-transgenic (NTG) and transgenic (TG) mice expressing a point mutation on alpha-Tm (E180G) linked to familial hypertrophic cardiomyopathy. Introduction of cTnI-S43E/S45E induced a significantly greater reduction in tension in TG myofilaments compared to NTG controls. Furthermore, the effect of EMD 57033 to restore Ca(2+)-sensitivity was higher in TG compared to NTG fiber bundles containing cTnI-S43E/S45E and compared to TG or NTG fiber bundles containing native TnI. Our results indicate that alterations in regions of interaction among the N-terminal of cTnI, the C-lobe of cTnC, and the C-terminus of cTnT are important in the regulation of myofilament activity. Although levels of phosphorylation at protein kinase C-dependent sites were the same in TG and NTG myofilaments, our data indicate that the effects of phosphorylation were more depressive in TG hearts. PMID- 14519439 TI - Protective effects of the urocortin homologues stresscopin (SCP) and stresscopin related peptide (SRP) against hypoxia/reoxygenation injury in rat neonatal cardiomyocytes. AB - Urocortin (UCN), a member of the Corticotropin-Releasing Factor (CRF) family of peptides is a well described cardioprotective agent. UCN is able to bind to two types of G-protein coupled receptors: CRF receptor type 1 (CRFR1) and CRF receptor type 2 (CRFR2), whereas, two homologues of UCN, stresscopin (SCP) or also known as urocortin III (UCNIII) and stresscopin related peptide (SRP), or urocortin II (UCNII), bind exclusively and with high affinity to CRFR2, we hypothesised that they will exhibit more pronounced cardioprotective effects than UCN. We show for the first time that SCP is expressed in rat cardiomyocytes and that the levels of SRP and SCP are increased by hypoxic stress. All three peptides have potent cardioprotective effects in cells exposed to hypoxia/reoxygenation. When used at 10(-8) M they increased the amount of live cells by 25% when added prior to hypoxia, and by 20% when UCN and SCP were added at the onset of reoxygenation. In addition, the peptides are equally are more potent antiapoptotic factors than UCN. The antiapoptotic effects of SCP were more pronounced than SRP and UCN at a concentration of 10(-10) M. Furthermore, SCP and SRP protect cardiomyocytes better than UCN at concentrations up to and including 10(-10) M and reduced the amount of TUNEL positive cells almost by half at concentrations of 10(-12) to 10(-10) M. More importantly, we demonstrate that SCP and SRP are able to protect cardiomyocytes even if they are administered after the hypoxic insult and prior to reoxygenation. In this case SCP was more potent than UCN and SRP at 10(-12) M and both SCP and SRP exhibited higher protection at 10(-8) M compared to UCN. Cardioprotection of cardiomyocytes by 10(-8) M of peptides was abolished when treated with 50 microM LY294002 or 100 microM PD98059, but not by 10 microM SB203580 prior to the hypoxic insult. Transfection of dominant negative Akt and MEK1 also blocked protection by the peptides, whereas dominant negative MEKK6 had no effects, demonstrating that SCP and SRP, like UCN, require activation of p42/44 Mitogen activated protein kinase and Akt/Protein Kinase B in order to produce their cardioprotective effects. In addition, we showed that SCP and UCN are potent activators of the p42/44 MAPK pathway, with SRP able to induce phosphorylation of p42/44 MAPK as well, albeit not as pronounced. PMID- 14519440 TI - Effect of muscle origin and phenotype on satellite cell muscle-specific gene expression. AB - Satellite cells from adult mouse tongue, diaphragm, vastus lateralis, rectus femoris, tibialis anterior, soleus, and extensor digitorum longus muscles were isolated, expanded, and differentiated under identical culture conditions. Proliferating myoblasts and differentiated myofiber cultures were analyzed via SDS-PAGE, immunochemical, and PCR methods for expression of myosin heavy chains (MyHC) and muscle creatine kinase (MCK) as indices of muscle fiber type. Contralateral muscles were harvested for simultaneous, parallel analysis utilizing these assays. The MyHC profile of differentiated primary satellite cells was equivalent across all cultures with MyHC(2A) and MyHC(1/slow) co expressed in all myotube and myofiber structures. Trace amounts of MyHC(2B) and MyHC(neo) were detected in a few myofibers. MCK was expressed at a uniform, similar level among these cultures. In contrast, contralateral muscles expressed each muscle-specific indicator at levels correlated with the fiber-type distribution within each muscle. MM14 and C2C12 cells, mouse satellite cell lines, were expanded and compared to primary cell cultures. MM14 cells had a high differentiation index (>95%) and co-expressed MyHC(2A) and MyHC(1/slow) along with trace amounts of MyHC(neo) throughout myotube cultures. Myofibers obtained from C2C12 cells exhibited less differentiation (~75%) with MyHC(2A) as the dominant isoform. These data indicate that primary satellite cells from adult muscle form a uniform differentiated cell type regardless of the fiber-type, anatomic location, and embryonic origin of the donor muscles. MM14 cells expressed an adult MyHC isoform profile similar to primary satellite cells. The results suggest that satellite cells provide a uniform cell source for use in autologous transplantation studies and do not acquire a heritable fiber-type specific phenotype from their host muscle. PMID- 14519441 TI - Sensitization of adenylate cyclase: a general mechanism of neuroadaptation to persistent activation of Galpha(i/o)-coupled receptors? AB - Acute activation of Galphas-coupled receptors stimulates cyclic AMP accumulation leading to the activation of downstream signaling cascades. These Galphas mediated events can be countered by acute activation of inhibitory G proteins (Galpha(i/o)), which inhibit the activity of adenylate cyclase, thereby attenuating cyclic AMP accumulation. Furthermore, an additional, less direct mechanism for Galpha(i/o) proteins modulation of cyclic AMP signaling also has been described. Persistent activation of several Galpha(i/o)-coupled receptors has been shown to result in a subsequent paradoxical enhancement of adenylate cyclase activity in response to drug-stimulated cyclic AMP accumulation. This sensitization of adenylate cyclase likely represents a cellular adaptive response following prolonged activation of inhibitory receptors. Recent advances in our knowledge of G protein signaling, adenylate cyclase regulation, and other cellular signaling mechanisms have extensively increased our insight into this phenomenon. It is now thought that sensitization occurs as part of a compensatory mechanism by which the cell adapts to chronic inhibitory input. Such a mechanism may be involved in modulating Galphas-coupled receptor signaling following neurotransmitter elevations that occur in psychiatric disease states or following the administration of many drugs of abuse. This review will focus on recent advances in the understanding of molecular signaling pathways that are involved in sensitization and describe the potential role of sensitization in neuronal cell function. PMID- 14519442 TI - Gemfibrozil increases the specific binding of rat-cortex nuclear extracts to a PPRE probe. AB - PPAR agonists have been shown to elicit beneficial responses in several cell- and tissue-models of neurotoxicity. To determine if brain PPARs are responsive to the in vivo administration of PPAR agonists in a similar way to those receptors present in other anatomical localizations, such as liver, we fed rats with gemfibrozil incorporated in the diet at a dose that activates hepatic PPARalpha and produces its typical hypolipidemic effect. Rat cortex nuclear extracts presented a pattern of two specific shifted bands when incubated with a PPRE oligonucleotide. Samples from gemfibrozil-treated rats showed a significant increase in the intensity of the two shifted bands regarding control values (2.4- and 1.8-fold for the specific bands 1 and 2, respectively), indicating that orally administered gemfibrozil reaches brain tissues at concentrations sufficient to increase the specific binding of cortex nuclear extracts to an oligonucleotide mimicking a bona fide PPRE, although no changes in cortex ACO mRNA levels were produced. PMID- 14519443 TI - Effect of arachidonic acid on proliferation, cytokines production and pleiotropic genes expression in Jurkat cells--a comparison with oleic acid. AB - The effects of arachidonic (AA) and oleic acids (OA) on proliferation, cytokine production and pleiotropic genes expression in Jurkat T cells were investigated. The following parameters were evaluated: cytotoxicity assessed by loss of membrane integrity and DNA fragmentation, cell proliferation as measured by [14C] thymidine incorporation, production of IL-2, IL-4, IL-10, and INF-gamma, and expression of pleiotropic genes as determined by macroarray technique (83 genes in total). AA was more toxic for Jurkat cells than OA. However, the inhibiting effect of OA on Jurkat cells proliferation was more pronounced than that of AA. The reduction in the production of IL-2 and INF-gamma was more intense by OA (50 microM) than by AA (5 microM). The percentage of genes changed by the fatty acids was: 20.5% (17 genes) for AA (5 microM) and only 2.4% (2 genes) for OA (50 microM). AA markedly affected the expression of genes clustered as: signal transduction pathways, transcription factors and related genes, cell cycle, defense and repair, apoptosis, DNA synthesis, cell adhesion, cytoskeleton and related genes. In particular, AA induced marked changes in cell cycle, signal transduction, and anti-apoptosis genes expression. Therefore, the effect of AA on T-lymphocyte function does involve regulation of expression of important genes, whereas oleic acid did not markedly affect gene expression of Jurkat cells. PMID- 14519444 TI - Effect of Brand's glucosamine with essence of chicken on collagen-induced arthritis in rats. AB - The anti-arthritic effects of glucosamine incorporated in a chicken-meat extract known as Brand's Glucosamine with Essence of Chicken versus glucosamine or Essence of Chicken (EOC) alone were investigated on collagen induced arthritis (CIA) in dark agouti (DA) rats. Four groups of rats received basic food (control), 1.2% glucosamine (GLU), 0.8% EOC and 1.2% GLU + 0.8% EOC (GLU + EOC) admixed with basic food for 25 days following CIA. Foot pads were isolated on day 25 for histopathological evaluation. Clinical assessment of hind paw swelling as measured by foot pad volumes and histopathological scoring based on the degree of edema, periosteal new bone formation, periostitis and inflammatory cell infiltration of the isolated foot pad were performed. Arthritic rats given GLU + EOC showed significant reduction in left hind paw swelling following onset of arthritis. Correspondingly, a lesser degree of edema, periosteal new bone formation, periostitis and inflammatory cell infiltration was seen in histological sections of the left hind foot pads of these rats. A similar trend of reduced hind paw swelling was observed in the right hind paws of the same rats and those fed with EOC. Rats fed with GLU alone did not demonstrate these beneficial effects. The present findings demonstrate that a combination of glucosamine and EOC is effective in reducing the histopathological severity of arthritis, probably due to its ability to reduce the inflammatory conditions in CIA. PMID- 14519445 TI - Study of antihypertensive mechanism of Tribulus terrestris in 2K1C hypertensive rats: role of tissue ACE activity. AB - Tribulus terrestris is a natural herb used for treating many diseases including hypertension. According to previous reports, aqueous extract of tribulus fruits may have some antihypertensive effect with an unknown mechanism. The present study investigated the antihypertensive mechanism of tribulus in 2K1C hypertensive rats by measurement of circulatory and local ACE activity in aorta, heart, kidney and lung. Four groups of rats were selected; control, sham, operated or hypertensive and tribulus treated hypertensive group. Hypertension was induced using silver clip on renal artery by surgery. Four weeks after surgery, a single daily dose of 10 mg/kg of lyophilized aqueous extract of tribulus fruit were given orally to 2K1C rats for four weeks. ACE activity was determined by high performance liquid chromatography (HPLC). Blood pressure was measured by the tail-cuff method. The systolic blood pressure (SBP) was significantly increased in 2K1C rats compared to control rats. The SBP of tribulus fed hypertensive rats was significantly decreased compared to hypertensive rats. The ACE activity in all tissues of 2K1C rats including: aorta, heart, kidney, lung as well as serum were significantly increased compared to normal rats. The ACE activity in all tissues of tribulus fed hypertensive rats was significantly lower than that of hypertensive rats, which was more pronounced in kidney. These results indicated that there is a negative correlation between consumption of tribulus and ACE activity in serum and different tissues in 2K1C rats. PMID- 14519446 TI - Vascular ECE-1 mRNA expression decreases in response to estrogens. AB - DNA microarrays were used to identify new targets of estrogen in the vasculature. Ovariectomized rats were treated with estradiol, genistein or daidzein, for four days. [33P]dCTP-labelled probes synthesized from mesenteric artery RNA were hybridized with DNA microarrays. Analysis of the microarray data identified endothelin converting enzyme-1 (ECE-1) as a gene whose expression was inhibited by treatment with estrogen, genistein, or daidzein. Semi-quantitative RT-PCR was used to confirm the data from the DNA microarrays. Reversal of the estrogen and phytoestrogen effect on ECE-1 expression by ICI 182,780 suggested that the inhibition was an estrogen receptor response. An inhibition of ECE-1 mRNA expression by estrogen or the phytoestrogens has not been previously reported. These data highlight the power of DNA microarray technology to identify new gene expression targets of estrogen in the vasculature. Moreover, the data suggest that genistein and daidzein inhibit ECE-1 expression by an estrogen receptor mediated mechanism. PMID- 14519447 TI - Urease inhibitory activity of simple alpha,beta-unsaturated ketones. AB - A variety of alpha,beta-unsaturated ketones were evaluated for their effect on the jack bean urease. Of 35 compounds tested, 2-cyclohepten-1-one (1), 2 cyclohexen-1-one (2), 2-cyclopenten-1-one (3), and 5,6-dihydro-2H-pyran-2-one (4) showed potent inhibitory activities against the enzyme. The most potent compound (1) (IC50=0.16 mM) showed similar inhibitory potency to hydroxyurea (IC50=0.095 mM). The inhibitory effects of 1, 2, 3, and 4 were significantly reduced by 2 mercaptoethanol or dithiothreitol. These data suggest that alpha,beta-unsaturated ketones inhibited the urease activity, possibly by a Michael-like addition of a protein SH group to the double bond of the alpha,beta-unsaturated carbonyl group. PMID- 14519448 TI - Direct and biochemical interaction between dopamine D3 receptor and elongation factor-1Bbetagamma. AB - Novel signaling components of dopamine D3 receptor (D3R) were searched using yeast two-hybrid system, and the gamma subunit of elongation Factor-1B (eEF1Bgamma) was found to interact with D3R. This interaction was observed specifically between eEF1Bgamma and D3R but not with D2R or D4R. Immunocytochemical studies showed that D3R and eEF1Bgamma form clusters on the plasma membrane and their co-localization was evident in these clusters. The beta subunit of eEF1B (eEF1Bbeta), which forms a tight complex with eEF1Bgamma, was phosphorylated on serine residues in response to the stimulation of D3R. Phosphorylation of eEF1Bbeta was insensitive to pertussis toxin or wortmannin, however, stimulation of cellular protein kinase C (PKC) directly phosphorylated eEF1Bbeta and depletion of PKC abolished D3R-mediated phosphorylation of eEF1Bbeta. These results suggest the involvement of PKC, but not Gi/o proteins or phosphatidylinositol 3-kinase, in D3R-mediated phosphorylation of eEF1Bbeta. Stimulation of D3R did not activate PKC, but the activation of PKC resulted in the phosphorylation of D3R. These results show that PKC has a permissive role for the D3R-mediated phosphorylation of eEF1Bbeta, and suggest that PKC could modulate the mutual interaction between two protein by phosphorylating both D3R and eEF1Bbeta. Therefore, the cellular PKC level would be important for the D3R mediated modulation of eEF1B, and for their cellular regulations such as protein synthesis or cellular proliferation. PMID- 14519449 TI - Structural determinants of blockade of eosinophil activation, adhesion and secretion by synthetic analogs of phomactin. AB - We examined the structural determinants of phomactin analogs to assess their efficacy as antagonist of PAF. Six analogs of phomactin were synthesized to determine their inhibitory effects on adhesion, superoxide release, leukotriene C4 (LTC4) synthesis and [3H]PAF binding in human eosinophils. Phomactin analogs inhibited both PAF- and IL-5-induced eosinophil adhesion. Analog A, which bears an alkene moiety between C-1 and C-14, a ketone at the C-2 position, and an alkyne moiety between C-3 and C-4, had the greatest anti-adhesive effect. Change of the alkene between C-1 and C-14 to an alkane (analog I) decreased the anti adhesive effect by 2.5-4 fold, while substitution of ketone by hydroxyl (analog G) at the C-2 position caused an 11-fold decrease in the anti-adhesive effect. Substitution of the alkyne moiety between C-3 and C-4 by an alkene (B and E) or alkane (D) blocked completely the anti-adhesive effect. Analogs A and I completely blocked superoxide release from eosinophils caused by phorbol-12 myristate-13-acetate or PAF and LTC4-release caused by fMLP plus cytochalasin B. Change of the alkyne moiety between C-3 and C-4 to an alkene (B and E) or alkane (D) blocked completely these inhibitory effects of phomactin. Analog A decreased the maximal binding of [3H]PAF binding to eosinophils without change of the apparent dissociation constant. We conclude that phomactin analogs are specific non-competitive PAF antagonists and have exceptional efficacy in inhibiting adhesion, metabolic activity and leukotriene secretion in human eosinophils. We further define the structural alterations in the phomactin molecule that regulate its inhibitory functions. PMID- 14519450 TI - Supplementation of L-arginine improves hypertension and lipid metabolism but not insulin resistance in diabetic rats. AB - Nitric oxide (NO) plays an important role in glucose and lipid metabolism. We previously reported that NO synthesis inhibitors, such as NG-nitro-L-arginine methyl ester (L-NAME), deteriorate insulin sensitivity and lipid metabolism, while the addition of L-arginine reverses this deterioration. L-arginine is a precursor of NO, and is used as a supplement in the US. In the present study, we evaluated whether the administration of L-arginine alone improves insulin resistance and serum lipid levels in insulin-resistant and hypertriglycemic rat models. Diabetic rats were divided into 3 groups: the control (Cont) group (standard diet), the L-NAME group (diet containing L-NAME), and the Arg group (diet containing L-arginine). After 4 weeks of breeding, urinary NOx, glucose infusion rate (GIR), glucose and lipid tolerance tests were performed. Urinary NOx levels were significantly lower in the L-NAME group than in the Cont group. The GIR in the L-NAME group was significantly lower than that in the Cont group, suggesting increased insulin resistance. However, the administration of L arginine did not influence insulin resistance in the Arg group. Oral lipid administration significantly increased plasma triglyceride levels in the L-NAME group and plasma triglyceride levels were significantly lower in the Arg group than in the Cont group. The area under the curve of plasma triglyceride levels after oral lipid administration was larger in the L-NAME group than in the Cont group. The administration of L-NAME increased insulin resistance and decreased lipid metabolism. L-arginine significantly increased urinary NO secretion but did not improve insulin resistance, although it did improve lipid metabolism. These findings suggest that supplementation of L-arginine cannot improve insulin resistance in diabetic rats probably due to increased insulin secretion by L arginine. PMID- 14519451 TI - Protective effect of SnCl2 on K2Cr2O7-induced nephrotoxicity in rats: the indispensability of HO-1 preinduction and lack of association with some antioxidant enzymes. AB - We have shown that the ameliorative effect of stannous chloride (SnCl2) pretreatment on potassium dichromate (K2Cr2O7)-induced renal damage 24 h after K2Cr2O7 injection was associated with the induction of heme oxygenase-1 (HO-1). In this work we evaluated: (a) if the protective effect of SnCl2 (given 12 h before K2Cr2O7) is associated with changes in the renal activity of HO-1, superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), and catalase (CAT) 24 and 48 h after K2Cr2O7 injection, and (b) if HO-1 induction is indispensable before K2Cr2O7 injection. It was found that the protective effect of SnCl2 on renal function was observed both at 24 and 48 h reaching its maximum at 24 h when HO-1 expression was higher. Cu,Zn-SOD, Mn-SOD, and GR activities remained unchanged whereas GPx and CAT activities decreased at 48 h in K2Cr2O7-treated rats. The activity of Cu,Zn-SOD, Mn-SOD, GPx, CAT, and GR was unchanged in the SnCl2-treated rats. To fulfill the objective (b) groups of rats treated with K2Cr2O7 and SnCl2 (given at the same time or 12 h after K2Cr2O7) were studied 24 h after K2Cr2O7-injection. The simultaneous injections of SnCl2 and K2Cr2O7 had no protective effect whereas the injection of SnCl2 12 h after K2Cr2O7 exacerbated renal damage. In conclusion, the protective effect of SnCl2 on K2Cr2O7-induced nephrotoxicity is associated with HO-1 induction and not with other antioxidant enzymes (Cu,Zn-SOD, Mn-SOD, GPx, GR, and CAT) and SnCl2 has a preventive and not a therapeutic effect on renal damage induced by K2Cr2O7. PMID- 14519452 TI - Brain anatomy and development in autism: review of structural MRI studies. AB - Autism is a neurodevelopmental disorder that severely disrupts social and cognitive functions. MRI is the method of choice for in vivo and non-invasively investigating human brain morphology in children and adolescents. The authors reviewed structural MRI studies that investigated structural brain anatomy and development in autistic patients. All original MRI research papers involving autistic patients, published from 1966 to May 2003, were reviewed in order to elucidate brain anatomy and development of autism and rated for completeness using a 12-item check-list. Increased total brain, parieto-temporal lobe, and cerebellar hemisphere volumes were the most replicated abnormalities in autism. Interestingly, recent findings suggested that the size of amygdala, hippocampus, and corpus callosum may also be abnormal. It is conceivable that abnormalities in neural network involving fronto-temporo-parietal cortex, limbic system, and cerebellum may underlie the pathophysiology of autism, and that such changes could result from abnormal brain development during early life. Nonetheless, available MRI studies were often conflicting and could have been limited by methodological issues. Future MRI investigations should include well characterized groups of autistic and matched healthy individuals, while taking into consideration confounding factors such as IQ, and socioeconomic status. PMID- 14519453 TI - Prenatal morphine exposure suppresses mineralocorticoid receptor-dependent basal synaptic transmission and synaptic plasticity in the lateral perforant path in adult male rats. AB - The effects of prenatal morphine exposure (E11-18) on mineralocorticoid receptor (MR) modulation of synaptic plasticity were investigated in the lateral perforant path (LPP)-dentate gyrus granule cell synaptic system. Hippocampal slices were prepared from adult, prenatally saline- or morphine-exposed male rats. One hour prior to decapitation, some adult male rats were injected subcutaneously with saline or the MR antagonist, canrenoic acid (50 mg/kg). LPP was stimulated with high-frequency (2x100 Hz/0.5 s) and short-term plasticity (STP) and long-term potentiation (LTP) were evaluated at 5 and 30 min poststimulation, respectively. Prenatally saline-exposed male rats injected with saline 1 h prior to decapitation showed significantly higher levels of baseline, STP, and LTP than prenatally saline-exposed, canrenoic acid-treated males. In contrast, prenatally morphine-exposed male rats regardless of saline or canrenoic acid injection 1 h prior to decapitation were comparable in their baseline, STP, and LTP activities. Thus, the results demonstrate that canrenoic acid decreases the efficacy of the basal synaptic transmission in the LPP as well as suppresses synaptic plasticity in saline-exposed males. However, in adult morphine-exposed male rats, canrenoic acid has no other or further effects than a saline treatment suggesting that prenatal morphine exposure suppresses MR-dependent basal synaptic transmission as well as synaptic plasticity. PMID- 14519454 TI - Serial MRI, functional recovery, and long-term infarct maturation in a non-human primate model of stroke. AB - We have examined the effects of permanent middle cerebral artery occlusion (pMCAO) in marmoset monkeys over 5 months, using behavioural and magnetic resonance imaging (MRI) techniques. Three marmosets were trained on behavioural tests before pMCAO. Shortly after surgery, these marmosets were scanned with T2 weighted (T2W) and diffusion-weighted (DW) MRI. Three, 10 and 20 weeks after surgery, these marmosets were re-tested on the behavioural tasks and had further MRI sessions to monitor lesion development. This was followed by histological analysis. All these marmosets had a persistent contralesional motor deficit and a spatial neglect which resolved over the 20 weeks of testing. Percentage infarct volume assessed by MRI on the day of surgery and at 20 weeks matched the percentage infarct volume measured histologically at 20 weeks. However, the apparent infarct size at 3 weeks was considerably less than that measured by histological analysis or that measured at the other MRI time points. Additional histological analysis of the brains of two further marmosets removed 3 weeks after pMCAO found considerable infiltration by lipid filled macrophages into the ischaemic zone which may have caused an MRI "fogging" effect leading to an apparent reduction in infarct volume. PMID- 14519455 TI - Involvement of spinal neurokinin-1 receptors in the maintenance but not induction of carrageenan-induced thermal hyperalgesia in the rat. AB - The study was undertaken to assess the antihyperalgesic effect of L-732,138, (N acetyl-L-tryptophan-3,5-bistrifluoromethyl benzyl ester), a non-peptide neurokinin-1 (NK1) receptor antagonist in rats when given intrathecally. The peripheral inflammation associated with behavioral hyperalgesia to a thermal stimulus was induced by intraplantar (i.pl.) injection of carrageenan. The thermal hyperalgesia was measured by paw withdrawal latency. Intrathecal (i.t.) injection of L-732,138 (100 nmol) at 3h after carrageenan markedly attenuated the paw withdrawal latency of the inflamed paw, but not that of the non-inflamed paw. L-732,138 (100 nmol, i.t.) given 10 min prior to carrageenan injection had no effect on the carrageenan-induced decrease in paw withdrawal latency to noxious thermal stimulus. The results demonstrate that NK1 receptor is involved in the maintenance but not the induction and development of thermal hyperalgesia evoked by carrageenan. PMID- 14519456 TI - Targeting the dopamine D3 receptor cannot influence continuous reinforcement cocaine self-administration in rats. AB - Recent studies point out the important role of dopamine D3 receptors in drug addiction. Therefore, D3 receptor ligands have been proposed as candidate medications for the treatment of cocaine dependence. The present study was designed to compare several dopamine D3 ligands of various selectivity in an animal model of drug-dependence, the cocaine self-administration paradigm. None of the doses of SB-277011 (5, 20 mg/kg), the most selective dopamine D3 antagonist to date, and the lower dose (12 mg/kg) of the moderately D3 selective antagonist U-99194A could influence the rate of self-administration. At the higher dose (24 mg/kg), U-99194A decreased the lever-pressing for cocaine. Both the dopamine D1 selective SCH-23390 (0.2, 0.1 mg/kg) and the dopamine D2 receptor preferring haloperidol (0.5, 0.2 mg/kg) increased the lever-pressing. Both the most dopamine D3 selective agonist PD-128907 (1.0 mg/kg) and the less selective 7 OH-DPAT (0.1, 0.5 mg/kg, s.c.) caused significant decrease in lever-pressing. At lower dose (0.2 mg/kg) PD-128907 was ineffective. The partial agonist BP-897 (1 mg/kg) evoked slight but significant increase in self-administration, while the lower dose (0.5 mg/kg) was ineffective. In all, in contrast to the dopamine D1 and D2 receptors acute inhibition or stimulation of the D3 receptor do not appear to exert considerable influence on the acute reinforcing effect of cocaine. PMID- 14519457 TI - Subtypes of nociceptive units in the rat temporomandibular joint. AB - Response properties of nociceptors in the rat's temporomandibular joint (TMJ) were investigated using an in vitro TMJ-nerve preparation. Recordings were obtained from 33 nociceptive units that responded to mechanical, chemical, and/or thermal stimuli. According to both characteristics of nociceptors and afferent fibers, nociceptive units in the TMJ area were classified into the following four subtypes: Adelta-high-threshold mechanonociceptor (HTM) (12.1%), Adelta-polymodal nociceptor (POLY) (36.4%), C-HTM (12.1%), and C-POLY (39.4%). The mean mechanical threshold of the Adelta units was significantly lower than that of the C units. Bradykinin increased the discharge of Adelta- and C-POLY units. No significant differences of thermal thresholds between Adelta and C units were found. The percentage of Adelta units was 47.2% and of C units was 52.8%, respectively. In the TMJ area, POLY units were predominant (75.8%), suggesting that inflammatory reactions can easily evoke pain sensation. PMID- 14519458 TI - Steroid 21-hydroxylase expression in cultured rat astrocytes. AB - Over the past decade or so it has become widely recognised that the brain is a significant steroidogenic organ. Many publications have highlighted the ability of the brain to synthesise and interconvert a large number of steroid products including cholesterol, progesterone and testosterone. In this study, in vitro experiments were performed to determine if 21-hydroxylation of steroids is undertaken by rat brain astrocytes in culture. This is a common reaction that occurs in the adrenal gland and other organs in mammals, catalysing the conversion of pregn-4-ene-3,20-dione (progesterone) to 21-hydroxypregn-4-ene-3,20 dione (deoxycorticosterone). Previous reports have indicated that 21 hydroxylation occurs within the rat brain, however, the precise identity of the cells expressing 21-hydroxylase has not yet been determined. Several metabolites, such as 5alpha-pregnan-3alpha-ol-20-one (tetrahydroprogesterone) and 3alpha,21 dihydroxy-5-pregnan-20-one (tetrahydrodeoxycorticosterone) were of particular interest because of their modulatory role in neuronal function, such as their agonist activity at gamma-aminobutyric acid (GABA(A)) receptors. Evidence was obtained for the expression of peripheral 21-hydroxylase enzyme (P450c21) in cultured rat brain astrocytes by a combination of mass spectroscopy and molecular biology techniques. This is a significant finding as expression of 21-hydroxylase within astrocytes may be indicative of a wider role for these cells in modulating neuronal behaviour. PMID- 14519459 TI - Differences in hypothalamic Fos expressions between two heat stress conditions in conscious mice. AB - Hyperthermia and dehydration were important physiological phenomena in heat stress. But, the degrees of these phenomena were changed by heat stress conditions, and the distinction between both phenomena is necessary for investigation of response for individual phenomenon. Heat stress at 34 degrees C for 60 min increased rectal temperature, and heat stress at 38.5 degrees C for 60 min further increased rectal temperature and increased osmolality in mice. We investigated the activated region in hypothalamus, which played a role in thermoregulation, fluid regulation and so on, using immunostaining for Fos protein under these conditions in conscious mice. At 34 degrees C, Fos-positive neurons increased in the median preoptic nucleus, lateral preoptic area and anterior hypothalamic area, which were known to be the thermoregulatory center, and the dorsomedial hypothalamic nucleus, which was known to control eating. At 38.5 degrees C, Fos-positive neurons further increased in the regions mentioned above and appeared in the lateral septal nucleus, medial preoptic area, lateral hypothalamic area and zona incerta, which were thought to be involved in thermoregulation and/or fluid regulation, and the paraventricular hypothalamic nucleus, supraoptic nucleus and supraoptic nucleus in the retrochiasmatic part, which were known to be involved in neuroendocrine effector systems. These results support that the activated regions in hypothalamus differed with heat stress conditions, which induced only hyperthermia and both hyperthermia and dehydration. PMID- 14519460 TI - Increased stress vulnerability after a prefrontal cortex lesion in female rats. AB - Neuroimaging studies in patients suffering from affective disorders have shown decreased volume and reduced regional cerebral blood flow in multiple areas of the prefrontal cortex, including the medial prefrontal cortex and the orbitofrontal cortex. This aberrant brain activity is among other things attributed to chronic stress. Affective disorders occur more often in women than in men. In the current experiment, female mPFC-lesioned and non-lesioned rats were subjected to 3 weeks of chronic unpredictable stress in order to determine the role of the mPFC in dealing with chronic stress, and the consequences of mPFC damage for coping with consecutive stressful events. mPFC damage in female rats intensified the stress-induced activation of the dorsomedial nucleus of the hypothalamus and the paraventricular nucleus of the hypothalamus as measured with Fos expression changes and markedly increased plasma catecholamine levels after 3 weeks of unpredictable stress. Additionally, an mPFC lesion significantly reduced the time of appearance of stress-induced behavioral changes in the open field. Altogether, mPFC dysfunction affects the way female rats react to chronic stress, it not only increased the activation of brain regions involved in neuroendocrine and autonomic responses to stress but it also significantly reduced the time of onset of behavioral changes. PMID- 14519461 TI - Effect of resynchronization with GnRH on day 21 after artificial insemination on pregnancy rate and pregnancy loss in lactating dairy cows. AB - The objectives of the present study were to determine the effects of resynchronization with GnRH on Day 21 after artificial insemination (AI) on pregnancy rate and losses of pregnancy in lactating dairy cows. Holstein cows (n=585) on two dairy farms were assigned to one of two treatments in a randomized complete block design. On Day 21 after a pre-enrollment AI, animals assigned to the resynchronization (RES) group received 100 microg of GnRH i.m., whereas animals in the control (CON) group received no treatment. All animals were examined ultrasonographically on Days 21 and 28 after AI, and blood samples were taken for progesterone measurement on Day 21. Pregnancy was diagnosed on Day 28 and reconfirmed 14 days later. Nonpregnant cows on Day 28 were inseminated using timed AI after the completion of the Ovsynch protocol 10 and 17 days after enrollment in the study for RES and CON groups, respectively. Progesterone concentration > or =2.35 ng/ml was used as an indicator of pregnancy on Day 21. For RES and CON cows, pregnancy rate at Days 21 (70.9% versus 73.0%, P<0.56), 28 (33.1% versus 33.6%; P<0.80) and 42 (27.0% versus 26.8%; P<0.98) after the pre enrollment AI did not differ. Administration of GnRH on Day 21 after AI had no effect on pregnancy loss in RES and CON groups from days 21 to 28 (53.2% versus 53.5%; P<0.94) and days 28 to 42 (17.9%; P<0.74) after AI. Pregnancy rate after the resynchronization period was similar for both treatment groups. Resynchronization with GnRH given on Day 21 after AI for initiation of a timed AI protocol prior to pregnancy diagnosis does not affect pregnancy rate and pregnancy loss in lactating dairy cows. PMID- 14519462 TI - Four subpopulations of boar spermatozoa defined according to their response to the short hypoosmotic swelling test and acrosome status during incubation at 37 degrees C. AB - This study was designed to confirm the previously observed relationship between response to the short hypoosmotic swelling test (sHOST) and acrosome resistance in boar spermatozoa. Ejaculates from 22 boars were incubated in a water bath at 37 degrees C for 2h. During the incubation period, samples were taken at 5, 20, 40, 60, 90 and 120 min and subjected to the sHOST. sHOST responses (positive HP negative HN) and acrosomal status (normal or intact NA-damaged DA) were evaluated in 100 spermatozoa corresponding to each ejaculate and incubation time, and the results used to establish four subpopulations: HPNA, HPDA, HNNA and HNDA. Over the entire incubation period, the sHOST positive subpopulation with damaged acrosomes, HPDA, was significantly smaller than the sHOST negative, damaged acrosome subpopulation, HNDA (P<0.001). Further, proportions of HPDA spermatozoa remained stable throughout this period while the HNDA subpopulation showed a significant increase (P<0.001) from the start to the end of incubation. These results confirm the high resistance of the plasma membrane of HP spermatozoa allowing the persistence of a higher number of intact acrosomes over time, compared to HN spermatozoa. Characterising this HPNA subpopulation may help the evaluation of ejaculate quality. PMID- 14519463 TI - Hatching of common carp (Cyprinus carpio L.) embryos stored at 4 and -2 degrees C in different concentrations of methanol and sucrose. AB - The hatching performance of common carp (Cyprinus carpio L.) embryos was examined after 12-72-h storage at 4 and -2 degrees C using different concentrations of sucrose (0.1, 0.25, 0.5 and 1.0 M or 3.42, 8.55, 17.10 and 34.2%), methanol (MeOH) (0.5, 1.0, 1.5, 2.0, 2.5, 3.0 and 3.5 M or 1.6, 3.2, 4.8, 6.4, 8.0, 9.6 and 11.2%), or varying concentrations of methanol in 0.5 M (17.10%) sucrose. For sucrose, 0.5 M (17.10%) showed the maximum survival (41+/-1% (12 h) to 11+/-1.5% (72 h)) at 4 degrees C. No survival was observed at -2 degrees C with any concentration of sucrose. At both temperatures employed, hatching was higher with mixed combination of methanol (1.5 M or 4.8%) and 0.5 M (17.10%) sucrose (4 degrees C: 41+/-1.5% (12 h), 38+/-1.2% (72 h); -2 degrees C: 33+/-1.7% (12 h), 28+/-1.2% (72 h)) compared to methanol alone (4 degrees C: 38+/-1.5% (12 h), 35+/ 2.5% (72 h); -2 degrees C: 31+/-2.5% (12 h), 25+/-2% (72 h)). The combination of 1.5 M (4.8%) methanol and 0.5 M (17.10%) sucrose produced the best results among all the concentrations tested at both temperatures. PMID- 14519464 TI - Suppression of circulating concentrations of FSH and LH by inhibin and estradiol during the initiation of follicle deviation in mares. AB - The role of estradiol and inhibin in suppression of FSH and LH during the initiation of follicle deviation was examined in mares. In Experiment 1, the two largest follicles (F1, F2) were retained during a wave and the rest were ablated as they reached > or =10 mm. The largest follicle was left intact (control, n=12) or was ablated when it reached > or =20.0 mm (Day 0; expected beginning of deviation). The second largest follicle continued growing (n=9) or regressed (n=4) after F1 ablation. Circulating estradiol and total inhibin decreased after Day 0 in the F2-regressing group, whereas estradiol increased after Day 0.5 and inhibin was unaltered in the control and F2-growing groups. Circulating FSH decreased in the control group and increased in the F2-regressing group after Day 0. In the F2-growing group, FSH increased between Days 0 and 0.5 and then decreased. Circulating LH increased between Days 0 and 2 in the F2-regressing group and between Days 0 and 0.5 in the F2-growing group. In Experiment 2, 0 or 1 follicle was retained in a wave followed by administration of 0 or 1 mg of estradiol at the expected beginning of deviation (Hour 0; 2 x 2 factorial design, n=4-6/group). Circulating total inhibin was higher and FSH was lower at Hour 0 in the 1-follicle than in the 0-follicle groups. Follicle-stimulating hormone decreased after Hour 0 in the 1-mg but not in the 0-mg groups, and the decrease in the 0-follicle/1-mg group was not to the level of that in the 1-follicle/1-mg group. Circulating LH was not affected by follicle number but was reduced by estradiol. Results supported the hypotheses that F1 near the beginning of deviation produces inhibin and estradiol and that the increase in circulating estradiol at the beginning of deviation induces FSH suppression in combination with other follicle substances (presumably inhibin). Results also indicated that the increase in estradiol induces suppression of LH. PMID- 14519465 TI - Motility characteristics of boar spermatozoa after addition of prostaglandin F2alpha. AB - Addition of prostaglandin F2alpha (PGF2alpha) to extended boar semen has been shown to slightly increase reproductive parameters in sows such as the conception rate and the total number of piglets born alive. The mechanisms by which PGF2alpha affect these parameters have not yet been elucidated, but it is possible that the sperm transport after insemination is increased. This study investigated whether the sperm motility from 20 Pietrain boars improved when PGF2alpha (Dinolytic; 5 mg PGF2alpha/ml) was added to diluted semen. Different amounts of PGF2alpha (0, 0.5, 1 and 2 ml/100 ml) were tested and the motility was evaluated immediately after addition of PGF2alpha, after 30 min, 2 h, and 24 h. Two computer-assisted semen analysis (CASA) systems, namely the Sperm Quality Analyzer (SQA-IIC) and the Hamilton Thorne (HTR Ceros 12.1) were used to assess the motility parameters. With the SQA-IIC, sperm motility index values of the treated groups were only slightly higher (P>0.05) compared to the negative control group. The different motility parameters measured with the HTR Ceros 12.1 were similar between the treatment groups, except for beat cross frequency, which was higher in the control group (1.5-5%; P<0.001). This study documented that the addition of 2.5, 5 or 10 mg PGF2alpha to 100 ml diluted boar sperm does not increase any sperm motility parameter. Further research is necessary to elucidate mechanisms by which PGF2alpha in diluted semen may improve the reproductive performance in swine farms. PMID- 14519466 TI - Effect of the amount of body condition loss from the dry to near calving periods on the subsequent body condition change, occurrence of postpartum diseases, metabolic parameters and reproductive performance in Holstein dairy cows. AB - We investigated the effect of body condition loss from the dry to near calving periods on the subsequent body condition change, the occurrence of postpartum diseases, the serum metabolic parameters total cholesterol, triglyceride, glucose, and urea nitrogen and the number of days to first breeding after calving in Holstein dairy cows. Body condition scoring (using a 5-point scale with quarter-point divisions) was performed on 67 pregnant Holstein dairy cows. Cows were scored once for body condition during the dry period (prepartum day 35 +/- 16), near calving (postpartum day 5 +/- 4), and subsequently at months 1 (+/- day 4), 2 (+/- day 5), 3 (+/- day 5), and 4 (+/- day 4) of lactation. At the same time, blood samples were collected to evaluate serum total cholesterol, triglyceride, glucose and urea nitrogen concentrations. Cows were categorized based on body condition loss from the dry to near calving periods into two groups: the moderate condition loss group (0-0.75 points, n=41), or the marked condition loss group (1.0-1.5 points, n=26). Regular reproductive health examination was conducted by the same investigator once a week. The marked condition loss group lost more body condition (P<0.01) than the moderate condition loss group consistently until month 1 of lactation. The recovery of body condition in the marked condition loss group was more delayed (P<0.01) than in the moderate condition loss group from months 1 to 4 of lactation. The occurrence of metritis and metabolic diseases (including abomasal displacement, milk fever, ketosis) was greater (P<0.01) in the marked condition loss group (62 and 23%) than in the moderate condition loss group (27 and 2%), respectively. The total cholesterol concentration was lower (P<0.05) in the marked condition loss group (167 +/- 5.3 mg/dl) than in the moderate condition loss group (183 +/- 5.8 mg/dl) at month 1 of lactation, however, the cholesterol levels at the other periods were not different (P>0.05) between the two groups. The triglyceride, glucose, and urea nitrogen concentrations were not different (P>0.05) from the dry period to month 4 of lactation between the two groups. The number of days to first breeding after calving was longer (P<0.05) in the marked condition loss group (103 +/- 7.8 day) than in the moderate condition loss group (87 +/- 5.3 day). We conclude that marked body condition loss from the dry to near calving periods results in the increased occurrence of postpartum metabolic and reproductive diseases, decreased serum total cholesterol concentrations at month 1 of lactation and a longer interval to first breeding after calving in Holstein dairy cows, probably due to the more severe energy deficit reflected by unfavorable body condition score change during early lactation. PMID- 14519467 TI - Overnight incubation improves selection of frozen-thawed blastocysts for transfer: preliminary study using supernumerary embryos. AB - A study was undertaken to determine whether the interval between thawing and transfer influences both biological and clinical outcomes of cryopreserved blastocysts, using supernumerary embryos cultured in sequential media. One hundred and seventy-two patients who underwent blastocyst thawing without any exclusion criteria were included in this single center prospective study of blastocyst thawing cycles. Outcome of 338 blastocysts originating from culture of supernumerary embryos in sequential media was analyzed after 4 or 20 h of culture between thawing and transfer. Survival rate, re-expansion and hatching rates for surviving blastocysts, implantation rates (IRs), pregnancy and miscarriage rates were studied. Blastocyst survival was not influenced by the incubation time after thawing; however both re-expansion and hatching rates were increased after 20-h incubation. Moreover, the IR per thawed or transferred blastocyst was increased three-fold after 20-h incubation compared to 4-h incubation. Increasing the interval between thawing and transfer appears to be beneficial in order to better select for transfer frozen-thawed blastocysts. PMID- 14519468 TI - Bovine embryo development following ICSI: effect of activation, sperm capacitation and pre-treatment with dithiothreitol. AB - The development of bovine embryos obtained by intracytoplasmic sperm injection (ICSI) was studied in relation to various treatments applied to the sperm and to the early embryo. We investigated the effect of different activation protocols on ICSI-embryos and the influence of sperm capacitation with heparin and D penicillamine, hypotaurine, and epinephrine (PHE) prior to ICSI. Finally, we studied the effect of dithiothreitol (DTT) pre-treatment of sperm or of injected oocytes. The activation of ICSI-embryos by ionomycin (Io)-cycloheximide (CHX) and sperm pre-treatment with heparin in combination with PHE did not increase the developmental capacity of ICSI-embryos. By contrast, the treatment of injected oocytes with 2 mM DTT resulted in increased cleavage and blastocyst rates in the group of non-activated embryos and in acceleration of blastocyst development in the group of activated embryos. Similarly, pre-treatment of sperm with DTT, followed by ICSI and activation, determined an increase of embryo development on Day 7 although the total number of blastocysts recorded on Day 8 was not different from untreated controls. The transfer of 11 ICSI-blastocysts, produced without activation, in six recipients gave rise to two pregnancies of which one went to term with the birth of an healthy calf. PMID- 14519469 TI - In vitro fertility and motility characteristics of frozen-thawed boar epididymal spermatozoa separated by Percoll. AB - Frozen-thawed epididymal spermatozoa from four boars were separated through a Percoll gradient, and motility characteristics and in vitro fertility were assessed. Percoll-separated spermatozoa had a significantly higher percentage of motile and progressively motile spermatozoa than those that were not separated (P < 0.0001). However, there were no clear differences in other motility parameters between Percoll-separated and un-separated spermatozoa. Furthermore, sperm agglutination was decreased by Percoll separation (P < 0.05). The effects of Percoll separation on in vitro fertility of spermatozoa differed among boars. In addition, there were large differences in fertility between sperm samples in vitro. Sperm samples, which indicate highly motile and progressively motile, did not always show high in vitro fertility. Furthermore, there was no distinct pattern between fertility in vitro and motility parameters. There was no difference in fertility in vitro between Percoll-separated and un-separated spermatozoa from two of the four boars. However, in vitro fertility of Percoll separated spermatozoa was higher than that of un-separated spermatozoa from the other two boars. PMID- 14519470 TI - Bovine herpesvirus-1 associated with single, trypsin-treated embryos was not infective for uterine tubal cells. AB - It has been reported that bovine herpesvirus-1 (BHV-1) remains associated with in vitro-produced (IVP) bovine embryos after exposure to the virus and either washing or trypsin treatment. However, it is not known if the quantity of virus associated with an exposed IVP embryo is likely to infect a recipient cow after transfer. The specific objective of this study was to determine if IVP embryos that were exposed to BHV-1 would infect uterine tubal cells (UTC) in a co-culture system. In vitro-produced Day 7 embryos were exposed to BHV-1 and then washed or trypsin treated according to the IETS guidelines. These embryos were then co cultured individually or in groups with UTC in microdrops of tissue culture medium 199 (TCM 199) supplemented with 10% equine serum. Following co-culture for 48 h, virus isolation was attempted on the embryos and the UTC from each drop. Virus was detected in washed individual embryos, groups of washed embryos, groups of trypsin-treated embryos and the UTC co-cultured with each of these treatments. However, BHV-1 was not detected in the individual, trypsin-treated embryos or the UTC co-cultured with them. It is concluded that trypsin treatment might effectively prevent infection of recipients if individual, Day 7, exposed embryos were transferred into the uterus. PMID- 14519471 TI - Temporal localization of porcine reproductive and respiratory syndrome virus in reproductive tissues of experimentally infected boars. AB - Porcine reproductive and respiratory syndrome virus (PRRSV) has been reported to be shed in the semen of infected boars. To determine whether the reproductive tissues could be a persistent source of virus and the possible origin of PRRSV found in semen of infected boars, 20 PRRSV-seronegative boars were intranasally inoculated with 5 x 10(6) median tissue culture infective doses (TCID50) of PRRSV and necropsied at different times post-inoculation (p.i.) from Day 2 to Day 37 p.i. Blood samples were collected before experimental inoculation, at necropsy and at different times p.i. At necropsy, epididymal semen and reproductive tissues were collected and the presence of the virus determined by virus isolation. The infection of the boars was demonstrated by the isolation of the virus from the sera of all inoculated boars and by seroconversion. PRRSV was detected in serum samples from Day 2 to Day 23 p.i., although the viremic period was largely dependent on the individual response to infection. Viral replication was proven within different reproductive tissues from Day 2 to Day 23 p.i., being most consistently found in the epididymus. In addition, PRRSV was isolated in semen from Day 4 to Day 10 p.i. The correlation of a diminished viremia and the inability to isolate PRRSV from semen or reproductive tissues may be due to one of two possibilities. First, viremia is responsible for most of the virus isolated from reproductive tissues due to the movement of PRRSV-infected cells out of the blood and into the tissues. Second, viremia may initially seed the reproductive tissues with PRRSV, and then the virus is produced into the reproductive tract and shed into semen at low levels. PMID- 14519472 TI - Surgical recovery and successful surgical transfer of conventionally frozen thawed embryos in the farmed European polecat (Mustela putorius). AB - Surgical transfer of in vivo produced conventionally frozen-thawed embryos of farmed European polecat (Mustela putorius) was investigated as a part of an ex situ preservation program which has the long-term aim of developing a genome resource bank for the endangered European mink (Mustela lutreola). Eighteen oestrous yearling European polecat donors were mated once daily on two consecutive days using 13 fertile males. The donors were surgically flushed for embryos 8-9 days after the first mating. The embryo recovery rate was 60% (116 embryos/193 corpora lutea). The embryos were cryopreserved with 1.5 M ethylene glycol in a programmable freezer using a conventional slow freezing protocol. The thawed embryos were surgically transferred either after dilution with 0.5 M sucrose or directly without removal of ethylene glycol. To induce ovulation, eight recipient females were mated once daily on two consecutive days with vasectomized males starting 7 or 8 days before embryo transfer. The recipients received 7-11 embryos each and three recipients delivered a total of nine pups after a gestation length of 44-46 days. The embryo survival rate was 10% (9 pups/93 frozen embryos). This report describes the first successful cryopreservation of embryos in the Mustelidae family resulting in viable offspring. The low embryo survival rate, however, indicates that the freezing thawing protocol needs to be improved. PMID- 14519473 TI - Influence of stage of lactation and milk production on conception rates after timed artificial insemination following Ovsynch. AB - Conception rates after timed artificial insemination (TAI) are of paramount importance for the success of protocols based on synchronization of ovulation. Stage of lactation and milk production level are known factors that influence dairy cow fertility. It was the objective of this study to analyse the effect of stage of lactation and milk production level on conception rates and pregnancy rates by 200 days in milk (DIM) in dairy cows synchronized with the Ovsynch protocol (Day -10, Day -1: 0.1 mg of D-Phe6-gonadorelin, Day -3: 0.5 mg of cloprostenol, Day 0: AI). A total of 1,288 dairy cows were assigned to two groups and classified in three production levels (high, average, low). Cows of all milk production levels in Group 1 (Simultaneous Ovsynch, SO) were synchronized with the Ovsynch protocol simultaneously for TAI between 73 and 81 DIM. In Group 2 cows with average milk production were synchronized at the same time as Group 1, while low producing cows were synchronized 3 weeks earlier and high producing cows were synchronized 3 weeks later than Group 1, respectively. First service conception rates (FSCRs) were lower (P<0.05) in cows synchronized earlier than in cows of the same production level synchronized later (low production: 14.4% (22/153) versus 34.5% (51/148); high production: 28.2% (40/142) versus 41.4% (53/128)). Milk production level had no significant impact on conception rates after TAI in cows synchronized at the same stage of lactation. At 200 DIM fewer cows with high production level were pregnant than cows with average or low production (P<0.05). This effect was independent of the stage of lactation at the initiation of Ovsynch. Endometritis at a postpartum examination did not influence conception rates after TAI. In conclusion, stage of lactation, but not milk production level, has a major influence on conception rates after TAI. Early AI after Ovsynch is less efficient and therefore its return on investment should be evaluated carefully. PMID- 14519475 TI - Effect of technical settings on canine semen motility parameters measured by the Hamilton-Thorne analyzer. AB - Computerized measuring devices are needed to assess canine semen quality objectively both for research and practical purposes. As internal image settings may influence the results considerably, the effect of different technical settings and semen processing on the parameters assessed by the Hamilton-Thorne Ceros 12.1 semen analyzer (HTR Ceros 12.1) was investigated. The frame rate (15, 30 or 60 frames/s) significantly (P<0.05) influenced most of the measured motility characteristics in experiment 1 while no differences in the motility parameters were found using a different sampling duration (0.5 or 1 s, i.e. 30 or 60 frames scanned) in experiment 2. In experiment 3, an increase in sperm velocity (VAP, VSL, VCL), in linearity and in the percentage of motile and rapidly moving spermatozoa was observed with increasing sperm concentrations (25 x 10(6), 50 x 10(6) or 100 x 10(6) ml(-1)). In experiment 4, a clear effect of the diluent used was visible with higher velocity parameters (VAP, VSL, VCL) and higher percentages of motile, progressive and rapid spermatozoa for semen samples diluted in Hepes-TALP or prostatic fluid in comparison with physiological saline or egg-yolk-Tris extender. In experiment 5, significant (P<0.01) and high correlations were found between the conventional dog semen analysis methods and HTR Ceros 12.1 measurements (n=97 semen samples) for the sperm concentration (r=0.91), the motility (r=0.74) and the progressive motility (r=0.84). In experiment 6, the ejaculates from 21 proven, fertile dogs were compared with the ejaculates of a population (N: 11) of young beagles (1.5 years) but no significant differences in HTR Ceros 12.1 measurements were found between the two groups. Based on our results, diluting dog semen samples to 50 x 10(6) ml(-1) with physiological saline solution and scanning 30 frames at a frame rate of 60 frames/s (i.e. a scanning time of 0.5 s), are the set-up parameters proposed to obtain objective and standardized canine semen motility results using the HTR Ceros 12.1. PMID- 14519474 TI - Comparison of methods to evaluate the plasmalemma of bovine sperm and their relationship with in vitro fertilization rate. AB - The objectives of this study were to compare different methods of evaluating sperm plasmalemma and to determine their relationship with in vitro fertilization rate. A single batch of frozen semen from each of eight beef bulls was used for assessment of sperm viability and for in vitro fertilization. Conventional viability tests included sperm morphology, motility, acrosome integrity, and abnormal DNA condensation. Methods for evaluation of the sperm plasmalemma included eosin/nigrosin (EN) and trypan-blue (TB) vital stains, propidium iodide (PI) in combination with carboxyfluorescein diacetate (CFDA) or SYBR-14 (SYBR) fluorescent vital stains, and the hypoosmotic swelling test (HOST). A total of 133-150 oocytes were fertilized in vitro with sperm from each bull and cleavage rates were determined. There were high correlations between the results obtained with vital stains and good to excellent interclass correlation coefficients of agreement, indicating that these stains provide measures of the same sperm attribute, i.e. plasmalemma integrity. However, the proportions of membrane intact sperm identified by EN or TB stains were greater (P<0.0001) than identified by CFDA/PI or SYBR/PI fluorescent stains. The results obtained with the HOST had moderate correlations but poor agreement with the results of the vital stains. The proportion of viable sperm identified by the HOST was lower (P<0.05) than the proportion identified by vital stains, indicating that response to the HOST did not depend only on the integrity of the plasmalemma. Although there were significant differences in fertilization rates and sperm viability among bulls, there was no sharp distinction for the results of sperm viability tests from bulls producing different in vitro fertilization rates. Proportions of normal, motile, acrosome-intact, and HOST-responsive sperm were identified as significant predictors of in vitro fertilizing potential; each of these endpoints explained 12-18% of the variation when evaluated separately (linear regression) and 48% when evaluated collectively (stepwise regression). In conclusion, EN and TB stains overestimated the proportion of plasmalemma-intact sperm compared to PI based fluorescent stains. Vital stains evaluated the morphological integrity of the plasmalemma, whereas the HOST assessed plasmalemma function. In that regard, the HOST was the only plasmalemma evaluation method that significantly contributed to conventional sperm quality tests in predicting in vitro fertilization rate, indicating that the test could be incorporated to the routine of semen analysis. PMID- 14519476 TI - Chromosomal abnormalities in Day-6, in vitro-produced pig embryos. AB - A cytogenetic study was undertaken to quantify, by chromosomal karyotyping, the incidence and type of chromosomal abnormalities present in Day-6 in vitro produced (IVP) porcine embryos. Morphologically normal Day-6 blastocysts (n=318) were fixed and grouped into six classes according to the number of total cells (from < or =20 to 61-70). Of 248 embryos suitable for analysis, 97 (39.1%) displayed chromosomal abnormalities. The abnormalities included haploidy (9.3%), polyploidy (71.1%) and mixoploidy (19.6%). Within polyploid embryos, triploidy and tetraploidy showed the highest incidence (56.5 and 27.5%, respectively); among mixoploid embryos, diploid-triploid embryos (2n/3n) were prevalent (36.8%). Overall, the mean cell number was 34.3 +/- 12.1 and the mitotic index was 8.6 +/- 6.1. Chromosomally abnormal embryos had fewer (P<0.01) total cells compared to normal (2n) embryos (31.8 +/- 1.3 versus 35.9 +/- 1.0). In addition, the incidence of polyploidy decreased as the number of cells increased, while that of mixoploidy did not differ. These data indicate that polyploidy affects a large percentage of IVP porcine embryos capable of developing to blastocysts and the incidence of chromosomal abnormalities is much higher than that reported previously in in vivo embryos in this species. Given the ability of morphologically normal embryos with an abnormal chromosome complement to undergo preimplantation development in vitro, and the inability to identify blastocysts with abnormal karyotype without cytogenetic analysis, careful consideration should be given to factors affecting ploidy of IVP embryos, especially the incidence of polyspermic fertilization, when evaluating criteria of a porcine in vitro embryo production scheme. PMID- 14519477 TI - Plagiarism revisited. PMID- 14519478 TI - Uterine leiomyosarcoma. PMID- 14519479 TI - Fetal growth restriction and consequences for the offspring in animal models. AB - OBJECTIVE: In the present review we discuss rat models in which intra-uterine growth restriction is obtained through pharmacological (streptozotocin), dietary (global food restriction, low protein diet), or surgical (uterine artery ligation) manipulation of the maternal animal. METHODS: A MEDLINE search was performed on rat models of intrauterine growth restriction (IUGR), ie, streptozotocin, food restriction, low protein diet, or uterine artery ligation and pregnancy and fetal programming, long-term effects or adult offspring. RESULTS: We address the impact of the different maternal conditions for the fetal and neonatal development. The rat models we concentrate on were all associated with fetal hypoinsulinemia and intrauterine growth restriction. Both fetus and neonate adapt to the altered perinatal environment. Some of these adaptations may predispose the offspring to the development of insulin resistance, cardiovascular disease, obesity, and even overt diabetes in later life. CONCLUSION: The adaptations of the fetal metabolism to the altered intrauterine environment have consequences for the offspring, persisting into adulthood and into the next generation. PMID- 14519480 TI - Arginine flux and nitric oxide production during human pregnancy and postpartum. AB - OBJECTIVE: To compare second-trimester, third-trimester, and postpartum arginine flux and nitric oxide production using infusions of the stable isotope L [(15)N(2)]-arginine in normal human gestation. METHODS: Kinetic measurements were made in pregnant volunteers with uncomplicated singleton gestations in mid gestation, late gestation, and more than 8 weeks postpartum. A bolus of 4.95 micromol/kg of labeled arginine was administered, followed by an infusion at 4.95 micromol/kg per hour for 6 hours. The isotopic enrichment of plasma arginine and nitrite or nitrate (NO(x)) was determined by gas chromatography, mass spectrometry, or both. We used the Kolmogorov-Smirnov test for normality, repeated-measures analysis of variance, and Newman-Keuls test. P <.05 denoted statistical significance. RESULTS: The rate of turnover of the intravascular NO(x) pool was significantly higher in mid gestation compared with late gestation and almost reached statistical significance when compared with postpartum values (6.2 +/- 0.9 versus 4.3 +/- 0.8 [P <.02] versus 3.7 +/- 2.1% pool/hour; P =.08). Arginine flux was significantly higher in early compared with late gestation and postpartum (107.8 +/- 13.9 versus 72.5 +/- 16.1 versus 82 +/- 8.8 micromol/kg per hour, respectively; P <.01). CONCLUSION: Arginine and nitric oxide production is higher in mid gestation. This suggests a role for nitric oxide in early cardiovascular adaptation in human gestations. PMID- 14519481 TI - The effect of nuchal cord on amniotic fluid and cord blood erythropoietin at delivery. AB - OBJECTIVE: We investigated the effect of a nuchal cord on fetal hypoxia by using amniotic fluid and cord blood erythropoietin as markers of chronic and acute hypoxia, respectively. METHODS: A total of 167 full-term pregnancies without maternal complications or fetal prelabor complications except fetal growth restriction of unknown cause were studied prospectively. Of these, 47 had a nuchal cord at delivery, and 62 had one or more complications during labor and delivery (nonreassuring fetal heart rate pattern, birth weight less than 2500 g, Apgar score at 1 minute less than 7, presence of meconium-stained amniotic fluid, oligohydramnios), and 26 had both nuchal cord and at least one of the intrapartum complications. RESULTS: Erythropoietin levels (mean +/- standard error of the mean) were not significantly different between the nuchal cord group (n = 47) and the no nuchal cord group (n = 120) in either amniotic fluid (19.3 +/- 4.1 mU/mL versus 13.7 +/- 1.1 mU/mL) or cord blood (57.9 +/- 10.3 mU/mL versus 52.1 +/- 4.9 mU/mL). Similarly, in the 62 fetuses with intrapartum complications, there were no significant differences in amniotic fluid (14.3 +/- 2.0 mU/mL versus 18.8 +/- 2.9 mU/mL) or cord blood erythropoietin (66.9 +/- 16.8 mU/mL versus 72.6 +/- 12.6 mU/mL) levels between those with (n = 26) or without a nuchal cord (n = 36). Among the 107 uncomplicated cases, however, amniotic fluid erythropoietin was significantly elevated in the nuchal cord group (25.5 +/- 8.7 mU/mL, n = 21) compared with that in the no nuchal cord group (11.5 +/- 0.9 mU/mL, n = 84) (P <.05), whereas there was no significant between-group difference in cord blood erythropoietin levels between nuchal cord and no nuchal cord groups (46.8 +/- 10.0 mU/mL versus 43.3 +/- 4.1 mU/mL). Tightness of the nuchal cord did not affect amniotic fluid or cord blood erythropoietin concentrations. CONCLUSION: Although nuchal cord may not significantly increase the risk of acute or labor associated fetal hypoxia, it appears to be an independent risk factor of mild, chronic, prelabor fetal hypoxia. PMID- 14519482 TI - Elevated serum concentrations of the angiogenesis inhibitor endostatin in preeclamptic women. AB - OBJECTIVE: Changes in serum levels of angiogenic factors such as vascular endothelial growth factor or placental growth factor were associated with preeclampsia. The aim of our study was to investigate the concentrations of endostatin, a potent endogenous inhibitor of angiogenesis and endothelial or tumor cell growth, in preeclamptic patients. METHODS: Levels of soluble endostatin were determined by enzyme-linked immunosorbent assay and Western blot analysis in sera of nonpregnant women, healthy pregnant women, and patients with different forms of preeclampsia. RESULTS: Statistical analyses of age-matched study groups revealed elevated medians of endostatin concentration for severely (30.5 ng/mL) and mildly (26.05 ng/mL) preeclamptic patients compared with healthy pregnant (17.2 ng/mL) and nonpregnant women (17.2 ng/mL). Western blot analysis confirmed the up-regulation of soluble endostatin molecules (24 kD) in sera of severely preeclamptic patients. CONCLUSION: Elevated concentrations of endostatin could play a role in the pathophysiology of preeclampsia by counteracting the effects of vascular endothelial growth factor. The inhibitor could also be responsible for the observed growth-inhibitory effects of preeclamptic plasma on endothelial cells. PMID- 14519483 TI - Reduction of preeclampsia in multiple pregnancies by a dedicated monitoring protocol. AB - OBJECTIVE: To evaluate the effect of a dedicated monitoring protocol on the incidence of pregnancy-induced hypertension disorders (preeclampsia, HELLP [hemolysis, elevated liver enzymes, low platelets] syndrome, gestational hypertension) during a 5-year period in 417 women with multiple gestations. METHODS: At the Department of Obstetrics and Fetomaternal Medicine, Vienna University, an outpatient care protocol for women with multiple pregnancies was established. Between March 1997 and February 2002, 379 twins and 38 triplets were followed up. Mean visits of dichorial and monochorial twins were nine and 11, respectively. Triplets were asked to at least 12 follow-up visits. RESULTS: Any kind of pregnancy-associated hypertensive disorders necessitating cesarean delivery developed in eight women (seven with twins, one with triplets; 1.76%). Three cases of preeclampsia, three cases of HELLP syndrome, and two cases of pregnancy-induced hypertension were diagnosed. Perinatal outcome of all 17 newborns was excellent without any mortality and only minor morbidity. All mothers left the hospital in a fully recovered condition. CONCLUSION: Because of our results we hypothesize that our monitoring protocol with frequent visits, continuous personal obstetric care, and timed delivery is effective in reduction of pregnancy-induced hypertension in multiple pregnancies. PMID- 14519484 TI - Interleukin-6 is neither necessary nor sufficient for preterm labor in a murine infection model. AB - OBJECTIVE: The aim of this study was to investigate the role of interleukin 6 (IL 6) in a murine model of bacterially induced preterm delivery. METHODS: On day 14.5 of a 19-20-day gestation, female mice underwent one of two interventions. In experiment 1, pregnant right uterine horns were injected at laparotomy with 0.5 20 microg of recombinant human IL-6 (rhIL-6). In experiment 2, IL-6-deficient (IL 6(-/-)) and wild-type control (IL-6(+/+)) mice underwent intrauterine inoculation with 10(5) to 10(8) heat-killed Escherichia coli organisms. Preterm delivery and maternal survival rates were recorded. RESULTS: In experiment 1, doses as high as 20 microg of IL-6 per mouse resulted in up-regulation of acute phase reactants but did not cause preterm delivery or other adverse maternal or fetal effects. In experiment 2, in bacterially inoculated mice, the absence of maternal and fetal IL-6 had no effect on preterm delivery rates. CONCLUSION: IL-6 was neither sufficient nor necessary for preterm delivery in these murine models. PMID- 14519485 TI - Expression and intracellular localization of protein phosphatases 2A and 2B, protein kinase a, A-Kinase anchoring protein (AKAP79), and binding of the regulatory (RII) subunit of protein kinase a to AKAP79 in human myometrium. AB - OBJECTIVE: To determine the expression and intracellular localization of protein phosphatases 2A (PP2A) and 2B (PP2B), protein kinase A (PKA), and A-kinase anchoring protein (AKAP79), and expression of PKA (RII subunit) binding to AKAP79 in human postmenopausal and pregnant myometrium and to correlate their expressions to blood levels of estradiol, progesterone, and oxytocin. METHODS: Myometrial samples were taken from postmenopausal hysterectomy specimens (group 1, n = 5), from pregnant nonlaboring women (group 2, n = 7) and pregnant laboring women (group 3, n = 5) at cesarean. Western immunoblotting, immunohistochemical, and RII overlay assays were performed. Blood samples were assayed for estradiol, progesterone, and oxytocin levels. RESULTS: There were no significant differences in expression of PP2A, PKA, AKAP79, or PKA(RII) binding to AKAP79 between the three groups. Expression of PP2B was significantly greater in the nonlabor group (group 2) compared with groups 1 and 3. Protein phosphatase 2B, PKA, and AKAP79 expressions were localized in myometrial cytoplasm, but PP2A was localized in blood vessel endothelium. There was no significant correlation between the protein expression and the hormone level in the three groups. CONCLUSION: Human postmenopausal and pregnant (nonlabor and labor) myometrium expressed PP2A, PP2B, PKA, AKAP79, and PKA (RII)-AKAP79 binding. Levels of PP2A, PKA, and AKAP79 expression did not appear to be determinants of human myometrial contractility at parturition. Expression of PP2B may play a role in uterine quiescence. No association was found between protein expression and hormone level. PMID- 14519486 TI - Ripening of the cervix with sodium nitroprusside in nonpregnant women. AB - OBJECTIVE: Few clinical trials have examined the effects of nitric oxide donors on the human cervix. We address the question of the ultrastructural and clinical effects of topical NO donor on the nonpregnant human cervix. METHODS: Twenty patients admitted to the hospital for hysterectomy were randomly assigned to receive either placebo gel or 1% nitroprusside-based gel into the cervical canal. After surgery, a small biopsy of the cervical canal was taken and analyzed by electron microscopy. Vital parameters were monitored after application. RESULTS: In the placebo group, the connective tissue was normal. In the experimental group, the collagen fibers were widely spaced, and no longitudinal collagen bundles were visible. Macrophages, leukocytes, and activated fibroblasts in treated tissues demonstrated a proinflammatory reaction. Adverse effects were reported in a minority of subjects. No significant changes in blood pressure or in nitrite and nitrate levels were reported. CONCLUSION: In both fertile and postmenopausal women, cervical sodium nitroprusside induced morphologic changes similar to those reported for the ripening process. PMID- 14519487 TI - MMP-1 and MMP-2 expression in uterine leiomyosarcoma and correlation with different clinicopathologic parameters. AB - OBJECTIVE: Matrix metalloproteinases (MMPs) have been suggested to play an important role in tumor invasion and metastasis because they degrade a wide range of components of the extracellular matrix. In the present study, we analyzed the expression of MMP-1 and MMP-2 proteins in patients with uterine leiomyosarcoma. METHODS: MMP-1 and MMP-2 expression was investigated by immunohistochemistry from paraffin-embedded tissue sections in 21 patients with uterine leiomyosarcoma (LMS). The immunohistochemical findings were correlated with different clinicopathologic characteristics of the patients. RESULTS: MMP-1 was expressed in 86% and MMP-2 was expressed in 48% of uterine LMS. There was a statistically significant positive correlation between vascular space involvement and MMP-2 expression (P =.05) and between age and MMP-2 expression, with patients over 50 years old having significantly more frequent MMP-2-positive tumors than patients younger than 50 years (P =.006). The relationship between MMP-2 expression and tumor stage and recurrence disease did not reach statistical significance. A trend towards prolonged disease-free survival was observed in women with MMP-2 negative LMS compared with patients with MMP-2-positive LMS (P =.09). Furthermore, a univariate analysis revealed that early tumor stage (P =.0001), age at diagnosis less than 50 years (P =.02), and the absence of vascular space involvement (P =.04) were associated with longer overall survival. CONCLUSION: The statistically significant positive correlation between MMP-2 expression and vascular space involvement as well as the prolonged disease-free survival rate in patients with MMP-2 negative uterine LMS suggest that MMP-2 plays an important role in tumor invasion and metastasis. Further clinical studies with larger numbers of cases need to be performed to verify these findings. PMID- 14519488 TI - Differentiation of ganglion cells and amacrine cells in the rat retina: correlation with expression of HuC/D and GAP-43 proteins. AB - In order to understand the development of retinal cells, we have studied the temporal expression of HuC/D protein in embryonic, postnatal and adult rat retina. During development and in the adult retina, practically all cell somata in the ganglion cell layer and the vast majority of conventional amacrine cells in the inner nuclear layer displayed HuC/D immunoreactivity. Most but not all ganglion cells expressed HuC/D at embryonic day 15, suggesting a delay between final mitosis and the initiation of HuC/D expression. Immunoreactivity for HuC/D was also evident in developing but not mature horizontal cells. Combined immunohistochemical visualization of HuC/D protein and the growth-associated protein (GAP-43) showed a distinct localization of GAP-43 in a specific compartment close to the somato-dendritic region of developing HuC/D-positive cell somata. The localization of GAP-43 immunoreactivity to a specific soma compartment became less evident during maturation. Immunoreactivity for HuC/D and GAP-43 was also discernible in horizontal cells at postnatal day 14. In the adult retina, most GAP-43 immunoreactivity was seen in the inner plexiform layer. Detailed analysis showed that HuC/D and GAP-43 expression is restricted to subsets of retinal neurons during development and in the mature retina. Thus, GAP 43 appears to be correlated with initial steps of differentiation and outgrowth of dendritic processes in HuC/D-positive ganglion and amacrine cells. PMID- 14519489 TI - Differential activation of astrocytes and microglia during post-natal development of dopaminergic neuronal death in the weaver mouse. AB - In order to understand the relationship between astrocytes, microglia and injured neurons, we studied the weaver mutant mouse. One of the main characteristics of this mutant is the progressive degeneration of the dopaminergic (DA) nigrostriatal pathway that starts around postnatal day 15 (P15), in the substantia nigra pars compacta (SNpc) and progresses until adult age (P60). In the present paper, we analysed the relationship between astroglial and microglial cells within DA neurons in the nigrostriatal system of homozygous weaver mice, at different postnatal ages corresponding to specific stages of the DA neuronal loss. The activation of astrocytes was found to be an early event in weaver DA denervation, appearing massively at the onset of DA neuronal loss in the SNpc at P15. Astrocytes remained activated in the adult brain even after the slowing down of the neuronal death process. Interestingly, in the ventral tegmental area, where no DA neuronal death could be detected, a profound, permanent astrogliosis was also observed in adult animals. In contrast, an activation of microglial cells was transiently observed in the SNpc but only at the postnatal age when maximal neuronal death was observed (P30). Lastly, in the striatum, where there was a massive loss of DA nerve terminals, neither astrogliosis nor microglial activation was detected. Hence, the reaction of astrocytes and microglial cells to progressive and spontaneous DA neuronal death showed different temporal kinetics, suggesting a different role for these two cell types in the DA neurodegenerative process in the weaver mouse. PMID- 14519490 TI - Propylthiouracil (PTU)-induced hypothyroidism in the developing rat impairs synaptic transmission and plasticity in the dentate gyrus of the adult hippocampus. AB - Reductions in thyroid hormone during critical periods of brain development can have devastating effects on neurological function that are permanent. Neurochemical, molecular and structural alterations in a variety of brain regions have been well documented, but little information is available on the consequences of developmental hypothyroidism on synaptic function. Developing rats were exposed to the thyrotoxicant, propylthiouracil (PTU: 0 or 15 ppm), through the drinking water of pregnant dams beginning on GD18 and extending throughout the lactational period. Male offspring were allowed to mature after termination of PTU exposure at weaning on PND21 and electrophyiological assessments of field potentials in the dentate gyrus were conducted under urethane anesthesia between 2 and 5 months of age. PTU dramatically reduced thyroid hormones on PND21 and produced deficits in body weight that persisted to adulthood. Synaptic transmission was impaired as evidenced by reductions in excitatory postsynaptic potential (EPSP) slope and population spike (PS) amplitudes at a range of stimulus intensities. Long-term potentiation of the EPSP slope was impaired at both modest and strong intensity trains, whereas a paradoxical increase in PS amplitude was observed in PTU-treated animals in response to high intensity trains. These data are the first to describe functional impairments in synaptic transmission and plasticity in situ as a result of PTU treatment and suggest that perturbations in synaptic function may contribute to learning deficits associated with developmental hypothyroidism. PMID- 14519491 TI - Atm and c-Abl cooperate in the response to genotoxic stress during nervous system development. AB - The c-Abl proto-oncogene is a target of the ATM kinase after DNA double strand breaks, although the physiological significance of these signaling events is not clear. Therefore, to delineate the roles of c-Abl and Atm during mouse development we generated mice with combinations of c-Abl and Atm mutant alleles. We found that dual inactivation of Atm and c-Abl usually resulted in midgestational lethality. However, mice with three mutant alleles, c-Abl(-/ )Atm(+/-) or c-Abl(+/-)Atm(-/-), were viable but predisposed to neuro developmental abnormalities after genotoxic insult. Thus, these genetic data link Atm and c-Abl signaling and underscore a significant interrelationship between the two during neural development. PMID- 14519492 TI - Patterns of cerebral inflammatory response in a rabbit model of intrauterine infection-mediated brain lesion. AB - Although the fetal inflammatory response syndrome seems crucial to the association between intrauterine infection and white matter disease in human preterm infants, the underlying mechanisms remain unclear. Using our previously described rabbit model of cerebral cell death in the white matter and hippocampus induced by intrauterine Escherichia coli infection, we investigated inflammatory and astroglial responses in placenta and brain tissues, in correlation with cell death distribution. Brains and placentas were studied 12, 24, or 48 h following intrauterine inoculation of E. coli or saline (groups G12, G24, and G48). Diffuse monocyte-macrophage infiltrates positive for inducible nitric oxide synthase (i NOS) were significantly more marked in G24 and G48 placentas than in controls. In the G48 fetuses with both diffuse cell death and focal periventricular white matter cysts mimicking cystic periventricular leukomalacia, a strong rabbit macrophage and inducible nitric oxide synthase immunostaining was observed at the border of these cystic lesions. In contrast, in the fetuses with only diffuse and significant cell death, no inflammatory or astroglial responses were detected in the white matter or hippocampus. Cell death was accompanied by i-NOS immunostaining in the hippocampus but not the white matter. Hippocampal cells positive for i-NOS usually displayed a neuronal phenotype. In this model, focal white matter cysts are accompanied by a robust inflammatory response, and diffuse cell death, which may mimic the white matter and hippocampal damage seen in very and extremely pre-term infants, occur in the absence of a detectable brain inflammatory response. PMID- 14519493 TI - Differing patterns of neurotrophin-receptor expressing neurons allow distinction of the transient Frorieps' ganglia from normal DRG before morphological differences appear. AB - The Frorieps' ganglia are dorsal root ganglia (DRG) that form and then degenerate during normal embryonic development of amniotes. Their degeneration or survival has been shown to be modulated by modifying expression of Hox-family and other genes involved in pattern formation, and by the mesodermal microenvironment of the cranial somites in which they develop. In ovo application of the neurotrophin NGF partially rescues DRG2 from degeneration. To further examine the potential role of neurotrophins in the life cycle of Frorieps' DRG we have now quantified the numbers of neurons expressing neurotrophin receptors trkA and trkC in avian Frorieps' ganglia (DRG2) and normal cervical DRG (DRG5). We have found that the Frorieps' DRG are different from normal DRG in terms of the numbers of neurons expressing these receptors. trkC-expressing neurons are generally lacking in DRG2, this is the earliest (St 18, E2.5) described difference between DRG2 and normal DRG, preceding morphological differences between these ganglia that appear at St 20. The difference between DRG2 and DRG5 in terms of numbers of trkA expressing neurons is evident only at later embryonic stages, where DRG2 contains a higher proportion of trkA neurons than normal cervical DRG. The few trkC+ neurons present late in DRG2 development are not concentrated in the VL portion of the ganglion, the zone where trkC+ neurons are generally found in normal DRG. We also find that DRG2 neurons are smaller than those of normal DRG, this is true for both trkA+ and trkC+ populations. These data together therefore suggest that the neurons that survive in the Frorieps' ganglia at later stages belong almost exclusively to the trkA-expressing DM class DRG neurons. We further find that the differences in the populations of trkA/trkC between DRG2 and DRG5 result from signals from the mesodermal microenvironment, since DRG arising in cranial somites transplanted caudally contain few trkC+ neurons and a higher proportion of trkA+ cells than contralateral controls. PMID- 14519494 TI - Muscarinic calcium mobilization in the regenerating retina of adult newt. AB - We used optical recording with a Ca(2+)-sensitive dye, fura2, in living slice preparations from the newt retina at different stages of regeneration. ACh produced the most pronounced [Ca2+]i rise in progenitor cells and premature ganglion cells of the earlier stage of retinal regeneration, but less pronounced Ca2+ response in ganglion cells just before, or at the beginning of, synaptogenesis. The [Ca2+]i rise to ACh was mediated by mAChRs. This was shown by inhibition of the ACh-induced Ca2+ response with a preincubation of the mAChR antagonist atropine as well as with direct stimulation of the [Ca2+]i rise by the mAChR agonist muscarine. This muscarine-induced [Ca2+]i rise was more greatly suppressed by the M1 and/or M3 preferring mAChR antagonists than by the M2 preferring mAChR antagonist. The [Ca2+]i rise due to muscarine was not suppressed in the absence of extracellular Ca2+, but suppressed in part in the presence of the L-type voltage-gated Ca2+ channel blockers, verapamil or nicardipine. Furthermore, thapsigargin (TG), a Ca-ATPase inhibitor, abolished the muscarine induced [Ca2+]i rise in the absence of extracellular Ca2+. These results suggest that the mAChR-mediated [Ca2+]i rise is mainly a result of a release of Ca2+ from intracellular stores. TG produced a slow rise in the resting level of [Ca2+]i. This [Ca2+]i raise was suppressed as extracellular Ca2+ was omitted, whereas a rapid rise in [Ca2+]i occurred when extracellular Ca2+ was reintroduced, suggesting the occurrence of the capacitative Ca2+ influx in the progenitor cells and premature ganglion cells of the regenerating newt retina. PMID- 14519495 TI - Effects of gonadal steroids on pineal morphogenesis and cell differentiation of the embryonic quail studied under cell culture conditions. AB - Receptors for gonadal steroid hormones have been localized in the pineal glands of several vertebrate species. No studies, however, have reported on pineal morphogenesis and cell differentiation following hormonal application in vitro during avian embryonic development. Hormonal regulation of embryonic development is crucial in all vertebrate classes. Although gonadal hormones are known to affect organogenesis in avian embryos and chicks, we wanted to investigate whether gonadal steroids (testosterone and estradiol) have any effect on the morphogenesis and cell differentiation of the avian pineal gland. The steroid hormones had a stimulatory influence on pineal morphogenesis in vitro as evidenced from the radial arrangement of colony-forming cells and the subsequent formation of a follicular-like structure under dispersed-cell culture condition. Administration of testosterone in culture medium significantly promoted the numbers of cells that were positively stained for arginine vasopressin and tyrosine hydroxylase, while estradiol showed only a slight effect. Both of the two steroid hormones significantly decreased the numbers of cells positively stained for serotonin and melatonin. Melatonin released in the culture medium decreased in content within the 24 h following steroid treatment (supported by low immunoreactivity in cultured cells and low level released to the medium). These results clearly suggest active roles of gonadal steroid hormones on embryonic pineal morphogenesis and cell differentiation and its physiological activity as they do in adult animals. PMID- 14519496 TI - Intracerebroventricular kainic acid administration to neonatal rats alters interneuron development in the hippocampus. AB - The effects of neonatal exposure to excitotoxins on the development of interneurons have not been well characterized, but may be relevant to the pathogenesis of neuropsychiatric disorders. In this study, the excitotoxin, kainic acid (KA) was administered to rats at postnatal day 7 (P7) by intracerebroventricular (i.c.v.) infusion. At P14, P25, P40 and P60, Nissl staining and immunohistochemical studies with the interneuron markers, glutamic acid decarboxylase (GAD-67), calbindin-D28k (CB) and parvalbumin (PV) were performed in the hippocampus. In control animals, the total number of interneurons, as well as the number of interneurons stained with GAD-67, CB and PV, was nearly constant from P14 through P60. In KA-treated rats, Nissl staining, GAD-67 staining, and CB staining revealed a progressive decline in the overall number of interneurons in the CA1 and CA3 subfields from P14 to P60. In contrast, PV staining in KA-treated rats showed initial decreases in the number of interneurons in the CA1 and CA3 subfields at P14 followed by increases that approached control levels by P60. These results suggest that, in general, early exposure to the excitotoxin KA decreases the number of hippocampal interneurons, but has a more variable effect on the specific population of interneurons labeled by PV. The functional impact of these changes may be relevant to the pathogenesis of neuropsychiatric disorders, such as schizophrenia. PMID- 14519497 TI - Development of the outer retina in the mouse. AB - Mice represent a valuable species for studies of development and disease. With the availability of transgenic models for retinal degeneration in this species, information regarding development and structure of mouse retina has become increasingly important. Of special interest is the differentiation and synaptogenesis of photoreceptors since these cells are predominantly involved in hereditary retinal degenerations. Thus, some of the keys to future clinical management of these retinal diseases may lie in understanding the molecular mechanisms of outer retinal development. In this study, we describe the expression of markers for photoreceptors (recoverin), horizontal cells (calbindin), bipolar cells (protein kinase C; PKC) and cytoskeletal elements pivotal to axonogenesis (beta-tubulin and actin) during perinatal development of mouse retina. Immunocytochemical localization of recoverin, calbindin, PKC and beta-tubulin was monitored in developing mouse retina (embryonic day (E) 18.5 to postnatal day (PN) 14), whereas f-actin was localized by Phalloidin binding. Recoverin immunoreactive cells, presumably the photoreceptors, were observed embryonically (E 18.5) and their number increased until PN 14. Neurite projections from the immunoreactive cells towards the outer plexiform layer (OPL) were noted at PN 0 and these processes reached the OPL at PN 7 coincident with histological evidence for the differentiation of the OPL. Outer segments, all the cell bodies in the ONL, as well as the OPL were immunoreactive to recoverin at PN 14. Calbindin immunoreactive horizontal cells were also present in E 18.5 retinas. These cells became progressively displaced proximally as the ONL developed. A calbindin immunoreactive plexus was seen in the OPL at PN 7. PKC immunoreactive bipolar cells developed postnatally, becoming distinguished at PN 7. Both beta-tubulin and actin immunoreactive cells were present in the IPL as early as E 18.5; however, appearance of processes labeled with these markers in the OPL was delayed until PN 7, concurrent with the first appearance of photoreceptor neurites, development of the horizontal cell plexus, and development of synaptophysin immunoreactivity at this location. These results provide a developmental timeframe for the expression of recoverin, calbindin, synaptophysin, beta-tubulin and actin. Our findings suggest that the time between PN 3 and PN 7 represents a critical period during which elements of the OPL are assembled. PMID- 14519499 TI - Distribution of estrogen receptor alpha and beta immunoreactive profiles in the postnatal rat brain. AB - The present study was conducted to identify the localization and possible contribution of the two estrogen receptor (ER) subtypes in the rat brain at postnatal (P) days 3, 7 and 14. Evaluation of the distribution of ERalpha and ERbeta immunoreactive (ir) nuclei did not reveal gender differences at the developmental point times examined. With the exception of the cerebral cortex, the pattern of staining for these ERs was unchanged across the postnatal ages examined. The distribution of ERalpha-ir nuclei was wider than ERbeta-ir during brain development. From P3, ERbeta and ERalpha-ir nuclei were found in different regions of the cerebral cortex, basal forebrain, amygdala, thalamus, hypothalamus, mesencephalon, pons, cerebellum and medulla oblongata. In addition, ERalpha-ir nuclei were exclusively detected in the hippocampal subfields, epithalamus and in several circumventricular organs. ERalpha and ERbeta dual immunofluorescence revealed positive nuclei in the medial part of the bed nucleus of the stria terminalis, periventricular preoptic nucleus and in caudal aspects of the ventrolateral part of the ventromedial hypothalamic nucleus. Although the functional significance of the dual expression of both ERs within the same nuclei remains unknown, it is possible that ERs play different roles in gene regulation within the same cell. The presence of ERs in diverse brain regions through early postnatal periods supports a potential role for estrogens in neural differentiation. PMID- 14519498 TI - The transcription factor Nurr1 in human NT2 cells and hNT neurons. AB - Human, neuronally committed hNT or NT2-N cells, originally derived from the Ntera2/D1 (NT2) clone after exposure to retinoic acid (RA), represent a potentially important source of cells to treat neurodegenerative diseases. Our previous in vitro experiments showed that hNT cells possess immunocytochemically detectable markers typical of dopaminergic (DA) ventral mesencephalic (VM) neurons, including tyrosine hydroxylase (TH), dopamine transporter (DAT), dopamine receptor (D2), and aldehyde dehydrogenase (AHD-2). In the current study, we sought to examine whether Nurr1, an orphan receptor of the nuclear receptor superfamily shown to be essential for the development, differentiation and survival of midbrain DA neurons, would be expressed in 3, 4, or 5 week RA-induced hNT neurons and their NT2 precursors. Our immunocytochemical analyses indicate that NT2 cells as well as hNT neurons independent of the length of RA-driven differentiation were Nurr1-immunoreactive. RT-PCR analysis confirmed the expression of Nurr1-specific mRNA in both NT2 precursors and the hNT neurons. Furthermore, immunocytochemical co-expression of Nurr1 and TH was detected in hNT neurons. The findings of this study suggest that Nurr1 may be important during the development of hNT neurons and involved in their differentiation into the dopaminergic phenotype. PMID- 14519500 TI - Interactions between human adipose stromal cells and mouse neural stem cells in vitro. AB - Transplantation of adult mesenchymal stem cells (MSCs) into adult rat brain has been known to reduce functional deficits associated with stroke and traumatic brain injury. However, in injured brains, there is no evidence that transplanted MSCs replace lost host brain tissue. In this study, we determined in vitro interaction between human adipose tissue stromal cells (hATSCs), a kind of MSC, and neural stem cells (NSCs). hATSCs were isolated and proliferated from human adipose tissues, and NSCs from the subventricular zone of postnatal mice. When NSCs were cultured on mitomycin-treated hATSC monolayers, their proliferation was decreased, but neuronal differentiation was significantly induced. The percentage of neurons significantly increased in 7 days in cultures of NSCs on hATSCs feeder as compared to NSCs cultured on laminin-coated dishes. When the duration of the cultures was extended to 14 days, hATSCs supported the survival of neurons derived from NSCs. To determine the role of soluble factors from hATSCs, NSCs were cultured with hATSCs conditioned medium or co-cultured with permeable filter on which hATSCs were grown. Although proliferation of NSCs significantly decreased and glial differentiation increased under these experimental conditions, their neuronal differentiation was not affected, indicating that direct physical contact between hATSCs and NSCs is required for induction of neuronal differentiation. These data indicate that hATSCs may provide supportive roles on endogenous neural stem cells, when they are transplanted into damaged brain. PMID- 14519501 TI - Sex differences in estrogen-induced progesterone and estrogen receptor mRNA in the ventromedial hypothalamus of hatchling whiptail lizards. AB - The ventromedial hypothalamus (VMH) is an important neural locus for the control of female-typical sexual behavior in vertebrates, and exogenous estrogen stimulates a strong increase in progesterone receptor (PR) in the VMH of adult females. Estrogen also regulates its own receptor (ER), though the direction of the response varies from species to species. In rodents and whiptail lizards, males either lack estrogen regulation of PR and ER mRNA in the VMH or display a greatly attenuated response. We examined hatchlings of two closely related species of whiptail lizards, one of which is parthenogenetic. Though normally all female, the parthenogens can be made to develop as gonadal males by treating with aromatase inhibitor early in development. Thus, we were able to ask whether the brain sex of these 'created male' parthenogens corresponded to their gonadal sex or their genetic sex. We injected 1- and 30-day-old animals of both species and sexes with estradiol benzoate (EB) and assayed for PR and ER mRNA using in situ hybridization. All animals given EB responded with a strong increase in PR mRNA in the VMH. However, females of the sexual species had higher EB-induced PR mRNA levels than did conspecific males; there was no sex difference between the normal parthenogens and the created males of the parthenogenetic species. EB also stimulated an increase in ER mRNA in the VMH, but the pattern of response was more complex. Normal parthenogens did not increase ER mRNA in response to EB in either age group, in contrast to the strong response of 1-day-old males and females of the sexual species and 30-day-old created males. The results indicate that hatchling whiptails show striking species and sexual differences in the regulation of sex steroid receptor mRNAs in an area of the brain important for adult sexual behavior. This variation may play a role in the development of species and sexual differences in the adult neuroendocrine phenotype. PMID- 14519502 TI - Expression of the BMP antagonist Dan during murine forebrain development. AB - Dan (Differential screening-selected gene aberrative in neuroblastoma, also known as N03) is a member of a class of glycoproteins shown to be secreted inhibitors of the transforming growth factor-beta (TGF-beta) and bone morphogenic protein pathways. We examined Dan expression during murine forebrain development from embryonic day 10.5 to postnatal day 14 and found that Dan was expressed in highly specific spatiotemporal patterns. In early telencephalic development, Dan is highly expressed in the fibroblasts covering the cortex. From E12.5-E14.5, Dan is also weakly expressed in a region of neuroepithelium at the medial margin of the telencephalon called the cortical hem. From E17.5 on, Dan is expressed strongly in CA3 pyramidal neurons of the hippocampus as well as in the developing thalamus and amygdala. To determine if Dan expression is correlated with the expression of any of its known ligand targets, we examined the expression of GDF-5, -6 and -7 in the forebrain and found that GDF-5 is expressed in Cajal-Retzius cells in Layer I of cortex, immediately adjacent to the expression of Dan in the meninges. PMID- 14519503 TI - Oral-motor patterns of rhythmic trigeminal activity generated in fetal rat brainstem in vitro. AB - Development of neural circuits generating fetal oral-motor activity was characterized in an in vitro isolated brainstem block preparation. Rhythmical trigeminal activity (RTA) at E20-E21 resembled either the pattern or rhythm of neonatal RTA. Conversely, at E18-E19, RTA displayed a different pattern of discharge from neonatal RTA, and output was not regular but intermittent with another slow rhythm. PMID- 14519505 TI - The distributions and signaling directions of the cerebrospinal fluid contacting neurons in the parenchyma of a rat brain. AB - Many studies have been made on the distributions of CSF contacting neurons (CSF CNs) in the parenchyma of the brain with horseradish peroxidase (HRP) or autoradiographics. A significant amount of data has shown that both HRP and autoradiographical substances could pass freely through the spaces of ependyma into the parenchyma of the brain. It is therefore possible that the results were not exact. We found that CB-HRP was a dependable tracer to CSF-CNs and studied the distributions and the signaling directions of cerebrospinal fluid contacting neurons (CSF-CNs) in the parenchyma of the brain with the cholera toxin subunit B with horseradish peroxidase (CB-HRP) tracing combined with transmission electron microscopy. The results were as follows: (1) CSF contacting tanycytes existed not only in the wall of the third ventricle (3V), but also in the walls of the lateral ventricle (LV), the fourth ventricle (4V) and the central canal (CC) of the spinal cord. (2) Some CSF contacting glia cells were observed in the lateral septal nucleus (LS). (3)The distal CSF-CNs in the parenchyma were found in LS, the anterodorsal thalamic nucleus (AD), the supramammillary nucleus (SuM), the dorsal raphe nucleus (DR), the floor of 4V and the lateral superior olive (LSO), but they were mainly found in DR and divided into groups A and B. (4) Axon terminals labeled by CB-HRP were found in the cavity of the brain ventricle. (5) The synaptic relationships between the neurons were labeled by CB-HRP in DR and no-labeled by CB-HRP in the parenchyma. Both synapses Gray I and II were found. It was significant that the presynaptic elements were formed by the neurons no labeled CB-HRP and the postsynaptic elements labeled CB-HRP. Our results suggested firstly that the signaling directions of CSF-CNs in DR were only from the parenchyma to CSF. PMID- 14519506 TI - Angiogenic response induced by acellular brain scaffolds grafted onto the chick embryo chorioallantoic membrane. AB - The repair and regeneration of injured tissues and organs depend on the re establishment of the blood flow needed for cellular infiltration and metabolic support. Among the various materials used in tissue reconstruction, acellular scaffolds have recently been utilized. In this study, we investigated the angiogenic response induced by acellular brain scaffolds implanted in vivo onto the chick embryo chorioallantoic membrane (CAM), a useful model for such investigations. The results show that acellular brain scaffolds are able to induce a strong angiogenic response, comparable to that of fibroblast growth factor-2 (FGF-2), a well known angiogenic cytokine. The response may be considered dependent on a direct angiogenic effect exerted by the scaffold, because no inflammatory infiltrate was detectable in CAM's mesenchyme beneath the implant. Acellular brain scaffolds might induce the release of endogenous angiogenic factors, such as FGF-2 and vascular endothelial growth factor (VEGF) released from the extracellular matrix of the developing CAM. In addition, the angiogenic response may depend, in part, also on the presence in the acellular matrix of transforming growth factor beta 1 (TGFbeta1). PMID- 14519507 TI - Evidence for a persistent Na-conductance in identified command neurones of the snail, Helix pomatia. AB - Neurones RPa3 and LPa3 were identified as 'command' neurones in the Helix parietal ganglia. The physiological role of these cells is the integration of sensory information before triggering withdrawal behaviour. Properties of the Na channels are poorly understood in these neurones which produce Na(+)-dependent action potentials in Ca(2+)-free solution. Our aim was to describe the kinetic properties and TTX-sensitivity of the Na-channels of these cells, and to provide evidence for the existence of a persistent inward sodium current (I(NaP)) in them. Two-microelectrode voltage- and patch-clamp techniques were used on isolated or semi-isolated neurones. The kinetics and potential dependence of the transient inward sodium current (I(NaT)) agreed well with those obtained on other molluscan neurones. We concluded that I(NaT) present in these neurones is slow and TTX-resistant (k(D)=8 microM of TTX) and has two components with different rates of inactivation. In addition, the presence of an I(NaP) component was revealed. We showed that I(NaP) is neither an artifact nor the contribution of a Ca-channel or a 'window' current. With slow voltage ramp pulses I(NaP) could be activated and separated from I(NaT). Like I(NaT) it appeared to be TTX-resistant and Na-dependent. I(NaP) was upregulated by increased pH (8.0) and decreased by elevated extracellular Mg(2+) concentration parallel with the I(NaT). Our results suggest that I(NaP) originates from the same set of sodium channels that underlie I(NaT). PMID- 14519508 TI - Long term potentiation varies with layer in rat visual cortex. AB - Long term potentiation (LTP) in various layers of rat visual cortex was studied in 90 cells with visually identified, whole-cell recordings. LTP was induced in layer II/III, layer V or layer VI with theta burst stimulation (TBS), but was not observed in layer IV. In the presence of a NMDA antagonist, D-AP5, in the bath solution, potentiation was blocked in layer II/III, some depression was seen in layer V, and potentiation still remained in layer VI. After addition of a specific mGluR1 antagonist, LY367385, to the bath solution, LTP was reduced in layer II/III and layer V, and was blocked in layer VI. After a specific mGluR5 antagonist, MPEP was applied in the bath solution, LTP was enhanced in layer VI, and blocked in layer V. We conclude that: (1) LTP in layer VI is different from other layers, depending on mGluR1, but not NMDA receptors. (2) In layer II/III, LTP is NMDA-dependent and is not blocked by group I mGluR antagonists. (3) LTP in layer V is both NMDA receptor and mGluR5 receptor-dependent. (4) LTP was not induced in layer IV with TBS. PMID- 14519509 TI - Expression of D4 dopamine receptors in striatonigral and striatopallidal neurons in the rat striatum. AB - Recent studies have reported the regional distribution of D(4) dopamine receptors in the rat striatum at the cellular and subcellular levels. However, the precise identity of the striatal neurons that express these receptors remains unknown. We have studied the expression of D(4) receptors in the striatal interneurons as well as in the output regions of the striatum using immunohistochemistry. Furthermore, we have evaluated the contribution of the striatum to D(4) receptor immunoreactivity in these areas by means of ibotenic acid lesion of the striatum. D(4) receptors were observed in the substantia nigra pars reticulata (SNr), the entopeduncular nucleus (EP) and the globus pallidus (GP), and they were found, using electron microscopy, to be located presynaptically. D(4) immunoreactivity in the striatal output nuclei was observed to dramatically decrease following lesion of the striatum with ibotenic acid. Striatal interneurons were not found to express D(4) receptors. These results demonstrate that D(4) receptors are located almost exclusively in striatal projection neurons, in both striatonigral and striatopallidal neurons. PMID- 14519510 TI - Serotonin 5-HT1A receptors might control the output of cortical glutamatergic neurons in rat cingulate cortex. AB - The present study was designed to investigate the distribution of serotonin 5 HT1A receptor protein (5-HT1A-immunoreactivity) and its localization within cortical pyramidal neurons of the rat cingulate cortex. This experimental direction was inspired by recent data showing the role of 5-HT1A receptors in the pathology of schizophrenia, and in the mechanism of action of novel antipsychotic drugs as well as by the importance of the cingulate cortex in regulation of cognitive functions. It was found that 5-HT1A-immunoreactivity was densely distributed in neuronal eyelash-like elements, and their size, shape and spatial orientation may suggest concentration of 5-HT1A-immunopositive material in the proximal fragments of axons of cortical neurons. Moreover, it was observed that these 5-HT1A-immunopositive fragments were present predominately on proximal fragments of axons of pyramidal neurons, which was evidenced by double labeling experiments using glutamate and non-phosphorylated neurofilament H as markers of the cortical pyramidal cells. The 5-HT1A receptor immunoreactivity was localized distally to the inhibitory GABAergic terminals of chandelier and basket cells surrounding the pyramidal cell bodies and occasionally surrounding short initial segment of axonal hillock of pyramidal neurons. These anatomical data indicate that 5-HT1A receptors might control the excitability and propagation of information transmitted by the pyramidal cells. Moreover, our results indicate that drugs operating via 5-HT1A receptors in the cingulate cortex might control from this level the release of glutamate in the subcortical structures. Finally, the 5-HT1A receptors present in the cingulate cortex, as demonstrated in the present study, may constitute an important target for drugs used to repair dysfunction of glutamate neurotransmission, which is observed for example in schizophrenia. PMID- 14519511 TI - Cholinergic axons in the rat prostate and neurons in the pelvic ganglion. AB - Fluorogold or green fluorescent pseudorabies virus labeled postganglionic neurons in the pelvic ganglion that innervate the prostate gland. Small cholinergic neurons were demonstrated by immunohistochemistry (IHC) with antiserum against vesicular acetylcholine transferase (VAChT). Large, mainly adrenergic neurons, were surrounded by preganglionic cholinergic boutons. In the prostate, M3 type muscarinic receptors were found in the outer muscle layer surrounding the prostatic acini. The antiserum against VAChT marked the inner epithelial layer. Antisera against the vesicular monoamine transporters VMAT1 and VMAT2 demonstrated staining of the inner secretory layer and adrenergic fibers in the outer muscle layer, respectively, of the prostatic acini. These results provide new evidence for the presence of neural elements that have a cholinergic influence over the rat prostate gland. PMID- 14519512 TI - Deleterious poly(ADP-ribose)polymerase-1 pathway activation in traumatic brain injury in rat. AB - Traumatic brain injury produces nitric oxide and reactive oxygen species. Peroxynitrite, resulting from the combination of nitric oxide and superoxide anions, triggers DNA strand breaks, leading to the activation of poly(ADP ribose)polymerase-1. As excessive activation of this enzyme induces cell death, we examined the production of nitrosative stress, the activation of poly(ADP ribose)polymerase-1, and the role of this enzyme in the outcomes of traumatic brain injury produced by fluid percussion in rats. Immunohistochemistry showed that 3-nitrotyrosine, an indicator of nitrosative stress, and poly(ADP-ribose), a marker of poly(ADP-ribose)polymerase-1 activation, were present as early as 30 min post-injury, and that persisted for 72 h. The poly(ADP-ribose)polymerase inhibitor, 3-aminobenzamide, at 10 and 30 mg/kg, significantly improved the neurological deficit, with a 60% reduction in the brain lesion volume and inhibition of poly(ADP-ribose)polymerase-1 activation. Thus, poly(ADP ribose)polymerase-1 is involved in the neurological consequences of traumatic brain injury and may be a promising therapeutic target in clinical treatment of acute brain trauma. PMID- 14519513 TI - Pharmacokinetics and pharmacodynamics of L-arginine in rats: a model of stimulated neuronal nitric oxide synthesis. AB - Nitric oxide (NO) is believed to be involved in a variety of central nervous system (CNS) functions, including opioid responsivity. Elucidation of the role of NO in the CNS requires the ability to elevate systematically neuronal NO concentrations in vivo. This study was conducted to assess the pharmacokinetics of L-arginine, a NO precursor, and to relate the disposition of this amino acid to the pharmacodynamic endpoint of neuronal NO production. L-Arginine (250-, 500 , or 1000-mg/kg/h) or saline was infused intravenously for 6 h to rats. L Arginine was quantified in brain and blood (after in vivo microdialysis) with high-performance liquid chromatography. NO was quantified simultaneously with a sensitive and specific amperometric sensor placed in the hippocampus. The data were fit with a comprehensive pharmacokinetic-pharmacodynamic (PK/PD) model to obtain parameters governing the systemic disposition of L-arginine, the uptake of L-arginine into the brain, and subsequent NO production. Exogenous administration of L-arginine resulted in incremental elevations in hippocampal NO, with a approximately 33, 48, and approximately 50% increase from control for the 250-, 500-, and 1000-mg/kg/h L-arginine treated rats, respectively. The PK/PD model, which incorporated known characteristics of the system (saturable uptake of L arginine into brain; NO production governed by circadian changes in enzyme activity) was capable of describing accurately the observed data. The model developed herein will be invaluable in characterizing the numerous roles of NO in the CNS. PMID- 14519514 TI - Connectivity of the turtle accessory optic system. AB - Recent whole-cell recordings show that there are multiple synaptic inputs to the accessory optic system of the pond turtle Pseudemys scripta elegans (the basal optic nucleus, BON), suggesting a complex role in visual processing. The BON outputs have now been investigated using transport of diI, rhodamine-conjugated and biotinylated dextrans. Although transport was primarily anterograde, contralateral retinal ganglion cells were labeled retrogradely, confirming that the injection site was a retinal target. Other retrogradely labeled neurons were found ipsilateral to the injection site, in the pretectum, the ventral tegmentum, the dorsal nucleus of the posterior commissure and the lateral habenular nucleus. However, other data indicate that the habenular cells were labeled by spread of the tracer from the BON to the adjacent fasciculus retroflexus and interpeduncular nucleus. Anterogradely labeled fibers projected from BON following three paths, a lateral bundle to the ipsilateral dorsal midbrain, an intermediate bundle to the ipsilateral pretectal area or the posterior commissure and a ventral fiber bundle to the tegmentum bilaterally. Some of these fibers projected caudally through the tegmentum and cerebellar peduncle to terminate just below the Purkinje cell layer of the cerebellar cortex. Fibers that coursed via the intermediate bundle to the posterior commissure were also seen reaching the contralateral pretectal area and the contralateral BON. Injections of the retrograde tracer Fluorogold were also made in the BON to confirm the reciprocal connectivity of both basal optic nuclei. The pathways revealed by these experiments indicate the existence of multiple afferent and efferent connections of the BON, supporting the view that the accessory optic system is more than a simple relay of retinal signals into the brainstem for optokinetic reflexes. PMID- 14519515 TI - Suppression by ethanol of male reproductive activity. AB - Ethanol has been known to suppress reproductive activity in laboratory animals and humans through the inhibition of luteinizing hormone (LH) release by reduction of gonadotropin-releasing hormone (GnRH) secretion from the hypothalamus. There are, however, little data is available regarding the effect of ethanol on GnRH gene expression. Thus, the present study was designed to evaluate the effect of ethanol on GnRH gene expression and reproductive activity at all levels of the hypothalamus-pituitary-gonad (HPG) axis simultaneously. To this end, ethanol (3 g/kg i.p., 15% v/v in saline) was administered to adult male rats for 10 days. Serum levels of LH and testosterone were significantly decreased by ethanol. Using Northern blot analysis and in situ hybridization, the present study showed the reduction in GnRH mRNA levels in the hypothalamus by prolonged ethanol administration. The content of LH in the anterior pituitary was also significantly reduced by ethanol. In addition, steroidogenic acute regulatory protein (StAR) mRNA levels were significantly decreased by ethanol, suggesting a cause for the reduced production of testosterone under this condition. These results indicate that ethanol affects the HPG axis at all the levels. Especially, suppressed GnRH mRNA levels in the hypothalamus of ethanol treated rats strongly demonstrated that hypothalamus is the major action site of ethanol on the HPG axis. Decreased serum LH level may affect the steroidogenesis in the testis, at least in part, through the inhibition of StAR gene expression that induces part of dysfunctions of reproductive activity. PMID- 14519516 TI - Neuroprotective and brain edema-reducing efficacy of the novel cannabinoid receptor agonist BAY 38-7271. AB - BAY 38-7271 is a new high-affinity cannabinoid receptor agonist with strong neuroprotective efficacy in a rat model of traumatic brain injury (acute subdural hematoma, SDH). In the present study we investigated CB1 receptor signal transduction by [35S]GTPgammaS binding in situ and in vitro to assess changes in receptor functionality after SDH. Further, we continued to investigate the neuroprotective properties of BAY 38-7271 in the rat SDH and transient middle cerebral artery occlusion (tMCA-O) model as well as the efficacy with respect to SDH-induced brain edema. [35S]GTPgammaS binding revealed minor attenuation of CB1 receptor functionality on brain membranes from injured hemispheres when compared to non-injured hemispheres or controls. In the rat SDH model, BAY 38-7271 displayed strong neuroprotective efficacy when administered immediately after SDH either as a 1 h (65% infarct volume reduction at 0.1 microg/kg) or short-duration (15 min) infusion (53% at 10 microg/kg). When administered as a 4 h infusion with a 5 h delay after injury, significant neuroprotection was observed (49% at 1.0 microg/kg/h). This was also observed when BAY 38-7271 was administered as a 5 h delayed 15 min short-duration infusion (64% at 3 microg/kg). In addition, the neuroprotective potential of BAY 38-7271 was demonstrated in the rat tMCA-O model, displaying pronounced neuroprotective efficacy in the cerebral cortex (91% at 1 ng/kg/h) and striatum (53% at 10 ng/kg/h). BAY 38-7271 also reduced intracranial pressure (28% at 250 ng/kg/h) and brain water content (20% at 250 ng/kg/h) when determined 24 h post-SDH. Based on these data it is concluded that the neuroprotective efficacy of BAY 38-7271 is mediated by multiple mechanisms triggered by cannabinoid receptors. PMID- 14519517 TI - Modulation of stress-induced dopamine release by excitotoxic damage of the entorhinal cortex in the rat. AB - In the sham-operated rats, exposure to either footshock or psychological stress induced similar biphasic alterations of dopamine (DA) release (an initial increase followed by a decrease below baseline levels) in the amygdala 4 weeks after the surgery. On the other hand, the left entorhinal cortex lesions abolished the late decrement phase of DA release below baseline levels. These results suggest that entorhinal cortex lesions modulate stress-induced dopaminergic transmissions in the lateral amygdala. PMID- 14519518 TI - Interaction between catecholaminergic terminals and urocortinergic neurons in the Edinger-Westphal nucleus in the rat. AB - Central stress regulatory pathways utilize various neurotransmitters/neuropeptides, such as urocortin (Ucn) and catecholamines. Ucn is most abundantly expressed in the Edinger-Westphal nucleus (E-WN), co distributed with catecholaminergic terminals. Acute stress recruits E-WN neurons, and ascending catecholaminergic pathways also contribute to the activation of various brain areas in response to stress. We hypothesized that catecholamine and Ucn interactions in the E-WN mediated the recruitment of these neurons in response to stress. Using double-labeling immunohistochemistry, we found close appositions between urocortin-immunoreactive nervous structures and dopaminergic terminals, however, depletion of them had no effect on the activation pattern of E-WN neurons upon acute immune challenge. From these results we conclude that dopaminergic terminals innervating E-WN Ucn neurons do not play a major role in mediating the responses of E-WN neurons upon acute immune challenge. PMID- 14519519 TI - Glycine mediated alterations in intracellular pH. AB - Glycinergic transmission shapes the coding properties of the lateral superior olivary nucleus (LSO). We investigated intracellular pH responses in the LSO to glycine using BCECF-AM in brain slices. With extracellular bicarbonate, glycine produced an alkalinization followed by an acidification while, in the nominal absence of bicarbonate, glycine produced acidifications. Separately, in whole cell recordings from LSO neurons, glycine caused hyperpolarization followed by long-lasting depolarization. While the bicarbonate-dependent intracellular alkalinization could be related to chloride/bicarbonate exchange, bicarbonate independent acidification may be triggered by depolarization. PMID- 14519520 TI - The monocarboxylate transport inhibitor, alpha-cyano-4-hydroxycinnamate, has no effect on retinal ischemia. AB - Glial-derived monocarboxylate lactate is thought to be an important energy source for neurons during brain activation or in hypoxia-ischemia. Treatment with alpha cyano-4-hydroxycinnamate (4-CIN), a monocarboxylate transporter inhibitor, has been recently reported to exacerbate delayed neuronal damage in a rat model of cerebral ischemia, an effect ascribed to inhibition of lactate/pyruvate transport. Since monocarboxylate transporters are abundant in the retina, we examined the effect of 4-CIN administration on the outcome of high intraocular pressure-induced retinal ischemia in rats. Retinal ischemic damage was assessed by changes in the electroretinogram (ERG), the retinal localization of choline acetyltransferase (ChAT) and neuronal nitric oxide synthase (nNOS) immunoreactivities, and the loss of retinal mRNA for Thy-1. Intraperitoneal or intravitreal administration of 4-CIN had no effect on the ERG or the localization of ChAT and nNOS immunoreactivities in either the control retina or a retina subjected to ischemia/reperfusion. In addition, intravitreal injection of 4-CIN had no effect on ischemia-induced reduction of retinal mRNA levels for Thy-1. These results provide no evidence to support the view that blockade of lactate uptake and/or pyruvate entry into mitochondria for oxidative metabolism has an influence on the outcome of retinal ischemia/reperfusion. PMID- 14519521 TI - Oleic acid induces GAP-43 expression through a protein kinase C-mediated mechanism that is independent of NGF but synergistic with NT-3 and NT-4/5. AB - We have recently shown that the presence of albumin in astrocytes triggers the synthesis and release of oleic acid, which behaves as a neurotrophic factor for neurons. Thus, oleic acid promotes axonal growth, neuronal clustering, and the expression of the axonal growth-associated protein, GAP-43. In this work we show that oleic acid upregulates GAP-43 expression by a protein kinase C (PKC) dependent mechanism. Since GAP-43 expression has been shown to be upregulated by several neurotrophins, we investigated the relationship between the effect of oleic acid and that of NGF, neurotrophin-3 (NT-3) and neurotrophin-4/5 (NT-4/5) on GAP-43 expression. Our results indicate that NGF is not involved in the neurotrophic effect of oleic acid because the addition of NGF did not modify the effect of oleic acid on GAP-43 expression. Neither NT-3 nor NT-4/5 alone modified GAP-43 expression. However, NT-3 and NT-4/5 acted synergistically with oleic acid to increase GAP-43 expression. The lack of effect of NGF as compared to other neurotrophins is not unexpected since we have not found TrkA expression under our experimental conditions. The effect of oleic acid on GAP-43 expression must be independent of autocrine factors synthesized by neurons because this effect was also observed at low cellular densities. In conclusion, our results indicate that oleic acid behaves as a neurotrophic factor, inducing GAP-43 expression through a PKC-mediated mechanism that is not mediated by other neurotrophic factors but that is strongly synergized by NT-3 and NT-4/5. PMID- 14519522 TI - Dietary fatty acids alter blood pressure, behavior and brain membrane composition of hypertensive rats. AB - The beneficial effect of dietary n-3 polyunsaturated fatty acids (PUFAs) on developing hypertension has been repeatedly demonstrated. However, related changes in brain membrane composition and its cognitive correlates have remained unclear. Our study aimed at a comprehensive analysis of behavior and cerebral fatty acid concentration in hypertension after long-term PUFA-rich dietary treatment. Hypertensive and normotensive rats were provided a placebo, or one of two PUFA-enriched diets with a reduced (n-6)/(n-3) ratio for 75 weeks. Exploratory behavior and spatial learning capacity were tested. Systolic blood pressure (BP) was repeatedly measured. Finally, brain fatty acid composition was analyzed by gas chromatography. Hypertensive rats exhibited more active exploration but impaired spatial learning compared to normotensives. Both diets reduced BP, increased PUFA and monounsaturated fatty acid (MUFA) concentration, and reduced saturated fatty acid content in brain. The level of cerebral PUFAs and MUFAs was lower in hypertensive than in normotensive rats. Furthermore, BP positively, while spatial learning negatively correlated with cerebral (n-6)/(n 3) PUFA ratio. We concluded that regular n-3 PUFA consumption could prevent the development of hypertension, but reached only a very delicate improvement in spatial learning. Furthermore, we consider a potential role of metabolically generated MUFAs in the beneficial effects of PUFA supplementation. PMID- 14519523 TI - Group III secreted phospholipase A2 causes apoptosis in rat primary cortical neuronal cultures. AB - Secreted phospholipase A(2) (sPLA(2)) has been shown to cause both necrotic and apoptotic neuronal injury in the central nervous system. The acute effects of group III sPLA(2) (sPLA(2)-III) include an ability to potentiate glutamate induced necrosis and calcium influx in primary rat neuronal cultures. Thus, a brief exposure to glutamate and sPLA(2)-III results in an immediate potentiated calcium influx and a potentiated necrosis the following day after treatment, compared to addition of either agonist alone. In contrast to these acute effects, sPLA(2) from both groups IB and IIA have been shown to induce neuronal cell death via apoptosis. Here, we offer the first evidence that group III sPLA(2) potently causes extensive neuronal apoptosis in rat cortical cultures, as evidenced by DNA fragmentation. Distinct from this effect, even high concentrations of glutamate did not cause neuronal damage under the same conditions, and there was no evidence of sPLA(2) damage to cortical astrocytes. PMID- 14519524 TI - Distribution and morphological characterization of phosphate-activated glutaminase-immunoreactive neurons in cat visual cortex. AB - Phosphate-activated glutaminase (PAG) is the major enzyme involved in the synthesis of the excitatory neurotransmitter glutamate in cortical neurons of the mammalian cerebral cortex. In this study, the distribution and morphology of glutamatergic neurons in cat visual cortex was monitored through immunocytochemistry for PAG. We first determined the specificity of the anti-rat brain PAG polyclonal antibody for cat brain PAG. We then examined the laminar expression profile and the phenotype of PAG-immunopositive neurons in area 17 and 18 of cat visual cortex. Neuronal cell bodies with moderate to intense PAG immunoreactivity were distributed throughout cortical layers II-VI and near the border with the white matter of both visual areas. The largest and most intensely labeled cells were mainly restricted to cortical layers III and V. Careful examination of the typology of PAG-immunoreactive cells based on the size and shape of the cell body together with the dendritic pattern indicated that the vast majority of these cells were pyramidal neurons. However, PAG immunoreactivity was also observed in a paucity of non-pyramidal neurons in cortical layers IV and VI of both visual areas. To further characterize the PAG immunopositive neuronal population we performed double-stainings between PAG and three calcium-binding proteins, parvalbumin, calbindin and calretinin, to determine whether GABAergic non-pyramidal cells can express PAG, and neurofilament protein, a marker for a subset of pyramidal neurons in mammalian neocortex. We here present PAG as a neurochemical marker to map excitatory cortical neurons that use the amino acid glutamate as their neurotransmitter in cat visual cortex. PMID- 14519525 TI - Herpes virus-mediated preproenkephalin gene transfer in the ventral striatum mimics behavioral changes produced by olfactory bulbectomy in rats. AB - The syndrome of behavioral, physiological, and neurochemical changes caused by ablation of the olfactory bulbs (OBX) in rats serves as a reliable and well validated model of depression. Previous experiments have demonstrated that OBX leads to increased expression of the preproenkephalin (ENK) gene in the olfactory tubercle (OT) portion of the ventral striatum in rats. The aim of the present experiments was to investigate the role of OBX-induced ENK overexpression in the OT in the behavioral abnormalities exhibited by bulbectomized rats. A recombinant herpes virus carrying human preproENK cDNA was used to manipulate ENK gene expression in the OT of bulbectomized and sham-operated rats. Motivational deficits were assessed by the sucrose preference test, and 'agitation-like' behaviors were measured with the novel open field and footshock-induced freezing tests. ENK gene transfer in sham-operated rats mimicked some of the effects of OBX; it decreased freezing behavior in response to mild footshock and produced behavioral activation in the open field. In another experiment, virally mediated ENK gene transfer into the OT of intact rats decreased footshock-induced freezing, and this effect was reversed by naltrexone administration. PreproENK gene transfer into the OT did not produce analgesic effects in the tail-flick test. No effects on freezing behavior were observed following preproENK gene transfer into the frontal cortex. An additional experiment revealed that naltrexone administration attenuated the OBX-induced abnormality in freezing behavior. The results indicate that overexpression of the preproENK gene in the ventral striatum may mediate the 'agitation-like' behavior exhibited by bulbectomized rats. PMID- 14519526 TI - Novel effects on memory observed following unilateral intracranial administration of okadaic acid, cyclosporin A, FK506 and [MeVal4]CyA. AB - The involvement of protein phosphatases and peptidyl-prolyl cis/trans isomerases (PPIases) in memory formation in the chick has previously been investigated using a single-trial learning task. In these studies, inhibitory agents were administered bilaterally directly to a critical area of the chick brain. These studies are now extended to investigate whether similar effects are obtained if the drugs are administered unilaterally. All of the effects reported previously following bilateral administration of okadaic acid (OA), cyclosporin A (CyA), FK506 and [MeVal(4)]CyA can be attributed to their action in just one hemisphere. OA, at a concentration known to selectively inhibit PP2A in vitro (0.5 nM) results in permanent memory loss from 30-40 min post-training when injected in the left hemisphere, but has no effect when injected in the right hemisphere. A higher concentration of OA (100 nM), which inhibits both PP2A and PP1 in vitro, has a similar effect in the left hemisphere but causes a transient period of memory loss from 10-20 min post-training when injected in the right hemisphere. CyA (5 nM and 20 nM), which inhibits both PP2B and PPIase activity, causes permanent memory loss from 60 min post-training when injected into the left hemisphere, an effect also observed following administration of FK506 (20 nM), which also inhibits PP2B and PPIase activity, and [MeVal(4)]CyA (5 nM), which inhibits PPIase activity but not PP2B activity. Administration of CyA (20 nM) and FK506, but not [MeVal(4)]CyA, in the right hemisphere leads to a transient period of memory loss from 10-20 min post-training. These results confirm significant roles for phosphatases and PPIases in memory processing but challenge previous conclusions drawn on the basis of bilateral drug administration protocols. PMID- 14519527 TI - Serum amyloid P component induces neuronal apoptosis and beta-amyloid immunoreactivity. AB - Previously we have reported serum amyloid P component (SAP) induced cell death in cerebro-cortical cultures of rat brain. In this paper we studied the types of target cells and the molecular mechanism of SAP-induced cell death. Immuno electron and light microscopy revealed that SAP penetrates the plasma membrane and translocates selectively into the nuclei of neurons. Neuronal cells with SAP immunoreactivity exhibit the morphological hallmarks of apoptosis in vitro. The apoptotic mechanism of cell death is also supported by the increased Bax/Bcl-2 ratio. In addition to neurotoxic effects, we detected elevated beta-amyloid (Abeta) immunoreactivity following SAP treatment. This study supports the thesis that SAP plays an important role in the pathomechanism of neurodegenerative diseases, including Alzheimer's disease by inducing neuronal apoptosis. PMID- 14519528 TI - Inhibitors of nitric oxide synthase block carbon monoxide-induced increases in cGMP in retina. AB - Previous studies indicate that the gaseous messengers carbon monoxide (CO) and nitric oxide (NO) can interact to cause robust increases in intracellular cGMP levels in the retina. The purpose of the present study was to investigate the biochemical basis of the interactions between NO and CO for these increases. Turtle retinas were incubated in vitro with CO to stimulate cGMP production in the presence or absence of the nitric oxide synthase inhibitors N-omega-nitro-L arginine methyl ester and S-methyl-thiocitrulline. Cyclic GMP immunocytochemistry was then used to evaluate the changes in cGMP levels in response to these stimuli. The results indicated that CO itself stimulated increases in cGMP in bipolar and amacrine cells, and that the increases were completely blocked by SMTC and L-NAME. We postulate that the increases of cGMP in response to CO might be mediated, at least partly, by CO displacing and releasing NO from its intracellular storage pool(s). PMID- 14519529 TI - Estrogen receptors and metabolic activity in the human tuberomamillary nucleus: changes in relation to sex, aging and Alzheimer's disease. AB - The human tuberomamillary nucleus (TMN), that is the sole source of histamine in the brain, is involved in arousal, learning and memory and is impaired in Alzheimer's disease (AD) as shown by the presence of cytoskeletal alterations, a reduction in the number of large neurons, a diminished neuronal metabolic activity and decreased histamine levels in the hypothalamus and cortex. Experimental data and the presence of sex hormone receptors suggest an important role of sex steroids in the regulation of the function of TMN neurons. Therefore, we investigated sex-, age- and Alzheimer-related changes in estrogen receptor alpha and beta (ERalpha and ERbeta) in the TMN. In addition, metabolic activity changes of TMN neurons were determined by measuring Golgi apparatus (GA) and cell size. In the present study, ERalpha immunocytochemical expression in AD patients did not differ from that in elderly controls. However, a larger amount of cytoplasmic ERbeta was found in the TMN cells of AD patients. Earlier studies, using the GA size as a parameter, have shown a clearly decreased metabolic activity in the TMN neurons in AD. In the present study, the size of the GA did not change during aging, indicating the absence of strong metabolic changes. Cell size of the TMN neurons appeared to increase during normal aging in men but not in women. Concluding, the enhanced cytoplasmic expression of ERbeta in the TMN may be involved in the diminished neuronal metabolism of these neurons in AD patients. PMID- 14519530 TI - Intrathecal injection of cAMP response element binding protein (CREB) antisense oligonucleotide attenuates tactile allodynia caused by partial sciatic nerve ligation. AB - The transcription factor cAMP responsive element binding protein (CREB) is important in regulating immediate-early genes and some late-effector genes involved in neuroplasticity in response to peripheral injury and stressful insults. Partial nerve injury elicited neuropathic pain is accompanied by increased phosphorylation of CREB in the ipsilateral spinal cord dorsal horn (Ma and Quirion, Pain 93 (2001) 295; Miletic et al., Pain 99 (2002) 493). The aim of this study is to determine whether increased phosphorylation of CREB in the dorsal horn contributes to the pathogenesis of neuropathic pain. Three weeks following partial sciatic nerve ligation (PSNL), daily intrathecal injection of antisense CREB oligodeoxynucleotide (ODN, 20 microg/day) for 5 days significantly attenuated tactile allodynia. The attenuation lasted for more than 4 days. Total CREB and phosphorylated CREB in both ipsilateral and contralateral dorsal horn neurons were dramatically reduced in antisense ODN injected PSNL rats 1 week after injection. The extent of reduction of total CREB and phosphorylated CREB containing cells in the dorsal horn ipsilateral to injury was greater than in the contralateral dorsal horn. These data suggest that phosphorylation of CREB is an important contributing event in the central plasticity of nerve injury and in the pathogenesis of neuropathic pain. PMID- 14519531 TI - Functional characterisation of the active ascorbic acid transport into cerebrospinal fluid using primary cultured choroid plexus cells. AB - Crossing the blood-CSF barrier is an important pathway for certain nutrients to enter the CNS. Cultured choroid plexus epithelial cells are a potent model system to study active transport properties of this tissue in vitro. In the present study this in vitro model was used to analyse ascorbic acid transport across the blood-CSF barrier that is supposedly mediated by the Na(+)-dependent transporter SVCT2. The expression of SVCT2 in the cultured cells was proven by RT-PCR. Active transport across the cell monolayer resulted in ascorbic acid enrichment at the CSF mimicking side. Ascorbic acid transport and uptake were decreased to 13 and 27%, respectively, in the presence of 200 microM phloretin. Inhibition of both transepithelial substrate transport (to 7.5%) and cytoplasmatic uptake (to 20%) was observed in Na(+)-free medium indicating that a basolaterally located and Na(+)-dependent transporter mediates ascorbic acid uptake. Substituting Cl(-) by either iodide or D-gluconate increased ascorbic acid uptake by factors of 3.7 or 2.5, respectively. Similar observations were made when Na(+)-dependent myo inositol transport was analysed. Additionally, in presence of 100 microM bumetanide, an inhibitor of Na(+)-Cl(-)-cotransport, indirectly increased ascorbic acid and myo-inositol transport rates were observed showing that ascorbic acid-Na(+)-cotransport might balance low intracellular Na(+) concentration. PMID- 14519532 TI - Polysynaptic slow depolarization and spiking activity elicited after induction of long-term potentiation in rat auditory cortex. AB - Polysynaptic activity was recorded in supragranular pyramidal neurons before and after the induction of long-term potentiation (LTP) in slices obtained from rat auditory cortex. LTP was induced by tetanic stimulation of layer IV. In the pyramidal neurons exhibiting LTP, repetitive stimulation at 50 Hz with 15 pulses triggered a slow 15-35 mV depolarization lasting 0.5-2 s with two to five spike discharges. There was no such response before the induction of LTP or in the neurons that did not exhibit LTP. Slow depolarization with spike discharges was blocked by an NMDA receptor antagonist but not by a metabotropic glutamate receptor antagonist. The reversal potential of the slow depolarization was approximately -7 mV and the membrane resistance decreased during slow depolarization, suggesting that the slow depolarization was produced by polysynaptic excitatory post-synaptic potentials. LTP was also induced by low frequency stimulation paired with a depolarizing current injection. In the pyramidal neurons exhibiting LTP after the paired stimulation, the slow depolarization amplitude was small and repetitive stimulation did not trigger spike discharges. Tetanic stimulation is expected to induce LTP in the polysynaptic neural circuits connecting many pyramidal neurons. The present findings suggest that polysynaptic activity can be generated in the potentiated neural circuits. Such activity might serve to read out the memory stored in polysynaptic neural circuits in the cerebral cortex. PMID- 14519533 TI - Presence of beta-arrestin-1 immunoreactivity in the cutaneous nerve fibers of rat glabrous skin. AB - beta-Arrestin-1 (betaArr1) plays a major role in the desensitization and internalization of G protein-coupled receptors. We previously localized betaArr1 in the sensory neurons of rat lumbar 4 and 5 dorsal root ganglia (DRG) and reported the predominant presence of betaArr1 in the small-diameter DRG neurons that are often implicated with nociception. Because of betaArr1's crucial role in regulating the initiation of cellular signaling, in the current study we evaluated the distribution of betaArr1 in the peripheral sensory terminals where various receptors are present. Western blotting confirmed the presence of betaArr1 immunoreactivity in the rat skin. Sciatic nerve ligation demonstrated that betaArr1 is transported peripherally from the DRG, and immunohistochemistry showed betaArr1 immunoreactivity in the glabrous skin of the rat hindpaw. In the glabrous skin, strong betaArr1 immunoreactivity was detected in nerve fibers in the dermal nerve plexus and dermal papillae. Fine varicose immunoreactive fibers were found in the epidermis. In addition, betaArr1 was observed in specialized sensory receptors such as Meissner corpuscles. Our observations thus indicate that betaArr1 may be involved in modulation of specific tactile stimulation from the skin in addition to nociception. PMID- 14519534 TI - The effects of ageing and of DSP-4 administration on the micturition characteristics of male Wistar rats. AB - This study sought to determine the effects of ageing on the in vivo micturition characteristics of male Wistar rats and to assess whether they might be replicated in young rats by using the neurotoxin DSP-4 to lesion locus coeruleus derived noradrenergic pathways projecting to spinal cord nuclei controlling micturition. Significant age-related changes in micturition patterns were observed. There was a loss of a diurnal rhythm in micturition patterns and a large increase in voided volume, maximal between 21 and 24 months, which was paralleled by an increased water intake. DSP-4 lesions neither altered micturition patterns nor water intake in the young adult rat. DSP-4 induced changes in the pattern of tyrosine hydroxylase-like immunoreactivity (TH-LI), most notably almost complete depletion of TH-LI in the dorsolateral nucleus and retention of TH-LI in lumbosacral autonomic preganglionic nuclei, did not mimic the changes in the pattern of TH-LI seen in aged rats. PMID- 14519535 TI - Medial preoptic area dopaminergic responses to female pheromones develop during puberty in the male Syrian hamster. AB - Chemosensory cues from receptive females do not elicit similar reactions before and after puberty in male hamsters. While pheromones facilitate a complex display of reproductive behavior in adults, prepubertal males do not engage in these same behaviors. Dopamine (DA) released from the medial preoptic area (MPOA) in response to a receptive female or her odors is an important component of the neural events underlying adult male rat sexual behavior. The current experiment investigated whether increased dopaminergic activity occurs in the adult male hamster MPOA in response to female pheromones, and if so, whether this response is absent in prepubertal males, which do not mate. Sexually nai;ve prepubertal and adult male hamsters were exposed to cotton swabs with or without pheromone from an estrous female for 0, 5, 15, or 25 min, after which brains were collected and frozen on dry ice. The MPOA was micropunched from frozen coronal sections (500 microm), and concentrations of DA and its primary metabolite DOPAC were determined by high-performance liquid chromatography-electrochemical detection. DOPAC was used as an index of dopaminergic activity. DOPAC levels significantly increased in adults after 15 min exposure to pheromone. In contrast, MPOA DOPAC concentrations did not increase in prepubertal males exposed to pheromone. These data demonstrate that the neural processing of sexually relevant chemosensory stimuli matures during puberty. The absence of a DA response to female pheromones prior to puberty may contribute to the inability of prepubertal males to display reproductive behavior. PMID- 14519536 TI - The roles of different types of glutamate receptors involved in the mediation of nucleus submedius (Sm) glutamate-evoked antinociception in the rat. AB - Based on our previous findings that glutamate microinjected into the thalamic nucleus submedius (Sm) inhibits dose-dependently the rat tail-flick (TF) reflex, this study investigated which glutamate receptor subtype is involved in mediating this effect. The effects of an NMDA (N-methyl-D-aspartate), non-NMDA or metabotropic glutamate receptor (mGluR) antagonist microinjected into Sm on the TF reflex were examined in untreated or in Sm glutamate treated (microinjection into the Sm) rats. The TF latencies were measured in each of these groups of rats every 5 min. Injection of DNQX [6,7-dinitroquinoxaline-2,3(1H,4H)-dione], a non NMDA receptor antagonist, or (+/-)-MCPG [(+/-)-alpha-methyl-4 carboxyphenylglycine], a mGluR antagonist, into the Sm blocked the inhibitory effects induced by a subsequent microinjection of glutamate into the same Sm site. The TF latency increased only by 6.6+/-1.6 or 9.0+/-1.1%, respectively, of the baseline value, which was markedly less than that (51.3+/-8.4 or 50.7+/-5.3%) obtained from injection of glutamate only (P<0.001, n=8). However, pre microinjection of MK-801 [(+)-5-methyl-10,11-dibenzo[a,d]cyclohepten-5,10-imine], an NMDA receptor antagonist, into the Sm had no effect on the Sm glutamate-evoked inhibition of the TF reflex. The TF latency change (40.0+/-11.1%) was not significantly different (P>0.05, n=8) compared with that obtained from glutamate injection alone. These observations suggest that non-NMDA and metabotropic glutamate receptors, but not NMDA receptors, are involved in mediating Sm glutamate-evoked antinociception. PMID- 14519537 TI - ACh and ATP mediate excitatory transmission in cat carotid identified chemoreceptor units in vitro. AB - Several molecules have been proposed as excitatory transmitters between glomus (type 1) cells and nerve terminals of petrosal ganglion (PG) neurons in the carotid body (CB). We tested whether ACh and ATP have a role to play as excitatory transmitters in the cat CB by recording intracellularly from identified PG neurons functionally connected to the CB in vitro. PG neurons projecting to the CB were classified according to their intracellular responses as: (a) neurons with humped action potentials (hAP neurons) responding phasically to long-lasting depolarizing pulses (53/67), and (b) neurons with smooth action potentials (non-hAP neurons) that fire tonically during long-lasting depolarizations (14/67). CB stimulation by stop flow and/or acidosis induced activity in 28 of 39 hAP-type neurons, being classified as chemosensory, but in none of the non-hAP neurons. Hexamethonium (10 microM) and suramin (100 microM) reversibly abolished the increased discharges evoked in chemosensory neurons (8/9) by stop flow or acidosis. Moreover, 24 of 27 chemosensory neurons responded to ganglionar application of ACh and ATP, while two neurons responded only to ACh and one to ATP. Mechanical deformation of the carotid sinus induced firing activity in 10 of 13 non-hAP neurons, but in none of the hAP neurons tested. Interestingly, 4/10 non-hAP neurons, which responded to carotid sinus mechanical stimulation also responded to ganglionar application of ATP, but were insensitive to ACh. Present results favor the hypothesis that ACh and ATP are excitatory transmitters in the cat CB, acting-at least-on the PG neuron terminals in the CB. PMID- 14519538 TI - Impact of adolescent nicotine exposure on adenylyl cyclase-mediated cell signaling: enzyme induction, neurotransmitter-specific effects, regional selectivities, and the role of withdrawal. AB - Recent animal studies indicate that the adolescent brain is especially vulnerable to nicotine-induced alterations in synaptic function, echoing the increased susceptibility to nicotine dependence and withdrawal noted for adolescent smokers. We administered nicotine to adolescent rats via continuous minipump infusions from PN30 to PN47.5, using 6 mg/kg/day, a dose rate that replicates the plasma nicotine levels found in smokers, and examined the effects on cell signaling mediated through adenylyl cyclase (AC) and its response to catecholamines. Studies were conducted during nicotine administration (PN45) and in the posttreatment, withdrawal period (PN50, 60, 75). Adolescent nicotine augmented AC activity as evidenced by increased responsiveness to the direct AC stimulants, forskolin and Mn(2+). The effects on AC were equally noted in brain regions enriched (striatum) or sparse (cerebellum) in cholinergic projections, implying that the effects are secondary to activation/repression of neural circuits, rather than representing direct effects on AC mediated by nicotinic cholinergic receptors. AC responses to dopaminergic and noradrenergic stimulants were also enhanced by nicotine exposure. However, in contrast to earlier work with serotonin-mediated responses, the effects on catecholaminergic stimulation were smaller and did not display the sex-dependence noted for serotonin. An alternate administration paradigm that maximizes episodic withdrawal (twice-daily nicotine injections) induced AC more rapidly at lower nicotine doses. Our results indicate that adolescent nicotine exposure elicits lasting alterations in synaptic signaling that intensify and persist during withdrawal. These findings support the concept that the adolescent brain is especially susceptible to persistent nicotine-induced alterations. PMID- 14519539 TI - Antioxidant activity of X-34 in synaptosomal and neuronal systems. AB - Inhibiting aggregation and deposition of amyloid beta-peptide (Abeta) in brain is a therapeutic strategy for Alzheimer's disease (AD). A Congo-red-like molecule, X 34, is reported to bind to Abeta deposits. Oxidative stress associated with Abeta is hypothesized to be critical for the neurotoxic properties of this peptide. The present study was undertaken to test the hypothesis that X-34, with its salicylate groups, would act as an antioxidant. When challenged by hydroxyl or peroxyl free radicals or Abeta(1-42), oxidative stress and neurotoxicity occurred in neural systems as assessed by several indices. However, pretreatment of synaptosomes and primary neuronal cell culture with X-34 greatly ameliorated lipid peroxidation induced by these free radicals and Abeta(1-42). Protein oxidation was not prevented by X-34. These results are discussed in terms of potential therapeutic use of X-34 and related compounds in AD. PMID- 14519540 TI - Immunocytochemical analysis of glutamate and GABA in hippocampus of genetic absence epilepsy rats (GAERS). AB - In the present study, we used an immunocytochemical technique at the electron microscopic level to determine if there are changes in the glutamate and GABA neurotransmitter content of the hippocampus of genetic absence epilepsy rats from Strasbourg (GAERS). We also investigated if there was mossy fiber reorganization. After perfusion fixation, brains were removed and cryostat sections were stained according to the neo-Timm's procedure. High-resolution electron microscopy was used for ultrastructural examination of the hippocampus of GAERS and non epileptic control Wistar animals. For ultrastructural and immunocytochemical studies, ultrathin-cut sections were obtained and immunolabeled with anti glutamate and anti-GABA antibodies. The number of gold particles per nerve terminal was counted and the area of the nerve terminal was determined using the program NIH Image Analysis. No mossy fiber sprouting was detected in the hippocampus of GAERS. GABA and glutamate immunoreactivity were observed in the mossy fiber terminals of both the control and GAERS groups. Glutamate density in the CA3 region of GAERS hippocampus was found to be significantly increased compared to the control group. However, there was no difference in the GABA density of nerve terminals and in areas of GABAergic and mossy terminals between GAERS and the control group. The difference in glutamate level may merely be due to strain differences between the GAERS strain and the original Wistar strain or it is also possible that it appears after seizures have started. PMID- 14519541 TI - Effect of intrathecal nocistatin on nociceptin/orphanin FQ analgesia in chronic constriction injury rat. AB - Nocistatin and nociceptin/orphanin FQ (N/OFQ) are two neuropeptides derived from the same precursor protein, prepronociceptin (ppOFQ), and exhibit different effects on spinal neurotransmission. Nocistatin does not bind to nociceptin/orphanin FQ peptide receptor (NOP), but intrathecal (i.t.) nocistatin has been reported to block the analgesic effect of i.t. N/OFQ. In this study, we investigated the effect of i.t. nocistatin on N/OFQ analgesia to radiant thermal stimuli in chronic constriction injury (CCI) rat. Firstly, to investigate the analgesic effect of N/OFQ, different doses of N/OFQ (3, 10, 30 microg) were intrathecally injected and foot withdrawal latency (FWL) to radiant heat was recorded. It is observed that 3 microg N/OFQ had no effect on FWL, 10 and 30 microg N/OFQ significantly increased FWL of CCI rat. Then, 10 microg N/OFQ, 10 microg nocistatin and a drug cocktail including 10 microg N/OFQ and 10 microg nocistatin were intrathecally injected. The results showed that FWL significantly decreased after using N/OFQ and nocistatin compared with using only N/OFQ, and 10 microg nocistatin had no effect on FWL versus control, suggesting that this dose of nocistatin per se had no effect on the pain threshold of CCI rat, but could block the analgesic effect of N/OFQ. These results indicated that i.t. N/OFQ dose relatedly depressed thermal hyperalgesia produced by CCI and nocistatin could block N/OFQ analgesia at spinal level in CCI rat. PMID- 14519542 TI - NADPH oxidase immunoreactivity in the mouse brain. AB - Superoxide production via NADPH oxidase has been shown to play a role in neurotoxicity, ischemic stroke, and possibly Parkinson's and Alzheimer's diseases. In addition, NADPH oxidase-dependent production of superoxide may be necessary for normal brain functions, including neuronal differentiation and neuronal plasticity. To improve our understanding of NADPH oxidase in the brain, we studied the localization of the various protein components of NADPH oxidase in the central nervous system of the adult mouse using immunohistochemistry. We detected staining for the cytoplasmic NADPH proteins, p40(phox), p47(phox), and p67(phox), as well as the membrane-associated NADPH oxidase proteins, p22(phox) and gp91(phox) in neurons throughout the mouse brain. Staining of each of the NADPH oxidase proteins was observed in neurons in all regions of the neuraxis, with particularly prominent localizations in the hippocampus, cortex, amygdala, striatum, and thalamus. The expression of NADPH oxidase proteins in neurons suggests the possibility that enzymatic production of superoxide by a NADPH oxidase may play a role in both normal neuronal function as well as neurodegeneration in the brain. PMID- 14519543 TI - The impact of motor activity on intracerebral ERPs: P3 latency variability in modified auditory odd-ball paradigms involving a motor task. AB - The P3 wave of event-related potentials was recorded with intracranial electrodes in 24 epileptic patients during the pre-surgical evaluation of epilepsy surgery. Three different cognitive auditory paradigms were used: (1) odd-ball paradigm with no output required (PGI) where patients had simply to recognize target tones, (2) odd-ball with motor response (PGII), where patients had to press a button in response to target tones, and (3) odd-ball with both counting task and motor response (PGIII), where patients had to recognize target tones, press a button in response to them, and count their number. The occurrence of P3 potential, its latency and amplitude, and the dependence of P3 latency on the task complexity were calculated. Identifiable P3 potentials in all the three paradigms were recorded from locations in mesial cortex (18 locations mesial temporal, eight locations mesial frontal, two locations mesial parietal) and lateral sites (eight sites lateral temporal, five lateral frontal, and two lateral parietal). P3 latency values ranged from 257 to 320 ms in all explored cortical areas when PGI was used; they significantly increased or decreased during PGII and PGIII, depending on the task and structure explored. In the mesial temporal cortex, the changes of P3 latency between paradigms were minimal. In the mesial parietal cortex, there was significant P3 delay in both PGII and PGIII relative to PGI. In the mesial frontal cortex, there was a significant latency decrease in PGII, and practically identical mean latency in PGI and PGIII. In all lateral cortices (temporal, frontal and parietal), there was always a P3 latency increase in PGII and PGIII relative to PGI, the most significant results being observed in the parietal and frontal lateral areas. The results support the multi-generator theory of P3. Prolongation of the mean P3 latency in lateral frontal and parietal cortices when the paradigm involved the execution of a motor task might reflect specific gating in this area during active movements, while the absence of modification in the temporal lobe may reflect minimal involvement of this region in motor planning or processing. The prolongation of mean P3 latency in practically all lateral structures in PGIII suggests that most cortical areas were involved in the cognitive functions needed for this test. The finding of reduction and subsequent prolongation of P3 latency in the mesial frontal cortex might reflect the unique specialization of this area and its specific involvement in motor processing. PMID- 14519544 TI - Effect of deep brain stimulation of GPI on neuronal activity of the thalamic nucleus ventralis oralis in a dystonic patient. AB - OBJECTIVE: To record the possible effect of acute deep brain stimulation (DBS) of the globus pallidus internus (GPI) on the neuronal activity of the ventralis oralis anterior (VOA) nucleus of the thalamus. METHODS: Under general propofol anaesthesia, extracelullar single unit recordings were performed in VOA of a post anoxic dystonic patient previously implanted with GPI located electrodes for chronic DBS. RESULTS: Neurons recorded in the VOA could be classified in two cell subpopulations: a high firing rate (16.5 Hz) and low burst index (BI; 15.6) type and a low firing rate (5.5 Hz) and high BI (35.6) type. GPI electrical stimulation reduced the frequency and increased the BI of the high firing rate cells while leaving the other cell type unchanged. CONCLUSION: These results demonstrate that pallidal DBS is able to inhibit a subpopulation of motor thalamic cells and question the pathophysiological model of dystonia based on a low firing rate of GPI cells. PMID- 14519545 TI - Magnetoencephalographic and electroencephalographic evaluation in patients with cryptogenetic partial epilepsy. AB - Magnetoencephalography (MEG) has been applied for more than 20 years to the localization of the epileptic focus in partial epilepsies, but correlation with electroencephalographic (EEG) data in homogeneous groups of patients is scarce. OBJECTIVE: The aim of our work was to use EEG and MEG for the study of a group of adults and children affected by cryptogenetic partial epilepsy. METHODS: We analyzed the traces obtained from electroencephalographic and magnetoencephalographic recordings of 10 patients of ages ranging from 7 to 38 years affected by cryptogenetic partial epilepsy. We evaluated the presence of commonly detected or uniquely detected spikes, and, whenever possible, we used MEG for localization of the epileptic focus. RESULTS: Three patients showed no epileptic activity during the EEG and MEG sessions. Overall agreement between EEG and MEG (presence of concordant spikes with the same localization shown by both techniques) was obtained in five patients. In one patient the spikes detected by EEG and MEG were different, and in another patient interictal activity was demonstrated exclusively by EEG. CONCLUSIONS: EEG in this series was not inferior to MEG in terms of spike detection. Combination of EEG and MEG is feasible, better than each technique alone, and may be useful for non-invasive diagnosis and monitoring of pediatric and adult patients with partial epilepsies. PMID- 14519546 TI - Acute low single dose of apomorphine reduces periodic limb movements but has no significant effect on sleep arousals: a preliminary report. AB - STUDY OBJECTIVES: To investigate the effects of apomorphine on the frequency of periodic limb movements during sleep (PLMS) and on sleep architecture. DESIGN: Nine patients presenting PLMS (including eight patients with restless legs syndrome) underwent three consecutive night sleep recordings. They received a single dose of 0.5 mg subcutaneous apomorphine at bedtime the third night. RESULTS: When computing PLMS during four 2-h periods of sleep, a significant period by apomorphine-effect was demonstrated, with a marked reduction of PLMS during the first 4 h post-injection (P < 0.01). No significant differences were found in sleep macroarchitecture between the three recorded nights, excepted a slight reduction in sleep latency during the third night (P < 0.05). Despite the decreased number of PLMS after apomorphine injection, there were significant changes neither in the total number of arousals nor in the index of arousals per hour of sleep. CONCLUSION: Our results add further support to the dopaminergic hypothesis in the generation of PLMS. The persistence of arousals suggests that they are not simply the consequence of PLMS but a primary phenomenon, not related with the dopaminergic system. PMID- 14519547 TI - [Symmetry and temporal variability of neurography]. AB - The aims of the present study are to document side-to-side differences and temporal variability, between two trials (T1 and T2 at a time interval of 3 months) of nerve conduction measurements collected from 30 healthy subjects (mean age 22 +/- 2 years). METHODS: The protocol at T1 consisted of motor nerve conduction studies of median, ulnar, peroneal and tibial nerves bilaterally, with measurement of (a) motor response size (amplitude and area); (b) terminal latency; (c) minimal, mean and maximal F-wave latency; (d) motor conduction velocity; and (e) F-wave occurrence. T1 also involved sensory nerve conduction studies of median, ulnar, radial, lateral and medial cutaneous, sural and superficial peroneal nerves bilaterally, with measurement of sensory potential size (amplitude and area) and computation of sensory conduction velocity. The protocol at T2 consisted of identical measurements from the dominant side. RESULTS AND CONCLUSION: There was a negative relationship between the variability of parameters evaluating nervous conduction and the length of the nerve segment under study. Thus, the smallest side-to-side and temporal variabilities are measured for minimal F-wave latencies (on average 2-3%). The limits of symmetry and temporal variability are particularly useful for diagnosis of unilateral peripheral neuropathy or neurophysiological follow-up of patients with neuropathy, when the variability of the parameter under study is weak and when there is a high correlation between values recorded on the left and on the right or at T1 and T2. This was the case for motor response size of tibial and ulnar nerves, sensory potential size of radial nerve and minimal F-wave latencies from each studied motor nerve. PMID- 14519548 TI - Peripheral chemoreceptor activity in sleeping neonates exposed to warm environments. AB - In neonates, it is often assumed that ventilatory control and heat stress interact. Thus the two factors have been implicated in various pathologies (apnoea, sudden infant death syndrome). However, little is known about the mechanisms of this interaction, and the influence of sleep is still debated. This study aimed at determining the influence of warm exposure on the decrease in ventilation during a hyperoxic test (HT), which is considered to be a measure of peripheral chemoreceptor activity. The test was performed in active (AS) and quiet sleep (QS) in 12 neonates exposed to thermoneutral or warm environments. The HT consisted of 30 s of inspired, 100% O(2). The ventilatory response was assessed in terms of a response time, defined as the time elapsing between HT onset and the first significant change in V(E). Our results show that, in both thermal conditions, the fall in V(E) was higher in AS than in QS. Warm exposure significantly enhanced the ventilatory response in AS (-27.5 +/- 8.7% vs. -38.3 +/- 8.8%, P < 0.01) but not in QS. A thermometabolic drive or inputs from thermoreceptors could be involved in the reinforcement of peripheral chemoreceptor activity in AS in warmer environments, which could contribute to an increasing risk of apnoea in neonates with altered chemoreceptor function. Since hypothalamic structures are involved in thermoregulatory, sleep processes and (probably) in respiratory control, it could well be the principal site where this interaction occurs. PMID- 14519549 TI - Mesial temporal lobe epilepsy: a pathological replay of developmental mechanisms? AB - Focal epilepsies of the mesial temporal lobe are often associated with a hippocampal sclerosis. Changes in Cl homeostasis and GABAergic signalling in downstream regions, may contribute to the human pathology. A review of changes in cellular and synaptic function and connectivity after de-afferentation suggests this epileptic syndrome may involve a pathological replay of developmental mechanisms. PMID- 14519550 TI - How the confocal laser scanning microscope entered biological research. AB - A history of the early development of the confocal laser scanning microscope in the MRC Laboratory of Molecular Biology in Cambridge is presented. The rapid uptake of this technology is explained by the wide use of fluorescence in the 80s. The key innovations were the scanning of the light beam over the specimen rather than vice-versa and a high magnification at the level of the detector, allowing the use of a macroscopic iris. These were followed by an achromatic all reflective relay system, a non-confocal transmission detector and novel software for control and basic image processing. This design was commercialized successfully and has been produced and developed over 17 years, surviving challenges from alternative technologies, including solid-state scanning systems. Lessons are pointed out from the unusual nature of the original funding and research environment. Attention is drawn to the slow adoption of the instrument in diagnostic medicine, despite promising applications. PMID- 14519551 TI - The propagation speeds of calcium action potentials are remarkably invariant. AB - This paper critically compiles all published cases of established or putative calcium action potentials (or ultrafast calcium waves) where their speeds are known and are not limited by intercellular delays. The 127 cases include data from neurons or nerve nets within systems that range from cnidaria, ctenophores, molluscs, crustaceans, worms, echinoderms and tunicates up to mammalian brains; from muscle cells within organisms that range from Beroe, Cestum, moths, a crab, molluscs, a tunicate, frogs, chick embryos and turtles up to mammalian hearts; from epithelia in cnidaria and tunicates; even from a dinoflagellate and an insectivorous plant as well as reconstituted heart strands. They reveal a restriction to values of about 10-40 cm/sec at 20 degrees C and comparable restrictions at other temperatures. Moreover--unlike the speeds of sodium action potentials--the speeds of calcium ones are unrelated to cell diameter, at least over the available range of about 0.1 to 30 microns. Why do calcium action potentials have such fixed propagation speeds? Perhaps evolution has driven them to be the fastest waves of calcium influx which avoid subsurface poisoning. PMID- 14519552 TI - Long term in vitro-cultured plant cells show typical neoplastic features at the cytological level. AB - Cells from a green normal (dependent on exogenous hormones) callus and from an achlorophyllous fully habituated (independent from exogenous hormones) callus, both generated from the same sugarbeet strain more than twenty years ago, were reexamined cytologically, ten years after the first comparative description. Cells from the habituated callus, already considered as neoplastic cells, because terminating a neoplastic progression where the organogenic totipotency was lost, still showed nuclear invaginations, polynucleolation, vacuolation of nucleoli and incomplete cell walls, nevertheless at a higher degree. The present study particularly shows that, compared to their previous description, normal cells have started to acquire some features (polynucleolation, nuclear invaginations.) that are typical of the neoplastic cells. This suggests that normal cells, on the long term, also entered a neoplastic progression, which should explain the known progressive loss of regeneration capacity of too long subcultured hormone dependent calli. PMID- 14519553 TI - Duplexes of 21-nucleotide RNAs mediate RNA interference in differentiated mouse ES cells. AB - RNA interference (RNAi) is a novel phenomenon that can induce post transcriptional gene silencing (PTGS) both in animals and plants. RNAi is effective in suppressing specific gene expression in the early mouse embryonic cells and in undifferentiated embryonic stem (ES) cells. In this study, we demonstrate that dsRNA is effective in inducing PTGS in differentiated ES cells: CD34+ embryoid body (EB) cells, as confirmed by western blot and immunocytochemical staining. PU.1 is a key transcription factor in myeloid differentiation. Undifferentiated ES cells do not express PU.1; however it is expressed when ES cells differentiate into EBs. PU.1 could be suppressed by the specific PU.1 dsRNA, but not non-specific Lamin A/C dsRNA, in the CD34+ EB cells when they were induced to myeloid differentiation in the presence of GM-CSF and IL-3. As a consequence, the level of expression of CD115 (M-CSF receptor), one of the downstream genes regulated by PU.1 is decreased in PU.1 dsRNA treated CD34+ EB cells, but not in Lamin A/C dsRNA treated cells. To explore this phenomenon in other myeloid gene, we also found that C/EBPalpha gene could be knocked down by C/EBPalpha dsRNA. Our finding demonstrates that RNAi is effective in inhibiting specific gene expression in differentiated ES cells. RNAi offers a new methodology for study of hematopoietic regulation using ES cell differentiation. PMID- 14519554 TI - An in vitro study of the sub-cellular distribution of nicotinic receptors. AB - Nicotinic and serotoninergic 5HT3 receptors share important sequence identities except for their cytoplasmic loop. Both ends of this loop display conserved 3D helical structures with distinct primary sequences. We decided to check whether these two helices named F and G play a role in the sub-cellular distribution of different nicotinic receptors. We systematically exchanged each helix with the equivalent sequence of neuronal nicotinic and alpha4, beta2 and alpha7 subunits in the functional chimeric alpha7-5HT3 receptor used as a model system. The new chimeras were expressed in vitro in polarized epithelial cells from pig kidney. We quantified synthesis and export of the receptors to the cell surface by measuring alpha-bungarotoxin binding sites. Immunogold labelling was used, at the electron microscope level, to determine the amount of each chimera present at either domain, apical and/or basolateral, of these cells. We noticed that in epithelial cells the majority of alpha-bungarotoxin binding sites remained sequestered in the cytoplasm as already observed in neurons in vivo. The majority of the pentamers present at the cell surface were located at the apical domain. Our results suggest that helix F and G differently regulate assembly and export to the cell surface of alpha-bungarotoxin binding receptors. PMID- 14519555 TI - Ultrastructure of auxiliary and gonimoblast cells during carposporophyte development in the red alga Cryptopleura ruprechtiana (Delesseriaceae, Ceramiales, Rhodophyta). AB - The ultrastructure sequence for the complete post-fertilization development is described in Cryptopleura ruprechtiana (C. Agardh) Kylin, a member of the Delesseriaceae. Following fertilization the diploid nucleus is transferred to the auxiliary cell. This contains typical red algal proplastids, cytoplasmic concentric membranes, numerous small vacuoles and lipid bodies. Crystalline inclusions and virus-like particles are also present. In addition darkly staining spherical masses possibly represent dehydrated haploid chromatin. The multinucleate auxiliary cell produces initially one large gonimoblast initial and subsequently many smaller gonimoblast initials. The first formed generative gonimoblast cell is similar in cellular structure to the auxiliary cell. Gonimoblast initials are uninucleate but through caryokinesis they become multinucleate. They undergo repeated cleavage to form more gonimoblast cells. Subsequent, centripetal cytokinesis results in the formation of clusters of gonimoblast cells. A new type structural cap or association is observed in the septal plugs that interconnect gonimoblast initials. Terminal or generative gonimoblast cells cleave to form additional gonimoblast cells. Only terminal gonimoblast cells are differentiated to carpospores. PMID- 14519556 TI - Use of cryo-negative staining in tomographic reconstruction of biological objects: application to T4 bacteriophage. AB - Recent advances in electron microscopy and image analysis techniques have resulted in the development of tomography, which makes possible the study of structures neither accessible to X-ray crystallography nor nuclear magnetic resonance. However, the use of tomography to study biological structures, ranging from 100 to 500 nm, requires developments in sample preparation and image analysis. Indeed, cryo-electron tomography present two major drawbacks: the low contrast of recorded images and the sample radiation damage. In the present work we have tested, on T4 bacteriophage samples, the use of a new preparation technique, cryo-negative staining, which reduces the radiation damage while preserving a high signal-to-noise ratio. Our results demonstrate that the combination of cryo-negative staining in tomography with standard cryo-microscopy and single particle analysis results in a methodological approach that could be useful in the study of biological structures ranging in the T4 bacteriophage size. PMID- 14519557 TI - Myosin heavy chain isoforms in postnatal muscle development of mice. AB - In this study, using a high-resolution gel electrophoresis technique, we have characterized the myosin heavy chain composition in different skeletal muscle of the mouse during postnatal development. The pattern of myosin heavy chain expression was studied in four hind limb muscles, the diaphragm, the tongue and the masseter. All of these muscles displayed the usual sequential transitions from embryonic to neonatal and to adult myosin heavy chain isoforms but more interestingly these transitions occur with a distinct chronology in the different muscles. In addition, our results demonstrated a transitory pattern of expression for certain adult myosin heavy chain isoforms in the soleus and the tongue. In the soleus muscle IIB and in the tongue IIA myosin heavy chain isoforms were detected only for a short time during postnatal life. Our results demonstrate that muscles of the mouse with different functions are subjected to a distinct programs of myosin isoform transitions during postnatal muscle development. This study describes new data which will help us to understand both postnatal muscle development in transgenic mouse muscles as well as in muscle pathology. PMID- 14519558 TI - Effect of taxol and okadaic acid on microtubule dynamics in thimerosal-arrested primary mouse oocytes: a confocal study. AB - A pulse of thimerosal (TMS), a sulfhydryl reagent, induces an instantaneous, complete and long-lasting microtubule interphasic network disassembly in mouse primary oocytes, correlated with the irreversible inhibition of meiosis reinitiation This inhibition is bypassed by dithiothreitol (DTT) while thiosalicylic acid, an analog of TMS, does induce neither microtubules depolymerisation nor inhibition of reinitiation and resumption of meiosis. This strongly suggests that the dramatic and pleiotropic inhibitory effect of TMS is specifically related to its sulfhydryl group oxidising activity of critical molecules among which tubulin. In contrast to DTT, okadaic acid (OA), known to bypass the inhibitory effect of drugs interfering with protein kinase activities, induces a late chromatin condensation and GVBD in TMS-pulsed oocytes as compared to the control situation, with no significant concomitant microtubule assembly. These cytological features are suggested to be indirectly induced by a late MAPK activation and confirm that a very early thiol oxidation induced by TMS exerts a much more dramatic effect on resumption of meiosis than any pharmacological manipulation of protein kinase activities leading to activation of MPF. Finally, taxol was shown to promote tubulin polymerisation even when microtubules were irreversibly disassembled by thiol oxidation but fails to restore the ability to undergo maturation. PMID- 14519559 TI - Comparative insights and shared directions for adolescent health: a commentary on three international papers. PMID- 14519560 TI - Co-occurrence of health-risk behaviors among Beijing middle school students, China. PMID- 14519561 TI - Effects of peer education on reproductive health knowledge for adolescents living in rural areas of Turkey. PMID- 14519562 TI - Prevention and health promotion in school and community settings: a commentary on the international perspective. PMID- 14519563 TI - Adolescent health, school health activities, community contexts, and health surveys in Sweden. AB - School health activities have been very important in improving adolescent health in Sweden for almost 200 years. In the 1800s, emphasis was on medical services. Vaccination programs and medical examinations became the key issues. Deterioration of adolescent health in the 1960s changed the objectives of both school education and health services to health promotion. Important members of the community followed suit and involvement of the local community has remained a hallmark, even though the extent and direction varies. The subsequent period was characterized by substantial improvement in adolescent health behavior. The latter years of the 1980s and the 1990s saw deterioration of adolescent health behavior. Less emphasis on health promotion, decentralization of school health responsibility from physicians to administrators, and heavy savings directed toward schools were important mediators. Adolescents were also more engaged in international youth cultures with liberal practices, such as drug use. Community surveys of adolescent health behaviors have proven to be important in mobilizing broad local involvement in adolescent health promotion. The lesson learned is that health promotion has to involve as many community members as possible. Coordinating resources and having unified objectives is cost efficient PMID- 14519564 TI - Changing schools, changing health? Design and implementation of the Gatehouse Project. AB - PURPOSE: To describe the Gatehouse Project which addresses the social context of high school with an aim of changing students' sense of school connection and in turn, health risk behavior and well-being. METHODS: Distinguishing features of the project were its conceptual framework, implementation process, and evaluation design. The conceptual framework derived from attachment theory and focused on three aspects of the school social context: security, communication, and participation. Implementation was standardized around a survey of the school social environment, creation of a school-based action team, and the implementation of strategies matched to a school's profile of need. In addition, an early high school curriculum addressed the skills relevant to social functioning and emotional adjustment. The evaluation design was based on a cluster randomized trial involving 26 schools. It used follow-up of an individual cohort and repeat cross-sectional surveys to capture outcomes at an individual student and whole-school level. RESULTS AND CONCLUSIONS: The Gatehouse Project drew on both health and education research to develop and coordinate a broad based school health promotion intervention. It represents a promising new direction for school-based preventive work. PMID- 14519565 TI - International developments in adolescent health care: a story of advocacy and achievement. AB - The contemporary health problems of young people occur within the context of the physical, social, cultural, economic, and political realities within which they live. There are commonalities and differences in this context among developed and developing countries, thus differing effects on the individual's personal as well as national development. Internationally, the origins and evolution of health care for adolescents can be viewed as an unfolding saga taking place particularly over the past 30 years. It is a story of advocacy and subsequent achievement in all corners of the world. This paper reviews the important developments in the international arena, recognizes major pioneers and milestones, and explores some of the current and future issues facing the field. The authors draw heavily on their experiences with the major nongovernmental adolescent health organizations. The special roles of the World Health Organization, Pan American Health Organization, and United Nations Children's Fund (UNICEF) are highlighted, and special consideration is given to the challenge of inclusion through youth participation. PMID- 14519566 TI - Health-related quality of life in youth: a comparison between adolescents and young adults with uncomplicated epilepsy and healthy controls. AB - PURPOSE: To describe health-related quality of life (HRQOL) in adolescents and young adults with uncomplicated epilepsy and to compare it with a random sample of the general population. METHODS: Young people, aged 13-22 years, meeting the criteria and registered in four Swedish hospitals answered questionnaires (n = 158/193) on HRQOL together with 282 (n = 282/390) random controls living in the same areas in Sweden. The instruments used were "I think I am," "Youth Self Report," "Sense of coherence," and "Family APGAR." Data were analyzed by using factorial analysis of variance. RESULTS: Girls had a poorer HRQOL than boys. The epilepsy group reported lower competence (i.e., they were less active, had lower social competence and poorer school achievement). Both groups had an overall positive self-esteem. Differences between girls in the epilepsy and control groups were small, whereas differences among boys were more evident. Older age was related to poorer HRQOL in both groups. CONCLUSIONS: This study points out the importance of being observant of signs of stigmatization in adolescents with epilepsy. PMID- 14519567 TI - Does making clinic-based reproductive health services more youth-friendly increase service use by adolescents? Evidence from Lusaka, Zambia. AB - PURPOSE: To report the findings of a study that evaluated the impact of three youth-friendly service (YFS) projects in Lusaka, Zambia. In 1994, the Lusaka District Health Management Team (LDHMT) identified adolescents as a priority underserved population with regard to reproductive health information and services. As part of its long-term goal to improve the health and well-being of Lusaka youth, the LDHMT, in collaboration with CARE, UNICEF/Zambia Family Life Movement, and John Snow International, implemented three separate YFS projects to increase service use among adolescents. METHODS: Service statistics from 10 clinics (8 "treatment clinics" and 2 "non-YFS clinics") were used to measure adolescent service use. Qualitative and quantitative data were collected to measure the degree of "youth-friendliness" at the clinics and the level of community acceptance of providing reproductive health services to youth. Specific indicators of youth-friendliness were developed that measured the attitudes of the clinic staff toward giving services to youth, whether clinic staff honored privacy and confidentiality, whether boys and young men were welcomed, whether the clinic policies supported providing services for youth, whether clinic staff promoted its services to youth in surrounding community, and whether youth, themselves, perceived that they would be welcomed and have their needs met at the clinics. Similarly, indicators of community acceptance were developed that measured whether parents and other adults supported the provision of reproductive health services to youth. RESULTS AND CONCLUSIONS: Although the projects appear to have improved the clinic experience for adolescent clients and to have increased service use levels at some clinics, the findings suggest that community acceptance of reproductive health services for youth may have a larger impact on the health-seeking behaviors of adolescents. PMID- 14519568 TI - Health and health care-seeking behavior of adolescents in urban and rural China. AB - A cross-sectional survey was conducted among 13-18-year-old adolescents in Zhejiang Province, China. Morbidities showed few urban/rural differences. Concerns were mainly related to academic pressure. Respondents were eager users of health services; 73% were taking regular medication, but 52% had never attended a dentist. PMID- 14519569 TI - Socioeconomic patterning of smoking, sedentary lifestyle, and overweight status among adolescents in Norway and the United States. AB - Increased efforts are needed to promote adolescent health in all areas investigated in this study. Differences both in prevalence and socioeconomic patterning between Norway and the United States indicate that the major challenges differ between the two countries. The results also emphasize the need to undertake joint efforts to use measurements that make national data sets comparable across countries. PMID- 14519570 TI - Adolescent substance use: beware of international comparisons. AB - PURPOSE: To compare prevalence estimates of adolescents' cigarette, alcohol and marijuana use from one Australian and two U.S. surveys, and to consider the effect of methodological differences on reported use. METHODS: Secondary analysis of data from the Australian Child and Adolescent Component of the National Survey of Mental Health and Well-being (NSMHWB, 1998), the U.S. Youth Risk Behavior Survey (YRBS, 1999), and National Household Survey on Drug Abuse (NHSDA, 1998). Prevalence estimates and 95% confidence intervals were derived for three substance use behaviors. Differences between estimates were considered statistically significant if the 95% confidence intervals did not overlap. RESULTS: When Australian and U.S. adolescents were compared using NSMHWB and YRBS data, the former were generally found to be less likely to report using cigarettes, alcohol and marijuana than the latter. However, when NHSDA was used as the comparator, the prevalence of substance use among Australian adolescents was either no different from, or greater than, that of U.S. adolescents. Likely explanations for the discrepant findings include the population focus (i.e., whether the survey targeted only young people or adults as well), sampling issues (i.e., whether school-based and household-based sampling frames introduced different biases in terms of who was likely to be absent when the survey was administered), response rates, the survey context (i.e., whether school-based and household-based survey administration methods promoted different kinds of response behavior), the wording of questions and the precision of estimates of the different surveys. CONCLUSIONS: Cross-national data on adolescent substance use should be interpreted cautiously. Cross-national comparisons that are done well (i.e., using standard, uniform approaches) can be invaluable in highlighting worthwhile policy directions; cross-national comparisons that are done poorly may lead to erroneous assumptions. PMID- 14519571 TI - Implementation and evaluation of an internet health site for adolescents in Switzerland. AB - Ciao is a website specifically designed for young people and focuses mainly on health issues. This report presents the process of setting up the site and a first evaluation undertaken by using two self-administered questionnaires administered via the website itself. It suggests that it is possible to provide young people with authoritative health information and to facilitate their access to counseling and health care facilities by having young people use such a website. PMID- 14519572 TI - Thyroid hemiagenesis. PMID- 14519573 TI - Transition to adult health care for adolescents and young adults with chronic conditions: position paper of the Society for Adolescent Medicine. PMID- 14519574 TI - Species concepts: what for? PMID- 14519575 TI - Tick "talk": protein release by tick salivary cells. PMID- 14519576 TI - How many pathways for invasion of the red blood cell by the malaria parasite? AB - Merozoites of the malaria parasite Plasmodium falciparum use several receptors for cellular engagement when they invade human red blood cells. Recently, a merozoite erythrocyte-binding protein, EBA-140, has been identified that specifically binds to glycophorin C on red blood cells. Up to 50% of Melanesians have a deletion in this gene, and the resultant Gerbich-negative red blood cells are relatively resistant to invasion. While discovery of multiple pathways for invasion could confound the search for suitable vaccine targets, they could also be considered in the design of therapeutic interventions that prevent malaria parasites entering red blood cells. PMID- 14519577 TI - Chemoresistance in falciparum malaria. PMID- 14519579 TI - Comparative genomics to uncover the secrets of tsetse and livestock-infective trypanosomes. PMID- 14519580 TI - The pig strain of Echinococcus granulosus in humans: a neglected issue? PMID- 14519581 TI - Encystation-specific vesicles in Giardia: a primordial Golgi or just another secretory compartment? PMID- 14519582 TI - Chagas disease: a role for autoimmunity? PMID- 14519583 TI - Mass administrations of antimalarial drugs. PMID- 14519584 TI - CD36 and malaria: friends or foes? PMID- 14519585 TI - DNA microarrays in parasitology: strengths and limitations. AB - Genome sequencing efforts have provided a wealth of new biological information that promises to have a major impact on our understanding of parasites. Microarrays provide one of the major high-throughput platforms by which this information can be exploited in the laboratory. Many excellent reviews and technique articles have recently been published on applying microarrays to organisms for which fully annotated genomes are at hand. However, many parasitologists work on organisms whose genomes have been only partially sequenced and where little, if any, annotation is available. The focus of this review is on how to use and apply microarrays to these situations. PMID- 14519586 TI - Essential elements of postabortion care: origins, evolution and future directions. PMID- 14519588 TI - Moving from research to program--the Egyptian postabortion care initiative. PMID- 14519587 TI - Comparing the quality of three models of postabortion care in public hospitals in Mexico City. AB - CONTEXT: Each year, an estimated 120,000 women in Mexico seek treatment in public hospitals for abortion-related complications--the country's fourth leading cause of maternal mortality. Models of postabortion care emphasizing counseling and provision of contraceptives have the potential to improve the quality of care these women receive. METHODS: Between April 1997 and August 1998, women treated for abortion complications in six Mexican Institute of Social Security (IMSS) hospitals in the Mexico City metropolitan area were surveyed. Data related to patient-provider interaction, information provision and counseling were analyzed for three models of care: sharp curettage standard care, sharp curettage postabortion care and manual vacuum aspiration postabortion care. RESULTS: Women in the two postabortion care groups rated the quality of services they received more highly than did those receiving sharp curettage standard care. A significantly greater proportion of women treated under the postabortion care models than of those treated under the sharp curettage standard model received information about their health status before treatment, the uterine evacuation procedure, signs of postabortion complications and care at home. In addition, a greater proportion of women treated under the postabortion care models accepted a contraceptive method before leaving the facility (64-78% vs. 40%). CONCLUSIONS: Implementation of a postabortion care model contributes to the delivery of high quality services to women experiencing abortion complications. The standard IMSS model of postabortion treatment should be modified to emulate those in hospitals that systematically link general counseling and family planning services to the clinical services provided to women with abortion complications. PMID- 14519589 TI - Client-provider communication in postabortion care. PMID- 14519590 TI - Facility-level reproductive health interventions and contraceptive use in Uganda. AB - CONTEXT: In Uganda, modern contraceptive use has recently increased in areas served by the Delivery of Improved Services for Health (DISH) project. Whether these increases are associated with facility-level factors is unknown, however. METHODS: Data from the 1999 DISH Evaluation Surveys were used in multivariate logistic regressions to assess the independent relationships of five indicators of the family planning service environment with individual-level use of a modern contraceptive in rural and urban areas. The surveys consisted of a household questionnaire of 1,766 women of reproductive age and a facility module implemented in all health facilities that serve the sampled population. RESULTS: After women's social and demographic characteristics were controlled for, none of the service environment factors was independently associated with current use of a modern method in rural areas. By contrast, in urban areas, the proximity of a private health facility (which likely reflects an increased availability of methods) was positively associated with current use (odds ratio, 2.1), as was the presence of a higher number (three or more) of DISH-trained service providers (1.7). CONCLUSIONS: The presence of private health facilities was the factor most strongly associated with contraceptive use in urban areas, perhaps because they improved the availability of methods. Few other facility-level program inputs had significant effects. PMID- 14519591 TI - The impact of household delivery of family planning services on women's status in Bangladesh. AB - CONTEXT: Qualitative studies assessing the impact of the household delivery of family planning services on women's social status have yielded contradictory findings. Given the resumption of these services in Bangladesh in 2003, it is important to reevaluate the social impact of the doorstep program using quantitative techniques. METHODS: Longitudinal and cross-sectional data from 3,783 women using doorstep services in two rural districts of Bangladesh are used in ordinary least-squares and logistic regression analyses to assess the effect of doorstep services on changes in women's status between 1988 and 1993. RESULTS: In analyses controlling for background characteristics, women's status in 1988, previous service use and visit selection bias, household outreach is associated with increases in women's status between 1988 and 1993. However, this effect is largely attributable to the impact of doorstep services on women's ability to regulate their fertility rather than to the home visits themselves. CONCLUSION: The decision of the Bangladesh Ministry of Health and Family Welfare to resume household family planning services should not be detrimental to women's status, and may be associated with gender benefits to female clients served by the program. PMID- 14519592 TI - Local anesthetics worsen renal function after ischemia-reperfusion injury in rats. AB - Local anesthetics are widely used during the perioperative period, even in patients with preexisting renal disease. However, local anesthetics have been shown to cause cell death in multiple cell lines, including human kidney proximal tubule cells. We questioned whether local anesthetics potentiate renal dysfunction after ischemia-reperfusion (I/R) injury in vivo. Rats were implanted with subcutaneous miniosmotic pumps that continuously delivered lidocaine (2 mg.kg-1.h-1), bupivacaine (0.4 mg.kg-1.h-1), tetracaine (1 mg.kg-1.h-1), or saline vehicle, and 6 h later the rats were subjected to 30 min of renal ischemia or to sham operation. Renal function was assessed by measurement of plasma creatinine at 24 and 48 h after renal I/R injury in the presence or absence of chronic infusions of local anesthetics and correlated to histological changes indicative of necrosis. The degree of renal apoptosis was assessed by three methods: 1) DNA fragmentation detected by terminal deoxynucleotidyl transferase biotin-dUTP nick-end labeling staining, 2) DNA laddering detected after agarose gel electrophoresis, and 3) morphological identification of apoptotic tubules at the corticomedullary junction. We also measured the expression of the proinflammatory markers ICAM-1 and TNF-alpha. Continuous local anesthetic infusion with renal I/R injury resulted in an increased magnitude and duration of renal dysfunction compared with the saline-infused I/R group. Additionally, both apoptotic and necrotic renal cell death as well as inflammatory changes were significantly potentiated in local anesthetic-treated rat kidneys. Local anesthetic infusion alone without I/R injury had no effect on renal function. We conclude that local anesthetics potentiated renal injury after I/R by increasing necrosis, apoptosis, and inflammation. PMID- 14519593 TI - Inhibition of endocytosis causes phosphorylation (S256)-independent plasma membrane accumulation of AQP2. AB - Inhibition of clathrin-mediated endocytosis by expression of a GTPase-deficient dynamin mutant (dynamin-2/K44A) for 16 h results in an accumulation of plasma membrane aquaporin-2 (AQP2) in epithelial cells stably transfected with wild-type AQP2. We now show a similar effect of K44A dynamin in LLC-PK1 cells transfected with an S256 phosphorylation-deficient AQP2 mutant, AQP2(S256A), and in AQP2 transfected inner medullary collecting duct (IMCD) cells. More acute blockade of endocytosis in these cells with the cholesterol-depleting agent methyl-beta cyclodextrin (mbetaCD; 10 mM) resulted in a rapid and extensive cell-surface accumulation of both wild-type AQP2 and AQP2 (S256A) within 15 min after treatment. This effect was similar to that induced by treatment of the cells with vasopressin. Blockade of endocytosis by mbetaCD was confirmed using quantitative analysis of FITC-dextran uptake and AQP2 membrane insertion was verified by cell surface biotinylation. These data indicate that AQP2 recycles constitutively and rapidly between intracellular stores and the cell surface in LLC-PK1 and IMCD cells. The constitutive trafficking process is not dependent on phosphorylation of the serine-256 residue of AQP2, which is, however, an essential step for regulated vasopressin/cAMP-mediated translocation of AQP2. Our data show that rapid and extensive plasma membrane accumulation of AQP2 can occur in a vasopressin receptor (V2R)- and phosphorylation-independent manner, pointing to a potential means of bypassing the mutated V2R in X-linked nephrogenic diabetes insipidus to achieve cell surface expression of AQP2. PMID- 14519594 TI - Suppressions of chronic glomerular injuries and TGF-beta 1 production by HGF in attenuation of murine diabetic nephropathy. AB - Diabetic nephropathy is now the leading cause of end-stage renal diseases, and glomerular sclerotic injury is an initial event that provokes renal dysfunction during processes of diabetes-linked kidney disease. Growing evidence shows that transforming growth factor-beta 1 (TGF-beta 1) plays a key role in this process, especially in eliciting hypertrophy and matrix overaccumulation. Thus it is important to find a ligand system to antagonize the TGF-beta 1-mediated pathogenesis under high-glucose conditions. Herein, we provide evidence that hepatocyte growth factor (HGF) targets mesangial cells, suppresses TGF-beta 1 production, and minimizes glomerular sclerotic changes, using streptozotocin induced diabetic mice. In our murine model, glomerular sclerogenesis (such as tuft area expansion and collagen deposition) progressed between 6 and 10 wk after the induction of hyperglycemia, during a natural course of diabetic disease. Glomerular HGF expression levels in the diabetic kidney transiently increased but then declined below a basal level, with manifestation of glomerular sclerogenesis. When anti-HGF IgG was injected into mice for 2 wk (i.e., from weeks 4 to 6 after onset of hyperglycemia), these glomerular changes were significantly aggravated. When recombinant HGF was injected into the mice for 4 wk (i.e., between 6 and 10 wk following streptozotocin treatment), the progression of glomerular hypertrophy and sclerosis was almost completely inhibited, even though glucose levels remained unchanged (>500 mg/dl). Even more important, HGF repressed TGF-beta 1 production in glomerular mesangial cells even under hyperglycemic conditions both in vitro and in vivo. Consequently, not only albuminuria but also tubulointerstitial fibrogenesis were attenuated by HGF. Overall, HGF therapy inhibited the onset of renal dysfunction in the diabetic mice. On the basis of these findings, we wish to emphasize that HGF plays physiological and therapeutic roles in blocking renal fibrogenesis during a course of diabetic nephropathy. PMID- 14519595 TI - Three-dimensional functional reconstruction of inner medullary thin limbs of Henle's loop. AB - Digital three-dimensional (3-D) functional reconstructions of inner medullary nephrons were performed. Antibodies against aquaporins (AQP)-1 and -2 and the chloride channel ClC-K1 identified descending thin limbs (DTLs), collecting ducts (CDs), and ascending thin limbs (ATLs), respectively, through indirect immunofluorescence. Tubules were labeled in transverse sections and assembled into 3-D arrays, permitting individual tubule or combined surface representations to depths of 3.3 mm to be viewed in an interactive digital model. Surface representations of 75 tubules positioned near the central region of the inner medulla were reconstructed. In most DTL segments that form loops below 1 mm from the inner medullary base, AQP1 expression begins at the base, becomes intermittent for variable lengths, and continues nearly midway to the loop. The terminal DTL segment exhibiting undetectable AQP1 represents nearly 60% of the distance from the medullary base to the tip of the loop. AQP1 expression was entirely undetectable in shorter long-looped DTLs. ClC-K1 is expressed continuously along the terminal portion of all DTLs reconstructed here, beginning with a prebend region approximately 164 microm before the bend in all tubules and continuing through the entire ascent of the ATLs to the base of the inner medulla. CDs express AQP2 continuously and extensive branching patterns are illustrated. 3-D functional reconstruction of inner medullary nephrons is capable of showing axial distribution of membrane proteins in tubules of the inner medulla and can contribute to further development and refinement of models that attempt to elucidate the concentrating mechanism. PMID- 14519596 TI - Pravastatin treatment attenuates interstitial inflammation and fibrosis in a rat model of chronic cyclosporine-induced nephropathy. AB - We investigated the effects of pravastatin, a competitive inhibitor of 3-hydroxy 3-methylglutaryl-coenzyme A (HMG-CoA) reductase, on interstitial inflammation and fibrosis, using an animal model of chronic cyclosporine A (CsA)-induced nephropathy. Sprague-Dawley rats were maintained on a low-salt diet (0.05% sodium) and treated daily for 1 or 4 wk with vehicle (olive oil; 1 ml/kg sc), CsA (15 mg/kg sc), or both CsA and pravastatin (5 or 20 mg/kg in the drinking water). Anti-inflammatory and antifibrotic effects of pravastatin were studied by evaluating the concentrations of the inflammatory mediators osteopontin (OPN) and C-reactive protein (CRP), of fibrotic cytokine-transforming growth factor (TGF) beta1, and the presence of ED-1-positive cells (macrophages). In addition, renal function, serum lipid levels, histopathology (arteriolopathy and tubulointerstitial fibrosis), and the expression of the vasoactive factors endothelial nitric oxide synthase (eNOS) and renin protein were also compared for different treatment groups. Pravastatin induced dose-dependent decreases in the expression of OPN, intrarenal CRP, and TGF-beta1, and in the numbers of ED-1 positive cells at 1 and 4 wk. These were accompanied by a significant attenuation of tubulointerstitial fibrosis at 4 wk. The downregulation of eNOS protein expression in CsA-treated rat kidney was markedly upregulated by pravastatin treatment, although intrarenal renin expression was unaffected. Renal dysfunction induced by CsA significantly improved with administration of pravastatin at a dose of 20 mg/kg. Neither CsA nor pravastatin influenced serum lipid or high sensitivity CRP levels in the treatment groups. Thus in chronic CsA nephropathy, pravastatin effectively abrogates the progression of tubulointerstitial inflammation and fibrosis. This may support the clinical use of pravastatin. PMID- 14519597 TI - Human skeletal muscle PPARalpha expression correlates with fat metabolism gene expression but not BMI or insulin sensitivity. AB - Peroxisome proliferator-activated receptor-alpha (PPARalpha) is a key regulator of fatty acid oxidation in skeletal muscle, but few data exist from humans in vivo. To investigate whether insulin sensitivity in skeletal muscle and body mass index (BMI) were associated with skeletal muscle expression of PPARalpha and with important genes regulating lipid metabolism in humans in vivo, we undertook hyperinsulinemic-euglycemic clamps and measured PPARalpha mRNA levels and mRNA levels of lipid regulating PPARalpha response genes in skeletal muscle biopsies. mRNA levels were measured in 16 men, using a novel highly sensitive and specific medium throughput quantitative competitive PCR that allows reproducible measurement of multiple candidate mRNAs simultaneously. mRNA levels of PPARalpha were positively correlated with mRNA levels of CD36 (r = 0.77, P = 0.001), lipoprotein lipase (r = 0.54, P = 0.024), muscle-type carnitine palmitoyltransferase-I (r = 0.54, P = 0.024), uncoupling protein-2 (r = 0.63, P = 0.008), and uncoupling protein-3 (r = 0.53, P = 0.026), but not with measures of insulin sensitivity, BMI, or GLUT4, which plays an important role in insulin mediated glucose uptake. Thus our data suggest that in humans skeletal muscle PPARalpha expression and genes regulating lipid metabolism are tightly linked, but there was no association between both insulin sensitivity and BMI with PPARalpha expression in skeletal muscle. PMID- 14519598 TI - Unlike mice, dogs exhibit effective glucoregulation during low-dose portal and peripheral glucose infusion. AB - Portal infusion of glucose in the mouse at a rate equivalent to basal endogenous glucose production causes hypoglycemia, whereas peripheral infusion at the same rate causes significant hyperglycemia. We used tracer and arteriovenous difference techniques in conscious 42-h-fasted dogs to determine their response to the same treatments. The studies consisted of three periods: equilibration (100 min), basal (40 min), and experimental (180 min), during which glucose was infused at 13.7 micromol.kg(-1).min(-1) into a peripheral vein (p.e., n = 5) or the hepatic portal (p.o., n = 5) vein. Arterial blood glucose increased approximately 0.8 mmol/l in both groups. Arterial and hepatic sinusoidal insulin concentrations were not significantly different between groups. p.e. exhibited an increase in nonhepatic glucose uptake (non-HGU; Delta8.6 +/- 1.2 micromol.kg( 1).min(-1)) within 30 min, whereas p.o. showed a slight suppression (Delta-3.7 +/ 3.1 micromol.kg(-1).min(-1)). p.o. shifted from net hepatic glucose output (NHGO) to uptake (NHGU; 2.5 +/- 2.8 micromol.kg-1.min-1) within 30 min, but p.e. still exhibited NHGO (6.0 +/- 1.9 micromol.kg(-1).min(-1)) at that time and did not initiate NHGU until after 90 min. Glucose rates of appearance and disappearance did not differ between groups. The response to the two infusion routes was markedly different. Peripheral infusion caused a rapid enhancement of non-HGU, whereas portal delivery quickly activated NHGU. As a result, both groups maintained near-euglycemia. The dog glucoregulates more rigorously than the mouse in response to both portal and peripheral glucose delivery. PMID- 14519599 TI - Reduced beta-cell mass and altered glucose sensing impair insulin-secretory function in betaIRKO mice. AB - Pancreatic beta-cell-restricted knockout of the insulin receptor results in hyperglycemia due to impaired insulin secretion, suggesting that this cell is an important target of insulin action. The present studies were undertaken in beta cell insulin receptor knockout (betaIRKO) mice to define the mechanisms underlying the defect in insulin secretion. On the basis of responses to intraperitoneal glucose, approximately 7-mo-old betaIRKO mice were either diabetic (25%) or normally glucose tolerant (75%). Total insulin content was profoundly reduced in pancreata of mutant mice compared with controls. Both groups also exhibited reduced beta-cell mass and islet number. However, insulin mRNA and protein were similar in islets of diabetic and normoglycemic betaIRKO mice compared with controls. Insulin secretion in response to insulin secretagogues from the isolated perfused pancreas was markedly reduced in the diabetic betaIRKOs and to a lesser degree in the nondiabetic betaIRKO group. Pancreatic islets of nondiabetic betaIRKO animals also exhibited defects in glyceraldehyde- and KCl-stimulated insulin release that were milder than in the diabetic animals. Gene expression analysis of islets revealed a modest reduction of GLUT2 and glucokinase gene expression in both the nondiabetic and diabetic mutants. Taken together, these data indicate that loss of functional receptors for insulin in beta-cells leads primarily to profound defects in postnatal beta cell growth. In addition, altered glucose sensing may also contribute to defective insulin secretion in mutant animals that develop diabetes. PMID- 14519600 TI - Acknowledgement of psychiatric research funding. PMID- 14519601 TI - Post-concussion syndrome: clarity amid the controversy? PMID- 14519602 TI - Suicide: the leading cause of maternal death. PMID- 14519603 TI - Understanding the suicidal brain. PMID- 14519604 TI - Jaspers was right after all--delusions are distinct from normal beliefs. For. PMID- 14519605 TI - Defining thought broadcast. Semi-structured literature review. AB - BACKGROUND: The symptom of thought broadcast may have diagnostic significance but its definition varies. AIMS: To examine multiple definitions of thought broadcast in different texts, to synthesise their common features and to undertake local and national surveys of psychiatrists to determine which definitions they endorse. METHOD: A semi-structured literature review of electronic databases, supplemented by a manual search of psychiatric textbooks, conceptual analyses and postal surveys of clinicians in North Trent (58 trainees and 70 consultants) and throughout the UK (49 professors of general adult psychiatry). RESULTS: Thought broadcast is susceptible to multiple definitions: three exemplars were identified in the literature, each endorsed by influential authors. Among those psychiatrists responding to the survey (approximately 59%), some endorsed each definition of thought broadcast. CONCLUSIONS: Thought broadcast means different things to different people. Inconsistent terminology might impair communication in clinical and research contexts. PMID- 14519606 TI - Psychiatry in Britain one hundred years ago. PMID- 14519607 TI - Functional health status, chronic medical conditions and disorders of mood. AB - BACKGROUND: Understanding of the impact of depressive and anxiety disorders on functional health status in the context of chronic medical illness has been gained almost exclusively from the study of patient populations. AIMS: To compare the impact of major depressive and generalised anxiety disorder with that of chronic medical conditions on functional health in a UK resident population. METHOD: The functional health of 20,921 study participants was assessed using the Medical Outcomes Study Short Form 36 questionnaire. Depressive and anxiety disorder episode histories and chronic medical conditions were assessed using independent self-completed questionnaires. RESULTS: The degree of physical functional impairment associated with mood disorders was of equivalent magnitude to that associated with the presence of chronic medical conditions or with being some 12 years older. CONCLUSIONS: Depressive and anxiety disorders have a profound impact on functional health that is independent of chronic medical illness. Chronic anxiety is associated with physical health limitations in excess of those associated with chronic depression or any of the physical health conditions considered, except for stroke. PMID- 14519608 TI - Comorbidity of substance misuse and mental illness in community mental health and substance misuse services. AB - BACKGROUND: Improved management of mental illness and substance misuse comorbidity is a National Health Service priority, but little is known about its prevalence and current management. AIMS: To measure the prevalence of comorbidity among patients of community mental health teams (CMHTs) and substance misuse services, and to assess the potential for joint management. METHOD: Cross sectional prevalence survey in four urban UK centres. RESULTS: Of CMHT patients, 44% (95% CI 38.1-49.9) reported past-year problem drug use and/or harmful alcohol use; 75% (95% CI 68.2-80.2) of drug service and 85% of alcohol service patients (95% CI 74.2-93.1) had a past-year psychiatric disorder. Most comorbidity patients appear ineligible for cross-referral between services. Large proportions are not identified by services and receive no specialist intervention. CONCLUSIONS: Comorbidity is highly prevalent in CMHT, drug and alcohol treatment populations, but may be difficult to manage by cross-referral psychiatric and substance misuse services as currently configured and resourced. PMID- 14519609 TI - Stability of recall of military hazards over time. Evidence from the Persian Gulf War of 1991. AB - BACKGROUND: War time traumatic events are related to subsequent psychological and physical health, but quantifying the association is problematic. Memory changes over time and is influenced by psychological status. AIMS: To use a large, two stage cohort study of members of the UK armed forces to study changes in recall of both traumatic and 'toxic' hazards. METHOD: A questionnaire-based follow-up study assessed 2370 UK military personnel, repeating earlier questions about exposure to military hazards. RESULTS: The k statistics for reporting of hazards were good for some exposures, but very low for others. Gulf veterans reported more exposures over time (no significant rise in the Bosnia cohort). In the Gulf cohort only, reporting new exposures was associated with worsening health perception, and forgetting previously reported exposures with improved perception. We found no association between physical health, psychological morbidity or post-traumatic stress disorder symptoms and endorsement or non endorsement of exposures. CONCLUSIONS: Reporting of military hazards after a conflict is not static, and is associated with current self-rated perception of health. Self-report of exposures associated with media publicity needs to be treated with caution. PMID- 14519610 TI - Estimating psychological treatment effects from a randomised controlled trial with both non-compliance and loss to follow-up. AB - BACKGROUND: The Outcomes of Depression International Network (ODIN) trial evaluated the effect of two psychological interventions for the treatment of depression in primary care. Only about half of the patients in the treatment arm complied with the offer of treatment, prompting the question:'what was the effect of treatment in those patients who actually received it?' AIMS: To illustrate the estimation of the effect of receipt of treatment in a randomised controlled trial subject to non-compliance and loss to follow-up. METHOD: We estimated the complier average causal effect (CACE) of treatment. RESULTS: In the ODIN trial the effect of receipt of psychological intervention (an average of about 4 points on the Beck Depression Inventory) is about twice that of offering it. CONCLUSIONS: The statistical analysis of the results of a clinical trial subject to non-compliance to allocated treatment is now reasonably straightforward through estimation of a CACE and investigators should be encouraged to present the results of analyses of this type as a routine component of a trial report. PMID- 14519611 TI - Abusive experiences and psychiatric morbidity in women primary care attenders. AB - BACKGROUND: Abusive experiences in childhood and adulthood increase risks of psychiatric morbidity in women and independently increase risks of further abuse over the lifetime. It is unclear which experiences are most damaging. AIMS: To measure lifetime prevalence of abusive experiences and psychiatric morbidity, and to analyse associations in women primary care attenders. METHOD: A cross sectional, self-report survey of 1207 women attending 13 surgeries in the London borough of Hackney, UK. Independent associations between demographic measures, abusive experiences and psychiatric outcome were established using logistic regression. RESULTS: Childhood sexual abuse had few associations with adult mental health measures, in contrast to physical abuse. Sexual assault in adulthood was associated with substance misuse; rape with anxiety, depression and post-traumatic stress disorder but not substance misuse. Domestic violence showed strongest associations with most mental health measures, increased for experiences in the past year. CONCLUSIONS: Abuse in childhood and adulthood have differential effects on mental health; effects are increased by recency and severity. Women should be routinely questioned about ongoing and recent experiences as well as childhood. PMID- 14519613 TI - Maternal depression and child behaviour problems. Randomised placebo-controlled trial of a cognitive-behavioural group intervention. AB - BACKGROUND: Despite the frequently reported association between maternal depression and childhood psychopathological disorder, few studies have attempted to intervene with both conditions. AIMS: To evaluate the effect of group cognitive-behavioural therapy (CBT) on child behaviour problems and maternal depression in a group of women with young children. METHOD: An assessor-masked, randomised placebo-controlled trial compared three treatments: CBT for depression and parenting skills enhancement; a mothers' support group; and no intervention. An epidemiological (general population) sample was recruited. RESULTS: Analysis showed no significant difference between the groups. Within-group comparison suggested that at the end of treatment and at 6-month and 12-month follow-up, child problems and maternal depression had improved significantly in the CBT group. CONCLUSIONS: There was no statistically significant difference between groups. Both contact interventions seemed to provide some benefits to mothers with depression, with a possibly improved outcome resulting from CBT for children with behavioural problems. The results must be treated with caution. PMID- 14519614 TI - Common mental disorders among those attending primary health clinics and traditional healers in urban Tanzania. AB - BACKGROUND: Traditional healers provide a popular and accessible service across the African continent. Little is known of the characteristics or mental health status of those using these services. AIMS: To determine and compare the prevalence of common mental disorder among, and the characteristics of, those attending primary health care clinics (PHCs) and traditional healer centres (THCs) in Dar-es-Salaam. METHOD: The Clinical Interview Schedule-Revised was used to determine the prevalence of mental disorders in 178 patients from PHCs and 176 from THCs, aged 16-65 years. RESULTS: The prevalence of common mental disorders among THC patients (48%) was double that of PHC patients (24%). Being older, Christian, better educated, and divorced, separated or widowed were independently associated with THC attendance. None of these factors explained the excess of mental disorder among THC attenders. CONCLUSIONS: The high prevalence of mental disorders among THC attenders may reflect the failure of primary health care services adequately to detect and treat these common and disabling disorders. Traditional healers should be involved in planning comprehensive mental health care. PMID- 14519615 TI - Psychosocial treatment for severe personality disorder. 36-month follow-up. AB - BACKGROUND: In a previous report a step-down psychosocial programme for severe personality disorders was found to be more effective at expected termination of treatment than a longer in-patient treatment with no planned after-care. AIMS: To evaluate the clinical effectiveness of these two psychosocial specialist programmes over a 3-year follow-up period. METHOD: Two samples allocated to the in-patient treatment and to the step-down programme were compared prospectively on symptom severity, social adjustment, global assessment of mental health and other clinical indicators at 6, 12, 24 and 36 months after intake. RESULTS: Improvements were significantly greater in the step-down programme for social adjustment and global assessment of mental health. Patients in the programme were found to self-mutilate, attempt suicide and be readmitted significantly less at 24- and 36-month follow-up than patients in the in-patient group. CONCLUSIONS: Improvements associated with specialist residential treatment continued 2 years after discharge. A step-down model has significant advantages over a purely in patient model. PMID- 14519616 TI - Slovenia: difficulties and strengths of psychiatric research in a small country. PMID- 14519617 TI - Prophylaxis of depression in older people. PMID- 14519618 TI - Liaison psychiatry and general hospital management. PMID- 14519619 TI - Adverse events following neurosurgery. PMID- 14519621 TI - Neuroscience and psychodynamics. PMID- 14519620 TI - Liaison psychiatry and older adults. PMID- 14519623 TI - Recombinant human angiostatin by twice-daily subcutaneous injection in advanced cancer: a pharmacokinetic and long-term safety study. AB - PURPOSE: A clinical study was performed to evaluate the pharmacokinetics (PK) and toxicity of three dose levels of the angiogenesis inhibitor recombinant human (rh) angiostatin when administered twice daily by s.c. injection. EXPERIMENTAL DESIGN: Eligible patients had cancer not amenable to standard treatments. Three groups of 8 patients received 7.5, 15, or 30 mg/m(2)/day divided in two s.c. injections for 28 consecutive days followed by a 7-day washout period. PK assessment was done at days 1 and 28. Thereafter, in absence of toxicity or a 100% increase in tumor size, treatment was continued without interruption. RESULTS: Median age was 53 years (range, 43-75), male:female ratio 10:14, Eastern Cooperative Oncology Group performance 0-1. At the range of doses evaluated, serum PK of all 24 of the patients showed linear relation between dose and area under the curve (0- infinity) and C(max) (reached after 2 h). Thirteen of 24 patients developed erythema at injection sites (11 patients, CTC grade 1; 2 patients, CTC grade 2) without pain or itching, spontaneously resolving within 2 3 weeks of treatment. Two patients went off study after developing hemorrhage in brain metastases, and 2 patients developed deep venous thrombosis. No other relevant treatment-related toxicities were seen, even during prolonged treatment. A panel of coagulation parameters was not influenced by rhAngiostatin treatment. Long-term (>6 months) stable disease (<25% growth of measurable uni- or bidimensional tumor size) was observed in 6 of 24 patients. Five patients received rhAngiostatin treatment for >1 year (overall median time on treatment 99 days). CONCLUSIONS: Long-term twice-daily s.c. treatment with rhAngiostatin is well tolerated and feasible at the selected doses, and merits additional evaluation. Systemic exposure to rhAngiostatin is within the range of drug exposure that has biological activity in preclinical models. PMID- 14519624 TI - The androgen receptor gene is preferentially hypermethylated in follicular non Hodgkin's lymphomas. AB - This investigation examined promoter DNA methylation of the androgen receptor (AR) gene in non-Hodgkin's lymphoma (NHL) representing different stages of B-cell differentiation. Steroid hormones are important endocrine messengers with a broad range of physiological functions, including regulation of B-cell lymphopoiesis. Some of these effects are mediated via specific receptors such as AR that can act as a ligand-dependent transcription factor for other genes. DNA was isolated from 76 NHL specimens representing pregerminal center, germinal center, and postgerminal center states of differentiation. Initial methylation data were obtained from oligonucleotide microarrays and was confirmed and extended using methylation-specific PCR. Methylation of the AR gene promoter was present in a nonrandom pattern. Those tumors derived from pregerminal center or postgerminal center stages showed virtually no methylation and expressed AR mRNA. Cases of germinal center origin, mainly follicular lymphomas and some diffuse large B-cell lymphomas, showed hypermethylation. Studies with NHL cell lines revealed that demethylation or reversal of histone deacetylation partially restored AR expression but reversal of both simultaneously provided a synergistic release from suppression. Promoter methylation of AR occurs in a differentiation stage selective manner; those cases arising in the germinal center are preferentially methylated. Full re-expression of AR requires both demethylation and reacetylation, a finding that may affect treatment decisions. PMID- 14519625 TI - Association of tapasin and HLA class I antigen down-regulation in primary maxillary sinus squamous cell carcinoma lesions with reduced survival of patients. AB - PURPOSE: The purpose of this research was to assess the frequency and clinical significance of antigen processing machinery component and HLA class I antigen down-regulation in primary maxillary sinus squamous cell carcinoma (SCC) lesions. EXPERIMENTAL DESIGN: Formalin-fixed, paraffin-embedded tumor biopsy specimens at pretreatment status from 70 Japanese patients with maxillary sinus SCC were examined for HLA class I antigen and endoplasmic reticulum chaperone molecule expression using an immunohistochemical method. Furthermore, the results of immunohistochemical staining of the lesions were correlated with their histopathological characteristics and with the clinical course of the disease. RESULTS: Calnexin, ERp57, calreticulin, tapasin, and HLA class I antigens were down-regulated in 13, 13, 24, 69, and 78% of the 70 lesions tested, respectively. Both tapasin and HLA class I antigen expression were significantly correlated with the number of infiltrating CD3(+) T cells into tumor lesions (P < 0.01); furthermore, tapasin expression was significantly correlated with tumor differentiation (P = 0.024). Tapasin expression was correlated with that of HLA class I antigens (P < 0.01). Furthermore, tapasin and HLA class I antigen down regulation in SCC lesions was significantly associated with reduced survival of patients (P = 0.01 and P = 0.002, respectively). Multivariate Cox proportional hazards model analysis identified HLA class I antigen down-regulation as an independent prognostic marker. CONCLUSIONS: Tapasin expression appears to be associated with HLA class I antigen expression in primary maxillary sinus SCC lesions. Furthermore, defects in tapasin and HLA class I antigen expression in primary maxillary sinus SCC lesions may play a role in the clinical course of the disease, because these defects were associated with poor prognosis. PMID- 14519626 TI - All-trans-retinoic acid-induced apoptosis in human medulloblastoma: activation of caspase-3/poly(ADP-ribose) polymerase 1 pathway. AB - Current treatments for childhood brain tumor medulloblastoma (MB), radiation and chemotherapy, lead to undesirable side effects. Identification of antitumor agents that reduce the toxicity will thus have significant therapeutic value. In this study, we investigated all-trans-retinoic acid (ATRA) as an antitumor agent. Although high concentrations (1-10 microM) of retinoic acid derivatives are generally needed for significant antitumor effects in many cancer cells, we observed that pharmacologically relevant concentrations of ATRA were effective in inducing cell death in human MB cells. Using 10-fold lower concentrations (100 500 nM), we found that ATRA inhibits MB (DAOY, D283, D425, and D458) cell proliferation as determined by cell viability [3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide] and bromodeoxyuridine incorporation assays. Furthermore, 100 nM ATRA was potent in inhibiting the anchorage-independent growth of the sensitive cell lines (D283, D425, and D458) in soft agar assays. We also demonstrate that the ATRA-induced decrease in cell viability was due to increased cell death by apoptosis, which was accompanied by a 20-fold induction of caspase-3 activity in the most sensitive cell line, D458. By contrast, induction of caspase-3 was only 2-fold in the relatively insensitive DAOY cells. Furthermore, ATRA-induced cell death in D283, D425, and D458 cells was accompanied by activation of caspase-3, a key executioner of apoptosis. We also demonstrate that activated caspase-3 resulted in cleavage of 116-kDa poly(ADP ribose) polymerase 1 to its signature fragments (85 and 29 kDa). Pretreatment with a specific caspase-3 inhibitor, DEVD-CHO, significantly reduced ATRA-induced apoptotic cell death. Thus, we demonstrate for the first time that low concentrations of ATRA inhibit MB cell proliferation and induce apoptotic cell death in part by activating caspase-3/poly(ADP-ribose) polymerase 1 effector pathway, and we show that retinoic acids and novel retinoids are potential antitumor agents in MB therapy. PMID- 14519627 TI - The Bcl-2 transgene protects T cells from renal cell carcinoma-mediated apoptosis. AB - PURPOSE: Tumors induce T-cell apoptosis as a mechanism of inhibiting antitumor immunity. Using coculture experiments, it has been shown that tumor lines stimulate T-cell apoptosis by a pathway involving a mitochondrial permeability transition and cytochrome c release. Activated T cells express abundant levels of Bcl-2, an antiapoptotic molecule that would be expected to confer resistance to such tumor-mediated killing. We examined the mechanism by which Bcl-2 is dysregulated in T cells exposed to the renal tumor line SK-RC-45, and we determined whether overexpressing Bcl-2 protects T cells from tumor-mediated apoptosis. EXPERIMENTAL DESIGN: Activated T lymphocytes and Jurkat cells transfected or not transfected with Bcl-2 were exposed to SK-RC-45 for 48-72 h. After coculture, lymphocytes were analyzed for Bcl-2 expression using Western analysis and for tumor-induced apoptosis by terminal deoxynucleotidyl transferase mediated nick end labeling. The role of SK-RC-45-stimulated caspase activation in degrading T-cell Bcl-2 was assessed using a pan-caspase inhibitor, as well as a specific inhibitor of caspase-9. RESULTS: The renal cell carcinoma cell line SK RC-45 sensitizes peripheral blood activated T lymphocytes and Jurkat cells to apoptosis by a mechanism that involves degradation of the antiapoptotic protein Bcl-2. The SK-RC-45-induced modulation of lymphocyte Bcl-2 levels was largely caspase independent because pretreatment of T cells with pan-caspase inhibitor III or an inhibitor of caspase-9 had minimal or no effect on stabilizing the protein, although it did provide protection against apoptosis. Overexpression of Bcl-2 protected Jurkat cells from tumor-mediated killing. CONCLUSIONS: Bcl-2 inhibition is a mechanism by which tumors may render lymphocytes sensitive to other tumor-derived, proapoptotic stimuli. PMID- 14519628 TI - A safety study of a mixed-backbone oligonucleotide (GEM231) targeting the type I regulatory subunit alpha of protein kinase A using a continuous infusion schedule in patients with refractory solid tumors. AB - PURPOSE: The purpose of this study was to define the safety and pharmacodynamics of GEM231, a mixed backbone antisense oligonucleotide targeting the type I regulatory subunit alpha of protein kinase A, administered as a continuous i.v. infusion. EXPERIMENTAL DESIGN: Fourteen cancer patients received escalating doses of GEM231 as a 3-day (1 patient) or a 5-day continuous i.v. infusion (13 patients) at doses ranging from 80 to 180 mg/m(2)/day. RESULTS: The maximum tolerated dose of GEM231 was 180 mg/m(2)/day, based on dose-limiting elevation of serum transaminases (STs). At the recommended Phase II dose, 120 mg/m(2)/day (n = 8), the median number of cycles delivered was 2 (range, 1-4 cycles). Toxicities were tolerable, with one patient experiencing grade 3 ST elevation after 8 weeks. Plasma activated partial thromboplastin time changes were transient, reached a peak at the end of each weekly infusion, and were not associated with spontaneous bleeding. There was a significant difference between the mean preinfusion and postinfusion activated partial thromboplastin time measurements (2.05 s; P = 0.029). The most significant nonhematological toxicity was elevation in ST, usually observed after >/==" BORDER="0">4 weeks of therapy. There was a positive correlation between weekly dose and change in aspartate and alanine aminotransferase from baseline [r(2) = 0.56 (P = 0.031) and r(2) = 0.64 (P = 0.019), respectively]. ST elevations were reversible to near baseline in all patients within 3-4 weeks of interruption of GEM231 dosing. Low-grade fatigue was common (57%), cumulative by weeks 4-6, and reversible after GEM231 discontinuation. CONCLUSIONS: GEM231 administered as a continuous infusion is safe; however, continuous protracted dosing is limited by ST elevations. Alternative dosing schedules should include intermittent administration to minimize cumulative toxicity. Additional studies using intermittent continuous infusion schedules of GEM231 are warranted. PMID- 14519629 TI - Phase II study of 1alpha-hydroxyvitamin D(2) in the treatment of advanced androgen-independent prostate cancer. AB - PURPOSE: In this single institution Phase II trial, we evaluated the efficacy of the vitamin D analogue, 1alpha-OH-D(2), in patients with advanced hormone refractory prostate cancer. EXPERIMENTAL DESIGN: The patients initially received 1alpha-OH-D(2) at 12.5 micro g p.o. every day, which was dose adjusted for hypercalcemia. Given the cytostatic nature of the drug, the primary study end point was progression-free survival for a minimum of 6 months. The secondary end point was further characterization of drug toxicity. RESULTS: A total of 26 patients was enrolled. Using the intent-to-treat population, stable disease was seen for an average of 19.2 weeks (median 12 weeks, range 3-108 weeks). Twenty patients were evaluable for response. The one patient that achieved disease stabilization for >2 years elected to come off-study because of patient preference. His last disease evaluation showed no evidence of progression. No objective responses were seen. Previous and ongoing clinical observations strongly imply that PSA could be a misleading surrogate marker for clinical effect with this type of drug. Therefore, prostate-specific antigen was not used as a marker for disease response. Toxicity was as expected with mild hypercalcemia and associated symptoms like constipation and prerenal azotemia seen in some patients. Six (30%) evaluable patients experienced stable disease for >6 months, suggesting possible cytostatic activity. CONCLUSION: The results of this and other trials suggest further clinical investigation in this disease with vitamin D analogues alone or in combination with other agents, such as chemotherapy, should be pursued. PMID- 14519630 TI - Phase I study of oral topotecan in hematological malignancies. AB - PURPOSE: In this Phase I, dose-seeking study, we investigated the dose-limiting toxicities (DLTs) and maximal tolerated dose (MTD) of oral topotecan in patients with hematological malignancies. EXPERIMENTAL DESIGN: Patients with myelodysplastic syndromes, myeloproliferative disorders, or relapsed acute myelogenous leukemia were treated with 0.6-1.9 mg/m(2)/day oral topotecan for 5 consecutive days on and 2 days off, for 3 weeks (15 doses/course) followed by 2-4 weeks of rest. The DLTs occurring during the first course of treatment were considered for defining the MTD. Preliminary results of antitumor activity were assessed by examining bone marrow status and peripheral blood cell counts. RESULTS: All 26 patients enrolled in the study were evaluable for toxicity, and 24 patients were evaluable for response. A total of 54 courses were administered. The most frequently reported nonhematological toxicities (percentage of courses) were diarrhea (57%), nausea/vomiting (50%), fatigue (24%), and mucositis (9%). DLTs included grade 3 or 4 nausea/vomiting and diarrhea at 1.9 mg/m(2)/day. The MTD for oral topotecan in patients with hematological malignancies was defined at 1.4 mg/m(2)/day. Hematological toxicity was noted in all 26 patients and with all courses but was not considered dose-limiting. Four (17%) patients achieved a complete response, and six (25%) patients experienced hematological improvement. CONCLUSIONS: Protracted administration of oral topotecan is safe and well tolerated in patients with hematological malignancies. At the dose-schedule used, single-agent oral topotecan has a definite activity in patients with myelodysplastic syndrome and acute myelogenous leukemia and warrants further investigation alone or in combination with other agents. PMID- 14519631 TI - Phase I and pharmacokinetic study of triapine, a potent ribonucleotide reductase inhibitor, administered daily for five days in patients with advanced solid tumors. AB - PURPOSE: A Phase I study in patients with advanced cancer was conducted to determine the safety, pharmacokinetics, and maximum tolerated dose of Triapine, a new, potent small-molecule inhibitor of ribonucleotide reductase. EXPERIMENTAL DESIGN: Triapine was administered by 2-h i.v. infusion daily for 5 days. Courses were repeated every 4 weeks. The starting dose was 5 mg/m(2)/day, but was reduced to 2 mg/m(2)/day after the first patient developed a hepatic adverse event. The dose was subsequently escalated using a modified Fibonacci scheme in cohorts of 3 6 patients. After the 12 mg/m(2)/day dose level, the study design was amended to permit 100% dose escalation in single-patient cohorts until the first episode of a drug-related grade 2 adverse event or dose-limiting toxicity (DLT). On reaching a dose of 96 mg/m(2)/day, the study was amended to determine the safety and tolerability of the 96-mg/m(2) dose administered daily for 5 days every 2 weeks in an expanded cohort of patients. RESULTS: A total of 32 patients received treatment. During the dose escalation phase of the study, grade 2-4 drug-related adverse events were first observed at a dose of 96 mg/m(2)/day. Grade 3-4 leukopenia was the primary toxicity observed among four patients treated at this dose, which occurred in the week after treatment and resolved to grade 1 or lower by day 15. Fifteen patients were subsequently treated at the 96-mg/m(2) dose, daily for 5 days, with courses repeated every 2 weeks. The most common nonhematological toxicities for the latter schedule were asthenia, fever, nausea and vomiting, mucositis, decreased serum bicarbonate, and hyperbilirubinemia, and were predominantly grade 1-2 in severity and rapidly reversible. Hematological toxicity on the every-other-week schedule consisted of leukopenia (grade 4 in 93% in at least one course) and anemia (grade 2 in 71%, grade 3 in 22%). Thrombocytopenia was less common and was grade 3-4 in severity in only 22%. Triapine showed linear pharmacokinetic behavior although interpatient variability was relatively high. Peak concentrations at the 96-mg/m(2)/day dose averaged 8 microM, and the mean elimination T(1/2) ranged from 35 min to 3 h, with a median value of approximately 1 h. Cumulative urinary recovery averaged 1-3% of the administered dose, suggesting that the elimination of Triapine was primarily through metabolism. No partial or complete responses were observed. CONCLUSIONS: Triapine administered at a dose of 96 mg/m(2) by 2-h i.v. infusion daily for 5 days on an every-other-week schedule demonstrates an acceptable safety profile. Serum concentrations that surpass in vitro tumor growth-inhibitory concentrations are achieved for brief periods of time each day and are sufficient to produce myelosuppression, the expected consequence of ribonucleotide reductase inhibition. Phase II trials are indicated but will proceed with a daily-for-4 days schedule to reduce the incidence of grade 4 leukopenia. The safety profile also supports the initiation of Phase I combination trials with other anticancer agents. PMID- 14519632 TI - Phase I pharmacological and bioavailability study of oral diflomotecan (BN80915), a novel E-ring-modified camptothecin analogue in adults with solid tumors. AB - PURPOSE: Diflomotecan (BN80915) is an E-ring modified camptothecin analogue that possesses greater lactone stability in plasma compared with other topoisomerase I inhibitors, a potential advantage for antitumor activity. As with other camptothecins, oral administration has pharmacological and clinical advantages. This Phase I study was performed to assess the feasibility of the administration of oral diflomotecan, to determine the maximum-tolerated, dose its bioavailability, and to explore the pharmacokinetics. EXPERIMENTAL DESIGN: An initial i.v. bolus was administered to assess the bioavailability of diflomotecan. Fourteen days later, diflomotecan was administered p.o. once daily for 5 days to adult patients with solid malignant tumors and repeated every 3 weeks. BN80915 and its open lactone form BN80942 were measured. RESULTS: Twenty two patients entered the study and received a total of 57 cycles of oral diflomotecan at flat dose levels of 0.10, 0.20, 0.27, and 0.35 mg. The main toxicity was hematological, but some patients experienced alopecia, mild gastrointestinal toxicity, and fatigue. At the 0.35-mg dose level, 2 of 4 patients experienced dose-limiting toxicity comprising grade 3 thrombocytopenia with epistaxis and febrile neutropenia in 1 patient and uncomplicated grade 4 neutropenia lasting for >7 days in another. Toxicity was acceptable at the 0.27 mg dose level at which dose-limiting toxicities were observed in 3 of 12 patients (grade 4 neutropenia > 7 days, complicated by fever in 1 patient but without other signs of infection). After two cycles of diflomotecan, 6 patients had disease stabilization, which was maintained in 2 patients for 9 months and >1 year, respectively. Diflomotecan pharmacokinetics were linear over the dose range studied. Systemic exposure correlated with the fall in WBC counts. The mean oral bioavailability (+/-SD) was 72.24 +/- 59.2% across all dose levels. Urinary excretion of BN80915 was very low. CONCLUSIONS: The recommended oral diflomotecan dose for Phase II studies is 0.27 mg/day x 5 every 3 weeks. This regimen is convenient and generally well tolerated with a favorable pharmacokinetic profile and high but variable bioavailability. PMID- 14519633 TI - A phase I/IIA trial of continuous five-day infusion of squalamine lactate (MSI 1256F) plus carboplatin and paclitaxel in patients with advanced non-small cell lung cancer. AB - PURPOSE: Squalamine is an antitumor agent that has been shown to have antiangiogenic activity in animal models. This Phase I/IIA study was designed to assess the safety, clinical response, and pharmacokinetics of squalamine when administered as a 5-day continuous infusion in conjunction with standard chemotherapy every 3 weeks in patients with stage IIIB (pleural effusion) or stage IV non-small cell lung cancer. EXPERIMENTAL DESIGN: Patients with chemotherapy-naive non-small cell lung cancer were treated with escalating doses of squalamine in combination with standard doses of paclitaxel and carboplatin. Paclitaxel and carboplatin were administered on day 1, followed by squalamine as a continuous infusion on days 1-5, every 21 days. RESULTS: A total of 45 patients were enrolled (18 patients in the Phase I dose escalation arm and 27 in the Phase IIA arm). The starting dose of squalamine was 100 mg/m(2)/day and escalated to 400 mg/m(2)/day; two of three patients at 400 mg/m(2)/day had dose-limiting toxicity that included grade 3/4 arthralgia, myalgia, and neutropenia. On the basis of safety and toxicity, 300 mg/m(2)/day was selected as the Phase II dose of squalamine in this combination regimen. An additional 27 patients (a total of 33) were enrolled according to the protocol treatment schema at 300 mg/m(2)/day. There was no pharmacokinetic evidence of drug interactions for the combination of squalamine, carboplatin, and paclitaxel. Forty-three patients were evaluable for response. Partial tumor responses were observed in 12 (28%) of these patients; an additional 8 evaluable patients (19%) were reported to have stable disease. For all of the patients treated, the median survival was 10.0 months; and 1-year survival was 40%. CONCLUSIONS: The combination of squalamine given continuously daily for 5 days, with paclitaxel and carboplatin given on day 1, is well tolerated. Patient survival data and the safety profile of this drug combination suggests that the use of squalamine given at its maximum tolerated dose with cytotoxic chemotherapy should be explored further as a potentially effective therapeutic strategy for patients with stage IIIB or IV non-small cell lung cancer. PMID- 14519634 TI - Thymidylate synthase and dihydropyrimidine dehydrogenase mRNA expression levels: predictors for survival in colorectal cancer patients receiving adjuvant 5 fluorouracil. AB - PURPOSE: Despite adjuvant 5-fluorouracil (5-FU) therapy, approximately 30% of patients with International Union against Cancer stage II and III colorectal cancer develop recurrence. In this study, we determined the prognostic value of thymidylate synthase (TS) and dihydropyrimidine dehydrogenase (DPD) expression in colorectal cancer patients treated with adjuvant 5-FU. EXPERIMENTAL DESIGN: A real-time reverse transcription-PCR technique for quantitation of relative gene expression from paraffin-embedded specimen was established first. In a second step, archival paraffin-embedded primary tumor tissue of 309 patients who participated in adjuvant colorectal cancer trials was analyzed for TS and DPD mRNA expression. RESULTS: TS mRNA expression determined by real-time reverse transcription-PCR correlated with TS protein levels determined by TS immunoblot and immunohistochemistry in cultured colon cancer cell lines and paraffin embedded primary tumor tissue. TS mRNA levels in fresh-frozen tissues also correlated with TS mRNA levels in corresponding paraffin sections. Among the patients receiving adjuvant 5-FU therapy, those with high TS survived longer than those with low TS, and in each TS subgroup, the ones with low DPD survived longer than the ones with high DPD levels. Multiple Cox regression analysis showed that besides tumor stage (P = 0.010), only the combination of TS and DPD expression turned out to be an independent prognostic factor for survival (P = 0.030). CONCLUSIONS: This suggests that TS and DPD quantitation may be helpful to evaluate prognosis of patients receiving adjuvant 5-FU and that patients with high TS and low DPD may benefit from adjuvant 5-FU chemotherapy. PMID- 14519635 TI - Expression of endothelin-1, endothelin-A, and endothelin-B receptor in human breast cancer and correlation with long-term follow-up. AB - PURPOSE: Endothelin-1 (ET-1) is overexpressed in breast carcinomas and stimulates tumor cell growth in an autocrine and paracrine fashion via its receptors, ET(A)R and ET(B)R. In this study, we evaluated the expression of ET-1 and ET receptors in breast carcinomas and determined its clinical and prognostic significance. EXPERIMENTAL DESIGN: We analyzed expression of ET-1, ET(A)R, and ET(B)R in 176 breast carcinomas using a semiquantitative immunohistochemical approach. Statistical analysis of clinicopathological variables such as pT stage, pN stage, hormone receptor status, Her-2/neu amplification, histological grade, and long term follow-up data were performed. RESULTS: We observed a moderate to strong cytoplasmic staining for ET-1 in 69 (43.1%), for ET(A)R in 74 (46.5%), and for ET(B)R in 86 (53.4%) cases of primary breast cancer. A correlation was found between increased ET-1 expression and its receptors with several clinicopathological parameters that characterize aggressive types of breast cancer, with the exception of increased ET(A)R and ET(B)R expression with positive estrogen receptor status. Elevated expression of ET-1, ET(A)R, and ET(B)R was more common in breast carcinomas of patients with lower disease-free survival time and overall survival. In addition, a statistically significant correlation was observed between ET(A)R expression and reduced disease-free survival time (P = 0.041). Interestingly, the prognostic impact of ET(A)R expression was shown to be more pronounced in the subgroup of patients with a putative favorable prognosis according to classic prognostic factors. CONCLUSIONS: Therefore, analysis of ET(A)R expression may improve the prediction of relapse and death and facilitate an individually based risk-directed adjuvant therapy in breast cancer patients. PMID- 14519636 TI - p15INK4b, p14ARF, and p16INK4a inactivation in sporadic and neurofibromatosis type 1-related malignant peripheral nerve sheath tumors. AB - PURPOSE: Malignant peripheral nerve sheath tumor (MPNST) can arise sporadically or in association with neurofibromatosis type 1. Deletions at the 9p21 locus have been reported in these tumors. To additionally characterize the status of this chromosomal region, in this study we performed a comprehensive, mostly PCR-based molecular analysis of the three tumor suppressor genes p15(INK4b), p14(ARF) and p16(INK4a) located at the 9p21 locus in 26 cryopreserved MPNSTs. EXPERIMENTAL DESIGN: Fourteen neurofibromatosis type 1-related and 12 sporadic cases were investigated for homozygous deletion coupled with fluorescent in situ hybridization, promoter methylation, and mutational analysis, as well as m-RNA expression. RESULTS: The results showed that an inactivation of one or more genes occurred in 77% of MPNSTs and was mainly achieved through homozygous deletion (46%), which, in turn, encompassed all of the three tandemly linked genes in 83% of the deleted cases. Promoter methylation was at a less extent involved in gene silencing (18%), and no mutations were found. Loss of function at DNA level strongly correlated with loss of mRNA expression accounting for 80% of the cases. Because of the close relationship between p14(ARF) and TP53 and between p15(INK4b)/p16(INK4a) and Rb, these results support a model of a coinactivation of TP53 and Rb pathways in 75% of MPNSTs, with functional consequences on cell growth control and apoptosis. CONCLUSIONS: The inactivation of the 9p21 locus is a frequent and peculiar hallmark of MPNST genetic profile leading also to an impaired apoptosis that could be taken into account in treatment planning of these tumors. PMID- 14519637 TI - The impact of p53 protein core domain structural alteration on ovarian cancer survival. AB - PURPOSE: Although survival with a p53 missense mutation is highly variable, p53 null mutation is an independent adverse prognostic factor for advanced stage ovarian cancer. By evaluating ovarian cancer survival based upon a structure function analysis of the p53 protein, we tested the hypothesis that not all missense mutations are equivalent. EXPERIMENTAL DESIGN: The p53 gene was sequenced from 267 consecutive ovarian cancers. The effect of individual missense mutations on p53 structure was analyzed using the International Agency for Research on Cancer p53 Mutational Database, which specifies the effects of p53 mutations on p53 core domain structure. Mutations in the p53 core domain were classified as either explained or not explained in structural or functional terms by their predicted effects on protein folding, protein-DNA contacts, or mutation in highly conserved residues. Null mutations were classified by their mechanism of origin. RESULTS: Mutations were sequenced from 125 tumors. Effects of 62 of the 82 missense mutations (76%) could be explained by alterations in the p53 protein. Twenty-three (28%) of the explained mutations occurred in highly conserved regions of the p53 core protein. Twenty-two nonsense point mutations and 21 frameshift null mutations were sequenced. Survival was independent of missense mutation type and mechanism of null mutation. CONCLUSIONS: The hypothesis that not all missense mutations are equivalent is, therefore, rejected. Furthermore, p53 core domain structural alteration secondary to missense point mutation is not functionally equivalent to a p53-null mutation. The poor prognosis associated with p53-null mutation is independent of the mutation mechanism. PMID- 14519638 TI - Early stage cervical cancers containing human papillomavirus type 18 DNA have more nodal metastasis and deeper stromal invasion. AB - PURPOSE: To investigate the clinical and pathological factors which might explain the poor prognosis associated with early stage cervical cancers containing human papillomavirus (HPV) type 18 DNA. EXPERIMENTAL DESIGN: A clinical and pathological review of 144 patients with stage IB cervical cancer treated with radical hysterectomy and bilateral pelvic lymph node dissection was done. HPV genotyping was determined from fresh tumor specimens through PCR. Clinical pathological information, sites of recurrence, use of adjuvant radiation, and survival data were analyzed. RESULTS: Thirty-three (23%) tumors contained HPV 18 DNA. These tumors did not differ from those which contained non-HPV 18 DNA with respect to tumor grade or size. However, HPV 18-containing cancers were more likely to be adenocarcinomas. A higher incidence of pelvic lymph node metastasis was noted among the HPV 18 group (48%) as compared with the non-HPV 18 group (28%), and deeper stromal invasion was more common in HPV 18-associated tumors. Although a slightly higher proportion of patients with HPV 18-containing tumors received adjuvant radiation (67%) than those with non-HPV 18 cancers (49%), recurrences were more common among HPV 18 patients. Eleven (33%) of HPV 18 containing cancers relapsed compared with 18 (16%) of non-HPV18-containing tumors. CONCLUSIONS: The explanation for the worse prognosis associated with stage IB cervical cancers containing HPV 18 DNA treated with radical hysterectomy and bilateral pelvic lymph node dissection appears to be related to deeper cervical stromal invasion and more nodal metastases. Despite an increased use of adjuvant radiation therapy, these cancers are still more likely to relapse. PMID- 14519639 TI - Short postoperative survival for glioblastoma patients with a dysfunctional Rb1 pathway in combination with no wild-type PTEN. AB - PURPOSE: Glioblastoma (GB, WHO grade IV) is the most common primary brain tumor in adults. Survival is typically <1 year but varies between a few months and a couple of years. The aim of the study was to find novel genetic prognostic factors in a well-defined GB series. EXPERIMENTAL DESIGN: The survival data on 129 GBs were correlated with the results of a detailed analysis of 9 genes. These included 3 genes coding for proteins in the p53 pathway (i.e., TP53, p14(ARF), and MDM2), 4 in the Rb1 pathway (i.e., CDKN2A, CDKN2B, RB1, and CDK4), as well as PTEN and epidermal growth factor receptor. RESULTS: We found that abnormalities in any of the four genes (CDKN2A, CDKN2B, RB1, and CDK4) coding for components of the Rb1 pathway were associated with shorter survival (P = 0.002). In combination with loss of wild-type PTEN, the association was even stronger (P < 0.001), the median survival being 166 days as compared with the group without these abnormalities where the median postoperative survival was 437 days. The survival difference remained statistically significant in Cox' regression analysis adjusting for age (P = 0.012). CONCLUSIONS: The findings indicate that knowledge of the molecular genetic abnormalities in GBs provides important data in assessing individual patients. As additional advances in our understanding of the molecular genetics and cell biology of gliomas are made, in addition to providing prognostic information, such data may also provide targets for innovative therapy in the individual case. PMID- 14519640 TI - Major histocompatibility antigens and antigen-processing molecules in uveal melanoma. AB - PURPOSE: Malignant transformation of cells is frequently associated with abnormalities in the human leukocyte antigen (HLA) expression. These abnormalities may play a role in the clinical course of the disease, because HLA antigens mediate interactions of tumor cells with T cells and natural killer cells. Uveal melanoma is a highly malignant tumor of the eye and is characterized by hematogenic spread to liver. Antigen-processing molecules (APMs) are necessary for efficient expression of HLA class I antigens. We studied the expression of HLA antigens and the APM in uveal melanomas by immunohistochemistry and correlated clinicopathologically. EXPERIMENTAL DESIGN: HLA class I antigen, beta(2)-microglobulin (beta(2)-m), HLA class II antigens, and the APM comprising proteasomal subunits low molecular mass polypeptide (LMP) 2, beta-subunit of LMP2 Delta, LMP 10, transporter associated protein 1 subunit, and chaperone molecules tapasin and calnexin were studied in 41 primary uveal melanoma archival specimens by immunohistochemistry. Immunoanalysis was done by a semiquantitative method and correlated with extrascleral extension, cell types, and the largest tumor diameter. RESULTS: HLA class I antigen, beta(2)-m, HLA class II antigen, and the APM were decreased (negative staining in 29 tumors and dull staining in 3 tumors) in 100% (32 of 32) uveal melanomas with no extrascleral extension. (P = 0.01) and positive (bright staining) in 67% (4 of 9) tumors with liver metastasis. Decreased immunoexpression of HLA antigens and the APM was seen in nonepithelioid cell melanomas. There was no correlation with largest tumor diameter. CONCLUSIONS: Our data suggest decreased expression of HLA, and APM are seen in uveal melanomas with no extrascleral extension and in nonepithelioid cell melanomas. Decreased expression of APM may contribute to decreased HLA class I antigen expression. PMID- 14519641 TI - Thymidylate synthase expression predicts the response to 5-fluorouracil-based adjuvant therapy in pancreatic cancer. AB - PURPOSE: Thymidylate synthase (TS) is the target enzyme for 5-fluorouracil (5 FU), and TS expression may determine clinical response and survival after therapy with 5-FU in colorectal cancer. 5-FU is also widely used in the adjuvant therapy of pancreatic cancer. Therefore, we explored the hypothesis that TS expression was associated with patient prognosis and the response to adjuvant therapy in pancreatic cancer. EXPERIMENTAL DESIGN: Cylindrical tissue cores from a large retrospective, nonrandomized series covering 132 resected patients were used to build a pancreatic cancer tissue microarray. TS expression was determined using immunohistochemistry. RESULTS: High intratumoral TS expression and low intratumoral TS expression were present in 83 of 132 (63%) and 49 of 132 (37%) tumors, respectively. Median survival among patients with low intratumoral TS expression (18 months) was longer than that among patients with high TS expression (12 months). In multivariate analysis, more advanced pathological stage [risk ratio (RR) = 1.70; P = 0.015], poorly differentiated histology (RR = 1.71; P = 0.015), management with adjuvant therapy (RR = 0.49; P = 0.011), and high TS expression [RR = 1.66; 95% confidence interval (CI) = 1.05-2.63; P = 0.029] were independent predictors of mortality. The risk of death was significantly reduced by any adjuvant therapy (RR = 0.40; 95% CI = 0.18-0.90; P = 0.001) among patients with high TS expression. This difference in survival among patients with low- and high-TS-expressing tumors became more significant when the analysis was restricted to the 73 patients receiving 5-FU-based adjuvant therapy (RR = 0.37; 95% CI = 0.16-0.86; P = 0.0006). In contrast, 5-FU-based adjuvant therapy did not influence survival among patients with low-TS-expressing pancreatic cancer. CONCLUSIONS: High TS expression is a marker of poor prognosis in resected pancreatic cancer. Patients with high intratumoral TS expression benefit from adjuvant therapy. PMID- 14519642 TI - Artificial intelligence in predicting bladder cancer outcome: a comparison of neuro-fuzzy modeling and artificial neural networks. AB - PURPOSE: New techniques for the prediction of tumor behavior are needed, because statistical analysis has a poor accuracy and is not applicable to the individual. Artificial intelligence (AI) may provide these suitable methods. Whereas artificial neural networks (ANN), the best-studied form of AI, have been used successfully, its hidden networks remain an obstacle to its acceptance. Neuro fuzzy modeling (NFM), another AI method, has a transparent functional layer and is without many of the drawbacks of ANN. We have compared the predictive accuracies of NFM, ANN, and traditional statistical methods, for the behavior of bladder cancer. EXPERIMENTAL DESIGN: Experimental molecular biomarkers, including p53 and the mismatch repair proteins, and conventional clinicopathological data were studied in a cohort of 109 patients with bladder cancer. For all three of the methods, models were produced to predict the presence and timing of a tumor relapse. RESULTS: Both methods of AI predicted relapse with an accuracy ranging from 88% to 95%. This was superior to statistical methods (71-77%; P < 0.0006). NFM appeared better than ANN at predicting the timing of relapse (P = 0.073). CONCLUSIONS: The use of AI can accurately predict cancer behavior. NFM has a similar or superior predictive accuracy to ANN. However, unlike the impenetrable "black-box" of a neural network, the rules of NFM are transparent, enabling validation from clinical knowledge and the manipulation of input variables to allow exploratory predictions. This technique could be used widely in a variety of areas of medicine. PMID- 14519643 TI - Squamous cell carcinoma of the esophagus can be detected by microsatellite analysis in tumor and serum. AB - PURPOSE: Esophageal squamous cell cancer can be treated effectively by potentially curative surgery if diagnosed at an early stage. Our aim was to develop a novel molecular approach as a noninvasive test for squamous cell cancer detection and as an indicator for the prognosis of the patients. EXPERIMENTAL DESIGN: Matched normal, tumor, and serum samples were obtained from 28 patients with squamous cell carcinoma (SCC) of the esophagus. DNA was extracted, and the samples were subjected to microsatellite analysis using 12 markers. Serum and normal DNA from 10 healthy individuals served as controls. RESULTS: Twenty-six of the 28 patients (92.9%) with SCC were found to have one or more microsatellite DNA alterations in their primary tumor. Twenty-seven of the 28 patients (96.4%) had at least one alteration in the serum by microsatellite analysis. Mean age was 61.5 years. Microsatellite alterations were not identified in the serum DNA of samples from normal control subjects. Median follow-up was 13 months. Survival and recurrence were not significantly correlated with either loss of heterozygosity in the tumor or in the serum. CONCLUSIONS: Microsatellite DNA analysis of tumor and serum specimen is a potentially valuable tool for detection and for the evaluation of the prognosis of SCC of the esophagus. The follow-up in our study is still too short to draw final conclusions on the correlation of disease-specific survival and disease recurrence with microsatellite alterations. The evidence of circulating tumor DNA in almost all of our patients underlines a systemic component of the disease that is not surgically amenable. PMID- 14519644 TI - Two prognostic groups of inflammatory breast cancer have distinct genotypes. AB - PURPOSE: The prognosis of inflammatory breast cancer (IBC) remains poor despite the use of multimodality treatments, with a 10-year survival rate of not >30%. Clinicopathological and biological predictors of outcome are inadequate in this setting. Analysis of loss of heterozygosity (LOH) can provide a molecular portrait of the genetic alterations underlying stepwise cancer progression. We tested the value of LOH patterns as diagnostic and prognostic markers in IBC. EXPERIMENTAL DESIGN: In a previous study of 64 patients with IBC who were treated homogeneously between 1988 and 1999, we determined LOH frequencies at 71 loci located in 20 chromosomal regions associated with primary breast cancer. Six of these regions bore alterations that were less frequent in non-IBC. In the present study, we sought correlations between these molecular data and the clinicopathological features and clinical outcome of the same 64 patients. RESULTS: With the exception of stage IV disease, extensive breast inflammation at first clinical examination was the main factor associated with poor outcome (P = 0.00065 versus localized inflammation). The overall frequency of LOH was also higher in this group (P = 0.000073). LOH patterns differed between patients with localized and extensive breast inflammation. CONCLUSION: Patients with IBC can be separated into two major prognostic groups on the basis of initial clinical signs, which appear to be subtended by different molecular alterations. PMID- 14519645 TI - Consequences of loss of progesterone receptor expression in development of invasive endometrial cancer. AB - PURPOSE: In endometrial cancer, loss of progesterone receptors (PR) is associated with more advanced disease. This study aimed to investigate the mechanism of action of progesterone and the loss of its receptors (PRA and PRB) in development of endometrial cancer. EXPERIMENTAL DESIGN: A 9600-cDNA microarray analysis was performed to study regulation of gene expression in the human endometrial cancer subcell line Ishikawa PRAB-36 by the progestagen medroxy progesterone acetate (MPA). Five MPA-regulated genes were selected for additional investigation. Expression of these genes was studied by Northern blot and by immunohistochemistry in Ishikawa subcell lines expressing different PR isoforms. Additionally, endometrial cancer tissue samples were immunohistochemically stained to study the in vivo protein expression of the selected genes. RESULTS: In the PRAB-36 cell line, MPA was found to regulate the expression of a number of invasion- and metastasis-related genes. On additional investigation of five of these genes (CD44, CSPG/Versican, Tenascin-C, Fibronectin-1, and Integrin-beta 1), it was observed that expression and progesterone regulation of expression of these genes varied in subcell lines expressing different PR isoforms. Furthermore, in advanced endometrial cancer, it was shown that loss of expression of both PR and E-cadherin was associated with increased expression CD44 and CSPG/Versican. CONCLUSION: The present study shows that progestagens exert a modulatory effect on the expression of genes involved in tumor cell invasion. As a consequence, loss of PR expression in human endometrial cancer may lead to development of a more invasive phenotype of the respective tumor. PMID- 14519646 TI - The correlation between aberrant connexin 43 mRNA expression induced by promoter methylation and nodal micrometastasis in non-small cell lung cancer. AB - Reduced connexin (Cx) 43 gene expression has been shown in most of lung tumors and cancer cell lines. Although aberrant Cx43 gene expression was linked with lung tumorigenesis, our understanding to the mechanism was still limited. We hypothesized that the evidence of aberrant Cx43 gene expression was gradually intensified from adjacent normal lung tissues surrounding tumors toward tumor tissues. In this study, 90 lung tumors and adjacent normal tissues were collected to examine Cx43 mRNA expression by reverse transcription-PCR (RT-PCR). Our data showed that Cx43 mRNA expression in adjacent normal lung tissue was significantly correlated with nodal involvement (P = 0.03), but the similar trend was not observed in tumor tissues. To verify whether lack of Cx43 mRNA expression resulted from promoter methylation, PCR-based methylation assay was performed for Cx43 promoter methylation analysis. A higher frequency of promoter methylation was observed in Cx43 mRNA-negative patients (21 of 33, 63.7%) compared with Cx43 mRNA-positive patients (3 of 57, 5.3%, P < 0.0001). To elucidate whether aberrant Cx43 gene expression originated from adjacent normal lung tissues, 25 lung tumors and each of five adjacent normal tissues at various distances from tumor tissues were collected to examine Cx43 mRNA and protein expression by RT-PCR and Western blot, respectively. The results show that Cx43 mRNA and protein expressions gradually decreased from adjacent normal lung tissues to tumor tissues with a positive correlation to the distance from the tumor tissues. Gel-shift assay data also revealed that shifted band binding with AP1 was only observed in adjacent normal tissues, which were far from the tumor tissues. These results indicate that promoter methylation may interfere with AP1 binding to the promoter to cause aberrant Cx43 gene expression. Thus, Cx43 mRNA in adjacent normal tissue surrounding lung tumor simply detected by RT-PCR may act as a molecular marker of nodal micrometastasis in non-small cell lung cancer. PMID- 14519647 TI - Responsiveness to the retinoic acid receptor-selective retinoid LGD1550 correlates with abrogation of transforming growth factor alpha/epidermal growth factor receptor autocrine signaling in head and neck squamous carcinoma cells. AB - PURPOSE: The use of retinoids to prevent and/or treat cancers, including head and neck squamous cell carcinoma (HNSCC), has been limited by the development of resistance and unwanted side effects. Receptor-selective retinoids are potentially less toxic than nonselective compounds. The present investigation was undertaken to determine the mechanism of responsiveness to an retinoic acid receptor-selective retinoid (LGD1550) that has shown antitumor efficacy in a xenograft model of HNSCC. EXPERIMENTAL DESIGN: A series of HNSCC cell lines were characterized with respect to proliferation and apoptosis after LGD1550 treatment. Relative responsiveness to LGD1550 was examined with respect to modulation of epidermal growth factor receptor (EGFR) signaling pathways. RESULTS: Cells were either growth inhibited and underwent apoptosis or were resistant to treatment with this compound. Retinoids have been shown to decrease the gene transcription rates of transforming growth factor (TGF)-alpha and EGFR in HNSCC. LGD1550 responsiveness was accompanied by decreased expression of TGF alpha, EGFR, and modulation of EGFR signaling pathways, including signal transducers and activators of transcriptions and mitogen-activated protein kinase. In contrast, EGFR autocrine signaling pathways were not altered in HNSCC cells that were resistant to the growth inhibitory effects of LGD1550. CONCLUSIONS: These results suggest that there is a correlation between the efficacy of receptor-selective retinoids and modulation of TGF-alpha/EGFR signaling in HNSCC. Therefore, alterations of these signaling pathways may serve as a biomarker of clinical response. PMID- 14519648 TI - Platelet-activating factor inactivation by local expression of platelet activating factor acetyl-hydrolase modifies tumor vascularization and growth. AB - PURPOSE: Platelet-activating factor (PAF), a phospholipid mediator of inflammation, has been recently detected on tumor cells but its effect in tumor development is largely undefined. EXPERIMENTAL DESIGN: To address its potential role in tumor biology, we inhibited intratumor PAF activity by engineering tumor cell lines to express plasma PAF-acetylhydrolase (PAF-AH), the major PAF inactivating enzyme, and studied their behavior in vitro and in vivo. RESULTS: When transfected with PAF-AH, KS-Imm human Kaposi's sarcoma cells implanted in SCID mice and B16F10 mouse melanoma cells implanted in syngenic C57Bl/6J mice showed significantly reduced vascularization and growth allowing longer survival compared with control tumors. The amounts of bioactive PAF extracted from PAF-AH transfected tumors were significantly reduced. In vitro, expression of PAF-AH did not influence cell proliferation, whereas it inhibited PAF-dependent cell motility in Kaposi's sarcoma cells that express PAF-receptor but not in melanoma cells that did not express it. On the other hand, PAF-induced endothelial tubulogenesis in Matrigel was inhibited by incubation with supernatant from PAF AH-transfected melanoma cells, indicating that PAF-AH inhibits in vitro neoangiogenesis. CONCLUSIONS: We demonstrated that in situ PAF inactivation affects tumor vascularization and growth through inhibition of neoangiogenesis and, in the case of cells expressing PAF receptor, also tumor cell motility. PMID- 14519649 TI - Differential vascular and transcriptional responses to anti-vascular endothelial growth factor antibody in orthotopic human pancreatic cancer xenografts. AB - PURPOSE: The objectives of this study were to investigate the effects of anti vascular endothelial growth factor (VEGF) treatment on various vascular functions, gene expression, and growth of orthotopic human pancreatic cancer xenografts and thus to provide useful preclinical data for novel cancer treatments. EXPERIMENTAL DESIGN: Small pieces of a human pancreatic carcinoma, PANC-1, were implanted into the pancreas of male severe combined immunodeficient mice. The animals were treated with anti-human VEGF antibody A.4.6.1 (300 micro g, every 3 days i.p.) or a nonspecific IgG between 4 and 8 weeks after tumor implantation. Then, vascular density, diameter, permeability, and tumor growth were determined by intravital microscopy. Subsequently, tumors were harvested, and angiogenic gene expression profile was determined by a microarray kit including 96 genes involved in tumor angiogenesis. RESULTS: Anti-VEGF antibody significantly reduced angiogenesis and growth of orthotopic PANC-1 tumors. In the anti-VEGF treatment group, the vessel density was significantly smaller (67.8 +/- 10.6 cm/cm(2)) than that seen in the control group (146.7 +/- 10.0 cm/cm(2)). However, vessel diameter and permeability were not altered significantly by anti VEGF antibody treatment. The pancreatic tumors in the treated group were significantly smaller than those in the control group. Microarray and subsequent Northern blot and semiquantitative reverse transcription-PCR analyses revealed both a decrease (fibroblast growth factor 1, transforming growth factor beta1, platelet-derived growth factor alpha, erbB2, and c-ets1,) and an increase (placenta growth factor, hypoxia-inducible factor alpha, and endoglin) in expression of angiogenesis-related genes in the PANC-1 tumors by anti-VEGF treatment. CONCLUSIONS: Anti-VEGF antibody treatment has differential effects on vessel functions as well as angiogenic gene expression and inhibitory effects on angiogenesis and growth of the orthotopic pancreatic tumor. Anti-VEGF strategy appears promising for pancreatic cancer treatment. PMID- 14519650 TI - Clinical predictive value of the in vitro cell line, human xenograft, and mouse allograft preclinical cancer models. AB - PURPOSE: We looked at the value of three preclinical cancer models, the in vitro human cell line, the human xenograft, and the murine allograft, to examine whether they are reliable in predicting clinical utility. EXPERIMENTAL DESIGN: Thirty-one cytotoxic cancer drugs were selected. Literature was searched for drug activity in Phase II trials, human xenograft, and mouse allografts in breast, non small cell lung, ovary, and colon cancers. Data from the National Cancer Institute Human Tumor Cell Line Screen were used to calculate drug in vitro preclinical activity for each cancer type. Phase II activity versus preclinical activity scatter plot and correlation analysis was conducted for each model, by tumor type (disease-oriented approach), using one tumor type as a predictor of overall activity in the other three tumor types combined (compound-oriented approach) and for all four tumor types together. RESULTS: The in vitro cell line model was predictive for non-small cell lung cancer under the disease-oriented approach, for breast and ovarian cancers under the compound-oriented approach, and for all four tumor types together. The mouse allograft model was not predictive. The human xenograft model was not predictive for breast or colon cancers, but was predictive for non-small cell lung and ovarian cancers when panels of xenografts were used. CONCLUSIONS: These results suggest that under the right framework and when panels are used, the in vitro cell line and human xenograft models may be useful in predicting the Phase II clinical trial performance of cancer drugs. Murine allograft models, as used in this analysis, appear of limited utility. PMID- 14519651 TI - Vaccination against B-cell chronic lymphocytic leukemia with trioma cells: preclinical evaluation. AB - PURPOSE: Trioma cells are lymphoma cells that have been fused to a hybridoma and have thereby been modified to express an immunoglobulin directed against surface receptors of antigen-presenting cells. Trioma cells that potentially include all lymphoma-derived antigens will be targeted to professional antigen-presenting cells in vivo. This allows uptake, processing, and presentation of tumor-derived antigens to T lymphocytes. In a mouse model, vaccination with trioma cells conferred long-lasting, T cell-dependent tumor immunity and was even able to eradicate established lymphomas. Here, we investigated whether this potent approach is effective in the human system. EXPERIMENTAL DESIGN: Malignant cells from 11 patients with B cell chronic-lymphocytic leukemia (B-CLL) were fused to an anti-Fc receptor hybridoma. The resulting trioma cells were extensively characterized with respect to their clonal origin. The induction of autologous tumor-specific T lymphocytes in the presence of trioma and antigen-presenting cells was examined in vitro by determining cytokine secretion in coculture assays. RESULTS: In seven cases, trioma cells could successfully be generated from B-CLL cells. Stimulation of autologous lymphocytes with trioma cells induced a leukemia-specific T-cell response. Immunostimulatory trioma cells were also obtained from two patients with solid B-cell lymphoma. CONCLUSIONS: Trioma mediated immunization may be a promising adjuvant treatment of human malignancies of the B-cell lineage, particularly of B-CLL, which has still a very poor prognosis. Our in vitro results pave the way for clinical application. PMID- 14519652 TI - Epidermal growth factor receptor VIII peptide vaccination is efficacious against established intracerebral tumors. AB - PURPOSE: The epidermal growth factor receptor (EGFR) is often amplified and structurally rearranged in malignant gliomas and other tumors such as breast and lung, with the most common mutation being EGFRvIII. In the study described here, we tested in mouse models a vaccine consisting of a peptide encompassing the tumor-specific mutated segment of EGFRvIII (PEP-3) conjugated to keyhole limpet hemocyanin [KLH (PEP-3-KLH)]. EXPERIMENTAL DESIGN: C57BL/6J or C3H mice were vaccinated with PEP-3-KLH and subsequently challenged either s.c. or intracerebrally with a syngeneic melanoma cell line stably transfected with a murine homologue of EGFRvIII. Control mice were vaccinated with KLH. To test its effect on established tumors, C3H mice were also challenged intracerebrally and subsequently vaccinated with PEP-3-KLH. RESULTS: S.c. tumors developed in all of the C57BL/6J mice vaccinated with KLH in Freund's adjuvant, and there were no long-term survivors. Palpable tumors never developed in 70% of the PEP-3-KLH vaccinated mice. In the C57BL/6J mice receiving the PEP-3-KLH vaccine, the tumors that did develop were significantly smaller than those in the control group (P < 0.05). PEP-3-KLH vaccination did not result in significant cytotoxic responses in standard cytotoxicity assays; however, antibody titers against PEP-3 were enhanced. The passive transfer of sera from the immunized mice to nonimmunized mice protected 31% of the mice from tumor development (P < 0.05). In vivo depletion studies showed that the effector cell population was natural killer and CD8+ T cells, and in vitro assays showed that macrophages could lyse target tumor cells with serum from the PEP-3-KLH-vaccinated mice. Peptide vaccination was also sufficiently potent to have marked efficacy against intracerebral tumors, resulting in a >173% increase in median survival time, with 80% of the C3H mice achieving long-term survival (P = 0.014). In addition, C3H mice with established intracerebral tumor that received a single treatment of PEP-3-KLH showed a 26% increase in median survival time, with 40% long-term survival (P = 0.007). CONCLUSIONS: Vaccination with an EGFRvIII-specific peptide is efficacious against both s.c. and established intracerebral tumors. The therapeutic effect of peptide vaccination may be mediated, in part, by antibody-dependent cellular cytotoxicity. PMID- 14519653 TI - Cisplatin down-regulation of cellular Fas-associated death domain-like interleukin-1beta-converting enzyme-like inhibitory proteins to restore tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis in human melanoma cells. AB - PURPOSE: Many melanoma cell lines and primary cultures are resistant to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis. In this study, we investigated the molecular mechanisms that control melanoma cell resistance and searched for chemotherapeutic drugs that could overcome the TRAIL resistance in melanoma cells. EXPERIMENTAL DESIGN: We examined 21 melanoma cell lines and 3 primary melanoma cultures for their sensitivity to TRAIL-induced apoptosis, and then tested cisplatin, chemptothecin, and etoposide for their synergistic effects on TRAIL sensitivity in resistant melanoma cells. RESULTS: Of 21 melanoma cell lines, 11 showed various degrees of sensitivity to TRAIL-induced apoptosis through caspase-8-initiated cleavage of caspase-3 and DNA fragmentation factor 45. The remaining cell lines and primary cultures were resistant to TRAIL, but cisplatin, chemptothecin, and etoposide sensitized the resistant cell lines and primary cultures to TRAIL-induced apoptosis, which also occurred through the caspase-8-initiated caspase cascade. Of the two TRAIL death receptors (DR4 and DR5), melanoma cells primarily expressed DR5 on cell surface. Cisplatin treatment had no effects on cell surface DR5 expression or intracellular expression of Fas associated death domain and caspase-8. Instead, cisplatin treatment down regulated intracellular expression of the short form of cellular Fas-associated death domain-like interleukin-1beta-converting enzyme-like inhibitory protein (c FLIP) and inhibited phosphorylation of the long form of c-FLIP. CONCLUSIONS: The results presented here indicate that cisplatin inhibits c-FLIP protein expression and phosphorylation to restore TRAIL-induced caspase-8-initiated apoptosis in melanoma cells, thus providing a new combined therapeutic strategy for melanomas. PMID- 14519654 TI - BCL-2 antisense oligonucleotide genasense is active against imatinib-resistant BCR-ABL-positive cells. AB - PURPOSE: The near-universal emergence of imatinib resistance in patients with acute forms of Philadelphia chromosome-positive leukemia highlights the need for additional therapy to control this disease. G3139 (Genasense, oblimersen; Genta Inc.), a Bcl-2 antisense oligonucleotide, has been shown to down-regulate the Bcl 2 protein and induce apoptosis in myeloid leukemia cells from treated patients. We tested G3139 for its ability to inhibit BCR-ABL-mediated transformation in mice. EXPERIMENTAL DESIGN: Nude mice (n = 5/group) were transplanted s.c. with imatinib-resistant BCR-ABL-transformed TF-1 cells (BCR-ABL-TF-1-R cells). Mice with established tumors (0.1 g) were treated for 14 days with G3139 (7 mg/kg/day i.p.), or with the reverse-sequence control oligonucleotide G3622 (7 mg/kg/day i.p.) or with imatinib (50 mg/kg/day i.p.). RESULTS: Mice treated with G3622 or imatinib died within 10-12 weeks. Nearly all of the mice treated with G3139 survived for >6 months and had reduced tumor volume. Three of the 5 mice showed complete tumor regression. A transient decrease in Bcl-2 protein was observed that correlated with histological evidence of apoptosis. In addition, we harvested BCR-ABL-TF-1-R tumor cells from mice treated with G3139 or control G3622 (7 mg/kg/day i.p., 7 days). Cells were then cultured with the antileukemic agents imatinib, daunorubicin, 1-beta-D-arabinofuranosylcytosine, or etoposide. G3139 pretreatment resulted in enhanced induction of apoptosis by all of the agents. CONCLUSION: These results suggest that G3139 is a promising candidate for treatment of patients with imatinib-resistant Ph-positive leukemia, and that combination of G3139 and imatinib may be useful to circumvent clinically acquired imatinib resistance. PMID- 14519655 TI - Reduction of stromal fibroblast-induced mammary tumor growth, by retroviral ribozyme transgenes to hepatocyte growth factor/scatter factor and its receptor, c-MET. AB - PURPOSE: Hepatocyte growth factor/scatter factor (HGF/SF) is known to increase the invasiveness and migration of cancer cells in vitro and induce angiogenesis. This study examined if inhibition of HGF/SF receptor expression by cancer cells and HGF/SF expression by stromal fibroblasts affects the growth of mammary cancer. EXPERIMENTAL DESIGN: Transgenes encoding ribozymes to specifically target human HGF/SF receptor (pLXSN-MET) or HGF/SF (pLXSN-HGF) were constructed using a pLXSN retroviral vector. Human mammary cancer cells MDA MB 231 was transduced with pLXSN-MET (MDA(+/+)). A human fibroblast cell line MRC5, which produces bioactive HGF/SF, was transduced with pLXSN-HGF (MRC5(+/+)). These cells were used in a nude mice breast tumor model. RESULTS: HGF receptor in MDA(+/+) cells and HGF in MRC5(+/+)cells were successfully removed with respective ribozymes as shown by reverse transcription-PCR and Western blotting, respectively. MDA(+/+) was found to have reduced invasiveness when stimulated with HGF/SF. MRC5(+/+) exhibited a significant reduction in HGF/SF production. When injected into athymic nude mice, MDA(+/+) exhibited a slower rate of growth, compared with the wild type (MDA(-/-)), and the cells transduced with control viral vector (MDA(+/ )). The growth of MDA(-/-) tumor was significantly enhanced when coimplanted with wild-type MRC5 (MRC5(-/-)), and the stimulatory effect was reduced when MRC5(+/+) cells were coimplanted instead of MRC5(-/-). The reduction of tumor growth was accompanied by reduction of angiogenesis, as demonstrated by the staining of VE cadherin in primary tumor tissues. CONCLUSIONS: Retroviral ribozyme transgenes targeting HGF/SF in fibroblasts or its receptor cMET in mammary cancer cells can reduce the growth of mammary cancer and associated angiogenesis by inhibiting paracrine stromal-tumor cell interactions. PMID- 14519656 TI - Identification of S100A2 as a target of the DeltaNp63 oncogenic pathway. AB - PURPOSE AND EXPERIMENTAL DESIGN: It has been proved recently that DeltaNp63 may play an oncogenic role in the tumorigenic pathway of squamous cell cancers. To gain additional insight into this pathway, we examined global patterns of gene expression in cancer cells after DeltaNp63 gene introduction using the oligonucleotide microarray approach. RESULTS: We found that S100A2 might be a target of the DeltaNp63 pathway. To confirm the data obtained from oligonucleotide microarray, we then examined the interaction of DeltaNp63 to S100A2. S100A2 induction was strictly dependent on DeltaNp63 expression by DeltaNp63 transgene and Northern analysis. DeltaNp63 transactivated the S100A2 promoter, and significantly more fold changes were seen in DeltaNp63-introduced cells than in p53-introduced cells, suggesting that DeltaNp63 may be a novel stimulator of the S100A2 promoter. CONCLUSION: Taken together, this evidence would seem to suggest that S100A2 is a novel downstream mediator of DeltaNp63. PMID- 14519657 TI - Speed kills: cellular and molecular bases of methamphetamine-induced nerve terminal degeneration and neuronal apoptosis. AB - Methamphetamine (METH) is a drug of abuse that has long been known to damage monoaminergic systems in the mammalian brain. Recent reports have provided conclusive evidence that METH can cause neuropathological changes in the rodent brain via apoptotic mechanisms akin to those reported in various models of neuronal death. The purpose of this review is to provide an interim account for a role of oxygen-based radicals and the participation of transcription factors and the involvement of cell death genes in METH-induced neurodegeneration. We discuss data suggesting the participation of endoplasmic reticulum and mitochondria mediated activation of caspase-dependent and -independent cascades in the manifestation of METH-induced apoptosis. Studies that use more comprehensive approaches to gene expression profiling should allow us to draw more instructive molecular portraits of the complex plastic and degenerative effects of this drug. PMID- 14519658 TI - Sphingosine-1-phosphate stimulates contraction of human airway smooth muscle cells. AB - The bioactive sphingolipid sphingosine-1-phosphate (S1P) that is increased in airways of asthmatic subjects markedly induced contraction of human airway smooth muscle (HASM) cells embedded in collagen matrices in a Gi-independent manner. Dihydro-S1P, which binds to S1P receptors, also stimulated contractility. S1P induced formation of stress fibers, contraction of individual HASM cells, and stimulated myosin light chain phosphorylation, which was inhibited by the Rho associated kinase inhibitor Y-27632. S1P-stimulated HASM cell contractility was independent of the ERK1/2 and PKC signaling pathways, important regulators of airway smooth muscle contraction. However, removal of extracellular calcium completely blocked S1P-mediated contraction and Y-27632 reduced it. S1P also induced calcium mobilization that was not desensitized by repeated additions. Pretreatment with thapsigargin to deplete InsP3-sensitive calcium stores partially blocked increases in [Ca2+]i induced by S1P, yet did not inhibit S1P stimulated contraction. In sharp contrast, the L-type calcium channel blocker verapamil markedly decreased S1P-induced HASM cell contraction, supporting a role for calcium influx from extracellular sources. Collectively, our results suggest that S1P may regulate HASM contractility, important in the pathobiology of asthma. PMID- 14519659 TI - Mechanical stretch activates nuclear factor-kappaB, activator protein-1, and mitogen-activated protein kinases in lung parenchyma: implications in asthma. AB - We investigated the effects of mechanical stretch and induced stimulation of lung parenchyma on the activation of proinflammatory transcription factors in normal mice and in a mouse model of asthma. Mechanical stretching of lung parenchyma led to increased activation of NF-kappaB and AP-1 transcription factors. Incubation of lung parenchyma with methacholine increased the activation of NF-kappaB, which was further augmented by stretch. Activation of NF-kappaB in response to mechanical stretch was associated with the phosphorylation and degradation of IkappaBalpha and the activation of IkappaB kinase. Stretch-induced activation of NF-kappaB involves activation of stretch-activated (SA) channels and the production of free radicals. Mechanical stretch and/or treatment with methacholine resulted in an increased activation of ERK1/2 and p38 MAP kinase, and the inhibition of the activity of these kinases partially blocked the stretch induced NF-kappaB and AP-1 activation. A greater level of NF-kappaB and ERK1/2 activity was observed in the asthmatic mice, which was further increased by mechanical stretching. The level of cyclooxygenase-2, an NF-kappaB-regulated enzyme, was also higher in lung parenchyma from asthmatic mice than in normal mice. Our data suggest that mechanical stretching of lung parenchyma activates NF kappaB and AP-1, at least in part, through the activation of MAP kinase signaling pathways. PMID- 14519660 TI - Serial analysis of gene expression in the skeletal muscle of endurance athletes compared to sedentary men. AB - Physical exercise produces several adaptive changes in skeletal muscle. However, the molecular mechanisms of these effects are poorly understood. We performed serial analysis of gene expression (SAGE) to quantify the global gene expression profile in sedentary and endurance-trained muscle. A total of 10869 SAGE tags was sequenced and represented 4727 genes. The genes most expressed in muscle are mainly involved in contraction and energy metabolism. Thirty-three genes were differentially expressed between endurance athletes and sedentary individuals. Four genes such as myosin binding protein C fast-type, glycogen phosphorylase, and pyruvate kinase were expressed less in endurance athletes, whereas eight genes coding for expressed sequence tag similar to (EST) crystallin alpha B, EST myosin light chain 2, EST surfactant pulmonary-associated protein A1, EST thrombospondin, EST fructose-bisphosphate aldolase A, EST cytochrome oxidase 1, NADH dehydrogenase 3, and G8 protein were up-regulated. Most of the up-regulated tags corresponded to novel genes. On the other hand, different isoforms of fructose-bisphosphate aldolase A were also differentially expressed. The current study underlying the most highly expressed genes allows a better understanding of global muscle characteristics in normal and endurance-trained individuals. Moreover, the current data suggest novel candidate genes that may be responsible for enhanced endurance performance. PMID- 14519661 TI - Electrophysiological and metabolic evidence that high-frequency stimulation of the subthalamic nucleus bridles neuronal activity in the subthalamic nucleus and the substantia nigra reticulata. AB - High-frequency stimulation (HFS) of the subthalamic nucleus (STN) has been shown to produce a dramatic alleviation of motor symptoms in patients with advanced Parkinson's disease. Its functional mechanism, however, remains obscure. We used extracellular recording and in situ cytochrome oxidase (CoI) mRNA hybridization to investigate the effects of HFS of the STN on neuronal activity of the STN and the substantia nigra reticulata (SNr) in normal rats and rats with 6 hydroxydopamine (6-OHDA) lesion of the substantia nigra compacta (SNc). To allow detection of spikes and analysis of firing activity, artifacts recorded during stimulation were scaled down using a template subtraction method. In both normal and lesioned rats, the activity of a majority of STN neurons was inhibited during stimulation. In the SNr, HFS also induced an inhibition of the activity of a majority of neurons in normal and lesioned rats. In situ hybridization histochemistry confirmed these results in that it showed a significant decrease in levels of CoI mRNA expression in the STN and SNr in both normal and lesioned rats during stimulation. These data afford an interesting insight into the functional mechanism of deep brain stimulation and support the hypothesis that HFS exerts an inhibitory influence on STN neuronal firing. PMID- 14519662 TI - PDE4D plays a critical role in the control of airway smooth muscle contraction. AB - The airways of mice deficient in the cAMP phosphodiesterase PDE4D gene are refractory to muscarinic cholinergic stimulation. This study was undertaken to determine whether altered smooth muscle contractility causes the PDE4D-/- phenotype. A major disruption in contractility was observed in isolated PDE4D-/- tracheas, with a 60% reduction in maximal tension and a fivefold decrease in sensitivity to muscarinic cholinergic agonists. Conversely, responses to KCl or arginine vasopressin were unaffected. PDE4D is the predominant PDE4 form in tracheal extracts and PDE4D mRNA is expressed in smooth muscle where muscarinic binding sites are most abundant. Cyclic AMP accumulation in response to acute G(s)alpha-coupled receptor stimulation was increased up to fourfold in the airway of PDE4D-/- mice when compared with wild-type. This increase in cAMP was associated with an increased sensitivity to PGE2-induced relaxation of the PDE4D /-tracheas. Furthermore, a blockade of prostanoid accumulation in PDE4D-/- tracheas restored the response to muscarinic cholinergic stimulation in vitro and in vivo. These results demonstrate that PDE4D plays a key role in balancing relaxant and contracting cues in airway smooth muscle, suggesting that natural mutations in the PDE4D gene have profound effects on airway tone. PMID- 14519663 TI - NFBD1/MDC1 regulates ionizing radiation-induced focus formation by DNA checkpoint signaling and repair factors. AB - NFBD1/MDC1 (mediator of DNA damage checkpoint 1) is a nuclear factor with an amino-terminal FHA (forkhead-associated) domain and a tandem repeat of BRCT (breast cancer susceptibility gene-1 carboxyl terminus) domains. We have previously shown that NFBD1 is an early participant in DNA damage signaling pathways and that ionizing radiation-induced nuclear foci (IRIF) of NFBD1 colocalize with several DNA checkpoint signaling and repair factors. We report here that NFBD1 physically associates with ATM, p53, components of the MRE11 RAD50-NBS1 (MRN) complex, and gamma-H2AX. An overexpressed FHA domain-containing fragment of NFBD1 binds to endogenous NFBD1 and components of the MRN complex, but not to gamma-H2AX. This fragment interferes with IRIF formation by endogenous NFBD1, MRE11, or NBS1. A BRCT domain-containing fragment of NFBD1 binds to gamma H2AX and 53BP1, but not to components of the MRN complex, and abolishes IRIF formation by NFBD1, MRE11, NBS1, 53BP1, CHK2 phospho-T68, gamma-H2AX, and possible ATM/ATR substrates recognized by anti-phospho-SQ/TQ antibody. These results suggest that NFBD1 is an ATM/ATR-dependent organizer that recruits DNA checkpoint signaling and repair proteins to the sites of DNA damage. PMID- 14519664 TI - Dexamethasone blocks the rapid biological effects of 17beta-estradiol in the rat uterus without antagonizing its global genomic actions. AB - Estrogens and glucocorticoids have opposing effects on the female reproductive tract, but the molecular basis for this antagonism is poorly understood. We therefore examined the biological and transcriptional programs induced by estrogens and glucocorticoids in the uterus of immature female rats. Estradiol 17beta (E2) rapidly induced morphological changes reminiscent of an acute inflammatory response, including infiltration of eosinophils, edema in the stroma and myometrium, and a decrease in the height of luminal epithelial cells, whereas dexamethasone (Dex) only altered stromal cell morphology. When coadministered with E2, Dex completely blocked the proinflammatory effects of E2. Surprisingly, examination of E2 and Dex effects on gene expression using cDNA microarrays and real-time PCR revealed that these hormones had similar effects on the expression of many genes and that very few genes displayed antagonistic regulation. Together, these results indicate strong discord between the early biologic and genomic actions of estrogens and glucocorticoids and highlight a complex regulatory role for glucocorticoids and GR in the mammalian uterus. PMID- 14519665 TI - Proinflammatory role of proteinase-activated receptor-2 in humans and mice during cutaneous inflammation in vivo. AB - Proteinase-activated receptor-2 belongs to a new subfamily of G-protein-coupled receptors. Its precise role during inflammation and the underlying mechanisms is still unclear. Our study establishes that PAR-2 plays a direct proinflammatory role during cutaneous inflammation in mice and humans in vivo. In a model of experimentally induced allergic (ACD) and toxic (ICD) contact dermatitis (CD) we show that ear swelling responses, plasma extravasation, and leucocyte adherence were significantly attenuated in PAR-2 null mutant (PAR-2-/-) mice compared with wild-type (PAR-2+/+) mice, especially at early stages. The proinflammatory effects by PAR-2 activation were significantly diminished using nitric oxide synthase inhibitors, while NF-kappaB and neuropeptides appear to play a minor role in these mechanisms. PAR-2-mediated up-regulation of E-selectin and cell adhesion molecule ICAM-1; enhanced plasma extravasation was observed in humans and mice and of interleukin-6 in mice in vivo. Thus, PAR-2 may be a beneficial therapeutic target for the treatment of inflammatory skin diseases. PMID- 14519666 TI - IL-1 induced release of Ca2+ from internal stores is dependent on cell-matrix interactions and regulates ERK activation. AB - The cellular mechanisms that modulate interleukin-1 (IL-1) signaling are not defined. In fibroblasts, IL-1 signaling is affected by the nature of cell-matrix adhesions including focal adhesions, adhesive domains that sequester IL-1 receptors. We conducted studies to elucidate which steps of cellular Ca2+ handling are affected by focal adhesions and by which mechanisms focal adhesions modulate IL-1-induced Ca2+ signals and ERK activation in human gingival fibroblasts. Cells were plated on poly-l-lysine or fibronectin and treated with tenascin, Hep-I, or SPARC peptides to inhibit focal adhesion formation. These treatments blocked IL-1 and thapsigargin-induced Ca2+ release from the endoplasmic reticulum, indicating that the ER-release pathway is focal adhesion dependent. Focal adhesions were also required for Ca2+ entry through store operated channels and for IL-1-induced ERK activation. Thus interactions with the extracellular matrix and focal adhesion formation regulate IL-1-induced generation of intracellular Ca2+ signals that in turn are required for ERK activation. PMID- 14519667 TI - Hypothalamic vasopressin release and hepatocyte Ca2+ signaling during liver regeneration: an interplay stimulating liver growth and bile flow. AB - Liver regeneration after partial hepatectomy is a plastic process during which the mechanisms that coordinate liver mass restoration compensate one another through a complex regulatory network of cytokines, growth factors, and hormones. Vasopressin, an agonist that triggers highly organized Ca2+ signals in the liver, may be one of these factors, although little in vivo evidence is available in support of this hypothesis. We provide evidence that hypothalamic vasopressin secretion is stimulated early after partial hepatectomy. Although hepatocytes were fully responsive to vasopressin during the first hours of regeneration, they became desensitized and exhibited slow oscillating Ca2+ responses to vasopressin on the following days. On the first day, hepatocyte V1a receptor density decreased and its lobular gradient increased in hepatectomized rats. By antagonizing the V1a receptor in vivo, we demonstrated that vasopressin contributes to NF-kappaB and cyclin (D1 and A) activation, to hepatocyte progression in the cell cycle, and to liver mass restoration. Finally, vasopressin exerted a choleretic effect shortly after hepatectomy, both in the isolated perfused liver and in the intact rat. In conclusion, we provide compelling in vivo evidence that vasopressin contributes significantly to growth initiation and bile flow stimulation in the early stages of liver regeneration. PMID- 14519668 TI - NOVH increases MMP3 expression and cell migration in glioblastoma cells via a PDGFR-alpha-dependent mechanism. AB - Nephroblastoma overexpressed gene (NOV) is highly expressed in the nervous system. We investigated its biological activity by expressing the human NOV gene (NOVH) in a human glioblastoma cell line that is negative for NOVH and by analyzing four clones with different levels of NOVH expression. There was no difference in cell proliferation between the NOVH-expressing cell lines, but there was increased cell adhesion and migration that correlated with increasing NOVH expression. Gene expression profiling was used to investigate the mechanisms by which NOVH expression regulated cell activity. We identified two induced genes in NOVH-expressing cells that are involved in cell migration: matrix metalloprotease (MMP)3 and platelet-derived growth factor receptor (PDGFR)-alpha. Our studies show that PDGFR-alpha induced MMP3 gene expression and increased cell proliferation and cell migration upon stimulation by platelet-derived growth factor (PDGF)-AA. We also show that the induction of MMP3 in cells expressing NOVH is potentiated by either cell density, serum, or PDGF-BB. Thus, expression of NOVH in glioblastoma cells triggers a cascade of gene expression resulting in increased cell adhesion and migration. PMID- 14519669 TI - VEGF-A and alphaVbeta3 integrin synergistically rescue angiogenesis via N-Ras and PI3-K signaling in human microvascular endothelial cells. AB - We recently showed that normal fibroblasts mediate capillary-like differentiation of human microvascular endothelial cells (HMVEC) in a 3-D angiogenesis model. Here, we show that a collaborative effect of VEGF-A and alphaVbeta3 integrin is critical in fibroblast-mediated angiogenesis because enhancement of both VEGF production by fibroblasts and beta3 integrin expression in HMVEC can rescue capillary-like endothelial differentiation under reduced serum conditions. To investigate the downstream signaling mechanisms, we compared N-Ras and Rho/Rac/Cdc42, as well as phosphatidylinositol 3-kinase (PI3-K) and Akt, for their involvement in the capillary-like network formation. The dominant-negative mutant of N-Ras (N-RasN17), but not the mutants of Rho/Rac/Cdc42, suppressed network formation. Overexpression of a constitutively active form of PI3-K rescued the network formation, which was inhibited by a dominant-negative >beta3 integrin; however, an active form of Akt failed to rescue the inhibition but induced a phenotypic change in HMVEC. Moreover, PI3-K is a downstream target of N Ras because it could be co-immunoprecipitated with N-Ras, and its active form could rescue the inhibitory effect of N-Ras N17. Thus, our data indicate the existence of N-Ras- and PI3-K-dependent but Rho/Rac/Cdc42- and Akt-independent signaling mechanisms for the synergistic effect of VEGF-A and alphaVbeta3 on fibroblast-mediated microvascular network formation. PMID- 14519670 TI - Alpha-synuclein implicated in Parkinson's disease is present in extracellular biological fluids, including human plasma. AB - Parkinson's disease (PD) and other related disorders are characterized by the accumulation of fibrillar aggregates of alpha-synuclein protein (alpha-syn) inside brain cells. It is likely that the formation of alpha-syn aggregates plays a seminal role in the pathogenesis of at least some of these diseases, because two different mutations in the gene encoding alpha-syn have been found in inherited forms of PD. alpha-Syn is mainly expressed by neuronal cells and is generally considered to exist as a cytoplasmic protein. Here, we report the unexpected identification of alpha-syn in conditioned culture media from untransfected and alpha-syn-transfected human neuroblastoma cells, as well as in human cerebrospinal fluid and blood plasma. The method used was immunocapture by using anti-alpha-syn antibodies coupled to magnetic beads, followed by detection on Western blots. In all cases, alpha-syn was identified as a single 15 kDa band, which co-migrated with a recombinant form of the protein and reacted with five different antibodies to alpha-syn. Our findings suggest that cells normally secrete alpha-syn into their surrounding media, both in vitro and in vivo. The detection of extracellular alpha-syn and/or its modified forms in body fluids, particularly in human plasma, offers new opportunities for the development of diagnostic tests for PD and related diseases. PMID- 14519671 TI - Similarities and differences in the responses of microorganisms to biocides. AB - Unlike antibiotics, biocides are multi-targeted antimicrobial agents. Several of the damaging effects reported to occur in the most widely studied organisms, bacteria, may also take place to varying degrees in other organisms. Nevertheless, there is considerable variation in the response of different microorganisms to biocides. Bacteria themselves (Gram-positive and Gram-negative vegetative organisms, mycobacteria and spores) respond differently to biocides and this disparity is widened when yeasts, moulds, protozoa and algae are considered. The underlying reasons for these varied responses are poorly understood at present, but the chemical composition of outer cellular layers is likely to be a factor of prime importance. Other possible contributory factors may be differences in stress responses, the presence of efflux pumps and cells occurring within biofilms or algal mats. PMID- 14519672 TI - Comparative activity of garenoxacin and other agents by susceptibility and time kill testing against Staphylococcus aureus, Streptococcus pyogenes and respiratory pathogens. AB - OBJECTIVES: Garenoxacin is a novel des-F(6)quinolone that has shown excellent antimicrobial activity against a wide range of clinically important microorganisms. In this study, its activity was examined, in comparison with that of other antimicrobial agents, by susceptibility and time-kill testing against Staphylococcus aureus, Streptococcus pyogenes and respiratory pathogens. METHODS: Overall, 200 bacterial strains were tested. The antimicrobial activity of garenoxacin was compared with that of ciprofloxacin, levofloxacin, moxifloxacin, amoxicillin, co-amoxiclav, cefuroxime, cefotaxime, ceftriaxone, imipenem, erythromycin and clarithromycin. In addition, the bactericidal activity of garenoxacin, moxifloxacin, levofloxacin and ciprofloxacin was evaluated by time kill analysis against four strains each of staphylococci [two methicillin susceptible (MSSA) and two methicillin-resistant (MRSA)], pneumococci (two penicillin-susceptible and two penicillin-resistant) and Streptococcus pyogenes (two erythromycin-susceptible and two erythromycin-resistant). Antibiotics were tested at concentrations 1-8 x MIC. RESULTS: MIC90 values of garenoxacin for the MSSA and MRSA strains were 0.03 and 2 mg/L, respectively. Among all the quinolones tested, garenoxacin yielded the lowest MIC values against all pneumococci (MIC90 0.12 mg/L) irrespective of macrolide resistance; the rank order of activity was garenoxacin> moxifloxacin>levofloxacin>ciprofloxacin. Excellent activity was shown also against Haemophilus influenzae (MIC90 or= 3 log10 decrease in viable counts (cfu/mL) within 3 h at 4 x MIC, whereas a moderate, slower killing rate was observed versus streptococci. CONCLUSIONS: This investigational des F(6)quinolone represents a promising alternative for the treatment of respiratory tract infections. PMID- 14519673 TI - Treatment failure in invasive aspergillosis: susceptibility of deep tissue isolates following treatment with amphotericin B. AB - OBJECTIVES: To determine whether treatment failure in invasive aspergillosis (IA) is the result of resistance of Aspergillus spp. isolates to amphotericin B. METHODS: Six Aspergillus fumigatus and six Aspergillus flavus isolates cultured from deep tissue biopsies in 11 patients with haematological malignancies during 1991-1998 were tested. A method based on the NCCLS M38-A broth microdilution method, with colorimetric determination of MICs, was used to determine the MICs of amphotericin B and itraconazole. RESULTS: All A. fumigatus isolates were susceptible to amphotericin B (MIC 0.25-0.5 mg/L), as were three A. flavus isolates (MIC 1 mg/L), but three were less susceptible (MIC 2 mg/L). All isolates were susceptible to itraconazole (MIC 0.125-0.25 mg/L). All patients had been treated with amphotericin B, having received a median of 12 days of treatment when the tissue was obtained. CONCLUSION: The difficulty in treating IA may not be because of the susceptibility of the isolates, but because of poor penetration of antifungal agents into infected tissue. Aspergillus spp. invade blood vessels causing thrombosis and tissue infarction, and therefore it may be difficult for antifungal drugs to exceed MICs in infected tissues. This highlights the need for different treatment strategies, such as surgery and the administration of cytokines. PMID- 14519674 TI - Telithromycin post-antibiotic and post-antibiotic sub-MIC effects for 10 Gram positive cocci. AB - Post-antibiotic effects (PAE) and post-antibiotic sub-MIC effects (PA-SME) of the ketolide telithromycin (HMR 3647) were determined for 10 Gram-positive cocci with various macrolide resistance mechanisms, including inducible and constitutive ribosomal methylase and macrolide efflux resistance genes. Strains tested included four Streptococcus pneumoniae, three Streptococcus pyogenes and three Staphylococcus aureus. Telithromycin MICs were 0.008-0.015 mg/L for pneumococci, 0.015-4.0 mg/L for S. pyogenes and 0.03 mg/L for staphylococci. PAE were determined after exposure of strains at 10 x MIC for 1 h. PA-SME were determined in the presence of 0.12x, 0.25x and 0.5x MIC of the agent after the initial 1 h exposure period. The PAE of telithromycin varied from 0.3 to 3.8 h; the PA-SME varied from 0.8 to 4.6 h, with maximal PA-SME varying from 1.5 to 4.6 h. PAE tended to be shortest for S. pyogenes (0.4-2.7 h) and S. aureus (0.3-2.4 h), compared to 1.5-3.8 h for S. pneumoniae. The duration of the PA-SME was similar for the three species tested. The low MICs for many strains as well as the PAE and PA-SME demonstrated in this study for telithromycin show promise for increasing the dosing interval of this ketolide, but will need verification by pharmacokinetic/pharmacodynamic and clinical studies. PMID- 14519675 TI - Adverse pregnancy outcome in users of sulfamethizole during pregnancy: a population-based observational study. AB - OBJECTIVE: To estimate the risk of adverse birth and neonatal outcome, and miscarriage in women who used sulfamethizole during pregnancy. METHODS: The association between use of sulfamethizole and adverse birth and neonatal outcome was investigated in a case-control and a cohort study in Denmark. We used data from the Prescription Database, the Birth Registry and the Hospital Discharge Registry in North Jutland County to study any association between sulfamethizole use and first recorded miscarriage. The cohort analysis included 3484 women who received a prescription for sulfamethizole from 30 days before conception to date of delivery, and 60175 women who did not use a sulphonamide-containing drug during pregnancy or 30 days before conception. The case-control analysis included 3347 women who had a miscarriage, of whom 90 had taken sulfamethizole, and 22599 primiparous controls who had a live birth. RESULTS: Among women who received prescriptions for sulfamethizole, adjusted odds ratios and 95% confidence intervals for adverse birth outcome were: malformation 1.17 (0.95-1.43); low birth weight 0.69 (0.49-0.98); pre-term birth 1.12 (0.97-1.30); stillbirth 1.02 (0.61-1.68); neonatal jaundice 1.14 (0.38-3.46); and for receiving a prescription for sulfamethizole within 1 week before miscarriage 1.66 (0.92-2.99). CONCLUSIONS: We found no increased risk of congenital malformation, stillbirth or pre-term birth, and no association between use of sulfamethizole late in pregnancy and risk of neonatal jaundice. There was an increased risk of miscarriage after exposure to sulfamethizole during the week before miscarriage, but further studies are needed to evaluate whether this increased risk is causal. PMID- 14519676 TI - Development of a microdilution method to evaluate Mycobacterium tuberculosis drug susceptibility. AB - A new rapid microdilution method, employing the dye resazurin as an indicator of mycobacterial growth, was developed to evaluate drug susceptibility of Mycobacterium tuberculosis reference strain H37Rv and of 13 M. tuberculosis susceptible or multidrug-resistant clinical strains. Different growth conditions were evaluated. The MICs of isoniazid, rifampicin, streptomycin and ethambutol were determined by the Microdilution Resazurin Assay (MRA) and the results compared with those obtained by the agar proportion method; complete agreement was always obtained. MRA resulted in a rapid, reliable, simple and inexpensive coloured method suitable for testing the susceptibility of M. tuberculosis clinical strains to first-line drugs; its employment in evaluating new antibacterial molecules is also suggested. PMID- 14519677 TI - Bacteriological outcome of combination versus single-agent treatment for staphylococcal endocarditis. AB - OBJECTIVE: To analyse the bacteriological outcome of combination versus single agent antimicrobial treatment in staphylococcal endocarditis. PATIENTS AND METHODS: Retrospective review of 152 episodes: 91 cases of native valve endocarditis (NVE), 74 due to Staphylococcus aureus and 17 due to coagulase negative staphylococci (CoNS); and 61 cases of prosthetic valve endocarditis (PVE), 29 due to S. aureus and 32 due to CoNS. RESULTS: Valves from patients with S. aureus NVE treated with any kind of combination antibiotic treatment were no more likely to be culture-negative than those treated with a single agent [19 (45%) of 42 versus 13 (41%) of 32; P = 0.69]. This finding remained unchanged when cases of CoNS NVE were added to the S. aureus group. In PVE, after adjusting for duration of treatment, valves from patients receiving any kind of combination treatment were 5.9 times (95% confidence interval 1.3-27.5) more likely to be culture-negative than those receiving monotherapy (P = 0.024). Patients treated for >14 days were more likely to be culture-negative than those treated for or=10 days) use of broad-spectrum combinations. Two factors were significantly associated with antibiotic prescribing practice, namely the country of origin (in those with >or=10 responders) and the level of microbiologist and/or infectious diseases specialist input. The greater the specialist input, the shorter the duration of therapy (P < 0.0001). CONCLUSIONS: The wide variability in antibiotic prescribing patterns suggests an urgent need to produce high-quality evidence to identify optimal antibiotic prescribing policies for bacteraemia in the critically ill patient. PMID- 14519682 TI - Gene expression variation in the adult human retina. AB - Despite evidence that differences in gene expression levels contribute significantly to phenotypic variation across individuals, there has been only limited effort to study gene expression variation in human tissue. To characterize expression variation in the normal human retina, we utilized a custom retinal microarray to analyze 33 normal retinas from 19 donors, aged 29-90 years. Statistical models were designed to separate and quantify biological and technical sources of variation, including age, gender, eye laterality, gene function and age-by-gender interaction. Although the majority of the 9406 genes analyzed showed relatively stable expression levels across different donors (for an average gene the expression level value of 95 out of a 100 individuals fell within a 1.23-fold range), 2.6% of genes showed significant donor-to-donor variation, with a false discovery rate of 10%. The mean expression ratio standard deviation was 0.15+/-0.8, log2, with a range of 0.09-0.99. Genes selectively expressed in photoreceptors showed higher expression variation than other gene classes. Gender, age and other donor-specific factors contributed significantly to the expression variation of multiple genes, and groups of genes with an age- and gender-associated expression pattern were identified. Our findings show that a significant fraction of gene expression variation in the normal human retina is attributable to identifiable biological factors. The greater expression variability of many genes central to retinal function (including photoreceptor specific genes) may be partially explained by the dynamics of the vision process, and raises the possibility that photoreceptor gene expression levels may contribute to phenotypic diversity across normal adult retinas. In addition, as such diversity may result in different levels of disease susceptibility, exploring its sources may provide insights into the pathogenesis of retinal disease. PMID- 14519683 TI - Expression profiling of FSHD muscle supports a defect in specific stages of myogenic differentiation. AB - The neuromuscular disorder facioscapulohumeral muscular dystrophy (FSHD) results from integral deletions of the subtelomeric repeat D4Z4 on chromosome 4q. A disruption of chromatin structure affecting gene expression is thought to underlie the pathophysiology. The global gene expression profiling of mature muscle tissue presented here provides the first insight into an FSHD-specific defect in myogenic differentiation. FSHD expression profiles generated by oligonucleotide microarrays were compared with those from normal muscle as well as other types of muscular dystrophies (DMD, aSGD) in order to determine FSHD specific changes. In addition, matched biopsies (affected and unaffected muscle) from individuals with FSHD served to monitor expression changes during the progression of the disease as well as to diminish non-specific changes resulting from individual variability. Among genes altered in an FSHD-specific and highly significant manner, many are involved in myogenic differentiation and suggest a partial block in the normal differentiation program. Indeed, many of the transcripts affected in FSHD represent direct targets of the transcription factor MyoD. Additional mis-expressed genes confirm a diminished capacity to buffer oxidative stress, as demonstrated in FSHD myoblasts. This enhanced vulnerability of proliferative stage myoblasts to reactive oxygen species is also disease specific, further implicating a defect in FSHD muscle satellite cells. Importantly, none of the genes localizing to the FSHD region at 4q35 were found to exhibit a significantly altered pattern of expression in FSHD muscle. This finding was corroborated by expression analysis of FSHD muscle using a custom cDNA microarray containing 51 genes and ESTs from the 4q35 region. Disruptions in FSHD myogenesis and oxidative capacity may therefore not arise from a position effect mechanism as has been previously suggested, but rather from a global effect on gene regulation. Improper nuclear localization of 4qter is discussed as an alternative model for FSHD gene regulation and pathogenesis. PMID- 14519684 TI - RING finger 1 mutations in Parkin produce altered localization of the protein. AB - The Parkin gene (PRKN) encodes an E3 protein-ubiquitin ligase for which loss of function is associated with autosomal-recessive juvenile (<20 years) and early onset Parkinsonism (<45 years). Although detailed pathological reports are scarce, brains from patients with homozygous exonic deletions demonstrate neuronal loss in the substantia nigra, albeit without the Lewy body pathology characteristic of idiopathic Parkinson's disease. However, there are rare descriptions of more florid pathology, including Lewy bodies and tau positive astrocytes in individuals with compound heterozygous mutations. In the present study we examined whether PRKN point mutations, leading to amino acid substitutions, may alter the cellular distribution of the protein produced. Wild type Parkin was homogeneously distributed throughout the cytoplasm with a small amount of protein in the nucleus after transfection into human embryonic kidney cells. Mutant isoforms with A82E, G328E and C431F amino acid substitutions were also normally distributed. However, two mutant isoforms, R256C and R275W, within RING finger 1 of the Parkin protein (238-293 amino acids), produced an unusual distribution of the protein, with large cytoplasmic and nuclear inclusions. We have replicated this observation in primary cultured neurons and demonstrate, by the accumulation/co-localization of cytoskeletal protein vimentin, that the inclusion bodies are aggresomes, a cellular response to misfolded protein. PMID- 14519685 TI - Polymorphism, shared functions and convergent evolution of genes with sequences coding for polyalanine domains. AB - Mutations causing expansions of polyalanine domains are responsible for nine hereditary diseases. Other GC-rich sequences coding for some polyalanine domains were found to be polymorphic in human. These observations prompted us to identify all sequences in the human genome coding for polyalanine stretches longer than four alanines and establish their degree of polymorphism. We identified 494 annotated human proteins containing 604 polyalanine domains. Thirty-two percent (31/98) of tested sequences coding for more than seven alanines were polymorphic. The length of the polyalanine-coding sequence and its GCG or GCC repeat content are the major predictors of polymorphism. GCG codons are over-represented in human polyalanine coding sequences. Our data suggest that GCG and GCC codons play a key role in polyalanine-coding sequence appearance and polymorphism. The grouping by shared function of polyalanine-containing proteins in Homo sapiens, Drosophila melanogaster and Caenorhabditis elegans shows that the majority are involved in transcriptional regulation. Phylogenetic analyses of HOX, GATA and EVX protein families demonstrate that polyalanine domains arose independently in different members of these families, suggesting that convergent molecular evolution may have played a role. Finally polyalanine domains in vertebrates are conserved between mammals and are rarer and shorter in Gallus gallus and Danio rerio. Together our results show that the polymorphic nature of sequences coding for polyalanine domains makes them prime candidates for mutations in hereditary diseases and suggests that they have appeared in many different protein families through convergent evolution. PMID- 14519686 TI - Analysis of mammalian proteins involved in chromatin modification reveals new metaphase centromeric proteins and distinct chromosomal distribution patterns. AB - We have examined the metaphase chromosomal localization of 15 proteins that have previously been described as involved in mammalian chromatin modification and/or transcriptional modulation. Immunofluorescence data indicate that all the proteins localize to human and mouse centromeres, a neocentromere, and the active centromere of a dicentric chromosome, with six of these proteins (Sin3A, PCAF, MYST, MBD2, ORC2, P300/CBP) being demonstrated at mammalian centromeres for the first time. Most of these proteins fall into two distinct chromosomal distribution patterns: (a) kinetochore-associated proteins (Sin3A, PCAF, MYST and BAF180), which colocalize with metaphase kinetochores, but not any of the pericentric and other major heterochromatic regions; and (b) heterochromatin associated proteins (MeCP2, MBD1, MBD2, ATRX, HP1alpha, HDAC1, HDAC2, DNMT1 and DNMT3b), which colocalize with centromeric/pericentric heterochromatin and all other major heterochromatic sites. A heterogeneous third group (c) consists of the origin recognition complex subunit ORC2 and the histone acetyltransferase P300/CBP, which associate generally with kinetochores in humans and centromeric/pericentric heterochromatin in mouse, with some minor differences in localization. These observations indicate an extensive sharing of protein components involved in chromatin modification at gene loci, centromeres and various chromosomal heterochromatic landmarks. The definition of distinct patterns of chromosomal distribution for these proteins provides a useful basis for the further investigation of the broad-ranging roles of these proteins. PMID- 14519687 TI - The (CGG)n repeat element within the 5' untranslated region of the FMR1 message provides both positive and negative cis effects on in vivo translation of a downstream reporter. AB - The human fragile X mental retardation 1 (FMR1) gene contains a polymorphic (CGG) trinucleotide repeat element in its 5' untranslated region. Expansion of the (CGG)n element beyond 200 repeats (full mutation range) generally leads to transcriptional silencing; consequent loss of the FMR1 protein (FMRP) results in fragile X syndrome, the most frequent form of inherited mental impairment. For carriers of smaller expansions (55< or =n< or =200; premutation range), FMRP levels are gradually reduced with increasing repeat number, despite elevated FMR1 mRNA levels, suggesting that translation is impeded within the premutation range. To examine in more detail the influence of the CGG repeat on translation, CMV immediate-early promoter constructs, containing the FMR1 5'-UTR with various (CGG)n repeat lengths (0< or =n< or =99) and a downstream (luciferase) reporter, were transfected into two human cell lines, a neural cell-derived line (SK) and a fetal kidney cell-derived line (293). For both cell types, the CGG element exerts distinct effects on reporter expression, depending on the length of the repeat. For n> or =30, luciferase expression decreases with increasing repeat length, consistent with earlier observations of decreased FMRP expression in peripheral blood leucocytes over the same repeat range, despite a slight increase in mRNA level for the larger repeats. Surprisingly, for smaller alleles (0< or =n< or =30), reporter expression actually increases by nearly two-fold with increasing repeat length in the absence of any change in mRNA level. These results suggest that the CGG repeat element can exert both positive (n<30) and negative (n>30) effects on translation. Interestingly, optimal translation appears to occur near the modal repeat number within the general human population. PMID- 14519689 TI - Multiple quantitative trait loci modify the heart failure phenotype in murine cardiomyopathy. AB - The variability in outcome of heart failure patients depends on a number of factors including differences in their genetic background. To identify novel genes that modify the human heart failure phenotype, we used a strategy of quantitative trait locus (QTL) mapping in an experimental mouse model of dilated cardiomyopathy induced by cardiac-specific overexpression of calsequestrin and characterized by a strong strain-specific variability in the phenotype. We identified two novel QTLs, Hrtfm3 (heart failure modifier 3) on chromosome (Chr) 4 and Hrtfm4 on Chr 18, significantly linked to survival with likelihood ratio statistics (LRS) of 19.9 and 23.6 respectively (corresponding to LOD scores of 4.3 and 5.1). Two other QTLs, Hrtfm5 on Chr 2 and Hrtfm6 on Chr 13, were significantly linked to cardiac function as measured by echocardiographic fractional shortening (LRS 22.1 and 15.2 respectively, LOD score 4.8 and 3.3) and left ventricular end-diastolic diameter (LRS 23.5 and 18.8, LOD score 5.1 and 4.1). Importantly, Hrtfm5 was not significantly linked to survival. A significant interaction was found between Hrtfm4 and two other QTLs (Hrtfm6 and a QTL near to the marker D19Mit88) for fractional shortening with a LRS of 34.6 and 26.5 respectively (LOD score 7.5 and 5.8). These data show that the effect of genetic background on murine heart failure is complex and result from the action of several loci that differentially modify the cardiac phenotype. The identification of these novel modifier genes will serve as strong candidates for the discovery of modifiers in human heart failure. PMID- 14519690 TI - A piece of my mind. Reflections while listening to the Glazunov Saxophone Concerto. PMID- 14519688 TI - Mouse models of USH1C and DFNB18: phenotypic and molecular analyses of two new spontaneous mutations of the Ush1c gene. AB - We mapped two new recessive mutations causing circling behavior and deafness to the same region on chromosome 7 and showed they are allelic by complementation analysis. One was named 'deaf circler' (allele symbol dfcr) and the other 'deaf circler 2 Jackson' (allele symbol dfcr-2J). Both were shown to be mutations of the Ush1c gene, the mouse ortholog of the gene responsible for human Usher syndrome type IC and for the non-syndromic deafness disorder DFNB18. The Ush1c gene contains 28 exons, 20 that are constitutive and eight that are alternatively spliced. The dfcr mutation is a 12.8 kb intragenic deletion that eliminates three constitutive and five alternatively spliced exons. The dfcr-2J mutation is a 1 bp deletion in an alternatively spliced exon that creates a transcriptional frame shift, changing 38 amino acid codons before introducing a premature stop codon. Both mutations cause congenital deafness and severe balance deficits due to inner ear dysfunction. The stereocilia of cochlear hair cells are disorganized and splayed in mutant mice, with subsequent degeneration of the hair cells and spiral ganglion cells. Harmonin, the protein encoded by Ush1c, has been shown to bind, by means of its PDZ-domains, with the products of other Usher syndrome genes, including Myo7a, Cdh23 and Sans. The complexes formed by these protein interactions are thought to be essential for maintaining the integrity of hair cell stereocilia. The Ush1c mutant mice described here provide a means to directly investigate these interactions in vivo and to evaluate gene structure function relationships that affect inner ear and eye phenotypes. PMID- 14519691 TI - Old drugs for a new bug: influenza, HIV drugs enlisted to fight SARS. PMID- 14519692 TI - SARS thrusts quarantine into the limelight. PMID- 14519693 TI - SARS attack: reality vs perception collide during students' Beijing visit. PMID- 14519694 TI - Virtual patients help medical students link basic science with clinical care. PMID- 14519697 TI - Reliability and validity of prostate-specific antigen. PMID- 14519698 TI - Effects of estrogen plus progestin on risk of dementia. PMID- 14519699 TI - Effects of estrogen plus progestin on risk of dementia. PMID- 14519700 TI - Effects of estrogen plus progestin on risk of dementia. PMID- 14519701 TI - Effects of estrogen plus progestin on risk of dementia. PMID- 14519702 TI - Passive smoke exposure and risk of death from coronary heart disease. PMID- 14519703 TI - Adipocytokines in synovial fluid. PMID- 14519704 TI - Natural vs vaccine-acquired immunity to cytomegalovirus. PMID- 14519705 TI - Timing of initial cereal exposure in infancy and risk of islet autoimmunity. AB - CONTEXT: Dietary exposures in infancy have been implicated, albeit inconsistently, in the etiology of type 1 diabetes mellitus (DM). OBJECTIVE: To examine the association between cereal exposures in the infant diet and appearance of islet autoimmunity (IA). DESIGN: Birth cohort study conducted from 1994 to 2002 with a mean follow-up of 4 years. SETTING: Newborn screening for HLA was done at St Joseph's Hospital in Denver, Colo. First-degree relatives of type 1 DM individuals were recruited from the Denver metropolitan area. PARTICIPANTS: We enrolled 1183 children at increased type 1 DM risk, defined as either HLA genotype or having a first-degree relative with type 1 DM, at birth and followed them prospectively. We obtained exposure and outcome measures for 76% of enrolled children. Participants had variable lengths of follow-up (9 months to 9 years). MAIN OUTCOME MEASURES: Blood draws for the detection of insulin autoantibody, glutamic acid decarboxylase autoantibody, or IA-2 autoantibody were performed at 9, 15, and 24 months and annually thereafter. Children with IA (n = 34) were defined as those testing positive for at least 1 of the autoantibodies on 2 or more consecutive visits and who tested positive or had diabetes on their most recent visit. RESULTS: Children initially exposed to cereals between ages 0 and 3 months (hazard ratio [HR], 4.32; 95% confidence interval [CI], 2.0-9.35) and those who were exposed at 7 months or older (HR, 5.36; 95% CI, 2.08-13.8) had increased hazard of IA compared with those who were exposed during the fourth through sixth month, after adjustment for HLA genotype, family history of type 1 DM, ethnicity, and maternal age. In children who were positive for the HLA DRB1*03/04,DQB8 genotype, adjusted HRs were 5.55 (95% CI, 1.92-16.03) and 12.53 (95% CI, 3.19-49.23) for initial cereal exposure between ages 0 to 3 months and at 7 months or older, respectively. CONCLUSION: There may be a window of exposure to cereals in infancy outside which initial exposure increases IA risk in susceptible children. PMID- 14519706 TI - Early infant feeding and risk of developing type 1 diabetes-associated autoantibodies. AB - CONTEXT: Dietary factors modifying type 1 diabetes mellitus (DM) risk have been proposed, but little is known if they trigger the islet autoimmunity that precedes clinical disease. OBJECTIVE: To determine whether breastfeeding duration, food supplementation, or age at introduction of gluten-containing foods influences the risk of developing islet autoantibodies. DESIGN AND SETTING: Prospective natural history cohort study conducted from 1989 to 2003 in inpatient/outpatient clinics in Germany. PARTICIPANTS: The BABYDIAB study follows newborn children of parents with type 1 DM. Eligibility requirements were met in 1610 children. Blood samples were obtained at birth, age 9 months, 2, 5, and 8 years. Dropout rate was 14.4% by age 5 years. Breastfeeding data were obtained by prospective questionnaires (91% complete), and food supplementation data were obtained by family interview (72% for food supplementation and 80% for age of gluten introduction). MAIN OUTCOME MEASURE: Development of islet autoantibodies (insulin, glutamic acid decarboxylase, or IA-2 antibodies) in 2 consecutive blood samples. RESULTS: Life-table islet autoantibody frequency was 5.8% (SE, 0.6%) by age 5 years. Reduced total or exclusive breastfeeding duration did not significantly increase the risk of developing islet autoantibodies. Food supplementation with gluten-containing foods before age 3 months, however, was associated with significantly increased islet autoantibody risk (adjusted hazard ratio, 4.0; 95% confidence interval, 1.4-11.5; P =.01 vs children who received only breast milk until age 3 months). Four of 17 children who received gluten foods before age 3 months developed islet autoantibodies (life-table 5-year risk, 24%; SE, 10%). All 4 children had the high-risk DRB1*03/04,DQB1*0302 genotype. Early exposure to gluten did not significantly increase the risk of developing celiac disease-associated autoantibodies. Children who first received gluten foods after age 6 months did not have increased risks for islet or celiac disease autoantibodies. CONCLUSION: Ensuring compliance to infant feeding guidelines is a possible way to reduce the risk of development of type 1 DM autoantibodies. PMID- 14519707 TI - Effects of estrogen plus progestin on risk of fracture and bone mineral density: the Women's Health Initiative randomized trial. AB - CONTEXT: In the Women's Health Initiative trial of estrogen-plus-progestin therapy, women assigned to active treatment had fewer fractures. OBJECTIVE: To test the hypothesis that the relative risk reduction of estrogen plus progestin on fractures differs according to risk factors for fractures. DESIGN, SETTING, AND PARTICIPANTS: Randomized controlled trial (September 1993-July 2002) in which 16 608 postmenopausal women aged 50 to 79 years with an intact uterus at baseline were recruited at 40 US clinical centers and followed up for an average of 5.6 years. INTERVENTION: Women were randomly assigned to receive conjugated equine estrogen, 0.625 mg/d, plus medroxyprogesterone acetate, 2.5 mg/d, in 1 tablet (n = 8506) or placebo (n = 8102). MAIN OUTCOME MEASURES: All confirmed osteoporotic fracture events that occurred from enrollment to discontinuation of the trial (July 7, 2002); bone mineral density (BMD), measured in a subset of women (n = 1024) at baseline and years 1 and 3; and a global index, developed to summarize the balance of risks and benefits to test whether the risk-benefit profile differed across tertiles of fracture risk. RESULTS: Seven hundred thirty-three women (8.6%) in the estrogen-plus-progestin group and 896 women (11.1%) in the placebo group experienced a fracture (hazard ratio [HR], 0.76; 95% confidence interval [CI], 0.69-0.83). The effect did not differ in women stratified by age, body mass index, smoking status, history of falls, personal and family history of fracture, total calcium intake, past use of hormone therapy, BMD, or summary fracture risk score. Total hip BMD increased 3.7% after 3 years of treatment with estrogen plus progestin compared with 0.14% in the placebo group (P<.001). The HR for the global index was similar across tertiles of the fracture risk scale (lowest fracture risk tertile, HR, 1.20; 95% CI, 0.93-1.58; middle tertile, HR, 1.23; 95% CI, 1.04-1.46; highest tertile, HR, 1.03; 95% CI, 0.88-1.24) (P for interaction =.54). CONCLUSIONS: This study demonstrates that estrogen plus progestin increases BMD and reduces the risk of fracture in healthy postmenopausal women. The decreased risk of fracture attributed to estrogen plus progestin appeared to be present in all subgroups of women examined. When considering the effects of hormone therapy on other important disease outcomes in a global model, there was no net benefit, even in women considered to be at high risk of fracture. PMID- 14519708 TI - Effects of estrogen plus progestin on gynecologic cancers and associated diagnostic procedures: the Women's Health Initiative randomized trial. AB - CONTEXT: The effects of continuous combined hormone therapy on gynecologic cancers have not been investigated previously in a randomized trial setting. OBJECTIVE: To determine the possible associations of estrogen plus progestin on gynecologic cancers and related diagnostic procedures. DESIGN, SETTING, AND PARTICIPANTS: Randomized, double-blind, placebo-controlled trial of 16 608 postmenopausal women, who had not had a hysterectomy at baseline and who had been recruited from 40 US clinical centers between September 1993 and October 1998 (average follow-up, 5.6 years). INTERVENTION: One tablet per day containing 0.625 mg of conjugated equine estrogens plus 2.5 mg of medroxyprogesterone acetate (n = 8506) or placebo (n = 8102). MAIN OUTCOME MEASURE: Incident invasive cancer of the ovary and endometrium. RESULTS: In 5.6 years of follow-up, there were 32 cases of invasive ovarian cancer, 58 cases of endometrial cancer, 1 case of nonendometrial uterine cancer, 13 cases of cervical cancer, and 7 cases of other gynecologic cancers. The hazard ratio (HR) for invasive ovarian cancer in women assigned to estrogen plus progestin compared with placebo was 1.58 (95% confidence interval [CI], 0.77-3.24). The HR for endometrial cancer was 0.81 (95% CI, 0.48-1.36). No appreciable differences were found in the distributions of tumor histology, stage, or grade for either cancer site. The incidence of other gynecologic cancers was low and did not differ by randomization assignment. More women taking estrogen plus progestin required endometrial biopsies (33% vs 6%; P<.001). CONCLUSIONS: This randomized trial suggests that continuous combined estrogen plus progestin therapy may increase the risk of ovarian cancer while producing endometrial cancer rates similar to placebo. The increased burden of endometrial biopsies required to assess vaginal bleeding further limits the acceptability of this regimen. These data provide additional support for caution in the use of continuous combined hormones. PMID- 14519709 TI - Azithromycin in patients with cystic fibrosis chronically infected with Pseudomonas aeruginosa: a randomized controlled trial. AB - CONTEXT: Treatment strategies for cystic fibrosis (CF) lung disease include antibiotics, mucolytics, and anti-inflammatory therapies. Increasing evidence suggests that macrolide antibiotics might be beneficial in patients with CF. OBJECTIVE: To determine if an association between azithromycin use and pulmonary function exists in patients with CF. DESIGN AND SETTING: A multicenter, randomized, double-blind, placebo-controlled trial conducted from December 15, 2000, to May 2, 2002, at 23 CF care centers in the United States. PARTICIPANTS: Of the 251 screened participants with a diagnosis of CF, 185 (74%) were randomized. Eligibility criteria included age 6 years or older, infection with Pseudomonas aeruginosa for 1 or more years, and a forced expiratory volume in 1 second (FEV1) of 30% or more. Participants were stratified by FEV1 (> or =60% predicted vs <60% predicted), weight of less than 40 kg vs 40 kg or more, and CF center. INTERVENTION: The active group (n = 87) received 250 mg (weight <40 kg) or 500 mg (weight > or =40 kg) of oral azithromycin 3 days a week for 168 days; placebo group (n = 98) received identically packaged tablets. MAIN OUTCOME MEASURES: Change in FEV1 from day 0 to completion of therapy at day 168 and determination of safety. Secondary outcomes included pulmonary exacerbations and weight gain. RESULTS: The azithromycin group had a mean 0.097-L (SD, 0.26) increase in FEV1 at day 168 compared with 0.003 L (SD, 0.23) in the placebo group (mean difference, 0.094 L; 95% confidence interval [CI], 0.023-0.165; P =.009). Nausea occurred in 17% more participants in the azithromycin group (P =.01), diarrhea in 15% more (P =.009), and wheezing in 13% more (P =.007). Participants in the azithromycin group had less risk of experiencing an exacerbation than participants in the placebo group (hazard ratio, 0.65; 95% CI, 0.44-0.95; P =.03) and weighed at the end of the study an average 0.7 kg more than participants receiving placebo (95% CI, 0.1-1.4 kg; P =.02). CONCLUSION: Azithromycin treatment was associated with improvement in clinically relevant end points and should be considered for patients with CF who are 6 years or older and chronically infected with P aeruginosa. PMID- 14519710 TI - Analgesic effect of the synthetic cannabinoid CT-3 on chronic neuropathic pain: a randomized controlled trial. AB - CONTEXT: 1',1'dimethylheptyl-Delta8-tetrahydrocannabinol-11-oic acid (CT-3), a potent analog of THC-11-oic acid, produces marked antiallodynic and analgesic effects in animals without evoking the typical effects described in models of cannabinoids. Therefore, CT-3 may be an effective analgesic for poorly controlled resistant neuropathic pain. OBJECTIVE: To examine the analgesic efficacy and safety of CT-3 in chronic neuropathic pain in humans. DESIGN AND SETTING: Randomized, placebo-controlled, double-blind crossover trial conducted in Germany from May-September 2002. PARTICIPANTS: Twenty-one patients (8 women and 13 men) aged 29 to 65 years (mean, 51 years) who had a clinical presentation and examination consistent with chronic neuropathic pain (for at least 6 months) with hyperalgesia (n = 21) and allodynia (n = 7). INTERVENTIONS: Patients were randomized to two 7-day treatment orders in a crossover design. Two daily doses of CT-3 (four 10-mg capsules per day) or identical placebo capsules were given during the first 4 days and 8 capsules per day were given in 2 daily doses in the following 3 days. After a washout and baseline period of 1 week each, patients crossed over to the second 7-day treatment period. MAIN OUTCOME MEASURES: Visual analog scale (VAS) and verbal rating scale scores for pain; vital sign, hematologic and blood chemistry, and electrocardiogram measurements; scores on the Trail-Making Test and the Addiction Research Center Inventory-Marijuana scale; and adverse effects. RESULTS: The mean differences over time for the VAS values in the CT-3-placebo sequence measured 3 hours after intake of study drug differed significantly from those in the placebo-CT-3 sequence (mean [SD], -11.54 [14.16] vs 9.86 [21.43]; P =.02). Eight hours after intake of the drug, the pain scale differences between groups were less marked. No dose response was observed. Adverse effects, mainly transient dry mouth and tiredness, were reported significantly more often during CT-3 treatment (mean [SD] difference, -0.67 [0.50] for CT-3-placebo sequence vs 0.10 [0.74] for placebo-CT-3 sequence; P =.02). There were no significant differences with respect to vital signs, blood tests, electrocardiogram, Trail-Making Test, and Addiction Research Center Inventory-Marijuana scale. No carryover or period effects were observed except on the Trail-Making Test. CONCLUSIONS: In this preliminary study, CT-3 was effective in reducing chronic neuropathic pain compared with placebo. No major adverse effects were observed. PMID- 14519711 TI - Association between thimerosal-containing vaccine and autism. AB - CONTEXT: Mercuric compounds are nephrotoxic and neurotoxic at high doses. Thimerosal, a preservative used widely in vaccine formulations, contains ethylmercury. Thus it has been suggested that childhood vaccination with thimerosal-containing vaccine could be causally related to neurodevelopmental disorders such as autism. OBJECTIVE: To determine whether vaccination with a thimerosal-containing vaccine is associated with development of autism. DESIGN, SETTING, AND PARTICIPANTS: Population-based cohort study of all children born in Denmark from January 1, 1990, until December 31, 1996 (N = 467 450) comparing children vaccinated with a thimerosal-containing vaccine with children vaccinated with a thimerosal-free formulation of the same vaccine. MAIN OUTCOME MEASURES: Rate ratio (RR) for autism and other autistic-spectrum disorders, including trend with dose of ethylmercury. RESULTS: During 2 986 654 person-years, we identified 440 autism cases and 787 cases of other autistic-spectrum disorders. The risk of autism and other autistic-spectrum disorders did not differ significantly between children vaccinated with thimerosal-containing vaccine and children vaccinated with thimerosal-free vaccine (RR, 0.85 [95% confidence interval [CI], 0.60-1.20] for autism; RR, 1.12 [95% CI, 0.88-1.43] for other autistic-spectrum disorders). Furthermore, we found no evidence of a dose-response association (increase in RR per 25 microg of ethylmercury, 0.98 [95% CI, 0.90-1.06] for autism and 1.03 [95% CI, 0.98-1.09] for other autistic-spectrum disorders). CONCLUSION: The results do not support a causal relationship between childhood vaccination with thimerosal containing vaccines and development of autistic-spectrum disorders. PMID- 14519712 TI - Diagnosis and management of female infertility. PMID- 14519713 TI - Infant diets and type 1 diabetes: too early, too late, or just too complicated? PMID- 14519714 TI - MSJAMA. Roles for medical students in the political process. PMID- 14519715 TI - MSJAMA. State ballot initiatives for health care reform. PMID- 14519716 TI - MSJAMA. Policies to regulate gifts to physicians from industry. PMID- 14519717 TI - MSJAMA. Educational and legislative initiatives affecting residency training in abortion. PMID- 14519718 TI - JAMA patient page. Infant feeding. PMID- 14519719 TI - Analysis of automatically generated peptide mass fingerprints of cellular proteins and antigens from Helicobacter pylori 26695 separated by two-dimensional electrophoresis. AB - Helicobacter pylori is a causative agent of severe diseases of the gastric tract ranging from chronic gastritis to gastric cancer. Cellular proteins of H. pylori were separated by high resolution two-dimensional gel electrophoresis. A dataset of 384 spots was automatically picked, digested, spotted, and analyzed by matrix assisted laser desorption ionization mass spectrometry peptide mass fingerprint in triple replicates. This procedure resulted in 960 evaluable mass spectra. Using a new version of our data analysis software MS-Screener we improved identification and tested reliability of automatically generated data by comparing with manually produced data. Antigenic proteins from H. pylori are candidates for vaccines and diagnostic tests. Previous immunoproteomics studies of our group revealed antigen candidates, and 24 of them were now closely analyzed using the MS-Screener software. Only in three spots minor components were found that may have influenced their antigenicities. These findings affirm the value of immunoproteomics as a hypothesis-free approach. Additionally, the protein species distribution of the known antigen GroEL was investigated, dimers of the protein alkyl hydroperoxide reductase were found, and the fragmentation of gamma-glutamyltranspeptidase was demonstrated. PMID- 14519720 TI - Functional interaction trap: a strategy for validating the functional consequences of tyrosine phosphorylation of specific substrates in vivo. AB - Protein tyrosine phosphorylation controls diverse signaling pathways, and disregulated tyrosine kinase activity plays a direct role in human diseases such as cancer. Because activated kinases exert their effects by phosphorylating multiple substrate proteins, it is difficult or impossible to assess experimentally the contribution of a particular substrate to a cellular response or activity. To overcome this problem, we have developed a novel approach termed the "functional interaction trap," in which two proteins are induced to interact in a pairwise fashion through an engineered, highly specific binding interface. We show that the functional interaction trap can be used to direct a modified tyrosine kinase to specifically phosphorylate a single substrate of choice in vivo, permitting analysis of the resulting biological output. This strategy provides a powerful tool for validating the functional significance of tyrosine phosphorylation and other post-translational modifications identified by proteomic discovery efforts. PMID- 14519721 TI - How to reduce the toll of road traffic accidents. PMID- 14519722 TI - Atypical manifestations of gastro-oesophageal reflux. PMID- 14519723 TI - Interferon alpha treatment and endocrine disease. PMID- 14519724 TI - Clocks, genes and sleep. PMID- 14519725 TI - The National Health Service and the internet. PMID- 14519726 TI - Enteral stenting in 21 patients with malignant gastroduodenal obstruction. AB - Placement of an enteral stent is emerging as an effective alternative to surgery for symptomatic relief in patients with malignant gastric outflow and duodenal obstruction. We report experience in a series of patients who had the Wallstent enteral prosthesis inserted. 21 consecutive patients with inoperable obstruction, median age 75 years, had twenty-three procedures for insertion of the Wallstent under fluoroscopic guidance. Stent implantation was successful in 18 patients (86%), all of whom gained relief from nausea and vomiting. There were no short term complications. Median survival after the procedure was four months (longest ten months), and no deaths were related to insertion. The efficacy and lack of complications in this series support use of the endoscopically placed stent for palliative management of malignant upper gastrointestinal obstruction. PMID- 14519727 TI - The accuracy of guestimates. AB - At times a clinician must describe the size of a lesion when measurement presents difficulties. A 'guestimate' may then be recorded. We tested the accuracy of forensic clinicians (10 physicians and 1 nurse) in guestimating the sizes of ten test objects, comparing their performance with 13 individuals from other professions. The participants were permitted to handle the objects but not to use a measuring device. For three of the objects (a balloon, a scratch and a coin) the entire sample of participants significantly overestimated size, by 13-22%. Both participant groups overestimated the size of a scratch, the clinicians being more accurate but not significantly so. Guestimates should be avoided unless the use of a ruler or tape measure will be against the interests of the client. PMID- 14519728 TI - Prolapsing polyp and disappearing pseudocyst. PMID- 14519729 TI - Disseminated herpes simplex after total skin electron beam radiotherapy for mycosis fungoides. PMID- 14519730 TI - Paraplegia after elective repair of an infrarenal aortic aneurysm. PMID- 14519731 TI - Cutaneous T-cell lymphoma with adrenal insufficiency. PMID- 14519732 TI - Lifelong migraine aura without headache: change of pattern with upper respiratory infection. PMID- 14519733 TI - Combined surgical and endoscopic clearance of small-bowel polyps in Peutz-Jeghers syndrome. PMID- 14519734 TI - The foot that stalled a thousand ships: a controversial case from the 13th century BCE. PMID- 14519735 TI - What should the citizen know about 'science'? PMID- 14519736 TI - Living wills. PMID- 14519737 TI - Living wills. PMID- 14519738 TI - Diagnosis of perforated enterocystoplasty. PMID- 14519739 TI - Medically unexplained symptoms. PMID- 14519740 TI - Patients' memory for medical information. PMID- 14519741 TI - Adjuvant chemotherapy for non-small-cell lung cancer: the end of the beginning. PMID- 14519743 TI - VEGF's Surprise Role in Neurodegenerative Disease. PMID- 14519742 TI - Evidence of efficacy: researchers investigating markers for angiogenesis inhibitors. PMID- 14519744 TI - Despite challenges, cancer prevention and control programs on the rise. PMID- 14519745 TI - Capitalizing on commercialization: should congress revisit the Bayh-Dole Act? PMID- 14519747 TI - Report examines association between cancer and socioeconomic status. PMID- 14519748 TI - Stat bite: Distant-stage prostate cancer diagnoses by ethnicity and socioeconomic status. PMID- 14519749 TI - Comparing survival of a sample to that of a standard population. PMID- 14519750 TI - Enhancing radiotherapy with cyclooxygenase-2 enzyme inhibitors: a rational advance? AB - Results of preclinical studies suggesting that the efficacy of molecular therapies is enhanced when they are combined with radiation have generated a surge of clinical trials combining these modalities. We reviewed the literature to identify the rationale and experimental foundation supporting the use of cyclooxygenase-2 (COX-2) inhibitors with standard radiotherapy regimens in current clinical trials. Radiation affects the ability of cells to divide and proliferate and induces the expression of genes involved in signaling pathways that promote cell survival or trigger cell death. Future advances in radiotherapy will hinge on understanding mechanisms by which radiation-induced transcription of genes governs cell death and survival, the selective control of this process, and the optimal approaches to combining this knowledge with existing therapeutic modalities. COX-2 is expressed in all stages of cancer, and in several cancers its overexpression is associated with poor prognosis. Evidence from clinical and preclinical studies indicates that COX-2-derived prostaglandins participate in carcinogenesis, inflammation, immune response suppression, apoptosis inhibition, angiogenesis, and tumor cell invasion and metastasis. Clinical trial results have demonstrated that selective inhibition of COX-2 can alter the development and the progression of cancer. In animal models, selective inhibition of COX-2 activity is associated with the enhanced radiation sensitivity of tumors without appreciably increasing the effects of radiation on normal tissue, and preclinical evidence suggests that the principal mechanism of radiation potentiation through selective COX-2 inhibition is the direct increase in cellular radiation sensitivity and the direct inhibition of tumor neovascularization. Results of current early-phase studies of non-small-cell lung, esophageal, cervical, and brain cancers will determine whether therapies that combine COX-2 inhibitors and radiation will enter randomized clinical trials. PMID- 14519751 TI - Randomized study of adjuvant chemotherapy for completely resected stage I, II, or IIIA non-small-cell Lung cancer. AB - BACKGROUND: Surgery is the primary treatment for patients with stage I, II, or IIIA non-small-cell lung cancer (NSCLC). However, long-term survival of NSCLC patients after surgery alone is largely unsatisfactory, and the role of adjuvant chemotherapy in patient survival has not yet been established. METHODS: Between January 1994 and January 1999, 1209 patients with stage I, II, or IIIA NSCLC were randomly assigned to receive mitomycin C (8 mg/m2 on day 1), vindesine (3 mg/m2 on days 1 and 8), and cisplatin (100 mg/m2 on day 1) every 3 weeks for three cycles (MVP group; n = 606) or no treatment (control group; n = 603) after complete resection. Randomization was stratified by investigational center, tumor size, lymph-node involvement, and the intention to perform radiotherapy. The primary endpoint was overall survival and secondary endpoints were progression free survival and toxicity associated with adjuvant treatment. Survival curves were analyzed using the log-rank test. All statistical tests were two-sided. RESULTS: After a median follow-up time of 64.5 months, there was no statistically significant difference between the two patient groups in overall survival (hazard ratio = 0.96, 95% confidence interval = 0.81 to 1.13; P =.589) or progression free survival (hazard ratio = 0.89, 95% confidence interval = 0.76 to 1.03; P =.128). Only 69% of patients received the three planned cycles of MVP. Grades 3 and 4 neutropenia occurred in 16% and 12%, respectively, of patients in the MVP arm. Radiotherapy was completed by 65% of patients in the MVP arm and by 82% of patients in the control group. In the multivariable analysis, only disease stage and sex were associated with survival. CONCLUSION: This randomized trial failed to prospectively confirm a statistically significant role for adjuvant chemotherapy in completely resected NSCLC. Given the poor compliance with the MVP regimen used in this study, future studies should explore more effective treatments. PMID- 14519752 TI - Interval carcinomas in the European Randomized Study of Screening for Prostate Cancer (ERSPC)-Rotterdam. AB - BACKGROUND: The interval cancer rate is an important parameter for determining the sensitivity of a screening procedure and the screening interval. We evaluated the time and mechanism of detection and the stage distribution of interval prostate cancers diagnosed during a 4-year screening interval. METHODS: We determined the rate of interval cancers and the sensitivity of the screening protocol (involving prostate-specific antigen, digital rectal and transrectal ultrasound examinations) in a cohort of 17 226 men (8350 on the screened arm, 8876 on the control arm) enrolled consecutively on the European Randomized Study of Screening for Prostate Cancer-Rotterdam. Men on the screened arm received a first screen between October 1993 and December 1996 and a scheduled second screen 4 years later. Prostate cancers detected in men enrolled on the control arm over the same 4-year period and, between screens, in men on the screened arm, were identified by linkage to the Dutch national cancer registry. RESULTS: During the first screen, 412 prostate cancers were detected. During the subsequent 4-year period, 135 cancers were diagnosed in men in the control arm and 25 cancers were diagnosed in men in the screened arm. Seven of the 25 cancers were diagnosed in men who had refused a recommended biopsy at their initial screen. Of the remaining 18 cancers, all were classified as stage T1A-C or T2A and none were poorly differentiated or metastatic. The rate of interval cancers relative to the number of cancers in the control group was 18.5% (25/135), or 13.3% (18/135), if the seven who refused an initial biopsy were excluded. The sensitivity of the screening protocol was 79.8% when considering all 25 interval cancers and 85.5% when considering 18 interval cancers. CONCLUSION: The interval cancer rate with a 4-year screening interval was low, confirming that the screening procedure has a high sensitivity and that the 4-year screening interval is reasonable. PMID- 14519753 TI - Obesity, tamoxifen use, and outcomes in women with estrogen receptor-positive early-stage breast cancer. AB - BACKGROUND: Obesity is associated with both increased breast cancer risk and poorer prognosis after disease onset. However, little is known about the effect of obesity on treatment efficacy. We evaluated the association of obesity with outcomes and with tamoxifen efficacy in women with early-stage, hormone responsive breast cancer participating in a multicenter cancer cooperative group clinical trial. METHODS: The cohort consisted of 3385 women enrolled in National Surgical Adjuvant Breast and Bowel Project (NSABP) protocol B-14, a randomized, placebo-controlled trial evaluating tamoxifen for lymph node-negative, estrogen receptor (ER)-positive breast cancer. Hazards of breast cancer recurrence, contralateral breast tumors, other new primary cancers, and several mortality endpoints were evaluated in relation to body mass index (BMI), using statistical modeling to adjust for other prognostic factors. Median follow-up time was 166 months. All statistical tests were two-sided. RESULTS: The hazard of breast cancer recurrence was the same among obese (BMI > or =30.0 kg/m2) women as compared with underweight and normal-weight women (BMI <25.0; hazard ratio [HR] = 0.98, 95% confidence interval [CI] = 0.80 to 1.18). Contralateral breast cancer hazard was higher in obese women than in underweight/normal-weight women (HR = 1.58, 95% CI = 1.10 to 2.25), as was the risk of other primary cancers (HR = 1.62, 95% CI = 1.16 to 2.24). Compared with normal-weight women, obese women had greater all-cause mortality (HR = 1.31, 95% CI = 1.12 to 1.54) and greater risk of deaths due to causes unrelated to breast cancer (HR = 1.49, 95% CI = 1.15 to 1.92). Breast cancer mortality was not statistically significantly increased for obese women (HR = 1.20, 95% CI = 0.97 to 1.49). Tamoxifen reduced breast cancer recurrence and mortality, regardless of BMI. CONCLUSIONS: For women with lymph node-negative, ER-positive breast cancer, obesity was not associated with a material increase in recurrence risk or a change in tamoxifen efficacy. However, because obesity was associated with increased risks of contralateral breast cancer, of other primary cancers, and of overall mortality, it may influence long term outcomes for breast cancer survivors. PMID- 14519754 TI - Selenium supplementation and secondary prevention of nonmelanoma skin cancer in a randomized trial. AB - The Nutritional Prevention of Cancer Trial was a double-blind, randomized, placebo-controlled clinical trial designed to test whether selenium as selenized yeast (200 microg daily) could prevent nonmelanoma skin cancer among 1312 patients from the Eastern United States who had previously had this disease. Results from September 15, 1983, through December 31, 1993, showed no association between treatment and the incidence of basal and squamous cell carcinomas of the skin. This report summarizes the entire blinded treatment period, which ended on January 31, 1996. The association between treatment and time to first nonmelanoma skin cancer diagnosis and between treatment and time to multiple skin tumors overall and within subgroups, defined by baseline characteristics, was evaluated. Although results through the entire blinded period continued to show that selenium supplementation was not statistically significantly associated with the risk of basal cell carcinoma (hazard ratio [HR] = 1.09, 95% confidence interval [CI] = 0.94 to 1.26), selenium supplementation was associated with statistically significantly elevated risk of squamous cell carcinoma (HR = 1.25, 95% CI = 1.03 to 1.51) and of total nonmelanoma skin cancer (HR = 1.17, 95% CI = 1.02 to 1.34). Results from the Nutritional Prevention of Cancer Trial conducted among individuals at high risk of nonmelanoma skin cancer continue to demonstrate that selenium supplementation is ineffective at preventing basal cell carcinoma and that it increases the risk of squamous cell carcinoma and total nonmelanoma skin cancer. PMID- 14519755 TI - Germline BRCA1 mutations and a basal epithelial phenotype in breast cancer. AB - A basal epithelial phenotype is found in not more than 15% of all invasive breast cancers. Microarray studies have shown that this phenotype is associated with breast cancers that express neither estrogen receptor (ER) nor erbB-2 (HER2/neu) (i.e., ER/erbB-2-negative tumors). The ER/erbB-2- negative phenotype is also found in breast cancers occurring in BRCA1 mutation carriers (i.e., BRCA1-related breast cancers). We tested the hypothesis that BRCA1-related breast cancers are more likely than non-BRCA1/ 2-related breast cancer to express a basal epithelial phenotype. Among 292 breast cancer specimens previously analyzed for ER, erbB-2, p53, and germline mutations in BRCA1 and BRCA2, we identified 76 that did not overexpress ER or erbB-2. Of the 72 specimens with sufficient material for testing, 40 expressed stratified epithelial cytokeratin 5 and/or 6 (5/6). In univariate analysis, the expression of cytokeratin 5/6 was statistically significantly associated with BRCA1-related breast cancers (odds ratio = 9.0, 95% confidence interval = 1.9 to 43; P =.002, two-sided Fisher's exact test). Thus, germline BRCA1 mutations appear to be associated with a distinctive breast cancer phenotype. PMID- 14519756 TI - Polymorphisms in XRCC1 and glutathione S-transferase genes and hepatitis B related hepatocellular carcinoma. AB - Chronic infection with hepatitis B virus (HBV) causes DNA damage. An arginine (Arg)-to-glutamine (Gln) polymorphism at codon 399 in the XRCC1 gene is putatively associated with DNA damage. In a case-control study of 577 HBV surface antigen carriers with hepatocellular carcinoma (HCC) and 389 HBV carrier control subjects, we investigated the association between this polymorphism and the risk of HCC and assessed whether this association varied with glutathione S transferase (GST) status; GSTs are involved in carcinogen metabolism. All statistical tests were two-sided. The XRCC1 Gln allele was associated with a dose dependent increased risk of early-onset HCC (<50 years) but not with the risk of late-onset HCC (P(trend) =.01). The GSTT1-null genotype alone did not affect risk, but the GSTM1-null genotype was associated with a decreased risk for early onset HCC. Various combinations of GSTM1 and GSTT1 genotypes differentially modified the association of XRCC1 with HCC (P(interaction) =.005); e.g., for individuals with the GSTT1-null/GSTM1-present genotype, the risk of HCC was greater for those with the Gln/Gln genotype (odds ratio = 8.07, 95% confidence interval = 1.67 to 38.93) than for those with the Arg/Arg genotype. Thus, GST status appears to affect the risk of HCC associated with this XRCC1 polymorphism. PMID- 14519757 TI - The role of the combination of IL-2 and TGF-beta or IL-10 in the generation and function of CD4+ CD25+ and CD8+ regulatory T cell subsets. AB - Recently, considerable attention has been focused on thymus-derived CD4+ regulatory T cells that constitutively express CD25 and have a contact-dependent, cytokine-independent mechanism in vitro. However, peripheral CD4+ and CD8+ T cells can also be induced to become regulatory T cells. Here we review our studies using the combination of IL-2 and transforming growth factor beta (TGF beta) to generate regulatory T cell subsets ex vivo, and the work of others using IL-10 to induce suppressive activity. Under certain conditions, the autocrine effects of TGF-beta and IL-10 induce peripheral T cells to produce immunosuppressive levels of each of these cytokines. This effect of TGF-beta is IL-2 dependent. Under other conditions IL-2 and TGF-beta can induce CD4+ cells to develop potent contact-dependent, cytokine-independent regulatory activity. At present, there is considerable confusion concerning the mechanism of action of CD4+ CD25+ cells because cytokine-producing regulatory T cells generated in the periphery can express CD25 and other markers displayed by naturally occurring, thymus-derived regulatory T cells. We, therefore, propose a nomenclature that identifies thymus-derived and peripheral regulatory cells, and that also differentiates T regulatory cells from T helper cells. Because T regulatory cells broadly control T helper cell reactivity, the mechanisms that control regulatory cell function are also reviewed. Finally, the potential use of regulatory T cells generated ex vivo as an adoptive immunotherapy for certain autoimmune diseases, to prevent organ graft rejection, or to prevent pathologic host responses to infectious agents is discussed. PMID- 14519758 TI - Copper chelation delays the onset of prion disease. AB - The prion protein (PrP) binds copper and under some conditions copper can facilitate its folding into a more protease resistant form. Hence, copper levels may influence the infectivity of the scrapie form of prion protein (PrPSc). To determine the feasibility of copper-targeted therapy for prion disease, we treated mice with a copper chelator, D-(-)-penicillamine (D-PEN), starting immediately following intraperitoneal scrapie inoculation. D-PEN delayed the onset of prion disease in the mice by about 11 days (p = 0.002), and reduced copper levels in brain by 29% (p < 0.01) and in blood by 22% (p = 0.03) compared with control animals. Levels of other metals were not significantly altered in the blood or brain. Modest correlation was observed between incubation period and levels of copper in brain (p = 0.08) or blood (p = 0.04), indicating that copper levels are only one of many factors that influence the rate of progression of prion disease. In vitro, copper dose-dependently enhanced the proteinase K resistance of the prion protein, and this effect was counteracted in a dose dependent manner by co-incubation with D-PEN. Overall, these findings indicate that copper levels can influence the conformational state of PrP, thereby enhancing its infectivity, and this effect can be attenuated by chelator-based therapy. PMID- 14519759 TI - The human dopamine transporter forms a tetramer in the plasma membrane: cross linking of a cysteine in the fourth transmembrane segment is sensitive to cocaine analogs. AB - Using cysteine cross-linking, we demonstrated previously that the dopamine transporter (DAT) is at least a homodimer, with the extracellular end of transmembrane segment (TM) 6 at a symmetrical dimer interface. We have now explored the possibility that DAT exists as a higher order oligomer in the plasma membrane. Cysteine cross-linking of wild type DAT resulted in bands on SDS-PAGE consistent with dimer, trimer, and tetramer, suggesting that DAT forms a tetramer in the plasma membrane. A cysteine-depleted DAT (CD-DAT) into which only Cys243 or Cys306 was reintroduced was cross-linked to dimer, suggesting that these endogenous cysteines in TM4 and TM6, respectively, were cross-linked at a symmetrical dimer interface. Reintroduction of both Cys243 and Cys306 into CD-DAT led to a pattern of cross-linking indistinguishable from that of wild type, with dimer, trimer, and tetramer bands. This indicated that the TM4 interface and the TM6 interface are distinct and further suggested that DAT may exist in the plasma membrane as a dimer of dimers, with two symmetrical homodimer interfaces. The cocaine analog MFZ 2-12 and other DAT inhibitors, including benztropine and mazindol, protected Cys243 against cross-linking. In contrast, two substrates of DAT, dopamine and tyramine, did not significantly impact cross-linking. We propose that the impairment of cross-linking produced by the inhibitors results from a conformational change at the TM4 interface, further demonstrating that these compounds are not neutral blockers but by themselves have effects on the structure of the transporter. PMID- 14519760 TI - Generation of a recombinant, membrane-targeted form of the complement regulator CD59: activity in vitro and in vivo. AB - Inappropriate activation of complement contributes to pathology in diverse inflammatory diseases. Soluble recombinant forms of the natural cell membrane regulators of complement are effective in animal models and some human diseases. However, their use is limited for reasons related to cost, short half lives, and propensity to cause unwanted systemic effects. Some of these limitations may be overcome by use of bacterial expression systems, specific targeting moieties, and judicious choice of regulator. Here we describe the application of these strategies to the generation of a membrane-targeted form of CD59. A recombinant soluble form of rat CD59, comprising the first 71 residues of the mature protein and missing the membrane-anchoring signal, was expressed in bacteria, purified, and refolded in a fully active form. The protein was coupled through its carboxyl terminus to a short, synthetic address tag that confers membrane binding activity. Attachment of the membrane address tag markedly increased complement inhibitory activity assessed in vitro in hemolysis assays. Intra-articular administration of the tagged agent markedly suppressed disease in a model of rheumatoid arthritis in Lewis rats. This novel type of agent, termed sCD59 APT542, offers for the first time the prospect of efficient and specific inhibition of membrane attack complex activity in vivo. PMID- 14519761 TI - Tumor necrosis factor alpha modulates airway smooth muscle function via the autocrine action of interferon beta. AB - Current evidence suggests that tumor necrosis factor alpha (TNFalpha) and the family of interferons (IFNs) synergistically regulate many cellular responses that are believed to be critical in chronic inflammatory diseases, although the underlying mechanisms of such interaction are complex, cell-specific, and not completely understood. In this study, TNFalpha in a time-dependent manner activated both janus tyrosine kinase 1 and Tyk2 tyrosine kinase and increased the nuclear translocation of interferon-regulatory factor-1, STAT1, and STAT2 in human airway smooth muscle cells. In cells transfected with a luciferase reporter, TNFalpha stimulated gamma-activated site-dependent gene transcription in a time- and concentration-dependent manner. Using neutralizing antibodies to IFNbeta and TNFalpha receptor 1, we show that TNFalpha-induced secretion of IFNbeta mediated gamma-activated site-dependent gene expression via activation of TNFalpha receptor 1. In addition, neutralizing antibody to IFNbeta also completely abrogated the activation of interferon stimulation response element dependent gene transcription induced by TNFalpha. Secreted IFNbeta acted as a negative regulator of TNFalpha-induced interleukin-6 expression, while IFNbeta augmented TNFalpha-induced RANTES (regulated on activation normal T cell expressed and secreted) secretion but had little effect on TNFalpha-induced intercellular adhesion molecule-1 expression. Furthermore TNFalpha, a modest airway smooth muscle mitogen, markedly induced DNA synthesis when cells were treated with neutralizing anti-IFNbeta. Together these data show that TNFalpha, via the autocrine action of IFNbeta, differentially regulates the expression of proinflammatory genes and DNA synthesis. PMID- 14519762 TI - Immune activation of NF-kappaB and JNK requires Drosophila TAK1. AB - Stimulation of the Drosophila immune response activates NF-kappaB and JNK signaling pathways. For example, infection by Gram-negative bacteria induces the Imd signaling pathway, leading to the activation of the NF-kappaB-like transcription factor Relish and the expression of a battery of genes encoding antimicrobial peptides. Bacterial infection also activates the JNK pathway, but the role of this pathway in the immune response has not yet been established. Genetic experiments suggest that the Drosophila homolog of the mammalian MAPK kinase kinase, TAK1 (transforming growth factor beta-activated kinase 1), activates both the JNK and NF-kappaB pathways following immune stimulation. In this report, we demonstrate that Drosophila TAK1 functions as both the Drosophila IkappaB kinase-activating kinase and the JNK kinase-activating kinase. However, we found that JNK signaling is not required for antimicrobial peptide gene expression but is required for the activation of other immune inducible genes, including Punch, sulfated, and malvolio. Thus, JNK signaling appears to play an important role in the cellular immune response and the stress response. PMID- 14519763 TI - Chemoenzymatic synthesis of classical and non-classical anticoagulant heparan sulfate polysaccharides. AB - Heparan sulfate (HS) polysaccharides interact with numerous proteins at the cell surface and orchestrate many different biological functions. Though many functions of HS are well established, only a few specific structures can be attributed to HS functions. The extreme diversity of HS makes chemical synthesis of specific bioactive HS structures a cumbersome and tedious undertaking that requires laborious and careful functional group manipulations. Now that many of the enzymes involved in HS biosynthesis are characterized, we show in this study how one can rapidly and easily assemble bioactive HS structures with a set of cloned enzymes. We have demonstrated the feasibility of this new approach to rapidly assemble antithrombin III-binding classical and non-classical anticoagulant polysaccharide structures for the first time. PMID- 14519764 TI - Group I metabotropic glutamate receptors bind to protein phosphatase 1C. Mapping and modeling of interacting sequences. AB - The modulation of neurotransmitter receptors by kinases and phosphatases represents a key mechanism in controlling synaptic signal transduction. However, molecular determinants involved in the specific targeting and interactions of these enzymes are largely unknown. Here, we identified both catalytic gamma isoforms of protein phosphatase 1C (PP1gamma1 and PP1gamma2) as binding partners of the group I metabotropic glutamate receptors type 1a, 5a, and 5b in yeast cells and pull-down assays, using recombinant and native protein preparations. The tissue distribution of interacting proteins was compared, and protein phosphatase 1C was detected in dendrites of retinal bipolar cells expressing the respective interacting glutamate receptors. We mapped interacting domains within binding partners and identified five amino acids in the intracellular C termini of the metabotropic glutamate receptors type 1a, 5a, 5b, and 7b being both necessary and sufficient to bind protein phosphatase 1C. Furthermore, we show a dose-dependent competition of these C termini in binding the enzyme. Based on our data, we investigated the structure of the identified amino acids bound to protein phosphatase 1C by homology-based molecular modeling. In summary, these results provide a molecular description of the interaction between protein phosphatase 1C and metabotropic glutamate receptors and thereby increase our understanding of glutamatergic signal transduction. PMID- 14519765 TI - TIR-containing adapter molecule (TICAM)-2, a bridging adapter recruiting to toll like receptor 4 TICAM-1 that induces interferon-beta. AB - Lipopolysaccharide (LPS) is an agonist for Toll-like receptor (TLR) 4 and expresses many genes including NF-kappaB- and interferon regulatory factor (IRF) 3/IFN-inducible genes in macrophages and dendritic cells (DCs). TICAM-1/TRIF was identified as an adapter that facilitates activation of IRF-3 followed by expression of interferon (IFN)-beta genes in TLR3 signaling, but TICAM-1 does not directly bind TLR4. Although MyD88 and Mal/TIRAP adapters functions downstream of TLR4, DC maturation and IFN-beta induction are independent of MyD88 and Mal/TIRAP. In this investigation, we report the identification of a novel adapter, TICAM-2, that physically bridges TLR4 and TICAM-1 and functionally transmits LPS-TLR4 signaling to TICAM-1, which in turn activates IRF-3. In its structural features, TICAM-2 resembled Mal/TIRAP, an adapter that links TLR2/4 and MyD88. However, TICAM-2 per se exhibited minimal ability to activate NF kappaB and the IFN-beta promoter. Hence, in LPS signaling TLR4 recruits two types of adapters, TIRAP and TICAM-2, to its cytoplasmic domain that are indirectly connected to two effective adapters, MyD88 and TICAM-1, respectively. We conclude that for LPS-TLR4-mediated activation of IFN-beta, the adapter complex of TICAM-2 and TICAM-1 plays a crucial role. This results in the construction of MyD88 dependent and -independent pathways separately downstream of the two distinct adapters. PMID- 14519766 TI - Differential roles of C-terminal activation motifs in the establishment of Stat6 transcriptional specificity. AB - Members of the Stat transcription factor family are specifically activated by cytokines, and each Stat mediates its biological effects through the trans activation of a unique profile of target genes. This specificity is achieved even when Stat proteins mediating opposite transcriptional effects bind to the same palindromic Stat sites in target genes. We show here that the non-conserved sequences of Stat transcription activation domains (TADs) contribute to specificity in promoter activation. Chimeric proteins in which the Stat6 TAD was replaced by that from Stat1alpha or Stat5 exhibited normal interleukin-4 inducible DNA binding activity, but at best modest trans-activation of reporters containing Stat6 binding sites, and a failure to activate the endogenous CD23 promoter in primary B cells. The p160 coactivator nuclear coactivator-1 (Src-1) was specifically recruited by and coactivated Stat6 but not the chimeric Stat6 molecules. Strikingly, transcriptional responses exhibited distinct requirements for the nuclear coactivator-1 interaction motif of the Stat6 C terminus. Together, these findings indicate that the Stat6 TAD contributes to promoter specificity by the differential recruitment of and requirement for a p160-class coactivator. PMID- 14519767 TI - Age-related changes in the transcriptional profile of mouse RPE/choroid. AB - To evaluate the age-related changes in gene expression occurring in the complex of retinal pigmented epithelium, Bruch's membrane, and choroid (RPE/choroid), we examined the gene expression profiles of young adult (2 mo) and old (24 mo) male C57BL/6 mice. cDNA probe sets from individual animals were synthesized using total RNA isolated from the RPE/choroid of each animal. Probes were amplified using the Clontech SMART system, radioactively labeled, and hybridized to two different Clontech Atlas mouse cDNA arrays. From each age group, three independent triplicates were hybridized to the arrays. Statistical analyses were performed using the Significance Analysis of Microarrays program (SAM version 1.13; Stanford University). Selected array results were confirmed by semi quantitative RT-PCR analysis. Of 2,340 genes represented on the arrays, approximately 60% were expressed in young and/or old mouse RPE/choroid. A moderate fraction (12%) of all expressed genes exhibited a statistically significant change in expression with age. Of these 150 genes, all but two, HMG14 and carboxypeptidase E, were upregulated with age. Many of these upregulated genes can be grouped into several broad functional categories: immune response, proteases and protease inhibitors, stress response, and neovascularization. RT PCR results from six of six genes examined confirmed the differential change in expression with age of these genes. Our study provides likely candidate genes to further study their role in the development of age-related macular degeneration and other aging diseases affecting the RPE/choroid. PMID- 14519768 TI - Rice globular embryo 4 (gle4) mutant is defective in radial pattern formation during embryogenesis. AB - In higher plants, the main elements of the fundamental body plan, the apical basal and radial patterns, are established during embryogenesis. We have isolated several globular embryo (gle) mutants of rice that fail to develop any embryonic organs. We expected that these gle mutants might include mutants defective in their radial pattern formation ability. We developed two markers specifically staining the L2 and L3 layers (OsSCR and OsPNH1, respectively) and characterized the gle mutants by using these markers in addition to the already developed markers Roc1 (marker for the L1 layer), Ramy1A (marker of the L1 layer of the epithelium), and OSH1 (marker of the apical region). One of the gle mutants, gle4, expressed Roc1 and Ramy1A at the normal positions, but other markers exhibited an abnormal expression pattern; that is, both OsPNH1 and OsSCR were expressed in the central region of the embryo and OSH1 expression was not observed. Calli from the gle4 epithelium regenerated plants with abnormal morphologies. These results indicate that the GLE4 gene is involved in radial pattern formation during rice embryogenesis to differentiate the L2 and L3 layers, but is not involved in the establishment of the L1 layer or in the formation of embryonic organs. PMID- 14519769 TI - Hyper-expression of small nucleolar RNAs (snoRNAs) in female inflorescences of hazelnut (Corylus avellana L.) supports rRNA aggregation in vitro. AB - Under certain in vitro (salt and temperature) conditions rRNA aggregation occurs in female inflorescences but not in leaves or pollen RNA preparations from hazelnut (Corylus avellana L.), a species of economic interest. This paper describes experiments addressing an explanation of this phenomenon. The experiments demonstrate that: (i) trans-acting factors induce rRNA aggregate formation in female inflorescences RNA preparations; (ii) these factors support aggregation also of heterologous rRNA; (iii) aggregation is a function of temperature pre-treatment of rRNA and not of source 18S rRNA; (iv) the factors inducing rRNA aggregates are sensitive to RNase; (v) antisense small nucleolar RNAs (snoRNAs) participate in rRNA aggregate formation. snoRNAs are involved in pre-rRNA spacer cleavages, and are required for the two most common types of rRNA modifications: 2'-O-ribose methylation and pseudouridylation. Even though it is questionable whether rRNA aggregation really happens in female inflorescence in vivo, the phenomenon observed in vitro may reflect the abundance of snoRNAs in these reproductive structures. In fact the level of accumulation of three tested snoRNAs, R1, U14 and U3, is much higher in female inflorescence than in leaves or pollen of hazelnut. This finding opens the possibility of studying the role of snoRNAs in tissue development in plants. PMID- 14519770 TI - Decay kinetics of autogenously regulated CGS1 mRNA that codes for cystathionine gamma-synthase in Arabidopsis thaliana. AB - Cystathionine gamma-synthase (CGS) catalyses the first committed step in methionine (Met) biosynthesis in higher plants. Stability of CGS1 mRNA encoding CGS in Arabidopsis thaliana is regulated by negative feedback in response to Met application and the amino acid sequence of CGS itself acts in cis in this regulation. It is proposed that the regulation occurs during translation when the nascent polypeptide of CGS and its mRNA are in close proximity. This model predicts that inhibition of translation abolishes the regulation. To test this, we analysed the effect of translation inhibitor cycloheximide on the CGS1 mRNA decay. The half-life of CGS1 mRNA after the addition of transcription inhibitor actinomycin D in the absence and presence of 1 mM Met was 154+/-11 min and 81+/-5 min, respectively. Simultaneous addition of actinomycin D and cycloheximide stabilized CGS1 mRNA both in the presence and absence of Met, as essentially no decrease of CGS1 mRNA was observed. Moreover, cycloheximide treatment inhibited production of the truncated CGS1 RNA species, a possible degradation intermediate. These results indicated that inhibition of translation abolishes the CGS1 mRNA-specific decay process. Kinetic analyses indicated that about half the CGS1 mRNA is destined to CGS1 mRNA-specific decay when 1 mM Met was applied. PMID- 14519771 TI - Alteration of citrate metabolism in cluster roots of white lupin. AB - Organic acid excretion plays a key role in the superior P(i)-acquisition of barely soluble inorganic P sources from soils. Seedlings of white lupin (Lupinus albus L.) grown for 37 d in -P nutrient solution showed typical -P symptoms, such as low P content, increased root/shoot ratio and the development of cluster roots which released large amounts of citrate. Citrate concentration in the cluster roots was 21.5 micro mol (g FW)(-1), which corresponded to a 4.3- and 2.6-fold increase of +P and -P root apexes, respectively. Cluster roots possessed higher phosphoenolpyruvate carboxylase and phosphoenolpyruvate phosphatase activity than those in +P root apexes, which could result in increasing the supply of substrate for citrate synthase. On the other hand, the cytosolic pathway which converts citrate to 2-oxoglutarate consists of aconitase and NADP-specific isocitrate dehydrogenase activity that was lower in the cluster roots than in +P root apexes, and may contribute to citrate accumulation. Thus, metabolic balance with these alterations would play an important role in increasing citrate concentration in the cluster roots. The molecular characterization of NADP specific isocitrate dehydrogenase indicated that the cytosolic isoenzyme functions as a hetero-dimer, and that the activity would be regulated by the transcript levels for both isoforms. PMID- 14519772 TI - New fava bean guard cell signaling mutant impaired in ABA-induced stomatal closure. AB - We isolated a mutant from Vicia faba L. cv. House Ryousai. It wilts easily under strong light and high temperature conditions, suggesting that its stomatal movement may be disturbed. We determined responses of mutant guard cells to some environmental stimuli. Mutant guard cells demonstrated an impaired ability to respond to ABA in 0.1 mM CaCl(2) and stomata did not close in the presence of up to 1 mM ABA, whereas wild-type stomata closed when exposed to 10 micro M ABA. Elevating external Ca(2+) caused a similar degree of stomatal closure in the wild type and the mutant. A high concentration of CO(2) (700 micro l liter(-1)) induced stomatal closure in the wild type, but not in the mutant. On the basis of these results, we propose the working hypothesis that the mutation occurs in the region downstream of CO(2) and ABA sensing and in the region upstream of Ca(2+) elevation. The mutant is named fia (fava bean impaired in ABA-induced stomatal closure). PMID- 14519773 TI - Exclusion of ribulose-1,5-bisphosphate carboxylase/oxygenase from chloroplasts by specific bodies in naturally senescing leaves of wheat. AB - Immunocytochemical electron-microscopic observation indicated that ribulose-1,5 bisphosphate carboxylase/oxygenase (Rubisco, EC 4.1.1.39) and/or its degradation products are localized in small spherical bodies having a diameter of 0.4-1.2 micro m in naturally senescing leaves of wheat (Triticum aestivum L.). These Rubisco-containing bodies (RCBs) were found in the cytoplasm and in the vacuole. RCBs contained another stromal protein, chloroplastic glutamine synthetase, but not thylakoid proteins. Ultrastructural analysis suggested that RCBs had double membranes, which seemed to be derived from the chloroplast envelope, and that RCBs were further surrounded by the other membrane structures in the cytoplasm. The appearance of RCBs was the most remarkable when the amount of Rubisco started to decrease at the early phase of leaf senescence. These results suggest that RCBs might be involved in the degradation process of Rubisco outside of chloroplasts during leaf senescence. PMID- 14519774 TI - The leaf-order-dependent enhancement of freezing tolerance in cold-acclimated Arabidopsis rosettes is not correlated with the transcript levels of the cold inducible transcription factors of CBF/DREB1. AB - The central part of cold-acclimated rosettes of Arabidopsis thaliana L. (ecotype Columbia) survived freezing at lower temperatures better than did those at the rosette periphery. Electrolyte-leakage tests with detached leaves verified that freezing tolerance in central (or young) leaves increased faster and to a greater extent than in peripheral (or aged and mature) leaves at 2 degrees C. Cold induced accumulation of sugars could partly account for the leaf-order-dependent enhancement of freezing tolerance after 1 d at 2 degrees C, whereas the role of proline remains to be determined. Cold-induced accumulation of the transcripts of stress-inducible CBF/DREB1 transcription factors apparently disagreed with the observed difference in the freezing tolerance in different leaf orders. However, the levels of COR78/RD29A transcripts were almost the same between different leaf orders after 1-3 d at 2 degrees C, and COR78/RD29A content per total leaf protein was similar between different leaf orders after 7 d at 2 degrees C. Thus, cold induced accumulation of COR78/RD29A does not seem to account for the observed difference in freezing tolerance in different leaf orders. Although further studies are required for comprehensive understanding of the phenomenon, the present work does provide an important and interesting physiological aspect in our understanding of the freezing tolerance in plants. PMID- 14519775 TI - Ethylene promotes the induction by auxin of the cortical microtubule randomization required for low-pH-induced root hair initiation in lettuce (Lactuca sativa L.) seedlings. AB - Transverse cortical microtubule (CMT) arrays in lettuce root epidermal cells randomize soon after a shift from pH 6.0 to pH 4.0, and this randomization is essential for root hair initiation. We investigated the hormonal regulation of CMT randomization. At pH 4.0, 1 micro M of the auxin competitive inhibitor 2-(p chlorophenoxy)-2-methylpropionic acid (PCIB), 0.1 micro M of the ethylene biosynthesis inhibitor aminoethoxyvinylglycine (AVG) or 0.1 micro M of the ethylene action inhibitor Ag(+) suppressed CMT randomization and root hair initiation. At pH 6.0, addition of 0.1 micro M indole-3-acetic acid (IAA) or 1 micro M of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) induced CMT randomization and root hair initiation. Culturing with 0.1 micro M IAA plus 0.1 micro M AVG, or 1 micro M ACC plus 1 micro M PCIB also induced these phenomena. ACC (1 micro M) plus 100 micro M PCIB inhibited CMT randomization and root hair initiation, but 1 micro M AVG with 0.1 micro M Ag(+) and 0.1 micro M IAA induced them. These results suggest that auxin is essential for CMT randomization. As a higher concentration of PCIB was required to suppress CMT randomization when ACC was added, the greater amount of ethylene produced at pH 4.0 may promote the induction by auxin of CMT randomization in hair-forming cells. PMID- 14519776 TI - Examination of the cytoplasmic DNA in male reproductive cells to determine the potential for cytoplasmic inheritance in 295 angiosperm species. AB - Mature pollen grains of 295 angiosperm species were screened by epifluorescence microscopy for a marker that denotes the mode of cytoplasmic inheritance. We used the DNA fluorochrome DAPI (4',6-diamidino-2-phenylindole) for pollen cell staining. The presence or absence of fluorescence of cytoplasmic DNA in the generative cell or sperm cells was examined in each species. The species examined represented 254 genera and 98 families, and 40 of these families had not been previously studied in this regard. The cytoplasmic DNA of the generative cell or sperm cells did not fluoresce in 81% of the species examined, from 83% of the genera and 87% of the families examined, indicating the potential for maternal cytoplasmic inheritance in these species. In contrast, the male reproductive cells of 19% of the species, from 17% of the genera and 26% of the families examined, displayed fluorescence of the cytoplasmic DNA, indicating the potential for biparental cytoplasmic inheritance in these species. The results revealed the potential for biparental cytoplasmic inheritance in several species in which the inheritance mode was previously unknown, including plants in the Bignoniaceae, Cornaceae, Cruciferae (Brassicaceae), Cyperaceae, Dipsacaceae, Hydrocharitaceae, Papaveraceae, Portulacaceae, Tiliaceae, Valerianaceae, and Zingiberaceae. Electron microscopy revealed that the sperm cells of Portulaca grandiflora contain both plastid and mitochondrial DNA. However, in the generative cells of Musella lasiocarpa, the mitochondria contain DNA, but the plastids do not. These data provide a foundation for further studies of cytoplasmic inheritance in angiosperms. PMID- 14519777 TI - Differences between maize and rice in N-use efficiency for photosynthesis and protein allocation. AB - The N-use efficiency for photosynthesis was higher in a C(4) plant, maize, than in a C(3) plant, rice, including rbcS antisense rice with optimal ribulose-1,5 bisphosphate carboxylase (Rubisco) content for CO(2)-saturated photosynthesis, even when photosynthesis was measured under saturating CO(2) conditions. The N cost for the C(4) cycle enzymes in maize was not large, and the lower amount of Rubisco allowed a greater N investment in the thylakoid components. This greater content of the thylakoid components as well as the CO(2) concentrating mechanism may support higher N-use efficiency for photosynthesis in maize. PMID- 14519778 TI - Studies on alkaline band formation in Chara corallina: ameliorating effect of Ca2+ on inhibition induced by osmotic shock. AB - Although the decrease in cell turgor by application of sorbitol to the external medium did not inhibit the alkaline band formation in Chara corallina, recovery of normal turgor severely inhibited it. Alkaline-loading analysis suggested that the inhibition of alkaline band formation was caused by inhibition of HCO(3)(-) influx but not that of OH(-) efflux. In the presence of 10 mM CaCl(2), the capacity of alkaline band formation was maintained during osmotic treatment. Cells could not form alkaline bands, when plasmolysis was induced by application of sorbitol at a higher concentration. Addition of 10 mM CaCl(2) could ameliorate the inhibition caused by plasmolyis. PMID- 14519779 TI - A short history of nutritional science: part 3 (1912-1944). PMID- 14519780 TI - Nutritional genomics: implications for companion animals. AB - This is an exciting time for biological scientists as the "omics" era continues to evolve and shape the way science is understood and conducted. As genome sequencing of the human comes to a close, other mammals are in line to be sequenced. Along with pigs and cows, dogs are now on the high priority list for sequencing, and cats may soon follow suit. Until sequence data are available, genetic maps may be used to reveal important physical characteristics of a genome. Genome mapping is important in identifying gene placement, but gives little information regarding function. Therefore, functional genomics, including the global analysis of RNA and protein expression, protein localization and protein-protein interactions will emerge as important areas of study. The major use of the dog and cat genome maps hitherto has been for the study of human and veterinary medicine. These powerful resources also can be applied to the field of nutritional genomics and proteomics, enhancing our understanding of metabolism and optimizing companion animal nutritional and health status. Genomics has begun to be applied to nutritional research, but issues specifically relevant to companion animals have not been elucidated thus far. The study of genomics and proteomics will be crucial in areas such as nutrient requirement determination, disease prevention and treatment, functional ingredient testing and others. Nutritional genomics and proteomics will definitely play a vital role in the future of pet foods. PMID- 14519781 TI - Conjugated linoleic acid in humans: regulation of adiposity and insulin sensitivity. AB - Conjugated linoleic acid (CLA) isomers, a group of positional and geometric isomers of linoleic acid [18:2(n-6)], have been studied extensively due to their ability to modulate cancer, atherosclerosis, obesity, immune function and diabetes in a variety of experimental models. The purpose of this review was to examine CLA's isomer-specific regulation of adiposity and insulin sensitivity in humans and in cultures of human adipocytes. It has been clearly demonstrated that specific CLA isomers or a crude mixture of CLA isomers prevent the development of obesity in certain rodent and pig models. This has been attributed mainly to trans-10, cis-12 CLA, both in vivo and in vitro. However, CLA's ability to modulate human obesity remains controversial because data from clinical trials using mixed isomers are conflicting. In support of some studies in humans, our group demonstrated that trans-10, cis-12 CLA prevents triglyceride (TG) accumulation in primary cultures of differentiating human preadipocytes. In contrast, cis-9, trans-11 CLA increases TG content. Closer examination has revealed that CLA's antiadipogenic actions are due, at least in part, to regulation of glucose and fatty acid uptake and metabolism. This review presents our current understanding of potential isomer-specific mechanisms by which CLA reduces human adiposity and insulin sensitivity. PMID- 14519782 TI - Vitamin C inhibits lipid oxidation in human HDL. AB - HDL are susceptible to oxidation, which affects their cardioprotective properties. Although several studies have reported inhibition of HDL oxidation by vitamin E, none has determined the potential protective effect of vitamin C, another important blood antioxidant. We investigated whether vitamin C protects HDL from oxidation by incubating HDL (0.2 g of protein/L) at 37 degrees C with cupric (Cu2+) ions (10 micromol/L) in the absence (control) or presence of vitamin C (20-200 micromol/L). In the absence of vitamin C, lipid oxidation in HDL began immediately and proceeded rapidly. Cholesteryl linoleate declined to a minimum, whereas lipid oxidation products (lipid dienes and TBARS) increased to near-maximal levels within 1 h. Vitamin C (50-200 micromol/L) retarded initiation of lipid oxidation for at least 4 h under the same conditions. The ability of vitamin C to preserve the cardioprotective antioxidant function of HDL was also assessed. HDL (0.5 g of protein/L) preincubated with Cu2+ (10 micromol/L) for 2 h in the absence of vitamin C lost antioxidant activity (45.4 +/- 6.2% inhibition of LDL oxidation compared with 93.2 +/- 3.6% for native HDL, P < 0.05). The addition of vitamin C (50-200 micromol/L) during preincubation of HDL with Cu2+, however, resulted in no significant loss of HDL antioxidant activity (77.3 +/- 0.3 to 89.8 +/- 5.4% inhibition of LDL oxidation, P > 0.05 compared with native HDL). Our results demonstrate that vitamin C inhibits lipid oxidation in HDL and preserves the antioxidant activity associated with this lipoprotein fraction. PMID- 14519783 TI - Dietary amino acids promote pancreatic protease synthesis at the translation stage in rats. AB - In some tissues, amino acids (AA) stimulate translation initiation via interactions between eukaryote initiation factor (eIF) 4E-binding protein 1 (4E BP1), eIF4E and eIF4G. Dietary AA have been shown to induce pancreatic proteases independently of cholecystokinin in rats, the mechanism of which has not yet been clarified. In the present study, we examined the mechanism in rats for protease induction by dietary AA and determined the involvement of translation initiation. Male Wistar/ST rats were fed a 20 or 60% casein or AA mixture diet for 7 d and were intravenously injected with [35S] methionine (Met) 30 min before killing on d 7 (expt. 1). In expt. 2, rats were fed a 20 or 60% AA diet for 7 d and after food deprivation and refeeding with the respective diet on d 7 were killed at 0, 1 or 3 h. We measured mRNA and [35S] Met incorporation into chymotrypsinogen, phosphorylation status of 4E-BP1 and the association of eIF4E with 4E-BP1 or eIF4G. In expt. 1, chymotrypsin activity and synthesis were higher in both of the 60% diet groups than in the 20% diet groups, but the mRNA level and 4E-BP1 status did not differ. In expt. 2, chymotrypsin activity increased in the 60% AA diet group in a time-dependent manner. The translation initiation activity via the mTOR pathway indicated an increase similar to chymotrypsin activity. There were no differences in chymotrypsin mRNA level at any point. These results indicate that dietary AA induce chymotrypsin synthesis by promoting translation, and transient activation of translation initiation via mTOR may be associated with this induction. PMID- 14519784 TI - Zinc modulates PPARgamma signaling and activation of porcine endothelial cells. AB - Dietary zinc has potent antioxidant and anti-inflammatory properties and is a critical component of peroxisome proliferator-activated receptor (PPAR) gene expression and regulation. To assess the protective mechanisms of PPARgamma in endothelial cell dysfunction and the role of zinc in the modulation of PPARgamma signaling, cultured porcine pulmonary artery endothelial cells were exposed to the membrane-permeable zinc chelator N,N,N'N'-tetrakis (2-pyridylmethyl)-ethylene diamine (TPEN), thiazolidinedione (TZD; PPARgamma agonist) or bisphenol A diglycidyl ether (BADGE; PPARgamma antagonist). Subsequently, endothelial cells were activated by treatment with linoleic acid (90 micro mol/L) for 6 h. Zinc chelation by TPEN increased the DNA binding activity of nuclear factor (NF) kappaB and activator protein (AP)-1, decreased PPARgamma expression and activation as well as up-regulated interleukin (IL)-6 expression and production. These effects were fully reversed by zinc supplementation. In addition, exposure to TZD down-regulated linoleic acid-induced DNA binding activity of NF-kappaB and AP-1, whereas BADGE further induced activation of these oxidative stress sensitive transcription factors. Most importantly, the TZD-mediated down regulation of NF-kappaB and AP-1 and reduced inflammatory response were impaired during zinc chelation. These data suggest that zinc plays a critical role in PPARgamma signaling in linoleic acid-induced endothelial cell activation and indicate that PPARgamma signaling is impaired during zinc deficiency. PMID- 14519785 TI - Glutamine prevents cytokine-induced apoptosis in human colonic epithelial cells. AB - Epithelial cell apoptosis is an important regulator of normal gut mucosal turnover; however, excessive apoptosis may inhibit mucosal restitution during pathophysiologic states. Apoptosis is induced by oxidative stress and cytokines, but regulation by specific nutrients has been infrequently studied under these conditions. Glutamine (Gln) is an important metabolic fuel for intestinal epithelial cells and a precursor to the antioxidant glutathione (GSH), which has antiapoptotic effects. In cultured intestinal epithelial cells, Gln depletion increases oxidant-induced apoptosis. This study examined whether Gln protects against apoptosis induced by the cytokine tumor necrosis factor-alpha-related apoptosis-inducing ligand (TRAIL) in the human colon carcinoma cell line, HT-29. TRAIL-induced apoptosis in HT-29 cells was characterized by an increase in the percentage of cells in the sub-G1 fraction by flow cytometry, nuclear condensation and the activation of caspase-8 and caspase-3. TRAIL-induced apoptosis was completely prevented by Gln, but not inhibited by other amino acids, including the GSH constituents, glutamate, cysteine and glycine. Similar antiapoptotic effects of Gln occurred when apoptosis was induced by a combination of tumor necrosis factor-alpha and interferon-gamma. Cellular GSH was oxidized during TRAIL-induced apoptosis. This effect was completely blocked by Gln, however, inhibition of GSH synthesis with buthionine sulfoximine did not alter Gln antiapoptotic effects. Furthermore, glutamate prevented GSH oxidation in response to TRAIL but did not protect against TRAIL-induced apoptosis. These results show that Gln specifically protects intestinal epithelial cells against cytokine-induced apoptosis, and that this occurs by a mechanism that is distinct from the protection against oxidative stress mediated by cellular GSH. PMID- 14519786 TI - Dietary and hormonal stimulation of rat exocrine pancreatic function regulates CRHSP-28 phosphorylation in vivo. AB - Dietary regulation of digestive enzyme secretion from the pancreas is essential for the breakdown of macronutrients in the gastrointestinal tract. Ca(2+) responsive heat stable protein (CRHSP)-28 is a regulatory protein that modulates the exocytosis of digestive enzymes from pancreatic acinar cells. In the present study, isoelectric focusing and immunoblotting were used to characterize CRHSP-28 phosphorylation in isolated rat acinar cells and also after hormonal and dietary stimulation of rat pancreas in vivo. CRHSP-28 was highly phosphorylated in isolated acini after stimulation with a physiologic range of concentrations of cholecystokinin-octapeptide (CCK-8). Activation of the high affinity state of the CCK-A receptor with the synthetic peptide JMV-180 confirmed the physiologic relevance of the response. CRHSP-28 phosphorylation was contingent on elevated cellular Ca2+ because it was maximally stimulated by Ca2+ ionophore, but unchanged after protein kinase C, cAMP or cyclic guanosine monophosphate activation. Intravenous infusion of rats with a secretory concentration of the CCK analog, caerulein, stimulated CRHSP-28 phosphorylation by 100% over control (P < 0.01) within 15 min of dosing. Moreover, CRHSP-28 phosphorylation was stimulated by 150% over control (P < 0.05) immediately after consumption of a semipurified AIN-93 diet. These data demonstrate that CRHSP-28 phosphorylation occurs in vivo and can be used as a functional indicator of nutrient-driven acinar cell activation. PMID- 14519787 TI - Transgenic chickens expressing beta-galactosidase hydrolyze lactose in the intestine. AB - Chickens do not possess the necessary enzymes to efficiently hydrolyze lactose into glucose and galactose. The bacterial enzyme beta-galactosidase can convert lactose into glucose and galactose. Transgenic chickens that carry the E. coli lacZ gene and express beta-galactosidase could potentially utilize lactose as an energy source. The objective of this study was to determine the ability of the transgenic chicken small intestinal mucosa to hydrolyze lactose into glucose and galactose. Lactase activity was examined in the intestinal muscosa from wild-type chickens and two lines of chickens that carry the lacZ gene and express beta galactosidase. Lactase activity was significantly higher in both transgenic lines compared with wild-type birds (P < 0.05). The presence of the beta-galactosidase enzyme was revealed by X-gal staining in the intestine of transgenic chickens, whereas it was not present in the wild-type chickens. Overall, it appears that inserting the lacZ gene, which encodes beta-galactosidase, has resulted in a chicken that can utilize lactose as an energy source. This study demonstrates that transgenic technology can be used to modify nutrient utilization in domestic poultry. PMID- 14519788 TI - A pilot study to investigate effects of inulin on Caco-2 cells through in vitro metabolic fingerprinting. AB - Metabolic fingerprints are novel measurement tools to evaluate the biochemical status of a living organism by using 1H NMR and multivariate data analysis (MVDA). In this way, a quick evaluation of changes in health or diseased state can be made, reflected in alterations of metabolic patterns. Normally, metabolic fingerprinting is based on in vivo studies. These studies often represent a labor intensive and expensive manner of investigation. In vitro studies are not hampered by these disadvantages, thus constituting an interesting alternative. In this research, results are presented of a pilot experiment in which metabolic fingerprinting was combined with an in vitro model. For this purpose, differentiated Caco-2 cells were exposed to inulin and its fermentative metabolites, both dissolved in culture medium. Cells were incubated for 0 or 48 h. Cell fractions were analyzed by NMR, then subsequently with MVDA. Differences in treatment provided detectable variations in the time of metabolic patterns of cell contents. Results indicated that glucose metabolism linked to glutamate was of major importance in the effects of inulin and its metabolites on Caco-2 cells under the conditions of our study. Metabolic fingerprinting in combination with an in vitro model appears to be a feasible technique with which to visualize metabolic patterns of cell contents and provides an efficient place for the generation of hypotheses about the metabolic pathways involved. In vitro metabolic fingerprinting may be of great benefit in the future for a better understanding of the relationship between nutrition and health. PMID- 14519789 TI - Fetal and early postnatal protein malnutrition cause long-term changes in rat liver and muscle mitochondria. AB - Epidemiologic data suggest a strong association between low birth weight and increased risk of metabolic syndrome in adult life. However, the underlying mechanisms are largely unknown. To test the hypothesis that mitochondrial changes may serve as a link between poor nutrition in early life and insulin resistance in later life, we investigated the effect of protein malnutrition during gestation and lactation on mitochondria of the liver and skeletal muscle. Female offspring of Sprague-Dawley rats fed a low protein diet (casein, 80 g/kg) were randomly divided into two groups and weaned onto either the low protein diet or a control diet (casein, 180 g/kg). As a control group, offspring of rats fed the control diet were weaned onto the control diet. The rats in each group were randomly divided into four groups that were killed at 5, 10, 15 and 20 wk of age. Both mitochondrial DNA content and the expression of mitochondrial DNA-encoded genes in liver and muscle were measured. Mitochondrial transcription factor A and antioxidant enzyme activities were also determined. The mitochondrial DNA content of the liver and skeletal muscle were reduced in fetal and early postnatal malnourished rats even when proper nutrition was supplied after weaning. These changes were accompanied by a decrease in mitochondrial DNA-encoded gene expression; however, they were not dependent on mitochondrial transcriptional factor A. Our findings indicate that poor nutrition in early life causes long lasting changes in mitochondria that may contribute to the development of insulin resistance in later life. PMID- 14519790 TI - Selenoprotein-deficient transgenic mice exhibit enhanced exercise-induced muscle growth. AB - Dietary intake of selenium has been implicated in a wide range of health issues, including aging, heart disease and cancer. Selenium deficiency, which can reduce selenoprotein levels, has been associated with several striated muscle pathologies. To investigate the role of selenoproteins in skeletal muscle biology, we used a transgenic mouse (referred to as i6A-) that has reduced levels of selenoproteins due to the introduction and expression of a dominantly acting mutant form of selenocysteine transfer RNA (tRNA[Ser]Sec). As a consequence, each organ contains reduced levels of most selenoproteins, yet these mice are normal with regard to fertility, overall health, behavior and blood chemistries. In the present study, although skeletal muscles from i6A- mice were phenotypically indistinguishable from those of wild-type mice, plantaris muscles were approximately 50% heavier after synergist ablation, a model of exercise overload. Like muscle in wild-type mice, the enhanced growth in the i6A- mice was completely blocked by inhibition of the mammalian target of rapamycin (mTOR) pathway. Muscles of transgenic mice exhibited increased site-specific phosphorylation on both Akt and p70 ribosomal S6 kinase (p70S6k) (P < 0.05) before ablation, perhaps accounting for the enhanced response to synergist ablation. Thus, a single genetic alteration resulted in enhanced skeletal muscle adaptation after exercise, and this is likely through subtle changes in the resting phosphorylation state of growth-related kinases. PMID- 14519791 TI - Diet-induced milk fat depression in dairy cows results in increased trans-10, cis 12 CLA in milk fat and coordinate suppression of mRNA abundance for mammary enzymes involved in milk fat synthesis. AB - Milk composition can be altered by diet, and one example is milk fat depression (MFD) in dairy cows. The biohydrogenation theory of MFD has implicated unique fatty acids formed by altered rumen biohydrogenation of PUFA; one example is trans-10, cis-12 conjugated linoleic acid (CLA). In the present study, we induced MFD with a high concentrate/low forage (HC/LF) diet and examined milk composition, milk fatty acid changes and mammary lipogenic mRNA abundance to determine the mechanism involved. The HC/LF diet reduced milk fat percentage by 25% and yield by 27% with no effect on dietary intake, milk production, protein or lactose. Milk fatty acids synthesized de novo in the mammary gland and fatty acids taken up from circulation were reduced to a similar extent (molar basis). MFD was also characterized by the appearance of trans-10, cis-12 CLA in the milk fat. We analyzed mammary mRNA abundance for lipogenic genes and detected reductions for acetyl CoA carboxylase (ACC), fatty acid synthase (FAS), fatty acyl CoA ligase, glycerol phosphate acyltransferase (GPAT) and acylglycerol phosphate acyltransferase (AGPAT). There was no effect on the milk protein gene, kappa-casein. The reductions in mRNA were also correlated with the appearance of trans-10, cis-12 CLA in the milk fat for ACC, FAS, lipoprotein lipase and GPAT. This study demonstrates that diet-induced MFD involves coordinated effects on mRNA for mammary lipid synthesis pathways, and provides support for a mechanism involving alterations in transcriptional activation of these genes. PMID- 14519792 TI - Bakery products enriched with phytosterol esters, alpha-tocopherol and beta carotene decrease plasma LDL-cholesterol and maintain plasma beta-carotene concentrations in normocholesterolemic men and women. AB - The hypocholesterolemic effects of phytosterols have not been evaluated in bakery products, and the addition of liposoluble antioxidants to the carrier has never been tested. We investigated the effects of consuming croissants and magdalenas (Spanish muffins) enriched with sterol esters, alpha-tocopherol and beta-carotene on plasma lipid and fat-soluble antioxidant concentrations in normocholesterolemic, habitual consumers of bakery products following their usual diet and lifestyle. Using a randomized, double-blind, placebo-controlled design, the control (C) group (n = 29) received two pieces daily (standard croissant and muffin) and the sterol ester (SE) group (n = 28), the same products with sterol esters added (3.2 g/d) for 8 wk. Total and LDL cholesterol (LDL-C) decreased in the SE group by 0.24 mmol/L (P < 0.01) and 0.26 mmol/L (P < 0.005), respectively, whereas these variables did not change in the control group. The total difference in total and LDL-C changes between groups was 0.38 mmol/L (8.9%) and 0.36 mmol/L (14.7%), respectively (P < 0.001). Within-group changes in HDL cholesterol, triacylglycerol or lipoprotein(a) concentrations did not differ. Similarly, within-group changes over time in plasma tocopherol and carotenoid concentrations did not differ between groups. Our findings suggest that bakery products are excellent carriers for phytosterols, and their consumption is associated with a decrease in total and LDL-C concentrations, with no changes in alpha-tocopherol and beta-carotene. The ability of bakery products to include sufficient quantities of beta-carotene to compensate for a potential deficiency, and the fact that their efficacy was not associated with the time of day at which they were consumed, are interesting findings. PMID- 14519793 TI - Rapid gut transit time and slow fecal isoflavone disappearance phenotype are associated with greater genistein bioavailability in women. AB - The bioavailability of soybean isoflavones varies widely among individuals due to many factors, including activities of gut microflora. To characterize factors that affect fecal isoflavone disappearance phenotype and isoflavone bioavailability in women, 35 Asian and 33 Caucasian women, 18-43 y of age, provided fecal samples for anaerobic incubation with isoflavones in vitro at two times 5 mo apart (Phases I and II). Diet, physical activity and health history were investigated at these times. A single dose of soymilk powder [1.2 mg (4.57 micromol) total isoflavone/kg body] was given to all subjects with breakfast in phase II. Daidzein and genistein from fecal incubations, urine and fecal samples were measured by reverse-phase HPLC. Three significantly different daidzein and two genistein disappearance phenotypes were identified from fecal isoflavone incubations. More Asians than Caucasians were identified within the high daidzein disappearance phenotype. Caucasians and Asians differed significantly in daily intake of red meat (0.3 +/- 0.1 vs. 1.0 +/- 0.1 servings/d), dairy foods (2.9 +/- 0.2 vs. 1.2 +/- 0.2 servings/d) and insoluble dietary fiber (3.6 +/- 0.3 vs. 1.4 +/- 0.3 g). BMI, maximal oxygen uptake (VO(2 max)) and physical activity level were significantly greater in Caucasians than in Asians. Asian subjects of the low genistein disappearance phenotype had more rapid gut transit time (GTT) and greater isoflavone bioavailability as reflected in urinary genistein excretion than did Asians of the high genistein disappearance phenotype (GTT, 40 +/- 8 vs. 63 +/- 5 h; 11.0 +/- 2.7 vs. 4.0 +/- 1.7% of ingested genistein excreted in urine). Caucasians of both genistein disappearance phenotypes had longer GTT than did Asian subjects (84 +/- 5 vs. 56 +/- 6 h) and resembled Asians of the high genistein disappearance phenotype in genistein bioavailability. Relatively rapid GTT coupled with a low fecal isoflavone disappearance phenotype as occurred in Asian but not Caucasian subjects produced greater genistein bioavailability, as reflected in urinary genistein excretion. PMID- 14519794 TI - Dietary modification and moderate antioxidant supplementation differentially affect serum carotenoids, antioxidant levels and markers of oxidative stress in older humans. AB - Oxidative stress is thought to be a factor in the progression of chronic diseases, and even the aging process itself. We examined the effect of two moderate vitamin, mineral and phytochemical supplements and a dietary intervention on markers of oxidative stress and antioxidant status in older humans. Subjects (n = 60, 65-85 y old) were enrolled in a 5-wk, double-blind, placebo-controlled study. The subjects were randomly assigned to a group with one of four interventions: antioxidant supplement capsule, antioxidant supplement tablet, high carotenoid diet or placebo. Postintervention serum levels of some of the supplemented carotenoids and alpha-tocopherol were higher in all treated groups compared with the placebo group and with preintervention levels within groups. Markers of oxidative stress or antioxidant capacity were not significantly different from preintervention levels, although the diet and supplement capsule groups had lower levels of some of those markers than the placebo group postintervention. The results suggest that both moderate antioxidant supplementation and a diet high in carotenoids elevate serum carotenoids and antioxidant levels in an older adult population, although with different specific effects. PMID- 14519795 TI - High carbohydrate and high monounsaturated fatty acid diets similarly affect LDL electrophoretic characteristics in men who are losing weight. AB - We compared the effects of ad libitum consumption of a defined high complex carbohydrate (CHO) diet (% of energy: CHO, 58.3; fat, 25.8) vs. a defined high monounsaturated fatty acid (MUFA) diet (% of energy: CHO, 44.7; fat, 40.1; MUFA, 22.5) on LDL electrophoretic characteristics. Healthy men [n = 65; age, 37.5 +/- 11.2 (mean +/- SD) y; BMI, 29.2 +/- 4.9 kg/m2] were randomly assigned to one of the two diets that they consumed for 6-7 wk. The high CHO diet significantly reduced body weight (-2%). The diet-induced reduction in plasma LDL cholesterol (C) levels in the high-CHO diet group was due mainly to concurrent reductions in the cholesterol content of small (<25.5 nm, P < 0.01) and medium-sized LDL (25.5 26.0 nm, P = 0.01). The high MUFA diet also reduced body weight, and LDL-C and LDL-apolipoprotein (apo)B levels, which were comparable to those in the high CHO group. The cholesterol levels of small LDL particles tended to be reduced (P = 0.24) in the high MUFA group (-12%), similar to changes in the high CHO group. These results suggest that, when associated with weight loss, ad libitum consumption of high CHO and high MUFA diets may be considered to be equally beneficial for the management of LDL-related atherogenic dyslipidemia. However, the high MUFA diet more favorably affected triglyceride levels, suggesting that it may be preferable to a high CHO diet in cardiovascular disease prevention. PMID- 14519796 TI - The DASH diet and sodium reduction improve markers of bone turnover and calcium metabolism in adults. AB - Dietary strategies to prevent and treat osteoporosis focus on increased intake of calcium and vitamin D. Modification of whole dietary patterns and sodium reduction may also be effective. We examined the effects of two dietary patterns and three sodium levels on bone and calcium metabolism in a randomized feeding study. A total of 186 adults, aged 23-76 y, participated. After a 2-wk run-in period, participants were assigned randomly to diets containing three levels of sodium (50, 100 and 150 mmol/d) to be consumed for 30 d in random order. Serum osteocalcin (OC), C-terminal telopeptide of type I collagen (CTX), fasting serum parathyroid hormone (PTH), urinary sodium, potassium, calcium and cAMP were measured at baseline and at the end of each sodium period. The Dietary Approaches to Stop Hypertension (DASH) diet reduced serum OC by 8-11% and CTX by 16-18% (both P < 0.001). Urinary calcium excretion did not differ between subjects that consumed the DASH and control diets. Reducing sodium from the high to the low level significantly decreased serum OC 0.6 microg/L in subjects that consumed the DASH diet, fasting serum PTH 2.66 ng/L in control subjects and urinary calcium 0.5 mmol/24 h in both groups. There were no consistent effects of the diets or sodium levels on urinary cAMP. In conclusion, the DASH diet significantly reduced bone turnover, which if sustained may improve bone mineral status. A reduced sodium intake reduced calcium excretion in both diet groups and serum OC in the DASH group. The DASH diet and reduced sodium intake may have complementary, beneficial effects on bone health. PMID- 14519797 TI - Supplementation of diets with alpha-tocopherol reduces serum concentrations of gamma- and delta-tocopherol in humans. AB - Despite promising evidence from in vitro experiments and observational studies, supplementation of diets with alpha-tocopherol has not reduced the risk of cardiovascular disease and cancer in most large-scale clinical trials. One plausible explanation is that the potential health benefits of alpha-tocopherol supplements are offset by deleterious changes in the bioavailability and/or bioactivity of other nutrients. We studied the effects of supplementing diets with RRR-alpha-tocopheryl acetate (400 IU/d) on serum concentrations of gamma- and delta-tocopherol in a randomized, placebo-controlled trial in 184 adult nonsmokers. Outcomes were changes in serum concentrations of gamma- and delta tocopherol from baseline to the end of the 2-mo experimental period. Compared with placebo, supplementation with alpha-tocopherol reduced serum gamma tocopherol concentrations by a median change of 58% [95% CI = (51%, 66%), P < 0.0001], and reduced the number of individuals with detectable delta-tocopherol concentrations (P < 0.0001). Consistent with trial results were the results from baseline cross-sectional analyses, in which prior vitamin E supplement users had significantly lower serum gamma-tocopherol than nonusers. In view of the potential benefits of gamma- and delta-tocopherol, the efficacy of alpha tocopherol supplementation may be reduced due to decreases in serum gamma- and delta-tocopherol levels. Additional research is clearly warranted. PMID- 14519798 TI - Fermentable and nonfermentable fiber supplements did not alter hunger, satiety or body weight in a pilot study of men and women consuming self-selected diets. AB - Little is known about the relative effects of fermentable fiber (FF) vs. nonfermentable fiber (NFF) on energy regulation in humans. We compared 27 +/- 0.6 g/d supplements of FF (pectin, beta-glucan) and NFF (methylcellulose) for their ability to decrease ad libitum energy intake (EI) and hunger, increase satiety and cause spontaneous body weight and fat losses. Men and women (n = 11) aged 23 46 y, BMI 20.0-34.4 kg/m2, consumed first NFF and then FF for 3 wk each, with a 4 wk washout period between phases. Daily satiety assessed with analog scales was higher with NFF than FF (60.7 +/- 1.0 vs. 57.7 +/- 0.8 mm, P = 0.01). However, there were no differences in reported EI (NFF < FF by 7%, P = 0.31, NFF < baseline by 9.5%, P = 0.11), body weight (NFF 0.13 kg, P = 0.73; FF 0.13 kg, P = 0.60) or fat percentage (NFF -0.3%, P = 0.56; FF -0.1%, P = 0.66) within either phase. In contrast to findings in animals, NFF was more, rather than less satiating than FF, and use of neither NFF nor FF preparations was associated with body weight or fat loss. These pilot results suggest no role for short-term use of FF and NFF supplements in promoting weight loss in humans consuming a diet ad libitum. PMID- 14519799 TI - Fecal acetate is inversely related to acetate absorption from the human rectum and distal colon. AB - In humans, colonic bacteria ferment unabsorbed carbohydrates, producing the SCFA acetic, propionic and n-butyric acids. To test for interactions among the SCFA that may affect their absorption, healthy subjects (n = 10) were given 300-mL rectal infusions containing acetate (60 mmol/L), propionate (20 mmol/L) and butyrate (20 mmol/L), alone or in combinations of two or three. The solutions were retained for 30 min, and then subjects voided a sample for SCFA measurement. To examine the relationship between absorption and fecal SCFA concentrations, a fecal sample was collected at the end of the study. The mean percentage of butyrate absorption (30.2 +/- 4.6%) exceeded that of acetate (24.1 +/- 3.7%) (P < 0.05). Absorption tended to be less (P = 0.12) when a SCFA was infused alone (26.7 +/- 4.0%) than when all three were infused (32.0 +/- 5.7%). Bicarbonate concentration was higher after butyrate-containing infusions than after saline. The fecal molar acetate percentage was inversely correlated with the percentage of acetate absorption from the infusion of three SCFA (r = -0.834, P < 0.005). We conclude that there was no combination effect on SCFA absorption, and the chain length effect suggests passive diffusion as a likely mechanism of absorption. Furthermore, fecal acetate may reflect absorption, rather than production of colonic acetate. PMID- 14519800 TI - Cocoa powder increases postprandial insulinemia in lean young adults. AB - We hypothesized that chocolate products elicit higher insulin responses than matched products with alternate flavoring. To test this, we used a within subject, repeated-measures comparison of six pairs of foods, one flavored with chocolate (cocoa powder) and the other not. Healthy subjects (n = 10, 4 men, 6 women) tested each pair of foods. Postprandial glucose and insulin levels were determined at intervals over 2 h using standardized glycemic index (GI) methodology. The product categories were chocolate bars, cakes, breakfast cereals, ice creams, flavored milks and puddings. Although the GI did not differ within each pair, the insulin index (II) of the chocolate product was always higher, by a mean of 28%, than the alternate flavored product (P < 0.001). The greatest difference occurred within the flavored milk category in which the chocolate version elicited 45% greater insulinemia than the strawberry flavored milk (P = 0.021). Macronutrient composition (fat, protein, sugar, fiber or energy density) accounted for nearly all of the variation in GI among the foods, but did not explain differences in insulinemia. The presence of cocoa powder in foods leads to greater postprandial insulin secretion than alternate flavorings. Specific insulinogenic amino acids or greater cephalic phase insulin release may explain the findings. PMID- 14519801 TI - Replacement-fed infants born to HIV-infected mothers in India have a high early postpartum rate of hospitalization. AB - Access to safe breast-feeding alternatives for HIV-infected mothers and their infants in many settings is limited. We compared the rates of early postpartum hospitalization of infants born to HIV-infected mothers using different infant feeding practices in a large government hospital in Pune, India. From March 1, 2000 to November 30, 2001, infants born to HIV-infected mothers were followed in a postpartum clinic. All mothers had received a standard short course of antenatal zidovudine. Infant-feeding practices were assessed within 3 d of delivery, prior to postpartum hospital discharge. Sixty-two of 148 mothers (42%) were breast-feeding their infants. Eighty-six of the mothers (58%) were providing replacement feeding, primarily diluted cow, goat or buffalo milk (top feeding). Twenty-one of the 148 participating infants (14.2%) born during the study period required hospitalization within the 1st 6 mo of life and 6 infants required repeat hospitalization. All hospitalized infants were receiving replacement feeding with a rate of 0.093 hospitalizations per 100 person-days (95% CI, 0.062 to 0.136). The reasons for hospitalization included acute gastroenteritis (48.1%), pneumonia (18.5%), septicemia (11.1%) and jaundice (11.1%). A high risk for early postpartum hospitalization was seen in replacement-fed infants born to HIV-infected mothers in Pune, India. In settings such as India, where access to safe replacement feeding is limited, interventions making exclusive breast feeding safer for HIV-infected mothers and infants are needed. Such interventions would be valuable additions to the very effective national prevention programs that currently rely on the provision of short-course zidovudine and nevirapine. PMID- 14519802 TI - Iron bioavailability in corn-masa tortillas is improved by the addition of disodium EDTA. AB - Corn-masa flour flat bread tortillas are the main staple of Mexican and Central American populations. Due to high concentrations of inhibitors of iron absorption, the bioavailability from this matrix is unknown. We wanted to determine the most suitable fortificant that would efficaciously improve iron bioavailability. In tortillas prepared with commercial precooked, lime-treated, corn-masa flour, we examined the in vitro solubility of the following forms of iron: native iron with and without Na2EDTA, elemental reduced iron plus Na2EDTA, ferrous fumarate with and without Na2EDTA, bisglycine iron, ferrous sulfate and NaFeEDTA. We also examined the in vivo bioavailability in humans with double radioiron erythrocyte incorporation of ferrous fumarate with and without Na2EDTA, bisglycine iron, NaFeEDTA and native iron plus Na2EDTA, beans and rice. In vitro, solubility ranged from 1% in iron forms without Na2EDTA to 19.4% for NaFeEDTA. Forms of iron with Na2EDTA had intermediate values. In vivo radioiron studies showed that iron forms without Na2EDTA also had low bioavailability (< or =1%). NaFeEDTA had the highest bioavailability (5.3%). The bioavailability of all iron forms improved significantly when tested with Na2EDTA (<0.05). Adding Na2EDTA to ferrous fumarate increased bioavailability from 0.87% to 2.9% (P < 0.001). We conclude that NaFeEDTA is the form of iron best absorbed, but alternatively, ferrous fumarate plus Na2EDTA comprises a feasible option as a fortificant. PMID- 14519803 TI - Subclinical prenatal iodine deficiency negatively affects infant development in Northern China. AB - Although mild-to-moderate levels of iodine deficiency (ID) have been associated with poor cognitive outcomes in children, little is known about subclinical prenatal ID and infant development. In this study, the association between elevated cord blood thyroid stimulating hormone (TSH, thyrotropin) and infant development was examined in Northern China. Three groups of infants with elevated cord blood TSH were compared with infants with normal TSH levels on an information processing task at 7 mo, and in cognitive and motor developmental assessments at 13 mo. Infants with elevated TSH had poorer information processing skills and lower scores on the cognitive development index. There were no differences in motor abilities. Relationships between socioenvironmental factors and iodine status were assessed. Infants from more rural settings and those whose mothers had completed fewer years of schooling and had lower paying occupations had higher cord blood TSH levels. A regression analysis indicated that maternal education was predictive of cognitive performance among infants with elevated TSH but not control infants. The findings suggest that subclinical prenatal ID has negative effects on infant development and that, in some instances, maternal education may ameliorate these effects. PMID- 14519804 TI - Consumption of folic acid-fortified bread improves folate status in women of reproductive age in Chile. AB - Since January 2000 the Chilean Ministry of Health has required the fortification of wheat flour with folic acid (FA) at a concentration of 2.2 mg FA/kg in order to reduce the risk of neural tube defects (NTD) in newborns. This policy was expected to result in a mean additional intake of approximately 400 microg FA/d. We assessed the effectiveness of the FA flour fortification program on bread folate content and on blood folate concentration in women of childbearing age in Santiago, Chile. The prefortification folate status of 751 healthy women of reproductive age was assessed. The folate content of 100 bread samples bought at retail bakeries was measured, average wheat flour consumption was estimated and postfortification FA dietary intake was calculated. The effect of flour fortification on blood folate concentration in this group of women (n = 605) was evaluated in a follow-up study. Blood folate concentrations of the 605 women in the follow-up group increased (P < 0.0001) following fortification. Before fortification the mean serum and red blood cell folate concentrations were 9.7 +/ 4.3 and 290 +/- 102 nmol/L, respectively, compared with 37.2 +/- 9.5 and 707 +/- 179 nmol/L postfortification, respectively. The mean FA content of bread was 2020 +/- 940 micro g/kg. The median FA intake of the group evaluated postfortification was 427 microg/d (95% CI 409-445) based on an estimated intake of 219 g/d (95% CI 201-229) of wheat flour, mainly as bread. Fortification of wheat flour substantially improved folate status in a population of women of reproductive age in Chile. The effect of the FA fortification program on the occurrence of NTD is currently being assessed. PMID- 14519805 TI - Fish oil supplementation of rats during pregnancy reduces adult disease risks in their offspring. AB - Metabolic programming in utero due to maternal undernutrition is considered to increase the risk of adult diseases in offspring. It is therefore of relevance to investigate how dietary supplementation of specific nutrients can ameliorate the negative effects of maternal malnutrition. We examined the effects of supplementing fish oil or folic acid, both of which are conventional supplements in maternal intervention, on risk factors in the offspring as adults. Pregnant female rats from 4 groups (n = 6/group) were fed casein diets with 18 g/100 g protein (control diet), 12 g/100 g protein supplemented with 8 mg folic acid/kg diet (0.08 mg/kg diet) (FAS), 12 g/100 g protein without folic acid (FAD) or 12 g/100 g protein supplemented with 7 g/100 g fish oil (FOIL). Pups were weaned to a standard laboratory diet with 18 g/100 g protein. Serum glucose, insulin and cholesterol and plasma homocysteine levels were measured in the offspring at 6 and 11 mo of age. Serum glucose in 11-mo-old male and female pups was greater (P < 0.05) in both the FAS (males 2.46 +/- 0.51, females 2.49 +/- 0.29 mmol/L) and FAD groups (2.48 +/- 0.28 and 2.67 +/- 0.41 mmol/L) than in controls (2.03 +/- 0.15 and 2.02 +/- 0.18 mmol/L). Serum insulin concentrations were higher (P < 0.05) in the FAD group (males 1476 +/- 317, females 1441 +/- 220 pmol/L) but were lower in males from the FAS group (483 +/- 165 pmol/L) compared with controls (males 917 +/- 373, females 981 +/- 264 pmol/L). Glucose and insulin concentrations did not differ between the control and FOIL groups. Plasma homocysteine levels were lower (P < 0.05) only in 11-mo-old folate-deficient males; none of the other groups differed from the controls. Maternal supplementation of fish oil to a diet containing marginal protein was beneficial in maintaining circulating glucose, insulin, cholesterol and homocysteine levels in the offspring as adults. PMID- 14519806 TI - Coenzyme Q intake elevates the mitochondrial and tissue levels of Coenzyme Q and alpha-tocopherol in young mice. AB - The main objective of this study was to resolve the issue of whether the amounts of Coenzyme Q (CoQ), which is endogenously synthesized in cells, can be elevated in tissues and mitochondria of young mice by dietary supplementation with CoQ10. The prevalent view is that the uptake of exogenous CoQ by tissues other than plasma and liver either does not occur or is quite minimal. Mice, 6 mo of age, were fed 0, 148 or 654 mg CoQ10/(kg body x d) in their diets for 11 wk. CoQ10 intake enhanced both CoQ9 and CoQ10 homologues in the plasma, and in homogenates and mitochondria of liver, heart and skeletal muscle. CoQ was elevated in brain mitochondria, but not in the brain homogenate. The uptake of exogenous CoQ was higher in mitochondria of heart and skeletal muscle than those in liver. CoQ10 administration also elevated the alpha-tocopherol concentration in tissue homogenates and their mitochondria, thereby providing an in vivo indication of the "sparing" effect of CoQ on alpha-tocopherol. Results of this study demonstrate that, contrary to the historical view, both total and mitochondrial CoQ concentrations in the heart and skeletal muscle and in the mitochondria of brain of young mice can be augmented by dietary supplementation. Furthermore, CoQ intake enhances the antioxidative potential of tissues by elevating the endogenous amounts of alpha-tocopherol. PMID- 14519807 TI - Dietary conjugated linoleic acids as free fatty acids and triacylglycerols similarly affect body composition and energy balance in mice. AB - The objective of this study was to compare the effects of conjugated linoleic acid (CLA) as triacylglycerols (TAG) or free fatty acids (FFA) on body composition and energy balance in mice. We fed four groups of 5-wk-old Balb-C mice (n = 9) semipurified diets containing either CLA (0.5 g CLA/100 g of diet) or high oleic sunflower oil (HOSF) in the form of FFA or TAG for 42 d. Body composition was determined and the energy in the carcasses, excreta and food was measured in a bomb calorimeter. The amount of body fat was 4.72 +/- 0.95 g (17.9 +/- 2.8%) in the HOSF-FFA group, 2.36 +/- 0.29 g (9.4 +/- 1.0%) in the CLA-FFA mice (mean +/- SD, P < 0.05), 4.76 +/- 0.74 g (18.2 +/- 2.2%) in the HOSF-TAG group and 2.32 +/- 0.38 g (9.3 +/- 1.1%) in the CLA-TAG mice (P < 0.05). The percentage of energy intake that was stored in the body was 3.5 +/- 1.2% in the HOSF-FFA group, 0.6 +/- 0.3% in the CLA-FFA group (P < 0.05), 3.5 +/- 1.1% in the HOSF-TAG group and 0.5 +/- 0.4 in the CLA-TAG mice (P < 0.05). Conversely, the percentage of energy intake that was expended as heat was 89.4 +/- 1.2% in the HOSF-FFA group, 92.4 +/- 0.8% in the CLA-FFA mice (P < 0.05), 89.47 +/- 1.23% in the HOSF-TAG group and 92.2 +/- 0.4% in the CLA-TAG group (P < 0.05). Thus, CLA in the form of FFA or TAG had similar effects on body composition and energy balance. PMID- 14519808 TI - Megasphaera elsdenii JCM1772T normalizes hyperlactate production in the large intestine of fructooligosaccharide-fed rats by stimulating butyrate production. AB - Hyperlactate production is related to disorders of the large intestine such as inflammatory bowel diseases. Lactate, an intermediate in hindgut fermentation, is metabolized to SCFA. Megasphaera elsdenii can convert lactate to butyrate, a physiologically important organic acid for the hindgut mucosa. This experiment was conducted to determine whether M. elsdenii normalizes hyperlactate production and stimulates butyrate production in the rat large intestine. Specific pathogen free Sprague-Dawley male rats (n = 12) were fed a fructooligosaccharide (FOS) supplemented (100 g/kg), semipurified diet to induce lactate production. Lactate excretion in all rats was >30 mmol/kg fresh feces on d 2 of FOS-feeding. The rats were divided into two groups on the morning of d 4. One group (n = 5) was dosed orally with M. elsdenii JCM1772T (1.3 x 10(13) cells) for 3 d. The other group was treated with a vehicle solution. Fecal lactate was significantly lower in rats administered M. elsdenii than in controls. An increase in fecal butyrate compensated for the decrease in lactate. The number of cecal epithelial cells was greater in rats administered M. elsdenii than in controls. M. elsdenii has the potential to normalize hyperlactate accumulation in the large intestine, and lactate-utilizing butyrate producers may be useful probiotics when hyperlactate fermentation in the large intestine is a problem. PMID- 14519809 TI - Assessment of vitamin B-6 estimated average requirement and recommended dietary allowance for adolescents aged 13-15 years using vitamin B-6 intake, nutritional status and anthropometry. AB - Because of limited available information, the Estimated Average Requirement (EAR) and Recommended Dietary Allowance (RDA) of vitamin B-6 for adolescents were recently estimated by extrapolation from adult values. To determine vitamin B-6 requirements and to provide recommendations for intakes, vitamin B-6 intake, nutritional status and anthropometry were studied in 134 healthy adolescents (63 boys and 71 girls) aged 13-15 y in Tainan, Taiwan. Direct and indirect vitamin B 6 indicators were measured in plasma, erythrocytes and urine. The anthropometric data of the adolescents in this study were similar to those of the first Nutrition and Health Survey in Taiwan (NAHSIT), conducted from 1993 to 1996, showing the normal growth and development of this adolescent group. All subjects had plasma pyridoxal-5'-phosphate (PLP) concentrations > or = 20 nmol/L, indicating an adequate vitamin B-6 status. The mean dietary vitamin B-6 intakes of boys and girls were 1.04 +/- 0.24 and 0.83 +/- 0.26 mg/d, respectively. Vitamin B-6 status indicators, including plasma PLP, erythrocyte alanine activity coefficient (EALT-AC), aspartate aminotransferase activity coefficient (EAST-AC) and urinary 4-pyridoxic acid (4-PA), were correlated with vitamin B-6 intake (r = 0.84, -0.84, -0.77 and 0.86, respectively, P < 0.01). Adequate values of plasma PLP (> or = 20 nmol/L), EALT-AC (<1.25), EAST-AC (<1.8) and urinary 4-PA (>3.0 micromol/d) were used to determine the EAR according to the Dietary Reference Intake committee methodology. The present study suggests that vitamin B-6 EAR (RDA) for adolescent boys and girls aged 13-15 y are 1.07 (1.28) and 0.90 (1.08) mg/d, respectively. PMID- 14519810 TI - Dietary (+)-catechin and BHT markedly increase alpha-tocopherol concentrations in rats by a tocopherol-omega-hydroxylase-independent mechanism. AB - The effects of dietary (+)-catechin (CAT) and BHT on plasma and tissue concentrations of alpha-tocopherol (alpha-T), gamma-tocopherol (gamma-T) and cholesterol (C) were studied in male Sprague-Dawley rats. The rats were fed the compounds during a 4-wk period at concentrations of 2 g/kg in standardized diets, low but adequate in vitamin E, with 2 g/kg cholesterol. The CAT-regimen did not affect weight gain, feed intake or organ weights. BHT did not affect feed intake but lowered the body weight and the amount of liver lipids and increased the weights of livers and lungs relative to the body weight. Rats consuming CAT had 2.5-3.5-fold increased plasma, liver and lung alpha-T concentrations, but C concentrations remained unchanged. BHT-feeding resulted in 2.4- and 1.7-fold elevation in alpha-T but approximately 50% decrease in gamma-T concentrations in blood plasma and liver, respectively. BHT also lowered total C in the liver without affecting the concentration of C in the liver lipids. To investigate whether the alpha-T-sparing action of the studied compounds was due to the inhibition of tocopherol-omega-hydroxylase, HepG2 cells were incubated with CAT or BHT in the presence of delta-tocopherol (delta-T) and the 3'- and 5'-delta carboxychromanol metabolites in the media were analyzed by GC/MS. Neither CAT nor BHT inhibited tocopherol-omega-hydroxylase activity in hepatocyte cultures; CAT was also inactive in a rat microsomal assay. In conclusion, both dietary CAT and BHT markedly increased alpha-T concentrations in plasma and organs of Sprague Dawley rats by a mechanism that apparently does not involve inhibition of tocopherol-omega-hydroxylase, a key enzyme in tocopherol catabolism. PMID- 14519811 TI - Total fumonisins are reduced in tortillas using the traditional nixtamalization method of mayan communities. AB - Fumonisin B1 (FB1) is a maize mycotoxin. In tortilla preparation, maize is treated with lime (nixtamalization), producing hydrolyzed FB1 (HFB1) due to loss of the tricarballylic acid side chains. This study determined the following: 1) whether nixtamalization by Mayan communities reduces total fumonisins, and 2) the steps in the process at which reduction occurs. Tortillas prepared by the traditional process contained FB1, FB2 and FB3 and their hydrolyzed counterparts. There were equimolar amounts of FB1 and HFB1 in the tortillas, but the total fumonisins were reduced 50%. The total FB1 plus HFB1 in the residual lime water and water washes of the nixtamal accounted for 50% of the total FB1 in the uncooked maize. HFB1 and FB1 were present in a 1:1 mol/L ratio in the water washes of the nixtamal, the masa dough and the cooked tortillas, whereas the ratio of HFB1:FB1 in lime water after steeping was 21. Water washes contained 11% of the FB1 that was in the uncooked maize. The results show that the traditional method reduced the total fumonisins in tortillas and reduced the sphinganine elevation (a biomarker closely correlated with fumonisin toxicity) in cells treated with extracts of tortillas compared with cells treated with extracts of contaminated maize. PMID- 14519812 TI - Conjugated linoleic acid ameliorates viral infectivity in a pig model of virally induced immunosuppression. AB - We investigated the cellular and molecular immunoregulatory actions of conjugated linoleic acid (CLA) of relevance to viral disease pathogenesis and antiviral responses. To test the hypothesis that CLA ameliorates viral disease, we developed a viral challenge model by infecting pigs with type-2 porcine circovirus (PCV2). After 42 d of dietary supplementation with either soybean oil (n = 16) or CLA (n = 16), half of the pigs in each group were challenged with PCV2. We examined the effect of CLA on the development of lesions (i.e., lymphoid depletion and pneumonia) and observed the kinetics of the immune responses against PCV2. The viral infection depleted immature B cells (IgM+SWC3+) and favored proapoptotic mRNA expression profiles [i.e., suppressed B-cell leukemia/lymphoma-xl (Bcl-xl) and stimulated Bcl-2 homologous antagonist/killer (Bak)] in the external inguinal lymph nodes. B-cell depletion was more accentuated in pigs fed the control diet, whereas interleukin (IL)-2 mRNA expression was downregulated. Histopathological examination of the lungs revealed that the interstitial pneumonia tended to be more severe in infected pigs fed the control diet, which were also affected by growth retardation. CD8+ T cells were the primary cellular targets of CLA action in peripheral blood (CD8+CD29low and CD8+CD45RC+) and thymus (CD8+ and CD4+CD8+). CLA interacted with PCV2 to increase the proliferation of CD8+ T cells and to suppress PCV2-specific interferon (IFN) gamma production in CD4+ T cells. At the molecular level, these cellular immunoregulatory properties were associated with differential patterns of peroxisome proliferator-activated receptor (alpha and gamma) mRNA expression between diets in virally infected pigs. PMID- 14519813 TI - Perinatal iron deficiency alters the neurochemical profile of the developing rat hippocampus. AB - Cognitive deficits in human infants at risk for gestationally acquired perinatal iron deficiency suggest involvement of the developing hippocampus. To understand the plausible biological explanations for hippocampal injury in perinatal iron deficiency, a neurochemical profile of 16 metabolites in the iron-deficient rat hippocampus was evaluated longitudinally by 1H NMR spectroscopy at 9.4 T. Metabolites were quantified from an 11-24 microL volume centered in the hippocampus in 18 iron-deficient and 16 iron-sufficient rats on postnatal day (PD) 7, PD10, PD14, PD21 and PD28. Perinatal iron deficiency was induced by feeding the pregnant dam an iron-deficient diet from gestational d 3 to PD7. The brain iron concentration of the iron-deficient group was 60% lower on PD7 and 19% lower on PD28 (P < 0.001 each). The concentration of 12 of the 16 measured metabolites changed over time between PD7 and PD28 in both groups (P < 0.001 each). Compared with the iron-sufficient group, phosphocreatine, glutamate, N acetylaspartate, aspartate, gamma-aminobutyric acid, phosphorylethanolamine and taurine concentrations, and the phosphocreatine/creatine ratio were elevated in the iron-deficient group (P < 0.02 each). These neurochemical alterations suggest persistent changes in resting energy status, neurotransmission and myelination in perinatal iron deficiency. An altered neurochemical profile of the developing hippocampus may underlie some of the cognitive deficits observed in human infants with perinatal iron deficiency. PMID- 14519814 TI - Behavioral responses are altered in piglets with decreased frontal cortex docosahexaenoic acid. AB - Docosahexaenoic acid [22:6(n-3)] is required in large amounts for membrane lipid synthesis during brain growth. The functional importance of differences in dietary fatty acid intakes that alter brain 22:6(n-3), however, is not well understood. We used a dietary approach to manipulate 22:6(n-3) in piglet brain and assessed the effects on behavior and change in behavior on an elevated plus maze after administration of L-dihydroxyphenylalanine (L-Dopa) or sulperide, a dopamine D2 receptor blocker. Piglets were fed 1.2% energy 18:2(n-6) and 0.05% energy 18:3(n-3) (low PUFA), or 10.7% energy 18:2(n-6), 1.1% energy 18:3(n-3), 0.3% energy 20:4(n-6) and 0.3% energy 22:6(n-3) (high PUFA) from 1 d of age and behavior assessed at 18-22 d of age. At 30 d of age, frontal cortex dopamine, and phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidyethanolamine (PE) and phosphatidylinositol (PI) fatty acids were quantified. Piglets fed the low PUFA diet had fewer arm entries on the maze than piglets fed the high PUFA diet, P = 0.02. L-Dopa increased the open (P = 0.005) and closed (P = 0.04) arm entries by piglets fed the low PUFA diet. Behavior did not differ between piglets fed the low and high PUFA diets when given L-Dopa. Frontal cortex PC, PS and PE 22:6(n-3) was lower and 22:5(n-6) was higher in piglets fed the low compared with the high PUFA diet, P < 0.01. Our work establishes the neonatal piglet as a model with which to study the behavioral effects of diet-induced changes in brain 22:6(n-3), and provides functional evidence that brain 22:6(n-3) is important in central dopamine metabolism. PMID- 14519815 TI - The J-shaped effect of coffee consumption on the risk of developing acute coronary syndromes: the CARDIO2000 case-control study. AB - The effect of coffee consumption on cardiovascular disease has been debated for many years. In this work, we evaluated the association between coffee consumption and the risk of developing acute coronary syndromes, based on a random sample of 848 patients with their first coronary heart disease event and 1078 frequency matched controls with no cardiovascular disease in their medical history, from the entire country. The multivariate analysis raises a J-shaped association between the risk of developing acute coronary syndromes and the quantity of coffee consumed per day. In particular, the odds ratios for moderate (<300 mL/d), heavy (300-600 mL/d), and very heavy (>600 mL/d), consumption, relative to no consumption, were 0.69 (95% CI, 0.50-0.86), 1.56 (95% CI, 1.10-2.34) and 3.10 (95% CI, 1.82-5.26), respectively, after controlling for the presence of hypertension, hypercholesterolemia, diabetes mellitus, family history of premature coronary heart disease, physical activity status, smoking habits, BMI, alcohol consumption, triglycerides, consumption of several food items, depression scale score and education status. The suggested J-shaped association between coffee consumption and the risk of developing acute coronary syndromes may partially explain the conflicting results from other studies in the past. PMID- 14519817 TI - Low versus high meat diets: effects on calcium metabolism. PMID- 14519816 TI - Dietary folate and depressive symptoms are associated in middle-aged Finnish men. AB - Several cross-sectional studies have focused on the low blood folate levels of depressed patients. However, no published studies have examined the association between dietary folate and current symptoms of depression in a general population. We investigated the association between dietary folate, cobalamin, pyridoxine and riboflavin and current symptoms of depression in a cross-sectional general population study. We recruited 2682 men aged between 42 and 60 y from eastern Finland. Those who had a previous history of psychiatric disorder were excluded (n = 146, 5.6% of the cohort). Depressive symptoms were assessed with the 18-item Human Population Laboratory Depression Scale. Those who scored 5 or more at baseline were considered to have elevated depressive symptoms (n = 228, 9.3% of the cohort). The participants were grouped into thirds according to their dietary folate intake. Those in the lowest third of energy-adjusted folate intake had a higher risk of being depressed [odds ratio (OR) 1.67, 95% CI = 1.19-2.35, P = 0.003] than those in the highest folate intake third. This increased risk remained significant after adjustment for smoking habits, alcohol consumption, appetite, BMI, marital status, education, adulthood socioeconomic status and total fat consumption (OR = 1.46, 95% CI = 1.01-2.12, P = 0.044). There were no associations between the intake of cobalamin, pyridoxine or riboflavin, and depression. These results indicate that nutrition may have a role in the prevention of depression. PMID- 14519818 TI - High meat diet, acid-base status and calcium retention. PMID- 14519820 TI - Introduction to the proceedings of the Third International Scientific Symposium on Tea and Human Health: Role of Flavonoids in the Diet. PMID- 14519821 TI - Proceedings of the 3rd International Scientific Symposium on Tea and Human Health: Role of Flavonoids in the Diet. September 23, 2002. PMID- 14519822 TI - Overview of dietary flavonoids: nomenclature, occurrence and intake. AB - Flavonoids and their polymers constitute a large class of food constituents, many of which alter metabolic processes and have a positive impact on health. Flavonoids are a subclass of polyphenols. They generally consist of two aromatic rings, each containing at least one hydroxyl, which are connected through a three carbon "bridge" and become part of a six-member heterocyclic ring. The flavonoids are further divided into subclasses based on the connection of an aromatic ring to the heterocyclic ring, as well as the oxidation state and functional groups of the heterocyclic ring. Within each subclass, individual compounds are characterized by specific hydroxylation and conjugation patterns. Many flavonoids in foods also occur as large molecules (tannins). These include condensed tannins (proanthocyanidins), derived tannins and hydrolysable tannins. For proanthocyanidins, three subclasses (15 characterized) have been identified in foods. Monomers are connected through specific carbon-carbon and ether linkages to form polymers. Derived tannins are formed during food handling and processing, and found primarily in black and oolong teas. Flavonoids are widely distributed in nature, albeit not uniformly. As a result, specific groups of foods are often rich sources of one or more subclasses of these polyphenols. The polyphenolic structure of flavonoids and tannins renders them quite sensitive to oxidative enzymes and cooking conditions. Scientists in several countries have estimated intakes of a few subclasses of flavonoids from limited food composition databases. These observations suggest large differences in consumption, due in part to cultural and food preferences among populations of each country. PMID- 14519823 TI - Metabolism of tea flavonoids in the gastrointestinal tract. AB - There is considerable interest in the bioavailability of flavan-3-ols such as tea catechins and their bioactivity in vivo. Although flavanols such as catechin and epicatechin have long been characterized as powerful antioxidants in vitro, evidence suggests that these compounds undergo significant metabolism and conjugation during absorption in the small intestine and in the colon. In the small intestine these modifications lead primarily to the formation of glucuronide conjugates that are more polar than the parent flavanol and are marked for renal excretion. Other phase II processes lead to the production of O methylated forms that have reduced antioxidant potential via the methylation of the B-ring catechol. Significant modification of flavanols also occurs in the colon where the resident microflora degrade them to smaller phenolic acids, some of which may be absorbed. Cell, animal and human studies have confirmed such metabolism by the detection of flavanol metabolites in the circulation and tissues. This review will highlight the major sites of flavanol metabolism in the gastrointestinal tract and the processes that give rise to potential bioactive forms of flavan-3-ols in vivo. PMID- 14519824 TI - Mechanisms of cancer prevention by tea constituents. AB - Consumption of tea (Camellia sinensis) has been suggested to prevent cancer, heart disease and other diseases. Animal studies have shown that tea and tea constituents inhibit carcinogenesis of the skin, lung, oral cavity, esophagus, stomach, liver, prostate and other organs. In some studies, the inhibition correlated with an increase in tumor cell apoptosis and a decrease in cell proliferation. Studies with human cancer cell lines have demonstrated that epigallocatechin-3-gallate (EGCG), a major tea polyphenol, inhibits mitogen activated protein kinases, cyclin-dependent kinases, growth factor-related cell signaling, activation of activator protein 1 (AP-1) and nuclear factor kappaB (NFkappaB), topoisomerase I and matrix metalloproteinases as well as other potential targets. Although some studies report effects of EGCG at submicromolar levels, most experiments require concentrations of >10 or 20 micromol/L to demonstrate the effect. In humans, tea polyphenols undergo glucuronidation, sulfation, methylation, and ring fission. The peak plasma concentration of EGCG is approximately 1 micromol/L. The possible relevance of each of the proposed mechanisms to human cancer prevention is discussed in light of current bioavailability data for tea polyphenols and the potential limitations of animal models of carcinogenesis. Such discussion, it is hoped, will clarify some misunderstandings of cancer prevention by tea and stimulate new research efforts. PMID- 14519825 TI - Tea and cancer prevention: studies in animals and humans. AB - The role of tea in protection against cancer has been supported by ample evidence from studies in cell culture and animal models. However, epidemiological studies have generated inconsistent results, some of which associated tea with reduced risk of cancer, whereas others found that tea lacks protective activity against certain human cancers. These results raise questions about the actual role of tea in human cancer that needs to be addressed. This article is intended to provide a better perspective on this controversy by summarizing the laboratory studies in animals and humans with emphasis on animal tumor bioassays on skin, lung, mammary glands and colon, and the molecular and cellular mechanisms affected by tea. Finally, a recent small pilot intervention study with green tea in smokers is presented. PMID- 14519827 TI - Antioxidant effects of tea: evidence from human clinical trials. AB - Tea remains the most consumed drink in the world after water, well ahead of coffee, beer, wine and carbonated soft drinks. An accumulated number of population studies suggests that consumption of green and black tea beverages may bring positive health effects (1). One hypothesis explaining such effects is that the high levels of flavonoids in tea can protect cells and tissues from oxidative damage by scavenging oxygen-free radicals. Chemically, the flavonoids found in green and black tea are very effective radical scavengers. The tea flavonoids may therefore be active as antioxidants in the digestive tract or in other tissues after uptake. A substantial number of human intervention studies with green and black tea demonstrates a significant increase in plasma antioxidant capacity in humans approximately 1 h after consumption of moderate amounts of tea (1-6 cups/d). There are initial indications that the enhanced blood antioxidant potential leads to reduced oxidative damage to macromolecules such as DNA and lipids. However, the measurement of oxidative damage through biomarkers needs to be further established. In conclusion, tea flavonoids are potent antioxidants that are absorbed from the gut after consumption. Tea consumption consistently leads to a significant increase in the antioxidant capacity of the blood. Beneficial effects of increased antioxidant capacity in the body may be the reduction of oxidative damage to important biomolecules. The scientific support is strongest for the protection of DNA from oxidative damage after black or green tea consumption. However, the quality of the studies now available is insufficient to draw firm conclusions. Therefore, further evidence from human intervention studies is required. PMID- 14519826 TI - Antioxidant activity of tea polyphenols in vivo: evidence from animal studies. AB - Tea is particularly rich in polyphenols, including catechins, theaflavins and thearubigins, which are thought to contribute to the health benefits of tea. Tea polyphenols act as antioxidants in vitro by scavenging reactive oxygen and nitrogen species and chelating redox-active transition metal ions. They may also function indirectly as antioxidants through 1) inhibition of the redox-sensitive transcription factors, nuclear factor-kappaB and activator protein-1; 2) inhibition of "pro-oxidant" enzymes, such as inducible nitric oxide synthase, lipoxygenases, cyclooxygenases and xanthine oxidase; and 3) induction of phase II and antioxidant enzymes, such as glutathione S-transferases and superoxide dismutases. The fact that catechins are rapidly and extensively metabolized emphasizes the importance of demonstrating their antioxidant activity in vivo. Animal studies offer a unique opportunity to assess the contribution of the antioxidant properties of tea and tea polyphenols to the physiological effects of tea administration in different models of oxidative stress. Most promising are the consistent findings in animal models of skin, lung, colon, liver and pancreatic cancer that tea and tea polyphenol administration inhibit carcinogen induced increases in the oxidized DNA base, 8-hydroxy-2'-deoxyguanosine. In animal models of atherosclerosis, green and black tea administration has resulted in modest improvements in the resistance of lipoproteins to ex vivo oxidation, although limited data suggest that green tea or green tea catechins inhibit atherogenesis. To determine whether tea polyphenols act as effective antioxidants in vivo, future studies in animals and humans should employ sensitive and specific biomarkers of oxidative damage to lipids, proteins and DNA. PMID- 14519828 TI - Tea consumption and cardiovascular disease: effects on endothelial function. AB - Epidemiological studies suggest that tea consumption is associated with reduced cardiovascular disease risk, but the mechanisms for these observations have remained uncertain. In recent years, it has become apparent that the endothelium plays a central role in the regulation of vascular homeostasis and that endothelial dysfunction contributes to the pathogenesis and clinical expression of cardiovascular disease. This article reviews the evidence that human tea consumption has a beneficial effect on the vascular endothelium and the clinical implications of these findings. PMID- 14519830 TI - Effect of increased tea consumption on oxidative DNA damage among smokers: a randomized controlled study. AB - Tea drinking has been associated with decreased occurrence of cancer and heart disease. One potential mechanism for these findings is the strong antioxidant effect of tea polyphenols. A phase II randomized controlled tea intervention trial was designed to study the effect of high consumption (4 cups/d) of decaffeinated green or black tea on oxidative DNA damage as measured by urinary 8 hydroxydeoxyguanosine (8-OHdG) among smokers over a 4-mo period. A total of 143 heavy smokers, aged 18-79 y, were randomized to drink either green or black tea or water. Levels of plasma and urinary catechins and urinary 8-OHdG were measured monthly. A total of 133 of 143 smokers completed the 4-mo intervention. Multiple linear regression models were used to estimate the main effects and interaction effect of green and black tea consumption on creatinine-adjusted urinary 8-OHdG, with or without adjustment for potential confounders. Plasma and urinary levels of catechins rose significantly in the green tea group compared with the other two groups. Assessment of urinary 8-OHdG after adjustment for baseline measurements and other potential confounders revealed a highly significant decrease in urinary 8-OHdG (-31%) after 4 mo of drinking decaffeinated green tea (P = 0.002). No change in urinary 8-OHdG was seen among smokers assigned to the black tea group. These data suggest that regular green tea drinking might protect smokers from oxidative damages and could reduce cancer risk or other diseases caused by free radicals associated with smoking. PMID- 14519829 TI - Black tea consumption reduces total and LDL cholesterol in mildly hypercholesterolemic adults. AB - Despite epidemiological evidence that tea consumption is associated with the reduced risk of coronary heart disease, experimental studies designed to show that tea affects oxidative stress or blood cholesterol concentration have been unsuccessful. We assessed the effects of black tea consumption on lipid and lipoprotein concentrations in mildly hypercholesterolemic adults. Tea and other beverages were included in a carefully controlled weight-maintaining diet. Five servings/d of tea were compared with a placebo beverage in a blinded randomized crossover study (7 men and 8 women, consuming a controlled diet for 3 wk/treatment). The caffeine-free placebo was prepared to match the tea in color and taste. In a third period, caffeine was added to the placebo in an amount equal to that in the tea. Five servings/d of tea reduced total cholesterol 6.5%, LDL cholesterol 11.1%, apolipoprotein B 5% and lipoprotein(a) 16.4% compared with the placebo with added caffeine. Compared with the placebo without added caffeine, total cholesterol was reduced 3.8% and LDL cholesterol was reduced 7.5% whereas apolipoprotein B, Lp(a), HDL cholesterol, apolipoprotein A-I and triglycerides were unchanged. Plasma oxidized LDL, F2-isoprostanes, urinary 8 hydroxy-2'-deoxyguanosine, ex vivo ferric ion reducing capacity and thiobarbituric acid reactive substances in LDL were not affected by tea consumption compared with either placebo. Thus, inclusion of tea in a diet moderately low in fat reduces total and LDL cholesterol by significant amounts and may, therefore, reduce the risk of coronary heart disease. Tea consumption did not affect antioxidant status in this study. PMID- 14519831 TI - The epidemiology of tea consumption and colorectal cancer incidence. AB - This manuscript provides a brief synopsis of 30 studies aimed at examining tea consumption as a factor in the incidence of colon and rectal cancers. The 30 papers examine populations in 12 countries and provide data on consumption of both black and green tea. These studies do not provide consistent evidence to support the theory from animal studies and basic research that tea is a potent chemopreventive agent. Details of the studies are presented, and the potential impact of measurement error, publication bias, the form of tea consumed, the appropriateness of the outcomes studied and the adjustment of confounders related to both tea consumption and risk of colorectal cancer or polyps in various countries are explored. In general, the data are not more consistent for green than for black tea. Particularly with green tea, the doses consumed do get into a perceived protective range in a significant subset of the population. A negative association is stronger in observational epidemiologic studies of rectal cancer than in colon cancer. There is no consistent adjustment for important potential confounders of any tea relationship, such as coffee and alcohol consumption and physical activity levels. Finally, the assessment of tea in most of these studies was based on a single question and therefore may have significant measurement error compared with more recent studies specifically aimed at assessing tea consumption. PMID- 14519833 TI - Festschrift edition of the Journal of Medical Ethics in honour of Raanan Gillon. PMID- 14519834 TI - Four scenarios. PMID- 14519835 TI - Methods and principles in biomedical ethics. AB - The four principles approach to medical ethics plus specification is used in this paper. Specification is defined as a process of reducing the indeterminateness of general norms to give them increased action guiding capacity, while retaining the moral commitments in the original norm. Since questions of method are central to the symposium, the paper begins with four observations about method in moral reasoning and case analysis. Three of the four scenarios are dealt with. It is concluded in the "standard" Jehovah's Witness case that having autonomously chosen the authority of his religious institution, a Jehovah's Witness has a reasonable basis on which to refuse a recommended blood transfusion. The author's view of the child of a Jehovah's Witness scenario is that it is morally required not merely permitted-to overrule this parental refusal of treatment. It is argued in the selling kidneys for transplantation scenario that a fair system of regulating and monitoring would be better than the present system which the author believes to be a shameful failure. PMID- 14519836 TI - Applying the four principles. AB - Gillon is correct that the four principles provide a sound and useful way of analysing moral dilemmas. As he observes, the approach using these principles does not provide a unique solution to dilemmas. This can be illustrated by alternatives to Gillon's own analysis of the four case scenarios. In the first scenario, a different set of factual assumptions could yield a different conclusion about what is required by the principle of beneficence. In the second scenario, although Gillon's conclusion is correct, what is open to question is his claim that what society regards as the child's best interest determines what really is in the child's best interest. The third scenario shows how it may be reasonable for the principle of beneficence to take precedence over autonomy in certain circumstances, yet like the first scenario, the ethical conclusion relies on a set of empirical assumptions and predictions of what is likely to occur. The fourth scenario illustrates how one can draw different conclusions based on the importance given to the precautionary principle. PMID- 14519837 TI - Juggling law, ethics, and intuition: practical answers to awkward questions. AB - The eclectic problem solving methodology used by the British Medical Association (BMA) is described in this paper. It has grown from the daily need to respond to doctors' practical queries and incorporates reference to law, traditional professional codes, and established BMA policies-all of which must be regularly assessed against the benchmark of contemporary societal expectations. The two Jehovah's Witness scenarios are analysed, using this methodology and in both cases the four principles solution is found to concur with that of the BMA's approach. The author's overall conclusion is that although the BMA resorts to a lengthier list of things to consider, the solutions that emerge are often likely to coincide with the four principles approach. PMID- 14519838 TI - Principlism and communitarianism. AB - The decline in the interest in ethical theory is first outlined, as a background to the author's discussion of principlism. The author's own stance, that of a communitarian philosopher, is then described, before the subject of principlism itself is addressed. Two problems stand in the way of the author's embracing principlism: its individualistic bias and its capacity to block substantive ethical inquiry. The more serious problem the author finds to be its blocking function. Discussing the four scenarios the author finds that the utility of principlism is shown in the two scenarios about Jehovah's Witnesses but that when it comes to selling kidneys for transplantation and germline enhancement, principlism is of little help. PMID- 14519839 TI - The virtues (and vices) of the four principles. AB - Despite tendencies to compete for a prime place in moral theory, neither virtue ethics nor the four principles approach should claim to be superior to, or logically prior to, the other. Together they provide a more adequate account of the moral life than either can offer on its own. The virtues of principlism are clarity, simplicity and (to some extent) universality. These are well illustrated by Ranaan Gillon's masterly analysis of the cases he has provided. But the vices of this approach are the converse of its virtues: neglect of emotional and personal factors, oversimplification of the issues, and excessive claims to universality. Virtue ethics offers a complementary approach, providing insights into moral character, offering a blend of reason and emotion, and paying attention to the context of decisions. The cases provided can be more adequately understood if we combine the approaches. Both should foster the virtues of humility and magnanimity. PMID- 14519840 TI - A virtue ethics approach to moral dilemmas in medicine. AB - Most moral dilemmas in medicine are analysed using the four principles with some consideration of consequentialism but these frameworks have limitations. It is not always clear how to judge which consequences are best. When principles conflict it is not always easy to decide which should dominate. They also do not take account of the importance of the emotional element of human experience. Virtue ethics is a framework that focuses on the character of the moral agent rather than the rightness of an action. In considering the relationships, emotional sensitivities, and motivations that are unique to human society it provides a fuller ethical analysis and encourages more flexible and creative solutions than principlism or consequentialism alone. Two different moral dilemmas are analysed using virtue ethics in order to illustrate how it can enhance our approach to ethics in medicine. PMID- 14519841 TI - In praise of unprincipled ethics. AB - In this paper a plea is made for an unprincipled approach to biomedical ethics, unprincipled of course just in the sense that the four principles are neither the start nor the end of the process of ethical reflection. While the four principles constitute a useful "checklist" approach to bioethics for those new to the field, and possibly for ethics committees without substantial ethical expertise approaching new problems, it is an approach which if followed by the bioethics community as a whole would, the author believes, lead to sterility and uniformity of approach of a quite mindbogglingly boring kind. Moreover, much of bioethics is not concerned with identifying the principles or values appropriate to a particular issue, but rather involves analysing the arguments that are so often already in play and which present themselves as offering solutions in one direction or another. Here, as I try to show in discussion of these four scenarios, the principles allow massive scope in interpretation and are, frankly, not wonderful as a means of detecting errors and inconsistencies in argument. PMID- 14519842 TI - Ethics needs principles--four can encompass the rest--and respect for autonomy should be "first among equals". AB - It is hypothesised and argued that "the four principles of medical ethics" can explain and justify, alone or in combination, all the substantive and universalisable claims of medical ethics and probably of ethics more generally. A request is renewed for falsification of this hypothesis showing reason to reject any one of the principles or to require any additional principle(s) that can't be explained by one or some combination of the four principles. This approach is argued to be compatible with a wide variety of moral theories that are often themselves mutually incompatible. It affords a way forward in the context of intercultural ethics, that treads the delicate path between moral relativism and moral imperialism. Reasons are given for regarding the principle of respect for autonomy as "first among equals", not least because it is a necessary component of aspects of the other three. A plea is made for bioethicists to celebrate the approach as a basis for global moral ecumenism rather than mistakenly perceiving and denigrating it as an attempt at global moral imperialism. PMID- 14519843 TI - Toxin-induced resistance in Bacillus anthracis lethal toxin-treated macrophages. AB - In the current study, we show that macrophages adaptively resist anthrax lethal toxin (LT) through a toxin-activated process termed toxin-induced resistance (TIR). TIR was triggered by pretreatment of RAW 264.7 or J774A.1 macrophages with a low dose of LT for at least 6 h, which resulted in resistance to high doses of LT for 96 h. Activation of TIR required functional toxin, because LT subunits, mutants, and heat-inactivated toxin were unable to trigger resistance. TIR macrophages were not altered in toxin receptor levels or cell cycle profiles. Treatment of TIR macrophages with high doses of LT resulted in a sustained decline in full-length mitogen-activated protein kinase kinase 2, a known target of lethal factor, and a marked reduction in diphosphorylated extracellular response kinases 1,2 for 24 h. However, despite the sustained loss of full-length mitogen-activated protein kinase kinase 2, by 48 h, TIR macrophages regained diphosphorylated extracellular response kinases 1,2, suggesting an adaptation led to recovery of this signaling pathway. TIR macrophages were also able to maintain normal levels of ubiquitinylated proteins, whereas sensitive cells show a rapid reduction in ubiquitin-modified proteins before cell death, indicating a possible alteration in proteasome activity contributed to resistance. These results provide a paradigm for toxin-cell interactions and suggest macrophages are capable of adapting to and tolerating toxic doses of LT. PMID- 14519844 TI - Divergent retroviral late-budding domains recruit vacuolar protein sorting factors by using alternative adaptor proteins. AB - The release of enveloped viruses from infected cells often requires a virally encoded activity, termed a late-budding domain (L domain), encoded by essential PTAP, PPXY, or YPDL sequence motifs. PTAP-type L domains recruit one of three endosomal sorting complexes required for transport (ESCRT-I). However, subsequent events in viral budding are poorly defined, and neither YPDL nor PPXY-type L domains require ESCRT-I. Here, we show that ESCRT-I and other class E vacuolar protein sorting (VPS) factors are linked by a complex series of protein-protein interactions. In particular, interactions between ESCRT-I and ESCRT-III are bridged by AIP-1/ALIX, a mammalian orthologue of the yeast class E VPS factor, Bro1. Expression of certain ESCRT-III components as fusion proteins induces a late budding defect that afflicts all three L-domain types, suggesting that ESCRT III integrity is required in a general manner. Notably, the prototype YPDL-type L domain encoded by equine infectious anemia virus (EIAV) acts by recruiting AIP 1/ALIX and expression of a truncated form of AIP-1/ALIX or small interfering RNA induced AIP-1/ALIX depletion specifically inhibits EIAV YPDL-type L-domain function. Overall, these findings indicate that L domains subvert a subset of class E VPS factors to mediate viral budding, some of which are required for each of the L-domain types, whereas others apparently act as adaptors to physically link specific L-domain types to the class E VPS machinery. PMID- 14519845 TI - Patterning of the cardiac outflow region in Drosophila. AB - Specification of bilateral cardiac primordia and formation of the linear heart tube are highly conserved from Drosophila to humans. However, subsequent heart morphogenesis involving nonmesodermal neural crest cells was thought to be specific for vertebrates. Here, we provide evidence that a group of nonmesodermal cells that we have named heart-anchoring cells (HANCs) contribute to heart morphogenesis in Drosophila. We show that the homeobox genes ladybird (lb) known to be involved in diversification of cardiac precursors are expressed in HANCs and required for their specification. Interestingly, the HANCs selectively contact the anterior cardiac cells, which express lb as well. Direct interaction between HANCs and cardiac cells is assisted by a pair of cardiac outflow muscles (COMs), each of which selectively attaches to both the lb-expressing cardiac cells and HANCs. COM muscles seem to ensure ventral bending of the heart tip and together with HANCs determine the spatial positioning of the cardiac outflow region. Experimentally depleted cardiac lb expression leads to the disruption of the contact between the tip of the heart and either the COM muscles or the HANC cells, indicating a pivotal morphogenetic role for the lb expression within the heart. PMID- 14519846 TI - Predicting aberrant CpG island methylation. AB - Epigenetic silencing associated with aberrant methylation of promoter region CpG islands is one mechanism leading to loss of tumor suppressor function in human cancer. Profiling of CpG island methylation indicates that some genes are more frequently methylated than others, and that each tumor type is associated with a unique set of methylated genes. However, little is known about why certain genes succumb to this aberrant event. To address this question, we used Restriction Landmark Genome Scanning to analyze the susceptibility of 1,749 unselected CpG islands to de novo methylation driven by overexpression of DNA cytosine-5 methyltransferase 1 (DNMT1). We found that although the overall incidence of CpG island methylation was increased in cells overexpressing DNMT1, not all loci were equally affected. The majority of CpG islands (69.9%) were resistant to de novo methylation, regardless of DNMT1 overexpression. In contrast, we identified a subset of methylation-prone CpG islands (3.8%) that were consistently hypermethylated in multiple DNMT1 overexpressing clones. Methylation-prone and methylation-resistant CpG islands were not significantly different with respect to size, C+G content, CpG frequency, chromosomal location, or promoter association. We used DNA pattern recognition and supervised learning techniques to derive a classification function based on the frequency of seven novel sequence patterns that was capable of discriminating methylation-prone from methylation-resistant CpG islands with 82% accuracy. The data indicate that CpG islands differ in their intrinsic susceptibility to de novo methylation, and suggest that the propensity for a CpG island to become aberrantly methylated can be predicted based on its sequence context. PMID- 14519847 TI - Unique conformation of cancer autoantigen B23 in hepatoma: a mechanism for specificity in the autoimmune response. AB - The association of a specific autoantibody response with distinct disease phenotypes is observed in both autoimmune diseases and cancer. Although the underlying mechanisms remain unclear, it is likely that unique properties of disease-specific autoantigens expressed in the relevant target cells play a role. It has recently been observed that the majority of autoantigens targeted across the spectrum of systemic autoimmune diseases (but not nonautoantigens) are selectively cleaved by the cytotoxic lymphocyte granule protease granzyme B (GB), generating unique fragments not observed during other forms of cell death. Although susceptibility of a molecule to cleavage by GB strongly predicts autoantigen status, the significance of this association is unclear. We used hepatocellular carcinoma and the hepatocellular carcinoma autoantigen, nucleophosmin/B23, as a model system to define the unique features of disease specific autoantigens in the relevant disease microenvironment. These studies revealed a striking, selective susceptibility of B23 to cleavage by GB in extracts of neoplastic liver. The increased sensitivity of tumor B23 to proteolysis by GB was accompanied by slightly increased mobility on SDS/PAGE, altered subcellular localization, enrichment of an SDS-stable oligomeric form of B23, and recognition by a conformation-specific antibody detecting a B23 epitope ending at the GB cleavage site. In vitro studies demonstrated that this unique B23 conformation and resultant increased susceptibility to cleavage by GB arise when B23 translation is initiated at methionine-7. We propose that unique features of autoantigens in the disease-relevant microenvironment may regulate susceptibility to cleavage by GB and their selection by the specific autoimmune response. PMID- 14519848 TI - A strabismus susceptibility locus on chromosome 7p. AB - Strabismus has been known to have a significant genetic component, but the mode of inheritance and the identity of the relevant genes have been enigmatic. This paper reports linkage analysis of nonsyndromic strabismus. The principal results of this study are: (i) the demonstrated feasibility of identifying and recruiting large families in which multiple members have (or had) strabismus; (ii) the linkage in one large family of a presumptive strabismus susceptibility locus to 7p22.1 with a multipoint logarithm of odds score of 4.51 under a model of recessive inheritance; and (iii) the failure to observe significant linkage to 7p in six other multiplex families, consistent with genetic heterogeneity among families. These findings suggest that it will be possible to localize and ultimately identify strabismus susceptibility genes by linkage analysis and mutation screening of candidate genes. PMID- 14519849 TI - Alpha1E-containing Ca2+ channels are involved in synaptic plasticity. AB - Long-term potentiation (LTP) is the most prominent model for the molecular and cellular mechanisms of learning and memory. Two main forms of LTP have been distinguished. The N-methyl-D-aspartate-receptor-dependent forms of LTP have been studied most extensively, whereas much less is known about N-methyl-D-aspartate receptor-independent forms of LTP. This latter type of LTP was first described at the mossy fiber synapses in the hippocampus and subsequently at parallel fiber synapses in the cerebellum as well as at corticothalamic synapses. These presynaptic forms of LTP require a rise in the intraterminal calcium concentration, but the channel through which calcium passes has not been identified. By using pharmacological tools as well as genetic deletion, we demonstrate here that alpha1E-containing voltage-dependent calcium channels (VDCCs) shift the threshold for mossy fiber LTP. The channel is not involved in the expression mechanism, but it contributes to the calcium influx during the induction phase. Indeed, optical recordings directly show the presence and the function of alpha1E-containing VDCCs at mossy fiber terminals. Hence, a previously undescribed role for alpha1E-containing VDCCs is suggested by these results. PMID- 14519850 TI - Nuclear localization of pyrrole-imidazole polyamide-fluorescein conjugates in cell culture. AB - A series of hairpin pyrrole-imidazole polyamide-fluorescein conjugates were synthesized and assayed for cellular localization. Thirteen cell lines, representing 11 human cancers, one human transformed kidney cell line, and one murine leukemia cell line, were treated with 5 microM polyamide-fluorescein conjugates for 10-14 h, then imaged by confocal laser scanning microscopy. A conjugate containing a beta-alanine residue at the C terminus of the polyamide moiety showed no nuclear localization, whereas an analogous compound lacking the beta-alanine residue was strongly localized in the nuclei of all cell lines tested. The localization profiles of several other conjugates suggest that pyrrole-imidazole sequence and content, dye choice and position, linker composition, and molecular weight are determinants of nuclear localization. The attachment of fluorescein to the C terminus of a hairpin polyamide results in an approximately 10-fold reduction in DNA-binding affinity, with no loss of binding specificity with reference to mismatch binding sites. PMID- 14519852 TI - Soluble CD40 ligand induces beta3 integrin tyrosine phosphorylation and triggers platelet activation by outside-in signaling. AB - We earlier reported that the soluble form of the CD40 ligand (sCD40L), is involved in thrombosis by stabilizing platelet thrombi. In this article, we have determined the mechanism by which this protein affects platelet biology. Addition of sCD40L to washed platelets was found to activate the receptor function of alphaIIbbeta3 as measured by the induction of fibrinogen binding and the formation of platelet microparticles. Mutation in the KGD sequence (D117E) of sCD40L, the alphaIIbbeta3-binding domain in the N terminus of the protein resulted in a loss of the platelet-stimulatory activity of this protein. Integrilin, a alphaIIbbeta3 antagonist, but not an antibody to CD40 that blocked the ligand-binding activity, inhibited these platelet-stimulatory events. CD40-/- platelets bound fibrinogen and formed microparticles similar to WT platelets, again indicating that CD40 is not involved in sCD40L-induced platelet activation. Exposure of platelets to sCD40L, but not D117E-sCD40L-coated surfaces, induced platelet thrombi formation under arterial shear rate. sCD40L-induced platelet stimulation resulted in the phosphorylation of tyrosine-759 in the cytoplasmic domain of beta3. Platelets from the diYF mouse strain, expressing beta3 in which both cytoplasmic tyrosines are mutated to phenylalanine, were defective in sCD40L induced platelet stimulation. These data indicate that sCD40L is a primary platelet agonist and that platelet stimulation is induced by the binding of the KGD domain of sCD40L to alphaIIbbeta3, triggering outside-in signaling by tyrosine phosphorylation of beta3. PMID- 14519851 TI - Nucleation-dependent conformational conversion of the Y145Stop variant of human prion protein: structural clues for prion propagation. AB - One of the most intriguing disease-related mutations in human prion protein (PrP) is the Tyr to Stop codon substitution at position 145. This mutation results in a Gerstmann-Straussler-Scheinker-like disease with extensive PrP amyloid deposits in the brain. Here, we provide evidence for a spontaneous conversion of the recombinant polypeptide corresponding to the Y145Stop variant (huPrP23-144) from a monomeric unordered state to a fibrillar form. This conversion is characterized by a protein concentration-dependent lag phase and has characteristics of a nucleation-dependent polymerization. Atomic force microscopy shows that huPrP23 144 fibrils are characterized by an apparent periodicity along the long axis, with an average period of 20 nm. Fourier-transform infrared spectra indicate that the conversion is associated with formation of beta-sheet structure. However, the infrared bands for huPrP23-144 are quite different from those for a synthetic peptide PrP106-126, suggesting conformational non-equivalence of beta-structures in the disease-associated Y145Stop variant and a frequently used short model peptide. To identify the region that is critical for the self-seeded assembly of huPrP23-144 amyloid, experiments were performed by using the recombinant polypeptides corresponding to prion protein fragments 23-114, 23-124, 23-134, 23 137, 23-139, and 23-141. Importantly, none of the fragments ending before residue 139 showed a propensity for conformational conversion to amyloid fibrils, indicating that residues within the 138-141 region are essential for this conversion. PMID- 14519853 TI - Self-organized queuing and scale-free behavior in real escape panic. AB - Numerical investigations of escape panic of confined pedestrians have revealed interesting dynamical features such as pedestrian arch formation around an exit, disruptive interference, self-organized queuing, and scale-free behavior. However, these predictions have remained unverified because escape panic experiments with real systems are difficult to perform. For mice escaping out of a water pool, we found that for a critical sampling rate the escape behavior exhibits the predicted features even at short observation times. The mice escaped via an exit in bursts of different sizes that obey exponential and (truncated) power-law distributions depending on exit width. Oversampling or undersampling the mouse escape rate prevents the observation of the predicted features. Real systems are normally subject to unavoidable constraints arising from occupancy rate, pedestrian exhaustion, and nonrigidity of pedestrian bodies. The effect of these constraints on the dynamics of real escape panic is also studied. PMID- 14519854 TI - Avian migration phenology and global climate change. AB - There is mounting evidence that global climate change has extended growing seasons, changed distribution patterns, and altered the phenology of flowering, breeding, and migration. For migratory birds, the timing of arrival on breeding territories and over-wintering grounds is a key determinant of reproductive success, survivorship, and fitness. But we know little of the factors controlling earlier passage in long-distance migrants. Over the past 30 years in Oxfordshire, U.K., the average arrival and departure dates of 20 migrant bird species have both advanced by 8 days; consequently, the overall residence time in Oxfordshire has remained unchanged. The timing of arrival has advanced in relation to increasing winter temperatures in sub-Saharan Africa, whereas the timing of departure has advanced after elevated summer temperatures in Oxfordshire. This finding demonstrates that migratory phenology is quite likely to be affected by global climate change and links events in tropical winter quarters with those in temperate breeding areas. PMID- 14519855 TI - Origins of mitochondrial thymidine triphosphate: dynamic relations to cytosolic pools. AB - Nuclear and mitochondrial (mt) DNA replication occur within two physically separated compartments and on different time scales. Both require a balanced supply of dNTPs. During S phase, dNTPs for nuclear DNA are synthesized de novo from ribonucleotides and by salvage of thymidine in the cytosol. Mitochondria contain specific kinases for salvage of deoxyribonucleosides that may provide a compartmentalized synthesis of dNTPs. Here we investigate the source of intra-mt thymidine phosphates and their relationship to cytosolic pools by isotope-flow experiments with [3H]thymidine in cultured human and mouse cells by using a rapid method for the clean separation of mt and cytosolic dNTPs. In the absence of the cytosolic thymidine kinase, the cells (i) phosphorylate labeled thymidine exclusively by the intra-mt kinase, (ii) export thymidine phosphates rapidly to the cytosol, and (iii) use the labeled dTTP for nuclear DNA synthesis. The specific radioactivity of dTTP is highly diluted, suggesting that cytosolic de novo synthesis is the major source of mt dTTP. In the presence of cytosolic thymidine kinase dilution is 100-fold less, and mitochondria contain dTTP with high specific radioactivity. The rapid mixing of the cytosolic and mt pools was not expected from earlier data. We propose that in proliferating cells dNTPs for mtDNA come largely from import of cytosolic nucleotides, whereas intra-mt salvage of deoxyribonucleosides provides dNTPs in resting cells. Our results are relevant for an understanding of certain genetic mitochondrial diseases. PMID- 14519858 TI - Coarse-grained stochastic models for tropical convection and climate. AB - Prototype coarse-grained stochastic parametrizations for the interaction with unresolved features of tropical convection are developed here. These coarse grained stochastic parametrizations involve systematically derived birth/death processes with low computational overhead that allow for direct interaction of the coarse-grained dynamical variables with the smaller-scale unresolved fluctuations. It is established here for an idealized prototype climate scenario that, in suitable regimes, these coarse-grained stochastic parametrizations can significantly impact the climatology as well as strongly increase the wave fluctuations about an idealized climatology. PMID- 14519856 TI - Functional map of human cytomegalovirus AD169 defined by global mutational analysis. AB - Human cytomegalovirus has a complex double-stranded DNA genome of approximately 240,000 bp that contains approximately 150 ORFs likely to encode proteins, most of whose functions are not well understood. We have used an infectious bacterial artificial chromosome to introduce 413 defined insertion and substitution mutations into the human cytomegalovirus AD169 genome by random and site-directed transposon mutagenesis. Mutations were produced in all unique ORFs with a high probability of encoding proteins for which mutants have not been previously documented and in many previously characterized ORFs. The growth of selected mutants was assayed in cultured human fibroblasts, and we now recognize 41 essential, 88 nonessential, and 27 augmenting ORFs. Most essential and augmenting genes are located in the central region, and nonessential genes generally cluster near the ends of the viral genome. PMID- 14519859 TI - Does selection mold molecular networks? AB - The dissection of molecular networks vital to cellular life can provide important hints about optimal network design principles. However, these hints can become conclusive only if one can determine that natural selection has molded a network's structure. I illustrate the importance of thorough studies of network evolution with two recent examples, one from genome-scale networks, the other from small transcriptional regulation circuits. PMID- 14519857 TI - Chromatin assembly factor 1 is essential and couples chromatin assembly to DNA replication in vivo. AB - De novo chromatin assembly maintains histone density on the daughter strands in the wake of the replication fork. The heterotrimer chromatin assembly factor 1 (CAF-1) couples DNA replication to histone deposition in vitro, but is not essential for yeast cell proliferation. Depletion of CAF-1 in human cell lines demonstrated that CAF-1 was required for efficient progression through S-phase. Cells lacking CAF-1 accumulated in early and mid S-phase and replicated DNA slowly. The checkpoint kinase Chk1, but not Chk2, was phosphorylated in response to CAF-1 depletion, consistent with a DNA replication defect. CAF-1-depleted cell extracts completely lacked DNA replication-coupled chromatin assembly activity, suggesting that CAF-1 is required for efficient S-phase progression in human cells. These results indicate that, in contrast to yeast, human CAF-1 is necessary for coupling chromatin assembly with DNA replication. PMID- 14519860 TI - What multi-detector row CT parameters are best for detection of colon polyps? PMID- 14519861 TI - Receiver operating characteristic curves and their use in radiology. AB - Sensitivity and specificity are the basic measures of accuracy of a diagnostic test; however, they depend on the cut point used to define "positive" and "negative" test results. As the cut point shifts, sensitivity and specificity shift. The receiver operating characteristic (ROC) curve is a plot of the sensitivity of a test versus its false-positive rate for all possible cut points. The advantages of the ROC curve as a means of defining the accuracy of a test, construction of the ROC, and identification of the optimal cut point on the ROC curve are discussed. Several summary measures of the accuracy of a test, including the commonly used percentage of correct diagnoses and area under the ROC curve, are described and compared. Two examples of ROC curve application in radiologic research are presented. PMID- 14519862 TI - The doctor is not in: it's time for a change. PMID- 14519863 TI - Road maps for advancement of radiologic computer-aided detection in the 21st century. PMID- 14519864 TI - Longevity is the most appropriate measure of health effects of radiation. PMID- 14519865 TI - The "Healthy Worker Effect": science or prejudice? PMID- 14519866 TI - The weight of evidence does not support the suggestion that exposure to low doses of X rays increases longevity. PMID- 14519867 TI - Percutaneous vertebroplasty. AB - This review, aimed at current practitioners of vertebroplasty, highlights recent changes in patient work-up and procedural techniques that have streamlined the authors' clinical practice. Preprocedural work-up, including history, physical examination, and adjunctive imaging techniques, are discussed. Technical details are reviewed, including types of equipment, techniques of needle placement, and utility of venography. Postprocedural issues are noted, including risk of subsequent fracture after vertebroplasty, long-term outcome of cement in the vertebral body, and utility of prophylactic vertebroplasty. Finally, the current state of evidence in support of the efficacy of vertebroplasty are discussed, with particular attention to the need for ongoing clinical trials. PMID- 14519868 TI - Diffusion-weighted MR imaging with apparent diffusion coefficient and apparent diffusion tensor maps in cervical spondylotic myelopathy. AB - PURPOSE: To evaluate diffusion-weighted magnetic resonance (MR) imaging in patients with cervical spondylosis and/or myelopathy. MATERIALS AND METHODS: A multishot echo-planar imaging sequence with calculation of apparent diffusion coefficient (ADC) and apparent diffusion tensor (ADT) was applied in 36 patients with symptomatic cervical spondylosis. Diffusion-weighted images read by two neuroradiologists were compared with T2-weighted fast spin-echo images read independently by three neuroradiologists with regard to clinical status (n = 36). MR findings in a selected subgroup of 20 patients whose clinical status was confirmed by electrophysiologic examination also were compared. Sensitivity, specificity, positive predictive value, and negative predictive value of both T2 weighted imaging and diffusion-weighted imaging (ADC and ADT) were calculated and compared. RESULTS: Patients with myelopathy had abnormal ADC (17 of 21) and ADT (15 of 19) maps with increased ADC and ADT values and decreased anisotropy. For the detection of myelopathy, diffusion-weighted ADC maps had a sensitivity of 80% (17 of 21), while T2-weighted images had a sensitivity of 61% (13 of 21). The negative predictive value was 63% (seven of 11) and 60% (12 of 20) for ADC maps and T2-weighted images, respectively. Conversely, the specificity of diffusion weighted images (53%; seven of 13) was lower than that of T2-weighted images (92%; 12 of 13). In patients with myelopathy confirmed at electrophysiologic examination, the sensitivity of diffusion-weighted images increased to 92% (12 of 13) and the negative predictive value increased to 75% (three of four), while T2 weighted images had a 53% (seven of 13) sensitivity and a 50% (six of 12) negative predictive value. CONCLUSION: Diffusion weighting improved the sensitivity of imaging in cervical spondylotic myelopathy. PMID- 14519870 TI - Case 63: hepatopulmonary syndrome. PMID- 14519871 TI - Pancreatic malignancy: value of arterial, pancreatic, and hepatic phase imaging with multi-detector row CT. AB - PURPOSE: To assess the value of arterial, pancreatic, and hepatic phase imaging at multi-detector row computed tomography (CT) of the pancreas for pancreatic malignancy. MATERIALS AND METHODS: Thirty-nine patients suspected of having resectable pancreatic adenocarcinoma underwent triple-phase multi-detector row CT. Images obtained during each phase were interpreted by one radiologist who evaluated presence of tumor, vascular invasion, and flow artifacts in the superior mesenteric vein and measured attenuation of tumor, normal pancreas, aorta, and superior mesenteric vein. Results were compared with histologic, follow-up, and correlative imaging findings. RESULTS: Mean tumor-to-gland attenuation difference was greatest on images obtained in the pancreatic phase (42 HU) versus that on those obtained in the hepatic phase (35 HU) and in the arterial phase (25 HU). For tumor detection, sensitivity of the images obtained in pancreatic (0.97 [29 of 30]) and hepatic (0.93 [28 of 30]) phases was superior to that of those obtained in arterial phase (0.63 [19 of 30]) (P < or =.008). For vascular invasion detection, sensitivity of images obtained in the hepatic phase (0.83) was better than that of those obtained in the pancreatic (0.58) and arterial (0.25) phases. Images obtained in the pancreatic phase demonstrated more flow artifacts and decreased attenuation in the superior mesenteric vein, compared with the artifacts revealed on images obtained in the hepatic phase. CONCLUSION: Routine acquisition of images in the arterial phase is unnecessary for detection of pancreatic adenocarcinoma. Images of the pancreas obtained in the hepatic phase with multi-detector row CT most accurately display vascular invasion. PMID- 14519872 TI - Multi-detector row CT colonography: effect of collimation, pitch, and orientation on polyp detection in a human colectomy specimen. AB - PURPOSE: To investigate the effects of orientation, collimation, pitch, and tube current setting on polyp detection at multi-detector row computed tomographic (CT) colonography and to determine the optimal combination of scanning parameters for screening. MATERIALS AND METHODS: A colectomy specimen containing 117 polyps of different sizes was insufflated and imaged with a multi-detector row CT scanner at various collimation (1.25 and 2.5 mm), pitch (3 and 6), and tube current (50, 100, and 150 mA) settings. Two-dimensional multiplanar reformatted images and three-dimensional endoluminal surface renderings from the 12 resultant data sets were examined by one observer for the presence and conspicuity of polyps. The results were analyzed with Poisson regression and logistic regression to determine the effects of scanning parameters and of specimen orientation on polyp detection. RESULTS: The percentage of polyps that were detected significantly increased when collimation (P =.008) and table feed (P =.03) were decreased. Increased tube current resulted in improved detection only of polyps with a diameter of less than 5 mm. Polyps of less than 5 mm were optimally depicted with a collimation of 1.25 mm, a pitch of 3, and a tube current setting of 150 mA; polyps with a diameter greater than 5 mm were adequately depicted with 1.25-mm collimation and with either pitch setting and any of the three tube current settings. Small polyps in the transverse segment (positioned at a 90 degrees angle to the z axis of scanning) were significantly less visible than those in parallel or oblique orientations (P <.001). The effective radiation dose, calculated with a Monte Carlo simulation, was 1.4-10.0 mSv. CONCLUSION: Detection of small polyps (<5 mm) with multi-detector row CT is highly dependent on collimation, pitch, and, to a lesser extent, tube current. Collimation of 1.25 mm, combined with pitch of 6 and tube current of 50 mA, provides for reliable detection of polyps 5 mm or larger while limiting the effective radiation dose. Polyps smaller than 5 mm, however, may be poorly depicted with use of these settings in the transverse colon. PMID- 14519873 TI - Contrast-enhanced MR imaging of postoperative scars and VX2 carcinoma in rabbits: comparison of macromolecular contrast agent and gadopentetate dimeglumine. AB - PURPOSE: To compare the magnetic resonance (MR) imaging enhancement patterns of a blood pool contrast agent, SH L 643A, with those of gadopentetate dimeglumine in postoperative scars and VX2 carcinomas in rabbits and to compare these enhancement patterns with microvessel density in pathologic specimens. MATERIALS AND METHODS: Eighteen rabbits with experimentally induced postoperative scars (n = 12) or VX2 carcinoma (n = 6) in the thighs underwent sequential MR imaging first with gadopentetate dimeglumine and then, 24 hours later, with SH L 643A. The enhancement ratios (ie, the ratios of postcontrast to precontrast signal intensity) and the microvessel densities of postoperative scars and VX2 carcinomas were assessed. Differences were tested for by using the Mann-Whitney U and Wilcoxon signed rank tests. RESULTS: In postoperative scars, enhancement ratios were consistently lower with injection of SH L 643A than with injection of gadopentetate dimeglumine for up to 30 minutes (P <.05). In postoperative scars, mean peak enhancement ratios were 1.29 +/- 0.15 (SD) with injection of SH L 643A and 1.61 +/- 0.31 with injection of gadopentetate dimeglumine (P <.01). In VX2 carcinomas, the enhancement ratios were not significantly different with injection of SH L 643A than with injection of gadopentetate dimeglumine at all time points. The mean difference between the enhancement ratios of the VX2 carcinomas and postoperative scars was 0.64 +/- 0.10 (range, 0.50-0.77) with SH L 643A and 0.36 +/- 0.16 (range, 0.17-0.66) with gadopentetate dimeglumine (P <.01). The mean microvessel density (in terms of vessels per field of view) was 10.7 +/- 5.5 for postoperative scars and 30.0 +/- 7.7 for VX2 carcinoma (P <.001). CONCLUSION: The difference between the enhancement ratios of postoperative scars and VX2 carcinomas with SH L 643A was greater than that with gadopentetate dimeglumine. Enhancement ratios at SH L 643A-enhanced MR imaging corresponded well with microvessel density in postoperative scars and VX2 carcinomas. PMID- 14519874 TI - Osseous invasion by soft-tissue sarcoma: assessment with MR imaging. AB - PURPOSE: To assess magnetic resonance (MR) imaging signs and overall accuracy of MR imaging for detection of osseous invasion by soft-tissue sarcoma, with histopathologic correlation as the reference standard. MATERIALS AND METHODS: Preoperative MR images (1.5 T, transverse and longitudinal planes, T1 and T2 weighted) of 56 osseous sites in 51 patients who underwent bone resection at surgery for soft-tissue sarcoma were assessed retrospectively for signs of osseous invasion, by consensus of two readers who were blinded to clinical and histopathologic findings. Delay between MR imaging and surgery averaged 34 days (range, 2-112 days). MR signs assessed included osseous abutment by tumor, maximal diameter of osseous abutment, extent of circumferential abutment of long bones (<25%, 25%-50%, >50%), cortical destruction, and cortical and medullary signal intensity change on T1- and T2-weighted images. Imaging findings were correlated with histopathologic findings. Sensitivities, specificities, positive and negative predictive values (PPV, NPV), and P values were calculated. RESULTS: Eleven sites (20%) showed osseous invasion histologically (two, cortical; nine, both cortical and medullary). Tumor abutted bone at 44 lesion sites (sensitivity, 100%; specificity, 27%). Maximal diameter of osseous abutment and extent of circumferential abutment did not significantly affect osseous invasion (P =.09 and.11, respectively). On T1- and T2-weighted images, 13 lesion sites showed cortical signal intensity change (sensitivity, 100%; specificity, 96%) and 10 showed cortical destruction (sensitivity, 82%; specificity, 98%). Eleven sites showed decreased medullary T1 signal intensity (sensitivity, 100%; specificity, 96%), and 12 showed increased medullary T2 signal intensity (sensitivity, 100%; specificity, 94%). MR imaging overall had a sensitivity of 100%, specificity of 93%, PPV of 79%, and NPV of 100% for detection of osseous invasion on the basis of any finding of cortical destruction or cortical or medullary signal intensity change on T1- or T2-weighted images (P <.001). CONCLUSION: On T1- and T2-weighted MR images, findings of cortical and medullary signal intensity change and cortical destruction were sensitive and specific for detection of osseous invasion by soft-tissue sarcoma. PMID- 14519875 TI - Effect of varying CT section width on volumetric measurement of lung tumors and application of compensatory equations. AB - PURPOSE: To determine how volume measurements of simulated and clinical lung tumors at standard computed tomographic (CT) lung window and level settings vary with section width and to derive and apply compensatory equations. MATERIALS AND METHODS: Spherical simulated tumors of varying diameters were imaged with varying CT section widths, the images were displayed on a workstation, the cross sectional area of the tumor on each section was measured by using elliptical and perimeter methods, and the areas were integrated to compute tumor volume. The actual and measured tumor volumes for differing section widths and tumor diameters were compared, and compensatory equations were derived. The equations were applied to contemporaneous chest CT images obtained in patients with stage I lung cancer, and the difference between thick- and thin-section-derived volumes before and after application of the equations was determined. RESULTS: All simulated tumor volumes were overestimated 11%-278%; overestimation varied directly with section width and inversely with tumor diameter. With both measurement methods, mean thin-section volumes of clinical tumors in 55 patients were significantly smaller (P <.01) than mean thick-section volumes: Mean elliptical measurements were 15,025 mm3 (thin) and 18,037 mm3 (thick), with a 20.0% difference; mean perimeter measurements were 16,164 mm3 (thin) and 20,718 mm3 (thick), with a 22.2% difference. The thin-section-to-thick-section volume difference was larger for the smallest tumors. Thin-section volumes were smaller than thick-section volumes in 53 patients with the elliptical method and in 51 patients with the perimeter method. Applying the equations decreased the difference between thick- and thin-section volumes in 37 (67%) of the 55 patients with the elliptical method and in 41 (74%) patients with the perimeter method. The mean thin-section-to-thick-section volume difference became nonsignificant with the perimeter method but remained significant with the elliptical method. CONCLUSION: Measured lung tumor volumes vary significantly with varying CT section width; overestimation varies directly with section width and inversely with tumor size. Compensatory equations that are somewhat effective in reducing these effects can be derived. PMID- 14519876 TI - Multi-detector row CT versus coronary angiography: preoperative evaluation before totally endoscopic coronary artery bypass grafting. AB - PURPOSE: To assess multi-detector row spiral computed tomography (CT) for preoperative evaluation of patients undergoing totally endoscopic coronary artery bypass grafting and to correlate the data with coronary angiographic and intraoperative findings. MATERIALS AND METHODS: Thirty-six patients preoperatively underwent multi-detector row CT (4 x 1-mm collimation, pitch of 1.5, 500-msec rotation time, retrospective electrocardiographic gating, 1.25-mm effective section thickness) and coronary angiography. Assessment criteria for both techniques were visibility and cardiac course of coronary arteries, localization and degree of stenoses, composition of atherosclerotic plaques, and vascular diameter at anastomosis site. Site for distal bypass anastomosis was recommended. Results at multi-detector row CT were calculated relative to results at coronary angiography and surgery. RESULTS: Multi-detector row CT properly displayed 79.4% (154 of 194) of all surgical relevant coronary segments and 80.4% (434 of 540) of all coronary segments. For coronary angiography, ratios of 88.7% (172 of 194) and 94.6% (511 of 540), respectively, were observed. For detection of calcified plaques, multi-detector row CT results exceeded those at coronary angiography by a difference of 17% (18 of 18 [100%] compared with 15 of 18 [83%]). Hemodynamically relevant stenoses were identified with multi-detector row CT in 76% (42 of 55) of cases. Bridging of coronary segments through either myocardium (four of five) or epicardial fat (two of three) was better identified at multi-detector row CT than it was at coronary angiography (one of five compared with zero of three, respectively). At multi-detector row CT, 76% (28 of 37) of all distal bypass touchdown segments were identified, but at coronary angiography, only 70% (26 of 37) were identified. CONCLUSION: Multi-detector row CT provides extended information about coronary target site and therefore should be regarded as an ideal additive planning tool for complex minimally invasive procedures such as totally endoscopic coronary artery bypass grafting or minimally invasive direct coronary artery bypass grafting. PMID- 14519878 TI - Solid breast masses diagnosed as fibroadenoma at fine-needle aspiration biopsy: acceptable rates of growth at long-term follow-up. AB - PURPOSE: To determine what growth rate is acceptable before recommending histologic diagnosis of solid breast lesions diagnosed as fibroadenoma at fine needle aspiration biopsy (FNAB). MATERIALS AND METHODS: For 1,070 consecutive patients with breast lesions diagnosed as fibroadenoma at FNAB, three measurements of each mass were performed at the initial visit when FNAB was performed and at each follow-up ultrasonographic examination. Changes in volumes were calculated. At one or more visits, 194 masses showed an increase in volume. Nonfibroadenomas were excluded, and the data were used for comparison. Percentiles (90th and 95th) for percentage change in volume per month were used to determine acceptable changes in dimensions (specifically, greatest anteroposterior, parallel-to-skin, and perpendicular-to-skin dimensions). RESULTS: There were 567 interval measurements of 179 masses in 173 patients younger than 50 years and 50 measurements of 15 masses in 14 patients 50 years or older at the time of FNAB. The 95th percentile for percentage change in volume per month was approximately 16% for patients younger than 50 years; the 90th percentile was approximately 13% for patients 50 years or older. The 95th percentile mean change in dimension in a 6-month interval for those younger than 50 years was 20%; the 90th percentile change for those 50 years or older was also 20%. All excised masses with slower growth proved benign at histologic examination. CONCLUSION: Solid breast masses diagnosed as fibroadenomas at FNAB may be safely followed up if volume growth rate is less than 16% per month in those younger than 50 years and less than 13% per month in those 50 years or older. Acceptable mean change in dimension for a 6-month interval is 20% for all ages. PMID- 14519877 TI - Benign and malignant breast lesions: diagnosis with multiparametric MR imaging. AB - PURPOSE: To both develop and use a tissue signature method for the identification and classification of breast lesions and healthy breast tissue with magnetic resonance (MR) imaging. MATERIALS AND METHODS: Thirty-six patients underwent breast MR imaging (T1- and T2-weighted imaging and three-dimensional T1-weighted imaging with and without contrast material enhancement), followed by biopsy or mastectomy and histopathologic analysis. Tissue cluster analysis was performed by using the iterative self-organizing data technique to identify glandular, adipose, and lesion tissue signature vectors. Glandular and lesion tissue vectors were characterized by angular separation from the reference adipose tissue vector. Differences in angular separation of histologically proved benign and malignant lesion groups were evaluated with an independent t test. The usefulness of the angular separation model for distinguishing benign from malignant lesions was evaluated with nonparametric receiver operating characteristic curve analysis. RESULTS: The model enabled successful identification and characterization of breast lesion tissue clusters in all patients; 18 lesions were benign, and 18 were malignant. Angular separation +/- SD was 17.8 degrees +/ 6.1 degrees between adipose tissue and malignant lesions and 29.0 degrees +/- 11.2 degrees between adipose tissue and benign lesions. Angular separations of benign lesions and malignant lesions were significantly different (P =.002), with a specificity of 78% and sensitivity of 89% at a cutoff value of 21 degrees. Significant differences in angular separation from adipose tissue also were found between glandular tissue and lesion tissue (P <.001) and, in glandular tissue, between patients with benign lesions and those with malignant lesions (P =.04). The area under the receiver operating characteristic curve was 0.84. CONCLUSION: Multispectral analysis of conventional breast MR images based on the iterative self-organizing data model and on measurement of angular separation between tissue signature vectors may enable automated lesion identification and classification. PMID- 14519879 TI - The soft-tissue rim sign. PMID- 14519880 TI - In vivo proton MR three-dimensional T1rho mapping of human articular cartilage: initial experience. AB - The purpose of this study was to demonstrate the feasibility of computing three dimensional relaxation maps of spin-lattice relaxation time in the rotating frame (T1rho) from in vivo magnetic resonance (MR) images of the human patellofemoral joint. T1rho was measured by applying a three-dimensional gradient-echo pulse sequence in six healthy subjects and one symptomatic subject by using a 1.5-T MR imager and a 15-cm-diameter transmit-receive quadrature birdcage radiofrequency coil. Average T1rho measured in healthy patellar cartilage was 49.7 msec +/- 3.2 (mean +/- SD). Two-dimensional T1rho-weighted images were obtained with a fast spin-echo pulse sequence for comparison. There was good correlation between two dimensional and three-dimensional T1rho values for the six healthy subjects (R2 = 0.88, slope = 1.16). PMID- 14519881 TI - Dual-radionuclide simultaneous biliary and gastric scintigraphy to depict surgical treatment of bile reflux. AB - Biliary diversion procedures are performed during gastric surgery to decrease bile reflux. A 1-day dual-radionuclide examination was studied to determine its potential in the evaluation of the effectiveness of the Braun enteroenterostomy in reducing bile reflux and its effects on gastric emptying. Orally ingested gallium 67-labeled egg and intravenously administered technetium 99m diisopropyl imino-diacetic acid were imaged simultaneously. This provided a way to depict both bile reflux and gastric emptying on the same day in patients who underwent gastric surgery. Overall, the Braun enteroenterostomy trades bile reflux, a symptomatic and premalignant disease, for gastroparesis, a less severe and often treatable disease. PMID- 14519882 TI - Missed mammographic abnormalities, malpractice, and expert witnesses: does majority rule in the courtroom? PMID- 14519883 TI - Responsible use of CT. PMID- 14519884 TI - Urinary bladder pseudomass as a postoperative US finding. PMID- 14519885 TI - By-patient performance characteristics of CT colonography: importance of polyp size threshold data. PMID- 14519886 TI - Peptide-treated stent graft for the treatment of saphenous vein graft lesions: first clinical results. AB - Percutaneous interventions of degenerated saphenous vein grafts (SVG) are associated with high rates of distal embolic events and in-stent restenosis. A novel ePTFE-covered stent graft has been developed to improve outcomes for these difficult lesions. The stent graft has been treated with a 15-amino acid, cell adhesion peptide (P-15) to promote endothelialization on the inner surface post implantation. Patients with de novo lesions in SVGs were treated with a peptide coated ePTFE stent graft. Follow-up data was collected at 30 days, 3 and 6 months. An angiographic core lab analyzed an angiographic follow-up at 6 months. RESULTS: Procedural success was achieved in all cases. No device-related major adverse cardiac events occurred in-hospital, at 30 days, or at 3 months, despite the lack of distal protection. Six-month restenosis rates were 21%, and were comparable to similar devices. CONCLUSIONS: The peptide-treated stent graft appears to be a safe and effective treatment for SVG lesions, and has the potential to reduce complications without the use of adjunct distal protection in this patient population. PMID- 14519887 TI - Comparison of stainless steel stents coated with turbostratic carbon and uncoated stents for percutaneous coronary interventions. AB - BACKGROUND: Stent coating with turbostratic carbon was supposed to minimize the local inflammatory response after stent implantation and to thereby also reduce the rates of restenosis and clinical events. METHODS AND RESULTS: From October, 1999 to February, 2002, a total of 329 patients with symptomatic coronary artery disease (CAD) eligible for single-lesion PCI were randomized for implantation of either a CarboStent (C; n = 168) or a stainless-steel stent (S; n = 161). The stainless-steel stents were Tristar stents in 60.2%, Tetra stents in 17.4% and Penta stents in 22.4%. Both groups showed no differences in baseline clinical and angiographic characteristics. Angiographic follow-up (FU) was obtained after 6 months in 287 patients (87.2%), clinical FU in 295 patients (89.7%). With the exception of a smaller post-procedure minimal luminal diameter (MLD) in the C group (2.59 0.43 mm versus 2.72 0.46 mm in the S group; p = 0.01), there were no significant differences between the C and S groups in lesion length (10.28 4.45 mm versus 10.37 4.79 mm, respectively), reference diameter (2.92 0.59 mm versus 2.89 0.53 mm, respectively), pre-procedure MLD (0.77 0.36 mm versus 0.84 0.36 mm, respectively), MLD at FU (1.67 0.64 mm versus 1.68 0.57 mm, respectively), late loss (0.93 0.63 mm versus 1.05 0.59 mm, respectively), late loss index (0.51 0.32 versus 0.57 0.32, respectively) and restenosis rate (18.1% versus 20.6%, respectively). There were also no significant differences regarding major adverse cardiac events (MACE) between the C and S groups, i.e., occurrence of death (0% versus 0.7%, respectively), myocardial infarction (0% versus 0.7%, respectively), coronary artery bypass graft (0.7% versus 1.4%, respectively) and target lesion revascularization (16.4% versus 21.5%, respectively). CONCLUSION: Coronary stents coated with turbostratic carbon gave no clinically relevant reduction of in-stent restenosis and MACE rates when compared to uncoated stents. PMID- 14519888 TI - Passive coating: the dream does not come true. PMID- 14519889 TI - The influence of diabetes mellitus on the practice and outcomes of percutaneous coronary intervention in the community: a report from the HCA database. AB - BACKGROUND: Diabetics remain a high-risk group for those undergoing percutaneous coronary intervention (PCI) using balloon angioplasty and/or intracoronary stents for myocardial revascularization. The objective of this study is to compare clinical characteristics, demographics, procedure indications, lesion characteristics, and acute and long-term outcomes between diabetic patients and non-diabetic patients in a community based PCI registry. METHODS AND RESULTS: Information on patient demographics, coronary risk factors, lesion characteristics, procedures, and outcomes were derived from an HCA, Inc. database on all patients undergoing a PCI procedure in one of four community cardiac catheterization laboratories (CCL). A total of 3,139 patients who underwent PCI procedures from July 1, 1999 through September 30, 2000 were enrolled in this study. Approximately one-third of these patients completed a follow-up survey one year after their initial encounter. Analysis was limited to those patients undergoing PCI of native vessels with stents or conventional balloon angioplasty; patients with target lesions in bypass grafts or those treated with atherectomy were excluded. Approximately 23.5% of the patients enrolled in the study were diabetic. This study found no significant difference in any acute outcome between diabetic and non-diabetic patients in the hospital episode associated with the index PCI procedure. However, data from the 1-year follow-up survey indicates diabetic patients tended to have more target lesion revascularization (TLR) (13.6% versus 8.9%; p = 0.07) and more target vessel revascularization (TVR) (17.6% versus 12.7%; p = 0.058) than non-diabetic patients. In addition, adjusted odds ratios indicate that diabetic patients were 1.6 times more likely to have a second PCI procedure in another vessel (p = 0.013), 2.4 times more likely to undergo bypass surgery (p = 0.003), 1.9 times more likely to undergo an additional revascularization procedure (p < 0.001) and 1.8 times more likely to experience any major adverse cardiac events (p < 0.001) than non-diabetic patients during the follow-up period. CONCLUSIONS: This study indicates that selected diabetic patients can be treated for myocardial revascularization using PCI procedures with acceptable acute outcomes. However, diabetic patients undergoing PCIs have significantly more disease progression and are more likely to experience the need for recurrent revascularization. PMID- 14519890 TI - Diabetes mellitus and preferred method of coronary revascularization--the debate continues. PMID- 14519891 TI - Updated risk adjustment mortality model using the complete 1.1 dataset from the American College of Cardiology National Cardiovascular Data Registry (ACC-NCDR). AB - OBJECTIVES: To revise and update a risk adjustment model for in-hospital mortality following percutaneous coronary intervention (PCI) procedures using all data from the 1.1 version of the American College of Cardiology National Cardiovascular Data Registry (ACC-NCR). BACKGROUND: A model based on data received at the ACC-NCDR from 1998-2000 was previously reported. The revision of this mortality model reflects all of the data submitted using 1.1 data specifications and collected through the second quarter of 2001. The model was applied to selected high-risk subgroups from a sample of data collected during the year 2001 from version 2.0 of the NCDR. METHODS: Data on 173,743 PCI procedures collected at the ACC-NCDR between January 1, 1998 and March 31, 2001 were analyzed. A mortality model was generated as well as separate models for presentation with and without acute myocardial infarction within 24 hours. The model was used to generate predicted mortalities that were compared to observed mortalities in more current high-risk patient subgroups in the NCDR. RESULTS: The same factors that were previously found to be associated with increased risk of PCI mortality were re-verified in the current analysis. Inclusion of the complete 1.1 dataset produced some changes in the regression weights and the constant value. Excellent discrimination was achieved in the revised model (C-Index = 0.89). The model was applied to high-risk patient groups from data collected on 76,249 during the calendar year 2001 using the 2.0 NCDR data elements and definitions. These analyses showed a high level of agreement between observed mortality of each subgroup and the predicted mortality rates generated from the revised 1.1 PCI mortality model. CONCLUSIONS: Risk adjustment models for in hospital mortality following PCI for all patients and for those with and without recent MI were regenerated using all data collected from the 1.1 data specifications of the ACC-NCDR and validated on high-risk groups from data collected during 2001 under data version 2.0 of the NCDR. These models reflect the most up-to-date analysis of mortality prediction from this large, multi center national database. PMID- 14519892 TI - Pilot study of oral rapamycin to prevent restenosis in patients undergoing coronary stent therapy: Argentina Single-Center Study (ORAR Trial). AB - Rapamycin-coated stents are associated with low restenosis rates, but the ability of oral rapamycin to prevent restenosis is unknown. From December 2001 through February 2002, thirty-four patients with 49 lesions were treated with oral rapamycin for 1 month following percutaneous coronary intervention (PCI) with bare stents. Patients received a loading dose of 6 mg rapamycin followed by a daily dose of 2 mg. Rapamycin blood levels were measured in all patients during the third week of treatment. Cholesterol and triglycerides were evaluated before and 1 month after treatment. A 6-month follow-up angiogram was performed in all patients. Angiographic binary restenosis (> 50%), target lesion revascularization (TLR), late loss, treatment compliance and major adverse cardiac events were analyzed independent of rapamycin levels. Baseline characteristics included a history of diabetes in 35% of patients and the presence of in-stent restenosis in 24.5% of lesions (12/49). The rapamycin was well tolerated and only 1 patient discontinued the therapy due to mild side effects. Angiographic restenosis and TLR at 6 months was present in 26.5% of lesions (13/49). Restenosis in de novo lesions was 18.9% (7/37) compared to 50% of in-stent restenotic lesions (6/12; p = 0.08). Restenosis in de novo lesions in patients with rapamycin levels > 8 ng/ml was 0% (0/12), whereas it was 24% (6/25) when the rapamycin levels were < 8 ng/ml (p = 0.07). Late loss was significantly lower when rapamycin levels were > 8 ng/ml (0.3 mm versus 0.9 mm, respectively; p = 0.04). Thus, in this observational study, oral rapamycin administered for 1 month after PCI with bare stenting was safe and well tolerated. Higher therapeutic rapamycin blood levels were associated with a lower late loss and a trend toward a lower restenosis rate in de novo lesions. PMID- 14519893 TI - How should diuretic-refractory, volume-overloaded heart failure patients be managed? AB - Hospitalization is often required for patients with decompensated heart failure, usually with volume overload. Intravenous diuretics are recommended to optimize volume status; however, the management of patients refractory to diuretic therapy is not addressed in American Heart Association/American College of Cardiology, European Society of Cardiology or Heart Failure Society of America guidelines. This review focuses the invasive/interventional cardiologist on the rationale for greater involvement in the care of patients admitted to the hospital with decompensated volume overload, in particular when such patients are diuretic refractory. PMID- 14519894 TI - Catheter-based revascularization of the hepatic artery to treat coronary steal from a gastroepiploic artery bypass graft. AB - During diagnostic cardiac catheterization for symptoms of progressive angina, a high-grade ostial stenosis of an aberrant hepatic artery was identified. The common hepatic artery, which originated directly from the aorta rather than the celiac artery, supplied an in situ gastroepiploic artery bypass graft to the distal right coronary artery. Reduced flow within the bypass graft was observed consistent with a steal phenomenon from the coronary artery. This report describes successful angioplasty and stenting of the hepatic artery to salvage the gastroepiploic bypass graft and resolve the patient s ischemic symptoms. PMID- 14519895 TI - Left atrial thrombus formation on a CardioSeal Septal Occlusion device in a patient with elevated factor VIII: resolution with medical therapy. AB - We describe a case of thrombus formation on the left atrial disc of a CardioSeal Septal Occlusion device 14 days after transcatheter closure of a patent foramen ovale in a patient with a history of multiple cryptogenic strokes. Work-up to rule out the presence of a hypercoagulable state prior to device closure demonstrated an elevated factor VIII level. Medical therapy with heparin and aspirin was initiated, with subsequent clot resolution. PMID- 14519896 TI - Successful exclusion of descending thoracic aortic pseudoaneurysm by endovascular stent-graft placement. AB - Endograft Stenting for contained rupture in descending thoracic aorta has not been reported in the past. We successfully deployed a 28 mm long AneuRx cuff, percutaneously to exclude the descending thoracic aortic pseudoaneurysm in a 62 year-old patient with inoperable retroperitoneal malignant fibrous histiocytoma. PMID- 14519897 TI - Coronary fistula from left main stem to main pulmonary artery. AB - We describe a patient with a rare coronary arteriovenous fistula connecting the left main stem to the main pulmonary artery. This rare case was discovered during routine coronary angiography for the evaluation of the patient s coronary heart disease. PMID- 14519898 TI - PCI for totally occluded site consisting of huge thrombus. PMID- 14519899 TI - Milking-like effect secondary to systolic expansion of a post-infarction left ventricular aneurysm. AB - Systolic compression of coronary arteries is almost always due to the existence of myocardial bridging. We present a patient with a post-infarction left ventricular aneurysm in whom coronary angiography showed a milking-like effect with systolic compression of the distal left anterior descending coronary artery. PMID- 14519900 TI - Late termination of pregnancy. Professional dilemmas. AB - Abortion is an issue as long as history and hotly debated in all societies and communities. In some societies and countries it is legal, while other countries have no legal basis, and some countries have made it a crime. Today up to 90% of abortions take place in the first trimester, about 9% in the second trimester, and the rest in the third trimester. This paper deals with the issue of late termination of pregnancy, the practical medical aspects, legal issues, international aspects, and the dilemma for the professional. In early history, abortion was accepted by clergy and societies, but in recent history it is more restricted and in some countries prohibited. It does not seem that restriction leads to a lower abortion rate, but rather an active contraceptive policy, campaign, and availability to prevent pregnancies that are unwanted. In countries where abortion is restricted, the trend has been an increase in illegal abortion that leads to unsafe abortion with complications, permanent injuries, and maternal mortality. Unsafe and illegal abortion is a public health concern that governments should try to prevent and instead find ways to strengthen their commitments toward better and safer health and family planning services for women. Late termination of pregnancies is an issue of grave concern with many practical medical aspects, ethical questions, and professional dilemmas. This is especially of concern because of the viability of the fetus and should only take place in order to prevent harm to the physical and mental health of the mother or due to an anomaly or disability of the fetus. PMID- 14519901 TI - Evaluation of the most current and effective methods in the analysis of chlorinated dioxins in ground beef. AB - Chlorinated dioxins are the group of environmental pollutants consisting of 210 chlorinated dibenzo-p-dioxins and dibenzofurans. They are highly toxic and persistent. They are lipophilic and can easily biomagnify in the food chain, hence posing a serious threat to human health. The daily consumption of low-level contaminated food, mainly of animal origin, leads to the accumulation of dioxins in the human body. The exposures of the general human population to dioxins and the specific issues of a risk assessment of dioxin pose serious concerns in public environmental and nutritional health. This paper reviews the analysis of chlorinated dioxins in ground beef. The sources of contamination of chlorinated dioxins in ground beef are first reviewed to form a basis for a clear understanding of the health implications of chlorinated dioxins in the human food chain and why it is necessary to monitor the level of dioxins in animal food products, especially ground beef. The methods of collection, sampling, and processing of ground beef, and the methods of sample clean up prior to the analysis, are reviewed. Emphasis is laid on the new techniques that are available and that might be effective in the analysis of chlorinated dioxins in ground beef. Among these new methods and techniques are: the synergistic combination of ELISA/GC/MS, direct sample introduction to /GC/MS-MS, automated clean-up method, and the supercritical fluid extraction methods. The possible treatments of results from each method and technique are discussed and their respective efficiencies are compared. Finally, quality control and quality assurance parameters are evaluated for levels of accuracy, reproducibility, and precision. PMID- 14519902 TI - The Orthodontic Staff Shortage. PMID- 14519903 TI - An Essix intrusion appliance. PMID- 14519904 TI - Treatment of an adult patient with vertical maxillary excess using miniscrew fixation. PMID- 14519905 TI - Preventing accidents with demonstration casts. PMID- 14519906 TI - A method for eruption of impacted teeth. PMID- 14519907 TI - An extrusion spring for palatally impacted cuspids. PMID- 14519908 TI - The Inman Aligner. PMID- 14519909 TI - Human tibia bone marrow: defining a model for the study of haemodynamics as a function of age by near infrared spectroscopy. AB - A human model allowing the non-invasive study of bone marrow haemodynamics has been developed. A decrease in postischaemic tissue reperfusion capability (postischaemic hyperaemia) as a function of age (range 25-72 years) was observed both in the human tibia and tibialis anterior muscle. In the tibia bone marrow the reperfusion capability started to decrease after 50 years and was lower than for muscle for all the age range. Mean basal muscle O(2) saturation (80.8% at 25 years) decreases as a function of age (-0.35%+/-0.13% per year) whereas it remains constant for bone marrow (84.8+/-2.8%). A Monte Carlo simulation has been performed to evaluate the accuracy of the derived O(2) saturation measurements and has shown that this parameter is robust even in the presence of substantial noise. It has also been demonstrated that it is necessary to use a multi wavelength NIR spectrometer and a second derivative based fitting algorithm to obtain reliable measurements from the bone marrow, and that the tissue scattering changes occurring during the protocol do not allow the use of the standard near infrared spectroscopy algorithms. The human tibia bone marrow model presented here and the related measurement technique should enable access to new areas of physiological research. PMID- 14519910 TI - Physiological benefits of 24-style Taijiquan exercise in middle-aged women. AB - This study examined the physiological benefits of 24-style Taijiquan (24TJQ) exercises by comparing heart rate (HR), respiratory rate (RR), exercise intensity, electroencephalograph, surface electromyography and surface thermograph, as well as the results of physical fitness test in 20 middle-aged women (10 skilled participants and 10 novices). The data from the skilled participants showed greater values in sit-ups (p<0.01), side step (p<0.01) and stand trunk flexion (p<0.05), moreover, the statistic data demonstrated not only greater HR (p<0.05) or lower RR during exercise, but also higher beta%-power during the experiment, higher alpha%-power in the eye-closed period and central alpha dominant after exercise. These results suggest that 24TJQ is effective to promote physiological benefits in middle-aged women. It attracts strong interest and is helpful to induce psychological relaxation and mental concentration. PMID- 14519911 TI - Simulated high altitude diving experiment for the underwater construction operation. AB - The simulated dive experiments were conducted at the high altitude of 4500 meters and 5000 meters, for the requirement of diving operation in the lakes at the altitude of 4442 meters for the construction of large-scale hydroelectric power station. The high & low pressure chamber-complex was used, and 15 professional divers participated in the experiment. The divers were stayed at the altitude of 4500 and 5000 meters for 7-9 days. Totally 85 persons-times of dives to the depths of 30-50 meters were operated; they stayed under the water for 30-90 minutes while processing physical activities. During the experiment, we studied the pressurization procedure, decompression table, and physiological functions of the divers. The results indicate that, although the relative pressure differences between the surface and underwater was larger at high altitude than at sea level, the appropriate prolongation of the compression time was able to prevent the difficulty in pressure regulation for the divers to avoid the injury of middle ear. Four tables of the decompression A, B, C and D was calculated with Haldane's theory, and the speed of decompression increased in the order from A to D. The safest procedure was C, and there was no decompression sickness and bubbles in body of the divers. The methods of decompression included underwater stage decompression, surface decompression, oxygen-breathing decompression, and repetitive diving decompression. The surface decompression was the most suitable method for the high altitude, as it could greatly decrease the time in the cold water for the divers. The power spectrum analysis of EEG (electroencephalogram) indicated that, when the divers were exposed to the altitude of 5000 meters, the delta activity in EEG increased, alpha and beta activity decreased. And the delta activity decreased, the alpha and beta activity increased while diving during a dry condition. According to the diving and decompression procedure studied under simulated conditions, 272 person-times of diving training and underwater operations were processed in a high altitude hydroelectric power station at the altitude of 4442 meters, including photographing, video-recording, measuring, and drilling. There were no signs and symptoms of decompression sickness and bubbles. PMID- 14519912 TI - Ethnic differences in the relationship between bioelectrical impedance and body size. AB - The present study compared the regression equations of bioelectrical impedance on body size among various groups to investigate potential differences due to ethnicity. Data consisted of 30 Japanese and 28 Caucasoid subjects, and other groups of Aborigines, Danes, Melanesians and Polynesians from literature. The relationship between impedance and body weight fot the groups showed the ethnic difference. In the regression equations for Japanese and Caucasoid, a statistically significant difference was observed between both groups. The regression equation for Japanese was lower in the elevation. This seemed to be attributable to differences in the volume of fat-free mass for the same body build, configuration of the body, and fat-free mass density. PMID- 14519913 TI - An adult with atopic dermatitis and repeated short-term fasting. AB - It has been reported that nutritional stress, such as short-term fasting and long term energy restriction, has a suppressive effect on allergic dermatitis in experimental animals. Furthermore, clinical study has demonstrated a positive association between weight loss by low-energy diet and improvement in patients with atopic dermatitis. In this report, a 23-year-old female with atopic dermatitis received a treatment of repeated short-term fasting. 24-hour fasting was conducted once a week for a period of 20 weeks. On the fasting day, the amount of energy intake was 200 kcal. No medication was administered during the trial period. Clinical symptoms were evaluated using the Scoring Atopic Dermatitis index, and IgE, lactase dehydrogenase-5, and number of eosinophils were measured. At the end of the trial, body weight was reduced and clinical symptoms improved, whereas no improvements in laboratory findings were shown. For sufficient evidence of the effects of fasting, additional controlled study is needed. PMID- 14519914 TI - Formulation study for lansoprazole fast-disintegrating tablet. III. Design of rapidly disintegrating tablets. AB - Lansoprazole fast-disintegrating tablets (LFDT) are a patient-friendly formulation that rapidly disintegrates in the mouth. LFDT consist of enteric coated microgranules (mean particle size, approximately 300 microm) and inactive granules. In the design of the inactive granules, mannitol was used as a basic excipient. Microcrystalline cellulose, low-substituted hydroxypropyl cellulose (L HPC), and crospovidone were used as binders and disintegrants. A new grade of L HPC (L-HPC-33), with a hydroxypropoxy group content of 5.0-6.9%, was developed and it has no rough texture due to a decrease in water absorption. It was clarified that L-HPC-33 could be useful as a binder and disintegrant in rapidly disintegrating tablets. LFDT contain enteric-coated microgranules in tablet form. The enteric-coated microgranule content in LFDT affect qualities such as tensile strength, disintegration time in the mouth, and dissolution behavior in the acid stage and in the buffer stage of LFDT. The 47.4% content of the enteric-coated microgranules was selected to give sufficient tensile strength (not less than 30 N/cm(2)), rapid disintegration time in the mouth (not more than 30 s), and dissolution behavior in the acid stage and buffer stage similar to current lansoprazole capsules. Compression force affected the tensile strength and the disintegration time in the mouth, but did not affect the dissolution behavior in the acid and buffer stages. PMID- 14519915 TI - Reactions of indole derivatives with cardioprotective activity with reactive oxygen species. Comparison with melatonin. AB - We have previously reported on the synthesis of novel indole derivatives containing an amine-triazole moiety (1a-d, 2a-c), and their antioxidant activity on in vitro non-enzymatic rat hepatic microsomal lipid peroxidation. Some of the compounds showed protective activity against oxidative injury of ischemic myocardium. In the present paper we investigated the interactions of these derivatives with reactive oxygen species, in order to find a mechanism of their antioxidant capacity and to identify structural characteristics responsible for these properties. These interactions were compared with melatonin, which is also an indole derivative. The antioxidant profiles of the compounds were established by different in vitro protocols as follows: 1) by the interaction of the compounds with the 1,1-diphenyl-2-picrylhydrazyl (DPPH) stable free radical, 2) their scavenging effects on superoxide anions using an enzymic system of xanthine xanthine oxidase, 3) their inhibitory effects on xanthine oxidase and 4) their ability to scavenge hydroxyl radicals by comparison with dimethyl sulfoxide (DMSO) for *OH. All compounds were found to interact with DPPH, most of them to be superoxide anion scavengers and to be strong hydroxyl radical scavengers. Derivatives 1a and 1d substituted on the nitrogen of the indolic nucleus were found to have better antioxidant properties than the reference compounds used and melatonin. PMID- 14519916 TI - Molecular interactions of new pregnenedione derivatives. AB - The in vitro inhibitory activity of five new progesterone derivatives: 17alpha hydroxy-16beta-methylpregna-1,4,6-triene-3,20-dione 1; 16beta-methyl-17alpha toluoyloxypregna-1,4,6-triene-3,20-dione 2; 17alpha-hydroxy-6-methylenepregn-4 ene-3,20-dione 3; 6-methylene-17alpha-toluoyloxypregn-4ene-3,20-dione 4 and 17alpha-(p-bromobenzoyloxy)-6-methylenepregn-4-ene-3,20-dione 5 was determined. These compounds were evaluated as 5alpha-reductase inhibitors as well as antagonists for the androgen receptor. Compounds 1, 2, 3, 4 and 5 showed the following inhibitory activity for the 5alpha-reductase enzyme with IC(50) values of: 1 (1.65 microM), 2 (10 microM), 3 (19 nM), 4 (100 nM) and 5 (100 nM). The results of this study also showed the effect of increasing concentrations of the novel steroids upon [(3)H]dihydrotestosterone binding to androgen receptors from male hamster prostate. The K(i) values for compounds 1, 2, 3, 4, 5 and dihydrotestosterone showed the following order of affinity for the androgen receptor: 4>5>dihydrotestosterone>2>3>1. The overall data indicated that all synthesized compounds 1, 2, 3, 4 and 5 are inhibitors of the 5alpha-reductase enzyme present in the hamster prostate. In addition compounds 1, 2, 3, 4 and 5 also presented an affinity for the androgen receptor. PMID- 14519917 TI - Facile one pot microwave induced solvent-free synthesis and antifungal, antitubercular screening of spiro [1,5]-benzothiazepin-2,3'[3'H]indol-2[1'H] ones. AB - Microwave activation coupled with dry media technique as a green chemistry procedure has been applied to synthesis of a series of some new title compounds. They have been obtained by the reaction of in situ synthesized 1,3-dihydro-3-[2 (phenyl/4-fluorophenyl)-2-oxoethylidene)-indol-2(1H)-one (4a, b) with substituted aminobenzenethiols (5a-d). The key intermediates 4a, b were also prepared in one step by this improved technique by reacting isatin and substituted acetophenones (2a, b). The results obtained under microwave irradiation when compared with that following conventional method demonstrate the versatility of the process. The title compounds 7a-e have also been screened for their antifungal and antitubercular activity, 7a and 7e showing maximum inhibition of growth of Alternaria alternata and Fusarium oxysporium and 7b, c, e revealing significant antitubercular activity. PMID- 14519918 TI - Three diterpenoids (excoecarins V1-V3) and a flavanone glycoside from the fresh stem of Excoecaria agallocha. AB - Three new diterpenoids, excoecarins V1-V3 (1-3) and a new flavanone glycoside (7) were isolated from the fresh stem of Excoecaria agallocha L. Their structures were elucidated as: 2alpha,3alpha,18-trihydroxy-3beta,20-epoxybeyer-15-ene (1), ent-2,3-secokaur-16-en-2,3-dioic acid (2), ent-3,4-seco-16alpha-hydroxyatis-4(19) en-3-oic acid (3), and 3,5,7,3',5'-pentahydroxy-2R,3R-flavanonol 3-O-alpha-L rhamnopyranoside (7) on the basis of spectroscopic data, chemical evidence, and/or X-ray analysis. PMID- 14519919 TI - Field survey of Glycyrrhiza plants in central Asia (2). Characterization of phenolics and their variation in the leaves of Glycyrrhiza plants collected in Kazakhstan. AB - A new prenylated flavanone, licoleafol, and a new prenylated dihydrostilbene, uralstilbene, together with four known compounds, 8-dimethylallyleriodictyol, sophoraflavanone B, gancaonin R, and 6-dimethylallyleriodictyol, were isolated from the leaves of Glycyrrhiza uralensis collected in Kazakhstan. HPLC analysis of the leaves of Glycyrrhiza plants collected in Kazakhstan showed that both G. uralensis-specific and Glycyrrhiza glabra-specific compounds were detected in the leaves of the morphologically intermediate-type plants, suggesting that the intermediate-type plant is a hybrid of G. glabra and G. uralensis. In addition, HPLC profiles of leaf extracts from offspring of intermediate-type plants were divided into the three types: the G. uralensis type, G. glabra type, and the intermediate type. From these results, it appears likely that the intermediate type plant back-crosses with G. glabra and G. uralensis to generate a G. glabra type plant and a G. uralensis-type plant, respectively. PMID- 14519920 TI - An efficient procedure for the preparation of (1S,3R)- and (1S,3S)-1-amino-3 (hydroxymethyl)cyclopentanes. AB - Enantiomerically pure (1S,3S)- and (1S,3R)-1-amino-3-(hydroxymethyl)cyclopentanes have been efficiently synthesized from L-aspartic acid. The title compounds are isosteres of ribose and may be used to construct nucleoside analogs with important antiviral and antineoplastic activities as demonstrated by a concise total synthesis of (+)-4'-deoxycarbapentostatin nucleoside. PMID- 14519921 TI - Conformation of 2-aminomethylpyrrolidine and 2-aminomethylpiperidine in ternary platinum(II) complexes: regulation of conformation of the diamine by the coexisting ligand. AB - Square-planar complexes with the formula [Pt(L(2))(L(1))](X)(2) x nH(2)O, where L(1) is S-2-aminomethylpyrrolidine (S-pyrda) or 2-aminomethylpiperidine (pipda) and L(2) is diammine (X=Cl), cyclobutane-1,1-dicarboxylato (cbdca) (X=none), 2,2' bipyridine (bpy) (X=NO(3)), or 1,10-phenanthroline (phen) (X=Cl), were prepared and the nature of the coordination of L(1) was examined by (1)H-NMR spectroscopy and X-ray crystallography. These 2-aminomethylazacycloalkane derivatives form five-membered chelate rings condensed with an azacycloalkane ring in cis- or trans-configurations. The (1)H-NMR spectrum of complexes with S-pyrda as L(1) were consistent with cis-condensed rings in an S(N) conformation with any of L(2) group. However, (1)H-NMR spectra of the complexes with pipda as L(1) indicated trans-fused successive rings for the diammine and cbdca as L(2), but spectra for bpy and phen as L(2) were consistent with a conformation having cis-fused successive rings. X-Ray crystallography data for the two complexes with pipda as L(1) and cbdca (1) and bpy (2) as L(2) confirms the different coordination behavior in the solid state. PMID- 14519922 TI - Ten new Lycopodium alkaloids having the lycopodane skeleton isolated from Lycopodium serratum Thunb. AB - Ten new alkaloids, lycoposerramines-F (1), -G (2), -H (3), -I (4), -J (5), -K (6), -L (7), -M (8), -N (9), and -O (10), having lycopodine-related structures, were isolated from the club moss Lycopodium serratum THUNB. and their structures were elucidated on the basis of spectroscopic analysis and/or chemical transformation. PMID- 14519923 TI - Convenient indole synthesis from 2-iodoanilines and terminal alkynes by the sequential Sonogashira reaction and the cyclization reaction promoted by tetrabutylammonium fluoride (TBAF). AB - The sequential Sonogashira reaction and the cyclization reaction of various 2 iodoanilines and terminal alkynes in the presence of a palladium catalyst and tetrabutylammonium fluoride (TBAF) gave the corresponding 2-substituted indoles in good yields. PMID- 14519924 TI - Micranoic acids A and B: two new octanortriterpenoids from Schisandra micrantha. AB - Two novel octanortriterpenoids, micranoic acids A (1) and B (2), along with three known compounds, kadsuric acid (3), 3beta-hydroxy-lanost-9(11),24(25)-dien-26-oic acid (4) and schizandronic acid (5), were isolated from the leaves and stems of Schisandra micrantha. The structures of 1 and 2 were determined by 1D and 2D NMR spectroscopic analysis. Micranoic acids A and B represent a new group of triterpenes in which the entire C-17 side chain has lost. This is the first report of octanortriterpenoids isolated from the family Schisandraceae. PMID- 14519925 TI - Formal synthesis of squalamine from desmosterol. AB - The key intermediate to squalamine, (5alpha,7alpha,24R)-7,24-dihydroxy-cholestan 3-one, was synthesized from the 3-O-acetyl-24R,25-dihydroxy derivative of desmosterol via 10 steps in 16% overall yield and squalamine was also prepared via two further steps in 7.4% total yield from the desmosterol derivative. PMID- 14519926 TI - Enhanced skin permeation of cationic drug ketotifen through excised guinea pig dorsal skin by surfactants with different electric charges. AB - Using excised guinea pig dorsal skin, we examined the effects of three surfactants, anionic sodium dodecylsulfate (SDS), cationic n dodecyltrimethylammonium bromide and non-ionic n-dodecyl-beta-D-maltoside, all of which commonly have an n-dodecyl group, on in vitro skin permeation of the cationic drug ketotifen. All these three surfactants increased the skin permeation of ketotifen. Among the surfactants tested, anionic SDS had the largest enhancement effects, and significantly increased the permeation at concentrations over 1 mM. The enhancement effect of the same anionic surfactant on the permeation of anionic salicylate was smaller and similar to that of cationic n-dodecyltrimethylammonium. The enhancement effects of SDS on ketotifen permeation were more marked than those of the cationic surfactant but differed from previous findings of their effects on other drugs permeation. Analysis of the retention of ketotifen in the skin suggested that SDS-induced increase in the transfer of hydrophilic ketotifen to the skin is the main reason for the marked increase in skin permeation. PMID- 14519927 TI - A new flavone C-glycoside from the style of Zea mays L. with glycation inhibitory activity. AB - A new flavone C-glycoside, chrysoeriol 6-C-beta-fucopyranoside (1), and a known flavone C-glycoside (2) were isolated from the style of Zea mays L. These structures were elucidated on the basis of spectroscopic analysis. Compound 1 exhibited glycation inhibitory activity. PMID- 14519928 TI - New ent-kaurene-type diterpenoids possessing cytotoxicity from the New Zealand liverwort Jungermannia species. AB - Two new ent-kaurene-type and a new rearranged ent-kaurene-type diterpenoids possessing cytotoxicity against a human leukemia cell line have been isolated from the New Zealand liverwort Jungermannia species, together with previously known ent-kaurene-type diterpenoids. Their structures were established based on extensive NMR techniques. PMID- 14519929 TI - New sphingolipids from marine sponge Iotrochota baculifera. AB - Chemical examination of the marine sponge Iotrochota baculifera of the Indian Ocean furnished a sphingolipid (1), a new glycosphingolipid (iotroridoside-B, 2) and a four-component mixture of sphingolipids (3a-d) with two new components. Structure elucidation was carried out by spectral and chemical methods. PMID- 14519930 TI - Hydrophobicity parameters determined by reversed-phase liquid chromatography. XVI: A new hydrogen-accepting parameter for monosubstituted thiophenes and furans for correlating retention factors and octanol-water partition coefficients. AB - We recently proposed a new hydrogen-accepting parameter, S(HA), for monosubstituted (di)azines on the basis of the heat of formation calculated by the conductor-like screening model (COSMO) method. In this work, S(HA) values for monosubstituted thiophenes and furans were calculated and the results were applied to the analysis of relationships between log P (P: 1-octanol/water partition coefficient) and log k (k: retention factor obtained by reversed phase HPLC). The S(HA) parameter was found to work effectively as a hydrogen-bonding parameter in a range of heteroaromatic compounds. PMID- 14519931 TI - New aliphatic alcohol and (Z)-4-coumaric acid glycosides from Acanthus ilicifolius. AB - From the aerial parts of Acanthus ilicifolius, a new aliphatic alcohol glycoside (ilicifolioside C) and two new (Z)-4-coumaric acid glycosides, (Z)-4-coumaric acid 4-O-beta-D-glucopyranoside and (Z)-4-coumaric acid 4-O-beta-D-apiofuranosyl (1"-->2')-O-beta-D-glucopyranoside were isolated. The structural elucidations were based on the analyses of spectroscopic data. Z-Form 4-coumaric acid glycosides were first isolated from plant. PMID- 14519932 TI - Flavone C-glycosides from Viola yedoensis MAKINO. AB - A new flavone C-glycoside, apigenin 6-C-alpha-L-arabinopyranosyl-8-C-beta-L arabinopyranoside, has been isolated from Viola yedoensis together with the known compounds, apigenin 6,8-di-C-alpha-L-arabinopyranoside, apigenin 6-C-alpha-L arabinopyranosyl-8-C-beta-D-glucopyranoside (isoschaftoside), apigenin 6-C-beta-D glucopyranosyl-8-C-alpha-L-arabinopyranoside (schaftoside), apigenin 6-C-beta-D glucopyranosyl-8-C-beta-L-arabinopyranoside (neoschaftoside), apigenin 6,8-di-C beta-D-glucopyranoside (vicenin-2), apigenin 6-C-alpha-L-arabinopyranosyl-8-C beta-D-xylopyranoside, apigenin 6-C-beta-D-xylopyranosyl-8-C-alpha-L arabinopyranoside, luteolin 6-C-beta-D-glucopyranoside (isoorientin) and luteolin 6-C-alpha-L-arabinopyranosyl-8-C-beta-D-glucopyranoside (isocarlinoside). The structures were determined by spectroscopic methods and new or revised (1)H- and (13)C-NMR spectral assignments are proposed for some compounds. PMID- 14519933 TI - A new furanoid diterpene from Caesalpinia pulcherrima. AB - A new cassane-type diterpene isovouacapenol E (1) was isolated from the leaves of Caesalpinia pulcherrima, together with the known compounds caesaldekarin A (3), spathulenol (4), caryophyllene oxide (5), phytol, and sitosterol. The structure of 1 was elucidated by spectral data interpretation. PMID- 14519934 TI - New oxindole alkaloids and iridoid from Carolina jasmine (Gelsemium sempervirens Ait. f.). AB - Three new gelsedine-type oxindole alkaloids, GS-1, GS-2, and GS-3, and one new iridoid, GSIR-1, were isolated from the stems and leaves of cultivated Carolina jasmine (Gelsemium sempervirens AIT. f.) and their structures were determined by spectroscopic analysis. PMID- 14519935 TI - Two new 15,16-seco-cycloartane glycosides from Cimicifuga rhizome. AB - Two new 15,16-seco-cycloartane glycosides (1, 2) were isolated from Cimicifuga Rhizome. Their structures were determined by spectroscopic analysis. PMID- 14519936 TI - A new application of sequence Fourier analysis to specific ligand-protein interactions. AB - A frequency (or sequence)-dependent rule seems to be conserved in a simple and concerted interaction of various human proteins. PMID- 14519937 TI - A facile method for deprotection of O-allylphenols. AB - Allyl aryl ethers can be easily cleaved by the use of 10% Pd/C under the mild and basic conditions. PMID- 14519938 TI - Inhibitory effects of nardostachin on nitric oxide, prostaglandin E2, and tumor necrosis factor-alpha production in lipopolysaccharide activated macrophages. AB - Nardostachin, which is an iridoid isolated from Patrinia saniculaefolia, was examined by assessing its effect on the production of tumor necrosis factor-alpha (TNF-alpha) and expression of 2 enzymes, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), in lipopolysaccharide (LPS)-stimulated Raw264.7 macrophages. This compound consistently inhibited the production of nitric oxide (NO) and TNF-alpha production in a dose-dependent manner, with respective IC(50) values of 12.3 and 16.2 microM. The decrease in quantity of NO products was accompanied by a decrease in the iNOS protein level, as assessed by Western blotting probed with specific anti-iNOS antibodies. In addition, this compound also reduced the COX-2 protein expression level and the attendant PGE(2) production in LPS-stimulated macrophages. These results suggest that nardostachin may be useful for inhibiting the production of inflammatory mediators such as TNF alpha, NO and PGE(2) in inflammatory diseases. PMID- 14519939 TI - Specific inhibition of hypoxia-inducible factor (HIF)-1 alpha activation and of vascular endothelial growth factor (VEGF) production by flavonoids. AB - Screening using a reporter under the control of the hypoxia-response element (HRE) identified several flavonoids and homoisoflavonoids that inhibit the activation of HRE under hypoxic conditions. Among various compounds, isorhamnetin, luteolin, quercetin, and methyl ophiopogonanone B (MOB) were effective at 3 to 9 microg/ml in inhibiting the reporter activity. The expression of vascular endothelial growth factor (VEGF) mRNA during hypoxia was also inhibited by MOB in HepG2 cells, but the effective doses were 10 to 20 microg/ml. MOB caused destabilization of hypoxia-inducible factor (HIF)-1alpha, as revealed by Western blotting, that was dependent on proteasome activity and the tumor suppressor, p53. The tubular formation and migration of human umbilical vein endothelial cells was also inhibited by MOB. MOB is expected to act as an inhibitor of angiogenesis. PMID- 14519940 TI - Effect of Bak Foong pills on exocrine pancreatic-bile secretion. AB - We have recently demonstrated that Bak Foong Pills (BFP), a well-known Chinese medicine widely used for treating gynecological disorders, stimulates human colonic epithelial anion secretion, which was mediated by intracellular cAMP and Ca(2+). The present study further investigated the effect of BFP on exocrine pancreatic-bile secretion using in vivo and in vitro approaches. Duodenal infusion of BFP ethanol extract (1 g/kg) in rats produced increases in the volume and protein output of pancreatic-bile juice, but did not affect its pH. Surgical ablation of vagal neural pathway slightly reduced the effect of BFP on the protein output and volume, indicating that the vagal nerve pathway was not the major player in medicating the effect of BFP on exocrine pancreatic-bile secretion. Using CAPAN-1 cell line, a human pancreatic duct cell line, in conjunction with the short-circuit current (I(SC)) measurements, we further demonstrated that BFP could directly stimulate pancreatic HCO(3)(-) secretion. Basolateral addition of BFP (600 microg/ml) produced averaged charges transported of 2100+/-382.5 microC/cm(2), which was blocked by apical addition of Cl(-) channel blocker. Removal of HCO(3)(-) from the Krebs-Henseleit (K-H) solution inhibited the BFP-induced I(SC) by more than 95%. The present results suggest that BFP could improve digestive function by stimulating pancreatic protein and HCO(3)(-) secretion. PMID- 14519941 TI - Effects of proteins from hen egg yolk on human platelet aggregation and blood coagulation. AB - We studied the effect on both platelet aggregation and blood coagulation, known to be major risk factors in thrombogenesis, of proteins from hen egg yolk (EP). EP potently inhibited collagen-induced human platelet aggregation in a dose dependent manner. Furthermore, EP has a synergistic effect on the inhibition of human platelet aggregation with both molsidomine, an inhibitor of cGMP-specific phosphodiesterase, and theophylline, an inhibitor of cAMP-specific phosphodiesterase. These results indicate that the active mode of EP might be involved in elevation of the levels of both cGMP and cAMP. Prothrombin time and activated partial thromboplastin time were potently prolonged by EP. These data suggest that EP prolongs the time interval between the conversion of fibrinogen to fibrin. Accordingly, these findings demonstrate that EP might have antithrombotic effects by inhibiting platelet aggregation and fibrin formation. PMID- 14519942 TI - Influence of histamine in a liver injury model induced by Propionibacterium acnes and lipopolysaccharide. AB - In normal mice, plasma histamine levels were 29.4+/-10.1 pmol/ml. When 0.1 microg of lipopolysaccharide (LPS) was intravenously injected into Propionibacterium acnes (P. acnes)-primed ICR mice, histamine levels increased remarkably to 61.2+/ 15.9 pmol/ml (p<0.001). An increase was also observed in liver tissues. Oral administration of histidine at 200 mg/kg once daily for 5 d before intravenous LPS injection increased the plasma alanine aminotransferase (ALT) activity to 2936.5+/-356.3 IU/l, a significant change compared with the controls (2244.8+/ 425.5 IU/l, p<0.05). The 24 h survival rate after LPS injection was 72.7% in the mice treated with 50 mg/kg of ranitidine, in contrast with 50% in the control group although the treatment did not significantly decrease the plasma ALT activity. On the other hand, 50 mg/kg of pyrilamine significantly reduced plasma ALT activity (p<0.001). The results suggested that histamine levels are related to hepatic damage in the P. acnes plus LPS induction of liver injury. PMID- 14519943 TI - Quercetin, a flavonoid antioxidant, prevents and protects against ethanol-induced oxidative stress in mouse liver. AB - This study evaluates whether quercetin (25, 50 and 75 mg/kg body weight) treatment has a protective effect on the pro-oxidant-antioxidant state following chronic ethanol treatment in mice. Pretreatment (quercetin 25, 50 and 75 mg/kg body weight for 15 d+co-treatment of ethanol 18%+quercetin for 15 d and ethanol 18% for the 15 d) increased the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione (GSH) in comparison to the ethanol group. No significant differences from the ethanol group were observed in the group after post-treatment (ethanol 18% for 30 d+quercetin 25, 50 and 75 mg/kg body weight for 15 d) with quercetin. A significant increase in lipid peroxidation (malondialdehyde, MDA) products was observed in liver tissue after administration of ethanol, which was attenuated by pre- and post-treatment with a high dose of quercetin. GSH levels increased and oxidized glutathione (GSSG) levels decreased in groups of ethanol-exposed mice that received quercetin for 15 d prior to ethanol exposure. In conclusion, pre treatment of quercetin may protect against ethanol-induced oxidative stress by directly quenching lipid peroxides and indirectly by enhancing the production of the endogenous antioxidant GSH. There was no protective effect on post-treatment with quercetin. PMID- 14519944 TI - Effect of the spinal drug tramadol on the fatty acid compositions of rabbit spinal cord and brain. AB - Tramadol is an important spinal drug which produces analgesia following intrathecal injection. It is well known that fatty acids (FAs) play an important role in membrane fluidity of the blood-brain barrier (BBB) tissue, which blocks and/or controls the transportation of toxic substances into the brain. The aim of this study was to investigate the effect of a spinal drug (tramadol) on the concentrations and compositions of fatty acid in BBB tissues of New Zealand male rabbits. The total cellular fatty acid profiles of the tissues in three spinal cord sections (cervical, thoracal and lumbar) and in the brain of rabbits with or without drug administration were determined by gas chromatography using Sherlock Microbial Identification System (MIS) software (Microbial ID, Newark, DE, U.S.A.) with a database of FAME profiles for eukary. The relative percentage of the fatty acid methyl ester (FAME), 24 : 1 omega9c nervonic and 17 : 1 omega8c, did not change with tramadol treatments. However, there was an increase in the concentration of the FA 16 : 0, 18 : 1 omega7c DMA, 18 : 1 omega9c, sum in future 4, sum in future 8, sum in future 9, 18 : 0, 20 : 4 omega6c, sum in future 14, 22 : 4 omega6c, in contrast to a decrease in the percentages of the following FAMEs; 20 : 0, 20 : 1 omega9c. In the brain, there was an increase in the concentration of the FA 18 : 1 omega9c, sum in future 8 and 18 : 0, in contrast to a decrease in the percentages of two FAMEs, 16 : 0, 20 : 4 omega6c and 22 : 6 omega3c. The number of fatty acids were 20 in the spinal cord sections and 8 in the brain tissues of control animals compared to 15-18 fatty acids in the spinal cord section and 7 in the brain tissues of drug administered animals. The overall changes in the concentrations and numbers of FAs suggest that the spinal drug tested in this study has a side effect of disrupting of membrane fluidity of the BBB, which may cause neurotoxicity. PMID- 14519945 TI - Synthesis and anticonvulsant activity of novel 1-substituted-1,2-dihydro pyridazine-3,6-diones. AB - The synthesis and pharmacological evaluation of novel 1-substituted-1,2-dihydro pyridazine-3,6-diones (4a--l, 5a--j) as potential anticonvulsant agents are described. The compounds were tested in vivo for the anticonvulsant activity. The compound which have maximum protection against MES induced seizures is 1-[3-(2 aminophenylamino)-2-hydroxypropyl)-1,2-dihydro-pyridazine-3,6-dione 4h (ED(50)=44.7 mg x kg(-1) i.p.) 1-[2-hydroxy-3-piperazin1-yl-propyl)-1,2-dihydro pyridazine-3,6-dione 4c (ED(50)=72 mg x kg(-1) i.p.) and 1-[2-hydroxy-3-imidazol 1-yl-propyl)-1,2-dihydro-pyridazine-3,6-dione 4d (ED(50)=79 mg x kg(-1) i.p.) were also found to have maximum protection against MES induced seizures. Whereas all these compounds failed to protect the animals from subcutaneous pentylenetetrazole (Metrozol) seizure threshold test (sc-Met). PMID- 14519946 TI - Potent cytotoxic effects of novel retinamide derivatives in ovarian cancer cells. AB - 4-(N-Hydroxyphenyl)retinamide (also known as 4-HPR or fenretinide), a synthetic amide of all-trans retinoic acid (RA), has been implicated as a promising anticancer agent associated with reducing the toxicity related to RA. However, the low plasma levels of 4-HPR in patients limited clinical trials, leading to a search for derivatives with better efficacy. In this study, we synthesized a series of 4-HPR derivatives in good yields by introducing acetate (compound 1). propionate (2). pyruvate (3). butyrate (4). or stearate (5). to the 4 hydroxylphenyl moiety of 4-HPR. In our initial proliferation assays, we identified compound 3 as the most cytotoxic of the series against four ovarian cancer cell lines (OVCAR-3, PA-1, 2774, and SKOV-3). Dose-response curves yielded IC(50) values of 3.75-7.75 microM for AtRA, 2.80-5.50 microM for 9-cis RA, 0.65 4.05 microM for 4-HPR, and 0.25-0.75 microM for compound 3, depending on the cell type treated. Nuclear staining with 4',6-diamidino-2-phenylindole (DAPI) and DNA fragmentation assays clearly indicated that the antiproliferative effect of compound 3 was mediated by apoptosis. In contrast to natural retinoids, both 4 HPR and compound 3 activated two (RARbeta and RARgamma) of the three retinoic acid receptor (RAR) subtypes tested, but did not activate any of the three retinoid X receptors (RXRs), as determined by transcription assays in OVCAR-3 cells. However, like natural retinoids, 4-HPR and compound 3 actively suppressed c-Jun transcriptional activity. Thus, compound 3 not only showed more potent antiproliferative activity than any other retinoid derivatives tested, but also effectively inhibited the c-Jun activity that has been implicated in tumor promotion and invasion. These results, together with compound 3's selectivity for RAR subtypes, suggest that compound 3 could be an effective anticancer drug for ovarian cancer, with less toxicity than RA. PMID- 14519947 TI - Alleviation of experimental septic shock in mice by acidic polysaccharide isolated from the medicinal mushroom Phellinus linteus. AB - This study reports that acidic polysaccharide (PL) isolated from Phellinus linteus alleviated the septic shock induced by high dose lipopolysaccharide (LPS) injection in mice. To examine the origin of this effect, we investigated cytokine production in serum and the expression of MHC II in B cells and macrophages in areas of inflammation. Pretreatment with PL 24 h before LPS administration resulted in a significant inhibition of up to 68% of circulating tumor necrosis factor (TNF)-alpha, a moderate reduction of 45% of interleukine (IL)-12 and 23% of IL-1beta, but no significant reduction in IL-6. In addition, the expression of MHC II in B cells and macrophages was examined. Our results show that LPS stimulated cytokine release and the level of MHC II can be modulated by in vivo administration of soluble PL in mice. The decrease of IL-1beta, IL-12 and TNF alpha in sera and the down-modulation of MHC II during septic shock may contribute to the long survival of mice by PL. Administration of PL in vivo decreases IL-2, IFN-gamma and TNF-alpha production in splencotyes and enhances spontaneous cell apoptosis in macrophages and lymphocytes stimulated with LPS in vitro. Thus, part of the anti-inflammatory effects of PL treatment in vivo may result from the enhanced apoptosis of a portion of the activated macrophages and lymphocytes. The ability of PL to significantly reduce the TNF-alpha production indicates the potential of the polysaccharides in possible therapeutic strategies that are based on down-regulation of TNF-alpha. PMID- 14519948 TI - Studies on the antioxidant activities of Desmodium gangeticum. AB - Desmodium gangeticum is herbal species which is widely used in the indigenous system of medicine and is reported to contain flavone and isoflavanoid glycosides. In view of its wide use and it's chemical composition, this study was aimed at examining the antioxidant activity of the extract of D. gangeticum. The extract was studied for diphenyl picryl hydrazyl (DPPH), nitric oxide, ferryl bipyridyl and hypochlorous acid scavenging activity along with lipid peroxidation. Nitric oxide was generated using sodium nitroprusside and was studied using Griess reagent. In order to study the iron chelating capacity of the extract, the percentage ferryl-bipyridyl inhibition was studied. Hypochlorous acid scavenging activity was tested by measuring the inhibition of 5-thio-2 nitrobenzoic acid oxidation. The extract was also studied for lipid peroxidation assay by thiobarbituric acid-reactive substances (TBARS) method using rat brain homogenate. The results indicate that D. gangeticum extract has potent antioxidant activity. PMID- 14519949 TI - Sauchinone, a lignan from Saururus chinensis, inhibits staurosporine-induced apoptosis in C6 rat glioma cells. AB - Neuronal apoptosis may contribute to pathologic neuronal loss in certain disease states such as neurodegenerative diseases. Staurosporine (ST), a nonselective protein kinase inhibitor, has been shown to induce apoptosis in a variety of cells including nerve cell lines. In this study, we investigated the neuroprotective effect of sauchinone, which is a unique lignan from Saururus chinensis, on ST-induced apoptosis in C6 rat glioma cells. Sauchinone attenuated ST-induced apoptosis of C6 glioma cells as evidenced by DNA fragmentation. We also provide evidence that the inhibitory effect of sauchinone on ST-induced apoptosis involves a dose-dependent upregulation of an antiapoptotic protein, Bcl 2. Mounting evidence shows that the activation of caspases, especially caspase-3, triggers the apoptotic process. The activity of caspase-3 of ST-pretreated cells was significantly decreased upon sauchinone treatment in a dose-dependent manner. Taken together, the data demonstrate that sauchinone protects C6 glioma cells from ST-induced apoptosis in a caspase-3 dependent manner. Our findings may be critical for developing a strategy to protect nerve cells from apoptosis, suggesting the potential development of sauchinone as a neuroprotective agent. PMID- 14519950 TI - Antiproliferative activity of cardenolides isolated from Streptocaulon juventas. AB - Sixteen cardenolides, two hemiterpenoids, two phenylpropanoids and a phenylethanoid isolated from the roots of Streptocaulon juventas (LOUR.) MERR. were examined for their antiproliferative activity toward three human-derived (HT 1080 fibrosarcoma, lung A549 adenocarcinoma, cervix HeLa adenocarcinoma) and three murine-derived (colon 26-L5 carcinoma, Lewis lung carcinoma, B16-BL6 melanoma) cell lines. The cardenolides selectively and strongly inhibited proliferation of the HT-1080 (IC(50) values, 0.054-1.6 microM) and A549 (IC(50), 0.016-0.65 microM) cell lines. The characteristic morphological changes and ladder-like DNA fragmentation in those cells treated with the cardenolides indicated the antiproliferative activity was due to the induction of apoptosis. PMID- 14519951 TI - Antinociceptive and antiinflammatory effects of Niga-ichigoside F1 and 23 hydroxytormentic acid obtained from Rubus coreanus. AB - As an attempt to search for bioactive natural constituents exerting antinociceptive and antiinflammatory activities, we examined the potency of the extract of Rubus coreanus fruits by the activity-guided fractionation. The EtOAc- and BuOH fraction and those alkaline hydrolysates showed significant antinociceptive effects as assessed by writhing-, hot plate- and tail flicks tests in mice and rats as well as antiinflammatory effect in rats with carrageenan-induced edema. BuOH extract was subjected to column chromatography to obtain a large amount of niga-ichigoside F(1) (1,23-hydroxytormentic acid 28-O glc), which was again hydrolyzed in NaOH solution to yield an aglycone 23 hydroxytormentic acid (1a). The aglycone, 23-hydroxytormentic acid, was much more potent in both antinociceptive and antiinflammatory tests than the glycoside, niga-ichigoside F(1). The antiinflammatory effects of these compounds were further supported by the reduction of carrageenan-induced lipid peroxidation and hydroxyl radical in serum. These results suggested that 23-hydroxytormentic acid might be an active moiety of niga-ichigoside F(1) present in R. coreanus. PMID- 14519952 TI - Suitability of the cynomolgus monkey as an animal model for drug absorption studies of oral dosage forms from the viewpoint of gastrointestinal physiology. AB - To determine the usefulness of the monkey as an animal model, which can predict in vivo performance of humans, the major gastrointestinal physiological parameters of this animal were evaluated. The pH of gastric juice collected by a fiberscope from the stomach in fasted cynomolgus monkeys showed a high acidity level, which ranged from 1.2 to 4.3. The gastric emptying time of oral dosage forms (solution, granules and tablets) showed that the larger size dosage forms seemed to be emptied more slowly, and three dosage forms were prolonged by feeding. The gastrointestinal agitation intensity of monkeys was estimated using controlled-release tablets of acetoaminophen, which showed a slow erosion rate. The in vivo release amount-time profiles of the tablet in fasted monkeys corresponded to their in vitro profiles with paddle agitation conditions of between 10 rpm and 50 rpm of the paddle method; this result was smaller than in dogs (100 rpm) but equivalent to that in humans (10 rpm). Further, the small intestinal transit time (SITT), estimated using a double marker method, ranged from 2.2 to 4.2 h in the fasting state and from 2.2 to 3.2 h in the fed state; the SITT was not significantly delayed by feeding. Comparison with the published data about dogs and humans showed these gastrointestinal physiological parameters of monkeys to be more similar to those of humans. Consequently, it is assumed that the monkey is useful as an animal model for bioavailability studies of oral dosage forms. PMID- 14519953 TI - 4'-hydroxylation of flurbiprofen by rat liver microsomes in fasting and feeding conditions. AB - We examined the 4'-hydroxylation of flurbiprofen in rat hepatocytes and liver microsomes in order to know whether the metabolism of flurbiprofen is changed on its administration to experimental animals after overnight fasting, because starvation and fasting change both the composition of cytochrome P450s (CYPs) and metabolic activity. CYPs involved in the hydroxylation were determined by various CYP inhibitors and inhibitory antibodies against rat CYP2C11 and CYP2E1 using the microsomes in fasting and feeding. The results provided a possibiliy that the 4' hydroxylation might be regulated by CYP2C11, but not by CYP2E1, at fasting rather than feeding. PMID- 14519954 TI - Metabolite profiles of flutamide in serum from patients with flutamide-induced hepatic dysfunction. AB - Hepatic dysfunction due to flutamide administration has been reported and this side effect often limits the use of the agent. The prediction of flutamide induced hepatotoxicity is attributed to the proper use of the antiandrogen. In this study, we investigated whether hepatic dysfunction could be assessed by the metabolite profile in serum from patients receiving this drug. Serum samples were obtained from 15 patients with prostate cancer, 12 patients with no sign of hepatotoxicity and 3 patients with slight hepatic dysfunction during long-term flutamide treatment. We analyzed the metabolite profiles by LC/MS in selected ion monitoring mode and detected a new metabolite (M3) that was an oxidation product of flutamide. However, there were no consistent differences in the serum flutamide metabolites between patients with normal function and those suffering hepatic dysfunction. The metabolite profiles in the beta-glucuronidase-treated serum showed a similar pattern between normal functioning and dysfunctional groups. Thus, the profile of flutamide metabolites determined in serum may not contribute to the risk prediction of flutamide-related hepatotoxicity. PMID- 14519955 TI - Intradermal delivery of antisense oligonucleotides by the pulse depolarization iontophoretic system. AB - The intradermal delivery of an antisense oligonucleotide was examined by iontophoresis. In this experiment, the antisense sequence of [(32)P]-labeled phosphodiester oligonucleotide ([(32)P]D-oligo, 18-mer) hybridizing to mouse interleukin 10 (IL-10) mRNA was used as a model D-oligo. In in vitro iontophoretic experiments, isolated hairless mouse skin was used with a horizontal diffusion cell. The enhancing effect of pulse depolarization (PDP) iontophoresis on the [(32)P]D-oligo permeation through the skin was better, and the skin irritation was less, than those of constant direct current (CDC) iontophoresis. The apparent fluxes of [(32)P]D-oligo were enhanced with the increasing current densities and [(32)P]D-oligo concentrations in the donor solution, whereas the enhanced flux decreased with the increasing NaCl concentrations in the donor solution. An optimum electric current was observed for the intradermal delivery of [(32)P]D-oligo, and intact [(32)P]D-oligo was detected within the skin after iontophoresis for 6 h. These results suggest that PDP iontophoresis may be useful for the intradermal delivery of antisense oligonucleotides. PMID- 14519956 TI - Micropropagation of Polygonum multiflorum THUNB and quantitative analysis of the anthraquinones emodin and physcion formed in in vitro propagated shoots and plants. AB - An efficient and rapid protocol for in vitro induction and complete plant regeneration of Polygonum multiflorum THUNB has been developed. Nodal explants were grown in vitro on Murashige and Skoog's (MS) basal medium containing different concentrations of alpha-naphthaleneacetic acid (NAA) and benzyladenine (BA). The nodal explants (97%) produced multiple shoots (4.7 shoots per explant) on MS basal medium supplemented with 0.2 mg/l NAA and 2.0 mg/l BA after 6 weeks of culture. Eighty-eight percent to 100% of the shoots (1.0 cm in length) elongated (about 3.02-4.28 cm) and rooted on MS basal medium supplemented with NAA or indole-3-butyric acid (IBA). All the rooted shoots were transferred to pots containing autoclaved soil, vermiculite, and peat moss (1 : 1 : 1). The plantlets were successfully acclimatized under greenhouse conditions with high humidity before transferring to the field. The anthraquinone contents were determined using HPLC. Analysis revealed that the contents of the major medicinal compounds-emodin and physcion in the 6 weeks old in vitro grown shoots and three month old in vitro propagated plants grown in greenhouse were higher than those of the marketed crude drug (processed underground or stem parts of P. multiflorum). PMID- 14519957 TI - Therapeutic effects of Hominis placenta injection into an acupuncture point on the inflammatory responses in subchondral bone region of adjuvant-induced polyarthritic rat. AB - Rheumatoid arthritis (RA) is characterized by chronic inflammation of the synovial membrane in the joint, which leads to the progressive destruction of articular cartilage, ligament and bone. Several cytokines such as tumor necrosis factor alpha (TNF-alpha), interleukin 1beta (IL-1beta), and interleukin 6 (IL-6) have been implicated in the pathological mechanisms of synovial tissue proliferation, joint destruction and programmed cell death in rheumatoid joint. In the Korean traditional medicine, Hominis placenta (HP) as an herbal component of herb-acupuncture has been widely used to treat chronic inflammatory diseases such as RA. To study the therapeutic effects of HP injection into the ST36 acupoint (HP herb-acupuncture) on the inflammatory responses of a subchondral region of rheumatoid joint, the polyarthritis-induced Sprague-Dawley (SD) rat was developed as a rheumatoid arthritis model by the intradermal injection of dried cells of Mycobacterium tuberculosis emulsified in squalene to the base of tail. After the onset stage (11 d after adjuvant injection) of polyarthritis, a fixed volume of HP extract was daily injected to Zusanli (ST36) acupoint on the rat's leg for 2 weeks. The body weight, paw volume of the knee joint and articular index were exploited as an assessment method addressing arthritic symptoms, and the expression profiles of TNF-alpha, IL-1beta and IL-6 at the subchondral bone of the joint were analyzed using an immunohistochemistry. After the treatment of arthritic rats with HP, the body weights and paw volumes of arthritic rats were almost restored to the levels of normal rats whereas the evaluation by the articular index was not remarkable. The TNF-alpha, IL-1beta and IL-6 positive cells in the immunohistological sections of subchondral bone region of the joint significantly decreased in HP-treated (ST36 acupoint) arthritic group as compared with those in non-treated or HP-treated (non-acupoint) ones, which was coincident with the behavioral studies. In conclusion, the HP herb-acupuncture was found to be effective to alleviate the arthritic symptoms in adjuvant-induced arthritis rats as regards the body weight, joint appearance and the expression profiles of inflammatory cytokines. PMID- 14519958 TI - Flavone markedly affects phenotypic expression of beta-lactam resistance in methicillin-resistant Staphylococcus aureus strains isolated clinically. AB - Flavone and its derivatives had very weak antibacterial effects on methicillin resistant Staphylococcus aureus (MRSA) and methicillin-sensitive S. aureus, but dramatically intensified MRSA's susceptibility to beta-lactams. We named these compounds "ILSMR (intensifier of beta-lactam-susceptibility in MRSA)." We also found discrepancies among MRSA strains in their responses to flavone; some strains showed phenotypic susceptibility to methicillin while others showed phenotypic resistance to it. To understand the mechanism underlying this discrepancy, we characterized 20 MRSA strains in detail, analyzed their conventional and molecular typings, and examined each strain's resistance to beta lactams, with COL serving as a reference. Neither SCCmec typing nor coagulase typing explained the diverse effects of flavone on the beta-lactam MICs of these strains. Likewise, changes in pulsed-field gel electrophoresis (PFGE) type were not associated with the profiles of ILSMR effects. However, the present observations suggest that the ILSMR effects on MRSA is strain-specific, and that this effect depends on an as-yet unknown mechanism that is essential for the expression of the phenotype conferring beta-lactam resistance to MRSA strains, independently of an interaction with the mecA-encoded penicillin-binding protein 2a or with the beta-lactamase. PMID- 14519959 TI - Probucol decreases mevalonate pyrophosphate decarboxylase in the rat liver. AB - It is known that cholesterol biosynthesis in the liver is inhibited by probucol. This inhibition by probucol is caused at least in part by a decrease in 3-hydroxy 3-methylglutaryl (HMG)-CoA reductase activity. In this study, we examined serum cholesterol and the change in the activity or protein level of mevalonate pyrophosphate decarboxylase (MPD), which is involved in cholesterol biosynthesis, in the livers of rats fed probucol. The results indicated that serum cholesterol, MPD activity and MPD protein were decreased by 70, 50 and 60% by probucol, respectively, as compared with those in rats fed normal chow. These data show for the first time that probucol decreases the level of an enzyme involved in cholesterol biosynthesis other than HMG-CoA reductase. PMID- 14519961 TI - Effects of insulin and adrenalectomy on elevation of serum leptin levels induced by 5-hydroxytryptophan in mice. AB - We previously reported that a serotonin precursor 5-hydroxytryptophan (5-HTP) increases serum leptin levels in mice. In this study, we studied the effects of insulin and adrenalectomy on hyperleptinemia induced by 5-HTP. Co-administration of insulin significantly increased hyperleptinemia elicited by 5-HTP. 5-HTP itself increased serum insulin levels. Adrenalectomy, which depletes corticosterone, did not abolish hyperleptinemic effects of 5-HTP. These results suggest that insulin may participate in hyperleptinemic effects of 5-HTP and that the involvement of corticosterone in effects of 5-HTP may be probably small. PMID- 14519960 TI - Biological activity of tropolone. AB - Tropolone (1). showed strong insecticidal activity on Tyrophagus putrescentiae and Dermatophagoides farinae. The insecticidal effect of 1 on both insects was stronger than that of hinokitiol (2, 4-isopropyltropolone: major component of Thujopsis dolabrata SIEB. et ZUCC. hondai MAKINO). The insecticidal activity of both compounds was higher than that of N,N-diethyl-m-toluamide (DEET), used as a positive control. Compound 1 had potent insecticidal activity against Coptotermes formosanus, although its activity was much lower than that of commercial chloropyrifos. Like 2, 1 showed the inhibitory activity toward metalloproteases such as carboxypeptidase A, collagenase and thermolysin and their inhibitory activities were much higher than that of 1,10-phenanthroline, used as a positive control. The inhibitory activity of 1 on carboxypeptidase A was especially high, its 50% inhibitory concentrations (IC(50)) being 2.73 x 10(-6) M. This inhibitory activity was as high as that of 2 (IC(50): 2.76 x 10(-6) M). Compound 1 inhibited the growth of seven kinds of plant-pathogenic fungi and their minimum inhibitory concentration (MIC) values were in the range of 6.0-50.0 microg/ml. In particular, 1 showed strong antifungal activity on Pythium aphanidermatum IFO 32440 (MIC: 6.0 microg/ml). PMID- 14519962 TI - Levels of some acute-phase proteins in the serum of patients with cancer during radiotherapy. AB - The acute phase response involves changes in serum concentrations of a number of liver-synthesized proteins. Among these are C-reactive protein (CRP), ferritin (FER), transferrin (Trf) and ceruloplasmin (Cp). Determination of serum CRP, FER, Trf, and Cp was performed in 52 patients with inoperable head and neck cancer (n=11), inoperable esophageal cancer (n=10), rectal cancer (n=9; operation was performed=5, inoperable=4), and lung cancer (n=22), all of whom were treated with radical radiotherapy (RT). Post-radiotherapy CRP levels were significantly higher compared to the preradiotherapy levels (p<0.001). We found decreased serum Trf levels during the irradiation period, while acute-phase proteins such as CRP, FER, and Cp levels increased during the RT period. Further studies on the roles of other acute phase reactants and the above mentioned parameters in a large patients-with cancer group during radiotherapy are required to understand the role of markers, which are altered during radiotherapy. PMID- 14519963 TI - Anti-HIV active petrosins from the marine sponge Petrosia similis. AB - A marine sponge Petrosia similis afforded two compounds which belongs to bis quinolizidine alkaloids namely, petrosin (1) and petrosin-A (2), respectively. Petrosin (1) and petrosin-A (2) showed anti-HIV inhibition with IC(50) values of 41.3 and 52.9 microm respectively. MAGI cell assays indicated that the compounds inhibited early steps of HIV replication. In extracellular HIV-1 Reverse Transcriptase inhibition assay petrosin and petrosin-A inhibited HIV-1 RT at 10.6 and 14.8 microm. This is the first report of petroisns with anti-HIV activity. PMID- 14519965 TI - Effects on blood pressure decrease in response to PAF of Impatiens textori MIQ. AB - A 35% EtOH extract of flowers of Impatiens textori MIQ. showed an inhibitory effect on blood pressure decrease in response to platelet activating factor (PAF) measured with a blood pressure monitoring system. Bioassay-guided fractionation of the 35% EtOH extract (IT) led to isolation of the flavones apigenin (1) and luteolin (3), which significantly inhibited blood pressure decrease in response to PAF. Their compounds and apigenin 7-glucoside (2), chrysoeriol (4), quercetin (5), quercetin 3-glucoside (6), kaempferol (7), kaempferol 3-glucoside (8) and kaempferol 3-rhamnosyldiglucoside (9) were also isolated from the flowers of I. textori for the first time. This study revealed that the flowers of I. textori might be a possible anti-allergy agent. PMID- 14519964 TI - Ethanol fraction of Aralia elata Seemann enhances antioxidant activity and lowers serum lipids in rats when administered with benzo(a)pyrene. AB - Aralia elata Seemann is an edible mountain vegetable in Korea containing saponin, alkaloid, palmitic acid, linoleic acid, methyl eicosanoate and hexacosol, and is known to manifest an effect on cardiac infarction, gastric ulcer, colitis, and enervation. This study has examined the effects of Aralia elata Seemann ethanol extract on antioxidant enzyme systems and lipid metabolism in rats along with benzo(a)pyrene (B(a)P) administration. Rats were divided into four groups: control (C), an extract fed group (CE), a B(a)P fed group (CB), and a B(a)P and extract fed group (CBE). The ethanol extracts of Aralia elata Seemann (50 mg/kg body weight) were fed to the rats for 4 weeks by stomach tubing. Extract administration increased the antioxidant activities of glutathione sulfur transferase (GST). Total superoxide dismutase (SOD) and Cu,Zn-SOD activities were stimulated. Catalase activities were increased by 50% with extract feeding. Cu,Zn SOD was greatly enhanced from 0.10 unit to 0.18 unit and catalase activity also was increased. Serum alpha-tocopherol was markedly increased by the extracts. The ethanol fraction of Aralia elata Seemann decreased total serum cholesterol. However, serum HDL-cholesterol was increased by 35% (p<0.05). The results indicate that Aralia elata Seemann exerts antioxidant and strong hypocholesterolemic and hypolipidemic effects in vivo with the administration of B(a)P. PMID- 14519966 TI - The effects of calcium chloride and sodium chloride on the electroporation mediated skin permeation of fluorescein isothiocyanate (FITC)-dextrans in vitro. AB - We previously reported the substantial synergic effects of electroporation and electrolytes, particularly those containing CaCl(2) on the skin permeation of the model low-molecular weight compound, calcein. We then investigated the effects of electroporation (300 V, 10 msx10 times) and 150 mM NaCl or CaCl(2) on skin permeation of higher molecular weight compounds, fluorescein isothiocyanate (FITC)-dextrans (FD-4, FD-10 and FD-40; average molecular weight, 4.4, 9.6 and 35.6 kDa, respectively) using excised hairless rat skin. The observed steady state flux of FD-4 was 1.3 pmol/cm(2)/h after electroporation without NaCl or CaCl(2). The flux did not differ greatly from that without electroporation. In contrast, a much higher steady state flux was observed after electroporation with NaCl or CaCl(2) (2.5 and 8.2 pmol/cm(2)/h, respectively). For FD-10 and FD-40, no flux was detected with electroporation in water (without electrolytes) or without electroporation. On the other hand, high skin permeation was observed after electroporation in NaCl or CaCl(2) solution (FD-10: 7.5 and 18.2 pmol/cm(2)/h, FD 40: 4.5 and 9.3 pmol/cm(2)/h in NaCl and CaCl(2), respectively). The effects of CaCl(2) on FD permeation were greater than those of NaCl. The present finding suggests that electroporation application in the presence of electrolytes, particularly CaCl(2), was very effective in increasing transdermal delivery of water-soluble macromolecules. PMID- 14519967 TI - Suppression of ethanol and lipopolysaccharide-induced liver injury by extracts of Hydrangeae Dulcis Folium in rats. AB - In female SD rats that were injected with 4 g/kg BW ethanol p.o. followed by a 5 mg/kg BW lipopolysaccharide (LPS) i.v. injection, serum glutamic pyruvic transaminases (GPT) activity increased to about eight times that of normal rats. In this model, rats that had been fed a diet containing 1% Hydrangeae Dulcis Folium (HDF) extracts for fifteen days showed significantly lower serum GPT activity (380.0+/-58.2 IU/l) than the control group (3527.0+/-774.1 IU/l). HDF's efficacy was far superior to milk thistle in this model (2950.0+/-915.9 IU/l). When mouse macrophages were treated with HDF extracts at 50 microg/ml, TNF-alpha production induced by LPS was suppressed to about 10% of the control. Rat serum TNF-alpha levels induced by LPS was decreased to 58.7% of the control by administering 1000 mg/kg BW HDF extract p.o. These results indicate that HDF prevents alcohol-induced liver injury through the inhibition of TNF-alpha production. PMID- 14519968 TI - Microencapsulation of linoleic acid with low- and high-molecular-weight components of soluble soybean polysaccharide and its oxidation process. AB - Soluble soybean polysaccharide (SSPS) was fractionated into its low- (LMW) and high-molecular-weight (HMW) components to test their antioxidative and emulsifying properties. Linoleic acid was emulsified with an aqueous solution of SSPS, HMW, a mixture of LMW or HMW with maltodextrin, or maltodextrin alone. The emulsions prepared with SSPS, HWM and the mixture of HMW with maltodextrin were stable. These emulsions were spay-dried to produce microcapsules. The encapsulated linoleic acid was oxidized at 37 degrees C and at various levels of relative humidity. Linoleic acid encapsulated with the mixture of LMW with maltodextrin or HMW was stable to oxidation, and this stability increased as the weight fraction of LMW in the mixture was increased. The LMW components also had high DPPH-radical scavenging activity. These results indicate that LMW played an important role in suppressing or retarding the oxidation of linoleic acid encapsulated with SSPS. The oxidative stability of linoleic acid encapsulated with a mixture of the LMW and HMW components was high at low and high relative humidity, but not at intermediate levels of relative humidity. PMID- 14519969 TI - Oxidative reaction of oxindole-3-acetic acids. AB - The oxindole-3-acetic acids, oxidative metabolites of indole-3-acetic acid, were isolated from a byproduct of a corn starch manufacturing plant, and were further converted to the 3-hydroxyl derivatives in the presence of metal ion. The mechanical study was followed by a chemical analysis including other byproducts, and suggested the presence of an intermediate that had a radical at the C-3 position of oxindole-3-acetic acids. PMID- 14519970 TI - Characterization and kinetics of 45 kDa chitosanase from Bacillus sp. P16. AB - An extracellular 45 kDa endochitosanase was purified and characterized from the culture supernatant of Bacillus sp. P16. The purified enzyme showed an optimum pH of 5.5 and optimum temperature of 60 degrees C, and was stable between pH 4.5 10.0 and under 50 degrees C. The Km and Vmax were measured with a chitosan of a D.A. of 20.2% as 0.52 mg/ml and 7.71 x 10(-6) mol/sec/mg protein, respectively. The enzyme did not degrade chitin, cellulose, or starch. The chitosanase digested partially N-acetylated chitosans, with maximum activity for 15-30% and lesser activity for 0-15% acetylated chitosan. The chitosanase rapidly reduced the viscosity of chitosan solutions at a very early stage of reaction, suggesting the endotype of cleavage in polymeric chitosan chains. The chitosanase hydrolyzed (GlcN)7 in an endo-splitting manner producing a mixture of (GlcN)(2-5). Time course studies showed a decrease in the rate of substrate degradation from (GlcN)7 to (GlcN)6 to (GlcN)5, as indicated by the apparent first order rate constants, k1 values, of 4.98 x 10(-4), 2.3 x 10(-4), and 9.3 x 10(-6) sec(-1), respectively. The enzyme hardly catalyzed degradation of chitooligomers smaller than the pentamer. PMID- 14519971 TI - Apoptosis induction in HL-60 cells and inhibition of topoisomerase II by triterpene celastrol. AB - Celastrol, which is a triterpene purified from Celastraceae plants, has anticancer and anti-inflammatory activities. In this study we investigated to clarify whether celastrol can induce apoptosis in a human leukemia HL-60 model system. Celastrol was found to induce apoptosis, and the rank order of the potency of celastrol and its derivatives to induce internucleosomal DNA fragmentation was found to be celastrol>cela-H>>the other derivatives=vehicle control. Many anticancer agents are known to possess the ability to inhibit topoisomerase II, so the inhibitory activities of celastrol and its derivatives on topoisomerase II were also explored. The rank order of the inhibitory activity was found to be celastrol>etoposide>cela-H, indicating that the apoptosis inducing activities of cela derivatives correspond to their inhibitory activities on topoisomerase II. These data suggested that celastrol may cause its effects such as anticancer activity by the mechanism of apoptosis along with topoisomerase II inhibition. PMID- 14519972 TI - Purification and characterization of two isoforms of glucose 6-phosphate dehydrogenase (G6PDH) from Chlorella vulgaris C-27. AB - Two kinds of isoforms of glucose 6-phosphate dehydrogenase (G6PDH) were purified from cells of a freezing-tolerant strain, Chlorella vulgaris C-27, by sequential steps of chromatography on five kinds of columns, including a HiTrap Blue column which showed excellent separation of the isoforms from each other. The two isoforms (G6PDH1 and G6PDH2) were purified up to 109-fold and 197-fold with specific activity of 14.4 and 26.0 U/mg-protein, respectively. G6PDH1 showed an apparent Mr of 200,000 with a subunit Mr of about 58,000, whereas G6PDH2 showed an apparent Mr of 450,000 with a subunit Mr of about 52,000. The kinetic parameters were measured and several enzymatic features of the isoforms, such as effects of metal ions on the enzyme activity, were clarified, which showed that the two isoforms were different from each other in many respects. Among the effective ions, Cd2+ showed marked stimulating effects on both isoforms. G6PDH1 and G6PDH2 seem to be a cytosolic and a chloroplastic type, respectively, as judged by their sensitivity to DTT, and also from the results of sequence similarity searches using their N-terminal and internal amino acid sequences. PMID- 14519973 TI - Improved functional properties of the ovoinhibitor by conjugating with galactomannan. AB - The chicken egg white ovoinhibitor, a multi-type proteinase inhibitor, was conjugated with galactomannan through the Maillard reaction in a controlled dry heating state at 60 degrees C and 65% relative humidity. The formation of an ovoinhibitor-galactomannan conjugate during dry heating was confirmed by SDS PAGE. The resulting ovoinhibitor-galactomannan conjugate showed almost the same inhibitory activity toward trypsin, chymotrypsin and elastase as that of the untreated ovoinhibitor, while the conjugate showed stronger heat stability and better emulsifying properties than the untreated ovoinhibitor. These results suggest that the ovoinhibitor-galactomannan conjugate can be used as a protease inhibitor having heat stability and outstanding emulsifying properties for industrial application. PMID- 14519974 TI - Metabolism of deuterium-labeled jasmonic acid and OPC 8:0 in the potato plant (Solanum tuberosum L.). AB - The metabolism of deuterium-labeled (+/-)-jasmonicacid and 3-oxo-2-[(Z)pent-2' enyl]-cyclopentan-1- octanoic acid in potato (Solanum tuberosum L.) was examined by using cultures of potato single-node stems. Deuterium-labeled (+/-)-jasmonic acid and 3-oxo-2-[(Z) pent-2'-enyl]cyclopentan-1-octanoic acid, which had been prepared from commercially available methyl (+/-)-jasmonate, were fed to the cultures, and the metabolites were extracted from the plants and analyzed by a liquid chromatography-selected ion monitoring system. The metabolism of deuterium labeled (+/-)-jasmonic acid and 3-oxo-2-[(Z)-2'-pentenyl]cyclopentan-1-octanoic acid to 5' and 4'-O-glucopyranosyloxyjasmoic acids was strongly suggested. PMID- 14519975 TI - Malformation of immature starfish oocytes by theonellapeptolide Ie, a Tridecapeptide lactone from a marine sponge Petrosia species, through disturbance of cortical F-actin distribution. AB - Theonellapeptolide Ie (Tp), an oligopeptide lactone isolated from a marine sponge, Petrosia sp., was shown to induce an unprecedented morphological change in the immature oocytes of the starfish Asterina pectinifera. The cortical F actin was disturbed and assembled to form dots and rings, as evidenced by staining with rhodamine-conjugated phalloidin. The oocyte eventually became malformed. When Tp was added to an immature oocyte which had been pretreated with cytochalasin B or D, inhibitors of actin polymerization, no malformation was observed. When Tp was added to an oocyte which had been induced to mature by 1 methyladenine (1-MeAde), a maturation-inducing substance in starfishes, no morphological changes were observed in the maturing oocytes which reached the first meiotic prometaphase 40 min after the start of 1-MeAde treatment. This is the first description of a chemical that induces aberrant redistribution of F actin-based cytoskeleton in an animal oocyte which is arrested at the first meiotic prophase. PMID- 14519976 TI - Suppressive effects of genistein on oxidative stress and NFkappaB activation in RAW 264.7 macrophages. AB - This study was designed to investigate whether genistein may ameliorate oxidative stress and nuclear factor kappaB (NFkappaB) activation in the lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophage cell line. Treatment of RAW 264.7 cells with genistein significantly reduced lipopolysaccharide (LPS)-stimulated nitric oxide (NO) production in a dose-dependent manner with an IC50 of 69.4 microM. Genistein at 50 microM and 100 microM concentrations reduced thiobarbituric acid-reactive substances (TBARS) accumulation, increasing the GSH level and antioxidant enzyme activities, such as superoxide dismutase (SOD) and catalase. The specific DNA-binding activities of nuclear factor kappaB (NFkappaB) on nuclear extracts from 50 microM and 100 microM genistein treatments were significantly suppressed. These results suggest that genistein has mild antioxidant activity to suppress intracellular oxidative stress and NFkappaB activation. PMID- 14519977 TI - Production of D-arabitol by Metschnikowia reukaufii AJ14787. AB - A potent producer of D-arabitol was isolated by screening of natural sources and identified as Metschnikowia reukaufii AJ14787. Resting cells of this strain can efficiently produce D-arabitol from D-glucose with a weight yield of more than 60%, and can also produce D-arabitol from several other types of sugars such as polyols, ketoses, and aldoses. To improve productivity, various culture conditions such as temperature and the concentrations of D-glucose and nitrogen sources were examined. Under optimal conditions, 206 g/l of D-arabitol was produced from D-glucose with a weight yield of 52% in 100 hours. PMID- 14519978 TI - Reduction of noise-stress-induced physiological damage by radices of Astragali and Rhodiolae: glycogen, lactic acid and cholesterol contents in liver of the rat. AB - Noise is one of the factors that induces critical stress in animals. The contents of glycogen, lactic acid and cholesterol in the liver of noise-stressed rats were analyzed in order to investigate the alleviation of noise-stress-induced physiological damages by traditional medicine using Astragali and Rhodiolae radices. More than 95 dB noise ranging from 2 to 4 kHz reduced the contents of these compounds in the liver of rats not injected with the extract of Astragali or Rhodiolae, but did not change the contents in the liver of rats injected with the Astragali or Rhodiolae extract. These results show that noise induced stress in the rats via a decrease in contents of these compounds in the liver and that Astragali or Rhodiolae maintained the contents of these compounds in the liver of the noise-stressed rats. The results indicate that Astragali or Rhodiolae improved the ability for rats to resist noise stress. PMID- 14519979 TI - Effects of highly purified structured lipids containing medium-chain fatty acids and linoleic acid on lipid profiles in rats. AB - The purpose of this study is to examine the effects of highly purified structured lipids on serum and liver lipid profiles in rats. We also investigated in vitro hydrolysis of lipid emulsions by porcine pancreas. Hydrolysis rates of medium chain (M)-linoleic (L)-medium chain (M) types were 2 to 3 times higher than those of L-M-L types. The diet containing structured lipids or corn oil was administered to rats for 4 weeks. There were no significant differences in growth and food efficiency. Serum cholesterol levels were significantly lower (P<0.05) in the 2-octanoyl-1,3-dilinoleoyl-glycerol, 2-linoleoyl-1,3-didecanoyl-glycerol, and 2-decanoyl-1,3-dilinoleoyl-glycerol groups than in the corn-oil group. Serum triglyceride levels were significantly lower (P<0.05) in rats fed L-M-L types than those in the other groups. Serum non-esterified fatty acid (NEFA) and beta hydroxybutylate levels were significantly higher (P<0.01) in rats fed M-L-M types than those of the other groups. These results indicate that the feeding of highly purified L-M-L types could effectively improve serum and liver lipid profiles and that M-L-M types may be a preferable substrate for the pancreas and contribute to energy supply in rats. PMID- 14519980 TI - Cytotoxicity and DNA topoisomerase inhibitory activity of benz[f]indole-4,9-dione analogs. AB - A series of benz[f]indole-4,9-diones, based on the antitumor activity of 1,4 naphthoquinone, were synthesized and evaluated for their cytotoxic activity in cultured human cancer cell lines A549 (lung cancer), Col2 (colon cancer), and SNU 638 (stomach cancer), and also for the inhibition of human DNA topoisomerases I and II activity in vitro. Several compounds including 2-amino-3-ethoxycarbonyl-N methyl-benz[f]indole-4,9-dione showed a potential cytotoxic activity judged by IC50<20.0 microg/ml in the panel of cancer cell lines. Especially, 2-hydroxy-3 ethoxycarbonyl-N-(3,4-dimethylphenyl)-benz[f]indole-4,9-dione had potential selective cytotoxicity against lung cancer cells (IC50=0.4 microg/ml)) compared to colon (IC50>20.0 microg/ml) and stomach (IC50>20.0 microg/ml) cancer cells. To further investigate the cytotoxic mechanism, the effects of test compounds on DNA topoisomerase I and II activities were used. In a topoisomerase I-mediated relaxation assay using human placenta DNA topoisomerase I and supercoiled pHOTI plasmid DNA, 2-amino-3-ethoxycarbonyl-N-(4-fluorophenyl)-benz[f]indole-4,9-dione had the most potent inhibitory activity among the compounds tested. However, most of the compounds showed only weak inhibition of the DNA topoisomerase II-mediated KDNA (Kinetoplast DNA) decatenation assay, except for 2-amino-3-ethoxycarbonyl-N (4-methylphenyl)-benz[f]indole-4,9-dione and 2-amino-3-ethoxycarbonyl-N-(2 bromoehtyl)-benz[f]indole-4,9-dione with a moderate inhibitory activity. These results suggest that several active compounds had relatively selective inhibitory activity against toposiomearse I compared to toposiomerase II. No obvious correlation was observed between the cytotoxicity of the individual compound and the inhibitory activity of DNA relaxation and decatenation by topoisomerase I and II, respectively, in vitro. PMID- 14519982 TI - New method for synthesizing the intermediates to 5-HETE from yeast-mediated reduction products by employing Baeyer-Villiger oxidation with complete retention of enantiomeric excess. AB - (R) and (S)-Aldehydes 2, which are intermediates for the synthesis of (5R) and (5S)-HETE, were respectively synthesized from the yeast-mediated reductive products, hydroxy ester 3 and cis-lactone 4, through Baeyer-Villiger oxidation with complete retention of enantiomeric excess. PMID- 14519981 TI - Properties of a novel extracellular cell-free ice nuclei from ice-nucleating Pseudomonas antarctica IN-74. AB - Some ice-nucleating bacterial strains, including Pantoea ananatis (Erwinia uredovora), Pseudomonas fluorescens, and Pseudomonas syringae isolates, were examined for the ability to shed ice nuclei into the growth medium. A novel ice nucleating bacterium, Pseudomonas antarctica IN-74, was isolated from Ross Island, Antarctica. Cell-free ice nuclei from P. antarctica IN-74 were different from the conventional cell-free ice nuclei and showed a unique characterization. Cell-free ice nuclei were purified by centrifugation, filtration (0.45 microm), ultrafiltration, and gel filtration. In an ice-nucleating medium in 1 liter of cell culture, maximum growth was obtained with the production of 1.9 mg of cell free ice nuclei. Ice nucleation activity in these cell-free ice nuclei preparations was extremely sensitive to pH. It was demonstrated that the components of cell-free ice nuclei were protein (33%), saccharide (12%), and lipid (55%), indicating that cell-free ice nuclei were lipoglycoproteins. Also, carbohydrate and lipid stains showed that cell-free ice nuclei contained both carbohydrate and lipid moieties. PMID- 14519983 TI - The dual functions of biphenyl-degrading ability of Pseudomonas sp. KKS102: energy acquisition and substrate detoxification. AB - The bph operon of Pseudomonas sp. KKS102 is constituted of 11 bph genes which encode enzymes for biphenyl assimilation. Growth of a mutant in which a large part of the bph operon was deleted was inhibited by biphenyl in a concentration dependent manner. We constructed a series of bph operon deletion mutants and tested for their biphenyl sensitivity. Growth inhibition by biphenyl was more prominent with the mutants defective in bphA1, bphB, bphC, and bphD, which were clustered in the bph operon and working in the early stage of the biphenyl degradation. The mutant defective in bphE, which was working at the late stage and forming a different cluster from the early stage genes, was not much inhibited by biphenyl. These indicate that biphenyl is detoxified by enzymes which function in the early stage of biphenyl assimilation and thus detoxification of substrates as well as energy acquisition could have played an important role in the evolution of the KKS102 bph operon. PMID- 14519984 TI - Occurrence of a specific protein in basidiomycete-lytic enzyme preparation produced by Bacillus circulans KA-304 inductively with a cell-wall preparation of Schizophyllum commune. AB - KA-prep, a culture filtrate of Bacillus circulans KA-304 grown on a cell-wall preparation (CWP) of Schizophyllum commune, has been reported to have an activity to form protoplasts from S. commune mycelia. The SDS-polyacrylamide gel electrophoreses described here demonstrated that a specific proteinous component (molecular weight: 150,000) occurred in KA-prep. The protein (P150T) was also formed in culture filtrates with CWP of several basidiomycetes, which could release the protoplasts, suggesting that the component was an indispensable factor for protoplast formation. P150T, isolated from an ammonium sulfate fraction of KA-prep (0-30% saturation), did not have any protoplast-forming activity. Results were obtained indicating that P150T participates in protoplast formation together with chitinase(s) and beta-glucanase(s) in KA-prep. The N terminal amino acid sequence indicated an analogy of P150T to mutanase (alpha-1,3 glucanase) from Bacillus sp. RM1, and actually P150T hydrolyzed mutan as well as S-(alpha-1,3) glucan from S. commune. PMID- 14519985 TI - Reconstitution and characterization of NtrC protein in a deep-sea piezophilic bacterium, Shewanella violacea strain DSS12. AB - NtrC protein of piezophilic Shewanella violacea was overexpressed and purified, to confirm the protein-DNA interaction. An electrophoretic mobility shift assay demonstrated that the NtrC recognizes the sequence for NtrC binding within the region upstream of the glnA operon. Western blot analysis also showed that the NtrC is expressed at a higher level under high-pressure conditions than under atmospheric pressure conditions. PMID- 14519986 TI - Glycerolipid acyl hydrolase activity in the brown alga Cladosiphon okamuranus TOKIDA. AB - The brown alga, Cladosiphon okamuranus TOKIDA, was found to contain a large amount of free fatty acid (45% of the total lipids). A crude enzyme preparation from the alga showed activity for hydrolyzing the acyl groups of various glycerolipids. The results suggest that the free fatty acid in C. okamuranus was released mainly from glycoglycerolipids, which were the major lipid components in the alga, by such glycerolipid acyl hydrolases as galactolipase. PMID- 14519987 TI - A study of tryptophan fluorescence quenching of bifunctional alginate lyase from a marine bacterium Pseudoalteromonas sp. strain No. 272 by acrylamide. AB - A fluorescence quenching study of a sole tryptophan residue of a bifunctional alginate lyase from Pseudoalteromonas sp. strain No. 272 was done in the presence and absence of substrates, oligomeric guluronic and its C5 isomer mannuronic acid, by a Stern-Volmer plot with a quencher, acrylamide. N-Acetyltryptophanamide and reduced and carboxymethylated alginate lyase showed large quenching constants, on the other hand, the native enzyme had small constants regardless of the presence or absence of the substrates. The result suggests that the tryptophan residue is located in a buried region of the enzyme molecule, but is barely accessible to acrylamide, and that the residue is not masked by the substrates with various degrees of polymerization. PMID- 14519988 TI - Subsite structure of maltogenic amylase from Thermomonospora viridis TF-35. AB - We estimated the subsite structure of the maltogenic amylase from Thermomonospora viridis TF-35 (TVA). TVA has six subsites, and the catalytic site is between the 3rd and 4th subsite. The subsite affinities, A(-3), A(-2), (A(-1)+A(+1)), A(+2) and A(+3) were calculated to be 0.47, 2.31, 0.49, 2.45, and 0.17 kcal mol(-1), respectively. PMID- 14519989 TI - Generation of reactive oxygen species from Hinokitiol under near-UV irradiation. AB - Near-UV irradiation caused the decomposition of hinokitiol in an aqueous solution. During the photochemical reaction, the distinct electron spin resonance signal characteristic of the adduct of 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) with the hydroxyl radical was accompanied by small signals corresponding to the adduct of DMPO with the superoxide anion radical. More than 95% of Escherichia coli cells were killed by the incubation with hinokitiol under near-UV irradiation by BLB fluorescent lamps. These results indicated the generation of reactive oxygen species during photochemical reaction of hinokitiol under near-UV irradiation. PMID- 14519990 TI - (-)-hydroxycitrate ingestion increases fat oxidation during moderate intensity exercise in untrained men. AB - We examined the effects of (-)-Hydroxycitrate (HCA) ingestion on fat oxidation during moderate intensity exercise in untrained men. Six subjects ingested 500 mg of HCA or a placebo for 5 days and did endurance exercise. Blood FFA concentrations were significantly increased and respiratory exchange ratio (RER) decreased by HCA ingestion. These results suggested short-term HCA ingestion increases fat oxidation in untrained men. PMID- 14519991 TI - Nucleotide sequences of genes encoding allosamidin-sensitive and -insensitive chitinases produced by allosamidin-producing Streptomyces. AB - Allosamidin is a strong inhibitor of family 18 chitinases. We previously reported the presence of allosamidin-sensitive and -insensitive chitinases (chitinase S and IS) in the culture filtrate of the allosamidin-producing strain, Streptomyces sp. AJ9463. In this study, we cloned and sequenced the genes encoding the two chitinases, which clarified that chitinase S and IS belong to the family 18 and 19 chitinase, respectively. PMID- 14519992 TI - Construction of a homologous selectable marker gene for Lentinula edodes transformation. AB - We cloned a gene for the iron sulfur protein (Ip) subunit from an edible mushroom, Lentinula edodes, and introduced a point mutation that confers carboxin resistance into it. The mutant gene successfully transformed L. edodes with high efficiency (9 transformants/2.5 microg vector DNA). Restriction enzyme-mediated integration (REMI) increased the transformation efficiency by about two-fold. PMID- 14519993 TI - Cloning and sequence analysis of endoglucanase genes from an industrial fungus, Aspergillus kawachii. AB - Three endoglucanase genes (cel5A, cel5B, and cel61A) were cloned from an industrial fungus, Aspergillus kawachii. Yeasts transformed with these cDNAs showed endoglucanase activity in medium. Cel5A and Cel61A contained a type 1 cellulose-binding domain (CBD1) at the C-terminus of the enzyme. The putative catalytic regions of Cel5A and Cel5B showed homology with various endoglucanases belonging glycosyl hydrolase family 5 (GH5). Cel5B showed high homology with Cel5A in catalytic region, but it lacked CBD1 and linker. The cel5A contained four introns, whereas cel5B contained five introns. The putative catalytic region of Cel61A showed homology with enzymes belonging to GH61. The cel61A contained no introns. PMID- 14519994 TI - Structural effects of phenolic acids on the transepithelial transport of fluorescein in caco-2 cell monolayers. AB - The structural specificity of the monocarboxylic acid transporter (MCT) for the transport of phenolic acids was investigated by measuring the inhibitory effect on the fluorescein transport in Caco-2 cell monolayers. Although most of the monohydroxylated derivatives had an inhibitory effect, the di- and tri hydroxylated ones did not. The methoxylated derivatives were more inhibitory than the hydroxylated ones in all the meta-substituted derivatives, suggesting that meta-hydroxylation of the substrate would decrease the affinity for MCT. PMID- 14519995 TI - Proteases of Maitake (Grifola frondosa) responsible for breakdown of wheat flour dough and their reaction with gluten proteins. AB - Two proteases capable of decreasing dough strength when added to wheat flour were purified from Maitake and these were both thought to be peptidyl-Lys metalloendopeptidase. The major purified protease SP-3-A hydrolyzed high molecular-weight glutenin subunits preferably to the other glutenin subunits. SP 3-A cleaved peptide bonds adjacent to the N-terminal of lysine in the high molecular-weight glutenin subunit. PMID- 14519996 TI - Root growth-promoting activity of unsaturated oligomeric uronates from alginate on carrot and rice plants. AB - The root elongation activity of unsaturated oligomeric uronates from alginate on carrot and rice plants was investigated. Unsaturated oligomeric uronates were prepared by digesting polymannuronate (PM) and polyguluronate (PG) with an alginate lyase purified from Pseudoalteromonas sp. strain No. 272. The root elongation activity was measured by elongation in length of carrot- and rice excised root incubated in the B5-medium containing 0.8% agar in the dark. PM and PG showed no activity, but the enzymatic digestion mixtures of PG had promoting activity on roots of both plants at a final concentration of 0.5 mg/ml. The maximum activity was obtained at 0.75 mg/ml. The dependence of activity on degree of polymerization of the uronates was tested and the pentamer was most active, but the mechanism of the action of unsaturated uronates on the cells remains to be solved. PMID- 14519997 TI - Isolation and characterization of phenol-catabolizing bacteria from a coking plant. AB - New phenol degrading bacteria with high biodegradation activity and high tolerance were isolated as Burkholderia cepacia PW3 and Pseudomonas aeruginosa AT2. Both isolates could grow aerobically on phenol as a sole carbon source even at 3 g/l. The whole-cell kinetic properties for phenol degradation by strains PW3 and AT2 showed a Vmax of 0.321 and 0.253 mg/l/min/(mg protein), respectively. The metabolic pathways for phenol biodegradation in both strains were assigned to the meta-cleavage activity of catechol 2,3-dioxygenase. PMID- 14519998 TI - Fosmidomycin resistance in adenylate cyclase deficient (cya) mutants of Escherichia coli. AB - Adenylate cyclase deficient (cya) mutants of E. coli K-12 were found to be resistant to fosmidomycin, a specific inhibitor of the non-mevalonate pathway, just like to fosfomycin. E. coli glpT mutants were resistant to fosfomycin and also to fosmidomycin. This fact shows that fosmidomycin was transported inside via the glycerol-3-phosphate transporter, GlpT. DNA micro-array analysis showed that the transcription of glpT and other genes concerning glycerol utilization were highly dependent on the presence of cAMP. PMID- 14519999 TI - A simple and eficient method for high fidelity PCR cloning using antibody neutralizing technology. AB - We introduce the TA cloning antibody method for the high-fidelity PCR product amplified by family B DNA polymerase without purification. This method uses antibodies and Thermus aquaticus (Taq) DNA polymerase. The antibodies can inhibit only the activity of family B DNA polymerase, and Taq can co-work for A-tailing. This method has nearly cloning efficiency to that of the PCR product of Taq. PMID- 14520000 TI - Myrsinoic acid E, an anti-inflammatory compound from Myrsine seguinii. AB - The methanolic extract of Myrsine seguinii yielded the novel anti-inflammatory compound, myrsinoic acid E (1), whose structure was elucidated to be 3,5 digeranyl-4-hydroxy benzoic acid. We synthesized 1- and its 3,5-diprenyl (2) and 3,5-difarnesyl analogues (3). Compounds 1-3 suppressed 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced inflammation of mouse ears by 59%, 14%, and 69% at a dose of 1.4 micromol. PMID- 14520001 TI - Activin A induces phosphorylation of Smad2 but not complex formation of Smad2 with Smad4 in human colon cancer cell line HT-29. AB - Western blotting coupled with immunoprecipitation showed that activin A treatment induced phosphorylation of Smad2 but not complex formation of Smad2/4 in human colon cancer-derived HT-29 cells. Because HT-29 cells expressed neither Smad4 mRNA nor Smad4 protein, it is suggested that deletion of Smad4 leads to a defect of formation of Smad2/4 complex upon activin A stimulation in HT-29 cells. PMID- 14520002 TI - Ethyl 4-[2-(6-Methyl-3-pyridyloxy)butyloxy]benzoate, a novel anti-juvenile hormone agent. AB - Ethyl 4-[2-(6-methyl-3-pyridyloxy)butyloxy]benzoate (2) was prepared as a novel anti-juvenile hormone (anti-JH) agent. Compound 2 induced precocious metamorphosis in larvae of the silkworm and black pigmentation of the larval cuticle, which are clearly recognized as JH-deficiency symptoms. The 4 ethoxycarbonyl group on the benzene ring was indispensable for activity. The activity of compound 2 could be fully counteracted by methoprene, a JH agonist, but not by the dietary administration of 20-hydroxyecdysone. PMID- 14520003 TI - A long acidic domain affects the chromatographic behaviour of a neuronal adaptor protein on DEAE-Sepharose. AB - The stepwise chromatographic behaviour on DEAE-Sepharose of rat Fe65, a neuronal protein, was tested, using as eluants KCl, CaCl2, and MgCl2. Assays by western blot showed that Fe65 was eluted by CaCl2, at a ionic strength 20% lower than that of MgCl2 or KCl. Interestingly, in the case of a truncated Fe65, lacking a glutamic acid rich region at the N-terminus, the ionic strengths of the various eluants were almost identical. These results suggested a possible inhibitory role of calcium ions in the binding of the protein to DEAE and a specific affinity of these ions for long acidic stretches. PMID- 14520004 TI - Isolation of lectins by affinity chromatography with porcine plasma proteins immobilized on agarose. AB - To develop a convenient method to isolate lectins, we prepared an affinity gel by coupling plasma proteins with agarose beads under conditions where the pH did not exceed 7.5. The validity of the use of this affinity gel in combination with elution using a hapten saccharide was confirmed by isolation of concanavalin A from Jack bean meal. Successful application of the method was demonstrated by isolation of two novel vegetable lectins from udo (Aralia cordate) and wasabi (Wasabia japonica). The method would be useful to isolate new lectins from various sources including plant and animal tissues. PMID- 14520005 TI - 350-kDa royal jelly glycoprotein (apisin), which stimulates proliferation of human monocytes, bears the beta1-3galactosylated N-glycan: analysis of the N glycosylation site. AB - While doing a structural analysis of minor component N-glycans linked to 350-kDa royal jelly glycoprotein (RJGP), which stimulates the proliferation of human monocytes, we found that a Galbeta1-3GlcNAcbeta1-4Man unit occurs on the insect glycoprotein. The structure of the fluorescence-labeled N-glycan was analyzed by sugar component analysis, IS-MS, and (1)H-NMR. The structural analysis showed that the 350-kDa RJGP bears Galbeta1-3GlcNAcbeta1-4(GlcNAcbeta1-2)Manalpha1-3 (Manalpha1-3Manalpha1-6)Manbeta1-4GlcNAcbeta1-4GlcNAc, suggesting this insect glycoprotein is one of the substrates for both beta1-3 galactosyl and beta1-4 N acetylglucosamininyl transferases. To our knowledge, this is the first report that succeeded in identifying an insect glycoprotein bearing the beta1-3 galactosylated N-glycan. PMID- 14520006 TI - Effect of poly(ADP-ribose) polymerase inhibition on outer medullary hypoxic damage. AB - Poly(ADP-ribose) polymerase (PARP) activation after free-radical-induced DNA damage depletes cellular energy stores and participates in ischemia-reflow injury. We studied the potential protective effect of the water-soluble PARP inhibitor 3-aminobenzamide (3-AB) in a rat model of acute renal failure (ARF) from combined administration of radiocontrast, indomethacin and N(omega)-nitro-L arginine methyl ester. Kidney function at 24 h was better preserved in rats treated with 3-AB as compared to control animals. However, the extent of tubular hypoxic damage was not significantly mitigated. It is concluded that PARP inhibition may attenuate renal dysfunction in this model of ARF with medullary hypoxic tubular injury even while the extent of tubular necrosis is not significantly altered. Further studies of this dyssynchrony of structure and function may provide important insights into the sequence of events that promotes renal failure after medullary injury. PMID- 14520008 TI - Profiling gene expression in kidney development. AB - Insights into the normal development of the kidney have been derived from metanephric organ cultures, knockout mice and the later definition of the responsible single genetic abnormalities in some human congenital renal tract malformations. Deciphering of the human genome and the ability to examine multiple messenger RNA has opened new vistas and the capacity to begin to understand the complex kidney developmental process. In this review we focus on recent large-scale gene expression profiles performed during normal murine and human nephrogenesis and suggest how they might provide insights into normal kidney development as well as aberrations of this normal process that may result in renal dysplasia or pediatric renal malignancy (Wilms' tumor). PMID- 14520007 TI - Extraction of uraemic toxins with activated carbon restores the functional properties of albumin. AB - BACKGROUND: Previous work has demonstrated that a partial normalization of the conformation of albumin from uraemic plasma and a substantial restoration of its binding abilities can be achieved by extraction with activated charcoal. This is best achieved at pH 3, but exposure of whole plasma to this low pH leads to the loss of some essential components. METHODS: The melting curves and ligand-binding abilities of uraemic albumin have been investigated after extraction with a new generation of activated carbon at three pH values (7.2, 3.0 and 5.08). RESULTS: Albumin isolated from uraemic plasma had a characteristically increased melting temperature because of bound ligands. Extraction of uraemic plasma at pH 7.2, 5.08 and 3.0 induced low-temperature shifts of albumin thermo-adsorption maximum T1 of 1.4, 3.8, 2.4 degrees C and T2 of 0.8, 3.9 and 1.2 degrees C, respectively. Flow microcalorimetry data demonstrated a decrease in the ability of uraemic albumin to bind octanoate, phenol red, salicylic acid, warfarin and diazepam. Purification of uraemic plasma at pH 5.08 completely restored the binding affinity of albumin for all the marker ligands. CONCLUSIONS: Highly efficient activated carbons, with clinically feasible acidification of plasma, can remove strongly albumin-bound uraemic toxins. Investigation of the melting curve of the isolated albumin is a new biophysical way to monitor both its molecular condition and the extent of removal of protein-bound toxins by dialysis. The melting curve provides new qualitative and quantitative information about albumin in an analogous way to an electrocardiogram and the heart. PMID- 14520009 TI - Role of Fcgamma receptors in glomerulonephritis. AB - Many forms of glomerulonephritis involve immune complex localization in the kidney. Fcgamma receptors (FcgammaR) expressed on the surface of leukocytes bind the Fc (constant) portion of IgG. They link immune complex deposition to innate immune responses, including phagocytosis, cytokine release, formation of reactive oxygen species, and antibody-dependent cytotoxicity. Activator and inhibitor FcgammaR co-exist on the surface of cells and set thresholds for immune responses. Inhibitor FcgammaR also have a specific role in the maintenance of B cell tolerance. Studies of gene-targeted mice lacking specific FcgammaR have demonstrated that animals lacking activator FcgammaR on circulating leukocytes were protected from immune complex induced glomerular injury, both in models of systemic lupus erythematosus (SLE) and anti-glomerular basement membrane antibody disease. In contrast, mice lacking inhibitor FcgammaR developed SLE-like auto immunity on certain genetic backgrounds and were more susceptible to anti glomerular basement membrane disease, with evidence of enhanced immune responses and dysregulated effector cells. There is also evidence for FcgammaR dysfunction in human SLE and for an association of 'low-binding' polymorphisms of FcgammaR with SLE. In the future, therapeutic possibilities may exist for the manipulation of the FcgammaR pathway in the treatment of glomerulonephritis. PMID- 14520010 TI - Oxidative stress in a novel model of chronic acidosis in LLC-PK1 cells. AB - Chronic metabolic acidosis occurs commonly in chronic renal failure (CRF). The proximal renal tubular cell is the site in the kidney of high oxidative metabolic activity and in CRF is associated with adaptive hypertrophy and hypermetabolism. We hypothesised that chronic acidosis may lead to increased generation of reactive oxygen species due to increased oxidative activity. We developed a novel model of chronic acidosis in LLC-PK1 cells and measured markers of oxidative stress and metabolism. Acidosis led to a reduction in cellular total glutathione and protein thiol content and an increase in glutathione peroxidase activity and NH3 generation. The expression of constitutively expressed heat stress protein (HSP) HSC70 and HSP60 increased at pH 7.0. PMID- 14520011 TI - Role of thyroid hormones on the synthesis and release of atrial natriuretic peptide in rats with acute renal failure. AB - BACKGROUND/AIMS: High atrial natriuretic peptide (ANP) serum levels are seen in acute renal failure (ARF) due to impaired ANP clearance, but the response of ANP to blood volume expansion is blunted. ARF-associated hypothyroidism influences ANP clearance but it may also play a role in the synthesis and release of ANP because in primary hypothyroidism both mechanisms are disturbed. The aim of this study is to analyze whether thyroxin (T4) supplementation improves the synthesis and release of ANP in ARF. METHODS: Four groups of rats were studied: group C (control); group ARF; Group ARF-T4 (ARF supplemented with T4), and group Tx (thyroidectomized). Serum creatinine (SCr), creatinine clearance (CrCl), total T4, total triiodothyronin (T3), plasma ANP, ANP response to acute saline load and atrial content of mRNA-ANP were measured 5 days after ARF induction with potassium dichromate. RESULTS: CrCl was 1.8 +/- 0.4 ml/min in group C, and it significantly dropped to 0.6 +/- 0.5 in ARF (p < 0.01), T4 improved it (1.7 +/- 0.7 in ARF-T4, p < 0.01) and Tx did not change it. Plasma ANP was higher in ARF as compared with C (152 +/- 23 vs. 52 +/- 21 fmol/l, p < 0.01). In ARF-T4, ANP was higher than in C but lower than in ARF (85 +/- 28 fmol/l, p < 0.01). No changes were observed in Tx rats (48 +/- 17 fmol/l). The atrial content of mRNA ANP was 0.06 +/- 0.025 mRNA-ANP/mRNA-beta-actin ratio units in group C, it was higher in ARF (0.13 +/- 0.025, p < 0.01) and lower in Tx (0.04 +/- 0.01, p < 0.05) as compared to C. Supplementation with T4 increased the mRNA-ANF content (0.18 +/- 0.2 mRNA-ANP/mRNA-beta-actin ratio units, p < 0.05) over ARF. Plasma ANP response to saline load was 45 +/- 7% in C, but it was reduced in ARF and Tx (20 +/- 10 and 26 +/- 10%, p < 0.01). In ARF-T4 the response was restored (36 +/- 9%, p < 0.01 vs. ARF). CONCLUSION: Thyroid hormones modulate the synthesis and release of ANP in ARF. Although the effect is probably direct, T4-induced amelioration of renal damage may also play a significant role by improving the uremic milieu. PMID- 14520012 TI - Angiotensin-converting enzyme inhibition modulates high-glucose-induced extracellular matrix changes in mouse glomerular epithelial cells. AB - BACKGROUND/AIM: Extracellular matrix alterations are involved in the pathogenesis of diabetic nephropathy. We evaluated the effects of high glucose concentrations and inhibition of angiotensin-converting enzyme on the laminin and fibronectin production by glomerular epithelial cells. METHODS: Glomerular epithelial cells were cultured in 5 and 30 mmol/l glucose, with and without enalaprilat (0.3 mmol/l). Laminin and fibronectin were measured (35S-methionine, immunoprecipitation), and their mRNA expression was evaluated (RT-PCR). RESULTS: The laminin concentration was higher in the cells than in the medium, where an increase of its content was observed under high-glucose conditions (p < 0.01). Fibronectin, found only in the medium, was not modified by the high glucose concentration. Following enalaprilat administration, the laminin concentration was decreased under high-glucose conditions, both in the cell and in the medium (p < 0.001), whereas the fibronectin concentration was increased under high glucose conditions (p < 0.001). The mRNA expression of laminin and fibronectin under high-glucose conditions only slightly increased. Enalaprilat decreased the fibronectin mRNA synthesis dramatically (>50%, p < 0.0001) under high-glucose conditions. CONCLUSIONS: Enalaprilat normalizes the abnormal, high-glucose induced concentration of laminin, while it decreases the fibronectin synthesis. The improvement of the renal function in diabetic patients treated with angiotensin-converting enzyme inhibitors may, in part, be due to a modulator effect on extracellular matrix content and composition. PMID- 14520013 TI - Involvement of p300 in TGF-beta/Smad-pathway-mediated alpha2(I) collagen expression in mouse mesangial cells. AB - BACKGROUND: Transforming growth factor beta 1 (TGF-beta1) induces alpha2(I) collagen gene (COL1A2) expression in mesangial cells through physical and functional cooperation of Smad proteins and Sp1. A transcriptional coactivator, p300, is also suggested to play an important role in TGF-beta1/Smad signal transduction. However, the role of p300 in TGF-beta1/Smad-pathway-mediated transcriptional activation of the COL1A2 gene in mesangial cells is still obscure. METHODS: Endogenous p300 expression and its modulation by TGF-beta1 were evaluated by Western blotting and immunofluorescence. The physical interaction of p300 with Smad2/3 was examined by immunoprecipitation followed by Western blotting. The functional role of p300 in TGF-beta1/Smad-pathway-mediated COL1A2 transcription was investigated in cotransfection experiments using a COL1A2 promoter-luciferase reporter gene construct and p300 expression plasmids. RESULTS: TGF-beta1 induced COL1A2 gene expression in cultured mouse mesangial cells which was blocked by overexpression of inhibitory Smad7. In addition, TGF beta1-induced nuclear export of endogenous Smad7 was observed in mouse mesangial cells. Endogenous p300 was expressed in the nucleus of the cells. TGF-beta1 induced interaction of endogenous p300 with Smad2/3, and a dominant negative construct of p300 inhibited the TGF-beta1-induced COL1A2 expression in cultured mouse mesangial cells. CONCLUSIONS: p300 may be involved in TGF-beta1/Smad pathway-mediated type I collagen gene transcription in mouse mesangial cells. Our findings would reveal a molecular basis of TGF-beta1-induced type I collagen gene transcription in mouse mesangial cells. PMID- 14520014 TI - Examine the individual, not the species... PMID- 14520015 TI - Complement in renal transplantation. AB - Previous research and therapy in renal transplantation largely focused on the cellular arm of the adaptive immune response. Evidence is emerging that innate immune mechanisms, particularly complement, play a greater role in inflammatory and immune responses against the graft than has been previously recognized. Alternative complement pathway activation appears to mediate renal ischaemia/reperfusion injury, and proximal tubular cells may be both the source and the site of attack of complement components in this setting. Locally produced complement also plays a role in the development of both cellular and antibody mediated immune responses against the graft. C4d staining has emerged as a useful marker of humoral rejection both in the acute and in the chronic setting and led to renewed interest in the significance of anti-donor antibody formation. A number of therapies are in development which inhibit complement or reduce local synthesis, and may lead to an improved clinical outcome following renal transplantation. PMID- 14520016 TI - Lean mass estimation by creatinine kinetics and dual-energy x-ray absorptiometry in peritoneal dialysis. AB - BACKGROUND: Malnutrition is widely prevalent in dialysis. Malnourished patients present depletion of somatic protein stores and a decrease in lean body mass (LBM) that can be measured by different techniques. AIMS: (1) To assess the reliability of lean mass measurements obtained by creatinine kinetics (CrK) in a group of stable peritoneal dialysis (PD) patients, using dual-energy x-ray absorptiometry (DEXA) measurements as the reference method; (2) to establish the reproducibility of LBM estimated by CrK in individual patients analyzing repeated measurement in the short term, and (3) to correlate measurements of LBM with laboratory determinations that assess nutritional status. METHODS: We performed a cross-sectional evaluation of LBM by DEXA and CrK in 39 PD patients. In 14 patients we performed repeated measurements of LBM by CrK in the short term. RESULTS: No significant difference was found in mean lean mass values estimated by both methods: mean DEXA LBM was 41.7 kg, 36.1 +/- 4.5 kg in females and 52.7 +/- 6.4 kg in males and mean CrK LBM was 41.08 kg, 37.5 +/- 6.1 kg in females and 48.1 +/- 8.4 kg in males. A good correlation was found between both techniques (r = 0.71; p < 0.003). The mean difference between the two methods was 0.638 +/- 6.95 kg (95% confidence limits: -12.98 and +14.26). A wide scatter of the differences between both methods was seen throughout the range of measurements of LBM. When LBM by CrK was repeatedly (2-3 times) measured in a period of 3-4 months in 14 patients, it had a coefficient of variation (CV) of 15.39% (range 2.89-42.88%), while body weight CV in the same period was 0.69% (range 0-1.9%). CONCLUSIONS: CrK is an unsatisfactory method for the assessment of LBM in PD patients. PMID- 14520017 TI - Predictors of recurrent hyperparathyroidism after total parathyroidectomy in chronic renal failure. AB - BACKGROUND/AIMS: Recurrent hyperparathyroidism (HPT) after total parathyroidectomy (TPTX) in chronic renal failure appears more common than might be anticipated. METHODS: To study its predictors, we reviewed all 20 cases of TPTX performed at our hospital in a 10-year period. RESULTS: During follow-up (median 46.8 months (range 9.3-120.3)), 15 patients had measurable PTH levels (>10 pg/ml), 7 had levels above the normal range (recurrent HPT), and 3 had PTH levels >300 pg/ml (severe recurrent HPT). Total follow-up post-TPTX was equal in those who developed recurrent HPT and others, but those with recurrent HPT had spent longer on dialysis post-TPTX (61.9 +/- 34.9 vs. 21.8 +/- 12.0 months; p = 0.001). Patients with recurrent HPT required less vitamin D supplementation during the 10 days post-TPTX (p = 0.025). Log [maximal PTH post-TPTX] correlated with duration of dialysis dependency post-TPTX (r = 0.591, p = 0.006), lowest serum calcium level during the first 30 days post-TPTX (r = 0.449, p = 0.047), and mean serum calcium during the first 30 days post-TPTX (r = 0.546, p = 0.013). Mean log [maximal PTH post-TPTX] was significantly lower in patients with ectopic calcification (p = 0.047). In multiple regression analysis, duration of dialysis post-TPTX and lowest serum calcium level during the first 30 days post-TPTX were the only independent predictors of log [maximal PTH post-TPTX]. CONCLUSION: Recurrent HPT is common following TPTX and predicted by duration of dialysis dependency post-TPTX, a measure of overall exposure to the uraemic stimulus to parathyroid hyperplasia, and the degree of early hypocalcaemia, possibly reflecting the adequacy of operative parathyroid ablation. PMID- 14520018 TI - Clinical effect of short daily in-center hemodialysis. AB - BACKGROUND: The safety and efficacy of short daily hemodialysis has been reported in the USA and Europe, but there is no report on its efficacy in Japanese patients undergoing hemodialysis. METHODS: Twenty-three outpatients (14 men and 9 women, 55.8 +/- 9.6 years old and 11.1 +/- 6.6 years on dialysis) undergoing hemodialysis 3 times/week participated in this study. After 4 weeks' baseline observation under conventional hemodialysis, they were subjected to short daily in-center hemodialysis (DHD, 6 times/week) for 12 weeks and then a 4-week follow up observation period under conventional hemodialysis. RESULTS: The mean pre dialysis systolic and diastolic blood pressure significantly decreased in the DHD period. Antihypertensive drugs could be discontinued or the dose was reduced in 6 of 11 patients treated with such drugs. The hematocrit level tended to increase in the DHD period, and recombinant human erythropoietin could be discontinued or reduced in 7 of 14 patients. Localized skin rash caused by the adhesive tape and lidocaine patch at the blood access was observed in only 2 patients, but no other adverse events associated with DHD were noted. CONCLUSION: These results indicate that DHD is safe and more useful than conventional 3 times/week hemodialysis for Japanese patients undergoing hemodialysis. PMID- 14520019 TI - Influence of glucose in dialyzing fluid on purine concentrations in hemodialyzed patients with chronic renal failure. AB - BACKGROUND: In chronic renal failure the accumulation of some purine nucleotides (in erythrocytes) develops both in patients undergoing conservative treatment and in hemodialyzed patients. The aim of the study was: (1) To find if hemodialysis (HD) sessions using dialyzing fluid containing glucose leads to an increase in ATP concentration and changes in the concentration of other nucleotides, nucleosides and oxypurines in erythrocytes. The potential consequence of such purine concentration changes is the increase of 2,3-DPG concentration and an improved transportation of oxygen in erythrocytes which are more resistant to hemolysis. (2) To compare blood concentrations of purine nucleotides, nucleosides and oxypurines in patients undergoing chronic HD with dialyzing fluid containing or lacking glucose. Significant differences could suggest the long-term influence of glucose in dialyzing fluid on erythrocyte energetic state. METHODS: Whole blood nucleotide concentrations were evaluated with the use of a high-performance liquid chromatography technique. RESULTS: Before the HD session the patients in the 'plus glucose' group had significantly higher concentrations of ATP, ADP, AMP, TAN, NAD, NADP, GTP + GDP, GMP, Urd and HYP than patients in the 'no glucose' group. After the HD the patients in the 'plus glucose' group had significantly higher concentrations of ADP, AMP, TAN, NAD, NADP, Urd and HYP than in the 'no glucose' group. Both before and after the HD session, the uric acid concentrations and AEC were significantly lower in the 'plus glucose' group than in the 'no glucose' group. A significant decrease in the whole blood hypoxanthine (p < 0.05) and uric acid (p < 0.001) concentrations after HD was found in the 'no glucose' group while a significant increase in ADP concentration (p < 0.05) was detected in the patients' erythrocytes in the 'plus glucose' group. In this group a significant decrease of GTP + GDP and GMP (p < 0.05), uric acid concentration (p < 0.001) and adenylate energy charge (p < 0.05) were observed after the dialysis. However, no significant differences in nucleotide concentrations before and after the HD were found in the 'no glucose' group. CONCLUSION: The presence of glucose in the dialyzing fluid causes a significant modification of the energetic state of cells which is reflected by the purines' and their metabolites' concentrations in the erythrocytes. Higher ATP concentrations in patients with renal failure who have been dialyzed with the fluid containing glucose can be considered as an organism adaptation to a decreased amount of RBC and hemoglobin concentration. PMID- 14520020 TI - Supplementation of a low-carotenoid diet with tomato or carrot juice modulates immune functions in healthy men. AB - BACKGROUND: Beta-carotene has been shown to enhance immune functions in humans. Whether vegetables rich in carotenoids, such as beta-carotene or lycopene, modulate immune functions in healthy humans is presently not known. The objective of this study was to investigate the effects of a low-carotenoid diet supplemented with either tomato (providing high amounts of lycopene) or carrot juice (providing high amounts of alpha- and beta-carotene) on immune functions in healthy men. METHOD: In a blinded, randomized, cross-over study, male subjects on a low-carotenoid diet consumed 330 ml/day of either tomato juice (37.0 mg/day lycopene) or carrot juice (27.1 mg/day beta-carotene and 13.1 mg/day alpha carotene) for 2 weeks with a 2-week depletion period after juice intervention. Immune status was assessed by measuring lytic activity of natural killer (NK) cells, secretion of cytokines (IL-2, IL-4, TNFalpha), and proliferation by activated peripheral blood mononuclear cells. RESULTS: Juice consumption resulted in relatively fast responses in plasma carotenoid concentrations (p < 0.0002) which were not accompanied by concomitant changes in immune functions. For IL-2, NK cell cytotoxicity, and lymphocyte proliferation, maximum responses were observed during depletion periods. The highest production rate was measured only for TNFalpha at the end of the first intervention period. Juice intervention did not modulate the secretion of IL-4. CONCLUSIONS: Increased plasma carotenoid concentrations after vegetable juice consumption are accompanied by a time delayed modulation of immune functions in healthy men consuming a low-carotenoid diet. PMID- 14520021 TI - The role of dietary supplementation with L-glutamine in inflammatory mediator release and intestinal injury in hypoxia/reoxygenation-induced experimental necrotizing enterocolitis. AB - BACKGROUND/AIMS: Necrotizing enterocolitis (NEC) is a multifactorial syndrome in the neonate. Enteral feeding practices are an important component of gastrointestinal injury in neonatal NEC. In the present study, we examined the protective effect of oral supplementation with L-glutamine, an important specific fuel for the enterocytes, against hypoxia-reoxygenation (H/R)-induced NEC in young mice. METHODS: Young mice were divided into four groups: group 1 mice (untreated) underwent H/R; group 2 mice were supplemented with L-glutamine in drinking water (0.5 g/dl) for 3 days, and group 3 mice were supplemented with L glutamine (3 g/dl) for 10 days. Group 4 mice served as control. Hypoxia was induced by placing the young mice in a 100% CO(2) chamber for 5 min. After hypoxia, they were reoxygenated for 10 min with 100% oxygen. We examined the intestinal lesions with light microscopy and measured intestinal generation of PAF and TNF-alpha in the H/R-induced model of NEC. RESULTS: In group 3 mice, NEC induced intestinal tissue damage was greatly attenuated with necrosis limited partially to the mucosa. Both intestinal tissue PAF and TNF-alpha concentrations were significantly higher in the untreated group than in controls (p < 0.001). Group 3 mice (3 g/dl supplemented) showed a significant decrease in intestinal TNF-alpha concentration compared with young group 1 and group 2 mice (p < 0.05 and p < 0.05, respectively). On the other hand, no significant difference was observed in the intestinal generation of PAF between H/R groups (p > 0.05). CONCLUSION: The present study suggests that H/R plays an important role in the pathogenesis of NEC and supports the hypothesis that especially PAF and TNF-alpha are involved in the pathophysiological mechanism of H/R-induced NEC. This study also demonstrates that dietary supplementation with L-glutamine reduces the histologic evidence of H/R-induced intestinal injury. Based on these findings, beneficial effects of L-glutamine in this model of NEC are mediated via mechanisms inhibiting intestinal cytokine release. PMID- 14520022 TI - Dietary intake of 10- to 16-year-old children and adolescents in central and northern Europe and association with the incidence of type 1 diabetes. AB - BACKGROUND/AIMS: The highest incidence of type 1 diabetes is among 10- to 15-year old adolescents. The aim of this study was to investigate a possible relationship between the dietary intake of this group and the incidence of type 1 diabetes. METHODS: Dietary intake data of 10- to 16-year-old adolescents (n = 4,701) from 11 European countries and the incidence rates of type 1 diabetes were used to examine the relation between food and the disease. RESULTS: The incidence of type 1 diabetes correlated with the consumption of total fat (r = 0.674; p = 0.023), saturated fatty acids (r = 0.714; p = 0.047) and the intake of fruits and vegetables (r = 0.786; p = 0.036). Fruit intake or vegetable intake alone did not correlate with the incidence. Cow's milk and animal product consumption correlated with the incidence when Icelandic data were excluded (r = 0.829; p = 0.042 and r = 0.999; p = 0.001). A negative correlation of borderline significance was found between sugar intake and the incidence of type 1 diabetes (r = -0.721; p = 0.068). CONCLUSION: The results indicate for the first time that an adolescent's diet high in fat and fruits and vegetables is associated with an increased risk of type 1 diabetes. Fruit or vegetable intake separately was not associated with type 1 diabetes. It is important to characterize and minimize diabetogenic factors in fruits and vegetables as the general health benefits of a diet rich in fruits and vegetables are well known and such a diet is therefore recommended. This study supports previous research about the importance of cow's milk and animal products in the aetiology of type 1 diabetes. PMID- 14520023 TI - Serum retinol, iron status and body composition of lactating women in Nandi, Kenya. AB - BACKGROUND: Maternal vitamin A and iron status was investigated among lactating mothers in a rural community in Kenya. The aim of the study was to establish the prevalence and the relationship of these key nutrients to maternal body composition. METHODS: Eighty-eight mothers provided samples of breast milk and blood for determination of breast milk vitamin A, serum retinol and ferritin. Estimators of body composition were based on the mother's weight, height and skinfold measurements. RESULTS: A total of 78.1% women had breast milk retinol <1.05 micromol/l with 38 and 62% having lactated for a period of <4 and >4 months, respectively. Prevalence of severely deficient serum retinol <0.35 micromol/l and ferritin <12 microg/l was 10 and 37%, respectively. Women with serum ferritin <12 microg/l had significantly lower average hemoglobin (p < 0.01), hematocrit (p < 0.01) and serum retinol (p < 0.05). Serum retinol of mothers who had lactated for <4 months was significantly but negatively correlated with total body fat (r = -0.40; p < 0.05). With a lactation period of >4 months a close relationship was found between serum retinol and hemoglobin (r = 0.26; p <0.01), serum retinol and serum ferritin (r = 0.20; p < 0.05), and fat free mass significantly but negatively correlated with breast milk fat (r = 0.27; p < 0.05). Serum retinol in combination with hematocrit significantly affected both maternal hemoglobin (p < 0.01) and serum ferritin (p < 0.01). CONCLUSION: A high prevalence of vitamin A and iron deficiency was observed in this group of lactating women. Low levels of fat mass were directly related to these indicators of malnutrition. PMID- 14520024 TI - Body composition, eating behavior, food-body concerns and eating disorders in adolescent girls. AB - AIMS: Dieting is a behavioral phenomenon which is becoming more frequent among adolescents and the search for weight loss, through dieting, may result in an unbalanced nutrition both quantitatively and qualitatively. Our study intended to look at the eating habits and behavior on a cohort of adolescent girls to verify the presence of unbalanced diets and the prevalence of eating disorders with particular attention to the partial syndromes (EDNOS). METHODS: A cross-sectional double-stage study was carried out on a group of schoolgirls in the suburbs of Naples. We assessed anthropometrical measures, body composition (skinfolds and bioimpedance analysis), dietary intake by means of 3-day food records and we administered the Eating Disorder Inventory 2 and Psychosocial Factor Risk Questionnaire. A multidisciplinary and double-stage approach had been used to get a better diagnosis of eating disorders in our sample. RESULTS: 156 adolescent girls, 14-18 years old, took part in our study. Height, weight, and BMI were 160.38 cm, 58 kg and 22.6, respectively. Analysis of food intake showed that all the values reported, with the exception of lipids and sodium, were below the recommendations by LARN. We observed a prevalence of 1.28% of bulimia nervosa, 1.28% of binge eating, and 10.25% of eating disorders not otherwise specified. EDI 2 and PRFQ confirmed how important drive for thinness and body dissatisfaction dimensions are when we deal with adolescent girls and with the phenomenon of dieting. The study confirmed the validity of the PRFQ questionnaire to evaluate mass media influence on body perception and eating behavior of adolescents. CONCLUSION: Multidisciplinary and well-designed studies are needed to systematically and accurately study eating habits and behavior of adolescents to tackle more efficiently the increasing spread of eating disorders and obesity. PMID- 14520025 TI - Effect of vitamin C supplementation on oxidative DNA damage in an experimental model of lead-induced hypertension. AB - AIMS: Chronic exposure to lead results in sustained hypertension in humans and experimental animals. We investigated the possible role of reactive oxygen species (ROS) and their impact on DNA damage in lead-induced hypertension. Further the effect of short-term supplementation of vitamin C is also demonstrated. METHODS: Male Wistar rats were treated with either lead acetate (100 ppm) alone or lead acetate plus vitamin C (20 mg/rat/day). The control rats were fed regular rat chow. Blood pressure, antioxidants, total antioxidant status as measured by ferric-reducing antioxidant power, nitric oxide (NO) metabolites, malondialdehyde (MDA) and 8-hydroxy 2-deoxyguanosine were determined after 0, 1, 2 and 3 months. RESULTS: The lead-exposed group showed a significant rise in blood pressure, lipid peroxidation (MDA) and a substantial oxidative damage to the DNA. A significant fall in NO metabolites, total antioxidant levels and ferric-reducing antioxidant power was also observed in this group. Concomitant administration of vitamin C ameliorated hypertension, normalized NO levels and abrogated lipid peroxidation. Also, it completely prevented oxidative damage to the DNA. CONCLUSIONS: These findings point to enhanced ROS-mediated inactivation and sequestration of NO which can potentially contribute to hypertension, lipid peroxidation, reduced antioxidant status and oxidative DNA damage. The beneficial effects of vitamin C on these parameters support the role of increased ROS activity in the pathogenesis of these abnormalities in this model. PMID- 14520026 TI - Higher content of 18:1 trans fatty acids in subcutaneous fat of persons with coronarographically documented atherosclerosis of the coronary arteries. AB - AIM: To identify the total content of trans fatty acid (TFA) isomers and C18:1 trans isomers in subcutaneous fat samples from persons with atherosclerosis of the coronary arteries, as an indicator of dietary exposure. METHODS: Using capillary gas chromatography, the authors determined total content of TFA isomers and C18:1 trans isomers in the subcutaneous fat of 34 patients with ischemic heart disease who had undergone aortocoronary bypass surgery and in 46 patients with no sign of coronary disease. RESULTS: On average, the total TFAs in cardiac patients were 2.88 +/- 1.19% of all fatty acids, in noncardiac patients 2.56 +/- 0.89%. However, the difference is not statistically significant. The average concentration of C18:1 trans in cardiac patients (2.31 +/- 1.09%) was statistically significantly higher (p = 0.05) than in the noncardiac group (1.95 +/- 0.77%). CONCLUSIONS: The results obtained indicate a lower TFA load in comparison with previous studies in other countries. A higher concentration of 18:1 TFAs in the subcutaneous fat of patients with coronary disease might be an impulse to correct the dietary habits of this very high-risk population. PMID- 14520027 TI - Effect of an oat bran enriched diet on the atherogenic lipid profile in patients with an increased coronary heart disease risk. A controlled randomized lifestyle intervention study. AB - AIM: To study whether an oat bran enriched diet has a specific effect in lowering total and low-density lipoprotein cholesterols, in addition to caloric and fat restriction. METHODS: We performed a randomized, controlled, parallel-group, single-centre study in which 1,994 patients from the Wehrawald Hospital (Todtmoos, Germany) were screened and 235 met the criteria male gender, hypercholesterolemia, and overweight. All patients in the Wehrawald Hospital took part in a 4-week standardized inpatient lifestyle health program consisting of dietary intervention, increased physical activity, and health education. Caloric restriction, fat modification, and oat bran supplementation were part of the nutritional regimen within the lifestyle health program. Ninety-nine patients were randomized to a fat-modified diet with caloric restriction and a daily intake of 35-50 g oat bran and 136 patients to a fat-modified, oat bran-free diet with caloric restriction. Fifty-three male overweight but normocholesterolemic subjects were selected as controls. RESULTS: The most significant decreases in total cholesterol (-67.7 +/- 37.2 mg/dl; p < 0.01), low-density lipoprotein cholesterol (-56.3 +/- 35.1 mg/dl; p < 0.01), and apolipoprotein B (-42.4 +/- 34.1 mg/dl; p < 0.01) were found with the combination of the fat-modified and oat bran enriched food. CONCLUSIONS: Added to a fat-modified diet, oat bran within a practical range of intake significantly reduces total cholesterol, low-density lipoprotein cholesterol, and apolipoprotein B. These beneficial effects occurred independent of covariables such as physical activity or caloric and fat restriction in the diet. PMID- 14520028 TI - Endocrine effects of food intake: insulin, ghrelin, and leptin responses to a single bolus of essential amino acids in humans. AB - BACKGROUND: This study deals with the physiological effects of an oral bolus of essential amino acids (AA) on the secretion of insulin, ghrelin and leptin in fasting humans. METHODS: 12 healthy adults (age 18-40 years, 10 males, 2 females, body mass index 18.0-23.5 kg/m(2)) were included in our study. Seven fasted volunteers consumed an AA mixture (0.35 g/kg BW), 5 served as controls. Serum AA, glucose, albumin, urea and hormones were measured at 0, 15, 30 min and thereafter at 30-min intervals up to 5 h. RESULTS: Glucose, albumin and urea remained constant, peak AA concentrations were achieved at 30 min for methionine (Met), at 60 min for all other AA. The insulin peak (533% compared to basal level, p < 0.01) at 30 min was earlier than the peaks of all AA except for Met. Ghrelin showed a continuous rise towards the end of the experiment leading to a 3-fold increase in initial concentrations in the study group (p < 0.001), significantly higher than in the control group (p < 0.05). In contrast, both groups exhibited almost constant leptin concentrations. CONCLUSION: Our data indicate that an oral low-dose AA bolus not only causes considerable hyperaminoacidemia and hyperinsulinemia but is also accompanied by an increased ghrelin secretion in fasted humans. This may be a specific effect or may be due to the fact that a single bolus of AA is not a sufficient stimulus to induce satiety. Leptin, however, is not a key mediator in this setting. PMID- 14520029 TI - A randomized trial of combined manipulation, stabilizing exercises, and physician consultation compared to physician consultation alone for chronic low back pain. AB - STUDY DESIGN: A prospective randomized controlled trial. OBJECTIVES: To examine the effectiveness of combined manipulative treatment, stabilizing exercises, and physician consultation compared with physician consultation alone for chronic low back pain. SUMMARY OF BACKGROUND DATA: Strong evidence exists that manual therapy provides more effective short-term pain relief than does placebo treatment in the management of chronic low back pain. The evidence for long-term effect is lacking. METHODS: Two hundred four chronic low back pain patients, whose Oswestry disability index was at least 16%, were randomly assigned to either a manipulative-treatment group or a consultation group. All were clinically examined, informed about their back pain, provided with an educational booklet, and were given specific instructions based on the clinical evaluation. The treatment included four sessions of manipulation and stabilizing exercises aiming to correct the lumbopelvic rhythm. Questionnaires inquired about pain intensity, self-rated disability, mental depression, health-related quality of life, health care costs, and production costs. RESULTS: At the baseline, the groups were comparable, except for the percentage of employees (P = 0.01). At the 5- and 12 month follow-ups, the manipulative-treatment group showed more significant reductions in pain intensity (P < 0.001) and in self-rated disability (P = 0.002) than the consultation group. However, we detected no significant difference between the groups in health-related quality of life or in costs. CONCLUSIONS: The manipulative treatment with stabilizing exercises was more effective in reducing pain intensity and disability than the physician consultation alone. The present study showed that short, specific treatment programs with proper patient information may alter the course of chronic low back pain. PMID- 14520030 TI - The effects of an interspinous implant on the kinematics of the instrumented and adjacent levels in the lumbar spine. AB - STUDY DESIGN: Measurement of the kinematics of the lumbar spine after insertion of an interspinous spacer in vitro. OBJECTIVES: To understand the kinematics of the instrumented and adjacent levels due to the insertion of this interspinous implant. SUMMARY OF BACKGROUND DATA: An interspinous spacer (X Stop, SFMT, Concord, California) has been developed to treat neurogenic intermittent claudication by placing the stenotic segment in slight flexion and preventing extension. This restriction of motion by the interspinous implant may affect the kinematics of levels adjacent to the instrumented level. METHODS: Seven lumbar spines (L2-L5) were tested in flexion-extension, lateral bending, and axial rotation. Images were taken during each test to determine the kinematics of each motion segment. The interspinous implant was placed at the L3-L4 level, and the test protocol was repeated. RESULTS: The flexion-extension range of motion was significantly reduced at the instrumented level. Axial rotation and lateral bending ranges of motion were not affected at the instrumented level. The range of motion in flexion-extension, axial rotation, and lateral bending at the adjacent segments was not significantly affected by the implant. CONCLUSIONS: The implant does not significantly alter the kinematics of the motion segments adjacent to the instrumented level. PMID- 14520031 TI - Experimental scoliosis in an immature goat model: a method that creates idiopathic-type deformity with minimal violation of the spinal elements along the curve. AB - STUDY DESIGN: Experimental scoliosis was created in an immature goat model. OBJECTIVES: To create a progressive, structural, idiopathic-type, lordoscoliotic curve convex to the right in the thoracic spine of the immature goat while maintaining the anterior and posterior elements of the spine along the maximal curve in a pristine state. SUMMARY OF BACKGROUND DATA: Progressive scoliotic curves in a small animal model have been created using a posterior asymmetric tether. However, attempts in larger animal models have had less success and typically required violation of the spinal elements. METHODS: Scoliosis was created in 40 Spanish Cross X female goats (age 1-2 months, weight 8-12 kg) using a left posterior asymmetric tether from T5 to L1. Convex rib resection and concave rib tethering from T8 to T13 were performed before compression across the rigid construct. Goats were followed over a 6- to 15-week period with serial radiographs to document progression of the deformity. RESULTS: Of the 40 goats that underwent posterior assymetric tethering with rib procedures, 7 (18%) encountered substantial complications (five deaths and two neurologic injuries). Of the 33 available for analysis, 27 goats (82%) developed progressive, structural, idiopathic-type, lordoscoliotic curves in convex to the right in the thoracic spine. All curves demonstrated characteristic radiographic features of idiopathic scoliosis including significant displacement of the apical vertebra from the midline, wedging of both the vertebral bodies and discs, rotation, and decreased flexibility. Initial scoliosis after posterior asymmetric tethering measured 42 degrees on average (range 33-50 degrees ) and progressed to 60 degrees on average (range 44-73 degrees ) over 6 to 15 weeks. The average progression of +18 degrees (range 6-37 degrees ) was statistically significant (P < 0.001). CONCLUSIONS: This study establishes an experimental model for scoliosis that creates progressive, structural, idiopathic-type, lordoscoliotic curves convex to the right in the thoracic spine of the immature goat with high statistical significance (P < 0.001). In addition, this method of experimental scoliosis creation avoids violation of the spinal elements throughout the maximal portion of the curve providing an ideal opportunity subsequent study of the deformity. PMID- 14520032 TI - Failure of human cervical endplates: a cadaveric experimental model. AB - STUDY DESIGN: An in vitro biomechanical study using a servohydraulic testing machine on cadaveric endplates. OBJECTIVES: To characterize the effects of bone mineral density, endplate geometry, and preparation technique on endplate failure load. SUMMARY OF BACKGROUND DATA: The effects of endplate preparation methods on failure loads are only partly characterized in the literature. Endplate burring has been recommended to increase fusion rates. However, graft subsidence may complicate anterior reconstruction procedures. METHODS: After radiographic screening, 21 cadaveric cervical spines underwent dual-energy x-ray absorptiometry scanning to quantify mineral content. Endplate geometry was calculated in 55 randomly selected endplates from the inferior C2 to the superior T1 levels. These vertebrae were embedded in polyester resin and randomly left intact, perforated, or burred. The cervical endplates were loaded at a rate of 0.2 mm/s on an Instron materials tester with an attached 9 mm diameter polycarbonate rod (an area of 64 mm2). A stepwise, univariate linear regression was used to compare the point of endplate failure with the vertebral level, endplate area, gender, age, bone mineral density, and preparation technique. RESULTS: Mean bone mineral density, as measured by dual-energy x-ray absorptiometry, was 0.713 g/cm2 (+/- 0.173 g/cm2). Mean endplate area was calculated at 323 mm2. A mean compressive force of 754 N (+/- 445 N) was required before endplate failure. Trends toward increasing compressive loads were noted with decreasing endplate area and increasing bone mineral density. Increasing age (P = 0.0203), caudal vertebral level (P < 0.0001), endplate burring (P = 0.0068), and female gender (P = 0.0452) were associated with significantly lower endplate fracture loads in compression. CONCLUSIONS: Bone quality was predictive of endplate compressive failure loads. Intact endplates failed at significantly higher loads than their perforated or burred counterparts. PMID- 14520033 TI - Pathomechanism of myelopathy and surgical results of laminoplasty in elderly patients with cervical spondylosis. AB - STUDY DESIGN: Clinical and radiologic analyses in elderly patients with cervical myelopathy. OBJECTIVE: To investigate the pathomechanism of cervical myelopathy and the surgical results of laminoplasty in elderly patients older than 70 years old. SUMMARY OF BACKGROUND DATA: To date, there have been no reports on the pathomechanism of cervical myelopathy in elderly patients. Further, the surgical results and postoperative complications of laminoplasty have not been fully evaluated in elderly patients. METHODS: Eighty-nine patients who underwent cervical laminoplasty were reviewed. The patients were divided into 2 groups according to the age at the time of operation (the elderly patient group: 20 patients who were older than 70 years old, and the control group: 69 patients who were younger than 69 years old). Pre- and postoperative neurologic status (the Japanese Orthopedic Association score) and postoperative complications were compared between the two groups. Radiologic features were also examined. RESULTS: There was no statistical difference in the recovery rate of Japanese Orthopedic Association score between the elderly patient group and the control group. Activities of daily living improved in the elderly patients. Several complications, such as delirium and worsening hypertension, were found in the elderly patient group. In the preoperative radiographs, the incidence of either retrolisthesis or anterolisthesis in the elderly patient group was significantly higher than that in the control group. CONCLUSIONS: Retrolisthesis and anterolisthesis are often the cause of myelopathy in elderly patients. Surgical decompression for cervical myelopathy was beneficial even in elderly patients older than 70 years old. Laminoplasty achieves stability of the cervical spine, and this procedure is reasonable for the treatment. PMID- 14520035 TI - Reliability of clinical tests in the assessment of patients with neck/shoulder problems-impact of history. AB - STUDY DESIGN: A clinical trial on patients receiving neck/shoulder physical examinations. OBJECTIVES: To analyze reliability of clinical tests, prevalence of positive findings in the assessment of neck/shoulder problems in primary care patients, and the impact of history, including pain drawing, on these parameters. SUMMARY OF BACKGROUND DATA: Reliability of clinical tests varies, perhaps partly because of the impact of history. To our knowledge, this has not been studied before. METHODS: Two examiners independently assessed 100 patients with a set of 66 clinical tests divided into 9 categories. Half of the patients were examined with and the other half without knowledge of history. Reliability as expressed by percentage agreement, kappa coefficients, and prevalence of positive findings was calculated. RESULTS: Reliability of clinical tests was poor or fair in several categories and did not alter with history. Only a bimanual sensitivity test reached good kappa values. With known history, prevalence of positive findings increased. Bias was apparent in all test categories except sensitivity tests. Four out of five patients were diagnosed to have neurogenic dysfunction in the affected area. CONCLUSIONS: Our sensitivity test was the most reliable and also exempt from bias and should be studied further. Some common tests may not be reliable. History had no impact on reliability of our tests but increased the prevalence of positive findings. Neurogenic dysfunction seems very common in patients with neck and/or shoulder problems and should be screened for. PMID- 14520034 TI - Increased sagittal plane segmental motion in the lower cervical spine in women with chronic whiplash-associated disorders, grades I-II: a case-control study using a new measurement protocol. AB - STUDY DESIGN: Case-control study comparing sagittal plane segmental motion in women (n = 34) with chronic whiplash-associated disorders, Grades I-II, with women (n = 35) with chronic insidious onset neck pain and with a normal database of sagittal plane rotational and translational motion. OBJECTIVE: To reveal whether women with chronic whiplash-associated disorders, Grades I-II, demonstrate evidence of abnormal segmental motions in the cervical spine. SUMMARY OF BACKGROUND DATA: It is hypothesized that unphysiological spinal motion experienced during an automobile accident may result in a persistent disturbance of segmental motion. It is not known whether patients with chronic whiplash associated disorders differ from patients with chronic insidious onset neck pain with respect to segmental mobility. METHODS: Lateral radiographic views were taken in assisted maximal flexion and extension. A new measurement protocol determined rotational and translational motions of segments C3-C4 and C5-C6 with high precision. Segmental motion was compared with normal data as well as among groups. RESULTS: In the whiplash-associated disorders group, the C3-C4 and C4-C5 segments showed significantly increased rotational motions. Translational motions within each segment revealed a significant deviation from normal at the C3-C4 segment in the whiplash-associated disorders and insidious onset neck pain groups and at the C5-C6 segment in the whiplash-associated disorders group. Significantly more women in the whiplash-associated disorders group (35.3%) had abnormal increased segmental motions compared to the insidious onset neck pain group (8.6%) when both the rotational and the translational parameters were analyzed. When the translational parameter was analyzed separately, no significant difference was found between groups, or 17.6% (whiplash-associated disorders group) and 8.6% (insidious onset neck pain group), respectively. CONCLUSION: Hypermobility in the lower cervical spine segments in 12 out of 34 patients with chronic whiplash-associated disorders in this study point to injury caused by the accident. This subgroup, identified by the new radiographic protocol, might need a specific therapeutic intervention. PMID- 14520036 TI - Anterior single rod instrumentation for thoracolumbar adolescent idiopathic scoliosis with and without the use of structural interbody support. AB - STUDY DESIGN: A radiographic and clinical outcomes analysis of 41 patients treated for thoracolumbar adolescent idiopathic scoliosis utilizing a single anterior rigid rod construct. OBJECTIVES: To evaluate the necessity of structural interbody support to improve primary curve correction and preserve or augment lordosis when used in conjunction with a single anterior rigid rod construct, to identify parameters that predict horizontalization of the lowest instrumented vertebra, adjacent disc angulation, and distal uninstrumented vertebrae, and to assess patient satisfaction following surgery. BACKGROUND DATA: Instrumentation induced kyphosis has been a concern with nonrigid anterior systems used in the past for the treatment of scoliosis. Interbody structural support has been recommended to maintain appropriate sagittal profile when anterior systems are utilized. It has also been suggested that the use of structural interbody support creates a fulcrum to increase curve correction when compression is applied to the convexity of the deformity. However, the necessity of interbody structural support when used in conjunction with a rigid anterior system has not been previously evaluated in patients with adolescent idiopathic scoliosis. MATERIALS AND METHODS: Forty-one patients mean age 15.9 years (range 12.1-18.6 years) with thoracolumbar adolescent idiopathic scoliosis underwent anterior spinal fusion using a single 6.0 to 6.5 mm solid rod construct between June 1995 and August 1999 performed by the senior author (T.G.L.). Four additional patients with thoracolumbar curves with similar anterior instrumentation over the same time period were lost to follow-up or had incomplete records and were not included in the study. Structural interbody support was used in 21 patients and packed morselized autograft alone was used in 20 patients. The patients in the group with packed morselized bone alone generally underwent surgery earlier in the series before the author began using structural interbody support on a regular basis. Each patient had a minimum follow-up of 3 years. Preoperative, initial, and most recent (>3 years) follow-up radiographs were reviewed to determine in each group Cobb angle measurements, flexibility of primary, secondary, and fractional curves, apical and end vertebral translation, lowest instrumented vertebral and caudal disc angulation, global coronal and sagittal balance, and sagittal Cobb measurements in both instrumented levels as well as lumbar lordosis (T12-S1). In addition, the SRS outcomes instrument was completed by 38 of 41 patients. RESULTS: The mean preoperative primary curve in patients with structural support was 47 degrees (Group II) and 45 degrees in patients without structural support (Group I). Mean curve correction was to 13 degrees in Groups I and II. One patient in Group II became slightly more unbalanced at final follow up; otherwise all were improved after surgery. Sagittal measurements over instrumented segments as well as total lumbar lordosis (T12-S1) was maintained between preoperative and final postoperative values in both groups. Similarly, in both groups, when horizontalization of the distal end instrumented vertebra was achieved on the preoperative reverse side-bending radiograph, more normal relationships were achieved between instrumented and distal noninstrumented segments (adjacent disc angulation and fractional lumbar curve) at final follow up (P 30 degrees. RESULTS: Patients with tight hamstrings have a significantly greater risk of postoperative imbalance (P = 0.05), and these patients can only compensate for this risk by reducing their lumbar lordosis (P = 0.0227). Furthermore, the limitations in the lumbar and pelvic range of motion are predicted by tight hamstrings (P 5 degrees of correction, with a mean improvement in sagittal alignment of this population of 14.2 degrees. Patient surveys revealed significant pain reduction within the first week after surgery and improved activity levels for a majority of patients. CONCLUSIONS: Kyphoplasty improves physical function, reduces pain, and may correct kyphotic deformity associated with vertebral compression fractures. PMID- 14520044 TI - Cluster analysis classification of SF-36 profiles for patients with spinal pain. AB - STUDY DESIGN: A k-means cluster analysis of patients with spinal and radicular pain based on the SF-36 Health Survey scales. OBJECTIVE: The aim was to determine whether spine patients fall into clusters according to self-reported health status as measured by the SF-36 and to determine if clustering is similar across four common diagnostic categories: herniated disc, spinal stenosis, spondylosis, and chronic pain syndrome. SUMMARY OF BACKGROUND DATA: Cognitive-behavioral classifications of chronic pain patients have previously identified three patient groups described as Dysfunctional, Interpersonally Distressed, and Minimizers/Adaptive Copers. The purpose of these classifications is to facilitate and direct treatment based not only on biomedical diagnosis but also on emotional, social, and behavioral diagnoses. This type of analysis has not been done on the quality-of-life scores of patients with specific spinal diagnoses. METHODS: Health status data were reviewed from the initial visits of 15,748 spine patients in the National Spine Network database. Based on the eight scales of the SF-36, k-means cluster analysis divided the National Spine Network population into distinct clusters of similar patients. Clustering was performed separately for each clinical diagnosis group. RESULTS: In all four diagnostic categories, cluster analysis classified patients into three groups. Group 1 had fairly high (relative to the entire sample) scores on all scales and was labeled "Highly Functional." Group 2 had low measures on physical variables but comparatively high scores on the mental scales. These were labeled "Emotional Adapters." Group 3 had low scores on all scales. These patients were labeled "Dysfunctional." Although patients in each diagnostic category fell into one of the three groups, the proportion of patients within each group was quite different among chronic pain patients as compared to the other three diagnostic groups. For example, 29% of herniated disc patients were in the Highly Functional group, whereas only 14% of patients in chronic pain were categorized as Highly Functional. Thirty-three percent of spondylosis patients were classified as Dysfunctional compared with 51% of chronic pain patients. CONCLUSIONS: Patients with spinal pain fall into three groups according to their profile of scores on the SF-36 Health Survey. It is proposed that such empirical groupings can guide decision-making in selecting the most appropriate therapies. PMID- 14520045 TI - Recurrence of low back pain: definition-sensitivity analysis using administrative data. AB - STUDY DESIGN: Retrospective analysis of administrative claims data in a single workers' compensation jurisdiction. OBJECTIVES: To evaluate the effects of alternative definitions and follow-up parameters on rates of low back pain recurrence based on detailed administrative data. SUMMARY OF BACKGROUND DATA: Previous studies reported low back pain recurrence rates ranging from 14% to 45%, without consistency in definitions of recurrence or specifications of follow-up. METHODS: Patients with new claims for low back pain reported in New Hampshire to a large workers' compensation provider in 1996 and 1997 were selected (N = 2944). Definitions of recurrence included: new workers' compensation claim, new episode of care, and new episode of lost work time (work disability). For the latter two definitions, various minimum between-episode gaps were applied and related to recurrence rates. Two follow-up structures (constant length of follow-up post end of the first episode and fixed-period length of follow-up since the onset of low back pain) were examined for sensitivity of recognizing low back pain recurrence, with a maximum of 3-year follow-up. RESULTS: Recurrence rate using a claims-based definition was 7.9% and 7.1% for the entire cohort and the subset with work disability days, respectively, for the 3 years of follow-up. Care-based recurrence rates ranged between 12% and 49%, whereas disability-based recurrence rates ranged between 6% and 17% over the 3 years, inversely related to the length of the minimum between-episode gap (R = -0.86 for disability and care, P < 0.001). Two-year follow-up was sufficient to identify 85% to 100% of recurrences regardless of the follow-up structure. CONCLUSIONS: Recurrence rates are highly sensitive to variations in definitions. Consistency of definitions and application across studies is required to enable valid comparisons. PMID- 14520046 TI - Familial disability patterns in individuals with chronic work-related spine injury/illness. AB - STUDY DESIGN: A case control study of familial disability patterns and other potential risk factors identified in a group of individuals with work-related spine injury/illness. OBJECTIVES: The objective of the present study is to provide information on the relationship between a disabled family member and work related spine injuries or illnesses leading to disability among another family member. SUMMARY OF BACKGROUND DATA: Extensive studies have been conducted to identify the medical and nonmedical factors associated with prolonged disability after work-related spine injury/illness. Many of the relevant nonmedical factors are psychosocial and are dependent to some degree on family of origin environment and experience. Although similar recurring pain behaviors have been described in certain families, no recent study has been conducted of coexisting family disabilities in individuals with work-related spine injury/illness. METHODS: A case-control investigation examining nine potential risk factors, including familial disability patterns, for the development of disability following work related spine injury/illness was conducted. The case group (n = 88) consisted of individuals disabled from any and all gainful employment, whereas the controlled group (n = 40) continued working in spite of their spine complaints. RESULTS: In this study, gender, number of spinal areas involved, current injury considerations/severity, and having a primary family member or spouse similarly disabled were found to be significant predictors of disability following work related spine injury/illness. However, further analysis revealed that having an associated family disability was the greatest predictor of disability in the study group. CONCLUSIONS: These data suggest that the presence of a primary family member or spouse with a work-related spine disability may be a risk factor for the development of disability in individuals sustaining a work-related spine injury. PMID- 14520047 TI - Prevention of cerebrospinal fistulae and reduction of epidural scar with new surgical hemostat device in a porcine laminectomy model. AB - STUDY DESIGN: In a porcine laminectomy model, a standard dural/arachnoid incision was made and tested for cerebrospinal fluid leak after material application. Sites were graded for scar formation and healing response at 3 weeks. OBJECTIVE: This study compares effectiveness of CoStasis, Tissucol, and suture for prevention of cerebrospinal fluid leaks and epidural scar formation after spinal dural incisions. SUMMARY OF BACKGROUND DATA: Cerebrospinal fluid leaks following cranial and spinal surgery are potentially serious complications. Epidural scar formation is exacerbated by improper control of hemostasis. A hemostatic agent with dural sealant properties may be advantageous. METHODS: Total laminectomy was performed at three levels in seven pigs. At each level, a uniform 1.5 cm incision was made in the dura and arachnoid. A single suture was placed to approximate the edges and sites were treated with one of three methods: CoStasis, Tissucol, or no treatment. At sacrifice, 3 weeks later, epidural scar was graded, pressure testing of some sites was done, and tissue for histologic sections was harvested. RESULTS: CoStasis and Tissucol produced immediate dural sealing when the valsalva maneuver was applied. One suture-only site leaked. At sacrifice, all sites were sealed. CoStasis and Tissucol had less scar formation than control sites. Pressure testing results were similar at CoStasis and Tissucol sites. CONCLUSION: CoStasis and Tissucol have comparable effectiveness in sealing CSF leaks immediately and at 3 weeks after complete laminectomy. CoStasis demonstrated comparable performance to Tissucol with less epidural scar formation than primary suture alone. PMID- 14520048 TI - Analysis of porous ingrowth in intervertebral disc prostheses: a nonhuman primate model. PMID- 14520049 TI - Re: Walsh TL, Hanscom B, Lurie JD, et al. Is a condition-specific instrument for patients with low back pain/leg symptoms really necessary? The responsiveness of the Oswestry Disability Index, MODEMS, and the SF-36. Spine 2003;28:607-15. PMID- 14520051 TI - Physical conditioning programs for workers with back and neck pain: a cochrane systematic review. AB - OBJECTIVE: To determine the effect on time lost from work of physical conditioning programs for workers with back and neck pain. DATA SOURCES Randomized trials were located by searching MEDLINE, EMBASE, CINAHL, PsycINFO, the Cochrane Controlled Trial Register, and PEDro. REVIEW METHODS: Two reviewers independently extracted data and assessed trial quality. Where data could be pooled, meta-analysis was performed. Based on cost considerations, we nominated a mean saving of 10 sick days per year or a number needed to treat to return 1 person to work of 10 as the smallest treatment effects that would be clinically worthwhile. RESULTS: Nineteen trials in 21 publications yielded 23 contrasts relevant to this review. These trials provide evidence that physical conditioning programs that included a cognitive-behavioral approach could produce a clinically worthwhile reduction in the number of sick days taken at 12 months (average of 45 days; 95% confidence interval 3-88) when compared to general practitioner care or advice for workers with chronic back pain. There was little evidence of an effect on time lost from work of specific exercise programs that did not include a cognitive-behavioral component. CONCLUSION: Physical conditioning programs that incorporate a cognitive-behavioral approach reduce the number of sick days for workers with chronic back pain when compared to usual care. PMID- 14520050 TI - A ligamentum flavum-preserving approach to the lumbar spinal canal. AB - STUDY DESIGN: This is a retrospective study of a consecutive series of patients undergoing spinal decompression or disc enucleation with preservation of the ligamentum flavum. MacNab criteria and Prolo scoring was used to assess them pre- and after surgery. OBJECTIVES: The aim of the study was to assess the incidence of perioperative complications and long-term clinical outcome of the procedure. SUMMARY OF THE BACKGROUND DATA: One previous study has shown this technique to be effective in reducing the rate of perioperative complications and recurrences in patients undergoing microscopic disc enucleation. However, this study does not comment on the long-term clinical results or the use of the technique in a wider range of spinal procedures. METHODS: Between 1991 and 1999, we operated on 214 patients for peripheral lumbar radiculopathy. MacNab criteria and Prolo scoring was used for their pre- and postoperative assessment. Postoperative assessment was with the help of a postal questionnaire. In addition, clinical notes were studied to look for any further procedures or operative complications. RESULTS: All 214 patients returned the postoperative questionnaires. There were 104 males and 110 females. The mean follow-up was 5.97 years. Thirty-eight patients had an associated fusion or Graf ligament stabilization. Two hundred ninety-six levels were operated in all. There were four instances of dural tear and one of nerve root injury. Five patients underwent revision. There was a statistically significant improvement in the MacNab criteria (P < 0.001) and Prolo score (P < 0.001) after surgery. CONCLUSIONS: We recommend a ligamentum flavum preserving approach because it decreases the rate of perioperative complications while giving comparable long-term results. PMID- 14520052 TI - Synovial chondromatosis presenting with cervical radiculopathy: a case report. AB - STUDY DESIGN: A case report is presented. OBJECTIVES: To report a case of synovial chondromatosis of a cervical facet joint and describe the appearance with magnetic resonance imaging and computed tomography myelography. SUMMARY OF BACKGROUND DATA: Synovial chondromatosis is an uncommon disorder characterized by the presence of multiple cartilaginous or osteocartilaginous nodules in the synovium of a joint space. Synovial chondromatosis in the cervical facet joint is rare. METHOD: A 52-year-old woman experienced the sudden onset of severe pain in the dorsal shoulder girdle and in the ulnar side of her right arm and forearm. This refractory pain only responded to an epidural nerve root block. Neurologic examination showed right nerve root signs that ranged from the C7 to Th1 segments of the spinal cord. Radiologic and electrophysiological examinations were carried out. RESULT: A mass was found in the right facet joint between C7 and Th1 with magnetic resonance imaging and computed tomography myelography. These investigations clearly indicated the location, size, and extent of the lesion accompanying the irregularity of the joint and osteolytic change. Somatosensory evoked potentials with right ulnar nerve stimulation indicated a significant conduction block in the lower right cervical nerve roots. After surgical removal of this lesion, the neurologic symptoms markedly improved. The histopathology diagnosed synovial chondromatosis. CONCLUSION: Synovial chondromatosis should be included in the differential diagnosis of radiculopathies of unknown etiology. PMID- 14520053 TI - Radiculopathy due to ossification of the yellow ligament at the lower lumbar spine. AB - STUDY DESIGN: A case report. OBJECTIVES: To report a rare case of a 27-year-old female with ossification of yellow ligament at the lower lumbar spine presenting radiculopathy with a drop foot. SUMMARY OF BACKGROUND DATA: The majority of cases of ossification of yellow ligament occur at the lower third of the thoracic or the thoracolumbar spine. There are only a few reports of ossification of yellow ligament in the lumbar spine and radiculopathy due to ossification of yellow ligament at L4-L5 and L5-S1 levels is very uncommon. METHODS: A 27-year-old female with a prior fracture of posterior ring apophysis of L5 presented with leg pain and a drop foot. Magnetic resonance imaging demonstrated stenosis with compression of the cauda equina at the L4-L5 and L5-S1 levels. RESULTS: Decompressive laminectomy of L5 and removal of the ossified yellow ligaments were performed. Histologic examination of en bloc specimen of ossification of yellow ligament revealed degenerative changes of the elastic fibers in the yellow ligament with adjacent chondrosis and ossification. The patient's severe leg pain disappeared completely, although the extent of the drop foot had not fully recovered at the final follow-up examination. CONCLUSIONS: The mechanism of ossification of yellow ligament in the present case was unclear. The patient did not have any previous generalized disorders besides the history of a ring apophysial fracture or any family history of treatment for ossification of the posterior longitudinal ligament or ossification of yellow ligament. Therefore, localized mechanical stress might have influenced the development of ossification of yellow ligament at lower lumbar spine. PMID- 14520054 TI - The extraforaminal juxtafacet cyst as a rare cause of L5 radiculopathy: a case report. AB - STUDY DESIGN: This is a report of a case. OBJECTIVE: To document the clinical, radiographic, and histologic characteristics of a lumbar extraforaminal juxtafacet cyst. SUMMARY OF BACKGROUND DATA: Spinal juxtafacet cysts develop most frequently at the dorsal aspect of the zygapophysial joint, sometimes in the posterolateral area of the canal. In one case, they have been described in the foraminal and extraforaminal region. METHODS: Description of the case report. RESULT: The authors report one case of a strictly extraforaminal juxtafacet cyst responsible for L5 sciatica. CONCLUSIONS: Juxtafacet cysts of the spine represent an infrequent cause of sciatica, usually when they grow in the canal, or more exceptionally when they occupy the foraminal or extraforaminal areas. PMID- 14520055 TI - Primitive neuroectodermal tumor and extraskeletal Ewing sarcoma arising primarily around the spinal column: report of four cases and a review of the literature. AB - STUDY DESIGN: Report of four cases and a review of the literature. OBJECTIVES: To study the clinical features and prognosis of primitive neuroectodermal tumor or extraskeletal Ewing sarcoma arising around the spinal column. SUMMARY OF BACKGROUND DATA: Primitive neuroectodermal tumor or extraskeletal Ewing sarcoma that originates around the spinal column is very rare, and its prognosis is very poor. METHODS: Four patients were diagnosed and underwent treatment. RESULTS: Although all the patients received high-dose chemotherapy with or without radiotherapy after surgery, three patients died of the disease. Only one patient who received en bloc resection of the tumor combined with multiagent chemotherapy followed by high-dose chemotherapy with peripheral blood stem cell transplantation remains alive and continues to be disease free. CONCLUSION: The prognosis of the patients with primitive neuroectodermal tumor or extraskeletal Ewing sarcoma around the spinal column is very poor. Multiagent chemotherapy combined with en bloc resection and radiation therapy is the preferred treatment for patients with primitive neuroectodermal tumor or extraskeletal Ewing sarcoma around the spinal column. PMID- 14520056 TI - Causes of death among Finnish women with cosmetic breast implants, 1971-2001. AB - Mortality patterns among women with cosmetic breast implants have not been well characterized. We examined cause-specific mortality among women who underwent cosmetic breast implantation at major public hospitals and private clinics in Finland from 1970 through 2000. Causes of death through 2001 were identified through the national mortality register. Expected numbers of deaths were calculated on the basis of mortality rates in the general female population, and standardized mortality ratios (SMR), i.e. ratios of observed to expected deaths, and 95% confidence intervals (CI) were calculated. Among the 2166 women with cosmetic breast implants, we observed 31 deaths versus 32.1 expected. Overall disease mortality was below expectation, mainly due to low mortality from diseases of the circulatory system (SMR, 0.28; 95% CI, 0.03-1.02). Mortality from cancer was close to expectation. There was a statistically significant excess of suicide, based on 10 deaths, which was most pronounced during the first 5 years of follow-up (SMR, 4.26; 95% CI, 1.56-9.26). In conclusion, although based on small numbers, women with cosmetic breast implants did not experience higher mortality overall than women in the general population. The high suicide risk supports other studies and warrants further detailed investigation. PMID- 14520058 TI - An application of the LeJour vertical mammaplasty pattern for skin-sparing mastectomy: a preliminary report. AB - An application of the LeJour vertical mammaplasty skin pattern for skin-sparing mastectomy is presented. The approach provides adequate access for the mastectomy, axillary dissection, and immediate breast reconstruction. The technique is ideal for patients with large or ptotic breasts undergoing a simultaneous contralateral breast reduction or mastopexy. It is particularly suitable for autogenous tissue reconstruction. Its use in mastectomies for cancer and prophylactic subcutaneous mastectomies is described. PMID- 14520060 TI - Is it safe to combine abdominoplasty with other dermolipectomy procedures to correct skin excess after weight loss? AB - The development of sophisticated bariatric surgery techniques has led to an increasing number of patients demanding a complexity of body contouring procedures that can be achieved either with a step-by-step approach or by combining dermolipectomy in various regions. The aim of this study was to test the hypothesis that abdominoplasty-related morbidity and outcome may be worsened by combining abdominoplasty with other dermolipectomies and to determine whether any predictive risk factor could be identified. Seventy-three consecutive patients undergoing abdominoplasty with (n = 26) and without (n = 47) additional dermolipectomy on the upper arms, inner thighs, breasts, and buttocks were included in this study. Additional dermolipectomies did not increase abdominoplasty-related morbidity but revealed better long-term results (p = 0.10). Statistically significant risk factors were male sex, age (>41 years), overweight (BMI > 30), prolonged operations (>3 hours), and extensive blood loss (1,000 mL). Most importantly, better outcome was obtained in patients whose weight reduction was greater (p = 0.04). The authors' results suggest that combined dermolipectomy procedures can be advocated in patients who have reached their intended level of weight reduction and if prolonged operative time and excessive blood loss are avoided. PMID- 14520061 TI - Frequency and distribution of talc contamination in patients with silicone gel filled breast implants. AB - Talc was found to contaminate peri-implant scars in patients with silicone gel filled breast prostheses resulting from the physical interaction between implant and surgical glove. This study reports the frequency and distribution of the problem and its relationship to changes in glove manufacture. Peri-implant scar tissue, obtained from a population with breast implants, was evaluated for talc. Patients were considered positive if more than two talc particles were seen intracellularly in more than two microscopic fields. The largest Texas glove manufacturer stopped using talc in 1983. Patients were grouped to demonstrate the difference in talc exposure in Texas after this manufacturing change. Of those receiving implants before 1984, 136 of 140 were positive for talc. Of those receiving implants after 1983, 24 of 54 were positive (p < 0.000; RR = 42.5; CI: 13.7-131.6). Widespread prevalence of contamination was evidenced by findings of talc in patients from various facilities, different surgeons, and multiple sites within Texas. PMID- 14520062 TI - Improving recipient vessel exposure during microvascular breast reconstruction. AB - Microvascular tissue transfer has become the gold standard for breast reconstruction. The primary drawback to these procedures is the technical expertise required for microsurgical anastomosis. This problem is compounded by the difficulties in the exposure of recipient vessels deep within the axilla. Previous techniques used for exposure of these vessels are difficult to setup, provide less than optimal exposure, and have been associated with brachial plexus injuries. The authors retrospectively review their experience using the pediatric OMNI retractor for exposure of recipient vessels during microvascular breast reconstruction. Patient demographics, flap choice, recipient vessels, the incidence of neuropraxia/brachial plexopathy, and microvascular complications were analyzed. Patients in whom more traditional methods of vessel exposure were used (ie, Gelpi retractors, arm positioning, fish hooks; 517 reconstructions in 392 patients) were compared with patients in whom vessel exposure was performed using the pediatric OMNI retractor (699 reconstructions in 571 patients). No differences were noted in comorbid conditions or the incidence of microvascular complications. However, the use of the pediatric OMNI was associated with a significant reduction in operative time in unilateral reconstructions (6:23 +/- 0.05 h vs 7:48 +/- 0.05 h; P <0.01) and decreased incidence of brachial plexus injury (0.17% vs 3.3%; P <0.01). The authors think the decreased neuropraxia rate is the result of better exposure afforded by the pediatric OMNI retractor, which improves exposure and eliminates the need for excessive arm abduction or awkward positioning during the dissection and anastomosis of axillary recipient vessels. PMID- 14520063 TI - The flashlamp-pumped pulsed dye laser (585 nm) treatment of hypertrophic scars in Asians. AB - Hypertrophic scars are common and cause functional and psychologic morbidity. Various treatment modalities have been advocated in the past, but the flashlamp pumped pulsed-dye laser-585 nm (PDL) has been shown to be effective in the treatment of a variety of traumatic and surgical scars. This study was performed to determine the effectiveness of the PDL treatment of hypertrophic scars in Asians and to elucidate possible side effects and complications. Thirteen Asian patients with 19 hypertrophic scars that had no response to conservative therapies, such as pressure therapy, intralesional corticosteroid injection, or silastic gel sheeting for more than 6 months, were treated with the PDL. Three months after the last laser treatment, improvement in the appearance, reduction in scar height, and erythema as a result of the PDL treatment were evaluated and compared with baseline values. Compared with the baseline appearance, hypertrophic scars treated with the PDL showed a satisfactory clinical improvement of 84% (16 of 19), a significant percentage of scar flattening of 58.9 +/- 22.1%, and of erythema elimination of 75.7 +/- 19.9% at month 3 after the last laser treatment. The PDL treatment of hypertrophic scars can effectively improve scar pliability and texture and decrease erythema and associated symptoms. In addition, multiple treatment sessions are suggested for achieving greater response, but lower fluences are recommended for prevention of side effects in Asians with a higher melanin content of the skin. PMID- 14520064 TI - The soft-tissue shaving procedure for deformity management of chronic tophaceous gout. AB - Gout is a condition characterized by the deposition of monosodium urate crystals in the joints or soft tissue. A gouty tophus occasionally mimics an infectious or neoplastic process. However, the conventional enucleating procedure might cause complications. In severe cases, skin necrosis and tendon or joint exposure can occur after debridements. In this series, the soft-tissue shaver is used for deformity management of the chronic tophaceous patients and the results are encouraging. From November 2000 to August 2002, 17 patients with chronic tophaceous gout were treated by the shaver technique. Suction and irrigation were performed simultaneously while the shaver was operating, and chalky deposits of sodium urate could be removed efficiently. Skin necrosis was minimized by means of proper incision planning. Also, bedside debridements and wound wet dressing were helpful for improving the outcomes. The families and patients were satisfied with the results. In conclusion, severe chronic tophaceous gout can be a surgical challenge. The soft-tissue shaving technique can be useful for cosmetic debulking of large tophi in patients with advanced chronic tophaceous gouty arthritis. PMID- 14520065 TI - Reconstruction of combined defects of the Achilles tendon and the overlying soft tissue with a fascia lata graft and a free fasciocutaneous lateral arm flap. AB - A new approach to reconstruction of the Achilles tendon and overlying soft tissue is presented. A fascia lata graft is used to reconstruct the tendon and is enwrapped by the fascia that is included in a fasciocutaneous lateral arm flap. Five patients were treated with this technique; three of them after surgical Achilles tendon repair, rerupture, and consecutive infection, one after a full thickness burn with loss of the tendon and one with a history of ochronosis and necrosis of the whole tendon and overlying soft tissue. There were no anastomotic complications and all flaps healed primarily. Functional evaluation with the Cybex II dynamometer was done at least 49 months after reconstruction. A good functional and cosmetic result was obtained in all patients and donor site morbidity was acceptable. These results are well within the results of other surgical treatment options reported in the literature. PMID- 14520066 TI - Regenerative surgery for sacral pressure ulcers using collagen matrix substitute dermis (artificial dermis). AB - Collagen matrix substitute dermis, or artificial dermis, has recently been developed to induce angiogenesis and fibroplasia in deep, poorly vascularized tissue defects, and its use is desirable as a means of achieving effective treatment with less invasion. However, it is difficult to apply collagen matrix to pressure ulcers, because they are usually accompanied by infection with discharge of excessive amounts of exudate or pus and generally exposed to external forces that prevent graft fixation. To overcome these difficulties, the authors demonstrated a novel procedure to exclude unfavorable exudate and external forces. The procedure resulted in successful fixation of the artificial dermis and induction of regeneration in poorly vascularized defects in every case. The histologic findings provided supporting evidence that collagen matrix acts as a scaffold for reconstruction of nearly normal vascular structures coursing perpendicularly in the upper layer of normal dermis. PMID- 14520067 TI - Three-dimensional nasal morphology in cleft lip and palate operated adult patients. AB - Nasal morphology was assessed three-dimensionally in 18 cleft lip and palate operated patients aged 19 to 27 years, and in 161 sex- and age-matched control subjects. Comparisons were performed by Student t and Watson-Williams' tests. No differences were found in nasal volume or surface. Nasal width, alar base width, and inferior width of the nostrils were significantly larger in male patients than in the reference men. The nasal bridge was shorter. Similar differences were found in the female patients. In addition, the right nostril was larger and the nasal height shorter. In men, the nasolabial and the nasal tip angles were smaller in the patients than in the reference subjects, whereas the facial convexity angle was larger in the patients. In conclusion, the nose of adult operated cleft lip and palate patients differed from that of normal control subjects. Surgical corrections of the cleft lip and palate failed to provide a completely normal appearance. The methods might be used to indicate where additional procedures might be performed to approximate a reference population. PMID- 14520068 TI - Venous "supercharging" augments survival of the delayed rat TRAM flap. AB - Adequate delay of a pedicled transverse rectus abdominis musculocutaneous (TRAM) flap might not necessarily require interruption of the venous system. The retained ipsilateral deep vein of the dominant pedicle could then be used as a secondary outflow source for potential salvage of a congested flap. A venous "supercharged" rat TRAM flap model has been designed to evaluate the efficacy of this maneuver. Seventy-two female Sprague-Dawley rats (CD) were equally divided into two major groups, differing only in whether a delay by division of the dominant cranial epigastric artery had first been performed. An inferior-based TRAM flap (nondominant) was raised for each rat, with three subsets of 12 rats in each group, ie, with the cranial epigastric vein subsequently divided (group control), retained (supercharged), or retained but the inferior pedicle divided (venous flap). Both supercharged subsets had significantly augmented flap survival when compared with flaps in their group raised without the cranial epigastric vein, whether a delay maneuver had (96 +/- 6% vs 89 +/- 7%; P = 0.012) or not (80 +/- 8% vs 65 +/- 21%; P = 0.034) been performed. Flaps with only a cranial epigastric vein pedicle totally necrosed, implying that the observed enhancement in flap viability was not the result of transformation into a venous flap, but perhaps as a crossover flap where an adjacent venosome was captured. Venous supercharging can be accomplished by inclusion of the ipsilateral dominant deep vein, and should be a consideration in the clinical planning of delay maneuvers and for treatment of the compromised TRAM flap. PMID- 14520069 TI - A vertical midline scar is a 'high-risk' factor for maximum survival of the rat TRAM flap. AB - The presence of any abdominal scar, in addition to obesity, a smoking history, and prior irradiation are considered the major known "risk factors" for predictable success or failure of the lower transverse rectus abdominis musculocutaneous (TRAM) flap. For many, a vertical midline scar has even been considered to be a relative contraindication. The possibility that the scar instead could effect some form of delay or by neovascularization permit reperfusion across the midline might negate this concern. The validity of this hypothesis was tested in 40 Sprague-Dawley (CD) rats using our standard rat TRAM flap model. Every rat initially had a vertical skin incision made from xiphoid to pubis. At a second stage, either immediately or after a delay of 1 week, 2 weeks, or 6 months, a superior-pedicled (dominant) or inferior-pedicled (nondominant) TRAM flap was raised, with five rats in each subgroup. For the inferior-pedicled group, the percentage of ipsilateral (muscle-pedicle half) flap survival approached 75% and had a trend toward greater survival with each increase in the time of delay, but any difference was not statistically significant (F= 0.653, P = 0.538). In the superior-pedicled group, the ipsilateral half of the flap always survived completely. In both groups, the contralateral or opposite side always underwent complete necrosis regardless of pedicle orientation or time constraints. The midline scar did not enhance even unilateral TRAM flap survival when compared with historic controls, and long-term transmidline reperfusion across the scar did not seem to occur. These findings corroborate the clinical observation that only a unilateral TRAM flap would be reliable in the presence of a vertical midline abdominal scar. PMID- 14520070 TI - Accelerated diabetic wound healing using cultured dermal fibroblasts retrovirally transduced with the platelet-derived growth factor B gene. AB - The treatment of diabetic wounds is a considerable clinical challenge. In this study, mouse dermal fibroblasts retrovirally transduced with the human platelet derived growth factor B (PDGF-B) gene were used to treat diabetic mouse wounds. The PDGF-B gene was obtained from human umbilical vein endothelial cells, cloned into retroviral vectors, and introduced into diabetic mouse C57B1/ks-db/db dermal fibroblasts. In vitro results demonstrated production of PDGF-B protein by these transduced cells at steady-state levels of 1000 ng PDGF-B/10(6) cells/24 hours, and expression of PDGF-B mRNA. These cells were seeded onto polyglycolic acid scaffold matrices and used to treat diabetic mouse 20-mm x 20-mm full-thickness excisional dorsal skin wounds. Measurement of the residual epithelial gap at 21 days showed significantly accelerated healing (P < 0.05) of wounds treated with PDGF-transduced cells (epithelial gap 10.46 +/- 1.20 mm) compared with untreated wounds (14.66 +/- 0.591 mm), wounds treated with polyglycolic acid alone (14.80 +/- 0.575 mm), or wounds treated with negative control LNCX-transduced cells (13.76 +/- 0.831 mm). Immunohistochemical staining showed intense staining for PDGF in wounds treated with PDGF-B-transduced cells. This study demonstrates the promising potential for gene therapy in diabetic wound healing. PMID- 14520071 TI - Silicone gel breast implant rupture presenting as a fluctuant back mass after latissimus dorsi breast reconstruction. AB - The authors describe a patient who presented with acute onset of a lower back fluctuant mass 12 years after breast reconstruction with a latissimus dorsi musculocutaneous flap and silicone gel implant. Aspiration and subsequent surgical exploration revealed this mass to be free-flowing silicone gel within a cavity that was confluent with the breast implant capsule through an axillary tunnel. Excision of the back cavity, explantation with subtotal capsulectomy, and implant replacement resolved the problem. Although distant migration of extracapsular silicone gel from the breast to the axilla, arm, abdomen, and groin has been described, the authors think this represents the first reported case of distant migration of silicone gel to the lower back. PMID- 14520072 TI - Hatchet-shaped tensor fascia lata musculocutaneous flap for the coverage of trochanteric pressure sores: a new modification. AB - The tensor fascia lata flap is one of the appropriate choices for the coverage of trochanteric pressure sores. The authors designed a new, hatched-shaped tensor fascia lata musculocutaneous flap with distal Z-plasty closure and applied it to four trochanteric defects in 4 patients. Satisfactory results were obtained in all patients. The hatchet-shaped tensor fascia lata musculocutaneous flap is very safe, reliable, and practical. Designing the flap in a hatchet shape allows one to use the proximal and well-vascularized portion of the flap in the trochanteric pressure sore area. Another important advantage is the possibility of reuse resulting from recurrence. Prevention of a "dog-ear" deformity at the recipient site provides a smooth contour on the lateral aspect of the thigh. Another advantage is the tension-free Z-plasty closure of the donor site without need of grafting. PMID- 14520073 TI - Two-stage reconstruction of infected deep median sternotomy wound with an intercostal pedicled oblique rectus abdominis musculocutaneous flap after coronary artery bypass grafting. AB - Median sternotomy wounds infected after coronary artery bypass grafting with bilateral internal mammary arteries are generally poor in condition, cannot be debrided adequately, and are limited in flap selection for reconstruction. The authors treated 2 patients with two-stage reconstruction using a modified superior-based rectus abdominis musculocutaneous flap. First, simple debridement was performed with the goal of preserving the internal mammary artery grafts. Then, delayed reconstruction with the oblique rectus abdominis musculocutaneous flap supplied by the superior epigastric and seventh intercostal vessels was performed. Despite ligature of the bilateral internal mammary arteries at their full length, the large oblique skin paddle designed along the angiosome as far as the midaxillary line survived almost completely in both patients, resulting in cessation of pus discharge. The circulation to the superior epigastric and intercostal vessels might be reinforced because of the delay phenomenon. Two stage reconstruction with a superior pedicled oblique rectus abdominis musculocutaneous flap presents a successful resolution of infected median sternotomy wounds after coronary artery bypass grafting with sacrifice of bilateral internal mammary arteries. PMID- 14520075 TI - Inalienably right. PMID- 14520074 TI - A new method for vaginal reconstruction using a pedicled jejunal flap. AB - A technique for vaginal reconstruction is described that uses a pedicled jejunal flap. A neovagina was constructed using a segment of jejunum based on the fourth branch of the superior mesenteric artery. This provided an excellent result with adequate length, without the need for a lubricator or stent. Hypersecretion has not been observed. PMID- 14520076 TI - Re: A simple method for cranial bone chip harvesting. PMID- 14520078 TI - Degas is dead...listen. PMID- 14520077 TI - Extra-articular ankylosis of the mandible caused by possible migration of bone grafts. PMID- 14520079 TI - Cervical burners in the athlete. PMID- 14520080 TI - 13-year-old with back and shoulder pain. PMID- 14520081 TI - Vaginal discharge in the prepubertal child. PMID- 14520082 TI - Pediatric pharyngitis. PMID- 14520083 TI - Evaluation and management of an infantile esotropia. PMID- 14520084 TI - This treat is not so sweet: exploratory sulfonylurea ingestion by a toddler. PMID- 14520087 TI - Presidential address to the Triological Society. PMID- 14520085 TI - The child with aseptic meningitis. PMID- 14520088 TI - Long-term effect of adenotonsillectomy on quality of life in pediatric patients. AB - HYPOTHESIS: Adenotonsillectomy improves general and disease-specific quality of life for properly selected patients suffering for upper airway obstruction secondary to adenotonsillar hypertrophy (UAO) and obstructive sleep apnea (OSA). STUDY DESIGN: Prospective, nonrandomized questionnaire. SPECIFIC AIMS: To evaluate quality of life in patients with UAO and OSA using general quality of life instruments as well as disease-specific instruments. The results will be compared both pre- and postoperatively. METHODS: Fifty-five patients aged 2 to 16 with the clinical diagnosis of UAO or OSA were recruited. The caregivers completed Children's Health Questionnaire Parent Form-28 (CHQPF-28) and OSA-18 quality of life measures both pre- and postoperatively in the long and short term. Statistical analysis was performed using the unpaired Student t test, P value less than.05, and Spearman Rank coefficient. RESULTS: CHQ-PF28 scores were improved in the Physical Summary parameter in long-term follow-up. Psychosocial scores did not improve significantly. OSA-18 scores showed improvement in both the short-and long-term scores. Physical findings and symptoms did not impact scores in any domain. CONCLUSIONS: Quality of life in children with OSA does improve after adenotonsillectomy. Disease-specific clinometric instruments show improvement in domains affected by the disease process. However, instruments used to assess general quality of life may show physical improvement but not psychosocial. PMID- 14520089 TI - Natural history of untreated otitis media. AB - OBJECTIVE/HYPOTHESIS: Data from cohort studies and untreated groups in randomized controlled trials can be identified through systematic literature review and synthesized with meta-analysis to estimate natural history of acute otitis media (AOM) and otitis media with effusion (OME). STUDY DESIGN: Systematic literature review and meta-analysis. METHOD: Source articles were identified by MEDLINE search through August 2002 plus manual crosschecks of bibliographies and published meta-analyses. Data were abstracted independently by two investigators and combined with random effects meta-analysis to estimate spontaneous resolution, 95% confidence intervals (CI), and heterogeneity. Sensitivity analysis was performed. RESULTS: Sixty-three articles met inclusion criteria. AOM symptoms improved within 24 hours without antibiotics in 61% of children (95% CI, 50-72%), rising to 80% by 2 to 3 days (95% CI, 69-90%). Suppurative complications were comparable if antibiotics were withheld (0.12%) or provided (0.24%). Children entered recurrent AOM trials with a mean rate of 5.5 or more annual episodes but averaged only 2.8 annual episodes while on placebo (95% CI, 2.2 3.4). No AOM episodes occurred in 41%, and only 17% remained otitis prone (3 or more episodes). OME after untreated AOM had 59% resolution by 1 month (95% CI, 50 68%) and 74% resolution by 3 months (95% CI, 68-80%). OME of unknown duration had 28% spontaneous resolution by 3 months (95%, CI 14-41%), rising to 42% by 6 months (95% CI, 35-49%). In contrast, chronic OME had only 26% resolution by 6 months and 33% resolution by 1 year. CONCLUSIONS: The natural history of otitis media is very favorable. Combined estimates of spontaneous resolution provide a benchmark against which to judge new or established interventions. The need for surgery in children with recurrent AOM or chronic OME should be balanced against the likelihood of timely spontaneous resolution and the potential risk of learning, language, or other adverse sequelae from persistent middle ear effusion. Further research is needed to identify prognostic factors that can target children unlikely to improve spontaneously for earlier intervention. PMID- 14520090 TI - Autonomic related vertigo. AB - OBJECTIVES: To demonstrate clinical evidence that vestibular symptomatology can occur in relation to autonomic dysfunction. Characterization of clinical findings and treatment response would then allow autonomic related vertigo to be differentiated from other vertiginous conditions that present in a like manner. STUDY DESIGN: This was a retrospective review of 113 patients that described symptoms consistent with spontaneous, rotational vertigo and autonomic dysfunction. METHODS: Vestibular, otologic, and autonomic symptoms are presented along with the results of audiologic, orthostatic, and autonomic testing. Medical management included fluid loading, dietary changes, exercise, and patient education. Treatment results were analyzed according to the effectiveness in control of vestibular and otologic symptoms. Results were compared with a control group that demonstrated a similar vestibular and otologic presentation without autonomic symptomatology. RESULTS: All patients described spontaneous, rotational vertigo, with complete or substantial vertigo control obtained in 93 (85%) of 110 patients. Postural vertigo and distinct lightheadedness were also documented in 53% and 97% of cases, respectively. Vertigo failed to improve or worsened with prior treatment of low sodium diet or diuretic in 53 (91%) of 58 cases. Vertigo improvement was subsequently achieved in 48 (86%) of 56 cases with an autonomic treatment regimen. Long-term vertigo control was obtained in 56 (88%) of 64 patients followed for at least 18 months. Tinnitus was reported in 97 (86%) patients, aural fullness in 93 (82%) patients, and subjective hearing loss (HL) in 46 (41%) of 111 cases. Bilateral tinnitus and aural fullness occurred in 65% and 63%, respectively. Tinnitus improved with treatment in 56 (67%) of 84 patients, whereas aural fullness improved in 59 (74%) of 80 patients. Autonomic symptoms included palpitations in 103 (91%) patients, chronic fatigue in 102 (90%) patients, cold extremities in 91 (81%) patients, and previous fainting in 72 (64%) patients. A history of mitral valve prolapse was documented in 51 (45%) of cases and demonstrated with echocardiogram in 68 (93%) of the 73 patients tested. Audiologic testing was normal in 104 (95%) of 109 patients, and electrocochleography was abnormal in 42 (40%) of 105 patients. Orthostatic blood pressure and heart rate testing met the criteria for orthostatic hypotension in 16 (15%) of 104 patients. Autonomic testing was obtained in 34 cases, with orthostatic intolerance demonstrated in 33 (97%) patients and orthostatic hypotension demonstrated in 13 (38%) patients. Overall, orthostatic hypotension was documented through combined testing results in 23 (21%) of 107 patients. Vertigo was reproduced during autonomic testing in 17 (77%) of 22 patients, and otologic symptoms were reproduce in 9 (47%) of 19 patients. Comparison of the study population with a control group without autonomic symptoms revealed statistically significant differences in orthostatic testing and treatment results. There was no statistical difference noted in findings between patients of this study that demonstrated or failed to demonstrate orthostatic hypotension. CONCLUSIONS: There is a subgroup of patients with spontaneous vertigo who also demonstrate symptoms and findings consistent with poor autonomic regulation. These patients report vertigo improvement with a treatment strategy that aims to improve autonomic dysfunction through expansion of effective circulating volume. Clinical findings and treatment results of this study suggest an underlying autonomic influence in the production of vertigo and otologic symptoms. PMID- 14520091 TI - Presbycusis: a human temporal bone study of individuals with flat audiometric patterns of hearing loss using a new method to quantify stria vascularis volume. AB - OBJECTIVE: The purpose of this study was to determine the prevalence of stria vascularis atrophy in individuals with presbycusis and flat audiometric patterns of hearing loss. Individuals with presbycusis have historically been categorized by the shape of their audiograms, and flat audiometric thresholds have been reported to be associated with atrophy of the stria vascularis. Stria vascularis volume was not measured in these studies. STUDY DESIGN: Retrospective case review. METHODS: Archival human temporal bones from individuals with presbycusis were selected on the basis of strict audiometric criteria for flat audiometric thresholds. Six temporal bones that met these criteria were identified and compared with 10 temporal bones in individuals with normal hearing. A unique quantitative method was developed to measure the stria vascularis volume in these temporal bones. The hair cell and spiral ganglion cell populations also were quantitatively evaluated. RESULTS: Only one of the six individuals with presbycusis and flat audiometric thresholds had significant atrophy of the stria vascularis. This individual with stria vascularis atrophy also had reduced inner hair cell, outer hair cell, and ganglion cell populations. Three of the individuals with presbycusis had spiral ganglion cell loss, three individuals had inner hair cell loss, and all six individuals had outer hair cell loss. CONCLUSIONS: The results of this investigation suggest that individuals with presbycusis and flat audiometric patterns of hearing loss infrequently have stria vascularis atrophy. Outer hair cell loss alone or in combination with inner hair cell or ganglion cell loss may be the cause of flat audiometric thresholds in individuals with presbycusis. PMID- 14520093 TI - Outcome analysis of endoscopic sinus surgery in patients with nasal polyps and asthma. AB - OBJECTIVE: To investigate the efficacy of endoscopic sinus surgery (ESS) in the management of chronic sinusitis and asthma in patients with nasal polyps and steroid-dependent asthma. STUDY DESIGN: Retrospective chart review. METHODS: The study included 17 patients who underwent ESS with nasal polyps, steroid-dependent asthma with or without aspirin sensitivity and a minimum of 1 year postoperative follow-up. Nine patients were ASA sensitive, and eight patients were ASA tolerant. Chronic sinusitis and asthma were evaluated using subjective (patient complaints) and objective (computed tomography scans, pulmonary function tests, steroid doses) criteria. Preoperative data were compared with data obtained 12 to 18 months postESS. Tissue samples were graded for degree of inflammation and edema. RESULTS: Thirteen of the 17 (76.5%) patients reported improved clinical symptoms postESS. The postoperative Lund-Mackay scores were statistically lower for the 17 patients (P <.0001). The group experienced improvement in postoperative forced expiratory volume at 1 second (FEV1) (P <.014). Twelve of 17 (70.6%) experienced reduction in systemic steroid usage (P <.048). The ASA sensitive patients did not have a statistical improvement in postoperative FEV1 (P >.08) and sinonasal symptoms (P >.16) compared with the ASA tolerant group. Polyp tissue from the ASA sensitive patients demonstrated more edema and more inflammation on average than ASA tolerant polyps, but the results were not statistically significant. CONCLUSION: ESS demonstrates a beneficial effect on the sinonasal and asthma symptomatology in patients with nasal polyps and asthma using objective measures. Subset of aspirin-tolerant patients have statistically better outcome for sinonasal symptoms and pulmonary function testing than aspirin sensitive patients. PMID- 14520092 TI - Development of the Bowman-Birk inhibitor for oral cancer chemoprevention and analysis of Neu immunohistochemical staining intensity with Bowman-Birk inhibitor concentrate treatment. AB - OBJECTIVES/HYPOTHESIS: Cancer chemoprevention is a rapidly evolving approach to reverse or inhibit carcinogenesis, and there is active interest in development of effective chemopreventive agents against head and neck cancers. The retinoids are archetypal chemopreventive agents for oral premalignant lesions. They have significant clinical effect, but widespread use is limited by significant clinical toxicity. The Bowman-Birk Inhibitor is one of several nontoxic compounds exhibiting both potent anticarcinogenic activity and minimal toxicity. The purposes of the study were to summarize the preclinical and clinical development of Bowman-Birk Inhibitor and a Bowman-Birk Inhibitor concentrate against oral premalignant lesions and to evaluate Neu immunohistochemical staining intensity for lesions and simultaneously obtained biopsy specimens of normal-appearing mucosa from the Phase IIa Bowman-Birk Inhibitor concentrate oral leukoplakia chemoprevention trial. STUDY DESIGN: Part I is a selected literature review. Part II is a retrospective analysis of pathological specimens prospectively obtained from the Phase IIa clinical trial of Bowman-Birk Inhibitor concentrate. METHODS: Thirty-two sets of biopsy specimens from lesions and uninvolved oral mucosa before and after treatment with Bowman-Birk Inhibitor concentrate in doses ranging from 200 to 1066 chymotrypsin inhibitory units were examined in blinded fashion for Neu immunohistochemical staining intensity using the 3B-5 monoclonal antibody. Staining intensity scores among the lesion and control biopsy specimens before and after Bowman-Birk Inhibitor concentrate treatment were analyzed and compared with previously obtained values for serum Neu, oral mucosal cell Neu, protease activity, and clinical response to treatment. RESULTS: Mean Neu staining score was significantly higher in lesions compared with uninvolved mucosa (P <.001). Pretreatment staining scores for biopsy specimens of lesions and control biopsy specimens of normal-appearing tissues were correlated (Spearman correlation coefficient [r] = 0.375, P =.045), but no correlation between lesion and control biopsy specimen scores was evident after treatment. The change in Neu staining score with Bowman-Birk Inhibitor concentrate treatment in control site biopsy specimens demonstrated an inverse relationship of change in lesion area with Bowman-Birk Inhibitor concentrate treatment (Spearman r = -0.493, P <.007). CONCLUSION: Bowman-Birk Inhibitor concentrate shows promise to become an effective nontoxic chemopreventive agent based on results of extensive preclinical studies, and Phase I and Phase IIa clinical trials. Bowman-Birk Inhibitor concentrate has dose-related clinical activity against oral leukoplakia and modulates levels of Neu and protease activity. The current investigation identified increased Neu staining intensity in hyperplastic lesions compared with simultaneously obtained biopsy specimens of normal-appearing mucosa both before and after Bowman-Birk Inhibitor concentrate treatment. This finding supports prior observations that increased Neu expression is present in a subset of oral premalignant lesions and head and neck cancers. The trend of increased Neu staining score in control biopsy tissues of subjects exhibiting decreased lesion area following Bowman-Birk Inhibitor concentrate treatment raises questions about the mechanisms of Bowman-Birk Inhibitor concentrate action. One possible explanation is that Bowman-Birk Inhibitor stabilizes the extracellular domain of Neu, thereby preventing receptor truncation and internalization. Further study of modulation of Neu and protease activity by Bowman-Birk Inhibitor concentrate treatment may provide insights into the role of proteases and protease inhibitors in oral premalignant lesions and the mechanisms underlying Bowman-Birk Inhibitor concentrate effects. A Phase IIb randomized, placebo-controlled clinical trial to determine the clinical effectiveness of Bowman-Birk Inhibitor concentrate and further evaluate these candidate biomarkers is under way. PMID- 14520094 TI - Contralateral suppression of distortion-product otoacoustic emissions declines with age: a comparison of findings in CBA mice with human listeners. AB - OBJECTIVES/HYPOTHESIS: The auditory efferent system plays presumed roles in enhancing signals in noise, maintaining the cochlea for optimal acoustic signal processing, and may have a protective role in preserving auditory function in the face of ototoxic events. The objective of the study was to measure age-related changes of the medial olivocochlear efferent system in mice by comparing distortion-product otoacoustic emissions generated with and without contralateral white noise stimulation. Consistent with prior work, distortion-product otoacoustic emissions were typically reduced in magnitude when white noise was presented to the contralateral ear. This contralateral suppression is attributed to activation of the medial olivocochlear efferent system, which has an inhibitory effect on the cochlear hair cell system. By studying contralateral suppression on cochlear output in subjects of different ages, it is possible to describe aging effects on the medial olivocochlear system. STUDY DESIGN: CBA mice were divided into three age groups: young adult, middle-aged, and old-aged animals (21, 13, and 22 animals per group, respectively), and auditory brainstem responses were obtained before distortion-product otoacoustic emission testing to assess overall hearing abilities. METHODS: 2f1-f2 distortion-product otoacoustic emission recordings were obtained from individual subjects (anesthetized with ketamine/xylazine) in each age group under two conditions: 1) in quiet and 2) in the presence of a contralaterally applied wideband noise. RESULTS: Principal findings were that distortion-product otoacoustic emission levels decreased with age for mice in a way similar to humans, when correcting for the absolute difference in life spans. In addition, contralateral suppression declined in middle-aged and old-aged groups relative to the young adults for mice in a manner similar to humans. The contralateral suppression decline at low frequencies preceded that of the decline in distortion-product otoacoustic emissions with age. CONCLUSION: Functional decline of the medial olivocochlear efferent system with age precedes outer hair cell degeneration. Loss of medial olivocochlear suppressive function may play a role in the development of presbycusis in both clinical cases and animal models. PMID- 14520095 TI - Which comes first? Psychogenic dizziness versus otogenic anxiety. AB - SUMMARY: OBJECTIVE To investigate the hypotheses that physical neurotologic conditions may trigger anxiety disorders (otogenic pattern of illness), that psychiatric disorders may produce dizziness (psychogenic pattern), and that risk factors for these syndromes may be identified. STUDY DESIGN Retrospective review of all patients (N = 132) treated at a tertiary care balance center from 1998 to 2002 for psychogenic dizziness with or without physical neurotologic illnesses. METHODS All patients underwent comprehensive neurotologic and psychiatric evaluations with attention to the longitudinal course of symptoms and risk factors for psychopathology. Patients were grouped according to the condition first causing dizziness. Risk factors were compared across groups. RESULTS Three equally prevalent patterns of illness were found: anxiety disorders as the sole cause of dizziness (33% of cases), neurotologic conditions exacerbating preexisting psychiatric disorders (34%), and neurotologic conditions triggering new anxiety or depressive disorders (33%). Panic disorder and agoraphobia were significantly more prevalent than less severe phobias in the first two groups, whereas the opposite pattern existed in the third group (P <.0001). More patients in the first two groups had risk factors for anxiety disorders (P <.05). Depression was not a primary cause of dizziness in any patient. Vestibular neuronitis, benign paroxysmal positional vertigo, and migraine were the most common neurotologic conditions. CONCLUSIONS These data support the hypothesis that physical neurotologic conditions may trigger psychopathology as often as primary anxiety disorders cause dizziness. A third pattern appears to be equally common wherein physical neurotologic conditions exacerbate preexisting psychiatric illnesses. Individuals at risk for anxiety disorders may be more likely to have primary psychopathology. PMID- 14520096 TI - Tutorials in clinical research: VII. Understanding comparative statistics (contrast)--part B: application of T-test, Mann-Whitney U, and chi-square. AB - OBJECTIVE: This tutorial on comparative statistics has been written in two complementary segments. The first paper (part A) focused on explaining the general concepts of the null hypothesis and statistical significance. This second article (part B) addresses the application of three specific statistical tests. These two articles should be read sequentially and the first article should be available for reference while one reads the second. STUDY DESIGN: Tutorial. METHODS: The authors met weekly for 10 months to discuss clinical research articles and the applied statistics. The difficulty was not the material but the effort to make it easy to read and as short as possible. RESULTS: The article discusses the application of three common statistical indexes of contrast, chi2, Mann-Whitney U, and Student t-test and other concepts, such as sample size, degrees of freedom, errors, power, and confidence intervals. CONCLUSIONS: Statistical tests generate a number known as a statistic (chi2, U, t), which is sometimes called a "critical ratio" because it helps us to make a decision. This number is then associated with a probability, or P value. Sample size is a crucial element in the initial design of a research project and in the subsequent ability of the results to show statistical significance if the difference is clinically important. The example data used in this paper demonstrate the application of the three specific tests and illustrate the effect of sample size on the results. PMID- 14520097 TI - Combining acoustic and electrical hearing. AB - OBJECTIVES/HYPOTHESIS: The concept of combining electrical stimulation for high frequency sound with acoustic hearing for low-frequency information was tested. In addition, whether residual hearing can be preserved when an electrode is placed into the inner ear up to 10 mm and whether place of electrical stimulation influences speech perception were tested. STUDY DESIGN: A single-subject clinical trial design was employed. METHODS: Six postlingual adults with severe high frequency hearing impairment were recruited to participate in the study. A new six-channel cochlear implant was designed for the clinical trial. The intracochlear electrodes were either 6 or 10 mm in length based on a Nucleus CI 24 multichannel implant. Monosyllabic word understanding and consonant identification testing in a recorded sound-only condition were used to assess changes in speech perception. Follow-up was greater than 12 months. RESULTS: Acoustic hearing was preserved in all six subjects (n = 3, 6-mm electrodes; n = 3, 10-mm electrodes). Preoperative monosyllabic word and sentence scores were unchanged in all subjects following implantation. A 30% to 40% improvement in consonant recognition occurred with the 10-mm electrode. The subjects with 10-mm electrodes were able to understand 83% to 90% of the monosyllabic words using the implant plus binaural hearing aids. Scores were more than doubled when compared with preoperative scores with hearing aids only. CONCLUSION: The human ear has the capability to integrate both acoustic and high-frequency electrically processed speech information. Placement of a short, 10-mm electrode does not appear to damage residual low-frequency inner ear hair cell function, interfere with the micro mechanics of normal cochlear vibration, or decrease residual speech perception. The improvement in speech recognition was due primarily to the increased perception of higher-frequency consonantal speech cues, and this improvement took several months to become apparent. Such a device can provide a substantial benefit in speech understanding to individuals with severe high frequency hearing loss, while still maintaining the benefits of the residual lower-frequency acoustic hearing. The position of the electrode and the place of frequency information within the cochlea were shown to be important factors in the success of such a device. PMID- 14520098 TI - Ossicular chain reconstruction: titanium versus plastipore. AB - OBJECTIVE: To compare the complication rate and hearing results of a new, lightweight, titanium ossicular replacement prosthesis with Plastipore prostheses (Xomed, Jacksonville, FL). STUDY DESIGN: Retrospective. METHODS: Charts were reviewed for type of operation, type of prosthesis used, extrusion rate, prostheses failure rate, and hearing thresholds at multiple frequencies and at multiple follow-up points. The dependant variable for hearing results was the four-frequency average air-bone gap. RESULTS: There were 84 patients undergoing tympanoplasty with the Plastipore prosthesis and 53 with the titanium. There was one extrusion in the titanium group. There was an additional single incidence of prosthesis failure in the titanium group. Overall hearing results were comparable with an air-bone gap average of 19.3 dB in the Plastipore group compared with the titanium group with an air-bone gap of 22.0 dB (P =.08). Sixty percent of patients had a postoperative air-bone gap of 20 dB or less in the Plastipore group. In the titanium group, 45.3% achieved a 20 dB or less postoperative air bone gap. Plastipore had a lower air-bone gap than the titanium when a canal wall up operation was performed (17.8 vs. 23.9 dB) and tended toward a lower air-bone gap when a total ossicular prosthesis was needed (22 vs. 27 dB) (P <.07). CONCLUSION: The titanium prosthesis is a new ossicular replacement prosthesis that provides excellent visualization during insertion and provides hearing results that are comparable with Plastipore. The Plastipore prosthesis performed better in canal wall up mastoidectomy situations and tended toward better performance when a total ossicular replacement prosthesis was needed. PMID- 14520099 TI - Cost-effectiveness of intraoperative facial nerve monitoring in middle ear or mastoid surgery. AB - OBJECTIVES: Despite the utility of intraoperative facial nerve monitoring in reducing the risk of iatrogenic facial nerve injury during neurotologic surgery, its routine use during primary or revision surgery remains controversial. One of the major barriers to its acceptance is cost. This study evaluates the cost effectiveness of intraoperative facial nerve monitoring during middle ear or mastoid surgery. RESEARCH DESIGN/METHODS: A simple decision analytic cost effectiveness model and a societal approach were used to evaluate three cohorts of individuals who received (1) intraoperative facial nerve monitoring for both primary and revision middle ear or mastoid surgeries, or (2) facial nerve monitoring for revision surgeries only, or (3) no monitoring for any middle ear or mastoid surgeries. RESULTS: Our results strongly favored the use of intraoperative facial nerve monitoring in all patients undergoing middle ear or mastoid surgery, adding about $222.73 to $528.00 US dollars to the total cost. The strategy to monitor primary and revision surgeries had the greatest effectiveness and lowest cost, with an average quality-adjusted life-year (QALY) of 45.68 at an average cost of $238 US dollars. Facial nerve monitoring in revision patients only had similar QALYs (45.67) and higher costs ($292.1). Finally, the strategy not to monitor had the lowest QALY (45.65) and highest cost ($449.8). The analysis was robust across a wide range of changes in both costs and probabilities. CONCLUSIONS: Facial nerve monitoring is cost-effective, and its routine use should be adopted to reduce the risk of iatrogenic facial nerve injury during otologic surgery. PMID- 14520100 TI - Objective vestibular tests as outcome measures in head injury patients. AB - HYPOTHESIS: Dynamic visual acuity testing (DVAT) and the Dizziness Handicap Index (DHI) can be used as reliable outcome measures in patients after head injury. BACKGROUND: Balance disorders are a significant disability after mild traumatic brain injury (TBI). Assessing when individuals can perform activities of daily living, return to work, and begin to play sports can be difficult to determine. Objective outcome measures that correlate with successful life skills can be useful in managing these patients. METHODS: Fifty-three active duty individuals who suffered mild TBI underwent weekly DVAT testing and were administered a weekly DHI. Results in this group were compared with 46 control subjects who had not experienced TBI. In addition, weekly scores were compared with the patient's functional level, time to return to work, and time to perform all job related activities. RESULTS: Individuals with TBI showed an overall increase of 42% in DVAT function over the first 4 weeks of testing, whereas controls showed an 8% increase. Concurrently, individuals with TBI averaged a 18-point improvement in DHI function, whereas control subjects showed no significant change in this score. Improvement in DVAT and DHI function correlated closely at the 1-week time point. Improvement in the patient's cognitive function, ability to return to activities of daily living, and ability to return to work gradually improved continuously. CONCLUSION: DVAT and the DHI can be used as reliable outcome measures in evaluating the progress of patients with balance disorders associated with TBI. These measures allow providers to make more reliable recommendations regarding such activities as returning to work. PMID- 14520101 TI - A tympanometric comparison of tympanoplasty with cartilage palisades or fascia after surgery for tensa cholesteatoma in children. AB - OBJECTIVE: The aim of the present study is to compare the tympanometric and functional findings of cartilage palisade tympanoplasty with those of tympanoplasty with temporalis fascia grafting after one-stage surgery in children with tensa cholesteatoma. STUDY DESIGN: In children aged 5 to 15 years (mean, 9.5 years) with tensa cholesteatoma, cartilage palisade tympanoplasty was performed in 32 ears and fascia tympanoplasty in 29 ears. Tympanometry was performed in 31 ears with cartilage palisade tympanoplasty (the palisade group) and 28 ears with fascia tympanoplasty (the fascia group) at follow-up a median of 48 months after surgical removal of the cholesteatoma (range, 3-75 months). MAIN OUTCOME MEASURES: Postoperative perforations, tympanometric parameters (tympanogram type, compliance, tympanometric width), and hearing. RESULTS: All postoperative re perforations occurred in the fascia group. However, there was no difference between the palisade group and the fascia group in terms of tympanometric parameters, nor between the two groups when dividing into tensa retraction and sinus cholesteatoma subgroups. When comparing the types of tympanoplasty, the type II group reconstructed with cartilage palisades contained a higher number of ears with a normal compliance (0.2-1.6 mL) but a higher number of ears with a tympanometric width of more than 150 mmH2O. Regarding the number of ears with normal tympanometries, there was no significant difference between the palisade and the fascia group. The late functional hearing results were better in ears reconstructed with cartilage palisades, with functional success in 71%, compared with 54% in the group with fascia grafting. This difference in functional results was particularly pronounced in ears with an abnormal tympanogram (68% vs. 29%). CONCLUSION: Compared to fascia grafting, the present study showed better late functional hearing results after drum reconstruction using cartilage palisades, despite comparable tympanometric findings. Cartilage palisade reconstruction seems to provide better functional results, especially in ears with a poor tubal function, which is the common situation after cholesteatoma surgery. PMID- 14520102 TI - Connexin 26 mutations and nonsyndromic hearing impairment in northern Finland. AB - OBJECTIVE: The aims of the present study were to evaluate the role of the gap junction protein beta-2 gene (GJB2), encoding connexin 26 (Cx26), in children with moderate to profound prelingual nonsyndromic sensorineural hearing impairment (HI) and to investigate the carrier frequencies of the GJB2 gene mutations in a control population in Northern Finland. METHODS: Mutation analysis was performed by direct sequencing and carrier detection by conformation sensitive gel electrophoresis further confirmed by direct sequencing. RESULTS: Cx26 mutations were found in 15 of 71 (21.1%) (67 families) children with HI. Homozygosity for the mutation 35delG was shown to be the cause of HI in 13 of 15 (86.7%) children. Homozygosity for the M34T genotype was found in one child, and compound heterozygosity for the M34T/V37I genotype was found in another. Five families of those with suspected familial HI (29.4%) and six families out of those with sporadic HI (12.0%) had a homozygous or compound heterozygous mutation. The carrier frequency for the mutation 35delG was 1 of 78 (4 of 313) and that for the M34T was 1 of 26 (12 of 313). CONCLUSION: 35delG/35delG genotype was found to be a significant cause of moderate to profound prelingual nonsyndromic sensorineural HI in Northern Finland. M34T/M34T genotype was seen in only one child, but the carrier frequency of the M34T allele was about three times higher than that of the 35delG mutation. PMID- 14520103 TI - Protease inhibitors alpha1-antitrypsin and ilomastat are not ototoxic in the chinchilla. AB - OBJECTIVES: Proteases of both the serine and the metalloprotease families have been shown to play a role in the pathogenesis of otitis media. Inhibitors of proteases from each of these families have been shown to beneficially impact disease progression in a number of related chronic inflammatory conditions. The purpose of this study was to assess the safety of protease inhibitors when instilled into the middle ear, with a view to their potential use in the treatment of human otitis media. STUDY DESIGN: Prospective, randomized, controlled trial in the chinchilla model. METHODS: After completing baseline auditory testing and bilateral transpalatal obstruction of the Eustachian tube, chinchillas received weekly transbullar injections of protease inhibitor (alpha1 antitrypsin, ilomastat, or both), vehicle, or saline. After 1 month, hearing was tested and the animals were sacrificed. Temporal bone histopathologic examination was performed. RESULTS: All treatment groups demonstrated a statistically insignificant average loss in long-term hearing (0 dB) for all measures using clicks and tones (P >.15 for all conditions). All treatment groups were statistically insignificantly different from one another (P =.5625). Histopathologic examination revealed no significant inner ear changes. CONCLUSIONS: Protease inhibitors that are currently under study in animal models and humans for the treatment of inflammatory diseases that are related to imbalances between protease and protease inhibitor have no significant toxic effect on the inner ear of chinchillas. These findings support the safety of further clinical trials using these inhibitors to treat middle ear inflammation. PMID- 14520104 TI - Antibodies to HSP-70 in normal donors and autoimmune hearing loss patients. AB - OBJECTIVE: To evaluate serum antibody to heat shock protein (HSP) 70 as a marker for autoimmune sensorineural hearing loss (AISNHL). DESIGN: Sera from 20 patients with rapidly progressive sensorineural HL and 20 control volunteers without HL were tested for antibody reactivity against multiple HSP 70 substrates. Substrates included recombinant human HSP (rHuHSP) 72, purified bovine brain heat shock cognate (HSC) 73 and HSP 72, as well as heat-shocked and non-heat-shocked protein extracts from bovine kidney (MDBK) cells. All serum donors were previously tested for antibody to guinea pig inner ear supporting cells; 17 of 20 patients but none (0 of 20) of the controls were positive. METHODS: Sera were tested using Western blots. RESULTS: Reactivity with rHuHSP 70 was observed in 16 patients and 17 controls. Similarly, 15 of 20 patients and 17 of 20 controls stained for both HSP 72 and HSC 73 from the bovine brain. When tested against the heat-shock-induced and control MDBK extracts, six patients and nine controls had greater reactivity with the induced HSP 72. CONCLUSION: The frequency of antibodies to HSP substrates did not differ in patients and controls. Prior studies reported that HSP 72 is the 68 kD antigen commonly detected by AISNHL sera. However, we show that HSP 72 antibodies are no more prevalent in patients than in normal controls. Thus, it is unlikely that the 68 kD protein is HSP 72. Therefore, HSPs are not appropriate substrates for serodiagnosis of AISNHL. PMID- 14520105 TI - Safe tracheostomy for patients with severe acute respiratory syndrome. AB - OBJECTIVES/HYPOTHESIS: Severe acute respiratory syndrome (SARS) caused by coronavirus has become an epidemic affecting many regions worldwide. Fourteen percent to 20% of patients require endotracheal intubation and ventilator support. Some of these patients may require tracheostomy subsequently. This procedure, when performed without protection, may lead to infection of the medical and nursing staff taking care of the patient. STUDY DESIGN: Based on clinical information of three patients. METHODS: The authors carried out an emergency tracheostomy and changed the tracheostomy tube for one patient and performed elective tracheostomy in another two patients. RESULTS: No medical or nursing staff member was infected after carrying out the procedure while taking all the precautions and wearing the appropriate protective apparel. CONCLUSION: The authors have prepared guidelines for performing a safe tracheostomy under both elective and emergency conditions. Surgeons who might be involved in performing the tracheostomy should become familiar with these guidelines and the appropriate protective apparel. PMID- 14520106 TI - Nasopharyngeal flora and drug susceptibility in children with macrolide therapy. AB - OBJECTIVES/HYPOTHESIS: Low-dose, long-term administration of macrolides (macrolide therapy) has been used as an effective treatment for chronic respiratory tract diseases. The authors reported on the nasopharyngeal flora in children treated with macrolide therapy. STUDY DESIGN: Prospective study. METHODS: Nasopharyngeal cultures were obtained from 73 children with chronic rhinosinusitis and/or otitis media with effusion at the end of the low-dose administration of clarithromycin (macrolide group). As control subjects, 98 children with chronic rhinosinusitis and/or otitis media with effusion who were not given macrolides were also included in the study. The culture results were evaluated with respect to antimicrobial susceptibility patterns, risk factors for carriage of erythromycin-resistant Streptococcus pneumoniae, and the clinical efficacy of the therapy. RESULTS: The macrolide therapy did not have a significant effect on the incidence or the susceptibility patterns of potential pathogens except for Moraxella catarrhalis. Most of children in the macrolide group possessed a normal flora compared with the control children. The risk factors for carriage of erythromycin-resistant S pneumoniae were male gender in the macrolide group and age under 6 years and use of antimicrobial drugs other than macrolides in the control group. The clinical efficacy of the therapy was independent of carriage of erythromycin-resistant S pneumoniae. CONCLUSION: Macrolide therapy has little effect on carriage of drug-resistant pathogens, and the efficacy of the therapy depends on the anti-inflammatory effect of the drugs, which is independent of their antimicrobial effect. PMID- 14520107 TI - Multilevel temperature-controlled radiofrequency therapy of soft palate, base of tongue, and tonsils in adults with obstructive sleep apnea. AB - OBJECTIVES/HYPOTHESIS: The concept of two-level pharyngeal collapse in patients with obstructive sleep apnea is too simplified. Aggressive multilevel surgeries addressing several airway segments, including skeletal surgery, demonstrate improved success rates. STUDY DESIGN: The study aimed to evaluate the safety and efficacy of multilevel radiofrequency application to soft palate, tonsils, and base of tongue in 16 white patients (mean age, 56.9 +/- 11.1 y; mean body mass index, 27.3 +/- 2.6 kg/m2) with obstructive sleep apnea. There was one dropout. Therapeutic effects after one treatment session were assessed 20.6 +/- 12.6 weeks postoperatively. Treatment outcome measurements were based on Epworth Sleepiness Scale, Likert scales, and polysomnography. METHODS: Every patient received 16 treatment sites with a total dose of 9750 J radiofrequency energy into soft palate, base of tongue, and tonsils. Success was defined as respiratory disturbance index equal to or less than 20 or at least 50% improvement if baseline respiratory disturbance index was less than 20. Statistical analysis was determined with the Spearman rank test. RESULTS: Mean score on Epworth Sleepiness Scale decreased from 11.1 to 8.2 (P =.0001). Of the patients, 53.3% reported improvement of their daytime sleepiness. Snoring was assessed with 10-point Lickert scale, and score decreased from 7.5 to 4.9 (P =.08). Mean respiratory disturbance index decreased from 32.6 +/- 17.4 to 22.0 +/- 15.0 (P =.003). By our definition of success, 5 of 15 patients (33%) have been treated successfully surgically and 4 of 15 (27%) had remarkable improvement after one treatment session; 1 patient (6.6%) demonstrated deterioration. There were two adverse effects, one superficial ulceration of the soft palate and one unilateral tonsillar abscess formation, with an overall complication rate of 13.3% for our patients and 0.41% for all treatment sites (n = 240). CONCLUSION: Moderate to severe obstructive sleep apnea usually requires multilevel pharyngeal surgery. Radiofrequency offers the potential of altering the upper airway on different sites. PMID- 14520108 TI - Tissue engineered muscle implantation for tongue reconstruction: a preliminary report. AB - OBJECTIVES/HYPOTHESIS: Because current tongue reconstructive methods introduce adynamic, variably sensate tissue into the mouth, the critical functions of the tongue in articulation and deglutition may be compromised. The objective of this work was to introduce a combination of myoblasts and scaffolding material into rat hemiglossectomy defects and to examine the extent of neomuscle formation in the reconstructed area, under the hypothesis that the presence of myoblasts leads to formation of new muscle. STUDY DESIGN: Randomized, prospective animal study. METHODS: Myoblasts were harvested from neonatal Lewis rats, and a growth factor enriched collagen gel was prepared. Syngeneic adult animals received either hemiglossectomy alone or reconstruction with one of four experimental reconstructive preparations: collagen gel alone, collagen gel with suspended myoblasts, the gel-cell combination in undifferentiated muscle construct form by way of tissue culture for 7 days in a preformed mold, or differentiated constructs, cultured in myoblast fusion medium. After 6 or 16 weeks, animal weight gain was recorded, animals were killed, and the tongues harvested. The tissue was examined histologically, and quality of the muscular regenerate was rated on a scale according to predefined criteria. RESULTS: Animals in all groups gained weight appropriately. In groups receiving hemiglossectomy alone or acellular (gel only) reconstruction, there was significant scarring and lack of neomuscle formation. In groups receiving myoblast transplantation, either by way of gel suspension or in the form of undifferentiated or differentiated constructs, muscle quality was superior to controls. CONCLUSIONS: Myoblast transplantation into hemiglossectomy defects appears to lead to new muscle formation and does not inhibit normal weight gain in animals after tongue implantation. PMID- 14520109 TI - Local production of Aspergillus fumigatus specific immunoglobulin E in nasal polyps. AB - BACKGROUND: Aspergillus spp. play a significant role in the etiology of immunoglobulin (Ig)E mediated allergic fungal sinusitis (AFS). It is unclear whether Aspergillus spp. are also involved in nasal polyps without the characteristic clinical features of AFS. OBJECTIVES: The frequency of Aspergillus spp. and Aspergillus-specific IgE in nasal lavages and serum of patients with severe nasal polyps (n = 33) without clinical features of AFS should be investigated. STUDY DESIGN: Prospective study. METHODS: An aliquot of nasal lavage fluid was treated with dithiothreitol and examined for Aspergillus fumigatus by culture and an Aspergillus-specific polymerase chain reaction (PCR) assay. An additional aliquot of nasal fluid and serum of the same patient were tested for specific IgE (Unicap, Pharmacia, Freiburg, Germany) to recombinant Aspergillus fumigatus allergen (rAspf) 1 to 6. RESULTS: All patients had negative skin prick tests for Aspergillus fumigatus. Four of 33 (12%) lavage samples were positive for Aspergillus spp. by PCR. In one of these samples, rAspf-specific IgE was detected but none in the serum. Nasal lavage and serum samples of the remaining 29 patients were negative for rAspf-specific IgE. CONCLUSIONS: Aspergillus spp. detection is rare in patients with severe nasal polyps without characteristic clinical features of AFS. Specific IgE in nasal secretions may be elevated in patients with negative skin prick tests and serum IgE. In these cases, immunologic mechanisms similar to AFS may be involved. Fungal etiology has been proposed to underlie severe nasal polyps in general. However, Aspergillus spp. seem not to play a significant role. PMID- 14520111 TI - Level of vocal folds as projected on the exterior thyroid cartilage. AB - OBJECTIVE: This study aims to identify the level of the vocal folds as projected on the exterior thyroid cartilage. STUDY DESIGN: Anatomic study of human cadaver larynges. METHODS: The study includes 83 fresh larynges harvested at autopsy from 62 male and 21 female cadavers. The larynges were excised and divided in the midline posteriorly. One needle was inserted at the level of anterior commissure from endolarynx, and the other was inserted at the thyroid ala just anterior to the vocal process along the superior surface of the right vocal cord. Measurements of vocal cord projections on the thyroid ala were done with a caliper. RESULTS: The mean value of the ratio of the distances from the superior thyroid notch to anterior commissure and the midline height from thyroid notch to the inferior border of thyroid cartilage was found to be 0.41 in males and 0.38 in females. No statistical differences were observed between these two groups (P =.062). We found that the distance from the anterior commissure to the inferior thyroid border in midline "c" was longer than the distance from the posterior border of the vocal cord to the inferior border of the inferior tubercle of the thyroid ala "d" in 44 (71%) males and in 7 (33%) females. On the other hand, "d" was longer than "c" in 8 (12.9%) males and in 8 (38.1%) females. These two distances were equal in 10 (16.1%) males and in 6 (28.6%) females. CONCLUSION: In this study, we found that the anterior commissure lies approximately at the juncture of the upper two fifths and lower three fifths of the midline height of thyroid cartilage in the majority of the larynges of the male and female cadavers. The position of the posterior border of the vocal cords was found to be at a lower level than anterior commissure in two thirds of males and in one third of females. This means that the vocal cords slope downward posteriorly in the majority of the larynges of the males. This may be one of the causes of failure of some type I thyroplasties. PMID- 14520110 TI - IL-11 and IL-17 expression in nasal polyps: relationship to collagen deposition and suppression by intranasal fluticasone propionate. AB - OBJECTIVES/HYPOTHESIS: Chronic hyperplastic sinusitis (CHS) with nasal polyps (NP) is characterized by extensive mucosal thickening, goblet cell hyperplasia, and subepithelial fibrosis. These features are described to be part of remodeling in the lower airways. The cytokines interleukin (IL)-11 and IL-17 are believed to play a role in lower airway remodeling, but there has been very little work so far examining these cytokines and their relationship to fibrosis in CHS/NP. The aims of this study were to examine the deposition of collagens types I, III, and V in CHS/NP, evaluate the relationship of collagen deposition to expression of IL 11 and IL-17, and to examine the effect of treatment with intranasal fluticasone on these features. STUDY DESIGN: Sixteen subjects were included in this double blind, placebo-controlled study. NP biopsies were obtained at the baseline and after 4 weeks of treatment with intranasal fluticasone propionate (FP, Flonase) or placebo. Normal control middle turbinate biopsies from eight nonallergic subjects without sinusitis were used as a control for cytokine and collagen expression. METHODS: Tissues were assessed for deposition of collagen types I, III, and V using immunocytochemistry. The expression of the cytokines IL-11 and IL-17 was examined by immunostaining or in situ hybridization. The pre- to posttreatment results were analyzed using paired t test, and the magnitude of changes were estimated using one-way analysis of variance (ANOVA) statistical test followed by least significance difference post hoc comparisons of means. RESULTS: Compared with normal control nasal turbinate tissues, collagen types I, III, and V were increased in all NP tissues, with a predominance of types III and V. Collagen deposition was most abundant in the submucosal connective tissue and in the basement membrane zone. FP treatment had no significant effect on deposition of any collagen type. Expression of IL-11 and IL-17 was also greatly increased in NP compared with control nasal turbinate tissues. IL-11 expression was observed in both inflammatory cells and the epithelium, whereas IL-17 expression was primarily associated with inflammatory cells. In the pretreatment NP, a correlation was found between the presence of IL-11 and collagen type I (r = 0.59, P =.02) and also between IL-17 and both CD4+ and CD8+ T lymphocytes (r = 0.52, P =.05; r = 0.60, P =.02, respectively). Treatment with FP significantly reduced IL-11 expression in subepithelial inflammatory cells and in the epithelial compartment. In contrast, although IL-17 expression was reduced by FP, this effect did not reach statistical significance. CONCLUSION: NP manifest an increased expression of collagen types III, V, and I and an increase in profibotic cytokines IL-11 and IL-17. A correlation exists between deposition of collagen type I and expression of IL-11, suggesting a possible role for IL-11 in NP remodeling. Collagen deposition was not reversed by FP treatment, whereas IL 11 expression was suppressed. These results are consistent with a partial insensitivity of NP to FP treatment but also suggest that longer-term treatment or perhaps earlier intervention with FP might reduce proinflammatory cytokine signals and ultimately have a beneficial effect in preventing airway remodeling in NP. PMID- 14520112 TI - Hemicricoidectomy as the primary diagnosis and treatment for cricoid chondrosarcomas. AB - OBJECTIVES/HYPOTHESIS: The objective was to present a new approach for the diagnosis and treatment of chondrosarcoma involving the cricoid cartilage. The technique involved an extramucosal resection of the ipsilateral half of the involved cricoid cartilage, providing enough tissue to be sent for pathological study, and resulted in good laryngeal function without jeopardizing patients' long-term survival. STUDY DESIGN: Retrospective study. METHODS: A retrospective study of eight patients who underwent hemicricoidectomy for cricoid chondrosarcoma was performed at the Center for Voice Disorders, Wake Forest University (Winston-Salem, NC). One patient was a woman and seven were men. The mean age at diagnosis was 64 years (age range, 53-72 y). RESULTS: All patients had a low-grade tumor. Primary treatment included hemicricoidectomy in all cases. In six (75%) of the patients, tracheotomy was required at the initial surgery. The mean time for decannulation was 3.2 months (range, 5 d-1 y). In four cases a second procedure was required because of recurrence. The second procedure included two total laryngectomies and two endoscopic carbon dioxide laser excisions. The mean follow-up time was 3 years (range, 2 mo-10 y). At the time of writing, six patients were alive without recurrence, one patient was alive with disease; and one patient had died of unrelated causes. CONCLUSION: The authors recommended unilateral hemicricoidectomy as the diagnosis and treatment of choice. This procedure allows sufficient tissue for histological study and provides good long-term breathing and phonatory function without compromising long-term survival. This procedure is appropriate for patients with 1) mobility of one vocal fold, 2) dysphonia or aphonia, and 3) an adequate subglottic airway. PMID- 14520113 TI - Total thyroidectomy for benign thyroid disease. AB - OBJECTIVES/HYPOTHESIS: The use of total thyroidectomy in thyroid cancer treatment is not unanimous, and it is even more controversial when this procedure is advocated for benign diseases. On the other hand, the complication risk may have an increase up to 20 times in repeat operations for recurrence. The objective of the study was to evaluate the use of total thyroidectomy in benign diseases, multinodular goiter, and Graves disease to justify the authors' preference. STUDY DESIGN: Retrospective study of use of total thyroidectomy in benign diseases. METHODS: Retrospective study of 1789 patients who underwent thyroidectomies from June 1990 to December 2000. Indication, extension of thyroidectomy, cancer incidence, and complications were analyzed. RESULTS: Total thyroidectomy was performed in 81.19% of 456 patients with nontoxic multinodular goiter, 93.93% of 33 with toxic multinodular goiter, 93.93% of 66 with recurrent multinodular goiter, and 49.18% of 122 with Graves disease. Thyroid cancer was found in 16.62%, 9.09%, 3.03% and 5.73% of patients, respectively. Transitory and permanent hypoparathyroidism, hematoma requiring surgical intervention, and transitory and permanent recurrent laryngeal nerve injury occurred in 12.27%, 1.61%, 0.26%, 1.88%, and 0.35% of the patients undergoing total thyroidectomy, respectively. Permanent complications of total thyroidectomy for nontoxic multinodular goiter and Graves disease were similar to nontotal thyroidectomy. Use of total thyroidectomy for nontoxic multinodular goiter increased from 53.33% of the patient to 81.19%, on average, with a concomitant increase of cancer diagnosis from 11.11% to 16,62%. The authors performed total thyroidectomy for all patients with Graves disease. CONCLUSION: Total thyroidectomy is the treatment of choice for multinodular goiter and thyroiditis, when there is bilateral gland involvement posterior to middle thyroid veins, and for Graves disease because it decreases the likelihood of future repeat operations for recurrent disease and thus the associated risks, when performed safely. PMID- 14520114 TI - Cutaneous head and neck squamous cell carcinoma metastatic to cervical lymph nodes (nonparotid): a better outcome with surgery and adjuvant radiotherapy. AB - OBJECTIVE: The aim of this study is to report on the experience of treating cutaneous squamous cell carcinoma (SCC) metastatic to cervical (nonparotid) lymph nodes at the Head and Neck Unit, Westmead Hospital, Sydney, Australia. STUDY DESIGN: Retrospective chart review. METHODS: Patients diagnosed with previously untreated metastatic cutaneous SCC to cervical lymph nodes (levels I-V) and treated with radiotherapy, surgery, or surgery and adjuvant radiotherapy were identified. Relapse and outcome was analyzed using Cox regression analysis. RESULTS: Between 1980 and 2000, 74 patients were treated with curative intent. There were 59 males and 15 females, with a median age of 66 (range 37-93) years. Median duration of follow up was 48 (range 12-187) months. Fifty-two were treated with neck dissection and radiotherapy, 13 with neck dissection alone, and 9 with only radiotherapy. Most patients (85%) had an identifiable index lesion. Level I (38%) and II (36%) lymph nodes were the most often involved. In total, 25 (34%) patients developed recurrent disease, predominantly locoregional (22 of 25). Median time to recurrence was 5.2 (2-34.3) months. Increasing nodal size (> or =3 cm) (P =.01), metastatic spread to multiple nodes (P =.05), and the presence of extranodal spread (P =.01) all predicted for worse survival. Patients undergoing combined modality treatment had a lower relapse rate (15%) and a significantly better disease free survival (P =.001) compared with single modality treatment. CONCLUSION: Metastatic cutaneous SCC is uncommon but potentially lethal. Surgery and adjuvant radiotherapy remain the best practice and provide the best chance of achieving locoregional control. PMID- 14520115 TI - Foresight and courage: Levi Cooper Lane, MD, a model head and neck surgeon. AB - OBJECTIVES: To detail the life and achievements of Levi Cooper Lane, MD, author of the first American textbook on surgery of the head and neck. METHODS: Literature search and analysis. RESULTS/CONCLUSIONS: While some of Lane's observations have been diluted by the passage of time, the lessons imparted by his life and work merit continued study by contemporary head and neck surgeons. PMID- 14520118 TI - Significant dose effect of carbamazepine on reduction of steady-state plasma concentration of haloperidol in schizophrenic patients. AB - A reduction in haloperidol concentration induced by carbamazepine coadministration has been consistently reported. However, the degree of this reduction is very different among individuals treated with various doses of carbamazepine. Thus, we investigated dose effect of carbamazepine on the steady state plasma concentration of haloperidol. Eleven excited schizophrenic inpatients, despite receiving haloperidol 12 mg/d, were treated with incremental doses of carbamazepine for 6 weeks (100, 300, 600 mg/d for 2 weeks each). Plasma drug concentrations were monitored together with clinical assessments before and after each phase of the 3 carbamazepine doses. Mean haloperidol concentrations during coadministration of carbamazepine 100, 300, and 600 mg/d were 75%, 39%, and 15%, respectively, of corresponding variables before carbamazepine coadministration. Negative linear correlations were observed between dose or plasma concentration of carbamazepine and the degree of reduction in haloperidol concentration. Mean carbamazepine dose and plasma carbamazepine concentrations at 50% reduction of haloperidol concentration were 240 mg/d and 3.5 microg/mL, respectively. Scores in total and excitement symptoms were significantly reduced after carbamazepine coadministration, whereas no changes were observed in other clinical symptoms or any of the subgroup side effects. Therefore, this study indicates that carbamazepine decreases plasma haloperidol concentration in a dose dependent or concentration-dependent manner, and that reduction in haloperidol concentration is apparent even at subtherapeutic dose of carbamazepine. PMID- 14520117 TI - Catechol-O-methyltransferase and monoamine oxidase A genotypes and drug response to conventional neuroleptics in schizophrenia. AB - Biogenic amine synthesis and degradation are involved in the pathogenesis of schizophrenia. Catechol-O-methyltransferase and monoamine oxidase enzymes are important agents in the metabolic inactivation of these neurotransmitters (ie, dopamine, serotonin, and norepinephrine). Functional polymorphism in the catechol O-methyltransferase and monoamine oxidase A genes causes variation in enzyme activities. We investigated the relationship of catechol-O-methyltransferase Val158Met and monoamine oxidase A promoter repeat polymorphism with response to conventional neuroleptic treatment in schizophrenia.Ninety-four schizophrenic patients formed 2 different study populations. The responders had experienced a fair and steady response to conventional neuroleptics. The nonresponders had failed to achieve an acceptable response to conventional neuroleptics. We also used a control population of 94 age-matched and gender-matched blood donors. Genotyping of the catechol-O-methyltransferase and monoamine oxidase A genes was performed by polymerase chain reaction.Forty-three percent of the nonresponders had a low activity catechol-O-methyltransferase genotype compared with 16% of the responders (P = 0.009). Monoamine oxidase A genotype alone did not differ significantly between the groups. Moreover, the risk of having both low-activity catechol-O-methyltransferase and monoamine oxidase A genotypes was over 6 times more common (odds ratio = 6.16, P = 0.03) in the nonresponders compared with responders. The whole population of patients with schizophrenia did not differ from the controls.The low-activity catechol-O-methyltransferase genotype may be associated with unsatisfactory drug response to conventional neuroleptics or alternatively be involved in a subset of schizophrenics. The role of monoamine oxidase A genotype seems to be additive in this respect. PMID- 14520119 TI - Influences of an anticholinergic antiparkinsonian drug, parkinsonism, and psychotic symptoms on cardiac autonomic function in schizophrenia. AB - The arguments against the use of anticholinergic antiparkinsonian drugs for neuroleptic-induced parkinsonism have been based, in part, on their autonomic side effects. Except for anecdotal case reports, there is little evidence that antiparkinsonian drugs are the main factor causing autonomic dysfunction in schizophrenic patients with parkinsonism. Therefore, in the current study, the separate influences of the anticholinergic antiparkinsonian drug (biperiden), parkinsonism, and psychotic symptoms on cardiac autonomic function were investigated in 48 patients with schizophrenia. Biperiden was discontinued in 33 patients with or without parkinsonism and commenced in 15 patients with parkinsonism. Their parkinsonism and psychotic symptoms were assessed using rating scales, and their cardiac autonomic functions were assessed using the mean R-R interval and 3 methods of analyzing heart rate variability both before and after the change in medication. Consequently, the cardiac autonomic function was not affected by biperiden or the change in parkinsonism. Cardiac vagal function decreased when psychotic symptoms were more pronounced, but cardiac sympathetic function did not show a significant change. Therefore, it appeared that psychotic symptoms played the predominant role in modifying the cardiac autonomic function, implying the existence of autonomic changes associated with cognitive processing and a possible relation between psychotic symptoms and autonomic symptoms in schizophrenia. PMID- 14520120 TI - Lack of efficacy of low doses of quetiapine addition in refractory obsessive compulsive disorder. AB - BACKGROUND: Some studies have shown that low dose of neuroleptic addition to ongoing antiobsessive treatment might be effective in treatment resistant obsessive-compulsive disorder (OCD). The primary purpose of this study was to evaluate the efficacy of low-dose quetiapine in clomipramine and serotonin reuptake inhibitor-refractory obsessive-compulsive disorder patients. METHOD: Eight obsessive-compulsive disorder patients who were resistant to treatment after 2 or more inadequate trials with clomipramine or serotonin reuptake inhibitor were admitted to the study. The patients were administrated on open trial of quetiapine at daily dose of 150 mg. Treatment response was assessed using the Yale-Brown Compulsive Scale and the Clinical Global Impressions Scale. RESULTS: Only 2 patients were partial responders, with Yale-Brown Compulsive Scale score decreases of 28% and 29%. CONCLUSION: The results of this preliminary open trial suggest that low dose of quetiapine may not be effective in treatment resistant obsessive-compulsive disorder patients. Larger placebo-controlled trials are needed to investigate the clinical efficacy of quetiapine addition at low doses to antiobsessive treatment in treatment-refractory obsessive-compulsive disorder. PMID- 14520121 TI - Thrombocytopenia during valproic acid treatment in young patients with new-onset bipolar disorder. AB - BACKGROUND: To evaluate whether valproic acid (VPA) can cause thrombocytopenia and impaired platelet function in young patients with new-onset bipolar disorder. METHODS: The authors studied 25 new-onset young bipolar patients. Platelet count, platelet aggregation, platelet release, and bleeding time were evaluated before beginning VPA treatment and at least after 10 months of treatment. The control group consisted of 20 sex-matched and age-matched subjects. Patients were started on VPA at a dose of 250 to 750 mg/d, given in divided doses. Mean dosage of VPA was 1137.5 +/- 241.1 mg/d. Mean VPA total plasma concentration was 61.1 +/- 20 g/mL. RESULTS: At baseline, no significant differences were observed for platelet count and function between the bipolar group and the control subjects. After 10 months, at the second evaluation, the platelet count was significantly lower in the bipolar patients than in the control subjects: 192.7 +/- 21.4/microL versus 289.8 +/- 23.9/microL; P < 0.0001. An important observation was that platelet counts were negatively correlated with VPA dose (r = -0.47; P = 0.05) and its plasma concentration (r = -0.50; P = 0.05). In the present study, the authors observed impairment in platelet release of ATP and aggregation that correlated with both VPA dosage and plasma levels. Bleeding times were also significantly longer in patients taking VPA compared with control subjects (P < 0.0001). CONCLUSION: Thrombocytopenia can appear after a few months of therapy and with plasma VPA levels within the therapeutic range. PMID- 14520122 TI - Effects of polymorphisms in CYP2D6, CYP2C9, and CYP2C19 on trimipramine pharmacokinetics. AB - Little is known about the impact of cytochrome P450 polymorphisms on the metabolism of trimipramine, which is still widely used as antidepressant due to its positive effect on sleep patterns. A single oral dose of 75 mg trimipramine was given to 42 healthy volunteers selected according to their CYP2D6, CYP2C19, and CYP2C9 genotypes. The reference group included 8 subjects with homozygous active wild-type genotypes of all 3 enzymes (EM). This group was compared with 7 intermediate (IM) with 1 and 7 poor metabolizers (PM) with zero active alleles of CYP2D6 and CYP2C19, respectively, and with 4 subjects with the genotype CYP2C9*3/*3. Pharmacokinetics of trimipramine and its demethylated metabolite strongly depended on the CYP2D6 genotype. Median oral clearance of trimipramine was 276 L/h (range 180-444) in the reference group but only 36 L/h (range 24-48) in CYP2D6 PMs (P < 0.001). These differences could only be explained by an effect of CYP genotypes on both parameters, systemic clearance and bioavailability, the latter being at least 3-fold higher in CYP2D6 PMs than in the reference group. The desmethyltrimipramine area under the concentration-time curve was 40-fold greater in CYP2D6 PMs than in the reference group (1.7 vs. 0.04 mg/L x h in EMs), but below the quantification limit in most carriers of deficiencies of CYP2C19 or CYP2C9. This indicates that both CYP2C enzymes contribute to the demethylation of desmethyltrimipramine and CYP2D6 to further metabolism. PMID- 14520124 TI - Dose-related pharmacokinetics and pharmacodynamics of valproate in the elderly. AB - Valproate exhibits a complex pharmacokinetic profile due to concentration dependent protein binding and clearance. It has been shown that the protein binding of valproate decreases as the serum concentration increases in a young adult population. Furthermore, the percentage of protein binding is lower in the elderly compared with young adults at comparable low therapeutic serum concentrations. The extent of valproate protein binding at higher concentrations in the elderly has not been described. Studies conducted in the elderly have found unbound valproate clearance to be decreased compared with younger adults, although these changes in clearance have not been evaluated at higher therapeutic serum concentrations. We evaluated the pharmacokinetics of valproate (protein binding and clearance) across a wide dosage range in the elderly and measured the impact of this on drug-related side effects using a single-blind within-subject study design in 6 healthy elderly volunteers (aged 65-76 years). Steady-state total and unbound serum valproate concentrations were assessed at 3 doses: 500, 1000, and 1500 mg/d. As doses and valproate serum concentrations increased, the unbound fraction (10.0%, 13.0%, 17.4%) and total clearance (4.8, 6.0, 6.7 mL/h/kg) increased, respectively. Unbound clearance decreased (49.4, 45.8, 39.4 mL/h/kg) with increasing valproate serum concentrations. Drug-induced CNS effects and nausea severity scores correlated with total and unbound serum valproate concentrations. Significant dose-dependent changes in valproate pharmacokinetics were observed in the elderly. PMID- 14520123 TI - Antidepressants and ejaculation: a double-blind, randomized, fixed-dose study with mirtazapine and paroxetine. AB - A double-blind, fixed-dose study in healthy men with lifelong early ejaculation was performed to evaluate potential differences in their effects on ejaculation latency, between clinically relevant doses of the selective serotonin reuptake inhibitor paroxetine and the noradrenergic and specific serotonergic antidepressant mirtazapine. Twenty-four men with an intravaginal ejaculation latency time (IELT) less than 1 minute were randomly assigned to paroxetine (20 mg/d) or mirtazapine (30 mg/d) for a period of 6 weeks; half the dosage was given in the first week. During the preceding 1-month baseline and 6-week treatment period, intravaginal ejaculation latency times were measured at home using a stopwatch procedure. The trial was completed by 18 men. Analysis of variance revealed a between-group difference in the development of the delay in intravaginal ejaculation latency time over time (P < 0.001); the intravaginal ejaculation latency time after paroxetine and mirtazapine gradually increased from 15 to 119 s and from 23 to 28 s, respectively, after 6 weeks. Paroxetine 20 mg/d exerted a strong delay (maximum 5.7-fold increase), whereas mirtazapine 30 mg/d did not delay ejaculation (0.9-fold increase). These results confirm earlier findings that paroxetine, but not mirtazapine, significantly delays orgasm and ejaculation in men with early ejaculation, whereas mirtazapine is devoid of any effect on it. PMID- 14520125 TI - Hyperprolactinemia in patients on antipsychotic drugs causes ADP-stimulated platelet activation that might explain the increased risk for venous thromboembolism: pilot study. AB - Recently, an increased risk of venous thromboembolism in patients on antipsychotic drugs has been reported, but the molecular etiology is still unknown. Most antipsychotic drugs act as dopamine antagonists, and some of them cause hyperprolactinemia. Hyperprolactinemia has recently been found to cause increased platelet activation via potentiating ADP effects on human platelets. We assessed prolactin values as well as ADP-stimulated and thrombin receptor activator 6-stimulated expression of the platelet activation marker P-selectin in 20 consecutive patients under therapy with antipsychotic drugs. We detected a significant correlation between prolactin values and ADP-stimulated P-selectin expression on platelets in patients on antipsychotic drugs, revealing a significant higher platelet stimulation in hyperprolactinemic patients on antipsychotic drugs than in normoprolactinemic controls. Therefore, hyperprolactinemia might be the yet unknown acquired risk factor in patients on antipsychotic drugs explaining the increased risk for venous thromboembolism in these patients. PMID- 14520126 TI - Potential mechanisms of action of lamotrigine in the treatment of bipolar disorders. AB - Based on the mood-stabilizing properties of carbamazepine and valproate, new anticonvulsants have been explored for use in bipolar disorders. One such agent, lamotrigine, has a novel clinical profile in that it may "stabilize mood from below," as it appears to maximally impact depressive symptoms in bipolar disorders. In this paper, we review the mechanisms of action of lamotrigine in an effort to understand the basis of its distinctive clinical use in the management of bipolar disorders as well as its diverse antiseizure effects. We consider lamotrigine mechanisms, emphasizing commonalities and dissociations among actions of lamotrigine, older mood stabilizers, and other anticonvulsants. Although ion channel effects, especially sodium channel blockade, may importantly contribute to antiseizure effects, such actions may be less central to lamotrigine thymoleptic effects. Antiglutamatergic and neuroprotective actions are important candidate mechanisms for lamotrigine psychotropic effects. Lamotrigine has a variable profile in kindling and contingent tolerance experiments and does not appear to have robust gamma-aminobutyric acid or monoaminergic actions. Lamotrigine intracellular signaling effects warrant investigation. Although lamotrigine mechanisms overlap those of other mood-stabilizing anticonvulsants, important dissociations suggest candidate mechanisms, which could contribute to lamotrigine's distinctive psychotropic profile. PMID- 14520127 TI - Recurrent episodes of perceptual alteration in patients treated with antipsychotic agents. AB - Antipsychotics have been sporadically reported to induce the brief recurrent episodes characterized by hypersensitivity of the visual perception, which is known as paroxysmal perceptual alteration (PPA) mainly in Japan. PPA is characterized by hypersensitivity of perception mainly in visual modalities, which could occur in patients treated with antipsychotics. PPA is occasionally known to be accompanied by an oculogyric crisis (OGC). In this study, we examine the prevalence and the characteristics of patients with PPA. Three hundred thirty eight patients who were treated with antipsychotics and diagnosed with schizophrenia, mood disorders, or neurotic disorders (International Classification of Disease, or ICD-10) were interviewed. We compared the characteristics between subjects with and without PPA. The mean overall prevalence of PPA was 3.25%, which was higher among patients treated with high potency antipsychotics (3.91%) than in subjects treated with mid-potency or low potency drugs (1.16%). PPA occurred simultaneously with OGC at the rate of 36.4%. We also found much similarity between PPA and OGC in terms of phenomenology. We suggest that PPA could manifest itself as an undesirable effect of antipsychotics, especially those of high potency. PPA and OGC may a share common underlying mechanism. PMID- 14520128 TI - Effects of topiramate on aggressive, self-injurious, and disruptive/destructive behaviors in the intellectually disabled: an open-label retrospective study. AB - This study reviews the treatment response to the antiepileptic drug topiramate (Topamax-mean dose 202 mg/d, range 150-350 mg/d) of a group of 22 institutionalized intellectually disabled adults (8 males, 14 females, mean age 46.5 years, age range 25-70 years). These individuals were predominantly classified as having severe or profound intellectual disability and as having a mood disorder. The individuals studied were treated for aggression, self injurious behaviors, destructive/disruptive behaviors or a combination of these, and/or other challenging and maladaptive behaviors. All subjects were receiving concurrent psychotropic and/or anticonvulsant medications. Effectiveness was determined by retrospective review of summaries of quarterly multidisciplinary Neuropsychiatric Behavioral Reviews. Assignment of global severity scores and evaluation of longitudinal behavioral graphs of target symptoms occurred. Overall, statistically significant decreases in global severity scores and in the cumulative aggression and worst behavior rates occurred in the subjects, especially when the 3 months before and the 3 to 6 months after starting topiramate were compared. The overall subject group showed no significant weight changes. One subject developed delirium, 1 developed hypoglycemia, 1 developed sedation, and 2 developed constipation. The results suggest that topiramate may have a role in the treatment of challenging/maladaptive behaviors in intellectually disabled individuals. PMID- 14520129 TI - Imipramine and buspirone in patients with panic disorder who are discontinuing long-term benzodiazepine therapy. AB - Pretreatment with imipramine, buspirone, or placebo was compared in 40 patients meeting the Diagnostic and Statistical Manual of Mental Disorders, Revised Third Edition criteria for panic disorder and in patients who were discontinuing long term benzodiazepine use. The average duration of benzodiazepine use was 75 +/- 64 months, and the average benzodiazepine intake expressed as diazepam equivalents was 25.7 +/- 19 mg/d. We hypothesized that pretreatment with either imipramine or buspirone, in contrast to pretreatment with placebo, would lead to a significant decrease of symptoms of anxiety and depression before tapering benzodiazepines, thus making the taper process easier to complete. All 3 treatments (imipramine, buspirone, and placebo) caused a reduction in anxiety and depression symptoms as measured by changes in the Hamilton Anxiety Rating Scale and Hamilton Depression Rating Scale. Neither discontinuation severity nor taper-free status 12 weeks posttaper differed between the 3 treatment groups. PMID- 14520130 TI - Does nefazodone improve both depression and Parkinson disease? A pilot randomized trial. AB - Some of the selective serotonin reuptake inhibitors (SSRI)-induced motor side effects are mediated by stimulating 5-HT2 receptors in the basal ganglia, probably because serotonin inhibits the subsequent neuronal dopamine release. We hypothesized that nefazodone, a serotonin 2 antagonist/reuptake inhibitor (SARI) that selectively blocks 5-HT2 receptors, could disrupt the aforementioned inhibitory pathway. Therefore, increased dopamine levels in the postsynaptic milieu and an improvement in the motor symptoms in depressed patients with Parkinson disease (PD) should be observed. This study was designed to determine whether nefazodone has a dual activity as an antidepressant and as an agent capable of reducing the extrapyramidal symptoms in depressed parkinsonian patients. Depressed patients with PD were randomly assigned to 2 therapeutic groups: nefazodone or fluoxetine. Patients were evaluated by a psychiatrist and were blindly assessed by a neurologist with an array of scales. Patients on nefazodone (n = 9) showed a significant improvement over time in the total Unified Parkinson Disease Rating Scale score (UPDRS) (part II + part III) (P = 0.004) and in the UPDRS subscore part III (P = 0.003). None of these scores changed over time in the fluoxetine group (n = 7). Both, nefazodone and fluoxetine were equally effective as antidepressants: Beck Depression Inventory scores significantly improved (P < 0.001), with no significant differences between treatment groups (P = 0.97). If our results can be confirmed in a larger clinical trial, nefazodone ought to be considered over fluoxetine given its secondary beneficial effects regarding the reduction of extrapyramidal symptoms in depressed PD patients. PMID- 14520132 TI - Fluoxetine augmentation of haloperidol in chronic schizophrenia. PMID- 14520131 TI - Treatment of acute alcohol withdrawal with gabapentin: results from a controlled two-center trial. AB - A few case reports and data from animal experiments point to a possible efficacy of gabapentin (GP) in the treatment of alcohol withdrawal syndrome (AWS). Because of ethical considerations, the efficacy of GP in acute AWS was tested in an add on fashion to clomethiazole (CLO). Given that the symptom-triggered amount of CLO required to limit AWS within the first 24 hours is related to the severity of AWS, we tested this amount of CLO during placebo (P) or GP (400 mg qid) under double blind, randomized conditions. Sixty-one patients (P = 29/GP = 32) suffering from alcohol dependence (ICD-10) and without any other psychiatric condition or psychotropic medication were included. The groups were not significantly different in baseline characteristics (eg, demographic data, severity of AWS). Both ITT and completer analyses revealed no significant differences between the groups considering the primary effectiveness measure: amount of CLO required in the first 24 hours (P = 6.1 +/- 5.4/GP = 6.2 +/- 4.7 capsules). In addition, premature discontinuations (P = 3/GP = 2) and decreases in Mainz Alcohol Withdrawal Scores were not significantly different in the first 48 hours of AWS (secondary effectiveness measures). Tolerability of combined CLO/GP was studied throughout the whole treatment comprising a 5-day lasting reduction part subsequent to the first 48 hours. Throughout the whole 7-day treatment a total of 5 and 2 patients dropped out and 6 and 5 patients reported adverse clinical events in the P and GP groups, respectively. All together, GP (400 mg qid) was no better than P in saving initial consumption of CLO or decreasing initial Mainz Alcohol Withdrawal Scores suggesting that GP was ineffective in the management of acute AWS in this model. The combination of GP and CLO was safe. PMID- 14520133 TI - Donepezil for the treatment of a schizophrenia patient with frontal lobotomy. PMID- 14520134 TI - Late-onset agranulocytosis in a patient with schizophrenia after 17 months of clozapine treatment. PMID- 14520135 TI - The effect of olanzapine on anxiety among patients with schizophrenia: preliminary findings. PMID- 14520136 TI - Susceptibility to episode recurrence in spontaneous recurrence of methamphetamine psychosis. PMID- 14520138 TI - St. John's wort in generalized anxiety disorder: three more case reports. PMID- 14520137 TI - Abnormal CSF monoamine metabolism in serotonin syndrome. PMID- 14520139 TI - Use of linezolid in children. Introduction. PMID- 14520140 TI - Linezolid pharmacokinetics in pediatric patients: an overview. AB - BACKGROUND: There are a number of physiologic and developmental differences between children and adults that can influence the absorption, distribution, metabolism and elimination of a drug. Therefore it is important to determine the specific pharmacokinetic characteristics for individual drugs in pediatric patients so that appropriate age-specific dosage regimens can be developed and evaluated in clinical trials. This review summarizes the pharmacokinetic parameters of linezolid in pediatric patients and the rationale for the approved dosing recommendations for this population. METHODS: The pharmacokinetics of linezolid in pediatric patients has been evaluated in 4 clinical trials, including >180 patients ranging in age from preterm newborn infants up to 18 years of age. In all of these studies, patients received a single intravenous dose of linezolid. Plasma linezolid concentrations have been determined by validated high performance liquid chromatography (adult studies) or liquid chromatography/mass spectrometry/mass spectrometry (pediatric studies) methods. RESULTS: The pharmacokinetics of linezolid, especially elimination clearance, is age-dependent. Children younger than 12 years of age have a smaller area under the drug concentration-time curve, a faster clearance and a shorter elimination half-life than adults. Although clearance rates in newborn infants are similar to those in adults, clearance increases rapidly during the first week of life, becoming 2- to 3-fold higher than in adults by the seventh day of life. The clearance of linezolid decreases gradually among young children, becoming similar to adult values by adolescence. The pharmacokinetics of linezolid in children age 12 years and older is not significantly different from that of adults. CONCLUSIONS: Because of the higher clearance and lower area under the drug concentration-time curve, a shorter dosing interval for linezolid is required for children younger than 12 years of age to produce adequate drug exposure against target Gram-positive pathogens. PMID- 14520141 TI - Linezolid versus vancomycin in the treatment of known or suspected resistant gram positive infections in neonates. AB - BACKGROUND: Gram-positive infections caused by susceptible and resistant strains of Staphylococcus aureus, coagulase-negative staphylococci and enterococci are increasing problems in neonates. Linezolid, a new oxazolidinone, is active against these pathogens and has recently been approved by the Food and Drug Administration for treating Gram-positive infections in pediatric patients. OBJECTIVE: To compare the clinical efficacy and safety of intravenous and oral linezolid with vancomycin (10 to 15 mg/kg every 6 to 24 h) in neonates (age 0 to 90 days). METHODS: Hospitalized infants with known or suspected hospital-acquired pneumonia, complicated skin or skin structure infections, bacteremia or other infections (e.g. pyelonephritis, abdominal abscess) were eligible. Test-of-cure clinical response was evaluated at follow-up. RESULTS: Sixty-three neonates, randomized 2:1 to linezolid (n = 43) or vancomycin (n = 20) were included in the intent-to-treat group. Clinical cure rates at follow-up in the intent-to-treat group were higher, but not significantly different, for linezolid vs. vancomycin (78% vs. 61%; P = 0.196). Corresponding cure rates in clinically evaluable patients were 84% vs. 77% (P = 0.553) for linezolid and vancomycin, respectively. Pathogen eradication rates were as follows in the linezolid and vancomycin groups, respectively: S. aureus (67% vs. 60%; P = 0.850); coagulase-negative staphylococci (88% vs. 100%; P = 0.379); and enterococci (71% vs. 0%; P = 0.168). Results for hematology and chemistry assays were similar between treatment groups. Fewer linezolid-treated neonates had drug-related adverse events than vancomycin-treated neonates (12% vs. 32%; P = 0.058). CONCLUSIONS: Linezolid is well-tolerated and as effective as vancomycin in the treatment of resistant Gram positive infections in neonates. PMID- 14520142 TI - Linezolid for the treatment of children with bacteremia or nosocomial pneumonia caused by resistant gram-positive bacterial pathogens. AB - BACKGROUND: Nosocomial infections, particularly hospital-acquired pneumonia (HAP) and bacteremia, are an increasing concern in pediatric hospitals and pediatric intensive care units. Gram-positive pathogens are a leading cause of these infections in children. Linezolid is well-tolerated and as effective as vancomycin in the treatment of these infections in adults. OBJECTIVE: To evaluate the clinical effectiveness and safety of iv/oral linezolid and iv vancomycin in children with resistant Gram-positive HAP or bacteremia. METHODS: Hospitalized children <12 years of age were randomized 2:1 to linezolid or vancomycin. Patients received linezolid 10 mg/kg iv every 8 h with the option to change treatment to oral linezolid suspension 10 mg/kg every 8 h or iv vancomycin 10 to 15 mg/kg every 6 to 24 h. Clinical response was evaluated at follow-up. Results from an analysis of patients with HAP or bacteremia are presented. RESULTS: Thirty-nine patients (linezolid, 23; vancomycin, 16) with HAP and 113 patients with bacteremia (linezolid, 81; vancomycin, 32) were included in the intent-to treat group. Clinical cure rates for clinically evaluable patients with HAP did not differ between treatment groups (linezolid, 90.0% and vancomycin, 100%; P = 0.305). No significant difference was seen in clinical cure rates in the clinically evaluable population between the linezolid and vancomycin groups for patients with catheter-related bacteremia (84.8 and 80.0%, respectively; P = 0.716) or patients with bacteremia of unknown source (79.2 and 69.2%, respectively; P = 0.501). In this subset fewer linezolid-treated patients had drug-related adverse events than did vancomycin-treated patients (19.4% vs. 28.3%; P = 0.230). Similar percentages of patients with laboratory abnormalities, including selected hematologic parameters, were seen in both treatment groups. CONCLUSIONS: Intravenous/oral linezolid was well-tolerated and as effective as vancomycin in treating children with resistant Gram-positive HAP or bacteremia. PMID- 14520143 TI - Linezolid for the treatment of complicated skin and skin structure infections in children. AB - BACKGROUND: Gram-positive pathogens are a major cause of complicated skin and skin structure infections (CSSSIs) in children. Many pathogens are developing decreased susceptibility to currently used antibiotics, increasing the need for new therapies. Linezolid is well-tolerated and effective in the treatment of these infections in adults. OBJECTIVE: To evaluate the clinical efficacy and safety of iv/oral linezolid and iv vancomycin in children with Gram-positive CSSSIs. METHODS: Hospitalized children <12 years of age were randomized (2:1 ratio) to receive either linezolid 10 mg/kg iv every 8 h (with the option to change treatment to oral linezolid suspension 10 mg/kg every 8 h) or iv vancomycin 10 to 15 mg/kg every 6 to 24 h (according to age). Clinical response, tolerance and safety were evaluated at follow-up. The results of a subset analysis of patients with CSSSIs are presented here. RESULTS: One hundred twenty intent-to-treat patients (linezolid 80, vancomycin 40) with CSSSI were included in this analysis. Clinical cure rates for clinically evaluable patients with CSSSI did not differ between treatment groups (linezolid, 93.2% vs. vancomycin, 90.0%; P = 0.594). Significantly fewer linezolid-treated patients experienced drug-related adverse events than did vancomycin-treated patients (23% vs. 48%; P = 0.006). The percentages of patients with laboratory abnormalities, including selected hematologic parameters, were generally low and similar between the treatment groups. CONCLUSIONS: Linezolid given iv or orally was well-tolerated and safe. It was as effective as vancomycin in treating children with Gram positive CSSSIs. PMID- 14520144 TI - Linezolid for the treatment of methicillin-resistant Staphylococcus aureus infections in children. AB - BACKGROUND: Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) are becoming increasingly prevalent. Linezolid is effective and well tolerated in the treatment of adults with MRSA infections. OBJECTIVE: To evaluate the clinical efficacy and safety of iv/oral linezolid in children with MRSA infections. METHODS: Data were obtained from two independent clinical trials. In an outpatient trial children (5 to 17 years of age) with uncomplicated skin and skin structure infections (SSSIs) were treated with linezolid or cefadroxil. In an inpatient trial hospitalized children (0 to 11 years of age) with pneumonia, bacteremia or complicated SSSI caused by resistant Gram-positive pathogens were administered iv linezolid with the option to switch to oral suspension (patients >90 days of age) or iv vancomycin. A subset of patients with MRSA infections from the two clinical trials is analyzed herein. RESULTS: In the outpatient trial children with skin infections caused by MRSA were treated with linezolid (15 patients) and cefadroxil (10 patients). In the microbiologically evaluable population, the clinical cure rate was 92.3% in the linezolid group and 85.7% in the cefadroxil group (P = 0.64). The pathogen eradication rate for MRSA was 92.3 and 85.7% in the linezolid and cefadroxil groups, respectively (P = 0.64). There were very few adverse events or drug-related adverse events and no serious adverse events in the outpatient trial. In the inpatient trial 20 children treated with linezolid and 14 treated with vancomycin had infections caused by MRSA. In the microbiologically evaluable population, the clinical cure rate was 94.1% in the linezolid group and 90.0% in the vancomycin group (P = 0.69). Pathogen eradication rates were 88.2 and 90.0% for the linezolid and vancomycin groups, respectively (P = 0.89). Susceptibility patterns of the MRSA isolates showed distinct patterns between the outpatient and inpatient trials. In the inpatient trial fewer patients in the linezolid group had drug-related adverse events than did those in the vancomycin group (20% vs. 43%; P = 0.15). CONCLUSIONS: Intravenous/oral linezolid is effective and well-tolerated in children with MRSA infections. PMID- 14520145 TI - Hematologic effects of linezolid in young children. AB - BACKGROUND: Linezolid is an effective and well-tolerated antibiotic for the treatment of Gram-positive infections, including hospital and community-acquired pneumonia and complicated and uncomplicated skin and skin structure infections. In adults linezolid treatment for >/=2 weeks has been associated with reversible hematopoietic suppression, primarily thrombocytopenia. OBJECTIVE: To evaluate the occurrence of hematologic effects in children with Gram-positive infections in an open label study of linezolid vs. vancomycin. METHODS: Detailed analyses of hematologic data, including reported hematologic adverse events, complete blood counts, reticulocyte index (RI) and iron studies (serum iron and transferrin saturation), were conducted in both groups at baseline and during and after treatment with the use of an intent-to-treat analysis. RESULTS: Three hundred sixteen patients (median age, 1.65 yr) randomized 2:1 to linezolid (n = 215) or vancomycin (n = 101) were treated. Total treatment durations were similar in the vancomycin group (12.2 +/- 6.4 days; median, 11.0 days) and the linezolid group (11.3 +/- 5.0 days; median, 11.0 days) (P = 0.20). No significant differences were noted in drug-related hematologic events, such as thrombocytopenia (linezolid, 1.9% vs. vancomycin, 0%; P = 0.170), anemia (linezolid, 1.4% vs. vancomycin, 1.0%; P = 0.771) or neutropenia (linezolid, 0% vs. vancomycin, 0%). Hemoglobin values also were similar between treatment groups when assessed by shifts from baseline to lowest recorded value. Frequency of occurrence of any substantially abnormal value for hemoglobin (15.7% vs. 12.4%), platelets (12.9% vs. 13.4%) and neutrophils (5.9% vs. 4.3%) were similar in the linezolid and vancomycin groups. No clinically relevant changes in RI or iron studies were noted between treatment groups, and parallel increases in RI occurred with both linezolid and vancomycin. CONCLUSIONS: No significant differences in hematologic profiles between linezolid and vancomycin occurred in this pediatric population. PMID- 14520147 TI - Compliance barriers in glaucoma: a systematic classification. AB - PURPOSE: To systematically identify and describe common obstacles to medication adherence (i.e., compliance) for patients with glaucoma. METHODS: A prospective case series of structured interviews were conducted with 48 patients with glaucoma. The subjects' responses were recorded verbatim on interview forms as well as recorded on audiotapes. Situational obstacles to medication adherence were elicited. Using hierarchical cluster analysis, the situational descriptions were stratified, grouped, and analyzed by frequency distribution. RESULTS: Seventy-one unique situational obstacles were reported. These were then grouped into 4 defined and separate categories: situational/environmental factors (35 of 71 situations; 49%), medication regimen (23 of 71; 32%), patient factors (11 of 71; 16%), and provider factors (2 of 71; 3%). CONCLUSION: Significant barriers to compliance exist for patients with glaucoma in addition to those cited by previous ophthalmic studies. A systematic classification (i.e., taxonomy) of these barriers was formulated to assist in optimizing patient education and problem-solving regarding prescribed therapeutic regimens. PMID- 14520146 TI - Safety and tolerability of linezolid in children. AB - BACKGROUND: Linezolid, an oxazolidinone, is effective in the treatment of adults and children with community-acquired and nosocomial pneumonia and uncomplicated and complicated skin and skin structure infections (SSSIs), including infections caused by Gram-positive resistant pathogens. Because of the increasing use of linezolid, it is important to review the common adverse events (AEs) associated with its use in children with the use of data from clinical trials. OBJECTIVE: The safety and tolerability of linezolid in pediatric patients with Gram-positive infections were determined in four pediatric clinical studies. Study I included pediatric patients with community-acquired pneumonia; Study II included otitis media; Study III included SSSIs; and Study IV included complicated SSSIs, nosocomial pneumonia and bacteremia. METHODS: Studies I and II had no comparator arm. Study III was randomized and compared linezolid with cefadroxil. Study IV also was randomized and compared linezolid with vancomycin. Patients <12 years of age received linezolid 10 mg/kg; patients age 12 years and older received 600 mg (intravenous/oral). Dosing frequency (two to three times daily) varied depending on age and clinical diagnosis. The primary safety endpoints were AEs, drug related AEs, serious AEs and selected laboratory tests. RESULTS: In the 4 studies 958 patients were included in the intent-to-treat analysis. In the linezolid vs. cefadroxil study (Study III), the most common AEs in patients treated with linezolid were diarrhea (7.8%), headache (6.5%) and upper respiratory tract infection (3.7%). In the linezolid vs. vancomycin study (Study IV), the most common AEs in the linezolid group were fever (14.1%), diarrhea (10.8%) and vomiting (9.4%). The most common drug-related AEs for linezolid in all 4 studies were diarrhea, vomiting, loose stools and nausea. None of these common AEs or drug-related AEs occurred more frequently in patients treated with linezolid than in those in the comparator group. CONCLUSIONS: Linezolid was safe and well tolerated in pediatric patients with community-acquired pneumonia, otitis media, SSSIs and infections caused by Gram-positive resistant pathogens. PMID- 14520148 TI - Repeat reliability of the multifocal visual evoked potential in normal and glaucomatous eyes. AB - PURPOSE: To investigate the repeat reliability of the multifocal visual evoked potential (mfVEP). PATIENTS AND METHODS: Fifteen subjects with no known abnormalities of the visual system and 10 patients with glaucoma participated in the study. Monocular mfVEPs were recorded on two separate days, using a 60 sector, pattern-reversal dart board array. Within a single session, two 7-minute. recordings were obtained for each eye. The amplitude of each mfVEP response was obtained using a root mean square measure (RMS). An mfVEP ratio [10*log (RMS day 1 / RMS day 2)] provided a measure of the reproducibility of an individual response. The same calculations were performed for Run 1 compared with Run 2 within a day and Run 1 (Run 2) compared with Run 1 (Run 2) across days. RESULTS: For all 1800 mfVEP responses (60 sectors x 15 subjects x 2 eyes), the correlation between the amplitude on day 2 and the amplitude on day 1 was good (r = 0.85). The mean standard deviation (SD) of the 60 mfVEP ratios for the individual subjects was 1.63 dB for the 14-minute records (the combination of the two 7 minute recordings). On average for the 7-minute records, the mean SD across days was 1.77 dB while the mean SD within a day was 1.53 dB. The correlation within a day (r = 0.87) also was slightly larger than across days (r = 0.80). The mean SD decreased as the RMS amplitude increased. The patients' mean SD was 1.75 dB with r equal to 0.82. CONCLUSIONS: The repeat reliability of the mfVEP was good (approximately 1.6dB); in fact, it was better than that typically obtained with static automated perimetry (approximately 2.7dB). Repeat testing on separate days added surprisingly little to the variability seen with repeat testing within the same session. PMID- 14520150 TI - Staging of functional damage in glaucoma using frequency doubling technology. AB - PURPOSE: To evaluate three new approaches for staging severity of glaucomatous visual field defects using frequency doubling technology. PATIENTS AND METHODS: One hundred and four patients with either ocular hypertension or chronic glaucoma were tested with both standard automated perimetry (SAP, 30-2 Humphrey threshold test) and frequency doubling technology (N-30 threshold test). Standard automated perimetry results were classified into four groups (normal tests, early defects, moderate defects, and severe defects) using the Glaucoma Staging System. Frequency doubling technology tests were also classified in four groups using three different approaches: frequency doubling technology probability map analysis, considering the number and location of disturbed points, frequency doubling technology MD and PSD indices, graphed on a two-axis diagram (FDT Staging System), and an abnormality score, based on both the statistical significance and the spatial location of depressed points. A control group of 20 eyes from 20 normal subjects was also tested and classified in the same way. The Cohen Kappa was used to compare the level of agreement between the three frequency doubling technology methods of classification and the glaucoma staging system. RESULTS: Measure of agreement was 0.679 using probability map assessment, 0.793 using the frequency doubling technology staging system, and 0.663 using the abnormality score. The specificity rate was 95% for all three methods. CONCLUSIONS: All the studied approaches were able to correctly stage the glaucomatous functional damage, but the frequency doubling technology staging system was the easiest and quickest method. Moreover, it is the only method that supplies information on the characteristics of the defect, without requiring any other time-consuming procedures. PMID- 14520149 TI - Rate of optic disc cup progression in treated primary open-angle glaucoma. AB - PURPOSE: To quantitatively evaluate the rate of cup-to-disc ratio progression in treated patients with primary open-angle glaucoma and to identify clinical factors associated with cup progression. PATIENTS AND METHODS: Fifty one eyes of 51 treated primary open-angle glaucoma patients with a minimum of 9-year longitudinal series of stereoscopic optic disc photographs were studied. Eyes with any other ocular disease except for mild cataract were excluded. Each set of stereoscopic photographs was digitized and viewed stereoscopically on a computer screen using a hand-held stereoscope. Computer-aided planimetry was performed on each set of photographs with examiner-defined cup and disc margins using custom made software. The software computed linear cup-to-disc ratios as well as peripapillary atrophy area. Both inter-observer and intra-observer reliabilities were evaluated in a masked, random fashion using intra-class correlation. Changes in linear cup-to-disc ratios and peripapillary atrophy were estimated using linear regression over time. All available clinical factors were evaluated for association with the rate of cup progression using a multiple regression model. RESULTS: All patients studied were Caucasian; 31(61%) were females. The mean age at the beginning of the study was 61.6 +/- 7.8 years (range 46-81). The mean follow-up period was 14.3 +/- 3.5 years (median 14.0, range 9.6-22.3). A total of 173 sets of stereo disc photographs were analyzed (3.4 +/- 1.3 per patient). The initial and final linear cup-to-disc ratios were 0.64 +/- 0.15 and 0.73 +/- 0.14 respectively. The inter-rater and intra-rater intraclass correlations were 0.76 (95% CI 0.61-0.87) and 0.97 (95% CI 0.93-0.98) respectively. Using linear regression the rate of linear cup-to-disc ratio change was 0.0068 +/- 0.0062 per year (range -0.0025- 0.0269). Three eyes had an increase in the peripapillary atrophy area. The higher yearly average intraocular pressure was significantly associated with faster rate of linear cup-to-disc ratios progression (P = 0.03). CONCLUSIONS: In treated patients with primary open-angle glaucoma, the rate of progressive optic disc cupping was slow (LCDR progression of 0.0068 per year). The higher yearly average intraocular pressure was significantly associated with a faster rate of cup progression. PMID- 14520151 TI - Bleeding during gonioscopy after deep sclerectomy. AB - PURPOSE: To show a new complication after deep sclerectomy (DS). METHODS: We described two eyes of two patients with open-angle glaucoma and cataract who were operated on of an uneventful phacoemulsification and DS with SK-gel implantation. RESULTS: Bleeding during gonioscopic examination occurred in both eyes 7 and 8 months after combined surgery. The blood originated from the vessels around the Descemet window, and was probably due to manipulation or rocking of the goniolens. Pressure was immediately applied to the gonioscopic lens and the hyphema was interrupted. CONCLUSION: These cases show the presence of new vessels around the Descemet window after DS with SK-gel. Bleeding from the Descemet window vessels can occur during gonioscopy even months after DS. We recommend conducting a careful gonioscopic examination in patients who have undergone DS to avoid this complication. PMID- 14520152 TI - Correlation between the early morphological appearance of filtering blebs and outcome of trabeculectomy with mitomycin C. AB - PURPOSE: To correlate the morphologic appearance of filtering blebs in the early postoperative period with the outcome of trabeculectomy with mitomycin C (MMC) during the first postoperative year. PATIENTS AND METHODS: In a prospective study, the morphologic appearance of filtering blebs after primary trabeculectomy with adjunctive MMC (0.1 mg/ml for 5 minutes intra-operatively) was classified; 49 eyes of 49 patients were examined preoperatively, 1 and 3 days, 1 and 2 weeks, 1, 3, 6, and 12 months postoperatively. Status of filtering bleb, intraocular pressure (IOP), and number of medications were recorded. RESULTS: One year after surgery all patients had IOP < or = 21; 6 patients received antiglaucoma medication. One eye required needling of the filtering bleb because of encapsulation. During the first postoperative year, eyes with conjunctival subepithelial micro cysts, observed in the first and the second postoperative week, had significantly lower mean IOP, than eyes without (11.1 mm Hg vs. 13.9 mm Hg; p:0.0043, ANOVA). Eyes with corkscrew vessels, observed in the first and the second postoperative week, had significantly higher mean IOP, than eyes without during the first postoperative year (13.4 mm Hg vs. 11.7 mm Hg; p:0.0141, ANOVA). CONCLUSION: Classification of filtering blebs after trabeculectomy with MMC may help to disclose patients with an increased failure risk. PMID- 14520153 TI - Influence of serum levels of sex hormones on intraocular pressure in menopausal women. AB - PURPOSE: The purpose of this study was to assess the effect that changes in sex hormone levels have on intraocular pressure (IOP) in menopausal women. SUBJECTS AND METHODS: Thirty menopausal women on hormone replacement therapy and 32 menopausal women who had never received hormone replacement therapy (HRT), matched for age and duration of amenorrhea, participated in this study. Intraocular pressures were measured with Goldmann applanation tonometer. Serum levels were measured for estradiol, free testosterone, and follicle stimulating hormone (FSH). The influence of serum hormone levels on IOP was assessed by correlation analysis. RESULTS: The mean IOP of postmenopausal women receiving HRT (13.29 +/- 2.28 mm Hg) was not significantly different from that of menopausal women not receiving HRT (13.56 +/- 2.5 mm Hg, P = 0.24). Higher testosterone levels were associated with higher IOPs in women receiving HRT (r = 0.48, P = 0.02) and in those not receiving HRT (r =0.42, P = 0.003). No significant correlations were observed between IOP and serum levels of estradiol and FSH in either group. CONCLUSION: Our data provide evidence for a relation between serum testosterone levels and IOP in menopause. Higher testosterone seems to have a tendency to increase IOP in menopausal women. PMID- 14520154 TI - Failed non-penetrating surgery. PMID- 14520155 TI - [Possible mechanisms responsible for elevated IOP in POAG patients]. PMID- 14520156 TI - Thin corneas and the risk of progression. PMID- 14520157 TI - Malignant glaucoma, an enigma solved? PMID- 14520158 TI - Neuroprotection in Huntington's [correction of Hungtington's] disease. PMID- 14520159 TI - Improvement of ataxia in cortical cerebellar atrophy with the drug gabapentin. PMID- 14520160 TI - Poor tolerability of a transdermal nicotine treatment in Parkinson's disease. AB - Studies assessing the effect of transdermal nicotine in Parkinson's disease (PD) have generated mixed results regarding its efficacy to treat motor and cognitive deficits. These studies generally reported good tolerability in nonsmoking PD patients. The authors report the tolerability data of an open trial with transdermal nicotine in PD. Twenty-two therapeutically well-controlled nonsmoking PD patients received a transdermal nicotine treatment over 25 days according to the following fixed titration schedule: 7 mg for the first 11 days, 14 mg for the next 11 days, and 21 mg for the last 3 days. Fourteen PD patients (64%) had side effects such as nausea, vomiting, and dizziness, and 10 of them withdrew from the study. Factors such as age, body mass index, disease duration, and motor disability were not related to this intolerance. Transdermal nicotine can produce unpleasant adverse effects in patients with PD. Given that similar doses of nicotine were better tolerated in previous studies, the authors suspect the pharmacokinetic profile of the transdermal delivery system to be a determining factor in the effect of nicotine treatment in PD. PMID- 14520161 TI - Addition of a dopamine agonist, cabergoline, to a serotonin-noradrenalin reuptake inhibitor, milnacipran as a therapeutic option in the treatment of refractory depression: two case reports. AB - We illustrate 2 patients with depression who attained dramatic improvement of energy loss and fatigue when treated with cabergoline, a dopamine agonist, and milnacipran, a serotonin-noradrenalin reuptake inhibitor. Although the biologic basis of energy, motivation, and fatigue in association with depression remains unknown, some reports suggest that the decrease of noradrenalin and dopamine in the brain are particularly related to these symptoms. Therefore, treatment strategy that enhances these two monoamine neurotransmissions may be appropriate for getting a boost in energy and eliminating fatigue in patients with depression. These cases suggest that further studies are warranted to confirm the potential benefit of this strategy in the treatment of patients with depression who failed to attain complete remission due to residual symptoms including energy loss and fatigue refractory to previous treatments. PMID- 14520162 TI - Lamotrigine treatment for post-stroke pathological laughing and crying. AB - Pathologic laughing and crying (PLC) is a common distressing and socially disabling condition in stroke patients. Antidepressants, particularly selective serotonin reuptake inhibitors (SSRIs), have been increasingly recognized as the treatment of choice for pathologic crying (PC). However, little is known about etiologies and other treatment options for various clinical manifestations of PLC. This case report illustrates the beneficial effect of lamotrigine, a novel antiepileptic drug with antidepressant and mood-stabilizing properties in post stroke PLC. A 60-year-old woman developed PLC after an ischemic stroke affecting the left frontal and temporal lobes. She was treated with lamotrigine initially at the dose of 50 mg a day, which was gradually increased to 100 mg a day over a 4-week period. There was a significant and rapid recovery in both laughing and crying components of PCL with lamotrigine treatment. The symptoms of pathologic laughing have shown a better response to lamotrigine than PC. Controlled investigations are needed to evaluate the beneficial as well as the differential effects of lamotrigine on PLC. PMID- 14520163 TI - Drug-induced pseudotumor cerebri. AB - Pseudotumor Cerebri (PTC) is an uncommon disorder whose etiology is largely unknown, although its association with steroid withdrawal, hypervitaminosis A, and the use of the tetracycline group of drugs has been well documented. We report here a case in which a patient on chronic divalproex therapy for a seizure disorder developed PTC. Changing his antiepileptic medication from divalproate to topiramate effected a remission of PTC symptoms while maintaining his seizure free status. It is recommended that physicians treating epilepsy, vascular headaches, or mood disorders with divalproate consider the diagnosis of PTC when their patients complain of new onset of headaches or an increase in frequency or severity of existing headaches--especially those associated with a visual disturbance--to prevent permanent visual loss. PMID- 14520164 TI - The effects of a cholinesterase inhibitor are prominent in patients with fluctuating cognition: a part 3 study of the main mechanism of cholinesterase inhibitors in dementia. AB - Fluctuating cognition is evidenced in different forms of dementia and is accompanied by electroencephalographic (EEG) abnormalities. The authors hypothesize that cholinesterase inhibitors are effective mostly in patients with fluctuating cognition. Twenty-three patients affected by mild dementia with similar scores on Mini-Mental State Examination (MMSE), Alzheimer Disease Assessment Scale-cognitive subscale (ADAS-cog), and Unified Parkinson's Disease Rating Scale evaluation were classified in a group with fluctuating cognition (n = 11) and a group of nonfluctuators (n = 12). All patients were assigned randomly to the branches of a double-blind crossover study of donepezil (DPZ), a 5 to 10 mg dose, versus vitamin E, a 2000 IU dose, for 30 days. MMSE, ADAS-cog, University of California at Los Angeles Neuropsychiatric Inventory (NPI), quantitative EEG, P3 event-related potentials, choice reaction time variability (CRTV) were assessed at baseline and at the end of treatments. At the end of the crossover study all patients received DPZ for 6 months. The dominant EEG frequency variability, low EEG frequencies amplitude, the P3 latency and jitter, CRTV, and NPI was significantly different in the fluctuating cognition group than the nonfluctuating group at baseline (P < 0.001). Short-term DPZ administration induced a significant increase in MMSE scores, reduction of ADAS-cog and of NPI scores (P < 0.003-0.001), increase of EEG alpha activity and reductions of P3 latency and jitter, dominant frequency variability and CRTV (P < 0.009-0.001) in the fluctuating cognition group, and significant increases of MMSE scores (P = 0.03) and a decrease of P3 jitter and dominant frequency variability (P < 0.034 0.041) in the nonfluctuating group. Short-term DPZ effects differed significantly between fluctuating cognition and nonfluctuating patients (0.001). Significant effects of the 6-month observation were observed only in fluctuating cognition patients. Logistic analysis showed that P3 latency predicts the effect of DPZ (P = 0.04, P < 0.01) in the crossover study, and CRTV predicts the effect at the 6 month follow-up. PMID- 14520165 TI - Dosing regimen effects of modafinil for improving daytime wakefulness in patients with narcolepsy. AB - In a multicenter, randomized, double-blind study the authors compared the efficacy of modafinil 400 mg once daily, 400 mg given in a split dose, or 200 mg once daily for maintaining wakefulness throughout the day in patients (N = 32) with narcolepsy reporting a positive daytime response to modafinil but late afternoon/evening sleepiness. Efficacy evaluations included an extended Maintenance of Wakefulness Test (9:00 am to 9:00 pm), the Clinical Global Impression of Change scale, and the Epworth Sleepiness Scale. Modafinil demonstrated significant improvement in wakefulness as assessed by the Epworth Sleepiness Scale compared with placebo at baseline (all P < 0.001). Modafinil significantly improved patients' ability to sustain wakefulness, as demonstrated by mean sleep latency at week 3 compared with placebo at baseline (all P < 0.001). The 400-mg split-dose regimen improved wakefulness significantly in the evening compared with the 200-mg and 400-mg once-daily regimen (both P < 0.05). The percentage of patients rated as "much improved" or "very much improved" with respect to evening sleepiness was 27%, 82%, and 80% in the 200-mg, 400-mg once daily, and 400-mg split-dose groups, respectively. Adverse events were mild to moderate in nature and included headache, nausea, nervousness, dyspepsia, pain, and vomiting (all 6%). Some patients may benefit from 400-mg doses of modafinil taken once daily compared with 200-mg doses. A split-dose 400-mg regimen may be superior to once-daily dosing for sustaining wakefulness throughout the entire waking day. PMID- 14520166 TI - Suppression of alcohol delirium tremens by baclofen administration: a case report. AB - Delirium tremens (DT) is a clinical condition that appears in some patients affected by severe alcohol withdrawal syndrome (AWS). DT represents a serious complication, being characterized by elevated morbidity and mortality. Benzodiazepines are presently the drug of choice; however their use is related to several side effects. Baclofen is a stereoselective gamma-aminobutyric acid (GABAB) receptor agonist. Recent studies show that baclofen is able to suppress alcohol withdrawal symptoms. At present there are no data on the effects of baclofen administration in AWS complicated by DT. Here, we report a case of DT successfully treated with baclofen. This result indicates that the efficacy of baclofen in the treatment of DT should be examined in future clinical trials. PMID- 14520167 TI - Neuroprotective effect of deprenyl in sensory neurons of axotomized dorsal root ganglion. AB - Spinal motoneuron neuroprotection by deprenyl was previously reported; the present study was carried out to evaluate neuroprotectivity in the dorsal root ganglion sensory neuron. The total neuron counts were calculated, and the axotomized sensory neurons of the dorsal root ganglion were significantly lower than those of the unaxotomized sides. Three secondary and three tertiary parameters were used. The secondary parameters were: the percentages of sensory neuron increase at the axotomized side (PNIA) and at the unaxotomized side (PNIU), and the percentage of neuronal response (PNR). The tertiary parameters were: the percentages of maximal response at the axotomized side (PMRA) and at the unaxotomized side (PMRU), and the percentage of maximal relative response (PMRR). Nonlinear statistical analysis using Gaussian, quadratic and logistic models of the tertiary parameters suggested that the data were bell-shape, which indicated that the data were biphasic. The data were divided into ascending and descending sets, and linear regression. They were analyzed according to Bent hyperbola model and the ascending set was considered as a neurotrophic phase, while the descending one as a neurotoxic phase. The slops of PMRA were higher than that of PMRU, which indicates that the axotomized neurons were more sensitive than the unaxotomized neurons to the protective and neurotoxic effect of deprenyl. Moreover, the results showed that deprenyl had a proliferative effect on the dorsal root ganglion sensory neuron. PMID- 14520168 TI - Pharmacokinetics of dexamphetamine in acute stroke. AB - Pharmacokinetics of dexamphetamine was studied in 26 patients with cerebral infarct, aged 37 to 84 years. Capsules were administered orally twice daily (at 8 am and 12 am) in three doses (2.5 mg, 5 mg, and 10 mg) for 5 consecutive days (day 1 to day 5). Blood samples were collected immediately before and 1, 2, 3, 4, 5, 6, 7, 8, and 9 hours after first administration on day 1 and before, and 4 and 8 hours after administration on days 2 through 5. The dose normalized area under the plasma concentration time curve, AUC/mg/kg, was only correlated with s creatinine (P = 0.013) but not with age, sex, body mass index, neurologic prognosis, or dose (mg/kg), as established by multiple stepwise linear regression. The median terminal half life time was 14.3 hours (inter quartile range, IQR: 11.9-6.9), 13.1 (IQR: 10.8-15.9) and 14.0 (IQR: 7.4-16.4) in the 2.5 mg, 5 mg, and 10 mg groups, respectively. The median maximal plasma concentration (Cmax) was 6.6 ng/mL (IQR: 5.1-7.0), 11.6 (IQR: 7.8-12.8), and 16.9 (IQR: 14.9 20.2) in the 2.5 mg, 5 mg, and the 10 mg groups, respectively. Differences in Cmax between the 2.5 mg and 10 mg group were significant (P < 0.001). The median time to Cmax (Tmax) was 1.83 hours (IQR: 1.79-3.94), 2.59 (IQR: 1.32-3.83), and 3.86 (IQR: 1.82-5.77) in the 2.5, 5, and 10 mg groups, respectively. In the present patient population, a predetermined AUC value can be obtained by a dosing regimen of dexamphetamine based on body weight (ie, mg/kg), with precautions for patients with elevated s-creatinine. PMID- 14520169 TI - Progressive worsening of adaptive functions in Down syndrome may be mediated by the complexing of soluble Abeta peptides with the alpha 7 nicotinic acetylcholine receptor: therapeutic implications. AB - In persons with Down syndrome, soluble Abeta peptides, which result from the processing of the amyloid precursor protein, appear in the brain decades before the extracellular deposition of neuritic plaques. These soluble amyloidogenic peptides accumulate intraneuronally and can be secreted extracellularly. Their appearance has been reported in the brains of fetuses with Down syndrome. The extra gene dosage effect associated with trisomy 21 results in abnormalities of the processing of amyloid precursor protein in persons with Down syndrome. Abeta peptides, especially Abeta1-42, have been shown to form tight complexes with the alpha7 nicotinic acetylcholine receptor, interfering with transduction of the acetylcholine signal by this nicotinic receptor subtype. Furthermore, the selective binding of Abeta peptides by this nicotinic acetylcholine receptor subtype is associated with cytotoxicity. The alpha7 nicotinic acetylcholine receptor has unique electrophysiologic properties and plays a prominent role in normal psychophysiologic processes (eg, sensory inhibition) and cognition. In persons with Down syndrome there is a decrease in the ability to perform instrumental activities of daily living that worsen with aging. The progressive worsening of adaptive functions and cognition in persons with Down syndrome may be, at least in part, mediated by interference with this receptor by soluble Abeta peptides. In view of this complex formed by soluble Abeta peptides and the alpha7 nicotinic acetylcholine receptor, cholinergic interventions that have been developed for Alzheimer disease, including selective nicotinic ones, should be explored in Down syndrome. Ideally, selective cholinergic interventions would slow the progression of the worsening of adaptive function and emergence of dementia in persons with Down syndrome. PMID- 14520170 TI - Nationwide increase of Chlamydia trachomatis infection in Finland: highest rise among adolescent women and men. AB - BACKGROUND: Since 1995, the incidence of Chlamydia trachomatis infection has been increasing in Finland, although there have been no major changes in public sexually transmitted disease (STD) services or screening practice. OBJECTIVES: The objective was to study whether the change in C. trachomatis incidence is significant and to identify specific risk groups. METHODS: The incidence rates for all C. trachomatis cases notified by laboratories to the National Infectious Disease Register (NIDR) in Finland in 1995-2000 were calculated by gender, age, and domicile. Data from a sentinel STD surveillance network was used to analyze changes of risk-taking behavior in age groups with the highest C. trachomatis rates. RESULTS: During the 6-year study period, laboratory surveillance data documented an increase in the incidence rate from 23.4 per 10,000 to 29.2 per 10,000. The increase was most evident among people living in nonurban densely populated areas. Highest increase, 1.37-fold (95% confidence interval [CI], 1.29 1.46) in women and 1.69-fold (CI, 1.47-1.92) in men, occurred in the youngest age group (10-19 years old). In 2000, more women, but not men, in the age group of 10 29 years reported 5 or more annual sex partners (18.8%; CI, 16.3-21.6) than in 1995 (8.3%; CI, 5.7-11.5). CONCLUSION: National surveillance of C. trachomatis infection based on laboratory notification documented increasing incidence rates, especially among adolescents and young people. This risk group should be a target for screening and educational programs to control the epidemic of C. trachomatis infections. PMID- 14520172 TI - Spatial mixing and bridging: risk factors for what? PMID- 14520171 TI - Spatial bridges for the importation of gonorrhea and chlamydial infection. AB - A study of heterosexuals with gonorrhea and/or chlamydial infection in King County, Washington, found that 5.2% of study participants had both local and geographically distant sex partners in the 60 days before diagnosis. Individuals who served as spatial bridges were of higher socioeconomic status and older than other patients. BACKGROUND: Sexual mixing between distant geographic areas (spatial bridging) is important in the spread of antimicrobial resistance and new sexually transmitted disease pathogens. GOAL: The goal was to define the extent of sexual mixing between persons with gonorrhea or chlamydial infection in King County, Washington, and persons outside the Seattle area, and to identify characteristics of persons and partnerships associated with spatial bridging. METHODS: Patients contacted for purposes of partner notification were interviewed regarding demographics, sexual behavior, and the characteristics of their sex partners. RESULTS: Of 2912 participants, 150 (5.2%) were spatial bridgers. Bridgers were of higher socioeconomic status than nonbridgers and more often reported concurrent partnerships. Over a 39-month period, bridgers and potential bridgers linked King County with 35 states and 13 foreign countries. CONCLUSION: Spatial bridging could represent an important channel of transmission between geographic areas. These results highlight the need for linkage of prevention efforts across geographic boundaries. PMID- 14520173 TI - Non-exclusivity in adolescent girls' romantic relationships. AB - BACKGROUND AND OBJECTIVES: As adolescents develop intimate relationships skills, they might change romantic partners often. Little is known about this developmental phenomenon. GOAL: The goal was to determine the frequency of non exclusive romantic relationships in girls, the variables associated with non exclusive relationships, and the variables associated with having sexual intercourse in a non-exclusive relationship. STUDY DESIGN: Girls (12-15 years of age) participated in a longitudinal study (3 y, 7 visits) of romantic relationships. RESULTS: Seventy-two of 160 girls had a non-exclusive relationship, and of these, 23 girls had intercourse in an outside relationship. Younger age, belief that the boyfriend had an outside relationship, and less parental indirect monitoring was associated with the girl having an outside relationship. A longer time being sexually experienced and less direct parental monitoring was associated with having intercourse in the outside relationship. CONCLUSION: Non-exclusivity in adolescence is common but does not always include sexual intercourse. Healthcare providers can address non-exclusivity with adolescents directly. PMID- 14520174 TI - Development and performance of a microwell-plate-based polymerase chain reaction assay for Mycoplasma genitalium. AB - BACKGROUND: Mycoplasma genitalium is associated with, and could be the cause of, idiopathic cases of urethritis, endometritis, and cervicitis. Further epidemiologic studies on this organism are needed, but currently used polymerase chain reaction (PCR) assays are labor-intensive and culture is insensitive. GOAL: The goal was to develop and evaluate a microwell-plate-based PCR assay for M. genitalium. STUDY DESIGN: We adapted an M. genitalium PCR assay targeting the MgPa gene to a 96-microwell plate format with colorimetric detection of PCR products and incorporation of an internal inhibition control to determine the limit of detection of this assay (termed MgPa-IMW) for M. genitalium DNA and evaluate its performance on cervical and male urine specimens. RESULTS: The MgPa IMW PCR assay detected 1 and 17 genome copies of M. genitalium (with 27% and 95% confidence) and was able to detect specimens inhibited for amplification. This assay was 100% concordant (50 positive and 50 negative) with the Southern-blot based PCR assay with cervical specimens. Similarly, this test was 89% concordant with the Southern-blot-based assay for 64 male urine specimens (25 positive, 32 negative, 7 discordant), 97% concordant after correcting for specimens no longer positive by the Southern blot-based assay after freezer storage. CONCLUSION: The MgPa-IMW assay is sensitive and specific for the detection of M. genitalium in patient specimens and should facilitate large-scale screening for this organism. PMID- 14520175 TI - HIV infection risk, behaviors, and attitudes about testing: are perceptions changing? AB - BACKGROUND: People at high risk for HIV infection could be increasing their risk behaviors, especially now that improved treatments for HIV infection are available. GOAL: The goal was to investigate whether risk behaviors, perceptions of personal risk for HIV infection, and attitudes toward HIV testing among high risk persons in Oregon differed in 1996 and 1998. STUDY DESIGN: Data from the HIV Testing Survey (HITS), a cross-sectional survey administered to HIV-negative men who have sex with men (MSM), heterosexual adults at high-risk for sexually transmitted diseases (STD), and intravenous drug users (IDUs) at high risk for HIV infection in 1996 (HITS-I), were compared with data from a similar group surveyed in 1998 (HITS-II). RESULTS: Proportions of participants reporting specific risk behaviors remained relatively constant in 1996 and 1998. Personal risk of HIV infection was perceived as low by 54% of HITS-II participants and 61.2% of HITS-I participants (odds ratio [OR], 1.2; 95% confidence interval [CI], 0.9-1.7). IDUs in HITS-II were more likely than IDUs in HITS-I to perceive their risk as low (OR, 2.1; 95% CI, 1.2-3.7). CONCLUSION: Persons at high risk might underestimate their risk for HIV infection while practicing risky behaviors. The prevalence of risk behaviors in these populations could be considered the baseline against which to measure future prevention efforts. PMID- 14520176 TI - Preference among female Army recruits for use of self-administrated vaginal swabs or urine to screen for Chlamydia trachomatis genital infections. AB - BACKGROUND: Use of self-administered vaginal swabs (SAS) for the detection of Chlamydia trachomatis by nucleic acid amplification tests simplifies specimen collection and transport, especially for women in nonclinical settings. GOAL: We investigated the preference and comfort level of military women for the collection of SAS, compared with urine, for the diagnosis of genital chlamydial infections. STUDY DESIGN: During March through August 1999, female Army recruits in basic training at Fort Jackson, South Carolina, were invited to participate in the study. Participants were requested to complete a questionnaire after providing both first-void urine (FVU) and SAS specimens. Participant characteristics, preferences, and comfort levels were assessed using multivariate logistic regression. RESULTS: From 4496 eligible female recruits, 1403 (31%) completed questionnaires and 1382 provided both specimens; 11.8% (166 of 1403) of participants were infected with chlamydia. The relative sensitivity and specificity of the C. trachomatis Ligase Chain Reaction test on SAS in 1382 matched pairs was 81.1% and 98.6%, respectively, using the test result on urine specimens as the comparison standard. Most of the participants (90.8%) reported that they felt comfortable collecting the FVU specimen, and 69.6% indicated that they felt comfortable collecting SAS. Either specimen collection type received high acceptability at home and in the field, and more women reported that they would collect FVU than reported they would collect SAS in the future (in the field: FVU: 79.4%, SAS: 68.8%, P <0.001); at home: FVU: 90.9%, SAS: 82.9%, P <0.001). When questioned about ease of use, 60.4% of women reported that urine was the easier method. Preferences for SAS were associated with being white and having had sexual risk behaviors in the past 3 months. CONCLUSION: A study of preferences for urine versus self-administered vaginal swabs for the detection of C. trachomatis in military women showed that women generally found SAS acceptable. SAS should be a feasible alternative to urine collection in situations in which specimen storage or transport is an issue. PMID- 14520177 TI - Viral sexually transmitted disease vaccine acceptability among college students. AB - BACKGROUND AND OBJECTIVES: Vaccines are an important strategy for the control of infectious diseases; however, they are only successful if accepted. The object of this study was to examine factors that could affect vaccine acceptance among college students for 2 sexually transmitted diseases (STDs). GOAL: The goal was to examine the impact of gender and specific rationales on STD vaccine acceptance using health behavior theories. STUDY DESIGN: Participants completed a questionnaire regarding attitudes about hypothetical STD vaccines. RESULTS: Seventy-four percent of each group endorsed acceptance. Factors influencing genital herpes vaccine acceptance were parents' feelings, belief in vaccination, universal recommendation, numerous partners, a belief that acquisition makes finding partners difficult, and low cost. Human papillomavirus vaccine factors were parents' feelings, universal recommendation, numerous partners, safety, and low cost. CONCLUSION: Our results indicate that most college students would accept STD vaccination. Factors affecting acceptance were similar for both pathogens. The results suggest acceptance will be positively affected by health policies encouraging universal vaccination. PMID- 14520178 TI - Understanding the uneven spread of HIV within Africa: comparative study of biologic, behavioral, and contextual factors in rural populations in Tanzania and Zimbabwe. AB - BACKGROUND: Large differences in the spread of HIV have been observed within sub Saharan Africa. GOAL: The goal was to identify factors that could explain differences in the spread of HIV within sub-Saharan African populations. STUDY DESIGN: Ecologic comparison of data from population-based surveys in high and relatively low HIV prevalence rural areas in Zimbabwe, Manicaland, and Tanzania, Kisesa. RESULTS: HIV prevalence in Manicaland and Kisesa was 15.4% and 5.3% in men aged 17-44 years and 21.1% and 8.0% in women aged 15-44 years (odds ratios, 3.3 and 3.1, respectively). Marriage is later, spatial mobility more common, cohabitation with marital partners less frequent, education levels are higher, and male circumcision is less common in Manicaland. However, adjustment for differences in these factors increased the odds ratios for HIV infection in Manicaland versus Kisesa to 6.9 and 4.8 for men and women, respectively. Sexually transmitted infection levels were similar, but syphilis was only common in Kisesa. Respondents in Kisesa started sex earlier and reported more sexual partners. Age differences between partners were similar in the 2 locations. CONCLUSION: Substantial differences exist between the contemporary sociodemographic profiles of rural Manicaland and Kisesa. However, these differences did not translate into measurable differences in the biologic or behavioral factors for which data were available and did not explain the much higher HIV prevalence found in Manicaland. These findings might reflect more extensive AIDS-selective mortality and behavior change or greater bias in reporting of sexual behavior in Zimbabwe. PMID- 14520179 TI - Cervical human papillomavirus infection in the female population in Barcelona, Spain. AB - BACKGROUND AND OBJECTIVE: Human papillomavirus (HPV) is a major cause of invasive cervical cancer. Identification of HPV determinants might allow for targeting of high-risk groups for cervical cancer. GOAL: The goal was to estimate the HPV prevalence and its determinants among women from the general population of Barcelona. STUDY DESIGN: We studied a random sample of female residents in metropolitan Barcelona, Spain (n = 973). Information was obtained through personal interviews and laboratory testing of cervical exfoliated cells. HPV was detected using a GP5+/6+ polymerase chain reaction assay. RESULTS: The average age of participants was 43 years (standard deviation = 16.1 y) and the percentage of lifetime monogamy was 79%. The age-adjusted HPV prevalence was 3.0%. Independent HPV determinants were being born overseas (odds ratio [OR], 8.1; 95% confidence interval [CI], 1.9-33.5), being divorced (OR, 6.7; 95% CI, 1.9-24.3), reporting more than one sexual partner (OR, 2.6; 95% CI, 1.0-6.5), and smoking marijuana and related products (OR, 5.2; 95% CI, 1.2-21.7). Use of condoms with regular partner was protective (OR, 0.14; 95% CI, 0.02-1.0). CONCLUSION: The study confirms a low overall HPV prevalence in a largely monogamous population. The protection observed with condom use needs further evaluation. PMID- 14520180 TI - Herpes simplex virus type 1 is the prevailing cause of genital herpes in the Tel Aviv area, Israel. AB - BACKGROUND AND GOAL: The changing epidemiology of genital herpes with the increasing importance of herpes simplex virus (HSV) type 1 prompted a study on the relative prevalence of HSV-1 and HSV-2 among cases of genital herpes in the Tel Aviv area, Israel. STUDY DESIGN: A retrospective laboratory-based study of positive genital and nongenital herpes cultures performed at the Beilinson Medical Center between 1993 and 2002. Data regarding the number of isolates of each type and the age and sex of patients with genital lesions were retrieved from the database. Cultures were performed using Vero cells, and positive results were confirmed and typed by immunofluorescence. RESULTS: A total of 285 positive genital cultures and 659 positive nongenital cultures were recorded. HSV-1 was identified in 189 (66.3%) of the positive genital specimens and in 656 (99.55%) of the nongenital specimens. HSV-1 was isolated in 174 of 262 (66.4%) female subjects and 15 of 23 (65.2%) male subjects. The proportion of HSV-1 genital isolates was 72.7% in patients 15 to 24 years of age, 62% in those 25 to 44 years, and 46% in those aged 45 years or older. Overall, the annual isolation rate of genital HSV-1 has not changed markedly over the years. CONCLUSION: Herpes simplex virus type 1 has clearly been the predominant HSV type isolated from genital specimens in the Tel Aviv area over the last decade. PMID- 14520181 TI - Increasing proportion of herpes simplex virus type 1 as a cause of genital herpes infection in college students. AB - A retrospective review of genital herpes simplex virus (HSV) isolates collected in a university student health service over a 9-year period showed that an increasing proportion of isolates were HSV-1 rather than HSV-2. HSV-1 accounted for 78% of all genital isolates in this population by 2001, compared with 31% of isolates in 1993. BACKGROUND: Herpes simplex virus (HSV) type 1 is usually thought to cause less than 30% of genital herpes infections in the United States, but the proportion of infections resulting from HSV-1 is increasing in some populations. GOAL: The goal was to review the relative proportion of HSV-1 and HSV-2 as the cause of newly diagnosed genital herpes infections in a population of U.S. college students and to assess trends in the change of this proportion over time. STUDY DESIGN: Genital HSV isolates collected at a university student health service from 1993 to 2001 (n = 499) were reviewed retrospectively. Analyses included comparisons of isolates by HSV type, age group, and sex. RESULTS: The proportion of newly diagnosed genital herpes infections resulting from HSV-1 increased from 31% in 1993 to 78% in 2001 (P <0.001, linear trend P <0.001). HSV-1 was more common in females than males, but increases were noted for both sexes. HSV-1 was more common in persons aged 16 to 21 than in persons aged 22 or older. CONCLUSIONS: HSV-1 has become the most common cause of newly diagnosed genital herpes infections in this population of college students and reflects a reversal of the usual HSV-1/HSV-2 ratio. PMID- 14520182 TI - Is herpes simplex virus type 1 (HSV-1) now more common than HSV-2 in first episodes of genital herpes? PMID- 14520183 TI - Maternal-fetal surgery for spina bifida: on the brink of a new era? PMID- 14520185 TI - Informed consent, informed refusal, and informed choices. PMID- 14520184 TI - Maternal methadone dose and neonatal withdrawal. AB - OBJECTIVE: The purpose of this study was to determine whether maternal methadone dosage correlates with neonatal withdrawal in a large heroin-addicted pregnant population. STUDY DESIGN: A retrospective review of all maternal/neonatal records of pregnancies that were maintained on methadone therapy in our institution was conducted. After in-hospital stabilization, women were given daily methadone therapy under direct surveillance, with liberal dosage increases according to maternal withdrawal symptoms. Neonatal withdrawal was assessed objectively by the neonatal abstinence score. The average methadone dose in the last 12 weeks of pregnancy and the last methadone dose before delivery (cutoffs of 40, 60, or 80 mg) were correlated to various objective measures of neonatal withdrawal. RESULTS: One hundred mother/neonate pairs on methadone therapy were identified. Women who received an average methadone dose of <80 mg (n=50 women) had a trend toward a higher incidence of illicit drug abuse before delivery than women who received doses of >/=80 mg (n=50 women; 48% vs 32%; P=.1). Women who received an average methadone dose of <80 mg had similar highest neonatal abstinence score, need for neonatal treatment for withdrawal, and duration of withdrawal compared with women whose condition was maintained with dosages of >/=80 mg (score, 11.1 vs 11.5; 68% vs 66%; and 13.3 vs 13.6 days, respectively; all P>.5). For all cutoffs that were used for high versus low dose and for both the average and last methadone dosage analyses, neonatal withdrawal was similar. CONCLUSION: The maternal methadone dosage does not correlate with neonatal withdrawal; therefore, maternal benefits of effective methadone dosing are not offset by neonatal harm. PMID- 14520186 TI - Optimizing use of assisted reproduction. AB - Infertile couples who have failed to achieve pregnancy now have new options that have become available in the last two decades. The costs are generally large and frequently not covered by insurance. Risks include multiple pregnancy and hyperstimulation syndrome. Evidence concerning results of several treatment options is presented. The large difference in success between age groups is displayed. Results to be expected from donor eggs to overcome the low success in older age groups (>38 years) are presented. PMID- 14520187 TI - Reproduction in couples who are affected by human immunodeficiency virus: medical, ethical, and legal considerations. AB - There has been a transformation in the treatment of human immunodeficiency virus from the treatment of complications that define acquired immune deficiency syndrome to the maintenance of long-term health, with an expanding number of antiretroviral medications. Because human immunodeficiency virus infection now is considered to be a chronic disease, couples will be seen in greater numbers for preconception counseling. The ethical and legal implications, including the relevance of the Americans with Disability Act, are complex but support the assistance with reproduction of couples who are affected by human immunodeficiency virus in many instances. All couples who are affected by human immunodeficiency virus, whether fertile or infertile, who want to have genetically related offspring should be seen preconceptionally for counseling and testing. Intensive education involves a multidisciplinary approach to ensure that a couple is fully informed. Determination of whether to offer treatment should be based on the same criteria that are applied to couples who are affected by other chronic diseases. Medical treatment is dependent on the unique circumstances of each couple. In general, the affected partner(s) should be treated aggressively with antiretrovirals and then serum; if applicable, semen testing is required to document undetectable concentrations of human immunodeficiency virus (<50-100 copies/mL). PMID- 14520188 TI - Pregnancy rates in varying age groups after in vitro fertilization: a comparison of follitropin alfa (Gonal F) and follitropin beta (Follistim). AB - OBJECTIVE: Our purpose was to assess the efficacy of two recombinant follicle stimulating hormones, follitropin beta (Follistim, Organon, West Orange, NJ) and follitropin alfa (Gonal F, Serono, Norwell, Mass) on pregnancy rates in varying age groups of women undergoing in vitro fertilization (IVF). STUDY DESIGN: Three hundred sixty-five IVF cycles were retrospectively compared, 233 by use of follitropin beta and 132 by use of follitropin alfa, both after gonadotropin releasing hormone agonist down-regulation. Assignment to each medication was indiscriminate. The primary outcome measured was pregnancy evidenced by fetal heartbeat on ultrasonography. Secondary outcomes included days of stimulation, ampules per patient cycle, estradiol level on the day of human chorionic gonadotropin administration, total follicles present on the day of human chorionic gonadotropin administration, follicles greater than 14 mm, oocytes retrieved, mature eggs, fertilization rate, and embryos transferred. Outcomes were stratified by age, including women less than 36 years old, 36 to 39 years old, and more than 39 years old. RESULTS: There was no significant difference between follitropin beta and follitropin alfa in either the primary or secondary outcomes, although the pregnancy rate was significantly decreased with advancing age. CONCLUSION: Success rates are similar, when stratified by age, in women undergoing IVF with either follitropin beta or follitropin alfa. PMID- 14520190 TI - Clinical characteristics of patients with an abnormal clomiphene citrate challenge test. AB - OBJECTIVE: The study was undertaken to identify the clinical characteristics associated with an abnormal clomiphene citrate challenge (CC) test. STUDY DESIGN: The medical records of 123 women seen in a referral-based infertility practice who had a CC test as part of their infertility workup were reviewed. Eighty-five patients had a normal CC test and 38 patients had an abnormal test. Demographic, clinical, and laboratory characteristics were reviewed and analyzed. RESULTS: Women with abnormal CC tests were significantly older, had a lower mean body mass index, family history of early menopause, and shorter cycle intervals. Patients with an abnormal CC test had a pregnancy rate significantly lower than other women in this case series. CONCLUSION: Increased age, lower body mass index, shorter cycle intervals, lower cycle day 21 progesterone levels, and decreased pregnancy rates are associated with an abnormal CC test. Women with an abnormal CC test also tended to have a familial history of early menopause. PMID- 14520192 TI - Intraoperative and postoperative morbidity associated with cystoscopy performed in patients undergoing gynecologic surgery. AB - OBJECTIVE: The purpose of this study was to evaluate the role of cystoscopy during major gynecologic surgery and to describe an interesting and rare complication that was encountered. STUDY DESIGN: This was an observational descriptive study. A retrospective review was performed. Data collected included intraoperative lower urinary tract injuries, injuries that were related directly to cystoscopy, and postoperative morbidity. RESULTS: One hundred one cases were evaluated. Eight lower urinary tract injuries were noted, 4 of which were detected at cystoscopy and 3 of which (2.97%) were detected intraoperatively before cystoscopy. There was one complication that was linked directly to cystoscopy. There were nine postoperative urinary tract infections. Total cost per case excluding anesthesia time was $54.42. CONCLUSION: Liberal use of cystoscopy in gynecologic surgery is recommended. The procedure is safe, easily learned, and inexpensive to perform. PMID- 14520194 TI - Adnexal masses and pregnancy: a 12-year experience. AB - OBJECTIVE: Our purpose was to describe pregnancy-associated adnexal masses in eastern North Carolina. STUDY DESIGN: A retrospective study was performed of 60 adnexal masses resected during pregnancy at a regional referral hospital from January 1990 to March 2002. RESULTS: Adnexal masses occurred in 0.15% of pregnancies. Average gestational age at diagnosis and surgery was 12 and 20 weeks, respectively. Fifty percent of ovarian tumors were mature cystic teratomas, 20% were cystadenomas, and 13% were functional ovarian cysts. Malignancy occurred in 13%. Tumors with low malignant potential comprised 63% of malignancies. Average cyst size was 11.5 cm for malignancies and 7.6 cm for benign lesions (P value <.05). The preterm birth rate was 9%, the miscarriage rate was 4.7% after elective cases, and average Apgar scores were 7.5 and 8.7 at 1 and 5 minutes. CONCLUSION: The incidence of malignancy in pregnancy-associated adnexal masses was high. Ultrasonography detected internal excrescences in the majority of tumors with low malignant potential. Fetal outcomes were not affected. PMID- 14520196 TI - Vaginectomy with pelvic herniorrhaphy for prolapse. AB - OBJECTIVE: The study was undertaken to report our experience with vaginectomy and pelvic herniorrhaphy for vaginal prolapse. STUDY DESIGN: This was an observational study of patients undergoing vaginectomy (n=41) or hysterovaginectomy (n=13) for stage III/IV vaginal prolapse. Morbidity was compared with cohorts who had undergone transvaginal repair of prolapse, by using the Mann-Whitney U test. RESULTS: Morbidity did not differ significantly (estimated blood loss) between the vaginectomy and hysterovaginectomy groups. There were no recurrent hernias (6-56 months). Operative time, estimated blood loss, and day of discharge were significantly greater for the posthysterectomy prolapse group compared with the vaginectomy group. Operative time was significantly greater for the uterovaginal prolapse group versus the hysterovaginectomy group. CONCLUSIONS: Vaginectomy with or without hysterectomy with pelvic herniorrhaphy is associated with a low rate of morbidity in a high risk patient population. Hysterovaginectomy is not associated with a clinically significant difference in morbidity over vaginectomy alone. Vaginectomy with or without hysterectomy should be offered as a surgical option to selected patients with severe genital prolapse. PMID- 14520198 TI - Correlation of symptoms with degree of pelvic organ support in a general population of women: what is pelvic organ prolapse? AB - OBJECTIVE: The purpose of this study was to evaluate the correlation between the symptoms of pelvic organ prolapse and the stage of support as determined by the pelvic organ prolapse quantification system. STUDY DESIGN: Four hundred ninety seven women who were seen for annual gynecologic examinations were recruited. Subjects underwent a pelvic examination and their degree of pelvic support was described according to the pelvic organ prolapse quantification system. They also completed a seven-question questionnaire regarding common symptoms of pelvic organ prolapse. Trend analysis was accomplished with linear regression. RESULTS: Only 477 subjects correctly responded to the questionnaire. They were aged 18 to 82 years (mean age, 44 years). Forty-seven percent were white, 52% were African American, and 1% were of another racial group. The number of subjects with the various pelvic organ prolapse quantification stages were stage 0 (18 subjects), stage I (214 subjects), stage II (231 subjects), and stage III (14 subjects). No subject had stage IV prolapse. The average number of positive responses per subject for the symptoms was 0.27 for stage 0, 0.55 for stage I, 0.77 for stage II, and 2.1 for stage III. This trend did not attain statistical significance. The correlation of symptoms with the leading edge of the prolapse revealed that the average number of symptoms that were reported per subject increased from <1 to >1 when the leading edge of the prolapse extended beyond the hymenal remnants. This trend was statistically significant. CONCLUSION: Women with pelvic organ prolapse with the leading edge of the prolapse beyond the hymenal remnants (some stage II and all stage III) have increased symptoms, which may help define symptomatic pelvic organ prolapse. PMID- 14520200 TI - Infectious urinary tract morbidity with prolonged bladder catheterization after radical hysterectomy. AB - OBJECTIVE: This study was undertaken to determine the incidence of catheter associated infection after radical hysterectomy and to evaluate the role of prophylactic antibiotics in these patients. STUDY DESIGN: A 4-year retrospective review of 102 women undergoing radical hysterectomy for cervical or endometrial cancer was performed. Clinical data were abstracted and analyzed with chi(2) and t tests. RESULTS: Catheter-associated infection was observed in 11% (12 of 102) and was not altered by the administration of prophylactic antibiotics (11.1% vs 11.8%, P=.95). Of the 12 women who had infection, 11 were treated as outpatients, and 1 patient required admission for pyelonephritis. Patient age, comorbid medical conditions, class of radical hysterectomy, perioperative complications, operative time, blood loss, catheter type, duration of catheterization, and length of hospitalization had no effect on the development of catheter-associated infection. CONCLUSION: The incidence of catheter-associated infection in women requiring prolonged catheterization after radical hysterectomy is relatively low. Withholding prophylactic antibiotics from these patients is a reasonable clinical option. PMID- 14520202 TI - Deciding on route of delivery for obese women with a prior cesarean delivery. AB - OBJECTIVES: This study was undertaken to estimate the vaginal birth after cesarean (VBAC) success rate, compare rates of infections in women attempting VBAC and those undergoing planned repeat cesarean, and compare the cost of these two plans of care for obese women. STUDY DESIGN: We performed a historical cohort analysis of singleton deliveries at >/=36 weeks' gestation in women with a body mass index 40 or greater and one prior cesarean delivery. Outcomes included rates of VBAC success and puerperal infections and mean cost of care. RESULTS: The cohort consisted of 122 mother-infant pairs, 61 each in the VBAC and cesarean groups. In the VBAC group, 57% (95% CI 45-70) of women were delivered vaginally. The VBAC group had higher rates of chorioamnionitis (13.1% vs 1.6%, P=.02), endometritis (6.6% vs 0%, P=.06), and composite puerperal infection (24.6% vs 8.2%, P=.01). Mean cost of care was similar for mothers ($4439 vs $4427, P=.95), infants ($1241 vs $1422, P=.49), and mother-infant pairs ($5680 vs $5851, P=.64). CONCLUSION: Compared with planned cesarean delivery, VBAC trials in obese women are three times as likely to be complicated by infection and do not result in reduced costs. PMID- 14520204 TI - Elevated plasma homocysteine in early pregnancy: a risk factor for the development of nonsevere preeclampsia. AB - OBJECTIVE: We have recently demonstrated that an elevated plasma homocysteine in early pregnancy is associated with the development of severe preeclampsia. The aim of this study was to determine whether an elevated plasma homocysteine in early pregnancy is also associated with the development of nonsevere preeclampsia. STUDY DESIGN: Blood was obtained from patients attending for a first antenatal visit. Subjects were asymptomatic women who subsequently developed nonsevere preeclampsia. Controls were matched for parity, gestational age, and date of sample collection. Plasma homocysteine was measured using fluorescence polarization immunoassay. RESULTS: There were 71 cases of nonsevere preeclampsia sampled at a mean gestational age (+/-SD) of 15.9+/-3.6 weeks and 142 controls at 15.6+/-3.4 weeks. The preeclampsia cases had a mean (+/-SD) homocysteine level of 8.4+/-2.4 micromol/L, whereas controls had a mean homocysteine of 7.07+/-1.5 micromol/L (P/=5 births. Odds ratios that were associated with parity were higher in women aged <60 years than in women aged >/=60 years. Age at first birth of >35 years was associated with a slight elevation in frequent leaking compared with women with age at first birth from 21 to 25 years but was stronger for women with age at first birth of <21 years (OR, 1.27; 95% CI, 1.13-1.42). CONCLUSION: In these women, leaking urine is common; this condition is most prevalent in white women, in older women, in parous women, and in women with a younger age at first birth. PMID- 14520213 TI - Are in-dwelling catheters necessary for 24 hours after hysterectomy? AB - OBJECTIVE: In-dwelling catheters for 24 hours after operation are used routinely in gynecologic surgery. This study assesses whether the immediate removal of an in-dwelling catheter after the operation affects the rate of recatheterization, febrile morbidity, symptomatic urinary tract infections, or subjective pain assessments. STUDY DESIGN: This study was a prospective randomized controlled trial comprised of 250 women who underwent hysterectomy and who did not require bladder suspension or strict fluid treatment. The in-dwelling catheter was removed either immediately after the operation or on the first day after the operation. The association between clinical variables and the length of catheterization were assessed by chi-squared analysis. RESULTS: Patients were assigned randomly into 2 groups, with no significant differences in the outcomes, only in the perception of pain. Clinical events included fever (>/=38.5 degrees C) that occurred in 6 patients in the in-dwelling catheter group compared with 5 patients in the early removal group (P=.01), symptomatic urinary tract infections in 3 patients in both groups (P=.99), and recatheterization in 3 patients in the in-dwelling catheter group compared with 5 patients in the early removal group (P=.17). Subjectively, patients in the early removal group reported significantly less pain than did the in-dwelling group (P<.001). CONCLUSION: The early removal of in-dwelling catheters after operation was not associated with an increased rate of febrile events, urinary tract infections, or need for recatheterization. In addition, subjective pain assessment was significantly less in the early removal group. Early removal of an in-dwelling catheter immediately after operation is not associated with adverse events. PMID- 14520214 TI - Double incontinence in urogynecologic practice: a new insight. AB - OBJECTIVES: This study was undertaken to evaluate the prevalence of anal incontinence in an urogynecologic setting and to investigate the relationship between lower urinary tract dysfunction and anal incontinence. STUDY DESIGN: The study included 504 women referred to our urogynecologic outpatient clinic who were prospectively investigated and asked specific questions on anal incontinence. Clinical and instrumental data were compared between women with urinary incontinence and with double incontinence, with further analysis for subgroups in the anal incontinent group of women (passive/urge). For continuous variables, the Wilcoxon rank sum test was used, and the Fisher exact test was applied to dicotomic variables. Logistic regression was used for categorical data. A level of P<.005 was considered significant. RESULTS: Of the investigated women, 20.2% were also anally incontinent. Women with double incontinence showed higher scores for urinary urgency (P=.010), which reached the established level of significance only in the subgroup with urge anal incontinence (P=.003). In this group, a higher prevalence of detrusor overactivity was observed. CONCLUSION: Anal incontinence is highly prevalent among women with lower urinary tract disorders. The existence of subgroups of patients having different kinds of anal and urinary disorders should be taken into consideration both for research purposes and for new treatment perspectives. PMID- 14520215 TI - The use of gonadotropin-releasing hormone antagonist in a flexible protocol: a pilot study. AB - OBJECTIVE: The purpose of this study was to investigate the efficacy of a flexible protocol of starting gonadotropin-releasing hormone antagonist according to the size of the leading follicle. STUDY DESIGN: This was a pilot study that included 123 couples who were undergoing in vitro fertilization/intracytoplasmic sperm injection cycles at the Egyptian IVF-ET Center. Couples were recruited into two groups: group I (n=64), gonadotropin-releasing hormone antagonist was administered when the diameter of the leading follicle reached 16 mm; group II (n=59), gonadotropin-releasing hormone antagonist was administered on day 6 of stimulation. RESULTS: The mean number of antagonist injections was significantly lower in the flexible protocol compared to the fixed protocol (3.4+/-1.1 vs 5.3+/ 1.8, P<.05). There was no significant difference between the two protocols regarding the number of embryos, implantation rate, clinical pregnancy rate (odds ratio, 0.85; 95% CI, 0.45-1.59) or multiple pregnancy rate (odds ratio, 1.26; 95% CI, 0.45-3.51). CONCLUSION: Starting the gonadotropin-releasing hormone antagonist according to the size of the leading follicle is as effective as starting on a fixed day and reduces the antagonist administration. PMID- 14520217 TI - Efficacy of thermal balloon ablation in patients with abnormal uterine bleeding. AB - OBJECTIVE: The purpose of this study was to assess changes in menstrual pattern, quality of life, and patient satisfaction after thermal balloon ablation for abnormal uterine bleeding. STUDY DESIGN: One hundred forty-one women who underwent thermal balloon ablation in our institution initially had their charts reviewed for demographics, procedure data, clinical history, and follow-up. Thereafter, a telephone interview was conducted to assess postprocedural menstrual pattern, quality of life, and patient satisfaction. Data were compared with the use of appropriate tests for categoric or continuous variables and logistic regression. RESULTS: The median follow-up time was 18 months, and a telephone interview was obtained for 119 of 141 patients. A reduction in days per cycle (9.6 vs 3.1 days, P<.0001) and in pads per day (12.8 vs 2.5 pads/d, P<.0001) and an improvement in self-reported quality of life scores (2.8 vs 9.0, P<.0001) were observed after thermal balloon ablation. Hysterectomy was required in 21 of 141 patients (15%). Assessment of the level of satisfaction showed that 96% of patients were satisfied or very satisfied with the procedure. No major complications or deaths were related to thermal balloon ablation. CONCLUSION: Thermal balloon ablation is a safe and efficient method to treat abnormal uterine bleeding. It reduces the menstrual flow, improves the quality of life, and remarkably fulfills expectations in selected patients. PMID- 14520216 TI - Effect of tubal sterilization on ovarian follicular reserve and function. AB - OBJECTIVE: Tubal ligation may reduce the ovarian blood flow and lead to tissue damage to the ovary. If so, this may also result in a significant decrease of the total follicular pool. We performed a long-term evaluation of ovarian reserve and function after tubal sterilization in a longitudinal prospective comparison cohort. STUDY DESIGN: In an university tertiary-care center, 26 women undergoing laparoscopic tubal sterilization with the use of bipolar coagulation, and 26 matched control subjects underwent measurement of follicle-stimulating hormone, luteinizing hormone, 17beta-estradiol, and inhibin on menstrual cycle day 3 before (baseline) and at 6, 12, 18, 24, and 60 months after the sterilization for ovarian reserve evaluation. At baseline and 12 and 24 months after tubal ligation, women who underwent sterilization were sampled every other day across an entire menstrual cycle for follicle-stimulating hormone, luteinizing hormone, 17beta-estradiol, inhibin, and progesterone determination to evaluate ovarian function. RESULTS: No significant changes were observed either within or between groups for any parameter, despite the fact that a 45% and 30% increase in follicle-stimulating hormone concentration from baseline to the 60-month control was detected in tubal sterilization and control groups of women, respectively. No significant changes were observed in the mean area under the curve of follicle stimulating hormone, luteinizing hormone, estradiol, inhibin, and progesterone per menstrual cycle at baseline and 12 and 24 months after sterilization. CONCLUSION: This 5-year follow-up study suggests that there is neither an accelerated decline of ovarian follicular reserve nor ovarian dysfunction after tubal sterilization by electrocoagulation. PMID- 14520219 TI - Depressive symptoms among women with vulvar dysesthesia. AB - OBJECTIVE: The purpose of this study was to determine whether vulvar dysesthesia is associated with elevated depressive symptoms. STUDY DESIGN: This was a cross sectional case-control study of women who underwent treatment of vulvar dysesthesia (n=32) or who were seen for a routine gynecologic examination (n=32). Subjects completed measures of depressive symptoms and pain and a sexual and medical history. Multivariate and univariate analyses were conducted. RESULTS: Analyses that were adjusted for age, education, and medical conditions indicated that vulvar dysesthesia was associated positively with depressive symptoms (P=.002). However, this was attributable to the somatic (P=.002) rather than cognitive-affective symptoms (P=.16) of depression, partially related to the endorsement of sexual disinterest, and mediated by pain reports. CONCLUSION: Vulvar dysesthesia is associated with elevated depressive symptom severity, although not to the extent that indicates probable depressive disorder. In this condition, depressive symptoms are likely to be a measurement artifact, rather than a depressive process. Certain depressive symptoms (eg, sexual disinterest) directly inflate depression estimates in this patient group. PMID- 14520218 TI - Estrogen receptor expression in vulvar vestibulitis syndrome. AB - OBJECTIVE: A pilot study was performed to investigate the relationship between vulvar vestibulitis syndrome and estrogen receptor expression. STUDY DESIGN: Women with a diagnosis of vulvar vestibulitis syndrome had tissue samples taken for vulvar estrogen receptor-alpha expression and this was compared with a control group. RESULTS: The study group showed a significant decrease in estrogen receptor expression, and 50% of the samples did not exhibit any receptor expression. CONCLUSION: There appears to be a subgroup of women with vulvar vestibulitis syndrome who exhibit abnormal estrogen receptor-alpha expression. This may be helpful in explaining why some women are resistant to medical treatment and may allow treatment to be prescribed more effectively. PMID- 14520220 TI - Factors that affect the quality of cytologic cervical cancer screening along the Mexico-United States border. AB - OBJECTIVE: The purpose of this study was to identify the factors that affect the quality of cytologic cervical cancer screening conducted in public sector clinics in Sonora, Mexico. STUDY DESIGN: We assessed the quality of cervical cytology before and after a 2-hour training session and the implementation of cervical spatulas and endocervical brushes. Additionally, we conducted a cross-sectional study of reproductive aged women who attended public clinics in paired border communities in Sonora and Arizona. Cervical cytologic specimens (n=2436) were collected and compared for adequacy and cytologic diagnosis and for the prevalence of human papillomavirus infection. RESULTS: The training intervention achieved significantly improved the rates of satisfactory but limited smears and unsatisfactory smears. The cross-sectional study revealed comparable quality indicators for cytologic specimens that were collected in Sonora and Arizona clinics. CONCLUSION: A high-quality cytology-based cervical cancer screening program is possible in public sector clinics in Sonora, with indicators similar to those achieved in comparable Arizona clinics. PMID- 14520221 TI - Initiating contraception in sexually transmitted disease clinic setting: a randomized trial. AB - OBJECTIVE: Our purpose was to determine the effectiveness of sexually transmitted disease (STD) clinic-initiated contraceptive care. STUDY DESIGN: Nonpregnant women (n=877) attending an urban STD clinic using either no contraception or only condoms were randomly assigned to either an intervention (n=437) or control group (n=440). Both groups received condoms with spermicide and a referral list of primary care providers (PCP) for ongoing reproductive health care, and the intervention group also received enhanced contraceptive counseling, initial provision of contraception, and facilitated referral to a PCP. Outcomes measured at 4-, 8-, and 12-month follow-up were transition to a PCP, effective contraceptive use (ECU), interval pregnancy, and STD. RESULTS: The median time to PCP transition was 79 days for the intervention group versus 115 days for the control group (P=.007). Rates of ECU were higher for the intervention group than for control group at the 4-month visit (50% vs 22%, P<.0001) as well as the 8 month visit, although in the intervention group ECU diminished over the course of the study. During follow-up, pregnancy outcomes were documented for 229 women (26.1%), for an overall pregnancy rate of 38.2 per 100 person-years of follow-up. Of the 159 pregnancies defined by patient self-report, 153 (96.2%) were described as unintended and 32 (20%) resulted in a therapeutic abortion. The pregnancy rate was 15% lower in the intervention (105/437, 24.0%) than the control group (124/440, 28.2%) (P=.16), but this difference was not statistically significant. CONCLUSION: The intervention helped women transition to a PCP and initiate ECU but did not significantly reduce the pregnancy rate. More intensive interventions are needed to prevent unintended pregnancy in this high-risk population. PMID- 14520222 TI - Fetal myelomeningocele repair: short-term clinical outcomes. AB - OBJECTIVE: The study was undertaken to evaluate short-term clinical outcomes with antenatal myelomeningocele (MMC) repair. STUDY DESIGN: Retrospective review of 50 fetuses that underwent open fetal MMC closure was performed. Inclusion criteria included less than 26 weeks' gestation, thoracic to S1 level defect, absent clubfoot deformity, Arnold-Chiari malformation, ventriculomegaly less than 17 mm, normal karyotype, and no other anomalies. RESULTS: Perinatal survival was 94% (47/50 fetuses). Mean age at delivery was 34 weeks 3 days. All fetuses demonstrated reversal of hindbrain herniation. Forty-three percent of the 50 fetuses have required ventriculoperitoneal shunting compared with 100% thoracic, 88% lumbar, and 68% sacral (85% overall) in 297 historic controls. Better-than predicted leg function was seen in 57% of thoracic and lumbar level lesion patients. CONCLUSION: Early experience with fetal MMC repair suggests a decreased need for ventriculoperitoneal shunting, arrest or slowing of progressive ventriculomegaly, and consistent resolution of hindbrain herniation. However, further long-term follow-up is needed to evaluate neurodevelopment and bladder and bowel function. PMID- 14520223 TI - The natural history of antenatal nuchal cords. AB - OBJECTIVE: The purpose of this study was to test the null hypotheses that nuchal cords occur with equal frequency throughout gestation, the presence of an antenatal nuchal cord is usually a random or chance event, nuchal cords do not usually persist, and nuchal cords do not usually produce evidence of acute fetal compromise before labor. STUDY DESIGN: A prospective subject-, examiner-, and physician-blinded study design was used. The fetuses were examined serially at 24 to 26, 30 to 32, and 36 to 38 weeks' gestation and during labor and delivery. Measurements included: ultrasonic or visual detection of nuchal cords and assessment of fetal and neonatal well-being by using standard clinical techniques. RESULTS: The incidence of nuchal cord rose with advancing gestation from 12% at 24 to 26 weeks to 37% at term. However, at each gestational age, its occurrence was a random or chance event and was not associated with clinical evidence of fetal compromise before labor. CONCLUSION: Antenatal nuchal cords usually occur randomly with increased frequency in late gestation and appear to be a normal part of intrauterine life that is rarely associated with perinatal morbidity and mortality. PMID- 14520224 TI - Weight at birth and subsequent risk of preeclampsia as an adult. AB - OBJECTIVE: We examined the influence of maternal birth weight on the risk of the development of preeclampsia, a likely precursor to adult chronic disease. STUDY DESIGN: This hospital-based case-control study included 181 preeclampsia cases and 349 control subjects. Participants provided information about their birth weight and other covariates that included medical and reproductive history, prepregnancy weight, and adult height. Odds ratios and 95% CIs were estimated by logistic regression. RESULTS: The risk of preeclampsia decreased as maternal birth weight increased (P=.01). After an adjustment was made for confounders, data showed that women with a low birth weight (<2500 g) had a 2.3-fold increased risk of experiencing preeclampsia (95% CI, 1.0-5.3) as compared with women who weighed 2500 to 2999 g at birth. Conversely, women with a birth weight of >/=4000 g appeared to have a nonstatistically significant, but >50%, reduction in the risk of experiencing preeclampsia (95% CI, 0.2-1.2). This relationship differed for lean and overweight women (body mass index, <25 kg/m(2) vs >/=25 kg/m(2)). Among lean women, those who were low birth weight had a near doubling in risk of the development of preeclampsia (odds ratio, 1.9; 95% CI, 0.8-4.6), although this association did not reach statistical significance. However, among overweight women, those women who weighed <2500 g at birth had an almost 4-fold increased risk of experiencing preeclampsia (odds ratio, 3.8; 95% CI, 1.1-13.8). CONCLUSION: These results confirm two earlier reports and expand the literature by showing that women who are small at birth and who become overweight as adults are at particularly high risk of the development of preeclampsia. PMID- 14520225 TI - Does labor influence neonatal and neurodevelopmental outcomes of extremely-low birth-weight infants who are born by cesarean delivery? AB - OBJECTIVE: The purpose of this study was to examine the influence of labor on extremely-low-birth-weight infants who were born by cesarean delivery with reference to neonatal and neurodevelopmental outcomes. We hypothesized that infants who are born by cesarean delivery without labor will have better outcomes than those infants who are born by cesarean delivery with labor. STUDY DESIGN: This was a retrospective cohort study of extremely-low-birth-weight infants (birth weight, 401-1000 g) who were born by cesarean delivery and cared for in the National Institute for Child Health and Human Development Neonatal Network, during calendar years 1995 to 1997. A total of 1606 extremely-low-birth-weight infants were born by cesarean delivery and survived to discharge. Of these, 1273 infants (80.8%) were examined in the network follow-up clinics at 18 to 22 months of corrected age and had a complete data set (667 infants were born without labor, 606 infants were born with labor). Outcome variables that were examined include intraventricular hemorrhage grade 3 to 4, periventricular leukomalacia, and neurodevelopmental impairment. RESULTS: Mothers in the cesarean delivery without labor group were older (P<.001), more likely to be married (P<.05), less likely to be supported by Medicaid (P<.01), more likely to have preeclampsia/hypertension (P<.001), more likely to receive prenatal steroids (P<.005), and less likely to have received antibiotics (P<.001). Infants who were born by cesarean delivery without labor had higher gestational age (P<.001), lower birth weight (P<.01), and were less likely to be outborn (P<.001). By univariate analysis, infants who were born by cesarean delivery with labor had a higher incidence of grade 3 to 4 intraventricular hemorrhage (23.3% vs 12.1%, P<.001), periventricular leukomalacia (8.5% vs 4.7%, P<.02), and neurodevelopmental impairment (41.7% vs 34.6%, P<.02). Logistic regression analysis that controlled for all maternal and neonatal demographic and clinical variables that were statistically associated with labor or no labor revealed that the significant differences in grade 3 to 4 intraventricular hemorrhage, periventricular leukomalacia, and neurodevelopmental impairment were no longer evident. CONCLUSION: In extremely-low-birth-weight infants who were born by cesarean delivery and after control for other risk factors, labor does not appear to play a significant role in adverse neonatal outcomes and neurodevelopmental impairment at 18 to 22 months of corrected age. PMID- 14520226 TI - Preprandial versus postprandial blood glucose monitoring in type 1 diabetic pregnancy: a randomized controlled clinical trial. AB - OBJECTIVE: This study was undertaken to compare preprandial and postprandial capillary glucose monitoring in pregnant women with type 1 diabetes. STUDY DESIGN: Sixty-one women with type 1 diabetes were randomly assigned at 16 weeks' gestation to preprandial or postprandial blood glucose monitoring using memory based glucose reflectance meters throughout pregnancy. Serial measurements of hemoglobin A1c and fructosamine were obtained throughout pregnancy. Insulin, glucose, and insulin-like growth factor-I (IGF-I) were measured in cord blood at delivery. Neonatal anthropometric measures were performed within 72 hours of delivery RESULTS: Maternal age, parity, age of onset of diabetes, number of prior miscarriages, smoking status, social class, weight gain in pregnancy, and compliance with therapy were similar in the two groups. The postprandial monitoring group had a significantly reduced incidence of preeclampsia (3% vs 21%, P<.048), a greater success in achieving glycemic control targets (55% vs 30%, P<.001) and a smaller neonatal triceps skinfold thickness (4.5+/-0.9 vs 5.1+/-1.3, P=.05). CONCLUSION: Postprandial capillary blood glucose monitoring in type 1 diabetic pregnancy may significantly reduce the incidence of preeclampsia and neonatal triceps skinfold thickness compared with preprandial monitoring. PMID- 14520227 TI - Pregnancy outcome in spontaneous twins versus twins who were conceived through in vitro fertilization. AB - OBJECTIVE: The purpose of this study was to compare maternal and neonatal complications in spontaneous versus in vitro fertilization twins. STUDY DESIGN: Twin gestations that were delivered from 1995 to 2000 were reviewed. Cases consisted of 56 in vitro fertilization twins, each of which was matched to two control mothers by age and parity. They were compared regarding various maternal and neonatal complications. RESULTS: In vitro fertilization twins were more likely to have preterm labor compared with control twins, with no difference in the incidences of pregnancy-induced hypertension, gestational diabetes mellitus, placenta previa, or preterm premature rupture of membranes between the two groups. The cesarean delivery rate was significantly higher in cases of twins who were conceived by in vitro fertilization (76.8% vs 58.0%, P=.026), despite a similar rate of elective cesarean delivery and the incidence of nonvertex twin A in both groups. The preterm delivery rate was significantly higher (67.9% vs 41.1%, P=.002) and the gestational age was significantly lower (35+/-3 weeks vs 36+/-3 weeks, P=.043) in cases compared with control subjects. Both twins were, on the average, 230 g lighter in the in vitro fertilization group compared with the control group. However, intrauterine growth restriction was more frequent in the control group (36.6% vs 25%, P=.044). There was a significantly higher incidence of admission to the neonatal intensive care unit, respiratory distress syndrome, a need for mechanical ventilation, and pneumothorax in cases compared with control subjects. CONCLUSION: When compared with spontaneous twins, in vitro fertilization twins are more likely to be delivered by cesarean delivery and to have a higher incidence of preterm birth and prematurity-related respiratory complications with a longer nursery stay. PMID- 14520229 TI - Helicobacter pylori infection and the occurrence and severity of gastrointestinal symptoms during pregnancy. AB - OBJECTIVE: We investigated the possible role of Helicobacter pylori infection in the occurrence and severity of gastrointestinal symptoms during pregnancy in a large group of mothers after delivery. STUDY DESIGN: Between November 2000 and November 2001, mothers were recruited after delivery at the Department of Gynecology and Obstetrics at the University of Ulm. Present H pylori infection was determined by (13)C-urea breath test. Associations between gastrointestinal symptoms during pregnancy (sickness, vomiting, increased saliva production, heartburn) and H pylori infection were quantified by crude and adjusted odds ratios with 95% CI. RESULTS: Twenty-three percent of the 898 mothers had a current H pylori infection. Eighty-four percent of the mothers reported at least one of the evaluated gastrointestinal symptoms, and 30% of the mothers reported at least one physician visit because of the severity of these symptoms. None of the analyzed gastrointestinal symptoms showed an association to a current H pylori infection after an adjustment for the covariates, even after a virulence marker of H pylori infection was taken into account. CONCLUSION: This study does not support an involvement of H pylori infection in the generation of gastrointestinal symptoms during pregnancy. PMID- 14520228 TI - Vitamin C intake and the risk of preterm delivery. AB - OBJECTIVE: Ascorbic acid deficiency may lead to premature rupture of the membranes. STUDY DESIGN: The study included a prospective cohort of pregnant women, aged >/=16 years, with singleton gestations who received care at one of four prenatal clinics in central North Carolina from 1995 through 1998. Vitamin C intake preconceptionally and during the second trimester was examined for its association with preterm delivery and subsets of preterm labor, premature rupture of the membranes, and medical induction in 2064 women. RESULTS: Women who had total vitamin C intakes of <10th percentile preconceptionally had twice the risk of preterm delivery because of premature rupture of the membranes (relative risk, 2.2; 95% CI, 1.1, 4.5). This risk was attenuated slightly for second-trimester intake (relative risk, 1.7; 95% CI, 0.8, 3.5). The elevated risk of preterm premature rupture of the membranes was greatest for women with a low vitamin C intake during both time periods. CONCLUSION: Because diet and supplement use are modifiable behaviors, corroboration of these findings would suggest a possible intervention strategy. PMID- 14520230 TI - Evaluation of quantitative ultrasound tissue characterization of the cervix and cervical length in the prediction of premature delivery for patients with spontaneous preterm labor. AB - OBJECTIVE: This study was to evaluate the predictive value of the uterine cervix tissue with the use of quantitative ultrasound gray level analysis for preterm delivery. STUDY DESIGN: Sixty-eight patients with preterm labor between 20 and 35 weeks of gestation were included. When two-dimensional transvaginal ultrasound measurement of cervical length was completed, a region of interest of constant size was defined in the midsection of the posterior wall, and the tissue-specific gray scale was determined. Preterm delivery of <37 weeks of gestation was sought. RESULTS: Twenty-eight patients (41.2%) were delivered preterm. The risk for preterm delivery was increased significantly in patients with cervical length of /=4 (odds ratio, 3.44; 95% CI, 1.21-9.75), and with decreased mean gray scale value (odds ratio, 12.13; 95% CI, 3.69-39.88). Parity and uterine contractions were not significant as predictors for preterm delivery, although the risk for preterm delivery increased with higher parity (odds ratio, 1.8; 95% CI, 0.68-4.79). The risk for preterm delivery remained nearly the same by uterine contractions (odds ratio, 0.92; 95% CI, 0.28-3.01). A mean scale value of /=30 eosinophils per high-power field) were included in a retrospective review. RESULTS: A diagnosis of eosinophilic esophagitis was made in 31 patients (24 men, 7 women; mean age 34 years, range 14-77 years). The diagnosis was made in 19 (61%) of the 31 patients during the most recent 2 years (none between 1981 and 1994 vs. 12 between 1995-2000 vs. 19 between 2000-2001). Esophageal mucosal furrows were present in 30 (97%), a finding infrequently recognized before 2001. Dysphagia was documented in 26 (89%). Symptoms had been present for long periods before diagnosis (mean 54 months; range 0-180 months), and diagnosis was delayed in 7 (mean 81 months, range 20-144 months) because sentinel features were overlooked at endoscopy. Strictures, often evident only as a result of mucosal shearing during dilation, were present in 17 (57%). Esophageal dilation preformed in 17 (mean 3.4 dilations per patient, range 1-13) consistently relieved symptoms; tears were recorded in 13 (87%), but no serious complication resulted. CONCLUSIONS: Eosinophilic esophagitis in adults of all ages is more common than recognized. Mucosal furrows are easily overlooked at endoscopy although this finding is an important clue to diagnosis. Strictures, a frequent consequence, can be safely managed by dilation. PMID- 14520284 TI - Low-dose flunitrazepam for conscious sedation for EGD: a randomized double-blind placebo-controlled study. AB - BACKGROUND: EGD with conscious sedation is a safe procedure, but complications such as hypoxia can occur. The efficacy and safety of low-dose flunitrazepam (0.25 mg) was compared with a standard dose of flunitrazepam (0.5 mg) for moderate sedation during EGD. METHODS: In a randomized, double-blind, placebo controlled trial, 75 outpatients (40 men, 35 women, mean age 45 [11] years) undergoing screening EGD were randomly assigned to one of 3 treatment arms: 0.25 mg of flunitrazepam (F0.25 group), 0.5 mg of flunitrazepam (F0.5 group), or placebo (normal saline solution), each administered intravenously. Patient tolerance was scored by using self-assessment questionnaires with visual analogue scales. Cardiopulmonary complications were assessed by monitoring blood pressure, heart rate, oxygen saturation, and the electrocardiogram during the procedure. RESULTS: The patient tolerance scores in the F0.25 and F0.5 groups, respectively 2.1 (2.1) and 2.3 (2.5), were significantly lower than that for the placebo group (6.5 [3.0]); there was no significant difference between F0.25 and F0.5. Cardiopulmonary complications in the F0.25 group were significantly lower than in the F0.5 group. Oxygen desaturation (oxygen saturation < 90%) was noted in two of 25 patients in the F0.5 group. Post-procedure drowsiness was observed in two of 24 (8.3%) patients in the F0.25 group and 3 of 21 (14.3%) in the F0.5 group (p = 0.2438). CONCLUSIONS: Patient tolerance of EGD with low-dose flunitrazepam (0.25 mg intravenously) was similar to that with a standard dose (0.5 mg intravenously) and significantly better than in the placebo group. Oxygen desaturation was observed only in the group that received the standard dose, suggesting that sedation with low-dose flunitrazepam is efficacious and safe for EGD. PMID- 14520285 TI - EUS detection of pancreatic endocrine tumors in asymptomatic patients with type 1 multiple endocrine neoplasia. AB - BACKGROUND: EUS is highly sensitive, specific, and cost-effective for localization of pancreatic neuroendocrine tumors. EUS screening of asymptomatic patients with multiple endocrine neoplasia type 1 has not been described. METHODS: EUS was used to evaluate all patients with known or suspected pancreatic neuroendocrine tumors. Asymptomatic patients with either a confirmed genetic or clinical diagnosis of multiple endocrine neoplasia type 1 were evaluated with EUS. The results were correlated with surgical and histopathologic findings. RESULTS: A total of 65 patients with multiple endocrine neoplasia type 1 underwent 132 EUS procedures over an 8-year period, including 13 asymptomatic patients. Five of the 13 asymptomatic patients had normal serum gastrin levels, and 8 had levels less than 4 times the upper limit of normal. EUS demonstrated pancreatic neuroendocrine tumors in 11 of these patients, ranging in size from 0.4 to 4 cm (mean 10 mm). Ten of the 11 patients with tumors at EUS underwent surgical exploration, and 23 of 28 (82%) tumors removed surgically had been identified by EUS. At a mean follow-up of 44 months, no liver or lymph node metastases were demonstrated in any patient. CONCLUSIONS: In this first cohort study of asymptomatic patients with multiple endocrine neoplasia type 1 undergoing screening EUS, pancreatic neuroendocrine tumors were identified before the development of significant biochemical test abnormalities. Aggressive early surgical treatment may improve the prognosis for these patients. PMID- 14520286 TI - Preoperative diagnosis and staging of gallbladder carcinoma by EUS. AB - BACKGROUND: EUS has recently been shown to be efficacious for the preoperative assessment of depth of invasion of gallbladder carcinoma. This study assessed the value of EUS for determining T stage (International Union Against Cancer). METHODS: Preoperative EUS findings in 41 patients with gallbladder carcinoma were analyzed retrospectively. EUS images were classified according to the shape of the tumor and the adjacent gallbladder wall structure as follows: type A, pedunculated mass with preserved adjacent wall structures; type B, sessile and/or broad-based mass with a preserved outer hyperechoic layer of the gallbladder wall; type C, sessile and/or broad-based mass with a narrowed outer hyperechoic layer; type D, sessile and/or broad-based mass with a disrupted outer hyperechoic layer. EUS and histopathologic findings were compared, including the depth of invasion of the tumor in the resection specimen. RESULTS: The 4 categories of EUS images of gallbladder carcinoma correlated with the histologic depth of invasion and T stage. Accuracies for the EUS classification as type A corresponding to pTis, type B to pT1, type C to pT2, and type D to pT3-4 were, respectively, 100%, 75.6%, 85.3%, and 92.7%. CONCLUSIONS: Preoperative EUS imaging accurately depicts T stage of gallbladder carcinoma and allows for effective therapeutic decision making. PMID- 14520287 TI - EUS-FNA of recurrent postoperative extraluminal and metastatic malignancy. AB - BACKGROUND: EUS-guided FNA is safe and accurate for the diagnosis of benign or malignant neoplasia and lymphadenopathy; however, its role in the diagnosis of recurrent malignancy is not well described. METHODS: A prospectively updated EUS guided FNA cytology database was used to identify patients in whom a diagnosis of postoperative, recurrent, extraluminal, or metastatic malignancy was made over a 5-year period. Only patients with a positive EUS-guided FNA were included in the analysis. All had undergone surgery for the primary malignancy and were in clinical and/or radiographic remission before the initial suspicion of tumor recurrence. RESULTS: Twenty-one patients underwent EUS-guided FNA of 21 lesions (19 masses, 2 lymph nodes) because of a suspicion of recurrent malignancy based on CT (n = 17) or EUS (n = 4) findings. Median time from the initial diagnosis to recurrence was 26 months (range 5-276 months). Lesions were located in the pancreas (9 patients), mediastinum (7), liver (3), perigastric region (1), and liver hilum (1). EUS-guided FNA (mean number of needle passes, 4.5; range 2-8) obtained diagnostic material for recurrent malignancy in all patients as follows: esophageal (6 patients), renal cell (6), pancreatic (2), breast (2), colon (2), bile duct (1), Ewing's sarcoma (1), and lung (1) cancer. No complication was encountered. Transgastric EUS-guided FNA (4 patients), distal, or transesophageal EUS-FNA (2) proximal to a surgical anastomosis was required to confirm recurrence in all 6 patients with esophageal cancer. The initial cytologic diagnosis of recurrent malignancy was made by EUS in 20 of 21 (95%) patients. One patient with recurrent breast cancer had CT-guided FNA of a right lung mass preceding EUS guided FNA of an AP window lymph node. CONCLUSIONS: EUS-guided FNA can detect and safely diagnose recurrent malignancy in the mediastinum, retroperitoneum, and liver. When possible, correlation between EUS-guided FNA cytology and original tumor histopathology/cytology, or the use of immunostaining to confirm the diagnosis, is recommended. PMID- 14520288 TI - The impact of post-procedure interpretation of ERCP X-ray films by radiologists on patient care: should it be routine or selective? AB - BACKGROUND: Review of ERCP x-ray films by radiologists is routine, but the utility of this practice is unproven. The aim of this study was to assess whether the routine post-procedural interpretation of ERCP films by radiologists alters patient management. METHODS: A retrospective analysis of 212 ERCPs followed by a prospective analysis of 112 ERCPs was performed. Comparative ductogram interpretations were categorized as: I, complete agreement; II, minor findings reported only by the radiologist; III, findings reported only by the endoscopist; and IV, major findings reported only by the radiologist that altered or should have altered management. RESULTS: In the retrospective analysis, 289 ductograms were identified, and interpretations were classified as: category I, 73%; category II, 16%; category III, 10.7%; and category IV, 0.3%. In the prospective study, interpretations of 167 ductograms were analyzed and classified as follows: category I, 84%; category II, 11%; category III, 5%; category IV, none. CONCLUSIONS: Post-procedure interpretation of ERCP spot x-ray films by radiologists adds little to patient management. Selective consultation with radiologists would appear to be more appropriate than review by radiologists of ERCP spot x-ray films on a routine basis. PMID- 14520289 TI - Perforation during colonoscopy in endoscopic ambulatory surgical centers. AB - BACKGROUND: Perforation as a complication of colonoscopy is estimated to occur in 0.01% to 0.3% of procedures, but the frequency in ambulatory settings is unknown. This study determined the number of perforations occurring within a network of endoscopic ambulatory surgery centers. METHODS: A total of 116,000 colonoscopies were performed within one network of 45 endoscopic ambulatory surgery centers in the United States during 1999. All identified perforations were reported to the network clinical director and reviewed by a panel of 3 gastroenterologists. RESULTS: There were 37 (0.03%) perforations; 27 in women and 10 in men. Median patient age was 75 years (range 39-87 years); 18 patients (49%) had diverticular disease and 20 (54%) had a history of pelvic or colonic surgery. Twenty-four (65%) procedures were diagnostic, and 13 (35%) were therapeutic. The most common site of perforation was the sigmoid colon (62%); followed by the ascending colon (16%); cecum, transverse colon, and splenic flexure (11%); and rectum, anastomotic, or unknown (11%). The time to diagnosis ranged from immediate to 72 hours (29 <1 hour, 8 >1 hour). All patients were hospitalized; 35 (95%) underwent exploratory laparotomy, and 2 (5%) were treated conservatively. No patient died. CONCLUSIONS: Reported perforations for procedures performed in endoscopic ambulatory surgery centers occurred most frequently during diagnostic colonoscopy in older woman with a history of surgery or diverticular disease. Reported perforations in endoscopic ambulatory surgery centers were uncommon. PMID- 14520290 TI - Primary prophylaxis of variceal bleeding. PMID- 14520291 TI - Sedation with droperidol for sphincter of Oddi manometry. PMID- 14520292 TI - Does the addition of endoscopic sphincterotomy to stent insertion improve drainage of the bile duct in acute suppurative cholangitis? PMID- 14520293 TI - Staple impaction in cervical esophagus. PMID- 14520294 TI - Cholecystoduodenal fistula. PMID- 14520295 TI - Lymphangioma of small intestine. PMID- 14520296 TI - Insulinoma: identification by EUS and intraoperative US. PMID- 14520297 TI - Ascending colon ameboma. PMID- 14520298 TI - Pseudomelanosis of stomach, duodenum, and jejunum. PMID- 14520299 TI - Colonoscopy with the variable stiffness colonoscope. PMID- 14520300 TI - Transluminal endosurgery: single lumen access anastomotic device for flexible endoscopy. AB - BACKGROUND: Forming anastomoses between two hollow organs at flexible endoscopy might reduce the need for surgery for obstructing malignancy. Current methods require access to both lumens. The aim of this work was to develop methods of forming an anastomosis at flexible endoscopy, such as a gastrojejunostomy or cholecystoduodenostomy, when access to only one lumen is feasible. METHODS: A modified needle was passed through a large-channel echoendoscope from the accessible lumen into the target hollow organ. An anastomotic device was formed by using two 7F catheter segments, which were pushed over a guidewire into the target, the less accessible lumen. When released, by withdrawing the guidewire, the catheters formed a cross shape and created an anastomosis when compressed against a plate from the accessible side. OBSERVATIONS: These devices were tested in live animal experiments. With an echoendoscope in the stomach, it was repeatedly possible to place needles, threaded tags, and guidewires into the small intestine and gallbladder. In 4 to 7 days, anastomoses were formed in 16 pigs between the small intestine and the stomach, and between the gallbladder and the stomach. The initial diameter of the anastomoses ranged from 3 to 9 mm. No complication occurred. CONCLUSIONS: It is feasible to form anastomoses at flexible endoscopy when access is limited to a single side. PMID- 14520301 TI - In vivo Doppler optical coherence tomography of mucocutaneous telangiectases in hereditary hemorrhagic telangiectasia. AB - BACKGROUND: Hereditary hemorrhagic telangiectasia is characterized by mucocutaneous telangiectases and visceral arteriovenous malformations. Knowledge is limited concerning the development hemodynamics of mucocutaneous telangiectases. Doppler optical coherence tomography can demonstrate microvascular blood flow at flow rates as low as 20 microm/second, which is up to approximately 100 times more sensitive than Doppler US. The aims of this study were to collect in vivo Doppler optical coherence tomography images of mucocutaneous telangiectases and normal surrounding mucosa and skin, and to gain experience for an in vivo GI endoscopic study. It was hypothesized that visibly normal areas may have occult telangiectases and that mucocutaneous telangiectases that have bled may have a higher rate of blood flow than mucocutaneous telangiectases with no history of bleeding. METHODS: Twelve patients with hereditary hemorrhagic telangiectasia and mucocutaneous telangiectases were studied. Two to 3 visible mucocutaneous telangiectases on the digits, lips, and tongue were imaged with Doppler optical coherence tomography, along with visually normal surrounding areas at each site. The Doppler optical coherence tomography images were obtained in 0.5 second by using 1310 nm light. RESULTS: A total of 67 mucocutaneous telangiectases from the 12 patients were imaged (38 digit, 16 lip, 13 tongue). Blood flow was demonstrated within every mucocutaneous telangiectasis imaged. Doppler optical coherence tomography did not identify any abnormal vasculature within visually normal areas. Mucocutaneous telangiectases with a history of bleeding (n = 18) were situated closer to the surface, compared with mucocutaneous telangiectases with no bleeding history (n = 49), but there was no difference in the Doppler flow appearance. CONCLUSIONS: Visually normal areas in patients with hereditary hemorrhagic telangiectasia did not appear to have abnormal vasculature. Mucocutaneous telangiectases with a history of bleeding were more superficial but were otherwise similar to mucocutaneous telangiectases with no bleeding history. PMID- 14520302 TI - Capsule endoscopy regional transit abnormality: a sign of underlying small bowel pathology. AB - BACKGROUND: Wireless capsule endoscopy is being used increasingly to investigate GI bleeding of obscure etiology and other small bowel abnormalities. Currently, there is no standard for capsule endoscopy image interpretation and classification METHODS: This report describes an abnormality, termed "regional transit abnormality," noted during a review of capsule endoscopic images. Capsule regional transit abnormality simply refers to delayed capsule transit within a segment of small bowel, with or without visualization of a mucosal abnormality. The slowed transit usually is accompanied by mucosal collapse, sometimes with the appearance of the capsule pressing or impacting upon the mucosa. Regional transit abnormality usually persists more than 15 minutes CONCLUSIONS: Capsule regional transit abnormality is considered to be a "red flag" sign that indicates a likelihood of some underlying small bowel pathology. PMID- 14520303 TI - Endoscopic management of biliary obstruction caused by cavernous transformation of the portal vein. AB - BACKGROUND: Symptomatic biliary obstruction caused by cavernous transformation of the portal vein is an extremely rare disorder for which there is no consensus as to optimal treatment. The results of endoscopic treatments in a small group of patients is reviewed. METHODS: A total of 10 patients (5 men, 5 women; mean age 36.1 years, range 17-48 years) with severe biliary strictures were treated between 1995 and 2001. Biliary sphincterotomy was performed in all patients. Four patients also underwent balloon dilation, nasobiliary drainage, and stone or sludge extraction by using a balloon. All patients had stent insertion. OBSERVATIONS: The mean duration of therapy was 3.3 years (range 1-7 years). There was no complication directly related to the endoscopic procedures except for hemobilia that occurred in one patient during stent removal. Cholangitis developed in 5 patients during the therapy period and was treated endoscopically. In 4 patients, significant improvement in the biliary stricture was observed and stents were removed in 3. These patients were followed without stent insertion for one year. CONCLUSIONS: Endoscopic management of biliary stricture caused by cavernous transformation of the portal vein appears to be effective and safe. PMID- 14520304 TI - Successful early treatment of Boerhaave's syndrome by endoscopic placement of a temporary self-expandable plastic stent without fluoroscopy. PMID- 14520305 TI - EUS-guided drainage of a diverticular abscess as an adjunct to surgical therapy. PMID- 14520306 TI - Gamma/delta T-cell lymphoma of duodenal bulb: a case report. PMID- 14520307 TI - Choledochal cyst mimicking a pancreatic pseudocyst: case report and review. PMID- 14520308 TI - Suture-induced dysphagia: the role of endoscopy in diagnosis and therapy. PMID- 14520309 TI - Unusual presentations of hepatic clonorchiasis. PMID- 14520310 TI - Colonoscopy practice variation. PMID- 14520311 TI - Management of rectal carcinoid tumors. PMID- 14520315 TI - Interruption of cardiopulmonary resuscitation with the use of the automated external defibrillator in out-of-hospital cardiac arrest. AB - STUDY OBJECTIVE: The protocol for the use of the automated external defibrillator calls for a period of "hands-off" time, during which no cardiopulmonary resuscitation (CPR) can be performed. We assessed the actual interruption time of CPR during the use of the automated external defibrillator in patients in out-of hospital cardiac arrest. METHODS: This study included 184 patients experiencing out-of-hospital cardiac arrest in which an automated external defibrillator was applied by first responders. ECG and voice recordings from the automated external defibrillator were downloaded and analyzed. Start and end times of CPR were recorded, as were intervals measured from the recordings concerning the programmed interruption time and the interruption time related to performance. RESULTS: The automated external defibrillators were connected for a median time of 4 minutes 47 seconds (range 31 to 1,404 seconds). CPR was performed during 45%+/-15% (mean+/-SD) of the connected time or until return of spontaneous circulation. During the automated external defibrillator connection time in the 96 patients with a shockable rhythm, CPR was performed 36%+/-20% of the time. Programmed interruption of CPR took 40%+/-15% of the automated external defibrillator connection time, and no CPR was performed related to performance during 23%+/-15% of the time. A palpable pulse was never present immediately after a shock, and return of spontaneous circulation was observed in 3 of 184 patients before arrival of the ambulance. Ultimately, return of spontaneous circulation occurred in 87 of 184 patients. CONCLUSION: First responders using automated external defibrillator voice prompts provide CPR less than half the time that the automated external defibrillator is connected to the patient. Technical improvements in automated external defibrillator rhythm analysis, more efficient resuscitation algorithms, and first-responder education could increase CPR delivery and, perhaps, improve outcome. PMID- 14520316 TI - Automated external defibrillation versus manual defibrillation for prolonged ventricular fibrillation: lethal delays of chest compressions before and after countershocks. AB - STUDY OBJECTIVE: We sought to determine whether the delays in chest compressions and defibrillation associated with an automated external defibrillator would adversely affect outcome compared with manual defibrillation in a swine model of out-of-hospital prolonged ventricular fibrillation. METHODS: After 8 minutes of untreated ventricular fibrillation, 16 swine (33+/-4 kg) were randomly assigned to automated external defibrillator defibrillation or manual defibrillation with the same biphasic truncated exponential waveform 150-J shock through the same type of pads. Defibrillation with the automated external defibrillator was performed as recommended by the manufacturer, and manual defibrillation was provided per American Heart Association Guidelines. The primary outcome measure was 24-hour survival with good neurologic outcome. Data are described as means+/ SD. RESULTS: None of 8 animals in the automated external defibrillator group survived for 24 hours, whereas 5 of 8 animals in the manual defibrillation group survived 24 hours, all with good neurologic outcome (P=.027). The time interval from simulated defibrillator arrival to first compressions was 98+/-18 seconds in the automated external defibrillator group versus 68+/-15 seconds in the manual defibrillation group. In particular, the interval from first shock to first chest compressions was 46+/-18 seconds versus 22+/-16 seconds, respectively. The mean percentage of time that chest compressions were performed in the first minute after the first countershock was 15%+/-13% versus 40%+/-15%, respectively. As a result, return of spontaneous circulation within 5 minutes of simulated defibrillator arrival occurred in only 1 of 8 animals in the automated external defibrillator group versus 6 of 8 animals in the manual defibrillation group. CONCLUSION: The longer delays in chest compressions with automated external defibrillator defibrillation versus manual defibrillation can worsen the outcome from prolonged ventricular fibrillation. PMID- 14520317 TI - The pyramid of injury: using ecodes to accurately describe the burden of injury. AB - STUDY OBJECTIVE: Although much is known about injury-related deaths from the use of external cause of injury codes (ecodes) on death certificates, the contribution of nonfatal injury is unknown, with most information based on estimates from national surveys. Some states mandate ecoding of charts for hospitalized patients, but few require ecode assignment for emergency department (ED) records. Missouri and Nebraska mandated ecoding of ED records in 1993 and 1994, respectively, allowing for a more complete description of injuries in those states. We describe fatal and nonfatal injury frequencies in Missouri and Nebraska by using ecodes, with graphic representation in the form of injury pyramids. METHODS: Ecode frequencies for 1996 to 1998 for all injury causes in Missouri and Nebraska were reported directly from their respective health departments. The ecode frequencies were grouped according to the Centers for Disease Control and Prevention's ecode matrix for presenting injury and mortality data. RESULTS: During the study period, 13,052 deaths, 131,210 hospitalizations, and 1,914,140 ED visits occurred as the result of injury. The most frequent lethal injuries were unintentional motor vehicle crashes (32.3% of total deaths), self-inflicted gunshot wound (13.2%), unintentional falls (11.3%), gunshot wound from an assault (7.7%), and unintentional poisoning (4.3%). The leading causes of injury-related hospitalization were unintentional falls (47.8% of total hospitalizations), unintentional motor vehicle crashes (15.5%), self-inflicted poisoning (6.5%), and overexertion or strenuous movements (2.4%). Of 1.9 million ED injury visits, unintentional falls accounted for 24.3%, unintentionally being struck by an object or person for 14.6%, unintentional motor vehicle crashes for 11.4%, unintentionally being cut or pierced for 10.7%, and overexertion or strenuous movements for 8.5%. CONCLUSION: Ecoding in Missouri and Nebraska provides a comprehensive data retrieval system that allows for a graphic depiction of the burden of injury derived from real patient encounters within specific geographic regions. PMID- 14520318 TI - Commentary: Pedestrian roadway fatalities: profiling the problem. PMID- 14520319 TI - Do responses to an intimate partner violence screen predict scores on a comprehensive measure of intimate partner violence in low-income black women? AB - STUDY OBJECTIVES: Screening for intimate partner violence has been advocated as an emergency department (ED) procedure. This study aimed to ascertain whether a 5 item intimate partner violence screening questionnaire could be used effectively in the ED with low-income black women to accurately predict partner abuse status. METHODS: Data were collected from 200 black women who answered in the affirmative to at least 1 item on the intimate partner violence screener questionnaire, the Universal Violence Prevention Screening Protocol. The women completed a comprehensive battery of measures, including the Index of Spouse Abuse, a commonly used and psychometrically sound measure of intimate partner violence. RESULTS: Bivariate logistic regression analyses revealed that, compared with women below the physical-intimate partner violence cut point on the Index of Spouse Abuse, women above the cut point on physical-intimate partner violence on the Index of Spouse Abuse were more likely to answer yes to Universal Violence Prevention Screening Protocol screening questions related to physical, sexual, and emotional abuse; threats to be harmed physically; and being afraid. Compared with women below the nonphysical-intimate partner violence cut point on the Index of Spouse Abuse, women above the cut point on nonphysical intimate partner violence on the Index of Spouse Abuse were more likely to answer yes to each screening question on the Universal Violence Prevention Screening Protocol. The 2 Universal Violence Prevention Screening Protocol screening items related to physical abuse best predicted the 2 Index of Spouse Abuse scales. Accurate prediction of physical and nonphysical abuse on the Index of Spouse Abuse required affirmative responses to 4 or more screening questions on the Universal Violence Prevention Screening Protocol. CONCLUSION: A brief intimate partner violence screening device in the emergency care setting can identify abused, low income, black women. The study is limited by the fact that universal screening was not conducted, the inclusion of only women who acknowledged some form of intimate partner violence, a reliance on retrospective self-reports, and the questionable generalizability of the findings to groups other than low-income black women. PMID- 14520320 TI - A decision rule for identifying children at low risk for brain injuries after blunt head trauma. AB - STUDY OBJECTIVE: Computed tomography (CT) is frequently used in evaluating children with blunt head trauma. Routine use of CT, however, has disadvantages. Therefore, we sought to derive a decision rule for identifying children at low risk for traumatic brain injuries. METHODS: We enrolled children with blunt head trauma at a pediatric trauma center in an observational cohort study between July 1998 and September 2001. We evaluated clinical predictors of traumatic brain injury on CT scan and traumatic brain injury requiring acute intervention, defined by a neurosurgical procedure, antiepileptic medications for more than 1 week, persistent neurologic deficits, or hospitalization for at least 2 nights. We performed recursive partitioning to create clinical decision rules. RESULTS: Two thousand forty-three children were enrolled, 1,271 (62%) underwent CT, 98 (7.7%; 95% confidence interval [CI] 6.3% to 9.3%) had traumatic brain injuries on CT scan, and 105 (5.1%; 95% CI 4.2% to 6.2%) had traumatic brain injuries requiring acute intervention. Abnormal mental status, clinical signs of skull fracture, history of vomiting, scalp hematoma (in children < or =2 years of age), or headache identified 97/98 (99%; 95% CI 94% to 100%) of those with traumatic brain injuries on CT scan and 105/105 (100%; 95% CI 97% to 100%) of those with traumatic brain injuries requiring acute intervention. Of the 304 (24%) children undergoing CT who had none of these predictors, only 1 (0.3%; 95% CI 0% to 1.8%) had traumatic brain injury on CT, and that patient was discharged from the ED without complications. CONCLUSION: Important factors for identifying children at low risk for traumatic brain injuries after blunt head trauma included the absence of: abnormal mental status, clinical signs of skull fracture, a history of vomiting, scalp hematoma (in children < or =2 years of age), and headache. PMID- 14520321 TI - Prediction of intracranial injury in children aged five years and older with loss of consciousness after minor head injury due to nontrivial mechanisms. AB - STUDY OBJECTIVE: Indications for computed tomography (CT) in children with minor head injury remain controversial. The objective of this preliminary study is to determine whether a clinical decision rule developed for adults could be used in children aged 5 years and older. METHODS: A prospective questionnaire was completed on all patients who were aged 5 to 17 years with major mechanisms of injury resulting in minor head injury (defined as normal Glasgow Coma Scale or modified coma scale in infants, plus normal brief neurologic examination) and loss of consciousness. The questionnaire documented 6 clinical variables: headache, emesis, intoxication, seizure, short-term memory deficits, and physical evidence of trauma above the clavicles. CT was obtained for all patients, findings were compared with the results of the questionnaires, and the sensitivity and specificity of the decision rule were determined. RESULTS: Throughout a 30-month period, 175 patients were enrolled, with a mean age of 12.8 years. Fourteen (8%) patients had intracranial injury or depressed skull fracture on CT. The presence of any of the 6 criteria was significantly associated with an abnormal CT scan result (P<.05) and was 100% (95% confidence interval 73% to 100%) sensitive for identifying patients with intracranial injury. CONCLUSION: In this preliminary study, CT use in pediatric patients with minor head injury could have been safely reduced by 23% by using a clinical decision rule previously validated in adults. PMID- 14520322 TI - Decisionmaking in pediatric minor head trauma. PMID- 14520323 TI - The death of a child in the emergency department. AB - The death of a child in the emergency department (ED) is often overwhelming to the child's community, including the health care providers involved in that child's care. Sudden death, especially of a child, induces a strong emotional response in health care providers and in the families involved. Advanced preparation by emergency staff is vital to appropriately care for the patient, the grieving family, and the ED staff. The American College of Emergency Physicians and the American Academy of Pediatrics have jointly adopted a policy statement entitled "Death of a Child in the Emergency Department Joint Statement by the American Academy of Pediatrics and the American College of Emergency Physicians." The purpose of this article is to provide the emergency physician with information related to the management of children and their families who die in the ED. The following important issues will be discussed: a family and team centered approach when a child dies, support for families and communities, communication within the child's medical home, identification of resources for use when a child dies, and critical incident stress management. PMID- 14520324 TI - Clinical policy for children younger than three years presenting to the emergency department with fever. PMID- 14520325 TI - Editorial: Clinical policy for children younger than three years presenting to the emergency department with fever. PMID- 14520327 TI - Medical error during residency: to tell or not to tell. PMID- 14520326 TI - Research without consent: current status, 2003. AB - In November 1996, regulations developed by the US Food and Drug Administration (FDA) and the Department of Health and Human Services (HHS) went into effect to allow certain emergency and resuscitation human subjects research to proceed without prospective informed consent. These new regulations brought harmonization to the requirements of the 2 federal agencies charged with research oversight and ended a moratorium that had essentially shut down resuscitation research for almost 4 years. However, the FDA's emergency exception from informed consent and the HHS's waiver of informed consent have been used infrequently. Many perceived obstacles to implementation of the regulations have been described, including the additional regulatory burden for investigators and institutional review boards, the extra expense and time required to adequately fulfill the regulatory requirements, and the reluctance of institutional review boards to allow these studies to move forward because of concerns about potential legal ramifications. Regardless of the arguments advanced, these regulations are essentially the only current regulatory options that have been provided for research without consent. This article presents a brief history of the development of the FDA's Final Rule, a summary of its requirements and its use so far, and suggestions for its implementation. Some strategies to allow the resuscitation research community to suggest fine tuning of the regulations are suggested in hopes that research requiring an exception from informed consent is allowed to proceed in a manner acceptable to regulators, is stringent in patient protection, and yet is sensitive to the practical aspects of performing resuscitation research. PMID- 14520329 TI - Age-related seasonal patterns of emergency department visits for acute asthma in an urban environment. AB - STUDY OBJECTIVE: Asthma morbidity is greater in younger patients. The reasons are not fully understood, although identifying demographic patterns of seasonality may help determine causes and potential prevention. The objective of this study is to determine the relationship between age and seasonal asthma periodicity in patients presenting to the emergency department (ED). METHODS: We conducted a retrospective study of ED visits from 1991 to 2000 in 11 municipal hospitals in New York City, with 911 receiving facilities. There were 673,141 patients who presented to the ED during the study period and had a primary diagnosis of acute asthma. RESULTS: Distinct seasonal patterns were observed, with the highest number of visits occurring in the fall and the fewest in the summer. Seasonal fluctuations of ED visits were highest in children aged 13 years or younger (coefficient of variation [CV] 37.8%; 95% confidence interval [CI] 37.5% to 38.1%), with a peak in CV occurring at approximately age 7 years (CV 43.3%; 95% CI 43.0% to 43.6%). Less variability was noted with increasing age, and the population aged 30 years and older appeared to be the least susceptible to seasonal influences (CV 11.7%; 95% CI 11.3% to 12.1%). Although the total number of asthma visits decreased by more than 30% from 1991 to 2000, the CVs for each year remained within a relatively narrow range of 24.2% to 30.5%. CONCLUSION: In an urban population, seasonal variability of asthma episodes requiring ED visits are closely linked to age, which may be important in understanding the causes of asthma and developing disease-management strategies for the prevention of asthma episodes. PMID- 14520328 TI - Financial impact on emergency physicians for nonreimbursed care for the uninsured. AB - STUDY OBJECTIVE: The financial impact on emergency physicians' reimbursement for uninsured patient care has not been previously evaluated. We conducted this study to estimate the amount of emergency physicians' nonreimbursed care for uninsured patients in Michigan. METHODS: This retrospective observational study used a convenience sample of reimbursement information from 29 hospitals. Information collected included total uninsured visits, total uninsured collections, percentage of patients uninsured, and different levels of service provided to the uninsured. Data were collected for the first quarter of 2001 and yearly data extrapolated. Expected reimbursement was estimated by using Medicare and Medicaid fee schedules. The actual amount collected was subtracted from the calculated expected amount that should be collected, and this final amount was the estimate of nonreimbursed care for the uninsured. The state estimation used American Hospital Association total emergency department (ED) visits yearly and study sample rate of uninsured patients. RESULTS: The 29 hospitals represented 1,146,280 ED visits yearly (31% of the total state ED visits). The hospitals served an average uninsured population of 11.1% (95% confidence interval [CI] 10.7% to 13.3%), with average collection per uninsured patient of $16.50 (95% CI $12.87 to $20.12). According to Medicare and Medicaid fee schedules, the total nonreimbursement per uninsured patient was $77.15 (range $73.53 to $80.78) and $61.81 (range $58.19 to $65.44), respectively. The state estimate for nonreimbursed care to the uninsured was $31,717,000 per year (range $30,227,000 to $33,208,000) according to the Medicare fee schedule and $25,408,000 per year (range $23,921,000 to $26,902,000) according to Medicaid estimations. CONCLUSION: The amount of emergency physicians' nonreimbursed care of the uninsured is substantial. PMID- 14520331 TI - Dural sinus thrombosis presenting three months postpartum. AB - Dural sinus thrombosis is a rare complication of pregnancy and the puerperium. We report a case of dural sinus thrombosis that presented as a persistent headache and then a new-onset seizure in a previously healthy 28-year-old woman 3 months postpartum. Subtle changes consistent with sinus thrombosis were present on noncontrast computed tomography, but magnetic resonance venography ultimately confirmed the diagnosis. Anticonvulsant and anticoagulant therapy were initiated and maintained, and the patient recovered completely. Follow-up genetic analysis revealed heterozygosity for the Factor II 20210A variant of the prothrombin gene mutation. PMID- 14520330 TI - Lower gastrointestinal hemorrhage from an arterioenteric fistula in a pancreatorenal transplant patient. AB - We report a case of a severe lower gastrointestinal hemorrhage caused by arterioenteric fistula formation in a pancreas transplant patient. A rare but potentially deadly complication of pancreas transplant surgery, arterioenteric fistulas should be considered in every transplant patient presenting with voluminous gastrointestinal bleeding. If diagnosed promptly, angiographic and surgical intervention can prevent serious morbidity and mortality. PMID- 14520334 TI - Hospital disaster privileging. PMID- 14520337 TI - Clozapine but not haloperidol Re-establishes normal task-activated rCBF patterns in schizophrenia within the anterior cingulate cortex. AB - Our previous work has identified that unmedicated volunteers with schizophrenia have regional cerebral blood flow (rCBF) activation patterns inappropriately related to the cognitive demand of a task in anterior cingulate cortex (ACC). Using positron emission tomography (PET) with (15)O water, we compared task induced rCBF patterns induced by haloperidol or clozapine in individuals with schizophrenia. We hypothesized that clozapine, given its superior clinical action, would tend to normalize the abnormal task-activated response in ACC more than haloperidol. Schizophrenia volunteers (SVs) (n=6) and normal volunteers (NVs) (n=12) were trained to perform a tone discrimination task with 70-80% accuracy. They were then scanned during three task conditions: (1). Rest, (2). sensory motor control (SMC) task, and (3). decision task (DEC). SVs were initially scanned after withdrawal of all psychotropic medication and again after treatment with therapeutic doses of haloperidol (n=5) and/or clozapine (n=5). rCBF values, sampled in the grown maxima of the task-activated ACC cluster, were analyzed between groups and task conditions. Task performance was similar across the unmedicated, haloperidol- and clozapine-medicated SV groups. There was a reduction in accuracy in the haloperidol SV group compared to the NVs. Group and task conditions affected rCBF in the ACC. Clozapine, but not haloperidol, reversed the abnormal ACC rCBF pattern in unmedicated SV to normal. The clozapine treated SV group showed a rCBF pattern similar to the NV group in that ACC activation was not observed during the control task but occurred during the decision condition. The pattern seen in the haloperidol-treated SV group was similar to the unmedicated SV group in that ACC activation was seen during the control task and no further activation was seen during the DEC. We report that clozapine, but not haloperidol, normalizes anterior cingulate rCBF patterns in schizophrenia during a cognitive task. Based on these preliminary data, we propose that this pattern may account for the superior therapeutic effect of clozapine and represents a surrogate marker of this action. PMID- 14520338 TI - Research funding: the problem with priorities. PMID- 14520340 TI - Organic semiconductors: An equal-opportunity conductor. PMID- 14520341 TI - Electronic paper: High-speed inks. PMID- 14520343 TI - Dilute magnetic semiconductors: Functional ferromagnets. PMID- 14520342 TI - Amorphous materials: Glasses with liquid-like surfaces. PMID- 14520344 TI - Photonic crystals: Towards rational material design. PMID- 14520345 TI - Superconductors: Clathrates join the covalent club. PMID- 14520347 TI - The never-ending 'game'. PMID- 14520348 TI - Marijuana researchers reach for pot of gold. PMID- 14520349 TI - Diabetes treatments get sweet help from nature. PMID- 14520351 TI - Bacterial battalions join war against cancer. PMID- 14520355 TI - Polio eradication scheme suffers summer setback. PMID- 14520358 TI - Tak Mak. PMID- 14520360 TI - Historical accuracy of HIV isolation. PMID- 14520361 TI - Rethinking natural-born killers. PMID- 14520362 TI - Lasker awards 2003. Foreword: It's a grand year for celebrating science. PMID- 14520363 TI - Lasker Basic Medical Research Award. The eukaryotic transcriptional machinery: complexities and mechanisms unforeseen. PMID- 14520364 TI - Lasker Clinical Medical Research Award. TNF defined as a therapeutic target for rheumatoid arthritis and other autoimmune diseases. PMID- 14520367 TI - Tumors hijack fetal enzyme, escape killer T cells. PMID- 14520368 TI - Mutants escape from killer T cells, invade population. PMID- 14520369 TI - Has gene therapy for ALS arrived? PMID- 14520370 TI - Exposing the roots of hair cell regeneration in the ear. PMID- 14520372 TI - The new immunosuppression: just kill the T cell. PMID- 14520373 TI - Beta-receptor polymorphisms: heart failure's crystal ball. PMID- 14520377 TI - Why small matters. PMID- 14520374 TI - HIV's great escape. PMID- 14520378 TI - Indian biogenerics industry emerges. PMID- 14520380 TI - First Parkinson gene therapy trial launches. PMID- 14520381 TI - EMEA struggles with need to restructure. PMID- 14520382 TI - New rules for US drug reimbursement. PMID- 14520383 TI - FDA embraces risk-management approach. PMID- 14520384 TI - Increasing number of companies found ineligible for SBIR funding. PMID- 14520385 TI - New biotech hubs may emerge as industry matures. PMID- 14520387 TI - Little science, big bucks. PMID- 14520388 TI - The 'Lilly doctrine' is viable and critical. PMID- 14520389 TI - Competing interests. PMID- 14520392 TI - Commercializing nanotechnology. PMID- 14520393 TI - Investing in nanotechnology. PMID- 14520394 TI - Ion genomics. PMID- 14520395 TI - Bacterial batteries. PMID- 14520396 TI - Breathing new life into insect-resistant plants. PMID- 14520397 TI - New paths to human ES cells? PMID- 14520398 TI - Soft landing for protein chips. PMID- 14520400 TI - The 'right' size in nanobiotechnology. AB - The biological and physical sciences share a common interest in small structures (the definition of 'small' depends on the application, but can range from 1 nm to 1 mm). A vigorous trade across the borders of these areas of science is developing around new materials and tools (largely from the physical sciences) and new phenomena (largely from the biological sciences). The physical sciences offer tools for synthesis and fabrication of devices for measuring the characteristics of cells and sub-cellular components, and of materials useful in cell and molecular biology; biology offers a window into the most sophisticated collection of functional nanostructures that exists. PMID- 14520401 TI - The potential environmental impact of engineered nanomaterials. AB - With the increased presence of nanomaterials in commercial products, a growing public debate is emerging on whether the environmental and social costs of nanotechnology outweigh its many benefits. To date, few studies have investigated the toxicological and environmental effects of direct and indirect exposure to nanomaterials and no clear guidelines exist to quantify these effects. PMID- 14520402 TI - Fabrication of novel biomaterials through molecular self-assembly. AB - Two complementary strategies can be used in the fabrication of molecular biomaterials. In the 'top-down' approach, biomaterials are generated by stripping down a complex entity into its component parts (for example, paring a virus particle down to its capsid to form a viral cage). This contrasts with the 'bottom-up' approach, in which materials are assembled molecule by molecule (and in some cases even atom by atom) to produce novel supramolecular architectures. The latter approach is likely to become an integral part of nanomaterials manufacture and requires a deep understanding of individual molecular building blocks and their structures, assembly properties and dynamic behaviors. Two key elements in molecular fabrication are chemical complementarity and structural compatibility, both of which confer the weak and noncovalent interactions that bind building blocks together during self-assembly. Using natural processes as a guide, substantial advances have been achieved at the interface of nanomaterials and biology, including the fabrication of nanofiber materials for three dimensional cell culture and tissue engineering, the assembly of peptide or protein nanotubes and helical ribbons, the creation of living microlenses, the synthesis of metal nanowires on DNA templates, the fabrication of peptide, protein and lipid scaffolds, the assembly of electronic materials by bacterial phage selection, and the use of radiofrequency to regulate molecular behaviors. PMID- 14520403 TI - Integrated nanoliter systems. AB - Microfluidic chip platforms for manipulating liquid volumes in the nanoliter range are slowly inching their way into mainstream genomic and proteomic research. The principal challenge faced by these technologies is the need for high-throughput processing of increasingly smaller volumes, with ever higher degrees of parallelization. Significant advances have been made over the past few years in addressing these needs through electrokinetic manipulation, vesicle encapsulation and mechanical valve approaches. These strategies allow levels of integration density and platform complexity that promise to make them into serious alternatives to current robotic systems. PMID- 14520404 TI - Small-scale systems for in vivo drug delivery. AB - Recent developments in the application of micro- and nanosystems for drug administration include a diverse range of new materials and methods. New approaches include the on-demand activation of molecular interactions, novel diffusion-controlled delivery devices, nanostructured 'smart' surfaces and materials, and prospects for coupling drug delivery to sensors and implants. Micro- and nanotechnologies are enabling the design of novel methods such as radio-frequency addressing of individual molecules or the suppression of immune response to a release device. Current challenges include the need to balance the small scale of the devices with the quantities of drugs that are clinically necessary, the requirement for more stable sensor platforms, and the development of methods to evaluate these new materials and devices for safety and efficacy. PMID- 14520405 TI - Electrochemical DNA sensors. AB - Electrochemistry-based sensors offer sensitivity, selectivity and low cost for the detection of selected DNA sequences or mutated genes associated with human disease. DNA-based electrochemical sensors exploit a range of different chemistries, but all take advantage of nanoscale interactions between the target in solution, the recognition layer and a solid electrode surface. Numerous approaches to electrochemical detection have been developed, including direct electrochemistry of DNA, electrochemistry at polymer-modified electrodes, electrochemistry of DNA-specific redox reporters, electrochemical amplifications with nanoparticles, and electrochemical devices based on DNA-mediated charge transport chemistry. PMID- 14520406 TI - Errors in patent application sequence listings. PMID- 14520410 TI - Converging science and technology at the nanoscale: opportunities for education and training. PMID- 14520411 TI - Widespread requirement for Hedgehog ligand stimulation in growth of digestive tract tumours. AB - Activation of the Hedgehog (Hh) signalling pathway by sporadic mutations or in familial conditions such as Gorlin's syndrome is associated with tumorigenesis in skin, the cerebellum and skeletal muscle. Here we show that a wide range of digestive tract tumours, including most of those originating in the oesophagus, stomach, biliary tract and pancreas, but not in the colon, display increased Hh pathway activity, which is suppressible by cyclopamine, a Hh pathway antagonist. Cyclopamine also suppresses cell growth in vitro and causes durable regression of xenograft tumours in vivo. Unlike in Gorlin's syndrome tumours, pathway activity and cell growth in these digestive tract tumours are driven by endogenous expression of Hh ligands, as indicated by the presence of Sonic hedgehog and Indian hedgehog transcripts, by the pathway- and growth-inhibitory activity of a Hh-neutralizing antibody, and by the dramatic growth-stimulatory activity of exogenously added Hh ligand. Our results identify a group of common lethal malignancies in which Hh pathway activity, essential for tumour growth, is activated not by mutation but by ligand expression. PMID- 14520412 TI - Molecular identification of a danger signal that alerts the immune system to dying cells. AB - In infections, microbial components provide signals that alert the immune system to danger and promote the generation of immunity. In the absence of such signals, there is often no immune response or tolerance may develop. This has led to the concept that the immune system responds only to antigens perceived to be associated with a dangerous situation such as infection. Danger signals are thought to act by stimulating dendritic cells to mature so that they can present foreign antigens and stimulate T lymphocytes. Dying mammalian cells have also been found to release danger signals of unknown identity. Here we show that uric acid is a principal endogenous danger signal released from injured cells. Uric acid stimulates dendritic cell maturation and, when co-injected with antigen in vivo, significantly enhances the generation of responses from CD8+ T cells. Eliminating uric acid in vivo inhibits the immune response to antigens associated with injured cells, but not to antigens presented by activated dendritic cells. Our findings provide a molecular link between cell injury and immunity and have important implications for vaccines, autoimmunity and inflammation. PMID- 14520413 TI - Hedgehog is an early and late mediator of pancreatic cancer tumorigenesis. AB - Hedgehog signalling--an essential pathway during embryonic pancreatic development, the misregulation of which has been implicated in several forms of cancer--may also be an important mediator in human pancreatic carcinoma. Here we report that sonic hedgehog, a secreted hedgehog ligand, is abnormally expressed in pancreatic adenocarcinoma and its precursor lesions: pancreatic intraepithelial neoplasia (PanIN). Pancreata of Pdx-Shh mice (in which Shh is misexpressed in the pancreatic endoderm) develop abnormal tubular structures, a phenocopy of human PanIN-1 and -2. Moreover, these PanIN-like lesions also contain mutations in K-ras and overexpress HER-2/neu, which are genetic mutations found early in the progression of human pancreatic cancer. Furthermore, hedgehog signalling remains active in cell lines established from primary and metastatic pancreatic adenocarcinomas. Notably, inhibition of hedgehog signalling by cyclopamine induced apoptosis and blocked proliferation in a subset of the pancreatic cancer cell lines both in vitro and in vivo. These data suggest that this pathway may have an early and critical role in the genesis of this cancer, and that maintenance of hedgehog signalling is important for aberrant proliferation and tumorigenesis. PMID- 14520414 TI - The tobacco aquaporin NtAQP1 is a membrane CO2 pore with physiological functions. AB - Aquaporins, found in virtually all living organisms, are membrane-intrinsic proteins that form water-permeable complexes. The mammalian aquaporin AQP1 has also shown CO2 permeability when expressed heterologously in Xenopus oocytes, although whether this is a biochemical curiosity or of physiological significance is a matter of debate. Here we report that, in the same expression system, a CO2 permeability comparable to that of the human AQP1 is observed for the tobacco plasma membrane aquaporin NtAQP1. NtAQP1 facilitates CO2 membrane transport in the homologous plant system at the cellular level, and has a significant function in photosynthesis and in stomatal opening. NtAQP1 overexpression heightens membrane permeability for CO2 and water, and increases leaf growth. The results indicate that NtAQP1-related CO2 permeability is of physiological importance under conditions where the CO2 gradient across a membrane is small, as is the case between the atmosphere and the inside of a plant cell. PMID- 14520415 TI - Basal body dysfunction is a likely cause of pleiotropic Bardet-Biedl syndrome. AB - Bardet-Biedl syndrome (BBS) is a genetically heterogeneous disorder characterized primarily by retinal dystrophy, obesity, polydactyly, renal malformations and learning disabilities. Although five BBS genes have been cloned, the molecular basis of this syndrome remains elusive. Here we show that BBS is probably caused by a defect at the basal body of ciliated cells. We have cloned a new BBS gene, BBS8, which encodes a protein with a prokaryotic domain, pilF, involved in pilus formation and twitching mobility. In one family, a homozygous null BBS8 mutation leads to BBS with randomization of left-right body axis symmetry, a known defect of the nodal cilium. We have also found that BBS8 localizes specifically to ciliated structures, such as the connecting cilium of the retina and columnar epithelial cells in the lung. In cells, BBS8 localizes to centrosomes and basal bodies and interacts with PCM1, a protein probably involved in ciliogenesis. Finally, we demonstrate that all available Caenorhabditis elegans BBS homologues are expressed exclusively in ciliated neurons, and contain regulatory elements for RFX, a transcription factor that modulates the expression of genes associated with ciliogenesis and intraflagellar transport. PMID- 14520416 TI - Should HIV-positive patients with lymphoma be offered stem cell transplants? AB - Advances in effective antiretroviral therapy for HIV infection have made high dose therapy and autologous stem cell transplantation possible in patients with HIV-associated lymphomas. Regimen-related toxicity is not significantly increased when antiretroviral therapy is combined with high-dose chemoradiotherapy. Durable engraftment can be seen with autologous stem cell rescue. Infectious complications can be managed with a combination of surveillance and prophylaxis. Long-term remissions of these high-risk lymphomas can be achieved with this approach. This suggests that patients with HIV-associated lymphomas should be considered for autologous transplantation in a manner similar to HIV-negative lymphoma patients. PMID- 14520417 TI - Mathematical model of peripheral blood stem cell harvest kinetics. AB - A mathematical model of peripheral blood stem cell harvests was developed, taking two new parameters R (number of recruited cells/minute) and E(f) (efficiency of collection) into consideration in addition to concentrations and collected amounts of cells. This model was tested on 241 harvest procedures in cancer patients (chemotherapy+G-CSF stimulation), donors of allogeneic PBSC, and platelet donors, using different collection procedures, with a Cobe Spectra Cell separator. The relationships between preapheresis concentrations, R, E(f) and harvested amounts of cells were complex, and different for different harvest procedures and populations of donors. However, invariably, recruitment played an important role and contributed significantly to the final harvest in all types of cells studied. For example, for the patient group, mean recruitment was 1.3 x 10(6) CD34+ cells/min and the amount of recruited cells corresponded to 65% of all collected cells. Recruitment was significantly influenced by pretreatment with chemo-therapy and/or radiotherapy. The mean recruitment values for the subgroups with limited, moderate, and extensive pretreatment were 1.65 x 10(6), 0.87 x 10(6), and 0.32 x 10(6) CD34+ cells released per minute, respectively. The finding of a quick and massive recruitment phenomenon may stimulate further research into hematopoiesis in order to maximize harvested cells. PMID- 14520418 TI - High-dose melphalan followed by radical radiotherapy for the treatment of massive plasmacytoma of the chest wall. AB - We report three cases of massive chest wall plasmacytoma, each greater than 10 cm in diameter, without evidence of overt myeloma, whom we treated with a combination of VAD chemotherapy consolidated by high-dose melphalan and autologous peripheral blood stem cell transplantation and radical radiotherapy. All three patients completed all components of their therapy without experiencing any major side effects and one patient has had a durable remission. The other two patients have had disease progression but at sites other than the original tumour. PMID- 14520419 TI - Allogeneic peripheral blood stem cell transplantation for high-risk non-Hodgkin's lymphoma. AB - A high incidence of nonrelapse mortality (NRM) has limited the use of allogeneic transplantation for poor prognosis non-Hodgkin's lymphoma (NHL). We sought to improve the outcome of allografting by utilizing Filgrastim-mobilized peripheral blood stem cells (PBSC) in combination with either standard ablative or reduced intensity conditioning. A total of 21 patients with intermediate/high-grade lymphoma and seven patients with low-grade histology were enrolled on protocols using PBSC. All patients were considered high risk for recurrence and/or NRM because of age >50 (n=16), refractory disease (n=17), failed autologous transplant (n=11) and abnormal organ function (n=2). In all, 17 patients received ablative regimens and 11 received modified conditioning including fludarabine, intravenous busulfan and ATG. Tacrolimus and mini-dose methotrexate were used for graft-versus-host-disease (GVHD) prophylaxis. Median follow-up was 38 months. Disease-free and overall survival were 57 and 58%. Seven of the 11 patients who relapsed after a previous transplant remain disease free. Four of the 10 patients with recurrent/persistent disease post transplant responded to additional therapy including withdrawal of immunosuppression+/-DLI. These results support a potent graft-versus-lymphoma effect and suggest that patients who relapse after an autologous transplant can be salvaged with an allogeneic transplant. PMID- 14520420 TI - The effect of hematopoietic growth factors on the risk of graft-vs-host disease after allogeneic hematopoietic stem cell transplantation: a meta-analysis. AB - The effect of hematopoietic growth factors on neutrophil recovery after allogeneic transplantation is well-recognized. Recent laboratory studies demonstrated that these cytokines may also modify T-cell and dendritic cell function, but whether the effect is strong enough to alter the risk of GVHD is unclear. We performed a meta-analysis to determine the effect of G-CSF or GM-CSF on the risk of nonhematopoietic outcomes after allogeneic transplantation. A search of the literature from 1986 to present yielded 18 publications in which data were provided for cohorts receiving growth factor vs either placebo or no therapy. These included nine prospective randomized studies, eight retrospective cohort studies, and one case-control study comprising a total of 1198 patients. The publication types were heterogeneous with regard to demographic and treatment characteristics, although within publications, comparative groups were generally balanced. The pooled risk ratio estimates with use of growth factor was 1.08 (95% CI 0.87-1.33, P=0.48) for grades 2-4 acute GVHD, 1.22 (95% CI 0.80-1.86, P=0.99) for grades 3-4 acute GVHD, and 1.02 (95% CI 0.82-1.26, P=0.87) for chronic GVHD. This analysis did not detect a significant change in the risk of acute or chronic GVHD after allogeneic hematopoietic stem cell transplantation when hematopoietic growth factors were used to shorten the initial period of neutropenia. PMID- 14520421 TI - Relationship between CsA trough blood concentration and severity of acute graft versus-host disease after paediatric stem cell transplantation from matched sibling or unrelated donors. AB - In order to determine optimal CsA trough blood concentrations (TBC) in the early post transplantation period, we analysed relationships between TBC and acute graft-versus-host disease (aGVHD) in paediatric SCT. A total of 94 children consecutively underwent allogeneic stem cell transplantation (SCT) from: matched sibling (MSD) (n=36), mismatched-related (MMRD) (n=3) and unrelated donors (UD) (n=55). GVHD prophylaxis usually included CsA alone or with methotrexate. Antithymocyte globulin was added in UD-SCT. TBC during the first weeks of post transplantation were estimated retrospectively by a Bayesian pharmacokinetic method and statistically associated with aGVHD. In MSD-SCT, the mean TBC during the first 2 weeks post transplantation were 42+/-10 and 90+/-7 ng/ml, respectively, in patients with grade II-IV and 0-I aGVHD (P=0.001). In SCT from UD and MMRD, TBC were 73+/-4 vs 95+/-8 ng/ml (P=0.284). For TBC >85 ng/ml, no patient developed grade II-IV aGVHD, 10 developed mild aGVHD and 30 had no aGVHD. For TBC <65 ng/ml, 7/11 patients receiving an MSD-SCT and 4/18 receiving an UD- or MMRD-SCT developed grade II-IV aGVHD. The mean TBC corresponding to each grade were: no GVHD: 101+/-10 ng/ml, mild: 77+/-11 ng/ml, moderate: 61+/-13 ng/ml, severe: 56+/-15 ng/ml (P <0.001). These results reveal a strong relationship between TBC during the early post transplantation period and the severity of aGVHD in paediatric SCT. PMID- 14520422 TI - Enhancement of cyclosporin A-induced autologous graft-versus-host disease after peripheral blood stem cell transplantation by utilizing selected CD34(+) cells. AB - Although autologous graft-versus-host disease (GVHD) can be induced by administration of cyclosporin A (CsA) after peripheral blood stem cell transplantation (PBSCT), the incidence appears to be remarkably lower compared to the incidence after bone marrow transplantation. The reduced incidence of autologous GVHD after PBSCT may be attributed to peripheral regulatory cells that are transferred with the stem cell inoculum. To determine whether transplantation of CD34-selected peripheral blood stem cells (PBSCs) leads to potentiation of autologous GVHD, five patients with malignant lymphoma were transplanted with CD34-selected PBSCs, followed by administration of CsA and interferon (IFN) gamma. Inducibility of autologous GVHD and autocytotoxic activities of peripheral blood mononuclear cells (PBMCs) after transplantation were assessed. All patients demonstrated prompt hematologic recovery. Cytotoxic activity of PBMCs against autologous lymphocytes was detectable in four of four patients analyzed during a limited period from days 14 to 34 post-transplant. An erythematous rash compatible with GVHD, confirmed by skin biopsy, developed in three of five patients. One of the three patients developed not only skin, but also gut and liver GVHD. Transplantation of the CD34-selected stem cell graft that does not accompany transfusion of regulatory cells may potentiate the inducibility of autologous GVHD by the administration of CsA and IFN-gamma. PMID- 14520423 TI - Detection of minimal residual disease by real-time PCR can be used as a surrogate marker to evaluate the graft-versus-myeloma effect after allogeneic stem cell transplantation. AB - Allogeneic stem cell transplantation (allo-SCT) is an effective and potentially curative treatment for some cases of multiple myeloma (MM). The curative efficacy of allo-SCT may be largely attributed to its immunological activity, the graft versus-myeloma (GVM) effect. To evaluate the kinetics of residual myeloma cells, we analyzed the follow-up bone marrow samples of three MM patients by means of a real-time molecular assay. We identified a consistent correlation between onset of graft-versus-host disease and disease response. These data suggest that real time molecular follow-up can be used to monitor the GVM effect and that it can be employed in the clinical setting to tailor post transplant immunomodulation. PMID- 14520424 TI - Cytomegalovirus viral load monitoring after allogeneic bone marrow transplantation in patients receiving antiviral prophylaxis. AB - Cytomegalovirus viral load measurement is a powerful new tool for monitoring of CMV disease; however, the optimal strategy for use is unknown. Weekly plasma CMV viral loads and CMV-related outcomes were monitored in 46 consecutive allogeneic bone marrow transplantation (BMT) recipients receiving standardised antiviral prophylaxis. A total of 412 CMV viral loads were quantitated in the first 100 days post transplantation with 77 positive samples (19%) in 20 patients (43%). No patient with all negative CMV viral load results developed CMV disease. Two of three patients with highly positive CMV viral loads (first positive < or =30 days post transplant, maximum viral load > or =5000 copies/ml, and > or =50% of samples positive) developed CMV disease. A total of 17 patients with positive CMV viral loads, who did not meet the criteria for highly positive, did not develop CMV disease. CMV viral load detection was higher in recipients who were CMV sero positive. In conclusion, CMV disease did not occur in the setting of a persistently negative CMV viral load. A positive CMV viral load result occurred commonly after allogeneic BMT, even in patients receiving antiviral prophylaxis. PMID- 14520425 TI - Cytomegalovirus antigenemia and outcome of patients treated with pre-emptive ganciclovir: retrospective analysis of 241 consecutive patients undergoing allogeneic hematopoietic stem cell transplantation. AB - CMV disease remains a major infectious complication after allogeneic hematopoietic stem cell transplantation (HSCT). To investigate the relationship between CMV antigenemia, treatment with ganciclovir (GCV), and outcome, we retrospectively analyzed 241 consecutive patients at risk for CMV infection who underwent allogeneic HSCT. Antigenemia-guided pre-emptive strategy with GCV was used for all patients. CMV antigenemia developed in 169 patients (70.1%), and CMV disease in 18 patients (7.5%). Multivariate analysis showed that acute GVHD (grades II-IV) was the only risk factor for developing antigenemia, and acute GVHD and advanced age for CMV disease. GCV use, as well as acute GVHD and advanced age, significantly increased the risk for bacterial and fungal infection after engraftment. Those who developed CMV antigenemia had a poorer outcome than those who did not (log-rank, P=0.0269), although the development of CMV disease worsened the outcome with only borderline significance (log-rank, P=0.0526). In conclusion, detection of antigenemia proved to be a poor prognostic factor for HSCT patients, which may be attributed to a combination of factors, including CMV disease itself, the effect of treatment, and a host status that allows for reactivation of CMV. Optimal pre-emptive strategy needs to be determined. PMID- 14520426 TI - Peri-engraftment clinical abnormalities following allogeneic hematopoietic cell transplantation: a retrospective review of 216 patients. AB - To evaluate the significance of clinical abnormalities occurring during the peri engraftment period following allogeneic hematopoietic cell transplantation (HCT), we retrospectively analyzed the data of 216 allogeneic HCT recipients. The most frequently observed peri-engraftment clinical abnormality (PECA) was noninfectious fever in 58 patients, followed by hepatic dysfunction in 39, weight gain in 22, and renal insufficiency in 11. Frequently identified predictive factors for a higher incidence of each PECA were HCT from an unrelated or mismatched donor, GVHD prophylaxis with cyclosporine alone, and rapid engraftment. Considering that donor type and GVHD prophylaxis are closely related to GVHD, these observations suggest that the development of PECAs might be associated with a graft-versus-host reaction. This hypothesis was supported by the fact that the patient group with each PECA showed a higher incidence of grades 3-4 acute or chronic extensive GVHD, with varying degrees of statistical significance. Although our data should be interpreted cautiously in view of their retrospective nature, some of the PECAs occurring after allogeneic HCT may be atypical manifestations of GVHD and may be associated with severe forms of acute or chronic GVHD. PMID- 14520427 TI - Gastric emptying after autologous haemopoietic stem-cell transplantation: a prospective trial. AB - Gastroparesis may be involved in the pathophysiology of prolonged nausea and vomiting after haemopoietic stem-cell transplantation (HSCT), but this has not been prospectively investigated. Gastric emptying (GE) was investigated in 20 patients before and 2 months after autologous HSCT. Gastrointestinal symptoms were graded prospectively and oral energy intake was recorded in parallel. Before transplant, all patients were asymptomatic and GE was within the reference range. Post transplant GE was delayed in three patients and three patients reported nausea and/or vomiting. Neither gastrointestinal symptoms nor oral energy intake post transplant discriminated between patients with or without delayed GE. Oral energy intake before transplant was lower (P=0.05), and there was a greater need for total parenteral nutrition (TPN) among patients who developed gastroparesis post transplant (P<0.05). Delayed GE after HSCT was found to be less common than had been believed from retrospective studies. Gastroparesis may be involved in some cases of prolonged nausea and vomiting after autologous HSCT but alternative explanations should be considered. Symptoms consistent with gastroparesis did not correlate with GE. Patients at risk of developing gastroparesis may be found among those with nutritional difficulties before and during the transplant course. PMID- 14520428 TI - Influence of human platelet antigen match on the success of stem cell transplantation after myeloablative conditioning. AB - Mismatches between donor and recipient for human platelet antigens (HPA) may affect the success of transplantation due to: (a) serving as minor histocompa tibility antigens and therefore render recipients at risk for graft-versus-host disease (GvHD), (b) inhibition of thrombopoiesis due to platelet antibodies. We therefore evaluated the occurrence of GvHD and need of platelet support by prospective analysis of donor-recipient pairs (n=53) for HPA-1, -2, -3, and -5 allotypes and screening for platelet antibodies prior to transplantation and in weekly intervals until day 100 after transplantation. Neither the incidence of GvHD nor the onset of thrombopoiesis, nor the CCI after platelet transfusions, nor the frequency of platelet transfusions was affected by HPA mismatches. Settings of homozygous donors vs heterozygous recipients or homozygous recipients vs heterozygous donors were not associated with any adverse effects on the outcome of the transplantation. Thus, the HPA-match does not affect the success of transplantation. PMID- 14520429 TI - Platelet chimerism by polymerase chain reaction (PCR) utilizing variable number of tandem repeats (VNTR) in allogeneic stem cell transplant in children: a new novel approach to full chimerism analysis. AB - Evaluation of chimerism following allogeneic transplantation has been performed traditionally focusing on two cellular compartments, namely lymphoid and myeloid. However, none has been described so far to evaluate platelet chimerism. In order to achieve full chimerism in all three cellular compartments, we prospectively obtained 138 samples of peripheral blood in 55 patients at different post transplant periods following allogeneic hematopoietic transplantation. Evaluation of chimerism was performed utilizing tests of variable number of tandem repeat (VNTR) and sex determination by quantitative polymerase chain reaction (PCR). Tests for platelet chimerism using platelet-rich plasma were simultaneously analyzed with samples for T-cell lymphoid and myeloid compartments. Complete donor chimerism was noted in 49 of 55 patients (89%), while the remaining six have split chimerism ranging from 34 to 98%. There is significant difference (P=0.0004) between the percentages of donor DNA in all three cellular compartments comparing the means+/-s.e.m. (myeloid 95.60+/-0.9, T-cell lymphocytes 87.6+/-1.9, and the platelets 90.8+/-1.5); however, comparison between the medians is not statistically significant. This study represents an additional step towards achieving full chimerism and the observation may help reduce the number of unnecessary platelet transfusions once chimerism is noted in that cellular compartment. PMID- 14520430 TI - Low T-cell chimerism is not followed by graft rejection after nonmyeloablative stem cell transplantation (NMSCT) with CD34-selected PBSC. AB - We investigate the feasibility of CD34-selected peripheral blood stem cell (PBSC) transplantation followed by pre-emptive CD8-depleted donor lymphocyte infusions (DLI) after a minimal conditioning regimen. Six patients with advanced hematological malignancies ineligible for a conventional myeloablative transplant (n=5) or metastatic renal cell carcinoma (n=1), and with an HLA-identical (n=4) or alternative (n=2) donor were included. The nonmyeloablative conditioning regimen consisted in 2 Gy TBI alone (n=4), 2 Gy TBI and fludarabine (RCC patient, n=1) or cyclophosphamide and fludarabine (patient who had previously received 12 Gy TBI, n=1). Post transplant immunosuppression was carried out with cyclosporin (CyA) and mycophenolate mofetil (MMF). Initial engraftment was achieved in all patients. One out of six patients (17%) experienced grade > or =2 acute GVHD only after abrupt cyclosporin discontinuation and alpha interferon therapy for life threatening tumor progression. T-cell chimerism was 23% (19-30) on day 28, 32% (10-35) on day 100, 78% (49-95) on day 180 and 99.5% (99-100) on day 365. Three out of four patients who had measurable disease before the transplant experienced a complete response. We conclude that CD34-selected NMSCT followed by CD8 depleted DLI is feasible and preserves engraftment and apparently also the graft versus-leukemia (GVL) effect. Further studies are needed to confirm this encouraging preliminary report. PMID- 14520431 TI - Bone marrow vs extramedullary relapse of acute leukemia after allogeneic hematopoietic cell transplantation: risk factors and clinical course. AB - A total of 118 consecutive adult patients with acute leukemia (78 AML, 36 ALL, and four acute mixed lineage leukemia) underwent allogeneic hematopoietic cell transplantation (HCT) after conditioning with BuCy (n=113) or a nonmyeloablative regimen of busulfan-fludarabine (n=5). After a median follow-up of 35.8 months (range, 6.4-91.0), 34 patients experienced at least one episode of leukemia relapse. Of 34 initial episodes, 14 (41%) occurred in extramedullary sites, with (n=8) or without (n=6) concomitant bone marrow involvement. The median time to relapse in the extramedullary sites was longer than that of relapse in bone marrow only (13.5 vs 6.1 months, P=0.046). Acute leukemia subtype and disease status at HCT showed an independent predictive value for overall relapse, as well as for extramedullary relapse with or without bone marrow involvement (Philadelphia chromosome positive acute leukemia vs low-risk AML, relative risk 22.68 (95% CI, 2.18-235.64); other than first CR vs first CR, relative risk 5.61 (95% CI, 1.80-17.51)), but not for bone marrow relapse. Our study suggests that there may be different pathogenetic mechanisms for bone marrow vs extramedullary relapse of acute leukemia after allogeneic HCT. The mode of relapse needs to be investigated in future reports of acute leukemia treated with allogeneic HCT. PMID- 14520432 TI - Long-term remission of APL with a second allogeneic BMT after CNS relapse following HLA-identical allogeneic BMT. AB - Second allogeneic bone marrow transplantation (BMT) for AML relapsing after an initial BMT has a poor prognosis, with a probability of a 2-y disease-free survival below 30 per cent, caused both by treatment-related mortality (TRM) and high relapse rate. While TRM is most likely due to heavy pretreatment, AML relapse after BMT may be due to resistant disease or to a poor graft-versus leukaemia (GvL) effect of the transplant. The degree of GvL may depend on individual donor/recipient immunoreactivity. In most published cases of second allogeneic BMT, both transplants were performed from the same donor. Here, we describe a patient who was first transplanted for acute promyelocytic leukaemia (APL) (AML FAB M3v) from his HLA-identical brother and received intensive immunotherapy including donor lymphocytes and IL2. He remained free from GvHD >I degrees, but developed CNS relapse. After a second BMT from another HLA-identical brother, he spontaneously developed GvHD III degrees, and has now been disease free for nearly 3 years. In this patient, long-term remission of AML relapsing after BMT was achieved by combining remission induction using an individual chemotherapy protocol with a second BMT from an alternative matched related donor and transient GvHD III degrees, which probably conferred a GVL effect. PMID- 14520433 TI - Resolution of late-onset asthma following high-dose chemotherapy. AB - We describe a patient with moderately severe (British Thorax Society Step IV/V) asthma requiring regular inhaled and oral corticosteroids to control symptoms who experienced resolution of her asthma following high-dose chemotherapy and autologous stem cell transplantation for breast cancer. As far as the authors are aware this is the first reported case. PMID- 14520434 TI - Level of minimal residual disease prior to haematopoietic stem cell transplantation predicts prognosis in paediatric patients with acute lymphoblastic leukaemia: a report of the Pre-BMT MRD Study Group. PMID- 14520435 TI - Tissue-selective therapy of cancer. AB - Instead of exploiting the differences between normal and cancer cells, seemingly unrelated anticancer modalities (from immunotherapy to hormones) exploit (a). the differences between various normal tissues and (b). tissue-specific similarities of normal and cancer cells. Although these therapies are successfully used for years to treat leukaemia and cancer, their unifying principles have never been explicitly formulated: namely, they are aimed at differentiated cells and normal tissues and target both normal and cancer cells in a tissue-specific manner. Whereas tiny differences between cancer and normal cells have yet to be successfully exploited for selective anticancer therapy, numerous tissue-specific differences (e.g. differences between melanocytes, prostate, thyroid and breast cells) provide a means to attack selectively that exact tissue that produced cancer. Despite inherent limitations, such as fostering resistance and dedifferentiation, tissue-selective therapy have enormous potentials to control cancer. PMID- 14520436 TI - A phase II trial of bryostatin-1 administered by weekly 24-hour infusion in recurrent epithelial ovarian carcinoma. AB - Bryostatin-1 is a macrocyclic lactone whose main mechanism of action is protein kinase C modulation. We investigated its activity as a weekly 24-h infusion in recurrent ovarian carcinoma. In all, 17 patients were recruited and 11 had chemotherapy-resistant disease as defined by disease progression within 4 months of last cytotoxic therapy. All were evaluable for toxicity and 14 for response. There were no disease responses and the main toxicity was myalgia. PMID- 14520438 TI - Prospective study of high-dose chemotherapy and autologous peripheral stem cell transplantation in adult patients with advanced desmoplastic small round-cell tumour. AB - A total of 10 desmoplastic small round-cell tumour patients were treated by high dose chemotherapy with stem cell support. After high-dose chemotherapy, no complete response conversion was obtained and EWS-WT1 fusion transcript detection was positive in the peripheral blood during follow-up in all patients. High-dose chemotherapy did not seem to change the results in desmoplastic small round-cell tumour. PMID- 14520437 TI - Modified de Gramont with oxaliplatin in the first-line treatment of advanced colorectal cancer. AB - We previously reported high activity for oxaliplatin and a modified de Gramont regimen (OxMdG) in a single centre study of patients with metastatic colorectal cancer. We now report results with a further 56 patients treated at 14 centres. Low rates of grade 3 and 4 toxicity were seen, with no toxic deaths. Objective response rates were CR/PR=53%; NC=34.7%; PD=12.2%. Median time to progression was 8.3 months and overall survival was 14.5 months. This regimen is more convenient than those based around the conventional de Gramont regimen but is highly active and well tolerated; it forms part of a current UK MRC phase 3 trial. PMID- 14520439 TI - CD40-targeted adenoviral GM-CSF gene transfer enhances and prolongs the maturation of human CML-derived dendritic cells upon cytokine deprivation. AB - Vaccination with autologous leukaemia-derived dendritic cells (DC) presents an adjuvant treatment option for chronic myeloid leukaemia (CML). Here, we show that high-efficiency CD40-targeted adenoviral gene transfer of GM-CSF to CML-derived DC induces long-lived maturation in the absence of exogenous cytokines and may thus ensure protracted stimulation of CML-specific T cells upon vaccination. PMID- 14520440 TI - Comparison of prognostic factors in patients in phase I trials of cytotoxic drugs vs new noncytotoxic agents. AB - The aims of this study were to identify prognostic variables for toxicity and survival in patients with cancer participating in phase I clinical trials and compare characteristics of those treated with cytotoxic chemotherapy (CT) and non cytotoxic drugs (non-CT). Data were collected from 420 (114 CT, 306 non-CT) patients enrolled in 16 phase I trials (five CT and 11 non-CT trials) in one cancer centre. Analyses of all patients were used to compare treatment groups, identify predictive variables for toxicity and to estimate prognostic factors in overall survival (OS). These were used to develop a prognostic index (PI). Multivariate analysis found those patients with better performance status, fewer sites of metastases, baseline Hb>12 g dl(-1) and WBC or LDH in the normal range had significantly better OS. Male gender, platelet count <450 x 10(9) l(-1), high WBC or treatment with a non-CT phase I agent significantly reduced the chance of grade 3/4 toxicity. Overall survival was not significantly different between the CT and non-CT groups (260 vs 192 days, P=0.47) except for those with liver metastases (228 vs 137 days, P=0.02). Overall tumour response was 4.9% (95% CI: 2.7-7.0%). The PI identified three distinct patient groups with median survival of 321, 257 and 117 days. In conclusion, entry into a phase I trial of a non-CT drug is a safe option for heavily pretreated patients with cancer. The PI generated from these data can estimate the survival probability for patients entering phase I studies. PMID- 14520441 TI - Vaccination of recurrent glioma patients with tumour lysate-pulsed dendritic cells elicits immune responses: results of a clinical phase I/II trial. AB - In this Phase I/II trial, the patient's peripheral blood dendritic cells were pulsed with an autologous tumour lysate of the glioma. Seven patients with glioblastoma and three patients with anaplastic glioma, ranging in age from 20 to 69 years, participated in this study. The mean numbers of vaccinations of tumour lysate-pulsed dendritic cells were 3.7 times intradermally close to a cervical lymph node, and 3.2 times intratumorally via an Ommaya reservoir. The percentage of CD56-positive cells in the peripheral blood lymphocytes increased after immunisation. There were two minor responses and four no-change cases evaluated by radiological findings. Dendritic cell vaccination elicited T-cell-mediated antitumour activity, as evaluated by the ELISPOT assay after vaccination in two of five tested patients. Three patients showed delayed-type hypersensitivity reactivity to the autologous tumour lysate, two of these had a minor clinical response, and two had an increased ELISPOT result. Intratumoral CD4+ and CD8+ T cell infiltration was detected in two patients who underwent reoperation after vaccination. This study demonstrated the safety and antitumour effects of autologous tumour lysate-pulsed dendritic cell therapy for patients with malignant glioma. PMID- 14520442 TI - Phase II study of liposomal doxorubicin and gemcitabine in the salvage treatment of ovarian cancer. AB - In total, 70 patients were enrolled into this phase II study, to evaluate the activity of the pegylated liposomal doxorubicin (PLD) and gemcitabine (GEM) combination in recurrent ovarian cancer patients. PLD, 30 mg m(-2), was administered on day 1 by 60' i.v. infusion, followed by GEM, 1000 mg m(-2), given by 30' i.v. on days 1 and 8; cycles were repeated every 21 days. In all, 67 patients are so far evaluable for response. Seven complete responses (10.4%, 95% CI: 3.1-17.7), 16 partial responses (23.9%, 95% CI: 13.7-34.1), 26 disease stabilisations (38.8%, 95% CI: 27.1-50.5) and 18 progressions (26.9%, 95% CI: 16.3-37.5) have been registered. Within the resistant population (n=36), the response rate was 25% (95% CI: 10.9-39.1). Within the group of platinum-sensitive patients (n=31), the response rate was 45.2% (95% CI: 27.7-62.7). A total of 443 courses are evaluable for toxicity. Grade 3-4 hematological toxicity was registered in 30 patients (42.8%), mainly represented by neutropenia (35.6%); palmar-plantar erythrodysesthesia affected 24 patients (34.2%), but it was of grade 3 in only seven of them (10%). PMID- 14520443 TI - Is Nottingham prognostic index useful after induction chemotherapy in operable breast cancer? AB - The Nottingham prognostic index (NPI), based on tumour size in breast, node involvement and Scarff-Bloom-Richardson (SBR) grading, has been shown to constitute a definitive prognostic factor of primary operable breast cancer in the adjuvant setting. We performed a retrospective study to evaluate the prognostic value of this index in 163 patients after neoadjuvant chemotherapy. Secondly, we examined the influence on survival of a revised NPI, only based on residual tumour size in breast and SBR grading in 228 patients, and consequently called breast grading index (BGI). The prognostic value of these two indices was also evaluated by replacing the SBR grade with the MSBR grade, a French modified SBR grading; the modified NPI (MNPI) and modified BGI (MBGI) were, respectively, obtained in 153 and 222 patients. At a median follow-up of 9.3 years, survival was significantly related to these four indices (P<0.001). Multivariate analysis revealed that MBGI was the only one which retained a prognostic influence on disease-free survival (P<0.02). In conclusion, the 'amount' of residual tumour in breast and/or nodes, as defined by NPI and revised indices, confers a determinant prognosis after neoadjuvant chemotherapy, inviting an alternative postsurgical treatment for a subgroup of patients with a decreased survival. PMID- 14520445 TI - Seasonal differences in the onset of the EBV-positive and -negative forms of paediatric Hodgkin's lymphoma. AB - In this study, we have shown that there are seasonal differences in the onset of the (Epstein-Barr virus) EBV-positive and -negative forms of paediatric Hodgkin's lymphoma (HL). This suggests aetiological differences between the two forms of this disease. EBV-positive HL might be a rare consequence of primary EBV infection. PMID- 14520444 TI - First-line gemcitabine with cisplatin or epirubicin in advanced non-small-cell lung cancer: a phase III trial. AB - The purpose of our study was to compare progression-free survival and quality of life (QOL) after cisplatin-gemcitabine (CG) or epirubicin-gemcitabine (EG) in chemotherapy-naive patients with unresectable non-small-cell lung cancer. Patients (n=240) were randomised to receive gemcitabine 1125 mg x m(-2) (days 1 and 8) plus either cisplatin 80 mg x m(-2) (day 2) or epirubicin 100 mg x m(-2) (day 1) every 3 weeks for a maximum of five cycles. Eligible patients had normal organ functions and Eastern Cooperative Oncology Group performance status or=30) compared with normal weighted was 1.09 (95% CI: 1.04-1.15). However, the RR at age 50-59 years was 1.58 (95% CI: 1.29-1.94) in men being obese at about age 45 years compared with normal weighted men. The tallest men had an RR of 1.72 (95% CI: 1.46-2.04) compared with the shortest men. The overall effect of BMI on the incidence of prostate cancer was modest. The larger effect found in men aged 50-59 years might partly explain the previous inconsistent findings. PMID- 14520454 TI - Blood pressure and site-specific cancer mortality: evidence from the original Whitehall study. AB - Studies relating blood pressure to cancer risk have some shortcomings and have revealed inconsistent findings. In 17498 middle-aged London-based government employees we related systolic and diastolic blood pressure recorded at baseline examination (1967-1970) to the risk of cancer mortality risk at 13 anatomical sites 25 years later. Following adjustment for potential confounding and mediating factors, inverse associations between blood pressure and mortality due to leukaemia and cancer of the pancreas (diastolic only) were seen. Blood pressure was also positively related to cancer of the liver and rectum (diastolic only). The statistically significant blood pressure-cancer associations seen in this large-scale prospective investigation offering high power were scarce and of sufficiently small magnitude as to be attributable to chance or confounding. PMID- 14520455 TI - Seroprevalence of human papillomavirus-16, -18, -31, and -45 in a population based cohort of 10000 women in Costa Rica. AB - Human papillomavirus (HPV) seroprevalence and determinants of seropositivity were assessed in a 10049-woman population-based cohort in Guanacaste, Costa Rica. Serologic responses based on VLP-based ELISA were obtained from the plasma collected at study enrollment in 1993/1994 for HPV-16 (n=9949), HPV-18 (n=9928), HPV-31 (n=9932), and HPV-45 (n=3019). Seropositivity was defined as five standard deviations above the mean optical density obtained for studied virgins (n=573). HPV-16, -18, -31, and -45 seroprevalence was 15, 15, 16, and 11%, respectively. Of women DNA-positive for HPV-16, -18, -31, or -45, seropositivity was 45, 34, 51, and 28%, respectively. Peak HPV seroprevalence occurred a decade after DNA prevalence; lifetime number of sexual partners was the key determinant of seropositivity independent of DNA status and age. DNA- and sero-positive women showed the highest risk for concurrent CIN3/cancer, followed by DNA-positive, sero-negative women. PMID- 14520457 TI - Combining cohort and period methods for retrospective time trend analyses of long term cancer patient survival rates. AB - Assessing trends in long-term cancer patient survival is an essential component of monitoring progress against cancer by cancer registries. Traditional assessment of long-term survival ('cohort analysis') is very useful to disclose trends in long-term survival rates of patients diagnosed many years ago, but it does not allow the disclosure of recent trends in long-term survival rates. The latter can be achieved by an alternative method of survival analysis ('period analysis'), which has been proposed a few years ago. On the other hand, unlike cohort analysis, period analysis does not provide estimates of long-term survival rates for patients diagnosed in the early years after initiation of cancer registration. In this paper, a method of retrospective analysis of time trends in long-term survival rates is introduced, which combines the advantages of both cohort and period analysis ('mixed analysis'). This method thereby allows for a comprehensive monitoring of trends in long-term survival over an extended time span from the earliest to the most recent years of cancer registration. The use of the method is illustrated for retrospective time trend analyses of long-term survival of cancer patients in the United States with the 1973-1999 database of the Surveillance, Epidemiology, and End Results Program of the National Cancer Institute. PMID- 14520456 TI - Flavonoid intake and breast cancer risk: a case--control study in Greece. AB - Flavonoids have been investigated for possible inverse associations with various chronic degenerative diseases, but there are no epidemiologic data concerning a possible association between several of the main flavonoid categories and breast cancer risk. We have applied recently published data on the flavonoid content of several foods and beverages on dietary information collected in the context of a large case-control study of 820 women with breast cancer and 1548 control women, conducted in Greece. We found a strong, statistically significant inverse association of flavone intake with breast cancer. The odds ratio for an increment equal to one standard deviation of daily flavone intake (i.e. 0.5 mg day(-1)) was 0.87, with 95% confidence interval 0.77-0.97. The association persisted after controlling for fruit and vegetable consumption, or for other flavonoid intake. This inverse association is compatible with and may explain the reported inverse association of breast cancer with consumption of vegetables, particularly leafy vegetables. After controlling for dietary confounding, there was no association of breast cancer risk with flavanones, flavan-3-ols, flavonols, anthocyanidins or isoflavones. PMID- 14520458 TI - Membranous location of EGFR immunostaining is associated with good prognosis in renal cell carcinoma. AB - Epidermal growth factor receptor (EGFR) is a key factor in tumorigenesis. The association between EGFR expression and prognosis in renal cell carcinoma (RCC) is not clear. In our study of 134 RCCs, the cellular location of immunostaining was evaluated and patients with EGFR-positive tumours with prominent membranous staining had a good prognosis. Their overall survival was significantly longer (P=0.004) than that of patients with either EGFR-negative tumours or with mainly cytoplasmic staining. However, further studies on the different EGFR expression patterns in RCC are needed to clarify their role in the progression of the disease. PMID- 14520459 TI - Matrix metalloproteinase-2 (MMP-2) is associated with survival in breast carcinoma. AB - Adjuvant therapy is one of the major advances in the treatment of breast carcinoma patients - but do all patients need it? New predictive markers, which are able to save breast carcinoma patients from the most toxic adjuvant therapies, are still needed. The expression of matrix metalloproteinases (MMP-2) has been previously linked to invasiveness of carcinoma cells. In this study, we explored the role of MMP-2 as a prognostic factor in breast carcinoma in a large series to be able to show the favourable effect of MMP-2 negativity in poor prognosis subgroup of hormone receptor-negative patients. The MMP-2 immunoreactive protein was evaluated from primary adenocarcinoma of the breast in 453 cases by using a specific monoclonal antibody in immunohistochemical stainings. The MMP-2 protein found in breast carcinoma tumour cells was here shown to be associated with a shortened recurrence-free survival or relative overall survival (P=0.03). It was shown here that MMP-2 negativity is significantly linked to favourable prognosis in patients considered to be at risk due to their hormone receptor negativity. In the patient group presenting with a progesterone receptor-negative tumour, the survival rate of the MMP-2-positive cases was 58% while it was 95% in MMP-2-negative cases after 10 years of follow up (P=0.005). The present data shows for the first time that MMP-2 negativity could serve as a marker for favourable prognosis in breast carcinoma patients with a hormone receptor-negative tumour usually associated with high risk. MMP-2 is also shown to correlate to shortened survival independent of major prognostic indicators in patients with primary breast carcinoma. PMID- 14520460 TI - FGFR3IIIS: a novel soluble FGFR3 spliced variant that modulates growth is frequently expressed in tumour cells. AB - Fibroblast growth factor receptor 3 (FGFR3) is one of four high-affinity tyrosine kinase receptors for the FGF family of ligands, frequently associated with growth arrest and induction of differentiation. The extracellular immunoglobulin (IgG) like domains II and III are responsible for ligand binding; alternative usage of exons IIIb and IIIc of the Ig-like domain III determining the ligand-binding specificity of the receptor. By reverse transcriptase polymerase chain reaction (RT-PCR) a novel FGFR3IIIc variant FGFR3IIIS, expressed in a high proportion of tumours and tumour cell lines but rarely in normal tissues, has been identified. Unlike recently described nonsense transcripts of FGFR3, the coding region of FGFR3IIIS remains in-frame producing a novel protein. The protein product is coexpressed with FGFR3IIIc in the membrane and soluble cell fractions; expression in the soluble fraction is decreased after exposure to bFGF but not aFGF. Knockout of FGFR3IIIS using antisense has a growth-inhibitory effect in vitro, suggesting a dominant-negative function for FGFR3IIIS inhibiting FGFR3-induced growth arrest. In summary, alternative splicing of the FGFR3 Ig-domain III represents a mechanism for the generation of receptor diversity. FGFR3IIIS may regulate FGF and FGFR trafficking and function, possibly contributing to the development of a malignant phenotype. PMID- 14520461 TI - LRIG1 and epidermal growth factor receptor in renal cell carcinoma: a quantitative RT--PCR and immunohistochemical analysis. AB - In all, 31 renal cell carcinomas (RCCs) were examined for expression of the potential tumour suppressor LRIG1 (formerly Lig-1) and the epidermal growth factor receptor (EGFR). Eight matched samples of uninvolved kidney cortex were also evaluated. Gene expression was examined by quantitative real-time RT-PCR. In the eight matched sample pairs (uninvolved kidney cortex and tumour), protein expression was examined by immunohistochemistry. Conventional (clear cell) tumours showed an expected upregulation of EGFR. LRIG1 expression was generally downregulated in conventional and papillary RCC but not in chromophobic RCC. The ratio between EGFR and LRIG1 was more than 2.5-fold higher in the eight tumours compared with matched uninvolved kidney cortex and was at least two-fold higher than the mean normal ratio in 21 of 31 samples analysed. The observed downregulation of LRIG1 and increased EGFR/LRIG1 ratios are consistent with LRIG1 being a suppressor of oncogenesis in RCC by counteracting the tumour-promoting properties of EGFR. Further studies are justified to elucidate the explicit role of LRIG1 in the oncogenesis of RCC. PMID- 14520462 TI - GLUT1 and CAIX as intrinsic markers of hypoxia in bladder cancer: relationship with vascularity and proliferation as predictors of outcome of ARCON. AB - Glucose transporter-1 protein (GLUT1) and carbonic anhydrase IX (CAIX) are regulated by hypoxia inducible factor-1 (HIF-1) and have been studied as putative intrinsic cellular markers for hypoxia. This study directly compares CAIX and GLUT1 with pimonidazole binding in a prospective series of bladder cancer patients and also studies the prognostic significance of the markers, in combination with vascularity and proliferation, in a retrospective series of bladder cancer patients treated in a phase II trial of radical radiotherapy with carbogen and nicotinamide (ARCON). A total of 21 patients with a diagnosis of transitional cell carcinoma of the bladder received 0.5 g m(-2) pimonidazole. Serial tumour sections were stained for pimonidazole, GLUT1 and CAIX and compared. Tissue sections obtained from a series of 64 patients previously treated for invasive bladder cancer using ARCON were stained for GLUT1 and CAIX together with Ki-67 and CD31/34. There was a good geographical colocalisation of both intrinsic markers with pimonidazole and a highly significant agreement in individual patients; correlation coefficients were 0.82 (P=0.0001) for GLUT1 and 0.74 (P<0.0001) for CAIX. In both series of patients, the intrinsic hypoxia markers were highly correlated with each other and a correlation with proliferation was also evident in the retrospective study. In univariate and multivariate analyses, GLUT1 and CAIX were independent predictors for overall and cause specific survival. The hypoxia markers did not predict for local control or metastases-free survival although higher Ki-67 indices showed a trend towards local failure. The data suggest that both hypoxia modification and accelerated treatment may be valid treatment options in bladder cancer. PMID- 14520463 TI - Wnt-signalling pathway in ovarian epithelial tumours: increased expression of beta-catenin and GSK3beta. AB - Beta-catenin is involved in both cell-cell adhesion and in transcriptional regulation by the Wingless/Wnt signalling pathway. Alterations of components of this pathway have been suggested to play a central role in tumorigenesis. The present study investigated, by immunohistochemistry and immunoblotting, the protein expression and localisation of beta-catenin, adenomatous polyposis coli (APC), glycogen synthase kinase 3beta (GSK3beta) and lymphocyte enhancer factor-1 (Lef-1) in normal human ovaries and in epithelial ovarian tumours in vivo and in vitro. Immortalised human ovarian surface epithelium and ovarian cancer cell cells (OVCAR-3) expressed beta-catenin, APC, GSK3beta and Lef-1. Nuclear staining of beta-catenin and Lef-1 were demonstrated only in OVCAR-3 cells. There were significant increases of beta-catenin and GSK3beta, while APC was reduced in ovarian cancer compared to the normal ovary. Beta-catenin and Lef-1 were coimmunoprecipitated in ovarian tumours, but not in the normal ovary. Nuclear localisation of beta-catenin or Lef-1 could not be demonstrated in the normal ovary or in the ovarian tumours. The absence of nuclear localisation of beta catenin could be due to an increased binding to the cadherin-alpha-catenin cell adhesion complex. In fact, we have earlier reported an increased expression of E cadherin in ovarian adenocarcinomas. In summary, this study demonstrates an increase in the expression of components of the Wingless/Wnt pathway in malignant ovarian tumours. The increase suggests a role for this signalling pathway in cell transformation and in tumour progression. However, it remains to be demonstrated whether it is an increased participation of beta-catenin in transcriptional regulation, or in the stabilisation of cellular integrity, or both, that is the crucial event in ovarian tumorigenesis. PMID- 14520464 TI - HER2/neu overexpression in the development of muscle-invasive transitional cell carcinoma of the bladder. AB - The mortality from transitional cell carcinoma (TCC) of the urinary bladder increases significantly with the progression of superficial or locally invasive disease (pTa/pT1) to detrusor muscle-invasive disease (pT2+). The most common prognostic markers in clinical use are tumour stage and grade, which are subject to considerable intra- and interobserver variation. Polysomy 17 and HER2/neu gene amplification and protein overexpression have been associated with more advanced disease. Standardised techniques of fluorescence in situ hybridisation and immunohistochemistry, which are currently applied to other cancers with a view to offering anti-HER2/neu therapies, were applied to tumour pairs comprising pre- and postinvasive disease from 25 patients undergoing treatment for bladder cancer. In the preinvasive tumours, increased HER2/neu copy number was observed in 76% of cases and increased chromosome 17 copy number in 88% of cases, and in the postinvasive group these values were 92 and 96%, respectively (not significantly different P=0.09 and 0.07, respectively). HER2 gene amplification rates were 8% in both groups. Protein overexpression rates were 76 and 52%, respectively, in the pre- and postinvasive groups (P=0.06). These results suggest that HER2/neu abnormalities occur prior to and persist with the onset of muscle invasive disease. Gene amplification is uncommon and other molecular mechanisms must account for the high rates of protein overexpression. Anti-HER2/neu therapy might be of use in the treatment of TCC. PMID- 14520465 TI - Diagnosis of axillary nodal metastases by ultrasound-guided core biopsy in primary operable breast cancer. AB - The purpose of this study was to examine the use of ultrasound (US)-guided core biopsy of axillary nodes in patients with operable breast cancer. The ipsilateral axillae of 187 patients with suspected primary operable breast cancer were scanned. Nodes were classified based on their shape and cortical morphology. Abnormal nodes underwent US-guided core biopsy/fine needle aspiration (FNA), and the results correlated with subsequent axillary surgery. The nodes were identified on US in 103 of 166 axillae of patients with confirmed invasive carcinoma. In total, 54 (52%) met the criteria for biopsy: 48 core biopsies (26 malignant, 20 benign node, two normal) and six FNA were performed. On subsequent definitive histological examination, 64 of 166 (39%) had axillary metastases. Of the 64 patients with involved nodes at surgery, preoperative US identified nodes in 46 patients (72%), of which 35 (55%) met the criteria for biopsy and 27 (42%) of these were diagnosed preoperatively by US-guided biopsy. In conclusion, US can identify abnormal nodes in patients presenting with primary operable breast cancer. In all, 65% of these nodes are malignant and this can often be confirmed with US-guided core biopsy. PMID- 14520466 TI - Detection of p53 mutations in precancerous gastric tissue. AB - Intestinal-type gastric cancer is preceded by gastritis and intestinal metaplasia. There is uncertainty regarding the stage at which genetic alterations in the p53 gene occur. Reactive oxygen species (ROS) may participate in the production of mutations and the inactivation of p53 is due to infection by the bacterium Helicobacter pylori. We have investigated whether alterations of the p53 gene can be detected in gastritis and intestinal metaplasia using the restriction site mutation assay. We also assessed the potential contribution of ROS to p53 inactivation using electron spin resonance spectroscopy (ESR) and correlated with the presence of H. pylori. In all, 35% of the gastritis samples and 45% of the intestinal metaplasia samples were found to contain mutations in exons 5-8 of the p53 gene. Electron spin resonance spectroscopy analysis showed a significant increase in free radical levels in gastritis samples compared with normal, intestinal metaplasia and cancer samples, suggesting that free radicals present in gastritis may contribute to p53 mutations. There was no significant difference in free radical levels between the H. pylori-positive and -negative groups. However, a small subpopulation of the H. pylori-negative patients had much higher levels of free radicals. This suggests a more prominent role for other factors in ROS production. PMID- 14520467 TI - Involvement of nucleophosmin/B23 in TPA-induced megakaryocytic differentiation of K562 cells. AB - Human myelogenous leukaemia K562 cells were induced to undergo megakaryocytic differentiation by treatment with phorbol ester 12-O-tetradecanoylphorbol-13 acetate (TPA) (20 nM, 24-72 h). The steady-state level of nucleophosmin/B23 mRNA decreased during the TPA-induced differentiation. There was also decrease in the level of cellular nucleophosmin/B23 protein and appearance of its degraded product (25 kDa) during the TPA-induced differentiation. Furthermore, K562/B23 (wild type), K562/D1 (delta280-294) and K562/D2 (delta263-294) cells were less, while K562/D3 (delta244-294) cells were more responsive to TPA-induced differentiation as compared to K562/vector or parental K562 cells. Activation of the ERK/MAPK was observed in parental K562 cells upon TPA treatment (5 nM, 5-30 min). As compared to K562/vector cells, less activation of ERK/MAPK was observed in K562/D2 cells, while ERK/MAPK was highly activated in K562/D3 cells upon TPA treatment. Our results indicate that nucleophosmin/B23 plays an important role in TPA-induced differentiation of K562 cells and the amino acids 244-294 at C terminal of nucleophosmin/B23 could be an important site for regulation of cellular response to differentiation. PMID- 14520468 TI - In vitro selectivity, in vivo biodistribution and tumour uptake of annexin V radiolabelled with a positron emitting radioisotope. AB - The availability of a noninvasive method to detect and quantify apoptosis in tumours will enable tumour response to several cancer therapies to be assessed. We have synthesised two radiotracers, annexin V and the N-succinimidyl-3 iodobenzoic acid (SIB) derivative of annexin V, labelled with radio-iodine ((124)I and (125)I) and provided proof of the concept by assessing specific binding and biodistribution of these probes to apoptotic cells and tumours. We have also assessed the tumour uptake of [(124)I]annexin V in a mouse model of apoptosis. RIF-1 cells induced to undergo apoptosis in vitro showed a drug concentration-dependent increased binding of [(125)I]annexin V and [(125)I]SIB annexin V. In the same model system, there was an increase in terminal deoxynucleotidyl transferase-mediated nick end labelling (TUNEL)-positive cells and a decrease in clonogenic survival. Radiotracer binding was completely inhibited by preincubation with unlabelled annexin V. In RIF-1 tumour-bearing mice, rapid distribution of [(125)I]SIB-annexin V-derived radioactivity to kidneys was observed and the radiotracer accumulated in urine. The binding of [(125)I]SIB-annexin V to RIF-1 tumours increased by 2.3-fold at 48 h after a single intraperitoneal injection of 5-fluorouracil (165 mg kg(-1) body weight), compared to a 4.4-fold increase in TUNEL-positive cells measured by immunostaining. Positron emission tomography images with both radiotracers demonstrated intense localisation in the kidneys and bladder. Unlike [(124)I]SIB annexin V, [(124)I]annexin V also showed localisation in the thyroid region presumably due to deiodination of the radiolabel. [(124)I]SIB-annexin V is an attractive candidate for in vivo imaging of apoptosis by PET. PMID- 14520469 TI - Microvascular mechanisms by which the combretastatin A-4 derivative AC7700 (AVE8062) induces tumour blood flow stasis. AB - We previously reported that a novel combretastatin A-4 derivative, AC7700, has remarkable antitumour effects because of an irreversible stasis of tumour blood flow (TBF) and subsequent loss of nutrient supply to tumour tissue. Since early 2002, under the new designation AVE8062, AC7700 has undergone clinical trials in Europe and the US. Questions remain, however, concerning how AC7700 blocks TBF and why the TBF stasis does not recover. In this study, using a rat tumour LY80, a variant of Yoshida sarcoma, we examined whether TBF cessation after AC7700 administration is due to a direct action of the agent on tumour blood vessels. We constructed electrodes that can drop a small quantity of the drug solution directly at the site of blood flow measurement and inserted them subcutaneously and into the tumour. We compared the blood flow responses of normal vessels and tumour vessels after administration of 10-microl doses of various concentrations (0.2, 1, 10, and 50 mg ml(-1)) of the AC7700 solution. In addition, we assessed TBF stasis after i.v. and intra-arterial 10 mg x kg(-1) AC7700 administration in an LY80-induced kidney tumour. To determine why the TBF stasis is irreversible, we observed AC7700-induced changes in host arterioles and the tumour vascular network of the Sato lung carcinoma using a vital microscopic rat transparent chamber. Since an increase in tumour interstitial fluid pressure brings about a decrease in TBF, we also measured 10 mg x kg(-1) AC7700-induced changes in this pressure. The sensitivity of the blood flow response after intratumoral application of AC7700 was markedly higher in normal vessels relative to tumour vessels. Intra-arterial administration of AC7700 did not have stronger effects on TBF stasis than did i.v. administration. Intravital microscopy showed that AC7700 induced a powerful and long-lasting constriction of host arterioles, so that complete stasis of blood flow occurred in downstream vessels, which supplied blood to tumours. Owing to this stasis, the lumens of numerous tumour vessels narrowed or completely disappeared, and numerous erythrocytes stagnated in drainage vessels of the tumour vascular network. Haemolysis of these erythrocytes occurred after 2-3 h, resulting in complete thrombosis. There was no indication of reperfusion in vessels showing haemolysis. This haemolysis is thought to be the main cause for the irreversibility of TBF stasis. Since the tumour interstitial fluid pressure decreased after i.v. AC7700 administration, the possibility of stasis of TBF being caused by tumour vascular compression was excluded. All these results strongly suggest that the main target of AC7700 is host arterioles and that the stasis of TBF induced by AC7700 is not triggered by a direct action of the drug on tumour vessels. PMID- 14520470 TI - Fas ligand mediates immune privilege and not inflammation in human colon cancer, irrespective of TGF-beta expression. AB - Many cancers express Fas ligand (FasL/CD95L) in vivo, and can kill lymphoid cells by Fas-mediated apoptosis in vitro. However, overexpression of recombinant FasL in murine tumour allografts revealed a potential antitumour effect of FasL, via recruitment of neutrophils. Transforming growth factor-beta1 (TGF-beta1) could inhibit these neutrophil-stimulatory effects of FasL. In the present study, we sought to determine directly whether FasL contributes to immune privilege or tumour rejection in human colon cancers in vivo, and whether TGF-beta1 regulates FasL function. Serial tumour sections were immunostained for FasL and TGF-beta1. Neutrophils and tumour infiltrating lymphocytes (TILs) were detected by immunohistochemistry for lactoferrin and CD45, respectively. Apoptotic TIL were identified by dual staining for TUNEL/CD45. FasL expression by nests of tumour cells was associated with a mean four-fold depletion of TILs (range 1.8-33-fold, n=16, P<0.001), together with a two-fold increase in TIL apoptosis (range 1.6-2.5 fold, n=14, P<0.001), relative to FasL-negative nests within the same tumours. The overall level of neutrophils present in all tumours examined was low (mean 0.3%, n=16), with FasL expression by tumour nests associated with a mean two-fold decrease in neutrophils, irrespective of TGF-beta1 expression. Together, our results suggest that tumour-expressed FasL is inhibitory rather than stimulatory towards antitumour immune responses. PMID- 14520471 TI - Bcl-x(L) antisense oligonucleotides radiosensitise colon cancer cells. AB - Advanced colon cancer is a malignancy with poor response to various treatment modalities including ionising radiation (IR) and chemotherapy. Both IR and chemotherapeutic agents have been shown to act by inducing apoptosis, a type of cell death antagonised by the Bcl-x(L) gene product. Since approximately 60% of human colon cancers express Bcl-x(L), it was the aim of this study to explore the potential of Bcl-x(L) antisense oligonucleotides as a novel radiosensitisation strategy. Caco-2 colon cancer cells were treated with Bcl-x(L) antisense oligonucleotides in combination with IR or cisplatin, and Bcl-x(L) protein expression, apoptosis, cell viability and clonogenic survival were examined. Bcl x(L) antisense oligonucleotide specifically reduced the Bcl-x(L) protein level by almost 50% in Caco-2 cells. The decreased threshold for the induction of apoptosis resulted in a 300% increase of apoptosis after IR or cisplatin treatment and led to a 60% reduction of cell proliferation beyond response rates achieved with IR. These data suggest that Bcl-x(L) is an important factor contributing to the treatment resistance of human colon cancer. Specific reduction of Bcl-x(L) protein levels by antisense oligonucleotides qualifies as a promising therapeutic strategy for colon cancer that may help overcome resistance and improve clinical outcome in this malignancy. PMID- 14520472 TI - Increased NF-kappaB DNA binding but not transcriptional activity during apoptosis induced by the COX-2-selective inhibitor NS-398 in colorectal carcinoma cells. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit colorectal neoplasia, an effect that is associated with their ability to induce apoptosis. Although NSAIDs have been reported to inhibit NF-kappaB, more recent studies show activation of NF-kappaB by NSAIDs. NF-kappaB commonly shows antiapoptotic activity and is implicated in the therapeutic resistance of cancer cells. The effects of highly COX-2-selective NSAIDs such as NS-398 on NF-kappaB in colorectal tumour cells have not been reported. Therefore, we addressed whether NF-kappaB has a role in NS-398-induced apoptosis of colorectal cancer cells. Treatment of HT-29 colorectal carcinoma cells with doses of NS-398 (50-75 microM) known to induce apoptosis had no effect on NF-kappaB for up to 48 h. However after 72 and 96 h NF kappaB DNA-binding activity was increased by NS-398, in parallel with apoptosis induction. NS-398-treated HT-29 cells showed increased p50 homodimer binding and an induction of p50/p65 heterodimers, as demonstrated by supershift assay. However, although NS-398 increased NF-kappaB DNA binding it did not increase NF kappaB-dependent reporter activity and inhibition of NF-kappaB DNA binding did not enhance NS-398-induced apoptosis. This indicates that NF-kappaB activated by NS-398 is transcriptionally inactive and is an encouraging result for the use of COX-2-selective NSAIDs not only in chemoprevention but also as novel therapies for colon cancer. PMID- 14520475 TI - Uptake of meta-iodobenzylguanidine in neuroendocrine tumours is mediated by vesicular monoamine transporters. AB - The radio-iodinated noradrenaline analogue meta-iodobenzylguanidine (MIBG) can be used for scintigraphy and radiation therapy of neuroendocrine (NE). The aim of the present study was to study the importance of vesicular monoamine transporters (VMATs) for the uptake of (123)I-MIBG in NE tumours. In nude mice, bearing the human transplantable midgut carcinoid GOT1, all organs and xenografted tumours accumulated (123)I after i.v. injection of (123)I-MIBG. A high concentration of (123)I was maintained in GOT1 tumours and adrenals, which expressed VMATs, but rapidly decreased in all other tissues. In the VMAT-expressing NE tumour cell lines GOT1 and BON and in VMAT-expressing primary NE tumour cell cultures (carcinoids, n=4 and pheochromocytomas, n=4), reserpine significantly reduced the uptake of (123)I-MIBG. The membrane pump inhibitor clomipramine had no effect on the uptake of (123)I-MIBG in GOT1 and BON cells, but inhibited the uptake in one out of four primary carcinoid cell cultures and three out of four primary pheochromocytoma cell cultures. In conclusion, VMATs and secretory granules are of importance for the uptake and retention of (123)I-MIBG in NE tumours. Information about the type and degree of expression of VMATs in NE tumours may be helpful in future to select patients suitable for radiation therapy with radio iodinated MIBG. PMID- 14520480 TI - CADASIL: new cases and new questions. AB - We described the first two unrelated Polish families with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). In the morphological examination with light microscopy, two kinds of changes were observed: (1). panarteritis nodosa-like changes with eosinophilic fibrinoid necrosis of the vessel wall and perivascular inflammatory infiltrates and (2). basophilic granular material in the tunica media characteristic of CADASIL. At electron microscopy, we found deposits of granular osmophilic material (GOM) within the wall of arteries, veins and capillary vessels. Our findings imply two questions requiring further investigation: Why in the genetically determined vascular disorder are the features of systemic inflammatory vascular disease present? Why in capillary walls deprived of smooth muscle cells are deposits of GOM present? PMID- 14520474 TI - Role of glucose and ketone bodies in the metabolic control of experimental brain cancer. AB - Brain tumours lack metabolic versatility and are dependent largely on glucose for energy. This contrasts with normal brain tissue that can derive energy from both glucose and ketone bodies. We examined for the first time the potential efficacy of dietary therapies that reduce plasma glucose and elevate ketone bodies in the CT-2A syngeneic malignant mouse astrocytoma. C57BL/6J mice were fed either a standard diet unrestricted (SD-UR), a ketogenic diet unrestricted (KD-UR), the SD restricted to 40% (SD-R), or the KD restricted to 40% of the control standard diet (KD-R). Body weights, tumour weights, plasma glucose, beta-hydroxybutyrate (beta-OHB), and insulin-like growth factor 1 (IGF-1) were measured 13 days after tumour implantation. CT-2A growth was rapid in both the SD-UR and KD-UR groups, but was significantly reduced in both the SD-R and KD-R groups by about 80%. The results indicate that plasma glucose predicts CT-2A growth and that growth is dependent more on the amount than on the origin of dietary calories. Also, restriction of either diet significantly reduced the plasma levels of IGF-1, a biomarker for angiogenesis and tumour progression. Owing to a dependence on plasma glucose, IGF-1 was also predictive of CT-2A growth. Ketone bodies are proposed to reduce stromal inflammatory activities, while providing normal brain cells with a nonglycolytic high-energy substrate. Our results in a mouse astrocytoma suggest that malignant brain tumours are potentially manageable with dietary therapies that reduce glucose and elevate ketone bodies. PMID- 14520473 TI - CD26/dipeptidyl peptidase IV enhances expression of topoisomerase II alpha and sensitivity to apoptosis induced by topoisomerase II inhibitors. AB - CD26/dipeptidyl peptidase IV (DPPIV) is a cell surface-bound ectopeptidase with important roles in T-cell activation and tumour biology. We now report that CD26/DPPIV enhances sensitivity to apoptosis induced by the antineoplastic agents doxorubicin and etoposide. In particular, CD26/DPPIV presence is associated with increased susceptibility to the mitochondrial pathway of apoptosis, documented by enhanced cleavage of poly (ADP ribose) polymerase (PARP), caspase-3 and caspase 9, Bcl-xl, and Apaf-1, as well as increased expression of death receptor 5 (DR5). We also show that the caspase-9-specific inhibitor z-LEHD-fmk inhibits drug mediated apoptosis, leading to decreased PARP and caspase-3 cleavage, and reduced DR5 expression. Importantly, through detailed studies that demonstrate the association between topoisomerase II alpha expression and DPPIV activity, our data provide further evidence of the key role played by CD26 in biological processes. PMID- 14520482 TI - Prevalence of hepatitis C virus infection among patients with hypertrophic cardiomyopathy. AB - Hypertrophic cardiomyopathy (HCM) is defined as inappropriate ventricular hypertrophy without a cardiac or systemic cause. On the other hand, hepatitis C virus (HCV) causes extrahepatic manifestations as well as chronic persistent infection in hepatocytes. We studied the association of HCV infection with HCM, comparing the prevalence of HCV antibodies between HCM patients and age- and gender-matched controls with other cardiovascular diseases at a single institution, for reasons of exclusion of bias. We then described the clinical features and genotype analysis of HCV RNA in HCM. The diagnosis of HCM was established by echocardiographic demonstration of a hypertrophied (> or =15 mm), nondilated left ventricle in the absence of another systemic or cardiovascular disease capable of producing the magnitude of hypertrophy observed. The study population consisted of 80 patients with HCM, in whom HCV antibody was examined (55 men and 25 women; mean age 56.6 +/- 12.4 years; ranging from 19 to 80 years), compared with a total of 80 age- and gender-matched controls without HCM. The prevalence of HCV infection in patients with HCM (18/80) was significantly higher than in control subjects (5/80) (Chi(2) = 7.312, P = 0.007). Of the 12 patients in whom the genotype of HCV was analyzed, 7 had type 1b and 5 had type 2a. The prevalence of HCV infection was higher in patients with HCM than in age- and gender-matched control subjects with other cardiovascular diseases. The result suggests that HCV may play an important role in these HCV-positive HCM patients. PMID- 14520481 TI - Intracranial extramedullary hematopoiesis associated with pilocytic astrocytoma: a case report. AB - Intracranial EMH is only occasionally found in primary brain tumors (mostly hemangioblastomas) and, to our knowledge, this is the first case of EMH associated with an astrocytoma. Intracranial extramedullary hematopoiesis (EMH) is described in a 29-year-old man with a recurrent pilocytic astrocytoma in the tectal region. Special stains confirmed the identities of erythroid, myeloid and megakaryocytic cells. The patient had no evidence of a predisposing bone marrow disorder or systemic EMH. Although the presence of multinucleated and blastic cells associated with a low-grade brain neoplasm is unusual, recognition of hematopoietic lineages allows EMH to be readily identified. Another tumor resection after a year of follow-up confirmed the absence of malignant progression in this recurrent astrocytoma. The small number of cases describing intracranial EMH in the absence of systemic hematologic abnormalities are correlated with the findings in this case. The low incidence of intracranial EMH indicates that cells with hematopoietic potential are seldom exposed to a supportive microenvironment within the central nervous system. However, intracranial EMH should be included as a potential, ancillary diagnosis when considering brain lesions. This may be particularly true if medical therapies involving growth factors or stem cells are found to promote hematopoiesis. PMID- 14520483 TI - The role of inflammation markers in triggering acute coronary events. AB - Studies have shown disparate results in relation to the role of plasma concentrations of inflammation markers such as fibrinogen, cytokines, and cell adhesion molecules in acute coronary syndromes. The differentiation of primary versus secondary alterations of these markers in response to acute coronary syndromes is not clear. The aim of this study was to investigate the effect of soluble cell adhesion molecules and some inflammatory markers on coronary plaque instability. The prospective study consisted of 15 patients with stable angina pectoris (SAP), 16 with unstable angina pectoris (UAP), and 16 who had undergone percutaneous transluminal coronary angioplasty (PTCA). Blood samples were obtained from the SAP group on admission, from the UAP group at the early stage of pain onset within 6 h of pain, and again after 12 h of pain. Samples from the PTCA group were collected before, 2, 14 h after the procedure. Soluble vascular cell adhesion molecule-1 (VCAM-1), endothelial selectin, interleukin-1 beta (IL-1 beta) and interleukin-2 (IL-2), and C-reactive protein (CRP) were analyzed by enzyme-linked immunosorbent assay. CRP serum levels gradually increased although IL-2 gradually decreased in patients with UAP and PTCA. In addition, VCAM-1 levels were sharply decreased after the PTCA procedure. However, this value returned back to the preprocedure levels 14 h after PTCA. Both CRP and IL-2 are directly involved in the triggering mechanisms of acute coronary events. PMID- 14520484 TI - High plasma level of asymmetric dimethylarginine in patients with acutely exacerbated congestive heart failure: role in reduction of plasma nitric oxide level. AB - Asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase, is elevated in congestive heart failure (CHF) concomitantly with the higher levels of nitric oxide (NO) and cytokines. We investigated the association among ADMA, NO, and cytokines in human CHF. Blood was collected from 25 patients with acutely exacerbated chronic CHF (acute CHF, mean age 61 +/- 3 years), 23 patients with chronic compensated CHF (chronic CHF, mean age 62 +/- 2 years), and 26 control subjects (mean age 51 +/- 1 years). ADMA was measured by high performance liquid chromatography. Tumor necrosis factor-alpha (TNF-alpha) was measured by enzyme-linked immunosorbent assay. Nitrate plus nitrite (NOx) was measured by the Griess method. The plasma levels of ADMA and TNF-Alpha were higher in patients with acute CHF than in those with chronic CHF and control subjects (both P < 0.05). The plasma level of NOx was higher in patients with chronic CHF than in those with acute CHF and control subjects (both P < 0.01). The plasma level of TNF-Alpha was positively correlated with that of ADMA in combination with patients with acute and chronic CHF (r = 0.31, P < 0.01). The plasma level of ADMA was, furthermore, negatively correlated with that of NOx (r = -0.29, P < 0.05). These findings indicate that ADMA is related to exacerbation of chronic CHF by suppression of the compensatory higher level of plasma NO. PMID- 14520485 TI - C-reactive protein-induced production of interleukin-18 in human endothelial cells: a mechanism of orchestrating cytokine cascade in acute coronary syndrome. AB - The circulating interleukin (IL)-18 level is a strong predictor of death from cardiovascular causes in patients with coronary artery disease. However, the mechanisms of IL-18 in orchestrating the cytokine cascade and the accelerator of IL-18 production in atherosclerosis are still unknown. In the present study, we measured the serum concentration of IL-18 and other markers of inflammation in 35 patients with acute coronary syndrome. To determine the mechanism of accelerating IL-18 production, we examined the release of IL-18 in human endothelial cells using human recombinant (hr) C-reactive protein (CRP) as a stimulator of IL-18. Furthermore, we investigated the inhibitory effects of hr IL-10 on IL-18 production by hr CRP in human endothelial cells. Circulating levels of IL-18 were significantly higher in patients with acute myocardial infarction than in patients with unstable angina. Incubation with hr CRP, which was equivalent to the serum concentration in patients with acute coronary syndrome, induced IL-18 release. Treatment with hr IL-10 inhibited IL-18 release in the cells stimulated with hr CRP. The serum level of IL-18 was identified as a marker of severity in acute coronary syndrome. Our findings reveal the possibility that circulating CRP by itself could cause a deterioration of the inflammatory cascade in endothelial cells associated with the upregulation of IL-18. This suggests that CRP may contribute to the mechanism of coronary artery disease in addition to being an incidental product of various types of systemic inflammation. PMID- 14520486 TI - Metoprolol, a beta-1 selective blocker, can be used safely in coronary artery disease patients with chronic obstructive pulmonary disease. AB - The coexistence of coronary artery disease (CAD) and chronic obstructive pulmonary disease (COPD) is frequent because of common etiological factors. Beta blockers remain underutilized in patients with CAD who also have COPD. This study was performed to evaluate the safety of beta-1 selective blocker agents in CAD patients with COPD. Fifty patients (aged 57.3 +/- 10.1 years) were enrolled in this study; 27 patients received metoprolol CR (controlled release), and 23 received metoprolol (conventional). The patients were stratified according to the severity of COPD (21 severe, 21 moderate, and 8 mild), started on metoprolol CR or conventional metoprolol, and titrated up to the maximum tolerated dose. The clinical controls were done during the first week and then at the first and third month. Patients received a mean total daily dose of 92.5 +/- 18 mg of metoprolol CR or 189 +/- 36.7 mg of metoprolol. Seven patients could not receive the maximum dose. There was no significant decrease in forced expiratory volume in 1 s (FEV(1)) in either group (basal vs last FEV(1): 54.5% +/- 13.4% vs 54.3% +/- 13% in the metoprolol CR group and 49.6% +/- 14.5% vs 53.2% +/- 12.8% in the metoprolol group). No adverse event was experienced. Metoprolol, a beta-1 selective blocker, can be used safely at the maximum dose in CAD patients with COPD. PMID- 14520487 TI - Changes in the peripheral eosinophil count in patients with acute eosinophilic myocarditis. AB - In many cases, the diagnosis of eosinophilic myocarditis is suggested by an elevated peripheral blood eosinophil count. However, no detailed studies have been performed on the sequential changes in the initial peripheral blood eosinophil count over the course of the disease. We measured the peripheral blood eosinophil count at the time of presentation in eight patients with eosinophilic myocarditis proven by endomyocardial biopsy and intermittently thereafter. The eosinophil count at the time of onset was <500/mm(3) in four patients, >500/mm(3) but <1,000/mm(3) in three patients, and > or =1,000/mm(3) in one patient. In three of the four patients with an initial eosinophil count of <500/mm(3), an increase to > or =500/mm(3) occurred 7-12 days after the onset. The remaining patient did not develop peripheral eosinophilia. In conclusion, in the early stage of eosinophilic myocarditis, peripheral hypereosinophilia is not present initially in some patients, and may not develop during the course of the illness in a subset of these patients. PMID- 14520488 TI - Thrombus removal with a temporary vena caval filter in patients with acute proximal deep vein thrombosis. AB - Between September 1999 and January 2001 we performed thrombus removal with the use of a temporary vena caval filter in 11 patients who had acute iliofemoral venous thrombosis. To facilitate thrombus removal, 5 patients initially received catheter-directed thrombolytic therapy (thrombolysis group), and the other 6 received surgical thrombectomy (thrombectomy group). Residual thrombus was confirmed after initial catheter-directed thrombolysis in all patients in the thrombolysis group, and thrombolysis was continued in the ward. Bleeding complications subsequently occurred in 2 patients. In the thrombectomy group, 1 patient had residual thrombus just below the temporary filter, and a permanent vena caval filter was deployed for removal. Another patient had a residual thrombus in the superficial femoral vein, and rethrombectomy was performed. One patient in the thrombectomy group died of pneumonia. All other patients were discharged. There were no deaths from pulmonary thromboembolism in this series. Post-thrombotic syndrome occurred in 2 of the 5 patients in the thrombolysis group (40%) and in 3 of the 6 patients (50%) in the thrombectomy group. We conclude that a temporary vena caval filter is useful for the management of acute proximal deep vein thrombosis, especially when aggressive treatment is required. PMID- 14520489 TI - Wave intensity analysis from the common carotid artery: a new noninvasive index of cerebral vasomotor tone. AB - Cerebral vasomotor tone is difficult to assess in patients. Wave intensity analysis has been applied to resolve complex upstream and downstream events within the vascular system. We hypothesized that the backward-traveling wave measured in the common carotid artery was caused by reflection from the cerebrovascular "beach", and that the magnitude of this reflected wave would be altered by changes in cerebral vasomotor tone. We measured common carotid arterial diameter and velocity of flow to calculate wave intensity in ten healthy male volunteers (age mean 31 +/- 3 years). Applying a rebreathing technique, we were able to increase the inspired carbon dioxide concentration to a mean of 5.9% +/- 1.7% and to compare baseline wave intensity readings to those recorded during hypercapnia. The magnitude of the reflected wave decreased significantly after CO(2) rebreathing, from -43.0 +/- 27.1 to -25.0 +/- 16.9 mmHg m s(-2), P = 0.02. This reduction in negative wave reflections in mid-systole during hypercapnia remained significant when it was analyzed as the reflection coefficient (the magnitude of the reflected wave normalized for the magnitude of the initiating forward wave, which fell from -2.8 +/- 1.5 to -1.6 +/- 1.4 ms (P = 0.01). Carotid wave reflection was significantly decreased during cerebral vasodilatation induced by increased arterial pCO(2). Wave intensity may provide a simple noninvasive means of assessing changes in cerebral vasomotor tone in vivo. PMID- 14520490 TI - Effects of cobalt-60 gamma-radiation on the synthesis of adrenomedullin and endothelin in rat vascular smooth muscle cells. AB - We studied the effects of cobalt-60 Gamma-radiation on the gene expression and secretion of adrenomedullin (Adm) and endothelin (ET) in cultured rat vascular smooth muscle cells (VSMCs). Rat VSMCs cultured in Dulbecoo's modified Eagle's medium containing 10% FBS were radiated with cobalt-60 Gamma-radiation at doses of 1, 14, and 25 Gy, respectively. Then the mRNA of Adm and ET in VSMCs were detected by the reverse-transcriptative competitive polymerase chain reaction. Adm and ET levels in rat VSMCs were measured by radioimmunoassay. As compared with that of the control, the secretions of Adm in rat VSMCs radiated at doses of 14 and 25 Gy were increased by 270% (P < 0.05) and 233% (P < 0.05), respectively. The mRNA levels of Adm were increased by 82.4% (P < 0.01) and 101% (P <0.01), respectively. Meanwhile, the secretions of ET were decreased by 27.3% (P < 0.01) and 58.0% (P < 0.01) in VSMCs radiated at doses of 14 and 25 Gy, respectively. In parallel, the mRNA levels of ET were decreased by 47.1% (P < 0.01) and 40.2% (P < 0.01), respectively. Radiation at a dose of 1 Gy had no significant effect on Adm and ET at the gene and protein levels. As compared with the control, the Adm/ET ratios in VSMCs increased by 65% (P > 0.05), 409% (P < 0.01), and 693% (P < 0.01), respectively, with radiation at doses of 1,14 and 25 Gy, respectively. The balance of Adm/ET in VSMCs could be changed by cobalt-60 Gamma-radiation, which might play an important role in the use of radiotherapy for restenosis. PMID- 14520491 TI - The morphological significance of the human sinuatrial nodal branch (artery). AB - The sinuatrial nodal branch/artery (SANB), the sinuatrial nodal branch of the coronary artery, is anatomically regarded as a significant artery since it is used as a landmark to identify the sinuatrial node, in addition to its clinical significance. In previous reports, the SANB has been shown to have 2-5 routes and it had only one branch in 91%-100% of hearts. These results indicate that compensation for the SANB is not possible in the case of its being cut or occluded. Therefore, we macroscopically reinvestigated the SANB using 106 human adult hearts to obtain a detailed understanding of its morphology. The following results and discussions were obtained from our study. (1) The SANB was observed to take six routes and two or more branches were found in 57 out of 106 cases (53.7%). (2) In those cases in which the SANB had only one branch (46.3%), it was observed to be the result of forming the proximal arterial loop in 25 cases (51.0%). The total of these cases with one branch with the proximal arterial loop between the right and left coronary arteries and those with two or more branches were 82 cases (77.4%). These results strongly suggest that compensation for the SANB could occur in the majority of cases. (3) We speculated that the SANB was generated by the disappearance of and the anastomosis between the lateral arterial loop, lateral to both auricles, and the medial arterial loop, medial to both auricles. PMID- 14520492 TI - Complications of pulmonary vein isolation by catheter ablation evaluated by ECG gated multislice computed tomography. AB - As pulmonary vein (PV) isolation by catheter ablation for paroxysmal atrial fibrillation may cause PV luminal stenosis, digital subtraction angiography or magnetic resonance imaging have been used to evaluate the lumen of the PV. Electrocardiogram-gated multislice computed tomography can evaluate the lumen of the PV from any plane desired after acquisition with excellent spatial resolution. It can also evaluate hyperplasia of soft tissue around the lumen of the PV, which cannot be evaluated by digital subtraction angiography, and may thus serve as an indicator of complications or even the effectiveness of this treatment. PMID- 14520493 TI - Regression of an atherosclerotic coronary artery plaque demonstrated by multislice spiral computed tomography in a patient with stable angina pectoris. AB - Multislice spiral computed tomography (MSCT) permits direct visualization of not only coronary artery stenosis but also atherosclerotic plaques in patients with coronary artery disease. In this report, we describe a patient with stable angina in whom the regression of the plaque was documented by serial MSCT examinations. In the patient, a 46-year-old man with stable angina, MSCT revealed a stenotic lesion at the proximal portion of the left anterior descending artery. Axial, curved multiplanar reconstruction and cross-sectional images consistently depicted a protruding computed tomography low-signal mass suggesting an atherosclerotic plaque. Intracoronary ultrasound (ICUS) also documented an eccentric soft plaque with an echo-lucent mass suggesting a lipid core. Lipid lowering therapy with pravastatin was started. Follow-up MSCT performed 7 months later documented an increase in the luminal area while the external vessel area remained unchanged. The regression of the plaque was also confirmed by a follow up ICUS study. MSCT was thought to be feasible for serial evaluation of the plaque size and texture. PMID- 14520495 TI - Spectral organization of the eye of a butterfly, Papilio. AB - This review outlines our recent studies on the spectral organization of butterfly compound eyes, with emphasis on the Japanese yellow swallowtail butterfly, Papilio xuthus, which is the most extensively studied species. Papilio has color vision when searching for nectar among flowers, and their compound eyes are furnished with six distinct classes of spectral receptors (UV, violet, blue, green, red, broadband). The compound eyes consist of many ommatidia, each containing nine photoreceptor cells. How are the six classes of spectral receptors arranged in the ommatidia? By studying their electrophysiology, histology, and molecular biology, it was found that the Papilio ommatidia can be divided into three types according to the combination of spectral receptors they contain. Different types of ommatidia are distributed randomly over the retina. Histologically, the heterogeneity appeared to be related to red or yellow pigmentation around the rhabdom. A subset of red-pigmented ommatidia contains 3 hydroxyretinol in the distal portion, fluorescing under UV epi-illumination. The red, yellow and fluorescing pigments all play crucial roles in determining the spectral sensitivities of receptors. Spectral heterogeneity and random array of ommatidia have also been found in other lepidopteran species. Similarities and differences between species are also discussed. PMID- 14520494 TI - Physiological adaptations of the raccoon dog (Nyctereutes procyonoides) to seasonal fasting-fat and nitrogen metabolism and influence of continuous melatonin treatment. AB - The raccoon dog (Nyctereutes procyonoides) is a middle-sized canid with profound autumnal fattening followed by winter sleep. This study investigated the effects of prolonged fasting-induced winter sleep on the fat and nitrogen metabolism of the species. Half of the animals were treated with continuous-release melatonin implants to induce artificial short photoperiod. Autumnal accumulation of fat was characterized by low plasma free fatty acid (FFA), diacylglycerol (DG), and triacylglycerol (TG) levels. After transition to winter catabolism, the circulating lipid levels increased due to enhanced lipolysis. Two months of fasting resulted in a steady 3.1 kg weight loss (28% of body mass, 0.47% day( 1)). Storage fat was mobilized during the winter sleep reflected by the elevated FFA and DG concentrations. The lowered insulin levels could be a stimulator for TG hydrolysis. The plasma total amino acid concentrations, urea levels, and urea creatinine ratios decreased due to fasting, whereas ammonia and total protein concentrations remained stable. The effects of melatonin on energy metabolism were modest. The results indicate that the raccoon dog is well adapted to long term wintertime fasting utilizing fat as the principal metabolic fuel. The species can maintain its protein catabolism constant for at least 60 days. Decreased cortisol and thyroid hormone concentrations may contribute to protein sparing. PMID- 14520496 TI - Long-distance navigation in the wandering desert spider Leucorchestris arenicola: can the slope of the dune surface provide a compass cue? AB - Males of the nocturnal spider Leucorchestris arenicola (Araneae: Sparassidae) wander long distances over seemingly featureless dune surfaces in the Namib Desert searching for females. The spiders live in burrows to which they return after nearly every such excursion. While the outward path of an excursion may be a meandering search, the return path is often a nearly straight line leading towards the burrow. This navigational behaviour resembles that of path integration known from other arthropods, though on a much larger scale (over tens to hundreds of meters). Theoretically, precise navigation by path integration over long distances requires an external compass in order to adjust for inevitable accumulation of navigational errors. As a first step towards identifying any nocturnal compass cues used by the male spiders, a method for detailed 3-D recordings of the spider's paths was developed. The 3-D reconstructions of the paths revealed details about the processes involved in the spiders' nocturnal way of navigation. Analyses of the reconstructed paths suggest that gravity (slope of the dune surface) is an unlikely parameter used in path integration by the L. arenicola spiders. PMID- 14520497 TI - Experience influences gustatory responsiveness to pyrrolizidine alkaloids in the polyphagous caterpillar, Estigmene acrea. AB - Electrophysiological recordings from taste sensilla of the caterpillar Estigmene acrea with the pyrrolizidine alkaloid, seneciphylline N-oxide, demonstrated that prior feeding on plants with pyrrolizidine alkaloids caused an increase in responsiveness of the PA-sensitive cells in two sensilla, relative to feeding on plants without such chemicals. Rearing on synthetic diet without pyrrolizidine alkaloids for up to seven generations caused a continuous decline in responsiveness, that could be reversed by experience with powdered Crotalaria pumila in the diet or by pure pyrrolizidine alkaloid, monocrotaline, in the diet. Response to the cardiac glycoside, ouabain, that stimulates one of the two pyrrolizidine alkaloid-sensitive cells, showed a similar decline. Pyrrolizidine alkaloids had no measurable effect on growth and development. Responses in all other taste cells were unaffected. The data are discussed in relation to the possible adaptive significance and the possible mechanisms involved. PMID- 14520498 TI - Development of highly regenerable callus lines and biolistic transformation of turf-type common bermudagrass [Cynodon dactylon (L.) Pers.]. AB - Common bermudagrass, Cynodon dactylon, is a widely used warm-season turf and forage species in the temperate and tropical regions of the world. Improvement of bermudagrass via biotechnology depends on improved tissue culture responses, especially in plant regeneration, and a successful scheme to introduce useful transgenes. When the concentration of 6-benzylaminopurine was adjusted in the culture medium, yellowish, compact calluses were observed from young inflorescence tissue culture of var. J1224. Nine long-term, highly regenerable callus lines (including a suspension-cultured line) were subsequently established, of which six were used for biolistic transformation. Five independent transgenic events, with four producing green plants, were obtained following hygromycin B selection from one callus line. Three transgenic events displayed resistance to the herbicide glufosinate, and one of these showed beta glucuronidase activity since the co-transformation vector used in the experiments contained both the gusA and bar genes. PMID- 14520499 TI - Somatic embryogenesis from leaf explants of Australian fan flower, Scaevola aemula R. Br. AB - Somatic embryogenesis from leaf explants of Scaevola aemula R. Br. was achieved. Somatic embryos were induced from explants cultured on MS medium supplemented with 0.2 mg/ 2,4-dichlorophenoxyacetic acid and 0.2-0.5 mg/l 6-benzylaminopurine (BAP). Various developmental stages of somatic embryos were found on this medium from globular embryos to germinated embryos. The transfer of globular embryos to MS medium containing 0.5 mg/l BAP resulted in a high frequency of shoot regeneration. Leaf explants cultured on MS medium containing different combinations of BAP and alpha-naphthaleneacetic acid formed adventitious shoots and roots. Histological examination confirmed the process of somatic embryogenesis. Induction of somatic embryogenesis in Scaevola provides a system for studying embryogenesis in Australian native plants and will facilitate the improvement of these plants using genetic transformation techniques. PMID- 14520500 TI - Taxonomic discrimination of higher plants by pyrolysis mass spectrometry. AB - Pyrolysis mass spectrometry (PyMS) is a rapid, simple, high-resolution analytical method based on thermal degradation of complex material in a vacuum and has been widely applied to the discrimination of closely related microbial strains. Leaf samples of six species and one variety of higher plants (Rosa multiflora, R. multiflora var. platyphylla, Sedum kamtschaticum, S. takesimense, S. sarmentosum, Hepatica insularis, and H. asiatica) were subjected to PyMS for spectral fingerprinting. Principal component analysis of PyMS data was not able to discriminate these plants in discrete clusters. However, canonical variate analysis of PyMS data separated these plants from one another. A hierarchical dendrogram based on canonical variate analysis was in agreement with the known taxonomy of the plants at the variety level. These results indicate that PyMS is able to discriminate higher plants based on taxonomic classification at the family, genus, species, and variety level. PMID- 14520501 TI - Van Buchem disease: lifetime evolution of radioclinical features. AB - OBJECTIVE: The purpose of this study was to evaluate the lifetime evolution of the radioclinical features in a large family with van Buchem disease. DESIGN AND PATIENTS: The study population included 13 patients, ranging between 6 and 69 years. The evolution of the clinical features has been assessed by retrospective analysis of the clinical records of the patients. The age-related evolution of the cortical hyperostosis and defective modeling at the tubular bones was evaluated by morphometric analysis of hand films in 9 patients, compared with 9 control individuals. Progression of sclerosis of the craniofacial bones was evaluated by analysis of the skull radiographs of eleven van Buchem patients, taken at different age. RESULTS AND CONCLUSIONS: Radioclinical features, including sclerosis of the cranial and tubular bones and cranial nerve deficit, become more prominent in older patients. Defective modeling of tubular bones, cortical thickness and medullary width progress with age. Radioclinical abnormalities of van Buchem patients become more prominent in older patients, which suggests that the van Buchem gene is very actively involved in bone metabolism throughout life. Morphometric analysis of the plain films supports the hypothesis that the physiological function of the van Buchem gene is to inhibit bone formation and possibly to regulate bone remodeling. PMID- 14520502 TI - Arcuate sign of posterolateral knee injuries: anatomic, radiographic, and MR imaging data related to patterns of injury. AB - OBJECTIVE: The "arcuate sign" is considered a pathognomonic sign for injuries of the posterolateral (PL) corner of the knee. The purpose of our study was to identify different patterns of injury to the fibular head that may associate with injuries to specific ligaments and tendons of the PL corner of the knee. The anatomic relations between the insertions of fibular collateral ligament (FCL), biceps femoris tendon (BFT), popliteofibular ligament (PFL), and arcuate ligament in normal cadaveric knees were also investigated. DESIGN AND PATIENTS: Magnetic resonance imaging was performed in two cadaveric knees which subsequently were dissected. Radiopaque markers were placed upon the fibular insertions of the FCL, BFT, PFL, and arcuate ligament in the dissected knees, and knee radiographs were then obtained. Twelve patients with radiographic or MR imaging evidence of isolated injury to the PL corner of the knee were retrospectively reviewed, with regard to avulsion fractures and marrow edema in the fibular head and the integrity of the ligaments of the PL corner of the knee. RESULTS AND CONCLUSIONS: The PFL and arcuate ligament were seen to attach directly to the posterior and medial aspect of the styloid process of the fibular head. The FCL and BFT attached as a conjoined structure on the lateral aspect of the fibular head lateral, anterior and inferior to the attachment site of the PFL and arcuate ligament. Injury to the arcuate ligament or PFL was diagnosed in 8 patients who presented with a small avulsion fracture of the styloid process of the fibula (n=2), bone marrow edema in the medial aspect of the fibular head (n=3), or both (n=3). In 4 patients with injury to the conjoined tendon or FCL, a larger avulsion fragment and more diffuse proximal fibular edema were seen. Radiographic and MR imaging findings in injuries of the posterolateral corner of the knee may suggest injury to specific structures inserting in the fibular head. PMID- 14520503 TI - Initial characterization of a protochordate histocompatibility locus. AB - The colonial protochordate, Botryllus schlosseri, undergoes a natural transplantation reaction which is controlled by a single, highly polymorphic locus called the Fu/HC. We are using map-based cloning to identify Fu/HC gene(s), and have currently delineated their location to an approximately 1-cM region of the B. schlosseri genome. The Fu/HC physical map currently consists of 85 sequence-tagged sites mapped on a minimum tiling path of 800 kb which consists of five contigs, with four gaps remaining to be crossed, and is estimated to be 75% completed. Approximately half this region has been sequenced throughout the locus, allowing the first analysis of a metazoan histocompatibility locus outside of vertebrates. This has resulted in the identification of 18 predicted genes, a number of which have been found to be expressed. Several of these genes are well conserved among the chordates; however, none of the predicted or expressed genes are linked within the genome of any organism in the databases. In addition, the Fu/HC is one of the most polymorphic loci ever described, and physical mapping has revealed that the locus is quite dynamic, and includes features such as hotspots of recombination. PMID- 14520505 TI - Reversible acute renal failure after unilateral extracorporeal shock-wave lithotripsy. AB - Extracorporeal shock-wave lithotripsy (ESWL) is the treatment of choice for the majority of patients with renal or proximal ureteral stones. We describe an unusual case of anuric renal failure following ESWL, in absence of obstruction or myoglobinuria, in the presence of unilateral nephrolithiasis and two normally functioning kidneys. A mechanism for this patient's acute renal failure (ARF) is postulated. Although the frequency of ARF after ESWL is extremely rare and the mechanism responsible for ARF is not understood, the appearance of ARF, when ureteral obstruction or hematoma are absent, should be included among complications following EWSL. Attention should be paid to older patients. PMID- 14520504 TI - An experimental model for stricture studies in the anterior urethra of the male rabbit. AB - In this study, an animal model was developed for the examination of urethral strictures (US). Through a resectoscope, a resection was made in the urethras of 15 male rabbits. After 30 days, the rabbits were evaluated with urethrography, impedance planimetry and either histology or the determination of collagen content. Fifteen rabbits serving as controls were evaluated in the same way. Three rabbits in the resection group and one in the control group died before evaluation. Urethrography demonstrated a stricture in the remaining 12 animals in the resection group. The urethras of the control animals were all normal. Impedance planimetry confirmed that the luminal cross sectional area (CSA) of the strictures was significantly smaller than the CSA of the corresponding part of the urethra in the control group. No difference in CSA was found 1 cm proximal to the stricture. The strictures consisted of densely woven collagen which sent tongues into the adjacent normal parts of the urethra. No difference in collagen content was found between the two groups either at the stricture site or 1 cm proximally. The described method of producing US in the rabbit model was very consistent with all operated animals developing a stricture. The model might prove valuable in evaluating new methods for the treatment of US. PMID- 14520506 TI - Existence of functional M3-muscarinic receptors in the human heart. AB - It has been recently shown that, in adult rat ventricular cardiomyocytes, functional muscarinic receptors (M-receptors) of the M(3)-subtype exist that mediate inositol phosphate (IP) formation. The aim of this study was to characterize the M-receptor subtype mediating IP formation in the human heart. For this purpose in [3H]-myo-inositol labeled slices of human right atria, carbachol-induced [3H]-IP formation and its inhibition by several M-receptor antagonists was assessed. Carbachol (0.1 microM-100 microM) increased [3H]-IP formation; maximal increase at 100 microM was 93+/-16% above basal ( n=20); the pEC(50)-value for carbachol was 5.56. Atropine (1 microM) completely suppressed 100 microM carbachol-induced [3H]-IP formation. Among the M-receptor subtype "selective" antagonists himbacine (1 microM) and pirenzepine (1 microM) only marginally affected carbachol-induced [3H]-IP formation whereas the M(3)-receptor antagonist darifenacin (1 nM-1 microM) concentration-dependently inhibited carbachol-induced [3H]-IP formation with a pK(i)-value of 8.49. We conclude that in human right atrium there exist functional M(3)-receptors that couple to IP formation. PMID- 14520507 TI - Genotoxicity of benomyl and its residues in somatic and germ cells of mice fed on treated stored wheat grains. AB - Swiss mice were fed for 2, 4 and 8 weeks wheat grains treated with 1, 2, and 4 g benomyl/kg and stored for 6 and 12 weeks. The maximum effect of benomyl on the induction of chromosomal aberrations was observed after feeding mice for 8 weeks with wheat grains treated with 4 g benomyl/kg and stored for 12 weeks. Its proportion differed significantly in bone marrow and spermatocyte cells, 15+/ 0.51% vs. 13.4+/-0.66%, respectively, from that in nontreated mice (background level), 4.4+/-0.24% and 3.8+/-0.20%, respectively. Lengthening the storage period of treated wheat grains caused a dose-dependent increase in the frequency of sister chromatid exchanges: 8.61+/-0.34 vs. 4.16+/-0.06/cell. The proportion of sperm-head abnormalities increased by lengthening the period of storage and feeding: 7.7+/-0.41% vs. 3.25+/-0.12%. In another experiment mice were orally treated by gavage with benomyl at 50, 100, 150, 200 mg/kg; a significant and dose dependent increase in sperm-head abnormalities was observed. These findings demonstrate that benomyl (a 50% wettable powder formulation) and its residues in wheat grains are genotoxic in mice. PMID- 14520508 TI - Hepatotoxin-induced hypercreatinaemia and hypercreatinuria: their relationship to one another, to liver damage and to weakened nutritional status. AB - Hypercreatinuria is a well-known feature of liver and testicular toxicity and we have recently proposed that hepatotoxin-induced hypercreatinuria would arise as a consequence of increased cysteine synthesis associated with the provision of protective substances (glutathione and/or taurine). Here a direct relationship between hepatotoxin-induced hypercreatinaemia and hypercreatinuria is shown and the possible relationships of hepatotoxin-induced hypercreatinaemia and hypercreatinuria to hepatic damage and to weakened nutritional status are examined. Male Sprague-Dawley rats were dosed with a variety of model hepatotoxins at two dose levels per toxin. Blood plasma samples taken at 24 h post-dosing and urine samples collected from 24-31 h post-dosing were analysed by (1)H NMR spectroscopy. Both hypercreatinaemia and hypercreatinuria were found in rats dosed with allyl formate (75 mg/kg), chlorpromazine (30 and 60 mg/kg), alpha naphthylisothiocyanate (ANIT, 100 mg/kg) and thioacetamide (200 mg/kg), whilst significant hypercreatinuria, but not hypercreatinaemia, was found after dosing with thioacetamide (50 mg/kg). Neither hypercreatinaemia nor hypercreatinuria were found after dosing with allyl formate (25 mg/kg), ethionine (300 and 1000 mg/kg) or ANIT (30 mg/kg). Reduced feeding is known to cause hypercreatinuria in rats and, of the four hepatotoxins that induced hypercreatinaemia and hypercreatinuria at the given time-points, two, chlorpromazine and ANIT, also affected nutritional status with ketosis being clearly identifiable from the plasma (1)H NMR spectra. Thus, the creatine changes induced by ANIT and chlorpromazine are potentially attributable, in whole or in part, to reduced feeding rather than to liver effects alone and, consequently, the results were examined with and without inclusion of the ANIT and chlorpromazine data. With all of the data included, there were eight out of ten points of correspondence between the incidence of hypercreatinaemia and/or hypercreatinuria and the incidence of increases in plasma alanine aminotransferase (ALT) activity. At the same time there were nine out of ten points of correspondence between the incidence of hypercreatinaemia and/or hypercreatinuria and the incidence of increases in plasma aspartate aminotransferase (AST) activity. However, with the ANIT and chlorpromazine data excluded there was complete (six out of six points) correspondence between the incidence of hypercreatinaemia and/or hypercreatinuria and the incidence of increases in plasma AST and ALT in the remaining data. Likewise, with all of the data included, there was some apparent correlation (correlation coefficient, r=0.80) between the group mean levels of plasma AST and plasma creatine when expressed relative to the mean values for controls sampled at the same time-point. However, with the ANIT and chlorpromazine data excluded, that correlation coefficient was increased to 0.95. The findings of these studies suggest that the ANIT- and chlorpromazine-induced creatine changes may have been caused by reduced feeding rather than by liver toxicity. The allyl formate and thioacetamide data indicate that hepatocellular necrosis is accompanied by increases in plasma and urinary creatine, and suggest the possibility of a quantitative relationship between the increases in plasma AST and the increases in plasma creatine that are associated with hepatocellular necrosis. The ethionine and ANIT data suggest that fatty liver (steatosis) and cholestatic damage may not be associated with hypercreatinaemia and hypercreatinuria. PMID- 14520509 TI - Lack of maternal dietary exposure effects of bisphenol A and nonylphenol during the critical period for brain sexual differentiation on the reproductive/endocrine systems in later life. AB - Two potential endocrine-disrupting chemicals, bisphenol A (BPA) and nonylphenol (NP), were assessed for their long-lasting effects on endocrine/reproductive systems following transplacental and lactational exposure to rat offspring during a time-window that included the critical period for brain sexual differentiation. Each chemical was mixed with diet at concentrations of 60, 600 and 3000 ppm and was provided to maternal Sprague-Dawley rats from gestational day (GD) 15 to postnatal day (PND) 10. Ethinylestradiol (EE) at 0.5 ppm was used as an estrogenic reference drug. During pregnancy and lactation, including the exposure period, a soy-free rodent diet was provided to eliminate possible modification of the study results by plant-derived phytoestrogens. Effects on endocrine/reproductive systems were evaluated by examining the anogenital distance, organ weights before puberty, onset of puberty, estrous cyclicity, and organ weights and histopathology of adult endocrine organs (at 11 weeks of age), as well as the volume of the sexually dimorphic nucleus of preoptic area. Both NP and BPA, at high doses, caused decreases in maternal body weights and retardation of offspring growth, but neither affected any of the endocrine/reproductive endpoints of offspring, whereas EE induced irreversible changes in estrous cyclicity and histopathology of ovaries and uterus of adult females. The results indicated that maternal dietary exposure to NP or BPA at concentrations up to 3000 ppm from GD 15 through PND 10 do not exert any apparent adverse effects on the endocrine/reproductive systems of offspring. PMID- 14520510 TI - Electrophilic tissue burden in male Sprague-Dawley rats following repeated exposure to binary mixtures of polycyclic aromatic hydrocarbons. AB - The aim of this study was to investigate the electrophilic tissue burden (ETB) formation, assessed as covalent binding of the ultimate carcinogen benzo( a)pyrene diolepoxide (BaPDE) with cellular proteins, in liver, lung and heart, as well as with haemoglobin (Hb) following repeated exposure to binary mixtures of benzo( a)pyrene (BaP) and pyrene (P). Male Sprague-Dawley rats were injected intraperitoneally, once daily for 10 consecutive days, with binary mixtures of BaP and P in three different exposure scenarios corresponding to BaP/P ratios of 0.2, 1 and 5 and with three dose levels of BaP (2, 6 and 20 mg/kg) for each scenario. ETB levels were measured as the ultimate analyte benzo( a)pyrene tetrol (BaPTeT) obtained after mild acid hydrolysis of BaPDE-adducts with proteins. A high-performance liquid chromatography fluorescence technique was used to quantify the analyte. Similar ETB levels (within a factor of 4) were observed in all tissues studied at any given binary dose. However, the ETB generally tended to be somewhat higher in metabolically active tissues (i.e. liver and lung) than in metabolically non-active tissues (i.e. heart and Hb). Lack of influence of pyrene on ETB levels in all tissues was confirmed over the binary dose range examined. Linear BaP-dose-dependent ETB formation in all tissues (at P20 per 1000 woman years) from a group at low risk (i.e. below the average risk in the cohort). QUS screening alone has a very low sensitivity (15%). The strategy using QUS as a method of triage and that combining selective BMD and clinical assessment have a sensitivity equivalent to systematic BMD screening (around 35%), with less than 50% BMD examinations. The high-risk women identified by these two strategies are not the same. A simple algorithm combining QUS, BMD, and clinical risk assessment allows an increased number of high-risk women to be identified (21%), and thus improves the sensitivity (53%). With this combined strategy, women in the high risk group have one chance in ten of having a hip fracture over the next 4 years, whereas women in the low risk group have only one chance in 40. PMID- 14520512 TI - Is vasculo-neural ingrowth the cause of pain in chronic Achilles tendinosis? An investigation using ultrasonography and colour Doppler, immunohistochemistry, and diagnostic injections. AB - The purpose of this study was to investigate where the pain comes from in chronic mid-portion Achilles tendinosis, by using ultrasonography and colour Doppler, immunohistochemistry, and diagnostic injections. Twenty-five tendons in 24 patients (mean age 47 years) with the clinical diagnosis of painful chronic mid portion Achilles tendinosis, and 20 tendons in 14 controls (mean age 48 years) with pain-free Achilles tendons were examined with ultrasonography and colour Doppler. For the 25 painful tendons, a local anaesthetic was injected in the area with neovascularisation outside the ventral part of the tendon. Biopsies taken from tendinosis tissue in six patients were used for PGP 9.5 immunohistochemistry. A neovascularisation, inside and outside the ventral part of the area with tendon changes, was demonstrated in all painful tendinosis tendons, but not in any of the pain-free control tendons. After injection of the local anaesthetic towards the neovessels outside the ventral part of the tendon, the pain during tendon-loading activity was temporarily cured in all patients. The mean VAS-score for heel-raises decreased significantly from 75 mm to 6 mm. In biopsies, PGP 9.5 immunohistochemistry showed nerve structures in the vicinity of blood vessels. In conclusion, we demonstrate findings which support neovessels and accompanying nerves being the possible source of pain in chronic mid-portion Achilles tendinosis. PMID- 14520513 TI - Efforts to standardize animal sensitization in asthma research. Scientific cooperation - quo vadis? PMID- 14520514 TI - How do statins control neuroinflammation? AB - Statins are used widely to reduce the levels of low-density lipoproteins and cholesterol and thereby lower the incidence of atherosclerosis and cardiovascular disease. They achieve this through their ability to limit the production of mevalonate via blockade of 3-hydroxy-3-methyl glutaryl coenzyme A (HMG-CoA) reductase. This enzyme is the rate limiting step controlling the de novo production of cholesterol. It is now clear from a number of studies in various animal models of neuroinflammation as well as in in vitro cell trafficking studies, that statins significantly attenuate both the clinical symptoms of neuroinflammation and the associated infiltration of inflammatory cells into the CNS. It has been known for some time that statins have additional effects, which appear independent of their cholesterol lowering action. Although the precise mechanism by which statins are able to exert this inhibitory effect on leukocyte infiltration and consequential neuroinflammatory disease is presently unclear, a number of potential mechanisms have been proposed. PMID- 14520515 TI - The production of anti-inflammatory cytokines in whole blood by physico-chemical induction. AB - OBJECTIVE AND DESIGN: Cytokines such as interleukin-1 (IL-1) play an important role in degenerative musculo-skeletal diseases, including osteoarthritis, and a multitude of inflammatory disorders. Agents that inhibit the action of such cytokines have a high therapeutic potential in such diseases. Here we describe a new method for enhancing the production of the interleukin-l receptor antagonist (IL-1Ra) and other anti-inflammatory cytokines in whole blood. MATERIAL AND METHODS: Human venous blood was incubated in the presence of CrSO(4)-treated glass beads. Serum was recovered and the concentrations of IL-1Ra and other relevant cytokines were measured by ELISA. RESULTS: The interaction of the glass bead surface with cells in whole blood increased production of IL-1Ra and anti inflammatory cytokines. Removal of the beads and centrifugation generated a serum preparation enriched in anti-inflammatory cytokines. This preparation is of therapeutic value in treating various inflammatory and degenerative disorders. CONCLUSIONS: The increased de novo production of anti-inflammatory cytokines by a direct physico-chemical induction of whole blood in the Orthokin system is feasible and offers an alternative, novel approach to treating mild to moderate OA and other orthopaedic conditions such as degenerative spine diseases. PMID- 14520516 TI - Early and late histamine release induced by albumin, hetastarch and polygeline: some unexpected findings. AB - OBJECTIVE: The perioperative use of colloidal plasma substitutes is still under discussion. We therefore conducted a prospective randomised study with three commonly used plasma substitutes to examine their histamine releasing effects in 21 volunteers. MATERIAL OR SUBJETS: 21 male volunteers were enrolled in this prospective, randomised, controlled clinical study. Endpoints were the incidence of early and late histamine release and the time course of the release kinetics. Normovolemic hemodilution technique was used with hydroxyethyl starch (n = 6), human albumin (n = 6) and polygeline (n = 9). Measurement and observation period was 240 min after the start of the plasma substitute infusion. Heart rate, blood pressure, SaO(2), clinical symptoms/signs and plasma histamine were measured during the observation period. RESULTS: The incidence of histamine release over the whole observation period in all three groups was 100%. Histamine release occurred frequently in all three groups until 30 min (50%-78%) and up to 240 min (late release reaction: 67%-83%) after the start of infusion. Surprisingly even hydroxyethyl starch, which is regarded as a generally safe and effective plasma substitute, caused high incidences of late histamine release (67%). Histamine release is a well known side effect of polygeline and - to a lesser extent - also of albumin, but was a novel finding for hydroxyethyl starch. CONCLUSIONS: We demonstrated for the first time histamine releasing effects of hydroxyethyl starch over a long period of time after administration. This perioperatively and for intensive care possibly relevant finding should make clinicians aware of late side effects not yet connected with the clinical use of these colloidal plasma substitutes. PMID- 14520517 TI - Analysis of histamine and modeling of ligand-receptor interactions in the histamine H(1) receptor for Magic Angle Spinning NMR studies. AB - OBJECTIVE AND DESIGN: Investigation of the principles of ligand-receptor interaction in histamine receptors can help to provide a solid foundation for structure-based drug design. Stable isotope labelling of the ligand 'Histamine' has been performed and 1D (13)C CP MAS and 2D Radio Frequency Dipolar Recoupling (RFDR) spectra for the ligand are presented. Hyperfine signals were well spread and did not suffer from any sizable line broadening. The production of H(1) receptor for Magic Angle Spinning NMR studies is currently in progress. TREATMENT: An agonist binding domain is proposed using homology modeling, database searches and mutagenesis data for the H(1) receptor. METHODS: Homology modeling, Database searches for Expressed sequence Tag (ESTs), Magic Angle Spinning Nuclear Magnetic Resonance analysis of the ligand histamine. RESULTS: The three-dimensional receptor model and mutagenesis studies suggest that the amine of the agonist histamine may form an ion pair with the TM III Asp, whereas the imidazole ring of histamine may associate with TM V Asp and Thr. CONCLUSIONS: Homology modeling studies confirms the absence of TM VIII in the H(1) receptor. According to the model the histamine in particular interacts with the transmembrane (TM) regions of the H(1) receptor structure, in particular TM helix III and V. This is in line with recent mutagenesis studies. Database search methods for ESTs have been used for electronic prediction of tissue distribution of H(1) receptor expression. The results indicate that the H(1) expression is highest in heart and skeletal muscle, which may be of importance for drug targeting. PMID- 14520518 TI - Comparison of the effects of desloratadine and levocetirizine on histamine induced wheal, flare and itch in human skin. AB - OBJECTIVE: A previous study showed the inhibitory effects of loratadine on histamine-induced wheal, flare and itch in human skin to be very variable between individuals. It was hypothesised that this variability may have been due to differences in the rates of metabolism of loratadine to its active form, desloratadine. This double blind, crossover study examined the effects of desloratadine in 12 healthy volunteers. Levocetirizine was used as a comparator. METHODS: Desloratadine (5 mg), levocetirizine (5 mg) or placebo was taken orally 4 h before an intradermal injection of histamine (20 microL, 100 microM) or vehicle control into the forearm skin. Flare areas were assessed by scanning laser Doppler imaging before and at 30 s intervals for a period of 9 min. Wheal areas were measured by planimetry at 10 min. Itch was scored every 30 s for 5 min using a visual analogue scale. RESULTS: Following placebo administration, the mean (+/- SEM) wheal area at 10 min was 79.3 +/- 6.9 mm(2), mean flare area for the first 5 min following challenge 26.6 +/- 2.7 cm(2), and itch score for the same period 48.5 +/- 7.6%. The effects of desloratadine were variable between individuals, mean reductions in the wheal and flare areas being 17% (P = 0.033) and 12% (P = 0.036). Desloratadine did not reduce itch significantly. Levocetirizine was more consistent in its effects, mean reductions in wheal, flare and itch being 51%, 67% 78% respectively (all P < 0.001). CONCLUSIONS: A single dose of 5 mg levocetirizine produced more consistent and greater inhibitory effects on histamine-induced wheal, flare and itch than did 5 mg desloratadine. The difference is suggested to reflect the basic pharmacokinetics of the two drugs. PMID- 14520519 TI - Monocyte chemoattractant protein (MCP)-1 production via functionally reconstituted Fcalpha receptor (CD89) on glomerular mesangial cells. AB - BACKGROUND: Fc alpha receptor (FcalphaR; CD89) is the receptor for Fc portion of IgA in various cells, and displays various immunological responses on binding. It is important to analyze the mesangial functions via FcalphaR in the pathogenesis of IgA nephropathy. However, it is still controversial whether FcalphaR is expressed on mesangial cells. To assess biological functions of FcalphaR on the mesangial cells, we established mesangial transfectants that expressed FcalphaR with or without FcRgamma chain that is a common signaling molecule of FcRs. The production of monocyte chemoattractant protein-1 (MCP-1) by mesangial cells is known to contribute to cellular infiltration into glomeruli and subsequent glomerular injuries. METHODS: Murine mesangial cell lines (SV40 MES 13) were transfected with cDNA of the human FcalphaR. Furthermore, we co-transfected some of the FcalphaR transfectants with cDNA of human FcRgamma chain. The tyrosine phosphorylation of the intra-mesangial proteins after FcalphaR cross-linking was examined by immunoprecipitation. MCP-1 production from each transfectant stimulated with heat aggregated IgA was determined by sandwich ELISA. RESULTS: Two kinds of mesangial transfectants stably expressed human FcalphaR with or without FcRgamma chain (FcalphaR(+), FcalphaR(+)/gamma(+)). Phosphorylation of FcRgamma chain and syk kinase was detected in FcalphaR(+) and FcalphaR(+)/gamma(+) cells, but not in untransfected cells. Aggregated IgA induced significantly higher MCP-1 production in FcalphaR(+)/gamma(+) than those in FcalphaR(+) or untransfected control. CONCLUSIONS: Present study demonstrated that FcalphaR and FcRgamma chain could be reconstituted in mesangial cells and mediated MCP-1 production by aggregated IgA in a dose-dependent manner. Current data would argue that FcalphaR can be activated in mesangial cells through their own machinery, although underlying mechanisms for FcalphaR induction in mesangial cells remain unclear. PMID- 14520520 TI - Perioperative elastase activity in cardiac surgery and its role in endothelial leakage. AB - OBJECTIVE AND DESIGN: The functional activity and pathophysiological effects of polymorphonuclear elastase (PMNE) in cardiac surgery patients are unknown. This in vitro study was done to evaluate whether PMNE activity in patient blood samples may be correlated with decreased endothelial wall integrity. METHODS AND SUBJECTS: PMNE was serially analyzed by PMNE activity in plasma samples from 40 high risk cardiac surgery patients. Endothelial cell cultures were used to study the influence of patient serum on the intercellular integrity. RESULTS: Ex vivo, samples with high PMNE activity (>1.0 mg/ ml), found in 14 patients (35%), neither induced hyperpermeability in cultured endothelial cells nor resulted in intracellular redistribution of the junction molecules cadherin-5 or beta catenin. However, pretreatment of endothelial cells with these samples but not with low activity (<0.5 mg/ml) samples augmented neutrophil transendothelial migration (>20-fold) in conjunction with formation of intercellular gaps and irregular membrane-associated beta-catenin staining. Neutrophil transmigration was inhibited by blocking neutrophil beta1 integrin but not by the proteinase inhibitor methoxysuccinyl-Ala-Ala-Pro-Ala. CONCLUSIONS: Augmented PMNE activity in cardiac surgery patients does not directly induce endothelial leakage but may indirectly promote neutrophil extravasation and thus perioperative endothelial hyperpermeability. PMID- 14520521 TI - Detection of myocardial viability in acute infarction using contrast-enhanced (1)H magnetic resonance imaging. AB - BACKGROUND: Reperfusion strategies salvage myocardium at risk in acute myocardial infarction (MI). This clinical study was performed to determine whether areas without evidence of delayed MRI contrast enhancement in MI correspond to viability by means of percent systolic wall thickening (%SWT) and enddiastolic wall thickness (EDWT) in chronic infarction. METHODS: Twenty MRI studies were performed in ten patients within 6 days of MI and 3 months post-MI. On a segmental basis the percentage of viable myocardium as defined by contrast enhanced MRI (no delayed MRI contrast enhancement) in acute MI was measured and was compared with %SWT and EDWT in chronic MI. RESULTS: Of the 1718 segments in acute infarction in which the percentage of viable myocardium was measured 1333 were found to be completely viable by means of contrast-enhanced MRI (no delayed MRI contrast enhancement). All of these segments revealed %SWT on day 90 post-MI, and 97% of segments were viable by means of an EDWT of more than 5.5 mm. In 85 segments the proportion of viable myocardium was 50-99% (mean 56+/-8%), with 92% segments found to be viable by means of %SWT and 92% by EDWT, and of 156 segments with viable myocardium between 1-49% (36+/-8%) 79% were found to be viable by means of %SWT and 82% by EDWT. Corresponding proportions of 144 segments with transmural delayed MRI contrast enhancement in acute MI were 45% and 17%. CONCLUSIONS: In acute reperfused MI viable myocardium as delineated by contrast enhanced MRI is correlated with clinical parameters of viability. Delayed MRI contrast enhancement resolves nontransmural MI and may become a valuable clinical tool when planning revascularization procedures. PMID- 14520522 TI - Dorsal foramenal extraosseous epidural cavernous hemangioma. AB - Cavernous hemangiomas are vascular hamartomatous malformations that affect the central nervous system. This pathology is frequently encountered in the cerebral hemispheres, cerebellum, and brainstem. Cavernous hemangiomas infrequently occur at the spinal level or relative to an intramedullary localization; extramedullary epidural sites are also sometimes affected by this pathology. We report an extradural, extramedullary, cavernous hemangioma with foramenal extension of the dorsal section and discuss the differential diagnosis with dorsal foramenal neurinoma. A 52-year-old woman was admitted with irritation and deficit symptoms radiating into the left D3 spinal root space. The first radiological diagnosis was of a foramenal neurinoma beginning at the D3 root. The presence of a heterogeneous MR signal in both T1 and T2 images led us to consider the differential diagnosis of a cavernous hemangioma lesion. The patient underwent microsurgical treatment with a far lateral extraforamenal approach. Symptoms quickly improved: pain and dysesthesia disappeared after surgery and only light hypoesthesia was found. We want to stress the importance of MR imaging in formulating a correct differential diagnosis with foramenal neurinoma and underline that microsurgical treatment with a far lateral extraforamenal approach allowed us to remove the lesion completely without affecting Denis's posterior column, the lamina, and the articular facet. PMID- 14520523 TI - Construction and characterization of a soybean yeast artificial chromosome library and identification of clones for the Rps6 region. AB - We report the construction and characterization of the first soybean yeast artificial chromosome (YAC) library using high-molecular weight DNA isolated from leaf nuclei of the cultivar Conrad 94 that carries Phytophthora resistance genes Rps1-k and Rps6. The quality of this library has been evaluated through analysis of 393 randomly selected YAC clones. The library consists of 36864 clones, of which approximately 19956 carry single soybean YACs with an average size of about 285 kb. The library represents approximately five soybean genome equivalents. The probability of finding any soybean sequences from this library is about 0.99. The library was screened for 43 SSR markers representing the whole soybean genome. We were able to identify positive YAC pools for 95% of the SSR markers. Two YAC clones carrying molecular markers linked to the Rps6 gene were identified. The YAC library reported here would be a useful resource for map-based cloning of agronomically important soybean genes and also to complement the effort towards construction of the physical map for the soybean genome. PMID- 14520524 TI - Neoadjuvant chemotherapy: a standard treatment for locally advanced gastric cancer in the near future? PMID- 14520525 TI - Quantitative evaluation of reconstruction methods after gastrectomy using a new type of examination: digestion and absorption test with stable isotope 13C labeled lipid compound. AB - BACKGROUND: Digestive and absorptive disorders may negatively influence patients' nutrition, thus resulting in weight loss after gastrectomy. A relationship thus seems to exist between the fat absorptive function and body weight after gastrectomy; however, so far there has been no evidence to prove this hypothesis. Therefore, in this study we evaluated fat absorption ability using a stable isotope, (13)C-trioctanoin, based on the range of the gastrectomy and the method of reconstruction, and we also determined the feasibility of this test. METHODS: Among patients who had undergone gastrectomy for gastric cancer, 40 patients who had been operated on between 1 and 3 years previously were evaluated. Ten patients had undergone the double-tract (DT) method, and 10 patients had received the Roux-en-Y (RY) method after a total gastrectomy. Twenty patients who had undergone the Billroth I (BI) method after a distal gastrectomy were the control group. In addition, 10 volunteers formed a healthy control group for the (13)C trioctanoin test. We also examined other six factors related to nutrition after gastrectomy. RESULTS. The (13)C-trioctanoin test showed, in relation to the reconstruction procedure, the highest average peak of fat absorption in the BI group (which had food passage through the duodenum), followed by the average peak of fat absorption in the DT group and the RY groups. In a comparison of duration, at 60 min and 90 min after administration, the BI group and DT group showed a significantly higher level than the RY group. The peaking time (average time at peak level) showed a significant difference between the RY group and the other groups. The absorption amount at an early stage of absorption and the percent (%) dose showed a significant difference between the RY group and the other groups. The RY group had significantly lower fat absorption than the healthy controls. CONCLUSION: According to this study, which evaluated fat absorption after different reconstructive procedures after gastrectomy, the procedure that accommodated for the passage of food through the duodenum showed better results for the absorption of medium-chain triglycerides, and the patients also showed a better physiological state. PMID- 14520526 TI - The usefulness of CEA and/or CA19-9 in monitoring for recurrence in gastric cancer patients: a prospective clinical study. AB - BACKGROUND: Many studies on postoperative carcinoembryonic antigen (CEA) and/or carbohydrate antigen (CA)19-9 monitoring after operation for gastric cancer have been reported, but most have been retrospective. METHODS: A nationwide observational study was implemented in 135 leading institutions in Japan to evaluate the significance of CEA and/or CA19-9 in postoperative monitoring for recurrence in patients with advanced gastric cancer. Three hundred and twenty-one patients examined in this analysis underwent radical gastrectomy at one of Japan's leading institutions between November 1993 and March 1996 and had been followed up for at least 5 years. Serum levels of CEA and CA19-9 were examined preoperatively and every 3 months postoperatively, with diagnostic imagings, such as chest X-ray, computed tomography (CT), and ultrasonography also being performed every 3 months. RESULTS: Recurrence was observed in 120 patients (peritoneum, 48; liver 16; lymph node, 16; multiple sites, 25; and others, 12). Sensitivities of CEA and either CEA or CA19-9, or both, for recurrence were 65.8% and 85.0%, respectively, both of which values were significantly higher than the preoperative positivities (28.3% and 45.0%, respectively). In most patients with high preoperative levels CEA and/or CA19-9, these tumor markers increased again at recurrence. Recurrent diseases were detected between 5 months after detection by diagnostic imagings and 12 months before detection by diagnostic imagings (mean of 3.1 +/- 3.6 months before detection by diagnostic imagings) and between 10 months after detection by diagnostic imagings and 13 months before detection by diagnostic imagings (mean of 2.2 +/- 3.9 months before detection by diagnostic imagings) by CEA and CA19-9 monitorings, respectively. CONCLUSION: These results suggest that CEA and/or CA19-9 monitoring after operation was useful to predict the recurrence of gastric cancer, especially in almost all the patients with high preoperative levels of these markers. PMID- 14520527 TI - A new scoring system for peritoneal metastasis in gastric cancer. AB - BACKGROUND: The degree of peritoneal metastasis (PM) of gastric cancer has been classified into four grades (P0, P1, P2, and P3) in the Japanese Classification of Gastric Carcinoma, First English Edition. However, the PM category in this version was so ambiguous in terms of volumetry that it was unlikely to estimate PM status correctly. We have developed a new volume scoring system for PM, modifying Sugarbaker's classification. METHODS: Eighty-five patients with gastric cancer with PM underwent surgery and/or laparoscopy at our hospital at Kanazawa University between January 1990 and December 2000. The grading (Gr) by the volume scoring system was decided according to the sum of scores for nine (in male) or ten (in female) abdominopelvic regions. These regions included left subphrenics, right subphrenics, left flank, right flank, pelvis, omenta, mesentery, small and large bowels, ovaries (only for females), and other intraabdominal organs. Volumetry for each region was carried out using the following categories: VS0, indicating the absence of cancer in a particular abdominopelvic region; VS1, indicating that tumor nodules are less than 10 mm in diameter; VS2, indicating tumors between 1 and 5 cm in diameter; and VS3, indicating that tumor is more than 5 cm in diameter or that an organ is coated by a mat of tumor (confluent disease). The scores for VS0, VS1, VS2, and VS3 were 0, 1, 2, and 3 points, respectively. Total scores of 1 to 3 were the equivalent of Gr I, total scores of 4 to 6 were equivalent to Gr II, total scores of 7 to 9 were equivalent to Gr III, and total scores of more than 9 were equivalent to Gr IV. RESULTS: There was no significant difference between P1 and P2, or between P1 and P3 in survival curves. But the 50% survival times of Gr I, Gr II, Gr III, and Gr IV were 1.23, 0.66, 0.67, and 0.32 years, respectively. The prognosis of Gr I was significantly better than that of any other grades. Eight of the 28 patients (29%) assessed as Gr I survived more than 2 years, whereas only 2 of the 57 (4%) assessed as Gr II, Gr III, or Gr IV did so. Multivariate analysis by Cox's proportional hazard model revealed that the volume scoring, resection of primary tumor, and resection of peritoneal tumor were independent factors to predict prognosis. CONCLUSION: These results showed that the volume scoring system was strongly related to the prognosis, and the patients assessed as Gr I had a great probability of cure by surgery. PMID- 14520528 TI - Biweekly administration regimen of docetaxel combined with CPT-11 in patients with inoperable or recurrent gastric cancer. AB - BACKGROUND: Both docetaxel (TXT) and irinotecan (CPT-11) are active chemotherapeutic agents for gastric cancer. We designed a biweekly administration regimen of TXT combined with CPT-11 for 4 weeks as one cycle in patients with inoperable or recurrent gastric cancer, and conducted a dose-escalation study. METHODS: Patients with histologically confirmed gastric cancer were treated with the regimen. The dosage levels of TXT and CPT-11 were as follows: level 1, 30 mg/m(2) and 50 mg/m(2); level 2, 35 and 50 mg/m(2); level 3, 40 and 50 mg/m(2); level 4, 40 and 60 mg/m(2); and level 5, 50 and 60 mg/m(2). The dose escalation was based on the dose-limiting toxicity (DLT) observed during the first cycle. RESULTS: Grade 4 neutropenia was observed at level 3, but no other DLT was observed at less than level 4 during the first cycle. However, three patients at level 3 could not continue treatment without a decrease in the dosage after the second cycle. Based on these results, level 2 was considered to be the clinically recommended dosages. CONCLUSION: Biweekly TXT and CPT-11 was well tolerated. The recommended dosages of TXT and CPT-11 for a phase II trial are 35 mg/m(2) and 50 mg/m(2), respectively. PMID- 14520529 TI - Neoadjuvant chemotherapy with cisplatin, 5-FU, and leucovorin (PLF) in locally advanced gastric cancer: a prospective phase II study. AB - BACKGROUND: Patients with locally advanced gastric cancer (cT3, cT4, N+, M0) have a dismal prognosis, despite complete resection. The objective of this study was to evaluate the toxicity and efficacy of neoadjuvant chemotherapy using the PLF (cisplatin/leucovorin [folinic acid]/5-fluorouracil [FU]) regimen in these patients. Primary endpoints of the study were the toxicity and the response to chemotherapy. Secondary endpoints were the rate of complete resection, survival, and first site of failure. METHODS: Forty-nine patients with adenocarcinoma of the stomach were enrolled. Staging was based on abdominal computed tomography (CT) scans, endosonography, and laparoscopy. The intention was to administer two cycles (each containing six courses) of preoperative chemotherapy, consisting of cisplatin 50 mg/m(2), high-dose folinic acid (HD-FA) 500 mg/m(2), and HD 5-FU (HD 5-FU) 2000 mg/m(2) (PLF). Following chemotherapy all patients were referred to surgery. To be evaluable for response, survival, and first site of failure, the patient had to receive at least one cycle of chemotherapy. RESULTS: Toxicity observed was low, with grade 3 toxicity in fewer than 5% of the patients and two events of grade 4 toxicity (diarrhea and pulmonary embolism). Forty-two of the patients (86%) received at least one cycle of chemotherapy. The clinical response rate in these patients was 26% (11/42 patients). In 76% of the patients (32/42), a complete resection was possible. The median duration of follow-up for the surviving patients was 58 months (range, 38 to 80+ months). The median survival time for the 42 patients assessable for response was 25.4 months (range, 6 to 80+ months). After complete resection, median survival time was 32 months (range, 7.6 to 80+ months). The median survival time for clinically responding patients has not yet been determined, but 5-year survival is 90%. Twenty of the 32 completely resected patients (62.5%) had recurrences. First site of failure was peritoneal dissemination in 10 patients; locoregional and distant recurrences were rare. CONCLUSION: Neoadjuvant chemotherapy with PLF in patients with locally advanced gastric cancer has low toxicity and reasonable efficacy, allowing administration on an outpatient basis. Clinically responding patients have an excellent outcome after complete resection. The development of peritoneal dissemination even after neoadjuvant chemotherapy and complete resection remains an unsolved problem in patients with nonintestinal type tumors. PMID- 14520530 TI - Evaluation of malignancy using Ki-67 labeling index for gastric stromal tumor. AB - BACKGROUND: Assessment of malignant potential in gastrointestinal stromal tumors (GISTs) is still problematic. The maximum tumor diameter and the mitotic index are generally used as an index of malignancy of GISTs. The Ki-67 labeling index has recently been used as an index of cell growth, and the prognosis of GISTs was reported to be significantly poor when the value of this index was 10% or higher. METHODS: Clinicopathological and immunohistological factors were analyzed in 15 patients who underwent surgical resection of gastric stromal tumors at our department between April 1997 and July 2002. The patients were divided into "metastasis/recurrence" and "benign" groups. Also, the relationship of changes in the Ki-67 labeling index to the degree of malignancy in recurrent lesions was assessed in an 84-year-old woman who underwent five reoperations because of recurrences in the peritoneum. RESULTS: Significant differences were noted between the metastasis/recurrence and benign groups in relation to the mean maximum tumor diameter (186.7 +/- 80.8 mm vs 41.3 +/- 22.9 mm), mitotic index (88.3 +/- 5.0/50 high-power fields [HPF] vs 3.0 +/- 2.9/50 HPF), and the Ki-67 labeling index (11.4 +/- 2.5% vs 0.01 +/- 0.51%). In the patient who had metastasis to the liver 3.5 years after initial operation and underwent five reoperations before death, the intervals until detection of recurrence tended to be shortened gradually. The Ki-67 labeling index varied with each operation, and tended to be higher at the time of reoperations than at the initial operation. CONCLUSION: The maximum tumor diameter, mitotic index, and Ki-67 labeling index were useful as an index of malignancy for gastric stromal tumor. The efficacy of surgical resection alone may be insufficient in patients with disseminated metastasis to the peritoneum. PMID- 14520531 TI - Detection of Epstein-Barr virus in gastric carcinoma cells and surrounding lymphocytes. AB - BACKGROUND: Epstein-Barr virus (EBV), the etiological agent of infectious mononucleosis, has an important role in the oncogenesis of EBV-related malignant diseases. The association of EBV with gastric carcinoma cells has become well known recently, but there are only a few reports concerning its association with surrounding epithelia and infiltrating lymphocytes. In this study, we investigated the association of EBV with gastric carcinoma and surrounding cells. METHODS: One hundred and two cases of gastric carcinoma were studied. The specimens were studied for the presence of the EBV genome by polymerase chain reaction (PCR), and then by in situ hybridization (ISH) technique to determine the localization of EBV. RESULTS: Of 97 informative cases, EBV was detected in 21 cases (21.6%) by the PCR method. ISH studies showed that EBV RNA was expressed in 5 of the 97 cases (5.2%) and was localized to the nuclei of carcinoma cells. All these 5 lesions were found in male patients. In these 5 cases, 3 were diffuse type and 2 were intestinal type, and all cases arose in the proximal region of the stomach. EBV RNA was not detected in non-neoplastic epithelia, but it was detected in 24 of the 97 cases (24.7%) in small lymphocytes. CONCLUSION: EBV was detected in 5.2% of gastric carcinomas and in 24.7% of infiltrating lymphocytes by the ISH method. The high positive rate (21.6%) by the PCR method corresponds to the presence of the EBV genome in surrounding lymphocytes. PMID- 14520532 TI - Intestinal phenotypic expression of gastric depressed adenomas and the surrounding mucosa. AB - BACKGROUND: Gastric depressed adenomas (DAs) have been reported to be more aggressive than nondepressed ones. To clarify any differences among them, we investigated the intestinal phenotypic expression of DAs and the surrounding mucosa. METHODS: Fifty DAs resected surgically or endoscopically were studied. Forty-two nondepressed adenomas (Non-DAs) were selected as controls. According to the classification of intestinal metaplasia, the phenotypes of the adenomas and surrounding mucosa were classified into four categories: complete intestinal type (I/Com-type), incomplete intestinal type (I/In-type), gastric type (G-type), and unclassified type (U-type), based on a combination of the expression of the antibodies CD10 (small-intestinal brush border), MUC2 (intestinal goblet cells) and HGM (gastric foveolar epithelium). RESULTS: I/In-type adenomas were frequently observed among DAs (60%), whereas I/Com-type adenomas were frequently seen in Non-DAs (52%). The incidence of phenotypes was significantly different among DAs and Non-DAs. Intestinal metaplasia was present in the surrounding mucosa of more than 90% of both DAs and Non-DAs. Surrounding mucosa of I/In-type was also frequently observed in DAs, as was that of I/Com-type in Non-DAs. The expression of p53 in DAs (8%) was higher than that in Non-DAs (5%). p53-positive adenomas were present only among the I/Com-type and I/In-type adenomas, while G type and U-type adenomas showed no p53 positivity. CONCLUSIONS: Both DAs and their surrounding mucosa appear to have an unstable status, because they possess a hybrid phenotype of I/In-type more commonly than Non-DAs. Such instability could be considered a risk factor for neoplastic potential and, therefore, DAs should be followed carefully. PMID- 14520534 TI - Simultaneous early adenocarcinoma and mucosa-associated lymphoid tissue (MALT) lymphoma of the stomach associated with Helicobacter pylori infection. AB - The simultaneous association of gastric carcinoma with gastric lymphoma is a rare event. Recent studies have suggested that not only gastric cancer but also primary gastric lymphomas, especially those of mucosa-associated lymphoid tissue (MALT) type, are associated with Helicobacter pylori infection. We report on a 51 year-old woman who was referred to our hospital for the evaluation of abnormal shadows revealed by an upper gastrointestinal radiography series. Endoscopy of the upper gastrointestinal tract revealed early cancer in the middle body of the stomach. Biopsy of the lesion subsequently proved it to be a signet-ring cell carcinoma. Total gastrectomy was performed, under a diagnosis of early gastric carcinoma. The resected specimen revealed two grossly separate lesions. Histological examination confirmed that the gastric body lesion was compatible with early moderately differentiated tubular adenocarcinoma of type 0-IIc, while the lesion of the fundus corresponded to MALT lymphoma. H. pylori was detected, and chronic gastritis was also present in the resected gastric specimen. H. pylori infection may have played a major role in the development of both the MALT lymphoma and the adenocarcinoma of the stomach in this patient. PMID- 14520533 TI - Complete response of a highly advanced gastric carcinoma to preoperative chemoradiotherapy with S-1 and low-dose cisplatin. AB - S-1 has been developed as a new oral anticancer drug, based on the biological modulation of 5-fluorouracil. We report a patient with highly advanced gastric carcinoma who was treated successfully with a new combination chemoradiotherapy using S-1 and cisplatin (CDDP). The patient was a 37-year-old man who was diagnosed with advanced gastric carcinoma (T4N3M0) that had invaded the diaphragm and the paraaortic tissues. Remarkable tumor reduction was observed in the primary tumor and metastatic lymph nodes around the stomach after three cycles of the therapy. Radiological examination before surgery determined that a partial response (PR) had been achieved by the initial therapy. Adverse effects included only a gastrointestinal disorder that was limited to grade 2 when low-dose CDDP was utilized in the regimen, while an initial high dose of CDDP resulted in grade 3 toxicity, due to myelosuppression. The patient underwent curative surgery, including total gastrectomy, D2 lymph node dissection, and splenectomy, after completion of the radiochemotherapy regimen. No surgical complication was observed. No tumor cells were detected by pathological evaluation of the resected stomach and all the regional lymph nodes, confirming a pathological complete response (CR; grade 3). This regimen is a potent treatment for advanced gastric carcinoma, especially when used as preoperative chemotherapy to control cancer cells. PMID- 14520535 TI - Recurrence in jejunal pouch after proximal gastrectomy for early upper gastric cancer. AB - Recently, jejunal pouch reconstruction after proximal gastrectomy has been a common procedure in Japan. However, according to our investigation, there are no reports of recurrence in the jejunal pouch. We experienced a patient with a recurrence in the jejunal pouch after proximal gastrectomy for early upper gastric cancer. In view of the pathological findings and the recurrence site, we suspected an implantation on the staple-line made for the jejunal pouch. PMID- 14520536 TI - Identification of bacteria recovered from clinical specimens by 16S rRNA gene sequencing. AB - The sequence of the 16S rRNA gene has been used extensively for phylogenetic classification, identification, and genotypic typing of bacteria. Identification of bacterial isolates by 16S rRNA gene sequencing, though generally performed in reference laboratories, has been recently introduced for routine use in clinical laboratories to identify isolates that cannot be identified by conventional methods. Described in this report is the use of 16S rRNA gene sequencing to identify uncommon bacteria, or bacteria with unusual phenotypic properties, with four brief case presentations to illustrate its clinical application. The feasibility, usefulness and limitations of performing this approach in the clinical laboratory are also discussed. PMID- 14520539 TI - Glioma invasion: mechanism, modulation and future possibilities. PMID- 14520537 TI - Na+-mediated piezoprotection in Rhodotorula rubra. AB - Sodium concentrations as low as 2 mM exerted a significant protective effect on the high-pressure inactivation (160-210 MPa) of Rhodotorula rubra at pH 6.5, but not on two other yeasts tested (Schizosaccharomyces pombe and Saccharomyces cerevisiae). A piezoprotective effect of similar magnitude was observed with Li+ (2 and 10 mM), and at elevated pH (8.0-9.0), but no effect was seen with K+, Ca2+, Mg2+, Mn2+, or NH4(+). Intracellular Na+ levels in cells exposed to low concentrations of Na+ or to pH 8.0-9.0 provided evidence for the involvement of a plasma membrane Na+/H+ antiporter and a correlation between intracellular Na+ levels and pressure resistance. The results support the hypothesis that moderate high pressure causes indirect cell death in R. rubra by inducing cytosolic acidification. PMID- 14520540 TI - The BrainIT group: concept and core dataset definition. AB - INTRODUCTION: An open collaborative international network has been established which aims to improve inter-centre standards for collection of high-resolution, neurointensive care data on patients with traumatic brain injury. The group is also working towards the creation of an open access, detailed and validated database that will be useful for post-hoc hypothesis testing. In Part A, the underlying concept, the group coordination structure, membership guidelines and database access and publication criteria are described. Secondly, in part B, we describe a set of meetings funded by the EEC that allowed us to define a "Core Dataset" and we present the results of a feasibility exercise for collection of this core dataset. METHODS: Four group meetings funded by the EEC have enabled definition of a "Core Dataset" to be collected from all centres regardless of specific project aim. A paper based pilot collection of data was conducted to determine the feasibility for collection of the core dataset. Specially designed forms to collect the core dataset demographic and clinical information as well as sample the time-series data elements were distributed by both email and standard mail to 22 BrainIT centres. A deadline of two months was set to receive completed forms back from centres. A pilot data collection of minute by minute physiological monitoring data was also performed. FINDINGS: A core-dataset was defined and can be downloaded from the BrainIT web-site (go to "Core dataset" link at: www.brainit.org). Eighteen centres (82%) returned completed forms by the set deadline. Overall the feasibility for collection of the core data elements was high with only 10 of the 64 questions (16%) showing missing data. Of those 10 fields with missing data, the average number of centres not responding was 12% and the median 6%. An SQL database to hold the data has been designed and is being tested. Software tools for collection of the core dataset have been developed. Ethics approval has been granted for collection of multi-centre data as part of a pilot data collection study. INTERPRETATION: The BrainIT network provides a more standardised and higher resolution data collection mechanism for research groups, organisations and the device industry to conduct multi-centre trials of new health care technology in patients with traumatic brain injury. PMID- 14520541 TI - Conservative (labyrinth-preserving) transpetrosal approach to the clivus and petroclival region--indications, complications, results and lessons learned. AB - OBJECTIVE: Tumours or vascular lesions of the clivus and juxtaclival region present a unique challenge to neurosurgeons and a variety of techniques, with a wide spectrum of complexity, have been advocated. This report presents the use of a conservative transpetrosal approach which combines partial removal of the postero-lateral petrous bone with preservation of the labyrinth, with particular focus on efficacy and the reduction of complications. PATIENTS AND METHODS: Over 9 years, 52 patients underwent a conservative combined supra-infratentorial, labyrinth-preserving transpetrosal approach. There were 32 women and 20 men; 33 had tumour lesions including 22 patients with a clival or petroclival meningioma. Among the 19 patients with a vascular lesion, 12 patients had a basilar artery trunk aneurysm, including one giant midbasilar aneurysm. 3 Patients had vertebro basilar junction aneurysms, and 4 patients had a pontine cavernoma. Data on the clinical features, investigations and operative techniques were extracted from the patient's case records. Outcome was assessed by serial examinations of the patients or by telephone interview with the treating physician, and in survivors graded as excellent, good or poor. RESULTS: 52 patients, 23 patients (44%) had an excellent outcome, in 21 patients (41%) the outcome was good and in 7 patients (13%) the outcome was poor. One patient with a complex fusiform basilar trunk aneurysm, operated upon as an emergency, died in the postoperative period. Total resection was achieved, as demonstrated by follow-up MRI in 20 of 33 patients with a tumour, including 15 of 22 patients with clival or petro-clival meningiomas. All vascular lesions were treated effectively and in 14 patients with a basilar or vertebro-basilar junction aneurysm, clipping of the aneurysm was achieved. In one patient, a giant calcified vertebro-basilar junction aneurysm was resected. New cranial nerve deficits or an accentuation of a pre existing deficit, occurred in 8 patients with a tumour and 4 patients with a vascular lesion. Complications included: temporary conductive hearing deficit in 4 patients, a minor laceration of the sigmoid/transverse sinus in 4 and transient post-operative CSF leakage in 12 patients. Temporary lumbar drainage resulted in sealing of the CSF leak in 8 patients, but in 3 patients a permanent ventriculo peritoneal shunt had to be implanted. CONCLUSION: When based on adequate experience in skull base surgery, the combined supra-infratentorial, conservative transpetrosal approach, with preservation of the labyrinth, allows direct and wide exposure of a large variety of tumour and vascular lesions located along the supra- and infratentorial juxta-clival area; little or almost no retraction of neurovascular structures is needed and with adherence to important principles, complications related to the approach can be minimised. PMID- 14520542 TI - Brain lesion size and phase shift as an index of cerebral autoregulation in patients with severe head injury. AB - BACKGROUND: Whether the phase relationship (phase shift) between cerebral blood flow velocity as assessed by transcranial Doppler ultrasound and blood pressure at 0.1 Hz can be used to assess cerebral autoregulation (CA) in patients with severe traumatic brain injury (TBI). METHODS: In 33 healthy volunteers (mean age, SD; 37+/-17 years, range 17-65) middle cerebral artery (MCA) blood velocity (V) was recorded simultaneously with finger blood pressure (BP) over a period of 10 minutes under normocapnic and hypocapnic conditions to generate normative data. In 27 patients with severe TBI (Glasgow Coma scale score < or =8) serial close in time investigations of cranial computed tomography (CT) scanning and phase shift assessment were performed on days 1, 3, 5, and 8 after trauma. Phase shift in the MCA was compared to brain parenchyma lesion size in the MCA territory on CT scanning. Lesion size was classified into 0, normal; 1, presence of a small lesion (diameter <3 cm); 2, presence of a large lesion (>3 cm). FINDINGS: Compared to normocapnia, hypocapnia significantly increased phase shift at 0.1 Hz from 78+/-28 degrees to 101+/-25 degrees (p < 0.001). In the TBI patients, 115 comparisons between CT findings and CA results were possible. Phase shift detected a pathological CA in 31 instances, which were more frequent in CT lesion type 2 (19/42) than in group 0 (7/44) and group 1 (5/29). INTERPRETATION: When CA is intended to be assessed by use of phase shift, the hyperventilation setting needs its own reference values. In MCA territories containing a traumatic lesion greater than 3 cm in diameter phase shift at 0.1 Hz will detect a high frequency (44%) of a disturbed state of CA. PMID- 14520543 TI - Association between apolipoprotein E genotype and outcome of traumatic brain injury. AB - BACKGROUND: The prognosis of traumatic brain injury is quite variable and not fully explained by the known factors. This study is to examine if the polymorphism of apolipoprotein E (apoE) influences the outcome of traumatic brain injury. METHODS: Over a period of twelve months, we prospectively studied 100 patients who sustained traumatic brain injuries and were admitted to our neurosurgical unit. FINDINGS: Nineteen patients were apoE4+ and 81 patients were apoE4-. There was no significant difference between apoE4+ and apoE4- groups in the cause of injury (p=0.288), type of brain injury (p=0.983) and choice of treatment (p=0.88). The proportion of patients with a lower GCS (<13), indicating a poor prognosis, was higher in the apoE4+ group (73.7%) than that in apoE4- group (61.7%), although the difference was not significant (p=0.654). Six patients (7.4%) in the apoE4- group and 5(26.3%) in the apoE4+ group had been drinking alcohol at the time of injury (p=0.018). The mean duration of hospital stay for apoE4+ patients was significantly longer than for apoE4- patients (p<0.001). Six months after injury, 10 of 19 patients (52.6%) with apoE4 had an unfavorable outcome (dead, vegetative state, or severe disability) compared with 20 of the 81 (24.1%) patients without apoE4 (p=0.017). The apoE4+ patients had a significantly longer hospital stay and unfavorable outcomes after brain injury. INTERPRETATION: This study discloses a significant genetic association between the apoE genotypes and outcomes of traumatic brain injury. Patients with apoE4 allele are more likely to have an unfavorable clinical outcome after brain injury. PMID- 14520544 TI - Value of the quantity and distribution of subarachnoid haemorrhage on CT in the localization of a ruptured cerebral aneurysm. AB - BACKGROUND: Computed tomography (CT) is the "gold standard" for detecting subarachnoid haemorrhage (SAH) and digital subtraction angiography (DSA) for visualising the vascular pathology. We studied retrospectively 180 patients with subarachnoid haemorrhage (SAH) who underwent first non-enhanced computed tomography (CT), then digital subtraction angiography (DSA) and finally operative aneurysm clipping. Our aim was to assess if the location of the ruptured aneurysm could be predicted on the basis of the quantity and distribution of haemorrhage on the initial CT scan. METHODS: 180 patients with SAH were retrospectively studied. All the CT and DSA examinations were performed at the same hospital. CT was performed within 24 hours after the initial haemorrhage. DSA was performed after the CT, within 48 hours after the initial haemorrhage. Two neuroradiologists, blind to the DSA results, analysed and scored independently the quantity and distribution of the haemorrhage and predicted the site of the ruptured aneurysm on the basis of the non-enhanced CT. DSA provided the location of the ruptured aneurysm. All the patients were operated upon, and the location of the ruptured aneurysm was determined. FINDINGS: The overall reliability value (kappa-value) between the two neuroradiologists for locating all ruptured aneurysms was 0.780. The corresponding value for the right MCA was 0.911, that for the left MCA 0.877 and that for the AcoA 0.736. Not all of the kappa-values were calculated, either because the location of the rupture was constant or because the number of ruptures in the vessel was too small. Subarachnoid haemorrhage with a parenchymal hematoma is an excellent predictor of the site of the ruptured aneurysm with a statistical significance of p=0.003. INTERPRETATION: The quantity and pattern of the blood clot on CT within the day of onset of SAH is a reliable and quick tool for locating a ruptured MCA or AcoA aneurysm. It is not, however, reliable for locating other ruptured aneurysms. Subarachnoid haemorrhage with a parenchymal hematoma is an excellent predictor of the site of a ruptured aneurysm. PMID- 14520545 TI - Importance of re-examination for medulloblastoma and atypical teratoid/rhabdoid tumor. AB - Medulloblastoma may can be difficult to distinguish from atypical teratoid/rhabdoid tumor (AT/RT), since they resemble each other histologically. We re-examined whether AT/RT was included among cases who had been diagnosed as medulloblastoma. All of fifteen medulloblastomas (10 males and 5 females) diagnosed at the Kitasato University Hospital were collected and stained immunohistochemically. Two cases originally diagnosed as medulloblastoma were reclassified as AT/RT based on histological re-examination including immunohistochemical studies. While these two cases of AT/RT were found during infancy, only one medulloblastoma was found in infancy.Histologically, small rhabdoid cells and large, pale, bland cells were common but typical rhabdoid cells were not seen in the two AT/RTs. Gland-like structures were also seen. The tumor cells in AT/RT, but not those in medulloblastoma, were immunoreactive for vimentin, epithelial membrane antigen and smooth muscle actin. In conclusion, if a diagnosis of medulloblastoma is made histologically, it should be confirmed immunohistologically, since it is difficult to distinguish AT/RT from medulloblastoma. When appropriate treatment was specifically targeted at AT/RT it may improve the outcome. PMID- 14520546 TI - C-reactive protein levels following standard neurosurgical procedures. AB - BACKGROUND: The aim of the present study was to establish the magnitude and time course of C-reactive protein increases following routine neurosurgical procedures in the absence of clinical and laboratory signs of infection. METHOD: C-reactive protein levels were studied daily following ventriculo-peritoneal shunt implantation, anterior cervical fusion, vestibular schwannoma operation, supratentorial glioma surgery, endovascular intracranial aneurysm treatment and open cerebral aneurysm surgery. FINDINGS: The magnitude of the C-reactive protein increase depended on the extent of surgical trauma and peak-levels were recorded between postoperative day one and four after which the levels tapered off. INTERPRETATION: Increases occurring after the fourth postoperative day are likely to be caused by complications of surgery, e.g. infection. PMID- 14520547 TI - The attenuation of vasospasm by using a sod mimetic after experimental subarachnoidal haemorrhage in rats. AB - BACKGROUND: Delayed cerebral vasoconstriction and brain ischemia, are critical problems in the management of a patient affected by rupture of an intracranial aneurysm. Overexpression of Cu-Zn superoxide dismutase (Cu-Zn SOD) can reduce the extent of cerebral vasospasm. We, therefore investigated if vasospasm, can be prevented by a novel, stable, and cell permeable SOD mimetic, MnTBAP [Mn(III) tetrakis (4-benzoic acid) porphyrin] which permeates the biological membranes and scavenges superoxide anions and peroxynitrite. METHODS: 28 rats (225-250 g) were divided equally into four groups: group 1: control; group 2: SAH only; group 3: SAH plus placebo; and group 4: SAH plus MnTBAP. We used a double haemorrhage method to produce SAH. Starting six hours after SAH, 5 mg/kg MnTBAP (Calbiochem, Darmstadt-Germany; Cat. No 475870)) or an equal volume of 0.9% saline (37 degrees C) was administered by intraperitoneal injection twice daily for 5 days to groups 4 and 3 respectively. MnTBAP or 0.9% saline injections were continued up to fifth day after SAH and rats were sacrificed on the fifth day. Brain sections at the level of the pons were examined by light microscopy. Planimetric measurements were made for the cross-sectional areas of the lumen and the vessel wall (intima plus media) of the basilar artery by a micrometer. FINDING: Administration of MnTBAP significantly attenuated the vasoconstriction of the basilar artery in group 4 compared with the groups 2 and 3 (p<0.001). INTERPRETATION: These results suggest that this SOD mimetic (MnTBAP) attenuates delayed cerebral vasoconstriction following experimental SAH and that superoxide anions have a role in the pathogenesis of vasospasm after SAH. PMID- 14520548 TI - Outcome assignment in the International Surgical Trial of Intracerebral Haemorrhage. PMID- 14520549 TI - The potential for strategies using micronutrients and heterocyclic drugs to treat invasive gliomas. AB - Local invasion of neoplastic cells into the surrounding brain is perhaps the most important aspect of the biology of gliomas that precludes successful therapy. Despite significant advances in neuro-imaging, neurosurgery and radiotherapy, the median survival for patients with a malignant glioma is still less than one year. With the increasing knowledge of the biology of brain tumours, derived from cellular and molecular studies, new methods of treatment are being developed with some success. Approaches studied already include anti-invasive, pro-apoptotic and anti-angiogenesis strategies and clinical trials are imminent. In this article we review two new approaches to the management of gliomas: nutraceutical intervention and heterocyclic drugs. The first approach uses a combination of naturally occurring agents, including citrus flavonoids, chokeberry extract, red grape seed extract, lycopene, selenium and red clover extract. These agents can either trigger apoptosis or affect the pathways underlying diffuse invasion. The second approach involves the use of a heterocyclic drug, clomipramine, which selectively triggers apoptosis in neoplastic cells but not in normal glia. The article refers to the results of recent studies performed in our laboratory which suggest that these new approaches can be translated into benefit to patients. PMID- 14520550 TI - Pertreatment radiation induced oedema causing acute hydrocephalus after radiosurgery for multiple cerebellar metastases. AB - BACKGROUND: Most complications of radiosurgery occur after 6-12 months and some long term complications up to after 5-10 years. Although severe, immediate side effects are rare, we report a patient who shows that this possibility should be taken into account when large volumes are treated especially in the posterior fossa. CASE REPORT: A patient was treated with radiosurgery for multiple brain metastases, of which 5 were located in the posterior fossa. Deterioration occurred, radiosurgery was halted abortion and CT showed acute hydrocephalus caused by a significant edema in the posterior fossa, not present in an MR examination performed some 6 hours earlier. After intubation and external ventricular CSF drainage, a permanent shunt was inserted, the edema resolved, and the patient recovered to her pretreatment condition within 10 days. The remaining metastases were treated during the first session and MR images showed the hydrocephalus resolved when all the radiated tumors had significantly decreased in size. INTERPRETATION: Care should be taken when treating with radiosurgery patients posterior fossa metastases and signs of compressed CSF pathways. Exceptionally, an acute radiation induced edema could result in a block of the CSF pathways, necessitating a surgical intervention. Prophylactic shunt insertion or hospitalization the night following the treatment should be considered. PMID- 14520551 TI - Stereotactic radiofrequency ablation for sessile hypothalamic hamartoma with an image fusion technique. AB - BACKGROUND: Radiosurgery has been advocated as a primary treatment for hypothalamic hamartoma (HH), but it has a risk of damaging the surrounding structures and does not have an immediate effect for refractory epilepsy, endocrinological and mental disorders. METHOD: We report on a 13-year-old boy with a large and sessile HH who presented with intractable seizures, precocious puberty and aggressiveness. Stereotactic radiofrequency ablation (SRA) combined with an image fusion technique was performed to make a maximum ablative lesion within the HH via multiple trajectories. FINDINGS: After surgery, we observed rapid cessation of the gelastic seizures and aggressiveness. The ophthalmological function did not get worse, and the hypothalamopituitary function improved. INTERPRETATION: SRA in combination with an image fusion technique is a viable alternative treatment for HH, because it provides precise preoperative simulation and immediate improvement of symptoms can be obtained. PMID- 14520552 TI - Intracerebral abscess formation following embolisation of an internal carotid artery aneurysm using Guglielmi detachable coils. AB - Infection following endovascular therapy for cerebrovascular disease is a potential but rare complication. A 70-year-old lady in whom an intracranial abscess formed secondary to GDC embolisation of a giant right internal carotid artery aneurysm is reported. Computed tomography (CT) showed the abscess and staphylococcus aureus was cultured from the cerebrospinal fluid and blood. The abscess was successfully treated by antibiotic therapy. Infected Guglielmi detachable coils (GDC) may result in abscess formation in the presence of underlying cerebral ischaemia. PMID- 14520553 TI - Syringomyelia and the arachnoid web. AB - Three cases of syringomyelia associated with arachnoid webs are reported. Each patient presented with progressive myelopathy and had thoracic syringes detected on magnetic resonance imaging (MRI). In one patient the web was also visible. At operation a thoracic arachnoid web was found, obstructing the subarachnoid compartment in each patient. One patient had intraoperative ultrasound, which demonstrated caudal web movement with each cardiac systole. The webs were divided and shunts inserted into the syringes. All patients improved clinically, and on follow-up MRI. Arachnoid webs are likely to represent a focal band of arachnoiditis and are difficult to visualise on standard preoperative MR imaging. A reduction in the subarachnoid space compliance with resultant increase in pulse pressure and potentiation of an arterial pulsation driven perivascular flow could explain the associated syringes. Treatment should be aimed at restoring compliance, and involve division of the web with or without shunt insertion. PMID- 14520554 TI - Extensive craniofacial subcutaneous emphysema after evacuation of a chronic subdural haematoma. AB - An unusual complication of craniofacial subcutaneous emphysema after evacuation of a chronic subdural haematoma is reported. The emphysema caused massive swelling and deformity of the patient's head and face, as well as the occlusion of both eyes, and it resolved spontaneously. The possible mechanisms of development of emphysema are discussed. PMID- 14520555 TI - Ultra rapid spontaneous resolution of acute posttraumatic subdural hematomas in patient with temporal linear fracture. AB - We report a case of 57 year-old man with documented posttraumatic acute subdural hematoma and a linear temporal bone fracture. He suffered from a blunt head injury and presented with sudden loss of consciousness. Within 2 hours he became alert and oriented. Follow-up CT scan of brain 2 hours after the initial one showed resolution and redistribution of the subdural hematoma. To our knowledge, this is the first case in the literature about spontaneous resolution of an acute subdural hematoma in a patient with a linear fracture and the fastest resolution period. In this article, the authors discuss the underlying pathophysiology of this uncommon phenomenon. PMID- 14520556 TI - Sinus thrombosis probably resulting from a dural arteriovenous fistula development in the superior sagittal sinus. PMID- 14520557 TI - An autopsy case of giant aneurysm of vertebrobasilar artery treated by endovascular surgery. PMID- 14520558 TI - The 22nd Annual Working Group in Interventional Neuroradiology--Val D'Isere, France, January 14th-17th 2003. PMID- 14520559 TI - Camping programs for children with cancer and their families. AB - This paper presents an introduction to the role of camping programs for children with cancer and their families and a hypothesized theory of action. Nine previously published studies investigating recreation-based summer camps for children with cancer and their families are summarized. Overall, these programs appear to be effective; however, an overall conclusion is difficult to state due to variations in design, heterogeneity of subjects, and differences in the definition of camp programs. The lack of knowledge regarding the efficacy of these programs is identified as the largest deficiency in the current literature on this topic. Future research needs in this area are discussed. PMID- 14520560 TI - Evidence for a modifying pathway in SMA discordant families: reduced SMN level decreases the amount of its interacting partners and Htra2-beta1. AB - Proximal spinal muscular atrophy (SMA) is a neuromuscular disorder caused by homozygous mutations of the SMN1 gene. SMN1 interacts with multiple proteins with functions in snRNP biogenesis, pre-mRNA splicing and presumably neural transport. SMN2, a nearly identical copy of SMN1, produces predominantly exon 7-skipped transcripts, whereas SMN1 mainly produces full-length transcripts. The SR-like splicing factor Htra2-beta1 facilitates correct splicing of SMN2 exon 7 through direct interaction with an exonic splicing enhancer within exon 7. In rare cases, siblings with identical 5q13-homologues and homozygous absence of SMN1 show variable phenotypes, suggesting that SMA is modified by other factors. By analysing nine SMA discordant families, we demonstrate that in all families unaffected siblings produce significantly higher amounts of SMN, Gemin2, Gemin3, ZPR1 and hnRNP-Q protein in lymphoblastoid cell lines, but not in primary fibroblasts, compared with their affected siblings. Protein p53, an additional SMN-interacting protein, is not subject to an SMN-dependent regulation. Surprisingly, Htra2-beta1 is also regulated by this tissue-specific mechanism. A similar regulation was found in all type I-III SMA patients, although at a different protein level than in discordant families. Thus, our data show that reduced SMN protein levels cause a reduction in the amount of its interacting proteins and of Htra2-beta1 in both discordant and non-discordant SMA families. We provide evidence that an intrinsic SMA modifying factor acts directly on the expression of SMN, thus influencing the SMA phenotype. Further insights into the molecular pathway and the identification of SMA modifying gene(s) may help to find additional targets for a therapy approach. PMID- 14520561 TI - Effects of Ca(2+) channel blockers and protein kinase/phosphatase inhibitors on growth and anthraquinone production in Rubia cordifolia callus cultures transformed by the rolB and rolC genes. AB - The transformation of Rubia cordifolia L. cells by the 35S- rolB and 35S- rolC genes of Agrobacterium rhizogenes caused a growth inhibition of the resulting cultures and an induction of the biosynthesis of anthraquinone-type phytoalexins. Inhibitor studies revealed a striking difference between the rolC- and rolB-gene transformed cultures in their sensitivity to verapamil, an L-type Ca(2+) channel blocker. The rolC culture possessed a 2-fold lowered resistance to the inhibitor than the normal culture, while the rolB culture was 4-fold more resistant to the treatment. Additionally, growth of the rolC culture was totally inhibited when the culture was grown in Ca(2+)-free medium, whereas growth of the rolB culture was reduced by less than half. We interpreted these results as evidence for a lack of calcium homeostasis in both transgenic cultures. Anthraquinone (AQ) production was not inhibited in the normal or transformed cultures by the Ca(2+) channel blockers verapamil and LaCl(3), or by diphenylene iodonium, an inhibitor of NADPH oxidase, or by the protein kinase inhibitor staurosporine. These results indicate that the induction of AQ production in non-transgenic and transgenic cultures does not proceed through the activation of the common Ca(2+)-dependent NADPH oxidase pathway that mediates signal transduction between an elicitor receptor complex via transcriptional activation of defense genes. Okadaic acid and cantharidin, inhibitors of protein phosphatases 1 and 2A, caused an increase in AQ production in transgenic cultures. Okadaic acid stimulated AQ accumulation in the non-transformed culture, whereas cantharidin had no effect. These results show that different phosphatases are involved in AQ synthesis in normal and transgenic cultures of R. cordifolia. PMID- 14520563 TI - Generation of cyanogen-free transgenic cassava. AB - Cassava ( Manihot esculenta Crantz.) is the major source of calories for subsistence farmers in sub-Saharan Africa. Cassava, however, contains potentially toxic levels of the cyanogenic glucoside, linamarin. The cyanogen content of cassava foods can be reduced to safe levels by maceration, soaking, rinsing and baking; however, short-cut processing techniques can yield toxic food products. Our objective was to eliminate cyanogens from cassava so as to eliminate the need for food processing. To achieve this goal we generated transgenic acyanogenic cassava plants in which the expression of the cytochrome P450 genes ( CYP79D1 and CYP79D2), that catalyze the first-dedicated step in linamarin synthesis, was inhibited. Using a leaf-specific promoter to drive the antisense expression of the CYP79D1/ CYP79D2 genes we observed up to a 94% reduction in leaf linamarin content associated with an inhibition of CYP79D1 and CYP79D2 expression. Importantly, the linamarin content of roots also was reduced by 99% in transgenic plants having between 60 and 94% reduction in leaf linamarin content. Analysis of CYP79D1/ CYP79D2 transcript levels in transgenic roots indicated they were unchanged relative to wild-type plants. These results suggest that linamarin is transported from leaves to roots and that a threshold level of leaf linamarin production is required for transport. PMID- 14520562 TI - The unusual Arabidopsis extensin gene atExt1 is expressed throughout plant development and is induced by a variety of biotic and abiotic stresses. AB - We detail the expression of the Arabidopsis thaliana (L.) Heynh. atExt1 extensin gene. atExt1 is normally expressed in roots and inflorescences, and is induced by wounding, exogenously supplied salicylic acid, methyl jasmonate, auxins and brassinosteroids. Northern assays and histochemical analysis of transgenics expressing an atExt1:: gus fusion show that this gene is also induced by the brassica pathogen Xanthomonas campestris pv. campestris and that this induction is restricted to tissues close to the site of infection. Expression at regions of abscission and senescence also implicates atExt1 in these important developmental processes. PMID- 14520564 TI - Sugarbeet (Beta vulgaris L.): shoot regeneration from callus and callus protoplasts. AB - The successful application of recombinant DNA technology for crop plants requires efficient regeneration systems. A detailed study on the regeneration potential of callus and callus-derived protoplasts of a recalcitrant species, sugarbeet, was performed. A reproducible and highly efficient method for induction of regenerable friable callus was established from etiolated hypocotyl explants. A reduced sucrose concentration proved beneficial. Successful shoot regeneration could be demonstrated in 10 out of 12 tested lines. Seed germination, followed by callus induction and shoot regeneration required only a single culture medium. Additionally, the regeneration capacity of roots and root-derived callus was demonstrated. Highly efficient plant regeneration was also achieved when using protoplasts isolated from regenerable friable callus induced on etiolated hypocotyls explants. To our knowledge this represents the first report on callus protoplast to plant regeneration in sugarbeet. PMID- 14520565 TI - Type-h thioredoxins accumulate in the nucleus of developing wheat seed tissues suffering oxidative stress. AB - Thioredoxin h (Trx h) proteins are ubiquitous in all wheat organs, but show the highest accumulation in mature seeds. This distribution suggests the expression of Trx h during seed development. In the present study, we have analyzed the pattern of Trx h expression in developing wheat ( Triticum aestivum L.) seeds. Northern blot analysis detected a single band at any stage of development, which corresponded to the expression of at least two genes, TrxhA and TrxhB, as shown by competitive reverse transcription-polymerase chain reaction experiments. The analysis of the content of Trx h polypeptides showed the highest content in the embryo. The spatial pattern of accumulation of these proteins was established by immunocytological techniques. At early stages of development Trx h proteins localized to maternal tissues (nucellus projection cells and pedicel), the route of transport of nutrients to the developing endosperm. In the endosperm, Trx h proteins accumulated at a high level in the aleurone layer. At later stages of development, during seed maturation, Trx h proteins localized predominantly to the nucleus of both aleurone and scutellum cells, a feature exclusive of these seed tissues. The nuclear localization of Trx h proteins was associated with oxidative stress in these tissues, as shown by in situ staining of superoxide radicals in developing and germinating seeds. PMID- 14520566 TI - Discovery of an extended bundle sheath in Ricinus communis L. and its role as a temporal storage compartment for the iron chelator nicotianamine. AB - The extended bundle sheath (EBS) is a specialized layer of cells that enhances the lateral transport of photoassimilates within the leaf. This little-known tissue is often considered to be legume-specific. We identified an EBS in cotyledons and leaves of the non-legume Ricinus communis L. By means of cytological and immunological studies and using the localization of the iron chelator nicotianamine as an established indicator for mass transport, we confirmed its role as a transport tissue and a temporal sink. Observations on cotyledons of Ricinus seedlings further proved that the EBS carries out these tasks from a very early stage of development onwards. This is the first time that information has been obtained on the physiological role of an EBS in a non legume. Our results support the idea of its widespread occurrence among higher plants. PMID- 14520567 TI - The involvement of Ca(2+) gradients, Ca(2+) fluxes, and CaM kinase II in polarization and germination of Silvetia compressa zygotes. AB - Previous work has shown that distinct Ca(2+) gradients precede and predict the loci of germination of the zygotes of the brown alga, Silvetia compressa (J. Agardh) E. Serrao, T.O. Cho, S.M. Boo et Brawley, that are polarized by unilateral blue light. We show here that dark-grown S. compressa zygotes also form cytosolic Ca(2+) gradients prior to germination and then germinate from the site of elevated Ca(2+). In no case did germination occur without a prior formation of a Ca(2+) gradient. Using the self-referencing Ca(2+)-selective probe, we measured highly localized influx of Ca(2+) during photopolarization, indicating that extracellular stores supply at least some of the Ca(2+) needed to construct a gradient. Finally, we find that germination was inhibited by a bath applied inhibitor of calcium/calmodulin-dependent kinase II (CaM kinase II), KN 93 (but not by its inactive analog, KN-92), and by an injected inhibitory peptide for the kinase. KN-93 did not interfere with the photopolarization of the zygotes, consistent with the view that calmodulin is not involved in the initial response to light. The KN-93 results indicate that the requirement for active CaM kinase II for germination ends about 2 h before overt germination. We conclude that Ca(2+) gradients, generated in part by localized calcium entry from the seawater, are an essential part of the process of polarity development and expression in these cells, regardless of the nature of the external cue that directs the orientation of the axis. Calmodulin and CaM kinase II are involved in interpreting (but not in establishing) the calcium gradient, allowing germination to occur at the site of elevated calcium, but CaM kinase II appears not to be involved in the initiation of germination. PMID- 14520569 TI - Extensibility of isolated cell walls in the giant tip-growing cells of the xanthophycean alga Vaucheria terrestris. AB - Apical cell wall fragments isolated from the giant-cellular xanthophycean alga Vaucheria terrestris sensu Gotz were inflated with silicone oil by applying internal pressure ranging from 0.1 to 0.7 MPa, and the time-course of cell wall deformation was recorded and analyzed by videomicroscopy. Cell wall extensibility in the tip-growing region was estimated by the pressure required for cell wall extension, the amount of total extension until cell wall rupture and the rate of cell wall extension. Apical cell walls exhibited gradual extension, or creep, during inflation, which was eventually followed by rupture at the apical portion, whereas no appreciable extension was found in the cylindrical basal portion of the cell wall fragment. Besides the largest extension observed around the tip, substantial extension was also observed along the subapical region of the cell wall. The wall extensibility was dependent on the buffer pH used for infiltration before inflation. The optimum pH for the extension was about 8.0, but the cell wall was much less extensible after infiltration with an acidic buffer. Cell wall extensibility was dependent on the pH of the buffer used before inflation, regardless of that used in the previous infiltration. Moreover, pretreatment of the cell wall with a protease caused considerable loosening of cell walls, but affected the pH dependence of cell wall extensibility little. These results indicate that the extensibility of the cell walls in the giant tip-growing cells of the alga is distinct from that of plant cells that exhibit "acid growth" in its dependence on environmental pH and the role of cell wall proteins. PMID- 14520568 TI - Lithium treatment induces a hypersensitive-like response in tobacco. AB - Treatment of tobacco ( Nicotiana tabacum L.) plants with lithium induces the formation of necrotic lesions and leaf curling as in the case of incompatible pathogen interactions. Further similarities at the molecular level include accumulation of ethylene and of salicylic and gentisic acids, and induced expression of pathogenesis-related PR-P, PR5 and PR1 genes. With the exception of PR1 induction, lithium produced the same effects in transgenic tobacco plants that do not accumulate salicylate because of overexpression of the bacterial hydroxylase gene nahG. On the other hand, inhibition of ethylene biosynthesis with aminoethoxyvinylglycine prevented lithium-induced cell death and PR5 expression. These results suggest that lithium triggers a hypersensitive-like response where ethylene signalling is essential. PMID- 14520570 TI - Repeated freeze-thaw cycles induce embolism in drought stressed conifers (Norway spruce, stone pine). AB - Freezing and thawing lead to xylem embolism when gas bubbles caused by ice formation expand during the thaw process. However, previous experimental studies indicated that conifers are resistant to freezing-induced embolism, unless xylem pressure becomes very negative during the freezing. In this study, we show that conifers experienced freezing-induced embolism when exposed to repeated freeze thaw cycles and simultaneously to drought. Simulating conditions at the alpine timberline (128 days with freeze-thaw events and thawing rates of up to 9.5 K h( 1) in the xylem of exposed twigs during winter), young trees of Norway spruce [Picea abies (L.) Karst.] and stone pine (Pinus cembra L.) were exposed to 50 and 100 freeze-thaw cycles. This treatment caused a significant increase in embolism rates in drought-stressed samples. Upon 100 freeze-thaw cycles, vulnerability thresholds (50% loss of conductivity) were shifted 1.8 MPa (Norway spruce) and 0.8 MPa (stone pine) towards less negative water potentials. The results demonstrate that freeze-thaw cycles are a possible reason for winter-embolism in conifers observed in several field studies. Freezing-induced embolism may contribute to the altitudinal limits of conifers. PMID- 14520571 TI - Regulation of gibberellin 20-oxidase gene expression and gibberellin content in citrus by temperature and citrus exocortis viroid. AB - A cDNA clone coding for a gibberellin (GA) 20-oxidase ( CcGA20ox1), an enzyme of GA biosynthesis, which when expressed in vitro catalyzed the conversion of GA(12) to GA(9) and of GA(53) to GA(20), was isolated from the citrus hybrid Carrizo citrange (C itrus sinensis x Poncirus trifoliata). Transcripts of CcGA20ox1 were abundant in the apex and leaves and much less abundant in internodes, nodes and roots. Seedlings of Carrizo citrange cultured under a 32 degrees C/27 degrees C (day/night) regime elongated more than seedlings growing under 17 degrees C/12 degrees C conditions. The effect of higher temperature was associated with more CcGA20ox1 transcripts and with higher content of GA(1), the main active GA in citrus, in the shoot. The infection of Etrog citron ( Citrus medica) plants with citrus exocortis viroid (CEVd), which produces a stunted phenotype, reduced the levels of transcripts in the apical shoot hybridizing to the gene CcGA20ox1 of Carrizo citrange and the content of GA(1). Thus GA(1) content correlated with CcGA20ox1 transcript levels. In contrast, results for gibberellic acid (GA(3)) and paclobutrazol applications to Carrizo citrange showed that CcGA20ox1 expression was subject to feed-back regulation. These observations indicate that the feed-back regulation of GA20ox operates mostly when the levels of active GAs have been dramatically altered. The results also show that the growth reduction induced by environmental (temperature) and biotic (CEVd) factors may be partially due to the modulation of the expression of GA20ox genes. PMID- 14520572 TI - Relative turnover numbers of maize endosperm and potato tuber ADP-glucose pyrophosphorylases in the absence and presence of 3-phosphoglyceric acid. AB - Adenosine diphosphate glucose pyrophosphorylase (AGPase; EC 2.7.7.27) synthesizes the starch precursor, ADP-glucose. It is a rate-limiting enzyme in starch biosynthesis and its activation by 3-phosphoglyceric acid (3PGA) and/or inhibition by inorganic phosphate (Pi) are believed to be physiologically important. Leaf, tuber and cereal embryo AGPases are highly sensitive to these effectors, whereas endosperm AGPases are much less responsive. Two hypotheses can explain the 3PGA activation differences. Compared to leaf AGPases, endosperm AGPases (i) lack the marked ability to be activated by 3PGA or (ii) they are less dependent on 3PGA for activity. The absence of purified preparations has heretofore negated answering this question. To resolve this issue, heterotetrameric maize ( Zea mays L.) endosperm and potato ( Solanum tuberosum L.) tuber AGPases expressed in Escherichia coli were isolated and the relative amounts of enzyme protein were measured by reaction to antibodies against a motif resident in both small subunits. Resulting reaction rates of both AGPases are comparable in the presence but not in the absence of 3PGA when expressed on an active-protein basis. We also placed the potato tuber UpReg1 mutation into the maize AGPase. This mutation greatly enhances 3PGA sensitivity of the potato AGPase but it has little effect on the maize AGPase. Thirdly, lysines known to bind 3PGA in potato tuber AGPase, but missing from the maize endosperm AGPase, were introduced into the maize enzyme. These had minimal effect on maize endosperm activity. In conclusion, the maize endosperm AGPase is not nearly as dependent on 3PGA for activity as is the potato tuber AGPase. PMID- 14520573 TI - Plant-specific regulation of replication protein A2 (OsRPA2) from rice during the cell cycle and in response to ultraviolet light exposure. AB - DNA replication is a process that is highly conserved among eukaryotes. Nonetheless, little is known about the proteins involved in it in plants. Replication protein A (RPA) is a heterotrimeric, single-stranded DNA-binding protein with several functions in DNA metabolism in humans and yeast and supposedly also in plants. Here we report on the regulation of OsRPA2, the 32-kDa subunit of RPA from rice ( Oryza sativa L.). We found conserved regulation mechanisms at the level of gene expression between animal and plant RPA2 genes and distinct features of OsRPA2 regulation at the level of protein expression. Unlike in animals or in yeast, protein abundance in rice was regulated in a cell cycle phase-specific manner and was altered after UV-C light exposure. On the other hand, posttranslational modification through phosphorylation did not appear to play a pivotal role in rice as it does in animal cells. Our results indicate that plant-specific mechanisms of regulation have evolved for RPA2 within the generally well-conserved process of DNA replication, suggesting specific requirements for regulation of DNA metabolism in plants as compared to other eukaryotes. PMID- 14520574 TI - A gene encoding a germin-like protein, identified by a cDNA-AFLP approach, is specifically expressed during germination of Phaseolus vulgaris. AB - In order to identify markers of germination in Phaseolus vulgaris L., a cDNA amplified fragment length polymorphism (AFLP) approach was conducted on mRNAs from embryo axes and from cotyledons. Among changes observed throughout the germination process, a cDNA fragment not detected 9 h after imbibition (HAI) but present specifically in axes 24 HAI was further studied. The complete cDNA was recovered by rapid amplification of cDNA ends, then cloned and sequenced. It includes an open reading frame predicting a 206-amino-acid polypeptide of 21.8 kDa. Analysis of the nucleotide sequence and deduced amino acid sequence revealed a high homology with germin-like proteins (GLPs), and particularly with an auxin binding protein from peach, ABP19, that belongs to the GLP family. Thus, we propose that this cDNA encodes the first GLP described in P. vulgaris, designated PvGLP1. Northern blot analysis carried out on mRNAs from seed axes showed a dramatic increase in PvGLP1 expression a few hours before radicle emergence (17 HAI). Among mature vegetative tissues, PvGLP1 expression was very weak in pods and not detected in leaves, stems or roots. Immunoblot analysis using antibodies raised against AtGER3 from Arabidopsis thaliana showed that the protein could be detected only in axes from the dry seed stage onwards, at a steady-state level. Then, PvGLP1 expression seems to be associated with the early stages of embryo axis growth. The high homology indicated with ABP19 led us to study the effect of different concentrations of indole-3-acetic acid (IAA) on PvGLP1 expression during germination. Whereas no effect was noticed at low concentrations (1, 5, 10 microM), a marked decrease in PvGLP1 mRNA level was observed in axes of seeds imbibed with 100 microM IAA. Thus, PvGLP1 gene expression is not stimulated by auxin and, moreover, it might be inhibited by high concentrations of IAA. PMID- 14520575 TI - Protein phosphorylation plays a key role in sucrose-mediated transcriptional regulation of a phloem-specific proton-sucrose symporter. AB - Assimilate partitioning refers to the systemic distribution of sugars and amino acids from sites of primary assimilation (source tissue) to import-dependent tissues and organs (sinks). One of the defining questions in this area is how plants balance source productivity with sink demand. Recent results from our laboratory showed that sucrose transport activity is directly proportional to the transcription rate of the phloem-specific proton-sucrose symporter BvSUT1 in Beta vulgaris L. Moreover, symporter gene transcription is regulated by sucrose levels in the leaf. Here we show that sucrose-dependent regulation of BvSUT1 transcription is mediated, at least in part, by a protein phosphorylation relay pathway. Protein phosphatase inhibitors decreased sucrose transport activity, symporter protein and mRNA abundance, and the relative transcription rate of the symporter gene. In contrast, protein kinase inhibitors had no effect or increased sucrose transport, protein and mRNA abundance, and transcription. Furthermore, pre-treating leaves with kinase inhibitors before feeding with sucrose blocked the sucrose-dependent decrease in symporter transcription and transport activity. The latter observation provides direct evidence for a protein phosphorylation cascade operating between the sucrose-sensor and the transcriptional regulator that controls BvSUT1 expression and, ultimately, phloem loading. PMID- 14520576 TI - Heterologous expression of a fatty acid hydroxylase gene in developing seeds of Arabidopsis thaliana. AB - Expression of a cDNA encoding the castor bean ( Ricinus communis L.) oleate Delta12-hydroxylase in the developing seeds of Arabidopsis thaliana (L.) Heynh. results in the synthesis of four novel hydroxy fatty acids. These have been previously identified as ricinoleic acid (12-hydroxy-octadec- cis-9-enoic acid: 18:1-OH), densipolic acid (12-hydroxy-octadec- cis-9,15-enoic acid: 18:2-OH), lesquerolic acid (14-hydroxy-eicos- cis-11-enoic acid: 20:1-OH) and auricolic acid (14-hydroxy-eicos- cis-11,17-enoic acid: 20:2-OH). Using mutant lines of Arabidopsis that lack the activity of the FAE1 condensing enzyme or FAD3 ER Delta 15-desaturase, we have shown that these enzymes are required for the synthesis of C20 hydroxy fatty acids and polyunsaturated hydroxy fatty acids, respectively. Analysis of the seed fatty acid composition of transformed plants demonstrated a dramatic increase in oleic acid (18:1) levels and a decrease in linoleic acid (18:2) content correlating to the levels of hydroxy fatty acid present in the seed. Plants in which FAD2 (ER Delta12-desaturase) activity was absent showed a decrease in 18:1 content and a slight increase in 18:2 levels corresponding to hydroxy fatty acid content. Expression of the castor hydroxylase protein in yeast indicates that this enzyme has a low level of fatty acid Delta12-desaturase activity. Lipase catalysed 1,3-specific lipolysis of triacylglycerol from transformed plants demonstrated that ricinoleic acid is not excluded from the sn 2 position of triacylglycerol, but is the only hydroxy fatty acid present at this position. PMID- 14520578 TI - The prevalence of musculoskeletal pain in Norwegian nurses' aides. AB - OBJECTIVES: To determine the prevalence of musculoskeletal pain in Norwegian nurses' aides and how it varies with demographic factors, number of working hours per week, and service sector. METHODS: A random sample of nurses' aides, represented by the Norwegian Union of Health and Social Workers, was mailed a questionnaire in 1999. The 6,485 vocationally active respondents who were not on leave because of illness or pregnancy comprised the sample of the present study. RESULTS: The prevalence (previous 14 days) of musculoskeletal pain was 88.8% [95% confidence interval (CI) 88.0-89.6%]. The prevalence of intense musculoskeletal pain was 51.1% (CI 49.9-52.3%). The prevalence of widespread pain was 26.6% (CI 25.5-27.7%). The prevalence of pain in the extremities increased with increasing age, whereas the prevalence of headache decreased with increasing age. Neck, shoulder, elbow, high back, and hip pains were more prevalent in women than in men. The prevalence of neck pain increased with increasing working hours per week. Low back pain was most prevalent in persons working in nursing homes. Pain in the extremities was most prevalent in persons working in nursing homes and in those who were working in somatic hospital departments for adults. Headache was most prevalent in persons working in institutions for mentally handicapped people. The prevalence of musculoskeletal pain was lowest in persons working in psychiatric and paediatric departments and the community-nursing service. CONCLUSIONS: The prevalence of musculoskeletal pain in Norwegian nurses' aides is very high. Not only the lower back but also several other regions of the body are frequently affected. The prevalence rates vary with age, gender, working hours per week, and the service sector in which the aides are working. PMID- 14520577 TI - Chronic moderate hypoxia during in ovo development alters arterial reactivity in chickens. AB - We previously observed arterial sympathetic hyperinnervation and endothelial dysfunction in the chicken embryo after exposure to chronic hypoxia. We now investigate whether changes in arterial properties could also be observed at 14 15 weeks of life. Eggs of White Leghorn chicken were incubated under normoxic or moderately hypoxic (15% O2 from days 6-19 of a 21-day incubation) conditions. Experiments were performed at 14-15 weeks of life under standard conditions (Hm: males exposed to hypoxia; Hf: females exposed to hypoxia; Nm: males exposed to normoxia; Nf: females exposed to normoxia). Body weight at hatching and at 14-15 weeks was not affected by in ovo exposure to hypoxia. Mean arterial pressure and heart rate were not significantly altered by chronic in ovo hypoxia. However, isolated femoral arteries were more sensitive to electrical stimulation (frequency in Hz of half-maximal contraction, Hm: 1.62+/-0.33, Hf: 1.92+/-0.88, Nm: 2.49+/-0.49, Nf: 2.83+/-0.31) and pharmacological stimulation of peri arterial sympathetic nerves (contraction in N/m in response to tyramine: Hm: 5.27+/-0.85, Hf: 4.10+/-0.9, Nm: 2.26+/-0.67, Nf: 3.65+/-0.51, p=0.07) after in ovo hypoxia. In side branches of the femoral artery, the effect of NO synthase blockade with L-NAME on contraction (in N/m) in response to high K+ (Hm: 0.35+/ 0.91, Hf: 1.29+/-0.36, Nm: 2.88+/-0.19, Nf: 2.79+/-0.58) and on the sensitivity to acetylcholine (DeltapD2, H: 0.32+/-0.11, N: 0.62+/-0.05) was reduced after in ovo hypoxia. The present study shows that exposure to chronic moderate hypoxia during development affects the contractile and relaxing arterial responses of 14- to 15-week-old chickens. Although hypoxia did not lead to changes in blood pressure at this age, the observed effects on arterial sympathetic and endothelial function may represent early signs of future cardiovascular abnormalities. PMID- 14520579 TI - Myocarditis associated with Hashimoto's disease: a case report. AB - We report on a sudden cardiac death case involving a 40-year-old man with no known medical history. Forensic autopsy showed lymphocytic myocarditis associated with lymphocytic thyroiditis. In both the heart and the thyroid gland, the inflammatory foci often had a nodular pattern with a germinal centre. Virological and toxicological analyses were negative. Postmortem biochemistry showed a slight increase in TSH in combination with normal T3 and T4 blood levels suggesting hypothyroidism. High titres of antiperoxidase and antithyroglobulin antibodies with normal levels of TSH receptor antibodies, in addition to biological hypothyroidism and lymphocytic inflammation were consistent with the diagnosis of Hashimoto's thyroiditis. Immunohistochemical studies excluded a lymphoma and showed no evidence of viral myocarditis. In contrast to Grave's disease, Hashimoto's thyroiditis has never been reported in association with myocarditis as a cause of sudden death. We conclude that the cardiac immunological and histological pattern, similar to that found in the thyroid gland suggests an autoimmune myocarditis. PMID- 14520580 TI - Surgical correction of deformities of the distal radius due to fractures in pediatric patients. AB - INTRODUCTION: The indication and treatment of malunited fractures of the distal radius in the growing skeleton differ from those for adults. The literature results are scarce. In this study we examined the results of surgical correction following fractures of the distal radius in infants. MATERIALS AND METHODS: Seven infant patients with malaligned fractures of the distal radius that demonstrated significant functional deficit and poor prognosis for spontaneous recovery were treated with surgical correction. Two of them were treated because of growth disturbance from post-traumatic closure of the distal radial physis. RESULTS: All seven osteotomies healed with acceptable radiologic alignment. After an average follow-up period of 3 years and 1 month (range 10 months to 8 years and 4 months), patients had an average range of wrist motion (ROM) of 131 deg in extension and flexion (93% of the contralateral side, 130% of presurgical situation); 65 deg in ulnar and radial deviation (94% of the contralateral side, 122% of presurgical situation); and 177 deg in pronation and supination of the forearm (97% of the contralateral side, 115% of presurgical situation). The average grip strength of 71.4 kPa was nearly equal to the other side with 72.8 kPa. The average postoperative angulation of the distal radius was 12 (range 10 17) deg, the radial inclination 20 (range 12-30) deg, and the ulnar variance was 0.6 (range -3 to +2) mm. According to the Fernandez point score system as well as the scale of Fernandez, there were five excellent results and two good results. The patients described low disability on the DASH scores, with a median of 4 (range 0-41) points. CONCLUSIONS: Surgical correction for malunited fractures of the distal radius provides good and excellent radiological and functional results in the growing skeleton. It should be considered immediately if there is poor remodeling capacity and disabling loss of function. PMID- 14520581 TI - Complications in high tibial (medial opening wedge) osteotomy. AB - INTRODUCTION: The high tibial (medial opening wedge) osteotomy (HTO) is a standard procedure in the treatment of varus gonarthrosis. This is potentially associated with various complications. The aim of this study was an analysis of complications and potential technical mistakes. MATERIALS AND METHODS: A total of 85 patients (49 male and 36 female) suffering from varus gonarthrosis underwent a medial opening wedge HTO. The osteotomy was fixed in 55 patients by a spacer plate (Puddu plate; group A). In group B (n=30), the osteotomies were fixed by C plate. RESULTS: The rate of complications was 43.6% in group A and 16.7% in group B (p<0.05). Infraction of the lateral tibial head is a possible intraoperative complication. This was seen in 11.7%. An additional osteosynthesis was required in group A. In contrast, the C-plate can solve this problem without additional measures. General complications of the HTO were seen: infection (4.7%), hematoma (4.7%), and thrombosis (2.3%). In every case of a severe deep infection, the osteotomy space was filled with synthetic bone graft. These grafts were used only in group A. Failure of the implants is a potential cause of loss of correction. This complication was seen nine times in group A but never in group B. CONCLUSION: A diligent surgical technique and a convenient implant are obligatory in (medial opening wedge) HTO. PMID- 14520584 TI - Transient tachypnea of the newborn (TTN): a role for polymorphisms of surfactant protein B (SP-B) encoding gene? AB - BACKGROUND: Transient tachypnea of the newborn (TTN) is usually a benign self limiting respiratory disorder in the immediate neonatal period. The lipophilic surfactant-associated protein B (SP-B) was demonstrated to be the most relevant structural component of the surfactant system for immediate postnatal pulmonary adaptation. We hypothesized genetic variations of surfactant protein B (heterozygous 121 ins 2 mutation er intron 4 polymorphisms) to be related to TTN. PATIENTS AND METHOD: We screened genomic DNA of 83 healthy term neonates (gestational age: 39 (37 - 41) completed weeks [median and range]; birth weight: 3325 +/- 541 grams [mean +/- SD]) and 75 infants presenting with TTN (gestational age: 38 (37 - 41) completed wecks [median and range]; birth weight: 3091 +/- 435 grams [mean +/- SD]) by means of PCR-amplification, fragment length and sequence analysis. TTN was diagnosed an the basis of the clinical signs with respiratory rate > 60 breaths/minute, fraction of inspired oxygen > 0.21, and characteristic radiographic findings within less than 24 hours after birth. Newborns with any infection, pulmonary or cardiac congenital malformations, postnatal asphyxia and infants born to diabetic mothers were excluded. RESULTS: In TTN-group the frequency of male infants (68.4 % versus 44.6 %, p < 0.05) and caeserian section were significantly higher (68.4 % versus 30.1 %, p < 0.05). We did not find any statistical difference in frequency of intron 4 variations between controls and TTN-group (8.4 % versus 10.7 %). None of the infants were heterozygous for the 121ins2 SP-B mutation. CONCLUSIONS: WC conclude polymorphisms of intron 4 and heterozygous 121 ins 2 mutation not to associated with TTN. PMID- 14520586 TI - [Inhalation therapy with nitric oxide in pulmonary hypertension: Comparison of preterm infants versus newborn infants]. AB - BACKGROUND: Inhaled nitric oxide (iNO) is used as a vasodilator in pulmonary hypertension (PH) of the newborn infant. PATIENTS AND METHODS: Retrospective analysis of patients, who were treated at our department with iNO in the period from 1994-2001. Response was defined as an increase of the paO (2)/FiO (2) Ratio > or = 20 % and/or a decrease of the oxygenation index (OI) >/= 20 % after 2 h (early response), and consecutively after 24 h (late response). The patients were divided into a) primary persistent pulmonary hypertension of the newbom (PPHN), or b) pulmonary hypertension secondary to meconium aspiration syndrome (MAS), sepsis or congenital diaphragmatic hernia (CDH). RESULTS: Between 1994 and 2001 we treated 47 patients with iNO at our neonatal intensive care unit. We included 16 (35 %) preterm infants (GA 34,5 [25 - 37] weeks, GG 2061 [680 - 3410] g) (Median/Range) and 31 (65 %) newbom (GA 40 [38 - 42] weeks, GG 3510 [2550 - 4560] g). 18 (38 %) patients suffered from primary PPHN, 29 (62 %) from secondary PPHN (14 MAS [30 %], 8 sepsis [17 %], 4 CDH [8 %]). 8 (50 %) preterm and 20 (64 %) term infants showed a positive iNO response after 2 h, again 8 (50 %) preterm and 20 (64%) term infants showed a positive iNO response after 24 h. There was neither a significant difference between term and preterm infants at 2 h, nor at 24 h. Between 2 h and 24 h 10 patients changed in their response to iNO. 5 (18 %) patients with early response showed a significant degradation after 24 h, whereas 5 (26 %) of the patients without early response showed a significant improvement of the oxygenation alter 24 h. Alltogether 13 (72 %) patients with PPHN, 8 (57 %) with MAS, 2 (50 %) with CDH, 4 (50 %) with sepsis showed a positive iNO response after 24 h. In regard to the oxygenation parameters at start of iNO-therapy, the patients with early response did not differ from the patient without response (median OI: 20,0 versus 21,8, median paO (2)/FiO (2) Ratio: 59,3 versus 55,0 mmHg at the start of the iNO therapy). CONCLUSION: In regard to iNO response, there was no significant difference between term and preterm infants. Due to the changing response, a positive iNO-response after 2 h had no predictive value for the further prognosis of the oxygenation situation under iNO therapy. PMID- 14520585 TI - [Transient periventricular echodensities (TPE) in preterm infants under 1500 gramms: an analysis of the outcome of the last 10 years]. AB - BACKGROUND: The aim of our study was to analyze neurodevelopmental outcome in preterm infants with TPE. PATIENTS: 46 preterm infants with a birthweight (BW) < or = 1500 g were included in the study. They all had TPE in parieto-occipital location (some combined with frontal flares). Exclusion criteria were PVL II-IV, intraventricular hemorrhage, asphyxia and isolated frontal TPE. METHOD: We analyzed ultrasound scans (1992 - 2001) of patients with TPE (diameter > 1 cm, visible in both sagittal and coronal planes, duration > 7 days), and neurological follow-up visits (date of demission, corrected age of 4, 8, 12 months, and after that every 2 years until 6 years of age) regarding the neurodevelopmental outcome. We devided the patients in group 1 (duration of TPE 8 - 14 days) and group 2 (duration of TPE > 14 days) and then analyzed the groups regarding neurodevelopmental disorders and factors increasing the risk of occurrence of TPE. RESULTS: From 1992 - 2001 578 children < or = 1500 g were admitted at our department. 48 patients (8,3 %) fulfilled the entry criteria, 2 of them died, so 46 were included in the study. In group 1 (10 patients) the median gestational age (GA) was 28 weeks (range 25 - 31), median BW 1075 g (range 685 - 1430), 6 children were neurologically unimpaired, one developed a motor handicap, and 3 a psychomotor retardation. In group 2 (36 patients) the median GA was 28 weeks (range 25 - 35), median BW 1034,5 g (range 540 - 1470), 24 patients were neurologically unimpaired, 7 developed a motor handicap, and 5 a psychomotor retardation. No statistical difference was found between the groups neither in neurodevelopmental outcome nor factors increasing the risk of the occurrence of TPE. CONCLUSIONS: Obviously, the finding of TPE is of clinical relevance already at a duration of only 7 days regarding later neurological development. PMID- 14520587 TI - [Difficulties in diagnosing Marfan's syndrome in childhood and adolescence]. AB - BACKGROUND: The diagnosis of Marfan's syndrome in childhood and adolescence is made by the criteria of the Gent nosology, which evaluates genetic data, family history and a spectrum of clinical criteria. Due to the age dependent manifestations of the clinical symptoms, combined with the extreme heterogeneity of Marfan's syndrome diagnosis in early childhood remains sometimes difficult. PATIENTS: Prospectively, we analyzed the clinical symptoms of all patients where Marfan's syndrome was suspected. We evaluated those patients between January 1997 and April 2002 by an interdisciplinary approach. METHODS: We compared the clinical datas of the patients by using the Gent nosology and the Berlin nosolgy. RESULTS: 34 patients underwent full follow-up. The median age was 10,32 years with a range of 0,01 to 37,31 years, 19 patients were male, 15 patients were female. In eight patients Marfan's syndrome could be rouled out, 9 of 26 patients (34,6 %) fullified the criteria of the Gent nosology, in 17 of 26 patients (65,4 %) Marfan's syndrome remained just suspected, but was not fullified by the criteria of the Gent nosology. Concerning the Berlin nosology 14 of 26 patients (53,8 %) fullified the criteria, 12 of 26 patients (46,2 %) failed. Due to the criteria of the Gent nosology 14 patients (53,8 %) fullified the criteria of skeletal involvement, 21 patients (80,8 %) fullified cardiovascular major manifestation, 6 patients (23,1 %) had an ophthalmic major manifestation, and 9 patients (34,6 %) had an affected first degree relative or were genetically determined. CONCLUSIONS: On the basis of the data of our patients the diagnosis of Marfan's syndrome in childhood and adolescence can be made more sensitive by the criteria of the Berlin nosology compared to the Gent nosology. This seems to be caused by the age dependent manifestations of the symptoms. Until diagnostic algorhythms of Marfan's syndrome in childhood remain suboptimal, continuous clinical follow-up for all cases even those only in the case of suspected Marfan's syndrome are necessary to exclude complicated course and to improve outcome. PMID- 14520588 TI - [Subacute sclerosing panencephalitis (SSPE)]. AB - Subacute sclerosing panencephalitis is a rare progressive neurological disorder of childhood and early adolescence caused by persistent measles virus. The diagnosis is based upon characteristic clinical manifestations, periodic EEG discharges, raised antibody titre against measles/SSPE in the plasma and cerebrospinal fluid and increase of gamma-globulins in the cerebrospinal fluid. Histopathological examination shows neuronal loss, astrogliosis, demyelination, infiltration of inflammatory cells, and intranuclear inclusions in neurons, oligodendrocytes and astrocytes. In most cases nucleocapsids are detected by electron microscopy. Although treatment is still undetermined, combination of intrathecal high-dose interferon-alpha and intravenous ribavirin administered at an early stage of SSPE seems to be effective. PMID- 14520589 TI - [Parvovirus B 19 associated hemophagocytic syndrome in a patient with hereditary sperocytosis]. AB - Infections with parvovirus B 19 can cause aplastic crises with a rapid decline of hemoglobin levels in patients with hereditary spherocytosis. Usually, the symptoms and signs of the actual infection are mild. We here report on an eight year old girl with hereditary spherocytosis who was admitted to hospital with high temperature, headache, impaired consciousness and a profound anemia (Hb 2.9 mmol/l). Since she also developed low leukocyte and platelet counts a hematological malignancy was suspected. The bone marrow aspirate showed only 1 % erythroblasts and macrophages with active hemophagocytosis. The serum ferritin was 1381,4 ng/ml. Both, serology and PCR revealed an active infection with parvovirus B 19. Coagulation analysis suggested a low degree of disseminated intravasal coagulation (low fibrinogen, high D-dimers). We diagnosed a parvovirus B 19 associated hemophagocytic syndrome. With only symptomatic treatment the patient's condition and laboratory findings improved during the course of a few days. In accordance with other reported cases, the prognosis of parvovirus B 19 associated hemophagocytic syndrome seems to be better than in hemophagocytic syndrome of other origin. PMID- 14520590 TI - [Cow milk-specific humoral and cellular immune response in infants with high risk of atopy under feeding a whey hydrolysate infant formula]. AB - BACKGROUND: Hypoallergenic infant formulas (HAF) were developed for atopy prevention in infants with high risk of atopy if these cannot be breastfed. HAF mount an antigen-specific immune response in infants. The aim of the study was to analyse the immune response in infants fed with a new infant formula based on a whey hydrolysate (HAF) and to compare it with that of exclusively breastfed controls. PATIENTS AND METHODS: Plasma concentrations of cow milk-specific IgE were analysed in 94 infants with high risk of atopy, 44 were exclusively breastfed, 50 were fed with HAF. In addition, cow milk-specific IgG antibodies (26 breastfed, 30 fed with HAF) as well as proliferation of periph-eral blood mononuclear cells to bovine beta-lactoglobulin (BLG) (41 breastfed, 47 fed with HAF) were tested. Specific IgE and IgG antibodies were determined using enzymoimmunometric assay (Alastat). Cellular proliferation was measured using tritiated thymidine incorporation assay after 6 day stimulation with BLG. RESULTS: Elevated IgE to cow milk antibodies (> 0.35 kU/L) were detected in two infants from the breastfed group and in one from the HAF-fed group. The plasma concentrations of milk specific IgG antibodies in HAF-fed infants were insignificantly higher than those in breastfed ones. No significant difference was found in bovine BLG-specific cell proliferation between both groups. CONCLUSION: Concerning the properties investigated like antigenicity, allergenicity and immunogenicity, the extensively hydrolysed whey based hypoallergenic formula does not significantly differ from mother milk in 6 month old infants with an increased atopy risk. PMID- 14520591 TI - [Papules on the dorsal interphalangeal joints and an unspecific immunostimulating agent]. AB - We report the case of an 8-year-old girl with skin eruptions on both hands that were thought to be of viral origin and, therefore, had been treated with an unspecific immunostimulating agent, containing large amounts of inosine. Under this therapy, which is - despite contrasting knowledge - still thought to be harmless and without serious side effects in the opinion of many physicians and especially medical laymen, the girl's condition worsened rapidly. Diagnosis of juvenile dermatomyositis was made. Because inosine is able to enhance T-cell proliferation and reverse immunosuppression in vitro, both mechanisms may have aggravated the disease course in our patient, once the autoimmune process of dermatomyositis has been initiated. PMID- 14520592 TI - Hypercalciuria in osteogenesis imperfecta type I. AB - BACKGROUND: In Osteogenesis Imperfecta severity of disease and reduced physical activity have been considered the main factors contributing to hypercalciuria; however, its pathogenesis in Osteogenesis Imperfecta Type I, in which mobility is normal, is still unclear. PATIENT, METHODS AND RESULTS: We describe a patient with Osteogenesis Imperfecta Type I and hypercalciuria, in whom measurement of calcium intake, plasma 1 - 25(OH) (2) Vitamin D, fasting calciuria and tubular proteinuria led us to exclude an absorptive or renal component in the pathogenesis of hypercalciuria. CONCLUSIONS: We believe that hypercalciuria is determined by bone disease in Osteogenesis Imperfecta Type I. This condition should be added to the causes of normocalcemic hypercalciuria in children and the mildest forms should be differentiated from Idiopathic Hypercalciuria. PMID- 14520593 TI - [Polio-like myelitis due to Coxsackie-Virus B 3: Course under treatment with pleconaril]. AB - Enteroviruses are common with infections of the CNS, such as encephalitis and myelitis, but they may cause various diseases in different organ systems, particulary with fatal outcome. Pleconaril is a new orally acting antiviral drug with broad anti-picornavirus activity, which provides to treat rhinoviral and enteroviral infections. To explain the importance for clinical use, we report a case of severe enteroviral infections of the CNS, treated by Pleconaril. A 14 year old girl presented with a severe polio- like myelitis including flaccid paraparesis and urinary incontinence due to Coxsackievirus-infection. Because of prolonged course and virus persistence we treated with Pleconaril, after treatment a remarkable improvement could be noticed, continency and the ability to walk without aid were regained within a few weeks. With the development of new antiviral substances we are now given more opportunities to treat infectious conditions of the central nervous system. We suggest to include enteroviruses in diagnostic procedures since there is an effective treatment with the new drug Pleconaril available. PMID- 14520594 TI - Therapy of hyperthyroidism. AB - Hyperthyroidism is a common disorder and affects approximately 2 % of women and 0.2 % of men. The review focuses on the therapy of overt hyperthyroidism with special emphasis on treatment strategies in Germany and Europe. Current treatment schedules for the different causes of hyperthyroidism are described and new therapeutic aspects are discussed. Special sections deal with the treatment of hyperthyroidism in pregnancy, neonates and children, and the treatment of thyrotoxic storm. PMID- 14520595 TI - Why is a new trial required to define the role of insulin reserve in newly diagnosed adult type 1 diabetes? PMID- 14520596 TI - Influence of etofibrate on LDL-subtype distribution in patients with diabetic dyslipoproteinemia. AB - Low density lipoprotein (LDL-) particles can be subfractionated in large-buoyant (lb), intermediate-dense (id) and small-dense (sd) LDL-subtypes. Fibrates improve the LDL-subtype profile by reducing proatherogenic sd-LDL which are prominent in diabetic dyslipoproteinemia. We evaluated the effect of etofibrate on the LDL subtype distribution in patients with type 2 diabetes mellitus (n = 13, 55 +/- 18 years, BMI 27.9 +/- 5.5 kg/m2, HbA1c 10.1 +/- 3.9 %) and diabetic dyslipoproteinemia (triglycerides 343 +/- 253 mg/dl, HDL-cholesterol 36 +/- 7 mg/dl, LDL-cholesterol 110 +/- 37 mg/dl). Plasma lipids (enzymatic methods) and LDL-subtypes (7 LDL-subfractions, density gradient ultracentrifugation) were measured before and during etofibrate therapy (500 mg/d, 7 - 16 weeks). Etofibrate significantly (p < 0.05, Wilcoxon-test) reduced triglycerides (- 31 +/ 60 %) and increased HDL-cholesterol (+ 24 +/- 22 %), whereas total cholesterol and LDL-cholesterol did not change. Cholesterol concentration decreased in sd-LDL by 12 % (p < 0.05), while it increased in id- and lb-LDL (+ 26 %,+ 39 %, respectively). Thus, the LDL-subtype profile showed a relative increase of the fraction of lb- (+ 13 +/- 32 %, n.s.) and id-LDL (+ 23 +/- 33 %, p < 0.05) and a relative decrease of the fraction of sd-LDL (- 19 +/- 18 %, p < 0.05). We conclude that etofibrate not only decreases triglycerides and increases HDL cholesterol but also improves the LDL-subtype profile and thus may reduce the cardiovascular risk in patients with an abundance of sd-LDL such as diabetic patients. PMID- 14520597 TI - Influence of iodide and iodolactones on thyroid apoptosis. Evidence that apoptosis induced by iodide is mediated by iodolactones in intact porcine thyroid follicles. AB - Iodine induced thyroid involution is caused by apoptosis rather than necrosis. This effect of iodide on apoptosis of thyroid epithelial cells may be not a direct one but mediated by iodinated derivatives i.e. of polyunsaturated fatty acids, especially of iodolactones, which have previously shown to inhibit thyroid cell proliferation. We studied the influence on apoptosis of iodide (2 microM and 20 microM) and iodolactone (0.05 microM and 0.5 microM), with and without TSH (1 mU/ml), using a well characterized ex vivo- culture system of intact porcine thyroid follicles in three-dimensional culture. Apoptosis and necrosis was evaluated by electron-microscopy. Stimulation with 2 and 20 microM iodide rapidly induced a rate of apoptosis (4 - 6 %) comparable to about 40-fold lower doses of delta-iodolactone (0.05 microM and 0.5 microM). Addition of TSH (1 mU/ml) caused a slight but not significant further increase of the incidence of apoptotic cells. The rate of necrotic thyroid epithelial cells (1 - 2 %) was similar in all experiments. As delta-iodolactone in very low concentrations--comparable to iodide in higher concentrations--not only inhibits growth but also induces apoptosis, it has to be supposed that the effect of iodide is mediated by this iodinated compound. However, further experiments are necessary to confirm this hypothesis. In addition it could be demonstrated, that apoptosis is a very rapid and limited process in intact follicles. This also may explain, why iodine supplementation even in high doses does not lead to thyroid atrophy but only normalisation of thyroid size. These results confirm that apoptosis is an important regulated and limited mechanism in goiter involution. PMID- 14520598 TI - Perioperative dexamethasone treatment in childhood craniopharyngioma--influence on short-term and long-term weight gain. AB - The substitution of dexamethasone during and after surgery of childhood craniopharyngioma is necessary in order to treat and/or prevent brain edema and adrenal insufficiency. Early post-operative weight gain is a predictor for severe obesity during long-term follow-up. In a retrospective analysis of 60 patients with childhood craniopharyngioma we inquired whether dose and duration of perioperative dexamethasone therapy (n = 68) had influence on short-term post operative weight gain and long-term development of severe obesity. The median follow-up period was 4.2 years, ranging from 1 to 9 years. 24 patients (14 f/10 m) developed severe obesity (BMI > 3 SD). 28 patients (10 f/18 m) retained normal weight (BMI < 2 SD). Eight patients presented with a BMI between 2 and 3 SD at the final visit. Differences in terms of age at surgery or follow-up period were non-detectable between the analyzed groups of craniopharyngioma patients. Duration and cumulative dexamethasone doses (mg/m2 BSA) for perioperative dexamethasone therapy were similar for severely obese patients (duration: 8.7 d; 4.5 - 17 d, cumulative dose: 74; 42 - 177 mg/m2 BSA) and normal weight patients (duration: 10.0 d; 1 - 41 d; dose: 76; 9 - 390 mg/m2 BSA). Whereas cumulative dexamethasone doses positively (p < 0.01; rho: 0.424) correlated with weight gain during the first year following surgery, long-term development of severe obesity was not influenced by dose and duration of perioperative dexamethasone treatment. Patients who developed severe obesity during follow-up had a higher (p < 0.001) BMI already at the time of diagnosis. We conclude that dose and duration of perioperative dexamethasone treatment had short-term effects on post-operative weight gain, but not on the development of long-term severe obesity. The results of our retrospective analysis are currently tested in a prospective surveillance study Kraniopharyngeom 2000 (www.kraniopharyngeom.com). PMID- 14520599 TI - The role of GabaA receptors in the neural systems of the ventromedial hypothalamus-nucleus infundibular region in the control of GnRH release in ewes during follicular phase. AB - To examine the role of the GABAA receptor mediating systems in the control of gonadotropin releasing hormone (GnRH) release from the ventromedial-infundibular region (VEN/NI) of the hypothalamus of ewes during the follicular phase of the estrous cycle, the extracellular concentrations of GnRH, beta-endorphin (B-END), noradrenaline (NE), dopamine (DA), and their metabolites MHPG, DOPAC and concentration of luteinizing hormone (LH) in blood plasma were quantified during local stimulation or blockade of GABAA receptors with muscimol and bicuculline, respectively. Stimulation of GABAA receptors attenuated GnRH and LH release, increased beta-endorphin outflow and dopaminergic activity but had no evident effect on noradrenergic activity. Blockade of GABAA receptors decreased beta endorphin release but had no evident effect on the extracellular concentration of GnRH, LH levels in the blood and catecholaminergic activity. It is suggested that suppression of GnRH/LH release under muscimol treatment may result from activation of GABAA receptors on GnRH nerve terminals and through GABAA receptor mechanism activated beta-endorphinergic and dopaminergic neurons in the VEN/NI. Lack of changes in NE and MHPG concentration during stimulation or blockade of GABAA receptors suggests, that during the follicular phase of the estrous cycle the noradrenergic system in the VEN/NI is not involved in the control of GnRH/LH release by GABA. PMID- 14520600 TI - Licorice consumption and serum testosterone in healthy man. AB - We have previously found that licorice can reduce serum testosterone in healthy men. These results were not confirmed in another study, where the same amounts of licorice did not decrease salivary testosterone values. In the actual study we treated more cases with the same amount of licorice and reproduced our previous data. The mean testosterone values decreased by 26 % after one week of treatment (p < 0.01). There was also a significant increase in 17-OHP and LH concentrations and a slight, but not significant decrease in free testosterone. Licorice treatment, in addition, did not affect the response of testosterone and 17-OHP to stimulation with beta-HCG. PMID- 14520601 TI - Influence of an antidiabetic treatment with sulfonylurea drugs on long-term survival after acute myocardial infarction in patients with type 2 diabetes. The LAngendreer Myocardial infarction and Blood glucose in Diabetic patients Assessment (LAMBDA). AB - INTRODUCTION: Patients with type 2 diabetes show a significantly higher mortality after acute myocardial infarction than non-diabetic patients. The influence of sulfonylureas on the survival after acute myocardial infarction is still under debate. PATIENTS AND METHODS: Survival of 562 patients, consecutively admitted to an intensive care unit with the diagnosis acute myocardial infarction, was prospectively assessed for > 3 years. At the time of hospital admission, patients were grouped as (a) non-diabetic patients; (b) patients with newly diagnosed type 2 diabetes; (c) patients with known type 2 diabetes not treated with sulfonylureas and (d) patients with known type 2 diabetes treated with sulfonylureas. Survival-analysis was performed according to Kaplan-Meier. RESULTS: 324 patients were non-diabetics, in 86 cases type 2 diabetes was newly diagnosed at the time of hospital admission, 77 patients with known diabetes had taken sulfonylureas (glibenclamide in all cases) prior to the acute myocardial infarction, 75 patients were on any other antidiabetic treatment. Long-term survival was significantly shorter in patients with type 2 diabetes compared to the non-diabetic patients (p < 0.0001). However, no significant differences were observed between the patients with type 2 diabetes treated with sulfonylurea drugs and those receiving any other antidiabetic treatment (p = 0.53) CONCLUSIONS: An antidiabetic treatment with sulfonylurea-drugs prior to acute myocardial infarction does not have negative effects on the long-term survival. Larger prospective studies will be necessary to finally clarify this question. PMID- 14520602 TI - The connection between QT dispersion and autonomic neuropathy in patients with type 2 diabetes. AB - It is controversial whether an increase in the QT dispersion (QTd) on the electrocardiogram (ECG) reflects cardiac autonomic neuropathy in diabetic patients. In the current study, the QTd was compared in 60 type 2 diabetic patients and in 30 healthy subjects, and its association with autonomic neuropathy in diabetic patients was investigated. An increased QTd was found in diabetic patients, compared to healthy subjects. The QTd had significant negative associations with the log of the low and high frequency power (log LF and HF, respectively) of the power spectral analyses (PSA) of heart rate variations and the systolic blood response during standing (Delta BP). There was no significant difference in the QTd between patients with and without sympathetic skin response (SSR), reflecting peripheral sympathetic function. A significant positive correlation was also found between QTd and the systolic blood pressure (SBP). On the other hand, there was no correlation between QTd and serum total cholesterol (TC), triglycerides (TG), fasting plasma glucose (FPG), hemoglobin (Hb) A (1C) concentrations or body mass index (BMI). By multiple regression analysis, the log HF, which reflects cardiac parasympathetic function, and the SBP alone were significantly associated with QTd as the dependent variable. The Delta BP and log LF, which partially reflect sympathetic nerve function, had no significant associations with QTd. These findings suggest that QTd reflects cardiac autonomic neuropathy (relative parasympathetic neuropathy) and that the QTd is also influenced by SBP, independent of autonomic neuro-function. PMID- 14520603 TI - Direct comparison of inositol phosphoglycan with prostaglandylinositol cyclic phosphate, two potential mediators of insulin action. AB - Though insulin signalling is thought by many groups to function without second messenger action, others have provided evidence for the existence and action of such regulators. Chemically quite different compounds, however, have been proposed as mediators, such as various inositol phosphoglycans and prostaglandylinositol cyclic phosphate (cyclic PIP). In spite of marked structural differences, these compounds are reported to have the same regulatory properties, i.e. to activate protein ser/thr phosphatases and to inhibit protein kinase A. In order to clarify this discrepancy, the regulatory potency of these different compounds was assayed under identical conditions. It was found that in contrast to cyclic PIP, the synthetic inositol phosphoglycan PIG41 neither directly inhibited protein kinase A nor activated protein ser/thr phosphatases. However, when added to intact cells, such as primary adipocytes, PIG41 inhibited isoproterenol-stimulated lipolysis. This effect most likely results from tyrosine phosphorylation of insulin receptor substrates (IRSs) by PIG41. This tyrosine phosphorylation is not carried out by the insulin receptor tyrosine kinase but by cytosolic tyrosine kinases. This indicates that cyclic PIP, an intracellular regulator, which primarily acts on protein kinase A and on protein ser/thr phosphatases, operates more downstream in the signal transduction cascade as compared to the inositol phosphoglycan PIG41. Thus, cyclic PIP appears to be a suitable candidate to close the gap between IRSs and the protein kinases/phosphatases involved in the signal transduction of insulin. PMID- 14520604 TI - Clinical characterisation of severe hypoglycaemia--a prospective population-based study. AB - AIM: To determine the clinical characteristics of severe hypoglycaemia (SH) in a nonselected German population. SH was defined as an event requiring intravenous glucose or glucagon injection. METHODS: The prospective population-based study screened sensitively for SH in a region with 200,000 inhabitants between 1997 and 2000. All 30,768 patients who presented to the regional central hospital emergency department, and 6,631 (85 %) of 7,804 patients attended by the emergency medical service in the region were given an initial blood glucose test to detect atypical hypoglycaemia. RESULTS: Altogether, 264 cases of SH were registered, which occurred either spontaneously (n = 14; 5 %), in subjects with type 1 (n = 92; 35 %) or type 2 diabetes (n = 148; 56 %), or in subjects with a non-classified form of diabetes (n = 10; 4 %). On the basis of the estimated local number of diabetic patients the annual rate of SH was 1.5 episodes per 100 patients in insulin-treated type 2 diabetics compared with a rate of 0.4 episodes per 100 patients for the overall group of type 2 diabetic patients. Nocturnal hypoglycaemia accounted for 44 % of episodes in patients with type 1 diabetes on intensified therapy but for only 25 % in patients with type 2 diabetes. 26 % of the hypoglycaemic individuals with type 1 diabetes had an impaired awareness of hypoglycaemia and thus recurrent hypoglycaemic episodes. Irrespective of the treatment, the most frequent contributing factors for SH in type 2 diabetic patients were advanced age (76 +/- 12 years), multimorbidity (3.6 +/- 2.6 concomitant diseases)--in particular renal impairment (54 % [80/148])--and polypharmacy (4 +/- 2.7 concomitant drugs). 34 % (50/148) of the subjects with type 2 diabetes lived in nursing homes or were cared for by a home nursing service. With standardised treatment zero mortality of SH in diabetic patients was achieved, only one non-diabetic died due to hepatic failure. CONCLUSION: In elderly, multimorbid patients approaching the insulin-deficient end of the spectrum of type 2 diabetes the risk of developing SH increases considerably, nearing that in patients with type 1 diabetes. In order to avoid SH in geriatric patients, the treatment targets should be defined critically, taking into account individual quality of life and life expectancy. Hypoglycaemia unawareness is a major risk factor for SH in type 1 diabetes. PMID- 14520605 TI - Lack of effect of acute repaglinide administration on postprandial lipaemia in patients with type 2 diabetes mellitus. AB - The effect of acute repaglinide administration (2 mg) on postprandial glycaemia and lipaemia has been examined in 20 subjects with type 2 diabetes mellitus. Each subject received in the morning, after a 12 to 14 h fast, a standard mixed meal (total energy 783 kcal), preceded by one tablet of 2 mg repaglinide or placebo. Chylomicrons and chylomicron-deficient plasma were prepared by ultracentrifugation. Triglyceride levels in CM fraction (CM-triglycerides) in total plasma as well as in CM-deficient plasma (non-CM-triglycerides) were determined. A significant reduction in postprandial glycaemia was observed after repaglinide administration compared to placebo ( p < 0.001). Plasma concentrations of total triglycerides, CM-triglycerides, non-CM-triglycerides, free fatty acids and the other plasma lipids measured, were not significantly different between the two phases of the study. It is concluded that, in contrast to sulphonylureas, acute repaglinide administration does not improve postprandial lipaemia in patients with type 2 diabetes. PMID- 14520606 TI - Graves' disease coexisting with probable autoimmune hypoparathyroidism. AB - Organ-specific autoimmune endocrine disorders may present as single diseases or may occur together in polyendocrine syndromes. We present a report of 23-year-old female with Graves' disease and concurrent hypocalcemia. As she lacked other specific features of autoimmune polyendocrine syndromes, the most likely diagnosis was Graves' disease coexisting with autoimmune hypoparathyroidism. PMID- 14520607 TI - K/DOQI clinical practice guidelines for bone metabolism and disease in chronic kidney disease. PMID- 14520612 TI - Proteinuria and other markers of chronic kidney disease: a position statement of the national kidney foundation (NKF) and the national institute of diabetes and digestive and kidney diseases (NIDDK). PMID- 14520613 TI - Should the K/DOQI definition of chronic kidney disease be changed? PMID- 14520614 TI - Should the K/DOQI definition of chronic kidney disease be changed? PMID- 14520615 TI - Hepatitis C and renal disease: an update. AB - Hepatitis C is both a cause and a complication of chronic renal disease. Chronic infection with hepatitis C virus (HCV) can lead to the immune complex syndromes of cryoglobulinemia and membranoproliferative glomerulonephritis (MPGN). The pathogenetic mechanisms for these conditions have not been defined, although they are clearly caused by the chronic viral infection. Management of HCV-related cryoglobulinemia and MPGN is difficult; antiviral therapy is effective in clearing HCV infection in a proportion of patients, but these conditions can be severe and resistant to antiviral therapy. Hepatitis C also is a complicating factor among patients with end-stage renal disease and renal transplants. The source of HCV infection in these patients can be nosocomial. Screening and careful attention to infection control precautions are mandatory for dialysis units to prevent the spread of hepatitis C. Prevention of spread is particularly important in these patients because HCV infection is associated with significant worsening of survival on dialysis therapy, as well as after kidney transplantation. Furthermore, therapy for hepatitis C is problematic, only partially effective, and associated with significant side effects in this population. There are significant needs in both basic and clinical research in the pathogenesis, natural history, prevention, and therapy for hepatitis C in patients with renal disease. PMID- 14520616 TI - Tamm-Horsfall glycoprotein: biology and clinical relevance. AB - Tamm-Horsfall glycoprotein (THP) is the most abundant urinary protein in mammals. Urinary excretion occurs by proteolytic cleavage of the large ectodomain of the glycosyl phosphatidylinositol-anchored counterpart exposed at the luminal cell surface of the thick ascending limb of Henle's loop. We describe the physical chemical structure of human THP and its biosynthesis and interaction with other proteins and leukocytes. The clinical relevance of THP reported here includes: (1) involvement in the pathogenesis of cast nephropathy, urolithiasis, and tubulointerstitial nephritis; (2) abnormalities in urinary excretion in renal diseases; and (3) the recent finding that familial juvenile hyperuricemic nephropathy and autosomal dominant medullary cystic kidney disease 2 arise from mutations of the THP gene. We critically examine the literature on the physiological role and mechanism(s) that promote urinary excretion of THP. Some lines of research deal with the in vitro immunoregulatory activity of THP, termed uromodulin when isolated from urine of pregnant women. However, an immunoregulatory function in vivo has not yet been established. In the most recent literature, there is renewed interest in the capacity of urinary THP to compete efficiently with urothelial cell receptors, such as uroplakins, in adhering to type 1 fimbriated Escherichia coli. This property supports the notion that abundant THP excretion in urine is promoted in the host by selective pressure to obtain an efficient defense against urinary tract infections caused by uropathogenic bacteria. PMID- 14520617 TI - A population-based study of the incidence and outcomes of diagnosed chronic kidney disease. AB - BACKGROUND: This study aims to determine the incidence rate and prognosis of detected chronic kidney disease (CKD) in a defined population. METHODS: This is a retrospective cohort study of all new cases of CKD from Southampton and South West Hampshire Health Authority (population base, 405,000) determined by a persistently increased serum creatinine (SCr) level (>or=1.7 mg/dL [>or=150 micromol/L] for 6 months) identified from chemical pathology records. Follow-up was for a mean of 5.5 years for survival, cause of death, and acceptance to renal replacement therapy (RRT). RESULTS: The annual incidence rate of detected CKD was 1,701 per million population (pmp; 95% confidence interval [CI], 1,613 to 1,793) and 1,071 pmp (95% CI, 1,001 to 1,147) in those younger than 80 years. There was a steep age gradient; median age was 77 years. The man-woman rate ratio was 1.6 (95% CI, 1.4 to 1.8), with a male excess in all age groups older than 40 years. Incidence increased in areas with greater socioeconomic deprivation. Median survival was 35 months. Age, SCr level, and deprivation index were all significantly associated with survival. Standardized mortality ratios were 36 fold in those aged 16 to 49 years, 12-fold in those aged 50 to 64 years, and more than 2-fold in those older than 65 years. Cardiovascular disease (CVD) was the most common cause of death (46%). Only 4% of patients were accepted to RRT. CONCLUSION: The incidence of diagnosed CKD is common, especially in the elderly, and is greater in more deprived areas. Prognosis is poor, with CVD prominent. More research is needed to assess the effectiveness and costs of increasing referral to nephrologists of patients with CKD. PMID- 14520618 TI - Urinary albumin to osmolality ratio predicts 24-hour urine albumin excretion in diabetes mellitus. AB - BACKGROUND: Urinary albumin to creatinine ratio (ACR) in a single urine sample has been proposed to provide an estimate of microalbuminuria by adjusting for variability in urine concentrations. We hypothesized that adjusting the urine albumin concentration of single-void specimens for actual urine osmolality (urinary albumin to osmolality ratio [AOR]) may provide a more accurate estimate of 24-hour urine albumin excretion rates (AERs). METHODS: Patients with diabetes mellitus (DM; n = 136) had urinary concentrations of albumin, glucose, and creatinine and osmolality measured on single-void samples, and albumin levels, on 24-hour samples. Microalbuminuria is defined as an AER between 30 and 300 mg/d. RESULTS: Correlation between AOR on single-void samples and AER on 24-hour samples (r = 0.87; P < 0.001) was similar to that between ACR and AER (r = 0.88; P < 0.001). Using a cutoff value of 18.4 mg/kg/mOsm x 10(2) (18.4 mg/mmol x 10(2)) for AOR resulted in a sensitivity and specificity of 82% and 86% in detecting microalbuminuria, respectively. The area under the curve (AUC) for AOR was 0.89. Using a cutoff value of 15.0 mg/g (1.7 mg/mmol) for ACR resulted in a sensitivity and specificity of 85% and 85% in detecting microalbuminuria, respectively. The AUC for ACR was 0.90. The ability of AOR to predict AER was maintained at varying degrees of glycosuria (glucose < 100 mg/dL [<5.5 mmol/L]; r = 0.77; 100 to 750 mg/dL [5.5 to 42 mmol/L]; r = 0.85; and >750 mg/dL [>42 mmol/L]; r = 0.92). CONCLUSION: Urinary AOR correlates closely with 24-hour microalbuminuria determination, and the correlation is not appreciably affected by glycosuria. Thus, AOR can be used as an alternative test to ACR in the assessment of microalbuminuria in the population with DM. PMID- 14520619 TI - Predialysis psychoeducational intervention and coping styles influence time to dialysis in chronic kidney disease. AB - BACKGROUND: Consensus endorses predialysis intervention before the onset of end stage renal disease. In a previous study, predialysis psychoeducational intervention (PPI) extended time to dialysis therapy by a median of 6 months. We undertook to replicate and extend this finding by examining hypothesized mechanisms. METHODS: We used an inception-cohort, prospective, randomized, controlled trial with follow-up to evaluate an intervention that included an interactive 1-on-1 slide-supported educational session, a printed summary (booklet), and supportive telephone calls once every 3 weeks. Participants were sampled from 15 Canadian (tertiary care) nephrology units and included 297 patients with progressive chronic kidney disease (CKD) expected to require renal replacement therapy (RRT) within 6 to 18 months. The main outcome was time to dialysis therapy (censored at 18 months if still awaiting RRT). RESULTS: Time to dialysis therapy was significantly longer (median, 17.0 months) for the PPI group than the usual-care control group (median, 14.2 months; Cox's proportional hazards analysis, controlling for general nonrenal health, P < 0.001). Coping by avoidance of threat-related information (called blunting) was associated with shorter times to dialysis therapy (P < 0.032). A group x blunting interaction (P < 0.069) indicated: (1) time to dialysis therapy was shortened in the usual-care group, especially when patients coped by blunting; but (2) time to dialysis therapy was extended with PPI, even among patients who coped by blunting. Knowledge acquisition predicted time to dialysis therapy (r = 0.14; P < 0.013). Time to dialysis therapy was unrelated to depression or social support. CONCLUSION: PPI extends time to dialysis therapy in patients with progressive CKD. The mechanism may involve the acquisition and implementation of illness related knowledge. Routine follow-up also may be especially important when patients cope by avoiding threat-related information. PMID- 14520620 TI - Circulating endothelial cells as potential markers of the state of the endothelium in hemodialysis patients. AB - BACKGROUND: Patients with chronic kidney disease (CKD) are at high risk for atherosclerotic cardiovascular disease (ACVD). In addition to the same epidemiological risk factors of the general population, factors unique or secondary to the uremic milieu may contribute to excess cardiovascular morbidity and mortality. Recent studies suggest the endothelium has a key role in the development of atherosclerosis. Circulating endothelial cells (CECs) may be a tool to study the state of the endothelium, with their number markedly increased in conditions associated with a high degree of endothelial cell activation and/or injury. Our hypothesis is that CEC number increases in diseases that commonly progress to stage 5 CKD, as well as in patients on hemodialysis (HD) therapy, reflecting ongoing endothelial cell activation and/or injury. METHODS: The study population consisted of 22 healthy nonsmoking individuals, 29 individuals undergoing long-term HD treatment, 10 individuals with the diagnosis of diabetes mellitus and stage 1 or 2 CKD, and 7 individuals with hypertension and stage 1 or 2 CKD. The number of CECs was enumerated in all study groups. RESULTS: We found that CEC number was increased in HD patients, hypertensive patients, and patients with diabetes compared with healthy individuals. In addition, although CEC number did not accurately reflect the presence of ACVD, the number was increased significantly in a group of HD patients with active ACVD, whereas it was decreased significantly in a group of HD patients with stable ACVD. CONCLUSION: Our results suggest that CEC number may reflect ACVD activity in HD patients, independent of the presence of ACVD. PMID- 14520621 TI - ESRD patient quality of life: symptoms, spiritual beliefs, psychosocial factors, and ethnicity. AB - BACKGROUND: Recent research suggests that patients' perceptions may be more important than objective clinical assessments in determining quality of life (QOL) for patients with end-stage renal disease (ESRD). METHODS: We interviewed 165 hemodialysis patients from 3 sites using a QOL questionnaire that included the Satisfaction With Life Scale (SWLS) and the McGill QOL (MQOL) scale, which includes a single-item global measure of QOL (Single-Item QOL Scale [SIS]). The MQOL scale asks patients to report their most troublesome symptoms. We also initiated the use of a Support Network Scale and a Spiritual Beliefs Scale. RESULTS: Mean patient age was 60.9 years, 52% were men, 63% were white, and 33% were African American. Patients had a mean treatment time for ESRD of 44 months, mean hemoglobin level of 11.8 g/dL (118 g/L), mean albumin level of 3.7 g/dL (37 g/L), and mean Kt/V of 1.6. Forty-five percent of patients reported symptoms. Pain was the most common symptom (21% of patients). There was an inverse relationship between reported number of symptoms and SWLS (P < 0.01), MQOL scale score (P < 0.001), and SIS (P < 0.001). The Spiritual Beliefs Scale correlated with the MQOL scale score, SWLS (both P < 0.01), and SIS (P < 0.05). The Support Network Scale score correlated with the MQOL Existential (P = 0.01) and MQOL Support (P < 0.01) subscales. No clinical parameter correlated with any measure of QOL, spiritual beliefs, or social support. CONCLUSION: Symptoms, especially pain, along with psychosocial and spiritual factors, are important determinants of QOL of patients with ESRD. Additional studies, particularly a longitudinal trial, are needed to determine the reproducibility and utility of these QOL measures in assessing patient long-term outcome and their association with other QOL indices in larger and more diverse patient populations. PMID- 14520622 TI - Salivary and lacrimal secretion is reduced in patients with ESRD. AB - BACKGROUND: A reduction in salivary and lacrimal secretion has been described in several diseases. However, such alterations have not been investigated fully in patients with chronic renal failure. The aim of the present study is to estimate the frequency of alterations in salivary and lacrimal secretion in long-term hemodialysis patients. METHODS: Sixty-three hemodialysis patients and 23 healthy control subjects were studied. In all of them, we tested salivary secretion (Saxon's test), lacrimal secretion (Shirmer's test), and the presence of xerostomia and xerophthalmia symptoms. In a subgroup of patients, we performed other tests to evaluate evidence of ocular lesions and tissue damage to salivary glands. We also tested the relationship between salivary and lacrimal secretion and autonomic nervous system function. RESULTS: On average, salivary and lacrimal secretion were markedly reduced in uremic patients compared with healthy controls, and alterations in salivary gland function were related strongly to salivary gland fibrosis and atrophy. Xerophthalmia often was asymptomatic, but frequently was associated with corneal lesions. Xerostomia and xerophthalmia were unrelated to autonomic dysfunction and hepatitis C virus infection. CONCLUSION: A reduction in lacrimal and salivary secretion is frequent in long-term dialysis patients. Such alterations often are asymptomatic and could be the expression of acceleration of an age-dependent decline in glandular function and attendant fibrosis and atrophy. PMID- 14520623 TI - Changes of bone remodeling immediately after parathyroidectomy for secondary hyperparathyroidism. AB - BACKGROUND: Successful parathyroidectomy for secondary hyperparathyroidism alleviates bone pain and is followed by the development of hypophosphatemia and hypocalcemia, as well as an increase in bone mineral density. An increase in osteoblast surface (Ob.S/BS) is not observed several months after surgery. In this study, we investigated early bone changes at 1 week after parathyroidectomy and the mechanism underlying an increase in bone mineral density. METHODS: Fourteen patients with severe secondary hyperparathyroidism underwent iliac bone biopsy before and 1 week after parathyroidectomy. Changes in histomorphometric parameters, including osteoclast surface (Oc.S/BS), eroded surface (ES/BS), erosion depth (E.De), fibrosis volume (Fb.V/TV), Ob.S/BS, osteoid volume (OV/BV), osteoid surface (OS/BS), and osteoid thickness (O.Th), were investigated. Changes in texture of mineralized bone and osteoid seams were also investigated. RESULTS: Oc.S/BS (P < 0.001), ES/BS (P < 0.01), and E.De (P < 0.001) decreased, but Fb.V/TV did not change at 1 week postoperatively. In particular, osteoclasts disappeared in almost all patients. Ob.S/BS (P < 0.001) increased, and cuboidal osteoblasts were proliferating on the trabecular surface where osteoclasts had existed before parathyroidectomy. As a result, newly developed osteoblasts coexisted with fibrous tissue after surgery. OV/BV (P < 0.005), OS/BS (P < 0.005), and O.Th (P < 0.005) increased, with lamellar osteoid volume showing a particular increase. Bone mineralization continued despite the low postoperative serum parathyroid hormone level. CONCLUSION: A rapid decrease in serum parathyroid hormone level after parathyroidectomy appears to suppress bone resorption, as well as cause a transient marked increase in bone formation and an increase in normal lamellar osteoid seams. PMID- 14520624 TI - Clinical experience with percutaneous ethanol injection therapy in hemodialysis patients with renal hyperparathyroidism. AB - BACKGROUND: Percutaneous ethanol injection therapy (PEIT) is a noteworthy method to treat patients with renal hyperparathyroidism (RHPT). This study was performed to enable the authors to propose appropriate indications for PEIT. METHODS: The authors injected anhydrous ethanol under ultrasonographic guidance into the parathyroid glands of 74 patients with RHPT. The size, volume, and blood vessel density of all parathyroid glands detected were evaluated. RESULTS: The patients' mean age and hemodialysis duration were 53.8 and 14.3 years, respectively. Intact parathyroid hormone (PTH) levels decreased significantly after treatment (from 950.5 +/- 448.1 to 532.0 +/- 393.2 pg/mL [ng/L], P < 0.05), and the serum calcium and phosphorus levels as well as all bone metabolic parameters had improved. The percentage reduction was greater in cases with 1 gland over 10 mm in maximal diameter, and, as the number of glands over 10 mm rose, the percentage reduction declined. Group A was defined as patients with a small number of glands (1 or 2) with high-grade blood vessel density, and group B as patients with a larger number of such glands. The percentage reduction in intact PTH levels was significantly greater in group A than in group B (56.5 +/- 27.5% v 28.1 +/- 22.0%; P < 0.01). CONCLUSION: the authors' analysis showed that the effectiveness of PEIT was influenced by both the number of glands detected that were more than 10 mm in maximal diameter and by the grade of blood vessel density. They identified appropriate indications for PEIT based on the predictive factors that influenced the efficacy of PEIT. PMID- 14520625 TI - Utility of in vitro closure time test for evaluating platelet-related primary hemostasis in dialysis patients. AB - BACKGROUND: The platelet aggregation and skin bleeding time (SBT) tests currently used for assessment of hemostasis impairment in dialysis patients have important disadvantages. The authors explored the utility of a novel in vitro closure time test (PFA-100, platelet function analyzer) in which the process of platelet adhesion and aggregation after vascular injury is simulated in vitro in dialysis patients. METHODS: Thirty-four long-term dialysis patients were included in the study with 30 healthy volunteers as the control group. In vitro closure time was compared with results from the platelet aggregation and SBT tests. RESULTS: In vitro closure time identified more patients and fewer controls with hemostasis impairment. In the patient group, 60%, 40%, and 20%, and in the control group, 0%, 10% and 3% of persons were found to have hemostasis impairment as determined by in vitro closure time, platelet aggregation, and SBT, respectively. In addition, values for patients and controls were significantly different for in vitro closure time (P < 0.05) but not for platelet aggregation or SBT. Thus, closure time appears to be more sensitive and specific than the other 2 tests. No correlation was found between the 3 tests, either in patients or in controls. However, a high correlation (r = 0.73; P < 0.0001) was found between the 2 types of in vitro closure time test (collagen/epinephrine [CEPI] and collagen/adenosine diphosphate [CADP]) in patients and controls. CONCLUSION: These results indicate that in vitro closure time can be a useful test for detecting platelet-related primary hemostasis defects in dialysis patients. PMID- 14520626 TI - In vivo validation of glucose pump test for measurement of hemodialysis access flow. AB - BACKGROUND: The glucose pump test (GPT) is a recently introduced method of measuring hemodialysis access blood flow (Qa). A validation of GPT during dialysis has not yet been done, and performance characteristics of the method have not yet been fully analyzed. METHODS: The authors studied 33 patients (25 synthetic grafts, 8 autogenous arteriovenous fistulae). Qa measurements by ultrasound dilution (UD) and GPT were done in triplicate during dialysis. In GPT, a baseline blood sample (C(1)) was obtained, followed by infusion of a 10% glucose solution (C(i)) through the arterial needle into the access at 16 mL/min (Q(i)). After 11 seconds, a downstream blood sample (C(2)) was aspirated from the venous needle. C(1) and C(2) glucose were measured by glucometer. Qa was computed by the equation: Qa = Q(i)(C(i) - C(2))/(C(2) - C(1)). A model of the access vascular circuit was used to determine the influence of C(2) aspiration on the Qa measurement. RESULTS: Mean Qa was 1413 mL/min by UD versus 1,496 mL/min by GPT (P = 0.11). There was a strong linear correlation between the 2 methods (r = 0.905; P <0.001). The pooled coefficient of variation was 6.4% for UD and 9.6% for GPT. The circuit model showed that aspiration of C(2) causes an increase in Qa (DeltaQa) that depends on the aspiration rate (Q(ASP)) and fraction of resistance in the circuit that is downstream to the venous needle: DeltaQa = Q(ASP)(Downstream resistance)/(Total resistance). The model predicts the overestimate is approximately 62 mL/min for grafts and 120 mL/min for fistulae but may vary depending on the balance of resistances upstream and downstream to the venous needle. CONCLUSION: This study shows that GPT closely correlates with UD, and the method has adequate precision. GPT is an inexpensive method that may help make Qa measurements more widely available than previously possible. PMID- 14520627 TI - Effect of malnutrition-inflammation complex syndrome on EPO hyporesponsiveness in maintenance hemodialysis patients. AB - BACKGROUND: Elements of malnutrition-inflammation complex syndrome (MICS) may blunt the responsiveness of anemia of end-stage renal disease (ESRD) to recombinant human erythropoietin (EPO). METHODS: The authors examined cross sectional associations between the required dose of EPO within a 13-week interval as prescribed by practicing nephrologists who were blind to the study and several laboratory values known to be related to nutrition and/or inflammation, as well as the malnutrition-inflammation score (MIS), which is a fully quantitative assessment tool based on the subjective global assessment of nutrition. RESULTS: A total of 339 maintenance hemodialysis (MHD) outpatients, including 181 men, who were aged 54.7 +/- 14.5 years (mean +/- SD), who had undergone dialysis for 36.3 +/- 33.2 months, were selected randomly from 7 DaVita dialysis units in Los Angeles South/East Bay area. The average weekly dose of administered recombinant human EPO within a 13-week interval was 217 +/- 187 U/kg. Patients were receiving intravenous iron supplementation (iron gluconate or dextran) averaging 39.5 +/- 47.5 mg/wk. The MIS and serum concentrations of high-sensitivity C-reactive protein, interleukin 6 (IL-6), tumor necrosis factor-alpha, and lactate dehydrogenase had positive correlation with required EPO dose and EPO responsiveness index (EPO divided by hemoglobin), whereas serum total iron binding capacity (TIBC), prealbumin and total cholesterol, as well as blood lymphocyte count had statistically significant but negative correlations with indices of refractory anemia. Most correlations remained significant even after multivariate adjustment for case-mix and anemia factors and other relevant covariates. Similar associations were noticed across EPO per body weight tertiles via analysis of variance and after estimating odds ratio for higher versus lower tertile via logistic regression after same case-mix adjustment. CONCLUSION: The existence of elements of MICS as indicated by a high MIS and increased levels of proinflammatory cytokines such as IL-6 as well as decreased nutritional values such as low serum concentrations of total cholesterol, prealbumin, and TIBC correlates with EPO hyporesponsiveness in MHD patients. PMID- 14520628 TI - Rosiglitazone improves glucose metabolism in nondiabetic uremic patients on CAPD. AB - BACKGROUND: Insulin resistance, a strong risk factor for atherosclerotic vascular disease, is present in uremic patients without diabetes on continuous ambulatory peritoneal dialysis (CAPD) therapy. Amelioration of insulin resistance may reduce associated long-term cardiovascular complications. The aim of the study is to investigate the effects of rosiglitazone (ROS), an insulin sensitizer, on glucose metabolism in CAPD patients without diabetes. METHODS: Fifteen uremic patients without diabetes on CAPD therapy were enrolled. All were administered ROS, 4 mg/d, for 12 weeks. A control group consisted of 15 age- and sex-matched healthy subjects. Oral glucose tolerance test (OGTT) results, fasting glucose and insulin levels, related blood biochemistry results, and C-reactive protein (CRP), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha) levels were determined before initiation and at 4 and 12 weeks of therapy. Insulin resistance was evaluated using the homeostasis model assessment method (HOMA-IR). A whole body insulin sensitivity index (ISI) and insulinogenic index for insulin production were calculated from OGTT results. RESULTS: CAPD patients showed significantly greater HOMA-IR and glucose intolerance compared with healthy controls. After 4 and 12 weeks of ROS therapy, there were no significant changes in body weight, blood pressure, dialysis adequacy, hemoglobin level, hemoglobin A(1c) level, liver function, lipid profile, or intact parathyroid hormone, CRP, IL-6, or TNF-alpha levels. There was a significant decrease in HOMA-IR (3.2 +/- 0.6, 2.2 +/- 0.4, and 2.1 +/- 0.4; P < 0.05). During the OGTT, there was a significant decrease in the area under the glucose curve and a significant increase in ISI (3.5 +/- 0.4, 5.0 +/- 0.7, and 5.3 +/- 0.7; P < 0.05), but no significant change in insulinogenic index. CONCLUSION: ROS improved insulin resistance in CAPD patients without diabetes. Whether long-term use of ROS reduces cardiovascular risk needs further study. PMID- 14520629 TI - Association of ENOS polymorphism with basal peritoneal membrane function in uremic patients. AB - BACKGROUND: Basal peritoneal permeability has a major impact on the outcome of peritoneal dialysis (PD) patients, but the determinant of this is unknown. Early evidence suggests that peritoneal permeability is affected by nitric oxide (NO) activity. Recently, a gene polymorphism of the endothelial NO synthase (ENOS) gene was identified that is associated with circulating nitrate levels. METHODS: We performed a cross-sectional study to examine the relationship between ENOS4(a/b) gene polymorphism and basal peritoneal function in 86 Chinese incident PD patients. ENOS genotypes for variable number tandem repeats in intron 4 (a/b) were identified by polymerase chain reaction. Patients were classified into 2 groups according to results of a basal peritoneal equilibration test (PET) performed within 2 months of dialysis therapy: group A consisted of patients with low (L)/L average (LA) PET results, and group B consisted of those with H and HA PET results. RESULTS: Group A (L/LA) had a significantly greater prevalence of ENOS aa/ab genotype than group B (H/HA; 30% versus 12%; P < 0.05). Frequencies of the ENOS a allele also were greater in group A (L/LA) than group B (H/HA) (16% versus 6%; P = 0.03). ENOS genotype remained an independent predictor for peritoneal transport after adjustment for sex, body weight, and prevalence of diabetes by multivariate analysis (adjusted odds ratio, 3.3; confidence interval, 1.1 to 3.7; P = 0.03). Subjects with the aa/ab genotype had significantly lower mass transfer area coefficients (7.35 +/- 3.4 versus 9.48 +/- 5.21 mL/min; P = 0.023) and dialysate-plasma creatinine ratios at 4 hours (0.55 +/- 0.13 versus 0.62 +/- 0.14; P = 0.048) than those with the bb genotype. CONCLUSION: ENOS4(a/b) gene polymorphism is associated with basal peritoneal permeability in uremic Chinese patients. PMID- 14520630 TI - Gastrointestinal and pancreatic function in peritoneal dialysis patients: their relationship with malnutrition and peritoneal membrane abnormalities. AB - BACKGROUND: Malnutrition is frequent in peritoneal dialysis (PD) patients, but the contribution of gastrointestinal (GI) dysfunction has not been well established. METHODS: We studied GI function in 49 stable PD patients to ascertain its relationship with malnutrition. After an overload fat diet, fecal fat, sugar, starch and nitrogen, intestinal protein permeability (alpha(1) antitrypsin fecal clearance [C-alpha(1)-AT]), fecal chymotrypsin (CT), GI hormones and gastrin, pepsinogen I and II, cholecystokinin (CCK), gastrin releasing peptide (GRP), and neuropeptide Y (NPY) were measured. Vasoactive intestinal polypeptide (VIP), substance P (SP), and tumor necrosis factor (TNF alpha) and biochemical nutritional markers were evaluated. RESULTS: All patients showed high fecal sugar. Elevated fecal nitrogen was found in 21 patients, 6 with high C-alpha(1)-AT. High fecal starch levels appeared in 21, fat in 20, and low fecal CT in 39 patients. These determinations showed inverse relation with nutritional markers. Increased fecal C-alpha(1)-AT values were associated with lower serum albumin. Fecal CT values showed a negative linear correlation with serum albumin and were inversely associated with retinol-binding protein, normalized protein nitrogen appearance, and serum iron. High plasma levels of pancreatic stimulating hormones were found: gastrin, CCK, and VIP. These levels were higher in patients with a worse pancreatic exocrine function. Higher values of other GI hormones, gastrin, pepsinogen I and II, CCK, GRP, and TNF-alpha. Normal concentrations of NPY, VIP, and PS were observed. CONCLUSION: GI abnormalities (malabsorption, maldigestion, pancreatic dysfunction, and protein losing enteropathy) are present in an important number of PD patients. These features are negatively associated to nutrition. PMID- 14520631 TI - Hypercoagulability, a serious problem in patients with ESRD on maintenance hemodialysis, and its correction after kidney transplantation. AB - BACKGROUND: Recurrent vascular access thrombosis (VAT) resulting in failure to continue maintenance hemodialysis (HD) therapy is not an uncommon event. The cause of VAT in these circumstances remains uncertain. We describe results of our studies to identify changes in hemostatic balance in patients on maintenance HD therapy that probably contributed to a hypercoagulable state. METHODS: We studied 82 patients with end-stage renal disease on maintenance HD therapy who underwent HD for 11 to 52 months (39.3 +/- 27.4 months). Forty-nine episodes of VAT occurred in 22 patients; a single episode occurred in 12 patients; and 2 or more episodes, in 10 patients. Blood coagulation studies, including assays of inhibitors and activated protein C (PC) resistance (APCR), were performed using standard techniques. RESULTS: Investigations showed the presence of lupus anticoagulant (LA) in 5.6%, anticardiolipin antibody immunoglobulin G (IgG) in 3.9% and IgM in 5.3%, APCR in 20.5%, and deficiencies in protein S (PS), PC, and antithrombin III (ATIII) in 32.1%, 24.4%, and 19.2%, respectively. When parameters were compared between patients with and without VAT episodes, LA, PC, PS, and APCR levels were significantly abnormal in those who experienced VAT. Sixteen subjects with hypercoagulable states on HD therapy underwent renal transplantation and were evaluated 9.3 +/- 4.2 months posttransplantation. Deficiencies in PC (P = 0.014), PS (P = 0.001), ATIII (P = 0.017), and APCR (P = 0.0001) were completely corrected in all subjects. CONCLUSION: Hypercoagulability is a risk factor for recurrent VAT in HD patients, and renal transplantation successfully corrects these abnormalities. PMID- 14520632 TI - Developing clinical performance measures based on the Dialysis Outcomes Quality Initiative Clinical Practice Guidelines: process, outcomes, and implications. AB - BACKGROUND: The National Kidney Foundation-Dialysis Outcomes Quality Initiative (NKF-DOQI) Clinical Practice Guidelines established a widely accepted set of recommendations for high-quality dialysis care. To enhance the End-Stage Renal Disease Core Indicators Project, an ongoing effort to assess and improve dialysis care in the United States, the Centers for Medicare and Medicaid Services (CMS) commissioned a project to develop clinical performance measures (CPMs) based on the NKF-DOQI guidelines. METHODS: The CMS contracted with Qualis Health, a private nonprofit organization serving as a Medicare Quality Improvement Organization, to facilitate a 9-month project to develop dialysis CPMs with the participation of a broad range of stakeholders from the renal community. Work groups were established to develop CPMs addressing 4 areas: hemodialysis adequacy, peritoneal dialysis adequacy, vascular access management, and anemia management. The NKF-DOQI guidelines were prioritized based on the strength of the evidence supporting the guidelines, the feasibility of developing performance measures, and the significance of the areas addressed to the quality of care delivered to dialysis patients. Expert panels developed data specifications, sampling approaches, data-collection tools, and analytic strategies. RESULTS: Sixteen CPMs were developed based on 22 of 114 NKF-DOQI guidelines. After establishing reliability through field-testing of data-collection instruments, the CPMs were applied to a sample of 8,838 randomly selected hemodialysis patients and 1,650 randomly selected adult peritoneal dialysis patients in summer 1999. CONCLUSION: The development of CPMs based on the NKF-DOQI Clinical Practice Guidelines for dialysis care was accomplished in a timely and effective manner by engaging a broad range of stakeholders and technical experts. The CPMs are important tools to assess and improve the quality of dialysis care in the United States. Few comparable efforts exist in other fields of medicine. PMID- 14520633 TI - The need for end-of-life care training in nephrology: national survey results of nephrology fellows. AB - Because of the high mortality rate of end-stage renal disease, nephrologists care for many dying patients. However, the education of nephrology fellows in palliative care has not been assessed. We surveyed second-year nephrology fellows to assess the quantity and quality of teaching they received in palliative medicine and also asked about their preparedness to manage patients at the end of life. A 63% survey response rate yielded 173 surveys for evaluation. Nearly all fellows (99%) agreed that physicians have a responsibility to help patients at the end of life; half thought it was very important to learn how to care for dying patients. On a 10-point scale in which 0 is no teaching and 10 is a lot of teaching, fellows reported significantly less teaching in end-of-life care (mean score, 3.8 +/- 2.6) than in managing a patient with distal renal tubular acidosis (mean score, 6.3 +/- 2.5) or on hemodialysis therapy (mean score, 8.9 +/- 1.5; all P < 0.0001). Specific palliative care content areas were taught infrequently; only 22% of fellows were taught how to tell a patient he or she is dying. Fellows who had contact with a palliative care specialist reported more education on end of-life issues and believed they were better prepared to provide such care. Fellows' palliative care experiences during fellowship frequently occurred without attending nephrologist supervision; 32% of fellows had conducted 2 or fewer family meetings, and 26% of all family meetings occurred without an attending nephrologist. Fellows believed they were best prepared to manage a patient on hemodialysis therapy (mean score, 8.9 +/- 1) and least prepared to manage a patient at the end of life (mean score, 6.1 +/- 2; P < 0.0001). Our results show that most nephrology fellows believe they should learn how to care for dying patients, but most fellowship programs do not offer this training. Our study therefore suggests that training in palliative care be incorporated into fellowship program curricula. PMID- 14520634 TI - Retransplantation after kidney graft loss due to polyoma BK virus nephropathy: successful outcome without original allograft nephrectomy. AB - Although polyoma BK virus (BKV)-associated interstitial nephritis has received increasing attention because of its clinical relevance in kidney allograft recipients, data on risk for repeated renal transplantation after BKV-related allograft loss are limited, and the need to perform an original graft nephrectomy is the object of debate. A 15-year-old boy with renal failure secondary to Alport's syndrome underwent renal transplantation. His posttransplantation course was complicated by acute rejection episodes and the presence of circulating anti glomerular basement membrane antibodies that required aggressive immunosuppressive treatment. Graft failure caused by BKV-associated interstitial nephropathy occurred despite a reduction in immunosuppression and cidofovir treatment. The patient received a second transplant without an original graft nephrectomy, and 15 months after retransplantation, he persists with optimal graft function and is constantly BKV DNA negative in both urine and plasma. Our report indicates that an original allograft nephrectomy may not be mandatory for successful retransplantation after graft loss caused by BKV nephropathy. PMID- 14520635 TI - Approach to renal biopsy. PMID- 14520637 TI - Rapidly progressive irreversible renal failure in patients with pancreatic insufficiency. PMID- 14520639 TI - Vitamin C for hyporesponsiveness to EPO: a cure for all? PMID- 14520647 TI - Recessive type of nephrogenic diabetes insipidus. PMID- 14520649 TI - Recurrence of ANCA-positive glomerulonephritis immediately after renal transplantation. AB - Recurrence of crescentic necrotizing glomerulonephritis after renal transplantation is rare. Successful renal transplantation in patients with antineutrophil cytoplasmic autoantibody (ANCA) glomerulonephritis has been reported. The presence of ANCA at transplantation does not appear to increase the rate of relapse after kidney allografting. Therapy with cyclophosphamide and corticoids usually is effective. We report a case of recurrent perinuclear ANCA crescentic necrotizing glomerulonephritis immediately after renal transplantation that was treated successfully by cyclophosphamide, plasma exchange, and intravenous polyvalent immunoglobulin. PMID- 14520650 TI - The cognitive consequences of epilepsy. PMID- 14520651 TI - Paternal transmission of mitochondrial DNA is (fortunately) rare. PMID- 14520652 TI - Chronic epilepsy and cognition: a longitudinal study in temporal lobe epilepsy. AB - It remains unclear whether uncontrolled epilepsy causes mental decline. This longitudinal study contrasts change of memory and nonmemory functions in 147 surgically and 102 medically treated patients with temporal lobe epilepsy. All participants were evaluated at baseline (T1) and after 2 to 10 years (T3). Surgical patients underwent additional testing 1 year postoperatively (T2). Data were analyzed on an individual and group level. Sixty-three percent of the surgical and 12% of the medically treated patients were seizure-free at T3. Fifty percent of the medically treated and 60% of the surgical patients showed significant memory decline at T3 with little change in nonmemory functions (difference not significant). Surgery anticipated the decline seen in the medically treated group and exceeded it when surgery was performed on the left, or if seizures continued postoperatively. Seizure-free surgical patients showed recovery of nonmemory functions at T2 (p < 0.001) and of memory functions at T3 (T3, p = 0.03). Multiple regression indicated retest interval, seizure control, and mental reserve capacity as predictors of performance changes. In addition, psychosocial outcome was better when seizures were controlled. In conclusion, chronic temporal lobe epilepsy is associated with progressive memory impairment. Surgery, particularly if unsuccessful, accelerates this decline. However, memory decline may be stopped and even reversed if seizures are fully controlled. PMID- 14520653 TI - Onset of dementia is associated with age at menopause in women with Down's syndrome. AB - Women with Down's syndrome experience early onset of both menopause and Alzheimer's disease. This timing provides an opportunity to examine the influence of endogenous estrogen deficiency, indicated by age at menopause, on risk of Alzheimer's disease. A community-based sample of 163 postmenopausal women with Down's syndrome, 40 to 60 years of age, was ascertained through the New York State Developmental Disability service system. Information from cognitive assessments, medical record review, neurological evaluation, and caregiver interviews was used to establish ages for onset of menopause and dementia. We used survival and multivariate regression analyses to determine the relation of age at menopause to age at onset of Alzheimer's disease, adjusting for age, level of mental retardation, body mass index, and history of hypothyroidism or depression. Women with early onset of menopause (46 years or younger) had earlier onset and increased risk of Alzheimer's disease (AD) compared with women with onset of menopause after 46 years (rate ratio, 2.7; 95% confidence interval [CI], 1.2-5.9). Demented women had higher mean serum sex hormone binding globulin levels than nondemented women (86.4 vs 56.6 nmol/L, p = 0.02), but similar levels of total estradiol, suggesting that bioavailable estradiol, rather than total estradiol, is associated with dementia. Our findings support the hypothesis that reductions in estrogens after menopause contribute to the cascade of pathological processes leading to AD. PMID- 14520654 TI - Early recovery after cerebral ischemia risk of subsequent neurological deterioration. AB - Given the high short-term risk of stroke after transient ischemic attack, we hypothesized that substantial acute neurological recovery in patients presenting with cerebral ischemia would be associated with a greater risk of subsequent neurological deterioration due to recurrent cerebral ischemia. Data from the Trial of ORG10172 in Acute Stroke Treatment, a randomized trial of the heparinoid danaparoid, were analyzed to determine whether substantial acute recovery, defined as an improvement of greater than or equal to 75% on National Institutes of Health Stroke Scale (NIHSS) between baseline and 24 hours, was associated with a greater risk of subsequent deterioration, defined as a worsening on the NIHSS between day 1 and day 90. Of 1,184 subjects meeting entry criteria, 63 (5.3%) had substantial acute recovery. Subsequent deterioration was more common in those with substantial acute recovery compared with others (48 vs 33%; p = 0.028 by Fisher's exact test). In multivariable models, substantial acute recovery remained an independent predictor of subsequent deterioration (odds ratio, 3.0; 95% confidence interval, 1.7-5.2; p < 0.001). Among patients with acute cerebral ischemia, those who recover substantially within 24 hours may be at greater risk of subsequent neurological deterioration due to causes other than hemorrhage. PMID- 14520655 TI - Subthalamic nucleus stimulation affects a frontotemporal network: a PET study. AB - Deep brain stimulation (DBS) of the subthalamic nucleus (STN) has become an effective strategy in the treatment of motor symptoms in advanced Parkinson's disease. However, clinical studies have shown that DBS can affect verbal fluency. Seven Parkinson's disease patients with bilateral DBS of the STN were studied with positron emission tomography (PET) to investigate the effects of STN stimulation on regional cerebral blood flow during a verbal fluency task. Activation of the right orbitofrontal cortex and verbal fluency-associated activation within a left-sided frontotemporal network were decreased during STN stimulation compared with the OFF state. Our results offer an explanation for the commonest neuropsychological side effect of STN stimulation and show that STN stimulation affects a frontotemporal network during a fluency task. PMID- 14520656 TI - Meningitis-associated hearing loss: protection by adjunctive antioxidant therapy. AB - Hearing loss is the most frequent long-term complication of pneumococcal meningitis, affecting up to 40% of survivors. Unfortunately, adjuvant therapy with dexamethasone has failed to satisfactorily reduce its incidence. Therefore, we evaluated the use of antioxidants for the adjunctive therapy of meningitis associated deafness. Eighteen hours after intracisternal injection of 7.5 x 10(5) colony-forming units of Streptococcus pneumoniae, rats were treated systemically either with ceftriaxone and the antioxidants and peroxynitrite scavengers Mn(III)tetrakis(4-benzoic acid)-porphyrin (MnTBAP) or N-acetyl-L-cysteine (NAC) or placebo (1 ml phosphate-buffered saline) for 4 days. Hearing was assessed by auditory brainstem response audiometry. Adjunctive antioxidant therapy significantly reduced the long-term hearing loss (14 days after infection) for square wave impulses (mean hearing loss +/- SD: ceftriaxone and placebo, 45+/-26 dB; ceftriaxone and MnTBAP, 9+/-23 dB; ceftriaxone and NAC, 19+/-30 dB) as well as 1 kHz (ceftriaxone and placebo, 28+/-19 dB; ceftriaxone and MnTBAP, 10+/-16 dB; ceftriaxone and NAC, 10+/-17 dB), and 10 kHz tone bursts (ceftriaxone and placebo, 62+/-27 dB; ceftriaxone and MnTBAP, 16+/-13 dB; ceftriaxone and NAC, 25+/-26 dB). Furthermore, both antioxidants attenuated the morphological correlates of meningogenic hearing loss, namely, long-term blood-labyrinth barrier disruption, spiral ganglion neuronal loss, and fibrous obliteration of the perilymphatic spaces. Adjuvant antioxidant therapy is highly otoprotective in meningitis and therefore is a promising future treatment option. PMID- 14520657 TI - Oligoclonal immunoglobulins in cerebrospinal fluid during varicella zoster virus (VZV) vasculopathy are directed against VZV. AB - Limited analyses of cerebrospinal fluid from patients with central nervous system infections have shown that the oligoclonal IgG is antibody directed against the agent that causes disease. Using a new method involving binding of IgG to beads coated with lysates prepared from candidate infectious antigens, we showed that the oligoclonal IgG in cerebrospinal fluid of a patient with chronic varicella zoster virus vasculopathy is directed against the causative virus. This approach holds promise in identifying and purifying the relevant oligoclonal IgGs in inflammatory central nervous system diseases of unknown cause. PMID- 14520658 TI - Contribution of the ipsilateral motor cortex to recovery after chronic stroke. AB - It has been proposed that the intact (ipsilateral) motor cortex play a significant role mediating recovery of motor function in the paretic hand of chronic stroke patients, but this hypothesis has not been tested experimentally. Here, we evaluated the effects of transcranial magnetic stimulation (TMS) on motor performance of the paretic hand of chronic stroke patients and healthy controls. We hypothesized that, if activity in the intact hemisphere contributes to functional recovery, TMS should result in abnormal motor behavior in the paretic hand. We found that stimulation of the intact hemisphere resulted in delayed simple reaction times (RTs) in the contralateral healthy but not in the ipsilateral paretic hand, whereas stimulation of the lesioned hemisphere led to a marked delay in RT in the contralateral paretic hand but not in the ipsilateral healthy hand. RT delays in the paretic hand correlated well with functional recovery. Finger tapping in the paretic hand was affected by TMS of the lesioned but not the intact hemisphere. These results are consistent with the idea that recovered motor function in the paretic hand of chronic stroke patients relies predominantly on reorganized activity within motor areas of the affected hemisphere. PMID- 14520659 TI - Low mutant load of mitochondrial DNA G13513A mutation can cause Leigh's disease. AB - Respiratory chain complex I deficiency is a common cause of Leigh's disease (LD) and can be caused by mutations in genes encoded by either nuclear or mitochondrial DNA (mtDNA). Most pathogenic mtDNA mutations act recessively and only cause disease when present at high mutant loads (typically >90%) in tissues such as muscle and brain. Two mitochondrial DNA mutations in complex I subunit genes, G14459A in ND6, and T12706C in ND5, have been associated with complex I deficiency and LD. We report another ND5 mutation, G13513A, in three unrelated patients with complex I deficiency and LD. The G13513A mutation was present at mutant loads of approximately 50% or less in all tissues tested, including multiple brain regions. The threshold mutant load for causing a complex I defect in cultured cells was approximately 30%. Blue Native polyacrylamide gel electrophoresis showed that fibroblasts with 45% G13513A mutant load had approximately 50% of the normal amount of fully assembled complex I. Fibroblasts with greater than 97% of the ND6 G14459A mutation had only 20% fully assembled complex I, suggesting that both mutations disrupt complex I assembly or turnover. We conclude that the G13513A mutation causes a complex I defect when present at unusually low mutant load and may act dominantly. PMID- 14520660 TI - Imaging-guided convection-enhanced delivery and gene therapy of glioblastoma. AB - In a prospective phase I/II clinical study, we treated eight patients suffering from recurrent glioblastoma multiform with stereotactically guided intratumoral convection-enhanced delivery of an HSV-1-tk gene-bearing liposomal vector and systemic ganciclovir. Noninvasive identification of target tissue together with assessment of vector-distribution volume and the effects of gene therapy were achieved using magnetic resonance imaging and positron emission tomography. The treatment was tolerated well without major side effects. In two of eight patients, we observed a greater than 50% reduction of tumor volume and in six of eight patients focal treatment effects. Intracerebral infusion of contrast medium before vector application displayed substantial inhomogeneity of tissue staining indicating the need of test infusions to monitor the mechanical distribution of vectors. Visualization of therapeutic effects on tumor metabolism and documentation of gene expression using positron emission tomography indicated that molecular imaging technology appears to be essential for the further development of biological treatment strategies. PMID- 14520661 TI - Predictors of neuropathological severity in 100 patients with Huntington's disease. AB - Subjects were followed in the Longitudinal Core Study of the Baltimore Huntington's Disease Center and given annual neurological, cognitive, and psychiatric examinations. Postmortem neuropathological grade was assigned using the system of Vonsattel and colleagues. We examined the correlations between the neuropathological grade and scores on the Quantified Neurological Examination (QNE) and its chorea and motor impairment subscales, the Mini-Mental State Examination (MMSE), the HD Activities of Daily Living (ADL) Scale, and a number of demographic variables (CAG number, age of onset, age at death, disease duration) in 100 subjects who had been examined within 1,000 days of death. All measures showed significant correlation with Vonsattel score except for chorea. The strongest effect was that of motor impairment score (r(2) = 0.351, p < 0.0001). In a stepwise correlation of clinical variables, motor impairment remained significant. The largest effect for a demographic variable was for age of onset (r(2) = 0.226, p < 0.0001) A partial correlation was significant between CAG number and Vonsattel grade, controlling for age at death or disease duration. Motor impairment appears to be a good clinical measure of neuronal cell loss, at least late in the course of HD and therefore may prove useful in observational and treatment studies. PMID- 14520662 TI - Myosin storage myopathy associated with a heterozygous missense mutation in MYH7. AB - Myosin constitutes the major part of the thick filaments in the contractile apparatus of striated muscle. MYH7 encodes the slow/beta-cardiac myosin heavy chain (MyHC), which is the main MyHC isoform in slow, oxidative, type 1 muscle fibers of skeletal muscle. It is also the major MyHC isoform of cardiac ventricles. Numerous missense mutations in the globular head of slow/beta-cardiac MyHC are associated with familial hypertrophic cardiomyopathy. We identified a missense mutation, Arg1845Trp, in the rod region of slow/beta-cardiac MyHC in patients with a skeletal myopathy from two different families. The myopathy was characterized by muscle weakness and wasting with onset in childhood and slow progression, but no overt cardiomyopathy. Slow, oxidative, type 1 muscle fibers showed large inclusions consisting of slow/beta-cardiac MyHC. The features were similar to a previously described entity: hyaline body myopathy. Our findings indicate that the mutated residue of slow/beta-cardiac MyHC is essential for the assembly of thick filaments in skeletal muscle. We propose the term myosin storage myopathy for this disease. PMID- 14520663 TI - Microdysgenesis in mesial temporal lobe epilepsy: a clinicopathological study. AB - The interrelationship of mesial temporal lobe epilepsy (MTLE), hippocampal sclerosis, and febrile convulsions still remains an enigma. Additional microscopical cortical dysplasia or microdysgenesis has been suggested as pre existent susceptibility factor rendering the affected brain vulnerable to the development of MTLE after initial precipitating injuries such as febrile convulsions. Twenty-four MTLE cases with histopathologically definite hippocampal sclerosis were examined for clearly defined features of microdysgenesis and further signs of neocortical dysplasia. Although unequivocal signs of dysplasia were absent, 29.2% of cases showed cortical neuronal clustering, 25.0% showed perivascular clustering, and 20.8% showed increased white matter neurons. The features of microdysgenesis studied here were not linked with each other and were not related to initial precipitating injuries, positive family history, or any other clinical parameter. Their suggested fundamental role as dysplastic factor within development of hippocampal sclerosis and MTLE is not confirmed. PMID- 14520664 TI - Familial aggregation of early- and late-onset Parkinson's disease. AB - The role of heredity in early- versus late-onset Parkinson's disease (PD) is controversial. We estimated the degree of increased risk of PD in first-degree relatives of 221 PD probands with age of onset 50 years or younger and 266 PD probands with age of onset older than 50 years compared with the first-degree relatives of 409 control probands. Risk of PD was similar among first-degree relatives of early-onset PD probands (risk ratio [RR], 2.9; 95% confidence interval [CI], 1.6-5.0; p = 0.0002) and late-onset PD probands (RR, 2.7; 95% CI, 1.6-4.4; p = 0.0002) when each was compared with first-degree relatives of controls. However, siblings of early-onset PD probands were at markedly increased risk of PD compared with siblings of controls (RR, 7.9; 95% CI, 2.5-25.5; p = 0.0005), whereas parents of early-onset PD probands were not at significantly increased risk compared with parents of controls (RR, 1.7; 95% CI, 0.9-3.3; p = 0.2). In late-onset families, both siblings (RR, 3.6; 95% CI, 1.3-10.3; p = 0.02) and parents (RR, 2.5; 95% CI, 1.4-4.6; p = 0.003) were at increased risk compared with control relatives. This pattern is consistent with an autosomal recessive contribution to the inheritance of early but not late-onset PD. Genetic factors are important in both early- and late-onset PD, but specific genes and mode of inheritance may differ between the two groups. PMID- 14520665 TI - Epilepsy in low-grade gliomas: the impact on cognitive function and quality of life. AB - Low-grade gliomas frequently are associated with epilepsy. The purpose of this study is to determine the impact of epilepsy and antiepileptic drug (AED) treatment on cognitive functioning and health-related quality of life (HRQOL) in these patients. One hundred fifty-six patients without clinical or radiological signs of tumor recurrence for at least 1 year after histological diagnosis and with an epilepsy burden (based on seizure frequency and AED use) ranging from none to severe were compared with healthy controls. The association between epilepsy burden and cognition/HRQOL was also investigated. Eighty-six percent of the patients had epilepsy and 50% of those using AEDs actually were seizure-free. Compared with healthy controls, glioma patients had significant reductions in information processing speed, psychomotor function, attentional functioning, verbal and working memory, executive functioning, and HRQOL. The increase in epilepsy burden that was associated with significant reductions in all cognitive domains except for attentional and memory functioning could primarily be attributed to the use of AEDs, whereas the decline in HRQOL could be ascribed to the lack of complete seizure control. In conclusion, low-grade glioma patients suffer from a number of neuropsychological and psychological problems that are aggravated by the severity of epilepsy and by the intensity of the treatment. PMID- 14520666 TI - Genotypes from patients indicate no paternal mitochondrial DNA contribution. AB - A cornerstone of mitochondrial genetics, strict maternal inheritance, has been challenged recently by the study of a patient with mitochondrial myopathy due to a sporadic 2bp deletion. The mitochondrial DNA (mtDNA) harboring the mutation was paternal in origin, whereas the patient's blood was identical to the maternal genotype. To determine whether this is a common phenomenon, we studied mtDNA sequence variation between muscle and blood from 35 patients with sporadic mitochondrial myopathies, but detected no evidence of paternal mtDNA transmission. Our findings suggest that paternal transmission of mtDNA is rare and should not alter our genetic advice to families. PMID- 14520667 TI - Lack of paternal inheritance of muscle mitochondrial DNA in sporadic mitochondrial myopathies. AB - In 2002, paternal inheritance of muscle mitochondrial DNA (mtDNA) was reported in a patient with exercise intolerance and a mitochondrial DNA (mtDNA) mutation restricted to skeletal muscle. To evaluate whether paternal inheritance is a common phenomenon, we studied 10 sporadic patients with skeletal muscle restricted mtDNA mutations: five harbored mtDNA point mutations in protein-coding genes and five had single mtDNA deletions. We performed haplotype analysis and direct sequencing of the hypervariable regions 1 and 2 of the D-loop in muscle and blood from the patients and, when available, in blood from their parents. We did not observe paternal inheritance in any of our patients. PMID- 14520668 TI - Bilateral striatal necrosis associated with a novel mutation in the mitochondrial ND6 gene. AB - We report the molecular findings in two independent patients presenting with progressive generalized dystonia and bilateral striatal necrosis in whom we have identified a mutation (T14487C) in the mitochondrial ND6 gene. The mutation is heteroplasmic in all samples analyzed, and it fulfills all accepted criteria of pathogenicity. Transmitochondrial cell lines harboring 100% mutant mitochondrial DNA showed a marked decrease in the activity of complex I of the respiratory chain supporting the pathogenic role of T14487C. PMID- 14520669 TI - Voltage-gated potassium channel antibodies in limbic encephalitis. AB - We found voltage-gated potassium channel (VGKC) antibodies in 4 of 15 patients with limbic encephalitis (LE). Two patients with idiopathic LE had high VGKC antibody levels (>800 pM; controls <100 pM), that fell in parallel with a clinical response to immunotherapy. Two patients with lower VGKC antibodies (170 pM, 300 pM) had lung cancer (radiological evidence only in one) and the LE improved with immunotherapy in one. The other 11 patients without VGKC antibodies had paraneoplastic LE and eight onconeural antibodies (Hu in 6; Ma2 in 2). VGKC antibodies do not unambiguously discriminate between idiopathic or paraneoplastic LE but probably indicate a good response to immunotherapy. PMID- 14520670 TI - Retrograde reactions of Clarke's nucleus neurons after human spinal cord injury. AB - Successful axon regeneration depends on the expression of regeneration-associated genes by axotomized neurons. Here, we demonstrate, for the first time to our knowledge, the expression of regeneration-associated genes by axotomized human CNS neurons. In situ hybridization and immunohistochemistry showed a transient induction of GAP-43 and c-jun in Clarke's nucleus neurons caudal to traumatic human spinal cord injury. These results support experimental data that nonregenerating central nervous system neurons can temporarily upregulate regeneration-associated genes, reflecting a transient regenerative capacity that fails over time. PMID- 14520671 TI - A diagnostic cycle test for McArdle's disease. AB - We investigated whether the second wind phenomenon (ie, a decrease in heart rate and perceived exertion during exercise) is pathognomonic for McArdle's disease. Twenty-four patients with McArdle's disease, 17 healthy subjects, and 25 patients with other inborn errors of muscle metabolism cycled a constant workload for 15 minutes. In McArdle's disease patients, heart rate consistently decreased by 35 +/- 3 beats per minute from the 7(th) to the 15(th) minute of exercise, whereas heart rate increased progressively with exercise in all 42 control subjects. The findings indicate that cycling at a moderate, constant workload provides a specific, sensitive, and simple diagnostic test for McArdle's disease. PMID- 14520672 TI - Retinal involvement in dementia with Lewy bodies: a clue to hallucinations? AB - Visual hallucinations are a core feature of dementia with Lewy bodies. Their pathophysiology is not well understood, because neither clinical nor histological data have shown their basic mechanisms. Here, we report the presence of pale inclusions in the outer plexiform layer of the retina in a patient with dementia with Lewy bodies. These inclusions are related to cytoskeletal disorganization of the cones at ultrastructural level and modifications of the immunohistochemical pattern of distribution of synucleins in the retina. These modifications may participate in the visual impairment in dementia with Lewy bodies. PMID- 14520673 TI - Neuropathological findings in West Nile virus encephalitis: a case report. AB - A 67-year-old woman had fever, myalgias, progressive weakness, and respiratory insufficiency. In 9 days, flaccid areflexic quadriparesis and bulbar palsy developed. She died 26 days after the onset of her illness. Serum and cerebrospinal fluid serology were positive for West Nile virus. Neuropathological study showed changes consistent with a viral encephalomyelitis, similar to poliomyelitis. The brainstem showed neuronal loss and multiple foci of necrosis. The spinal cord showed severe loss of anterior and posterior horn neurons. Immunohistochemistry identified West Nile virus antigens in the brainstem and spinal cord. Paralysis, in West Nile virus encephalitis, is caused by destruction of motor neurons. PMID- 14520674 TI - Cardiac bioenergetics in Friedreich's ataxia. PMID- 14520678 TI - Is decoupling of autonomic and cognitive emotional reactions a manifestation of cerebellar stroke or hydrocephalus? PMID- 14520676 TI - Absence of association between codon 129/219 polymorphisms of the prion protein gene and Alzheimer's disease in Japan. PMID- 14520682 TI - Pharmacokinetics and pharmacodynamics of azosemide. AB - Azosemide is used in the treatment of oedematous states and hypertension. The exact mechanism of action is not fully understood, but it mainly acts on both the medullary and cortical segments of the thick ascending limb of the loop of Henle. Delayed tolerance was demonstrated in humans by homeostatic mechanisms (principally an increase in aldosterone secretion and perhaps also an increase in the reabsorption of solute in the proximal tubule). After oral administration to healthy humans in the fasting state, the plasma concentration of azosemide reached its peak at 3-4 h with an absorption lag time of approximately 1 h and a terminal half-life of 2-3 h. The estimated extent of absolute oral bioavailability in humans was approximately 20.4%. After oral administration of the same dose of azosemide and furosemide, the diuretic effect was similar between the two drugs, but after intravenous administration, the effect of azosemide was 5.5-8 times greater than that in furosemide. This could be due to the considerable first-pass effect of azosemide. The protein binding to 4% human serum albumin was greater than 95% at azosemide concentrations ranging from 10 to 100 microg/ml using an equilibrium dialysis technique. The poor affinity of human tissues to azosemide was supported by the relatively small value of the apparent post-pseudodistribution volume of distribution (Vdbeta), 0.262 l/kg. Eleven metabolites (including degraded products) of azosemide including M1, glucuronide conjugates of both M1 and azosemide, thiophenemethanol, thiophencarboxylic acid and its glycine conjugate were obtained in rats. Only azosemide and its glucuronide were detected in humans. In humans, total body clearance, renal clearance and terminal half-life of azosemide were 112 ml/min, 41.6 ml/min and 2.03 h, respectively. Azosemide is actively secreted in the renal proximal tubule possibly via nonspecific organic acid secretory pathway in humans. Thus, the amount of azosemide that reaches its site of action could be significantly modified by changes in the capacity of this transport system. This capacity, in turn, could be predictably changed in disease states, resulting in decreased delivery of the diuretic to the transport site, as well as in the presence of other organic acids such as nonsteroidal anti-inflammatory drugs which could compete for active transport of azosemide. The urinary excretion rate of azosemide could be correlated well to its diuretic effects since the receptors are located in the loop of Henle. The diuretic effects of azosemide were dependent on the rate and composition of fluid replacement in rabbits; therefore, this factor should be considered in the evaluation of bioequivalence assessment. PMID- 14520683 TI - Comparison of ceftibuten transport across Caco-2 cells and rat jejunum mounted on modified Ussing chambers. AB - Ceftibuten uptake into Caco-2 cells and intestinal brush border membrane vesicles is mediated by the dipeptide transport system (PEPT1). The apical to basolateral transport characteristics of ceftibuten across Caco-2 cells and rat jejunum mounted on a modified Ussing chamber was examined. Mannitol was used as a paracellular marker along with trans-epithelial electrical resistance (TEER) for monitoring tight junction permeability. Transport across Caco-2 cells and rat jejunum mounted on a modified Ussing chamber was linear across the concentration range 0.25-10 mM. The net flux of mannitol and ceftibuten was higher across rat jejunum compared with Caco-2 cells. At a donor concentration of 0.25 mM, ceftibuten transport across Caco-2 cells was found to be pH dependent. Glycyl proline, a dipeptide, and 2,4- dinitrophenol, an energy poison, caused a reduction in the permeability of 0.25 mM ceftibuten across Caco-2 cells. Benzoic acid and adipic acid also inhibited transcellular transport of ceftibuten. At a donor concentration of 0.25 mM, passive paracellular transport accounts for about 60% and the active carrier mediated mechanism accounts for about 40% of ceftibuten transport across Caco-2 cells. None of the inhibitors however, had a significant effect on ceftibuten transport across rat jejunum mounted on a modified Ussing chamber at a donor concentration of 0.25 mM. In the concentration range 0.25-10 mM, ceftibuten is predominantly transported by paracellular mechanisms across rat jejunum and a mixture of active and passive transport across Caco-2 cells. PMID- 14520684 TI - Development of the second-order derivative UV spectrophotometric method for direct determination of paracetamol in urine intended for biopharmaceutical characterisation of drug products. AB - Paracetamol is a widely used nonsalicylate analgesic and antipyretic drug. The existing methods for the determination of paracetamol in biological fluids are mainly HPLC techniques, although there are some reported methods based on spectrophotometric determinations. However, all these methods involve some extraction or derivatisation procedures. In the present study the UV spectra of investigated samples were recorded over the wavelength range 220-400 nm (lambda step 0.21 nm; scan speed 60 nm/min) and second-order derivative spectra were calculated. Second-order derivative spectra of different blank urine samples displayed the presence of a zero-crossing point at 245-247 nm defined as lambdazc. The zero-order absorption spectra of paracetamol in water displays maximum absorbance at 243 nm, while in second derivative spectra, a minimum peak at 246 nm was observed. Therefore, the application of zero-crossing technique to the second-derivative UV absorption spectrum should be useful for the determination of paracetamol using 2Dlambdazc. The proposed method enables determination of total paracetamol in urine directly and simply by reading the 2Dlambdazc of the diluted samples. The obtained results were in good accordance with published data on cumulative urinary excretion after per oral administration of paracetamol obtained applying different spectrophotometric methods of determination. It could be useful for biopharmaceutical characterisation of drug products (monitoring of the levels of paracetamol in urine in bioavailability testing, for the evaluation of in vitro-in vivo correlation and screening of different formulations during drug product development). PMID- 14520685 TI - Bioequivalence evaluation of two brands of enalapril 20 mg tablets (Narapril and Renitec) in healthy human volunteers. AB - The bioequivalence of two brands of enalapril 20 mg tablets was demonstrated in 24 healthy human volunteers after a single oral dose in a randomized cross-over study, conducted at IPRC, Amman, Jordan. Reference (Renitec, MSD, Netherlands) and test (Narapril, Julphar, UAE) products were administered to fasted male volunteers; blood samples were collected at specified time intervals, plasma separated and analysed for enalapril and its active metabolite (enalaprilat) using a validated LC-MS/MS method at Cartesius Analytical Unit, Institute of Biomedical Sciences, USP, Sao Paulo, Brazil. The pharmacokinetic parameters AUC(0 t), AUC(0-infinity), Cmax, Tmax, T(1/2) and elimination rate constant were determined from plasma concentration-time profile for both formulations and were compared statistically to evaluate bioequivalence between the two brands, using the statistical modules recommended by FDA. The analysis of variance (ANOVA) did not show any significant difference between the two formulations and 90% confidence intervals fell within the acceptable range for bioequivalence. Based on these statistical inferences it was concluded that the two brands exhibited comparable pharmacokinetic profiles and that Julphar's Narapril is bioequivalent to Renitec of MSD, Netherlands. PMID- 14520686 TI - Relaxin-like peptides in cancer. AB - The members of the relaxin-like hormone family, relaxin and INSL3, also known as relaxin-like factor (RLF) or Leydig cell-derived insulin-like factor (LEY-I-L), are implicated in various mechanisms associated with tumor cell growth, differentiation, invasion and neovascularization. The recent discovery of the relaxin receptor LGR7 and the INSL3/relaxin receptor LGR8 has provided evidence of an auto/paracrine relaxin-like action in tumor tissues and enables the elucidation of the cellular pathways involved in the proposed functions of relaxin in tumor biology. Our review summarizes our current knowledge of the expression of relaxin and INSL3 in human neoplastic tissues and discusses the etiological roles of these heterodimeric peptide hormones in cancer. Discussion of possible cellular cascades involved in actions linking relaxin-like peptides and neoplasia include the role of relaxin-like peptides in tumor cell growth and differentiation; the effect of relaxin in stimulating the synthesis of the vasodilatory and tumor cell cytostatic and antiapoptotic molecule, nitric oxide; the potential ability of relaxin to upregulate vascular endothelial growth factor to promote angiogenesis and neovascularization and the concerted fine-tuned action of relaxin on the matrix metalloproteinases on the extracellular matrix to facilitate tumor cell attachment, migration and invasion. PMID- 14520687 TI - Inhibition of JNK signaling diminishes early but not late cellular stress-induced apoptosis. AB - The human leukemic T-cell line Jurkat was used to define the role of the cellular stress pathway with its key player kinase JNK in cancer therapy-induced apoptosis. JNK activity was inhibited by stable transfection with a dominant negative mutant of the upstream kinase JNKK/MKK4 or with the novel, potent and selective JNK1, -2 and -3 inhibitor SP600125. Inhibition of JNK activity delayed the onset of apoptosis induced by cisplatin, doxorubicin, gamma-irradiation and CD95-L but did not prevent apoptosis per se. Early events during apoptosis such as induction of CD95-L, activation of caspase-8 and exposure of phosphatidylserine on the cell surface were strongly inhibited. Also, at early time points of apoptosis, loss of the mitochondrial membrane potential and release of cytochrome c were markedly impaired. However, late signaling events during apoptosis such as cleavage of PARP and DNA fragmentation apoptosis were only marginally affected. These findings are in accordance with the activity of initiator and effector caspases. Whereas activity of the initiator caspase-8 was strongly inhibited early and late after induction, an inhibition of caspase-3 activity was only observed early after induction of apoptosis. We therefore suggest that cellular stress signaling contributes to the initiation of apoptosis, whereas it might be dispensable for the progression of apoptosis. Dysfunction of this pathway under pathological conditions might contribute to therapy resistance of cancer cells. PMID- 14520688 TI - Gene expression profiling of tumor xenografts: In vivo analysis of organ-specific metastasis. AB - Mammary carcinoma frequently metastasizes to specific organs, including the regional lymph nodes, the lung and bone marrow. The mechanisms that guide organ specific metastasis and the molecular players that are involved remain to be established. To gain insight into this problem, we used an orthotopic model of breast cancer in which the MDA-MB-435 cells are implanted into the mouse mammary fat pad. Sublines that preferentially metastasized to specific sites were isolated by excising metastatic tumors and growing explants of these tumors in culture. Cells lines that preferentially metastasize to the lymph node and thoracic cavity were obtained. The gene expression profiles of primary tumors from these sublines were then compared with cDNA arrays containing 5,800 known genes. In tumors that preferentially metastasize to the lymph node, several genes encoding adhesion and matrix proteins were upregulated. Genes encoding proteins involved in metabolism were downregulated. One of the upregulated genes in lymph homing tumors was CD73. Immunohistochemistry showed that the CD73 protein is also upregulated in primary tumors of this cell line and that its expression is elevated in the lymph node metastases. CD73 is a transmembrane protein that has previously been implicated in the homing of normal lymphocytes to the nodes. This raises the hypothesis that tumors preferentially metastasize to lymph nodes by using CD73 to mimic part of the lymphocyte homing process. PMID- 14520689 TI - Estradiol increases extracellular levels of vascular endothelial growth factor in vivo in murine mammary cancer. AB - Angiogenesis is essential for tumor growth and metastasis and an important prognostic factor in breast cancer. VEGF, a key factor for angiogenesis, has been correlated with tumor vessel density in breast cancer. Estrogen, another crucial factor in breast cancer, stimulates VEGF; and an ERE in the VEGF gene has been defined. VEGF is bioactive in the extracellular fluid, where it becomes available to endothelial cells. Whether E(2) affects VEGF levels in the extracellular fluid is not known. We show, using intratumoral microdialysis in vivo, that E(2) treatment increased tumor extracellular levels of VEGF in an estrogen-dependent breast cancer model. Moreover, extracellular levels of VEGF in the tumor showed a strong correlation with total tumor VEGF, contrary to plasma levels of VEGF. Ninety-three percent of measured VEGF in the extracellular fluid in the tumor was tumor-derived, while only 45% of VEGF in circulating plasma originated from the tumor. We conclude that E(2) increases extracellular VEGF and that microdialysis is a sensitive method for measurement of local VEGF production in vivo. Our results have potential application to the assessment of tumor characteristics in vivo in human tumors for individualized cancer therapy. PMID- 14520690 TI - Use of plasma MMP-2 and MMP-9 levels as a surrogate for tumour expression in colorectal cancer patients. AB - Matrix metalloproteinases, and notably the gelatinases MMP-2 and MMP-9, have important roles in tumour invasion, metastasis and angiogenesis. Our study investigates the distribution of MMP-2 and MMP-9 in colorectal cancer, the correlation with plasma levels, changes following surgical resection and whether plasma levels reflect clinical staging and disease course. MMP-2 and MMP-9 expression in 48 colorectal tumours and 13 adenomatous polyps was analysed by RT PCR, immunohistochemistry, and quantified by ELISA of tumour lysates. Concentrations of MMP-2 and MMP-9 in plasma samples from these patients and 36 other patients who underwent curative resections were measured by ELISA prior to and 6-12 months after surgery. MMP-2 expression was significantly increased in colorectal cancer tissues compared to matched normal colon as measured by ELISA. Active MMP-2 was localised by immunohistochemistry to regions where tumour cells invaded the muscularis with little staining in more superficial areas. Plasma MMP 2 levels were also significantly elevated in patients with colorectal cancer, with significant reductions following curative resections at all stages. Similarly, MMP-9 expression was significantly increased in colorectal cancer tissues, predominantly in the tumour stroma. Plasma levels of MMP-9 were significantly elevated at all stages in colorectal cancer patients and a significant reduction was seen following curative resections. With both MMP-2 and MMP-9, the strongest correlation with clinical staging in colorectal cancer was represented by the total plasma concentration of the enzymes, both falling to within the normal range following curative surgery. Plasma levels of these enzymes may therefore have potential as a noninvasive indicator of invasion or metastasis in colorectal cancer or as a marker of disease status during follow up. PMID- 14520691 TI - Expression of microsomal prostaglandin E synthase-1 in intestinal type gastric adenocarcinoma and in gastric cancer cell lines. AB - Gastrointestinal carcinomas synthesize elevated levels of prostaglandin E(2) (PGE(2)), which has been mechanistically linked to carcinogenesis. Recently, microsomal prostaglandin E synthase-1 (mPGES-1) was cloned, which seems to be inducible and linked to cyclooxygenase-2 (Cox-2) in the biosynthesis of PGE(2). We examined expression of mPGES-1 in intestinal type gastric adenocarcinomas and in gastric cancer cell lines. The transcript for mPGES-1 was elevated in 57% (4/7) of gastric carcinomas as detected by Northern blot analysis. Moderate to strong mPGES-1 immunoreactivity was observed in 56% (5/9) of the carcinomas as detected by immunohistochemistry. Furthermore, mPGES-1 mRNA, protein and microsomal PGES activity were detected in gastric adenocarcinoma cell lines that originated from intestinal type tumors (MKN-7 and MKN-28). In contrast to Cox-2, however, expression of mPGES-1 mRNA or protein were not induced by phorbol 12 myristate 13-acetate (PMA) or interleukin-1beta (IL-1beta) in any of the gastric cancer cell lines tested (MKN-1, -7, -28, -45 and -74). Two gastric cancer cell lines (MKN-45 and MKN-74) did not express mPGES-1 and lacked microsomal PGES activity, but were still able to synthesize PGE(2). Because all gastric cell lines expressed cPGES as detected by immunoblotting, it is possible that Cox-2 can interact with cPGES or with some other yet unidentified PGES in gastric cancer cells. Furthermore, our data show that regulatory mechanisms that drive expression of mPGES-1 and Cox-2 dissociate in gastric cancer cell lines. PMID- 14520692 TI - Suppression of peritoneal dissemination through protecting mesothelial cells from retraction by cancer cells. AB - In a previous study, we demonstrated that calponin h1 suppressed tumor growth of transformed cells and that the peritonitis carcinomatosa induced by mouse B16-F10 melanoma (F10) cells was more extensive in calponin h1-deficient (CN(-/-)) mice with fragility of mesothelial (MS) cells than in their calponin h1-wild (CN(+/+)) counterparts. In our study, we assessed the therapeutic effect of calponin h1 on peritoneal dissemination. F10 cells were overlaid on the cultured CN(+/+) or CN( /-) MS cells and the effect of calponin h1 on retraction of MS cells was evaluated. Then, an adenoviral vector with the calponin h1 gene (AdGFP-CN) inserted was constructed and was applied to CN(-/-) MS cells or CN(-/-) mouse peritoneum to investigate its suppressive effect on the peritoneal dissemination caused by F10 cells. Greater retraction and invasion of F10 cells were observed in CN(-/-) MS than in CN(+/+) cells in vitro, while down-regulation of calponin h1 was observed in CN(+/+) MS cells prior to the invasion of F10 cells. Infecting CN(-/-) MS cells with AdGFP-CN prevented their retraction and the invasion of F10 cells. Peritoneal dissemination was prominently suppressed in AdGFP-CN-infected CN(-/-) mice, and the survival of those mice was significantly prolonged. Thus, calponin h1 functioned to protect host MS cells from the invasion of F10 cells. PMID- 14520693 TI - Increased expression of the mannose 6-phosphate/insulin-like growth factor-II receptor in breast cancer cells alters tumorigenic properties in vitro and in vivo. AB - The mannose 6-phosphate/insulin-like growth factor-II receptor (M6P/IGF-IIR) is thought to act as a suppressor of tumor growth by binding the mitogenic peptide IGF-II and modulating its extracellular levels via degradation. This receptor has been found to be absent or nonfunctional in a high proportion of breast tumors as a result of LOH and mutation of the gene. In our study, we have examined the effect of increasing expression of M6P/IGF-IIR on breast cancer cell tumorigenicity. MDA-MB-231 breast cancer cells stably transfected with M6P/IGF IIR cDNA exhibited not only a greatly reduced ability to form tumors but also a markedly reduced growth rate in nude mice. In vitro, increased M6P/IGF-IIR expression resulted in 2-fold reduced uptake of IGF-II and was associated with reduced cellular invasiness and motility. Cells with increased M6P/IGF-IIR expression exhibited reduced phosphorylation of IGF-I receptor and p44/42 MAPK compared to vector transfectants, or wild-type MDA-MB-231 cells. These results therefore suggest that M6P/IGF-IIR levels can modulate breast cancer cell tumorigenicity by a mechanism that may involve altered IGF-I receptor signaling. PMID- 14520694 TI - MSH6 germline mutations are rare in colorectal cancer families. AB - Germline mutations in MSH6 can cause HNPCC, which is associated with a tumor phenotype featuring MSI. However, tumors arising in persons with disease-causing mutations of MSH6 may or may not exhibit MSI. We used D-HPLC to screen for germline mutations in the promoter region, the coding region and the 3'-UTR of MSH6. Eighty-four families, enrolled on the basis of Amsterdam I and II criteria (HNPCC families) and less stringent criteria (HNPCC-like families), were tested for MMR gene mutations; 27 families had a disease-causing mutation in MLH1 or MSH2, and the remaining 57 families were tested for mutations in MSH6. Two protein-truncating mutations were identified in each of 2 families fulfilling the Amsterdam I criteria, being present in persons affected with early-onset colorectal cancers exhibiting MSI. Immunohistochemical analysis showed that expression of both MSH2 and MSH6 proteins was lost in the cancer cells of the 2 mutation carriers but only MSH6 protein expression was lost in 2 adenomatous polyps. A third possibly disease-causing mutation was found in a person affected with a tumor that did not exhibit MSI. In addition, we found 4 new polymorphisms and determined that neither of the 2 studied by association analysis conferred susceptibility to colorectal or endometrial cancer. Altogether, our results indicate that disease-causing germline mutations of MSH6 are rare in HNPCC and HNPCC-like families. PMID- 14520695 TI - Microsatellite instability in hereditary and sporadic breast cancers. AB - Sporadic cancers and familial breast cancers are characterized by an increase in genetic instability. Little is known about whether mismatch repair defects accompany this genetic instability. We investigated invasive and/or in situ breast cancers from 30 women with deleterious BRCA1/2 mutations and unclassified variant BRCA1/2 alterations. Forty cases of sporadic breast cancers were also investigated, including 7 medullary carcinomas. Malignant and benign lesions were examined from all cases to better understand tumor progression. Automated immunohistochemistry, with antibodies directed against hMLH1 and hMSH2, was used to screen cases for possible mismatch repair defects. When loss of expression was noted, DNA ploidy was performed by cytomorphometry. DNA, after laser microdissection, was extracted from a majority of familial cases and their corresponding controls, and microsatellite instability analysis was performed. None of the familial or sporadic cases had loss of hMSH2 expression. All but one lesion, a DCIS arising in a deleterious BRCA2 mutation carrier, had loss of hMLH1 expression and a tetraploid profile by image cytomorphometry. There was no MSI in any explored lesions (n = 34), as determined by molecular analysis, including the DCIS with loss of hMLH1 expression. We conclude that DNA mismatch repair defects involving hMLH1 and hMSH2 underexpression are extremely rare events in sporadic and familial breast cancer. Mismatch repair gene mutations may be secondary random events in breast cancer progression. PMID- 14520696 TI - Breast cancer in young women (< or = 35 years): Genomic aberrations detected by comparative genomic hybridization. AB - Sporadic breast cancer in young women is different from the one in older patients regarding pathological features and aggressiveness of the tumors, but the spectrum of genetic alterations are largely unknown. We used comparative genomic hybridization (CGH) to analyze DNA copy number changes in 88 tumor samples from women 0.81) to ID(50) values for berberine and C. rhizoma, respectively. Among these 33 genes, the levels of expression of 12 were correlated with the ID(50) values of both agents, suggesting that these genes are associated with tumor-killing activity of berberine in C. rhizoma. Expression of the remaining 21 genes was correlated with the ID(50) value of either purified berberine or C. rhizoma. Thus, we identified common and distinct genes responsible for anti-proliferative activities of purified berberine and C. rhizoma. This strategy may improve our understanding of the actions of herbs with antitumor activities. PMID- 14520709 TI - EGF +61 gene polymorphism and susceptibility to and prognostic markers in cutaneous malignant melanoma. AB - CMM is the most serious cutaneous malignancy and is increasing in frequency among most Caucasian populations, where the most important risk factor is exposure to UV light. Relatively little is known of the genetic factors that mediate susceptibility to and prognosis in sporadic CMM, although a number of genes have been implicated. A striking association between EGF polymorphism and Breslow thickness of invasive CMM has been reported. We have sought confirmation of this finding in an independent study of 159 patients and 310 controls using TaqMan fluorescence-based genotyping for EGF +61. In our study group, there were no significant differences in EGF genotype frequencies between patients and controls nor was EGF genotype associated with tumour growth phase, stage or mitotic count. However, correlation between EGF genotype and Breslow thickness showed a modestly significant increase in frequency of the EGF (G/G) genotype among tumours >3.5 mm thick (30.0% vs. 9.8%, p = 0.03). In summary, in our group, the EGF +61 polymorphism was not a risk factor for CMM susceptibility, but this polymorphism may play a role in disease progression. PMID- 14520710 TI - Increase of AKT/PKB expression correlates with gleason pattern in human prostate cancer. AB - AKT/PKB is a central signaling molecule related to stimulation of cell proliferation and inhibition of apoptosis. Perturbations of AKT expression and function play an important role in tumor development and progression. We wanted to determine (a) whether AKT is overexpressed in human prostatic tumors, (b) whether AKT expression is correlated with tumor grade, and (c) whether AKT expression correlates with clinicopathological parameters. AKT expression was investigated by immunohistochemistry in sections from 56 paraffin-embedded prostate specimens displaying benign prostatic tissue (BPT), prostatic intraepithelial neoplasia (PIN), and primary tumors graded 2-5 according to Gleason. The staining intensity for AKT was significantly more pronounced in tumors compared to BPT, with PIN ranging between BPT and carcinomas. Similarly, the fraction of AKT-positive cells was higher in tumors than in BPT. A score of AKT expression (calculated as product from intensity and fraction of positive cells) ranging from 0-6 was also significantly higher in tumors than in BPT. Furthermore, the intensity of AKT expression in tumors showed a positive correlation with high preoperative serum levels of prostate specific antigen (PSA >/= 10 ng/ml, p = 0.0325). These data show that AKT is upregulated in prostate cancer and that expression is correlated with tumor progression. PMID- 14520711 TI - Chlamydia pneumoniae infection and the risk of female early-onset lung cancer. PMID- 14520712 TI - Influence of tumor location on breast cancer prognosis. PMID- 14520713 TI - Re: Mesothelioma is a killer of urban men in Sweden by Kari Hemminki and Xinjun Li. PMID- 14520715 TI - Into the 21rst century. PMID- 14520716 TI - Developmentally appropriate healthcare for young people with chronic illness: questions of philosophy, policy, and practice. PMID- 14520717 TI - Breastfeeding and asthma: appraising the controversy--a rebuttal. PMID- 14520718 TI - New therapies for asthma: where next? PMID- 14520719 TI - Perspectives of patients with cystic fibrosis on preventive counseling and transition to adult care. AB - The purpose of this study was to investigate how adolescents and adults with cystic fibrosis (CF) view preventive counseling and their transition to adult centered care within a children's hospital. Thirty-two patients >/=16 years old diagnosed with CF were recruited from a pediatric tertiary care setting. During face-to-face interviews, patients were asked 27 structured questions and completed a 30-item self-administered questionnaire on preventive counseling by healthcare providers and on transition issues. The median age of patients was 25.5 years (range, 16-43 years); 69% of patients identified a pulmonologist as their "main doctor," even though 78% had a primary care provider. Participants felt that 13-16 years of age was the best time for them to begin spending time alone with their main doctor. Less than half of the participants recalled receiving preventive counseling during the previous 12 months, and more patients wanted to discuss issues than actually did. Qualitative data emphasized the importance of independence in making decisions in healthcare and establishing relationships with providers, and many patients did not desire to transfer care to an adult hospital. Participants identified adult-focused services such as inpatient rooms, discussion groups, work options, and social service support that would enhance care. In conclusion, the majority of adolescent/young adult patients with CF receiving care in a pediatric institution reported satisfaction with their healthcare. However, patients identified preventive issues that they desired to be more regularly addressed, starting in early adolescence, and changes in the delivery of services to enhance transition to adult-oriented care. This study underscored the understanding of the integration of transition planning into the facilitation of healthcare decision-making by the adolescent in issues of self-care, sexuality, education, and finances. Future initiatives to enhance the care of patients with CF should provide training of pulmonologists in preventive care and increased attention to helping patients utilize appropriate primary-care services during the adult years. In addition, prospective studies are needed to compare outcomes of CF patients who have transitioned and transferred to adult hospitals and those transitioning to adult-oriented services in a pediatric institution. PMID- 14520720 TI - Pneumonia due to Chlamydia pneumoniae in children: epidemiology, diagnosis, and treatment. AB - The term "atypical" pneumonia has been used to differentiate infections caused by Chlamydia pneumoniae, Mycoplasma pneumoniae, Legionella, and other related organisms from pneumonia caused by classic bacteria, the prototype being Streptococcus pneumoniae. However, recent studies demonstrated that the clinical presentation of pneumonia due to atypical pathogens cannot readily be differentiated from those caused by "typical" bacteria. This is further complicated by the observation that coinfections with atypical pathogens and other bacteria are frequent. Nonetheless, the term "atypical" can be useful, as these organisms share a number of characteristics that separate them from "typical" bacteria. They are either obligate or facultative intracellular parasites that cannot be isolated using routine microbiologic methods. The most commonly used method of diagnosis of these infections is serology, which has significant limitations. Although C. pneumoniae is now recognized worldwide as a common cause of respiratory infections in adults and children, major gaps remain in our knowledge of the biology of this organism and how it causes disease, in major part due to the lack of readily available, standardized diagnostic methods. PMID- 14520721 TI - Effect of long-acting beta2 agonists on exacerbation rates of asthma in children. AB - The purpose of this analysis was to examine the effect of long-acting beta(2) adrenoceptor agonists (LABAs) on the asthma exacerbation rate in pediatric patients. Randomized controlled trials (RCT) that included the use of LABAs to treat symptoms of pediatric asthma in children on inhaled corticosteroids, that reported asthma exacerbation rates, and that were published as full papers in peer-reviewed journals were retrieved from a search of the medical literature. Eight studies were identified that fulfilled these criteria. An exacerbation was defined as deterioration in a patient's asthma requiring a change in prescribed medication or not defined but reported as an asthma exacerbation or an asthma related hospitalization. Analysis of data from the eight studies revealed no apparent protection from an asthma exacerbation among children on a LABA compared to patients on comparator treatment. The relative risk of an asthma exacerbation for LABA compared to placebo or short-acting beta(2)-adrenoceptor agonist (SABA) ranged from 0.95-1.86. The relative risk of hospitalization for asthma in patients treated with LABAs with regular maintenance with ICS ranged from 3.3 21.6 in the three studies that reported asthma-related hospitalizations. The lack of evidence for the control of asthma exacerbations in children regularly using a LABA should bring into question its general use as add-on therapy. Studies should be designed to directly explore the implications of these observations in pediatric patients. PMID- 14520722 TI - Does vitamin D administered to children with asthma treated with inhaled glucocorticoids affect short-term growth or bone turnover? AB - Our objective was to assess whether administration of 25-OH-vitamin D to children with asthma treated with inhaled dry-powder budesonide 400 microg daily affects short-term growth or markers of bone turnover. We utilized a randomized, double blind, two-period crossover trial with run-in and washout periods of 2 weeks and treatment periods of 4 weeks duration. The setting was an Outpatient clinic in a secondary referral center. Subjects included 14 boys and 3 girls with a mean age of 11.7 (range, 6.1-14.4) years. Interventions included 15 microg (600 IU) 25-OH vitamin D (cholecalciferol) in one tablet ABCDin(R) once daily in the morning. Primary outcome measures were: lower leg growth rate, serum osteocalcin, and serum markers of type I collagen turnover, i.e., the amino terminal propeptide of type I procollagen (PINP), the carboxy terminal propeptide of type I procollagen (PICP) (formation markers), and the carboxy terminal pyridinoline cross-linked telopeptide of type I collagen (ICTP) (degradation markers). Secondary outcome measures were parameters of asthma control and serum 25-OH-vitamin D. Lower leg growth rate was 0.22 mm/week during vitamin D and 0.25 mm/week during placebo treatment (NS). Osteocalcin was 59.9 and 57.8 microg/l during vitamin D and placebo treatment, respectively, PINP 574 and 565 microg/l, PICP 381 and 382 microg/l, and ICTP 11.5 and 11.1 microg/l, respectively (NS). Serum 25-OH-vitamin D was 76.3 nmol/l and 48.2 nmol/l, respectively (P < 0.001). There were no statistically significant differences in measures of pulmonary function. In conclusion, administration of 25-OH-vitamin D does not affect short-term growth or markers of bone turnover in children with asthma treated with inhaled dry powder budesonide 400 microg daily. PMID- 14520723 TI - Pseudomonas aeruginosa and cystic fibrosis: correlation between exoenzyme production and patient's clinical state. AB - In this study, we investigated the correlation between the production by Pseudomonas aeruginosa isolates of four exoenzymes (protease, elastase, neuraminidase, and phospholipase C (PLC)) and the clinical state of cystic fibrosis (CF) patients. We studied 212 P. aeruginosa isolates from 22 CF patients chronically infected with this bacterium. Patients were classified into three clinical groups according to a modified Shwachman-Kulczycki-Khaw (SKK) scoring system. The production of enzymes by isolates from patients in the three populations was analyzed and compared using four statistical tests: chi-square, Mann-Whitney U, principal component analysis, and discriminant analysis. Isolates from patients with excellent or good clinical status (group I, SKK score >/=71) had higher elastase and neuraminidase activities than isolates from the other patients. In contrast, PLC activity, a common characteristic of CF isolates, was higher in isolates from patients with poor or weak clinical status (group III, SKK score FeIII-NO2 transformation in the presence of dioxygen. PMID- 14520755 TI - Enantioselective conjugate radical addition to beta-acyloxy acrylate acceptors: an approach to acetate aldol-type products. PMID- 14520757 TI - Supramolecular adducts between poly-L-arginine and [TmIIIdotp]: a route to sensitivity-enhanced magnetic resonance imaging-chemical exchange saturation transfer agents. PMID- 14520756 TI - Highly active catalysts in alkene metathesis: first observed transformation of allenylidene into indenylidene via alkenylcarbyne--ruthenium species. PMID- 14520758 TI - [tBuLi.(-)-sparteine]: molecular structure of the first monomeric butyllithium compound. PMID- 14520759 TI - Stepwise solid-phase synthesis of di- and trinuclear metal complexes. PMID- 14520760 TI - Bond-stretch isomerism: trapped isomeric structures of hexacoordinate copper(II) bispidine chromophores along a Jahn-Teller active vibrational coordinate. PMID- 14520761 TI - Aqueous chemistry of titanium(II) species. PMID- 14520762 TI - Knotaxanes--rotaxanes with knots as stoppers. PMID- 14520763 TI - Conformative coupling of two conformational molecular switches. PMID- 14520765 TI - Sleep bruxism: issues to chew on. PMID- 14520766 TI - Topical review: sleep bruxism and the role of peripheral sensory influences. AB - Sleep bruxism (SB) is an unusual orofacial movement described as a parafunction in dentistry and as a parasomnia in sleep medicine. Since several peripheral influences could be involved in sleep-wake regulation and the genesis of rhythmic jaw movements, the authors have reviewed the relevant literature to facilitate understanding of mechanisms possibly involved in SB genesis. Various animal and human studies indicate that during either wakefulness or anesthesia, orofacial sensory inputs (e.g., from periodontium, mucosa, and muscle) could influence jaw muscle activity. However, the role of these sensory inputs in jaw motor activity during sleep is unclear. Interestingly, during sleep, the jaw is usually open due to motor suppression; tooth contact most likely occurs in association with sleep arousal. Recent physiologic evidence supports an association between sleep arousal and SB; a sequential change from autonomic (cardiac) and brain cortical activities precede SB-related jaw motor activity. This suggests that the central and/or autonomic nervous systems, rather than peripheral sensory factors, have a dominant role in SB genesis. However, some peripheral sensory factors may exert an influence on SB through their interaction with sleep-wake mechanisms. The intent of this review is to integrate various physiologic concepts in order to better understand the mechanisms underlying the genesis of SB. PMID- 14520767 TI - Spread and referral of experimental pain in different jaw muscles. AB - AIMS: To test the hypothesis that there would be no differences in perceived pain intensity and spread and referral of pain evoked by injection of a similar amount of hypertonic saline into 6 different jaw-muscle sites in healthy female subjects. METHODS: A total of 15 healthy women participated in 3 experimental sessions separated by 1 week. In a randomized sequence, the deep layers of the masseter, superficial layers of the masseter, anterior temporalis, lateral pterygoid, medial pterygoid, and anterior digastric muscles were injected with 5.8% hypertonic saline (0.2 mL). The subjects rated the perceived intensity of pain on an electronic 0- to 10-cm visual analog scale (VAS). The distribution of pain was drawn by the subjects on anatomical maps of the face, and a Danish version of the McGill Pain Questionnaire (MPQ) was filled out. RESULTS: All injections were associated with moderate to strong pain intensity (mean peak value: 5.6 to 6.4 cm) with no significant differences between muscle sites (analysis of variance [ANOVA]: P = .520). Pain rating indices derived from the MPQ did not suggest significant differences between muscle sites (ANOVA: P = .898). However, the area of perceived pain differed significantly between muscle sites (ANOVA: P = .038) with the greatest area following the injection into the anterior temporalis muscle (Tukey: P < .05). On direct inspection, the pain maps appeared quite similar, but a new analysis technique based on a center-of-gravity method revealed significantly different coordinates and length of vectors (ANOVA: P < .001) with longer vectors associated with the pain areas in the anterior temporalis muscle compared with the other muscle sites (Tukey: P < .05). All muscles were frequently associated with referral of pain to intraoral structures (40% to 87%), but only pain in the anterior digastric muscle was referred to the tip of the tongue (53%). CONCLUSION: The data suggest no major differences in pain sensitivity between the examined jaw-muscle sites, but pain in the anterior temporalis muscle spreads to a larger area independent of pain intensity. There are subtle but detectable differences in the location and referral of pain patterns between jaw muscles. This will be helpful in the differential diagnosis of myofascial temporomandibular disorder pain. PMID- 14520768 TI - Use of complementary and alternative medicine for temporomandibular disorders. AB - AIMS: Despite many reports about complementary and alternative medicine (CAM) use in the general population, little information exists about specific CAM therapies used for particular health conditions. This study examines the use of CAM therapies among patients with temporomandibular disorders (TMD). METHODS: We surveyed 192 patients with documented TMD as part of a larger project on the effectiveness of various CAM modalities for TMD patients. The survey asked about use of and attitudes toward specific CAM therapies for treating TMD and other patient-identified health conditions. The survey also measured physical health, health behavior, and psychosocial functioning. RESULTS: Nearly two thirds of the respondents (62.5%; n = 120) reported using CAM therapies for TMD or a related condition. Of all the therapies reported, massage was rated as the most frequent and among the most satisfactory and helpful. In general, respondents who used CAM for their TMD reported being most satisfied with the "hands on" CAM therapies (massage, acupuncture, and chiropractic care). The vast majority of respondents reported using CAM approaches for TMD simultaneously with conventional care (95.6%; 66 of 69). Those using CAM for TMD tended to be older, had a history of multiple medical problems, and reported more positive psychologic functioning. Respondents who most often reported CAM treatment as "very helpful" for their TMD were likely to be healthier (i.e., reporting higher levels of exercise and fewer sleep disturbances). CONCLUSION: Given the frequent use of CAM treatments by our respondents, allopathic providers should inquire about the adjunctive use of CAM among their TMD patients. PMID- 14520769 TI - Analgesic efficacy of low-dose diclofenac versus paracetamol and placebo in postoperative dental pain. AB - AIMS: To compare the efficacy and safety of diclofenac-K (12.5 mg) vs paracetamol (500 mg) and placebo given in a flexible dosage regimen to treat pain resulting from extraction of impacted third molar teeth. METHODS: This was a 2-day, double blind, double-dummy, randomized, parallel-group, placebo-controlled study of diclofenac-K (12.5 mg) tablets vs paracetamol (500 mg) tablets and placebo in patients with moderate or severe pain within 8 hours of extraction of impacted third molars. RESULTS: After the first 2-tablet dose, patients took on average 2.5 additional tablets of diclofenac-K or 2.4 tablets of paracetamol, almost all as 1-tablet doses. Most placebo patients discontinued by taking rescue medication (ibuprofen 200 mg) on the first day. Pain relief after the initial dose of diclofenac-K (2 x 12.5 mg) was superior to placebo (P < .01 for all efficacy outcomes) and comparable to paracetamol (2 x 500 mg). About 30% of patients in each active treatment group took rescue medication during the study, compared to 78% on placebo. About 70% in each active treatment group considered the overall pain relief to be "some," "a lot," or "complete" compared to only 15% on placebo. The incidence of adverse events in each active treatment group was low and comparable between the treatments. CONCLUSION: An initial double-dose of diclofenac-K (2 x 12.5 mg) or paracetamol (2 x 500 mg) adequately relieved the most intense postoperative pain, and the flexible multiple dose regimen (1 or 2 tablets) maintained adequate pain relief thereafter. Most patients needed only 1 tablet doses following the initial 2-tablet dose. PMID- 14520771 TI - The effect of exogenous glucosamine hydrochloride on the proteoglycan concentration of the articular disc of the rabbit temporomandibular joint. AB - AIMS: To test the effect of glucosamine hydrochloride (glucosamine-HCl) on the proteoglycan (PG) concentration of the articular disc of non-arthritic temporomandibular joints (TMJs) in rabbits. METHODS: Twenty-four of 48 New Zealand white 10- to 12-week-old male rabbits (2.2 kg average) were injected with the irritant chymopapain in the knee joint. Both groups of 24 rabbits were divided into 3 groups of 8 animals. The rabbits were fed a control diet or a diet supplemented with glucosamine-HCl incorporated at a level to provide 20 mg/kg (approximating the recommended dose 500 mg/tid/70 kg man) or 100 mg/kg, for 8 weeks. Sulfated glycosaminoglycans (GAGs) were assayed in protease K digests of the TMJ articular disc by dimethylmethylene blue method to quantify PG concentration. The groups were compared with 2-way analysis of variance. RESULTS: Glucosamine-HCl did not cause a significant change in the PG concentration of the TMJ articular disc (P > .8). There was also no detectable effect of chymopapain injection to the knee joint on the TMJ (P > .07) and no interaction between glucosamine-HCl treatment and chymopapain injection (P > .3). CONCLUSION: Glucosamine-HCl has no effect on PG concentration of the articular disc of non arthritic TMJ in rabbits. PMID- 14520770 TI - Capsaicin receptor VR1 and ATP purinoceptor P2X3 in painful and nonpainful human tooth pulp. AB - AIMS: To investigate the levels of the capsaicin or vanilloid receptor-1 (VR1) and the ATP-gated purinoceptor P2X3 in painful and nonpainful human tooth pulps. METHODS: Immunohistochemistry with specific antibodies and image analysis was used to quantify VR1- and P2X3-positive nerve fibers in painful (n = 13) and nonpainful (n = 33) human tooth pulps, and VR1 immunoreactivity was compared with immunoreactivity for the structural neuronal marker peripherin. RESULTS: Strong VR1-like immunoreactivity was documented for the first time in dental pulp neurons. Weaker P2X3-like immunoreactivity was also detected in fewer nerve fibers. The ratio of VR1 to peripherin immunoreactivity was not significantly different between nonpainful and painful tissues (mean +/- SE% area of VR1 to peripherin; nonpainful 53.4 +/- 4.7%, n = 33; pulpitis 35.1 +/- 7.1%, n = 13; P = .07). CONCLUSION: The presence of VR1 and P2X3 in fibers of human tooth pulp suggest that they may play a role in perception of dental pain, but further studies, including quantitation of their ligands, are necessary to elucidate any role they may play in pathophysiologic states. PMID- 14520772 TI - Mandibular condyle osteochondroma. Review of the literature and report of a misdiagnosed case. AB - Temporomandibular disorders can usually be diagnosed on the basis of a thorough history and a comprehensive examination of the patient. Additional diagnostic tests, such as imaging of the temporomandibular joint (TMJ) area, are mandatory and must be flawless in case of atypical findings. The aim of this report is to illustrate pitfalls in clinical reasoning and in imaging procedures in the diagnosis of temporomandibular pain and dysfunction. A case report of a patient with osteocartilaginous exostosis of the mandibular condyle, which was erroneously diagnosed and treated as an internal derangement of the TMJ for half a year, is presented. PMID- 14520773 TI - A comparison of a waterpik dual-motor powered toothbrush and a manual toothbrush in affecting interproximal bleeding reduction and dental biofilm accumulation. AB - OBJECTIVE: This single-blind, 21-day clinical study was designed to compare the effectiveness of a dual-motor powered toothbrush (SynchroSonic) and a manual toothbrush in affecting interproximal bleeding reduction and dental biofilm accumulation in a cohort of 70 subjects. METHODOLOGY: After baseline evaluations of soft tissue trauma, dental biofilm and interproximal bleeding, subjects were randomly assigned to one of the two experimental groups, given toothbrushing instructions and had their teeth cleaned. At 21 days, the clinical parameters were again evaluated. RESULTS: In the powered toothbrush group, there were 37 subjects at baseline and 36 subjects at follow-up. In the manual toothbrush group, there were 34 subjects at baseline and 33 subjects at follow-up. In the powered toothbrush group, the mean biofilm index fell from 3.46 to 3.12; a 9.7% reduction. In the manual toothbrush group, the mean biofilm index fell from 3.51 to 3.30; a 5.4% reduction. The powered toothbrush group showed significantly greater biofilm reduction (9.7% vs. 5.4%, p = 0.043). In the powered toothbrush group, the mean gingival bleeding index fell from 0.66 to 0.44; a 32% reduction. In the manual toothbrush group, the mean gingival bleeding index fell from 0.66 to 0.53; a 20% reduction. The powered toothbrush group showed significantly greater gingival bleeding reduction (32% vs. 20%, p = 0.02). CONCLUSION: The results demonstrated that both the manual toothbrush and the SynchroSonic powered toothbrush were safe, with no evidence of clinically significant hard or soft tissue abrasion observed. Additionally, the powered toothbrush was more effective in reducing biofilm and gingival bleeding than the manual toothbrush. PMID- 14520774 TI - Dental caries in school-age children residing in five Guatemalan communities. AB - OBJECTIVE: The caries epidemiology within the country of Guatemala is poorly understood. This work reports the cross-sectional prevalence of caries in the permanent teeth of children between the ages of 6 and 12 years in five different communities within Guatemala, and relates the caries experience to the fluoride levels in the communal drinking water in each of these cities. METHODOLOGY: These cities were selected because they represent geographically distinct regions of Guatemala, where there is no fluoridation of public drinking water. A total of 1,145 children were enrolled and examined in this study, with 227, 230, 231, 228 and 229 enrolled at Coban, Solola, Guatemala City, Chiquimula, and Estanzuela, respectively. Caries examinations were performed by a calibrated examiner using a visual tactile examination method performed with the aid of an artificial light, mouth mirror, compressed air, and a dental explorer employing a modified Radike criteria. Fluoride concentrations from drinking water, collected from the primary drinking water spigot at each elementary school in each city, were determined using ion chromatography. RESULTS: The cross-sectional mean DMFS in children 6-8 years old was 4.61, 4.34, 3.75, 3.52 and 2.71 for Coban, Solola, Guatemala City, Chiquimula, and Estanzuela, respectively. The cross-sectional mean DMFS in children 9-12 years old was 10.96, 10.09, 8.67, 8.71 and 6.02 for Coban, Solola, Guatemala City, Chiquimula, and Estanzuela, respectively. The prevalence of caries in children between the ages of 9 and 12 years was greater than 90% in all five cities. The natural fluoride concentration in water was 0.05 ppm, 0.23 ppm, 0.14 ppm, 0.50 ppm, and 0.60 ppm for Coban, Solola, Guatemala City, Chiquimula, and Estanzuela, respectively. CONCLUSION: Collectively, these data demonstrate that the high prevalence of caries in Guatemala appears to be directly correlated to levels of fluoride in the community drinking water. PMID- 14520775 TI - Clinical response of two brush-applied peroxide whitening systems. AB - OBJECTIVE: A randomized, examiner-blind parallel design study was conducted to evaluate the clinical efficacy and tolerability of two brush-applied, peroxide based tooth whiteners. METHODOLOGY: A total of 38 subjects were randomized to Colgate Simply White, an 18% carbamide peroxide paint-on liquid in an applicator bottle, or Crest Night Effects, a 19% sodium percarbonate system in unit dose sachets that dries to form an adherent film. Treatment was for 14 days. The 18% carbamide peroxide gel was applied twice daily, while the 19% sodium percarbonate film was applied for overnight use. Subjects were evaluated at baseline, and again after 7 and 14 days of treatment using digital images to assess tooth color, and oral examinations to establish safety. Whitening efficacy was determined by evaluating the reduction in tooth yellowness (delta b*), increase in tooth brightness (delta L*), reduction in tooth redness (delta a*) and overall tooth color change relative to pure white (delta W*). RESULTS: After 14 days, the mean change in yellowness (delta b*) was -1.43 +/- 0.20 for the 19% sodium percarbonate film, and -0.44 +/- 0.06 for the 18% carbamide peroxide gel. Mean change in brightness (delta L*) was 1.13 +/- 0.13 and 0.55 +/- 0.13 in the film and gel groups, respectively. Between-group comparisons showed highly significant (p < or = 0.003) whitening for the 19% sodium percarbonate film compared to the 18% carbamide peroxide gel for all 14-day color measurements (delta b*, delta L*, delta a* or delta W). Both products were well-tolerated, with no subjects discontinuing treatment early due to a causal adverse event. CONCLUSION: In a direct clinical comparison of two brush-applied products over 14 days, Crest Night Effects provided significant and meaningful improvement in tooth color, representing more than double the whitening measured for the Colgate Simply White. PMID- 14520776 TI - Comparative clinical investigation of the tooth whitening efficacy of two tooth whitening gels. AB - OBJECTIVE: The objective of this randomized, double-blind, parallel-group clinical study was to compare the tooth whitening efficacy of two tooth whitening gel products--Colgate Simply White Night Clear Whitening Gel containing either 25% carbamide peroxide, or 8.7% hydrogen peroxide--when used once daily at night. METHODOLOGY: Following a baseline tooth shade evaluation using the Vita Shade Guide, qualifying adult male and female subjects from the Central New Jersey area were randomized into two treatment groups, which were balanced for baseline Vita Shade Guide scores, gender and age. The treatment groups were: 1) a tooth whitening gel containing 25% carbamide peroxide; or 2) a tooth whitening gel containing 8.7% hydrogen peroxide. All subjects were given a commercially available, non-whitening dentifrice and a soft-bristled toothbrush at the beginning of the study. In addition, they were also given one of the two tooth whitening gel products. All subjects were instructed to brush their teeth for one minute twice daily (morning and evening) with the non-whitening toothpaste. The subjects were further instructed to apply their assigned tooth whitening gel product once daily at night, per the instructions provided. Evaluations of tooth shade for each subject were repeated after two weeks, and again after three weeks of product use. RESULTS: Fifty-nine (59) subjects complied with the protocol and completed the three-week study. At both the two-week and three-week examinations, all subjects exhibited statistically significant (p < 0.05) tooth shade lightening relative to baseline tooth shade. Further, there was no statistically significant difference in tooth shade lightening between the two tooth whitening gel products. CONCLUSION: The results of this clinical study indicate that after once daily use at night for two or three weeks, a tooth whitening gel containing 25% carbamide peroxide and a tooth whitening gel containing 8.7% hydrogen peroxide both provided statistically significant tooth shade lightening relative to baseline tooth shade. The results also showed that there was no statistical difference in tooth whitening efficacy between the two tooth whitening gel products. PMID- 14520777 TI - Plaque removal and oral health promotion potential for the elmex interX medium toothbrush: clinical efficacy and safety evaluation. AB - OBJECTIVE: This study was designed to assess a newly developed toothbrush for efficacy in plaque removal, reduction of gingivitis and soft tissue bleeding, and retardation of stain formation over 30 days. METHODOLOGY: Healthy adult subjects (N = 112) were admitted to the study, and randomly given a test toothbrush, (the elmex interX medium) or the control toothbrush, an ADA-Accepted flat-bristle profile toothbrush. Additionally, the subjects were randomly assigned to be given oral hygiene instruction, or no formal oral hygiene instruction. Examinations and assessments were performed at baseline, 15 and 30 days, and included gingivitis levels, stain levels, bleeding on probing, and plaque levels before and after brushing. RESULTS: No significant statistical differences were found between the two toothbrushes on gingivitis, nor on the stain index; however, over the course of the study the gingivitis decreased significantly (p = 0.012) for the test toothbrush group. For the bleeding index, a significant statistical difference (p = 0.027) was found at 30 days favoring the test toothbrush. There was also a significant decrease for the test toothbrush on bleeding over time (p = 0.004). Following overnight plaque formation, the before-brushing plaque levels were significantly different in favor of the test toothbrush at 15 days (p = 0.002), and again at 30 days (p = 0.001). Immediately after brushing, there was a significant statistical difference (p = 0.015) favoring the test toothbrush at 30 days. CONCLUSION: These results clearly indicate improved plaque removal and reduction of bleeding and gingivitis for the new elmex interX medium toothbrush compared to the ADA-Accepted control toothbrush. PMID- 14520779 TI - A settlement in Boston. PMID- 14520778 TI - Verification of caries inhibition by a tartar control toothpaste. AB - OBJECTIVE: The anticaries properties of a silica-based, sodium fluoride (NaF) toothpaste containing tetrapotassium pyrophosphate (PPi) with anti-tartar properties (Aquafresh All Tartar Control) were assessed in specific pathogen-free Osborne-Mendel rats. METHODOLOGY: The dentifrice was compared to a silica-based, fluoride-free control toothpaste that contained PPi but no NaF, termed a "Placebo" (negative control), and a NaF-containing silica-based USP reference standard toothpaste which was used as a positive fluoride control toothpaste. Sixty weanling rats were randomly distributed into groups of 20; all were inoculated with S. mutans 10449S, ate cariogenic diet 2000, and drank demineralized water, ad libitum. Each toothpaste, contained in coded tubes, was applied to the dentitions of rats' teeth for one minute, twice daily on weekdays and once daily on weekends and holidays. There was no difference in weight gains or in recoveries of the S. mutans inoculant from tooth swabs among the groups at either the 24- or 38-day post-inoculation recovery date. After euthanasia at 42 days and defleshing, heads were randomly coded and scored blindly for carious lesions according to Keyes/Larson methods. Only after scoring was completed were the random codes broken to enable statistical analyses. Only after statistical analyses were completed and the data reported to sponsor were the identities of the test items revealed. RESULTS: Both the NaF/PPi-containing and the NaF containing USP standard toothpaste groups had lower total enamel caries scores (29.8 to 30.6%, respectively) than the group treated with the fluoride-free PPi containing control. Similar-dimensioned differences were evident both at smooth surface and sulcal enamel sites and in dentinal sites. All were statistically significant at p < 0.001. There were no statistically significant differences at any tooth surface category site between the two fluoride-containing toothpastes' effects. CONCLUSION: It is thus apparent that Aquafresh All Tartar Control, an anti-tartar NaF toothpaste, retains the anticaries benefit of a USP reference standard NaF toothpaste in vivo. PMID- 14520780 TI - The fitness of the republic. PMID- 14520781 TI - Casting a wide net. PMID- 14520782 TI - A new paradigm for leaders in medicine. PMID- 14520783 TI - The evolution of palliative care. PMID- 14520784 TI - Dame Cicely Saunders. PMID- 14520785 TI - The gift of nothing. PMID- 14520786 TI - What I learned while dying. PMID- 14520787 TI - Worthy to serve the suffering revisited. PMID- 14520788 TI - Parallels and paradigms. PMID- 14520789 TI - Combat. PMID- 14520790 TI - Journey through illness. A perspective on the patient-physician relationship. PMID- 14520791 TI - A promise not kept. PMID- 14520792 TI - Billroth and Brahms. A musical friendship. PMID- 14520793 TI - [Acute dysphagia: often there is a readily treatable cause]. AB - Three patients, one woman aged 45 years and two men aged 40 and 62 years, presented with acute dysphagia due to oesophageal obstruction by a piece of food. In the woman symptoms of oesophageal perforation developed after the piece of food was removed by rigid endoscopy; she recovered after treatment with a stent, antibiotics and acid inhibitors. The younger man had a stricture of the oesophagus that was dilated. The older man had a Barrett's oesophagus and also oesophagcal adenocarcinoma; he was free of symptoms three years after resection of the oesophagus and the creation of a tubular stomach. In adults, 60% of acute oesophageal obstructions are caused by food impaction, which is associated with a high incidence of secondary pathologic findings in the oesophagus (75-97%). Evaluation of the oesophagus by flexible endoscopy contributes to an adequate diagnosis. Moreover, it can be used to treat the cause of the obstruction. If dilation therapy is started early after detecting a benign stricture in the oesophagus, it reduces the likelihood of recurrence. The detection of early-stage oesophageal malignancies may improve the prognosis of patients with this disorder. PMID- 14520794 TI - [Diseases and the diseased--scientific and alternative medicine]. AB - Exposing alternative medicine as quackery and unscientific is necessary, but one should be aware of the limitations of scientific medicine as far as the care of individual patients is concerned. Medicine is primarily about people, not about science. Current regular medicine uses a rational approach and carries out actions of which the effectiveness has been demonstrated in controlled trials. But some questions cannot be translated into a testable hypothesis, e.g., what is the best treatment for the individual incurable cancer patient sitting in my surgery right now? However, this question is a relevant one, especially for the patient involved. In daily clinical practice, there is no clear distinction between scientifically-based and not-scientifically-based acts. The task of medicine is to cure, but also to encourage and to console. When exposing alternative medicine, one should be able to offer the patient a different approach; otherwise the exposure is nothing more than a nihilistic exercise. Alternative medicine does not cure diseases, but it sometimes helps diseased people. PMID- 14520795 TI - [Hemopyrrollactamuria (HPU); from spots to pseudo-disease]. AB - In recent years, patients in the Netherlands confront their doctors with the diagnosis 'haemopyrollactamuria' (HPU), based on the presence of the haemopyrrollactam complex in their urine. The diagnosis is made by a commercial laboratory in the Netherlands (www.keac.nl). We have not been able to find peer reviewed scientific literature on this metabolic disease. The haemopyrrollactam complex represents the so-called mauve spot, which was the subject of much controversy in schizophrenia research in the previous century. Reviewing all of the available data, we feel that HPU should be classified as a pseudo-disease. PMID- 14520797 TI - [Enfuvirtide, the first representative of a new class of drugs for the treatment of HIV infection : HIV fusion inhibitors]. AB - In March 2003, enfuvirtide was approved in the USA and the European Union for the treatment of patients with HIV infection who have experienced failure or intolerable side effects of treatment with at least one representative of each antiretroviral drug class. Enfuvirtide has a new mode of action: it binds to the viral envelope glycoprotein 41 that is involved in the fusion of the virus to the membrane of the CD4 T cell. In two large phase III studies, 90 mg of enfuvirtide administered twice daily subcutaneously in addition to a background treatment of other antiretroviral drugs, had a significant favourable effect on both the plasma viral load (decrease) and the CD4 counts (increase) compared to the background treatment alone. Disadvantages of treatment with enfuvirtide are its subcutaneous administration (98% of the patients had local adverse reactions) and the high costs involved (1500 euro per patient per month). PMID- 14520796 TI - [Simplified diagnosis of deep venous thrombosis by applying clinical score and D dimer concentration]. AB - Serial compression ultrasonography of the leg is a safe but inefficient method of diagnosing deep venous thrombosis (DVT). The introduction of the clinical score and the D-dimer test heralds a change in the diagnostics of DVT. It is now safe to withhold anticoagulation from patients following one single normal ultrasonography in combination with a low clinical score or a normal D-dimer concentration. Recent studies show that DVT can safely be excluded on the basis of the combination of a normal D-dimer concentration and a low clinical score alone. PMID- 14520798 TI - [Diagnostic image (155). A man with facial emphysema. Subcutaneous facial emphysema due to dental cleaning]. AB - A 47-year-old man developed subcutaneous emphysema of the right side of his face during a dental cleaning procedure by the dentist. PMID- 14520799 TI - [Less use of alternative treatments by cancer patients in 2002 than in 1999]. AB - OBJECTIVE: To determine the use of alternative diets and other alternative treatments in 2002 compared to 1999. DESIGN: Descriptive, questionnaire. METHOD: During the period 13-26 May 2002 a survey was held among all patients visiting the outpatient clinic of the Netherlands Cancer Institute/Antoni van Leeuwenhoek hospital, Amsterdam, the Netherlands. Patients were asked about their current and past use of alternative therapies, their reasons for using these therapies, the way they were informed about these therapies and the expenses involved. The data were compared with the results of a similar study during the period 15-19 March 1999. RESULTS: Of the 729 patients who fulfilled the inclusion criteria, 66 (9%) declined to participate in the study. Of the remaining 663 patients (average age 58.5 years; 28% male), 131 (20%) used an alternative therapy. Of these, 43 patients (7%) used an alternative diet, mainly the Houtsmuller diet, and 88 patients (13%) used a mixture of alternative therapies such as homeopathy, vitamins and herbs. In 1999, 131 patients (30%) used an alternative form of treatment, 51 (13%) of whom used a diet. Of the 43 users of diets in 2002, 11 (26%) believed that the diet would slow down the disease process; in 1999 this was 53% (27/51). Of the 131 users of alternative therapies in 2002, 55% had been made aware of the possibilities of alternative treatments via family and friends. Internet and TV played a minor role as a source of information. 33 (79%) of the diet users informed their physician or nurse about the use. The diet users spent an average of 170 euro per month on their diets. CONCLUSION: Both the percentage of cancer patients who used an alternative diet and the percentage of diet users who believed that a diet could affect the course of the disease were reduced by half compared to three years earlier. PMID- 14520800 TI - [Results of in vitro fertilization in Dutch subfertile couples in Ghent, Belgium, 1993-2002: no patient group with decreased prognosis]. AB - OBJECTIVE: To describe the results of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) of Dutch patients who came to Ghent University Hospital (GHU), Belgium, for treatment, during the period 1993-2002. DESIGN: Retrospective data collection, description and analysis. METHOD: Dutch and Belgian patients were compared with respect to: age, indication for treatment, duration of infertility, stimulation protocol, number of ampoules of gonadotrophins, duration of stimulation, number of follicles, number of oocytes, number of fertilized oocytes, number of transferred embryos, quality of transferred embryos, number of frozen embryos, pregnancy, outcome of pregnancy, duration of pregnancy, sex and weight of the child or children. RESULTS: In the period 1993-2002 (10 years), 6099 treatment cycles were carried out in Belgian and 4801 in Dutch couples in the GHU. The duration of infertility was longer for the Dutch than for the Belgian couples and the Dutch women were on average three years older than the Belgian. The male infertility rate and proportion of ICSI cycles was higher for the Dutch than for the Belgian couples. In 17% of the cycles surgically retrieved sperm was used because of azoospermia. The numbers of follicles, oocytes retrieved, as well as the number of fertilized oocytes were all larger for the Belgian than for the Dutch patients. An equal number of embryos was transferred in both groups (2.24). The ongoing pregnancy rate per cycle was a little lower for the Dutch (23.2%) than for the Belgian couples (25.3%). The percentage multiple pregnancies was not different (27.8% for the Belgian and 25.1% for the Dutch patients). The Dutch singleton babies were born six days later than the Belgian, but the children differed in no other way. From 1996 to 2000 the ongoing pregnancy rate was 23.8% per first cycle for Dutch patients treated in the UHG, as compared to 19.4% in the Netherlands. CONCLUSION: Dutch patients treated in the GHU differed from Belgian patients in that they had a longer duration of infertility and a longer history of previously failed treatments. Although the pregnancy results were slightly lower for Dutch than for Belgian patients, this does not seem to be clinically very relevant, especially not in view of the overall Dutch national results. PMID- 14520801 TI - [Arterial hypoxemia due to packing of the nose]. AB - In a 75-year-old man who had experienced a serious nosebleed, as a consequence of collapsing, the nose was packed bilaterally. An evaluation of the dyspnoeic complaints revealed abnormal blood gas values which indicated hypoxemia. Ventilation perfusion scintigraphy and a lung function test did not reveal any abnormalities. Prior to additional investigations, the packing was removed and then the complaints disappeared and the blood gas values returned to normal. That nose packing can lead to arterial hypoxemia, with or without hypercapnia, has already been described. The underlying mechanism is probably determined multifactorially and is not completely clear. Control of the oxygen saturation seems to be indicated for patients with packing, and nasal obstruction should be considered in the differential diagnosis of hypoxemia. PMID- 14520802 TI - [Medical-scientific societies, quality control and disfunctional physicians]. AB - One doctor denied an existing fatal malignancy and diagnosed cancer in patients who did not have it. He and two other doctors gave the impression that they could cure patients by methods of which the efficacy has not been demonstrated. All three were members of a society for alternative medicine; two of them no longer are. The third, an internist, is still a member of his alternative scientific medical society, but of the Netherlands Association of Internal Medicine as well. A clinical chemist with a private laboratory that continually diagnoses nutritional deficiencies in a disputable way, and a neurologist who cooperates with a highly controversial American back physician were not expelled from their associations nor corrected by them. The Dutch Health Inspectorate is investigating dysfunctioning physicians and may well take action against them. The medical-scientific societies could well pay closer attention to their task of promoting the quality of the professional activities of their members. PMID- 14520803 TI - [The press as a vehicle for the introduction of a fashionable disease]. AB - Two journalists wrote a book on HPU (haemopyrrol-lactamuria), a hitherto unknown metabolic disease that was alleged to cause a whole range of complaints. The book is based on the ideas of a biochemist who is also the director of a commercial organisation that offers a diagnostic test as well as a therapy (nutrition supplement) for HPU. The release of the book was accompanied by a press release that provoked various publications in both biomedical journals and the lay press. In response to these reports, the director sometimes registered a complaint with the Dutch Council for Journalism, but on other occasions he did not. The content of this book is in no way based on scientific research. In retrospect, it is obvious that the authors, the biochemist and the publishing company have joint commercial ties. All professional standards of scientific research and journalistic decency are thus being violated. In this way, they are doing harm to the independence and integrity of researchers and journalists in the public image. All the activities of the press seem to have been an essential part of a deliberate marketing strategy for the introduction of a non-disease. PMID- 14520805 TI - Activist Douma, RN and Mayor, speaks out. Interview by Sandy Marron. PMID- 14520804 TI - [Biosensors for continuous glucose and lactate monitoring]. PMID- 14520806 TI - The value of clinical experience. PMID- 14520807 TI - Two examples: trust your clinical hunches. PMID- 14520808 TI - Computers in nursing. PMID- 14520809 TI - Female foeticide on the rise in India. PMID- 14520810 TI - Impact of self-instructional module for the nurses on nursing-management of the patients having chest tube drainage. PMID- 14520811 TI - Don't be fooled. PMID- 14520812 TI - World of difference. PMID- 14520813 TI - Out and about. PMID- 14520814 TI - Learn to read the signs. PMID- 14520815 TI - Coming soon ... CHAI. PMID- 14520816 TI - Fighting back. PMID- 14520817 TI - Take control. PMID- 14520818 TI - Do you really listen? PMID- 14520819 TI - Waiting on yellow. PMID- 14520820 TI - Family-witnessed cardiopulmonary resuscitation. AB - BACKGROUND: How nurses care for patients who have a cardiac arrest and for their family members who witness it has been identified as being paramount in determining relatives' acceptance of death and their ability to cope thereafter. In this article the author analyses the available literature, focuses on the advantages and disadvantages of families being present during cardiopulmonary resuscitation (CPR) in hospital and questions what effect this has on the grieving process (Kubler-Ross 1970). CONCLUSION: Nurses should take into account the views of relatives, although attempted resuscitation should never be compromised by family members being present. Evidence suggests that it can improve the grieving process for families and be helpful to patients who survive. PMID- 14520821 TI - Who is best qualified to teach bioscience to nurses? AB - Since the professions moved into higher education, diversity has developed in the amount, depth and method of bioscience teaching in nursing and midwifery courses. Bioscience encompasses biology, life science, anatomy and physiology. This diversity is a cause for concern at a time when nurses and midwives are taking on more of the traditional medical tasks such as prescribing and running clinics. Students need to acquire a sound grasp of anatomy and physiology and to achieve this a substantial amount of curriculum time needs to be devoted to bioscience. The main argument concerns not what should be taught but who should teach bioscience to students; whether this should be specialist lecturers from higher education science departments or nursing and midwifery teachers from health studies. This article makes the case for collaboration involving subject specialists and nursing/midwifery teachers and this is illustrated by examples of how such collaboration works in one higher education institution to produce a practical laboratory-based course. The conclusion is that time spent in life science laboratories should not be considered a waste of nursing/midwifery teaching time because the life science laboratory is a microcosm of clinical practice. This relevance can be emphasised through collaboration between nursing and bioscience lecturers. PMID- 14520822 TI - The role of nutrition in diabetes. AB - The author provides an overview and update on the current nutritional recommendations for diabetes and advice on methods of putting theory into practice. PMID- 14520823 TI - Bone up on osteoporosis. PMID- 14520824 TI - General prescribing principles. Supplementary prescribing: an overview. AB - The independent prescriber (doctor) is responsible for the diagnosis and setting the parameters of the clinical management plan. The supplementary prescriber decides on the product, dosage, frequency, and other variables within the confines of the management plan. The patient's progress must be regularly reviewed by the independent prescriber. The independent prescriber may at any time review the patient's treatment and/or resume full responsibility for the patient's care. The independent and supplementary prescriber must share a common patient record. There must be a good working relationship between the prescribers. Nurse prescribers need to be clear at all times which prescribing regime they are working under--district nurse/health visitor prescriber, extended formulary nurse prescriber or supplementary prescriber. PMID- 14520825 TI - Preparing for a disaster. Healthcare providers try to ready themselves for biological, chemical attacks without the benefit of government funding. AB - A new era of security alerts and biological terrorism has descended on the nation. Since Sept. 11, 2001, hospitals have been busy preparing for the scenarios of possible terrorist attacks. But providers say federal funding has fallen far short of what is needed to achieve their goals. At left, staff at Beth Israel Medical Center-Kings Highway Division in New York undergo emergency preparedness training. PMID- 14520826 TI - Legal hang-ups. JCAHO offers self-assessment options to avoid lawsuits. PMID- 14520827 TI - Losing it. HHS seeks to cut Tenet hospital from federal programs. PMID- 14520828 TI - 2-yard line. Bush may need to break logjam over Rx benefit. PMID- 14520829 TI - Rolling back. Calif. limits malpractice insurer's rate hike. PMID- 14520830 TI - Mixed reviews. Final EMTALA rules don't please everybody. PMID- 14520831 TI - End of an era. AHIMA to establish electronic record standards. PMID- 14520832 TI - On outcomes, the time has come. National database is needed to ensure the right care is given for each condition. PMID- 14520833 TI - Trial and error. Medical courts, arbitration systems are among the ideas gaining attention as answers to the malpractice liability crisis. PMID- 14520834 TI - Cost and complexity. Annual MGMA gathering to address issues weighing on medical practices. PMID- 14520835 TI - Medical advances. For many diabetics, development of oral insulin--whether via an inhaler or a capsule--could mean the end of daily needlesticks and the beginning of a simpler treatment regimen. PMID- 14520836 TI - [Classification of periodontal diseases]. AB - During the past centuries the basic understanding of periodontal disease and the concepts of etiopathogenesis have evolved and substantially changed. The changes in paradigm has always been followed by the conceptual changes in the classification of periodontal disease. The earliest entirely descriptive classifications were based on the clinical signs and symptoms of periodontal disease. These were followed by a classification system based on the classical pathology paradigm established by Orban and Gottlieb. This was replaced by the next classification reflecting the infection/host response paradigm during the late '60s started with the pioneering works by Loe and coworkers. It is because many data indicated that the host response (determined by genetics, immunology and behavioral factors) was at least as important in the clinical manifestation of periodontal destruction than the bacterial biofilm this classification had been also updated. The AAP published its latest official classification system in 1999. Nevertheless this new system is far from perfect and should evolve as soon as enough data are collected by periodontal research to answer many controversy still existing to day. PMID- 14520837 TI - [The beginnings of independent dental education at the Medical University of Budapest]. AB - Independent dental education in Hungary has been started in September 1952 at the University of Budapest. The previous system based on a diploma in general medicine + a postgraduate-specialist education has been stopped, although the claim on dentists has grown significantly in the country. The Dental Section of the Medical-Health Trade Union with the aim to solve this problem, accepted the establishment of an independent dental faculty within the Medical University of Budapest. The educational reform became realised in 1952 by an official regulation. The new class started to study at the Clinic of Stomatology, taught by the staff of the clinic. PMID- 14520838 TI - [Factors impeding obstructed teeth in their eruption. Case report]. AB - With preparation of six study cases the objective of the authors was to abrow colleagues' attention to the fact that teeth remained in impactio and retention might cause complaints not only due to normal anatomical reasons but some complications might occur resulting from anomaly of development. The authors consider the topic extremely important by reason of the case's varieties and the clinical observations made recently and frequently. An increased attention to this case is reasonable since eruption of teeth might happen in case of removal of obstructic fact in time, permanent row of teeth returns to normal, and adequate dental arch might develop. It is also obvious that medical treatment of impact abnormalities derived from anomaly has lack of exact therapy theories, only general protocol can be provided. PMID- 14520839 TI - [Time parameters of dental hypnosis]. AB - Dental treatments of high dental anxiety patients (n = 20) under hypnotic conditions were investigated. Time need of main hypnotic events like rapport; induction: deepening; calming: developing imagination; dehypnosis; discussion was measured. Total time was significantly higher (p < or = 0.05) than pure dental treatment time, but the time the patients percepted was significantly less then total (p < or = 0.05), even somewhat less then pure dental treatment time. Time distortion correlated significantly (negative Pearson's correlation; p < or = 0.05) with the time need of rapport (-0.50), calming (-0.51) and dehipnosis ( 0.46), and with the time need of developing delightful imagination (+0.57) (positive Pearson's correlation; p < or = 0.05). There was no significant correlation between time distortion and the time need of hypnotic induction, deepening, total time need of hypnosis, or pure dental treatment time. PMID- 14520840 TI - [Bone substitution and remodeling of the jaw bones. A review]. AB - Replacement of defects of the jawbones is one of the rapidly developing fields of maxillofacial surgery. The main criterion for the success of bone replacement is effective remodelling, i.e. the quantitative transformation of the inserted bone substitute material into bone of the same quality and the same structure as the original. The aim of this report is to survey the literature background of our knowledge relating to remodelling. An account is given of the mechanism of transformation of the bone-substitute material into bone, and of clinical, radiological and histologic methods with which the transformation can be appropriately followed. Remodelling is a protracted process, and conclusions relating to it can be drawn only from examinations performed at isolated points of time, which are often difficult to objectify. We may speak of complete remodelling if the inserted material can no longer be recognized, and only bone with a structure agreeing with that of its environment is to be seen in the augmented area. PMID- 14520841 TI - [Modification of the photo-acoustic stimulation in the psychotherapy of oral psychosomatic patients. Preliminary experiences]. AB - The possible use of a modified photo-acoustic stimulation technique was investigated in the explorative part of the psychotherapy of atypical oro-facial pain patients (n = 20; 13 females, 7 males; age between 39-59; psychiatric diagnostic categories in ICD-10: F41.2, F45.1) Since the stimulatory effects of this method cause difficulties to induce free imaginations, previously the main indication of this technique was the ego-strengthening of the patients, to help them to recognise and to verbalise emotions. In the modified technique, specific suggestions coming from detected changes of the patients' emotions, their symptoms, or their social activities are used. Our early results suggest that the modified technique advantageously speeded up the therapeutic process, but did not change the final therapeutic outcome. PMID- 14520842 TI - [How we treat patients with cardiovascular diseases in real clinical practice]. PMID- 14520844 TI - [Effect of a compound preparation pumpan on exercise tolerance and clinical course of ischemic heart disease]. AB - AIM: To study effects of the drug pumpan on exercise tolerance and myocardial ischemia induced by exercise in patients with ischemic heart disease of functional class II-III. MATERIAL AND METHODS: The study included 30 patients with various forms of ischemic heart disease (myocardial infarction, coronary artery bypass grafting) having stable effort angina of functional class II-III. RESULTS: The addition of a compound drug pumpan to a standard antianginal therapy reduced the number of anginal attacks and ST depression at bicycle exercise test. CONCLUSION: Pumpan is an adjuvant medication in conduction of antianginal therapy in ischemic heart disease patients with stable angina of effort of functional class II-III. PMID- 14520843 TI - [Efficacy and safety of the TAIS coronary stent implantation: nine month multicenter study]. AB - AIM: Assessment of clinical and angiographic results of a balloon-expandable steel matrix stent TAIS in patients with atherosclerosis of the coronary arteries with no history of previous interventions in the course of an open non-randomized multicenter trial. MATERIAL AND METHODS: The TAIS stent was implanted in 187 patients into 199 stenoses. 47% patients had clinical manifestations of unstable angina pectoris. 29% cases were complicated. The length of the stenoses reached 11.3 +/- 5.4 mm, 22% stenoses were prolonged. RESULTS: This coronary stenting was effective in 100% cases, neither acute nor subacute thromboses were seen. Myocardial infarction without occlusion of the stent developed in 3 patients. The incidence of cardial complications (death, angina, restenosis, repeated revascularization) in 6 month follow-up was 11.8%. An angiographic control in 6 months was made in 184 patients (194 stenoses). The vascular diameter loss index was 0.40 +/- 0.21, a restenosis level--10.7%. A logistic regression analysis has revealed significant correlations between the length of the stent and a target vascular diameter with subsequent restenosis. Repeated revascularization was performed in all the patients with stent restenosis. After control angiography cardiac complications developed in 13.9% patients. Nine month follow-up registered cardiac complications in 17.6% cases. CONCLUSION: The TAIS stent was found effective in prevention of thrombosis and restenosis in patients with a relatively high risk of intervention. PMID- 14520845 TI - [Co-renitek treatment of patients with moderate and severe forms of hypertensive disease]. AB - AIM: To evaluate efficacy and tolerance of a compound drug co-renitec combining an ACE inhibitor enalapril maleate and diuretic hydrochlorothiazide co-renitec taken for 16 weeks in essential hypertension (EH). MATERIAL AND METHODS: 28 patients with EH (16 males and 12 females aged 47-74 years) of mean duration 13.1 +/- 1.6 years. Blood pressure (BP) was monitored for 24 hours with the device SL 90207 (SpaceLabs Medical, USA). Microalbuminuria (MAU) was estimated with the use of immunoturbodimetric test. RESULTS: By 24-hour monitoring, co-renitec reduced day BP by 14.9/8.9 +/- 3/2 mm Hg, nocturnal BP lowered by 18.6/11.4 +/- 3/2 mmHg, pulse pressure also fell. Coefficient T/P was 53.5% for systolic BP (SBP) and 59.6% for diastolic BP (DBP). The target SBP was reached in 77% patients, target DBP--in 69%. Co-renitec significantly decreased MAU, albumines excretion normalized in 46% patients. CONCLUSION: Co-renitec lowers both day and nocturnal blood pressure, improves 24-h rhythm of BP, has a positive effect on the kidneys. This allows its recommendation as a first-line drug in patients with moderate and severe EH. PMID- 14520846 TI - [Effect of diroton on characteristics of 24-hours arterial pressure monitoring in hypertensive patients with polycythemia vera]. AB - AIM: To study the data of 24-h monitoring of blood pressure (MBP) and effects of an ACE inhibitor lisinopril (diroton) in hypertensive patients with polycythemia vera (PV). MATERIAL AND METHODS: 20 patients with arterial hypertension of degree II and III with PV aged 41 to 77 years. Mean duration of AH and PV was 11.8 +/- 2.2 and 2.0 +/- 0.2 years, respectively. Diroton was given as monotherapy in a single morning dose 10-40 mg for 4 weeks. 24-h MBP was made before the treatment and on the 4th week of the treatment. In addition to standard estimations, hour to-hour double product (DP) was estimated. RESULTS: After 4 weeks of diroton therapy there was a 12.2%, 9.5%, 25% and 15.4% fall in mean 24-h systolic blood pressure (SBP), diastolic blood pressure (DBP), pulse pressure (PP), DP, respectively. A positive effect was registered on variability and 24-h profile of BP. A response was achieved in 85% patients. The target level of mean 24-h blood pressure < 135/85 mm Hg was achieved in 65%, and 10% fall in SBD and/or DBP in 20% patients. ACE inhibitors' side effect--severe dry cough--was not encountered. CONCLUSION: PV aggravates arterial hypertension. Monotherapy with diroton effectively controls BP in hypertensive patients with PV in a 4-week course intake in a single morning dose and is well tolerated. PMID- 14520847 TI - [Clinical efficiency and tolerance of hydrogen sulfide balneotherapy in hypertensive patients living in arid zone]. AB - AIM: To examine clinical efficiency and safety of moderate hydrogen sulfide (HS) baths in the treatment of hypertensive patients living in arid zones. MATERIAL AND METHODS: Sixty patients with stable essential hypertension (EH) of stage I-II according to WHO classification living in arid climate took sparing HS baths. Efficiency of the baths was assessed by changes in blood pressure (BP), 24-h monitoring of BP. RESULTS: It is shown that systolic and diastolic BP after HS balneotherapy fell significantly both in daytime and at night. The 24-h profile of BP improved, heart rate decreased by 4.3%, physiological fluctuations of BP and BP variability were not damaged. CONCLUSION: Moderate HS baths are recommended for wider use in a hot climate of the arid zone as an effective and safe method of balneotherapy which can be adjuvant to basic medication of EH. PMID- 14520848 TI - [Use of clonazepam for treatment of patients with paroxysmal atrial fibrillation with regard to their psychoautonomic status]. AB - AIM: To study the efficacy of clonazepam in patients with paroxysmal atrial fibrillation (PAF) with reference to their psychovegetative status. MATERIAL AND METHODS: 50 patients with PAF of non-rheumatic origin entered the study group, 20 healthy examinees served control. All of them were examined using a clinicocardiological, cliniconeurological tests; their autonomic nervous system, emotional sphere, sleep disorders were studied before and after clonazepam treatment in a dose 0.5 mg 2 times a day for 30 days. RESULTS: A positive effect of clonazepam was noted in 20 (80%) patients with PAF, less frequent paroxysms occurred in 12 (48%) patients, in 5 patients they stopped, 12 (48%) patients tolerated the paraxysms better. The paraxysms became shorter in 10 (40%) patients, 10 (40%) patients felt better in the postparoxysmal period. A marked relief of psychovegetative syndrome, improvement of activity, mood and quality of sleep were noted in all the patients. CONCLUSION: Clonazepam is recommended for patients with PAF to improve antiarrhythmia therapy. PMID- 14520849 TI - [Effect of cordaron on systolic and diastolic functions of the left ventricle in patients with a mixed form of chronic low cardiac output and cardiac arrhythmias]. AB - AIM: To study effects of cordaron on central hemodynamics reflecting systolic and diastolic functions of the left ventricle (LV) in patients with a mixed form of chronic cardiac failure--a systolic dysfunction and LV diastolic dysfunction type I. MATERIAL AND METHODS: 14 patients with a mixed form of chronic heart failure (CHF) and cardiac arrhythmia were followed up for 6 months. All the patients received ACE inhibitors, diuretics, nitrates and, additionally, cordaron in a supporting dose 200-300 mg/day. Control of central hemodynamics was made with echocardiography before, 1, 3 and 6 months of therapy. RESULTS: For 6 months of therapy LV ejection fraction increased by 16%. LV diastolic function improved, primarily, due to a rise of the E max (by 52% for 6 months of therapy). This reflected early diastolic filling of the LV. Improvement of LV diastolic function was associated with heart rate decrease. CONCLUSION: Cordaron used in addition to ACE inhibitors, diuretics, nitrates improves both systolic and diastolic LV function in patients with a mixed form of CHF. PMID- 14520850 TI - [Efficacy and safety of different beta-blockers in patients with isolated systolic hypertension associated with diabetes mellitus and obstructive pulmonary diseases]. AB - AIM: To compare efficacy and safety of atenolol, methoprolol and bisoprolol, as the most usable beta-blockers, in patients with isolated systolic arterial hypertension (ISAH) and concomitant diabetes mellitus (DM) and/or chronic obstructive pulmonary disease (COPD). MATERIAL AND METHODS: Forty tow patients with ISAH and coronary heart disease, 30 patients with these diseases associated with DM and 32 patients with associated COPD were randomized into three groups. Group 1 received atenolol in a dose 25 mg twice a day, group 2--metoprolol tartrate in a dose 25-50 mg twice a day, group 3--bisoprolol in a single dose 5 10 mg/day. All the patients were examined before the treatment and in 8 weeks. Arterial pressure was assessed at 24-h monitoring (ABPM-04 unit, Mediteck, Hungary) and quality of life (QL) was estimated by DISS Disability Scale. In addition, blood glucose was measured in patients with concomitant DM, external respiration function (ERF) was studied before and after the treatment in patients with concomitant COPD. RESULTS: In all ISAH patients there was a significant fall of systolic arterial pressure and heart rate. 2 hours after intake of atenolol and methoprolol blood glucose lowered significantly in diabetics as well as peak volume expiration velocity in patients with COPD. In the bisoprolol group ERF and blood glucose in DM and COPD patients remained unchanged. Atenolol deteriorated QL by the "job" and "social life" scales, methoprolol--by the scale "job" while bisoprolol improved the above parameters. CONCLUSION: The results of bisoprolol treatment are better than those of methoprolol and atenolol treatment of hypertensive patients with concomitant DM and COPD. PMID- 14520851 TI - [Clinicomorphological assessment of budesonide efficiency in patients with bronchial asthma]. AB - AIM: to study clinicomorphological efficacy of inhalation glucocorticosteroid budesonide (benacort, Pulmomed, Russia) in bronchial asthma. MATERIAL AND METHODS: Twenty patients with bronchial asthma were treated with budesonide. RESULTS: A response to budesonide was manifest to the end of treatment week 1. Budesonide reduced frequency of acute asthma episodes and the need in inhalations of short-acting beta 2-agonists. The peak expiratory velocity (PEV) rose by 12% in three months, variability of PEV lowered by 13%, in 6 months by 21%, in 12 months by 31% compared to pretreatment values. In 12 months hypersecretion and thickness of basal membrane decreased. Three-month treatment also reduced eosinophil and lymphocyte epithelial count and cell density of stromal infiltrate in bronchial mucosa. In 12 months cell density of stromal infiltrate diminished. CONCLUSION: Bronchial asthma treatment with budesonide for 12 months reduces 24 hour rate of acute asthma episodes, the need in the disease exacerbations, improves functional indices of respiration but morphological composition of bronchial mucosa does not normalize completely this showing the necessity of longer budesonide administration. PMID- 14520852 TI - [Clinicomorphological changes of liver in atherogenic dyslipidemia and after treatment with statins]. AB - AIM: To evaluate clinicofunctional and morphological changes in the liver of patients with atherogenic dyslipidemia and in the course of treatment with statins. MATERIAL AND METHODS: A general clinical examination, biochemical hepatic tests, determination of blood lipid spectrum, markers of viral hepatites B, C, G, TT, ultrasound hepatic imaging of the liver of bile ducts, punch biopsy of the liver were made in 60 patients (38 females and 22 males) at the age of 39 to 70 years with atherogenic dyslipidemia (ADL). RESULTS: Biochemical tests showed high activity of transaminases (1.5-2 times higher than normal) in 16 patients, hyperbilirubinemia in 12 patients, dyslipidemia of type IIb-III in all the examinees. Ultrasound investigation of the liver has found fat dystrophy in 58, cholesterosis of the gallbladder in 32, cholelithiasis in 17 patients. Histological examination of the liver evidenced for marked fat dystrophy of hepatocytes, perihepatocellular and periportal fibrosis, moderate portal and periportal hepatitis. 22 of 54 patients treated with statins appeared to have morphological signs of drug damage to the liver. CONCLUSION: ADL patients' livers are characterized by signs of non-alcoholic steatohepatitis. In its presence stains treatment leads to development of drug-related hepatic disorder--statin hepatitis. Therefore, hypolipidemic drugs in non-alcoholic steatohepatitis must be used individually or alternative treatments should be tried. PMID- 14520853 TI - [Effect of combined psychocorrection on dynamics of pain syndrome and psychological status of patients with spinal osteochondrosis]. AB - AIM: To study effectiveness of a new psychocorrection method (combined use of neuroleptic trifluoperasine and psychorelaxation therapy using biological feedback) in rehabilitation of patients with long-term pain syndrome provoked by spinal osteochondrosis (SOC) to determine predictors of response to this method of psychocorrection. MATERIAL AND METHODS: 132 patients with SOC and associated pain syndrome were examined using pain assessing methods and psychological tests. RESULTS: Psychocorrection including neuroleptic significantly raises the efficacy of SOC treatment and relieves pain, hypochondric fixations and depressive disorders. The reduction of pain syndrome intensity in the course of psychocorrection correlates with attenuation of hypochondric disorders and inclination to fixation. As to predictors of the treatment effect, the strongest relief of pain in psychocorrection was achieved in patients with hypochondric and anxiodepressive disorders. CONCLUSION: Combined psychocorrection comprising minimal doses of neuroleptic trifluoperasine and psychorelaxation is effective in the treatment of SOC patients with long-term pain syndrome. PMID- 14520854 TI - [Use of venofer for iron deficiency correction in patients undergoing programmed hemodialysis]. AB - AIM: To study iron metabolism in patients on programmed hemodialysis (PH) in oral and intravenous administration of iron drugs; to compare clinical and financial results of using such drugs. MATERIAL AND METHODS: A two-stage trial studied iron metabolism in 158 PH patients on replacement therapy with erythropoetin. They received correction of iron deficiency with oral drugs (stage I) and venofer (stage II). RESULTS: The study of iron metabolism has found its deficiency in 2/3 patients receiving oral iron: absolute (48%) and relative (20%). Administration of venofer led to a 2-fold increase in the number of patients with normal iron metabolism. The target Hb and Ht were achieved in 2.5 times more patients than before venofer treatment. The dose of erythpoetin in such cases was reduced by 40%. Side effects were not observed. The week cost of venofer treatment per patient was lower by 22.5$ than the cost of treatment with oral iron drugs. CONCLUSION: Venofer correction of iron deficiency in PH patients is more effective both clinically and financially than use of oral iron preparations. PMID- 14520855 TI - [Therapeutic efficacy of imatinib mesylate (glivec) in chronic phase of myeloid leukemia]. AB - AIM: To evaluate efficacy and tolerance of glivek in chronic myeloid leukemia (CML) in patients who failed interferon-alpha (If-a) preparations. MATERIAL AND METHODS: 79 patients in a chronic phase of Ph + CML with hematological and cytogenetic resistance or intolerance of If-a. The response to glivec was assessed by achievement of a complete hematological remission and the cytogenetic effect (the degree of reduction of cell clone Ph+ in bone marrow). Tolerance and safety of the drug was studied by monthly standard clinicohematological tests. RESULTS: Not only a hematological remission (92.4%), but also partial (46.8%) or complete (27.8%) elimination of BCR-ABL +/- cells were achieved after 12 months of the treatment. Glivec was well tolerated. Hematological toxicity primarily as neutropenia and thrombocytopenia were observed in 54.4 and 42% patients, respectively. Neutropenia of the third degree which made impossible to continue the treatment was observed in 29.1% patients; throbocytopenia of the third degree was registered in 16.5% patients. Among most frequent non-hematological side effects there were moderate edema, nausea, leg muscle convulsions, weight gain, arthralgias, skin eruption. All the complications were transient, were managed in all cases with only a short-time discontinuation of glivec therapy. CONCLUSION: High activity of glivec at early stages of CML allows using this drug as a first line therapy in patients with CML. PMID- 14520856 TI - [Incidence and characteristics of complications in aplastic anemia patients treated with antilymphocytic globulin]. AB - AIM: To examine frequency and characteristic features of complications arising in administration of domestic antilymphocytic globulin (ALG) in patients with aplastic anemia (AA). MATERIAL AND METHODS: From 1980 to 2000 125 patients with AA were treated with ALG made in Russia. Two and three courses of ALG were performed in 25 and 8 patients, respectively. RESULTS: The treatment was complicated most often with allergic reactions (AR) observed in 42.4% patients, hemorrhagic syndrome (HS) and infectious complications (IC). Goat ALG was more allergic than rabbit ALG (46.2 vs 34.6%, respectively). If the second course was made with ALG of the same kind, AR occurred in 37.5% patients while ALG of the different kind produced complications only in 11.8% patients. HS was recorded in 37.3% patients, its frequency was unrelated to ALG kind and was not severe (71.2%). IC occurred in 16.5% patients, their occurrence was also unrelated to the ALG kind and were of the first degree in 63.6%. CONCLUSION: Domestic ALG often results in AR. To reduce frequency and severity of complications in ALG treatment, it is recommended to use long intravenous infusions (12-18 h) and change ALG kind in subsequent ALG courses. PMID- 14520857 TI - [Efficacy of thyroxine and potassium iodide treatment of benign nodular thyroid lesions]. AB - AIM: To study efficacy of thyroxine (TX) and potassium iodide (PI) in the treatment of benign nodular thyroid lesions (BNTL). MATERIAL AND METHODS: 118 patients with BNTL (colloid or colloid hypercellular as shown by thin needle aspiration biopsy, 'cold" or "warm" by scyntigraphy findings) were randomized into two groups: 59 patients were given thyroxin and the other 59 patients PI. The day dose of TX (75-150 mcg) was prescribed according to serum concentration of TTH trying to diminish it to 0.5 mIU/l and lower. PI dose was 200 mcg/day. Most of the patients were treated for 6 months. The response was evaluated with ultrasound investigation which measured thyroid volume, the size and number of the nodes in it before and in the end of therapy. The treatment was found effective if the dominant node decreased in size by 50% and more compared to pretreatment values. RESULTS: The size of the dominant node decreased by 50% and more in 14 of 59 (23.73%) patients on TX and in 20 of 59 (33.90%) patients on PI. Both TX and PI prevented growth of the dominant node size and number of the nodes in approximately 2/3 cases. TX was more effective in young patients (40.92 +/- 3.45 years) vs older ones (47.50 +/- 1.46 years, p = 0.047) and patients with colloid nodes. PI was more effective in patients with shorter node existence (3.93 +/- 1.21 and 8.59 +/- 1.74 months, p = 0.02). TX reduced thyroid volume from 20.42 +/- 1.69 to 15.18 +/- 1.30 ml (p = 0.001), PI--from 18.34 +/- 1.57 to 15.36 +/- 1.25 ml (p = 0.001). CONCLUSION: TX and PI can inhibit or prevent the growth of thyroid benign nodes in approximately 2/3 patients especially in young patients with colloid nodes (TX) and in short existence of the node (PI). PMID- 14520858 TI - [Treatment of patients with visceral candidiasis]. AB - AIM: To study effectiveness of antimycotic medication with flucostat in patients with visceral candidiasis. MATERIAL AND METHODS: Diagnosis of bronchopulmonary, gastrointestinal, urogenital candidiasis was verified microbiologically in 152 patients aged 20 to 78 years. Candidiasis was treated with flucostat: 50 mg and 150 mg capsules per os and solution (100-50 ml) intravenously by dropper. RESULTS: The response was achieved in 140 (92.1%) patients: cure--in 107 (70.4%), improvement--in 33 (21.7%), mycological sanation--in 125 (82.2%) patients. CONCLUSION: Cure of chronic diseases complicated by mycotic infection is not always achieved after mycotic sanation. Some symptoms of severe chronic pathology persist and require long-term symptomatic treatment. PMID- 14520859 TI - [Changes in adaptation-compensatory processes in the course of acute ischemic stroke (clinico-biochemical comparisons)]. AB - AIM: To study changes in some components of stress-realizing system (SRS) and stress-limiting system (SLS) characterizing compensatory mechanisms in acute disturbance of cerebral circulation (ADCC). MATERIAL AND METHODS: The activities of SRS and SLS were assessed in 94 patients by changes in the concentrations of glutamate, aspartate, acid phosphatase for SRS and gamma-aminobutyric acid (GABA) for SLS, respectively. RESULTS: After analysis of clinicobiochemical correlations the following clinical phases were singled out: subcompensation, moderate decompensation, severe decompensation and terminal. The phase conversion depended on quantitative changes of SRS and SLS activity. This enabled characterization of severity in certain time periods. Imbalance was revealed between SRS and SLS because of hyperactivation or hypoactivation of one of the above systems. CONCLUSION: Imbalance between SRS and SLS converts stress-syndrome from nonspecific link of adaptation into a nonspecific link of the disease pathogenesis. PMID- 14520860 TI - [Improved prognosis of generalized AL-amyloidosis in a patient with multiple myeloma treated according to the VAD protocol]. PMID- 14520861 TI - [Current approaches to the treatment of chronic obstructive pulmonary diseases]. PMID- 14520862 TI - [Diet of patients with chronic low cardiac output: solved and unsolved problems of nutritional support]. PMID- 14520863 TI - [Amiodarone-induced thyroid dysfunction (pathogenesis, diagnosis, treatment)]. PMID- 14520864 TI - [Meloxicam: intramuscular administration in rheumatology]. PMID- 14520865 TI - [Formation and diversity of parasitophorous vacuoles in parasitic protozoa. The Coccidia (Sporozoa, Apicomplexa)]. AB - Data on parasitophorous vacuole (PV) formation in host cells (HC) harbouring different intracellular protozoan parasites have been reviewed and critically analysed, with special reference to the main representatives of the Coccidia. The vacuole membrane (PVM) is the interface between host and parasite, playing a role in nutrient acquisition by the parasite from the HC. The PV phenomenon is regarded as a generalized HC response to the introduction of alien bodies (microorganisms), which eventually reflects the evolutionary established host parasite relationships at cellular, subcellular and molecular levels. Special attention has been paid to the existing morpho-functional diversity of the PVs within the same genera and species of parasites, and even at different stages of the parasite life cycle. The PVM is generally considered to derive from the HC plasmalemma, whose biochemical composition undergoes significant changes as the intravacuolar parasite grows. The original HC proteins are selectively excluded from the PVM, while those of the parasite are incorporated. As the result, the changed PVM becomes not fusigenic for HC lysosomes. For Toxoplasma gondii and other cyst-forming coccidia (Isospora, Sarcocystis), a definite correlation has been noticed between the extent of rhoptry and dense granule secrets released by a zoite during HC internalization, on the one hand, and the pattern of the PV that forms, on the other one. In T. gondii, tachyzoites, known to discharge abundant secrets, commonly force the development of PVs limited with a single unit membrane and equipped with a tubulovesicular network in the lumen. Unlike, bradyzoites known to be deficient in secretory materials trigger the formation of PVs with a three-membrane lining composed of the changed invaginated plasmalemma in addition to two membranes of endoplasmic reticulum. The two different types of PV harbour, respectively, exoenteric and enteric stages of T. gondii, the latter being confined to the cat intestine only. Unlike, all endogenous stages of the classic intestinal coccidia (Eimeria spp.) develop within PVs limited with a single membrane, with some invaginations extending into the PV lumen. Unusual PV patterns are characteristic of the extracytoplasmic eimerian coccidia (Cryptosporidium, Epieimeria) and adeleid haemogreagarines (Karyolysus). In cyst forming coccidia, the PVM is actively involved in tissue cyst wall formation, thus protecting the encysted parasites from recognition by the host immune system. All this strongly suggests that the PV is far from being an indifferent membraneous vesicle containing a parasite, but represents a metabolically active compartment in infected cells. Since all the coccidia are obligate intracellular parasites, the mode of their intimate interaction with the HC, largely accomplished via the PV and its membrane, is vital for their survival as biological species. PMID- 14520866 TI - [Electron microscope study of haemolymph cells of Tettigonia cantans (Orthoptera, Tettigonidae)]. AB - The haemolymph of Tettigonia cantans (Fuess) has been studied at the preimago and imago stages with both electron and light microscopic methods. PAS-negative granules were detected in histochemical reactions. On electronograms, one type of haemocytes was referred to as granulocytes recognized in haemolymph. In the cytoplasm of granulocytes, two types of granules were found: those of mitochondrial origin, and those being derivatives of the Golgi apparatus. Secret discharge is realized by the merocrine way. Four stages of granulocyte development are distinguished: the formation of granules and organelle development; the formation and accumulation of granules, active secretion, and cell destruction. PMID- 14520867 TI - [Morphological diversity of centromere regions in polytene chromosomes of blackflies (Diptera, Simulidae)]. AB - Karyotypes of more than 120 species of 33 genera of the Palearctic blackflies (Simuliidae) were studied on squashed acetoorcein stained preparations of salivary gland polytene chromosomes in larvae. In the course of evolution of the family, a significant complication was noticed in the morphology of centromere regions of polytene chromosomes. In plesiomorphic species, centromeres are not pronounced morphologically and the general picture does not differ from that of other bands and interbands of the polytene chromosome. In species with apomorphic characters, a distinct precentromeric heterochromatin appears, whose manifestation is responsible for morphological diversity of centromere zones in polytene chromosomes. They are represented either by conspicuous slightly thickened heterochromatic bands or by large amplified blocks of heterochromatin or puff-like structure, being considerably extended as a result of despiralization of precentromeric heterochromatin. There are species, which more commonly lack chromocentre and their chromosomes are separated. Some other species have ectopic contacts between pricentromeric heterochromatin. In some species, this heterochromatin is organized as a compact chromocentre. This has been found only in representatives of southern latitudes, most frequently in evolutionarily young species with narrow specialization. PMID- 14520868 TI - [The effect of ouabain on sensory neuron neurite growth in organotypic culture]. AB - Ouabain, a known inhibitor of Na+, K(+)-ATP, taken in a wide range of concentrations, was investigated in organotypic tissue culture of dorsal root ganglia cells of 10-11 day old chick embryos. Ouabain inhibited neurite growth in a dose-dependent manner. The Hill coefficient was defined as 1, and the Kp value was estimated as 1 x 10(-10) M. At inhibitor concentrations exceeding 1 x 10(-9) M, the growth of neurites was totally inhibited. It is assumed that Na+, K(+) ATPase may play an important role in regulation of the process of neurite growth in sensory neurones. PMID- 14520869 TI - [The influence of ammonium and nitrate of nutrient medium on ultrastructural organization and photosynthesis of callus cells in Glycine max L]. AB - We studied the influence of exogenic ammonium on the functional activity and ultrastructural organization of cells of the mixotrophic soybean callus (Glycine max L.). Ammonium available in the nutrient medium increased the chlorophyll content, accelerating the rate of photosynthetic O2 evolution per unit of biomass. The presence of ammonium in the medium promoted formation of the protein synthesizing system, which manifested itself as increased numbers of ribosomes, and thylakoids of chloroplasts, and higher electron density of the stroma in mitochondria and cytoplasm of mixotrophic cells. It has been concluded that the use of ammonium may lead to activation of protein synthesis, thus rising photosynthetic activity and favouring formation and development of membrane structures in chloroplasts. PMID- 14520870 TI - Co-influence of transgene expression in mammalian cells. Mutual influence of transgenes on their expression in mammalian cells. AB - It becomes increasingly clear that therapeutic gene delivery should provide not only for the sustained high level of gene expression but also, in most cases, for the regulated expression of transgenes as much as it occurs under natural conditions. Over the past few years a variety of different systems have been developed in order to regulate the amounts of transcribed RNA upon administration of exogenous agents, or in autoregulated manner. While efforts were focused on optimizing gene expression at the transcriptional level, other levels are still overlooked. In the meantime, regulation of gene expression is not restricted to transcription, but is also executed at the post-transcriptional level, i.e. mRNA stability, processing, transport, translation, protein stability, and modification. Codon usage is considered to be one of the critical factors that limit the expression rate of heterologous genes in different organisms at the posttranscriptional level. HIV-1 structural genes gag, pol, and env represent one of the most extensively utilized models for studying codon usage-mediated effects on transgene expression. In the current work we demonstrate that the codon content affects not only CMV-driven HIV-1 gag expression but also the expression of luciferase reporter gene transcribed independently from the SV40 promoter. The expression levels of both transgenes co-transfected into the human H1299 were inversely co-dependent. The observed phenomenon may be described as sequence independent post-transcriptional gene silencing, which reflects the existing limitation of transgene expression in mammalian cells at the post-transcriptional level. Optimization of the codon usage may provide for the additional level of regulation of transgene expression in gene transfer experiments in order to maintain the concentration of the protein at the therapeutic levels. PMID- 14520871 TI - [Use of the real-time RT-PCR method for investigation of small stable RNA expression level in human epidermoid carcinoma cells A431]. AB - Real-time RT-PCR using fluorescence dyes (e.g. SYBR Green I) is currently the most sensitive and precise method for investigation of RNA level and has long been widely used for absolute and relative quantification of mRNA in the cell. This highly sensitive method allows measurement of different type RNA level in the cell based on the kinetics of the corresponding double-stranded cDNA amplification. Upon its binding to the minor groove of double-stranded DNA, SYBR Green I dye increases its fluorescence about 100-fold, and this increase can be recorded even at early cycles of amplification. During the real-time RT-PCR procedure the level of amplified DNA is measured after every cycle of amplification, which permits to perform quantification at the cycles when amplification curve has not yet reached the "plateau" range and corresponds to the range of exponential increase in DNA amount. This approach makes it possible to avoid misinterpretation of data typical of conventional PCR methods "in the end point" and caused by a deficiency of one or more reaction components at the late PCR cycles. We applied for the first time real-time RT-PCR using SYBR Green I for the measurement of the class III genes RNA-product level, that is, small stable non-translated RNAs--ribosomal 5S rRNA, initiator transfer RNAiMet1, and Alu-RNA, synthesized by DNA-dependent RNA polymerase III. We investigated the level of 5S rRNA-, tRNA- and Alu-gene expression in the cell being in different states: with prolonged generation period, activated to proliferation, and apoptotic. The expression level was judged from the content of corresponding RNA products in the total cellular RNA. The used approach enabled us to find out the specific RNA share in the total cell RNA. Human epidermoid carcinoma cells A431 were used as a model for investigating class III gene expression level in vivo. These cells expose on their surface an abnormally large amount of receptors to epidermoid growth factor (EGF), and the result of EGF action on A431 cells depends on the growth factor concentration. Low concentrations of EGF (0.1 ng/ml) cause active proliferation of A431 cells, but its high concentrations (10-100 ng/ml) cause apoptosis in these cells. Besides, upon growing in serum-free media, A431 cells continue to proliferate, but by this extending the generation period to 48 h, against 30 h on growing in serum-containing media. Hence, A431 cells can serve as a useful model for investigation of specific gene expression level in cells being in different physiological states, in both slowly and actively proliferating cells, and in apoptotic cells. For successful use of real-time RT PCR in 5S rRNA, tRNAi(Met)1 and Alu-RNA level quantification, we optimized the amplification reaction conditions. We took into account that the share of each particular RNA in the cell may vary--the share of ribosomal RNA is high, tRNAi(Met)1--low, and Alu-RNA--very low. Moreover, the level of some small RNAs (e.g. Alu-RNA) can vary significantly in cells of different lines. This explains why the amount of cDNA, gained by reverse transcription of total cellular RNA, and the concentration of specific primers used for PCR were different in each case. We showed that the expression of different class III genes--5S rRNA-, tRNA- and Alu-genes, was not similarly regulated in response to external stimuli, causing prolongation of generation period, activation of proliferation and apoptosis. 5S rRNA level was practically the same in A431 cells both having prolonged generation period and being activated by EGF in low concentration, but in apoptotic cells this level dramatically fell about 8-fold. Alu-RNA level was equal in cells with prolonged generation period and in apoptotic cells, and increased about 2-fold in cells activated by EGF in low concentration. The initiator tRNAi(Met)1 level in cells activated by EGF in low concentration and in apoptotic cells was by almost two times higher than in cells with prolonged generation period. The data obtained testify that the real-time RT-PCR method using SYBR Green I yields highly reliable and reproducible quantification for the level of class III gene RNA-products--small stable RNAs (5S rRNA, tRNA and Alu RNA). Examination of each specific RNA level requires individual selection for the amplification reaction conditions: the amount of cDNA and primer concentration in the sample. This is primarily caused by different expression levels in some particular class III genes within the frames of the cells, and by different levels of some small stable RNAs (e. g. Alu-RNA) in different cell lines. Special attention must be paid to the internal control for discriminating between specific RNA levels in proliferating and apoptotic cells, as in the late apoptosis RNAs of most types are degraded (for example, mRNA of "house-keeping" gene for RPLP0 protein, used as a possible internal control in our experiments). As far as the applied approach allows estimation of a specific RNA share in the total cellular RNA, we propose to chose as internal control mRNA, whose share doesn't change during the total RNA degradation in apoptosis and thus, mRNA degradation is not selective (in relation to other type RNAs). In that way, the real-time RT-PCR method, which is currently the most sensitive and precise method for quantification of RNA in the cell, holds much promise for the investigation of not only different mRNAs, but also small stable RNAs, synthesized by RNA polymerase III. PMID- 14520872 TI - [Low efficiency of DNA repair system in mitochondria]. AB - Under the action of endogenous reactive oxygen species and exogenous chemical and physical agents, significantly more lesions occur in mitochondrial DNA (mtDNA) than in nuclear DNA (nDNA). However, the mechanisms of DNA repair in mitochondria are less efficient that in the nuclei. The mechanisms of nucleotide excision repair capable of removing UV-induced lesions or other complex adducts induced by chemical compounds are not operative in mitochondria at all. At the same time, mitochondria of some kinds contain a photoreactivation enzyme providing monomerization of cyclobutane pyrimidine dimers. Also, the enzyme system for DNA base excision repair (BER) and O6-alkylguanine-DNA alkyl transferase are functional in mitochondria. However, the rate of BER-controlled repair of lesions in mtDNA is lower than that in nDNA. The literature data suggest that the controlling system for the delay of DNA replication till the repair complexion (cell cycle checkpoint) cannot be provided in mitochontria. Besides, it remains unclear whether the mismatch repair mechanisms are operable in mammalian mitochondria. On the other hand, double-strand breaks in mammalian mtDNA are possibly repaired by involving the DNA-dependent protein kinase complex, and the process of BER is affected by poly(ADP-ribosyl)ation of proteins. Possible consequences of induction of the increased level of damage in mtDNA and the low efficiency of repair systems in mitochondria are discussed in this review. PMID- 14520873 TI - [Myocardium of mdx mice contains factor(s) that damage DNA structure and retard DNA reparation after gamma-irradiation (experiences in modeling system)]. AB - We studied the action of saline extracts of ventricle myocard (EM) of C57BL and mdx mice on DNA structure and repair of one-strand breaks of DNA in a modelling system. The system involves DNA repair in E. coli WP2 cells after gamma irradiation. Using standard technique, DNA reparation was estimated on measuring the speed of E. coli DNA sedimentation in alkaline sucrose gradients. It was shown, that EM of C57BL or mdx mice exerted no influence on DNA repair, which was completely declined within 60 min with EM present in the growth medium of permeabilized E. coli. Addition of C57BL mice EM into lytic solution does not accelerate DNA sedimentation of nonirradiated E. coli. At the same time, EM of mdx mice sharply accelerates DNA sedimentation of nonirradiated E. coli reducing DNA molecular weight from 200 x 10(6) to 135 x 10(6) Da. At entering in the lytic solution the EM of mdx mice also slows down E. coli DNA repair after gamma irradiation. It is supposed, that EM of mdx mice may contain a factor(s) damaging DNA in the E. coli lysate and presumably slowing down DNA reparation after gamma irradiation. Russian Foundation of Basic Research Grants 99-04-49390, 02-04-49870 and 00-04-49390. PMID- 14520875 TI - [Karyotype and morphology of Chironomus anthracinus Zett. (Diptera, Chironomidae) from eastern Kazakhstan]. AB - Larval morphology and polytene chromosomes of Chironomus anthracinus from eastern Kazakhstan have been described. Larvae have no lateral tubules on VII abdominal segment, but have paired ventral tubules on VIII abdominal segment. 2 n = 8 (A1.1 B1.1 C2.2 D1.1 E1.1 F1.1 G1.1). Two nucleoli are present in arms IVG and IIIF. Mapping of chromosomes, apart from antC2 arm, was performed after Shobanov (1996). B-chromosomes have been discovered in this species for the first time (in 19.4% individuals). Heterozygous inversions occurred in arms LA and IIIE in 46% of individuals. Compared to C. anthracinus from Yaroslavl Region, differences in morphology and karyology observed in this species from eastern Kazakhstan are related to habitation of the population examined in the deep-water Lake Markakol on the boundary of the area range. PMID- 14520874 TI - [Delayed biological consequences induced by gamma-radiation in human tumorigenic diploid line SK-UT-1B]. AB - The progeny of SK-UT-1B cells that survived gamma-irradiation with 4 Gy up to the 80th passage was examined. Descendants of irradiated cells lost p53 transactivation properties. Simultaneously, in the presence of nocodazole coordination between M and S phases was disrupted. Meanwhile, descendants of irradiated cells maintained the accurate spindle assembly checkpoint. These data suggest that p53 transactivation function may be required for coordination of M and S phases, rather than for spindle assembly checkpoint. Since it is known that p53 regulates both these processes on the basis of data obtained, we suggest that functions of p53 required for coordination of M and S-phases and for spindle assembly checkpoint are separated. Besides, the data obtained indicate that radiation-induced chromosomal rearrangements are associated with activation of DNA recombination process. PMID- 14520876 TI - [Chromosome 21 mosaicism. A review]. AB - Despite the great efforts contributed to studies on the trisomy 21 etiology, basic mechanisms of nondisjunction are still poorly understood. Even less is known about mosaic variant of trisonomy 21. In this paper, some problems of masaicism are considered: (1) estimation of the prevalence of mosaicism in patients affected with Down syndrome, (2) determination of population rate of parental mosaicism, (3) evaluation of the role of ovarian mosaicism in etiology of Down syndrome, (4) origin of the extra chromosome, (5) mechanisms of production of mosaic lines, (6) phenomenon of skewed sex ratio (female predominance, in fetuses particularly), (7) effect of maternal age. PMID- 14520877 TI - [Semi-centennial history of the nuclear matrix at the turn of the 21st century]. AB - In 1974, Berezney and Coffey described what they called the nuclear matrix (NM), thus ignoring our priority, since we had isolated and characterized virtually the same skeletal structure 25 years before this discovery. The presence of NM in the live cell was doubted, because of unsuccessful attempts to recognize it in vivo. NM comprises the lamina, extracted nucleoli and an intranuclear fibrogranular network. The internal matrix is very labile, its presence and abundance depending on methods of isolation, whereas the isolated NM can be revealed as granules 25 30 nm in diameter. As the state of the interchromatin space changes with varying in vivo conditions, temperature and methods of isolation, doubts cast upon the very existence of NM are to be regarded as hardly valid, and new progress in its study may be expected in the XXI century. PMID- 14520878 TI - [Hormonal signal system of the lower eukaryotes]. AB - The literary and the authors' own data on the structural and functional organization of hormonal signaling systems in the lower eukaryotes (yeasts, trypanosomes, ciliates, slide mold Dictyostelium discoideum) have been summarized and analysed. On the basis of a comparative analysis of the primary structures of signal proteins in the lower and higher eukaryotes (G-protein alpha-subunits, enzymes-cyclases-adenylyl and guanylyl cyclases) some possible pathways of the evolution of proteins are suggested. At the level of unicellular organisms, the main blocks of hormone-sensitive signaling systems of the higher eukaryotes were created. Moreover, signaling systems of the lower eukaryotes ar more invariant than these of the higher eukaryotes. It may be associated with the fact that of functional blocks, typical for signaling systems of multicellular animals, fungi and plants, were selected from the numerous variants of signaling system blocks of unicellular organisms. PMID- 14520879 TI - [Use of immunogold labelling technique for immunoelectron microscope localization of proteins in Drosophila polytene chromosomes]. AB - Using gold labeled antibodies, we developed and tested an immunoelectron microscope (IEM) method for detection of protein localization in Drosophila melanogaster polytene chromosomes. This method is based on procedures widely used for indirect immunofluorescent (IF) staining of salivary gland polytene chromosome squashes. The application of IEM was evaluated by using specific antibodies against proteins earlier localized in both decondensed (interbands and puffs) and compact (bands) regions of polytene chromosomes. In all the experiments, IEM and IF images for homologous chromosome regions were compared. When applied to regions of loose structures, IEM enabled us to localize, with high precision, signals in fine bands, interbands and puffs. There was a good correspondence between immunogold EM and IF data. However, there was no correspondence for dense bands: gold particles were distributed at their boundaries, while the entire bands showed bright fluorescence. This discrepancy probably resulted from a poor penetration of antibodies conjugated to gold particles in the tightly packaged structures. From the results obtained it may by concluded that the IEM method is advantageous for studying the fine protein topography of loose decompacted regions of polytene chromosomes. And this must be taken into consideration when protein localization in polytene chromosomes is performed. PMID- 14520880 TI - [Prometaphase aberrations leading to the nuclei restitution]. AB - The mechanism of restitution nuclei formation in male meoiosis of Triticum Agropyrum F1 hybrid and monosomic wheat line Milturum 553 has been studied. Visualization of division spindle with Navashin's routine fixationle distribution method accompanied with acetocarmine-coloured squashed preparations revealed that a block in the kinetochore spindle thread formation, i.e. the absence of chromosome association with the division spindle, is the basic reason of restitution in these genotypes. On this background, the prometaphase spindle in abnormal meiosis of these lines demonstrates a number of vivid and inordinate aberrations revealing some regularities of spindle structure in the normal plant cell. PMID- 14520881 TI - [Phenotype characteristic of the ionomycin-resistant CD4+ subset of the peripheral blood T-lymphocytes]. AB - The differential sensitivity of peripheral blood (PB) CD4+ T lymphocytes to the calcium ionophore ionomycin was investigated. Effect of ionomycin exerted on T cells was time- and dose-dependent. We have shown that resistant cells belonged to some distinct T cell subsets. The resting naive CD4+CD45RA+ T cells showed a little, if any, resistance to ionomycin treatment. The primed CD4+CD45R0+ memory T cells behaved similarly as did ionomycin-resistant (IR) cells. Although IR CD4+ T cells had a typical "memory" phenotype, some quantitative differences were found in expression of CD11a, CD28, CD29, CD62L and CD243 markers between PB CD4+CD45R0+ T cells and corresponding IR cells. PMID- 14520882 TI - [The mammalian centromere organization]. AB - Routine and recently obtained data on the pattern and functions of the mammalian centromeres and kinetochores have been reviewed. Several problems of kinetochore formation (centromere recognition, anaphase checkpoint) are specially discussed, in addition to the role played by centromere DNA in the interphase nucleus consideration. PMID- 14520883 TI - [Beta-1 and beta-4 integrins, and laminin 67 kDa receptors in interaction between A431 cells and laminin isoforms ]. AB - Laminins, as basal membrane glycoproteins, are able to stimulate cell adhesion and migration, and to influence gene expression. The laminin molecule has a set of bioactive sites that interact with different integrin and nonintegrin receptors, and, as a result, the reaction of the same cell type to different laminin isoforms may be different. The aim of this study was to determine the contributions of both integrins with beta 1 and beta 4 chains and 67 kDa laminin receptor in the interaction of A431 cells with two laminin isoforms: laminin-1 and laminin-2/4. The obtained data show that integrin alpha 6 beta 4 is more specific for interaction with laminin-2/4 than with laminin-1 and takes part in the stage of attachment of A431 cells to laminin. 67 kDa receptor promotes cell spreading on laminin-2/4 and inhibits cell spreading on laminin-1. An assumption was made about the complex action of receptors for interaction of A431 cells with laminins ("integrin alpha 6 beta 4--67 kDa receptors" complex). PMID- 14520884 TI - [Effect of narrow fractions of chromatin non-histone proteins on the expression of membrane tumor-associated antigen MA-50 and phosphorylation of proteins of cultured rat hepatocytes]. AB - As earlier reported, the main component of narrow fractions of chromosomal non histone proteins (NHP) of kidney and of Zaidel hepatoma cells has its own protein kinase activity, and is identified as a heteroorgan NHP-antigen, which is intrinsic to the definite renal tissue and absent in the liver. Effects of narrow fractions of kidney and Zaidel hepatoma NHP on biosynthetic processes and sizes of hepatocytes were studied in vitro. It has been shown that as a result of a 5 h incubation of rat hepatocytes with a narrow fraction of renal NHP the proportion of small hepatocytes increases approximately by 12% as compared with that of cells cultivated without NHP. Besides, binding of organ-specific anti-kidney immune serum with a small hepatocyte population rises by more than 20%, which results from the expression of tumor-associated heteroorgan kidney-specific antigen on the hepatocyte surface. According to immunoprecipitation and subsequent electrophoresis, the molecular mass of a membrane heteroorgan antigen on the surface of hepatocytes amounts approximately to 65 kDa, and an active phosphorylation of cellular proteins takes place. The same effect on hepatocytes is produced by a narrow NHP fraction of chromatin of Zaidel hepatoma cells, whereas no phosphorylation is observed in the presence of liver NHP as well as in the absence of NHP. It is suggested that the heteroorgan NHP-antigen induces biosynthetic processes including synthesis of membrane tumorassociated antigen on the surface of hepatocytes cultivated in vitro by activation of cellular protein phosphorylation, which can lead to changes in size of cultivated cells. PMID- 14520885 TI - [Reactive oxygen species generation by external surfaces of aquatic organisms]. AB - We have demonstrated that water organisms of various phylogenetic levels--lower fungi, sponges, colenterates, molluscs, and fishes--produce reactive oxygen species (ROS) into the aquatic environment without any additional stimuli. Our experimental data support the idea that ROS production by marine invertebrates is calcium- and proteinkinase C-dependent process, whose functions still remain unknown. PMID- 14520886 TI - [Intracellular localization of XCAP-E protein in XL2 (Xenopus laevis) cells under normal conditions and during inhibition of pRNA transcription and processing]. AB - The interaction of condensin subunit XCAP-E with various nucleolar subcompartments in XL2 cells was studied. In the interphase cells, XCAP-E was associated with a granular component of nucleoli (as shown by double staining with antibodies against B23) and with small nucleolus-like structures in the nucleoplasm. Inhibition of transcription by actinomycin D does not disrupt interaction of XCAP-E with the granular compartment of segregated nucleoli. Treatment with DRB 5,6-dichloro-1 beta-ribofuranozide-benzimidazole causes disintegration of nucleolar fibrillar complexes, but does not affect nucleolar localization of XCAP-E. The data suggest that nucleolar association of XCAP-E is independent on the functional state of the nucleolus, and imply a possible role of this protein in rRNA processing and pre-fibosome assembly. PMID- 14520887 TI - [Induction of nuclear envelope formation around individual chromosomes under impact of hypotonic shock]. AB - In the present work we have studied the distribution of some proteins participating in the nuclear envelope assembly (lamins A/C, B and LAP2 alpha) in mitotic cells and after hypotonic treatment with 15% Hank's solution. In untreated cells, these proteins are localized in the nuclei of interphase cells migrate to the cytoplasm during mitosis. Hypotonic treatment of interphase, prophase and telophase cells does not lead to considerable relocalization of lamins A/C and B. However, unlike normal mitosis, in prometaphase and metaphase cells their chromosomes acquire affinity to lamins and LAP2 alpha. Comparative analysis of lamins and LAP2 alpha distribution have revealed that chromosomes have special sites for binding with different proteins. PMID- 14520888 TI - [Genetic variation in two forms of the common spadefoot toad Pelobates fuscus (Pelobatidae, Anura, Amphibia) distinguished by genome size]. AB - Genetic differences (presumed 23 loci) between two cryptic forms of Pelobates fuscus, differing in genome size, were examined by means of polyacrylamide-gel electrophoresis. This method allowed to discriminate between these forms. Average genetic distance (DN) between both the forms was equal to 0.311, ranging from 0.055 to 0.563. As a rule, differences within these groups were smaller (0.021 0.388). The data show obvious genetic differentiation between these two cryptic forms of P. fuscus. Differences between these forms and P. syriacus were significantly higher (in average 0.943). Genetic distances in relation to speciation and species concepts are discussed. PMID- 14520889 TI - The spore wall and polar tube proteins of the microsporidian Nosema grylli: the major spore wall protein is released before spore extrusion. AB - Incubation of Nosema grylli spores in alkaline--saline solution (10 mM KOH, 170 mM KCl) leads to solubilization of the major spore wall protein of 40 kDa (p40). Both the compounds of this solution are crucial for p40 solubilization. After spore incubation in 170 mM KCl no proteins were released in the medium. In contrast, 10 mM KOH causes a release of many spore proteins but only a small amount of p40. A long storage of spores (over a year) in water or 0.02% sodium azide results in a sharp decrease of p40 content. Specific polyclonal antibodies were obtained by immunization of rabbits with isolated p40. The specificity of serum was confirmed by immunoblotting. IFA showed reliable reaction on the envelopes of sporonts and sporoblasts, whereas only part of spores reacted with antibodies. This distinction may be due to changing surface antigens during spore maturation. Solubilization of p40 under alkaline conditions could be associated with spore extrusion, since a subsequent transfer of spores to neutral solution leads to their discharge. Subsequent wash of discharged spores with 1-3% SDS, 9 M urea and treatment by 100% 2-ME result in solubilization of protein of 56 kDa (p56). The maximum concentration of 2-ME is important for isolation of pure p56. Evidence has been provided that p56 is a protein of N. grylli polar tubes. Treatment of discharged spores by 2-ME in the presence of SDS results in solubilization of four additional proteins with molecular weights about 46, 34, 21 and 15 kDa. PMID- 14520890 TI - Evolution in the treatment of atopic dermatitis: new approaches to managing a chronic skin disease. PMID- 14520891 TI - An overview of atopic dermatitis. AB - Atopic dermatitis (AD) is an inflammatory disease, characterized by marked pruritus, which typically assumes a chronically relapsing course. Although AD is primarily a childhood disease, 60% to 70% of adults with AD suffer from hand eczema. Caring for patients with AD is time consuming and difficult. Patients and parents need proper education and detailed instructions from nurses and physicians to avoid flares and complications. Recognizing signs of infection and beginning appropriate treatment for them are important. Instruction on corticosteroid therapy as well as use of topical immunomodulators is crucial to effective acute and long-term management of AD. PMID- 14520892 TI - The role of the dermatology nurse in atopic dermatitis management. AB - The dermatology nurse plays a unique role in managing atopic dermatitis (AD). Education and prevention are the primary focus of the dermatology nurse, and he or she can also play a vital role as a patient advocate. PMID- 14520893 TI - Treatment options and new therapeutic approaches in atopic dermatitis. AB - Many patients with atopic dermatitis feel that their treatment is ineffective and 25% may be noncompliant with treatments involving corticosteroids. Developing safe and effective pharmacologic alternatives to steroids has presented a challenge to physicians and drug researchers. A new class of nonsteroid immunomodulators, topical calcineurin inhibitors, overcome some of the difficulties and concerns associated with steroid use. The introduction of new drug therapies reinforces the importance of dermatology nursing in managing atopic dermatitis. PMID- 14520894 TI - [Guanidine derivatives as reversible inhibitors of cholinesterases of various origins: conformation and complex forming]. PMID- 14520895 TI - [Activity and thermostability of acid phosphatase in Amoeba proteus, cultured at different temperatures]. PMID- 14520896 TI - [Regulation of adenylate cyclase system of Tetrahymena pyriformis by hormone and non-hormone agents and its dependency on adenylate cyclase basal activity]. PMID- 14520897 TI - [Insulin-like peptides of cerebro-pleural ganglia of the mollusc Anodonta cygnea: isolation, purification and radioligand analysis]. PMID- 14520898 TI - [Phenoloxidase system expression using chito-oligosaccharides as a immunomodulators in insects]. PMID- 14520899 TI - [Activation of the Na/H exchange in erythrocytes of the frog Rana ridibunda]. PMID- 14520901 TI - [Effect of adrenaline and piracetam on feeding behavior in fishes]. PMID- 14520900 TI - [Inositol- and galactose binding lectins from chicken brain fractions enriched with glial cells]. PMID- 14520902 TI - [Lithium transport in kidney of the gull Larus argentatus: induction of the Li+ renal netto- secretion by NaCl loading ]. PMID- 14520903 TI - [Sex differences in the reactivity of the sympathoadrenal system during ontogenesis of rats]. PMID- 14520904 TI - [Age-related differences of anxiety level in male and female rats during elevated plus-maze test]. PMID- 14520906 TI - [NO-synthase localization in gonad of freshwater bivalve mollusk Nodularia vladivostokensis]. PMID- 14520905 TI - [Peripheral blood leucocyte picture of green frogs under anthropogenic load]. PMID- 14520907 TI - [Functional asymmetry of the brain and its evolution in Humankind history]. PMID- 14520908 TI - Constitutionality of malpractice non-economic damage cap. PMID- 14520909 TI - New report clarifies the extent of the Medicaid shortfall. PMID- 14520910 TI - Large radiology practice focuses on patients and policies. PMID- 14520911 TI - Is Armageddon a reality? The question gets answered at scientific meeting. PMID- 14520912 TI - Thinking about adding "MBA" to your credentials? Here are some options. PMID- 14520913 TI - Protecting our antibiotic lifeline, Part II. PMID- 14520914 TI - Influenza vaccine ... to whom and when? PMID- 14520915 TI - Nutritional supplementation: new findings that counter conventional wisdom. PMID- 14520916 TI - Just what does "managed care" manage? PMID- 14520917 TI - Getting back in the race. Retired healthcare executives are being lured back into the office as organizations look for experienced leaders to take the reins. AB - All the young whippersnappers better look out--the veterans are coming back to show how it's done. More and more as of late, executives who chucked the 80-hour workweek grind are coming out of retirement to take the helm at beleaguered healthcare systems. MedCath Corp. board Chairman Steve Puckett, at left, says experience counts for a lot when handing over such responsibility. PMID- 14520918 TI - Either way ... you pay. Employers push big copays for hospital care. PMID- 14520919 TI - Either way ... you pay. Medicare HMOs seek more out-of-pocket payments. PMID- 14520920 TI - United they stand. Hospitals work together to offer malpractice coverage. PMID- 14520921 TI - Back to basics. Minn. Blues to abandon hospital ownership. PMID- 14520922 TI - Incremental progress. Conference committee supports reimbursement hike for critical-access facilities. PMID- 14520923 TI - Period of reconciliation. House, Senate seek common ground on spending bills. PMID- 14520924 TI - Trying to keep their own. Mentoring program aims to improve nurse retention. PMID- 14520925 TI - Under investigation. Hospitals may be liable for records sales. PMID- 14520926 TI - 2003 up & comers. Modern Healthcare and Witt/Kieffer present this year's class of rising stars in healthcare management. PMID- 14520927 TI - By the numbers. Top 15 executive recruitment firms. PMID- 14520928 TI - Blast injury. AB - Individuals with a blast injury are the victims most likely to be seen by any healthcare professional. An awareness of the potential injuries is important. Individual care should follow standards established for all trauma patients. PMID- 14520929 TI - Profiles in leadership. Robert W. Block, MD. PMID- 14520930 TI - Appendectomy at the time of cesarean section. AB - The current world literature estimates the incidence of appendicitis in pregnancy at approximately 1 case per 1500 normal deliveries. Observations of one physician and review of records from Saint Anthony Hospital in Oklahoma City have suggested potential benefit to routine examination of the appendix at the time of cesarean section and removal if it appears pathologic. The medical literature was reviewed regarding appendicitis in pregnancy and incidental or indicated appendectomy at the time of cesarean section. Many authors have described difficulties with diagnosis, as well as potential complications of appendicitis in pregnancy. Studies have confirmed the safety of performing incidental appendectomy at the time of cesarean section. The authors, therefore, propose that clinicians visualize and palpate the appendix at all cesarean sections, and remove those with evidence of inflammation or disease. PMID- 14520931 TI - Tulsa health care challenge: a compilation of issues and possible solutions. AB - The delivery of adequate health care to "all of the people" has become an increasing challenge in the United States. The problem of timely access to affordable health care is even more serious in Oklahoma. The University of Oklahoma-Tulsa Health Science Center, the OU College of Medicine-Tulsa, and Community HealthNET, Inc. have taken leadership roles in convening two Oklahoma Health Care Challenge summit meetings this past year. Tulsa community leaders are now addressing the seriousness of the local problem and, more importantly, are coming up with ways to alleviate it. The focus of the summits has centered on providing indigent care, which will provide relief to the Tulsa hospitals and physicians from the growing burden of charity care costs. The most beneficial results of these meetings have been a clearer definition of the problem specific to Tulsa, the recognition that the solution will require coalitions of multidisciplinary groups, and the realization that cooperation, as well as collaboration, is the key to success. PMID- 14520932 TI - Billing 101: the questions most often asked. PMID- 14520933 TI - Alcohol-related problems among adolescents. Part 2. PMID- 14520934 TI - Real men get the blues. Depression is twice as common among women as men, but it may be the guys who suffer most. PMID- 14520935 TI - An uncommon death. What to know about the vascular condition that killed John Ritter. PMID- 14520936 TI - [Current views on acquired valvular heart disease and the results of treatment]. AB - The article deals with the problems of diagnosis of acquired valvular defects (AVD) and indication to surgical treatment of AVD in accordance with international recommendations and modern knowledge of this issue. The following AVD diagnosis methods are described: heart auscultation, echocardiography, tests with regulated load, angiographic and radionuclide methods. Among pathological conditions, more attention is placed on stenoses, aortic, mitral and tricuspid valve insufficiency, IHD. Of importance is interpretation of the data obtained at diagnostic tests in terms of indications to cardiosurgical interventions, optimization of the terms and outcomes of the surgery. Statistics are provided on natural survival of patients and efficacy of surgical treatment. PMID- 14520937 TI - [Acute respiratory distress syndrome]. AB - Current literature on acute respiratory distress-syndrome (ARDS) is reviewed with characteristics of the terms "acute pulmonary lesion" and "ARDS". Clinical and x ray classifications of ARDS are presented. Etiology and pathogenesis, epidemiology, clinical manifestations and principles of current treatment are discussed. Use of mechanical ventilation of the lungs in ARDS patients is physiologically validated. Much attention is payed to application of natural and artificial surfactants. PMID- 14520938 TI - [The problem of biliary sludge]. AB - The authors review current literature on etiology, pathogenesis and classification of biliary sludge (BS); they analyse factors promoting formation of BS and show that ultrasonic investigation is the key diagnostic method, detecting various forms of BS and differentiating BS with parietal new-growths of the gallbladder, evaluating function of the latter and controlling efficacy of conservative therapy. Follow-up results demonstrate that BS can disappear spontaneously, be persistent in a part of patients, form stones in 20% patients. Clinical picture in BS has no specific symptoms but in 33-75% cases it is associated with development of biliary pancreatitis as a results of secondary dysfunction of the Oddi's sphincter. The latest findings on efficacy of litholytic therapy in BS and data on possible use of nonsteroid anti-inflammatory drugs as inhibitors of mucin production to prevent recurrent cholelithiasis are presented. PMID- 14520939 TI - [Clinical features of arterial hypertension regarding gustatory sensitivity to common salt]. AB - To specify clinical features and a course of arterial hypertension (AH) in respect of the amount of common salt (DS) and the threshold of gustatory sensitivity to DS, 134 hypertensive patients were examined. The patients having a high DS sensitivity are shown to take more DS and are characterized by more frequent cardiovascular and neurotic complaints, as well as by prevalence of anxiety and tension in their psychological status. Such patients more frequently have hereditary load of diseases; AH in them develops 10 years earlier, hypertensive crises occur more often. Left ventricular hypertrophy develops earlier, such risk factors as smoking and hypercholesterolemia combines often. PMID- 14520940 TI - [Indices of the lipid peroxidation--antioxidant defense system as markers of chronic cardiac failure in cardiomyopathies]. AB - Content of circulating products of lipid peroxidation (LPO), alpha-tocopherol (AT) and ceruloplasmin (CP), was studied in patients with hypertrophic and dilated cardiomyopathies (HTCMP and DCMP). The data obtained correlates with severity of chronic cardiac failure (CCF) and exercise tolerance of the patients. It is shown that the rise in LPO products level and simultaneous lowering of AT concentration in HTCMP and DCMP depend on the functional class of chronic cardiac failure by NYHA (1964). Content of lipoperoxides, AT and CP in the serum reflect the HTCMP patients' tolerance to exercise. Laboratory estimation of the levels of LPO products, AT and CP in patients with cardiomyopathies can substantially objectivize and supplement both clinical and functional-diagnostic assessment of CCF severity. PMID- 14520941 TI - [Spiral computed tomography with functional tests in diagnosis of chronic obstructive diseases of the lungs]. AB - The potential of spiral computed tomography (SCT) and high resolution computed tomography (HRST) in diagnosis of chronic obstructive pulmonary diseases (COPD) is described. Semiotic sings of bronchial asthma (BA) and chronic obstructive bronchitis (COB) are specified. Informative value of SCT in differential diagnosis of BA and COB including early stage of the diseases is analysed. An updated technique of SCT and HRCT of the lungs in COPD patients is presented. PMID- 14520942 TI - [Effects of dicetel on gastrointestinal motility and vegetative dysfunction in patients with duodenal ulcer]. AB - The great majority of patients with duodenal ulcer (DU) given a single oral dose (50 mg) of dicetel (pinaverium bromide)--a selective blocker of calcium channels- showed optimization of vegetative reactivity and vegetative support of physical and mental activities. This contributes to relief of psychovegetative syndrome of the modulation of dysfunctions of proximal gastrointestinal tract (GIT). The addition of dicetel to combined treatment of PH shortened healing of DU, relieved pain, stopped gastric and intestinal dyspepsia, corrected motor-tonic defects of the proximal GIT caused by dysfunction of different links of the regulatory adaptive system. PMID- 14520943 TI - [Enzymatic activity and therapeutic action of pepsin K in chronic gastritis with secretory failure]. AB - Pepsin K (chicken pepsin) in the test for hemoglobin proteolysis is 1.7 times superior to abomin (porcine pepsin) in the medium pH 2 and 44 times more potent in the medium pH 3. Even in pH 4 chicken pepsin retains its enzymatic activity while porcine pepsin in pH 4 looses its activity. These biochemical findings show that pepsin K is more effective in the treatment of enzymatic insufficiency of the stomach. Intake of 1-3 pepsin K tablets after meal for 4 weeks by 82 patients having chronic gastritis with secretory insufficiency was effective in 75% of patients. PMID- 14520944 TI - [Efficacy of antihelicobacter therapy in NSAID-induced gastropathies]. AB - The aim was to evaluate antihelicobacter therapy in the treatment and prophylaxis of NSAID-induced gastropathy recurrences in patients with rheumatic diseases (RD). 66 patients with RD (58 females and 8 males, mean age 53.6 +/- 12.6 years) participated in the study. They met the following criteria: the presence of ulcers or multiple (more than 10) erosions of gastric or duodenal erosions, administration of nonsteroid anti-inflammatory drugs, Helicobacter pylori (HP) in gastric mucosa biopsies. The patients were randomized into two groups matched by sex, features of RD, antirheumatic therapy, history of ulcer, GIT disease. All the patients received omeprazol in a dose 40 mg/day. Patients of group 1 took also amoxicillin (1 g/day) and klarythromycin (0.5 g/day) for 10 days. Group 2 was control. EGDS was conducted 2, 4 weeks and 6 months after the treatment. HP eradication and dynamics of morphological changes were examined in biopsies of the antral gastric mucosa and mucosa of gastric body. The efficacy of 4-week treatment (ulcer scarring and epithelization of erosions), rate of ulcer recurrences and erosions 6 months after treatment were compared. The efficacy of the treatment was 88 and 95% in group 1 and 2, respectively. HP eradication was achieved in 88% of patients of group 1. In 6 months, recurrences arose in 50 and 48.3% of patients of groups 1 and 2, HP was detected in 22% of patients of group 1. Morphological changes in group 1 patients was characterized by alleviation of chronic active gastritis. HP eradication does not raise the efficacy of the treatment and does not lower the risk of recurrences in patients with NSAID induced gastropathies. Active chronic gastritis is not the background for recurrences of NSAID-induced gastropathy. PMID- 14520945 TI - [The phenotypical symptom complex of connective tissue dysplasia in females]. PMID- 14520946 TI - [Psychovegetative features in patients with biliary dyskinesia]. AB - The aim was to study psychovegetative disorders in hypomotor and hypermotor forms of biliary diskinesia (BDK) in young patients. Psychological status and vegetative nervous function were studied in 62 patients with hypoBDK, 58 patients with hyperBDK and 33 control patients. According to AMPI, Spilberger's, Aizek's, Heck-Hess' tests, the psychological status of the patients from both groups was similar, while vegetative status in the hypoBDK group at rest and active orthostasis had a sympathetic trend. In hyperBDK group at rest sympathetic impacts enhanced, in stress these impacts intensified reaching higher values than in the control group. For the treatment of various forms of BDK, it is recommended to correct psychovegetative deviations with consideration of diskinesia forms. PMID- 14520947 TI - [Dynamics of a 24-h profile of arterial pressure and function of the vascular endothelium in nebivolol treatment of hypertensive patients]. AB - To assess a hypotensive activity of nebivolol (nebilet) including its action on the 24-h profile and baroreceptor control of arterial pressure (AP), vegetative regulation of the heart and vasculomotor endothelial function in AP patients, 30 AH patients were given nebivolol in a dose 5 mg/day for 8 weeks. The results of the treatment show that nebivolol effectively maintains a safe 24-h control of AP, acts positively on chronostructure of AP 24-h profile, produced no clinically significant side effects, corrects vegetative heart regulation and improves baroreceptor AP control. Moreover, nebilet modulates a stable vasodilating effect which improves function of vascular endothelium, initiates regression of left ventricular myocardium hypertrophy by raising its elasticity without changing much its contractile function. Thus, nebivolol is a highly effective drug for treatment of arterial hypertension which corrects a wide spectrum of unfavourable shifts in cardiovascular system function. PMID- 14520948 TI - [Psychological and psychosomatic problems of oncology]. AB - The paper concerns the issues of medical ethics and deontology in communication of a physician with cancer patients and their relatives. Ethic and deontological norms dictate the necessity to approach each patient individually, especially in terms of giving information about the disease. While informing the patient truly about his/her disease, it is important to keep the patient hoping and focus his/her efforts on radical treatment if the disease is not too advanced. If it is so, the patients need individual and group psychotherapy which is able to improve quality of life and to prolong the survival in cancer patients. PMID- 14520949 TI - [Implications of carbohydrate digestion time in diet therapy]. AB - The review is devoted to the problem of using information about dietary carbohydrate digestion in the practice of diet therapy of different diseases. Excessive consumption of carbohydrates, especially easily assimilable, entails disturbances not only in carbohydrate but also in fat metabolism. This increases the risk of development of many diseases including obesity, atherosclerosis and relevant sequelae, diabetes mellitus, etc. These diseases can be successfully treated by diet which changes structure and amount of diet carbohydrates. However, carbohydrate adsorption rate determined by glycemic index (GI) is underestimated. Now recommendations for diet therapy with application of GI are available only for patients with insulin-dependent diabetes mellitus though this index could be valuable for other diseases. A differential approach to the choice of carbohydrates-containing food with high, moderate or low GI would reduce the load on the insular apparatus and thus improve a clinical course of the disease. GI consideration in diet guides is necessary in practical activity of medical professionals. PMID- 14520950 TI - [Difficulties in diagnosis of glutenic enteropathy in the elderly]. PMID- 14520951 TI - [Primary gastric tuberculosis in a 81-year-old man]. PMID- 14520952 TI - [Problems of cardiac failure at the XXIV European Cardiology Congress (Berlin, August--September 2002)]. PMID- 14520953 TI - When hearts break. PMID- 14520954 TI - Love.com. PMID- 14520955 TI - CEO sufferings trickle down. PMID- 14520956 TI - Divining nature's plan. PMID- 14520957 TI - [Lytic enzymes of Trichoderma and their role in protecting plants from fungal diseases]. AB - Lytic enzymes of mycoparasitic fungi of the genus Trichoderma, capable of suppressing several fungal phytopathogens that originate in air or soil, are reviewed. The topics analyzed include (1) regulation of production of chitinases, beta-1,3-glucanases, and proteases; (2) molecular and catalytic properties of purified enzymes; and (3) their in vitro ability to degrade cell walls and inhibit sporulation or germ-tube elongation in various phytopathogenic fungi. Among the results summarized are reports of cloning the expression of genes coding for certain lytic enzymes of Trichoderma spp. These genes are used for obtaining plant transgenes with increased resistance to fungal diseases and Trichoderma transformants that produce higher levels of one lytic enzyme (a chitinase or protease) and thereby exhibit a more pronounced ability to suppress phytopathogenic fungi. PMID- 14520958 TI - [Inhibition of peroxidase oxidation of aromatic amines by substituted phenols]. AB - Peroxidase-catalyzed oxidation of o-phenylene diamine (OPD) was competitively inhibited by trimethylhydroquinone (TMHQ), 4-tert-butylpyrocatechol (In5), and 4,6-di-tert-butyl-3-sulfanyl-1,2-dihydroxybenzene (In6). In6 was the most efficient inhibitor (Ki = 11 microM at 20 degrees C in 0.015 M phosphate-citrate buffer, pH 6.0). The effects of In5 and In6 were not preceded by periods of induction of OPD oxidation products (contrary to TMHQ). Peroxidase-catalyzed oxidation of tetramethylbenzidine (TMB) was non-competitively inhibited by In6 and 3-(2-hydroxyethylthio)-4,6-di-tert-butylpyrocatechol (In4), whereas o aminophenol (OAP) acted as a mixed-type inhibitor. The effects of all three inhibitors were preceded by an induction period, during which TMB oxidation products were formed. Again, In6 was the most efficient inhibitor (Ki = 16 microM at 20 degrees C in 0.015 M phosphate-citrate buffer supplemented with 5% ethanol, pH 6.0). Judging by the characteristics of the inhibitors, taken in aggregate, it is advisable to use the pairs OPD-In5 and OPD-In6 in systems for testing the total antioxidant activity of biological fluids of humans. PMID- 14520959 TI - [Isolation of myoinositol from solutions of enzymatic biotransformation]. AB - The transferase reaction between phospholipids and inositol catalyzed by phospholipase D on phase interface in water-organic solvent systems was studied. Optimal conditions for phosphatidylinositol synthesis in water-organic solvent heterogeneous system were determined. The rapid separation of the hydrophobic components, phospholipids, from water-soluble products, alcohols, was observed in the systems with organic solvents. Displacement of myo-inositol from phosphatidylinositol by methanol, alcohol substrate, added to the reaction medium was shown in hexane-water system. Myo-inositol was isolated from the mixture of its isomers by two-stage transferase reaction catalyzed by phospholipase D. PMID- 14520960 TI - [Intracellular glucose oxidase from Penicillium funiculosum 46.1]. AB - A method for isolating extracellular glucose oxidase from the fungus Penicillium funiculosum 46.1, using ultrafiltration membranes, was developed. Two samples of the enzyme with a specific activity of 914-956 IU were obtained. The enzyme exhibited a high catalytic activity at pH above 6.0. The effective rate constant of glucose oxidase inactivation at pH 2.6 and 16 degrees C was 2.74 x 10(-6) s-1. This constant decreased significantly as pH of the medium increased (4.0-10.0). The temperature optimum for glucose oxidase-catalyzed beta-D-glucose oxidation was in the range 30-65 degrees C. At temperatures below 30 degrees C, the activation energy for beta-D-glucose oxidation was 6.42 kcal/mol; at higher temperatures, this parameter was equal to 0.61 kcal/mol. Kinetic parameters of glucose oxidase-catalyzed delta-D-glucose oxidation depended on the initial concentration of the enzyme in the solution. Glucose oxidase also catalyzed the oxidation of 2-deoxy-D-glucose, maltose, and galactose. PMID- 14520961 TI - [Comparative study of the extracellular laccases from Cerrena unicolor 059 0784 and Pleurotus oastreatus 0432]. AB - The laccases of the basidiomycetes Cerrena unicolor 059, C. unicolor 0784, and Pleurotus oastreatus 0432 were assayed comparatively. The laccases were isolated as homogenous preparations with molecular weight 55, 56, and 57 kD, respectively. The three enzymes were found to be glycoproteins. The carbohydrate moiety of the glycoproteins included mannose, galactose, and N-acetylglucosamine. The carbohydrate moiety of the laccases from C. unicolor 059, C. unicolor 0784, and P. oastreatus 0432 accounted for 17, 23, and 24%, respectively. The pH optimum of the enzymes was at 4.0, 3.75, and 5.6, respectively. Thermal stability testing of laccases at 40 degrees C revealed that the C. unicolor 0784 enzyme was characterized by the highest thermal stability (after 172-h incubation the enzyme activity was maintained at a level of 25%). The Michaelis constant (Km) values of the reactions of oxidation of pyrocatechol, hydroquinone, and potassium ferrocyanide catalyzed by the basidiomycete laccases were determined. PMID- 14520962 TI - [Participation of beta-carotene in antioxidant defense of bacterial cells]. AB - The effect of recombinant beta-carotene on the resistance of E. coli culture to menadione and paraquat was studied. The presence of beta-carotene in the E. coli cell significantly prevented an increase in the activity of superoxide dismutase induced by redox mediators but had no effect on the culture growth. The findings suggested the involvement of beta-carotene in the cell defense against oxidative stress. PMID- 14520963 TI - [Level of vitamins B in yeasts of the Saccharomyces species depending on the composition of culture media]. AB - The qualitative and quantitative composition of water-soluble vitamins B in Saccharomyces yeasts cultivated on various nutrient media was studied by high performance liquid chromatography. New strains of Saccharomyces oviformis Y-2635 and Saccharomyces vini F-5 yeasts grown in the nutrient medium with geothermal water differed in great biological value due to high intracellular concentrations of riboflavin, thiamine, nicotinic acid, and folic acid. PMID- 14520964 TI - [Phytotoxic activity of chernozem saprophytic micromycetes: specificity, sorption and stability of phytotoxins in soil]. AB - Micromycetes of the complex of typical chernozem saprotrophic fungi released phytotoxic metabolites into medium. The metabolites displayed their phytotoxic activities directly in soil. Evaluation of the toxicities, range of biological effects activities, and stabilities of phytotoxins in soil and the rates of their biodegradation allowed the species that can serve as indicators of chernozem microbial toxicosis to be selected, namely, Aspergillus clavatus, Fusarium solani, Talaromyces flavus, Penicillium rubrum, and P. funiculosum. PMID- 14520965 TI - [Isolation of Penicillium DIERCKX 1901--a producer of diketopiperazine alkaloids rokefortin and 3,12-dihydrokefortin--from permafrost]. AB - Secondary metabolites of the three strains of Penicillium aurantiogriseum, isolated from permafrost sediments, were identified. It was found that these fungi synthesized diketopiperazine alkaloids roquefortine and 3,12 dihydroroquefortine. The strain VKM FW-766 synthesized alkaloids in in the course of certain growth-related processes. When the strain was grown on a mineral medium, time courses of roquefortine and 3,12-dihydroroquefortine concentrations were characterized by biphasic curves. PMID- 14520966 TI - [Determination of the primary and fine structures of galactomannan from seeds of Gleditsia triacanthos f. intermis L]. AB - This was the first study to isolate galactomannan, a 660-kDa polysaccharide, from the seeds of Gleditsia triacanthos f. inermis L. (yield 15.4%). Its aqueous solutions were optically active ([alpha] D = +31.0 degrees) and highly viscous ([eta] = 578 ml/g). The analysis of this heteropolysaccharide using chemical, enzymatic, and chromatographic procedures, as well as IR- and 13C-NMR spectroscopy, showed that is consists of D-mannopyranose and D-galactopyranose residues (molar ratio 2.42:1). Its main chain is comprised of 1,4-beta-D mannopyranose residues, 41% of which is substituted at C-6 with single residues of alpha-D-galactopyranose. The probability of occurrence of differently substituted mannobiose units in the chain, determined experimentally, was 0.16 for the unit Man-Man, 0.50 for the units Gal(Man-Man) and (Man-Man)Gal, and 0.34 for the dissubstitued unit Gal(Man-Man)Gal. PMID- 14520967 TI - [Formation of alkaloids from Penicillium species fungi during growth on wheat kernels]. AB - The ability to produce alkaloids has been studied in 13 strains belonging to 10 species of the genus Penicillium. Most of these strains produce identical ranges of alkaloids when grown on wheat grain and synthetic Abe medium. They are roquefortine, 3,12-dihydroroquefortine, and glandicolines A and B in strain P. chrysogenum VKM F-1987; fumigaclavines A and B, festuclavine, and pyroclasine in P. commune VKM F-308, F-3491, and KBP4; agroclavine 1 and epoxyagroclavine 1 in P. fellutanum VKM F-1073; fellutanine A in P. fellutanum F-3020; roquefortine, 3,12-dihydroroquefortine, meleagrin, and glandicolines A and B in P. glandicola VKM F-743; aurantioclavine in P. nalgiovense VKM F-229; isofumigaclavines A and B, festuclavine, roquefortine, and 3,12-dihydroroquefortine in P. roquefortii VKM F-2389; roquefortine, 3,12-dihydroroquefortine, and meleagrin in P. vitale VKM F 3624; roquefortine and oxaline in P. vulpinum VKM F-256; and alpha-cyclopiazonic acid and rugulovasine B in P. viridicatum C-47. No alkaloids were found in P. rugulosum VKM F-352 grown on wheat grain. A simple method is proposed for isolating alkaloids from affected grains. PMID- 14520968 TI - [Change in the level of 1-aminochloropropane-1-carbonic acid. Activity of a protein inhibitor of polygalacturonase, intensity of formation of oligouronides in apples during ripening and treatment with haloethane derivatives and aminoethoxyvinylglycine]. AB - We studied changes in the intensity of ethylene release and accumulation of 1 aminocyclopropane-1-carboxylic acid during ripening of two apple varieties characterized by various physiological states and treated with halothane derivatives and L-alpha-(2-aminoethoxyvinyl)-glycine. We observed changes in activity of the protein polygalacturonase inhibitor in the fruit tissue and accumulation of oligouronides. The data suggest that pretreatment with the inhibitor of 1-aminocyclopropane-1-carboxylic acid synthase affects ethylene release, accumulation of 1-aminocyclopropane-1-carboxylic acid, activity of the protein polygalacturonase inhibitor, and potential intensity of oligouronide formation in apple fruits and tissues. PMID- 14520969 TI - [Structure of the photosynthetic apparatus of the bean Vicia faba L. when treated with 6-benzylaminopurine]. AB - The level of endogenous cytokinins changed with growth and development of faba bean (Vicia faba L.) leaves. Typical of juvenile leaves, amounting to 25% of the final leaf size (Smax) was a low content of these plant hormones. The level of cytokinins increased in growing leaves (50% of Smax) and decreased in the leaves that stopped growing. The content of cytokinins in senescent leaves dropped considerably. Exogenous treatment with 6-benzylaminopurine (BAP) had no effect on the structure of terminal phloem; however, it stimulated elongation of mesophyll cells; increases in the area and thickness of leaf blade, amount of photosynthetic pigments, and assimilation potential; and delayed senescence of leaves and defoliation, thereby increasing biomass of the aboveground plant part. It was inferred that BAP had a potential for induction of photosynthetic apparatus development and increase in the yield of faba bean green mass. PMID- 14520970 TI - [Aggregating ability of seed storage proteins from cereals differing in gluten quality]. AB - The effects of pH, ionic strength, and medium composition on formation of macrocomplexes of seed storage proteins from wheat, rye, and barley have been studied. It has been found that various noncovalent interactions (electrostatic and hydrophobic interactions and hydrogen bonds) are involved in protein aggregation. Their combined action depends significantly on the biochemical nature of storage proteins and on the medium. PMID- 14520971 TI - [New prospects for a method of biospecific chromatography]. AB - The polyacrylamide hydrogels with covalently immobilized ovomucoid from the duck's egg white were synthesized by radical copolymerization. These hydrogels can defend the immobilized insulin against the action of proteolytic enzymes. Biospecific interaction of the polysaccharide component of ovomucoid with lectins leads to the targeting transport of the hydrogel particles onto the small intestine wall. PMID- 14520972 TI - [Collagen fractions, obtained by water-salt extraction from animal fats]. AB - Collagen fractions have been isolated by water-salt extraction from raw materials of animal origin (various tendon types or subcutaneous tissues of cattle, or porcine skin). Collagen fractions with maximum capacity for water and fat retention were isolated with high efficiency by water-salt solutions containing 1 10% sodium chloride at temperatures below 50 degrees C. The values of the effective constant of extraction rate (min-1) at pH 6.5, 9.0, and 12.0 were equal to (2.7 +/- 0.1) x 10(-3), (6.2 +/- 0.5) x 10(-3), and (15.4 +/- 0.7) x 10(-3), respectively. The optimum conditions found made it possible to isolate collagen those proteinaceous fractions that are of practical use in food industry. PMID- 14520974 TI - Perspectives. Are uninsured caught in hospital regulatory bind? PMID- 14520973 TI - [Protein hydrolysates, obtained from crustaceans by an electrochemical method, as a basis for microbiological culture media]. AB - The utility of protein hydrolysates extracted electrochemically from crustaceans (Gammarus pulex and shrimp) as major components of microbiological nutrient media was demonstrated. Saprophytic soil bacteria of the genera Bacillus and Pseudomonas and the family Enterobactericeae displayed a good growth and typical morphology of their colonies on the experimental media in question. PMID- 14520975 TI - Perspectives. Insurers gear up to fight national health insurance. PMID- 14520976 TI - Taking the brown out of brownfields. PMID- 14520977 TI - [Recent data on Rift Valley Fever epidemiology in Senegal]. PMID- 14520979 TI - [Problems presented by obstetric deliveries in a referral center: which solutions?]. PMID- 14520978 TI - [Chloroquine resistant strains of Plasmodium falciparum in the Anopheles vector and use of antimalarials]. PMID- 14520980 TI - [Hospital essential drugs in Africa. An example of pharmacoeconomic research]. PMID- 14520982 TI - [Particularities of hepatic hydatidosis in black Africa. 32 hydatidosis cases observed in Niger Republic]. PMID- 14520981 TI - [Comparison of hepatitis B virus and hepatitis C virus seroprevalence in hepatocellular carcinoma]. PMID- 14520983 TI - [Importance of hepatic bilharziasis in liver pathologies in the internal medicine department of the National Hospital of Point "G", Bamako-Mali]. PMID- 14520984 TI - [Duodenal carcinoid tumor (a case report in Dakar)]. PMID- 14520985 TI - [Portal thrombosis in the tropical milieu. A prospective ultrasonographic study (60 cases)]. PMID- 14520986 TI - [Corneal marginal ulcer. Autoimmune diseases, new therapeutic aspects]. PMID- 14520987 TI - [Hypopharyngeal cancers in Senegal: experience of the otorhinolaryngeal clinic at the Dakar University Hospital Center 1984-1994]. PMID- 14520988 TI - [Kidney cancer in adults at the University Hospital Center of Dakar]. PMID- 14520989 TI - [Cardiac complications of hypertension: prospective study at the Principal Hospital in Dakar]. PMID- 14520990 TI - [Genetic variation in Anopheles gambiae, malaria vector. Implication in transmission]. PMID- 14520991 TI - [Malaria vectors in Senegal: a system in evolution]. PMID- 14520992 TI - [Hepatitis B and C viruses in hepatocellular carcinoma in Senegal]. PMID- 14520994 TI - Have you been rethinking your commitment to nursing? PMID- 14520993 TI - Research utilization and evidence-based practice yield favorable patient outcomes. PMID- 14520995 TI - Severe acute respiratory syndrome: is the worst of it over? PMID- 14520996 TI - "How have you used evidence-based practice to implement a practice change in your work setting?" Nurses implement evidence-based skin care practice. PMID- 14520997 TI - ONS calls for corrections to the Medicare prescription drug bill. PMID- 14520998 TI - Nursing needs your involvement. PMID- 14521000 TI - Rehabilitation nursing in a consumer-driven world. PMID- 14521001 TI - Therapeutic relationships: building blocks of rehabilitation nursing. PMID- 14521002 TI - Complementary and alternative therapy use in persons with multiple sclerosis. AB - Use of complementary and alternative therapies (CATs) is common with both the general population and with people who have multiple sclerosis (MS). The purpose of this study was to describe the reported use and perceived effectiveness of different CATs in a large sample (N = 621) of people with MS. One-third of the participants reported that they were presently using one or more CATs, and 50% had tried one or more CATs in the past. Most CATs were perceived as being helpful. People who were presently using CATs differed little from those who were not with regard to demographic- and illness-related variables. Use of CATs was positively associated with the practice of other health-promoting behaviors. The frequent use of different CATs suggests that clinicians should systematically and nonjudgmentally inquire about CAT use among their patients. PMID- 14521003 TI - Effect of a video intervention on functional recovery following hip replacement and hip fracture repair. AB - Although much research has documented the functional outcomes in hip repair surgery, few studies have identified effective interventions to improve functional outcomes. The purpose of this study was to test the use of generic and individualized videos on patients' return to preoperative levels of independent functioning, increased physical performance, improved coping ability, and enhanced perception of ability to care for self. Two rehabilitation units in separate acute care facilities were used to recruit subjects. Data were collected on 82 participants on admission to the study, on discharge from rehabilitation, 1 week after discharge, and 3 months post discharge. The results indicate that video intervention has some benefit to persons recovering from hip repair surgery. The benefits were realized in physical performance as evidenced by a significant difference in the distance walked and time walked of participants in the treatment group. PMID- 14521004 TI - Rehabilitation of a functionally limited, chronically ill older adult: a case study. AB - This case study presents MV, a 78-year-old woman who was being treated pharmacologically for several common chronic health problems and who presented with self-reported and observed limitations in her functional ability. After being screened for contraindications to exercise, MV participated in a 16-week rehabilitation program of regular exercise that included aerobic and resistance training. She maintained a high level of compliance with this rehabilitation training and showed improvements in her aerobic capacity, strength, ability to complete functional tasks, blood components, and lipid profile. She also reported decreased levels of pain and improved perceptions of functional ability. This case study indicates that older adults with chronic illness can benefit from participating in a program of regular exercise. PMID- 14521005 TI - A self-report of quality of life of patients receiving intrathecal baclofen therapy. AB - The purpose of this study was to explore through a department quality improvement tool a possible relation between quality of life (QOL), complication rates, and length of intrathecal baclofen (IB) treatment as reported by patients receiving IB therapy in a community-based rehabilitation center outpatient clinic. A second objective was to examine complication rates among the clinic's patients. No conclusions could be drawn as to the relation between QOL, various reported complications, and length of treatment. A rank order frequency of areas reported by respondents to have the greatest impact on their QOL could be extrapolated from the data collected. In addition, complication rates among the patients who responded to the survey could be reported. Surveys from 49 patients about their experiences with IB therapy were analyzed. Respondents included 30 adult and 19 pediatric patients. Thirty-six patients (73%) had used the IB pump for 1 year or more. The survey included questions about QOL, complications, and length of IB treatment. Forty-three respondents (88%) stated they felt that their QOL had improved with IB therapy. Four patients (8%) responded that they were not sure that it had, and only 2 patients (4%) said that IB had not improved their QOL. The most frequently reported positive effects on QOL were reported in the following areas: spasticity control without the sedative effect of oral medication; ease of care for caregivers; easier positioning; less pain/increased comfort; and improved patient transfers. High ratings of improvement in the patients' QOL were reported despite a reported overall complication rate of 39%. The most common complications cited were infection and catheter breakage or disconnect. The overall infection rate for respondents was 10% (5 patients of the 49 surveyed reported infection). The rate of catheter breakage or disconnect was also 10%. Despite the complications reported, 46 patients stated they would recommend baclofen treatment to others. Three patients did not respond to the question. None of the patients said they would not recommend baclofen to others. PMID- 14521006 TI - Bone up on fractures. PMID- 14521007 TI - Firewalls: cheap and easy protection. PMID- 14521008 TI - Answers to your questions about.... Business associate contracts. PMID- 14521009 TI - Malpractice. Close-up on a crisis. PMID- 14521010 TI - Career moves: think twice about academia. PMID- 14521011 TI - How to fight bundling. PMID- 14521012 TI - What to ask job candidates. PMID- 14521013 TI - Stretching the statute of limitations. PMID- 14521014 TI - Debate in newborn screening for metabolic disorders in Taiwan--a voluntary screening? Mandatory screening? Or both? PMID- 14521016 TI - Recent advances in the study of genetic and environmental risk factors of childhood leukemia. AB - In this monograph, I discuss evidence which suggests that childhood leukemia originates in utero. Furthermore, in some cases, especially those with MLL gene rearrangement (MLL+), exposure to some particular environmental toxic factors appears to increase the risk of leukemia. This exposure, together with a genetic background of genomic instability or defects in detoxifying environmental factors, may increase the risk of childhood leukemia. In light of these findings, future study should include the identification of risk factors in environmental and genetic stresses. By identifying the genetic characteristics of individuals, we can provide proper environment including maternal conditions with pregnancy. Understanding the genetic and environmental basis of childhood leukemia is a prerequisite to overcome the risks for the development of this intractable childhood disease in the future. PMID- 14521015 TI - Newborn screening: should explicit parental consent be required? AB - Newborn screening, the first population-based metabolic screening program has been universally conducted for several decades. With the advent of genetic technologies, many genetic/metabolic disorders can be detected pre symptomatically but might be untreatable. Since the benefits and risks of screening tests on newborns are not fully known, newborn screening programs confront both legal and ethical challenges. This article aims to explore the following questions: (1) Is explicit parental consent required? (2) What level of consent should be sought? and (3) Is screening in the absence of explicit consent legally defensible? This article considers: (1) the introduction of newborn screening, (2) the conditions under which it is conducted (voluntary vs. mandatory), (3) the argument over whether explicit parental consent is required, (4) the conditions under which implied consent may be assumed, and (5) the principles of informing parents. This article concludes that implied consent is not acceptable except for traditional routine screening. Healthcare professionals should provide information about the known benefits and risks of testing and recognize the parental right of refusal. Finally, since the absence of explicit parental consent to newborn screening is not legally defensible, children's advocates should lobby for legislation permitting it in the absence of explicit consent in specific circumstances. PMID- 14521017 TI - Percutaneous endoscopic gastrostomy in children: 15 cases experience. AB - For long-term tube feeding in children, percutaneous endoscopic gastrostomy (PEG) has the advantages of a short surgical time, early feeding following surgery, and lower rate of complications. From July 2000 to September 2002, we enrolled fifteen children (mean age: 8.2 years old) who underwent PEG placement for long term nutritional support. Their underlying diseases included mucopolysaccharidosis (MPS) type II severe form, mitochondrial disease, Ehlers Danlos syndrome associated with Robin sequence, spinal muscular atrophy (SMA) type II, nesidioblastosis, neurofibromatosis and other neurological disorders. We assessed the complications and outcome in these patients after PEG placement. There were no difficulties in PEG tube-feeding after procedure. One patient had a wound infection at the insertion site which required parenteral antibiotic therapy. Symptomatic gastroesophageal reflux (GER) occurred in two patients and was controlled with medication. One patient developed a gastrocutaneous fistula, requiring surgical removal of the PEG tube. One patient underwent laparoscopic gastrojejunostomy and Nissen fundoplication for persistent vomiting. Two patients with mitochondrial disease expired. The other eleven devices have continued to function on follow-up. Placement of a PEG is a simple, feasible procedure for children with swallowing difficulty who require long-term nutritional support. Although complications may sometimes occur, in our experience many can be managed conservatively. PMID- 14521018 TI - Maternal intention and actual behavior in infant feeding at one month postpartum. AB - Five-hundred and ninety-one mothers completed a follow-up questionnaire at one month postpartum. The total breastfeeding rate declined from 83.4% before discharge to 50.8% at one month. One-hundred and fifty-three (25.9%) mothers exclusively breastfed, 147 (24.9%) mothers mixed fed, and 291 (49.2%) mothers formula fed their babies at that time. Among the mothers who initially intended to exclusively breastfeed, 39.3% changed to formula feeding, while 7.1% of the mothers who intended to mix feeding and 4.8% of the mothers who intended to formula feed changed to exclusive breastfeeding at one month postpartum. Nine percent of the breastfeeding mothers weaned within the 1st week, 9.2% weaned between the 1st and 2nd week, 8.4% weaned between the 2nd and 3rd week, while another 14.8% weaned between the 3rd and 4th week. Milk insufficiency, maternal tiredness, not knowing whether the infant had enough to eat and breast problems were the four major reasons for changing feeding method from breastfeeding to formula feeding. Positive breastfeeding attitude, non-employment and husband's approval of breastfeeding was negatively associated with changing feeding behavior from breastfeeding to formula feeding within one month postpartum. In conclusion, successful breastfeeding not only depends on factors related to the mother, but also on her environment. Education regarding breastfeeding should be provided to the entire family including the mother and father. The workplace needs to be more accommodating to breastfeeding mothers, so that they can continue breastfeeding after returning to work. PMID- 14521019 TI - Thyroid function in children with newly diagnosed type 1 diabetes mellitus. AB - Evaluation of thyroid hormone indices was performed in 138 children with newly diagnosed type 1 diabetes, with their siblings serving as controls. The DKA group consisted of 76 children who had diabetic ketoacidosis (DKA) at initial diagnosis. The non-DKA group consisted of 62 children and the control group of 35. The thyroid function tests of the patients were measured within 3 days of the initial diagnosis of diabetes and at least one follow-up test one month to two years after adequate treatment of diabetes. The DKA group had significantly lower levels of T3, T4, free T4 and FTI than did the other two groups (p < 0.0001, p < 0.0001, p < 0.0001, and p < 0.0001, respectively). T3 concentration was lower in non-DKA subjects than in controls (p = 0.0003), but the two groups did not significantly differ in terms of T4, free T4, and FTI. The TSH level did not differ among the three groups. We conclude that DKA changes thyroid function measurements. In the absence of true thyroid disease, abnormal thyroid function tests are reversible after institution of good diabetic control. We suggest that thyroid function tests should be restricted to those patients suspected of having thyroid disorders at the initial diagnosis of type 1 diabetes. PMID- 14521021 TI - Congenital myotonic dystrophy: report of one case. AB - We report a laboratory-confirmed pediatric case of congenital myotonic dystrophy (CMD) in Taiwan. A 5-year-old girl presented in our neurological clinic with the chief complaints of developmental delay and slowness in learning. Clinical examination, intelligence tests, and developmental examinations revealed myopathic facial weakness, passivity and delayed cognitive development. Past history review showed neonatal hypotonia, feeding difficulty with cyanosis, and poor activity. Neonatal studies only revealed subgaleal hematoma and subarachnoid hemorrhage with mild degree right side intraventricular hemorrhage. Current neurological examination disclosed percussion myotonia and myotonia as poor muscle relaxation after forceful handgrip. CMD was confirmed by electromyography study and DNA analysis. The patient's mother, though minimally affected, had similar laboratory results. CMD affects children in both motor and cognitive function; the diagnosis should be entertained in developmentally delayed children with subtle neurological signs, even without strong family history. PMID- 14521020 TI - Effects of maternal nicotine exposure on branching morphogenesis of mouse fetal lung: in vivo and in vitro studies. AB - Epidemiological evidence suggests that premature infants born to mothers who smoke have a lower incidence of neonatal respiratory distress syndrome. The mechanism has been proposed to be due to increased lung maturity. This in vivo study investigated the effect of maternal nicotine on lung development by evaluating the airway branching morphogenesis (ABM) in mice fetuses. Nicotine (0, 2 and 3 mg/kg/day) was administered intraperitoneally to pregnant mice from gestation day 9 to day 12 (4 days). ABM was determined on day 13 by photomicrographic analysis. The results revealed a significant reduction in ABM in the higher dose nicotine group. The mean number of airway branches was 3.7 +/- 0.1/lobe for the 3 mg/kg/day group, which was smaller than 4.6 +/- 0.2/lobe for the 2 mg/kg/day nicotine group, and 4.4 +/- 0.1/lobe for the control group (F = 9.4, p < 0.001). The mean number of buds was significantly smaller in both the 2 mg/kg/day group and the 3 mg/kg/day group (8.7 +/- 0.5/lobe, 9.0 +/- 0.4/lobe vs. 12.3 +/- 0.4/lobe in the control group, F = 20.3, p < 0.001). For the in vitro study, fetal lung lobes were isolated at the 12th gestation day. The lung explants were cultured in nicotine (0, 30, 60 ng/ml) for 48 hours; there were no differences in all the groups. The results do not support the hypothesis that nicotine stimulates fetal lung ABM either in vivo or in vitro. PMID- 14521022 TI - Systemic lupus erythematosus with presentation as vertigo and vertical nystagmus: report of one case. AB - An 11-year-old boy suffered from fever, headache, severe vertigo and unsteady gait. Physical examination showed bilateral vertical nystagmus, mild corneal reflex delay of the right eye and asymmetric facial expression. Laboratory data showed leukopenia, high ESR and normal CSF study. Brain CT showed diffuse brain edema. Electronystagmography showed upbeat nystagmus and central vertigo. EEG revealed diffuse slow wave and mild to moderate cortical dysfunction. MRI of the head showed focal abnormal signal intensity at the ventral portion of the medulla oblongata on both sides. Under suspicion of enteroviral encephalitis, mannitol and IVIG were given. The virological profiles were negative, ANA 1:640 nucleolar type, low complements and proteinuria. Anti-ds DNA was elevated and anti ribosomal-P antibodies were positive. Under impression of SLE with CNS involvement, betamethasone was given. Fever, nystagmus and ataxia subsided gradually. Steroid was tapered and imuran was added. The following laboratory data were normal. In his past history, the patient was diagnosed Kikuchi disease. The manifestations of SLE were rare initial presentations as vertigo or vertical nystagmus. We present a case with review of literature and conclusion that physicians should keep in mind the possibility of SLE if patients present with unspecific neurological symptoms and concomitant systemic symptoms. PMID- 14521023 TI - Smith-Magenis syndrome: report of one case. AB - Smith-Magenis syndrome (SMS) is a multiple congenital anomalies/mental retardation syndrome characterized by an interstitial deletion on the short arm of chromosome 17 involving the band p11.2. A 3-year-old girl was referred for evaluation of moderate psychomotor retardation and several behavioral problems including self-injuring behavior, hyperactivity, and sleep disturbance. Visual and hearing impairment, and brain abnormalities including ventriculomegaly and hypoplastic right transverse sinus, were detected. Routine cytogenetic study showed an apparent mosaicism of an interstitial deletion over the chromosomal region 17p11.2 in 65% metaphases. However, the microdeletion was found in all metaphases by fluorescence in situ hybridization study with the probe D17S29 (for Smith-Magenis critical region on 17p11.2). Her sleep disorder improved after the use of melatonin. PMID- 14521024 TI - Supraventricular tachycardia after fenoterol inhalation: report of two cases. AB - Supraventricular tachycardia (SVT) following fenoterol inhalation in metered-dose inhaler (MDI) has never been reported. We report two cases of SVT after fenoterol inhalation in MDI. Case one was a 4-year-old boy who had asthma since early childhood. Paroxysmal supraventricular tachycardia (PSVT) was found after fenoterol inhalation (MDI), which returned to normal sinus rhythm following adenosine injection. The other one was a 9-year-old male who also had asthma since early childhood. He suffered from attacks of PSVT four times after fenoterol inhalation within one year. After verapamil injection and vagal maneuvers, PSVT was converted to normal sinus rhythm. There were no other episodes of SVT after discontinuing usage of fenoterol inhalation for 2 years in the follow-up. We report these two cases to remind pediatricians that cardiac arrhythmias should be evaluated following fenoterol inhalation (MDI). PMID- 14521025 TI - Vascular ring due to double aortic arch with atretic left arch and left ligamentum arteriosum: report of one case. AB - A 1-day-old female infant presented with vomiting immediately after feeding and shortness of breath after birth. Esophagography revealed external compression of the esophagus. Echocardiography showed a right aortic arch with mirror image branching of brachiocephalic vessels without intracardiac anomalies. Left ventriculography confirmed the echocardiography findings and revealed a tenting of the proximal part of the left subclavian artery and a blind pouch of Kommerell diverticulum at the descending aorta. The patient underwent surgery, which revealed a complete vascular ring compressing the trachea and the esophagus. The ring was formed by a right aortic arch, atretic left arch and left ligamentum arteriosum. Following surgical division of the ligamentum arteriosum and the atretic left arch, the symptoms subsequently improved. PMID- 14521026 TI - Living-related liver transplantation for methylmalonic acidemia: report of one case. AB - Methylmalonic acidemia (MMA) is an autosomal recessive disease of branched chain amino acid metabolism. The disease can be classified as vitamin B12-responsive or nonresponsive type. For the nonresponsive type, there is no curative treatment, and its prognosis is usually poor. We report one case that had vitamin B12 nonresponsive type MMA diagnosed at 3 days of age. The patient received liver transplantation at the age of 11 months. Continuous venovenous hemodialysis was performed immediately pre- and post-transplantation. The attack frequency of metabolic acidosis episodes decreased and the general condition of this patient improved after liver transplantation. So, although not curative, liver transplantation is an effective management for MMA. PMID- 14521027 TI - Kawasaki disease presenting with hepatitis and prolonged fever: report of one case. AB - Kawasaki disease predominantly affects children younger than 5 years. Coronary artery aneurysms were found in around 20% of untreated patients. We report on a case of a 10-year-old boy who had atypical presentation of Kawasaki disease with significant hepatobiliary dysfunction, including hepatomegaly and jaundice, and persistent fever. He did not have conjunctivitis until the 8th day of fever, and periungual desquamation and strawberry tongue until the 13th day of fever when Kawasaki disease was diagnosed. Echocardiography revealed multiple coronary artery aneurysms. Such atypical clinical pictures of Kawasaki disease may cause delay in the treatment, and the fatality rate can be increased. So, a persistent fever with jaundice should evoke the differential diagnosis of Kawasaki disease. PMID- 14521028 TI - USP publishes enforceable chapter on sterile compounding. PMID- 14521029 TI - Staffing shortages plague nation's pharmacy schools. PMID- 14521030 TI - Medicare reform could harm Medicaid, 340B program. PMID- 14521031 TI - Clinic for identifying and addressing polypharmacy. PMID- 14521032 TI - Medical education and communication companies: how ideal a fit for pharmacists? PMID- 14521033 TI - Computerized prescriber order entry: power, not panacea. PMID- 14521034 TI - Bivalirudin in percutaneous coronary intervention. AB - The chemistry and pharmacology, pharmacokinetics, pharmacodynamics, adverse effects, drug interactions, dosing and administration, and pharmacoeconomics of bivalirudin are reviewed; clinical trials of bivalirudin's application in percutaneous coronary intervention (PCI) are also discussed. Bivalirudin is a direct thrombin inhibitor approved for use in PCI. It reversibly binds to thrombin's catalytic site and substrate recognition site and blocks both circulating and fibrin-bound thrombin. Peak concentrations occur in less than 5 minutes after bolus-dose administration, and its half-life is approximately 25 minutes. It is primarily eliminated renally, and dosage reduction may be required in patients with severe renal dysfunction. Two clinical trials have demonstrated that bivalirudin is at least as effective as unfractionated heparin (UFH) in preventing ischemic complications in PCI. Other trials have shown that bivalirudin has beneficial ischemic and hemorrhagic outcomes in a more modern PCI setting (i.e., intracoronary stent placement, clopidogrel, and glycoprotein IIb/IIIa-receptor inhibitors). Bivalirudin combined with provisional glycoprotein IIb/IIIa inhibitors was noninferior to UFH with planned glycoprotein IIb/IIIa inhibitors and superior to UFH alone with respect to ischemic and hemorrhagic endopoints in PCI. Major bleeding with bivalirudin has occurred in approximately 3% of patients in clinical trials, and it is not known to have any interactions with the cytochrome P-450 isoenzyme system. The acquisition cost of bivalirudin in one study was less than the combination of UFH and glycoprotein IIb/IIIa inhibitors. Bivalirudin combined with provisional glycoprotein IIb/IIIa inhibitors appears to be an acceptable alternative to the standard of care and is superior to UFH alone in PCI. PMID- 14521035 TI - Recovery of phenytoin from feeding formulas and protein mixtures. AB - The recovery of phenytoin from mixtures containing different phenytoin formulations and protein mixtures was studied. Three phenytoin solutions (40 mg/microL) were prepared, each in triplicate, from phenytoin tablets, phenytoin suspension, and bulk phenytoin powder. These solutions were mixed with equivalent volumes of two commercially available feeding formulas (Replete and Ultracal) and two isolated protein mixtures (casein protein mixture and why protein isolates mixture) and placed in ultrafiltration tubes. The mixtures were centrifuged, and phenytoin recovery was determined by using high-performance liquid chromatography. Control data were also obtained before and after the experiment. There was no difference in phenytoin recovery when comparing phenytoin tablets versus phenytoin suspension in any of the protein media. There was a significant difference in phenytoin recovery when comparing the standard phenytoin solution mixed with Replete (32.51%) versus Ultracal (37.71%). There was also a significant difference in recovery when comparing the standard solution mixed with the calcium caseinate mixture (48.41%) versus the whey protein isolates mixture (82.01%). While the difference in recovery between Replete and Ultracal was expected, the significantly higher recovery of phenytoin from the whey protein mixture versus the calcium caseinate mixture indicated a much lower binding affinity between phenytoin and whey protein than with phenytoin and casein. The recovery of unbound phenytoin from feeding formulas and solutions of protein isolates did not differ with phenytoin formulations. Ultracal had a lower level of binding to phenytoin than Replete; whey protein had a lower level of binding than casein. PMID- 14521036 TI - Using medium-fill simulation to establish a benchmark microbiological contamination rate for low-risk-level compounding. AB - A benchmark contamination rate for prefilled syringe compounding was determined by using a medium-fill-simulation method. One thousand thirty-five 1-mL tuberculin syringes were aseptically filled with 0.9 mL of sterile soybean-casein digest medium and capped. These syringes were placed into clear nonsterile plastic bags and incubated at 35 degrees C for seven days, then inspected for cloudiness or colony formation indicative of bacterial growth. The operation was performed by two certified pharmacy technicians in an ISO Class 5 vertical airflow biological safety cabinet in a typical inpatient pharmacy compounding area over two days. The technicians wore protective garments and gloves and followed standard procedures of preparing sterile injections. The adequacy of the methodology was verified by using four syringes that contained deliberately contaminated medium and incubating them along with 15 aseptically prepared syringes at 35 degrees C for seven days. Colony formation in the four syringes containing contaminated medium was directly observed and differentiated from the control syringes, which confirms the validity of the method. No bacterial growth was detected in any of the 1035 medium-filled syringes studied. Therefore, the contamination rate for aseptic compounding operation was less than 0.1%. Medium fill-simulation testing of 1035 prefilled tuberculin syringes yielded no contamination. A contamination rate of less than 0.1% should be achievable and expected for this type of low-risk pharmacy preparation after pharmacies validate their own compounding operation. PMID- 14521037 TI - A pharmacist-managed clinic for treatment of latent tuberculosis infection in health care workers. AB - A pharmacist-managed clinic for treating latent tuberculosis infection (LTBI) in health care workers (HCWs) is described. The hospital-based clinic was begun in 1993 after a review showed that only 0.8% of HCWs at the institution who were prescribed isoniazid for LTBI completed it. The goals of the clinic are to improve HCWs' adherence to LTBI treatment, enhance outcomes, and minimize adverse drug reactions. The clinic is staffed by a pharmacist, a medical resident, and a nurse. If an employee is diagnosed with LTBI, he or she is referred to the clinic, where the pharmacist conducts an initial assessment and provides information on medication available for treatment. The worker is then scheduled for monthly follow-up visits with the pharmacist for the first three months of LTBI therapy, during which the pharmacist evaluates treatment adherence and potential adverse effects. Thereafter, the pharmacist interviews the patient monthly by telephone until therapy is complete. HCWs are allowed only a one-month supply of medication with each refill. From June 1993 through June 1997, of 131 employees who started treatment, 122 (93%) completed it. From June 1997 through December 2001, annual therapy completion rates ranged from 90% to 100%. No cases of active tuberculosis have been reported in treated patients, and only nine adverse drug reactions were reported for the 1997-2001 period. A pharmacist managed clinic substantially improved rates of treatment completion among HCWs with LTBI. PMID- 14521038 TI - Survey of state requirements for unit-of-use packaging. PMID- 14521039 TI - Developing and assessing requirements for clinical decision support. PMID- 14521040 TI - Pharmacist interventions after implementation of computerized prescriber order entry. PMID- 14521041 TI - Thrombolytic administration in the management of subarachnoid hemorrhage. PMID- 14521042 TI - Medical education and communication companies: career options for pharmacists. PMID- 14521043 TI - Am I still a pharmacist? PMID- 14521044 TI - Scientific writing: a career for pharmacists. PMID- 14521045 TI - Continuing medical education: a view from the inside. PMID- 14521046 TI - Doing what needs to be done in pharmacy practice leadership: a message for residents. PMID- 14521047 TI - Neural networks: from science fiction to pharmacy. PMID- 14521048 TI - New electronic device for monitoring medication compliance. PMID- 14521049 TI - Genes, smoking, and treatment response. PMID- 14521050 TI - Appropriateness of statistical analysis. PMID- 14521051 TI - [The role of the sarcocyst surface apparatus in utilizing the host cell muscle structures]. AB - The participation of the sarcocyst surface apparatus (SSA) of two sarcosporidian species, Sarcocystis muris and S. ovifelis (Coccidia, Sporozoa, Apicomplexa), in degradation of disrupted host cell substances was investigated. After degradation, these substances are transported through the membrane of the SSA to the sarcocyst ground substance (GS), but this process cannot be regarded as endocytosis. At first, the transported substances were found in SSA pits in the form of fibrillar structures. Later on, these were seen as twisted up granules. In some cases, such granules restore their fibrillar shape, penetrate through the SSA membrane and appear in the sarcocyst GS. In other cases, the small granules may be released from SSA pits directly to the sarcocyst GS. Besides, two SSA primembrane layers were seen to disappear during the transportation of host cell substances. In addition, multimembrane structures (membranous whorls) were first demonstrated between the plasmalemma and inner membrane complex of the zoite pellicle. Multimembrane structures were found, in addition, in the zoite cytoplasm in connection with micronemes. These structures resembling chloroplast granae of thylakoids may presumably fill the gap in membrane pool of the SSA contributing to its renewal. PMID- 14521052 TI - [Morpho-functional analysis of the steroid-producing cells in pink salmon fry gonads Oncorhynhus gorbuscha (Walbaum) in normal condition and after estradiol treatment]. AB - Localization and peculiarities of steroid-producing cells of gorbuscha fry gonads were studied in normal conditions and after experimental conditions. In females steroid-producing cells are found also in the follicle tunica and between interstitial cells, greatly differing in ultrastructure in these two localities. In males, these cells are located only in the testicular tunica, being homogeneous in ultrastructure. Chronical hormonal treatment of the fry with oestradiol-dipropionate induced in males, contrary to females, a considerable increase in the number and functional activity of steroid-producing cells and the appearance of these cells in the stroma of testis also. In females only a mass appearance of lipid-like droplets in the cytoplasm of steroid-producing cells is observed, which is also characteristic of males. The revealed morpho-functional peculiarities of steroid-producing cells are discussed in terms of their presumed important role in regulation of sex differentiation in fishes. PMID- 14521053 TI - [Structural and functional analysis of glucose adsorption at high maltose concentrations in the rat small intestine in vivo]. AB - To elucidate the mechanism of glucose absorption at high substrate concentrations, we studied structural and ultrastructural peculiarities of enterocytes arranged at different levels along the intestinal villus. The preparations were obtained from an isolated segment of the rat small intestine after its perfusion with maltose solutions with both low (25 mM) and high (100 mM) concentrations, respectively. Under conditions of chronic experiment at high substrate concentration, an enlargement of intercellular clefts, indicating glucose absorption, occurred in deeper areas of the villus. Besides, also in chronic experiment, we studied kinetics of maltose hydrolysis and derived glucose absorption in the isolated segment of the rat small intestine after its perfusion with maltose at superhigh (up to 200 mM) initial concentrations. Based on these data, a conclusion is made that active transport is the main mechanism of absorption of glucose derived from maltose hydrolysis, operating both at low disaccharide concentrations, and in the range of its superhigh (up to 200 mM) concentrations. PMID- 14521054 TI - [STAT1 and STAT3 activation by oxidative stress in A431 cells involves Src dependent EGF receptor transactivation]. AB - Different cellular signal transduction cascades are affected by environmental stressors (UV-radiation, gamma-irradiation, hyperosmotic conditions, oxidants). In this study, we examined oxidative stress-evoked signal transduction pathways leading to activation of STATs in A431 carcinoma cells. Oxidative stress, initiated by addition of H2O2 (1-2 mM) to A431 cells, activates STAT3 and, to a lesser extent, STAT1 in dose- and time-dependent manner. Maximum phosphorylation levels were observed after a 2 minutes stimulation at 1-2 mM H2O2. Phosphorylation was blocked by AG1478, a pharmacological inhibitor of the epidermal growth factor receptor tyrosine kinase, implicating intrinsic EGF receptor tyrosine kinase in this process. Consistent with this observation, H2O2 stimulated EGFR tyrosine phosphorylation was abolished by specific Src kinase family inhibitor CGP77675, implicating Src in H2O2-induced EGFR activation. An essential role for Src and JAK2 in STATs activation was suggested by three findings. 1. Src kinase family inhibitor CGP77675 blocked STAT3 and STAT1 activation by H2O2 in a concentration-dependent manner. 2. In Src-/-fibroblasts, activation of both STAT3 and STAT1 by H2O2 was significantly attenuated. 3. Inhibiting JAK2 activity with the specific inhibitor AG490 reduced the level of H2O2-induced STAT3 phosphorylation, but not STAT1 in A431 cells. These data show essential roles for Src and JAK2 inactivation of STAT3. In contrast, H2O2 mediated activation of STAT1 requires only Src kinase activity. Herein, we postulate also that H2O2-induced STAT activation in carcinoma cells involves Src dependent EGFR transactivation. PMID- 14521055 TI - [Dependence of EGF receptor and STAT factor activation on redox of A431 cells]. AB - Reactive oxygen species (ROS) were established to play an important role in cellular signaling as second messengers by integrating different pathways. Recently, we showed that EGF initiated a rapid tyrosine phosphorylation of both EGF-receptor and STAT factors with simultaneous increase in the intracellular ROS level. Now, we have investigated the effect of intracellular red-ox state on EGF- and H2O2-induced activation of EGF receptor, STAT1 and STAT3. We demonstrated that the pretreatment of A431 cells with antioxidant N-acetyl-L-cysteine (NAC) partly reduced the level of EGF-induced phosphorylation of proteins under investigation. Besides, H2O2-induced activation of EGF receptor, and STAT factors was fully prevented by NAC pretreatment. The inhibition of ROS generation by DPI declined EGF-dependent activation of EGF receptor and STAT factors to basal level. Our results demonstrate the essential role of cellular red-ox status in the modulation of EGF-mediated activation of receptor and STAT factors. We have postulated that EGF-induced ROS generation is a very important initial event promoting physiological activation of EGF receptor and subsequent STAT factor activation. PMID- 14521057 TI - [MAP-kinase cascade analysis in transformed cell with different abilities to make G1/S block under serum starvation]. AB - We have studied the ability of ERK1,2, JNK1,2 and p38 kinases to be regulated after serum deprivation in E1A + E1B-19 kDa- and E1A + E1A + c-Ha-ras-transformed rat embryo fibroblasts. It was demonstrated that oncogene transformation resulted in an increase of total kinase content independently of the type of complementing oncogene. However, for ERK1,2 kinases phosphorylation was found to depend on the type of complementing oncogene. Besides, unusual biphasic character for ERK1,2 kinases phosphorylation was checked in control fibroblasts REF52 and in transformed E1A + E1B-19 kDa cells, which undergo G1/S arrest after a 24 h serum starvation. According to the immunoblotting data, phosphorylated forms of ERK1,2 kinases are not detected after 15-30 min of serum deprivation, but their content is restored up to the control level within several hours. At the same time, the level of ERK1,2 phosphorylation in E1A + c-Ha-ras cells did not change after serum withdrawal. Besides, serum deprivation did not lead to significant changes in the level of phosphorylation of both type stress kinases--JNK2 and p38 in all types of studied cells. We discuss possible mechanisms of biphasic alteration in ERK1,2 phosphorylation level under condition of serum deprivation of REF52 cells and E1A + E1B-19 kDa-transformed fibroblasts, able to be arrested in G1 phase. PMID- 14521056 TI - [EGF regulation of specific endoribonuclease activity of 26S proteasomes from A431 cells: a potential role of proteasomes in control of mRNA stability]. AB - For the first time it has been shown that RNase activity is induced under the influence of EGF on epidermoid carcinoma cell line A431. Proteasomes from EGF treated A431 cells destabilize the 3'-untranslated regions of non-muscle beta actin mRNA, creating a specific cleavage pattern. In addition, these particles have been shown to specifically cleave Alu-containing informational RNA. The enzymatic activity under study has been shown to be dependent on phosphorylation of proteasomal subunits and specifically and selectively regulated by Ca and Mg ions. Proteasome involvement in the coordinated control of stability of specific messenger RNA molecules is suggested. The endoribonuclease activity of 26S proteasomes can constitute a link between EGF signaling pathways and RNA stability. PMID- 14521058 TI - [Water and ion balance in rat thymocytes under apoptosis induced with dexamethasone or etoposide. Ion-osmotic model of cell volume decrease]. AB - Cell ion and water balance was studied with respect to analysis of the osmotic model of apoptotic volume decrease (AVD) in rat thymocytes under dexamethasone (1 microM, 4-6 h) or etoposide (50 microM, 5 h) treatment. Intracellular water content was determined by measurement of cell buoyant density in continuous Percoll gradient, while intracellular potassium and sodium contents were determined by flame emission analysis. Apoptosis was verified by an increase in cell buoyant density, fluorescence of cells stained with Acridine orange and Ethidium bromide (flow cytometry), by changes in the cell cycle and the appearance of sub-diploid peak in the DNA histogram (flow cytometry), and by a decrease in cell size examined with light microscope. A separate fraction of dense cells with reduced size was found to appear after dexamethasone or etoposide treatment. This fraction was considered as apoptotic. An increase in buoyant density of apoptotic cells corresponded to a decrease in cell water content. In apoptotic cells vs. cells with normal buoyant density, the intracellular potassium content was lower, but sodium content was higher. The sum of potassium and sodium contents was lower in apoptotic cells. Taken into account the loss of anions, associated with the loss of cations, the bulk decrease in ions content has been sufficient to be accounted for cell volume decrease on the basis of the ion-osmotic model. PMID- 14521059 TI - [Mating types in the ciliate Dileptus anser. Genetical instability in the mating type system]. AB - In F1 and F2 from a cross between two clones of Dileptus anser isolated from natural sources (MT 1 x x MT III; MT = mating type), along with "normal" clones, many clones were observed demonstrating abnormal phenotype with respect to the MT character. Irregular features of the latter were as follows: a) a delay in maturation; b) temporary reversion to immature or adolescent state, which means instability of maturity state; c) expression of MT I and MT III, rather than MT II as in properly matured clones; d) changes in MT (i.e., MT instability); e) appearance of totally unexpected MTs in terms of the scheme of genetic control of MTs in D. anser previously suggested by Afon'kin and Yudin (1987)--e.g., of all three MTs in F1 from the initial (analysing!) cross. Amazingly, these abnormal D. anser clones closely resembled some selfer-clones of Tetrahymena pigmentosa, previously reported as an example of genetic instability in the ciliate MT system (Simon, 1980; Simon, Orias, 1987). PMID- 14521060 TI - [Chromosome mutations in marshland and dry valley populations of Abies sibirica Ledeb]. AB - Results of chromosome mutation investigation in Abies sibirica populations growing in fen and dry valley in Western Siberia are presented. The frequency of chromosome aberrations and mitotic irregularities is higher in dry valley population of the fir, than in its fen population. However, the spectrum of abnormalities registered in metaphase and ana-telophase cells of the fir from fen is broader, compared to that from dry valley. These features may be due to the fir genome reorganization in different environmental conditions, and indicate the plant reaction to stress factors. PMID- 14521061 TI - [Intensive drying of grids after lead citrate staining lowers precipitate contamination on the surface of ultrathin sections]. AB - Two techniques are proposed to minimize precipitate contamination of ultrathin sections after lead citrate staining. According to the first one, stained and washed grids are placed, sections downward, on a stack of filter papers. The second one involves a consecutive washing of grids with distilled water, 96% ethanol, and n-hexane. Both techniques are equally efficient; the former being simpler in processing, while the latter is superior in the reproducibility of results. PMID- 14521062 TI - [Pangenome]. AB - A conception of general interaction between genetic information of different organisms is formulated. A new concept "Pangenom" is proposed for description of the total sum of genes of living organisms and viruses. Features of the Pangenom structure and processes controlling its saving and variability are discussed. It is demonstrated that this conception may be used in medicine for the aims of prognosis of epidemic situation, in particular, of newly spreading pathogen development of new methods of prophylaxis and diagnostics of oncological diseases, and also of the evaluation of possible consequences of molecular genetics and gene engineering interventions. PMID- 14521063 TI - Risk assessment for venous thromboembolism in acute medical illness. PMID- 14521064 TI - Guidelines are not directives. PMID- 14521065 TI - Clinical management and algorithms in osteoporosis. AB - The aim of management in osteoporosis is to reduce the risk of fractures. Case finding strategies are currently recommended to identify subjects at high risk of future fracture who will benefit most from therapy. This article reviews these approaches and discusses the current and future treatment options that are available for patients with osteoporosis. PMID- 14521067 TI - DXA scanning and its interpretation in osteoporosis. AB - The recent growth in the clinical use of bone densitometry began with the introduction of the first dual X-ray absorptiometry (DXA) scanning systems in the late 1980s. Today, scans to measure bone mineral density are seen as having an essential role in the evaluation of patients at risk of osteoporosis. PMID- 14521066 TI - The epidemiology and scale of the problem. AB - Osteoporosis is an important health problem because of its association with fragility fractures. This article reviews the epidemiology of osteoporosis including its occurrence, risk factors and health impact. PMID- 14521068 TI - Stapled haemorrhoidectomy: a novel procedure. AB - Stapled haemorrhoidectomy is an effective procedure for symptomatic haemorrhoids with mucosal prolapse. The advantages over the conventional surgical technique include reduced postoperative pain, shorter hospital stay and earlier return to work. PMID- 14521069 TI - Managing vascular impairment following orthopaedic injury. AB - Orthopaedic trauma is associated with vascular impairment in approximately 1% of cases, complicating management of the injury, particularly to the shoulder, elbow, pelvis and knee. Doctors treating trauma would be expected to detect and manage several such patients a year. PMID- 14521070 TI - Helicobacter pylori: a clinician's view. AB - Helicobacter pylori infection has been linked with a number of gastrointestinal diseases, such as peptic ulcer disease, gastric mucosa-associated lymphoid tissue lymphoma and gastric cancer. This article reviews some of the evidence for these associations, and discusses the latest recommended indications for eradication therapy. PMID- 14521071 TI - Helicobacter pylori: a microbiologist's view. AB - Helicobacter pylori infection is known to cause a number of gastrointestinal diseases. From the standpoint of a microbiologist there are several important facets to this organism, which include the nature of the organism, it's diagnosis and the effectiveness of eradication therapy. PMID- 14521072 TI - Modern dressings: healing surgical wounds by secondary intention. AB - There is a lack of robust research evidence on many aspects of wound care. The inaugural Professional Select Committee on Wound Care, held at the Royal College of Surgeons, explored the relative value of modern and traditional dressings in treating wounds healing by secondary intention. This article outlines its main findings. PMID- 14521074 TI - A survey of the safety of doctors driving home after a night 'on call'. AB - The 'on-call' system for doctors within the health service maintains a continuity of care. However, allowing doctors to drive home after a night on call may pose a public hazard. The aim of this survey was to assess doctors' opinions regarding the safety of driving without adequate sleep. PMID- 14521073 TI - The eye in gastrointestinal disease. PMID- 14521075 TI - Dancing eyes. PMID- 14521076 TI - Euglycaemic diabetic ketoacidosis. PMID- 14521077 TI - The NHS and leadership. PMID- 14521078 TI - The NHS and leadership. PMID- 14521079 TI - The NHS and leadership. PMID- 14521080 TI - The NHS and leadership. PMID- 14521081 TI - The NHS and leadership. PMID- 14521082 TI - Anaesthesia for coronary artery bypass: should it differ off-pump and on-pump? PMID- 14521083 TI - [Basic types and vital forms of human peripheral and PHA-stimulated lymphocytes studied in vitro]. AB - Natural diversity in peripheral and PHA-stimulated lymphocytes seen in the same donors was studied using digitized streak photo of living cells in observational camera. Cells were monitored for 5-8 h at the superior limit of optical resolution by means of phase-contrast microscopy. Intact lymphocytes were observed in autological blood plasma, and PHA-stimulated lymphocytes were examined in self-conditioned centrifuged growth medium. The majority of intact cells were small- and middle-sized floating lymphocytes with microvilli, and middle-sized caudate lymphocytes capable of stick-slip motion. The lesser part consisted of "spread-eagle" or movable forms of both large granular lymphocytes and middle-sized lymphocytes of several types: narrow-plasm lymphocytes with lamellipodia, wide-plasm lymphocytes without cytoplasmic processes, lymphocytes with single pseudopodia, and lymphocytes with single lobopodia of complex shape. On the contrary, the minor fraction of PHA-stimulated lymphocytes of 3 day old cultures contained floating cells with microvilli or floating cells with microvilli and two pseudopodia, whereas the majority of these lymphocytes were spread-eagle or movable forms of cells of different type. These substrate adhesive PHA-stimulated lymphocytes had well defined apical and basal cell surfaces, but upon mechanical stress are easily pinched off to become ball shaped. At least 6 different cell types were distinguished among substrate adhesive PHA-stimulated lymphocytes, with more than half of these being heavily vacuolated spheroid lymphocytes prone to forming cell clusters. The rest PHA stimulated lymphocytes were represented by signet-ring lymphocytes with dark or light cytoplasm, narrow-plasm lymphocytes with large prolonged nuclei and lamellipodia, lymphocytes with single lobopodia, and lymphocytes with single spiral structures in the cytoplasm. The spiral structure is 10-11 microns in length and 0.5-0.7 micron in width, being presumably a mitochondrion or a group of butt-joined mitochondria. Since some of the caudate middle-sized lymphocytes also contain this structure, these may be regarded as putative precursors of respective type of PHA-stimulated lymphocytes. Under the conditions of observation, interphase nuclei of all live PHA-stimulated lymphocytes were seen to contain numerous globular or fiber structures of condensed chromatin made of 0.3-0.8 micron beads. These beads are doubtless interphase chromomeres. PMID- 14521084 TI - [Endocrine cells of mucosal epithelium in the distal gut of the grass frog Rana temporaria]. AB - The epithelium of the distal part of Rana temporaria intestine has been studied by light and electron microscopy. It has been shown that the number of agrent affined endocrinocytes in the mucosa of the frog intestinal epithelium decreases gradually from the small bowel to the colon, and then sharply increased in the distal part of colon (cloaca). Four types of endocrinocytes have been identified: EC, D, L and type IV. In frogs examined in March, endocrinocytes of the colon mucosal epithelium demonstrate structural features characteristic of the state of functional strain. These features are most pronounced in EC-cells. This seems to be due to the state of awakening from hibernation, serotonin (produced by EC cells) playing an essential role in this process. PMID- 14521085 TI - [Morphological changes in the kidney after 5/6 nephrectomy and low-dose radiation]. AB - In the present study, we investigated the effect of low-dose irradiation of experimental nephrectomized rats. We hypothized that the low-dose irradiation may slow down the development of focal-segmented glomerulosclerosis (FSGS) after 5/6 nephrectomy. Experiments were performed with 32 male Wistar rats, divided into four groups. The first group contained only operated animals. Animals in the second and third groups were irradiated on the next day after operation with 1 and 3 Gy, respectively. The healthy animals made the forth, control group. Attention was focused on physiological and morphological changes after low-dose (1 and 3 Gy) irradiation. We measured blood pressure, proteinuria, serum creatinin and cysC. Morphological changes of glomerulus and tubules were studied. Animals of the first group had significantly thicker glomerular basement membrane, compared to animals of other groups. The morphological study demonstrated degeneration of the tubular epithelium, tubular atrophy and FSGS. Besides, it was shown that changes in the third group (3 Gy) were less than in nephrectomized (first group) and 1 Gy (second group). The animals of the third group (3 Gy) had significantly lower proteinuria and FSGS. We conclude that our hypothesis, suggesting that low-dose irradiation slows down the development of FSGS, was confirmed. PMID- 14521087 TI - [Effect of alpha-tocopherol and synthetic antioxidants on morpho-functional status of gonadotropic cells from the adenohypophysis of albino rats]. AB - Morphometric indices of gonadotropic cells obtained from adenohypophysis of white rats, both males and females, were investigated after treatment with alpha tocopherol and synthetic antioxidants. The former stimulated the functional status of gonadotropic cells revealed in a proportional increase in both nuclear and cytoplasmic volumes. After the treatment with a synthetic antioxidant dibunolum, the volume of the cytoplasm increased in gonadotropic cells of rats of different sexes. After the treatment with a water-soluble antioxidant emoxipinum, the volume of the cytoplasm in gonadotropic cells increased only in males. The outcomes allow to consider alpha-tocopherol, in contrast to from synthetic antioxidants, as one of the modulators of the functional state of gonadotropic cells obtained from adenohypophysis. PMID- 14521086 TI - [The ultrastructure of apple cells during storage]. AB - A study was made of ultrastructural features of epidermal and hypodermal cells of freshly collected apples of early- and late-ripe cultivars, and of those storage at low temperature. In late-cultivars, the ultrastructure of most cells remained unchanged within 8 months (except increased vacuolation). In early-ripe cultivars, signs of senile destruction were evident in many hypodermal cells as early as within 2 months of storage. Apple cells of all cultivars contained a fairly developed smooth endoplasmic reticulum (SER). A suggestion is made that SER may be involved in the synthesis of abscisic acid essential for the fruit dormancy maintenance. PMID- 14521088 TI - [The role of phosphatidylinositol-3-kinases P85/P110 and hVPS34 in endocytosis of EGF-receptor complexes]. AB - The previous data (Zheleznova et al., 2001) did not enable the authors to conclude which particular wortmannin sensitive PI-3-kinase--p85/p110 (I class PI 3-K) or hVPS34 (III class PI-3-K)--may be involved in the regulation of EGF receptor endocytosis. In the present work, we have shown that upon stimulation of EGF-receptor endocytosis additional structures stained with antibody against p85 appear in A431 cells, but the p85-positive compartment never co-localized with EGF-receptor-containing compartments either in control or in wortmannin-treated cells. At the same time, wortmannin treatment prevented association of hVPS34 with endosomal membranes. We have also found that early endosomal markers--Rab5 and EEA1 (membrane association of the latter depends on Rab5 and hVPS34)--co localized with EGF-receptor in the juxtranuclear region during late stages of endocytosis, both in control and upon wortmannin treatment. These observations favor our suggestions that the transition of EGF-receptors from early to late endosomes may occur directly in this juxtranuclear region and be tightly associated with the formation of so called multivesicular bodies (MVB), which are late endosomes per se. We suggest that wortmannin may have no effect on early EEA1-dependent stage of the receptor endocytosis but blocks a transition of EGF receptor complexes into the late endosomes by inhibiting activity of hVPS34 and removing it from membranes. The hVPS34 product PI-3-K, according to the known data, is involved in the formation of internal vesicles of MVB. Accumulation of EGF-receptors in these vesicles is believed to be necessary for the receptor degradation. PMID- 14521089 TI - [Molecular-cytogenetic characteristics of B-chromosomes in chironomid (Diptera, Chironomidae)]. AB - Morphological and molecular study of B-chromosomes of three Chironomus species (siblings Ch. borokensis and Ch. phumosus from plumosus group, and Ch. heterodentatus from obtusidens group) was carried out. Morphological similarity of B-chromosome banding pattern and telomer-centromeric region banding pattern of chromosome IV in Ch. borokensis was shown. Polytene B-chromosomes of Ch. borokensis and Ch. heterodentatus were microdissected, and their DNA was amplified using degenerate oligonucleotide primer polymerase chain reaction. Comparative analysis of the localization of homologous B-chromosome DNA sequences of A- and B-polytene chromosomes was made using in situ fluorescence hybridization. It has been shown that B-chromosomes in the studied species are composed mainly of repetitive DNA sequences homologous to sequences of centromeric and telomeric DNA of A-chromosomes, and also these of the mobile element NLRCthl. The B-chromosome DNA, homologous to sequences of DNA mobile element, was scattered on A-chromosomes (more than 100 sites). No ribosomal DNA repeats were identified in B-chromosome. Heterologous FISH of B-chromosome DNA to polytene A-chromosomes of Ch. thummi, a species lacking B-chromosomes, enabled us to reveal the presence of numerous sites homologous to DNA of B-chromosomes. These are mainly mobile element sites. An origin of B-chromosomes and peculiarities of their organization in chironomids are discussed. PMID- 14521090 TI - [The role of protein kinase C in Na+ transport regulation in the skin of adult frogs and tadpoles of Rana temporaria]. AB - Using the voltage-clamp technique, a possible role of protein kinase C in regulation of Na+ transport in the skin of the frog Rana temporaria was investigated. It was shown that protein kinase C activator phorbol ester 12 myristate 13-acetate (PMA), applied to the apical surface of the skin, stimulated transepithelial Na+ transport, measured as amiloride-sensitive short-circuit current, and also increased such electrical characteristics of frog skin as the open-circuit potential and transepithelial conductance. PMA exerts a similar stimulation effect on Na+ transport across the tadpole skin. Specific inhibitors of protein kinase C, chelerythryne or H-7, almost fully prevented the PMA-induced stimulation of Na+ transport. These data support a concept that the response to PMA was indeed mediated by PKC activation. The results are compatible with the important role played by protein kinase C in regulation of transepithelial Na+ transport in the skin of R. temporaria. PMID- 14521091 TI - [Enhancement of growth promoting activity of human blood on keratinocytes after its irradiation in vivo (transcutaneously) and in vitro with visible and infrared polarized light]. AB - To stimulate wound healing, current medicine uses various methods of phototherapy. The induced activation of proliferative processes in the wound occurs due to development of not only local, but also systemic processes, whose nature remains largely uninvestigated. The present work provides evidences that as early as 30 min after irradiation of a small area of the volunteer's body surface with polychromatic visible light + infrared polarized light (400-3400 nm, 95% of polarization) at a therapeutic dose (12 J/cm2), soluble factors appear in the circulating blood, which are able to stimulate proliferation of human keratinocytes in primary culture. A similar effect was also revealed after a direct blood irradiation. A proof is provided in favor of a hypothesis that a rapid rise of growth promoting activity of the entire circulating blood may be a consequence of transcutaneous photomodification of the small amount of light modified blood in superficial skin vessels, and of the effect of such blood on its entire circulating volume. A possibility of a release into plasma of growth factors from blood cells and complexes with alpha 2-macroglobulin is discussed. PMID- 14521092 TI - [Metaphase chromosomes of human multiple myeloma cells of line U-266: karyotype and morpho-functional characteristics of nucleolar organizer regions]. AB - Human multiple myeloma (MM) cell lines are widely used to investigate chromosome rearrangements typical for this disease. However, during cell cultivation both numeral and structural chromosome rearrangements usually take place in addition to changes of structural and functional status of particular chromosome regions. We investigated karyotype and morpho-functional status of nucleolar organizer regions (NORs) in human cell line MM U-266. Cytogenetic analysis (G-banding) showed karyotypic stability and balanced chromosome set in hypodiploid (n = 44) U 266 cells. We found the presence of rearranged chromosomes, typical for U-266, which retained throughout many year cell cultivation. At the same time, further chromosome rearrangements were shown, along with a tendency of cells to polyploidization. Using FISH (rDNA-probe) and AgNOR-staining techniques, we found that only 4 of 8 NORS were Ag positive (AgNOR), whose dimensions varied from 1 to 3 units of arbitrary scale (u.a.s.). The average summarized AgNOR size was 7.19 +/- 0.03 u.a.s. Peculiarities of the NOR morpho-functional status in U-266 cells are discussed. PMID- 14521093 TI - You'll lose sleep over this pill. PMID- 14521095 TI - Making pro formas perform. AB - Regulators are trying to clear up the muddle created by earnings-report adjustments called "pro formas" that companies issue. Constraining such reporting, as the regulators seem bent on doing, isn't the solution. Firms should increase alternative reporting--and fully account for their accounting. PMID- 14521094 TI - Detroit's health-care crisis--and ours. PMID- 14521096 TI - And now, a word from our sponsor. AB - Bryant Pharmaceutical's flagship product, a popular arthritis medicine called Seflex, is selling well--but not well enough. With generic versions due on the shelves in a couple of years, the drug company is looking for a dramatic sales increase. No wonder marketing VP Laura Goldenberg feels the pressure. She knows she has to reach more consumers, but in an environment where people bombarded with advertising are using devices such as TiVo to skip ads, her job has suddenly taken on a new intensity. In search of a new, gangbuster campaign, Laura and her ad agency come up with the idea of product placement--not your typical integration of a product into a television or movie script, but a less traditional approach. Their idea is to hire a much-loved, elderly actress to extol the virtues of Seflex on a morning news program. The news segment would be about arthritis, and Seflex would be casually mentioned during the interview. The company would have to pay the actress $1 million, and there are risks: What if it gets out that Bryant is paying her? What if the actress errs and says something about Seflex's side effects? If something goes wrong, Bryant Pharmaceutical's reputation could suffer. Should the company green-light Laura's plan? This fictional case study looks at the pros and cons of traditional product placement and newer, more subtle alternatives to advertising. Commenting on the case are Bob Gamgort, president of Masterfoods USA; Michelle R. Nelson, an assistant professor of journalism and mass communications at the University of Wisconsin, Madison; FTC commissioner Mozelle W. Thompson; and Mike Sheehan, president and CEO of Hill, Holliday, Connors, Cosmopulos. PMID- 14521097 TI - Gilded and gelded. Hard-won lessons from the PR wars. AB - A golden statue of a winged youth once perched on the roof of AT&T's old headquarters. But when AT&T lowered the 24-foot-high statue for regilding so that it could be placed in the company's new headquarters, the chairman was shocked to discover that the figure was anatomically correct. So he decreed that it also be gelded. The altered "Golden Boy" thus became a metaphor for AT&T's recent embattled history, and it serves as a cautionary symbol for all companies operating in today's brutal business environment, where perception can be as important as reality. While image consultants and executives work to gild a company's image, special interest groups and the media can geld a company with countless little cuts. The author, a former executive vice president of public relations for AT&T, provides an insider's view of some of the company's most painful public-relations scrapes. They include the collapse of two apparent CEO succession plans, AT&T's inability to meet heightened expectations after Mike Armstrong was appointed CEO, and the racially charged furor over a cartoon in an employee publication. The author offers four lessons: Don't become hypnotized by your own buzz; understand the way the business media think; address the needs of all your stakeholders; and be sensitive to the possible emotional resonance of what appear to be straightforward facts. To illustrate the final point, the author mentions AT&T's elimination of 40,000 jobs in 1996. Wall Street was impressed, but NBC's Tom Brokaw said the workforce reduction might signal "another long, anxious year for the American middle class." No rational argument from AT&T could overcome the layoffs' symbolic impact. Wounded but wiser after numerous public-relations battles, the company eventually learned to stop aggregating job-reduction information for the media. PMID- 14521098 TI - The harder they fall. AB - The past decade may well be remembered as the era of the high-flying, aggressive leader. Corner-office titans like Kenneth Lay, Dennis Kozlowski, and Bernard Ebbers graced the covers of business magazines. They captured the public's fascination with their bold business moves and charismatic sound bites. Then scandal set in, and the stars fell to earth. In this article, social psychologist Roderick M. Kramer asks an important question: Why do so many leaders--not just in business, but also in politics, religion, and the media--display remarkable adeptness and ability while courting power, only to engage in even more remarkable bouts of folly once that power has been secured? Kramer, who has spent most of his career researching how leaders get to the top, says there is something about the process of becoming a leader that changes people in profound ways. The systems through which we select our leaders force executives to sacrifice the attitudes and behaviors that are essential to their survival once they have reached the top. Society has learned to consider risk taking and rule breaking as markers of good leadership. As a result, CEOs and other leaders lack the modesty and prudence needed to cope with the rewards and trappings of power. They come to believe that normal limits don't apply to them and that they are entitled to any spoils they can seize. The leaders who do remain grounded--who get to the top and stay there--exhibit five common psychological and behavioral habits: They simplify their lives, remaining humble and "awfully ordinary." They shine a light on their weaknesses instead of trying to cover them up. They float trial balloons to uncover the truth and prepare for the unexpected. They sweat the small stuff. And they reflect more, not less. PMID- 14521099 TI - The great transition. AB - As China's economy grows and opens further, the opportunity it presents to multinationals is changing. Foreign companies are moving to country development and new strategic choices. Now, foreign firms can actually go after the Chinese domestic market, and it's worth going after. Improvements in China's infrastructure, workforce, and regulatory environment are making it possible for companies to lower their costs to reap new competitive advantages. Multifaceted and often-shifting risks accompany this shifting opportunity. The reforms required for admission into the WTO will be politically difficult for China to implement, and its progress will be slowed by the scarcity of resources for the country's shaky banking system, the inadequacy of the social safety net, environmental problems, and local governments' cash shortage. China's breathtaking 9% average annual GDP growth rests on an unsteady foundation of overcapitalized state-owned enterprises, which have oversupplied many markets, and fiercely protectionist regional government officials pursuing growth-at almost-all-costs policies. Frequent changes in regulations, bureaucracies, and reporting relationships will continue to make planning difficult, and, as the SARS epidemic demonstrated, there is always the potential for serious disruptions. But for at least the next ten years, multinationals should be the biggest winners in China. To reap the benefits, a multinational must properly nest its effort into its overall organization, show "one face to China" at the national level but also tailor local strategies, be wary of joint ventures, and mitigate risk, in particular the theft of intellectual property. China is a major opportunity for companies that forthrightly face its complexities. It will remain largely inscrutable--and unprofitable--for the rest. PMID- 14521100 TI - The Chinese negotiation. AB - Most Westerners preparing for a business trip to China like to arm themselves with a list of etiquette how-tos. "Carry a boatload of business cards," tipsters say. "Bring your own interpreter." "Speak in short sentences." "Wear a conservative suit." Such advice can help get companies in the door and even through the first series of business transactions. But it won't sustain the prolonged, year-in, year-out associations Chinese and Western businesses can now achieve. The authors' work with dozens of companies and thousands of American and Chinese executives over the past 20 years has demonstrated that a superficial adherence to etiquette rules gets executives only so far. They have witnessed communication breakdowns between American and Chinese businesspeople time and time again. The root cause: the American side's failure to understand the much broader context of Chinese culture and values, a problem that too often leaves Western negotiators flummoxed and flailing. American and Chinese approaches often appear incompatible. Americans see Chinese negotiators as inefficient, indirect, and even dishonest, while the Chinese see American negotiators as aggressive, impersonal, and excitable. Such perceptions have deep cultural origins. Yet those who know how to navigate these differences can develop thriving, mutually profitable, and satisfying business relationships. Four cultural threads have bound the Chinese people together for some 5,000 years, and these show through in Chinese business negotiations. They are agrarianism, morality, the Chinese pictographic language, and wariness of strangers. Most Western businesspeople often find those elements mysterious and confusing. But ignore them at any time during the negotiation process, and the deal can easily fall apart. PMID- 14521101 TI - The hidden dragons. AB - Most multinational corporations are fascinated with China. Carried away by the number of potential customers and the relatively cheap labor, firms seeking a presence in China have traditionally focused on selling products, setting up manufacturing facilities, or both. But they've ignored an important development: the emergence of Chinese firms as powerful rivals--in China and also in the global market. In this article, Ming Zeng and Peter Williamson describe how Chinese companies like Haier, Legend, and Pearl River Piano have quietly managed to grab market share from older, bigger, and financially stronger rivals in Asia, Europe, and the United States. Global managers tend to offer the usual explanations for why Chinese companies don't pose a threat: They aren't big enough or profitable enough to compete overseas, the managers say, and these primarily state-owned companies are ill-financed and ill-equipped for global competition. As the government's policies about the private ownership of companies changed from forbidding the practice to encouraging it, a new breed of Chinese companies evolved. The authors outline the four types of hybrid Chinese companies that are simultaneously tackling the global market. China's national champions are using their advantages as domestic leaders to build global brands. The dedicated exporters are entering foreign markets on the strength of their economies of scale. The competitive networks have taken on world markets by bringing together small, specialized companies that operate in close proximity. And the technology upstarts are using innovations developed by China's government owned research institutes to enter emerging sectors such as biotechnology. Zeng and Williamson identify these budding multinationals, analyze their strategies, and evaluate their weaknesses. PMID- 14521102 TI - The real new economy. AB - During the soar-and-swoon days of the late 1990s, many people believed that information technology, and the Internet in particular, were "changing everything" in business. A fundamental change did happen in the 1990s, but it was less about technology than about competition. Under director Diana Farrell, the McKinsey Global Institute has conducted an extensive study of productivity and its connection to corporate IT spending and use during that period. The study revealed that information technology is important--but not central--to the fate of industries and individual companies. So if information technology was not the primary factor in the productivity surge, what was? The study points to competition and innovation. In those industries that saw increases in competitive intensity, managers were forced to innovate aggressively to protect their revenues and profits. Those innovations--in products, business practices, and technology--led to the gains in productivity. In fact, a critical dynamic of the new economy--the real new economy--is the virtuous cycle of competition, innovation, and productivity growth. Managers can innovate in many ways, but during the 1990s, information technology was a particularly powerful tool, for three reasons: First, IT enabled the development of attractive new products and efficient new business processes. Second, it facilitated the rapid industrywide diffusion of innovations. And third, it exhibited strong scale economies--its benefits multiplied rapidly as its use expanded. This article reveals surprising data on how various industries in the United States and Europe were affected by competition, innovation, and information technology in the 1990s and offers insights about how managers can get more from their IT investments. PMID- 14521103 TI - The lean service machine. AB - Jefferson Pilot Financial, a life insurance and annuities firm, like many U.S. service companies at the end of the 1990s was looking for new ways to grow. Its top managers recognized that JPF needed to differentiate itself in the eyes of its customers, the independent life-insurance advisers who sell and service policies. To establish itself as these advisers' preferred partner, it set out to reduce the turnaround time on policy applications, simplify the submission process, and reduce errors. JPF's managers looked to the "lean production" practices that U.S. manufacturers adopted in response to competition from Japanese companies. Lean production is built around the concept of continuous flow processing--a departure from traditional production systems, in which large batches are processed at each step. JPF appointed a "lean team" to reengineer its New Business unit's operations, beginning with the creation of a "model cell"--a fully functioning microcosm of JPF's entire process. This approach allowed managers to experiment and smooth out the kinks while working toward an optimal design. The team applied lean-manufacturing practices, including placing linked processes near one another, balancing employees' workloads, posting performance results, and measuring performance and productivity from the customer's perspective. Customer-focused metrics helped erode the employees' "My work is all that matters" mind-set. The results were so impressive that JPF is rolling out similar systems across many of its operations. To convince employees of the value of lean production, the lean team introduced a simulation in which teams compete to build the best paper airplane based on invented customer specifications. This game drives home lean production's basic principles, establishing a foundation for deep and far-reaching changes in the production system. PMID- 14521104 TI - Pharmacologic considerations in geriatric patients. PMID- 14521105 TI - HRT for elder with hot flashes? PMID- 14521106 TI - Testing for drugs of abuse. PMID- 14521107 TI - Evidence-based treatment of hypertension. JNC 7 Guidelines provide an updated framework. PMID- 14521108 TI - Parkinson's disease. More than meets the eye. PMID- 14521109 TI - Wrinkles and beyond. Skin problems in older adults. PMID- 14521110 TI - Terminal dementia in the elderly. Awareness leads to more appropriate care. PMID- 14521111 TI - Charitable giving. Challenging, frustrating and invigorating, volunteering is time well spent. PMID- 14521112 TI - Patient information. Preventing falls among older adults. PMID- 14521113 TI - Insurance coverage for reversible contraception. A legal battle for women and NPs. PMID- 14521114 TI - Arthropod bites and stings. Recognition and treatment. PMID- 14521115 TI - Caring for each other. PMID- 14521116 TI - A small fortune? The review body has agreed to the Agenda for Change pay rise- but there are issues yet to be resolved. PMID- 14521117 TI - HRT: the risks. What the recent Million Women Study means for patients. PMID- 14521118 TI - Change in practice. PMID- 14521119 TI - One pill fits all. PMID- 14521120 TI - MMR down, measles up. PMID- 14521121 TI - Production line. PMID- 14521122 TI - Fat-free targets. PMID- 14521123 TI - Breathing space. PMID- 14521124 TI - Reflective practice and team teaching in mental health care. AB - AIM: To develop and test new approaches to professional education that would address issues of team capacity and readiness to learn, thereby strengthening the ability to practise reflectively. METHOD: The time out and the debrief approaches to team training in core competencies were piloted with ten care teams in the north west of England. Pre- and post-training measures of competency were obtained to assess the effectiveness of the training. RESULTS: The results show that competence measures taken after attending the programme were significantly higher than those taken before training. Self-assessments and manager assessments supported this, indicating that the training had a positive effect on the overall capability of practitioners. CONCLUSION: The findings suggest that the innovative educational process created in this study could be used to support the structures of reflexivity and thoughtful practice. This educational process could be used to support practitioners to fully embrace work challenges. PMID- 14521125 TI - Disease-modifying drugs in multiple sclerosis. AB - This article is a description of practice and a comment on the evolution of a disease-modifying drug (DMD) treatment service for multiple sclerosis (MS) in a tertiary referral centre. It is hoped it will help other MS nurses who are involved in the establishment of DMD clinics in their area. It aims to offer an update on practical management of DMDs that may prove useful for any nurse involved in caring for people with MS. It describes lessons learnt in the establishment and growth of the service and protocols put in place to ensure best practice. The importance of partnership in care with patients and multidisciplinary collaboration is highlighted, while the pivotal role of the MS nurse in the delivery of an effective service is emphasised. PMID- 14521126 TI - Understanding the origins of practice problems. AB - Set against a background of modern-day nursing practice becoming ever more complex, this article aims to help nurses solve problems by understanding their origins' and recognising them when they occur. PMID- 14521127 TI - Back to the bed side. PMID- 14521128 TI - Taking it to the top. PMID- 14521129 TI - Should I become a vegetarian? PMID- 14521130 TI - Bring on the chocolates, Valentine! PMID- 14521131 TI - Understanding, preventing, and treating hypertension in women. PMID- 14521133 TI - By the way, doctor. I've had a pain in my shoulder for two weeks, which my doctor says is tendonitis of my rotator cuff. What can you tell me about this condition? I can't recall doing anything to cause the pain. PMID- 14521134 TI - [Nursing outcomes and the reform of care processes (III). A problem of maximal and minimal levels]. AB - This article presents a reflection about the different focus which exists between analysis of the role nurses play, investigating what nurses role is and about the product of nurses' professional activities, and the definition and evaluation of the nursing product. Nursing products approach viability, optimization and maximization of resources. From the tension between both aspects, the best possible nursing professional should develop. This article also presents an analysis of some of the systems used to evaluate the hospital product; among these are APACHE II, PMC, DS, CPHA, GRD, SIGNO. PMID- 14521132 TI - By the way, doctor. My gynecologist says that I have atrophic vaginitis and that it's a common problem for menopausal women. In all my reading about menopause, I've never come across this condition. What can you tell me about it? PMID- 14521135 TI - [3rd Regional Research Congress of Psychiatric and Neurologic Specialty Hospitals 24-26 October 2001]. PMID- 14521137 TI - Drug utilisation 90% profiles--a useful tool for quality assessment of prescribing in primary health care in Stockholm. AB - PURPOSE: To analyse strengths and weaknesses of a simple method for assessing the general quality of drug prescribing and to study the acceptance of the method among general practitioners (GPs). METHODS: Prescriptions dispensed during October-December 1999 and 2000, respectively, were analysed for 38 Primary Health Care centres (PHC) in Stockholm participating in an intervention project with the aim of increasing cost-consciousness among GPs. Focus was on quality of prescribing rather than on costs. Prescribing profiles focusing on the number of drugs constituting 90% of the volume (= DU90%) and the adherence to local drug committee guideline within this segment were presented for the prescribers. The credibility and usefulness of the method was evaluated by a questionnaire. RESULT: Among the PHCs, the total number of drugs prescribed varied between 358 and 674. The number of drugs in the DU90%-segment varied between 117 and 194. The adherence to guideline within this segment varied between 56% and 74% and increased over time. The prescribers found the DU90%-profiles clear and relevant and considered the method to be a useful tool for improving the quality of drug prescribing. CONCLUSION: Providing DU90%-profiles with guideline adherence as feedback was shown to be a valuable tool in general practice for assessing the overall quality in prescribing and to form the basis for more disease- or patient specific analyses. PMID- 14521136 TI - Fenton oxidation of cork cooking wastewater--overall kinetic analysis. AB - In the present work, the possibility of using chemical oxidation through Fenton's reagent for the pre-treatment of cork cooking wastewaters was exploited. Aiming both the selection of the best operating conditions (pH, Fe2+:H2O2 ratio and initial H2O2 concentration) and the evaluation of the overall reaction kinetics, trials were performed in a batch reactor. Operating at pH = 3.2, H2O2 concentration = 10.6 g/L and Fe2+:H2O2 ratio = 1:5 (by weight), about 66.4% of total organic carbon (TOC), 87.3% of chemical oxygen demand (COD) and 70.2% of biochemical oxygen demand (BOD5) were removed and an increase of the BOD5/COD ratio from 0.27 to 0.63 was achieved. In the temperature range 20-50 degrees C, the best performance was obtained at 30 degrees C. The kinetic study was undertaken at different initial TOC concentrations and temperatures. Overall kinetics can be described by a second-order followed by a zero-order rate equation and the apparent kinetic constants at 30 degrees C are k = 2.3 x 10(-4) L/mg min and k0 = 26.0 mg/L min, respectively. The experiments performed at different temperatures confirmed the global kinetic model and allowed to calculate the global activation energy for the second-order reaction (70.7 kJ/mol). PMID- 14521138 TI - [Quid? Cherubism]. PMID- 14521140 TI - Immigration and ancillary health care providers. PMID- 14521139 TI - [Comparison of piclamilast with ciclamilast in bronchodilating and antiallergic effects]. AB - OBJECTIVE: To compare the bronchofilating and antiallergic effects with piclamilast with ciclamilast, the second-generation phosphodiesterase 4 (PDE4) selective inhibitors. METHODS: Effects of piclamilast and ciclamilast on airway smooth muscle (ASM) at resting tension, carbachol-induced contraction and the synergistic effect of two agents on isoproterenol-induced bronchorelaxation were evaluated in the isolated tracheal strips of guinea pig in a cumulative manner in vitro. Slow reaction substance of anaphylaxis (SRS-A) release from lung tissues of the sensitized guinea pigs after antigen challenge was examined by bioassay. Antiallergic effect of piclamilast, ciclamilast and rolipram on the isolated ASM of sensitized guinea pigs were evaluated with Schultz-Dale reaction. RESULTS: Piclamilast and ciclamilast showed bronchorelaxant effect in ASM at resting tension. EC50 values of piclamilast and ciclamilast were 1.00 x 10(-5) mol/L and 0.84 x 10(-5) mol/L. Piclamilast and ciclamilast could both enhance the bronchodilating effect of isoproterenol in the isolated ASM of guinea pig, reduce the amount of SRS-A released from lung tissues of the sensitized guinea pigs and also inhibit ovalbumin (OA)-induced bronchoconstruction (Schultz-Dale reaction). CONCLUSION: The results indicate the bronchodilating effect of ciclamilast is as potent as piclamilast, but the antiallergic effect of ciclamilast is significantly more potent than that of piclamilast. PMID- 14521141 TI - Screening for developmental toxicity of tobacco smoke constituents. PMID- 14521142 TI - [Federal Recommendation Council adopts framework recommendation on quality assurance]. PMID- 14521144 TI - [Great attention should be paid to study of genetic susceptibility of liver cirrhosis]. PMID- 14521145 TI - [Seminar report. Section of Internal Medicine of the German Society of Internal Medicine May 23-26 in Bamberg]]. PMID- 14521146 TI - [Venous blood: echogenicity and ultrasound]. PMID- 14521147 TI - [An expanded OPS-3-1 catalog for sonography]. PMID- 14521148 TI - [ADR 2130: the first expanded commercial array instrument]. PMID- 14521149 TI - Simulation of abdomen sonography. Evaluation of a new ultrasound simulator. AB - AIM: We developed and evaluated a simulator for the sonography of the abdomen in order to improve the teaching quality in sonography training. METHOD: Eleven medicine residents who had received 4 to 12 months full time sonography training performed ultrasound examinations of the right upper quadrant in 5 consecutive patients and in 5 simulator cases. The correctness of their findings and the time required for the examinations were measured. The subjective confidence in their findings and the handling of the ultrasound machines were rated on a visual analogue scale. RESULTS: During patient ultrasound examination 75 % (SEM 9%) of all pathologic findings were recognized by the residents, whereas 71 % (SEM 8%) of the pathologies of the simulator cases were found. This minimal difference was not significant in the paired, two sided t-test (p = 0.15). Severe pathologies did not escape detection. The time required for patient examination(10.57 min, SEM 3.25 min) was not significantly different (p = 0.53) to the time required for the simulator cases (9.59 min, SEM 2.98 min). The subjective confidence in the sonographic findings did not differ significantly (p = 0.39) between the real patient situation (68%, SEM 6%) and the simulation (64%, SEM 12 %). Only the handling of the ultrasound machines was judged to be significantly better (p=0.008) than the simulator (74%,SEM 7% vs. 61 %, SEM 12%). CONCLUSION: In this first direct crossover comparison between real patient sonography and simulator based scanning we proved that the simulator we developed simulates the real patient examination reliably and reproducibly. PMID- 14521150 TI - Chronic recurring infarction of the spleen: sonographic patterns and complications. AB - PURPOSE: Splenic infarction is a major problem of splenic pathology but is characterized by a high tendency for complete healing. The purpose of this study is to describe frequency, sonographic patterns, and complications of chronic infarction (CI) METHODS: Between 1980-2001 550 patients with focal splenic lesions were diagnosed by ultrasound. Eighty patients had an acute infarction, and in 14 cases a chronic infarction was diagnosed and confirmed by cytohistology/splenectomy (n = 3) or sonographic follow-up examination (n = 11). All patients with Cl had been investigated by B-mode sonography and colour Doppler sonography (CDS). Data were retrospectively evaluated. RESULTS: Two types of Cl could be discriminated. Type I morphology (n =8) was predominantly found in homozygous sickle-cell anaemia (n= 6) and sonographically characterized by a small or normal sized spleen (n = 6), with diffuse enhanced echogenicity (n= 8), and foci with diminished echogenicity (n=5). Type II morphology (n = 6) was predominantly found in myeloproliferative diseases (n = 4) and characterized by an enlarged spleen with a homogeneous echotexture (n = 7), and a solitary (n = 6), triangular (n=4), hyperechoic (n=4) splenic foci near the splenic surface. On CDS CI were characterized by absent flow signals (n = 7) or by reduced flow signals (n= 7). Spontaneous splenic ruptures occurred as infarction related complications in 3 of 14 cases (21%). CONCLUSIONS: CI develops in 17.5% of patients with infarctions. It occurs predominantly in patients with sickle-cell anaemia and myeloproliferative disease. Two characteristic morphologic patterns were found and associated with an increased risk of spontaneous splenic rupture: Therefore sonographic follow-ups might be useful. PMID- 14521151 TI - Bilateral lesions in the basal ganglia of a patient with acquired immunodeficiency syndrome. PMID- 14521152 TI - Magnetic resonance imaging in foetuses with bilateral moderate ventriculomegaly and suspected anomaly of the corpus callosum on ultrasound scan. AB - OBJECTIVE: The diagnosis of anomalies of the corpus callosum (ACC) in foetuses with bilateral moderate ventriculomegaly (BMV) is difficult by means of ultrasound scan. The aim of this study was to examine the value of the additional investigation with magnetic resonance imaging (MRI) in foetuses with BMV and suspected ACC on ultrasound scan. Pathogenesis and clinical presentation of BMV and ACC are discussed. METHODS: 41 foetuses with central nervous system (CNS) anomalies on ultrasound scan were assessed by ultrasonography and MRI from 1999 to 2001. Eight of these 41 foetuses presented with BMV and suspected ACC on ultrasound scan and were prospectively included in the study. Foetal investigations with sonography and MRI were analysed with regard to diagnostic confidence; results were correlated with post partum findings. Six of these 41 foetuses presented with BMV without suspected ACC on ultrasound scan and were retrospectively analysed. RESULTS: Ultrasonography suspected ACC in 8 foetuses with BMV. MRI confirmed the presence of ACC in 4 of these 8 cases. MRI additionally showed ACC in two of the six retrospectively analysed foetuses with BMV without suspected ACC on ultrasound scan. Prenatal MRI diagnosis was confirmed after delivery in all cases. CONCLUSION: MRI is more sensitive than ultrasonography in the evaluation of ACC in foetuses with BMV. For prenatal screening ultrasound still remains the investigation of choice. PMID- 14521153 TI - [Mode-of-action of manual medicine in the cervical spine]. AB - Manual medicine aims at diagnosing and treating different disorders of the musculoskeletal system. It is a multidisciplinary approach with special emphasis on reversible disorders of the joints, muscles and ligaments. This treatment conception includes chirotherapy, physical therapy and drug treatment. The spine, an in particular the cervical spine, is treated primarily for joint disorders characterized by a variety of symptoms (e.g.headache, vertigo, dizziness, blurred vision). Manual medicine should be an integral part of modern clinical otolaryngology. PMID- 14521154 TI - Green tea epigallocatechin-3-gallate is an inhibitor of mammalian histidine decarboxylase. AB - (-)-epigallocatechin-3-gallate, an antiproliferative and antiangiogenic component of green tea, has been reported to inhibit dopa decarboxylase. In this report,we show that this compound also inhibits histidine decarboxylase, the enzymic activity responsible for histamine biosynthesis. This inhibition was proved by a double approach, activity measurements and UV-Vis spectra of enzyme-bound pyridoxal-5'-phosphate. At 0.1 mM (-)-epi-gallocatechin-3-gallate, histidine decarboxylase activity was inhibited by more than 60% and the typical spectrum of the internal aldimine form shifted to a stable major maximum at 345 nm, suggesting that the compound causes a stable change in the structure of the holoenzyme. Since histamine release is one of the primary events in many inflammatory responses, a new potential application of (-)-epigallocatechin-3 gallate in prevention or treatment of inflammatory processes is suggested by these data. PMID- 14521155 TI - Crystal structure of Enterobacter cloacae 908R class C beta-lactamase bound to iodo-acetamido-phenyl boronic acid, a transition-state analogue. AB - The structures of the class C beta-lactamase from Enterobacter cloacae 908R alone and in complex with a boronic acid transition-state analogue were determined by X ray crystallography at 2.1 and 2.3 A, respectively. The structure of the enzyme resembles those of other class C beta-lactamases. The structure of the complex with the transition-state analogue, iodo-acetamido-phenyl boronic acid, shows that the inhibitor is covalently bound to the active-site serine (Ser64). Binding of the inhibitor within the active site is compared with previously determined structures of complexes with other class C enzymes. The structure of the boronic acid adduct indicates ways to improve the affinity of this class of inhibitors. This structure of 908R class C beta-lactamase in complex with a transition-state analogue provides further insights into the mechanism of action of these hydrolases. PMID- 14521157 TI - Bibliography. Current world literature. Nose and paranasal sinuses. PMID- 14521156 TI - [Vascularized tumor of the scalp]. PMID- 14521158 TI - The healthy newborn partnership and the 'Dhaka Declaration'. PMID- 14521159 TI - The pre-senile delusion of infestation. PMID- 14521160 TI - Morselli's views on eugenics. PMID- 14521161 TI - Preventing childhood injuries. PMID- 14521163 TI - Bibliography. Current world literature. Head and neck oncology. PMID- 14521162 TI - Asthma. PMID- 14521164 TI - Bibliography. Current world literature. Head and neck reconstruction. PMID- 14521165 TI - Bibliography. Current world literature. Facial plastic surgery. PMID- 14521166 TI - Predialysis care in diabetic patients: the missing link? PMID- 14521167 TI - Peritoneal catheter exit-site infections: predisposing factors, prevention and treatment. AB - Catheter-related infections, exit-site-tunnel infections and peritonitis remain the Achilles heel of peritoneal dialysis. Although the overall incidence of peritoneal-dialysis-related infectious complications has been reduced since the introduction of the Y-set and double bag system, approximately one-fifth of peritonitis episodes are associated with catheter exit-site and tunnel infections. Since its development in 1968, the Tenckhoff catheter has become one of the most widely used peritoneal catheters, and many have proposed that a number of modifications have made it a better choice. Controversies concerning the effect on exit-site infections of catheter(s) with one or two cuffs, with straight, coiled, Swan-Neck, or other modifications led to the randomized controlled studies that are reviewed in this paper. Several studies have confirmed that mupirocin, applied at the exit-site as part of regular exit-site care, reduces the risk of S. aureus exit-site and tunnel infections. Recently, the emergence on a world-wide basis of mupirocin-resistant S. aureus (MuRSA) in peritoneal dialysis patients has brought this prophylactic strategy into question. However the low frequency of resistant organisms after four years of mupirocin prophylaxis suggests that we can continue its use with annual surveillance. Once established, exit-site infections may respond to appropriate treatment, but if not the only option may be catheter removal and replacement. Although peritonitis risk has decreased over the past decade, mainly due to improvements in connection technology, exit-site and tunnel infections have not. An exit-site infection that does not respond to treatment may lead to tunnel infection and to persistent peritonitis, which may require catheter removal and occasionally discontinuation of the peritoneal dialysis. Therefore it is important to be familiar with these factors that predispose to exit-site infection and to know how to prevent and to treat such infections. This review will discuss factors that predispose to catheter-related exit-site infections, techniques of exit-site care, and ways to prevent exit-site infection, with emphasis on S. aureus infections and their treatment. PMID- 14521168 TI - Clinical application of sodium profiling in the treatment of intradialytic hypotension. AB - BACKGROUND: Intradialytic hypotension is mainly induced by the removal of extracellular sodium during dialysis, which impairs intravascular fluid refilling and reduces blood volume. To counter this complication we tested a new kind of profiled hemodialysis (PHD) consisting of the intradialytic modulation of dialysate sodium concentration according to individual profiles set up using a new mathematical model for intradialytic solutes and water kinetics. The clinical aim of this PHD is to stabilize blood pressure maintaining higher blood volume values than standard dialysis treatments. We clinically validated PHD in comparison with constant dialysate sodium dialysis (CHD). METHODS: Twenty hypotensive dialysis patients underwent one PHD and one CHD session maintaining the same dialysis length, sodium mass removal and body weight decrease. A new mathematical model was used to define both the dialysate sodium profiles for PHD and the constant dialysate sodium for CHD. Percent blood volume variation (Crit line), mean blood pressure, heart rate, cardiac output (Doppler-echocardiography) were monitored intradialitically. RESULTS: Cardiovascular stability improved on PHD as compared with CHD sessions; blood volume and cardiac output during PHD showed a lower decrease than on CHD, the differences statistically significant (from 30' and 60' respectively). Mean blood pressure was, at all time intervals, more stable on PHD than on CHD and was accompanied, on PHD, by a lower heart rate increase (differences statistically significant). CONCLUSIONS: This study shows that PHD performed using dialysate sodium profiles elaborated by our mathematical model obtains, in hypotensive patients, a higher hemodynamic intradialytic stability than CHD, probably due to a higher stabilization of blood volume. PMID- 14521169 TI - Beta2-microglobulin clearance with super high flux hemodialysis: an ex vivo study. AB - BACKGROUND: Beta2m accumulation induces disease in patients with end-stage renal failure (ESRF). Thus, its removal from patients with ESRF appears desirable. Current dialysis technology, however, has limited effectiveness. AIMS: To measure beta2m clearance with a novel super high flux membrane. DESIGN: Ex vivo experimental study. SETTING: Intensive Care Laboratory of Tertiary institution. SUBJECTS: Six volunteers. MEASUREMENTS AND RESULTS: At a blood flow of 300 ml/min, the clearance of beta2-MG increased from 113.5 +/- 38.5 ml/min with a dialysate flow rate of 200 ml/min to 184.8 +/- 61.1 ml/min with a flow rate of 300 ml/min and 195.0 +/- 60.0 ml/min with a 500 ml/min flow rate. The clearance of albumin was 4.5 ml/min with a dialysate flow rate of 200 ml/min, 5.2 ml/min for a flow rate of 300 ml/min and 5.8 ml/min for a flow rate of 500 ml/min. CONCLUSIONS: High levels of beta2m clearance can be achieved with a super high flux membrane while albumin losses remain limited. PMID- 14521171 TI - A novel bioartificial liver containing small tissue fragments: efficiency in the treatment of acute hepatic failure induced by carbon tetrachloride in rats. AB - The efficiency of a new bioartificial liver (BAL) containing small tissue fragments in the treatment of acute hepatic failure induced by carbon tetrachloride in rats was evaluated. A day after injection (i.p.) of CCl4 the animals were connected to a BAL containing liver fragments (fragment BAL) and a BAL containing no liver fragments (no-fragment BAL), and extracorporeal hemoperfusion was carried out for 4 h. The activities of alanine transaminase and lactate dehydrogenase as well as the concentrations of ammonia, glucose, urea, and amino acids in plasma were measured. A tendency to the stabilisation of ammonia, glucose, phenylalanine, tyrosine, and other amino acids was revealed at the end of hemoperfusion in poisoned rats connected to the fragment BAL. A statistically significant difference in survival between the animals connected to the fragment BAL and no-fragment BAL was found. The results obtained indicate that the bioreactor containing small liver fragments is effective in the treatment of acute hepatic failure in animals. PMID- 14521170 TI - Computerized generation of circadian sensor modal days with continuous glucose monitoring for comparison of various insulin regimens based on insulin glargine in type 1 diabetes. AB - AIM: Our aims were (1) to design and standardize a statistical approach for data reduction in continuous glucose monitoring, allowing comparison of circadian glycemic patterns in therapeutic subcohorts of patients with type 1 diabetes, and (2) to investigate the applicability of this approach for CGMS assessment in clinical study of basal insulin replacement quality with various timings of basal injections (pre-breakfast, dinner, bedtime) of a new insulin analog. METHODS: Prospective randomized three-arm parallel study with switch over after 6 months for another 3 months of free choice injection time point (options pre-breakfast, pre-dinner and bedtime) of the new insulin analog in 16 type 1 diabetic subjects on functional insulin treatment (FIT: basal, prandial and correctional dosages). CGMS was used at the end of each follow up period of a clinical study. Representative daily profiles were off-line computed as "circadian sensor modal days" for each insulin regimen consisting of consecutive means of hourly glucose values. RESULTS: Although the overall quality of glycemic control (HbAIC) for different regimens did not reach statistical differences, CGMS displayed slightly divergent maximal swings in the course of glycemia (p=0.04-0.08) and allowed- with delineated data reduction procedure--a reliable between treatment comparison. CONCLUSION: Off-line computation of "hourly circadian sensor modal days" for data reduction can be effectively used with CGMS for description of circadian glycemic patterns in type 1 diabetes. PMID- 14521172 TI - In vitro evaluation of the PUCA II intra-arterial LVAD. AB - The "pulsatile catheter" (PUCA) pump is a minimally invasive intra-arterial left ventricular assist device intended for acute support of critically ill heart failure patients. To assess the hydrodynamic performance of the PUCA II, driven by an Arrow AutoCat IABP driver, we used a (static) mock circulatory system in which the PUCA II was tested at different loading conditions. The PUCA II was subsequently introduced in a (dynamic) cardiovascular simulator (CVS) to mimic actual in vivo operating conditions, with different heart rates and 2 levels of left ventricular (LV) contractility. Mock circulation data shows that PUCA II pump performance is sensitive to afterload, pump rate and preload. CVS data demonstrate that PUCA II provides effective LV unloading and augments diastolic aortic pressure. The contribution of PUCA II to total flow is inversely related to LV contractility and is higher at high heart rates. We conclude that, with the current IABP driver, the PUCA II is most effective in 1:1 mode in left ventricles with low contractility. PMID- 14521174 TI - Estimation of coupled assist flows in a moving-actuator bi-ventricular assist device using interventricular pressure. AB - An assist flow estimation scheme for a moving-actuator biventricular assist device (MA-BVAD) using interventricular pressure (IVP) has been developed. The scheme uses a waveform feature parameter of IVP, peak IVP time (PIT), for estimation of the filling volumes of both left and right blood sacs simultaneously. In a regression analysis on data from an in vivo test in an 85 Kg male calf for 20 days, the PIT was found to have high correlation with the blood sac filling volume (R=0.883: left filling volume, R=0.967: right filling volume). A conceptual equation hypothesizing this correlation between PIT and filling volume was established based on the observation and the unknown parameters were identified using least squares parameter optimization. The estimation equation identified proved highly accurate (R=0.916 for left flow, R=0.970 for right flow). The accuracy of the estimation scheme promises very good practical applicability. PMID- 14521173 TI - Hemofiltration during cardiopulmonary bypass for high risk adult cardiac surgery. AB - AIMS: The role of hemofiltration (HF) during cardiopulmonary bypass (CPB) in adult cardiac surgery is controversial. It may be beneficial during prolonged CPB in high-risk surgery. Accordingly, we sought to compare two groups of patients undergoing high-risk cardiac surgery with or without HF. METHODS: One hundred and eighteen patients who underwent complex cardiac surgical procedures during a 12 month period were divided into two groups. Group I (n=61) comprised patients who were treated with hemofiltration during CPB. Group II (n=57) were not filtered. Estimated risk of death, standard demographic, clinical and surgical features were obtained and predetermined outcomes were studied. Statistical comparisons were made. RESULTS: Age, procedure times and mortality rates were similar in both groups. The mean volume of fluid removed in group I was 3.4 L. The preoperative mean Parsonnet score was 24.8 in group I and 22.5 in group II (ns). Postoperative serum hemoglobin, hematocrit, platelet, and albumin levels were all significantly higher in group I patients (p=0.0015) indicating hemoconcentration. Post operative chest drainage showed a trend toward decreased post-operative bleeding in group I (p=0.065). Postoperative pleural effusions requiring chest tube drainage were significantly less in group I (9.8% vs. 29.8% 6; p = 0.0062). The incidence of lung infection was also decreased from 26.3% to 13.1% (p=0.05). Operative mortality was similar in both groups (11.4% in group 1, 10.5% in group II, ns). CONCLUSION: Hemofiltration during CPB attenuates postoperative anemia, thrombocytopenia and hypoalbuminemia, may reduce post-operative bleeding and appears to decrease post-operative pulmonary complications. PMID- 14521175 TI - A collagen-based scaffold for a tissue engineered human cornea: physical and physiological properties. AB - Stabilized collagen-glycosaminoglycan scaffolds for tissue engineered human corneas were characterized. Hydrated matrices were constructed by blending type I collagen with chondroitin sulphates (CS), with glutaraldehyde crosslinking. A corneal keratocyte cell line was added to the scaffolds with or without corneal epithelial and endothelial cells. Constructs were grown with or without ascorbic acid. Wound-healing was evaluated in chemical-treated constructs. Native, noncrosslinked gels were soft with limited longevity. Crosslinking strengthened the matrix yet permitted cell growth. CS addition increased transparency. Keratocytes grown within the matrix had higher frequencies of K+ channel expression than keratocytes grown on plastic. Ascorbic acid increased uncrosslinked matrix degradation in the presence of keratocytes, while it enhanced keratocyte growth and endogenous collagen synthesis in crosslinked matrices. Wounded constructs showed recovery from exposure to chemical irritants. In conclusion, this study demonstrates that our engineered, stabilized matrix is well-suited to function as an in vitro corneal stroma. PMID- 14521176 TI - In vitro biocompatibility of titanium oxide for prosthetic devices nanostructured by low pressure metal-organic chemical vapor deposition. AB - Metal-Organic Chemical Vapor Deposition (MOCVD) has recently been proposed to coat orthopedic and dental prostheses with metal nanostructured oxide films through the decomposition of oxygenated compounds (single-source precursors) or the reaction of oxygen-free metal compounds with oxygenating agents. The present study was carried out to assess the in vitro biocompatibility in terms of cell proliferation and activation, of commercially pure Ti (control material: TI/MA) coated with nanostructured TiO2 film by MOCVD (Ti/MOCVD) using osteoblast-like cell cultures (MG-63). Evaluations were performed at 3, 7 and 14 days. Cell proliferation showed a similar trend for Ti/MA and TilMOCVD compared to polystyrene; cell number increased with time from seeding to day 7 (p < 0.005), and then decreased progressively until day 14 (ranging from -14% to -47%). The ALP level and OC production showed no significant differences between Ti/MOCVD and Ti/MA at each experimental time. Significantly higher ALP levels were found in Ti/MA at 3 days and in Ti/MOCVD at 7 and 14 days when compared to the polystyrene group. OC production decreased over time and the highest values were observed at 3 days, when it was significantly higher in the Ti/MA than in the polystyrene group (50%, p < 0.05). CICP synthesis was positively affected by the presence of Ti/MOCVD and was higher in Ti/MOCVD than in the polystyrene group. No significant differences were found between Ti/MOCVD and Ti/MA in terms of IL-6 and TGF-beta1 synthesis at any experimental time. In conclusion, the current findings demonstrate that the nanostructured TiO2 coating positively affects the osteoblast-like cell behavior in terms of cell proliferation and activity, thus confirming its high level of in vitro biocompatibility in accordance with expectations. PMID- 14521177 TI - Personality disorder symptoms in adolescence: a five-factor model perspective. AB - The five-factor model has been widely used to describe adaptive and maladaptive functioning in adulthood. However, less is known about the structure and developmental antecedents of personality pathology and personality disorders. In the present study, we examined the validity of the most recent DSM-IV predictions (Widiger, Trull, Clarkin, Sanderson, & Costa, 2002) in a sample of 419 non clinical adolescents and explored the validity of the unique FFM facetvariances (using the NEO PI-R) to predict disorder symptoms (using the ADP-IV). Our results demonstrate a largely similar correlation and regression pattern between adult and adolescent data, indicating that adaptive and maladaptive trait-descriptive systems relate across a more extended developmental span than has been demonstrated before. PMID- 14521178 TI - Self-reported personality traits and disorders (DSM-IV) and risk of criminal recidivism: a prospective study. AB - Assessment and management of criminal offenders require valid methods to recognize personality psychopathology and other risk and protective factors for recidivism. We prospectively explored the association between dimensional and categorical measures of personality disorder (PD) measured with the DSM-IV and ICD-10 Personality Questionnaire (DIP-Q, Ottosson et al., 1995) and registered reconvictions in adult offenders. One hundred and sixty-eight offenders consecutively referred for pre-sentencing forensic psychiatric evaluation in Sweden during 1995-1996 completed DIP-Q self-reports. The subjects received different types of sanctions and were followed for an average of 36 months after release from prison, discharge from a forensic psychiatric hospital, or onset of nondetaining sentences. Age-adjusted odds ratios revealed a 4.8 times higher risk for any recidivism and a 3.7 times higher risk for violent recidivism among subjects whose self-reports suggested a categorical diagnosis of antisocial PD as compared to offenders without antisocial PD. The remaining nine categorical DSM IV PD diagnoses were not significantly related to recidivism. In dimensional analyses, each additional antisocial and schizoid PD symptom endorsed by participants at baseline increased the risk for violent reoffending. Our results suggest a relationship between self-reported behavioral instability and interpersonal dysfunction captured primarily by DSM-IV antisocial and schizoid PD constructs, and criminal re-offending also in a multi-problem sample of identified offenders. PMID- 14521179 TI - Psychopathology of incarcerated sex offenders. AB - The psychopathology and particularly the personality disorders of sex offenders were compared to general inmates of the Colorado Department of Corrections. Using the MCMI-III (Millon, 1994, 1997), sex offenders in general were found to have more varied types of personalities than general population inmates. Specifically, they were more schizoid, avoidant, depressive, dependent, self-defeating, and schizotypal. General population inmates had the more classically criminal personality characteristics of antisocial, narcissistic, and sadistic. Multivariate analysis showed the Dependent, Narcissistic, Antisocial, and Schizotypal scales to be the most differentiating. Sex offenders were also found to have more affective psychopathology such as anxiety, dysthymia, PTSD, and major depression. A similar trend was found when comparing child molesters to rapists. The child molesters were more neurotic, affective, and socially impaired than the rapists. Multivariate analysis showed the Dependent scale to be the most important in differentiating these two types of sex offenders. PMID- 14521180 TI - Cognitive vulnerability to depression in individuals with borderline personality disorder. AB - The goal of the current study was to examine whether individuals with comorbid Major Depressive Disorder (MDD) and Borderline Personality Disorder (BPD) exhibit greater severity of depressive symptoms than (1) individuals with MDD without BPD and (2) individuals with neither MDD nor BPD. One hundred and forty-one individuals participated in a semi-structured clinical interview assessing MDD and BPD. They also completed measures assessing depressive symptoms, depressogenic attributional style, hopelessness, self-esteem, rumination, and dysfunctional attitudes. In line with hypotheses, individuals with BPD and MDD exhibited higher levels of depressive symptoms and cognitive vulnerability than individuals in the other two groups. In addition, after controlling for the effects of cognitive vulnerability, the effect of group membership on depressive symptoms was reduced, suggesting that the increased severity of depressive symptoms experienced by those with BPD is partially due to their possessing higher levels of cognitive vulnerability to depression. PMID- 14521181 TI - Convergent validity of the MCMI-III personality disorder scales and the MMPI-2 scales. AB - The MCMI-III personality disorder scales (Millon, 1994) were empirically validated in a sample of prisoners, psychiatric inpatients, and outpatients (N = 477). The scale intercorrelations were congruent with those obtained by Millon, Davis, and Millon (1997). We conclude that our Flemish/Dutch version shows no significant differences with the original version of the MCMI-III as far as intercorrelations are concerned. Convergent validity of the MCMI-III personality disorder scales was evaluated by the correlational data between the MCMI-III personality disorder scales and the MMPI-2 clinical (Butcher, Dahlstrom, Graham, Tellegen, & Kaemmer, 1989) and personality disorder (Somwaru & Ben-Porath, 1995) scales. Improved convergence was obtained compared with previous versions of the MCMI-I. Only the compulsive MCMI-III personality disorder scale remains problematic. The scale even showed negative correlations with some of the related clinical scales and with the corresponding personality disorder scales of the MMPI-2. PMID- 14521182 TI - Cluster C personality disorder and recovery from major depression: 24-month prospective follow-up. AB - We investigated whether Cluster C personality disorder (CPD) is associated with recovery from depression. Changes in symptom scales in 30 patients with MD and CPD were compared with changes in 60 patients with MD alone over a 24-month follow-up period. Recovery of patients with MD and comorbid CPD was inferior to recovery of those with MD alone based on the Hamilton and Beck Depression Inventory (BDI) scales, and the SCL-90 total score. Only 18% of those with MD alone but 47% of those with CPD and MD met the criteria for major depression at the end of the 24-month follow-up. Multiple logistic regression analyses revealed an independent association between the lack of recovery (BDI score > 9 at 24 months) and the presence of CPD (OR 4.9, 95% CI 1.5-16.0). Moreover, the presence of CPD associated with the presence of major depression at 24 months (OR 4.2, 95% CI 1.4-12.2). The presence of CPD hinders the alleviation of depressive symptoms in major depression. PMID- 14521183 TI - Assessing personality disorders using a systematic clinical interview: evaluation of an alternative to structured interviews. AB - The aim of this study was to assess interrater reliability and provide initial data bearing on the validity of a method of assessing personality disorders (PDs) that does not presume that patients can accurately self-report personality pathology. In a sample of 24 outpatients, two clinician-judges independently applied the Shedler-Westen Assessment Procedure-200 (SWAP-200; Westen & Shedler, 1999a, 2000), a 200-item Q-sort procedure for assessing personality pathology, to data from the Clinical Diagnostic Interview (Westen, 2002), a systematic clinical interview that mirrors and standardizes methods used by experienced clinicians to diagnose personality. In 16 of the 24 cases, the treating clinician also independently described the patient using the SWAP-200 Q-sort, based on longitudinal knowledge of the patient over the course of treatment, blind to the interview data. Interrater reliability was uniformly high, with median correlations between interviewers at r > .80. Interviewer-treating clinician correlations were also high, with median convergent validity coefficients at r > .80. Diagnostic overlap (discriminant validity) was moderate for dimensional DSM IV diagnoses, reflecting extensive comorbidity among disorders, but minimal for empirically derived diagnoses identified in prior research. Treating clinicians' dimensional PD diagnoses using this method also strongly predicted interviewer rated measures of adaptive functioning. The findings provide preliminary support for the reliability and validity of an alternative to structured interviews for diagnosing personality pathology, and suggest that the way to improve validity of personality diagnosis may not be to minimize clinical inference but to quantify it using psychometric instruments. PMID- 14521184 TI - Introduction: autism--the challenges ahead. PMID- 14521185 TI - Epidemiology and early identification of autism: research challenges and opportunities. AB - Recent studies suggest that the prevalence of autism spectrum disorders may be as high as 60 per 10000, considerably greater than the long-accepted figure of 5 per 10 000 for classic autism. Increased recognition, the broadening of the diagnostic concept and methodological differences across studies may account for most or all of the apparent increase in prevalence, although this cannot be quantified. In addition to the implications for families and services, these conceptual changes will affect the scientific study of autism. At present, case definition is reliant on the behavioural and developmental picture alone. Because the behavioural phenotype of autism and the broader autism spectrum disorders includes individuals with different ultimate aetiologies, even when biological or genetic markers are found they will not be present in all individuals with the phenotype. The fact that autism is not a unitary 'disorder' presents a significant challenge to genetic, biological, neurological and psychological research. Progress has recently been made in the earlier identification of autism both through screening programmes and by increased understanding and enhanced surveillance. This offers an opportunity to better understand the early developmental course of autism and may provide additional clues to the underlying pathology. PMID- 14521186 TI - Implications of the broader phenotype for concepts of autism. AB - Autism, like many new diseases, was initially characterized by its most severe phenotypic manifestation and the ability to explain these distinctive features has been the benchmark against which explanatory models have subsequently been judged. Our understanding of the significance of milder phenotypes in other relatives has shifted from presumed environmental aetiological factors to variable manifestations of a complex disease process. In this paper we outline how the challenge of explaining the full range of phenotypic expression inevitably leads to more complex models of disease process than previously supposed. The implications of milder phenotypes for genetic, neurobiological and cognitive models of autism will be considered in relationship to several key features of complex diseases: complexity, hierarchy, emergence and coherence. PMID- 14521187 TI - Strategies for autism candidate gene analysis. AB - The identification of autism susceptibility genes has moved a step closer over the last four years with the completion of eight whole genome screens for linkage. Several overlapping areas of linkage have been reported, most notably on chromosomes 7q22-31 and 2q32. These regions of replicated linkage provide a focus to search for candidate genes whose normal functions in neurodevelopment are altered to increase the risk for autism. Strategies that aim to narrow further the rather broad size of these linkage regions, such as high density single nucleotide polymorphism (SNP)-based association studies, currently suffer from practical and statistical limitations. Alternatively, positional candidate genes can be screened for deleterious variants in autistic individuals selected from large samples such as those collected by the International Molecular Genetic Study of Autism Consortium (IMGSAC). Targeted genotyping of candidate gene variants in this large multiplex family sample will then be performed to confirm association with autism. PMID- 14521188 TI - How might genetic mechanisms operate in autism? AB - Twin and family studies provide strong evidence that autism has a largely genetic aetiology. The pattern of familial aggregation suggests that in individual families, a small number of genes act together to cause the phenotype. However, it is unlikely that the same genes act in all families. Thus, the total number of genes involved could be large. One key to finding genes for disorders with considerable locus heterogeneity is to detect genetically more homogeneous subsamples. There exist several traits in families who have a child with autism- biochemical, physical, or behavioural--that are likely to reflect underlying genetic heterogeneity and can thus be used to divide families into more homogeneous subsets. These traits (1) show variation in autism samples; (2) are found in non-autistic family members more often than controls; (3) aggregate in particular autism families; and (4) result in increased signals when used in linkage analysis to define 'affected'. PMID- 14521189 TI - X-linked genes and the neural basis of social cognition. AB - The neural basis of social cognition is subject to intensive research in both humans and non-human primates. Autism is the archetypal disorder of social cognitive skills, and increasing interest is being paid to the role played by efferent and afferent connectivity between the amygdala and neocortical brain regions, in predisposing to this condition. Such circuits are now known to be critical for the processing of social information. Recent research suggests a sub cortical neural pathway, routed through the amygdala, may turn out to be a key player in the mystery of why humans are so prone to disorders of social adjustment. This pathway responds to certain simple classes of potential threat, including direct eye contact and, in humans, arousal evoked by this exquisitely social stimulus is modulated and controlled by a variety of specific (largely frontal) neocortical regions. Dysfunction of these modulating circuits can occur in the context of developmental disorders that are associated with haploinsufficiency of one or more classes of X-linked genes, lacking Y homologues, which may be sexually dimorphic in expression. PMID- 14521190 TI - The neuropathology of the autism spectrum disorders: what have we learned? AB - Autism is a behaviourally defined disorder, initially described by Kanner in 1943. By definition, symptoms are manifested by 36 months of age and are characterized by delayed and disordered language, impaired social interaction, abnormal responses to sensory stimuli, events and objects, poor eye contact, an insistence on sameness, an unusual capacity for rote memory, repetitive and stereotypic behaviour and a normal physical appearance. Relatively few neuropathological studies have been performed on the brains of autistic subjects. Of those reported, abnormalities have been described in the cerebral cortex, the brainstem, the limbic system and the cerebellum. Although those with the disorder present with a specific set of core characteristics, each individual patient is somewhat different from another. Thus, it should not be surprising that the brains of these subjects should show a wide range of abnormalities. However, it is important to delineate the anatomic features, which are common to all cases, regardless of age, sex and IQ, in order to begin to understand the central neurobiological profile of this disorder. The results of our systematic studies indicate that the anatomic features that are consistently abnormal in all cases include reduced numbers of Purkinje cells in the cerebellum, and small tightly packed neurons in the entorhinal cortex and in the medially placed nuclei of the amygdala. It is known that the limbic system is important for learning and memory, and that the amygdala plays a role in emotion and behaviour. Research in the cerebellum indicates that this structure is important as a modulator of a variety of brain functions and impacts on language processing, anticipatory and motor planning, mental imagery and timed sequencing. Defining the differences and similarities in brain anatomy in autism and correlating these observations with detailed clinical descriptions of the patient may allow us greater insight into the underlying neurobiology of this disorder. PMID- 14521191 TI - Microbiology and immunology of autism spectrum disorders. AB - Both generic and environmental factors are likely to contribute to the pathogenesis of neurodevelopmental disorders. Even in heritable disorders of high penetrance, variability in timing of onset or severity of disease indicate a role for modifying principles. Investigation in animal models of the consequences of interactions between host response genes and microbes, toxins, and other environmental agents in a temporal context may elucidate the pathophysiology of a wide spectrum of chronic diseases. Here we review the evidence that infectious and immune factors may contribute to the pathogenesis of neurodevelopmental disorders, describe an animal model of neurodevelopmental disorders based upon viral infection, identify processes by which neural circuitry may be compromised, and outline plans for translational research in animal models and prospective human birth cohorts. PMID- 14521193 TI - The amygdala, autism and anxiety. AB - Brothers has proposed that the amygdala is an important component of the neural network that underlies social cognition. And Bauman and Kemper observed signs of neuropathology in the amygdala of the post-mortem autistic brain. These findings, in addition to recent functional neuroimaging data, have led Baron-Cohen and colleagues to propose that dysfunction of the amygdala may be responsible, in part, for the impairment of social functioning that is a hallmark feature of autism. Recent data from studies in our laboratory on the effects of amygdala lesions in the macaque monkey are at variance with a fundamental role for the amygdala in social behaviour. If the amygdala is not essential for normal social behaviour, as seems to be the case in both non-human primates and selected patients with bilateral amygdala damage, then it is unlikely to be the substrate for the abnormal social behaviour of autism. However, damage to the amygdala does have an effect on a monkey's response to normally fear-inducing stimuli, such as snakes, and removes a natural reluctance to engage novel conspecifics in social interactions. These findings lead to the conclusion that an important role for the amygdala is in the detection of threats and mobilizing an appropriate behavioural response, part of which is fear. If the amygdala is pathological in subjects with autism, it may contribute to their abnormal fears and increased anxiety rather than their abnormal social behaviour. PMID- 14521192 TI - What do imaging studies tell us about the neural basis of autism? AB - There is no clear evidence from imaging studies for specific structural abnormalities in the brains of people with autism. The most robust observation is of greater total brain volume. There is evidence that this greater volume is not present at birth, but appears during the first few years. This brain enlargement might be a marker of abnormal connectivity due to lack of pruning. While abnormalities have often been reported in the cerebellum and the amygdala, these are difficult to interpret since both increases and decreases in the size of these structures have been observed. Another way of identifying the neural basis of autism is to investigate brain systems underlying cognitive functions compromised in this disorder such as face perception and 'theory of mind'. Autistic people fail to activate the 'fusiform face area' during face perception tasks and show weak activation of medial frontal cortex and superior temporal gyrus when performing theory of mind tasks. These problems stem from a lack of integration of sensory processing with cognitive evaluation. I speculate that this problem reflects a failure of top-down modulation of early sensory processing. The problem could result from abnormal connectivity and lack of pruning. PMID- 14521194 TI - Cognition in autism: one deficit or many? AB - The aim of this paper is to provoke discussion concerning the nature of the cognitive impairments that characterize autism. Autism spectrum disorders appear to be heterogeneous at the biological and behavioural levels, but it is currently unclear whether one or more cognitive abnormalities may be universal to people with autism. In addition, it is unknown whether one cognitive deficit is primary and causal, or whether several complimentary accounts are needed to explain the full range of behavioural features. From research to date, it seems that the psychological abnormalities that characterize autism may be dissociable, and it is uncertain whether the degree of social and non-social impairments is related. Possible reasons for the co-occurrence of social and non-social cognitive abnormalities in autism are discussed. One implication is that searching for the biological bases of specific social and non-social deficits may be more profitable than searching for the aetiology of autism per se. PMID- 14521196 TI - Why have drug treatments been so disappointing? AB - The title of this contribution involves two consecutive questions: have the effects of medication in autism indeed been disappointing? And if so, why? The answer to the first question depends on whether one focuses on the core social and communicative deficits of autism, or on various complicating behaviour problems. Attempts over the past decades to develop drugs that specifically improve social and communicative functioning have failed. Among the most ambitious attempts were medical interventions in the endogenous opioid system that were motivated from animal models on the involvement of this system in various aspects of social behaviour. By contrast, medications such as the newer antipsychotics, psychostimulants, presynaptic noradrenergic blocking agents (clonidine and guanfacine) and selective serotonin reuptake inhibitors were shown to reduce impairing complicating symptoms of affective instability, irritability, hyperactivity and inattentiveness, aggression, self-injury and stereotypies. The explanation for the medication-refractory status of social and communicative deficits should be sought in at least two related factors: (1) the as yet unidentified neurochemical basis of autism, and (2) the obvious lack of involvement of the main neurotransmitter systems (dopamine, noradrenaline and serotonin) in the pathophysiology of social and communicative behaviour. PMID- 14521195 TI - Autism and specific language impairment: categorical distinction or continuum? AB - Traditionally, autism and specific language impairment (SLI) are regarded as distinct disorders, with differential diagnosis hinging on two features. First, in SLI one sees isolated language impairments in the context of otherwise normal development, whereas in autism a triad of impairments is seen, affecting communication, social interaction and behavioural repertoire. Second, there are different communication problems in these two conditions. Children with SLI have particular difficulty with structural aspects of language (phonology and syntax). In contrast, abnormal use of language (pragmatics) is the most striking feature of autism. However, recently, this conventional view has been challenged on three counts. First, children with autism have structural language impairments similar to those in SLI. Second, some children have symptoms intermediate between autism and SLT. Third, there is a high rate of language impairments in relatives of people with autism, suggesting aetiological continuities between SLI and autism. One interpretation of these findings is to regard autism as 'SLI plus', i.e. to assume that the only factor differentiating the disorders is the presence of additional impairments in autism. It is suggested that a more plausible interpretation is to regard structural and pragmatic language impairments as correlated but separable consequences of common underlying risk factors. PMID- 14521197 TI - Can early interventions alter the course of autism? AB - Interventions for autism have come a long way since the condition was described by Kanner in the 1940s. At that time, autism was considered to be closely linked to schizophrenia, and inadequate parenting was viewed as the principal cause. Psychoanalysis was often the therapy of choice, but there was also widespread use of the drugs and even electroconvulsive treatments that had been developed for use in schizophrenia. Over the years, as autism has come to be recognized as a developmental disorder, interventions have focused instead on enhancing developmental skills and on ways of ameliorating behavioural difficulties. Recognition of the role that language deficits in particular play in causing behaviour problems has led to a focus on the teaching of more effective communication skills. The need for early support for families and appropriate education is also widely acknowledged. Nevertheless, follow-up studies indicate that the prognosis for the majority of individuals with autism remains poor. And despite claims to the contrary, there is little evidence that very early, intensive interventions can significantly alter the long-term course of the disorder. The paper discusses findings from follow-up studies over the years and assess the impact of different intervention procedures on outcome. PMID- 14521198 TI - Early intervention and brain plasticity in autism. AB - Autism is associated with impairments in brain systems that come on line very early in life. One such system supports the development of face processing. Dawson and colleagues found that 3 year old children with autism failed to show differential event-related potentials (ERPs) to photographs of their mother's versus a stranger's face. Since differential ERP activity to familiar and unfamiliar faces is typically present by 6 months, this represents early brain dysfunction. McPartland and colleagues found that the face-specific ERP component ('N170') is atypical in older individuals with autism. N170 is typically larger to faces than non-faces, and prominent over the right hemisphere. In individuals with autism, N170 was larger for furniture than faces and bilaterally distributed. Biology and experience contribute to the development of face processing systems. Newborns are capable of recognizing faces. Early face recognition abilities are thought to be served by a subcortical system, which is replaced by an experience-dependent cortical system. Development of a neural system specialized for faces may depend on experience with faces during an early sensitive period. Because children with autism fail to attend to faces, they might not acquire the expertise needed for a specialized face processing system to develop normally. Early interventions that enhance social attention should result in changes in brain activity, as reflected in ERPs to face stimuli, with those children showing the greatest social improvement exhibiting more normal brain activity. PMID- 14521199 TI - Technical considerations in renal CT. AB - CT is a robust, rapid means of evaluation for a wide spectrum of urologic disorders. The evaluation of renal trauma, urologic malignancy, urolithiasis, and vascular anatomy is well suited to CT techniques. Subtle adjustments in the technical parameters and timing of the study, however, can optimize the evaluation based on the clinical setting. As CT is more widely used, often repeatedly on an individual patient, radiation exposure must be minimized while still obtaining diagnostic image quality. PMID- 14521200 TI - MR techniques for renal imaging. AB - This article describes the principles, attributes, and pitfalls of the many MR imaging approaches available for assessment of renal-related disorders. Tables 1 and 2 summarize the specific approach and rationale. PMID- 14521201 TI - Evaluation of renal causes of hypertension. AB - There are many renal causes of hypertension. Although RAS is the most common, other renal lesions can result in hypertension. Any evaluation of the kidney for hypertension should take all of these potential renal etiologies into consideration. PMID- 14521202 TI - Evaluation of the hypertensive infant: a rational approach to diagnosis. AB - This article reviews the literature and describes a methodologic approach to the diagnosis of hypertension in the young infant. The numerous etiologies of hypertension have been discussed and normative blood pressure data for neonates and infants have been provided. Techniques for accurate blood pressure measurement in the intensive care setting and for routine outpatient settings, are discussed. The lengthy discussion of radiologic approach to imaging can be summarized with the following suggested algorithm. Initial screening should be performed with gray-scale sonography, to identify renal parenchymal or collecting system abnormalities, including mass lesions and congenital anomalies. Further imaging with color and duplex Doppler sonography detects renal arterial or aortic thrombosis, and alterations in the arterial waveform caused by intrinsic or extrinsic renal artery narrowing. The major limitation of Doppler sonography is the recognition that disease in accessory renal arteries or in small segmental intrarenal arteries may frequently be undetected. Functional imaging with ACEI renography should follow renal sonography to detect hemodynamically significant renovascular disease (with a sensitivity and specificity of approximately 90%); intravenous enalaprilat is the preferred ACEI. Angiography should be reserved for older children in whom interventional percutaneous angioplasty may be more feasible. A young infant with hypertension caused by renal artery stenosis should be controlled medically until he or she is large enough to undergo angiography and angioplasty successfully. CT angiography and MR angiography, although promising in the adult population, may not adequately resolve the small intrarenal vessels, which are frequently the culprit in renovascular hypertension of infancy. PMID- 14521203 TI - CT urography and MR urography. AB - CT urography and MR urography are an evolving concept and developing technique. As the technology matures, CT urography will combine the ultimate diagnostic capabilities of intravenous urography and CT. In the near future, many intravenous urograms will be replaced by CT urography to evaluate patients with hematuria and other genitourinary conditions. MR urography currently serves as an alternative imaging technique to intravenous urography and CT urography for children and pregnant women and for patients with contraindications to iodinated contrast media. PMID- 14521204 TI - Renal imaging with ultrasound contrast: current status. AB - The application of UCAs to the kidney is still in its infancy; however, there are several areas of great promise. UCAs may replace CT in complex renal cyst evaluation and follow-up, eliminating the need for costly CT scans with their attendant potential contrast nephrotoxicity. This approach may decrease patient and physician uncertainty and improve diagnostic confidence. The use of UCAs is likely to be clinically useful in the evaluation of the indeterminate small renal mass on CT or MR imaging. Another probable useful application will be in renal artery stenosis. Routine application of UCAs may increase the percentage of diagnostic examinations, increase diagnostic confidence, and decrease examination times. It also will likely become the first line of evaluation in pyelonephritis, and be useful in immediate assessment of residual tumor after radiofrequency ablation. Of course, substantial additional work needs to be performed in large groups of patients to prove this currently optimistic outlook. PMID- 14521205 TI - CT evaluation of urinary lithiasis. AB - Unenhanced CT has been demonstrated to be the most accurate and efficient diagnostic imaging means to evaluate urinary lithiasis, with capability of directing management, and has become well accepted by radiologists, urologists, and emergency department physicians such that it is now the standard of practice. It is the duty of the radiologist to be aware of proper technique and the details of interpretation. The radiologist also has a duty to be aware of the limitations of unenhanced CT for detection and evaluation of various nonstone disorders, particularly with poor patient selection, and to extend the examination if appropriate. Controversies and future developments include cost containment with care for the selection of patients. Further attempts to reduce radiation exposure should be made. Optimal CT technique is not needed in general merely to detect urinary lithiasis. A consensus should be developed regarding use of CT in pregnant patients. Further improvements in the digital scout view would be useful for following patients. PMID- 14521207 TI - Imaging of renal trauma. AB - Trauma is a major cause of death and disability and renal injuries occur in up to 10% of patients with significant blunt abdominal trauma. Patients with penetrating trauma and hematuria, blunt trauma with shock and hematuria, or gross hematuria warrant imaging of the urinary tract specifically and CT is the preferred modality. If there is significant perinephric fluid, especially medially, or deep laceration, delayed images should be obtained to evaluate for urinary extravasation. Most renal injuries are minor, including contusions, subcapsular and perinephric hematoma, and superficial lacerations. More significant injuries include deep lacerations, shattered kidney, active hemorrhage, infarctions, and vascular pedicle and UPJ injuries. These injuries are more likely to need surgery or have delayed complications but may still often be managed conservatively. The presence of urinary extravasation and large devitalized areas of renal parenchyma, especially with associated injuries of intraperitoneal organs, is particularly prone to complication and usually requires surgery. Active hemorrhage should be recognized because it often indicates a need for urgent surgery or embolization to prevent exsanguination. PMID- 14521206 TI - MR imaging of renal function. AB - MR imaging is the only single noninvasive test that can potentially provide a complete picture of renal status with minimal risk to the patient, simultaneously improving diagnosis while lowering medical costs by virtue of its being a single test. The strengths of MR imaging lie in its high spatial and temporal resolution and its lack of exposure to ionizing radiation and nephrotoxic contrast agents. This article reviews the use of MR imaging for quantification of renal functional parameters and its application to clinical problems, such as RVD, hydronephrosis, and renal transplantation. Although advances in both the technical and clinical aspects of functional renal MR imaging have been made, much remains to be done. The preliminary results reported in the many studies reviewed are exciting, but these techniques need to be validated against accepted standards where such standards exist. In addition, and perhaps more important, the effects of these new diagnostic methods on patient outcomes must be studied. Finally, further progress in image processing and analysis must be made to make functional renal MR imaging truly practical. With these advances, one can expect functional renal MR imaging to play an ever-expanding and influential role in the care and management of the patient with renal disease. PMID- 14521208 TI - Imaging of hereditary renal cancer. AB - Over the past 5 years there have been dramatic developments in the extent of knowledge of hereditary renal cancers. In addition to VHL, which is associated with clear cell carcinoma, one can now list HPRC (associated with type I papillary renal cancer) and HLRCC (associated with type II papillary renal cancer). BHD and FRO are associated with chromophobe carcinoma and oncocytomas, although other histologic tumor types have been found in BHD. Medullary carcinoma of the kidney is associated with sickle cell trait. Although the genes associated with these tumors have been discovered, the exact mechanisms by which they cause renal cancer remain to be elucidated. It is quite likely that other genes also are involved in this process. Using VHL as an example, research is now underway on targeting mutant pVHL or excess HIF for diagnostic and therapeutic purposes. Understanding the mechanisms leading to cancer may open new targets of opportunity for drug development. This improved knowledge of the biogenetic pathways used to form tumors will impact the development of new therapeutic techniques for treating renal cancers in hereditary and nonhereditary forms of the disease. PMID- 14521209 TI - Surgical management of renal tumors. AB - Renal cell carcinoma is a relatively rare tumor, accounting for approximately 3% of malignancies in adults, but is the most common tumor of the kidney and the third most common tumor seen by urologists. Renal cell carcinoma is refractory to most traditional oncologic treatments, including chemotherapy, radiation therapy, and hormonal therapy. Because of recent advances in sophisticated radiologic studies, the surgeon can now make an accurate preoperative assessment of the nature and extent of kidney tumors. When evaluating renal tumors, the urologist looks for certain information to help in constructing a management plan. This article explores some of the points that contribute in the surgical decision making. PMID- 14521210 TI - Percutaneous image-guided radiofrequency ablation of renal malignancies. AB - There is a growing body of experience supporting the use of image-guided RFA for the treatment of primary RCC. Because surgical resection is a technique with low mortality, and a proved success rate that is high, this must remain standard therapy for patients with potentially curable RCC. Some patients with low-stage RCC, however, may not be surgical candidates. Image-guided RFA is an option for treatment of these patients. In addition, image-guided RFA shows promise for the successful care of other patients with RCC. In particular, RFA has been used successfully for the treatment of intractable hematuria resulting from an RCC; local recurrences of RCC, both for attempted cure and for palliation of symptoms; and finally for the treatment of isolated metastases from RCC. As with the treatment of primary RCC, the data remain limited for these applications. This technique should be reserved until after standard therapies have been exhausted. It seems likely that some form of image-guided percutaneous tumor therapy, such as RFA, will become an alternative treatment modality in some patients with potentially curable RCC. PMID- 14521211 TI - Factorial structure of the kidcope in Hong Kong adolescents. AB - The Kidcope (A. Spirito, L. J. Stark, & C. Williams, 1988), a brief screening measure of coping strategies for children and adolescents, was subjected to exploratory and confirmatory factor analyses in a sample of adolescents in Hong Kong. A 2-factor model was found to fit the data when only the 2 emotional regulation items were treated separately and were allowed to load on the 2 different factors. Further analyses showed that factor loadings, factor variances, and factor covariance were invariant across age and gender. On the basis of the factor analyses, the authors created 2 composite scores, representing control-oriented and escape-oriented coping strategies. Implications for the use of the Kidcope in research and preventive intervention are discussed. PMID- 14521212 TI - The relationship of the family social environment, peer influences, and peer relationships to altruistic orientation in Chinese children. AB - This study investigated the correlation between the family social environment, peer influences, and peer relationships to altruistic orientation in Chinese children. Results in the present study showed that (a) altruistic orientation measured by the Child Altruism Inventory (H. K. Ma & M. C. Leung, 1991) was directly associated with a positive family social environment, (b) altruistic orientation was directly associated with positive peer influences and was inversely associated with negative peer influences, and (c) altruistic orientation was directly associated with perceived prosocial behavior of one's best friend and inversely associated with perceived antisocial behavior of one's best friend. The findings suggested that a good family social environment, positive peer influences, and good peer relationships tended to increase altruistic orientation. PMID- 14521213 TI - Warmth and contingency and their relationship to maternal attitudes toward parenting. AB - In this study, the authors addressed the question of whether maternal parenting behaviors in terms of sensitivity, warmth, and contingency with respect to small infants were equally represented in attitudes toward parenting. Sixty mothers from central Germany with middle-class socioeconomic backgrounds were videotaped with their 1st born 3-month-old infant in a free-play and a caregiving episode in their home. The different maternal parenting components were behaviorally analyzed. Maternal attitudes toward parenting were assessed with self-report measures. The results showed that behavioral sensitivity was significantly associated with attitudes related to contingency, whereas behavioral warmth and contingency were not associated with maternal attitudes. The authors interpreted differential associations as a reflection of intuitive and explicit parenting strategies, with only sensitivity being explicitly represented. PMID- 14521214 TI - The development of the conception of socioeconomic mobility in children from Mexico and Spain. AB - The authors studied the developmental stages of children's understanding of upward socioeconomic mobility. They interviewed one hundred 6- to 14-year-old participants from Mexico and Spain and asked them about sources of wealth and factors related to socioeconomic mobility. Categorical analyses of the responses showed few age-related changes but noted some cross-national differences. A different analysis designed to identify levels of understanding showed a significant association between age and type of explanation of socioeconomic mobility. Overall, cross-national comparisons yielded similarities in children's developmental trends, and only slight differences were found with regard to cultural background. The present results contrast with those of studies conducted from the perspective of the social representation theory. PMID- 14521215 TI - The stroop color-word interference test as an indicator of ADHD in poor readers. AB - The performance on the Stroop Color-Word Interference Test of 36 boys with Attention Deficit Hyperactivity Disorder (ADHD) was compared with performances of a matched control sample. The control group outperformed their counterparts on the control and interference conditions of the Stroop test, suggesting ADHD specific executive and reading deficits. When individuals with both ADHD and reading disorders were excluded from the analysis, the authors found a significant difference between the ADHD group and the control group on the color word test, indicating that poor reading skills may produce false negatives on the Stroop test. However, fast and slow readers with ADHD did not perform differently from each other on the color-word test. The authors postulated the existence of two different causes of reading problems: phonological deficits and attentional deficits. PMID- 14521216 TI - A stage is a stage is a stage: a direct comparison of two scoring systems. AB - L. Kohlberg (1969) argued that his moral stages captured a developmental sequence specific to the moral domain. To explore that contention, the author compared stage assignments obtained with the Standard Issue Scoring System (A. Colby & L. Kohlberg, 1987a, 1987b) and those obtained with a generalized content-independent stage-scoring system called the Hierarchical Complexity Scoring System (T. L. Dawson, 2002a), on 637 moral judgment interviews (participants' ages ranged from 5 to 86 years). The correlation between stage scores produced with the 2 systems was .88. Although standard issue scoring and hierarchical complexity scoring often awarded different scores up to Kohlberg's Moral Stage 2/3, from his Moral Stage 3 onward, scores awarded with the two systems predominantly agreed. The author explores the implications for developmental research. PMID- 14521217 TI - UVB-induced mutations in human key gatekeeper genes governing signalling pathways and consequences for skin tumourigenesis. AB - The UVB component of the solar spectrum induces DNA lesions that, in the absence of error-free DNA repair, may give rise during DNA replication to mutations in caretaker and gatekeeper genes. The DNA repair genes are the best candidates for caretaker genes as exemplified by the human hereditary xeroderma pigmentosum (XP) syndrome. Cultured XP cells are hypermutable after UVB irradiation. This increased mutation frequency is also found in gatekeeper genes, which govern signalling pathways implicated in the control of cellular proliferation, differentiation and survival of human epidermal keratinocytes. We describe and discuss the role of mutated gatekeeper genes in five specific signalling pathways which have been implicated in skin carcinogenesis. The pathways we focus on in this review are: (i) P16(INK4A)-CDK4/6-RB; (ii) P14(ARF)-HDM2-P53; (iii) Sonic hedgehog (SHH)/GLI; (iv) WNT/beta-catenin; and (v) Bone Morphogenetic Protein (BMP)/SMAD. 70-80% of XP skin cancers exhibit one or several mutations in the P53, PTCH-1, SMO or CDKN2A genes, the type and frequency of mutated genes being different between squamous cell (SCCs) and basal cell carcinomas (BCCs). In XP cancers, the typically UVB-induced CC to TT tandem transitions represent approximately 60% of total mutations compared to 10-15% in skin tumours from DNA repair-proficient patients. Acquired activation of the pathways described herein can alter proliferation and differentiation of keratinocytes, allowing a damaged cell to replicate and give rise to mutated daughter cells, then eventually to the development of the carcinogenic process following clonal selection. PMID- 14521218 TI - Photochemical reactivity of polyenes: from dienes to rhodopsin, from microseconds to femtoseconds. AB - In reviewing the photochemistry of polyenes (from dienes and trienes to the visual retinyl chromophore), we categorize condensed-phase photochemical reactivity of molecules into two groups: those from thermally equilibrated excited species and those from unequilibrated excited species. Classical theories on radiationless transitions are useful for rationalizing the reactivity of molecules belonging to the first set only. The second group includes many of the exciting ultrafast photochemical reactions reported recently for polyenes (including dienes and trienes), in some cases with rates faster than vibrational relaxation. Much of the excited singlet-state reactions of polyenes, including the Hula-twist mechanism of photoisomerization, have been integrated with concepts introduced in other ultrafast spectroscopic/photochemical studies. Taking into consideration the special environment of the retinyl chromophore in rhodopsin, we propose a new mechanism for the phototrigger that accounts for its unusually fast rate of isomerization. PMID- 14521219 TI - Synthesis and isomerization of azobenzene dendrimers. AB - 3,3',5,5'-Tetramethoxyazobenzene and related meta-substituted azobenzene dendrimers show the first evidence of the dendritic effect on trans-to-cis photoisomerization; the thermodynamic parameters for cis-to-trans isomerization have been determined. PMID- 14521220 TI - Photosensitiser-controlled regioselectivity in the electron-transfer cycloreversion of 2,3-diphenyloxetanes. AB - The regioselectivity of the oxidative electron-transfer cycloreversion of trans,trans-2,3-diphenyl-4-methyloxetane is photosensitiser dependent. With chloranil the reaction proceeds through the trans-beta-methylstyrene radical cation; however, in the case of pyrylium salts, the trans-stilbene radical cation is formed. PMID- 14521221 TI - Modulation of macrophage structure and function by low level He-Ne laser irradiation. AB - Studies have shown that He-Ne laser irradiation can affect the biological functions of macrophages. The aim of the present study was to investigate the effect of He-Ne laser irradiation on the various functional parameters of macrophages and look for possible correlations in the effects to understand the mechanisms involved. Mice peritoneal macrophages were irradiated with a He-Ne laser (632.8 nm, approximately 10 W m(-2)) at energy densities ranging from 100 to 600 J m(-2) and the activities of lysozyme and cathepsin, phagocytosis, and cell spreading (markers of cell activation), as well as changes in NAD(P)H autofluorescence, were monitored. He-Ne laser irradiation was observed to lead to significant changes in all the parameters investigated. While lysozyme activity and spreading of the peripheral membrane were found to increase with the irradiation dose over the dose range investigated, the phagocytotic activity of macrophages, the activity of cathepsin, the observed decease in cell membrane fluidity and the observed increase in NAD(P)H level showed a peak at 200 J m(-2). Possible reasons for and the significance of the observed correlations are discussed. PMID- 14521222 TI - Enhanced UV-mediated free radical generation; DNA and mitochondrial damage caused by retinol supplementation. AB - Retinoid supplementation has been therapeutically used against various human disorders. We and others have demonstrated that retinol treatment causes free radical generation and increased iron uptake, iron storage and oxidative damage, both in vitro and in vivo. Here, we investigate the possible synergistic effect of retinol on UV-mediated free radical generation, oxidative damage to biomolecules and decreased cellular viability in primary cultured mammalian cells. Retinol treatment (7 microM) resulted in a threefold increase in UV mediated free radical generation and a 40%, increase in lipoperoxidation. DNA fragmentation and mitochondrial oxidative damage also increased significantly in retinol-supplemented UV-irradiated cultured cells as compared to UV-irradiated control cells, which were only treated with the solvent used to deliver the retinol (0.1% ethanol). All measurements were restored to control values when an iron chelator, 1,10-phenanthroline (100 microM), or an OH* scavenger, mannitol (1 mM), was co-administrated. Rather than protecting against free radical generation, retinol seems to enhance UV-mediated oxidative damage and decreases cellular viability in cultured cells. We suggest that retinol-enhanced iron uptake and storage and increased reactive oxygen species generated by the Fenton reaction may act synergistically with UV-irradiation in causing oxidative damage to cells. PMID- 14521224 TI - Assessing sublethal effects of ultraviolet radiation in juvenile rainbow trout (Oncorhynchus mykiss). AB - This paper presents a method for assessing sublethal effects of ultraviolet radiation in fish. The method is based on the assumption that UV-B exposure is stressful to fish. We measured oxygen consumption in UV-B-irradiated rainbow trout juveniles using a UV-transparent respirometer chamber. In addition, we simultaneously monitored fish behavior by videotaping. We found a direct relationship between UV-B irradiance and the percentage increase in oxygen consumption. Increased swimming activity and restless behavior were also noted under UV-B irradiation. The actual mechanisms by which fish perceive UV radiation and a behavioral reaction is triggered are unknown. Based on the present study, a response to UV-A irradiation cannot be ruled out. PMID- 14521223 TI - Optical properties and contribution of cuticle to UV protection in plants: experiments with apple fruit. AB - To assess the UV-screening capacity of plant surface structures, the optical properties of isolated cuticle and detached peel of apple fruit (Malus domestica Borkh., cv. Antonovka) have been studied. It was found that the cuticle exhibits considerable scattering of UV radiation, negligible absorption between 500-800 nm and attenuates UV radiation: on average, cuticular transmittance of non-reflected light amounts to 35.7 +/- 20.2 and 14.2 +/- 7.1% at 375 and 300 nm, respectively. The principal UV-A absorbers in the cuticle were identified as quercetin glycosides with an in vivo absorption maximum near 375 nm and content ranging from 10 to 70 nmol cm(-2). On the shaded side of apple fruit, both UV-A and UV-B absorption by the peel is, to a large extent, governed by cuticular phenolics, whereas on the sunlit surface, the absorption of the peel in the UV-A range is determined mainly by vacuolar peel flavonoids. It is concluded that a massive build-up of flavonoids in the peel cells located just below the cuticle, resulting in trapping of radiation in a broad spectral range, plays a dominant role in the long-term adaptation of apple fruit to elevated levels of solar radiation. PMID- 14521226 TI - Photooxygenation of allylic alcohols: kinetic comparison of unfunctionalized alkenes with prenol-type allylic alcohols, ethers and acetates. AB - The kinetics of the chemical and physical quenching of the first excited singlet state of oxygen [1O2 (1delta(g))] by unfunctionalized alkenes 1-4, allylic alcohols 5-7 and 9, allylic acetates 8 and 11, and the allylic ether 10 display small solvent-polarity effects on the reactivity. The regioselectivity of the singlet oxygen ene reaction is solvent independent for the unfunctionalized alkenes as well as the prenol-type substrates, the latter showing substantial solvent effects on the diastereoselectivity. Pronounced physical quenching is detected only for the allylic alcohols 5 and 6. These results are interpreted in terms of the interactions between singlet oxygen and the allylic hydroxy groups, conformationally promoted by allylic strain which lead either to chemical activation or to physical quenching. The results for substrate 9 in deuterated v.s non-deuterated methanol are in accord with hydrogen bonding between the allylic alcohol and 1O2, which directs the diastereoselectivity of the ene reaction with chiral allylic alcohols. PMID- 14521225 TI - EPR studies of the photodynamic properties of a novel potential photodynamic therapeutic agent: photogeneration of semiquinone radical anion and active oxygen species (O2*-, OH*, H2O2 and 1O2). AB - Cyclohexylamino-substituted hypocrellin B (CHAHB) has been synthesized with the aim of improving the red absorption and specific affinity for malignant tumors over those of the parent compound. Irradiation of a deoxygenated DMSO solution of CHAHB generates a strong electron paramagnetic resonance (EPR) signal, which is assigned to the semiquinone radical anion of CHAHB with the aid of a series of experimental results. In the presence of oxygen, superoxide radical anions (O2*-) are generated via electron transfer from CHAHB*-, the precursor, to ground-state molecular oxygen. Hydroxyl radicals were detected by spin-trapping EPR when an oxygen-saturated aqueous solution containing CHAHB and DMPO was irradiated. Singlet oxygen (1O2) is produced via energy transfer from triplet CHAHB to ground state oxygen molecules, with a sharply decreased quantum yield, i.e. 0.11. Furthermore, cell survival studies reveal CHAHB exhibits much higher photodynamic activities than its parent hypocrellins. The strongly enhanced photodynamic activities and sharply decreased quantum yield of 1O2 generation suggest that the type I (free radical) mechanism may play a significant role in CHAHB-PDT, rather than the type II (singlet oxygen) mechanism found in photofrin-PDT. PMID- 14521227 TI - Kinetic study of the oxidation of phenolic derivatives of alpha,alpha,alpha trifluorotoluene by singlet molecular oxygen [O2(1delta(g)) and hydrogen phosphate radicals. AB - The oxidation kinetics and mechanism of the phenolic derivatives of alpha,alpha,alpha-trifluorotoluene, 2-trifluoromethylphenol, 3 trifluoromethylphenol (3-TFMP), 4-trifluoromethylphenol and 3,5 bis(trifluoromethyl)phenol, mediated by singlet molecular oxygen, O2(1delta(g)), and hydrogen phosphate radicals were studied, employing time-resolved O2(1delta(g)) phosphorescence detection, polarographic determination of dissolved oxygen and flash photolysis. All the substrates are highly photo-oxidizable through a O2(1delta(g))-mediated mechanism. The phenols show overall quenching constants for O2(1delta(g)) of the order of 10(6) M(-1) s(-1) in D2O, while the values for the phenoxide ions in water range from 1.2 x 10(8) to 3.6 x 10(8) M( 1) s(-1). The effects of the pH and polarity of the medium on the kinetics of the photo-oxidative process suggest a charge-transfer mechanism. 2-Trifluoromethyl 1,4-benzoquinone is suspected to be the main photo-oxidation product for the substrate 3-TFMP. The absolute rate constants for the reactions of HPO4*- with the substrates range from 4 x 10(8) to 1 x 10(9) M(-1) s(-1). The 3 trifluoromethylphenoxyl radical was observed as the organic intermediate formed after reaction of 3-TFMP with HPO4*-, yielding 2,2' bis(fluorohydroxymethyl)biphenyl-4,4'-diol as the end product. The observed results indicate that singlet molecular oxygen and hydrogen phosphate radicals not only react at different rates with the phenols of alpha,alpha,alpha trifluorotoluene, but the reactions also proceed through different reaction channels. PMID- 14521228 TI - Continuous irradiation and flash-photolysis studies of new[3H]naphtho[2,1 b]pyrans linked by covalent bonds to oligothiophene units. Effect of thiophene substituents on the photochromism. AB - The influence of oligothiophene substituents on five new [3H]naphtho[2,1-b]pyrans has been investigated using flash photolysis and continuous irradiation techniques. Photochromic properties strongly depend on the number and position of the thienyl units. Thienyl substitution increases the UV/visible absorbance of the lower energy transition of both the naphthopyran and merocyanine isomers. The rate constants of the thermal fading processes also increase with the number of thiophenic entities. Although several minor photoisomers, whose relative amounts depend on the irradiation conditions could be involved, it has been assumed that under continuous monochromatic irradiation, the evolution of the photochromic reaction can be described by the formation of only one photoisomer. Under these conditions, quantum yields of photocolouration and molar absorption coefficients of the non-isolable photomerocyanine isomer were evaluated by photokinetic analysis. Flash-photolytic colourability can be interpreted using continuous irradiation parameters. A linear correlation has been established between experimental and calculated colourability. This result shows that similar photochromic reactions leading to similar photoisomers are triggered either by polychromatic flash photolysis or by continuous monochromatic irradiation. PMID- 14521229 TI - The quenching of indolic compounds by monosubstituted benzenes and the photoreaction with chlorobenzene. AB - The quenching of the excited singlet of indole and its methyl derivatives by monosubstituted benzenes has been studied in three solvents of different polarities and bimolecular quenching rate constants have been determined. Below the diffusion limit the rate constants decrease when solvent polarity increases, with the exception of those for the quenching by benzonitrile. For the latter the rate constants are near the diffusion limit in all cases. The decrease may be understood in terms of a lower energy of the indole excited state in the more polar solvents. In cyclohexane the remnant emission in the presence of high concentration of the quencher is clearly red shifted, indicating the presence of a new emitting species that can be ascribed to the presence of an exciplex. In some cases the emission of the exciplex can be clearly separated from the molecular fluorescence. In the quenching by chlorobenzene in ethyl acetate and acetonitrile a new highly fluorescent product is formed. This photoreaction is observed with all the indole derivatives and the kinetics of the reaction was followed by the increase in fluorescence intensity at a wavelength were the reactants do not emit. Three major products were identified as phenyl substituted indoles by GC-MS and their fluorescence emission and excitation spectra. PMID- 14521230 TI - Role of salicylic acid in systemic resistance induced by Pseudomonas fluorescens against Fusarium oxysporum f. sp. ciceri in chickpea. AB - Selected isolates of Pseudomonas fluorescens (Pf1-94, Pf4-92, Pf12-94, Pf151-94 and Pf179-94) and chemical resistance inducers (salicylic acid, acetylsalicylic acid, DL-norvaline, indole-3-carbinol and lichenan) were examined for growth promotion and induced systemic resistance against Fusarium wilt of chickpea. A marked increase in shoot and root length was observed in P. fluorescens treated plants. The isolates of P. fluorescens systemically induced resistance against Fusarium wilt of chickpea caused by Fusarium. oxysporum f.sp. ciceri (FocRs1), and significantly (P = 0.05) reduced the wilt disease by 26-50% as compared to control. Varied degree of protection against Fusarium wilt was recorded with chemical inducers. The reduction in disease was more pronounced when chemical inducers were applied with P. fluorescens. Among chemical inducers, SA showed the highest protection of chickpea seedlings against wilting. Fifty two- to 64% reduction of wilting was observed in soil treated with isolate Pf4-92 along with chemical inducers. A significant (P = 0.05; r = -0.946) negative correlation was observed in concentration of salicylic acid and mycelial growth of FocRs1 and at a concentration of 2000 microg ml(-1) mycelial growth was completely arrested. Exogenously supplied SA also stimulated systemic resistance against wilt and reduced the disease severity by 23% and 43% in the plants treated with 40 and 80 microg ml(-1) of SA through root application. All the isolates of P. fluorescens produced SA in synthetic medium and in root tissues. HPLC analysis indicated that Pf4-92 produced comparatively more SA than the other isolates. 1700 to 2000 nanog SA g(-1) fresh root was detected from the application site of root after one day of bacterization whereas, the amount of SA at distant site ranged between 400-500 nanog. After three days of bacterization the SA level decreased and was found more or less equal at both the detection sites. PMID- 14521231 TI - In silico analysis of a flavohemoglobin from Sinorhizobium meliloti strain 1021. AB - Hemoglobins (Hbs) have been characterized from a wide variety of eubacteria, but not from nitrogen-fixing rhizobia. Our search for Hb-like sequences in the Sinorhizobium meliloti genome revealed that a gene coding for a flavohemoglobin (fHb) exists in S. meliloti (SmfHb). Computer analysis showed that SmfHb and Alcaligenes eutrophus fHb are highly similar and could fold into the same tertiary structure. A FNR-like box was detected upstream of the smfhb gene and mapping analysis revealed that the smfhb gene is flanked by nos and fix genes. These observations suggest that smjhb is regulated by the concentration of O2 and that SmfHb functions in some aspects of nitrogen metabolism. PMID- 14521232 TI - Differential amplification of sequence heterogeneous ribosomal RNA genes and classification of the 'Fragaria multicipita' phytoplasma. AB - Ribosomal (r) RNA interoperon sequence heterogeneity in the 'Fragaria multicipita' phytoplasma, a member of group 16SrVI, was initially observed in RFLP patterns of rDNA amplified in the polymerase chain reaction (PCR), and was confirmed through sequence analysis of cloned rDNA. Sequences from operons rrnA and rrnB were amplified in PCR primed by primer pair P1/P7 but from only rrnA in PCR primed by primer pair R16mF2/R16mR1. Preferential amplification of DNA from operon rrnA was explained by base mismatches between the R16mF2/R16mR1 primers and primer annealing sites in rrnB. The results revealed potential for classification of a phytoplasma into two different subgroups within a 16S rRNA group, if the phytoplasma's 16S rRNA gene sequences are independently characterized. It is suggested that the rRNA operon containing species-specific signature sequence(s) should be specified, and where possible sequences from both 16S rRNA genes should be included, in descriptions of new 'Candidatus Phytoplasma species'. PMID- 14521233 TI - Purification and characterization of an aflatoxin degradation enzyme from Pleurotus ostreatus. AB - Nineteen fungi were tested for their ability to degrade aflatoxin B1 (AFB1). An extracellular enzyme from the edible mushroom Pleurotus ostreatus showed afaltoxin-degradation activity detected by thin-layer chromatography (TLC). An enzyme with this activity was purified by two chromatographies on DEAE-Sepharose and Phenyl-Sepharose. The apparent molecular mass of the purified enzyme was estimated to be 90 kDa by SDS-PAGE. Optimum activities were found in the pH range between 4.0 and 5.0 and at 25 degrees C. Also, degradation activity of several dyes in the presence of H2O2 was tested, resulting in the detection of bromophenol blue-decolorizing activity. Based on these data, we suggest this enzyme is a novel enzyme with aflatoxin-degradation activity. Fluorescence measurements suggest that the enzyme cleaves the lactone ring of aflatoxin. PMID- 14521234 TI - Growth promoting influence of siderophore-producing Pseudomonas strains GRP3A and PRS9 in maize (Zea mays L.) under iron limiting conditions. AB - Maize seeds were bacterized with siderophore-producing pseudomonads with the goal to develop a system suitable for better iron uptake under iron-stressed conditions. Siderophore production was compared in fluorescent Pseudomonas spp. GRP3A, PRS9 and P. chlororaphis ATCC 9446 in standard succinate (SSM) and citrate (SCM) media. Succinate was better suited for siderophore production, however, deferration of media resulted in increased siderophore production in all the strains. Maximum siderophore level (216.23 microg/ml) was observed in strain PRS9 in deferrated SSM after 72 h of incubation. Strains GRP3A and PRS9 were used for plant growth promotion experiments. Strains GRP3A and PRS9 were also antagonistic against the phytopathogens, Colletotrichum dematium, Rhizoctonia solani and Sclerotium rolfsii. Bacterization of maize seeds with strains GRP3A and PRS9 showed significant increase in germination percentage and plant growth. Maximum shoot and root length and dry weight were observed with 10 microM Fe3+ along with bacterial inoculants suggesting application of siderophore producing plant growth promoting rhizobacterial strains in crop productivity in calcareous soil system. PMID- 14521235 TI - Molecular characterization of Macrophomina phaseolina and Fusarium species by a single primer RAPD technique. AB - Charcoal root rot and wilt, are two economically important diseases of many crop plants in North and South America, Asia and Africa and some parts of Europe. Genetic variation in 43 isolates of Macrophomina phaseolina and 22 isolates of Fusarium species, collected from geographically distinct regions over a range of hosts, was studied using random amplified polymorphic DNA (RAPD) markers. Initially, 210 arbitrary nucleotide (10-mer) primers were tested for amplification of genomic DNA of one M. phaseolina isolate, 70 primers amplified the genomic DNA of M. phaseolina. One primer OPA-13 (5'-CAGCACCCAC-3') produced fingerprint profiles, which clearly distinguished between the different isolates of M. phaseolina. UPGMA analysis classified these isolates into five major groups. By primer OPA-13, 22 isolates of pathogenic and non-pathogenic Fusarium species of different formae-speciales and races, were also distinguished from M. phaseolina. This marker is useful for distinguishing between these two important plant pathogens irrespective of hosts, virulence spectrum and races. This is the first report of reliable diagnosis of two soilborne pathogens (root/collar rot and wilt causing pathogens) at the level of isolates, formae-speciales and races by a single primer RAPD procedure with uniform PCR conditions. PMID- 14521236 TI - Antimicrobial potentiality of a new non-antibiotic: the cardiovascular drug oxyfedrine hydrochloride. AB - Ten cardiovascular drugs, having diverse pharmacological action, were screened for possible antimicrobial property against known eight sensitive bacteria, belonging to Gram positive and Gram negative types. Although five drugs failed to show antimicrobial activity and three had moderate antimicrobial action, oxyfedrine HCl and dobutamine were seen to possess pronounced antimicrobial property. Oxyfedrine was further tested in vitro against 471 strains of bacteria from two Gram positive and fourteen Gram negative genera. The minimum inhibitory concentration (MIC) of oxyfedrine was determined by agar dilution method, which ranged from 50-200 microg/ml in most of the strains, while some strains were inhibited at even lower concentrations. In animal experiments, this compound was capable of offering significant protection to Swiss strain of white mice, challenged with 50 median lethal dose (MLD) of a virulent strain of Salmonella typhimurium at concentrations of 15, 30 and 60 microg/mouse. The in vivo results were highly significant according to chi-square test. PMID- 14521238 TI - Youth tobacco cessation: filling the gap between what we do and what we know. PMID- 14521237 TI - Interaction between gfp-tagged Pseudomonas tolaasii P12 and Pleurotus eryngii. AB - Pseudomonas sp., (formerly reported as strain P12) which produces brown blotch disease symptoms on Pleurotus eryngii, has been identified as P. tolaasii based on its biochemical, physiological properties and 16S rDNA sequence analysis. This pathogen is able to infect basidiocarps when surface-inoculated on mushroom casing soil. However, infected basidiocarps develop the brown blotch disease symptoms when the pathogen concentration in the fruiting body tissues is higher than 10(4) cfu/g d.w. Using gfp-tagged cells and confocal laser scanning microscopy, it was possible to show that the pathogen has the ability to tightly attach to the hyphae of Pleurotus eryngii. PMID- 14521239 TI - Youth Tobacco Cessation Collaborative and National Blueprint for Action. AB - OBJECTIVES: To describe the formation of the Youth Tobacco Cessation Collaborative (YTCC), a voluntary collaborative of leading funders of youth tobacco cessation research and services. METHODS: The long-term goal and specific short-term (2-year) goals, strategies, and accomplishments are briefly described with reference to its guiding action plan: National Blueprint for Action: Youth and Young Adult Tobacco-Use Cessation. RESULTS: Aiming to accelerate the pace of discovery and application, YTCC efforts have created a strategic vision for making progress toward filling key knowledge and intervention gaps. CONCLUSIONS: Lessons learned about effective partnership are reviewed, and future directions are described. PMID- 14521240 TI - A comparison of review methods in tobacco prevention and control guidelines. AB - OBJECTIVES: To compare methods and conclusions of selected guidelines evaluating youth tobacco interventions. METHODS: Eight reviews were compared regarding (a) search and assessment of evidence, (b) translation into recommendations, and (c) conclusions regarding tobacco cessation among youth. RESULTS: Narrative and systematic review methods were employed. Five systematic reviews adopted different (a) criteria for study inclusion, (b) methods estimating summary outcome measurements, and (c) standards for concluding on the evidence of effectiveness. CONCLUSIONS: Evidence reviews provide useful summaries, conclusions, and recommendations. Youth Tobacco Cessation: A Guide for Making Informed Decisions provides an inital systematic review of the evidence on effectiveness of youth tobacco use. PMID- 14521241 TI - Application of a better practices framework to review youth tobacco use cessation. AB - OBJECTIVE: To describe the Better Practices model and its application to review youth tobacco cessation practices. METHODS: Growing emphasis on evidence-based practice in health has drawn attention to methods for the identification of "best practices." Canadian organizations and government collaborated to develop a framework for "better" practices. RESULTS: An international group of experts used the model to develop recommendations for the practice of youth cessation and further research needed to advance the field. CONCLUSIONS: Tension persits between the need for rigor in research and the need for practical intervention guidance. The Better Practices model provided a framework for action recognizing these needs, and its use resulted in a number of valuable products. PMID- 14521242 TI - Better practices for youth tobacco cessation: evidence of review panel. AB - OBJECTIVES: To offer programmers, policy makers, and researchers a scientific basis for developing and selecting smoking cessation treatments for adolescents. METHODS: An evidence review panel systematically rated published and unpublished reports of cessation treatments for youth to make recommendations on theoretical foundations, delivery settings, types of intervention, and provider type. RESULTS: Twenty studies had sufficient validity to inform the recommendations. The 9 studies that reported treatments that increased cessation were based on social cognitive theory. CONCLUSIONS: Cognitive-behavioral interventions are a promising approach for helping young smokers quit smoking. Evidence is insufficient to draw other conclusions at this time. PMID- 14521243 TI - Recommendations and guidance for practice in youth tobacco cessation. AB - OBJECTIVE: To summarize recommendations from Youth Tobacco Cessation: A Guide for Making Informed Decisions for careful consideration, selection, implementation, and evaluation of youth cessation interventions. METHODS: Recommendations were developed from an evidence review and consensus from a multidisciplinary advisory panel. RESULTS: Identified essential elements for selecting, planning, delivering, and evaluating youth cessation interventions. CONCLUSIONS: Until there is more evidence for effectiveness of youth specific cessation interventions, clinicians and practitoners should adopt treatments that use cognitive-behavioral approaches for youth cessation interventions that require careful planning and rigorous evaluation. PMID- 14521244 TI - Improving the future of youth smoking cessation. AB - OBJECTIVES: To provide recommendations that will build a better foundation for research on youth smoking cessation. METHODS: The Youth Tobacco Collaborative Cessation panel evaluated youth tobacco cessation literature and convened meetings to reach consensus. RESULTS: Methodological issues include design, recruitment and retention, follow-up, measurement, and youth vernacular. Research gaps include youth characteristics, theoretical approaches, delivery settings, and type of provider. Thirteen key research components for reporting are addressed. CONCLUSIONS: Given the dearth of studies on youth smoking cessation, scientifically rigorous studies need to be conducted with attention to methodological issues, research gaps, and reporting of key research components. PMID- 14521245 TI - Coping with youth violence: assessments by minority parents in public housing. AB - OBJECTIVE: To present the views of predominantly African American public housing residents as they discuss violence prevention. METHODS: Qualitative research was conducted with 38 parents of adolescents. Data were analyzed inductively and in relation to an appraisal-coping theoretical framework. RESULTS: Parents enumerated cues that signaled neighborhood danger and signs and symptoms that suggested youth distress. Narratives reflect both ad hoc and systematic coping responses to incidences of violence that involve their children. CONCLUSION: Parents used a variety of coping strategies and diverse resources such as personal skills, family strengths, and community agencies in an attempt to decrease youth exposure and involvement in violence. PMID- 14521246 TI - Social support and social norms: do both contribute to predicting leisure-time exercise? AB - OBJECTIVE: To clarify the contribution of social support and social norms to exercise behavior. METHODS: A sample of 363 college students completed a questionnaire that assessed social support and social negativity from friends, descriptive and injunctive social norms related to friends, perceived behavioral control, attitude, intention, and leisure-time exercise. RESULTS: Esteem social support was the strongest predictor of total and strenuous leisure-time exercise (P < .001), and descriptive norm was a significant (P < .01 predictor of strenuous leisure-time exercise. CONCLUSION: Social support and social norms contribute independently to our understanding of variation in the frequency of strenuous leisure-time exercise. PMID- 14521247 TI - Developing theoretical and environmental policy for underage drinking. AB - OBJECTIVE: To estimate concern/support for policy for underage drinkers and formulate policy-specific communication strategies. METHODS: An environmentally based approach to policy formulation used concepts from the transtheoretical, decision-balance, and social-norms models. Interviews were conducted with community stakeholders, and surveys were collected on community residents as part of the evaluation effort. RESULTS: The triangulated evaluation revealed specific policies for underage drinking and techniques for facilitating policy implementation. CONCLUSION: The results demonstrate techniques for using multiple models and evaluation techniques for formulating public policy in the promotion of community health. PMID- 14521248 TI - An intervention to promote walking in sedentary women in the community. AB - OBJECTIVE: To evaluate a telephone counseling intervention that was designed to help sedentary women begin and maintain a walking program. METHODS: Females (N = 197) were randomly assigned to either an intervention, attention control, or no attention control group. Assessments were made at baseline and 6 months. RESULTS: Women in the intervention group reported more time walked each day than did control women (P < .05). The intervention worked equally for African American and European American women as well as for different income groups. CONCLUSION: Overall, a counseling intervention via telephone appears to be a good way to help women begin a walking program. PMID- 14521249 TI - A su salud: a quasi-experimental study among Mexican American women. AB - OBJECTIVES: To test the effectiveness of a community intervention program to promote breast and cervical cancer screening. METHOD: Low-income Mexican American neighborhoods located in 2 Texas cities were assigned to intervention or comparison conditions. A 2-year community intervention was delivered via local mass media and reinforced by indigenous volunteers. RESULTS: There were higher Pap smear completion rates for women under 40 years of age in the intervention community. CONCLUSION: Although it is important to address the cultural needs of all Mexican American women, it is also important to understand the tangible environmental barriers faced by the older women. PMID- 14521250 TI - Prevalence and correlates of depressive symptoms in young adolescents. AB - OBJECTIVES: To assess prevalence of elevated depressive symptoms in young adolescents and examine associations between symptoms and sociodemographic and behavioral factors. METHODS: Cross-sectional survey data from 3621 seventh grade students from 16 middle schools were analyzed. RESULTS: Elevated depressive symptoms were reported by 40% of girls and 30% of boys. Socioeconomic status, race/ethnicity, and age group were independently associated with depressive symptomatology. For girls, monthly alcohol use, monthly smoking, heavy drinking, and inhalant use were significant correlates. For boys, monthly alcohol use and inhalant use were significant. CONCLUSIONS: Elevated depressive symptomatology was a prevalent problem. Substance use was often associated with depressive symptoms, especially among girls. PMID- 14521251 TI - Smokers' willingness to protect children from secondhand smoke. AB - OBJECTIVE: To examine the effectiveness of a secondhand smoke media campaign on adult smokers' willingness to protect children from secondhand smoke. METHODS: Following a series of community awareness ads, a random sample of 390 adult smokers was surveyed via telephone regarding their perceptions of secondhand smoke. RESULTS: Seeing or hearing the ads was significantly associated with intentions to personally smoke outside of the home and to ask visitors to smoke outside. Most smokers were committed to protecting children from secondhand smoke. CONCLUSION: Secondhand-smoke media campaigns can positively affect smokers' knowledge of the dangers of secondhand smoke and future intentions to protect others from secondhand smoke. PMID- 14521252 TI - How renowned researchers were initiated into the world of research. PMID- 14521253 TI - Comparison of propofol and sevoflurane for laryngeal mask airway insertion. AB - In this study, we compared haemodynamic changes, laryngeal mask airway (LMA) insertion time, and complications in patients anesthetized by inhalation of sevoflurane with those of intravenous induction with propofol. One hundred patients, aged between 20-40 years were enrolled in this study. Group 1 received propofol (2.5 mg/kg i.v.) and group 2 received sevoflurane (6%+50% N2O+50% O2) by inhalation using the tidal volume technique. LMA insertion time was found to be significantly longer in sevoflurane group than in propofol group. Mean arterial blood pressure was significantly lower within each group after induction in comparison to before induction values. In both groups, the LMA was successfully inserted in all patients. The quality of anaesthesia according to patients was significantly higher in the propofol group (80%) than in sevoflurane group (30%). Odor perception was significantly higher in sevoflurane group (84%) than in propofol group (38%). Apnoea was significantly higher in propofol group (40%) than in sevoflurane group (0%). Sevoflurane is an alternative to propofol for induction of anaesthesia and has a lower incidence of apnoea. Other complication rates are not higher than which propofol but the longer duration of induction time is a disadvantage. PMID- 14521255 TI - Effects of contralateral noise on the measurement of auditory threshold. AB - It is well known that sound presented in the contralateral ear can elicit the activity of the olivocochlear (OC) efferent. In the present study, the effects of the addition of contralateral noise on the psychophysical measurements of auditory thresholds were investigated in human subjects with normal hearing. The results obtained in the present study indicate that the addition of contralateral noise at a level of only 20 or 30 dB sound pressure level (SPL) may cause a significant elevation of the auditory threshold in the mid-frequency area (usually 2-3 dB). When the level of contralateral noise was elevated, the elevation of the auditory threshold tended to be larger and the affected frequency area became wider. Although other factors that elevate the auditory thresholds, such as cross-talk effects and the acoustic reflex of the middle ear muscles, may be involved in the above-mentioned paradigm, especially when higher levels of contralateral noise are used, it is important to know the degree of OC mediated threshold elevation in usual audiometric measurement. PMID- 14521254 TI - Comparative assessment of chordal preservation versus chordal resection in mitral valve replacement for mitral stenosis (long-term follow-up: 8 years). AB - Carrying out a mitral valve replacement (MVR) while preserving all chordae tendineae in patients with mitral regurgitation has been proven beneficial to the left ventricular performance. To evaluate the effectiveness of this technique in patients with mitral stenosis, a comparison of the echocardiographic data between patients who were operated on using this technique (Group Preservation = GroupP, n = 15), and those operated on using the conventional method of MVR (Group Conventional = GroupC, n = 15) was made. All patients were examined before surgery, 6 months after surgery and 8 years after surgery. The study population was limited to patients who had no evidence of coronary artery disease, aortic stenosis and/or regurgitation and patients who had pure mitral stenosis. Echocardiographic measurements obtained both 6 months and 8 years postoperatively revealed a significant decrease in the left ventricular ejection fraction in GroupC (61.33+/-9.29% preoperatively, 53.2+/-10.3% postoperatively). The difference between the decrease in the left ventricular ejection fraction of the two groups was statistically significant (-0.71+/-6.28% in GroupP, -8.07+/-13.35% in GroupC). There was no evidence of prosthetic valve dysfunction and no operative deaths. Two patients died in GroupP, 3 patients died in GroupC within the 8 year period. The conclusion was reached, that if suitable, mitral valve replacement while preserving the chordae tendineae is expected to have a beneficial effect on postoperative left ventricular performance in patients with mitral stenosis. PMID- 14521256 TI - Expression of glutamate transporter GLAST in the developing mouse cochlea. AB - The immunohistochemical localization of glutamate transporter GLAST in the developing mouse cochlea was studied at different ages between 0 and 30 days after birth (DAB). In the adult mouse cochlea, intense GLAST-like immunoreactivity was found in the supporting cells adjacent to the inner hair cells of the organ of Corti, the type II and suprastrial fibrocytes of the cochlear lateral wall, the fibrocytes of the spiral limbus and the satellite cells surrounding the spiral ganglion cells. At 0 DAB, weak GLAST-like immunoreactivity was found in the supporting cells around the immature inner hair cells. Immature fibrocytes in the cochlea were also positively immunostained. At 3 DAB, weak immunostaining of GLAST appeared in the immature satellite cells in the spiral ganglion. The GLAST-like immunoreactivity in the supporting cells around the inner hair cells, in the fiborocytes in the spiral ligament and the spiral limbus and in the satellite cells in the spiral ganglion increased progressively during the second postnatal week, and reached the adult level at 15 DAB. This time course correlates with the electrophysiological onset and maturation of the mouse auditory function, which is mediated by glutamatergic neurotransmission. These results suggest that the expression of GLAST may be needed for the efficient removal and metabolism of the released glutamate in the cochlea and may play important roles in the onset and maturation of the auditory system. PMID- 14521257 TI - Successful pregnancy after myomectomy using preoperative adjuvant uterine artery embolization. AB - A 30-year-old-woman presented with uterine leiomyoma and primary sterility. Abdominal myomectomy was performed; however, one cervical leiomyoma was not resected because of a risk of excessive blood loss. Two years after the procedure, a secondary myomectomy using preoperative adjuvant uterine artery embolization (UAE) was performed because of pronounced menorrhagia and her hope for bearing children. The patient's dysmenorrhea disappeared postoperatively and she conceived spontaneously 3 years after the secondary myomectomy. This case suggests that myomectomy using preadjuvant UAE may be an another approach for the treatment of leiomyoma in patients who wish bear children in the future. PMID- 14521258 TI - End-stage kidney at the onset of nephrotic syndrome in a 4-year-old girl. AB - A previously healthy Japanese girl aged 4 years suddenly developed idiopathic nephrotic syndrome (INS) associated with acute renal failure (ARF). Despite prompt initiation of hemodialysis and prednisolone treatment, her ARF did not recover. A percutaneous renal biopsy performed within a month from the onset of INS revealed lesions of end-stage kidney, which is unusual for the acute stage of INS. Although the etiology of her ARF remains speculative, an acute exacerbation of an underlying chronic disease, such as non-nephrotic focal segmental glomerulosclerosis might be the cause. PMID- 14521259 TI - Polymorphisms of heme oxygenase-1 and bilirubin UDP-glucuronosyltransferase genes are not associated with Kawasaki disease susceptibility. AB - Kawasaki disease (KD) is a systemic vasculitis and occurs among Japanese children at a high incidence. Serum bilirubin and heme oxygenase-1 (HO-1) expression are known to play a significant role in the protection of vascular endothelial cells. Japanese have unique polymorphic distribution patterns of (TA)7 or G71R of the bilirubin UDP-glucuronosyltransferase (B-UGT) gene and of (GT)n repeats of the HO 1 gene. We investigated the relationship of KD susceptibility with these polymorphisms. There were no significant differences in the distribution of allele frequencies and genotypes of these polymorphisms between KD patients and controls. These polymorphisms are not associated with KD susceptibility. PMID- 14521260 TI - Antiviral activity of 2-amino-4,4alpha-dihydro-4alpha-7-dimethyl-3H-phenoxazine-3 one on poliovirus. AB - 2-Amino-4,4alpha-dihydro-4alpha-7-dimethyl-3H-phenoxazine-3-one (Phx), which was produced by the reaction of bovine hemoglobin with 2-amino-5-methylphenol, inhibited the proliferation of poliovirus in Vero cells between 0.25 microg/ml and 2 microg/ml with maximal antiviral acitivity at 1 microg/ml. These results suggest that Phx may be useful to prevent the proliferation of poliovirus infection. PMID- 14521261 TI - Relative quantification of ERBB2 mRNA in invasive duct carcinoma of the breast: correlation with ERBB-2 protein expression and ERBB2 gene copy number. AB - The option to treat patients suffering from ERBB-2 protein-positive invasive duct carcinomas of the breast (IDC) with Herceptin requires a precise determination of the ERBB2 status. The aim of the study was to evaluate the ERBB2 mRNA level, placing emphasis on cases with discordant findings between ERBB-2 protein expression (IHC) and a copy number of the ERBB2 gene (FISH). Thirty-nine IDCs (21 cases IHC and FISH concordant, 15 cases moderately discordant, 3 cases markedly discordant) were investigated. ERBB2 mRNA expression was determined using quantitative real-time RT-PCR (Q-RT-PCR). IDCs with negative ERBB-2 protein and without ERBB2 gene amplification had a low ERBB2 mRNA level. Cases with 3+ overexpression of the protein and with strong gene amplification (> 10 copies/tumor cell) had a significantly increased expression of ERBB2 mRNA. In 13 of 15 IDCs with moderate discrepancies (up to 10 copies of the gene per one tumor cell/negative ERBB-2 protein; without amplification/2+ protein) mRNA was low, comparable to that in cases with negative ERBB-2 protein and without ERBB2 gene amplification. In three cases with markedly discordant findings (the gene amplified/protein negative--one case; protein 3+/no amplification--2 cases), Q-RT PCR results were within a "normal" limit. Ineffective gene amplification and protein accumulation are suggested explanations. Q-RT-PCR revealed two cases with highly expressed ERBB2 mRNA and discordant FISH and/or IHC findings. Increased effectiveness of transcription (protein 2+/high mRNA/without the gene amplification), and combined dysregulation (protein negative/high mRNA/no amplification) are possible causes of these findings. Q-RT-PCR appears useful in clarifying borderline or discrepant IHC and FISH findings. PMID- 14521262 TI - Neuronal nuclear antigen (NeuN): a new tool in the diagnosis of central neurocytoma. AB - The use of neuronal nuclear antigen (NeuN) as a reliable neuronal marker in the differential diagnosis of clear cell neoplasms of the central nervous system was determined in a biopsy series of 23 cases. Immunohistochemical analyses were carried out by antisera against neuronal nuclear antigen, synaptophysin, neuron specific enolase, microtubule-associated protein 2, and glial fibrillary acidic protein. All eight central neurocytomas were characteristically immunolabeled by NeuN. NeuN immunoreactivity was uniformly strong and basically located in the nuclei of neurocytes. Despite this uniform staining pattern of central neurocytomas, 12 cases of oligodendrogliomas and three cases of ependymoma were negative for NeuN. As the diagnostic criteria for central neurocytoma include immunohistochemical and/or ultrastructural evidence for neuronal differentiation, NeuN as a sensitive and specific neuronal marker in formalin-fixed, paraffin embedded tissues may greatly facilitate the differential diagnosis of central neurocytomas. PMID- 14521263 TI - Vascularization pattern in hypertrophic scars and keloids: a stereological analysis. AB - Wound healing is a complex process that does not always occur harmoniously and may lead to pathological scar development, such as hypertrophic scars and keloids. Considering that vascularization can play a role in the development of these scars, and that the literature is controversial, we performed a stereological analysis of dermal for vessels of normal skin, normal scars, hypertrophic scars, and keloids. The parameters studied concerned vessels: surface density, length density; for vessels and myofibroblasts: volume density, in papillary and reticular dermis. The pattern of dermal vascularization in normal skin and normal scar showed no differences. In papillary demis, the number of vessels was higher in hypertrophic scars and keloids than in normal skin (p < 0.05). Vessels of hypertrophic scars were more dilated than those of normal skin (p < 0.01). In reticular dermis, vessels were present in higher amount in hypertrophic scars and keloids than in normal skin (p < 0.025; p < 0.001, respectively). The pattern of vascularization did not show any differences between hypertrophic scars and keloids. Our results show that hypertrophic scars and keloids have a distinct pattern of vascularization compared to normal skin and normal scars. This indicates that abnormal vascularization can be involved in the development of hypertrophic scars and keloids. PMID- 14521264 TI - Quantification of the global and local complexity of the epithelial-connective tissue interface of normal, dysplastic, and neoplastic oral mucosae using digital imaging. AB - This study aimed at quantifying the complexity of the epithelial-connective tissue interface (ECTI) in human normal mucosa, premalignant, and malignant lesions using fractal geometry. Two approaches were used to describe the complexity of 377 oral mucosa ECTI profiles. The box counting method was used to estimate their global fractal dimension, while local fractal dimensions were estimated using the mass radius relation at various local scales. The ECTI complexity significantly increased from normal through premalignant to malignant profiles in both global and local (over 283 microm) scales. Normal mucosa samples from different sites of the oral cavity also had different degrees of global complexity. Fractal geometry is a useful morphological marker of tissue complexity changes taking place during epithelial malignancy and premalignancy, and we propose it as a quantitative marker of epithelial complexity. PMID- 14521265 TI - Bone marrow infiltrate by a poorly differentiated neuroendocrine carcinoma. AB - We report a rare case of bone marrow infiltrate by a neuroendocrine carcinoma. The patient was a 66-year-old man who presented with anemia and thrombocytopenia and with a leukoerythroblastic picture on blood films. A bone marrow biopsy revealed extensive tumor infiltrate with morphological and immunohistochemical features of neuroendocrine origin. There were also multiple nodular lesions in the liver, which showed the same pathology as that in the bone marrow. Since none of the lesions could be identified as the primary tumor, the patient was diagnosed to have a poorly differentiated neuroendocrine carcinoma of unknown primary site. PMID- 14521266 TI - Carcinosarcoma and squamous cell carcinoma of the renal pelvis associated with nephrolithiasis: a case report of each tumor type. AB - Both squamous cell carcinoma and carcinosarcoma of the renal pelvis are uncommon. We report on two cases, one carcinosarcoma and one squamous cell carcinoma of the renal pelvis. In the patient with squamous cell carcinoma of the left kidney, the tumor was neither detectable on preoperative radiological evaluation nor grossly visible in the surgical specimen. This patient, a 56-year-old man, presented with left lumbar pain, hematuria, fever, 4-5 kg weight loss, and untreated nephrolithiasis of the left kidney that had been diagnosed 20 years earlier. The second patient, an 87-year-old woman, also had a long history of left kidney nephrolithiasis and presented with left lumbar pain and hematuria. Both patients underwent nephrectomy for removal of the non-functioning hydronephrotic left kidney. In both cases, microscopic examination of the surgical specimen revealed squamous metaplasia and dysplasia in the pelvicalyceal mucosa, and islands of atypical squamous cells in the renal parenchyma. In the second case, the kidney also showed sarcomatous changes in the pelvis. PMID- 14521267 TI - Pseudoglandular myxoid adenoma of the adrenal gland. AB - Myxoid tumors of the adrenal cortex are rare. To the best of our knowledge, only 17 cases have been reported to date, including 10 carcinomas and 7 adenomas. The myxoid areas of these tumors are often evident grossly, and their extent is variable. We report on a case of a myxoid adenoma of the right adrenal gland in a 45-year-old male with a prominent pseudoglandular arrangement in more than 90% of the tumor mass, and with a minor component represented by anastomosing cords of tumor cells floating in pools of myxoid material. In addition, after extensive examination, we found foci of typical adrenocortical adenoma. Grossly, the tumor was yellowish without discernable gelatinous changes. Most of the tumor cells had a moderate amount of clear vacuolated cytoplasm and contained numerous oil-red-O positive lipid droplets. Extracellularly, in the lumens of pseudoglands and on the background, we noticed myxoid material that stained positively with Alcian blue (pH 2.5) and weakly positively with mucicarmine and the PAS method. Immunohistochemical examination showed positivity of the tumor cells for vimentin and cytokeratin CAM5.2. Synaptophysin was weakly positive only focally. Cytokeratin AE1/AE3, EMA, and CEA were negative. PMID- 14521268 TI - Riedel's thyroiditis containing cytologically atypically appearing B-cells: a case report. AB - We report on a case of Riedel's thyroiditis containing cytologically atypically appearing B-cells. The patient was a 60-year-old Japanese female, who had a well demarcated tumor that showed extensive growth from the right thyroid lobe into perithyroidal soft tissues. Histologically, the lesion consisted of two components. The central tumor-like portion of the mass showed spindle cells intermixed with abundant collagen fibers. In addition, scattered B-cells with large nuclei and multiple small, but conspicuous nucleoli were also present. At the periphery of the nodule, numerous plasma cells and small lymphocytes, as well as occasional B-immunoblasts and plasmablasts were growing between non-neoplastic thyroid follicles. An infiltration of thyroid follicles by B-cells also resulted in lymphoepithelial-like lesions. The specimen obtained from an enlarged cervical lymph node occurring 14 months later contained numerous mature plasma cells showing a sheet-like arrangement with occasional immature plasma cells and immunoblasts in the interfollicular area and medullary cords. The polyclonal nature of B-cells was demonstrated immunohistochemically and by polymerase chain reaction in both the thyroid gland and the lymph node lesion. PMID- 14521269 TI - Intermuscular chondrolipoma of the thigh: the diagnostic way of a rare entity. AB - Cartilaginous metaplasia in lipomas (chondrolipoma) is rare and mainly encountered in large-sized, long-standing lipomas. Chondrolipomas can be found at almost any site of the body, particularly in the connecting tissue of the skeletal system, breast, pharynx, and nasopharynx. We report on an intermuscular tumor of the thigh in a patient who suffered from lipomatosis in his past medical history. We describe how the diagnosis of chondrolipoma was reached and discuss the differential diagnoses. PMID- 14521270 TI - Severe cytological atypia (large cell change) in focal nodular hyperplasia with numerous mallory bodies. A benign (adaptive) change? AB - Focal nodular hyperplasia (FNH) is a benign hepatocellular lesion composed of hyperplastic appearing hepatocytes arranged in nodules separated by fibrous septa that usually form a central stellate scar. Rare lesions that show unusual cytological or architectural features were reported as variants of focal nodular hyperplasia. We present the morphological features of a case of FNH with severe cytological atypia (so-called large cell change) in a 73-year-old man. In addition to diffuse cytological atypia, Mallory hyaline bodies were found in almost all lesional cells. This rare variant of FNH should be differentiated from other neoplastic lesions, in particular from the fibrolamellar variant of hepatocellular carcinoma. PMID- 14521271 TI - The clinical relevance of EEG interpretation. AB - There is need nowadays to re-emphasize the capabilities of electroencephalography: a method representing the extremely important function/dysfunction-orientation in neurological thinking and practice. Valuable and relevant messages to the clinician naturally require solid EEG training and the resulting expertise. The idea that valuable EEG information is limited to the field of epileptology is erroneous. A plethora of clinically relevant messages can be derived from the EEG in nonepileptic conditions and, above all, in metabolic (and so called "mixed") encephalopathies where neuroimaging has almost nothing to offer. The discussion of EEG and epileptology only skirts pediatric conditions (and most of the epileptic syndromes). It is shown that EEG reading in epileptology is a lot more than simply "hunting spikes." A strong plea is being made against the presently fashionable overuse of the term "non-convulsive status epilepticus." Continuing neglect of functional/dysfunctional orientation can seriously endanger the entire field of Neurology. PMID- 14521272 TI - Event-related correlations in learning impaired children during A hybrid go/no-go choice reaction visual-motor task. AB - One hundred sixty-nine learning impaired (LI) and 71 non-learning impaired (NLI) children underwent a hybrid go/no-go choice reaction time visual-motor task to study the behavioral and physiological fundamentals of learning disorders. A left button was pressed for Left Arrow (LA) stimuli, a right for Right Arrow (RA) stimuli, none (no-go) for a non-directional arrow. Stimulus specific visual evoked potentials were formed and, with PZ as index electrode, were lag correlated to frontal electrodes to form Event-Related Correlations (ERC). Exploratory t-statistic significance probability maps (t-SPM) were used to define regions of interest (ROI). Behaviorally, there was a right-hand advantage over the left in the NLI group, but less in the LI group. Electrophysiologically, RA and LA conditions increased correlation between visual areas (PZ) and contralateral frontal areas (F3 and F4). A unilateral ROI, at electrode FC1, also preceded both left- and right-handed responses. Neurobehaviorally, increased visual-motor correlation was associated with better performance, especially for the left hemisphere, at F3 and FC1. Surprisingly, visual-motor correlations were not associated with performance for the NLI group in the RA and no-go condition. Our data support previously reported difficulties of learning impaired children in low-level information processing. Furthermore, we hypothesize that LI, in contrast to NLI children, demonstrate difficulty in automatizing routine tasks. PMID- 14521273 TI - Quantitative electroencephalographic studies of cue-induced cocaine craving. AB - Quantitative electroencephalographic (qEEG) profiles were studied in cocaine dependent patients in response to cocaine cue exposure. Using neurometric analytical methods, the spectral power of each primary bandwidth was computed and topographically mapped. Additional measures of cue-reactivity included cocaine craving, anxiety and related subjective ratings, and physiological measures of skin conductance, skin temperature, heart rate, and plasma cortisol and HVA levels. Twenty-four crack cocaine-dependent subjects were tested for their response to tactile, visual and audio cues related to crack cocaine or neutral items. All measures were analyzed for significant difference by comparing cocaine versus neutral cue conditions. An increase in cocaine craving, anxiety and related subjective ratings, elevated plasma cortisol levels, and a decrease in skin temperature, were induced by cocaine cue exposure. Distinct qEEG profiles were found during the paraphernalia handling and video viewing (eyes-open), and guided imagery (eyes-closed), phases of cocaine cue exposure. During paraphernalia handling and video viewing, there was an increase in beta activity accompanied by a drop in delta power in the frontal cortex, and an increase in beta mean frequency in the occipital cortex. In contrast, during guided imagery there was an increase in theta and delta power in the frontal cortex, and an increase in beta power in the occipital cortex. Correlation analyses revealed that cue-induced anxiety during paraphernalia handling and video viewing was associated with reduced high frequency and enhanced low frequency EEG activity. These findings demonstrated that EEG activation during cue-induced cocaine craving may be topographically mapped and subsequently analyzed for functional relevance. PMID- 14521274 TI - P300 (latency) event-related potential: an accurate predictor of memory impairment. AB - To determine if P300 latency changes precede and correlate with memory and mental status, patients (N=1506 aged 20-100 years) who received medical and psychiatric diagnoses (from 1997 to 2002), were assessed for P300 (N=1496), WMS-III (N=694), and MMSE (N=456). Patient and control groups included, a) normal WMS-III on all 4 subscales (N=36), b) normal WMS-III and MMSE (N=189) with subjective memory/mental status complaints, and c) medical patients with normal WMS-III and no memory complaints (N=205), and d) P300 control group without medical, psychiatric or memory problems for ROC. Patients with impaired/borderline memory had a prolonged P300 latency (P<0.02) compared to age matched non-impaired controls; in patients with normal WMS-III/MMSE, with subjective mild memory/mental status impairment, P300 latency was prolonged compared to controls (P=0.0004). The P300 latency increased by 0.72ms per year (P=7.9x10(-65)) and voltage decreased by 0.03dV per year (P=6.7x10(-10)), and both parameters were linearly correlated with the age of the subjects. Male subjects had an average voltage of 6.1dV and female 6.8dV (P=0.00009). Statistically, prolonged latency began at age range 41-50 (P=0.0002); reduced P300 voltage began at age range 51 60 (P=0.003). WMS-III memory decline for all measures began in females at age range 61-70 (P value at least=0.02) and for males at age range 61-80 (P=0.02). Prolonged P300 latency (P<0.0001) and memory impairment (at least <0.02) were greater for females than males. MMSE memory decline, male and female, began at age range 81-90 (P value of at least 0.00007). In our logistic regression model P300 latency was more predictive of WMS-III impairment than MMSE > 24. In patients whose WMS-III score is impaired < or = 69, or borderline < or = 79 (P at least=0.004), a P300 latency more prolonged than the norm (> or = 300 + 30 + Age) identifies these patients, whereas a MMSE > 24 failed. With the ROC curve, we confirmed that P300 latency could accurately identify borderline/impaired memory. PMID- 14521275 TI - Value of the early electroencephalogram after a first unprovoked seizure. AB - Studies on the predictive value of the electroencephalogram (EEG) concerning the risk of seizure recurrence have shown contradictory results. We prospectively studied the predictive value of the standard EEG and EEG with sleep deprivation for seizure relapse in adult patients presenting with a first unprovoked seizure. EEGs were performed on 157 adult patients within the first 48 hours of the first seizure. Additional EEGs with sleep deprivation were obtained in 60 cases. The standard EEG was abnormal in 70.7% and significantly associated with an increased risk of seizure recurrence [risk ratio 4.5, 95% confidence interval (CI) 1.8; 11.3, p=0.001]. Subgroup analysis revealed the highest recurrence rates for patients with focal epileptiform activity (risk ratio 2.2, CI 1.2; 4.2, p=0.01). EEGs with sleep deprivation were abnormal in 48.3% of all cases and revealed epileptiform discharges in 13.3% of the patients who had no epileptiform activity in the standard EEG. Routine EEG revealed abnormalities in 60 of 94 patients who presented with normal neurologic status on admission. Further neuroradiological examinations detected previously unknown brain lesions in 19 of these cases, particularly cerebrovascular disease (CVD, n=7), brain tumors (n=6), and posttraumatic scars (n=4). In conclusion, the EEG is important for the early detection of focal nonepileptic and epileptic abnormalities after a first unprovoked seizure in adult patients and may provide valuable information on previously unknown disorders, particularly CVD and cerebral tumors. The abnormal EEG is a highly significant predictor for seizure recurrence. An additional EEG with sleep deprivation is helpful in cases when standard EEG does not reveal epileptiform discharges. PMID- 14521276 TI - EEG and behavioral changes following neurofeedback treatment in learning disabled children. AB - Neurofeedback (NFB) is an operant conditioning procedure, by which the subject learns to control his/her EEG activity. On one hand, Learning Disabled (LD) children have higher values of theta EEG absolute and relative power than normal children, and on the other hand, it has been shown that minimum alpha absolute power is necessary for adequate performance. Ten LD children were selected with higher than normal ratios of theta to alpha absolute power (theta/alpha). The Test Of Variables of Attention (TOVA) was applied. Children were divided into two groups in order to maintain similar IQ values, TOVA values, socioeconomical status, and gender for each group. In the experimental group, NFB was applied in the region with highest ratio, triggering a sound each time the ratio fell below a threshold value. Noncontingent reinforcement was given to the other group. Twenty half-hour sessions were applied, at a rate of 2 per week. At the end of the 20 sessions, TOVA, WISC and EEG were obtained. There was significant improvement in WISC performance in the experimental group that was not observed in the control group. EEG absolute power decreased in delta, theta, alpha and beta bands in the experimental group. Control children only showed a decrease in relative power in the delta band. All changes observed in the experimental group and not observed in the control group indicate better cognitive performance and the presence of greater EEG maturation in the experimental group, which suggests that changes were due not only to development but also to NFB treatment. PMID- 14521277 TI - Patterns of interictal spike propagation across the central sulcus in benign rolandic epilepsy. AB - It has been reported that the rolandic area generating spikes is hyperexcitable, and that rolandic spikes propagate across the central area. However, the pattern of rolandic spike propagation and how the dipolar distribution of the spikes is related to the propagation pattern have not yet been studied. Thirty-nine EEGs from 27 patients with benign rolandic epilepsy (BRE) were examined. Sequential topographic mapping in 4-ms steps was used to analyze the pattern of spike propagation. The locations of maximum negative foci, the presence and distribution of the dipolar field, and the propagation pattern were examined. Dipoles were present in 23 (85.2%) out of 27 patients and in 43 (72.9%) out of 59 foci. Thirty-two foci (54.2%) in 20 patients demonstrated a propagation pattern. The typical pattern consisted of propagation from central to mid-temporal locations across the central sulcus. Most spike foci exhibiting a propagation pattern had a dipolar distribution (87.5%; p=0.008). These results suggest that rolandic spikes originate from sulcal or gyral cortices on either side of the central sulcus, and that spike propagation can ensue by intracortical spreading across the central sulcus. PMID- 14521278 TI - Tracing the pathways of neotropical migratory shorebirds using stable isotopes: a pilot study. AB - We evaluated the potential use of stable isotopes to establish linkages between the wintering grounds and the breeding grounds of the Pectoral Sandpiper (Calidris melanotos), the White-rumped Sandpiper (Calidris fuscicollis), the Baird's Sandpiper (Calidris bairdii), and other Neotropical migratory shorebird species (e.g., Tringa spp.). These species molt their flight feathers on the wintering grounds and hence their flight feathers carry chemical signatures that are characteristic of their winter habitat. The objective of our pilot study was to assess the feasibility of identifying the winter origin of individual birds by: (1) collecting shorebird flight feathers from several widely separated Argentine sites and analyzing these for a suite of stable isotopes; and 2) analyzing the deuterium and 18O isotope data that were available from precipitation measurement stations in Argentina. Isotopic ratios (delta13C, delta15N and delta34S) of flight feathers were significantly different among three widely separated sites in Argentina during January 2001. In terms of relative importance in separating the sites, delta34S was most important, followed by delta15N, and then delta13C. In the complete discriminant analysis, the classification function correctly predicted group membership in 85% of the cases (jackknifed classification matrix). In a stepwise analysis delta13C was dropped from the solution, and site membership was correctly predicted in 92% of cases (jackknifed matrix). Analysis of precipitation data showed that both deltaD and delta18O were significantly related to both latitude and longitude on a countrywide scale (p < 0.001). Other variables, month, altitude, explained little additional variation in these isotope ratios. Several issues were identified that will likely constrain the degree of accuracy one can expect in predicting the geographic origin of birds from Argentina. There was unexplained variation in isotope ratios within and among the different wing feathers from individual birds. Such variation may indicate that birds are not faithful to a local site during their winter stay in Argentina. There was significant interannual variation in the deltaD and delta18O of precipitation. Hence, specific locations may not have a constant signature for some isotopes. Moreover, the fractionation that occurs in wetlands due to evaporation significantly skews local deltaD and delta18O values, which may undermine the strong large-scale gradients seen in the precipitation data. We are continuing the research with universities in Argentina with a focus on expanding the breadth of feather collection and attempting to resolve the identified issues. PMID- 14521279 TI - Changes in hydrogen isotope ratios in sequential plumage stages: an implication for the creation of isotope-base maps for tracking migratory birds. AB - Accurate reference maps are important in the use of stable-isotopes to track the movements of migratory birds. Reference maps created by the analysis of samples collected from young at the nest site are more accurate than simply referring to naturally occurring patterns of hydrogen isotope ratios created by precipitation cycles. Ratios of hydrogen isotopes in the nutrients incorporated early in the development of young birds can be derived from endogenous, maternal sources. Base maps should be created with the analysis of tissue samples from hatchlings after local the isotopic signature of exogenous nutrients is dominant. Migratory species such as Peregrine Falcons are known to use endogenous sources in the creation of their eggs, therefore knowledge of what plumage stage best represents the local hydrogen ratios would assist in the planning of nest visits. We conducted diet manipulation experiments involving Japanese Quail and Peregrine Falcons to determine the plumage stage when hydrogen isotope ratios were indicative of a switch in their food source. The natal down of both the quail and falcons reflected the diet of breeding adult females. The hydrogen isotope ratios of a new food source were dominant in the juvenile down of the young falcons, although a further shift was detected in the final juvenile plumage. The juvenile plumage is grown during weeks 3-4 after hatch on Peregrine Falcons. Nest visits for the purpose of collecting feathers for isotope-base-map creation should be made around 4 weeks after the presumed hatch of the young falcons. PMID- 14521280 TI - Using stable isotopes in mangrove fisheries research--a review and outlook. AB - Mangrove forests are important coastal wetlands in most of the tropics, but their importance to fisheries is poorly understood. Rationales for conserving these ecosystems are partly based on ideas that mangroves are important to food webs that support fisheries. Here we review use of stable isotopes to test the strength of the mangrove forest-fish connection, suggesting that a dual focus on both food web ecology and habitat use may be most profitable in future studies of this type. PMID- 14521281 TI - Contribution of collagen stable isotope biogeochemistry to the paleobiology of extinct endemic vertebrates from Tenerife (Canary Islands, Spain). AB - The paleodiet and paleoenvironmental context of two extinct species from Tenerife island, one giant rat Canariomys bravoi and one giant lizard Gallotia goliath, have been investigated using carbon and nitrogen isotopic compositions of fossil bone collagen. Preliminary to this study, a calibration of the isotopic variations of bone collagen from modern Rat Rattus rattus, Rabbit Oryctolagus cuniculus and Lizard Gallotia galotti relative to environmental conditions on Tenerife Islands has been attempted. No clear relationship could be found between collagen delta13C and delta15N values and aridity; the only relevant factors seem to be seashore proximity for rat, and the relative amount of C3 and CAM plants. It seems that anthropic activities have interfered with the expected relationships between collagen isotopic compositions and environmental conditions. Most fossil specimens yielded well preserved collagen. The isotopic composition of giant rat and giant lizard collagen suggest a purely C3 environment, possibly more humid than today on Tenerife. Large ranges of nitrogen isotopic compositions, especially within giant rats, may be due to local environmental conditions. Further work is needed in order to provide more valuable paleobiological information in order to better understand the role of environmental factors in the evolution and extinction of insular endemic species on Tenerife. PMID- 14521282 TI - Comparative equilibration and online technique for determination of non exchangeable hydrogen of keratins for use in animal migration studies. AB - Stable hydrogen-isotope ratios (deltaD) of keratin provide a novel means for tracking geographical movements of birds and other species. Here we describe a rapid, low cost, analytical approach to facilitate online continuous-flow isotope ratio mass spectrometry (CF-IRMS) deltaD analyses of keratins (120-160 samples per day) through the use of calibrated keratin working standards and "comparative equilibration" to correct for the effects of moisture on exchangeable hydrogen. It is anticipated that this analytical approach and CF-IRMS will greatly aid in providing cost effective and directly comparable deltaD results on keratins and feathers among various laboratories and researchers involved in animal migration studies. PMID- 14521283 TI - Steady state models of stable isotopic distributions. AB - This study reviews common calculations and mathematical models used in stable isotopic studies. Some approximations are adopted to simplify the algebra for use in steady state models, with more rigorous mathematics outlined in an Appendix. Review shows that steady-state isotopic models are easy to construct, provide good approximations of system behavior, and are very helpful in evaluating isotopic cycling in many kinds of systems. PMID- 14521284 TI - Sample preparation techniques for the determination of natural 15N/14N variations in amino acids by gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). AB - In order to identify natural nitrogen isotope variations of biologically important amino acids four derivatization reactions (t-butylmethylsilylation, esterification with subsequent trifluoroacetylation, acetylation and pivaloylation) were tested with standard mixtures of 17 proteinogenic amino acids and plant (moss) samples using GC-C-IRMS. The possible fractionation of the nitrogen isotopes, caused for instance by the formation of multiple reaction products, was investigated. For biological samples, the esterification of the amino acids with subsequent trifluoroacetylation is recommended for nitrogen isotope ratio analysis. A sample preparation technique is described for the isotope ratio mass spectrometric analysis of amino acids from the non-protein (NPN) fraction of terrestrial moss. 14N/15N ratios from moss (Scleropodium spec.) samples from different anthropogenically polluted areas were studied with respect to ecotoxicologal bioindication. PMID- 14521285 TI - Good performance of a titanium femoral component in cementless hip arthroplasty in younger patients: 97 arthroplasties followed for 5-11 years. AB - We performed 97 uncemented primary total hip arthroplasties in 80 patients having an average age of 50 years. The femoral implant was a titanium stem with a proximal circumferential plasma spray-coating. Three different acetabular components were used: a threaded and partly porous-coated design in 70% of the cases. The average follow-up period was 8 years. 1 stem was revised 9 years after insertion due to a comminuted fracture of the proximal femur, 1 stem was revised 9 years after insertion due to a deep infection. No stem revisions were due to aseptic loosening. 1 femora had areas of distal osteolysis associated with a deep infection, but no signs of proximal loosening. 3 femora had areas of minor proximal osteolysis. 16 acetabular components (14 threaded) had been revised in 13 patients. The average Harris hip score was 91 points at the latest follow-up. We conclude that the uncemented titanium femoral component with a circumferential porous coating performed well in these patients, most of whom were young. As reported previously, aseptic loosening of threaded acetabular components was common. PMID- 14521286 TI - Alternative materials to improve total hip replacement tribology. AB - An improvement in tribology of bearing surfaces is an effective means of increasing the longevity of total hip replacement (THR). Currently, 3 approaches are available to achieve this aim: first, use of highly cross-linked UHMWPE; second, aluminum oxide ceramic bearings, and third, metal-on-metal bearings. Cross-linking reduces the wear resistance of UHMWPE markedly without impairment of other significant properties of the material. Simulator studies and some clinical long-term (10-22 years) follow-up surveys suggest an almost immeasurable wear of the highly cross-linked UHMWPE-based acetabular components during an expected clinical life span. Bioinert alumina ceramic (aluminum oxide) was introduced 3 decades ago for THR-bearing surfaces to improve performance and longevity. Alumina ceramic is entirely biostable and bioinert and has good mechanical properties. For correctly positioned alumina-on-alumina bearings, the annual linear wear rate has been reported to be 3.9 microm. Alumina heads have been successfully used in combination with polyethylene sockets, but as regards wear, the best results have been obtained with alumina-on-alumina bearings. In ceramic THR bearings, precise manufacture and contact surface geometry, including optimal clearance, are most important. For the currently available products, the component fracture risk is almost nonexistent (less than 1 per 1000). Metal-on metal bearings were used in the early stage of THR surgery, although not all old designs were successful. More recent analyses of the early series have shown the advantages of metal-on-metal to be better and have led to a renaissance of this articulation. Initially, stainless steel was used because it was easy to manufacture and polish. Current metal-on-metal bearings are based on cobalt chromium-molybdenum alloys with varying carbon contents. Such bearings are self polishing. Linear wear rates remain at the level of a few microm a year. An improvement in technology has increased the life span of the above three THR bearing systems. Although the technical solutions differ considerably, they all seem to improve clearly the tribology and longevity of the THR. Each of these bearing concepts will probably permit the use of larger head sizes, to reduce the risk of impingement and luxations. PMID- 14521287 TI - Untreated congenital and posttraumatic high dislocation of the hip treated by replacement in adult age: 22 hips in 16 patients followed for 1-8 years. AB - Between 1993 and 2001, we replaced 22 hips in 16 patients with high dislocation of the hip. All but 1 case was due to congenital dislocation. In all cases the femur was osteotomized below the lesser trochanter and a straight uncemented stem inserted in the medullary canal. The acetabular prostheses were inserted at the site of the original acetabulum, usually after augmenting the acetabular rim by using the medial half of the resected proximal femur. The lateral part of the proximal femur with the intact attachment of the gluteus medius muscle was transposed and fixed to the femoral diaphysis thereby restoring direction of muscle pull. The HHS score increased from a median value of 42 points preoperatively to a median of 86 points after median 25 months of follow-up. Limp, which before surgery was severe in all cases, was less marked or had disappeared at follow-up. The legs had been lengthened median 2.5 (1.0-4.5) cm. No postoperative infections occurred. Palsy or loss of sensory function was not observed in any patient. Dislocation was the commonest complication. It occurred in 3 hips, 2 of which had to be revised to ensure stability. PMID- 14521288 TI - Howse II cemented titanium metal-backed acetabular cups: poor 10-year results in 107 hips. AB - We report a retrospective clinical and radiographic review of 107 consecutive primary total hip replacements, using the cemented Howse II prosthesis. The review concentrates on the long-term performance of the acetabular cup, which was designed with a cylindrical titanium metal backing and a polyethylene liner. In all the hips in the series, this cup was used with a modular titanium straight femoral stem and a 32-mm diameter titanium head. At a mean follow-up of 9.8 (7 11) years, 28 hips had already been revised and 10 others were recognized as clinical or radiographic failures. The indication for revision in 26 of the hips was aseptic loosening and the mean time to revision was 7.2 (3.7-10.9) years This metal-backed cup has only 42% survival at 10 years, which seems to be related to certain of its design features. These include an inadequate thickness of polyethylene sterilized by gamma irradiation in air, a lack of mechanical bonding between the liner and its metal shell, and the high wear rate from the use of a 32-mm titanium femoral head. PMID- 14521289 TI - Insertion of tantalum beads in RSA of the hip: variations in incidence of extra osseous beads with insertion site. AB - Radiostereometric analysis (RSA) is a technique that can be used to measure the in-vivo micro-motion of the components of hip arthroplasty. It requires the insertion of tantalum beads into the bone permitting analysis of the radiographs. Extra-osseous beads reduce the usable bead pattern and previous studies have reported their incidence in the proximal femur as between 11% and 13% of all inserted beads. We reviewed the relative incidence of extra-osseous beads by examining 1038 radiographs of 97 patients who were part of an ongoing RSA hip study. Extra-osseous beads were seen in 44% of our patients with most having a single extra-osseous bead. The relative incidence of extra-osseous beads was 2% and 6% in the femur and pelvis, respectively. We observed a reduction in the incidence of the femoral beads with time, but not of the pelvis, which suggests that experience in the insertion technique improves the insertion rates at least of the femur. Our findings indicate that the incidence of extra-osseous beads is lower than previously reported. However, care should be taken in the design of studies to ensure that an adequate number of beads are placed in locations which are both surgically achievable and technically useful. PMID- 14521290 TI - Open versus arthroscopic subacromial decompression: a prospective, randomized study of 34 patients followed for 8 years. AB - In a randomized prospective study, we selected 15 patients for arthroscopic subacromial decompression (ASD) and 19 patients for open subacromial decompression (OSD). All had impingement syndrome (Neer grade II), and had been unsuccessfully treated without surgery for more than 6 months. The UCLA Shoulder Rating Scale, Visual Analogue Scales for pain and satisfaction, isokinetic dynamometer recordings and physical testing were assessed preoperatively and at 1 (except isokinetic testing), 3, 6, and 12 months, and, finally, 8 years after surgery. We found essentially no differences in the clinical tests between the groups during this period. The use of ASD or OSD seems to be a matter of cosmesis and personal preference. PMID- 14521291 TI - Morphologic evaluation of the ulna. AB - Intramedullary ulnar nailing may be technically difficult. We used various methods and measurements to determine the ideal nail entry point and the shape, length, diameter and curvature of the medullary canal in 44 human cadaver ulnas. We found that the ideal nail entry point was, on average, 7 mm proximal and 3 mm lateral to the most prominent area of the olecranon. A nail of 3 mm diameter could easily be inserted through a hole at the proximal-lateral side of the most prominent part of the olecranon, but only 20% of all nails could be easily inserted through a hole in the middle of the olecranon. We found that correct selection of a nail of proper length and diameter, as well as an ideal nail entry point on the olecranon are essential to successful nailing. PMID- 14521292 TI - Radioulnar convergence after distal ulnar resection: mechanical performance of two commonly used soft tissue stabilizing procedures. AB - Resection of the distal ulna (Darrach operation) is a common method for salvaging the arthrotic distal radioulnar joint (DRUJ). However, problems have been reported with this procedure due to residual instability and radioulnar convergence. As a result, several methods of soft tissue stabilization for the unstable distal ulna have been developed. Although their clinical efficacy has been reported, biomechanical investigations of these procedures have not been reported. The purpose of our study was to evaluate the dynamic effects on radioulnar convergence and dorsal-palmar displacement of three procedures: the Darrach procedure, a pronator quadratus interposition flap and an extensor and flexor carpi ulnaris tenodesis. We tested 7 fresh-frozen cadaver upper extremities using a dynamic computer-controlled device that generated forearm rotation with physiologic loading of relevant muscles. Displacement data concerning the ulna relative to the radius through the range of forearm rotation was collected for 4 experimental conditions: intact, distal ulna resection alone, distal ulna resection with pronator quadratus interposition and distal ulna resection with extensor and flexor carpi ulnaris tenodesis. Distal ulna resection altered the kinematics, most predictably creating a convergence of the radius towards the ulna. Anteroposterior translations in each loading condition could be detected as well. The interposition of the pronator quadratus muscle or tenodesis with the extensor and flexor carpi ulnaris tendons did not reduce the radioulnar convergence created by resection of the distal ulna. PMID- 14521293 TI - Semimembranosus tendonitis after total knee arthroplasty: good outcome after surgery in 6 patients. AB - More than 700 total knee arthroplasties were performed in our Department between 1992-2000. 6 patients developed pain on the postero-medial aspect of the knee, within 1 year of surgery. The pain increased during walking and especially on rising from a chair. All patients were women with good knee motion and good alignment of the implanted prosthesis. We found no signs of low-grade infection. Injection of 1% Xylocaine relieved pain for a short time in all patients. They were all treated with excision of the reflected portion of the semimembranosus tendon which was not cut during the arthroplasty. Pain was completely relieved in 5 cases and improvement occurred in 1. PMID- 14521294 TI - Increased external tibial torsion in Osgood-Schlatter disease. AB - We studied the relationship between Osgood-Schlatter disease and torsional abnormalities of the lower limb in 21 boys with this condition and 20 age- and sex-matched controls. 3 groups of knees (20 control knees, 21 symptomatic and 21 asymptomatic or less symptomatic knees) were subjected to clinical, radiographic and CT evaluation. We found no statistically significant differences between patients and controls, as regards femoral anteversion, patellar congruence angle, patellar tilt angle and anterior tibial tuberosity-trochlear groove distance, but the condylomalleolar angle and tibial torsion angle were greater in patients. We found no differences between symptomatic and asymptomatic or less symptomatic knees in any of the parameters. All the symptomatic knees were on the side preferentially involved in jumping and sprinting. This increase in external tibial torsion may play a role as a predisposing mechanical factor in the onset of Osgood-Schlatter disease in male athletes. PMID- 14521295 TI - Meniscectomy leads to an early increase in subchondral bone plate thickness in the rabbit knee. AB - We evaluated morphological changes in the tibial bone after meniscectomy in a rabbit model. 15 rabbits subjected to a medial meniscectomy in the right knee and a sham-operation in the left. Histomorphometric parameters were evaluated in the subchondral bone plate and the underlying trabecular bone, 13, 25 and 40 weeks after surgery. 5 rabbits were used as unoperated controls. Meniscectomized knees had a thicker subchondral bone plate than sham-operated contralateral ones in 13 of the 15 rabbits (p = 0.01), but the trabecular bone showed no morphological differences. The meniscectomized knees of these rabbits developed mild osteoarthrosis, described elsewhere, which may have been partly due to a change in the mechanical properties of the thickened subchondral bone plate. Our findings suggest that the first bony response after meniscectomy occurs in the subchondral bone plate rather than in the trabecular bone. PMID- 14521296 TI - The effect of body mass and physical activity on the development of guinea pig osteoarthrosis. AB - We quantitatively evaluated the morphological and biochemical effects of body mass and physical activity on spontaneously developing guinea pig osteoarthrosis (OA). 6-month-old male guinea pigs were allocated to 3 groups: controls (C) living under standard laboratory conditions with food ad libitum; mobilized animals (M) allowed unrestricted motion in large rooms with food ad libitum; and a diet group (D) weight-matched with the M-group. At 9- and 12-months of age they were killed and the left proximal tibia was processed for quantitative histology and the right tibial articular cartilage for analyses of glycosaminoglycan (GAG). OA mostly occurred on the medial condyle's central part not covered by the meniscus. The thinnest cartilage was found in 12-month-old M-animals (M12), which had 60% of the central cartilage surface affected by lesions that extended down to the mineralized cartilage. C12 had 25% exposed mineralized cartilage and D12, 2%. Subchondral bone density followed the loading patterns--the highest in M12 and lowest in D12. M12 had the lowest cartilage GAG concentrations. Load appears to be a key external factor in guinea pig OA. An increase in physical activity may be chondroprotective in the early phase, but harmful when fibrillations eventually have developed. This is underscored by the extensive OA changes in M12, although these animals weighed about the same as D12 (which had the least extensive OA). Therefore, a reduction in body mass seems to retard the progression of OA in animals, which are mainly subjected to a static load (C12 and D12), but not sufficiently in animals with a more dynamic load (M12). Changes in morphological patterns are paralleled by changes in GAG concentration, which probably reflect the metabolic capacity of the cartilage. PMID- 14521297 TI - Nonmetastatic osteosarcoma of the extremity with pathologic fracture at presentation: local and systemic control by amputation or limb salvage after preoperative chemotherapy. AB - To determine whether a pathologic fracture in osteosarcoma of long bones has prognostic importance, and limb salvage can be safely performed in such cases, we reviewed the surgical treatment and oncologic results in 46 patients with nonmetastatic osteosarcoma of the extremity and pathologic fracture at presentation who had been treated in our Institution with neoadjuvant chemotherapy, between 1983 and 1999. Neoadjuvant chemotherapy was given according to 6 consecutive protocols. Surgery consisted of limb salvage (34 patients), amputation (11 patients) and rotationplasty (1 patient). The average follow-up was 11 (3-20) years. 28 patients remained continuously disease-free, 17 patients relapsed and 1 died of chemotherapy-related toxicity. Despite the high rate of limb salvage, only 2 local failures occurred, 1 after amputation and 1 after limb salvage. The 5-year disease-free survival and overall survival rates were 59% and 65%, respectively, with no differences between amputated and resected patients. These results are similar to those obtained in 689 contemporary patients having an osteosarcoma without a pathologic fracture treated in our Institution, and using the same protocols for chemotherapy. We conclude that with neoadjuvant chemotherapy, osteosarcoma patients presenting with a pathologic fracture can be surgically treated like those with no fracture, and that limb salvage procedures do not increase the risk of local recurrence or death of these patients. PMID- 14521298 TI - Desmoid tumors: a clinical review of 30 patients with more than 20 years' follow up. AB - We retrospectively studied the outcome in a consecutive series of 30 patients with desmoid tumors who were followed for more than 20 years after treatment (surgery in 29 patients). A local recurrence occurred in 12 patients and more than 1 recurrence occurred in 8. 3 patients had spontaneous complete regression of the tumor. At follow-up after mean 28 (20-54) years, all patients were tumor free except 1 who had had a stable tumor for at least 11 years. Symptoms were related more to treatment than to the tumor. We conclude that desmoid tumors may have a high capacity for self-limitation and that conservative therapy should be considered in symptom-free patients. PMID- 14521299 TI - Bone marrow perfusion in healthy subjects assessed by scintigraphy after application of a tourniquet. AB - By means of a gamma camera technique involving a bolus injection of autologous 99mTc-labeled erythrocytes, we found that despite a tourniquet on the limb, the erythrocytes entered the bone marrow, and caused oozing in both lower (10 subjects) and upper limbs (14 subjects). The 24 healthy volunteers, 18 men, had a mean age of 50 (range 20-86) years. The activity reached the distal femur in a median of 365 (quartile 278-560) seconds and the distal humerus in a median of 280 (quartile 208-370) seconds. The median velocity for movement of erythrocytes in the femur was 3.7 (quartile 2.74.4) cm/min and in the humerus, it was 4.0 (quartile 3.1-5.3) cm/min. In 21 subjects, this activity reached the periarticular soft tissue. After 15 min with the tourniquet inflated, the activity in the distal femur of the tourniquet limb was 9 (quartile 5-18)% of that in the limb without a tourniquet. This study shows that although a tourniquet effectively occludes the extra-osseous blood supply, some intra osseous blood supply is retained, which makes it difficult to obtain a bloodless field in some patients, despite the use of a tourniquet. PMID- 14521301 TI - Good preservation of initial mechanical properties in lipid-extracted, disinfected, freeze-dried sheep patellar tendon grafts. AB - Patellar tendon allografts can be used for anterior cruciate reconstruction to avoid morbidity of autografts on the donor side. Secondary processing of allografts is important for reducing immunological reactions, bacterial contamination and improving storage. We analyzed the effects of processing on the mechanical properties of patellar tendon grafts in 20 sheep. Group I (n = 10) was deep-frozen at -80 degrees C. Group II (n = 10) was processed by a lipid extraction/ freeze-drying method, including iodoacetic acid disinfection. The contralateral tendons, freeze-dried by dehydration in a vacuum at -50 degrees C for 3 hours, served as controls. We measured failure stress: group I (53, SD 14 MPa), control (26, SD 15 MPa) (p = 0.04); group II (49, SD 13 MPa), control (28, SD 5 MPa) (p = 0.08). Failure strain, normalized stiffness, and energy to failure were similar in the groups. Our method of extended processing did not change the properties of the deep-frozen patellar tendons. Therefore in vivo experiments can be used when studying the effects of transplantation on mechanical properties. PMID- 14521300 TI - Indomethacin and celecoxib improve tendon healing in rats. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit the formation of bone. However, they have been shown to increase tensile strength in healing tendons. Most NSAIDs inhibit two isoforms of cyclooxygenases called Cox-1 and Cox-2. Thanks to fewer side-effects, the recently introduced selective cyclooxygenase-2 (Cox-2) inhibitors will probably promote more widespread use of this kind of drug. To clarify the effects on tendon healing of a general Cox-inhibitor (indomethacin) as well as a selective Cox-2 inhibitor (celecoxib), we resected 3 mm of the Achilles tendon in rats and measured the strength of the tendon regenerate. Indomethacin given as daily injections in doses of 1.5, 3.0 and 5.0 mg/kg reduced the thickness (cross-sectional area) of the tendon regenerate at 14 days, as compared to controls, but there was no difference in the failure load or stiffness. In another series of measurements, indomethacin in a dose of 3.0 mg/kg reduced the cross-sectional area at 10, 14 and 18 days after transsection. Failure load was not affected, but tensile stress at failure was increased by indomethacin at 14 and 18 days. Indomethacin (3 mg/kg) was then compared to celecoxib (4.5 mg/kg) and controls 14 days after tendon transsection. No difference between the drugs was seen. Again, the transverse area was smaller in the treated tendons than in the controls. Failure load was unchanged and the tensile stress was higher in the treated tendons than in the controls. Because of the reduction in cross-sectional area without an effect on failure load, the use of Cox-inhibitors may be beneficial in clinical situations where thickening of a healing tendon is a problem--e.g., in the hand or shoulder. PMID- 14521302 TI - An external fixation method and device to study fracture healing in rats. AB - We wished to establish a reproducible model for fracture fixation to be used in fracture healing research and therefore developed an external fixation construct and surgical procedure adapted to Sprague-Dawley rats. We evaluated the mechanical properties of the construct in brass rods and rat bone, in an Instron test machine with axial and transverse loading, and the in vivo performance. We found that the mechanical properties of the construct in brass rods were predictable and could be repeated in rat femora. In all tests, the axial load was about 10 times the transverse for the same degree of deformation. The stiffness among fixators was uniform. 1 mm pins caused about 50% less stiffness than 1.2 mm pins in axial loading of rat bone (p < 0.001) and brass rods (p < 0.001) as well as in transverse loading of brass rods (p < 0.001). Loosening of 1 or 2 screws that lock the pins to the fixator reduced stiffness by about 50% in axial loading of rat bone (p = 0.009) and brass rods (p = 0.05). A change in the distance between the bone surface and the fixator was linearly related to the stiffness in axial loading of rat bone (p < 0.001) and brass rods (p < 0.001) and in transverse loading of brass rods (p < 0.001). If the bone ends touched each other, the axial stiffness of the construct increased almost 10 times (265 N/mm), as compared to a fracture gap size of 2 mm (31 N/mm). In vivo experiments had a complication rate of less than 10% when we used 1.2 mm pins, 6 mm offset and rats weighing 350-450 g. Our method and device for experimental external fixation of rat femora are reliable and the findings are reproducible. These can be used in bone repair and fracture healing research. PMID- 14521303 TI - A combination of bisphosphonate and BMP additives in impacted bone allografts. AB - OP-1 increases bone ingrowth distance of new bone into allografts (Tagil et al. 2000), but the bone density after incorporation may be reduced by an increase in resorption (Hoistner et al. 2000). Bisphosphonates inactivate osteoclasts and can be used to increase allograft bone density after incorporation (Aspenberg and Astrand 2002). A combination of locally-applied bisphosphonate and OP-1 in the graft could therefore be expected to increase both new bone ingrowth and density. We tested this by using a rat bone chamber model. OP-1 alone increased the ingrowth distance of bone. Clodronate increased final bone density greatly, but reduced the ingrowth distance of new bone into grafts that were extremely impacted. This reduction was improved by adding OP-1. Regardless of graft density, combinations of OP-1 and clodronate included a high final bone density, but the ingrowth distances were shorter than with OP-1 alone. These data indicate that new bone and tissue ingrowth into a compacted graft depends on resorption and that resorption is a prerequisite for the stimulating effect of OP-1 in this experimental set-up. Although the problems associated with the use of OP-1 in impaction grafting may be solved by adding a bisphosphonate, some of the benefits of OP-1 can be lost. PMID- 14521304 TI - Optimal handling of fresh cancellous bone graft: different peroperative storing techniques evaluated by in vitro osteoblast-like cell metabolism. AB - We investigated the influence of three peroperative handling techniques on the quality of autogenous bone graft by means of osteoblast-like cell metabolism in vitro. Cancellous bone was harvested from the iliac crest of 12 4-month-old female pigs. Osteoblast-like cell cultures were established, using the tissue explant method: (1) immediately after harvest of bone, (2) after storage of bone in saline at room temperature for 2 hours and (3) after "dry" storage of bone at room temperature for 2 hours. Proliferation was assessed by 3H-thymidine incorporation. Differentiation was assessed by alkaline phosphatase activity and procollagen I production (PICP). We found that osteoblast-like-cell proliferation was higher, when cultures were started shortly after harvesting of bone, or else stored in saline for 2 hours, as compared to bone left "to dry" for 2 hours. Basal alkaline phosphatase and PICP production did not differ in the three groups. These in vitro results suggest the superiority of harvest of autogenous bone graft shortly before the grafting procedure, or else temporary storage of the graft in saline for up to 2 hours. PMID- 14521305 TI - Disorders associated with osteopoikilosis: 5 different lesions in a family. AB - Although osteopoikilosis is generally considered an accidental finding, several developmental dysplasias coexisting with this disorder have been reported. However, all authors have mentioned only one coexisting finding, and most of them are case reports. We report a family in whom various members had osteopoikilosis with 5 different associated lesions. We suggest that osteopoikilosis is a bone manifestation of a generalized fibroproliferative or stenosing disease. PMID- 14521306 TI - Late symptoms after hip fracture with displacement of the lesser trochanter--a case report. PMID- 14521307 TI - Determination of chlorophenols in soil samples by microwave-assisted extraction coupled to headspace solid-phase microextraction and gas chromatography-electron capture detection. AB - Microwave-assisted extraction coupled to headspace solid-phase microextraction was studied and applied for one-step in-situ sample preparation prior to analysis of chlorophenols (CPs) in soil samples. The CPs in soil sample were extracted into the aqueous solution and then directly onto the solid-phase microextraction (SPME) fiber in headspace under the aid of microwave irradiation. After being desorbed from SPME fiber in the GC injection port, CPs were analyzed with a GC electron-capture detection system. Parameters affecting the extraction efficiency such as the extraction solutions, the pH in the slurry, the humic acid content in the soil, the power and the irradiation time of microwave as well as the desorption parameters were investigated. Experimental results indicated that the extraction of a 1.0 g soil sample with a 6-ml aqueous solution (pH 2) and a polyacrylate fiber under the medium-power irradiation (132 W) for 9 min achieved the best extraction efficiency of about 90% recovery and less than 10% RSD. Desorption was optimal at 300 degrees C for 3 min. Detection limits were obtained at around 0.1-2.0 microg/kg levels. The proposed method provided a simple, fast, and organic solvent-free procedure to analyze CPs from soil sample matrix. PMID- 14521308 TI - Ultrasound-assisted extraction of soy isoflavones. AB - Efficiency in extracting four isoflavone derivatives (daidzin, glycitin, genistin and malonyl genistin) from freeze-dried ground soybeans was compared for mix stirring extraction and ultrasound-assisted extraction, using different solvents and extraction temperatures with both. The efficiency of the extraction of soy isoflavones was improved by ultrasound but was dependent on the solvent employed. Optimization of the ratios of sample quantity to solvent volume and length of extraction time was also performed. Isoflavones can be quantitatively extracted from soybeans with 50% ethanol at 60 degrees C using ultrasound-assisted extraction in 20 min. PMID- 14521309 TI - Determination of organochlorine pesticides in water using microwave assisted headspace solid-phase microextraction and gas chromatography. AB - A coupled technique, microwave-assisted headspace solid-phase microextraction (MA HS-SPME), was investigated for one-step in situ sample pretreatment for organochlorine pesticides (OCPs) prior to gas chromatographic determination. The OCPs, aldrin, o,p'-DDE, p,p'-DDE, o,p'-DDT, p,p'-DDT, dieldrin, alpha-endosulfan, beta-endosulfan, endosulfan sulfate, endrin, delta-HCH, gamma-HCH, heptachlor, heptachlor epoxide, methoxychlor and trifluralin were collected by the proposed method and analyzed by gas chromatography with electron-capture detection (GC ECD). To perform the MA-HS-SPME, six types of SPME fibers were examined and compared. The parameters affecting the efficiency in MA-HS-SPME process such as sampling time and temperature, microwave irradiation power, desorption temperature and time were studied to obtain the optimal conditions. The method was developed using spiked water samples such as field water and with 0.05% humic acid in a concentration range of 0.05-2.5 microg/l except endosulfan sulfate in 0.25-2.5 microg/l. The detection was linear over the studied concentration range with r2>0.9978. The detection limits varied from 0.002 to 0.070 microg/l based on S/N=3 and the relative standard deviations for repeatability were <15%. A certified reference sample of OCPs in aqueous solution was analyzed by the proposed method and compared with the conventional liquid-liquid extraction procedure. These results are in good agreement. The results indicate that the proposed method provides a very simple, fast, and solvent-free procedure to achieve sample pretreatment prior to the trace-level screening determination of organochloride pesticides by gas chromatography. PMID- 14521310 TI - Comparison between adsorption isotherm determination techniques and overloaded band profiles on four batches of monolithic columns. AB - The adsorption isotherms of 4-tert.-butyl phenol were measured on four different monolithic columns, using three different techniques, classical frontal analysis (FA), the perturbation on a plateau method (PP) and the recently introduced numerical procedure known as the inverse numerical method (IN). This last approach requires only the recording of a few overloaded profiles and has the potential advantage of affording a dramatic decrease of the amounts of compounds, solvent, and time needed to determine accurate estimates of the coefficients of the isotherm. The reproducibility of the adsorption data measured on the four columns is discussed with reference to the specific techniques used for obtaining these data and to the most suitable equation used for modeling them. The data obtained for the different columns were highly consistent. The inverse numerical approach was confirmed to provide a powerful, accurate, and economic method for measuring single component adsorption data. PMID- 14521311 TI - High-performance liquid chromatographic analysis of secoisolariciresinol diglucoside and hydroxycinnamic acid glucosides in flaxseed by alkaline extraction. AB - A HPLC method was developed for the analysis of secoisolariciresinol diglucoside (SDG) and hydroxycinnamic acid glucosides in milled defatted flaxseed flour. Direct extraction by 1 M NaOH for 1 h at 20 degrees C resulted in a higher yield than that obtained by hydrolysis of alcoholic extracts. An internal standard, o coumaric acid, was used and the method was found to be easy, fast, and with good repeatability. On dry matter basis, different samples of flaxseeds varied considerably in their content of (+)-SDG (11.9-25.9 mg/g), (-)-SDG (2.2-5.0 mg/g), p-coumaric acid glucoside (1.2-8.5 mg/g), and ferulic acid glucoside (1.6 5.0 mg/g). PMID- 14521312 TI - Determination of tricarboxylic acid cycle acids and other related substances in cultured mammalian cells by gradient ion-exchange chromatography with suppressed conductivity detection. AB - An ion-exchange chromatography method was established for simultaneously analyzing the tricarboxylic acid (TCA) cycle acids and other related substances in cultured mammalian cells, including citrate, cis-aconitate, isocitrate, alpha ketoglutarate, succinate, malate, fumarate, oxaloacetate, trans-aconitate, phosphate, lactate and pyruvate. A Dionex 600 ion chromatograph with an ion suppressor and a conductivity detector, and an IonPac AS11-HC analytical column were employed. An NaOH gradient elution containing 13.5% methanol contributed to sufficient separation of target substances. The stability of carboxylic acids was investigated and oxaloacetate was found to be extremely unstable especially at pH 3. TCA cycle acids and other related substances in Chinese hamster ovary (CHO) cells were separated completely, and lactate, malate, phosphate, citrate and cis aconitate were quantified due to their higher concentrations. In the quantification of the five substances, detection limits (S/N=3) ranged from 0.12 to 0.48 microM, the correlation coefficients from 0.9982 to 1.0000 in their linear ranges of concentration, and the recoveries from 87 to 95%. The metabolic status of CHO cells was analyzed on the basis of the intracellular concentrations of TCA cycle acids. PMID- 14521313 TI - Simultaneous determination of 5-hydroxyindoles and catechols by high-performance liquid chromatography with fluorescence detection following derivatization with benzylamine and 1,2-diphenylethylenediamine. AB - A highly selective and sensitive method for the simultaneous determination of 5 hydroxyindoles and catechols (serotonin, norepinephrine, dopamine and related compounds) by high-performance liquid chromatography with fluorescence detection is described. The method is based on the two-step precolumn derivatization of 5 hydroxyindoles and catechols with benzylamine (BA) and 1,2 diphenylethylenediamine (DPE), respectively, resulting in highly fluorescent and stable benzoxazole derivatives. The first derivatization with BA proceeds at room temperature (ca. 23 degrees C) for 2 min in a mixture of 0.3 M 3-cyclohexylamino 1-propanesulfonic acid buffer (pH 10.0) and methanol in the presence of potassium hexacyanoferrate(III). The subsequent second derivatization with DPE is carried out at 50 degrees C for 20 min in the presence of glycine. The resulting fluorescent derivatives of five 5-hydroxyindoles and seven catechols are separated on a reversed-phase column (150 x 1.5 mm I.D., packed with C18 silica, 5 microm) with isocratic elution using a mixture of acetonitrile-15 mM acetate buffer (pH 4.5) (34:66, v/v) containing 1 mM octanesulfonic acid sodium salt, and are detected spectrofluorimetrically at 480 nm with excitation at 345 nm. The detection limits (signal-to-noise ratio of 3) of the related compounds are 80 amol to 86 fmol for a 20-microl injection. PMID- 14521314 TI - Investigation of natural dyes occurring in historical Coptic textiles by high performance liquid chromatography with UV-Vis and mass spectrometric detection. AB - Liquid chromatography (LC) combined with ultraviolet-visible (UV-Vis) and mass spectrometric (MS) detection was utilized to study the chemical components present in extracts of natural dyes originating from fiber samples obtained from Coptic textiles from Early Christian Art Collection of National Museum in Warsaw. Chromatographic retention, ionization, UV-Vis and mass spectra of twenty selected dye compounds of flavanoid-, anthraquinone- and indigo-types were studied. Most of the investigated compounds could be ionized by positive and negative ion electrospray ionization. Difficulties with the ionization by electrospray were experienced for indigotin and brominated indigotins, but these were ionized by atmospheric pressure chemical ionization. Mass spectrometric detection, utilizing different scanning modes of a triple quadrupole mass spectrometer, combined with the UV-Vis detection was demonstrated to be a powerful approach to detection and identification of dyes in the extracts of archeological textiles. Using this approach the following compounds were identified in the extracts of Coptic textiles: luteolin, apigenin, rhamnetin, kaempferol, alizarin, purpurin, xanthopurpurin, monochloroalizarin, indirubin, and so the type of dye that was utilized to dye the textiles could be identified. Detection capabilities for several dye-type analytes were compared for the UV-Vis and mass spectrometric detection. The signal-to-noise ratios obtained for luteolin, apigenin, and rhamnetin were higher for the MS detection for most of the examined sample extracts. Purpurin, alizarin, and indirubin showed similar signal-to-noise ratios for UV-Vis and mass spectrometric detection. PMID- 14521315 TI - Gas chromatographic-mass spectrometric study of the oil fractions produced by microwave-assisted pyrolysis of different sewage sludges. AB - The pyrolysis of sewage sludge was studied in a microwave oven using graphite as microwave absorber. The pyrolysis temperature ranged from 800 to 1000 degrees C depending on the type of sewage sludge. A conventional electrical furnace was also employed in order to compare the results obtained with both methods. The pyrolysis oils were trapped in a series of condensers and their characteristics such as elemental analysis and calorific value were determined and compared with those of the initial sludge. The oil composition was analyzed by GC-MS. The oils from the microwave oven had n-alkanes and 1-alkenes, aromatic compounds, ranging from benzene derivatives to polycyclic aromatic hydrocarbons (PAHs), nitrogenated compounds, long chain aliphatic carboxylic acids, ketones and esters and also monoterpenes and steroids. The oil from the electric oven was composed basically of PAHs such as naphthalene, acenapthylene, phenanthrene, fluoranthene, benzo[a]anthracene, benzofluoranthenes, benzopyrenes, indenepyrene, benzo[ghi]perylene, and anthanthrene. In contrast, these compounds were not produced in the case of microwave-assisted pyrolysis. PMID- 14521316 TI - Direct extraction of alkylphenols, chlorophenols and bisphenol A from acid digested sediment suspension for simultaneous gas chromatographic-mass spectrometric analysis. AB - The direct extraction of alkylphenols, chlorophenols and bisphenol A from an acid digested sediment suspension for GC-MS analysis was studied. The sediment was digested with acid while the hydrolyzed analytes were being extracted with dichloromethane. The conditions of the acid digestion and extraction were optimized in terms of time, acidity of digestion, and extracting solvent. It is possible to complete the extraction within 20 min with 5 ml of 0.1 M HCl digesting solution and three portions of 5 ml of dichloromethane. The recoveries of analytes were mostly around 90% with about 10% relative standard deviations. With this technique parallel treatment of large numbers of sediment samples is possible without any expensive special equipment or heating process. The analytical characteristics of this extraction technique were compared with Soxhlet extraction and the pressurized liquid extraction technique. The technique was examined and evaluated for real environmental sediment samples and certified reference material of natural matrix. PMID- 14521317 TI - Evaluation of immature granulocyte counts by the XE-IG master: upgraded software for the XE-2100 automated hematology analyzer. AB - We evaluated an automated immature granulocyte (IG) count in the peripheral blood with the XE-IG Master (Sysmex Corporation). The XE-IG Master is the upgraded software package for the XE-2100 automated hematology analyzer. Reproducibility tests demonstrated a mean coefficient of variation of 7.02% for the IG percentage (IG%) and 6.93% for the absolute IG count. Correlations of the IG counts were assessed in two ways. A flow cytometric IG count using CD11b, CD16, and CD45 monoclonal antibodies and a manual differential count were used as reference methods. The regression equation and the correlation coefficient of the IG% for the flow cytometric reference count versus results with the XE-IG Master were: y = 0.91x + 0.10; r = 0.96. For the comparison with the manual differential count of promyelocytes, myelocytes, and metamyelocytes, the regression equation and correlation coefficient were: y = 0.81x + 1.27; r = 0.78. Samples were found to be stable up to 48 hours both at room temperature and when refrigerated. We investigated the clinical significance of the IG count as a new marker of acute inflammation. In this preliminary study, most samples with a high IG count had positive values for C-reactive protein and the erythrocyte sedimentation rate (positive sample rates were 84.0% and 95.0%, respectively) despite neutrophil counts within the normal range. Elevated IG counts correlated most closely with CD64 expression on polymorphonuclear cells and less so with the concentration of interleukin 6. Compared with other available inflammation markers, the IG count is rapidly generated with each full blood count at no extra cost and with no delay in sample analysis. PMID- 14521318 TI - Heparin resistance. AB - Heparin therapy is gaining more use in many clinical conditions. Considering the serious nature of the diseases managed with heparin, it is necessary to achieve the therapeutic goal within a short time to gain the best possible benefit. Adequate anticoagulation also is necessary during procedures such as cardiopulmonary bypass and hemodialysis. Heparin resistance, inability to achieve therapeutic anticoagulation, has been described in as many as 22% of patients undergoing open-heart surgeries. The results of undercoagulation can vary from subclinical coagulation to death. It also is necessary not to over anticoagulate a patient and to understand the existence of alternative methods for monitoring heparin therapy to prevent excess heparin therapy and its complications. Many common dinical situations can induce resistance to the action of heparin. Being aware of the existence of heparin resistance and taking timely appropriate action would prevent catastrophic consequences. The use of antithrombin III is one way of preventing heparin resistance. PMID- 14521319 TI - Assessment of in vitro platelet activation by Advia 120 platelet parameters. AB - The Advia 120 Hematology System provides new platelet parameters that have been proposed as useful markers of platelet activation. The aim of this study was to investigate platelet parameter variations after adenosine diphosphate (ADP) activation of platelet-rich plasma (PRP) in vitro, with particular interest in the mean platelet component (MPC), which was compared with two well-known degranulation antigens. Changes in platelet parameters that were induced by the activation of PRP with different concentrations of ADP were examined first. The time course of parameter values up to 60 minutes after maximal ADP activation and the relationships between the MPC and P-selectin and granulophysin expression as determined by flow cytometry were then investigated. After 10 minutes of ADP stimulation, the MPC presented a dose-dependent increase. At the maximal ADP concentration, the initial increase of the MPC was followed by a progressive decrease, leading the MPC to become significantly lower with respect to the baseline after 60 minutes of incubation. Significant variations in other parameters are also described. Finally, a negative correlation was found between the MPC change with respect to time 0 and both P-selectin and granulophysin expression. The present study suggests that platelet parameter variation, particular in the MPC, may be used to assess platelet activation in PRPs stimulated by ADP. PMID- 14521320 TI - An abnormal activated partial thromboplastin time clotting waveform is associated with high mortality and a procoagulant state. AB - We studied the associations of a biphasic waveform identified in 61 consecutive patients by activated partial thromboplastin times performed with an MDA 180 coagulation analyzer (bioMerieux Inc., Durham, NC, USA). The patients were encountered in a wide range of locations, including medical and surgical services, emergency and outpatient departments, and intensive care unit. The patients encountered also had a variety of clinical diagnoses. Of the patients, 42.6% were dead on follow-up evaluation at 3 months, and 67.2% had a clinical condition known to predispose to the development of disseminated intravascular coagulation (DIC). Sepsis was the most frequent associated condition and was present in 54% of the patients. Only 6 patients had overt DIC according to a standardized scoring system. However, D-dimer was elevated in 90% of patients, and antithrombin III and protein C levels decreased in 75.4% and 63.9%, respectively. The platelet count was decreased in 26%. Our results indicated that a biphasic waveform is associated with high mortality in general hospital patients and is frequently associated with sepsis. This waveform appears to be associated with a consumptive procoagulant state, which may represent early or nonovert DIC. PMID- 14521321 TI - Performance evaluation of the Abbott Cell-Dyn 1800 automated hematology analyzer. AB - The Cell-Dyn 1800 is a new automated hematology analyzer that provides an 18 parameter blood count and operates at a throughput of 60 specimens per hour. (Clinical significance has not been established for plateletcrit and platelet distribution width; therefore these parameters are not reportable in the United States.) The instrument is smaller than its predecessor and offers a variety of enhanced features, including increased patient data storage, availability of a bar code reader, and a cyanide-free reagent system. Our study comprehensively assessed the analytical characteristics of the instrument in a variety of areas. When applicable the requirements of appropriate NCCLS guideline documents were followed. Instrument background and carryover were shown to be consistent with the performance specification. Accuracy was demonstrated by linearity studies and by comparison of results from the automated blood count parameters of the Cell Dyn 1700 and from the manual differential method. The instrument-generated results and morphology flags were found to be effective in screening for abnormalities demonstrated by manual morphological assessment. Precision of the Cell-Dyn 1800 instrument was studied over the short term (with fresh blood) as well as the long term with stabilized quality control material. In either case, the instrument exhibited satisfactory performance. The blood count parameters showed acceptable stability during storage experiments, although in the case of mean platelet volume, the Cell-Dyn 1800 results increased during the first 6 hours of storage, reflecting the morphological changes expected of platelets from EDTA-anticoagulated specimens. PMID- 14521322 TI - Inclusion of Laboratory Hematology as an indexed journal in Medline. PMID- 14521323 TI - Validation of body fluid analysis on the Coulter LH 750. AB - The role of the hematology laboratory in the analysis of body fluid has been to provide accurate enumeration of red blood cells (RBCs), total nucleated cells (TNCs), and differentials by manual analysis. Three hospitals (London Health Sciences Centre, University of Pittsburgh Medical Center, and University of Michigan Health System) participated in the assessment of the performance of automated analysis of body fluid by the Beckman Coulter LH 750, an impedance based hematology analyzer. We evaluated the accuracy of analysis results for both the TNCs and RBCs of 372 samples (158 serous fluid, 148 cerebrospinal fluid [CSF], 66 synovial fluid) run on the LH 750 compared to results obtained from manual chamber counting. Of the 372 samples, 152 were suitable for evaluation of accuracy of the automated TNC. The remaining 220 samples were either flagged for interfering substances or the reference results were < 0.2 x 10(9)/L, below the background limit of the analyzer. Correlation coefficients for serous fluid were 0.895, P = .88; for CSF, 0.993, P = .84; and for synovial fluid, 0.836, P = .94. Of the 372 samples, 106 had RBC counts greater than 0.01 x 10(12)/L and were used for method comparison. Correlation coefficients for serous fluid were 0.957, P = .66; for CSF, 0.849, P = .55; and for synovial fluid, 0.667, P = .81. Linearity and precision studies showed excellent agreement for both TNC and RBC parameters. Low-level sensitivity excluded the majority of cerebrospinal (119) and a small number of peritoneal dialysate fluid samples (8), which require accurate enumeration at clinical decision points between 0 to 100 cells/microliter. In the case of synovial and serous fluids, however, most clinicians are interested in TNC counts above 0.2 x 10(9)/L, and RBC counts are relevant only if they are significantly increased (> or = 0.05 x 10(12)/L). Adopting the criteria of reporting TNC counts as < 0.2 x 10(9)/L or accurate enumeration on counts > or = 0.2 x 10(9)/L, clinically relevant results could be provided by automated analysis in 93.8% of serous fluids and 85.8% of synovial fluids. PMID- 14521324 TI - Comparison of the AcT 5 diff autoloader hematology analyzer to the Abbott Cell Dyn 3200 analyzer at Charlevoix Area Hospital. AB - The Coulter AcT 5 diff autoloader (AL) hematology analyzer (Beckman Coulter, Fullerton, CA, USA) was evaluated at the Charlevoix Area Hospital, a small rural community hospital. The analyzer was compared to the laboratory's current instrument, a Cell-Dyn 3200 (Abbott Laboratories, Abbott Park, IL, USA). The study included comparisons of precision, efficiency and productivity, and reliability. The 5-part white blood cell differentials on both instruments were compared, and the clinical utility and flagging performance of both analyzers were confirmed by manual differential counts. The AcT 5 diff AL was shown to have excellent precision and accuracy. The addition of the autoloader to the AcT 5 diff provides a low-volume system with the added benefits of a "load and leave" analyzer normally available only to larger laboratories. PMID- 14521325 TI - Evaluation of the Beckman Coulter AcT 5 diff AL hematology analyzer in a hospital setting. AB - The Coulter AcT 5-part differential (5 diff) autoloader (AL) hematology analyzer from Beckman Coulter (Fullerton, CA, USA) was evaluated at the Florida Medical Center in Fort Lauderdale, Florida, an acute care hospital facility. The AcT 5 diff AL is a new, fully automated bench-top 5-part differential hematology analyzer with automatic loading and sampling. It is designed as a front-line instrument for small- to medium-sized laboratories or as a backup in larger laboratories. We evaluated the performance of the AcT 5 diff AL for complete blood counts (CBCs). The Coulter Onyx AL was used as the reference instrument and the 5-part differential against the reference 400-cell manual differential count (2 operators x 200 cells). A total of 140 patient samples were analyzed in the study, with most of the samples obtained from our own laboratory workload. We supplemented this dataset with pediatric and oncology samples from nearby practices. All samples were analyzed within 4 hours of collection, and the AcT 5 diff AL demonstrated excellent accuracy with correlation values (r) > 0.98 for all directly measured CBC parameters (white blood cell count, red blood cell count, hemoglobin, mean corpuscular volume, and platelets). Reproducibility studies on the AcT 5 diff AL also showed excellent results, with coefficients of variation much lower than the manufacturer's specifications. Initial differential comparisons showed good correlations for neutrophils, lymphocytes, and eosinophils (r = 0.84, 0.96, and 0.88, respectively) with a lower correlation for monocytes (r = 0.43). On subsequent review and data analysis we found differential discrepancies in 39 of the 140 samples, for which all showed multiple morphological flags, R diff flags, or poor scatterplot separations indicating that the AcT 5 diff AL differential was likely to be inaccurate and that a morphological assessment and differential was necessary. Given the acutecare setting that our laboratory serves and the skewing of the dataset of our study because of the addition of abnormal pediatric and oncology cases, these abnormal sample results were not surprising. These 39 samples all had markedly abnormal differentials with either increased numbers of immature and/or abnormal cells due to such conditions as end-stage liver disease, respiratory failure, renal failure, sepsis and pneumonia, high white blood cell leukemias, and a variety of other acute illnesses. Reanalysis of the differential data, excluding these 39 grossly abnormal samples with R flags and multiple flags, showed the AcT 5 diff AL differential gave very good correlations for neutrophils, lymphocytes, monocytes, and eosinophils (r = 0.93, 0.97, 0.66, and 0.85, respectively). Our technologists were also impressed with the reliability of the AcT 5 diff AL, its ease of use and transfer from stand-by to ready mode, the user friendliness of the software, and the clarity of the printed report. We concluded that the AcT 5 diff AL gives accurate and precise CBC and 5-part differential results and is an excellent, easy-to-use instrument ideally suited for small- to moderate-sized hospital laboratories, as a back-up in a large hospital laboratory, or for use in physician office laboratories. PMID- 14521326 TI - External quality assessment of automated hematology analyzer performance using fresh human blood samples in Shanghai. AB - We set out to assess the performance and degree of agreement for the different automated hematology analyzers (AHAs) currently used within the Shanghai region. Analyzers from 5 different manufacturers were represented within the 114 hospitals that participated in this project: Abbott, ABX, Beckman Coulter, Nihon Kohden, and Sysmex. We determined the results obtained on 115 hematology analyzers in the 114 different hospitals for hemoglobin (HGB), red blood cell count (RBC), white blood cell count (WBC), hematocrit (HCT), and platelet count (PLT) from fresh anticoagulated blood samples from healthy donors. The maximum coefficients of variation (CV%) among instruments of the 3 main manufacturers (Abbott, Beckman Coulter, and Sysmex) for RBC, HGB, HCT, WBC, and PLT were 3.2%, 3.8%, 3.6%, 9.3%, and 10.8%, respectively. The maximum deviations observed among these parameters from the 5 major instrument groups were 0.74% (RBC), 2.24% (HGB), 6.37% (HCT), 6.97% (WBC), and 21.06% (PLT). Implementation of the regular, local external quality assessment program in the Shanghai region using fresh blood samples has resulted in a reduction of CV% values for the major clinically important CBC parameters (RBC, HGB, HCT, WBC, and PLT) within the instrument groups over time. An important result was that the CBC parameter CVs were also well within established US Clinical Laboratory Improvement Amendments of 1988 guidelines for AHA performance. Combined with a comprehensive education program, the Shanghai external fresh blood quality assessment program has resulted in closer agreement among the various AHA manufacturers, with only very small differences for RBC, HGB, HCT, WBC, and PLT counts. The success of this program has given us increased confidence in the comparability of results and subsequent patient management using the various types of AHAs currently in use in the hospitals within the Shanghai region. PMID- 14521327 TI - A new method for measuring hemoglobin. PMID- 14521328 TI - Probing the mechanistic consequences of 5-fluorine substitution on cytidine nucleotide analogue incorporation by HIV-1 reverse transcriptase. AB - Beta-D and beta-L-enantiomers of 2',3'-dideoxycytidine analogues are potent chain terminators and antimetabolites for viral and cellular replication. Seemingly small modifications markedly alter their antiviral and toxicity patterns. This review discusses previously published and recently obtained data on the effects of 5- and 2'-fluorine substitution on the pre-steady state incorporation of 2' deoxycytidine-5'-monophosphate analogues by HIV-1 reverse transcriptase (RT) in light of their biological activity. The addition of fluorine at the 5-position of the pyrimidine ring altered the kinetic parameters for all nucleotides tested. Only the 5-fluorine substitution of the clinically relevant nucleosides (-)-beta L-2',3'-dideoxy-3'-thia-5-fluorocytidine (L-FTC, Emtriva), and (+)-beta-D-2',3' didehydro-2',3'-dideoxy-5-fluorocytidine (D-D4FC, Reverset), caused a higher overall efficiency of nucleotide incorporation during both DNA- and RNA-directed synthesis. Enhanced incorporation by RT may in part explain the potency of these nucleosides against HIV-1. In other cases, a lack of correlation between RT incorporation in enzymatic assays and antiviral activity in cell culture illustrates the importance of other cellular factors in defining antiviral potency. The substitution of fluorine at the 2' position of the deoxyribose ring negatively affects incorporation by RT indicating the steric gate of RT can detect electrostatic perturbations. Intriguing results pertaining to drug resistance have led to a better understanding of HIV-1 RT resistance mechanisms. These insights serve as a basis for understanding the mechanism of action for nucleoside analogues and, coupled with studies on other key enzymes, may lead to the more effective use of fluorine to enhance the potency and selectivity of antiviral agents. PMID- 14521329 TI - Synthesis, solution conformation and anti-HIV activity of novel 3'-substituted 2',3'-dideoxy-5-hydroxymethyluridines and their 4,5-substituted analogues. AB - To decrease the toxicity of potent anti-HIV nucleosides 3'-azido-2',3' dideoxythymidine (AZT) and 2',3'-dideoxy-3'-fluorothymidine (3'-FddThd, FLT), their new analogues, 3'-azido-2',3'-dideoxy-5-hydroxymethyluridine (3' Az5HmddUrd) and 2',3'-dideoxy-3'-fluoro-5-hydroxymethyluridine (3'-F5HmddUrd), were synthesized. The reaction of 3'-azido-2',3'-dideoxyuridine (3'-AzddUrd) and 2',3'-dideoxy-3'-fluorouridine (3'-FddUrd) with formaldehyde, under strongly alkaline conditions and at elevated temperature, proceeded after 4 days to completion to afford the corresponding 5-hydroxymethyl derivatives 3'-Az5HmddUrd and 3'-F5HmddUrd in good yield. These compounds were also prepared by oxidation of AZT and FLT with the use of K2S2O8. 1H NMR analyses were subjected to the series of 3', 4 and 5-substituted pyrimidine 2'-deoxy- and 2',3' dideoxynucleosides involving 3'-Az5HmddUrd and 3'-F5HmddUrd. Analysis of the sugar furanose ring puckering demonstrated that all 3'-fluorine derivatives exhibited strong domination of the S conformation (approximately 100%) while 3' substitution by electron-donating groups, such as NH2, increased population of the N conformation. Experimentally observed substituent effect on the furanose ring puckering equilibrium was reconstructed in the 100 ps molecular dynamic trajectories obtained for AZT, FLT, dThd, 2',3'-ddThd and 3'-amino-2',3'-ddThd. It may be concluded that anti-HIV activity is linked to a direct interaction of the 3'-substituent with reverse transcriptase (RT) binding site. Anti-HIV activities of 3'-Az5HmddUrd and 3'-F5HmddUrd are lower than activity of AZT and FLT; however, 3'-Az5HmddUrd and 3'-F5HmddUrd are less toxic than AZT and FLT. PMID- 14521330 TI - Synthesis and antiviral activity of some 5'-N-phthaloyl-3'-azido-2',3' dideoxythymidine analogues. AB - A variety of substituted 5'-N-phthaloyl-3'-azido-2',3'-dideoxythymidine derivatives has been evaluated for their activity against HIV-1, HIV-2 and Moloney murine sarcoma virus (MSV) in cell culture. Most of the 3'-azido-2',3' dideoxythymidine (AZT, zidovudine) derivatives showed antiviral activity in the lower micromolar concentration range and there was a close correlation between their anti-HIV and anti-MSV activity (r = 0.99). The adamantyl phthaloyl derivative was active at submicromolar concentrations. None of the compounds showed marked cytostatic activity. They did not inhibit recombinant HIV-1 reverse transcriptase. All compounds were inactive against HIV in thymidine kinase deficient cells, pointing to the compounds' requirement to release free AZT to afford antiviral efficacy. PMID- 14521332 TI - Are the 2-isomers of the drug rimantadine active anti-influenza A agents? AB - There is a lack of information in the medical chemistry literature concerning the anti-influenza A activity of the drug rimantadine's 2-isomer (2-rimantadine). We now present results showing that, although 2-adamantanamine (2-amantadine) 3 is only moderately active, some 2-rimantadine analogues are effective anti-influenza A virus agents in vitro. The 2-rimantadine analogues and their spirocyclobutane and spirocyclopentane congeners were synthesized through interesting routes. The 2-rimantadine analogues were 2-4 times more potent than rimantadine 2 against influenza virus A H2N2 strain; their spirocyclobutane congeners proved equally active to rimantadine 2. Two compounds exhibited a similar activity and one of the compounds was was fourfold more potent than rimantadine 2 against H3N2 strain. PMID- 14521331 TI - Viramidine, a prodrug of ribavirin, shows better liver-targeting properties and safety profiles than ribavirin in animals. AB - Ribavirin, part of the current first line combination therapy for the treatment of chronic hepatitis C, may cause haemolytic anaemia and poses a significant challenge to the clinical management of the disease. Viramidine, a prodrug of ribavirin, is currently under development. In-vitro partition demonstrated that viramidine had less association with RBCs than ribavirin in rat, monkey and man, and thus has less liability for haemolytic anaemia than ribavirin. In a whole body autoradiography study in rats following oral dosing (30 mg/kg) of [14C]ribavirin or [14C]viramidine to monkeys, viramidine produced 32% higher radioactivity in the liver than ribavirin, indicating a better liver-targeting properties. In portal vein-cannulated cynomolgus monkeys following single oral dosing (30 mg/kg) of [3H]viramidine or [3H]ribavirin, viramidine retained 3X higher radioactivity in the liver than ribavirin. Viramidine dosing also produced a higher viramidine to ribavirin ratio in portal plasma than in systemic plasma, indicating that the liver was the main site for the viramidine conversion to ribavirin and subsequent trapping of the drug. After multiple oral dosing (10 mg/kg) of [14C]ribavirin or [14C]viramidine to monkey, viramidine yielded three times the drug level in the liver but only half in RBCs compared to ribavirin. Viramidine and ribavirin had comparable toxicity profiles in a 28-day toxicity study in rats. In contrast, viramidine had much better safety profiles than ribavirin in a 28-day toxicity study in monkeys. In conclusion, viramidine has better liver-targeting properties and safety profiles than ribavirin in animals. PMID- 14521333 TI - Halophenyl furanopyrimidines as potent and selective anti-VZV agents. AB - Bicyclic furano pyrimidines have been previously reported by us to be highly potent and selective inhibitors of varicella zoster virus (VZV). p-Alkyl phenyl analogues are particularly potent with EC50 values below 1 nM. In this article we report the synthesis and anti-VZV activity of a series of halophenyl analogues, with variation in the nature (F, Cl, Br) and location (o, m, p) of the halogen substituent. The compounds show a range of activities from ca. 10 nM to > 50 microM. In most cases, ortho substitution leads to greatest activity, meta substitution is in general poor, and the effect of p-substitution shows a marked dependence on the halogen atom. The p-fluorophenyl compound is unique amongst compounds of this class in being inactive as an antiviral. The possible origins of these marked SARs are discussed. PMID- 14521400 TI - 3,6-dibromocarbazole piperazine derivatives of 2-propanol as first inhibitors of cytochrome c release via Bax channel modulation. AB - There is compelling evidence that Bax channel activity stimulates cytochrome c release leading ultimately to cell death, which is a key event in ischemic injuries and neurodegenerative diseases. Here 3,6-dibromocarbazole piperazine derivatives of 2-propanol are described as the first small and potent modulators of the cytochrome c release triggered by Bid-induced Bax activation in a mitochondrial assay. Furthermore, a mechanism of action is proposed, and fluorescent derivatives allowing the localization of such inhibitors are reported. PMID- 14521401 TI - Facile and selective nanoscale labeling of peptides in solution by using photolabile protecting groups. AB - For the selective labeling of peptides, a novel strategy was developed that combines the advantages of solid-phase peptide synthesis with the flexibility of labeling reactions in solution. To direct a label at a distinct position within the peptide sequence, other reactive positions are blocked with photolabile protecting groups that could be easily removed after the labeling reaction. Therefore selective labeling may become feasible for the first time even in nanomol amounts. PMID- 14521402 TI - Esters, retroesters, and a retroamide of palmitic acid: pool for the first selective inhibitors of N-palmitoylethanolamine-selective acid amidase. AB - Cyclohexyl hexadecanoate, hexadecyl propionate, and N-(3 hydroxypropionyl)pentadecanamide, respectively ester, retroester, and retroamide derivatives of N-palmitoylethanolamine, represent the first selective inhibitors of "N-palmitoylethanolamine hydrolase" described so far. These compounds are devoid of affinity for CB(1) and CB(2) receptors and characterized by high percentages of inhibition of N-palmitoylethanolamine-selective acid amidase (84.0, 70.5, and 76.7% inhibition at 100 microM, respectively) with much lower inhibitory effect on either fatty acid amide hydrolase or the uptake of anandamide. PMID- 14521404 TI - Docking studies on alphavbeta3 integrin ligands: pharmacophore refinement and implications for drug design. AB - Starting from the first crystal structure of the extracellular segment of the alpha(v)beta(3) integrin receptor with a cyclic RGD ligand bound to the active site, structural models for the interactions of known ligands with the alpha(v)beta(3) integrin receptor were generated by automated computational docking. The obtained complexes were evaluated for their consistency with structure-activity relationships and site-directed mutagenesis data. A comparison between the calculated interaction free energies and the experimental biological activities was also made. All the possible interactions of the investigated compounds at the active site and the probable ligand binding conformations provide an improved basis for structure-based rational ligand design. Additionally, our docking results allow a further validation and refinement of the pharmacophore model previously postulated by us. PMID- 14521403 TI - Molecular modeling of the three-dimensional structure of dopamine 3 (D3) subtype receptor: discovery of novel and potent D3 ligands through a hybrid pharmacophore and structure-based database searching approach. AB - The dopamine 3 (D3) subtype receptor has been implicated in several neurological conditions, and potent and selective D3 ligands may have therapeutic potential for the treatment of drug addiction, Parkinson's disease, and schizophrenia. In this paper, we report computational homology modeling of the D3 receptor based upon the high-resolution X-ray structure of rhodopsin, extensive structural refinement in the presence of explicit lipid bilayer and water environment, and validation of the refined D3 structural models using experimental data. We further describe the development, validation, and application of a hybrid computational screening approach for the discovery of several classes of novel and potent D3 ligands. This computational approach employs stepwise pharmacophore and structure-based searching of a large three-dimensional chemical database for the identification of potential D3 ligands. The obtained hits are then subjected to structural novelty screening, and the most promising compounds are tested in a D3 binding assay. Using this approach we identified four compounds with K(i) values better than 100 nM and eight compounds with K(i) values better than 1 microM out of 20 compounds selected for testing in the D3 receptor binding assay. Our results suggest that the D3 structural models obtained from this study may be useful for the discovery and design of novel and potent D3 ligands. Furthermore, the employed hybrid approach may be more effective for lead discovery from a large chemical database than either pharmacophore-based or structure-based database screening alone. PMID- 14521405 TI - Structure-based design of novel guanidine/benzamidine mimics: potent and orally bioavailable factor Xa inhibitors as novel anticoagulants. AB - As part of an ongoing effort to prepare orally active factor Xa inhibitors using structure-based drug design techniques and molecular recognition principles, a systematic study has been performed on the pharmacokinetic profile resulting from replacing the benzamidine in the P1 position with less basic benzamidine mimics or neutral residues. It is demonstrated that lowering the pK(a) of the P1 ligand resulted in compounds (3-benzylamine, 15a; 1-aminoisoquinoline, 24a; 3 aminobenzisoxazole, 23a; 3-phenylcarboxamide, 22b; and 4-methoxyphenyl, 22a) with improved pharmacokinetic features mainly as a result of decreased clearance, increased volume of distribution, and enhanced oral absorption. This work resulted in a series of potent and orally bioavailable factor Xa inhibitors that ultimately led to the discovery of SQ311, 24a. SQ311, which utilizes a 1 aminoisoquinoline as the P1 ligand, inhibits factor Xa with a K(i) of 0.33 nM and demonstrates both good in vivo antithrombotic efficacy and oral bioavailability. PMID- 14521406 TI - Understanding the antitumor activity of novel hydroxysemicarbazide derivatives as ribonucleotide reductase inhibitors using CoMFA and CoMSIA. AB - Three-dimensional quantitative structure-activity relationship (3D-QSAR) studies were performed on a series of Schiff bases of hydroxysemicarbazide analogues using comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) methods with their antitumor activities against L1210 cells. The models were generated using 24 molecules, out of which one molecule was a commercially available ribonucleotide reductase (RR) inhibitor, hydroxyurea (HU), and the predictive ability of the resulting each model was evaluated against a test set of four molecules. Maximum common substructure (MCS)-based method was used for alignment and compared with the known alignment methods. The QSAR models from both methods exhibited considerable correlative and predictive properties. Inclusion of additional descriptor ClogP improved the statistics of CoMFA model significantly. Both methods strongly suggest the necessity of lipophilicity for antitumor activity. CoMFA and CoMSIA methods predicted HU optimally, indicating a similar mechanism of action for the molecules considered for generating the models and HU to inhibit the tumor cells. The analysis of CoMFA contour maps provided insight into the possible modification of the molecules for better activity. PMID- 14521407 TI - Design and synthesis of pyrrolidine-5,5'-trans-lactams (5-oxo hexahydropyrrolo[3,2-b]pyrroles) as novel mechanism-based inhibitors of human cytomegalovirus protease. 4. Antiviral activity and plasma stability. AB - A series of chiral, (S)-proline-alpha-methylpyrrolidine-5,5-trans-lactam serine protease inhibitors has been developed as antivirals of human cytomegalovirus (HCMV). The SAR of the functionality on the proline nitrogen has shown that derivatives of para-substituted phenyl ureas > para-substituted phenyl sulfonamides > para-substituted phenyl carboxamide for activity against HCMV deltaAla protease, producing para-substituted phenyl ureas with single figure nM potency (K(i)) against the viral enzyme. The SAR of the functionality on the lactam nitrogen has defined the steric and electronic requirements for high human plasma stability while retaining good activity against HCMV protease. The combination of high potency against HCMV deltaAla protease and high human plasma stability has produced compounds with significant in vitro antiviral activity against human cytomegalovirus with the 6-hydroxymethyl benzothiazole derivative 72 being equivalent in potency to ganciclovir. The parent benzothiazole 56 had good pharmacokinetics in dogs with 29% bioavailability and good brain and ocular penetration in guinea pigs. PMID- 14521408 TI - Three-dimensional models for beta-adrenergic receptor complexes with agonists and antagonists. AB - Molecular modeling methods have been used to construct three-dimensional models for agonist and antagonist complexes with beta-adrenergic receptors. The recent rhodopsin crystal structure was used as a template in standard homology modeling methods. The rhodopsin-based homology models were assessed for agreement with experimental results for beta-adrenergic receptors, and compared with receptor models developed using de novo modeling techniques. While the de novo and homology-derived receptor models are generally quite similar, there are some localized structural differences that impact the putative ligand-binding site significantly. The de novo receptor models appear to provide much better agreement with experimental data, particularly for receptor complexes with agonist ligands. The de novo receptor models also yield some interesting and testable hypotheses for the structural basis of beta-adrenergic receptor subtype ligand selectivity. PMID- 14521409 TI - Trisubstituted acridine derivatives as potent and selective telomerase inhibitors. AB - The synthesis and evaluation for telomerase-inhibitory and quadruplex DNA binding properties of three related series of rationally designed trisubstituted acridine derivatives are described. These are substituted on the acridine ring at the 2,6,9; 2,7,9; and 3,6,9 positions. The ability of several of the most potent compounds to interact with and stabilize an intramolecular G-quadruplex DNA was evaluated by surface plasmon resonance methods, and affinities were found to correlate with potency in a telomerase assay. The interactions of a number of compounds with a parallel quadruplex DNA structure were simulated by molecular modeling methods. The calculated interaction energies were compared with telomerase activity and showed generally consistent correlations between quadruplex affinity and telomerase inhibition. These data support a model for the action of these compounds that involves the stabilization of intermediate quadruplex structures that inhibit the elongation of telomeric DNA by telomerase in tumor cells. PMID- 14521410 TI - Identification and prediction of promiscuous aggregating inhibitors among known drugs. AB - Some small molecules, often hits from screening, form aggregates in solution that inhibit many enzymes. In contrast, drugs are thought to act specifically. To investigate this assumption, 50 unrelated drugs were tested for promiscuous inhibition via aggregation. Each drug was tested against three unrelated model enzymes: beta-lactamase, chymotrypsin, and malate dehydrogenase, none of which are considered targets of these drugs. To be judged promiscuous, the drugs had to inhibit all three enzymes, do so in a time-dependent manner, be sensitive to detergent and to enzyme concentration, and form particles detectable by light scattering. Of the 50 drugs tested, 43 were nonpromiscuous by these criteria. Surprisingly, four of the drugs showed promiscuous, aggregation-based inhibition at concentrations below 100 microM: clotrimazole, benzyl benzoate, nicardipine, and delavirdine. Three other drugs also behaved as aggregation-based inhibitors, but only at high concentrations (about 400 microM). To investigate possible structure-activity relationships among promiscuous drugs, five analogues of the antifungal clotrimazole were studied. Three of these, miconazole, econazole, and sulconazole, were promiscuous but the other two, fluconazole and ketoconazole, were not. Using recursive partitioning, these experimental results were used to develop a model for predicting aggregate-based promiscuity. This model correctly classified 94% of 111 compounds-47 aggregators and 64 nonaggregators-that have been studied for this effect. To evaluate the model, it was used to predict the behavior of 75 drugs not previously investigated for aggregation. Several preliminary points emerge. Most drugs are not promiscuous, even at high concentrations. Nevertheless, at high enough concentrations (20-400 microM), some drugs can aggregate and act promiscuously, suggesting that aggregation may be common among small molecules at micromolar concentrations, at least in biochemical buffers. PMID- 14521411 TI - Simple, intuitive calculations of free energy of binding for protein-ligand complexes. 2. Computational titration and pH effects in molecular models of neuraminidase-inhibitor complexes. AB - One factor that can strongly influence predicted free energy of binding is the ionization state of functional groups on the ligands and at the binding site at which calculations are performed. This analysis is seldom performed except in very detailed computational simulations. In this work, we address the issues of (i) modeling the complexity resulting from the different ionization states of ligand and protein residues involved in binding, (ii) if, and how, computational methods can evaluate the pH dependence of ligand inhibition constants, and (iii) how to score the protonation-dependent models. We developed a new and fairly rapid protocol called "computational titration" that enables parallel modeling of multiple ionization ensembles for each distinct protonation level. Models for possible protonation combinations for site/ligand ionizable groups are built, and the free energy of interaction for each of them is quantified by the HINT (Hydropathic INTeractions) software. We applied this procedure to the evaluation of the binding affinity of nine inhibitors (six derived from 2,3-didehydro-2 deoxy-N-acetylneuraminic acid, DANA) of influenza virus neuraminidase (NA), a surface glycoprotein essential for virus replication and thus a pharmaceutically relevant target for the design of anti-influenza drugs. The three-dimensional structures of the NA enzyme-inhibitor complexes indicate considerable complexity as the ligand-protein recognition site contains several ionizable moieties. Each computational titration experiment reveals a peak HINT score as a function of added protons. This maximum HINT score indicates the optimum pH (or the optimum protonation state of each inhibitor-protein binding site) for binding. The pH at which inhibition is measured and/or crystals were grown and analyzed can vary from this optimum. A protonation model is proposed for each ligand that reconciles the experimental complex structure with measured inhibition and the free energy of binding. Computational titration methods allow us to analyze the effect of pH in silico and may be helpful in improving ligand binding free energy prediction when protonation or deprotonation of the residues or ligand functional groups at the binding site might be significant. PMID- 14521412 TI - Generation of predictive pharmacophore models for CCR5 antagonists: study with piperidine- and piperazine-based compounds as a new class of HIV-1 entry inhibitors. AB - Predictive pharmacophore models were developed for a large series of piperidine- and piperazine-based CCR5 antagonists as anti-HIV-1 agents reported by Schering Plough Research Institute in recent years. The pharmacophore models were generated using a training set consisting of 25 carefully selected antagonists based on well documented criteria. The activity spread, expressed in K(i), of training set molecules was from 0.1 to 1300 nM. The most predictive pharmacophore model (hypothesis 1), consisting of five features, namely, two hydrogen bond acceptors and three hydrophobic, had a correlation (r) of 0.920 and a root mean square of 0.879, and the cost difference between null cost and fixed cost was 44.46 bits. The model was cross-validated by randomizing the data using the CatScramble technique. The results confirmed that the pharmacophore models generated from the test set were not due to chance correlation. The best model (hypothesis 1) was validated using test set molecules (total of 78) and performed well in classifying active and inactive molecules correctly. The model was further validated by mapping onto it a diverse set of six CCR5 antagonists identified by five different pharmaceutical companies. The best model correctly predicted these compounds as being highly active. These multiple validation approaches provide confidence in the utility of the predictive pharmacophore model developed in this study as a 3D query tool in virtual screening to retrieve new chemical entities as potent CCR5 antagonists. The model can also be used in predicting biological activities of compounds prior to undertaking their costly synthesis. PMID- 14521413 TI - Novel derivatives of benzo[b]thieno[2,3-c]quinolones: synthesis, photochemical synthesis, and antitumor evaluation. AB - Novel derivatives of benzo[b]thieno[2,3-c]quinolones 3a-j were synthesized in a multistep synthesis starting from substituted benzo[b]thiophene-2-carbonyl chlorides, to their corresponding benzo[b]thiophene-2-carboxamides, which were photochemically dehydrohalogenated to their corresponding substituted benzo[b]thieno[2,3-c]quinolones. Compound 4 was prepared from 3i by alkylation with 3-dimethylaminopropyl chloride in the presence of NaH. Compounds 7a,b were prepared from 3g in the multistep synthesis from compounds 5 and 6. Compounds 3b, 3c-f, 3h, 7a, and 7b were found to exert cytostatic activity against malignant cell lines: pancreatic carcinoma (MiaPaCa2), breast carcinoma (MCF7), cervical carcinoma (HeLa), laryngeal carcinoma (Hep2), colon carcinoma (CaCo-2), melanoma (HBL), human fibroblast cell lines (WI-38). The compounds that bear a 3 dimethylaminopropyl substituent on the quinolone nitrogen (3b, 3c-f, 3h) showed higher antitumor activity than compounds bearing the same substituent on the amidic nitrogen (7a and 7b). The compound 3h, which has a 3-dimethylaminopropyl substituent on the quinolone nitrogen and a methoxycarbonyl substituent at position 9, had marked antitumor activity. Because of strong cytotoxic effect of compound 4 on melanoma cells (HBL, ME 67.3, and ME 67.1), a potential mechanism of action was examined. Analysis of DNA and Annexin-V-FLUOS staining indicated that compound 4 causes cell death by apoptosis. PMID- 14521414 TI - The discovery of tadalafil: a novel and highly selective PDE5 inhibitor. 1: 5,6,11,11a-tetrahydro-1H-imidazo[1',5':1,6]pyrido[3,4-b]indole-1,3(2H)-dione analogues. AB - Starting from ethyl beta-carboline-3-carboxylate (beta-CCE), 1, a modest inhibitor of type 5 phosphodiesterase (PDE5), a series of functionalized tetrahydro-beta-carboline derivatives has been identified as a novel chemical class of potent and selective PDE5 inhibitors. Optimization of the side chain on the hydantoin ring of initial lead compound 2 and of the aromatic ring on position 5 led to the identification of compound 6e, a highly potent and selective PDE5 inhibitor, with greater selectivity for PDE5 vs PDE1-4 than sildenafil. Compound 6e demonstrated a long-lasting and significant blood pressure lowering effect after iv administration in the spontaneously hypertensive rat model but showed only moderate oral in vivo efficacy. PMID- 14521415 TI - The discovery of tadalafil: a novel and highly selective PDE5 inhibitor. 2: 2,3,6,7,12,12a-hexahydropyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione analogues. AB - Modification of the hydantoin ring in the previously described lead compound 2a has led to the discovery of compound 12a, tadalafil, a highly potent and highly selective PDE5 inhibitor. The replacement of the hydantoin in compound 2a by a piperazinedione ring led to compound cis-11a which showed similar PDE5 inhibitory potency. Introduction of a 3,4-methylenedioxy substitution on the phenyl ring in position 6 led to a potent PDE5 inhibitor cis-11c with increased cellular potency. Optimization of the chain on the piperazinedione ring led to the identification of the racemic cis-N-methyl derivative 11i. High diastereospecificity for PDE5 inhibition was observed in the piperazinedione series with the cis-(6R,12aR) enantiomer displaying the highest PDE5 inhibitory activity. The piperazinedione 12a, tadalafil (GF196960), has been identified as a highly potent PDE5 inhibitor (IC(50) = 5 nM) with high selectivity for PDE5 vs PDE1-4 and PDE6. Compound 12a displays 85-fold greater selectivity vs PDE6 than sildenafil 1. 12a showed profound and long-lasting blood pressure lowering activity (30 mmHg/>7 h) in the spontaneously hypertensive rat model after oral administration (5 mg/kg). PMID- 14521416 TI - New prolyl oligopeptidase inhibitors developed from dicarboxylic acid bis(l prolyl-pyrrolidine) amides. AB - Isophthalic acid bis(l-prolyl-pyrrolidine) amide is a very potent prolyl oligopeptidase inhibitor, but it has a log P value of -0.2, which is very low for a compound targeted to the brain. Therefore, these types of compounds were further modified to improve the structure-activity relationships, with the focus on increasing the log P value. The inhibitory activity against prolyl oligopeptidase from pig brain was tested in vitro. The most promising compounds resulted from replacing the pyrrolidinyl group at the P5 site by cycloalkyl groups, such as cyclopentyl and cyclohexyl groups, and by a phenyl group. These compounds are slightly more potent, and they have a significantly higher log P value. The potency of these compounds was further increased by replacing the pyrrolidinyl group at the P1 site by 2(S)-cyanopyrrolidinyl and 2(S) (hydroxyacetyl)pyrrolidinyl groups. PMID- 14521417 TI - Design, synthesis, and anticancer properties of 4,4'-dihydroxybenzophenone-2,4 dinitrophenylhydrazone and analogues. AB - 4,4'-dihydroxybenzophenone-2,4-dinitrophenylhydrazone (A-007) has recently completed a phase I clinical trial in advanced cancer with minimal toxicity, and impressive objective responses were noted. A-007 possesses three moieties that appear to have an influence on its anticancer activities: diphenylmethane, hydrazone, and dinitrophenyl. The goals of this study were to modify A-007's chemical moieties with the ultimate goal of maximizing its anticancer activity through increased planarity and introduction of functional groups. Thirty-five phenylhydrazone analogues of A-007 were synthesized and evaluated in vitro in a human primary cancer explant assay. Anticancer activities for selected analogues were also assayed for activity vs established human/murine cell lines. One hundred-eighty-six fresh human solid tumors were used to screen for anticancer activity. Selected analogues were assayed for therapeutic indices (vs GM-CFC from bone marrow) in preparation for preclinical studies. Several polyaryl phenylhydrazones demonstrated improved cytotoxic activities by factors of 10(2) 10(3) when compared with A-007. However, the polyaryl quinone moieties of the latter analogues introduced potential toxic properties (cardiac, hematological) that do not exist with A-007. PMID- 14521418 TI - S-acyl-2-thioethyl phosphoramidate diester derivatives as mononucleotide prodrugs. AB - The synthesis and in vitro anti-HIV activity of phosphoramidate diester derivatives of 3'-azido-2',3'-dideoxythymidine (AZT) bearing one S-pivaloyl-2 thioethyl (tBuSATE) group and various amino residues are reported. These compounds were obtained from an H-phosphonate strategy using an amidative oxidation step. Most of these derivatives appeared to inhibit HIV-1 replication, with EC(50) values at micromolar concentration in thymidine kinase-deficient (TK ) cells, revealing a less restrictive intracellular decomposition process than previously reported for other phosphoramidate prodrugs. The proposed decomposition pathway of this new series of mixed pronucleotides may successively involve an esterase and a phosphoramidase hydrolysis. PMID- 14521420 TI - Cyclopropane-containing polyamine analogues are efficient growth inhibitors of a human prostate tumor xenograft in nude mice. AB - Polyamine analogues 7, 10, 18, 27, and 32 containing cyclopropane rings were obtained by chemical synthesis. Their antineoplastic activities were assessed against the cultured human prostate tumor cell lines DU-145, DuPro, and PC-3. Decamines 32 and 27 exhibited variable levels of cytotoxicity against all three cell lines, while 7, 10, and 18 were efficacious against DU-145 and DuPro. Maximum tolerated doses (MTD) for all five compounds in a NCr-nu mouse model were determined at dosing schedules of q1d x 5 (ip) in two cycles with a break of 10 days between cycles. Their antitumor efficacies were then tested against DU-145 tumor xenografts in mice treated with all five agents at their respective MTDs. In addition, the efficacies of 7 and 10 against the same tumor xenograft were assessed at doses below their respective MTDs. In all experiments, administration began two weeks after tumor implantation. All compounds efficiently inhibited tumor growth for up to 50 days postimplantation, with negligible animal body weight loss. Tetramine 10 and hexamine 18 were the most efficient among the five analogues in arresting tumor growth. Tetramine 10 containing two cyclopropane rings had the lowest systemic toxicity as reflected in animal body weight loss. It was further assessed at a weekly administration regimen of (q1w x 4) in two cycles with a four-week break between the cycles. At this dosing schedule, 10 again efficiently arrested tumor growth with negligible effect on animal body weight. Tetramine 10 also arrested the growth of large tumors (ca. 2000 mm(3)) treated 66 days postimplantation. Studies on the metabolism of 10 showed that it accumulates in tumor within 6 h after the end of administration and reached a maximum level 72 h after cessation of dosing. Intracellular concentrations of 10 in liver and kidney were much smaller when compared to those in the tumor when measured 72 h after cessation of dosing. In liver and kidney, the deethyl metabolites of 10 accumulated over a 96 h period after cessation of dosing. PMID- 14521419 TI - Structure-based design, synthesis, and biological evaluation of irreversible human rhinovirus 3C protease inhibitors. 8. Pharmacological optimization of orally bioavailable 2-pyridone-containing peptidomimetics. AB - The optimization of the pharmacokinetic performance of various 2-pyridone containing human rhinovirus (HRV) 3C protease (3CP) inhibitors following oral administration to either beagle dogs or CM-monkeys is described. The molecules described in this work are composed of a 2-pyridone-containing peptidomimetic binding determinant and an alpha,beta-unsaturated ester Michael acceptor moiety which forms an irreversible covalent adduct with the active site cysteine residue of the 3C enzyme. Modification of the ester contained within these compounds is detailed along with alteration of the P(2) substituent present in the peptidomimetic portion of the inhibitors. The pharmacokinetics of several inhibitors in both dogs and monkeys are described (7 h plasma concentrations after oral administration) along with their human plasma stabilities, stabilities in incubations with human, dog, and monkey microsomes and hepatocytes, Caco-2 permeabilities, and aqueous solubilities. Compounds containing an alpha,beta unsaturated ethyl ester fragment and either an ethyl or propargyl P(2) moiety displayed the most promising combination of 3C enzyme inhibition (k(obs)/[I] 170 000-223 000 M(-1) s(-1)), antiviral activity (EC(50) = 0.047-0.058 microM, mean vs seven HRV serotypes), and pharmacokinetics following oral administration (7 h dog plasma levels = 0.248-0.682 microM; 7 h CM-monkey plasma levels = 0.057-0.896 microM). PMID- 14521421 TI - Probing the interactions of phosphosulfomannans with angiogenic growth factors by surface plasmon resonance. AB - The binding interactions of the phosphosulfomannan anticancer agent PI-88 (1) with the angiogenic growth factors FGF-1, FGF-2, and VEGF were studied by surface plasmon resonance (SPR) on a BIAcore 3000 biosensor. Compared with heparin, PI-88 has at least 11-fold higher affinity for FGF-1 and at least 3-fold higher affinity for VEGF, but at least 13-fold lower affinity for FGF-2. To define the structural features of PI-88 that are important for growth factor binding, several analogues, such as dephosphorylated PI-88 and a sulfated pentasaccharide, were prepared. The binding interactions of these analogues with FGF-1, FGF-2, and VEGF were similarly studied by SPR, and structure-activity relationships were determined. PMID- 14521422 TI - N-hydroxy-3-phenyl-2-propenamides as novel inhibitors of human histone deacetylase with in vivo antitumor activity: discovery of (2E)-N-hydroxy-3-[4 [[(2-hydroxyethyl)[2-(1H-indol-3-yl)ethyl]amino]methyl]phenyl]-2-propenamide (NVP LAQ824). AB - A series of N-hydroxy-3-phenyl-2-propenamides were prepared as novel inhibitors of human histone deacetylase (HDAC). These compounds were potent enzyme inhibitors, having IC(50)s < 400 nM in a partially purified enzyme assay. However, potency in cell growth inhibition assays ranged over 2 orders of magnitude in two human carcinoma cell lines. Selected compounds having cellular IC(50) < 750 nM were tested for maximum tolerated dose (MTD) and for efficacy in the HCT116 human colon tumor xenograft assay. Four compounds having an MTD > or = 100 mg/kg were selected for dose-response studies in the HCT116 xenograft model. One compound, 9 (NVP-LAQ824), had significant dose-related activity in the HCT116 colon and A549 lung tumor models, high MTD, and low gross toxicity. On the basis, in part, of these properties, 9 has entered human clinical trials in 2002. PMID- 14521423 TI - Exposure to noise on board locomotives. AB - Personal and area noise dosimetry measurements were taken in the cabs of leading and trailing locomotives on 48 trips, under winter and summer conditions, on 9 different routes. The mean equivalent sound level (L(EQ), 3 dB exchange rate, 50 dBA threshold) of the engineers and conductors was 84 dBA during winter and 88 dBA during summer. The corresponding time-weighted average levels (L(TWA), 5 dB exchange rate, 80 dBA threshold) were 80 and 84 dBA respectively. The L(EQ) of 56% of the engineers sampled was > or =85 dBA and of 13% was > or =90 dBA. Plots of L(EQ) time history show that under normal operating conditions L(EQ) reaches its steady-state value in about 3 hours. The mean noise levels in the trailing cabs were lower than the personal exposure levels of the engineers and conductors. The mean L(EQ) on the engineer and conductor sides was 80 dBA during winter, and 85 dBA during summer. Locomotive configuration has a significant effect on the noise levels in the trailing cab. The forward-backward configuration resulted in higher noise levels than the forward-forward configuration. Octave and one-third octave band spectra taken during a variety of locomotive operating conditions are presented. The octave band centered at 31.5 Hz contains nearly 46% of the acoustical energy, and those centered at and below 250 Hz contain nearly 99% of the acoustical energy. Wheel-rail interaction appears to be the predominant source of the low frequency noise. Recommendations for controlling exposure are made. PMID- 14521424 TI - Hexavalent chromium exposures during full-aircraft corrosion control. AB - Aluminum alloys used in the construction of modern aircraft are subject to corrosion. The principal means of controlling this corrosion in the U.S. Air Force are organic coatings. The organic coating system consists of a chromate conversion coat, epoxy resin primer, and polyurethane enamel topcoat. Hexavalent chromium (CrVI) is present in the conversion coat in the form of chromic acid and in the primer in the form of strontium chromate. CrVI inhalation exposures can occur when workers spray conversion coat onto bare metal and apply primer to the treated metal surface. In addition, mechanical abrasion of aircraft surfaces can generate particulates that contain chromates from previously applied primers and conversion coats. This study measured CrVI exposures during these corrosion control procedures. Mean time-weighted average (TWA) exposure to chromic acid during conversion coat treatment was 0.48 microg/m(3), below the current American Conference of Governmental Industrial Hygienists (ACGIH) threshold limit value (TLV(R)) TWA of 50 microg/m(3) for water-soluble CrVI compounds. Mean TWA exposures to strontium chromate were 5.33 microg/m(3) during mechanical abrasion and 83.8 microg/m(3) during primer application. These levels are in excess of the current ACGIH TLV-TWA of 0.5 microg/m(3) for strontium chromate. In the absence of a change from chromated to nonchromated conversion coats and primers, additional control measures are needed to reduce these exposures. PMID- 14521425 TI - Farmers' exposure to airborne microorganisms in composting swine confinement buildings. AB - Farmers' exposure to airborne microorganisms was studied in 12 composting swine confinement buildings and in 7 buildings with traditional slatted-floor pit systems. Airborne cultivable mesophilic, xerophilic, and thermotolerant fungi, mesophilic bacteria, and thermophilic actinobacteria were determined with a six stage impactor. Furthermore, the total concentrations of microorganisms were determined with filter sampling and direct count using a microscope. In swine confinement buildings where the composting system was functioning properly, the concentrations of microorganisms were 10-1000 times higher than in traditional swine buildings. High concentrations of thermotolerant fungi and thermophilic actinobacteria (up to 10(5) CFU/m(3)), which have been considered to be the main causative agents of farmer's lung, were found in the composting swine confinement buildings that were studied. The conclusion was reached that farmers are exposed to high concentrations of fungal and actinobacterial spores also in swineries, at least in composting confinement buildings. Therefore, personal protection is strongly recommended in composting swineries, especially during the turning of the compost bed. PMID- 14521426 TI - Exposure to diesel exhaust emissions on board locomotives. AB - Measurements of diesel exhaust emissions (DEEs) were taken in the cabs of leading and trailing locomotives on 48 runs, under winter and summer conditions, on 9 different routes. The cab windows were kept open during the summer runs and closed during the winter runs. The average measurement duration was 9.5 hours. There was virtually no exposure to DEEs in the lead locomotives during winter or summer and very little in the trailing locomotives during winter. The average elemental carbon (EC) concentration in the trailing units of the summer trials was greater than or equal to the proposed American Conference of Governmental Industrial Hygienists' threshold limit value/time-weighted average of 20 microg/m(3) on 26% of the runs, and was greater than or equal to 10 microg/m(3) on 63%. The concentrations of the gaseous components (nitric oxide, nitrogen dioxide, and carbon monoxide) were from 10 to 20 times below their respective threshold limit values. Mean EC concentration was 2.9 microg/m(3) (detection limit 2 microg/m(3)) during the winter runs and 17.1 microg/m(3) during summer. DEEs appeared to be fairly uniformly distributed in the trailing cabs. Configuration of the locomotives had a major impact on EC concentration, with the mean concentration being nearly three times higher in the forward-backward mode than in the forward-forward mode. Descriptive statistics such as means, medians, standard deviations, and so forth, are provided. Various types of statistical comparisons are reported. Recommendations for controlling exposure are made. PMID- 14521427 TI - Using multiple information sources to identify opportunities for ergonomic interventions in automotive parts distribution: a case study. AB - An essential activity in any ergonomics program is determining specific work locations and activities where physical demands place workers at increased risk of sustaining an overexertion injury. To do this, safety and health professionals rely on a variety of information sources to identify and prioritize opportunities for ergonomic interventions. As part of a 4-year project to reduce overexertion injuries in the service parts division of a major auto maker, a study was performed in 19 parts distribution centers to evaluate the contributions of the following information sources in identifying specific high-risk work locations and activities: (1) archival medical/injury records, (2) identification of "problem tasks" by plant-based ergonomic committees, (3) facility walk-throughs by experienced ergonomists, and (4) detailed ergonomic job analyses. Archival records were not particularly useful in identifying high-risk activities because essential exposure information (e.g., task, work location) was not documented. Walk-throughs and detailed ergonomic analyses were partially effective in identifying high-risk activities; however, in some cases the observation time was too short to observe peak exposures. Ergonomic committees were generally effective in identifying specific high-risk tasks and work locations. Rankings of "problem tasks" from multiple sites identified consensus division-wide ergonomic concerns. Detailed ergonomic job analyses confirmed that these tasks had high exposure to ergonomic risk factors. This study demonstrated potential pitfalls in relying on a single information source to identify work locations and activities that place workers at increased risk of overexertion injury. PMID- 14521428 TI - The performance of culture-based methods and microscopy for quantification of noninfectious airborne microorganisms in epidemiological studies of highly contaminated work environments. AB - Airborne levels of microorganisms traditionally have been measured by culture based methods. Culture-based methods are suitable for the detection of infectious agents, but their suitability for the detection of microorganisms with toxic and allergic effects is less clear, because these effects do not depend on viability of the organisms. During the last 15 years several noncultural methods have been developed for the quantification of airborne microorganisms, including microscopic methods. Microscopy may be expected to provide more valid exposure estimates of microorganisms than culture-based methods, because live and dead microorganisms can be detected. However, their validity may also depend on the ability to differentiate between species. The literature was searched for epidemiological studies in which exposure-response analyses were carried out using culture-based methods and/or microscopy. The influence of several factors on exposure-response associations were considered: design; population size; analytical method; sampling method; exposure levels; outcome; and confounder adjustment. Thirteen studies were found, including a total of 49 exposure response analyses, and 45% of the analyses showed associations. It was found that the potential of microscopic methods to uncover exposure-response associations was only marginally better than that of culture-based methods (47 and 44%, respectively). Exposure-response associations were more often found with fungi (70%) than with gram-negative bacteria (50%) or total bacteria (22%), perhaps because fungal exposure is more strongly associated to respiratory outcomes than exposure to bacteria. But the shortcomings of the measurement methods may also be important. Further development of measurement methods for bacteria is therefore needed. The complex composition of bioaerosols in many work environments necessitate the assessment of exposure to multiple agents and multivariate statistical analysis of exposure-response associations. PMID- 14521429 TI - Achieving health, safety, and performance improvements through enhanced cost visibility and workplace partnerships. AB - Reduction in the environment, safety, and occupational health (ESOH) component of operational costs requires not only a better understanding of ESOH costs and requirements, but also the formation of effective partnerships between ESOH professionals, financial analysts, and shop workers to identify viable improvements to current practices. This article presents two case studies of efforts to enhance productivity and ESOH in corrosion control facilities at Randolph Air Force Base (AFB), Texas, and Robins AFB, Ga. At each site, activity based cost models were created to increase the visibility of ESOH-related costs and target improvement opportunities. Analysis of the strip-and-paint processes for the T-38 aircraft at Randolph and the F-15 radome and C-141 aft cowl at Robins revealed that a large proportion of operating costs were tied to ESOH requirements and practices (22 and 39%, respectively). In each case ESOH professionals teamed with shop personnel to identify potential improvements in personal protective equipment use, waste disposal, tool selection, and work methods. This approach yielded alternatives projected to reduce total shop costs by 5 to 7%. This case study demonstrates how workplaces can identify cost-saving and efficiency-enhancing practices by partnering with ESOH professionals in planning and decision-making activities. PMID- 14521431 TI - Development of a flow-injection fluorescence method for estimation of total polycyclic aromatic compounds in asphalt fumes. AB - Traditionally, measurements of specific polycyclic aromatic compounds (PACs) have been attempted as an estimate of asphalt fume exposure. However, asphalt fumes contain numerous alkyl substituted PACs, including PACs containing heteroatoms of nitrogen, oxygen, and sulfur. Many of these compounds coelute precluding the resolution of the individual compounds resulting in ambiguous data. Moreover, many researchers believe that some observed health hazards are associated with PACs overall and not just a few select PACs. Therefore, NIOSH method 5800 was developed to evaluate total PACs as a chemical class in asphalt fumes. Asphalt fume samples were collected on a poly(tetrafluoroethylene) filter backed by an XAD-2 sorbent tube. The samples were extracted with hexane; then, a cyano-solid phase-extraction column was used to remove the polar compounds while the aliphatic and aromatic compounds were eluted with hexane. An equal volume of dimethyl sulfoxide (DMSO) was added to the hexane extract, causing the aromatic compounds to partition into the DMSO, thus isolating the PACs. The PACs were then analyzed for fluorescence using a flow-injection method with two fluorescence detectors. Wavelength settings for the first detector (254-nm excitation, 370-nm emission) emphasized the 2- to 4-ring PACs that may cause eye and respiratory tract irritation. Wavelength settings of the second detector (254-nm excitation, 400-nm emission) emphasized the 4- and higher-ring PACs that are often mutagenic and possibly carcinogenic. PMID- 14521430 TI - An extended equation for rate coefficients for adsorption of organic vapors and gases on activated carbons in air-purifying respirator cartridges. AB - Organic vapor adsorption rates in air-purifying respirator cartridges (and other packed beds of activated carbon granules) need to be known for estimating service lives. The correlation of Lodewyckx and Vansant [AIHAJ 61:501-505 (2000)] for mass transfer coefficients for organic vapor adsorption onto activated carbon was tested with additional data from three sources. It was then extended to better describe all the data, including that for gases. The additional parameter that accomplished this was the square root of molar equilibrium capacity of the vapor or gas on the carbon. This change, along with skew corrections when appropriate, resulted in better correlations with all experimental rate coefficients. PMID- 14521432 TI - Confidence intervals for the mean of sound exposure levels. AB - This text transposes standard statistical estimators of the mean and its confidence intervals to the field of occupational noise exposure, assuming that the samples are independent and lognormally distributed. The hypothesis of lognormality is applied to the values of A-weighted sound exposure as defined in ISO 1999 and expressed in Pa(2).h, which is equivalent to the currently accepted hypothesis of normality applied to the values of equivalent continuous A-weighted sound pressure level L(Aeq,T) expressed in dB(A). By expressing the initial data and results in dB(A), the text provides an unbiased estimator of the mean sound exposure level and tables of confidence intervals according to the sample size n and the standard deviation S(L) of the L(Aeq,T) measured values. The values of the following confidence intervals are given: exact one-sided upper and lower 95 and 97.5% confidence intervals and exact two-sided symmetrical 95% confidence interval. These results are compared with the approximate two-sided symmetrical 95% confidence interval proposed in standard, ISO 9612. This comparison demonstrates that the use of the approximate confidence interval can markedly underestimate the upper limit of the confidence interval when n is small and if S(L) is above 3 dB(A). PMID- 14521433 TI - Maximum handgrip force in relation to upper limb posture--a meta-analysis. AB - It is essential to establish normative values of handgrip force in relation to factors influencing the force. This study developed a predictive equation that expresses the maximum force of the handgrip in relation to upper limb posture and gender. To create the equation, data from published studies on experimental results of maximum handgrip force in different upper limb postures were used. Selected were only those studies that describe upper limb posture during experiments clearly enough so that it could be transferred according to the Seven Degrees of Freedom Model, which unambiguously defines upper limb posture with values of seven angles. A predictive equation for male maximum handgrip force in relation to angles of wrist flexion/extension, wrist adduction/abduction, forearm pronation/supination, elbow flexion, shoulder flexion/extension, shoulder horizontal adduction/abduction, and arm medial/lateral rotation along the long axis was developed. Also developed was a mathematical formula that expresses maximum handgrip force for men in relation to maximum handgrip force for women. The equation is general and can be used for calculating norm values. It can also be applied to a specific population by performing an experimental study for one upper limb posture and assessing maximum force on the basis of the predictive equation for others. PMID- 14521434 TI - Methodologies for determining capture efficiencies in surface treatment tanks. AB - Methodologies are proposed for determining capture efficiencies in the ventilation systems of surface treatment tanks, using test-scale equipment. The equipment, which incorporates a lateral and push-pull ventilation system, can measure and control the variables of interest because it incorporates a tracer gas generator (sulfur hexafluoride, the concentration of which is measured by infrared spectrometer). The experimental methodologies described determine total efficiency (when the tracer is emitted uniformly from the whole surface of the tank) and the so-called transversal linear efficiency (when the tracer is emitted linearly through a perforated tube situated over the tank, parallel to the exhaust hood face). The analytical and graphical relationships that can be are established between the two efficiencies make it possible to detect where the emissions not captured by the ventilation system are produced (i.e., losses to the outside). At the same time, such losses can be quantified. Several experiments, results of which are analyzed by the methods described, are included. PMID- 14521435 TI - An occupational exposure assessment of a perfluorooctanesulfonyl fluoride production site: biomonitoring. AB - This investigation randomly sampled a fluorochemical manufacturing employee population to determine the distribution of serum fluorochemical levels according to employees' jobs and work areas. Previous analyses of medical surveillance data have not shown significant associations between fluorochemical production employees' clinical chemistry and hematology tests and their serum PFOS and perfluorooctanoate (PFOA, C(7)F(15)COO(-)) concentrations, but may have been subject to nonparticipation bias. A random sample of the on-site film plant employee population, where fluorochemicals are not produced, determined their serum concentrations also. Of the 232 employees randomly selected for serum sampling, 186 (80%) employees participated (n=126 chemical plant; n=60 film plant). Sera samples were extracted using an ion-pairing extraction procedure and were quantitatively analyzed for seven fluorochemicals using high-pressure liquid chromatography electrospray tandem mass spectrometry methods. Geometric means (in parts per million) and 95% confidence intervals (in parentheses) of the random sample of 126 chemical plant employees were: PFOS 0.941 (0.787-1.126); PFOA 0.899 (0.722-1.120); perfluorohexanesulfonate 0.180 (0.145-0.223); N-ethyl perfluorooctanesulfonamidoacetate 0.008 (0.006-0.011); N-methyl perfluorooctanesulfonamidoacetate 0.081 (0.067-0.098); perfluorooctanesulfonamide 0.013 (0.009-0.018); and perfluorooctanesulfonamidoacetate 0.022 (0.018-0.029). These geometric means were approximately one order of magnitude higher than those observed for the film plant employees. PMID- 14521436 TI - Comparison of a passive aerosol sampler to size-selective pump samplers in indoor environments. AB - The objective of this work was to investigate the ability of the Wagner-Leith passive aerosol sampler to measure indoor exposures over periods of 24 hours to 2 weeks. An automated analysis technique was developed so that lower aerosol concentrations could be sampled over shorter time periods. A test of the new analytical method against a manual method showed good agreement. The passive sampler was tested alongside three pump-operated, size-selective samplers in indoor environments. Generally, good correlation with the active samplers was observed. Correlation with a personal impactor with uncoated substrates was not statistically significant, but the cyclone, MS&T impactor, and overall correlations had R(2) values of 0.73-0.88. Combining these data with a previous study produced an R(2) of 0.96 between passive and active results. Large discrepancies (up to 147%) between passive and personal impactor results were observed and were attributed to particle bounce in the impactor, passive sampler imprecision due to few collected fine particles, and problems with detection of organic particles in the passive sampler. The Wagner-Leith sampler has now been tested over five orders of magnitude in mass concentration, in which it has proved useful for obtaining aerosol size distributions, mass fractions, qualitative elemental analysis, and morphology of individual particles. The sampler has several limitations, including increased sensitivity to contamination when fewer particles are collected, uncertainties in sampling semi-volatile particles, and the need for some expertise and expense to analyze the passive samples. PMID- 14521437 TI - Application of statistical models to estimate the correlation between urinary benzene as biological indicator of exposure and air concentrations determined by personal monitoring. AB - This study evaluated the correlation between benzene in urine and in workplace air at low airborne benzene levels (below 1 ppm). Eleven workers were monitored over a period of 1-4 days at a petrochemical plant in Italy; samples of end-of shift urine and workplace air were analyzed for benzene. A significant correlation, with a coefficient of determination R(2)=0.63, was found between urine and airborne benzene, confirming the results of previous studies. Two different statistical models were utilized to estimate urine benzene values of 9 16 microg/L corresponding to the American Conference of Governmental Industrial Hygienists' threshold limit value (TLV) of 0.5 ppm in workplace air. Not withstanding the variability inherent to biological monitoring, the results suggest application of biomonitoring as a trigger for identification of lower exposure level below, but approaching the TLV. Additionally, the proposed benzene biomonitoring may be useful in evaluating PPE effectiveness and use characteristics as well as dermal contribution to total exposure. PMID- 14521438 TI - Pathologic quiz case: a 72-year-old man with a well-circumscribed breast mass. Myofibroblastoma. PMID- 14521439 TI - Pathologic quiz case: chest wall mass in a 74-year-old man. Extraskeletal myxoid chondrosarcoma. PMID- 14521440 TI - Pathologic quiz case: a pelvic mass in a 20-year-old man. Diffuse large B-cell lymphoma with Homer-Wright-type rosette formation. PMID- 14521441 TI - Ossifying adult xanthogranuloma. AB - Although a xanthogranuloma is a relatively common benign cutaneous condition and ossification has been observed within many cutaneous lesions, the association between ossification and xanthogranuloma has not, to our knowledge, been reported previously. We believe we describe for the first time the case of a xanthogranuloma with marked osseous metaplasia on the trunk of a 41-year-old woman. Microscopically, the lesion showed typical features of a xanthogranuloma, with the exceptional feature of exuberant bone formation. The presence of bone within this lesion is likely secondary to a metaplastic process. PMID- 14521442 TI - Sarcomatoid carcinoma of the gallbladder with a rhabdoid tumor component. AB - We report the case of a sarcomatoid carcinoma with a rhabdoid tumor component originating in the gallbladder, along with immunohistochemical and electron microscopic findings. A 61-year-old woman presented with a 5-month history of right upper quadrant pain. Ultrasonography and a computed tomographic scan indicated gallbladder cancer. She underwent a cholecystectomy and a common bile duct resection. A firm mass (4.5 cm in greatest dimension) was present in the neck portion of the gallbladder. The mass was firm, solid, yellowish gray, and granular with areas of necrosis. Microscopically, the tumor was a biphasic sarcomatoid carcinoma and consisted of diffusely arranged pleomorphic cells, focally showing rhabdoid features and neoplastic glands with focal mucin production. Heterologous components such as osteoid, chondroid, and rhabdomyoblastic elements were not identified. By immunohistochemical staining, we demonstrated that the rhabdoid cells coexpressed cytokeratin and vimentin. On electron microscopic examination, the rhabdoid tumor cells showed cytoplasmic whorls of intermediate filaments in the cytoplasm and eccentric nuclei. Two months postoperatively, the follow-up computed tomographic scan showed multiple intrahepatic metastases and omental seedings. PMID- 14521443 TI - Hepatic adenomatosis in glycogen storage disease type Ia: report of a case with unusual histology. AB - Hepatic adenomatosis is a well-known complication of glycogen storage disease type Ia (von Gierke disease). Although most of these tumors have an appearance similar to sporadic hepatocellular adenomas, unusual histologic features have been reported, including Mallory hyaline, varying degrees of fibrosis, and aggregates of neutrophils. We report the fourth case of Mallory hyaline in the adenomas of glycogen storage disease type Ia in a 28-year-old woman undergoing segmental hepatectomy for enlarging liver nodules. Other prominent findings included steatohepatitis and nonspecific granulomatous inflammation--2 findings that are commonly seen in sporadic adenomas but not, to our knowledge, previously reported in glycogen storage disease type Ia. PMID- 14521444 TI - Synovial sarcoma with rhabdoid features. PMID- 14521445 TI - Howell-Jolly body-like inclusions in neutrophils. PMID- 14521446 TI - Pathologic quiz case: a woman with right facial nerve paralysis. Endolymphatic sac tumor associated with von Hippel-Lindau disease. PMID- 14521447 TI - Pathologic quiz case: a 49-year-old woman with a lower outer quadrant left breast mass. Matrix-producing metaplastic carcinoma of the breast. PMID- 14521448 TI - Pathologic quiz case: an 81-year-old woman with compression fracture and renal failure. Multiple myeloma with a monoclonal serum IgD lambda immunoglobulin. PMID- 14521449 TI - Pathologic quiz case: a 72-year-old man with fatigue and proteinuria. Angiotropic (intravascular) large B-cell lymphoma. PMID- 14521450 TI - Carcinoembryonic antigen elevation due to bowel sequestration with mucocele formation following colonic resection. AB - Carcinoembryonic antigen (CEA) is recommended as a serologic marker to monitor colorectal carcinoma recurrence. Elevations of CEA due to causes other than carcinoma exist and may lead to a misdiagnosis of recurrent carcinoma. We report a case of bowel sequestration with mucocele formation at the site of previous colo-colic anastomosis causing a mild elevation in CEA. The patient exhibited increasing CEA levels 6 years after resection of a sigmoid colon carcinoma with end-to-end anastomosis. Subsequently, computed tomographic and positron emission tomographic scans documented the presence of a cystic mass showing increased uptake at the anastomotic site. At exploratory laparotomy a mass lesion with mucus-filled protrusions was resected. Pathologic examination documented the presence of sequestration of a segment of the bowel wall with a mucocele and no overlying defect at the mucosal anastomotic site by demonstrating the presence of all bowel layers. After resection of the lesion, the CEA level normalized. PMID- 14521451 TI - Colonic carcinoid metastatic to the breast. AB - Metastatic tumors of the breast are uncommon. Breast metastases from nonmammary malignant neoplasms are rare, accounting for approximately 2% of all breast tumors. We report the case of an ileal carcinoid tumor metastatic to the breast 10 years after the initial diagnosis. A 53-year-old woman presented to our clinic with a palpable breast lump. The mammogram was nonspecific. A lumpectomy was performed that, on frozen section, showed a neoplastic lesion. Permanent sections showed that the tumor was composed of sheets of small uniform cells divided into lobules by delicate vascular septa. Immunohistochemical analysis showed that the lesional cells were strongly positive to the neuroendocrine marker panel of antibodies: chromogranin A, neuron-specific enolase, synaptophysin, serotonin, and low-molecular-weight keratin. The lesional cells were negative to cytokeratins 7 and 20, estrogen and progesterone receptors, carcinoembryonic antigen, and c-Erb-B2 antibodies. The presence of pleomorphic neurosecretory-type granules within the cytoplasm of the tumor cells by ultrastructural analysis strongly suggested a metastatic lesion from a midgut carcinoid. A detailed review of the patient's medical records confirmed a right hemicolectomy for an ileal carcinoid with lymph node and omental metastases that had been performed elsewhere 10 years earlier. A detailed pathologic analysis of this lesion by light microscopy, along with histochemical, immunohistochemical, and ultrastructural analyses, aided in confirming the metastatic nature of the current breast lesion. PMID- 14521452 TI - Combined serous microcystic adenoma and well-differentiated endocrine pancreatic neoplasm: a case report and review of the literature. AB - We report a case of combined microcystic adenoma and pancreatic endocrine neoplasm of the pancreas in a 53-year-old male patient. The pancreatic tumor was an incidental computed tomography scan finding and was not accompanied by gastrointestinal symptoms. The tumor was located in the head of the pancreas and was composed of numerous small cysts lined by uniform clear cells with a centrally located solid endocrine component. Four cases of similar neoplasm have recently been reported, exclusively in women. Literature review and case analysis indicate that combined microcystic adenoma and pancreatic endocrine neoplasm is characterized by the presence of pancreatic endocrine neoplasm within microcystic adenoma in the head of the pancreas, affects women more often than men, and presents at a younger age when compared to microcystic adenoma. PMID- 14521453 TI - Severe hemolysis due to passenger lymphocyte syndrome after hematopoietic stem cell transplantation from an HLA-matched related donor. AB - A 61-year-old white man (group A, Rh-positive) was allotransplanted for acute myelogenous leukemia from his HLA-matched related sister (group O, Rh-positive) in 2 separate infusions. Three days after the second graft infusion, the patient's front blood type converted to O Rh-positive, with a negative direct antiglobulin test and elevated anti-A1 titer. Severe hemolysis developed, and the patient expired 14 days posttransplantation. PMID- 14521454 TI - Congenital anerythremic erythroleukemia presenting as hepatic failure. AB - We report an atypical case of congenital erythroleukemia in a child born with hepatosplenomegaly and abnormal liver tests. The initial peripheral blood cell count showed anemia and hyperleukocytosis with erythroblastosis that disappeared 1 week later. During the next 5 weeks, no blasts were found in the blood, and less than 5% were found on 2 successive bone marrow aspirates. The infant died of hepatic failure. The suspected diagnosis on a premortem liver biopsy was confirmed by an autopsy that showed a blastic infiltration in many organs. These cells expressed only erythroid markers glycophorin A and C. Rearrangement of the myeloid lymphoid leukemia gene was not found by fluorescence in situ hybridization. The main differential diagnoses include metabolic diseases, Langerhans histiocytosis, Pepper syndrome, transient myeloproliferative disorder, and leukemoid reactions. Although some of these can be excluded by the pathologist, others require a multidisciplinary confrontation: clinical, biologic, genetic, and pathologic examinations. PMID- 14521455 TI - Fatal dural sinus thrombosis associated with heterozygous factor V Leiden and a short activated partial thromboplastin time. AB - Inherited thrombophilia is a risk factor for dural sinus thrombosis (DST). To our knowledge, this is the first description with autopsy findings of a patient with DST associated with heterozygous factor V Leiden and a short activated partial thromboplastin time (aPTT). A 51-year-old woman presented with a 3-day history of headache, nausea, right-sided weakness, and focal motor seizure; she died 3 days after admission. At autopsy, a gross examination showed hemorrhage of bilateral parietal lobes and left primary motor cortex, uncal and tonsillar herniation, and pulmonary embolus of the right upper lobe. A microscopic examination of the brain showed an organizing thrombus in the superior sagittal sinus, diffuse cerebral edema, and extensive venous congestion. Laboratory studies showed heterozygous factor V Leiden by polymerase chain reaction and a very short aPTT of 17 seconds (reference range, 22-30 seconds). The combination of a heterozygous factor V Leiden mutation and a short aPTT may have contributed to the fatal DST in this patient. PMID- 14521456 TI - Quality assurance in human molecular genetics testing: status and recommendations. AB - OBJECTIVE: The role of genetic testing has expanded with rapidly developing technology and completion of the International Human Genome Project. Development of universally acceptable quality control methods and quality assurance standards trails technology. The principle that high-quality genetic testing is important for public health motivated the Centers for Disease Control and Prevention to formulate ways for improving quality assurance of human molecular genetics testing. PARTICIPANTS: Twenty-eight panelists were chosen based on expertise in molecular genetics testing and knowledge of quality assurance practices. Representatives of professional organizations, industries, and federal agencies participated in one or more of 3 panel meetings. Consensus recommendations were developed by the 15 panelists in the third meeting. EVIDENCE: Evidence was derived from experts' opinion during 3 panel meetings. Data compiled through laboratory visits and literature review were used as reference information. Need for this project was derived from the Final Report of the Task Force on Genetic Testing, produced by the National Institutes of Health and the Department of Energy in 1997, and the Summary Report of the Subcommittee Meeting on Genetics of the Clinical Laboratory Improvement Act Advisory Committee in 1997. CONSENSUS PROCESS: Research and development needs were identified using a participatory visioning approach. A modified nominal group process was used to reach consensus. CONCLUSIONS: Five core consensus recommendations were made: research for developing positive samples for quality assurance purposes, performance evaluation programs supplementing those in existence, establishment and support of laboratory-oriented consortia, establishment of a laboratory-focused database, and support of molecular genetics training programs. PMID- 14521457 TI - Prevalence and role of methylenetetrahydrofolate reductase 677 C-->T and 1298 A- >C polymorphisms in coronary artery disease in Arabs. AB - CONTEXT: Previous studies reported an association of 677 C-->T and 1298 A-->C methylenetetrahydrofolate reductase (MTHFR) variants with coronary artery disease (CAD). No previous studies concerning the prevalence of these 2 MTHFR variants or their possible association with CAD in Arabs are currently available in the literature. OBJECTIVE: To determine the prevalence of MTHFR variants and their potential relevance to CAD among Arabs. DESIGN: We used polymerase chain reaction and restriction enzyme digestion to determine the prevalence of these 2 MTHFR polymorphisms in 625 healthy blood donors (BDs) and 545 angiographically confirmed CAD patients of Arab origin. RESULTS: For the 677 C-->T variant within the CAD group, 64.2% were homozygous wild-type C/C, 32.1% were heterozygous C/T, and 3.7% were homozygous T/T genotype. Within the BD group tested for the 677 C- >T variant, 72.2% were homozygous wild-type C/C, 25.8% were heterozygous C/T, and 2% were homozygous T/T genotype. Within the CAD group tested for the 1298 A-->C variant (n = 540), 45.7% were homozygous wild-type A/A, 46.9% were heterozygous A/C, and 7.4% were homozygous C/C genotype. Within the BD group tested for the 1298 A-->C variant (n = 625), 39.4% were homozygous wild-type A/A, 51.5% were heterozygous A/C, and 9.1% were homozygous C/C genotype. The distribution and allele frequency of these 2 MTHFR variants followed the Hardy-Weinberg equilibrium and were similar in the CAD and BD study groups. The prevalence of the 677 C-->T and 1298 A-->C compound heterozygosity was 9.6% for the BD group and 12.3% for the CAD group. CONCLUSION: The 2 MTHFR variants tested in this study, individually or compound, are not associated with CAD. Therefore, neither of these 2 variants can be considered an independent risk factor or a predictor for CAD in this population. PMID- 14521458 TI - Kikuchi-Fujimoto disease in Nepal: a study of 6 cases. AB - CONTEXT: Kikuchi-Fujimoto disease is a benign disorder, predominantly affecting young women with a predilection for cervical lymphadenopathy. Although the disease has been recognized worldwide, to our knowledge no cases have been reported previously from Nepal. OBJECTIVES: To determine the prevalence of Kikuchi-Fujimoto disease in Nepal and to analyze clinicopathologic features. METHODS: We reviewed 6 cases of Kikuchi-Fujimoto disease recorded at 3 different hospitals in Nepal during a period from June 1998 to June 2002. Clinical data and histopathology are presented. RESULTS: This study included 5 females and 1 boy, aged 13 to 32 years. These patients presented with prolonged fever and lymphadenopathy. The 5 female patients had cervical lymphadenopathy, and the boy had axillary lymphadenopathy. Complete blood counts revealed raised erythrocyte sedimentation rates in all patients and anemia in 2 patients. The size of excised lymph nodes (in greatest dimension) ranged from 1.5 to 5 cm. Typical histologic features were seen, namely, architectural effacement due to presence of pale nodular lymphohistiocytic foci with karyorrhectic debris, coagulation necrosis, eosinophilic debris, and absence of granulocytic infiltration. In a follow-up of the cases, disease recurrence was not found. CONCLUSION: Our study emphasizes that Kikuchi-Fujimoto disease should be considered as one of the differential diagnoses in patients with prolonged fever and cervical lymphadenopathy and that it should be differentiated from tuberculous lymphadenitis in regions where tuberculosis is prevalent. PMID- 14521459 TI - Is B-lineage acute lymphoblastic leukemia with a mature phenotype and l1 morphology a precursor B-lymphoblastic leukemia/lymphoma or Burkitt leukemia/lymphoma? AB - CONTEXT: B-lineage acute lymphoblastic leukemia (ALL) with a mature phenotype and L1 morphology is a rare condition that may pose a diagnostic and management challenge. OBJECTIVE: To report our experience with 2 such unusual cases of pediatric B-lineage ALL. DESIGN: Morphologic, immunophenotypic, and cytogenetic features of the leukemic blast cells were reviewed in conjunction with clinical and other laboratory findings. RESULTS: The leukemic blast cells in both cases were small to medium with scant basophilic cytoplasm and several small inconspicuous nucleoli, characteristic of L1 lymphoblasts. Immunophenotypically, they were positive for CD19, CD22, and low-density CD20, with expression of surface immunoglobulin lambda light chain. They were negative for immature (CD34 and terminal deoxynucleotidyl transferase), myeloid, and T-cell-associated markers. Conventional cytogenetic and fluorescent in situ hybridization studies failed to demonstrate chromosomal translocations involving the c-myc gene. Both patients were treated with Children's Cancer Group ALL protocols and had good responses. CONCLUSIONS: B-lineage ALL with a mature phenotype, L1 morphology, and absent chromosomal translocations involving the c-myc gene is best classified and managed as precursor B-lymphoblastic leukemia/lymphoma instead of Burkitt leukemia/lymphoma. PMID- 14521460 TI - Latent membrane protein 1, tumor necrosis factor receptor-associated factor (TRAF) 1, TRAF-2, TRAF-3, and nuclear factor kappa B expression in posttransplantation lymphoproliferative disorders. AB - CONTEXT: Most posttransplantation lymphoproliferative disorders (PTLDs) are associated with Epstein-Barr virus (EBV) infection. The EBV latent membrane protein 1 (LMP-1) is important in the transformation of B lymphocytes through its interaction with intracellular tumor necrosis factor receptor-associated factors (TRAFs) that, in turn, can activate transcription factors such as nuclear factor kappa B (NFkappaB) and Jun-N-kinase. Of the 6 members of the TRAF family, TRAF-1, TRAF-2, and TRAF-3 are most commonly associated with LMP-1. Recently, it has been suggested that LMP-1-induced TRAF activation is important in the pathogenesis of PTLDs. OBJECTIVE: To characterize the expression patterns of these proteins in PTLDs, we studied a series of well-characterized cases for expression of LMP-1, TRAF-1, TRAF-2, TRAF-3, and NFkappaB by immunohistochemical analysis. METHODS: A total of 27 specimens from 25 patients were analyzed for LMP-1, TRAF-1, TRAF-2, TRAF-3, and NFkappaB (active form) by immunohistochemical analysis. Expression of EBV-encoded RNA (EBER) was evaluated by in situ hybridization. Correlation between the expression of the different markers was performed using the Mantel Haenszel chi(2) test. Cox proportional hazards analysis and Kaplan-Meier analysis with log-rank testing were used to analyze antigen expression and clinical outcome. RESULTS: Ninety-six percent of PTLDs expressed NFkappaB, 74% to 84% expressed TRAFs, 78% expressed EBER, and 77% expressed LMP-1. TRAF-1, TRAF-2, and TRAF-3 expression did not correlate with either EBER or LMP-1 expression. TRAF-2, but not TRAF-1 or TRAF-3, expression correlated with NFkappaB expression (P =.02). CONCLUSIONS: These results suggest that TRAF molecules and active NFkappaB are expressed in PTLDs regardless of EBV positivity. Given the association of TRAF-2 and active NFkappaB expression, TRAF-2 may play an important role in regulating this transcription factor in PTLD. PMID- 14521461 TI - Mammaglobin expression in lymph nodes is an important marker of metastatic breast carcinoma. AB - CONTEXT: Organ specificity is a desirable property of a tumor marker, especially in metastatic adenocarcinomas of unknown primary origin. Mammaglobin, a mammary specific member of the uteroglobin family, is known to be overexpressed in human breast cancer. OBJECTIVE: We investigated mammaglobin A expression in metastatic carcinomas of lymph nodes from the breast and various other organs and its usefulness in identifying metastatic carcinoma of the breast. For comparative purposes, we also investigated BRST-1 and BRST-2 expression. DESIGN: We produced recombinant mammaglobin and polyclonal antimammaglobin antibodies. Mammaglobin expression was analyzed by immunohistochemical staining using a tissue microarray and by reverse transcription-polymerase chain reaction in 210 carcinomas, including those of the breast (n = 70), lung (n = 30), stomach (n = 30), colorectum (n = 25), hepatobiliary tract (n = 20), urinary tract (n = 10), thyroid gland (n = 10), ovary and endometrium (n = 10), and salivary gland (n = 5). RESULTS: Mammaglobin expression was observed in 59 cases (84.3%) of breast cancer and in 21 cases (15.0%) of nonbreast cancer. The BRST-1 and BRST-2 expression rates were 75.7% and 44.3% in breast cancer and 26.4% and 2.1% in nonbreast cancer, respectively. Mammaglobin is superior to BRST-1 for both specificity and sensitivity and is superior to BRST-2 for sensitivity. CONCLUSION: Our data suggest that mammaglobin is one of the first relatively mammary-specific and mammary-sensitive markers. Mammaglobin and BRST-2 appear to represent useful markers for breast cancer and should be used as a component of panels evaluating tumors of unknown primary sites. PMID- 14521462 TI - Comparison of Epstein-Barr virus presence in Hodgkin lymphoma in pediatric versus adult Argentine patients. AB - CONTEXT: In developed countries and high socioeconomic groups, Hodgkin lymphoma has an initial peak in young adulthood, whereas in undeveloped countries and low socioeconomic groups, it shows an early childhood peak. In developing countries, 90% of children are infected with the Epstein-Barr virus (EBV) by the age of 6 years, but in developed countries, only 30% to 40% are seropositive by that age. Early childhood EBV infection in 75% of Argentine patients was demonstrated. OBJECTIVE: To explore the epidemiology of Hodgkin lymphoma and its relationship with EBV in Argentine patients. METHODS: The presence of EBV was assessed by Epstein-Barr encoded RNA in situ hybridization and latent membrane protein 1 immunohistochemistry. We studied 92 pediatric and 42 adult Hodgkin lymphoma cases from a public center as well as 39 adult cases from a private center. RESULTS: The mixed cellularity Hodgkin lymphoma had a prevalence of 52% in the pediatric group, while similar frequencies of both nodular sclerosis Hodgkin lymphoma (47%) and mixed cellularity Hodgkin lymphoma (44%) were observed in adults. As for Epstein-Barr encoded RNAs, 55% of the pediatric cases and 31% of the adult cases were positive. Among adult EBV+ cases, 38% were from the public hospital, and 23% were from the private center. EBV was present in 77% of the pediatric mixed cellularity Hodgkin lymphoma cases when compared with the other histologic subtypes of Hodgkin lymphoma. EBV was mainly detected in mixed cellularity cases (39% in the adult group). CONCLUSION: Our findings strengthen the argument that EBV is involved in the pathogenesis of Hodgkin lymphoma in most children younger than 10 years. Our findings of EBV prevalence, along with both childhood and second-decade peaks as well as comparable frequencies for Hodgkin lymphomas of mixed cellularity and nodular sclerosis, distinguish our population from others in developing countries. PMID- 14521463 TI - Virtual microscopy as a tool for proficiency testing in cytopathology: a model using multiple digital images of Papanicolaou tests. AB - BACKGROUND: Modern digital cameras can acquire images from cytologic slides at sufficient resolution to allow for digital enlargement and scrolling on a video monitor, allowing for the simulation of microscopy using a computer. OBJECTIVE: The purpose of this study was to develop a tool for proficiency testing in cytopathology using multiple digital images of Papanicolaou tests. METHODS: Nine images were photographed from each of 10 Papanicolaou tests at x100 optical magnification, 3400 x 2300-pixel resolution, using a light microscope and a digital camera. All images from each case were tiled in a single canvas with Photoshop 4.0 software. Two cytopathologists and 3 cytotechnologists interpreted these "virtual slides" using a computer and graded diagnostic codes (PAP program, College of American Pathologists). Subjects were retested a year later using the glass slides from the same cases and routine microscopy. Both test results, by diagnostic code, were compared with the McNemar test of symmetry. RESULTS: The 5 test subjects provided 42 and 50 correct diagnostic codes by "virtual microscopy" and light microscopy, respectively. No significant asymmetry in results obtained by virtual microscopy and light microscopy was encountered with the McNemar test of symmetry. All test answers were correctly classified by selection series, using both virtual microscopy and light microscopy, and the responses would have been graded as 100% by current PAP program scoring guidelines. This suggests that virtual microscopy could be used for proficiency testing purposes. CONCLUSIONS: A simple virtual microscopy method designed to challenge participants to locate and diagnose cells of interest was effective for the administration of standardized proficiency tests. Virtual microscopy methods that rely on single-plane images to locate and diagnose cells of interest could provide effective proficiency testing tools prior to the development of more computationally intensive systems that represent an entire Papanicolaou test at multiple focal planes. PMID- 14521464 TI - Schnabel cavernous degeneration: a vascular change of the aging eye. AB - CONTEXT: Schnabel cavernous degeneration is a histologic finding originally attributed to glaucoma; however, its cause and significance have been controversial. OBJECTIVE: To determine the basic cause of the formation of cavernous spaces in the proximal optic nerve and its clinical significance. METHODS: A retrospective analysis of 4500 autopsy eyes processed for histologic evaluation between 1967 and 1991. RESULTS: Ninety-three (2.1% of eyes examined) cases of Schnabel cavernous optic atrophy were identified. The majority of the eyes were from women (81%). The mean age of the entire group was 88 years (reference range, 54-103 years). Severe vascular anomalies were present in 75% of the individuals. Cavernous degeneration was unilateral in 82% of the cases. Loss of ganglion cells and nerve fiber layer consistent with glaucoma was found in 23.7% of the individuals. Clinical information was available for 15 individuals (16%). Half of them were thought to have some clinical optic nerve damage; in the remainder, no specific optic disc abnormalities were noted. Histologic findings of arteriolosclerosis in the optic nerve circulation were common. The prevalence of glaucoma was low. CONCLUSIONS: Schnabel cavernous optic atrophy appears to be a unilateral condition of elderly women with systemic vascular disease and few characteristic ocular features. Our data indicate that a chronic vascular occlusive disease of the proximal optic nerve is more involved in cavernous atrophic pathogenesis than is a sustained increase of intraocular pressure. PMID- 14521465 TI - Updated protocol for the examination of specimens from patients with carcinoma of the lung. PMID- 14521466 TI - Protocol for the examination of specimens from patients with thymic epithelial tumors located in any area of the mediastinum. PMID- 14521467 TI - Protocol for the examination of specimens from patients (children and young adults) with rhabdomyosarcoma. PMID- 14521468 TI - Protocol for the examination of specimens from patients with Wilms tumor (nephroblastoma) or other renal tumors of childhood. PMID- 14521469 TI - Updated protocol for the examination of specimens from patients with carcinoma of the urinary bladder, ureter, and renal pelvis. PMID- 14521470 TI - Protocol for the examination of specimens from patients with melanoma of the skin. PMID- 14521472 TI - Problems with the National Cholesterol Education Program recommendations for cholesterol analytical performance. PMID- 14521473 TI - Are College of American pathologists accreditation standards relevant outside North America? PMID- 14521474 TI - Spinocerebellar degeneration. AB - The spinocerebellar degenerations/ataxias (SCAs) are a diverse group of rare, slowly progressive, neurological diseases, often inherited but of incompletely understood pathophysiology, which affect the cerebellum and its related pathways. They have few animal models and share no reliable biomarkers. They have, as yet, no universally validated rating scale for use in clinical trials. In the past 25 years, there have been, at most, 18 controlled (Class 1) trials for ataxia, which have focused on neurotransmitter mechanisms. There is currently only one National Institute of Neurological Disorders and Stroke-sponsored drug trial for ataxia (Phase I study of idebenone in Friedreich's ataxia). There are, as yet, no FDA approved drugs for SCA. Current treatment practices encompass rehabilitation interventions and off-label use of symptomatic medications [1,2]. PMID- 14521475 TI - Pharmacotherapy for personality disorders. AB - Although there are many methodological uncertainties in the treatment of personality disorders, which make it difficult to be confident about efficacy, there is growing evidence that at least in some personality disorders, drug treatment may be of value. Borderline personality disorder is a condition in which treatment evidence is the most promising, but it is also one of the most difficult disorders to disentangle the mental state from personality components. In summary, there is reasonable evidence that antidepressants, particularly serotonin re-uptake inhibitors and monoamine oxidase inhibitors, have beneficial effects independent of their antidepressive ones and albeit, less favourable, evidence that antipsychotic drugs and mood stabilisers may also be of value. None of this evidence is yet sufficient to point to any specific drug treatment indications. PMID- 14521476 TI - Pharmacotherapy of postpartum psychosis. AB - The term postpartum psychosis (PP) is not a discrete nosological entity but rather, refers to a group of heterogeneous disorders that share the common features of the presence of psychosis and onset of symptoms during the postpartum period. Although there remains a debate regarding its diagnostic status, PP is generally considered as belonging to the bipolar spectrum. The prognosis for the individual episodes is considered generally favourable but it is a potentially serious illness due to the increased risk of suicide and infanticide. Furthermore, puerperal and non-puerperal recurrences are quite common. There is paucity of controlled trials in the acute and prophylactic treatment of PP and the data are limited largely to the use of lithium and oestrogen. Strategies for the clinical management of PP should include early identification of women at risk; close monitoring of mood state during pregnancy, prompt recognition of impending psychosis and aggressive pharmacotherapy. In general, treatment for PP is essentially the same as for similar non-puerperal episodes. PMID- 14521477 TI - Towards the pharmacotherapy of eating disorders. AB - The purpose of this review is to discuss pharmacological options for the treatment of patients with eating disorders. Sequentially described are pharmacotherapy studies of anorexia nervosa (AN), bulimia nervosa (BN) and binge eating disorder (BED). The quantity of drug trials performed with AN patients has been very limited. While the majority of studies have failed to show medication efficacy for the acute treatment of AN, there is data which suggests that fluoxetine hydrochloride may play a role in preventing relapse during maintenance therapy. Atypical antipsychotics, most often olanzapine, have shown promise in a number of uncontrolled studies. BN has been most extensively studied, with the majority of pharmacological trials focusing on antidepressants. Fluoxetine, at a dose of 60 mg/day, is FDA-approved for the treatment of BN. Psychotherapy, particularly cognitive behavioural therapy (CBT) is of well-established utility in BN and data suggests that the combination of an antidepressant plus CBT is superior to either treatment alone. Recently, there has been interest in the 5 HT3 antagonist, ondansetron, and the anticonvulsant, topiramate. BED investigators have focused largely on antidepressants, which may reduce symptoms of depression and augment psychotherapy. While sibutramine and topiramate have both been associated with weight loss in controlled trials, the former appears to be fairly well-tolerated and the latter appears to be responsible for the emergence of significant cognitive and peripheral nervous system side effects in some patients. Further pharmacological research with eating disorder patients is needed, particularly in the areas of AN and BED. Also, pharmacological augmentation strategies for those not responding to primary therapies should be explored. PMID- 14521478 TI - Pharmacotherapy to limit weight gain caused by antipsychotic use. AB - Obesity is a major health problem, especially in patients with severe mental illnesses. Newer atypical antipsychotics are better tolerated and have fewer extrapyramidal side effects, however, they can cause significant weight gain. This poses a considerable added burden and increases the likelihood of developing medical illnesses such as diabetes and cardiovascular disease. Obesity in patients taking antipsychotic medication can usually be avoided, if behavioural modification and pharmacological measures are implemented from the outset. Greater awareness and a willingness to address the problem are needed along with more research into the development of weight gain and its prevention. This review examines these issues with a pharmacotherapy focus. PMID- 14521479 TI - Current trends in fibromyalgia research. AB - The development of standardised criteria for the diagnosis of fibromyalgia in 1990 has allowed careful study of this chronically painful syndrome. Epidemiological studies show increased symptoms and disability in patients with fibromyalgia, compared with other conditions associated with chronic, widespread pain. In addition, prevalence and severity of fibromyalgia symptoms are increased in women. Current studies have identified strong evidence for central sensitisation in fibromyalgia. Data from these studies may expand effective treatment options for fibromyalgia. PMID- 14521480 TI - Combination strategies for pain management. AB - At least two factors relating to pain management using oral analgesics suggest that combination strategies merit consideration: many pains arise from more than one physiological cause and current analgesics have adverse effect profiles that might be reduced by combination with another agent in smaller doses or with less frequent dosing. In addition to increased convenience, combinations sometimes also result in the unexpected benefit of synergy. But not all pains, clinical settings or combinations merit the extra expense or other potential negative features of fixed-ratio products. This review examines the multiple basic science, clinical and pharmacoeconomic issues relating to analgesic combinations and the methodologies available for assessing these issues. PMID- 14521481 TI - Central nervous system abnormalities in migraine. AB - Migraine is associated with structural and functional CNS changes, for example, ictal hyperalgesia and allodynia and interictal neural excitation. Structural abnormalities, most notably white matter changes, occur in greater prevalence in migraineurs (16 - 40%). Several studies have examined the neuropsychological correlates of migraine and/or white matter abnormalities. These studies suggest mild, interictal dysfunction in migraineurs. More research is needed to correlate migraine severity, frequency and/or treatment with neuropsychological testing. Additional studies should: identify interictal cognitive changes; clarify the contribution to long-term cognitive changes from migraine genotype, sequelae of repeated pain episodes or their treatment and the consequences of co-morbid vascular disease; and include cognitive measures as secondary end points in clinical trials. PMID- 14521482 TI - Cannabinoids reduce symptoms of Tourette's syndrome. AB - Currently, the treatment of Tourette's syndrome (TS) is unsatisfactory. Therefore, there is expanding interest in new therapeutical strategies. Anecdotal reports suggested that the use of cannabis might improve not only tics, but also behavioural problems in patients with TS. A single-dose, cross-over study in 12 patients, as well as a 6-week, randomised trial in 24 patients, demonstrated that Delta9-tetrahydrocannabinol (THC), the most psychoactive ingredient of cannabis, reduces tics in TS patients. No serious adverse effects occurred and no impairment on neuropsychological performance was observed. If well-established drugs either fail to improve tics or cause significant adverse effects, in adult patients, therapy with Delta9-THC should be tried. At present, it remains unclear whether herbal cannabis, different natural or synthetic cannabinoid CB1-receptor agonists or agents that interfere with the inactivation of endocannabinoids, may have the best adverse effect profile in TS. PMID- 14521483 TI - Current treatment options for restless legs syndrome. AB - Restless legs syndrome (RLS) is a common but often underdiagnosed neurological disorder characterised by an imperative desire to move the extremities associated with paraesthesias, motor restlessness, worsening of symptoms at rest in the evening or at night and, as a consequence, sleep disturbances particulary. Additionally, most patients with RLS have periodic limb movements during sleep and relaxed wakefulness. The aetiology of RLS remains unknown. Treatment of RLS is generally symptomatic, a causal therapy is possible only in the secondary forms. Dopaminergic agents including levodopa and dopamine agonists such as pergolide, pramipexole, cabergoline and ropinirole are regarded as the treatment of choice for idiopathic RLS, however, the development of augmentation of symptoms, especially under levodopa therapy, may be a major problem. Except in special circumstances, opioids and anticonvulsants such as gabapentin or benzodiazepines, are regarded as second-line treatment. In secondary RLS, the underlying illness should first be treated, although dopaminergic drugs may also be helpful. PMID- 14521484 TI - Pharmacological management of narcolepsy. AB - Narcolepsy is a disabling disorder characterised by excessive daytime sleepiness, cataplexy, hypnagogic hallucinations, sleep paralysis and disturbed nocturnal sleep. Traditionally, a wide variety of substances are used in the symptomatic pharmacological treatment of narcolepsy. Several generally more-tolerated substances have been added to the therapeutic repertoire after extensive testing in large patient populations during recent years. This review addresses the state of-the-art knowledge about the pharmacological treatment of narcolepsy along with the personal view of the authors. The recent discovery that narcolepsy is caused by deficient hypocretin (orexin) transmission opens a perspective on causal therapy. PMID- 14521485 TI - Rationale for current therapies in Parkinson's disease. AB - Parkinson's disease (PD) is a neurodegenerative disorder that affects an estimated 1 million people in the US and tens of millions worldwide. Medication therapy has made significant advances and improvements especially over the last 10 years. A number of new treatments and new strategies have emerged and the quality of life for the average sufferer has improved. This review will describe the rationale and strategies for current medical therapies in PD, with special emphasis on the use of antipsychotic agents. Levodopa remains the most efficacious medication for the management of PD. Long-term use of levodopa, however, is associated with the development of motor fluctuations including dyskinesia. Trials with dopamine agonists have demonstrated a delay in the onset of dyskinesia with the use of this therapy. There is also active, ongoing investigation to determine whether a neuroprotective effect may be present with agonist therapy. Anticholinergics have been successfully used to treat tremor as well as sialorrhoea and urinary urgency. Catechol-O-methyltransferase inhibitors increase 'on time', decrease 'off time,' and improve motor scores. Continuous stimulation of dopamine receptors may decrease the fluctations observed with pulsatile delivery of anti-Parkinsonian medications, but this will require more study. Monoamine oxidase-B inhibitors, specifically selegiline, may provide symptomatic improvement; the question as to whether a neuroprotective benefit is present remains unanswered. Amantadine has demonstrated both symptomatic benefit and dyskinesia benefit in some patients. Selective dopamine blockers such as clozaril and quetiapine, have been shown to be effective in the treatment of psychosis. This class of medications is particularly useful as an adjunctive to levodopa and dopamine agonists. Doses of dopaminergic drugs can be escalated to treat Parkinsonian symptoms, whereas selective dopamine blockers can be added to block psychosis. Old management strategies required a reduction in dopaminergic therapy and therefore worsened Parkinsonian symptoms. Even though there have been great advances in the medical options for symptomatic management of PD, there are still many unmet needs for this patient population. PMID- 14521486 TI - Pharmacotherapy of iron overload in thalassaemic patients. AB - The recommended treatment for thalassaemia major is regular blood transfusions, although these lead to the harmful accumulation of iron in the body. If untreated, iron overload is responsible for heart, liver and endocrine diseases. The only two iron chelating agents available for the treatment of iron overload are deferoxamine and deferiprone. The standard iron chelation therapy is based on the use of deferoxamine. Although this drug was introduced in the 1970s, it still remains the treatment of choice. Recently, another iron chelator, deferiprone, became available for clinical use in the European Community. Deferiprone is indicated as second-line treatment in patients with thalassaemia major, for whom deferoxamine therapy is contraindicated or in patients who present with serious toxicity to deferoxamine therapy. This paper examines this chelating agent and compares it with deferoxamine in order to ascertain the current and potential contribution of deferiprone to the treatment of thalassaemic patients. PMID- 14521487 TI - Assessing the cost-effectiveness of switching from a beta-blocker to latanoprost in the treatment of ocular hypertension. AB - Glaucoma is a pathological condition whose most important risk factor is increased intraocular pressure (IOP). The medical treatment of glaucoma essentially consists of compounds that are able to decrease the IOP. The compounds discussed in this review act in a different way, beta-blockers mainly inhibit the production of aqueous humor, whereas latanoprost decreases the resistance in the outflow channels. beta-Blockers are compounds with a well-known efficacy and safety profile and they are fairly inexpensive. Their systemic and local side effects are mainly cardiovascular and pulmonary adverse events, dry eye and keratopathy. Latanoprost, which has recently been introduced into the market, has been shown to be equally as effective, or better in lowering IOP in patients than timolol, although it is more expensive. Systemic reported side effects are anecdotal; local hyperaemia, keratopathy, hypertrichosis, increased pigmentation of eyelashes and iris, uveitis and cystoid macular oedema have been reported. A comparison of costs reveals that a 1-year therapy with timolol ophthalmic solution starts from 11.00 Euros and can reach 146.00 Euros for the most expensive preservative-free 1-day dispenser packages (approximately 13.5 times higher). For latanoprost once-daily administration, the cost for 1 years therapy is 98.55 Euros, approximately six times higher than generic or brand 0.5% timolol applied twice-daily. What are the factors influencing a change in therapy from beta-blockers to latanoprost? The only good reason is represented by a further deterioration in the visual field. This may occur, despite a significant reduction in IOP, because the reached IOP is not sufficient enough to avoid further deterioration because the patient's work or social activities do not allow a correct daily dosage of the compound (bad compliance); or as a result of treatment suspension, because of the development of systemic and/or local side effects. Changes in therapy must always be related to a failing control of the disease, as any therapeutic modification leading to an increase in the number of visits and additional examinations, consequently enhances the costs. PMID- 14521488 TI - The epidemiology of severe sepsis syndrome and its treatment with recombinant human activated protein C. AB - Severe sepsis syndrome has important consequences to healthcare systems as the incidence is increasing, there is significant attributed morbidity and mortality and there is a substantial cost for in-hospital and post-discharge care. Current treatment includes the use of antimicrobials, local source control and aggressive physiological support, usually in an intensive care unit setting. Drotrecogin alpha (activated) or recombinant human activated protein C (rhAPC) is the only biological agent approved for use in severe sepsis syndrome that has demonstrated efficacy in reducing 28-day all-cause mortality and new data suggests a trend towards longer term survival. However, given the evidence of a variable effect on survival rates in patient subgroups and its acquisition cost, controversy has arisen concerning its appropriate use. This review discusses the epidemiology of sepsis, preclinical and clinical evidence supporting the use of rhAPC use, controversies about the evidence of efficacy in severe sepsis syndrome and cost effectiveness data. PMID- 14521489 TI - Effectiveness of drug therapies to treat or prevent respiratory syncytial virus infection-related morbidity. AB - Respiratory syncytial virus (RSV) infection causes a huge burden to the health service, as it results in a large number of in-patient days each year and increases the risk of asthma in childhood. In the acute phase, therapy is supportive as bronchodilators and corticosteroids have resulted, at best, only in short-term benefits; promising treatments for ventilated patients, such as exogenous surfactant, require further testing. Passive immunoprophylaxis reduces hospital admission in high risk groups. In the prevention of chronic respiratory morbidity following RSV infection, however, studies are needed to determine whether immunoprophylaxis will have a useful role and to identify which drug treatment will be most cost-effective. PMID- 14521490 TI - Current treatment options to prevent perinatal transmission of herpes simplex virus. AB - Neonatal herpes is a potentially devastating consequence of perinatal transmission of the herpes simplex virus (HSV), with significant morbidity and mortality. Treatment options are available, but must begin early in disease with manifestations that are often protean. Thus, preventive measures need to be optimised. Antiviral suppression in late pregnancy of women with a history of recurrent genital herpes will decrease symptomatic recurrence at delivery and appears to reduce caesarian section rates. However, primary HSV Type 2 and primary HSV Type 1 episodes have the highest neonatal transmission rates and thus, effective prevention may require the identification and suppression of the discordant partner. Significant experience has been gained with the use of acyclovir in pregnancy and it is recommended for both episodic and suppressive therapy in pregnant women. Its use has been demonstrated to be cost-effective in suppressive therapy, although issues regarding compliance and the potential for neonatal neutropenia need to be addressed. The more conveniently dosed prodrugs valacyclovir and famciclovir are being evaluated for use in pregnancy. PMID- 14521491 TI - Current pharmacotherapy for the treatment of chronic hepatitis B. AB - The virological profile of infection with the hepatitis B virus (HBV) is changing in many parts of the world from the classical hepatitis B e antigen (HBeAg) positive serological pattern to a HBeAg-negative pattern, linked to the replacement of wild-type HBV by HBV variants with mutations in the core-promoter and in the precore region that prevent the secretion of HBeAg. The wild-type HBV disease is characterised by steady levels of alanine aminotransferase (ALT) and high HBV-DNA levels, responding relatively well to IFN treatment (3 - 5 MU/day or 10 MU every other day for 16 weeks), which induces anti-HBe seroconversion and normalises ALT levels in approximately 30% of the adults, with a minimal risk of relapse. Pegylated-IFN appears to have superior efficacy over conventional IFN alpha. Mutant-type disease (anti-HBe-positive/HBeAg-negative) is less responsive to IFN given for 6 - 12 months. This has led to the use of novel nucleoside analogues, of which the prototype is lamivudine. The response to lamivudine therapy shares with IFN a rapid decline in ALT accompanied by an improvement in histology; at variance with IFN, in HBeAg-positive chronic hepatitis B (CHB) there is delayed seroconversion to anti-HBe which accumulates over time, the switch to anti-HBs is more rare and in the long-term, the activity of the drug is abolished by the emergence of viral mutations (YMDD-motif mutants) that may rekindle the disease. The combination of IFN plus lamivudine may be more efficacious than IFN or lamivudine monotherapy. Lamivudine therapy needs to be prolonged in HBeAg-negative CHB. Short-term lamivudine-therapy is highly efficacious in preventing HBV reinfection in liver transplants. Recent data suggest that long-term IFN therapy (24 months) may achieve a response in 30% of HBeAg-negative patients. The advent of adefovir, an analogue of adenosine monophosphate, may provide a safer alternative to lamivudine in the control of HBV disease; the drug is well-tolerated and treatment raises drug-resistant mutants in < 2% of the patients over 2 years of therapy. Adefovir provides rescue therapy against YMDD mutants raised by lamivudine therapy. PMID- 14521492 TI - Paroxetine in the treatment of post-traumatic stress disorder: pooled analysis of placebo-controlled studies. AB - Post-traumatic stress disorder (PTSD) is increasingly understood to be a medical disorder characterised by particular psychobiological dysfunctions that respond to specific treatments. Paroxetine is a selective serotonin re-uptake inhibitor that has been found effective in the treatment of major depression as well as a range of anxiety disorders. This paper reviews data on the use of paroxetine for the treatment of adult PTSD. There have been three 12-week, placebo-controlled studies of paroxetine in PTSD. As these followed a partly similar design, a pooled analysis of the studies is possible and is reported here. Paroxetine is effective in the short-term treatment of PTSD, resulting in significantly better response and remission rates than placebo, improving sleep disturbance and reducing each of the symptom clusters of PTSD, as well as the disability associated with this condition. The medication is effective in both male and female PTSD patients and whether or not there are comorbid disorders such as depression. PMID- 14521494 TI - Gatifloxacin in community-acquired respiratory tract infection. AB - Gatifloxacin (Tequin, Kyorin Pharmaceutical Co. Ltd) is a new fluoroquinolone with a broad spectrum of activity for pathogens implicated in community-acquired respiratory tract infections, including Gram-positive, -negative and atypical bacteria. Excellent oral bioavailability, a half-life allowing once-daily administration and excellent penetration into respiratory tissues are desirable pharmacokinetic characteristics of gatifloxacin. Monte Carlo simulation of gatifloxacin in Streptococcus pneumoniae infection demonstrates that adequate levels of pharmacokinetic/pharmacodynamic parameters are obtained with gatifloxacin in almost all instances. In randomised, controlled trials and a large open-label, community-based study, gatifloxacin has shown excellent clinical and bacteriological efficacy in acute bacterial sinusitis, acute exacerbations of chronic bronchitis and community-acquired pneumonia. Current guidelines recommend the initial empiric use of gatifloxacin (along with the other new fluoroquinolones) for community-acquired respiratory tract infection in patients who have increased likelihood of infection with resistant pathogens. Another group of patients where this agent is recommended for initial antimicrobial therapy are those who, because of underlying disease and/or comorbid conditions, need an antibiotic with high antimicrobial efficacy to achieve optimal outcomes. PMID- 14521493 TI - Amoxicillin/clavulanate potassium extended release tablets: a new antimicrobial for the treatment of acute bacterial sinusitis and community-acquired pneumonia. AB - Community-acquired bacterial respiratory tract infections are among the most common health disorders requiring medical care and are associated with substantial morbidity, mortality, and direct and indirect costs. Recent increases in the prevalence of antimicrobial resistance have resulted in reduced susceptibility of the most common respiratory tract bacterial pathogens to a number of antimicrobials. Amoxicillin/clavulanate potassium extended release (ER) tablets (Augmentin XR, GlaxoSmithKline) is a new formulation of amoxicillin/clavulanate that retains activity against betalactamase-producing organisms whilst increasing the activity against Streptococcus pneumoniae through elevated and sustained plasma amoxicillin concentrations. The bilayer tablet provides immediate release of clavulanate and both immediate and sustained release of amoxicillin to maintain therapeutic concentrations of amoxicillin over longer periods of the dosing interval. In clinical trials of acute bacterial sinusitis (ABS) and community-acquired pneumonia (CAP), amoxicillin/clavulanate ER was shown to have excellent bacteriological and clinical success rates, even in patients infected with antimicrobial-resistant pathogens, and was found to be generally well tolerated. Amoxicillin/clavulanate ER is approved in the US for the treatment of patients with ABS or CAP caused by beta-lactamase-producing pathogens (ie, Haemophilus influenzae, Moraxella catarrhalis, Haemophilus parainfluenzae, Klebsiella pneumoniae, or methicillin-susceptible Staphylococcus aureus) and S. pneumoniae with reduced susceptibility to penicillin (penicillin minimum inhibitory concentration = 2.0 microg/ml). PMID- 14521499 TI - RE: Special notice! Binocul Vis Strabismus Q 2003;18:114. PMID- 14521495 TI - Is memantine a breakthrough in the treatment of moderate-to-severe Alzheimer's disease? AB - Over stimulation of the NMDA receptor by glutamate has long been considered to have a role in Alzheimer's disease (AD). Memantine, a non-competitive antagonist of NMDA receptors, has recently been tested in moderate-to-severe AD. At 28 weeks, there was a small significant mean difference of 0.3 in favour of memantine, showing benefit in the Clinician's Interview-Based Impression of Change Plus Caregiver Input (CIBIC-Plus) global score. The other primary efficacy variable was the Alzheimer's disease Cooperative Study Activities of Daily Living Inventory, modified for more severe dementia (ADCS-ADL-sev) and there was significantly less deterioration in the ADCSADL-sev score in the memantine group than in the placebo group (2.1 versus 3.4, respectively). As there are no drugs commonly used for their effectiveness in moderate-to-severe AD, any drug that causes a benefit, however small, as memantine has been shown to do in this trial, represents a breakthrough. PMID- 14521500 TI - RE: Reverse subspecialization? Is it time to separate ophthalmic pediatrics and strabology? Binocul Vis Strabismus Q 2003;18:11. PMID- 14521502 TI - Socialized medicine hooray? Second surgery for exotropia; 8 muscle surgery for nystagmus and strabismus; heroin ET; no surgery; monovision followup. PMID- 14521503 TI - Outcome of 5 mm resection of one medial rectus extraocular muscle for recurrent exotropia. AB - PURPOSE: To evaluate the success rate of 5 mm resection of one medial rectus (MR) as the second surgery for intermittent exotropia of childhood after bilateral lateral rectus recessions and after unilateral lateral rectus recessions, and to propose a theory of how this surgery works. CASES: The author performed a retrospective chart review of 10 years experience of routinely performing a 5 mm resection of one medial rectus as the second surgery for exotropia recurrent after unilateral or bilateral lateral rectus (LR) recession. A total of 45 cases of children receiving a second surgery for recurrent exotropia, a 5 mm resection of one medial rectus in all cases, were identified. Ten of the second surgeries were performed with the first surgery having been a large unilateral LR recession. Thirty-five had received bilateral LR recessions as the first procedure. Only those cases needing a third surgery prior to 2 years or with at least two years followup with continued good binocular alignment were included. OUTCOME: Residual deviations of 8 XT to 10 ET, inclusive, (or less) were considered surgical "successes". RESULTS: Of the 10 whose first surgery had been a unilateral LR recession (8-9 mm for 9 of the 10), the two year failure rate was 4/10. Of the 35 whose first surgery had been a bilateral LR recession (4.5 to 8.2 mm), the two year failure rate was 4/35. The time to failure after resection of one MR averaged 27 months for the 6/10 failures of the unilateral LR recession group; the time to failure after resection of one MR for the 14/35 failures of the bilateral LR recession group averaged 42 months. CONCLUSIONS: The more rapid failure of resection of one MR for those whose first surgery was a unilateral instead of a bilateral LR recession suggests that recession of the other LR may yield a higher success rate for the unilateral LR recession group in the future. Resection may work for a long time due to removal of embryonic myosin responsible for restoration of normal linear sarcomeric density. PMID- 14521504 TI - Acute esotropia in heroin withdrawal: a case series. AB - BACKGROUND: Esotropia during opiate withdrawal is a new clinical syndrome that has only recently been reported in the literature. METHODS: Clinical case series. RESULTS: Five patients with acute esotropia during opiate withdrawal are presented. In four there was evidence of underlying hyperopia and/or other strabismogenic features. CONCLUSIONS: The precise cause of esotropia with heroin withdrawal is uncertain. Most (and possibly all) of these patients have one or more objective strabismogenic features, and these are probably a large factor in the causation. PMID- 14521505 TI - When the patient says no. Management of exotropia with hemianopic visual field defects. AB - BACKGROUND: A homonymous hemianopia can be compensated for by an exotropia in the direction of the visual field defect. CASE REPORTS (AND LITERATURE REVIEW): Two young males with visual field defects and exotropia are reported. Both refused surgery when they were advised of the risk of reduction of their binocular visual field with the alignment of their eyes. CONCLUSION: Mapping of the binocular visual field is mandatory in patients with exotropia and neurological involvement to search for this rare combination which could produce a disappointing surgical outcome. PMID- 14521506 TI - Protective effects of the angiotensin II type 1 (AT1) receptor blockade in low renin deoxycorticosterone acetate (DOCA)-treated spontaneously hypertensive rats. AB - The present study evaluates the participation of oxidative stress, tissue angiotensin II (Ang II) and endothelin (ET) in the effects of losartan on blood pressure (BP), ventricular hypertrophy and renal injury in spontaneously hypertensive rats (SHRs), and explores how these effects are modified when spontaneous hypertension is transformed in a low-renin model by the administration of deoxycorticosterone acetate (DOCA). The following groups were used: SHR-control, SHR+DOCA, SHR+losartan and SHR+DOCA+losartan. Tail systolic BP was measured once a week. After 9 weeks of treatment, direct BP and metabolic, morphological, biochemical and renal variables were measured. DOCA administration to SHRs produced an increase in BP, ventricular hypertrophy, renal weight, proteinuria, renal histopathological lesions, urinary excretion of isoprostane F2alpha and ET levels in the renal cortex. Losartan reduced BP, plasma malondialdehyde levels, urinary excretion of isoprostane F2alpha, renal Ang II and renal and urinary levels of ET in the SHR and DOCA-treated SHR groups. Losartan increased plasma nitrite/nitrate in SHRs, but not in low-renin DOCA treated SHRs. Losartan reduced ventricular hypertrophy and ventricular Ang II in SHRs, but not in DOCA-treated SHRs. Losartan significantly decreased proteinuria and renal injury in DOCA-treated SHRs. We conclude that (i) the DOCA-induced aggravation of hypertension, ventricular hypertrophy and renal injury in SHRs is accompanied by augmented oxidative stress and increased levels of ET in the renal cortex, which could contribute to their development; and (ii) losartan reduced oxidative stress and renal Ang II and ET in SHRs and DOCA-treated SHRs, which might contribute to its antihypertensive and renoprotective effects, regardless of renin status. PMID- 14521507 TI - Influence of controlled breathing patterns on cerebrovascular autoregulation and cardiac baroreceptor sensitivity. AB - Transfer function analysis has become one of the main techniques to study the dynamic relationship between cerebral blood flow and arterial blood pressure, but the influence of different respiratory rates on cerebral blood flow has not been fully investigated. In 14 healthy volunteers, middle cerebral artery blood flow velocity, recorded using transcranial Doppler ultrasound, non-invasive beat-to beat Finapres blood pressure, ECG and end-tidal CO(2) ( P ECTO(2)) levels were recorded with subjects resting supine and breathing spontaneously or at controlled rates of 6, 10 and 15 breaths/min. Transfer function analysis and impulse and step responses were computed at each respiratory rate. P ECTO(2) levels tended to fall slightly during paced respiration, especially at 15 breaths/min. Controlled breathing rates did not alter transfer function analysis in the frequency range below 0.08 Hz but, above this frequency, the coherence function contained significant peaks corresponding to the respiratory frequencies. The impulse response was similar at all breathing rates, but the step response was characteristic of more efficient autoregulation with reduced P ECTO(2) levels associated with increasing respiratory rate. The effects of breathing rate and rhythmicity and P ECTO(2) must be considered in studies of cerebral autoregulation. PMID- 14521508 TI - Rho GTPases have diverse effects on the organization of the actin filament system. AB - The Rho GTPases are related to the Ras proto-oncogenes and consist of 22 family members. These proteins have important roles in regulating the organization of the actin filament system, and thereby the morphogenesis of vertebrate cells as well as their ability to migrate. In an effort to compare the effects of all members of the Rho GTPase family, active Rho GTPases were transfected into porcine aortic endothelial cells and the effects on the actin filament system were monitored. Cdc42, TCL (TC10-like), Rac1-Rac3 and RhoG induced the formation of lamellipodia, whereas Cdc42, Rac1 and Rac2 also induced the formation of thick bundles of actin filaments. In contrast, transfection with TC10 or Chp resulted in the formation of focal adhesion-like structures, whereas Wrch-1 induced long and thin filopodia. Transfection with RhoA, RhoB or RhoC induced the assembly of stress fibres, whereas Rnd1-Rnd3 resulted in the loss of stress fibres, but this effect was associated with the formation of actin- and ezrin-containing dorsal microvilli. Cells expressing RhoD and Rif had extremely long and flexible filopodia. None of the RhoBTB or Miro GTPases had any major influence on the organization of the actin filament system; instead, RhoBTB1 and RhoBTB2 were present in vesicular structures, and Miro-1 and Miro-2 were present in mitochondria. Collectively, the data obtained in this study to some extent confirm earlier observations, but also allow the identification of previously undetected roles of the different members of the Rho GTPases. PMID- 14521509 TI - Structural and kinetic properties of a novel purple acid phosphatase from phosphate-starved tomato (Lycopersicon esculentum) cell cultures. AB - An intracellular acid phosphatase (IAP) from P(i)-starved (-P(i)) tomato ( Lycopersicon esculentum ) suspension cells has been purified to homogeneity. IAP is a purple acid phosphatase (PAP), as the purified protein was violet in colour (lambda(max)=546 nm) and was insensitive to L-tartrate. PAGE, periodic acid Schiff staining and peptide mapping demonstrated that the enzyme exists as a 142 kDa heterodimer composed of an equivalent ratio of glycosylated and structurally dissimilar 63 (alpha-subunit) and 57 kDa (beta-subunit) polypeptides. However, the nine N-terminal amino acids of the alpha- and beta-subunits were identical, exhibiting similarity to the deduced N-terminal portions of several putative plant PAPs. Quantification of immunoblots probed with rabbit anti-(tomato acid phosphatase) immune serum revealed that the 4-fold increase in IAP activity due to P(i)-deprivation was correlated with similar increases in the amount of antigenic IAP alpha- and beta-subunits. IAP displayed optimal activity at pH 5.1, was activated 150% by 10 mM Mg(2+), but was potently inhibited by Zn(2+), Cu(2+), Fe(3+), molybdate, vanadate, fluoride and P(i). Although IAP demonstrated broad substrate selectivity, its specificity constant ( V (max)/ K (m)) with phosphoenolpyruvate was >250% greater than that obtained with any other substrate. IAP exhibited significant peroxidase activity, which was optimal at pH 9.0 and insensitive to Mg(2+) or molybdate. This IAP is proposed to scavenge P(i) from intracellular phosphate esters in -P(i) tomato. A possible secondary IAP role in the metabolism of reactive oxygen species is discussed. IAP properties are compared with those of two extracellular PAP isoenzymes that are secreted into the medium of -P(i) tomato cells [Bozzo, Raghothama and Plaxton (2002) Eur. J. Biochem. 269, 6278-6286]. PMID- 14521510 TI - The production of hypericins and hyperforin by in vitro cultures of St. John's wort (Hypericum perforatum). AB - St. John's wort ( Hypericum perforatum L.) is a herbaceous perennial distributed throughout the World that has been widely used in traditional medicine. H. perforatum produces several types of biologically active compound, including the hypericins--a family of light-activated anthraquinones, localized within specialized glands found predominantly on flowers and leaves--and the hyperforins -a family of prenylated acylphloroglucinols localized in the reproductive structures of the plant. Hypericins are known to be toxic to mammals and display antiviral and anticancer activity, but the role of these compounds within the plant is unknown. Hyperforins display potent antimicrobial activity and are thought to be the primary bioactive ingredient for anti-depressive effects of the herb. The introduction of H. perforatum from Europe into the U.S.A. occurred in the 17th Century. Since the plant is considered a noxious weed, few efforts have been carried out to analyse populations in the context of secondary-metabolite concentrations. But in terms of secondary-metabolite studies, H. perforatum is an ideal model system to study the biosyntheses of aromatic polyketides and regulation of those pathways by environmental and genetic influences. This is due, in part, to the ease of conducting these studies in plant tissue culture. This review describes the progress of secondary-metabolite studies currently underway using H. perforatum. Specifically, this Review focuses on the production and regulation of the hypericins and the hyperforin in wild populations, field cultivation, greenhouse studies and plant tissue culture. Additionally, factors optimizing compound production--particularly in in vitro cultures--are presented and reviewed. PMID- 14521512 TI - Mammary epithelial stem cells: transplantation and self-renewal analysis. AB - An entire mammary epithelial outgrowth, capable of full secretory differentiation, may be comprised of the progeny of a single cellular antecedent. This conclusion is based upon the maintenance of retroviral insertion sites within the somatic DNA of successive transplant generations derived from a single mammary fragment. In addition, dissociation of these clonal dominant glands and implantation of dispersed cells at limiting dilution demonstrated that both duct limited and lobule-limited outgrowths were developed, as well as complete, fully differentiated glands. Thus, transplantation has revealed three distinct mammary epithelial progenitors in the mouse. Similar studies have extended this observation to rat mammary tissue. Recently, using cre-lox conditional activation of reporter genes, a new epithelial progenitor, specific for mammary secretory epithelium in postlactation females has been uncovered. In situ, these cells were shown to regenerate secretory lobules upon successive pregnancies. In transplant studies, they demonstrated the capacity for self-renewal and contributed to the new generation of all of the known epithelial cell types among mammary epithelium. In limiting dilution, the parity-induced progenitors were capable of engendering lobule-limited and duct-limited outgrowths in their entirety, but not completely developed glands. Serial transplant studies indicate that these progenitors have a significant but limited capacity for self-renewal. PMID- 14521513 TI - Isolation and characterization of functional mammary gland stem cells. AB - Significant advances in the stem-cell biology of several tissues, including the mammary gland, have occurred over the past several years. Recent progress on stem cell fate determination, molecular markers, signalling pathways and niche interactions in haematopoietic, neuronal and muscle tissue may provide parallel insight into the biology of mammary epithelial stem cells. Taking advantage of approaches similar to those employed to isolate and characterize haematopoietic and epidermal stem cells, we have identified a mammary epithelial cell population with several stem/progenitor cell qualities. In this article, we review some recent data on mammary epithelial stem/progenitor cells in genetically engineered mouse models. We also discuss several potential molecular markers, including stem cell antigen-1 (Sca-1), which may be useful for both the isolation of functional mammary epithelial stem/progenitor cells and the analysis of tumour aetiology and phenotype in genetically engineered mouse models. In different transgenic mammary tumour models, Sca-1 expression levels, as well as several other putative markers of progenitors including keratin-6, possess dramatically altered expression profiles. These data suggest that the heterogeneity of mouse models of breast cancer may partially reflect the selection or expansion of different progenitors. PMID- 14521514 TI - Epithelial progenitor cell lines as models of normal breast morphogenesis and neoplasia. AB - The majority of human breast carcinomas exhibit luminal characteristics and as such, are most probably derived from progenitor cells within the luminal epithelial compartment. This has been subdivided recently into at least three luminal subtypes based on gene expression patterns. The value of knowing the cellular origin of individual tumours is clear and should aid in designing effective therapies. To do this, however, we need strategies aimed at defining the nature of stem and progenitor cell populations in the normal breast. In this review, we will discuss our technical approach for delineating the origin of the epithelial cell types. A major step forward was the purification of each cell type by the application of immunomagnetic cell sorting based on expression of lineage-specific surface antigens. We then developed chemically defined media that could support either the luminal epithelial or the myoepithelial cell phenotype in primary cultures. Having succeeded in continuous propagation presumably without loss of markers, we could show that a subset of the luminal epithelial cells could convert to myoepithelial cells, signifying the possible existence of a progenitor cell population. By combining the information on marker expression and in situ localization with immunomagnetic sorting and subsequent immortalization, we have identified and isolated a cytokeratin 19-positive suprabasal putative precursor cell in the luminal epithelial compartment and established representative cell lines. This suprabasal-derived epithelial cell line is able to generate both itself and differentiated luminal epithelial and myoepithelial cells, and in addition, is able to form elaborate terminal duct lobular unit (TDLU)-like structures within a reconstituted basement membrane. As more than 90% of breast cancers arise in TDLUs and more than 90% are also cytokeratin 19-positive, we suggest that this cell population contains a breast cancer progenitor. PMID- 14521515 TI - Regulation of human breast epithelial stem cells. AB - Breast epithelial stem cells are thought to be the primary targets in the aetiology of breast cancer. As breast cancers are predominantly oestrogen and progesterone receptor-positive (ERalpha/PR+), we investigated the biology of ERalpha/PR+ cells and their relationship to stem cells in normal human breast epithelium. Several complementary approaches were used to characterize the stem cell population and relate it to ERalpha/PR+ cells, including dual label colocalization on tissue sections, isolation of a Hoechst dye-effluxing 'side population' using flow cytometry, and examination of DNA label retention. The intermediate or suprabasal population suggested by others to be breast stem cells comprises ERalpha/PR+ cells that coexpress the putative stem-cell markers including cytokeratin 19. Human breast epithelial cells with Hoechst dye effluxing 'side population' properties characteristic of mammary stem cells in mice were demonstrated by lack of expression of myoepithelial and luminal cell specific antigens such as CALLA and MUC1 to be undifferentiated cells. Using DNA radiolabelling of human tissue implanted into athymic nude mice, a population of label-retaining putative stem cells (LRC) were shown to be enriched for cells expressing the putative stem-cell markers p21CIP1/WAF1 and Musashi-1, which, interestingly, were expressed in separate subpopulations of ERalpha/PR+ cells. Finally, expression patterns of Musashi-1 and Notch-1 in relation to ERalpha/PR+ and adjacent proliferating cells suggest that the evolutionarily conserved Delta/Notch signalling pathway regulates asymmetric division of the putative stem cell population. The data suggest a model in which ERalpha/PR+ cells scattered through the epithelium are stem cells that self-renew through asymmetric cell division and generate patches of transit amplifying and differentiated cells. ERalpha/PR+ breast cancers exhibit loss of the two key regulators of asymmetric cell division, Musashi-1 and Notch-1 and thus may arise from symmetric division of the ERalpha/PR+ stem cell. PMID- 14521516 TI - Stem cells in normal breast development and breast cancer. AB - The main focus of this review is the role of mammary stem cells in normal breast development and carcinogenesis. We have developed a new in vitro culture system that permits, for the first time, the propagation of mammary stem and progenitor cells in an undifferentiated state, which should facilitate the elucidation of pathways that regulate normal mammary stem-cell self-renewal and differentiation. Furthermore, we propose a model in which transformation of stem cells, or early progenitor cells, results in carcinogenesis. A key event in this process is the deregulation of normal self-renewal in these cells. Transformed mammary stem or progenitor cells undergo aberrant differentiation processes that result in generation of the phenotypic heterogeneity found in human and rodent breast cancers. This phenotypic diversity is driven by a small subset of mammary tumour stem cells. We will discuss the important implications of this mammary tumour stem-cell model. PMID- 14521517 TI - Evidence of progenitor cells of glandular and myoepithelial cell lineages in the human adult female breast epithelium: a new progenitor (adult stem) cell concept. AB - Although experimental data clearly confirm the existence of self-renewing mammary stem cells, the characteristics of such progenitor cells have never been satisfactorily defined. Using a double immunofluorescence technique for simultaneous detection of the basal cytokeratin 5, the glandular cytokeratins 8/18 and the myoepithelial differentiation marker smooth muscle actin (SMA), we were able to demonstrate the presence of CK5+ cells in human adult breast epithelium. These cells have the potential to differentiate to either glandular (CK8/18+) or myoepithelial cells (SMA+) through intermediary cells (CK5+ and CK8/18+ or SMA+). We therefore proceeded on the assumption that the CK5+ cells are phenotypically and behaviourally progenitor (committed adult stem) cells of human breast epithelium. Furthermore, we furnish evidence that most of these progenitor cells are located in the luminal epithelium of the ductal lobular tree. Based on data obtained in extensive analyses of proliferative breast disease lesions, we have come to regard usual ductal hyperplasia as a progenitor cell-derived lesion, whereas most breast cancers seem to evolve from differentiated glandular cells. Double immunofluorescence experiments provide a new tool to characterize phenotypically progenitor (adult stem) cells and their progenies. This model has been shown to be of great value for a better understanding not only of normal tissue regeneration but also of proliferative breast disease. Furthermore, this model provides a new tool for unravelling further the regulatory mechanisms that govern normal and pathological cell growth. PMID- 14521518 TI - Heart valve and arterial tissue engineering. AB - In the industrialized world, cardiovascular disease alone is responsible for almost half of all deaths. Many of the conditions can be treated successfully with surgery, often using transplantation techniques; however, autologous vessels or human-donated organs are in short supply. Tissue engineering aims to create specific, matching grafts by growing cells on appropriate matrices, but there are many steps between the research laboratory and the operating theatre. Neo-tissues must be effective, durable, non-thrombogenic and non-immunogenic. Scaffolds should be bio-compatible, porous (to allow cell/cell communication) and amenable to surgery. In the early days of cardiovascular tissue engineering, autologous or allogenic cells were grown on inert matrices, but patency and thrombogenicity of grafts were disappointing. The current ethos is toward appropriate cell types grown in (most often) a polymeric matrix that degrades at a rate compatible with the cells' production of their own extracellular matrical proteins, thus gradually replacing the graft with a living counterpart. The geometry is crucial. Computer models have been made of valves, and these are used as three-dimensional patterns for mass-production of implant scaffolds. Vessel walls have integral connective tissue architecture, and application of physiological level mechanical forces conditions bio-engineered components to align in precise orientation. This article reviews the concepts involved and successes achieved to date. PMID- 14521519 TI - Comparison of cell proliferation in the centre and advancing fronts of oral squamous cell carcinomas using Ki-67 index. AB - The cell proliferation status of 60 oral squamous cell carcinomas from Sri Lankan subjects was examined by immunohistochemistry using the Ki-67 index. A comparison was made between the indices derived from the centre of the tumours and those derived from the invasive fronts of the same tumours. There was a positive correlation between the two indices suggesting a clonal expansion of malignant cells, but the mean index derived for the invasive fronts (29.75 11.64) was significantly higher than the mean index for the body of these tumours (25.65 11.64). Thus, at a given time, more peripheral cells at the invasive front are proliferating and this compartment is likely to be more informative in prognostic and other behavioural studies involving the cell cycle. In squamous carcinomas, increased and uncontrolled cell proliferation at the invasive front may be one feature contributory to the invasion. PMID- 14521520 TI - The influence of STAT5 antisense oligonucleotides on the proliferation and apoptosis of selected human leukaemic cell lines. AB - The signal transducers and activators of transcription--STAT5A and STAT5B--take part in the regulation of many essential physiopathological processes. They influence the cell cycle, apoptosis and the proliferation of different types of cell lines. The STAT5 proteins are induced in response to multiple haematopoietic cytokines. Because they are constitutively active in certain haemato-oncologic diseases, it is also suggested that they play an important role in leukaemogenesis. However, function of these proteins in haematopoietic cell transformation and proliferation is not clear. The aim of this study was to evaluate the impact of perturbation of STAT5 expression [using oligodeoxynucleotide (ODN) against STAT5 mRNA], on the clonogenicity and survival of selected human leukaemic cell lines, HEL, HL-60, K562, TF-1. We analysed the effect of ODN pre-treatment on the cell clonogenicity in methylcellulose cultures according to the time and the temperature of exposure. Moreover, we attempted to estimate apoptosis induced in examined cells, by flow cytometry using combined Annexin V-PI staining and the TUNEL method. We also applied the RT-PCR method to analyse Bax and Bcl-xL gene expression. We found that the perturbation of STAT5 expression with antisense oligonucleotides caused a decrease in the proliferative potential of human K562 and TF-1 cell lines. Also, we observed higher induction of apoptotic cell death in the K562 and TF-1 cells incubated with the antisense STAT5A ODNs. We did not notice any impact of ODNs on the HL-60 and HEL cells. Our studies using STAT5 antisense oligonucleotides showed that these proteins may be critical in the regulation of growth and apoptosis of some types of leukaemic blasts. PMID- 14521521 TI - N-acetylcysteine induces beneficial changes in the acinar cell cycle progression in the course of acute pancreatitis. AB - Oxygen free radicals (OFR) are produced in the course of acute pancreatitis (AP). In addition to injurious oxidative effects, they are also involved in the regulation of cell growth. The aim of the present study was to examine the relationship between the effectiveness of N-acetyl-l-cysteine (NAC) to prevent the generation of OFR and the changes in the cell-cycle pattern of acinar cells in the course of AP induced in rats by pancreatic duct obstruction (PDO). NAC (50 mg/kg) was administered 1 h before and 1 h after PDO. Flow-cytometric measurement of OFR generation in acinar cells was carried out using dihydrorhodamine as fluorescent dye. Plasma amylase activity, pancreatic glutathione (GSH) content and TNF-alpha plasma levels were also measured. The distribution of acinar cells throughout the different cell-cycle phases was analysed at different AP stages by flow cytometry using propidium iodide staining. NAC administration reduced the depletion of pancreatic GSH content and prevented OFR generation in acinar cells of rats with PDO-induced acute pancreatitis. As a result, AP became less severe as reflected by the significant improvement of hyper-amylasaemia and maintenance of plasma TNF-alpha levels at values not significantly different from controls were found. NAC administration inhibited progression of cell-cycle phases, maintaining acinar cells in quiescent state at early PDO times. The protection from oxidative damage by NAC treatment during early AP, allows the pancreatic cell to enter S-phase actively at later stages, thereby allowing acinar cells to proliferate and preventing the pancreatic atrophy provoked by PDO-induced AP. The results provide evidence that OFR play a critical role in the progression of acinar cell-cycle phases. Prevention of OFR generation of acinar cells in rats with PDO-induced AP through NAC treatment, not only protects pancreas from oxidative damage but also promotes beneficial changes in the cell cycle progression which reduce the risk of pancreatic atrophy. PMID- 14521522 TI - Rickets: an old form for a new century. PMID- 14521523 TI - Determinants of prognosis of acute transverse myelitis in children. AB - BACKGROUND: Acute transverse myelitis (ATM) is a severe disorder; recovery requires several months and often leaves neurologic residua. To determine what features of patients with acute transverse myelitis significantly influence prognosis, the authors reviewed reports of ATM in Japanese children published in the last 15 years (from 1987 to 2001). METHODS: The authors studied reports of 50 Japanese patients (17 boys, 26 girls, 7 children of unspecified sex; mean age +/- SD, 8.0 +/- 3.8 years). Acute-phase and demographic features including age, increased deep tendon reflexes, Babinski reflex, sex, preceding infection, decreased deep tendon reflexes, time course of peak neurologic impairment, treatment with prednisolone and/or high-dose methylprednisolone, and the day of illness when treatment was started were used as independent variables in a regression analysis. The dependent variable was long-term persistence of neurologic deficits. RESULTS: Younger patients and those without increased deep tendon reflexes or a Babinski reflex were more likely to have residual neurologic deficits such as paraplegia or tetraplegia, sensory loss and sphincter disturbance. No relationship was seen between prognosis and sex, preceding infections, decreased deep tendon reflexes, time course of peak neurologic impairment, treatment with prednisolone or high-dose methylprednisolone, or timing of treatment initiation. CONCLUSIONS: Age at onset and neurologic features were important for outcome prediction in ATM. Steroid therapy did not associate with better outcome. PMID- 14521525 TI - Early head cooling in newborn piglets is neuroprotective even in the absence of profound systemic hypothermia. AB - BACKGROUND: Selective head cooling in the newborn infant has been proposed as a neuroprotective treatment with a lower level of systemic adverse effect than that of systemic hypothermia. However, the efficacy is not confirmed as well as that of systemic hypothermia. In order to analyze the safety and efficacy of selective head cooling, 25 newborn piglets were randomly selected for either normothermic or hypothermic treatment. METHODS: Global hypoxic insult was induced by lowering the oxygen concentration to the maximal level to maintain the background electroencephalogram (EEG) voltage under 7 microV for 45 min. The core temperature of normothermic piglets was maintained between 38.5 degrees C and 39 degrees C, while prophylactic cooling was applied to the hypothermic piglets at the same time of the insult. Very mild systemic hypothermia by 1 degrees C was induced in addition to selective head cooling with 10 degrees C coolant temperature. Animals were killed for histopathological examination seven hours after the end of the insult. RESULTS: Two normothermic piglets died while all hypothermic piglets survived. Neuropathological findings were significantly severer in the normothermic group than in the hypothermic group. Intracranial pressure was significantly lower, and EEG recovery was significantly better in the hypothermic piglets. There was no significant difference in the lowest oxygen concentration, degrees of acidosis, blood lactate, and blood pressure between the groups, although heart rate was significantly lower in the hypothermic group. CONCLUSIONS: We have demonstrated that early head cooling was effective in preventing some of the earliest brain damage due to hypoxic insult even in the absence of profound systemic hypothermia. PMID- 14521524 TI - Serum KL-6 level in newborns with meconium aspiration syndrome. AB - BACKGROUND: It has been reported that serum KL-6 increases in babies with progressing chronic lung disease (CLD). However, there have been few reports assessing KL-6 in meconium aspiration syndrome (MAS). KL-6 was measured in neonates with respiratory diseases including MAS. METHODS: Thirty-eight neonates with respiratory disease were enrolled in the study. These patients were classified into three groups, 14 patients with respiratory distress syndrome (RDS), 14 with MAS, and 10 with transient tachypnea of the newborn (TTN). The control group consisted of 12 healthy neonates. KL-6 levels were measured 1 day (median) after the birth. In the RDS group, measurement was repeated just prior to 36 weeks' postmenstrual age. RESULTS: The levels of KL-6 were 116 +/- 40 U/mL in the RDS, 281 +/- 138 U/mL in the MAS, and 106 +/- 41 U/mL in the TTN groups. KL-6 levels were significantly higher in the MAS group than in the control group (134 +/- 71 U/mL; P < 0.01). In addition, the levels were significantly higher in those with severe MAS than those with mild MAS (P < 0.05). In patients with RDS, KL-6 increased in patients who developed CLD (P < 0.05), while KL-6 levels did not change in those who did not develop CLD. CONCLUSION: These data confirm the high level of KL-6 in CLD, and suggest that KL-6 is increased in MAS. PMID- 14521526 TI - Flexible endoscopy of aerodigestive tract in small infants. AB - BACKGROUND: Flexible endoscopy (FE) is a useful method for diagnosing airway problems. Congenital or acquired airway lesions in infants may lead to respiratory distress that requires comprehensive investigation and management. This study was designed to evaluate the use of FE in small infants. METHODS: Infants who had symptoms of respiratory distress and received FE when they were less than 1-year-old were studied and their medical history, diagnoses, interventions, and complications from FE were investigated. RESULTS: The study population consisted of 568 small infants (334 boys and 234 girls) who weighed 5.1 +/- 2.4 kg, and received FE when they were 4.5 +/- 3.6 months of age. Most patients (91.2%) received diagnostic FE and the remainder (8.8%) received therapeutic procedures. Stridor (38.0%) was the most common indication for FE and laryngomalacia (33.3%) was the most frequent finding. Synchronous FE diagnosis was found in 351 (61.8%) cases. No major complications associated with FE were found. CONCLUSION: Flexible endoscopy allows direct visualization of dynamic motion of the small aerodigestive tract. Laryngomalacia was the most common FE finding of respiratory distress in small infants. Synchronous FE lesions were frequently found in this young age group and it necessitated a thorough investigation of the entire aerodigestive tract. PMID- 14521527 TI - Height improvement by L-thyroxine treatment in subclinical hypothyroidism. AB - BACKGROUND: Subclinical hypothyroidism(SH) is most commonly an early stage of hypothyroidism. Although the condition may resolve or remain unchanged, within a few years in some patients overt hypothyroidism develops, with low free T4 levels as well as a raised thyroid stimulating hormone (TSH) level. Patients with SH may have subtle hypothyroid symptoms with mild abnormalities of serum lipoproteins and cardiac functions. L-thyroxine in a dosage that maintains serum TSH levels within the normal range is the preferred therapy in these patients. Although short stature is a well-known clinical sign of overt hypo-thyroidism,the effect of SH in growth is not well established. The aim of the present study is to show the effect of treatment on height in these patients. METHODS: In the present study, 2067 patients who were admitted to the Endocrinology Department of SSK Ankara Children's Hospital, Ankara, Turkey, with the complaint of short stature were evaluated and 39 were diagnosed with SH by thyrotropin releasing hormone stimulation test. The anthropometric data of the patients who were subdivided into two groups (prepubertal and pubertal) were analyzed before and after 6-12 months of L-thyroxine treatment. Growth velocity (GV) and GV standard deviation score(GVSDS) of the groups before and after the treatment were statistically analyzed. RESULTS: Both groups showed significant increases in GV and GVSDS by L thyroxine treatment. CONCLUSION: Patients with short stature have to be evaluated for SH in addition to other potential causes. L-thyroxine treatment in these patients provides significant improvement in height. PMID- 14521528 TI - Somatostatin infusion withdrawal in the diagnosis of childhood growth hormone deficiency. AB - OBJECTIVE: An evaluation of growth hormone (GH) testing for GH deficiency (GHD) in childhood is confounded by the lack of a world-wide consensus on the definition of GHD. Although a single GH test remains the most powerful biochemical tool in the evaluation of a child with growth failure, the test remains far from ideal. Withdrawal of somatostatin (SS) infusion is followed by a rebound rise of GH thought to be mediated by endogenous GH-releasing hormone (GHRH) function. This study was designed to compare the GH response to 90 min SS infusion in children with normal GH secretion versus children with GH deficiency. METHODS: Ten children with GHD and 10 healthy controls (NC) have been evaluated for GH response to somatostatin infusion withdrawal (SSIW) and compared with response of two provocative tests, glucagon plus propranolol test and L-Dopa test. All children received constant infusion of somatostatin for 90 min (3 microg/kg per h, Stilamin, Serono, Aubonne, Switzerland). In order to determine GH, blood samples were obtained 90 min before the SS infusion and 0, 15, 30, 45, 60, 75, and 90 min after the cessation of infusion. RESULTS: Growth hormone peak levels with SSIW were significantly lower in GH deficient children than in healthy children (2.5 +/- 1.2 ng/dL, vs 21.9 +/- 5.3 ng/dL, respectively, P < 0.01). No adverse effects were observed during or after somatostatin infusion. CONCLUSION: In the present study, SSIW elicited a significant GH rise in healthy children but not in children with GH deficiency. Although further controlled studies using more data are necessary to expand these findings, the results suggested that children with GH deficiency can be reliably discriminated from healthy children by SSIW. PMID- 14521529 TI - High-dose immunoglobulin therapy for Guillain-Barre syndrome in Japanese children. AB - BACKGROUND: Guillain-Barre syndrome (GBS) is an acute acquired demyelinating polyneuropathy, presumed to be immune-mediated. Intravenous immunoglobulin (IVIg) has been used to treat GBS and was found to be effective. However, a well controlled study of pediatric GBS has not been conducted in Japan. Therefore, to evaluate the efficacy of IVIg in the treatment of GBS, an open-labeled study was performed in pediatric patients. METHODS: Participants in the study were required to be younger than 15 years old, and diagnosed as having moderate or severe GBS. IVIg (400 mg/kg per day) was administered to patients for five consecutive days. Predefined outcome measures were defined on a seven-point scale of motor function (Hughes' functional grade [FG]). RESULTS: Eleven patients were treated with IVIg. The median time taken to improve by one grade on the FG scale was 10.0 days after initial treatment. Two weeks after initial treatment, 72.7% of patients treated with IVIg improved by one or more grades, and 36.4% improved by two or more grades, measured on the FG scale. After 4 weeks an improvement by one or more grades was observed in 81.8% of patients, and two or more grades in 63.6% of patients. These improvement rates were markedly greater than would occur with the natural course of GBS1. Adverse events (subjective symptoms or abnormal laboratory findings) were observed in four patients, although all were temporary and mild. CONCLUSIONS: The authors conclude that IVIg is a safe and effective treatment for childhood GBS, which shortens the time to recovery. PMID- 14521530 TI - Serum leptin concentrations in children with mild-to-moderate protein-energy malnutrition. AB - BACKGROUND: The aim of the present study was to clarify the relationship between nutritional anthropometric parameters and serum leptin concentrations in otherwise healthy children with mild-to-moderate protein-energy malnutrition (PEM) secondary to inadequate energy intake. METHODS: Eighty-one otherwise healthy children with poor appetite and inadequate energy intake were enrolled in the study. The anthropometric values were evaluated in all subjects. Fasting serum leptin concentrations were assessed. RESULTS: Of the 81 otherwise healthy children, 30 were found to have mild-to-moderate PEM. The control group consisted of 51 healthy children who had normal anthropometric values, even though they had inadequate energy intake. There was a significant difference in serum leptin concentrations between the two groups (P < 0.01). Furthermore, there was a positive correlation between mean serum leptin concentrations and percentage of standard weight for height in the patient and the control group (P < 0.05). Multiple linear regression analysis indicated that percentage of standard BMI was the best predictor for serum leptin concentrations (P < 0.05). CONCLUSION: In addition to BMI, percentage of standard weight for height is a valuable nutritional anthropometric parameter for serum leptin concentrations in PEM and inadequate energy intake. However in the present study, percentage of standard BMI was the best predictor for serum leptin concentrations. Serum leptin concentrations are low not only in severe PEM, but also in children with mild-to moderate PEM without chronic disease. PMID- 14521531 TI - Efficacy of zinc supplementation on the severity and duration of diarrhea in malnourished Turkish children. AB - BACKGROUND: Intervention trials have shown that zinc may be efficacious in treating acute diarrhea in children of developing countries. A double-blind placebo-controlled study was designed to evaluate the effects of zinc supplementation on the clinical course and duration of diarrhea in malnourished Turkish children. METHODS: The study group comprised 40 subjects with low zinc levels (Group 1a) and 52 subjects with normal zinc levels (Group 1b). The control group was also comprised of two subgroups: 36 subjects with low zinc levels (Group 2a) and 54 subject with normal zinc levels (Group 2b). Forty-three percent of children in the study group and 40% of controls had low serum zinc levels (<14 micromol/L), and 43% of subjects in both groups had very low serum zinc concentrations (<10 micromol/L). The study group were given 20 mg zinc per day for 10 days and the control group were given 750 mg glucose per day as a placebo for 10 days. RESULTS: The mean duration of diarrhea was shorter and the percentage of children with consistent diarrhea for more than 3-7 days was lower in the study subgroups than in the control subgroups. Prolonged diarrhea was present in 12% of children in the study group, and in 44% and 37% of children in the hypozincemic and normozincemic control subgroups, respectively. The was no significant difference among the four subgroups of children in the number of cases with post-enrollment diarrhea of a duration of>14 days. Stool frequency over the first 4 days after enrollment was lower in children in the study group. CONCLUSION: It was concluded that zinc supplementation in malnourished children with acute diarrhea may reduce the severity and duration of diarrhea, especially in children with low zinc levels. PMID- 14521532 TI - Discharge diagnoses in infants with apparent life-threatening event. AB - BACKGROUND: There are various identifiable diseases or conditions that can be associated with an apparent life-threatening event (ALTE) in infancy. The present study was carried out to investigate the etiology of ALTE based on the discharge diagnoses. METHODS: A protocol for the complete work-up used to examine the cause of ALTE was designed, and was carried out for 69 infants with ALTE. RESULTS: Gastroesophageal reflux disease (GERD) was demonstrated in 38 cases, and chronic gastric volvulus (CGV) was diagnosed in 21 (associated with GERD in 13 infants). Four cases of pertussis and two cases each of sepsis, laryngomalacia, respiratory immaturity, and premature ventricular contractions were found. Others cases presented with pneumonia, meningitis, intussuception, food allergy, epilepsy, and adenoid vegetations. In 12 of the 69 infants with ALTE, no abnormalities were found in any of the studies performed, including two patients with breath-holding spells. In 17 cases, two or more possibly contributing findings were diagnosed. CONCLUSION: It is important to recognize that gastroesophageal impairments such as GERD or CGV could be responsible for many incidences of ALTE in infancy. The authors recommend a trial be established to investigate the causes of ALTE, including gastroesophageal evaluations in all infants with ALTE. PMID- 14521533 TI - Evaluation of 80 children with prolonged fever. AB - BACKGROUND: Several studies have been published regarding the etiology and evaluation of a child with prolonged fever, however, the reasons for the prolonged fever have changed during the years. The present study aims to determine the causes of prolonged fever, to investigate the relationship of fever using some basic laboratory tests, and to establish guidelines for the approach in those children. METHODS: The charts of 80 out of 17490 hospitalized children who were seen between 1996 and 2001 with prolonged fever of longer than 2 weeks and unknown origin were reviewed in the university hospital of Izmir, Turkey. Their charts were evaluated in respect of age, sex, growth curves, educational level of their families, the duration and the magnitude of fever, causes of fever, and basic laboratory investigations such as white blood cell, blood smear, hemoglobin, erythrocyte sedimentation rate, and C-reactive protein. RESULTS: Forty-four (55.00%) were boys and 36 (45.00%) were girls. Forty-four children (55.00%) were aged between 1 month and 2 years, 21 (26.25%) were aged 3-6 years, seven (8.75%) were aged 7-10 years, and eight (10.00%) were older than 10 years. The mean age was 3.87 +/- 4.17 years (range 3 months-17 years). Forty-six children (57.50%) had a prolonged fever that had lasted from 15-30 days, 18 (22.50%) from 31-60 days, and 16 (20.00%) had fever lasting more than 60 days. Final diagnosis had been reached in 70 of the 80 children (87.50%). The most common causes were infection (47/80), followed by immune deficiency (6/80), collagen tissue disorder (5/80), neoplasia (2/80), and miscellaneous (10/80) such as central fever in three, diabetes insipidus in two, familial Mediterranean fever in two, Kawasaki disease, foreign body in the respiratory system, and Crohn disease in one patient each. Among the laboratory tests white blood cell count, hemoglobin level and blood smear distribution of infection group were statistically significant. CONCLUSIONS: The most common cause of fever of unknown origin remains infection. The proportion of collagen tissue disorders and neoplasia have been found to be decreased. Unusual reasons such as diabetes insipidus and foreign body in the respiratory system in the miscellaneous group have been detected. Age plays important role in the diagnosis of prolonged fever, while some basic laboratory tests might give clues in the evaluation and may suggest a diagnosis. PMID- 14521534 TI - Fatal neonatal mitochondrial cytopathy with disseminated fatty nodules in the liver. PMID- 14521535 TI - Ritscher-Schinzel or 3C syndrome, with heterochromatic iris. PMID- 14521536 TI - Right-sided Duane retraction syndrome associated with multiple malformations. PMID- 14521537 TI - Case of Kawasaki disease in NICU. PMID- 14521538 TI - Use of recombinant human DNase in a premature infant with recurrent atelectasis. PMID- 14521539 TI - Case of congenital lobar emphysema in an 18-month-old boy and review of earlier cases. PMID- 14521540 TI - A trip in the gastrointestinal system with an unusual exit: fantastic voyage II. PMID- 14521541 TI - Two cases of congenital cytomegalovirus infection associated with disseminated intravascular coagulation. PMID- 14521542 TI - Infantile Amyand's hernia. PMID- 14521544 TI - Ontogeny of congenital anomalies of the kidney and urinary tract, CAKUT. AB - Ectopic budding of the initial ureter from the Wolffian duct is the first ontogenic misstep that leads to many congenital anomalies of the kidney and urinary tract (CAKUT). The ectopia results in hypoplastic kidney, ectopia of ureterovesical orifice, urinary outflow obstruction and/or reflux. Recent studies on several mutant mouse models verified that ectopic ureteric budding indeed occurs prior to the formation of CAKUT. Often, the genes involved in navigating the site of ureteric budding also regulate later ontogenic processes of the kidney and other urinary tract systems. These additional functions of the genes underlie the wide spectrum of CAKUT, for example multicystic dysplastic kidney, megaureter and atretic ureter, because the genes regulate the morphogenesis of the many portions of the excretory system through their distinctive cellular functions. PMID- 14521545 TI - Pathogenesis of human renal dysplasia: an alternative scenario to the major theories. AB - Among the various congenital anomalies of the kidney and urinary tract (CAKUT), renal dysplasia may deserve the closest attention due to its frequency and clinical severity. However, the pathogenesis of this disease is still not well known. It is believed that renal dysplasia is caused by a nephron induction deficit due to ampullary inactivity, or an abnormal budding of the ureteric bud from the mesonephric duct. Renal dysplasia is frequently associated with urinary tract obstruction, suggesting an alternative mechanism whereby urinary retention causes abnormal kidney development. Multicystic dysplastic kidney (MCDK) and obstructive renal dysplasia (ORD) are two different phenotypes of dysplasia associated with urinary tract obstruction. From detailed morphological studies using fetal dysplastic kidneys, nephron induction with filtrating function occurs before cysts are formed. Moreover, three-dimensional reconstruction of dysplastic nephrons showed that cysts developed in the once-induced nephrons as the result of fluid retention in both MCDK and ORD. In utero urinary tract obstruction may cause urine retention in functioning nephrons and lead to multiple cyst formation in the nephrogenic zone. These findings are common to MCDK and ORD, although there may be a difference in the obstruction site. Expansion of cysts with tubular dilatation (cysts) disturbs the subsequent nephrogenesis by the deregulated expression of growth and transcription factors, and may contribute to the misdevelopment of fetal kidneys. PMID- 14521546 TI - Genetic disorders of human congenital anomalies of the kidney and urinary tract (CAKUT). AB - Congenital abnormalities of the kidney and urinary tract, CAKUT are common in humans, occurring at a frequency of approximately 1 in 500 fetal ultrasound examinations. CAKUT are major causes of chronic renal failure in infants and young children, but little is known about the molecular pathogenesis of these disorders. To date, several gene mutations have been identified as a cause of human CAKUT: these include PAX2, KAL, EYA1, AGTR2 and HNF-1beta. At present, there is only limited information regarding how mutations alter gene expression during development to cause some CAKUT. The most convincing information comes from the multiorgan malformation syndromes with specific gene mutations. However, these syndromes are relatively rare, and most CAKUT appear to occur in isolation. The goal of this review is to provide an overview of these genetic disorders for CAKUT. An understanding of the genetic aspects of human CAKUT will help to unravel the pathogenesis of these disorders and may facilitate the design of genetic screening tests for early diagnosis and appropriate genetic counseling. Moreover, a deeper insight into the relationship between abnormal genes and the pathogenesis of abnormalities of CAKUT will provide an etiological classification of CAKUT. In addition, the importance of developing a registry of patients with various forms of CAKUT is discussed. This information will allow us to combine molecular biology and classical epidemiologic methods, and to continue expanding our knowledge regarding CAKUT. PMID- 14521547 TI - Ultrasound screening for renal and urinary tract anomalies in healthy infants. AB - BACKGROUND: Nearly 30% of childhood cases of chronic renal failure in Japan are attributed to congenital anomalies of the kidney and urinary tract (CAKUT), and the number is increasing. Urine screening at school facilitates early diagnosis and treatment of glomerulonephritis, but early screening for anomalies is currently not in practice. The authors evaluated the value of early abdominal ultrasonography screening in 1-month-old infants. METHODS: The following characteristics of kidneys were assessed: presence versus absence, size, symmetry of size, position, separation of the central echo complex (CEC), abnormal echogenicity, and other abnormal findings. The bladder and ureter were checked for abnormalities in bladder shape and wall, as well as retrovesical ureteral dilation. Criteria for abnormalities included kidney length of or=60 mm; a difference in length of left and right kidneys of 10 mm or more; and CEC separation of Society for Fetal Urology (SFU) grade 2 or higher. RESULTS: Beginning in April 1994 and continuing until September 2001, screening of 5700 1 month-old infants yielded 198 positive cases (3.5%) of CAKUT. Most frequent was abnormal CEC separation (approximately 60% of all abnormalities), followed by abnormal renal size or size asymmetry (30%). Further investigation yielded a specific diagnosis in 32 cases (0.6%) of all subjects. Most prevalent was obstructive uropathy (15 cases); 8 children underwent surgery. Small kidneys and vesicoureteral reflux were next in frequency. CONCLUSIONS: Ultrasonograpy was effective for early detection of renal and urinary tract anomalies. Ultrasound screening in early infancy may permit early treatment, that can prevent renal dysfunction. PMID- 14521548 TI - Medical management of congenital anomalies of the kidney and urinary tract. AB - Renal damage in children has been found to be more congenital in origin than was previously thought. Congenital anomalies of the kidney and urinary tract (CAKUT) involve renal dysplasia, renal hypoplasia, urinary tract obstruction and vesicoureteral reflux. CAKUT are sometimes bilateral and different types often coexist. Depending on their types and severity, children with CAKUT often have varying degrees of a reduced number of nephrons at birth. CAKUTare now the leading cause of renal failure in children. Children with renal dysplasia or obstructive uropathy may have abnormal renal tubules, and tend to lose essential water and sodium in urine. This can lead to poor body growth unless they are supplemented with water and sodium. Children with severe ureteric reflux often develop urinary infection and renal scarring. Renal scarring can further increase the risk of renal failure in children who already have other CAKUTand fewer nephrons than normal. Hypertension and proteinuria may develop in children with renal dysplasia and further aggravate renal function unless they are treated. Recent advances in the understanding and management of CAKUT make it possible for children with CAKUT to grow normally, have fewer complications such as urinary infection, have longer renal survival, and survive even with end-stage renal diseases through renal replacement therapy. PMID- 14521549 TI - Changing concepts in urological management of the congenital anomalies of kidney and urinary tract, CAKUT. AB - Recent advancement in ultrasonographic evaluation has prompted early detection and diagnosis of congenital anomalies in the kidney and urinary tract (CAKUT) in the asymptomatic phase. Consequently, early surgical intervention has become possible in the asymptomatic phase for the purpose of controlling manifestations early, thereby avoiding renal functional deterioration. However, some lesions detected by ultrasonography have been shown to often resolve or disappear without intervention. Thus, it has become more important to identify and understand the natural history of CAKUT. For the precise evaluation of the results of surgical intervention, one must understand the maturational process of renal function during infancy. Without considering this process, we cannot differentiate the renal significance of the surgical management from the natural course of CAKUT. Recent advancement in the field of radioisotopic studies has also made a major contribution to the more precise assessment of renal function. Recent progress in the understanding of the pathophysiology and the natural history of CAKUT has helped rationalize its treatment and management. Improvement in the surgical techniques and tools, together with improvements in pediatric anesthesiology, have made an appreciably positive impact on the outcome. Herein, we present the emerging concepts in the urological management of CAKUT, specifically, multicystic dysplastic kidney, vesicoureteral reflux, congenital hydronephrosis, ectopic ureters and ureteroceles. PMID- 14521550 TI - Criteria for medical intervention in obese children: a new definition of 'obesity disease' in Japanese children. AB - The Committee of the Japan Society for the Study of Obesity reported the new criteria for 'obesity disease' for Japanese adults in 2000. We defined the criteria for the diagnosis of obesity in children with medical problems, corresponding to the 'obesity disease' criteria in adults. Obesity in childhood was defined as follows: percentage of overweight (POW) and body fat exceeded the criteria. 'Obesity disease in childhood' was defined as obesity associated with health or medical problems, and with indications for medical intervention. Medical problems with indications for immediate intervention were grouped as A problems, which consisted of (i). hypertension; (ii). sleep apnea or hypoventilation; (iii). Type 2 diabetes mellitus or impaired glucose tolerance; and (iv). increased waist circumference or accumulation of visceral adipose tissue. Metabolic derangements or equivalent associated with obesity were grouped as B problems: (i). liver dysfunction; (ii). hyperinsulinemia; (iii). hypercholesterolemia; (iv). hypertriglyceridemia; (v). low serum high-density lipoprotein cholesterol; (vi). acanthosis nigricans, and (vii). hyperuricemia. Obese children over 5 years of age with following conditions were diagnosed as 'obesity disease in childhood': (i). any 'A problem', (ii) POW >or= 50% and any 'B problem', or (3) POW < 50% and more than one 'B problem' or equivalent. We decided to take physicosocial problems related to obesity into consideration as the criteria. The resultant criteria are proposed by the Committee for Research of Appropriate Body Build in Children*. PMID- 14521552 TI - Two hundred ninety-six cases of onychomycosis in children and teenagers: a 10 year laboratory survey. AB - There is still little data available about the epidemiology of childhood onychomycosis. Looking at our laboratory figures over a 10-year period provided us with some useful information. Nail keratin samples were taken by dermatologists from 21,557 patients with nail conditions, mainly in the Brussels region. The specimens were examined by direct microscopy and/or histology, and cultured on Sabouraud medium agar. Only patients less than 17 years of age were considered as children. Clinical information was gathered about age, sex, and the location of the infected nail. Nine hundred sixty-three of the samples were from children, and 296 of those children had proven onychomycosis. More than three fourths of the cases were found in children more than 6 years old, and boys were more frequently affected than girls. Toenails were the predominant location of infection. Trichophyton rubrum was the main pathogen, followed by Candida spp. and Trichophyton interdigitale. One case was caused by Scopulariopsis spp. As in adults, onychomycosis is probably the main nail disease in children. After the age of 6 years, the presentation is very similar to that in adults: toenails are mostly involved, and T. rubrum, the main pathogen, is responsible for distal and lateral subungual onychomycosis. PMID- 14521551 TI - AQP1 and AQP3, psoriasin, and nitric oxide synthases 1-3 are inflammatory mediators in erythema toxicum neonatorum. AB - Erythema toxicum neonatorum is a common, inflammatory skin reaction in healthy newborn infants characterized by an accumulation of activated immune cells in the lesions. Its etiology and physiologic significance are still unclear. The purpose of this study was to extend the search for possible inflammatory mediators of the rash. We performed immunohistochemistry on punch biopsy cryosections from lesions of four, 1-day-old infants and from four matched controls without rash, using antibodies against the water channel proteins aquaporin-1 (AQP1) and aquaporin-3 (AQP3), psoriasin, and the nitric oxide synthase (NOS) enzymes, neuronal NOS (nNOS), inducible NOS (iNOS), and endothelial NOS (eNOS). All sections from the lesions showed a dense, nodular cellular infiltrate located near the hair follicle. The vessels in the dermis showed a high incidence of AQP1 and eNOS. Strong staining for AQP1, AQP3, and psoriasin, as well as nNOS, iNOS, and eNOS were seen in the entire epidermal layer. The infiltrate in the dermis contained numerous cells expressing AQP1, AQP3, nNOS, iNOS, and eNOS. Double immunofluorescence staining showed that AQP3 was located in CD1a-expressing Langerhans cells and other dendritic cells in the dermis, as well as in CD14 expressing macrophages, CD15-expressing neutrophils, and EG2-expressing eosinophils surrounding the hair follicle. Our findings show that AQP1 and AQP3, psoriasin, and NOSs are involved in the activation of the skin immune system at birth. PMID- 14521553 TI - Frequency and clinical role of Staphylococcus aureus overinfection in atopic dermatitis in children. AB - The goal of this study was to evaluate the frequency and role of Staphylococcus aureus infection in patients with atopic dermatitis (AD). In 81 children, ages 2 months to 9 years, affected with moderate to severe AD, 308 samples from the cutaneous lesions were obtained and analyzed. S. aureus was isolated in 52 children (64.2%). Five of these were also colonized by Streptococcus pyogenes and one by Candida albicans. In 61 patients, total IgE serum level and specific IgE were tested to evaluate their allergic status: in 43 children a diagnosis of extrinsic AD was made, while 18 were affected by intrinsic AD. A higher presence of the bacterium was observed in allergic (71%) versus nonallergic children (49%). Our data demonstrate the importance of S. aureus in the clinical manifestation of AD and, in particular, its role in worsening the eczematous lesions of the face, neck, and perineum in children less than 1 year of age. PMID- 14521554 TI - Contact sensitization to disperse dyes in children. AB - From January 1996 to December 2000, 1098 children, including 667 subjects with suspected allergic contact dermatitis and 431 patients with atopic dermatitis (AD), were patch tested with seven disperse dyes: disperse blue 124 (DB124), disperse blue 106 (DB106), disperse red 1 (DR1), disperse yellow 3 (DY3), disperse orange 3 (DO3), p-aminoazobenzene (PAAB), and p-dimethylaminoazobenzene (PDAAB). Of these, 51 patients (4.6%; 34 girls and 17 boys) proved sensitized to disperse dyes. AD or history of AD was present in 30 patients (59%). The most common sensitizer was DY3 (17 patients), followed by DO3 (15 patients), and DB124 (14 patients). Among dye-positive patients, about 12% were sensitized to disperse dyes alone and only 14% reacted to para-phenylenediamine. In disperse dye sensitive children not affected by AD, the feet, axillae, and groin appeared to be the most common localizations, whereas in those with AD, involvement of the face and the flexural areas of the limbs was more common. In conclusion, our study showed that in children with suspected contact sensitization, disperse dyes should be regarded as potential triggering allergens. PMID- 14521555 TI - Erythema dyschromicum perstans in prepubertal children. AB - Erythema dyschromicum perstans (EDP) is a rare disorder of pigmentation that is most common in Hispanic patients. In adults, EDP has a slow onset and is unlikely to resolve spontaneously. The etiology and clinical course in children is poorly defined. Physical examinations, chart reviews, and telephone interviews were performed for eight pediatric patients with EDP who were followed at Children's Memorial Hospital in Chicago between 1990 and 1998. All the patients available for long-term follow-up (five of the eight) experienced complete clearance without recurrence in an average of 2.5 years. In all of our patients, the onset was noted from July to December. The administration of aminopenicillins was coincident with the development of EDP in two of the patients. Review of the English-language literature reveals that 25 prepubertal children have previously been reported. Including our patients, 69% of prepubertal children with EDP experienced resolution. We concluded that the clinical course of childhood (prepubertal) EDP differs from that of adult EDP, and it is more likely to resolve within 2-3 years. PMID- 14521556 TI - A report of cutaneous tuberculosis in siblings. AB - Cutaneous tuberculosis is rarely seen in the United States, and we are unaware of any reports of siblings sharing the diagnosis. We report an instance of cutaneous tuberculosis affecting two sisters. The first sibling had skin biopsy specimen findings suggestive of an infectious source, but special stains and cultures were negative. Only with further evaluation using polymerase chain reaction (PCR) was the diagnosis established. The second sibling had positive staining for acid-fast bacilli, though her cultures were also negative. A high level of suspicion for tuberculosis should be maintained in otherwise healthy children with nonspecific, granulomatous disorders of the skin. PCR may be a useful tool for selected cases in which a high level of clinical suspicion exists, but special stains and cultures are negative. PMID- 14521557 TI - Tinea capitis due to Trichophyton soudanense in Cincinnati, Ohio, in internationally adopted children from Liberia. AB - Two sisters, ages 5 and 6 years, who were adopted from Liberia, presented in Cincinnati, Ohio, with tinea capitis. Fungal cultures grew Trichophyton soudanense, a dermatophyte rarely seen in North America. Both girls had multiple other infections. They responded well to therapy with oral griseofulvin and topical selenium sulfide. With the current increase in international travel and adoption, nonendemic dermatophyte infections such as this need to be recognized. PMID- 14521558 TI - Gangrenous cutaneous mucormycosis in a child with a solid organ transplant: a case report and review of the literature. AB - Infections by members of the Phycomycetes class of fungi typically arise in patients with substantial underlying immunosuppression. Several clinical presentations, including that of gangrenous cellulitis, have been described. In the pediatric population, critically ill premature newborns and older children with underlying hematopoietic malignancies and/or bone marrow transplantation are affected. Presentation in children with solid organ transplants is rare. Successful therapy for this condition includes aggressive debridement of infected tissue and systemic administration of amphotericin B. PMID- 14521559 TI - Sarcoidosis in a preschooler with only skin and joint involvement. AB - Sarcoidosis is a multisystemic granulomatous disease of unknown etiology. It is relatively rare in children less than 15 years of age and especially in those less than 5-6 years of age. Sarcoidosis characteristically involves the skin, eyes, and synovial tissues in patients less than 5-6 years of age. We report a 3 year-old boy with sarcoidosis who had cutaneous findings with joint symptoms. Dermatologic examination revealed lichenoid, erythematous, 2-3 mm papules, some of them grouped, all over his extremities and trunk. There were symmetric swellings on his ankles and wrists without erythema or pain which did not interfere with function. However, until now, no eye involvement had been detected in the patient. PMID- 14521560 TI - Andogsky syndrome variant: atopic dermatitis associated with bilateral cataracts and retinal degeneration with left retinal detachment. AB - A 15-year-old girl had a new variation of Andogsky syndrome (unilateral cataract and atopic dermatitis) in which atopic dermatitis was associated with primary bilateral cataracts and retinal degeneration with primary left retinal detachment before cataract surgery. She had no history of systemic steroid treatment. PMID- 14521561 TI - Harlequin ichthyosis in association with hypothyroidism and juvenile rheumatoid arthritis. AB - Harlequin ichthyosis is a rare and severe congenital erythrodermic ichthyosis characterized at birth by hyperkeratotic plates covering the entire body, ectropion, eclabium, poorly developed ears, and contractures of the hands and feet. Two Chinese children, a 2-year-old boy and an 11-year-old girl, presented with these classic features as well as alopecia and loss of eyebrows and eyelashes. The boy was small for his age and was found to have hypothyroidism at the age of 18 months; he is currently on thyroxine replacement therapy. At 6 years of age, the girl developed symmetrical polyarthritis associated with positive rheumatoid factor and radiologic evidence of erosive arthritis, suggestive of juvenile rheumatoid arthritis. She received prednisolone, nonsteroidal anti-inflammatory drugs (NSAIDs), and subsequently methotrexate for her arthritis, with clinical and radiologic improvement. Early therapy with oral retinoids in both children accelerated shedding of the hyperkeratotic plates as well as improved ectropion and eclabium. There was no major adverse reaction to oral retinoids. The development of juvenile rheumatoid arthritis in survivors with harlequin ichthyosis has not been previously described. The use of prednisolone and NSAIDs in the girl did not affect the skin condition, but the addition of methotrexate led to a decrease in erythema. The association with autoimmune disease is probably coincidental. The psychosocial impact of this severe lifelong disease on the two families was enormous. Early retinoid therapy may improve the disorder and help increase survival rates. A multidisciplinary approach, including psychosocial support of the affected families, is vital in the management of this lifelong disease. PMID- 14521562 TI - Leg ulcer in hereditary spherocytosis. AB - Indolent leg ulcers are a rare complication found in patients with hereditary spherocytosis. We report a 13-year-old girl with hereditary spherocytosis who developed a chronic painful ulcer on the medial malleolus. All other etiologies were ruled out. Nine months after splenectomy the ulcer healed completely and the symptoms disappeared. We discuss and review this unusual entity in children. PMID- 14521563 TI - Tuberculosis in a child presenting as asymptomatic oropharyngeal and laryngeal lesions. AB - We describe an 11-year-old boy who had several, asymptomatic, erythematous papules in the oropharynx and larynx with recent onset, two cervical lymphadenopathies, and a painless, erythematous plaque on the right wrist with a 2.5-year history of slow growth. Histologic examination of the mucocutaneous lesions revealed a submucous infiltrate of lymphocytes and Langhans giant cells in the papules and granulomatous dermatitis in the plaque. The cervical lymph node was biopsied and on the surgical scar, an erythematous, nodular lesion developed. A biopsy specimen of this lesion showed tuberculoid granulomas with prominent caseation necrosis, and culture was positive for Mycobacterium tuberculosis. The Mantoux test was strongly positive with a vesicular response. A diagnosis of mucocutaneous lupus vulgaris and scrofuloderma secondary to cervical tuberculous lymphadenitis was made. Two months after initiation of antituberculosis therapy there was a complete resolution of mucous lesions and healing with atrophic scars on the neck and wrist. This is a rare presentation in the literature and reminds clinicians that tuberculosis should be kept in mind in the differential diagnosis of oral cavity lesions. PMID- 14521564 TI - Intractable diarrhea of infancy with facial dysmorphism, trichorrhexis nodosa, and cirrhosis. AB - A 3-year-old girl had severe intractable diarrhea with trichorrhexis nodosa and cirrhosis. This patient was referred to the pediatric dermatology clinic for lifelong brittle hair. The brittle hair microscopically demonstrated trichorrhexis nodosa. The girl also had facial dysmorphism, with a prominent forehead and cheeks, broad flat nose, and hypertelorism. She had a history of severe intractable diarrhea since 2 weeks of age and failure to thrive requiring lifelong total parenteral nutrition (TPN). Hepatomegaly was noted and prompted liver biopsy which demonstrated cirrhosis. Mental retardation and developmental delay was also found upon examination. This child may be included in the syndrome of intractable infant diarrhea, an entity known in the gastroenterology literature but yet not reported in the dermatologic literature. Dermatologists should be aware of this syndrome in which trichorrhexis nodosa is commonly seen. PMID- 14521565 TI - Disseminated and disfiguring molluscum contagiosum in a child. AB - We report widely disseminated and disfiguring lesions of molluscum contagiosum occurring in a 4-year-old girl secondary to a primary immunodeficiency disorder. The child, born of consanguineous parents, had multiple, asymptomatic, raised, skin-colored and whitish lesions since 2 years of age. On physical examination she had growth retardation features with grade II protein energy malnutrition. Cutaneous examination revealed numerous, widespread, skin-colored to translucent, firm, umbilicated papules of varying sizes (pinpoint to larger than 10 mm). They were distributed bilaterally over all four limbs, including the dorsum of the hands and feet, trunk, face (including lips), eyelids, auriculae, and perineal and gluteal areas. In places, they were linearly disposed and had coalesced to form plaques. A biopsy specimen from a representative skin lesion (arm) demonstrated lobulated epidermal growth consisting of keratinocytes with large intracytoplasmic eosinophilic inclusion bodies and a central crater. These findings were consistent with the diagnosis of molluscum contagiosum. Topical therapies with phenol and trichloroacetic acid were ineffective The child succumbed to a fulminant systemic infection at home, 2 months after discharge from the hospital. The extent and distribution of molluscum contagiosum in our patient was remarkably vivid and has been hitherto reported only rarely. PMID- 14521566 TI - Imiquimod is highly effective for extensive, hyperproliferative condyloma in children. AB - We describe a dramatic response to imiquimod of long-lasting, highly proliferative extensive perianal condylomas involving the anal canal in a 19 month-old girl. Her mother was free of condyloma and allegedly had no human papillomavirus (HPV) infection during pregnancy. There was no evidence of sexual abuse. Application of 5% imiquimod cream to the child every other day for 3 weeks resulted in almost complete resolution of the warts, with total clearance within another 2 weeks. The inflammatory reaction was moderate. Since there is still discussion of whether imiquimod may be prescribed for small children, this case of very extensive condyloma provides evidence that the compound is safe and highly effective. PMID- 14521567 TI - Isotretinoin treatment of acne in a patient with Apert syndrome. AB - Apert syndrome is an autosomal dominant disease caused by mutations in fibroblast growth factor receptor (FGFR-2) and characterized by premature fusion of the cranial sutures and early epiphyseal closure. The development of widespread, severe, pustular acne around puberty is a characteristic feature in these patients. The FGFR-2 mutation of Apert syndrome has been described in localized nevoid acne lesions, which have been attributed to somatic mosaicism. Isotretinoin has been reported to be useful in controlling this acne. We report the successful treatment of acne with isotretinoin in a teenager with Apert syndrome and review the pathophysiology of acne in this syndrome. PMID- 14521568 TI - What syndrome is this? Costello syndrome. PMID- 14521569 TI - Multiple asymptomatic papules in a child. PMID- 14521571 TI - Erythema multiforme in a 9-day-old neonate. PMID- 14521570 TI - Giant shagreen patch associated with spina bifida occulta in tuberous sclerosis. PMID- 14521572 TI - Isolated nail dystrophy suggestive of dominant dystrophic epidermolysis bullosa. PMID- 14521573 TI - Acral pseudolymphomatous angiokeratoma of children (APACHE). PMID- 14521574 TI - Wet suit-related Pseudomonas aeruginosa dermatitis in a child. PMID- 14521575 TI - Huge keloid formation after BCG vaccination. PMID- 14521577 TI - Pfs230: from malaria transmission-blocking vaccine candidate toward function. PMID- 14521576 TI - Fibroblastic rheumatism: a variant of non-Langerhans cell histiocytoses? PMID- 14521578 TI - Influence of maternal infection on offspring immune response in murine larval toxocariasis. AB - The impact of Toxocara canis infection on the proportion of CD4+ and CD8+ T splenocytes, the serum concentrations of cytokine IFN-gamma and IL-5, and the production of Toxocara-specific antibodies were studied in two C57BL6/J mouse groups and their offspring. The mice were infected with 1000 T. canis eggs on the day of mating (early infection) and on day 14 of pregnancy (late infection). Early infection resulted in a significant increase of CD4+ T-cell subtype, however, a decline in CD8+ in comparison with late infection, as well as with non infected control. The IFN-gamma serum concentrations decreased in infected mothers after the birth when compared with non-infected mothers, while in the offspring this cytokine was barely or not detectable. In the mothers of both infected groups, the humoral immune response included both parasite-specific IgM and IgG2 antibodies. While IgG1 levels remained constant throughout the whole experiment in mothers with early infection, late-infected mothers became seropositive only 3 weeks after delivery. IgM was not detectable in any offspring. Pups from early-infected mothers had IgG1 antibodies. Conversely, IgG2 was detectable in pups of both experimental infection groups. A significant difference was observed in the amounts of pups/litter of the infected mothers in comparison with the non-infected ones. Only 56% of females after early infection and 79% of those after late infection had a successful pregnancy. However, all mice of the control group produced a litter. The first T. canis larvae were detected in the muscles of the offspring of both groups on day 5 after the birth. These data show the changes in regulatory and cytotoxic immunity mechanisms of the infected mothers and their offspring and the high level of pregnancy loss as a result of larval toxocariasis. PMID- 14521579 TI - Blockade of macrophage migration inhibitory factor (MIF) in Schistosoma japonicum infected mice results in an increased adult worm burden and reduced fecundity. AB - Macrophage migration inhibitory factor (MIF), a cytokine produced by many cell types, modulates cellular and humoral immune responses. In schistosomiasis, ova in the portal circulation induce a delayed type hypersensitivity (DTH) that results in formation of hepatic granulomas (HG) which secrete MIF activity. Therefore, we hypothesized that endogenous MIF modulates immune responses in schistosomiasis. To test this hypothesis, Schistosoma japonicum-infected mice were injected with rabbit IgG or neutralizing rabbit IgG antibody to MIF 4.5-6.5 week post infection when HG form and female worms are laying eggs. Compared with controls, 6.5-7-week post-infection, antibody-treated mice had 1.7-3 times as many adult worms and half as many ova per worm pair in their livers. In contrast, antibody introduced before infection or 6-8 week post infection did not affect worm burden or fecundity. Thus, for the first time there is evidence that 4.5-6 week post-infection endogenous MIF somehow mediates reduction of adult worm burden and promotes fecundity. Splenocytes and HG cells from antibody-treated mice showed reduced intracellular expression of TNFalpha and/or IL-10. We hypothesize that endogenous MIF enhances adult worm attrition by up-regulating innate and adaptive immune responses by increasing expression of MHC-II, co stimulatory, adhesion, receptor and cytokine molecules, and promotes fecundity by up-regulating TNFalpha expression. PMID- 14521580 TI - Local immune responses in sensitized sheep following challenge infection with Teladorsagia circumcincta. AB - Sheep were sensitized by weekly infections with Teladorsagia circumcincta over a 9-week period. After a 12-week rest, sheep were divided into four groups and killed without challenge or 3, 5 and 10 days post challenge (DPC) with 50000 L3. Recovery of challenge larvae from abomasal scrapings was highest at 3 DPC while no parasites were recovered by 10 DPC. Abomasal lymph nodes (ALN) of challenged sheep were significantly larger at 5 DPC, coinciding with an increase in the proportion of CD4 T cells and a decrease in CD21+ cells, probably reflecting the loss of CD21 from terminally differentiated antibody secreting cells. A significant increase was observed in gammadelta-TCR+ cells at 3 DPC in the ALN, while their number slightly decreased in the abomasal tissues throughout the challenge period. The number of tissue eosinophils was dramatically increased after challenge compared with the unchallenged controls, with a peak at 3 DPC, coinciding with the peak in larval recovery. CD4+ cells significantly increased in the abomasal tissues at 5 DPC, while no changes in globule leucocytes were observed until 10 DPC. Antibody-secreting cell probes (ASC-probes) generated from the ALN showed highest reactivity against larval antigens at 5 DPC. This reactivity was predominantly directed against regions between 90 and 100 kDa and 30-35 kDa in the L3 preparation and lower molecular weight antigens in the L4. No reactivity was observed against the adult extract. The 30-35 kDa antigen seemed to exist as a high molecular weight complex in L3 homogenate and was not susceptible to protease K treatment, suggesting it may be non-protein in nature. PMID- 14521581 TI - Cytokine gene expression in dams and foetuses after experimental Neospora caninum infection of heifers at 110 days of gestation. AB - Neospora caninum is a major cause of abortion in cattle. An essential role for Th1 cytokines, such as IFN-gamma and IL-12 in protective immunity against N. caninum in murine models has been indicated. However, little is known about immunity to Neospora in pregnant cattle where a considerable level of immunomodulation may exist. In this study, the immune response of heifers infected early in the second trimester of pregnancy by intravenous inoculation of N. caninum tachyzoites was compared with immune responses in uninfected pregnant heifers. Animals were killed 3 weeks after infection. No abortion was observed in any infected dam, however, transplacental infection was shown to have already taken place. Infection with N. caninum during pregnancy induced significant immune responses in both dams and their foetuses. Infected dams showed significant changes in lymphocyte subpopulations compared with uninfected pregnant animals and these changes were compartmentalized. Increased levels of T lymphocytes were observed in the infected foetuses. Cytokine gene expression analysed by real time RT-PCR showed increased expression of both Th1 and Th2 cytokines in N. caninum infected animals. This cytokine expression could have a role in the transplacental transmission of the parasite and/or mediate tissue damage. PMID- 14521582 TI - Lafoensia pacari extract inhibits IL-5 production in toxocariasis. AB - Toxocariasis is an infection induced by Toxocara canis, an intestinal parasite of dogs. In this study, an experimental murine model of toxocariasis was used to evaluate the anti-inflammatory activity of an ethanolic extract of Lafoensia pacari stem bark. Mice infected with T. canis were treated with L. pacari extract (200 mg/kg, p.o.). Subsequently, we observed a reduction in the number of eosinophils in the peritoneal cavity, bronchoalveolar fluid, blood and bone marrow. Production of interleukin (IL)-5, a major cytokine involved in eosinophilic differentiation, proliferation and activation, is also an important marker for infection. The reduced levels of IL-5 observed in serum, lung homogenates and bronchoalveolar fluid demonstrated the anti-inflammatory mechanisms of L. pacari. Larvae recovery from infected mice treated with L. pacari was comparable with that from untreated mice, suggesting that L. pacari is not toxic to the parasite. Nonetheless, our results demonstrate a potential therapeutic effect of L. pacari extract in IL-5-mediated inflammatory diseases and provide new prospects for the development of drugs to treat IL-5-dependent allergic diseases such as parasite infection and asthma. PMID- 14521583 TI - AIDS, hepatitis and hemophilia in the 1980s: memoirs from an insider. PMID- 14521584 TI - Antithrombotic prophylaxis for women with thrombophilia and pregnancy complications--Yes. PMID- 14521585 TI - Antithrombotic prophylaxis for women with thrombophilia and pregnancy complications--No. PMID- 14521586 TI - TAFI and wound healing: closing a knowledge gap. PMID- 14521587 TI - Thrombin's faces revealed: cellular effects include induction of neoangiogenesis. PMID- 14521588 TI - Roles of factor XI, platelets and tissue factor-initiated blood coagulation. PMID- 14521589 TI - Impaired healing of cutaneous wounds and colonic anastomoses in mice lacking thrombin-activatable fibrinolysis inhibitor. AB - Plasmin and other components of the plasminogen activation system play an important role in tissue repair by regulating extracellular matrix remodeling, including fibrin degradation. Thrombin-activatable fibrinolysis inhibitor (TAFI) is a procarboxypeptidase that, after activation, can attenuate plasmin-mediated fibrin degradation by removing the C-terminal lysine residues from fibrin, which play a role in the binding and activation of plasminogen. To test the hypothesis that TAFI is an important determinant in the control of tissue repair, we investigated the effect of TAFI deficiency on the healing of cutaneous wounds and colonic anastomoses. Histological examination revealed inappropriate organization of skin wound closure in the TAFI knockout mice, including an altered pattern of epithelial migration. The time required to completely heal the cutaneous wounds was slightly delayed in TAFI-deficient mice. Healing of colonic anastomoses was also impaired, as reflected by decreased strength of the tissue at the site of the suture, and by bleeding complications in 3 of 14 animals. Together, these abnormalities resulted in increased mortality in TAFI-deficient mice after colonic anastomoses. Although our study shows that tissue repair, including re epithelialization and scar formation, occurs in TAFI-deficient mice, TAFI appears to be important for appropriate organization of the healing process. PMID- 14521590 TI - Thrombin induces neoangiogenesis in the chick chorioallantoic membrane. AB - Most tumors have constitutively active tissue factor on their surface, capable of generating thrombin in the surrounding environment, and thrombosis is associated with cancer. Thrombin is known to induce a malignant phenotype by enhancing tissue adhesion and cell growth in vitro and in vivo in mice. Because tumors require angiogenesis for growth, we examined whether thrombin induces neoangiogenesis in a physiologically intact in vivo model. Thrombin (0.1 U mL-1) induced neoangiogenesis in the chick chorioallantoic membrane over a 24-72-h period by approximately 2-3-fold. This was inhibited by the potent thrombin inhibitor, hirudin and shown to have its mode of action by ligation of the thrombin protease-activated receptor, PAR-1. The thrombin receptor activation peptide, SFLLRNPNDKYEPF (200 microm) also enhanced neoangiogenesis c. 2-3-fold. Thrombin-induced neoangiogenesis was accompanied by the induction of vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2) mRNA at 24-48 h (approximately 2-fold) as determined by semi-quantitative reverse transcriptase polymerase chain reaction. Thrombin-induced neoangiogenesis was inhibited to baseline level by the specific angiogenesis receptor inhibitors KDR-Fc (vs. VEGF) and Tie-2-Fc (vs. Ang-1 and Ang-2), as well as the non-specific angiogenesis inhibitor thrombospondin-1. Thrombin-induced neoangiogenesis was also inhibited to baseline level by agents known to inhibit thrombin receptor signaling in other cells: G-coupled protein receptor inhibitor, pertussis toxin (40 pg per egg), protein kinase C inhibitor, bisindolylmaleimide (1 microm per egg), MAP kinase inhibitor, PD980598 (10 microm per egg) and PI3 kinase inhibitor, LY294002 (0.25 microm per egg). Thus angiogenesis is stimulated by thrombosis, which could help explain the enhancement of experimental tumorigenesis by thrombin. PMID- 14521591 TI - The function of factor XI in tissue factor-initiated thrombin generation. AB - The influence of plasma and platelet factor (F)XI on thrombin generation initiated with 10 pm tissue factor (TF) in a synthetic coagulation model was evaluated in the presence of either 2 x 108 mL-1 platelets or the equivalent (2 microm) phospholipids. In either system, with all proteins present at physiological concentrations, FXI (30 nm) had no effect on thrombin generation. With phospholipids in the absence of FXI, an increase in vitamin K-dependent proteins (VKDP) (up to 500%) significantly prolonged the initiation phase of thrombin generation and decreased maximum thrombin levels. The inhibition was principally caused by the elevated prothrombin and FIX concentrations. When 30 nm FXI was added with elevated VKDP and phospholipids, the initiation phase was decreased and the maximum thrombin levels generated substantially increased. In experiments with platelets (with and without plasma FXI), an increase in VKDP had little effect on the initiation phase of thrombin generation. These data indicate that (i) FXI has no effect on thrombin generation at 10 pm TF and physiological concentrations of VKDP; (ii) platelets and plasma FXI are able to compensate for the inhibitory effects of elevated VKDP. PMID- 14521592 TI - Nitric oxide in vascular biology. AB - Nitric oxide is a highly versatile heterodiatomic molecule that effects a variety of actions in the vasculture. Originally identified as a principal determination of vascular tone, nitric oxide has since been recognized to exert anti thrombotic, antiproliferative, and anti-inflammatory effects in the vasculture. At higher concentrations and in the setting of other oxidants, nitric oxide can promote vascular pathology. In this review, we summarize the molecular mechanisms of nitric oxides actions in vascular biology and pathology. PMID- 14521593 TI - Comparison of ximelagatran, an oral direct thrombin inhibitor, with enoxaparin for the prevention of venous thromboembolism following total hip replacement. A randomized, double-blind study. AB - BACKGROUND: Prophylaxis is recommended following total joint replacement because of the high risk of venous thromboembolism (VTE). Postoperative low-molecular weight heparin (LMWH) reduces the incidence of venographically detected deep vein thrombosis (DVT) to about 10-15% in total hip replacement (THR) patients. Ximelagatran is a novel, oral direct thrombin inhibitor that selectively and competitively inhibits both free and clot-bound thrombin. We compared the efficacy and safety of ximelagatran with those of enoxaparin for the prevention of VTE in patients undergoing THR. METHODS: This was a prospective, randomized, multicenter, double-blind study conducted principally in the USA and Canada. Patients received fixed-dose oral ximelagatran 24 mg bid or subcutaneous enoxaparin 30 mg bid and matched placebo for 7-12 days; both regimens were initiated the morning after surgery. The incidence of VTE (by postoperative day 12) included thrombosis determined by mandatory venography of the leg on which surgery was performed and symptomatic, objectively proven DVT or pulmonary embolism (PE). VTE and bleeding events were interpreted by an independent central adjudication committee for primary analysis. RESULTS: Of the 1838 patients randomized, 1557 had either adequate venography or symptomatic, proven VTE (efficacy population). Overall rate of venography acceptable for evaluation was 85.4%. Overall rates of total VTE were 7.9% (62 of 782 patients) in the ximelagatran group and 4.6% (36 of 775 patients) in the enoxaparin group, with an absolute difference of 3.3% and a 95% confidence interval for the difference of 0.9% to 5.7%. Proximal DVT and/or PE occurred in 3.6% (28 of 782 patients) in the ximelagatran group and 1.2% (nine of 774 patients) in the enoxaparin group. Major bleeding events were observed in 0.8% (seven of 906) of the ximelagatran-treated patients and in 0.9% (eight of 910) of the enoxaparin-treated patients (P > 0.95). Non-inferiority of ximelagatran 24 mg bid based on a prespecified margin of 5% was not met, resulting in superiority of the enoxaparin regimen. CONCLUSIONS: Both ximelagatran and enoxaparin decreased the overall rate of VTE compared with that reported historically. However, in this study, enoxaparin 30 mg bid was more effective than ximelagatran 24 mg bid for prevention of VTE in THR. Oral ximelagatran was used without coagulation monitoring, was well tolerated, and had bleeding rates comparable to those of enoxaparin. Further refinement by testing a higher dose of ximelagatran in the patients undergoing THR is warranted. PMID- 14521594 TI - Is mild normobaric hypoxia a risk factor for venous thromboembolism? AB - BACKGROUND: Modern air travel entails a cabin altitude between 1520 and 2440 m (5000-8000 ft) and thus exposure to mild hypoxia. There is debate as to whether hypoxia is causally related to venous thromboembolism (VTE) occurring during or after travel. One study suggested that a short period of hypobaric hypoxia causes activation of coagulation. OBJECTIVES: To test the hypothesis that hypoxia alone (normobaric hypoxia) causes activation of coagulation, possibly through endothelial cell activation. METHODS: Six healthy male volunteers were exposed for 3 h, while seated, on two separate occasions to (i) dry air (control) and (ii) hypoxic gas mixture (12.8% O2 in N2, equivalent to breathing air at 3660 m [12000 ft]). RESULTS: There were no differences in hemostatic or endothelial markers between control and hypoxic groups, but platelet and leukocyte counts increased and were significantly higher in the hypoxic group. There were increases in fibrinogen and von Willebrand factor, as well as rheological changes, but these were not significantly different between control and hypoxic groups. CONCLUSIONS: This small study does not support the previous suggestion that hypoxia causes activation of coagulation, and suggests that immobility induced rheological changes may be more significant in the etiology of VTE occurring during or after travel. PMID- 14521595 TI - Thrombophilic families with inheritably associated high levels of coagulation factors VIII, IX and XI. AB - We describe six families in which associated high levels of coagulation factors (F) XI, FIX and FVIII (each with a plasma concentration higher than the 95th percentile found in a control group of 500 asymptomatic individuals: respectively, 135, 145 and 155 IU dL-1) were inherited as a dominant autosomic genetic traits. In these six families, this syndrome is associated with venous thromboembolic events (Odds ratio 41 [4.9-353], P = 0.0006). It seems to predispose to idiopathic events and, as age increases, is often associated with recurrence. First thrombotic episodes occur in young patients (50% of the carriers are symptomatic at the age of 32 years) and in women, can be unmasked by hormonal treatments, mainly oral contraceptives. The association of high levels of coagulation FXI, FIX and FVIII is thus a new rare high-risk inherited thrombophilia syndrome. PMID- 14521596 TI - Prospective double-blind randomized study of the effects of four intravenous fluids on platelet function and hemostasis in elective hip surgery. AB - A prospective randomized double-blind study was performed to determine the effects of three colloids, Haemaccel, Gelofusine and albumin, and also saline on platelet activation, platelet aggregation (induced by adenosine diphosphate (ADP), epinephrine, collagen) platelet agglutination by ristocetin and other hemostatic variables in 55 patients undergoing primary unilateral total hip replacement. The fluids were administered according to normal clinical practice and assessments were made immediately before, at the end, and 2 h after the end of surgery. Surgery was accompanied by thrombin generation (increases in thrombin/antithrombin III complex, prothrombin F1 +2 fragment) platelet activation (betaTG) and compromised coagulation. Generally, the platelet activation appeared to result in platelet desensitization and brought about a persistent reduction in platelet aggregation to ADP and epinephrine, irrespective of the fluid used. Additionally, Haemaccel and Gelofusine inhibited ristocetin induced platelet agglutination and albumin inhibited collagen-induced platelet aggregation. Gross inhibitory effects of Haemaccel that had been predicted from an earlier in vitro study did not occur. Particular fluids had selective additional effects on the hemostatic system. Albumin infusion served to maintain plasma albumin at normal concentrations postsurgery. The two gelatin preparations, Haemaccel and Gelofusine, maintained plasma viscosity. All three colloids led to a transient increase in activated partial thromboplastin time postsurgery and also a transient fall in the concentration of factor VIII, which were accompanied by a transient increase in bleeding time, but there was no measurable increase in blood loss. Inhibition of platelet aggregation by certain colloids may provide additional protection against the increased thrombotic risk in patients following major surgery. PMID- 14521597 TI - Effect of tamoxifen at low doses on ultrasensitive C-reactive protein in healthy women. AB - The use of tamoxifen as a breast cancer preventive agent may be contraindicated by an increased risk of endometrial cancer and venous thromboembolic events, particularly in postmenopausal women. Since these estrogenic effects may be dose related, a dose reduction may reduce toxicity. We have recently shown a comparable activity of lower doses of tamoxifen on putative surrogate biomarkers of cardiovascular disease and breast cancer. To provide further insight into the effect of tamoxifen at low doses on the cardiovascular system, we compared the effect of three different doses on circulating levels of C-reactive protein (CRP), an independent risk marker for cardiovascular disease (CVD), which was lowered by tamoxifen at the standard dose of 20 mg day-1 in previous studies. We compared the changes in CRP after 2 months of either placebo (n = 24), or tamoxifen 10 mg alternate daily (n = 26), or 10 mg day-1 (n = 22), or 20 mg day-1 (n = 19) in healthy women aged 35-70 years. The median percent change was -2.2% (95% CI, -23.3 to 42.8) with placebo, -39.1 (95% CI, -59.9 to -28.7) with 10 mg alternate daily, -56.9% (95% CI, -68.6 to -38.4) with 10 mg day-1 and -42.9% (95% CI, -62.6 to 1.6) with 20 mg day-1 (P = 0.291 for the linear dose-response trend). Similar results were obtained when the data were classified according to blood tamoxifen concentrations, with a median reduction of 47% (95% CI, 65.5 36.3) for women with low tamoxifen concentrations (< 30 ng mL-1). We conclude that tamoxifen at low doses is able to lower ultrasensitive CRP and that this might be associated with a beneficial effect on CVD. PMID- 14521598 TI - Thrombosis in inherited factor VII deficiency. AB - Thrombosis in congenital factor (F) VII deficiency was investigated through extensive phenotypic and molecular-genetic studies. Patients with a history of thrombosis among 514 entries in the FVII Deficiency Study Group database were evaluated. Thrombotic events were arterial in one case, disseminated intravascular coagulation in another and venous in seven. Gene mutations were characterized in eight patients: three were homozygous, three compound heterozygous and two heterozygous. FXa and IIa generation assays were consistent with the genetic lesions. One patient was heterozygous for the FV Leiden and one for the FIIG20210A mutation. In seven patients, surgical interventions and/or replacement therapies had a close temporal relationship with thrombosis, while in the remaining, events were apparently spontaneous. Thromboses were not associated with any specific age, phenotype, mutation zygosity or thrombophilic abnormalities. In particular, severe FVII deficiency did not seem to offer protection from strong thrombosis risk factors such as surgery and replacement therapy. PMID- 14521599 TI - Recognition of coagulation factor VIII by CD4+ T cells of healthy humans. AB - Hemophilia A patients treated with coagulation factor (F)VIII may develop an anti FVIII immune response. Anti-FVIII antibodies may occur also in healthy subjects. To understand the extent to which an immune response to FVIII occurs in healthy subjects, we investigated the proliferative response of blood CD4+ T cells from 90 blood donors to FVIII and to pools of overlapping synthetic peptides spanning the sequences of individual FVIII domains (A1-A3, C1-C2). Most subjects responded to FVIII and several FVIII domains. Men had stronger responses to FVIII than women, and older subjects than younger subjects. The domain-induced responses were weaker than the FVIII-induced responses, yet their intensity in individual subjects correlated with that of the response to FVIII. We examined whether Th1 and/or Th2 cells responded to FVIII in 68 subjects, by determining the CD4+ T cells that secreted interferon-gamma (IFN-gamma) or interleukin (IL)-5 after stimulation with FVIII: 25 subjects had FVIII-specific IFN-gamma-secreting cells, and seven of them had also FVIII-specific IL-5-secreting cells. None had only IL 5-secreting cells. Thus, a CD4+ T cell response to FVIII, which first involves Th1 cells, is common among subjects with a normal procoagulant function. PMID- 14521600 TI - The cost-effectiveness of fondaparinux compared with enoxaparin as prophylaxis against thromboembolism following major orthopedic surgery. AB - BACKGROUND: The selective antithrombotic fondaparinux is more effective than the low-molecular-weight heparin enoxaparin for prevention of venous thromboembolism (deep-vein thrombosis [DVT] or pulmonary embolism) in patients undergoing major orthopedic surgery, but its cost-effectiveness is undetermined. OBJECTIVES: To evaluate the cost-effectiveness of fondaparinux relative to enoxaparin as prophylaxis against venous thromboembolism (VTE) for patients undergoing total hip replacement, total knee replacement or hip fracture surgery in the UK. PATIENTS/METHODS: A decision analysis model was created simulating the impact of fondaparinux and enoxaparin on patient outcomes and costs over various time points up to 5 years following surgery. The main outcome measures were treatment costs per patient and the incidence of clinical VTE and VTE-related deaths. A weighted (combined) cohort reflects the proportion of patients undergoing these procedures in 2000/2001. RESULTS: In the combined cohort, compared with enoxaparin, fondaparinux is expected to produce 20 fewer clinical VTE events and 3.2 fewer VTE-related deaths per 1000 procedures at 5 years. Cost savings at 5 years are pound 27 per patient with fondaparinux (discounted at 6% per year). In each of the three surgical groups, fondaparinux leads to lower expected costs per patient and to a smaller number of VTE events and VTE-related deaths. RESULTS are sensitive to the price difference between fondaparinux and enoxaparin and variation in the rate of late DVT. The analysis is robust to variations in all other key parameters. CONCLUSIONS: Compared with enoxaparin, fondaparinux is more effective and reduces costs to the healthcare system. At current prices, fondaparinux is the recommended strategy in the UK for prophylaxis following major orthopedic surgery. PMID- 14521601 TI - Enhanced in vitro procoagulant and antifibrinolytic potential of superactive variants of recombinant factor VIIa in severe hemophilia A. AB - BACKGROUND: Recombinant coagulation factor VIIa (rFVIIa) is generally accepted for treatment of patients with inhibitor-complicated hemophilia. Recently, rFVIIa variants with a specific enhancement of the tissue factor (TF)-independent proteolytic activity have been described. OBJECTIVES: The procoagulant and [thrombin-activatable fibrinolysis inhibitor (TAFI)-dependent] antifibrinolytic potentials of two superactive rFVIIa variants were compared with those of wild type rFVIIa in a hemophilic setting. PATIENTS AND METHODS: Clot lysis assays were performed in plasma from six patients with inhibitor-complicated hemophilia A or in antibody-induced factor VIII-deficient platelet-rich plasma in the presence of different concentrations of the rFVIIa variants. RESULTS AND DISCUSSION: In the plasma model, M298Q-rFVIIa had a moderately increased procoagulant and antifibrinolytic potential, whereas V158D/E296V/M298Q/K337A-rFVIIa had a strongly increased procoagulant and antifibrinolytic activity compared with wild-type rFVIIa. The increased antifibrinolytic potential of the rFVIIa variants was completely dependent on enhancement of TAFI activation. In the platelet-rich plasma model similar results were obtained. The presence of TF was mandatory for clot formation in the absence of exogenous rFVIIa. At lower concentrations of rFVIIa (wild-type or variants), clot formation did occur but was significantly slower when TF activity was blocked. At increasing concentrations of rFVIIa, clotting times were no longer dependent on TF. In conclusion, should a TF independent mechanism be involved in the efficacy of rFVIIa in patients with hemophilia, the superactive rFVIIa variants studied here might be clinically advantageous, as both procoagulant and antifibrinolytic potencies are significantly enhanced compared with those of wild-type rFVIIa. This ought to result in more efficient cessation of bleeding episodes and reduced risk of rebleeding. PMID- 14521602 TI - A functional haplotype in the 5' flanking region of the factor VII gene is associated with an increased risk of coronary heart disease. AB - AIMS: The aim of this study was to investigate associations between coronary heart disease risk and polymorphisms in the coagulation factor (F)VII gene in participants of a large prospective study. METHODS: One thousand nine hundred and fifty-seven men were genotyped for four FVII polymorphisms, -670A-->C, -402G-->A, a 10 base pair insertion at -323 (0 > 10) in the promoter, and R353Q in the structural gene. Associations among genotypes and estimated haplotypes, plasma FVII levels, and coronary heart disease risk were evaluated, and the function of the promoter polymorphisms was assessed in reporter gene assays. RESULTS: The 670A-->C and -402G-->A polymorphisms were in complete allelic association. The haplotype containing -670C and -402A (frequency =0.23) was associated with significantly increased plasma FVII coagulant activity and increased risk of an initial coronary event, particularly acute myocardial infarction, which remained after correction for conventional risk factors. In contrast, the -323 insertion and Q353 alleles (frequency =0.11 and 0.10, respectively) were associated with decreased plasma FVII levels, but hazard ratios for coronary events in carriers of these alleles were not significantly different from unity. In transiently transfected hepatoma cells, increased basal expression of the reporter gene was directed by a promoter fragment with rare haplotype -670C/-630G/-402A rather than by a promoter fragment with common haplotype -670A/-630A/-402G; -402A was not responsible for this effect. CONCLUSIONS: The promoter haplotype, -670C/ 630G/402A, was associated with significantly increased plasma FVII coagulant activity, risk of an initial coronary event, particularly acute myocardial infarction, and reporter gene expression. PMID- 14521603 TI - Interaction between oral contraceptive use and coagulation factor levels in deep venous thrombosis. AB - Deep venous thrombosis is a multicausal disease, i.e. more than one risk factor needs to be present to cause the disease. Oral contraceptive use increases the risk of venous thrombosis but since not all women using oral contraceptives develop thrombosis, the presence of additional risk factors in patients is likely. The aim of this study was to assess the joint effect of oral contraceptive use and the levels of procoagulant factors (F)(FII, FV, FVII, FVIII, FIX, FX, FXI, FXII, FXIII and fibrinogen). Data of premenopausal women were re-analyzed in the Leiden Thrombophilia Study. The highest relative risks were observed for the combination of oral contraceptive use and high levels (>90th percentile) of FII (Odds Ratio [OR]OC+FII 10.1; 95% confidence interval [CI] 3.5-29.0), FV (OROC+FV 12.6; 95% CI 3.8-41.5), and FXI (OROC+FXI 11.9; 95% CI 3.6-39.2) and low levels (< 10th percentile) of FXII (OROC+FXII 12.3; 95% CI 2.4-63.0). No interaction was observed between oral contraceptive use and high levels of the other coagulation factors, i.e. the joint effect of these risk factors did not exceed the sum of the separate effects. The results of this study indicate that the risk for the joint effects of oral contraceptive use and coagulation factor levels are minor compared with the joint effect of oral contraceptive use and the FV Leiden mutation (RR > 30). PMID- 14521604 TI - Measurement of von Willebrand factor activity: relative effects of ABO blood type and race. AB - Tests based on three different principles are reported to measure the activity of von Willebrand factor (VWF): ristocetin cofactor (VWF:RCo), collagen binding (VWF:CB), and the so-called "activity ELISA" (VWF:MoAb). We measured these and other diagnostic parameters in a population of 123 randomly selected female study controls, age 18-45 years. Type O subjects had significantly lower levels than non-O subjects in each test. Race differences were seen in all tests except VWF:RCo, with Caucasians having significantly lower levels than African Americans. ABO differences accounted for 19% of the total variance in VWF:Ag (P < 0.0001) and race for 7% (P < 0.0001), for a total of 26%. Both effects were mediated through VWF:Ag and were independent. VWF:Ag level was the primary determinant of VWF function, accounting for approximately 60% of the variance in VWF:RCo and VWF:CB and 54% of the variance in factor VIII. The ratio VWF:RCo/VWF:Ag differed significantly by race within blood group. The median ratios were 0.97 for type O Caucasians vs. 0.79 for type O African-Americans and 0.94 for non-O Caucasians vs. 0.76 for non-O African-Americans. The ratio VWF:CB/VWF:Ag did not vary. This suggests racial differences in the interaction of VWF with GP1b but not with subendothelium. Alternatively, VWF:RCo may be regulated to maintain a relatively constant plasma level in the presence of excessive VWF:Ag. This heterogeneity within the normal population is partially responsible for the difficulty in defining diagnostic limits for von Willebrand disease. PMID- 14521605 TI - Identification of a novel point mutation in platelet glycoprotein Ibalpha, Gly to Ser at residue 233, in a Japanese family with platelet-type von Willebrand disease. AB - BACKGROUND: Interaction between platelet glycoprotein (GP)Ibalpha and von Willebrand factor (VWF) has critical roles in both physiological hemostasis and thrombosis. Platelet-type von Willebrand disease (plt-VWD) is a congenital bleeding disorder characterized by gain-of-function mutations of GPIbalpha. To date, two mutations in GPIbalpha, G233V and M239V, have been reported in four unrelated families with plt-VWD. OBJECTIVE: The present study aimed to determine whether G233S of GPIbalpha, a new mutation observed in plt-VWD patients, causes the plt-VWD phenotype and to examine whether conversions to other residues at this position affect VWF binding. PATIENTS AND METHODS: The propositus was a 3 year-old Japanese male. He displayed bleeding symptoms and moderate thrombocytopenia. His brother was similarly affected. Platelets from both patients were analyzed by ristocetin- or shear-induced platelet aggregation. DNA sequencing was performed to analyze the GPIbalpha sequence. We examined the 125I labeled VWF binding using a series of recombinant GPIbalpha fragments with different residues at position 233 (G233S, G233A, G233K, and G233D) together with naturally occurring mutations previously reported in patients (G233V and M239V). RESULTS: Platelet function analysis indicated that platelets from both patients had a typical plt-VWD phenotype. DNA sequencing analysis showed a heterozygous mutation of Gly to Ser at residue 233 of GPIbalpha in both patients. The 125I labeled VWF binding to mutant compared with the wild type displayed three patterns, gain-of-function (G233S, G233V, and M239V), equivalent function (G233A), and loss-of-function (G233K and G233D). CONCLUSIONS: The G233S is a molecular basis of plt-VWD, and residue 233 plays critical roles in regulating VWF binding. PMID- 14521606 TI - Redistribution of glycoprotein Ib within platelets in response to protease activated receptors 1 and 4: roles of cytoskeleton and calcium. AB - Thrombin activates human platelets by hydrolyzing the protease-activated receptors PAR-1 and PAR-4, exposing new N-terminal sequences which act as tethered ligands, and binding to glycoprotein (GP) Ib, whose surface accessibility transiently decreases when platelets are stimulated by the enzyme. In an attempt to better understand this latter process, we used the peptides SFLLRNPNDKYEPF (PAR-1-AP or TRAP) and AYPGKF (PAR-4-AP) to study whether hydrolysis of both PAR receptors leads to GPIb redistribution. Both peptides induced surface clearance of GPIb with a maximum at 2 min and 5 min for PAR-1-AP and PAR-4-AP, respectively, followed by a slow return to the surface with levels normalizing between 30 and 60 min. Translocation was associated with the formation of clusters of GPIb as revealed by fluorescence microscopy. This transient redistribution of GPIb was blocked by cytochalasin D and in large part by the membrane permeable Ca2+ chelator, BAPTA. The inhibitor of phosphatidylinositol 3-kinase and myosin light chain kinase, wortmannin, did not significantly modify internalization of GPIb, although its return to the surface was delayed for PAR-1-AP. PAR receptor-mediated association of GPIb to the insoluble cytoskeleton was blocked by cytochalasin D, while BAPTA alone increased and stabilized the presence of GPIb. Globally, immunoprecipitation experiments and analysis of the cytoskeleton confirmed that GPIb translocation is powered by a contractile mechanism involving Ca2+ mobilization, actin polymerization, and myosin incorporation into the cytoskeleton and that both PAR-1 and PAR-4 can activate this process. PMID- 14521607 TI - Involvement of the beta3 E749ATSTFTN756 region in stabilizing integrin alphaIIbbeta3-ligand interaction. AB - Platelet integrin alphaIIbbeta3 must be activated via intracellular mechanisms before it binds soluble ligands, and it is thought to be activated at its extracellular site by surface-bound ligands. Integrin activation is associated with rearrangement of the cytoskeleton and phosphorylation of proteins that become localized in focal contacts. In these processes, the cytoplasmic tail of the beta-subunit plays a central role. We introduced peptides homologous to the E749ATSTFTN756 domain (E-N peptide) and the T755NITYRGT762 domain (T-T peptide) of beta3 in streptolysin O-permeabilized platelets and analyzed the initial interaction with soluble fibronectin, fibrinogen and PAC-1 after stimulation with thrombin. E-N peptide left the initial binding of fibronectin intact but interfered with stable receptor occupancy. E-N peptide also inhibited fibrinogen binding, thereby reducing the formation of large aggregates. Strikingly, E-N peptide did not disturb the binding of PAC-1, which is known to reflect activation of the integrin. E-N peptide also inhibited tyrosine phosphorylation of focal adhesion kinase, a response known to be dependent on alphaIIbbeta3. T-T peptide did not affect these processes. In a model for outside-in integrin activation, E-N peptide disrupted the binding of CHO cells expressing alphaIIbbeta3 to surface-bound ligand. Again, T-T peptide had no effect. We conclude that the E749ATSTFTN756 region of the beta3-tail stabilizes the binding of soluble and surface-bound ligand to integrin alphaIIbbeta3 via a mechanism that involves the phosphorylation of FAK. PMID- 14521608 TI - Aspirin and salicylate inhibit colon cancer medium- and VEGF-induced endothelial tube formation: correlation with suppression of cyclooxygenase-2 expression. AB - To determine whether aspirin and salicylate suppress colon cancer cell-mediated angiogenesis, we evaluated the effects of aspirin and sodium salicylate on endothelial tube formation on Matrigel. Aspirin and sodium salicylate concentration-dependently inhibited human endothelial cell (EC) tube formation induced by conditioned medium collected from DLD-1, HT-29 or HCT-116 colon cancer cells. Aspirin and sodium salicylate at pharmacological concentrations were equally effective in blocking tube formation. Neutralizing antivascular endothelial growth factor (VEGF) antibodies blocked colon cancer medium-induced tube formation. VEGF receptor 2 but not receptor 1 antibodies inhibited tube formation to a similar extent as anti-VEGF antibodies. These results indicate that VEGF interaction with VEGF receptor 2 is the primary mechanism underlying colon cancer-induced angiogenesis. Aspirin or sodium salicylate inhibited VEGF induced tube formation in a concentration-dependent manner comparable to that of inhibition of colon cancer medium-induced endothelial tube formation. It has been shown that cyclooxygenase-2 (COX-2) is pivotal in cancer angiogenesis. We found that colon cancer medium-induced COX-2 protein expression in EC and aspirin or sodium salicylate suppressed the cancer-induced COX-2 protein levels at concentrations correlated with those that suppressed endothelial tube formation. Furthermore, aspirin and sodium salicylate inhibited COX-2 expression stimulated by VEGF. These findings indicate that aspirin and other salicylate drugs at pharmacological concentrations inhibit colon cancer-induced angiogenesis which is correlated with COX-2 suppression. PMID- 14521609 TI - Intimal preatherosclerotic thickening of the coronary arteries in human fetuses of smoker mothers. AB - BACKGROUND: Many studies have described the development of preatherosclerotic coronary artery lesions in infancy. The observations reported in the literature regarding the fetal origin of coronary artery lesions are rare and controversial. OBJECTIVES: To identify the features of preatherosclerotic coronary artery lesions in late fetal stillborns and the possible atherogenic role of maternal cigarette smoking. METHODS; We examined 22 stillborns (13 males and nine females), all of whom had died sine causa after the 32nd week of gestation. All underwent autopsy. Twelve of the mothers smoked over five cigarettes per day before and during the pregnancy. The four major epicardial coronary arteries were isolated along their whole length, embedded in paraffin and serially cut for histologic examination and immunohistochemical studies, particularly searching for the proliferating cell nuclear antigen and c-Fos expression. Alterations of chromosome 7 were also investigated by the fluorescence in situ hybridization technique. RESULTS: In over 50% of the fetuses, almost all from smoker mothers, multifocal structural alterations of coronary walls were evident. The smooth muscle cells (SMCs) presented loss of polarity, forming columns perpendicular to the axis of the media and infiltrating the subendothelial connective tissue. Increased amounts of mucoid ground substance were also observed in the subendothelial connective tissue. In all the cases with coronary alterations, study of the biological markers showed intense c-Fos positivity of the SMCs. CONCLUSIONS: Preatherosclerotic intimal alterations of the coronary arteries are already detectable in the prenatal period and are significantly associated with maternal cigarette smoking. PMID- 14521610 TI - Screening of high factor VIII levels is not recommended in patients with recently diagnosed pulmonary embolism. PMID- 14521611 TI - Availability of technology to evaluate for pulmonary embolism in academic emergency departments in the United States. PMID- 14521612 TI - Prevalence of inherited bleeding disorders in cases of idiopathic menorrhagia: a case of five blind men describing an elephant. PMID- 14521613 TI - Very high TAFI antigen levels are associated with a lower risk of hard coronary events: the PRIME Study. PMID- 14521615 TI - A common mutation in 5,10-methylenetetrahydrofolate reductase (MTHFR) gene in two Arab communities. PMID- 14521614 TI - Role of the fibrinogen gamma-chain sequence gamma316-322 in platelet-mediated clot retraction. PMID- 14521616 TI - Evidence of platelet activation due to tirofiban-dependent platelet antibodies: double trouble. PMID- 14521617 TI - Antibodies to tissue factor pathway inhibitor are uncommonly detected in patients with infection-related antiphospholipid antibodies. PMID- 14521618 TI - Warfarin and acenocoumarol dose requirements according to CYP2C9 genotyping in North-Italian patients. PMID- 14521619 TI - Prothrombin G20210A is not prevalent in North India. PMID- 14521620 TI - A different view of "Toc", as I knew him. PMID- 14521621 TI - Milder bleeding tendency in Glanzmann's thrombasthenia patients inheriting HPA-1b in the homozygous state. PMID- 14521622 TI - Rebuttal to: LMWH vs. LMWH: superior, equivalent or non-inferior? PMID- 14521626 TI - Digging deep--structure-function relationships in the melatonin receptor family. AB - The melatonin receptor family is a small group of receptors within the G protein coupled receptor (GPCR) superfamily. The group comprises of three subtypes which bind melatonin and one member, the melatonin related receptor (MRR), that shares >40% sequence identity with the other melatonin receptors but does not bind melatonin. Identification of two subtypes expressed in the mouse suprachiasmatic nucleus, one of which (MT1) inhibits neuronal firing and the other (MT2) mediating the phase advancing properties of melatonin has given renewed interest to the development of subtype specific compounds for each of the mammalian melatonin receptors. Towards this goal site-directed and chimaeric receptor mutagenesis studies have been performed which have provided some insight into the structure-function relationships of the melatonin receptors. Furthermore, these studies may lead to the identification of the ligand for the orphan MRR. PMID- 14521627 TI - Melatonin prevents apoptosis and enhances HSP27 mRNA expression induced by heat shock in HL-60 cells: possible involvement of the MT2 receptor. AB - Previous studies have reported that melatonin protects cells and tissues against stressful stimuli. In the present study using HL-60 cells, we show that cells acquire increased resistance to apoptosis normally induced by heat shock when they are incubated with melatonin. This effect of melatonin is saturable at nanomolar concentrations and appears to be mediated by the MT2 subtype melatonin receptor. The high affinity melatonin receptor agonist, 2-iodomelatonin, reproduced the melatonin effect while it was fully blocked by the selective MT2 antagonist 4-phenyl-2-propionamidotetraline. The melatonin response to heat shock induced apoptosis was pertussis toxin sensitive and, interestingly, the non selective MT1/MT2 melatonin receptor ligand luzindole was found to display agonistic activity. Furthermore, we provide evidence that melatonin enhanced HSP27 mRNA expression as a result of heat shock - HSP27, is known to play an important role in the defense of cells against apoptosis induced by stressful agents. Together, these results demonstrate that melatonin, likely via receptor mechanisms, interferes with the apoptotic pathway activated by heat shock. PMID- 14521628 TI - Relationship between salivary melatonin levels and periodontal status in diabetic patients. AB - Among other functions, melatonin exerts both antioxidative and immunoregulatory roles. The indoleamine is secreted in the saliva, although its role into the mouth is not known. Diabetic patients frequently display oral cavity pathologies such as periodontal disease (PD), an inflammatory disease coursing with an increase in free radical production. Thus, we compared the degree of PD and interleukin-2 (IL-2) levels with melatonin concentrations in plasma and saliva of diabetic patients. A total of 43 diabetic patients (20 with type I and 23 with type II diabetes) and 20 age- and sex-matched controls were studied. Dental and medical history of all patients was in accordance with the criteria of the WHO. The periodontal status was evaluated by the Community Periodontal Index (CPI). Plasma and salivary melatonin levels were determined by specific commercial radioimmunoassays, and plasma IL-2 was measured using a commercial enzyme-linked immunosorbent assay kit. Diabetic patients had plasma and saliva melatonin levels of 8.98 +/- 7.14 and 2.70 +/- 2.04 pg/mL, respectively. These values were significantly lower (P < 0.001) than those obtained in plasma and saliva of controls (14.91 +/- 4.75 and 4.35 +/- 0.98 pg/mL, respectively). Plasma and salivary melatonin concentrations show a biphasic response in diabetic patients. Melatonin decreased in patients with a CPI index of 2, and then increased reaching highest levels in patients with a CPI index of 4. By contrast, IL-2 levels decreased from CPI index 1 to 4. The results indicate that, in diabetic patients, the presence of a marked impairment of the oral status, as assessed by the CPI index, is accompanied by an increase in plasma and salivary melatonin. The increase in salivary melatonin excretion may have a periodontal protective role. PMID- 14521629 TI - N1-acetyl-N2-formyl-5-methoxykynuramine is a product of melatonin oxidation in rats. AB - The product of melatonin oxidation, N1-acetyl-N2-formyl-5-methoxykynuramine (AFMK), was synthesized and a method for its determination in biological samples was developed. High performance liquid chromatography (HPLC) with fluorescence detection provided good sensitivity and selectivity. Wavelengths of 350 and 480 nm were used for excitation and emission, respectively. Serum and retinal homogenates were extracted with chloroform prior to analysis by HPLC. Endogenous AFMK was detected in the retina of rats but the serum concentration of this melatonin metabolite was below the detection limit of the method for measurement. Retinal AFMK concentration was higher during the dark phase of the light/dark cycle, when the retinal melatonin content is maximal. Intraperitoneal administration of melatonin significantly increased serum and retinal AFMK levels. Formation of AFMK from melatonin was also confirmed by in vivo microdialysis with the probe implanted into the brain lateral ventricle. The study shows that AFMK is indeed a product of melatonin oxidation in vivo. The possible physiological significance of melatonin oxidation metabolic pathway is discussed. PMID- 14521630 TI - Effect of pinealectomy on plasma levels of insulin and leptin and on hepatic lipids in type 2 diabetic rats. AB - We previously reported that pharmacological melatonin administration to type 2 diabetic rats reduces hyperinsulinemia and improves the altered fatty-acid metabolism. To determine whether melatonin deficiency exacerbates diabetes associated conditions, we investigated the effect of pinealectomy (i.e. melatonin deficiency) on plasma hormone levels and lipid metabolism in type 2 diabetic Otsuka Long-Evans Tokushima Fatty (OLETF) rats. We compared levels of insulin and leptin, and hepatic lipids in pinealectomized OLETF (PO) rats, sham-operated OLETF (SO) rats and sham-operated healthy Long-Evans Tokushima Otsuka (LETO) (SL) rats 16 and 30 wk after the operation. Plasma glucose and triglycerides were increased in SO and PO rats 30 wk after operation compared with age-matched SL rats. Pinealectomy caused an increase in free cholesterol among the plasma lipids, as compared with SO rats. Sixteen weeks after pinealectomy, typical hyperinsulinemia was observed in PO rats (3.47-fold increase, P < 0.01) as compared with SL rats, whereas at 30 wk, the plasma levels of insulin in PO and SO rats had decreased and there was no significant difference among the three groups. Hepatic triglycerides were increased (1.54-fold, P < 0.005) in PO rats, compared with SO rats. Hepatic acyl-CoA synthetase (ACS) activity was significantly augmented in PO rats at 30 wk (10%, P < 0.01 versus SO group), while microsomal triglyceride transfer protein (MTP) decreased (-27% versus SO, P < 0.05); thus, the increased ACS activity and decreased MTP might have a role in the accumulation of hepatic triglycerides in PO rats. In summary, pinealectomy causes severe hyperinsulinemia and accumulation of triglycerides in the liver, probably owing to the loss of the nocturnal melatonin surge. PMID- 14521631 TI - Melatonin protects against myocardial doxorubicin toxicity in rats: role of physiological concentrations. AB - Doxorubicin (Dox) is a widely used antineoplastic drug. Oxygen radical-induced injury of membrane lipids is considered to be the most important factor responsible for the development of Dox-induced cardiotoxicity. The pineal secretory product, melatonin, is known to be a potent free radical scavenger and its pharmacological concentrations have been shown to reduce Dox-induced cardiac damage. However, the physiological role of melatonin in the prevention of this damage is unknown. We investigated physiological and pharmacological effects of melatonin on Dox-induced changes in the levels of malondialdehyde (MDA), a lipid peroxidation product, and morphological changes in heart. Rats were pinealectomized (Px) or sham-operated (control) 2 months before the studies. Melatonin was administered [4 mg/kg, intraperitoneally (i.p.)] 1 hr before or 24 hr after the administration of a single dose of Dox (20 mg/kg, i.p.) and continued for 2 days. The levels of MDA Dox was found to be significantly higher in the Px rats (55.9 +/- 0.6 nmol/g tissue) than intact control animals (42.6 +/- 0.4). Dox administration to Px and non-Px rats significantly increased the MDA levels. Pre- and post-treatment with melatonin in both Px and intact rats significantly reduced MDA levels. Morphological changes parallelled the MDA alterations. These findings strongly suggest that both physiological and pharmacological concentrations of melatonin are important in protecting the heart from Dox-induced damage in rats. It would seem valuable to test melatonin in clinical trials for prevention of possible heart damage associated with Dox. PMID- 14521632 TI - Melatonin protects against pro-oxidant enzymes and reduces lipid peroxidation in distinct membranes induced by the hydroxyl and ascorbyl radicals and by peroxynitrite. AB - We have investigated the action of melatonin against lipid peroxidation in membranes including brain homogenates (BH), brain and liver microsomes (MIC), and phosphatidylcholine (PC) liposomes, as well as its effect on the activity of pro oxidant enzymes such as constitutive neuronal nitric oxide synthase (cnNOS), xanthine oxidase (XO) and myeloperoxidase (MPO). The liposomes were reconstituted by a dialysis method, lipid peroxidation was monitored using the thiobarbituric reactive substances (TBARS) method and enzyme activities were measured spectrophotometrically. The ascorbyl and hydroxyl free radicals were generated by the reaction of ascorbic acid + FeSO4 and H2O2 + FeCl2, respectively, and peroxynitrite using a mixture of NaNO2 in an alkaline medium. Melatonin protected against lipid peroxidation induced by distinct reactive oxygen species (ROS) in all membranes tested although with different potency, in the following order BH < MIC < PC. The K0.5 for enzyme inhibition by melatonin was determined for nNOS (2.0 +/- 0.1 mm), for XO (0.8 +/- 0.1 mm) and for MPO (0.063 +/- 0.003 mm), the latter one with high affinity. Melatonin showed a weak effect as a nitrogen monoxide (NO) scavenger in the presence of sodium nitroprusside (NO donor) and low reactivity with 1,1-diphenyl-2-picryl hydrazyl (DPPH). These results demonstrate the antioxidant action of melatonin, principally that related to the activity of pro-oxidant enzymes such as XO and MPO. PMID- 14521633 TI - Melatonin: structural characterization of its non-enzymatic mono-oxygenate metabolite. AB - Oxidation of melatonin by Fenton reagents as well as with hypochlorous acid or oxoferryl hemoglobin has been investigated. Analysis of products by low resolution/mass spectra (MS), high resolution/MS, 1H-nuclear magnetic resonance (NMR), 13C-NMR, correlated spectroscopy (COSY) and heterocorrelated spectroscopy (HETCOR) 2D NMR reveals the formation of a single mono-oxygenated product under all conditions and unequivocally assigns the N-[2-(5-methoxy-2-oxo-2,3-dihydro-1H indol-3-yl)-ethyl]-acetamide structure, which had not been previously considered. PMID- 14521634 TI - Protective effect of indoleamines on in vitro ascorbate-Fe2+ dependent lipid peroxidation of rod outer segment membranes of bovine retina. AB - Rod outer segment membranes (ROS) are highly vulnerable to autooxidation because of their high content of long chain polyunsaturated fatty acids (PUFAs). Melatonin and N-acetylserotonin are indoleamines synthesized in the pineal gland, retina and other tissues. These compounds are free radical scavengers and indirect antioxidants because of their stimulatory effect on antioxidative enzymes. We compared the in vitro protective effect of melatonin and N acetylserotonin on the ascorbate-Fe2+ induced lipid peroxidation of PUFAs located in ROS membranes. This process was measured by chemiluminescence and fatty acid composition of total lipids of ROS. We assayed increasing concentrations of melatonin (0-10 mm) and N-acetylserotonin (0-2 mm). In both cases the total cpm originated from light emission (chemiluminescence) was found to be lower in those membranes incubated in the presence of either melatonin or N-acetylserotonin; this decreased proportional to the concentration of the indole. Thus, 10 mm melatonin and 2 mm N-acetylserotonin produced a reduction of 51 +/- 6 and 100% in the total chemiluminescene (lipid peroxidation), respectively. We also noticed a PUFAs protection: the docosahexaenoic acid content decreased considerably when the membranes were submitted to oxidative damage. This reduction was from 37.6 +/ 2.1% in the native membranes to 6.2 +/- 0.8% in those which were peroxidized. These changes were less pronounced in treated ROS membranes; as an example in the presence of 10 mm melatonin or 2 mm N-acetylserotonin we observed a content preservation of 22:6 n-3 (23.6 +/- 1.2 and 39.1 +/- 1.2% respectively). The concentration of each compound required to inhibit 50% of the lipid peroxidation (IC50) was 9.82 mm for melatonin and 0.43 mm for N-acetylserotonin, respectively. N-acetylserotonin shows a protective effect about 20 times higher than that of melatonin. PMID- 14521635 TI - Perfusion of melatonin into the prefrontal cortex disrupts the circadian rhythm of acetylcholine but not of locomotor activity. AB - Extracellular concentrations of acetylcholine (ACh) in the prefrontal cortex displayed a circadian rhythm, with a maximum increase in the dark phase of the light:dark cycle. The increase in ACh related well to the circadian rhythm of the same rats in which a maximal increase of locomotor activity in the dark phase also was observed. Local perfusion of melatonin (200-500 microm), in a dose dependent manner, disrupted the ACh rhythm in the prefrontal cortex but did not affect the circadian rhythm of locomotor activity. It is suggested that the change in the cholinergic transmission during a circadian period in the prefrontal cortex may be under the control of the biological clock through the action of melatonin; however, the prefrontal cortical ACh cycle seems not to be related to the regulation of locomotor activity. PMID- 14521636 TI - Protective effect of melatonin against fractionated irradiation-induced epiphyseal injury in a weanling rat model. AB - The effects of melatonin, a free-radical scavenger and a general antioxidant, on radiation-induced growth plate injury have not been studied previously. The purpose of this study was to determine the potential benefits of sparing longitudinal bone growth by fractionated radiotherapy alone compared with pretreatment with melatonin that provides differential radioprotection of normal cells. Weanling 4-wk-old (75-100 g) male Sprague-Dawley rats were randomly assigned to one of three groups: Group R received fractionated radiation alone (n = 8); groups M5 (n = 8) and M15 (n = 7) received 5 or 15 mg/kg melatonin prior to fractionated radiation, respectively. The distal femur and proximal tibia in the right leg of each animal were exposed to a therapeutic X-irradiation dose (25 Gy total in three fractions) with the contralateral left leg as the non-irradiated control. Melatonin was administered intraperitoneally to the animals 30 min before radiation exposure. Six weeks after treatment, the rats were killed and the lower limbs disarticulated, skeletonized, radiographed, and bone growth was calculated based on measurement of the bone lengths. Fractionated radiation resulted in a mean percent overall limb growth loss of 41.2 +/- 9.5 and a mean percent overall limb discrepancy of 11.2 +/- 2.2. The administration of 5 or 15 mg/kg melatonin before each of the three fractions of radiotherapy reduced the mean percent overall limb growth loss to 33.9 +/- 5.8 and 32.2 +/- 4.5, respectively, and the mean percent overall limb discrepancy to 9.4 +/- 1.6 and 8.9 +/- 1.1, respectively; these values were significantly different compared with irradiation alone (range: P = 0.01-0.04). When compared with Group R, the growth arrest recovered by 5 or 15 mg/kg melatonin was 19.7 and 24.1% for the tibia, 7 and 18.6% for the femur, and 17.7 and 21.8% for the total limb, respectively. These results support further investigation of melatonin in combination with fractionation for potential use in growing children requiring radiotherapy to the extremity for malignant tumors. PMID- 14521637 TI - Urinary 6-hydroxy-melatonin-sulfate excretion and circadian rhythm in patients with restless legs syndrome. AB - The precise etiology of the restless legs syndrome (RLS) is unknown. Sensory and motor symptoms of RLS worsen during evening/night, coincident with the physiological peak of pineal melatonin excretion. Decreased melatonin levels have been reported in insomnia, which is an associated feature of RLS. Melatonin substitution improved insomnia. A potential association between the idiopathic RLS (iRLS) and alterations in melatonin excretion was therefore explored. Daytime (7:00-22:00 hr) and night-time (22:00-7:00 hr) urinary excretion of 6-OH melatonin-sulfate (aMLTs) was measured in 15 patients with iRLS and 11 controls by a radioimmunoassay. There was no significant difference between daytime and night-time urinary aMLTs excretion in iRLS as compared with controls (daytime: 6.14 +/- 5.20 ng versus 5.02 +/- 5.11 ng, NS; night-time: 21.07 +/- 17.05 ng versus 22.92 +/- 16.52 ng, NS). Our data do not provide evidence for a decrease of cumulative melatonin production in iRLS. Insomnia in RLS does not seem to be correlated with a deficit of melatonin. PMID- 14521638 TI - Randomized controlled trial of education and feedback for implementation of guidelines for acute low back pain. AB - OBJECTIVE: The effect of clinical guidelines on resource utilization for complex conditions with substantial barriers to clinician behavior change has not been well studied. We report the impact of a multifaceted guideline implementation intervention on primary care clinician utilization of radiologic and specialty services for the care of acute low back pain. DESIGN: Physician groups were randomized to receive guideline education and individual feedback, supporting patient education materials, both, or neither. The impact on guideline adherence and resource utilization was evaluated during the 12-month period before and after implementation. PARTICIPANTS: Fourteen physician groups with 120 primary care physician and associate practitioners from 2 group model HMO practices. INTERVENTIONS: Guideline implementation utilized an education/audit/feedback model with local peer opinion leaders. The patient education component included written and videotaped materials on the care of low back pain. MAIN RESULTS: The clinician intervention was associated with an absolute increase in guideline consistent behavior of 5.4% in the intervention group versus a decline of 2.7% in the control group (P =.04). The patient education intervention produced no significant change in guideline-consistent behavior, but was poorly adopted. Patient characteristics including duration of pain, prior history of low back pain, and number of visits during the illness episode were strong predictors of service utilization and guideline-consistent behavior. CONCLUSIONS: Implementation of an education and feedback-supported acute low back pain care guideline for primary care clinicians was associated with an increase in guideline-consistent behavior. Patient education materials did not enhance guideline effectiveness. Implementation barriers could limit the utility of this approach in usual care settings. PMID- 14521639 TI - A randomized controlled trial comparing internet and video to facilitate patient education for men considering the prostate specific antigen test. AB - BACKGROUND: Little is known about the relative advantages of video versus internet-based decision aids to facilitate shared medical decision making. This study compared internet and video patient education modalities for men considering the prostate specific antigen (PSA) test. METHODS: Two hundred and twenty-six men, aged 50 years or older, and scheduled to complete a physical examination at an HMO Health Appraisal Clinic were randomly assigned to access a website (N = 114) or view a 23-minute videotape in the clinic (N = 112) prior to deciding whether they wanted to be screened for prostate cancer. RESULTS: There were no between-groups differences in participants' ratings of convenience, effort, or satisfaction following exposure to the decision aid. Participants assigned to the video group were more likely to review the materials than individuals assigned to the internet group (98.2% vs 53.5%). Participants in the video group showed significantly greater increases in PSA knowledge and were more likely to decline the PSA test than individuals assigned to the internet group. However, participants in the internet group who reviewed the entire online presentation showed similar increases in PSA knowledge as video participants. Only 5% of all participants visited other websites to inform themselves about the PSA test. CONCLUSIONS: Overall, the video was significantly more effective than the Internet in educating participants about benefits and risks of PSA screening. PMID- 14521640 TI - Could we have known? A qualitative analysis of data from women who survived an attempted homicide by an intimate partner. AB - OBJECTIVE: To examine in-depth the lives of women whose partners attempted to kill them, and to identify patterns that may aid in the clinician's ability to predict, prevent, or counsel about femicide or attempted femicide. DESIGN: Qualitative analysis of 30 in-depth interviews. SETTING: Six U.S. cities. PARTICIPANTS: Thirty women, aged 17-54 years, who survived an attempted homicide by an intimate partner. RESULTS: All but 2 of the participants had previously experienced physical violence, controlling behavior, or both from the partner who attempted to kill them. The intensity of the violence, control, and threats varied greatly, as did the number of risk factors measured by the Danger Assessment, defining a wide spectrum of prior abuse. Approximately half (14/30) of the participants did not recognize that their lives were in danger. Women often focused more on relationship problems involving money, alcohol, drugs, possessiveness, or infidelity, than on the risk to themselves from the violence. The majority of the attempts (22/30) happened around the time of a relationship change, but the relationship was often ending because of problems other than violence. CONCLUSIONS: Clinicians should not be falsely reassured by a woman's sense of safety, by the lack of a history of severe violence, or by the presence of few classic risk factors for homicide. Efforts to reduce femicide risk that are targeted only at those women seeking help for violence-related problems may miss potential victims. PMID- 14521641 TI - Desire for antibiotics and antibiotic prescribing for adults with upper respiratory tract infections. AB - OBJECTIVE: Prior studies have shown that 60% to 75% of adults with upper respiratory tract infections want antibiotics. More recent research indicates declines in antibiotic prescribing for upper respiratory tract infections. To investigate whether there has been a comparable decrease in patients' desire for antibiotics, we measured the proportion of adults with upper respiratory tract infections who wanted antibiotics in the winter of 2001-2002. We also sought to identify factors independently associated with wanting antibiotics and antibiotic prescribing. DESIGN: Prospective survey of adults with upper respiratory tract infections prior to visiting an acute care clinic from November 2001 to February 2002. MEASUREMENTS AND MAIN RESULTS: Thirty-nine percent of 310 patients wanted antibiotics. Many patients wanted relief from symptoms (43%) or pain (24%) and many patients expected to receive a diagnosis (49%) or reassurance during the visit (13%). In multivariable modeling, independent predictors of wanting antibiotics were prior antibiotic use (odds ratio [OR], 2.4; 95% confidence interval [CI], 1.3 to 4.7) and current smoking (OR, 3.1; 95% CI, 1.3 to 7.3). Physicians prescribed antibiotics to 46% of patients who wanted antibiotics and 29% of patients who did not want antibiotics (P=.01). In multivariable modeling, wanting antibiotics was an independent predictor of antibiotic prescribing (OR, 2.1; 95% CI, 1.1 to 4.4). CONCLUSIONS: Only 39% of adults seeking care for upper respiratory tract infections wanted antibiotics, less than in previous studies. In continuing efforts to break the cycle of inappropriate antibiotic use, physicians should not assume that most patients with upper respiratory tract infections want antibiotics. PMID- 14521642 TI - Health-related quality of life of adults with upper respiratory tract infections. AB - OBJECTIVE: To determine the impact that upper respiratory tract infections have on patients' physical, social, and emotional functioning, we measured the health related quality of life (HRQL) of adults with upper respiratory tract infections. SETTING: Acute care clinic from November 2001 to February 2002. DESIGN: Prospectively administered survey. To measure HRQL, we used the Acute Form of the Short Form-36, version 2 (SF-36). For all 8 SF-36 subscales, we used norm-based scoring, in which the general U.S. population has a mean of 50. PATIENTS: Adults who had symptoms for fewer than 30 days completed the SF-36; and were diagnosed with nonspecific upper respiratory infection, viral syndrome, otitis media, sinusitis, nonstreptococcal pharyngitis, streptococcal pharyngitis, or acute bronchitis. MEASUREMENTS AND MAIN RESULTS: The sample of 318 patients was 63% female, 81% white, and had a mean age of 35 years. The primary diagnoses were nonspecific upper respiratory infection (42%), acute bronchitis (16%), sinusitis (12%), viral syndrome (9%), nonstreptococcal pharyngitis (8%), otitis media (7%), and streptococcal pharyngitis (6%). Patients had a mean general health subscale score of 50.9, which is not significantly different from the mean population value of 50 (P =.09). However, there were significant decrements in the remaining 7 subscales of the SF-36: physical functioning (45.5), role-physical (38.5), bodily pain (42.6), vitality (40.8), social functioning (37.8), role-emotional (46.8), and mental health (46.8; P <.0001 for all 7 subscales compared with normative values). Results were similar for the subset of patients with no comorbid illnesses (P <.001 for the same 7 subscales) and patients diagnosed with nonspecific upper respiratory infection (P <.001 for the same 7 subscales). These decrements were similar in magnitude, but somewhat different in subscale pattern, to those of adults with chronic lung disease, osteoarthritis, and depression. CONCLUSIONS: Physicians should remember that adults who seek care for upper respiratory tract infections have measurable, significant decrements in HRQL. For researchers, HRQL is an attractive, potential measure of outcome in future trials of established and novel therapies for upper respiratory tract infections. PMID- 14521643 TI - Management of influenza symptoms in healthy adults. AB - OBJECTIVE: To determine the cost-effectiveness of rapid diagnostic testing and empiric antiviral therapy for healthy adults with symptoms of influenza. DESIGN: Cost-effectiveness analysis using a decision model based on previously published data. Outcome measures included costs and quality-adjusted life expectancy. SETTING: Physician's office. PATIENTS/PARTICIPANTS: Hypothetically healthy, working adults < 65 years of age presenting with cough and fever during the influenza season. INTERVENTIONS: Rapid testing or clinical diagnosis followed by treatment with amantadine, rimantadine, oseltamivir, or zanamivir compared with no antiviral therapy. RESULTS: Base-case analysis: not giving antiviral therapy is the most expensive and least effective strategy, costing 471 dollars per patient, mostly owing to time lost from work. Amantadine treatment increases life expectancy by 0.0014 quality-adjusted life years (QALYs) while saving 108 dollars per patient relative to no antiviral therapy. Zanamivir is slightly more effective than amantadine, adding 0.0002 QALYs at an incremental cost of 31 dollars, or 133,000 dollars per QALY saved. All other strategies, including testing strategies, are both less effective and more expensive. SENSITIVITY ANALYSIS: The model is sensitive to the probability of influenza infection, proportion of influenza caused by type B, the relative efficacy of the various drugs, and the value of a workday. At a clinical probability of influenza infection > 20%, antiviral therapy is favored. As the proportion of influenza B increases, zanamivir is favored over amantadine. Testing is rarely indicated. Ignoring the costs of lost workdays, amantadine treatment costs 1,200 dollars/QALY saved. CONCLUSIONS: Antiviral therapy with either amantadine or zanamivir is cost-effective for healthy, young patients with influenza-like illness during the influenza season, depending on the prevalence of influenza B. PMID- 14521644 TI - Physician awareness and self-reported use of local and national guidelines for community-acquired pneumonia. AB - OBJECTIVES: To assess physician awareness and reported use of medical guidelines for community-acquired pneumonia (CAP), and to identify factors associated with variations in awareness and use of these guidelines. DESIGN: A questionnaire was administered during the preintervention phase of a randomized clinical trial of a pneumonia guideline implementation strategy. PARTICIPANTS: Three hundred and fifty-two physicians who managed CAP patients at 7 Pittsburgh, PA hospitals completed the questionnaire. Physician and practice setting characteristics, and physician awareness and reported use of national American Thoracic Society (ATS) and local (hospital-developed) guidelines for CAP were assessed. RESULTS: Overall, 48% reported being influenced by ATS guidelines and 20% reported using these guidelines; 48% were uncertain whether a local pneumonia guideline existed. Only 28% of physicians who knew a local guideline existed reported frequently using the guideline. Use of national ATS guidelines was independently associated with practice as an infectious disease or pulmonary medicine specialist, nonpatient care-related professional activities, and intellect personality score. Use of local guidelines was independently negatively associated with practice as an infectious disease or pulmonary medicine specialist, and positively associated with positive attitudes toward practice guidelines. CONCLUSIONS: Results indicate low levels of awareness and use of guidelines for the management of CAP. Key indicators (e.g., medical specialty, fewer clinical duties, and positive attitudes about guidelines) were associated with greater use of national and local guidelines. If replicated with data on actual physician management practices, more effective guideline implementation strategies will be necessary to encourage compliance with practice guidelines for the management of CAP. PMID- 14521645 TI - APACHE II predicts long-term survival in COPD patients admitted to a general medical ward. AB - OBJECTIVE: The Acute Physiology and Chronic Health Evaluation II (APACHE II) was developed to predict intensive-care unit (ICU) resource utilization. This study tested APACHE II's ability to predict long-term survival of patients with chronic obstructive pulmonary disease (COPD) admitted to general medical floors. DESIGN: We performed a retrospective cohort study of patients admitted for COPD exacerbation outside the ICU. APACHE II scores were calculated by chart review. Mortality was determined by the Social Security Death Index. We tested the association between APACHE II scores and long-term mortality with Cox regression and logistic regression. PATIENTS: The analysis included 92 patients admitted for COPD exacerbation in two Burlington, Vermont hospitals between January 1995 and June 1996. MEASUREMENTS AND MAIN RESULTS: In Cox regression, APACHE II score (hazard ratio [HR] 1.76 for each increase in a 3-level categorization, 95% confidence interval [CI] 1.16 to 2.65) and comorbidity (HR 2.58; 95% CI, 1.36 to 4.88) were associated with long-term mortality (P <.05) in the univariate analysis. After controlling for smoking history, comorbidity, and admission pCO2, APACHE II score was independently associated with long-term mortality (HR 2.19; 95% CI, 1.27 to 3.80). In univariate logistic regression, APACHE II score (odds ratio [OR] 2.31; 95% confidence internal [CI] 1.24 to 4.30) and admission pCO2 (OR 4.18; 95% CI, 1.15 to 15.21) were associated with death at 3 years. After controlling for smoking history, comorbidity, and admission pCO2, APACHE II score was independently associated with death at 3 years (OR 2.62; 95% CI, 1.12 to 6.16). CONCLUSION: APACHE II score may be useful in predicting long-term mortality for COPD patients admitted outside the ICU. PMID- 14521646 TI - Interactive faculty development seminars improve the quality of written feedback in ambulatory teaching. AB - We performed a pre-post study of the impact of three 90-minute faculty development workshops on written feedback from encounters during an ambulatory internal medicine clerkship. We coded 47 encounters before and 43 after the workshops, involving 9 preceptors and 44 third-year students, using qualitative and semiquantitative methods. Postworkshop, the mean number of feedback statements increased from 2.8 to 3.6 statements (P =.06); specific (P =.04), formative (P =.03), and student skills feedback (P =.01) increased, but attitudinal (P =.13) and corrective feedback did not (P =.41). Brief, interactive, faculty development workshops may refine written feedback, resulting in more formative specific written feedback comments. PMID- 14521647 TI - "They treated me like a leper". Stigmatization and the quality of life of patients with hepatitis C. AB - OBJECTIVE: Hepatitis C virus is the most prevalent chronic blood-borne infection in the United States, typically acquired through contaminated blood products or needle sharing. We hypothesized that patients with chronic hepatitis C infection experience stigmatization independent of mode of acquisition and that it negatively affects quality of life. DESIGN: Cross-sectional observation study. SETTING: Specialty clinic in a tertiary referral hospital. PATIENTS: Two hundred and ninety outpatients diagnosed with chronic hepatitis C infection and seen in a hepatology clinic. Thirty participants were excluded because of missing data. MEASUREMENTS AND MAIN RESULTS: Patients were asked to complete a demographic profile, a semistructured interview, the Sickness Impact Profile, and the Hospital Anxiety Depression Scale. A team of two blinded coders analyzed the interviews. A total of 147 of the 257 study patients experienced stigmatization that they attributed to the disease. Women were more likely to report perceived stigmatization than men (P <.05). Age, education, professional status, and mode of infection did not influence the likelihood of stigmatization. Stigmatization was associated with higher anxiety (P <.01) and depression (P <.01), worsened quality of life (P <.01), loss of control (P <.01), and difficulty coping (P <.01). Individuals who experienced stigmatization also mentioned problems in their health care (P <.01) and work environment (P <.01) as well as with family members (P <.01). CONCLUSION: Stigmatization is a very common emotionally burdensome experience for patients with hepatitis C, which can erode social support. As it penetrates even into the health care environment, physicians and other care providers should be aware of the existence and impact of such negative stereotyping. PMID- 14521648 TI - Racial differences in initial treatment for clinically localized prostate cancer. Results from the prostate cancer outcomes study. AB - OBJECTIVE: We examined whether there were racial differences in initial treatment for clinically localized prostate cancer and investigated whether demographic, socioeconomic, clinical, or tumor characteristics could explain any racial differences. DESIGN: Prospective cohort study. SETTING: Population-based tumor registries in Connecticut, Los Angeles, and Atlanta. PARTICIPANTS: We evaluated 1144 African-American and non-Hispanic white men, aged 50 to 74 years, with clinically localized cancer diagnosed between October 1994 and October 1995. MEASUREMENTS AND MAIN RESULTS: We obtained demographic, socioeconomic, and clinical data from patient surveys and medical record abstractions. We reported adjusted percentages for receiving treatment derived from multinomial logistic regression. We found an interaction between race and tumor aggressiveness. Among men with more aggressive cancers (PSA > or = 20 ng/mL or Gleason score > or = 8), African Americans were less likely to undergo radical prostatectomy than non Hispanic whites (35.2% vs 52.0%), but more likely to receive conservative management (38.9% vs 16.3%, P=.003). Among the 71% of subjects with less aggressive cancers, African Americans and non-Hispanic whites were equally likely to receive either radical prostatectomy or radiation therapy (80.0% vs 84.5%, P=.2). CONCLUSIONS: African Americans with more aggressive cancers were less likely to undergo radical prostatectomy and more likely to be treated conservatively. These treatment differences may reflect African Americans' greater likelihood for presenting with pathologically advanced cancer for which surgery has limited effectiveness. Among men with less aggressive cancers-the majority of cases-there were no racial differences in undergoing radical prostatectomy or radiation therapy. PMID- 14521649 TI - Review of evidence and explanations for suboptimal screening and treatment of dyslipidemia in women. A conceptual model. AB - Screening and treatment rates for dyslipidemia in populations at high risk for cardiovascular disease (CVD) are inappropriately low and rates among women may be lower than among men. We conducted a review of the literature for possible explanations of these observed gender differences and categorized the evidence in terms of a conceptual model that we describe. Factors related to physicians' attitudes and knowledge, the patient's priorities and characteristics, and the health care systems in which they interact are all likely to play important roles in determining screening rates, but are not well understood. Research and interventions that simultaneously consider the influence of patient, clinician, and health system factors, and particularly research that focuses on modifiable mechanisms, will help us understand the causes of the observed gender differences and lead to improvements in cholesterol screening and management in high-risk women. For example, patient and physician preferences for lipid and other CVD risk factor management have not been well studied, particularly in relation to other gender-specific screening issues, costs of therapy, and by degree of CVD risk; better understanding of how available health plan benefits interact with these preferences could lead to structural changes in benefits that might improve screening and treatment. PMID- 14521651 TI - Interpersonal violence: improving victim recognition and treatment. PMID- 14521652 TI - Prostate cancer and race: variation in diagnosis and treatment. PMID- 14521650 TI - The long-term health outcomes of childhood abuse. An overview and a call to action. AB - While the association between abuse in childhood and adverse adult health outcomes is well established, this link is infrequently acknowledged in the general medical literature. This paper has 2 purposes: (1) to provide a broad overview of the research on the long-term effects of child abuse on mental and physical health including some of the potential pathways, and (2) to call for collaborative action among clinicians, psychosocial and biomedical researchers, social service agencies, criminal justice systems, insurance companies, and public policy makers to take a comprehensive approach to both preventing and dealing with the sequelae of childhood abuse. PMID- 14521653 TI - Prolonged atrial action potential durations and polymorphic atrial tachyarrhythmias in patients with long QT syndrome. AB - Atrial APD and Polymorphic AT in LQTS. INTRODUCTION: Prolongation of the QT interval and torsades de pointes tachycardias due to altered expression or function of repolarizing ion channels are the hallmark of congenital long QT syndrome (LQTS). The same ion channels also contribute to atrial repolarization, and familial atrial fibrillation may be associated with a mutated KVLQT1 gene. We therefore assessed atrial action potential characteristics and atrial arrhythmias in LQTS patients. METHODS AND RESULTS: Monophasic action potentials (MAPs) were simultaneously recorded from the right atrial appendage and the inferolateral right atrium in 10 patients with LQTS (8 with identifiable genotype) and compared to 7 control patients. Atrial arrhythmias also were compared to MAPs recorded in patients with persistent (n = 10) and induced (n = 4) atrial fibrillation. Atrial action potential durations (APD) and effective refractory periods (ERP) were prolonged in LQTS patients at cycle lengths of 300 to 500 msec (APD prolongation 30-41 msec; ERP prolongation 26-52 msec; all P < 0.05). Short episodes of polymorphic atrial tachyarrhythmias (polyAT, duration 4-175 sec) occurred spontaneously or during pauses after pacing in 5 of 10 LQTS patients, but not in controls (P < 0.05). P waves showed undulating axis during polyAT. Cycle lengths of polyAT were longer than during persistent and induced atrial fibrillation. Afterdepolarizations preceded polyAT in 2 patients. The electrical restitution curve was shifted to longer APD in LQTS patients and to even longer APD in LQTS patients with polyAT. CONCLUSION: This group of LQTS patients has altered atrial electrophysiology: action potentials are prolonged, and polyAT occurs. PolyAT appears to be a specific arrhythmia of LQTS reminiscent of an atrial form of "torsades de pointes". PMID- 14521654 TI - Atrial arrhythmias in the inherited long QT syndrome: laboratory quirk or clinical arrhythmia? PMID- 14521655 TI - Distal right ventricular coil position reduces defibrillation thresholds. AB - Distal RV Coil Position Reduces DFTs. INTRODUCTION: Understanding the factors that affect defibrillation thresholds (DFTs) has important implications both for optimization of defibrillation efficacy and for the design of new transvenous leads. The aim of this prospective study was to test the hypothesis that defibrillation efficacy is improved with the right ventricular (RV) coil in a distal position compared with a more proximal RV coil position. METHODS AND RESULTS: A novel defibrillation lead with three adjacent RV defibrillation coils (distal 0.8 cm, middle 3.7 cm, proximal 0.8 cm) was used for this study to permit comparison of DFTs with the proximal and distal RV coil positions without lead repositioning. In the distal RV configuration, the distal and middle RV coils were connected electrically as the anode for defibrillation. In the proximal RV configuration, the middle and proximal coils were the anode. A superior vena cava (SVC) coil and active can were connected electrically as the cathode (reversed polarity, RV-->Can+SVC). In each patient, the DFT was measured twice using a binary search protocol with the distal RV and proximal RV configurations, with the order of testing randomized. The study cohort consisted of 31 subjects (mean age 65 +/- 12 years, mean left ventricular ejection fraction 30% +/- 16%, 81% male predominance). The mean delivered energy (8.2 +/- 5.3 J vs 11.2 +/- 6.1 J), leading-edge voltage (335 +/- 109 V vs 393 +/- 118 V), and peak current (11.6 +/- 5.2 A vs 14.9 +/- 7.3 A) at DFT all were significantly lower with the distal RV configuration compared to the proximal RV configuration (P < 0.01 for all comparisons). CONCLUSION: DFTs are significantly reduced with the distal RV configuration compared to the proximal RV configuration. Defibrillation leads should be designed with the shortest tip to coil distance that can be achieved without compromising ventricular fibrillation sensing. PMID- 14521656 TI - Global right atrial mapping delineates double posterior lines of block in patients with typical atrial flutter: a study using a three dimensional noncontact mapping system. AB - Double Posterior Lines of Block in Typical Atrial Flutter. INTRODUCTION: The crista terminalis (CT) has been shown to be a barrier to transverse conduction during typical atrial flutter (AFL). However, some studies have demonstrated the presence of functional block in the sinus venosa region but not at the CT. The aim of this study was to define these regions of block in the right atrium using a three-dimensional noncontact mapping system. METHODS AND RESULTS: In 39 AFL patients (33 men and six women, mean age 56 +/- 13 years), a noncontact multielectrode array was used to reconstruct electrograms in the right atrium. Isochronal and isopotential propagation mapping was performed during AFL and during pacing from the coronary sinus ostium and the low lateral wall (cycle length from 600 to 240 msec) in sinus rhythm after creation of isthmus block. A single line of block along the CT area was found in 18 patients (46%). Two lines of block were found in 21 patients (54%), with the first line located along the CT area. The second was located in the sinus venosa region in 20 patients (51%) and in the lateral wall in 1 patient (3%). In all patients, the block in the lower part of the CT was observed during AFL (60%) and during pacing at all cycle lengths (48%-62%). The length and proportion of block were inversely proportional to pacing cycle length. CONCLUSION: Double lines of block were frequently observed in patients with AFL, and both lines may form the posterior boundaries of the AFL circuit. Block was fixed in the lower part of the CT and was functional in the upper part of the CT. PMID- 14521657 TI - Clinical significance and contributing factors of long-term variability in induced ventricular tachyarrhythmias. AB - Long-Term Variability in Induced Tachyarrhythmias. INTRODUCTION: Ventricular arrhythmias induced during electrophysiologic study (EPS) may vary over time, making arrhythmia induction studies unreliable. The aim of this prospective study was to clarify the clinical significance of long-term variability in induced arrhythmias and to elucidate factors determining this variability. METHODS AND RESULTS: Three noninvasive EPSs were performed 1, 13, and 25 months after implantation of a cardioverter defibrillator in 40 patients with ventricular tachyarrhythmias, without a change in their antiarrhythmic drug regimens. The induced ventricular arrhythmias were categorized into five grades. Long-term variability, which was defined as a variation in the grades during the three EPSs, was observed in 23 patients (group A) and not in the remaining 17 patients (group B). During the 2-year period, spontaneous sustained ventricular arrhythmias developed in 15 patients (65%) in group A but in only 4 patients (24%) in group B (P = 0.01). Inducibility of sustained tachyarrhythmias was not associated with emergence of spontaneous arrhythmias. All patients also underwent thallium-201 and iodine-123-metaiodobenzylguanidine (MIBG) scans to evaluate the extent of the regions showing normal thallium uptake with reduced MIBG uptake. Group A patients showed greater thallium/MIBG mismatched regions than did group B patients (P = 0.01). Logistic regression analysis revealed that long-term variability (relative risk [RR] 7.55, P = 0.03), amiodarone therapy (RR 0.14, P = 0.04), and left ventricular ejection fraction <35% (RR 6.26, P = 0.04) were independent predictors of spontaneous arrhythmia occurrence. CONCLUSION: In patients with ventricular tachyarrhythmias, long-term variability in induced arrhythmias, but not the inducibility of arrhythmias, is associated with a higher incidence of spontaneous arrhythmias. Regional cardiac sympathetic denervation may be an important mechanism of this variability. These results also may explain why inducibility-based antiarrhythmic drug testing does not predict patient prognosis. PMID- 14521658 TI - Fasciculoventricular pathways: clinical and electrophysiologic characteristics of a variant of preexcitation. AB - Fasciculoventricular Fibers. INTRODUCTION: Fasciculoventricular tracts are considered a rare form of ventricular preexcitation. Few fasciculoventricular pathways have been reported, and none have been linked to a reentrant tachycardia. METHODS AND RESULTS: Four patients with fasciculoventricular bypass tracts underwent electrophysiologic evaluation. Two patients had a single fasciculoventricular pathway, one that inserted anteroseptally and the other in the left ventricle. Two patients also had an AV bypass tract, with anterograde conduction over the fasciculoventricular pathway during orthodromic AV reentrant tachycardia. After ablation of the AV pathways, the ECG during sinus rhythm and the electrophysiologic study showed ventricular preexcitation due to a fasciculoventricular bypass tract inserting into the right ventricle. Adenosine triphosphate was helpful in the diagnostic process. CONCLUSION: Electrophysiologists should be able to make the differential diagnosis between a fasciculoventricular bypass tract and an anteroseptal accessory pathway to preclude potential harm to the AV conduction system if a fasciculoventricular pathway is targeted for catheter ablation. PMID- 14521659 TI - Interactions between extracellular stimuli and excitation waves in an atrial reentrant loop. AB - Extracellular Stimuli in an Atrial Reentrant Loop. INTRODUCTION: The interactions between extracellular stimuli and excitation waves propagating in a reentrant loop are a complex function of stimulus parameters, structural properties, membrane state, and timing. Here the goal was a comprehensive understanding of the mechanisms and frequencies of the major interactions between the advancing excitation wave and a single extracellular stimulus, separated from issues of anatomic or geometric complexity. METHODS AND RESULTS: A modernized computer model of a thin ring of uniform tissue that included a pair of extracellular stimulus electrodes (anode/cathode) was used to model one-dimensional cardiac reentry. Questions and results included the following: (1) What are the major interactions between a stimulus and the reentrant propagation wave, and are they induced near the cathode or near the anode; and, for each interaction, what are the initiating amplitude range and timing interval? At the cathode, the well known mechanism of retrograde excitation terminated reentry; changes in timing or amplitude produced double-wave reentry or phase reset. At the anode, termination occurred at different cells depending on stimulus amplitude. (2) Relatively how often did termination occur at the anode? For most stimulus amplitudes, termination occurred more often at the anode than at the cathode, although not always at the same cell. (3) With random timing, what is the probability of terminating reentry? Stimulation for 5 msec terminated reentry with a probability from 0% to approximately 10%, as a function of increasing stimulus amplitude. CONCLUSION: A single extracellular stimulus can initiate major changes in reentrant excitation via multiple mechanisms, even in a simple geometry. Termination of reentry, phase shifts, or double-wave reentry each occurs over well-defined ranges of stimulus amplitude and timing. PMID- 14521660 TI - Responses to pacing depend on electrical and structural properties of the reentrant circuit. PMID- 14521662 TI - Mapping a moving target. PMID- 14521661 TI - Effects of cytochalasin D on electrical restitution and the dynamics of ventricular fibrillation in isolated rabbit heart. AB - Cytochalasin D in Rabbit Ventricle. INTRODUCTION: Cytochalasin D (cyto-D) has been used as an excitation-contraction uncoupler during optical mapping studies. However, its effects on action potential duration restitution (APDR) and dynamics during ventricular fibrillation (VF) are unclear. METHODS AND RESULTS: Langendorff-perfused rabbit hearts (N = 6) were immersed in a tissue chamber. Transmembrane potential was recorded using glass microelectrodes. APD measured to 90% repolarization (APD90) was used to construct the APDR curve. During regular pacing at 300-msec cycle length, increasing concentrations of cyto-D resulted in progressively prolonged APD90 (131 +/- 26 msec, 171 +/- 14 msec, and 177 +/- 14 msec) and steepened maximum slope of the APDR curve (1.1 +/- 0.2, 1.3 +/- 0.2, and 1.6 +/- 0.4 for control, 5 micromoles, and 10 micromoles, respectively; P < 0.01). Resting membrane potential, AP amplitude, and maximum dV/dt did not change. Cyto-D lengthened VF cycle length and APD90, and steepened the maximum slope of the APDR curve. However, cyto-D did not significantly change the diastolic interval. The dominant frequency of pseudoelectrocardiogram progressively decreased with increasing concentrations of cyto-D (15.2 +/- 0.6 Hz, 11.1 +/- 2.4 Hz, and 9.8 +/- 3.2 Hz for control, 5 micromoles, and 10 micromoles, respectively; P < 0.01). Sustained (>1 min) VF was repeatedly inducible at baseline and with 5 or 10 micromoles of cyto-D. CONCLUSION: Continuous perfusion of cyto-D at 5 or 10 micromoles prolonged APD90, steepened APDR slope, and reduced dominant frequency in rabbit ventricles. Cyto-D at these concentrations allowed induction of sustained VF. PMID- 14521663 TI - Epicardium-based left atrial ablation: impact on electromechanical properties. AB - Epicardium-Based LA Ablation. INTRODUCTION: An important developmental task for surgical ablation of atrial fibrillation is simplification. A significant step would be the development of tools that create satisfactory ablation lesions without necessitating cardiopulmonary bypass or atriotomy. Optimally, these lesions would have no adverse impact on atrial electromechanical properties. METHODS AND RESULTS: We sought to characterize left atrial (LA) lesions created by a bipolar ablation device without cardiopulmonary bypass or atriotomy and to assess their impact on LA electromechanical properties. In each of five pigs, lesions were delivered to the epicardium of the beating heart, and encircled and connected right and left pulmonary vein vestibules and the atrial appendage. Before and after ablation, LA electromechanical properties were assessed using endocardial electromechanical mapping and intracardiac echocardiography. Postmortem histologic analysis also was performed. Each lesion was thrombus-free and barotrauma-free, histologically transmural, and a complete conduction barrier. Although a large aggregate area (24% +/- 6%) of LA myocardium was excluded, there was no significant change in global electromechanical properties. However, marked diminishment in appendage function was observed. CONCLUSION: Epicardium-based LA ablation in a beating heart could be achieved successfully without cardiopulmonary bypass or atriotomy. Although there was no adverse impact on global electromechanical properties, there was evidence of important regional diminishment. PMID- 14521664 TI - Focal origin of atrial tachycardia in dogs with rapid ventricular pacing-induced heart failure. AB - Mapping and Ablation of Atrial Tachycardia in Heart Failure. INTRODUCTION: Dogs with rapid ventricular pacing-induced congestive heart failure (CHF) have inducible atrial tachycardia (AT), with a mechanism consistent with delayed afterdepolarization-mediated triggered activity. We assessed the hypothesis that AT has a focal origin. METHODS AND RESULTS: Twenty-one CHF dogs undergoing 3 to 4 weeks of ventricular pacing at 235 beats/min were studied. Biatrial epicardial mapping of 20 sustained AT episodes (cycle length [CL], 175 +/- 53 msec) in 5 dogs revealed an area of earliest activation in the right atrial (RA) free wall (13 episodes), RA appendage (4 episodes), or between the pulmonary veins (3 episodes). Total epicardial activation time during AT (73 +/- 19 msec) was similar to that during sinus rhythm (72 +/- 13 msec) and on average was <50% of the AT CL. Higher-density mapping of the RA free wall during 30 sustained AT episodes (163 +/- 55 msec) in 9 dogs identified a site of earliest activation along the sulcus terminalis most frequently as a stable, focal activation pattern from a single site. Endocardial mapping of 49 sustained AT episodes (156 +/- 27 msec) in 10 dogs revealed multiple sites of AT origin arising along the crista terminalis and pulmonary veins. Right and left ATs were terminated with discrete radiofrequency ablation, but other ATs remained inducible. A rapid, left AT generating an ECG pattern of atrial fibrillation was ablated inside the pulmonary vein. CONCLUSION: AT induced in this CHF model after 3 to 4 weeks of rapid ventricular pacing has an activation pattern consistent with a focal origin. Sites of earliest activation are distributed predominately along the crista terminalis and within or near the pulmonary veins. PMID- 14521665 TI - Spatiotemporal correlation between phase singularities and wavebreaks during ventricular fibrillation. AB - Phase Singularity and Wavebreak. INTRODUCTION: Phase maps and the detection of phase singularities (PSs) have become a well-developed method for characterizing the organization of ventricular fibrillation (VF). How precisely PS colocalizes with wavebreak (WB) during VF, however, is unknown. METHODS AND RESULTS: We performed optical mapping of 27 episodes of VF in nine Langendorff-perfused rabbit hearts. A WB is a point where the activation wavefront and the repolarization waveback meet. A PS is a site where its phase is ambiguous and its neighboring pixels exhibit a continuous phase progression from -pi to +pi. The correlation coefficient between the number of WBs and PSs was 0.78 +/- 0.09 for each heart and 0.81 for all VF episodes (P < 0.001), indicating a significant temporal correlation. We then superimposed the WBs and PSs for every 100 frames of each episode. These maps showed a high degree of spatial colocalization. To quantify spatial colocalization, the spatial shifts between the cumulative maps of WBs and PSs in corresponding frames were calculated by automatic alignment to obtain maximum overlap between these two maps. The spatial shifts were 0.04 +/- 0.31 mm on the x-axis and 0.06 +/- 0.27 mm on the y-axis over a 20 x 20 mm2 mapped field, indicating highly significant spatial correlation. CONCLUSION: Phase mapping provides a convenient and robust approach to quantitatively describe wave propagation and organization during VF. The close spatiotemporal correlation between PSs and WBs establishes that PSs are a valid alternate representation of WB during VF and further validated the use of phase mapping in the study of VF dynamics. PMID- 14521666 TI - Transvenous biventricular pacing in a child after congenital heart surgery as an alternative therapy for congestive heart failure. AB - Transvenous Biventricular Pacing in Children. Cardiac resynchronization therapy improves short-term and long-term hemodynamics in adult patients with congestive heart failure and left bundle branch block. We describe the feasibility of transvenous biventricular pacemaker implantation in a 6-year-old child with heart failure and wide QRS complex after congenital heart surgery. Myocardial tissue Doppler imaging was used to demonstrate intraventricular dyssynchrony and resynchronization after cardiac resynchronization therapy. During 1-year follow up, symptomatology and hemodynamic parameters improved. PMID- 14521667 TI - Risk stratification and management of sudden cardiac death: a new paradigm. AB - Risk Stratification and Management of SCD. Management of SCD is undergoing radical change in direction. It is becoming increasingly appreciated that besides depressed left ventricular systolic function and the conventional risk stratification tools, new markers for plaque vulnerability, enhanced thrombogenesis, specific genetic alterations of the autonomic nervous system, cardiac sarcolemmal and contractile proteins, and familial clustering may better segregate patients with atherosclerotic coronary artery disease who are at high risk for SCD from those who may suffer from nonfatal ischemic events. Better understanding of pathophysiologic processes, such as postmyocardial infarction remodeling, the transition from compensated hypertrophy to heart failure, and the increased cardiovascular risk of coronary artery disease in the presence of diabetes or even a prediabetic state will help to improve both risk stratification and management. The rapidly developing fields of microchips technology and proteomics may allow rapid and cost-effective mass screening of multiple risk factors for SCD. The ultimate goal is to identify novel methods for risk stratification, risk modification, and prevention of SCD that could be applied to the general public at large. PMID- 14521669 TI - Should we control rate or rhythm in atrial fibrillation? PMID- 14521668 TI - How really rare are rare diseases?: the intriguing case of independent compound mutations in the long QT syndrome. PMID- 14521670 TI - Termination of a narrow QRS-complex tachycardia: what is the mechanism? PMID- 14521671 TI - Supraventricular tachycardia with sudden rate doubling: what is the mechanism? PMID- 14521672 TI - Electroanatomic mapping of human heart: epicardial fat can mimic scar. PMID- 14521674 TI - Implantable cardioverter defibrillator in high-risk long QT syndrome patients. PMID- 14521676 TI - To the editor. PMID- 14521677 TI - Electrophysiology of ventricular tachycardia: an historical perspective. PMID- 14521679 TI - Praiseworthy pragmatism? Validity and action research. AB - BACKGROUND: Action research has been promoted as a suitable methodology for nursing, but is located in a discourse that challenges its claims of validity. This discourse, influenced by positivism, is similar to that which qualitative researchers have faced. AIM: This article contributes to theoretical discussion of the validity of action research. FINDINGS: Because of similarities, the emergent debates around the reconceptualization of validity in qualitative research are outlined, prior to a consideration of the relative arguments in relation to action research. Three avenues of thought are discerned: that no criteria are required; that parallels with other schools are appropriate; and that new concepts specific to one school of thought should be developed. CONCLUSIONS: Because of its ethical and action-oriented principles, action research offers the potential to address validity issues that remain problematic to 'pure' qualitative researchers, and it can be claimed that its pragmatic approach is validity-enhancing. We argue for the rejection of naive rule-based formulae and for recognition of the impact of contextual and pragmatic concerns, so that the potential for the added value of action research might be realized. PMID- 14521680 TI - Pressure ulcer risk assessment: application of logistic analysis. AB - AIM: The aim of this study was to investigate the relative importance of key factors affecting the likelihood of hospital acquired pressure ulcers, thus offering the groundwork for the development of an empirically-based risk assessment scale. It also evaluated the predictive performance of the underlying empirical model and compared its classification ability with the empirically observed ability of the Waterlow risk assessment scale. BACKGROUND: Pressure ulcers impose a significant burden on patients and carers and demand substantial resources from the health care system. There is, however, a lack of systematic empirical research on which to base the development of risk assessment measurement tools. METHODS: Multivariate statistical methods were applied to data derived from the records of a cross-sectional sample of around 500 randomly selected inpatient episodes drawn from the population of admissions to a single unit in a large acute hospital during a 2-year period (October 1996 to September 1998). Double-checking of a randomly selected 25% sample of the original records and careful screening out of records with incomplete information or inconsistent values was carried out to ensure a high quality sample. Logit analysis was used to investigate the relative contribution of risk factors, such as continence, skin condition, mobility and inter-hospital transfer to the risk of hospital acquired pressure ulcer occurrence, whilst cross-validation techniques were employed to check the predictive performance of the model. RESULTS: The results suggest that a simplified version of the Waterlow risk assessment tool has satisfactory predictive ability and the potential for further development. CONCLUSIONS: Two main conclusions emerged from this study. First, it is both feasible and worthwhile to pursue improvement in the development of risk assessment tools using statistical methods. Second, locally-determined risk factors will need to be incorporated into the construction of future risk assessment scales. PMID- 14521681 TI - Nursing interventions for patients with chronic conditions. AB - BACKGROUND: Maintaining independence in older people or restoring and encouraging self-management in those with chronic conditions are ways of increasing health related quality of life and at the same time may achieve cost savings. Nurses often carry out these types of interventions in follow-up home visits. AIMS: To describe nursing interventions during home visits and their effects on people suffering from a range of chronic conditions. METHOD: A structured descriptive review was carried out using the Medline, Embase, PsycINFO, CINAHL and Cochrane databases and the terms chronic or diabetic or arthrit* or pain and randomi* and [(nurs* and care) or (nurs* and interventions)]. FINDINGS: In older people the best outcomes are reached if the target population is 'the younger-old', or if intervention is tailored to elders who have stated health problems. The effect depends on the duration of the follow-up period, number of follow-up visits and personality of the nurse. Follow-up nursing interventions for patients with diabetes can improve psychosocial and health outcomes. Education of health providers and amount of time spent with patients seem to be important for positive outcome. A rheumatology nurse specialist seems to provide equally good clinical results as a rheumatologist and produces better patient-related outcomes. Compared with a multidisciplinary team treating patients in day care or inpatient settings, a rheumatology nurse specialist provides less patient satisfaction, but equally good clinical outcomes. CONCLUSIONS: Patients with chronic conditions often benefited from follow-up visits by nurses. Visits should be multiple, extended over a long-term, and interventions should be individualized. The outcomes may be described as patient satisfaction, good clinical outcomes and cost savings. Future randomized controlled trials should describe nurses' interventions and educational background to enable replication. Randomized controlled trials using the Health Technology Assessment method would permit comparison between studies and be a good foundation for future decision making. PMID- 14521682 TI - Effectiveness of nursing interventions in people with personality disorders. AB - BACKGROUND: Nurses usually provide care for people with personality disorders on a day-to-day basis. Consequently, it is important to establish how effective nursing interventions are for those with personality disorders, both in terms of general management and more specific therapeutic approaches. These are also issues of current political and professional debate. The crucial question, however, for planners, providers and commissioners of services is, What research evidence is there about effectiveness? There is also a need to determine what further research is necessary to evaluate programmes for managing or modifying behaviours. AIM: The aim of this paper is to report on a systematic review of the literature on the effectiveness of nursing interventions in people with personality disorders, whether delivered by nurses alone or in combination with other health care professionals. METHOD: The review was conducted according to the United Kingdom National Health Service Centre for Reviews and Dissemination guidelines. Literature was identified from electronic database searching, footnote chasing, hand searching of journals and contact with authors of studies. Narrative synthesis was undertaken in relation to study design, participants, and type of intervention. FINDINGS: Eighteen separate studies were included in the review: four randomized controlled trials, four non-randomized controlled trials, seven before-and-after studies and three case studies. CONCLUSIONS: There is a weak evidence-base for what constitutes effective nursing intervention with people with personality disorders. There is stronger evidence from mixed disciplines intervention studies than from nursing only intervention studies. Studies of interventions based on psychological approaches show greater improvements in outcomes than nursing management studies. PMID- 14521683 TI - Wandering behaviour in people with dementia. AB - BACKGROUND: Wandering has been described as one of the most challenging behaviours to manage and yet a search of the literature in the last two decades yields only a limited number of papers that specifically studied wandering. AIM: This paper reviews the current literature on wandering in relation to the nature of the phenomenon, attempts to classify the behaviour, the extent of the problem, the profile of those who wander, postulations about its aetiology, and intervention strategies being employed. METHOD: Eight literature databases were searched for the last 40 years up to February 2003 using 'wandering' and 'dementia' as combined keywords. A total of 133 articles was included in the review. FINDINGS: The prevalence of wandering behaviour was difficult to assess and no conclusions could be reached. The typical wanderer depicted in the literature was relatively young in the older population, more cognitively impaired, more likely to be a man, might have experienced sleep problems, had a more active premorbid lifestyle, and used more psychotropic medications. While studies agreed that wanderers are more cognitively impaired, their findings did not necessarily agree on other attributes. Three major approaches, namely the biomedical, psychosocial and person-environment interaction perspectives, in conceptualizing wandering behaviour can be identified from the literature. Medications, activity programmes, behavioural modification and environmental manipulation have been used as interventions but none has so far demonstrated unequivocal effectiveness. CONCLUSION: Knowledge generated through research remains insufficient to explain fully why and when wandering occurs. Variability in how the phenomenon was defined and studied, and the small size of the samples made generalizability of findings difficult. Future research should incorporate a clearer definition of wandering; a specific targeted population with representative sample size; appropriate subject identification strategies; focussed interventions, and better control conditions. PMID- 14521684 TI - Early recognition of dementia by nurses. AB - BACKGROUND: Early recognition of dementia is a key policy objective designed to maximize the efficacy of treatment and to provide timely and appropriate support before crisis occurs. The impact of early recognition of dementia is under researched in primary care nursing. AIM: To explore whether community mental health nurses (CMHNs), community nurses (CNs) and practice nurses (PNs) have different perspectives on early diagnosis of dementia and to consider the possible effects of any variation. METHOD: Data are drawn from questionnaires completed by CMHNs (79), CNs (153) and PNs (36) who attended workshops offered on 24 occasions in 21 settings across the United Kingdom. The workshops attracted a range of primary care practitioners and were part of a training programme on early diagnosis of dementia. RESULTS: Analysis of the data shows some differences in knowledge, experience and confidence between the three nurse groups. CMHNs were more confident in their abilities to recognize dementia and found providing support less difficult than CNs and PNs. CMHNs considered that they were best placed to co-ordinate services for people with newly recognized dementia. CNs and PNs, however, reported experience of working with people with dementia and many appeared able to respond to early signs and to identify potential sources of support. CONCLUSIONS: While CMHNs may have a key role in responding effectively to the newly identified needs of people with early dementia, other nurses working in the community are likely to encounter people with early dementia. In the context of a policy objective to identify people with dementia earlier, all community-based nurses should be able to recognize the possibility of dementia and support those undergoing referral or assessment. Their confidence in doing so should be enhanced by continued professional development. Training in dementia recognition, involvement in and membership of primary care teams supporting people with dementia should not be confined to CMHNs. Nurses who regularly encounter the general population of older people may be well placed to provide continuity of support for those who may, or may not, have cause to suspect that they or their relatives have early dementia. PMID- 14521685 TI - Continuing care after cancer treatment. AB - BACKGROUND: Despite nearly three decades of debate and policy guidance there is evidence that, in the United Kingdom, patient hospital discharge remains problematic. District nurses, who deliver skilled home nursing care, receive referrals from hospitals for continuing nursing care needs. However, district nurses' expectations of appropriate patient referral from hospitals are not always achieved. In an attempt to improve services after hospital discharge, government policy has emphasized partnership between care providers, highlighting the need for smooth transition between care settings. AIM: To explore hospital discharge and referral procedures for patients with cancer, with particular emphasis on referrals made by hospital nurses to district nurses. METHOD: In depth interviews were carried out with nurses actively involved in the discharge process as both referrers and recipients of referrals. Twenty nurses from a regional cancer centre and 20 district nurses from three adjacent primary care trusts were interviewed. Interviews were transcribed and analysed thematically, and themes compared between the two care settings. CONCLUSIONS: We conclude that competing sets of expectations, not only between hospital and community nursing settings, but amongst district nurses themselves, are a major factor impeding agreement on referral criteria and satisfaction with the referral process. PMID- 14521686 TI - Health visitors' understandings of domestic violence. AB - BACKGROUND: Feminist work has made visible the extent and nature of domestic violence and the problems women face in having their experiences recognized by health and welfare professionals. Research has demonstrated that many health care professionals, including nurses, midwives and health visitors have little working knowledge about this issue. This impacts on their ability to recognize and respond to domestic violence within their practice. AIM: This paper is based upon a study of British health visitors, which explored their practice in relation to domestic violence. Drawing upon empirical data from interviews with health visitors, it explores their understandings of the extent and nature of domestic violence in the context of their work. METHODS: Semi-structured interviews were undertaken with 24 health visitors selected by convenience and purposive sampling. Data collection took place during 1997-1998. The research draws on the theoretical perspectives of feminist poststructuralism. FINDINGS: The findings demonstrate considerable differences between health visitors in their understandings of the extent of domestic violence in their caseloads and their recognition of different types of abuse experienced by women. There were also differences between participants in their willingness to name situations other than physical violence as abusive, as well as the extent to which they recognized domestic violence within different social groups. CONCLUSIONS: A feminist perspective provides critical insight into the professional knowledge base in relation to domestic violence, demonstrating the need for health visitors to develop their understandings further in order to respond appropriately to women and children experiencing domestic violence. This is discussed in the context of ongoing struggles for professional identity within an ever-changing arena of health and welfare provision. PMID- 14521687 TI - Nursing and health care in Indonesia. AB - AIM: Indonesia, with its population of over 220 million, has health problems similar to those of other developing countries. In an attempt to provide nurses throughout the world with knowledge about Indonesia, we describe the country, its health system, and problems encountered by nurses and other health professionals. METHOD: We explain the way the health system works within Indonesian culture, discuss the effects of the international nursing shortage and outline the role of aid agencies. The ethical dilemmas faced by health professionals who care for patients within a poorly resourced system are examined. While the information pertains to the whole country, we focus on the main island of Java, as that is where we have worked and resided. FINDINGS: Nursing education is primarily conducted at senior high school, while medical education is similar to the university education offered in many countries, and allied health professionals are educated to varying standards. Indonesian health officials recognize that the low standard of nursing education contributes to poor health statistics, and they are working hard to improve this. There has been strong support from the government for the implementation of university education for nurses, and for courses within academies that bridge the gap between current standards and the levels of education required for the delivery of optimum health care. DISCUSSION: We both are nurses. One of us is an Indonesian and the other has worked for many years in Indonesia and coordinated a programme that organized exchanges of health professionals working in large tertiary referral hospitals and health departments in Indonesia and Australia. The information presented here is the result of many collaborative projects and gives information not available in published works. PMID- 14521688 TI - Nursing staff mix models and outcomes. AB - BACKGROUND: Recently, restructuring of the nursing workforce has been undertaken in a number of countries in an effort to provide efficient and cost-effective services to users. This often takes the form of the introduction of unregulated workers to carry out support roles with registered nurses. However, these changes have not been evaluated for efficacy or impact on nurses, patients or the health care system. PURPOSE: The purpose of this study was to determine the relationship between staff mix models comprising regulated staff (Registered Nurses and Registered Practical Nurses) or regulated and unregulated staff (Registered Nurses and unregulated workers), and nursing and quality outcomes. METHODS: This comparative correlational study was conducted in a random sample of 30 adult, acute care patient units within eight hospitals located in Toronto, Canada. Registered Nurses employed on 30 randomly selected hospital units, grouped by the two staff mix models (15 units per group), were surveyed using previously validated instruments to measure role conflict, role ambiguity, job satisfaction, perceived effectiveness of care and perceived quality of care. RESULTS: Results indicated that Registered Nurses in this study experienced high levels of role conflict, regardless of the type of staff mix model within which they worked. Registered Nurses on units employing both Registered Nurses and unregulated workers reported higher levels of job satisfaction. On units employing both Registered Nurses and unregulated workers, Registered Nurses perceived that the quality of care was lower. CONCLUSIONS: Staff mix model was related to Registered Nurses' perceptions of the quality of patient care. It was also evident that other variables within the work environment might have more influence on the outcomes examined than the independent variable of staff mix. PMID- 14521689 TI - Sweet's syndrome revisited: a review of disease concepts. AB - Sweet's syndrome, also referred to as acute febrile neutrophilic dermatosis, is characterized by a constellation of symptoms and findings: fever, neutrophilia, erythematous and tender skin lesions that typically show an upper dermal infiltrate of mature neutrophils, and prompt improvement of both symptoms and lesions after the initiation of treatment with systemic corticosteroids. Hundreds of patients with this dermatosis have been reported. The manifestations of Sweet's syndrome in these individuals have not only confirmed those originally described by Dr Robert Douglas Sweet in 1964, but have also introduced new features that have expanded the clinical and pathologic concepts of this condition. The history, clinical characteristics, laboratory findings, associated diseases, pathology, and treatment options of Sweet's syndrome are reviewed. The evolving and new concepts of this dermatosis that are discussed include: (i) Sweet's syndrome occurring in the clinical setting of a disease-related malignancy, or medication, or both; (ii) detection of additional sites of extracutaneous Sweet's syndrome manifestations; (iii) discovery of additional Sweet's syndrome-associated diseases; (iv) variability of the composition and/or location of the cutaneous inflammatory infiltrate in Sweet's syndrome lesions; and (v) additional efficacious treatments for Sweet's syndrome. PMID- 14521690 TI - Clinical and mycological status of the Trichophyton mentagrophytes (interdigitale) syndrome of chronic dermatophytosis of the skin and nails. AB - The syndrome of tinea pedis caused by human-adapted Trichophyton mentagrophytes (interdigitale) can be recognized by two signs not characteristically seen in tinea pedis caused by T. rubrum and Epidermophyton floccosum:1 bullous > 2 mm vesicles in the thin skin of the plantar arch and along the sides of the feet and heel adjacent to the thick plantar stratum corneum,2 and white superficial onychomycosis. The relationship of the syndrome to zoophilic T. mentagrophytes remains in question. PMID- 14521691 TI - Examination of Bcl-2, Bcl-X and bax protein expression in psoriasis. AB - BACKGROUND: Psoriasis is an inflammatory skin disease characterized by epidermal hyperplasia and greatly accelerated epidermal turnover. The blockage of normal apoptotic process in the epidermis is one of the factors implicated in the pathogenesis of psoriasis. OBJECTIVE: The purpose of the present study was to elucidate whether bcl-family proteins are significantly involved in the hypothetical antiapoptotic cascade in lesional psoriatic epidermis. METHODS: Twenty-six lesional biopsy samples of 26 patients with psoriasis and five control specimens from normal skin were studied by immunohistochemical method for the differential expression of pro-apoptotic bax and antiapoptotic bcl-2 and bcl-x proteins. RESULTS: Compared with the normal epidermis, bcl-2 expression was significantly reduced, whereas bax and bcl-x were significantly overexpressed in the psoriatic epidermis. The localization of bcl-2/bax/bcl-x proteins in the psoriatic epidermis did not show a significant deviation from that in the normal epidermis. CONCLUSION: These findings indicate a discordant expression of bcl-2 and bax/bcl-x in psoriatic epidermis. Increased bcl-x expression might contribute to the antiapoptotic response in psoriatic keratinocytes. The presence of strong bax expression with a concomitant decrease in bcl-2 expression suggests either a functional defect in bax protein or an inherent/acquired resistance to bax mediated apoptosis in psoriatic keratinocytes. PMID- 14521692 TI - Epidemic (human immunodeficiency virus-related) Kaposi's sarcoma in West African women. AB - BACKGROUND: Most patients infected with human immunodeficiency virus (HIV) experience skin disease at some stage of their illness, either as a presenting feature or as a later manifestation. Different dermatoses may coexist during the course of the infection, and the unusual nature of the skin lesions can make an accurate diagnosis difficult. Kaposi's sarcoma is one of the acquired immunodeficiency syndrome (AIDS)-defining skin diseases and may coexist with other lesions in AIDS patients. Kaposi's sarcoma is caused by human herpesvirus 8 (HHV8), which is mainly transmitted through male homosexual behavior, and is less common in women than in men. METHODS: The clinical, histopathologic, and therapeutic aspects of AIDS-related Kaposi's sarcoma in three women (age, 18-34 years) who presented to the dermatology clinic of the University of Nigeria Teaching Hospital, Enugu, Nigeria, were studied over 18 months, beginning in August and September 1998. RESULTS: At presentation, the condition of the three patients was generally poor, with severe anemia and extensive dark-brown papules, plaques, and nodules. Mucosal involvement was extensive. Generalized lymphadenopathy was found in two patients, one of whom had extensive coexisting dermatophytosis. All three women were in stage III/IV of the disease at the time of initial presentation. Death occurred within days in one and within 7-8 months in two. CONCLUSIONS: Because these female patients with epidemic Kaposi's sarcoma lived in an almost entirely heterosexual population, the mode of contamination with HHV8 was obscure. The course of the disease was pervasive, aggressive, and devastating. PMID- 14521693 TI - Vitiligo in the Nigerian African: a study of 351 patients in Benin City, Nigeria. AB - BACKGROUND: There is a paucity of reports on vitiligo in sub-Saharan Africa. AIM: To define the clinical and epidemiologic pattern of vitiligo in the Nigerian African. METHODS: We examined all new cases of vitiligo seen in the Dermatology Unit of the University of Benin Teaching Hospital (UBTH) over a 14-year period: January 1985 to December 1998. RESULTS: Three hundred and fifty-one patients with vitiligo, representing 3.2% of new dermatologic cases, were enrolled. The study group was made up of 153 males and 198 females, giving a sex ratio of 1 : 1.3. The peak incidence of vitiligo was in the second and third decades of life, and the most common form of presentation was the localized focal type (77%), followed by the segmental type (12.5%) and the generalized form (10.5%). Common sites affected were the limbs (32%), trunk (23.8%), face (18.2%), head and neck (9.1%), and the mucous membranes (7.4%). There was a positive family history of vitiligo in 18% of subjects and 3.4% had systemic diseases, such as diabetes mellitus (1.7%), anemia (0.8%), hyperthyroidism (0.6%), and leukemia (0.3%). CONCLUSIONS: Vitiligo on darkly pigmented skin is often a very striking disease. Our findings show that the pattern of vitiligo is similar to that reported from other parts of the world. The high level of social stigmatization is due to confusion with leprosy. PMID- 14521694 TI - Prevalence of Behcet's disease in Istanbul, Turkey. AB - BACKGROUND: The prevalence of Behcet's disease (BD) is much higher in countries along the ancient Silk Route, extending from Japan to Mediterranean countries including Turkey, than in northern Europe and the USA. Three previous epidemiologic surveys have been carried out in different regions of Turkey. PATIENTS AND METHODS: This study investigated the cross-sectional prevalence of BD in individuals aged > 12 years in Istanbul, Turkey, in two stages. The first stage aimed to identify individuals with recurrent oral ulcers (ROUs) by visiting them in their homes, and the second stage aimed to further examine those with ROUs for the presence of other BD-related manifestations under hospital conditions. The sample size was determined to be 24,000 with an expected BD prevalence rate of 1/1000 and a sampling error of 4/10,000, with a 95% confidence interval (CI) of 6-14/10,000. The number of individuals to be screened in each district was determined in proportion to the population of all districts in Istanbul. RESULTS: The standard questionnaire was applied to a total of 23,986 individuals at their homes. A history of ROU was recorded in 2289 individuals (9.5%), and a previous diagnosis of BD was recorded in 47. The diagnosis of ROU was confirmed in 700, and the diagnosis of BD was established in 101 according to the International Study Group criteria. The prevalence rate of BD was estimated as 42/10,000 (95% CI, 34-51/10,000) in Istanbul, Turkey. CONCLUSIONS: This survey conducted in Istanbul, the largest cosmopolitan city in Turkey with immigrants from all over the country, has a larger sample size than other previous studies, and therefore the reported prevalence rate of BD has a more acceptable confidence interval. This study aids in the estimation of the prevalence of BD in Turkey, and supports previous findings that Turkey has the highest prevalence rate of the disease in the world. PMID- 14521695 TI - Rare variants of Cutaneous Leishmaniasis: whitlow, paronychia, and sporotrichoid. AB - Cutaneous Leishmaniasis is endemic in certain areas of Pakistan, with the wet form of the disease being the most prevalent. It has a number of morphological variants, which are dependent on the immune status of the host, the subspecies of the Leishmania, and also, to some extent, on the site of involvement. We describe here a case of Leishmaniasis showing two very rare variants, whitlow and paronychial lesions, occurring concurrently with sporotrichoid spread. The patient responded to intramuscular sodium stibogluconate with resolution of the skin lesions. PMID- 14521696 TI - Enlarging pigmented patches on the hand. AB - This case illustrates the clinical and pathological features of Tinea nigra. Awareness of the characteristic features of this cutaneous infection, especially in areas where the organism is endemic, can ultimately lead clinicians to the appropriate diagnosis. If Tinea nigra should be considered in the differential diagnosis of palmoplantar pigmentation, potassium hydroxide (KOH) preparation may facilitate the diagnosis. PMID- 14521697 TI - Diagnostic performance and retention of acquired skills after dermatology elective. AB - BACKGROUND: The dermatology elective often constitutes the future physician's only exposure to dermatologic practice. Cost-benefit considerations dictate that the elective enables the students to acquire useful diagnostic expertise in the short time period available, and that this expertise is not rapidly forgotten after completion of the elective. METHODS: In order to determine the efficacy of a 2-week dermatology elective in promoting diagnostic skills and to ascertain the preservation of the acquired expertise, kodachrome slide-based multiple-choice examinations were administered to 84 sixth year medical students at the beginning (pre-test) and completion (post-test) of the elective. The long-term retention of diagnostic skills was evaluated by re-testing 4-9 months after completion of the elective (follow-up test). RESULTS: The mean scores of correct answers at the pre and post-test were 39.73% and 72.46%, respectively. The mean score at the follow up test for students with a previous dermatology elective was 80.22%, compared with a score of 46.00% for students who did not participate in the elective. CONCLUSIONS: The 2-week dermatology elective significantly improved the diagnostic skills of the participants, and the knowledge obtained during the elective was retained for at least 4-9 months. PMID- 14521698 TI - Cutaneous relapse of angioimmunoblastic lymphadenopathy-type peripheral T-cell lymphoma mimicking an exanthematous drug eruption. PMID- 14521699 TI - Bilateral pseudocyst of the auricle in a man with pruritus secondary to lymphoma. PMID- 14521700 TI - Disseminated cryptococcosis with cutaneous lesions complicating steroid therapy for Behcet's disease. PMID- 14521701 TI - Persistent generalized erythema in adult-onset Still's disease. PMID- 14521702 TI - Proteus syndrome with gingival hyperplasia. PMID- 14521703 TI - Comparison of trichloroacetic acid solution and cryosurgery in the treatment of solar lentigines. AB - BACKGROUND: Solar lentigines are benign keratinocytic proliferations resulting from prolonged and cumulative sun exposure. The newer photoselective lasers have become the mainstay of treatment. However, cryosurgery and trichloroacetic acid (TCA) solution are cost-effective alternatives in the treatment of solar lentigines. METHODS: Twenty-five patients with multiple solar lentigines over the dorsa of their hands were included. Each hand was treated randomly with either 30% TCA solution or liquid nitrogen spray. The efficacy of treatment was evaluated at 8 weeks. RESULTS: Cryosurgery was more likely to produce significant lightening of the lentigines than 30% TCA solution (P < 0.05) but was more painful and took longer to heal. CONCLUSION: Cryosurgery was found to be superior to TCA 30% solution in the treatment of solar lentigines. This study demonstrates that old-fashioned treatments for solar lentigines are still excellent and cost effective therapeutic choices. PMID- 14521704 TI - Treatment of acquired syndactyly by gauze-fixed epidermal graft after radiosurgery. AB - BACKGROUND: Acquired syndactyly is a rare disease that occurs mostly after disease, trauma, or other inflammatory conditions. It is usually treated by surgical incision with a flap or full-thickness skin graft, which is very invasive and requires hospitalization. OBJECTIVE: The objective was to treat acquired syndactyly with an epidermal graft by suction blister after radiosurgery, because this procedure is much less invasive and can be performed in an outpatient base. METHODS: A 65-year-old Korean woman had acquired syndactyly after various traditional treatments for tinea pedis. Five days after separation of syndactyly with radiosurgery, we prepared an epidermal sheet by suction blister, placed it on sterile meshed gauze, and applied it to the separated lesion. RESULTS: The patient's lesion was completely healed after 7 days. CONCLUSION: Gauze-fixed epidermal graft after radiosurgery is a very effective and simple treatment for shallow acquired syndactyly. PMID- 14521705 TI - Calcipotriol versus coal tar: a prospective randomized study in stable plaque psoriasis. AB - BACKGROUND: Topical therapies are the first line of treatment for patients with stable plaque psoriasis (SPP) affecting a limited body surface area. Very few trials comparing newer agents, such as 0.005% topical calcipotriol, with conventional modes of therapy, such as coal tar ointment, have been reported. METHODS: A prospective, right-left randomized, investigator-blinded study with a 12-week treatment period and an 8-week follow-up period was performed. Thirty-six patients with nearly bilaterally symmetrical SPP lesions on the limbs were instructed to apply 5% coal tar ointment overnight on one side once daily and 0.005% calcipotriol ointment on the other side twice a day. All patients were advised to expose both sides to the sun for 2 h every day. Psoriatic lesions and progress during treatment were evaluated using the severity (0-3) scale of erythema, scaling and induration (ESI score). Evaluation was carried out every 2 weeks during the treatment period and monthly during follow-up. At the end of 12 weeks, patients with > 75% reduction in the ESI score were considered to be markedly improved, those with 51-75% reduction to be moderately improved, those with 26-50% reduction to be minimally improved and those with < 25% to be non responders. Self-assessment by the patients regarding the efficacy and acceptability of the two modalities was on a five-point scale. Serum calcium, serum phosphate, total and differential serum proteins, 24-h urinary calcium and phosphate were monitored both at baseline and after completion of therapy. RESULTS: Thirty of the 36 recruited patients completed the study. The difference in clinical response between the two sides was statistically significant at 4, 6 and 8 weeks, with the percentage reduction in ESI score with calcipotriol being 65.7 +/- 12.2% compared with 45.8 +/- 16.6% with coal tar at 8 weeks (P < 0.01, t = 6.4). However, the difference in clinical response at 10 and 12 weeks between the two sides was not significant, with a mean reduction of 71.9 +/- 13.3% in ESI score on the calcipotriol-treated side compared with 69.4 +/- 15.4% with coal tar ointment (P > 0.05). In the follow-up period of 8 weeks, recurrence of lesions was noted in 10% of patients treated with calcipotriol compared with 16.7% in those treated with coal tar after an average period of 6 +/- 1.2 and 5 +/- 1.3 weeks, respectively (P > 0.05). CONCLUSIONS: It was found that 0.005% calcipotriol ointment produced a faster initial response and had better cosmetic acceptability in patients, although after a long period of treatment, i.e. 12 weeks, 5% coal tar ointment had comparable efficacy. There was no statistically significant difference in the relapse rates between the two modalities. PMID- 14521706 TI - Tinea corporis in human immunodeficiency virus-positive patients: case report and assessment of oral therapy. AB - Dermatophytosis in immunocompromised hosts is more varied and often more severe than in immunocompetent hosts. Early recognition and treatment with systemic therapy are important in human immunodeficiency virus (HIV)-positive patients in order to prevent severe infection. Potential drug resistance can occur due to chronic usage of systemic azole therapy in such patients, or the existence of atypical fungi. Although warnings have been made of possible drug interactions between certain antifungals and antiretroviral medications, only one combination has shown a clinically significant interaction. A case treated aggressively with oral terbinafine at the onset is presented. BACKGROUND: Dermatophytosis in immunocompromised hosts is more varied and often more severe than in immunocompetent hosts. Early recognition and treatment with systemic therapy are important in human immunodeficiency virus (HIV)-positive patients in order to prevent severe infection. OBJECTIVE: To analyze potential therapies for dermatophyte infections in immunocompromised patients and risk of drug resistance and interactions with antiretroviral medications. METHODS: Literature search based on MEDLINE (1966-March 2003) and additional references obtained from cross referencing retrieved articles. All information deemed relevant by the reviewers was included. A case study was employed to exemplify the usage of this information in patient care. RESULTS: Although warnings have been made of possible drug interactions between certain antifungals and antiretroviral medications, only one combination has shown a clinically significant interaction. CONCLUSIONS: When considering drug interactions and side-effects, there are no clinically significant reasons to avoid any oral antifungal for dermatophytosis in the HIV-positive patient. PMID- 14521707 TI - Kaposi's sarcoma is not a neoplasm let alone a sarcoma. PMID- 14521708 TI - Global Solar UV Index: a physician's tool for fighting the skin cancer epidemic. PMID- 14521709 TI - Cholinergic urticaria successfully treated with scopolamine butylbromide. PMID- 14521710 TI - The Maria M. Duran Committee and Award of the International Society of Dermatology. PMID- 14521713 TI - Geographic information systems and the spiritual dimension of health: a short position paper. AB - The WHO's well known definition of health stressed the indivisibility of human well-being, physical and otherwise, by stating that health is "not merely the absence of disease or infirmity". The spiritual dimension of health is well covered in the medical literature. Different locations on Earth are associated with different interrelated profiles: physical, biological, environmental, socio economic, cultural, and also spiritual profiles, that do affect and are affected by health (including its spiritual dimension), disease, healthcare, and pastoral care. A number of reviews have been recently published covering the use of Geographic Information Systems (GIS) in understanding and harnessing the importance of location in the health sector. However, no publication so far has discussed the role of GIS in relation to the spiritual dimension of health. This position paper is an attempt to fill in this gap without going into deep details. GIS role in pastoral care ranges from assisting in pattern and trend detection, and in informed decision-making and resource management, to providing routing and educational functions, and even assessing the impact of missionary radio broadcasts. A review of some of the software tools that are currently available in this field is also provided. GIS are ideal tools for improving and coordinating the integration of the health (physical), social, and spiritual/ pastoral dimensions of individual and community care. However to achieve the full potential of GIS in these areas, we still need to combat many cultural and organisational barriers, while making the tools cheaper and much easier to learn and use. PMID- 14521712 TI - A common polymorphism in the oxygen-dependent degradation (ODD) domain of hypoxia inducible factor-1alpha (HIF-1alpha) does not impair Pro-564 hydroxylation. AB - BACKGROUND: The hypoxia-inducible factor (HIF) transcription complex, which is activated by low oxygen tension, controls a diverse range of cellular processes including angiogenesis and erythropoiesis. Under normoxic conditions, the alpha subunit of HIF is rapidly degraded in a manner dependent on hydroxylation of two conserved proline residues at positions 402 and 564 in HIF-1alpha in the oxygen dependent degradation (ODD) domain. This allows subsequent recognition by the von Hippel-Lindau (VHL) tumor suppressor protein, which targets HIF for degradation by the ubiquitin-proteasome pathway. Under hypoxic conditions, prolyl hydroxylation of HIF is inhibited, allowing it to escape VHL-mediated degradation. The transcriptional regulation of the erythropoietin gene by HIF raises the possibility that HIF may play a role in disorders of erythropoiesis, such as idiopathic erythrocytosis (IE). RESULTS: Patients with IE were screened for changes in the HIF-1alpha coding sequence, and a change in the ODD domain that converts Pro-582 to Ser was identified in several patients. This same change, however, was also detected at a significant frequency, 0.073, in unaffected controls compared to 0.109 in the IE patient group. In vitro hydroxylation assays examining this amino acid change failed to reveal a discernible effect on HIF hydroxylation at Pro-564. CONCLUSION: The Pro582Ser change represents a common polymorphism of HIF-1alpha that does not impair HIF 1alpha prolyl hydroxylation. Although the Pro582Ser polymorphism is located in the ODD domain of HIF-1alpha it does not diminish the association of HIF-1alpha with VHL. Thus, it is unlikely that this polymorphism accounts for the erythrocytosis in the group of IE patients studied. PMID- 14521714 TI - Gamma (gamma) tocopherol upregulates peroxisome proliferator activated receptor (PPAR) gamma (gamma) expression in SW 480 human colon cancer cell lines. AB - BACKGROUND: Tocopherols are lipid soluble antioxidants that exist as eight structurally different isoforms. The intake of gamma-tocopherol is higher than alpha-tocopherol in the average US diet. The clinical results of the effects of vitamin E as a cancer preventive agent have been inconsistent. All published clinical trials with vitamin E have used alpha-tocopherol. Recent epidemiological, experimental and molecular studies suggest that gamma-tocopherol may be a more potent chemopreventive form of vitamin E compared to the more studied alpha-tocopherol. Gamma-tocopherol exhibits differences in its ability to detoxify nitrogen dioxide, growth inhibitory effects on selected cancer cell lines, inhibition of neoplastic transformation in embryonic fibroblasts, and inhibition of cyclooxygenase-2 (COX-2) activity in macrophages and epithelial cells. Peroxisome proliferator activator receptor gamma (PPARgamma) is a promising molecular target for colon cancer prevention. Upregulation of PPARgamma activity is anticarcinogenic through its effects on downstream genes that affect cellular proliferation and apoptosis. The thiazolidine class of drugs are powerful PPARgamma ligands. Vitamin E has structural similarity to the thiazolidine, troglitazone. In this investigation, we tested the effects of both alpha and gamma tocopherol on the expression of PPARgamma mRNA and protein in SW 480 colon cancer cell lines. We also measured the intracellular concentrations of vitamin E in SW 480 colon cancer cell lines. RESULTS: We have discovered that the alpha and gamma isoforms of vitamin E upregulate PPARgamma mRNA and protein expression in the SW480 colon cancer cell lines. gamma-Tocopherol is a better modulator of PPARgamma expression than alpha-tocopherol at the concentrations tested. Intracellular concentrations increased as the vitamin E concentration added to the media was increased. Further, gamma-tocopherol-treated cells have higher intracellular tocopherol concentrations than those treated with the same concentrations of alpha-tocopherol. CONCLUSION: Our data suggest that both alpha and gamma tocopherol can upregulate the expression of PPARgamma which is considered an important molecular target for colon cancer chemoprevention. We show that the expression of PPARgamma mRNA and protein are increased and these effects are more pronounced with gamma-tocopherol. Gamma-tocopherol's ability to upregulate PPARgamma expression and achieve higher intracellular concentrations in the colonic tissue may be relevant to colon cancer prevention. We also show that the intracellular concentrations of gamma-tocopherol are several fold higher than alpha-tocopherol. Further work on other colon cancer cell lines are required to quantitate differences in the ability of these forms of vitamin E to induce apoptosis, suppress cell proliferation and act as PPAR ligands as well as determine their effects in conjunction with other chemopreventive agents. Upregulation of PPARgamma by the tocopherols and in particular by gamma tocopherol may have relevance not only to cancer prevention but also to the management of inflammatory and cardiovascular disorders. PMID- 14521716 TI - Penile metastasis of prostatic adenocarcinoma: Report of two cases and review of literature. AB - BACKGROUND: Carcinoma of the prostate metastasising to the penis is rare. These patients have a poor prognosis receiving various treatment modalities. CASE PRESENTATION: Two such patients are discussed here having received differing therapeutic regimes, pointing out the necessity for standardised palliative treatment rather than radical therapy. CONCLUSION: Management of patients with penile metastases from carcinoma of the prostate should emphasise improving quality of life with palliative treatment rather than radical therapy PMID- 14521715 TI - PLZF is a negative regulator of retinoic acid receptor transcriptional activity. AB - BACKGROUND: Retinoic acid receptors (RARs) are ligand-regulated transcription factors controlling cellular proliferation and differentiation. Receptor interacting proteins such as corepressors and coactivators play a crucial role in specifying the overall transcriptional activity of the receptor in response to ligand treatment. Little is known however on how receptor activity is controlled by intermediary factors which interact with RARs in a ligand-independent manner. RESULTS: We have identified the promyelocytic leukemia zinc finger protein (PLZF), a transcriptional corepressor, to be a RAR-interacting protein using the yeast two-hybrid assay. We confirmed this interaction by GST-pull down assays and show that the PLZF N-terminal zinc finger domain is necessary and sufficient for PLZF to bind RAR. The RAR ligand binding domain displayed the highest affinity for PLZF, but corepressor and coactivator binding interfaces did not contribute to PLZF recruitment. The interaction was ligand-independent and correlated to a decreased transcriptional activity of the RXR-RAR heterodimer upon overexpression of PLZF. A similar transcriptional interference could be observed with the estrogen receptor alpha and the glucocorticoid receptor. We further show that PLZF is likely to act by preventing RXR-RAR heterodimerization, both in-vitro and in intact cells. CONCLUSION: Thus RAR and PLZF interact physically and functionally. Intriguingly, these two transcription factors play a determining role in hematopoiesis and regionalization of the hindbrain and may, upon chromosomal translocation, form fusion proteins. Our observations therefore define a novel mechanism by which RARs activity may be controlled. PMID- 14521717 TI - Impact of nausea and vomiting on quality of life in cancer patients during chemotherapy. AB - It is commonly claimed that the nausea and vomiting accompanying cytotoxic chemotherapy have a negative impact on health-related quality of life. While this may seem self-evident, until a few years ago there was little empirical data demonstrating that the failure to control postchemotherapy emesis affects aspects of quality of life. In spite of their limitations, several observational studies showed that nausea and vomiting associated with chemotherapy induced a decrease in health-related quality of life with respect to patients without nausea and vomiting. This has also been demonstrated after the adjustment for health-related quality of life before chemotherapy that is an important prognostic factor of chemotherapy-induced nausea and vomiting. Furthermore, one study suggests that the optimal time of assessment of quality of life to evaluate the impact of chemotherapy-induced nausea and vomiting is day 4 if a 3-day recall period is used or day 8 when the recall period is 7 days. In double-blind studies the efficacy, tolerability and impact on quality of life of the 5-HT3 receptor antagonists was superior with respect to metoclopramide, alizapride and prochlorperazine. Similar results have been achieved with the combination of ondansetron with dexamethasone, the standard treatment for the prevention of acute emesis induced by moderately emetogenic chemotherapy, with respect to the metoclopramide plus dexamethasone combination. Instead, in another double-blind study, in patients submitted to moderately emetogenic chemotherapy, a 5-HT3 antagonist did not seem to significantly increase complete protection from delayed emesis and the patients' quality of life with respect to dexamethasone alone. In conclusion, the evaluation of quality of life in randomized trials comparing different antiemetic drugs for the prevention of chemotherapy-induced nausea and vomiting can add important information useful for the choice of the optimal antiemetic treatment. PMID- 14521718 TI - The Women's Health Questionnaire (WHQ): Frequently Asked Questions (FAQ). AB - The Women's Health Questionnaire (WHQ) is a measure of mid-aged women's emotional and physical health. Since its publication in 1992 the WHQ has been widely used in multinational clinical trials, in epidemiological studies as well as in the evaluation of non-medical treatments. In particular the WHQ has been included as a quality of life measure in trials of hormonal preparations for peri and post menopausal women and in studies using a variety of preventative interventions for mid-aged and older women. The questionnaire was developed in English and standardised on a sample of women aged 45-65 years. It is reliable, has good concurrent validity and is sensitive to detecting change, and is available in 27 languages. The range of subscales included in the WHQ enable a detailed assessment of dimensions of emotional and physical health, such as depression, anxiety, sleep problems, somatic symptoms, with optional subscales for menstrual problems and sexual difficulties. The WHQ is the first measure to be included in the MAPI Research Institute's database, the International Health-related Quality of Life Outcomes Database (IQOD). Drawing upon data from international studies this project aims to produce reference values for cross-culturally valid, reliable and responsive quality of life instruments. In addition to this work, a revised shorter version of the WHQ is currently being developed. PMID- 14521720 TI - Application of magnetic techniques in the field of drug discovery and biomedicine. AB - Magnetic separation technology, using magnetic particles, is quick and easy method for sensitive and reliable capture of specific proteins, genetic material and other biomolecules. The technique offers an advantage in terms of subjecting the analyte to very little mechanical stress compared to other methods. Secondly, these methods are non-laborious, cheap and often highly scalable. Moreover, techniques employing magnetism are more amenable to automation and miniaturization. Now that the human genome is sequenced and about 30,000 genes are annotated, the next step is to identify the function of these individual genes, carrying out genotyping studies for allelic variation and SNP analysis, ultimately leading to identification of novel drug targets. In this post-genomic era, technologies based on magnetic separation are becoming an integral part of todays biology laboratory. This article briefly reviews the selected applications of magnetic separation techniques in the field of biotechnology, biomedicine and drug discovery. PMID- 14521719 TI - A review of mothers' prenatal and postnatal quality of life. AB - BACKGROUND: Contemporary broad descriptions of health and well-being are reflected in an increasing appreciation of quality of life issues; in turn this has led to a growing number of tools to measure this. METHODS: This paper reviews articles cited in MEDLINE, CINAHL and BIDS which have addressed the concept of quality of life in pregnancy and the period following childbirth. RESULTS: It describes five groups of articles: those explicitly assessing quality of life in this area; those using broader health assessments as an indicator of quality of life; those articles equating quality of life with certain pregnancy outcomes in identified groups of patients; those studies which identify the possibility of pregnancy as an outcome measure and infer from this that quality of life has been improved; and those articles which are themselves reviews or commentaries of pregnancy and childbirth and which identify quality of life as a feature. CONCLUSIONS: The term 'quality of life' is used inconsistently in the literature. There are few quality of life tools specifically designed for the maternity care setting. Improved or adversely affected quality of life is frequently inferred from certain clinical conditions. PMID- 14521722 TI - [The relationship between insulin resistance and endothelium-dependent vasodilatation in obese subjects]. AB - OBJECTIVE: To study the relationship between insulin resistance and endothelium dependent vasodilatation function in obese subjects with normal blood glucose, serum cholesterol and blood pressure and to explore the non-traditional risk factors associated with atherosclerosis. METHODS: 24 euglycemic obese males (OB) and 12 age and sex matched lean controls (Lean) underwent oral glucose tolerance test (OGTT), euglycemic hyperinsulinemia clamp study with an insulin infusion rate of 120 mU/m(2)/min to evaluate the peripheral glucose disposal rate (GDR) in steady-state, brachial artery ultrasound study to assess the endothelium dependent vasodilation (EDV) and independent vasodilation (EIV), the combined intima-media thickness (IMT) of common carotid arteries examined by high resolution ultrasound, and fasting lipids profile and free fatty acids (FFAs) concentration. RESULTS: The blood glucose and serum cholesterol levels were all normal in the OB group but the serum FFA and triglyceride concentrations were elevated compared with those of the controls (FFA: 888 microM/l +/- 158 microM/l vs 508 microM/l +/- 137 microM/l, P < 0.05; TG: 1.8 mM/l +/- 0.6 mM/l vs 1.1 mM/l +/- 0.5 mM/l, P < 0.05) The GDR in steady-state was 6.7 mg x min(-1) x kg(-1) +/- 1.4 mg x min(-1) x kg(-1) in the OB group, significantly lower than that of the controls (12.2 mg x min(-1) x kg(-1) +/- 3.1 mg x min(-1) x kg(-1), P < 0.05). In the OB group the flow-mediated dilation of the brachial artery caused by reactive hyperemia (EDV) were impaired and the relaxation time was shorter than that of the controls (P < 0.05) but the EIV was not significantly different between these two groups. There was a significantly positive correlation between GDR and EDV (r = 0.438, P < 0.05). The IMT of common carotid arteries was thicker in the OB group than in the control group (0.50 mm +/- 0.02 mm vs 0.34 mm +/- 0.19 mm). CONCLUSION: The endothelium-dependent vasodilation was impaired in obese subjects even though without hyperglycemia, hypercholesterolemia, and hypertension. Traditional cardiovascular risk factors cannot fully account for the increased risk of cardiovascular disease in metabolic syndrome. Elevated circulating FFA concentration, is characteristic of metabolic syndrome, may play a role in endothelial dysfunction. PMID- 14521721 TI - Effects of STI571 (gleevec) on pancreatic cancer cell growth. AB - BACKGROUND: Pancreatic cancer is an aggressive malignancy characterized by low responsiveness to chemotherapy and radiotherapy. This resistance is partly due to the overexpression of several tyrosine kinase receptors and their ligands. STI571 has specific activity in inhibiting c-kit, PDGF and Abl receptor tyrosine kinases and has proven successful in the treatment of CML and GIST patients. Here, we investigated the potential role of STI571 in pancreatic cancer. RESULTS: The GI50 of STI571 as well as the effects of STI571 on growth factor actions in pancreatic cell lines were analyzed using the MTT assay. FACS analysis using Annexin and PI staining was performed to study cell cycle, apoptosis, and cell death. Western blot analysis was carried out to investigate MAP kinase and receptor tyrosine kinase phosphorylation. STI571 inhibited cell proliferation in pancreatic cancer cell lines with GI50 concentrations ranging from 17 to 31.5 microM. EGF, IGF-1, and FGF-2 but not PDGF exerted growth stimulatory effects in pancreatic cancer cell lines. STI571 only partly blocked these effects on cell growth, and did not abrogate growth factor-induced receptor and MAPK phosphorylation. CONCLUSION: Our data demonstrate that STI571 inhibits pancreatic cancer cell growth with high GI50 concentrations through tyrosine-kinase receptor independent pathways. The clinical application of STI571 in pancreatic cancer is therefore rather doubtful. PMID- 14521724 TI - [Influence of different procedures of alimentary tract reconstruction after total gastrectomy for gastric cancer on the nutrition and metabolism of patients: a prospective clinical study]. AB - OBJECTIVE: To investigate the influence of different alimentary tract reconstruction procedures after total gastrectomy for treatment of gastric cancer on nutrition and metabolism and explore an ideal reconstruction procedure. METHODS: A total of 149 patients with gastric cancer who had undergone total gastrectomy were randomly allocated into 5 groups of 30 patients (except the group HLD with 29 cases) to undergo 5 different alimentary tract reconstruction procedures: simple esophagojejunostomy using Roux-en-Y technique (RY), P pouch with Roux-en-Y reconstruction (PRY), jejunal pouch reconstruction according to Hunt-Lawrence technique (HL), jejunal pouch original interposition reconstruction modified by the authors (JOP), and Hunt-Lawrence reconstruction technique maintaining duodenal passage (HLD). Three and six months after operation, quality of life (Visick grade), PNI; body weight; and serum nutritional parameters, including albumin, (ALB), total protein (TP), transferrin (TF), hemoglobin (HB), and serum iron (SI), were evaluated. RESULTS: In comparison with those of the PRY, HL, JOP, and HLD groups, the patients of the RY group show greater weigh loss, and lower ALB, TP, and TF (all P < 0.05). The HB, SI, and TS levels in the JOP group and HLD group were significantly higher than those in the RY, PRY, and HL groups (all P < 0.05). CONCLUSION: Different procedures of alimentary tract reconstruction after total gastrectomy have great influence on the patients' nutrition at different degrees. The patients undergoing the procedures with a reservoir show higher serum nutritional parameters and better body weight. The volume of reservoir has no major clinical importance. The jejunal pouch original interposition reconstruction modified by the authors (JOP), constructing a gastric reservoir and maintaining the alimentary tract flowing through the duodenum is an ideal choice for the reconstruction after total gastrectomy. PMID- 14521723 TI - [Reduced resistin levels in patients with type 2 diabetes mellitus]. AB - OBJECTIVE: To measure the serum resistin level of patients with type 2 diabetes mellitus so as to examine whether there exists a relationship between resistin, obesity and diabetes. METHODS: ELISA was used to examine the fasting serum resistin, leptin, and true insulin and those 2-hours after taking 75-g glucose in 51 untreated type 2 diabetic patients, 30 males and 21 females, and 52 sex and age-matched normal control subjects. Blood glucose, blood pressure, height, weight, waist circumstance, hip girth were measured. Body mass index (BMI), waist to hip ratio (WHR), and quantitative insulin sensitivity check index (QUICKI) were calculated. RESULTS: In comparison with the control, the diabetic group had higher waist-to-hip ratio (WHR) and serum insulin levels (P < 0.05), but significantly lower resistin levels both in the fasting status (23 ng/ml +/- 15 ng/ml vs 30 ng/ml +/- 18 ng/ml, P < 0.05) and 2 hours after glucose loading (22 ng/ml +/- 11 ng/ml vs 31 ng/ml +/- 15 ng/ml, P < 0.001). The leptin level was not statistically different between the two groups (P > 0.05). The resistin level 2 hours after glucose loading was not significantly different between these 2 groups. Correlation analysis demonstrated that fasting resistin level was not correlated with sex, BMI, leptin, and blood pressure, but positively correlated with QUICKI (r = 0.30, P < 0.01) and negatively correlated with blood glucose (r = -0.21, P < 0.05). CONCLUSION: The serum resistin level of patients with type 2 diabetes is reduced rather than increased in fasting status and 2 hours after glucose taking. Resistin may not be the major link between obesity and diabetes in human beings. Since human resistin level is positively correlated with insulin sensitivity, the use of term "resistin", originally for its resistance to insulin, may be somewhat premature. PMID- 14521725 TI - [Correlation between polymorphisms of interleukin-1beta and RN genes and risk of gastric carcinoma: a case-control study]. AB - OBJECTIVE: To investigate the correlation between polymorphisms of interleukin 1beta and RN genes and risk of gastric carcinoma. METHODS: PCR and denaturing high-performance liquid chromatography (DHPLC) were used to analyze the IL-1beta 31 and -511 C/T polymorphisms and to genotype the IL-1RN penta-allelic variable number of tandem repeats (VNTR): of the DNA from the peripheral blood of 143 patients of gastric carcinoma, 97 from Linqu County, Shandong Province, and 46 cases from the specimen bank of Beijing Cancer Hospital, and 337 controls without gastric carcinoma from Linqu County, most of which suffered from other gastric diseases. RESULTS: IL-1-511 polymorphisms (carriers of IL-1beta-511T) were correlated with the increased risk of interstitial gastric carcinoma in both sexes (OR = 3.833, 95% CI: 2.282 approximately 6.439) and increased risk of diffuse gastric carcinoma in males (OR = 3.464, 95% CI: 1.394 approximately 8.608). The frequencies of IL-1beta-31 polymorphisms and IL-1IRN VNTR were not different between the gastric carcinoma group and the control group, IL-1beta-511 T carrier could be seen in the controls at different precancerous stages, for chronic atrophic gastritis (OR = 5.164, 95% CI: 2.661 approximately 10022), for interstitial metaplasia (OR = 5.093, 95% CI: 2.708 approximately 9.463), and for dysphasia (OR = 8.438, 95% CI: 3.939 approximately 18.073). CONCLUSION: IL-1beta 31 genotypes and. IL-2RN VNTR do not increase the risk of gastric carcinoma. The IL-1beta-511 T genotype increases the risk of gastric carcinoma in every precancerous stage. PMID- 14521726 TI - [Frequent 4 977 bp deletion of mitochondrial DNA in tumor cell lines, solid tumors and precancerous lesions of human stomach]. AB - OBJECTIVE: To clarify the frequency of mtDNA 4 977 bp deletion in tumor cell lines, solid tumors, patient's serum of gastric tumor and to find a easy and exact method to diagnose gastric tumor. METHODS: Primer-shift PCR method was used in 13 gastric tumor cell lines, 52 cases of gastric fresh tumor tissues matched the adjacent normal tissues, 40 cases of patient's serum of gastric tumor and 40 cases of normal serums for analysis of mtDNA deletion. RESULTS: Frequency of 4 977 bp mtDNA deletion was detected in 12 of 13 (92.3%) tumor cell lines, 38 of 52 (73.1%) cases of solid tumor tissues, 27 of 52 (52%) adjacent normal tissues, 17 of 40 (42.5%) patient's serum and 8 of 40 (20%) normal serums. Further more, in 2 of 10 pairs microdissected specimens, we found this deletion occurred not only in primary tumor but also in intestinal metaplasia comparing with no deletion was found in normal tissues. The frequency of this deletion was statistically significantly higher in the gastric tumor tissues and serums than in the adjacent normal tissues and serums. A good correlation between the deletion and young age of patients was representative in our data. CONCLUSION: mtDNA 4 977 bp deletion maybe play an important role in the carcinogenesis of human gastric mucous and maybe has happened in the tumor cell malignant transformation. To detect this deletion in patient's serum is a easy and exact method and maybe become a potential tumor marker. PMID- 14521727 TI - [Transmaxillary approach for micro resection of clivus tumors, clinical analysis of 16 cases]. AB - OBJECTIVE: To investigate the indications, key point, advantages, and disadvantages of transmaxillary approach for microsurgical removal of clivus tumors. METHODS: The clinical data of 16 consecutive patients with clivus tumors operated upon through transmaxillary approach, based on Le Fort I ostectomy, July 1999 to May 2002 were retrospectively reviewed. RESULTS: Of the 16 patients, 8 patients suffered from chordoma, 2 from angiofibroma, 1 from pituitary adenoma, 1 from chondroma, 1 from adenocarcinoma of grade II, 1 from mucoid cyst, 1 from myeloma, and 1 from fibrous dysplasia of bone. Four out of the 16 cases were with recurrent tumors after operation performed in other hospitals. The operation field of all 16 patients was satisfactorily exposed. The tumors of 10 cases were totally removed under microscope, and 6 tumors were subtotally removed. The complications included rhinorrhea in 1 case and malocclusion in 3 cases. There was no operative mortality. All patients were followed up for 6 approximately 40 months (ion average 20 months) with MRI. Four of them had local recurrence within 1 year, 1.5 years, 2 years, and 9 months respectively after operational and have lived with tumors till now. The remainders had resumed their normal life. CONCLUSION: An ideal approach, transmaxillary approach facilitates the resection of clivus tumors, with satisfying exposure and fewer complications. PMID- 14521728 TI - [Expression and recombination mechanism of SNC73 (IgHalpha1) in human epithelial cancer cell line]. AB - OBJECTIVE: To study if the gene SNC73 (IgHalpha1) is expressed in human epithelial cancer cell line and to interpret the recombination mechanism. METHODS: Human epithelial cancer cells of SW480 line were cultured. RT-PCR and Western blotting were used to examine the expression of SNC73, recombination activating gene 1 (RAG1), and RAG2. The RT-PCR products were confirmed by sequencing. Immunohistochemistry was used to detect the expression of IgHalpha1, Igkappa, and Iglambda in these epithelial cancer cells. RESULTS: The human epithelial cancer cell line (SW480) positively expressed SNC73, RAG1, and RAG2. IgHalpha1 and Igkappa was strongly expressed in SW480 cells, but Iglambda was undetectable. The sequence of the constant region of SNC73 in SW480 cells is identical to that of IgA1. Both sequencing and Western blotting showed that the RAG1 and RAG2 expressed in SW480 cells were identical to that expressed in pre-B lymphocytes. CONCLUSION: Immunoglobulin alpha-1 gene is expressed in non-lymphoid cells, which may be a potential genetic marker for the development of colorectal cancer. Recombination signal sequence (RSS)-mediated recombination may take part in the rearrangement of immunoglobulin alpha-1 gene in human epithelial cancer cell line. PMID- 14521729 TI - [The effect of activated nuclear factor-kappaB in pathogenesis of acute pancreatitis]. AB - OBJECTIVE: To explore the effect of the activated NF-kappaB and the interaction between activated NF-kappaB and tumor necrosis factor (TNF)-alpha in the process of acute pancreatitis. METHODS: 64 Wistar rats were randomly divided into two groups of 32 rats: pancreatitis group (to be made pancreatitis models) and control group. 1.5, 3.0, 6.0, and 12.0 hours after the onset of experiment the amount of ascitic fluid was measured, blood was extracted from abdominal aorta, changes of pancreas was observed, pancreatic tissues were stained with HE, and flow cytometry (FCM) and enzyme linked immuno-sorbent assay (ELISA) were used to examine the content of TNF-alpha protein and activation of NF-kappaB (number of positive cells/50 micro l) in the pancreatic tissues. The contents of amylase and lipase in plasma were examined. The pathology of pancreatic tissue was graded. RESULTS: The levels of activated NF-kappaB and TNF-alpha protein in the pancreatic tissue were all significantly higher than those of the control group at any time point (all P < 0.05). The levels of activation of NF-kappaB of the pancreatitis group at any time point were significantly higher than those of the control group (all P < 0.01) and reached its maximum about 3.0 hours after the onset of experiment and then declined. The levels of plasma amylase and lipase were significantly higher in the pancreatic group than in the control group at any rime point. (all P < 0.01). In the pancreatic group severe edema and congestion were found at the 3.0 h time point and bleeding and necrosis were found at the 6.0-hour time point. CONCLUSION: Activated NF-kappaB as initial factor plays an important role in the pathogenesis of acute pancreatitis, activates a lot of inflammatory media, and induces cascading reaction of inflammation. TNF-alpha is a pivotal factor in pancreatitis pathogenesis, it cooperates with amylase and lipase to intensify the leisure in pancreatic tissue resulting bleeding and necrosis. PMID- 14521730 TI - [Inhibition of angiogenesis in pancreatic carcinoma by cyclooxygenase-2 antisense oligodeoxynucleotides]. AB - OBJECTIVE: To investigate the effect of cyclooxygenase-2 antisense oligodeoxynucleotides (COX-2 AS-ODNs) on the angiogenesis in pancreatic carcinoma and to evaluate the intermediary effect of prostaglandin 2 in this process. METHODS: Specific targeting COX-2 AS-ODNs were designed and synthesized, and transfected into the PC3 human pancreatic carcinoma cells cultured in vitro. Fluorescence microscopy was used to observe the PC3 cells 0.12. 24, 40, and 72 hours after the transfection. the second cultured PC3 cells were divided into 5 groups: control group, Lipo group (transfected with Lipofectin only), C1 group (transfected with 1 micro g COX-2 AS-ODN + Lipo/well), C2 group (transfected with 2 micro g COX-2 AS-ODN + Lipo/well), and C3 group (transfected with 3 micro g COX 2 AS-ODN + Lipo/well). RT-PCR was used to observe the expression of COX-2 mRNA in the PC3 cells. The third batch of PC3 cells were transfected with 3 micro g COX-2 AS-ODN + Lipo/well, and the expression of COX-2 mRNA was observed 0, 12, 24, 48, and 72 hours later by RT-PCR. 3 micro g COX-2 AS-ODN + Lipo/bottle and 9 micro g COX-2 AS-ODN + Lipo/bottle were added into the cultured PC3 cells and Western blotting was used to observe the expression of COX-2 protein 24 hours later. 24 chicken eggs were inoculated with PC3 cells into the chorio-allantoic membrane and then divided equally into 5 groups; control group, Lipo group, COX-2 AS-ODN + Lipo group, and COX-2 AS-ODN + Lipo + PGE2 group. Leica microscopy was used to observe the angiogenesis in the transplanted carcinoma. RESULTS: RT-PCR showed that the downregulation of expression of COX-2 mRNA in the PC3 cells with the increase of the COX-2 AS-ODN concentration, peaking at the concentration of 0.2 micro mol/L. The effect of COX-2 AS-ODN was strongest by the 12th hour after transfection and then began to decrease and basically disappeared 48 hours after. Western blotting showed that COX-2 AS-ODN, especially that of the concentration of the expression of 9 micro g/bottle, inhibited the expression of COX-2 AS-ODN. The angiogenesis of the transplanted carcinoma in the eggs was significantly inhibited in the Lipo + COX-2 AS-ODN group, the density of newly generated vessels in the Lipo + COX-2 AS-ODN + PGE2 group was between those of the other 2 groups. CONCLUSION: COX-2 AS-ODN significantly inhibits the angiogenesis in the pancreatic carcinoma. Endogenous COX-2 AS-ODN may play an important role in such a process and PGE2 may play an intermediate role therein. PMID- 14521731 TI - [The inhibitory effect of hyporesponsive peptide on human leukocyte antigen DRbeta1 specific T cell activation]. AB - OBJECTIVE: To evaluate the inhibitory effect of human leukocyte antigen (HLA) DRbeta1 specific collagen II (CII) peptides with substitutions of TCR binding residues on T cell activation, and explore a new therapeutic strategy for T cell mediated autoimmune diseases by interfering with antigen recognition of T Cell receptor (TCR). METHODS: Non-TCR binding peptides were designed by computer modeling based on interaction of HLA DR1. The modified CII263-272. Intracellular transfer of the modified CII peptide and its binding to HLA DR1 were studied using confocal microscopy and fluorescence-activated cell sorter (FACS). The effects of altered peptides on T cell activation were evaluated using an antigen presenting system consisting of HLA-DR1 transgenic APC and CII specific T cells. RESULTS: Computer modeling showed the side chains of 263 (F), 266 (E) fit in the peptide binding groove, and form hydrogen bond with alpha1, beta1 chain of HLA DR1. The side chains of TCR specific 267 (Q) and 270 (K) protruded out of the groove, which might be TCR recognizing residues. The modified CII peptides with intact HLA-DR1 binding residues were bound to intracellular HLA-DR1 and expressed on cell surface. The modified peptides with single residue substitution of 267 270 and consecutive substitution of 268-270 showed a hyporesponsive T cell activation. Altered peptides 270A, sub268-270 could significantly suppress the T cell activation induced by CII263-272. CONCLUSION: The altered peptides with substitution of TCR binding residues are hyporesponsive in T cell activation, and may competitively inhibit the T cell activation in T cell mediated autoimmune diseases. PMID- 14521732 TI - [The correlation between the electroencephalogram findings and clinical manifestations of Creutzfeldt-Jakob disease]. AB - OBJECTIVE: To investigate the correlation between the EEG findings and the clinical signs of Creutzfeldt-Jakob disease (CJD). METHODS: Serial standard recording during different stage of disease was conducted among 13 patients with CJD, 10 males and 3 females, with the year of onset of 58.2 (49 approximately 65). The EEG findings were analyzed in combination of the clinical symptoms and signs. RESULTS: 12 of the 13 patients showed typical periodic sharp wave complexes (PSWCs), occurring after the onset of myoclonus and akinetic mutism in 8 patients, both after myoconus and before akinetic mutism in 2 cases, and just at the onset of akinetic mutism before or after the onset of myoclonus respectively in 2 cases. Different EEG features were observed in different stages of disease. Frontal intermittent rhythmic delta activity (FIRDA) and frontal intermittent rhythmic triphasic slow wave, which was replaced by PSWC later, were seen in 4 cases. CONCLUSION: PSWC has a close association with myoclonus and akinetic mutism. The best time of EEG recording is when myoclonus and/or akinetic mutism happen. FIRDA and frontal intermittent rhythmic triphasic slow wave may be hint of the onset of PSWC. PMID- 14521733 TI - [Adeno-associated virus vector carrying human minidystrophin gene SMCKA3999 effectively ameliorates dystrophic pathology in mdx model mice]. AB - OBJECTIVE: Duchenne muscular dystrophy (DMD) is the most common and letal genetic skeletal muscle disorder, caused by recessive mutations in the dystrophin gene and no treatment is available. The present paper is aimed to study if recombinant adeno-associated virus vector (rAAV) mediated dystrophin minigene SMCKA3999 could effectively ameliorates dystrophic pathology in mdx model mice. METHODS: We used dystrophin minigene SMCKA3999 to construct rAAVSMCKA3999. When injected into the skeletal muscle of mdx mice (DMD model), we adopted methods of immunofluorescent (IF) staining, Evans Blue and electromicroscopy to observe if rAAVSMCKA3999 could effectively ameliorates dystrophic pathology in mdx model mice. RESULTS: rAAVSMCK3999 resulted in efficient and stable expression and restoration of the missing dystrophin onto the plasma membrane. rAAVSMCKA3999 can also protect myofiber membrane integrity. For the first time we prove that rAAVSMCKA3999 can improve ultrastructure changes of DMD. CONCLUSION: We have demonstrated the effectiveness of rAAVSMCKA3999 in correcting pathology changes of skeletal muscle, which offer a promising avenue for DMD gene therapy. PMID- 14521735 TI - [Plasmid pUDKH gene therapy of rat acute hindlimb ischemia: an experimental study]. AB - OBJECTIVE: To investigate the effect of plasmid pUDKH carrying human hepatocyte growth factor (HGF) gene on rat acute limb ischemia and to find the lowest effective dosage. METHODS: Ligation of femoral artery of one hindlimb in Wistar rats was performed. The rats were randomly divided into 5 groups of 10 rats: pUDKH 50 microg, 100 microg, 200 microg, and 400 microg groups and a vacant vector pUDK group (control group). The plasmids were injected once directly into the ischemic limb muscle (5 sites around ligation position) immediately after ligation. At day 10, the muscles were removed and stained with H.E. to assess histologically the blood vessel formation. HGF expression was detected in the muscle tissue of another 12 rats on days 3, 5, 10, 14, 21, 30 after injection of pUDKH or pUDK, 200 microg each per animal. RESULTS: HGF expression was detected clearly in muscle tissue on days 3, 5, 10, 14 in pUDKH groups. In HGF-transfected animals (except for 50 microg group) neoformation of blood vessels in muscles and soft tissues was found, while it was not seen in controls. By semi-quantitation of the degree of vessel formation, significant differences between pUDKH groups (0.96 +/- 0.07, 1.97 +/- 0.91, 2.26 +/- 1.00 for 100 microg, 200 microg, 400 microg, respectively) and the control group (0.27 +/- 0.04) (P < 0.05) were noted. In addition, injection of pUDKH could alleviate the severity of degeneration of muscular tissue due to ischemia. CONCLUSION: Human HGF gene therapy is effective in treating rat acute limb ischemia with the lowest effective dose of 100 microg. PMID- 14521734 TI - [The transduction of neurotrophin-3/GDNF attenuates the SGNC damage in deaf mice]. AB - OBJECTIVE: To investigate the attenuation of cisplatin-induced toxicity in the spiral ganglion neuron (SGNC) by the localized expression of the neurotrophin-3 (NT-3) and glial cell line-derived nerutrophic factor (GDNF) via HSV-Amplicon. METHODS: We constructed the HSV-Amplicon vectors, which could express the NT-3 and GDNF under separate transcriptional control. Helper virus-free amplicon stocks were assessed in vitro for their capacity in the cultured inner ear cells. The ELISA was used to detect the production of NT-3 and GDNF. Three groups of mice were injected with cisplatin (8 mg/kg), followed by instillation of HSVnt 3myc/gdnf, HSVnt-3myc/lac or HSVlac. The numbers of SGNC were analyzed 4 weeks later. RESULTS: The HSVnt-3myc/gdnf transduction resulted in production of NT-3 up to 11.44 micro g/ml and GDNF up to 1.79 ng/ml in cultured medium over 48 h. There were significant differences among three groups (P < 0.01). Cochleae transduced with HSVnt-3myc/gdnf and HSVnt-3myc/lac harbored from the mice had greater number of surviving SGNC. The ratio of SGNC survival (%) in HSVnt 3myc/gdnf, HSVnt-3myc/lac and HSVlac was 88%, 77% and 22%, respectively. CONCLUSION: The transduction of NT-3/GDNF by HSV-Amplicon greatly attenuated SGNC the cisplatin-induced ototoxicity in mice. PMID- 14521736 TI - [Isopotential map of somatosensory evoked potential and motor revoked potential in spinal cord: an experimental study]. AB - OBJECTIVE: To investigate the distribution of conduction pathways of somtosensory evoked potential (SEP) and motor evoked potential (MEP) in the spinal cord. METHODS: Twenty-five Wistar rats underwent operation to expose the left sciatic nerve and sphenotresia. The sciatic nerve and sensory-motor cortex of 10 rats were stimulated so as to induce MEP and SEP. Different stimulating parameters were used to record their influence on MEP and SEP. Then the microelectrode was removed to a new place 1 cm from the original place and the maximum MEP and maximum SEP were recorded so as to calculate the conduction speeds of MEP and SEP. A microelectrode was put to the ventral side of spinal cord of 15 rats so as to record the isopotential map of MEP and SEP in the spinal cord. RESULTS: The Ni P1 of SEP increased when the microelectrode was moved from the anterior side to the posterior side and the change was more prominent in the anterior funiculus area. The N2-P2 decreased when the microelectrode was moved from anterior side to posterior side, and the change was more prominent in the posterior funiculus area. CONCLUSION: SEP is mainly conducted in the posterior tracts and MEP in the anterior tracts. MEP may originate from extra-pyramid system. PMID- 14521737 TI - [The intervention of reduced glutathione in neutrophil activation in the process of extracorporeal circulation]. AB - OBJECTIVE: To study the intervention of reduced glutathione for neutrophil activation in the process of extracorporeal circulation. METHOD: 20 congenital heart disease children aged from 6 to 12 years were selected randomly, divided into control group and interventional group, patients of interventional group were added reduced glutathione (30 mg/kg) in priming fluid, then phlebotomized on the time of 24 hours before operation, 20 minutes after CPB, rewarming, the end of CPB, 4 hours after operation, 24 hours after operation. After separated neutrophil, we distilled nuclear protein and mensurated the activation of nuclear factor-kappaB by EMSA, assayed the expression of ICAM-1. by immunohistochemistry. RESULTS: Neutrophil nuclear factor-kappaB has been activated on the time of 20 minutes after CPB, arrives at the top on the time of rewarming, and resumes 24 hours after operation; ICAM-1 expresses on the time of rewarming, arrives at the top 4 hours after operation in control group. The data of interventional group is lower than the control group. CONCLUSION: Neutrophil nuclear factor-kappaB is activated and ICAM-1 is expressed in the process of extracorporeal circulation and play an important role in the inflammatory response happening in extracorporeal circulation, which could be relieved by reduced glutathione. PMID- 14521738 TI - [Cardioprotective effects of diltiazem infusion in the perioperative period in patients undergoing coronary artery bypass grafting with extracorporeal circulation]. AB - OBJECTIVE: To evaluate the cardioprotective effects of intravenous diltiazem infusion perioperatively in patients undergoing coronary artery bypass grafting (CABG) under extracorporeal circulation. METHODS: Forty patients undergoing CABG were divided into 2 groups of 20 patients according to the operation date: experimental group in which infusion of diltiazem (0.1 mg/kg/h) was given starting from the onset of extracorporeal circulation until 24 h after the aortic cross-clamping, and blank control group. The relative hemodynamic parameters were recorded for 48 hours. RESULTS: (1) 4 approximately 24 hours after the aortic cross-clamping the heart rates of the experimental group was 98 +/- 18 approximately 96 +/- 13 times/min, significantly lower than those of the control group (111 +/- 16 approximately 105 +/- 14 times/min, P<0.05); the values of rate pressure product (RPP) of the experimental group were 13.0 +/- 3.7 approximately 11.7 +/- 3.0 mm Hg/min/10(3), significantly lower than those of the control group (14.1 +/- 1.9 approximately 5.8 +/- 3.6 approximately mm Hg/min/10(3), P<0.05) (2) The postoperative peak value of creatine kinase (CK) was not significantly different between these 2 groups. The postoperative peak value of creatine kinase MB (CK-MB) in the experimental group was 83 +/- 33 IU/L, significantly lower than that in the control group (105 +/- 20 IU/L, P<0.05) and postoperative peak value of cardiac troponin-T (cTNT) was 0.38 +/- 0.21 IU/L, significantly lower than that in the control group (0.83 +/- 0.20 IU/L, P<0.05). The values of CK, CK-MB, and cardiac troponin TC (cTNT) of the patients with coronary vasospasm in the experimental group were all significantly lower than those of the patients with coronary vasospasm in the control group. (3) The average number of arrhythmic events per hour was lower in diltiazem group, and the average number of ventricular premature contraction per hour was significantly lower during the period of ST segment depression in particular. CONCLUSION: Perioperative intravenous diltiazem infusion provides significant cardioprotective effect in patients undergoing CABG. PMID- 14521739 TI - [Myocardial protective effect of ulinastatin against ischemia/reperfusion injury during open heart surgery with cardiopulmonary bypass]. AB - OBJECTIVE: To investigate the protective effect of ulinastatin on myocardium against ischemia-reperfusion injury in open heart surgery with cardiopulmonary bypass (CPB). METHODS: Twenty ASA I-II patients undergoing atrioseptopexy or surgical repair of ventricular septal defect under CPB were randomly divided into two groups of 10 patients. The patients in the ulinastatin group (U), 5 males and 5 females, aged 6.7 +/- 2.6, received ulinastatin 12,000 unit/kg, half of the dose being given intravenously 10 min before aorta cannulation and another half being added into the priming fluid. The patients in the control group (C), 6 males and 4 females, aged 5.9 +/- 2.7, received the same volume of normal saline instead of ulinastatin. Arterial blood samples were taken before CPB (T1), at release of the aortic cross-clamp (T2), 30 min after aortic release (T3), 4 h and 24 h after discontinuation of CPB (T4, T5) for determination of plasma levels of cardiac troponin I (cTnI), creatine phosphokinase (CK) and creatine phosphokinase isoenzyme (CK-MB). RESULTS: The CPB time, aortic cross-clamping time and duration of operation were comparable between these 2 groups. The plasma cTnI level and CK and CK-MB activities were all within normal range before CPB in both groups. In group C the plasma level of cTnI started to increase at T2, peaked at T4 and started to decrease at T5. In group U the plasma levels of cTnI at T3 and T4 were significantly higher than the baseline value (both P<0.01) and returned to the baseline value at T5. The plasma cTnI levels at T(3-5) were significantly lower in group U than in group C (all P<0.01). The plasma CK and CK-MB activities increased significantly at T(2-5) in both groups (all P<0.01). There was no significant difference in plasma CK and CK-MB activity at T(2-4) between the two groups, but at T5 their activities were significantly lower in group U than in group C (P<0.05). The rate of spontaneous recovery of heart beat without defibrillation was higher in group U (8/10) than in group C (4/10) (P<0.05). The drainage volume during the 24 hours after operation was greater in group C than in group U (P<0.05). CONCLUSION: Ulinastatin effectively protects myocardium from ischemia-reperfusion injury during open heart surgery with CPB. PMID- 14521740 TI - [A preliminary study of synchronous evoked potential in newborn-infant auditory and visual function test]. AB - OBJECTIVE: The purpose of this study is to bring out the consultable laboratory values of synchronous evoked potential in normal neonates and infants, investigate the characteristic of the electric response waveform, and explore its feasibility on applying to newborn auditory and visual function test as an electric physiology detection method. METHODS: It was applied to go on measuring 19 cases of mature normal neonate (< or =4 days) and 11 cases of infants (< or =5 months), also comparing with the study of ABRs, FVEPs independently. RESULTS: Get the electric response wave form results of synchronous electric evoked potential in both groups of newborn-infants, through comparing with the result of independent ABRs and FVEPs, the monitoring result demonstrated that there were an extreme significant changes of the corresponding parameters (wave I, III, V latency) (P<0.01), and there were no significant changes of I-III, III-V interwave latency (P>0.05); it also shows that ABRs impact on FVEPs in N2 wave latency of newborn group and P1 wave latency of infant group (P<0.01). CONCLUSION: Auditory and vision pathway may be had communicated information by the neuron in the center. The technology of the auditory and visual synchronous evoked potential brings out the whole new electric physiology detection tools, it will definitely offer powerful technical support in step for newborn-infant auditory and visual function test. PMID- 14521741 TI - [A study on the risk indicators of newborn hearing loss]. AB - OBJECTIVE: To explore the risk indicators of newborn hearing loss. METHODS: Statistic description was used to analyze the basic characteristics of 41 hearing loss infants screened from 8,262 newborns; one way analysis was used by 1:2 matched case control study to analyze the risk indicators of newborns with hearing loss; chi-square test and multivariate condition Logistic stepwise regression mode were also used to find risk the indicators. RESULTS: By one way analysis, there were 3 high risk indicators associated with newborn hearing loss: family history of hearing loss, craniofacial anomalies, and NICU care history. By analysis of multivariate condition Logistic stepwise regression mode, family history of hearing loss (OR=16.945, 95% CI 1.21-237.369) and craniofacial anomalies (OR=6.703, 95% CI 0.61-73.85) were 2 independent risk factors of newborn hearing loss. CONCLUSION: Appropriate intervention measure should be done to reduce the high risk indicators that cause newborn hearing loss. PMID- 14521742 TI - [Complications of stent-assistant angioplasty of symptomatic intracranial artery stenosis]. AB - OBJECTIVE: To discuss the safety of intracranial stenting for refractory symptomatic intracranial artery stenosis. METHODS: Forty-eight patients with symptomatic intracranial artery stenosis were treated by transluminal stent assistant angioplasty. Of them, 40 cases were selected because they had recurrent TIAs or mild stroke despite of antiplatelet or anticoagulation therapy; 8 cases with high-grade stenosis after acute cerebral artery theromblysis. Lesions involved MCA (17/48); Basilar artery (8/48); intracranial vertebral artery (18/18); and distal ICA (5/18). RESULTS: For 46 of 48 cases the flexible coronary stent were successfully deployed. The average stenosis reduced from 83% to 5%, short-term follow-up showed good clinical improvement. Complications include vessel rupture (1/18); acute thrombosis within stent (1/48); perforation of cortical artery (1/18) and perforate vessel occlusion (1/18). CONCLUSION: Stent assistant angioplasty are effective for treatment of symptomatic intracranial stenosis, the higher rate of complications may be because of the limited experiences of this technique. it need further practise and long term follow-up study. PMID- 14521743 TI - [A randomized comparative study of naphtoquine, mefloquine and artsunate in the treatment of falciparum malaria]. AB - OBJECTIVE: To evaluate the efficacy and safety of naphtoquine, compared with mefloquine and artesunate in the treatment of falciparum malaria. METHOD: Ninety patients with falciparum malaria were randomly allocated to 3 groups, including naphtoquine, mefloquine and artesunate group. In the naphtoquine group, thirty patients were prescribed single daily dosage of 1,000 mg for one day. In the mefloquine group, equal patients were treated with single dosage of 750 mg. Another thirty patients in the artesunate group were given total dosage of 600 mg for five days and doubling dosage on the first day. RESULT: In all three groups, symptoms were well controlled. The average fever-subsidence time in naphtoquine group was 30 h +/- 16 h and approximate that in mefloquine group (24 h +/- 15 h, P>0.05), but was longer than that in naphtoquine group (18 h +/- 9 h, P<0.01). The average parasite-clearance time in naphtoquine group (98 h +/- 28 h) is longer than that in mefloquine group (57 h +/- 20 h, P<0.01) and that in artesunate group (43 h +/- 17 h, P<0.01). At the end of 28-day clinical trail, the curative ratio in naphtoquine group was the highest (96.7%), and was significantly higher than that in mefloquine group (76.7%, P<0.05) and artesunate group (73.3%, P<0.05). Slight nausea and vomiting were observed in few patient in three groups. CONCLUSION: Although the average fever-subsidence time and the parasite-clearance time of naphtoquine at single 24-hour dosage of 1,000 mg were longer than those of mefloquine and artesunate, the 28-day curative ratio of naphtoquine was higher than that of mefloquine and artesunate. So we recommend that the combination of artemisinin, which is a rapid action antimalarial, and naphtoquine or mefloquine, which are longterm action antimalarial, would contribute to promoting efficacy, shorting the period of treatment and delaying occurrence of drug resistance. PMID- 14521744 TI - [Influence of beta-endorphin on function of immune system of patients with cerebral hemorrhage]. AB - OBJECTIVE: To study the influence of beta-endorphin (beta end) on the function of immune system of patients with cerebral hemorrhage at different stages. METHODS: Radioimmunal analysis was applied to detect the serum beta-endorphin concentration in the peripheral blood of 28 patients with cerebral hemorrhage, aged 65.5 +/- 13, 28 age-matched patients with cerebral thrombosis, and 28 sex and age-matched normal controls. Mononuclear cells from peripheral blood of these 3 kinds of subjects were cultured and then beta end 10(-8) g/L, beta end 10(-11) g/L, beta end 10(-14) g/L, or beta end 10(-11) g/L + naloxone 10(-5) g/L were added into the media respectively and the MNCs were cultured for more 24 hours (beta end 10(-8) g/L group, beta end 10(-11) g/L group, beta end 10(-14) g/L group, and beta end 10(-11) g/L + Nal group). Another MNCs were cultured without addition of beta end (beta end 0 g/L group). Then the MNCs were collected. RT-PCR was used to detect the expressions of interleukin (IL)-1beta, IL-2, IL-8 and iNOS mRNA in the MNCs. RESULTS: The serum beta-end level of the patients with cerebral hemorrhage at the acute stage was 129 +/- 82 ng/L, significantly lower than that of the normal controls (321 +/- 62 ng/L, P<0.01) and that of the patients with cerebral thrombosis (264 +/- 163 ng/L, P<0.05), but not significantly different from that of the patients with cerebral hemorrhage in the convalescent stage (160 +/- 72 ng/L, P>0.05). The expression of IL-1beta and the expression of IL-2 of the patients with cerebral hemorrhage at the acute stage were significantly lower than those of the patients with cerebral thrombosis and the controls (all P<0.01). The expression of IL-1beta of the patients with cerebral hemorrhage at the convalescent stage were higher than that in the acute stage, however, the difference was not significant (P>0.05). The expression of IL-2 of the patients with cerebral hemorrhage at the convalescent stage was higher than that at the acute stage (P<0.01). The expression of IL-8 and the expression of iNOS of the patients with cerebral hemorrhage at the acute stage were significantly higher than those of the patients of cerebral thrombosis and the controls (both P<0.01). The expression of IL-8 and the expression of iNOS of the patients with cerebral hemorrhage at the convalescent stage were significantly lower than those in the acute stage (both P<0.01). The expressions of IL-1beta, IL-2, IL-8, and iNOS mRNA in the peripheral blood MNCs in vitro in the beta end 10(-8) g/L group and beta end 10(-11) g/L group were significantly higher than those of the beta end 0 g/L group, beta end 10(-11) g/L group, and beta-end 10(-11) g/L + Nal 10(-5)g/L group. The expressions of IL-1beta, IL-2, IL-8, and iNOS mRNA in the peripheral blood MNCs in the beta-end 10(-11) g/L +Nal 10(-5) g/L group were higher than those of the beta end 0 g/L group, however, not significantly. CONCLUSIONS: The endogenous beta-endorphin level of cerebral hemorrhage patients is low. The immune system function is up-regulated at the acute stage and then down regulated. Thereafter the immune system function is invariably low. Exogenous beta-endorphin enhances the IL-1 beta, IL-2, IL-8 and iNOS mRNA expression of peripheral blood MNCS. beta-endorphin receptor antagonist naloxone blocks the positive immunoregulation by beta-endorphin. PMID- 14521745 TI - [Prevalence of low vision and blindness in defined populations in rural and urban areas in Beijing]. AB - OBJECTIVE: To investigate the prevalence and causes of low vision and blindness in Beijing residents aged 40 and over. METHODS: 4,451 residents aged 40 and over in 3 rural communities and 5 urban communities in Beijing underwent eye examination, including examination of distant and near visual acuity (VA), best corrected distant and near VA, pinhole VA, and visual field, slit lamp biomicroscopy, and dilated ocular examination in the form of in-home survey by defined population-based sampling. The medical history was surveyed too. The data were analyzed based on the criteria of the World Health Organization. RESULTS: The general prevalence rates of low vision and blindness were 0.99% (95% CI: 0.70 1.28) and 0.39% (95% CI: 0.21-0.57) respectively. The prevalence rate of low vision in females was 1.45%, 2.23 times that of males (0.65%) (OR: 1.97, 95% CI: 1.00-3.95). The prevalence rate of low vision of rural residents was 1.76%, 2.89 times that of urban residents (0.61%) (OR: 2.93, 95% CI: 1.43-6.11). The prevalence rate of blindness in females was 0.64% and 0.37% in males. The prevalence rate of blindness of rural residents was 1.06%, 2.04 times that of the urban residents (0.52%) (OR: 3.77, 95% CI: 1.41-10.62). The 3 major causes of blindness were cataract (37.50%), glaucoma (29.20%), and high myopic macular degeneration (8.30%). The prevalence of blindness increased with age. CONCLUSION: The prevalence rates of low vision and blindness are higher in the rural areas. Cataract, glaucoma, and high myopic macular degeneration are the major causes of blindness. The prevalence of low vision and blindness are influenced by age, sex; area, health care level, educational level, and environmental factors. PMID- 14521746 TI - [The differential diagnosis between uterine leiomyosarcoma and the special subtypes of leiomyoma]. AB - OBJECTIVE: To explore the more sensitive and specific immunohistochemical markers in differential diagnosis of uterine leiomyosarcoma (ULMS) and the specific subtypes of leiomyoma. METHODS: The routine SP immunohistochemical technique was used to examine the expression of desmin, alpha smooth muscle actin (SMA), calponin h1, h-caldesmon, estrogen receptor (ER), progesterone receptor (PR), proliferating cell nuclear antigen (PCNA), mast cells, and CD44v3 in 82 patients with smooth muscle tumors of the uterus, including 17 patients with leiomyosarcoma (LMS), 25 with cellular leiomyoma (CL), 15 with bizarre leiomyoma (BL) and 25 with ordinary leiomyoma (OL). RESULTS: The expression rates of desmin, SMA, h-caldesmon and CD44v3 in uterine LMS were all lower than those in CL and BL (all P<0.05), but these markers had limited significance in differentiation of ULMS from CL or BL. Compared to those in CL or BL, the expression rates of ER and PR and the number of mast cells were significantly lower in LMS (all P<0.01), while the expression of PCNA in LMS was higher (P<0.01). CONCLUSION: The increase of PCNA and decrease of ER and PR expression help differentiate ULMS from CL and BL. The number of mast cells was helpful in differential diagnosis of ULMS with high sensitivity and specificity. PMID- 14521747 TI - [Effect of motilin on gastric smooth contraction induced by interstitial cells of Cajal]. AB - OBJECTIVE: To investigate the role of interstitial cells of Cajal (ICC) in contraction of gastric smooth muscle and the effect of motilin on this role. METHODS: Two kinds of smooth muscle segments with ICC were isolated from the pacemaker area at the upper 1/3 of stomach body and from the gastric antrum of 90 rats and immersed into Krebs solution and linked with tonotransducer to record their contraction. Both kinds of smooth muscle segments were divided into 5 groups equally: group A (control group), group B (methylene blue and intense illumination were used to destroy the ICC therein), group C (motilin was added into the Krebs solution), group D (both motilin and anti-motilin serum were added), and group E (motilin and atropin or hexahydroxy-quaternary ammonium were added). RESULTS: stable contractile activity was recorded in these 2 kinds of smooth muscle segments with higher frequency and amplitude of contraction in those from the gastric antrum than those from the stomach body. The frequency and amplitude of contraction in the smooth muscle segments with the ICC destroyed by methylene blue and intense illumination were decreased by 80% approximately 90%. Motilin increased the frequency and amplitude of contraction in the smooth muscle segments from the gastric antrum and stomach body dose-dependently. Anti-motilin serum and atropin, but not hexahydroxy-quaternary ammonium blocked the effect of motilin. The smooth muscle segments with destroyed ICC showed no significant differences in the frequency and amplitude of contraction in comparison with the smooth muscle segments with destroyed ICC plus motilin. CONCLUSION: The pacemaker cells of the contractile activity of gastric smooth muscle cells are ICC, Motilin regulates the gastric smooth muscle contraction initiated by ICC. PMID- 14521748 TI - [The expression of connective tissue growth factor in renal cortex of 5/6 nephrectomized rats and its modulation by fluvastatin]. AB - OBJECTIVE: To investigate the connective tissue growth factor (CTGF) mRNA expression in the renal cortex of 5/6 nephrectomized rats and its modulation by fluvastatin, a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor. METHODS: Twenty-four rats underwent operation 2 times: during the first operation 2/3 of the left kidney was resected, and the right kidney was resected completely one week after. The 24 5/6 nephrectomized rats were randomly divided into 2 groups: untreated 5/6 nephrectomized group (model group, n=12) and fluvastatin treated 5/6 nephrectomized group (treatment group, fluvastatin was orally administered 7 mg.kg(-1).d(-1) for 13 weeks, n=12), and 6 sham-operated rats served as control (sham operation group). In the weeks 2, 4, 8, and 13 after the second operation metabolic cage was used to collect the 24-hour urine 2 times. Urine protein was examined by biuret reaction so as to calculate urinary protein excretion. By the end of experiment blood was collected to examine the serum cholesterol, triglyceride, urea nitrogen, and creatinine contents. The rats were killed and their kidneys taken out. The CTGF mRNA expression in the renal cortex was detected by RT-PCR. Immunohistochemistry was used to examine the expression of transforming growth factor-beta1 (TGF-beta1), type IV collagen and fibronectin in the glomeruli. Renal pathological changes and glomerular sclerosis index (GSI) were evaluated as well. RESULTS: At the end of the experiment, the mean urinary protein excretion in the model group was 305.4 mg/24 h, significantly higher than that in the treatment group (230.9 mg/24 h, P<0.01) and the sham operation group (5.6 mg/24 h, P<0.01) The serum urea nitrogen of the model group was (24.5 +/- 4.9) mmol/L, significantly higher than that of the treatment group [(15.8 +/- 3.9) mmol/L, P<0.05] and that of the sham-operated group (7.4 +/- 0.3 mmol/L, P<0.01). The serum creatinine (P<0.05) of the model group was 88 micromol/L +/- 14 micromol/L, significantly higher than that of the treatment group [(58 +/- 5) micromol/L, P<0.05)] and that of the sham-operated group [(54 +/- 5) micromol/L, P<0.05]. The creatinine clearance rate of the model group was (1.7 +/- 0.7) ml.min(-1).kg(-1), significantly lower than that of the treated group [(3.2 +/- 1.1) ml.min(-1).kg(-1), P<0.05] and that of the sham-operated group [(3.9 +/- 1.5) ml.min(-1).kg(-1), P<0.05]. The glomerular sclerosis index (GSI) in the model group was 41.8 +/- 11.5, significantly higher than that in the sham operation group (2.2 +/- 1.3, P<0.01) and the treatment group (23.4 +/- 6.1, P<0.05). The mean optical density of CTGF mRNA expression in the renal cortex of the model group was a 3 times that of the sham operation group, and the mean optical density of CTGF mRNA expression in the renal cortex of the treatment group was lower by 55.4% compared with that of the model group (P<0.01). The glomerular expressions of TGF-beta1, type IV collagen and fibronectin were significantly up-regulated in the model group in comparison with those in the sham operation group (all P<0.01). The glomerular protein expressions of TGF beta1, type IV collagen and fibronectin were significantly weaker in the fluvastatin treatment group as compared with the model group (all P<0.01). CONCLUSION: CTGF mRNA expression is markedly upregulated in the renal cortex of 5/6 nephrectomized rats. Fluvastatin suppresses the increased CTGF mRNA expression in renal cortex and ameliorates the glomerular extracellular matrix accumulation. PMID- 14521749 TI - [Sensory neuropeptide SP modulating expression of EGF, FGF-2 and their receptors in fibroblasts of granulation in vivo and in vitro]. AB - OBJECTIVE: To study the modulating effect of sensory neuropeptide substance P (SP) on gene expression of epidermal growth factor (EGF), fibroblast growth factor-2 (FGF-2) and EGF receptor (EGFR), FGF receptor-1 (FGFR-1) in granulation fibroblast. METHODS: The study consisted of two parts, in vivo and in vitro. In vivo, fifty wistar rats were randomly arranged into capsaicin group and control group. The rats in capsaicin group were subcutaneously injected with neurotoxin dose of capsaicin (50 mg/kg) on the back of the rats to chemically destroy sensory nerves to prevent the release of SP. After one week, full-thickness skin wounds were created on the rats' back. SP content and EGF, EGFR, FGF-2, FGFR-1 gene expression in the granulation tissue were observed using immunohistochemistry and in situ hybridization combined with image analysis on 3rd, 6th, 9th 12th after skin injury, respectively. The experimental procedure in control group were similar with that in capsaicin group except injection with capsaicin. In vitro, SP (10(-9)-10(-5) mol/L) was added into culture medium for cultured fibroblasts from granulation and mRNA expressions of EGF, FGF-2 and their receptors were assayed with RT-PCR. RESULTS: In the present study, immunoreactive stain for SP in granulation tissue were correlated with gene expression of EGF, FGF-2 and their receptors using in situ hybridization in vivo. Once sensory nerves were destroyed and SP release was inhibited in capsaicin group, the gene expression of above growth factors and their receptors were attenuated. SP concentrations in culture medium for upregulation of mRNA expressions of EGF and EGFR in cultured fibroblasts were from 10(-8) mol/L to 10( 6) mol/L and from 10(-6) mol/L to 10(-5) mol/L, respectively. SP concentration was from 10(-9) mol/L to 10(-5) mol/L for upregulation of FGF-2 mRNA expression in cultured fibroblasts and from 10(-6) mol/L to 10(-5) mol/L for upregulation of FGFR-1. CONCLUSION: Sensory neuropeptide SP released from sensory nerve participates in upregulation of gene expressions of EGF, EGF-2 and their receptors in granulation fibroblasts during wound healing. PMID- 14521750 TI - [Transplantation of cord blood endothelial progenitor cells ameliorates limb ischemia]. AB - OBJECTIVE: To investigate the feasibility of transplanting cord blood CD133+ cells derived endothelial progenitor cells (EPC) in therapeutic vasculogenesis. METHODS: CD133+ cells from the cord blood of 52 neonates were cultured in fibronectin-coated flask in M199 medium supplemented with 10% fetal bovine serum, 50 ng/ml vascular endothelial growth factor (VEGF), 20 ng/ml interleukin-3 (IL-3) and 50 ng/ml stem cell factor (SCF). The cell markers of spindle-shaped adherent cells were determined with flow cytometry. The left femoral artery, great saphenous artery, iliac circumflex artery, and vein, and muscular branch of 22 Balb/c nude mice were cut to cause limb ischemia. One day after the unilateral ischemic limb surgery half million adherent cells were transplanted into 12 nude mice via tail vein (EPC group) and M199 was injected into the tail veins of 10 nude mice (M199 group). Fluorescence Dil, laser Doppler perfusion imaging (LDPI) and immunohistochemistry were Laser Doppler perfusion imaging (LDPI) was used to trace the transplanted cells and monitor the blood perfusion and capillary density of ischemic limbs. The ratio between the blood perfusion of the operated limb and of the non-operated opposite limb was recorded. Two to four mice in each group were killed 4, 7, 14, and 21 days after the operation and the gastrocnemius muscles of bilateral hind limbs were taken to count the number of capillaries. The VEGF mRNA levels of the ischemic and nonischemic limbs were examined with semi-quantitative RT-PCR. Seven days after the operation, fluorescein isothiocyanate (FITC)-binded ulex europaeus agglutinin-1 (UEA-1) was injected via tail vein to 3 EPC group mice. Thirty minutes later, the mice were killed. The heart, lung, liver, spleen and limb muscles were taken and examined with fluorescence microscopy. EPC were added into the upper chamber of Coster Transwell and chemotactic fluids of M199 with or without VEGE were added into the lower chamber. Four hours later the number of EPC in the lower chamber was counted so as to examine the chemotactic effect of VEGE. RESULTS: Numerous cell clusters, spindle-shaped adherent cells and cord-like structures, developed from the culture of cord blood CD133+ cells. These adherent cells expressed vascular endothelial growth factor receptor 2 (VEGFR-2), VE-cadherin, CD31, von Willebrand factor (vWF) and combined with ulex europaeus agglutinin-1 (UEA-1). Transplanted EPC survived and were incorporated into the capillary networks in the ischemic limbs of nude mice. The ratio between the blood perfusion of the ischemic limb and non-ischemic limbs was 19.1% +/- 3.1%. Two weeks after the transplantation, the ratio between the blood perfusion of the ischemic limb and non-ischemic limbs of the EPC group was 77.3% +/- 5.6%, significantly higher than that of the M199 group (40.6% +/- 3.4%, P<0.001). CD31 histochemical staining showed that the density of capillaries in the gastrocnemius muscles of ischemic hind limb was significantly higher 7, 14, and 21 days after operation in the EPC group than in the M199 group (P<0.05) RT-PCR showed obvious VEGF bands in the ischemic hind limb muscles, but not in the non-ischemic muscles. The number of EPC immigrating into the lower chamber of the Coster Transwell was 817 +/- 32.5, significantly higher than that of the control group (473.5 +/- 61.5, P<0.05). CONCLUSION: Cord blood CD133+ cells derived EPC is a robust cell source for therapeutic neovascularization. Upregulated expression of VEGF may account for the homing of transplanted EPC to ischemic tissue. PMID- 14521751 TI - [The effect of specific virus delivering of Langerhans cell for the M-tropic HIV infection]. AB - OBJECTIVE: To observe the cellular mechanism of via skin infection of Microphage tropic (M-tropic) HIV and further differentiate the infection of M-tropic and T tropic HIV. METHODS: DNA clone of AD8 and NL43 was respectively transfected to skin by Gene Gun, The expression of virus in the epidermal cells was observed by flow-cytometry and fluoroscope. Emigrating cells were Mac-sorted into CD80+ and CD80-. The infection of CD4+ T lymphocytes by HIV from emigrating cells were further assessed by PCR and RT assay. RESULTS: Through direct transfection of skin by Gene Gun, emigrating cells from skin showed transfection of AD8 and NL43 and the gag gene can be amplified from the cells. Only AD8 transfected emigrating cells can further infect CD4+ T lymphocyte. Gag can be amplified and reverse transcriptase can be expressed from these CD4+ T cells; NL43 transfected emigrating cells can not infect CD4+ T lymphocyte. No gag can be amplified and no reverse transcriptase can be expressed. AD8 transfected and 1/2 log diluted CD80+ cells can infect CD4+ T lymphocyte. Even after 4 dilutions, there are still a lot of CD4+ T cells, gag can be amplified and a large amount of reverse transcriptase was expressed that is double or triple of controls. AD8 infected CD80-cells co cultured with CD4+ T cells showed gag amplification only in first dilution and reverse transcriptase is also expressed. Gag can not be amplified by further dilution. NL43 infected CD80+ or CD80- emigrating cells co-cultured with CD4+ T cells did not show gag amplification and did not express reverse transcriptase. CONCLUSION: Direct AD8 transfected CD80+ emigrating epidermal cells can specifically infect CD4+ T lymphocyte. This specificity not only depends on affinitive combination of HIV receptor on cell surface, but also on the specific virus delivering of AD8 transfected CD80+ cells that infect the CD4+ T cells. That means even without the receptor, CD80+ cells only deliver transfected AD8 but not NL43. So, it can be said that the specificity of the virus receptor is not the only clue of HIV infection. PMID- 14521752 TI - [An investigation of the relationship between Lepr gene Gln223Arg polymorphism and obstructive sleep apnea hypopnea syndrome]. AB - OBJECTIVE: To investigate the relationship between leptin receptor (Lepr) gene Gln223Arg polymorphism and obstructive sleep apnea hypopnea syndrome (OSAHS). METHODS: The genotypes of Gln223Arg polymorphism in Lepr gene were determined by polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) assay in 181 unrelated subjects of North region "Han" population (including 78 non-OSAHS subjects and 103 OSAHS subjects). RESULTS: The OSAHS patients with GG genotype had significantly larger neck circumference (NC) than did OSAHS patients with (GA + AA) genotype (P = 0.031); The study revealed no difference in genotype of Gln223Arg polymorphism between OSAHS group and non-OSAHS group (P = 0.676); There was also no significant difference in body mass index (BMI), waist hip rate (WHR), blood pressure (Bp), leptin (Lep), true insulin (TI), fast blood glucose (FBG), triglyceral (TG), cholesterol (Chol) and polysomnography (PSG) between GG and (GA + AA) genotype of OSAHS patients (P > 0.05). CONCLUSIONS: Lepr gene Gln223Arg polymorphism may be involved in the regulation of the neck regional fat distribution in OSAHS patients. It is unlikely that Lepr gene Gln223Arg polymorphism may contribute to the pathogenesis of OSAHS. PMID- 14521753 TI - [Respiratory control during sleep and the development of nocturnal hypoxia in patients with obstructive sleep apnea hypopnea syndrome]. AB - OBJECTIVE: It is assumed that the reduced ventilatory control during sleep may be related to the nocturnal hypoxia in patients with obstructive sleep apnea hypopnea syndrome (OSAHS) and daytime CO(2) retention. METHODS: Oxygen desaturation index (ODI(4)), sleep time spent when SaO(2) was below 90% (SIT(90)), the lowest SaO(2) (LSaO(2)) and the mean SaO(2) (MSaO(2)) during sleep were measured in 24 OSAHS patients with daytime PaCO(2) > or = 45 mm Hg and 39 with PaCO(2) < 45 mm Hg. Hypoxic (DeltaP(0.1)/DeltaSaO(2)) and hypercapnic (DeltaP(0.1)/DeltaPaCO(2)) responsiveness were measured in 11 eucapnic and 4 hypercapnic patients during wakefulness, NREM and REM sleep stages. RESULTS: Compared with the eucapnic patients, all the hypercapnic patients had more severe hypoxia during sleep (P < 0.05). Although both groups had similar chemical responsiveness during wakefulness, the hypercapnic patients had lower DeltaP(0.1)/DeltaSaO(2) and DeltaP(0.1)/DeltaPaCO(2) during both REM and NREM sleep (P < 0.05). CONCLUSION: The depressed chemical responsiveness during sleep plays an important role in the development of nocturnal hypoxia in OSAHS patients with daytime CO(2) retention. PMID- 14521754 TI - [Relationships between the diameters of Bacille Calmette-Guerin scars and asthma, atopy in urban and rural Beijing children]. AB - OBJECTIVE: To compare the diameters of Bacille Calmette-Guerin (BCG) scars between asthmatic (allergic) students and normal students, and to compare the diameters of BCG scars in children in rural Beijing area with those in urban Beijing area, and to compare the prevalence of asthmatic (allergic) symptoms and atopy between rural and urban Beijing students. METHODS: Sampled by cluster random Method, 819 urban school children and 1 044 rural school children aged 13 14 years were recruited for the Beijing arm of the phase III international study of asthma and allergies in childhood. The children answered the core questionnaires, and underwent skin prick test for 13 allergens, and were checked for BCG scars on their arms. RESULTS: (1) In both rural and urban Beijing students, the mean diameters of BCG scars in those with asthma (allergy) (asthma ever, wheeze ever, current wheeze within 12 months, hay fever ever, repeated rash ever, eczema ever, allergic rhinitis, atopy, high-sensitivity or were diagnosed asthma) were (7.0 +/- 2.1) mm-(8.8 +/- 2.8) mm, which were not significantly different from those of (7.2 +/- 2.7) mm-(8.2 +/- 2.2) mm in normal students (P > 0.05). (2) Compared to the urban students, the mean diameter of BCG scars in the rural students was (8.2 +/- 2.3) mm, significantly larger than that of (7.3 +/- 2.9) mm in the urban students (P < 0.01). The prevalence of asthma (allergy) (asthma ever, wheeze ever, current wheeze within 12 months, hay fever ever, repeated rash ever, eczema ever, allergic rhinitis, atopy, high-sensitivity or diagnosed asthma) were 0.6% - 11.4% in the rural Beijing students, significantly lower than that of 3.1% - 35.7% in urban Beijing schoolchildren (P < 0.01). CONCLUSIONS: The diameters of BCG scars were not significantly different between asthmatic (allergic) students and normal students. The diameters of BCG scars in the rural students were significantly larger than those in the urban students. The prevalence of asthma and allergy in the rural children were significantly lower than that in the urban students. The results suggest that there is difference in immunity between rural and urban children from early life, which may be caused by the different living environments. PMID- 14521755 TI - [Inhibition of antisense-endothelin converting enzyme RNA on interleukin-5 released from dust mite-challenged peripheral blood mononuclear cells in patients with allergic asthma]. AB - OBJECTIVE: To investigate the effect of antisense endothelin converting enzyme (ECE) RNA on levels of cytokines released from CD(4)(+) lymphocytes in patients with allergic diseases responsive to house dust mites. METHODS: Peripheral blood mononuclear cells (PBMCs) were separated from 21 patients who were sensitive to dust mites. PBMCs from those patients were divided into two groups. No stimulation group (A group) induded A(1) group (anti-ECE epithelial cells + PBMCs) and A(2) group (control cells + PBMCs). Stimulation group (B group) included B(1) group (anti-ECE epithelial cells + PBMCs + dust mites extract) and B(2) group (control cells + PBMCs + dust mites extract). House dust mite extract (20 microg/ml) was added to the culture of stimulation group as described above. After 72 hours, supernatants from both groups were collected and the levels of IL 5 and IFN-ggr; released into the supernatants were detected by enzyme-linked immunoabsorbent assay. RESULTS: IL-5 levels were increased significantly after treatment with dust mite in twelve of 21 cases. No significant differences of IL 5 were found between the groups of A(1)[(6.0 +/- 1.3) x 10(-9) g/L] and A(2) [(7.5 +/- 1.1) x 10(-9) g/L] before house dust mite stimulation in the 12 cases (P > 0.05), and no significant differences in IFN-ggr; were found between the groups of A(1) [(63 +/- 26) x 10(-9) g/L] and A(2) [(70 +/- 52) x 10(-9) g/L] before house dust mite stimulation (P > 0.05). IL-5 level was increased in both groups after stimulation but it was significantly lower in the B(1) group [(8.2 +/- 1.6) x 10(-9) g/L] than that in the B(2) [(12.0 +/- 1.8) x 10(-9) g/L] (P = 0.047). It seemed that increased IFN-ggr; level after stimulation was higher in B(2) [(153 +/- 71) x 10(-9) g/L] than that in the B(1) group (100 +/- 41) x 10( 9) g/L], but there was no statistic significance (P > 0.05). In addition, our results also showed that the release of IL-5 was significantly increased in those cases with asthma, or asthma plus allergic rhinitis after dust mites stimulation [(44 +/- 15)%] compared with that in those with urticaria [(7 +/- 4)%] (P = 0.047). CONCLUSIONS: Antisense-ECE downregulated the IL-5 secretion from Th2 lymphocytes in patients with allergic asthma after being challenged with dust mites. It is indicated that ET-1 is an important cytokine involved with allergic airway inflammation. Antisense-ECE RNA management in airways may be of value in treating allergic asthma. PMID- 14521756 TI - [Influence of hypoxia on peroxisome proliferator activated receptor alpha and its mechanisms]. AB - OBJECTIVE: To study the expressions of peroxisome proliferator activated receptor alpha (PPAR-alpha) and hypoxia-inducible factor-1alpha (HIF-1alpha) during different hypoxia states in lung cancer cell A549, and to investigate the mechanism of HIF-1alpha induced PPARalpha expression. METHODS: Firstly, lung cancer cells (A549) were divided into four groups: group A (control group); group B (hypoxia 24 h); group C (hypoxia 48 h); group D (hypoxia 72 h). The mRNA and protein of PPAR-alpha and HIF-1alpha were detected by RT-PCR and Western-blot on different groups. Secondly, the lung cancer cells (A549) were further divided into four groups: group E (control group); group F (antisense group); group G (sense group); group H (false-sense group). The influence of antisense oligonucleotide HIF-1alpha on PPAR-alpha was detected by RT-PCR and Western-blot. RESULTS: Comparing with group A, the expressions of the mRNA and protein of PPAR alpha and HIF-1alpha increased significantly in group B, and reached peak in group D. Conversely, the expressions of the mRNA and protein of PPAR-alpha were decreased after antisense oligonuclei of HIF-1alpha in group F, as compared with group E. CONCLUSIONS: The expressions of the mRNA and protein of PPAR-alpha and HIF-1alpha are increased by hypoxia, and PPAR-alpha may be regulated by HIF 1alpha. PMID- 14521757 TI - [Vascular endothelial growth factor inhibits dendritic cells from patients with non-small cell lung carcinoma]. AB - OBJECTIVE: To detect the effect of vascular endothelial growth factor (VEGF) on dendritic cells (DC) in patients with non-small cell lung cancer (NSCLC). METHODS: The measurement of DC in the peripheral blood was performed by a novel flow cytometric assay in 85 patients with NSCLC and 14 healthy volunteers. Enzyme linked immunosorbent assay (ELISA) was used to measure the concentration of VEGF(165) in the plasma. CD(14)(+) peripheral blood mononuclear cells (PBMC) were cultured to obtain DC in vitro with cytokines. VEGF(165) was added to evaluate its effect on DC differentiation and survival. The phenotypes and apoptosis of cultured cells were detected by flow cytometry. RESULTS: In comparison with healthy volunteers, the level of VEGF(165) was significantly increased (P < 0.05), while that of DC was significantly decreased (P < 0.01) in patients with NSCLC. No significant correlation was noted between the concentration of VEGF(165) and age, gender, differentiation and histological types in patients with NSCLC, neither was found in the level of DC (P > 0.05). The concentration of VEGF(165) was closely associated with TNM stage and distal metastasis (P < 0.05), while no correlation was found between the concentration of VEGF(165) and lymph node metastasis (P > 0.05). Significant correlations were noted between the level of DC in patients with NSCLC and TNM stage, lymph node metastasis and distal metastasis (P < 0.05). There was a negative correlation between the concentration of VEGF(165) and the level of DC (P < 0.05). Patients with abnormally elevated VEGF(165) showed significantly fewer DC. Cells cultured in vitro in the presence of VEGF(165) exhibited higher expression of CD(+)(14)(P = 0.000) and increased ratio of apoptic cells (P < 0.01), but decreased expression of CD(40), CD(86) and HLA-DR (P < 0.01), as compared to cells cultured without VEGF(165). CONCLUSIONS: The level of DC and the concentration of VEGF in the peripheral blood can reflect the malignancy of NSCLC. NSCLC can over-express VEGF to inhibit DC differentiation and maturation to evade host immune surveillance. PMID- 14521758 TI - [Investigation of drug-resistance to rifampin and rpoB gene sequence analysis of Mycobacterium abscessus]. AB - OBJECTIVE: To observe the resistance of Mycobacterium abscessus to rifampin and to investigate if there is any mutation of rpoB gene in strains with high minimal inhibitory concentration (MIC). METHODS: Mycobacterium abscessus was identified with both biochemical methods and PCR-RFLP. The MICs of rifampin to all the clinical strains and the type strain ATCC19977 were determined. DNA sequences were obtained from a 1272 bp fragment of the rpoB gene from either low or high rifampin MIC strains using PCR amplification. RESULTS: Only one strain had low MICs of 4 microg/ml. MICs of all the other 14 strains, as well as the type stain were above 128-256 microg/ml. Although there were some differences in nucleotide sequence of rpoB gene, all strains had the same amino acid sequence without any mutations. CONCLUSIONS: Mycobacterium abscessus showed a high resistance to rifampin. Mutations of rpoB gene do not seem to be the responsible mechanism. PMID- 14521759 TI - [A controlled clinical study on the efficacy of recombinant human interleukin-2 in the treatment of pulmonary tuberculosis]. AB - OBJECTIVE: To study and evaluate the efficacy and safety of recombinant human interleukin-2 (IL-2) in the treatment of pulmonary tuberculosis. METHODS: Two hundred and nine cases with re-treated Mycobacterium tuberculosis-positive pulmonary tuberculosis were randomly divided into a trial group (106 cases, treated with 3PaZ (TH)L(2)VE(AK) + IL-2/4PaL(2)V) and a control group (103 cases, treated with 3PaZ(TH)L(2)VE(AK)/4PaL(2)V). The efficacy of 203 cases was available for evaluation when the course was completed (trial group 103 cases, control group 100 cases). RESULTS: The sputum smear-negative conversion rates at the 1st and the 2nd month of therapy were 33.3% and 69.4% in the trial group, and 7.2% and 44.9% in the control group (P < 0.01). At the completion of the therapy, the X-ray resolution rates were 64.1% and 36.0% respectively for the trial and the control groups, the difference being significant (P < 0.001). There were significant differences in CD(4) T cells, the ratio of CD(4)/CD(8) and NK cells between the two groups (P < 0.01). The level of soluble interleukin-2 receptor (sIL-2R) was significantly different between the two groups after treatment for 3 months (P < 0.05). IL-2 associated side effects were rare and mild. CONCLUSION: As an effective and relatively safe biological agent, IL-2 can be added to the standard chemotherapy for pulmonary tuberculosis. PMID- 14521760 TI - [Changes of plasma fibrinolysis system and the effect of captopril in high altitude pulmonary edema]. AB - OBJECTIVE: To investigate the changes of plasma fibrinolysis system and the effect of captopril in patients with high altitude pulmonary edema. METHODS: The plasma levels of tissue plasminogen activator (tPA) and plasminogen activator inhibitor-1 (PAI-1) were measured before and after treatment in the captopril group (group A, 35 cases) and the routine-group (group B, 37 cases) of patients with HAPE, while 20 healthy volunteers served as the control (group C). RESULTS: The plasma level of tPA was (0.40 +/- 0.14) x 10(3) IU/L in group A vs (0.39 +/- 0.19) x 10(3) IU/L in group B before treatment, and (0.58 +/- 0.13) x 10(3) IU/L vs (0.49 +/- 0.16) x 10(3) IU/L after treatment, and (0.59 +/- 0.17) x 10(3) IU/L in group C. The differences were significant both before and after treatment between group A and group B (P < 0.01, P < 0.05). While PAI-1 was (6.6 +/- 1.8) x 10(3) AU/L in group A vs (6.6 +/- 1.6) x 10(3) AU/L in group B before treatment, and (4.9 +/- 1.5) x 10(3) AU/L vs (5.8 +/- 1.7) x 10(3) AU/L after treatment, and (4.9 +/- 1.3) x 10(3) AU/L in group C. The differences were significant both before and after treatment between group A and group B (P < 0.01, P < 0.05). CONCLUSION: Patients with HAPE may present a disturbance of the fibrinolysis system, which could be reversed by captopril. PMID- 14521761 TI - [Association between polymorphisms in the gene coding for glutathione S transferase P1 and chronic obstructive pulmonary disease]. AB - OBJECTIVE: To investigate the association between polymorphisms in the gene coding for glutathione S-transferase P1 (GSTP1) and susceptibility to chronic obstructive pulmonary disease (COPD) in a Chinese population. METHODS: This was a pilot study of the molecular epidemiology in patients with COPD. The research design was a case-control study. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) were performed to genotype GSTP1 polymorphisms in exon 5 in 100 COPD patients and 100 age and sex matched healthy controls. Logistic regression analysis method was used to calculate the odds ratio (OR) and the 95% confidence interval (CI). RESULT: There was no significant difference in GSTP1 polymorphisms adjusted by age, sex, body mass index and smoking between COPD patients and controls. CONCLUSION: The gene polymorphism for GSTP1 was not associated with susceptibility to COPD in the Chinese population. PMID- 14521762 TI - [A case report of cystic fibrosis and review of 16 cases of cystic fibrosis in Chinese patients]. AB - OBJECTIVE: To report a case of cystic fibrosis (CF) in a Chinese and to review the clinical features, gene mutations, diagnosis and outcome of CF in Chinese patients. METHODS: A 14-year-old Chinese girl presenting with recurrent productive cough since birth with parasinusitis and otitis media was confirmed to have CF in Peking University First Hospital. Chest CT scan showed bronchiectasis, more severe in the right upper lobe. Sweat tests were taken three times, and the values of Na(+) and Cl(-) were (126.6 +/- 5.4) mmol/L and (108.9 +/- 3.3) mmol/L, respectively. The examination of the pancreas showed no remarkable cystic changes on CT scan and there was no pancreatic insufficiency. Sixteen patients with CF in Chinese reported from 1974 to 1999 were reviewed. RESULTS: Sixteen of the 17 patients (7 males and 9 females, aged from 6 months to 25 years) had clinical data available for analysis. Eleven of them had died, nine before the age of 13. They all suffered from frequent episodes of pneumonia, while 14 had malnutrition and 4 had jaundice. The diagnostic procedures included clinical features, sweat test and autopsy. Four of them had DNA screened and four kinds of novel mutations in the cystic fibrosis transmembrane conductance regulator gene were found. CONCLUSIONS: Chinese patients with CF show similar clinical manifestations to patients in the European and North American populations, but the CFTR mutation was different. PMID- 14521763 TI - [Clinical analysis of pregnancy in second and third trimesters complicated severe acute respiratory syndrome]. AB - OBJECTIVE: To explore and analyze the clinical manifestation, diagnosis, management and maternal-perinatal prognosis of severe acute respiratory syndrome (SARS)-complicated pregnancy in second and third trimesters. METHODS: Clinical data of 5 inpatients with SARS-complicated pregnancy in second and third trimesters from 4(th) February to 17(th) March 2003 were analyzed retrospectively. RESULTS: Five patients were all primigravida (including 2 twins). Two were infected in second trimester while the other 3 in third trimester with 2 hospital-acquired infections and 3 community-acquired infections. All patients had fever (5/5), 3 chills or rigor, 4 cough; 2 with decreased lymphocyte, 2 decreased platelet, 3 elevated alanine aminotransferase (ALT), 4 hypoalbuminemia, 5 abnormal chest radiographs. All 5 patients were cured with 1 requiring intensive care. 5 neonates including 1 twins have been followed up without evidence of SARS infections up to now. In a twin-pregnancy 1 fetus was lost while the pregnant's situation is stable. CONCLUSIONS: Common diagnostic criteria were Suitable for SARS-complicated pregnancy in second and third trimesters, but attention should be paid to the interaction between SARS and special pathological changes during pregnancy. Patients should be isolated and monitored intensively with timely cesarean section in severe cases, which could significantly decrease the maternal-perinatal mortality. The use of corticosteroids and psychological supports need further study. PMID- 14521765 TI - [Study on the related factors of suicidal ideation in college undergraduates]. AB - OBJECTIVE: To evaluate psychosocial factors and patterns on suicidal ideation of the undergraduates in Shanxi province. METHODS: Four thousand eight hundred and eighty-two undergraduates in Shanxi province were investigated with multistage stratified random clustered samples. Factors associated with suicidal ideation were analyzed with logistic regression and Path analysis by scores of Beck Scale for Suicide Ideation (BSSI), Suicide Attitude Questionnaire (QSA), Adolescent Self-Rate Life Events Check List (ASLEC), DSQ, Social Support Rating Scale, SCL 90, Simple Coping Modes Questionnaire and EPQ. RESULTS: Tendency of psychological disorder was the major factor. Negative life events did not directly affect suicidal ideation, but personality did directly or indirectly affect suicidal ideation through coping and defensive response. CONCLUSIONS: Personality played a stabilized fundamental role while life events were minor but "triggering" agents. Mental disturbance disposition seemed to be the principal factor related to suicidal ideation. Above three factors were intergraded and resulted in suicidal ideation in chorus. PMID- 14521764 TI - [An epidemiological study on stress among urban residents in social transition period]. AB - OBJECTIVE: To explore the stress related problems and risk factors among urban residents in the social transition period. METHODS: Samples including 3 666 subjects came from Hangzhou, Guangzhou, Chongqing and Taiyuan under multi-stage sampling procedure. Data were collected through a household interviewing survey. Stress was measured by the Chinese Perceived Stress Scales (CPSS). Distribution of health risk stress (HRS), high level stress were described along the demographic dimension. Multivariate logistic analysis was used to identify the predictors of HRS. RESULTS: CPSS was found to have good validity and reliability among this study sample. The mean score for the total sample was 24.22 +/- 5.81 and the overall prevalence of HRS was 44.54% (95% CI: 42.90 - 46.12). The difference between male (42.95%) and female (46.30%) was statistically significant (P < 0.05). The rate of HRS decreased with age, with the highest rate (59.29%) among those less than 21 years old. The rate of HRS also decreased as educational level increased and differed significantly by occupation. High school students, people who had never been employed and those who had lost their jobs had the highest rates of HRS. The rate of HRS was also related to marriage status, household's economic level, and the level of the social engagement of the subjects. CONCLUSION: Stress experienced by the urban residents became a serious health problem which requires special attention to be paid to certain subgroups in the population. PMID- 14521766 TI - [The application of Chinese health questionnaire for mental disorder screening in community settings in mainland China]. AB - OBJECTIVE: To examine the applicability of Chinese Health Questionnaire (Taiwan version) (CHQ) originated from General Health Questionnaire (GHQ) for mental disorder screening in community settings in mainland China. METHODS: A pilot study was conducted in Hangzhou (n = 377). Three thousand seven hundred and seven subjects were recruited from four cities to validate the results of the pilot study. Validation of the Screening Questionnaire was analyzed, using Relative Operating Characteristic (ROC) method. RESULTS: Cronbach's alpha coefficients were calculated to be 0.79 for the 12-items and 0.89 for 30-items to CHQ version in the first sample, and 0.74 in the second sample (12-items). Four factors were extracted from the CHQ-30, including somatic symptoms, anxiety and worry, social dysfunction, poor family relationship, and depression. CHQ-12 could be explained by a single factor in both samples. The areas under ROC were 0.80 (95% CI: 0.70 - 0.89) for 12 items and 0.72 (95% CI: 0.62 - 0.82) for 30 items. The sensitivities of CHQ-12 and CHQ-30 were found to be 76.9% and 71.8%, and the specificities were 73.8% and 67.9% with Kappa value 0.44 (P = 0.00) and 0.38 (P = 0.00), respectively. The estimated rates of mental disorder were 18.13% (95% CI: 14.16 - 22.10) by CHQ-12, and 22.80% (95% CI: 18.19 - 27.11) by CHQ-30 in the first sample and the rates were 21.72% (95% CI: 20.39 - 23.05) by CHQ-12 in the second sample. CONCLUSION: CHQ, especially CHQ-12 through slight language modification, could be used for epidemiological studies and on community health care to screen for mental disorder in the mainland of China. PMID- 14521767 TI - [Study on the prevalence and risk factors of "subhealth" status in college and university staff in Guangdong province]. AB - OBJECTIVE: To understand the current status and risk factors of "subhealth" in college and university staff in Guangdong province. METHODS: Eight thousand four hundred and seventeen staff in 19 colleges and universities in Guangdong were investigated through a self-developed questionnaire, and statistically analyzed using chi(2) test, chi(2)(strand) test and odd ratio. Judgement was based on the criteria on "subhealth" through Delphi method. RESULTS: Overall incidence of "subhealth" was 69.18% in 8,417 staff members, and the highest was in the age group of 30 - 40 year olds (totally 79.17%). The prevalence of severe "subhealth" in females was significantly higher than that of males (chi(2) = 14.19, P < 0.01). The main risk factors of "subhealth" were occupational stress, psychological factors, bad habits and behaviors. 44.21% of the 8,417 staff were aware of the terminology "subhealth", and 36.84% thought themselves under "subhealthy" condition. CONCLUSION: The health condition of college and university staff in Guangdong was not satisfactory, thus it was essential to carry out active measures of prevention and intervention among this population. PMID- 14521768 TI - [Study on physical and mental health status and influencing factors of middle aged intellectuals]. AB - OBJECTIVE: To understand physical and mental health status of middle-aged intellectuals and related risk factors on mental health under the present social conditions. METHODS: Stratified-cluster sampling and questionnaire survey were used. One thousand five hundred and three middle-aged intellectuals were investigated by SCL-90 test and influential reasons, using general relative risk model. Effects and interaction modes of risk factors were analyzed. RESULTS: The prevalence rate of middle-aged intellectuals was 241.5 per thousand per two weeks, while the prevalence rate of chronic disease was 306.7 per thousand. 6.9% of the middle-aged intellectuals (male 6.1%, female 7.8%) had mental health problems. Results suggested that the major risk factors were, as listed in a order of their effects on mental health disorders: divorced or losing spouse, exposure to poison and noisiness, lack of hobbies, long working hours, having chronic diseases. CONCLUSION: Physical and mental health status of middle-aged intellectuals were under by many kinds of pressures. Multiple measures should be taken to improve the health status of the middle-aged intellectuals. PMID- 14521769 TI - [A study on the variation of goiter rates, urinary iodine and household salt iodine intake among children in West China]. AB - OBJECTIVE: To study the factors leading to the variation of children's total goiter rates, urine iodine and salt iodine in West China. METHODS: Design effect (DEFF) was used to evaluate the variation of indicators. RESULTS: DEFF of children's total goiter rate was about 3.0 in 9 provinces, and the proportion was 75.0%. The DEFF of urine iodine was 1.0 - 3.5 in 6 provinces, and the proportion was 50.0%. The DEFF of intake rate of qualified iodized salt was over 3.0 in 11 provinces. The DEFF of covering rate of iodized salt was about 3.0 in 4 provinces. CONCLUSION: In order to provide scientific basis for IDD surveillance in China, other than bias due to methods of no-sampling, sample size should be further calculated in West China. Thus the main influencing factors of indicator variation could be measured with scientific and reasonable basis. PMID- 14521770 TI - [Evaluation on the effectiveness for self-management of hypertensive patients in a community]. AB - OBJECTIVE: To evaluate the acceptability of self-management project for patients with hypertension and the short term result, and provide the evidence of preventing and controlling hypertension using chronic disease self-management. METHODS: Voluntary patients with hypertension were grouped into the control and experimental groups after matching age, sex, education levels and the number of chronic diseases between the two groups. Data collected through questionnaire after intervention were compared with the baseline data through analysis of covariance and chi(2) test. RESULTS: Rate of awareness for the diagnostic criteria of hypertension, risk factors and associated diseases increased by 56.4%, 50.4% and 37.6% respectively, and the rate of high salt diet intake, body mass index and the times of visiting doctors decreased by 27.1%, 0.8 kg/m(2) and 2.7 times on average in experimental group, as compared with control group. The health status (energy, fatigue, shortness of breathe, pain) improved (P < 0.05). In the similar comparison, the rate in which the hypertensive patients took antihypertensive medicine increased by 20%, and the scores of cognitive symptoms management and of patient communication with physician increased by 3.9 and 1.1 points respectively. The rate of compliance to medication increased by 21.5% compared with themselves. CONCLUSION: The awareness rate of hypertensive patients on knowledge and on unhealthy life style improved after implementing the self management program. Their emotion, healthy behavior, health status and symptoms related to hypertension improved. Data showed that it was effective for hypertensive patients to use chronic diseases self-management. PMID- 14521771 TI - [The effect of dietary intervention on lipidemia in school-aged children]. AB - OBJECTIVE: To evaluate the effect of dietary intervention on lipidemia in school aged children. METHODS: The levels of serum lipids profile were detected in 316 school children aged 7 - 11, from those the subjects of dietary intervention were selected by total cholesterol level above 4.26 mmol/L or low density lipoprotein cholesterol level above 2.23 mmol/L. The subjects were randomly divided into intervention group (120) and control group (40). Children in intervention group were fed with low-cholesterol and low-saturated fatty acid diet, and the control group with normal diet. The duration of intervention was three months. Before and after the intervention, the food intakes, health-related questionnaire and physical examination (height, weight, skinfolds thickness and so on) in the two groups were studied. RESULTS: Compared with the control group, serum cholesterol levels of children under intervention were not significantly changed (TC: 4.64 vs 4.68 mmol/L, P > 0.05; LDL-C: 2.66 vs 2.62 mmol/L, P > 0.05), but the apoA(1) level increased from 1,378.4 mg/L to 1,441.3 mg/L (P < 0.05). There were no changes for any serum lipids indexes in the control group while the dietary intakes of energy, cholesterol and SFA decreased markedly in the intervention group, with the percent of energy from fat decreased from 40.7% to 31.2% and SFA below to 10% (7.7%). Along with the increase of the scores of knowledge on health among children under intervention, the living and eating habit improved (the total scores increased from 24.6 to 27.4, P < 0.05). The increase of height was not significantly different between the two groups. CONCLUSION: With the family based high-risk intervention strategy on the dietary adjustment, the knowledge on health, living and eating habit could be effectively improved in children with lipidemia. However, further research about the intervention effect on the serum cholesterol levels by strengthening the interventional degree, needs to be further studied. PMID- 14521772 TI - [Principal component analysis and integral methods of cerebral vascular hemodynamic parameters]. AB - OBJECTIVE: To establish a predicting model for stroke according to cerebral vascular hemodynamic indexes and major risk factors of stroke. METHODS: Participants selected from a stroke cohort with 25,355 population in China. The first step was to carry out principal component analysis using CVHI. Logistic regression with principal component and main risk factors of stroke were then served as independent variables and stroke come on as dependent variables. The predictive model was established according to coefficient of regression and probability of each participant was also estimated. Finally, ROC curve was protracted and predictive efficacy was measured. RESULTS: The accumulative contribution rates of four principal components were 58.1%, 79.4%, 88.4% and 94.6% respectively. Seven variables were being selected into the equation with the first to fourth principal component as history of hypertension, age and sex. Area under ROC curve was 0.855 and optimal cut-off point was probability over 0.05. Sensitivity, specificity and accuracy of stroke prediction were 80.7%, 78.5% and 78.5% respectively. CONCLUSION: The model established by principal component and regression could effectively predict the incidence of stroke coming on. PMID- 14521773 TI - [Rapid detection of genotypes of TT virus using a heteroduplex mobility assay]. AB - OBJECTIVE: To establish a simple, sensitive, specific and less-costly method for detecting genotypes of TT virus (TTV). METHODS: TTV DNA was tested by nested polymerase chain reaction (nPCR) in sera from 180 patients with different types of viral hepatitis and 96 normal individuals in Beijing. TTV genotypes were determined in 40 sera collected from TTV DNA positive patients by heteroduplex mobility assay (HMA) and through sequencing. RESULTS: The positive rates of TTV DNA in viral hepatitis patients and normal individuals were 22.2% (40/180) and 19.8% (19/96), respectively (chi(2) = 0.220, P = 0.639). TTV DNA positive rates of patients with hepatitis A, B, C, E and non-A to E were 20.0% (6/30), 16.7% (5/30), 23.3% (7/30), 36.7% (11/30) and 18.3% (11/60), respectively. Of 40 TTV DNA positive patients, 20 (50.0%) were TTV G1, 7 (17.5%) TTV G2, 10 (25.0%) coinfected with different genotypes of TTV, and 3 untyped by HMA. Twenty G1 and 7 G2 detected by HMA were confirmed by sequence analysis. Of 10 patients coinfected with different genotypes of TTV, 5 were G1 and G2, 2 G1 and G3, 1 G1 and G4, 1 G1 and G3, and 1 with G1, G2 and G3 coinfections. CONCLUSION: HMA was recognized as simple, sensitive, specific and less-costly, thus could be used for genotyping of TTV. PMID- 14521774 TI - [Detection of SEN virus (subtype D/H) infection in Shenzhen]. AB - OBJECTIVE: To investigate the prevalence of newly identified single-chain DNA virus (SENV) infection in Shenzhen. METHODS: Nested polymerase chain reaction (nPCR) was established using primers from ORF1 region of SENV genome. Six hundred and one sera samples from different populations were detected for SENV DNA (D and H subtype) by nPCR. Products of PCR were cloned into T-vector and sequenced. RESULTS: The positive rates of SENV DNA in different populations were as followed: 27.8% in patients with hepatitis B, 22.2% in patients with hepatitis C, 26.9% in hemodialysis patients and 39.3% in IDUs. Among blood donors, the positive rates of SENV DNA were 28.1% in unqualified blood donors, 31.3% in blood donors with an elevated ALT levels and 15.1% in qualified blood donors. The infection rates of SENV in unqualified blood donors and blood donors with an elevated ALT levels were obviously higher than in qualified blood donors (chi(2) = 8.29, P < 0.01 and chi(2) = 6.03, P < 0.01). There was a 6.8% difference of nucleotide between SENV-D standard subtype and 6 isolates with 13.5% difference of nucleotide between SENV-H standard subtype and 4 isolates from Shenzhen. CONCLUSION: Results suggested that SENV infection was common in high-risk groups in Shenzhen. PMID- 14521775 TI - [Principal component analysis on ultrasound indexes of schistosomiasis and the assessment on prevalence rate]. AB - OBJECTIVE: To explore the synthetical index for diagnosing schistosomiasis with ultrasound and to assess the prevalence rate with the index. METHODS: Ultrasound indexes of schistosomiasis Japonicum were analyzed by principal component analysis, and the synthetical indexes were assessed by ROC curve. RESULTS: Among the abnormal rates of the 6 indexes, the lowest was 1.6% comparing with the highest of 59.5%. Significant difference was noficed among the abnormal rates (chi(2) = 631.1, P < 0.01). The individual correlation of the six indexes to each other as will as with age distribution was significant (P < 0.05). The three principal components reflected the degree of pathological changes on liver and spleen. The first principal component was the factor reflecting the degree of liver pathological changes, and the second and third principal components reflected the degree of pathological changes on spleen. The synthetical index D(1) = 0.047X(1) + 0.428X(2) + 1.247X(3) + 0.095X(4) + 0.002X(5) + 0.213X(6) - 12.837 was found by adding the three weight principal components, and it's area under the ROC curve was 0.957. When -1.70 was taken as the critical value, the abnormal rate of population was 66.3%, close to the resident's actual prevalence rate 66.9%. CONCLUSION: Ultrasonography was considered as a method which could rapidly assessing the resident's prevalence rate in the endemic areas of schisitosomiasis Japonicum, and could also provide powerful information for development of strategy on chemotherapy. PMID- 14521776 TI - [Study on the genetic association between DRB3 and DRB1 loci in the human MHC region and psychotic symptoms of schizophrenia]. AB - OBJECTIVE: The genomic region of the human major histocompatibility complex (MHC) is located in the short arm of chromosome 6 (6p). Linkage studies have shown that the 6p region may contain a gene for schizophrenia, the MHC region has thus become particularly important in searching for the schizophrenia susceptibility gene. The present study was designed to investigate the genetic association of DRB3 and DRB1 genes with psychotic symptoms of schizophrenia. METHODS: PCR-based restriction fragment length polymorphism (RFLP) analysis was applied to genotype two single nucleotide polymorphisms (SNPs) located in the DRB3 locus and in the DRB1 one in 116 Chinese Han family trios consisting of fathers, mothers and affected offspring with schizophrenia. Chi-square (chi(2)) test and haplotype based relative risk (HRR) analysis were used on genotyping data. RESULTS: Data on HRR analysis did not show a genetic association either between the DRB3 locus and schizophrenia or between the DRB1 locus and the illness. However, the SNP rs707954, a G to T base change, present in the DRB1 locus showed strong association with idea of reference (chi(2) = 5.484, df = 1, P = 0.019), while the genotype of rs707954 showed an association with idea of reference (chi(2) = 6.771, df = 2, P = 0.034) as will as with apathy (chi(2) = 12.110, df = 4, P = 0.017). CONCLUSION: DRB1 locus seemed likely to be associated with psychotic symptoms as idea of reference and apathy. Further studies were necessary to reveal the relations between DRB1 gene or nearby locus with its susceptibility to schizophrenia. PMID- 14521777 TI - [Study on prevalence rates of hypertension and chronic diabetic complications of in-patients with diabetes]. AB - OBJECTIVE: To understand the relationship between prevalence rates of chronic diabetic complications and related macro-vascular diseases and hypertension, to provide evidence for diabetes prevention and control. METHODS: To retrospectively analyze the clinical and laboratory data on 24 496 in-patients with diabetes during Jan. 1, 1991 to Dec. 31, 2000 from 30 provinces, municipalities and autonomous regions. RESULTS: The prevalence rate of hypertension was 31.95%. The prevalence rates of other chronic diabetic complications and related macro vascular diseases were 12.2% for cerebrovascular diseases, 15.9% for cardiovascular diseases, 5.0% for lower extremity vascular diseases, 34.3% for eye diseases, 33.6% for nephropathy and 60.3% for neuropathy, with a total rate of chronic diabetic complications and macro-vascular diseases of 73.2%. Hypertension was related with all kinds of chronic diabetic complications and macro-vascular diseases odds ratio (OR) of related systolic blood pressure was 1.148 - 1.866, and OR related diastolic was 1.141 - 1.242. CONCLUSIONS: The prevalence rates of chronic diabetic complications and related macro-vascular diseases of in-patients with diabetes mellitus were much higher than those of non diabetics, and was close to the levels seen in the western countries. Hypertension, especially high systolic blood pressure was the most important risk factor of chronic diabetic complications and the related macro-vascular diseases. PMID- 14521778 TI - [A cross-sectional study on angiotensin-converting enzyme and angiotensin II type I receptor gene polymorphism and cerebral infarction]. AB - OBJECTIVE: To explore the relation of angiotensin-converting enzyme (ACE) gene polymorphism, angiotensin II type I receptor (ATIR) gene polymorphism and other factors on cerebral infarction. METHODS: One thousand three hundred fifty-one subjects from Tangshan coalmine were enrolled with study method of cluster sampling. Face to face interviews were conducted to fill in questionnaires by trained interviewers. ACE gene, ATIR gene and inflammation factors including tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), IL-8, IL-10, C reactive protein (CRP), fibrinogen (Fg), fibrin monome polymerized velocity (FMPV), absorbance maximum (A(max)), FMPV/A(max), were measured. RESULTS: No different prevalence rates of ACE genotype were found on cerebral infarction. The distributions of AA genotype of ATIR gene in the cerebral infarction was higher than that of the controls. The prevalence of AA genotype was higher than other groups, but the prevalence of combined genotype did not show much difference. Under the existence of factors that related to cerebral infarction, AA genotype frequencies were higher than those of non-smoking and with hypertension. IL-6, ATIR gene polymorphism, sex, FMPV/A(max) were strongly related to cerebral infarction. The level of IL-6 was higher than the normal ones. CONCLUSIONS: The prevalence of cerebral infarction obviously increased in the hypertensive groups having AA genotype of ATIR gene. In the cerebral infarction groups, the level of IL-6 was higher than that in the normal population, indicating that these can be resulted from local inflammation and immunity reactivity. Environmental and genetic factors in the pathogenesis of cerebral infarction might have coordinating functions. PMID- 14521780 TI - [Study on meteorological factors-based neural network model of malaria]. AB - OBJECTIVE: In order to provide reliable data for strategies development on prevention, a meteorological factors-based predicating model for malaria forecast was studied. METHODS: Data on malaria occurrence and climate changes from 1994 to 1999 in counties in Yunnan province was collected and analyzed with software packages of FoxPro 6.0 and Excel 5.0. The forecasting model for malaria occurrence was established, using the Neural Network Toolbox of Matlab 6.1 software package. In the studies of forecasting model, data of malaria and meteorological factors from 1994 to 1999 in Honghe state in Yunnan province was chosen. The meteorological factors included average monthly pressure, air temperature, relative humidity, monthly maximum air temperature, minimum air temperature, rainfall, rainday, evaporation and sunshine hours in the study. The established forecasting model was also tested and verified. RESULTS: The BP network model was established according to data of diseases and meteorological factors from Honghe state in Yunnan province. After training the neural network for 100 times, the error of performance decreased from 3.23608 to 0.035862. Verified by fact data of malaria, the efficiency of malaria forecasting was 84.85%. CONCLUSION: Neural network model was effective for forecasting malaria. It showed advantages as: strong ability for analysis, lower claim for data, convenient and easy to apply etc. Neural network model might be used as a new method for malaria forecasting. PMID- 14521779 TI - [A case-control study on risk factors for nosocomial infection of epilepsy in hospitalized children]. AB - OBJECTIVE: To determine the incidence and risk factors for nosocomial infection in child epilepsy. METHODS: A retrospective study was conducted among all (292 cases) hospitalized children epilepsy patients in First Affiliated Hospital of Fujian Medical University from 1996 to 2000 in Fuzhou city. With all patients with nosocomial infection as cases and all patients without nosocomial infection as controls, a case-control study on risk factors for nosocomial infection was carried out. Available data were analyzed by one-way Chi-square test and unconditional logistic multiple regression model. RESULTS: One hundred fourteen cases of nosocomial infection were identified among 292 cases with epilepsy with an incidence of 39.0% (114/292). The one-way Chi-square test showed that nosocomial infection was significantly associated with age below 3 years (OR = 2.55, P < 0.01), length of hospitalization over 14 days (OR = 4.75, P < 0.01), low intelligence (OR = 3.13, P < 0.01), receiving antibiotic unreasonably (OR = 3.51, P < 0.01), using gastrogarage (OR = 3.12, P < 0.01), other invasive operation (OR = 1.85, P < 0.05) dyskinesia or palsy (OR = 3.51, P < 0.01), and urinary nitrogen beyond normal range (OR = 5.00, P < 0.05), etc. Unconditional logistic multiple regression analysis revealed that the length of hospitalization over 14 days (OR = 4.30, OR 95% CI: 2.48 - 7.46, P < 0.01), taking antibiotic unreasonably (OR = 2.74, OR 95% CI: 1.30 - 5.77, P < 0.01), using gastrogarage (OR = 3.04, OR 95% CI: 1.28 - 7.18, P < 0.05), and low intelligence (OR = 2.32, OR 95% CI: 1.34 - 4.01, P < 0.01) were independent risk factors for nosocomial infection. The tendency chi-square test showed that the longer stay in the hospital with more kinds of antibiotic used and more gastrogarage they used, the greater the risk of nosocomial infection was. CONCLUSION: Data suggested that occurrence of the nosocomial infection of children epilepsy patients was correlated with the length of hospitalization over 14 days, unreasonable using antibiotic, using gastrogarage and low intelligence. PMID- 14521781 TI - [Application of capture-recapture method on injury control]. AB - OBJECTIVE: To explore relationships among several capture-recapture methods to be used in injury studies. METHODS: Comparing the method on the estimation number of missed cases with supergeometric distribution and proportional methods and study the relationship between the three methods. RESULTS: Results from estimation method for number of missed cases and supergeometric distribution were identical while the formula of estimation method for number of missed cases could be induced from one of supergeometric distribution formula. The distribution of injured population did not belong to the negative binomial distribution. The estimation range was not the same between proportional method and supergeometric distribution. CONCLUSIONS: Either supergeometric distribution or estimation method for number of missed cases could be chosen, but the former was simple in calculation. Considering the estimating range was not the same between supergeometric distribution and proportional method, conditions for application must be considered during implementation. PMID- 14521782 TI - [Analysis on Clenbuterol pollution in swine products in Beijing, 2002]. AB - OBJECTIVE: To explore the status of clenbuterol pollution in swine products in Beijing city in 2002. METHODS: European Union method (EUR 15127-EN Cy2.3) was adopted to examine the samples. Samples were screened by enzyme-linked immunosorbent assay (ELISA) and confirmed by GC-MS. Detected limit of the method was 0.5 micro g/kg. Samples, including lung, liver, pork, kidney and urine of swine, were collected from slaughterhouses, refrigeratories and markets in 11 districts of Beijing. RESULTS: The results indicated that 185 out of 1 379 samples were positive with an annual positive rate of 13.4%. The highest was 15.7% in lung of swine, followed by urine 15.2% and pork liver 14.0%. CONCLUSION: Rates of detection had decreased from 30.0% to 2.7% during 2002. PMID- 14521783 TI - [Active surveillance on Listeria monocytogenes in seven kinds of food in 11 provinces of China in 2001]. AB - OBJECTIVE: To carry out national active surveillance on Listeria monocytogenes in foods in China. METHODS: Four thousand and thirty-four random samples from raw meat, meat product, raw milk, vegetable, yoghurt, icecream and aquatic product were collected in 11 provinces (cities), and examined for Listeria monocytogenes according to the national standard method and confirmed by BAX system (DuPont Qualicon, Wilmington, DE). RESULTS: Seventy isolates four kinds of foods in seven provinces were found to have LM according to the national standard method with a total isolate rate of 1.74%. In Fujian, the rate was higher than in the other provinces. Raw meat was found to be most heavily contaminated in seven kinds of foods. Comparing to national standard method, BAX system showed good sensitivity (> 98%) and specificity (> 97%). CONCLUSION: In each province seven kinds of food were all contaminated by Listeria monocytogenes to some degrees, suggesting that local sanitary surveillance should be strengthened. BAX system can be used to correctly and quickly screen Listeria monocytogenes. PMID- 14521784 TI - [Study on of dietary pesticide residues in Chinese residents]. AB - OBJECTIVES: To investigate the change of pesticide residues in Chinese dietary through analysis on results of the pesticide residues in the Chinese total diet study carried out the first in 1990. METHODS: Organochlorine, such as HCH and DDT of 9 groups and 15 organophosphorus pesticide residues of 3 groups in four regions of China were determined by gas chromatography-electron capture detector (GC-ECD) and gas chromatography-flame photometric detector (GC-FPD) respectively. According to the amount of pesticide residues in various foods and the amount of food consumption in different areas, we calculated the amount of dietary intake of pesticide residues, then compared with the acceptable daily intake (ADI) or provisional tolerable daily intake (PTDI). The contaminated samples were validated by gas chromatography-mass spectrometry (GC-MS). RESULTS: Results showed that the total dietary daily intake of HCH per person was decreasing from 5.04 micro g in 1990 to 3.11 micro g in 2000, where as the total dietary daily intake of DDT per person was decreasing from 20.47 micro g in 1990 to 2.15 micro g in 2000. The result was less than 1 percent difference of PTDI (0.01 mg/kg bw) established by the Joint FAO/WHO Meeting on Pesticide Residues (JMPR) in 2000. Compared to the results in 1990, the intake of HCH from animal foods increased a little, due to the contaminated of aquatic products by Lindan in the second Southern region and the first Northern region. None of the organophosphorous pesticide residues analyzed were detected in cereals, vegetables and fruits. CONCLUSION: The intake level of pesticide residues in Chinese dietary in 2000 was considered to be low, and the abuse of organophosphorous pesticide seemed to be under effective control. PMID- 14521785 TI - [The introduction of complementary food of children under the age of three years in Tibet of China]. AB - OBJECTIVE: To understand the feeding status of Tibetan children aged below three years old. METHODS: Cross-sectional study and randomly sampling were used. The information on complementary food was obtained through the recollection from mother of the child. Data of complementary food of Tibet children was compared with those of the children from the surveys of nine cities in China and 40 poor counties in western China. RESULTS: One thousand six hundred and fifty-five children were studied. The findings indicated that the age of introduction of Zanba (a kind food of mixture of highland barley flour and butter) was 1.98 months after birth and the age of introduction of egg and meat was about 8 - 9 months. Boys were fed with Zanba earlier than girls but there was no significant difference in the introduction of other complementary food between sexes. Urban children were introduced with complementary food earlier than rural children, and children in the agricultural region earlier than those in the pasturing region. The proportion of no introduction of any complementary food by the age of 4 months was about 10.6% for urban children, but 21.7% for rural children while 14.2% for children in the agricultural region and 30.9% for children in the pasturing region. The proportion of no introduction of complementary food was 11.5% for Tibetan children compared with 45.2% for the children of 40 poor counties in western China. The proportions of introduction of the following complementary foods were 89.1% for Zanba, 22.1% for egg, 29.1% for fresh milk, 23.5% for meat, 17.5% for vegetable, 0.75% for bean products, 1.03% for fish and 3.35% for chicken respectively. CONCLUSIONS: The main complementary foods for Tibetan children included Zanba, milk, mutton and beef. There was a small amount of vegetable in diet. The introduction of bean, chicken and fish was not common. Early and monotony seemed to be the character of introduction of complementary food for Tibetan children. Improving introduction of complementary food is important in enhancing the nutritional status of Tibetan children. PMID- 14521786 TI - [Investigation on reproductive tract infection among floating married women of childbearing age in Fengtai district in Beijing]. AB - OBJECTIVE: In order to develop plans for effective intervention measures, prevalence and health-seeking behavior related to reproductive tract infection among floating married women of childbearing age in Fengtai district in Beijing were studied. METHODS: Cross-sectional study was carried out. Two thousand and sixty-nine eligible women were randomly selected from strata based on their home provinces. From June to July 2001, the subjects were given face-to-face interview at the Fengtai family planning clinic in Beijing using standard questionnaire followed by gynecologic examination and laboratory tests. RESULTS: Thirty point three percent of the subjects were found to have reproductive tract infections (RTI) by laboratory tests. Prevalence rates of bacterial vaginosis, candida and trichomonas vaginitis were 22.2%, 4.9% and 2.1% respectively. Prevalence rates of chlamydia, gonorrhea, condyloma acuminatum and syphilis were 2.2%, 1.6%, 0.5% and 0.2% respectively. Of these infected women, only 43.1% (270/626) were symptomatic, and 61.5% (166/270) of these women with symptoms had sought treatment. CONCLUSION: Compared to other results in the literature, we found a relatively high prevalence of RTI in our study population. Only a small proportion of these infected women were symptomatic but only few of them sought treatment. We suggested that the provision of more family planning service and promotion of RTI knowledge to the floating women of childbearing age. PMID- 14521787 TI - [A cross-sectional study on injuries in residents at the community level of Zhejiang]. AB - OBJECTIVE: To study the incidence of injuries among residents living in the communities in Zhejiang. METHODS: A household questionnaire survey was conducted to 16 899 residents in four communities. RESULTS: The incidence of injuries was 16.11% in total, higher in urban area than in rural area and higher in men than in women. The leading causes of injuries were falls (4.80%), knife-cutting or by sharp articles (3.86%), traffic accident (2.07%), collision (1.99%), scalds (1.13%) and animal bites (1.06%). Peak incidence of age group was different in different injuries, such as traffic accident was high in 45 - 60 age group and falls was high in 5 and 60 age group. Direct economic loss for each case with injury would cost 324.21 RMB yuan. CONCLUSION: Injury was common and frequently occurred among residents at the community level which seriously on danger their health and families. PMID- 14521788 TI - [A prospective cohort study on injuries among school-age children with and without behavior problems]. AB - OBJECTIVE: To study the annual incidence of injuries and the relationship between behavior problems and injuries among school-age children. METHODS: A prospective cohort study on injuries for 1-year follow-up period was conducted among 2 005 school-age children selected by cluster sampling from three primary schools in Maanshan city. They subjects were divided into two groups with or without exposure according to behavior problems rated by the Rutter Child Behavior Questionnaire at the beginning of the study. Nonparametric test was performed to analyze the differences in injuries between the two groups of children, and the influential factors for injuries were analyzed with multi-classification ordinal response variable logistic regression model. RESULTS: The overall incidence rate for injuries in school-age children was 42.51%, while among children with and without behavior problems were 64.87% and 38.85%, respectively. There were significant differences between the two groups (u = -6.054, P = 0.000). However, the incidence rates of injuries in school-age children with antisocial (A) behavior, neurotic (N) behavior and mixed (M) behavior were 66.99%, 67.41% and 61.40%, respectively. No significant differences were found among them (u(A,N) = 0.052, P = 0.958; u(A,M) = -0.400, P = 0.689; u(N,M) = -0.364, P = 0.716). Multivariate analysis indicated that injuries in school-age children were associated with children behavior problems, maternal age at childbirth, bad conditions during mother pregnancy, education background of mother, prevention measures for safety at home and the child accompanied to travel between school and home by adults. Behavior problems of children seemed to be the major risk factors for injuries. CONCLUSION: Children with behavior problems represented a significant risk group for injuries among school-age children. When planning intervention strategies on injuries, behavior problems should be emphasized to ensure optimal effectiveness of intervention. PMID- 14521789 TI - [Study on hospitalization expenses of flood disaster areas' residents of Dongting Lake in Hunan province in 1998]. AB - OBJECTIVE: To study the expenses of hospitalization among the population in the flood disaster areas of Dongting Lake in Hunan province in 1998. METHODS: Descriptive epidemiologic study were conducted to analyze hospitalization expenses of the residents of 55 villages in flood disaster areas in 1998; single factors analysis and logarithmic linear regression analysis were carried out to explore influencing factors about hospitalization expenses of the residents. RESULTS: The hospitalization rate was 4.59% with an average hospitalization expenses of 667.42 Yuan in the flood disaster areas' residents of Dongting Lake in 1998. Compared with populations without suffering from flood, hospitalization rate and the average hospitalization expenses of flood disaster Areas' residents of Dongting Lake in 1998 were higher and had significant difference. The average hospitalization expenses in 1998 was affected by flood types, family income, gender, age, literacy, occupation, outcome after leaving the hospital and hospital ranks. CONCLUSION: These results implied that the flood disease aggravated inhabitants' burden of disease in Dongting Lake areas; the factors influencing the average hospitalization expenses were multiple, and synthetic measures should be taken in the prevention and control of flood disaster. PMID- 14521790 TI - [Surveillance on hemorrhagic fever with renal syndrome in Guizhou during 1984 2000]. AB - OBJECTIVE: To explore the relations between different areas on hemorrhagic fever with renal syndrome (HFRS) in Guizhou. METHODS: Various forms of infectious areas were surveyed continuously through epidemiologic surveillance system. RESULTS: In fixed areas under surveillance system, the mean positive rate of HFRSV among Apodemus agrarius was 3.39%, comparing with Rattus norvegicus 1.61% in Apodemus infectious areas of Zunyi county, 3.19% in Rattus norvegicus, but no HFRSV of Apodemus agrarius was identified in Rattus infectious area of Shiqian county. Both Apodemus and Rattus infectious areas were relatively stabilized. In both banks of Luowang river, Kaiyang county, which had been identified as areas of infections for Apodemus in the eastern part, Rattus infectious area in the west, slow change was noticed. In 1983 - 1984 was not found in Apodemus agrarius HFRSV, however the infectious rate of HFRSV in Apodemus agrarius was 13.85% (Ag 1/65, Ab 8/65) in the western part of the province in 1995 - 1998. CONCLUSION: Both Apodemus and Rattus infectious areas were stabilized but changed slowly. Mixed type and the result of mutual penetration were noticed. PMID- 14521791 TI - [Study on the immuno-effects of recombinant hepatitis B vaccination in adults]. AB - OBJECTIVE: To evaluate the immuno-effects of hepatitis B (HB) vaccination in adults. METHODS: Five groups were sampled by means of cluster sampling, and serum HBsAg, anti-HBs and anti-HBc were tested in every group at people aged from 18 to 50. Recombinant HB vaccine was injected to the ones that HBsAg, anti-HBs and anti HBc were all negative. Concentration of anti-HBs in serum was tested after one year and three years of vaccination. Immuno-effects of recombinant HB vaccination in adults at different ages and between sexes, were then calculated. RESULTS: Good immuno-effects of recombinant HB vaccination in adults were noticed. After one year and three years of vaccination with 5 micro g recombinant HB vaccine, the anti-HBs positive rates were 82.76%, 70.77% while the serum concentrations of anti-HBs were 55.91 mIU/ml and 35.41 mIU/ml respectively. When 10 micro g was used, the concentrations were 83.74%, 72.22%, 56.89 mIU/ml and 30.29 mIU/ml respectively. The effects did not show significant differences between different doses on 10 micro g and of 5 micro g. Concentration of anti-HBs reduced when time went by. The factors such as age and sex influenced the effects of immunity on recombinant HB vaccination. CONCLUSION: Good immunity could be obtained when recombinant hepatitis B was vaccinated in vulnerable population aged 18 to 50. PMID- 14521792 TI - [Study on spotted fever group Rickettsiae in Guangdong province]. AB - OBJECTIVE: To explore the existence of spotted fever group Rickettsiae (SFGR) in Guangdong province. METHODS: Sera were tested to find the SFGR in population and host animals. The target samples were screened by polymerase chain reaction (PCR), and Rickettsiae was isolated with embryonated hen eggs and identified by serological tests. RESULTS: Eight hundred and sixty people in natural condition and 321 of mice were determined. The mean positive rate of healthy population was 3.84%. To compare results among elected places, Fisher's exact test was applied. The difference was suggestive (P < 0.01), and there was no significant difference between mountain and plain areas. There was also no significant difference between mountain and plain areas (P > 0.05). Positive rate of mice was 4.67%, with Rattus fulvescens, Rattus edwardsi, Bandicota indica 11.59%, 12.90%, 3.13% respectively. It was the first time that SFGR antibodies in Rattus fulvescens, Rattus edwardsi, Bandicota indica were reported. A total number of 321 mice spleens and 394 ticks from the surface of mice body were collected. Two strains of SFGR, GDFK58-2000 and GDFK59-2000, were isolated in the ticks from the body surface of 2 Rattus fulvescens. They were identified as Rickettsia sibirica by serological tests. Five hundred thirty-three bp OmpA gene fragments of the two strains were cloned and sequenced. Compared with other relevant strains in Genbank, the rates of homology of nucleotide sequences of GDFK58-2000 and GDFK59 2000 and other Rickettsia sibirica strains were from 99.6% to 100%, and the homology of amino acid speculated was 100%. CONCLUSION: It has been proved that epidemic areas of north Asia tick-transmitted SFGR, did exist in Guangdong province confirmed by hostanimals, transmission vectors and aetiology. PMID- 14521793 TI - [Mutation of envelop protein V3 loop in HIV-1 epidemic in Liaoning province]. AB - OBJECTIVE: To study the characteristics of human immunodeficiency virus (HIV-1) V3 loop amino acid mutations among HIV-1 infected people in Liaoning province. METHODS: The whole blood samples of the HIV carriers and AIDS patients were collected in Liaoning province, China and were extracted PBMC genome DNA. HIV-1 V3 and flanking region sequences were amplified by nest-polymerase chain reaction (nest-PCR) with env specific primers: ED5/ED12 and ED31/ED33. Products were sequenced directly and sequences were aligned, translated and analyzed. RESULTS: In AIDS group, some amino acid mutations at specific position of V3 loop: S to R at position 11, H to S, T and N at position 13, A to V at position 19, F to Y at position 20, Q or D to N at position 25 and 29, were found and all common mutations were associated with T tropic/SI phenotype. The frequency of such amino acid mutations in specific positions was higher in AIDS group than that of the asymptomatic infection group (P < 0.05). In addition, we found some unusual tetramer compositions on the tip of V3 loop: GQGR, APGR and RPGA, GLGR, RPGA in addition to some rare mutations, such as: N to H at position 5 and H to S, F at position 34. CONCLUSION: The amino acid mutations on the V3 loop of HIV-1 epidemic in Liaoning province were in agreement with the results of subtype B, but we observed some rare mutations and unusual tetramer compositions on the tip of V3 loop. PMID- 14521794 TI - [A primary investigation on disseminated histoplasmosis in Hubei]. AB - OBJECTIVE: To investigate the epidemiologic features of disseminated histoplasmosis (PDH) in Hubei province. METHODS: Bone marrow smears of 12 patients diagnosed as Kala-azer in Hubei province including 4 patients in Jingsan, 2 patients in Shashi and each 1 in Yichang, Jinmen, Zhongxiang, Luotian, Xianning and Guanghua respectively were re-examed under microscope. Peripheral blood and bone marrow smears of several patients were detected. After inoculated the bone marrow, peripheral blood, liver and spleen tissue of patients in MLI, the single colony was trans-inoculated in BHIB, SDA and CMA and incubated at 25 degrees C and 35 degrees C. Bone marrow, peripheral blood and bacterial fluid of yeast-phase Histoplasma capsulatum (H.cap) were injected into the abdominal cavity of Kunming mice and nude mice. When symptoms and signs developed, the spleen tissue was separated, then observed under microscope and cultured. Mycelium-phase and Yeast-phase H.cap were inoculated in urase and gelatin medium, then incubated at 25 degrees C and 35 degrees C. Histoplasmin was injected subcutaneously into patients, and then followed for 48 - 72 hours. Amphotericin B was selected to treat the PDH patients. RESULTS: Moriform cell cluster and sausage-shaped cell were not observed in mononuclear-macrophages in the bone marrow smears from 12 patients. Leishman-Donovan body was found only in one patient. There wasn't kinetoplast in the cellular plasm of spores in 11 patients and no transeptae was found. The reaction of H.cap to urease was positive and H.cap did not liquefy the gelatin. It appeared to be mycelium-phase at 25 degrees C but no penicillus and catenulate conidia was found. The characteristic denticle macroconidia was observed but produced red coloring matter. It also appeared to be yeast-phase at 35 degrees C. Yeast-phase spores were observed under microscope. No sausage-shaped spore and transeptae were identified. H.cap could be acquired in the spleen tissue in Kunming mice and nude mice. Bacterium forms, characteristics under microscope and biochemical reaction of mycelium-phase and yeast-phase H.cap were different from some other kinds of dimorphic fungi such as Penicillium marneffei and Histoplasm duboisii etc. CONCLUSION: There were scattered epidemics of PDH in Hubei province. The detection rate of PDH was higher in the southeast area then in the northwest area. The golden standards of clinic diagnosis were mycological culture and inoculation to animals. Amphotericin B was necommended as the first choice for treatment. PMID- 14521795 TI - [Meta-analysis on the association of AGT M235T polymorphism and essential hypertension in Chinese population]. AB - OBJECTIVE: To study on the association of M235T polymorphism of the angiotensinogen gene extron 2 (AGT/M235T) and essential hypertension (EH) in Chinese population by means of Meta-analysis. METHODS: Odds ratios (OR) of AGT M235T genotype distributions in EH patients against healthy controls were analyzed. All the relevant studies were identified, poor-qualified studies eliminated, and the risk of publication bias excluded. The Meta-analysis software, REVMAN4.1, was applied for investigating heterogeneity among individual studies and summarizing the effects across studies. RESULTS: A total of 853 cases and 835 controls from 10 studies were included. No heterogeneity among the studies was noticed. The frequencies of the AGT TT, MT and MM genotypes were 65%, 30%, and 4.9% in cases and 50.6%, 41.8% and 7.5% in controls respectively. The frequencies of the AGT T allele were 80% in cases and 72% in controls. The pooled OR (with 95% CI) of TT vs MT + MM was 1.76 (1.44 - 2.16) (P < 0.000 01) with T vs M 1.54 (1.31 - 1.81). The pooled OR of MM vs MT + TT was 0.67 (0.45 - 1.00) (P = 0.05). CONCLUSION: In Chinese population (mainly the Hans), TT genotype might be associated with the increased risk of EH while MM genotype be associated with low risk of EH. PMID- 14521796 TI - [Factors influencing the diagnose on causes of dyspepsia in central area of Shaanxi province]. AB - OBJECTIVE: To investigate the proportions of functional dyspepsia (FD) and structural diseases within the dyspeptic outpatients in hospitals of different ranking in central area Shaanxi province, and to explore the safety in adopting "symptom and treatment" algorithm used in Western countries. METHODS: A clinical epidemiology survey was carried out by means of a stratified sample of 3 019 dyspeptic outpatients through standardized questionnaire. All of the patients were followed for 4 - 24 weeks, and finally received their diagnoses through a consistent criteria. RESULTS: Proportionally, FD in all the outpatients took up 44.8% with 44.7% benign organic causes and 10.5% malignant diseases. In the patients who had marked alarm symptoms, the proportion of benign and malignant diseases rose to 52.1% and 29.2%, respectively. They were significantly higher than those without alarm symptoms (39.2%, 2.9%) (P < 0.01). All of the dyspeptic patients were divided into 7 groups according to different ages. 64.5% of FD patients were younger than 25 years and the rate of FD declined with age. It was found that only four patients below 35 years old had malignance. The number of cases increased significantly in age 35 - 45 group and reached 30.8% in 65 - 74 group. CONCLUSION: There were some differences noticed within dyspeptic patterns between local area in China and Western countries, and the "symptom and treatment" approach was not entirely suitable to the local area in China. PMID- 14521797 TI - [A study on genetic epidemiology of cerebral infarction]. AB - OBJECTIVE: To explore the genetic model of cerebral infarction and to examine the relationship between cerebral infarction and genetic factor. METHODS: A matched case-control study including 112 pairs pedigrees was carried out. Using the Li Mantel-Gart method and the Falconer method to estimate the segregation ratio and the heritability of cerebral infarction. RESULTS: The prevalence rate of the first-degree and the second-degree relatives in cases was significantly higher than that in controls. The segregation ratio was 0.143 7 (0.113 0 - 0.174 4), significantly lower than 0.25, which showed that cerebral infarction did not possess the characteristics of monogenetic model. The results showed that the heritability of the first-degree and the second-degree relatives were 47.84% and 40.95% higher in male's than in female's. CONCLUSION: Cerebral infarction was a polygenetic disease, and the genetic factors played an important role in the occurrence of cerebral infarction, especially in males. PMID- 14521798 TI - [Retrospective studies on 713 cases chronic obstructive pulmonary disease]. AB - OBJECTIVE: To study the diagnostic criteria of severity of chronic obstructive pulmonary diseases (COPD), early intervention, and the effect of health education on the relationship between smoking and health. METHODS: Data from 713 remissive COPD patients who had results of pulmonary function test in the past 10 years were reviewed. All patients were classified by correlative diagnostic criteria, and the results were compared with the diagnostic criteria of WHO Global Initiative for chronic obstructive lung disease (GOLD). Other indexes such as history of smoking, quantity of smoking and the rate of giving up smoking were also analyzed. RESULTS: According to the diagnostic criteria set by the Chinese Respiratory College in 1997 and traditional pulmonary function criteria, 339 patients were identified as having severe COPD, taking up 56.0% of all COPD patients; 200 patients were moderate (28.0%); 114 patients were mild (16.0%). FEV(1)/FVC in mild COPD patients was 70.5% +/- 1.1%, higher than the criteria of 70% in GOLD. Three hundred and thirty-five patients were smokers, taking up 46.9% of total patients. In these patients, 312 patients were men (93%) and 23 patients were women (7%). The average smoking history was 32.9 years, and the average quantity of daily smoking was 22. Two hundred sixteen patients had given up smoking which took up 64.5% of all the smokers. CONCLUSIONS: It was debatable that FEV(1)/FVC was decisive criteria for COPD diagnoses by GOLD and the revised diagnostic criteria of Chinese Respiratory College in 2002. It might miss diagnosing a number of COPD patients who having combined ventilatory disturbance or were in earlier stages. Only 15.9% of mild COPD patients had seen a doctor, that indicated a very low rate of early intervention which was disadvantageous for the prevention and cure of COPD. In contrast to the rate of 28.0% when giving up smoking in ordinary people who had received general intervention, the rate of giving up smoking in COPD patients remained 64.5% much higher and was dissatisfactory. Health education should be improved to include measures to give up smoking. PMID- 14521799 TI - [Exact logistic regression and its performance to SAS system]. AB - OBJECTIVE: To explore the feasibility of exact logistic regression, used as a complemental method for the maximum liklihood estimation, and to analyse with data small sample, unbalanced structure and highly stratal nature under the situations of questionable results or inexistence of the maximum likelihood estimation. METHODS: Data from 37 postoperative breast cancer cases were analyzed in 1997 by exact logistic regression under SAS system. RESULTS: Data calculated by SAS software showed that Quasi-complete separation of data points was detected but the results of maximum likelihood estimation did not exist, SAS outputs conflicted the results of the last maximum likelihood iteration (likelihood Chi square and score Chi-square have similar P, less than 0.05, but the Wald chi square had a larger P, more than 0.05). Under conditional exact parameter estimation, it appeared that: (1) the joint effect of conditional score statistics was 21.12 with P = 0.000 3; (2) for individual parameters, the effect conditional score statistics of histological classification (grades) was 5.80 with P = 0.020 8; axillary node metastasis (diversion) was 5.74 with P = 0.019 5; tumor size (size) was 0.79, with P = 0.647 6. The effects of tumor histological classification and axillary node metastasis were statistically significant on breast cancer tumour. CONCLUSION: Exact logistic regression seemed to be a very useful method in analyzing data from small sample when the maximum likelihood estimation was either with no effect or did not exist. PMID- 14521800 TI - [Estimating family correlation of quantitative traits using generalized estimating equation]. AB - OBJECTIVE: To study the method for measuring familial correlations of quantitative trait and analyzing family data set of body height. METHODS: Generalized estimating equation 2 (GEE2) was employed to estimating both regression coefficients and the familial correlation. Analyses was carried out on software MAREG. A example from height pedigrees illustrated the method. RESULTS: GEE2 provided robust estimations of regression coefficients and familial correlations simultaneously. In body height the correlations between parents and offspring (r = 0.459) and between siblings (r = 0.671) were significantly higher than those between two parents (r = 0.184) after adjusting gender, residence and birth age. Of the same types of relative pairs, the correlation between pairs with individuals of the same gender (eg. father-son r = 0.603, mother-daughter r = 0.456, male sibling r = 0.947, female sibling r = 0.681) was higher than those individuals of different gender (eg father-daughter r = 0.431, mother-son r = 0.364, sibling with different gender r = 0.530). CONCLUSION: GEE2 should be considered a standard method for the investigation of familial aggregation due to its flexibility and robustness. PMID- 14521801 TI - Surgical staging of cancer: impact on prognosis and potential for bias in clinical trials. PMID- 14521802 TI - Intraperitoneal chemotherapy for ovarian cancer. AB - Intraperitoneal chemotherapy provides a means by which high concentrations of drugs and long durations of tissue exposure can be attained at the peritoneal surface. It has been studied widely in ovarian cancer, a disease in which intra abdominal progression remains the major source of morbidity and mortality. Three large randomized trials have shown improved survival in optimally debulked patients who were treated with intraperitoneal chemotherapy as part of a front line regimen, yet it has not become part of usual therapy. Several factors have contributed to the reluctance to adopt intraperitoneal therapy, including technical issues related to drug delivery and the fact that all of the large randomized trials employed intraperitoneal cisplatin, which has more toxicity than intravenous carboplatin, the current standard of care. Future research is needed for further definition of the clinical benefit of intraperitoneal chemotherapy, modification of existing regimens to minimize side effects, and exploration of intraperitoneal biologic, immunologic, and gene therapy techniques. PMID- 14521803 TI - Consolidation/maintenance chemotherapy for ovarian cancer. AB - A recently reported randomized trial has demonstrated that administration of single-agent paclitaxel to women with advanced ovarian cancer who attain a clinically defined complete response to platinum/paclitaxel-based chemotherapy can substantially improve progression-free survival. Whether this strategy will improve overall survival in this clinical setting remains uncertain. PMID- 14521805 TI - Current research directions for locally advanced cervix cancer. AB - Chemoradiotherapy with either weekly doses of cisplatin or the combination of 5 fluorouracil and cisplatin has become the standard of therapy for patients with locally advanced cervix cancer. Recently reported studies that are directed at improvements in radiation treatment planning, improved tissue oxygenation, and neoadjuvant chemotherapy are likely to lead to improvements in therapy. Nonsurgical preoperative staging with positron emission tomography and sentinel lymph node evaluation could result in effective surgical staging, or even nonsurgical staging, more accurate radiation treatment planning, and improved local and systemic control with a reduction in short-term and long-term morbidity in the treatment of locally advanced cervix cancer. Well-designed, randomized, prospective clinical trials are needed for a more thorough evaluation of these preliminary findings and to set research directions for the next several years. PMID- 14521804 TI - Biologic directed therapies in gynecologic oncology. AB - Evaluation of the genome has provided information concerning the origins of many human diseases, including cancer. Identification of the genes and their protein products has greatly increased our understanding of this complex disease. A variety of cellular processes and pathways stimulate cellular proliferation or inhibit cell death. Many of these pathways are targets for novel therapeutic agents. These agents will usher in a new era of biologically targeted therapeutics. In gynecologic oncology, we are just beginning to investigate these new biologic agents. An appreciation and understanding of these pathways of growth deregulation in gynecologic cancers provide an opportunity for many clinically relevant therapies. This review summarizes the emerging biologic therapies with an emphasis on their relevance to gynecologic malignancies. PMID- 14521806 TI - The role of interval debulking surgery in ovarian cancer. AB - The mainstay of treatment for advanced ovarian cancer is the multimodality approach of debulking surgery and paclitaxel--platinum chemotherapy. The size of residual lesions after primary surgery remains the most important prognostic factor for survival. Optimal primary debulking surgery can be performed in approximately 40% of patients and up to 80% if it is done by gynecologic oncologists, but sometimes at the cost of considerable morbidity and even mortality. Based on a trial conducted by the European Organization for Research and Treatment of Cancer, optimal as well as suboptimal interval debulking surgery increases overall (P=0.0032) and progression-free survival (P=0.0055). However, not all patients who have undergone suboptimal primary debulking surgery seem to benefit from interval debulking surgery. Preliminary data from the Gynecologic Oncology Group interval debulking study (GOG-152) indicate that, if the gynecologic oncologist makes a maximal effort to resect the tumor, patients who have undergone suboptimal debulking surgery probably gain little benefit from interval debulking surgery. PMID- 14521807 TI - Surgery for elderly patients with ovarian cancer. AB - The stage and tumor remnants after radical surgery are the most important prognostic factors in patients with ovarian cancer. Because the incidence of ovarian cancer is highest in elderly women, and mostly advanced stages occur, radical surgery is necessary in this population as well as in younger patients. Age and the surgeon's expertise are the factors that influence the quality of surgical treatment of ovarian cancer. Surgical treatment of ovarian cancer should therefore be confined to centers with the required infrastructure and expertise. PMID- 14521809 TI - Prognostic factors in pediatric Hodgkin disease. AB - The sensitivity of pediatric Hodgkin disease to radiation and chemotherapy has resulted in cure for most children and adolescents who have been diagnosed in the past three decades. Identification of prognostic factors in clinical trials has allowed for tailoring of therapeutic approaches to improve outcome in sequential trials. Tumor burden, symptoms, clinical features, pathology, response to therapy, biology, and host factors are reviewed in this context. New developments should be directed toward identification of factors associated with biologic mechanisms of disease to facilitate the development of biologically targeted therapies that will be more efficacious and less toxic. PMID- 14521808 TI - Prognostic factors and risk-based therapy in pediatric acute myeloid leukemia. AB - Acute myelocytic leukemia (AML) is a heterogeneous disease that accounts for nearly a quarter of the pediatric leukemias. Despite near myeloablative therapy, half of patients relapse and die from their disease. Identification of patients at high risk of relapse early in the course of treatment may allow for treatment modification to improve their outcome. In addition, patients at lower risk of relapse may benefit from treatment de-escalation, sparing them adverse side effects. This review describes prognostic factors that play a major role in the outcome of children with AML and their potential use for treatment stratification in pediatric AML trials. PMID- 14521810 TI - The development of VAC chemotherapy in rhabdomyosarcoma: what does one do for an encore? AB - The establishment of the Intergroup Rhabdomyosarcoma Study (IRS) Committee in 1972 by three extant American pediatric cooperative groups initiated five generations of studies in the multidisciplinary treatment of soft-tissue sarcomas. Taking the lead from the use of vincristine, actinomycin-D, and cyclophosphamide (VAC) by Dr. Ruth Heyn, Dr. Harold Mauer and colleagues developed a more robust VAC as well as three similar drug regimens with hematopoietic growth factor support, which, in the context of surgery with or without radiotherapy, achieved a 3-year failure-free survival rate of 83% (IRS IV) in nonmetastatic embryonal rhabdomyosarcoma. This review documents the development of VAC and some of the new directions currently being pursued by the Children's Oncology Group Soft Tissue Committee. PMID- 14521820 TI - JNC 7 challenge. PMID- 14521821 TI - Summary of the JNC 7 guidelines for the prevention and treatment of high blood pressure. PMID- 14521811 TI - Juvenile myelomonocytic leukemia. AB - Juvenile myelomonocytic leukemia is an aggressive neoplasia of early childhood. Only allogeneic stem cell transplantation (SCT) offers long-term cure. In the absence of an HLA-matched family donor, early SCT from an unrelated donor is the treatment of choice for most children. With clear evidence of a graft-versus leukemia effect and a high post-transplant relapse rate, the outcome of SCT depends, in part, on the management of immunosuppression during the procedure. The impact of pretransplant cytoreductive treatment, such as intensive chemotherapy, splenectomy, or 13-cis retinoic acid, is unclear. Hypersensitivity for granulocyte-macrophage colony-stimulating factor and pathologic activation of the Ras/MAPK pathway play an important role in the pathophysiology of juvenile myelomonocytic leukemia and provide the opportunity for several novel therapeutic approaches. PMID- 14521822 TI - Impact of breast-feeding promotion on infant feeding in the Czech Republic. AB - OBJECTIVES: To assess infant feeding practices, identify factors that influence the duration of exclusive breast-feeding, and evaluate the impact of the National Breastfeeding Promotion Programme in the Czech Republic. HYPOTHESIS: We predicted the positive effect of the program in terms of increased prevalence of breast feeding at discharge from the hospital and 6 months later. DESIGN: Multicenter cross-sectional survey. Mothers were randomly selected and interviewed in maternity hospitals in 1998 and were reinterviewed 6 months later. SETTINGS: Nine maternity hospitals in 6 cities across the country and households. PARTICIPANTS: 1104 mothers who gave birth within the 38th to the 42nd week of pregnancy to a healthy child with a minimum birthweight 2500 g; 1019 mothers were reinterviewed 6 months later. INTERVENTION: The National Breastfeeding Promotion Programme implemented since 1991. VARIABLES MEASURED: Breast-feeding rates at discharge from the hospital and 6 months later, participation in prenatal classes, strategies related to breast-feeding promotion in the hospital, and support following discharge. ANALYSIS: Epi Info 6, SPSS (analysis of variance, F test), chi2. RESULTS: On leaving the hospital, 93.5% of newborns were exclusively breast fed. Six months later, 23.1% of infants were breast-fed exclusively and 29.9% of infants were breast-fed while receiving complementary food. Participation in prenatal classes, first suckling within 2 hours of birth, breast-feeding on demand in the hospital, exclusive breast-feeding on leaving the hospital, and the duration of breast-feeding recommended by a pediatrician were positively related to the duration of exclusive breast-feeding (P <.001). CONCLUSIONS AND IMPLICATIONS: Compared with national data from the last 2 decades, the data from this study proved the increasing rates of breast-feeding at discharge from the hospital and 6 months later. The findings indicate the effectiveness of the National Breastfeeding Promotion Programme. However, further implementation of breast-feeding promotion strategies in health facilities coordinated by the Ministry of Health is needed. PMID- 14521823 TI - Body image, weight, and food choices of Latina women and their young children. AB - OBJECTIVE: To investigate body image perceptions of women about themselves and their young children and their relationship to their food choices and those of their children. DESIGN: Descriptive and correlational study. PARTICIPANTS AND SETTING: 187 low-income, Latina women and their children, ages 5 to 7 years and 52% female, in New York City. VARIABLES MEASURED: Body image, food frequency, body mass index (BMI) of mothers and children, and food choice criteria of mothers for their children. ANALYSIS: Descriptive statistics and correlations. RESULTS: All of the women selected a relatively thin body image as the most desirable, attractive, fit, and healthy (about 2.5 on a scale of 1-7). Body size dissatisfaction or wish to be thinner was significantly associated with more healthful diets. Tertiles (thirds) of children at the 50th and 75th mean BMI-for age percentiles were thought to be too thin to be attractive or healthy and the third of children with a mean above the 97th percentile only barely too large. Mothers with the highest body mass indices may make the least healthful choices for their children. CONCLUSIONS AND IMPLICATIONS: These Latina women preferred a thin figure for themselves but a plumper figure for their children. Culturally competent nutrition education incorporating body image issues needs to assist mothers in understanding the health consequences of childhood obesity, recognizing when their children are overweight, and understanding the importance of healthful food choices for their children. PMID- 14521824 TI - Public versus private food assistance: barriers to participation differ by age and ethnicity. AB - OBJECTIVE: To examine participation in the Food Stamp Program, food pantries, and soup kitchens and to identify reasons food-insecure households choose not to participate. DESIGN: Cross-sectional retrospective cohort study. SETTING: In respondents' homes. PARTICIPANTS: 330 randomly selected low-income households (below 185% of poverty). MAIN OUTCOME MEASURES: Participation in any of 3 public or private food assistance programs and barriers to participation in each program. ANALYSIS: Chi-square tests of association between program participation and sociodemographic characteristics. Logistic regression tested for associations between program participation and ethnicity and between food security status and household composition while controlling for potential confounding factors. RESULTS: Controlling for socioeconomic status, Black households are less than half as likely to receive food stamps (odds ratio [OR] = 0.49; P <.02) as Hispanic households. Hispanic households are more likely than non-Hispanic households to say that they feel uncomfortable using food pantries (P <.01). Elderly households are less than half as likely to receive food stamps (OR = 0.44; P =.04) as nonelderly households and are more likely to say that they feel uncomfortable receiving food stamps (P =.05). CONCLUSIONS AND IMPLICATIONS: Low income households' perceptions of which programs are socially acceptable differ by race and age. Outreach for food stamps and private food assistance should accommodate these differences so that food-insecure households can benefit from all available food assistance. PMID- 14521825 TI - A test of conversational and testimonial messages versus didactic presentations of nutrition information. AB - OBJECTIVE: Conversational and testimonial approaches to presenting messages relevant to fruit and vegetable consumption were compared with a traditional didactic message. DESIGN: A Greco-Latin square, within-subjects experimental design was employed with 3 formats and 3 topics. SETTING: Community centers. PARTICIPANTS: Thirty-one ethnically diverse adults. INTERVENTION: Presented conversational, testimonial, and didactic nutrition messages. VARIABLES MEASURED: Believability, clarity, and perceived usefulness of messages presented; identification with sources in message; and self-efficacy with respect to produce consumption. ANALYSIS: Analysis of variance. RESULTS: Conversational formats were perceived as more believable than the control newsletter article. No differences were found with respect to clarity, usefulness, or self-efficacy. Identification with characters did significantly interact with message format in predicting the message assessment variables. However, the conversation and testimonial versions did not increase identification with persons in the message over the didactic presentation. CONCLUSIONS AND IMPLICATIONS: Audience response to narrative messages is contingent upon identification with persons portrayed in the message. The results underscore the need for careful pretesting of involvement with the story and identification with persons portrayed to maximize the effectiveness of narrative messages in health interventions. PMID- 14521826 TI - Nutrition education worksite intervention for university staff: application of the health belief model. AB - OBJECTIVE: To evaluate the efficacy of an 8-week worksite nutrition education intervention for university staff using the Health Belief Model (HBM) to promote healthful dietary behaviors that reduce risks for cardiovascular disease and cancer. DESIGN: 2 3 2 repeated measures baseline/posttest ex post facto research design. PARTICIPANTS: Staff employees were randomly assigned to treatment (n = 28) and control groups (n = 25). INTERVENTION: The intervention focused on specific health beliefs, nutrition knowledge, and dietary practices to demonstrate treatment effect. MAIN OUTCOME MEASURES: Dependent variables were specific health beliefs, nutrition knowledge, and dietary behaviors. Independent variables were demographic characteristics and group assignment. ANALYSES: Tests of parametric assumptions, power analyses, analysis of variance, and Kuder Richardson and Pearson product-moment coefficients were computed and specificity of treatment effects was assessed. RESULTS: Perceived benefits of healthy nutrition practices and nutrition knowledge related to cardiovascular disease and cancer significantly improved among the treatment participants, P <.001. Treatment group participants also significantly reduced total calories, fat, saturated fat, and cholesterol intake (each P <.001). CONCLUSIONS: The intervention appears to be related to treatment effects and significantly increased nutrition knowledge and decreased energy, fat, saturated fat, and cholesterol intake to levels consistent with national recommendations. PMID- 14521827 TI - Gem No. 363. Refrigerator art to promote 5 a day. PMID- 14521828 TI - Gem No. 364. Using food calendars to self-monitor: got 5? Nutrition for kids program. PMID- 14521829 TI - Gem No. 365. Fresh minds, from farm to classroom: a nutrition and agriculture game. PMID- 14521831 TI - A determinant for directionality of organelle transport in Drosophila embryos. AB - BACKGROUND: Motor-driven transport along microtubules is a primary cellular mechanism for moving and positioning organelles. Many cargoes move bidirectionally by using both minus and plus end-directed motors. How such cargoes undergo controlled net transport is unresolved. RESULTS: Using a combination of genetics, molecular biology, and biophysics, we have identified Halo, a novel regulator of lipid droplet transport in early Drosophila embryos. In embryos lacking Halo, net transport of lipid droplets, but not that of other cargoes, is specifically altered; net transport is minus-end directed at developmental stages when it is normally plus-end directed. This reversal is due to an altered balance of motion at the level of individual organelles; without Halo, travel distances and stall forces are reduced for plus-end and increased for minus-end motion. During development, halo mRNA is highly upregulated just as net plus-end transport is initiated (phase II), and its levels drop precipitously shortly before transport becomes minus-end directed (phase III). Exogenously provided Halo prevents the switch to net minus-end transport in phase III in wild type embryos and induces net plus-end transport during phase II in halo mutant embryos. This mechanism of regulation is likely to be of general importance because the Drosophila genome encodes a family of related proteins with similar sequences, each transiently expressed in distinct domains. CONCLUSIONS: We conclude that Halo acts as a directionality determinant for embryonic droplet transport and is the first member of a new class of transport regulators. PMID- 14521832 TI - Mathematical modeling suggests cooperative interactions between a disordered polyvalent ligand and a single receptor site. AB - BACKGROUND: The CDK inhibitor Sic1 must be phosphorylated on at least six sites in order to allow its recognition by the SCF ubiquitin ligase subunit Cdc4. However, because Cdc4 appears to have only a single phospho-epitope binding site, the apparent cooperative dependence on the number of phosphorylation sites in Sic1 cannot be accounted for by traditional thermodynamic models of cooperativity. RESULTS: We develop a general kinetic model, which predicts an unexpected multiplicative increase in affinity as a function of ligand sites. This effect, termed allovalency, derives from a high local concentration of interaction sites moving independently of each other. Modeling of this interaction by a first exit time approach indicates that the probability of ligand rebinding increases exponentially with the number of sites. This type of interaction is relatively immune to loss of any one site and may be easily tuned to any given threshold by adjusting the properties of individual sites. CONCLUSIONS: The allovalency model suggests that a previously undescribed mechanism may underlie certain cooperative interactions. The widespread occurrence of flexible polyvalent ligands in biological systems suggests that this principle may be broadly applicable. PMID- 14521833 TI - Intraflagellar transport is required in Drosophila to differentiate sensory cilia but not sperm. AB - BACKGROUND: Intraflagellar transport (IFT) uses kinesin II to carry a multiprotein particle to the tips of eukaryotic cilia and flagella and a nonaxonemal dynein to return it to the cell body. IFT particle proteins and motors are conserved in ciliated eukaryotes, and IFT-deficient mutants in algae, nematodes, and mammals fail to extend or maintain cilia and flagella, including sensory cilia. In Drosophila, the only ciliated cells are sensory neurons and sperm. no mechanoreceptor potential (nomp) mutations have been isolated that affect the differentiation and function of ciliated sense organs. The nompB gene is here shown to encode an IFT protein. Its mutant phenotypes reveal the consequences of an IFT defect in an insect. RESULTS: Mechanosensory and olfactory neurons in nompB mutants have missing or defective cilia. nompB encodes the Drosophila homolog of the IFT complex B protein IFT88/Polaris/OSM-5. nompB is expressed in the ciliated sensory neurons, and a functional, tagged NOMPB protein is located in sensory cilia and around basal bodies. Surprisingly, nompB mutant males produce normally elongated, motile sperm. Neuronally restricted expression and male germline mosaic experiments show that nompB-deficient sperm are fully functional in transfer, competition, and fertilization. CONCLUSIONS: NOMPB, the Drosophila homolog of IFT88, is required for the assembly of sensory cilia but not for the extension or function of the sperm flagellum. Assembly of this extremely long axoneme is therefore independent of IFT. PMID- 14521834 TI - Drosophila KAP interacts with the kinesin II motor subunit KLP64D to assemble chordotonal sensory cilia, but not sperm tails. AB - BACKGROUND: Kinesin II-mediated anterograde intraflagellar transport (IFT) is essential for the assembly and maintenance of flagella and cilia in various cell types. Kinesin associated protein (KAP) is identified as the non-motor accessory subunit of Kinesin II, but its role in the corresponding motor function is not understood. RESULTS: We show that mutations in the Drosophila KAP (DmKap) gene could eliminate the sensory cilia as well as the sound-evoked potentials of Johnston's organ (JO) neurons. Ultrastructure analysis of these mutants revealed that the ciliary axonemes are absent. Mutations in Klp64D, which codes for a Kinesin II motor subunit in Drosophila, show similar ciliary defects. All these defects are rescued by exclusive expression of DmKAP and KLP64D/KIF3A in the JO neurons of respective mutants. Furthermore, reduced copy number of the DmKap gene was found to enhance the defects of hypomorphic Klp64D alleles. Unexpectedly, however, both the DmKap and the Klp64D mutant adults produce vigorously motile sperm with normal axonemes. CONCLUSIONS: KAP plays an essential role in Kinesin II function, which is required for the axoneme growth and maintenance of the cilia in Drosophila type I sensory neurons. However, the flagellar assembly in Drosophila spermatids does not require Kinesin II and is independent of IFT. PMID- 14521835 TI - Targeting connexin43 expression accelerates the rate of wound repair. AB - The repair of tissue damage is a key survival process in all organisms and involves the coordinated activation of several cell types. Cell-cell communication is clearly fundamental to this process, and a great deal is known about extracellular communication within the wound site via cytokines. Here we show that direct cell-cell communication through connexin 43 (Cx43) gap junction channels also plays a major role in the wound healing process. In two different wound healing models, incisional and excisional skin lesions, we show that a single topical application of Cx43 antisense gel brings about a transient downregulation of Cx43 protein levels, and this results in a dramatic increase in the rate of wound closure. Cx43 knockdown reduces inflammation, seen both macroscopically, as a reduction in swelling, redness, and wound gape, and microscopically, as a significant decrease in neutrophil numbers in the tissue around the wound. One long-term consequence of the improved rate of healing is a significant reduction in the extent of granulation tissue deposition and the subsequent formation of a smaller, less distorted, scar. This approach is likely to have widespread therapeutic applications in other injured tissues and opens up new avenues of research into improving the wound healing process. PMID- 14521836 TI - Brief daily periods of binocular vision prevent deprivation-induced acuity loss. AB - The role of experience in the development of the central visual pathways has been explored in the past through examination of the consequences of imposed periods of continuously abnormal or biased visual input. The massive changes in the visual cortex (area 17) induced by selected early visual experience, especially monocular deprivation (MD) or experience (ME) where patterned visual input is provided to just one eye, are accompanied by profound and long-standing visual deficits. Although the use of exclusively abnormal experience permits identification of those aspects of the visual cortex and of visual function that can be influenced by visual experience during development, this approach may provide a distorted view of the nature of the role of visual experience because of the absence of any normal visual input. In this study a different approach was used whereby animals were provided daily with separate periods of normal (i.e., binocular exposure) and abnormal (monocular exposure) visual experience. We show that 2 hr of daily normal concordant binocular experience (BE) can outweigh or protect against much longer periods of monocular deprivation (MD) and permit the development of normal visual acuities in the two eyes. This result is not what would be expected if all visual input had equal influence on visual development. PMID- 14521837 TI - "Putting the face to the voice": matching identity across modality. AB - Speech perception provides compelling examples of a strong link between auditory and visual modalities. This link originates in the mechanics of speech production, which, in shaping the vocal tract, determine the movement of the face as well as the sound of the voice. In this paper, we present evidence that equivalent information about identity is available cross-modally from both the face and voice. Using a delayed matching to sample task, XAB, we show that people can match the video of an unfamiliar face, X, to an unfamiliar voice, A or B, and vice versa, but only when stimuli are moving and are played forward. The critical role of time-varying information is underlined by the ability to match faces to voices containing only the coarse spatial and temporal information provided by sine wave speech [5]. The effect of varying sentence content across modalities was small, showing that identity-specific information is not closely tied to particular utterances. We conclude that the physical constraints linking faces to voices result in bimodally available dynamic information, not only about what is being said, but also about who is saying it. PMID- 14521838 TI - Whole-genome analysis of 60 G protein-coupled receptors in Caenorhabditis elegans by gene knockout with RNAi. AB - G protein-coupled receptors (GPCRs) are the largest family of genes in animal genomes and represent more than 2% of genes in humans and C. elegans. These evolutionarily conserved seven-transmembrane proteins transduce a diverse range of signals. In view of their pivotal role in cell signaling, it is perhaps surprising that decades of genetic analysis in C. elegans, and recent genome-wide RNAi screens, have identified very few GPCR mutants. Therefore, we screened all GPCRs predicted to bind either small-molecule neurotransmitters or neuropeptides by using RNAi and quantitative behavioral assays. This shows that C16D6.2, C25G6.5, C26F1.6, F35G8.1, F41E7.3, and F59C12.2 are likely to be involved in reproduction, whereas C15B12.5, C10C6.2, C24A8.4, F15A8.5, F59D12.1, T02E9.1, and T05A1.1 have a role in locomotion. Gene deletions for F35G8.1 and T05A1.1 resulted in the same phenotype as that seen with RNAi. As some GPCRs may be resistant to RNAi, or may result in abnormalities not screened for here, the actual proportion of nonredundant receptors with an assayable function is probably greater. Strikingly, most phenotypes were observed for NPY-like receptors that may bind neuropeptides. This is consistent with the known actions of neuropeptides on the body wall muscle and reproductive tract in nematodes. PMID- 14521839 TI - Flt-1-dependent survival characterizes the epithelial-mesenchymal transition of colonic organoids. AB - Aberrant cell survival and resistance to apoptosis are hallmarks of tumor invasion and progression to metastatic disease, but the mechanisms involved are poorly understood. The epithelial-mesenchymal transition (EMT), a process that facilitates progression to invasive cancer, provides a superb model for studying such survival mechanisms. Here, we used a unique spheroid culture system that recapitulates the structure of the colonic epithelium and undergoes an EMT in response to cytokine stimulation to study this problem. Our data reveal that the EMT results in the increased expression of both VEGF and Flt-1, a tyrosine kinase VEGF receptor, and that the survival of these cells depends on a VEGF/Flt-1 autocrine pathway. Perturbation of Flt-1 function by either a blocking antibody or adenoviral expression of soluble Flt-1, which acts in a dominant-negative fashion, caused massive apoptosis only in cells that underwent EMT. This pathway was critical for the survival of other invasive colon carcinoma cell lines, and we observed a correlative upregulation of Flt-1 expression linked to in vivo human cancer progression. A role for Flt-1 in cell survival is unprecedented and has significant implications for Flt-1 function in tumor progression, as well as in other biological processes, including angiogenesis and development. PMID- 14521840 TI - Long-range communication between chromatin and microtubules in Xenopus egg extracts. AB - The mitotic spindle of animal cells is a bipolar array of microtubules that guides chromosome segregation during cell division. It has been proposed that during spindle assembly chromatin can positively influence microtubule stability at a distance from its surface throughout its neighboring cytoplasm. However, such an "a distance" effect has never been visualized directly. Here, we have used centrosomal microtubules and chromatin beads to probe the regulation of microtubule behavior around chromatin in Xenopus egg extracts. We show that, in this system, chromatin does affect microtubule formation at a distance, inducing preferential orientation of centrosomal microtubules in its direction. Moreover, this asymmetric distribution of microtubules is translated into a directional migration of centrosomal asters toward chromatin and their steady-state repositioning within 10 microm of chromatin. To our knowledge, this is the first direct evidence of a long-range guidance effect at the sub-cellular level. PMID- 14521841 TI - The Arabidopsis heterochronic gene ZIPPY is an ARGONAUTE family member. AB - Plants progress through a temporal sequence of juvenile, adult, and reproductive phases, each marked by the expression of phase-specific traits. Here we show that loss-of-function mutations in ZIPPY (ZIP) cause the premature expression of adult vegetative traits but do not accelerate the onset of reproductive competence or flowering time. ZIP encodes ARGONAUTE7 (AGO7), one of ten members of the ARGONAUTE family in Arabidopsis. In addition to playing developmental roles, some ARGONAUTE family members are required for RNAi-like phenomena, such as posttranscriptional gene silencing. In contrast to Arabidopsis ARGONAUTE1, ZIP has no significant role in transgene silencing; its primary function is in the regulation of developmental timing. PMID- 14521845 TI - Italy sets new hurdles for GM crops. PMID- 14521842 TI - Cell cycle-regulated transcription through the FHA domain of Fkh2p and the coactivator Ndd1p. AB - Recent studies in Saccharomyces cerevisiae by using global approaches have significantly enhanced our knowledge of the components involved in the transcriptional regulation of the cell cycle. The Mcm1p-Fkh2p complex, in combination with the coactivator Ndd1p, plays an important role in the cell cycle dependent expression of the CLB2 gene cluster during the G2 and M phases ([4-7]; see [8-10]for reviews). Fkh2p is phosphorylated in a cell cycle-dependent manner, and peak phosphorylation occurs coincidentally with maximal expression of Mcm1p Fkh2p-dependent gene expression. However, the mechanism by which this complex is activated in a cell cycle-dependent manner is unknown. Here, we demonstrate that the forkhead-associated (FHA) domain of Fkh2p directs cell cycle-regulated transcription and that the activity of this domain is dependent on the coactivator Ndd1p. Ndd1p was found to be phosphorylated in a cell cycle-dependent manner by Cdc28p-Clb2p, and, importantly, this phosphorylation event promotes interactions between Ndd1p and the FHA domain of Fkh2p. Furthermore, mutation of the FHA domain blocks these phosphorylation-dependent interactions and abolishes transcriptional activity. Our data therefore link the transcriptional activity of the FHA domain with cell cycle-dependent phosphorylation of the coactivator Ndd1p and reveal a mechanism that permits precise temporal activation of the Mcm1p Fkh2p complex. PMID- 14521847 TI - Passion flowers. PMID- 14521846 TI - 50 years before the double helix. PMID- 14521848 TI - Leeches. PMID- 14521849 TI - Medicine man. PMID- 14521850 TI - Listeria monocytogenes rotates around its long axis during actin-based motility. PMID- 14521851 TI - Cell migration. PMID- 14521852 TI - John Gurdon. PMID- 14521853 TI - Gravity perception: how plants stand up for themselves. PMID- 14521854 TI - Spliceosomal introns: new insights into their evolution. AB - A new genome-wide analysis of spliceosomal introns indicates massive loss and gain of introns has taken place in many eukaryotic lineages. Only a small subset of the analyzed introns was present in the common ancestor of plants, fungi, animals and Plasmodium. PMID- 14521855 TI - Auditory physiology: listening with K+ channels. PMID- 14521856 TI - Algal morphogenesis: how volvox turns itself inside-out. PMID- 14521857 TI - EGF receptor signaling: a prickly proposition. PMID- 14521858 TI - Phototransduction: shedding light on translocation. AB - Light induces the migration of arrestin to the photosensitive membrane in both vertebrate and invertebrate photoreceptors. New work has identified a phosphoinositide lipid binding domain in Drosophila arrestin and implicates PIP(3) in control of arrestin translocation. PMID- 14521859 TI - Brain mapping: new wave optical imaging. AB - Optical imaging of intrinsic signals is widely used for high-resolution brain mapping in various animal species. A new approach using continuous data acquisition and Fourier decomposition of the signal allows for much faster mapping, opening up the possibility of applying this method to new experimental questions. PMID- 14521860 TI - Plant cell polarity: the ins-and-outs of sterol transport. PMID- 14521861 TI - Acetylcholine: cognitive and brain functions. PMID- 14521862 TI - Muscarinic cholinergic influences in memory consolidation. AB - The central cholinergic system and muscarinic cholinergic receptor (mR) activation have long been associated with cognitive function. Although mR activation is no doubt involved in many aspects of cognitive functioning, the extensive evidence that memory is influenced by cholinergic treatments given after training either systemically or intra-cranially clearly indicates that cholinergic activation via mRs is a critical component in modulation of memory consolidation. Furthermore, the evidence indicates that activation of mRs in the basolateral amygdala (BLA) plays an essential role in enabling other neuromodulatory influences on memory consolidation. Memory can also be affected by posttraining activation of mRs in the hippocampus, striatum and cortex. Evidence of increases in hippocampal and cortical acetylcholine (ACh) levels following learning experiences support the view that endogenous ACh release is involved in long-term memory consolidation. Furthermore, the findings indicating that mR drug treatments influence plasticity in the hippocampus and in sensory cortices strongly suggest that mR activation is involved in the storage of information in these brain regions. PMID- 14521863 TI - Acetylcholine modulation of neural systems involved in learning and memory. AB - Extensive evidence supports the view that cholinergic mechanisms modulate learning and memory formation. This paper reviews evidence for cholinergic regulation of multiple memory systems, noting that manipulations of cholinergic functions in many neural systems can enhance or impair memory for tasks generally associated with those neural systems. While parallel memory systems can be identified by combining lesions with carefully crafted tasks, most-if not all tasks require the combinatorial participation of multiple neural systems. This paper offers the hypothesis that the magnitude of acetylcholine (ACh) release in different neural systems may regulate the relative contributions of these systems to learning. Recent studies of ACh release, obtained with in vivo microdialysis samples during training, together with direct injections of cholinergic drugs into different neural systems, provide evidence that release of ACh is important in engaging these systems during learning, and that the extent to which the systems are engaged is associated with individual differences in learning and memory. PMID- 14521864 TI - Role of cholinergic system on the construction of memories: taste memory encoding. AB - There is a large body of evidence suggesting that cholinergic activity is involved in memory processes. It seems that cholinergic activity is essential to learn several tasks and recent works suggest that acetylcholine plays an important role during the early stages of memory formation. In this review, we will discuss the results related to taste memory formation, focusing particularly on the conditioned taste aversion paradigm. We will first give evidence that nucleus basalis magnocellularis is involved in taste memory formation, due to its cholinergic projections. We then show that the cholinergic activity of the insular (gustatory) cortex is related to the taste novelty, and that the cholinergic signals initiated by novelty are crucial for taste memory formation. Then we present recent data indicating that cortical activation of muscarinic receptors is necessary for taste trace encoding, and also for its consolidation under certain circumstances. Finally, interactions between the cholinergic and other neuromodulatory systems inducing intracellular mechanisms related to plastic changes will be proposed as important processes underlying gustatory memory trace storage. PMID- 14521865 TI - Modulation of cholinergic transmission enhances excitability of hippocampal pyramidal neurons and ameliorates learning impairments in aging animals. AB - Four cholinesterase inhibitors have been approved by the US Food and Drug Administration for treating behavioral symptoms of Alzheimer's disease. Here we review our experiences with two cholinesterase inhibitors (metrifonate and galanthamine) and a muscarinic acetylcholine receptor agonist (CI-1017) in behavioral pharmacological and brain slice experiments in aging and young rabbits. Aging rabbits are impaired in their ability to acquire the hippocampus dependent trace eyeblink conditioning task, as compared to young controls. A large proportion of aging animals cannot reach behavioral criterion in this task. Those that do learn, do so more slowly. In addition, the post-burst afterhyperpolarization and spike frequency accommodation is increased in hippocampal pyramidal neurons from aging animals, i.e., cellular excitability is reduced as compared to those from young animals. Metrifonate, galanthamine, and CI-1017 reduced the learning deficits observed in aging rabbits so that they learned almost as quickly as young controls. These cholinergic compounds also enhanced the postsynaptic excitability of hippocampal pyramidal neurons in vitro. Therefore, we propose that the amelioration of learning impairment with the cholinergic compounds may in part be due to the enhanced excitability of hippocampal pyramidal neurons. The potential relevance of our studies to further understanding the cellular and behavioral changes that occur with normal aging and Alzheimer's Disease is discussed. PMID- 14521867 TI - Attentional functions of cortical cholinergic inputs: what does it mean for learning and memory? AB - The hypothesis that cortical cholinergic inputs mediate attentional functions and capacities has been extensively substantiated by experiments assessing the attentional effects of specific cholinotoxic lesions of cortical cholinergic inputs, attentional performance-associated cortical acetylcholine release, and the effects of pharmacological manipulations of the excitability of basal forebrain corticopetal cholinergic projections on attentional performance. At the same time, numerous animal experiments have suggested that the integrity of cortical cholinergic inputs is not necessary for learning and memory, and a dissociation between the role of the cortical cholinergic input system in attentional functions and in learning and memory has been proposed. We speculate that this dissociation is due, at least in part, to the use of standard animal behavioral tests for the assessment of learning and memory which do not sufficiently tax defined attentional functions. Attentional processes and the allocation of attentional capacities would be expected to influence the efficacy of the acquisition and recall of declarative information and therefore, persistent abnormalities in the regulation of the cortical cholinergic input system may yield escalating impairments in learning and memory. Furthermore, the cognitive effects of loss of cortical cholinergic inputs are augmented by the disruption of the top-down regulation of attentional functions that normally acts to optimize information processing in posterior cortical areas. Because cortical cholinergic inputs play an integral role in the mediation of attentional processing, the activity of cortical cholinergic inputs is hypothesized to also determine the efficacy of learning and memory. PMID- 14521866 TI - Cholinergic modulation of learning and memory in the human brain as detected with functional neuroimaging. AB - The advent of neuroimaging methods such as functional magnetic resonance imaging (fMRI) and positron emission tomography (PET) has provided investigators with a tool to study neuronal processes involved in cognitive functions in humans. Recent years have seen an increasing amount of studies which mapped higher cognitive functions to specific brain regions. These studies have had a great impact on our understanding of neuroanatomical correlates of learning and memory in the living human brain. Recently, advances were made to go beyond the use of fMRI as a pure cognitive brain mapping device. One of these advances includes the use of psychopharmacological approaches in conjunction with neuroimaging. The paper will introduce the combination of neuroimaging and psychopharmacology as a tool to study neurochemical modulation of human brain function. A review of imaging studies using cholinergic challenges in the context of explicit and implicit learning and memory paradigms is provided which show that cholinergic neurotransmission modulates task-related activity in sensory and frontal cortical brain areas. PMID- 14521868 TI - Acetylcholine actions in the dorsomedial striatum support the flexible shifting of response patterns. AB - There is accumulating evidence that the dorsomedial striatum plays a significant role in the learning of a new response pattern and the inhibiting of old response patterns when conditions demand a shift in strategies. This paper proposes that activity of cholinergic neurons in the dorsomedial striatum is critical for enabling behavioral flexibility when there is a change in task contingencies. Recent experimental findings are provided supporting this idea. Measuring acetylcholine efflux from the dorsomedial striatum during the acquisition and reversal learning of a spatial discrimination shows that acetylcholine efflux selectively increases during reversal learning as a rat begins to learn a newly reinforced spatial location, but returns to near basal levels when a rat reliably executes the new choice pattern. Experimental findings are also described indicating that the blockade of muscarinic cholinergic receptors in the dorsomedial striatum does not impair acquisition of an egocentric response discrimination, but impairs reversal learning of an egocentric response discrimination. Based on these results, increased cholinergic activity at muscarinic receptors is part of a neurochemical process in the dorsomedial striatum that allows inhibition of a previously relevant response pattern while learning a new response pattern. In situations that demand behavioral flexibility, muscarinic cholinergic activity in the dorsomedial striatum may directly influence corticostriatal plasticity to produce changes in response patterns. PMID- 14521869 TI - The nucleus basalis and memory codes: auditory cortical plasticity and the induction of specific, associative behavioral memory. AB - Receptive field (RF) plasticity develops in the primary auditory cortex (ACx) when a tone conditioned stimulus (CS) becomes associated with an appetitive or aversive unconditioned stimulus (US). This prototypical stimulus-stimulus (S-S) association is accompanied by shifts of frequency tuning of neurons toward or to the frequency of the CS such that the area of best tuning of the CS frequency is increased in the tonotopic representation of the ACx. RF plasticity has all of the major characteristics of behavioral associative memory: it is highly specific, discriminative, rapidly induced, consolidates (becomes stronger and more specific over hours to days) and can be retained indefinitely (tested to two months). Substitution of nucleus basalis (NB) stimulation for a US induces the same associative RF plasticity, and this requires the engagement of muscarinic receptors in the ACx. Pairing a tone with NB stimulation actually induces specific, associative behavioral memory, as indexed by post-training frequency generalization gradients. The degree of acquired behavioral significance of sounds appears to be encoded by the number of neurons that become retuned in the ACx to that acoustic stimulus, the greater the importance, the greater the number of re-tuned cells. This memory code has recently been supported by direct neurobehavioral tests. In toto, these findings support the view that specific, learned auditory memory content is stored in the ACx, and further that this storage of information during learning and the instantiation of the memory code involves the engagement of the nucleus basalis and its release of acetylcholine into target structures, particularly the cerebral cortex. PMID- 14521870 TI - Regulation of glutamate synapses by nicotinic acetylcholine receptors in auditory cortex. AB - Acetylcholine plays an important role in regulating the processing of sensory stimuli, and understanding its specific cellular actions is critical to understanding how sensory cortex develops and functions in different behavioral states. Here we review recent work on the cellular effects of nicotinic receptor activation in auditory cortex and describe how these actions could affect systems level auditory function. In particular, we describe a novel function of nicotinic acetylcholine receptors to regulate glutamate synapses containing N-methyl-D aspartate receptors during early postnatal development. The transient regulation of developing glutamate synapses also defines a window of vulnerability during which exposure to exogenous nicotine disrupts synapse development. Thus, it appears that nicotinic regulation of glutamate synapses is a critical feature of auditory cortex development. PMID- 14521871 TI - Contribution of acetylcholine to visual cortex plasticity. AB - Acetylcholine is involved in a variety of brain functions. In the visual cortex, the pattern of cholinergic innervation varies considerably across different mammalian species and across different cortical layers within the same species. The physiological effects of acetylcholine in the visual cortex display complex responses, which are likely due to cholinergic receptor subtype composition in cytoplasm membrane as well as interaction with other transmitter systems within the local neural circuitry. The functional role of acetylcholine in visual cortex is believed to improve the signal-to-noise ratio of cortical neurons during visual information processing. Available evidence suggests that acetylcholine is also involved in experience-dependent visual cortex plasticity. At the level of synaptic transmission, activation of muscarinic receptors has been shown to play a permissive role in visual cortex plasticity. Among the muscarinic receptor subtypes, the M(1) receptor seems to make a predominant contribution towards modifications of neural circuitry. The signal transduction cascade of the cholinergic pathway may act synergistically with that of the NMDA receptor pathway, whose activation is a prerequisite for cortical plasticity. PMID- 14521872 TI - Enhanced cholinergic suppression of previously strengthened synapses enables the formation of self-organized representations in olfactory cortex. AB - Computational modeling assists in analyzing the specific functional role of the cellular effects of acetylcholine within cortical structures. In particular, acetylcholine may regulate the dynamics of encoding and retrieval of information by regulating the magnitude of synaptic transmission at excitatory recurrent connections. Many abstract models of associative memory function ignore the influence of changes in synaptic strength during the storage process and apply the effect of these changes only during a so-called recall-phase. Efforts to ensure stable activity with more realistic, continuous updating of the synaptic strength during the storage process have shown that the memory capacity of a realistic cortical network can be greatly enhanced if cholinergic modulation blocks transmission at synaptic connections of the association fibers during the learning process. We here present experimental data from an olfactory cortex brain slice preparation showing that previously potentiated fibers show significantly greater suppression (presynaptic inhibition) by the cholinergic agonist carbachol than unpotentiated fibers. We conclude that low suppression of non-potentiated fibers during the learning process ensures the formation of self organized representations in the neural network while the higher suppression of previously potentiated fibers minimizes interference between overlapping patterns. We show in a computational model of olfactory cortex, that, together, these two phenomena reduce the overlap between patterns that are stored within the same neural network structure. These results further demonstrate the contribution of acetylcholine to mechanisms of cortical plasticity. The results are consistent with the extensive evidence supporting a role for acetylcholine in encoding of new memories and enhancement of response to salient sensory stimuli. PMID- 14521873 TI - Short-term estrogen treatment in ovariectomized rats augments hippocampal acetylcholine release during place learning. AB - Estrogen modulates learning and memory in ovariectomized and naturally cycling female rats, especially in tasks using spatial learning and navigation. Estrogen also modulates cholinergic function in various forebrain structures. Past studies have shown positive correlations between hippocampal ACh output and performance on hippocampus-dependent tasks. The present study examined whether estradiol replacement would potentiate hippocampal ACh release during place learning. In vivo microdialysis and HPLC were used to measure extracellular ACh levels in the hippocampus of ovariectomized female rats that had received s.c. injections of 17beta-estradiol (10 microg) or sesame oil (vehicle treatment) 48 and 24h prior to training on a place task. Estrogen did not alter baseline levels of extracellular ACh in the hippocampus. During training, hippocampal ACh increased in ovariectomized rats regardless of estrogen status. However, while estradiol did not enhance learning in this experiment, estradiol significantly potentiated the increase in hippocampal ACh release seen during place training. This represents the first demonstration of on-line assessment of ACh output in hippocampus during learning in female rats and suggests that estrogen-dependent modulation of ACh release during training might control activation of different neural systems used during learning. PMID- 14521874 TI - Cholinergic dependence of taste memory formation: evidence of two distinct processes. AB - Learning the aversive or positive consequences associated with novel taste solutions has a strong significance for an animal's survival. A lack of recognition of a taste's consequences could prevent ingestion of potential edibles or encounter death. We used conditioned taste aversion (CTA) and attenuation of neophobia (AN) to study aversive and safe taste memory formation. To determine if muscarinic receptors in the insular cortex participate differentially in both tasks, we infused the muscarinic antagonists scopolamine at distinct times before or after the presentation of a strong concentration of saccharin, followed by either an i.p. injection of a malaise-inducing agent or no injection. Our results showed that blockade of muscarinic receptors before taste presentation disrupts both learning tasks. However, the same treatment after the taste prevents AN but not CTA. These results clearly demonstrate that cortical cholinergic activity participates in the acquisition of both safe and aversive memory formation, and that cortical muscarinic receptors seem to be necessary for safe but not for aversive taste memory consolidation. These results suggest that the taste memory trace is processed in the insular cortex simultaneously by at least two independent mechanisms, and that their interaction would determine the degree of aversion or preference learned to a novel taste. PMID- 14521875 TI - Cholinergic modulation of the hippocampus during encoding and retrieval. AB - The present experiments were aimed at determining whether acetylcholine (ACh) plays a role in encoding and retrieval of spatial information using a modified Hebb-Williams maze. In addition, the present experiments tested two computational models of hippocampal function during encoding and retrieval using a maze sensitive to hippocampal disruption. Thirty male, Long-Evans rats served as subjects. Chronic cannulae were implanted bilaterally into the CA3 (n=26) and CA1 (n=5) subregions of the hippocampus. Rats were tested using a modified Hebb Williams maze. In the first experiment, rats were injected with either saline or scopolamine hydrobromide 10 min before testing for each day. The number of errors made per day per group was used as the measure of learning. Encoding was assessed by the average number of errors made on the first five trials of Day 1 compared to the last five trials of Day 1, whereas the average number of errors made on the first five trials of Day 2 compared to the last five trials of Day I was used to assess retrieval. No deficit was found for the saline group. The scopolamine group showed a deficit in encoding, but not retrieval. In the second experiment, rats were injected with either saline or physostigmine 10 min before testing each day. In contrast to the scopolamine groups, the physostigmine group showed a deficit in retrieval, but not encoding. To test whether the retrieval deficit was due to a disruption in storage or gaining access to the information two groups of rats received either saline on Day 1 and physostigmine on Day 2 or physostigmine on Day 1 and saline on Day 2. In addition, one group received physostigmine immediately after testing on Day 1. Data indicate that physostigmine causes a disruption of retrieval by means of a disruption in consolidation process. In conclusion, the cholinergic antagonist, scopolamine, disrupts encoding in both CA3 and CA1 subregions of the hippocampus. Furthermore, the cholinesterase inhibitor, physostigmine, boosts ACh action during a time when cholinergic levels need to decline for proper consolidation. PMID- 14521876 TI - Translocation model of Candida albicans in DBA-2/J mice with protein calorie malnutrition mimics hematogenous candidiasis in humans. AB - To elucidate the mechanism of translocation of Candida albicans from the intestine to the bloodstream, we attempted to establish a murine model for hematogenous translocation of C. albicans using DBA-2/J mice with protein calorie malnutrition (PCM). PCM severely affected the development of the intestinal epithelia; thereby, the keratin and mucinous layers became very thin. Oral inoculation with C. albicans resulted in long-term colonization in the intestine of the PCM mice but not the well-nourished animals. Chemotherapy with a combination of cyclophosphamide and methotrexate, which started four days after oral inoculation of C. albicans, resulted in the systemic dissemination of C. albicans from the intestine in the PCM mice. Among systemic organs, C. albicans was first isolated from the liver, in which focal necrosis, containing fungal balls of yeast-like forms and/or hyphae, was formed. Subsequently, C. albicans was first recovered from the blood of the infected PCM mice at one day after the isolation from the liver, and thereafter, candidemia continued to increase its intensity until death. Histological study indicated that C. albicans gained entry into the systemic organs from the epithelia of the esophago-cardiac junction as well as the Ileo-cecal portions of the infected mice. The results of our present study therefore suggest that this PCM mouse model may be useful for better understanding of the chemotherapy-induced translocation by C. albicans from the gut to the systemic organs in compromised humans. PMID- 14521877 TI - Studies of aerolysin promoters from different Aeromonas spp. AB - Aeromonas spp. have been related to food and waterborne diseases. The pore forming toxin aerolysin is regarded as the most important virulence factor in Aeromonas food poisoning. In this work the aerolysin promoters from several Aeromonas spp. strains have been sequenced, and divided into two sequence groups. Further analyses of the promoters were carried out in a reporter-plasmid, pSTINA II. This plasmid was constructed as a hybrid of pUC4K, pACYC184 and pKK232-8. We could conclude that our constructed reporter-gene plasmid was functional and was used to compare different promoters in an aerolysin negative Aeromonas spp. This construct made it possible to study the expression of the reporter gene using different aerolysin promoters under several conditions. We were able to show that the two obtained sequence groups of aerolysin promoters gave different expression of the reporter gene, and that this expression was dependent of temperature and osmolarity. Reducing the size of one promoter sequence from 254 to 148 bp and 102 bp gave a gradual reduction of the reporter gene expression under all conditions. According to our assay there seems to be more than one functional promoter upstream of the aerolysin gene, although the RT-PCR indicated one transcription starting point, under all test conditions. PMID- 14521878 TI - Occurrence of the Yersinia high-pathogenicity island and iron uptake systems in clinical isolates of Klebsiella pneumoniae. AB - The ability to acquire iron is crucial to bacteria during an infection. Thirty four strains of Klebsiella pneumoniae isolated from clinical specimens were examined for the use of various strategies to obtain iron. The isolates employed several iron uptake mechanisms, including production of enterobactin (100%) and aerobactin (50%). Few isolates (18%) produced yersiniabactin, a siderophore encoded by the Yersinia high-pathogenicity island (HPI) despite genetic diversity of the HPI. Majority of the isolates used human transferrin (74%), lactoferrin (97%), hemoglobin (74%), and hemoglobin-haptoglobin complex (56%) as a sole source of iron. Multiple iron uptake systems may be of benefit to the bacteria during infection. PMID- 14521879 TI - Characterization of a bovine lactoferrin binding protein of Streptococcus uberis. AB - The interaction between Streptococcus uberis and bovine lactoferrin (bLf) has been characterized. The binding of 125I-bLf to S. uberis was time-dependent and displaceable by unlabeled bLf. The Scatchard plot was linear and approximately 7,800 binding sites were expressed by each bacterial cell, with an affinity of 1.0 x 10(-7) M. Both heat and protease treatment of bacterial cells reduced bLf binding significantly, indicating the presence of a cell surface localized protein receptor for the glycoprotein. One protein was identified from the cell wall of S. uberis as the functionally active bLf-binding protein and it existed in both monomeric and dimeric forms. The recombinant protein expressed in E. coli cells was able to bind bLf and had molecular weights similar to that of native S. uberis. Deletion analysis located the bLf-binding domain to a 200 amino acid region at the amino terminus of Lbp. Analysis of the primary and secondary structure suggested that Lbp is an M-like protein. An isogenic mutant of S. uberis lacking the internal sequence of the lbp gene was constructed by allele replacement. Adherence experiments with wild type S. uberis and the lbp mutant revealed that Lbp is not responsible for attachment of S. uberis to host epithelial cells. PMID- 14521880 TI - Reduced virulence of a hfq mutant of Pseudomonas aeruginosa O1. AB - The Sm-like protein Hfq has been implicated in the regulation of sigmaS-dependent and sigmaS-independent genes in E. coli and in the regulation of virulence factors in both, Yersinia enterocolitica and Brucella abortus. Here, we have studied the effect of Hfq on virulence and stress response of Pseudomonas aeruginosa (PAO1). We have constructed a PAO1hfq- mutant and a PAO1hfq-rpoS- double mutant to permit distinction between direct and indirect effects of Hfq. When compared to the wild-type and the rpoS- strains, the hfq knock out strain showed a reduced growth rate and was unable to utilize glucose as a sole carbon source. Elastase activity was 80% reduced in the hfq- mutant when compared to the wild-type or the rpoS- strain, whereas alginate production seemed to be solely affected by sigmaS. The production of catalase and pyocyanin was shown to be affected in an additive manner by both, Hfq and sigmaS. Moreover, twitching and swarming mediated by typeIV pili was shown to be impaired in the hfq- mutant. When compared to PAO1 wild-type and the rpoS- mutant, the hfq- mutant decreased virulence in Galleria mellonella by a factor of 1 x 10(4) and 5 x 10(3), respectively. Likewise, when compared to wild-type, the PAO1hfq- mutant was significantly attenuated in virulence when administered intraperitoneally in mice. These results strongly suggest that Hfq is a global regulator of PAO1 virulence and stress response which is not exclusively due to its role in stimulating the synthesis of sigmaS. PMID- 14521881 TI - Roles of conserved nucleotide-binding domains in accessory proteins, HypB and UreG, in the maturation of nickel-enzymes required for efficient Helicobacter pylori colonization. AB - Helicobacter pylori synthesizes two nickel-containing enzymes (urease and hydrogenase), both of which are important pathogenesis factors. Among the many accessory proteins needed for maturation of these Ni-enzymes, are two proteins, HypB and UreG, each of which contain a conserved nucleotide-binding domain (GSGKT). To address the role of this domain in the maturation process, site directed mutations were introduced in both hypB and ureG. The hypB site-directed mutant strain (Lys59 to Ala59) lacked hydrogenase activity and had less than 1% of the parental urease activity. Hydrogenase activity was partially, and urease activity was fully restored in the hypB mutant strain when grown on nickel supplemented media. The hydrogenase activity of the ureG site-directed mutant strain (Lys14 to Ala14) was comparable to that of the parental strain. However, the ureG mutant strain lacked urease activity, and this deficiency could not be suppressed even when the strain was grown on nickel supplemented media. The expression of immunologically detectable HypB and UreG in the mutants was similar to the parental strain. Expression of the UreA and UreB subunits of urease in both the mutants was also normal. Purified UreG parental and mutant (Lys14 to Ala14) proteins had molecular masses of 27 kDa, but possessed negligible GTP hydrolyzing activity. PMID- 14521882 TI - MRI appearances of mullerian duct abnormalities. AB - Mullerian duct abnormalities are a well-documented cause of sub-fertility, and in some cases can be successfully treated in order to preserve fertility and prevent complications. In this paper we review the various types of anomaly and present their appearances as demonstrated by magnetic resonance imaging (MRI). MRI provides an excellent, non-invasive method of differentiating between the different types of anomaly and defining their suitability for surgical treatment. PMID- 14521883 TI - MRI features of tuberculosis of peripheral joints. AB - The aim of this article is to present the magnetic resonance imaging (MRI) features of peripheral tubercular arthritis. The clinical presentation of peripheral tubercular arthritis is variable and simulates other chronic inflammatory arthritic disorders. MRI is a highly sensitive technique which demonstrates fine anatomical details and identifies the early changes of arthritis, which are not visible on radiographs. The MRI features of tubercular arthritis include synovitis, effusion, central and peripheral erosions, active and chronic pannus, abscess, bone chips and hypo-intense synovium. These imaging features in an appropriate clinical setting may help in the diagnosis of tubercular arthritis. Early diagnosis and treatment can effectively eliminate the long-term morbidity of joints affected by tuberculosis. PMID- 14521884 TI - MRI of Paget's disease of bone. AB - Paget's disease is a relatively common condition in the elderly population and is not uncommonly seen as an incidental finding on imaging studies, including magnetic resonance imaging (MRI). It is important, therefore, for the radiologist to be aware of the MRI manifestations of the condition, so as not to mistake it for more sinister pathology. MRI can also be used to investigate the various complications of Paget's disease, particularly sarcomatous degeneration. This review describes the appearances of uncomplicated Paget's disease during its different pathological stages and in the setting of its various complications. PMID- 14521885 TI - Doppler ultrasound for detection of renal transplant artery stenosis-threshold peak systolic velocity needs to be higher in a low-risk or surveillance population. PMID- 14521886 TI - Doppler ultrasound for detection of renal transplant artery stenosis-threshold peak systolic velocity needs to be higher in a low-risk or surveillance population. AB - AIMS: To establish the ideal threshold arterial velocity for the diagnosis of renal transplant artery stenosis in a surveillance population with a low pre-test probability of stenosis. METHODS: Retrospective review of Doppler ultrasound, angiographic and clinical outcome data of patients transplanted over a 3-year period. Data used to calculate sensitivity, specificity, positive predictive values (PPV) and negative predictive values (NPV) for various threshold peak systolic velocity values. RESULTS: Of 144 patients transplanted, full data were available in 117 cases. Five cases had renal transplant artery stenosis-incidence 4.2% [stenosis identified at a mean of 6.5 months (range 2-10 months)]. All five cases had a significant arterial pressure gradient across the narrowing and underwent angioplasty. Threshold peak systolic velocity of > or =2.5 m/s is not ideal [specificity=79% (CI 65-82%), PPV=18% (CI 6-32%), NPV=100% (CI 94-100%)], subjecting many patients to unnecessary angiography-8/117 (6%) in our population. Comparable values if the threshold is set at > or =3.0 m/s are 93% (CI 77-96%), 33% (CI 7-44%) and 99% (CI 93-100%), respectively. The clinical outcome of all patients was satisfactory, with no unexplained graft failures or loss. CONCLUSIONS: In a surveillance population with a low pre-test probability of stenosis, absolute renal artery velocity > or =2.5 m/s is a limited surrogate marker for significant renal artery stenosis. The false-positive rate is high, and > or =3.0 m/s is a better choice which will halve the number of patients enduring unnecessary angiography. Close clinical follow-up of patients in the 2.5 3.0 m/s range, with repeat Doppler ultrasound if necessary, will identify the test false-negatives. PMID- 14521887 TI - Early-enhancing non-neoplastic lesions on gadolinium-enhanced MRI of the liver. AB - AIM: To assess the frequency, cause, and significance of early-enhancing, non neoplastic (EN) lesions on gadolinium-enhanced magnetic resonance imaging (MRI) of the liver performed for the detection of malignant hepatic tumours. MATERIALS AND METHODS: From September 1997 to September 2000, we reviewed the images of 125 patients, suspected of having hepatic tumours, in whom (1) gadolinium-enhanced triphasic dynamic gradient-recalled-echo (GRE) imaging in addition to unenhanced T1- and T2-weighted MRI was performed, (2) conventional angiography and combination computed tomography (CT) hepatic arteriography and CT during arterial portography were performed within 2 weeks of the MRI, and (3) definitive surgery within 2 weeks of the MRI or follow-up study by means of intravenously contrast enhanced CT or MRI in 10 months or more was performed. Angiographic studies were correlated to determine the underlying causes of the EN lesions. RESULTS: We found 78 EN lesions in 36 patients (29%), ranging in size from 4 and 50 mm (mean, 12.2 mm). From the MR reports, our radiologists had prospectively diagnosed EN lesions as probable malignant tumours in eight (10%), possible malignant tumours in 36 (46%), and probable non-neoplastic lesion in 34 (44%). EN lesions were found in 27 of 81 (33%) cirrhotic patients and in nine of 44 (20%) non-cirrhotic patients. Fifty-one EN lesions (65%) were located along the liver edge. The shape was circular in 42 (54%), oval in 14 (18%), irregular in 12 (15%), wedge-shaped in seven (9%), and fan-shaped in three (4%). Twenty EN lesions (26%) appeared slightly hyperintense on T2-weighted images. The causes were non-neoplastic arterio-portal shunting in 48 (62%), cystic venous drainage in four (5%), rib compression in four (5%), aberrant right gastric venous drainage in two (3%), and unknown in 20 (26%). CONCLUSION: Over half the number of EN lesions were caused by non-neoplastic arterio-portal shunting, occasionally showing slight hyperintensity on T2-weighted images. On MR images the non-neoplastic nature of the EN lesion was often ascertained. Radiologists should not overcall EN lesions as malignant as the patients involved would be inappropriately considered inoperable. In problematic cases, further investigation with angiographic CT or follow-up imaging studies should be performed. PMID- 14521888 TI - Technetium-99m tetrofosmin single photon emission computed tomography to detect metastatic papillary thyroid carcinoma in patients with elevated human serum thyroglobulin levels but negative I-131 whole body scan. AB - AIM: The aim of this study was to evaluate the effectiveness of technetium-99m tetrofosmin (Tc-99m TF) single photon emission computed tomography (SPECT) of the neck and chest to detect metastatic lesions in papillary thyroid carcinoma (PTC) after near total thyroidectomy and radioiodine (I-131) treatment in patients who present with elevated serum human thyroglobulin (hTg) levels but negative I-131 whole body scan (WBS). MATERIALS AND METHODS: Twenty patients with PTC treated by near total thyroidectomy and I-131 treatments were included in this study. All 20 patients had negative I-131 WBS results and elevated hTg levels (hTg 2.0 microIU/ml) under thyroid-stimulating hormone (TSH) stimulation (TSH 30 microIU/ml). Nineteen of the 20 cases were confirmed to have metastases by operation/biopsy histopathological findings or clinical follow-up longer than 1 year by additional morphological imaging techniques. The remaining patient has been followed up closely and has been disease free for 10 months. Tc-99m TF SPECT was performed to detect metastatic lesions. RESULTS: Tc-99m TF SPECT demonstrated lesions in 11/19 patients; a sensitivity of 57.9%. Tc-99m TF SPECT failed to demonstrate lesions in eight patients including smaller lymph nodes and miliary lung metastases. CONCLUSIONS: We conclude that Tc-99m TF SPECT is a useful additional tool to detect metastatic lesions in PTC with elevated hTg but negative I-131 WBS. However, smaller lymph nodes and miliary lung metastases may be missed. PMID- 14521889 TI - CT-guided lung biopsy: factors influencing diagnostic yield and complication rate. AB - AIM: To determine factors influencing diagnostic yield in computed tomography (CT)-guided biopsy of lung lesions. MATERIALS AND METHODS: One hundred and ninety five consecutive CT-guided lung biopsies were performed in 182 patients between August 1995 and September 2000 and either fine-needle aspirate samples for cytology or core biopsy samples for histology were collected. Procedures were divided into a diagnostic group (true-positive and true-negative results) and a non-diagnostic group (false-positive and false-negative results) and the factors affecting diagnostic accuracy assessed. RESULTS: One hundred and fifty-six lesions (86%) were malignant, and 26 (14%) were benign. More than one biopsy was performed for 12 lesions. One hundred and thirty-two biopsies were true-positive, 27 true-negative and 36 false-negative. No false-positive results occurred in the study. Overall diagnostic accuracy was 81.5%. Significantly more core biopsies than fine-needle aspirates were diagnostic: 93 versus 78% (p<0.005). No difference was found in frequency of pneumothorax between these two groups. There was a difference in the average depth from the pleural surface of lesions in the diagnostic and non-diagnostic groups, but this did not attain statistical significance: 9.8 versus 17.2 mm (p=0.054). CONCLUSION: In this study CT-guided lung biopsy core biopsy was a more accurate method of tissue sampling than fine needle aspiration, and was not associated with an excess of complications. PMID- 14521890 TI - Scaphoid and pronator fat stripes are unreliable soft tissue signs in the detection of radiographically occult fractures. AB - AIM: To determine the clinical value of scaphoid and pronator fat stripes in identifying occult underlying scaphoid and distal radius fractures, respectively. MATERIALS AND METHODS: In our department, all patients with clinically suspected scaphoid fractures and normal scaphoid series of radiographs undergo magnetic resonance imaging (MRI) of the wrist. We selected 50 cases with unequivocal MRI evidence of scaphoid fracture, 50 cases with distal radius fracture and 50 cases with no MRI evidence of bony injury. All 150 initial plain radiographs were examined retrospectively in random order without knowledge of the MRI findings and the scaphoid and pronator fat stripes scrutinized. RESULTS: The scaphoid fat stripe was abnormal in only 25 cases (50%) with confirmed scaphoid fracture on MRI. The pronator fat stripe was abnormal in 13 cases (26%) with confirmed distal radius fracture. In the 50 cases with no MRI evidence of bony injury, the scaphoid fat stripe and pronator fat stripe were abnormal in 25 (50%) and 15 (30%) cases, respectively. The sensitivity and specificity for an abnormal scaphoid fat stripe was 50%. The sensitivity and specificity for an abnormal pronator fat stripe was 26 and 70%, respectively. CONCLUSION: Scaphoid and pronator fat stripes are poor predictors of the presence or absence of underlying occult fractures. PMID- 14521891 TI - Routine MR examination of the knee using parallel imaging. AB - AIM: To demonstrate the feasibility and the clinical value of parallel imaging of the knee. METHODS: A pair of flex coils was used for the parallel imaging. Before optimization of the protocol, qualitative and quantitative analysis was performed by volunteer and phantom study. Two separate consecutive groups of 80 patients were examined with or without parallel imaging and compared with the arthroscopic results. RESULTS: Parallel imaging obtained a total of 164 images of six series in less than 20 min. Compared with conventional imaging, the parallel imaging appeared slightly noisy with an 83% relative signal-to-noise ratio. Demarcation of the cruciate ligaments and menisci were rated equivalent. For the patients study, the parallel imaging delineated ligament and meniscus pathologies with accuracies of over 93%. CONCLUSION: Parallel MR imaging of the knee enables significant reduction of scan time as well as preserving diagnostic results. PMID- 14521892 TI - The need for radio-opaque side markers on CT of the paranasal sinuses. AB - AIM: To audit the presence of a radio-opaque "side marker" routinely placed on the right side of the face of all patients undergoing computed tomography (CT) of the paranasal sinuses in our hospital and to audit awareness levels amongst ear, nose and throat (ENT) clinical and theatre staff regarding the "marker". MATERIALS AND METHODS: An audit of 100 CT studies of the paranasal sinuses was carried out with regard to the presence or absence of the marker. A further audit was conducted relating to the awareness amongst ENT clinical staff and theatre nurses regarding the marker. Re-audit of a second cohort of 100 sinus CT studies was carried out after implementing changes indicated by the first audit cycle. Awareness amongst staff concerned was similarly re-audited. RESULTS: In audit cycle I the side marker was positively identified in 85% of the CT examinations and were deemed absent in 15%. Of the clinical staff interviewed, only 30% were aware of the presence of the "marker" and it's significance. In audit cycle II the "side marker" was positively identified in 100% of the scans reviewed. Awareness of the presence of the "marker" amongst clinical staff was found to be 90%. CONCLUSIONS: The routine placement of a radio-opaque side marker on the right zygoma of all patients undergoing CT of paranasal sinuses is a simple yet effective additional means of denoting the correct side whilst interpreting such images. This has important implications regarding surgery. The audit has shown an improvement in both the placement of the side marker as well as in awareness levels amongst staff involved to ensure best practice. PMID- 14521893 TI - Computed tomography-guided percutaneous catheter drainage of primary and secondary iliopsoas abscesses. AB - AIM: To report our experience with computed tomography (CT)-guided percutaneous catheter drainage (PCD) of iliopsoas abscesses. MATERIALS AND METHODS: Twenty-two iliopsoas abscesses in 21 patients (11 women, 10 men) aged between 18 and 66 years (mean 36 years) were treated with PCD. Abdominal CT demonstrated the iliopsoas abscesses, which were definitively determined by Gram staining and aspirate cultures. Twenty of the 22 iliopsoas abscesses were primary and two were secondary. All PCD procedures were performed under local anaesthesia using a single-step trocar technique (n=19) or Seldinger technique (n=3). RESULTS: PCD was an effective treatment in 21 out of the 22 iliopsoas abscesses. Recurrence was seen in three abscesses as minimal residual collections. Two of them resolved spontaneously with anti-tuberculous regimen. One required percutaneous needle aspiration. The procedure failed in a diabetic patient with a secondary abscess, who died due to sepsis. The length of time that catheters remained in place ranged from 21 to 75 days (mean 59.7 days). Complications included catheter dislocation in four abscesses, which required removal of dislocated catheters and indwelling new ones. CONCLUSION: CT-guided PCD is a safe and effective front-line treatment of iliopsoas abscesses. Surgery should be reserved for failure of PCD and presence of contraindications to PCD. PMID- 14521894 TI - Subendocardial fat: an unusual finding. PMID- 14521895 TI - Imaging features of disseminated peritoneal hydatidosis before and after medical treatment. PMID- 14521896 TI - Gastrostomy insertion. PMID- 14521898 TI - Out of hours percutaneous nephrostomy. PMID- 14521902 TI - New patterns of inheritance in mitochondrial disease. AB - With the identification of a patient with mutated mitochondrial DNA (mtDNA) of paternal origin, it has been unequivocally proven that not only does paternal mtDNA survive in the zygote, but it can also contribute substantially to the mtDNA pool of adult, human skeletal muscle. The questions are: how often does paternal mtDNA inheritance occur and what mechanisms are involved? In this paper, we will review current knowledge on the fate of sperm mitochondria after fertilization and discuss the impact paternal inheritance may have on our understanding of mitochondrial biology. PMID- 14521903 TI - CYP330A1 and CYP4C39 enzymes in the shore crab Carcinus maenas: sequence and expression regulation by ecdysteroids and xenobiotics. AB - Cytochrome P450 enzymes (CYP enzymes) catalyse important metabolic reactions of exogenous and endogenous substrates, including steroid hormones. Here, we report the first two CYP sequences from the shore crab, Carcinus maenas. Two complete cDNAs isolated from crab hepatopancreas encode CYP enzymes named CYP330A1, the first member of a new family, and CYP4C39. CYP330A1 is closest related to members of the CYP2 family (37.3% identical to mouse CYP2J6) and CYP4C39 is most identical to crayfish CYP4C15 (59.5%). CYP330A1 gene expression was induced in hepatopancreas of male green intermoult crabs by ecdysone and ponasterone A, but also by benzo(a)pyrene and phenobarbital. CYP330A1 induction was not observed in red crabs. The present results indicate that the CYP330A1 enzyme may be involved in ecdysteroid metabolism, presumably catabolism, and in the detoxification of environmental pollutants. Ecdysteroids or xenobiotics did not affect CYP4C39 gene expression. The fact that both ecdysteroids and xenobiotics affect CYP330A1 gene expression indicates that mutual interactions between chemical exposures and endocrine functions may exist in the shore crab. PMID- 14521904 TI - Alkylphenol endocrine disrupters inhibit IP3-sensitive Ca2+ channels. AB - We have investigated the influence of alkylphenol endocrine disrupters and the synthetic estrogen diethylstilbestrol (DES) on inositol-1,4,5-trisphosphate (IP(3))-sensitive Ca(2+) channels from porcine cerebellum and rat testicular membranes. All alkylphenols and DES inhibited the extent of IP(3)-induced Ca(2+) release (IICR) from both cerebellar and testicular microsomes. 4-n-nonylphenol was the most potent compound tested (IC(50), 8 microM). Inhibition of IICR was directly related to the length and hydrophobicity of the alkylphenol side chain. None of the alkylphenols or DES appeared to influence the concentration dependence of IICR nor did they have a significant effect on [3H]IP(3) binding to the membranes. An investigation of the effects of nonylphenol on the transient kinetics of IICR showed that it inhibited the rate constants for both the fast and the slow phases of IICR and also the extent of Ca(2+) release. These results illustrate another mechanism by which these environmental pollutants can disrupt endocrine function without the involvement of estrogen receptors. PMID- 14521905 TI - Functional interplay between modulation of histone deacetylase activity and its regulatory role in G2-M transition. AB - The acetylation status of histones plays an essential role in regulating transcription and replication, and is thus involved in the proliferation and differentiation of normal and neoplastic cells. Here, we investigated the effect of trichostatin A (TSA), an inhibitor of histone deacetylases (HDACs), on G2-M transition during the cell cycle. HDAC inhibition by TSA arrested the cell cycle at G2 and also induced escape from the mitotic arrest into G1. TSA reduced the expression of cyclin B1, a key cyclin for G2-M transition, but stimulated expression of p21(WAF1/Cip1), an inhibitor of CDK and Cdc2. In contrast, the expression of cyclin B1 but not p21(WAF1/Cip1) is enhanced during M. Moreover, histone acetylation at promoters of these two genes was regulated by TSA. TSA augmented acetylation of the p21(WAF1/Cip1) promoter but reduced that of the cyclin B1 promoter, suggesting the relationship between TSA-induced modulation of histone acetylation and differential expression of these genes. Taken together, our observations suggest that modulation of HDAC activity is implicated in the G2 M transition by regulating the transcription of cell cycle regulators, p21(WAF1/Cip1) and cyclin B1, via modulating acetylation status of the histones at their promoters. PMID- 14521906 TI - Secretory phospholipase A2 is released from pancreatic beta-cells and stimulates insulin secretion via inhibition of ATP-dependent K+ channels. AB - The release of sPLA(2) from single mouse pancreatic beta-cells was monitored using a fluorescent substrate of the enzyme incorporated in the outer leaflet of the plasma membrane. Stimulation of beta-cells with agents that increased cytosolic free Ca(2+) concentration ([Ca(2+)](i)) induced a rapid release of sPLA(2) to the extracellular medium. Exogenous sPLA(2) strongly stimulated insulin secretion in mouse pancreatic islets at both basal and elevated glucose concentrations. The stimulation of insulin secretion by sPLA(2) was mediated via inhibition of ATP-dependent K(+) channels and an increase in [Ca(2+)](i). Measurements of cell capacitance in single beta-cells revealed that sPLA(2) did not modify depolarisation-induced exocytosis. Our data suggest that a positive feedback regulation of insulin secretion by co-released sPLA(2) is operational in pancreatic beta-cells and point to this enzyme as an autocrine regulator of insulin secretion. PMID- 14521907 TI - Distribution of potential type II restriction sites (palindromes) in prokaryotes. AB - Restriction-modification systems are used as a defensive mechanism against inappropriate invasion of foreign DNA. The recognition sequences for the common type II restriction enzymes and their corresponding methylases are usually palindromes. In this study, we identified the most over- and underrepresented words in DNA of four bacteria: Escherichia coli, Bacillus subtilis, Clostridium perfringens, and Pseudomonas aeruginosa. Using maximum order Markov chain analysis, we found that palindromic words were most often more underrepresented than their non-palindromic counterparts. No strict rule for the intragenic palindrome content could be derived, but for three of the bacteria there was a weak correlation between codon usage bias and palindrome content. A clear drop in palindrome counts was observed in the Shine-Dalgarno region for B. subtilis and C. perfringens, but not in E. coli or P. aeruginosa. It was also shown that palindromes in eubacteria and archaebacteria seem to occur slightly more infrequently than expected on the basis of the genomic GC-content, but some exceptions to this principle exist. PMID- 14521908 TI - Calcium bursts induced by nanosecond electric pulses. AB - We report here real-time imaging of calcium bursts in human lymphocytes exposed to nanosecond, megavolt-per-meter pulsed electric fields. Ultra-short (less than 30 ns), high-field (greater than 1 MV/m), electric pulses induce increases in cytosolic calcium concentration and translocation of phosphatidylserine (PS) to the outer layer of the plasma membrane in Jurkat T lymphoblasts. Pulse-induced calcium bursts occur within milliseconds and PS externalization within minutes. Caspase activation and other indicators of apoptosis follow these initial symptoms of nanosecond pulse exposure. Pulse-induced PS translocation is observed even in the presence of caspase inhibitors. Ultra-short, high-field, electroperturbative pulse effects differ substantially from those associated with electroporation, where pulses of a few tens of kilovolts-per-meter lasting a few tens of microseconds open pores in the cytoplasmic membrane. Nanosecond pulsed electric fields, because their duration is less than the plasma membrane charging time, develop voltages across intracellular structures without porating the cell. PMID- 14521910 TI - Age-related changes of Alzheimer's disease-associated proteins in cynomolgus monkey brains. AB - We characterized senile plaques (SPs) immunohistochemically in cynomolgus monkey brains and also examined age-related biochemical changes of Alzheimer's disease (AD)-associated proteins in these brains from monkeys of various ages. In the neocortex of aged monkeys (>20 years old), we found SPs but no neurofibrillary tangles (NFTs). Antibodies against beta-amyloid precursor protein (APP) or apolipoprotein E (ApoE) stained SPs; however, the pattern of immunostaining was different for the two antigens. APP was present only in swollen neurites, but ApoE was present throughout all parts of SPs. Western blot analysis revealed that the pattern of APP expression changed with age. Although full-length APP695 protein was mainly expressed in brains from young monkeys (4-years-old), the expression of full-length APP751 protein was increased in brains from older monkeys (>20 years old). Biochemical analyses also showed that levels of various AD-associated proteins increased significantly with age in nerve ending fractions. Both SP-associated (APP) and NFT-associated proteins (tau, activated glycogen synthase kinase 3beta, cyclin dependent kinase 5, p35, and p25) accumulated in the nerve ending fraction with increasing age; however, we found no NFTs or paired helical filaments of tau in aged cynomolgus monkey brains. This age-related accumulation of these proteins in the nerve ending fraction was similar to that observed in our laboratory previously for presenilin-1 (PS-1). The accumulation of these SP-associated proteins in this fraction may be a causal event in the spontaneous formation of SPs; thus, SPs may be formed initially in nerve endings. Taken together, these results suggest that intensive investigation of age-related changes in the nerve ending and in axonal transport will contribute to a better understanding of the pathogenesis of neurodegenerative disorders such as AD. PMID- 14521909 TI - A novel diacylglycerol acyltransferase (DGAT2) is decreased in human psoriatic skin and increased in diabetic mice. AB - Psoriasis is a skin disease with epidermal keratinocyte hyperproliferation and altered differentiation. To identify novel psoriasis-related genes, we investigated differentially expressed genes between normal and psoriatic skin. We identified a novel acyl CoA:diacylglycerol acyltransferase 2 (DGAT2) gene, which was decreased in human psoriatic skin. DGAT2 mRNA was expressed in sebaceous glands of normal human skin. DGAT2 protein was detected on endoplasmic reticulum. DGAT2 catalyzes the final step in the production of triglycerides and the accumulation of triglycerides in the tissues is considered to be related to insulin resistance. Therefore, we also investigated the expression of the DGAT2 gene in diabetic mice. DGAT2 mRNA was increased in the adipose, small intestine, and skeletal muscle in diabetic mice. PMID- 14521911 TI - Expression of histone acetyltransferases was down-regulated in poly(ADP-ribose) polymerase-1-deficient murine cells. AB - NF-kappaB-dependent, as well as human immunodeficiency virus type-1 (HIV-1) long terminal repeat (LTR)-dependent, reporter gene expression was significantly impaired in cells derived from poly(ADP-ribose) polymerase-1 (PARP-1)-knockout (PARP-1 -/-) mice. In addition, the level of protein acetylation was markedly lower in PARP-1 -/- cells than control (PARP-1 +/+) cells. Surprisingly, the expression levels of histone acetyltransferases (HATs), p300, cAMP response element-binding protein-binding protein (CBP), and p300/CBP-associated factor (PCAF), were significantly reduced in PARP-1 -/- cells, as compared with PARP-1 +/+ cells. These results suggest that PARP-1 is required for the proper expression of particular HATs. Since p300 and CBP are coactivators of NF-kappaB, we propose here that PARP-1 participates in NF-kappaB-dependent transcription by means of maintaining the expression of HATs. PMID- 14521912 TI - Regulation and differential expression of the c-maf gene in differentiating cultured cells. AB - The Maf transcription factors are involved in a variety of developmental and cellular differentiation processes, but their role in the differentiation of mesenchymal cells has not been described. Here, we have analyzed c-maf expression during the differentiation of adipocytes and muscle cells in cultured systems. The expression of c-maf mRNA was down-regulated during adipogenesis and up regulated during myogenesis. In adipogenesis, the c-maf mRNA was down-regulated 58h after switching to the differentiation medium and just after PPARgamma2 mRNA was induced. A transient transfection analysis of a reporter gene containing the 5(')-flanking region of the c-maf gene showed that PPARgamma2 represses c-maf gene expression. We previously found that c-Maf, c-Jun, and Pax6 bind to and stimulate the c-maf gene. The PPARgamma2 repression of c-maf expression seems to be due, at least in part, to inhibition of the transactivation functions of c Maf, c-Jun, and Pax6. The repression of c-maf was partly reversed by CBP, suggesting that these transcription factors compete for CBP or related transcription co-factors. In myogenesis, there was a differentiation-dependent stimulation of c-maf mRNA expression. The increased expression correlated with myoD expression. A transient transfection analysis showed that myoD stimulated a c-maf reporter gene through binding to two typical E-box elements located between 160 and 180 nucleotides upstream of the cap site. Binding of MyoD to the E-boxes was confirmed by a gel mobility shift assay and DNaseI footprinting analysis. Combined, these results suggest that the c-maf gene plays an important role during the differentiation of adipocyte and muscle cells from mesenchymal fibroblast cells. PMID- 14521913 TI - The significance of N-linked glycosylation in pig endogenous retrovirus infectivity. AB - The significance of the envelope glycoprotein in the transmission of pig endogenous retrovirus (PERV) to human cells was investigated. Pig endothelial cells (PEC) were transduced with the LacZ gene by a pseudotype infection and then infected with PERV subtype B. Culture supernatants of the infected PEC previously incubated with several types of drugs were inoculated into HEK293 cells. The inoculated cells were then stained and the number of LacZ-positive foci was counted. PERV from tunicamycin treated PEC was not transmitted to human cells, indicating the importance of N-linked sugars in this process. Moreover, while inhibition of the terminal alpha-glucose residues from the precursor N-glycan by castanospermine and 1-deoxynojirimycin attenuated PERV infectivity, the mannosidase inhibitors, 1-deoxymannojirimycin and swainsonine, upregulated the infectivity. In addition, treatment with alpha-mannosidase and incubation with concanavalin A completely abrogated the transmission of PERV to HEK293. These data imply that the high-mannose type of N-glycan plays a key role in PERV infectivity. PMID- 14521914 TI - Potentiating effect of distant sites in non-phosphorylated cyclic peptide antagonists of the Grb2-SH2 domain. AB - Without the presence of a phosphotyrosyl group, a phage library derived non phosphorylated cyclic peptide ligand of Grb2-SH2 domain attributed its high affinity and specificity to well-defined and highly favored interactions of its structural elements with the binding pocket of the protein. We have disclosed a significant compensatory role of the Glu(2-) sidechain for the absence of the phosphate functionality on Tyr(0) in the peptide ligand, cyclo(CH(2)CO-Glu(2-) Leu-Tyr(0)-Glu-Asn-Val-Gly-Met(5+)-Tyr-Cys)-amide (termed G1TE). In this study, we report the importance of hydrophobic residue at the Tyr+5 site in G1TE. Both acidic and basic amino acid substitutes are disfavored at this position, and replacement of Met with beta-tert-butyl-Ala was found to improve the antagonist properties. Besides, the polarity of the cyclization linkage was implicated as important in stabilizing the favored binding conformation. Oxidation of the thioether linkage into sulfoxide facilitated the binding to Grb2-SH2 markedly. Simultaneous modification of the three distant sites within G1TE provided the best agent with an IC(50) of 220 nM, which is among the most potent non phosphorous- and non-phosphotyrosine-mimic containing Grb2-SH2 domain inhibitors yet reported. This potent peptidomimetic provides a novel template for the development of chemotherapeutic agents for the treatment of erbB2-related cancer. Biological assays on G1TE(Gla(2-)) in which the original residue of Glu(2-) was substituted by gamma-carboxyglutamic acid (Gla) indicated that it could inhibit the interaction between activated GF receptor and Grb2 protein in cell homogenates of MDA-MB-453 breast cancer cells at the 2 microM level. More significantly, both G1TE(Gla(2-)) alone and the conjugate of G1TE(Gla(2-)) with a peptide carrier can effectively inhibit intracellular association of erbB2 and Grb2 in the same cell lines with IC(50) of 50 and 2 microM, respectively. PMID- 14521915 TI - Identification of four sites of stimulated tyrosine phosphorylation in the MUC1 cytoplasmic tail. AB - MUC1 is an integral membrane protein expressed on the apical surface of epithelial cells where it acts as a signaling receptor. Its cytoplasmic tail (CT) contains seven, highly conserved tyrosine residues, some of which are constitutively phosphorylated and serve as recognition sites for SH2 domain proteins involved in intracellular signal transduction. However, no studies have determined which MUC1 tyrosines are phosphorylated or which signaling pathways are activated in response to stimulation of its ectodomain. In this report, we used our previously characterized CD8/MUC1 chimeric protein that is tyrosine phosphorylated on the MUC1 CT in response to extracellular treatment with CD8 antibody and performed site-directed mutagenesis of all seven tyrosines, both individually and in multiple combinations, to identify the particular sites of stimulated phosphorylation. We observed four phosphorylation sites, three present in sequence motifs with known signaling potential (Y(20), Y(46), and Y(60)) and one previously uncharacterized (Y(29)). These results are discussed in the context of the role of MUC1 in signal transduction. PMID- 14521916 TI - Cell cycle- and activation-dependent regulation of cyclosporin A-induced T cell apoptosis. AB - The immunosuppressive agent cyclosporin A (CsA), which interferes with signal transduction pathways leading to cytokine gene transcription in activated T cells, was investigated regarding its ability to induce apoptosis in T cells undergoing cell cycle progression and activation. In Jurkat and peripheral CD4+ T cells, CsA was found to markedly induce apoptosis at the G0 phase of the cell cycle. Susceptibility to CsA-induced apoptosis progressively decreased during cell cycle progression to the S and G2/M phase, and subsequent T cell receptor- and mitogen-mediated activation totally abrogated CsA-induced apoptosis. Because CsA is an inhibitor of the chymotryptic peptidase activity of the proteasome, susceptibility to apoptosis induced by the proteasome inhibitor lactacystin was investigated under the same conditions. A progressive increase of the susceptibility of T cells to lactacystin-induced apoptosis during cell cycle progression and activation was demonstrated. Intracellular protein levels of the cyclin-dependent kinase inhibitor p27(Kip1)decreased from the G0 to G2/M phase and from the cycling to the activation state, but remained unchanged during the induction of apoptosis by CsA and lactacystin, suggesting a role of p27(Kip1)in the regulation of susceptibility to apoptosis during cell cycle progression and activation. Inhibition of CsA- but not lactacytin-induced apoptosis by overexpression of Bcl-2 in Jurkat T cells revealed that CsA and proteasome inhibitors activate different apoptotic pathways, while both CsA- and lactacystin induced apoptosis were found to be dependent on caspase activation and independent of the FasL/Fas system. The results show that T cells can progressively regulate their susceptibility to apoptosis during cell cycle progression and activation in a stimulus-dependent manner, and suggest that lactacystin, but not CsA, is able to deplete activated T cells by apoptosis, a mechanism deemed necessary for the induction of allograft tolerance. PMID- 14521917 TI - Shear stress induces apoptosis in vascular smooth muscle cells via an autocrine Fas/FasL pathway. AB - Endothelial lesions may lead to the exposure of vascular smooth muscle cells (VSMCs) to the blood flow. In such circumstances VSMCs are exposed to shear stress, an extraordinary mechanical stimulus for this type of cells. Rat VSMCs are cultivated in normal tissue culture plates (statically) or in a cone-plate viscometer (dynamically). Dynamic cultivation leads to a great increase of apoptosis. Immunofluorescence reveals the shear-stress-dependent expression of fas. Apoptosis can be induced by addition of fas ligand-a process which can be blocked by antibodies against either fas or fas ligand. Conditioned medium of dynamically cultivated VSMCs contains fas ligand as the only active apoptosis inducing activity. Apoptosis can be blocked by caspase inhibitors. So the exposure of VSMCs to shear stress leads to apoptosis by the establishment of an autocrine loop of fas and fas ligand-a potential mechanism for the prevention of narrowing of vessel diameter by VSMC proliferation. PMID- 14521918 TI - Two-dimensional crystallization of a small heat shock protein HSP16.3 on lipid layer. AB - As a member of small heat shock proteins, HSP16.3 was identified as the major membrane-bound protein of Mycobacterium tuberculosis during stationary phase. Previous studies revealed that HSP16.3 was in a nonameric form in solution. Here, two-dimensional crystal of HSP16.3 molecules on lipid monolayer was obtained for the first time. The crystal exhibited p422 symmetry with lattice parameters a=b=90A, gamma=90 degrees. The projection map of untilted crystals showed that the basic unit of the crystal was a rod-like structure with two high-density regions. The three-dimensional map at 2.2 nm resolution revealed a rod-like structure with a dimension of 56A x 32A x 25A, similar to the dimeric forms of M. jannaschii HSP16.5 and wheat HSP16.9. Cross-linking experiments confirmed that HSP16.3 nonamers dissociated into dimers upon interaction with the positively charged lipid layer. Surface plasmon resonance measurements revealed that both electrostatic and hydrophobic forces involved in the formation of the 2D crystal on the lipid monolayer. These results provide a basis for further investigation on the unique dimeric structure of HSP16.3 and its functions in vivo. PMID- 14521919 TI - Concomitant production of nitric oxide and superoxide in human macrophages. AB - Many harmful effects of nitric oxide are caused by the reaction of NO with superoxide anion. The present study was carried out to find out the concomitant production of superoxide and to investigate a suitable inhibitor of NO, which is produced by iNOS. THP-1 cells were differentiated into macrophages by PMA and cytokine. Addition of L-NAME showed decrement in superoxide production. Addition of apocynin, aminoguanidine or ONO 1714 brought about a significant reduction in superoxide production. The expressions of p67 and p47(phox) were reduced by the addition of apocynin, aminoguanidine or ONO 1714 whereas xanthine oxidase and cyclooxygenase did not have a major role in superoxide production. The results of the present study show that iNOS and NADPH oxidase play an important role in superoxide release. It suggests that addition of iNOS inhibitor together with apocynin may be more effective in case of therapeutic application in disease conditions like atherosclerosis. PMID- 14521920 TI - Red wine polyphenols cause endothelium-dependent EDHF-mediated relaxations in porcine coronary arteries via a redox-sensitive mechanism. AB - Moderate consumption of wine is associated with cardiovascular protection most likely by increasing the endothelial formation of nitric oxide (NO). The present study investigated whether red wine polyphenolic compounds (RWPCs) increase the formation of endothelium-derived hyperpolarizing factor (EDHF) in arteries and, if so, to characterize the underlying mechanism. Porcine coronary artery rings were suspended in organ chambers for measurement of changes in isometric tension and membrane potential in the presence of indomethacin and N(omega)-nitro-L arginine. RWPCs caused pronounced endothelium-dependent relaxations and hyperpolarizations, which were reduced by the combination of charybdotoxin plus apamin (two inhibitors of EDHF-mediated responses). Both responses to RWPCs were also reduced by antioxidants, membrane permeant analogues of superoxide dismutase, and diphenylene iodonium, an inhibitor of flavin-dependent enzymes. RWPCs induced the formation of superoxide in cultured endothelial cells. These findings demonstrate that RWPCs cause EDHF-mediated relaxations of coronary arteries, which are critically dependent on a redox-sensitive mechanism involving a flavin-dependent enzyme. PMID- 14521922 TI - Identification of histone deacetylase-3 domains that interact with the orphan nuclear receptor TR2. AB - The orphan nuclear receptor TR2 interacts directly with histone deacetylase HDAC3 and HDAC4. We now report that two domains of HDAC3 are involved in its interaction with TR2. GST pull-down assays show that both the N-terminal (residues 1-135) and the C-terminal (residues 210-428) segments of HDAC3 directly interact with TR2. The interaction is also demonstrated in coimmunoprecipitation experiments. The two TR2-binding sites of HDAC3 compete with each other for binding to TR2. The two receptor-interacting domains (RIDs) of HDAC3 were further dissected and mapped to amino acid residues 1-70 and 270-320. In vivo studies demonstrate that HDAC3 and TR2 can form a complex on the TR2 DNA target and this complex exhibits histone deacetylase activity. These data identify two RIDs of HDAC3 and the biological activity of the complex formed by TR2 and HDAC3 on the TR2 DNA target. PMID- 14521921 TI - A novel macrocyclic tetrapeptide mimetic that exhibits low-picomolar Grb2 SH2 domain-binding affinity. AB - The growth factor receptor-bound protein 2 (Grb2) is an SH2 domain-containing docking module that participates in the signaling of numerous oncogenic growth factor receptor protein-tyrosine kinases (PTKs). Presented herein is a 5 methylindolyl-containing macrocyclic tetrapeptide mimetic (5) that binds to Grb2 SH2 domain protein with K(d)=75 pM. This represents the highest affinity yet reported for a synthetic inhibitor against any SH2 domain. In whole cell assays this novel analogue is able to effectively block the association of Grb2 to cognate cytoplasmic erbB-2 at IC(50)<10nM without prodrug derivatization or the addition of carrier peptide motifs. Anti-mitogenic effects against erbB-2 dependent breast cancers are achieved at non-cytotoxic concentrations (IC(50)=0.6 microM). Macrocycle 5 may be representative of a new class of therapeutically relevant Grb2 SH2 domain-directed agents. PMID- 14521923 TI - The p38 MAPK inhibitor, PD169316, inhibits transforming growth factor beta induced Smad signaling in human ovarian cancer cells. AB - Transforming growth factor beta (TGFbeta) can signal through a variety of Smad independent pathways, including the p38 MAPK pathway. Recent work has shown that inhibitors of p38 MAPK, such as SB203580 and SB202190, can inhibit signaling induced by TGFbeta. Here we show that another p38 MAPK inhibitor, PD169316, abrogates signaling initiated by both TGFbeta and Activin A, but not bone morphogenetic protein (BMP) 4. Inhibition of TGFbeta signaling is dose dependent and results in reduced Smad2 and Smad3 phosphorylation, nuclear translocation, and up-regulation of the TGFbeta target gene Smad7. Reduced TGFbeta signaling is not due to abrogation of p38 MAPK activity, since blocking p38 MAPK activity with a dominant negative form of p38 MAPK has no effect on TGFbeta/Smad signaling. Our results show that use of PD169316 at 5 MICROM or higher can block TGFbeta signaling activity and thus caution must be used when attributing cellular activities exclusively to p38 MAPK signaling when these inhibitors are used experimentally. PMID- 14521924 TI - Apoptosis-associated tyrosine kinase is a Cdk5 activator p35 binding protein. AB - A 3(')-terminal fragment of a splice variant of KIAA0641, a human homologue of apoptosis-associated tyrosine kinase (AATYK), was screened from human brain cDNA libraries by a yeast two-hybrid system using a Cdk5 activator p35 as a bait. The cloned cDNA encoded 477 amino acids, composed of internal 458 amino acids of KIAA0641 and 19 amino acids unique to this variant after splicing, then referred to this clone as hAATYKs-p35BP (human AATYK short isoform-p35 binding polypeptide). Using GST-fusion protein, hAATYKs-p35BP was shown to bind to Cdk5/p35 in a rat brain extract. hAATYKs made by fusing the kinase domain of KIAA0641 to the N-terminus of hAATYKs-p35BP was used for binding to Cdk5/p35 in HEK293 cells. Both hAATYKs and KIAA0641 bound to and were phosphorylated by Cdk5/p35. These results suggest that both isoforms of hAATYK are novel Cdk5/p35 binding and substrate proteins. PMID- 14521925 TI - Role of Ca2+ signaling in initiation of stretch-induced apoptosis in neonatal heart cells. AB - Abnormal mechanical load, as seen in hypertension, is found to induce heart cell apoptosis, yet the signaling link between cell stretch and apoptotic pathways is not known. Using an in vitro stretch model mimicking diastolic pressure stress, here we show that Ca(2+) signaling participates essentially in the early stage of stretch-induced apoptosis. In neonatal rat cardiomyocytes, the moderate 20% stretch resulted in tonic elevation of intracellular free Ca(2+) ([Ca(2+)](i)). Buffering [Ca(2+)](i) by EGTA-AM, suppressing ryanodine-sensitive Ca(2+) release, and blocking L-type Ca(2+) channels all prevented the stretch-induced apoptosis as assessed by phosphatidylserine exposure and nuclear fragmentation. Notably, Ca(2+) suppression also prevented known stretch-activated apoptotic events, including caspase-3/-9 activation, mitochondrial membrane potential corruption, and reactive oxygen species production, suggesting that Ca(2+) signaling is the upstream of these events. Since [Ca(2+)](i) did not change without activating mechanosensitive Ca(2+) entry, we conclude that stretch-induced Ca(2+) entry, via the Ca(2+)-induced Ca(2+) release mechanism, plays an important role in initiating apoptotic signaling during mechanical stress. PMID- 14521926 TI - Small heat shock protein Hsp16.3 modulates its chaperone activity by adjusting the rate of oligomeric dissociation. AB - Small heat shock proteins usually exist as oligomers and appear to undergo dynamic dissociation/reassociation, with oligomeric dissociation being a prerequisite for their chaperone activities. However, contradictory cases were also reported that chaperone activities could be enhanced with no change or even increase in oligomeric sizes. Using Hsp16.3 as a model system, our studies show the following: (1) Although a preheat (over 60 degrees C) treatment or the presence of low concentrations of urea (around 0.8M) hardly caused any change in the oligomeric size of Hsp16.3 proteins when examined by size exclusion chromatography, its chaperone activities were increased significantly. (2) Further analysis using the unique pore-gradient polyacrylamide gel electrophoresis revealed a dramatic increase in the tendency of oligomeric dissociation for both the preheated and urea-containing Hsp16.3. (3) Meanwhile, for both cases, an apparent increase in the rate constants of oligomeric dissociation was also observed, as determined by utilizing conjugated fluorescence probes whose quantum yield increases accompanying oligomeric dissociation. (4) Moreover, the fluorescence anisotropy analysis also demonstrated that the oligomeric structures for the preheated or urea-containing Hsp16.3 proteins seem to be more dynamic and variable. In light of these observations, we propose that the small heat shock proteins like Hsp16.3 can modulate their chaperone activities by adjusting the rate of oligomeric dissociation in responding to environmental changes. Results obtained here also suggest that small heat shock proteins might be able to "remember" their stress experiences via certain structural alterations which will allow them to act as better chaperones when the stress conditions reappear. PMID- 14521928 TI - Identification of a ligand for glucocorticoid-induced tumor necrosis factor receptor constitutively expressed in dendritic cells. AB - Glucocorticoid-induced tumor necrosis receptor (GITR) has been implicated in regulation of T cell suppression by CD25(+)CD4(+) regulatory T cells (Tregs). We isolated a cDNA encoding GITR ligand (GITRL) from mouse endothelioma cells. When stably expressed in HEK293 cells, its specific interaction with GITR was confirmed by flow cytometry with the use of GITR-Fc. The interaction was greatly diminished by the addition of soluble GITRL. Consistent with this, soluble GITRL bound to the cell surface of the GITR-expressing HEK293 cells. Coexpression of GITR with GITRL or stimulation of the GITR-expressing cells with soluble GITRL led to activation of NF-kappaB, which was significantly reduced by anti-GITR. More importantly, GITRL was expressed by both immature and mature dendritic cells, suggesting that the interaction between GITR and GITRL may contribute to immune regulation of Tregs by dendritic cells. This isolated TNFRL represents a bona fide GITRL whose presence has been elusive until this time. PMID- 14521929 TI - Inhalant nitrite exposure alters mouse hepatic angiogenic gene expression. AB - Inhalant nitrites are drugs of abuse that have been shown to enhance tumor growth rate in mice and are epidemiologically linked to an increased risk of Kaposi's sarcoma. Because nitrites produce nitric oxide, we hypothesized that their toxicological effects might be partly mediated via regulation of angiogenic factors such as vascular endothelial growth factor (VEGF). Preliminary studies showed that isobutyl nitrite (ISBN) incubation stimulated VEGF protein expression in J774 macrophage cells. C57BL/6 mice exposed to ISBN in air exhibited significant up-regulation of VEGF protein and mRNA in the liver, but not in the lung. Liver mRNA expression of VEGF receptor 2 (VEGFR-2), VEGFR-3, Smad5, and Smad7 was also significantly altered. These results demonstrate that in vivo exposure to an inhalant nitrite results in altered tissue expression of VEGF and its receptors, suggesting that some of its toxicological effects may be mediated partly through a mechanism involving angiogenesis. PMID- 14521927 TI - Androgen regulation of the human FERM domain encoding gene EHM2 in a cell model of steroid-induced differentiation. AB - We have developed a cell model to investigate steroid control of differentiation using a subline of HT1080 cells (HT-AR1) that have been engineered to express the human androgen receptor. Dihydrotestosterone (DHT) treatment of HT-AR1 cells induced growth arrest and cytoskeletal reorganization that was associated with the expression of fibronectin and the neuroendocrine markers chromogranin A and neuron-specific enolase. Expression profiling analysis identified the human FERM domain-encoding gene EHM2 as uniquely induced in HT-AR1 cells as compared to 16 other FERM domain containing genes. Since FERM domain proteins control cytoskeletal functions in differentiating cells, and the human EHM2 gene has not been characterized, we investigated EHM2 steroid-regulation, genomic organization, and sequence conservation. We found that DHT, but not dexamethasone, induced the expression of a 3.8 kb transcript in HT-AR1 cells encoding a 504 amino acid protein, and moreover, that human brain tissue contains a 5.8 kb transcript encoding a 913 amino acid isoform. Construction of an unrooted phylogenetic tree using 98 FERM domain proteins revealed that the human EHM2 gene is a member of a distinct subfamily consisting of nine members, all of which contain a highly conserved 325 amino acid FERM domain. PMID- 14521931 TI - Effect of C-terminal truncations on MLK7 catalytic activity and JNK activation. AB - Mixed lineage kinase 7 (MLK7) is a MAPKKK with enriched expression in heart and skeletal muscle that functions to activate JNK and p38. The MLKs have several conserved domains, including a leucine zipper that in other family members mediates oligomerization critical for catalytic activity and JNK activation. Nested C-terminal deletion mutants of MLK7 from 436 to 286 as well as a mutant lacking only the leucine zipper (delLZ) were generated to determine the role of these domains in catalytic activity and JNK activation. Specific activity of MLK7366 was 75% full length while 436, 322, and delLZ retained approximately 25% and 286, 4% of the full-length catalytic function, demonstrating that the leucine zipper, while not absolutely necessary for catalytic activity, is required to reach full catalytic function of the enzyme. Co-transfection studies of JNK with the MLK7 mutants demonstrated full JNK activation with MLK7, 436, and delLZ, marginal activation for 1-400 or 1-366, and no activation for 1-322, demonstrating that the leucine zipper is not required for JNK activation and that sequence contained in C-terminal residue 322-436 is necessary for full pathway activation by MLK7. PMID- 14521930 TI - Glucose and galactose regulate intestinal absorption of cholesterol. AB - A dose-dependent increase in cholesterol absorption was induced by glucose addition (0-75 mM) to the apical medium of TC7 cells, a well-characterized clone of Caco-2. The uptake into the cells and the secretion rate to the basolateral space were both enhanced by glucose and galactose. This up-regulation was suppressed by SGLT1 inhibition but not by GLUT2 inhibition. Cholesterol cell uptake was significantly decreased by PMA and increased by chelerythrine, with more pronounced changes in the presence of hexoses. Thus, the involvement of a protein kinase C signalling pathway was evidenced in the regulation processes of intestinal cholesterol absorption. In the presence of antibodies directed to hSR BI cholesterol absorption was reduced by 40% and glucose or galactose no longer enhanced it. We suggest that glucose or galactose, through an interaction with SGLT1, activates a protein kinase C pathway that regulates the activity of one of the intestinal cholesterol transporters, namely hSR-BI. PMID- 14521932 TI - Promoter-specific function of the TATA element in undifferentiated P19 cells. AB - P19 embryonal carcinoma cells differentiate into neuronal cells when treated with retinoic acid (RA). To explore the importance of core promoter structures in the regulation of gene expression during neuronal differentiation, the activities of three classes of modified or unmodified model promoters (Spec2a, OtxE, and Ars) were compared in P19 cells before and after RA treatment. The Spec2a promoter was activated in undifferentiated cells specifically when the E-box was located at a proximal position, whereas the OtxE promoter was activated when the E-box was in a distal position. The Ars promoter was only slightly activated by this element. In addition, the TATA element reduced the level of activation provided by the E box, but only when it was located in the Spec2a core promoter. These results indicate that the core promoter structure may govern, at least in part, the stage specific expression of endogenous genes involved in the neuronal differentiation of P19 cells. PMID- 14521933 TI - Hepatocyte nuclear factor 1 negatively regulates amylin gene expression. AB - Maturity-onset diabetes of the young (MODY) is a monogenic subtype of Type 2 diabetes, defined as having an early age of onset, with a dominant inheritance pattern. Hepatocyte nuclear factor 1 (HNF1), which is encoded by the MODY3 gene, has been shown to bind the insulin promoter. Since the promoters of three pancreas-specific genes involved in glucose homeostasis-insulin, glucokinase, and amylin bind similar transcription factors, we were interested in whether HNF1 could also regulate amylin expression. In the present study, we used the electrophoretic mobility shift assay, to demonstrate that the HNF1 transcription factor can specifically bind to the amylin promoter. Moreover, co-transfection of an HNF1 expression vector with an amylin-CAT reporter plasmid decreased the activity of the amylin promoter by 85%. These data support the hypothesis that the amylin gene is regulated by HNF1 in a negative manner and may explain partially how HNF1 mutations result in diabetes. PMID- 14521934 TI - Angiotensin II induction of AP-1 in neurons requires stimulation of PI3-K and JNK. AB - Angiotensin II (Ang II) acts via its type 1 (AT(1)) receptor in neurons to regulate the activity of multiple intracellular signaling molecules, including intracellular Ca(2+), protein kinase C, phosphatidylinositol 3-kinase (PI3-K), and c-Jun NH(2)-terminal kinase (JNK). The present studies investigated the upstream signaling molecules involved in the Ang II stimulation of activator protein-1 (AP-1) DNA binding in neurons. Treatment of neurons cultured from neonatal rat hypothalamus and brainstem with Ang II (100 nM) showed a time dependent increase in AP-1 DNA binding and this effect was inhibited by the AT(1) receptor antagonist, losartan (1 microM), the PI3-K inhibitor, LY294002 (10 microM), and the JNK inhibitor, JNK inhibitor II (100 nM). Furthermore, Ang II (100 nM) causes a time-dependent increase in JNK activity which was attenuated by PI3-K inhibition. These data establish, for the first time, a signaling cascade involved in the Ang II activation of AP-1 DNA binding in neurons. PMID- 14521935 TI - Crystal structure of the platelet activator convulxin, a disulfide-linked alpha4beta4 cyclic tetramer from the venom of Crotalus durissus terrificus. AB - Convulxin (CVX), a C-type lectin, isolated from the venom of the South American rattlesnake Crotalus durissus terrificus, causes cardiovascular and respiratory disturbances and is a potent platelet activator which binds to platelet glycoprotein GPVI. The structure of CVX has been solved at 2.4A resolution to a crystallographic residual of 18.6% (R(free)=26.4%). CVX is a disulfide linked heterodimer consisting of homologous alpha and beta chains. The heterodimers are additionally linked by disulfide bridges to form cyclic alpha(4)beta(4)heterotetramers. These domains exhibit significant homology to the carbohydrate-binding domains of C-type lectins, to the factor IX-binding protein (IX-bp), and to flavocetin-A (Fl-A) but sequence and structural differences are observed in both the domains in the putative Ca(2+)and carbohydrate binding regions. PMID- 14521936 TI - Preliminary 2-D chromatographic investigation of the human stem cell proteome. AB - Stem cells represent a promising tool for the treatment of various hematopoietic diseases. In order to identify stem cell-specific proteins, the proteome of human stem cells from umbilical cord blood was explored for the first time. For this purpose, the crude lysate of 4 x 10(5) CD34+ cells was subjected to in solution trypsin digestion. The resulting peptides were then separated via cation exchange followed by reversed phase chromatography and analyzed by nanospray MS/MS. Database search revealed a total of 215 proteins which could be reliably identified. To obtain a more complete picture of the human stell cell proteome and to also access low abundant proteins, pooling of more than one CD34+ preparations seems necessary in order to increase the cell number and thus the protein content. PMID- 14521937 TI - Involvement of reactive oxygen species and SP-1 in fibronectin production by oxidized LDL. AB - We examined the mechanisms responsible for the production of fibronectin induced by oxidized low-density lipoprotein (oxLDL) in rat mesangial cells. oxLDL accelerated the production of fibronectin with the preceding generation of reactive oxygen species (ROS). Pretreatment with N-acetylcysteine suppressed the oxLDL-induced fibronectin production as well as ROS generation. oxLDL also elicited the activation of SP-1, nuclear factor-kappaB, and cAMP response element binding protein, but not activator protein-1. Among these activated transcription factors, N-acetylcysteine inhibited the activation of SP-1 only. 7 Ketocholesterol, an oxidized lipid in oxLDL particles, induced the production of fibronectin and the activation of SP-1, those which were suppressed by N acetylcysteine. Furthermore, mithramycin A, an inhibitor of SP-1, also suppressed the oxLDL- and 7-ketocholesterol-stimulated production of fibronectin. These results suggest that oxLDL stimulates fibronectin production, at least in part, through the ROS-dependent activation of SP-1 in rat mesangial cells, and further that the ROS-dependent cellular responses may be elicited by 7-ketocholesterol. PMID- 14521938 TI - Expression in a RabGAP yeast mutant of two human homologues, one of which is an oncogene. AB - The yeast proteins Msb3p and Msb4p are two Ypt/Rab-specific GTPase-activating proteins (GAPs) involved in cell growth polarization. Both proteins share with a wide variety of other proteins the highly conserved TBC domain forming the catalytically active RabGAP domain. In particular, Msb3p and Msb4p are similar to the human proteins oncTre210p (the 786-amino-acid product of the human Tre2 oncogene, implicated in Ewing's sarcoma) and RN-tre (a Rab5-GAP controlling endocytosis of the EGFR). To further understand the biochemical function of Tre2 oncogene, we expressed its cDNA and, as a control, the RN-tre cDNA, in an msb3 msb4 double mutant yeast strain. Complementation data show that RN-tre can, unlike Tre2, replace the function of the MSB3 and MSB4 genes. As two highly conserved amino acids, including the catalytic arginine, are mutated in the oncTre210p TBC domain, we restored these two amino acids and expressed the modified Tre2 cDNA in the yeast mutant. PMID- 14521939 TI - Hoechst 33258 binds to G-quadruplex in the promoter region of human c-myc. AB - In vitro binding of Hoechst 33258 to the promoter region of human c-myc, d(GG GGAGGG TGG GGA GGG TGG GGA AGG TGG GG) which forms G-quadruplex, both in vitro and in vivo in the presence of metal ions, was investigated by equilibrium absorption, fluorescence, and kinetic surface plasmon resonance methods. Hypochromic effect in UV absorption spectra and blue shift in fluorescence emission maxima of Hoechst in the presence of quadruplex revealed that Hoechst binds to the quadruplex. Analysis of UV and fluorescence titration data revealed that Hoechst binds to quadruplex with binding affinity of the order of 10(6). Anisotropy measurements and higher lifetime obtained from time-resolved decay experiments revealed that quadruplex-bound Hoechst is rotationally restricted in a less polar environment than the bulk buffer medium. From surface plasmon resonance studies, we obtained kinetic association (k(a)) and dissociation (k(d)) of 1.23+/-0.04 x 10(5)M(-1)s(-1) and 0.686+/-0.009 s(-1), respectively. As Hoechst is known to bind A-T-rich region of duplex DNA, here we propose the likelihood of Hoechst interacting with the AAGGT loop of the quadruplex. PMID- 14521940 TI - Elevated levels of beta defensin-1 mRNA in diabetic kidneys of GK rats. AB - We previously described the isolation of CDK4, representing the 3(')end of an unknown mRNA, using differential display [Biophys. Res. Commun. 232 (1997) 49]. To determine the identity of the CDK4 gene, full-length CDK4 cDNA clones were isolated from a diabetic kidney cDNA library. DNA sequencing revealed that CDK4 represents the rat beta defensin-1 gene (rBD-1). rBD-1 mRNA was detected in rat kidney, heart, lung, and skeletal muscle using RT-PCR. Using Northern blot analysis, elevated levels of the 0.5kb mRNA transcript were detected in the kidneys of 6-, 16-, 26-, and 40-week-old GK rats when compared with age-matched Wistar controls. A novel 2.0kb mRNA transcript was also detected in all kidneys examined from GK rats in none of the age-matched control kidneys. Biglycan and TGF-beta1 mRNAs were significantly up-regulated in kidneys of GK rats at 26 weeks compared to 16 and 6 weeks, showing that the kidneys of GK rats mimic the gene expression pattern described for human and experimental DN. These data suggest that overexpression of beta-defensin-1 mRNA may play a role in diabetic nephropathy. PMID- 14521941 TI - Human AP endonuclease possesses a significant activity as major 3'-5' exonuclease in human leukemia cells. AB - Apurinic/apyrimidinic (AP) endonuclease (Ape1) is the major cellular enzyme responsible for repairing AP-sites in DNA. It can cleave the DNA phosphodiester backbone immediately 5(') to an AP-site. Ape1 also shows 3(')-phosphodiesterase activity, a 3(')-phosphatase activity, and an RNaseH activity. However, regarding its exonuclease activity, it remains controversial whether human Ape1 may possess a 3(')-5(') exonuclease activity. During the course of study to search for the major nuclease activity to double-stranded DNA in human leukemia cells, we purified a 37 kDa Mg(2+)-dependent exonuclease from cytosolic fraction of human leukemia U937 cells. Surprisingly, this exonuclease is Ape1. We demonstrated for the first time that Ape1 possesses a significant activity as major 3(')-5(') exonuclease in human leukemia cells. In addition, we also observed that translocation of cytoplasmic Ape1 into nucleus occurs during DNA damage. PMID- 14521942 TI - Role of class I and class II histone deacetylases in carcinoma cells using siRNA. AB - The role of the individual histone deacetylases (HDACs) in the regulation of cancer cell proliferation was investigated using siRNA-mediated protein knockdown. The siRNA for HDAC3 and HDAC1 demonstrated significant morphological changes in HeLa S3 consistent with those observed with HDAC inhibitors. SiRNA for HDAC 4 or 7 produced no morphological changes in HeLa S3 cells. HDAC1 and 3 siRNA produced a concentration-dependent inhibition of HeLa cell proliferation; whereas, HDAC4 and 7 siRNA showed no effect. HDAC3 siRNA caused histone hyperacetylation and increased the percent of apoptotic cells. These results demonstrate that the Class I HDACs such as HDACs 1 and 3 are important in the regulation of proliferation and survival in cancer cells. These results and the positive preclinical results with non-specific inhibitors of the HDAC enzymes provide further support for the development of Class I selective HDAC inhibitors as cancer therapeutics. PMID- 14521943 TI - Inhibition of authentic hepatitis C virus replication by sodium stibogluconate. AB - Using a hepatitis C virus (HCV) subgenomic RNA replicon system, drugs currently being used to treat other human diseases were examined for their antiviral activities against HCV. Several drugs including sodium stibogluconate, a compound used to treat leishmaniasis, were capable of suppressing replication of HCV replicon. The antiviral effect of sodium stibogluconate was subsequently verified using a cell line (293EBNA-Sip-L) previously proved to be permissive for HCV infection/replication. An ex vivo assay using fresh human liver slices established and a panel of human liver slices was obtained from biopsy samples of patients infected with HCV was used to examine the antiviral activity of this drug. A nearly complete suppression effect was achieved in four of six human liver slices at the drug concentration of 100 microg/ml, lower than what was required to treat leishmaniasis. A human trial is mandatory to understand its clinical value in treating chronic hepatitis C. PMID- 14521944 TI - Activation of signaling pathways by putative scavenger receptor class A (SR-A) ligands requires CD14 but not SR-A. AB - Macrophage scavenger class A type I and type II receptors (SR-A) are trimeric, integral membrane glycoproteins that bind an unusually broad array of macromolecular ligands. These ligands include modified proteins and lipoproteins, nucleic acids, and a variety of plant and microbial cell wall constituents, such as fucoidan and lipoteichoic acid. Early studies of SR-A functions indicated that the receptors bound, internalized, and degraded their ligands without provoking any macrophage activating signaling events. More recent studies have provided evidence that several SR-A ligands can activate macrophage gene expression via utilization of a receptor-linked, PI3-kinase pathway. To investigate the role of SR-A in engaging signal transduction events, we employed macrophages taken from mice lacking these receptors. Using either fucoidan or lipoteichoic acid, we confirm that both ligands stimulate tyrosine phosphorylation of PI3-kinase and production of modest levels of the cytokine, TNFalpha. However, macrophages taken from SR-A null mice did not differ from wild type macrophages in these responses, indicating that these signaling events arise independently of SR-A activity. Employing mice lacking CD14, a GPI anchored receptor that binds bacterial lipopolysaccharide and signals via activation of Toll-like receptors, we show that the fucoidan and lipoteichoic acid responses are largely abrogated when CD14 is absent. These data do not provide support for direct SR-A involvement in signal transduction events and suggest that the early characterization of these receptors as initiators of a non-phlogistic, pathogen clearance pathway was correct. PMID- 14521945 TI - Interleukin-6 production by contracting human skeletal muscle: autocrine regulation by IL-6. AB - Interleukin-6 (IL-6) is a cytokine with immuno-regulatory functions. However, contracting skeletal muscle expresses and subsequently releases IL-6 in high amounts, and recent evidence in IL-6 deficient mice suggests a role of IL-6 in metabolism. Since IL-6 mRNA levels also increase in abdominal adipose tissue in response to exercise, we wanted to examine the possible existence of a positive feedback mechanism between muscle and adipose tissue. We obtained biopsies from human skeletal muscle and abdominal subcutaneous adipose tissue in relation to either 3h of bicycle exercise or recombinant human IL-6 infusion (rhIL-6) or saline infusion. In muscle, IL-6 mRNA increased (p<0.05) immediately after exercise, peaking at this time-point, whereas IL-6 mRNA in adipose tissue increased 1.5-h post exercise (p<0.05) displaying a different kinetic of induction. During rhIL-6 infusion, IL-6 mRNA increased 120-fold in muscle (p<0.05). In conclusion, the present study demonstrates that muscle IL-6 is regulated by an autocrine mechanism at the transcriptional level. PMID- 14521946 TI - Trypanosoma rangeli uptakes the main lipoprotein from the hemolymph of its invertebrate host. AB - During its life cycle Trypanosoma rangeli crosses the hemolymph of its invertebrate host. In the present study, we demonstrate for the first time the uptake of lipophorin (Lp), the main lipid-transporting particle of insect hemolymph. We observed that living T. rangeli parasites uptake lipids from both 32P- and 3H-, or 125I-labeled Lp. However, the parasites do not uptake any other hemolymphatic protein such as 32P-labeled vitellogenin. The presence of a specific receptor to Lp in the parasite surface is suggested based on experiments using 125I-Lp. We also investigated the intracellular fate of lipids using Texas Red-labeled phosphatidylethanolamine-Lp. Parasites were observed under confocal microscope and displayed fluorescent-labeled lipids close to the flagellar pocket and in vesicles at the posterior region. In conclusion, this study raises a novel set of molecular events which takes place during vector-parasite interaction. PMID- 14521947 TI - Conjugated linoleic acid enhances plasma adiponectin level and alleviates hyperinsulinemia and hypertension in Zucker diabetic fatty (fa/fa) rats. AB - Adiponectin is a recently discovered hormone secreted by adipocytes that has been reported to enhance insulin sensitivity. Although insulin resistance and/or compensatory hyperinsulinemia are considered to be involved with hypertension in obese humans, the relationship between plasma adiponectin level and obesity related hypertension has not been fully clarified. In this study, we investigated the effect of dietary conjugated linoleic acid (CLA), reported as an insulin sensitizer, on plasma adiponectin, plasma insulin, and blood pressure in Zucker diabetic fatty (ZDF) rats. During the onset of obesity, blood pressure increased in ZDF rats. The increase, however, was prevented by dietary CLA. After 8 weeks, accumulated plasma insulin and glucose were also attenuated by CLA feeding. Dietary CLA increased plasma adiponectin levels in ZDF rats and the increase was attributed to the enhanced mRNA expression in white adipose tissue. This study provides the first evidence that dietary CLA increases plasma adiponectin level through the enhancement of mRNA expression. We speculate that the increase alleviates hyperinsulinemia and prevents the onset of hypertension in CLA-fed ZDF rats. PMID- 14521948 TI - C-terminal pro-ghrelin peptides are present in the human circulation. AB - We provide the first evidence for the existence in human plasma of peptides derived from the 66 carboxyl-terminal amino acids of pro-ghrelin (C-ghrelin). C ghrelin immunoreactivity in plasma was higher than ghrelin, and did not significantly correlate with body mass index in normal health. In patients with myocardial infarction, plasma levels of both ghrelin and C-ghrelin were significantly decreased (approximately 30%, P<0.05), whereas in patients with heart failure, C-ghrelin levels were significantly elevated (approximately 32%, P<0.05) compared with controls. HPLC coupled with RIA showed circulating C ghrelin to be primarily of low molecular weight (M(r) approximately 3500), but in chronic heart failure, a higher molecular weight form (M(r) approximately 7500) is also present. This is the first evidence for potential circulating hormones derived from the carboxyl terminus of pro-ghrelin and for their modulation in cardiovascular diseases. PMID- 14521949 TI - Comparison of glycosyltransferase families using the profile hidden Markov model. AB - In order to investigate the relationship between glycosyltransferase families and the motif for them, we classified 47 glycosyltransferase families in the CAZy database into four superfamilies, GTS-A, -B, -C, and -D, using a profile Hidden Markov Model method. On the basis of the classification and the similarity between GTS-A and nucleotidylyltransferase family catalyzing the synthesis of nucleotide-sugar, we proposed that ancient oligosaccharide might have been synthesized by the origin of GTS-B whereas the origin of GTS-A might be the gene encoding for synthesis of nucleotide-sugar as the donor and have evolved to glycosyltransferases to catalyze the synthesis of divergent carbohydrates. We also suggested that the divergent evolution of each superfamily in the corresponding subcellular component has increased the complexities of eukaryotic carbohydrate structure. PMID- 14521950 TI - Dynamic compression of cartilage constructs engineered from expanded human articular chondrocytes. AB - Recent works have shown that mechanical loading can alter the metabolic activity of chondrocytes cultured in 3D scaffolds. In this study we determined whether the stage of development of engineered cartilaginous constructs (expanded adult human articular chondrocytes/Polyactive foams) regulates the effect of dynamic compression on glycosaminoglycan (GAG) metabolism. Construct maturation depended on the culture time (3-14 days) and the donor (4 individuals). When dynamic compression was subsequently applied for 3 days, changes in GAG synthesized, accumulated, and released were significantly positively correlated to the GAG content of the constructs prior to loading, and resulted in stimulation of GAG formation only in the most developed tissues. Conversely, none of these changes were correlated with the expression of collagen type II mRNA, indicating that the response of chondrocytes to dynamic compression does not depend directly upon the stage of cell differentiation, but rather on the extracellular matrix surrounding the cells. PMID- 14521951 TI - Gene expression profiling in adipose tissue indicates different transcriptional mechanisms of liver X receptors alpha and beta, respectively. AB - The nuclear receptors liver X receptors (LXR) alpha and beta are important regulators of genes involved in lipid, cholesterol, and carbohydrate metabolism and are highly expressed in mature adipocyte tissue. In this study we show that LXRalpha and LXRbeta are more expressed in brown adipose tissue and subcutaneous white adipose tissue than visceral (gonadal) adipose tissue. Furthermore, we report differences between LXRalpha and LXRbeta in their ability to alter expression of target genes. Gene expression profiling analysis of gonadal white adipose tissue from LXRalpha(-/-) mice and LXRbeta(-/-) mice shows different gene expression patterns in the two LXR-deficient mouse strains. Genes regulated similarly in both KO mouse strains as well as genes regulated in one, but not the other LXR-deficient mouse strain were seen. A number of genes were regulated in opposite directions by the respective LXR isoform. Taken together this suggests that the LXR isoforms might operate through different transcriptional mechanisms as well as common mechanisms. These results are in consonance with the growing body of evidence reporting differences in regulation of gene expression between the two isoforms. Furthermore, gene expression profiling shows altered gene expression patterns in primary mouse embryonic fibroblasts (MEFs) from wild type versus LXRbeta(-/-) mice; MEFs are pluripotent cells with the potential to differentiate into mature adipocytes. These results indicate a role of LXR in early developmental stages of adipose tissue. PMID- 14521952 TI - Differential regulation of secretory leukocyte protease inhibitor and elafin by progesterone. AB - Elafin and secretory leukocyte protease inhibitor (SLPI) are anti-protease and anti-microbial molecules present at mucosal surfaces. Both molecules are expressed in the female reproductive tract where they may be involved in innate immune defence. This study examines the role of progesterone in the regulation of SLPI and elafin. Progesterone treatment increases expression of SLPI mRNA and protein in the T47D breast epithelial cell line and this upregulation is attenuated in the presence of the anti-gestogens, RU486 and ZK98734, confirming the involvement of the nuclear progesterone receptor. A putative progesterone response element has been identified in the SLPI promoter. Progesterone also acts in synergy with the proinflammatory cytokines, IL-1beta and TNFalpha, to increase SLPI. In contrast, progesterone treatment has no direct effect on elafin mRNA expression. In summary, progesterone has a differential effect on SLPI and elafin expression and although both vary within the uterus throughout the menstrual cycle, progesterone is likely to contribute to the direct regulation of SLPI in the female reproductive tract even in the presence of inflammatory agents. PMID- 14521953 TI - A role for CBS domain 2 in trafficking of chloride channel CLC-5. AB - CLC-5 is a member of the CLC family of voltage-gated chloride channels. Mutations disrupting CLC-5 lead to Dent's disease, an X-linked renal tubular disorder, characterised by low molecular weight proteinuria, hypercalciuria, nephrocalcinosis, and renal stones. Sequence analysis of CLC-5 reveals a 746 amino acid protein with an intracellular amino-terminus, transmembrane spanning domains, and two CBS domains within its intracellular carboxy-terminus. CBS domains have been implicated in intracellular targetting and trafficking as well as protein-protein interactions. We investigate subcellular localisation of three naturally occurring CLC-5 mutants which all lead to a truncated protein, disrupting the second CBS domain. These mutants are unable to traffic normally to acidic endosomes but are retained in perinuclear compartments, colocalising with the Golgi complex. This is the first identification of the cellular pathogenesis of CBS domain mutations of CLC-5. PMID- 14521954 TI - Secondary structure analyses of protein films on gold surfaces by circular dichroism. AB - In order to analyze the secondary structures of protein molecules adsorbed on gold surfaces, circular dichroism (CD) spectra were measured and the secondary structure contents of protein ultra-thin films were estimated quantitatively. A disulfide group was introduced to cytochrome b(562) (cyt.b562), which is a water soluble b-type heme protein. The cyt.b562 molecules self-assembled to form an ultra-thin protein film both on a gold substrate modified with 2,2(') dithiodiacetic acid and on a bare gold surface. CD measurements were carried out both in solution and in air, and these results were compared. The protein denaturation was partially prevented, not only in solution but also in air, by both the modification of the substrate and the introduction of the anchor group to the protein molecule. The secondary structure contents of ultra-thin protein films on flat gold surfaces were observed for the first time both in solution and in air by CD spectra. PMID- 14521955 TI - Proteolytic processing of IGFBP-related protein-1 (TAF/angiomodulin/mac25) modulates its biological activity. AB - Insulin-like growth factor (IGF) binding protein-related protein-1 (IGFBP-rP1) was previously identified as tumor-derived adhesion factor (TAF) secreted from human bladder carcinoma cells. It exhibits growth-stimulatory activity in synergy with insulin or IGFs. In the present study, we found that IGFBP-rP1 was proteolytically cleaved to a two-chain form. The cleavage sequence suggested that a trypsin-like serine proteinase may be responsible for the processing. The cleavage of IGFBP-rP1 led to an almost complete loss of both insulin/IGF-1 binding activity and insulin/IGF-1-dependent growth-stimulatory activity. On the other hand, the cell attachment activity of IGFBP-rP1 was markedly increased by the proteolytic processing. Syndecan-1 was thought to be a cell surface receptor for both intact and cleaved IGFBP-rP1 forms. Although the proteolytic cleavage of IGFBP-rP1 decreased its heparin-binding activity, the cleaved form could bind syndecan-1 efficiently. Thus the proteolytic processing of IGFBP-rP1 seems to modulate its insulin/IGF-dependent and -independent biological functions. PMID- 14521956 TI - Regulation on the expression and N-glycosylation of integrins by N acetylglucosaminyltransferase V. AB - The expressions of integrin alpha5, beta1, and alpha6 were studied in H7721 cells by means of flow cytometric and RT-PCR method after transfected with sense and antisense cDNA of N-acetylglucosaminyltransferase V (GnT-V). The transfected cells were characterized by Northern blot. It was found that the order of expression from high to low was beta1>alpha5>alpha6. Transfection of sense GnT-V up-regulated alpha5 and alpha6, but not beta1 subunit, while antisense GnT-V down regulated alpha5 and beta1, but not alpha6. The alterations of surface integrin subunits were quite compatible with the changes of their mRNAs. Using enzyme labeled lectin analysis, it was shown that alpha5 subunit contained only C(2)C(2) biantennary N-glycan, which was not regulated by sense and antisense GnT-V. In contrast, beta1 subunit contained both biantennary and tri-/tetra-antennary N glycans with GlcNAcbeta1,6Manalpha1,6-branch, and the latter was up- and down regulated by the sense and antisense GnT-V, respectively. Therefore, the amount of biantennary N-glycans on beta1 subunit, but not the integrin protein, was correlated to the cell adhesion to fibronectin and laminin, which was reduced and elevated in the sense and antisense GnT-V-transfected cells, respectively, as we previously reported. PMID- 14521957 TI - EGCG corrects aberrant splicing of IKAP mRNA in cells from patients with familial dysautonomia. AB - Familial dysautonomia (FD) is an autosomal recessive neurodegenerative disorder. The most prevalent causative mutation is a T-->C transition in a donor splice site of the IKBKAP transcript, resulting in aberrant splicing and a truncated protein. The mutation's position and leaky nature suggested that its impact might be moderated by altering the level of splice-regulating proteins. The reported ability of (-)-epigallocatechin gallate (EGCG), a polyphenol, to down-regulate the expression of hnRNP A2/B1, a trans-activating factor that encourages the use of intron-distal 5(') splice sites, prompted an evaluation of its effect on the IKBKAP transcript in FD-derived cells. EGCG reduces the level of hnRNP A2/B1 and increases the amounts of the wild-type IKBKAP-encoded transcript and functional protein. Combined treatment of cells with EGCG and tocotrienol, which upregulates IKBKAP transcription, results in a synergistic production of the functional gene product. These findings suggest the possible use of EGCG as a therapeutic modality for individuals with FD. PMID- 14521958 TI - Differential inhibition of transient outward currents of Kv1.4 and Kv4.3 by endothelin. AB - The effects of endothelin on the transient outward K(+) currents were compared between Kv1.4 and Kv4.3 channels in Xenopus oocytes expression system. Both transient outward K(+) currents were decreased by stimulation of endothelin receptor ET(A) coexpressed with the K(+) channels. Transient outward current of Kv1.4 was decreased by about 85% after 10(-8) M ET-1, while that of Kv4.3 was decreased by about 60%. By mutagenesis experiments we identified two phosphorylation sites of PKC and CaMKII in Kv1.4 responsible for the decrease in I(to) by ET-1. In Kv4.3 a PKC phosphorylation site was identified which is in part responsible for the decrease in I(to). Differences in the suppression of I(to) could be ascribed to the difference in intracellular signaling including the number of phosphorylation sites. These findings might give clues for the understanding of molecular mechanism of ventricular arrhythmias in heart failure, in which endothelin is involved in the pathogenesis. PMID- 14521959 TI - Resistin is expressed in pancreatic islets. AB - Resistin, a recently described adipocyte factor, is regulated by peroxisome proliferator-activated receptor gamma (PPARgamma) agonists. While resistin has been proposed to mediate insulin resistance in rodents, little is known about human resistin and its expression in pancreatic islets has not been tested. The goal of the present study was therefore to analyze whether resistin, like PPARgamma, is expressed in islets. Human islets from seven donors were analyzed by quantitative RT-PCR revealing resistin expression in all samples. Immunohistochemistry using a resistin-specific antibody on human pancreatic sections localized resistin protein to the islets. Mouse resistin was also detected in the Min6 beta cell line. Interestingly, we found a 4-fold increase in islet resistin expression in insulin resistant A-ZIP transgenic compared to wild type mice. Our results demonstrate that resistin is expressed in islets and up regulated in insulin resistance and thereby shed new light on the role of resistin in mice and humans. PMID- 14521961 TI - Modifying the oligomeric state of cyclic amidase and its effect on enzymatic catalysis. AB - A group of cyclic amidases, including hydantoinase, allantoinase, dihydropyrimidinase, and dihydroorotase, catalyze the reversible hydrolysis of cyclic ureides, such as 5-monosubstituted hydantoins and dihydropyrimidines. These four enzymes carry hydrophobic patches to form dimers. With the exception of dihydroorotase, these enzymes are further dimerized to form tetramers by hydrophobic interactions. This leads us to speculate that the hydrophobic interaction domain may be a significant factor in the catalytic property of these oligomeric cyclic amidases, for which activities are not allosterically regulated. We generated a dimeric D-hydantoinase by mutating five residues in the hydrophobic alpha-helical interface of a tetramer and analyzed the kinetic properties of the dimeric form of D-hydantoinase. The specific activity of the dimeric D-hydantoinase corresponds to 5.3% of the activity of tetrameric D hydantoinase. This low specific activity of the dimeric D-hydantoinase indicates that the dimeric interaction to form a tetramer has a significant effect on the catalytic activity of this non-allosteric tetramer. PMID- 14521960 TI - Abnormal signalling of 14-3-3 proteins in cells with accumulated xanthurenic acid. AB - Xanthurenic acid is an endogenous molecule leading to caspase-9 and -3 activation. Here we report that xanthurenic acid targets signalling proteins 14-3 3 into lysosomes leading to interruption protein/protein interaction. Xanthurenic acid changed the localisation of 14-3-3 in the cells. At a concentration of 10 and 20 microM the 14-3-3 was translocated into lysosomes. At these concentrations Bad and cofilin were dephosphorylated. Translocation of dephosphorylated Bad into mitochondria and cytochrome c release were observed. Cofilin dephosphorylation in the presence of xanthurenic acid was associated with lack of the apoptotic actin cytoskeleton disintegration. In conclusion xanthurenic acid accumulation in cells abolished the regulatory function of the proteins 14-3-3 in the cell physiology and caused misfolding of the proteins leading to cell pathology. PMID- 14521962 TI - Targeted nucleotide exchange in the CAG repeat region of the human HD gene. AB - Huntington's disease (HD) is marked by the expansion of a tract of repeated CAG codons in the HD-gene, IT15. Once expressed, the expanded poly Q region of the huntingtin protein (Htt), which is normally soluble, becomes insoluble, leading to the formation of intracellular inclusions and ultimately to neuronal degeneration. Interruption of the pure poly Q tract at the genetic level should undermine the transition from Htt solubility to Htt insolubility. Modified single stranded oligonucleotides were used to direct the nucleotide exchange of an A residue to a T residue in the second codon of the HD-gene, resulting in the creation of a leucine residue among the poly Q tract. Consistent with results from other groups, we provide evidence that short synthetic DNA molecules can modify the HD-gene directly, preliminarily offering a potential therapeutic approach to Huntington's disease. PMID- 14521963 TI - A comprehensive search for HNF-3alpha-regulated genes in mouse hepatoma cells by 60K cDNA microarray and chromatin immunoprecipitation/PCR analysis. AB - To characterize the regulatory pattern by a specific transcription regulatory factor, we used a combination of expression analysis with the mouse cDNA microarray composed of 60,000 cDNA clones and cross-linking/chromatin immunoprecipitation (X-ChIP) followed by comparative PCR. Overexpression of mouse hepatocyte nuclear factor-3alpha (HNF-3alpha) in a mouse hepatoma cell line resulted in accompanied perturbed expression of more than 1500 genes. Search for HNF-3alpha consensus recognition sequences in the upstream regions of their coding sequences, which were mapped on the mouse genome, enabled us to mine 300 genes as the potential HNF-3alpha-regulated genes and classify 135 annotated ones into several functional categories. Further X-ChIP/PCR analysis demonstrated in vivo binding of HNF-3alpha to the 5(')-flanking sequences of 25 members selected out of these genes. Besides known HNF-3alpha-regulated genes such as albumin and alpha-fetoprotein genes, the genes newly identified as the HNF-3alpha-regulated ones include three encoding CDP-diacylglycerol-inositol 3 phosphatidyltransferase, phosphatidylserine decarboxylase, and phospholipase A2, which are located en suite in the lipid metabolic pathway in liver. The potential usefulness of the present approach to extensive characterization of gene expression framework directed by a specific transcription regulatory factor is discussed. PMID- 14521964 TI - Retaining nurses over fifty. PMID- 14521966 TI - Requesting and interpreting trauma radiographs: a role extension for accident & emergency nurses. AB - Government supported expansion of the nursing role within Accident & Emergency (A&E) departments in the United Kingdom (UK) has begun to break down the traditional barriers to professional practice. Today, many nurses working within A&E departments are both requesting and interpreting radiographic examinations as part of their normal working practice. However, role expansion does not occur without increased responsibility. Unsatisfactory requests for radiography and inaccurate radiographic interpretation may result in inappropriate patient treatment, misuse of resources, patient recall and litigation. Nurses undertaking these role extensions need to ensure that their levels of knowledge and skill to perform the role are appropriate and adequately supported. This article summarises the results of a national questionnaire survey of A&E nurse managers that aimed to identify current working practices, including education, training and limitations to practice, with respect to the requesting and interpretation of trauma radiographs by A&E nurses. PMID- 14521965 TI - Avoidable cardiac arrest: lessons for an A&E department. AB - Deterioration to cardiac arrest is not always sudden and unexpected and on a significant number of occasions cardiac arrest could be prevented. This has important messages for the Accident and Emergency (A&E) department as the vast majority of emergency admissions originate via A&E. PMID- 14521967 TI - Sexual health in the emergency department -- what do we need to know? PMID- 14521968 TI - Detecting failed thrombolysis in the accident and emergency department. AB - The primary objective in managing a patient with ST segment elevation myocardial infarction (STEMI) is to establish reperfusion in the infarct-related artery and to maintain it. Two approaches to coronary reperfusion are used in the UK - primary angioplasty and intravenous thrombolysis. Primary angioplasty is the gold standard approach to managing STEMI, but in the UK (due to financial, resource and personnel limitations) this is not the first-line treatment. Thrombolytic therapy remains the most widely used approach and the benefits of such an approach are irrefutable; thrombolysis saves lives, reduces infarct size and limits left ventricular dysfunction. However, data from the thrombolytic trials also suggest that 30-40% of patients fail to reperfuse with standard thrombolytic therapy. Similar data demonstrates that patients who do not sustain adequate perfusion in the infarct-related artery have a poor prognosis and increased mortality rates. As long as thrombolysis remains the standard therapy for STEMI, it is important that patients in whom the treatment has been unsuccessful are swiftly recognised and appropriate interventions instituted. The criteria to assess successful reperfusion of the infarct-related artery need to be simple to apply, easy to interpret and non-invasive. This article will discuss the most useful criteria to make such a diagnosis and suggest approaches to enable recognition of 'failed thrombolysis' in the accident and emergency department. The current views on managing failed thrombolysis will conclude the article. PMID- 14521970 TI - An in-car safety campaign for children in Southern Derbyshire. PMID- 14521969 TI - Nurse triage, diagnosis and treatment of eye casualty patients: a study of quality and utility. AB - INTRODUCTION: Patients presenting to the Accident and Emergency department of a district general hospital with an eye problem are referred directly to a dedicated eye casualty service. They are then triaged by a staff nurse from the eye department. This eye casualty sees around 8000 patients per annum. This study assesses whether the nurse triage of eye casualty patients forms an effective filter of problems which do not require the attention of the doctor on duty. The incidence of cases of misdiagnosis and inappropriate discharge of patients resulting in a delay in diagnosis and treatment was also assessed. METHODS: All patients presenting in a 3-month period managed solely by the triage nurse, were identified from the casualty register, and the hospital records retrieved and examined. RESULTS: Four hundred and forty of a total 1976 patients (22%) were seen exclusively by triage nurses; eight (2.5%) of these 440 patients returned unplanned to the eye department. In all cases it was considered that the return of the patient would not have been preventable by initial attention of the ophthalmologist on duty. DISCUSSION: With appropriate threshold for referral, nurses trained in slit lamp examination can offer a successful service to safely diagnose and treat common eye casualty presentations. PMID- 14521971 TI - The experience of the nurse at triage influences the timing of CPAP intervention. AB - Increasing attention in the last decade has shown that intervention of continuous positive airway pressure therapy (CPAP) in cardiogenic pulmonary oedema (CPO) markedly improves the outcome of patients presenting with acute respiratory failure. This study used a non-experimental correlational design to research the relationship between the experience of the nurse, with the application of CPAP to patients presenting to a metropolitan emergency department with CPO and to establish what difference, if any, CPAP made to outcomes. A retrospective audit of records was used to extract data on all 54 patients that received CPAP over a 12-month period. The primary outcome measures were off CPAP within 2 h, transfer to intensive care unit (ICU) or cardiac care unit (CCU) and secondary outcome measures were length of hospital stay and death. There was a trend towards more experienced nurses attending patients who required immediate treatment or treatment within 10 min. These patients were more likely to be recognised at triage as requiring CPAP therapy. The early application of CPAP reduced hospital mortality, length of stay, and the need for intubation and ventilation. Attention needs to be given on how best to educate nurses so that more patients presenting with acute respiratory failure can benefit from nurses' decision-making regarding the commencement of CPAP. PMID- 14521972 TI - MedicAlert -- more than just a bracelet! PMID- 14521984 TI - The eyes suppress a circadian rhythm of FOS expression in the suprachiasmatic nucleus in the absence of light. AB - Photic information transmitted from the eyes to the suprachiasmatic nucleus (SCN) is essential for entrainment of circadian behavioral and physiological rhythms in mammals. Under conditions of constant darkness, these rhythms are maintained by the circadian pacemaker cells of the SCN [Bioessays 22 (2000) 23]. It is possible, however, that even in the absence of light, the eyes, which also contain autonomous circadian pacemakers [Science 272 (1996) 419; Chronobiol Int 16 (1999) 229], modulate circadian rhythms in the SCN. Indeed, it was shown recently that removal of the eyes abolishes an endogenous circadian rhythm within cells of the SCN [Nat Neurosci 6 (2003) 111], a finding that led to the suggestion that specific rhythms of the SCN are driven by input from the eyes. In contrast, we show here that removal of the eyes amplifies a normally dampened endogenous circadian rhythm within the SCN, indicating that the eyes can suppress the expression of specific rhythms within the SCN while promoting others. PMID- 14521985 TI - Intraseptal infusion of oxotremorine impairs memory in a delayed-non-match-to sample radial maze task. AB - The medial septal nucleus is part of the forebrain circuitry that supports memory. This nucleus is rich in cholinergic receptors and is a putative target for the development of cholinomimetic cognitive-enhancing drugs. Septal neurons, primarily cholinergic and GABAergic, innervate the entire hippocampal formation and regulate hippocampal formation physiology and emergent function. Direct intraseptal drug infusions can produce amnestic or promnestic effects depending upon the type of drug administered. However, intraseptal infusion of the cholinomimetic oxotremorine has been reported to produce both promnestic and amnestic effects when administered prior to task performance. The present study examined whether post-acquisition intraseptal infusion of oxotremorine would be promnestic or amnestic in a delayed-non-match-to-sample radial maze task. In this task rats must remember information about spatial locations visited during a daily sample session and maintain that information over extended retention intervals (hours) in order to perform accurately on the daily test session. Treatments may then be administered during the retention interval. Alterations in maze performance during the test session an hour or more after treatment evidences effects on memory. In the present study, intraseptal infusion of oxotremorine (1.0-10.0 microg) produced a linear dose-related impairment of memory performance. Importantly, we also observed disrupted performance on the day after treatment. This persistent deficit was related only to memory over the retention interval and did not affect indices of short-term memory (ability to avoid repetitive or proactive errors during both the pre- and post-delay sessions). The persistent deficit contrasts with the acute amnestic effects of other intraseptally administered drugs including the cholinomimetics carbachol and tacrine. Thus, intraseptal oxotremorine produced a preferential disruption of memory consolidation as well as a persistent alteration of medial septal circuits. These findings are discussed with regards to multi-stage models of hippocampal-dependent memory formation and the further development of therapeutic strategies in the treatment of mild cognitive impairment as well as age-related decline and Alzheimer's dementia. PMID- 14521986 TI - Stimulant doses of caffeine induce c-FOS activation in orexin/hypocretin containing neurons in rat. AB - Although caffeine is a commonly used CNS stimulant, neuronal mechanisms underlying its stimulatory effect are not fully understood. Orexin (hypocretin) containing neurons play a critical role in arousal and might be activated by acute administration of caffeine. We examined this possibility by using dual immunostaining for orexin B and c-Fos protein as a marker for neuronal activation. Rats were administered intraperitoneally with 10, 30 or 75 mg/kg caffeine, or saline. As previously reported, caffeine increased locomotion at 10 and 30 mg/kg, but not at 75 mg/kg. The numbers of orexin-immunoreactive and non orexin-immunoreactive neurons expressing c-Fos were analysed using three counting boxes within the orexin field in the posterior hypothalamus. Compared with saline, all doses of caffeine increased the number of cells immunoreactive for both orexin and c-Fos. The average magnitude of this increase across doses in orexin neurons differed amongst regions; c-Fos expression increased by 343% in the perifornical area and by 158% in the more medial, dorsomedial nucleus. In the lateral hypothalamic area, c-Fos expression increased by 226% at 10 and 30 mg/kg but no change was seen at 75 mg/kg. In contrast, caffeine significantly increased the number of non-orexin-immunoreactive neurons expressing c-Fos only in the dorsomedial nucleus. These results indicate that systemically administered caffeine preferentially activates orexin neurons over non-orexin neurons in the same field, and that this activation is most pronounced in the perifornical region where orexin neurons are most concentrated. The activation of orexin neurons might play a role in the behavioural activation by caffeine. PMID- 14521987 TI - Neck muscle fatigue affects postural control in man. AB - We hypothesised that, since anomalous neck proprioceptive input can produce perturbing effects on posture, neck muscle fatigue could alter body balance control through a mechanism connected to fatigue-induced afferent inflow. Eighteen normal subjects underwent fatiguing contractions of head extensor muscles. Sway during quiet stance was recorded by a dynamometric platform, both prior to and after fatigue and recovery, with eyes open and eyes closed. After each trial, subjects were asked to rate their postural control. Fatigue was induced by having subjects stand upright and exert a force corresponding to about 35% of maximal voluntary effort against a device exerting a head-flexor torque. The first fatiguing period lasted 5 min (F1). After a 5-min recovery period (R1), a second period of fatiguing contraction (F2) and a second period of recovery (R2) followed. Surface EMG activity from dorsal neck muscles was recorded during the contractions and quiet stance trials. EMG median frequency progressively decreased and EMG amplitude progressively increased during fatiguing contractions, demonstrating that muscle fatigue occurred. After F1, subjects swayed to a larger extent compared with control conditions, recovering after R1. Similar findings were obtained after F2 and after R2. Although such behaviour was detectable under both visual conditions, the effects of fatigue reached significance only without vision. Subjective scores of postural control diminished when sway increased, but diminished more, for equal body sway, after fatigue and recovery. Contractions of the same duration, but not inducing EMG signs of fatigue, had much less influence on body sway or subjective scoring. We argue that neck muscle fatigue affects mechanisms of postural control by producing abnormal sensory input to the CNS and a lasting sense of instability. Vision is able to overcome the disturbing effects connected with neck muscle fatigue. PMID- 14521988 TI - The mitochondrial complex I inhibitor annonacin is toxic to mesencephalic dopaminergic neurons by impairment of energy metabolism. AB - The death of dopaminergic neurons induced by systemic administration of mitochondrial respiratory chain complex I inhibitors such as 1-methyl-4 phenylpyridinium (MPP(+); given as the prodrug 1-methyl-1,2,3,6 tetrahydropyridine) or the pesticide rotenone have raised the question as to whether this family of compounds are the cause of some forms of Parkinsonism. We have examined the neurotoxic potential of another complex I inhibitor, annonacin, the major acetogenin of Annona muricata (soursop), a tropical plant suspected to be the cause of an atypical form of Parkinson disease in the French West Indies (Guadeloupe). When added to mesencephalic cultures for 24 h, annonacin was much more potent than MPP(+) (effective concentration [EC(50)]=0.018 versus 1.9 microM) and as effective as rotenone (EC(50)=0.034 microM) in killing dopaminergic neurons. The uptake of [(3)H]-dopamine used as an index of dopaminergic cell function was similarly reduced. Toxic effects were seen at lower concentrations when the incubation time was extended by several days whereas withdrawal of the toxin after a short-term exposure (<6 h) arrested cell demise. Unlike MPP(+) but similar to rotenone, the acetogenin also reduced the survival of non-dopaminergic neurons. Neuronal cell death was not excitotoxic and occurred independently of free radical production. Raising the concentrations of either glucose or mannose in the presence of annonacin restored to a large extent intracellular ATP synthesis and prevented neuronal cell demise. Deoxyglucose reversed the effects of both glucose and mannose. Other hexoses such as galactose and fructose were not protective. Attempts to restore oxidative phosphorylation with lactate or pyruvate failed to provide protection to dopaminergic neurons whereas idoacetate, an inhibitor of glycolysis, inhibited the survival promoting effects of glucose and mannose indicating that these two hexoses acted independently of mitochondria by stimulating glycolysis. In conclusion, our study demonstrates that annonacin promotes dopaminergic neuronal death by impairment of energy production. It also underlines the need to address its possible role in the etiology of some atypical forms of Parkinsonism in Guadeloupe. PMID- 14521989 TI - Mechanisms underlying the inhibition of long-term potentiation by preconditioning stimulation in the hippocampus in vitro. AB - We have investigated the mechanisms underlying a form of metaplasticity, namely the inhibition by preconditioning stimulation of high frequency stimulation (HFS) induced long-term potentiation (LTP) in the medial perforant path of the dentate gyrus. Preconditioning stimulation (weak 50 Hz) was found to inhibit subsequent LTP induction if applied 10-20 min, but not 2 or 45 min, prior to the HFS. Preconditioning stimulation in the form of low frequency stimulation did not block LTP induction. The inhibition of LTP was not caused by a reduction in N methyl-D-aspartate receptor (NMDAR) transmission, as the preconditioning stimulation did not reduce isolated NMDAR-mediated EPSPs. The involvement of group I and group II metabotropic glutamate receptor (mGluR) activation in the inhibition of LTP was demonstrated by experiments in which the inhibition of LTP by the preconditioning stimulation was prevented by the presence of antagonists of group I or group II mGluR during the preconditioning stimulation. Moreover, group I and group II mGluR agonists directly inhibited subsequent LTP induction. The involvement of NMDAR in the preconditioning stimulation was shown by the ability of an NMDAR antagonist to prevent the inhibition of LTP by the preconditioning stimulation. The preconditioning inhibition of LTP induction was shown by the use of kinase inhibitors to involve activation of PKC and p38 MAP kinase, but not p42 MAP kinase or tyrosine kinase. We conclude that the preconditioning inhibition of LTP induction is a complex process which involves activation of NMDAR, group I and group II mGluR, and intracellular cascades activating PKC and p38 MAP kinase. PMID- 14521990 TI - Dehydroepiandrosterone 7-hydroxylase CYP7B: predominant expression in primate hippocampus and reduced expression in Alzheimer's disease. AB - Neurosteroids such as dehydroepiandrosterone (DHEA), pregnenolone and 17beta estradiol are synthesized by cytochrome P450s from endogenous cholesterol. We previously reported a new cytochrome P450 enzyme, CYP7B, highly expressed in rat and mouse brain that metabolizes DHEA and related steroids by hydroxylation at the 7alpha position. Such 7-hydroxylation can enhance DHEA bioactivity in vivo. Here we show that the reaction is conserved across mammalian species: in addition to mouse and rat, DHEA hydroxylation activity was present in brain extracts from sheep, marmoset and human. Northern blotting using a human CYP7B complementary deoxyribonucleic acid (cDNA) probe confirmed the presence of CYP7B mRNA in marmoset and human hippocampus; CYP7B mRNA was present in marmoset cerebellum and brainstem, with lower levels in hypothalamus and cortex. In situ hybridization to human brain revealed higher levels of CYP7B mRNA in the hippocampus than in cerebellum, cortex, or other brain regions. We also measured CYP7B expression in Alzheimer's disease (AD). CYP7B mRNA was significantly decreased (approximately 50% decline; P<0.05) in dentate neurons from AD subjects compared with controls. A decline in CYP7B activity may contribute the loss of effects of DHEA with ageing and perhaps to the pathophysiology of AD. PMID- 14521991 TI - Apolipoprotein E4 decreases whereas apolipoprotein E3 increases the level of secreted amyloid precursor protein after closed head injury. AB - Apolipoprotein E (apoE4) and head trauma are important genetic and environmental risk factors for Alzheimer's disease. Furthermore, apoE4 increases both the acute and chronic consequences of head trauma. The latter are associated with the deposition of amyloid-beta, which is particularly elevated in apoE4 subjects. The short-term effects of head injury are associated with transiently increased metabolism of amyloid precursor protein (APP) and its secreted fragment, APPs. In the present study, we examined the possibility that the acute, short-term pathological effects of apoE4 following head trauma and the corresponding neuroprotective effects of apoE3 are related to isoform-specific effects of apoE on APP metabolism. Accordingly, male transgenic mice expressing human apoE3 or apoE4 on a null mouse apoE background and apoE-deficient and control mice were subjected to closed head injury (CHI). The resulting effects on brain APP, and on its secreted products, APPs and secreted product of the alpha-cleavage of APP (APPsalpha) were then determined 24 h following injury. Immunoblotting revealed no significant differences between the basal APP, APPs and APPsalpha levels of the hippocampus or the cortex of the control and the apoE3 and ApoE4 transgenic mice. The apoE-deficient mice also had similar cortical basal levels of APP and its metabolites, whereas their corresponding basal hippocampal APP and APPs levels were lower than those of the other groups. CHI lowered the hipppocampal APPs and APPsalpha levels of the apoE4 transgenic mice, whereas those of the apoE3 transgenic mice and of the control and apoE-deficient mice were not affected by this insult. In contrast, CHI raised the cortical APP and APPs levels of the apoE3 transgenic mice but had no significant effect on those of the other mice groups. These animal model findings suggest that the acute, short-term pathological effects of apoE4 following CHI and the corresponding neuroprotective effects of apoE3 may be mediated by their opposing effects on the expression and cleavage of cortical and hippocampal APP. Similar isoform-specific interactions between apoE and APP may play a role in the acute, short-term effects of head trauma in humans. PMID- 14521992 TI - Sex differences in the hippocampal dentate gyrus of the guinea-pig before puberty. AB - The aim of the present research was to ascertain the presence of sex differences in the hippocampal dentate gyrus of the guinea-pig, a long-gestation rodent which gives birth to mature young and whose brain is at a more advanced stage of maturation at birth than that of the rat and mouse. The brains of neonatal (15-16 days old) and prepubescent (45-46 days old) male and female guinea pigs were Golgi-Cox stained. Granule cells were sampled from the upper (suprapyramidal) and lower (infrapyramidal) blade of the septal dentate gyrus and their dendritic tree and soma were measured. The analysis was conducted separately on granule cells with soma in the superficial (superficial granule cells) and deep (deep granule cells) half of the granule cell layer. Numerous sex differences were found in the upper blade of the dentate gyrus. Neonatal males had more dendritic branches than females in the innermost dendritic tree of both superficial and deep granule cells, but females had more branches over the middle/outer dendritic tree and a longer dendritic length. In prepubescent animals, the sex difference in the middle dendritic tree of the superficial granule cells changed direction, with males having more branches than females. In the deep granule cells, the sex differences were similar to those in neonatal animals. In both granule cell types, the dendritic length was similar in the two sexes. While no sex differences were found in dendritic spine density in neonatal animals, in prepubescent animals spine density was greater in females. In the lower blade the granule cells showed very few sex differences in both neonatal and prepubescent animals. This study shows wide dynamically changing sex differences in the granule cells located in the upper blade of the septal dentate gyrus, but almost no differences in the lower blade. These results demonstrate that sex differences are not ubiquitous in the dentate gyrus and suggest that the lower blade, unlike the upper blade, might be involved in non-sexually dimorphic behaviors. PMID- 14521993 TI - Learning deficits in forebrain-restricted brain-derived neurotrophic factor mutant mice. AB - Brain-derived neurotrophic factor (BDNF) participates in synaptic plasticity and the adaptive changes in the strength of communication between neurons thought to underlie aspects of behavioral adaptation. By selectively deleting BDNF from the forebrain of mice using the Cre site-specific DNA recombinase, we were able to study the requirements for BDNF in behaviors such as learning and anxiety. Early onset forebrain-restricted BDNF mutant mice (Emx-BDNF(KO)) that develop in the absence of BDNF in the dorsal cortex, hippocampus, and parts of the ventral cortex and amygdala failed to learn the Morris Water Maze task, a hippocampal dependent visuo-spatial learning task. Freezing during all phases of cued contextual fear conditioning, a behavioral task designed to study hippocampal dependent associative learning, was enhanced. These mice learned a brightness discrimination task well but were impaired in a more difficult pattern discrimination task. Emx-BDNF(KO) mice did not exhibit altered sensory processing and gating, as measured by the acoustic startle response or prepulse inhibition of the startle response. Although they were less active in an open-field arena, they did not show alterations in anxiety, as measured in the elevated-plus maze, black-white chamber or mirrored chamber tasks. Combined, these data indicate that although an absence of forebrain BDNF does not disrupt acoustic sensory processing or alter baseline anxiety, specific forms of learning are severely impaired. PMID- 14521994 TI - The mouse nac1 gene, encoding a cocaine-regulated Bric-a-brac Tramtrac Broad complex/Pox virus and Zinc finger protein, is regulated by AP1. AB - NAC1 cDNA was identified as a novel transcript induced in the nucleus accumbens from rats chronically treated with cocaine. NAC1 is a member of the Bric-a-brac Tramtrac Broad complex/Pox virus and Zinc finger family of transcription factors and has been shown by overexpression studies to prevent the development of behavioral sensitization resulting from repeated cocaine treatment. This paper reports the cloning and characterization of the corresponding gene. The mouse Nac1 gene consist of six exons, with exon 2 containing an alternative splice donor, providing a molecular explanation of the splice variants observed in mouse and rat. Transcripts of Nac1 were ubiquitously detected in different mouse tissues with prominent expression in the brain. The mouse Nac1 gene was localized to chromosome 8, suggesting a highly plausible candidate gene to explain differences in cocaine-induced behaviors between C57BL6/J and DBA/2J mice that had previously been mapped to the area. In addition, a functional AP1 binding site has been identified in an intron 1 enhancer of the Nac1 gene that plays an essential role in the activation of the gene in differentiation of neuroblastoma cells. Co-transfection with c-jun and c-fos expression plasmids, which encode the two subunits of AP1, activated the wild type Nac1 intron 1 enhancer two-fold over basal, nearly at the level of NAC1 enhancer activity seen in differentiated N2A cells. Mutation of the AP1 site completely abrogated all activation of the NAC1 enhancer in differentiated N2A cells. Activation of immediate early genes such as c-fos and c-jun following chronic drug treatments has been well characterized. The present data describe one potential regulatory cascade involving these transcription factors and activation of NAC1. Identification of drug induced alterations in gene expression is key to understanding the types of molecular adaptations underlying addiction. PMID- 14521995 TI - Frequency-dependent expression of corticotropin releasing factor in the rat's cerebellum. AB - Corticotropin releasing factor (CRF), localized in extrinsic afferents in the mammalian cerebellum, is defined as a neuromodulator within cerebellar circuits, and appears to be an essential element in the generation of long term depression, a proposed mechanism for motor learning. These physiological studies are based on exogenous application of CRF and do not address potential mechanisms that may influence endogenous release of the peptide. In the present study, immunohistochemistry was used to analyze changes in the lobular distribution of CRF-like immunoreactivity (LIR). In addition radioimmunoassay (RIA) was used to quantify changes in levels of the peptide in the cerebellum following stimulation of the inferior cerebellar peduncle (ICP) at 10 or 40 Hz or the inferior olivary nucleus (ION) at 1, 5, 10, or 20 Hz. Results indicate that there is a greater distribution of CRF-like-immunolabeled climbing fibers, mossy fibers, and astrocytes in all lobules of the cerebellum that is directly related to stimulation frequency. Maximal effects were elicited with 40 Hz ICP and 5-10 Hz ION stimulation. Quantitatively, the RIA studies indicate that there is a significant increase in CRF levels in the vermis, hemispheres and flocculus that correlates closely with stimulation frequency. In conclusion, stimulation of cerebellar afferents induces a significant change in the distribution and levels of CRF-LIR in climbing fibers, mossy fibers and glial cells. This suggests that the modulatory effects ascribed to CRF may influence a greater number of target neurons when levels of activity in afferent systems is increased. PMID- 14521996 TI - The neuroprotective effect of a novel calmodulin antagonist, 3-[2-[4-(3-chloro-2 methylphenyl)-1-piperazinyl]ethyl]-5,6-dimethoxy-1-(4-imidazolylmethyl)-1H indazole dihydrochloride 3.5 hydrate, in transient forebrain ischemia. AB - A novel calmodulin (CaM) antagonist DY-9760e, (3-[2-[4-(3-chloro-2-methylphenyl) 1-piperazinyl]ethyl]-5,6-dimethoxy-1-(4-imidazolylmethyl)-1H-indazole dihydrochloride 3.5 hydrate), with an apparent neuroprotective effect in vivo, potently inhibits CaM-dependent nitric oxide synthase in situ. In the present study, we determined whether DY-9760e inhibits nitric oxide (NO) production and protein nitration by peroxynitrite (ONOO(-)) formation in the hippocampal CA1 region of gerbils after transient forebrain ischemia. In freely moving gerbils, NO production after 10-minute forebrain ischemia was monitored consecutively with in vivo brain microdialysis. Pretreatment with DY-9760e (50 mg/kg i.p.) significantly decreased the increased levels of NO(x)(-) (NO metabolites, NO(2)( ) plus NO(3)(-)) immediately after, 24 h after cerebral ischemia-reperfusion to the control levels of sham-operated animals. Western blot and immunohistochemical analyses using an anti-nitrotyrosine antibody as a marker of ONOO(-) formation indicated a marked increase in nitrotyrosine immunoreactivity in the pyramidal neurons of the CA1 region 2 h after reperfusion, and DY-9760e significantly inhibited increased nitrotyrosine immunoreactivity. Coincident with the inhibition of the NO production and protein tyrosine nitration, pretreatment with DY-9760e rescued the delayed neuronal death in the hippocampal CA1 region. These results suggest that the inhibitory effects of DY-9760e on the NO-ONOO(-) pathway partly account for its neuroprotective effects in cerebral ischemia. PMID- 14521997 TI - Local mu and delta opioid receptors regulate amphetamine-induced behavior and neuropeptide mRNA in the striatum. AB - The purpose of this study was to investigate the role that mu and delta opioid receptor blockade has upon stimulant-induced behavior and neuropeptide gene expression in the striatum. Acute administration of amphetamine (2.5 mg/kg i.p.) caused an increase in behavioral activity and preprodynorphin, substance P, and preproenkephalin mRNA expression. Intrastriatal infusion of the mu opioid antagonist, H-D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH(2) (CTAP), or the delta opioid antagonist, H-Tyr-Tic[CH(2)NH]-Phe-Phe-OH (TIPPpsi), significantly decreased amphetamine-induced vertical activity. However, only CTAP reduced amphetamine-induced distance traveled. Quantitative in situ hybridization histochemistry revealed that CTAP blocked amphetamine-induced preprodynorphin and substance P mRNA. However, preproenkephalin mRNA levels in the dorsal striatum were increased to the same extent by CTAP, amphetamine, or a combination of the two drugs. In contrast, TIPPpsi significantly decreased amphetamine-induced mRNA expression of all three neuropeptides. These data indicate that both mu and delta receptor subtypes differentially regulate amphetamine-induced behavior and neuropeptide gene expression in the rat striatum. PMID- 14521998 TI - Quantification and localization of kainic acid-induced neurotoxicity employing a new biomarker of cell death: cleaved microtubule-associated protein-tau (C-tau). AB - Previous studies of neuronal degeneration induced by the neurotoxin, kainic acid, employed silver stain techniques that are non-quantitative or ELISA measurement of the non-neuronal protein, glial fibrillary acidic protein. As previous studies employed biomarkers that were either non-quantitative or non-neuronal, the present study employed a new neuronally localized biomaker of neuronal damage, cleaved microtubule-associated protein (MAP)-tau (C-tau). The time course of kainate neurotoxicity was quantitatively determined in several brain regions in the present study employing a C-tau specific ELISA. Differences in ELISA determined regional brain levels of C-tau were compared with the density of somatodendritic C-tau labeling qualitatively determined in immunohistochemical anatomical mapping studies of kainic acid-treated animals. Immunoblot studies revealed that the C-tau antibodies employed in the present study were highly specific for proteolytic cleaved C-tau. Immunolabeling of 45 kD-50 kD C-tau proteins was observed only in brain samples from kainic acid-treated but not vehicle-treated rats. Time course studies revealed that C-tau levels determined by ELISA were maximal 3 days after kainic acid with C-tau levels increasing 26 fold in hippocampus, 16-fold in cortex and four-fold in both striatum and hypothalamus. These statistical differences in maximal C-tau levels observed in the ELISA studies were similar to differences qualitatively observed in C-tau immunohistochemical studies. C-tau immunohistochemistry revealed extensive damage in hippocampal regions CA1 and 3, moderate damage in several cortical regions and mild damage in striatum and hypothalamus. Similar cleavage of rat MAP-tau to C tau has been reported after neuronal degeneration induced by neurotoxic doses of methamphetamine and neuronal degeneration resulting from bacterial meningitis. In humans, C-tau proteolysis has been demonstrated to be a reliable biomarker of neuronal damage in traumatic brain injury and stroke where cerebrospinal C-tau levels are correlated with patient clinical outcome. These data suggest that C tau proteolysis may prove a reliable species independent biomarker of neuronal degeneration regardless of source of injury. PMID- 14521999 TI - Localisation of P2X2 receptor subunit immunoreactivity on nitric oxide synthase expressing neurones in the brain stem and hypothalamus of the rat: a fluorescence immunohistochemical study. AB - A large body of evidence suggests that nitric oxide (NO) and ATP act as neurotransmitters in the regulatory mechanisms concerning several autonomic functions at the level of both the hypothalamus and the brain stem. In the present study, we investigated whether neuronal NO synthase containing neurones also express P2X(2) receptor subunit of the ATP-gated ion channel via double labelling fluorescence immunohistochemistry. Our data demonstrate that a high percentage of neuronal NO synthase-immunoreactive neurones are also P2X(2) immunoreactive in the rostral ventrolateral medulla (98%) and supraoptic nucleus of the hypothalamus (92%). Significant numbers of neuronal NO synthase immunoreactive neurones are also P2X(2)-immunoreactive in the subpostremal (48%) and commissural (65%) subdivisions of the nucleus tractus solitarius. In the caudal ventrolateral medulla and raphe obscurus, 96% and 89%, respectively, of neuronal NO synthase containing neurones also express P2X(2) receptor subunit. In contrast to the supraoptic nucleus, there was a lower percentage of co localisation between NO synthase and P2X(2) receptor subunit in the paraventricular nucleus of the hypothalamus. In summary, this study demonstrates for the first time that there is a widespread co-localisation of neuronal NO synthase and P2X(2) receptor subunit in the hypothalamus and brain stem of the rat. Further studies are required to elucidate whether NO and ATP functionally interact within the hypothalamus and the brain stem. PMID- 14522000 TI - The impact of Abeta-plaques on cortical cholinergic and non-cholinergic presynaptic boutons in alzheimer's disease-like transgenic mice. AB - A previous study in our laboratory, involving early stage, amyloid pathology in 8 month-old transgenic mice, demonstrated a selective loss of cholinergic terminals in the cerebral and hippocampal cortices of doubly transgenic (APP(K670N,M671L)+PSl(M146L)) mice, an up-regulation in the single mutant APP(K670N,M671L) mice and no detectable change in the PSl(M146L) transgenics [J Neurosci 19 (1999) 2706]. The present study investigates the impact of amyloid plaques on synaptophysin and vesicular acetylcholine transporter (VAChT) immunoreactive bouton numbers in the frontal cortex of the three transgenic mouse models previously described. When compared as a whole, the frontal cortices of transgenic and control mice show no observable differences in the densities of synaptophysin-immunoreactive boutons. An individual comparison of layer V of the frontal cortex, however, shows a significant increase in density in transgenic models. Analysis of the cholinergic system alone shows significant alterations in the VAChT-immunoreactive bouton densities as evidenced by an increased density in the single (APP(K670N,M671L)) transgenics and a decreased density in the doubly transgenics (APP(K670N,M671L)+PSl(M146L)). In investigating the impact of plaque proximity on bouton density at early stages of the amyloid pathology in our doubly (APP(K670N,M671L)+PSl(M146L)) transgenic mouse line, we observed that plaque proximity reduced cholinergic pre-synaptic bouton density by 40%, and yet increased synaptophysin-immunoreactive pre-synaptic bouton density by 9.5%. Distance from plaques (up to 60 microm) seemed to have no effect on bouton density; however a significant inverse relationship was visible between plaque size and cholinergic pre-synaptic bouton density. Finally, the number of cholinergic dystrophic neurites surrounding the truly amyloid, Thioflavin-S(+) plaque core, was disproportionately large with respect to the incidence of cholinergic boutons within the total pre-synaptic bouton population. Confocal and electron microscopic observations confirmed the preferential infiltration of dystrophic cholinergic boutons into fibrillar amyloid aggregates. We therefore hypothesize that extracellular Abeta aggregation preferentially affects cholinergic terminations prior to progression onto other neurotransmitter systems. This is supported by the observable presence of non-cholinergic sprouting, which may be representative of impending neuritic degeneration. PMID- 14522001 TI - A 14-day period of hindpaw sensory deprivation enhances the responsiveness of rat cortical neurons. AB - Hypodynamia-hypokinesia (HH) is a model of hindpaw sensory deprivation. It is obtained by unloading of the hindquarters during 14 days. In this situation, the feet are not in contact with the ground and as a consequence, the cutaneous receptors are not activated; the sensory input to the primary somatosensory cortex (SmI) is thus reduced. In a previous study, we have shown that HH induced a cortical reorganisation of the hindlimb representation. The understanding of the mechanisms involved in cortical map plasticity requires a close examination of the changes in response properties of cortical neurons during HH. The aim of the present study was thus to study the characteristics of neurons recorded from granular and infragranular layers in hindlimb representation of SmI. A total of 289 cortical neurons were recorded (158 from control rats and 131 from HH rats) in pentobarbital-anaesthetized rats. Cutaneous threshold, cutaneous receptive fields, spontaneous activity (discharge rate and instantaneous frequency) and activity evoked by air-jet stimulation (response latency and duration, amplitude) were analysed. The present study suggests that activity-dependent changes occur in the cortex. The duration of the spike waveform presented two populations of spikes: thin-spike cells (<1 ms, supposed to be inhibitory interneurons) and regular cells (>1 ms). Thin-spike cells were less frequently encountered in HH than in control rats. The analysis of regular cells revealed that after HH (1) spontaneous activity was unchanged and (2) cortical somatosensory neurons were more responsive: the cutaneous threshold was reduced and the response magnitude increased. Taken together, these results suggest a down-regulation of GABAergic function. PMID- 14522003 TI - Detection and discrimination of carvone enantiomers in rats with olfactory bulb lesions. AB - Rats with lesions of dorsal and dorsolateral bulbar sites known to be differentially responsive to carvone enantiomers were tested for their ability to detect (+)-carvone, to discriminate between (+)-carvone from (-)-carvone, and to discriminate (+)-carvone from mixtures of both enantiomers after they had been pre-trained or not pre-trained on these tasks prior to surgery. In postoperative tests, rats pre-trained on the enantiomer discrimination problems made somewhat fewer errors than those not pre-trained, but experimental rats performed as well as controls (those that had one intact olfactory bulb) within both conditions and on each task. These results indicate that removal of most bulbar sites known to be differentially responsive to carvone enantiomers and the consequent disruption of normal patterns of bulbar input produced in response to carvones are largely without effect on the ability of rats to discriminate between these odors. PMID- 14522002 TI - Expression of auxiliary beta subunits of sodium channels in primary afferent neurons and the effect of nerve injury. AB - Multiple voltage-gated sodium channels are the primary mediators of cell excitability. They are multimers that consist of the pore-forming alpha subunit and auxiliary beta subunits. Although ion permeability and voltage sensing are primarily determined by the alpha subunit, beta subunits are important modulators of sodium channel function. The purpose of this study was to assess the effect of axotomy on the expression of beta subunits (beta(1), beta(2) and beta(3)) and coexpression of Na(v)1.3 and beta(3) subunits in the dorsal root ganglion (DRG). We used sciatic nerve transection models or spared nerve injury (SNI) models in the rat. In reverse transcriptase-polymerase chain reaction analysis, there were no significant differences between contralateral and ipsilateral DRGs of beta(1) and beta(2) mRNA 3 days after axotomy. beta(3) mRNA expression in ipsilateral DRGs increased significantly compared with contralateral DRGs 3 days after axotomy. In in situ hybridization histochemistry, beta(1) mRNA was predominantly expressed in medium- to large-size neurons, whereas beta(2) mRNA was expressed in small- to large-size neurons. There were no significant differences in beta(1) and beta(2) mRNA between contralateral and ipsilateral DRGs 3 days after axotomy. In contrast, beta(3) mRNA was mainly expressed in small neurons and occasionally in medium- to large-size neurons, and beta(3) mRNA expression in small c-type neurons in ipsilateral DRGs was increased significantly compared with contralateral DRGs. We examined beta(3) mRNA expression with one of alpha subunits, Na(v)1.3-ir, in DRG neurons after axotomy using the double labeling method. We found a high percentage of coexpression in injured DRG neurons: 83.6+/ 2.8% of neurons expressing beta(3) mRNA were labeled for Na(v)1.3-ir; 70.1+/-3.1% of Na(v)1.3-ir neurons expressed beta(3) mRNA. We also examined the expression of beta(3) mRNA in DRG neurons in the SNI model, a neuropathic pain model. We used activating transcription factor 3 to identify axotomized neurons, and found that beta(3) mRNA up-regulation occurred mainly in axotomized neurons in the neuropathic pain model. These data strongly suggest that beta(3) expression in injured DRG neurons following axotomy might be an important pathomechanism of post-nerve injury pain in primary sensory neurons. PMID- 14522004 TI - Differential antinociceptive effects induced by a selective cyclooxygenase-2 inhibitor (SC-236) on dorsal horn neurons and spinal withdrawal reflexes in anesthetized spinal rats. AB - The aim of present study was to examine the effect of a selective cyclooxygenase 2 (COX-2) inhibitor SC-236 (4 mg/kg) on the simultaneous responsiveness of spinal wide-dynamic range (WDR) neurons and single motor units (SMUs) from gastrocnemius soleus muscles to mechanical stimuli (pressure and pinch) and repeated suprathreshold (1.5xT, the intensity threshold) electrical stimuli with different frequencies (3 Hz, 20 Hz) under normal conditions and bee venom (BV, 0.2 mg/50 microl)-induced inflammation and central sensitization. During normal conditions, the responses of SMUs, but not WDR neurons, to mechanical and repeated electrical stimuli (3 Hz, wind-up) were depressed by systemic administration of SC-236 as well as its vehicle (100% dimethyl sulfoxide (DMSO)). The after-discharges of both the WDR neurons and the simultaneously recorded SMUs after electrical stimuli with 20 Hz were markedly depressed only by SC-236, indicating that the mechanisms underlying the generation of the C-fiber mediated late responses and the after-discharges may be different. The enhanced responsiveness of both WDR neurons and SMUs to mechanical pressure stimuli (allodynia) and pinch stimuli (hyperalgesia) in the BV experiments was apparently depressed by SC-236, but not its vehicle. For electrical stimulation, the enhanced late responses and after discharges, but not early responses, of both the WDR neurons and the simultaneously recorded SMUs were markedly depressed only by SC-236. This indicates that different central pharmacological mechanisms underlie the generation of these enhanced early, late responses, and after-discharges during BV-induced inflammation. The data suggest that the COX-2 inhibitor SC-236 apparently depress the activities of both spinal cord dorsal horn neuron and spinal withdrawal reflex during BV-induced sensitization, indicating that COX-2 plays an important role in the maintenance of central sensitization. PMID- 14522005 TI - Identification of two types of synaptic activity in the earthworm nervous system during locomotion. AB - In the ventral nervous system of the earthworm, a central pattern generator and motor neurons are activated during locomotion. We have previously reported that bath application of octopamine (OA) induces fictive locomotion in the earthworm, and the burst frequency of electrical activity from the first lateral nerves increases with OA concentration. However, there are no reports concerning locomotor neural networks in the earthworm. To identify neural networks involved in fictive locomotion, we optically monitored activity-dependent fluorescent staining in the earthworm ventral nerve cord (VNC) with a styryl dye, N-(3 triethylammoniumpropyl)-4-(4-(dibutylamino)styryl)pyridinium dibromide (FM1-43), and a confocal laser scanning microscope. OA induces FM1-43 fluorescence in a dose-dependent manner, with bright fluorescent spots of 3-10 microm in diameter observed to be localized around specified neurons in the segmental ganglion of the VNC. We compared OA dose-response curves for FM1-43 fluorescence with the bursting frequency for fictive locomotion, and found that two types of curves could be identified: one fluorescence response shows a similar dose-dependency to that of the burst frequency, while another response has a higher sensitivity to OA. From these results, we suggest that OA acts as one of the neuromodulators for the earthworm locomotion. This is the first attempt to record motor and inter neuronal activities simultaneously in a locomotor network in the earthworm. PMID- 14522006 TI - Longitudinal follow-up study of adenoviral vector-mediated gene transfer of dopamine D2 receptors in the striatum in young, middle-aged, and aged rats: a positron emission tomography study. AB - Overexpression of dopamine D(2) receptors by adenoviral vector-mediated gene transfer in the rat striatum was evaluated by positron emission tomography in vivo and by ex vivo autoradiography in 5-, 13-, and 24-month-old Fischer 344 rats. Each rat had hemilateral gene transfer of D(2) receptors mediated by adenoviral vectors (AdCMV.DopD(2)R) in the striatum with contralateral striatal injection of control vectors (AdCMV.LacZ). At day 2 or 3 after vector injection positron emission tomography or ex vivo autoradiography was performed. The binding potential of a radiolabeled D(2) receptors ligand, [11C]raclopride, was significantly higher in the D(2) receptors gene-transferred striatum than the control side in each age group at a similar degree. The binding potential in the AdCMV.DopD(2)R-injected striatum of 24-month-old rats was similar to that in the AdCMV.LacZ-injected striatum of 5-month-old rats (0.99+/-0.14 versus 0.91+/ 0.08). A significant age-associated decrease of the binding potential of [11C]raclopride was found in the control vector-injected side, and a significant increase of the binding potential in the adenoviral vector-injected side in all three age groups, suggesting no aging effect on the overexpression of D(2) receptors. A group of rats underwent follow-up assessment by positron emission tomography. The overexpression of D(2) receptors decreased with time in all three groups; however, the decrease rate of the D(2) receptors expression was significantly smaller in the 24-month-old group than in the 5-month-old group. We confirmed that the adenoviral vector-mediated gene transfer of D(2) receptors compensated the decreased density of striatal D(2) receptors in the 24-month-old rats up to the level in the control striatum of 5-month-old rats, and the decrease rate of the overexpression was significantly smaller in aged rats. PMID- 14522008 TI - Constant light housing attenuates circadian rhythms of mPer2 mRNA and mPER2 protein expression in the suprachiasmatic nucleus of mice. AB - Constant light (LL) or constant dark (DD) environmental lighting conditions cause a free-running period and activity reduction in the rodent behavioral circadian rhythm. In order to understand the molecular process underlying behavioral rhythms in LL or DD housing conditions, we examined the circadian profile of mPer2 mRNA and mPER2 in the suprachiasmatic nucleus (SCN), a main oscillator, of free-running mice. The circadian expression rhythm of mPer2 in the SCN was dampened under 7-day LL conditions, whereas that of mPER2 protein was moderately attenuated and its expression peak delayed. The circadian expression of mPer2 and its product was slightly attenuated and advanced by 7-day DD conditions. With arrhythmic behavioral activity caused by long-term LL housing, mPER2 protein lost its rhythmicity in the SCN. On the other hand, LL or DD housing did not affect the mPer2 gene and its product in the cerebral cortex. The present results suggest that mPER2 circadian expression in the SCN corresponds well with behavioral circadian oscillation under LL or DD conditions. Thus, the behavioral circadian rhythm seems to correlate with molecular clock works in the SCN. PMID- 14522007 TI - Efflux of human and mouse amyloid beta proteins 1-40 and 1-42 from brain: impairment in a mouse model of Alzheimer's disease. AB - Brain to blood transport is believed to be a major determinant of the amount of amyloid beta protein (AbetaP) found in brain. Impaired efflux has been suggested as a mechanism by which AbetaP can accumulate in the CNS and so lead to Alzheimer's disease (AD). To date, however, no study of the efflux of the form of AbetaP most relevant to AD, AbetaP1-42, has been conducted, even though a single amino acid substitution in AbetaP can greatly alter efflux. Here, we examined the efflux of AbetaP mouse1-42, mouse1-40, human1-42, and human1-40 in young CD-1, young senesence accelerated mouse (SAM) P8, and aged SAMP8 mice. The SAMP8 mouse with aging spontaneously overproduces AbetaP and develops cognitive impairments reversed by AbetaP-directed antibody or phosphorothioate antisense oligonucleotide. CD-1 mice transported all forms of AbetaP, although mouse1-42 and human1-40 were transported faster than the other forms. There was a decrease in the saturable transport of mouse1-42 in SAMP8 mice regardless of age. Efflux of mouse1-40 and human1-42 was only by a non-saturable mechanism in young SAMP8 mice and their efflux was totally absent in aged SAMP8 mice. These differences in the efflux of the various forms of AbetaP among the three groups of mice supports the hypothesis that impaired efflux is an important factor in the accumulation of AbetaP in the CNS. PMID- 14522009 TI - Immune rejection of a facial nerve xenograft does not prevent regeneration and the return of function: an experimental study. AB - Nerve grafts may be used to repair damaged peripheral nerves and also to facilitate spinal cord regeneration after experimental trauma. Little is known, however, about the possible use of xenografts and the role of immune rejection in the outcome of repair. In rats, excision of a short (7-8 mm) segment of facial nerve at its exit point from the skull base results in a permanent deficit in eye closure in the blink reflex. This deficit can be repaired by transplantation of a segment of either syngeneic rat facial nerve or xenogeneic Balb-C mouse sciatic nerve either with or without cyclosporine immunosuppression. With longer (15-20 mm) transplants, however, restoration of eye closure becomes dependent on cyclosporine administration. Thus, in a situation where nerve repair does not occur without a graft, a host immune attack has an attritional effect which is not sufficient to prevent repair over short distances, but becomes obvious when the regenerating fibres have to cross longer segments of transplanted tissue. PMID- 14522010 TI - Ampakines reduce methamphetamine-driven rotation and activate neocortex in a regionally selective fashion. AB - It has been proposed that glutamatergic and dopaminergic systems are functionally opposed in their regulation of striatal output. The present study tested the effects of drugs that enhance AMPA-receptor-mediated glutamatergic transmission (ampakines) for their effects on dopamine-related alterations in cortical activity and locomotor behavior. Rats with unilateral 6-hydroxydopamine lesions of the ascending nigro-striatal dopamine system were sensitized to methamphetamine and then tested for methamphetamine-induced circling behavior in the presence and absence of ampakines CX546 and CX614. Both ampakines produced rapid, dose-dependent reductions in circling that were evident within 15 min and sustained through 1 h of behavioral testing. In situ hybridization maps of c-fos mRNA expression showed that in the intact hemisphere, ampakine cotreatment markedly increased c-fos expression in parietal, sensori-motor neocortex above that found in rats treated with methamphetamine alone. Ampakine cotreatment did not augment c-fos expression in frontal, sensori-motor cortex or striatum. Still larger ampakine-elicited effects were obtained in parietal cortex of the dopamine depleted hemisphere where labeling densities were increased by approximately 60% above values found in methamphetamine-alone rats. With these effects, the hemispheric asymmetry of cortical activation was less pronounced in the ampakine cotreatment group as compared with the methamphetamine-alone group. These results indicate that positive modulation of AMPA-type glutamate receptors 1) can offset behavioral disturbances arising from sensitized dopamine receptors and 2) increases aggregate neuronal activity in a regionally selective manner that is probably dependent upon behavioral demands. PMID- 14522011 TI - Total sleep deprivation increases extracellular serotonin in the rat hippocampus. AB - Sleep deprivation exerts antidepressant effects after only one night of deprivation, demonstrating that a rapid antidepressant response is possible. In this report we tested the hypothesis that total sleep deprivation induces an increase in extracellular serotonin (5-HT) levels in the hippocampus, a structure that has been proposed repeatedly to play a role in the pathophysiology of depression. Sleep deprivation was performed using the disk-over-water method. Extracellular levels of 5-HT were determined in 3 h periods with microdialysis and measured by high performance liquid chromatography coupled with electrochemical detection. Sleep deprivation induced an increase in 5-HT levels during the sleep deprivation day. During an additional sleep recovery day, 5-HT remained elevated even though rats displayed normal amounts of sleep. Stimulus control rats, which had been allowed to sleep, did not experience a significant increased in 5-HT levels, though they were exposed to a stressful situation similar to slee-deprived rats. These results are consistent with a role of 5-HT in the antidepressant effects of sleep deprivation. PMID- 14522013 TI - Structure and characterization of hamster IL-12 p35 and p40. AB - Complementary DNAs coding for two subunits of hamster interleukin-12 (IL-12), p35 and p40, were cloned from a hamster dendritic cell (DC) cDNA library. The cloning demonstrated that hamster IL-12 consisted of a p35 subunit with 216 amino acid (aa) residues and a p40 subunit with 327 aa. Structural comparison of hamster p35 and p40 at the protein level showed the highest homologies with each counterpart of sigmodon (hispid cotton rat). The gene expressions of hamster IL-12 p35 and p40 in bone marrow (BM) cells cultured in the presence of mouse granulocyte macrophage-colony-stimulating factor (mGM-CSF) and IL-4 were up-regulated during culture. Immunoblot analysis of 293 cells transfected with hamster p35 and p40 expression vectors suggested the presence of a covalently linked p35/p40 heterodimer. Furthermore, supernatant from the 293 cells transfected with both expression vectors induced the up-regulation of interferon-gamma (IFN-gamma) mRNA in hamster splenocytes, indicating that the p35/p40 heterodimer IL-12 protein present in the supernatant was functional. These results suggest that the vectors containing hamster IL-12 cDNA might be suitable tools for developing an immunotherapeutic approach against experimental cancer in a hamster model. PMID- 14522012 TI - Structural analysis of IgG2A monoclonal antibodies in relation to complement deposition and renal immune complex deposition. AB - This study explores the structural features of murine monoclonal IgG2a anti dinitrophenyl (DNP) antibodies that were previously shown to form immune complexes (IC) differing in their capacity to bind complement, their clearance from the circulation and their deposition in the kidney. Interestingly, the sequence of one of these antibodies has a missing stretch of 14 amino acids within FR3. Molecular modeling suggests that this sequence deletion corresponds to the loss of beta-pleated sheet structure for two beta-strands (designated 4-3 and 4-4) on the external surface of the V(H) domain. Despite this sequence and conformational abnormality, the antibody retains affinity for DNP comparable to other IgG2a antibodies. Data presented here identify monoclonal IgG2a antibodies that form IC with varying propensity for both complement binding and renal deposition and yet have similar V(H) domain sequences. In fact, in the case of two IgG2a antibodies that form IC with very different renal tropisms and complement binding capacity, sequence variation within V(H) was observed only at three clustered residues within FR2, a single residue within FR3 and nine clustered residues spanning CDR3 and FR4. Sequence and modeling analysis also yielded the paradoxical finding that an antibody forming IC with a relatively high capacity to serve as a target for complement binding displays a relatively low number of solvent exposed acceptor residues for C4b and C3b. These data underscore the complex relationship between V domain structure, complement activation and renal deposition of model IC. PMID- 14522014 TI - Peptide immunogens based on the envelope region of HIV-1 are recognized by HIV/AIDS patient polyclonal antibodies and induce strong humoral immune responses in mice and rabbits. AB - Peptide immunogens produced by novel synthesis techniques and based on envelope proteins have repeatedly proven successful in inducing anti-HIV and anti-SIV responses in addition to producing anti-peptide immunity [Vaccine 12 (8) (1994) 736; AIDS Res. Hum. Retroviruses 14 (9) (1998) 751; Vaccine (2002) 2680]. We report here the design and evaluation of synthetic peptide constructs mimicking the variability of the third variable (V3) loop, or representing the conservation of the CD4 region of HIV-1 envelope. Three peptides based on the V3 region of HIV 1 subtype C gp120 and designated multiple epitope immunogens (MEI) differ in that one was randomly branched containing a spacer (b-MEI-s), another conjugated to a tetrameric lysine core (MEIV3b(4)) and the third conjugated to poly-L-lysine (poly-L-MEI). The method of synthesis employed produced peptides that were either linear, dimeric or tetramerically branched thereby providing differing levels of conformation. In addition the peptides also differed in their levels of variability. A fourth peptide designated B138 was based on a conserved region of HIV-1 envelope. The aim of this work was to evaluate the effect of variability and level of conformation on the immunogenicity of variable region peptides (MEI's) when compared to that of a known immunogenic and conserved region peptide (B138). Anti-peptide humoral immunity induced in BALB/c mice and New Zealand White rabbits produced antigen-recognizing antibody titres of 0.990). The limit of detection and the limit of quantitation were estimated as 0.7 and 2.0 microg/ml, respectively. Precisions in the range of 1.5-9.2 and 4.1-5.8% were obtained in intra- and inter-assay studies, respectively, using control samples containing 10, 14 and 26 microg/ml of caffeine. Accuracies ranging from 2.9 to 7.4% for intra-assay experiments, and from 3.9 to 5.4% in inter-assay studies were obtained. Stability of caffeine in urine samples was evaluated after long- and short-term storage at different temperature conditions. The batches of spiked urine were submitted to sterilization by filtration. No adsorption of the analyte on filters was observed. Before starting stability studies, batches of reference materials were tested for homogeneity. For long term stability testing, caffeine concentration in freeze-dried urine stored at 4 degrees C and in liquid urine samples stored at 4, -20, -40 and -80 degrees C was determined at several time intervals for 18 months. For short-term stability testing, caffeine concentration was evaluated in liquid urine stored at 37 degrees C for 7 days. The effect of repeated freezing (at -20 degrees C) and thawing was also studied for up to three cycles. The stability of caffeine was also evaluated in non-sterile samples stored at -20 degrees C for 18 months. No significant loss of the compound was observed at any of the investigated conditions. PMID- 14522022 TI - Sensitive liquid chromatography assay with ultraviolet detection for a new phosphodiesterase V inhibitor, DA-8159, in human plasma and urine. AB - A sensitive high-performance liquid chromatographic (HPLC) method with ultraviolet absorption detection (292 nm) was developed and validated for the determination of the new phosphodiesterase V inhibitor, DA-8159 (DA), in human plasma and urine. A single step liquid-liquid extraction procedure using ethyl ether was performed to recover DA and the internal standard (sildenafil citrate) from 1.0 ml of biological matrices combined with 200 microl of 0.1M sodium carbonate buffer. A Capcell Pak C18 UG120 column (150 mm x 4.6 mm I.D., 5 microm) was used as a stationary phase and the mobile phase consisted of 30% acetonitrile and 70% 20mM potassium phosphate buffer (pH 4.5) at a flow rate of 1.0 ml/min. The lower limit for quantification was 5 ng/ml for plasma and 10 ng/ml for urine samples. Within- and between-run accuracy and precision were < or =15 and < or =10%, respectively, in both plasma and urine samples. The recovery of DA from human plasma and urine was greater than 70%. Separate stability studies showed that DA is stable under the conditions of analysis. This validated assay was used for the pharmacokinetic analysis of DA during a phase I, rising dose study. PMID- 14522023 TI - Semi-automatic high-throughput determination of plasma protein binding using a 96 well plate filtrate assembly and fast liquid chromatography-tandem mass spectrometry. AB - A semi-automatic, high-throughput method has been developed to rapidly assess plasma protein binding of new chemical entities in drug discovery phase. New chemical entities are mixed with plasma and the unbound fractions are separated from the bound fraction by ultrafiltration in a 96-well filtrate assembly. The unbound fractions are then analyzed by fast liquid chromatography-tandem mass spectrometry (LC-MS/MS). Sample handling is automated by a robotic system. Employing a cocktail approach where multiple new chemical entities are allowed to bind to plasma proteins in the same well has further increased the throughput. We have validated the method with 12 commercially available compounds. The plasma protein binding data obtained by this method are comparable with the literature values. This method enables the determination of protein binding for 32 compounds in one single experiment instead of 1-2 compounds using the conventional methods. PMID- 14522024 TI - Human biomonitoring of pyrethrum and pyrethroid insecticides used indoors: determination of the metabolites E-cis/trans-chrysanthemumdicarboxylic acid in human urine by gas chromatography-mass spectrometry with negative chemical ionization. AB - This work describes a gas chromatographic-mass spectrometric method employing negative chemical ionization (NCI) for the determination of E-cis/trans chrysanthemumdicarboxylic acid (CDCA) in human urine used as a biomarker for the exposure to pyrethrum and/or certain pyrethroids in insecticide formulations applied indoors. Mixed-mode solid phase extraction was utilized for sample cleanup. Extraction recoveries ranged from 92 to 104% (2-9% R.S.D.). The acids were esterified with 1,1,1,3,3,3-hexafluoroisopropanol (HFIP) allowing both their gas chromatographic separation and their sensitive mass spectrometric detection under NCI conditions. Detection limits of ca. 0.05 microg/l urine were achieved. PMID- 14522025 TI - Simple determination of cyclosporine in human whole blood by high-performance liquid chromatography. AB - A simple and reproducible high-performance liquid chromatography (HPLC) method was developed for determination of cyclosporine (CyA, also known as cyclosporin A) in human whole blood. The method entailed direct injection of the blood samples after deproteination using acetonitrile. Chromatography was carried out using an ODS column under isocratic elution with acetonitrile-5mM disodium hydrogen phosphate (75:25, v/v), pH 5.1 at 70 degrees C and a detector set at 210 nm. The mean absolute recovery of cyclosporine from blood was 97%, and the linearity was assessed in the range of 100-3000 ng/ml blood, with a correlation coefficient of greater than 0.999. The limit of quantification and detection of the present method were 100 and 50 ng/ml, respectively. This method has been used to analyze several hundred human blood samples for bioavailability studies. PMID- 14522027 TI - Simple and efficient method for enantioselective determination of omeprazole in human plasma. AB - A practical and selective HPLC method for the separation and quantification of omeprazole enantiomers in human plasma is presented. C18 solid phase extraction (SPE) cartridges were used to extract the enantiomers from plasma samples and the chiral separation was carried out on a Chiralpak AD column protected with a CN guard column, using ethanol:hexane (70:30) as the mobile phase, at a flow rate of 0.5 ml/min. The detection was carried out at 302 nm. The method proved to be linear in the range of 10-1000 ng/ml for each enantiomer, with a quantification limit of 5 ng/ml. Precision and accuracy, demonstrated by within-day and between day assays, were lower than 10%. PMID- 14522026 TI - Quantitative determination of pioglitazone in human serum by direct-injection high-performance liquid chromatography mass spectrometry and its application to a bioequivalence study. AB - A simple, high throughput, direct-injection high-performance liquid chromatography tandem mass spectrometry method (LC/MS/MS) has been developed and validated for the quantitation of pioglitazone in human serum. After mixing the internal standard with a sample, a 10 microl portion of the mixture was directly injected into a high-flow LC/MS/MS system, which included an extraction column, an analytical column and a six-port switching valve. The on-line extraction was achieved on an Oasis HLB column (1 mm x 50 mm, 30 microm) with a 100% aqueous loading mobile phase containing 5 mM ammonium acetate (pH 4.0) at a flow rate of 4 ml/min. The extracted analyte was eluted by a mobile phase which contained 5 mM ammonium acetate and acetonitrile. The analytical column was a Luna C18 column (4.6 mm x 50 mm, 5 microm). Detection was achieved by positive ion electrospray tandem mass spectrometry. The lower limit of quantitation of the method was 9 ng/ml. The standard curve, which ranged from 9 to 1350 ng/ml, was fitted by a weighted (1/x2) quadratic regression model. The validation results demonstrated that this method had satisfactory precision and accuracy across the calibration range. There was no evidence of instability of the analyte in human serum following three freeze-thaw cycles, and samples could be stored for at least 2 weeks at -30 degrees C. This method was used to analyze pioglitazone concentrations in human serum samples from a bioequivalence study of a blinded Actos formulation (encapsulated 15 mg tablet) and an Actos 15 mg tablet. The blinded formulation was shown to be bioequivalent to an Actos 15 mg tablet. PMID- 14522029 TI - Feasibility of an on-line restricted access material/liquid chromatography/tandem mass spectrometry method in the rapid and sensitive determination of organophosphorus triesters in human blood plasma. AB - A rapid on-line solid phase extraction/liquid chromatography/tandem mass spectrometry (SPE/LC/MS/MS) method using restricted access material (RAM) was developed for the simultaneous determination of eight organophosphorus triesters in untreated human blood plasma. In a process involving column-switching techniques, the analytes were enriched on the RAM column, separated using a C-18 analytical column and detected with LC/MS. Tandem mass spectrometry was used to characterize and quantify the analytes. To elucidate the fragmentation pathway of a number of the analytes, MS3 experiments using an ion trap mass spectrometer were performed. The matrix effects associated with using APCI and ESI interfaces were investigated. The recoveries obtained were in the range 60-92% (R.S.D.<6%), with estimated detection limits between 0.2 and 1.8 ng/ml of plasma, and the total analysis time was 27 min. PMID- 14522028 TI - Determination of anti-cancer drug actinomycin D in human plasma by liquid chromatography-mass spectrometry. AB - Actinomycin D is an anti-cancer drug commonly used in the treatment of paediatric malignancies such as Wilms' tumour, Ewing's sarcoma and rhabdomyosarcoma. Despite its long history of clinical use, little is known about the pharmacokinetics of actinomycin D in humans, largely due to problems in developing an analytical assay with the required sensitivity to measure relevant clinical concentrations. As actinomycin D treatment in children with cancer is associated with veno occlusive disease (VOD), and as the dose intensity of actinomycin D treatment has been defined as a significant risk factor for the development of this potentially life-threatening hepatic toxicity, pharmacokinetic studies of actinomycin D may be beneficial in optimizing treatment with this drug. In order to investigate this issue, we developed a sensitive liquid chromatography-mass spectrometry (LC MS) method for the determination of actinomycin D in human plasma samples. Extraction of analytical samples was carried out with acetonitrile and analysis performed on an API 2000 LC/MS/MS using an internal standard of 7 aminoactinomycin D. A limit of quantitation of 1.0 ng/ml was determined, allowing the reliable measurement of actinomycin D in plasma samples obtained from patients receiving this drug clinically. The method demonstrated good reproducibility, over the calibration curve range of 1.0-100 ng/ml, with intra- and inter-assay precision CVs of 2.7-11.3 and 2.3-7.8%, respectively. Accuracy data showed relative errors of 2.0-16.4 and 10.4-15.2% for intra-assay (n=10) and inter-assay (n=7) experiments, respectively. Initial results of actinomycin D pharmacokinetics in paediatric patients are shown. PMID- 14522030 TI - Simultaneous determination of four antipsychotic drugs in plasma by high performance liquid chromatography. Application to management of acute intoxications. AB - A specific reversed phase-high pressure liquid chromatography (RP-HPLC) method has been developed for the simultaneous determination of clozapine (CZP), loxapine (LXP), zuclopenthixol (ZPT) and flupenthixol (FPT) in plasma. These four antipsychotic drugs are frequently used for the treatment of schizophrenia and other neuropsychiatric diseases. Carpipramine, a dihydrodibenzazepine, was used as an internal standard (I.S.). A liquid-liquid procedure was used to extract the drugs from human plasma. The analysis was performed on a XTerra MS C18 column with UV detection. Calibration curves were linear in the range 50-1000 microg/l. The limit of quantification (LOQ) was 15 microg/l for clozapine and loxapine and 20 microg/l for zuclopenthixol and flupenthixol. The coefficient of variation (CV) for intra- and inter-day precision was 7.2% or less with accuracies within 10% for the three concentrations.This isocratic and rapid method (run time<10 min) is useful for the management of acute intoxication. PMID- 14522031 TI - Liquid chromatographic determination of pyronaridine in human plasma and oral dosage form. AB - A new procedure for the determination of pyronaridine in plasma by reversed-phase high performance liquid chromatography (HPLC) with UV detection at 278 nm is described. The method involves liquid-liquid extraction of the drug with diethyl ether following basification of the deproteinized plasma with alkaline phosphate buffer. Chromatographic separation was achieved using a microbore C-18 column and a mobile phase consisting of 0.1% aqueous trifluoroacetic acid (TFA)-acetonitrile (75:25% (v/v)), pH 2.2, at a flow rate of 0.07 ml/min. Papaverine was used as internal standard. The response was linear between 50 and 1500 ng/ml. The limit of quantitation (LOQ) after plasma extraction was 50 ng/ml, the intra- and inter day precision ranged from 2.5 to 13.8% (CV). The recovery of the drug from plasma and accuracy were >90%. Preliminary application of the method for monitoring pyronaridine in humans upon oral administration of the tablet demonstrated the principal usefulness of the assay for clinical trial studies. The method can also be used to analyze the compound in pharmaceutical formulations. PMID- 14522032 TI - Liquid chromatography-tandem mass spectrometric quantitative determination of the HIV protease inhibitor atazanavir (BMS-232632) in human peripheral blood mononuclear cells (PBMC): practical approaches to PBMC preparation and PBMC assay design for high-throughput analysis. AB - A selective, accurate, and reproducible LC/MS/MS assay was developed and validated for the determination of the HIV protease inhibitor atazanavir (BMS 232632) in human peripheral blood mononuclear cells (PBMC) samples. In addition to the details of the validated LC/MS/MS method, a practical procedure is described in great detail for the preparation of large supplies of control (blank) PBMC from units of blood (each unit of blood is about 500 ml) for making the calibration standards and quality control (QC) samples. The PBMC assay design, intended for high-throughput sample analysis, is also described in some detail in regards to the composition and concentration expressions of the calibration standards and QC samples, the lysing procedure of the PBMC samples, and the final analysis/quantitation procedure. The method involved automated solid-phase extraction (SPE) of atazanavir and a stable isotope analog internal standard (I.S.) using 3M Empore C2-SD 96-well plates. A portion of the reconstituted sample residue was injected onto a YMC Basic analytical column which was connected to a triple quad mass spectrometer for analyte determination by positive-ion electrospray in the selected reaction monitoring (SRM) mode. The standard curve, which ranged from 5 to 2500 fmol per one million cells (fmol/10(6) cells), was fitted to a quadratic regression model weighted by 1/concentration. The lower limit of quantitation (LLOQ) was 5 fmol/10(6) cells. The inter- and intra-run coefficients of variation (CV) for the assay were <9% and the accuracy was 94-104%. Atazanavir was stable in PBMC for at least 24h at room temperature and for at least 129 days at -15 degrees C. PMID- 14522033 TI - Liquid chromatographic determination of oxaliplatin in blood using post-column derivatization in a microwave field followed by photometric detection. AB - Oxaliplatin ([(1R,2R)-1,2-cyclohexanediamine-N,N']oxalato(2-)-O,O'-platinum) is the first platinum drug with significant activity for metastatic colon cancer. The analysis of oxaliplatin has previously almost exclusively been based on the determination of the platinum content in plasma or ultrafiltrate using flameless atomic absorption spectroscopy (FAAS) or inductively coupled plasma mass spectrometry (ICPMS). A new method for quantitative determination of the free fraction of the intact drug in blood ultrafiltrate is presented here. Blood was ultrafiltrated centripetally at 4 degrees C and the ultrafiltrate was analyzed by liquid chromatography. Oxaliplatin was separated on a Hypercarb column using a mobile phase of methanol/succinic acid buffer pH 7.0 (9/1, v/v). Post-column derivatization was performed by adding N,N-diethyldithiocarbamate in methanol and with microwave heating of a Teflon tubing. The derivative was quantified by photometric detection at 344 nm. The coefficient of variation of standard blood samples was 4.9 and 2.5% at 0.100 and 1.00 microg/ml, respectively. The limit of quantitation was 0.04 microg/ml. PMID- 14522034 TI - Measurement of anti-cancer agent methoxyamine in plasma by tandem mass spectrometry with on-line sample extraction. AB - In this work, we present the development and validation of a tandem mass spectrometry method for the quantitative determination of methoxyamine (CH3ONH2), a potential new chemotherapeutic agent, in human and mouse plasma. Methoxyamine together with the internal standard (I.S.) methoxyl-D3-amine was directly derivatized in plasma sample with a novel chemical agent 4-(N,N diethylamino)benzaldehyde. The product solution was injected into an on-line Oasis HLB extraction column (2.1 mm x 20 mm) for analyte extraction. After the elution of extractives, the derivatized analytes were monitored by the positive electrospray-ionization mass spectrometry (ESI-MS-MS). The structures of derivatized analytes were elucidated by fragmentation. Quantitation of plasma methoxyamine was carried out by the multiple reaction monitoring (MRM) mode. This method had a linear calibration range of 1.00-1000 ng/ml with a correlation coefficient of 0.9999 for methoxyamine in both human and mouse plasma. The limit of detection (LOD) and limit of quantification (LOQ) for methoxyamine in plasma were 0.150 and 0.500 ng/ml, respectively. It was demonstrated that the method had high recovery and accuracy (90.1-94.7 and 90.1-96.3%), as well as excellent intra and inter-assay precision (2.2 and 3.7%), at three concentration levels (5.00, 50.0, 500 ng/ml). This method has been used to analyze the plasma levels of methoxyamine in samples obtained from male CD1 mice after bolus intraperitoneal injection of 2, 5 and 20mg methoxyamine hydrochloride (CH3ONH2.HCl) per kilogram mouse. PMID- 14522035 TI - Determination of 9-nitrocamptothecin by precolumn derivatization and its metabolite 9-aminocamptothecin in a biological fluid using reversed-phase high performance liquid chromatography with fluorescence detection. AB - A novel insoluble topoisomerase I inhibitor, 9-nitrocamptothecin (9-NC), is in advanced stages of clinical development and has been used to treat a diverse array of tumor types, including breast, ovarian, pancreatic and haematological malignancies. We have established a sensitive high-performance liquid chromatography method using fluorescence detection for the quantitation of 9-NC. Non-fluorescent 9-NC is converted to fluorescent 9-aminocamptothecin (9-AC) via a one-step pre-column derivative reaction. The quantitative limit of 9-NC was 1 ng/ml and the method was reproducible with the respective intra- and inter-day variability falling below 5.0 and 9.0%. The determination of both 9-NC and its metabolite 9-AC in dog plasma was also achieved using the same chromatographic and detection conditions. In dog plasma, the quantitative limits of 9-AC and 9-NC were 0.25 and 1 ng/ml, respectively. The presence of 9-AC in the samples yielded no interference with the determination of 9-NC. However, individual matrices can affect the conversion efficiency of 9-NC, thus indicating that standard samples should be run for each matrix. PMID- 14522036 TI - Determination of aminoethylcysteine ketimine decarboxylated dimer in human plasma and cultured cells by high-performance liquid chromatography with electrochemical detection. AB - Aminoethylcysteine ketimine decarboxylated dimer (AECK-DD) is a natural compound with antioxidant properties of a new family of sulfur-containing amino acids. It has been detected in human urine and plasma, in mammalian cerebellum and, more recently, in dietary vegetables. In the present study, a simple, highly sensitive method using a high-performance liquid chromatography system with electrochemical detection (ECD) has been developed. The method showed excellent precision and accuracy. It has been found to be about 100-fold more sensitive than gas chromatographic method and 2000-fold more sensitive in respect to the liquid chromatography method with UV detection. The method showed the required features of specificity and sensitivity to detect aminoethylcysteine ketimine decarboxylated dimer in human plasma and in cultured cells after in vitro supplementation. PMID- 14522037 TI - Simultaneous determination of theophylline and dyphylline by micellar electrokinetic chromatography and application in drug formulations. AB - A simple micellar electrokinetic chromatography is described for well resolution of theophylline, dyphylline and caffeine. The separation was performed at 25 degrees C using a background electrolyte consisting of 10mM borate buffer at pH 9 and 40 mM sodium dodecyl sulfate (SDS) as running buffer. Under this condition, good separation with high efficiency and short analyses time required is achieved. Several parameters affecting the separation of the drugs were studied, including the pH and concentrations of the borate buffer and sodium dodecyl sulfate. Using caffeine as an internal standard (I.S.), the linear range of the method for the determination of theophylline and dyphylline was over 0.03-1 micromol ml(-1); the detection limit (signal-to-noise ratio 3; injection 0.3 psi, 3s) was 0.01 and 0.02 micromol ml(-1), respectively. PMID- 14522038 TI - Ultra-fast quantitative bioanalysis of a pharmaceutical compound using liquid chromatography-tandem mass spectrometry. AB - This paper describes the ultra-fast determination of a pharmaceutical compound using TurboIonSpray LC-MS-MS on an API 4000 mass spectrometer. Sample preparation consisted of plasma protein precipitation, centrifugation and dilution of the supernatant. The use of small analytical column dimensions (2.1 mm x 10 mm) and high eluent flow rates (up to 2.2 ml/min) in isocratic mode led to a retention time of 9s. A sample-to-sample cycle time of only 10s was achieved by coupling two autosamplers. Partial separation of the drug and its main metabolite could be obtained. The d5-labeled drug used as internal standard compensated for matrix suppression effects. The assay was linear in the concentration range 1-1000 ng/ml, using standards prepared in human plasma. Inter-assay accuracy and precision were 98.5 and 6.2%, respectively. Mean intra-assay accuracy and precision calculated from quality control (QC) samples in human, rat and dog plasma at 3, 30 and 800 ng/ml were 100.8 and 3.8%, respectively. The ultra-fast LC-MS-MS method was successfully cross-validated against a commonly used column switching LC-MS-MS assay with 2.3 min run time by analyzing real study samples. PMID- 14522039 TI - Determination of urinary S-phenylmercapturic acid by liquid chromatography-tandem mass spectrometry. AB - Urinary S-phenylmercapturic acid (S-PMA) is considered a useful biomarker for the measurement of low levels of benzene exposure, related to occupational exposure, smoking habits or environmental pollution. S-PMA quantitative analysis requires highly sensitive and specific techniques and purification procedures, mainly based on liquid-liquid or solid-phase extraction, which result in time expensive analyses. A method was developed for the quantitative determination of S-PMA in urine by using a simple, reproducible and easily automatizable HPLC purification followed by LC/ESI-NI/MS2 analysis. In order to reduce the cost of the analysis, related to the use of expensive labeled standards, p-bromo-S-phenylmercapturic acid (p-Br-S-PMA) was synthesized, characterized and used as internal standard. The feasibility and efficacy of the proposed method were examined by constructing calibration curves in the range from 6.2 to 200 microg/l and data were analyzed in terms of linearity and statistical parameters. The detection limit, related to the purification of 1 ml urine sample is 5 microg/l. The method was applied to the analysis of 12 urine samples from smoker subjects non-occupationally exposed to benzene. S-PMA urinary levels ranged from 13.6 to >200 microg/l, suggesting a high influence of life style in the S-PMA excretion. The proposed analytical method is suitable for the biological monitoring of both smoker and non-smoker workers, occupationally exposed to benzene. By processing at least 2 ml of urine samples, the method appears to be also useful for the evaluation of benzene uptake due to the environmental pollution. PMID- 14522040 TI - Analysis of 12 different pentacyclic triterpenic acids from frankincense in human plasma by high-performance liquid chromatography and photodiode array detection. AB - For the determination of pentacyclic triterpenes of the boswellic acid family in human plasma a novel sensitive method was developed combining serial extraction on diatomaceous earth and graphitized carbon black followed by reversed phase high-performance liquid chromatography (HPLC) and photodiode array detection. The overall average extraction yield of 12 different pentacyclic triterpenic acids was approximately 66%. The calibration graphs were linear with coefficients of correlation for all compounds greater than 0.999. The overall within-day and between-day coefficients of variation (CV) for the 12 pentacyclic triterpenic acids were 5.6 and 6.8%, respectively. This HPLC procedure delivers the analytical sensitivity, precision and accuracy required for clinical pharmacokinetic and therapeutic studies. PMID- 14522041 TI - Automated method for determination of glutardialdehyde residues in flexible endoscopes after disinfection. AB - Glutardialdehyde (GDA) is the most commonly used disinfectant for flexible endoscopes. After inappropriate rinsing of endoscopes residual GDA in the narrow endoscope channels may lead to toxic effects in patients. Common methods for determination of aldehydes in water involve derivatization with 2,4 dinitrophenylhydrazine (DNPH), liquid-liquid or solid-phase extraction and HPLC determination. Since derivatization and extraction is both time-consuming and labor-intensive only a small number of samples can be measured. Thus, we developed a fully automated method which includes a conventional HPLC system, a programmable autosampler, and UV detection. After GDA derivatization using DNPH the samples remain in the aqueous phase and no preconcentration of the analyte is necessary. The samples are automatically derivatized through the autosampler. While derivatization in one sample takes place the previous sample is injected and measured by HPLC. Our method is well suited for screening residual GDA in endoscopes as it is both time- and labor-saving. PMID- 14522042 TI - Simultaneous determination of 3'-azido-2',3'-dideoxyuridine and novel prodrugs in rat plasma by liquid chromatography. AB - 3'-Azido-2',3'-dideoxyuridine (AZDU) is a nucleoside analog structurally similar to zidovudine (AZT) with proven activity against human immunodeficiency virus (HIV). The purpose of this study was to develop and validate a high-performance liquid chromatographic (HPLC) method to quantitatively determine AZDU and its novel prodrugs in rat plasma simultaneously. A reversed-phase gradient elution HPLC method was developed to quantitate AZDU and its prodrugs, N3 pivaloyloxymethyl-3'-azido-2',3'-dideoxyuridine (I), 5'-pivaloyloxymethyl-3' azido-2',3'-dideoxyuridine (II), 5'-O-valinyl-3'-azido-2',3'-dideoxyuridine hydrochloride (III) and 5'-O-phenylalanyl-3'-azido-2',3'-dideoxyuridine hydrochloride (IV), in rat plasma. AZDU and its prodrugs were analyzed using an octadecyl silane column with a mobile phase consisting of 0.04 microM sodium acetate buffer, pH 5.0, and acetonitrile, running in a segmented gradient manner at a flow rate of 2 ml/min. Acetonitrile was increased from 10 to 50% during the first 8 min by 5% per min, followed by 10% per min until it reached 90% acetonitrile. 3'-Azido-2',3'-dideoxy-5-ethyluridine (CS-85) was used as an internal standard (25 microg/ml). Compounds were detected by UV absorption at 261 nm. Extraction recoveries for all compounds were greater than 80%. Retention times of AZDU, CS-85, prodrugs I, II, III and IV were 3.3, 5.2, 9.1, 8.8, 6.3 and 7.3 min, respectively. Calibration plots were linear over the range of 0.25-100 microg/ml for AZDU and prodrugs II, III, and IV and 0.5-100 microg/ml for prodrug I. The limit of quantitation was 0.25 microg/ml for prodrugs II, III and IV and 0.5 microg/ml for prodrug I. The intra- and inter-day variations were less than 10% and accuracies were greater than 90%. This method is rapid, sensitive and reproducible for the determination of AZDU and prodrugs in rat plasma. PMID- 14522043 TI - Sensitive liquid chromatographic assay for the simultaneous determination of 5 fluorouracil and its prodrug, tegafur, in beagle dog plasma. AB - A high-performance liquid chromatographic (HPLC) method has been developed for the simultaneous determination of 5-fluorouracil (5-FU) and its prodrug, tegafur (TF), in dog plasma. 5-FU, the internal standard, 5-bromouracil, and TF were separated on a C18 Spherisorb ODS2 column using isocratic elution with retention times of 4.4, 8.0 and 21.2 min, respectively. Detection by UV absorption at 260 nm gave a limit of quantitation of 4 microg/l for 5-FU in plasma. Calibration curves for 5-FU and TF were linear over the ranges of 4-160 microg/l and 0.48 19.2 mg/l, respectively. Intra- and inter-day precision over these concentration ranges were <10.9 and <13.6% for 5-FU and TF, respectively, with good accuracy for both compounds. The method was successfully applied to define plasma concentration-time curves of TF and 5-FU in dogs administered a single oral dose containing TF (100 mg) and uracil (224 mg). PMID- 14522044 TI - Determination of irinotecan (CPT-11) and SN-38 in human whole blood and red blood cells by liquid chromatography with fluorescence detection. AB - An analytical method was developed for the anticancer agent irinotecan (CPT-11) and its main metabolite SN-38 in human whole blood and in red blood cells (RBCs). Sample pretreatment involved deproteinization of whole blood or plasma-diluted RBCs isolated by MESED instruments, with a mixture of aqueous perchloric acid and methanol (1:1, v/v). Separation was carried out using isocratic elution on a Hypersil ODS stationary phase, with detection at excitation and emission wavelengths of 355 and 515 nm, respectively. The lower limit of quantitation (LLQ) in blood was established at 5.00 ng/ml for both compounds, with values for within-run precision (WRP) and between-run precision (BRP) of less than 10%. The method is currently being applied to investigate the blood distribution of CPT-11 and SN-38 in cancer patients. PMID- 14522046 TI - ImmunoGen Inc. Immunoconjugates in canacer therapy: if at first you don't succeed, try, try again. PMID- 14522045 TI - Simultaneous determination of 2-methoxyphenol, 2-methoxy-4-methylphenol, 2,6 dimethoxyphenol and 4'-hydroxy-3'-methoxyacetophenone in urine by capillary gas chromatography. AB - A method for the simultaneous determination of 2-methoxyphenol, 2-methoxy-4 methylphenol, 2,6-dimethoxyphenol and 4'-hydroxy-3'-methoxyacetophenone in urine has been described. The metabolites were analyzed after enzymatic hydrolysis and extraction on octyl (C8) cartridges by using gas chromatography with flame ionization detection and a 5/95% copolymer of diphenyl-poly(dimethylsiloxane) capillary column. Methoxyphenols were well separated within 12 min. Recovery was over 90% in the range from 0.5 to 20 microg/ml; the detection limit was varying in the range of 0.05-0.11 microg/ml. The relative standard deviations and the accuracy were in the range of 3.1-15.5 and 2.4-16.0%, respectively. PMID- 14522047 TI - Bacteriophage that display small molecules. AB - Technology has been developed to display small molecules on phage particles. This innovation enables the generation of libraries of phage-tagged compounds with novel properties that are well suited for in vivo assays. PMID- 14522048 TI - Ontology recapitulates physiology. AB - High-content information experiments in the post-genomic era hold the promise of deciphering age-old questions in biology and new ones in the biomedical arena. In response, researchers are devising computationally intensive and novel strategies to extract answers from multidimensional data sets. PMID- 14522049 TI - The process of structure-based drug design. AB - The field of structure-based drug design is a rapidly growing area in which many successes have occurred in recent years. The explosion of genomic, proteomic, and structural information has provided hundreds of new targets and opportunities for future drug lead discovery. This review summarizes the process of structure-based drug design and includes, primarily, the choice of a target, the evaluation of a structure of that target, the pivotal questions to consider in choosing a method for drug lead discovery, and evaluation of the drug leads. Key principles in the field of structure-based drug design will be illustrated through a case study that explores drug design for AmpC beta-lactamase. PMID- 14522050 TI - Substrate 2'-hydroxyl groups required for ribozyme-catalyzed polymerization. AB - A polymerase ribozyme has been generated that uses nucleoside triphosphates to elongate an RNA primer by the successive addition of nucleotides complementary to an RNA template. Its polymerization is accurate, with an average error rate less than 3%, and it is general in terms of the sequence and the length of the primer and template RNAs. To begin to understand how the substrate contacts contribute to this accurate and general activity, we investigated which primer and template 2'-hydroxyl groups are involved in substrate recognition. We identified eight positions where 2'-deoxy substitutions can influence polymerization kinetics. All eight are within five nucleotides of the primer 3' terminus. Some, but not all, of the 2'-deoxy effects appear to be sequence dependent. These results begin to build a picture of how the polymerase ribozyme recognizes its substrates. PMID- 14522051 TI - Site-specific glycosylation of an aglycosylated human IgG1-Fc antibody protein generates neoglycoproteins with enhanced function. AB - A range of well-defined IgG glycoforms was prepared by employing a combination of synthetic carbohydrate chemistry and genetic engineering. The key aspect of this methodology is the coupling of thioaldoses with cysteine-containing proteins to give disulfide-linked neoglycoproteins. This technology was applied to the synthesis of a series of synthetic N-glycan thioaldoses which were coupled to an aglycosylated IgG1-Fc fragment, engineered to have Cys-297 in place of glycan linked Asn (Deltah-Fc N297C). Analysis of the resulting Fc neoglycoproteins by mass spectrometry and trypsin digestion showed that the saccharides were site selectively incorporated at Cys-297 to full occupancy without affecting other Fc protein disulfides. The neoglycoproteins were tested for their ability to interact with human FcgammaRI by inhibiting superoxide production by gamma interferon-stimulated U937 cells. The neoglycoproteins displayed enhanced superoxide inhibition relative to aglycosylated Deltah-Fc N297C, where increased glycan size correlated positively with increased inhibition. PMID- 14522052 TI - DNA polymerase template interactions probed by degenerate isosteric nucleobase analogs. AB - The development of novel artificial nucleobases and detailed X-ray crystal structures for primer/template/DNA polymerase complexes provide opportunities to assess DNA-protein interactions that dictate specificity. Recent results have shown that base pair shape recognition in the context of DNA polymerase must be considered a significant component. The isosteric azole carboxamide nucleobases (compounds 1-5; ) differ only in the number and placement of nitrogen atoms within a common shape and therefore present unique electronic distributions that are shown to dictate the selectivity of template-directed nucleotide incorporation by DNA polymerases. The results demonstrate how nucleoside triphosphate substrate selection by DNA polymerase is a complex phenomenon involving electrostatic interactions in addition to hydrogen bonding and shape recognition. These azole nucleobase analogs offer unique molecular tools for probing nonbonded interactions dictating substrate selection and fidelity of DNA polymerases. PMID- 14522053 TI - How do DNA repair proteins locate potential base lesions? a chemical crosslinking method to investigate O6-alkylguanine-DNA alkyltransferases. AB - O(6)-alkylguanine-DNA alkyltransferases directly reverse the alkylation on the O(6) position of guanine in DNA. This group of proteins has been proposed to repair the damaged base in an extrahelical manner; however, the detailed mechanism is not understood. Here we applied a chemical disulfide crosslinking method to probe the damage-searching mechanism of two O(6)-alkylguanine-DNA alkyltransferases, the Escherichia coli C-Ada and the human AGT. Crosslinking reactions with different efficiency occur between the reactive Cys residues of both proteins and a modified cytosine bearing a thiol tether in various DNA probes. Our results indicate that it is not necessary for these proteins to actively flip out every base to find damage. Instead they can locate potential lesions by simply capturing a lesioned base that is transiently extrahelical or sensing the unstable nature of a damaged base pair. PMID- 14522054 TI - Discovery of inhibitors that elucidate the role of UCH-L1 activity in the H1299 lung cancer cell line. AB - Neuronal ubiquitin C-terminal hydrolase (UCH-L1) has been linked to Parkinson's disease (PD), the progression of certain nonneuronal tumors, and neuropathic pain. Certain lung tumor-derived cell lines express UCH-L1 but it is not expressed in normal lung tissue, suggesting that this enzyme plays a role in tumor progression, either as a trigger or as a response. Small-molecule inhibitors of UCH-L1 would be helpful in distinguishing between these scenarios. By utilizing high-throughput screening (HTS) to find inhibitors and traditional medicinal chemistry to optimize their affinity and specificity, we have identified a class of isatin O-acyl oximes that selectively inhibit UCH-L1 as compared to its systemic isoform, UCH-L3. Three representatives of this class (30, 50, 51) have IC(50) values of 0.80-0.94 micro M for UCH-L1 and 17-25 micro M for UCH-L3. The K(i) of 30 toward UCH-L1 is 0.40 micro M and inhibition is reversible, competitive, and active site directed. Two isatin oxime inhibitors increased proliferation of the H1299 lung tumor cell line but had no effect on a lung tumor line that does not express UCH-L1. Inhibition of UCH-L1 expression in the H1299 cell line using RNAi had a similar proproliferative effect, suggesting that the UCH-L1 enzymatic activity is antiproliferative and that UCH-L1 expression may be a response to tumor growth. The molecular mechanism of this response remains to be determined. PMID- 14522055 TI - Synthetic compound libraries displayed on the surface of encoded bacteriophage. AB - We describe a technology for attaching libraries of synthetic compounds to coat proteins of bacteriophage particles such that the identity of the chemical structure is encoded in the genome of the phage, analogous to peptides displayed on phage surfaces by conventional phage-display techniques. This format allows a library of synthetic compounds to be screened very efficiently as a single pool. Encoded phage serve as extremely robust reporters of the presence of each compound, providing exquisite sensitivity for identification of active compounds engaged in complex biological processes such as receptor-mediated endocytosis and transcytosis. To evaluate this approach, we constructed a library of 980 analogs of folic acid displayed on T7 phage, and demonstrated rapid identification of compounds that bind to folate receptor and direct endocytosis of associated phage particles into cells that express the targeted receptor. PMID- 14522056 TI - Accessibility of nuclear chromatin by DNA binding polyamides. AB - Pyrrole-imidazole polyamides bind DNA with affinities comparable to those of transcriptional regulatory proteins and inhibit the DNA binding activities of components of the transcription apparatus. If polyamides are to be useful for the regulation of gene expression in cell culture experiments, one pivotal issue is accessibility of specific sites in nuclear chromatin. We first determined the kinetics of uptake and subcellular distribution of polyamides in lymphoid and myeloid cells using fluorescent polyamide-bodipy conjugates and deconvolution microscopy. Then cells were incubated with a polyamide-chlorambucil conjugate, and the sites of specific DNA cleavage in the nuclear chromatin were assayed by ligation-mediated PCR. In addition, DNA microarray analysis revealed that two different polyamides generated distinct transcription profiles. Remarkably, the polyamides affected only a limited number of genes. PMID- 14522057 TI - Cloning and characterization of the gene cluster encoding arthrofactin synthetase from Pseudomonas sp. MIS38. AB - Arthrofactin is a potent cyclic lipopeptide-type biosurfactant produced by Pseudomonas sp. MIS38. In this work, an arthrofactin synthetase gene cluster (arf) spanning 38.7 kb was cloned and characterized. Three genes termed arfA, arfB, and arfC encode ArfA, ArfB, and ArfC, containing two, four, and five functional modules, respectively. Each module bears condensation, adenylation, and thiolation domains, like other nonribosomal peptide synthetases. However, unlike most of them, none of the 11 modules possess the epimerization domain responsible for the conversion of amino acid residues from L to D form. Possible L- and D-Leu adenylation domains specifically recognized only L-Leu. Moreover, two thioesterase domains are tandemly located at the C-terminal end of ArfC. These results suggest that ArfA, ArfB, and ArfC assemble to form a unique structure. Gene disruption of arfB impaired arthrofactin production, reduced swarming activity, and enhanced biofilm formation. PMID- 14522058 TI - Biological mechanism profiling using an annotated compound library. AB - We present a method for testing many biological mechanisms in cellular assays using an annotated library of 2036 small organic molecules. This annotated compound library represents a large-scale collection of compounds with diverse, experimentally confirmed biological mechanisms and effects. We found that this chemical library is (1) more structurally diverse than conventional, commercially available libraries, (2) enriched in active compounds in a tumor cell viability assay, and (3) capable of generating hypotheses regarding biological mechanisms underlying cellular processes. We elucidated biological mechanisms relevant to the antiproliferative activity of 85 compounds from this library that were selected using a high-throughput cell viability screen. We developed a novel automated scoring system for identifying statistically enriched mechanisms among such a subset of compounds. This scoring system can identify both previously known and potentially novel antiproliferative mechanisms. PMID- 14522059 TI - Influenza virus and CNS manifestations. AB - Neurological involvement during influenza infection has been described during epidemics and is often consistent with serious sequelae or death. An increasing incidence of influenza-associated encephalitis/encephalopathy has been reported in Japan, mainly in children. A variety of other clinical CNS manifestations, such as Reye's syndrome, acute necrotising encephalopathy (ANE), and myelitis as well as autoimmune conditions, such as Guillain-Barre's syndrome, may occur during the course of influenza infection. Virological diagnosis is essential and based on virus isolation, antigen detection, RNA detection by PCR, and serological analyses. Neuroimaging with CT and MRI of the brain are of prognostic value. The pathogenic mechanisms behind the influenza CNS complications are unknown. The treatment is symptomatic, with control of vital functions in the intensive care unit, antiepileptic medication and treatment against brain oedema. PMID- 14522060 TI - Early diagnosis of SARS coronavirus infection by real time RT-PCR. AB - BACKGROUND: A novel coronavirus was recently identified as the aetiological agent of Severe Acute Respiratory Syndrome (SARS). Molecular assays currently available for detection of SARS-coronavirus (SARS-CoV) have low sensitivity during the early stage of the illness. OBJECTIVE: To develop and evaluate a sensitive diagnostic test for SARS by optimizing the viral RNA extraction methods and by applying real-time quantitative RT-PCR technology. STUDY DESIGN: 50 nasopharyngeal aspirate (NPA) samples collected from days 1-3 of disease onset from SARS patients in whom SARS CoV infections was subsequently serologically confirmed and 30 negative control samples were studied. Samples were tested by: (1) our first generation conventional RT-PCR assay with a routine RNA extraction method (Lancet 361 (2003) 1319), (2) our first generation conventional RT-PCR assay with a modified RNA extraction method, (3) a real-time quantitative RT-PCR assay with a modified RNA extraction method. RESULTS: Of 50 NPA specimens collected during the first 3 days of illness, 11 (22%) were positive in our first generation RT-PCR assay. With a modification in the RNA extraction protocol, 22 (44%) samples were positive in the conventional RT-PCR assay. By combining the modified RNA extraction method and real-time quantitative PCR technology, 40 (80%) of these samples were positive in the real-time RT-PCR assay. No positive signal was observed in the negative controls. CONCLUSION: By optimizing RNA extraction methods and applying quantitative real time RT-PCR technologies, the sensitivity of tests for early diagnosis of SARS can be greatly enhanced. PMID- 14522061 TI - SARS-coronavirus replicates in mononuclear cells of peripheral blood (PBMCs) from SARS patients. AB - BACKGROUND: The etiologic agent of severe acute respiratory syndrome (SARS) is a recently identified, positive single-stranded RNA (ssRNA) coronavirus (SARS-CoV). Little is known about the dynamic changes of the viral replicative form in SARS cases. OBJECTIVES: Evaluate whether SARS-CoV can infect and replicate in peripheral blood mononuclear cells (PBMCs) of infected persons and reveal any dynamic changes to the virus during the course of the disease. STUDY DESIGN: Peripheral blood mononuclear cells collected from SARS cases infected by the same infectious source were tested for both negative-stranded RNA (minus-RNA, "replicative intermediates") and positive-stranded RNA (genomic RNA) of SARS-CoV during the course of hospitalization by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: SARS-CoV minus-RNA was detected in PBMCs from SARS patients. The viral replicative forms in PBMCs were detectable during a period of 6 days post-onset of the disease, while the plus-RNA were detectable for a longer period (8-12 days post-onset). CONCLUSIONS: SARS-coronavirus can infect and replicate within PBMCs of SARS patients, but viral replication in PBMCs seems subject to self-limitation. PMID- 14522062 TI - Severe Acute Respiratory Syndrome (SARS). PMID- 14522063 TI - Quality control measures for the serological diagnosis of hantavirus infections. AB - BACKGROUND: With society's rapidly increasing mobility, patients infected with severe viral infections can become seriously ill at any place in Europe and elsewhere. Improving the diagnostics of these infections is the most important step in detecting the pathogens and dealing with them, and for this purpose, quality control measures are essential tools. OBJECTIVES: To assess the diagnostic reality for rare hantavirus infections in Europe by (1) running a pre evaluation panel (four samples, sent out in 1999) to optimise sample preparation and shipping procedure and afterwards (2) starting an External Quality Assurance (EQA) program (20 samples, sent out in 2001). STUDY DESIGN: All samples sent out had to be tested for the presence of specific IgG and IgM antibodies against hantavirus. For the pre-evaluation panel, four samples were distributed (two samples IgG+/IgM-, one sample IgG-borderline/IgM-, one sample IgG-/IgM-), for the EQA 20 samples (six samples IgG+/IgM+, eight samples IgG+/IgM-, one sample IgG borderline/IgM-, five samples IgG-/IgM-). Thirteen laboratories took part in the pre-evaluation panel, 18 laboratories participated in the first EQA run. RESULTS: For the pre-evaluation panel, the participants reported correct results for 64% of the IgG-positive samples (85% excluding borderline-positive sample), and 92% for the IgG-negative sample. IgM testing was correctly negative in all laboratories. For the EQA, the participants reported correct results for 76% of the IgG-positive samples, and 97% correct results for the IgG-negative samples. For the IgM-positive samples, 53% correct results were reported, and 98% correct results for the IgM-negative samples. CONCLUSIONS: The results presented here prove the importance of quality measures also for viruses only rarely suspected, like hantavirus, and they clearly demonstrate the need for improvement of the existing test systems. PMID- 14522064 TI - A comparison of methods for detecting adenovirus type 8 keratoconjunctivitis during a nosocomial outbreak in a Neonatal Intensive Care Unit. AB - BACKGROUND: An outbreak of epidemic keratoconjunctivitis (EKC) due to adenovirus (Ad) type 8 and involving 14 members of the hospital staff and 33 neonates admitted to the Neonatal Intensive Care Unit of the local University Hospital occurred between September and December 2000 in Pavia, Italy. The outbreak was preceded by an outbreak of EKC within the community. OBJECTIVE: To compare the performance of conventional virus isolation on cell cultures, direct detection of Ad antigens in conjunctival cells by a direct fluorescent assay (DFA) and Ad DNA detection in conjunctival swabs by polymerase chain reaction (PCR) for diagnosis of adenoviral conjunctivitis. STUDY DESIGN: Of conjunctival swabs collected from 47 patients, all were tested by virus isolation, 43 by direct Ad antigen detection, and 37 by Ad DNA detection. Direct Ad antigen detection was carried out by DFA using a group-specific monoclonal antibody. Detection and subgrouping of Ad DNA by nested PCR was performed using two sets of primers complementary to hexon and fiber genes, respectively. RESULTS: Ad was detected in 24/47 (51.1%), 21/43 (48.8%), and 23/37 (62.1%) samples by virus isolation, direct antigen detection and PCR, respectively. Overall, 30/47 (63.8%) samples were Ad-positive. Of 37 specimens tested in parallel by all three methods, Ad was detected by at least one of the three techniques in 26/37 (70.3%). All Ad isolates were identified as serotype 8 by neutralization, while all PCR-positive samples were identified as belonging to subgroup D. No other virus was isolated from any conjunctival swab. Time required for test completion was 9.6 (4-20) days for virus isolation, 1-2 h for DFA and 24 h for PCR. CONCLUSIONS: DFA was a sensitive and rapid assay but results depend on the quality of sample and the expertise of the observer. PCR was the most sensitive assay, although it takes longer to perform and requires dedicated facilities; thus, it could be restricted to DFA negative samples. Virus isolation is still useful from an epidemiological point of view. PMID- 14522065 TI - Polyomavirus BK DNA quantification assay to evaluate viral load in renal transplant recipients. AB - BACKGROUND: Several studies have disclosed a correlation between polyomavirus BK (BKV) and interstitial nephritis in renal transplant recipients and its quantification in urine and serum is therefore required to assess the role of BKV infection in nephropathy. OBJECTIVE: This paper describes a urine and serum BKV DNA quantification protocol devised to evaluate the viral load. STUDY DESIGN: Screening of samples containing > or =10(3)/ml viral genome copies by a semi quantitative polymerase chain reaction (PCR) assay is followed by precise quantification of the samples containing a high number of viral genomes in a quantitative-competitive (QC)-PCR assay. Generation of the competitor construct relied on the different sizes of wild-type and competitor amplicons. RESULTS AND CONCLUSIONS: Screening by semi-quantitative PCR selects samples with a high number of viral genomes for use in the more labor-intensive and -expensive QC-PCR assay and thus provides a handy means for quantitative DNA analysis of large numbers of samples. The results obtained in BKV-DNA quantification in urine and serum samples from 51 renal transplant recipients (22 on treatment with tacrolimus (FK506) and 29 on cyclosporine A (Cy A)) are interesting: BKV-DNA findings (43.1%) in urine samples are in agreement with the BKV urinary shedding reported in literature (5-45%). With regard to immunosuppressive treatment, the percentage of activation of the infection (revealed by BKV-DNA detection in urine samples) in the two groups of therapy is similar (40.9% vs 44.8%). The observation that the viral load in urine is dissociated with that of serum suggests that both parameters should be investigated in evaluation of the pathogenetic role of BKV reactivation in renal transplant recipients. Moreover, our BKV-DNA quantification protocol could be used to monitor viral load in urine and serum samples from renal transplant recipients so as to detect those at risk of nephropathy and monitor their response to immunosuppression reduction therapy if it occurs. PMID- 14522066 TI - Lymphoproliferative disease after allogeneic stem cell transplantation--pre emptive diagnosis by quantification of Epstein-Barr virus DNA in serum. AB - BACKGROUND: Lymphoproliferative disease (PTLD) is a life-threatening complication of organ transplantation. In matched, allogeneic, non-T-cell-depleted stem-cell transplantations (SCT) the disease develops early but has been thought to be rare. OBJECTIVES: We determined by strict histopathological criteria the incidence of fatal Epstein-Barr-virus (EBV)-related PTLD in a large number of SCT, and assessed the diagnostic value of a real-time quantitative polymerase chain reaction (qPCR) for EBV-DNA in serum specimens. STUDY DESIGN: Of the 257 SCT performed in Helsinki during 1994-1999, 132 (51%) recipients were alive and 125 (49%) had succumbed by June 2001. The necropsies were analyzed for EBV related PTLD as evidenced by disseminated lymphocytic infiltrates labeled histochemically for antigens and RNA (EBER 1 and 2) detectable by in situ technology. From a subset of the PTLD cases (N=12) and a series of corresponding stem-cell recipient controls (N=36), consecutive samples of serum (N=103 and 364, respectively) were studied by qPCR for EBV-DNA, and the clinical data were reviewed. RESULTS: The post-mortem analysis revealed 18 cases of PTLD (14% of the deceased), all of whom had received intensive immunosuppressive treatment including anti-thymocyte globulin for treatment or prophylaxis of graft versus host disease (GVHD). By using qPCR all the PTLD patients became EBV-DNA positive, in progressively rising copy numbers. EBV-DNA was first detectable 70 (median; range 24-154) days after SCT or 23 (4-86) days before death; i.e. earlier than the symptoms which appeared 15 (2-85) days before death. Among the SCT controls, EBV-DNA occurred sporadically (in only 3.9% sera). CONCLUSIONS: qPCR for EBV-DNA in serum is a highly sensitive (100%) and specific (96%) diagnostic approach. Intensely immunosuppressed stem-cell recipients are at a great risk of developing PTLD, and should be carefully monitored for EBV-DNA, for pre-emptive treatment of this life-threatening disorder. PMID- 14522067 TI - Generation and characterization of a monoclonal antibody specific for human herpesvirus 6 variant A immediate-early 2 protein. AB - Two variants (A and B) of human herpesvirus 6 (HHV-6) can be isolated from humans, with each variant having unique biological properties. HHV-6 variant typing is mainly done following amplification of viral genomic DNA followed by restriction endonuclease digestion. Our objective was to generate a monoclonal antibody (mAb) that would allow us to discriminate between variants A and B of HHV-6. BALB/c mice were immunized with a recombinant glutathione-S-transferase protein fused to the immediate-early (IE) 2 protein from HHV-6 variant A. Following splenocytes fusion, one IgG1 kappa light chain mAb (P6H8) was isolated and found to react specifically with variant A IE2 protein in immunofluorescence and western blot assays. The P6H8 antibody represents a useful tool for both fundamental research and clinical applications allowing for the discrimination of infections caused by HHV-6 variants A or B. PMID- 14522068 TI - Co-detection and discrimination of six human herpesviruses by multiplex PCR ELAHA. AB - BACKGROUND: Herpesviruses are a significant cause of human morbidity. Traditional approaches to the identification of these viruses require infectious or at least antigenic virus. Multiplex PCR (mPCR) is capable of simultaneously amplifying a range of targets from a single preparation of nucleic acids and when combined with a suitable detection assay, it is capable of discriminating each of the amplicons. OBJECTIVES: Several methods have been described in the literature, however, they lack one or more significant design features required to suitably control a routinely applied nucleic acid amplification assay. We aimed to design a multiplex herpesvirus PCR that could co-amplify eight human herpesvirus targets plus an internal control (IC) molecule in a single tube. STUDY DESIGN: Primers were designed to target the DNA polymerase genes of each of the human herpesviruses. Synthetic controls were developed to act as templates for the evaluation of assay sensitivity and specificity and for development of an in house competitive quantitative PCR. Amplicon was discriminated using a simplified enzyme linked amplicon hybridisation assay (ELAHA). RESULTS AND CONCLUSIONS: For routine diagnostic use we reduced the number of herpesviral targets from 8 to 6 in order to maintain adequate clinical sensitivity. The ELAHA proved more sensitive than agarose gel electrophoresis. Additionally, 36 cytomegalovirus positive patients were examined with an in-house quantitative PCR-ELAHA which was developed to confirm that that the mPCR's co-detection limit of 10(2) copy of synthetic template per millilitre was relevant for use in detecting virus from clinical samples. The mPCR-ELAHA was then applied to the screening of 174 patient specimens resulting in a specificity of 98% and a sensitivity of 93%. This preliminary study demonstrated that the mPCR-ELAHA was a complete approach to the detection of herpesviruses from a range of clinical samples and disease states. PMID- 14522069 TI - Rapid detection and quantification of cell free cytomegalovirus by a high-speed centrifugation-based microculture assay: comparison to longitudinally analyzed viral DNA load and pp67 late transcript during lactation. AB - BACKGROUND: Human cytomegalovirus (HCMV) is reactivated in nearly every seropositive breastfeeding mother during lactation [Lancet 357 (2001) 513]. Conventional tissue culture (TC) and low-speed centrifugation-enhanced microtiter culture methods are not able to detect HCMV from milk during all stages of lactation. OBJECTIVES: Development of a sensitive and quantitative microculture technique to describe the dynamics of HCMV reactivation in different milk compartments during lactation. STUDY DESIGN: Milk samples were collected longitudinally from seropositive breastfeeding mothers of preterm infants. Native milk samples were separated into fraction 1 (aqueous extract of milk fat), fraction 2 (cell and fat free milk whey) and fraction 3 (milk cells). Each of these fractions was screened qualitatively (TC, nPCR, pp67 late mRNA) and quantitatively (high-speed centrifugation-based microculture, quantitative PCR). RESULTS: Prior to low-speed centrifugation-enhanced inoculation, virus particles were concentrated by high-speed centrifugation (60 min at 50,000 x g, 4 degrees C). Using fraction 2 we were able to describe the dynamics of viral reactivation during lactation. We present the course of the quantitative virolactia and DNAlactia and qualitative detection of HCMV pp67 late mRNA in milk whey of four mothers (three transmitters and one non-transmitter). In all these cases virolactia described an unimodal and self limited course. Peak levels of virolactia for transmitters (T1: day 44; T2: day 43; T3: day 50) were closely related the onset of viruria of the corresponding preterm infants (U1: day 39; U2a/U2b: day 44/57; U3: day 60). The courses of viral load coincidence with the courses of DNA load. CONCLUSIONS: We present a rapid and highly sensitive microculture method for the quantification of cell free HCMV from milk whey and aqueous extracts from milk fat. Viral reactivation during lactation describes an unimodal course. Our findings have strong implications for quality control of any virus inactivation procedure. PMID- 14522070 TI - A simplified cytomegalovirus pp65 antigenemia assay procedure. AB - A simplified cytomegalovirus (CMV) pp65 antigenemia assay using a one-step erythrocyte lysis, fixation and permeabilization process was compared with a standard protocol, the CMV CINAkit (Argene Biosoft) assay. The results were comparable, both quantitatively and qualitatively. The new method saves time. It also provides flexibility because the cell suspension can be stored so that test completion can be deferred if so desired. PMID- 14522071 TI - A semiquantitative PCR method (SQ-PCR) to measure Epstein-Barr virus (EBV) load: its application in transplant patients. AB - BACKGROUND: High Epstein-Barr virus load has been related to an increased risk of Posttransplant Lymphoproliferative Disorders (PTLD) in transplant recipients. OBJECTIVES: Development of a method to quantitate EBV DNA levels in peripheral blood mononuclear cells (PBMC) and evaluate its usefulness in transplant patients. STUDY DESIGN: We designed a semiquantitative nested PCR based on a limiting dilution analysis to detect high viral loads in PBMC. This method was applied to 25 healthy carriers, and 85 solid organ transplant recipients as follows: (A) 53 asymptomatic patients; (B) 24 symptomatic patients; (C) eight patients with PTLD. RESULTS: In healthy carriers the reciprocal of the limiting dilution (RLD) ranged between non-detected (ND) and 1, the median RLD was ND, which is equivalent to a viral load of <1 copy per 10(5) PBMC. In the transplant population the medians RLD (range) were: (A) asymptomatic group: ND (ND-64), median equivalent to a viral load of <1 copy per 10(5) PBMC; (B) symptomatic group: 4 (ND-256), median equivalent to a range of viral load of 4-64 copies per 10(5) PBMC. (C) PTLD group: 256 (16-16384), median equivalent to a range of viral load of 256-4096 copies per 10(5) PBMC. Statistically significant differences were found between all groups: A+B vs. C (P<0.0001); A vs. B (P<0.0001); A vs. C (P<0.0001), B vs. C (P<0.0001). We also observed a good correlation between viral loads and clinical findings in four follow-up patients. Considering the RLD=256 as a cutoff point to detect transplant patients with PTLD, resulted in sensitivity 75%, specificity 96.7%, positive predictive value 60%, negative predictive value 98.3%. CONCLUSION: This SQ-PCR method enables us to differentiate between transplant patients with and without PTLD; therefore, it could be applied as a marker for early detection of this pathology. PMID- 14522072 TI - Rotavirus detection and characterisation in outbreaks of gastroenteritis in aged care facilities. AB - BACKGROUND: Although rotavirus is a major cause of gastroenteritis in children, its role in adult gastroenteritis and the sensitivity of different methods for its detection in specimens collected from adults are less well understood. OBJECTIVES: (1) To examine the frequency and seasonality of rotavirus-associated gastroenteritis outbreaks in aged-care facilities in Victoria, Australia. (2) To determine rotavirus type in these outbreaks. (3) To determine whether other enteropathogenic agents are present in specimens from these outbreaks. (4) To examine the sensitivity of different methods (electron microscopy (EM), reverse transcription-polymerase chain reaction (RT-PCR), enzyme immunoassay (EIA) and latex agglutination (LA)) for the detection of rotavirus in specimens from adults. STUDY DESIGN: Specimens from gastroenteritis outbreaks in aged-care facilities forwarded to this laboratory for the years 1997-2000 were tested for enteropathogenic agents by various methods. Epidemiological, clinical and seasonal data from the rotavirus-positive outbreaks were analysed. RESULTS: Rotavirus was detected by EM in 18 out of 29 individuals associated with seven out of 53 (13%) gastroenteritis outbreaks in aged-care facilities; norovirus was detected in 22 outbreaks (42%) and astrovirus in one outbreak (2%). No mixed viral infection was found in any outbreak. All rotaviruses were typed as Group A by RT-PCR. The rotaviruses in the seven outbreaks were G-typed as follows: G2 (three outbreaks), G4 (two outbreaks), G1 (one outbreak) and G9 (one outbreak). The rotavirus-associated outbreaks were concentrated in mid-winter to mid-spring. The relative sensitivities of the Group A rotavirus detection methods (for the 29 specimens tested) were EM (18), first-round RT-PCR (11), second-round PCR (19), EIA-visual (19), EIA-photometric (19) and LA (13). CONCLUSIONS: In Victoria, Australia, outbreaks of gastroenteritis associated with rotavirus are quite common in aged-care facilities. They involve Group A rotavirus and have a winter/spring seasonality. G-types G1, G2, G4 and G9 were all detected. EIA, second-round PCR and EM proved sensitive methods for rotavirus detection whereas first-round RT-PCR and LA did not. PMID- 14522073 TI - A rapid hemi-nested PCR for HTLV-I detection. AB - A hemi-nested PCR approach was adopted to detect HTLV-1 infection in clinical samples of peripheral blood mononuclear cells (PBMCs) from subjects with positive or indeterminate serological results. Our results showed that the hemi-nested PCR quickly solved the diagnostic query, detecting the presence of proviral HTLV-1 DNA in two of the 252 patients with inconclusive serological results. The main advantage of this method are the typology of DNA extraction, allowing a consistent DNA recovery without amplification problems, the rapidity (4-5 hours), the performance of the assay and its comparable or better sensitivity than other HTLV-1 PCR formats. PMID- 14522074 TI - The cellular Pax-Hox-helix connection. AB - Basic helix-loop-helix (bHLH) transcription factors are important regulators of lineage determination during embryogenesis. Initial experiments in Drosophila showed that early neural selection and specification are dependent on atonal (ato) and members of the achaete-scute complex (as-c). In mammals, transcription factors homologous to as-c and ato are causally involved during development of organs throughout the body. Development of subsets of lineages in intestine, stomach, pancreas, lung, thyroid and placenta have been shown to be regulated by members of the as-c and ato families. These functional studies show that an individual bHLH transcription factor can regulate multiple developmental processes throughout the mammalian body, which implicates that extant as-c and ato transcription factors play a distinct function dependent on their cellular context. Based on the synergistic activation of the insulin, POMC and Pax4 promotors by bHLH and homeobox (Hox) protein complexes, we hypothesize that the underlying cellular function-modulating factors include members of the Hox and paired box (Pax) multigene families. These examples indicate that unique combinations of bHLH and Hox proteins, mediated by protein-protein interactions, might be responsible for activating cell-specific sets of target genes. PMID- 14522075 TI - Genomic structure and expression of the mouse ESET gene encoding an ERG associated histone methyltransferase with a SET domain. AB - ESET (ERG-associated protein with a SET domain, also called SETDB1) is a novel histone methyltransferase that catalyzes methylation of histone H3-lysine 9 (H3 K9). Here we describe the genomic structure and expression of the mouse ESET gene that gives rise to ESET protein and its alternative splicing product. ESET is a 36-kb single copy gene and full-length ESET transcript consisting of 22 exons. The splicing variant retains only the first 12 exons and thus lacks sequences encoding the methyl CpG-binding domain and the catalytic SET domain. The U2 type conserved GT/AG consensus sequence is present at all of the splicing junctions within the ESET gene. The transcription initiation site of the ESET gene was determined by 5'-RACE experiment and by primer extension. The 5'-flanking sequence of the ESET gene does not contain the consensus TATA box. Instead, this ESET promoter region has features such as SP1-binding sites that are typical of housekeeping genes. The ESET promoter was functionally active when tested in transfection and luciferase assay. Full-length ESET transcript appears to be ubiquitously expressed. While the SET domain-deficient splicing variant is present in immortalized cell lines, it is undetectable by RT-PCR in the majority of normal mouse tissues. PMID- 14522076 TI - GCF2: expression and molecular analysis of repression. AB - GC-binding factor 2 (GCF2) is a transcriptional repressor that decreases activity of the epidermal growth factor receptor (EGFR) and other genes. We have mapped the gene for GCF2 by fluorescence in situ hybridization (FISH) to chromosome 2q37. Sequence analysis of the GCF2 gene and cDNA showed that the gene consists of eight exons and introns and spans 73 kbp of DNA. Northern blot analysis showed that GCF2 mRNA was differentially expressed in many human tissues and cell lines. GCF2 mRNA was expressed as a 4.2 kb mRNA in most human tissues with the highest expression level in peripheral blood leukocytes and lowest expression in brain and testis. Additional transcripts of 6.6, 2.9 and 2.4 kb were found in some tissues but the only transcript detected in cancer cell lines was 4.2 kb with high levels found in seven Burkitts' lymphoma cell lines. Western blot analysis showed that GCF2 protein is present at high levels in Burkitts' lymphoma and several other cancer cell lines. GCF2 was found in both nuclear and cytoplasmic compartments in cells. Deletion mutants of GCF2 revealed that amino acids 429-528 are required for both DNA binding and repression of the EGFR promoter. Furthermore, GCF2 was able to substantially decrease activator protein 2 (AP2) enhancement of the EGFR promoter. Thus, GCF2 is a transcriptional repressor overexpressed in cancer cell lines with a role in regulating expression of the EGFR. PMID- 14522077 TI - Molecular characterization and expression of a divergent alpha-tubulin in planarian Schmidtea polychroa. AB - We report the cloning and sequencing of a cDNA from planarian Schmidtea polychroa (Platyhelminthes, Turbellaria, Tricladida) encoding for an unusual tubulin isoform (SpTub-1) which is specifically expressed in testis. Sequence comparison of SpTub-1 with other known tubulins reveals that it has the highest homology with alpha-tubulins, even though the analysis of the molecular features shows that this isoform is significantly divergent. Hybridization of SpTub-1 to restriction-digested genomic DNA to Southern blotting produced a multiple banding pattern indicating that in planarian, a tubulin multigene family exists. Using in situ hybridization, we showed that the transcript is specifically detectable in planarian testis, suggesting that it may play a role in spermatogenesis. PMID- 14522078 TI - Comparative expression of the human beta(2) and beta(3) adrenergic receptors in Saccharomyces cerevisiae. AB - The beta(3) adrenergic receptor (beta(3)AR) is the predominant beta subtype in human brown adipocytes and is essential for regulating thermogenic lipolysis. To establish a novel experimental system for the biochemical analysis of this protein, we engineered several yeast strains. We show that the sterol background of the host strain greatly modulates the beta(3)AR expression but not in the same way as it modulates the beta(2) adrenergic receptor (beta(2)AR), the other main studied adipocyte subtype. The human beta(3)AR expressed in yeast is N glycosylated but not phosphorylated. This latter characteristic distinguishes it from the beta(2)AR. We showed that both beta(2)AR and beta(3)AR follow the secretory pathway to the yeast plasma membrane (PM) and are degraded in the vacuole. In the yeast strains used in this work, the two receptors also share a common mechanism of direct signal transduction through the yeast G(alpha) protein, Gpa1p. These strains thus appear to be useful for biochemical and structural studies of the human beta(3)AR in an in vivo reconstitution system. PMID- 14522079 TI - Identification of a gene in Leishmania infantum encoding a protein that contains a SP-RING/MIZ zinc finger domain. AB - The SP-RING or Miz zinc finger domain that is related to the classical RING finger motif, defines a class of proteins that can act as E3-like factors in the pathway of small ubiquitin-related modifier (SUMO) conjugation. This family includes the mammalian protein inhibitor of activated STAT (PIAS) proteins and related proteins from lower eukaryotes. Here we report the existence of a gene in Leishmania infantum, present as two identical copies placed upstream of each MAT2 gene copy, and transcribed as a single approximately 2.2 kb mRNA both in the logarithmic and stationary phases of the promastigote stage. This gene encodes a 47 kDa protein that has been named LORIEN. LORIEN is circumscribed to the cell periphery and it is antigenic during L. infantum infection of dogs and hamsters. Strikingly, this novel protein contains a highly conserved SP-RING/Miz zinc finger domain, raising the possibility that a SUMO or ubiquitin-like system may exist in this microorganism. PMID- 14522080 TI - Identification and characterization of mouse MTO1 gene related to mitochondrial tRNA modification. AB - The nucleotide modification in tRNA plays a pivotal role in the fidelity of translational process. The defects in nucleotide modification have often been observed in the mutated mitochondrial tRNAs associated with human diseases. Recently, MTO1-like protein in bacteria and yeast has been implicated to be a component of tRNA modification pathway. Here we report the identification and characterization of mouse MTO1 homolog. The mouse MTO1 gene containing 12 exons encodes a 669-residue protein with a strong homology to the MTO1-like proteins of bacteria and yeast, related to tRNA modification. Functional conservation of this protein is supported by the observation that the isolated mouse MTO1 cDNA can complement the respiratory-deficient phenotype of yeast mto1 cells carrying P(R)(454) mutation. MTO1 is ubiquitously expressed in various tissues, but with markedly elevated expression in tissues of high metabolic rates. Furthermore, we showed that mouse Mto1 localizes in mitochondrion. These observations suggest that the mouse MTO1 is a structural and functional homolog of yeast MTO1, thereby playing a role in the mitochondrial tRNA modification and protein synthesis. PMID- 14522081 TI - Cloning and characterization of the promoter region of the bovine membrane tethering protein p115 gene and its regulation in mammary epithelial cells. AB - To elucidate the molecular basis of the expression of the membrane tethering protein p115, a genomic DNA clone including the 5'-flanking region, first exon, and part of the first intron of the p115 gene was isolated from a bovine genomic DNA library in lambdaFIX II. Nine transcriptional initiation sites (at -189, 183, -178, -177, -164, -151, -149, -133 and -129 from the translation initiation site) were determined by the CapSite Hunting method. The 5'-flanking region (a 2.6-kb fragment) of the bovine p115 gene ligated to a luciferase reporter gene displayed promoter activity in primary cultured bovine mammary epithelial (BME) cells and in human MCF-7 breast carcinoma cells. The luciferase reporter gene assays of the promoter deletion constructs suggested the region required for promoter activity of the bovine p115 gene. The region that retained promoter activity contained a potential nuclear respiratory factor-1 (NRF-1) binding site. In both the BME and MCF-7 cells, mutagenesis to impair the NRF-1 consensus sequence in the p115 promoter gave substantially lowered levels of luciferase expression. An electrophoretic mobility shift assay (EMSA) showed that the NRF-1 consensus sequence and nuclear protein formed a complex that was abolished by oligonucleotides containing the authentic NRF-1 binding site, and was also supershifted with an antibody to NRF-1. In luciferase reporter gene assays of the p115 promoter constructs, treatment of the MCF-7 cells with estradiol-17beta (E(2)), insulin, or both stimulated the p115 promoter activity correlating with the cell proliferation rate. These results indicate that the NRF-1 response element in the p115 promoter is important for promoter function, and that it involves the binding of NRF-1. Furthermore, the results suggested that p115 gene transcription is activated by E(2), by insulin, or by both in association with the stimulation of mammary epithelial cell proliferation. PMID- 14522082 TI - Bridging PNAs can bind preferentially to a deleted mitochondrial DNA template but replication by mitochondrial DNA polymerase gamma in vitro is not impaired. AB - Mutations in mitochondrial DNA (mtDNA) are an important cause of neurological and other human pathologies. In the vast majority of cases, supportive care only is available. Mutated and wild-type mtDNAs often coexist in the same cell. A strategy for treatment has been proposed whereby replication of mutated mtDNA is inhibited by selective hybridisation of a nucleic acid derivative, allowing propagation of the wild-type genome and correction of the associated respiratory chain defect. Peptide nucleic acid molecules (PNAs) can be designed to selectively target pathogenic mtDNA with single point mutations. Molecules harbouring deletions present a complex problem. Deletions often occur between two short repeat sequences (4-13 residues), one of which is retained in the deleted molecule. With the more common large repeats, it is therefore difficult to design an antigenomic molecule that will bind selectively under physiological conditions. Following limited success with antigenomic oligodeoxynucleotides (ODNs), we have repeated these studies with a series of bridging PNAs. Molecules complementary to the sequence flanking either side of the 13 bp 'common deletion' were synthesised. The PNAs demonstrated markedly greater affinity for the delete than to the wild-type template. In runoff assays using Klenow fragment, these PNAs selectively inhibited replication of the delete template. However, no selective inhibition was observed when a polymerase gamma-containing mitochondrial fraction was used. PMID- 14522083 TI - Identification and characterization of a Zea mays line carrying a transposon tagged ENOD40. AB - In Zea mays, two ENOD40 homologous were identified that show only 30% of sequence homology to each other. We identified line e40-mum1 carrying a Mu transposon inserted in ZmENOD40-1, the maize gene that has the highest homology to leguminous ENOD40. The insertion causes a dramatic reduction of the ZmENOD40-1 transcript level. Irrespective of this, homozygous e40-mum1 plants are still able to interact with mycorrhizal fungi. Furthermore, no phenotypic aberrations correlated to the presence of e40-mum1 have been identified and therefore it is suggested that Z. mays ENOD40 genes are functionally redundant despite their strikingly low homology. PMID- 14522084 TI - Nrf2, not the estrogen receptor, mediates catechol estrogen-induced activation of the antioxidant responsive element. AB - The antioxidant responsive element (ARE) plays an important role in the gene expression of phase II detoxification enzymes, such as NAD(P)H:quinone oxidoreductase 1 (NQO1), and NF-E2-related factor2 (Nrf2) is the transcription factor for the ARE-driven genes. Interestingly, estrogen receptor (ER) was reported to increase NQO1 gene expression through the ARE. In this study, we investigated the role of ER and Nrf2 in ARE activation using IMR-32 cells and mouse primary astrocytes. Among tested estrogen-related compounds, only catechol estrogens (i.e. 4-hydroxyestradiol) activated the ARE. Since 4-hydroxyestradiol induced ARE activation was not inhibited by either 17beta-estradiol or tamoxifen, and overexpression of ER-alpha decreased 4-hydroxyestradiol-induced ARE activation, ARE activation by catechol estrogen was independent of ER. Nrf2, however, was very important in the 4-hydroxyestradiol-induced ARE activation. 4 Hydroxyestradiol did not activate the ARE in Nrf2 knockout (-/-) primary astrocytes, but did activate the ARE when Nrf2 was transfected into Nrf2-/- astrocytes. In addition, dominant negative Nrf2 completely blocked 4 hydroxyestradiol-induced ARE activation in Nrf2+/+ astrocytes, and only 4 hydroxyestradiol induced Nrf2 nuclear translocation in IMR-32 cells. A selective phosphatidylinositol 3-kinase (PI3-kinase) inhibitor (LY294002) blocked 4 hydroxyestradiol-induced Nrf2 nuclear translocation and NQO1 activity induction in IMR-32 cells. Taken together, these observations suggest that 4 hydroxyestradiol activates the ARE by a PI3-kinase-Nrf2 dependent mechanism, not involving ER. PMID- 14522085 TI - Occurrence of two distinct types of tissue inhibitor of metalloproteinases-2 in teleost fish. AB - We have cloned for the first time two cDNAs encoding distinct types of tissue inhibitor of metalloproteinases-2 (TIMP-2) from teleost fish, Japanese flounder, and designated these types as jfTIMP-2a and jfTIMP-2b. The open reading frames of the jfTIMP-2a and jfTIMP-2b cDNAs are composed of 663 and 657 nucleotides and 221 and 219 amino acids, respectively. Both jfTIMP-2s contain 12 cysteine residues, which might form six disulfide bonds as in other animals' TIMP-2s. The predicted full-length amino acid sequence of jfTIMP-2a has lower identity to jfTIMP-2b (63%) than to those of human (74%) and chicken (73%) TIMP-2s, but higher than to those of other human TIMPs (TIMP-1: 39%, TIMP-3: 43%, TIMP-4: 45%), indicating that jfTIMP-2a is a common TIMP-2, while jfTIMP-2b is unique to Japanese flounder. However, the C-terminal region including the last three disulfide bonds of jfTIMP-2b has higher amino acid identity to those of other animal TIMP-2s than to that of jfTIMP-2a. Reverse-transcribed polymerase chain reaction (RT-PCR) analysis showed the mRNAs of jfTIMP-2a and jfTIMP-2b to be ubiquitously expressed in all tissues examined, but with different expression patterns. These findings suggest that the two distinct jfTIMP-2s might perform different functions in teleost tissues. PMID- 14522086 TI - Molecular characterization of a ribosome-associated Hsp70-homologous gene from Rhizopus nigricans. AB - A ribosome-associated Hsp70-homologous gene (Rnssb-1) was isolated from the genomic library of the filamentous zygomycete fungus Rhizopus nigricans. The nucleotide sequence of a genomic clone encoded the N-terminal part of a protein with high similarity to the yeast SSB ribosome-associated chaperones. The missing 3' end of the gene was obtained by 3' RACE. The Northern blot analysis showed that the Rnssb-1 gene is constitutively expressed and is not induced upon heat shock at 37 degrees C. The primary structure analyses revealed that the coding region of the Rnssb-1 gene is interrupted by at least four introns. Their splicing was not inhibited by exposure of the organism to heat shock as proven by RT-PCR. A Southern blot analysis of R. nigricans genomic DNA confirmed the presence of two additional gene copies of ribosome-associated Hsp70 genes in the fungal genome. PMID- 14522087 TI - Cloning and characterization of a full-length pronociceptin in zebrafish: evidence of the existence of two different nociceptin sequences in the same precursor. AB - We report the cloning and molecular characterization of a zebrafish pronociceptin gene, which is expressed in brain and intestine. This fish precursor codes for two different nociceptin peptides, one of which presents an opioid sequence in its N-terminus, being more similar to dynorphin A than its mammalian counterparts. Our results represent a new tool to investigate the function of nociceptin and its receptor in relation to pain and drug addiction. PMID- 14522088 TI - Differential distribution of NK cells in decidua basalis compared with decidua parietalis after uncomplicated human term pregnancy. AB - As pregnancy progresses, a characteristic decline in the percentage of CD56bright CD16- uterine natural killer (NK) cells occurs. Studies of term decidua, however, have focused only on leukocytes derived from decidua basalis, the site of implantation. The decidua parietalis, lining the remainder of the uterine cavity is another important region of the maternal-fetal interface that forms contact with fetal tissue at the end of the first trimester. The aim of this study was to evaluate possible differences in expression of CD16 and CD56 on leukocytes from normal term decidua basalis and decidua parietalis. Decidua basalis and parietalis samples were obtained from 30 placentas collected after elective cesarean section. Percentages of leukocyte subpopulations and NK cell subsets within the CD45+ cell fraction were determined by flow cytometry. In six decidual samples, concurrent immunohistochemical staining was performed. Higher percentages of CD56dim CD16+ NK cells and CD56- CD16+ cells were found in decidua basalis in comparison to decidua parietalis. In contrast, the percentage of CD56bright CD16- uterine NK cells was significantly higher in decidua parietalis. Immunohistochemical quantification supported flow cytometric results. We conclude that significant differences exist with respect to the distribution of NK cells in term decidua basalis and parietalis. Future functional studies may improve our understanding of their role at the maternal-fetal interface. PMID- 14522089 TI - Functional categorization of HLA-DRB1 alleles in rheumatoid arthritis: the protective effect. AB - Because of past recombination event, human leukocyte antigen (HLA) alleles that are not closely related in overall sequence may come to resemble each other in areas coding for peptide binding regions (PBR) of HLA molecules. Peptide binding is likely to be important for the role of HLA molecules in autoimmune disease. As a result, it has been suggested that a strategy of searching for HLA disease associations that groups alleles in functional categories based on PBR motifs may be more successful than conventional strategies based on studying different alleles. Using such functional categorization, we examined the possibility of discriminating subcategories of HLA-DRB1 alleles associated with rheumatoid arthritis (RA) in a Southern French population. HLA-DRB1 genotyping was performed by polymerase chain reaction with sequence-specific oligonucleotide hybridization or sequence-specific primers. HLA-DRB1 alleles were classified according to a functional categorization that defined seven similar subregion structures or restrictive supertype patterns (RSPs) within pocket 4 of HLA-DR peptide binding groove as the molecular basis for grouping these alleles. HLA-DRB1* RSPs "A," "De," "Q," "Dr," "E," " R," and "a" association with susceptibility or resistance to disease was then studied in 200 RA patients versus 200 controls. DRB1* RSP "A" containing the shared epitope alleles (DRB1*0101, *0102, *0401, *0404, *0405, *0408, *1001, *1402; odds ratio [OR] = 4.35; pc < 0.001) had a predisposing effect, with double-dose effect as expected, OR 6.68 (pc < 0.001). Among the six remaining RSPs, two had significantly protective effect: DRB1* RSP "De" (DRB1*0103, *0402, *1102, *1103, *1301, *1302, *1304; OR = 0.33; p(c) < 0.001), and DRB1* RSP "Q" (DRB1*0701; OR = 0.40; pc < 0.001). One had non-significantly protective effect: DRB1* RSP "Dr" (DRB1*08, *1101, *1104, *1106, *12, *1303, *16; OR = 0.68; p < 0.05, pc = not significant [NS]). Three had neutral effect: HLA DRB1* RSPs "E" (DRB1*0403, *0407, *0901, *1401; OR = 0.71; p = NS), " R" (DRB1*0301, *0302; OR = 0.76; p = NS), and "a" (DRB1*1501, *1502; OR = 0.94; p = NS). The functional categorization allowed us to discriminate among the HLA-DRB1 alleles those that confer a predisposing effect, a neutral effect, and a protective effect in RA. PMID- 14522091 TI - High frequency of HLA-B44 allelic losses in human solid tumors. AB - Human leukocyte antigen (HLA) class I downregulation, a frequent phenomenon observed in a variety of human tumors, favors tumor immune escape from T lymphocyte recognition. However, it is not known whether a particular HLA class I allele is lost more frequently than others. To address this question we analyzed HLA class I expression in tumor tissues derived from 300 patients diagnosed as having breast, colorectal, or laryngeal carcinomas. Cryostatic tumor sections and a broad panel of anti-HLA class I monoclonal antibodies were used. We found that the HLA-B44 allele was lost more frequently than other HLA class I alleles, and that the difference was not related with changes in HLA-B44 allele frequencies between patients and controls. In addition, we observed that 35% of the HLA-B44 negative tumors presented HLA haplotype loss associated with loss of heterozygosity. These tests were performed on DNA samples obtained from microdissected tumor tissues. The results seem to indicate that HLA class I allelic losses are not randomly distributed during tumor development but that some HLA class I alleles, and HLA-B44 in particular, are more frequently downregulated and may play an important role in immune escape mechanisms. PMID- 14522090 TI - CTLA4 polymorphism in Spanish patients with systemic lupus erythematosus. AB - The cytotoxic T-lymphocyte antigen 4 (CTLA4, CD152) gene is a positional and functional candidate gene to susceptibility to systemic lupus erythematosus (SLE) because CTLA4 gene maps in the described SLE risk region 2q33 and CTLA4 molecule has an inhibitory effect on T-cell activation. Several polymorphisms have been described in CTLA4 gene, among them, a T/C change at position -1722, a C/T transition at position -319, and another A/G transition at position +49. The aim of this study was to investigate the possible association of these polymorphisms with the susceptibility to SLE in 276 Spanish autochthonous patients using a healthy control group composed of 194 ethnically matched volunteer bone marrow donors. Genotyping of these CTLA4 positions was performed in SLE patients and controls using a polymerase chain reaction amplification refractory mutation system. The genotypic frequencies were in Hardy-Weinberg equilibrium in all patients. No differences in the distribution of the genotype frequencies between patients and controls were found in any case. Our results from the Spanish autochthonous population differ from those found in the Korean population regarding the involvement of the polymorphism located at -1722 in the susceptibility to SLE. PMID- 14522092 TI - No evidence of type 1 diabetes susceptibility genes in the region centromeric of the HLA complex. AB - There is strong evidence that DQB1, DQA1, and DRB1 alleles are not the only contributors to the human leukocyte antigen (HLA) linked type 1 diabetes (T1D) predisposition. Although the HLA complex is much studied for disease association, little is known about the neighboring centromeric region. We have previously found suggestive association on DQ2-DR3 haplotypes for marker D6S291, located 3.6 Mb centromeric of HLA-DQB1. This region on human chromosome 6 is syntenic to a part of the region adjacent to the mouse major histocompatibility complex (MHC) on chromosome 17, which has been suggested to harbor a susceptibility gene in mouse (Idd16). To evaluate a possible role of the region centromeric of HLA-DQB1 in human T1D, we have scanned the region with nine microsatellite markers in 267 T1D families from five different populations. Our results indicate that the characteristic strong linkage disequilibrium in the HLA complex does not extend into this region. Furthermore, we did not detect any consistent T1D association for the markers analyzed in the study. In conclusion, our data argue against the presence of any strong genetic susceptibility factors for T1D in the region centromeric of the HLA complex. PMID- 14522093 TI - Association of the HLA-DR15/HLA-DQ6 haplotype with development of choroidal neovascular lesions in presumed ocular histoplasmosis syndrome. AB - Associations of human leukocyte antigen DR2 (HLA-DR2) and HLA-B7 with presumed ocular histoplasmosis syndrome (POHS) in the United States has been previously described. However, these associations were determined by means of low resolution, complement-dependent cytotoxicity assays for HLA-A, HLA-B, and HLA-DR molecules. To determine whether POHS is associated with other HLA alleles within the HLA-A, HLA-B, HLA-DR, and HLA-DQ loci, we performed a case control study of 34 patients diagnosed with macular choroidal neovascular membrane secondary to POHS and 45 healthy control individuals. Peripheral blood-derived DNA from the study patients was typed for HLA genes by means of sequence-specific primers that gave low-medium allele resolution. Significant associations were observed between HLA-B7 (X2 = 14.30, pc = 0.004, relative risk = 8.23), HLA-DR15 (X2 = 29.08, pc = 0.000001, relative risk = 27.50), and HLA-DQ6 (X2 = 23.09, pc = 0.00001, relative risk = 27.43) and POHS. Because there are strong linkage disequilibria between HLA-DR15 (a subtype of HLA-DR2) and HLA-B7 as well as HLA-DQ6, the significantly higher association of HLA-DR15 and HLA-DQ6 with POHS as compared to HLA-B7 suggests that the former alleles mediate susceptibility to the disease. In conclusion, there is a significant association between the HLA-DR15/HLA-DQ6 haplotype and development of choroidal neovascular lesions in POHS. PMID- 14522094 TI - Classic Pars Planitis: strong correlation of class II genes with gender and some clinical features in Mexican Mestizos. AB - The purpose of this study was the investigation of human leukocyte antigen (HLA) genes in Mexicans with classical Pars Planitis (CPP). Seventy-nine unrelated patients and 204 healthy controls were studied. HLA-A, -B, and -C typing was done on T cells isolated with immunomagnetic beads. HLA-DRB1, -DQA1, and -DQB1 loci were typed by polymerase chain reaction-sequence-specific oligonucleotide probes. The significance and strength of HLA associations were assessed. Stratification analyses were performed to analyze correlations between HLA alleles and clinical manifestations or gender. The mean age of CPP patients was 10 years old. The disease was recurrent (21.3%); 58% were males and 89.6% were bilaterally affected. A 3-year follow-up demonstrated no other associated disease. DRB1*0802 was significantly increased (odds ratio [OR] = 2.8, etiologic fraction [EF] = 18.96%). In females, HLA-B51 (OR = 9.8) was associated with nonsymmetrical onset and HLA-Cw1 (OR = 4.7) with symmetrical onset; DRB1*0802 was increased in males (OR = 3.9, p =5.0 E-05, EF = 38.3%) and contributed to their symmetrical onset (OR = 4.6, p =4.6 E-06, EF = 29.4%). Corneal peripheral endotheliopathy correlated with DQB1*0602 in females (OR = 17, EF = 47.1%). A susceptibility allele of Amerindian ancestry is responsible for juvenile CPP in Mexicans; HLA-B locus contributes to severity in females and DRB1*0802 in males. CPP should be classified as an heterogeneous illness taking into account ethnicity, and clinical and genetic characteristics. PMID- 14522095 TI - Stromal-cell derived factor-1 chemokine gene variant is associated with type 1 diabetes age at onset in Japanese population. AB - Stromal-cell derived factor-1 (SDF-1) is a powerful chemokine that upregulates T cell migration and activation. The gene for SDF-1 is located near type 1 diabetes susceptibility locus IDDM10, suggesting a contribution by SDF-1 to the induction of diabetes. Recently the role of SDF-1 gene polymorphism in the clinical presentation of type 1 diabetes in French population has been reported. To test the putative involvement of SDF-1 gene polymorphism in predisposition to or clinical heterogeneity of type 1 diabetes in Japanese population, we conducted the case-control study. The SDF1-3'A variant (801 G to A in the 3'-untranslated region) was determined by the polymerase chain reaction-restriction fragment length polymorphism technique in 184 patients with abrupt-onset type 1 diabetes and 106 healthy control subjects. No significant difference in allele and genotype frequencies of SDF1-3'A variant was found between type 1 diabetic patients and healthy controls. However, the SDF1-3'A variant was strongly associated with early-onset diabetes in a recessive model (AA versus AG + GG, p = 0.017). The mean age-at-onset in patients carrying SDF1-3'AA genotype was significantly younger than that in patients with SDF1-3' AG or GG genotype (p = 0.028). The frequencies of SDF1-3' A variant were significantly increased in HLA DR4/9 patients compared with non-DR4/9 patients (p = 0.008). These results suggest that the SDF-1 gene polymorphism is associated with the age-at-onset of type 1 diabetes in Japanese population. PMID- 14522096 TI - Polymorphisms of IL-1B, IL-1RN, IL-2, IL-4, IL-6, IL-10, and IFN-gamma genes in the Korean population. AB - Cytokines play a crucial role in regulating the immune and inflammatory responses. The collective influence of several cytokines can regulate immune responses as complex as those underlying allograft rejections or autoimmune diseases. Polymorphisms in the regulatory regions of the cytokine genes may influence their expression. Therefore, the polymorphisms of cytokine genes are potentially important as genetic predictors of the disease susceptibility or clinical outcome. In 311 unrelated healthy Korean individuals, we investigated the polymorphisms of cytokine genes (interleukin-1 [IL-1], IL-2, IL-4, IL-6, IL 10, and interferon-gamma [IFN-gamma]), which had been previously reported to be associated with a number of immune diseases, transplant complications, and direct or indirect influences on the level of expression and production. And we also compared the results to those published for other populations. The genotype distributions were consistent with the assumption of the Hardy-Weinberg equilibrium, with the exceptions of IL-1B +3954 and IL-6-174 polymorphisms. The polymorphisms examined in this study were almost similar to that observed in Asian populations. There were significant differences of the polymorphisms, except for IL-4 receptor alpha +1902, between Korean and other populations. Comparing the alleles associated with higher level of expression and production, IL-1B +3954*T, IL-2-330*G, and IL-4-590*T alleles were significantly higher, and IL-1RN*A2, IL-10-1082*G, and IFN-gamma*2 alleles were lower in Koreans than other populations. Especially in IL-6 promoter -174 polymorphism, we found only the G allele associated with higher plasma IL-6 levels. In haplotype analysis of IL-10 promoter polymorphisms, the GCC haplotype, associated with higher expression of IL-10, was significantly lower in Koreans. These results may be helpful for understanding transplant-related complications, immune or autoimmune diseases, and malignant diseases in the Korean population. PMID- 14522097 TI - Microarray and microfluidic methodology for genotyping cytokine gene polymorphisms. AB - Cytokine genetic polymorphisms are the subject of disease-association studies that require large-scale human genotyping. Polymerase chain reaction based custom microarrays and microfluidics systems were used to develop genotyping assays for following cytokine polymorphisms: tumor necrosis factor-alpha G-308A, interleukin 4 (IL-4) C-589T, interferon-gamma (CA)n repeats, IL-1RN 86-bp variable number of tandem repeats (VNTR), and CCR5 32-bp indel. For G-308A, 70.9% of DNA samples assayed were homozygous for wild type, 25.5% were heterozygous, and none were homozygous for variant allele. For C-589T, 35.5% of DNA samples were homozygous for wild type, 38% were heterozygous, and 22% were homozygous for variant. For IL 1RN VNTR, 71% of DNA samples were homozygous and the remainder were heterozygous. For CCR5, 96.4% of amplicons were homozygous for wild type, and 3.6% were heterozygous containing deletion. For IFN-gamma (CA)n repeats, 35.6% had 2,2 alleles, 42.2% had 2,3 alleles, and 11% had 3,3 alleles with alleles 1 through 5 corresponding to 11 through 15 repeats, respectively. There was good concordance between the results we obtained and current "gold-standard" methodologies for analyzing single nucleotide polymorphisms and size polymorphisms. Electronic DNA concentration with high stringency predisposes microarray technology to hybridization fidelity and accuracy, and microfluidics systems outperform conventional methodologies for size polymorphisms. Comprehensive genotyping can be achieved for clinical epidemiologic studies on cytokine gene polymorphisms using this approach. PMID- 14522098 TI - The role of probiotics in the treatment and prevention of Helicobacter pylori infection. AB - Literature reporting activity of probiotics in infections due to Helicobacter pylori has been reviewed to assess their value in combating such infections. Several in vitro studies show that lactobacilli or their cell-free cultures inhibit or kill H. pylori, prevent its adhesion to mammalian epithelial cells and prevent IL8 release. In vivo models demonstrate that pre-treatment with a probiotic can prevent H. pylori infections and/or that administration of probiotics markedly reduced an existing infection. Thirteen clinical trials have been published. In six (180 patients), a probiotic was used alone; five of these had an encouraging result-in three there were significantly reduced breath test readings and in two others some patients were cleared of infection. In seven further trials (682 patients), probiotics were added to a therapeutic regimen of antibiotics, resulting in an increased cure rate in two studies, and reduced side effects in four. Trials in which fermented milk products or whole cultures of lactobacilli were used tended to show better results than when the probiotic was taken in the form of bacteria alone. Not all the studies were randomised, double blind and placebo controlled, and some involved only small numbers of patients. The results suggest that probiotics may have a place as adjunctive treatment in H. pylori infections and possibly in prophylaxis. Future trials should address in particular the type of patient (asymptomatic volunteers, symptomatic patients), choice of probiotic strain(s), a wide range of probiotic strains (Lactobacillus acidophilus, L. johnsonii, L. gasseri, lactobacillus GG, Bifidobacterium longum, and bioyoghurts) have been used-some non-viable, regimens (doses and duration) and criteria of success (breath test, histology, culture, serology). PMID- 14522099 TI - Clinically significant borderline resistance of sequential clinical isolates of Klebsiella pneumoniae. AB - Two sequential clinical isolates of Klebsiella pneumoniae (Kpn) were isolated from bronchoalveolar lavage fluid (Kpn#1) and sputum (Kpn#2) of a patient with pneumonia, complicated by anatomical and immunosuppressive problems due to Wegener's granulomatosis. Despite 4 weeks of systemic treatment with ciprofloxacin (CIP) Kpn#2 was isolated thereafter. A fluoroquinolone-resistant mutant (Kpn#1-SEL) was derived from Kpn#1 in vitro by selecting on agar plates supplemented with ofloxacin. Kpn#1, Kpn#1-SEL and Kpn#2 had an identical pattern in PFGE. CIP MICs were 0.25, 2 and 4 mg/l for Kpn#1, Kpn#2 and Kpn#1-SEL, respectively. Kpn ATCC 10031 (CIP MIC 0.002 mg/l) served as control. We analyzed mechanisms of fluoroquinolone resistance by determining antibiotic susceptibility, organic solvent tolerance, accumulation of fluoroquinolones, dominance testing with wild-type topoisomerase genes (gyrA/B, parC/E), sequencing of the quinolone resistance determining regions of gyrA/B, parC/E and marR and Northern blotting of marR and acrAB genes. Compared with Kpn ATCC 10031, elevated MICs to fluoroquinolones and unrelated antibiotics in Kpn#1 was presumably due to a primary efflux pump other than AcrAB and increased the CIP MIC 125-fold. Although Kpn#1 tested sensitive according to NCCLS breakpoints, the elevated CIP MIC of 0.25 mg/l presumably rendered this isolate clinically resistant and lead to therapeutic failure in this case. Further increase of MIC to fluoroquinolones in vivo and in vitro was distinct. Kpn#1-SEL, selected in vitro, acquired a GyrA target mutation, whereas in Kpn#2 no known resistance mechanism could be detected. PMID- 14522100 TI - Simultaneous detection of mutations associated with resistance to macrolides and quinolones in Campylobacter jejuni and C. coli using a PCR-line probe assay. AB - Quinolone and macrolide resistance of Campylobacter jejuni and Campylobacter coli mainly depend on a mutation in gyrA and in 23S rDNA, respectively. In order to detect quinolone and/or macrolide resistant C. jejuni and C. coli strains, a macrolide and quinolone line probe assay (MQ-LiPA) was developed and 42 C. jejuni and C. coli strains were tested to evaluate the efficiency of the assay. Profiles of the mutations in 23S rDNA and in gyrA characterized by MQ-LiPA agreed with resistance to macrolides and quinolones. MQ-LiPA is a rapid and simple method for simultaneous detection of quinolone and macrolide resistance of C. jejuni and C. coli. We could also discriminate between C. jejuni and C. coli using probes for detection of gyrA mutations in MQ-LiPA. PMID- 14522101 TI - Characterisation of Leishmania donovani promastigotes resistant to hexadecylphosphocholine (miltefosine). AB - Leishmania donovani promastigote lines resistant to hexadecylphosphocholine (HePC, miltefosine) at 2.5, 5.0, 10.0, 20.0 and 40.0 microM were developed in vitro by continuous step-wise drug pressure. The 40 microM line was 15 times more resistant to HePC than the wild-type clone and showed cross-resistance to the ether lipid ET-18-OCH3 (edelfosine) but not to the standard anti-leishmanial drugs. Resistance was stable up to 12 weeks in drug-free culture medium. No amplification of specific genes, including the multidrug resistance P glycoprotein gene, could be detected in the resistant parasites. PMID- 14522102 TI - Mutation patterns of the reverse transcriptase genes in HIV-1 infected patients receiving combinations of nucleoside and non nucleoside inhibitors. AB - A genotyping assay was used to define human immunodeficiency virus type 1 (HIV-1) reverse transcriptase codons in plasma samples from 80 HIV-1 patients extensively treated with two nucleoside reverse transcriptase (zidovudine and lamivudine) and one non nucleoside reverse transcriptase (nevirapine) inhibitor. The frequencies of T215S/Y/F, M41L, D67N, L210W K70R, K219Q mutations, detectable in plasma samples, conferring resistance to zidovudine were 61.2, 56.2, 36.2, 31.5, 27.5 and 17.5%, respectively. Mutations (M184V or M184I) conferring resistance to lamivudine were detected in an extremely high percentage of patients (61%). Among mutations correlated to high (K103N, V106A, Y181C/I, Y188C/H/L, G190A/C/E/Q/S/T) or moderate (V108I, V118I) levels of nevirapine resistance, the predominant amino acid change was a substitution at 103 codon, present in 24 of 80 samples tested. Finally Q151M, the marker mutation able to confer resistance to all nucleoside analogues, was detected in seven patients with a viral load of between 1 x 10(4) and 9 x 10(4) HIV-1 RNA copies/ml. The relationship between the genotype and the viral load showed that the incidence of some specific mutations [M41L, T215Y (correlated to zidovudine resistance) and K103N (correlated to all NNRTIs drugs)] significantly (P=0.001) increased with higher viral load. Our results, albeit limited to a small cohort, showed a high frequency of mutations correlated to drugs in use, suggesting a need for therapeutic change in the near future and demonstrating that the development of genotyping tests helps to guide the therapeutic management of HIV-1 infected people. Our data highlight the dangers of selecting antiretroviral therapy without previous antiretroviral drug testing. Although the cost of these assays is a concern, prescribing inefficacious drugs could create serious problems for HIV-1 patients. PMID- 14522103 TI - Salmonella bloodstream infections: report from the SENTRY Antimicrobial Surveillance Program (1997-2001). AB - Salmonella spp. are significant bloodstream pathogens and are routinely monitored for antimicrobial resistance by the SENTRY Antimicrobial Surveillance Program. Six hundred and one bloodstream infection (BSI) isolates of Salmonella spp., collected over a 5-year period (1997-2001) were tested for their susceptibility against 20 antimicrobial agents, comparing year and geographical region. Salmonella enterica serotype Typhi was the most frequently identified 'species' (43% of identified strains), although 'unspeciated' strains predominated overall (54.2%). The rank order for six selected drugs tested by their MIC(90) values and percentage susceptibility was: ceftriaxone (< or =0.25 mg/l; 99.5% susceptible)>ciprofloxacin (0.12 mg/l; 99.3%)> trimethoprim/sulphamethoxazole (< or =0.5 mg/l; 92.7%)>amoxycillin/clavulanate (16 mg/l; 89.7%)>ampicillin (>16 mg/l; 81.0%)>tetracycline (>8 mg/l; 79.4%). Most isolates remained highly susceptible to all 20 agents examined, with the exception of Salmonella Typhimurium (only 35.3% susceptible to tetracycline, 41.2% to ampicillin, and 61.8% to amoxycillin/clavulanate). DT104 resistance phenotypes were noted in 3.4 and nearly 60.0% of unspeciated Salmonella and S. Typhimurium, respectively. Unexpectedly, the highest overall susceptibility rates were recorded in Latin America. Fluoroquinolone resistance was observed and nalidixic acid screening MICs (< or =8 mg/l) predicted full susceptibility to ciprofloxacin. Five-year results from the SENTRY Program show no clear trend toward greater resistances in Salmonella spp. BSIs for the commonly used antimicrobial classes. With the exception of S. Typhimurium DT104, most Salmonella spp. remain highly susceptible to the tested antimicrobials that maybe utilized for Salmonella BSI. PMID- 14522104 TI - Epidemiology and antibiotic susceptibility of bacteria causing skin and soft tissue infections in the USA and Europe: a guide to appropriate antimicrobial therapy. AB - Susceptibility data for all organisms associated with a range of skin and soft tissue infections (SSTI) in hospitalised patients were studied. Data were reported by clinical laboratories in the USA, France, Germany, Italy and Spain during 2001 which participate in The Surveillance Network (TSN). Staphylococcus aureus, Enterococcus spp. and coagulase-negative staphylococci (CNS), Escherichia coli and Pseudomonas aeruginosa were the most prevalent pathogens in all countries. MRSA was detected in 44.4, 34.7, 12.4, 41.8 and 32. 4% of S. aureus in each country, respectively. The majority of MRSA were cross resistant to other compound classes tested except for vancomycin (100% susceptible) trimethoprim sulphamethoxazole with range 1.7% (France) to 15.9% (Italy) resistant, and gentamicin with range 12.2% (France) to 87.0% (Italy) resistant. More than 99.0% of MSSA tested susceptible to ceftriaxone and >94.9% to trimethoprim sulphamethoxazole. 87.2% (France) to 94.6% of MSSA (Germany) were ciprofloxacin susceptible; 73.2% (USA) to 86.6% (Spain) were erythromycin susceptible; 85.4% (Italy) to 99.2% (France) were gentamicin susceptible. MSSA were more frequently found and generally more antibiotic susceptible from out patients. Overall, 100% of Streptococcus agalactiae and Streptococcus pyogenes were susceptible to penicillin, ceftriaxone and cefotaxime. Macrolide resistance was common among S. agalactiae (20.7%, Germany to 10%, Italy and Spain), S. pyogenes (19.2%, France to 11.1%, USA) and viridans streptococci (25.7%, France to 14.1%, Germany). Vancomycin-resistant Enterococcus spp. were uncommon outside the USA (17.5%) and Italy (7.4%). For all countries susceptibility of E. coli was 100% to imipenem, >98.7% to amikacin, >96.0% to ceftriaxone and cefotaxime. Susceptibility of E. coli isolates to ciprofloxacin was 77.6% in Spain to 94.3% in Germany. Klebsiella spp., Proteus spp., Citrobacter spp. and Enterobacter spp. displayed varying susceptibilities between countries to drugs tested. Putative extended spectrum beta-lactamase expression in E. coli remained rare comprising 4-5% of isolates in USA, Italy and Spain and in France and Germany <2%. For P. aeruginosa piperacillin-tazobactam, amikacin, imipenem and ceftazidime were the most active compounds tested irrespective of region. Surveillance data should be considered when selecting empirical therapy for treating SSTI. PMID- 14522105 TI - National antibiotic resistance monitoring in veterinary pathogens from sick food producing animals: the German programme and results from the 2001 pilot study. AB - In 2001, the first Germany comprehensive cross-sectional study into the sensitivity to antimicrobial substances of selected pathogenic bacteria from food producing animal species (dairy cows, fattening pigs) was conducted by the Federal Office of Consumer Protection and Food Safety, BVL (formerly, Federal Institute for Health Protection of Consumers and Veterinary Medicine, BgVV). Initial experience from national resistance monitoring revealed that the necessary organisational structures may be suitably established in a federal system. The quantitative sensitivity results (minimum inhibitory concentration, MIC) for the bacterial species examined showed lower resistance values in contrast to German data published previously and also in comparison with results from other European countries. Based on the experience from this pilot study, an urgent need has been identified to continue this interdisciplinary approach to tackle the resistance problem together with human medicine. After analysing the data from the pilot monitoring study, in 2002 the BVL started a year's study with an extended selection of bacterial species and indications. In the future, bacterial samples from private diagnostic laboratories and universities will also be included in the resistance monitoring system. PMID- 14522106 TI - A tissue cage model in calves for studies on pharmacokinetic/pharmacodynamic interactions of antimicrobials. AB - An in vivo model for studies of pharmacokinetic/pharmacodynamic (PK/PD) interactions of antimicrobials was developed. Tissue cages with a constant surface area but with different volumes were implanted in calves and infected with Mannheimia haemolytica. Penicillin was injected directly into the cages. With this procedure, different concentration-time profiles could be simulated so that the effect of a range of PK/PD indices on the infection could be monitored. The area under the curve to minimum inhibitory concentration (MIC) and time above MIC were equally predictive for effect, but Cmax to MIC was not. If drug dosages in relation to the MIC of strains used for infection are optimised, the model offers an interesting alternative to explore relevant factors for drug dosage optimisation. PMID- 14522107 TI - Increase of activity of tioconazole against resistant microorganisms by the addition of butylated hydroxyanisole. AB - Tioconazole (TCZ) killed resistant Candida albicans in less than 3 min, after the addition of butylated hydroxyanisole (BHA), a phenolic antioxidant, at subinhibitory concentrations. The bactericidal activity was also rapid against resistant Escherichia coli. BHA increased the TCZ activity in RPMI 1640 medium 18 times against ten strains of resistant C. albicans as judged by MIC and increased the activity 43 times against ten resistant E. coli strains. BHA at subinhibitory concentrations promoted the reduction of C. albicans virulence by reducing 180 times the hyphal cells of TCZ and decreasing hydrophobicity. The synergy could be due to changes in cellular permeability because of increased leakage of cellular enzymes. PMID- 14522108 TI - The susceptibility of non-fermentative Gram-negative bacilli to cefperazone and sulbactam compared with other antibacterial agents. AB - This study was designed to determine the bacterial susceptibility of non fermentative Gram-negative organisms to various antibacterial agents. Bacterial susceptibility testing used the Kirby-Bauer method and data were assessed according to NCCLS 2000. Cefoperazone/sulbactam (CPER/SU) had good antibacterial activity against Pseudomonas aeruginosa. Its activity was next only to that of imipenem, meropenem and ceftazidime. CPER/SU was highly active against Acinetobacter spp., Alcaligenes spp., Burkholderia spp., Stenotrophomonas maltophilia and Flavobacterium spp., while the majority of strains of the latter two species were resistant to imipenem and meropenem. Of 3905 isolates tested, 39.5% were susceptible to CPER, 70.4% to CPER/SU. The resistance rate was 37% for CPER and 10.8% for CPER/SU. PMID- 14522109 TI - AIDS-associated Cryptococcus infection before and after the highly active antiretroviral therapy era: emerging management problems. AB - The frequency, the microbiology and clinical features of AIDS-related primary episodes and relapses of cryptococcosis, before and after the introduction of highly active antiretroviral therapy (HAART), were compared. The study covered 58 cases diagnosed before the introduction of HAART, and eight episodes since 1997. Because of negative cultures, we sought a sensitive laboratory assay such as detection of polysaccharide antigen. Despite later diagnosis, there was reduced disease mortality. Clinical suspicion for HIV-associated cryptococcosis should be maintained in immunocompromised subjects. The introduction of HAART has led to significant clinical and laboratory changes of HIV-related cryptococcosis. PMID- 14522110 TI - Antibiotic susceptibility of strains isolated from patients with meningococcal disease and their household contacts in Catalonia (Spain). May 1998-April 1999. PMID- 14522111 TI - Drug resistance in Mycobacterium tuberculosis: a retrospective study from a 2000 bed teaching hospital in Ankara, Turkey. PMID- 14522112 TI - Clavispora (Candida) lusitaniae susceptibility profiles and genetic diversity in three tertiary hospitals (1998-2001). PMID- 14522113 TI - Lung-targeting microspheres of carboplatin. AB - Carboplatin (CPt) was incorporated in the gelatin microspheres by the method of emulsion and the drug content determined spectrophotometrically. The arithmetic mean diameter of the microspheres was 13.20 microm with 98% of the microspheres ranging from 5.0 to 28.6 microm. The average carboplatin content was 23.76% and the yield of the microspheres 85.12%. The microspheres were stable for three months when stored at 37 degrees C/RH 75%, showing insignificant change in appearance and drug content. The in vitro release profile of the microspheres could be described by a biexponential equation, and the release t(1/2) was 49.7 min and 92.04% released in 10h; while for the original drug, CPt, under the same conditions, 92.15% released in the first half an hour. Very high lung-targeting efficiency in vivo was proved by the results of targeting parameters. The S-180 lung neoplasm models were established by i.v. cancer cells in mice and the number of pulmonary nodules examined for evaluation of the treatment effect. The results of therapeutic tests showed that the antitumour effects were increased by injection of the microspheres compared with the injection of CPt solution: half of the dose in the microspheres showed comparable effect to the original drug. PMID- 14522114 TI - Preparation and in-vitro release rate of fentanyl-cyclodextrin complexes for prolonged action in epidural analgesia. AB - Fentanyl was complexed with cyclodextrin derivatives with the intention to obtain parenteral solutions able to provide prolonged analgesia following epidural administration. Three cylodextrins (CDs) suitable for parenteral use were used: hydroxypropyl-beta-cyclodextrin (HP-beta-CD), sulfobutylether-beta-cyclodextrin (SBE-7-beta-CD), and maltosyl-beta-cyclodextrin (malt-beta-CD). Analysis of fentanyl was done with HPLC-UV. The inclusion capacity of HP-beta-CD was determined from phase-solubility diagrams at pH 6.5, 7.2 and 8.0, and those of SBE-7-beta-CD and of malt-beta-CD at pH 8.0. Solubility of fentanyl increased linearly (i) as a function of the CD concentration, and (ii) with decreasing pH. Complexation was highest with HP-beta-CD and malt-beta-CD, much higher than with SBE-7-beta-CD, with stability constants at pH 8.0 of 801, 729 and 1309 M(-1), respectively. The CD concentration was calculated to obtain a fentanyl-CD formulation, with the desired amount free fentanyl as loading dose in solution and the rest complexed with CD, as reservoir for prolonged action. A suitable membrane and a release-rate apparatus were selected for in-vitro release-rate studies. Best results were obtained with Spectrapor membranes and a home-made release-rate apparatus. Release rate was evaluated in static and dynamic conditions. For both modes, the release rate of fentanyl decreased as a function of CD concentration, due to complex formation of fentanyl, which suggests the possibility to provide prolonged pharmacodynamic effects in vivo. PMID- 14522115 TI - Surface characterisation of bags for total parenteral nutrition by tensiometry and atomic force microscopy. AB - Bags made of poly-ethylene and poly-vinylchloride and of the copolymer ethylene vinylacetate were used as containers of perfusion solutions for total parenteral nutrition. The bags were characterised by tensiometry (free energy and its polar and dispersed components) and atomic force microscopy (AFM) before and after various periods of storage of solutions for total parenteral nutrition containing L-aminoacids, electrolytes or glucose. In most of the cases, after storage of these solutions, tensiometric characterisation and atomic force microscopy analysis of the internal surface of bags showed deep modifications which highlight the adsorption of the solutes. The changes of surface characteristics were found to depend on the time of contact, the wettability of the polymer and the compounds present into the solutions, while their concentration has a negligible effect. Generally, the aminoacid solutions produced a higher increase in the polar component even after short storage times. Poly-ethylene and the copolymer ethylene-vinylacetate showed a greater inertia if compared with the poly-vinylchloride bags. PMID- 14522116 TI - A novel delivery system for amphotericin B with lipid nano-sphere (LNS). AB - A low-dose therapeutic system with a lipid emulsion for amphotericin B (AmB), a potent antifungal drug, was studied. Lipid nano-sphere (LNS), a small-particle lipid emulsion, was taken up by the liver to a lesser extent than was a conventional lipid emulsion. As a result, LNS yielded higher plasma concentrations of a radiochemical tracer than did the conventional lipid emulsion. LNS was therefore judged to be a suitable carrier for a low-dose therapeutic system for AmB, and LNS incorporating AmB (LNS-AmB) was prepared. LNS AmB was found to be a homogeneous emulsion with mean particle diameters ranging from 25 to 50 nm. LNS-AmB yielded higher plasma concentrations of AmB than did Fungizone, a conventional intravenous dosage form of AmB, after intravenous administration to mice, rats, dogs, and monkeys. This difference between LNS-AmB and Fungizone was also observed for constant intravenous infusion. In contrast to Fungizone, LNS-AmB showed a linear relationship between dose and AUC. These pharmacokinetic characteristics of LNS-AmB make it a suitable candidate for an effective low-dose therapeutic system for AmB. PMID- 14522117 TI - Design and evaluation of an early stage drug release apparatus. AB - This paper describes the design and evaluation of an early stage drug release apparatus (ERA) to determine drug release from pellets at times less than 1 min. The apparatus comprises a stirred sample chamber in which the sample is retained by a BS150 mesh screen (0.106 mm), and a series of jacketed cups containing the medium (80 ml) which are raised and lowered in turn over the fixed sample chamber for specified periods. Three types of early release studies were used: single 60s study (single cup), 10s followed by 50s (two cups) and 10, 20, 30...60s multiple changeover differential release (six cups). The effects of stirrer speed, stirrer position and multiple changeover on drug release from standard paracetamol alginate pellets were investigated. Drug release rates from non-disintegrating pellets were reproducibly determined. The three types of early release study schemes yielded reproducible drug release data over sampling times less than 1 min. Stirrer speed, and depth, and changeover motion of release cups affected drug release but yielded reproducible results. Release from the standard pellets used to study the apparatus took 3 days to stabilize and remained stable thereafter. The apparatus can be used for screening of pellet formulations of sparingly soluble drugs during their developmental stage and regular quality assurance studies of pellets (>150 mesh size). Along with early release studies of pellets, it could be easily modified to study other types of formulations and for automation. PMID- 14522118 TI - A new index, the core erosion ratio, of compression-coated timed-release tablets predicts the bioavailability of acetaminophen. AB - Although compression-coated tablets are a commonly used timed-release drug delivery technology, their utility is often limited by poor bioavailability. To try to improve the bioavailability of these tablets, the effect of their core composition of compression-coated tablet on in vivo pharmacokinetics was investigated. First, the extent of mass reduction of cores in different compression-coated tablet core formulations was used to establish a new index, the core erosion ratio. The data show that adding excipients with high water solubility to the core results in a greater core erosion ratio. Next, to elucidate the effect of core erosion ratio on in vivo acetaminophen (AAP) release, three compression-coated tablet formulations with similar in vitro AAP release profiles but different core erosion ratios were administered to four fasted dogs. The time for first appearance (TFA) of AAP in plasma did not differ significantly among formulations, indicating that the in vivo lag time was the same for all formulations. In separate experiments, necroscopy revealed that 3h after oral administration, the tablets were located in the ileum and colon and that all three formulations had identical GI transit times. However, the area under the AAP plasma concentration-time curve was greater in dogs given formulations with larger core erosion ratios. These results suggest that a formulation with a large core erosion ratio can significantly increase in vivo drug release from compression-coated tablets, leading to increased drug absorption from the lower GI tract. PMID- 14522119 TI - Bioavailability of a morphine suppository is increased after intracolostomal administration in colostoma-constructed rabbits. AB - This study was performed to assess the pharmacokinetics of morphine and its major metabolites after its rectal or colostomal administration in rectal-resected (ROP) or colostoma-constructed (SOP) rabbits, respectively. The pharmacokinetics of morphine, morphine-3-glucuronide (M3G), and M6G in normal rabbits appeared to be similar to those in human, judging from their plasma concentration-time profiles and the susceptibility of morphine to first-pass metabolism. In SOP, but not ROP, rabbits, the plasma concentrations of morphine, M3G and M6G were significantly increased compared with those in normal rabbits. The AUC of morphine and its metabolites, and the F value of the former in the SOP group were greater than those in the control group, while the elimination half-life (t(1/2)) values were comparable in the two groups. In addition, the disposition of morphine and its metabolites after intravenous (i.v.) administration to SOP rabbits was almost the same as that in normal rabbits, suggesting that an increase in the rate of absorption of morphine in SOP rabbits was not due to inflammation at the absorption site caused by operation, but probably due to its increased solubility in loose stools. Therefore, great attention should be paid when morphine suppositories are intracolostomally administered to colostoma constructed patients. PMID- 14522120 TI - Effect of side-chain structures on gene transfer efficiency of biodegradable cationic polyphosphoesters. AB - Cationic polyphosphoesters (PPEs) with different side-chain charge groups were designed and synthesized as biodegradable gene carriers. Poly(N-methyl-2 aminoethyl propylene phosphate) (PPE-MEA), with a secondary amino group ( CH(2)CH(2)NHCH3) side chain released DNA in several hours at N/P (amino group of polymer to phosphate group of DNA) ratios from 0.5 to 5; whereas PPE-HA, bearing CH(2)(CH2)(4)CH(2)NH(2) groups in the side chain, did not release DNA at the same ratio range for 30 days. Hydrolytic degradation and DNA binding results suggested that side chain cleavage, besides the polymer degradation, was the predominant factor affected the DNA release and transfection efficiencies. The side chain of PPE-MEA was cleaved faster than that of PPE-HA, resulting poor cellular uptake and no transgene expression for PPE-MEA/DNA complexes in COS-7 cells at charge ratios from 4 to 12. In contrast, PPE-HA/DNA complexes were stable enough to be internalized by cells and effected gene transfection (3400 folds higher than background at a charge ratio of 12). Interestingly, gene expression levels mediated by PPE-MEA and PPE-HA in mouse muscle following intramuscular injection of complexes showed a reversed order: PPE-MEA/DNA complexes mediated a 1.5-2-fold higher luciferase expression in mouse muscle as compared with naked DNA injection, while PPE-HA/DNA complexes induced delayed and lowered luciferase expression than naked DNA. These results suggested that the side chain structure is a crucial factor determining the mechanism and kinetics of hydrolytic degradation of PPE carriers, which in turn influenced the kinetics of DNA release from PPE/DNA complexes and their transfection abilities in vitro and in vivo. PMID- 14522121 TI - Formation of fine drug particle by cogrinding with cyclodextrins. Part II. The influence of moisture condition during cogrinding process on fine particle formation. AB - The purpose of this study was to investigate the effect of moisture condition during cogrinding process on fine drug particle formation. Cogrinding of cyclodextrins (CDs) and pranlukast (PRK) hemihydrate was performed in various moisture conditions at a mixing molar ratio of 2:1 (CDs:PRK) and the formation of PRK submicron particle was investigated. The moisture content in the cogrinding process significantly affected the fine particle formation. More than 90% of pranlukast loaded transformed to submicron particles when coground with alpha-CD, beta-CD or gamma-CD containing the specific amount of water for each CD system. Fine particle formation of PRK was considered as a particular phenomenon to cyclodextrins, since the submicron particles could not be formed when D-mannitol, lactose or microcrystalline cellulose (MCC) was used as a cogrinding additive. Moreover, the appearance and disappearance of fine particle formation was found to be reversible depending on the existence of water during the grinding process. PMID- 14522122 TI - Improvement in the bioavailability of poorly absorbed glycyrrhizin via various non-vascular administration routes in rats. AB - The purpose of this study was to examine the improvement of the bioavailability of glycyrrhizin (GL) via extra-vascular, i.e. oral, rectal, and nasal routes with or without the aid of an absorption enhancer in place of the vascular intravenous route in rats. Pharmacokinetic behavior following administration via vascular routes, i.e. the intravenous and portal-venous routes was examined in rats. The area under the plasma concentration-time curve (AUC) after administration of GL via the portal vein was decreased slightly, suggesting that the first elimination of GL in the liver may be one of the factors contributing to the low bioavailability after administration via the oral route. When GL was administered orally as a solution (30 mg/kg), the plasma concentration of GL was extremely low. However, after rectal or nasal administration of GL solution (30 mg/kg) with or without sodium caprate, the mean AUC value was remarkably increased compared with oral administration. In particular, the absolute bioavailability of GL after nasal administration was estimated to be approximately 20%, which was approximately 80-fold greater compared with after oral administration despite of the absence of an enhancer. Furthermore, the fatty acids co-administered orally with GL produced an increase in GL absorption in the following order: sodium caprate>sodiumlaurate>sodiumcaprylate>sodium oleate. These results indicate that the rectum and nasal cavity are useful administration routes for systemic delivery of GL. It was also found that the fatty acids were enhancers for the absorption of GL. PMID- 14522123 TI - Ultrasonication of chitosan and chitosan nanoparticles. AB - The objective of this study was to evaluate the effects of ultrasonication on chitosan molecules and nanoparticles. Molecular weight (M(v)) of chitosan HCl (M(v) 146 kDa and degree of deacetylation (DD) 96%) decreased linearly with increasing duration and amplitude of ultrasonication. DD and FTIR absorption were unaffected. X-ray diffraction (XRD) analysis suggested greater chain alignment in the ultrasonicated chitosan samples. Chitosan nanoparticles had mean diameter of 382 nm, polydispersity of 0.53 and zeta potential of 47 mV. Ultrasonication administered at increasing duration or amplitude decreased the mean diameter and polydispersity of the nanoparticles. Zeta potential and FTIR absorbance were unaffected, while XRD suggested a greater disarray of chain alignment in the nanoparticle matrix. Under the transmission electron microscope (TEM), freshly prepared nanoparticles were dense spherical structures which became fragmented after ultrasonication for 10 min at amplitude of 80. Untreated nanoparticle formulation turned turbid upon storage for 3 weeks at ambient conditions due to substantial swelling of the nanoparticles. Ultrasonicated nanoparticle formulation remained clear on storage. Although the particles had also swelled, they were no longer spherical, assuming instead an irregular shape with branching arms. In conclusion, high-intensity ultrasonication induced considerable damage on the chitosan nanoparticles which could affect their function as drug carriers. PMID- 14522124 TI - Comparative evaluation of the human whole blood and human peripheral blood monocyte tests for pyrogens. AB - Two different in vitro tests for pyrogens, using human peripheral blood monocytes (PBMNC) and diluted whole blood (WBC), respectively, were applied to different classes of parenteral medicinal products. Many of these products did not have a specified endotoxin limit concentration that was established as the maximum valid dilution to comply with the test. The results of the in vitro tests for pyrogens were compared with the results from the Limulus amoebocyte lysate (LAL) and rabbit pyrogen tests. The Second International Standard for endotoxin was used to calibrate all of the assays and the International Standard for IL-6 was used to calibrate the IL-6 ELISA which provided the readout for the in vitro tests for pyrogens. Preparatory tests were conducted to ensure that the "criteria for validity and precision of the standard curve" were satisfied and that the drugs being tested did not interfere in the tests. The PBMNC/IL-6 test had a detection limit of 0.06 EU/ml and spike recoveries were 62-165%. The whole blood/IL-6 test also had a detection limit of 0.06 EU/ml and spike recoveries were 58-132%. The application to the detection of non-endotoxin pyrogens needs to be evaluated in more detail, but the two in vitro tests for pyrogens showed good agreement overall, both with each other and with the LAL test and the rabbit pyrogen test for the detection of endotoxins. PMID- 14522125 TI - Design and monitoring of photostability systems for amlodipine dosage forms. AB - Photostability of amlodipine (AML) has been monitored in several pharmaceutical inclusion systems characterized by plurimolecular aggregation of the drug and excipients with high molecular weight. Several formulations including cyclodextrins, liposomes and microspheres have been prepared and characterized. The photodegradation process has been monitored according to the conditions suggested by the ICH Guideline for photostability testing, by using a light cabinet equipped with a Xenon lamp and monitored by spectrophotometry. The formulations herein tested have been found to be able to considerably increase drug stability, when compared with usual pharmaceutical forms. The residual concentration detected in the inclusion complexes with cyclodextrins and liposomes was 90 and 77%, respectively, while a very good value of 97% was found for microspheres, after a radiant exposure of 11,340 kJm(-2). PMID- 14522126 TI - The physical stability of thermally-stressed phospholipid-based emulsions containing methyl, propyl and heptyl parabens as model drugs. AB - The objectives of the studies presented herein was to investigate the mechanisms of emulsion instability under thermal stress (121 degrees C) by evaluating the effects of a lipophilic drug dissolved in the internal phase of an oil-in-water (o/w) emulsion on growth rate suppression and the apparent microviscosity. Model drugs used were methyl, propyl and heptyl paraben. The o/w emulsions were prepared using medium chain triglycerides as an internal phase in aqueous glycerol solutions emulsified with phospholipids. Concentrations of paraben in the internal phase varied from 0.2-0.8 M. Microfluidization was used to reduce the droplet size to the submicron range. Microviscosity was calculated from the measured anisotropy of a fluorophore probe (1,6-phenyl-1,3,5-hexatriene) using a modified Perrin's equation. Emulsion aliquots were subjected to thermal stressed at 121 degrees C the droplet growth rate was determined from periodic measurements of the mean droplet diameter using photon correlation spectroscopy. The growth rate decreased in the presence of parabens. Maximal growth suppression occurred at paraben concentrations of 0.4 M. However in deference to theoretical predictions of the effects of increasing co-solute concentrations based on Ostwalt ripening, the droplet growth rates increased at concentrations greater than 0.4M. The logarithm of the growth rate was linearly correlated to the interfacial rigidity (inverse microviscosity) of the emulsion which suggests that coalescence rather than molecular diffusion was primarily responsible for emulsion instability under the conditions studied. PMID- 14522127 TI - Ternary naproxen: beta-cyclodextrin:polyethylene glycol complex formation. AB - The aim of this study was to investigate the effect of the presence of the water soluble polymer polyethylene glycol (PEG)-MW=35000 g/mol-on the complexation of the phototoxic anti-inflammatory drug naproxen, in its sodium salt form, with beta-cyclodextrin (beta-CD). The data revealed that the polymer does not interact with the uncomplexed naproxen whereas it does with the beta-CD. The presence of different proportions of PEG, in the 0-1% (w/w) range, systematically lowers K(app) of the formation of the naproxen:beta-CD inclusion complex. The reason for the decrease in the complexed drug is the presence of other competing equilibria, the first one is an interaction of the polymer with the beta-CD, which in turn reduces the amount of free CD available for including the naproxen, and the second is the formation of a naproxen:beta-CD:PEG ternary complex with lower affinity than the binary complex. The binding constant of these processes are K(2)=(4.5+/-1.0) x 10(5) M(-1) and K(3)=870+/-19 M(-1), respectively. In addition the presence of the PEG produces an important change in the driving force of the complex formation. In this case the process is enthalpically unfavoured and entropically favoured; these are typical characteristics of processes governed by hydrophobic interactions. PMID- 14522128 TI - Transport mechanism(s) of poly (amidoamine) dendrimers across Caco-2 cell monolayers. AB - The objective of this research was to investigate the mechanism(s) of transport of generation 2 (G2) poly (amidoamine) dendrimers across Caco-2 cell monolayers. The contribution of an energy-dependent process such as adsorptive endocytosis was investigated by determining G2 permeability at 4 and 37 degrees C. The contribution of P-gp efflux to transport was examined by determining the apical to basolateral (AB) and basolateral to apical (BA) permeability of 14C-paclitaxel in presence of G2, and by determining AB and BA permeability of G2 in presence of paclitaxel. The permeability of G2 and 14C-mannitol was investigated in the presence of palmitoyl carnitine to determine the contribution of the paracellular pathway. Permeability of G2 at 4 degrees C was significantly (P<0.05) lower than that observed at 37 degrees C. AB and BA permeability of 14C-paclitaxel did not change in the presence of G2. AB and BA permeability of G2 did not change in the presence of paclitaxel. The permeability of G2 and 14C-mannitol increased significantly (P<0.05) in the presence of palmitoyl carnitine, and in addition, 14C-mannitol permeability was increased in presence of G2. The permeability of G2 across Caco-2 cell monolayers appears to involve a combination of paracellular transport and an energy-dependent process, possibly adsorptive endocytosis. G2 dendrimers do not appear to be substrates for the P-gp efflux system. PMID- 14522129 TI - Upregulation of major histocompatibility complex class II antigens in hepatocytes in Doberman hepatitis. AB - Major histocompatibility complex (MHC) class II antigen expression in hepatocytes and its correlation with mononuclear cell infiltration into the liver were studied using immunohistochemical techniques in 38 Dobermans with Doberman hepatitis (DH). Liver biopsy samples were obtained from 18 dogs at the subclinical stage. Autopsy samples were taken from 6 DH dogs euthanized for a reason other than DH, from 14 dogs euthanized because of advanced liver failure and from 6 control Dobermans. Upon examination of the control liver samples, no expression of MHC class II antigens was detected in hepatocytes. By contrast, in 15 of the 18 DH biopsies (83%) and in all 20 DH autopsy liver samples, hepatocytes expressed MHC class II molecules. MHC class II expression was either cytoplasmic or membranous and occurred in conjunction with lymphocyte infiltration. A correlation between the inflammatory reaction and the expression of MHC class II in hepatocytes suggests that the aberrant expression of MHC class II in hepatocytes is induced by cytokines. Hepatocytes presenting a putative MHC class II molecule-associated autoantigen could thus become the target of an immune attack mediated by CD4+ T cells. In addition, corticosteroid treatment was observed to significantly decrease MHC class II expression in DH hepatocytes. Inappropriate MHC class II expression in hepatocytes and mononuclear cell infiltration are suggesting an autoimmune nature for chronic hepatitis in Dobermans. PMID- 14522130 TI - The porcine skin associated T-cell homing chemokine CCL27: molecular cloning and mRNA expression in piglets infected experimentally with Staphylococcus hyicus. AB - CCL27 (also named CTACK, ALP, ILC and ESkine) is a CC chemokine primarily expressed by keratinocytes of the skin. The cognate receptor of CCL27 named CCR10 (GPR-2), is also expressed in skin-derived cells, and in addition by a subset of peripheral blood T-cells and in a variety of other tissues. In this paper, we report the cloning of porcine CCL27 cDNA and investigation of CCL27 mRNA expression in Staphylococcus hyicus infected piglets. At the protein level, 77 and 74% homology was found to human and mouse CCL27 sequences, respectively. The results of the expression analyses show that CCL27 mRNA is upregulated in the skin of infected piglets and to a lesser extent in piglets recovered from disease and without clinical signs of infection, indicating a role for CCL27 both during inflammation and after recovery from an infection. PMID- 14522131 TI - Immunophenotype and functional properties of feline dendritic cells derived from blood and bone marrow. AB - Dendritic cells (DCs) are a heterogeneous population of cells of fundamental importance in initiating innate as well as specific immune responses. The identity and function of DCs in the cat are unknown, although they are likely pivotal in the response to infection. In this study, feline DCs were derived by 3 10-day culture of adherent blood mononuclear cells (PBMCs) and bone marrow mononuclear cells (BMMCs) in the presence of IL 4 and GM-CSF. BMMC consistently yielded a greater number of DCs than PBMC, and there were fewer macrophages than DC from both compartments. DCs expressed a distinct constellation of surface molecules, which included CD1a, CD1b, and CD1c, CD11b, CD14, and 2-3-fold higher levels of MHC class I and II molecules than co-cultured macrophages or fresh blood monocytes. DCs displayed typical cytoplasmic processes, limited non specific esterase activity, and acquired antigen by phagocytosis, pinocytosis, and binding to specific receptors. Cytokine-exposed cells induced proliferation of allogeneic lymphocytes. Thus, the cells derived by these culture conditions had markers and functions analogous to immature myeloid DCs. Availability of feline DCs will enable investigation of their role in infectious disease and their potential therapeutic application. PMID- 14522132 TI - Endometrial IL-1beta, IL-6 and TNF-alpha, mRNA expression in mares resistant or susceptible to post-breeding endometritis. Effects of estrous cycle, artificial insemination and immunomodulation. AB - Endometrial mRNA expression of the pro-inflammatory cytokines interleukin-1beta (IL-1beta), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-alpha) was assessed in mares resistant (RM) or susceptible (SM) to persistent post-breeding endometritis (PPBE). Eight RM and eight SM, were selected based on reproductive records and functional tests out of a herd of 2,000 light cross-type mares. Three experiments were done to study transcription patterns in (i) basal conditions; (ii) after artificial insemination (AI); and (iii) after administration of an immunomodulator at time of artificial insemination. Endometrial biopsies were taken during consecutive cycles: (i) at estrus, when follicles reached 35 mm and at diestrus (7 +/- 1 days after ovulation); (ii) at 24 h post-AI, with dead semen (estrus) and in diestrus; (iii) at 24 h after treatment with a Mycobacterium phlei cell-wall extract (MCWE) preparation and AI (with dead semen), and at diestrus. mRNA expression was quantitated by real time PCR. Under basal conditions, SM had significantly higher mRNA expression of all cytokines in estrus and of IL-1beta and TNF-alpha in diestrus, compared to RM. After AI, there were no differences between RM and SM in estrus; however, mRNA expression for all three pro-inflammatory cytokines was higher than under basal conditions. In diestrus, RM showed significantly lower IL-1beta and TNF-alpha mRNA expression than SM. When MCWE was administered at time of AI, no differences between cytokine induction from RM and SM were found. Globally, mRNA expression for all three cytokines correlated well among themselves when expression was high. The present study showed that (i) in basal conditions RM had lower mRNA expression of pro-inflammatory cytokines than SM with no effect of estrous cycle; (ii) AI upregulated mRNA expression for all three cytokines in both RM and SM, with persistance in diestrus in the latter; (iii) treatment with MCWE at time of AI down-regulated mRNA expression of IL-1 with significant effects in SM which behaved like RM. Immunomodulation with MCWE could be of help in restoring homeostatic local inflammatory mechanisms, thus assisting in the prophylaxis of post-breeding endometritis in mares. PMID- 14522133 TI - Antibody responses in the serum and gut of chicken lines differing in cecal carriage of Salmonella enteritidis. AB - Salmonella frequently causes human foodborne infections. Contaminated products from poultry infected with Salmonella enteritidis are mainly involved. This serovar is able to colonize the gastrointestinal tract and generally produces a chronic asymptomatic carrier state in poultry, except in very young birds. We have developed a model of S. enteritidis carriage in chicks and found that four chicken lines, B13, L2, PA12 and Y11 differ in their cecal colonization by S. enteritidis, whereas their systemic organs are similarly infected. We have monitored the serum and gut antibody responses of these four lines to S. enteritidis for 9 weeks post inoculation (pi). We confirm that S. enteritidis infected the spleens of the four chicken lines similarly, and that it often colonized the ceca at levels significantly higher in B13 and L2 chicks than those of the PA12 and Y11 chicks. The serum IgM and IgG antibody responses were high and the serum IgA antibody responses low. In contrast, the intestinal secretions contained mostly IgA antibodies. The serum IgM antibody values of the four chicken lines were similar. However, the B13 and L2 chicks often had significantly higher serum IgG and IgA antibody responses than PA12 and Y11 chicks. Only the B13 and L2 chicks showed high, persistent levels of IgA antibody in intestinal secretions. These results suggest that most antibody responses are related to cecal colonization by S. enteritidis. They also indicate that factors other than the antibody levels are involved in the control of this colonization. PMID- 14522134 TI - Protein kinase C (PKC) isotype profile in eosinophils from ponies with sweet itch and role in histamine-induced eosinophil activation. AB - Eosinophils have been implicated in the pathogenesis of the seasonal equine allergic skin disease, sweet itch. Protein kinase C (PKC) is involved in regulating eosinophil function and antigen challenge has been reported to alter PKC isotype expression in blood eosinophils from allergic human subjects. Here we have compared the pattern of PKC isotype expression in eosinophils from sweet itch ponies with that in cells from normal ponies both during the active and inactive phases of the disease. A role for PKC in histamine-induced eosinophil activation was also investigated. Conventional PKCs alpha and beta, novel PKCs delta and epsilon and atypical PKCs iota and zeta were identified in eosinophils pooled from four allergic ponies during the inactive phase, when no clinical signs were evident. The PKC isotypes, like those in eosinophils from normal ponies, were located primarily in the particulate fraction of the cell. Isotype expression in cells from normal and allergic animals did not appear to be different. In contrast, during the active phase of the disease, when the sweet itch ponies had clinical signs, the expression of PKCs beta, epsilon and iota in eosinophils from these animals appeared to be increased relative to that in cells from normal ponies. When PKC expression in eosinophils from five individual normal and sweet itch ponies was compared, small, but statistically significant, increases in PKC epsilon and PKCdelta expression were evident in eosinophils from the sweet itch ponies during the active and inactive phases, respectively. The non-selective PKC inhibitors, staurosporine and Ro31-8220, significantly reduced histamine-induced superoxide production. Use of Go6976, an inhibitor of conventional PKCs, suggested that PKCalpha and/or beta were involved and that there was significantly greater inhibition of the response in eosinophils obtained from sweet itch ponies during the active phase. There was no significant difference in histamine-induced superoxide production by eosinophils from allergic and normal ponies and the functional significance of the increased PKC isotype expression in eosinophils from sweet itch ponies relative to that in cells from healthy animals remains to be established. PMID- 14522135 TI - Cross-reactivity of human and bovine antibodies in striped dolphin paraffin wax embedded tissues. AB - This study evaluates the cross-reactivity of seven anti-human and one anti-bovine antibodies in formalin-fixed, paraffin-embedded tissue samples of liver and mesenteric lymph nodes of 13 striped dolphins (Stenella coeruleoalba). Four antibodies (CD3, IgG, lysozyme and S100 protein) reacted with striped dolphin lymph nodes in a similar pattern to that observed in the species of origin. The anti-human MHC class II mAb reacted strongly with macrophages and dendritic-like cells of striped dolphins, whereas a small number of lymphocytes were labelled with this antibody. These antibodies were used to study the immunophenotype of the inflammatory infiltrated in non-specific chronic reactive hepatitis (eight cases) and chronic parasite cholangitis (two cases) and normal liver (three cases) of striped dolphins. Non-specific chronic reactive hepatitis was composed of inflammatory infiltration of CD3+ T lymphocytes and IgG+ plasma cells in portal spaces and hepatic sinusoids. Lymphonodular aggregates observed in chronic parasitic cholangitis showed a cellular distribution similar to that found in lymph node cortex, including the presence of S100+ and MHC class II+ dendritic like cells in lymphoid follicles and interfollicular areas. This result suggests that those inflammatory infiltrates are highly organised to enhance antigen presentation to B and T cells. PMID- 14522136 TI - Development of an enzyme immuno assay for the determination of porcine haptoglobin in various body fluids: testing the significance of meat juice measurements for quality monitoring programs. AB - Quantification of haptoglobin (Hp), an acute phase protein, in blood is presently discussed as being useful to monitor animal health. We developed an enzyme immuno assay (EIA) which is specific for porcine Hp, is not impaired by hemolytic samples and is sufficiently sensitive to be applied in meat juice. Hp was purified from porcine serum by affinity chromatography on hemoglobin Sepharose followed by gel filtration. A specific rabbit antiserum was obtained. In a competitive approach, biotinylated porcine Hp was used as tracer and incubated with Hp standard or sample in microtiter plates. The limit of detection was 0.02 mg/l, parallelism of sample dilutions was proven; recovery of Hp added to serum samples was 96.4 +/- 4.7%. The coefficients of intra and inter-assay variation were 3.3 (n=5) and 10.2% (n=16), respectively. Hp was reliably quantified in blood serum and plasma, whole blood, saliva and meat juice. For healthy pigs of different ages (4 weeks and 6 months), mean Hp concentrations of about 0.5-0.7 mg/ml were observed. To test the significance of Hp measurements in other matrices, samples were obtained from fattening pigs or from slaughter pigs. Blood serum or plasma was collected in parallel. In whole blood, Hp concentrations were about 40% lower than in plasma, but were closely related (n=24,r=0.85,P<0.001). Saliva Hp concentrations ranged between 0.3 and 3.0 microg/ml and were marginally related with blood plasma concentrations (n=93,r=0.35,P<0.001). From 106 hybrid slaughter pigs (100-110 kg) blood and muscle samples (diaphragmatic pillar, d.p.; m. brachiocephalicus, m.b.) were collected. Meat juice was obtained after freezing and thawing. Concentrations were 0.39+/-0.5 mg/ml in serum and 0.04+/ 0.06 mg/ml in meat juice. Hp concentrations in blood were closely correlated with those in d.p. juice (P<0.001,r=0.750) and m.b. juice (P<0.001,r=0.776). In view of the many reports on Hp measurements being predictive for animal health even in the subclinical range, we conclude that Hp quantification in meat juice might be useful to assess meat quality at slaughter and further along the processing chain in terms of animal health. PMID- 14522137 TI - Recurrent airway obstruction (RAO) in horses is characterized by IFN-gamma and IL 8 production in bronchoalveolar lavage cells. AB - In horses prone to developing recurrent airway obstruction (RAO), we tested the hypotheses that the cytokine profile in the bronchoalveolar lavage (BAL) cells of affected horses would reflect a polarized Th-2 response; that cytokine and chemokine alterations would occur within 24 h of allergen exposure; and that allergen exposure would induce alterations in the expression of the transcription factor t-bet (t-box-expressed in T-cells). The expression levels of interleukin-4 (IL-4), IL-13, Interferon-gamma (IFN-gamma), t-bet, IL-8 and granulocyte macrophage colony stimulating factor (GM-CSF) were measured in BAL cells obtained from control and RAO-susceptible horses during an asymptomatic phase and at 24 h and 5 weeks post-stabling and hay exposure. At each sampling time, BAL neutrophil percentages in the RAO-group exceeded controls. In the RAO-group, only IL-13 expression was decreased 2-fold during the asymptomatic phase. No differences in cytokine or chemokine expression were detected during the acute exposure phase. During the chronic phase, IFN-gamma and IL-8 expression levels were 2.5- and 3 fold greater, respectively, in the RAO-group. No other differences in gene expression were detected. We conclude that the cytokine profile of the airway cells does not reflect a polarized Th-2 response; that increases in IFN-gamma result from a t-bet independent pathway and that chemokines from epithelial or interstitial cells may contribute to early neutrophil influx. PMID- 14522138 TI - Characterization of a spontaneously transformed chicken mononuclear cell line. AB - We describe the characterization of a spontaneously transformed chicken monocytic cell line that developed as a single colony of cells in a heterophil culture that was inadvertently left in the incubator over a period of 25 days. These cells, hitherto named HTC, grow efficiently at both 37 or 41 degrees C in culture medium containing either 5% FBS or 2% chicken serum. The HTC cells are acid phosphatase positive, show expressions of both class I and class II major histocompatibility complex (MHC), CD44, K1, and K55 cell surface antigens, and engulf latex beads, produce nitrite and interleukin-6 on stimulation with bacterial lipopolysaccharide (LPS). Treatment with phorbol myristate acetate (PMA) induces respiratory burst in HTC cells and the secretion of matrix metalloproteinase (MMP) into culture medium. Using gene-specific primers and reverse transcriptase polymerase chain reaction (RT-PCR), the presence of mRNA trancripts for interferon-gamma (IFN-gamma), interleukin-1 (IL-1), interleukin-6 (IL-6), nitric oxide synthase (NOS), matrix metalloproteinase-2 (MMP-2), and transforming growth factor-beta (TGF-beta) were detected. Lipopolysaccharide (LPS) treatment of HTC cells modulated IL-1, IL-6, IFN-gamma, NOS mRNA levels as detected by RT-PCR analyses. Using different avian tumor virus gene-specific primers and PCR, the HTC cells were positive for the presence of avian leukosis virus (ALV) and Marek's disease virus (MDV) but negative for reticuloendothelial virus (REV), chicken infectious anemia virus (CIAV), and herpes virus of turkeys (HVT). The production of ALV antigens by HTC cells was further confirmed using p27 gag protein ELISA. Collectively, these results show that the HTC cells belong to myeloid/macrophage lineage and were likely transformed by ALV and MDV but retain many interesting and useful biological activities. PMID- 14522139 TI - High level expression, purification, and in vivo activity of bovine granulocyte colony stimulating factor produced using a baculovirus system. AB - A bovine granulocyte-colony stimulating factor (bG-CSF) cDNA clone bearing a C terminal poly-His-tag (bG-CSFHis) was constructed and expressed by the baculovirus expression system. The bG-CSFHis was expressed as an approximately 19kDa protein in the culture supernatants and was purified using a nickel chelate column. The purified bG-CSFHis had bioactivity in vitro in the NFS-60 bioassay. In order to evaluate activity in vivo, purified bG-CSFHis was administered to cattle as single or multiple dosages. The bG-CSFHis increased neutrophil counts in peripheral blood and modulated the phagocytic activity of the neutrophils. The data indicates that the recombinant protein had activity in vivo. PMID- 14522140 TI - Quantitative changes in the normal and apoptotic thymocytes of pigs treated with anabolic doses of the beta2 adrenergic agonist clenbuterol. AB - Although the beta2 adrenergic agonists have been seen to have important effects on the mechanisms regulating the development and death of T-cells in the thymus, the side-effects on the immune system of anabolic treatments of these substances have hardly been considered. In order to evaluate the effects exerted by the beta2 adrenergic agonist clenbuterol on the thymocyte population, the thymus of eight pigs treated with anabolic doses of this substance was studied by morphometric methods, regarding apoptotic (terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL)-positive) and normal (TUNEL-negative) cells. The thymus of another eight pigs fed without clenbuterol served as a control. The clenbuterol treatment had a clear effect on the thymocyte size, decreasing their mean nuclear area. The T-cell apoptosis index was also affected by the clenbuterol, significantly increasing the apoptosis percentage in the treated group with respect to the control. In the light of our results, the clenbuterol induced thymocyte apoptosis throughout the thymus and caused morphometric changes in the thymocyte population, which was in line with the immunosuppressive role attributed to other beta2 adrenergic agonists. PMID- 14522141 TI - Neurons that respond to more than one depth cue. AB - The 3D orientation of a surface can be specified by perspective, motion parallax or binocular disparity. Tsutsui et al. have found cells in the monkey intraparietal sulcus that responded to surface orientation defined only by a texture gradient. Most of these cells also responded to orientation defined only by binocular disparity. PMID- 14522142 TI - Alpha-synuclein oligomerization: a role for lipids? AB - Alpha-Synuclein is a core component of the proteinaceous aggregates observed in several neurodegenerative diseases. A central role of alpha-synuclein in neurodegeneration was demonstrated by the discovery of missense alpha-synuclein mutations in familial Parkinson's disease. However, the specific mechanism by which alpha-synuclein contributes to these diseases remains unclear. A recent study by Sharon et al. linked the presence of specific fatty acids to the appearance of alpha-synuclein oligomers in vivo. alpha-Synuclein oligomers might be a first step in the formation of alpha-synuclein aggregates present in a number of neurodegenerative diseases, although their cytotoxicity remains to be directly demonstrated. PMID- 14522143 TI - New roles for astrocytes (stars at last). AB - Astrocytes have been presumed to be uninteresting for the better part of a century. Recent findings suggest unexpected new functions for these cells and highlight the importance of viewing most brain activities as a collaboration between astrocytes and neurons. Astrocytes have been implicated in dynamic regulation of neuron production, synaptic network formation, neuron electrical activity and specific neurological diseases. We are only at the threshold of understanding fully the nature and consequences of these new astrocyte functions, and there is still much to be learned about the older and better-known roles. But now, at last, astrocytes have our attention. PMID- 14522144 TI - New roles for astrocytes: redefining the functional architecture of the brain. AB - Astrocytes have traditionally been considered ancillary, satellite cells of the nervous system. However, work over the past decade has revealed that they interact with the vasculature to form a gliovascular network that might organize not only the structural architecture of the brain but also its communication pathways, activation, thresholds and plasticity. The net effect is that astroglia demarcate gray matter regions, both cortical and subcortical, into functional compartments whose internal activation thresholds and external outputs are regulated by single glial cells. The array of these astrocyte-delimited microdomains along the capillary microvasculature allows the formation of higher order gliovascular units, which serve to match local neural activity and blood flow while regulating neuronal firing thresholds through coordinative glial signaling. By these means, astrocytes might establish the functional as well as the structural architecture of the adult brain. PMID- 14522145 TI - New roles for astrocytes: regulation of CNS synaptogenesis. AB - The notion that astrocytes have a profound influence on the function of synapses between CNS neurons implies that the development of synaptic connections and their glial neighbors are controlled by reciprocally acting signals. Currently, however, synaptogenesis is considered a purely neuronal affair. This article summarizes recent experimental evidence suggesting that this may not be the case. Astrocytes may indeed regulate the formation, maturation and maintenance of synapses. The recent advances caution that synapses cannot develop correctly without astrocytes. Further progress on this issue requires new experimental models to identify signaling pathways and to scrutinize the relevance of glia synapse interactions in vivo. PMID- 14522146 TI - New roles for astrocytes: regulation of synaptic transmission. AB - Abstract Although glia often envelop synapses, they have traditionally been viewed as passive participants in synaptic function. Recent evidence has demonstrated, however, that there is a dynamic two-way communication between glia and neurons at the synapse. Neurotransmitters released from presynaptic neurons evoke Ca2+ concentration increases in adjacent glia. Activated glia, in turn, release transmitters, including glutamate and ATP. These gliotransmitters feed back onto the presynaptic terminal either to enhance or to depress further release of neurotransmitter. Transmitters released from glia can also directly stimulate postsynaptic neurons, producing either excitatory or inhibitory responses. Based on these new findings, glia should be considered an active partner at the synapse, dynamically regulating synaptic transmission. PMID- 14522147 TI - Malignant gliomas: perverting glutamate and ion homeostasis for selective advantage. AB - The malignant transformation of astrocytes, oligodendrocytes or their progenitor cells gives rise to tumors that are collectively called gliomas. Although the vast majority of these tumors are believed to be of astrocytic origin, the actual cell of origin remains unknown. And whereas gliomas present with some genetic alterations in tumor-suppressor genes or oncogenes that are common among cancers, their biology differs significantly from other neoplasms. Most notably, they grow in a confined space defined by the bones of the skull and spinal column, and they spread by active cell migration rather than by passive, humorous spread. Recent research suggests that astrocyte-derived tumors co-opt neurotransmitters, their transporters and ion channels to support their unusual growth and invasive migration. PMID- 14522149 TI - Inflammation, degeneration and regeneration in the injured spinal cord: insights from DNA microarrays. AB - GeneChip microarrays have recently been introduced to the field of neurobiology to identify and monitor the expression levels of thousands of genes simultaneously. This powerful technique is now used for studying the pathophysiology of CNS injuries including spinal cord lesions. Early stages after injury are characterized by the strong upregulation of genes involved in transcription and inflammation and a general downregulation of structural proteins and proteins involved in neurotransmission. Later, an increase in the expression of growth factors, axonal guidance factors, extracellular matrix molecules and angiogenic factors reflects the attempts for repair, while upregulation of stress genes and proteases and downregulation of cytoskeletal and synaptic mRNA reflect the struggle of the tissue to survive. DNA microarrays have the potential to aid discovery of new targets for neuroprotective or restorative therapeutic approaches PMID- 14522150 TI - Interneuron Diversity series: Hippocampal interneuron classifications--making things as simple as possible, not simpler. AB - The nervous system is made up of many specific types of neuron intricately intertwined to form complex networks. Identifying and defining the characteristic features of the many different neuronal types is essential for achieving a cellular understanding of complex activity from perception to cognition. So far, cortical GABAergic interneurons have represented the epitome of cellular diversity in the CNS. Despite the desperate need for effective classification criteria allowing a common language among neuroscientists, interneurons still evoke memories of Babel. Several approaches are now available to overcome the challenges and problems associated with the various classification systems used so far. PMID- 14522148 TI - The multiple personalities of h-channels. AB - Concepts regarding the function of the hyperpolarization-activated current (Ih) in shaping the excitability of single cells and neuronal ensembles have been evolving rapidly following the recent cloning of genes that encode the underlying 'h-channels' - the HCN genes. This article reviews new information about the transcriptional regulation of these channels, highlighting novel studies that demonstrate short- and long-term modulation of HCN expression, and linking this modulation to mechanisms of neurological diseases. PMID- 14522151 TI - Inequalities or disparities in health--initiatives separated by a common language? PMID- 14522152 TI - The effect of work environment and heavy smoking on the social inequalities in smoking cessation. AB - OBJECTIVES: To investigate social differences with regard to smoking cessation in Denmark. METHODS: Social differences in smoking cessation were estimated from 3606 smokers from the Danish National Work Environment Cohort Study in 1990, 1995 and 2000. The differences were investigated using heavy smoking and four work environment factors as explanatory variables. RESULTS: The odds ratio for cessation was more than twice as high in social class I compared with social class V. When heavy smoking was controlled, this explained 28% of social differences, the work-environment factors alone explained 36% of social differences, and together the factors explained 55% of social differences. CONCLUSIONS: A large proportion of the social differences in smoking cessation could be explained by differences in work-environment exposures and smoking intensity. PMID- 14522153 TI - Colorectal cancer, screening and survival: the influence of socio-economic deprivation. AB - OBJECTIVES: To determine the extent to which socio-economic deprivation explains colorectal cancer prevalence, subject participation in screening, and postoperative survival and life expectancy. METHODS: Regression analyses of clinical data from a large randomized controlled trial, augmented by geographical based indices of deprivation. RESULTS: Deprivation appears to exert no significant impact on colorectal cancer prevalence but is a major factor explaining subject participation in screening. Cancer detection at later stages reduces life expectancy at time of treatment. Females from more-deprived areas have poorer post-treatment life expectancies and survival prospects, independently of their screening behaviour. CONCLUSIONS: Screening increases the chances of having a cancer treated at an earlier stage, and treatment at an earlier stage is associated with longer subsequent life expectancy. However, those from more-deprived areas are less likely to accept an invitation to be screened. PMID- 14522154 TI - Possibilities and barriers in the implementation of a preconceptional screening programme for cystic fibrosis carriers: a focus group study. AB - OBJECTIVE: This qualitative study aimed to explore possibilities and barriers in the implementation of a nationwide preconceptional cystic fibrosis (CF) carrier screening programme. METHODS: Sessions were held with two focus groups of CF patients and CF relatives, one focus group of people from the target population (couples planning a pregnancy), and two focus groups of potential providers (general practitioners (GPs) and municipal health service workers). RESULTS: Important barriers in the implementation of a preconceptional CF carrier screening programme included the problem of reaching the target population, the heavy workload of GPs, the limited public knowledge about CF in general, and the absence of a preconceptional consultation setting. In general, there was a positive attitude among the participants towards CF carrier screening. CONCLUSION: This study revealed some important barriers in the implementation of CF carrier screening programmes. More research is needed to specify and quantify the importance of the various barriers. Eventually, different intervention strategies should be included in an implementation plan to overcome the most important barriers in the organization and execution of screening. PMID- 14522155 TI - Unintentional home injury in preschool-aged children: looking for the key--an exploration of the inter-relationship and relative importance of potential risk factors. AB - OBJECTIVE: To investigate the physical, social and psychological environment of families with preschool-age children to identify the most significant risk factors for unintentional injury. DESIGN: A 1-year prospective case-control study, using a health-visitor-administered questionnaire. SETTING: East and Midlothian, Scotland. SUBJECTS: Seventy-nine children under 5 years of age presenting to an accident and emergency (A&E) department during 1998-1999 with an unintentional home injury and 128 matched controls. RESULTS: Of 264 families, 207 responded (78.4% response rate). The main carers of cases had a lower level of educational attainment than controls (P<0.01). This factor explained the case carer leaving fulltime education earlier, being less likely to be married and more often in receipt of government benefits. Cases lived in households with larger numbers of children, were more likely to have a physical illness, were less likely to have had a non-medically attended injury in the previous year (P<0.01) but more likely to have had another A&E injury attendance. Case households had lower electrical socket cover utilization (P<0.01) and fewer thought their child had adequate access to safe play areas. The main carers of cases tended to have a more negative life event experience in the preceding 6 months, but showed no significant differences in physical or mental well-being or social support. Cases seemed to be slightly more deprived members of their community. CONCLUSIONS: The main carer's educational attainment and socket cover utilization were lower in case families. These risk factors could be used to target families for injury-prevention work. Initiatives to raise educational achievement in the general population could lead to reductions in childhood injuries. PMID- 14522156 TI - The health needs of children in public care: the results of an audit of immunizations of children in care. AB - An immunization audit was conducted to assess the health status of children in public care in the boroughs of Middlesbrough and Redcar & Cleveland. The audit found that children in public care have lower immunization rates and more incomplete schedules than the local population. Obtaining health data about these children can be problematic. For purposes of accuracy, it is important to refer to all the child's health records. Some children who were initially thought to have incomplete schedules had completed their immunizations, whereas others who were thought to be up to date had incomplete schedules. PMID- 14522157 TI - The use of primary/national school absenteeism as a proxy retrospective child health status measure in an environmental pollution investigation. AB - As a result of community concerns over animal and human ill health centred around a rural town in the Republic of Ireland, a series of epidemiological studies were conducted. The absence of adequate health-information systems forced researchers to investigate alternative methods of assessing child health in the 'at-risk' area. This study aimed to examine annual primary/national school attendance data over a 10 year period as a proxy health status measure. Data from six geographical areas were analysed; one of these six areas was designated the 'high risk' area on the basis of reports of animal ill health. Significantly higher absenteeism rates were noted in the 'high-risk' area in nine of the 10 years examined. Although caution is urged in the interpretation of these results, this study demonstrates that primary/national school absenteeism data can act as a useful, albeit crude, proxy measure of health status. PMID- 14522158 TI - Regional differences in healthy life expectancy in The Netherlands. AB - BACKGROUND: Healthy life expectancy has mainly been studied at the level of healthcare systems rather than at regional level within healthcare systems. In this article, healthy life expectancy at birth and at 65 years of age for men and women in the Netherlands has been described, and factors related to these regional variations have been explored. METHODS: Ecological study of 27 healthcare regions (hospital catchment areas). Life expectancy and healthy life expectancy were calculated using 1995 mortality data and pooled health interview survey data (1992-1997) from Statistics Netherlands. RESULTS: Healthy life expectancy shows a regional pattern, slightly different from that found in life expectancy and self-reported health. The regional distribution of male and female healthy life expectancy is different, especially at 65 years. Healthy life expectancy of women aged 65 years is independent of their total life expectancy. Social conditions and lifestyle differences between regions are negatively associated with healthy life expectancy in Dutch regions. Healthcare supply variables show no clear relationship. CONCLUSION: Although the Netherlands is a small, homogeneous country, substantial differences were found in healthy life expectancy. PMID- 14522159 TI - Prevalence and determinants of obesity in an urban sample of Portuguese adults. AB - OBJECTIVE: To evaluate the prevalence and the determinants of obesity, and the associated cardiovascular risk factors in a random sample of non institutionalised adults. DESIGN: Cross-sectional study. SUBJECTS: A random sample of 1436 habitants of Porto (873 women and 563 men) aged 18-90 years. MEASUREMENTS: All participants answered a structured questionnaire comprising information on social, demographic, behavioural and clinical aspects. Anthropometric measures, blood pressure and fasting blood samples were obtained. Diet was assessed using a semi-quantitative food-frequency questionnaire, and physical activity was evaluated using a questionnaire exploring all professional, domestic and leisure-time activities. When the body mass index was > or =30 kg/m2, the subject was considered as 'obese'. Proportions were age adjusted for the European population. Odds ratios and 95% confidence intervals were computed using unconditional logistic regression. RESULTS: The prevalence of obesity was significantly higher in women (26.1%) than men (13.9%). Regardless of gender, obesity increased with age, decreased with education, and was more frequent in married blue-collar workers and unemployed subjects. Smoking was more prevalent in subjects of normal weight, and a higher proportion of those reporting no regular exercise were obese. In men, obesity prevalence increased with increasing quartiles of energy intake, but no such changes were found in women. Compared with subjects of normal weight, obese men showed a significantly higher prevalence of hypertension (53.3 vs 26.1%) and hypertriglyceridaemia (23.4 vs 9.0%). Also, hypertension (43.7 vs 30.7%), diabetes (7.6 vs 2.7%), hypertriglyceridaemia (27.1 vs 5.0%), and abnormal low-density lipoprotein (30.4 vs 21.4%) and high-density lipoprotein cholesterol concentration (15.0 vs 5.3%) were more frequent in obese women. CONCLUSION: Obesity is a major public health issue in urban Portuguese populations, and obese individuals have many features of metabolic syndrome. Education and relative deprivation are modifiable factors that are significantly associated with obesity. However, no clear-cut relationship was found between physical activity and energy intake. PMID- 14522160 TI - Can patients with osteoporosis, who should benefit from implementation of the national service framework for older people, be identified from general practice computer records? A pilot study that illustrates the variability of computerized medical records and problems with searching them. AB - Although UK general practice is highly computerized, comprehensive use of these computers is often limited to registration data and the issue of repeat prescriptions. The recording of diagnostic data is patchy. This study examines whether patients with, or at risk of, osteoporosis can be readily identified from general practice computer records. It reports the findings of a pilot study designed to show the variability of recording the diagnosis of osteoporosis and osteopenia, as well as how useful surrogate markers might be to identify these patients. The study also illustrates the difficulties that even skilled practitioners in a primary care research network experience in extracting clinical data from practice information systems. Computer searches were carried out across six practices in a general practice research network in the south-east of England. Two of these practices had previously undertaken research projects in osteoporosis and were consequently expected to have excellent data quality in osteoporosis. These two practices had a combined list size of 27,500 and the remaining practices had a combined practice population of 43,000 patients. The data were found to be variable with over 10-fold differences between practices in the recorded prevalence of osteoporosis diagnosis as well as its surrogate markers-such as fragility fractures, long-term steroid prescription, etc. There was no difference in data quality between the two practices that had conducted osteoporosis research and the rest of the group, other than in the areas of diagnostic recording and prescribing for osteoporosis and recording of fractures. Issues were raised by the practices that struggled to identify patients at risk of osteoporosis about the limitations of Read classification in this disease area. Practices need further assistance if the patients at risk are to be identified. Without urgent action, it will be difficult for practices to identify the patients who are likely to benefit from Standard 6-'Falls' of the National Service Framework for Older People. These findings also have broader implications as UK general practice moves towards the implementation of a quality-based contract. PMID- 14522161 TI - Pox in the docks: varicella outbreak in an Australian prison system. AB - OBJECTIVES: 1. Describe an outbreak of varicella in a prison system. 2. Highlight the risks of disease transmission within the prison environment. 3. Promote infection control guidelines for high-risk sub-groups within the prison system, including the application of quarantine. SETTING: Four prisons, one prison hospital, the prison transport system, one courthouse. MAIN OUTCOME MEASURES: Number of cases of varicella infection; reported varicella immunity status of cases and contacts; immunity status of known HIV antibody positive inmates. RESULTS: Five cases of chickenpox were identified. There were 23 contacts of the Index Case occurring during transport between prison and court and whilst being held in the court holding cells. Two of these contacts developed chickenpox despite having given a prior history of infection. There were over 300 inmates exposed to varicella zoster virus (VZV) during the outbreak, including one HIV antibody positive inmate who had serologically confirmed immunity. This inmate developed shingles following exposure to VZV from one of the cases. CONCLUSIONS: There is an elevated risk of respiratory transmission of infections such as chickenpox in prisons. Clear guidelines should be in place to protect HIV antibody positive people, pregnant women, and others who are at increased risk of complications from such infections. In the case of varicella, all inmates and staff without documented immunity should be screened to determine immunity, and if non-immune, should be offered VZV vaccination. Every effort should be made to prevent HIV antibody positive inmates being exposed to varicella, regardless of their varicella immunity status. If an HIV antibody positive inmate, who is known to be non-immune is exposed to varicella, Varicella Zoster immunoglobulin should be given within 96 h. PMID- 14522162 TI - Water, sanitation and hygiene for the health of developing nations. AB - The water and sanitation needs of the poor in developing countries are huge. To meet the target of water and sanitation for all by the end of 2025, some 2.9 billion people will have to receive improved water supplies, and 4.2 billion improved sanitation. The technologies used must be appropriate and, in particular, simple, affordable and sustainable. PMID- 14522167 TI - Combining endogenous and exogenous spatial variability in analytical population models. AB - Analytically tractable models of dynamics in continuous space rarely incorporate both endogenous and exogenous spatial heterogeneity. We use spatial moment equations in combination with simulation models to analyze the combined effects of endogenous and exogenous variability on population viability in a simple single-population model where landscape heterogeneity and local population density both affect mortality rate. The equations partition the effects of heterogeneity into an effect of local crowding and an effect of habitat association caused by differential mortality. Exogenous heterogeneity in mortality rate increases population viability through habitat association and decreases it through increased crowding; the net effect of exogenous heterogeneity is generally to improve population viability. This result is contrary to some (but not all) conclusions in the literature, which usually focus on the effects of fragmentation rather than the benefits of refuges to short dispersing individuals. PMID- 14522168 TI - Pair-edge approximation for heterogeneous lattice population models. AB - To increase the analytical tractability of lattice stochastic spatial population models, several approximations have been developed. The pair-edge approximation is a moment-closure method that is effective in predicting persistence criteria and invasion speeds on a homogeneous lattice. Here we evaluate the effectiveness of the pair-edge approximation on a spatially heterogeneous lattice in which some sites are unoccupiable, or "dead". This model has several possible interpretations, including a spatial SIS epidemic model, in which some sites are occupied by immobile host-species individuals while others are empty. We find that, as in the homogeneous model, the pair-edge approximation is significantly more accurate than the ordinary pair approximation in determining conditions for persistence. However, habitat heterogeneity decreases invasion speed more than is predicted by the pair-edge approximation, and the discrepancy increases with greater clustering of "dead" sites. The accuracy of the approximation validates the underlying heuristic picture of population spread and therefore provides qualitative insight into the dynamics of lattice models. Conversely, the situations where the approximation is less accurate reveals limitations of pair approximation in the presence of spatial heterogeneity. PMID- 14522169 TI - Metapopulation-level adaptation of insect host plant preference and extinction colonization dynamics in heterogeneous landscapes. AB - Species living in highly fragmented landscapes typically occur as metapopulations with frequent turnover of local populations. The turnover rate depends on population sizes and connectivities, but it may also depend on the phenotypic and genotypic composition of populations. The Glanville fritillary butterfly (Melitaea cinxia) in Finland uses two host plant species, which show variation in their relative abundances at two spatial scales: locally among individual habitat patches and regionally among networks of patches. Female butterflies in turn exhibit spatial variation in genetically determined host plant preference within and among patch networks. Emigration, immigration and establishment of new populations have all been shown to be strongly influenced by the match between the host plant composition of otherwise suitable habitat patches and the host plant preference of migrating butterflies. The evolutionary consequences of such biased migration and colonization with respect to butterfly phenotypes might differ depending on spatial configuration and plant species composition of the patches in heterogeneous patch networks. Using a spatially realistic individual based model we show that the model-predicted evolution of host plant preference due to biased migration explains a significant amount of the observed variation in host plant use among metapopulations living in dissimilar networks. This example illustrates how the ecological extinction-colonization dynamics may be linked with the evolutionary dynamics of life history traits in metapopulations. PMID- 14522170 TI - Modeling biological invasions into periodically fragmented environments. AB - Range expansion of a single species in a regularly striped environment is studied by using an extended Fisher model, in which the rates of diffusion and reproduction periodically fluctuate between favorable and unfavorable habitats. The model is analyzed for two initial conditions: the initial population density is concentrated on a straight line or at the origin. For each case, we derive a mathematical formula which characterizes the spatio-temporal pattern of range expansion. When initial distribution starts from a straight line, it evolves to a traveling periodic wave (TPW), whose frontal speed is analytically determinable. When the range starts from the origin, it tends to expand radially at a constant average speed in each direction (ray speed) keeping its frontal envelope in a similar shape. By examining the relation between the ray speed and the TPW speed, we derive the ray speed in a parametric form, from which the envelope of the expanding range can be predicted. Thus we analyze how the pattern and speed of the range expansion are affected by the pattern and scale of fragmentation, and the qualities of favorable and unfavorable habitats. The major results include: (1). The envelope of the expanding range show a variety of patterns, nearly circular, oval-like, spindle-like, depending on parameter values; (2). All these patterns are elongated in the direction of stripes; (3). When the scale of fragmentation is enlarged without changing the relative spatial pattern, the ray speed in any direction increases, i.e., the rate of range expansion increases. PMID- 14522171 TI - Establishment success and extinction risk in autocorrelated environments. AB - We consider establishment success (and extinction risk of small populations) in fluctuating environments, by means of an inhomogeneous branching process model. In this model it is assumed that individuals reproduce asexually during discrete reproduction periods. Within each period individuals reproduce independently and have random numbers of offspring. Expected numbers of offspring vary over reproduction periods due to random environmental changes. Previous simulation results indicated that there is a positive autocorrelation between the establishment probabilities of invaders in successive reproduction periods when environmental states are independently distributed. This result was never formally proved. In this paper we prove that this is indeed true, regardless of the form of the distribution of environmental states or the offspring distribution (under a monotonicity condition, which holds for biologically realistic models). Furthermore, we prove that it is also true for positively autocorrelated environmental states. We show by a counterexample that in environments with a strong negative autocorrelation establishment probabilities can be negatively autocorrelated. This was further examined through simulations. Our results imply that in independent, positively autocorrelated and weakly negatively autocorrelated environments the probability of success of invasion in different independently varying sites is the highest, followed by sequential invasion. For environments with a strong negative autocorrelation, sequential invasion has the highest probability of success. Effects of autocorrelation were further examined with simulations. From the results it appears that the expected length of 'runs of bad luck' is the most crucial factor for establishment success. PMID- 14522172 TI - Impacts of environmental variability in open populations and communities: "inflation" in sink environments. AB - Ecological communities are typically open to the immigration and emigration of individuals, and also variable through time. In this paper we argue that interesting and potentially important effects arise when one splices together spatial fluxes and temporal variability. The particular system we examine is a sink habitat, where a species faces deterministic extinction but is rescued by recurrent immigration. We have shown, using a simple extension of the canonical exponential growth model in a time-varying environment, that variation "inflates" the average abundance of sink populations. We can analytically quantify the magnitude of this effect in several special cases (square-wave temporal variation and Gaussian stochastic variation). The inflationary effect can be large in "intermittent" sinks (where there are periods with positive growth), and when temporal variation is strongly autocorrelated. The effect appears to be robust to incorporation of demographic stochasticity (due to discrete birth-death immigration processes), and to direct density dependence. With discrete generations, however, one can observe a wide range of effects of temporal variation, including depression as well as inflation. We argue that the inflationary effect of temporal variation in sink habitats can have important implications for community structure, because it can increase the average abundance (and hence local impacts) of species that on average are being excluded from a local community. We illustrate the latter effect using a familiar model of exploitative competition for a single limiting resource. We demonstrate that temporal variation can reverse local competitive dominance, even to the extent of allowing an inferior competitor maintained by immigration to exclude a competing species that would be locally superior in a constant environment. PMID- 14522173 TI - The age-structured lottery model. AB - The lottery model of competition between species in a variable environmental has been influential in understanding how coexistence may result from interactions between fluctuating environmental and competitive factors. Of most importance, it has led to the concept of the storage effect as a mechanism of species coexistence. Interactions between environment and competition in the lottery model stem from the life-history assumption that environmental variation and competition affect recruitment to the adult population, but not adult survival. The strong role of life-history attributes in this coexistence mechanism implies that its robustness should be checked for a variety of life-history scenarios. Here, age structure is added to the adult population, and the results are compared with the original lottery model. This investigation uses recently developed shape characteristics for mortality and fecundity schedules to quantify the effects of age structure on the long-term low-density growth rate of a species in competition with its competitor when applying the standard invasibility coexistence criterion. Coexistence conditions are found to be affected to a small degree by the presence of age structure in the adult population: Type III mortality broadens coexistence conditions, and type I mortality makes them narrower. The rates of recovery from low density for coexisting species, and the rates of competitive exclusion in other cases, are modified to a greater degree by age structure. The absolute rates of recovery or decline of a species from low density are increased by type I mortality or early peak reproduction, but reduced by type III mortality or late peak reproduction. Analytical approximations show how the most important effects can be considered as simple modifications of the long-term low-density growth rates for the original lottery model. PMID- 14522174 TI - Quantifying and testing coexistence mechanisms arising from recruitment fluctuations. AB - Temporal fluctuations in recruitment are involved in two distinct coexistence mechanisms, the storage effect and relative nonlinearity of competition, which may act simultaneously to stabilize species coexistence. It is shown that comparisons of recruitment variation between species at high versus low densities can test whether these mechanisms are responsible for stable coexistence. Moreover, under certain circumstances, these comparisons can measure the total coexistence stabilizing effect of the mechanism. These comparisons are clearest for the situation of an invader (a species perturbed to low density) in the presence of its competitors, termed residents. Then average invader-resident differences in the variances of log recruitment, potentially weighted by adult survival rates and species' sensitivities to competition, are proportional to the overall stabilizing effect of the storage effect and relative nonlinearity of competition. Less effective comparisons are available for species naturally at high and low densities or with substantial mean differences in average fitness. These developments lead also to a technique of partitioning the long-term low density growth rate of a species into community average measures of stabilizing mechanisms, deviations from these measures, and other factors. The community average measure is argued as most appropriate for understanding the ability of a coexistence mechanism to stabilize coexistence. Individual species' deviations from the community average indicate the ways in a which a coexistence mechanism may affect average fitness differences between species either enhancing or diminishing the ability of a given set of species to coexist, depending on other factors. This approach provides a general new tool for analyzing species coexistence. PMID- 14522175 TI - Incomplete mixing promotes species coexistence in a lottery model with permanent spatial heterogeneity. AB - For many marine organisms, the population dynamics in multiple habitats are affected by migration of planktonic larvae. We herein examine the effect of incomplete larval mixing on the condition for species coexistence. The system consists of two heterogeneous habitats, each composed of a number of sites occupied by adults of two species. Larvae produced in a habitat form a pool and migrate to the pool of the other habitat. When an adult dies, the vacant site becomes occupied by an individual randomly chosen from the larval pool. We study (1). the invasibility of a inferior species which has no advantage in either habitats, (2). the dynamics when larval migration and competition among adults are symmetric between habitats, and (3). the case with unidirectional migration. The coexistence of competitors is more likely to occur when larval migration is weak. PMID- 14522176 TI - Food web dynamics in correlated and autocorrelated environments. AB - The densities of populations in a community or food web vary as a consequence of both population interactions and environmental (e.g. weather) fluctuations. Populations often respond to the same kinds of environmental fluctuations, and therefore experience correlated environments. Furthermore, some environmental factors change slowly over time, thereby producing positive environmental autocorrelation. We show that the effects of environmental correlation and autocorrelation on the dynamics of the populations in a food web can be large and unintuitive, but can be understood by analyzing the eigenvectors of the community (system) matrix of interactions among populations. For example, environmental correlation and autocorrelation may either obscure or enhance the cyclic dynamics that generally characterize predator-prey interactions even when there is no direct effect of the environment on how species interact. Thus, understanding the population dynamics of species in a food web requires explicit attention to the correlation structure of environmental factors affecting all species. PMID- 14522177 TI - Quasicycles revisited: apparent sensitivity to initial conditions. AB - Environmental noise is known to sustain cycles by perturbing a deterministic approach to equilibrium that is itself oscillatory. Quasicycles produced in this way display a regular period but varied amplitude. They were proposed by Nisbet and Gurney (Nature 263 (1976) 319) as one possible explanation for population fluctuations in nature. Here, we revisit quasicyclic dynamics from the perspective of nonlinear time series analysis. Time series are generated with a predator-prey model whose prey's growth rate is driven by environmental noise. A method for the analysis of short and noisy data provides evidence for sensitivity to initial conditions, with a global Lyapunov exponent often close to zero characteristic of populations 'at the edge of chaos'. Results with methods restricted to long time series are consistent with a finite-dimensional attractor on which dynamics are sensitive to initial conditions. These results are compared with those previously obtained for quasicycles in an individual-based model with heterogeneous spatial distributions. Patterns of sensitivity to initial conditions are shown to differentiate phase-forgetting from phase-remembering quasicycles involving a periodic driver. The previously reported mode at zero of Lyapunov exponents in field and laboratory populations may reflect, in part, quasicyclic dynamics. PMID- 14522178 TI - From basic research to product development: not an easy way. PMID- 14522179 TI - Isolation of polymorphic microsatellite loci from the tapeworm Echinococcus multilocularis. AB - Two microsatellites were isolated from a genomic library of Echinococcus multilocularis. The microsatellites, designated EMms1 and EMms2, consist of tandem repeats of CAC-trinucleotide unit. Southern blot hybridization suggests that each of them is a single locus. Using fox-derived wild tapeworms (N=104), PCR-amplification of microsatellites was performed to assess the usefulness of these loci. We found four alleles of EMms1 and two alleles of EMms2. The heterozygosities observed were 10.6% in EMms1 and 7.7% in EMms2. The result suggests that both selfing and outcrossing occur in the adult stage of E. multilocularis. PMID- 14522180 TI - Population genetic structure and cladistic analysis of Trypanosoma brucei isolates. AB - Using a novel multilocus DNA marker analysis method, we studied the population genetic structure of Trypansoma brucei stocks and derived clones isolated from animal and rhodesiense sleeping sickness patients during a national sleeping sickness control program in Mukono district, Uganda. We then performed a cladistic analysis to trace relationships and evolution, using stocks and clones recovered from geographically and temporally matched hosts, including inter strain comparisons with T. b. gambiense stocks and clones. Our results show that while there was close genetic relatedness among parasite populations from the same geographical region, micro-heterogeneities exist between different stocks. Data are presented that indicate that not every human sleeping sickness focus may be associated with a particular human-infective trypanosome strain responsible for long-term stability of the reference focus. We provide evidence of genetic sub-structuring among type 1 T. b. gambiense stocks, which has potentially important implications for molecular epidemiology of T. brucei. PMID- 14522181 TI - Occurrence of recombinant genotypes of Anisakis simplex s.s. and Anisakis pegreffii (Nematoda: Anisakidae) in an area of sympatry. AB - The anisakid nematode populations collected from fish and stranded cetaceans along from Iberian Peninsula waters were morphologically identified as corresponding to the Anisakis simplex complex. In order to realise their molecular identification and to analyse the extent of genetic variation, the entire ITS (ITS1, 5.8S rDNA gene and ITS2) and the mitochondrial small subunit of rRNA were pcr-amplified and sequenced. Digestions of the amplified its region with HinfI and HhaI allowed the identification of three different genotypes, belonging to A. simplex s.s., A. pegreffii and a yet not described recombinant genotype. The ITS sequences of the recombinant genotypes showed the presence of heterozygotes C/T at position 240 and 256 of the aligned sequence. Otherwise, the analysis of mtDNA sequences showed the existence of a different parental origin for recombinant genotypes. In order to check if they can be the products of a polymorphism normally occurring both in A. pegreffii and in A. simplex s.s., and/or the existence of an incomplete concerted evolution, three samples were also collected as controls in isolated geographic areas, where sympatric coexistence between A. simplex s.s. and A. pegreffii does not occur. The results supports the hypothesis that the recombinant individuals may be a product of interspecific hybridisation, and describe the Iberian Peninsula waters as a hybrid zone for the two sibling species. PMID- 14522182 TI - Clinical and genetic risk factors for the development of multi-drug resistant tuberculosis in non-HIV infected patients at a tertiary care center in India: a case-control study. AB - Genetic susceptibility of the host to multi-drug resistant tuberculosis (MDR-TB) is not fully understood. We undertook a case-control study at a tertiary care center at New Delhi, India to identify the clinical and genetic predictors of MDR TB as compared to the drug sensitive TB cases. Patients with multi-drug resistant tuberculosis were identified on the basis of drug sensitivity testing by the proportion method. Treatment was initiated according to standard norms and all patients were followed up during the period. Genomic DNA extracted from the peripheral blood mononuclear cell pellet was used for amplification of HLA class II region (second exon) with a set of forward (5') and reverse (3') primers. A sequence specific 5' biotinylated probes were used to determine 12 DRB1, 8 DQA1 and 13 DQB1 alleles by the PCR-SSOP method. Past history of disease, higher severity of illness, inadequacy of drug treatment and presence of HLA-DRB1*14, DQB1*0503 and DQB1*0502 alleles were found to be significant risk factors for MDR TB. Multivariate analysis identified poor past compliance to treatment (odds ratio, OR=6.6; 95% confidence interval, CI [2.0-21.5]), higher number of cavities (OR=6; 95% CI [2.1-17.3]) in chest radiographs and the presence of the HLA DRB1*14 allele (OR=8.2; 95% CI [2.1-31.3]) as independent predictors of MDR-TB. Our results suggest that a combination of clinical and immunogenetic parameters could provide better information on drug resistance in tuberculosis with implications in therapy. PMID- 14522183 TI - Origin, world-wide dispersion, bio-geographical diversification, radiation and recombination: an evolutionary history of Yam mild mosaic virus (YMMV). AB - We developed an evolutionary epidemiological approach to understand the regional and world-wide dispersion of Yam mild mosaic virus (YMMV) by retracing its evolutionary history. Analyses of the distribution and the prevalence of YMMV in the Caribbean islands of Guadeloupe and Martinique, and in French Guyana revealed that YMMV has a wide repartition and different prevalence on Dioscorea alata L. (Asian and Oceanic origin), on D. cayenensis Lam.-D. rotundata Poir. (African origin) and on D. trifida L. (Amazon and the Caribbean origin) in this region. Considering the data on the current dispersion of the virus and the evolution and the history of the yams, the phylogenetic analysis of the 3' terminal part of the YMMV genome gave a consistent support of the Asian-Pacific origin of YMMV from D. alata species. The YMMV phylogenetic tree is star-like, suggesting an early split of the genetic lineages. An important part of the clades is constituted by a single lineage arisen by recombination. The largest emerging monophyletic group illustrates well YMMV geographical dispersion. This evolutionary pattern contrasts with the one revealed by the African distinct lineages and by the second significant monophyletic group, for which a host adaptation to D. trifida is suggested. The analysis of the pattern of nucleotide substitutions in the CP gene revealed that purifying selection dominates the evolution of the CP of potyviruses and strongly operates on the YMMV. Switching events, radiation, host and geographical adaptation and recombination events are proposed as major traits of YMMV evolutionary history. PMID- 14522184 TI - Multilocus genotyping of Cryptosporidium parvum Type 2: population genetics and sub-structuring. AB - Cryptosporidium parvum is an intracellular protozoan parasite that infects the gastrointestinal tract of humans and other mammals. It has significant economic importance as a pathogen of livestock and, as there is no effective treatment or vaccine available, understanding transmission routes and identifying sources of infection is key to preventing future outbreaks and controlling this disease. In this study we have determined the multilocus genotype (MLG) of 240 C. parvum Type 2 (bovine) isolates using a combination of seven micro- and minisatellite markers. These isolates were collected over a period of 19 months and are from three different geographical locations within Scotland and three different host species. The results of this study have enabled us to address questions concerning C. parvum population genetics in relation to host, temporal and geographical sub-structuring. We identified 48 multilocus genotypes within the Type 2 C. parvum isolates and found no evidence to support geographic or temporal sub-structuring of the populations. However host sub-structuring was identified within the human Type 2 population highlighting the potential use of such a typing system in understanding the epidemiology of this parasite in addition to raising interesting questions with regard to its population genetic structure. We also isolated two C. parvum 'monkey type' isolates from two separate human cases indicating that this genotype is not restricted to monkey hosts with the multilocus genotypes of these isolates distinguishing them from all other isolates. PMID- 14522185 TI - SARS associated coronavirus has a recombinant polymerase and coronaviruses have a history of host-shifting. AB - The sudden appearance and potential lethality of severe acute respiratory syndrome associated coronavirus (SARS-CoV) in humans has focused attention on understanding its origins. Here, we assess phylogenetic relationships for the SARS-CoV lineage as well as the history of host-species shifts for SARS-CoV and other coronaviruses. We used a Bayesian phylogenetic inference approach with sliding window analyses of three SARS-CoV proteins: RNA dependent RNA polymerase (RDRP), nucleocapsid (N) and spike (S). Conservation of RDRP allowed us to use a set of Arteriviridae taxa to root the Coronaviridae phylogeny. We found strong evidence for a recombination breakpoint within SARS-CoV RDRP, based on different, well supported trees for a 5' fragment (supporting SARS-CoV as sister to a clade including all other coronaviruses) and a 3' fragment (supporting SARS-CoV as sister to group three avian coronaviruses). These different topologies are statistically significant: the optimal 5' tree could be rejected for the 3' region, and the optimal 3' tree could be rejected for the 5' region. We did not find statistical evidence for recombination in analyses of N and S, as there is little signal to differentiate among alternative trees. Comparison of phylogenetic trees for 11 known host-species and 36 coronaviruses, representing coronavirus groups 1-3 and SARS-CoV, based on N showed statistical incongruence indicating multiple host-species shifts for coronaviruses. Inference of host species associations is highly sensitive to sampling and must be considered cautiously. However, current sampling suggests host-species shifts between mouse and rat, chicken and turkey, mammals and manx shearwater, and humans and other mammals. The sister relationship between avian coronaviruses and the 3' RDRP fragment of SARS-CoV suggests an additional host-species shift. Demonstration of recombination in the SARS-CoV lineage indicates its potential for rapid unpredictable change, a potentially important challenge for public health management and for drug and vaccine development. PMID- 14522187 TI - Characterization of municipal solid waste combustion in a grate furnace. AB - The objective of this paper is to evaluate the combustion process of municipal solid waste combustion in a grate furnace both experimentally and numerically by using data of a reference experiment with over-stoichiometric primary air supply. Measurements were carried out inside the combustion chamber of a pilot plant by monitoring temperatures and sampling gaseous combustion products along the bed surface. The data were assessed using elemental and energy balances. Experimental data of the axial temperature profiles of the flue gas, the fuel bed and the grate bars, as well as local gas flows and the flue gas composition measured above the fuel bed along the grate were used to describe the conversion process, including drying and carbon burnout. These data served as input to model the thermo- and fluid dynamic processes of the gas phase above the bed inside the combustion chamber. For this purpose the commercial code FLUENT was employed to carry out the simulations. Thus, the turbulent temperature, flow and species distributions in the combustion chamber of the pilot waste incinerator TAMARA were predicted. The results of the FLUENT modeling showed that under the prevailing conditions the flue gas burnout is almost completed before entering the first flue due to high temperatures, effective mixing and sufficient residence times of the flue gas inside the combustion chamber. This agrees well with the experimental results inside the first flue. On the basis of the above mentioned results, design and parametric studies can be carried out in a more efficient way by saving cost and time. PMID- 14522188 TI - Recycling of spent nickel-cadmium batteries based on bioleaching process. AB - Only 1-2 percent of discarded dry batteries are recovered in China. It is necessary to find an economic and environmentally friendly process to recycle dry batteries in this developing country. Bioleaching is one of the few techniques applicable for the recovery of the toxic metals from hazardous spent batteries. Its principle is the microbial production of sulphuric acid and simultaneous leaching of metals. In this study, a system consisting of a bioreactor, settling tank and leaching reactor was developed to leach metals from nickel-cadmium batteries. Indigenous thiobacilli, proliferated by using nutritive elements in sewage sludge and elemental sulphur as substrates, was employed in the bioreactor to produce sulphuric acid. The overflow from the bioreactor was conducted into the settling tank. The supernatant in the settling tank was conducted into the leaching reactor, which contained the anode and cathodic electrodes obtained from nickel-cadmium batteries. The results showed that this system was valid to leach metals from nickel-cadmium batteries, and that the sludge drained from the bottom of the settling tank could satisfy the requirements of environmental protection agencies regarding agricultural use. PMID- 14522189 TI - Technical assessment of fuel cell operation on anaerobic digester gas at the Yonkers, NY, wastewater treatment plant. AB - This paper summarizes the results of a 2-year field test to assess the performance of a specially modified commercial phosphoric acid 200-kW fuel cell power plant to recover energy from anaerobic digester gas (ADG) which has been cleansed of contaminants (sulfur and halide compounds) using a patented gas pretreatment unit (GPU). Specific project goals include characterization of the fuel cell power plant emissions and verification of the GPU performance for removing sulfur contaminants. To remove halide contaminants from the ADG, a halide guard, consisting of a vessel with a metal oxide supported on alumina, was incorporated into the fuel cell reactant supply. This first-of-a-kind demonstration was conducted at the Yonkers, NY, wastewater treatment plant, a sewage processing facility owned and operated by Westchester County. Results have demonstrated that the ADG fuel cell power plant can produce electrical output levels close to full power (200 kW) with negligible air emissions of CO, NO(x), and SO(2). The GPU removed virtually 100% of H(2)S and 88% of organic sulfur, bringing the overall sulfur removal efficiency of the GPU to over 99%. The halide guard removed up to 96% of the halides exiting the GPU. PMID- 14522190 TI - Environmental impact of highway construction and repair materials on surface and ground waters. Case study: crumb rubber asphalt concrete. AB - The practice of incorporating certain waste products into highway construction and repair materials (CRMs) has become more popular. These practices have prompted the National Academy of Science, National Cooperative Highway Research Program (NCHRP) to research the possible impacts of these CRMs on the quality of surface and ground waters. State department of transportations (DOTs) are currently experimenting with use of ground tire rubber ( crumb rubber) in bituminous construction and as a crack sealer. Crumb rubber asphalt concrete (CR AC) leachates contain a mixture of organic and metallic contaminants. Benzothiazole and 2(3H)-benzothiazolone (organic compounds used in tire rubber manufacturing) and the metals mercury and aluminum were leached in potentially harmful concentrations (exceeding toxic concentrations for aquatic toxicity tests). CR-AC leachate exhibited moderate to high toxicity for algae ( Selenastrum capriconutum) and moderate toxicity for water fleas ( Daphnia magna). Benzothiazole was readily removed from CR-AC leachate by the environmental processes of soil sorption, volatilization, and biodegradation. Metals, which do not volatilize or photochemically or biologically degrade, were removed from the leachate by soil sorption. Contaminants from CR-AC leachates are thus degraded or retarded in their transport through nearby soils and ground waters. PMID- 14522191 TI - Correlations between homologue concentrations of PCDD/Fs and toxic equivalency values in laboratory-, package boiler-, and field-scale incinerators. AB - The toxic equivalency (TEQ) values of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) are predicted with a model based on the homologue concentrations measured from a laboratory-scale reactor (124 data points), a package boiler (61 data points), and operating municipal waste incinerators (114 data points). Regardless of the three scales and types of equipment, the different temperature profiles, sampling emissions and/or solids (fly ash), and the various chemical and physical properties of the fuels, all the PCDF plots showed highly linear correlations (R(2)>0.99). The fitting lines of the reactor and the boiler data were almost linear with slope of unity, whereas the slope of the municipal waste incinerator data was 0.86, which is caused by higher predicted values for samples with high measured TEQ. The strong correlation also implies that each of the 10 toxic PCDF congeners has a constant concentration relative to its respective total homologue concentration despite a wide range of facility types and combustion conditions. The PCDD plots showed significant scatter and poor linearity, which implies that the relative concentration of PCDD TEQ congeners is more sensitive to variations in reaction conditions than that of the PCDF congeners. PMID- 14522192 TI - Permanganate oxidation of sorbed polycyclic aromatic hydrocarbons. AB - The polycyclic aromatic hydrocarbons (PAH) that contaminate soils at many industrial and government sites are resistant to natural biotic and abiotic degradation processes. The recalcitrant nature of these compounds may require aggressive chemical treatment to effectively remediate these sites. This study was conducted to assess the viability of permanganate oxidative treatment as a method to reduce PAH concentration in contaminated soils. Study results demonstrated a reduction in soil sorbed concentration for a mixture of six PAHs that included anthracene, benzo(a)pyrene, chrysene, fluoranthene, phenanthrene, and pyrene by potassium permanganate (KMnO4) oxidative treatment. The greatest reduction in soil concentration was observed for benzo(a)pyrene, pyrene, phenanthrene, and anthracene at 72.1, 64.2, 56.2, and 53.8%, respectively, in 30 min at a KMnO4 concentration of 160 mM. Minimal reductions in fluoranthene and chrysene concentration were observed at 13.4 and 7.8%, respectively, under the same conditions. A relative chemical reactivity order of benzo(a)pyrene>pyrene>phenanthrene>anthracene>fluoranthene>chrysene towards permanganate ion was observed. Aromatic sextet theory was applied to the degradation results to explain the highly variable and compound-specific chemical reactivity order. PMID- 14522193 TI - An investigation of the presence of methane and other gases at the Uzundere-Izmir solid waste disposal site, Izmir, Turkey. AB - Izmir is a large metropolitan city with a population of 3,114,860. The city consists of 27 townships, each township has a population of not less than 10,000 inhabitants. The two major solid waste disposal sites are in the townships of Uzundere and Harmandali. The amount of solid waste that is disposed at each of these sites is about 800 and 1800 t/day, respectively. In Uzundere, compost is produced from the organic fraction of urban solid wastes while the residual material is deposited at a disposal site with a remaining capacity of 700,000 m(3) as of 2001. Gas monitoring and measurements were carried out at the disposal site in Uzundere. For this purpose, nine sampling wells were drilled on selected locations. Each well was furnished with perforated metal pipes suitable for gas monitoring and measurements. The following gases were monitored: O(2), CH(4), CO, CO(2), and H(2)S. The most important finding was that the concentrations of CH(4) in the wells ranged from 7 to 57%. Dilution of the CH(4) by O(2) down to the LEL levels (5-15%) is always possible and poses a continuing risk at the site. Furthermore, the levels of O(2) require that access to the site be limited to only authorized personnel. PMID- 14522194 TI - Building waste management in Bulgaria: challenges and opportunities. AB - Building waste recycling as aggregates is a modern approach for preventing environmental pollution through both reducing the stocks of waste and decreasing the use of natural aggregates. The reuse of building waste is a relatively new issue for Bulgaria despite the existing considerable quantity of building waste and the significant changes in the environmental rules applied. The paper discusses generated and potential waste streams in Bulgaria in the context of the social and economic restructuring and recent urban development undergone by the country. The main preliminary conditions for developing the recycling activity such as: streams of building waste, experience in recycling, technical and environmental standardization, appropriate technologies, etc. are examined. The authors analyze current practice and research activities with regard to the implementation of advanced EU building-waste recycling methods. Conclusions are drawn about existing opportunities and the priorities of the needed building waste management strategy in the country. PMID- 14522195 TI - Processing of copper anodic-slimes for extraction of valuable metals. AB - This work focuses on processing of anodic slimes obtained from an Egyptian copper electrorefining plant. The anodic slimes are characterized by high concentrations of copper, lead, tin and silver. The proposed hydrometallurgical process consists of two leaching stages for the extraction of copper (H(2)SO(4)-O(2)) and silver (thiourea-Fe3+), and pyrometallurgical treatment of the remaining slimes for production of Pb-Sn soldering alloy. Factors affecting both the leaching and smelting stages were studied. PMID- 14522196 TI - Environmental evaluation of different variants of the chromium compound production model using chromic waste. AB - This work presents research results on the evaluation of environmental effects of utilizing chromic waste in chromium compound production processes. The comprehensive evaluation of three chromium compound production models takes into consideration the total cumulated hazard coefficients. The implementation of the new chromium compound production model into industrial practice in 1999 allows the hazard to the natural environment to decrease by 75% in relation to the hazard caused by the old production model used until 1995. The coefficient of the target chromium compound production model was 199%, much higher than 100%. This results from the negative values of the total cumulated hazard coefficients for those cases where the sodium chromate production unit could be a big "consumer" of chromic waste. PMID- 14522197 TI - Role of aluminous component of fly ash on the durability of Portland cement-fly ash pastes in marine environment. AB - The durability, of mixtures of two kinds of Spanish fly ashes from coal combustion (ASTM class F) with 0, 15 and 35% replacement of Portland cement by fly ash, in a simulated marine environment (Na(2)SO(4)+NaCl solution of equivalent concentration to that of sea water: 0.03 and 0.45 M for sulphate and chloride, respectively), has been studied for a period of 90 days. The resistance of the different mixtures to the attack was evaluated by means of the Koch Steinegger test. The results showed that all the mixtures were resistant, in spite of the great amount of Al(2)O(3) content of the fly ash. The diffusion of SO(4)(2-), Na+ and Cl- ions through the pore solution activated the pozzolanic reactivity of the fly ashes causing the corresponding microstructure changes, which were characterized by X-ray diffraction (XRD), mercury intrusion porosimetry (MIP) and scanning electron microscopy (SEM). As a result, the flexural strength of the mixtures increased, principally for the fly ash of a lower particle size and 35% of addition. PMID- 14522199 TI - Three-dimensional visualization and morphometry of small airways from microfocal X-ray computed tomography. AB - Physiological morphometry is a critical factor in the flow dynamics in small airways. In this study, we visualized and analyzed the three-dimensional structure of the small airways without dehydration and fixation. We developed a two-step method to visualize small airways in detail by staining the lung tissue with a radiopaque solution and then visualizing the tissue with a cone-beam microfocal X-ray computed tomographic (CT) system. To verify the applicability of this staining and CT imaging (SCT) method, we used the method to visualize small airways in excised rat lungs. By using the SCT method to obtain continuous CT images, three-dimensional branching and merging bronchi ranging from 500 to 150 microm (the airway generation=8-16) were successfully reconstructed. The morphometry of the small airways (diameter, length, branching angle and gravity angle between the gravity direction and airway vector) was analyzed using the three-dimensional thinning algorithm. The diameter and length exponentially decreased with the airway generation. The asymmetry of the bifurcation decreased with generation and one branching angle decided the other pair branching angle. The SCT method is the first reported method that yields faithful high-resolution images of soft tissue geometry without fixation and the three-dimensional morphometry of small airways is useful for studying the biomechanical dynamics in small airways. PMID- 14522200 TI - A potential role of smooth muscle tone in early hypertension: a theoretical study. AB - A conspicuous long-term consequence of hypertension is a thickening of the arterial wall, which many suggest returns the circumferential wall stress toward its normal value. This thickening results from an increase in smooth muscle and extracellular matrix, with the associated growth and remodeling processes depending on a host of regulatory signals that likely include the altered mechanical environment. Although the precise mechanotransduction pathways remain unknown, we propose that vasoconstriction may be an early response of the arterial wall to a step-change in pressure. In particular, computations suggest that such a response can decrease the magnitude and transmural gradients of the pressure-induced wall stresses and return the mean wall shear stress toward its homeostatic value. Such an initial 'compensatory vasoconstriction' could also help set into motion subsequent growth and remodeling responses due to growth regulatory characteristics of the vasoactive molecules (e.g., nitric oxide, endothelin-1, angiotensin-II). Although the consequences of growth and remodeling have been the focus of prior biomechanical and histological studies, early responses dictate subsequent developments and therefore deserve increased attention in vascular biomechanics and mechanobiology. PMID- 14522201 TI - Comparative in vitro study on the long term performance of cemented hip stems: validation of a protocol to discriminate between "good" and "bad" designs. AB - The long-term clinical success of cemented hip stems is influenced both by the implant design, and by the surgical procedure. A methodology is proposed for discriminating between implant designs with different clinical outcomes. The protocol was designed with industrial pre-clinical validation in mind. Two cemented stem types were tested, one (Lubinus SPII) having good and the other (Muller Curved) having poor clinical outcomes. Three implants for each type were subjected to a mechanical in vitro test of one million loading cycles. Each cycle reproduced the load components of stair climbing. Interface shear micromotion was measured during the test in the direction of rotation and along the stem axis. The stem roughness before and after the test was compared. After the test, the cement mantles were retrieved and inspected through dye penetrants to detect evidences of micro-damage. For each specimen, the events of the loosening process were examined, based on the in vitro data available, so as to analyze the whole failure mechanism. The protocol developed was sensitive to the implant design, with significantly different results being found for the two stem types, both in terms of stem-cement micromotions, surface roughness alteration, and cement mantle damage. The information yielded by the three different investigation techniques was consistent for each of the two groups of specimens tested, allowing a better understanding of the failure process. In vitro inducible micromotion and permanent migration measurements, together with cement-stem interface fretting damage and cement fatigue damage, can help predicting the clinical performance of cemented stems. PMID- 14522202 TI - Heelstrike and the pathomechanics of osteoarthrosis: a simulation study. AB - A simulation model and trajectory matching method were developed to investigate the differences between two swing phase gait patterns; one giving rise to a large impulse at heelstrike, the other giving a small impulse. Subtle changes in the kinematics of the swing leg dramatically reduced the vertical contact velocity of the ankle at the moment of heelstrike. Phasing of the hip flexor muscles at the start of swing was responsible for the level of impulse observed at heelstrike. PMID- 14522203 TI - Heelstrike and the pathomechanics of osteoarthrosis: a pilot gait study. AB - Involvement of mechanical factors in osteoarthrosis (OA) has been well documented. For OA of the human lower limb, the impulse imparted at heelstrike has been suggested as a pathogenic factor. It has also been reported that there is a large amount of variation in the level of impulse experienced by different individuals, and it is suggested that those who experience large impulses are at a greater risk of developing OA. The current study investigated gait patterns of 12 normal subjects to establish the gait determinants responsible for producing large impulses at heelstrike. The results suggest that subtle variations in the early part of the swing phase pattern are responsible for large differences in the impulse experienced at heelstrike; the usually reported gait variables mask these variations. PMID- 14522204 TI - Improvement of the reliability of ceramic hip joint implants. AB - The aim of this article is to present the optimization of a proof test procedure of ceramic hip joint ball heads. The proof test rejects defective samples in the production line before being implanted into human body. Thereby on every ceramic ball head a static load is applied, which is somewhat higher than the maximum physiological load. The magnitude of the applied load should not damage the samples which are free of flaws in the high stress area. The configuration of the proof test influences the stress distribution in the ball head, which should be similar to the physiological case. To determine the stress distribution, a non linear finite element (FE) analysis was performed and the results were validated by measurements. With an iterative approach based on FE calculations the proof test configuration was optimized in such a way that the stress distribution in the ball head is similar to the stress distribution in vivo. In this study all ball heads showed very high fatigue resistance after being proof tested and fulfilled the requirements of the FDA (Food and Drug Administration, USA) described in the Guidance Document for the Preparation of Premarket Notifications for Ceramic Ball Hip System. The probability of a fracture of an implanted ceramic ball head can be decreased by the presented optimized proof test procedure. Latter can thus improve the reliability of ceramic hip joint ball heads. The study was supported by the KTI (Commission for Technology and Innovation, Switzerland). PMID- 14522205 TI - In vitro evaluation of combined graft deformation in anterior cruciate ligament reconstructions. AB - In this paper, we present a simple method of calculating deformation of an anterior cruciate ligament graft in combined elongation, bending and total twisting. We also report our results on these deformations for three types of ligament reconstruction in cadaver knees: two isometric reconstructions, using either a biological bone-patellar tendon-bone autograft or an artificial Trevira prosthesis, and modified over-the-top reconstruction with the prosthesis. The data show that the modified OTT technique produced elongation of the graft equivalent to that of the synthetic isometric technique, but significantly less than isometric reconstruction with the biological graft (p<0.05, ANOVA). Moreover, the grafts were subjected to bending and twisting. PMID- 14522206 TI - Mechanics of the tapered interference fit in dental implants. AB - In evaluation of the long-term success of a dental implant, the reliability and the stability of the implant-abutment interface plays a great role. Tapered interference fits provide a reliable connection method between the abutment and the implant. In this work, the mechanics of the tapered interference fits were analyzed using a closed-form formula and the finite element (FE) method. An analytical solution, which is used to predict the contact pressure in a straight interference, was modified to predict the contact pressure in the tapered implant abutment interface. Elastic-plastic FE analysis was used to simulate the implant and abutment material behavior. The validity and the applicability of the analytical solution were investigated by comparisons with the FE model for a range of problem parameters. It was shown that the analytical solution could be used to determine the pull-out force and loosening-torque with 5-10% error. Detailed analysis of the stress distribution due to tapered interference fit, in a commercially available, abutment-implant system was carried out. This analysis shows that plastic deformation in the implant limits the increase in the pull-out force that would have been otherwise predicted by higher interference values. PMID- 14522207 TI - Experimental evaluation of an elastic foundation model to predict contact pressures in knee replacements. AB - Computational wear prediction is an attractive concept for evaluating new total knee replacement designs prior to physical testing and implementation. An important hurdle to such technology is the lack of in vivo contact pressure predictions. To address this issue, this study evaluates a computationally efficient simulation approach that combines the advantages of rigid and deformable body modeling. The hybrid method uses rigid body dynamics to predict body positions and orientations and elastic foundation theory to predict contact pressures between general three-dimensional surfaces. To evaluate the method, we performed static pressure experiments with a commercial knee implant in neutral alignment using flexion angles of 0, 30, 60, and 90 degrees and loads of 750, 1500, 2250, and 3000N. Using manufacturer CAD geometry for the same implant, an elastic foundation model with linear or nonlinear polyethylene material properties was implemented within a commercial multibody dynamics software program. The model's ability to predict experimental peak and average contact pressures simultaneously was evaluated by performing dynamic simulations to find the static configuration. Both the linear and nonlinear material models predicted the average contact pressure data well, while only the linear material model could simultaneously predict the trends in the peak contact pressure data. This novel modeling approach is sufficiently fast and accurate to be used in design sensitivity and optimization studies of knee implant mechanics and ultimately wear. PMID- 14522208 TI - Development and testing of a modular strain measurement clip. AB - A novel, multi-use, low-stiffness and low-cost transducer for measuring in vitro strains has been developed and tested. Currently available strain measurement methods are either too expensive, too complicated or too inflexible for multi-use strain measurement. The stainless-steel modular strain measurement clip introduced here was instrumented with four 350 Omega axial strain gauges in a full Wheatstone bridge configuration to take advantage of commonly available strain gauge amplifier equipment. Adjustable extension arms were designed to allow greater application versatility. The clip was calibrated and produced a linear response (R(2)>0.99) over a minimum of 1.04 mm at high amplifier gain. With reduced amplifier settings, testing showed a linear response over a range of 30.5 mm (R(2)>0.99). Clip stiffness was 0.6N/mm of extension arm tip displacement for minimal instrumentation artifact. A validation test was conducted through a comparison of strain clips, surface-mounted strain gauges and theoretical strain in an aluminium rod subjected to axial tensile loading. The two measurement techniques were used to determine the modulus of elasticity of the aluminium rod. Results were within 6% of the known modulus of elasticity for aluminium. A comparative biomechanical test was also performed on an equine third metacarpal specimen. The traditional bonded strain gauging method produced similar results as the new strain clip, but failed to measure ultimate strains since all strain gauges failed prior to specimen failure. Further investigations into the multiple uses of the clip are underway and recommendations for future versions of the clip are given. PMID- 14522209 TI - Longer moment arm results in smaller joint moment development, power and work outputs in fast motions. AB - Effects of moment arm length on kinetic outputs of a musculoskeletal system (muscle force development, joint moment development, joint power output and joint work output) were evaluated using computer simulation. A skeletal system of the human ankle joint was constructed: a lower leg segment and a foot segment were connected with a hinge joint. A Hill-type model of the musculus soleus (m. soleus), consisting of a contractile element and a series elastic element, was attached to the skeletal system. The model of the m. soleus was maximally activated, while the ankle joint was plantarflexed/dorsiflexed at a variation of constant angular velocities, simulating isokinetic exercises on a muscle testing machine. Profiles of the kinetic outputs (muscle force development, joint moment development, joint power output and joint work output) were obtained. Thereafter, the location of the insertion of the m. soleus was shifted toward the dorsal/ventral direction by 1cm, which had an effect of lengthening/shortening the moment arm length, respectively. The kinetic outputs of the musculoskeletal system during the simulated isokinetic exercises were evaluated with these longer/shorter moment arm lengths. It was found that longer moment arm resulted in smaller joint moment development, smaller joint power output and smaller joint work output in the larger plantarflexion angular velocity region (>120 degrees/s). This is because larger muscle shortening velocity was required with longer moment arm to achieve a certain joint angular velocity. Larger muscle shortening velocity resulted in smaller muscle force development because of the force-velocity relation of the muscle. It was suggested that this phenomenon should be taken into consideration when investigating the joint moment-joint angle and/or joint moment-joint angular velocity characteristics of experimental data. PMID- 14522210 TI - Tensile damage and its effects on cortical bone. AB - Plexiform bovine bone samples are repeatedly loaded in tension along their longitudinal axis. In order to induce damage in the bone tissue, bone samples are loaded past their yield point. Half of the bone samples from the damaged group were stored in saline to allow for viscoelastic recovery while the others were decalcified. Tensile tests were conducted on these samples to characterize the effects of damage on the mechanical behavior of the organic matrix (decalcified samples) as well as on bone tissue (stored in saline). The ultimate strain of the damaged decalcified bone is 29% higher compared to that of non-damaged decalcified (control) bone. The ultimate stresses as well as the elastic moduli are similar in both decalcified groups. This phenomenon is also observed in other collagenous tissue (tendon and ligament). This may suggest that damage in bone is caused by shear failure of the organic matrix; transverse separation of the collagen molecules or microfibrils from each other. In contrast, there is a trend towards lowered ultimate strains in damaged bone, which is soaked in saline, with respect to control bone samples (not damaged). The damaged bone tissue exhibits a bi-linear behavior in contrast to the mechanical behavior of non-damaged bone. The initial elastic modulus (below 55 MPa) and ultimate strength of damaged bone are similar to that in non-damaged bone. PMID- 14522211 TI - Inverse dynamic investigation of voluntary trunk movements in weightlessness: a new microgravity-specific strategy. AB - Present investigation faces the question of quantitative assessment of exchanged forces and torques at the restraints during whole body posture exercises in long term microgravity. Inverse dynamic modelling and total angular momentum at the ankle joint were used in order to reconstruct movement dynamics at the restraining point, represented by the ankle joint. The hypothesis is that the minimisation of the torques at the interface point assumes a key role in movement planning in 0 g. This hypothesis would respond to an optimisation of muscles activity, a minimisation of energy expenditure and therefore an accurate control of body movement. Results show that the 0 g movement strategy adopted ensures that the integral of the net ankle moment between the beginning and the end of the movement is zero. This expected mechanical constraint is not satisfied when 0 g movement dynamics is simulated using terrestrial kinematics. This accounts for a significant imposed change of movement strategy. Particularly, the efficient compensation of the inertial effects of the segments in terms of total angular momentum at the ankle joint was evidenced. These results explain the exaggerated axial synergies, observed on kinematics and which moved centre of mass (CM) backward from its already backward initial positioning, as a tool for enhancing the compensation and achieving the desired minimisation of the torques exchanges at the restraints. PMID- 14522212 TI - Biomechanical modeling and optimal control of human posture. AB - The present work describes the biomechanical modeling of human postural mechanics in the saggital plane and the use of optimal control to generate open-loop raising-up movements from a squatting position. The biomechanical model comprises 10 equivalent musculotendon actuators, based on a 40 muscles model, and three links (shank, thigh and HAT-Head, Arms and Trunk). Optimal control solutions are achieved through algorithms based on the Consistent Approximations Theory (Schwartz and Polak, 1996), where the continuous non-linear dynamics is represented in a discrete space by means of a Runge-Kutta integration and the control signals in a spline-coefficient functional space. This leads to non linear programming problems solved by a sequential quadratic programming (SQP) method. Due to the highly non-linear and unstable nature of the posture dynamics, numerical convergence is difficult, and specific strategies must be implemented in order to allow convergence. Results for control (muscular excitations) and angular trajectories are shown using two final simulation times, as well as specific control strategies are discussed. PMID- 14522213 TI - The shape of the force-elbow angle relationship for maximal voluntary contractions and sub-maximal electrically induced contractions in human elbow flexors. AB - The force-length relationship is a basic property of skeletal muscle. Knowledge of this relationship is necessary for most analyses of human movement, and in simulation models predicting movement control strategies. Studies on animal muscles have shown that force-length relationships for sub-maximal contractions are not related through a simple scaling procedure to the relationship for maximal contractions. Furthermore, potentiation might produce a shift of sub maximal relative to maximal force-length relationships. In this study, we tested the hypothesis that human force-elbow angle relationships for sub-maximal unpotentiated contractions are shifted to larger elbow angles (i.e. larger muscle lengths) compared to the relationship for maximal voluntary contractions (MVC), and that this shift is reduced, or even abolished, for sub-maximal potentiated contractions. Force-elbow angle relationships (48-160 degrees) were obtained from healthy subjects (n=13). At each of nine tested elbow angles, the test set consisted of a single twitch (ST(pre)) and a doublet twitch (DT(pre)) stimulation of m. biceps brachii, followed by an MVC, followed by another single twitch (ST(post)) and a doublet twitch (DT(post)) stimulation. The single and doublet twitches induced sub-maximal contractions. The force-elbow angle relationships for the pre-MVC (unpotentiated) twitch contractions were shifted to larger angles compared to those obtained for MVC. The force-elbow angle relationships for the post-MVC (potentiated) twitch contractions were shifted to smaller angles compared to those obtained for the unpotentiated twitch contractions. These results support the idea that the shift to larger muscle lengths for the sub maximal, unpotentiated force-length relationships relative to the relationship for maximal contractions may be caused by a length-dependent Ca(2+) sensitivity that may be offset, at least in part, by potentiation. PMID- 14522214 TI - Force track analysis of contemporary hip simulators. AB - In an earlier paper, the authors presented the first verified method of computation of slide tracks in the relative motion between femoral head and acetabular cup of total hip prostheses. The method was applied for gait and for two hip simulator designs, and in a subsequent paper, for another eight designs. In the present paper, the track drawn by the resultant contact force, the so called force track, was studied in depth. The variations of sliding distance, sliding velocity and direction of sliding during a cycle, all of which are important with respect to wear, were computed for gait and for 11 hip simulator designs. Moreover, the product of the instantaneous load and increment of sliding distance was numerically integrated over a cycle. This integral makes it possible to compare clinical wear rates with those produced by hip simulators in terms of a wear factor. For the majority of contemporary hip simulators, the integral has so far been unknown. The computations revealed considerable differences, which are likely to explain the substantial differences in wear produced by the simulators. With the most common head diameter, 28 mm, the ranges for sliding distance per cycle, mean sliding velocity, total change of direction of sliding and integral were: 19.7-34.3 mm, 19.7-49.0 mm/s, 360-1513 degrees, and 17.4-43.5 Nm, respectively. PMID- 14522215 TI - On the elastic properties of arteries. AB - A new coefficient of elasticity is proposed that relates to the elastic state of the blood vessels. This measure is proposed as a result of the realization, from personal experience as well as from the international literature, of the difficulty in measuring the thickness of the blood vessels in vivo with acceptable precision. The measurement of E being dependent on the measurement of the thickness of the vessels becomes a highly unreliable proposition. Its relation to E (Young modulus) and to the pulse wave velocity (PWV) is established. We give three examples showing how the proposed coefficient can be measured. PMID- 14522216 TI - Folding-unfolding of FN-III domains in tenascin: an elastically coupled two-state system. AB - In a single-molecule atomic force microscopy (AFM) experiment, the tenascin molecule is stretched by an external force causing an elongation which is due to the unfolding of the FN-III modules. The features of the force-extension curves depend on the pulling speed and show a saw-tooth pattern (lower speeds) or a smooth pattern (higher speeds). In any case, the unfolded domains are elastically coupled to the unfolded modules, acting as transmitters of the external force. In this communication, the folding-unfolding process of the FN-III domains in tenascin is studied using reaction rate theory and a simple two-state model. The main hypothesis of the study is that, at microscopic level, the force needed to unfold a domain and the unfolding rate (unfolding velocity) can mimic the macroscopic process of measurement by AFM. As the external force is applied, the probability of unfolding increases as dictated by the reaction rate theory. Within this context, a relationship between the unfolding force and the unfolding velocity is obtained. The latter relation will describe microscopically the process in a phenomenological fashion. Moreover, while relating the results of this study with other experimental (AFM measurements) and theoretical (Monte Carlo simulations) data, we found that the graph of unfolding force-unfolding velocity is similar to that of external force-pulling velocity. The refolding process can also be studied within this model and the results show similar trends. The latter suggests a generic and universal behavior of such kind of molecular domains at least in the light of the proposed model. PMID- 14522217 TI - An approach for hip joint center calculation for use in seated postures. AB - In seated postures, such as those in office or automotive seats, locating the hip joint center (HJC) using three markers on the pelvis has been difficult if not impossible. A two-target approach by Bell et al. (J. Biomech. 23 (1990) 617) has been used, however, this method was shown to have inaccuracies when compared to the three-target method developed by Seidel et al. (J. Biomech. 28 (1995) 995). A new two-target method that is specific to the seated environment, has better accuracy than the Bell et al. approach, and is based on the Seidel et al. approach was developed and tested on 13 seated subjects. This new method used three targets and an initial reference file to estimate the HJC location. Once the HJC was located, assumptions were made that the magnitudes between the HJC and the respective anterior superior iliac spine, and the HJC and the respective lateral epicondyle remained constant. The primary concern when evaluating this new method was the affect of seated posture movement, in particular leg splay and spinal flexion on the assumptions. The results obtained with the new approach were compared to Seidel et al. and provided HJC locations with average differences of 3.8, 1.2 and 2.8mm for spinal flexion in the anterior/posterior, medial/lateral and superior/inferior directions, respectively, and 2.3, 1.0 and 1.4mm for knee splay. The proposed method provided better HJC estimation than the Bell et al. approach particularly in the superior/inferior dimensions. PMID- 14522218 TI - Finger force vectors in multi-finger prehension. AB - In a majority of studies on grasp, only normal forces were measured and only when a zero torque was exerted on a hand-held object. This study concerns finger force vectors during the torque production tasks. Subjects (n=8) stabilized a handle with an attachment that allowed for change of external torque from -1.5 to 1.5 Nm. Forces and moments exerted by the digit tips on the object were recorded. At the large (>-0.375 Nm) supination torques the index/middle and ring/little pairs of fingers generated oppositely directed tangential forces. The index and middle finger produced forces in a downward direction and therefore did not support the load. At a zero torque and pronation torques, the middle, ring and little fingers produced forces along nearly the same direction. The vector of the index finger force was always directed differently from the vectors of other finger forces, the angles ranged from 19 degrees 30' to 47 degrees 40'. The points of force application were systematically displaced with the torque, with the exception of the little finger. Tangential finger forces contributed substantially to the total torque exerted on the hand-held object. PMID- 14522219 TI - Locating women in law and psychiatry. PMID- 14522221 TI - Domestic violence: does the African context demand a different approach? PMID- 14522220 TI - False memory syndrome: undermining the credibility of complainants in sexual offences. PMID- 14522222 TI - Mothers who kill: cross-cultural patterns in and perspectives on contemporary maternal filicide. PMID- 14522223 TI - Manufacturing uncertainty: adverse effects of drug development for women. PMID- 14522225 TI - The impact of gender role ideology, male expectancies, and acculturation on wife abuse. PMID- 14522224 TI - Anorexia nervosa and personal identity: The accounts of patients and their parents. PMID- 14522226 TI - Comorbid symptom patterns in female juvenile offenders. PMID- 14522227 TI - Work and disproportionation for aqueous plutonium. AB - The relation of two plutonium work integrals has recently been illustrated. One of the integrals applies to the work of disproportionation of tetravalent plutonium in 1 M acid and the other to the work of oxidation of plutonium from the trivalent to a higher oxidation state. This paper generalizes the disproportionation work integral so that it can be applied to tetravalent plutonium at any acid concentration. An equation is provided that can be used to verify work estimations obtained by integration. It applies to oxidation and disproportionation processes and it is easy to use. PMID- 14522228 TI - Preparation of carrier-free 7Be by ion-exchange following charged particle and photonuclear reactions. AB - Carrier-free 7Be has been produced by 7Li(p,n)7Be and 10B(gamma,p2n)7Be (as well as 11B(gamma,p3n)7Be) reactions using a cyclotron and electron linear accelerator, respectively. Radiochemical methods for purification of the carrier free radioactive 7Be isotope from the irradiated lithium and boron compounds have been investigated. A simple separation scheme is proposed. PMID- 14522230 TI - A new technetium-99m labeled isonitrile complex 99mTc-TMCHI as a potential blood pool imaging agent. AB - A new isonitrile ligand 3,3,5-trimethyl cyclohexylisonitrile (TMCHI) and its copper salt [Cu(TMCHI)4BF4] were synthesized and characterized by IR and elemental analysis. 99mTc-TMCHI is prepared by direct labeling method with high RCP. In vitro protein binding in albumin, blood retention and biodistribution of 99mTc-TMCHI in mice indicated that it is mainly accumulated and maintained in blood with a high protein binding rate and low washout rate. The target/non target ratios in mice are excellent and suggest its use as a cardiac blood pool agent. PMID- 14522229 TI - Automated preparation of 188Re-labeled radiopharmaceuticals for endovascular radiation therapy. AB - We have developed an automated system for the preparation of highly concentrated 188Re-perrhenate, diethylenetriamine pentaacetic acid (DTPA) and mercaptoacetyltriglycine (MAG3). The three procedural steps include concentration of 188Re-perrhanerate, chelation and purification and sterilization. The steps are operated by a small micro-controller. The eluted 188Re-perrhenate of 15 GBq/18 ml from 37 GBq 188W/188Re-generator was concentrated to 1.2 ml in 10 +/- 0.5 min with a recovery yield of 95 +/- 1.5%. We obtained the highest radiochemical yield of 95.4 +/- 2.8% and 98.5 +/- 1.2% for 188Re-DTPA and MAG3 at the oil bath temperatures of 95-97 degrees C and 93-97 degrees C, respectively. PMID- 14522232 TI - A rapid method for alpha-spectrometric analysis of radium isotopes in natural waters using ion-selective membrane technology. AB - An alpha-spectrometric method for the rapid determination of radium isotopes (223Ra, 224Ra and 226Ra) in environmental samples is presented. Using Empore Radium Rad Disks complete separation of the target radionuclides is achieved. The high selectivity of these Rad Disks allows the straightforward use of 225Ra as yield tracer. Chemical yield is up to 92 +/- 9%. The chemical procedure can be accomplished within 5h. Alpha-sources show energy resolution in the range of typically 26-40 keV (FWHM). Despite minimal thickness of the sources no significant radon (Rn) losses can be observed. PMID- 14522231 TI - Aromatic L-amino acid decarboxylase (AAAD) inhibitors as carcinoid tumor-imaging agents: synthesis of 18F-labeled alpha-fluoromethyl-6-fluoro-m-tyrosine (FM-6 FmT). AB - The aromatic L-amino acid decarboxylase (AAAD) enzyme is significantly upregulated in neuroendocrine tumors and, thus, would be a good target for PET imaging agents. Alpha-fluoromethyl-DOPA (FMDOPA) is one of the most potent irreversible AAAD inhibitor and its non-catechol derivative, alpha-fluoromethyl-m tyrosine (FMmT), is a promising AAAD imaging agent. We synthesized FMmT and its direct electrophilic fluorination provided a mixture of products identified by NMR analysis after HPLC purification as 6-fluoro-, 2-fluoro- and 2,6-difluoro derivatives of FMmT. Using rat striatal homogenates, alpha-fluoromethyl-6-fluoro m-tyrosine (FM-6-FmT) was found to have AAAD inhibitory activity comparable to that of FMDOPA. Electrophilic radiofluorination of FMmT using [18F]AcOF gave 18F labeled 6-fluoro-, 2-fluoro- and 2,6-difluoro-FMmT derivatives in 22.0%, 21.9% and 8.5% radiochemical yields, respectively. Based on its proposed mechanism of inhibition, FM-6-[18F]FmT is expected to irreversibly bind to AAAD and, hence, could be used as a PET agent to image tumors of endocrine origin containing high concentrations of AAAD. Since FM-6-FmT lacks the catechol moiety, it is expected to be better than FMDOPA since it is not a substrate for catechol-O methyltransferase. PMID- 14522233 TI - Preconcentration of radium isotopes from natural waters using MnO2 Resin. AB - We have characterized "MnO2 Resin," a new resin developed by the PG Research Foundation, for radium adsorption over wide ranges of pH, reaction times and salt concentrations. We show that the sorption of 133Ba (used as a proxy for Ra) is highly dependent on pH with the most useful range from pH 4 to 8. The surface layers of the Mn oxides apparently become more positively charged under acidic conditions (below pH 4), which prevents diffusion of positively charged alkaline earth species (e.g. Ba2+, Ra2+) into the sorption sites. Adsorption at higher pH is thought to be inhibited because of carbonate complexation. We found that the sorption characteristics for radium onto MnO2 Resin are especially favorable for low-salinity waters but the sorption is still very satisfactory for highly salted solutions (KD=2.8x10(4) in both cases) but with slower kinetics. For analytical purposes, both column and pump experiments showed high recoveries with no measurable discrimination between Ra and Ba regardless of flow rates in fresh water. Seawater tests showed that recoveries of Ra and Ba are lower than fresh water at elevated flow rates with Ra adsorption higher than Ba at flow rates above 10 ml/min. PMID- 14522235 TI - Towards absolute activity measurements by ionisation chambers using the PENELOPE Monte-Carlo code. AB - The Monte-Carlo code PENELOPE for Ionisation Chamber Simulation Method has been applied for the calculation of ionisation-chamber (IC) calibration factors. Measuring only a few radionuclides well selected within the relevant energy range, and determining an adjustable parameter, commonly used radionuclides can be measured without any specific calibration. The simulation revealed a discontinuity in the IC response as a function of photon energy and its dependence on the chemical composition of the radioactive solution. PMID- 14522234 TI - Determination of neutron flux distribution in an Am-Be irradiator using the MCNP. AB - A neutron irradiator has been assembled at IPEN facilities to perform qualitative quantitative analysis of many materials using thermal and fast neutrons outside the nuclear reactor premises. To establish the prototype specifications, the neutron flux distribution and the absorbed dose rates were calculated using the MCNP computer code. These theoretical predictions then allow one to discuss the optimum irradiator design and its performance. PMID- 14522236 TI - Radon, thoron and their progenies measured in different dwelling rooms and reference atmospheres by using CR-39 and LR-115 SSNTD. AB - Detection efficiencies of the CR-39 and LR-115 type II solid state nuclear track detectors (SSNTD) for alpha-particles emitted by radon, thoron and their decay products inside the air of different dwelling rooms and in various reference atmospheres were determined by using a Monte Carlo computer code. Alpha- and beta activities per unit volume of air due to radon, thoron and their progenies were measured in the studied atmospheres by exploiting data obtained for the detection efficiencies of the SSNTD and measuring the resulting track density rates. Equilibrium factors between radon and its progeny and thoron and its daughters were evaluated in the studied atmospheres. PMID- 14522237 TI - Calibration of diffusion barrier charcoal detectors using a semi-empirical expression. AB - Several calibration settings of diffusion barrier charcoal canister (DBCC) detectors for measuring radon concentration in air were studied. A set of functions and graphs were developed for relations between radon concentration in air and adsorbed activity in DBCC, when calibrated in small chambers. Both the integration time for 10% of DBCC of a batch, and the radon adsorption coefficient for the activated charcoal used in these detectors, were determined. Thus, a semi empirical expression for detector calibration was applied. PMID- 14522238 TI - Radioelement distributions in the Proterozoic granites and associated pegmatites of Gabal El Fereyid area, Southeastern Desert, Egypt. AB - Lithologically, the rock types in the Gabal El Fereyid area are dominantly granites with minor amounts of pegmatites. The granites range in composition from tonalite to granite-adamallite with minor acidic dikes, quartz and pegmatite veins. The granite-adamallite is peraluminous and formed as a result of partial melting of amphibole-bearing rocks at depths of approximately 24-30 km and at temperatures of 800-950 degrees C. Among the different rock types, the muscovite rich pegmatites had the highest U and Th contents (66 and 38 ppm on average, respectively). The high level of radioactivity in pegmatites is attributed to the presence of the radioactive minerals thorianite, uranophane and allanite as confirmed by XRD analysis. Binary relations of Zr/U, Zr/Th, Ce/U and Ce/Th against either U or Th in the granite-adamellite exhibit significant negative correlations indicating that both elements are not preferentially hosted in the accessory minerals phases such as zircon and monazite, but could be associated with major forming minerals such as biotite, muscovite, plagioclase and quartz, or U is situated within labile sites within granite. The uranium and thorium enrichment in the pegmatites is a two-stage process. The primary stage is magmatic whereas the secondary enrichment is from hydrothermal concentration. The magmatic U and Th are indicated by the presence of thorianite and allanite, whereas evidence of hydrothermal mineralization is the alteration of rock-forming minerals such as feldspar and the formation of secondary minerals such as uranophane and pyrite. PMID- 14522239 TI - Role of ubiquitin-mediated proteolysis in the pathogenesis of neurodegenerative disorders. AB - Intraneuronal inclusions containing ubiquitylated filamentous protein aggregates are a common feature of many of the major human neurodegenerative disorders, including Alzheimer's and Parkinson's disease. Loss of function mutations in enzymes of the ubiquitin conjugation/deconjugation pathway are sufficient to cause familial forms of neurodegenerative diseases, suggesting that failure of ubiquitin-mediated proteolysis could also be central to inclusion formation in the more common sporadic cases. Examination of ubiquitin-positive inclusions at the protein level provides evidence of attempted proteasomal proteolysis, however close inspection of the temporal aspects of inclusion formation indicates that ubiquitylation is probably a late event. In this regard, the presence of ubiquitin within inclusions of idiopathic neurodegenerative disorders may indicate not a primary dysfunction of ubiquitin-mediated proteolysis, but rather a secondary, presumably protective cellular response. Within this model, other factors are likely to be initiating in inclusion biogenesis. Consistent with these proposals, non-ubiquitylated forms of the principal ubiquitylated components of Alzheimer's disease neurofibrillary tangles and Parkinson's disease Lewy bodies, tau and alpha-synuclein proteins, respectively, can be degraded by proteasomes in a pathway which does not have an absolute requirement for ubiquitylation. Inhibition of proteasome function in the pathological state, as has been reported in both Alzheimer's and Parkinson's disease, could therefore contribute both to accumulation of non-ubiquitylated forms of aggregation-prone neuronal proteins, as well as impaired clearance of ubiquitylated aggregates. PMID- 14522240 TI - Caspases, apoptosis and aging. AB - Caspases are a group of cysteine dependent aspartate-specific proteases. Originally found as the homolog of Ced-3 in C. elegans, 14 caspases have now been identified in mammals to date. Caspases play important roles in both the intrinsic and extrinsic apoptotic pathways and interact with the non-caspase apoptotic pathways. A number of recent published observations have suggested a strong association between apoptosis, age-related diseases and aging. Findings from our group and others reveal a strong correlation between alterations in caspase activity and aging. In this view point, we summarize current knowledge of the connection between caspases and aging observed in a variety of model systems from cultured cells in vitro to the in vivo models of rodents and humans. PMID- 14522241 TI - Neuronal and microglial cathepsins in aging and age-related diseases. AB - It has been long believed that cathepsins compensate for each other because of their overlapping substrate specificities. However, there is increasing evidence that disturbance of the normal balance of their enzymatic activities is the first insult in brain aging and age-related diseases. The imbalance of cathepsins may further cause age-related neuropathological changes such as accumulation of autophagic vacuoles and the formation of ceroid-lipofuscin leading to neuronal dysfunction and damage. Leakage of cathepsins due to the fragility of lysosomal membranes during aging also contributes to neurodegeneration. Furthermore, the deficiency of cathepsin D has been recently revealed to provoke a novel type of lysosomal storage disease associated with massive neurodegeneration. In these animals, microglia are activated to initiate inflammatory and cytotoxic responses by binding and phagocytosis of storage neurons. Activated microglia also release some members of cathepsins to induce neuronal death by degrading extracellular matrix proteins. Thus the microglial activation possibly through sensing neuronal storage may also be an important causative factor for neurodegeneration in lysosomal storage diseases and age-related diseases such as Alzheimer's disease. This review describes the pathological roles of neuronal and microglial cathepsins in brain aging and age-related diseases. PMID- 14522242 TI - Aging, lipofuscin formation, and free radical-mediated inhibition of cellular proteolytic systems. AB - Alterations in a wide array of physiological functions are a normal consequence of aging. Importantly, aged individuals exhibit an enhanced susceptibility to various degenerative diseases and appear less able than their young and adult counterparts to withstand (patho)physiological stress. Elucidation of mechanisms at play in the aging process would benefit the development of effective strategies for enhancing the quality of life for the elderly. It is likely that decrements in cellular and physiological function that occur during aging are the net result of numerous interacting factors. The current review focuses on the potential contribution(s) of free radical-mediated modifications to protein structure/function and alterations in the activities of two major proteolytic systems within cells, lysosomes and the proteasome, to the age-dependent accumulation of fluorescent intracellular granules, termed lipofuscin. Specifically, aging appears to influence the interplay between the occurrences of free radical-derived modifications to protein and the ability of cells to carry out critical proteolytic functions. We present immunochemical and ultrastructural evidence demonstrating the occurrence of a fluorescent protein cross-link derived from free radical-mediated reaction(s) within lipofuscin granules of rat cerebral cortex neurons. In addition, we provide evidence that a fluorophore-modified protein present in lipofuscin granules is the alpha subunit of F1F0-ATP synthase, a mitochondrial protein. It has previously been shown that protein(s) bearing this particular fluorescent cross-link are resistant to proteolysis and can inhibit the proteasome in a non-competitive fashion (J. Biol. Chem. 269 (1994a) 21639; FEBS Lett. 405 (1997) 21). Therefore, the current findings demonstrate that free radical-mediated modifications to protein(s) that lead to the production of inhibitor(s) of cellular proteolytic systems are present on specific protein components of lipofuscin. In addition, the mitochondrial origin of one of these proteins indicates specific intracellular pathways likely to be influenced by free radical events and participate in the formation of lipofuscin. The results of these studies are related to previous in vitro and in vivo observations in the field, thus shedding light on potential consequences to cellular function. In addition, future research directions suggested by the available evidence are discussed. PMID- 14522243 TI - The calpains in aging and aging-related diseases. AB - Calpains are a family of calcium-dependent cysteine proteases under complex cellular regulation. By making selective limited proteolytic cleavages, they modulate the activity of enzymes, including key signaling molecules, and induce specific cytoskeletal rearrangements, accounting for their roles in cell motility, signal transduction, vesicular trafficking and structural stabilization. Calpain activation has been implicated in various aging phenomena and diseases of late life, including cataract formation, erythrocyte senescence, diabetes mellitus type 2, hypertension, arthritis, and neurodegenerative disorders. The early and pervasive involvement of calpains in Alzheimer's disease potentially influences the development of beta-amyloid and tau disturbances and their consequences for neurodegeneration and neuronal cell loss. PMID- 14522244 TI - Immunoproteasomes and immunosenescence. AB - Aging is a complex process which is accompanied with the decline and the reshaping of different functions of the body. In particular the immune system is characterized, during ageing (immunosenescence) by a remodeling of innate immunity (well preserved, up-regulated) and clonotypical immunity (severely altered) and by the occurrence of a chronic inflammatory process (inflammaging) which are, at least in part, genetically controlled. In this scenario, it can be anticipated that a crucial role is played by age-related structural and functional alterations and modifications of proteasomes and immunoproteasomes, the last being a key component of antigen processing and MHC class I antigen presentation. A variety of experimental data are available, suggesting that proteasomes are affected by age, and that in centenarians they are relatively preserved. On the contrary, few data are available on immunoproteasomes, likely as a consequence of the poverty of suitable cellular models. Lymphoblastoid cell lines from EBV immortalized B cells from old donors is envisaged as a possible model for the study of immunoproteasomes in humans and their changes with age. Thus, basic questions such as those related to possible consequences, for immune responses in infectious diseases and cancer, of age-related alterations of antigen processing and presenting, change with age of self-antigen repertoire, and the genetic basis of immunoproteasome activity and its change with age, remain largely unanswered. PMID- 14522245 TI - Monitoring the ubiquitin/proteasome system in conformational diseases. AB - Controlled proteolysis of regulatory or aberrant proteins by the ubiquitin/proteasome system is indispensable for cell viability. Conformational diseases such as Alzheimer's, Parkinson's and Huntington's disease are characterised by the accumulation of misfolded or aggregation-prone proteins. Since these proteins are typical substrates of the ubiquitin/proteasome system, it is not surprising that various models propose impairment of this system as a contributing factor to the pathology of conformational disorders. The complex nature of the ubiquitin/proteasome system and its universal role in cell physiology however turns evaluation of these attractive hypotheses into a major challenge. Several reporter substrates for the ubiquitin/proteasome system have recently been developed to facilitate functional studies of the system in living cells. In this review, we will discuss these new tools as well as the proteins associated with conformational disease that have been studied with these reporters. PMID- 14522246 TI - Calcium-dependent and aspartyl proteases in neurodegeneration and ageing in C. elegans. AB - Proteolytic mechanisms have been implicated in the process of ageing, and in many neurodegenerative disorders such as Alzheimer's, Parkinson's and Huntington's diseases, which are most prevalent in old age. Simple model organisms such as the nematode Caenorhabditis elegans and the fruit fly Drosophila melanogaster, which offer the prowess of sophisticated genetic approaches, have contributed to our understanding of ageing and neurodegeneration. Intensive research in these systems has resulted in detailed models of the ageing process, and also of several neurodegenerative diseases, which recapitulate same aspects of the human pathologies. Inappropriate cell death is a major component of these and other devastating conditions such as stroke. The dissection of the molecular mechanisms underlying the process of cell degeneration in ageing is of utmost importance. Evidence from investigations in C. elegans implicates deregulated proteolysis as one major determinant of cellular destruction in neurodegeneration and ageing, and suggests that the process depends critically on the activation of calcium dependent, calpain proteases and lysosomal aspartyl proteases. Apart from shedding light on important but inadequately understood facets of such phenomena, these discoveries hold promise for developing novel, effective intervention strategies aiming to ameliorate or even counter inappropriate cell death. PMID- 14522247 TI - Environmental tobacco smoke, carcinogenesis, and angiogenesis: a double whammy? AB - In this issue of Cancer Cell, Zhu and coworkers demonstrate that environmental tobacco smoke (ETS) results in tumor angiogenesis, and provide evidence that the responsible factor in ETS is nicotine. While a weak carcinogen, nicotine promotes carcinogenesis by a number of different mechanisms. The authors' findings provide further evidence of the harmful effects of ETS and indicate the desirability of reducing exposure to it. PMID- 14522248 TI - Cell cycle progression without cyclin E/CDK2: breaking down the walls of dogma. AB - G1 is the phase of the cell cycle wherein the cell is responsive to growth factor dependent signals. As such, G1 regulation is frequently disrupted in cancer through deregulation of cyclin/CDK activity; deregulation of G1 phase provides tumorigenic cells with a growth advantage. Cyclin E, the regulatory cyclin for CDK2, is considered a requisite regulator of G1 progression. Cyclin E is overexpressed in cancer, suggesting that cyclin E/CDK2 deregulation contributes to tumorigenesis. Two papers now challenge both the concept that cyclin E/CDK2 is a requisite component of the cell cycle machine and efforts to develop cyclin E/CDK2 inhibitors as antiproliferative therapeutics. PMID- 14522249 TI - A big step in the study of small cell lung cancer. AB - A rationally designed, conditional p53 and Rb allele-based and lung-targeted mouse model of human small cell lung cancer (SCLC) provides the cancer research community with a valid and important new tool to use in translational research against this deadly disease. PMID- 14522250 TI - Cell signaling and cancer. AB - During the course of tumor progression, cancer cells acquire a number of characteristic alterations. These include the capacities to proliferate independently of exogenous growth-promoting or growth-inhibitory signals, to invade surrounding tissues and metastasize to distant sites, to elicit an angiogenic response, and to evade mechanisms that limit cell proliferation, such as apoptosis and replicative senescence. These properties reflect alterations in the cellular signaling pathways that in normal cells control cell proliferation, motility, and survival. Many of the proteins currently under investigation as possible targets for cancer therapy are signaling proteins that are components of these pathways. The nature of these signaling pathways and their roles in tumorigenesis were the subject of a recent Beatson International Cancer Conference. PMID- 14522251 TI - Prolyl isomerase Pin1 as a molecular target for cancer diagnostics and therapeutics. PMID- 14522252 TI - Induction of small cell lung cancer by somatic inactivation of both Trp53 and Rb1 in a conditional mouse model. AB - Small cell lung cancer (SCLC) is a highly aggressive human tumor with a more than 95% mortality rate. Its ontogeny and molecular pathogenesis remains poorly understood. We established a mouse model for neuroendocrine (NE) lung tumors by conditional inactivation of Rb1 and Trp53 in mouse lung epithelial cells. Mice carrying conditional alleles for both Rb1 and Trp53 developed with high incidence aggressive lung tumors with striking morphologic and immunophenotypic similarities to SCLC. Most of these tumors, which we designate MSCLC (murine small cell lung carcinoma), diffusely spread through the lung and gave rise to extrapulmonary metastases. In our model, inactivation of both Rb1 and p53 was a prerequisite for the pathogenesis of SCLC. PMID- 14522253 TI - Second hand smoke stimulates tumor angiogenesis and growth. AB - Exposure to second hand smoke (SHS) is believed to cause lung cancer. Pathological angiogenesis is a requisite for tumor growth. Lewis lung cancer cells were injected subcutaneously into mice, which were then exposed to sidestream smoke (SHS) or clean room air and administered vehicle, cerivastatin, or mecamylamine. SHS significantly increased tumor size, weight, capillary density, VEGF and MCP-1 levels, and circulating endothelial progenitor cells (EPC). Cerivastatin (an inhibitor of HMG-coA reductase) or mecamylamine (an inhibitor of nicotinic acetylcholine receptors) suppressed the effect of SHS to increase tumor size and capillary density. Cerivastatin reduced MCP-1 levels, whereas mecamylamine reduced VEGF levels and EPC. These studies reveal that SHS promotes tumor angiogenesis and growth. These effects of SHS are associated with increases in plasma VEGF and MCP-1 levels, and EPC, mediated in part by isoprenylation and nicotinic acetylcholine receptors. PMID- 14522254 TI - Dimerization of MLL fusion proteins immortalizes hematopoietic cells. AB - MLL fusion proteins are leukemogenic, but their mechanism is unclear. Induced dimerization of a truncated MLL immortalizes bone marrow and imposes a reversible block on myeloid differentiation associated with upregulation of Hox a7, a9, and Meis1. Both dimerized MLL and exon-duplicated MLL are potent transcriptional activators, suggesting a link between dimerization and partial tandem duplication of DNA binding domains of MLL. Dimerized MLL binds with higher affinity than undimerized MLL to a CpG island within the Hox a9 locus. However, MLL-AF9 is not dimerized in vivo. The data support a model in which either MLL dimerization/exon duplication or fusion to a transcriptional activator results in Hox gene upregulation and ultimately transformation. PMID- 14522255 TI - Prostate-specific deletion of the murine Pten tumor suppressor gene leads to metastatic prostate cancer. AB - The murine Pten prostate cancer model described in this study recapitulates the disease progression seen in humans: initiation of prostate cancer with prostatic intraepithelial neoplasia (PIN), followed by progression to invasive adenocarcinoma, and subsequent metastasis with defined kinetics. Furthermore, while Pten null prostate cancers regress after androgen ablation, they are capable of proliferating in the absence of androgen. Global assessment of molecular changes caused by homozygous Pten deletion identified key genes known to be relevant to human prostate cancer, including those "signature" genes associated with human cancer metastasis. This murine prostate cancer model provides a unique tool for both exploring the molecular mechanism underlying prostate cancer and for development of new targeted therapies. PMID- 14522257 TI - No more business as usual in China. PMID- 14522256 TI - Myc-driven murine prostate cancer shares molecular features with human prostate tumors. AB - Increased Myc gene copy number is observed in human prostate cancer. To define Myc's functional role, we generated transgenic mice expressing human c-Myc in the mouse prostate. All mice developed murine prostatic intraepithelial neoplasia followed by invasive adenocarcinoma. Microarray-based expression profiling identified a Myc prostate cancer expression signature, which included the putative human tumor suppressor NXK3.1. Human prostate tumor databases revealed modules of human genes that varied in concert with the Myc prostate cancer signature. This module includes the Pim-1 kinase, a gene known to cooperate with Myc in tumorigenesis, and defines a subset of human, "Myc-like" human cancers. This approach illustrates how genomic technologies can be applied to mouse cancer models to guide evaluation of human tumor databases. PMID- 14522258 TI - ACT now--with caution--for malaria treatments. PMID- 14522259 TI - Is there a role for nitric oxide in hyperlactataemia syndromes and mitochondrial dysfunction associated with HIV therapy? PMID- 14522260 TI - Polyomavirus BK. AB - Polyomavirus hominis 1, better known as BK virus (BKV), infects up to 90% of the general population. However, significant clinical manifestations are rare and limited to individuals with impaired immune functions. BKV has been associated with diverse entities such as haemorrhagic cystitis, ureteric stenosis, vasculopathy, pneumonitis, encephalitis, retinitis, and even multi-organ failure. In addition, BKV has been implicated in autoimmune disease and possibly cancer. Due to high prevalence and frequent reactivation, the role of BKV in some of these pathologies has been difficult to define. Development of BKV diseases is likely to require complementing determinants in the host, the target organ, and possibly the virus, that are subject to modulators such as immunosuppression. These complex aspects are highlighted in polyomavirus-associated nephropathy (PAN), an emerging disease in renal allograft recipients that may jeopardise the progress in renal transplantation accomplished in the past 10 years. Intervention is difficult due to the lack of specific antivirals and relies mostly on improving immune control. Diagnostic strategies using urine cytology and BKV load measurements in plasma have led to earlier diagnosis of PAN, which increased the success rate of intervention. Case series suggest that cidofovir might be effective, especially when combined with reduced immunosuppression. PMID- 14522261 TI - Diagnosis and treatment of tuberculosis in children. AB - There has been a recent global resurgence of tuberculosis in both resource limited and some resource-rich countries. Several factors have contributed to this resurgence, including HIV infection, overcrowding, and immigration. Childhood tuberculosis represents a sentinel event in the community suggesting recent transmission from an infectious adult. The diagnosis of tuberculosis in children is traditionally based on chest radiography, tuberculin skin testing, and mycobacterial staining/culture although these investigations may not always be positive in children with tuberculosis. Newer diagnostic methods, such as PCR and immune-based methods, are increasingly being used although they are not widely available and have a limited role in routine clinical practice. Diagnostic approaches have been developed for use in resource-limited settings; however, these diagnostic methods have not been standardised and few have been validated. Short-course, multidrug treatment has been adopted as standard therapy for adults and children with tuberculosis, with or without directly observed therapy. Compliance is a major determinant of the success of drug treatment. Although uncommon in children, multidrug-resistant tuberculosis is also increasing and treatment will often involve longer courses of therapy with second-line antituberculosis drugs. Treatment of latent infection and chemoprophylaxis of young household contacts is also recommended for tuberculosis prevention, although this may not always be carried out, particularly in high incidence areas. PMID- 14522262 TI - Diagnostic accuracy of nucleic acid amplification tests for tuberculous meningitis: a systematic review and meta-analysis. AB - Conventional tests are not always helpful in making a diagnosis of tuberculous meningitis. We did a systematic review and meta-analysis to establish the summary accuracy of nucleic acid amplification (NAA) tests for tuberculous meningitis. We searched six electronic databases and contacted authors, experts, and manufacturers. Measures of diagnostic accuracy were pooled using a random effects model. 49 studies met our inclusion criteria. The summary estimates in 14 studies with commercial NAA tests were: sensitivity 0.56 (95% CI 0.46, 0.66), specificity 0.98 (0.97, 0.99), positive likelihood ratio 35.1 (19.0, 64.6), negative likelihood ratio 0.44 (0.33, 0.60), and diagnostic odds ratio 96.4 (42.8, 217.3). In the 35 studies with in-house ("home-brew") tests, the summary accuracy could not be established with confidence because of wide variability in test accuracy. On current evidence, commercial NAA tests show a potential role in confirming tuberculous meningitis diagnosis, although their overall low sensitivity precludes the use of these tests to rule out tuberculous meningitis with certainty. PMID- 14522263 TI - Tryptophan depletion and HIV infection: a metabolic link to pathogenesis. AB - HIV-1-infected patients have low circulating tryptophan concentrations despite evidence of adequate dietary intake of this essential amino acid. A chronic increase in inducible tryptophan oxidation is the basis of HIV-1-associated tryptophan depletion. This metabolic process results in the irretrievable loss of tryptophan molecules from the available pool. Such sustained disruption of normal tryptophan metabolism over time disturbs the many metabolic processes involving this amino acid, and has been implicated in some features of AIDS pathogenesis. Normal T-cell function is adversely affected by tryptophan depletion, but the extent of the effect in HIV-1-infected patients is still unclear. Attempting to directly supplement tryptophan is not advised given the potential increase in circulating concentrations of neurotoxic intermediates. Although only preliminary data are available, evidence suggests that antiretroviral and nicotinamide treatments can boost plasma tryptophan concentrations in HIV-1-infected patients and impact the secondary effects of tryptophan depletion. Additional study of this metabolism could lead to improved treatment strategies for patients with HIV infection. In this review I focus on the potential links between disturbed tryptophan metabolism and pathogenesis. PMID- 14522266 TI - Following in Soper's footsteps: northeast Brazil 63 years after eradication of Anopheles gambiae. AB - Sub-Saharan Africa has long suffered under the yoke of the Anopheles gambiae mosquito, but for northeast Brazil (figure 1) its arrival over 60 years ago was a new and horrifying experience. This African mosquito is an exceptionally effective malaria vector because it is well adapted to feeding upon people and to exploiting aquatic habitats associated with our daily activities. Anopheles gambiae sensu lato probably accounts for most of the world's malaria deaths and socioeconomic burden. Fortunately, the Brazilian experience had a happy ending. The prospect of A gambiae spreading across much of the Americas motivated a ruthlessly effective response that deserves a special and heroic place in the annals of public health. Building on the successes and infrastructure of the Yellow Fever Service for Aedes aegypti elimination, the Rockefeller Foundation and Brazilian government collaborated to form a new Malaria Service of the Northeast. This new entity rolled the invader back into oblivion with an aggressive eradication campaign, focusing primarily upon larviciding of all potential habitats. The driving force of this endeavour was an enigmatic man called Fred Soper whose sheer will and determination was a key element in this success, and a source of inspiration today (see Killeen GF, et al. Eradication of Anopheles gambiae from Brazil: lessons for malaria control in Africa? Lancet Infect Dis 2002; 2: 618-27). I recently took an opportunity to fulfil a long-held dream and follow in some of Soper's footsteps. Tired of gazing at yellowing maps like figure 1, I went to see the northeast of Brazil for myself. PMID- 14522267 TI - W Michael Scheld--President of the Infectious Diseases Society of America interview by Pam Das. PMID- 14522264 TI - Hantavirus infections in Europe. AB - Hantaviruses are enveloped RNA viruses each carried by a specific rodent species. Three hantaviruses, Puumala, Dobrava, and Saaremaa viruses, are known to cause haemorrhagic fever with renal syndrome. In Europe. Puumala causes a generally mild disease, nephropathia epidemica, which presents most commonly with fever, headache, gastrointestinal symptoms, impaired renal function, and blurred vision, whereas Dobrava infections often also have haemorrhagic complications. There are few available data about the clinical picture of confirmed Saaremaa infections, but epidemiological evidence suggests that it is less pathogenic than Dobrava, and that Saaremaa infections are more similar to nephropathia epidemica caused by Puumala. Along with its rodent host, the bank vole (Clethrionomys glareolus), Puumala is reported throughout most of Europe (excluding the Mediterranean region), whereas Dobrava, carried by the yellow-necked mouse (Apodemus flavicollis), and Saaremaa, carried by the striped field mouse (Apodemus agrarius), are reported mainly in eastern and central Europe. The diagnosis of acute hantavirus infection is based on the detection of virus-specific IgM. Whereas Puumala is distinct, Dobrava and Saaremaa are genetically and antigenically very closely related and were previously thought to be variants of the same virus. Typing of a specific hantavirus infection requires neutralisation antibody assays or reverse transcriptase PCR and sequencing. PMID- 14522270 TI - Going off tomatoes. PMID- 14522271 TI - The value of surveys for cataract and refractive surgery. PMID- 14522272 TI - U.s. Code of Federal Regulations. PMID- 14522273 TI - Terms for photic phenomena. PMID- 14522274 TI - Contrast sensitivity and spatial-frequency spectrum after refractive surgery. PMID- 14522275 TI - Pseudophakic retinal detachments. PMID- 14522276 TI - Anterior capsule contraction. PMID- 14522277 TI - Consultation section. Refractive surgical problem. PMID- 14522282 TI - Surgical technique for suture fixation of an acrylic intraocular lens in the absence of capsule support. AB - We present a technique for suture fixation of an intraocular lens (IOL) in the absence of capsule support. This technique is useful for the treatment of aphakia and the management of IOL complications or cataract surgery when all capsule support is lost. PMID- 14522283 TI - Simplified small-incision peripheral iris fixation of an AcrySof intraocular lens in the absence of capsule support. AB - In the absence of capsule and zonule support, a modified insertion technique allows simple peripheral iris fixation of a posterior chamber 3-piece acrylic intraocular lens (IOL) via a small incision for secondary implantation. A bimanual unfolding maneuver creates pupillary capture of the optic to temporarily stabilize the IOL. Peripheral modified McCannel sutures fixate the flexible monofilament poly(methyl methacrylate) haptics to the posterior peripheral iris surface, after which the optic is safely prolapsed into the posterior chamber. PMID- 14522284 TI - Modified capsular tension ring for patients with congenital loss of zonular support. AB - PURPOSE: To evaluate the results of implantation of a modified capsular tension ring (MCTR) and a posterior chamber intraocular lens (PC IOL) in patients with congenitally subluxated crystalline lenses. SETTING: Cincinnati Eye Institute, Cincinnati, Ohio, USA. METHODS: Ninety eyes of 57 patients with congenital loss of zonular support (Weill-Marchesani syndrome, idiopathic ectopia lentis, and Marfan's syndrome) had phacoemulsification with PC IOL and MCTR implantation. The preoperative examination included best corrected visual acuity (BCVA) and the presence or absence of phacodonesis, lens decentration, and vitreous prolapse. The postoperative evaluation included BCVA and the presence or absence of pseudophacodonesis, PC IOL centration, and posterior capsule opacification (PCO). RESULTS: At the last postoperative examination, the BCVA was 20/40 or better in 80 eyes (88.9%); 1 eye (1.1%) lost 1 line of acuity. Preoperatively, 18 eyes (20%) had phacodonesis; 1 eye had postoperative pseudophacodonesis. Decentration before surgery was present in 86 eyes (95.6%); 6 eyes (6.7%) developed late symptomatic PC IOL decentration a median of 17.84 months +/- 10.73 (SD) after surgery. Other complications were increased intraocular pressure (2.2%), persistent iritis (3.3%), broken suture (10.0%), retinal detachment (1.1%), and PCO (20.0%). CONCLUSIONS: Use of the MCTR resulted in centration of the capsular bag and PC IOL in 90 eyes with congenitally subluxated crystalline lenses. Fixation of a 9-0 polypropylene suture is recommended to decrease the risk for late suture breakage. PMID- 14522285 TI - Estimating residual stromal thickness before and after laser in situ keratomileusis. AB - PURPOSE: To determine the factor(s) that influences measurement of residual stromal thickness (RST) after laser in situ keratomileusis (LASIK) surgery. SETTING: Clinical office-based excimer laser refractive surgery center. METHODS: In this retrospective comparative interventional case study of 6235 eyes, ultrasonic corneal pachymetry was performed immediately before and after flap creation and immediately after laser ablation in the primary procedure and after 647 enhancements. Differences in the methods for calculating RST were compared statistically. RESULTS: Using the RST measured at enhancement as the actual RST, measurements of RST immediately after laser ablation underestimated residual thickness due to laser-induced stromal dehydration and microkeratome effects (P<.001). Estimates of RST using a "standard" or estimated flap thickness were less accurate predictors of residual thickness (P<.001) than use of the theoretical laser resection with a measured flap thickness (RST-4) (P =.78) or a modified flap thickness subtracted from the postoperative corneal thickness (RST 8) (P =.98), which provided the best RST estimates. CONCLUSIONS: Before LASIK, the best means of estimating RST is to subtract the theoretical laser resection obtained from the laser computer and the expected flap thickness normally obtained with a given microkeratome system from the preoperative central corneal thickness. After LASIK, the most accurate means of calculating RST is to subtract the original flap thickness from the postoperative central corneal thickness. PMID- 14522286 TI - Retinal image contrast and functional visual performance with aspheric, silicone, and acrylic intraocular lenses. Prospective evaluation. AB - PURPOSE: To compare the effects of an aspheric intraocular lens (IOL) (Tecnis Z 9000, Pharmacia Corp.) with those of conventional silicone (AA4207VF, Staar Surgical) and acrylic (AcrySof SA60AT, Alcon Surgical) lenses on retinal imaging and functional visual performance. SETTING: Eye Laser Center, Tucson, Arizona, USA. METHODS: In this prospective study, 221 eyes of 156 patients were randomly assigned to receive 1 of 3 IOLs. Visual acuity was measured preoperatively and 1 day, 1 and 3 weeks, and 1, 3, and 6 months postoperatively. Fundus photography and photopic and mesopic functional acuity contrast testing were performed preoperatively and 3 months postoperatively. RESULTS: The differences in the preoperative and postoperative spherical and astigmatic refractive errors and best corrected visual acuity among groups were not statistically significant. The postoperative uncorrected visual acuity was best in the aspheric group in the first month. Functional acuity contrast testing showed the aspheric IOL group had a 38% to 47% increase in photopic vision, 38% in photopic with glare, 43% to 100% in mesopic, and 9% to 100% in mesopic with glare. There was no increase in photopic vision in the acrylic group but increases of 38% in photopic with glare, 25% to 50% in mesopic, and 36% to 50% in mesopic with glare. The spherical silicone IOL group had no increase in contrast testing from preoperatively. Digital analysis of retinal imaging showed increased threshold luminance levels in the aspheric group (range 116 to 208 cd/m2) and a 4-fold increase in image contrast. CONCLUSIONS: All 3 IOLs led to improved visual acuity after cataract surgery. The aspheric IOL provided a significant improvement in retinal image contrast and visual performance measured by visual acuity and functional acuity contrast testing. This improvement was greatest in night vision and night vision with glare compared to the performance of conventional spherical silicone and acrylic IOLs. PMID- 14522287 TI - Comparison of myopes and hyperopes after laser in situ keratomileusis monovision. AB - PURPOSE: To compare the results of laser in situ keratomileusis (LASIK) monovision in myopes and hyperopes. SETTING: Private practice, Little Silver, New Jersey, USA. METHODS: In this retrospective study, 391 consecutive patients older than 40 years who had LASIK between December 1999 and June 2001 were examined. All patients were asked to complete a questionnaire, and study results are based on the analysis of data from patient charts correlated with questionnaire responses. RESULTS: Visual results were excellent in both groups, with all monovision patients achieving 20/30 acuity or better in the distance eye and J2 or better acuity in the near eye. Satisfaction scores were high in all groups. Statistical analysis indicated that visual results for distance were better in myopes than in hyperopes (P =.043), enhancements were higher in monovision (P =.04) and in hyperopia (P =.08), and satisfaction was higher among myopes (P =.012) and full distance patients (P =.002). Monovision hyperopes had the most difficulty with 6 side effects (P =.00). CONCLUSION: Hyperopic monovision was a viable but more problematic solution to correcting presbyopia than myopic monovision. PMID- 14522288 TI - Anterior corneal optical aberrations induced by laser in situ keratomileusis for hyperopia. AB - PURPOSE: To evaluate the effects of hyperopic laser in situ keratomileusis (LASIK) on anterior corneal higher-order aberrations (HOAs) (3rd to 6th orders). SETTING: Cullen Eye Institute, Baylor College of Medicine, Houston, Texas, USA. METHODS: In a retrospective study, 3rd-order to 6th-order HOAs from preoperative and 3-month postoperative corneal topographic maps of eyes that had hyperopic LASIK using the Visx Star S2 and Visx Star S3 laser systems were calculated. There were 20 eyes of 14 patients in each group; the optical and transition zones were 6.0 mm and 9.0 mm, respectively. The surgically induced changes in anterior corneal HOAs and the differences between the HOA changes in corneas treated with and without active eye tracking were analyzed. RESULTS: In both groups, hyperopic LASIK induced a significant reduction in 4th-order spherical aberration (SA) coefficients (Z4(0)) in the 3.0 mm and the 6.0 mm central zones, and the decrease was significantly correlated with increasing hyperopic correction (except in the Visx S3 group's central 3.0 mm zone). In the 6.0 mm zone in both groups, the root mean-square (RMS) of the SA (Z4(0) and Z6(0)) decreased significantly, whereas there were no significant changes in the total HOAs. By excluding Z4(0), the mean RMS values for the remaining 21 Zernike terms increased by more than 30% (both P<.016). A simulated increase in pupil size from 3.0 to 6.0 mm caused a significantly smaller increase in SA in postoperative corneas compared to the preoperative values. Differences between the S2 and S3 groups were minimal. CONCLUSIONS: Hyperopic LASIK decreased SAs and reduced the magnitude of the SA increase that occurred with pupil dilation. Higher-order aberrations excluding the 4th-order SA increased significantly. PMID- 14522289 TI - Fourier analysis of induced irregular astigmatism. Photorefractive keratectomy versus laser in situ keratomileusis in a bilateral cohort of hyperopic patients. AB - PURPOSE: To analyze corneal topographic data by Fourier analysis to determine differences in irregular astigmatism following spherical hyperopic correction by photorefractive keratectomy (PRK) or laser in situ keratomileusis (LASIK). SETTING: Department of Ophthalmology, St. Thomas' Hospital, London, United Kingdom. METHODS: Thirty-six eyes of 18 patients with moderate hyperopia had LASIK in 1 eye and PRK in the other eye. The flap was cut on a nasal hinge with a Moria LSK One microkeratome. The laser was a Summit SVS Apex Plus with an optical zone of 6.5 mm and a blending zone of 1.5 mm. Corneal topographic data were acquired with a TMS-1 topographer (Computed Anatomy Inc.) preoperatively and 1, 3, 6, and 12 months postoperatively. The ASCII files containing the dioptric power values were extracted and analyzed with custom-written software to extract the Fourier harmonics. RESULTS: The irregular astigmatism increased in both groups postoperatively, peaking at 3 months and then decreasing over the next 9 months. There was no statistically significant difference between the 2 groups at any time point (P<.05). The change in the topographically derived equivalent sphere showed undercorrection in both groups at all time points. Regular astigmatism showed a marginal statistically significant increase in the LASIK group at 12 months (P =.049). CONCLUSION: Irregular astigmatism, equivalent sphere, and regular astigmatism were not significantly different in the PRK and LASIK groups during the follow-up. Based on the corneal topography, the 2 procedures induced an equal amount of irregular astigmatism. PMID- 14522290 TI - Stromal haze after laser in situ keratomileusis: clinical and confocal microscopy findings. AB - PURPOSE: To report clinical and confocal microscopy characteristics of haze-like opacities in corneas after laser in situ keratomileusis (LASIK). SETTING: Department of Ophthalmology, Johann Wolfgang Goethe-University, Frankfurt am Main, Germany. METHODS: Eighteen eyes of 11 patients with clinically apparent corneal clouding were examined by slitlamp and confocal microscopy (Confoscan P4, Tomey) 1 to 9 months after primary LASIK or LASIK retreatment. RESULTS: Postoperative slitlamp examination showed faint, white, snowflake-like clouding at the interface level in all patients. One patient had folds and rather diffuse haze-like opacities. Confocal microscopy revealed highly reflective structures in the flap stroma and at the interface level in all patients, probably due to numerous activated keratocytes and their processes. The confocal microscopy appearance was similar to that of photorefractive keratectomy haze. CONCLUSION: Focal wound-healing reactions in the central flap stroma and interface resulting in significant keratocyte activation could be observed after LASIK. PMID- 14522291 TI - Residual cleaner after normal cleaning of laser in situ keratomileusis instruments. AB - PURPOSE: To determine whether residual cleaner could be detected in the rinse solution of surgical instruments after a standard cleaning protocol. SETTING: Magill Research Center for Vision Correction, Storm Eye Institute, Medical University of South Carolina, Charleston, South Carolina, USA. METHODS: The wavelength for maximum absorbance of 5 cleaners (Endozime) [The Ruhof Corp.], Enzol/Cidezyme [Advanced Sterilization Products], and Klenzyme [Steris Co.] enzymatic detergents; Palmolive Ultra dishwashing liquid [Colgate-Palmolive Co.]; and Universal concentrated surgical instrument cleaner and lubricant [B. Graczyk, Inc.]) was determined. Identically designed stainless-steel and titanium instruments were cleaned using a standardized protocol. Water temperatures of 25 degrees C (room temperature) or 40 degrees C (warm temperature) were used to rinse the instruments. The amount of residual cleaner in each rinse solution and remaining on each instrument at the completion of the cleaning procedure as a percentage of the total cleaner was determined. RESULTS: Residues of all cleaners were detected using a standard protocol involving rinse solutions at 25 degrees C. Increasing the temperature of the rinse solutions significantly reduced the cleaner residues (P<.05, Friedman repeated-measures analysis of variance on ranks test and Student-Newman-Keuls test) regardless of the instrument material. No significant difference was detected in the residual cleaners on stainless-steel and titanium instruments. CONCLUSIONS: Lower levels of cleaner residue were found on surgical instruments after a standard cleaning protocol using warm rinse water. Because cleaner residue has been reported to cause inflammation (eg, diffuse lamellar keratitis) after laser in situ keratomileusis (LASIK), it is advisable to use cleaners and cleaning protocols that result in acceptable cleaning without detectable levels of cleaner residue to avoid potentially harmful effects to the cornea after LASIK. PMID- 14522292 TI - Anterior and posterior capsulorhexis in pediatric cataract surgery with or without trypan blue dye: randomized prospective clinical study. AB - PURPOSE: To evaluate the clinical efficacy of trypan blue 0.1% dye (Blurhex) in creating a complete anterior and posterior capsulorhexis during pediatric cataract surgery. SETTING: Tertiary eyecare center, Postgraduate Institute of Medical Education and Research, Chandigarh, India. METHODS: In a prospective randomized study, an anterior (ACCC) and posterior (PCCC) continuous curvilinear capsulorhexis were performed in pediatric patients. In Group 1 (n = 19), the ACCC and PCCC were created without use of trypan blue dye. In Group 2 (n = 23), intracameral trypan blue dye was used to stain the anterior and posterior capsules. RESULTS: In Group 1, 14 eyes (73.6%) had a complete ACCC and 10 (52.6%) had a complete PCCC. In Group 2, 21 eyes (91.3%) had a complete ACCC and 19 (82.6%) had a complete PCCC. CONCLUSION: In cases of pediatric cataract, staining the anterior and posterior capsules with trypan blue 0.1% allowed recognition of capsule flaps and facilitated the creation of complete ACCCs and PCCCs. PMID- 14522293 TI - Single-piece acrylic intraocular lens implantation in children. AB - PURPOSE: To assess the short-term outcomes of single-piece acrylic intraocular lens (IOL) implantation in children by determining the incidence of postoperative visual axis opacification and the need for a second procedure to clear the axis, cell deposits on the IOL optic, posterior synechias, and IOL decentration. SETTING: Miles Center for Pediatric Ophthalmology, Storm Eye Institute, Medical University of South Carolina, Charleston, South Carolina, USA. METHODS: This retrospective case review comprised 43 consecutive implantations (33 patients) of a single-piece hydrophobic acrylic IOL (AcrySof SA30AL or SA60AT, Alcon). An analysis of 42 eyes with posterior capsulectomy and vitrectomy was performed. Eyes with traumatic cataract and secondary IOLs were excluded. RESULTS: Single piece acrylic IOLs were implanted in 42 eyes. The mean age was 33.5 months +/- 28.9 (SD) (range 0.5 to 110 months) and the mean follow-up, 12.0 +/- 8.2 months (range 1.0 to 27.5 months). Postoperative opacification of the visual axis occurred in 7 eyes (16.7%). Secondary surgical procedures were required in 5 eyes (11.9%). Lens deposits were observed in 8 eyes (19.0%) and synechias, in 5 eyes (11.9%). All IOLs were well centered postoperatively. CONCLUSION: The short-term data suggest implantation of the AcrySof single-piece hydrophobic acrylic IOL is safe in the pediatric eye. PMID- 14522294 TI - Successful implementation of a protocol for photorefractive keratectomy in children requiring anesthesia. AB - PURPOSE: To describe a protocol for treating children with photorefractive keratectomy (PRK) under general anesthesia and to review intraoperative and postoperative complications. SETTING: Institutional academic practice. METHODS: Nine patients between 3 years and 9 years of age were treated with PRK under general anesthesia for anisometropia with unilateral high myopia or high hyperopia and amblyopia of the affected eye. Induction of anesthesia and the surgical procedure were carried out in separate rooms. The laser beam was centered on the entrance pupil, and eye position was monitored throughout the procedure. Specific precautions were taken before and during the procedure to prevent unwanted effects of inhalational anesthetic agents on laser performance. RESULTS: All children did well, with no anesthesia-related or treatment-related complications. CONCLUSIONS: Our protocol for PRK under general anesthesia was effective and efficient in children who were unable to cooperate for the procedure using local anesthesia. It can be adapted for laser in situ keratomileusis and other refractive surgical procedures in children and uncooperative adults. PMID- 14522296 TI - Evaluation of quality of life in patients with cataract in Hong Kong. AB - PURPOSE: To evaluate the quality of life in patients in a public hospital in Hong Kong before and after cataract surgery using a new questionnaire. SETTING: Department of Ophthalmology, Tung Wah Eastern Hospital, Hong Kong, China. METHODS: One hundred ten Chinese patients having cataract extraction were evaluated for surgical outcomes and quality of life using a questionnaire modified to reflect the local culture and environment in Hong Kong. The questionnaire consisted of 20 questions divided into 4 domains: distance vision, near vision, social function, and cataract-related symptoms. The test-retest reliability of the questionnaire was assessed in another 15 patients by weighted kappa and internal consistency tested with the Cronbach alpha. Construct validity was tested by correlating visual acuity with quality of life and subjective visual improvement and satisfaction with quality-of-life improvement. The preoperative and 4-month postoperative quality of life were evaluated, and the association with the patients' characteristics, type of surgery performed, presence of systemic or ocular diseases, and subjective patient satisfaction was analyzed. RESULTS: The preoperative visual acuity in the operated eye had a low correlation (0.11) with the quality-of-life score; visual acuity in the better eye had a higher correlation (0.29). Quality-of-life improvement was moderately correlated with visual acuity improvement and patient satisfaction. Overall, quality-of-life scoring improved in all domains after surgery, with an overall effect size of 0.68. Quality-of-life scores improved postoperatively in 83.6% of patients, did not change in 3.6%, and were worse in 12.7%. Visual acuity improved in 94.5%, remained the same in 2.7%, and was worse in 2.7%. There was no difference in quality-of-life improvement between extracapsular cataract extraction and phacoemulsification at 4 months. Patients with systemic or ocular diseases and those who were phakic in the fellow eye had lower quality-of-life improvement; however, this was statistically significant for systemic diseases only. Age and sex did not affect quality of life or its improvement. CONCLUSIONS: The quality of life and surgical outcomes in cataract patients were assessed by a simple questionnaire. The local culture and environment should be taken into consideration in the questionnaire design. PMID- 14522295 TI - Effect of a fixed dorzolamide-timolol combination on intraocular pressure after small-incision cataract surgery with Viscoat. AB - PURPOSE: To evaluate the effect of a fixed dorzolamide-timolol combination (Cosopt) on intraocular pressure (IOP) after small-incision cataract surgery with sodium chondroitin sulfate 4%-sodium hyaluronate 3% (Viscoat). SETTING: Department of Ophthalmology, University of Vienna, Vienna, Austria. METHODS: This prospective randomized study comprised 76 eyes of 38 patients scheduled for small incision cataract surgery in both eyes. Patients were randomized to receive 1 drop of the fixed dorzolamide-timolol combination in 1 eye or no treatment (control) immediately after cataract surgery. The fellow eye received the other assigned treatment. Cataract surgery was performed with Viscoat in an identical fashion in both eyes. The IOP was measured preoperatively and 6 hours, 20 to 24 hours, and 1 week postoperatively. RESULTS: Six hours after surgery, the mean increase in IOP was significantly lower in the dorzolamide-timolol group than in the control group (4.3 mm Hg +/- 5.6 [SD] versus 8.4 +/- 6.1 mm Hg; P =.003). Two eyes in the dorzolamide-timolol group and 9 in the control group had IOP spikes of 30 mm Hg or higher (P =.022). Twenty to 24 hours after surgery, the mean IOP change was -2.6 +/- 3.3 mm Hg in the dorzolamide-timolol group and 1.5 +/- 3.2 mm Hg in the control group (P<.001). CONCLUSIONS: The fixed dorzolamide-timolol combination was effective in reducing IOP 6 hours and 20 to 24 hours after cataract surgery. However, it did not prevent Viscoat-induced IOP spikes of 30 mm Hg or higher. PMID- 14522297 TI - Postoperative astigmatism and rotational stability after artisan toric phakic intraocular lens implantation. AB - PURPOSE: To evaluate deviations in the axis (intended versus achieved) and postoperative astigmatism after implantation of an Artisan toric phakic intraocular lens (IOL). SETTING: University Eye Hospital, Mainz, Germany. METHODS: This prospective study comprised 29 eyes with high ametropia and astigmatism. All eyes had uneventful implantation of a toric phakic IOL through a superior scleral tunnel incision at 12 o'clock. After a minimum of 6 months, the uncorrected visual acuity (UCVA), best correct visual acuity, refraction, and astigmatism were analyzed in all eyes. A multivariate analysis of postoperative astigmatism was performed. RESULTS: After a follow-up of at least 6 months, 95% of eyes were within +/-1.00 diopter (D) of emmetropia and 85% of eyes has a UCVA of 20/30 or better. The difference between the mean intended cylinder axis and achieved cylinder axis was 3.9 degrees (median 3 degrees; range to 13 degrees). The difference between the mean intended axis and the achieved axis between miosis and mydriasis was 1.8 degrees (median 1.5 degrees; range 0 to 5 degrees). The mean postoperative astigmatism after 6 months was 0.56 D with an axis of 31 degrees. Doubled-angle scatterplot analysis showed a tendency toward more flattening in the vertical meridian. CONCLUSIONS: During the 6-month follow-up, no significant rotation was observed after implantation of Artisan toric phakic IOLs to correct high ametropia. A sutureless sclerocorneal superior approach for phakic IOL insertion resulted in moderate to low astigmatism. Induced astigmatism should be taken into consideration during preoperative planning. PMID- 14522298 TI - Aqueous humor levels of topically applied bupivacaine 0.75% in cataract surgery. AB - PURPOSE: To measure the intraocular levels of bupivacaine 0.75% topically applied before phacoemulsification and to develop standards for topical anesthesia in cataract surgery. SETTING: Department of Ophthalmology, University Hospitals of Leicester, Leicester, United Kingdom. METHODS: Forty eyes having phacoemulsification for senile cataract under topical anesthesia without sedation were randomly assigned to 1 of 2 preoperative topical anesthesia regimens. Bupivacaine 0.75% was applied in 0.1 mL drops 3 times in the 30 minutes before surgery in 18 eyes and 6 times in the 60 minutes before surgery in 22 eyes. Aqueous humor and serum samples were taken at the start of surgery and the bupivacaine levels measured. A visual analog pain score scale was used to indicate intraoperative pain. RESULTS: The mean aqueous humor level of bupivacaine was 5.9 microg/mL +/- 4.3 (SD) after 3 drops and 5.7 +/- 4.0 microg /mL after 6 drops. The blood levels were less than 1.0 microg/mL. There was no statistically significant difference in the intraocular level of bupivacaine between the 2 groups. There was no difference in the age or sex distribution between the 2 groups, although there was an increase in the intraocular level of bupivacaine with age (approximately 1.4 microg/mL per decade; P =.048). There was no clear pattern associating the pain score with age, sex, or intraocular level of bupivacaine. CONCLUSIONS: A 3-drop regimen of bupivacaine 0.75% in the half hour before cataract surgery penetrated the eye as effectively as 6 drops in the 1 hour before surgery and provided good analgesia for phacoemulsification. Bupivacaine 0.75% penetrated the eye increasingly effectively with increasing age. PMID- 14522299 TI - Aqueous humor levels of topically applied levofloxacin, norfloxacin, and lomefloxacin in the same human eyes. AB - PURPOSE: To assess the relative penetration of topical eyedrops of 3 fluoroquinolones into the aqueous humor in human eyes. SETTING: Department of Ophthalmology, Sano-Kosei Hospital, Sano, Japan. METHODS: Fifty-nine cataract patients (36 women, 23 men) received 3 drops each of levofloxacin 0.5%, norfloxacin 0.3%, and lomefloxacin 0.3% in the same eye at 15-minute intervals beginning 90 minutes before cataract surgery. At the beginning of surgery, 50 microL of aqueous humor was aspirated from the anterior chamber and stored at -80 degrees C until analyzed. The drug concentrations in the samples were analyzed using high-performance liquid chromatography. RESULTS: Five patients were excluded from the study because their sample volumes were insufficient. Norfloxacin was detected in 3 patients; the mean aqueous humor level was 0.10 microg/mL +/- 0.02 (SD). Levofloxacin was detected in all cases; the mean aqueous humor level was 0.60 +/- 0.28 microg/mL (n = 54). Lomefloxacin was not detected in 10 patients; the mean aqueous humor level was 0.23 +/- 0.11 microg/mL (n = 44). CONCLUSION: Topically applied levofloxacin had better penetration into the aqueous humor than lomefloxacin and norfloxacin. PMID- 14522300 TI - Reproducibility of morphoscopic contrast sensitivity testing with the Visual Capacity Analyzer. AB - PURPOSE: To determine the reproducibility of morphoscopic contrast sensitivity test values in healthy subjects using the Visual Capacity Analyzer (VCA) (L2 Informatique). SETTING: Institute of Vision Research, Department of Ophthalmology, Yong-Dong Severance Hospital, Yonsei University, Seoul, South Korea. METHODS: Five healthy volunteers were recruited for this study. With the VCA, 1 eye of each person was tested with different sized letters displayed on a computer screen at 11 spatial frequencies ranging from low (3.0 cycles/deg [cpd]) to high (30.0 cpd). The measurement was repeated 5 times under 2 screen luminance levels (maximum and 3 cd/m2). RESULTS: Under maximum luminance background, the coefficient of variation (CV) and reliability coefficient (RC) at the spatial frequencies examined ranged from 4.3% to 35.0% and 89.1% to 99.8%, respectively. Under a screen luminance of 3 cd/m2, the CV ranged from 0.5% to 15.9% and the RC, from 97.5% to 100.0%. CONCLUSION: At the spatial frequencies examined, morphoscopic contrast sensitivity testing using the VCA had a high level of reproducibility and may be useful in measuring a patient's visual function in the actual environment. PMID- 14522301 TI - Stress-strain measurements of human and porcine corneas after riboflavin ultraviolet-A-induced cross-linking. AB - PURPOSE: To evaluate the biomechanical effect of combined riboflavin-ultraviolet A (UVA) treatment on porcine and human corneas. SETTING: Department of Ophthalmology, Technical University of Dresden, Dresden, Germany. METHODS: Corneal strips from 5 human enucleated eyes and 20 porcine cadaver corneas were treated with the photosensitizer riboflavin and irradiated with 2 double UVA diodes (370 nm, irradiance = 3 mW/cm2) for 30 minutes. After cross-linking, static stress-strain measurements of the treated and untreated corneas were performed using a microcomputer-controlled biomaterial tester with a prestress of 5 x 10(3) Pa. RESULTS: There was a significant increase in corneal rigidity after cross-linking, indicated by a rise in stress in treated porcine corneas (by 71.9%) and human corneas (by 328.9%) and in Young's modulus by the factor 1.8 in porcine corneas and 4.5 in human corneas. The mean central corneal thickness was 850 microm +/- 70 (SD) in porcine corneas and 550 +/- 40 microm in human corneas. CONCLUSIONS: Riboflavin-UVA-induced collagen cross-linking led to an increase in mechanical rigidity in porcine corneas and an even greater increase in human corneas. As collagen cross-linking is maximal in the anterior 300 microm of the cornea, the greater stiffening effect in human corneas can be explained by the relatively larger portion of the cornea being cross-linked in the overall thinner human cornea. PMID- 14522302 TI - Endothelial cell damage after riboflavin-ultraviolet-A treatment in the rabbit. AB - PURPOSE: To evaluate the possible cytotoxic effect of combined riboflavin ultraviolet-A (UVA) treatment on the corneal endothelium. SETTING: Department of Ophthalmology, Technical University of Dresden, Dresden, Germany. METHODS: The right eyes of 34 New Zealand White rabbits were treated with riboflavin and various endothelial UVA doses ranging from 0.16 to 0.9 J/cm2 (0.09 to 0.5 mW/cm2, 370 nm) and postoperative enucleation times of 4 hours and 24 hours. The endothelial cells were evaluated in histological sections. The terminal deoxynulceotidyl transferase deoxy-UTP-nick-end labeling (TUNEL) technique and transmission electron microscopy were used to detect apoptosis. RESULTS: There was no endothelial damage in the 6 rabbit eyes enucleated at 4 hours. In those enucleated at 24 hours, there was significant necrosis and apoptosis of endothelial cells in the corneas treated with an endothelial dose of > or =0.65 J/cm2 (0.36 mW/cm2), which is about twice the endothelial UVA dose used in the treatment of keratoconus patients. CONCLUSIONS: In rabbit corneas with a corneal thickness less than 400 microm, the endothelial UVA dose reached a cytotoxic level of > or =0.65 J/cm2 (0.36 mW/cm2) using the standard surface UVA dose of 5.4 J/cm2 (3 mW/cm2). Pachymetry should be routinely performed before riboflavin UVA treatment; in thinner corneas, irradiation should not be done because of the cytotoxic risk to the endothelium. PMID- 14522303 TI - Intracameral antibiotic agents for endophthalmitis prophylaxis: a pharmacokinetic model. AB - PURPOSE: To assess in vitro whether intracameral antibiotic agents are plausibly effective prophylaxis against Staphylococcus aureus endophthalmitis. SETTING: University research laboratory.Staphylococcus aureus was cultured for 8 hours with vancomycin (50 microg/mL) or piperacillin/tazobactam (512/64 microg/mL). Without reducing the bacterial counts, the antibiotic concentration was halved every 2 hours to simulate the half-life of antibiotic agents in the anterior chamber. The number of colony forming units (CFU)/mL was measured at 0, 2, 4, 6, and 8 hours. RESULTS: Vancomycin inhibited the growth of methicillin-resistant S aureus such that the CFU/mL was 1000 times higher in the control group after 8 hours. Piperacillin/tazobactam nearly eliminated 1 oxacillin-susceptible strain and reduced the CFU/mL of another strain 1000-fold. CONCLUSIONS: Intracameral vancomycin may inhibit the growth of S aureus in the anterior chamber after surgery. Inhibiting the growth could increase the chances that the eye's innate defenses would eliminate the bacterial contamination that could cause endophthalmitis. PMID- 14522304 TI - Intraocular lens with reversibly adjustable optical power: pilot study of concept and safety. AB - PURPOSE: To evaluate whether it is technically feasible and safe to implant and adjust an intraocular lens (IOL) with reversibly adjustable refractive power designed to correct residual postoperative refractive error. SETTING: Animal study facility, Klinikum Mannheim, University of Heidelberg, Heidelberg, Germany. METHODS: An Acri.Tec AR-1 posterior chamber IOL (PC IOL) was implanted in pig cadaver eyes and in rabbit eyes after the crystalline lens was removed by phacoemulsification. The IOL was manipulated to change the refractive power. The outcome measures were the stability of the IOL during implantation, IOL positioning and rotation, the ability to move the adjustment device after the IOL had been implanted for several weeks, clinical signs of inflammation, and changes in the histopathologic appearance of the eye. RESULTS: Implantation and adjustment of refractive power were possible. The eyes healed normally. There was no difference between eyes with the AR-1 PC IOL and eyes with a control IOL in inflammatory reaction, corneal transparency, or histopathologic appearance. CONCLUSION: The AR-1 PC IOL was easy to implant and well tolerated in rabbit eyes. Surgical adjustment of the adjusting element was performed with little effort several weeks after implantation of the IOL. PMID- 14522306 TI - Pediatric cataract surgery and intraocular lens implantation: practice styles and preferences of the 2001 ASCRS and AAPOS memberships. AB - A survey of pediatric cataract surgery and intraocular lens (IOL) implantation practice patterns of adult and pediatric cataract surgeons was performed in October 2001. Questionnaires were distributed to the American Society of Cataract and Refractive Surgery and the American Association for Pediatric Ophthalmology and Strabismus. The overall return rate was 12.6% and 41.0%, respectively. Results show that pediatric cataract surgery with IOL implantation is being performed at a younger age than 8 years ago. Also, pediatric cataract surgery practice patterns are evolving in step with advances in adult surgery but with notable differences. PMID- 14522305 TI - ESCRS Binkhorst lecture 2002: Pseudophakic preservative maculopathy. AB - Many antiglaucoma eyedrops are reported to cause cystoid macular edema (CME) in aphakia and pseudophakia. We review 4 clinical and laboratory studies that compare the incidence of CME in early postoperative pseudophakia in eyes that received preserved latanoprost and timolol, nonpreserved timolol, and the preserved and nonpreserved vehicle for these drugs and looked at the morphological damage to cells and the changes in the indicators of cytokine and prostaglandin (PG) synthesis caused by latanoprost and timolol and the preservative benzalkonium chloride. Based on the findings of these studies, which indicate that the preservative causes increased synthesis of PGs and other substances and intensified postoperative inflammation, the term pseudophakic preservative maculopathy is proposed for CME caused by antiglaucoma eyedrops. PMID- 14522307 TI - Transient central retinal artery occlusion after posterior sub-Tenon's anesthesia. AB - Two cases of transient central retinal artery occlusion were observed preoperatively after uneventful sub-Tenon's infusion of local anesthetic for cataract surgery and intraocular lens implantation. In these eyes, the retinal circulation reperfused spontaneously before surgery and there were no visual sequelae. A third case was observed in an eye after strabismus surgery with sub Tenon's anesthesia. The patient was left with profound visual loss in this eye. The cause of this complication is unknown, but possible factors include mechanical pressure from the bolus of the anesthetic solution or localized vasoconstriction from the anesthetic, producing a decrease in ocular blood flow. Suggestions to avoid this problem include not inserting the cannula too posteriorly, not injecting forcibly against resistance, and using the minimum volume of anesthetic possible. PMID- 14522308 TI - Unexpected flap thickness in laser in situ keratomileusis. AB - We report a case of an unexpected thick flap during laser in situ keratomileusis (LASIK) that led to abandonment of surgery. This report illustrates the importance of stromal bed measurements after flap creation in LASIK. A thicker than-expected flap can lead to a thinner-than-anticipated residual cornea and subsequent ectasia or even perforation during laser ablation. It is possible that reports of ectasia in normal thickness corneas reflect thicker-than-anticipated flaps. PMID- 14522309 TI - Necrotizing scleritis after intraocular surgery associated with the use of polyester nonabsorbable sutures. AB - We present 2 elderly patients who developed necrotizing scleritis after cataract extraction in which the wound was sutured with polyester. In the first case, a 73 year-old woman who had right phacoemulsification via a limbal incision developed necrotizing scleritis 8 months after surgery. The eye became progressively painful and phthisical, necessitating enucleation. Microscopic examination of the enucleated globe showed a predominantly lymphocytic infiltration of the ocular tissues with no evidence of an infectious agent. In the second case, a 78-year old woman had bilateral extracapsular cataract extraction through a limbal incision closed with a polyester suture. The patient presented 3 years later with bilateral necrotizing sclerokeratitis. No underlying systemic vasculitis or autoimmune condition was identified in either patient. To our knowledge, the association of necrotizing scleritis after intraocular surgery and polyester fiber suture material (Mersilene) has not been described. PMID- 14522310 TI - Lateral tarsorrhaphy for prevention of postoperative complications resulting from globe luxation. AB - We present a case of spontaneous globe luxation and wound dehiscence that developed in a patient with proptosis 1 day after phacoemulsification with posterior chamber intraocular lens implantation. The wound was repaired and a lateral tarsorrhaphy was performed with successful prevention of further luxations. Upon cataract extraction in the patient's other eye, lateral tarsorrhaphy was performed concurrently to prevent operative complications. This report underscores the importance of lateral tarsorrhaphy performed at the time of cataract surgery to avoid postoperative globe luxation with possible wound dehiscence in patients with proptosis or shallow orbits and a history of luxation. PMID- 14522311 TI - Phacoemulsification and intraocular lens implantation in an Alport's syndrome patient with bilateral anterior and posterior lenticonus. AB - We report a case of intraocular lens (IOL) implantation with a continuous curvilinear capsulorhexis and phacoemulsification in a 30-year-old man manifesting anterior and posterior lenticonus. The surgery was uneventful and on follow-up, the IOL was well centered in the bag. PMID- 14522312 TI - Cataract surgery and intraocular lens implantation in a retinoblastoma case treated by external-beam radiation therapy. AB - We report a case of extracapsular cataract extraction with posterior chamber intraocular lens implantation in a pediatric patient with a radiation-induced cataract that developed after combined external-beam radiation therapy and chemotherapy for retinoblastoma. A 12-year follow-up revealed effective tumor regression and sustained visual recovery. PMID- 14522313 TI - Retained Descemet's membrane after penetrating keratoplasty. AB - Retained Descemet's membrane is a rare complication of penetrating keratoplasty (PKP). The membrane can become progressively opaque and threaten the health of the corneal graft. We report such a case involving a 53-year-old man. Simultaneous surgical removal of the membrane and phacoemulsification with intraocular lens implantation were performed 3 months after PKP. The graft remained clear and best corrected visual acuity was 20/20 2 years after secondary surgery. PMID- 14522314 TI - Traumatic tattooing of the skin and ocular surface. PMID- 14522315 TI - Potential uses of the Honan balloon in ophthalmic practice. PMID- 14522316 TI - Orbital regional anesthesia: let's be precise. PMID- 14522317 TI - The accuracy of sentinel lymph node biopsy in multicentric and multifocal invasive breast cancers. AB - BACKGROUND: Sentinel lymph node biopsy (SLNB) has proved to be an accurate alternative to complete axillary lymph node dissection (ALND) in clinically node negative breast cancer patients. Multicentric (MC) and multifocal (MF) invasive breast cancers are considered to be relative contraindications to SLNB. We examine the accuracy of SLNB in patients with MC and MF invasive breast cancers. STUDY DESIGN: From September 1996 to August 2001, a total of 3,501 patients with clinically node-negative breast cancer underwent SLNB using both blue dye and radioisotope at our institution. A total of 70 patients had MC or MF invasive breast cancer, a successful SLNB, and mastectomy for local control. All had >/=10 axillary nodes excised (including the SLN) in a planned ALND. Exclusion criteria included MC and MF in situ carcinoma; breast conservation; previous breast irradiation, ALND, or SLNB; recurrent breast cancer; neoadjuvant chemotherapy; or ALND based solely on SLNB pathologic examination. RESULTS; The incidence of axillary metastases was 54% (38 of 70). SLNB accuracy was 96% (67 of 70), sensitivity 92% (35 of 38), and false-negative rate 8% (3 of 38). All patients with an inaccurate SLNB had a dominant invasive tumor >5 cm and one patient had palpable axillary disease intraoperatively. The SLN was the only site of axillary metastasis in 37% (14 of 38). Results were compared with those of published SLNB validation studies, most of which reflect experience with single-site invasive breast cancers. No statistically significant difference was noted for accuracy, sensitivity, or false-negative rate. CONCLUSIONS: SLNB accuracy in MC and MF disease is comparable with that of published validation studies. MC and MF patients with a dominant T3 tumor (>5 cm) or axillary disease palpable intraoperatively should have a concurrent formal ALND. Our retrospective data suggest SLNB may be used as a reliable alternative to conventional ALND in selected patients with MC or MF disease. Further studies in this patient population are warranted. PMID- 14522318 TI - Laparoscopic gastric bypass versus laparoscopic adjustable gastric banding: a comparative study of 1,200 cases. AB - BACKGROUND: Indications for and results of laparoscopic adjustable gastric banding (LAGB) and laparoscopic gastric bypass (LGB) are still controversial, especially between Europe and the United States. The recent availability of gastric bandings in the United States made it necessary to compare the two techniques. STUDY DESIGN: We compared a series of 456 LGB to a series of 805 LAGB performed in two different institutions. Body mass index (BMI), complication rate, mortality, and excess weight loss (EWL) after 3, 6, 12, and 18 months were obtained. A Fischer's exact test and a Student t test with covariance analysis were used for statistical analysis. RESULTS: Results are expressed as a mean +/- standard deviation, comparing LGB with LAGB. Preoperative BMI was 49.4 +/- 8.3 kg/m(2) versus 42.2 +/- 4.9 kg/m(2) (p = 0.0001), respectively. Perioperative major complication rates were 2.0% versus 1.3% (NS), and the early postoperative major complication rates were 4.2% versus 1.7% (p = 0.02), respectively. Mortality rate was 0.4% versus 0% (NS), respectively. The global EWL was 36.3% for LGB versus 14.7% for LAGB at 3 months (p < 0.0001), 51.6% versus 21.9% at 6 months (p < 0.0001), 67.0% versus 33.3% at 12 months (p < 0.0001), and 74.6% versus 40.4% at 18 months (p < 0.0001), respectively. Longterm followup for the LAGB group showed an EWL of 47% at 2 years, 56% at 3 years, and 58% at 4 years. Patients were sorted after their preoperative BMI (30 to 40, 40 to 50, and 50 to 60 kg/m(2)). The EWL at 3, 6, 12, and 18 months was statistically superior in the LGB group, for any BMI ranges. CONCLUSIONS: These data suggest that LGB provides a higher EWL at 18 months, compared with LAGB, and this was true for any preoperative BMI range. It is associated with a higher early postoperative complication rate. PMID- 14522321 TI - Factors associated with operative outcomes in laparoscopic gastric bypass. AB - BACKGROUND: Laparoscopic gastric bypass (GBP) is becoming a common approach for treatment of morbid obesity. We analyzed preoperative factors that may be associated with operative outcomes in laparoscopic GBP. STUDY DESIGN: This prospective study evaluates 150 consecutive laparoscopic GBP procedures performed by a single surgeon. Preoperative factors were grouped into three categories: 1) patient-specific (gender, age, abdominal surgical history, smoking), 2) obesity specific (body mass index, hypertension, diabetes, sleep apnea), and 3) procedure specific (operative experience of the surgeon [75 cases or less versus more than 75 cases]). Length of operation (240 minutes or less versus more than 240 minutes), postoperative complications (yes versus no), major complications (yes versus no), reoperation (yes versus no), and length of hospital stay (4 days or less versus more than 4 days) were the operative outcomes considered. In this series all patients who had a major complication required a reoperation. Data were analyzed using univariate and multiple logistic regression analyses. RESULTS: Operative experience of surgeon (75 cases or less) was associated with lengthy operative time (adjusted odds ratio [AOR], 3.8; 95% confidence interval [CI], 1.7 to 8.3), major complications (AOR, 15.0; 95% CI, 1.5 to 143.0), and a lengthy (more than 4 days) hospital stay (AOR, 4.5; 95% CI, 1.1 to 18.0). Higher patient age (50 years or more) was associated with more postoperative complications (AOR, 11.4; 95% CI, 3.0 to 43.1) and major complications (AOR, 7.6; 95% CI, 1.1 to 48.7). Male gender also was associated with more postoperative complications (AOR 5.2; 95% CI, 1.1 to 23.1). Obesity-related comorbidities, body mass index, past abdominal surgical history, and smoking had no statistical association with operative outcomes in this study. CONCLUSIONS: There is an association of clinical outcomes after laparoscopic GBP with the age and gender of the patient and the operative experience of the surgeon. An operative experience of more than 75 laparoscopic GBP cases was associated with decreases in operative time, length of hospital stay, and number of major complications. PMID- 14522324 TI - Increasing esophageal length: a comparison of laparoscopic versus transthoracic esophageal mobilization with and without vagal trunk division in pigs. AB - BACKGROUND: New laparoscopic techniques allow both mediastinal mobilization and performance of a Collis gastroplasty when necessary, and the utility of a transthoracic approach is questioned. The aim of this study was to compare the increase in esophageal length achievable with laparoscopic and transthoracic esophageal mobilization in pigs, and to assess the impact of vagal trunk division on esophageal length. STUDY DESIGN: Baseline esophageal length was obtained in 20 farm pigs by measuring the distance between a stitch placed in the esophagus to a K-wire placed in a vertebral body. Subsequently, laparoscopic and then transthoracic mediastinal mobilization of the esophagus were performed in 15 pigs and the length gain after each procedure recorded. In 7 of 15 animals, the vagal nerve trunks were divided after esophageal mobilization and the increase in esophageal length measured. In five animals, vagal trunk division was performed without earlier esophageal mobilization. RESULTS: Esophageal length gain after laparoscopic mobilization (median 4 mm) was significantly less than that after transthoracic mobilization (median 12 mm, p < 0.0001). Unilateral vagal nerve transection resulted in a median 2.5 mm of esophageal length gain compared with a median of 6.25 mm with division of both vagal trunks. Maximal esophageal lengthening (median 18.5 mm) occurred with a combination of esophageal mobilization and bilateral vagal trunk division. CONCLUSIONS: Esophageal length gain after transthoracic mobilization in normal pigs is significant, and would likely be even greater in patients with gastroesophageal reflux disease with concomitant mediastinal inflammation. Transthoracic mobilization alone likely will allow successful reduction of the gastroesophageal junction below the diaphragm in many patients who might otherwise require a Collis gastroplasty. PMID- 14522325 TI - Radical oophorectomy with primary stapled colorectal anastomosis for resection of locally advanced epithelial ovarian cancer. AB - BACKGROUND: The aim of this study was to describe the feasibility, associated morbidity, and efficacy of radical oophorectomy with primary stapled colorectal anastomosis among patients with locally advanced ovarian cancer with contiguous extension to or encasement of the reproductive organs, pelvic peritoneum, cul-de sac, and sigmoid colon. STUDY DESIGN: Thirty-one consecutive patients undergoing radical oophorectomy as part of an initial maximal surgical effort for International Federation of Obstetrics and Gynecology (FIGO) stage IIIB-IV ovarian cancer were prospectively collected from October 1, 1997 through November 30, 2001. The surgical technique, associated morbidity, and clinical outcomes are described. RESULTS: The median age was 63 years. All patients had advanced-stage epithelial ovarian cancer: FIGO stage IIIB (6.5%), stage IIIC (64.5%), stage IV (29.0%). Median operating time was 240 minutes (range 165 to 330 minutes), and the median estimated blood loss was 700 mL (range 300 to 2,900 mL). All patients underwent en bloc rectosigmoid colectomy with primary stapled anastomosis without protective intestinal diversion. There was one (3.2%) anastomotic breakdown requiring reoperation and colostomy. Complete clearance of macroscopic pelvic disease was achieved in all cases. Overall, 87.1% of patients were left with optimal ( 10% area) in 50% of high- and 22% of low-ligation control animals. DCA treatment eliminated significant muscle necrosis in 100% of high-ligation animals. Muscle histology was similar in control and DCA-treated low-ligation animals. CONCLUSIONS: Treatment with DCA during acute arterial occlusion did significantly lower markers of anaerobic metabolism and reduced muscle necrosis in a rabbit model of acute hind-limb ischemia. DCA delivery through collateral blood flow may prolong the ischemia time interval before the onset of irreversible muscle injury and potential limb loss. PMID- 14522329 TI - Consequences of delayed diagnoses in trauma patients: a prospective study. AB - BACKGROUND: The approach to trauma care has improved in recent decades but delayed diagnoses still occur. This study aimed to analyze the prevalence and consequences of delayed diagnoses in a single European trauma center. The effect of a systematic reexamination of the patient (tertiary survey) and reevaluation of x-rays and CT scans was evaluated. STUDY DESIGN: We prospectively registered complications among all trauma patients admitted to our hospital from January 1, 1996, to January 1, 2000. All relevant trauma and patient-related data were added by the physician to a hospital-wide trauma database with client server architecture. Complications including delay in diagnosis were subsequently added to this database. Admitted trauma patients underwent a tertiary survey and all x rays and CT scans were reevaluated within 24 hours after admission. RESULTS: A total of 3,879 patients were studied and 1,016 complications were registered. Of all complications 55 concerned delayed diagnoses detected in 49 patients (1.3%). In 28 of these patients (57.1%) the tertiary survey (20 of 49; 40.8%) and reevaluation of x-rays and CT scans (8 of 49; 16.3%) resulted in detection of delayed diagnoses within 24 hours. Detection of the remaining 21 delayed diagnoses occurred after more than 24 hours. Delayed diagnoses resulted in delayed treatment in 27 of the 49 patients (55.1%) and surgery was necessary in 12 patients (24.5%). None of the delayed diagnoses resulted in death. CONCLUSIONS: A prospective trauma and complication registration enables evaluation of the delays in diagnosis. In our study population more than half of the delayed diagnoses could be detected by a tertiary survey and reevaluation of x-rays and CT scans. Attempts to decrease the number of delayed diagnoses should prevent delays in treatment and improve the quality of trauma care. PMID- 14522330 TI - Professionalism in surgery. PMID- 14522331 TI - Historical perspective on medical errors: Richard Cabot and the Institute of Medicine. PMID- 14522332 TI - What's new in gynecology and obstetrics. PMID- 14522333 TI - A rural, community-based program can train surgical residents in advanced laparoscopy. AB - BACKGROUND: Advanced laparoscopy requires mastery of complex surgical skills. A steep learning curve, lack of an adequate number of cases, and a shortage of experienced staff are reasons cited as barriers to the acquisition of these skills by surgical residents. We hypothesize that advanced laparoscopy can be taught during residency without additional fellowship training. STUDY DESIGN: ast surgical residents who completed training at our rural, community-based, 140-bed hospital from 1992 to 2000 were contacted by mailed surveys and a followup telephone interview. Advanced laparoscopy was defined as cases other than cholecystectomy, appendectomy, and diagnostic laparoscopy. Five attending surgeons routinely perform advanced laparoscopy. RESULTS: The response rate to the survey was 93.3% with 15 of 18 graduates currently practicing general surgery and 100% of the surgeons performing advanced laparoscopy. Laparoscopic herniorrhaphy, splenectomy, colectomy, Nissen fundoplication, and adrenalectomy were performed by 12 (85.7%), 10 (71.4%), 11 (78.6%), 13 (92.9%), and 9 (64.3%) surgeons, respectively. Eight (57.1%) surgeons reported confidence to perform advanced laparoscopy immediately after residency. All graduating chief residents from the last 3 years expressed this confidence. On average each of two chief residents from the past 3 academic years graduated with 99 basic and 50 advanced laparoscopic cases. CONCLUSIONS: A rural, community-based program can train residents to perform advanced laparoscopy. Increasing the volume of advanced cases handled by resident correlates with increasing confidence in graduates. PMID- 14522334 TI - Work hours reform: perceptions and desires of contemporary surgical residents. AB - BACKGROUND: New Accreditation Council for Graduate Medical Education (ACGME) requirements on resident duty hours are scheduled to undergo nationwide implementation in July 2003. General surgery residents, because of their long duty hours, are likely to be among those most affected by changes imposed to comply with the ACGME requirements. There are few contemporary data on their attitudes toward work hours reform. STUDY DESIGN: The study entailed a region wide survey of residents enrolled in general surgery residencies in New England to characterize the perceptions and desires of surgical residents on the issue of work hours reform. RESULTS: Respondents reported working a mean of 105 +/- 0.7 hours per week, considerably more than the 80-hour limit stipulated by the ACGME. Of the respondents, 81% reported that sleep deprivation had negatively affected their work. A strong majority of respondents believe that work hours reform would improve their quality of life but less than one half expect it to have a positive impact on patient care. A greater percentage of senior residents than junior residents (p < 0.05) have negative perceptions of work hour limitations, particularly with respect to consequences for patient care. Other findings suggest that residents who have actually experienced work hour restrictions are less positive about such restrictions than these residents who had not yet experienced them. CONCLUSIONS: Changes imposed by residency programs to comply with work hour requirements might have detrimental effects on senior residents and patient care. The impact of such changes should be carefully monitored as the ACGME requirements are implemented. PMID- 14522336 TI - Liver regeneration. PMID- 14522337 TI - Retroperitoneal hemangiopericytoma. PMID- 14522338 TI - Radial choroidal ruptures in sclopetaria. PMID- 14522339 TI - Portless laparoscopic stay-suture. PMID- 14522340 TI - Robotic revelation: laparoscopic radical prostatectomy by a nonlaparoscopic surgeon. PMID- 14522341 TI - Evaluation of pathologic nipple discharge with ductoscopy. PMID- 14522342 TI - Inconsistent finding of tachycardia in World War II combat casualties. PMID- 14522343 TI - Fresh frozen plasma use in liver resection. PMID- 14522345 TI - Extramammary Paget's disease of the vulva. PMID- 14522346 TI - Inguinal hernia management. PMID- 14522349 TI - [Absorbed dose evaluation of thyroid during nasopharynx and breast carcinoma irradiation by in vivo dosimetry]. AB - AIMS: The thyroid dysfunction after radiotherapy has led to evaluate the dose received by thyroid during nasopharynx and breast carcinoma irradiation. This evaluation was facilitated by in vivo dosimetry. The aims of this work were to evaluate the thyroid dose and to compare released dose at the reference point in the two localizations. PATIENTS AND METHODS: A total of 30 patients were evaluated: 18 patients with nasopharynx carcinoma and 12 patients with breast carcinoma were included in the final analysis. In the first group, the total thyroid but the isthmus was irradiated. On the other hand, in the second group, only one thyroid lobe was included into the target volume. All patients have been treated by gamma rays of cobalt 60 and the thyroid absorbed dose was measured by semiconductor dosimeters (Scanditronix DPD6). These dosimeters were calibrated in the same geometric conditions of the irradiation. RESULTS: The measured absorbed dose of the thyroid parenchyma was equal to the calculated absorbed dose at the target that has been specified at a depth of -3 cm. Under the block, the isthmus received 5.1 +/- 0.9 Gy (9.9 +/- 1.8%) in nasopharynx carcinoma irradiation. However, in breast carcinoma irradiation, the distal thyroid lobe and the isthmus received 2.9 +/- 0.7 Gy (6.55 +/- 1.56%) and 3.69 +/- 0.77 (8.39 +/- 1.76%), respectively. CONCLUSIONS: This study shows that the thyroid received a dose equal to the prescribed dose in both nasopharynx and breast cancer patients. It is recommended to follow the function of the thyroid gland in these patients. PMID- 14522350 TI - [Second malignancies following Hodgkin's disease treatment in Tunisia. Retrospective study of 26 cases observed at the institute Salah-Azaiz]. AB - PURPOSE: To collect second cancers in patients treated for Hodgkin disease (HD) during adolescence and young adulthood at Salah Azaiz Institute of Tunis. METHODS AND PATIENTS: We consider as second cancer all tumours other than HD observed in patients after treatment for HD. RESULTS: Twenty-five patients among 614 treated for HD between 1975 and 1991 developed 26 secondary tumours (4.2%). There were 17 males and eight females (sex ratio 2:1). Mean age at the diagnosis of HD was 32.5 years (12-56). HD was stage II (eight cases), stage III (14) and stage IV in three. The first treatment was combined chemotherapy and radiotherapy in 22 cases and only chemotherapy in three cases (stage IV). Radiotherapy was delivered with Cobalt 60 by large fields. Mean dose was 41.3 Gy (2 Gy/fraction in 21 and 3.3 in one). Chemotherapy was MOPP (13), MOPP and vinblastine (four), MOPP-ABVD (five), ABVD (two) and vinblastine only in one. Mean delay of second tumours was 114.5 months (40-276). There was five acute myeloid leukaemia, two digestive non Hodgkin lymphomas, five nodal high-grade lymphomas, three breast cancers (one in man associated with thyroid cancer), five lung cancers (three non-small cell and two of small cell type), two gastric tumours and one rectal cancer, one synovialosarcoma of the knee and one malignant Schwannoma of the neck. Median survival was 12.5 months (2-48). Twenty-one patients died and four are still alive with 8, 12, 24 and 48 months of follow-up. CONCLUSION: Second cancer risk after treatment for HD is not low. Risk factors and preventive strategies are discussed. PMID- 14522351 TI - [Epstein-Barr virus associated central nervous system leiomyosarcoma occurring after renal transplantation: case report and review of the literature]. AB - Central nervous system leiomyosarcomas are extremely rare, however, they became more frequent among immunodeficient patients, either in a patients infected with human immunodeficiency virus (HIV), or after organ transplantation. The data of the literature indicate that the infection by Epstein-Barr virus (EBV) plays a causal role in the development of these tumours but its precise role in the oncogenesis remains unresolved. We report a new case of EBV associated leiomyosarcoma of the left cavernous sinus occurring after renal transplantation. The epidemiological, clinical, pathological and therapeutic characteristics of these tumours are discussed. PMID- 14522352 TI - [Angiocentric lymphoma of the face: report of the 2 cases]. AB - Angiocentric lymphoma of the face is a rare malignancy. During 11 years, only two patients were treated for a lymphoma of the face. The first patient, 31-year-old, had an angiocentric lymphoma of the nasal cavity, the second patient, 40-year old, had an angiocentric lymphoma of the nasopharynx and the maxillary sinuses, with an extension the hard palate. According to the Ann Arbor classification, the disease was stage IEab and stage IIEbb, respectively. The first patient was treated by chemotherapy alone (COP regimen) and the second patient underwent combined chemotherapy (CHOP) and radiotherapy. The follow-up was marked by progression of the disease 7 and 9 months after the end of the treatment in the two cases. For the angiocentric lymphoma, treatment based on radiotherapy or chemotherapy alone or combined does not often provide local control and the overall survival rate does not exceed 40% at 5 years. PMID- 14522353 TI - Cerebellous metastases in patients with uterine cervical cancer. Two cases reports and review of the literature. AB - Brain metastases from cervical cancer are extremely rare. We report on two patients who developed cerebellous metastases following uterine cervical cancer. The interval between diagnosis of the primary cancer and diagnosis of brain metastasis was 8 months. The main complaint was symptoms of increased intracranial pressure and cerebellous syndrome. Surgical excision of the brain lesion followed by radiation therapy was performed in the first case. The second patient received palliative radiation therapy. The first patient died 8 months after diagnosis. The second patient is alive 2 months after diagnosis. PMID- 14522354 TI - [Treatment with charged particles beams: hadrontherapy part I: physical basis and clinical experience of treatment with protons]. AB - Protons have physical characteristics, which differ from those of photons used in conventional radiotherapy. Better shielding of critical organs is obtained by using their particular ballistic (Bragg peak and lateral narrow penumbra). Some indications as ocular melanoma, chordoma and chondrosarcoma of the base of skull are now strongly accepted by the radiation oncologist community. Others are still in evaluation: meningioma, locally advanced nasopharynx tumor and paediatric tumors. The aim of this review is to present the clinical results of a technic which seems "confidential" because of the rarety and the cost of equipments. PMID- 14522355 TI - [Treatment with neutrons: hadrontherapy part II: physical basis and clinical experience]. AB - Neutrons have radiobiological characteristics, which differ from those of conventional radiotherapy beams (photons) and which offer a theoretical advantage over photons to fight radioresistance by the differential relative biological effect of them between normal and tumour tissues. Neutron therapy beneficed of great interest between 1975 and 1985. Many of phase III trials were conducted and indications have been definitively deducted of them. After briefly describing the properties of neutron beams, this review discusses the indication of neutron therapy on the basis of the clinical results. Salivary, prostate tumours and sarcomas are the main indications of neutron therapy. In concern to the prostate cancers, other alternative treatments reduce the neutron therapy field. For sarcomas, the lack of randomised trials limits the impact of the interest of neutrons. For other tumours, the ratio benefice/risk of neutron therapy is inferior to these obtained with photons and they could not be considered like classical indications. PMID- 14522356 TI - [Volume-effect and radiotherapy [II]. Part II: volume-effect and normal tissue]. AB - The first part of our work has focused on the relationship between tumor volume and tumor control. Indeed, it is well known that the importance of irradiated volume could be a main parameter of radiation-induced complications. Numerous mathematical models have described the correlation between the irradiated volume and the risk of adverse effects. These models should predict the complication rate of each treatment planning. At the present time late effects have been the most studied. In this report we firstly propose a review of different mathematical models described for volume effect. Secondly, we will discuss whether these theoretical considerations can influence our view of radiation treatment planning optimization. PMID- 14522357 TI - Serofendic acid prevents acute glutamate neurotoxicity in cultured cortical neurons. AB - We have previously reported that a novel neuroprotective substance named serofendic acid was purified and isolated from ether extract of fetal calf serum. In the present study, we investigated the effect of serofendic acid on acute neurotoxicity induced by L-glutamate (Glu) using primary cultures of rat cortical neurons. Exposure of cortical cultures to Glu for 1 h caused a marked decrease in cell viability, as determined by trypan blue exclusion. This acute Glu neurotoxicity was prevented by N-methyl-D-aspartate (NMDA) receptor antagonists, extracellular Ca(2+) removal, nitric oxide (NO) synthase inhibitor and NO scavenger. Serofendic acid prevented acute Glu neurotoxicity in a concentration dependent manner. Acute neurotoxicity was induced by ionomycin, a Ca(2+) ionophore, and S-nitroso-L-cysteine, an NO donor. Serofendic acid also prevented both ionomycin- and S-nitroso-L-cysteine-induced neurotoxicity. Moreover, the protective effect of serofendic acid on acute Glu neurotoxicity was not affected by cycloheximide, a protein synthesis inhibitor, and actinomycin D, an RNA synthesis inhibitor. These results indicate that serofendic acid protects cultured cortical neurons from acute Glu neurotoxicity by reducing the cytotoxic action of NO and de novo protein synthesis is not required for this neuroprotection. PMID- 14522358 TI - Buspirone differentially modifies short-term memory function in a combined delayed matching/non-matching to position task. AB - This study investigated the action of 5-hydroxytryptamine (5-HT) mimetics on short-term memory function. The objective was to determine whether two closely related tasks could differentiate between partial 5-HT(1A) receptor activation, full 5-HT(1A) receptor activation and generalised enhanced serotonin (5-HT) activity. Male hooded Lister rats were trained to perform an operant-based combined delayed matching/non-matching to position task. Drugs used were: fluoxetine (3 mg/kg, i.p.), a selective 5-HT reuptake inhibitor; the full 5 HT(1A) receptor agonist, 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT; 0.3 mg/kg, s.c.); and the partial 5-HT(1A) receptor agonist, buspirone (1 mg/kg, i.p.). Buspirone differentially disrupted response accuracy depending on the style of trial. There was no such difference in the case of 8-OH-DPAT, which impaired accuracy in both delayed matching/non-matching to position task, while fluoxetine affected neither. Thus, the findings suggest that partial 5-HT(1A) receptor activation compromises cognitive function to a greater extent than full 5-HT(1A) receptor activation, although a dopaminergic component cannot be excluded since buspirone possesses some dopamine D2 receptor antagonist activity. Furthermore, it suggests that there is a differential role for 5-HT in these two closely related behavioural tasks. PMID- 14522359 TI - SR141716A, a cannabinoid CB1 receptor antagonist, improves memory in a delayed radial maze task. AB - An endogenous cannabinoid system may play an important role in controlling memory processes. SR141716A (N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4 dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamidehydrochloride), a selective cannabinoid CB(1) receptor antagonist, was studied in an eight-arm radial maze task in which either deficits or improvements in memory could be detected. This task required well-trained rats to recall after either a relatively short (3 h) or long (7 h) delay period where they had received rewards during an information phase in order to obtain the remaining rewards during a retention phase. SR141716A was administered intraperitoneally immediately after the information phase in order to determine the drug's effects on memory consolidation. Although SR141716A had no effect on the number of errors committed after a short interval, SR141716A significantly reduced the number of errors that occurred after 7 h. These results suggest that a cannabinoid CB(1) receptor antagonist can improve consolidation processes and thus may be useful in treating memory disorders. PMID- 14522360 TI - Amnesia induced by beta-amyloid fragments is counteracted by cannabinoid CB1 receptor blockade. AB - Administration of drugs activating cannabinoid CB(1) receptors in the brain induces memory deficit in rodents, and blockade of these receptors may restore memory capacity in these animals. Central administration of beta-amyloid or beta amyloid fragments may also lead to memory disturbances. This study was undertaken to study the involvement of cannabinoid CB(1) receptors in amnesia induced by beta-amyloid fragments in mice tested in a step-through passive avoidance paradigm. Pre-training intracerebroventricular (i.c.v.) injection of beta-amyloid fragments, beta-amyloid peptide-(25-35) (4, 8 or 16 nmol/mouse) or beta-amyloid peptide-(1-42) (200, 400, 800 pmol/mouse) 7 days prior to the learning trial reduced in a dose-dependent manner the retention of passive avoidance response. This effect was observed in two retention tests, 1 and 7 days after the learning trial. The two beta-amyloid fragments showed similar potency in reducing retention of passive avoidance behavior. This effect was counteracted by a single intraperitoneal (i.p.) injection of the cannabinoid CB(1) receptor antagonist, N (piperidin-l-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3 carboxamide hydrochloride (SR141716A, 1 mg/kg), made 30 min prior to the second retention test. The injection of SR141716A per se did not affect memory capacity of mice. The i.c.v. administration of beta-amyloid peptide-(25-35) (8 nmol/mouse) or of beta-amyloid peptide-(1-42) (400 pmol/mouse) made 30 min prior to the learning trial failed to affect the retention capacity of mice as measured 1 and 7 days later. Also, the i.p. injection of SR 141716A (1 mg/kg) made 30 min prior to the learning trial did not influence the behavioral response of mice injected with beta-amyloid peptide-(25-35) (8 nmol/mouse) or of beta-amyloid peptide-(1 42) (400 pmol/mouse) 7 days prior to the learning trial. These results show that beta-amyloid fragments induce a dose-dependent memory deficit. Their effect on memory retention depends upon the time of administration and seems to involve cannabinoid CB(1) receptors in the brain. PMID- 14522361 TI - Further evidence that melanocortins prevent myocardial reperfusion injury by activating melanocortin MC3 receptors. AB - In rats subjected to myocardial ischemia/reperfusion, melanocortin peptides, including gamma(1)-melanocyte-stimulating hormone (gamma(1)-MSH), are able to exert a protective effect by stimulating brain melanocortin MC(3) receptors. A non-melanocortin receptor belonging to a group of receptors for Phe-Met-Arg-Phe NH(2) (FMRFamide)-like peptides may be involved in some of the cardiovascular effects of the gamma-MSHs. FMRFamide-like peptides and gamma(1)-/gamma(2)-MSH share, among other things, the C-terminal Arg-Phe sequence, which seems to be essential for cardiovascular effects in normal animals. So we aimed to further investigate which receptor and which structure are involved in the protective effects of melanocortins in anesthetized rats subjected to myocardial ischemia by ligature of the left anterior descending coronary artery (5 min), followed by reperfusion. In saline-treated rats, reperfusion induced, within a few seconds, a high incidence of ventricular tachycardia and ventricular fibrillation, and a high percentage of death within the 5 min of observation period. Reperfusion was associated with a massive increase in free radical blood levels and with an abrupt and marked fall in systemic arterial pressure. The i.v. treatment (162 nmol/kg) during the ischemic period with the adrenocorticotropin fragment 1-24 [ACTH-(1-24): the reference protective melanocortin which binds all melanocortin receptors], as well as with both the melanocortin MC(3) receptor agonists gamma(2)-MSH and [D-Trp(8)]gamma(2)-MSH, reduced the incidence of ventricular tachycardia, ventricular fibrillation and death, the increase in free radical blood levels and the fall in arterial pressure. On the contrary, gamma(2)-MSH-(6 12) (a fragment unable to bind melanocortin receptors) was ineffective. Such protective effect was prevented by the melanocortin MC(3)/MC(4) receptor antagonist SHU 9119. In normal (i.e., not subjected to myocardial ischemia/reperfusion) rats, the same i.v. dose (162 nmol/kg) of gamma(2)-MSH, [D Trp(8)]gamma(2)-MSH and gamma(2)-MSH-(6-12) provoked a prompt and transient increase in arterial pressure; on the other hand, ACTH-(1-24), which lacks the C terminal Arg-Phe sequence, decreased arterial pressure, but only at higher doses. Heart rate of normal rats was not affected by any of the assayed peptides. The present data confirm and extend our previous findings that melanocortins prevent myocardial reperfusion injury by activating melanocortin MC(3) receptors. Moreover, they further support the notion that, in normal rats, cardiovascular effects of gamma-MSHs are mediated by receptors for FMRFamide-like peptides, for whose activation, but not for that of melanocortin MC(3) receptors, the C terminal Arg-Phe structure being relevant. PMID- 14522362 TI - Protection from myocardial stunning by ischaemia and hypoxia with the adenosine A3 receptor agonist, IB-MECA. AB - Guinea pig isolated working hearts were exposed to 30-min ischaemia by reducing coronary flow to 10%, followed by reperfusion. Aortic output fell to 4.5+/-4.5% of the pre-ischaemic value at reperfusion, recovering to 48.2+/-14.6% at 20-min post-reperfusion; the index of myocardial stunning. IB-MECA (N(6)-(3 iodobenzyl)adenosine-5'-N-methyluronamide, 3 x 10(-7) M), infused from 10 min into ischaemia, did not affect recovery of aortic output 20 min after reperfusion (41.9+/-1.9%). IB-MECA infused at reperfusion, however, significantly protected against stunning, aortic output recovering to 79.6+/-3.9% at 20-min post reperfusion. Hypoxic gassing (5% CO(2) in nitrogen, 30 min) of guinea pig isolated paced left atria and papillary muscles reduced the developed tension, recovering to 75% 5 min after re-oxygenation. This myocardial stunning was unaffected by IB-MECA (3 x 10(-7) M) added 10 min into hypoxia. IB-MECA added at reoxygenation significantly improved recovery, which was prevented by the adenosine A(3) receptor antagonist, 1-propyl-3-(3-iodo-4-aminobenzyl)-8-(4 oxyacetate)phenylxanthine (I-ABOPX, 1 x 10(-5) M). Thus, stimulation of adenosine A(3) receptors at reperfusion/reoxygenation in guinea pig cardiac preparations protects against myocardial stunning. PMID- 14522363 TI - Action of prostanoids on the emetic reflex of Suncus murinus (the house musk shrew). AB - Several prostanoids were investigated for a potential to induce emesis in Suncus murinus. The TP receptor agonist 11alpha,9alpha-epoxymethano-15S-hydroxyprosta 5Z,13E-dienoic acid (U46619) induced emesis at doses as low as 3 microg/kg, i.p. but the DP receptor agonist 5-(6-Carboxyhexyl)-1-(3-cyclohexyl-3-hydroxypropyl) hydantoin (BW245C) was approximately 1000 times less potent. The emetic action of U46619 (300 microg/kg, i.p.) was antagonized significantly by the TP receptor antagonist, vapiprost (P<0.05). EP (prostaglandin E(2), 17-phenyl-omega-trinor prostaglandin E(2), misoprostol and sulprostone), FP (prostaglandin F(2alpha) and fluprostenol) and IP (iloprost and cicaprost) receptor agonists failed to induce consistent emesis at doses up to 300-1000 microg/kg, i.p. Fluprostenol reduced nicotine (5 mg/kg, s.c.)-but not copper sulphate (120 mg/kg, intragastric) induced emesis; the other inconsistently emetic prostanoids were inactive to modify drug-induced emesis. The results indicate an involvement of TP and possibly DP and FP receptors in the emetic reflex of S. murinus. PMID- 14522365 TI - Inhibitory capacity of different steroids on neutrophil migration across a bilayer of endothelial and bronchial epithelial cells. AB - Neutrophil infiltration to the airway lumen is a common feature of respiratory inflammatory processes. The aim of this study was to evaluate whether different corticosteroids exert any selective effect on the migration of isolated neutrophils. A bilayer of cultured human endothelial and bronchial epithelial cells was used as a model for neutrophil migration through the blood-air barrier. Low spontaneous migration of neutrophils (2.8+/-0.9%, n=8; mean+/-S.E.M.) occurred, while in the absence of any steroid, a migration of 28.5+/-7.6% could be induced by lipopolysaccharide. Pre-incubation during 1 h of epithelial cells with dexamethasone, budesonide, or prednisolone (10(-10)-10(-4) M) showed in all instances a concentration-dependent inhibition following a bell-shaped curve. At 10(-7) M, both dexamethasone and budesonide were on the minimum effect peak of the bell-shaped curve. The peak for prednisolone was found at 10(-8) M. However, when steroid pre-incubation was extended to 4 h, a sigmoid curve was observed, with significant inhibition of migration at concentrations >10(-7) M. Steroids can inhibit neutrophil recruitment through two different pathways with distinct result, depending on the length of incubation time. PMID- 14522364 TI - Contribution of tachykinin receptor subtypes to micturition reflex in guinea pigs. AB - The aim of the present study was to determine the role of tachykinin in the micturition reflex in guinea pigs. We investigated the effects of tachykinin NK(1) receptor antagonists, GR205171 ([2-methoxy-5-(5-trifluoromethyl-tetrazol-1 yl)-benzyl]-(2S-phenyl-piperidin-3S-yl)-amine), CP99994 ((+), (2R, 3R)-3-(2 methoxybenzyl-amino)-2-phenylpiperidine) and FK888 (N(2)-[(4R)-4-hydroxy-1-(1 methyl-1H-indol-3-yl) carbonyl-L-prolyl]-N-methyl-N-phenylmethyl-3-(2-naphthyl)-L alaninamide), the tachykinin NK(2) receptor antagonist, SR48968 ((+)-N-methyl-[4 (4-acetylamino-4-phenyl piperidino)-2-(3, 4-dichloro-phenyl)butyl] benzamide), and the tachykinin NK(3) receptor antagonist, SB223412 ((S)-(-)-N-(alpha ethylbenzyl)-3-hydroxy-2-phenylquinoline-4-carboxamide) on rhythmic bladder contraction. GR205171 and CP99994 but not SR48968 or SB223412 reduced bladder contraction frequency. FK888 inhibited the frequency very slightly at the highest dose tested. The distribution of tachykinin NK(1) receptor antagonists to the central nervous system after intravenous administration was examined using an ex vivo binding assay. GR205171 was distributed to the brain and spinal cord, but the tachykinin NK(1) receptor antagonist, FK888, was not. These results suggest that tachykinin NK(1) receptors, which are located in the central nervous system, play an important role in micturition in guinea pigs. PMID- 14522366 TI - Effect of enoxaparin on high glucose-induced activation of endothelial cells. AB - Clinical and experimental studies indicate that low-molecular-weight heparins reduce inflammation. To uncover the possible mechanisms involved, we investigated the effect of a low-molecular-weight heparin, enoxaparin, on high glucose-induced activation of endothelial cells. Bovine valvular endothelial cells and human endothelial cell line, EA hy926 were cultured in medium containing 5 mM (normal glucose) or 33 mM (high glucose) glucose. Postconfluency, the cells were exposed for 48 h to high glucose in the absence or presence of 16 microg/ml enoxaparin and tested for monocyte adhesion, expression of cell adhesion molecules, and translocation of nuclear transcription factor-kappaB (NF-kappaB), using adhesion assays, enzyme-linked immunosorbent assay (ELISA), and Western blotting. Statistical data revealed that treatment with enoxaparin resulted in a significant decrease in monocyte adhesion, expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and E-selectin, translocation of NF-kappaB, as well as in the level of intracellular reactive oxygen species. These results suggest that enoxaparin reduces the high glucose induced activation of endothelial cells by inhibiting monocyte adhesion through a mechanism that involves cell adhesion molecules and NF-kappaB. PMID- 14522368 TI - The role of chemotherapy in brain metastases. AB - Despite the widely held belief of the resistance to chemotherapy of brain metastases, central nervous system metastases of a malignancy are equally sensitive to chemotherapy as its metastases elsewhere in the body. This is due to the fact that the blood-brain barrier is disrupted in contrast enhancing brain metastases, and does not limit the response to chemotherapy. Therefore, the response rate of the primary tumour. Up-front chemotherapeutic treatment instead of radiotherapy of brain metastases should therefore be based on the chemosensitivity of the primary tumor to the used regimen, and not on the question whether the used agent penetrates an intact blood-brain barrier. First line chemotherapy for brain metastases or with only minor neurological signs and symptoms, and who have an indication for systematic chemotherapy for metastases elsewhere in the body. In contrast, central nervous system micrometastases may hide behind an intact barrier, and this may be clinically relevant in patients that can be cured with chemotherapy (like in small cell lung cancer). Cytochrome P450 3A4 inducing anti-epileptic drugs like phenytoin, carbamazepine and phenobarbital may significantly increase the metabolism of many chemotherapeutic agents like CPT11 and paclitaxel (but also of newer biological agents like many tyrosine kinase inhibitors). These anti-epileptic drugs should be avoided in patients requiring chemotherapy with agents metabolised through the cytochrome P450. PMID- 14522369 TI - Nasopharyngeal carcinomas: an update. AB - Among the group of head and neck cancers, nasopharyngeal carcinomas (NPC) represent a distinct entity in terms of their epidemiology, clinical presentation, biological markers, carcinogenic risk factors, prognostic factors, treatment and outcome. Undifferentiated NPC (UCNT), the most frequent histological type, is endemic in certain regions, especially in South East Asia. The disease has also been associated with the presence of the Epstein-Barr Virus (EBV). Although NPC is a radiosensitive and chemosensitive tumour, a substantial number of patients develop local recurrence or distant metastases. For patients with locoregional advanced disease, it is well known that conventional radiotherapy is insufficient in terms of both the local control rates and distant metastases. New techniques of radiation and new combined radiotherapy and chemotherapy modalities have been evaluated in numerous clinical trials in recent years. The purpose of this article is to review the current knowledge in terms of the epidemiology, biology, prognosis, management and outcome of patients with NPC. PMID- 14522370 TI - Why and how to study the fate of cancer survivors: observations from the clinic and the research laboratory. AB - This paper provides a rationale for the importance of studying health outcomes in cancer survivors, including their growing numbers, the need for detailed information on the short and long-term effects of treatment, and the interactions of cancer with other comorbid conditions. However, there are a number of challenges associated with this research. Identification and recruitment of cancer survivors is not always easy, and linking their current health status to details of medical treatment is important, but not always feasible. National surveys and databases are a potential source of information on survivors, but may not provide sufficient details for this type of research. Strategies need to be developed to plan for long-term outcome studies in cancer patients from the very beginning of treatment to obtain the most comprehensive data on the outcomes of survivors. PMID- 14522371 TI - Mismatch repair and response to DNA-damaging antitumour therapies. AB - Most antitumour therapies damage tumour cell DNA either directly or indirectly. DNA damage responses, and particularly DNA repair, influence the outcome of therapy. Because DNA repair normally excises lethal DNA lesions, it is intuitive that efficient repair will contribute to intrinsic drug resistance. Indeed, in certain circumstances reduced levels of DNA nucleotide excision repair are associated with a good therapeutic outlook (Curr Biol 9 (1999) 273). A paradoxical relationship between DNA mismatch repair (MMR) and drug sensitivity has been revealed by model studies in cell lines. This suggests that connections between MMR and tumour therapy might be more complex. Here, we briefly review how MMR deficiency can affect drug resistance and the extent to which loss of MMR is a prognostic factor in certain cancer therapies. We also consider how the inverse relationship between MMR activity and drug resistance might influence the development of treatment-related malignancies which are increasingly linked to MMR defects. PMID- 14522372 TI - The emergent role of focal liver ablation techniques in the treatment of primary and secondary liver tumours. AB - Only 20% of patients with primary or secondary liver tumours are suitable for resection because of extrahepatic disease or the anatomical distribution of their disease. These patients could be treated by ablation of the tumour, thus preserving functioning liver. This study presents a detailed review of established and experimental ablation procedures. The relative merits of each technique will be discussed and clinical data regarding the efficacy of the techniques evaluated. A literature search from 1966 to 2003 was undertaken using Medline, Pubmed and Web of Science databases. Keywords were Hepatocellular carcinoma, liver metastases, percutaneous ethanol injection, cryotherapy, microwave coagulation therapy, radiofrequency ablation, interstitial laser photocoagulation, focused high-intensity ultrasound, hot saline injection, electrolysis and acetic acid injection. Ablative techniques offer a promising therapeutic modality to treat unresectable tumours. Large-scale randomised controlled trials are required before widespread acceptance of these techniques can occur. PMID- 14522373 TI - Incidence of clinically significant lymphoedema as a complication following surgery for primary operable breast cancer. AB - There has recently been considerable interest for the need for specialist lymphoedema nurses to be appointed in the NHS. However, we had noticed in our cancer follow-up clinics that the incidence of lymphoedema appeared to be very low. Treatment for primary breast cancer (>5 cm) has been surgery and low axillary sampling (ANS). Radiotherapy (RT) or axillary clearance is subsequently performed in patients found to be node positive. The patients are followed-up in the primary breast cancer (PBC) clinic weekly. Follow-up is initially at 3-month intervals up to 2 years and then 1 yearly indefinitely. We conducted a two phased study in patients being followed up in our post cancer clinic in order to identify the incidence of LE in these patients. Phase 1 involved symptomatic patients identified at routine follow up in a 15-week period and the number of patients reporting arm swelling was recorded. The aim of this was to provide an estimate to power a phase 2 study (prospective questionnaire based). Phase 2 was conducted over a 13-week period. All patients attending the clinic were administered modified FACT B4, EQ-50 and Speilberger questionnaires. A total of 1242 patients were examined and lymphoedema found in 5 (0.04%). Of these 5, 3 had undergone axillary clearance, 1 ANS plus radiotherapy and only 1 had ANS alone. A policy of ANS, with prophylactic treatment for lymph node positivity either by surgery or RT alone, gives a very low rate of lymphoedema. PMID- 14522374 TI - Is physical function a more appropriate measure than volume excess in the assessment of breast cancer-related lymphoedema (BCRL)? AB - The aim of this study was to objectively measure impairment of arm function in women with breast cancer-related lymphoedema (BCRL), and investigate possible associations between this, arm volume excess, and psychological morbidity as measured by the Medical Outcomes Study 36-item short form (SF-36) questionnaire. A total of 48 patients were recruited. Manual dexterity was significantly impaired in the affected arm, independent of dominant or non-dominant arm involvement, but was not associated with arm volume excess. Psychological morbidity was significantly impaired in the domains of 'physical function' and 'bodily pain' when compared with population controls. Degree of impairment in the 'physical function' domain correlated with the absolute level of objectively tested manual dexterity. Impairment of manual dexterity appears to have a greater impact than arm volume excess on the overall psychological morbidity associated with BCRL, suggesting that greater emphasis should be placed upon arm function in the assessment, treatment targeting, and monitoring of patients with this condition. PMID- 14522375 TI - Role of imprint cytology in the intraoperative evaluation of sentinel lymph nodes for malignant melanoma. AB - Controversy exists over the utility of different methods for intra-operative sentinel lymph node (SLN) evaluation in patients with malignant melanoma (MM). The aim of this study was to evaluate the role of intra-operative imprint cytology (IC) in patients with MM. 215 SLNs from 99 patients with MM were examined by IC and results compared with the results of permanent sections. 24 patients had MM deposits in their SLNs and this was confirmed by histological examination. Intraoperative IC was positive in 11 of these patients (46% sensitivity). In addition, there were three false-positive IC diagnoses (79% positive predictive value); one of these was due to contamination during the sectioning of the SLN. The specificity and the negative predictive values of the IC were 96 and 85%, respectively. IC is a valuable method of intra-operative SLN evaluation which can spare approximately half of the patients with clinically occult regional metastases from a second surgical procedure. However, special care must be taken to avoid false-positive results due to contamination. PMID- 14522376 TI - Fatigue in long-term survivors of Hodgkin's lymphoma; a report from the German Hodgkin Lymphoma Study Group (GHSG). AB - Although treatment regimens for Hodgkin's lymphoma have become more sophisticated, little is known about the prevalence of fatigue in long-term survivors. Therefore, we investigated the fatigue status of long-term survivors of Hodgkin's lymphoma and a control group using a pre-validated questionnaire. In 1995/1996, we contacted 1981 patients, who were enrolled in the German Hodgkin Studies HD 1-6. All patients were treated according to the treatment protocols HD1-3 (1981-1988) and HD 4-6 (1988-1993). The patients with a current status of complete remission were asked to complete a quality-of-life (QoL) questionnaire (European Organisation for Research and Treatment of Cancer Quality of Life Core 30 (EORTC QLQ C-30)) and a fatigue questionnaire (Multidimensional Fatigue Inventory (MFI)). The results were compared with the data from 935 controls, matched for age, gender and living area. Eight-hundred and eighteen questionnaires from the patients were available for analysis. The median time between the end of treatment and completing the questionnaire is 5.2 years. Fatigue levels of patients with Hodgkin's lymphoma are high, even years after treatment. Fatigue dimensions are significantly influenced by several clinical and non-clinical factors. Fatigue levels of Hodgkin's lymphoma patients are significantly higher than those of the control group. Further investigations are warranted to explore the effectiveness of treatment strategies for fatigue. PMID- 14522377 TI - Interobserver variability between general and expert pathologists during the histopathological assessment of large-core needle and open biopsies of non palpable breast lesions. AB - The purpose of this study was to assess whether general pathologists are able to make as accurate and reproducible a diagnosis on large-core needle biopsies as on open breast biopsy specimens. A total of 688 patients underwent a stereotactic large-core (14G) needle biopsy and subsequent surgical excision of 718 non palpable breast lesions. Forty-two pathologists from 10 departments of pathology (generalists) made a diagnosis on both the needle and open biopsy specimens. Afterwards, three pathologists and two radiologists with extensive experience in breast pathology (experts) diagnosed all of the biopsy specimens. The general pathologists made a similar histological diagnosis as the experts in 632 (88%) of the needle biopsies and 649 (90%) of the open biopsy specimens. Accordingly, the interobserver agreement for the diagnosis of large-core needle biopsies between the general and experts pathologists was excellent (kappa 0.83) and not significantly different from the interobserver agreement for the diagnosis of open breast biopsies (kappa 0.86). However, many inconsistencies were observed in the category of borderline lesions: only 24% of the large-core needle biopsies and 43% of the open biopsies with an expert diagnosis of 'borderline' were diagnosed similarly by the general pathologists. Additionally, the risk of benign/malignant inconsistencies between general pathologists and experts was approximately 1 in 55 for both needle and open biopsies. PMID- 14522378 TI - Impact of locoregional treatment on the early-stage breast cancer patients: a retrospective analysis. AB - Although adequate locoregional treatment improves local and regional control in early-stage breast cancer, uncertainty still exists about the role of locoregional therapy with respect to survival. To study the impact of surgery and radiotherapy on locoregional control and survival, we combined the data of three European Organisation for Research and Treatment of Cancer (EORTC) Breast Cancer Group trials including early-stage breast cancer patients with long-term follow up. Risk ratios (RR) were estimated for locoregional recurrence and overall survival using Cox regression models. All analyses were adjusted for tumour size, nodal status, age, adjuvant radiotherapy, adjuvant chemotherapy and trial. The combined data-set consisted of 3648 patients. The median follow-up period was 11 years. 5.9% of the patients who underwent mastectomy and 10.8% of the patients who underwent breast-conserving therapy had a locoregional recurrence (P<0.0001). The risk of death after breast-conserving therapy was similar compared with mastectomy (RR 1.07, P=0.37). Adjuvant radiotherapy after mastectomy was associated with a lower risk for locoregional recurrence (RR 0.43, P<0.001) and death (RR 0.73, P=0.001). Patients with 1-3 positive nodes benefited the most from radiotherapy after mastectomy. Breast-conserving therapy was associated with an impaired locoregional control. However, breast-conserving therapy was not associated with a worse overall survival. Adjuvant radiotherapy in mastectomised patients was associated with both a significantly superior locoregional control and overall survival. The effect of adjuvant radiotherapy was most profound in patients who had 1-3 positive nodes. PMID- 14522379 TI - Familial hepatoblastoma and APC gene mutations: renewed call for molecular research. AB - Recent findings have increased our understanding of the molecular mechanisms involved in the pathogenesis of hepatoblastoma and their relationship to the molecular pathology of familial adenomatous polyposis (FAP). Here, we describe hepatoblastoma in siblings who share a gene mutation for FAP inherited from their father. This observation confirms the link between these diseases and has implications for future molecular research. We also raise the question; should other members of 'at-risk' families be screened following a new diagnosis of either hepatoblastoma or FAP? PMID- 14522380 TI - The Norwegian founder mutations in BRCA1: high penetrance confirmed in an incident cancer series and differences observed in the risk of ovarian cancer. AB - We aimed to describe the penetrances of the four Norwegian founder mutations in BRCA1 (816delGT, 1135insA, 1675delA and 3347delAG) with regard to breast and ovarian cancers in families ascertained through cancer family clinics or a consecutive series of women with breast or ovarian cancer. We have extended the families as far as possible and tested all family members that asked for genetic testing. Penetrance is based upon counting the mutation carriers. The series contains sufficient numbers of mutation carriers to minimise variation in the estimates due to a limited sample set. The penetrances for all four mutations were high, both with respect to breast and ovarian cancers. This is in accordance with other reports from cancer family clinics, but contrasts with reports from population-based series of mutation carriers. Risks of first cancer (breast or ovarian), breast cancer, and ovarian cancer at age 50 years were 43, 30 and 17%, respectively. Corresponding risks at age 70 years were 84, 58 and 58%. Risks for breast cancer before age 30 years and for ovarian cancer before 35 years were low. Penetrances with regard to ovarian cancer were different for the four mutations. The risk of ovarian cancer was doubled in carriers of the 1675delA mutation when compared with the 816delGT mutation (24 versus 12% at age 50 years, P=0.004). The mutations analysed are high penetrance alleles. No differences in penetrance between the series ascertained through the cancer family clinic or the series of consecutive cancer patients was observed. There are discrepancies between our findings and the low penetrances reported for other mutations in other populations. This may be due to methodological differences, but may reflect differences between mutations and/or modifying factors in different populations. PMID- 14522381 TI - Differences in colorectal cancer survival between European and US populations: the importance of sub-site and morphology. AB - A previous study has shown a lower survival for colorectal cancer in Europe than in the United States of America (USA). It is of interest to examine the extent to which anatomical location and morphological type influence this difference in colorectal cancer survival. We analysed survival for 151,244 European and 53,884 US patients diagnosed with colorectal cancer aged 15-99 years during the period of 1985-1989, obtained from 40 cancer registries that contribute to the EUROCARE study from 17 countries, and nine Surveillance, Epidemiology and End-Results (SEER) registries in the USA. Cases included in the analysis were first primary malignant tumours (ICD-O behaviour code 3 or higher). Relative survival was estimated to correct for competing causes of mortality. The Hakulinen-Tenkanen multiple regression approach was used to examine the prognostic impact of sub site and ICD-O histology codes. Relative excess risks (RERs) derived from this approach estimate the extent to which the hazard of death differs from that in a reference region after adjustment for mortality in the general population. In order to explore geographical variation, we defined three groups of European registries within which survival rates were known to be broadly similar. The proportion of cases with unspecified sub-site was higher in Europe than the USA (10% versus 2%), but sub-site distributions were broadly similar in the two populations. With the exception of appendix, 5-year survival was 13-22% higher in the USA than in Europe for each anatomical sub-site. The proportion of non microscopically-verified cases was higher in Europe than the USA (16 versus 3%). Adenocarcinomas arising in a polyp (ICD-O-2 8210, 8261, 8263) were more frequent in the USA than Europe (13 versus 2%). Five-year survival was higher in the USA than Europe for each morphological group, with the exception of non microscopically-verified cases. When age, gender and sub-site were considered, RERs ranged from 1.52 to 2.40 for the European populations (with the USA as a reference). After inclusion of morphology codes, the range of RERs fell to between 1.28 and 1.86, mainly because of the high frequency of adenocarcinoma in polyps in the USA. This analysis suggests that the large survival advantage for colorectal cancer patients in the USA can only marginally be explained by differences in the distribution of sub-site and morphology. The main explanatory difference is the proportion of adenocarcinoma in polyps. PMID- 14522382 TI - Suicides among cancer patients in Estonia: a population-based study. AB - The objective of this study was to determine the suicide risk among cancer patients in Estonia. This risk was examined in a cohort of 65,419 persons diagnosed with cancer in 1983-1998. Standardised mortality ratios (SMR) were calculated using the suicide rates of the population of Estonia as a reference. During 192,078 person-years of follow-up between 1983 and 2000, 197 suicides occurred in the cohort. An increased suicide risk was found for men (SMR=1.73; 95% Confidence Interval (CI) 1.45-2.01), but not for women (SMR=0.50; 95% CI 0.37 0.66). Men had the highest risk 90-179 days following their diagnosis (SMR=4.27; 95% CI 2.81-6.21). During this time interval, among men, the risk was more pronounced for cancers of the oesophagus (SMR=35.63; 95% CI 9.71-91.22) and pancreas (SMR=14.53; 95% CI 1.76-52.50). This study provides further evidence that cancer is a risk factor for suicide, at least in men. PMID- 14522383 TI - Heparanase-1 gene expression in normal, hyperplastic and neoplastic prostatic tissue. AB - Heparanase-1 (Hpa-1) has been implicated in tumour invasion and metastasis. In the present study, we evaluated the clinicopathological significance of Hpa-1 mRNA expression in prostate cancer and non-cancerous prostatic tissue by one-step polymerase chain reaction (PCR) of laser microdissected prostatic gland cells. In addition, cell type-specific expression of Hpa-1 mRNA in prostatic tissue was analysed by in situ hybridisation. Hpa-1 mRNA expression was found in 50% of normal and 40% of hyperplastic prostatic tissue. In situ hybridisation showed that Hpa-1 mRNA was strongly expressed in prostate gland cells. Of the 26 prostate carcinomas tested, 42% were positive for Hpa-1 mRNA. However, in non cancerous prostatic tissue, Hpa-1 mRNA was significantly more often expressed than in less differentiated or more invasive prostate cancers (P<0.05). In situ hybridisation revealed only focal Hpa-1 mRNA expression in the neoplastic gland cells. Hpa-1 mRNA expression in the tumours significantly correlated with tumour differentiation and tumour stage (P<0.05). Our data indicate that Hpa-1 gene expression may be lost during dedifferentiation of prostatic gland cells. PMID- 14522384 TI - Studies with cytotoxic agents suggest that apoptosis is not a major determinant of clonogenic death in neuroblastoma cells. AB - We have previously reported the selection of a radioresistant human neuroblastoma cell line, Clone F, from IMR32 cells. We have shown that clonogenic radioresistance in these cells is accompanied by a reduced level of radiation induced apoptosis [Cancer Res 55 (1995) 4915]. Here, we measured the response of these lines to several cytotoxic agents, in terms of clonogenicity and apoptosis. In the clonogenic assay, the radioresistant line was also resistant to cisplatin, melphalan and doxorubicin, but not to perillyl alcohol. However, all these agents produced less apoptosis in the Clone F cells, except cisplatin, which failed to induce any apoptosis in either cell line. Reduced apoptosis cannot be the cause of the Clone F cells' resistance to cisplatin. By extension, the Clone F cells' resistance to radiation and other agents cannot be due to diminished apoptosis either. Based on these results, apoptosis may not be a useful surrogate for clonogenic outcome. PMID- 14522386 TI - [Improvement in analgesia for ambulatory surgery]. PMID- 14522385 TI - Growth inhibition through activation of peroxisome proliferator-activated receptor gamma in human oesophageal squamous cell carcinoma. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) heterodimerises with retinoid X receptor alpha (RXRalpha) and is thought to be a novel therapeutic target for human malignancies. We evaluated the ability of troglitazone (TRO) alone or in combination with 9-cis retinoic acid (9CRA), ligands of PPARgamma and RXRalpha, respectively, to inhibit the growth of oesophageal squamous cell carcinoma (OSCC). All 10 tested OSCC cell lines of a KYSE series expressed PPARgamma and RXRalpha at both the mRNA and protein levels. In four tested cell lines, TRO inhibited growth, and a synergistic effect was observed with simultaneous 9CRA application. In KYSE 270 cells, a luciferase reporter assay showed that the simultaneous application of TRO and 9CRA to the cells increased the relative luciferase activity approximately 20-fold compared with the controls without TRO or 9CRA application. In this cell line, flow cytometry demonstrated that combined treatment with TRO and 9CRA greatly increased the sub-G1 phase, and Hoechst 33342/propidium iodide (PI) staining showed that apoptotic cell death was mainly induced through ligand treatment. In addition, implanted tumours in nude mice showed significant inhibition of tumour growth when treated with TRO. These results suggest that the PPARgamma/RXRalpha heterodimer may be a new therapeutic target for OSCC. PMID- 14522387 TI - [Impact of 2 strategies of analgesia on postoperative pain after emergency hand surgery performed on an ambulatory basis]. AB - OBJECTIVES: The assessment of the impact of various analgesic regimens on postoperative pain at home, following emergency and ambulatory hand surgery. STUDY DESIGN: Prospective comparative study of "before" and "after" groups by phone questionnaire on the first postoperative day. PATIENTS: All emergency and ambulatory surgery patients undergoing hand surgery in a specialist university hospital unit. METHODS: a) "Before" group (120 patients): analysis of current analgesic practice in the service (prescriptions by surgeon on duty), assessment of the incidence of pain scores with an intensity >/= 4 on a numerical score of 0 to 10, and identification of the most painful surgical interventions; b) formulation for these surgical interventions of multimodal analgesic protocols drown up by the anaesthetists; c) "After" group (51 patients): assessment of the efficacy of these analgesic protocols through comparison with the "before" group. RESULTS: a) "Before" group: the total incidence of pain scores with an intensity >/= 4 was 42% and in the case of conditions involving a joint or sepsis, 88%. Dextropropoxyfen-paracetamol alone was prescribed in 66% of cases and was associated with a 48% analgesic failure rate. Non-steroidal anti-inflammatory drugs were prescribed in 14% of cases, tramadol in 7% and a combination of drugs in 15%; b) "After" group: the incidence of pain scores with an intensity >/= 4 decreased to 21% in surgical interventions involving joints or sepsis (p < 0.001). CONCLUSION: Multimodal analgesic regimens are more effective than free prescriptions. This study underlines the need to further educate surgeons in this area. Despite these written protocols, there remains a 21% incidence of pain at home on day one. PMID- 14522388 TI - [Postoperative nausea and vomiting: cost and advantage induced by a quality insurance programme. Part I: post anaesthesia care unit]. AB - OBJECTIVES: To report the efficiency and direct cost induced by a quality insurance programme (QIP) aimed to reduce the incidence of post-operative nausea and vomiting (PONV) in the post-anaesthesia care unit. TYPE OF STUDY: Prospective and descriptive study. METHODS: The occurrence and treatment of PONV were prospectively recorded in the post-anaesthesia care unit in a standardised file before and following therapeutic recommendations made by volunteers component of a QIP. Prophylaxis was administered to patients according to a score of risks based on the type of surgery and patient characteristics. The real cost induced by this change of practice was evaluated according to pharmacists' accounts over 1 year. RESULTS: The QIP was associated to a 33.5% reduction of PONV relative risk and a 50% increase in prophylactic treatments. When this reduction of PONV was extrapolated to the total number of anaesthesia over 1 year, this QIP avoided PONV in 770 patients each year. The increase of cost reached 3572 as the cost by patient receiving a prophylactic treatment increased from 0.66 to 1. CONCLUSION: Despite an increase in the individual cost of PONV prophylaxis, the selection of patients by a "home" score of risk limits the indications of prophylactic treatment. Thus the increase in cost remains in acceptable ranges. PMID- 14522389 TI - [Prospective assessment of information programme for blood transfusion in critical care patients]. AB - OBJECTIVES: Assessment of oral and written information programme for blood transfusion in critical care patients and study of factors associated with the biological follow-up. STUDY DESIGN: Prospective study in one intensive care unit. PATIENTS AND METHODS: All blood recipients in 2000 were orally informed of transfusion process. In regard to the French haemovigilance regulation, written paper was given to the patient or passed on to the next hospital physician. Four months later, a questionnaire was proposed to the patients. RESULTS: In 2000, blood transfusion was performed for 102 patients and data were obtained for 59 patients (34 number of survivor patients, follow-up was impossible for eight and one refused the questionnaire). Quality assurance programme permit a global orally and written information of critical care patients. However, thirty-eight patients (64%) remembered oral information. Patients were younger (55 +/- 22 vs 68 +/- 13, OR = 1.04; p = 0.03). Biological follow-up was obtained for 22 patients (37%), particularly when written information was directly given to the patient (22/24 vs 2/35; OR = 335; p < 0.0001). CONCLUSION: Biological follow-up was obtained for only one third of blood recipients, because, in first, only 64% of critical care patients remembered oral information, particularly the younger patients. Perhaps, written information systematically given to the patient could permit a high biological follow-up. This study confirmed the lack of sensitivity of all hospital physician for haemovigilance. PMID- 14522390 TI - [Risk factors for amoxicillin-clavulanate-resistant Escherichia coli in ICU patients]. AB - OBJECTIVE: To determine risk factors of infections with amoxicillin-clavulanate resistant Escherichia coli in ICU patients. STUDY DESIGN: Prospective, consecutive sample survey study. PATIENTS: A consecutive series of 133 patients from whom culture results were positive for E. coli during their ICU stay. METHODS: Risk factors analysed included demographics, comorbid conditions, and antimicrobial drug exposure. Univariate and multivariate analysis were performed. RESULTS: Multivariate logistic regression analysis identified only one significant independent factor associated with the emergence of amoxicillin clavulanate-resistant E. coli: prior use of amoxicillin (odds ratio: 5.45). CONCLUSION: Clinicians should avoid administering amoxicillin-clavulanate as empiric therapy for possible E. coli infection in patients that have recently been treated with amoxicillin. PMID- 14522391 TI - [Starting experience with non-invasive ventilation in paediatric intensive care unit]. AB - Non-invasive ventilation in pressure support (NIV) is well described in the adult and child over 5 years. However, its use in children less than 1 year of age remains anecdotal. We report our preliminary experience with the use of NIV in six children aged from 5 days to 10 months. NIV was delivered with a flow generator (VPAP IIST, Resmed Ltd, North Ryde, NSW, Australia) in association with specific tubings and a nasal mask. The use of NIV resulted in a significant decrease of both the respiratory rate (from 53 to 39 breaths per min, p < 0.01) and the PvCO(2) (from 9.33 to 6.28 kPa, p < 0.01). These results show that NIV can be used in children under 1 year of age with improvement of physiological parameters. PMID- 14522392 TI - [Activation of adhesion molecules in patients with septic shock]. AB - OBJECTIVE: To analyze the pattern of adhesion molecules in patients with septic shock. Data sources. - References obtained from Pubmed databank. DATA EXTRACTION: Models of inflammation linking endothelial dysfunction, adhesion molecules and septic states were analyzed. DATA SYNTHESIS: The endothelium has been identified as the central effector in the inflammatory response. Adhesion molecules are strongly involved in the inflammatory process by modulating the leukocyte trafficking. The most important adhesion molecules are the selectins (E-, L-, and P-selectins) and members of the immunoglobulin superfamily (intercellular adhesion molecule-1 and vascular cell adhesion molecule-1). Plasma levels of these molecules are increased in septic shock patients, which may be related to a marked activation of the endothelium. However, a dichotomous profile is observed between plasma and tissue expression. The inhibition of adhesive molecule actions could make it possible to control the inflammatory response. PMID- 14522394 TI - [Non cardiac surgery in patient with coronary stenting: think sirolimus now!]. AB - We describe a severe preoperative cardiogenic shock in a patient scheduled for a breast surgery. The cardiogenic shock was in relation with thrombosis of two sirolimus-eluting stents received 3 months ago. A percutaneous transluminal coronary angioplasty was successfully performed. The patient recovered well after a 1-day treatment including intraaortic balloon counter pulsation and dobutamine infusion. We discuss about the ideal timing to plan surgery and how to manage the shift of antiplatelet agents. PMID- 14522393 TI - [Postoperative pancreatitis after non abdominal surgery]. AB - A 35-year-old man developed two episodes of postoperative pancreatitis. The first one occurred after knee surgery and the second one 6 years after surgery for excision of haemorrhoids. Induction of anaesthesia associated propofol 200 mg and sufentanil 15 microg. Anaesthesia was maintained with sevoflurane administered through a laryngeal mask. The possible association between the drugs used during anaesthesia and the postoperative pancreatitis is discussed. PMID- 14522395 TI - [Spleen trauma during pregnancy (a case report)]. AB - Splenic injury during pregnancy after blunt abdominal trauma is rare. The physiological and anatomical changes during pregnancy modify the clinical, biological and radiological signs. The authors report a case of a 30-year-old woman in the 32nd week of pregnancy, with an isolate splenic injury after a motor vehicle accident. Splenectomy was realised 4 days after an initial decision of abstaining from surgery. In pregnancy, the frequency of recurrent bleeding, the possibility of concomitant uterine injuries and the risk of foetal loss should modify the surgeon's attitude to splenic rupture after abdominal blunt trauma. PMID- 14522396 TI - [Fistulas diagnosis by amylases levels in drainage fluids after gastrectomy]. PMID- 14522397 TI - [Caspofungin: indication in Candida albicans-induced severe sepsis?]. PMID- 14522398 TI - [Upper digestive endoscopy in children: is laryngeal mask useful and safe?]. PMID- 14522399 TI - [Proposal of a score to predict difficult neuroaxial block]. PMID- 14522400 TI - [Proceedings of a congress of the Sfar 2002]. PMID- 14522401 TI - Infection and inflammation in the cardiovascular system. PMID- 14522402 TI - Myocarditis and dilated cardiomyopathy: an inflammatory link. AB - Myocarditis is a complex disease because multiple pathogenetic mechanisms play a role. While these mechanisms appear to act in a chronological cascade, they undoubtedly overlap in some cases, rendering diagnosis and treatment difficult. Ultimately, dilated cardiomyopathy (DCM) may result. A multitude of still circumstantial evidence points to a major role of viral myocarditis in the etiology of DCM. The common presence of viral genetic material and viral proteins in the myocardium of patients with DCM provides the most compelling evidence, but proof of causality is still lacking. Nevertheless, because of the striking increase in heart failure prevalence in recent years, anti-viral and anti inflammatory therapies should be developed for their potential to prevent or ameliorate DCM. PMID- 14522403 TI - Myocarditis and inflammatory cardiomyopathy: microbiological and molecular biological aspects. AB - Myocarditis is an inflammatory disease of the myocardium associated with cardiac dysfunction. The natural history of myocarditis is frequently characterised by the evolution in dilated cardiomyopathy. Due to its variable clinical manifestation from latent to very severe clinical forms, such as acute congestive heart failure and sudden death, its prevalence is still unknown and probably underestimated. In spite of the development of various diagnostic modalities, early and definite diagnosis of myocarditis still depends on the detection of inflammatory infiltrates in endomyocardial biopsy specimens according to the Dallas criteria. Routine application of immunohistochemistry, used for identification and characterisation of inflammatory cell populations, has now significantly increased the sensistivity of the diagnosis of inflammatory cardiomyopathy. Various molecular techniques, such as PCR, gene sequencing and real-time PCR, often applied on the same endomyocardial specimen, have become an essential part of the diagnostic armamentarium for rapid, specific and sensitive identification of infective agents. The correct application of molecular techniques will allow increasingly more information to be obtained: new epidemiology, new patient risk stratification and overall more appropriate medical treatment. PMID- 14522404 TI - Infection and inflammation and the coagulation system. AB - Severe infection and inflammation almost invariably lead to hemostatic abnormalities, ranging from insignificant laboratory changes to severe disseminated intravascular coagulation (DIC). Systemic inflammation results in activation of coagulation, due to tissue factor-mediated thrombin generation, downregulation of physiological anticoagulant mechanisms, and inhibition of fibrinolysis. Pro-inflammatory cytokines play a central role in the differential effects on the coagulation and fibrinolysis pathways. Vice-versa, activation of the coagulation system may importantly affect inflammatory responses by direct and indirect mechanisms. Apart from the general coagulation response to inflammation associated with severe infection, specific infections may cause distinct features, such as hemorrhagic fever or thrombotic microangiopathy. The relevance of the cross-talk between inflammation and coagulation is underlined by the promising results in the treatment of severe systemic infection with modulators of coagulation and inflammation. PMID- 14522405 TI - Infections and endothelial cells. AB - Systemic infection by various pathogens interacts with the endothelium and may result in altered coagulation, vasculitis and atherosclerosis. Endothelium plays a role in the initiation and regulation of both coagulation and fibrinolysis. Exposure of endothelial cells may lead to rapid activation of coagulation via tissue factor (TF) expression and the loss of anticoagulant properties by impairment of antithrombin III, TF pathway inhibitor (TFPI) and the protein C system. Endothelial-derived plasminogen activator inhibitor (PAI) is essential for the regulation of fibrinolysis and impaired endothelial function leads to imbalance in fibrinolysis, resulting in a procoagulant state. The interaction between inflammation and coagulation, soluble adhesion molecules and circulation endothelial cells is important in the pathogenesis of an unbalanced haemostatic system. Rather than being a unidirectional relationship, the interaction between inflammation and coagulation appears to be significant. In the crosstalk, the endothelium is playing a pivotal role. PMID- 14522406 TI - Molecular basis of endothelial dysfunction in sepsis. AB - Sepsis is one of the major causes of mortality in critically ill patients and develops as a result of the host response to infection. A complex network of events is set into motion in the body by the infection and results in the pathogenesis of sepsis. This review article focuses on the molecular mechanisms and components involved in the pathogenesis of sepsis with a major emphasis on the endothelium. This includes sepsis-inducing bacterial components (e.g. endotoxins), cellular targets of these molecules and their responses, host reactions, intracellular and cytokine networks, individual susceptibility and new therapeutic targets in sepsis treatment. PMID- 14522407 TI - Toll-like receptors in cardiovascular diseases. AB - The Toll-like receptor family recognizes mostly exogenous ligands like bacterial lipopolysaccharides or DNA and activates the inflammatory cell. Evidence is accumulating that these Toll-like receptors are important in cardiovascular pathologies. Recently, expression of Toll-like receptors in arterial and myocardial cells has been shown and mouse knockout and human studies on polymorphisms point to a role of Toll-like receptor 4 in neointima formation and atherosclerosis. It is now becoming clear that these receptors not only serve as receptors for pathogen-associated molecular patterns but are also involved in the initiation and progression of cardiovascular pathologies. PMID- 14522408 TI - Type II secretory phospholipase A2 in cardiovascular disease: a mediator in atherosclerosis and ischemic damage to cardiomyocytes? AB - Inflammatory reactions contribute to the pathogenesis of cardiovascular conditions such as atherosclerosis and ischemic damage in acute myocardial infarction (AMI). Among the mediators involved in inflammation are secretory phospholipase A2 group II (sPLA2-II) enzymes. Though some cells constitutively express sPLA2-II, the synthesis by cells such as hepatocytes is typical for an acute-phase reactant. Recent literature suggests multiple roles for sPLA2-II in cardiovascular disease. In this review we discuss the role of sPLA2-II in various in vivo and in vitro models of atherosclerosis or AMI, including the therapeutic perspective of sPLA2-II inhibitors. It was concluded that sPLA2-II appears to be an important inflammatory mediator of cardiovascular disease. PMID- 14522409 TI - T lymphocytes in atherogenesis-functional aspects and antigenic repertoire. PMID- 14522410 TI - HIV infection, highly active antiretroviral therapy and the cardiovascular system. AB - Cardiovascular complications in the course of human immunodeficiency virus (HIV) infection are multifactorial and may be caused by the virus itself or by the related opportunistic infections and neoplasms. Highly active antiretroviral therapy (HAART) has prolonged many patients' lives, but many cardiac sequelae of HIV are not affected by HAART and continue to develop even with treatment. In addition, HAART itself causes in a high proportion of patients a metabolic syndrome, characterized by lipodystrophy/lipoatrophy, dyslipidemia and insulin resistance that may be associated with an increase in peripheral artery and coronary artery diseases. Careful cardiovascular evaluation in the course of HIV disease can identify cardiac complications early enough to treat. All HIV infected patients candidate to antiretroviral therapy and patients already under treatment should undergo an assessment that includes the evaluation of the cardiovascular risk with the available guidelines. PMID- 14522411 TI - Pathophysiology of the heart in Chagas' disease: current status and new developments. AB - In the present review we have summarized remarkable historical data on Chagas' disease studies putting special emphasis on histopathological findings and pathogenetic theories as well as recent discoveries based on the use of advanced modern technologies in pathology and immunology. A unified theory that links almost all of these findings is proposed. Chronic cardiac Chagas' disease represents the result of a close interaction between the host and the parasite, causing different clinical pictures: patients with an efficient immune response may adequately circumvent the parasitic infection and the individual will develop the indeterminate form. Deficient immune response of the host and/or a high initial parasitemia favor an immune imbalance that might lead to development of a permanent inadequate immunological response against the parasite. The inflammatory response, which is probably recurrent, undergoing periods of more accentuated exacerbation, is most likely responsible for progressive neuronal damage, microcirculatory alterations, heart matrix deformations and consequent organ failure. PMID- 14522412 TI - Cardiomyopathy in a murine model of AIDS: evidence of reactive nitrogen species and corroboration in human HIV/AIDS cardiac tissues. AB - OBJECTIVE: Cardiomyopathy and other vascular complications are now recognized as significant components of HIV/AIDS pathogenesis. Although the mechanisms involved in cardiomyopathy are poorly defined, a role for direct retroviral action and/or focal infiltration of activated immune cells have been postulated. Here we investigated mechanisms in retrovirus associated cardiomyopathy using a well defined mouse model of acquired immunodeficiency. METHODS: Mice were dosed with LPBM5 retrovirus; cardiac performance was assessed by echocardiography followed by tissue collection at 5 and 10 weeks post-infection. RESULTS: Contractile deficits were observed at 5 and 10 weeks post-retrovirus infection and preceded the development of overt immunodeficiency. Selective and widespread cardiac infiltration of CD68+ cells, but not neutrophils, mast cells, or eosinophils was also observed at both 5 and 10 weeks. LPBM5 retrovirus was readily detectable in cardiac samples by RT-PCR. Time dependent increases in cardiac protein nitration (biomarker of reactive nitrogen species) were observed and were correlated to the extent of cardiac dysfunction whereas no changes in NOSII occurred at 5 and 10 weeks. We corroborated the mouse findings using cardiac tissues and clinical findings from human HIV/AIDS autopsies. CONCLUSIONS: These studies demonstrated that cardiac myocyte protein nitration in AIDS related cardiomyopathies, rather than focal immune cell lesions characterize retrovirus associated cardiomyopathies and differentiate them from non-retroviral cardiomyopathies. PMID- 14522413 TI - Impairment of glucose metabolism in hearts from rats treated with endotoxin. AB - OBJECTIVE: In patients and animals with sepsis or critical illness, the mechanical function of the heart is often impaired. Although these conditions are accompanied by dramatic metabolic and hormonal changes, little is known about alterations of cardiac metabolism. In this study, we assessed the impact of an endotoxin-induced inflammation on cardiac glucose utilization. METHODS: Bacterial endotoxin (1 mg/kg lipopolysaccharide from Salmonella typhimurium, LPS) was injected intravenously to rats. Six hours after LPS application, hearts were isolated and perfused in the Langendorff mode. RESULTS: Left ventricular pressure was reduced by 50% in hearts from LPS-treated rats, compared to those from saline injected control animals. With glucose as the sole fuel, there was no difference in glycolysis between the groups. However, on addition of beta-hydroxybutyrate (an alternative fuel which inhibits phosphofructokinase via an increased citrate level), the glycolytic rate in the LPS group was 44 and 48% lower (in basal, and insulin-stimulated conditions, respectively; P<0.01) than in control hearts. At the end of perfusions with beta-hydroxybutyrate and insulin, the cardiac citrate content was 40% higher in LPS vs. controls (P<0.001). In addition to the reduced glycolysis, the insulin-dependent increase of cardiac glycogen was 77% smaller in LPS hearts. The difference between LPS and control glycolysis was abolished if the hearts were perfused with the ceramidase inhibitor N-oleyl-ethanolamine (5 microM), and also with the cyclooxygenase-2 inhibitor NS-398 (10 microM), or the thromboxane A2 receptor antagonist SQ-29548 (1 microM). CONCLUSION: The inflammatory reaction caused by endotoxin impairs cardiac glucose metabolism (and in particular, the action of insulin) in at least two ways: through the exacerbation of the counterregulatory effect of alternative fuels on glycolysis, and through a reduction in net glycogen synthesis. Impairment of glycolysis may be mediated by a sphingomyelin derivative, and COX-2-derived thromboxane A2. PMID- 14522415 TI - Preprocedural serum levels of acute-phase reactants and prognosis after percutaneous coronary intervention. AB - OBJECTIVE: In this study we evaluate the value of baseline concentrations of acute-phase reactants on prognosis after percutaneous coronary intervention (PCI). METHODS: Blood samples were drawn immediately before PCI to measure baseline concentrations of C-reactive protein (CRP), interleukin-6 (IL-6), lipoprotein(a) (Lp(a)), and fibrinogen. Follow-up data were collected at 8 months. Repeat PCI, CABG, myocardial infarction, and death were recorded as major adverse clinical events (MACE). Furthermore the recurrence of angina pectoris was noted. RESULTS: The study included 600 consecutive patients after a successful PCI. Sixty-four percent of the patients were stented. The mean age was 61.6 years and 68.9% were male. CRP levels were significantly higher in patients who were to have repeat angina as compared with those who were not (P=0.0322). IL-6 levels were not correlated with angina or MACE. Lp(a) and fibrinogen concentrations were both significantly related to MACE (P=0.0337 and P=0.0253, respectively). CONCLUSION: Our study clearly supports the role of inflammation in restenosis after PCI as measured in statistically higher levels of Lp(a) and fibrinogen in patients with MACE and CRP in patients with repeat angina. PMID- 14522414 TI - Effects of enalapril on disseminated intravascular thrombin formation during systemic inflammation. AB - BACKGROUND: Tissue factor (TF), the main trigger of coagulation is important in the propagation of cardiovascular diseases. Based on an in vitro study, we hypothesised that enalapril may blunt the endotoxin-induced, TF-triggered coagulation in humans. METHODS: In a randomised, controlled trial, 30 healthy male volunteers received 2 ng/kg of lipopolysaccharide (LPS) after pre-treatment with placebo or enalapril for 5 days or with enalapril 2 h before LPS infusion. RESULTS: Infusion of LPS increased interleukin-6 levels 400 fold, and induced a 10-fold increase in prothrombin fragment, a fourfold increase in D-dimer, and a fivefold increase in plasmin-antiplasmin complexes. However, pre-treatment with enalapril did not blunt LPS-induced coagulation. CONCLUSIONS: Our trial provides evidence against a modulatory role of angiotensin converting enzyme in LPS induced, TF-triggered coagulation. PMID- 14522416 TI - Enhanced expression of inflammatory cytokines and activation markers in T-cells from patients with chronic heart failure. AB - OBJECTIVE: Increasing evidence supports a role for inflammation in chronic heart failure (CHF). However, the source and the mechanism for this immune activation are unknown. To address this issue we investigated the gene expression of cytokines and the surface expression of activity markers in T-cells and monocytes from CHF patients and healthy controls. METHODS: Gene expression of cytokines was analysed by real-time RT-PCR and activation markers by flow cytometry in 14 CHF patients and nine healthy controls. Surface expression of activation markers for T-cells and monocytes were analysed by flow cytometry. RESULTS: T-cells from CHF patients showed enhanced gene expression of chemokines, ligands in the tumor necrosis factor superfamily, as well as the inflammatory cytokines interferon gamma and interleukin-18 with similar pattern in ischemic (n=5) and idiopathic cardiomyopathy (n=9). In contrast, no differences in cytokine gene expression were found comparing monocytes from CHF patients and controls. Moreover, T-cells from CHF patients had enhanced surface expression of the activation markers CD69 and CD25, while there was no upregulation of the monocyte activation marker CD32 in these patients. CONCLUSION: T-cells may be a part of the inflammatory response during CHF independent of the etiology of the disorder. Intervention preventing unwanted T-cell activation could represent a new target in the treatment of CHF. PMID- 14522417 TI - Zidovudine-induced alterations in the heart and vascular smooth muscle of the rat. AB - OBJECTIVE: We investigated the effects of zidovudine (AZT) on cardiac and vascular smooth muscle function and morphology in rats. METHODS: Four adult male Wistar-Kyoto rats received AZT in drinking water for 240 days; four rats served as controls. Echocardiographic examination and systolic blood pressure (SBP) measurement were performed. At the end of treatment the rats were sacrificed, their hearts were weighed and vascular smooth muscle contractile and relaxing properties were evaluated in vitro on endothelium-intact aortic rings. Morphological studies were performed on cardiac and aortic myocytes by light and electron microscopy. RESULTS: AZT-treated rats (AZT-Rs) showed higher SBP, greater heart weight and, as revealed by echocardiography, greater interventricular septum thickness. Electron microscopy revealed mitochondrial swelling in myocardiocytes in AZT-Rs. Reduced response to contractile stimuli and enhanced relaxation in response to charbacol were observed in the aortic rings of AZT-Rs. The aortic myocytes of AZT-Rs contained apparently unaffected ultrastructural features, but light microscopy suggested their marked enlargement. CONCLUSIONS: AZT treatment for 240 days in rats induces a modest increase in SBP, hypertrophy of the interventricular septum and modified vascular smooth muscle responsiveness. The role of mitochondria in these AZT-induced cardiovascular alterations remains to be established. PMID- 14522418 TI - Intra-amniotic lipopolysaccharide leads to fetal cardiac dysfunction. A mouse model for fetal inflammatory response. AB - OBJECTIVE: Intrauterine infection is associated with increased lipopolysaccharide (LPS) and proinflammatory cytokines in amniotic fluid. We hypothesized that intra amniotic LPS launches a fetal inflammatory response leading to cardiac dysfunction. METHODS: A mouse model was established. At 15-16 days of gestation, 52 fetuses of nine dams received LPS and 46 fetuses of nine dams vehicle intra amniotically. Five dams underwent a sham operation. Echocardiography was performed before and 6 h after the injection to obtain inflow and outflow blood velocity waveforms. Outflow mean velocity (V(mean)) and the proportions of isovolumetric relaxation (IRT%) and contraction (ICT%) times of the cardiac cycle were calculated. Pulsatility indices (PI) were calculated from the umbilical and intracranial arteries and the descending aorta. Pulsatility indices for veins (PIV) were obtained from ductus venosus. Toll-like receptor-4 (TLR4) and several other inflammatory mediators were determined using ELISA, immunohistochemistry, or ribonuclease protection assay. RESULTS: In the LPS group, outflow V(mean) was significantly lower, and ICT% and IRT% longer than in the other groups. LPS increased PIs, except in the intracranial arteries, which showed a decrease in PIs. In ductus venosus, PIVs were increased after LPS. LPS increased interleukin (IL)-6 in amniotic fluid and induced the expression of proinflammatory cytokines in placenta and fetal membranes, but not in lung. In fetal myocardium, TLR4 was constitutional. LPS induced the expression of IL-1beta and tumor necrosis factor (TNF)-alpha mRNA in myocardium, whereas inducible nitric oxide synthase (NOS2) protein and nitrotyrosine remained undetectable. CONCLUSIONS: As a response to endotoxin in amniotic fluid, fetal myocardium acutely generates cytokines and severe fetal cardiovascular compromise develops. These two may be linked through a mechanism that does not include NO. PMID- 14522419 TI - Placental infection with Chlamydia pneumoniae and intrauterine growth restriction. AB - BACKGROUND: The concept that low birth weight infants are more predisposed to coronary artery disease (CAD) in adulthood has been studied extensively. Although many infectious agents have been associated with intrauterine growth restriction (IUGR), Chlamydia pneumoniae an organism implicated in CAD has not been investigated. It was our aim to assess whether C. pneumoniae DNA is present in placental tissue and whether its detection is associated with IUGR. METHODS: Fifty-nine pregnant women were studied: 32 women had an uncomplicated pregnancy with no antenatal or post-natal evidence of IUGR. Twenty-seven women had pregnancies with ultrasonographically demonstrated IUGR, defined as foetal abdominal circumference measuring less than 2 S.D.s from the mean for gestational age. At the time of delivery, maternal blood and placental tissue samples were obtained. Placental samples were taken from four sites centrally and peripherally on the maternal and foetal side of the placentas and tested by nested polymerase chain reaction for C. pneumoniae DNA. IgG antibodies to C. pneumoniae were measured using microimmunfluorescence. RESULTS: C. pneumoniae DNA was detected in 44% of the placental tissue but there was no difference in the prevalence of bacterial DNA between the control and the low birth weight group (P=0.58). Additionally C. pneumoniae seropositivity did not differ between the index and control groups (78 vs. 70%, P=0.44). CONCLUSIONS: C. pneumoniae is present in placental tissue. Its presence however does not correlate with IUGR. Similarly, maternal C. pneumoniae seropositivity is not related to low birth weight. Thus C. pneumoniae infection is unlikely to play a role in the pathogenesis of IUGR. PMID- 14522420 TI - Nitric oxide synthase plays a role in Chlamydia pneumoniae-induced atherosclerosis. AB - OBJECTIVE: Chlamydia pneumoniae infection has been associated with atherosclerosis, although the mechanisms by which C. pneumoniae contribute to atherogenesis remain unclear. Altered production of nitric oxide, a known bactericidal and anti-inflammatory agent, represents one possible mechanistic link. To examine this issue, a diet-induced, hyperlipidemic mouse model of early atherosclerosis was used. METHODS: A series of intranasal inoculations of C. pneumoniae strain AR-39 were administered to mice lacking endothelial or inducible nitric oxide synthase and to normal controls. After 18 weeks on an atherogenic diet, atherosclerotic lesion area in the aortic sinus was measured using computer-assisted morphometry. RESULTS: In the absence of C. pneumoniae infection, diet-fed eNOS(-/-) mice developed enlarged fatty streak lesions of borderline significance in comparison to uninfected, wild-type mice, while the lesion area in uninfected, diet-fed iNOS(-/-) mice did not differ significantly from lesion area in wild-type animals. In contrast, lesion area in infected eNOS( /-) mice increased slightly, but not significantly in comparison to uninfected eNOS(-/-) mice. Lesion area in the infected iNOS(-/-) mice was significantly enlarged when compared to both uninfected iNOS(-/-) mice as well as to infected wild-type mice. CONCLUSIONS: These data suggest that production of nitric oxide by eNOS protects against development of fatty streak lesions in uninfected hyperlipidemic mice, but does not offer additional protection in infected hyperlipidemic mice, while iNOS may play a protective role, thus limiting chlamydial exacerbation of fatty streak lesions. PMID- 14522421 TI - Dendritic cells in the arterial wall express C1q: potential significance in atherogenesis. AB - OBJECTIVE: Dendritic cells (DCs) accumulate in atherosclerotic lesions but their characteristics and their role in atherogenesis are poorly understood. C1q, an element of the first component of complement, is expressed by interdigitating dendritic cells and follicular dendritic cells in the spleen. It has been suggested that C1q is involved in capturing immune complexes in the lymphoid tissue. Immune complexes are also detected in atherosclerotic lesions. The present study investigated whether C1q is expressed by DCs in the arterial wall. Because DCs accumulating within atherosclerotic lesions might originate from monocytes that infiltrate the intima from very early stages of atherosclerosis, C1q expression was also examined in monocyte-derived DCs in vitro. METHODS: Specimens of the aorta, carotid, mammary, popliteal and tibial arteries were obtained during operation. Expression of C1q in the arterial wall was studied by immunohistochemistry. The nature of cells expressing C1q was studied in sections double stained with antibodies to C1q and cell type specific markers including CD1a and S-100 (for identification of DCs), CD68 (macrophages), CD3 (T-cells), von Willebrand factor (endothelial cells), and smooth muscle alpha-actin (smooth muscle cells). In vitro, DCs were differentiated from human peripheral blood monocytes using GM-CSF and IL-4. Peripheral blood monocytes were differentiated to macrophages using M-CSF. The expression of C1q in monocytes and in vitro monocyte-derived DCs and macrophages was determined by RT-PCR, Western blotting, immunofluorescence microscopy and flow cytometry. RESULTS: In all the arterial specimens studied, DCs expressing C1q were detected. C1q was also found in macrophages, macrophage foam cells and in neovascular endothelial cells in atherosclerotic lesions, but no C1q expression was detected in T-cells and smooth muscle cells. In vitro analysis demonstrated that monocyte-derived DCs and macrophages express C1q but no C1q was detected in monocytes. CONCLUSION: C1q is expressed by DCs residing in the arterial wall as well as by monocyte-derived DCs in vitro. Expression of C1q occurs during differentiation of monocytes to DCs and macrophages and might be important in binding and trapping immune complexes in atherosclerotic lesions. PMID- 14522422 TI - Reactive oxygen species mediate cyclooxygenase-2 induction during monocyte to macrophage differentiation: critical role of NADPH oxidase. AB - OBJECTIVE: The objective of this study was to explore the relationship between monocyte differentiation into macrophages and cyclooxygenase-2 (Cox-2) expression, based upon the observation that high amounts of this enzyme, colocalizing mainly with macrophages, have been found in human atherosclerotic lesions. Moreover, the hypothesis that reactive oxygen species (ROS) could be important as mediators of Cox-2 expression during monocyte differentiation was verified. Although ROS are known as modulators of gene expression profile, their involvement in monocyte differentiation has not been explored previously. METHODS: Human adherent monocytes and the promonocytic cell line U937 were differentiated into macrophages by phorbol ester (PMA). Cox-2 was evaluated in terms of protein, mRNA and activity. Intracellular ROS formation was measured by the oxidant sensitive dye 2',7'-dichlorofluorescein diacetate. NADPH oxidase subunit p47(phox) was evaluated by Western blot analysis. RESULTS: Functionally active Cox-2 is expressed during PMA-induced monocyte transition into macrophages and ROS driven by the NADPH oxidase play a critical role in this event. CONCLUSION: Monocyte differentiation into macrophages, possibly triggered by unquenched ROS, may contribute to the increased inflammatory response within atheromata. PMID- 14522424 TI - TIMP-2 and PAI-1 mRNA levels are lower in aneurysmal as compared to athero occlusive abdominal aortas. AB - OBJECTIVE: Significant alterations of the vascular wall occurs in abdominal aortic aneurysm (AAA) and atherosclerotic occlusive disease (AOD) that ultimately may lead to either vascular rupture or obstruction. These modifications have been ascribed to one or a group of proteases, their inhibitors or to the matrix macromolecules involved in the repair process without considering the extent of the observed variations. METHODS: The mRNA steady-state level of a large spectrum of proteolytic enzymes (matrix metalloproteinases: MMP-1, -2, -3, -8, -9, -11, 12, -13, -14; urokinase plasminogen activator: u-PA), their physiological inhibitors (tissue inhibitors of MMPs: TIMP-1, -2, -3; plasminogen activator inhibitor: PAI-1) and that of structural matrix proteins (collagens type I and III, decorin, elastin, fibrillins 1 and 2) was determined by RT-PCR made quantitative by using a synthetic RNA as internal standard in each reaction mixture. The profile of expression was evaluated in AAA (n=7) and AOD (n=5) and compared to non-diseased abdominal (CAA, n=7) and thoracic aorta (CTA, n=5). RESULTS: The MMPs -8, -9, -12 and -13 mostly associated with inflammatory cells were not or barely detected in CAA and CTA while they were largely and similarly expressed in AAA and AOD. Expression of protease inhibitors or structural proteins were only slightly increased in both pathological conditions with the exception of elastin which was reduced. The main significant difference between AAA and AOD was a lower expression of TIMP-2 and PAI-1 in the aneurysmal lesions. CONCLUSIONS: The remodeling of the aortic wall in AAA and AOD involves gene activation of a large and similar spectrum of proteolytic enzymes while the expression of two physiological inhibitors, TIMP-2 and PAI-1, is significantly lower in AAA compared to AOD. The repair process in the aneurysmal disease seems similar to that of the occlusive disease. PMID- 14522423 TI - Chronic inhibition of cyclooxygenase-2 does not alter plaque composition in a mouse model of advanced unstable atherosclerosis. AB - OBJECTIVE: Inflammation contributes to atherosclerotic plaque initiation and progression. Recent studies suggest that anti-inflammatory drugs such as cyclooxygenase-2 (Cox-2) inhibitors have anti-atherogenic effects. The current study was designed to investigate whether administration of a Cox-2 inhibitor to older apolipoprotein E deficient (apo E-/-) mice with established lesions alters the composition and increases the stability of the lesions. METHODS AND RESULTS: The Cox-2 inhibitor Celecoxib was administered in chow to 26-week-old, male, apo E-/- mice exhibiting advanced, unstable atherosclerotic lesions within the innominate/brachiocephalic artery. Mice administered Celecoxib had no significant changes in serum cholesterol or the average cross sectional area of atherosclerotic lesion in the innominate artery after 15 weeks of treatment in comparison to non-treated control mice. Histological analyses of sections of the innominate artery demonstrated no significant changes in the frequency of markers of advanced and unstable atherosclerotic plaques, including intra-plaque hemorrhage, vascular calcification, thinning of the fibrous cap, size of the necrotic core and macrophage content. There were also no significant differences in the content of Cox-2 within the lesions. Quantitative real time polymerase chain reaction with mRNA isolated from the aorta of each mouse revealed no significant changes in the expression of tissue factor and inducible nitric oxide synthase. However, mRNA levels for MCP-1 were increased fivefold following 15 weeks of treatment with Celecoxib in comparison to non-treated control mice. CONCLUSIONS: These data suggest that Celecoxib has no effect on the composition of advanced atherosclerotic lesions in older apo E-/- mice. PMID- 14522426 TI - The African cherry (Prunus africana): can lessons be learned from an over exploited medicinal tree? AB - For the last 35 years, the African cherry (Prunus africana (Hook. f.) Kalm.) has been used in the treatment of benign prostatic hyperplasia and other disorders. The bark, from which the treatment is derived, is entirely wild-collected. The major exporters of bark include Cameroon, Madagascar, Equatorial Guinea, and Kenya. Groupe Fournier of France and Indena of Italy produce 86% of the world's bark extract, both for their own products and for the free market. Worldwide exports of dried bark in 2000 have been estimated at 1350-1525 metric tons per year, down from its peak of 3225 tons in 1997. Bark extracts (6370-7225 kg per year) are worth an estimated $4.36 million US dollars per year. In 2000, Plantecam, the largest bark exporter in Africa, closed its extraction factory in Cameroon, due to complex ecological, social, and economic factors. Wild collection is no longer sustainable (and probably never was) where harvest seriously affects morbidity and mortality rates of harvested populations. Since 1995, it has been included in CITES Appendix II as an endangered species. In this paper, alternatives to wild-collection to meet future market demand are investigated, including conservation practices, enrichment plantings, small- and large-scale production, and protection of genetic resources. The species is at the beginning of a transition from an exclusively wild-collected species to that of a cultivated medicinal tree. PMID- 14522427 TI - Hypolipidemic and hypocholesterolemic effect of argan oil (Argania spinosa L.) in Meriones shawi rats. AB - The potential health benefits of various dietary oils in relation to cardiovascular disease and cancer are recently receiving considerable attention. The main proposal of this study is to investigate the effect of dietary argan oil, obtained from seeds of Argania spinosa L. (Sapotaceae) endemic from Morocco, on serum lipids composition. Hyperlipidemia was induced by high calorie and cholesterol (HCC) diet administration in 16 rats (Meriones shawi, a rodent of the Gerbillideae family). Eight rats were treated with argan oil (1ml/100g weight) daily by oral route during 7 weeks (treated group). Control animals were also fed with HCC diet for 7 weeks. After 7-week treatment with argan oil, blood lipoproteins were significantly reduced. Total cholesterol decreased with 36.67% (P<0.01), low density lipoprotein (LDL)-cholesterol in 67.70% (P<0.001), triglycerides in 30.67% (P<0.05) and body weight in 12.7% (P<0.05). High density lipoprotein (HDL)-cholesterol concentration remained unaltered. These results indicate the beneficial effect of argan oil in the treatment of the hyperlipidemia and hypercholesterolemia. This effect will be related with the polyunsaturated fatty acids and other constituents of studied oil. PMID- 14522428 TI - Anti-inflammatory effects of crude methanolic extract and fractions of Alchornea cordifolia leaves. AB - The leaves of Alchornea cordifolia were collected, identified, dried, and reduced to coarse powder and extracted with aqueous methanol. Using various solvent treatments, the powdered dried leaf was fractionated into five fractions, A1, A2, B, C, D and E. The fractions were subjected to phytochemical analysis to identify the biologically active constituents. The anti-inflammatory effects of crude methanolic extract (ME) of Alchornea cordifolia leaves and the five fractions were evaluated using egg-albumen-induced rat hind paw oedema as a model of inflammation. The crude extract was also subjected to acute toxicity test. Fraction A2, which exhibited the most promising anti-inflammatory effect, was also subjected to analgesic and ulcerogenic tests. Phytochemical analysis of the extracts showed the presence of terpenes, sterols, flavonoids, tannins, carbohydrates, glycosides, saponins and traces of alkaloids. The LD(50) of the aqueous ME was found to be 1131.4 mg/kg. The crude ME (50 mg/kg) gave anti inflammatory activity which was significant (P<0.05) at all the observation times (1-3h). The different solvent fractions exhibited varying degrees of anti inflammatory activities, with terpenoid fraction (A2) and the tannin-containing multi-component fraction (D) showing very high and significant (P<0.01) activity at 100mg/kg, with percentage inhibition of oedema value of 87.69 each. In conclusion, the aqueous ME of Alchornea cordifolia leaves could be beneficial in the management of different inflammatory disease states. Its anti-inflammatory activity may not be attributed only to the terpenoid content. PMID- 14522429 TI - Evaluation of the anti-asthmatic property of Asystasia gangetica leaf extracts. AB - The leaf of Asystasia gangetica T. Adams (Acanthaceae) is used in many parts of Nigeria for the management of asthma. This study was aimed at investigating the anti-asthmatic property of hexane, ethylacetate, and methanol extracts of the leaves of Asystasia gangetica, obtained by successive sohxlet extraction. The results indicated that the extracts did not exhibit contractile or relaxant activity in isolated tissue preparations; however, they inhibited the contraction evoked by spasmogens; the IC(50) were calculated, where possible. The extracts relaxed histamine-precontracted tracheal strips in the following degree of potency-ethylacetate extract>hexane extract=methanol extract. The extracts also exhibited anti-inflammatory activity in the order of magnitude-methanol extract>hexane extract>ethylacetate extract. Acute toxicity test estimated an i.p. LD(50) of 2150 mg/kg in mice for methanol extract while phytochemical screening showed the presence of carbohydrates, proteins, alkaloids, tannins, steroidal aglycones, saponins, flavonoids, reducing sugars, and triterpenoids, with the methanol extract having the highest number of constituents. The study justified the use of the leaf of Asystasia gangetica in the management of asthma in Nigerian folk medicine. PMID- 14522430 TI - Radical-scavenging effects of Aloe arborescens Miller on prevention of pancreatic islet B-cell destruction in rats. AB - We evaluated the possible scavenging effects of Aloe arborescens Miller var. natalensis Berger (Kidachi aloe in Japanese) on free radicals generated by streptozotocin (Sz) or alloxan (Ax). The components of Kidachi aloe were added to a reaction system in which .OH radicals derived from Sz or Ax as pancreatic islet B-cell toxins and hypoxanthine-xanthine oxidase (HX-XO)-derived O(2) radicals destroy isolated islet B-cells, and we observed its preventive effects. The Kidachi aloe components inhibited the destruction of rat pancreatic islet B-cells by Sz, Ax or HX-XO. These components were prepared in the form of a freeze-dried powder of the boiled leaf skin of Kidachi aloe, and measurement of 1,1-diphenyl-2 picrylhydrazyl (DPPH) radical-scavenging activity showed higher radical scavenging activity in this boiled leaf skin powder than the non-boiled leaf skin powder.Furthermore, HPLC chromatograms of the "Boiled leaf skin powder" were similar to those of commercially available aloin (barbaloin content: approximately 20%). Therefore, the main component may be a phenol compound. In addition, the phenolic fraction of the Boiled leaf skin contained large amounts of 2'-O-p-coumaroylaloesin and 2'-O-feruloylaloesin, which have higher DPPH radical-scavenging activity than barbaloin. These results suggest that the action mechanism of Kidachi aloe Boiled leaf skin components, which prevent destruction of the pancreatic islets by specific pancreatic islet toxins such as Sz, Ax, and HX-XO, involves inhibition of free radical-scavenging effects, and may be associated with a thermostable low molecular component. The co-existence of Kidachi aloe-derived 2'-O-p-coumaroylaloesin, 2'-O-feruloylaloesin, and aloin may result in the potentiation of radical-scavenging activity. PMID- 14522431 TI - Induction of insulin secretion by a component of Urtica dioica leave extract in perifused Islets of Langerhans and its in vivo effects in normal and streptozotocin diabetic rats. AB - The blood glucose lowering effect of Urtica dioica (Stinging Nettle) as a medicinal plant has been noted in old writings such as those of Avicenna. Recently, there has also been other investigators that indicated the hypoglycemic effect of Urtica dioica. But so far, the mechanism of this effect has not been deduced. In this report, a perifusion system is arranged in which an exact number of Langerhans Islets were exposed to several fractions of extracts of Urtica dioica by TLC. The active ingredient fraction named F(1), caused a marked increase in insulin secretion. A simultaneous assay of glucose showed that the increase in insulin level was associated with a decrease in glucose level. Furthermore, the active component of Urtica dioica was found to increase the insulin content of blood sera in normal and streptozotocin diabetic rats that were injected intraperitoneally (i.p.) with the active ingredient of the extract. The in vivo studies presented in this report show that not only an increase in insulin level of blood sera was observed in rats after 30 min from the initial point of injection but a simultaneous decrease of blood sugar was detected when similar sera was tested for glucose. The increase in insulin level was six times during the 120 min of our determination. The decrease in blood sugar was found to be similar both in the level and time of initiation. On the basis of our findings, we assume that F(1) is the active ingredient of plant leaves extract. The results show that the blood lowering effect of the extract was due to the enhancement of insulin secretion by Langerhance Isletes. PMID- 14522432 TI - Onset of puberty and ovarian steroidogenesis following adminstration of methanolic extract of Cuscuta reflexa Roxb. stem and Corchorus olitorius Linn. seed in mice. AB - The effect of methanolic extract (ME) of Cuscuta reflexa stem Roxb. and Corchorus olitorius Linn. seed on the onset of reproductive maturity and the ovarian steroidogenesis was studied by means of biochemical techniques. ME of Cuscuta reflexa stem and Corchorus olitorius seed treatment causes a remarkable delay in sexual maturation as evidenced by the age at vaginal opening and appearance of first estrus (cornified smear). The same treatment also results in a significant diminution of Delta(5)-3beta-hydroxysteroid dehydrogenase (HSD) and glucose-6 phosphate dehydrogenase (G-6-PD) activity along with a reduction in the weight of ovary, uterus and pituitary. On the basis of above data, it is assumed that the probable cause of delayed maturation in ME of Cuscuta reflexa stem and Corchorus olitorius seed treated mice is due to the suppressed ovarian steroidogenesis. PMID- 14522433 TI - Effect of Hibiscus rosa sinensis Linn. ethanol flower extract on blood glucose and lipid profile in streptozotocin induced diabetes in rats. AB - Blood glucose and total lipid levels were determined in streptozotocin induced diabetic rats after oral administration of an ethanol flower extract of Hibiscus rosa sinensis. A comparable hypoglycemic effect was evidenced from the data obtained after 7 and 21 days of oral administration of the extract and glibenclamide. Maximal diminution in blood glucose (41-46%) and insulin level (14%) was noticed after 21 days. The extract lowered the total cholesterol and serum triglycerides by 22 and 30%, respectively. The increase in HDL-cholesterol was much higher (12%) under the influence of the extract as compared to that of glibenclamide (1%). The hypoglycemic activity of this extract is comparable to that of glibenclamide but is not mediated through insulin release. Other possible mechanisms are discussed. PMID- 14522434 TI - Anti-inflammatory and analgesic properties of the leaf extracts and essential oil of Lavandula angustifolia Mill. AB - Extracts obtained from the leaves of Lavandula angustifolia Mill. (Lamiaceae) are used in Iranian folk medicine as remedies for the treatment of various inflammatory diseases. For evaluation of its probable analgesic and anti inflammatory effects, hydroalcoholic extract, polyphenolic fraction and essential oil of the leaves of the herb were prepared and their analgesic effects were studied in mice using formalin and acetic acid-induced writhing tests. Carrageenan test in rats was used for assessment of anti-inflammatory activity of above-mentioned fractions. Results showed that while the hydroalcoholic extract (400-1600 mg/kg, p.o.) inhibited only the second phase of formalin test, the polyphenolic fraction (800 and 1600 mg/kg, p.o.) and essential oil (100 and 200 mg/kg, p.o.) suppressed both phases. In acetic acid-induced writhing test, polyphenolic fraction (400 and 800 mg/kg, p.o.) and essential oil (100 and 200 mg/kg, p.o.) reduced the number of abdominal constrictions. Essential oil at a dose of 200mg/kg also inhibited carrageenan-induced paw edema. Results of the present study confirm the traditional use of Lavandula angustifolia for the treatment of painful and inflammatory conditions and calls for further investigations to determine the active chemical constituent(s). PMID- 14522435 TI - Constituents of the Argentinian medicinal plant Baccharis grisebachii and their antimicrobial activity. AB - The resinous exudate of Baccharis grisebachii which is used to treat ulcers, burns, and skin sores in Argentina showed activity towards dermatophytes and bacteria. Two diterpenes, eight p-coumaric acid derivatives, and two flavones were isolated from the exudate and the structures elucidated by spectroscopic methods. 3-Prenyl-p-coumaric acid and 3,5-diprenyl-p-coumaric acid were active towards Epidermophyton floccosum and Trichophyton rubrum with MICs of 50 and 100 125 microg/ml, respectively. The diterpene labda-7,13E-dien-2beta,15-diol was active towards Epidermophyton floccosum and Trichophyton rubrum with MICs of 12.5 microg/ml while the MIC against Microsporum canis and Trichophyton mentagrophytes was 25 microg/ml. The diterpene was also active towards Microsporum gypseum with a MIC of 50 microg/ml, and showed inhibition in both Staphylococcus aureus (methicilline resistant and sensible strains) with MICs of 125 microg/ml. The results support the use of Baccharis grisebachii in Argentinian traditional medicine. PMID- 14522436 TI - The development of an in vitro screening strategy for topically applied products. AB - The objective of this study was the pharmaco-toxicological understanding of the constituents of an authenticated herbal mixture. The mixture was prepared by maceration in ethanol and subsequent dilution to produce a topically applied lotion, for which the intended target conditions are psoriasis and eczema. A three-tiered in vitro screening strategy was adopted for evaluating this product, comprising cytotoxicity assays; mutagenicity screening and therapeutic evaluation. Viability assays performed with dilutions of both the herbal concentrate and final product on organotypic cell lines indicated that neither preparation acted as an irritant. Genotoxicity screening using six strains of Salmonella typhimurium showed no mutagenic potential, and furthermore significant anti-microbial activity was evident. Therapeutic evaluation involved assessing the antioxidant potential of the extract, which can be correlated to an anti inflammatory effect. Nitroblue-tetrazolium (NBT) assay results indicate that the extract can reduce superoxide anion generation by 45%. The extract also increased cell viability on exposure to hydrogen peroxide by 28%, illustrating its dismutation potential. A 3-D skin culture system, EpiDerm, released 3000 microg/ml upon exposure to the extract, implying that the components enhance arachidonic acid metabolism. Overall, it may be concluded that the herbal extract is sufficiently non-toxic for dermal application and possesses anti-inflammatory activity. PMID- 14522437 TI - Pharmacological effect and toxicity of alkaloids from Gelsemium elegans Benth. AB - Gelsemium elegans Benth. (Loganiaceae), a toxic plant indigenous to southeastern Asia, is well known among hilltribes as an effective means for committing suicide. A crude alkaloidal fraction from its leaves was isolated by conventional acid-base extraction. Tests in animals showed that the alkaloids exerted analgesic and anti-inflammatory effects. At a lethal dose, the alkaloids produced violent clonic convulsions that led to respiratory failure. Since the convulsions could be prevented by pentobarbital or diazepam and potentiated by reserpine, it is postulated that the alkaloids act centrally against GABA action. PMID- 14522438 TI - Toxicity of a subchronic treatment with hydroalcoholic crude extract from Solanum grandiflorum (Ruiz et Pav) in rats. AB - The toxic effect of the hydroalcoholic crude extract (HACE) from the fruits of the Solanum grandiflorum was evaluated following subchronic treatment (p.o.) in Wistar rats for 65 days using the dose of 2 mg/kg. The LD(50) in mice was 50 mg/kg (p.o.). At the end of treatment, the consumption of water remained within the normal patterns. On the other hand, the consumption of food had 40% reduction in the period between the 18th and the 40th day after the beginning of the study. The diuresis showed a wide variation in volume which started at the 8th day of treatment until the end of the study. This finding was correlated with macroscopic observations of the kidneys which presented nephromegaly (70%) when compared to the control group. Moreover, for the histopathological study, it was observed degeneration and glomerular necrosis (100%), tubular lesions (80%), and hypercellular lesions (100%) in the treated group. The ponderal development of the treated group was increased compared to the control group. The weight of some of the organs, including kidneys, lungs, and pancreas, was also increased in the treated group (P<0.05, Student "t" test). These data suggest a nephrotoxicity following a subchronic treatment of the hydroalcoholic crude extract from Solanum grandiflorum. PMID- 14522439 TI - Coagulant and anticoagulant activities in Jatropha curcas latex. AB - Jatropha curcas Linn. (Euphorbiaceae), a medicinal plant commonly grown in the Tropics, is traditionally used as a haemostatic. Investigation of the coagulant activity of the latex of Jatropha curcas showed that whole latex significantly (P<0.01) reduced the clotting time of human blood. Diluted latex, however, prolonged the clotting time: at high dilutions, the blood did not clot at all. This indicates that Jatropha curcas latex possesses both procoagulant and anticoagulant activities. Prothrombin time (PT) and activated partial thromboplastin time (APTT) tests on plasma confirm these observations. Solvent partitioning of the latex with ethyl acetate and butanol led to a partial separation of the two opposing activities: at low concentrations, the ethyl acetate fraction exhibited a procoagulant activity, while the butanol fraction had the highest anticoagulant activity. The residual aqueous fraction had no significant effect on the clotting time of blood and the PT but slightly prolonged the APTT. PMID- 14522440 TI - Antitumour activity of Bauhinia variegata on Dalton's ascitic lymphoma. AB - The antitumour activity of the ethanol extract of Bauhinia variegata (EBV) has been evaluated against Dalton's ascitic lymphoma (DAL) in Swiss albino mice. A significant enhancement of mean survival time of EBV-treated tumour bearing mice was found with respect to control group. EBV treatment was found to enhance peritoneal cell counts. After 14 days of inoculation, EBV is able to reverse the changes in the haemotological parameters, protein and PCV consequent to tumour inoculation. PMID- 14522441 TI - N'Dribala (Cochlospermum planchonii) versus chloroquine for treatment of uncomplicated Plasmodium falciparum malaria. AB - The aim of this work was to assess the efficacy of oral N'Dribala (tuberous roots decoction of Cochlospermum planchonii Hook) treatment versus chloroquine in non severe malaria. The study included 85 patients with uncomplicated Plasmodium falciparum infection in Banfora, Burkina Faso. Forty-six patients that received N'Dribala beverage were compared to 21 patients treated with chloroquine. All patients were monitored with clinical examination and a parasitemia control by Giemsa-stained thick films. N'Dribala appeared safe and statistically as efficient as chloroquine for the treatment of uncomplicated Plasmodium falciparum malaria. At day 5 (D5), 57% of chloroquine-treated and 52% of N'Dribala-treated patients were cured with no detectable parasitemia (parasite density (Pd): 0) and more than 90% of whole patients were asymptomatic. N'Dribala is easily available in this country, cheap, without significant side effects and efficient with a clearly demonstrated activity on Plasmodium falciparum blood stages. This study enhances the traditional use of the Cochlospermum planchonii as alternative therapy for treatment of non-severe malaria. PMID- 14522442 TI - Acute and subacute toxicities of the saponin mixture isolated from Schefflera leucantha Viguier. AB - Acute toxicity of the bronchodilator saponin mixture isolated from Schefflera leucantha Viguier leaves was investigated in comparison with the methanol and the water extract of this plant. Oral doses of 5000 mg/kg of the methanol extract, the water extract and the saponin mixture did not produce mortality or significant changes in the general behavior and gross appearance of internal organs of rats. Subacute toxicity of the saponin mixture was evaluated with the dose of 1000 mg/kg, orally for 14 days. An extra group (satellite group) was given saponin mixture and kept for a further 14 days after treatment. All animals did not show signs of toxicity during the experimental period. Liver weights of the saponin-treated and the satellite male groups were higher whereas testis weight were lower than those of the control group which received distilled water. However, the histological examination of various organs revealed that there were no differences between the control and the treated rats. BUN, Cr, AST, ALT and ALP levels increased in saponin-receiving rats. It is possible that the saponin mixture directly impacts on the liver and the kidney functions. PMID- 14522443 TI - Anti-inflammatory and antinociceptive activity assessment of plants used as remedy in Turkish folk medicine. AB - Ethanolic and aqueous extracts from seven plant species used in Turkish traditional medicine were evaluated for in vivo anti-inflammatory and antinociceptive activities; Helleborus orientalis Lam. roots and herbs, Juglans regia L. leaves, Laurocerasus officinalis Roemer leaves, Nerium oleander L. dried and fresh flowers and leaves, Rhododendron ponticum L. leaves, Rubus hirtus Walds. et Kit aerial parts and Rubus sanctus Schreber aerial parts and roots. All the plant extracts, except the aqueous extract of Rubus hirtus, were shown to possess significant antinociceptive activity in varying degrees against p benzoquinone-induced abdominal contractions in mice. However, only the ethanolic extracts of Helleborus orientalis roots, Juglans regia leaves, Laurocerasus officinalis leaves, Nerium oleander dried and fresh flowers, and Rhododendron ponticum leaves exhibited potent anti-inflammatory activity against carrageenan induced hind paw edema model in mice without inducing any gastric damage. Results of the present study confirmed the folkloric claim that all the selected materials to possess potent antinociceptive and anti-inflammatory activity. PMID- 14522445 TI - Anti-inflammatory effects of an herbal medicine (Xuan-Ju agent) on carrageenan- and adjuvant-induced paw edema in rats. AB - Xuan-Ju agent is an herbal formula containing aqueous extract of Formica fusca, Herba epimedii, Fructus cnidii, and Fructus lycii, all of which are reputed for their beneficial effects in the treatment of the immunodeficient diseases such as rheumatoid arthritis. We performed a study on the anti-inflammatory effects of this agent using carrageenan- and adjuvant-induced paw edema in rats. Xuan-Ju agent showed a marked inhibitory effect on edema in two models of inflammation in rats, at the dose of 0.20, 0.40 and 0.80g/kg. Based on this study, Xuan-Ju agent is considered to be a potentially useful drug suitable for further evaluation for rheumatoid arthritis. PMID- 14522444 TI - Pharmacological approach to the pro- and anti-inflammatory effects of Ranunculus sceleratus L. AB - Ranunculus sceleratus is a widespread species with unique toxicological and pharmacological activities. The present study seeks to assess this species' ability, both in vitro and in vivo, to modulate processes involved in inflammations. To this end, different extracts from the aerial parts of the plant were tested in several models of acute inflammation induced by tetradecanoylphorbol acetate (TPA), arachidonic acid (AA), and carrageenan, as well as in two models of delayed hypersensitivity induced by oxazolone and dinitrofluorobencene (DNFB). The extracts were also assayed in models of eicosanoid and elastase release by intact cells. When tested in vivo, all of the extracts showed anti-inflammatory or neutral effects. In vitro, non-polar extracts of this species were able to inhibit eicosanoid production, whereas polar extracts enhanced the synthesis of 5(S)-HETE, LTB(4) and 12(S)-HHTrE. The hypothesis of a "counter-irritant" mechanism of action has thus been proposed and is also discussed herein. PMID- 14522446 TI - Effects of a caffeine-free Cola nitida nuts extract on elastase/alpha-1 proteinase inhibitor balance. AB - In an infection, polymorphonuclear neutrophils (PMN) become activated and they produce oxidizing compounds and elastase in the extracellular medium. Alpha-1 proteinase inhibitor (alpha1PI), a protease inhibitor which is inactivated by oxidants, is the main endogenous inhibitor of elastase helping to limit excessive elastase activity. This study evaluates the ability of a plant extract, Cola nitida nuts, to protect alpha1PI from inactivation by oxidizing compounds as reactive oxygen species. On the one hand, we have evaluated the direct effect of cola nut extract on neutrophil elastase, and on the H(2)O(2) and myeloperoxidase (MPO)-H(2)O(2) system via cell-free systems. Results showed that cola nut extract scavenges H(2)O(2) and therefore protects alpha1PI from HOCl which is produced from the MPO-H(2)O(2) system. Experiments also showed that cola extract has the capacity to limit elastase activity. On the other hand, we have worked on cellular systems including isolated PMN with the aim to study the effect of cola extract on PMN metabolism. PMN were stimulated with PMA, calcium ionophore or fMLP. Each stimulant possesses its own stimulation pathway. According to the inhibitory concentration obtained at 50%, the results on cellular systems led to the conclusion that cola extract can reduce elastase liberation from PMN. It can then be concluded that cola nut extract can protect alpha1PI from inactivation, and has an effect both on elastase liberation and elastase activity. The cola nut extract effect is rather biased towards a reduction in elastase release, thus limiting the injurious effects caused by this enzyme. PMID- 14522447 TI - Angiotensin converting enzyme inhibitory phenylpropanoid glycosides from Clerodendron trichotomum. AB - The stems of Clerodendron trichotomum have been traditionally used for treatment of hypertension in far East Asia including China, Korea, and Japan. Bioassay guided fractionation and purification of the EtOAc-soluble extract of Clerodendron trichotomum afforded acteoside (1), leucosceptoside A (2), martynoside (3), acteoside isomer (4), and isomartynoside (5). The angiotensin converting enzyme (ACE) activities were significantly inhibited by the addition of these phenylpropanoid glycosides (1-5) in a dose-dependent manner of which IC(50) values were 373+/-9.3 microg/ml, 423+/-18.8 microg/ml, 524+/-28.1 microg/ml, 376+/-15.6 microg/ml, 505+/-26.7 microg/ml, respectively. These results suggest that the antihypertensive effect of Clerodendron trichotomum may be, at least in part, due to ACE inhibitory effect of phenylpropanoid glycosides. PMID- 14522448 TI - Immunomodulatory activities of Clausena excavata Burm. f. wood extracts. AB - In vitro immunomodulatory activities of aqueous extract, acetone extract and the Thai folklore extract of Clausena excavata Burm. f. on mouse immune system were investigated. The phagocytic activity of macrophages and splenocyte proliferation in the absence and presence of mitogens (lipopolysaccharide, LPS) or pokeweed mitogen, PWM) were assayed. The aqueous extract exhibited the maximum effect on both respiratory burst response and lysosomal enzyme activity more than the acetone extract and the Thai folklore extract indicating effective phagocytic activation. For splenocyte proliferation assay, the Thai folklore extract with LPS gave the maximum activity higher than that with PWM, suggesting specificity towards B cell proliferation through T cell independent pathway the same as LPS. The present study revealed the immunomodulating activity, which could be explained the traditional use of this plant in Thailand. PMID- 14522449 TI - Toxicological investigation of aqueous-methanolic extract of the calyces of Hibiscus sabdariffa L. AB - The aqueous fraction of an aqueous-alcoholic extract of Hibiscus sabdariffa L. calyces was given to Wistar albino rats (150-200g) orally as drugs to study the toxicity of the extract. The rats, which were fed with commercial diet and water ad libitum, were divided into six groups of four rats each. Treatments 1 through 6 received 0, 1, 3, 5, 10 and 15 doses of 250 mg/kg each, respectively; the control group received physiological saline (0.9% NaCl). Results of the studies showed that the levels of serum aspartate aminotransferase (AST) and alanine amino transferase (ALT) were significantly (P<0.05) increased in all the treatments compared with the control group. However, the serum levels of alkaline phosphatase, and lactate dehydrogenase were not significantly (P>0.05) affected. Only the group with 15 doses had their serum level of albumin significantly (P<0.05) increased. However, the results of histopathological studies showed that both the livers and hearts gave no pathological features for all the treatments. The results showed that prolong usage of this extract at 15-dose level could cause liver injury while the effect was mild at small dose levels (1-10). Though the average consumption of 150-180 mg/kg per day appears safe, the extract should be taken with caution bearing in mind that higher doses could affect the liver. PMID- 14522450 TI - Chemical composition and antifungal activity of essential oils of seven Moroccan Labiatae against Botrytis cinerea Pers: Fr. AB - Essential oils of seven Moroccan Labiatae were chemically analysed by GC-MS and evaluated for their in vitro antifungal activity against Botrytis cinerea. Among them, Origanum compactum and Thymus glandulosus greatly inhibited the growth of the mycelium. The inhibition of Botrytis cinerea was 100% for both oils at 100 ppm, while the IC(50)s were 35.1 and 79.2 ppm, respectively. Mentha pulegium exhibited moderate activity at 250 ppm since the inhibition of the mycelial growth was 58.5% and the IC(50) was 233.5 ppm. The main constituents of the studied oils were also examined. Thymol and carvacrol that are the two main constituents of Thymus glandulosus and Origanum compactum exhibited the strongest antifungal activity with 100% of inhibition at 100 ppm, respectively. PMID- 14522451 TI - Investigations of anti-inflammatory and antinociceptive activities of Piper cubeba, Physalis angulata and Rosa hybrida. AB - The anti-inflammatory activities of Piper cubeba (fruit), Physalis angulata (flower) and Rosa hybrida (flower) were determined by carrageenan-induced paw edema, arachidonic acid-induced ear edema and formaldehyde-induced arthritis in mice. The anti-allergic and analgesic activities of these plants were also studied by using 2,4-dinitrofluorobenzene (DNFB)-induced contact hypersensitivity reaction (type IV) and hot plate test in mice, respectively. These plant extracts clearly exhibited inhibitory effects against acute and subacute inflammation by oral administration (200 mg/kg). Also, administration (200 mg/kg, p.o.) of plant extracts for 1 week significantly inhibited type IV allergic reaction in mice (P<0.05). Rosa hybrida showed an analgesic effect against hot plate-induced thermal stimulation at a dose of 200 mg/kg. These results provide support for the use of Rosa hybrida in relieving inflammatory pain, and insight into the development of new agents for treating inflammatory diseases. PMID- 14522453 TI - Anti-adhesion therapy of bacterial diseases: prospects and problems. AB - The alarming increase in drug-resistant bacteria makes a search for novel means of fighting bacterial infections imperative. An attractive approach is the use of agents that interfere with the ability of the bacteria to adhere to tissues of the host, since such adhesion is one of the initial stages of the infectious process. The validity of this approach has been unequivocally demonstrated in experiments performed in a wide variety of animals, from mice to monkeys, and recently also in humans. Here we review various approaches to anti-adhesion therapy, including the use of receptor and adhesin analogs, dietary constituents, sublethal concentrations of antibiotics and adhesin-based vaccines. Because anti adhesive agents are not bactericidal, the propagation and spread of resistant strains is much less likely to occur than as a result of exposure to bactericidal agents, such as antibiotics. Anti-adhesive drugs, once developed, may, therefore, serve as a new means to fight infectious diseases. PMID- 14522454 TI - Acute inflammatory response induced by Helicobacter pylori in the rat air pouch. AB - Infection by Helicobacter pylori elicits persistent neutrophil infiltration in the gastric mucosa and stimulates the release of substances that may contribute to the establishment of gastritis. In this study, we used the rat air pouch model to evaluate the acute inflammatory response to H. pylori, in vivo. A pronounced neutrophil infiltration was observed 6 h and 12 h after the injection of H. pylori into the air pouch. Strains with different genotypes were able to induce cellular influx. This response was dependent upon the amount of bacteria injected and still occurred when heat-killed bacteria were employed. An increase in prostaglandin E(2) levels was observed, indicating that H. pylori induced cyclooxygenase 2 in this model. The production of interleukin-1 beta and tumor necrosis factor-alpha by leukocytes was also enhanced, suggesting that this model may be useful for studying the direct activation of neutrophils by H. pylori in vivo. PMID- 14522455 TI - Novel applications of feather tip extracts from MDV-infected chickens; diagnosis of commercial broilers, whole genome separation by PFGE and synchronic mucosal infection. AB - Marek's disease virus (MDV) productive replication occurs in the feather follicle epithelium and the feather tips are valuable both for research and disease diagnosis. Three novel applications of feather tip extracts are described now: (A). As a source of DNA for amplifying either MDV and/or ALV-J. In two clinical situations a marked advantage was obtained compared to blood and organs; in broiler breeder flocks with a mixed MDV and ALV-J infection, and in young broilers with neurological Marek's disease (MD). (B). Separation of the large ( approximately 200 kbp) MDV genome directly from the infected chickens. Using pulsed field gel electrophoresis, the DNA extracted from tumors or feather tips was separated and hybridized to a 132 bp tandem repeat MDV probe. Compared to 2/55 polymerase chain reaction (PCR) positive tumor samples, 15/61 feather tip extracts contained whole MDV genomes. (C). Experimental MDV infection was induced by the mucosal route by dripping feather tip extract to the eye and mouth of the bird. That attempted to reproduce the native infection process, however the use of extracts, instead of dry feather dust was a compromise, aimed to synchronize the infection. In one trial, tumors were induced 6 weeks after dripping day-old broilers, while in another, feather tips were PCR positive 16 days after dripping of 2-month-old layers. PMID- 14522456 TI - Role of M3 protein in the adherence and internalization of an invasive Streptococcus pyogenes strain by epithelial cells. AB - Streptococcus pyogenes utilizes multiple mechanisms for adherence to and internalization by epithelial cells. One of the molecules suggested of being involved in adherence and internalization is the M protein. Although strains of the M3 serotype form the second largest group isolated from patients with severe invasive diseases and fatal infections, not much information is known regarding the interactions of M3 protein with mammalian cells. In this study we have constructed an emm3 mutant of an invasive M3 serotype (SP268), and demonstrated that the M3 protein is involved in both adherence to and internalization by HEp-2 cells. Fibronectin promoted both adherence and internalization of SP268 in an M3 independent pathway. Utilizing speB and speB/emm3 double mutants, it was found that M3 protein is not essential for the maturation of SpeB, as was reported for the M1 protein. Increased internalization efficiency observed in both the speB and emm3/speB mutants suggested that inhibition of S. pyogenes internalization by SpeB is not related to the presence of an intact M3 protein. Thus, other proteins in SP268, which serve as targets for SpeB activity, have a prominent role in the internalization process. PMID- 14522457 TI - Developing subunit immunogens using B and T cell epitopes and their constructs derived from the F1 antigen of Yersinia pestis using novel delivery vehicles. AB - Yersinia pestis is the etiological agent of pneumonic and bubonic plague. As the currently licensed vaccines for plague have their own limitations, there is a need for a rational and more effective form of a subunit vaccine to combat both forms of the disease. Newer methods of antigen delivery coupled with adjuvant offer an alternative approach toward a plague vaccine. In order to develop a new generation vaccine against plague, we chose an immunodominant, outer membrane capsular protein, F1 of Y. pestis. The immunogenicity of the peptide sequences, predicted to possess B (three sequences, B1, B2 and B3) and T (two sequences, T1 and T2) cell determinants, was studied in a murine model with different genetic backgrounds, using alhydrogel and liposomes as delivery vehicles. All the peptide sequences are immunogenic in all mouse strains and showed primary and secondary immune response. B2 peptide was found to be most immunogenic, followed by B1 and B3 peptides. Chimeras made between B and T structures proved highly immunogenic and the antibody levels are comparable with native F1 antigen, thereby proving that T1 and T2 are helper sequences. Interestingly, the liposome mode of immunization was found to be more immunogenic and generated higher affinity antibodies than the alum-based preparation. Immunization using a mixture of all the peptides further proved B2 to be immunodominant. The IgG isotype profile showed predominance of IgG1, IgG2b followed by IgG2a for all the formulations irrespective of mode of antigen delivery. Lymphocyte proliferation of spleen cells primed in vivo with peptides, B-T conjugates and F1 antigen followed by in vitro stimulation with these antigens in soluble (medium) and particulate (liposome) form, showed dose-dependent stimulation of T cells, while B-T constructs showed a higher stimulation index, comparable to F1 antigen. The liposome mode of antigen presentation showed higher lymphoproliferation of spleen cells. Of all the peptides tested, T1 and T2 sequences showed the highest stimulation indices. The pattern of cytokine levels was in the following order: interferon-gamma>interleukin-2>interleukin-4. In vivo protective studies of the B T conjugates revealed that B1T1 and a mixture of conjugates showed a survival rate of 10 days. Thus, the study highlights the importance of B and T cell epitopes as peptide-based immunogens, being a serious alternative for plague vaccine. PMID- 14522458 TI - Specific activation of CMV-primed human T lymphocytes by cytomegalovirus pp65 expressed in fission yeast. AB - Threatening virus infections constantly illustrate the growing need for novel vaccines that specifically induce efficient T cell-mediated immune responses. In this study, we used a human whole blood assay to determine the activation of antigen-specific human T lymphocytes by a viral antigen of human cytomegalovirus (HCMV). The major HCMV tegument protein pp65, recombinantly expressed in fission yeast (Schizosaccharomyces pombe), specifically activated antigen-specific CD4- and CD8-positive memory T cells in blood of HCMV seropositive donors. Moreover, the immune response against recombinant pp65, in particular that of CD8 class I major histocompatibility complex-restricted cytotoxic T cells, was similar to the response against the intact HCMV. Since fission yeast cells per se did not activate a significant number of human T lymphocytes ex vivo, the system described here might represent a novel approach in vaccine development as well as in the identification of vaccine candidates directly from human whole blood. PMID- 14522459 TI - Evaluation and comparison of random amplification of polymorphic DNA, pulsed field gel electrophoresis and ADSRRS-fingerprinting for typing Serratia marcescens outbreaks. AB - Amplification of DNA fragments surrounding rare restriction sites (ADSRRS fingerprinting) is a novel assay based on suppression of polymerase chain reaction (PCR). This phenomenon allows the amplification of only a limited subset of DNA fragments, since only those with two different oligonucleotides ligated at the ends of complementary DNA strands are amplified in the PCR. The DNA fragments can be easily analyzed on polyacrylamide gels, stained with ethidium bromide. We have implemented this method using a set of clinical Serratia marcescens isolates from three outbreaks ongoing in the Public Hospital in Gdansk (Poland). Clustering of ADSRRS-fingerprinting data matched epidemiological, microbiological, random amplification of polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) data. Based on this study, we found that there is at least a similar power of discrimination between the present 'gold-standard' PFGE and the novel method, ADSRRS-fingerprinting. Although the ADSRRS-fingerprinting method may appear to be more complex than the RAPD technique, we found it fast and reproducible. PMID- 14522460 TI - Restoration of mycobacterial antigen-induced proliferation and interferon-gamma responses in peripheral blood mononuclear cells of tuberculosis patients upon effective chemotherapy. AB - Peripheral blood mononuclear cells (PBMC) were obtained from culture-proven tuberculosis (TB) patients before and after 2 and 6 months of chemotherapy with a multi-drug regimen. PBMC were tested for cellular responses in antigen-induced proliferation and interferon-gamma (IFN-gamma) assays in response to complex mycobacterial antigens (whole cell Mycobacterium bovis BCG and M. tuberculosis, cell walls and short-term culture filtrate [ST-CF] of M. tuberculosis), fractionated ST-CF antigens (fractions F1-F10) and ESAT-6. The responses in TB patients before anti-TB treatment were low (median stimulation index (SI)=1-7, median delta IFN-gamma=0-12 U ml(-1), and percent responders=13-67%) to all the antigenic preparations. Following the administration of anti-TB chemotherapy for 2 months, there were significant (P<0.05) improvements in the cellular responses (median SI=9-76, median delta IFN-gamma=3-70 U ml(-1), and percent responders=33 100%) to most of the antigenic preparations tested. However, concanavalin A induced proliferation responses of PBMC from the same patients before and after 2 months of chemotherapy were high and comparable (median SI=101 and 114, respectively, P>0.05, 100% responders). A further increase in IFN-gamma responses (median delta IFN-gamma=14-250 U ml(-1) and percent responders=43-100%) to mycobacterial antigens was observed in patients receiving chemotherapy for 6 months. Among the ST-CF fractions, F1 and F2 containing low molecular mass proteins resulted in the highest responses, whereas ESAT-6 showed responses comparable to these fractions only in a minority of the patients. HLA-DR typing of these patients showed heterogeneity in the expression of molecules encoded by HLA-DRB genes. These results show that effective chemotherapy restores cellular responses of TB patients to a large number of M. tuberculosis antigens, which could be useful in monitoring the efficacy of anti-TB treatment. PMID- 14522461 TI - Heteroduplex mobility assay for the identification of Listeria sp and Listeria monocytogenes strains: application to characterisation of strains from sludge and food samples. AB - One hundred and ten Listeria sp. isolates from sewage sludge were identified according to phenotypic and genotypic methods. The Listeria sp. strains isolated from five types of sludge from three sewage treatment plants in Angers (France) and the surrounding area included L. monocytogenes (55.5%), L. innocua (29.1%), L. seeligeri (13.6%) and L. welshimeri (1.8%). The majority of L. monocytogenes strains belonged to serotypes 4b, 1/2b and 1/2a. Moreover, a heteroduplex mobility assay based on the 16S rRNA sequences was tested for its ability to identify the six species of the genus Listeria. This study, performed on 283 Listeria sp. strains from human, food and sewage sludge samples, showed that all the species were distinguishable from one another. L. innocua and L. seeligeri showed respectively three and two distinct banding patterns. Within L. monocytogenes, four groups (I-IV) were defined. The majority of food and environmental isolates were clustered in group I and it is noteworthy that group IV clustered epidemiologic isolates and strains belonging to serotypes 4b, 1/2a and 1/2b. PMID- 14522462 TI - Application of real-time PCR for quantitative detection of Campylobacter jejuni in poultry, milk and environmental water. AB - Campylobacter jejuni is a leading human food-borne pathogen. The rapid and sensitive detection of C. jejuni is necessary for the maintenance of a safe food/water supply. In this article, we present a real-time polymerase chain reaction (PCR) assay for quantitative detection of C. jejuni in naturally contaminated poultry, milk and environmental samples without an enrichment step. The whole assay can be completed in 60 min with a detection limit of approximately 1 CFU. The standard curve correlation coefficient for the threshold cycle versus the copy number of initial C. jejuni cells was 0.988. To test the PCR system, a set of 300 frozen chicken meat samples, 300 milk samples and 300 water samples were screened for the presence of C. jejuni. 30.6% (92/300) of chicken meat samples, 27.3% (82/300) of milk samples, and 13.6% (41/300) of water samples tested positive for C. jejuni. This result indicated that the real-time PCR assay provides a specific, sensitive and rapid method for quantitative detection of C. jejuni. Moreover, it is concluded that retail chicken meat, raw milk and environmental water are commonly contaminated with C. jejuni and could serve as a potential risk for consumers in eastern China, especially if proper hygienic and cooking conditions are not maintained. PMID- 14522463 TI - Pro-inflammatory effects of Burkholderia cepacia on cystic fibrosis respiratory epithelium. AB - Burkholderia cepacia causes pulmonary infection with high mortality in cystic fibrosis (CF) patients which is likely to involve interaction with respiratory epithelium. In this study the pro-inflammatory properties of B. cepacia were examined using a range of respiratory epithelial cell lines. B. cepacia and cell free culture supernatants were used to stimulate cell lines with (SigmaCFTE29o- and IB3) and without (A549) the CF transmembrane conductance regulator mutation (CFTR), together with corrected cell lines (C38 and S9). Interleukin (IL)-6 and IL-8, but not GM-CSF or IL-1beta, were released from all the cell lines whereas PGE(2) (prostaglandin E(2)) was released from the A549, IB3 and S9 cell lines only. Nuclear factor (NF)-kappaB activation preceded cytokine release and suppression of NF-kappaB activity diminished cytokine release. These studies indicated that B. cepacia secretory products are potent pro-inflammatory agents for respiratory epithelium and suggest functional CFTR is not required for cytokine or prostanoid responses. PMID- 14522464 TI - Hopping to the beat. Hop regulation of cardiac gene expression. AB - Hop is a recently discovered homeodomain only protein with atypical characteristics. Unlike other homeodomain proteins, Hop lacks DNA binding activity, although it localizes predominantly to the nucleus. During embryogenesis, Hop is expressed in the myocardium of the developing heart and it functions downstream of Nkx2-5, an early cardiac-specific transcription factor. One role of Hop is to modify the expression of cardiac-specific genes and thereby finely regulate heart development. This article reviews the discovery, characteristics, roles, and future perspectives of Hop. PMID- 14522465 TI - Role of blood mononuclear cells in recanalization and vascularization of thrombi: past, present, and future. AB - Recent experimental data suggest a role for blood-borne mononuclear cells, including bone marrow-derived endothelial progenitor cells (EPCs) in organization, vascularization, and recanalization of thrombi. Older studies have described in detail the in situ structural modifications accompanying these processes. The common themes are (a) involvement of pluripotent mononuclear cells, and (b) proteolytic and/or phagocytic activity of monocytes/macrophages (MCs/Mphs) in modifying the thrombus to a proangiogenic state. In addition to a nurturing function, MCs/Mphs may assist the engraftment of thrombus-trapped or incoming EPCs. The differences between the recanalization potency of venous and arterial thrombi suggest that in addition to the organization of the fibrin meshwork, the cellular composition of the venous thrombi may be responsible for the better recanalization seen in venous clots. These observations set the stage for therapeutic manipulation of organization and recanalization of thrombi, by increasing the MC/Mph and/or EPC content. PMID- 14522466 TI - DNA damage as a new emerging risk factor in atherosclerosis. AB - Atherosclerosis is the leading cause of illness and death in Western societies. It is now clear that the disease is a result of a chronic fibroproliferative inflammatory response of the arterial intima leading to the formation of atherosclerotic plaque. There is, however, strong experimental evidence for a "mutation theory of atherosclerosis," underlining the similarity between atherosclerotic and carcinogenic processes. This review provides an overview of the studies that support the role of genetic alterations in the disease. The demonstration of microsatellite instability and loss of heterozygosity in smooth muscle cells of human plaques suggests that genomic destabilization may play a pivotal role in atherosclerotic mechanisms. Furthermore, the use of accepted biomarkers of carcinogenic exposure-such as DNA adducts and cytogenetic end points-recently has provided evidence consistent with the view that somatic cell alterations are critical in atherogenic process. It follows that the study of DNA damage may provide new insights into the pathogenesis of atherosclerosis and lead to the development of novel therapeutic approaches. PMID- 14522467 TI - EG-VEGF and Bv8. a novel family of tissue-selective mediators of angiogenesis, endothelial phenotype, and function. AB - Angiogenic molecules are the focus of therapeutic efforts to promote new vessel development in ischemic or damaged tissue and, conversely, to inhibit endothelial cell growth and survival in proliferative disease. Two novel angiogenic mitogens have been characterized recently. Endocrine gland-derived vascular endothelial growth factor (EG-VEGF) and the mammalian homologue of Bombina variegata peptide 8 (Bv8) are endothelial cell mitogens and chemotactic factors with restricted expression profiles and selective endothelial cell activity. These highly related peptides share two cognate G-protein-coupled receptors that are homologous to the neuropeptide Y receptor. The identification of tissue-selective angiogenic factors raises the possibility that other secreted molecules in this class exist. The potential advantage of tissue-specific angiogenic therapeutics may be the reduction of systemic side effects. Additionally, these peptides or their receptors may be attractive targets for inhibition in several disorders. PMID- 14522468 TI - Novel approaches for the study of vascular assembly and morphogenesis in avian embryos. AB - Dynamic imaging of primary capillary bed formation in a warm-blooded embryo now is readily accomplished with the use of modern digital cameras, software, and instrumentation. The precise dynamic behavior of endothelial cells and their emergent vascular patterns are easily recorded and quantified in exquisite detail. As an example, we present data regarding vasculogenesis and vascular remodeling under normal and vascular endothelial growth factor-stimulated conditions, including corresponding computational analyses of endothelial cell behavior. The results show that the quail embryo provides an excellent experimental system to test reagents hypothesized to play a role in vascularization of tumors, engineered tissues, or wound sites. PMID- 14522469 TI - Molecular and functional maturation of the murine cardiac conduction system. AB - The rhythmic heartbeat reflects a highly coordinated electrical wave, initiated and propagated throughout the myocardium by the specialized cardiac conduction system (CCS). Conduction system abnormalities affect millions of individuals, with symptoms from merely bothersome palpitations to fainting or even sudden death. Despite years of investigation, knowledge of the genes and molecules that direct CCS development and function are incomplete. In 1977, Viragh and Challice published the first in a series of articles describing the morphologic development of the murine CCS. The framework established by those reports now can be revisited in light of recent data. Imaging the developing CCS-using genetically engineered mice combined with functional assays, including optical mapping of cardiac electrical excitation-reveal some of the earliest events in CCS formation and function. These findings build on the experiments of Viragh and Challice, leading to a more integrated picture of conduction system development in the mammalian embryo. PMID- 14522470 TI - Characterization and culture of human embryonic stem cells. AB - Human embryonic stem (ES) cells are cultured cell lines derived from the inner cell mass of the blastocyst that can be grown indefinitely in their undifferentiated state, yet also are capable of differentiating into all cells of the adult body as well as extraembryonic tissue. Detailed investigation of the properties of embryonal carcinoma cells of both the mouse and human as well as mouse and primate ES cells led to the initial isolation and subsequent culture of human ES cells. The methodologies that were developed to culture and characterize these cell lines have provided a template for the development of human ES cells. The existing data illustrate a number of important differences and similarities between human ES cells and the other cell lines. This review aims to provide a brief historic account of the development of the mammalian pluripotent stem cell field; describe how this led to the isolation, culture, and characterization of human ES cells; and discuss the potential implications of recent advances. PMID- 14522471 TI - Controlling the angiogenic switch: a balance between two distinct TGF-b receptor signaling pathways. AB - Biochemical studies in endothelial cells (ECs) and genetic studies in mice and humans have yielded major insights into the role of transforming growth factor beta (TGF-beta) and its downstream Smad effectors in embryonic vascular morphogenesis and in the establishment and maintenance of vessel wall integrity. These studies showed that TGF-beta signaling is of critical importance for normal vascular development and physiology. They also indicated the involvement of two distinct TGF-beta signaling cascades within ECs, namely the activin receptor-like kinase 5 (ALK5)-Smad2/3 pathway and the ALK1-Smad1/5 pathway. Aberrant TGF-beta signaling forms the basis for several vascular disorders such as hereditary hemorrhagic telengiectasia and primary pulmonary hypertension as well as neovascularization during tumorigenesis. This review describes the role of TGF beta in angiogenesis and some of the controversial issues concerning TGF-beta signaling through ALK1 and ALK5 in ECs. PMID- 14522472 TI - The clinical implications of endothelial dysfunction. AB - Defining new approaches for the prevention and treatment of atherosclerosis is an important priority. Recently, measurement of endothelial function in patients has emerged as a useful tool for atherosclerosis research. Risk factors are associated with impaired endothelial function, and clinical syndromes relate, in part, to a loss of endothelial control of vascular homeostasis. Recent studies have shown that the severity of endothelial dysfunction relates to cardiovascular risk. A growing number of interventions known to reduce cardiovascular risk have been shown to improve endothelial function. This work suggests that studies of endothelial function could be used in the care of patients and as a surrogate marker for the evaluation of new therapeutic strategies. This article will review this growing literature in an effort to evaluate the current clinical utility of endothelial dysfunction. PMID- 14522473 TI - Seven-year outcome in the RITA-2 trial: coronary angioplasty versus medical therapy. AB - OBJECTIVES: This study was designed to compare the long-term consequences of percutaneous transluminal coronary angioplasty (PTCA) and continued medical treatment. BACKGROUND: The long-term effects of percutaneous coronary intervention need evaluating, especially in comparison with an alternative policy of continued medical treatment. METHODS: The Second Randomized Intervention Treatment of Angina (RITA-2) is a randomized trial of PTCA versus conservative (medical) care in 1,018 patients considered suitable for either treatment option. Information on clinical events, interventions, and symptoms is available for a median seven years follow-up. RESULTS: Death or myocardial infarction (MI) occurred in 73 (14.5%) PTCA patients and 63 (12.3%) medical patients (difference +2.2%, 95% confidence interval -2.0% to +6.4%, p = 0.21). There were 43 deaths in both groups, of which 41% were cardiac-related. Among patients assigned PTCA 12.7% subsequently had coronary artery bypass grafts, and 14.5% required additional non-randomized PTCA. Most of these re-interventions occurred within a year of randomization, and after two years the re-intervention rate was 2.3% per annum. In the medical group, 35.4% required myocardial revascularization: 15.0% in the first year and an annual rate of 3.6% after two years. An initial policy of PTCA was associated with improved anginal symptoms and exercise times. These treatment differences narrowed over time, mainly because of coronary interventions in medical patients with severe symptoms. CONCLUSIONS: In RITA-2 an initial strategy of PTCA did not influence the risk of death or MI, but it improved angina and exercise tolerance. Patients considered suitable for PTCA or medical therapy can be safely managed with continued medical therapy, but percutaneous intervention is appropriate if symptoms are not controlled. PMID- 14522474 TI - Dilate or defer? View of a skeptic. PMID- 14522475 TI - The incidence, predictors, and outcomes of early reinfarction after primary angioplasty for acute myocardial infarction. AB - OBJECTIVES: We sought to identify the incidence, predictors, and clinical consequences of one-month reinfarction (RE-MI) in patients undergoing primary percutaneous coronary intervention (PCI) for acute myocardial infarction (AMI). BACKGROUND: One-month reinfarction after AMI significantly increases long-term mortality; however, little is known about the incidence and predictors of RE-MI in patients undergoing primary angioplasty. METHODS: We analyzed data from 3,646 patients who underwent primary PCI in the Primary Angioplasty in Acute Myocardial Infarction (PAMI) studies. We studied the incidence, correlates, and clinical outcomes of 30-day RE-MI. RESULTS: Reinfarction within one month of index hospitalization occurred in 77 (2.1%) of patients. In multivariate analysis, admission Killip class >1 (odds ratio [OR] 2.02, 95% confidence interval [CI] 1.09 to 3.76), left ventricular ejection fraction <50% (OR 2.49, 95% CI 1.30 to 4.74), final coronary stenosis >30% (OR 2.57, 95% CI 1.28 to 5.15), and presence of coronary dissection (OR 2.40, 95% CI 1.36 to 4.24) and thrombus (OR 2.36, 95% CI 1.23 to 4.53) on the final angiogram were independent correlates of RE-MI. One month reinfarction was independently associated with death (OR 7.14, 95% CI 3.28 to 15.5) and ischemic target vessel revascularization (I-TVR) (OR 15.0, 95% CI 8.68 to 26.0) at six months. CONCLUSIONS: We conclude that, although early RE-MI is uncommon in patients treated by primary PCI, it is a significant independent predictor of death and I-TVR at six months. Admission Killip class >1 and left ventricular systolic dysfunction were associated with higher incidence of RE-MI. Our results suggest that optimal revascularization during primary PCI may decrease RE-MI rates. PMID- 14522476 TI - Outcome of urgent and elective percutaneous coronary interventions after pharmacologic reperfusion with tenecteplase combined with unfractionated heparin, enoxaparin, or abciximab. AB - OBJECTIVES: The aim of this study was to evaluate percutaneous coronary intervention (PCI) in the Assessment of the Safety and Efficacy of New Thrombolytic Regimens (ASSENT-3) trial. BACKGROUND: In the ASSENT-3 trial, co therapy with abciximab (ABC) or enoxaparin (ENOX) reduced ischemic complications after ST-elevation acute myocardial infarction treated with tenecteplase when compared with unfractionated heparin (UFH). The effect of these new co-therapies on the results of PCI is unknown. METHODS: Clinical outcomes in patients who received co-therapy with ABC, ENOX, or UFH and subsequently underwent an elective (n = 1,064) or urgent (n = 716) PCI in the ASSENT-3 trial were compared. RESULTS: No significant differences in clinical end points were observed in patients who underwent an elective PCI. A non-significant trend toward fewer in-hospital myocardial re-infarctions was seen with ABC and ENOX when compared with UFH (0.5% vs. 0.6% vs. 1.5%, respectively). The incidence of bleeding complications was similar in the three treatment arms. Significantly fewer ABC- and ENOX-treated patients needed urgent PCI compared with UFH (9.1% vs. 11.9% vs. 14.3%; p < 0.0001), but outcomes in these patients were in general less favorable (30-day mortality: 8.2% vs. 5.4% vs. 4.5%; 1-year mortality: 11.0% vs. 8.5% vs. 5.6%; in hospital re-infarction: 3.9% vs. 2.5% vs. 2.7%; major bleeding complications: 8.8% vs. 7.0% vs. 3.4%). In pairwise comparisons with UFH, the higher one-year mortality and major bleeding rates after ABC were statistically significant (p = 0.045 and p = 0.012, respectively). CONCLUSIONS: Clinical outcomes after elective PCI were similar with the three antithrombotic co-therapies studied in ASSENT-3. Although fewer patients needed urgent PCI with ABC and ENOX, clinical outcomes were less favorable in this selected population, especially with ABC. PMID- 14522477 TI - Confusion in reperfusion. PMID- 14522478 TI - Triple antiplatelet therapy during percutaneous coronary intervention is associated with improved outcomes including one-year survival: results from the Do Tirofiban and ReoProGive Similar Efficacy Outcome Trial (TARGET). AB - OBJECTIVE: We sought to examine if clopidogrel treatment initiated before coronary stenting improved clinical outcomes among patients receiving aspirin and a glycoprotein (GP) IIb/IIIa inhibitor. BACKGROUND: Antiplatelet therapy plays a pivotal role in contemporary percutaneous coronary interventions (PCI). METHODS: Outcomes among 4,809 patients randomized to tirofiban or abciximab during PCI with stent placement were compared according to whether they received 300 mg of clopidogrel before PCI (93.1%) versus immediately after the procedure. RESULTS: The 30-day primary composite end point (death, myocardial infarction [MI], or urgent target vessel revascularization [TVR]) was lower among clopidogrel pretreated patients (6.6% vs. 10.4%, p = 0.009), mainly because of reduction of MI (6.0% vs. 9.5%, p = 0.012). The benefit of clopidogrel pretreatment was sustained at six months (death, MI, any TVR: 14.6% vs. 19.8%, HR = 0.71, p = 0.010), and this was due mainly to lowering of death and MI (7.8% vs. 13.0%, p = 0.001). At one year, clopidogrel pretreatment was associated with a lower mortality rate (1.7% vs. 3.6%, p = 0.011). Because clopidogrel pretreatment was not randomized, multivariable and propensity analyses were performed. After adjusting for baseline heterogeneity, clopidogrel pretreatment was an independent predictor for death or MI at 30 days (HR = 0.63, p = 0.012) and at six months (HR = 0.61, p = 0.003), and survival at one year (HR = 0.53, p = 0.044). No excess in 30-day bleeding events was noted with clopidogrel pretreatment. CONCLUSIONS: Among patients undergoing coronary stent placement with aspirin and a GP IIb/IIIa inhibitor, clopidogrel pretreatment is associated with a reduction of death and MI irrespective of the type of GP IIb/IIIa inhibitor used. PMID- 14522479 TI - Platelet inhibition strategies in percutaneous coronary intervention: competition or coopetition? PMID- 14522481 TI - The left atrium: a biomarker of chronic diastolic dysfunction and cardiovascular disease risk. PMID- 14522480 TI - Prediction of risk for first age-related cardiovascular events in an elderly population: the incremental value of echocardiography. AB - OBJECTIVES: We sought to determine if echocardiography enhances prediction of first age-related cardiovascular events. BACKGROUND: Whether echocardiographic assessment improves risk stratification for first cardiovascular events is not well known. METHODS: This retrospective cohort study included randomly selected residents of Olmsted County, Minnesota, age >/=65 years, who had >/=1 transthoracic echocardiograms at the Mayo Clinic between 1990 and 1998, in sinus rhythm, without valvular or congenital heart disease, and followed through medical records for first myocardial infarction (MI), coronary revascularization, atrial fibrillation (AF), congestive heart failure (CHF), transient ischemic attack (TIA), stroke, or cardiovascular death. Patients were excluded if they had any of these events before the baseline echocardiogram. RESULTS: Of 1,160 patients (age 75 +/- 7 years; 746 women) followed for a mean of 3.8 +/- 2.7 years, 333 (29%) first events occurred (70 AF, 67 coronary revascularization procedures, 65 CHF, 48 MI, 38 stroke, 25 TIA, and 20 cardiovascular deaths). In a multivariate model, age (p < 0.001), male gender (p < 0.001), diabetes mellitus (p = 0.005), systemic hypertension (p < 0.001), left atrial volume/body surface area >/=32 ml/m(2) (p = 0.003), left ventricular (LV) mass/height >/=120 g/m (p = 0.014), LV systolic dysfunction (p < 0.001), and LV diastolic dysfunction (p = 0.029) were independent predictors. A risk-scoring algorithm was developed and validated for the prediction of first events. The five-year event-free survival was 90%, 74%, and 50% for low-, medium-, and high-risk groups, respectively. CONCLUSIONS: Echocardiography enhanced prediction of first cardiovascular events in this referral-based elderly cohort. Its role in risk stratification for primary prevention of these events in the community warrants further investigations. PMID- 14522482 TI - Age does not limit quality of life improvement in cardiac valve surgery. AB - OBJECTIVES: We sought to determine the association of age with the change in quality of life (QOL) after valve surgery. BACKGROUND: Improvement in QOL is one of the principal goals of valve surgery. These procedures are being done with increasing frequency for older patients. METHODS: We prospectively studied 148 patients with aortic valve procedures and 72 patients with mitral valve procedures. Patients' QOL was measured at baseline and at 18 months using the Medical Outcomes Trust Short Form 36-Item (SF-36) Health Survey (response rate 90%). The association of age with change in QOL was measured by multiple regression analysis and based on two meta-scores of the SF-36: the Mental Component Summary (MCS) and the Physical Component Summary (PCS). RESULTS: Overall improvement in most domains of the SF-36, including the MCS and the PCS scores, was substantial. Improvement in the MCS score was not influenced by age in either aortic (0.09 score point improvement per 10-year age increments; p = 0.9) or mitral (0.90 score point improvement per 10-year age increments; p = 0.3) patients. Similarly, improvement in the PCS score did not vary by age in aortic patients (-1.00 score points per 10-year age increments; p = 0.2) and only slightly varied by age in mitral patients (-1.90 score points per 10-year age increments, p = 0.02). In the latter, despite statistical significance, the association was not substantial or clinically important. CONCLUSIONS: Among patients referred for cardiac valve surgery, age does not appear to limit the QOL benefits of surgery. PMID- 14522483 TI - Valve surgery in the elderly: question of quality (of life)? PMID- 14522484 TI - Is nutritional intake adequate in chronic heart failure patients? AB - OBJECTIVES: The goal of this study was to investigate the nutrition adequacy and energy availability for physical activity in free-living, clinically stable patients with chronic heart failure (CHF). BACKGROUND: Little information exists regarding the nutrition adequacy and alimentary habits of patients with clinically stable CHF. We hypothesized that CHF patients have an inadequate intake of calories and protein, leading to a negative calorie and nitrogen balance, an expression of increased tissue breakdown. METHODS: In 57 non-obese patients with CHF (52 males and 5 females; 52 +/- 3 years; body mass index <25 kg/m(2)) and in 49 healthy subjects (39 males and 10 females) matched for age, body mass index, and sedentary life style we evaluated total energy expenditure (TEE), calorie intake (kcal(I)), and nitrogen intake (N(I)) from a seven-day food diary, total nitrogen excretion (TNE), and energy availability (EA = kcal(I) - resting energy expenditure). A zero calorie balance (CB) occurred when kcal(I) = TEE; a nitrogen balance (NB) in equilibrium was set at NB (= N(I) - TNE) 0 +/- 1 g/day. RESULTS: In patients and controls kcal(I) and N(I) were similar. However, in CHF patients the kcal(I) was /=65 years with CHF identified through a 5% random sample of all U.S. Medicare beneficiaries. We assessed the relationship of the 20 most common noncardiac comorbidities to one year potentially preventable hospitalizations and total mortality. Preventable hospitalizations were determined by admissions for ambulatory care sensitive conditions using predefined criteria. RESULTS: Sixty-five percent of the sample had at least one hospitalization, of which 50% were potentially preventable. Exacerbations of CHF accounted for 55% of potentially preventable hospitalizations. Nearly 40% of patients with CHF had >/=5 noncardiac comorbidities, and this group accounted for 81% of the total inpatient hospital days experienced by all CHF patients. The risk of hospitalization and potentially preventable hospitalization strongly increased with the number of chronic conditions (both p < 0.0001). After controlling for demographic factors and other diagnoses, comorbidities that were associated consistently with notably higher risks for CHF-preventable and all-cause preventable hospitalizations, and mortality, included chronic obstructive pulmonary disease/bronchiectasis, renal failure, diabetes, depression, and other lower respiratory diseases (all p < 0.01). CONCLUSIONS: Noncardiac comorbidities are highly prevalent in older patients with CHF and strongly associate with adverse clinical outcomes. Cardiologists and other providers routinely caring for older patients with CHF may improve outcomes in this high-risk population by better recognizing non-CHF conditions, which may complicate traditional CHF management strategies. PMID- 14522487 TI - Delivering the cumulative benefits of triple therapy to improve outcomes in heart failure: too many cooks will spoil the broth. PMID- 14522488 TI - Hormone replacement therapy is associated with improved survival in women with advanced heart failure. AB - OBJECTIVES: We sought to determine whether hormone replacement therapy (HRT) is associated with an improved prognosis in women with advanced heart failure (HF) and systolic dysfunction. BACKGROUND: There are about two million postmenopausal women in the U.S. with HF. However, limited data are available to assess the effects of HRT on survival in this large group of patients. METHODS: A retrospective analysis of women age 50 years and over entered into the Beta Blocker Evaluation of Survival Trial (BEST) was conducted using Cox regression analysis comparing survival in HRT users and non-users after correcting for baseline variables known to predict survival in women with HF and systolic dysfunction. RESULTS: In 493 women age 50 years and older, HRT was associated with a significant reduction in mortality-21% mortality in HRT users and 34% in non-users (p = 0.025). Multivariate analysis demonstrated a hazard ratio for mortality of 0.6 (95% confidence interval = 0.36 to 0.97) (p = 0.039) for HRT users. The benefits of HRT were noted only in women with a nonischemic etiology of HF (n = 237). CONCLUSIONS: Hormone replacement therapy is associated with a marked improvement in survival in postmenopausal women with advanced HF. A prospective, randomized trial of HRT should be performed in this large group of patients. PMID- 14522489 TI - New hope for hormone replacement and the heart? PMID- 14522490 TI - Thoracoscopic extracardiac obliteration of the left atrial appendage for stroke risk reduction in atrial fibrillation. AB - OBJECTIVES: We evaluated left atrial appendage obliteration in high-risk patients with atrial fibrillation (AF). BACKGROUND: Left atrial appendage thrombosis and embolization is the principal mechanism of stroke in AF. Anticoagulation is underutilized and often contraindicated. METHODS: Thoracoscopic Left Appendage, Total Obliteration, No cardiac Invasion (LAPTONI) was undertaken with a loop snare in eight patients and a stapler in seven patients, median age 71 years, with clinical risk factors for stroke and with an absolute contraindication to or failure of prior thrombosis prevention with warfarin. Eleven patients had a history of prior thromboembolism. One patient took sustained warfarin during follow-up. RESULTS: The LAPTONI procedure was completed in 14 of 15 patients, and 1 patient required urgent conversion to open thoracotomy because of bleeding. Patients have been followed up for 8 to 60 months, mean 42 +/- 14 months. One fatal stroke occurred 55 months after surgery, and one non-disabling stroke three months after surgery. Two other deaths occurred, one after coronary bypass surgery and the other from hepatic failure. The subgroup of 11 patients with prior thromboembolism had an annualized rate of stroke of 5.2% per year (95% confidence interval [CI] 1.3 to 21) after LAPTONI, which compares to a rate of 13% per year (95% CI 9.0 to 19) for similar aspirin-treated patients from the Stroke Prevention in Atrial Fibrillation trials (p = 0.15). CONCLUSIONS: The LAPTONI procedure appears technically feasible without immediate disabling neurologic morbidity or mortality, and it demonstrates low post-operative event rates and a statistical trend toward thromboembolic risk reduction in high-risk AF patients. PMID- 14522491 TI - Role of left atrial appendage obliteration in stroke reduction in patients with mitral valve prosthesis: a transesophageal echocardiographic study. AB - OBJECTIVES: The aim of our study was to assess whether left atrial appendage (LAA) ligation in patients undergoing mitral valve replacement is associated with the risk of future embolisms. BACKGROUND: Previous studies show that the LAA plays an important role in the development of intracardiac thrombus. According to this decisive role, LAA surgical closure in patients undergoing cardiac surgery may be an attractive choice for reducing stroke. METHODS: We retrospectively studied 205 patients with previous mitral valve replacement and referred for echocardiography study. Patients were excluded if other causes of systemic embolism were found. The main outcome measure was the occurrence of an embolic event. RESULTS: Ligation of LAA was performed in 58 patients. However, an incomplete ligation was verified in six patients. During a median time from valve replacement to echocardiography study of 69.4 months (1 to 329), 27 patients had an embolism. Multivariate analysis identified the absence of LAA ligation (odds ratio [OR] 6.7 [95% confidence interval [CI] 1.5 to 31.0]; p = 0.02) and the presence of left atrial thrombus as the only independent predictors of occurrence of an embolic event. Moreover, when the identification of an incomplete LAA ligation was considered together with the absence of LAA ligation, risk of embolism increased up to 11.9 x (OR 11.9 [95% CI 1.5 to 93.6]; p = 0.02). CONCLUSIONS: Our study shows that LAA ligation during surgery of mitral valve replacement, performed in a high-risk population, is consistent with a reduction of the risk of late embolism and supports this technique if a mitral valve replacement is indicated. PMID- 14522492 TI - Obliteration of the left atrial appendage for prevention of thromboembolism. PMID- 14522493 TI - Competing autonomic mechanisms precede the onset of postoperative atrial fibrillation. AB - OBJECTIVES: This study was designed to evaluate autonomic changes preceding atrial fibrillation/flutter (AF) after thoracotomy. BACKGROUND: Autonomic fluctuations before the onset of postoperative AF have been reported but with conflicting results. METHODS: In 48 patients with postoperative AF, 2-h Holter recordings before the onset of AF were compared with corresponding data from 48 age- and gender-matched surgical controls without AF. Five-minute segments of heart rate variability (HRV) were studied using linear regression methods. RESULTS: There was a near-significant trend for the RR interval among patients with AF to be lower than controls (p = 0.06), whereas the standard deviation of RRs (p < 0.0001), root mean square of successive RR differences (p < 0.0001), proportion of RRs >50 ms different (p < 0.0001), low-frequency power (p = 0.0003) and its log (p < 0.0001), and high-frequency-power (p < 0.0001) and its log (p < 0.0001) were all significantly greater in patients with AF, respectively. In comparison to controls, AF patients had a significant decrease in RR interval (p = 0.02) and significant increments in all time- and frequency-domain analyses studied. CONCLUSIONS: In the period before the onset of postoperative AF, there are significant increases in HRV during a time when heart rate also increases. These novel findings are consistent with parasympathetic resurgence competing with increasing sympathetic activity as the triggering mechanism for postoperative AF. PMID- 14522494 TI - Autonomic modulation preceding the onset of atrial fibrillation. PMID- 14522495 TI - Catheter-induced linear lesions in the left atrium in patients with atrial fibrillation: an electroanatomic study. AB - OBJECTIVES: In this study using radiofrequency current and the electroanatomic mapping system CARTO, four line designs were tested in 84 patients suffering from drug-refractory atrial fibrillation (AFib). BACKGROUND: Prevention of AFib by trigger elimination within the pulmonary veins (PVs) has been recently reported, but the success may be lesser in patients with chronic AFib or large atria requiring linear lesion deployment. METHODS: Type A encircled the ostia of all four PVs with a connection to the mitral annulus (MA). In type B, three lines connected anatomic barriers. Type C encircled both septal and lateral PVs with connections between PVs and to the MA. Type D encircled PVs only. In the initial 12 patients (type D/1), line validation was performed without, and in 23 patients (type D/2) with, an additional catheter inside the encircled PVs. RESULTS: The ability to achieve completeness of all intended lines was 5% in type A, 21% in type B, 29% in C, 66% in type D/1, and 61% in type D/2. This resulted in stable sinus rhythm in 19% (4/21 patients) in type A, 32% (6/19 patients) in type B, 50% (7/14 patients) in type C, 58% (7/12 patients) in type D/1, and 65% (15/23 patients) in type D/2, respectively, over a mean follow-up of 620 +/- 376 days. Besides thromboembolic events (one stroke and one transient ischemic attack), total occlusion of a PV was a major complication in one patient, and acute tamponade in two patients. CONCLUSIONS: Complete lesions in the left atrium were difficult to achieve using conventional radiofrequency current technology, but were associated with sinus rhythm in 74% of patients during long-term follow-up, whereas incomplete lesions led mostly to recurrences of AFib or gap-related atrial tachycardia. PMID- 14522497 TI - Is there a referral bias against catheterization of patients with reduced left ventricular ejection fraction? Influence of ejection fraction and inducible ischemia on post-single-photon emission computed tomography management of patients without a history of coronary artery disease. AB - OBJECTIVES: The objective of this work was to define the relationship between left ventricular perfusion/ function measures and referral rates to catheterization and revascularization early after stress gated myocardial perfusion single-photon emission computed tomography (MPS). BACKGROUND: Although revascularization yields the greatest survival benefit in patients with low ejection fraction (EF) and extensive coronary artery disease, referral patterns to catheterization and revascularization after noninvasive testing are not well defined. METHODS: We identified 3,369 patients without previous myocardial infarction or revascularization who underwent exercise or adenosine stress MPS and who were followed-up (97% complete) for occurrence of early (<60 days) post single-photon emission computed tomography (SPECT) revascularization. Multivariable logistic regression modeling was used to determine the association of various patient characteristics and test results with performance of catheterization and revascularization as separate end points. RESULTS: In the first 60 days after stress MPS, 445 catheterizations (13.2%) and 254 revascularizations (7.5%) occurred, including 140 coronary artery bypass graft surgeries (4.1%) and 114 percutaneous coronary interventions (3.4%). Both post stress gated EF and percent of the myocardium ischemic by stress MPS were independent predictors of revascularization. Logistic regression revealed that the likelihood of catheterization increased with both increasing ischemia and decreasing EF (c-index = 0.94, chi-square = 590). Predicted referral rates to catheterization increased with decreasing EF except in patients with severe ischemia (>15% of myocardium), where rates decreased with decreasing EF. Similar modeling of revascularization (c-index = 0.94, chi-square = 329) revealed that the likelihood of revascularization increased with increasing ischemia but, in general, decreased with decreasing EF. CONCLUSIONS: Although post-SPECT referral to both catheterization and revascularization is driven by ischemia, EF has the opposite effect on these two outcomes. Further studies evaluating the appropriateness of these referral patterns are warranted. PMID- 14522496 TI - Linear ablation for atrial fibrillation: have we come full circle?. PMID- 14522498 TI - Clinical safety of magnetic resonance imaging early after coronary artery stent placement. AB - OBJECTIVES: Our aim was to examine the rate of adverse cardiac events in patients undergoing magnetic resonance imaging (MRI) <8 weeks after coronary stent placement. BACKGROUND: The risk of coronary stent thrombosis from dislodgement due to MRI early after stent placement is not well defined. Manufacturers recommend postponing MRI studies until eight weeks after coronary stent placement. METHODS: We analyzed the Mayo Clinic Rochester Percutaneous Coronary Intervention Database and examined records of 111 patients who underwent MRI <8 weeks after coronary stent placement treated with aspirin and a thienopyridine. Occurrence of death, myocardial infarction (MI), and repeat revascularization within 30 days of MRI were recorded. RESULTS: Magnetic resonance imaging (1.5 tesla) was performed within a median of 18 days (range, 0 to 54 days) after coronary stent placement. Four noncardiac deaths occurred, and three patients had repeat revascularization procedures. Stent thrombosis did not occur (95% confidence interval, 0% to 3.3%). CONCLUSIONS: Magnetic resonance imaging <8 weeks after coronary stent placement appears to be safe, and the risk of cardiac death or MI due to stent thrombosis is low. Postponing MRI does not appear to be necessary. PMID- 14522499 TI - Effect of tumor necrosis factor-alpha on endothelial and inducible nitric oxide synthase messenger ribonucleic acid expression and nitric oxide synthesis in ischemic and nonischemic isolated rat heart. AB - OBJECTIVES: The present study aimed to investigate the influence of endogenous tumor necrosis factor-alpha (TNF-alpha) that was synthesized during ischemia and exogenous TNF-alpha on endothelial and inducible nitric oxide synthase (eNOS and iNOS) messenger ribonucleic acid (mRNA) expression and nitric oxide (NO) production in the isolated rat heart. BACKGROUND: Tumor necrosis factor-alpha is recognized as being a proinflammatory cytokine with a significant cardiodepressant effect. One of the proposed mechanisms for TNF-alpha-induced cardiac contractile dysfunction is increased NO production via iNOS mRNA upregulation, but the role of NO in TNF-alpha-induced myocardial dysfunction is highly controversial. METHODS: Isolated rat hearts studied by a modified Langendorff model were randomly divided into subgroups to investigate the effect of 1-h global cardioplegic ischemia or the effect of 1-h perfusion with exogenous TNF-alpha on the expression of eNOS mRNA and iNOS mRNA and on NO production. RESULTS: After 1 h of ischemia, there were significant increases in TNF levels in the effluent (from hearts), and eNOS mRNA expression had declined (from 0.91 +/- 0.08 to 0.68 +/- 0.19, p < 0.001); but there were no changes in iNOS mRNA expression, and NO was below detectable levels. Perfusion of isolated hearts with TNF-alpha had a cardiodepressant effect and decreased eNOS mRNA expression to 0.67 +/- 0.04 (p < 0.002). Inducible nitric oxide synthase mRNA was unchanged, and NO was below detectable levels. CONCLUSIONS: We believe this is the first study to directly show that TNF-alpha does not increase NO synthesis and release but does downregulate eNOS mRNA in the ischemic and nonischemic isolated rat heart. PMID- 14522500 TI - Androgens up-regulate atherosclerosis-related genes in macrophages from males but not females: molecular insights into gender differences in atherosclerosis. AB - OBJECTIVES: This study investigated the effects of androgens on gene expression in male- and female-donor macrophages. BACKGROUND: Men have more severe coronary disease than women. Androgen exposure increases foam cell formation in male but not female macrophages, and male macrophages express >4-fold more androgen receptor messenger ribonucleic acid than female macrophages. Therefore, androgen exposure may have gender-specific and potentially pro-atherogenic effects in macrophages. METHODS: Utilizing complementary deoxyribonucleic acid arrays, we studied the effects of a pure androgen (dihydrotestosterone, 40 nmol/l) on human monocyte-derived macrophages from healthy male and female donors (n = 4 hybridizations; 2 men, 2 women). Differential expression of atherosclerosis related genes was confirmed by real-time reverse transcription-polymerase chain reaction (RT-PCR) in five male and five female donors. Functional corroboration of foam cell formation-related findings was undertaken by experiments using (125)I-acetylated low-density lipoprotein (AcLDL). RESULTS: In male macrophages, androgen treatment produced differential up-regulation of 27 genes concentrated in five functional classes: 1) lipoprotein processing; 2) cell-surface adhesion; 3) extracellular signaling; 4) coagulation and fibrinolysis; and 5) transport protein genes. By contrast, none of 588 genes were up-regulated in female macrophages. By RT-PCR, we confirmed the gender-specific up-regulation of six of these atherosclerosis-related genes: acyl coenzyme A:cholesterol acyl transferase I, lysosomal acid lipase (LAL), caveolin-2, CD40, vascular endothelial growth factor-165 receptor, and tissue factor pathway inhibitor. Functionally, androgen treated male macrophages showed increased rates of lysosomal AcLDL degradation, by 45% to 75% after 15 to 20 h of (125)I-AcLDL incubation (p = 0.001), consistent with increased LAL activity. CONCLUSIONS: Androgens increase expression of atherosclerosis-related genes in male but not female macrophages, with functional consequences. These findings may contribute to the male predisposition to atherosclerosis. PMID- 14522501 TI - What constitutes a great review? PMID- 14522502 TI - Rethink retirement: plan a second career in cardiology. PMID- 14522504 TI - Is the tolerability of long-term thiazolidinedione therapy overstated? PMID- 14522503 TI - ACC/AHA/ASNC guidelines for the clinical use of cardiac radionuclide imaging- executive summary: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines (ACC/AHA/ASNC Committee to Revise the 1995 Guidelines for the Clinical Use of Cardiac Radionuclide Imaging). PMID- 14522506 TI - Is diastolic heart failure synonymous with heart failure with preserved ejection fraction? PMID- 14522508 TI - Determination of the natural history of aspirin resistance among stable patients with cardiovascular disease. PMID- 14522512 TI - Listeria monocytogenes infection in patients with cancer. AB - Listeriosis (LT) is an important infection in immunocompromised patients, but no large series of LT in cancer patients have been recently described. We reviewed the records of 34 cancer patients with LT at our institution (1990-2001). Twenty patients (59%) had an underlying hematologic malignancy. In 11 patients, LT complicated bone marrow transplantation. Lymphocytopenia was observed in 62% of the patients. Twenty-six patients (76%) received prior corticosteroids. Bacteremia was the most common presentation of LT (74%) followed by meningoencephalitis (21%). The most common treatment of LT was ampicillin with or without gentamicin (68%). The median duration of treatment was 26 days (range, 8 74 days). The rate of response to antimicrobial therapy was 79%. No relapses were identified. LT contributed to death in 9 (75%) of the 12 patients who died. Meningoencephalitis had the worst prognosis (3 of 6 cases were fatal). Treatment of central nervous system LT continues to have a high failure rate. PMID- 14522511 TI - Ribotyping characterisation of campylobacter isolates randomly collected from different sources in Italy. AB - In this study the potential for using the automated PstI ribotyping as a primary library typing method to survey Campylobacter and for identification of two thermophilic Campylobacter jejuni and Campylobacter coli species was evaluated. A total of 158 isolates randomly collected in Italy from different sources were analyzed. A large percentage of chicken (28%), turkey (27%) and turkey meat (25%) isolates shared their ribotyping profiles (ribotypes) with those of humans, whereas the swine isolates had unique profiles. The identification results obtained by ribotyping corresponded to those collected by using a multiplex PCR protocol specifically designed for C. jejuni and C. coli detection. The comparison of the PstI ribotyping profiles obtained in this research with those of the isolates collected over time will facilitate determining the ribotypes that are more frequently transmitted to humans in comparison to those that are normally harboured only in animals, foods and in the environment. PMID- 14522513 TI - Prevalence of enterotoxigenic Escherichia coli (ETEC) in hospitalized acute diarrhea patients in Denpasar, Bali, Indonesia. AB - The relationship between enterotoxigenic Escherichia coli (ETEC) and hospitalized patients with acute diarrhea was examined in a study conducted in two hospitals from June 2000 to May 2001 in Denpasar, Bali, Indonesia. A total of 489 hospitalized patients with acute diarrhea were enrolled, and their rectal swabs were screened for enteric bacterial pathogens. Toxins, colonization factor antigens (CFAs), in vitro antimicrobial susceptibility and seasonal distribution patterns associated with ETEC were ascertained. The diagnosis of ETEC infection and CFAs association were performed with GM-1 ELISA and Dot blot immunoassays. Enterotoxigenic Escherichia coli was isolated from the rectal swabs of 14.9% of the patients. The distribution of toxins among the ETEC strains found was ST in 51 (69.9%), while LT and ST/LT were found in 28.8% and 1.3% respectively. The highest isolation rate for ETEC was found among children between the ages of 1 and 15 years. Colonization factor antigens were identified in 28.8% of the ETEC strains. A high prevalence of CFA was found among the rectal swabs of patients with ST isolates. High frequency of resistance to ampicillin, trimethoprim/sulfamethoxazole, chloramphenicol, tetracycline and cephalothin was displayed among the ETEC strains. All ETEC strains were susceptible to norfloxacin, ciprofloxacin and nalidixic acid. The results of this study document the prevalence of ETEC in hospitalized patients with acute diarrhea in Denpasar, Bali, Indonesia. Data generated in this study depicts the prevalence of ETEC diarrhea and CFA types among diarrhea patients in the tourist city of Denpasar, Bali, Indonesia. PMID- 14522514 TI - A quantitative LightCycler PCR to detect Streptococcus pneumoniae in blood and CSF. AB - A quantitative real-time PCR targeting the Pneumolysin (ply) gene of Streptococcus pneumoniae was developed for the LightCycler instrument using Fluorescence Resonance Energy Transfer (FRET) probes. All common S. pneumoniae serotypes were detected while other bacteria and viruses were not. The sensitivity was determined to be between one and ten target copies per reaction. The PCR was applied to six CSF and 16 whole blood specimens from 17 patients with laboratory proven invasive pneumococcal disease. One hundred percent of CSF specimens and 69% of whole blood specimens were PCR positive. The bacterial loads were determined to be 7.6 to 6.01 x 10(5) copies/microL for the six CSF specimens, and 0.08 to 5.4 x 10(2) copies/microL for the 16 whole blood specimens. Ninety-seven percent of 30 culture and Gram's stain negative CSF specimens and 100% of 50 normal whole blood specimens were PCR negative. This highly sensitive and specific PCR assay has the potential to provide sufficiently rapid results to improve antibiotic treatment of S.pneumoniae infections, while bacterial load quantitation has opened up exciting possibilities for patient management. PMID- 14522515 TI - Sequential exposure of Aspergillus fumigatus to itraconazole and caspofungin: evidence of enhanced in vitro activity. AB - We investigated the in vitro activity of sequential itraconazole and caspofungin against 10 isolates of Aspergillus fumigatus. Previous exposure of A. fumigatus to itraconazole resulted in dose-dependent enhanced effects of caspofungin and vice versa. Our finding suggests a preferential role for azole-caspofungin sequential combinations and merits further in vivo investigation. PMID- 14522516 TI - Routine use of a simple low-cost genotypic assay for the identification of mycobacteria in a high throughput laboratory. AB - A novel polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis (PRA) of the hsp65 gene was used for the routine identification of mycobacteria in a high throughput clinical laboratory. A total of 2036 clinical isolates were tested by PRA in conjunction with other methods. The PRA identification of M. tuberculosis complex was 100% sensitive and specific, and 74.5% of nontuberculous mycobacteria (NTM) were correctly identified. It gave highly consistent results for Mycobacterium avium complex (MAC) species and for most isolates of M. fortuitum, M. chelonae, and M. kansasii. It had proven to be highly robust and stable despite usage on such a large-scale and is thus particularly suitable for use in high throughput laboratories in areas with a high incidence of tuberculosis. PMID- 14522517 TI - Left-sided endocarditis caused by Pseudomonas aeruginosa: successful treatment with meropenem and tobramycin. AB - Medical treatment alone is rarely successful in left-sided infective endocarditis caused by Pseudomonas aeruginosa. We report the cure of such a case with high dose meropenem in combination with tobramycin. PMID- 14522518 TI - Pericarditis as primary manifestation of Mycobacterium bovis SSP. caprae infection. AB - A 76-year-old white male presented with progressive malaise, weight loss and dyspnea at rest. Echocardiography revealed a circular pericardial effusion and global hypokinesia. Pericardiocentesis showed a purulent exudate and microbiologic examination revealed Mycobacterium bovis fully sensitive to isoniazid, streptomycin, ethambutol, rifampin, and pyrazinamide. By spoligotyping the isolate could be further differentiated to M. bovis ssp. caprae. Antimycobacterial therapy was initiated but 3 weeks later the patient's circulation and renal function deteriorated and he died with clinical signs of sepsis despite intensive care treatment. Pericarditis is a rare manifestation of tuberculosis and can be fatal even when diagnosed and treated appropriately. In low incidence countries diagnosis is often delayed and even overlooked. PMID- 14522519 TI - Comparative activity and spectrum of broad-spectrum beta-lactams (cefepime, ceftazidime, ceftriaxone, piperacillin/tazobactam) tested against 12,295 staphylococci and streptococci: report from the SENTRY antimicrobial surveillance program (North America: 2001-2002). AB - A contemporary collection of 12,295 North American isolates (2001-2002) consisting of Staphylococcus aureus (50%), coagulase-negative staphylococci (12%), Streptococcus pneumoniae (24%), beta-hemolytic streptococci (12%), and viridans-group streptococci (2%) were tested against broad-spectrum beta-lactams (cefepime, ceftazidime, ceftriaxone, imipenem, piperacillin/tazobactam) and comparator agents using a reference broth microdilution method to determine their continued effectiveness for empiric antimicrobial therapy. All isolates were very susceptible to vancomycin, linezolid and quinupristin/dalfopristin (>98%). Oxacillin-susceptible staphylococci were also highly susceptible to the tested beta-lactams (>98%) with the exception of ceftazidime (93%). beta-hemolytic streptococci were exquisitely susceptible (>99%) to penicillin and all other agents except for clindamycin (94%) and erythromycin (81%). Viridans group streptococci were routinely less susceptible than were other streptococci. S. pneumoniae remained susceptible to most agents (>91%) with the exceptions of erythromycin (74%) and penicillin (69%). Among beta-lactams tested against S. pneumoniae, ceftriaxone and cefepime continued to be very active against penicillin-susceptible (>99%) and intermediate (>98%) strains, but less active (80% and 82%, respectively) against penicillin-resistant isolates. These findings confirm that the newer cephalosporins (cefepime and ceftriaxone) among broad spectrum beta-lactam agents have a spectrum of activity that remains comprehensive for the commonly isolated Gram-positive pathogens. PMID- 14522520 TI - In vitro antimicrobial activity of moxifloxacin compared to other quinolones against recent clinical bacterial isolates from hospitalized and community-based cancer patients. AB - The in vitro spectrum of moxifloxacin (a C-8-methoxyquinolone) was compared to that of ciprofloxacin and levofloxacin against 924 recent clinical isolates from cancer patients. Moxifloxacin was more active than the comparator agents against Gram-positive pathogens, with potent activity against Aerococcus spp., Listeria monocytogenes, Micrococcus spp., Rhodococcus equi, and Stomatococcus mucilaginous, methicillin-susceptible Staphylococcus spp., all beta hemolytic streptococci, viridans streptococci and Streptococcus pneumoniae. It also had good to moderate activity against Bacillus spp., Corynebacterium spp., Enterococcus faecalis, and methicillin-resistant staphylococci. Although ciprofloxacin was the most active agent tested against the Enterobacteriaceae, moxifloxacin inhibited the majority of these isolates at < or =2.0 microg/ml. Moxifloxacin was the least active of the three agents tested against Pseudomonas aeruginosa, but had significant activity against other non-fermentative Gram negative bacilli including Acinetobacter spp., Flavobacterium spp., Pseudomonas spp. other than P. aeruginosa, and Stenotrophomonas maltophilia. The overall broad spectrum of moxifloxacin, and its availability for both oral and parenteral administration, warrants its evaluation for the prevention and treatment of infections in cancer patients. PMID- 14522521 TI - Comment on "First report of a catalase-negative methicillin-resistant Staphylococcus aureus". PMID- 14522524 TI - One standard, not two. PMID- 14522525 TI - Selective digestive decontamination: for everyone, everywhere? PMID- 14522526 TI - Ischaemic tolerance: a window to endogenous neuroprotection? PMID- 14522527 TI - HIV-1, antiretroviral therapy, and malaria. PMID- 14522528 TI - Registration of trials and protocols. PMID- 14522530 TI - Effects of selective decontamination of digestive tract on mortality and acquisition of resistant bacteria in intensive care: a randomised controlled trial. AB - BACKGROUND: Selective decontamination of the digestive tract (SDD) is an infection-prevention regimen used in critically ill patients. We assessed the effects of SDD on intensive-care-unit (ICU) and hospital mortality, and on the acquisition of resistant bacteria in adult patients admitted to intensive care. METHODS: We did a prospective, controlled, randomised, unblinded clinical trial. 934 patients admitted to a surgical and medical ICU were randomly assigned oral and enteral polymyxin E, tobramycin, and amphotericin B combined with an initial 4-day course of intravenous cefotaxime (SDD group n=466), or standard treatment (controls n=468). Primary endpoints were ICU and hospital mortality and the acquisition of resistant bacteria. FINDINGS: In the SDD group 69 (15%) patients died in the ICU compared with 107 (23%) in the control group (p=0.002). Hospital mortality was lower in the SDD groups than in the control group (113 [24%] vs 146 [31%], p=0.02). During their stay in intensive care, colonisation with gram negative bacteria resistant to ceftazidime, ciprofloxacin, imipenem, polymyxin E, or tobramycin occurred in 61 (16%) of 378 SDD patients and in 104 (26%) of 395 patients in the control group (p=0.001). Colonisation with vancomycin-resistant enterococcus occurred in five (1%) SDD patients and in four (1%) controls (p=1.0). No patient in either group was colonised with meticillin-resistant Staphylococcus aureus. INTERPRETATION: In a setting with low prevalence of vancomycin-resistant enterococcus and meticillin-resistant S aureus, SDD can decrease ICU and hospital mortality and colonisation with resistant gram-negative aerobic bacteria. PMID- 14522531 TI - Formation and early development of follicles in the polycystic ovary. AB - BACKGROUND: Polycystic ovary syndrome is the most common cause of anovulatory infertility. It has long-term health implications and is an important risk factor for type 2 diabetes. However, little is known about the cause of polycystic ovaries. We have used detailed morphological analysis to assess the hypothesis that there is an intrinsic ovarian abnormality that affects the earliest stages of follicular development. METHODS: We took small cortical biopsies during routine laparoscopy from 24 women with normal ovaries and regular cycles and from 32 women with polycystic ovaries, 16 of whom had regular, ovulatory cycles and 16 of whom had oligomenorrhoea. We used computerised image analysis to assess the density and developmental stage of small preantral follicles in serial sections of fixed tissue. FINDINGS: Median density of small preantral follicles, including those at primordial and primary stages, was six-fold greater in biopsies from polycystic ovaries in anovulatory women than in normal ovaries (p=0.009). In both ovulatory and anovulatory women with polycystic ovaries, we noted a significant increase in the percentage of early growing (primary) follicles and a reciprocal decrease in the proportion of primordial follicles compared with normal ovaries. INTERPRETATION: Our findings indicate that there are fundamental differences between polycystic and normal ovaries in early follicular development, suggesting an intrinsic ovarian abnormality. The increased density of small preantral follicles in polycystic ovaries could result from increased population of the fetal ovary by germ cells, or from decreased rate of loss of oocytes during late gestation, childhood, and puberty. PMID- 14522532 TI - Validity of reported vaccination coverage in 45 countries. AB - BACKGROUND: Monitoring and assessment of coverage rates in national health programmes is becoming increasingly important. We aimed to assess the accuracy of officially reported coverage rates of vaccination with diphtheria-tetanus pertussis vaccine (DTP3), which is commonly used to monitor child health interventions. METHODS: We compared officially reported national data for DTP3 coverage with those from the household Demographic and Health Surveys (DHS) in 45 countries between 1990 and 2000. We adjusted survey data to reflect the number of valid vaccinations (ie, those administered in accordance with the schedule recommended by WHO) using a probit model with sample selection. The model predicted the probability of valid vaccinations for children, including those without documented vaccinations, after correcting for bias from differences between the children with and without documented information on vaccination. We then assessed the extent of survey bias and differences between officially reported data and those from DHS estimates. FINDINGS: Our results suggest that officially reported DTP3 coverage is higher than that reported from household surveys. This size of the difference increases with the rate of reported coverage of DTP3. Results of time-trend analysis show that changes in reported coverage are not correlated with changes reported from household surveys. INTERPRETATION: Although reported data might be the most widely available information for assessment of vaccination coverage, their validity for measuring changes in coverage over time is highly questionable. Household surveys can be used to validate data collected by service providers. Strategies for measurement of the coverage of all health interventions should be grounded in careful assessments of the validity of data derived from various sources. PMID- 14522534 TI - A man with fever and a persistent handgrip. PMID- 14522533 TI - Effect of ischaemic preconditioning on genomic response to cerebral ischaemia: similarity to neuroprotective strategies in hibernation and hypoxia-tolerant states. AB - BACKGROUND: Molecular mechanisms of neuroprotection that lead to ischaemic tolerance are incompletely understood. Identification of genes involved in the process would provide insight into cell survival and therapeutic approaches for stroke. We developed a mouse model of neuroprotection in stroke and did gene expression profiling to identify potential neuroprotective genes and their associated pathways. METHODS: Eight mice per condition were subjected to occlusion of the middle cerebral artery for 15 min (preconditioning), 60 min (injurious ischaemia), or preconditioning followed 72 h later by injurious ischaemia. RNA was extracted from the cortical regions of the ischaemic and non ischaemic hemispheres. Three pools per condition were generated, and RNA was hybridised to oligonucleotide microarrays for comparison of ischaemic and non ischaemic hemispheres. Real-time PCR and western blots were used to validate results. Follow-up experiments were done to address the biological relevance of findings. FINDINGS: Microarray analysis revealed changes in gene expression with little overlap among the conditions of injurious ischaemia, ischaemic preconditioning, or both. Injurious ischaemia induced upregulation of gene expression; 49 (86%) of 57 genes regulated showed increased expression in the ischaemic hemisphere. By contrast, preconditioning followed by injurious ischaemia resulted in pronounced downregulation; 47 (77%) of 61 regulated genes showed lower expression. Preconditioning resulted in transcriptional changes involved in suppression of metabolic pathways and immune responses, reduction of ion-channel activity, and decreased blood coagulation. INTERPRETATION: Preconditioning reprogrammes the response to ischaemic injury. Similar changes reported by others support an evolutionarily conserved endogenous response to decreased blood flow and oxygen limitation such as seen during hibernation. PMID- 14522535 TI - Effect of HIV-1 antiretroviral drugs on cytoadherence and phagocytic clearance of Plasmodium falciparum-parasitised erythrocytes. AB - In response to the HIV/AIDS pandemic, antiretroviral treatment is currently being implemented in sub-Saharan Africa, where malaria and HIV-1 co-infections are highly prevalent. The effectiveness and tolerability of antiretrovirals in patients with malaria and HIV-1 co-infection have not been investigated. Antiretrovirals decrease CD36 surface concentrations in vivo, which might impair receptor function and affect parasite-host interactions. Thus, we investigated the effects of these drugs on CD36-mediated cytoadherence and non-opsonic phagocytosis of Plasmodium falciparum parasitised erythrocytes in vitro. The protease-inhibitor class of antiretrovirals particularly impairs CD36-mediated cytoadherence and non-opsonic phagocytosis of parasitised erythrocytes by human macrophages. Such treatment might therefore contribute to altered malaria disease outcomes in co-infected patients. PMID- 14522536 TI - Deaths due to absence of an affordable antitoxin for plant poisoning. AB - There is a severe shortage of affordable antivenoms and antitoxins in the developing world. An anti-digoxin antitoxin for oleander poisoning was introduced in Sri Lanka in July, 2001, but because of its cost, stocks ran out in July, 2002. We looked at the effect of its introduction and withdrawal on case fatality, and determined its cost-effectiveness. The antitoxin strikingly reduced the case fatality; its absence resulted in a three-fold rise in deaths. At the present price of US2650 dollars per course, every life saved cost 10209 dollars and every life year cost 248 dollars. Reduction of the antitoxin's price to 400 dollars would reduce costs to 1137 dollars per life gained; a further reduction to 103 dollars would save money for every life gained. Treatments for poisoning and envenoming should be included in the present campaign to increase availability of affordable treatments in the developing world. PMID- 14522537 TI - Chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a major cause of death and disability worldwide. Recognition that the burden of this disorder will continue to increase over the next 20 years despite medical intervention has stimulated new research into the underlying mechanisms, leading to a rational basis for evaluation of existing therapies, and has suggested novel treatment approaches. Tobacco exposure remains the main but not exclusive cause of COPD. Whether the lung is injured by changes in the balance of proteases and antiproteases, tissue damage by oxidative stress, or a combination of the two is still not known. The genetic basis of susceptibility to COPD is now being studied as is the role of computed tomography in the identification of structural damage in individuals with less symptomatic disease. Clinical diagnosis still relies heavily on an appropriate history confirmed by abnormal spirometry. Smoking cessation is possible in a substantial proportion of individuals with symptoms but is most effective if withdrawal is supported by pharmacological treatment. Treatment with long-acting inhaled bronchodilators and, in more severe disease, inhaled corticosteroids reduces symptoms and exacerbation frequency and improves health status. Rehabilitation can be even more effective, at least for a year after the treatment. Recent guidelines have made practical suggestions about how to optimise these treatments and when to consider addition of oxygen, surgery, and non-invasive ventilation. Regular review of this guidance is important if future management advances are to be implemented effectively. PMID- 14522539 TI - Hepatitis A booster vaccination: is there a need? AB - Hepatitis A is one of the most common vaccine-preventable infectious diseases in the world. Effective vaccines against hepatitis A have been available since 1992, and they provide long-term immunity against the infection. However, there is no worldwide consensus on how long protection will last or whether there will be a need for hepatitis A virus (HAV) booster vaccinations in the future. In most countries, booster-vaccination policy is guided by manufacturers' recommendations, national authorities, or both. In June, 2002, a panel of international experts met to review the long-term immunogenicity and protection conferred by HAV vaccine in different population groups. Data have shown that after a full primary vaccination course, protective antibody amounts persist beyond 10 years in healthy individuals, and underlying immune memory provides protection far beyond the duration of anti-HAV antibodies. The group concluded that there is no evidence to lend support to HAV booster vaccination after a full primary vaccination course in a healthy individual. However, further investigations are needed before deciding if boosters can be omitted in special patient-groups. PMID- 14522538 TI - Mycolactones and Mycobacterium ulcerans disease. AB - CONTEXT: Mycobacterium ulcerans causes devastating necrotic lesions in affected individuals. The disease, commonly called Buruli ulcer, is increasing in prevalance in western African countries. Treatment is mainly surgical; no clinical trials have been done to support the use of antimycobacterial drugs. A secreted polyketide toxin, mycolactone, is responsible for the tissue damage; its chemical structure has been elucidated. STARTING POINT: Although the main treatment is surgical, many patients with Buruli ulcer present late because of unusual beliefs about the disease and its treatment. Isabelle Aujoulat and colleagues recently showed, in a study in southern Benin, Africa (Trop Med Int Health 2003; 8: 750-59), that although the ulcer is well recognised, the cause is often seen as environmental or because of witchcraft. In addition, treatment is thought to be destructive, costly, and ineffective. WHERE NEXT? Antimycobacterial drug regimens that hold promise based on animal and preliminary human studies will soon be tested in large well-designed controlled clinical trials. Information gleaned from the genomic sequence of M ulcerans could be used to design more effective vaccines, or new drug targets (eg, that knock out the enzymes of M ulcerans that synthesise mycolactone species). PMID- 14522540 TI - The public health implications of world trade negotiations on the general agreement on trade in services and public services. AB - Trade ministries from the World Trade Organization's (WTO's) 144 member states are presently deciding which public services to open to foreign competition under the complex liberalisation rules of the general agreement on trade in services (GATS). A frequent criticism of the WTO system is that it reduces national autonomy over public policy. However, respect for national sovereignty is asserted in the GATS treaty. Here, we examine claims made by the WTO and others that GATS exempts public services and does not require their privatisation. We discuss trade treaty processes that can subject public services to commercial rules, the treaty's flexibility with respect to national autonomy, and the effect of GATS in situations in which national autonomy is not protected. We conclude that national autonomy over health policy is not preserved under GATS, and that accordingly, there is a role for international standards that protect public services from the adverse effect of trade and market forces. PMID- 14522541 TI - COMET: a proposed mechanism of action to explain the results and concerns about dose. PMID- 14522542 TI - COMET: a proposed mechanism of action to explain the results and concerns about dose. PMID- 14522544 TI - COMET: a proposed mechanism of action to explain the results and concerns about dose. PMID- 14522545 TI - Confidentiality. PMID- 14522546 TI - Addressing inequities in global health. PMID- 14522547 TI - Nebulised salbutamol and magnesium sulphate in acute asthma. PMID- 14522548 TI - Nebulised salbutamol and magnesium sulphate in acute asthma. PMID- 14522549 TI - Nebulised salbutamol and magnesium sulphate in acute asthma. PMID- 14522551 TI - Effect of conjugate pneumococcal vaccine on recurrent acute otitis media. PMID- 14522553 TI - Road medicine and HIV/AIDS. PMID- 14522555 TI - Palliative chemotherapy: a clinical oxymoron. PMID- 14522556 TI - The role of the surgeon in translational research. PMID- 14522557 TI - Programme evaluation in CME. PMID- 14522560 TI - The curse of awareness. PMID- 14522561 TI - It's not the winning... PMID- 14522562 TI - The importance of cholesterol, blood pressure and smoking for coronary heart disease. PMID- 14522563 TI - Neurohormonal inhibition in heart failure, is there no limit? PMID- 14522564 TI - Drug-drug interactions involving antiplatelet agents. PMID- 14522565 TI - Clinical applications of B-type natriuretic peptide (BNP) testing. AB - Many claims have been made in recent years regarding the utility of plasma B-type natriuretic peptide (BNP) concentration measurements in the diagnosis, risk stratification and monitoring of patients with heart failure. This paper summarizes the current evidence and provides guidance for practising clinicians. Overall, plasma BNP testing appears to be of most value in the diagnostic arena, where it is likely to improve the performance of non-specialist physicians in diagnosing heart failure. In clinical practice, BNP testing is best used as a 'rule out' test for suspected cases of new heart failure in breathless patients presenting to either the outpatient or emergency care settings; it is not a replacement for echocardiography and full cardiological assessment, which will be required for patients with an elevated BNP concentration. Although work is ongoing in establishing the 'normal' values of BNP, heart failure appears to be highly unlikely below a plasma concentration of 100 pg/ml. However, as BNP levels rise with age and are affected by gender, comorbidity and drug therapy, the plasma BNP measurement should not be used in isolation from the clinical context. PMID- 14522566 TI - Re-assessing the contribution of serum total cholesterol, blood pressure and cigarette smoking to the aetiology of coronary heart disease: impact of regression dilution bias. AB - AIMS: To estimate the combined contribution of serum total cholesterol, blood pressure and cigarette smoking to coronary heart disease (CHD) risk after adjustment for regression dilution bias. METHODS AND RESULTS: Six thousand, five hundred and thirteen middle-aged British men without CHD were followed for major CHD events over 10 years. The population attributable risk fraction (PARF) was predicted for a range of risk factor thresholds before and after adjustment for regression dilution of serum total cholesterol and blood pressure. Defining 'low risk' individuals as being in the bottom tenth of the population distributions of serum total cholesterol (<5.2 mmol/l) and diastolic blood pressure (<70 mmHg) and a non-cigarette smoker, the PARF was 75%, increasing to 86% after adjustment for regression dilution. Regardless of the threshold criteria chosen, the PARF was substantially greater than 65% before adjustment for regression dilution and greater than 75% after adjustment. Exclusion of ex-smokers and passive smokers from the low-risk group increased estimates further. Adjustment for other coronary risk factors had little effect on the results. CONCLUSIONS: At least 80% of major CHD events in middle-aged men can be attributed to the three strongest risk factors. Population-wide control of these factors is crucial for effective CHD prevention. PMID- 14522567 TI - Comparative impact of enalapril, candesartan or metoprolol alone or in combination on ventricular remodelling in patients with congestive heart failure. AB - AIMS: RESOLVD study patients were randomized to candesartan (C), enalapril (E), or C+E. Patients were later randomized to metoprolol CR (M) or placebo. Examine impact of C or E (C/E), C+E, C+M/E+M, C+E+M on ventricular remodelling in heart failure (HF) over 43 weeks. METHODS AND RESULTS: Four hundred and twenty-six of 768 patients receiving C, E, or C+E were randomized to either M or placebo. Patients were New York Heart Association class II-IV, ejection fraction (EF) <0.40 and 6-min walk distance <500 m. Ejection fraction (EF), cardiac volumes, blood pressures, heart rates, and neurohormones were measured. End diastolic volumes changed +29.4+/-6.4 ml for C/E, +16.6+/-10.4 ml for C+E, +19.7+/-6.5 ml for C+M/E+M, and -6.4+/-7.5 ml for C+E+M (P< or =0.01). End systolic volumes changed +22.9+/-5.8 ml for C/E, +11.9+/-9.1 ml for C+E, +6.0+/-5.7 ml for C+E/E+M, and -16.5+/-7.0 ml for C+E+M (P< or =0.001). Ejection fraction changed +0.01+/-0.01 for C/E, +0.01+/-0.01 for C+E, +0.03+/-0.01 for C+M/E+M, and +0.05+/ 0.01 for C+E+M (P< or =0.0001). No significant differences for blood pressure or neurohormones; heart rate for C+M/E+M and C+E+M decreased (P< or =0.01) vs C/E or C+E. CONCLUSION: C+E+M had a modest but beneficial effect on cardiac function compared to the other groups. Combination of C+E+M has potential for providing HF patients with further benefit. PMID- 14522568 TI - N-terminal pro-brain natriuretic peptide. A new gold standard in predicting mortality in patients with advanced heart failure. AB - AIMS: The selection of patients for cardiac transplantation (CTx) is notoriously difficult and traditionally involves clinical assessment and an assimilation of markers of the severity of CHF such as the left ventricular ejection fraction (LVEF), maximum oxygen uptake (peak VO2) and more recently, composite scoring systems e.g. the heart failure survival score (HFSS). Brain natriuretic peptide (BNP) is well established as an independent predictor of prognosis in mild to moderate chronic heart failure (CHF). However, the prognostic ability of NT proBNP in advanced heart failure is unknown and no studies have compared NT proBNP to standard clinical markers used in the selection of patients for transplantation. The purpose of this study was to examine the prognostic ability of NT-proBNP in advanced heart failure and compare it to that of the LVEF, peak VO2 and the HFSS. METHODS AND RESULTS: We prospectively studied 142 consecutive patients with advanced CHF referred for consideration of CTx. Plasma for NT proBNP analysis was sampled and patients followed up for a median of 374 days. The primary endpoint of all-cause mortality was reached in 20 (14.1%) patients and the combined secondary endpoint of all-cause mortality or urgent CTx was reached in 24 (16.9%) patients. An NT-proBNP concentration above the median was the only independent predictor of all cause mortality (chi2=6.03, P=0.01) and the combined endpoint of all cause mortality or urgent CTx (chi2 =12.68, P=0.0004). LVEF, VO2 and HFSS were not independently predictive of mortality or need for urgent cardiac transplantation in this study. CONCLUSION: A single measurement of NT-proBNP in patients with advanced CHF, can help to identify patients at highest risk of death, and is a better prognostic marker than the LVEF, VO2 or HFSS. PMID- 14522569 TI - Lipophilic statins interfere with the inhibitory effects of clopidogrel on platelet function--a flow cytometry study. AB - AIMS: Clopidogrel is a pro-drug which is converted to an active, unstable drug by cytochrome P450 (CYP). The active drug irreversibly blocks one specific platelet adenosine 5'-diphosphate (ADP) receptor (P2Y12). It has been recently suggested that the most abundant human CYP isoform, 3A4, activates clopidogrel. Since certain lipophilic statins (i.e. simvastatin, atorvastatin, lovastatin) are a substrate of CYP3A4, we were interested in potential drug interactions between clopidogrel and statins. METHODS: In patients with coronary artery disease (n=47) in whom clopidogrel treatment was initiated for balloon angioplasty and stent implantation, blood samples were taken at 0, 5 and 48 h after oral administration of clopidogrel (loading dose 300 mg, followed by 75 mg daily). ADP-stimulated (1, 10, 100 micromol/l) expression of P-selectin (CD62P) on platelets was measured by flow cytometry, and used as a marker for the antiplatelet effect of clopidogrel. RESULTS: Pre-treatment with statins (atorvastatin, simvastatin) reduced significantly (10 micromol/l ADP stimulation) the inhibitory effects of clopidogrel during the loading phase (relative reduction after 5 h 29.3%) and, to a lesser extent during the maintenance phase (relative reduction after 48 h 16.6%). In addition we found a considerable individual heterogeneity in the response and three patients (6%) were identified in whom clopidogrel exerted almost no effect. CONCLUSION: Certain statins which are substrates of the CYP3A4 isoform competitively inhibit the metabolic activation of clopidogrel. As a result the relative clopidogrel induced platelet inhibition (P-selectin expression) is diminished--but still there is a relative clopidogrel effect of more than 80% in the maintenance phase. It may be reasonable to test the therapeutic efficacy of clopidogrel in those patients who require long-term treatment. PMID- 14522570 TI - A short delay from out of hospital cardiac arrest to call for ambulance increases survival. AB - AIM: To describe the relative impact on survival of the delay from estimated time of collapse to call for an ambulance among patients who suffer from a bystander witnessed out of hospital cardiac arrest of a cardiac aetiology. METHODS: A majority of all ambulance organizations in Sweden (covering 85% of Sweden inhabitants) participate in a National survey of out of hospital cardiac arrest. RESULTS: In all there were 9340 patients with a bystander witnessed cardiac arrest of a cardiac aetiology in whom cardiopulmonary resuscitation (CPR) was attempted participating in this survey. Survival at one month among patients with a delay between estimated time of collapse and call for ambulance of < or =4 min (median) was 6.9% versus 2.8% among patients with a median of >4 min (P<0.0001). When adjusting for age, sex, initial rhythm, estimated interval between collapse and start of CPR, place of arrest and the interval between call for ambulance and arrival of the rescue team, the odds ratio for survival was 0.70 (0.95% CI. 0.58 0.84) per unit increase of the natural logarithm of delay in minutes between collapse and call. CONCLUSION: Among patients with a bystander witnessed out of hospital cardiac arrest of a cardiac aetiology increased delay from estimated time of collapse to call for an ambulance decreased the chance of survival. PMID- 14522571 TI - Patients with familial hypercholesterolaemia show enhanced spontaneous chemokine release from peripheral blood mononuclear cells ex vivo. Dependency of xanthomas/xanthelasms, smoking and gender. AB - AIMS: Familial hypercholesterolaemia (FH) is associated with increased risk of premature atherosclerosis and coronary artery disease (CAD). However, onset of clinically manifested CAD varies widely among patients with heterozygous FH, and we hypothesized that inflammatory mediators such as chemokines could contribute to atherogenesis in these patients. METHODS AND RESULTS: We compared peripheral blood mononuclear cells (PBMCs) from FH patients with an identical mutation with PBMCs from sex- and age-matched healthy controls with respect to spontaneous and oxidized low density lipoprotein (oxLDL)-stimulated release of chemokines. Our main findings were: (1) PBMCs from FH patients spontaneously released significantly higher levels of macrophage inflammatory protein (MIP)-1alpha, MIP 1beta and interleukin (IL)-8, and had a significantly lower oxLDL-stimulatory ratio for MIP-1alpha and MIP-1beta than cells from healthy controls. (2) Spontaneous release of these chemokines correlated positively and stimulatory ratio correlated negatively with plasma concentrations of total and LDL cholesterol. (3) Among FH patients, release of MIP-1alpha, MIP-1beta and IL-8 from PBMCs varied with the presence of xanthomas/xanthelasms, smoking and gender. (4) In vitro studies showed that FH serum but not control serum was able to induce enhanced spontaneous release of chemokines in PBMCs from both FH patients and control subjects. CONCLUSIONS: Our data may suggest that a pathophysiological consequence of FH is enhanced chemokine responses, which in turn may promote recruitment and activation of leukocytes within the vessel wall, contributing to atherosclerosis as well as to the different phenotypes in these patients with an identical FH mutation. PMID- 14522572 TI - Intra-aortic balloon counterpulsation in US and non-US centres: results of the Benchmark Registry. AB - AIMS: To examine differences in patient characteristics and outcomes in 19636 patients enrolled in the USA and 3027 patients enrolled in other countries undergoing intra-aortic balloon pump (IABP) counterpulsation. METHODS AND RESULTS: Indications for IABP use; a larger percentage of US patients were identified as 'early support and stabilization for angiography or angioplasty' (21.1% US vs 11.8% non-US), and 'pre-operative support for high-risk CABG' (15.9% vs 6.6%). A smaller percentage of US patients vs non-US patients were identified as 'weaning from cardiopulmonary bypass' (14.3% vs 28.2%), and 'refractory ventricular failure' (6.2% vs 9.8%). One out of five patients in both groups was listed as 'cardiogenic shock' (18.9% US vs 20.2% non-US). All cause, risk adjusted, in-hospital mortality (20.1% vs 28.7%; P<0.001), and mortality with IABP in place (10.8% vs 18.0%; P<0.001) were lower at US vs non-US sites. In both US and non-US institutions, IABP associated complication rates, such as IABP related mortality (0.05% vs 0.07%), major limb ischaemia (0.9% vs 0.8%), and severe bleeding (0.9% vs 0.8%), were low. CONCLUSIONS: IABP counterpulsation is deployed at an earlier clinical stage in US patients. Mortality rates are higher for non-US patients, particularly for patients with non-surgery cardiac interventions, even after adjusting for risk factors. Complication rates were low. Physicians should therefore not be reluctant to use IABP in high-risk patients undergoing cardiac procedures. PMID- 14522573 TI - Heart rate variability in patients with Brugada syndrome in Thailand. AB - AIMS: Since patients with Brugada syndrome usually have symptoms at nighttime, we hypothesize that changes in autonomic modulation have an important role in the occurrence of the ventricular fibrillation episodes. The objective of this study was to determine the changes in heart rate variability (HRV) in patients with Brugada syndrome compared to asymptomatic subjects with Brugada ECG and controls. METHODS AND RESULTS: We studied 17 patients with Brugada syndrome, 10 asymptomatic subjects with Brugada ECG and 45 controls. Patients with Brugada syndrome and asymptomatic subjects with Brugada ECG underwent echocardiography, exercise stress testing, 24-h Holter monitoring, signal-averaged ECG. Patients with Brugada syndrome also underwent coronary angiography and electrophysiologic study. Time domain and frequency domain HRV analysis were performed at daytime and nighttime. The results of this study showed that patients with Brugada syndrome had lower HRV or lower vagal tone at night compared to the controls. They also had lower heart rate during the day and higher during the night compared to asymptomatic subjects and the controls. CONCLUSION: Patients with Brugada syndrome had low heart rate variability at night which may predispose to the occurrence of VF episodes. PMID- 14522574 TI - Suspected pacemaker or defibrillator transvenous lead infection. Prospective assessment of a TEE-guided therapeutic strategy. AB - AIMS: The aim of this prospective study was to assess the clinical value of a management strategy principally based on the results of multiplane transoesophageal echocardiography (TEE) in patients with suspected lead infection. METHODS AND RESULTS: Seventy-seven consecutive patients were included. Based on Duke's modified criteria, PTLI was considered as definite in 54 patients (70%) and possible in 23 patients (30%). Nineteen patients with a diagnosis of possible infection, as defined by bacteraemia without abnormal TEE images and without evidence of pacemaker pocket infection, were treated by antibiotics alone. In all other cases, the pacing material was totally removed. During a mean follow-up time of 3.1+/-2.5 years, 21 patients (27%) died, mostly from cardiovascular causes. Only one patient died from infection and there was only one case of delayed infection recurrence in an other localization. No significant differences in outcome were observed between explanted and non-explanted patients. CONCLUSIONS: The results observed confirm that early and total explantation of pacing material has to be done in patients with bacteraemia and abnormal images at TEE. But conversely conservation of the pacing system can be proposed to patients with bacteraemia but without abnormal images at TEE provided prolonged antibiotic treatment is given. PMID- 14522576 TI - Population-based cardiovascular risk prediction. PMID- 14522575 TI - The effect of reduced blood-flow on the coronary wall temperature. Are significant lesions suitable for intravascular thermography? AB - AIMS: The purpose of this study was to investigate the relation between acute coronary flow reduction and arterial wall temperature. METHODS AND RESULTS: Five pigs with normal coronary arteries were catheterized. Arterial wall temperature was studied with a thermographic system that uses a 4-thermistor sensor tip. Flow velocity was studied at the same time and place with the temperature measurements, using a Doppler wire. In order to modify the coronary flow, a balloon was gradually inflated proximally to the thermographic sensors. Temperature differences and flow velocities were simultaneously recorded. Flow velocities above an average peak velocity (APV) of 9 cm/s were associated with unaffected temperature measurements. At flow velocities around 4 cm/s, the wall temperature was increased (deltaT=0.015+/-0.005 degrees C, P approximately 0.05), following the heart-rate. When flow velocity dropped further below this value, the local wall temperature was logarithmically increased to a maximum value observed at total vessel occlusion (deltaT=0.188+/-0.023 degrees C, P<0.001). CONCLUSION: The reduction of coronary flow has an effect on the arterial wall temperature. This effect however, appears only below a critical threshold of APV and in a logarithmic fashion. Above this threshold, temperature measurements should be unaffected from flow reductions and related to the regional temperature heterogeneity. PMID- 14522578 TI - Closure of atrial septal defect: is the debate over? PMID- 14522580 TI - Long distance transport for primary angioplasty vs immediate thrombolysis in acute myocardial infarction (PRAGUE-2 trial). PMID- 14522581 TI - Rhein induces apoptosis in HL-60 cells via reactive oxygen species-independent mitochondrial death pathway. AB - Rhein is an anthraquinone compound enriched in the rhizome of rhubarb, a traditional Chinese medicine herb showing anti-tumor promotion function. In this study, we first reported that rhein could induce apoptosis in human promyelocytic leukemia cells (HL-60), characterized by caspase activation, poly(ADP)ribose polymerase (PARP) cleavage, and DNA fragmentation. The efficacious induction of apoptosis was observed at 100 microM for 6h. Mechanistic analysis demonstrated that rhein induced the loss of mitochondrial membrane potential (DeltaPsi(m)), cytochrome c release from mitochondrion to cytosol, and cleavage of Bid protein. Rhein also induced generation of reactive oxygen species (ROS) and the phosphorylation of c-Jun N-terminal kinase (JNK) and p38 kinase. However, these actions seem not to be associated with the apoptosis induction because antioxidants including N-acetyl cysteine (NAC), Tiron, and catalase did not block rhein-induced apoptosis, although they could block the generation of ROS and the phosphorylation of JNK and p38 kinase. Our data demonstrate that rhein induces apoptosis in HL-60 cells via a ROS-independent mitochondrial death pathway. PMID- 14522582 TI - Effects of zinc on factor I cofactor activity of C4b-binding protein and factor H. AB - Complement inhibition is to a large extent achieved by proteolytic degradation of activated complement factors C3b and C4b by factor I (FI). This reaction requires a cofactor protein that binds C3b/C4b. We found that the cofactor activity of C4b binding protein towards C4b/C3b and factor H towards C3b increase at micromolar concentrations of Zn(2+) and are abolished at 2 mM Zn(2+) and above. 65Zn(2+) bound to C3b and C4b molecules but not the cofactors or FI when they were immobilized in a native form on a nitrocellulose membrane. Zn(2+) binding constants for C3met (0.2 microM) and C4met (0.1 microM) were determined using fluorescent chelator. It appears that higher cofactor activity at low zinc concentrations is due to an increase of affinity between C4b/C3b and cofactor proteins as assessed by surface plasmon resonance. Inhibition of the reaction seen at higher concentrations is due to aggregation of C4b/C3b. PMID- 14522583 TI - The requirement of ammonium or other cations linked with p-cresol sulfate for cross-reactivity with a peptide of myelin basic protein. AB - Urinary myelin basic protein-like material (MBPLM), so designated because of its immunoreactivity with a polyclonal antibody directed against a cryptic epitope located in residues 83-89 of myelin basic protein (MBP), exists in humans normally but increases in concentration in patients with multiple sclerosis who have progressive disease. Given its possible role in reflecting events of neural tissue destruction occurring in multiple sclerosis, urinary MBPLM is a candidate surrogate marker for this phase of the disease. Previously, it has been demonstrated that p-cresol sulfate (PCS) is the dominant component of MBPLM; however, another component(s) was essential in enabling p-cresol sulfate to have molecular mimicry with MBP peptide 83-89 detected by immunoreactivity. In the present investigation, this remaining component(s) was characterized by a combination of high performance size exclusion chromatography followed by nuclear magnetic resonance spectroscopy and shown to be ammonium. The monovalent cation ammonium could be substituted in vitro by several different monovalent and divalent cations, most notably zinc, in restoring to deprotonated p-cresol sulfate its immunoreactivity as MBPLM. These findings indicate the basis for the unexpected molecular mimicry between an epitope of an encephalitogenic protein and a complex containing a small organic molecule, p-cresol sulfate. Furthermore, the reaction of either ammonium or other cations with p-cresol sulfate may represent an in vivo process directly related to damage of axonal membranes. PMID- 14522584 TI - Abnormal growth of polyamine-deficient Escherichia coli mutant is partially caused by oxidative stress-induced damage. AB - Polyamines participate in numerous cellular processes and are required for normal cell growth in Escherichia coli. In this study, we constructed a new polyamine deficient E. coli mutant and investigated the physiological function of polyamines during normal aerobic growth conditions. We showed that the requirement for sulfur-containing, branched chain, and aromatic amino acids, which was exhibited in the sodA sodB double mutant faced with severe oxidative stress, was also true of the polyamine-deficient mutant during normal aerobic cell growth. Sorbitol, sucrose, mannose, 1,2-dihydroxybenzene-3,5-disulfonic acid (Tiron), an antioxidant that functions as an oxygen radical scavenger including z.rad;O(2)(-), and thiamine partially relieved the cell growth defect caused by polyamine depletion in a dose-dependent manner. As was the case for the cells treated with paraquat, the mutant had an elongated shape compared with the polyamine-proficient wild type. Decreased aeration also relieved the cell growth defect of the polyamine-deficient mutant. Finally, we confirmed that chloromethyl 2('),7(')-dichlorofluorescin diacetate (DCFH-DA), which is oxidized in a fluorescent product in the presence of various oxidants, also fluoresce in the polyamine-deficient cells. These results showed that abnormal growth of the polyamine-deficient E. coli mutant results partially from oxidative stress induced damage and the mutant thus exhibits the requirement for antioxidant or specific nutritional amino acid during normal aerobic growth. PMID- 14522585 TI - Product formation and kinetic simulations in the pH range 1-14 account for a free radical mechanism of peroxynitrite decomposition. AB - The yields of nitrate and nitrite from decomposition of peroxynitrite in phosphate buffer at 37 degrees C were determined in the pH range 1-14. The NO(2)( )/NO(3)(-) yields showed a stepwise variation with pH, with inflection points at approximately pH 3.1, 5.8, 6.8, 8.0, and 11.9. Nitrite formation increased strongly above pH 7 at the expense of nitrate, but above pH 12 nitrate again became the major product (80% at pH 14). At this pH, the Arrhenius parameters were E(a)=24.1+/-0.2kcal mol(-1) and A=(4.9+/-1.3)x10(12)s(-1). The yields of NO(2)(-), NO(3)(-), and O(2) measured at pH 5.8, 7.4, and 8.5 as a function of the initial peroxynitrite concentration (50-1000 microM) were linear only at pH 5.8. In the presence of carbon dioxide, oxygen production at pH 7.5 and pH 10 was found to be linear on the CO(2) concentration. The experimental observations were satisfactorily reproduced by kinetic simulations including principal component analyses. These data strongly suggest that the chemistry of peroxynitrite is exclusively mediated by z.rad;NO(2) and HO(z.rad;) radicals in the absence, and by z.rad;NO(2) and CO(3)(z.rad;-) radicals in the presence of CO(2). PMID- 14522586 TI - 5alpha-reduced C21 steroids are substrates for human cytochrome P450c17. AB - The 5alpha-reduction of testosterone in target tissues is a key step in androgen physiology; however, 5alpha-reduced C(19) steroids are sometimes synthesized in testis via a pathway that does not involve testosterone as an intermediate. We studied the metabolism of 5alpha-reduced C(21) steroids by human cytochrome P450c17 (hCYP17), the enzyme responsible for conversion of C(21) steroids to C(19) steroids via its 17alpha-hydroxylase and 17,20-lyase activities. hCYP17 17alpha-hydroxylates 5alpha-pregnan-3,20-dione, but little androstanedione is formed by 17,20-lyase activity. hCYP17 also 17alpha-hydroxylates 5alpha-pregnan 3alpha-ol-20-one and the 5alpha-pregnan-3alpha,17alpha-diol-20-one intermediate is rapidly converted to androsterone by 17,20-lyase activity. Furthermore, 5alpha pregnan-3alpha,17alpha-diol-20-one is a better substrate for the 17,20-lyase reaction than the preferred substrate 17alpha-hydroxypregnenolone and cytochrome b(5) stimulates androsterone formation only 3-fold. Both 5alpha-pregnan-3alpha-ol 20-one and 5alpha-pregnan-3alpha,17alpha-diol-20-one bind to hCYP17 with higher affinity than does progesterone. We conclude that 5alpha-reduced, 3alpha-hydroxy C(21) steroids are excellent, high-affinity substrates for hCYP17. The brisk metabolism of 5alpha-pregnan-3alpha,17alpha-diol-20-one to androsterone by CYP17 explains how, when 5alpha-reductases are present, the testis can produce C(19) steroids androsterone and androstanediol from 17alpha-hydroxyprogesterone without the intermediacy of androstenedione and testosterone. PMID- 14522587 TI - Methotrexate is a novel inducer of rat liver and intestinal sulfotransferases. AB - The antifolate drug methotrexate (MTX) is commonly used in the treatment of different types of cancers and autoimmune diseases. In the present investigation, MTX is shown to be a novel xenobiotic inducer of rat liver and intestinal sulfotransferases (STs). STs are phase II drug metabolizing enzymes. ST induction by hormones and other endogenous molecules has been relatively well studied. Xenobiotic drug induction of STs is not well known. Our enzyme assay, Western blot, and reverse transcription polymerase chain reaction (RT-PCR) results demonstrated that protein and mRNA expressions of aryl sulfotransferase (AST-IV) and hydroxysteroid sulfotransferase (STa) were induced in liver and intestine of male and female rats following MTX treatment at three different doses (0.04, 0.2, and 1mg/kg/day) for 2 weeks. Intestinal inductions were found to be much greater than those found in liver. MTX is the first antifolate and apoptosis-inducing drug to show induction of STs. PMID- 14522588 TI - Challenges and complexities of alpha-synuclein toxicity: new postulates in unfolding the mystery associated with Parkinson's disease. AB - The discovery of two missense mutations in alpha-synuclein gene and the identification of the alpha-synuclein as the major component of Lewy bodies and Lewy neurites have imparted a new direction in understanding Parkinson's disease. Now that alpha-synuclein has been implicated in several neurodegenerative disorders makes it increasingly clear that aggregation of alpha-synuclein is a hallmark feature in neurodegeneration. Although little has been learned about its normal function, alpha-synuclein appears to be associated with membrane phospholipids and may therefore participate in a number of cell signaling pathways. Here, we review the localization, structure, and function of alpha synuclein and provide a new hypothesis on, (a) the disruption in the membrane binding ability of synuclein which may be the major culprit leading to the alpha synuclein aggregation and (b) the complexity associated with nuclear localization of alpha-synuclein and its possible binding property to DNA. Further, we postulated the three possible mechanisms of synuclein induced neuronal degeneration in Parkinson's disease. PMID- 14522589 TI - A hyperthermostable novel protein-disulfide oxidoreductase is reduced by thioredoxin reductase from hyperthermophilic archaeon Pyrococcus horikoshii. AB - A redox protein gene (PH0178) with high sequence homology to a glutaredoxin from Pyrococcus furiosus and a thioredoxin reductase homologue gene (PH1426) were found in the genome sequence of Pyrococcus horikoshii. These two genes were cloned and the corresponding expressed proteins were characterized. The redox protein from PH0178 had strong thioredoxin-like activity, but no glutaredoxin activity. The protein from PH1426 had some reductase activity against thioredoxin from Escherichia coli as well as the redox protein (PH0178). The protein from PH1426 was a typical, homodimeric flavoprotein. These results indicate that the redox protein (PH0178) is not a glutaredoxin but, rather, a new protein-disulfide oxidoreductase that is involved in a thioredoxin-like system with thioredoxin reductase (PH1426) in P. horikoshii. The redox protein and thioredoxin reductase retained their full activities for over 1h at 100 degrees C. The redox potential of the redox protein was similar to that of thioredoxin from E. coli and lower than that of glutathione. Site-directed mutagenesis studies revealed that the active site of the redox protein corresponds to a CPYC sequence, located in the middle of the sequence. PMID- 14522590 TI - Peroxisome proliferator-activated receptor gamma ligands attenuate immunological symptoms of experimental allergic asthma. AB - Asthma is characterized by a predominant T(H)2 type immune response to airborne allergens. Controlling T(H)2 cell function has been proposed as therapy for this disease. We show here that ligands for the nuclear receptor peroxisome proliferator activated receptor (PPAR)gamma significantly reduced the immunological symptoms of allergic asthma in a murine model of this disease. A PPARgamma ligand, 15-deoxy-delta(12,14)-prostaglandin J(2), significantly inhibited production of the T(H)2 type cytokine IL-5 from T cells activated in vitro. More importantly, in a murine model of allergic asthma, mice treated orally with ciglitazone, a potent synthetic PPARgamma ligand, had significantly reduced lung inflammation and mucous production following induction of allergic asthma. T cells from these ciglitazone treated mice also produced less IFNgamma, IL-4, and IL-2 upon rechallenge in vitro with the model allergen. Our results suggest that ligands for PPARgamma may be effective treatments for asthmatic patients. PMID- 14522591 TI - Exploiting the conformational flexibility of leghemoglobin: a framework for examination of heme protein axial ligation. AB - We have exploited the intrinsic conformational flexibility of leghemoglobin to reengineer the heme active site architecture of the molecule by replacement of the mobile His61 residue with tyrosine (H61Y variant). The electronic absorption spectrum of the ferric derivative of H61Y is similar to that observed for the phenolate derivative of the recombinant wild-type protein (rLb), consistent with coordination of Tyr61 to (high-spin) iron. EXAFS data clearly indicate a 6 coordinate heme geometry and a Fe-O bond length of 185pm. MCD and EPR spectroscopies are consistent with this assignment and support ligation by an anionic (tyrosinate) group. The alteration in heme ligation leads to a 148mV decrease in the reduction potential for H61Y (-127+/-5mV) compared to rLb and destabilisation of the functional oxy-derivative. The results are discussed in terms of our wider understanding of other heme proteins with His-Tyr ligation. PMID- 14522592 TI - The Escherichia coli ynfEFGHI operon encodes polypeptides which are paralogues of dimethyl sulfoxide reductase (DmsABC). AB - The ynfEFGHI operon is a paralogue of the Escherichia coli dmsABC operon. ynfE and ynfF are paralogues of dmsA. ynfG and ynfH are paralogues of dmsB and dmsC, respectively. YnfI (dmsD) has no dms paralogue. YnfE/F and YnfG could be detected by immunoblotting with anti-DmsAB antibodies when expressed under the control of a tac or dms promoter. Cells harbouring ynfFGH on a multicopy plasmid supported anaerobic growth with dimethyl sulfoxide (DMSO) as respiratory oxidant in a dmsABC deletion, suggesting that YnfFGH forms a heterotimeric enzyme complex similar to DmsABC. Exchange of DmsC by YnfH (DmsAB-YnfH) resulted in membrane localization, anaerobic growth on DMSO, and binding of 2-n-heptyl 4 hydroxyquinoline-N-oxide, indicating that YnfH was a competent anchor. YnfG can also replace DmsB as the electron transfer subunit and assembled [Fe-S] clusters as judged by electron paramagnetic resonance spectroscopy. YnfE and/or YnfF could not form a functional complex with DmsBC and expression of YnfE prevented the accumulation of YnfFGH. PMID- 14522593 TI - Enzymatic and transcriptional regulation of human ecto-ATPase/E-NTPDase 2. AB - We have characterized the regulation of expressed human ecto-ATPase (E-NTPDase 2), a cell surface integral membrane glycoprotein. Ecto-ATPase activity is inhibited by parameters that decrease membrane protein interaction, i.e., detergents and high temperatures. These inhibitory effects are overcome when membranes are pretreated with concanavalin A or chemical cross-linking agents that increase the amounts of ecto-ATPase oligomers. Cross-linking agents also abrogate substrate inactivation of the ecto-ATPase, a unique characteristic of the enzyme. These effects indicate that the magnitude of negative substrate regulation is dependent on quaternary structures of the protein, which likely involves interaction of transmembrane domains. The importance of transmembrane domains of ecto-ATPase in activity modulation is demonstrated further by the stimulatory effect of digitonin, a steroid glycoside that preferentially interacts with cholesterol in the membranes but does not promote oligomer formation. These results indicate that ecto-ATPase activity is regulated by a multitude of mechanisms, some of which may have physiological significance. Ecto ATPase is also susceptible to transcriptional regulation. Ecto-ATPase gene expression is increased in a human hepatoma whereas it is undetectable in the normal liver. PMID- 14522594 TI - Ion transport proteins and aquaporin water channels in the kidney of amphibians from different habitats. AB - Amphibians are known to spend part of their life on land and return to water to reproduce. However, some urodeles spend their entire life in water, while others succeed in completely avoiding water even during reproduction. Osmoregulatory mechanisms must therefore be different in the diverse environmental conditions of their respective life histories. The architecture of the kidney is similar in all amphibians; as a consequence the ion-water equilibrium must be regulated in the different environmental conditions. We investigated the immunolocalisation of Na(+)/K(+)/Cl(-) cotransport proteins, sodium pump and water-channel proteins (aquaporins) in aquatic Amphiuma means means, Rana dalmatina, a species that returns to water to reproduce, and Speleomantes genei, a completely terrestrial species. The investigation was carried out with immunohistochemical methods using antibodies to Na(+)/K(+)/Cl(-) cotransport protein NKCC1 T4, Na(+)/K(+)ATPase alpha-subunit, water-channel aquaporin 3 and the inner mitochondrial membrane (AMA). Cotransport proteins and sodium pump, involved in ion reabsorption, are widely distributed in A. means and R. dalmatina and confined to the distal segment in S. genei; conversely water channels, involved in water reabsorption, are limited to the collecting duct in A. means and R. dalmatina and distributed in the proximal and collecting ducts in S. genei. PMID- 14522595 TI - Ginseng increases intestinal elimination of albendazole sulfoxide in the rat. AB - Herbal products show potential drug interactions, some of them with adverse effects. The main aim of this work was to study the effect of Panax ginseng on the intestinal elimination of the benzimidazole derivative albendazole sulfoxide (ABZSO). An upper small intestine segment was isolated and perfused in situ with saline, while ABZSO solution (10 mg/kg i.v.) was administered intravenously. Blood samples and intestinal secretion were collected over 60 min and analysed by HPLC. The intestinal clearance of ABZSO was 0.106+/-0.010 ml/min. Systemic co administration of ginseng (10 mg/kg i.v.) increased significantly (P<0.05) the clearance of ABZSO (0.132+/-0.005 ml/min). The increase in ABZSO elimination could be the result of the effect of ginseng on metabolic pathways. These results highlight the interactions between herbal products (sometimes dietary constituents) and drugs such as benzimidazoles, since ginseng modifies the luminal clearance of this anthelminthic drug and could potentially interfere with drugs that undergo the same intestinal processes. PMID- 14522596 TI - Embryonic exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin in chickens: effects of dose and embryonic stage on hatchability and growth. AB - Chicken embryos (Gallus domesticus) were injected with 0, 8, 20 or 50 ng tetrachlorodibenzo-p-dioxin (TCDD) per egg at embryonic day (ED) 4, 8 or 12 to investigate the effects of differential periods of sensitivity to TCDD exposure. At hatch, all chicks were weighed, sexed and examined macroscopically to identify possible malformations. Liver, bursa, heart and spleen masses were recorded from a number of chicks. The remaining chicks were raised until 6 weeks of age and body and organ masses, plasma concentrations of thyroid hormones, triglycerides and glucose were measured. Dose and stage during embryonic development at which injection was performed affected hatchability. Fifty nanogram of TCDD was highly toxic for 4-day-old chicken embryos. TCDD was less toxic for chicken embryos of 8 and especially 12-days old. One-day-old chick and organ weights were not different between TCDD doses at all injection days. However, injection performed at ED4 or ED8 with 20 and 50 ng, respectively, significantly depressed post-hatch body mass gain. Moreover, body mass gain in males was more depressed than in females. The delayed growth in TCDD treated chickens was accompanied by changes in T(3)/T(4) ratio that at some ages were significantly higher compared to control animals. No pronounced changes in plasma triglycerides or glucose concentrations during postnatal life were observed. Absolute and relative organ masses of 6-week-old chickens showed no remarkable changes. PMID- 14522597 TI - Oxidative stress in liver and brain of the hatchling chicken (Gallus domesticus) following in ovo injection with TCDD. AB - 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) was injected into chicken eggs prior to incubation to study possible mechanisms of toxicity and teratogenicity. One of the suggested mechanisms of teratogenicity is oxidative stress. Eggs were injected simultaneously with TCDD and cotreatment compounds in an attempt to prevent oxidative stress or to block cytochrome P450 activity. Indicators of oxidative stress were assessed in livers and brains of hatchling chicks. In ovo, exposure to TCDD caused significant effects on indicators of oxidative stress in liver, but not in the brain of the hatchling chicks. TCDD did not significantly affect superoxide production. In liver, TCDD treatment caused a decrease in glutathione content and glutathione peroxidase activity and an increase in the ratio of oxidized to reduced glutathione. TCDD increased the susceptibility to lipid peroxidation and oxidative DNA damage in liver. Administration of the antioxidants vitamin E and vitamin A provided partial protection against TCDD induced oxidative stress in liver. The lack of effect of TCDD in chicken brain could be due to the low cytochrome P4501A activity in this tissue and little accumulation of TCDD in brain compared to liver. Phenytoin, a known inducer of oxidative stress, caused a decrease in glutathione content and an increase in susceptibility to lipid peroxidation in both liver and brain and increased oxidative DNA damage in brain. Responsiveness varied among individual animals, but measures of the oxidative stress were correlated. PMID- 14522599 TI - Purification and characterization of an antibacterial protein in the skin secretion of rockfish Sebastes schlegeli. AB - An antibacterial protein in the skin secretion of rockfish (Sebastes schlegeli) was purified by lectin affinity chromatography on Con A-Sepharose and gel filtration on TSKgel G3000SW. The antibacterial protein featured the high molecular mass and selective action against Gram-negative bacteria. The molecular mass of the protein was estimated to be approximately 150 kDa in gel filtration and approximately 75 kDa by SDS-PAGE, suggesting that it is dimeric. The antibacterial principle was an acidic glycoprotein with pI 4.5, 3.4% reducing sugar and 2.8% amino sugar. Its sugar chains had N-type (high mannose-type) oligosaccharide and sialic acid components. It inhibited strongly the growth of Aeromonas salmonicida, Photobacterium damselae and Shewanella putrefaciens with a minimum inhibitory concentration (MIC) of approximately 3 microg/ml, and moderately the growth of Vibrio parahaemolyticus and A. hydrophila with a MIC of 12.5 microg/ml and 25 microg/ml, respectively. The values of the minimum bactericidal concentration were almost equivalent to those of MIC. The potent sensitivity against virulent pathogens such as A. hydrophila, A. salmonicida and P. damselae may contribute considerably to the innate host defense mechanism to combat microbes on the mucosal surfaces of the rockfish. PMID- 14522598 TI - Mechanisms of TCDD-induced abnormalities and embryo lethality in white leghorn chickens. AB - The toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related compounds in birds has been well-established in laboratory and field studies. Observed effects of TCDD and related chemicals in birds include developmental deformities, reproductive failure, liver damage, wasting syndrome and death. The mechanism of action of TCDD at the cellular level is primarily mediated through the aryl hydrocarbon receptor (AhR). However, the mechanism of toxic action at the organism level is poorly understood. In this study, the role of radical oxygen species and mixed function oxidize (MFO; cytochrome P4501A) in the mechanism of TCDD-induced abnormalities and lethality were examined by co-injecting radical scavengers and an MFO inhibitor (piperonyl butoxide). Egg injection studies were conducted to determine if in ovo TCDD exposure can cause oxidative stress in white leghorn chicken eggs. Test agents were injected into the yolk prior to incubation. Treatments included TCDD (150 ng/kg), triolein (vehicle control), and various co-treatments including MnTBAP (a mimetic of superoxide dismutase), piperonyl butoxide, piroxicam, vitamin A acetate, and vitamin E succinate. Phenytoin, which is known to cause teratogenesis through oxidative stress was used as a positive control. Eggs were incubated until hatch and then the following parameters were assessed: mortality, hatching success, abnormalities, weights for whole body, liver, heart and brain, and biochemical endpoints for oxidative stress. As a measure of exposure, concentrations of TCDD and ethoxyresorufin-O-deethylase (EROD) activities were measured in tissues of hatchlings. While greater mortality and abnormalities were observed in the TCDD treatment groups, the number of the replicates were not great enough to detect statistically significant differences in abnormality rates for the co-treatments. Some of the observed developmental abnormalities included edema, liver necrosis and bill, eye and limb deformities with TCDD treatments, bill and brain deformities with phenytoin treatments, eye abnormalities with Vitamin E treatments, and abnormal feather pigmentation with piperonyl butoxide treatments. PMID- 14522600 TI - Fatty acid metabolism in neonatal chickens (Gallus domesticus) treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) or 3,3',4,4',5-pentachlorobiphenyl (PCB-126) in ovo. AB - Treatment of chickens as pre-incubation embryos with TCDD or PCB-126 altered fatty acid concentrations in their plasma 21 days later, compared with their oil vehicles (sunflower and corn oils, respectively). TCDD increased the concentrations of total fatty acids, lipid classes (phospholipids and cholesterol ester), fatty acid families (saturated, n-7 and n-6), and many specific fatty acids. The only fatty acid concentrations decreased by TCDD treatment were those of cholesterol ester fatty acids 20:3n3 and 24:6n3 and overall plasma 24:6n3. In contrast, PCB-126 treatment decreased total phospholipid, saturated and plasmogen fatty acid concentrations with generally decreasing trends in specific fatty acid concentrations. However, both TCDD and PCB-126 treatments increased total 22:1n9 and decreased 24:6n3 concentrations compared with their respective vehicles. The potential relationship between those fatty acid concentrations altered by toxicant treatment and alterations in brain symmetry was then examined using correlation analysis. Several fatty acid concentrations were significantly correlated with differences in brain morphology between the right and left hemispheres and these potential associations were different between toxicant and vehicle. PMID- 14522601 TI - Sex differences in the pharmacokinetics of pioglitazone in rats. AB - Clinical studies have suggested that pioglitazone, an insulin sensitizer, has a stronger effect in women than in men. To determine the sex difference in the pharmacokinetics of pioglitazone, we examined the plasma and white adipose tissue levels of pioglitazone and its active metabolites (M-II, M-III and M-IV) in male and female rats treated with a single or repeated oral administration of pioglitazone (10 mg/kg). The AUCs of pioglitazone (149.6+/-22.6 vs. 103.3+/-14.0 microg.h/ml; P<0.01), M-III (31.4+/-8.1 vs. 20.2+/-4.7 microg.h/ml; P<0.05) and M IV (41.9+/-15.5 vs. 14.1+/-1.6 microg.h/ml; P<0.01) were larger in female rats than in male rats, but the levels of M-II were similar. Any of the compounds did not accumulate in plasma after repeated administration. According to kinetic model analysis, the apparent elimination rate of pioglitazone and the formation rate of M-II were faster in male rats than in female rats. No significant sex difference was found in the tissue-to-plasma concentration ratios of pioglitazone or its active metabolites in white adipose tissue. These results suggest that there are sex differences in the plasma levels of pioglitazone and some of its active metabolites and that those differences are reflected in differences in white adipose tissue levels. PMID- 14522602 TI - Fetal and maternal tissue distribution of the new fluoroquinolone DW-116 in pregnant rats. AB - We investigated maternal and fetal tissue distribution of DW-116, a newly developed fluoroquinolone with a broad antibacterial spectrum against both G(+) and G(-) bacteria, in pregnant rats. After oral administration of [14C]-DW-116 (labeled 1 mg and unlabeled 500 mg/kg) to female rats on the 18th day of gestational, groups of three rats were killed at various time points up to 24 h, and plasma and tissues were collected, processed and analyzed. [14C]-DW-116 was rapidly absorbed, and distributed into the maternal and fetal tissues, and it declined in a biphasic manner with elimination half-lives (t(1/2)) of 10-15 h and mean residence times (MRT(0-24 h)) of 4-9 h. The radioactivity in most tissues of both dams and fetus reached its peak within 1 h and radioactivity levels of up to 10-25% of the peak level were maintained until 24 h after dosing. Among various tissues, the radioactivity in the maternal lungs was the highest (27 times that of plasma) at the C(max). Radioactivity in other tissues including liver, kidney, heart, lung, brain, spleen, mammary gland, placenta, ovary and uterus was higher than that in the maternal plasma (one- to three-fold). The tissue-to-plasma partition coefficient (K(p), AUC(0-24 h,tissue)/AUC(0-24 h,plasma)) of [14C]-DW 116 in maternal tissues was highest in the lung (K(p)=3.7), followed by the spleen (2.2), kidney (2.0), liver (1.8), heart (1.5), placenta (1.3), brain (1.3), ovary (1.1), uterus (1.1), and mammary gland (1.0). The tissue-to-plasma partition coefficient values in fetal tissues were heart (K(p)=2.2), kidney (2.1), liver (1.9), lung (1.6) and brain (1.4). When lactating rats were given a single oral dose of [14C]-DW-116, the radioactivity was rapidly secreted into the milk with K(p) of 1.7 at T(max) (0.5 h). These results indicate that DW-116 or its related metabolite(s) rapidly cross the blood-placenta and blood-milk barrier, extensively distribute into the fetal tissues, and are eliminated from the body in a prolonged manner. This study sheds insights into the maternal and fetal tissue distribution of DW-116 and will be useful for assessing both therapeutic and toxicological relevance of DW-116 in pregnant subjects. PMID- 14522603 TI - Conversion disorder and its subtypes: a need for a reclassification. AB - Four subtypes of conversion disorder were described in DSM-IV. There are few publications concerning studies aimed at separating the subtypes of the conversion disorder. Usually, pseudoseizures are in focus and attempts are made to differentiate these seizures from other disorders. The aim of the present study has been to investigate differences between the four subtypes of the conversion disorder and to discuss the possibilities for a reclassification. Ninety-five patients were seen by two researchers and diagnosed as conversion disorders. The subtypes were determined according to DSM-IV criteria. All completed the Patients Information Form, developed by the researchers, and the Dissociative Experience Scale (DES). Twenty-four (25.2%) of the patients had motor symptoms or deficits (Type 1), 5 (5.2%) sensory symptoms or deficits (Type 2), 23 (24.2%) seizures or convulsions (Type 3) and 43 (47.3%) had mixed presentations (Type 4). There were statistically significant differences between the subtypes as concerns occupation, family history of psychiatric disorders, hospitalizations and place of settlement. Furthermore, the DES scores were statistically different between the groups of patients with different subtypes of conversion disorder. PMID- 14522605 TI - The Danish national multicenter schizophrenia project: assessment of psychotic patients for dynamic psychotherapy (APPP). AB - Clinicians need guidance in selecting schizophrenic patients for individual psychodynamic psychotherapy. The purpose of the present study was to investigate whether a 17-item questionnaire (APPP) for quantification of suitability for psychotherapy could predict continuance, adherence and outcome from individual dynamic psychotherapy with patients with schizophrenic psychoses. Therapists rated 72 first-episode patients with schizophrenic psychoses in the initial interview. The scores were then correlated to the course of therapy and change in symptoms. APPP was tested to have good psychometric characteristics and had a satisfactory interrater reliability ((ICC,2)=0.70), when four raters scored eight videotaped interviews with eight different patients. The scale scores were highly correlated to "therapeutic alliance" measured by the Working Alliance Inventory (WAI) and the Active Engagement Scale (AES). APPP suitability gave a good early prediction of adherence to therapy (continued in therapy at least 12 months and a satisfactory number of sessions), but had no predictive value of symptomatic improvement or improved social function after 1 year. PMID- 14522606 TI - Parent-child discrepancy in reporting children's post-traumatic stress reactions after a traffic accident. AB - This study examines possible parent-child discrepancies in the reporting of post traumatic stress reactions in children after a traffic accident. Sixteen children exposed to the same traffic accident were interviewed about post-traumatic stress reactions at 5 weeks and at 6 months after the event, utilizing the Child Posttraumatic Stress Reaction Index (CPTS-RI). Independently, the parents' reported their child's degree of post-traumatic stress reactions on the CPTS-RI: Parent questionnaire, at the same two times. Clinicians also assessed the children's level of general functioning on the Children's Global Assessment Scale. The children reported significantly more post-traumatic stress reactions than observed by their parents 4 weeks after the accident. The parent-child discrepancy was more pronounced among younger children. The level of children's self-reported post-traumatic stress reactions decreased significantly from the first to the second assessment. At the second assessment, 6 months after the accident, there was no significant parent-child discrepancy observed. The children showed a normal level of functioning despite their post-traumatic stress reactions. The reported parent-child discrepancy indicates that information about children's post-traumatic stress reactions after an accident is best obtained directly from the children. PMID- 14522607 TI - Traumatic injuries of children and adolescents--are they associated with psychiatric contacts? AB - Our aim was to investigate whether a large number of hospital visits by children and adolescents because of injuries are associated with psychiatric treatments and subsequent suicides. We examined the case records of 250 randomly chosen patients, 156 (62%) boys and 94 (38%) girls, out of 2306 outpatients who were 0 16 years old and had been treated because of traumatic injuries in Oulu University Hospital in 1984 and were alive in 1997. Boys with seven or more accidents had had psychiatric treatments more commonly than did boys with fewer accidents (39% vs. 8%). In addition, the case records of the patients who had died before the end of 1997 out of the 2306 patients were examined. Twenty-one (0.9%) patients (20 males and one female) had died, and seven (0.3%) of them had committed suicide. The traumatically injured male patients had a twofold suicide rate compared to the national average. The researchers also re-coded the causes of death from the death certificates. There seems to be a tendency to interpret adolescent suicides as accidental, as one of the seven registered suicides (14%), but six of the seven re-coded suicides (86%) had occurred before the age of 20. PMID- 14522608 TI - Quantitative electroencephalogram (qEEG) in combat veterans with post-traumatic stress disorder (PTSD). AB - Only a small number of studies have used quantitative electroencephalography (qEEG) in research of the post-traumatic stress disorder (PTSD). The results are not consistent. The aim of the present investigation was to compare qEEG in combat veterans with and without PTSD. The hypothesis is that differences among qEEG characteristics will be found regarding the presence/absence of PTSD. Seventy-nine combat veterans with PTSD comprised the experimental group and 37 veterans without PTSD were included as controls. After the informed consent, they were investigated by the resting EEG recordings. The results demonstrate that PTSD veterans had decreased alpha power and increased beta power. These results suggest an altered neurobiology in PTSD. Various explanations have been offered for alpha activity decrease observed in PTSD veterans. Increased beta rhythm may play a role as a potential marker in differentiating subtypes of PTSD. PMID- 14522609 TI - Validity of the Swedish version of the Strengths and Difficulties Questionnaire (SDQ-Swe). AB - The Strengths and Difficulties Questionnaire (SDQ) is an internationally widely used, brief screening instrument for mental health problems in children and teenagers. The SDQ probes behaviours and psychological attributes reflecting the child's difficulties as well as strengths, and targets hyperactivity/inattention, emotional symptoms, conduct problems, peer problems and prosocial behaviour. Also, the instrument taps the impact aspect, i.e. whether the child is judged to suffer from emotional or behavioural problems severe enough to cause distress or social impairment. Studies of the original English SDQ, as well as of translations into several other languages, attest to a compelling usefulness and validity of the instrument. In this investigation, the adequacy of the Swedish adaptation of the SDQ (SDQ-Swe) was tested in comparisons between parent reports on 5-15-year-old children drawn from a community sample (n=263) and from a child psychiatric sample (n=230). Results showed that the instrument differentiated well between the community and the psychiatric samples, the latter displaying more symptoms, fewer strengths and more social impairment. Moreover, ROC analyses showed satisfactory sensitivity and specificity of the principal scales of the SDQ-Swe at proposed cut-offs. Hence, results showed adequate validity of the SDQ Swe, suggesting that this new instrument, an instrument in tune with the ideas of contemporary child psychiatry and psychology, is a useful tool for mental health screening in children and adolescents. PMID- 14522610 TI - The roles of knowledge and supervision in work with suicidal patients. AB - Despite caregivers' impact on suicidal patients' compliance with treatment and suicide prevention, little is known about mental-health professionals' perceptions of work with suicidal patients. The roles of psychiatric staff's training and supervision in the care of suicidal patients were investigated by means of a postal questionnaire sent to a random sample of 1543 psychiatric staff members. The response rates were 71% for psychiatrists and 57% for nurses and assistant nurses. The responses of 53 psychiatrists, 164 nurses and 333 assistant nurses working with suicidal patients on a regular basis were compared and analysed using the Kruskal-Wallis test. Thirty-five per cent of the assistant nurses, 43% of the nurses and 74% of the psychiatrists who worked with suicidal patients on a regular basis perceived that they were sufficiently trained for this work, while 75% of the assistant nurses, 72% of the nurses and 34% of the psychiatrists received supervision in their work with suicidal patients. In spite of receiving supervision, nursing staff who perceived that they lacked training reported uncertainties in their work with suicidal patients to a larger extent than those who perceived that their training was sufficient. Uncertainties were significantly more prevalent among nursing staff than among psychiatrists. Basic and specific training in suicidology is needed and cannot be replaced solely by supervision, since psychiatric staff often have to deal with suicidal patients in emergency situations and must be able to rely on their own skills. PMID- 14522611 TI - Risk factors for drug addiction and its outcome. A follow-up study over 25 years. AB - The aim of the study was to investigate factors related to initiation and abandonment of illegal drugs. A follow-up study on 53 drug addicts originally hospitalized because of parenterally acquired hepatitis was performed using questionnaires. The majority stated that curiosity and peer pressure were the main reasons for starting drug use. Family conflicts, school and mental problems were each reported by about 40% of the subjects. Thirty-nine patients (74%) had abandoned addictive drugs during the approximately 25 years that had elapsed since the hospital stay. All of these thought that own efforts had been most important in this achievement. Other important factors were help and support from family and friends, establishing a family or experiencing acute hepatitis or an overdose. PMID- 14522612 TI - Dream screen phenomenon in psychotherapy and artistic work. AB - The aim of this article is to offer a point on the meaning of the dream screen phenomenon in psychotherapy and artistic work. Some current theories of early ego development, the formation of the dream screen and dream imagery seem to match the theories of symbol formation, artistic and literary creativity. In actual dreaming, the dream screen can exist in the form of empty sheets of paper, but also in the form of a landscape, hills or mountains. The activation of the dream screen is presented in the light of the psychotherapy of two depressed patients. Dreams implying activation of the dream screen have been interpreted to signify a turning point during psychotherapy. Dream screen can be considered as a phenomenon with different elements. Examples of dream screen symbols in Aleksis Kivi's poems and in Paul Cezanne's paintings are presented. PMID- 14522620 TI - Physicians underutilize topical retinoids in the management of acne vulgaris: analysis of U.S. National Practice Data. AB - BACKGROUND: Topical retinoids are effective in the treatment of acne, yet no current guidelines existed in the literature until recently recommending their use as an integral part of mild to moderate acne treatment. OBJECTIVE: To assess whether underutilization of topical retinoids occurs in clinical practice. DESIGN: Weighted data on representative office visits with acne vulgaris diagnoses were analyzed using the National Ambulatory Medical Care Survey (NAMCS), from 1990 to 1999. RESULTS: Of the 54.2 million acne vulgaris visits, retinoids were prescribed at 35.3% visits. Dermatologists prescribed significantly more retinoids than non-dermatologists (39.4%' versus 23%). Dermatology visit was a significant predictor of topical retinoid prescription (RR: 2.27,95% CI: 1.63-3.17). Data suggest that prescribing retinoids for acne has been increasing over the past decade, primarily among dermatologists. CONCLUSION: There is a major difference between suggested treatment guidelines for acne and actual practice in the community. There is distinct under utilization of topical retinoids for treatment of acne vulgaris by dermatologists and non-dermatologists. PMID- 14522621 TI - [Development of calcineurin blocking non-steroid topical immunosuppressants for effective management of eczema]. PMID- 14522622 TI - An open-label pilot study to evaluate the safety and efficacy of topically applied tacrolimus ointment for the treatment of hand and/or foot eczema. AB - BACKGROUND: Hand and foot eczema is a chronic skin disorder. Although topical corticosteroids are often used to control the predominant symptoms of the disease, the chronicity of the condition increases the risk of long-term adverse effects. A safer alternative is needed. OBJECTIVE: To evaluate the safety and efficacy of tacrolimus ointment 0.1% in hand and/or foot eczema. METHODS: Twenty five adults applied tacrolimus ointment 0.1% to affected areas three times daily for 8 weeks and were followed for 2 additional weeks. RESULTS: Except for vesiculation, compared with baseline there were significant improvements in erythema, scaling, induration, fissuring, composite severity, and pruritus (p<0.007). Two weeks after discontinuing treatment, significant improvement in scaling and composite severity (p<0.03) persisted, whereas erythema, induration, vesiculation, fissuring, and pruritus had returned to pre-treatment levels. CONCLUSION: Tacrolimus ointment 0.1% is a promising corticosteroid alternative for hand/foot eczema. PMID- 14522624 TI - Short-term growth and bone turnover in children undergoing occlusive steroid ('Wet-Wrap') dressings for treatment of atopic eczema. AB - To assess the effects of steroid wet-wrap therapy on short-term growth and bone turnover, eight prepubertal (M:F,5:3) children with a median age of 5.1years (range 3.3-8.8) were studied over a 2-week period prior to therapy and at 2-week intervals during therapy. Short-term growth was assessed by measuring lower leg length velocity (LLLV) by knemometry and bone and collagen turnover was assessed by urinary deoxypyridinoline crosslink excretion corrected for creatinine excretion (DPD). Median duration of study during occlusive dressings was 12 weeks (range 2-18). Topical beclomethasone dipropionate diluted 1:10 or 1:4 in white soft paraffin was applied under tubular (Tubifast) bandages in 7/8 children. Median LLLV before and during therapy were 0.43 mm/week (10(th),90(th) centile; 0.0,0.7) and 0.42 mm/week (10(th),90(th) centile; -0.35,1.01), respectively (not significant). Median DPD before and during therapy were 25.9 nmol/l/creatinine (10(th),90(th) centile; 20.8, 33.0) and 26.3 nmol/l/creatinine (10(th),90(th) centile; 21.7, 34.1) respectively (not significant). Non-invasive assessment of the effects of steroid wet-wrap therapy can be performed in children with eczema. These preliminary results show no substantial growth promoting or adverse effects of therapy. PMID- 14522623 TI - Acute and maintenance treatment of atopic dermatitis in children - two comparative studies with fluticasone propionate (0.05%) cream. AB - BACKGROUND: Two multicentre, randomised, parallel group, double-blind, comparative studies in children (2-14 yr) evaluated fluticasone propionate (FP) 0.05% cream for both acute and maintenance treatment of moderate to severe atopic dermatitis (AD). METHODS: One study compared FP with hydrocortisone (HC) 1% cream (FP 70, HC 67) and the other with hydrocortisone butyrate (HCB) 0.1% cream (FP 67, HCB 62). Treatments were applied twice daily, for 2-4 weeks until the AD was stabilised, and thereafter intermittently ('as required') for up to 12 weeks. RESULTS: The primary outcome measure, Total AD Score, recorded at the end of the acute and maintenance phases, was significantly lower (indicating improvements in disease severity) following treatment with FP compared with either HC or HCB (acute phase difference vs. HC, -2.39, 95%CI -3.47, -1.31; p<0.001 and vs. HCB, 1.25, 95%CI -2.46, -0.05; p=0.042) and (maintenance phase difference vs. HC, 1.88, 95%CI -3.20, -0.56; p=0.006 and vs. HCB, -1.39, 95%CI -2.72, -0.05; p=0.042). In both studies treatments were equally well tolerated with no visible signs of skin atrophy. CONCLUSION: In both the acute and longer term management of AD in children, FP demonstrated a high level of efficacy and maintenance of disease control with a tolerability similar to HC 1%, a lower potency corticosteroid. PMID- 14522625 TI - The treatment of atopic dermatitis with licorice gel. AB - Glycyrrhiza glabra L. has been used in herbal medicine for skin eruptions, including dermatitis, eczema, pruritus and cysts. The effect of licorice extract as topical preparation was evaluated on atopic dermatitis. The plant was collected and extracted by percolation with suitable solvent. The extract was standardized, based on Glycyrrhizinic acid by using a titrimetry method. Different topical gels were formulated by using different co-solvents. After standardizing of topical preparations, the best formulations (1% and 2%) were studied in a double-blind clinical trial in comparison with base gel on atopic dermatitis over two weeks (30 patients in each group). Propylene glycol was the best co-solvent for the extract and Carbopol 940 as gelling agent showed the best results in final formulations. The quantity of glycyrrhizinic acid was determined 20.3% in the extract and 19.6% in the topical preparation. Two percent licorice topical gel was more effective than 1% in reducing the scores for erythema, oedema and itching over two weeks (p<0.05). The results showed that licorice extract could be considered as an effective agent for treatment of atopic dermatitis. PMID- 14522626 TI - Psoriasis disease severity measures: comparing efficacy of treatments for severe psoriasis. AB - Measurement of psoriasis disease severity and effectiveness of treatment involves both objective and subjective assessments.1 Comparing the efficacy of different treatments is complicated by the use of different metrics for measuring outcomes.2 Because these measures are not used routinely in clinical practice, interpreting these data, in particular assessing the degree of clinically meaningful improvement, is difficult. The drug approval process and product labeling reflect historical changes in standards of efficacy measurement.3 This paper reviews the metrics used to evaluate psoriasis treatment and compares available information on approved treatments for severe psoriasis. It further attempts to elucidate the value of these metrics and provide some guidance in properly evaluating the relative efficacy of current proven therapy with new treatments. While clinical trials are somewhat artificial, they provide proof that a drug is more effective than placebo. Efficacy in clinical practice, however, may be very different from the clinical trial setting. Comparison of efficacy under the current circumstances of varying evaluative metrics scales is possible with proper knowledge of the functionality of these methods. PMID- 14522627 TI - Single-blind and comparative clinical study of the efficacy and safety of benzoyl peroxide 4% gel (BID) and adapalene 0.1% Gel (QD) in the treatment of acne vulgaris for 11 weeks. AB - objectives: To compare the efficacy and tolerability of a gel containing benzoyl peroxide 4%, used twice daily, with a gel containing adapalene 0.1% used once daily, in the treatment of acne vulgaris for 11 weeks. methods: 178 patients bearing acne vulgaris, aged between 13 and 30 years, were studied in a comparative and single-blind clinical study. The 178 patients were divided into two groups: 89 patients treated with benzoyl peroxide 4% and 89 patients treated with adapalene 0.1%. The treatment duration was 11 weeks. The efficacy assessment was conducted through an accounting of both the inflammatory and non-inflammatory lesions in all visits. The safety assessment was conducted through reports regarding adverse reactions and local tolerance in all visits and an overall tolerance at the end of the study. conclusions: The results showed that both treatments were efficient in the reduction of acne lesions after 11 weeks treatment and were well tolerated, without any serious adverse event report. The benzoyl peroxide 4% was superior in the reduction of the number of inflammatory and non-inflammatory lesions at weeks 2 and 5, when compared to adapalene 0.1%. PMID- 14522628 TI - Skin rash and splinter hemorrhages from ganciclovir. AB - Ganciclovir is a nucleotide-analogue similar to acyclovir, which has an in vitro activity against herpes simplex type 1, herpes simplex type 2 and varicella zoster virus. Numerous studies suggest that ganciclovir has clinical efficacy against cytomegalovirus disease, as well as an in vivo antiviral effect, and that this agent reduces morbidity of serious cytomegalovirus infections in immunocompromised patients. Generalised cutaneous rash associated with ganciclovir therapy has rarely been reported in literature. PMID- 14522629 TI - Henoch-Schonlein purpura following hepatitis B vaccination. AB - The use of recombinant hepatitis B vaccines has led to the effective prevention of hepatitis B infection and its chronic sequelae in immunocompetent individuals. Whilst rare, a variety of serious adverse effects have been reported following vaccination including cutaneous vasculitis in eight previous cases. We describe a case of Henoch-Schonlein purpura developing after hepatitis B vaccination. PMID- 14522630 TI - Discoid lupus erythematosus treated with cryotherapy. AB - A female is presented with multiple longstanding hypertrophic discoid lupus erythematosus lesions resistant to topical and systemic therapy for years. She suffered severe itching and pain localized to the lesions. Treatment with cryosurgery resulted in complete healing of the lesions, leaving slightly hypopigmented soft scars. There was no tendency of relapse during a follow-up period of ten years. PMID- 14522631 TI - Epidermodysplasia verruciformis with multiple mucosal carcinomas treated with pegylated interferon alfa and acitretin. AB - Epidermodysplasia verruciformis (EV) is characterized by abnormal genetically determined susceptibility to widespread and persistent infection of the skin with human papillomaviruses (HPV). The infection results in disseminated pityriasis versicolor-like lesions and flat warts. Skin malignant changes are very common and occur on sun-exposed areas. Several treatments have been used but without consistent benefit. Recently, retinoids and alpha-interferon, alone or in combination, have been reported to be of value in the therapy of EV lesions. We present the case of a 43-year-old white female affected by EV who developed multiple squamous cell carcinomas in the oral and genital mucosae during the previous four years. Both wart and cancer lesions harbored HPV24 along with the novel putative HPV type FA51. The patient was treated with a combination of acitretin (0.2 mg/kg per day) and peginterferon alfa-2b (1 microg/kg per week s.c.) for one year, with marked improvement of verrucous lesions and no recurrence of mucosal cancer. Thereafter, interferon was stopped whereas acitretin therapy was continued, but a new Bowen's disease developed in the perianal region, and the acitretin dose was increased at 0.5 mg/kg per day. At six-month follow-up, only a low number of flat warts persisted, and no clinical signs of cutaneous or mucosal carcinoma were evident. PMID- 14522632 TI - Erythema multiforme-like eruption as a result of 'Solaraze' treatment. PMID- 14522633 TI - Severe pruritus in a patient with urticaria pigmentosa treated with topical 5% urea and 3% polidocanol cream. PMID- 14522634 TI - Atopic dermatitis management with tacrolimus ointment (Protopic). AB - Tacrolimus ointment is the first of a new class of non-steroidal topical immunomodulators indicated for the treatment of atopic dermatitis. Topical tacrolimus has been subject to an extensive clinical development program involving more than 16,000 patients. A clinical trial program, including vehicle controlled studies, short- and long-term comparative studies and long-term safety studies, has investigated tacrolimus 0.1% and 0.03% ointment for the treatment of atopic dermatitis in adults and children aged 24 months and older. Tacrolimus monotherapy is rapidly effective, resulting in clinical improvements within three days of starting therapy, and produces a progressive increase in efficacy that is sustained during long-term treatment. Tacrolimus treats the signs and symptoms of atopic dermatitis, reduces the incidence of flares, and offers the potential for long-term disease control. No major safety concerns have been reported to date. Tacrolimus ointment is generally well tolerated, the primary adverse events being mild to moderate and transient application-site reactions: skin burning, pruritus and erythema. Tacrolimus ointment is a significant advance in dermatology and provides physicians with an alternative to conventional topical corticosteroid therapy. PMID- 14522635 TI - Current treatment patterns in non-melanoma skin cancer across Europe. AB - With marked increases in the annual incidence of non-melanoma skin cancer (NMSC) across the globe, its management is of increasing concern to dermatologists. This paper summarises the epidemiology and risk factors and provides an overview of treatment approaches in NMSC across Europe, including surgery, topical 5 fluorouracil and cryotherapy, in the context of the trade-offs that exist in finding optimal treatment outcomes. The paper will also briefly examine new approaches such as immunomodulators and the growing body of data on photodynamic therapy (PDT) using methyl aminolevulinate (MAL), including the authors' personal experience of the efficacy and cosmetic results obtained with these newer therapies. PMID- 14522636 TI - The role of light in the treatment of non-melanoma skin cancer using methyl aminolevulinate. AB - Photodynamic therapy (PDT) is a developing approach to the treatment of cancer and other diseases that involves the use of light to activate photosensitizer molecules. The light energy absorbed by the photosensitizer is transferred to molecular oxygen, which is converted into the highly reactive and cytotoxic species, singlet oxygen. Topical agents such as aminolevulinic acid (ALA) or methyl aminolaevulinate (MAL) may be used for PDT of non-melanoma skin cancers (NMSC) because, in vivo, these agents stimulate the production of porphyrins which act as powerful photosensitisers. This brief review focuses on the use of light to activate MAL, which is now an approved drug (Metvix for certain NMSCs in the European Union. Porphyrins produced by the action of MAL can be activated using red light, which is also capable of deeply penetrating the skin. A number of light sources are available for treatment of NMSC using MAL, including very convenient non-laser sources such as non-coherent filtered lamps and, more recently, sources containing arrays of light-emitting diodes (LEDs). The Aktilite lamp, specifically designed for use with Metvix cream, has an emission spectrum that closely matches the red light absorption profile of PpIX. PMID- 14522637 TI - Clinical efficacy of methyl aminolevulinate (Metvix) photodynamic therapy. AB - Several studies have reported the efficacy of photodynamic therapy (PDT) in the treatment of non-melanoma skin cancer (NMSC) and pre-cursor lesions, such as actinic keratosis (AK). Recent studies investigating the use of methyl aminolevulinate (MAL, Metvix PDT have made comparisons to existing standard treatments for basal cell carcinoma (BCC) and AK in terms of efficacy, adverse events and cosmetic outcome. This review considers these studies as a body of evidence that supports the use of MAL PDT over traditional therapies in these conditions. The data thus far show PDT to be superior to cryotherapy, 5 fluorouracil (5FU) and excisional surgery in terms of cosmetic outcome, and equivalent in efficacy to other treatment modalities in managing AK and BCC. The studies also show an advantage in the treatment of extensive lesions. PMID- 14522638 TI - Methyl aminolevulinate (Metvix) photodynamic therapy - practical pearls. AB - Topical photodynamic therapy (PDT) is an effective treatment for certain non melanoma skin cancers (NMSCs), including superficial and nodular basal cell carcinomas (sBCC and nBCC), actinic keratosis (AK) and Bowen's disease. Methyl aminolevulinate (MAL, Metvix) is licensed in Europe for use in PDT for sBCC, nBCC and thin or non-hyperkeratotic and non-pigmented AK on the face and scalp, where other therapies are unsuitable. Optimal PDT response can be achieved through appropriate patient selection and lesion preparation. Evidence of efficacy is reviewed from guidelines and clinical experience. Red light from an LED source offers a relatively efficient method of activating the photodynamic reaction. The most common side effect of PDT is pain, burning or stinging discomfort at the site of treatment, although most patients do not request pain relief. The incidental observation of surface fluorescence three hours after photosensitizer application can be utilized for tumour detection as well as delineation. Topical PDT using Metvix MAL offers a practical non-invasive therapy option with the potential for high efficacy and good cosmesis. PMID- 14522639 TI - Preventing alcohol crashes: the role of ignition interlocks. PMID- 14522640 TI - International symposium on enhancing the effectiveness of alcohol ignition interlock programs. PMID- 14522641 TI - Barriers to interlock implementation. AB - This commentary reviews current and past vehicle interlock programs for impaired driving offenders with a focus on the challenges that courts and motor vehicle departments face in attempting to implement mandatory programs as required by current federal legislation. There are few offenders in interlock programs compared to the large number of impaired drivers arrested each year. This suggests that, to increase participation, courts will have to threaten more severe sanctions for those offenders who reject interlock programs. A combination of electronic house arrest with interlock programs is suggested as a method of maximizing interlock use. PMID- 14522642 TI - The alcohol interlock: an underutilized resource for predicting and controlling drunk drivers. AB - This report summarizes evidence presented during the Third Annual Ignition Interlock Symposium at Vero Beach, Florida, 29 October 2002. The ignition interlock prevents a car from starting when blood alcohol concentration (BAC) is elevated. We review some of our prior work as well as introduce previously unpublished results to demonstrate the manner in which the data recorded by the alcohol ignition interlock device can serve as an advance predictor of future driving under the influence (DUI) of alcohol risks. Data used in this current report represent approximately 2,200 ignition interlock users from Alberta, Canada, and about 8,000 interlock users from Quebec, Canada; the Alberta data set contained 5.5 million breath tests and the Quebec data 18.8 million breath tests. All tests are time and date stamped and this information was used to characterize patterns of BAC and vehicle use, and the relationship between BAC elevations and DUI offenses that accumulated after the interlock was removed from the vehicles. Findings from Cox regression show that BAC elevations >.02-.04% are more potent predictors of repeat DUI (p<.0001) than even prior DUI (p<.006), usually found to be the strongest indicator of driver risk. Prior DUI obviously has no use for scaling the risk of first-time offenders. Drivers who are both multiple offenders and who have more than a few elevated interlock BAC tests are much more likely to repeat DUI. The timing and pattern of elevated BAC tests provided during the time drivers were required to use an alcohol ignition interlock device are remarkably similar on both a daily basis and an hourly basis when the interlock programs from the two provinces are compared directly. Both provinces had higher rates of elevated tests on Saturday and Sunday, and the fewest elevated tests on Tuesdays. The absolute rate of elevated tests is similar despite the two provinces adhering to different interlock lockout points (.02% Quebec;.04% Alberta). Charts tracking the Monday-Friday timing of elevated BAC tests by hour are nearly identical for both provinces. The most elevated BAC tests occurred between 7 and 9 A.M. Monday to Friday, even though most vehicle start attempts occurred much later in the day. This higher rate of elevated morning BAC likely represents drinking from the prior evening with alcohol not yet cleared from circulation; those with elevated BAC in the early morning were more likely to have a repeat offense even after accounting for prior DUI and the higher overall rate of elevated BAC tests. This is viewed as evidence of a drinking problem that will lead to impaired driving after the controlling function of the interlock is removed. Policy changes are discussed that might take better advantage of interlock information to improve the public response to drunk driving. PMID- 14522643 TI - The impact of mandatory versus voluntary participation in the Alberta ignition interlock program. AB - Research has demonstrated that participation in an interlock program significantly reduces the likelihood of subsequent driving while intoxicated (DWI) convictions at least so long as the interlock device is installed in the vehicle. Despite the growing number of jurisdictions that allow interlock programs and the demonstrated success of these programs, the proportion of DWI offenders who actually have the device installed is minimal. In an effort to increase the proportion of offenders using interlocks, some jurisdictions require offenders to install an interlock as a condition of license reinstatement whereas others merely offer offenders a reduction in the period of hard suspension if they voluntarily participate in an interlock program. The objective of the present study was to determine the extent to which voluntary interlock participants are more or less successful in terms of subsequent recidivism than those for whom interlock program participation has been mandated. The issue was addressed using data from the interlock program in Alberta, Canada, which provides for both mandatory and voluntary participation. The recidivism experience of voluntary and mandatory interlock participants was examined both during and after the period of interlock installation. Cox regression revealed that, after controlling for (or equating) the number of prior DWI offenses, the survival rates of DWI offenders who were ordered to participate in the interlock program did not differ from those of voluntary participants. These results suggest that further use of mandatory interlock programs should be just as successful as voluntary programs when offenders share characteristics with those studied in Alberta. PMID- 14522644 TI - Breath alcohol ignition interlock devices: controlling the recidivist. AB - This study compares the recidivism rates of two groups of Illinois drivers who had their driver's licenses revoked for alcohol-impaired driving and who received restricted driving permits. Drivers in both groups had more than two driving under the influence (DUI) actions against their record within 5 years or were classed as level III alcohol dependents. Drivers in one group were required to install breath alcohol ignition interlock devices in their vehicles and drivers in the other group were not. The research found that drivers with the interlock were one-fifth as likely to be arrested for DUI during the 1 year the device was installed as the comparison group, which did not have the device. However, once the ignition interlock was removed, drivers in this group rapidly returned to DUI arrest rates similar to those in the comparison group. These findings echo previous literature. Additionally, the study showed that this voluntary program in Illinois reached only 16% of the drivers who met the requirements for installing the interlock device. Finally, this study found that individuals who were removed from the interlock program and returned to revoked status continued to drive. Within 3 years, approximately 50% of this latter group were involved in a crash or were arrested for DUI or with an invalid driver's license. Conclusions drawn from the study suggest that the breath alcohol ignition interlock device is effective in preventing continued driving while impaired. However, the large scale effectiveness of the device is limited since most of the drivers eligible for the device do not have it installed. To have a significant impact, the interlock device must represent a better alternative to drivers whose licenses were suspended or revoked because of alcohol arrests compared to remaining on revoked status without having the device installed. Finally the research suggests that, given the rapid return to predevice recidivism, the devices should remain installed until drivers can demonstrate an extended period of being alcohol free. PMID- 14522645 TI - Prediction of cervical spine injury risk for the 6-year-old child in frontal crashes. AB - This article presents a series of 49 km/h sled tests using the Hybrid III 6-year old dummy in a high-back booster, a low-back booster, and a three-point belt. Although a 10-year review at a level I trauma center showed that noncontact cervical spine injuries are rare in correctly restrained booster-age children, dummy neck loads exceeded published injury thresholds in all tests. The dummy underwent extreme neck flexion during the test, causing full-face contact with the dummy's chest. These dummy kinematics were compared to the kinematics of a 12 year-old cadaver tested in a similar impact environment. The cadaver test showed neck flexion, but also significant thoracic spinal flexion which was nonexistent in the dummy. This comparison was expanded using MADYMO simulations in which the thoracic spinal stiffness of the dummy model was decreased to give a more biofidelic kinematic response. We conclude that the stiff thoracic spine of the dummy results in high neck forces and moments that are not representative of the true injury potential. PMID- 14522646 TI - Seat properties affecting neck responses in rear crashes: a reason why whiplash has increased. AB - Whiplash has increased over the past two decades. This study compares occupant dynamics with three different seat types (two yielding and one stiff) in rear crashes. Responses up to head restraint contact are used to describe possible reasons for the increase in whiplash as seat stiffness increased in the 1980s and 1990s. Three exemplar seats were defined by seat stiffness (k) and frame rotation stiffness (j) under occupant load. The stiff seat had k=40 kN/m and j=1.8 degrees /kN representing a foreign benchmark. One yielding seat had k=20 kN/m and j=1.4 degrees /kN simulating a high-retention seat. The other had k=20 kN/m and j=3.4 degrees /kN simulating a typical yielding seat of the 1980s and 1990s. Constant vehicle acceleration for 100 ms gave delta-V of 6, 10, 16, 24, and 35 km/h. The one-dimensional model included a torso mass loading the seatback, head motion through a flexible neck, and head restraint drop and rearward displacement with seatback rotation. Neck displacement was greatest with the stiff seat due to higher loads on the torso. It peaked at 10 km/h rear delta-V and was lower in higher-severity crashes. It averaged 32% more than neck displacements with the 1980s yielding seat. The high-retention seat had 67% lower neck displacements than the stiff seat because of yielding into the seatback, earlier head restraint contact and less seatback rotation, which involved 16 mm drop in head restraint height due to seatback rotation in the 16 km/h rear delta-V. This was significantly lower than 47 mm with the foreign benchmark and 73 mm with the 1980s yielding seat. Early in the crash, neck responses are proportional to ky/mT, seat stiffness times vehicle displacement divided by torso mass, so neck responses increase with seat stiffness. The trend toward stiffer seats increased neck responses over the yielding seats of the 1980s and 1990s, which offers one explanation for the increase in whiplash over the past two decades. This is a result of not enough seat suspension compliance as stronger seat frames were introduced. As seat stiffness has increased, so have neck displacements and the Neck Injury Criterion (NIC). High-retention seats reduce neck biomechanical responses by allowing the occupant to displace into the seatback at relatively low torso loads until head restraint contact and then transferring crash energy. High-retention seats resolve the historic debate between stiff (rigid) and yielding seats by providing both a strong frame (low j) for occupant retention and yielding suspension (low k) to reduce whiplash. PMID- 14522647 TI - Seat influences on female neck responses in rear crashes: a reason why women have higher whiplash rates. AB - Since the earliest crash investigations, whiplash has been found to occur more often in women than men. This study addresses seat properties that may explain a reason for the higher rates in women, and changes in whiplash in general over the past two decades. Three exemplar seats were defined on the basis of seat stiffness (k) and frame rotation stiffness (j) for rearward occupant load. Stiff seats have k=40 kN/m and j=1.8 degrees /kN representing a foreign benchmark loaded by a male. One yielding seat had k=20 kN/m and j=1.4 degrees /kN simulating a high-retention seat (1997 Grand Prix) and another k=20 kN/m and j=3.4 degrees /kN simulating a 1980s to 1990s yielding seat (1990 Buick Park Avenue). Constant vehicle acceleration for 100 msec gave delta-V of 6, 10, 16, and 24 km/h. The one-dimensional model included a torso mass loading the seatback with flexible neck and head mass. Based on biomechanical data and scaling, neck stiffness was 5 kN/m and 3 kN/m for the male and female, respectively. Based on validation tests, seat stiffness was 25% less with the female. Occupant dynamics were simulated in a step-forward solution based on the differential displacement between the head, torso, and seat up to head restraint contact. Neck responses were 30% higher in the female than male through most of the rear impact and are proportional to (kF/mTF)/(kM/mTM), which is the ratio of seat stiffness divided by torso mass for the female and male. Neck displacements were higher with the stiff seat than the 1990 C car seat for both the female and male. They peaked at 10 km/h and dropped off for higher severity crashes due to the shorter time to head contact. Neck displacements were greater in the female than male for the lowest severity crashes with the stiff and 1990 C car seats, when displacement was scaled for equal tolerance. The female in 1997 W car seat had the lowest neck displacements. Stiff seats increased neck displacements over the yielding seats of the 1980s in rear crashes. The trend is similar in men and women, but early neck displacements are greater in women because of a higher ratio of seat stiffness to torso mass. This implies that seat stiffness is not sufficiently low in proportion to the female mass in comparison to males. The j and k seat properties influence neck biomechanics and occupant dynamics, but k is important in determining early response differences between males and females. PMID- 14522648 TI - A new laboratory rig for evaluating helmets subject to oblique impacts. AB - Current requirements and regulations governing motorcycle helmets around the world are based on test results of purely radial impacts, which are statistically rare in real accidents. This study presents a new impact rig for subjecting test helmets to oblique impacts, which therefore is able to test impacts of increased statistical relevance to real motorcycle accidents. A number of different head helmet interfaces have been investigated. A test rig was constructed to produce oblique impacts to helmets simulating those occurring in real motorcycle accidents. A Hybrid III dummy head was fitted with accelerometers to measure the accelerations arising during impact testing. The equipment used for data collection was validated in both translational and rotational acceleration. In order to better resemble the human head, an artificial scalp was fitted to the hybrid dummy. The same test rig was used to investigate the performance of a number of different helmets. Impact velocities ranging from 7.3 to 9.9 m/s were tested using a number of different impact angles and impact areas. This study shows that the new test rig can be used to provide useful data at speeds of up to 50 km/h and with impact angles varying from purely tangential to purely radial. The rotational accelerations observed differ greatly depending on both helmet and scalp designs. For example, a helmet with a sliding outer shell placed on an experimental head fitted with an artificial scalp (made to resemble the human scalp) reduces rotational accelerations of the head by up to 56%, compared with those of an experimental head fitted with a fixed scalp and conventional helmet. The degree of slippage between the skull and the scalp, and between the scalp and the helmet, leads to considerable variation in the results. This innovative test rig appears to provide an accurate method for measuring accelerations in an oblique impact to a helmet. In order to obtain a good level of repeatability in oblique impact testing, it is crucial that the helmet be fixed to the head in the exact same way in each individual test. Both the position and the angle of impact must be reproduced identically in each test. The test rig used here has shown that this type of rig can be used to compare different helmet designs, and it therefore is able to contribute to achieving safer helmets. PMID- 14522649 TI - Socioeconomic background and road traffic injuries: a study of young car drivers in Sweden. AB - The aim of this study is to explore the manner in which different measures of original socioeconomic position (SEP) influence road traffic injuries (RTIs) among young car drivers in Sweden. The study consists of young people age 16-23. Subjects were taken from the Swedish Population and Housing Census of 1990 (n=727,995), and followed up by a search for cases of injury to car drivers in Sweden's National Hospital Discharge Register over the years 1991-96 (n=1,599). Household SEP was measured using social class, education, and disposable income. Relative risks were estimated by Poisson regression and population attributable risks were computed for each measure of SEP. Children of unskilled workers, of the self-employed, and of farmers, as well as children of parents with compulsory education only showed an increased risk of injury as car drivers compared to children in the highest socioeconomic group and children of highly educated parents. By contrast, level of household disposable income was found not to vary with RTI among young drivers. Twenty-five percent of the injuries could be avoided if all young people had the injury rate of the highest socioeconomic group, and 29% if all young people had the injury rate of those with highly educated parents. The reduction of risk differences based on household SEP calls for consideration of factors related to both differential exposure and differential susceptibility, which may be addressed in driver education. PMID- 14522650 TI - Head and neck injuries in fatal motorcycle collisions as determined by detailed autopsy. AB - Detailed layer-by-layer autopsy of the head and neck was performed on a prospective series of 73 fatally injured motorcyclists in order to identify occult injuries, particularly soft tissue neck injuries such as hemorrhage of vertebral and carotid arteries. The fatal cases were gathered as part of a larger study of 1,082 on-scene in-depth motorcycle crash investigations in Thailand. Detailed neck dissection was done on nearly all fatal cases. Injuries were coded using the 1990 revision of the Abbreviated Injury Scale (AIS 90) and an Injury Severity Score (ISS) was determined for each case. Additional AIS codes are proposed for neck injuries that were often identified during the detailed autopsy procedures, but which are not listed explicitly among existing AIS codes. Helmet use was determined based on analysis of injury patterns and helmet damage with consideration also given to witness statements. Both helmeted and unhelmeted motorcyclists showed a high frequency of occult neck injuries such as hemorrhages in the carotid sheath or surrounding the vertebral arteries, phrenic nerve, or brachial plexus. These soft tissue neck injuries sometimes accompanied more obvious injuries to cervical vertebrae or spinal cord, but about one-third of riders had no obvious injury to suggest the presence of occult neck injury. Twenty-eight motorcyclists had been wearing a helmet at the start of the collision sequence, but only nine helmets remained in place through the entire collision event. Helmeted riders showed more severe somatic (below-the-neck) injuries than unhelmeted riders, suggesting helmeted riders are less likely to die in low-threat accidents with somatic injuries below AIS-3. The most significant finding of this study was the identification of serious internal neck injuries despite the absence of external physical evidence of trauma to the neck. Virtually all riders with significant head injuries showed some of these soft tissue neck injuries. Approximately one-third of the critically injured riders who survived at least a few hours before death showed serious occult soft tissue neck injuries. PMID- 14522651 TI - A preliminary analysis of aortic injuries in lateral impacts. AB - Injuries to the aorta are among the more serious injuries that result from vehicle impacts, and often may be fatal. This article examines the incidence of aortic injuries in the United States and United Kingdom by using two international databases of real-world crashes. The main outcome of interest was the level of risk associated with each principal direction of force for drivers and front-seat passengers with respect to sustaining aortic injuries. The results indicate that the risk of sustaining an injury to the aorta is greater for near side crashes than for far-side crashes. Further it is apparent that, given a near side crash, the risk of an aortic injury is greater on the left side of the body (and left side of the vehicle) than on the right. It also was found that the delta-V of crashes where occupants sustained an injury to the aorta was considerably higher than crashes where occupants did not sustain aortic injuries. It is speculated that the anatomical asymmetry of the thorax might play a role in the differences seen in injury risk associated with different impact directions. The results presented in this article could be of use to both the emergency physician treating patients involved in motor vehicle collisions as well as the engineer involved in occupant design countermeasures. Limitations and further planned research are discussed. PMID- 14522652 TI - Characterization of fatal occupational versus nonoccupational motor vehicle collisions in Kentucky (1998-2000). AB - Motor vehicle collisions (MVCs) are the leading cause of occupational fatalities in Kentucky as well as in the nation. The characteristics of and contributing factors for occupational versus nonoccupational MVC fatalities in the Commonwealth of Kentucky were examined from 1998 to 2000. Semi trucks were most frequently involved in fatal occupational MVCs, and passenger cars were most frequently involved in nonoccupational MVCs. More than half of the decedent drivers resided outside of Kentucky. The percentage of occupational fatalities occurring on a four-lane highway was double the percentage observed for nonoccupational MVC fatalities. In addition, an increased proportion of occupational MVC deaths occurred on limited access highways compared to nonoccupational fatalities. When human factors contributing to these fatal incidents were examined, the two primary human factors involved in occupational motor vehicle fatalities were driver distraction/inattention and falling asleep, whereas unsafe speed and alcohol were the primary human factors contributing to a nonoccupational fatality. These results suggest that semi drivers traveling on four-lane highways are more at risk for a fatal occupational injury in Kentucky. Therefore, additional epidemiological studies are needed to further examine human factors, the nature of the Kentucky highway system, and trucking controls (e.g., weigh station hours of operation) within the Kentucky transportation industry. PMID- 14522653 TI - Ethnicity as a factor in pizza delivery crashes. AB - A subjective assessment of patients admitted acutely to the orthopedic directorate revealed that there had been--over a short period of time--a small series of patients who had sustained fractures during the course of their employment as pizza delivery moped drivers. Consequently an observational study was performed in our district general hospital in order to investigate this further, and establish the demographic domain of the patients involved and the cost of their injuries. Our hospital has a patient catchment area of 350,000 and an emergency department that treats 95,000 patients annually. The patients in this series were described in terms of their injuries, ethnicity, and cost of injury treatment. The main findings were that pizza delivery personnel admitted acutely were male Afghans, with poor English language skills, who sustained significant fractures in moped accidents that were expensive to treat. PMID- 14522654 TI - Global priorities for vehicle safety. PMID- 14522655 TI - Booster seats for children: closing the gap between science and public policy in the United States. PMID- 14522657 TI - Effects of red light cameras on violations and crashes: a review of the international literature. AB - Red light running is a frequent cause of motor vehicle crashes and injuries. A primary countermeasure for red light running crashes is police traffic enforcement. In recent years, many police agencies have begun using automated red light cameras as a supplement to conventional enforcement methods. The present study reviewed and evaluated available evidence in the international literature regarding the effectiveness of cameras to reduce both red light violations and crashes. Camera enforcement generally reduces violations by an estimated 40-50%. In terms of crash effects, most studies contain methodological flaws that, to varying degrees, either overestimate (failure to adjust for regression to the mean) or underestimate (comparison with nearby signalized intersections affected by cameras) crash effects. Mindful of these limitations, the research generally indicates that camera enforcement can significantly reduce injury crashes at signalized intersections, in particular right-angle injury crashes. Most studies reported increases in rear-end crashes following camera installation. Taken together the studies indicate that, overall, injury crashes, including rear-end collisions, were reduced by 25-30% as a result of camera enforcement. PMID- 14522656 TI - An overview of research advances in road traffic trauma in China. AB - With rapid development of social economies, road traffic accidents (RTAs) have continued to increase, and have become the "primary public hazard" to humans. Road traffic trauma (RTT) is a major cause of death in young people of all motorized countries. This article reviews the current advances in RTT research, in order to find some approaches to improving traffic administration and reducing RTAs and RTT. All available data were collected from government, literature, our own research, and conference proceedings. Statistical analysis from every country showed that human factors were still the main cause of RTAs, accounting for more than 90%. Vehicle and road factors caused 3-5% and less than 2% of the total RTAs, respectively. Approximately 85% of RTAs were caused by 21 to 45-year-olds. About 50% of deaths due to RTAs in the United States were related to drinking. In China, RTAs due to driver drinking accounted for 0.29-1.48%. About 6-8% of drivers were prone to RTA, causing 30-40% of RTAs. Seat belts are an effective way to prevent casualties, reducing mortality and morbidity by 13-50%. In China, about 70% of RTAs were related to bicycles. Prehospital emergency treatment is very important. About 35% of deaths may be avoided if the injured receive early and effective treatment. From 1983 to 1992 the mortality of RTT increased by 13% in 18 developing countries, while it decreased by 18% in 13 developed countries, indicating the importance of comprehensive treatment of traffic administration. In addition many advances have been made in basic scientific research of RTT, such as development of serial bioimpact machines and investigation of biomechanical and biochemical mechanisms of impact injuries. In this century, RTAs and RTT are predicted to continue to increase in many countries, especially in developing ones. Full cooperation and comprehensive treatment should be performed in order to improve traffic safety. PMID- 14522658 TI - How future trends in societal aging, air bag availability, seat belt use, and fleet composition will affect serious injury risk and occurrence in the United States. AB - The objective of this study was to quantify the importance of societal aging relative to other factors that are known to affect injury risk. An aging population's effect on Maximum Abbreviated Injury Scale (MAIS) 3+ injury trends for passenger car drivers in frontal crashes was projected and compared to the effects of projected changes in fleet composition, changes in seat belt usage, and changes in air bag availability. It was determined that increased frailty due to an aging population will result in 19,816 to 43,568 additional injuries to passenger car drivers in frontal crashes from 1996 to 2012. Aging was shown to have an effect similar to the increased presence of light trucks in the fleet (a cumulative increase of approximately 47,428 injuries). Aging and changing fleet composition were shown to have a smaller effect than the projected increases in seat belt use or air bag availability, though the effect of increased belt use is not much greater than the effect of aging. After 2012, however, air bag availability and seat belt use will plateau, while societal aging will continue. PMID- 14522659 TI - The influence of the age and sex distributions of drivers on the reduction of impaired crashes: Ontario, 1974-1999. AB - Legislative changes and public media campaigns to prevent impaired driving are often cited as explanations for the reduction in the rate of impaired crashes over the past 25 years in most of the industrialized world. Other factors may have contributed to these reductions, such as changes in the age and sex distribution of the driver population. The primary purpose of this article is to assess the extent to which the reduction in impaired crashes in Ontario, Canada, may be attributable to the changing age and sex distribution of drivers. In Ontario, the rate of impaired crashes declined by 78.1% from 1974 to 1999. During this time period, the average age of drivers increased from 39.4 years in 1974 to 43.2 in 1999. Similarly, from 1974 to 1999 the percentage of all drivers that were women increased from 39.6% to 46.8%. Since statistics show the likelihood of impaired crashes is lower for both older drivers and women, the reduction of impaired crashes is partially due to these demographic changes. Using indirect standardization, the aging population accounted for an 8.6% decline in the rate of impaired crashes. The changing sex distribution of drivers accounted for a 9.4% decline in impaired crashes. Other global factors may also help to explain the reduction of impaired crashes, such as general road safety improvements and reductions in per adult consumption of alcohol. PMID- 14522660 TI - Reliability of police-reported information for determining crash and injury severity. AB - Posted speed limit and police-reported injury codes are commonly used by researchers to approximate vehicle impact and occupant injury severity. In-depth crash investigations, however, produce more precise measures of crash and injury severity: change in velocity (delta-V) for crash severity and Abbreviated Injury Scale (AIS) scores for injury severity. A comparison of data from police crash reports with that gathered by National Automotive Sampling System (NASS) investigators highlighted the inadequacy of speed limit and police injury codes as proxies for delta-V and AIS injury severity. In general, delta-V increased with speed limit and higher values of AIS were associated with higher police coded injury severity, but there were a number of anomalies. In particular, 49% of the drivers coded by police as having incapacitating injuries actually had sustained no more than minor injuries. This overstatement of injury severity was less frequent among male (44%) and elderly (37%) drivers than among female (53%) and nonelderly (50%) drivers. Also, 79% of the investigated vehicles that crashed on roads posted at 60 mph (96 km/h) or higher experienced a delta-V less than 25 mph (40 km/h). Safety studies depending on data from only police reports to establish injury or crash severity therefore could produce erroneous results. PMID- 14522661 TI - Collision and violation involvement of drivers who use cellular telephones. AB - The study sample consisted of 3,869 drivers, split approximately 50/50 between observed cell phone users and those observed not using cell phones (labeled "nonusers"). Cell phone use was determined by a snapshot observation made on city streets. The sample represented 54% of those originally observed, for whom a match was obtained for both vehicle license plate and for gender and estimated age group of the observed driver and that of the driver named in the vehicle policy. Data were obtained from records of insurance claims, police-reported collisions and violations, following a strict protocol to protect individual privacy. The dependent measures were at-fault crash claims and "inattention" violations. A logistic regression model controlled for age, gender, exposure (represented by not-at-fault crash claims), alcohol-related offenses, and aggressive driving offenses. The study also involved a comparison of the contributing factors and collision configurations of police-reported collisions involving the users and "nonusers" in the sample. Drivers observed using cell phones had a higher risk of an at-fault crash than did the "nonusers," although the difference was not significant for males. There was no apparent effect on "inattention" violations. The cell phone users also had a higher proportion of rear-end collisions. The violation pattern of cell phone users suggests that they are, in general, riskier drivers. These differences likely reflect lifestyle, attitude and personality factors. It is essential to control for these factors in assessing the direct risk attributable to cellular telephone use. PMID- 14522662 TI - Modeling accident occurrence at signalized tee intersections with special emphasis on excess zeros. AB - In implementing effective remedial treatments at hazardous intersections, it often is necessary to identify the geometric and traffic factors that lead to accident occurrence. However, one particular problem frequently encountered in accident studies is how to distinguish virtually safe intersections with little likelihood of accident occurrence from those that have happened to have no accident due to the random process. To deal with this problem, the "excess" records of zero accident, the zero-inflated negative binomial was used to assign the probability to the accident outcome. Accident data at 104 signalized tee intersections in Singapore over a period of 9 years were employed for model development. The model indicates that uncontrolled left-turn slip road, permissive right-turn phase, existence of a horizontal curve, short sight distances, large number of signal phases, total approach volume, and left-turn volume may increase accident occurrence. On the other hand, right-turn channelization, acceleration section on the left-turn lane, median railings, and more than 5% approach gradient may reduce accident occurrence. Moreover, there is a trend of reducing accidents over the years. PMID- 14522663 TI - Risk of injury to restrained children from passenger air bags. AB - The objectives of this study were to estimate the prevalence of children's exposure to passenger air bag (PAB) deployments and to determine the relative risk of both minor and more serious nonfatal injuries to restrained children exposed to PABs in frontal impact collisions. Data were collected from 1 December 1998 to 30 November 2001 from a large-scale, child-specific crash surveillance system based on insurance claims, a telephone survey, and on-site crash investigations. Vehicles qualifying for inclusion were State Farm-insured, model year 1990 or newer, and involved in a crash with at least one child occupant < or =15 years of age. Qualifying crashes were limited to those that occurred in 15 states and the District of Columbia. A stratified cluster sample was designed in order to select vehicles (the unit of sampling) for the conduction of a telephone survey with the driver. For cases in which child occupants were seriously injured or killed, in-depth crash investigations were performed. The prevalence of exposure to PABs was calculated as the number of children occupying the right front seat in a PAB deployment crash among all children occupying the right front seat in vehicles equipped with PABs. Complete interview data were obtained on 9,779 vehicles involving 15,341 children. Among PAB-exposed children, 175 (14%) suffered serious injuries versus 41 (7.5%) of those in the comparison group (OR 2.0; 95% CI, 1.1-3.7). The overall risk of any injury (both minor and serious) was 86% among children exposed to PABs, compared to 55% among the comparison group (OR 5.3; 95% CI, 2.1-13.4). Exposure to PABs increased the risk of both minor injuries, including facial and chest abrasions, and more serious injuries, particularly upper extremity fractures. PMID- 14522664 TI - Role of gender in road accidents. AB - Influence of driver sex on road accidents is assessed in this article. Accident records for 3 years and for three different income regions were analyzed. Annual distance traveled, social and economic participation, and effect of public vehicle accidents were considered. Effects of environmental factors and driver age were also included. Driver faults analysis identified possible reasons for accident differences. Analysis of accident severity was used to assess degree of harm. Statistical analysis at the 5% significance level was used to evaluate all differences. The results show that male accident rates are significantly higher. This trend is consistent through all the analyses. Accident differences are significant only in normal driving conditions. Drivers over age 50 had the lowest accident rates. Accident rate differences were caused by lack of attention and impatience among male drivers. Appropriate means of communication should alert concerned populations to these findings. PMID- 14522666 TI - Student health and abuse: what is going on out there? PMID- 14522665 TI - Pedestrian head impact conditions depending on the vehicle front shape and its construction--full model simulation. AB - For the evaluation of pedestrian protection, the European Enhanced Vehicle-Safety Committee Working Group 17 report is now commonly used. In the evaluation of head injuries, the report takes into account only the hood area of the vehicle. But recent pedestrian accident data has shown the injury source for head injury changing to the windshield and A-pillar from the hood. The head contact points are considered to fall on a parallel to the front shape of the vehicle along the lateral direction, but the rigidity of the outer side construction is different from the center area. The purpose of this study is to consider the reason for the change in injury source for recent vehicle models. The head contact points and contact conditions, speed and angle, are thought to be influenced not only by the vehicle's geometry, but also its construction (rigidity). In this study, vehicle pedestrian impact simulations were calculated with a finite element model for several hitting positions, including the outer side areas. Full dummy sled tests were conducted to confirm the simulation results. These results show that, for impacts at the outer sides of the vehicle, the head contact points are more rearward than at the vehicle center. In addition, the speed and angle of the head contact were found to be influenced by the pedestrian height. PMID- 14522667 TI - BEME guide no. 3: systematic searching for evidence in medical education--part 2: constructing searches. AB - Searching for evidence to inform best practice in medical education is a complex undertaking. With very few information sources dedicated to medical education itself, one is forced to consult a wide range of often enormous sources-and these are dedicated to either medicine or education, making a medical education search all the more challenging. This guide provides a comprehensive overview of relevant information sources and methods (including bibliographic databases, grey literature, hand searching and the Internet) and describes when they should be consulted. The process of constructing a search is explained:identifying and combining core concepts, using Boolean algebra and search syntax, limiting results sets, and making best use of databases' controlled vocabularies. This process is illustrated with images from search screens and is followed by numerous examples designed to reinforce skills and concepts covered. The guide has been developed from the ongoing experience gained from the systematic searches conducted for the Best Evidence Medical Education Collaboration, and concludes by looking ahead to initiatives that will shape future searching for medical education evidence. PMID- 14522668 TI - Twelve tips for developing professional attitudes in training. AB - This article is based on a workshop run at AMEE Lisbon, building on work from previous conferences and reported in Medical Teacher (Howe, 2002a). The 30 workshop participants were particularly asked to address the question 'What would you consider essential to include in a medical education curriculum that wishes to teach and assess professional development?'. This question was posed without further constraints, ie. regardless of whether undergraduate or postgraduate, the country or situation of the participant, and the type of setting in which they worked. Participants were invited to consider all aspects of the question, and no assumption was made about the need to reach a consensus. The workshop divided into two groups and shared ideas. This paper presents the main emergent points from discussion, for interest and further collaboration; the level of agreement was considerable, consistent with the peer reviewed literature (Howe, 2002b). The conclusions are therefore shared in the 'Twelve Tips' format as a pragmatic framework for those wishing to review their own curriculum with reference to professional development (PD) issues, or when setting up new opportunities. PMID- 14522669 TI - Developing the teaching instinct. 9: How to teach in an ambulatory care (outpatient) teaching centre. PMID- 14522670 TI - Health and lifestyle needs assessment of medical students in the United Arab Emirates. AB - Medical students represent a significant community investment and promoting their health preserves this investment. In order to design health promotion programs for medical students in the United Arab Emirates, students' needs were assessed by means of a cross-sectional self-administered questionnaire survey. Major findings of the survey included: 14% of students were underweight while 24% of students were overweight or obese; the majority believed their activity levels were insufficient (77%), their stress levels too high (65%) and their diet unhealthy (50%); 33%were not sufficiently active to meet minimum recommended levels; few students (22%) had seen a doctor in the past year and many were unaware of important personal health parameters. When presented with hypothetical health situations, many made inappropriate choices. This survey identified significant health-promotion opportunities for these students that can be carried out during medical school in order to establish a healthier physician population. PMID- 14522672 TI - Stress among medical students in a Thai medical school. AB - This study aimed to determine the prevalence and sources of stress among Thai medical students. The questionnaires,which consisted of the Thai Stress Test (TST) and questions asking about sources of stress, were sent to all medical students in the Faculty of Medicine, Ramathibodi Hospital, Thailand. A total of 686 students participated. The results showed that about 61.4% of students had some degree of stress. Seventeen students (2.4%) reported a high level of stress. The prevalence of stress is highest among third-year medical students. Academic problems were found to be a major cause of stress among all students. The most prevalent source of academic stress was the test/exam. Other sources of stress in medical school and their relationships are also discussed. The findings can help medical teachers understand more about stress among their students and guide the way to improvement in an academic context, which is important for student achievement. PMID- 14522671 TI - A report on student abuse during medical training. AB - The purpose of this study was to determine the incidence, and the consequences, of abusive situations as perceived by students during the course of their medical training. A descriptive study was carried out surveying the entire 2000 fifth year class of 181 in the Medical School of the University of Chile. The questionnaire was answered by 144 students. Results showed that 91.7% of the students who responded had suffered at least one episode of abuse while enrolled in medical school. The main offenders were teachers and peers. Verbal abuse was the most common (85.4%), followed by psychological (79.9%), sexual(26.4%) and physical (23.6%) abuse. Students reported that abuse had effects on their mental health, social life and the image they had of physicians; 17% considered dropping out of school as a consequence of this experience. Efforts should be addressed to prompt educators to reflect on their role. PMID- 14522673 TI - Standards in medical education in the European Union. AB - Standards in medical education are necessary to enable evaluation of programmes and comparison between institutions. EC Directive 93/16 lays down the standards for both the duration and content of basic medical education in the European Union but the directive is open to wide variations in interpretation. A new approach is needed and the development of common learning outcomes is suggested as a possible solution. PMID- 14522674 TI - Web-based evaluation of medical clerkships: a new approach to immediacy and efficacy of feedback and assessment. AB - The classical assessment of medical students has been based on the 'apprenticeship' model, which consists of written examinations and tutor-led assessment of coursework; this method has proved to be very dependent on tutors' attitudes and instructional skills and on patients' availability. This paradigm has shifted in recent years as a result of the new implementation of modular assessments, logbooks and portfolios during clerkship rotations. Portfolios have been demonstrated to bea very useful tool to promote self-reflection, prompt feedback and skills development. However, written portfolios not only introduce additional paperwork for both students and tutors but also have some limitations for immediacy and effectiveness of feedback. To obviate these limitations an electronic portfolio is proposed here to be used by students and tutors during clinical clerkships. Based on the principles for good practice in undergraduate education the authors show the advantages of this method and its multiple applications to promote students' development of skills and attitudes and to improve tutors' acceptance of this innovative evaluation method. PMID- 14522675 TI - What do graduates think about a two-week rational pharmacotherapy course in the fifth year of medical education? AB - The present study aims to assess the short- and mid-term post-graduation impact of a pharmacotherapy course in the fifth year at Marmara University School of Medicine by an objective (OSCE) and a subjective (questionnaires) evaluation. Statistical comparison of pretest, posttest-exposed case and posttest-unexposed case scores indicated both a retention and a transfer effect of training. The post-course questionnaire revealed that 95%of the students found the course useful and necessary; 97% reported that they will apply a rational pharmacotherapy approach using this model and communicate better with their patients. The post-graduation questionnaire also showed that the majority of them have learned general principles of rational pharmacotherapy(90%), gained good prescribing (90%) and communication skills (87.5%), and understood the importance of non-pharmacological treatment alternatives (100%). In general, they stated that they would apply the principles during their medical practice and they believed their colleagues would do too. In conclusion, the present study demonstrates the benefit of a clinical pharmacology programme focused on rational pharmacotherapy during the clinical years of medical education. PMID- 14522676 TI - Students' perspectives on the educational environment, Faculty of Medical Sciences, Trinidad. AB - The Dundee Ready Education Environment Measure (DREEM) was administered to 70 final-year medical students and 36 first-year medical interns (pre-registration house officers). The overall total mean DREEM scores for the five subscales namely, students' perceptions of the atmosphere, students' perceptions of learning, students' social self-perceptions, students'perceptions of teachers and students' academic self-perceptions-was 109.9 and the total mean scores for the subgroups-male students, male interns, female students and female interns-were 103.39, 111.82, 111.33 and 113.15, respectively. The lowest scores were assigned to students' social self-perceptions and students' perceptions of the atmosphere. All of the participants except the male interns recorded the highest scores for the subscale academic self-perceptions. PMID- 14522677 TI - The development and evaluation of a fieldwork educator's training programme for allied health professionals. AB - This paper describes the development, implementation and evaluation of a multi professional fieldwork educator's course. The study explored the current provision of such training available to a range of healthcare professionals within the Greater Manchester West Consortium boundaries and the extent to which the core skills needed could be provided within a multi-professional programme. The methods used included surveys, focus groups, rating scales completed by participants immediately following the pilot course, a focus group of participants after three months of the course and a nominal group six months after completion. This study demonstrates that the skills needed by a fieldwork educator can be taught in a multi-professional programme, allowing the practitioner to assimilate her/his skills into her/his own working environment, and outlines issues to be taken into consideration when designing such courses. PMID- 14522678 TI - One-on-one tutorials in private practices and clinics: four years of experience in Basel, Switzerland. AB - The one-on-one tutorial is a new form of learning that is practice oriented. It is based on a teacher-student relationship continuing over two years. Since 1997, third- and fourth-year students have worked for one half day per week under the supervision of their tutor, be it in a private practice or in a hospital. This programme facilitates direct patient contact at an early stage of medical school. In addition, it allows students to apply their knowledge in everyday life. The interactive form of learning is of paramount importance in this module. The ARIVA learning model was developed specially for third-year students and the logbook for fourth-year students. After each tutorial third-year students completed the ARI VA worksheet and fourth-year students completed the logbook. They were handed in together with the structured learning report. Between 85 and 109 students per year participated in these tutorials, totalling 733 students. Each student was taught an average of 3.1 patients in the presence of the tutor. In addition students examined an average of 2.4 patients independently and fulfilled an average of 1.2 practical tasks. For the fourth-year students the number of contacts with patients and the spectrum of diseases examined are impressive. All learning goals were fulfilled The one-on-one tutorial is a practice-oriented, interactive learning method. It uses a variety of didactic methods based on the principals of problem-oriented learning. In a relatively early stage of their medical education one-on-one tutorials give students the opportunity to learn independently how to interview and examine patients. They also give students the possibility to acquaint themselves with a multitude of diseases with the aid of instructions and demonstrations. They are conducive to work with patients above all because students learn how to perform certain techniques. PMID- 14522679 TI - Consultation by the United Kingdom to South Africa to develop human diversity teaching. AB - This paper describes consultation provided by the University of Leicester to the Medical University of Southern Africa (Medunsa) funded through British Council and the Department for International Development to develop a course addressing issues of diversity at Medunsa. The aims of the link were essentially for Leicester to provide support and guidance on the development of aspecific module. This inevitably included the provision of training in key areas such as teaching skills and evaluation of teaching. This paper focuses on the potential difficulties of such a link, which included cultural differences, issues of hierarchy and student influence. The project outcomes have been the development of a coherent module on human diversity which is assessed in such a way as to support the development of student writing skills, increased staff teaching skills and a joint conference to disseminate the learning that has taken place from the project. PMID- 14522680 TI - Teaching problem solving and decision making in undergraduate medical education: an instructional strategy. AB - Actual changes in the context of real care along with their impact on current practice modes prompted the authors to develop a new instructional strategy, which aims to improve the diagnostic reasoning competence of medical undergraduates. In this strategy the use of visual representations is promoted. A diagnostic panorama represents differential diagnosis. A diagnostic diagram exposes the diagnostic reasoning process. This paper outlines the strategy and its assumptions. PMID- 14522681 TI - Utilizing the SPICES model to evaluate the continuous improvement of a clinical development programme facilitated by Clinical Squadron, 22 Field Hospital. PMID- 14522687 TI - Potential of commonly consumed green leafy vegetables for their antioxidant capacity and its linkage with the micronutrient profile. AB - Green leafy vegetables (GLV) offer a cheap but rich source of a number of micronutrients and other phytochemicals having antioxidant properties. The potential of 30 GLV in the raw and cooked form as natural antioxidant supplements for vegetarian diets was assessed. There was a large variability in the values of antioxidant activity of various GLV extracts in the lipid micelles (1.5-5.6 mM vitamin E/100 g for raw samples and 1.6-3.8 mM vitamin E/100 g for cooked samples). Similar to thiobarbituric acid reactive substances values, the super oxide scavenging ability values also exibited large variation (10.6-55.9), with significantly higher values in the raw state than the cooked state (P<0.001). Omum leaves, radish leaves and lettuce had high values for this index. The range of values for ferrous iron chelating activity was from 9.3 to 65.7 mM EDTA/100 g food material, indicating again a large variability in this assay. Leaves of coriander, amaranthus viridis, colcasia green and drumstick showed high values, while Amaranthus p. Colocasia black and amaranthus red exibited low values. Differences between raw and cooked values were highly significant for all the three indices (P<0.001). PMID- 14522686 TI - Mulberry (Morus alba) leaves as human food: a new dimension of sericulture. AB - Mulberry leaf is commonly used for sericulture in almost every part of the world but its potential to be utilized for human consumption is not well recognized. This paper deals with development of mulberry leaf powder and its use with wheat flour to develop paratha, the most common food item of breakfast and dinner in the Indian diet. The optimum ratio of the mulberry leaf powder and wheat flour (MLP-WF) mix for preparation of paratha on the basis of sensory quality was found to be 1:4. The protein quality of the MLP-WF mix was estimated by measuring the Protein Efficiency Ratio, and was found to be 1.82 against a casein diet for which a value of 2.44 was observed. The in vivo toxic effect of mix was studied and no adverse effect on the growth of internal organs of rats (heart, liver, kidney and testes) was found. The storage stability of the mix was estimated for a period of 2 months in polyethylene bags at room temperature. A non-significant difference was observed between paratha prepared from fresh and stored mix. This indicated that mix can be stored for a period of 2 months at room temperature without loss of quality. PMID- 14522688 TI - Inter-relationship between electrophoretic characteristics of pseudocereal and cereal proteins and their microscopic structure for possible substitution based on nutritional evaluation. AB - Amaranth, soybean and maize were screened for proteins and their nutritional value. Isopropanol-soluble protein and buffer-soluble protein fractions were extracted from seeds and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The comparison of the identity and differences between investigated plants was carried out by the obtained SDS-PAGE electrophoretic patterns, and their microstructure was determined by scanning electron microscopy. Electrophoretic patterns of extracted proteins have shown that the main protein subunits were concentrated between 10 and 50 kDa. Variations were found in major fractions and minor bands as well as in the fine structure. The microstructure of pseudocereal and cereal protein fractions was inter-related with the results obtained by their electrophoretic separation. Pseudocereal amaranth can be used as a nutritive substitute of cereal maize in functional foods. PMID- 14522689 TI - Effect of addition of ripe bananas on some physico-chemical properties of maize 'extract'. AB - Flour mixes obtained by the addition of banana pulp in various proportions (0 50%) to maize 'extracts' were evaluated for some quality characteristics. All the mixes had significantly lower values of crude protein, fat and water-holding capacity. Gelation, however, significantly increased the water-holding capacity in all cases. The ash content, titratable acidity and total sugars increased tremendously with an increase in the level of banana substitution. While both Adam's consistency values and equilibrium moisture content decreased with an increase in the level of banana substitution, the syneresis values did not show any consistent pattern. The consistently low moisture content and the results of the moisture sorption isotherms suggest a good storage stability of all the mixes, especially when kept under conditions of water activity of 0.30 and below, and their possible suitability for baked products. PMID- 14522690 TI - Traditional non-alcoholic beverage, Togwa, in East Africa, produced from maize flour and germinated finger millet. AB - The traditional non-alcoholic beverage in East Africa, togwa, produced from the flour of maize and germinated finger millet (finger millet malt), was investigated. The preparation techniques of togwa observed in the rural villages of East Africa are described, and the temperature and pH profile of togwa during its manufacture are also shown. Maize and finger millet malt should be the source of starch and amylase, respectively. Maize flour slurry was heated once with stirring up to around 80 degrees C and cooled to about 50 degrees C, and then finger millet malt flour was added to the warm porridge paste and kept at about 50 degrees C for 20 min. The consistency of the paste was suddenly reduced by addition of finger millet malt flour and the gel paste changed to viscous liquid, which was kept in a container and incubated at ambient temperature for 15 h. After incubation it became sweet and was ready to drink without removal of any insoluble materials. Changes in the concentration of glucose and lactic acid of togwa during its maturation period were measured using portable devices until 70 h incubation at the rural village of Tanzania in a dry season. Glucose level increased with incubation and reached the threshold value of sweetness; 24 h incubation later, the lactate level increased and pH decreased. The preparation techniques of a traditional alcoholic beverage, pombe, were also investigated in the same rural locality, and the differences and characteristics of both traditional beverages are discussed. PMID- 14522691 TI - Estimated nutrient intakes and adequacies in Bangladesh change when newer values for vitamin A, iron and calcium in commonly consumed foods are applied. AB - A food consumption survey was conducted in rural Bangladesh in January-March 1996 using a 24-h food weighing method. Intakes and adequacies of energy, vitamin A, vitamin C, iron and calcium were estimated for 81 children (aged 24-71 months) and 182 women (aged 18-45 years) using three sets of food composition tables. The local food composition table was modified with newer values for nutrient contents of foods identified as important for the contribution of vitamin A and iron. Significant differences in estimated dietary intakes and adequacies were found for vitamin A and iron when different sets of food composition values were used. This study shows that up-to-date chemical analyses of foods selected on the basis of a high frequency of consumption and a large contribution of the nutrients in focus is a useful approach in improving the quality of food composition tables, resulting in greater accuracy of estimates of nutrient intakes and adequacies. PMID- 14522692 TI - Sexual dimorphism in fat patterning in a sample of 5 to 7-year-old children in Oxford. AB - It is now well recognised that, in addition to total body fat, fat distribution is a major risk factor for cardiovascular disease and insulin resistance/diabetes in both adults and children. Traditionally, sexual dimorphism in fat patterning has been regarded as occurring at puberty. The aim of this study was to investigate gender differences in fat patterning in a group of children in Oxford. Anthropometric data was collected for 95 girls and 93 boys aged 5-7 years. Body mass index, percentage body fat and fat patterning indices were calculated using skinfold thickness measurements. While girls at this age had significantly larger percentage body fat and skinfolds, the fat patterning indices showed no differences between genders and no indication of greater truncal adiposity. The boys had a larger mean waist-hip ratio (0.96+/-0.04) compared with the girls (0.93+/-0.04) (P<0.001). The larger ratio in boys was the outcome of the smaller hip circumference in the boys reflecting less gluteal adiposity. This suggests that even at this young age there is clear evidence of sexual dimorphism in fat patterning, with girls showing greater subcutaneous adiposity mainly contributed by gluteal fat. The present study confirms the findings of American and European workers that sexual dimorphism of fat patterning in children is present at 5-7 years of age. Second, that these differences in fat patterning can be detected using simple anthropometric measurements such as hip circumference and skinfold thicknesses. The significance and importance of hip circumference measurements in predicting health outcome is an area that merits further investigation. PMID- 14522693 TI - Nutritive value of meals, dietary habits and nutritive status in Croatian university students according to gender. AB - The aim of this study was to evaluate daily menus at students' restaurants and to report dietary habits and other health-related behaviour of Croatian university students (n=2075) according to gender. A specially designed self-administered questionnaire was used. One hundred and twenty daily menus were chosen by random sampling, and the nutritive value was calculated using food composition tables. Daily menus on average provide an adequate amount of energy, protein and most micronutrients: 88.2% of daily menus provide a balanced intake of protein, fat and carbohydrates, 22.5% of daily menus provide more than 300 mg of cholesterol, and 58.8% have more than 25 g dietary fibre. On average, students had 2.4 meals and 1.3 snacks per day. Breakfast was the most often skipped meal. Red meat, cereals and fast food were consumed more often by males (P<0.05). Low-fat dairy products, whole grain products and breakfast cereals were consumed more often by females (P<0.05). The most common choice for snacks was fruit. Males exercised more than females (4.4 h/week versus 1.6 h/week; P<0.05). A higher percentage of females (29.8%) than males (17.2%) smoked cigarettes. For alcohol consumption it was vice versa: 88.9 and 84.8% of males and females, respectively. A total of 80.4% of students were well nourished. This study showed that meals offered at students' restaurants are adequate. Dietary and other health-related behaviour differed according to gender. Clustering of some behaviours was observed. PMID- 14522694 TI - Race-, gender- and age-specific differences in dietary micronutrient intakes of US children. AB - Race-, gender- and age-specific differences in dietary micronutrient intakes of 1 to 10-year-old US children were evaluated. Three-day, dietary intakes from the US Department of Agriculture's Continuing Survey of Food Intakes by Individuals were evaluated. Data from 1895 children (967 males, 928 females; 1,540 Whites, 355 Blacks) who resided in the 48 conterminous states were analyzed. Micronutrient intakes, intakes as percent of the Recommended Dietary Allowance (RDA) and percent of children who consumed < or =67% of the RDA were computed. Black males compared with White males, Black females compared with White females and White females compared with White males had significantly lower dietary intakes for several micronutrients. More Black males than White males had intakes < or =67% of the RDA for vitamin E, calcium and zinc. Blacks and female children were at a greater risk for vitamin A, vitamin E, calcium, iron and zinc deficiency. PMID- 14522695 TI - Demographic and lifestyle factors associated with body mass index among children and adolescents. AB - Cases of overweight and obesity among adults, adolescents, and children are escalating in the United States. Using data from the Continuing Survey of Food Intake for Individuals (CSFII) 1994-1996 and 1998, and the National Health and Nutrition Examination Survey III (NHANES), we developed multivariate regression models to examine the association between demographic and lifestyle variables (gender, race/ethnicity, age, family income, diet, and television viewing) and the body mass index (BMI) of children aged 6-11 years, and adolescents aged 12-19 years for CSFII and aged 12-16 years for NHANES. In the children's model, the statistically significant demographic predictors of BMI were age, race, gender, and family income. Among adolescents, the demographic predictors were age, race, and gender. Among the lifestyle variables, dietary factors were not associated with BMI in children. On the contrary, carbohydrates (less added sugars) had a statistically significant inverse relationship to BMI among adolescents. In both children and adolescents, television viewing was markedly associated with BMI, and participation in team sports was negatively associated with BMI. The overall process by which overweight develops is complex, and our four models explained only a small portion of total variance of BMI. The CSFII models explained 8.5% of the variance for children and 11.4% of the variance for adolescents. The NHANES models explained 8.8% of the variance for children and 12.5% of the variance for adolescents. Demographic variables accounted for roughly one-half of the explained variance for both children and adolescents. The rest of the explained variance was accounted for by television viewing, diet, and family income. PMID- 14522696 TI - Skipping breakfast is associated with dysmenorrhea in young women in Japan. AB - To evaluate the effects of breakfast habits on the quality of life in young women in Japan, the relation between breakfast habits and menstrual and intestinal disorders was estimated by responses to a questionnaire. Subjects were recruited from young Japanese female students of Ashiya College ranging in age from 18 to 20 years. Breakfast habits were classified into three groups (group I, having breakfast every morning, n=289; group II, having breakfast one to six times per week, n=106; group III, having breakfast less than once a week, n=44). The intensity of dysmenorrhea was classified into three grades (score 1, free of pain or painful, but without need for analgesic; score 2, painful, requiring analgesic; score 3, painful, not relieved by analgesic). The grades of bowel movement were classified into three groups (score 1, no more than once a week; score 2, two to six times a week; score 3, every day). Group II and group III had significantly higher scores for dysmenorrhea compared with group I. Students in group III showed a tendency toward constipation. Since dysmenorrhea is a risk factor for psychological disorders as well as gynecological diseases, the present study suggests the importance of eating breakfast to the quality of life of young women. PMID- 14522701 TI - Investigations of Enterococcus faecalis-induced bacteraemia in brown layer pullets through different inoculation routes in relation to the production of arthritis. AB - In the present aerosol experiment, assessment of the respiratory tract of 1-day old birds as a natural route of infection for induction of Enterococcus faecalis bacteraemia and arthritis was performed. Second, the severity and type of arthritis produced through intramuscular infection in two different inoculation sites (musculus pectoralis versus musculus gastrocnemius) was studied. Third, the resulting bacteraemia was assessed qualitatively and quantitatively in relation to the occurrence of arthritis. Exposure of 1-day-old brown layer pullets to aerosolized E. faecalis with an estimated uptake of 10(4) to 10(5) colony forming units per chick resulted in bacteraemia; however, joint lesions were not induced. In contrast, 3/10 birds inoculated intratracheally with 10(8) colony forming units developed both bacteraemia and arthritis. This suggests the occurrence of a dose effect and a role for the respiratory tract as a natural infection route in young chickens. In both intramuscularly inoculated groups the incidence of arthritis was 10/10 birds and 9/10 birds, respectively. Birds inoculated in the m. pectoralis developed symmetric polyarthritis, which harmonizes with haematogenous colonization of joints. In contrast, m. gastrocnemius-inoculated chicks mostly had asymmetric (poly)arthritis of the injected leg and varus deformation of the contralateral leg, suggesting predominantly local spread. The qualitative and quantitative results of bacteriology of blood samples show that arthritis develops in those groups with the highest number of bacteraemic birds and the highest median bacterial colony forming units per millilitre of blood during the first 24 to 36 h after treatment. PMID- 14522700 TI - Avian host range of Chlamydophila spp. based on isolation, antigen detection and serology. AB - Published reports and our own diagnostic data on the avian host range of avian Chlamydophila spp. are presented in an attempt to provide evidence for the large number of bird species that have been naturally infected with chlamydia. The term 'chlamydia-positive' is based on either isolation of the organism and antigen detection or on serological detection of circulating antibodies. The list of chlamydia-positive birds contains the six major domestic species (chicken, turkey, Pekin duck, Muscovy duck, goose, and pigeon), the three minor domestic species (Japanese quail, bobwhite quail, and peafowl) and a total of 460 free living or pet bird species in 30 orders. The order Psittaciformes contains by far the most (153 of 342; 45%) chlamydia-positive bird species. More than 20% of all species per order are positive for chlamydia in the orders Lariformes (gulls, 26 of 92 species; 28%), Alciformes (alks, six of 23 species; 26%), Sphenisciformes (penguins, four of 16 species; 25%), and Anseriformes (ducks and geese, 33 of 157 species; 21%). Only 5% of all bird species (14 of 259 species) in the order Phasianiformes (gallinaceus birds) are chlamydia-positive. The different percentages of chlamydia-positive bird species reflect: (i) a high rate of investigations (e.g. of domestic birds) compared with infrequent testing (e.g. of Charadriiformes or Cuculiformes), (ii) frequent zoonotic implications (e.g. psittacine and columbiform birds), and (iii) an assumed high susceptibility to infection and subsequent seroconversion (e.g. waterfowl). PMID- 14522702 TI - Ability of Massachusetts-type infectious bronchitis virus to increase colibacillosis susceptibility in commercial broilers: a comparison between vaccine and virulent field virus. AB - The abilities of Massachusetts-type vaccine virus and virulent infectious bronchitis (IB) field virus to increase colibacillosis susceptibility were compared. In four experiments, 29-day-old female commercial broilers housed in isolators, were infected intratracheally and oculonasally with IB vaccine strains (H120 and H52) or virulent IB field strains (D387 and M41) (4.8 or 6.8 log(10) median embryo infective dose, per broiler). Five days later, Escherichia coli 506 strain was given intratracheally (5.6 to 8.8 log(10) colony forming units/broiler). The incidence of nasal discharge at 3 and 5 days after IB virus infection was used to assess the clinical effect of the IB infection, while mortality, body weight uniformity and E. coli lesions at 7 days following E. coli inoculation were used as parameters for colibacillosis. Nasal discharge was observed in 6/117 (5%), 26/119 (22%), 35/119 (29%) and 115/120 (96%) of broilers infected with H120, H52, D387 and M41 virus, respectively. Apart from H52 and D387, differences between IBV strains were significant. IB vaccine and virulent IB viruses did not generally differ significantly in their ability to induce colibacillosis susceptibility. Mean colibacillosis lesion scores of H52-infected birds even significantly exceeded those of birds infected with the other IB viruses. The ability of H120 virus to induce colibacillosis susceptibility tended to be the weakest. The practical consequences of these findings are discussed. PMID- 14522703 TI - Association between in vitro heterophil function and the feathering gene in commercial broiler chickens. AB - We recently showed that in vitro heterophil functional efficiency in commercial broiler chickens is genetically controlled and may be a sex-associated trait. To further characterize the genetic mechanism(s) of heterophil functional efficiency, we wanted to determine whether the feathering gene, present on the Z sex chromosome, contributes to heterophil functional efficiency. Heterophils from two pairs of broiler lines were evaluated; each pair contained a fast feather (FF) (lines A and X) and a slow feather (SF) line (lines B and Y). On days 1 and 4 post-hatch, heterophils isolated from two sets of pure line broilers (A and B, and X and Y) were evaluated for their ability to (1) phagocytize Salmonella enteritidis, and (2) exhibit bactericidal activity against S. enteritidis. On days 1 and 4 post-hatch, heterophils isolated from the FF lines were statistically (P < or = 0.02) more proficient at phagocytizing S. enteritidis than heterophils from SF lines. Bactericidal activity was also statistically (p < or = 0.02) greater on day 1 post-hatch in the heterophils isolated from FF lines compared to heterophils isolated from SF lines. These data indicate that the presence of the FF gene locus on the Z sex chromosome contributes to heterophil function and may contribute to the early innate immune competence of a flock. PMID- 14522704 TI - Direct (non-vector) transmission of West Nile virus in geese. AB - During a recent epizootic, losses due to West Nile virus (WNV) infection in young goose flocks were estimated to be far greater than expected if mosquito-borne transmission was the principal route of infection. Contact transmission was investigated experimentally as an alternative explanation. A group of 10, 3-week old geese were inoculated subcutaneously and placed in one insect-proof room with 20 geese of the same age. A group of 10 geese were housed in an adjacent insect proof room to serve as an environmental control. All geese in the inoculated group produced antibodies, eight became viraemic and five died between 7 and 10 days after infection. Virus was shed from the cloaca and oral cavity by three geese. Two of the in-contact birds died on days 10 and 17 after infection, and WNV was recovered from another three birds. None of the environmental control group became infected. This result strongly suggests that horizontal transmission of WNV can occur in commercial flocks and may be aggravated if cannibalism and feather-picking of sick geese occur. PMID- 14522705 TI - Effects of cyclosporin A on the immune responses and pathogenesis of a virulent strain of Mycoplasma gallisepticum in chickens. AB - Immune responses to the virulent S6 strain of Mycoplasma gallisepticum in immunocompetent and cyclosporin A (CsA)-treated specific pathogen free chickens were investigated, and pathogenesis of the M. gallisepticum strain was also examined. Ten-day-old specific pathogen free chickens were inoculated by eye-drop with M. gallisepticum, and a control uninfected group was inoculated with mycoplasma broth. Blood was collected weekly for 4 weeks from five birds in each group and whole blood lymphocyte transformation assayed against concanavalin A and lipopolysaccharide. Blood samples were also collected at intervals for serological assays. Live body weight, clinical signs and lesions were monitored, and recovery of M. gallisepticum was attempted from choanal cleft of live birds and also from various sites at necropsy. In parallel to the aforementioned groups, another set of two groups of chicks treated with CsA was infected with M. gallisepticum S6 or mycoplasma broth. These groups were subjected to the same experimental procedures. In the immunocompetent chickens, M. gallisepticum caused temporary T-cell suppression at 2 weeks post-infection. Comparison of the clinical signs and macroscopic lesions produced in immunocompetent and CsA treated chickens indicated that T cells may not play an active role in disease development. The percentage of birds with mycoplasma isolation and the load of mycoplasmas suggested that T cells may have some role in resisting mycoplasma colonization or in the elimination of the infection. PMID- 14522706 TI - Prevalence and transmission of haemolytic Gallibacterium species in chicken production systems with different biosecurity levels. AB - A stratified cross-sectional study consisting of four strata of biosecurity based on production system type, including organic/free-range layer, battery-cage layer, layer parent, broiler parent and broiler grandparent flocks, was performed to estimate the prevalence of haemolytic Gallibacterium spp. Thirty birds were sampled by tracheal and cloacal swabs in each flock. A flock was considered infected when just one bird tested positive. A total of 27 flocks was included in the study. All chickens from the broiler grandparent flocks sampled negative, whereas 28% of the broiler parents, 40% of the layer parents, 67% of the battery cage layers and 96% of the organic/free-range chickens sampled positive. A total of 95.9% (standard deviation +/- 7.6%) of birds from infected flocks was colonized by haemolytic Gallibacterium species. A significantly higher number of tracheal swabs was positive compared with cloacal swabs. The probability of vertical transfer was investigated by sampling offspring from an infected as well as a non-infected parent flock. None of the samples were found positive. In conclusion, we showed that haemolytic Gallibacterium spp. were widely distributed within the Danish commercial chicken production systems. However, prevalence proportions were highly influenced by the production system and found to be significantly associated with the biosecurity level observed in the flocks. In general, flock infections resembled an 'all or none' type of colonization as practically all of the chickens in infected flocks sampled positive. There was no evidence of vertical transmission of Gallibacterium. PMID- 14522707 TI - Characterization of infectious bursal disease virus isolates from Indonesia indicates the existence of very virulent strains with unique genetic changes. AB - Sequencing of the hypervariable region of viral protein VP2 of infectious bursal disease virus (IBDV) isolates obtained from non-vaccinated chickens in Indonesia showed that the majority (16/17) were closely related to published very virulent (vv)IBDV strains. Four isolates contained identical amino acid sequences to Asian and European vvIBDVs, sharing vv-specific amino acid residues 222(Ala), 256(Ile), and 294(Ile). Eight isolates differed by one amino acid at position 222(Ala- >Ser); however, this change did not alter the pathogenicity or antigenicity of these strains. Two isolates, with amino acid substitutions at positions 272(Ile- >Thr) and 279(Asp-->Asn), did not cause clinical disease or mortality, and were therefore considered to be naturally occurring, attenuated mutants of vvIBDV. The results illustrate variability that might occur among vvIBDV strains. PMID- 14522708 TI - Evaluation of a nucleoprotein-based enzyme-linked immunosorbent assay for the detection of antibodies against infectious bronchitis virus. AB - As an immunogen of the coronavirus, the nucleoprotein (N) is a potential antigen for the serological monitoring of infectious bronchitis virus (IBV). In this report, recombinant N protein from the Beaudette strain of IBV was produced and purified from Escherichia coli as well as Sf9 (insect) cells, and used for the coating of enzyme-linked immunosorbent assay (ELISA) plates. The N protein produced in Sf9 cells was phosphorylated whereas N protein from E. coli was not. Our data indicated that N protein purified from E. coli was more sensitive to anti-IBV serum than the protein from Sf9 cells. The recombinant N protein did not react with the antisera to other avian pathogens, implying that it was specific in the recognition of IBV antibodies. In addition, the data from the detection of field samples and IBV strains indicated that using the recombinant protein as coating antigen could achieve an equivalent performance to an ELISA kit based on infected material extracts as a source of antigen(s). ELISAs based on recombinant proteins are safe (no live virus), clean (only virus antigens are present), specific (single proteins can be used) and rapid (to respond to new viral strains and strains that cannot necessarily be easily cultured). PMID- 14522710 TI - Anticoccidial vaccination: the absence or reduction of numbers of endogenous parasites from gross lesions in immune chickens after virulent coccidial challenge. AB - Male and female floor-reared chickens were immunized with a live, attenuated anticoccidial vaccine (Paracox) and then, 28 days after vaccination, were challenged with virulent strains of Eimeria acervulina, Eimeria brunetti, Eimeria maxima, Eimeria necatrix or Eimeria tenella. The relationship between the post challenge growth rate and specific gross lesion grades, matched in individual birds, was examined. The numbers of endogenous parasites associated with graded lesions of each species were compared with those in challenged naive control birds. The vaccinated chickens were clinically immune judged by weight gains, but the naive controls reared under the same conditions were unprotected. The mean weight gains of vaccinated, immune birds had lower coefficients of variation than those of unvaccinated, unprotected birds challenged with the same species. Whichever Eimeria species the birds were challenged with, a proportion (5 to 75%) of the challenged, vaccinated, immune birds exhibited some coccidial lesions, mostly graded 1 or 2 (on a scale of 0 to 4); whereas all unvaccinated, unprotected birds had severe coccidial lesions, mostly graded 4. Innumerable endogenous parasites were associated with all lesions seen after challenging unvaccinated, naive birds, but 68% of the gross lesions in challenged, vaccinated, immune birds had no associated parasites, and the remaining 32% had very few. It is concluded that the use of lesion grades alone to assess an anticoccidial vaccine may under-rate its efficacy, and that the occasional presence of gross lesions in commercially vaccinated chickens does not indicate vaccine failure unless performance is also adversely affected. Although in a naive bird gross lesions usually indicate disease, the presence of any gross lesions in a bird judged by performance criteria to be immune may be interpreted as the host's successful repulsion of a parasite challenge. PMID- 14522709 TI - Diagnosing Clostridium perfringens-associated necrotic enteritis in broiler flocks by an immunoglobulin G anti-alpha-toxin enzyme-linked immunosorbent assay. AB - The tools available for monitoring necrotic enteritis caused by Clostridium perfringens in broiler chickens have been limited, particularly for identifying subclinical disease. In this study, a modified enzyme-linked immunosorbent assay was used to quantify levels of specific immunoglobulin G to C. perfringens alpha toxin in serum from broilers. We found significantly higher antibody levels in broilers with a history of subclinical necrotic enteritis compared with a zinc bacitracin-treated group with a low level of gut lesions. Furthermore, in 4.5 week-old commercial broiler flocks, there was an association between the occurrence of C. perfringens-associated hepatitis at slaughter and the immune response to alpha-toxin. Practical solutions for defining cut-off levels for positive serum samples at individual and flock levels are proposed, and were found to be useful on a set of samples available from flocks with different histories regarding the occurrence of C. perfringens-associated disease. This serological approach seems promising as a diagnostic tool in research and disease monitoring regarding C. perfringens-associated disease. PMID- 14522711 TI - Detection of egg drop syndrome virus antigen or genome by enzyme-linked immunosorbent assay or polymerase chain reaction. AB - Mouse monoclonal antibodies (mAbs) were produced against an Indian isolate of egg drop syndrome (EDS) virus and characterized. Four hybridoma clones were secreting mAbs that bound to a 100 kDa protein, presumably the hexon protein. These mAbs were found to cross-react with two other Indian isolates of EDS virus and to the reference UK 127 strain. Three of these mAbs were mapped to the same epitope compared with the other mAb (F8), which bound to a different epitope. An antigen capture enzyme-linked immunosorbent assay (AC-ELISA) was developed using the F8 mAbs as capture antibody and polyclonal chicken serum against EDS virus as detection antibody. A polymerase chain reaction (PCR) was used to detect the EDS viral genome. Following experimental infection of oestrogen-treated chickens with EDS virus, cloacal swabs, oviduct, uterus and spleen were collected at different days post-infection and used in both AC-ELISA and PCR, directly and after a single passage in embryonated duck eggs. The sensitivity and specificity of antigen detection by AC-ELISA or PCR was 95% and 98%, respectively. For diagnosis of EDS viral infections, PCR is recommended due to its ease and the lack of requirement of prepared reagents such as mAbs or conjugates. We recommend that PCR be performed directly on boiled tissue homogenates. Any negative samples may be passaged in embryonated duck eggs and the allantoic fluids tested by PCR before a conclusive negative diagnosis is given. PMID- 14522712 TI - H9N2 influenza viruses prevalent in poultry in China are phylogenetically distinct from A/quail/Hong Kong/G1/97 presumed to be the donor of the internal protein genes of the H5N1 Hong Kong/97 virus. AB - Ten H9N2 influenza virus strains isolated from diseased chickens in different farms in China during 1995 to 1999 were antigenically and genetically characterized. The haemagglutinins of the isolates were not related to those of A/quail/Hong Kong/G1/97 (H9N2) (Qa/HK/G1/97), but were closely related to that of A/chicken/Hong Kong/G9/97 (H9N2) (Ck/HK/G9/97). The neuraminidase of these isolates had a deletion of three amino acid residues at positions 63 to 65 as compared with those of Ck/HK/G9/97, while that of Qa/HK/G1/97 lacked two amino acids at positions 38 and 39. The PB2 genes of the isolates were not related to those of Qa/HK/G1/97 or Ck/HK/G9/97, but showed some relationship to that of A/duck/Hong Kong/Y439/97 (H9N2) (Dk/HK/Y439/97). The PB1 genes of the isolates were not related to those of the three representative strains. The PA, NP, M, and NS genes of the isolates belonged to the same lineage as those of Ck/HK/G9/97, and were distinct from those of Qa/HK/G1/97 and Dk/HK/Y439/97. The present results indicate that H9N2 influenza viruses prevalent in chicken populations in China belong genetically to one lineage and are distinct from Qa/HK/G1/97, presumed to be the donor of the internal protein genes of the highly pathogenic H5N1 influenza virus in Hong Kong in 1997. PMID- 14522714 TI - Rocky Mountain ecosystems: diversity, complexity and interactions. AB - The interior west of North America provides many opportunities to study ecosystem responses to climate change, biological diversity and management of disturbance regimes. These ecosystem responses are not unique to the Rocky Mountains, but they epitomize similar scientific problems throughout North America. Better management of these ecosystems depends on a thorough understanding of the underlying biology and ecological interactions of the species that occupy the diverse habitats of this region. This review highlights progress in research to understand aspects of this complex ecosystem. PMID- 14522715 TI - Diversity, complexity and interactions: an overview of Rocky Mountain forest ecosystems. AB - This overview of Rocky Mountain forest ecosystems characterizes some of the major types, processes and management issues in the region. There are large ranges in edaphic conditions and striking environmental gradients, all interacting to influence the distribution of species, the nature of communities and disturbance regimes. The discussion focuses on the central role of disturbance and how understanding disturbance regimes influences the search for effective approaches to stewardship. PMID- 14522716 TI - Another perspective on altitudinal limits of alpine timberlines. AB - Recent hypotheses of timberline causation include the possibility that limitations to growth processes may be more limiting than restrictions on photosynthetic carbon gain, and that cold soil is a primary limiting factor at high altitude. However, almost all of the supporting data for timberline causation have come from studies on older trees, with little focus on the mechanisms of seedling establishment and the growth of saplings away from the forest edge into the treeline ecotone. We describe a conceptual model of timberline migration that invokes a strong dependence on ecological facilitation, beginning with seed germination and continuing through seedling establishment and sapling growth to the stage where trees with forest-like stature form new subalpine forest at a higher altitude. In addition to protection from severe mechanical damage, facilitation of photosynthetic carbon gain and carbon processing is enhanced by plasticity in plant form and microsite preference, enabling seedling survival and sapling growth inside and through the often severe boundary layer just above the ground cover. Several forms of facilitation (inanimate, interspecific, intraspecific and structural) result in substantial increases in photosynthetic carbon gain throughout the summer growth period, leading to enhanced root growth, subsequent amelioration of drought stress, and increased seedling survival. Avoidance of low temperatures and low-temperature photoinhibition of photosynthesis may be major benefits of the facilitation, enhancing photosynthetic carbon gain and respiratory-driven growth processes. We propose that the growth of vertical stems (flagged tree forms) from krummholz mats is analogous functionally to the facilitated growth of a seedling/sapling in and away from ground cover. Increasing abundance and growth of newly established trees in the treeline ecotone generates a structural and microsite facilitation characteristic of the subalpine forest below. This is followed by the formation of new subalpine forest with forest-like trees, and a new timberline at higher altitude. PMID- 14522717 TI - Ecophysiology of riparian cottonwoods: stream flow dependency, water relations and restoration. AB - Cottonwoods (Populus spp.) are adapted to riparian or floodplain zones throughout the Northern Hemisphere; they are also used as parents for fast-growing hybrid poplars. We review recent ecophysiological studies of the native cottonwoods Populus angustifolia James, P. balsamifera L., P. deltoides Marsh., P. fremontii S. Watson and P. trichocarpa T. & G. in North America, and P. nigra L. in Europe. Variation exists within and across species and hybrids; however, all riparian cottonwoods are dependent on shallow alluvial groundwater that is linked to stream water, particularly in semi-arid regions. This conclusion is based on studies of their natural occurrence, decline following river damming and dewatering (water removal), water relations, isotopic composition of xylem water, and by the establishment of cottonwoods along formerly barren natural channels after flow augmentation in response to the conveyance of irrigation water. When alluvial groundwater is depleted as a result of river dewatering or groundwater pumping, riparian cottonwoods exhibit drought-stress responses including stomatal closure and reduced transpiration and photosynthesis, altered 13C composition, reduced predawn and midday water potentials, and xylem cavitation. These physiological responses are accompanied by morphological responses including reduced shoot growth, altered root growth, branch sacrifice and crown die-back. In severe cases, mortality occurs. For example, severe dewatering of channels of the braided Big Lost River in Idaho led to mortality of the narrowleaf cottonwood, P. angustifolia, and adjacent sandbar willows, Salix exigua Nutt., within 5 years, whereas riparian woodlands thrived along flowing channels nearby. The conservation and restoration of cottonwoods will rely on the provision of river flow regimes that satisfy these ecophysiological requirements for survival, growth and reproduction. PMID- 14522718 TI - Diurnal and seasonal changes in stem increment and water use by yellow poplar trees in response to environmental stress. AB - To evaluate indicators of whole-tree physiological responses to climate stress, we determined seasonal, daily and diurnal patterns of growth and water use in 10 yellow poplar (Liriodendron tulipifera L.) trees in a stand recently released from competition. Precise measurements of stem increment and sap flow made with automated electronic dendrometers and thermal dissipation probes, respectively, indicated close temporal linkages between water use and patterns of stem shrinkage and swelling during daily cycles of water depletion and recharge of extensible outer-stem tissues. These cycles also determined net daily basal area increment. Multivariate regression models based on a 123-day data series showed that daily diameter increments were related negatively to vapor pressure deficit (VPD), but positively to precipitation and temperature. The same model form with slight changes in coefficients yielded coefficients of determination of about 0.62 (0.57-0.66) across data subsets that included widely variable growth rates and VPDs. Model R2 was improved to 0.75 by using 3-day running mean daily growth data. Rapid recovery of stem diameter growth following short-term, diurnal reductions in VPD indicated that water stored in extensible stem tissues was part of a fast recharge system that limited hydration changes in the cambial zone during periods of water stress. There were substantial differences in the seasonal dynamics of growth among individual trees, and analyses indicated that faster-growing trees were more positively affected by precipitation, solar irradiance and temperature and more negatively affected by high VPD than slower growing trees. There were no negative effects of ozone on daily growth rates in a year of low ozone concentrations. PMID- 14522719 TI - Photosynthate distribution patterns in cherrybark oak seedling sprouts. AB - We used 14C tracers to determine photosynthate distribution in cherrybark oak (Quercus pagoda Raf.) seedling sprouts following release from competing mid-story vegetation. Fall acquisition of labeled photosynthates by seedlings followed expected source-sink patterns, with root and basal stem tissues serving as the primary sinks. Four months after the seedlings had been labeled with 14C, they were clipped to induce sprouting. First-flush stem and leaf tissues of the resulting seedling sprouts were the primary sinks for labeled photosynthates stored in root tissues. Second-flush stem and leaf tissues, and first-flush stem and leaf tissues the following growing season, were not primary sinks for labeled photosynthates stored in root tissues despite the high radioactivity in root tissues. Root tissues appeared to deposit photosynthates in a layering process whereby the last photosynthates stored in new xylem were the first to be depleted during the initiation of a growth flush the following spring. There were more labeled photosynthates in roots of released seedling sprouts compared with non released seedling sprouts, indicating increased vigor of released seedling sprouts in response to greater light availability. In contrast, stem and source leaf tissues of non-released seedling sprouts contained greater percentages of labeled photosynthates compared with released seedling sprouts, indicating either greater sink strength or poorly developed xylem and phloem pathways that created inefficiencies in distribution to root tissues. The 14C distribution coefficients confirmed the distribution patterns and provided additional information on the important sinks in released and non-released cherrybark oak seedling sprouts. PMID- 14522720 TI - Effects of desiccation on the physiology and biochemistry of Quercus alba acorns. AB - Seeds that lose viability when dried to a water content of less than 12% are said to be recalcitrant. We subjected acorns of Quercus alba L., a species with recalcitrant seeds, to desiccation to determine the effects of drying on lipids, proteins and carbohydrates of the embryonic axis and cotyledon tissues. Samples of fresh seed and seed dried for selected intervals were analyzed for water content and germination, and for lipids, proteins and carbohydrates by Fourier transform-infrared (FT-IR) spectroscopy. Carbohydrates were further analyzed by gas chromatography (GC). The FT-IR analysis revealed that membrane lipid structure initially exhibited reversible shifts between gel and liquid crystalline phases in response to drying and rehydration; however, reversibility declined as viability was lost. Changes in carbohydrate concentration were observed based on peak height comparisons; sucrose concentration in the embryonic axis increased dramatically after 5 days of drying. The most sensitive indicator of desiccation damage was the irreversible change in protein secondary structure in embryonic axes and cotyledon tissue. These changes were illustrated by shifts in amide absorbance near 1650 cm(-1). Gas chromatography indicated an abundance of sucrose in both the embryonic axes and the cotyledon tissue. Although sucrose concentrations in these tissues were initially similar, sucrose concentration in the embryonic axes became significantly greater than in the cotyledons as the acorns dried. We hypothesize that, in drying acorns, increased concentration of sucrose does not prevent loss of viability, but acts as a glycoprotectant against cell collapse and cell wall membrane damage as water stress increases. PMID- 14522721 TI - But how much physical activity? PMID- 14522722 TI - A new look at dietary protein in diabetes. PMID- 14522723 TI - New evidence that iron supplementation during pregnancy improves birth weight: new scientific questions. PMID- 14522724 TI - The intestine and its microflora are partners for the protection of the host: report on the Danone Symposium "The Intelligent Intestine," held in Paris, June 14, 2002. AB - The intestine is an extremely complex living system that participates in the protection of the host through a strong defense against aggressions from the external environment. This defensive task is based on 3 constituents that are in permanent contact and dialog with each other: the microflora, mucosal barrier, and local immune system. We review herein current knowledge about these important functions. The gut microflora play a major role against exogenous bacteria through colonization resistance, but the mechanism of action is not yet established, although it is linked to the bacteria colonizing the gut. This colonization involves bacteria-bacteria dialog, bacteria-mucins interactions, and bacteria-colonocytes cross-talk associated with environmental factors. The intestinal mucosa is a cellular barrier and the main site of interaction with foreign substances and exogenous microorganisms. It is a complex physicochemical structure consisting of a mucous layer linked to cellular and stromal components that participate in the defense of the host through mucosal blood flow, mucosal secretions, epithelial cell functionals, surface hydrophobicity, and defensin production. The intestine is the primary immune organ of the body represented by the gut-associated lymphoid tissue through innate and acquired immunity. This immune system can tolerate dietary antigens and the gut-colonizing bacteria and recognizes and rejects enteropathogenic microorganisms that may challenge the body's defenses. In cooperation with these endogenous barriers, some in-transit bacteria, such as probiotics, can act as partners of the defense system of the intestine. PMID- 14522725 TI - Physical activity and weight loss: does prescribing higher physical activity goals improve outcome? AB - BACKGROUND: Recommending increased physical activity facilitates long-term weight loss, but the optimal level of physical activity to recommend is unknown. OBJECTIVE: The objective of the study was to evaluate the efficacy for long-term weight loss of recommendations for much higher physical activity than those normally used in behavioral treatments. DESIGN: Overweight men and women (n = 202) were randomly assigned to either a standard behavior therapy (SBT) for obesity, incorporating an energy expenditure (EE) goal of 1000 kcal/wk, or to a high physical activity (HPA) treatment, in which the goal was an EE of 2500 kcal/wk. To help HPA treatment group participants achieve this high exercise goal, their treatment included encouragement to recruit 1-3 exercise partners into the study, personal counseling from an exercise coach, and small monetary incentives. RESULTS: The HPA treatment group reported achieving higher mean (+/- SD) physical activity levels than did the SBT group at 6 mo (EE of 2399 +/- 1571 kcal/wk compared with 1837 +/- 1431 kcal/wk), 12 mo (EE of 2249 +/- 1751 kcal/wk compared with 1565 +/- 1309 kcal/wk), and 18 mo (EE of 2317 +/- 1854 kcal/wk compared with 1629 +/- 1483 kcal/wk) (all P < 0.01). Mean (+/- SEM) cumulative weight losses at 6, 12, and 18 mo in the HPA treatment group were 9.0 +/- 7.1, 8.5 +/- 7.9, and 6.7 +/- 8.1 kg, respectively. In the SBT group, the corresponding weight losses were 8.1 +/- 7.4, 6.1 +/- 8.8, and 4.1 +/- 7.3 kg, respectively. Between-group differences in weight loss were significant at 12 and 18 mo. CONCLUSION: These results suggest that recommendations of higher levels of physical activity (EE of 2500 kcal/wk) promote long-term weight loss better than do conventional recommendations. PMID- 14522726 TI - A case-control study of the association of diet and obesity with gout in Taiwan. AB - BACKGROUND: Gout has been a significant metabolic disorder for Chinese men in Taiwan; however, there is insufficient information on diet and lifestyle risk factors in this population. OBJECTIVE: The purpose of this case-control study was to explore potential dietary and lifestyle risk factors associated with gout in Chinese men. DESIGN: Between 1998 and 1999, we recruited and conducted face-to face interviews with patients from outpatient clinics in Taipei who had incident gout (n = 92) and with their healthy coworkers (controls; n = 92). RESULTS: Systolic blood pressure, diastolic blood pressure, waist-to-hip ratio, waist-to height ratio, and body mass index were significantly higher in cases than in controls. Family histories of gout and diabetes mellitus were strong risk factors for gout. Frequencies of vegetable and fruit consumption were significantly lower in cases than in controls. Logistic regression analyses showed that high alcohol intake and low intakes of fiber, folate, and vitamin C increased the risk of gout, but no association was found with purine intake. After covariates were controlled for, the adjusted odds ratios for the middle and highest tertiles of waist-to-height ratio (0.50-0.54 and >/==" BORDER="0"> 0.55, respectively) were 3.89 (95% CI: 1.32, 11.46) and 4.37 (1.18, 16.22), respectively, but no linear association was found for waist-to-hip ratio and waist circumference. CONCLUSIONS: Consumption of alcohol, but not of purine, may be a significant dietary risk factor for gout. Food sources rich in dietary fiber, folate, and vitamin C, such as fruit and vegetables, protect against gout. Waist-to-height ratio, which indicates central obesity, has a significant linear effect on gout occurrence, independent of body mass index. PMID- 14522727 TI - Incorporation of docosahexaenoic acid into nerve membrane phospholipids: bridging the gap between animals and cultured cells. AB - BACKGROUND: Functional maturation of nervous tissues depends on membrane accretion of docosahexaenoic acid (DHA). Animal studies have shown that incorporation of dietary DHA into membrane phospholipids is dose dependent. The molecular effects of DHA are commonly studied in cultured cells, but questions remain about the physiologic connection between animal and cell models. OBJECTIVE: We developed a linear model for comparing the responses of rat nervous tissues to dietary DHA with the responses of human cell lines to DHA in medium. DESIGN: Rats were rendered chronically deficient in n-3 fatty acids by being reared on a peanut oil diet. DHA status was replenished in the F2 generation by using increasing supplements of a microalgal oil. Human retinoblastoma and neuroblastoma cells were dosed with unesterified DHA. DHA accumulation into phospholipids was defined by the plateau of the dose-response curve (DHA(max)) and by the supplement required to produce one-half the DHA(max) (DHA(50)). RESULTS: The DHA(max) values for 4 brain regions and 2 neuroblastoma lines were similar, and the value for the retinoblastoma line was similar to the retinal value. Expressing the DHA input as micro mol/10 g diet and as micro mol/L medium resulted in similar values for the ratio of DHA(max) to DHA(50) in the 4 brain regions and the 3 cell lines. The DHA(max)-DHA(50) ratios in the ethanolamine phosphoglyceride and phosphatidylcholine fractions in retinal phospholipids were 6 and 10 times, respectively, those in the brain and cultured cells. CONCLUSIONS: The dose-dependent responses of cells and the brain to DHA supplements can be compared by using DHA(max)-DHA(50) ratios. We propose a counting frame that allows the comparison of the dose responses of the brain and cells to exogenous DHA. PMID- 14522728 TI - Randomized controlled crossover study of the effect of a highly beta-glucan enriched barley on cardiovascular disease risk factors in mildly hypercholesterolemic men. AB - BACKGROUND: Soluble-fiber beta-glucan derived from oats can reduce cardiovascular disease (CVD) risk through reductions in total and LDL cholesterol. Barley derived beta-glucan may also improve serum cholesterol, but large quantities are required for clinical significance. OBJECTIVE: This trial investigated whether a beta-glucan-enriched form of barley can favorably modify cholesterol and other markers of CVD and diabetes risk. DESIGN: Eighteen mildly hyperlipidemic ( +/- SD: 4.0 +/- 0.6 mmol LDL cholesterol/L) men with a mean (+/- SD) body mass index (in kg/m(2)) of 27.4 +/- 4.6 were randomly assigned in this single-blind, 2 x 4 wk trial to either the treatment arm [8.1-11.9 g beta-glucan/d (scaled to body weight)] or the control arm (isoenergetic dose of 6.5-9.2 g glucose/d). After a washout period of 4 wk, dietary regimens were crossed over. The trial took place in a long-stay metabolic facility, and all foods were provided (38% of energy from fat). Fasted blood samples were collected on days 0, 1, 7, 14, 21, 28, and 29 in both study arms. An oral-glucose-tolerance test was carried out on days 0 and 29. RESULTS: There was no significant change (Delta) in total (Delta = -0.08 mmol/L, -1.3%), LDL (Delta = -0.15 mmol/L, -3.8%), or HDL (Delta = 0 mmol/L) cholesterol or in triacylglycerol (Delta = 0.18 mmol/L), fasting glucose (Delta = -0.05 mmol/L), or postprandial glucose when analyzed between treatments (P > 0.05; ANOVA). CONCLUSION: The effect of beta-glucan-enriched barley on lipid profile was highly variable between subjects, and there was no evidence of a clinically significant improvement in CVD risk across this group of mildly hyperlipidemic men. PMID- 14522729 TI - Prospective study of the association of changes in dietary intake, physical activity, alcohol consumption, and smoking with 9-y gain in waist circumference among 16 587 US men. AB - BACKGROUND: Although it is known that abdominal obesity increases the risk of chronic diseases, prospective data examining the relation between lifestyle factors and the accumulation of abdominal adipose tissue are sparse. OBJECTIVE: The objective of the study was to determine the associations of changes in diet, physical activity, alcohol consumption, and smoking with 9-y waist gain among US men. DESIGN: A prospective cohort comprised 16 587 US men aged 40-75 y at baseline in 1986. Data on lifestyle factors were provided periodically with the use of self-reported questionnaires, and participants measured and reported their waist circumference in 1987 and 1996. RESULTS: In multivariate analyses, a 2% increment in energy intake from trans fats that were isocalorically substituted for either polyunsaturated fats or carbohydrates was significantly associated with a 0.77-cm waist gain over 9 y (P < 0.001 for each comparison). An increase of 12 g total fiber/d was associated with a 0.63-cm decrease in waist circumference (P < 0.001), whereas smoking cessation and a 20-h/wk increase in television watching were associated with a 1.98-cm and 0.59-cm waist gain, respectively (P < 0.001). Increases of 25 metabolic equivalent tasks (METs) * h/wk in vigorous physical activity and of >/= 0.5 h/wk in weight training were associated with 0.38-cm and 0.91-cm decreases in waist circumference, respectively (P < 0.001 for each comparison). These associations remained significant after further adjustment for concurrent change in body mass index. Changes in total fat and alcohol consumption and in walking volume were not significantly related to waist gain. CONCLUSIONS: Waist gain may be modulated by changes in trans fat and fiber consumption, smoking cessation, and physical activity. PMID- 14522730 TI - Coffee acutely modifies gastrointestinal hormone secretion and glucose tolerance in humans: glycemic effects of chlorogenic acid and caffeine. AB - BACKGROUND: Accumulating evidence suggests that certain dietary polyphenols have biological effects in the small intestine that alter the pattern of glucose uptake. Their effects, however, on glucose tolerance in humans are unknown. OBJECTIVE: The objective was to investigate whether chlorogenic acids in coffee modulate glucose uptake and gastrointestinal hormone and insulin secretion in humans. DESIGN: In a 3-way, randomized, crossover study, 9 healthy fasted volunteers consumed 25 g glucose in either 400 mL water (control) or 400 mL caffeinated or decaffeinated coffee (equivalent to 2.5 mmol chlorogenic acid/L). Blood samples were taken frequently over the following 3 h. RESULTS: Glucose and insulin concentrations tended to be higher in the first 30 min after caffeinated coffee consumption than after consumption of decaffeinated coffee or the control (P < 0.05 for total and incremental area under the curve for glucose and insulin). Glucose-dependent insulinotropic polypeptide secretion decreased throughout the experimental period (P < 0.005), and glucagon-like peptide 1 secretion increased 0-120 min postprandially (P < 0.01) after decaffeinated coffee consumption compared with the control. Glucose and insulin profiles were consistent with the known metabolic effects of caffeine. However, the gastrointestinal hormone profiles were consistent with delayed intestinal glucose absorption. CONCLUSIONS: Differences in plasma glucose, insulin, and gastrointestinal hormone profiles further confirm the potent biological action of caffeine and suggest that chlorogenic acid might have an antagonistic effect on glucose transport. Therefore, a novel function of some dietary phenols in humans may be to attenuate intestinal glucose absorption rates and shift the site of glucose absorption to more distal parts of the intestine. PMID- 14522731 TI - An increase in dietary protein improves the blood glucose response in persons with type 2 diabetes. AB - BACKGROUND: In single-meal studies, dietary protein does not result in an increase in glucose concentrations in persons with or without type 2 diabetes, even though the resulting amino acids can be used for gluconeogenesis. OBJECTIVE: The metabolic effects of a high-protein diet were compared with those of the prototypical healthy (control) diet, which is currently recommended by several scientific organizations. DESIGN: The metabolic effects of both diets, consumed for 5 wk each (separated by a 2-5-wk washout period), were studied in 12 subjects with untreated type 2 diabetes. The ratio of protein to carbohydrate to fat was 30:40:30 in the high-protein diet and 15:55:30 in the control diet. The subjects remained weight-stable during the study. RESULTS: With the fasting glucose concentration used as a baseline from which to determine the area under the curve, the high-protein diet resulted in a 40% decrease in the mean 24-h integrated glucose area response. Glycated hemoglobin decreased 0.8% and 0.3% after 5 wk of the high-protein and control diets, respectively; the difference was significant (P < 0.05). The rate of change over time was also significantly greater after the high-protein diet than after the control diet (P < 0.001). Fasting triacylglycerol was significantly lower after the high-protein diet than after the control diet. Insulin, C-peptide, and free fatty acid concentrations were not significantly different after the 2 diets. CONCLUSION: A high-protein diet lowers blood glucose postprandially in persons with type 2 diabetes and improves overall glucose control. However, longer-term studies are necessary to determine the total magnitude of response, possible adverse effects, and the long term acceptability of the diet. PMID- 14522732 TI - Different glycemic indexes of breakfast cereals are not due to glucose entry into blood but to glucose removal by tissue. AB - BACKGROUND: The glycemic index (GI) of a food is thought to directly reflect the rate of digestion and entry of glucose into the systemic circulation. The blood glucose concentration, however, represents a balance of both the entry and the removal of glucose into and from the blood, respectively. Such direct quantification of the postprandial glucose curve with respect to interpreting the GI is lacking in the literature. OBJECTIVE: We compared the plasma glucose kinetics of low- and high-GI breakfast cereals. DESIGN: On 2 occasions, plasma insulin concentrations and plasma glucose kinetics (by constant-rate infusion of [6,6-(2)H(2)]glucose) were measured in 6 healthy males for 180 min after they fasted overnight and then consumed an amount of corn flakes (CF) or bran cereal (BC) containing 50 g available carbohydrate. RESULTS: The GI of CF was more than twice that of BC (131.5 +/- 33.0 compared with 54.5 +/- 7.2; P < 0.05), despite no significant differences in the rate of appearance of glucose into the plasma during the 180-min period. Postprandial hyperinsulinemia occurred earlier with BC than with CF, resulting in a 76% higher plasma insulin concentration at 20 min (20.4 +/- 4.5 compared with 11.6 +/- 2.1 micro U/mL; P < 0.05). This was associated with a 31% higher rate of disappearance of glucose with BC than with CF during the 30-60-min period (28.7 +/- 3.1 compared with 21.9 +/- 3.1 micro mol. kg(-)(1). min(-)(1); P < 0.05). CONCLUSION: The lower GI of BC than of CF was not due to a lower rate of appearance of glucose but instead to an earlier postprandial hyperinsulinemia and an earlier increase in the rate of disappearance of glucose, which attenuated the increase in the plasma glucose concentration. PMID- 14522733 TI - Differential utilization of systemic and enteral ammonia for urea synthesis in control subjects and ornithine transcarbamylase deficiency carriers. AB - BACKGROUND: Female carriers of ornithine transcarbamylase deficiency (OTCD) have nearly normal rates of total urea synthesis, but they derive less urea from systemic glutamine amide nitrogen than do healthy persons. OBJECTIVE: The objective of the study was to investigate whether females with symptomatic OTCD rely on alternative pathways to compensate for the reduced urea synthesis activity observed in this disorder. DESIGN: The 5-d study involved 6 control subjects (3 males, 3 females) and 6 female OTCD carriers who had a fixed energy intake of 133 kJ. kg(-)(1). d(-)(1) and a protein intake of 0.8 g. kg(-)(1). d( )(1). They underwent two 12-h periods of isotopic tracer administration, separated by 2 d. On both occasions, [(18)O] or [(13)C]urea was infused intravenously, and the subjects consumed hourly meals. During the first period, [(15)N]NH(4)Cl was given intravenously; during the second period, the tracer was given as hourly oral doses. RESULTS: OTCD carriers produced less urea (P < 0.05) but had a higher (P < 0.05) mean ammonia appearance rate and plasma ammonia concentration than did control subjects. OTCD carriers incorporated a lower (P < 0.001) mean (+/- SE) proportion of the intravenous [(15)N]NH(4)Cl dose into circulating urea than did control subjects (16 +/- 1% compared with 36 +/- 2%), but there was no genotypic difference in the incorporation of orally administered tracer (81 +/- 4% compared with 72 +/- 4%, respectively). CONCLUSION: A good degree of dietary protein tolerance seemed to be retained in OTCD carriers by the maintenance of higher ammonia appearance rates, expansion of the plasma ammonia pool, and reliance on the ability of the perivenous hepatocytes to clear excess ammonia via glutamine synthesis. PMID- 14522734 TI - Suggested lower cutoffs of serum zinc concentrations for assessing zinc status: reanalysis of the second National Health and Nutrition Examination Survey data (1976-1980). AB - BACKGROUND: The risk of zinc deficiency in populations can be estimated by comparing serum zinc data with statistically defined lower cutoffs derived from a presumably healthy population. Serum zinc data are available from a large sample of the US population assessed during the second National Health and Nutrition Examination Survey (NHANES II). Although the original analysis of these data considered fasting status and the time of day of blood sampling, it did not account for potentially confounding variables that may affect the serum zinc concentration, such as age, sex, and health status. OBJECTIVE: The objective was to describe variations in serum zinc concentration by age, sex, and other characteristics and to recommend lower cutoffs for presumably healthy persons. DESIGN: Serum zinc data from NHANES II were analyzed by using analysis of variance and covariance models to identify and describe variables significantly associated with serum zinc concentration; 2.5th percentile curves were produced and used to establish age- and sex-based lower cutoffs. RESULTS: Age and sex were significant confounders of serum zinc concentration, so separate lower cutoffs were derived for children and adolescent and adult males and females. Other minor confounding variables were identified. Tentative lower cutoffs for pregnancy and oral contraceptive use were also derived. CONCLUSIONS: The interpretation of population serum zinc data with the use of lower cutoffs should account for the age and sex of the subjects, pregnancy and oral contraceptive use, and fasting status and time of day of blood collection. PMID- 14522735 TI - B vitamin status and concentrations of homocysteine and methylmalonic acid in elderly German women. AB - BACKGROUND: Prior investigations found that elderly persons are at higher risk than are younger persons for B vitamin deficiency, which leads to elevated plasma total homocysteine (tHcy) concentrations that are associated with an increased risk for certain diseases such as coronary artery disease. To date, published data have shown decreased vitamin status and elevated tHcy among the elderly. OBJECTIVE: We evaluated the dietary intake and the blood status of various B vitamins and tHcy and methylmalonic acid (MMA) concentrations in 178 younger (60 70-y-old) female seniors. DESIGN: Dietary intake was assessed with a 3-d diet record. Thiamine, riboflavin, and vitamin B-6 activity coefficients of erythrocyte transketolase (EC 2.2.1.1), erythrocyte glutathione reductase (EC 1.6.4.2), and erythrocyte alpha-aspartic aminotransferase (EC 2.6.1.1) were used as functional indexes for the status of the 3 vitamins, respectively. Concentrations of serum and red blood cell folate, serum cobalamin and MMA, and plasma tHcy were measured. RESULTS: Indexes of thiamine, pyridoxine, and cobalamin indicated insufficient status in one-third of the women, whereas tHcy and MMA concentrations were elevated in 17.4% and 9.6% of the women, respectively. An association between vitamin intake and vitamin concentration in the blood was found only for folate. The mean tHcy concentration in subjects in the lowest quartile of serum folate concentration was 23% higher than that in subjects in the highest quartile. There was no association between riboflavin and tHcy concentrations. MMA was positively correlated with age and inversely correlated with serum cobalamin concentration. CONCLUSIONS: Even in younger, well educated, female seniors, the prevalence of low B vitamin status and elevated plasma tHcy concentration is high. Thiamine, pyridoxine, folate, and cobalamin supplementation should be considered. PMID- 14522736 TI - Iron supplementation during pregnancy, anemia, and birth weight: a randomized controlled trial. AB - BACKGROUND: The need for prophylactic iron during pregnancy is uncertain. OBJECTIVE: We tested the hypothesis that administration of a daily iron supplement from enrollment to 28 wk of gestation to initially iron-replete, nonanemic pregnant women would reduce the prevalence of anemia at 28 wk and increase birth weight. DESIGN: Between June 1995 and September 1998, 513 low income pregnant women in Cleveland were enrolled in the study before 20 wk of gestation. Of these, 275 had a hemoglobin concentration >/= 110 g/L and a ferritin concentration >/= 20 micro g/L and were randomly assigned to receive a monthly supply of capsules containing either 30 mg Fe as ferrous sulfate or placebo until 28 wk of gestation. At 28 and 38 wk of gestation, women with a ferritin concentration of 12 to < 20 micro g/L or < 12 micro g/L received 30 and 60 mg Fe/d, respectively, regardless of initial assignment. Almost all the women received some supplemental iron during pregnancy. We obtained infant birth weight and gestational age at delivery for 117 and 96 of the 146 and 129 women randomly assigned to receive iron and placebo, respectively. RESULTS: Compared with placebo, iron supplementation from enrollment to 28 wk of gestation did not significantly affect the overall prevalence of anemia or the incidence of preterm births but led to a significantly higher mean (+/- SD) birth weight (206 +/- 565 g; P = 0.010), a significantly lower incidence of low-birth-weight infants (4% compared with 17%; P = 0.003), and a significantly lower incidence of preterm low birth-weight infants (3% compared with 10%; P = 0.017). CONCLUSION: Prenatal prophylactic iron supplementation deserves further examination as a measure to improve birth weight and potentially reduce health care costs. PMID- 14522737 TI - Milk folate but not milk iron concentrations may be inadequate for some infants in a rural farming community in San Mateo, Capulhuac, Mexico. AB - BACKGROUND: We were interested in identifying possible nutritional reasons for growth faltering among breastfed infants in the rural farming community of San Mateo, Capulhuac, Mexico (2800 m above sea level). OBJECTIVE: We examined the prevalence of inadequate iron and folate status among lactating Otomi women and determined to what extent their iron and folate nutriture influenced the milk concentrations of these nutrients. DESIGN: Lactating women (n = 71) provided blood and milk samples and dietary information at a mean (+/- SD) of 22 +/- 13 d postpartum. Blood indexes included hemoglobin, hematocrit, serum iron, total-iron binding capacity, ferritin, transferrin receptor, mean cell volume, plasma folate, and erythrocyte folate. RESULTS: Approximately 62% and 58% of the women had nutritional anemia defined as a hemoglobin concentration /= 65 y with acute myocardial infarction by evaluating BMI and major (death and cardiogenic shock) and minor in-hospital outcomes. RESULTS: In hospital death, cardiogenic shock, and the combined endpoints were not significantly different between elderly patients with and without preinfarction angina. Interestingly, in-hospital death, cardiogenic shock, and the combined endpoints were significantly fewer in elderly patients with than without preinfarction angina in the subset of patients with the lowest BMI (P < 0.01, < 0.01, and < 0.01, respectively). Regression analysis showed that preinfarction angina did not protect against in-hospital death when analyzed in all patients independently of BMI, whereas it was protective in the subset of patients with the lowest BMI (odds ratio: 0.06; 95% CI: 0.00, 0.54). CONCLUSIONS: Preinfarction angina does not protect against in-hospital death, cardiogenic shock, or the combined endpoints in elderly patients with acute myocardial infarction. With stratification by quartiles of BMI, the protective effect of preinfarction angina is preserved in elderly patients with the lowest BMI. PMID- 14522740 TI - Paleolithic diet, sweet potato eaters, and potential renal acid load. PMID- 14522742 TI - Metabolic effects of dietary fructose. PMID- 14522745 TI - Defining and interpreting intakes of sugars. AB - This paper clarifies the myriad of terminologies used to describe intakes of sugars by American consumers. In addition, it carefully critiques information sources used to explain and interpret consumption levels. Sugars are incorporated into foods for their biological, sensory, physical, and chemical properties. By chemical definition, the sugars normally consumed are the monosaccharides and disaccharides: glucose, fructose, galactose, sucrose, lactose, maltose, and trehalose. US governmental agencies use 4 terms to describe sugars: added sugars, caloric sweeteners, sugar, and sugars. Different sources are included when measuring sugars. Knowledge regarding intakes of sugars relies on food intake surveys (primarily dietary recalls) and economic food availability estimates. Although intake data may underestimate actual consumption, availability data tend to overestimate it. Furthermore, the sugars contents of many foods appearing in composition databases are derived from the summation of recipe ingredients rather than from actual measurements. Intakes of sugars over time (trends) must be viewed within the context of varying definitions, changes in food composition, changes in dietary intake methods, and acknowledged increases in the underreporting of intake. Agreement is needed to identify one common definition to describe intakes of sugars. Convergence between intake data and economic availability data would more accurately depict consumption. Precise amounts of sugars within currently available foods should be measured, not calculated. Without a common language, accurate and precise measurements, and consensus among scientists, educators, regulatory agencies, and the public, conversations regarding any health effects of sugars may lead to continued misunderstandings. PMID- 14522746 TI - The scientific basis of recent US guidance on sugars intake. AB - Most fruits and dairy products are high in sugars, and thus naturally occurring sugars are consumed as part of a healthy diet. Sugars are also added to foods during processing or preparation, primarily to enhance taste. Monosaccharides and disaccharides added to foods are chemically indistinguishable from naturally occurring forms. However, concern has been expressed about the apparent increasing consumption of added sugars and their possible role in displacing or diluting nutrients in the diet and contributing to the epidemic of obesity in developed countries. One of the 2000 Dietary Guidelines for Americans states, "Choose beverages and foods to moderate your intake of sugars." The Dietary Guidelines Advisory Committee discussed whether to specify added sugars rather than the broader term sugars but decided that it was not possible to conclude that added sugars per se play a negative role in the public's health. The Dietary Guidelines Advisory Committee issued a call for more definitive studies of the role of sugars in current diets and the potential effect of a reduction in added sugars on both dietary quality and energy intake. The American Heart Association recently released a statement advising consumers to limit sugars consumption. The macronutrient report for the dietary reference intakes addresses many of these same issues; the expert panel concluded that it was not appropriate to set a tolerable upper intake level for added sugars but suggested a maximal intake level of 25% of energy from added sugars because of concerns about reduced intakes of essential micronutrients. PMID- 14522748 TI - Consumption of sugars and the regulation of short-term satiety and food intake. AB - This review examines the relation between the consumption of sugars and their effects on short-term (ie, to 2 h) satiety and food intake in humans. Many factors need to be considered in the evaluation of reported studies and the conclusions derived from this body of literature. These factors include evaluation of the dose and form (solid or liquid) of the treatments, time of day administered, characteristics of the subjects, sample size, and approaches used to measure satiety and food intake. Mechanisms by which sugars may signal regulatory systems for food intake need to be considered when evaluating both study designs and conclusions. For this reason, the relation between the blood glucose response to sugar consumption and subsequent feeding behavior is also examined. It is concluded that sugars stimulate satiety mechanisms and reduce food intake in the short term and that the mechanisms by which this response occurs cannot be attributed solely to their effect on blood glucose. PMID- 14522747 TI - Sugars: hedonic aspects, neuroregulation, and energy balance. AB - The prevalence of obesity has increased dramatically in recent years in the United States, with similar patterns seen in several other countries. Although there are several potential explanations for this dramatic increase in obesity, dietary influences are a contributing factor. An inverse correlation between dietary sugar intake and body mass index has been reported, suggesting beneficial effects of carbohydrate intake on body mass index. In this review we discuss how sugars interact with regulatory neurochemicals in the brain to affect both energy intake and energy expenditure. These neurochemicals appear to be involved in dietary selection, and sugars and palatable substances affect neurochemical changes in the brain. For example, rats that drink sucrose solutions for 3 wk have major changes in neuronal activity in the limbic area of the brain, a region involved in pleasure and other emotions. We also investigate the relations between sucrose (and other sweet substances), drugs of abuse, and the mesolimbic dopaminergic system. The presence of sucrose in an animal's cage can affect the animals desire to self-administer drugs of abuse. Also, an animal's level of sucrose preference can predict its desire to self-administer cocaine. Such data suggest a relation between sweet taste and drug reward, although the relevance to humans is unclear. Finally, we address the influence of sugar on body weight control. For example, sucrose feeding for 2 wk decreases the efficiency of energy utilization and increases gene expression of uncoupling protein 3 in muscle, suggesting that sucrose may influence uncoupling protein 3 activity and contribute to changes in metabolic efficiency and thus regulation of body weight. PMID- 14522749 TI - Sugars, energy metabolism, and body weight control. AB - Obesity represents a major threat to health and quality of life. Although obesity has strong genetic determinants, it is generally accepted that it results from an imbalance between food intake and daily physical activity. Health guidelines have been focused on 3 particular lifestyle factors: increased levels of physical activity and reductions in the intakes of fat and sugars. The dietary guidelines, especially, are under debate. This review covers evidence from carefully controlled laboratory studies, clinical trials, studies in populations at high risk of developing obesity, and epidemiologic studies on the role of sugars, particularly sucrose, in the development of obesity. Although many environmental factors promote a positive energy balance, it is clear that the consumption of a low-carbohydrate, high-fat diet increases the likelihood of weight gain. The evidence related to carbohydrate, particularly sugars, and the type of food (solid or liquid) is less clear because the number of long-term ad libitum dietary intervention trials is very small. Data on sucrose intake in relation to metabolism and weight gain do not associate high consumption of sucrose with the prevalence of obesity. The evidence supports the current dietary guidelines for reducing fat intake. However, the effect of the carbohydrate source and class and of the form in which carbohydrate is consumed (solid or liquid) on body weight control requires further consideration. PMID- 14522750 TI - Sugars and starch in the nutritional management of diabetes mellitus. AB - Nutritional recommendations, long recognized as an important aspect of diabetes mellitus treatment, have also been an area of persistent controversy, particularly regarding the proportions and types of carbohydrate and fat. This review addresses the role of sugars within medical nutrition therapy for diabetes mellitus. Nutritional recommendations for diabetes mellitus treatment were revised recently. The new guidelines do not specifically restrict intake of sugars, although general recommendations are made for including fiber, whole grains, vegetables, and fruits within dietary selections containing starches. For carbohydrates, the principle focus is on overall caloric amounts. In type 1 diabetes the most effective approach to the control of postprandial hyperglycemia continues to be adjustment of premeal doses of insulin on the basis of carbohydrate counting. In type 2 diabetes, in addition to a focus on caloric content of carbohydrate, consideration continues to be given to the role of the glycemic index as a determinant of postprandial hyperglycemia and overall metabolic control. Nevertheless, consensus recommendations do not support widespread use of the glycemic index. An area of some change is a more clear endorsement of including monounsaturated fatty acids. Current recommendations are that monounsaturated fatty acids and carbohydrates combined should provide 60-70% of daily energy intake, with individual flexibility in the respective proportions, whereas intake of saturated fats is limited to < 10% of energy intake. This new emphasis reflects greater awareness of the importance of responding to individual and cultural dietary preferences and the need to address treatment of both hyperglycemia and dyslipidemia in diabetes mellitus. PMID- 14522751 TI - Sugars, insulin sensitivity, and the postprandial state. AB - Since insulin resistance was first associated with type 2 diabetes and later with cardiovascular disease and hypertension, there has been considerable interest in the role of dietary and environmental factors. Sucrose and fructose have been a particular research focus. Research on animals, particularly rodents, has shown a clear and consistent effect of high-sucrose and high-fructose diets in decreasing insulin sensitivity. Experiments in humans have produced very conflicting results, with limited evidence for a negative effect on insulin sensitivity at higher intakes of fructose or sucrose (generally > 30% of daily energy from sucrose and > 15% of daily energy from fructose). Observation studies in humans have not shown a link between sucrose consumption and insulin sensitivity independent of other dietary factors. This is in contrast with several small studies that showed an improvement in insulin sensitivity after subjects followed dietary advice to lower the glycemic index of their food choices (where sugars were not a target for change). However, the pattern of postprandial glucose and insulin responses elicited by sucrose and fructose differs substantially from that elicited by starches, with lower troughs elicited by sucrose and fructose 2 3 h after eating. These differences in the pattern of postprandial responses offer a potential explanation for the conflicting results on insulin sensitivity, with the possibility that increases in insulin exposure may affect insulin sensitivity through down-regulation of insulin action. PMID- 14522752 TI - Sugars, hypertriglyceridemia, and cardiovascular disease. AB - Short-term studies consistently show that raising the carbohydrate content of the diet increases serum triacylglycerol concentrations. As compared with starches, sugars (particularly sucrose and fructose) tend to increase serum triacylglycerol concentrations by approximately 60%. The magnitude of the effect depends on other aspects of the diet, including the total amount of carbohydrate and the types of fat, carbohydrate, and fiber, but definitive studies to describe the dose response relations are not available. Longer-term studies show that some high carbohydrate diets are not associated with increased fasting serum triacylgycerol concentrations. However, sedentary subjects with upper-body and visceral obesity who have the metabolic syndrome tend to be at higher risk for hypertriglyceridemia in response to high-sucrose and high-carbohydrate diets; moderate weight loss mitigates the effect. Hyperinsulinemia or insulin resistance may play a role in promoting higher rates of VLDL synthesis and hypertriglyceridemia in obesity, but the mechanisms remain unclear. The effect of fructose in promoting triacylglycerol synthesis is independent of insulinemia, however. In terms of the long-term effects of diets high in sugars on the risk of cardiovascular disease, available epidemiologic evidence indicates no association of sugars or total carbohydrate intake per se, but high dietary glycemic load is associated with higher serum triacylglycerol concentrations and greater risk of coronary heart disease in women. Studies are needed to delineate the independent effects of dietary sugars and glycemic load on serum triacylglycerol concentrations in lean and obese men and women and to determine whether the elevations in fasting and fed concentrations of serum triacylglycerol with high carbohydrate and high-sugars diets are associated with increased risk of cardiovascular disease. PMID- 14522754 TI - Sugars and Health Workshop: summary and conclusions. PMID- 14522753 TI - Sugars and dental caries. AB - A dynamic relation exists between sugars and oral health. Diet affects the integrity of the teeth; quantity, pH, and composition of the saliva; and plaque pH. Sugars and other fermentable carbohydrates, after being hydrolyzed by salivary amylase, provide substrate for the actions of oral bacteria, which in turn lower plaque and salivary pH. The resultant action is the beginning of tooth demineralization. Consumed sugars are naturally occurring or are added. Many factors in addition to sugars affect the caries process, including the form of food or fluid, the duration of exposure, nutrient composition, sequence of eating, salivary flow, presence of buffers, and oral hygiene. Studies have confirmed the direct relation between intake of dietary sugars and dental caries across the life span. Since the introduction of fluoride, the incidence of caries worldwide has decreased, despite increases in sugars consumption. Other dietary factors (eg, the presence of buffers in dairy products; the use of sugarless chewing gum, particularly gum containing xylitol; and the consumption of sugars as part of meals rather than between meals) may reduce the risk of caries. The primary public health measures for reducing caries risk, from a nutrition perspective, are the consumption of a balanced diet and adherence to dietary guidelines and the dietary reference intakes; from a dental perspective, the primary public health measures are the use of topical fluorides and consumption of fluoridated water. PMID- 14522755 TI - Assessing the treatment of angle closure. PMID- 14522756 TI - Interventions for angle-closure glaucoma: an evidence-based update. AB - PURPOSE: To assess the interventions to treat acute angle closure (AAC) and primary angle closure (PAC) with or without glaucomatous optic neuropathy. CLINICAL RELEVANCE: Primary angle closure is one of the leading causes of blindness in East Asia. At present, there are few clinical guidelines on the optimal treatment of AAC or PAC in the affected or contralateral eye. METHODS: All randomized clinical trials, prospective controlled clinical trials, nonprospective controlled clinical trials, and retrospective case series with >50 cases that evaluated treatments for AAC or PAC were included. Studies published in the English language were identified from MEDLINE, PubMed, EMBASE, and the Cochrane Collaborations, as well as by a hand search of the reference lists of important articles. RESULTS: Nine randomized clinical trials and 24 nonrandomized clinical trials and large case series were evaluated. Laser peripheral iridotomy (LPI) has been found to be as effective as surgical peripheral iridectomy in randomized clinical trials of the affected and contralateral eyes of AAC or PAC patients with or without evidence of glaucoma. In another randomized clinical trial, latanoprost was found to decrease intraocular pressure (IOP) more than timolol for PAC in patients for whom LPI alone failed. CONCLUSIONS: This review suggests that LPI should be recommended for the treatment of affected and contralateral eyes of AAC patients. In patients with PAC and insufficient treatment with LPI, latanoprost eye drops may decrease IOP more than timolol. There is still insufficient evidence about other interventions for the treatment of AAC and PAC. PMID- 14522758 TI - Angle closure in younger patients. AB - PURPOSE: Angle-closure glaucoma is rare in children and young adults; only scattered cases associated with specific clinical entities have been reported. We evaluated the findings in patients aged 40 or younger with angle closure in our database. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Our database was searched for patients aged 40 years or younger with angle closure. Data recorded included age at initial consultation, age at the time of diagnosis, gender, slit-lamp examination, gonioscopy, ultrasound biomicroscopy (from 1993 onward), clinical diagnosis, and therapy. Patients with prior incisional surgery were excluded, as were patients with anterior chamber proliferative mechanisms leading to angle closure. RESULTS: Sixty-seven patients (49 females, 18 males) met entry criteria. Mean age at the time of consultation was 34.4 +/- 9.4 (standard deviation) years (range, 3-68 years). Diagnoses included plateau iris syndrome (35 patients); iridociliary cysts (8 patients); retinopathy of prematurity (7 patients); uveitis (5 patients); isolated nanophthalmos (3 patients); relative pupillary block (2 patients); Weill-Marchesani syndrome (3 patients); and 1 patient each with Marfan syndrome, miotic-induced angle closure, persistent hyperplastic primary vitreous, and idiopathic lens subluxation. CONCLUSIONS: The etiology of angle closure in young individuals differs from the older population and is typically associated with structural/developmental ocular anomalies rather than relative pupillary block. After laser iridotomy, these eyes should be monitored for recurrent angle closure and the need for additional laser or incisional surgical intervention. PMID- 14522759 TI - Patterns of visual field defects in chronic angle-closure glaucoma with different disease severity. AB - PURPOSE: To evaluate the patterns of visual field defects in patients with chronic angle-closure glaucoma (CACG) with varying extent of optic nerve damage. DESIGN: Prospective, consecutive, observational case series. PARTICIPANTS: One hundred forty-six Asian patients with well-controlled CACG. METHODS: Visual field tests were performed using program 24-2 of the Humphrey Field Analyzer (Humphrey Instruments, San Leandro, CA) with the Swedish interactive thresholding algorithm standard. One hundred ten eligible visual fields were scored with the system adopted by the Advanced Glaucoma Intervention Study and were categorized into 4 groups accordingly: mild, moderate, severe, and end-stage. Each hemifield was divided into the nasal, paracentral, and arcuate areas, and field loss that involved respective areas was defined as nasal step, paracentral scotoma, and arcuate scotoma. MAIN OUTCOME MEASURES: The distribution of field defect patterns in each group was evaluated. The mean deviation (MD) was compared among the 3 areas within one hemifield and between each pair of corresponding areas across the median raphe. RESULTS: The nasal area was the most commonly damaged area in the mild group, being noted in 52% of eyes in the superior hemifield and 58% of eyes in the inferior hemifield. In the moderate group, field loss involving both the nasal and arcuate areas dominated the superior hemifield, whereas field loss involving all three areas dominated the inferior hemifield. The MD of the nasal area was the worst among the three areas in each hemifield of the mild and moderate groups, as well as in the inferior hemifield of the severe group (all P < 0.001). There were no significant differences in the MD of each area between the superior hemifield and their inferior counterparts. However, the superior hemifield as a whole showed a better MD than the inferior hemifield (P=0.034) in the mild group. CONCLUSIONS: Visual field loss that involved the nasal area was the most common pattern in the early stage of CACG. The MD of the nasal area was worse than those of the arcuate and the paracentral areas within the same hemifield in the mild, moderate, and severe groups of CACG patients. PMID- 14522760 TI - Evaluating several sources of variability for standard and SWAP visual fields in glaucoma patients, suspects, and normals. AB - PURPOSE: To quantify factors affecting test-retest variability of threshold measurements over a series of 3 serial visual fields (VF). DESIGN: Prospective comparative observational study. PARTICIPANTS: Forty-one normals, 10 suspects and 35 stable glaucoma patients. METHODS: All subjects performed 3 standard and 3 short-wavelength automated perimetry (SWAP) VFs. At each VF location, severity (defined as age-corrected total deviation) and test-retest variability (TRV), defined as the standard deviation of 3 serial threshold values, were calculated. A multiple regression model (constructed separately for standard VF and SWAP) incorporated 13 factors: severity, location, eccentricity, study group, diagnosis, superior versus inferior hemifield, nasal versus temporal hemifield, one-versus-two thresholds, age, mean pupil size, pupil size variability, between subject variation, and residual variation. MAIN OUTCOME MEASURES: Variability in threshold sensitivity VF values. RESULTS: Mean TRV (+/- standard deviation) for normal, suspect and glaucoma eyes, respectively, was: 1.28 +/- 0.87, 1.53 +/- 1.04 and 2.20 +/- 1.79 dB for standard VF, and 1.87 +/- 1.35, 1.86 +/- 1.24 and 2.68 +/- 1.85 dB for SWAP. The contribution of each factor to the model for standard VF and SWAP (SWAP in parentheses) were: severity 15.5% (6.9%); location 2.7% (4.1%); eccentricity 1.1% (0.64%); diagnosis 2.9% (5.9%); "superior versus inferior" hemifield 0.17% (1.7%); "nasal versus temporal" hemifield 0.06% (0.02%); one-versus-two thresholds 0.04% (0.16%); age 0.1% (0.06%); mean pupil size 0.59% (0.1%); pupil size variability 3.2% (2.8%); between-subject 8.0% (13.5%) and residual variation 61.0% (66.6%). Excluding between-subject and residual variation, the 11-factor model was able to account for less than one third of the variability seen in both standard VF and SWAP. CONCLUSIONS: Severity of defect and between subject variation exerted the largest effect on TRV. However, even if all 11 factors could be adjusted for, it would reduce the magnitude of TRV by only 30%. More work is needed to reduce the remaining variability inherent in psychophysical testing and to better understand the intrinsic physiological variability present both in healthy and diseased eyes. It is possible that a larger number of VFs used for the calculation of TRV might further reduce the magnitude of the remaining variability found in this study. PMID- 14522761 TI - Corneal thickness measurements and frequency doubling technology perimetry abnormalities in ocular hypertensive eyes. AB - OBJECTIVE: It has been suggested that some patients currently diagnosed with ocular hypertension (OHT) may have thicker than average corneas that result in an overestimation of their true intraocular pressure. Consequently, patients with OHT with greater corneal thickness may be at a lower risk for functional glaucomatous damage, including visual field loss measured with frequency doubling technology (FDT) perimetry. The purpose of this study was to evaluate the frequency of FDT perimetry deficits in patients with OHT and to correlate these findings with central corneal thickness (CCT) measurements. DESIGN: Observational case control study. PARTICIPANTS AND CONTROLS: Sixty-five patients with OHT with normal optic discs and normal standard achromatic automated perimetry (SAP) visual fields and 52 normal control subjects. METHODS: All participants underwent SAP, FDT perimetry, and CCT measurements using ultrasound pachymetry. MAIN OUTCOME MEASURES: CCT measurements in patients with OHT with abnormal repeatable FDT test results were compared with CCT measurements in patients with OHT with normal FDT results. In addition, CCT measurements in patients with OHT were compared with CCT measurements in normal control subjects. RESULTS: Fourteen of 65 patients with OHT (21.5%) demonstrated repeatable FDT abnormalities. The mean CCT of patients with OHT with abnormal FDT results was significantly lower than the mean CCT of patients with OHT with normal FDT results (542+/-35 microm versus 575+/-35 microm; P = 0.003). The mean CCT in the normal control group was 556+/ 36 microm. The mean CCT in patients with OHT with normal FDT results was significantly higher than in normal subjects (P = 0.008). No statistically significant difference was found between mean CCT in normal subjects and in patients with OHT with abnormal FDT results (P = 0.18). CONCLUSIONS: Patients currently diagnosed with OHT, but with visual field loss detected by FDT perimetry, had significantly lower CCT measurements than patients with OHT with normal FDT results. These findings suggest that patients with OHT with thinner corneas are more likely to develop early glaucomatous functional damage and that CCT measurements should be taken into account when assessing risk for the development of glaucoma among OHT subjects. PMID- 14522762 TI - Outcome of surgery on infants younger than 1 month with congenital glaucoma. AB - PURPOSE: To determine the visual outcomes and surgical and anesthetic complication rates of patients with newborn glaucoma operated within 1 month of age. DESIGN: Retrospective, consecutive, noncomparative case series. PARTICIPANTS: All children with newborn glaucoma who underwent surgery between January 1990 and December 2000 were included. METHODS: The medical records of 25 consecutive patients (47 eyes) who underwent primary combined trabeculotomy and trabeculectomy either bilaterally in a single session or unilaterally were reviewed retrospectively. Outcomes were evaluated using Kaplan-Meier survival analysis. MAIN OUTCOME MEASURES: Clinical outcome assessment included corneal clarity, intraocular pressure (IOP), bleb characteristics, visual acuity, refractive errors, and identification of surgical and anesthetic complications. RESULTS: The mean follow-up was 3.1+/-1.8 years (range, 9.5 months-7.4 years). The mean preoperative IOP was 26.9+/-5.2 mmHg (range, 14-42 mmHg). At the final follow-up visit, the mean IOP was 14.5+/-3.8 mmHg (range, 8-28 mmHg). The percentage reduction in IOP was 43.3+/-21.5 (P<0.0001). Twelve-, 24-, and 36 month survival rates for complete success for IOP control were 89.4%, 83.6%, and 71.7%, respectively, which were maintained for 7 years of follow-up. After surgery, complete clearance of corneal edema was achieved in 66% of the eyes. Data on visual acuity was available for 19 patients. Final best spectacle corrected visual acuity was 20/40 or better in the better eye in 5 patients (26.3%), 7 patients (36.8%) obtained 20/60 or better in the better eye, 8 patients (42.1%) achieved final visual acuity of less than 20/60 to 20/200 in the better eye (low vision), and four patients obtained less than 20/400 visual acuity in the better eye (blind) according to World Health Organization criteria. However, there was no eye with absent perception of light in the better eye. Myopia (mean spherical equivalent, 4.6+/-3.2 diopters) was the most common refractive error, present in approximately half of the eyes (n = 23; 53.8%). There were no significant intraoperative or postoperative complications in any patient. Anesthesia-related complications developed in 2 patients; however, they were resuscitated successfully. CONCLUSIONS: Primary combined trabeculotomy trabeculectomy offers a viable surgical option in infants that have cloudy corneas at birth as a result of congenital glaucoma. It is associated with a favorable visual outcome and a low rate of anesthetic complications in an Indian population. PMID- 14522763 TI - Effect of oral acyclovir after penetrating keratoplasty for herpetic keratitis: a placebo-controlled multicenter trial. AB - OBJECTIVE: To determine the prophylactic effect of oral acyclovir on the recurrence rate of herpetic eye disease after penetrating keratoplasty. DESIGN: A randomized, double-masked, placebo-controlled multicenter trial. PARTICIPANTS: Sixty-eight consecutive patients (68 eyes) with corneal opacities due to herpetic eye disease who underwent penetrating keratoplasty. INTERVENTION: Oral acyclovir 400 mg twice daily or placebo tablets for 6 months. MAIN OUTCOME MEASURES: The recurrence rate of herpetic eye disease-related events and rejection episodes, proven by viral cell culture or polymerase chain reaction. RESULTS: During the 2 year follow-up period, there were 3 culture-proven herpetic eye disease recurrences in the acyclovir group and 9 in the placebo group. Lifetime survival analysis of the probability of remaining free from recurrence revealed a significantly reduced risk of recurrent herpetic disease in the acyclovir-treated group. CONCLUSION: This study suggests that oral acyclovir effectively prevents herpes-related recurrences after penetrating keratoplasty in herpetic eye disease. PMID- 14522765 TI - Recurrence of keratoconus characteristics: a clinical and histologic follow-up analysis of donor grafts. AB - PURPOSE: To report on clinical corneal topography, histopathologic analysis, and fine structure findings in failed grafts after penetrating keratoplasty (PK) for keratoconus (KC). DESIGN: Retrospective, consecutive, interventional case series with histologic and clinical correlation. PARTICIPANTS: Twelve corneal buttons were obtained from consecutive patients undergoing repeated PK 10 to 28 years after the initial PK for KC. The indication for regrafting was endothelial deficiency in seven cases, irreversible immune graft rejection in two cases, and corneal ectasia in three cases. METHODS: Removed corneal buttons were examined by light and transmission electron microscopy. A potential correlation between the clinical and videokeratoscopic findings and the microscopic structural observations was analyzed. RESULTS: Preoperative simulated keratometry measured by TMS-1 (Tomey, New York, NY) or EyeSys CAS (EyeSys Technology, Houston, TX) ranged from 49.8 to 66.1 diopters. A pattern compatible with KC characteristics was observed in all cases. Fine structure analysis revealed Bowman's layer disruption or folds and stromal deposits in all corneal buttons. However, central corneal thinning was not present in any of the removed buttons. CONCLUSIONS: Structure changes compatible with the diagnosis of KC were observed in all donor buttons many years after PK on KC recipients. Recurrence of the KC characteristics may result from graft repopulation by recipients' keratocytes, aging of the grafted tissue, or both. PMID- 14522766 TI - Difference map or single elevation map in the evaluation of corneal forward shift after LASIK. AB - PURPOSE: Forward shift of the cornea after excimer laser refractive surgery has been assessed on a difference map generated from two elevation maps of the scanning-slit corneal topography. The current study was conducted to test whether similar evaluation is possible on a postoperative color-coded elevation map alone. DESIGN: Prospective, noncomparative case series. PARTICIPANTS: One hundred sixty-three eyes of 86 patients with myopic refractive errors of -1 to -13.50 diopters. INTERVENTION: LASIK was performed. Corneal topography of the posterior corneal surface was obtained with the scanning-slit topography system before and 1 month after surgery. MAIN OUTCOME MEASURES: The amount of forward shift of the posterior corneal surface was determined at the center of the difference map generated from preoperative and postoperative elevation maps. For surface alignment in the difference map, the 3-mm wide peripheral annular fit-zone was used. The eyes were classified into two groups depending on the amount of forward shift, using 50 micro m as the threshold. Next, on the single postoperative color coded elevation map, which is drawn relative to the individual best-fit sphere, the eye was judged to be abnormal (with significant forward shift) when more than three colors (discriminant number) were found within the central 3-mm area, and sensitivity and specificity were calculated. By varying the discriminant number from 3 to 9, receiver operator characteristic (ROC) curves were created. RESULTS: The ROC curve analyses demonstrated that sufficient true positive ratio (sensitivity) and false-positive ratio (100-specificity [%]) could not be obtained with any discriminant color number when judgments were made on a single color-coded map. There was a weak, but significant, correlation between the amount of corneal forward shift and the radius of curvature of the posterior best fit sphere (Pearson r = -0.170; P = 0.030), indicating that a cornea with greater forward shift tended to be drawn on a steeper best-fit sphere, and thus the forward protrusion of the posterior surface failed to be depicted. CONCLUSIONS: Forward shift of the cornea after excimer laser surgery should be evaluated on the difference map generated from two elevation maps, such as preoperative and postoperative maps. PMID- 14522767 TI - Comparison of flap thickness reproducibility using microkeratomes with a second motor for advancement. AB - PURPOSE: To compare flap thickness and reproducibility of four different types of microkeratomes during LASIK. DESIGN: Retrospective, nonrandomized, comparative case series. PARTICIPANTS: Four hundred ninety consecutive eyes underwent LASIK and were evaluated by measuring the central flap thickness by subtractive pachymetry. METHODS: All flaps were created using the Bausch & Lomb (Miami, FL) Hansatome 180 head, the Alcon (Fort Worth, TX) Summit Krumeich Barraquer Microkeratome 160 head, the Moria (Antony, France) Carriazo Barraquer (CB) 130 head, or the Moria M2 110 head. RESULTS: The flap thickness measurements differed according to the microkeratome used and were 131+/-28 microm in 41 eyes (8.4%) with the Bausch & Lomb Hansatome 180 head, 162+/-21 microm in 127 eyes (25.9%) with the Alcon Summit Krumeich Barraquer Microkeratome (SKBM) 160 head, 157+/-40 microm in 65 eyes (13.3%) with the Moria CB 130 head, and 134+/-23 microm in 257 eyes (52.4%) with the Moria M2 110 head. The central flap thickness with the SKBM and Moria M2 was statistically significantly more reproducible than with the Moria CB (P< 0.0005). There is no correlation between flap thickness reproducibility and age, corneal thickness, or corneal keratometric values. However, considering all the microkeratomes, female gender had statistically significantly more variability than male gender (P<0.02). CONCLUSIONS: Based on these results, the greatest predictability of flap thickness was seen with the SKBM and Moria M2 microkeratomes, which both use a second motor for advancement. The greatest variability, noted with the Moria CB, was likely due to the manual translation feature and places further importance on the safety of the second motor and automation when performing LASIK. PMID- 14522768 TI - Lens epithelial cell reaction after implantation of different intraocular lens materials: two-year results of a randomized prospective trial. AB - PURPOSE: To determine the influence of intraocular lens (IOL) material on anterior capsular opacification and membrane growth over the anterior IOL surface in patients who have undergone standardized small-incision cataract surgery and foldable IOL implantation in the capsular bag. DESIGN: Randomized controlled trial. PARTICIPANTS: Eighty-eight cataract patients (88 eyes). METHODS: Patients were randomly assigned to receive one of four different foldable IOLs after phacoemulsification: Storz Hydroview H60M, Corneal ACR6D, AMO SI40NB, and Alcon AcrySof MA60BM. Examinations on days 7, 30, 90, 180, 360, and 720 after surgery included ophthalmologic examination, slit-lamp biomicroscopy, and photography using red reflex and focal illumination of the anterior IOL surface. MAIN OUTCOME MEASURES: Best-corrected visual acuity was measured at each examination. In addition, the anterior capsule opacification and the membrane growth on the anterior IOL surface were graded according to a subjective method by the same researcher. RESULTS: The fibrosis of the anterior capsule was more frequently observed in the group using Corneal ACR6D and AMO SI40NB. The Hydroview and ACR6D groups showed a higher percentage of cases with membrane growth from the rhexis edge on the anterior IOL surface. AcrySof showed the lowest presence of fibrosis of the anterior capsule, and no membrane growth was noted. CONCLUSIONS: Anterior capsule opacification is an index of IOL biocompatibility. The natural location of lens epithelial cells (LECs) precludes the possibility of the IOL's design influencing the anterior capsule behavior. The local response of LECs varies according to the IOL studied. This may be related to the chemical and physical properties of the materials used in the different IOLs. PMID- 14522769 TI - Phacoanaphylaxis in Behcet's disease: a clinicopathologic and immunohistochemical study. AB - PURPOSE: To describe the occurrence of phacoanaphylaxis in enucleated eyes of patients with Behcet's disease. DESIGN: Retrospective, observational, case series and human tissue study. PARTICIPANTS: Twenty-six patients with Behcet's disease who underwent enucleation. METHODS: Histopathologic analysis was performed on 28 enucleated eyes of 26 patients with Behcet's disease. The eyes were divided into two groups, based on the absence or presence of tractional retinal detachment associated with cyclitic membrane formation. Selected eyes with tractional retinal detachment were stained for immunohistochemical examination. MAIN OUTCOME MEASURES: Histopathologic examination of enucleated eyes, including routine histopathologic and immunohistochemical studies. RESULTS: None of the five eyes without retinal detachment showed phacoanaphylaxis. Nine of the 23 eyes with detachment exhibited phacoanaphylaxis, 10 showed no inflammation of the lens, and four were aphakic. There was marked inflammatory cell infiltration in the cyclitic membrane of all nine eyes with phacoanaphylaxis. Immunohistochemical examination demonstrated positive staining for the macrophage markers in the epithelioid and giant cells. The average interval between onset of the ocular manifestations of Behcet's disease and enucleation was 63 months for eyes with phacoanaphylaxis and 35 months for eyes without phacoanaphylaxis (P<0.005). CONCLUSIONS: In Behcet's disease, eyes with long-standing intraocular inflammation complicated by cyclitic membrane formation may develop phacoanaphylaxis. Such patients may benefit from surgical removal of the cyclitic membrane along with the lens in eyes with significant visual function. PMID- 14522770 TI - Antibiotic resistance patterns of ocular bacterial flora: a prospective study of patients undergoing anterior segment surgery. AB - PURPOSE: To determine the antibiotic susceptibility patterns of conjunctival bacterial flora isolated preoperatively from patients undergoing anterior segment surgery. DESIGN: Prospective observational study. PARTICIPANTS: One hundred fifty six eyes from 139 patients scheduled for anterior segment surgery were enrolled over a 6-month period from August 2001 to February 2002. METHODS: Conjunctival cultures were obtained on the day of surgery before povidone-iodine or antibiotic application. MAIN OUTCOME MEASURES: Bacterial isolates were identified and tested for antibiotic susceptibility using the Kirby-Bauer disc-diffusion technique. RESULTS: Among the 156 eyes studied, 36 were from patients who had undergone either bilateral surgery or more than one surgery in the same eye. Only the first eyes of the 120 patients that underwent initial ocular surgery were included in our analysis. Of these 120 eyes, 21 (18%) showed no bacterial growth. Of the 143 bacterial strains isolated from the remaining 99 eyes, 112 (78%) were coagulase negative staphylococci (CNS). Among the CNS, greater than 90% were susceptible to cefotaxime, levofloxacin, imipenem, meropenem, vancomycin, and each of the aminoglycosides except neomycin. Between 70% and 90% of the CNS were susceptible to cefazolin, neomycin, ciprofloxacin, ofloxacin, norfloxacin, and chloramphenicol. Less than 70% of the isolated CNS were sensitive to the penicillin analogues, ceftazidime, erythromycin, and tetracycline. CONCLUSIONS: Preoperative conjunctival isolates of CNS seem to be most sensitive to vancomycin, the aminoglycosides (except neomycin), and levofloxacin. PMID- 14522771 TI - Longitudinal rates of annual eye examinations of persons with diabetes and chronic eye diseases. AB - OBJECTIVE: To assess the rate of annual eye examinations over time among older Americans with diabetes and chronic eye diseases. DESIGN: Longitudinal analysis of Medicare claims data. PARTICIPANTS: Random sample of Medicare beneficiaries aged 65 years or older. METHODS: Beneficiaries were followed between 1991 and 1999, unless mortality or enrollment in a health maintenance organization for > 6 months in a given 12-month period intervened. All claims data (both physician and facility) during this time were analyzed for the presence of International Classification of Diseases 9 codes consistent with 1 of the 3 study conditions and the performance of eye examinations. MAIN OUTCOME MEASURES: Claims submitted by optometrists, ophthalmologists, or other providers of eye care for subjects with diabetes, glaucoma, or age-related macular degeneration (ARMD). Rates were calculated on the basis of a 15-month time window for annual examinations rather than for 12 months to allow for less than full compliance with the guidelines for various reasons (e.g., bad weather). RESULTS: Among those with diabetes in this population, 50% to 60% had annual eye examinations in a 15-month period. Of those followed for at least 75 months after diagnosis, about three quarters had one or more 15-month gaps between visits. For subjects diagnosed with glaucoma, most visit rates were in the 70% to 90% range per 15-month period. The percentage of subjects with at least one 15-month period with no visits was considerably lower than for diabetes. The patterns for those with ARMD were in between those for diabetes and glaucoma. Over a nine-year period, only slightly over half of persons with at least one of the study conditions complied with practice guidelines. CONCLUSIONS: Annual eye examinations for persons diagnosed with diabetes, glaucoma, and ARMD are important for detecting potentially treatable vision loss among those already diagnosed with these conditions. Currently, actual rates of eye examinations for persons diagnosed with the study conditions fall far short of recommended rates. As such, approaches to enhancing longitudinal follow-up of those already in the eye care system are needed. PMID- 14522772 TI - Cataract surgery and the 5-year incidence of late-stage age-related maculopathy: pooled findings from the Beaver Dam and Blue Mountains eye studies. AB - PURPOSE: To assess whether cataract surgery in older persons increases risk for the development of late-stage age-related maculopathy (ARM). DESIGN: Combined analysis of longitudinal data from two population-based cohorts, the Beaver Dam Eye Study and Blue Mountains Eye Study. PARTICIPANTS: The Beaver Dam Eye Study examined 4926 persons aged 43 years or older at baseline and re-examined 3684 after 5 years. The Blue Mountains Eye Study examined 3654 persons aged 49 years or older at baseline and re-examined 2335 after 5 years. METHODS: The two studies used similar protocols for retinal photography and photographic grading. We defined incident late-stage ARM as the development of neovascular ARM or geographic atrophy in eyes without either lesion type at baseline that was confirmed by consensus between the study investigators. Nonphakic eyes included eyes that were aphakic or pseudophakic at baseline. Eye-specific data were analyzed. Age- and study site-adjusted relative risks were calculated using the Cochran-Mantel-Haenszel method. Multivariate-adjusted odds ratios (ORs) were also estimated using generalized estimating equation models. RESULTS: Of the 6019 participants examined after 5 years, 11,391 eyes were considered at risk for developing late-stage ARM, including 315 nonphakic and 11,076 phakic eyes. Late state ARM (either neovascular ARM or geographic atrophy) developed in 6.0% to 7.5% of nonphakic eyes (10 of 168 right and 11 of 147 left eyes), compared with 0.7% of phakic eyes (40 of 5504 right and 37 of 5572 left eyes) during the 5-year period. Age- and study site-adjusted 5-year relative risks were 2.8 (95% confidence interval [CI], 1.6-5.1) for right and 3.7 (95% CI, 2.1-6.4) for left eyes. After further adjustment for gender, smoking, and the presence of indistinct or reticular drusen or pigmentary abnormalities at baseline, nonphakic eyes had a substantially higher risk for developing either late-stage ARM lesion compared with phakic eyes, OR = 5.7 (95% CI, 2.4-13.6). CONCLUSIONS: Pooled findings from these two large population-based cohorts support the hypothesis that cataract surgery in older persons may be associated with an increased subsequent risk for developing late-stage ARM, particularly neovascular ARM. PMID- 14522773 TI - Solitary congenital hypertrophy of the retinal pigment epithelium: clinical features and frequency of enlargement in 330 patients. AB - OBJECTIVE: To describe the clinical features of solitary congenital hypertrophy of the retinal pigment epithelium (CHRPE) and to determine the frequency of enlargement of this lesion DESIGN: Retrospective, observational, noncomparative case series. PARTICIPANTS: Three hundred thirty consecutive patients with solitary CHRPE. MAIN OUTCOME MEASURES: The 3 main outcome measures included flat lesion enlargement, intralesional lacunae enlargement, and development of an elevated nodule within the lesion. The clinical features at the time of presentation were analyzed for their impact on the main outcomes using a series of Cox proportional hazards regressions. RESULTS: The most common referring diagnosis included choroidal nevus (26%), choroidal melanoma (15%), CHRPE (9%), and unspecified lesion (48%). The median age at diagnosis was 45 years (range, 1 80 years), and there were no patients with familial adenomatous polyposis or related colon cancer, although a history of cancer was noted in 8% of patients, most commonly breast cancer (3%). The lesion most frequently was located inferotemporally (31%) and at the equatorial region (45%). Rarely, it was located in the macula (1%) or peripapillary region (1%). The median largest basal diameter was 4.5 mm, and the lesion was flat in all cases except in 5 (1.5%), in which there was an intralesional lesion nodule. The lesion was pigmented in 88% of cases and nonpigmented in 12%. Lacunae were noted in 43% of the pigmented CHRPE, and the lacunae showed gradual enlargement in 32%. Factors related to lacunae enlargement included number and relative size of lacunae. Flat enlargement of the lesion was documented in 46% of patients with comparative photographic follow-up and in 83% of those followed up for more than 3 years. The median rate of enlargement was 10 micro m per month. The most important factor associated with flat lesion enlargement was relative size of the lacunae within CHRPE. There were no cases of CHRPE in which a nodule developed while the patient was being followed up. Of the 5 lesions that had a nodule, progressive enlargement of the nodule was found in 3. CONCLUSIONS: Congenital hypertrophy of the retinal pigment epithelium generally has been regarded as a benign, stable lesion, but subtle, flat enlargement was noted in most patients (83%) followed up for 3 or more years using meticulous photographic comparison. Flat enlargement of the lesion appeared to be related to percentage of the mass occupied by lacunae. PMID- 14522774 TI - Stereotactic radiotherapy of symptomatic circumscribed choroidal hemangiomas. AB - PURPOSE: To determine feasibility of low-dose stereotactic radiotherapy in the treatment of symptomatic circumscribed choroidal hemangioma. DESIGN: Prospective, noncomparative, interventional case series. PARTICIPANTS: Five consecutive patients with perifoveolar and peripapillary circumscribed choroidal hemangioma and visual symptoms from exudative retinal detachment. METHODS: A dose of 20 Gy was delivered stereotactically with linear accelerator. Tumor dimensions were determined by B-scan ultrasonography. MAIN OUTCOME MEASURES: Resolution of subretinal fluid, best-corrected visual acuity, and reduction in tumor height. RESULTS: Median tumor height at baseline was 2.8 mm (range, 2.0-4.2 mm). Two tumors were subfoveolar, two were juxtafoveolar, and one was extrafoveolar. Cystic macular edema and subretinal fibrosis were present in both eyes with subfoveolar tumor. Exudative retinal detachment resolved within a median of 5 months (response rate, 100%; 95% CI, 48%-100%). Median best-corrected visual acuity was 20/50 (range, 20/22-20/100) at diagnosis and 20/25 (range, 20/20 20/60) 20 months after treatment. Tumor height had decreased a median of 24% (range, 0%-31%) by 6 months and 29% (range, 9%-59%) by 20 months. Secondary retinal pigment epithelial mottling associated with tumor regression occurred in two patients. One eye developed a paracentral scotoma. CONCLUSIONS: Stereotactic radiotherapy can be targeted precisely enough to induce regression of subretinal fluid from circumscribed choroidal hemangiomas. PMID- 14522776 TI - New retinoblastoma tumor formation in children initially treated with systemic carboplatin. AB - PURPOSE: To determine the frequency and timing of new intraocular tumor formation in children with hereditary retinoblastoma initially treated with systemic carboplatin. DESIGN: Retrospective, noncomparative case series. PARTICIPANTS: This study included 34 children (57 eyes) with hereditary bilateral retinoblastoma initially treated with systemic carboplatin at the Robert M. Ellsworth Ophthalmic Oncology Center at NewYork-Presbyterian Hospital from 1994 through 2000. MAIN OUTCOME MEASURES: New tumor formation after initial treatment with systemic carboplatin. RESULTS: There were a total of 165 tumors in 57 eyes. There were 63 new tumors in 27 eyes (47%) after administration of systemic carboplatin, for a mean of 1.1 new tumors per eye. The mean patient age at time of new tumor presentation was 9 months, with 57% of new tumors developing within 4 months of carboplatin treatment. Kaplan-Meier analysis showed that children who were treated when younger than 6 months of age were more likely to have new tumors (60%) compared with those treated after 6 months of age (31%; P = 0.0182). CONCLUSIONS: New intraocular tumors continue to develop after systemic carboplatin; most new tumors appeared within 4 months of treatment. PMID- 14522775 TI - Retinoblastoma associated with chromosomal 13q14 deletion mosaicism. AB - PURPOSE: To assess the risk of retinoblastoma developing in children with microscopic chromosomal with mosaic deletions involving 13q14. DESIGN: Case report and systematic literature review. PARTICIPANTS: Data on 29 patients with a mosaic and 107 patients with a nonmosaic somatic deletion of chromosome 13q14 were compared. MAIN OUTCOME MEASURES: Age at diagnosis, frequency, and laterality of retinoblastoma. CASE REPORT: A dysmorphic baby, who carried a chromosomal deletion involving 13q14 in 34% of peripheral blood lymphocytes, had neuroradiologic evidence of retinoblastoma at the age of 2 weeks. She developed trilateral retinoblastoma, a pineal neuroblastic tumor, at the age of 10 months. The diagnosis of her tumor was delayed because of misjudgment of risk of retinoblastoma developing. RESULTS: Meta-analysis revealed no difference between children with mosaic and nonmosaic chromosomal deletion of 13q14 regarding the age at diagnosis, laterality of tumor, and presence of family history for retinoblastoma. A lower percentage of somatic cells with mosaic deletion did not predict a higher age at diagnosis or unilateral tumors. No statistically significant difference was noted regarding the presence of mental retardation, dysmorphic features, and anomalies of internal organs between mosaic and nonmosaic deletions. Only 7% (95% confidence interval, 1-23) of 29 patients who had a mosaic chromosomal deletion including 13q14 were not reported to develop retinoblastoma. CONCLUSIONS: Whenever a 13q14 deletion is diagnosed, immediate ophthalmologic evaluation is recommended to ensure prompt diagnosis of retinoblastoma. Mosaic and nonmosaic chromosomal deletions of 13q14 do not differ regarding the risk and type of retinoblastoma developing. PMID- 14522778 TI - Osteogenic sarcoma after orbital radiation rhabdomyosarcoma. AB - PURPOSE: We describe the occurrence of maxillary and orbital osteogenic sarcoma in a child after treatment of contralateral orbital rhabdomyosarcoma with external beam radiation and chemotherapy. DESIGN: Interventional case report. INTERVENTION: Treatment of a maxillary and orbital rhabdomyosarcoma with surgical resection, systemic chemotherapy, and local radiation. MAIN OUTCOME MEASURES: Occurrence and histology of secondary malignancy after orbital radiation. RESULTS: An eleven year-old male presented for evaluation of left facial swelling, occurring ten years after successful treatment of a right orbital embryonal rhabdomyosarcoma with chemotherapy and 5040 cGY of external beam radiation. Computerized tomography demonstrated a mass involving the left maxillary sinus and orbital floor. Biopsy demonstrated osteogenic sarcoma. Despite attempted total excision with radical maxillectomy, resection margins were found to have microscopic extension of the tumor. Postoperatively he was treated with systemic chemotherapy and local radiation. Eight months postoperatively he remains alive despite local progression. CONCLUSIONS: Osteogenic sarcoma can occur as a secondary malignancy years after the successful treatment of orbital rhabdomyosarcoma with external beam radiation and chemotherapy. After orbital radiation, subjects should undergo routine lifelong examinations. PMID- 14522779 TI - Orbital and periorbital myofibromas in childhood: two case reports. AB - PURPOSE: Infantile myofibromatosis is an uncommon tumor that occurs rarely in the periorbit and orbit. This article reports two cases of infantile myofibromatosis of the orbital adnexa and describes the associated clinical, histopathologic, and immunohistochemical findings. DESIGN: Two retrospective, interventional case reports with clinicopathologic correlation. INTERVENTION: Treatment consisted of excision of the tumors. MAIN OUTCOME MEASURES: Histologic and immunohistochemical evaluation and clinical evaluation for tumor recurrence. RESULTS: The first patient was a newborn male with a large tumor extending from his eyelid that was excised at day 2 of life. Histologic and immunohistochemistry analyses were used to make a diagnosis of infantile myofibromatosis. He remains disease free at age 7 years. The second case was a 6-year-old boy with a 1-month history of proptosis resulting from an orbital mass. Incisional biopsy revealed a tumor consistent with infantile myofibromatosis. He remains tumor free 12 months after complete gross surgical resection. CONCLUSIONS: Infantile myofibromatosis is an uncommon tumor that is rare in the orbit. Differential diagnosis can be difficult based solely on histologic analysis. Immunohistochemistry evaluation demonstrating cytoplasmic actin filaments within neoplastic spindle cells confirms the diagnosis. As soon as the diagnosis is made, chest and abdominal imaging is of value to define the prognosis and to direct further treatment. After the diagnosis of nonvisceral infantile myofibromatosis, complete gross resection, if possible, is the treatment of choice. PMID- 14522780 TI - Primary cystadenocarcinoma of the lacrimal gland. AB - PURPOSE: To report a patient with a cystadenocarcinoma of the lacrimal gland, a tumor not previously described in the ophthalmic literature. Salivary gland cystadenocarcinomas constitute a distinct group of epithelial malignancies characterized by an invasive, predominantly cystic pattern of growth that have an indolent behavior and a low incidence of metastases and recurrences. DESIGN: Single interventional case report. METHODS: The clinical findings, results of imaging studies, and pathologic findings are presented. RESULTS: A 67-year-old man presented with a 5-year history of ptosis in the right upper eyelid. A lacrimal fossa tumor was found. The tumor was excised with an intact capsule, and the histopathologic diagnosis was primary cystadenocarcinoma of the lacrimal gland. The patient received no other form of treatment and has been observed for 1 year without evidence of recurrence or metastatic disease. CONCLUSIONS: Until recently, primary adenocarcinomas of the lacrimal gland were not further subclassified. Current knowledge gained from salivary gland tumors indicates that primary adenocarcinoma encompasses a group of tumors with separate morphologic features and varied biologic behavior. PMID- 14522781 TI - Mapped serial excision for periocular lentigo maligna and lentigo maligna melanoma. AB - PURPOSE: To report the early cure rate for periocular lentigo maligna (LM) and LM melanoma (LMM), using modified Mohs surgery with vertically cut paraffin-embedded sections (mapped serial excision [MSE]). A secondary aim was to identify differences in the clinical features and outcomes between periocular LM and LMM and those found elsewhere on the head and neck. DESIGN: Prospective, noncomparative, interventional case series. PARTICIPANTS: One hundred thirty-five patients undergoing 141 MSE procedures. METHODS: A prospective series of 141 MSE procedures for LM and LMM over a 10-year period (1993-2002) in a single-center Mohs surgical unit. MAIN OUTCOME MEASURES: Recurrence, site, size of LM or LMM, invasiveness, prior recurrence, clear margin of excision, size of final defect, and number of levels required for complete excision. RESULTS: One hundred forty one MSE procedures, of which 23% (32/141) were for LMM and 19% (27/141) were for periocular lesions. Location or prior recurrence were not predictive of invasive disease; however, the size distribution of the initial lesion (P = 0.0354) and the final defect after MSE (P = 0.0183) were larger in LMM. Thirty-one percent of LM and 14% of LMM less than 1 mm thick required larger than 5-mm and 1-cm margins, respectively, for complete excision. Mean follow-up of 32 months (range, 1-100 months) revealed 4 recurrences (3%), of which two were periocular (P = 0.188). CONCLUSIONS: Our review is the largest prospective series of MSE for LM and LMM and suggests that it is the treatment of choice in these forms of melanoma. Mapped serial excision offers a high early cure rate in conjunction with tissue conservation, which is of particular relevance in the periocular region. There were no significant differences between periocular LM and LMM and those found elsewhere in the head and neck region. It also appears that the current recommendations of 5-mm margins for in situ melanoma (LM) and 1-cm margins for melanoma less than 1 mm thick are insufficient for complete excision of LM or LMM, emphasizing the importance of margin-controlled excision of these lesions. PMID- 14522782 TI - Optic nerve sheath meningiomas. AB - PURPOSE: To study the natural history and growth of optic nerve sheath meningiomas and evaluate their management outcome. DESIGN: Clinicopathologic retrospective noncomparative case series. METHODS: A retrospective study of 88 patients who were treated between 1976 and 1999 at the University of British Columbia and the University of Amsterdam. Clinical reports, imaging studies, and histopathologic findings were reviewed. RESULTS: The mean age at onset of symptoms was 40.3 years, and most were seen in middle-aged females. Patients typically presented with visual loss, frequently associated with optic atrophy or papilledema and occasionally optociliary shunt vessels. On imaging, the optic nerve demonstrated segmental or diffuse thickening of the sheath or globular growth. Calcification was seen in 31% of cases and was associated with slower tumor growth. Tumors with posterior components in the orbit had more frequent intracranial involvement. Intracranial extension was more frequent and had a greater growth rate in younger patients. Irregular margins in the orbit implied local invasion. A presenting visual acuity better than 20/50 correlated with longer preservation of vision. Patients who underwent radiotherapy showed improvement in their visual acuity, and tumor growth was halted. Optic sheath decompression did not preserve vision. En bloc tumor excision was associated with no detectable recurrence in contrast to debulked tumors that recurred. CONCLUSIONS: Meningiomas show characteristic indolent growth. Management therefore should be conservative in most cases. Radiotherapy is indicated in patients with progressive visual deterioration. Surgery, when indicated, should be an en bloc excision. PMID- 14522783 TI - The morphology of an infarct in nonarteritic anterior ischemic optic neuropathy. AB - OBJECTIVE: The mechanism by which nonarteritic anterior ischemic optic neuropathy (NAION) causes an infarct in the optic nerve is controversial. We studied the three-dimensional anatomic configuration of a NAION infarct to better elucidate its pathophysiology. DESIGN: Case report with clinicopathologic correlation. METHODS: Serial sections of the optic nerve from a previously reported patient diagnosed with NAION 20 days before death were studied. Every fourth slide was stained with hematoxylin-eosin, photographed, and digitized. NIH Image 1.62 was used to reconstruct the nerve in all three dimensions, and the infarct morphology was analyzed. MAIN OUTCOME MEASURES: Morphology of the reconstructed optic nerve infarct. RESULTS: The area of axonal loss within each section of the optic nerve was identified and reconstructed. The loss was in the superior part of the nerve, encircling the central retinal artery at its greatest extent. Remaining areas of the nerve appeared healthy, and, notably, the periphery of the uninvolved inferior portion of the nerve was normal. Three-dimensional analysis revealed two distinct areas of infarct at the posterior extent of the lesion which coalesced toward the center of the lesion and finally tapered as the infarct reached the optic nerve head. Sagittal reconstructions gave the appearance of a two-pronged fork posteriorly connecting to a single "handle" anteriorly. There was no obvious correlation between the configuration of the infarct and any single vascular territory. The total length of the nerve involved by the infarct was approximately 1.5 mm. CONCLUSIONS: The morphology of this NAION infarct is not consistent with disease of large or small vessels and, more likely, represents a form of compartment syndrome that causes tissue ischemia. PMID- 14522784 TI - Blindness associated with enlarging mycotic aneurysm after cavernous sinus thrombosis. AB - PURPOSE: We report a patient with an enlarging internal carotid mycotic aneurysm secondary to septic cavernous sinus thrombosis presenting with acute visual loss. DESIGN: Single observational case report. METHODS: Retrospective review of the medical record and review of the literature. RESULTS: A 19-year-old man with residual left sixth nerve palsy and decreased vision in his left eye caused by left cavernous sinus thrombosis secondary to pansinusitis was seen 2 weeks after discharge with acute decreased visual acuity in the right eye. A workup revealed an enlarging left carotid/ophthalmic aneurysm that compressed the optic chiasm and right optic nerve. The patient was taken to the interventional angiography suite, where his left internal carotid artery was occluded endovascularly. The patient's vision improved on discharge. CONCLUSIONS: Visual loss caused by a mycotic carotid aneurysm is an infrequent sequelae after cavernous sinus thrombosis and is not well described in the literature. To our knowledge, this is the first reported case of acute visual loss associated with a mycotic ophthalmic aneurysm. The result of treatment was good in this case, with the patient's visual acuity returning to pretreatment status. PMID- 14522785 TI - Ocular manifestations in Blau syndrome associated with a CARD15/Nod2 mutation. AB - PURPOSE: To report cases of Blau syndrome with a CARD15/Nod2 mutation. DESIGN: Observational and interventional case report. PARTICIPANTS: A 10-year-old Japanese boy (proband) was seen with secondary angle-closure glaucoma (iris bombe), uveitis, skin rashes, and camptodactyly. His sister had posterior synechia and camptodactyly. She had iritis in both eyes during the follow-up period. Both eyes of the father were phthisical because of granulomatous uveitis and secondary glaucoma. The father also had camptodactyly. METHODS: Surgery was performed to release the iris bombe. Ocular inflammation was treated by topical and systemic steroids. Biopsy specimens from the skin rash and from the iris (from iridectomy) were obtained from the proband. Genetic analyses were performed on the proband, his sister, and their mother for a CARD15/Nod2 mutation. MAIN OUTCOME MEASURES: Clinical features, pathologic findings of the skin and iris specimens, and genetic analysis of the CARD15/Nod2 gene. RESULTS: Phacoemulsification, intraocular lens implantation, and peripheral iridectomy released the iris bombe. The biopsy specimen from the skin rash showed noncaseating, granulomatous infiltration with epithelioid cells and lymphocytes. The iridectomy specimen showed nonspecific inflammation. Systemic and topical steroid therapy partly reduced the ocular inflammation. Genetic analyses showed that the proband and his sister had an R334W mutation in the CARD15/Nod2 gene, but their mother was of the wild type. CONCLUSIONS: Blau syndrome should be considered in the differential diagnosis of childhood uveitis. Genetic analysis of the CARD15/Nod2 gene is helpful in the diagnosis. PMID- 14522786 TI - Detection by impression cytologic analysis of conjunctival intraepithelial invasion from eyelid sebaceous cell carcinoma. AB - OBJECTIVE: To demonstrate that conjunctival impression cytologic analysis can detect conjunctival intraepithelial invasion from sebaceous cell carcinoma of the eyelid. DESIGN: Observational case series with cytopathologic correlation. PARTICIPANTS: Four patients with unilateral blepharoconjunctivitis and biopsy proven sebaceous cell carcinoma. METHODS: Impression cytologic analysis specimens were taken from the suspicious area of the bulbar conjunctiva of each patient. Staining of the specimens was performed with a modified Papanicolaou stain. MAIN OUTCOME MEASURE: Observation of the abnormal tumor cells in the collected specimens by bright field microscope. RESULTS: The technique of impression cytologic analysis allowed collection and identification of abnormal tumor cells with characteristic cytoplasmic vacuoles. CONCLUSIONS: Conjunctival impression cytologic analysis successfully detected the ocular surface sebaceous carcinoma cells from the eyelid. However, full-thickness biopsies are necessary to confirm the diagnosis. Judicious use of impression cytologic analysis may facilitate the detection and diagnosis of this invasive tumor. PMID- 14522787 TI - Histoplasma granuloma of the conjunctiva. AB - PURPOSE: To describe the clinical and pathologic features of a conjunctival granuloma resulting from Histoplasma capsulatum. DESIGN: Clinicopathologic case report. METHODS: Surgical excision of conjunctival lesion, histopathologic study with special stains, and a fluorescent antibody technique. RESULTS: A conjunctiva granuloma was found to contain microorganisms, compatible with H. capsulatum, confirmed by a fluorescent antibody technique. The conjunctiva returned to normal. CONCLUSIONS: A focal granuloma of the conjunctiva contained H. capsulatum in a patient whose recent symptoms suggested primary infection. The conjunctival lesion was similar to mucocutaneous lesions seen in patients with systemic histoplasmosis. PMID- 14522788 TI - Orbital implants in enucleation surgery: a report by the American Academy of Ophthalmology. AB - OBJECTIVE: To compare prosthetic and implant motility and the incidence of complications associated with porous and nonporous enucleation implants. METHODS: Literature searches conducted in January 2002 for 1985 to 2001 and May 2002 for October 2001 to 2002 retrieved relevant citations. The searches were conducted in MEDLINE and limited to articles published in English with abstracts. Panel members reviewed the articles for relevance to the assessment questions, and those considered relevant were rated according to the strength of the evidence. RESULTS: A randomized clinical trial and a longitudinal cohort study detected no difference in implant or prosthetic movement between nonpegged hydroxyapatite porous and spherical alloplastic nonporous implants. No controlled studies were retrieved that investigated whether pegging porous implants improves prosthetic movement. Several case series indicate that patients with pegged hydroxyapatite implants have some degree of improved prosthetic motility. Longitudinal cohort studies show that sclera-covered hydroxyapatite implants have higher exposure rates than sclera-covered silicone implants, and unwrapped porous polyethylene implants have higher exposure rates than unwrapped acrylic implants. There are numerous case series that document a wide range of implant exposure rates in patients with various enucleation implants. It is difficult to compare complication rates among implant types because patient populations vary, surgical techniques differ, and follow-up periods are often limited. CONCLUSIONS: Based on one randomized clinical trial, spherical alloplastic nonporous and nonpegged porous enucleation implants provide similar implant and prosthetic motility when they are implanted using similar surgical techniques. Coupling the prosthesis to a porous implant with a motility peg or post appears to improve prosthetic motility, but there are few available data in the literature that document the degree of the improvement. There is a widely variable incidence of porous implant exposure, but certain surgical techniques and the type of wrapping material seem to reduce the exposure rate. Additional research is needed to document the long term incidence of complications related to porous enucleation implants and associated surgical techniques. This includes the use of wrapping materials and what procedural modifications, both surgical and prosthetic, are most effective in reducing these complications. PMID- 14522789 TI - Progression of glaucoma. PMID- 14522791 TI - Interface fluid after LASIK. PMID- 14522793 TI - Artisan lens implantation. PMID- 14522794 TI - Interface fluid after LASIK. PMID- 14522796 TI - Chlamydia pneumoniae and NAION. PMID- 14522798 TI - Error tolerance in Helmholtzian accommodation. PMID- 14522799 TI - ICG and time to growth of tumors. PMID- 14522803 TI - Balloon catheter dilatation. PMID- 14522804 TI - Hyperhomocysteinemia and BRVO. PMID- 14522807 TI - Limbal stem cell deficiency. PMID- 14522808 TI - Ventilatory assist driven by patient demand. PMID- 14522809 TI - Plunging into the chaos of the cytokine/chemokine cocktail in pulmonary fibrosis: how many and how important are they? PMID- 14522811 TI - Acquired neuromuscular disorders in the intensive care unit. PMID- 14522812 TI - Acute severe asthma. AB - Acute severe asthma remains a major economic and health burden. The natural history of acute decompensations is one of resolution and only about 0.4% of patients succumb overall. Mortality in medical intensive care units is higher but is less than 3% of hospital admissions. "Near-fatal" episodes may be more frequent, but precise figures are lacking. However, about 30% of medical intensive care unit admissions require intubation and mechanical ventilation with mortality of 8%. Morbidity and mortality increase with socioeconomic deprivation and ethnicity. Seventy to 80% of patients in emergency departments clear within 2 hours with standardized care. The relapse rate varies between 7 and 15%, depending on how aggressively the patient is treated. The airway obstruction in the 20-30% of people resistant to adrenergic agonists in the emergency department slowly reverses over 36-48 hours but requires intense treatment to do so. Current therapeutic options for this group consist of ipratropium and corticosteroids in combination with beta2 selective drugs. Even so, such regimens are not optimal and better approaches are needed. The long-term prognosis after a near-fatal episode is poor and mortality may approach 10%. PMID- 14522813 TI - American Thoracic Society/European Respiratory Society statement: standards for the diagnosis and management of individuals with alpha-1 antitrypsin deficiency. PMID- 14522814 TI - Renewed plea to authors: avoid abbreviations. PMID- 14522815 TI - Dietary cholesterol alters Na+/K+ selectivity at intracellular Na+/K+ pump sites in cardiac myocytes. AB - A modest diet-induced increase in serum cholesterol in rabbits increases the sensitivity of the sarcolemmal Na+/K+ pump to intracellular Na+, whereas a large increase in cholesterol levels decreases the sensitivity to Na+. To examine the mechanisms, we isolated cardiac myocytes from controls and from rabbits with diet induced increases in serum cholesterol. The myocytes were voltage clamped with the use of patch pipettes that contained osmotically balanced solutions with Na+ in a concentration of 10 mM and K+ in concentrations ([K+]pip) ranging from 0 to 140 mM. There was no effect of dietary cholesterol on electrogenic Na+/K+ current (Ip) when pipette solutions were K+ free. A modest increase in serum cholesterol caused a [K+]pip-dependent increase in Ip, whereas a large increase caused a [K+]pip-dependent decrease in Ip. Modeling suggested that pump stimulation with a modest increase in serum cholesterol can be explained by a decrease in the microscopic association constant KK describing the backward reaction E1 + 2K+ --> E2(K+)2, whereas pump inhibition with a large increase in serum cholesterol can be explained by an increase in KK. Because hypercholesterolemia upregulates angiotensin II receptors and because angiotensin II regulates the Na+/K+ pump in cardiac myocytes in a [K+]pip-dependent manner, we blocked angiotensin synthesis or angiotensin II receptors in vivo in cholesterol-fed rabbits. This abolished cholesterol-induced pump inhibition. Because the epsilon-isoform of protein kinase C (epsilonPKC) mediates effects of angiotensin II on the pump, we included specific epsilonPKC-blocking peptide in patch pipette filling solutions. The peptide reversed cholesterol-induced pump inhibition. PMID- 14522816 TI - Sphingomyelinase activates GLUT4 translocation via a cholesterol-dependent mechanism. AB - A basis for the insulin mimetic effect of sphingomyelinase on glucose transporter isoform GLUT4 translocation remains unclear. Because sphingomyelin serves as a major determinant of plasma membrane cholesterol and a relationship between plasma membrane cholesterol and GLUT4 levels has recently become apparent, we assessed whether GLUT4 translocation induced by sphingomyelinase resulted from changes in membrane cholesterol content. Exposure of 3T3-L1 adipocytes to sphingomyelinase resulted in a time-dependent loss of sphingomyelin from the plasma membrane and a concomitant time-dependent accumulation of plasma membrane GLUT4. Degradation products of sphingomyelin did not mimic this stimulatory action. Plasma membrane cholesterol amount was diminished in cells exposed to sphingomyelinase. Restoration of membrane cholesterol blocked the stimulatory effect of sphingomyelinase. Increasing concentrations of methyl-beta cyclodextrin, which resulted in a dose-dependent reversible decrease in membrane cholesterol, led to a dose-dependent reversible increase in GLUT4 incorporation into the plasma membrane. Although increased plasma membrane GLUT4 content by cholesterol extraction with concentrations of methyl-beta-cyclodextrin above 5 mM most likely reflected decreased GLUT4 endocytosis, translocation stimulated by sphingomyelinase or concentrations of methyl-beta-cyclodextrin below 2.5 mM occurred without any visible changes in the endocytic retrieval of GLUT4. Furthermore, moderate loss of cholesterol induced by sphingomyelinase or low concentrations of methyl-beta-cyclodextrin did not alter membrane integrity or increase the abundance of other plasma membrane proteins such as the GLUT1 glucose transporter or the transferrin receptor. Regulation of GLUT4 translocation by moderate cholesterol loss did not involve known insulin signaling proteins. These data reveal that sphingomyelinase enhances GLUT4 exocytosis via a novel cholesterol-dependent mechanism. PMID- 14522817 TI - Contractile function, sarcolemma integrity, and the loss of dystrophin after skeletal muscle eccentric contraction-induced injury. AB - The purpose of this study was to evaluate the integrity of the muscle membrane and its associated cytoskeleton after a contraction-induced injury. A single eccentric contraction was performed in vivo on the tibialis anterior (TA) of male Sprague-Dawley rats at 900 degrees /s throughout a 90 degrees -arc of motion. Maximal tetanic tension (Po) of the TAs was assessed immediately and at 3, 7, and 21 days after the injury. To evaluate sarcolemmal integrity, we used an Evans blue dye (EBD) assay, and to assess structural changes, we used immunofluorescent labeling with antibodies against contractile (myosin, actin), cytoskeletal (alpha actinin, desmin, dystrophin, beta-spectrin), integral membrane (alpha- and beta dystroglycan, sarcoglycan), and extracellular (laminin, fibronectin) proteins. Immediately after injury, P0 was significantly reduced to 4.23 +/- 0.22 N, compared with 8.24 +/- 1.34 N in noninjured controls, and EBD was detected intracellularly in 54 +/- 22% of fibers from the injured TA, compared with 0% in noninjured controls. We found a significant association between EBD-positive fibers and the loss of complete dystrophin labeling. The loss of dystrophin was notable because organization of other components of the subsarcolemmal cytoskeleton was affected minimally (beta-spectrin) or not at all (alpha- and beta-dystroglycan). Labeling with specific antibodies indicated that dystrophin's COOH terminus was selectively more affected than its rod domain. Twenty-one days after injury, contractile properties were normal, fibers did not contain EBD, and dystrophin organization and protein level returned to normal. These data indicate the selective vulnerability of dystrophin after a single eccentric contraction induced injury and suggest a critical role of dystrophin in force transduction. PMID- 14522818 TI - Effects of p38MAPK isoforms on renal mesangial cell inducible nitric oxide synthase expression. AB - Several related isoforms of p38MAPK have been identified and cloned in many species. Although they all contain the dual phosphorylation motif TGY, the expression of these isoforms is not ubiquitous. p38alpha and -beta2 are ubiquitously expressed, whereas p38gamma and -delta appear to have more restricted expression. Because there is evidence for selective activation by upstream kinases and selective preference for downstream substrates, the functions of these conserved proteins is still incompletely understood. We have demonstrated that the renal mesangial cell expresses the mRNA for all the isoforms of p38MAPK, with p38alpha mRNA expressed at the highest level, followed by p38gamma and the lowest levels of expression by p38beta2 and -delta. To determine the functional effects of these proteins on interleukin (IL)-1beta induced inducible nitric oxide synthase (iNOS) expression, we transduced TAT-p38 chimeric proteins into renal mesangial cells and assessed the effects of wild type and mutant p38 isoforms on ligand induced iNOS expression. We show that whereas p38gamma and -delta had minimal effects on iNOS expression, p38alpha and beta2 significantly altered its expression. p38alpha mutant and p38beta2 wild type dose dependently inhibited IL-1beta-induced iNOS expression. These data suggest that p38alpha and beta2 have reciprocal effects on iNOS expression in the mesangial cell, and these observations may have important consequences for the development of selective inhibitors targeting the p38MAPK family of proteins. PMID- 14522819 TI - Mitochondrial coupling in vivo in mouse skeletal muscle. AB - The coupling of mitochondrial ATP synthesis and oxygen consumption (ratio of ATP and oxygen fluxes, P/O) plays a central role in cellular bioenergetics. Reduced P/O values are associated with mitochondrial pathologies that can lead to reduced capacity for ATP synthesis and tissue degeneration. Previous work found a wide range of values for P/O in normal mitochondria. To measure mitochondrial coupling under physiological conditions, we have developed a procedure for determining the P/O of skeletal muscle in vivo. This technique measures ATPase and oxygen consumption rates during ischemia with 31P magnetic resonance and optical spectroscopy, respectively. This novel approach allows the independent quantitative measurement of ATPase and oxygen flux rates in intact tissue. The quantitative measurement of oxygen consumption is made possible by our ability to independently measure the saturations of hemoglobin (Hb) and myoglobin (Mb) from optical spectra. Our results indicate that the P/O in skeletal muscle of the mouse hindlimb measured in vivo is 2.16 +/- 0.24. The theoretical P/O for resting muscle is 2.33. Systemic treatment with 2,4-dinitrophenol to partially uncouple mitochondria does not affect the ATPase rate in the mouse hindlimb but nearly doubles the rate of oxygen consumption, reducing in vivo P/O to 1.37 +/- 0.22. These results indicate that only a small fraction of the oxygen consumption in resting mouse skeletal muscle is nonphosphorylating under physiological conditions, suggesting that mitochondria are more tightly coupled than previously thought. PMID- 14522820 TI - Activation of calcium release assessed by calcium release-induced inactivation of calcium current in rat cardiac myocytes. AB - In mammalian cardiac myocytes, calcium released into the dyadic space rapidly inactivates calcium current (ICa). We used this Ca2+ release-dependent inactivation (RDI) of ICa as a local probe of sarcoplasmic reticulum Ca2+ release activation. In whole cell patch-clamped rat ventricular myocytes, Ca2+ entry induced by short prepulses from -50 mV to positive voltages caused suppression of peak ICa during a test pulse. The negative correlation between peak ICa suppression and ICa inactivation during the test pulse indicated that RDI evoked by the prepulse affected only calcium channels in those dyads in which calcium release was activated. Ca2+ ions injected during the prepulse and during the subsequent tail current suppressed peak ICa in the test pulse to a different extent. Quantitative analysis indicated that equal Ca2+ charge was 3.5 times less effective in inducing release when entering during the prepulse than when entering during the tail. Tail Ca2+ charge injected by the first voltage dependent calcium channel (DHPR) openings was three times less effective than that injected by DHPR reopenings. These findings suggest that calcium release activation can be profoundly influenced by the recent history of L-type Ca2+ channel activity due to potentiation of ryanodine receptors (RyRs) by previous calcium influx. This conclusion was confirmed at the level of single RyRs in planar lipid bilayers: using flash photolysis of the calcium cage NP-EGTA to generate two sequential calcium stimuli, we showed that RyR activation in response to the second stimulus was four times higher than that in response to the first stimulus. PMID- 14522821 TI - Myocyte contractile activity modulates norepinephrine cytotoxicity and survival effects of neuregulin-1beta. AB - The purpose of this study is to test the hypothesis that mechanical and electrical activity in adult rat ventricular myocytes (ARVM) alters responses to proapoptotic and prosurvival ligands. The effects of electrical stimulation on myocyte survival, stress signaling, response to beta-adrenergic receptor (beta AR)-stimulated apoptosis, and neuregulin-1beta (NRG) were examined. Electrical stimulation (6.6 V/cm; 0, 2, and 5 Hz; 2-ms duration; alternating polarity) of ARVM resulted in more than 70% capture. Although ARVM paced for 48 h showed higher mitochondrial uptake of 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (P < 0.05, 0 vs. 2 and 5 Hz), electrical stimulation had little effect on cell survival assessed by trypan blue uptake, CPK release, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining. Electrical stimulation for 24 h did not induce stress response (heat shock protein 70, 90) nor stress kinase (Erk, JNK, p38) activation. NRG stimulation of Erk and Akt was similar between paced and quiescent cells. Pacing sensitized myocytes to beta-AR-stimulated JNK phosphorylation and cell death with 0.1 microM norepinephrine (NE) in paced myocytes causing equivalent cytotoxicity to 10 microM NE in quiescent cells. NRG suppressed beta-AR-induced apoptosis through a phosphatidylinositol-3-kinase-dependent pathway in both paced and quiescent cells, although it is overwhelmed by high-NE concentration in paced cells. Thus myocyte contractility modulates both NE cytotoxicity as well as the cytoprotective effect of NRG. These results demonstrate the feasibility and importance of using electrically paced cardiomyocytes in primary culture when examining the signaling pathways of cell survival. PMID- 14522822 TI - Polymodal regulation of hTREK1 by pH, arachidonic acid, and hypoxia: physiological impact in acidosis and alkalosis. AB - Expression of the human tandem P domain K+ channel, hTREK1, is limited almost exclusively to the central nervous system, where ambient Po2 can be as low as 20 Torr. We have previously shown that this level of hypoxia evokes a maximal inhibitory influence on recombinant hTREK1 and occludes the activation by arachidonic acid; this has cast doubt on the idea that TREK1 activation during brain ischemia could facilitate neuroprotection via hyperpolarizing neurons in which it is expressed. Using both whole cell and cell-attached patch-clamp configurations, we now show that the action of another potent TREK activator and ischemia-related event, intracellular acidification, is similarly without effect during compromised O2 availability. This occlusion is observed in either recording condition, and even the concerted actions of both arachidonic acid and intracellular acidosis are unable to activate hTREK1 during hypoxia. Conversely, intracellular alkalinization is a potent channel inhibitor, and hypoxia does not reverse this inhibition. However, increases in intracellular pH are unable to occlude either arachidonic acid activation or hypoxic inhibition. These data highlight two important points. First, during hypoxia, modulation of hTREK1 cannot be accomplished by parameters known to be perturbed in brain ischemia (increased extracellular fatty acids and intracellular acidification). Second, the mechanism of regulation by intracellular alkalinization is distinct from the overlapping structural requirements known to exist for regulation by arachidonic acid, membrane distortion, and acidosis. Thus it seems likely that hTREK1 regulation in the brain will be physiologically more relevant during alkalosis than during ischemia or acidosis. PMID- 14522823 TI - Niflumic acid inhibits ATP-stimulated exocytosis in a mucin-secreting epithelial cell line. AB - ATP is an efficacious secretagogue for mucin and chloride in the epithelial cell line HT29-Cl.16E. Mucin release has been measured as [3H]glucosamine-labeled product in extracellular medium and as single-cell membrane capacitance increases indicative of exocytosis-related increases in membrane area. The calcium activated chloride channel blocker niflumic acid, also reported to modulate secretion, was used to probe for divergence in the purinergic signaling of mucin exocytosis and channel activation. With the use of whole cell patch clamping, ATP stimulated a transient capacitance increase of 15 +/- 4%. Inclusion of niflumic acid significantly reduced the ATP-stimulated capacitance change to 3 +/- 1%, although normalized peak currents were not significantly different. Ratiometric imaging was used to assess intracellular calcium (Cai2+) dynamics during stimulation. In the presence of niflumic acid, the ATP-stimulated peak change in Cai2+ was unaffected, but the initial response and overall time to Cai2+ peak were significantly affected. Excluding external calcium before ATP stimulation or including the capacitative calcium entry blocker LaCl3 during stimulation muted the initial calcium transient similar to that observed with niflumic acid and significantly reduced peak capacitance change, suggesting that a substantial portion of the ATP-stimulated mucin exocytosis in HT29-Cl.16E depends on a rapid, brief calcium influx through the plasma membrane. Niflumic acid interferes with this influx independent of a chloride channel blockade effect. PMID- 14522824 TI - Activity-dependent regulation of neurohormone synthesis and its impact on reproductive behavior in aplysia. AB - The bag cell neurons (BCNs) of the mollusk Aplysia californica provide a simple model system for investigating cellular and molecular events regulating synthesis and secretion of a reproductive neuropeptide and their impact on physiology and behavior. The BCNs secrete a large amount of egg-laying hormone (ELH) in response to an electrical afterdischarge. The afterdischarge also triggers cellular and molecular events leading to upregulation of ELH biosynthesis to replenish the supply of releasable hormone that was lost because of secretion. In the present review, we discuss signal-transduction events that link membrane excitability to ELH biosynthesis. We present evidence that the afterdischarge stimulates ELH synthesis by upregulating translation of ELH mRNA rather than by activating ELH gene transcription. This increase in ELH synthesis is accompanied by a decrease in total protein synthesis, suggesting that the synthetic machinery is being funneled selectively toward ELH. We also discuss work showing that afterdischarge induced ELH synthesis uses a novel mechanism of translation initiation, one involving a switch from cap-dependent to cap-independent translation initiation that activates an internal ribosome entry site (IRES) located in the 5' untranslated region of ELH mRNA. The IRES-regulated translation provides a unique cellular mechanism to selectively upregulate synthesis of a critical reproductive hormone at the expense of nonessential proteins. PMID- 14522825 TI - Attenuation of estrogenic effects by dihydrotestosterone in the pig uterus is associated with downregulation of the estrogen receptors. AB - Androgens are known to attenuate some effects of estradiol-17beta (E) in the uterus. The objectives of the present experiment were to determine effects of 5alpha-dihydrotestosterone (DHT) on estrogenic actions in the pig uterus and its associations with changes in expression of the estrogen receptor (ER) alpha and ERbeta. Postpubertal gilts (120-130 kg of body weight; n = 16) were ovariectomized, and 3-4 weeks later received once-a-day injections (i.m.) of one of the following treatments during four consecutive days: 1) vehicle (corn oil), 2) E (250 microg), 3) E (250 microg) plus 1 mg DHT, or 4) E (250 microg) plus 10 mg DHT. Uterine tissues were collected 24 h after the last treatment. Gilts receiving E or E plus 1 mg DHT had greater uterine wet weight, uterine horn diameter, luminal epithelium thickness, and endometrial gland diameter compared with gilts treated with vehicle or E plus 10 mg DHT. Gilts receiving E or E plus 1 mg DHT were not different in these characteristics. Relative amounts of mRNAs in the endometrium for the cell proliferation marker histone H2a and the E inducible protein complement component C3 increased in gilts treated with E compared with gilts treated with vehicle. E-induced increases in histone H2a and C3 mRNAs were not altered by cotreatment with E plus 1 mg DHT but were inhibited by E plus 10 mg DHT. Androgen receptor (AR) mRNA in the endometrium increased by treatment with E. Cotreatment of gilts with E and DHT did not alter the E-induced AR mRNA increase. Gilts treated with E plus 10 mg DHT had lesser amounts of immunoreactive ERalpha in cell nuclei of the myometrium and endometrial stroma and a tendency for a decrease in luminal epithelium compared with gilts treated with E. Amounts of immunoreactive ERalpha in glandular epithelium were not influenced by the treatments. Relative amounts of ERalpha and ERbeta mRNAs decreased in the endometrium of gilts treated with E plus 10 mg DHT compared with gilts treated with E. Downregulation of the ERs, particularly ERalpha in the myometrium and endometrial stroma, might be a relevant mechanism in the antagonism of estrogenic effects by DHT in the pig uterus. PMID- 14522826 TI - Protocadherin alpha3 acts at sites distinct from classic cadherins in rat testis and sperm. AB - The testis expresses a variety of cadherin superfamily members including classic cadherins and protocadherins. This report describes the first localization of a protocadherin protein in testis and sperm. After cloning rat cDNAs for protocadherin alpha3 and alpha4, isoform-specific polyclonal antibodies were generated against protocadherin alpha3. Western blotting of rat testis showed that protocadherin alpha3 was solubilized completely by Triton X-100, in contrast to the adhesion junction components N-cadherin, beta-catenin, and p120 catenin. Corroborating this data, protocadherin alpha3 was immunolocalized to the spermatid acrosomal area, intercellular bridge, and flagellum, but not classic cadherin-based adhesion junctions. Acrosome-associated protocadherin alpha3 was first detected at step 8 of spermiogenesis, and this association remained on cauda epididymal sperm. Acrosome immunostaining was reduced, but present, in acrosome-reacted sperm. Spermatid intercellular bridges became positive for protocadherin alpha3 coincident with the appearance of plectin, occurring at spermiogenic steps 8 to 9, and elongate spermatid bridges remained positive throughout spermatogenesis. The developing flagellum was uniformly immunostained for protocadherin alpha3 up to approximately spermiogenic step 17. Subsequently, flagellar immunostaining was confined to the principal piece, and this pattern continued in cauda epididymal sperm. These data show that protocadherin alpha3 performs functions unique from classic cadherins in spermatogenesis and suggest a role for protocadherin alpha3 in organizing germ cell-specific structures including the intercellular bridge, flagellum, and acrosome. PMID- 14522828 TI - Leptin prevents fasting-mediated reductions in pulsatile secretion of luteinizing hormone and enhances its gonadotropin-releasing hormone-mediated release in heifers. AB - We tested the hypothesis that leptin could prevent fasting-mediated reductions in pulsatile secretion and modify GnRH-mediated release of LH in heifers approaching puberty. Thirteen crossbred, prepubertal heifers (13.5-16 mo; 280-350 kg) exhibiting frequencies of pulses of LH between 0.67 and 1 pulse/h, were assigned randomly to two groups: 1). control (n = 6), fasted for 72 h with s.c. injections of saline at 12-h intervals, and 2). leptin (n = 7), fasted for 72 h with s.c. injections of oleptin (19.2 microg/kg) at 12-h intervals. Blood samples were collected intensively for 6 h on Days 0 and 3. This was followed on Day 3 with sequential administration of physiological (0.0011 microg/kg, i.v.) and pharmacological (0.22 microg/kg, i.v.) doses of GnRH and additional blood sampling. Leptin treatment increased (P = 0.0003) plasma concentrations of leptin 5-6-fold compared to controls. Fasting caused a marked decline (P = 0.01) between Days 0 and 3 in the frequency of LH pulses in controls; however, this effect was prevented in the leptin group, with pulse frequency increasing (P < 0.008) from Day 0 to 3. Leptin treatment increased GnRH-induced release of LH at both low (P = 0.04) and high (P = 0.02) doses. Plasma insulin and insulin-like growth factor 1 were reduced by fasting and unaffected by leptin. Leptin increased mean concentrations of growth hormone. Results indicate, for the first time, that exogenous leptin can prevent fasting-mediated reductions in the frequency of LH pulses and modify GnRH-mediated release of LH in intact, prepubertal heifers. PMID- 14522827 TI - Pyrazolo pyrimidine-type inhibitors of SRC family tyrosine kinases promote ovarian steroid-induced differentiation of human endometrial stromal cells in vitro. AB - Reversible protein tyrosine phosphorylation, coordinately controlled by protein tyrosine kinases and phosphatases, is a critical element in signal transduction pathways regulating a wide variety of biological processes, including cell growth, differentiation, and tumorigenesis. We have previously reported that c Src belonging to the Src family tyrosine kinase (SFK) becomes dephosphorylated at tyrosine 530 (Y530) and thereby activated during progestin-induced differentiation of human endometrial stromal cells (i.e., decidualization). In this study, to elucidate the role of decidual c-Src activation, we examined whether 4-amino-5-(4-methylphenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP1) and 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP2), both potent and selective SFK inhibitors, affected the ovarian steroid-induced decidualization in vitro. Unexpectedly, PP1 paradoxically increased the kinase activity of decidual c-Src together with dephosphorylation of Y530 in the presence of ovarian steroids. Concomitantly, PP1 enhanced morphological and functional decidualization, as determined by induction of decidualization markers, such as insulin-like growth factor binding protein-1 and prolactin. PP2 also advanced decidualization along with up-regulation of the active form of c Src whose Y-530 was dephosphorylated. In contrast to PP1 and PP2, herbimycin A, a tyrosine kinase inhibitor with less specificity for SFKs, showed little enhancing effect on the expression of both IGFBP-1 and active c-Src. These results suggest that SFKs, including c-Src, may play a significant role in stromal cell differentiation, providing a clue for a possible therapeutic strategy to modulate endometrial function by targeting signaling pathway(s) involving SFKs. PMID- 14522829 TI - Nitric oxide/nitric oxide synthase, spermatogenesis, and tight junction dynamics. AB - During spermatogenesis, preleptotene and leptotene spermatocytes, residing in the basal compartment of the seminiferous epithelium, must traverse the blood-testis barrier (BTB) to gain entry to the adluminal compartment for further development at late stage VIII and early stage IX of the epithelial cycle. As such, the timely opening and closing of the BTB is crucial to spermatogenesis. A compromise in this process can lead to infertility. Moreover, the BTB is unique in its relative localization in the seminiferous epithelium compared to the tight junctions (TJs) found in other epithelia. Sertoli cell TJs are situated near the basal lamina in the testis, closest to the basement membrane (a modified form of extracellular matrix [ECM]), unlike TJs found in other epithelia, which are found nearest the apical portion of an epithelium, farthest away from ECM. Needless to say, BTB function in the testis is maintained by intricate regulatory mechanisms. In addition to hormones and cytokines, nitric oxide (NO) was recently shown to be a putative TJ regulator in the testis. Perhaps equally important, TJ dynamics in the testis were shown to be regulated, at least in part, by occludin, a TJ integral membrane protein, via the NO/soluble guanylate cyclase/cGMP/protein kinase G signaling pathway. This minireview summarizes recent advances in the field regarding the role of NO in testicular function, with special emphasis regarding its role in TJ dynamics and the likely implications of these studies for male contraceptive development. PMID- 14522830 TI - Phosphorylated endothelial nitric oxide synthase mediates vascular endothelial growth factor-induced penile erection. AB - The objective of the present study was to evaluate whether vascular endothelial growth factor (VEGF)-induced penile erection is mediated by activation of endothelial nitric oxide synthase (eNOS) through its phosphorylation. We assessed the role of constitutively activated eNOS in VEGF-induced penile erection using wild-type (WT) and eNOS-knockout (eNOS(-/-)) mice with and without vasculogenic erectile dysfunction. Adult WT and eNOS(-/-) mice were subjected to sham operation or bilateral castration to induce vasculogenic erectile dysfunction. At the time of surgery, animals were injected intracavernosally with a replication deficient adenovirus expressing human VEGF145 (10(9) particle units) or with empty virus (Ad.Null). After 7 days, erectile function was assessed in response to cavernous nerve electrical stimulation. Total and phosphorylated protein kinase B (Akt) as well as total and phosphorylated eNOS were quantitatively assessed in mice penes using Western immunoblot and immunohistochemistry. In intact WT mice, VEGF145 significantly increased erectile responses, and in WT mice after castration, it completely recovered penile erection. However, VEGF145 failed to increase erectile responses in intact eNOS(-/-) mice and only partially recovered erectile function in castrated eNOS(-/-) mice. In addition, VEGF145 significantly increased phosphorylation of eNOS at Serine 1177 by approximately 2 fold in penes of both intact and castrated WT mice. The data provide a molecular explanation for VEGF stimulatory effect on penile erection, which involves phosphorylated eNOS (Serine 1177) mediation. PMID- 14522831 TI - Expression and activation of protein kinase C isozymes by prostaglandin F(2alpha) in the early- and mid-luteal phase bovine corpus luteum. AB - Western blotting was used to identify the array of protein kinase C (PKC) isozymes expressed in the early (Day 4) and midcycle (Day 10) bovine corpus luteum (CL). PCKalpha, betaI, betaII, epsilon, and micro isozymes were detected in total protein samples prepared from both Day-4 and Day-10 corpora lutea. In contrast, specific antibodies for PKCgamma, eta, lambda, and theta isozymes failed to detect protein bands in the luteal samples. PKCbetaII and epsilon isozymes were expressed differentially at these two developmental stages of the bovine CL. In the Day-4 luteal samples, PKCepsilon was barely detectable; in contrast, in the Day-10 samples, the actin-corrected ratio for PKCepsilon was 1.16 +/- 0.13. This ratio was higher than the detected ratio for PKCbetaI and micro at this developmental phase of the CL (P < 0.01), but it was comparable with the ratio detected for the PCKalpha and betaII. The amount of PKCbetaII was, although not as dramatic, also greater in the Day-10 CL (actin-corrected ratio was 0.85 +/- 0.2) than in the Day-4 CL (0.35 +/- 0.09 [P < 0.01]). The actin corrected ratios for all other PKC isozymes, alpha (Day 4 = 0.93 +/- 0.16, Day 10 = 0.97 +/- 0.09), betaI (Day 4 = 0.54 +/- 0.073, Day 10 = 0.48 +/- 0.74), and micro (Day 4 = 0.21 +/- 0.042, Day 10 = 0.21 +/- 0.38) were not different at these 2 days of the cycle. An experiment was designed to test whether activation of specific isozymes differed between CL that do or do not regress in response to PGF(2alpha). Bovine CL from Day 4 and Day 10 of the estrous cycle were collected and 1 mm CL fragments were treated in vitro for 0, 2.5, 5, 10 or 20 min with PGF(2alpha) (0.1, 1.0, and 10 nM) or minimal essential medium-Hepes vehicle. Translocation of PKC from cytoplasm to membrane fraction was used as indication of PKC activation by PGF(2alpha). Evidence for PKC activation was observed in both Day-4 and Day-10 luteal samples treated with 10 nM PGF(2alpha). Therefore, if PKC, an intracellular mediator associated with the luteal PGF(2alpha) receptor, contributes to the lesser sensitivity of the Day-4 CL, it is likely due to the differential expression of the epsilon and betaII isozymes of PKC at this stage and not due to an inability of the PGF(2alpha) receptor to activate the isozymes expressed in the early CL. PMID- 14522832 TI - Evidence for the regulation of glycosylation of golden hamster (Mesocricetus auratus) oviductin during the estrous cycle. AB - The oviduct contributes to the reproductive environment by secreting various factors, including a family of glycoproteins termed oviductins. Although many studies have demonstrated that ovarian hormones modulate oviductin gene expression in several mammalian species, there has been controversy surrounding the regulation of golden hamster oviductin. The current study was undertaken to investigate the transcriptional and translational modifications of hamster oviductin during the estrous cycle. First, we verified that hamster oviductin mRNA expression remains constant throughout the estrous cycle by semiquantitative reverse transcription polymerase chain reaction. We then developed a polyclonal antibody against recombinant hamster oviductin (rhaOvm). The anti-rhaOvm antibody was subsequently used in conjunction with quantitative immunocytochemistry to investigate the oviductin levels in the hamster oviduct during the estrous cycle. Quantification of immunolabeling revealed a high, consistent level of glycoprotein throughout the estrous cycle. Therefore, it appears that the production of oviductin is not regulated differentially during the estrous cycle. Size variations in hamster oviductin expression were also investigated by Western blot analysis. The oviduct contains several forms of oviductin at each stage of the estrous cycle, the native glycosylated form(s) of 160-350 kDa, and several precursor forms of 70-100 kDa. Although variations in the intensities of the polydispersed band were not evident during the estrous cycle, additional bands ranging from 90 to 100 kDa were detected in the estrus, metestrus, and diestrus 1 stages. The results from the present investigations suggest that whereas ovarian hormones do not appear to influence the hamster oviductin mRNA and protein expressions, glycosylation of hamster oviductin appears to be differentially regulated during the estrous cycle. PMID- 14522833 TI - Nuclear transfer of adult bone marrow mesenchymal stem cells: developmental totipotency of tissue-specific stem cells from an adult mammal. AB - Recent studies have demonstrated that somatic stem cells have a flexible potential greater than previously expected when they are transplanted into different tissues. On the other hand, recent studies also have revealed that these potentials might occur because of spontaneous cell fusion with recipient cells. The nuclei of somatic cells could have been reprogrammed when they were artificially or spontaneously fused with mouse embryonic stem (ES) cells. The resultant hybrid cells acquired a developmental pluripotency that the original somatic cells did not have but that ES cells did. LaBarge and Blau (Cell 2002; 111:589-601) demonstrated that adult bone marrow-derived cells contributed to muscle tissue in a stepwise biological progression. This means that bone marrow derived cells became satellite cells of mononucleate muscle stem cells after the first irradiation-induced damage to the mouse, and after the second irradiation induced damage, multinucleate myofibers appeared from the bone marrow-derived cells. Considered together, the differentiation potential of the somatic stem cell nucleus itself remains unclear. Although the pluripotency of somatic stem cell populations has been evaluated, the developmental totipotency of the nuclei of somatic stem cells, whether or not they fused with other cells, has not been shown, except in only one study concerning fetal neural cells (never in adult stem cells). Here, we showed the developmental totipotency of adult bovine mesenchymal stem cells by nuclear transfer. PMID- 14522834 TI - Expression of platelet-derived growth factor (PDGF) in the epididymis and analysis of the epididymal development in PDGF-A, PDGF-B, and PDGF receptor beta deficient mice. AB - The platelet-derived growth factor (PDGF) family of ligands and receptors play a pivotal role in the development of various organs. The critical importance of the PDGF-mediated signaling during embryonic development and adult physiology of the kidney and the common mesonephric origin of the epididymis and kidney prompted us to investigate the immunohistochemical localization of PDGF A- and B-chain and PDGF receptor (PDGFR) alpha- and beta-subunit in rat and mouse epididymis, the expression profiles of the corresponding mRNAs, and the consequences of a loss-of function mutation at the PDGF-A, PDGF-B, and PDGFR-beta loci on mouse epididymis phenotypic appearance. Prenatally, PDGF-A and PDGFR-alpha immunohistochemical staining was seen in both species, whereas PDGF-B and PDGFR-beta were absent. The cellular localization of PDGF-A within the epithelium and the alpha-receptor in the mesenchyme in either mouse or rat before birth suggests that the PDGF-A/PDGFR alpha system might be involved in the epididymal epithelial-mesenchymal interaction during the fetal period of life. Postnatally, PDGF A- and B-ligand and PDGFR alpha- and beta-subunit were confined in the epithelium. The identity of PDGF and PDGFR proteins were further confirmed by immunoblotting. In line with the immunohistochemical studies, PDGF-A and PDGFR-alpha mRNAs were seen by reverse transcription-polymerase chain reaction in rat and mouse tissue before birth, whereas PDGF-B and PDGFR-beta were almost not detectable. During the first days of life, PDGF-B and PDGFR-beta genes started to appear, and the overall trend in mRNA expression throughout postnatal development showed that the transcripts levels for PDGF-A, PDGF-B, PDGFR-beta, and PDGFR-alpha were constant with the only exception of a progressive decrease of PDGFR-alpha in adult rats. The PDGF-A null mutation strongly influenced the epididymal phenotype starting from puberty; only fetal PDGF-B and PDGFR-beta -/- mice were available, and no differences were seen in the epididymis of these animals, compared with wild-type littermates. Taken together, these data indicate that the PDGF system is highly expressed in the epididymis and suggest that PDGF could be involved in the maintenance of morphological structure and functional control of this organ. PMID- 14522835 TI - Differential regulation of apoptosis in the corpus luteum of pregnancy and newly formed corpus luteum after parturition in rats. AB - Apoptosis contributes to luteal regression in many species. In the postpartum rat, there are two different types of corpora lutea (CL) in the ovary: CL of pregnancy (CLP) and newly formed CL (NCL). To investigate the regulation of apoptosis in the two different types of CL during luteal regression, apoptosis and caspase-3 activity were examined in the CL obtained on Days 7, 15, and 21 of pregnancy and Days 0, 1, 3, 5, 7, and 9 postpartum. Furthermore, the effect of lactation on apoptosis in the CL was examined in two groups of postpartum rats: lactating rats that nurse more than 10 pups, and nonlactating rats that nurse no pups. Apoptotic cells were detected after Day 21 of pregnancy. In the CLP, remarkable increases in the number of apoptotic cells on Days 5 and 9 postpartum were observed in nonlactating rats (P < 0.01), but not in lactating rats. Changes in caspase-3 activity in the CLP were not consistent with those in number of apoptotic cells. In the NCL, an increase in apoptosis was found only on Day 5 postpartum in nonlactating rats (P < 0.01), but not in lactating rats. Changes in caspase-3 activity in the NCL were consistent with those in number of apoptotic cells. In conclusion, apoptosis is, at least in part, involved in luteal regression after parturition, and lactation appears to inhibit apoptosis. This study also suggests the presence of a caspase-3-independent mechanism for apoptosis in CLP regression in the rat. PMID- 14522836 TI - Increased apoptosis occurring during the first wave of spermatogenesis is stage specific and primarily affects midpachytene spermatocytes in the rat testis. AB - The physiological apoptosis that occurs in immature testis appears to be necessary for the maturation of this tissue. Thus, inhibition of the early apoptotic wave associated with the first round of spermatogenesis is followed by accumulation of spermatogonia and infertility later in life. To identify the cell types undergoing apoptosis in immature rat testis and to characterize the relationship between this apoptosis and progression of the first wave of spermatogenesis, sequential viable segments of seminiferous tubules from 8-, 18-, and 26-day-old rats were examined under a phase-contrast microscope. One novel observation was the existence of pronounced stage-specificity during the peak of apoptosis at the very early postnatal ages of 18 and 26 days. Increased apoptosis of pachytene spermatocytes in stages VII-VIII was the major feature that distinguished immature spermatogenesis from the corresponding adult process. The frequency of apoptosis among type A spermatogonia in immature stages IX-I was also elevated in comparison to the corresponding mature stages. The age-related peak of apoptosis was mediated by caspase 3; furthermore, stage-dependent expression of Bax in midpachytene spermatocytes was observed in the 18- and 26 day-old testis. These observations suggest that this Bax-regulated, caspase 3 mediated, increased apoptosis of midpachytene spermatocytes during the first wave of immature spermatogenesis represents a major difference in comparison to apoptosis occurring in the mature testis, and it may play an important regulatory role in establishing spermatogenesis in the rat testis. PMID- 14522837 TI - Depression by relaxin of neurally induced contractile responses in the mouse gastric fundus. AB - The peptide hormone relaxin, which attains high circulating levels during pregnancy, has been shown to depress small-bowel motility through a nitric oxide (NO)-mediated mechanism. In the present study we investigated whether relaxin also influences gastric contractile responses in mice. Female mice in proestrus or estrus were treated for 18 h with relaxin (1 microg s.c.) or vehicle (controls). Mechanical responses of gastric fundal strips were recorded via force displacement transducers. Evaluation of the expression of nitric oxide synthase (NOS) isoforms was performed by immunohistochemistry and Western blot. In control mice, neurally induced contractile responses elicited by electrical field stimulation (EFS) were reduced in amplitude by addition of relaxin to the organ bath medium. In the presence of the NO synthesis inhibitor l-NNA, relaxin was ineffective. Direct smooth muscle contractile responses were not influenced by relaxin or l-NNA. In strips from relaxin-pretreated mice, the amplitude of neurally induced contractile responses was also reduced in respect to the controls, while that of direct smooth muscle contractions was not. Further addition of relaxin to the bath medium did not influence EFS-induced responses, whereas l-NNA did. An increased expression of NOS I and NOS III was observed in gastric tissues from relaxin-pretreated mice. In conclusion, the peptide hormone relaxin depresses cholinergic contractile responses in the mouse gastric fundus by up-regulating NO biosynthesis at the neural level. PMID- 14522838 TI - Role of capsaicin-sensitive nerve fibers in uterine contractility in the rat. AB - The possible participation of capsaicin-sensitive sensory nerves in the modulation of neurogenic contractions was studied in nonpregnant and term pregnant rat uteri. Neurogenic contractions were elicited by electric field stimulation (40 V, 1-70 Hz, 0.6 msec) in intact uteri and uteri that were previously exposed to capsaicin in vitro. In capsaicin pretreated preparations obtained both from nonpregnant and term pregnant rats, a dose-dependent increase in the amplitude of uterine contractions was detected. Prior systemic treatment of the rats with capsaicin (130 mg/kg, s.c.) abolished the effect of in vitro capsaicin administration on the amplitude of neurogenic contractions. Use of a specific antagonist of calcitonin gene-related peptide revealed that depletion of this peptide, which normally elicits uterine smooth muscle relaxation, may be responsible for the increased responsiveness of the uterus to low-frequency stimulation. Experiments on the localization of calcitonin gene-related peptide in uterine tissue specimens exposed to capsaicin revealed dose-dependent depletion of calcitonin-gene related peptide-immunoreactive nerves innervating blood vessels and the myometrium. The findings indicate that capsaicin-sensitive afferent nerves, by the release of sensory neuropeptides, significantly contribute to the modulation of uterine contractility both in nonpregnant and term pregnant rats. It is suggested that uterine sensory nerve activation may be part of a trigger mechanism leading to preterm contractions evoked by, for example, inflammation. PMID- 14522839 TI - Neurons on cannabinoids: dead or alive? PMID- 14522840 TI - Novel data point to a broader mechanism of action of oxidized ATP: the P2X7 receptor is not the only target. AB - Oxidized ATP (oATP) is a Schiff-base-forming reagent that has been used for some years as an antagonist at the P2X7 receptor (P2X7R). Preincubation of mononuclear phagocytes with this inhibitor leads to attenuation of several proinflammatory responses triggered by extracellular ATP as well as a few non-nucleotide agonists. Novel data show that oATP reduces NFkappaB activation and IL-8 release in cells lacking P2X7R, thus suggesting that some anti-inflammatory effects of oATP may not be due to blockade of the P2X7R. This effect of oATP resembles the action of other natural or synthetic Schiff-base-forming reagents with immunomodulatory activity. PMID- 14522842 TI - Oxidized ATP (oATP) attenuates proinflammatory signaling via P2 receptor independent mechanisms. AB - Periodate-oxidized ATP (oATP), which covalently modifies nucleotide-binding proteins, can significantly attenuate proinflammatory signaling. Although the P2X7 nucleotide receptor (P2X7R) is irreversibly antagonized by oATP, it is unclear whether anti-inflammatory actions of oATP are predominantly mediated via its actions on P2X7R. Here, we describe inhibitory effects of oATP on proinflammatory responses in three human cell types that lack expression of P2X7R: human umbilical vein endothelial cells (HUVEC), HEK293 cells, and 1321N1 astrocytes. oATP decreased by 40-70% the secretion of interleukin (IL)-8 stimulated by tumor necrosis factor-alpha (TNF-alpha) in all three cell types, by IL-1beta in HUVEC and 1321N1 cells, and by endotoxin in HUVEC. Attenuation of TNF alpha-stimulated IL-8 secretion by oATP was similar in wild-type HEK cells or HEK cells stably expressing recombinant P2X7R. oATP also attenuated cytokine stimulated expression of nuclear factor-kappaB-luciferase reporter genes expressed in HEK or 1321N1 cells, but did not affect the rapid downregulation of IkappaB. oATP had no effect on uridine triphosphate-induced activation of native P2Y2 receptors in HEK cells, but reduced the potency and efficacy of ADP as an agonist of native P2Y1 receptors. However, inhibition of P2Y1 receptors with the specific antagonist MRS2216 did not mimic the effects of oATP on TNF-alpha stimulated IL-8 secretion. Although 1321N1 astrocytes lack expression of any known P2 receptor subtypes, oATP markedly inhibited ecto-ATPase activity in these cells, resulting in a significant accumulation of extracellular ATP. In summary, oATP can attenuate proinflammatory signaling by mechanisms independent of the expression or activation of known P2 receptor subtypes. PMID- 14522843 TI - Tetrahydrocannabinol-induced neurotoxicity depends on CB1 receptor-mediated c-Jun N-terminal kinase activation in cultured cortical neurons. AB - Delta9-Tetrahydrocannabinol (THC), the main psychoactive ingredient of marijuana, induces apoptosis in cultured cortical neurons. THC exerts its apoptotic effects in cortical neurons by binding to the CB1 cannabinoid receptor. The CB1 receptor has been shown to couple to the stress-activated protein kinase, c-Jun N-terminal kinase (JNK). However, the involvement of specific JNK isoforms in the neurotoxic properties of THC remains to be established. The present study involved treatment of rat cultured cortical neurons with THC (0.005-50 microM), and combinations of THC with the CB1 receptor antagonist, AM 251 (10 microM) and pertussis toxin (PTX; 200 ng ml-1). Antisense oligonucleotides (AS) were used to deplete neurons of JNK1 and JNK2 in order to elucidate their respective roles in THC signalling. Here we report that THC induces the activation of JNK via the CB1 receptor and its associated G-protein, Gi/o. Treatment of cultured cortical neurons with THC resulted in a differential timeframe of activation of the JNK1 and JNK2 isoforms. Use of specific JNK1 and JNK2 AS identified activation of caspase-3 and DNA fragmentation as downstream consequences of JNK1 and JNK2 activation. The results from this study demonstrate that activation of the CB1 receptor induces JNK and caspase-3 activation, an increase in Bax expression and DNA fragmentation. The data demonstrate that the activation of both JNK1 and JNK2 isoforms is central to the THC-induced activation of the apoptotic pathway in cortical neurons. PMID- 14522841 TI - On the selectivity of superoxide dismutase mimetics and its importance in pharmacological studies. AB - The list of pathophysiological conditions associated with the overproduction of superoxide expands every day. Much of the knowledge compiled on the role of this radical in disease has been gathered using the native superoxide dismutase enzyme and, more recently, by the use of superoxide dismutase knockout models or transgenic models that overexpress the various isoforms of the enzyme. Although the native enzyme has shown promising anti-inflammatory properties in both preclinical and clinical studies, there were drawbacks and issues associated with its use as a therapeutic agent and pharmacological tool. Based on the concept that removal of superoxide modulates the course of inflammation, synthetic, low molecular-weight mimetics of the superoxide dismutase enzymes that could overcome some of the limitations associated with the use of the native enzyme have been designed. In this review, we will discuss the advances made using various superoxide dismutase mimetics that led to the proposal that superoxide (and/or the product of its interaction with nitric oxide, peroxynitrite) is an important mediator of inflammation, and to the conclusion that superoxide dismutase mimetics can be utilized as therapeutic agents in diseases of various etiologies. The importance of the selectivity of such compounds in pharmacological studies will be discussed. PMID- 14522844 TI - Electrophysiological evidence for a gradient of G protein-gated K+ current in adult mouse atria. AB - Whole cell current and voltage clamp techniques were used to examine the properties of acetylcholine-sensitive K+ current (IKACh) in myocytes from adult mouse atrium. Superfusion of a maximal dose of carbachol (CCh; 10 microM) caused a substantial increase in K+ current in all myocytes examined. The current voltage (I-V) relation of maximally activated IKACh exhibited weak inward rectification. Consequently, CCh increased the amount of depolarising current necessary to evoke action potentials (APs), and APs evoked in CCh had significantly shorter durations than control APs (P<0.05). The effects of CCh on K+ current and on AP properties were blocked by the muscarinic receptor antagonist methoctramine (1 microM). ACh (10 microM) activated a K+ current with identical properties to that activated by CCh, as did the A1 receptor agonist adenosine (100 microM). Right atrial myocytes had significantly more IKACh than left atrial myocytes (P<0.05), regardless of whether IKACh was evoked by superfusion of muscarinic or A1 receptor agonists. IKACh current density was significantly higher in SA node myocytes than either right or left atrial myocytes. These data identify a gradient of IKACh current density across the supraventricular structures of mouse heart. This gradient, combined with the heterogeneous distribution of parasympathetic innervation of the atria, may contribute to the proarrhythmic ability of vagal nerve stimulation to augment dispersion of atrial refractoriness. PMID- 14522845 TI - Introduction to the transmissible spongiform encephalopathies or prion diseases. AB - Sheep scrapie has been known for at least 200 years and was described as a transmissible disease over 100 years ago. Since then, three groups of transmissible spongiform encephalopathies or TSE diseases have been identified in humans including familial, infectious and sporadic types. The discovery of the prion protein (PrP) in the 1980s greatly accelerated knowledge of the biology and pathogenesis of TSE diseases as this protein was found to play a critical role in disease susceptibility and the TSE species-barrier and may also be a component of the infectious agent itself. Nevertheless, the nature of the TSE agents remains an enigma. Proof of the protein-only hypothesis may require generation of biologically active transmissible agent in a cell-free environment where a virus cannot replicate. Conversely, proof of a viral aetiology will require identification and isolation of a candidate virus. Further understanding of the structure of the disease-associated protease-resistant PrP should help elucidate the mechanism of PrP conversion from the normal to the abnormal form. Such information should open up new approaches to both diagnosis and therapy. PMID- 14522846 TI - Biochemistry and structure of PrP(C) and PrP(Sc). AB - In a brief historical description, it is shown that the prion model was developed from the biochemical and biophysical properties of the scrapie infectious agent. The biochemical properties of the prion protein which is the major, if not only, component of the prion are outlined in detail. PrP is a host-encoded protein which exists as PrP(C) (cellular) in the non-infected host, and as PrP(Sc) (scrapie) as the major component of the scrapie infectious agent. An overview of the purification techniques is given. Although chemically identical, the biophysical features of PrP(Sc) are drastically different in respect to solubility, structure, and stability; furthermore, specific lipids and a polyglucose scaffold were found in prions, whereas for nucleic acids their absence could be proven. The structure of recombinant PrP in solution is known from spectroscopic studies and with high resolution from NMR analysis. Structural models of PrP(Sc) were derived recently from electron microscopic analysis of two dimensional crystals. Conformational transitions of PrP in vitro were studied with different techniques in order to mimic the natural PrP(C) to PrP(Sc) conversion. Spontaneous transitions can be induced by solvent changes, but at present infectivity cannot be induced in vitro. PMID- 14522847 TI - Physiological and pathological functions of the prion protein homologue Dpl. AB - A misfolded version of the prion protein PrP(C), known as PrP(Sc), is the major component of scrapie infectivity, the pathological agent in transmissible spongiform encephalopathies. The Prnp gene that encodes the cellular PrP(C) protein was cloned almost 20 years ago, but remained without sequence or structural relatives for over a decade. Only recently a novel protein, named Doppel (Dpl), was identified, which shares significant biochemical and structural homology with PrP(C). When overexpressed, Dpl is neurotoxic and causes a neurological disease. Strikingly, Dpl neurotoxicity is counteracted and prevented by PrP(C). In contrast to its homologue PrP(C), Dpl is dispensable for prion disease progression and for the generation of PrP(Sc), but Dpl appears to have an essential function in male spermatogenesis. Although Dpl research is still in its infancy, the discovery of Dpl has already solved some enigmas of prion biology and an understanding of its physiological function is emerging. PMID- 14522848 TI - PrP knock-out and PrP transgenic mice in prion research. AB - Spongiform encephalopathies such as scrapie in sheep, bovine spongiform encephalopathy (BSE) in cattle or Creutzfeldt-Jacob disease (CJD) and Gerstmann Straussler-Scheinker syndrome (GSS) in humans is caused by a transmissible agent designated prion. The 'protein only' hypothesis proposes that the prion consists partly or entirely of a conformational isoform of the normal host protein PrP(C), designated PrP(*)(1) and that the abnormal conformer, when introduced into the organism, causes the conversion of PrP(C) into a likeness of itself. PrP(*) may be congruent with PrP(Sc), a protease-resistant, aggregated conformer of PrP that accumulates mainly in brain of almost all prion-infected organisms. PrP(C) consists of a flexible N-terminal half, comprising Cu(2+)-binding octapeptide repeats, and a globular domain consisting of three alpha-helices, one short antiparallel beta-sheet and a single disulphide bond. It is anchored at the outer cell-surface by a glycosyl phosphatidylinositol (GPI) tail and is present in almost all tissues, however, mainly in brain. Compelling linkage between the prion and PrP was established by biochemical and genetic data and led to the prediction that animals devoid of PrP should be resistant to experimental scrapie and fail to propagate infectivity. This prediction was indeed borne out, adding substantial support to the 'protein only' hypothesis. In addition, the availability of PrP knock-out mice provided an approach to carry out reverse genetics on PrP, both in regard to prion disease and to its physiological role. PMID- 14522849 TI - Putative functions of PrP(C). AB - While the exact function of the cellular prion protein (PrP(C)) remains unknown, there are several leads due to increasing knowledge on the localisation and interaction of PrP(C) with other molecules. This chapter will concentrate on these aspects. Identified ligands of PrP(C) mainly belong to the categories of heat-shock proteins, membrane-bound receptors, or heparan sulphates. The possible synaptic role of PrP(C) has been exemplified by electrophysiological findings in PrP(o/o) mice and the studies of PrP(C) as a copper-binding molecule that could regulate the copper content of the synaptic cleft. The latter property of PrP(C) may also endow PrP(C) with the activity of a copper-dependent superoxide dismutase. Binding of PrP(C) to signalling molecules suggests a role as a transmitter of information from the extracellular milieu to the cell and a trigger for a molecular cascade. This agrees with new data on PrP(C) receptors and the role of PrP(C) in cell survival. PMID- 14522850 TI - Trafficking, turnover and membrane topology of PrP. AB - Cell biological studies of PrP have contributed enormously to our understanding of prion diseases. Like other membrane proteins, PrP(C) is post-translationally processed in the endoplasmic reticulum and Golgi on its way to the cell surface after synthesis. Cell surface PrP(C) constitutively cycles between the plasma membrane and early endosomes via a clathrin-dependent mechanism, a pathway consistent with a suggested role for PrP(C) in cellular trafficking of copper ions. PrP molecules carrying mutations linked to inherited prion diseases display several abnormalities in their biochemical properties, maturation, and localisation that may explain their pathogenicity. Recent results have clarified the role of the proteasome in degradation of PrP, and the properties of a transmembrane form of PrP which may play a neurotoxic role in prion diseases. PMID- 14522851 TI - Prion propagation in cultured cells. AB - Cell cultures represent relevant and useful experimental models of transmissible spongiform encephalopathies (TSEs). They are able to promote, upon subpassaging, stable and persistent replication of PrP(Sc) as well as infectivity. To date, only a few cell culture models permissive to prion replication are available. Among them, mouse neuroblastoma cell lines (N2a) are most commonly used. While they correspond to homologous models supporting propagation of mouse-adapted scrapie strains, recent studies have reported heterologous models sensitive to natural occurring disease. Infected cell culture models have provided some valuable insights into the biogenesis of PrP(Sc) in terms of conversion, subcellular localisations, physiopathological consequences and species-barrier determinants. They have also contributed to the screening and the study of possible therapeutic compounds and to the development of new strategies for the investigation of TSE-specific diagnostic markers. PMID- 14522852 TI - TSE strain variation. AB - Studies in mice have revealed considerable strain variation in the agents causing transmissible spongiform encephalopathies (TSEs). TSE strains interact with genetic factors in the host (in particular PrP genotype) to influence characteristics of the disease such as incubation period and neuropathology. TSE strains can retain their identity after propagation in different host species or PrP genotypes, showing that these agents carry their own strain-specific information. It is not known whether this information resides in specific self perpetuating modifications of PrP, or whether a separate informational molecule is required. Strain typing in mice can be used to explore links between TSEs occurring naturally in different species. Such studies have demonstrated that the strain causing BSE in cattle has also infected domestic cats and exotic ungulates. Most importantly, the BSE strain has also been isolated from patients with variant CJD. In contrast, different TSE strains are associated with sporadic CJD and sheep scrapie. PMID- 14522854 TI - Neuropathology of prion diseases. AB - In prion diseases, neuropathology has remained the most important tool to give a definite diagnosis, and neuropathological research has contributed significantly to our current pathogenetic understanding. Immunohistochemistry for the disease associated prion protein (PrP(Sc)) is indispensable for the neuropathological confirmation of prion diseases. The amount and distribution of PrP(Sc) deposits do not always correlate with type and severity of local tissue damage. PrP(Sc) deposition occurs only where neuronal parenchyma is present; in scarred infarctions with prominent gliosis, PrP(Sc) does not accumulate. Early, severe and selective loss affects a subset of inhibitory GABAergic neurons both in human and experimental prion diseases. The central pathogenetic cascade includes oxidative stress to neurons and their apoptosis. New patterns of PrP(Sc) immunoreactivity include granular ganglionic and tiny adaxonal PrP(Sc) deposits in peripheral nervous tissue, and dendritic cells and macrophages in vessel walls, suggesting that mobile haematogenous cells may be involved in spread of prions. PMID- 14522853 TI - Prion protein conversions: insight into mechanisms, TSE transmission barriers and strains. AB - Conversion of PrP(C) to aberrant forms such as PrP(Sc) appears to be critical in the transmission and pathogenesis of transmissible spongiform encephalopathies (TSEs) or prion diseases. In vitro studies have shown that TSE-associated, protease-resistant forms of PrP can cause PrP(C) to convert to forms that are similarly protease-resistant under a wide variety of conditions. These observations have provided evidence that pathological forms of PrP have at least limited capacity to propagate themselves, which is necessary for them to be infectious. PrP conversion reactions have proven to be highly specific and appear to account, at least in part, for TSE species barriers and the propagation of strains. Such in vitro conversion systems have yielded insights into the molecular mechanisms of TSE disease and are being exploited as screens for anti TSE drugs and as bases for diagnostic tests. PMID- 14522855 TI - CNS pathogenesis of prion diseases. AB - Prion diseases or transmissible spongiform encephalopathies (TSEs) are fatal neurodegenerative diseases, clinically characterised by cognitive decline, paralleled by severe damage to the central nervous system. Prion diseases have attracted a broad interest because of their unique mechanisms of replication and propagation; however, the underlying pathogenic mechanisms are still highly speculative. In this review, current knowledge about the pathogenesis of prion diseases in the CNS will be highlighted and the most revealing animal models will be discussed, with future perspectives to address immediate questions about the pathogenesis. PMID- 14522856 TI - Immune system and peripheral nerves in propagation of prions to CNS. AB - Prions are not only unique in the way they replicate. Also the sequence of events triggered by peripheral prion infection, generically termed 'peripheral pathogenesis', sets prions aside from all other known pathogens. Whereas most bacteria, parasites, and viruses trigger innate and adaptive immune responses, the mammalian immune system appears to be remarkably oblivious to prions. Transmissible spongiform encephalopathies (TSEs) do not go along with inflammatory infiltrates, and antibodies to the prion protein are not typically raised during the course of the disease. On the other hand, there is conspicuous involvement of lymphoid organs, which accumulate sizeable concentrations of the infectious agent early during disease. Moreover, various states of immune deficiency can abolish peripheral pathogenesis and prevent 'take' of infection when prions are administered to peripheral sites. Here, we critically re-visit the current evidence for an involvement of the immune system in prion diseases, and will attempt to trace the elaborate mechanisms by which prions, upon entry into the body from peripheral sites, reach the brain. PMID- 14522857 TI - Subclinical prion infection in humans and animals. AB - Transmission of prion diseases between mammalian species is limited by a so called 'species' or 'transmission' barrier. Recognition of prion transmission usually relies on the appearance of clinical symptoms in inoculated animals and the interval between inoculation and appearance of clinical disease is designated incubation period. At some point during this clinically silent period, neuropathological and biochemical changes as well as accumulation of prions in the brain can be detected and this stage can be called preclinical prion disease. Recently, several lines of evidence have suggested that subclinical forms of prion disease exist, in which high levels of infectivity and PrP(Sc) are found in animals that do not develop clinically apparent disease during a normal life span. Such asymptomatic prion 'carrier' states challenge our current understanding of pathogenesis as well as of the molecular basis of barriers to transmission. Subclinical as well as preclinical/clinical prion disease may be relevant when analysing the risk to public health of potential sources of prion exposure. PMID- 14522858 TI - Scrapie and experimental BSE in sheep. AB - Scrapie is a natural disease of sheep, but it can also be successfully transmitted between sheep by experimental inoculation. Although BSE is primarily a disease of cattle, it has also infected humans (causing vCJD) and, in addition, can be transmitted orally to sheep bringing concerns that BSE might naturally have infected the UK sheep population. Because of this, scrapie and BSE are being compared and studied in detail in sheep. PrP genotype controls sheep susceptibility and resistance to scrapie and to BSE, and deposition of the disease-associated PrP(Sc), used as a marker of infection, has the potential to act as a means of identifying BSE-infected animals and describing different pathogenesis mechanisms. Sheep orally dosed with BSE show signs of infection in their blood and this model is of major importance in the study of the safety of blood products for use with human beings. PMID- 14522859 TI - Bovine spongiform encephalopathy (BSE) and its epidemiology. AB - Since the recognition of BSE in 1986, over 180,000 cattle in the UK have developed the disease and 1-3 million are likely to have been infected with the BSE agent, most of which were slaughtered for human consumption before developing signs of the disease. The origin of the first case of BSE is unknown, but the epidemic was caused by the recycling of processed waste parts of cattle, some of which were infected with the BSE agent, to other cattle in feed. Control measures have resulted in the consistent decline of the epidemic in the UK since 1992. Cattle and feed exported from the UK have seeded smaller epidemics in other European countries, where control measures were applied later. If the control measures now in place to protect public and animal health are well enforced, the epidemic in cattle should be largely under control and any remaining risk to humans through the consumption of beef should be very small. PMID- 14522860 TI - Other animal prion diseases. AB - In addition to bovine spongiform encephalopathy (BSE) of cattle and scrapie of sheep and goats, a few other animal prion diseases have been reported. These include feline spongiform encephalopathy of zoological and domestic cats (FSE) and transmissible spongiform encephalopathy (TSE) of zoological ruminants and non human primates, as well as chronic wasting disease of deer and elk (CWD) and transmissible mink encephalopathy of farmed mink (TME). The origins of TSE in cats, zoo bovids, and non-human primates are clearly linked to the BSE epidemic; however, the origins of CWD and TME are less clear, but are not epidemiologically linked to the BSE epidemic. Here we review the epidemiology, transmission, clinical features and pathology of these other animal prion diseases. PMID- 14522861 TI - Sporadic and familial CJD: classification and characterisation. AB - Prion diseases are unique transmissible neurodegenerative diseases that have diverse phenotypes and can be familial, sporadic, or acquired by infection. Recent findings indicate that the PrP genotype and the PrP(Sc) type have a major influence on the disease phenotype in both sporadic and familial human prion diseases. This review attempts to classify and characterise sporadic and familial Creutzfeldt-Jakob disease (CJD) as a function of these two disease determinants. Based on the genotype at codon 129 on both PRNP alleles, the size of protease resistant PrP(Sc) fragments and disease phenotype, we divide sporadic CJD into six subtypes: sCJDMM1/sCJDMV1, sCJDVV2, sCJDMV2, sCJDMM2, sCJDVV1, and sporadic fatal insomnia (sFI). Familial CJD is classified into many haplotypes based on the PRNP mutation and codon 129 (and other polymorphic codons) on the mutant allele. The clinical and pathological features are summarised for each sporadic CJD subtype and familial CJD haplotype. PMID- 14522862 TI - Molecular and clinical classification of human prion disease. AB - While rare in humans, the prion diseases have become an area of intense clinical and scientific interest. The recognition that variant Creutzfeldt-Jakob disease is caused by the same prion strain as bovine spongiform encephalopathy in cattle has dramatically highlighted the need for a precise understanding of the molecular biology of human prion diseases. Detailed clinical, pathological and molecular data from a large number of human prion disease cases have shown that distinct abnormal isoforms of prion protein are associated with prion protein gene polymorphism and neuropathological phenotypes. A molecular classification of human prion diseases seems achievable through characterisation of structural differences of the infectious agent itself. PMID- 14522863 TI - Acquired prion disease: iatrogenic CJD, variant CJD, kuru. AB - Human prion diseases can be classified as sporadic, hereditary or acquired. The cause of sporadic Creutzfeldt-Jakob disease (CJD) is unknown, hereditary cases are associated with mutations of the prion protein gene (PRNP) and acquired forms are caused by the transmission of infection from human to human or, as a zoonosis, from cattle to human. Although acquired forms of human prion disease are rare, the transmission of a fatal and untreatable neurological disorder has had major implications for public health and public policy. PMID- 14522864 TI - Diagnosis of prion diseases. AB - Prion diseases are usually diagnosed clinically and confirmed by post-mortem histopathological examination of brain tissue. The only reliable molecular marker for prion diseases is PrP(Sc), the pathological conformer of the prion protein that accumulates in the central nervous system and, to a lesser extent, in lymphoreticular tissues. For BSE, several commercial diagnostic kits based on the post-mortem immunochemical detection of PrP(Sc) in brain tissue are now available. These rapid screening tests have been used in active surveillance of BSE and have greatly improved the detection of infected cattle before their entry into the human food chain. At present, no diagnostic test exists for the detection of prion diseases in live animals or humans. New diagnostic techniques aimed at increasing sensitivity and specificity of PrP(Sc) detection in body fluids and at identifying novel surrogate markers are under development. In this report, we review the classical diagnostic methods as well as present and future tools for the diagnosis of prion diseases. PMID- 14522865 TI - Approaches to prophylaxis and therapy. AB - Despite important progress in experimental treatment of neurodegenerative diseases, no therapeutic strategy has today proven its capability to cure or even to stabilise human TSEs. Pathogenesis experiments performed in rodent TSE models have shown that central nervous system damages are detectable long before the appearance of the clinical symptoms. At the time of disease onset, PrP(Sc) accumulation has almost reached its highest level, and the neuropathological lesions (spongiosis, gliosis) are as intense as they are at the time of death. Therefore, the neurodegeneration that is present at the onset of the disease is beyond therapy, and, in theory, only a preclinical diagnosis of TSEs would permit the prevention (or delay) of neurodegeneration. Unfortunately, there are no diagnostic tests that can be used to show TSE agent infection during the preclinical phase of the disease. Nevertheless, since the appearance of variant Creutzfeldt-Jakob disease (vCJD), those in the scientific community working on experimental therapy have increased their efforts. Tens of drugs have been tested in several experimental models, and there are some high-output screening platforms being used in Europe and in the US. Any rational therapeutic strategy needs to be based on pathogenesis data and/or knowledge on the nature of the causative agent. Therefore, progress in therapy is tightly linked to a better understanding of the basic science of TSE. PMID- 14522866 TI - Preventing accidental transmission of human transmissible spongifom encephalopathies. AB - The mechanism by which humans became infected with the BSE agent is discussed, and the matter of potential person-to person transmission of TSEs through contagion or medical procedures is considered. There is some discussion regarding the current evidence relating to whether or not the blood of humans infected with TSEs is infectious. Considerable emphasis is placed on the fact that TSE agents are known to be relatively resistant to decontamination by procedures that are effective with conventional micro-organisms, including (under worst-case conditions) the autoclaving procedures used to sterilise surgical instruments. Methods for providing additional re-assurance with regard to the safety of instruments are described. Safety in the pathology laboratory is discussed extensively because TSE agents are not inactivated by the usual processes used to fix tissues, and such laboratories will receive fixed tissues that are still highly infectious as far as TSE agents are concerned. PMID- 14522867 TI - Ethical issues in human prion diseases. AB - Prion diseases or transmissible spongiform encephalopathies are a group of closely related transmissible neurodegenerative conditions of humans and animals, all of which are incurable. In recent years, they have captured public attention with the emergence of the bovine spongiform encephalopathy (BSE) epidemic in Europe, and more recently with the appearance of variant CJD (vCJD) in humans, a novel form of Creutzfeldt-Jakob disease (CJD) that is linked to dietary exposure to BSE. In this chapter, we outline ethical questions posed by research, diagnostic procedures and therapy in the field of prion diseases. PMID- 14522868 TI - Wnt signaling is required at distinct stages of development for the induction of the posterior forebrain. AB - One of the earliest manifestations of anteroposterior pattering in the developing brain is the restricted expression of Six3 and Irx3 in the anterior and posterior forebrain, respectively. Consistent with the role of Wnts as posteriorizing agents in neural tissue, we found that Wnt signaling was sufficient to induce Irx3 and repress Six3 expression in forebrain explants. The position of the zona limitans intrathalamica (zli), a boundary-cell population that develops between the ventral (vT) and dorsal thalamus (dT), is predicted by the apposition of Six3 and Irx3 expression domains. The expression patterns of several inductive molecules are limited by the zli, including Wnt3, which is expressed posterior to the zli in the dT. Wnt3 and Wnt3a were sufficient to induce the dT marker Gbx2 exclusively in explants isolated posterior to the presumptive zli. Blocking the Wnt response allowed the induction of the vT-specific marker Dlx2 in prospective dT tissue. Misexpression of Six3 in the dT induced Dlx2 expression and inhibited the expression of both Gbx2 and Wnt3. These results demonstrate a dual role for Wnt signaling in forebrain development. First, Wnts directed the initial expression of Irx3 and repression of Six3 in the forebrain, delineating posterior and anterior forebrain domains. Later, continued Wnt signaling resulted in the induction of dT specific markers, but only in tissues that expressed Irx3. PMID- 14522869 TI - PP2A:B56epsilon is required for Wnt/beta-catenin signaling during embryonic development. AB - The Wnt/beta-catenin pathway plays important roles during embryonic development and growth control. The B56 regulatory subunit of protein phosphatase 2A (PP2A) has been implicated as a regulator of this pathway. However, this has not been investigated by loss-of-function analyses. Here we report loss-of-function analysis of PP2A:B56epsilon during early Xenopus embryogenesis. We provide direct evidence that PP2A:B56epsilon is required for Wnt/beta-catenin signaling upstream of Dishevelled and downstream of the Wnt ligand. We show that maternal PP2A:B56epsilon function is required for dorsal development, and PP2A:B56epsilon function is required later for the expression of the Wnt target gene engrailed, for subsequent midbrain-hindbrain boundary formation, and for closure of the neural tube. These data demonstrate a positive role for PP2A:B56epsilon in the Wnt pathway. PMID- 14522870 TI - Axolotl pronephric duct migration requires an epidermally derived, laminin 1 containing extracellular matrix and the integrin receptor alpha6beta1. AB - The epidermis overlying the migrating axolotl pronephric duct is known to participate in duct guidance. This epidermis deposits an extracellular matrix onto the migrating duct and its pathway that is a potential source of directional guidance cues. The role of this matrix in pronephric duct guidance was assayed by presenting matrix deposited on microcarriers directly to migrating pronephric ducts in situ. We found that reorientation of extracellular-matrix-bearing carriers prior to their presentation to migrating ducts caused a corresponding reorientation of pronephric duct migration. Subepidermal microinjection of function-blocking antibodies against alpha6 integrin, beta1 integrin or the laminin-1/E8 domain recognized by alpha6beta1 integrin, all of which were detected and localized here, inhibited pronephric duct migration. Moreover, pre exposure to anti-laminin-1/E8 function-blocking antibody prevented reoriented carriers of epidermally deposited matrix from reorienting pronephric duct migration. These results are incorporated into an integrated model of pronephric duct guidance consistent with all present evidence, proposing roles for the previously implicated glial cell-line derived neurotrophic factor and its receptor as well as for laminin 1 and alpha6beta1 integrin. PMID- 14522871 TI - Genetic analysis of adventitious root formation with a novel series of temperature-sensitive mutants of Arabidopsis thaliana. AB - When cultured on media containing the plant growth regulator auxin, hypocotyl explants of Arabidopsis thaliana generate adventitious roots. As a first step to investigate the genetic basis of adventitious organogenesis in plants, we isolated nine temperature-sensitive mutants defective in various stages in the formation of adventitious roots: five root initiation defective (rid1 to rid5) mutants failed to initiate the formation of root primordia; in one root primordium defective (rpd1) mutant, the development of root primordia was arrested; three root growth defective (rgd1, rgd2, and rgd3) mutants were defective in root growth after the establishment of the root apical meristem. The temperature sensitivity of callus formation and lateral root formation revealed further distinctions between the isolated mutants. The rid1 mutant was specifically defective in the reinitiation of cell proliferation from hypocotyl explants, while the rid2 mutant was also defective in the reinitiation of cell proliferation from root explants. These two mutants also exhibited abnormalities in the formation of the root apical meristem when lateral roots were induced at the restrictive temperature. The rgd1 and rgd2 mutants were deficient in root and callus growth, whereas the rgd3 mutation specifically affected root growth. The rid5 mutant required higher auxin concentrations for rooting at the restrictive temperature, implying a deficiency in auxin signaling. The rid5 phenotype was found to result from a mutation in the MOR1/GEM1 gene encoding a microtubule associated protein. These findings about the rid5 mutant suggest a possible function of the microtubule system in auxin response. PMID- 14522872 TI - The beta-catenin/VegT-regulated early zygotic gene Xnr5 is a direct target of SOX3 regulation. AB - In Xenopus laevis, beta-catenin-mediated dorsal axis formation can be suppressed by overexpression of the HMG-box transcription factor XSOX3. Mutational analysis indicates that this effect is due not to the binding of XSOX3 to beta-catenin nor to its competition with beta-catenin-regulated TCF-type transcription factors for specific DNA binding sites, but rather to SOX3 binding to sites within the promoter of the early VegT- and beta-catenin-regulated dorsal-mesoderm-inducing gene Xnr5. Although B1-type SOX proteins, such as XSOX3, are commonly thought to act as transcriptional activators, XSOX3 acts as a transcriptional repressor of Xnr5 in both the intact embryo and animal caps injected with VegT RNA. Expression of a chimeric polypeptide composed of XSOX3 and a VP16 transcriptional activation domain or morpholino-induced decrease in endogenous XSOX3 polypeptide levels lead to an increase in Xnr5 expression, as does injection of an anti-XSOX3 antibody that inhibits XSOX3 DNA binding. These observations indicate that maternal XSOX3 acts in a novel manner to restrict Xnr5 expression to the vegetal hemisphere. PMID- 14522873 TI - Neuronal defects in the hindbrain of Hoxa1, Hoxb1 and Hoxb2 mutants reflect regulatory interactions among these Hox genes. AB - Hox genes are instrumental in assigning segmental identity in the developing hindbrain. Auto-, cross- and para-regulatory interactions help establish and maintain their expression. To understand to what extent such regulatory interactions shape neuronal patterning in the hindbrain, we analysed neurogenesis, neuronal differentiation and motoneuron migration in Hoxa1, Hoxb1 and Hoxb2 mutant mice. This comparison revealed that neurogenesis and differentiation of specific neuronal subpopulations in r4 was impaired in a similar fashion in all three mutants, but with different degrees of severity. In the Hoxb1 mutants, neurons derived from the presumptive r4 territory were re specified towards an r2-like identity. Motoneurons derived from that territory resembled trigeminal motoneurons in both their migration patterns and the expression of molecular markers. Both migrating motoneurons and the resident territory underwent changes consistent with a switch from an r4 to r2 identity. Abnormally migrating motoneurons initially formed ectopic nuclei that were subsequently cleared. Their survival could be prolonged through the introduction of a block in the apoptotic pathway. The Hoxa1 mutant phenotype is consistent with a partial misspecification of the presumptive r4 territory that results from partial Hoxb1 activation. The Hoxb2 mutant phenotype is a hypomorph of the Hoxb1 mutant phenotype, consistent with the overlapping roles of these genes in facial motoneuron specification. Therefore, we have delineated the functional requirements in hindbrain neuronal patterning that follow the establishment of the genetic regulatory hierarchy between Hoxa1, Hoxb1 and Hoxb2. PMID- 14522874 TI - Mixer/Bon and FoxH1/Sur have overlapping and divergent roles in Nodal signaling and mesendoderm induction. AB - Transcription factors belonging to the FoxH1 and Mixer families are required for facets of Nodal signaling during vertebrate mesendoderm induction. Here, we analyze whether zebrafish proteins related to FoxH1 [Schmalspur (Sur)] and Mixer [Bonnie and clyde (Bon)] act within or downstream of the Nodal signaling pathway, test whether these two factors have additive or overlapping activities, and determine whether FoxH1/Sur and Mixer/Bon can account for all Nodal signaling during embryogenesis. We find that sur expression is independent of Nodal signaling and that bon is expressed in the absence of Nodal signaling but requires Nodal signaling and Sur for enhanced, maintained expression. These results and the association of FoxH1 and Mixer/Bon with phosphorylated Smad2 support a role for these factors as components of the Nodal signaling pathway. In contrast to the relatively mild defects observed in single mutants, loss of both bon and sur results in a severe phenotype characterized by absence of prechordal plate, cardiac mesoderm, endoderm and ventral neuroectoderm. Analysis of Nodal regulated proteins reveals that Bon and Sur have both distinct and overlapping regulatory roles. Some genes are regulated by both Bon and Sur, and others by either Bon or Sur. Complete loss of Nodal signaling results in a more severe phenotype than loss of both Bon and Sur, indicating that additional Smad associated transcription factors remain to be identified that act as components of the Nodal signaling pathway. PMID- 14522875 TI - The Caenorhabditis elegans nonmuscle myosin genes nmy-1 and nmy-2 function as redundant components of the let-502/Rho-binding kinase and mel-11/myosin phosphatase pathway during embryonic morphogenesis. AB - Rho-binding kinase and the myosin phosphatase targeting subunit regulate nonmuscle contractile events in higher eukaryotes. Genetic evidence indicates that the C. elegans homologs regulate embryonic morphogenesis by controlling the actin-mediated epidermal cell shape changes that transform the spherical embryo into a long, thin worm. LET-502/Rho-binding kinase triggers elongation while MEL 11/myosin phosphatase targeting subunit inhibits this contractile event. We describe mutations in the nonmuscle myosin heavy chain gene nmy-1 that were isolated as suppressors of the mel-11 hypercontraction phenotype. However, a nmy 1 null allele displays elongation defects less severe than mutations in let-502 or in the single nonmuscle myosin light chain gene mlc-4. This results because nmy-1 is partially redundant with another nonmuscle myosin heavy chain, nmy-2, which was previously known only for its role in anterior/posterior polarity and cytokinesis in the early embryo. At the onset of elongation, NMY-1 forms filamentous-like structures similar to actin, and LET-502 is interspersed with these structures, where it may trigger contraction. MEL-11, which inhibits elongation, is initially cytoplasmic. In response to LET-502 activity, MEL-11 becomes sequestered away from the contractile apparatus, to the plasma membrane, when elongation commences. Upon completion of morphogenesis, MEL-11 again appears in the cytoplasm where it may halt actin/myosin contraction. PMID- 14522876 TI - Neural crest development is regulated by the transcription factor Sox9. AB - The neural crest is a transient migratory population of stem cells derived from the dorsal neural folds at the border between neural and non-neural ectoderm. Following induction, prospective neural crest cells are segregated within the neuroepithelium and then delaminate from the neural tube and migrate into the periphery, where they generate multiple differentiated cell types. The intrinsic determinants that direct this process are not well defined. Group E Sox genes (Sox8, Sox9 and Sox10) are expressed in the prospective neural crest and Sox9 expression precedes expression of premigratory neural crest markers. Here, we show that group E Sox genes act at two distinct steps in neural crest differentiation. Forced expression of Sox9 promotes neural-crest-like properties in neural tube progenitors at the expense of central nervous system neuronal differentiation. Subsequently, in migratory neural crest cells, SoxE gene expression biases cells towards glial cell and melanocyte fate, and away from neuronal lineages. Although SoxE genes are sufficient to initiate neural crest development they do not efficiently induce the delamination of ectopic neural crest cells from the neural tube consistent with the idea that this event is independently controlled. Together, these data identify a role for group E Sox genes in the initiation of neural crest development and later SoxE genes influence the differentiation pathway adopted by migrating neural crest cells. PMID- 14522877 TI - The RNA-binding protein Vg1 RBP is required for cell migration during early neural development. AB - After mid-blastula transition, populations of cells within the Xenopus embryo become motile. Using antisense morpholino oligonucleotides, we find that Vg1 RBP, an RNA-binding protein implicated in RNA localization in oocytes, is required for the migration of cells forming the roof plate of the neural tube and, subsequently, for neural crest migration. These cells are properly determined but remain at their site of origin. Consistent with a possible role in cell movement, Vg1 RBP asymmetrically localizes to extended processes in migrating neural crest cells. Given that Vg1 RBP is a member of the conserved VICKZ family of proteins, expressed in embryonic and neoplastic cells, these data shed light on the likely role of these RNA-binding proteins in regulating cell movements during both development and metastasis. PMID- 14522878 TI - Segment boundary formation in Drosophila embryos. AB - In Drosophila embryos, segment boundaries form at the posterior edge of each stripe of engrailed expression. We have used an HRP-CD2 transgene to follow by transmission electron microscopy the cell shape changes that accompany boundary formation. The first change is a loosening of cell contact at the apical side of cells on either side of the incipient boundary. Then, the engrailed-expressing cells flanking the boundary undergo apical constriction, move inwards and adopt a bottle morphology. Eventually, grooves regress, first on the ventral side, then laterally. We noted that groove formation and regression are contemporaneous with germ band retraction and shortening, respectively, suggesting that these rearrangements could also contribute to groove morphology. The cellular changes accompanying groove formation require that Hedgehog signalling be activated, and, as a result, a target of Ci expressed, at the posterior of each boundary (obvious targets like stripe and rhomboid appear not to be involved). In addition, Engrailed must be expressed at the anterior side of each boundary, even if Hedgehog signalling is artificially maintained. Thus, there are distinct genetic requirements on either side of the boundary. In addition, Wingless signalling at the anterior of the domains of engrailed (and hedgehog) expression represses groove formation and thus ensures that segment boundaries form only at the posterior. PMID- 14522879 TI - Frontiers in cancer prevention research. PMID- 14522880 TI - Farnesyltransferase inhibitors: an overview of the results of preclinical and clinical investigations. AB - This article presents an overview of preclinical studies and clinical trials of a number of independently derived farnesyltransferase inhibitors (FTIs). Potential targets and biological modes of action of FTIs are discussed, and the results of clinical trials are summarized. The significant efficacy of FTIs as single or combined agents in preclinical studies stands in contrast with only moderate effects in clinical Phase II-III trials. These results reveal a substantial gap in the understanding of the complex activity of FTIs and their interactions with cytotoxic agents. We conclude that the rational combination of FTIs with other therapies, taking into account the biological activities of the individual agents, may improve the clinical results obtained with FTIs. PMID- 14522881 TI - Treatment of metastatic melanoma with an orally available inhibitor of the Ras Raf-MAPK cascade. AB - The Ras-Raf-MAPK pathway is constitutively activated in the majority of melanomas because of a mutation in the BRAF gene. It has been hypothesized that activation of this pathway is crucial for the genesis and maintenance of melanoma and therefore represents an attractive clinical target for metastatic disease. We synthesized a previously characterized MAP kinase kinase inhibitor to test the effect that blocking the Ras-Raf-MAPK pathway would have on the establishment and maintenance of melanoma metastases. Oral administration of CI 1040 inhibited formation of pulmonary metastases and caused rapid regression of established pulmonary metastases in the mouse. Our findings indicate that Ras-Raf-MAPK activation provides crucial signals for the survival of melanoma cells at ectopic sites and that the pharmacological inhibition of this pathway is a promising target for melanoma therapy. PMID- 14522882 TI - Deregulation of the cyclin D1/Cdk4 retinoblastoma pathway in rat mammary gland carcinomas induced by the food-derived carcinogen 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine. AB - 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is a suspected human breast carcinogen found in cooked meat that induces mammary gland cancer in rats. By real time PCR analysis, PhIP-induced rat mammary gland carcinomas showed statistically higher expression of the G(1)-S cyclin D1 (5-fold) and its kinase partner cyclin-dependent kinase (Cdk)-4 (37-fold) in comparison with normal mammary gland, whereas cyclin D2, cyclin D3, and Cdk6 were not statistically changed. Amplification of cyclin D1 was observed by real time PCR in 24% of carcinomas (15 of 63). Only 1 of 47 carcinomas showed Cdk4 amplification. By Western blotting, the level of phospho-Rb was >2-fold higher in carcinomas than in normal mammary gland. By immunohistochemical analysis, cyclin D1, Cdk4, and phospho-Rb nuclear protein expression was 5.7-, 3.9-, and 2.3-fold higher, respectively, in carcinomas than in normal mammary gland, whereas the expression of cyclin D2, cyclin D3, and Cdk6 was similar. Among carcinomas, Cdk4 and phospho Rb levels were positively correlated with cell proliferation. Previous studies by this laboratory indicated that these carcinomas harbor a high frequency of H-ras mutations. The H-ras pathway is linked to the cell cycle via cyclin D1. The results from the current study implicate cyclin D1/Cdk4, phospho-Rb as a central pathway in PhIP-induced rat mammary gland carcinogenesis. PMID- 14522883 TI - Molecular signature associated with bone marrow micrometastasis in human breast cancer. AB - Metastasis is the leading cause of cancer-related death, and bone marrow (BM) is a prominent metastatic site in solid tumors. Here, we focused on the onset of metastasis, using BM as an indicator organ for micrometastatic tumor cells in breast cancer patients without overt metastases (tumor-node-metastasis stage M(0)). Expression analysis with cDNA arrays showed distinct profiles between primary tumors from BM-positive and BM-negative patients. The differentially expressed genes are involved in extracellular matrix remodeling, adhesion, cytoskeleton plasticity, and signal transduction (in particular RAS and hypoxia inducible factor 1alpha pathway). The BM signature was mainly characterized by transcriptional repression and different from the expression signature associated with lymphatic metastasis. Thus, BM micrometastasis is a selective process with a specific molecular signature of the primary tumor. PMID- 14522884 TI - A peptide with three hyaluronan binding motifs inhibits tumor growth and induces apoptosis. AB - A number of hyaluronan (HA) binding proteins such as soluble CD44, receptor for hyaluronan-mediated motility (RHAMM), and metastatin inhibit tumor growth and metastasis. To determine whether the HA binding motif is the element responsible for the antitumor effect of this family of proteins, we examined the biological activity of a 42-amino acid peptide (designated as BH-P) that contains three HA binding motifs [B(X(7))B] from human brain HA binding protein. In initial experiments with cultured cells, we found that synthetic BH-P inhibited the proliferation and colony formation of tumor cells. It also blocked the growth of tumors on the chorioallantoic membranes of 10-day chicken embryos. In addition, MDA-435 melanoma cells that had been transfected with an expression vector for BH P grew at a slower rate in nude mice than the vector-alone transfectants. Final studies revealed that the BH-P could activate caspase-8, caspase-3, and poly(ADP ribose) polymerase and trigger the apoptosis of tumor cells. Taken together, these results suggest that the HA binding motif that is present in HA binding proteins may be responsible for the antitumor effect exerted by the members of this family. PMID- 14522885 TI - Identification of ZASC1 encoding a Kruppel-like zinc finger protein as a novel target for 3q26 amplification in esophageal squamous cell carcinomas. AB - Among the transcription factors that direct proliferation, differentiation, and death of cells, several Kruppel-like zinc finger molecules such as GLI1 and ZNF147 can become oncogenic when the genes encoding them are overexpressed by the DNA amplification mechanism. Here, we report the discovery of a novel member of this class, ZASC1, from a recently reported critical region of 3q26 amplification frequently observed in various squamous cell carcinomas, including esophageal squamous cell carcinomas (ESCs). The deduced 485-amino acid protein product of ZASC1 contained a putative nuclear localization signal; the exogenously transfected ZASC1 was translocated into cell nuclei. ZASC1 was frequently coamplified and subsequently overexpressed with PIK3CA in our panel of ESC cell lines and primary tumors. In patients with ESC, a higher mRNA expression level of ZASC1 appeared to be associated with shorter overall survival, and a multivariate analysis demonstrated that ZASC1 mRNA expression was an independent prognosticator. In addition, exogenous expression of ZASC1 promoted the growth of ESC cells, whereas down-regulation of ZASC1 expression by means of an antisense oligonucleotide suppressed the growth of ESC cells. Taken together, our results suggest that ZASC1 might be involved in the pathogenesis of ESC as one of the targets for 3q26 amplification, alone or in concert with other targets. PMID- 14522886 TI - Differential gene expression profile in endometrioid and nonendometrioid endometrial carcinoma: STK15 is frequently overexpressed and amplified in nonendometrioid carcinomas. AB - Endometrial carcinoma (EC) comprises at least two types of cancer: endometrioid carcinomas (EECs) are estrogen-related tumors, which are frequently euploid and have a good prognosis. Nonendometrioid carcinomas (NEECs; serous and clear cell forms) are not estrogen related, are frequently aneuploid, and are clinically aggressive. We used cDNA microarrays containing 6386 different genes to analyze gene expression profiles in 24 EECs and 11 NEECs to identify differentially expressed genes that could help us to understand differences in the biology and clinical outcome between histotypes. After supervised analysis of the microarray data, there was at least a 2-fold difference in expression between EEC and NEEC in 66 genes. The 31 genes up-regulated in EECs included genes known to be hormonally regulated during the menstrual cycle and to be important in endometrial homeostasis, such as MGB2, LTF, END1, and MMP11, supporting the notion that EEC is a hormone-related neoplasm. Conversely, of the 35 genes overexpressed in NEECs, three genes, STK15, BUB1, and CCNB2, are involved in the regulation of the mitotic spindle checkpoint. Because STK15 amplification/overexpression is associated with aneuploidy and an aggressive phenotype in other human tumors, we used fluorescence in situ hybridization to investigate whether STK15 amplification occurred in ECs. We found that STK15 was amplified in 55.5% of NEECs but not in any EECs (P 60% of cutaneous melanomas and premalignant melanocytic lesions. We have investigated the frequency of BRAF mutations at the expression level in melanomas of the uveal tract. None of the 30 metastases and 10 primary uveal melanomas tested expressed the V599E mutation. In contrast, this mutation was expressed by 65% of cutaneous melanoma samples, confirming previous results. In addition, a double mutation resulting in V599K substitution was detected in two suspect ocular metastases of cutaneous melanoma. Analysis of exon 11, the second common site of BRAF mutations, revealed only wild type sequences in uveal melanomas. Analysis of tumor lysates showed the presence of phosphorylated mitogen-activated protein kinase, kinase, and mitogen-activated protein kinase in 50% of uveal and 100% of cutaneous melanoma metastases. Taken together, these results suggest that although the common BRAF mutations found in cutaneous melanoma do not play a role in tumorigenesis of uveal tract melanocytes, activation of the RAF/mitogen-activated protein kinase pathway may nevertheless play an important role in uveal melanoma. PMID- 14522890 TI - Bcr-Abl kinase modulates the translation regulators ribosomal protein S6 and 4E BP1 in chronic myelogenous leukemia cells via the mammalian target of rapamycin. AB - Identification of signaling pathways downstream of Abl tyrosine kinase may increase our understanding of the pathogenesis of chronic myelogenous leukemia (CML) and suggest strategies to improve clinical treatment of the disease. By combining the use of a phosphospecific antibody recognizing a substrate motif of serine/threonine kinases with bioinformatics, we found that the translational regulators ribosomal protein S6 and 4E-BP1 are constitutively phosphorylated in CML cells. Experiments with specific inhibitors indicated the phosphorylation is downstream of Bcr-Abl kinase and the mammalian target of rapamycin (mTOR). These results suggest that Bcr-Abl may regulate translation of critical targets in CML cells via mTOR. They also provide a rationale for testing the combination of mTOR inhibitors with the Abl kinase inhibitor imatinib in patients with CML. The mTOR inhibitor rapamycin enhanced imatinib-mediated killing of CML cell lines in vitro, and it overcame imatinib resistance in cells with Bcr-Abl gene amplification. PMID- 14522891 TI - High-throughput molecular analysis of urine sediment for the detection of bladder cancer by high-density single-nucleotide polymorphism array. AB - The detection of urothelial malignancies remains challenging. The majority of patients diagnosed with bladder cancer require life-long surveillance for disease recurrence. Monitoring strategies rely predominantly on invasive endoscopic techniques, which are inconvenient and uncomfortable. Multiple in vitro diagnostic technologies have been developed to supplant the contemporary standard of care. The U.S. Food and Drug Administration has approved several assays, but [because of inferior performance characteristics (low sensitivity and specificity)] these tests have not made a significant impact on practice, to date. We sought to develop a test for bladder cancer with better performance characterization detection based on a novel molecular approach. Matched urine and peripheral blood lymphocyte samples were obtained before surgery from 31 patients with bladder cancer (10 pTa, 4 pT1, and 17 pT2>or). DNA from these samples was subjected to allelic imbalance analysis using HuSNP chips and was validated in parallel with microsatellite analysis for loss of heterozygosity and microsatellite instability. Peripheral blood lymphocyte and urine DNA obtained from 14 individuals without clinical evidence of genitourinary malignancy served as controls. Thirty-one of 31 (100%) urine DNA samples from patients with bladder tumors were found to have 24 or more single-nucleotide polymorphism (SNP) DNA alterations. In general, SNP alterations were more common in urine samples from pT2>or tumors than pTa or pT1 tumors. SNP alterations were not identified in nine normal control subjects and in four of five patients with hematuria. These data support the noninvasive HuSNP chip assay in urine DNA as a valuable tool for the detection of bladder cancer (on a high-throughput-automated platform). PMID- 14522892 TI - Association of breast cancer risk with a GT dinucleotide repeat polymorphism upstream of the estrogen receptor-alpha gene. AB - Recent studies suggest that genetic polymorphisms of the estrogen receptor-alpha (ER-alpha) gene may be associated with breast cancer risk. To evaluate the role of this gene in the risk of breast cancer, we genotyped a newly identified GT dinucleotide repeat [(GT)(n)] polymorphism located in the promoter region (6.6 kb upstream of the transcription start site) in 947 breast cancer cases and 993 age frequency-matched community controls from a population-based case-control study conducted among Chinese in urban Shanghai. Sixteen alleles were identified, the most common one having 16 GT repeats [(GT)(16)]. Compared with subjects homozygous for this allele, subjects carrying the (GT)(17) or (GT)(18) allele had a decreased risk of breast cancer. The odds ratios (ORs) were 0.81 [95% confidence interval (CI), 0.62-1.06] and 0.58 (95% CI, 0.36-0.94), respectively, for one and two copies of the (GT)(17) or (GT)(18) allele. The inverse association with carrying either of these alleles was stronger among women with >30 years of menstrual cycles (OR 0.66; 95% CI 0.51-0.85) than those with a shorter duration of menstrual cycles (OR 0.97; 95% CI 0.73-1.27), and the test for an interaction was statistically significant (P = 0.04). Among breast cancer patients, the presence of either the (GT)(17) or (GT)(18) allele was associated with a reduced expression of progesterone receptor. Results of this study indicate that the GT dinucleotide repeat polymorphism in ER-alpha gene promoter region may be a new biomarker for genetic susceptibility to breast cancer. PMID- 14522893 TI - Common genetic evolutionary pathways in familial adenomatous polyposis tumors. AB - Cancer cells progress through the accumulation of genetic alterations. Familial adenomatous polyposis (FAP) tumors provide an excellent model to unravel the molecular steps underlying malignant transformation. Global genomic damage was assessed in 56 adenomas and 3 carcinomas from six FAP patients and compared with that of sporadic adenomas and carcinomas. Evolutive trees were traced after application of maximum likelihood clustering and split decomposition methods to the analysis of comprehensive genetic profiles generated by diverse molecular approaches: arbitrarily primed PCR, comparative genomic hybridization, and flow cytometry. Overall, genomic damage as assessed by arbitrarily primed PCR was lower in familial adenomas than in sporadic adenomas and carcinomas. Comparative genomic hybridization data also show a low number of alterations in the majority of FAP adenomas. Tumors of the same patient were likely to share specific genetic alterations and may be grouped into one or two clusters. Putative common pathways were also identified, which included tumors of up to three different patients. According to our data, FAP tumors accumulate specific genetic alterations and in a preferred order that is characteristic of each individual. Moreover, the particular genetic background and environmental conditions of a FAP patient restrain the molecular evolution portrait of synchronous tumors. PMID- 14522894 TI - Microsatellite instability is a predictive factor of the tumor response to irinotecan in patients with advanced colorectal cancer. AB - The aim of our study was to assess the relationship between colorectal tumor responsiveness to irinotecan and microsatellite instability (MSI), a feature of colorectal tumors with DNA mismatch repair defect. Seventy-two patients with metastatic colorectal cancer were included in our retrospective study. A complete response to irinotecan was observed in 1 patient and a partial response in 10 patients, whereas 61 patients did not respond to this treatment. We analyzed the protein expression of hMLH1, hMSH2, and BAX by immunohistochemistry, determined the MSI phenotype, and looked for mutations in the coding repeats located in the transforming growth factor beta-RII, BAX, hMSH3, and hMSH6 genes. All 44 tumors analyzed expressed detectable levels of hMLH1; 1 tumor lacked hMSH2 staining, whereas 4 tumors showed a marked decrease in BAX expression. A better response to irinotecan was observed in the patients whose tumors have lost BAX expression (P < 0.001). Among the 7 tumors that displayed a MSI-H phenotype, 4 responded to irinotecan, whereas only 7 of the 65 MSI-L/ microsatellite stable tumors did (P = 0.009). Seven of the 72 tumors had inactivating mutations in the coding repeats of the target genes. Three tumors displayed a mutation in the poly-A10 tract of the transforming growth factor beta-RII gene, associated with a 1-bp deletion in the poly-A8 tract of hMSH3 in one tumor and with a 1-bp deletion in the poly-G8 tract of BAX in another. Four tumors displayed mutations in the poly-G8 repeat of BAX, whereas 2 mutations in hMSH6 and hMSH3 were characterized. Among the 7 tumors with mutations in these target genes, 5 responded to irinotecan, whereas only 6 of the other 65 tumors did (P < 0.001), indicating that MSI-driven inactivation of target genes modifies tumor chemosensitivity. Our observations allowed us to define the first useful predictive criteria for irinotecan response in patients with colorectal cancer. PMID- 14522895 TI - Inhibition of induced chemoresistance by cotreatment with (E)-5-(2-bromovinyl)-2' deoxyuridine (RP101). AB - Induced chemoresistance leads to the reduction of apoptotic responses. Although several drugs are in development that circumvent or decrease existing chemoresistance, none has the potential to prevent or reduce its induction. Here, we present data from a drug that could perhaps fill this gap. Cotreatment of chemotherapy with (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU, RP101) prevented the decrease of apoptotic effects during the course of chemotherapy and reduced nonspecific toxicity. Amplification of chemoresistance genes (Mdr1 and Dhfr) and overexpression of gene products involved in proliferation (DDX1) or DNA repair (UBE2N and APEX) were inhibited, whereas activity of NAD(P)H: quinone oxidoreductase 1 (NQO1) was enhanced. During recovery, when treatment was with BVDU only, microfilamental proteins were up-regulated, and proteins involved in ATP generation or cell survival (STAT3 and JUN-D) were down-regulated. That way, in three different rat tumor models, the antitumor efficiency of chemotherapy was optimized, and toxic side effects were reduced. Because of these beneficial properties of BVDU, a clinical pilot Phase I/II study with five human tumor entities has been started at the University of Dresden (Dresden, Germany). So far, no unwanted side effects have been observed. PMID- 14522896 TI - Fibroblast growth factor 2 promotes tumor progression in an autochthonous mouse model of prostate cancer. AB - Fibroblast growth factor (FGF) 2 (or basic FGF) is expressed at increased levels in human prostate cancer. FGF2 can promote cell motility and proliferation, increase tumor angiogenesis, and inhibit apoptosis, all of which play an important role in tumor progression. To determine whether FGF2 plays a critical role in prostate cancer progression, we have used the transgenic adenocarcinoma of the mouse prostate (TRAMP) model system. A high percentage of TRAMP mice develop metastatic prostate cancer, and thus the TRAMP model is useful for evaluating cancer progression. TRAMP mice were crossed with FGF2 knockout (FGF2( /-)) mice, and tumor progression in TRAMP mice that were either hemi- or homozygous for inactivation of the FGF2 allele was compared with progression in wild-type TRAMP mice. Inactivation of even one FGF2 allele resulted in increased survival, a decrease in metastasis, and inhibition of progression to the poorly differentiated phenotype in primary prostatic tumors. When compared with wild type mice, poorly differentiated tumors arising in FGF(+/-) and FGF(-/-) mice expressed higher levels of vascular endothelial growth factor and, in some cases, increased levels of acidic FGF intracellular binding protein, a nuclear FGF1 binding protein. These findings suggest that both FGF2-mediated angiogenesis and intranuclear FGF2 activities may promote tumor progression and support the hypothesis that FGF2 plays a significant role in prostate cancer progression in vivo. PMID- 14522897 TI - Absence of BRAF and NRAS mutations in uveal melanoma. AB - Uveal melanoma (UM) and cutaneous melanoma (CM) differ significantly in their epidemiological, clinical, immunophenotypical, and cytogenetic features, but the molecular basis for these differences has not been delineated. CMs frequently harbor an activating mutation in either NRAS or the RAS-regulated kinase BRAF, suggesting that either of these oncogenes may increase signaling through the mitogen-activated protein (MAP) kinase pathway and promote melanoma development. The aim of this study was to examine BRAF and NRAS gene mutations in UM. Genomic DNA from CM and UM was screened for mutations in BRAF exons 11 and 15 and NRAS exons 1 and 2 using a combination of denaturing high-performance liquid chromatography and direct sequencing. Mutations in BRAF exon 15 were detected in 16 (36.4%) of 44 CMs and 0 (0%) of 62 UMs. The most common mutation in CM was V599E, but a novel point mutation (L596Q) was identified in two cases and an in frame deletion/insertion (VKSRWK599-604D) was discovered in one case. No BRAF exon 11 mutations were observed among seven CMs and nine UMs that were wild-type for exon 15. Mutation of NRAS exon 2 was rare in CM [1 (3.7%) of 27] and absent in UM [0 (0%) of 47]. No NRAS exon 1 mutations were detected in either type of melanoma. We conclude that UMs arise independent of oncogenic BRAF and NRAS mutations, an observation that may have implications for therapies targeted to the NRAS-BRAF pathway. PMID- 14522898 TI - Comprehensive gene expression analysis of peroxisome proliferator-treated immortalized hepatocytes: identification of peroxisome proliferator-activated receptor alpha-dependent growth regulatory genes. AB - Chemicals known as peroxisome proliferators (PPs) are the subject of intense study because of their ability to cause hepatocellular carcinoma in laboratory rodents. These chemicals act through a family of proteins termed the peroxisome proliferator-activated receptors (PPARs), in particular PPARalpha. It has become increasingly apparent that the role of the PPs in the development of cancer encompasses many different aspects of cell growth regulation. Immortalized hepatocytes from wild-type (PPARalpha(+/+)) and PPARalpha(-/-) mice were generated using a temperature-sensitive SV40 virus. Characterization of the murine SV40 hepatocytes (MuSH) generated from both genotypes (MuSHalpha(+/+), MuSHalpha(-/-)) show markers of differentiation such as albumin expression, but is devoid of Kupffer cell contamination. Hallmark PPARalpha-mediated responses such as induction of acyl-CoA oxidase mRNA by PPs are present in the MuSHalpha(+/+) but are absent in MuSHalpha(-/-) cells. In contrast to most cell culture systems, the wild-type MuSH hepatocytes retain the mitogenic activity of PPs, whereas the MuSHalpha(-/-) does not respond in this manner, thus making this cell culture system an ideal tool to examine growth regulatory gene expression affected by PPs. Microarray experiments performed on both cell types identified many genes in which regulation is dependent on the presence of PPARalpha, and these changes were verified with reverse transcriptase-PCR. Genes involved in carcinogenesis and control of the cell cycle that are regulated by PPs in a PPARalpha-dependent manner include ubiquitin COOH-terminal hydrolase 37 (also known as UCT-L5) and cyclin T1. These results show that MuSH cells reflect the biological properties of both the wild-type and PPARalpha-null animals and can be used to identify novel PPARalpha-regulated genes that could be involved in regulation of the cell cycle and carcinogenesis. PMID- 14522899 TI - Genome-wide identification of chromosomal regions of increased tumor expression by transcriptome analysis. AB - Genes up-regulated in tumor cells provide attractive anticancer therapeutic targets. Although the general underlying mechanism for the increased expression in tumors is unknown, tumor-specific up-regulation of some genes can be attributed to aberrant DNA amplification, a phenomenon common to many tumors. Using a computational method, we constructed a general transcriptome map with the human genomic sequences and expressed sequence tags in the public database. The transcriptome map revealed nonrandom chromosomal regions (termed region of increased tumor expression) where clusters of genes exhibited increased expression in the 10 tumor tissue types tested. These genomic regions often correspond to experimentally verified tumor amplicons. Our large-scale transcriptome analysis led to identification of many additional chromosomal regions with increased tumor expression, regions that represent potential tumor amplicons. PMID- 14522900 TI - ING1 represses transcription by direct DNA binding and through effects on p53. AB - The ING family of proteins is involved in the regulation of diverse processes ranging from cell cycle and cellular senescence to apoptosis. These effects are most likely through activation of acetylation-dependent pathways that ultimately alter gene expression. Despite reports linking ING to p53 activation, the molecular basis of how ING activates p53 function has not been elucidated. In this study, we found that a subset of ING family members strongly repressed human alpha-fetoprotein (AFP) promoter activity but stimulated the p21(WAF1) promoter in parallel experiments in the same cell type, similar to the effects of p53. The p47(ING1a) isoform also repressed AFP promoter activity, but in contrast to other ING isoforms, it repressed the p21(WAF1) promoter. p47(ING3) up-regulated p21(WAF1) promoter activity, but it did not have any effect on the AFP promoter. ING1b and ING2 also repressed the AFP promoter in Hep3B p53-null cell lines, and p53 coexpression enhanced this transcriptional repression. Suppression of AFP gene transcription by ING was strongly dependent on AT-motifs that bind to the hepatocyte nuclear factor 1 (HNF1) transcription factor. Indeed, electrophoretic mobility shift assays confirmed that HNF1 binds to AT-motifs, but we found, surprisingly, that the ING1 complexes binding to these AT-motifs were devoid of HNF1 protein. Both ING1 and p53 were able to suppress AFP transcription and cause p21 induction; hSIR2, a negative regulator of the p53 protein, showed the opposite effects on the AFP promoter and, like HDAC1, repressed p21 promoter activity. In addition, we found that p33(ING1b) physically interacts with hSIR2, reverses its ability to induce the AFP promoter, and induces acetylation of p53 residues at Lys(373) and/or Lys(382). These findings provide novel evidence that p33(ING1b) represses AFP transcription by at least two mechanisms, one of which includes p53. The first is by binding to the AT-motif and excluding HNF1 binding while possibly targeting HAT activity to promoter regions, and the second is by increasing the levels of active, acetylated p53 via binding and inhibiting the ability of hSIR2 to deacetylate p53 protein. PMID- 14522901 TI - The mutational spectrum of the HPRT gene from human T cells in vivo shares a significant concordant set of hot spots with MNNG-treated human cells. AB - The preponderance of G:C to A:T transitions in inherited and somatic human mutations has led to the hypothesis that some of these mutations arise as a result of formation of O(6)-methylguanine in DNA. To test this hypothesis, the fine structure map of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced mutations was determined in human lymphoblastoid cells in the human hypoxanthine guanine-phosphoribosyltransferase (HPRT) gene and compared with HPRT mutations observed in somatic T lymphocytes from normal individuals. Human TK6 cells, which are methylguanine methyltransferase deficient (MGMT(-)), were treated with the methylating agent MNNG to create a level of O(6)-methylguanine in cellular DNA equal to that found in normal human tissues. A total of 676 bp of the HPRT gene was scanned using constant denaturing capillary electrophoresis and high-fidelity PCR. MNNG induced 14 predominant hot spots, all which were G:C to A:T transitions. Thirteen of these 14 MNNG-induced hot spots were found among the in vivo set, and 10 of the MNNG-induced hot spots were among 75 putative in vivo hot spots (mutations observed two or more times in vivo). Using a hypergeometric test for concordance, the MNNG-induced hot spots were found to be a significant subset of the putative in vivo hot spots (P < 4 x 10(-7)). The set of shared hot spots comprise some 18% of the HPRT in vivo hot spot spectrum and strongly suggest that MNNG-induced hot spots in vitro share a common mutational pathway with a significant subset of somatic mutations in vivo. PMID- 14522903 TI - Congenic rats reveal three independent Copenhagen alleles within the Mcs1 quantitative trait locus that confer resistance to mammary cancer. AB - It has previously been shown that the Copenhagen (COP) rat contains several genetic loci that contribute to its mammary tumor-resistant phenotype after 7,12 dimethylbenz(a)anthracene (DMBA) administration. One of these loci, mammary carcinoma susceptibility 1 (Mcs1), is located on the centromeric end of chromosome 2 and appears to act in a semidominant fashion. To confirm the existence and independent action of this locus and also aid in the identification of the physical location of the Mcs1 gene, congenic lines were generated by transferring the Mcs1 COP allele onto a Wistar Furth (WF) genetic background. Male carriers were genotyped using microsatellite markers spanning 20-30 cM of the Mcs1 locus. One of the congenic lines minimally retained the COP allele at D2Mit29 on the centromeric end of chromosome 2 and extended distally to D2Rat201. Heterozygous Mcs1 carrier rats were interbred, and the female offspring were treated with DMBA. The female rats from the Mcs1 congenic line that carried one or two COP alleles of the Mcs1 region had a significantly reduced (65 and 85%, respectively) tumor development (P < 0.001) compared with rats carrying zero COP alleles at this locus. A WF.COP-D2Mit29/D2Rat201 homozygous congenic strain derived at the N10 generation was treated with DMBA, and the COP homozygous rats developed 1.5 +/- 0.3 carcinomas/rat versus 6.3 +/- 0.5 in WF control rats (P < 0.0001). Fine mapping of this congenic interval using several recombinant lines identified three genetic loci within the Mcs1 congenic region that independently supported a tumor resistance phenotype. These genetic loci have been termed Mcs1a, Mcs1b, and Mcs1c. In rats for which each locus was homozygous for the COP allele, tumor development was reduced by approximately 60% compared with littermate controls. The identification of these independent loci within the Mcs1 COP allele provide a model of the genetic complexity of cancer. PMID- 14522902 TI - Base excision repair deficiency caused by polymerase beta haploinsufficiency: accelerated DNA damage and increased mutational response to carcinogens. AB - The base excision repair pathway (BER) is believed to maintain genomic integrity by repairing DNA damage arising spontaneously or induced by oxidizing and alkylating agents. To establish the role of DNA polymerase beta (beta-pol) in BER and beta-pol-dependent BER in maintaining genomic stability, we have measured the impact of a gene-targeted disruption in the beta-pol gene on DNA repair capacity and on in vivo sensitivity to carcinogens. We have extensively phenotyped the DNA beta-pol heterozygous (beta-pol(+/-)) mouse as expressing approximately 50% less beta-pol mRNA and protein and as exhibiting an equivalent reduction in the specific activity of beta-pol. We measured BER activity by in vitro G:U mismatch and (8-OH)G:C repair and find that there is a significant reduction in the ability of extracts from beta-pol(+/-) mice to repair these types of DNA damage. In vivo, the beta-pol(+/-) mice sustain higher levels of DNA single-strand breaks as well as increased chromosomal aberrations as compared with beta-pol(+/+) littermates. Additionally, we show that reduction in beta-pol expression and BER activity results in increased mutagenicity of dimethyl sulfate as evidenced by a 2-fold increase in LacI mutation frequency. Importantly, the beta-pol(+/-) mice do not exhibit increased sensitivity to DNA damage induced by N-nitroso-N ethylurea, ionizing radiation, or UV radiation, which induce damage processed by alternative repair pathways, demonstrating that this model is specifically a BER deficient model. PMID- 14522904 TI - Aneuploidy and telomere attrition are independent determinants of crisis in SV40 transformed epithelial cells. AB - Replicative immortality is achieved in vitro by overcoming two mortality checkpoints, M1 (senescence) and M2 (crisis). Cancer cells are thought to overcome M2 by activating telomerase, an enzyme believed to confer genomic stability in addition to maintaining telomeric sequences above a critical length. Here we show that a subset of cultured ovarian cystadenoma cells expressing SV40 large T-antigen, which allows bypassing of M1, develop a specific type of genomic instability, characterized by numerical (as opposed to structural) chromosomal alterations, that leads to non-telomere-based premature growth arrest/crisis. Cells recover from this type of growth arrest and stabilize their ploidy status without telomerase expression. In these cases, telomeres continue to shorten until a second, telomere-based growth arrest/crisis event is reached. Transfection of the catalytic subunit of telomerase does not immortalize cells harboring severe abnormalities in their DNA ploidy but results in immortalization of diploid cells. We conclude that changes in DNA ploidy constitute an important determinant of growth arrest that is independent of telomere attrition in a subset of SV40 large T-antigen-expressing cystadenoma cells. Reestablishment or emergence of ploidy stability, which is not always dependent on telomerase activation, is necessary for acquisition of the potential to achieve replicative immortality. PMID- 14522905 TI - Identification of a cancer stem cell in human brain tumors. AB - Most current research on human brain tumors is focused on the molecular and cellular analysis of the bulk tumor mass. However, there is overwhelming evidence in some malignancies that the tumor clone is heterogeneous with respect to proliferation and differentiation. In human leukemia, the tumor clone is organized as a hierarchy that originates from rare leukemic stem cells that possess extensive proliferative and self-renewal potential, and are responsible for maintaining the tumor clone. We report here the identification and purification of a cancer stem cell from human brain tumors of different phenotypes that possesses a marked capacity for proliferation, self-renewal, and differentiation. The increased self-renewal capacity of the brain tumor stem cell (BTSC) was highest from the most aggressive clinical samples of medulloblastoma compared with low-grade gliomas. The BTSC was exclusively isolated with the cell fraction expressing the neural stem cell surface marker CD133. These CD133+ cells could differentiate in culture into tumor cells that phenotypically resembled the tumor from the patient. The identification of a BTSC provides a powerful tool to investigate the tumorigenic process in the central nervous system and to develop therapies targeted to the BTSC. PMID- 14522906 TI - p73 tumor-suppressor activity is impaired in human thyroid cancer. AB - The p73 protein is a member of the p53 family and, like p53, can induce cell cycle arrest and apoptosis in response to DNA damage. Because the loss of p53 function is responsible for the progression of well-differentiated thyroid cancer to more aggressive phenotypes, we hypothesized that p73 might also be involved in thyroid carcinogenesis. We find that normal thyrocites do not express p73, whereas most thyroid malignancies are positive for p73 expression. However, the p73 protein of thyroid cancer cells is unresponsive to DNA-damaging agents, failing to elicit a block of the cell cycle or an apoptotic response. Notably, overexpression of transcriptionally active p73 in thyroid cancer lines can arrest the cell cycle but is still unable to induce cell death. The loss of p73 biological activity in neoplastic thyroid cells is partly explained by its interaction with transcriptionally inactive variants of p73 (DeltaNp73) and with mutant p53. Our findings suggest that the functional impairment of p73 could be involved in the development of thyroid malignancies, defining p73 as a potential therapeutic target for thyroid cancer. PMID- 14522907 TI - Molecular imaging of angiogenesis in nascent Vx-2 rabbit tumors using a novel alpha(nu)beta3-targeted nanoparticle and 1.5 tesla magnetic resonance imaging. AB - Early noninvasive detection and characterization of solid tumors and their supporting neovasculature is a fundamental prerequisite for effective therapeutic intervention, particularly antiangiogenic treatment regimens. Emerging molecular imaging techniques now allow recognition of early biochemical, physiological, and anatomical changes before manifestation of gross pathological changes. Although new tumor, vascular, extracellular matrix, and lymphatic biomarkers continue to be discovered, the alpha(nu)beta(3)-integrin remains an attractive biochemical epitope that is highly expressed on activated neovascular endothelial cells and essentially absent on mature quiescent cells. In this study, we report the first in vivo use of a magnetic resonance (MR) molecular imaging nanoparticle to sensitively detect and spatially characterize neovascularity induced by implantation of the rabbit Vx-2 tumor using a common clinical field strength (1.5T). New Zealand White rabbits (2 kg) 12 days after implantation of fresh Vx-2 tumors (2 x 2 x 2 mm(3)) were randomized into one of three treatment groups: (a) alpha(nu)beta(3)-targeted, paramagnetic formulation; (b) nontargeted, paramagnetic formulation; and (c) alpha(nu)beta(3)-targeted nonparamagnetic nanoparticles followed by (2 h) the alpha(nu)beta(3)-targeted, paramagnetic formulation to competitively block magnetic resonance imaging (MRI) signal enhancement. After i.v. systemic injection (0.5 ml of nanoparticles/kg), dynamic T(1)-weighted MRI was used to spatially and temporally determine nanoparticle deposition in the tumor and adjacent tissues, including skeletal muscle. At 2-h postinjection, alpha(nu)beta(3)-targeted paramagnetic nanoparticles increased MRI signal by 126% in asymmetrically distributed regions primarily in the periphery of the tumor. Similar increases in MR contrast were also observed within the walls of some vessels proximate to the tumor. Despite their relatively large size, nanoparticles penetrated into the leaky tumor neovasculature but did not appreciably migrate into the interstitium, leading to a 56% increase in MR signal at 2 h. Pretargeting of the alpha(nu)beta(3)-integrin with nonparamagnetic nanoparticles competitively blocked the specific binding of alpha(nu)beta(3) targeted paramagnetic nanoparticles, decreasing the MR signal enhancement (50%) to a level attributable to local extravasation. The MR signal of adjacent hindlimb muscle or contralateral control tissues was unchanged by either the alpha(nu)beta(3)-targeted or control paramagnetic agents. Immunohistochemistry of alpha(nu)beta(3)-integrin corroborated the extent and asymmetric distribution of neovascularity observed by MRI. These studies demonstrate the potential of this targeted molecular imaging agent to detect and characterize (both biochemically and morphologically) early angiogenesis induced by minute solid tumors with a clinical 1.5 Tesla MRI scanner, facilitating the localization of nascent cancers or metastases, as well as providing tools to phenotypically categorize and segment patient populations for therapy and to longitudinally follow the effectiveness of antitumor treatment regimens. PMID- 14522908 TI - Dual stromelysin-3 function during natural mouse mammary tumor virus-ras tumor progression. AB - In human carcinomas, stromelysin-3/matrix metalloproteinase 11 (ST3, MMP11) expression by nonmalignant fibroblastic cells located in the immediate vicinity of cancer cells is a bad prognostic factor. Using mouse models of primary tumors, it has been demonstrated that ST3 is a key player during local invasion, favoring cancer cell survival in connective tissue through an antiapoptotic function. To investigate the ST3 impact on additional phases of cancer cell invasion, we developed mammary gland cancer prone MMTV-ras transgenic mice in wild-type (ras+/+;ST3+/+) or ST3-deficient (ras+/+;ST3-/-) genotype and studied their whole natural cancer history. The tumor-free survival and delay between the first ras oncogenic hit and primary tumor appearance increased in ras+/+;ST3-/- mice (P < 0.000001 and <0.0000007, respectively). A systematic search for occult primary tumors and metastases revealed, in addition to a lower total number and size of primary tumors (P < 0.02), an unexpected higher number of metastases (P < 0.01) in ras+/+;ST3-/- mice. Moreover, for a similar number and size of primary invasive tumors, ras+/+;ST3-/- mice developed more metastases, indicating that the cancer cells evolving in ST3-deficient stroma have an increased potential to hematogenous dissemination. We conclude that the ST3 microenvironment is a consistently active partner of invading cancer cells but that its function differs throughout cancer progression, being tumor enhancer or repressor in processes leading to local or distal invasion. Such a dual effect for an MMP might shed light, at least partially, for the absence of survival benefit for patients included in anti-MMP clinical trials. PMID- 14522909 TI - Insulin-like growth factor-1 induces adhesion and migration in human multiple myeloma cells via activation of beta1-integrin and phosphatidylinositol 3' kinase/AKT signaling. AB - Insulin-like growth factor-1 (IGF-I) is a growth and survival factor in human multiple myeloma (MM) cells. Here we examine the effect of IGF-I on MM cell adhesion and migration, and define the role of beta1 integrin in these processes. IGF-I increases adhesion of MM.1S and OPM6 MM cells to fibronectin (FN) in a time and dose-dependent manner, as a consequence of IGF-IR activation. Conversely, blocking anti-beta1 integrin monoclonal antibody, RGD peptide, and cytochalasin D inhibit IGF-I-induced cell adhesion to FN. IGF-I rapidly and transiently induces association of IGF-IR and beta1 integrin, with phosphorylation of IGF-IR, IRS-1, and p85(PI3-K). IGF-I also triggers phosphorylation of AKT and ERK significantly. Both IGF-IR and beta1 integrin colocalize to lipid rafts on the plasma membrane after IGF-I stimulation. In addition, IGF-I triggers polymerization of F-actin, induces phosphorylation of p125(FAK) and paxillin, and enhances beta1 integrin interaction with these focal adhesion proteins. Importantly, using pharmacological inhibitors of phosphatidylinositol 3'-kinase (PI3-K) (LY294002 and wortmannin) and extracellular signal-regulated kinase (PD98059), we demonstrate that IGF-I-induced MM cell adhesion to FN is achieved only when PI3 K/AKT is activated. IGF-I induces a 1.7-2.2 (MM.1S) and 2-2.5-fold (OPM6) increase in migration, whereas blocking anti-IGF-I and anti-beta1 integrin monoclonal antibodies, PI3-K inhibitors, as well as cytochalasin D abrogate IGF-I induced MM cell transmigration. Finally, IGF-I induces adhesion of CD138+ patient MM cells. Therefore, these studies suggest a role for IGF-I in trafficking and localization of MM cells in the bone marrow microenvironment. Moreover, they define the functional association of IGF-IR and beta1 integrin in mediating MM cell homing, providing the preclinical rationale for novel treatment strategies targeting IGF-I/IGF-IR in MM. PMID- 14522911 TI - Endothelial precursor cells as a model of tumor endothelium: characterization and comparison with mature endothelial cells. AB - Human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells (HMVEC) have been the standards for cell-based assays in the field of angiogenesis research and in antiangiogenic drug discovery. These normal mature endothelial cells may not be most representative of human tumor endothelial cells. Human AC133+/CD34+ bone marrow progenitor cells were established in cell culture media containing vascular endothelial growth factor, basic fibroblast growth factor (bFGF), and heparin to drive differentiation toward the endothelial phenotype. The resulting cells designated endothelial precursor cells (EPC) have many of the same functional properties as mature endothelial cells represented by HUVEC and HMVEC. By SAGE analysis, the genes expressed by EPC are more similar to the genes expressed by endothelial cells isolated from fresh surgical specimens of human tumors than are the genes expressed by HUVEC and HMVEC. Analysis of several cell surface markers by flow cytometry showed that EPC, HUVEC, and HMVEC have similar expression of P1H12, vascular endothelial growth factor 2, and endoglin but that EPC have much lower expression of ICAM1, ICAM2, VCAM1, and thrombomodulin than do HUVEC and HMVEC. The EPC generated can form tubes/networks on Matrigel, migrate through porous membranes, and invade through thin layers of Matrigel similarly to HUVEC and HMVEC. However, in a coculture assay using human SKOV3 ovarian cancer cell clusters in collagen as a stimulus for invasion through Matrigel, EPC were able to invade into the malignant cell cluster, whereas HMVEC were not able to invade the malignant cell cluster. In vivo, a Matrigel plug assay where human EPC were suspended in the Matrigel allowed tube/network formation by human EPC to be carried out in a murine host. EPC may be a better model of human tumor endothelial cells than HUVEC and HMVEC and, thus, may provide an improved cell based model for second generation antineoplastic antiangiogenic drug discovery. PMID- 14522910 TI - Promotion of angiogenesis by ps20 in the differential reactive stroma prostate cancer xenograft model. AB - Human prostate cancer is associated with a reactive stroma typified by an increase in the proportion of myofibroblast type cells and elevated synthesis of extracellular matrix proteins. Increased vascular density has been identified in the reactive stroma compartment adjacent to both precancerous and cancerous prostate lesions. The differential reactive stroma (DRS) prostate cancer xenograft model has been developed to investigate the role of reactive stroma in prostate cancer progression. Using this model, we have shown that human prostate stromal cells promote angiogenesis and growth of LNCaP human prostate carcinoma cell tumors, and that these increases are transforming growth factor (TGF) beta1 regulated. Our laboratory isolated and identified previously the ps20 protein (WFDC1 gene) as a prostate stromal cell secreted protein. The ps20 protein contains a whey acidic protein-type four-disulfide core domain, which is a functional motif characterized by serine protease inhibition activity in a number of whey acidic protein domain-containing proteins. In the present study, we show ps20 expression by normal human prostate stromal smooth muscle cells and vascular smooth muscle cells indicating a possible role of ps20 in vessel wall biology. Using in vitro assays, we show that ps20 promotes endothelial cell motility but has no effect on endothelial cell proliferation. To address the potential effects of ps20 in a tumor microenvironment, we used the DRS model to evaluate both angiogenesis and tumorigenesis of tumors generated under either ps20 or control conditions. DRS tumors generated with LNCaP and human prostate stromal cells in the presence of ps20 showed a 67% increase in microvessel density compared with control tumors. Elevated DRS tumor growth in the ps20-treated tumors was reflected by a 29% increase in wet weight and a 58% increase in volume compared with controls. Similar tumors composed of GeneSwitch-3T3 cells engineered to express ps20-V5-His under mifepristone regulation showed a 129% increase in microvessel density after induction of ps20-V5-His. GeneSwitch-3T3 cells expressing ps20-V5-His were localized to vessel walls in a mural cell (pericyte) position indicating a possible direct stabilizing interaction with endothelial cells. In addition, we show that ps20 mRNA synthesis is induced by TGF-beta1, a known regulator of endothelial cell-pericyte interactions and of stromal cell induced angiogenesis in DRS tumors. These findings suggest that ps20 may be a TGF beta1-induced regulator of angiogenesis that functions by either promoting endothelial cell migration or by contributing to pericyte stabilization of newly formed vascular structures. PMID- 14522912 TI - Early postoperative peripheral blood reverse transcription PCR assay for prostate specific antigen is associated with prostate cancer progression in patients undergoing radical prostatectomy. AB - Preoperative peripheral blood reverse transcription-PCR (RT-PCR) for prostate specific antigen (PSA) [RT-PCR-PSA] is not associated with an increased risk of progression after radical prostatectomy. We tested the hypothesis that early postoperative peripheral blood RT-PCR-PSA would detect prostate cancer cells persisting in the circulation that would be associated with disease progression. The study group consisted of 145 consecutive patients who underwent radical prostatectomy for clinically localized disease (median follow-up, 54.5 months) for whom pre- and postoperative peripheral blood samples were available. RT-PCR PSA was performed on preoperative and postoperative peripheral blood specimens. Pre- and postoperative RT-PCR-PSA were positive in 27% and 12%, respectively, of patients. Most (64%) preoperative RT-PCR-PSA-positive patients converted to a negative RT-PCR-PSA status after prostate removal (P < 0.001). Whereas preoperative RT-PCR-PSA was not associated with prostate cancer characteristics or outcome, a positive postoperative RT-PCR-PSA assay was associated with extracapsular extension (P = 0.044) and seminal vesicle involvement (P = 0.024). Furthermore, postoperative RT-PCR-PSA was an independent predictor of disease progression (P = 0.027). In patients who experienced disease progression, postoperative RT-PCR-PSA was associated with a more aggressive pattern of failure (P = 0.005). Whereas a significant number of patients with clinically localized prostate cancer have prostate cells detectable preoperatively by RT-PCR-PSA circulating in their blood, most of these cells are clinically insignificant because the majority of these patients convert to RT-PCR-PSA-negative status and maintain disease-free status after prostate removal. In contrast, postoperative RT-PCR-PSA detection of prostate cells in the peripheral blood is associated with established markers of aggressive prostate cancer and is an early independent predictor of disease progression, presumably because of an association with established micrometastatic disease. PMID- 14522913 TI - Aberrant HOXC expression accompanies the malignant phenotype in human prostate. AB - Dysregulation of HOX gene expression has been implicated as a factor in malignancies for a number of years. However, no consensus has emerged regarding specific causative genes. Using a degenerate reverse transcription-PCR technique, we show up-regulation of genes from the HOXC cluster in malignant prostate cell lines and lymph node metastases. When relative expression levels of the four HOX clusters were examined, lymph node metastases and cell lines derived from lymph node metastases exhibited very similar patterns, patterns distinct from those in benign cells or malignant cell lines derived from other tumor sites. Specific reverse transcription-PCR for HOXC4, HOXC5, HOXC6, and HOXC8 confirmed overexpression of these genes in malignant cell lines and lymph node metastases. Laser capture microdissection and examination of paired tumor/normal prostate epithelial cells also indicated overexpression of these HOXC genes in primary tumor cells. Our data indicate a possible link between expression of HOXC genes and malignancy in prostate cells. Overexpression of HOXC8 in LNCaP prostate cancer cells suppressed transactivation by androgen receptors. We speculate that HOXC overexpression may predispose tumor cells to androgen independence by necessitating adaptation to diminished androgen signaling. PMID- 14522914 TI - Serum levels of surfactant protein D are increased in mice with lung tumors. AB - Most murine lung tumors are composed of differentiated epithelial cells. We have reported previously that surfactant protein (SP)-D is expressed in urethane induced tumors. Serum levels of SP-D are increased in patients with interstitial lung disease and acute respiratory distress syndrome and in rats with acute lung injury but have not been measured in mice. In this study, we sought to determine whether SP-D could be detected in murine serum and discovered that it was increased in mice bearing lung tumors. Serum SP-D concentration was 5.0 +/- 0.2 ng/ml in normal C57BL/6 mice, essentially absent in SP-D nulls, and 63.6 +/- 9.0 ng/ml in SP-D-overexpressing mice. SP-D in serum was verified by immunoblotting. Serum SP-D was increased in mice bearing tumors induced by three different protocols, and the SP-D level correlated with tumor volume. However, in mice with a single adenoma or a few adenomas, SP-D levels were usually within the normal range. SP-D was expressed by the tumor cells, and there was also a field effect whereby type II cells near the tumor expressed more SP-D than type II cells in the remainder of the lung. Serum SP-D was also increased by lung inflammation. In airway inflammation induced by aerosolized ovalbumin in sensitized BALB/c mice, the serum levels were elevated after challenge. In conclusion, serum SP-D concentration is increased in mice bearing lung tumors and generally reflects the tumor burden but is also elevated during lung inflammation. PMID- 14522915 TI - Cooperative effect between immunotherapy and antiangiogenic therapy leads to effective tumor rejection in tolerant Her-2/neu mice. AB - Immunotherapy is an attractive strategy for cancer treatment. However, self tolerance is one of the major mechanisms that dampen immune responses against self-tumor antigens. We have demonstrated that Her-2/neu transgenic mice (neu mice) are tolerant to neu antigens and contain only a low avidity repertoire for neu. However, this repertoire has antitumor activity. Immunizations of neu mice are capable of activating the low-avidity T cells that, at best, retard the tumor growth. To increase the efficacy of the antitumor responses in neu mice, we hypothesized that immunotherapy in combination with antiangiogenic therapy would be a more efficient strategy for tumor eradication. The rationale for using this combination was that by decreasing the growth rate of the tumor with an antiangiogenic therapy, the low-avidity repertoire of neu mice stimulated by immunotherapeutic intervention would be more effective in destroying the slow growing tumor. To test this hypothesis, we stably expressed a soluble form of the Flt-1 vascular endothelial growth factor receptor (sFlt-1) on N202.1A cells, using a retrovirus vector. Expression of sFlt-1 on N202.1A (N202-Flt) cells significantly inhibited the tumor growth compared with N202.1A parental cells. In contrast to the application of immunotherapy alone or antiangiogenic therapy alone, which delayed the tumor growth, the combination of the two therapies provided complete inhibition of tumor growth in Her-2/neu mice. These results indicate that the use of tumor targeting with immunotherapy in simultaneous combination with antiangiogenic therapy provides a more efficient strategy for the treatment of solid tumors. PMID- 14522916 TI - Complete protection by high-dose dexamethasone against the hepatotoxicity of the novel antitumor drug yondelis (ET-743) in the rat. AB - Yondelis (ET-743) is a promising antitumor drug with hepatotoxic properties in animals and humans. Here the hypothesis was tested that dexamethasone can ameliorate manifestations of yondelis-induced hepatotoxicity in the female Wistar rat, which is the animal species with the highest sensitivity toward the adverse hepatic effect of yondelis. Hepatotoxicity was adjudged by measurement of plasma levels of alkaline phosphatase, aspartate aminotransferase, and bilirubin, and by liver histopathology. Yondelis (40 micro g/kg i.v.) alone caused a dramatic elevation of plasma alkaline phosphatase, aspartate aminotransferase, and bilirubin levels, and degeneration and patchy focal necrosis of bile duct epithelial cells. Pretreatment of rats with dexamethasone (5-20 mg/kg, p.o.) 24 h before yondelis ameliorated or abrogated the biochemical and histopathological manifestations of yondelis-induced liver changes. In contrast, when dexamethasone was administered simultaneously with yondelis, its toxicity was not reduced. Pretreatment with dexamethasone (10 mg/kg) also reversed the gene expression changes induced by yondelis in rat liver. However, dexamethasone pretreatment did not interfere with the antitumor efficacy of yondelis in rats bearing the 13762 mammary carcinoma or in four murine models. Dexamethasone (10 mg/kg) administered 24 h before yondelis decreased hepatic levels of yondelis dramatically compared with those obtained after administration of yondelis alone, whereas yondelis plasma levels after the drug combination were not markedly different from those in rats on yondelis alone. The results suggest that pretreatment with high-dose dexamethasone effectively protects rats against yondelis-mediated hepatic damage by decreasing hepatic exposure to yondelis, perhaps linked to induction of metabolism by cytochrome P450 enzymes. Pretreatment with high-dose dexamethasone should be investigated in patients who receive yondelis to ameliorate its unwanted effect on the liver. PMID- 14522917 TI - Mislocalization of membrane proteins associated with multidrug resistance in cisplatin-resistant cancer cell lines. AB - The accumulation of [(14)C]carboplatin and [(3)H]methotrexate is reduced in single-step KB epidermoid adenocarcinoma (KB-CP) cells, which are cross-resistant to carboplatin, methotrexate, and sodium arsenite. In these KB-CP cells, multidrug resistance is accompanied by mislocalization of multidrug resistance associated protein (MRP) 1 and other membrane proteins such as folate-binding protein. MRP1 was not decreased in amount in single-step variants but accumulates in a cytoplasmic fraction, and its apparent molecular weight was altered probably because of reduced glycosylation in resistant cells. This low-density compartment was partially labeled with antibodies to lectin-GSII (a Golgi marker) and Bip/GRP78 (an endoplasmic reticulum marker). Pulse-chase labeling of MRP1 with (35)S-methionine and (35)S-cysteine and pulse-chase biotinylation of cell surface MRP1 suggests that membrane protein mislocalization is caused mainly by a defect of plasma membrane protein recycling, manifested also as a defect in acidification of lysosomes. The reduced accumulation of cytotoxic compounds in the KB-CP cells is presumed to result from the failure of carrier proteins and/or transporters to localize to the plasma membrane. PMID- 14522918 TI - Consequences of telomerase inhibition and combination treatments for the proliferation of cancer cells. AB - Telomerase is expressed in most types of tumor cells but not in most somatic cells, suggesting that telomerase inhibitors may be a powerful new approach to cancer chemotherapy. Here we explore this hypothesis by treating cultured human tumor cells with a 2'-O-methoxyethyl oligonucleotide that binds the telomerase RNA template and acts as a potent inhibitor. Treatment of DU145 (Rb(-), p53(-)) and LNCaP (Rb(+), p53(+)) cells causes telomeres to shorten and cell proliferation to stop. Decreased cell proliferation in culture is not observed immediately but occurs after several weeks and is accompanied by telomere shortening. Antiproliferative effects are more profound for cells growing in soft agar or in colony formation assays, with 90% reduction in the colony-forming ability of LNCaP cells after less than 2 weeks of exposure to the inhibitor. Decreased growth of DU145 and LNCaP tumors and large reductions in prostate specific antigen levels are also observed in vivo in xenograft models. Short-term treatment of cells with telomerase inhibitors does not increase the effects of standard antiproliferative agents paclitaxel, doxorubicin, etoposide, cisplatin, or carboplatin. Long-term inhibition and telomere shortening sensitize DU145 cells, but not LNCaP cells, to cisplatin or carboplatin. These results demonstrate that methoxyethyl oligomers directed against the template region of telomerase are potent agents and that significant antiproliferative effects can be observed after 2-3 weeks of treatment. Reduced cell proliferation and tumor growth support the hypothesis that telomerase inhibition can make a useful contribution to chemotherapy and should encourage broad testing of telomerase inhibitors. PMID- 14522919 TI - Activation of peroxisome proliferator-activated receptor gamma by a novel synthetic triterpenoid 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid induces growth arrest and apoptosis in breast cancer cells. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) is a member of the nuclear hormonal receptor superfamily expressed in a large number of human cancers. Here, we demonstrate that PPARgamma is expressed and transcriptionally active in breast cancer cells independent of their p53, estrogen receptor, or human epidermal growth factor receptor 2 status. 2-Cyano-3,12-dioxooleana-1,9 dien-28-oic acid (CDDO), a novel synthetic triterpenoid, is a ligand for PPARgamma. We investigated the molecular mechanisms of CDDO on proliferation and apoptosis in breast cancer cells. In all breast cancer cell lines studied, CDDO transactivated PPARgamma, induced dose- and time-dependent cell growth inhibition, cell cycle arrest in G(1)-S and G(2)-M, and apoptosis. We then used differential cDNA array analysis to investigate the molecular changes induced by CDDO. After 16-h exposure of MCF-7 and MDA-MB-435 cells to CDDO, we found genes encoding the following proteins to be up-regulated in both cell lines: p21(Waf1/CIP1); GADD153; CAAT/enhancer binding protein transcription factor family members; and proteins involved in the ubiquitin-proteasome pathway. Among the down-regulated genes, we focused on the genes encoding cyclin D1, proliferating cell nuclear antigen, and the insulin receptor substrate 1. Using Western blot analysis and/or real-time PCR, we confirmed that CDDO regulated the expression of cyclin D1, p21(Waf1/CIP1), and Bcl-2. Cyclin D1 and p21(Waf1/CIP1) were additionally confirmed as important mediators of CDDO growth inhibition in genetically modified breast cancer cell lines. CDDO was able to significantly reduce the growth of MDA-MB-435 tumor cells in immunodeficient mice in vivo. The finding that CDDO can target genes critical for the regulation of cell cycle, apoptosis, and breast carcinogenesis suggests usage of CDDO as novel targeted therapy in breast cancer. PMID- 14522920 TI - Reactive oxygen species-dependent c-Jun NH2-terminal kinase/c-Jun signaling cascade mediates neuroblastoma cell death induced by diallyl disulfide. AB - The pharmacological properties of garlic and its derivatives are long known, and their underling mechanisms are being extensively investigated. In this study we have addressed the effects of diallyl disulfide (DADS), an oil-soluble garlic molecule, on cell growth of neuroblastoma cell SH-SY5Y, focusing on the redox events associated with this compound. Treatment of SH-SY5Y cells with DADS resulted in arrest of cell cycle in G(2)/M phase and commitment to apoptosis through the activation of the mitochondrial pathway (Bcl-2 down-regulation, cytochrome c release into the cytosol, and activation of caspase-9 and caspase 3). The earliest oxidative event observed after DADS treatment was the increase of production of reactive oxygen species, which reached the maximum yield on 30 min of DADS treatment. The oxidative burst resulted in protein and lipid damage as demonstrated by protein carbonyl accumulation and lipid peroxidation. We demonstrated that apoptosis induction was highly dependent on the activation of the redox-sensitive c-Jun NH(2)-terminal kinase (JNK)/c-Jun pathway. In particular, we established that DADS treatment induces JNK dissociation from glutathione S-transferase and its activation by phosphorylation. Moreover, treatment with JNK inhibitor I significantly reduced DADS-induced apoptosis and treatment with the spin trap 5,5'-dimethyl-1-pyrroline N-oxide or overexpression of the antioxidant enzyme copper, zinc superoxide dismutase, resulted in the inhibition of DADS-mediated toxicity through attenuation of JNK/c-Jun pathway activation. Overall, the results suggest a pivotal role for oxidative stress in DADS-induced apoptosis and, taking into account that tumor cells are deficient in antioxidants, suggest a plausible utilization of this compound as an antiproliferative agent in cancer therapy. PMID- 14522921 TI - Endothelial apoptosis initiates acute blood-brain barrier disruption after ionizing radiation. AB - Acute disruption of blood-brain barrier (BBB) is well recognized after radiation therapy to the central nervous system (CNS). We assessed the genetic regulation of acute BBB disruption and its relationship to vascular endothelial cell death in the CNS after irradiation. Adult rats were given graded single doses of X-ray to the cervical spinal cord. At different time intervals after irradiation, the irradiated spinal cord was processed for histological and immunohistochemical analysis. Disruption of blood-spinal cord barrier was assessed using albumin immunohistochemistry, i.v. injection of Evans blue dye, and (99m)Tc diethylenetriamine pentaacetic acid. In the rat spinal cord, there was a dose dependent apoptotic response during the first 24 h after irradiation, and apoptotic cells consisted of both endothelial and glial cells, as described previously (1, 2). A dose-dependent reduction in endothelial cell density was observed at 24 h after irradiation. This was associated with a similar dose dependent disruption in blood-spinal cord barrier as demonstrated by albumin immunohistochemistry. Radiation-induced apoptosis in endothelial cells has been shown to be dependent on the acid sphingomyelinase (ASMase) pathway. After a single 50-Gy dose to the cervical spinal cord of ASMase +/+ mice, there was a 47.7% reduction in endothelial cell density at 24 h compared with nonirradiated controls. No decrease in endothelial cell density was observed in irradiated ASMase -/- mice. In the irradiated spinal cord of ASMase +/+ mice, there was evidence of albumin immunoreactivity and Evans blue dye staining around microvessels, and (99m)Tc-diethylenetriamine pentaacetic acid uptake increased at 24 h. Nonirradiated controls and the irradiated spinal cord of ASMase -/- mice demonstrated no evidence of leakage. We conclude that apoptosis of endothelial cells initiates acute BBB disruption in the CNS after irradiation and that acute BBB disruption after irradiation is mediated by the ASMase pathway. PMID- 14522922 TI - Effect of interleukin 1 receptor antagonist gene transduction on human melanoma xenografts in nude mice. AB - Interleukin (IL)-1 is a pleiotropic inflammatory cytokine that promotes angiogenesis and enhances tumor growth and metastases. We evaluated the effects of IL-1 receptor antagonist (IL-1ra) on tumor growth and metastases in human melanoma xenografts. We selected two human melanoma lines (SMEL and PMEL) with differential (high versus low, respectively) constitutive production of IL-1 by ELISA. The IL-1ra gene was isolated from monocyte RNA by PCR and retrovirally transduced into SMEL and PMEL. In vitro cell proliferation was evaluated using a WST-1 assay. Athymic nude mice received s.c. or i.v. injection with parental, vector-transduced, or IL-1ra-transduced melanoma cells, and tumor growth, lung metastases, and histology were characterized. IL-1 was produced by SMEL in vitro and ex vivo (117 and 67 pg/ml/10(6) cells/24 h, respectively), but not by PMEL (15 and 0 pg/ml/10(6) cells/24 h, respectively). Neither made IL-1ra natively. Gene-transduced cell lines secreted >1000 pg/ml/10(6) cells/24 h of IL-1ra by ELISA. In vitro proliferation of each parental cell line was comparable to the proliferation rate of each transduced cell line. IL-1ra-transduced SMEL (SMEL/IL 1ra) showed significantly slower tumor growth compared with null-transduced and parental cell lines (P < 0.001, ANOVA-Bonferroni/Dunn). There was no difference in growth rates between PMEL and IL-1ra-transduced PMEL (PMEL/IL-1ra). A mixing study of SMEL and SMEL/IL-1ra showed significant inhibition of tumor growth at various ratios (P < 0.001, ANOVA-Bonferroni/Dunn). There were significantly fewer lung metastases with SMEL/IL-1ra versus SMEL (P < 0.002). IL-1ra decreases in vivo growth and metastatic potential of a human melanoma xenograft that constitutively secretes IL-1. This effect may be exploitable using clinically available IL-1ra for the treatment of human cancers. PMID- 14522923 TI - Discovery and evaluation of inhibitors of human sphingosine kinase. AB - Sphingolipid-metabolizing enzymes control the dynamic balance of the cellular levels of bioactive lipids, including the proapoptotic compound ceramide and the proliferative compound sphingosine 1-phosphate. Accumulating evidence indicates that sphingosine kinase (SK) plays a pivotal role in regulating tumor growth and that SK can act as an oncogene. Despite the importance of SK for cell proliferation, pharmacological inhibition of SK is an untested means of treating cancer because of the current lack of nonlipid inhibitors of this enzyme. To further assess the involvement of SK in human tumors, levels of RNA for SK in paired samples of cDNA prepared from tumors and normal adjacent tissue were analyzed. Expression of SK RNA was significantly elevated in a variety of solid tumors, compared with normal tissue from the same patient. To identify and evaluate inhibitors of SK, a medium throughput assay for recombinant human SK fused to glutathione S-transferase was developed, validated, and used to screen a library of synthetic compounds. A number of novel inhibitors of human SK were identified, and several representative compounds were characterized in detail. These compounds demonstrated activity at sub- to micromolar concentrations, making them more potent than any other reported SK inhibitor, and were selective toward SK compared with a panel of human lipid and protein kinases. Kinetic studies revealed that the compounds were not competitive inhibitors of the ATP binding site of SK. The SK inhibitors were antiproliferative toward a panel of tumor cell lines, including lines with the multidrug resistance phenotype because of overexpression of either P-glycoprotein or multidrug resistance phenotype 1, and were shown to inhibit endogenous human SK activity in intact cells. Furthermore, each inhibitor induced apoptosis concomitant with tumor cell cytotoxicity. Methods for the synthesis of a series of aurone inhibitors of SK were established, and a prototypical dihydroxyaurone was found to have moderate antitumor activity in vivo in the absence of overt toxicity to the mice. These compounds are the first examples of nonlipid inhibitors of SK with in vivo antitumor activity and so provide leads for additional development of inhibitors of this important molecular target. PMID- 14522924 TI - Multicellular resistance to tirapazamine is due to restricted extravascular transport: a pharmacokinetic/pharmacodynamic study in HT29 multicellular layer cultures. AB - In common with other bioreductive drugs, metabolic reduction is required for activation of the benzotriazine-di-N-oxide tirapazamine (TPZ) in hypoxic regions of tumors. This same metabolism also consumes the drug as it diffuses, impeding its penetration into hypoxic tissue. In this study, we develop a pharmacokinetic (PK)/pharmacodynamic (PD) model for TPZ that explicitly includes its diffusion characteristics as measured in multicellular layer (MCL) cultures of HT29 colon carcinoma cells. The kinetics of TPZ metabolism to its mono-N-oxide derivative SR 4317, determined by high-performance liquid chromatography using anoxic HT29 single cell suspensions, demonstrated both a first order and saturable (K(m) = 3.6 micro M) component. Cell killing, assessed by clonogenic assay under the same conditions, demonstrated an approximately quadratic concentration dependence and linear time dependence. TPZ transport through MCLs, determined under hyperoxic conditions (95% O(2)) to suppress reductive metabolism, provided a concentration independent diffusion coefficient of 0.40 x 10(-6) cm(2)s(-1). Under anoxia, this transport was strongly suppressed and was well predicted by the single cell metabolism parameters (scaled to the cell density in MCLs). These PK (transport) and PD (cytotoxicity) parameters were used to calculate cell killing as a function of distance in anoxic HT29 MCLs after the addition of TPZ to both sides of the MCL. The predicted average cell kill was in good agreement with measured values, which showed much less killing than for single cell suspensions under the same conditions. The success of this PK/PD model in predicting response in MCL shows that inefficient transport, rather than changes in intrinsic sensitivity, is responsible for TPZ resistance in these three-dimensional cell cultures and suggests that optimization of transport properties is a high priority in developing second-generation TPZ analogues. PMID- 14522925 TI - CEP-7055: a novel, orally active pan inhibitor of vascular endothelial growth factor receptor tyrosine kinases with potent antiangiogenic activity and antitumor efficacy in preclinical models. AB - Inhibition of the vascular endothelial growth factor VEGF-VEGF receptor (VEGF-R) kinase axes in the tumor angiogenic cascade is a promising therapeutic strategy in oncology. CEP-7055 is the fully synthetic orally active N,N-dimethyl glycine ester of CEP-5214, a C3-(isopropylmethoxy) fused pyrrolocarbazole with potent pan VEGF-R kinase inhibitory activity. CEP-5214 demonstrates IC(50) values of 18 nM, 12 nM, and 17 nM against human VEGF-R2/KDR kinase, VEGF-R1/FLT-1 kinase, and VEGF R3/FLT-4 kinase, respectively, in biochemical kinase assays. CEP-5214 inhibited VEGF-stimulated VEGF-R2/KDR autophosphorylation in human umbilical vein endothelial cells (HUVECs) with an IC (50) of approximately 10 nM and demonstrated an equivalent inhibition of murine FLK-1 autophosphorylation in transformed SVR endothelial cells. Evaluation of the antiangiogenic activity of CEP-5214 revealed a dose-related inhibition of microvessel growth ex vivo in rat aortic ring explant cultures and in vitro on HUVEC capillary-tube formation on Matrigel at low nanomolar concentrations. The antiangiogenic activity of CEP-5214 in these bioassays was observed in the absence of apparent cytotoxicity. Single dose p.o. or s.c. administration of CEP-7055 or CEP-5214 to CD-1 mice at 23.8 mg/kg/dose b.i.d. resulted in a reversible inhibition of VEGF-R2/FLK-1 phosphorylation in murine lung tissues. Administration p.o. of CEP-7055 at 2.57 to 23.8 mg/kg/dose b.i.d. resulted in dose-related reductions in neovascularization in vivo in porcine aortic endothelial cell (PAEC)-VEGF/basic fibroblast growth factor-Matrigel implants in nude mice (maximum, 82% inhibition), significant reductions in granuloma formation (30%) and granuloma vascularity (42%) in a murine chronic inflammation-induced angiogenesis model, and significant and sustained (6 h) inhibition of VEGF-induced plasma extravasation in rats, with an ED(50) of 20 mg/kg/dose. Chronic p.o. administration of CEP-7055 at doses of 11.9 to 23.8 mg/kg/dose b.i.d. resulted in significant inhibition (50-90% maximum inhibition relative to controls) in the growth of a variety of established murine and human s.c. tumor xenografts in nude mice, including A375 melanomas, U251MG and U87MG glioblastomas, CALU-6 lung carcinoma, ASPC-1 pancreatic carcinoma, HT-29 and HCT-116 colon carcinomas, MCF-7 breast carcinomas, and SVR angiosarcomas. Significant antitumor efficacy was observed similarly against orthotopically implanted LNCaP human prostate carcinomas in male nude mice and orthotopically implanted renal carcinoma (RENCA) tumors in BALB/c mice, in terms of a significant reduction in the metastatic score and the extent of pulmonary metastases. These antitumor responses were associated with marked increases in tumor apoptosis, and significant reductions in intratumoral microvessel density (CD34 and Factor VIII staining) of 22-38% relative to controls depending on the specific tumor xenograft. The antitumor efficacy of chronic CEP-7055 administration was independent of initial tumor volume (in the ASPC-1 pancreatic carcinoma model) and reversible on withdrawal of treatment. Chronic p.o. administration of CEP-7055 in preclinical efficacy studies for periods of up to 65 days was well tolerated with no apparent toxicity or significant morbidity. Orally administered CEP-7055 has entered Phase I clinical trials in cancer patients. PMID- 14522926 TI - Inhibition of membrane-associated calcium-independent phospholipase A2 as a potential culprit of anthracycline cardiotoxicity. AB - Administration of anthracyclines, a family of highly effective anticancer drugs, is associated with a cumulative dose-related cardiomyopathy, the etiology of which remains poorly understood. We have discovered that administration of the anthracyclines leads to a marked inhibition of membrane-associated calcium independent phospholipase A(2) (iPLA(2)) both in vitro and in vivo. To elucidate the clinical relevance of this effect and to correlate it with known cardiotoxicity of the individual anthracyclines, we have compared four anthracycline analogues: doxorubicin, daunorubicin, idarubicin, and epirubicin for their ability to inhibit iPLA(2). Isolated adult rat cardiomyocytes were treated with each analogue at concentrations of 0.1-100 micro M, and PLA(2) activity was assessed in cytosolic and membrane fractions using (16:0, [(3)H]18:1) plasmenylcholine in the absence of calcium. For all of the examined analogues, iPLA(2) inhibition was concentration and time dependent, preceded detectable changes in cell viability, and was specific to the membrane-associated enzyme. The degree of iPLA(2) inhibition by equimolar concentrations of epirubicin and idarubicin was significantly less than that of doxorubicin or daunorubicin, which correlates with the reported in vivo cardiotoxicity of these drugs. Because membrane iPLA(2) represents the majority of myocardial PLA(2) activity, its inhibition by anthracyclines would critically impair the ability of cardiomyocytes to repair oxidized phospholipids. Indeed, anthracycline-pretreated myocytes become more susceptible to the low-level oxidative stress imposed by repetitive additions of tert-butyl peroxide. The results suggest that iPLA(2) inhibition may be the initial step in a chain of events leading to chronic cardiotoxicity of the anthracyclines. PMID- 14522927 TI - Formation of tamoxifen-DNA adducts in multiple organs of adult female cynomolgus monkeys dosed with tamoxifen for 30 days. AB - The use of the antiestrogen tamoxifen (TAM) is associated with an increase in endometrial cancer. TAM-induced endometrial carcinogenesis may proceed through a genotoxin-mediated pathway, although the detection of endometrial TAM-DNA adducts in exposed women is still controversial. In this study, a monkey model has been used to investigate the question of TAM-DNA adduct formation in primates. Two methods have been used to determine TAM-DNA adducts: a TAM-DNA chemiluminescence immunoassay (TAM-DNA CIA), using an antiserum that has specificity for (E)-alpha (deoxyguanosin-N(2)-yl)-tamoxifen (dG-TAM) and (E)-alpha-(deoxyguanosin-N(2)-yl) N-desmethyltamoxifen (dG-desmethyl-TAM) and electrospray ionization tandem mass spectrometry (ES-MS/MS) coupled with on-line sample preparation and high performance liquid chromatography (HPLC). Mature (19 year old) cynomolgus monkeys were given either vehicle control (n = 1) or TAM (n = 3) twice daily for a total dose of 2 mg of TAM/kg body weight (bw)/day for 30 days by naso-gastric intubation. Tissues were harvested, and DNA was isolated from uterus, ovary, liver, brain cortex, and kidney. By TAM-DNA CIA, values for uterine TAM-DNA adducts in two monkeys were 0.9 and 1.7 adducts/10(8) nucleotides, whereas values for ovarian TAM-DNA adducts in the same animals were 0.4 and 0.5 adducts/10(8) nucleotides. Liver, brain cortex, and kidney DNA samples from the three exposed monkeys had TAM-DNA levels of 2.1-4.2 adducts/10(8) nucleotides, 0.4-5.0 adducts/10(8) nucleotides, and 0.7-2.1 adducts/10(8) nucleotides, respectively. By HPLC-ES-MS/MS, the levels of TAM-DNA adducts detected in all tissues were comparable with those observed by TAM-DNA CIA. Thus, values for uterine TAM-DNA adducts ranged from 0.5 to 1.4 adducts/10(8) nucleotides, whereas values for ovarian TAM-DNA adducts, measurable in two monkeys, were 0.2 and 0.3 adducts/10(8) nucleotides. Liver DNA contained the highest TAM-DNA adduct levels (7.0-11.1 adducts/10(8) nucleotides), whereas brain cortex DNA contained lower adduct levels (0.6-4.8 adducts/10(8) nucleotides) and the lowest levels were measured in the kidney (0.2-0.4 adducts/10(8) nucleotides). This study indicates that cynomolgus monkeys are capable of metabolizing TAM to genotoxic intermediates that form TAM-DNA adducts in multiple tissues. PMID- 14522928 TI - Identification and functional analysis of single nucleotide polymorphism in the tandem repeat sequence of thymidylate synthase gene. AB - The variable number of tandem repeat (VNTR) of thymidylate synthase (TS) gene, mainly 2 repeat (2R) and 3 repeat (3R), is one of the genetic variations that can potentially predict the effectiveness of 5-fluorouracil-based chemotherapy. In this study we identified an additional single nucleotide polymorphism (SNP) in the VNTR of TS, followed by functional and clinical analysis of the SNP. Two hundred fifty eight tumor samples were obtained from patients with primary colorectal adenocarcinoma. We observed three different patterns of electrophoresis by analysis of the VNTR with 2R/3R heterozygote. The sequencing results revealed a SNP, G/C polymorphism, within the 28-bp repeat component of TS VNTR. Each polymorphic allele was assigned as 2G, 2C, 3G, or 3C according to the combination of SNP and VNTR. Functional analysis showed that the plasmid construct with 3G sequence had three to four times greater efficiency of translation than other polymorphic sequences. 3R allele in colorectal cancer was subdivided into around half by the SNP, indicating its commonness among Japanese. TS genotypes of the patients with colorectal cancer were classified into high expression type (2R/3G, 3C/3G, and 3G/3G) and low expression type (2R/2R, 2R/3C, and 3C/3C). The patients who received oral fluoropyrimedines survived longer than the patients with no treatment in the group of low expression type. No benefit of oral fluoropyrimedines was observed in the group of high expression type. These results suggest that the double polymorphism in the TS tandem repeat sequence, the SNP and the VNTR, may provide a potential for more effective prediction of the clinical outcome of 5-fluorouracil-based chemotherapy. PMID- 14522929 TI - Radiosensitization and DNA repair inhibition by the combined use of novel inhibitors of DNA-dependent protein kinase and poly(ADP-ribose) polymerase-1. AB - The DNA repair enzymes, DNA-dependent protein kinase (DNA-PK) and poly(ADP ribose) polymerase-1 (PARP-1), are key determinants of radio- and chemo resistance. We have developed and evaluated novel specific inhibitors of DNA-PK (NU7026) and PARP-1 (AG14361) for use in anticancer therapy. PARP-1- and DNA-PK deficient cell lines were 4-fold more sensitive to ionizing radiation (IR) alone, and showed reduced potentially lethal damage recovery (PLDR) in G(0) cells, compared with their proficient counterparts. NU7026 (10 micro M) potentiated IR cytotoxicity [potentiation factor at 90% cell kill (PF(90)) = 1.51 +/- 0.04] in exponentially growing DNA-PK proficient but not deficient cells. Similarly, AG14361 (0.4 micro M) potentiated IR in PARP-1(+/+) (PF(90) = 1.37 +/- 0.03) but not PARP-1(-/-) cells. When NU7026 and AG14361 were used in combination, their potentiating effects were additive (e.g., PF(90) = 2.81 +/- 0.19 in PARP-1(+/+) cells). Both inhibitors alone reduced PLDR approximately 3-fold in the proficient cell lines. Furthermore, the inhibitor combination completely abolished PLDR. IR induced DNA double strand break (DNA DSB) repair was inhibited by both NU7026 and AG14361, and use of the inhibitor combination prevented 90% of DNA DSB rejoining, even 24-h postirradiation. Thus, there was a correlation between the ability of the inhibitors to prevent IR-induced DNA DSB repair and their ability to potentiate cytotoxicity. Thus, individually, or in combination, the DNA-PK and PARP-1 inhibitors act as potent radiosensitizers and show potential as tools for anticancer therapeutic intervention. PMID- 14522930 TI - Role of GRP58 in mitomycin C-induced DNA cross-linking. AB - Mitomycin C (MMC) is an anticancer drug that requires reductive activation to exert its toxicity. MMC is known to cross-link DNA that contributes significantly to the cytotoxicity and consequent cell death. Cytosolic NADPH:quinone oxidoreductase 1 (NQO1) and microsomal enzymes have been shown to mediate MMC induced DNA cross-linking. However, NQO1 plays only a minor role, indicating presence of other cytosolic enzymes/proteins that contribute to this process. In this study, we have characterized a unique cytosolic activity in NQO1-null mice that catalyzed MMC-induced DNA cross-linking. This activity was cofactor independent and dicoumarol insensitive. The unique cytosolic activity was purified to homogeneity. The peptide sequencing of the purified protein identified the unique cytosolic activity as GRP58 (M(r) 58,000 glucose-regulatory protein), also known as GRp57/ER60/ERp61/HIP-70/Q2 and CPT. Immunodepletion of NQO1-null mice liver cytosol and partially purified fractions with anti-GRP58 antibody led to a complete loss of GRP58 protein and consequent significant reduction of MMC-induced DNA cross-linking. Mouse cDNA encoding GRP58 was isolated and sequenced. Chinese hamster ovary cells permanently overexpressing GRP58 showed increased MMC-induced DNA cross-linking and increased cytotoxicity on exposure to MMC. Bacterially expressed and purified GRP58 increased the MMC induced DNA cross-linking when added to mouse cytosolic samples. A tissue array analysis indicated that GRP58 is ubiquitously expressed among mouse tissues, although at different levels. Expression analysis using matched human tumor/normal array revealed an up-regulation of GRP58 in breast, uterus, lung, and stomach tumors compared with normal tissues of similar origin. PMID- 14522931 TI - Suppression of microtubule dynamics by epothilone B is associated with mitotic arrest. AB - The epothilones are a group of novel microtubule-targeted, antimitotic compounds that have a paclitaxel-like, assembly enhancing effect on tubulin in vitro as well as in cultured cells. We hypothesize that epothilones induce mitotic arrest by suppressing microtubule dynamics. To test this hypothesis, we used MCF7 cells stably transfected with GFP-alpha-tubulin to analyze microtubule dynamics at three concentrations of epothilone B, one that induced no mitotic arrest (0.2 nM, 20 h), one that induced one-third maximal mitotic arrest (IC(33,) 2 nM, 20 h), and one that induced half-maximal mitotic arrest (IC(50,) 3.5 nM, 20 h). We found that epothilone B suppressed microtubule dynamics in a concentration-dependent manner coincident with mitotic block. At 0.2 nM epothilone B, dynamics were not significantly altered. At 2 nM epothilone B (IC(33)), the mean growth and shortening rates were decreased by 38 and 27%, respectively. Dynamicity was decreased by 47%. At the IC(50), 80% of the cells had nearly complete stabilization of microtubule dynamics, and no anaphase or telophase figures were observed. Comparison of the effects of epothilone B on microtubule dynamics with those of paclitaxel indicated that both drugs alter the same microtubule dynamic parameters to a similar extent. At the IC(50) for mitotic arrest, dynamicity was reduced by 54% by paclitaxel compared with 62% for epothilone B. In 65% of the cells treated with paclitaxel, the microtubules were completely stabilized. Thus, the effects of epothilone B on microtubule dynamics are remarkably similar to those of paclitaxel, suggesting that both drugs induce mitotic block by a similar mechanism. PMID- 14522932 TI - Immunological and clinical responses in women with vulval intraepithelial neoplasia vaccinated with a vaccinia virus encoding human papillomavirus 16/18 oncoproteins. AB - This study assessed the immunological and clinical responses of women with human papillomavirus (HPV) 16-associated high-grade vulval intraepithelial neoplasia (VIN) vaccinated with TA-HPV, a recombinant vaccinia virus encoding modified HPV 16 and 18 E6 and E7. Eighteen women with HPV 16-positive high-grade VIN were vaccinated with TA-HPV. The extent of their baseline disease was compared after 24 weeks by lesion measurements and histological analysis. Viral load was assessed pre- and postvaccination by real time PCR. Cell-mediated immunity to HPV 16 E6 and/or E7 peptides (HLA-A2 epitopes) or vaccinia-infected cell lysates was determined by IFN-gamma enzyme-linked immunospot (ELISPOT) and T cell proliferation using an HPV 16 L2E6E7 fusion protein. Antibodies were measured by ELISA using vaccinia-infected cell lysates or HPV 16 and 18 E6 and E7 glutathione S-transferase-fusion proteins. Lesion-infiltrating CD4(+), CD8(+), CD1a(+), and CD68(+) immune cells were assessed by immunohistochemistry. The single vaccination with TA-HPV was well tolerated, and all patients showed an increased ELISPOT and/or antibody response to vaccinia. There were significant differences in HPV-16 E7-specific ELISPOT and L2E6E7 proliferative responses in the patients at one or more time points postvaccination as compared with the prevaccination status; two patients showed transient increased antibody responses. Overall, 13 women showed an increased HPV 16-specific immune response by one or more methodologies after immunization. Eight patients demonstrated a reduction in lesion diameter of at least 50% and a further four patients showed significant symptom relief. Viral load was reduced or cleared in six of eight lesion responders but also in six of ten nonresponders. Before vaccination, clinical responders had significantly higher levels of lesion-associated CD4(+), CD8(+), and CD1a(+)-immune cells than nonresponders. There were no differences in CD68 (macrophages) between responders and nonresponders before or after vaccination. Nonresponders did show a significant increase in CD4(+)- and CD8(+)- but not CD1a(+)-immune cells postvaccination but at lower levels overall than responder patients. Local immune infiltration may be a critical factor in potential responsiveness to vaccine therapy in HPV-associated neoplasia and should be carefully monitored in future placebo-controlled trials of immunotherapy for VIN. PMID- 14522933 TI - CD4(+)CD25(+) regulatory lymphocytes require interleukin 10 to interrupt colon carcinogenesis in mice. AB - Roles for host immune response in carcinogenesis are not well defined. Recent studies have shown that microbially driven inflammation can lead to colon cancer and that prior transfer of regulatory lymphocytes expressing CD4 and CD25 prevents the innate inflammatory events that lead to colon cancer in mice. To further examine the ability of regulatory lymphocytes to inhibit carcinogenesis, 129/SvEv Rag-2-deficient mice were inoculated by gastric gavage with Helicobacter hepaticus, an enteric bacterial pathogen of mice. Mice were then treated at 1, 3, or 12 months after infection with adoptive transfer of CD4(+)CD45RB(lo)CD25(+) regulatory cells. Mice dosed with regulatory cells at 4 or 12 weeks after H. hepaticus infection had reduced severity of inflammatory bowel disease and significantly lower risk of colon cancer during the 8 month observation period, compared with infected mice that had not received cells. This suggested that regulatory cells were able to interrupt the ongoing innate immune events in the stepwise progression to cancer. Transfer of regulatory cells into chronically infected mice with established cancer reduced severity of colitis, epithelial dysplasia, and cancer, but did not eliminate all tumors. Regulatory cells lacking anti-inflammatory cytokine interleukin (IL)-10 were unable to inhibit inflammatory bowel disease, dysplasia, or cancer, showing that IL-10 was required for the protective effects of lymphocytes in this setting. Taken together, the data suggest that IL-10-mediated suppression of host innate inflammatory response was pivotal in interrupting carcinogenesis. Regulatory lymphocytes and cytokines may have implications for novel therapies for colon cancer in humans. PMID- 14522934 TI - On the role of unmutated antigens in tumor rejection in mice with unperturbed T cell repertoires. AB - We investigate here whether P1A, which was the first CTL-recognized and unmutated tumor antigen to be identified, is a tumor rejection antigen for J558 plasmacytoma in mice with an unperturbed T-cell repertoire. We show that although transgenic mice expressing P1A in the thymus have almost complete deletion of P1A reactive T cells, they reject the B7-1-transfected J558 at a rate comparable with wild-type mice. Thus, P1A is not a necessary tumor rejection antigen for the J558 tumor cells. On the other hand, if anti-P1A CTL response is sufficient for tumor rejection, tumor cells must lose the antigenic epitope to evade CTL destruction. To test this, we analyze whether tumor cells escaping J558-B7 immune spleen cells harbor mutations in the P1A epitope. We find that although the spleen contained a high proportion of P1A-reactive T cells, the recurrent tumor cells have no mutation in the P1A antigenic epitope and remain susceptible to lysis by P1CTL. Thus, the antigen-bearing tumor cells can evade immune destruction in the presence of a high number of P1A-reactive T cells. Taken together, our results demonstrate that in mice with a normal TCR repertoire, substantial numbers of P1A reactive T cells are neither necessary nor sufficient for tumor rejection and raise interesting questions regarding the significance of T-cell response against unmutated tumor antigens. PMID- 14522936 TI - Stroma adjacent to metastatic mature teratoma after chemotherapy for testicular germ cell tumors is derived from the same progenitor cells as the teratoma. AB - Metastatic mature teratoma is often present in postchemotherapy surgical specimens of lymph nodes from patients with pathological stage II or III testicular germ cell tumors. The stromal cells in these lesions have generally been considered "fibrosis" secondary to the chemotherapy and the necrosis it causes, although the frequent cytological atypia of the stromal cells suggests that they may be neoplastic. We studied 25 patients with pathological stage II or III testicular cancer who were treated with platinum-based chemotherapy followed by surgical resection of retroperitoneal lymph nodes that contained metastatic mature teratoma with "fibrosis" to determine the reactive or neoplastic nature of the stromal cells. We compared the pattern of allelic loss using nine microsatellite DNA markers (D9S177, D9S303, D9S778, D9S171, D12S1015, D1S508, D2S156, D18S46, and D11S903) between the epithelial cells of the teratoma and the cells in the adjacent stroma. A laser capture microdissection technique facilitated preparation of genomic DNA from the epithelial components of teratoma, adjacent stromal cells, and normal lymph node tissue from each patient. Of the 25 patients, loss of heterozygosity was seen at a minimum of one focus in 22 (92%) of the teratoma specimens and 16 (64%) of the adjacent stroma. Of the 16 cases for which the stroma showed loss of heterozygosity, 8 cases showed the identical pattern of allelic loss in the epithelial cells of the adjacent teratoma at all nine DNA loci studied. The remaining eight cases showed similar allelic loss in at least one of the nine DNA loci analyzed. Interestingly, three cases showed loss of heterozygosity in the stroma that was not seen in the matching teratoma specimens. Our results indicate that the stromal cells adjacent to metastatic mature teratoma in postchemotherapy lymph node specimens frequently have genetic abnormalities similar to the metastatic teratoma. Concordant genetic alterations observed in teratoma and stroma suggest that both are derived from the same element of the original germ cell tumor or the same progenitor cell. PMID- 14522935 TI - The selective estrogen receptor modulator trioxifene (LY133314) inhibits metastasis and extends survival in the PAIII rat prostatic carcinoma model. AB - Trioxifene (LY133314) is a selective estrogen receptor modulator (SERM) with competitive binding activity against estradiol for estrogen receptor alpha (ERalpha) and antagonistic activity against ERalpha-mediated gene expression. The PAIII rat prostatic adenocarcinoma (PCa) is an androgen receptor-negative, ERalpha- and ERbeta-positive, spontaneously metastatic rodent tumor cell line. After s.c. implantation of 10(6) PAIII cells in the tail, s.c. administration of trioxifene (2.0, 4.0, 20.0, or 40.0 mg/kg-day) for 30 days produced significant (P < 0.05) inhibition of PAIII metastasis from the primary tumor in the tail to the gluteal and iliac lymph nodes (maximum nodal weight decreases, 86% and 88% from control values, respectively). PAIII metastasis to the lungs was significantly inhibited by trioxifene treatment. Numbers of pulmonary foci in PAIII-bearing rats were significantly (P < 0.05) reduced by trioxifene administration in a dose-related manner (maximal reduction, 98% from control values). Continual administration of the compound significantly (P < 0.05) extended survival of PAIII-bearing rats. Trioxifene inhibited the proliferation of PAIII cells at micromolar levels in vitro but did not slow growth of the primary tumor growth in the tail. Trioxifene administration also produced regression of male accessory sex organs. In PAIII-tumor-bearing animals, trioxifene administration produced a maximal regression of 76% for ventral prostate and 64% for seminal vesicle (P < 0.05 for both). SERMs may be preferable to estrogens given their efficacy in experimental PCa models and relative lack of side effects observed in clinical trials. Our data support the contention that trioxifene represents a SERM with potential antimetastatic efficacy for the treatment of androgen-independent, metastatic PCa. PMID- 14522937 TI - Disease-associated expression profiles in peripheral blood mononuclear cells from patients with advanced renal cell carcinoma. AB - Expression profiling has demonstrated that transcriptomes of primary malignancies differ from those in normal tissue. It is unknown, however, whether there exist "surrogate" transcriptional markers in peripheral blood mononuclear cells (PBMCs) of patients with solid tumors. We identified transcripts expressed differentially between PBMCs from renal cell carcinoma patients and normal subjects, some of which appear to reflect specific immune responses of circulating cells. We also identified small sets of predictor genes distinguishing PBMCs from renal cell carcinoma patients and normal volunteers with high accuracy. The present findings have important implications for diagnosis and future clinical pharmacogenomic studies of antitumor therapies. PMID- 14522938 TI - NY-ESO-1 and LAGE-1 cancer-testis antigens are potential targets for immunotherapy in epithelial ovarian cancer. AB - Cancer-testis (CT) antigens are expressed in a variety of cancers, but not in normal adult tissues, except for germ cells of the testis, and hence appear to be ideal targets for immunotherapy. In an effort to examine the potential of NY-ESO 1 and LAGE-1 CT antigens for immunotherapy in epithelial ovarian cancer (EOC), we examined the expression of these antigens by reverse transcription-PCR (RT-PCR) and immunohistochemistry (IHC) in a large panel of EOC tissues and cell lines. Sera from a subgroup of the patients were tested for NY-ESO-1/LAGE-1 antibody by ELISA. The data indicated that four ovarian cancer cell lines were positive for one or both CT antigens. Expression of NY-ESO-1 in EOC was demonstrated by RT-PCR and/or IHC in 82 of 190 (43%) specimens. NY-ESO-1 expression by IHC ranged from homogeneous to heterogeneous pattern. LAGE-1 mRNA expression was present in 22 of 107 (21%) tumor tissues. Overall, the expression of either NY-ESO-1 or LAGE-1 mRNA was present in 42 of 107 (40%) EOC specimens and coexpression of both antigens was demonstrated in 11% of specimens. Antibody to NY-ESO-1/LAGE-1 was present in 11 of 37 (30%) patients whose tumors expressed either NY-ESO-1 or LAGE 1. Detectable antibodies were present for up to 3 years after initial diagnosis. Although there was no statistically significant relation between expression of NY ESO-1/LAGE-1 antigen and survival, the data showed aberrant expression of NY-ESO 1 and LAGE-1 by IHC/RT-PCR in a significant proportion of EOC patients. These findings indicate that NY-ESO-1 and LAGE-1 are attractive targets for antigen specific immunotherapy in EOC. PMID- 14522939 TI - Overexpression of endocan induces tumor formation. AB - Endocan is a proteoglycan specifically secreted by endothelial cells. Through its glycan domains, endocan binds to hepatocyte growth factor and increases its mitogenic activity. Here, we show that human embryonic kidney 293 cells, which have been genetically engineered to overexpress endocan, form tumors when injected s.c. in SCID mice. Both the glycan and a phenylalanine-rich region of endocan are necessary for mediating tumor growth activity. Blocking the phenylalanine-rich region with a monoclonal antibody results in a marked reduction of tumor growth. Finally, we report that circulating levels of endocan are increased in mice with the endocan-expressing human embryonic kidney 293 cell tumors and in a series of adult patients with lung cancer. Taken together, these results suggest that (a) endothelial-derived endocan induces tumor growth, (b) antibodies to endocan may have therapeutic potential, and (c) circulating levels of endocan may eventually represent a novel marker for cancer. PMID- 14522940 TI - Concomitant suppression of hyperlipidemia and intestinal polyp formation in Apc deficient mice by peroxisome proliferator-activated receptor ligands. AB - Epidemiological studies have shown a positive association of colon cancer with hyperlipidemia. Furthermore, signaling generated by peroxisome proliferator activated receptor (PPAR) alpha and gamma ligands, suggested to be candidate tumor preventive agents, has been shown to lower serum triglyceride levels. In the present study, we assessed hyperlipidemia in Apc-deficient mice, model animals for human familial adenomatous polyposis, and examined the effects of pioglitazone and bezafibrate, respectively, PPARgamma and PPARalpha agonists, on both hyperlipidemia and intestinal polyposis. Serum lipid levels in Apc(1309) mice and Min mice from 6 to 15 weeks of age were measured. Although serum levels of triglyceride and cholesterol were low in both Apc(1309) and wild-type mice at 6 weeks, triglycerides were elevated 10-fold in Apc(1309) mice by the age of 12 weeks but not in their wild-type counterparts. Cholesterol was also increased significantly, and marked centrilobular-restricted steatosis was observed in the livers of aged Apc(1309) mice. Similar findings were observed for Min mice at 15 weeks of age. Moreover, lipoprotein lipase mRNA levels in the liver and small intestine of Apc(1309) and Min mice were demonstrated to be lower than those in wild-type mice. Treatment of Apc(1309) mice with 100 and 200 ppm pioglitazone or bezafibrate for 6 weeks from 6 weeks of age caused dose-dependent reduction in serum triglycerides and cholesterol, along with reduction in the numbers of intestinal polyps to 67% of the control value. The present study clearly demonstrated a hyperlipidemic state in Apc gene-deficient mice and a potential of PPARalpha and PPARgamma ligands to suppress both hyperlipidemia and polyp formation. Hyperlipidemia in these mice may thus be associated with their intestinal lesion development. PMID- 14522941 TI - Breast cancer and nonsteroidal anti-inflammatory drugs: prospective results from the Women's Health Initiative. AB - We analyzed data from the prospective Women's Health Initiative (WHI) Observational Study to examine the effects of regular use of aspirin, ibuprofen, and other nonsteroidal anti-inflammatory drugs (NSAIDs) on breast cancer risk. We studied a population of 80,741 postmenopausal women between 50 and 79 years of age who reported no history of breast cancer or other cancers (excluding nonmelanoma skin cancer), and we completed a personal baseline interview that elicited comprehensive health information including data on breast cancer risk factors and NSAID use. All of the cases were adjudicated by WHI physicians using pathology reports. Our analysis was based on 1392 confirmed cases of breast cancer. Relative risks (RRs) with 95% confidence intervals (CIs) were estimated with adjustment for age and other breast cancer risk factors. Regular NSAID use (two or more tablets/week) for 5-9 years produced a 21% reduction in the incidence of breast cancer (RR, 0.79; 95% CI, 0.60-1.04); regular NSAID use for 10 or more years produced a 28% reduction (RR, 0.72; CI, 0.56-0.91), and there was a statistically significant inverse linear trend of breast cancer incidence with the duration of NSAID use (P < 0.01). The estimated risk reduction for long term use of ibuprofen (RR, 0.51; CI, 0.28-0.96) was greater than for aspirin (RR, 0.79; CI, 0.60-1.03). Subgroup analysis by breast cancer risk factors did not result in effect modification. Regular use of acetaminophen (an analgesic agent with little or no anti-inflammatory activity) or low-dose aspirin (<100 mg) was unrelated to the incidence of breast cancer. Our results indicate that the regular use of aspirin, ibuprofen, or other NSAIDs may have a significant chemopreventive effect against the development of breast cancer and underscore the need for clinical trials to confirm this effect. PMID- 14522942 TI - Which end: dissecting Ku's function at telomeres and double-strand breaks. PMID- 14522943 TI - Degradation of targeted mRNAs in Escherichia coli: regulation by a small antisense RNA. PMID- 14522944 TI - NF-kappaB mediates inhibition of mesenchymal cell differentiation through a posttranscriptional gene silencing mechanism. AB - Cytokines, such as tumor necrosis factor-alpha (TNFalpha), potently inhibit the differentiation of mesenchymal cells and down-regulate the expression of Sox9 and MyoD, transcription factors required for chondrocyte and myocyte development. Previously, we demonstrated that NF-kappaB controls TNFalpha-mediated suppression of myogenesis through a mechanism involving MyoD mRNA down-regulation. Here, we show that NF-kappaB also suppresses chondrogenesis and destabilizes Sox9 mRNA levels. Multiple copies of an mRNA cis-regulatory motif (5'-ACUACAG-3') are necessary and sufficient for NF-kappaB-mediated Sox9 and MyoD down-regulation. Thus, in response to cytokine signaling, NF-kappaB modulates the differentiation of mesenchymal-derived cell lineages via RNA sequence-dependent, posttranscriptional down-regulation of key developmental regulators. PMID- 14522945 TI - mSin3-associated protein, mSds3, is essential for pericentric heterochromatin formation and chromosome segregation in mammalian cells. AB - The histone code guides many aspects of chromosome biology including the equal distribution of chromosomes during cell division. In the chromatin domains surrounding the centromere, known as pericentric heterochromatin, histone modifications, particularly deacetylation and methylation, appear to be essential for proper chromosome segregation. However, the specific factors and their precise roles in this highly orchestrated process remain under active investigation. Here, we report that germ-line or somatic deletion of mSds3, an essential component of the functional mSin3/HDAC corepressor complex, generates a cell-lethal condition associated with rampant aneuploidy, defective karyokinesis, and consequently, a failure of cytokinesis. mSds3-deficient cells fail to deacetylate and methylate pericentric heterochromatin histones and to recruit essential heterochromatin-associated proteins, resulting in aberrant associations among heterologous chromosomes via centromeric regions and consequent failure to properly segregate chromosomes. Mutant mSds3 molecules that are defective in mSin3 binding fail to rescue the mSds3 null phenotypes. On the basis of these findings, we propose that mSds3 and its associated mSin3/HDAC components play a central role in initiating the cascade of pericentric heterochromatin-specific modifications necessary for the proper distribution of chromosomes during cell division in mammalian cells. PMID- 14522946 TI - Inheritance of Polycomb-dependent chromosomal interactions in Drosophila. AB - Maintenance of cell identity is a complex task that involves multiple layers of regulation, acting at all levels of chromatin packaging, from nucleosomes to folding of chromosomal domains in the cell nucleus. Polycomb-group (PcG) and trithorax-group (trxG) proteins maintain memory of chromatin states through binding at cis-regulatory elements named PcG response elements or cellular memory modules. Fab-7 is a well-defined cellular memory module involved in regulation of the homeotic gene Abdominal-B (Abd-B). In addition to its action in cis, we show here by three-dimensional FISH that the Fab-7 element leads to association of transgenes with each other or with the endogenous Fab-7, even when inserted in different chromosomes. These long-distance interactions enhance PcG-mediated silencing. They depend on PcG proteins, on DNA sequence homology, and on developmental progression. Once long-distance pairing is abolished by removal of the endogenous Fab-7, the derepressed chromatin state induced at the transgene locus can be transmitted through meiosis into a large fraction of the progeny, even after reintroduction of the endogenous Fab-7. Strikingly, meiotic inheritance of the derepressed state involves loss of pairing between endogenous and transgenic Fab-7. This suggests that transmission of nuclear architecture through cell division might contribute to inheritance of chromatin states in eukaryotes. PMID- 14522947 TI - KNL-1 directs assembly of the microtubule-binding interface of the kinetochore in C. elegans. AB - Segregation of the replicated genome during cell division requires kinetochores, mechanochemical organelles that assemble on mitotic chromosomes to connect them to spindle microtubules. CENP-A, a histone H3 variant, and CENP-C, a conserved structural protein, form the DNA-proximal foundation for kinetochore assembly. Using RNA interference-based genomics in Caenorhabditis elegans, we identified KNL-1, a novel kinetochore protein whose depletion, like that of CeCENP-A or CeCENP-C, leads to a "kinetochore-null" phenotype. KNL-1 is downstream of CeCENP A and CeCENP-C in a linear assembly hierarchy. In embryonic extracts, KNL-1 exhibits substoichiometric interactions with CeCENP-C and forms a near stoichiometric complex with CeNDC-80 and HIM-10, the C. elegans homologs of Ndc80p/HEC1p and Nuf2p-two widely conserved outer kinetochore components. However, CeNDC-80 and HIM-10 are not functionally equivalent to KNL-1 because their inhibition, although preventing formation of a mechanically stable kinetochore-microtubule interface and causing chromosome missegregation, does not result in a kinetochore-null phenotype. The greater functional importance of KNL 1 may be due to its requirement for targeting multiple components of the outer kinetochore, including CeNDC-80 and HIM-10. Thus, KNL-1 plays a central role in translating the initiation of kinetochore assembly by CeCENP-A and CeCENP-C into the formation of a functional microtubule-binding interface. PMID- 14522948 TI - Local regulation of fat metabolism in peripheral nerves. AB - We comprehensively analyzed gene expression during peripheral nerve development by performing microarray analyses of premyelinating, myelinating, and postmyelinating mouse sciatic nerves, and we generated a database of candidate genes to be tested in mapped peripheral neuropathies. Unexpectedly, we identified a large cluster of genes that are (1) maximally expressed only in the mature nerve, after myelination is complete, and (2) tied to the metabolism of storage (energy) lipids. Many of these late-onset genes are expressed by adipocytes, which we find constitute the bulk of the epineurial compartment of the adult nerve. However, several such genes, including SREBP-1, SREBP-2, and Lpin1, are also expressed in the endoneurium. We find that Lpin1 null mutations lead to lipoatrophy of the epineurium, and to the dysregulation of a battery of genes required for the regulation of storage lipid metabolism in both the endoneurium and peri/epineurium. Together with the observation that these mutations also result in peripheral neuropathy, our findings demonstrate a crucial role for local storage lipid metabolism in mature peripheral nerve function, and have important implications for the understanding and treatment of peripheral neuropathies that are commonly associated with metabolic diseases such as lipodystrophy and diabetes. PMID- 14522949 TI - Beta1 integrins regulate chondrocyte rotation, G1 progression, and cytokinesis. AB - Beta1 integrins are highly expressed on chondrocytes, where they mediate adhesion to cartilage matrix proteins. To assess the functions of beta1 integrin during skeletogenesis, we inactivated the beta1 integrin gene in chondrocytes. We show here that these mutant mice develop a chondrodysplasia of various severity. beta1 deficient chondrocytes had an abnormal shape and failed to arrange into columns in the growth plate. This is caused by a lack of motility, which is in turn caused by a loss of adhesion to collagen type II, reduced binding to and impaired spreading on fibronectin, and an abnormal F-actin organization. In addition, mutant chondrocytes show decreased proliferation caused by a defect in G1/S transition and cytokinesis. The G1/S defect is, at least partially, caused by overexpression of Fgfr3, nuclear translocation of Stat1/Stat5a, and up-regulation of the cell cycle inhibitors p16 and p21. Altogether these findings establish that beta1-integrin-dependent motility and proliferation of chondrocytes are mandatory events for endochondral bone formation to occur. PMID- 14522950 TI - ELAV inhibits 3'-end processing to promote neural splicing of ewg pre-mRNA. AB - The embryonic lethal abnormal visual system (ELAV) is a gene-specific regulator of alternative pre-mRNA processing in neurons of Drosophila. Here we define a functional in vivo binding site for ELAV in neurons through the development of a reporter gene system in transgenic animals in combination with in vitro binding assays. ELAV binds to erect wing (ewg) RNA 3' of a polyadenylation site in the terminal intron 6. At this polyadenylation site, ELAV inhibits 3'-end processing in vitro in a dose-dependent and sequence-specific manner, and ELAV binding is necessary in vivo to promote splicing of ewg intron 6. Further, the AAUAAA poly(A) complex recognition sequence, together with ELAV, is required to regulate neural 3' splice site choice in vivo. In addition, the use of segmentally labeled RNA substrates in UV cross-linking assays suggest that ELAV does not inhibit or redirect binding of cleavage factor dCstF64 at the regulated polyadenylation site on ewg RNA. These data indicate that binding of 3'-end processing factors, together with ELAV, can regulate alternative splicing. PMID- 14522951 TI - Recognition and repair of the cyclobutane thymine dimer, a major cause of skin cancers, by the human excision nuclease. AB - The cyclobutane thymine dimer is the major DNA lesion induced in human skin by sunlight and is a primary cause of skin cancer, the most prevalent form of cancer in the Northern Hemisphere. In humans, the only known cellular repair mechanism for eliminating the dimer from DNA is nucleotide excision repair. Yet the mechanism by which the dimer is recognized and removed by this repair system is not known. Here we demonstrate that the six-factor human excision nuclease recognizes and removes the dimer at a rate consistent with the in vivo rate of removal of this lesion, even though none of the six factors alone is capable of efficiently discriminating the dimer from undamaged DNA. We propose a recognition mechanism by which the low-specificity recognition factors, RPA, XPA, and XPC, act in a cooperative manner to locate the lesion and, aided by the kinetic proofreading provided by TFIIH, form a high-specificity complex at the damage site that initiates removal of thymine dimers at a physiologically relevant rate and specificity. PMID- 14522952 TI - STAT3-dependent enhanceosome assembly and disassembly: synergy with GR for full transcriptional increase of the alpha 2-macroglobulin gene. AB - We describe a detailed time course of the assembly and disassembly of a STAT3 dependent, glucocorticoid-supplemented enhanceosome for the alpha2-macroglobulin (alpha2-M) gene and compare this with a detailed time course of transcription of the gene by run-on analysis. The glucocorticoid receptor (GR) can associate with the enhanceosome without STAT3. Furthermore, the enhanceosome contains c-Jun/c Fos and OCT-1 constitutively. All of these factors (GR, c-Jun, OCT-1) have transcription activation domains, but STAT3 is required for the observed transcriptional increase. The time course of enhanceosome occupation by GR and tyrosine-phosphorylated STAT3 shows that these transcription factors precede by approximately 5-10 min the arrival of RNA polymerase II (Pol II). The enhanceosome remains assembled for approximately 90 min in the continued presence of both inducers. When IL-6 and Dex are removed (after 30 min of treatment), the disappearance within an additional 30 min of the established enhanceosome indicates that renewal of STAT3 and GR binding must occur in the continued presence of IL-6+Dex. Compared with the total nuclear tyrosine-phosphorylated STAT3 capable of binding DNA, the chromatin-associated STAT3 resists dephosphorylation and appears to recycle to maintain the enhanceosome. Run-on transcription shows a lag after full enhanceosome occupation that can be largely but not completely explained by the approximately 30 min transit time of Pol II across the alpha2-Mlocus. PMID- 14522953 TI - Alternative trans-splicing of constant and variable exons of a Drosophila axon guidance gene, lola. AB - longitudinals lacking (lola) is a complex Drosophila gene encoding at least 20 protein isoforms,each bearing the same N-terminal constant region linked to a different C-terminal variable region. Different isoforms specify different aspects of axon growth and guidance. We show here that lola mRNAs are generated by alternative trans-splicing of exons sequentially encoded by the same DNA strand. Chromosomal pairing facilitates interallelic trans-splicing,allowing complementation between mutations in the constant and those in the variable exons. We demonstrate that at least one variable exon is transcribed from its own promoter,and trans-spliced to the constant exons transcribed separately. PMID- 14522954 TI - Human pyridoxal phosphatase. Molecular cloning, functional expression, and tissue distribution. AB - Pyridoxal phosphatase catalyzes the dephosphorylation of pyridoxal 5'-phosphate (PLP) and pyridoxine 5'-phosphate. A human brain cDNA clone was identified to the PLP phosphatase on the basis of peptide sequences obtained previously. The cDNA predicts a 296-amino acid protein with a calculated Mr of 31698. The open reading frame is encoded by two exons located on human chromosome 22q12.3, and the exon intron junction contains the GT/AG consensus splice site. In addition, a full length mouse PLP phosphatase cDNA of 1978 bp was also isolated. Mouse enzyme encodes a protein of 292 amino acids with Mr of 31512, and it is localized on chromosome 15.E1. Human and mouse PLP phosphatase share 93% identity in protein sequence. A BLAST search revealed the existence of putative proteins in organism ranging from bacteria to mammals. Catalytically active human PLP phosphatase was expressed in Escherichia coli, and characteristics of the recombinant enzyme were similar to those of erythrocyte enzyme. The recombinant enzyme displayed Km and kcat values for pyridoxal of 2.5 microM and 1.52 s(-1), respectively. Human PLP phosphatase mRNA is differentially expressed in a tissue-specific manner. A single mRNA transcript of 2.1 kb was detected in all human tissues examined and was highly abundant in the brain. Obtaining the molecular properties for the human PLP phosphatase may provide new direction for investigating metabolic pathway involving vitamin B6. PMID- 14522955 TI - Identification of an N-domain histidine essential for chaperone function in calreticulin. AB - Calreticulin is an endoplasmic reticulum (ER) luminal Ca(2+)-binding chaperone involved in folding of newly synthesized glycoproteins via the "calreticulin calnexin cycle." We reconstituted ER of calreticulin-deficient cells with N terminal histidine (His25, His82, His128, and His153) calreticulin mutants and carried out a functional analysis. In crt(-/-) cells bradykinin-dependent Ca2+ release is altered, and the reestablishment of bradykinin-dependent Ca2+ release was used as a marker for calreticulin function. Bradykinin-dependent Ca2+ release from the ER was rescued by wild type calreticulin and by the His25, His82, or His128 mutant but not by the His153 mutant. Wild type calreticulin and the His25, His82, and His128 mutants all prevented in vitro thermal aggregation of malate dehydrogenase and IgY, whereas the His153 mutant did not, indicating that His153 chaperone function was impaired. Biophysical analysis of His153 mutant revealed that conformation changes in calreticulin mutant may be responsible for the loss of its chaperone activity. We conclude that mutation of a single amino acid residue in calreticulin has devastating consequences for its chaperone function, indicating that mutations in chaperones may play a significant role in protein folding disorders. PMID- 14522956 TI - Role of disulfide bonds in Acrp30/adiponectin structure and signaling specificity. Different oligomers activate different signal transduction pathways. AB - Acrp30/adiponectin is an adipocyte-derived serum protein with important roles in regulation of lipid and glucose metabolism, but which of its isoforms are biologically active remains controversial. We addressed this issue by first characterizing the structure of each individual Acrp30 oligomer and the determinants responsible for multimer formation. Freeze etch electron microscopy showed the trimer to exhibit a ball-and- stick-like structure containing a large globular sphere, an extended collagen stalk, and a smaller sphere on the opposite end of the stalk. The hexamer consists of two adjacent trimeric globular domains and a single stalk composed of collagen domains from two trimers. Although not necessary for trimer formation or stability, two of the three monomers in an Acrp30 trimer are covalently linked by a disulfide bond between cysteine residues at position 22. In contrast, assembly of hexameric and higher molecular weight (HMW) forms of Acrp30 depends upon formation of Cys22-mediated disulfide bonds because their reduction with dithiothreitol or substitution of Cys22 with alanine led exclusively to trimers. HMW and hexamer isoforms of Acrp30 activated NF kappaB in C2C12 cells, but trimers, either natural, formed by reduction of Acrp30 hexamer, or formed by the C22A mutant, did not. In contrast, incubation of isolated rat extensor digitorum longus with naturally formed Acrp30 trimers or trimeric C22A Acrp30 led to increased phosphorylation of AMP-activated protein kinase-alpha at Thr172 and its activation. Hexameric and HMW Acrp30 could not activate AMP-activated protein kinase. Thus, trimeric and HMW/hexameric Acrp30 activate different signal transduction pathways, and Acrp30 represents a novel example of the control of ligand signaling via changes in its oligomerization state. PMID- 14522957 TI - How nerve growth factor drives physiological and inflammatory expressions of acid sensing ion channel 3 in sensory neurons. AB - Nerve growth factor (NGF) is a key element of inflammatory pain. It induces hyperalgesia by up-regulating the transcription of genes encoding receptors, ion channels, and neuropeptides. Acid-sensing ion channel 3 (ASIC3), a depolarizing sodium channel gated by protons during tissue acidosis, is specifically expressed in sensory neurons. It has been associated to cardiac ischemic and inflammatory pains. We previously showed that low endogenous NGF was responsible for ASIC3 basal expression and high NGF during inflammation increased ASIC3 expression parallely to the development of neuron hyperexcitability associated with hyperalgesia. NGF is known to activate numerous signaling pathways through trkA and p75 receptors. We now show that (i). NGF controls ASIC3 basal expression through constitutive activation of a trkA/phospholipase C/protein kinase C pathway, (ii). high inflammatory-like NGF induces ASIC3 overexpression through a trkA/JNK/p38MAPK pathway and a p75-dependent mechanism as a transcriptional switch, and (iii). NGF acts through AP1 response elements in ASIC3 encoding gene promoter. These new data indicate potential targets that could be used to develop new treatments against inflammatory pain. PMID- 14522958 TI - Post-transcriptional control of human maxiK potassium channel activity and acute oxygen sensitivity by chronic hypoxia. AB - Various cardiorespiratory diseases (e.g. congestive heart failure, emphysema) result in systemic hypoxia and patients consequently demonstrate adaptive cellular responses which predispose them to conditions such as pulmonary hypertension and stroke. Central to many affected excitable tissues is activity of large conductance, Ca2+-activated K+ (maxiK) channels. We have studied maxiK channel activity in HEK293 cells stably co-expressing the most widely distributed of the human alpha- and beta-subunits that constitute these channel following maneuvers which mimic severe hypoxia. At all [Ca2+]i, chronic hypoxia (approximately 18 mm Hg, 72 h) increased K+ current density, most markedly at physiological [Ca2+]i K+ currents in cells cultured in normoxia showed a [Ca2+]i dependent sensitivity to acute hypoxic inhibition ( approximately 25 mm Hg, 3 min). However, chronic hypoxia dramatically changed the Ca2+ sensitivity of this acute hypoxic inhibitory profile such that low [Ca2+]i could sustain an acute hypoxic inhibitory response. Chronic hypoxia caused no change in alpha-subunit immunoreactivity with Western blotting but evoked a 3-fold increase in beta subunit expression. These observations were fully supported by immunocytochemistry, which also suggested that chronic hypoxia augmented alpha/beta-subunit co-localization at the plasma membrane. Using a novel nuclear run-on assay and RNase protection we found that chronic hypoxia did not alter mRNA production rates or steady-state levels, which suggests that this important environmental cue modulates maxiK channel function via post-transcriptional mechanisms. PMID- 14522959 TI - Enhanced Akt signaling is an early pro-survival response that reflects N-methyl-D aspartate receptor activation in Huntington's disease knock-in striatal cells. AB - Huntington's disease features the loss of striatal neurons that stems from a disease process that is initiated by mutant huntingtin. Early events in the disease cascade, which predate overt pathology in Hdh CAG knock-in mouse striatum, implicate enhanced N-methyl-D-aspartate (NMDA) receptor activation, with excitotoxity caused by aberrant Ca2+ influx. Here we demonstrate in precise genetic Huntington's disease mouse and striatal cell models that these early phenotypes are associated with activation of the Akt pro-survival signaling pathway. Elevated levels of activated Ser(P)473-Akt are detected in extracts of Hdh(Q111/Q111) striatum and cultured mutant STHdh(Q111/Q111) striatal cells, compared with their wild type counterparts. Akt activation in mutant striatal cells is associated with increased Akt signaling via phosphorylation of GSK3beta at Ser9. Consequent decreased turnover of transcription co-factor beta-catenin leads to increased levels of beta-catenin target gene cyclin D1. Akt activation is phosphatidylinositol 3-kinase dependent, as demonstrated by increased levels of Ser(P)241-PDK1 kinase and decreased Ser(P)380-PTEN phosphatase. Moreover, Akt activation can be normally stimulated by treatment with insulin growth factor-1 and blocked by treatment with the phosphatidylinositol 3-kinase inhibitor LY294002. However, in contrast to wild type cells, Akt activation in mutant striatal cells can be blocked by the addition of the NMDA receptor antagonist MK 801. Akt activation in mutant striatal cells is Ca(2+)-dependent, because treatment with EGTA reduces levels of Ser(P)473-Akt. Thus, consistent with excitotoxicity early in the disease process, activation of the Akt pro-survival pathway in mutant knock-in striatal cells predates overt pathology and reflects mitochondrial dysfunction and enhanced NMDA receptor signaling. PMID- 14522960 TI - The low density lipoprotein receptor-related protein modulates protease activity in the brain by mediating the cellular internalization of both neuroserpin and neuroserpin-tissue-type plasminogen activator complexes. AB - Proteases contribute to a variety of processes in the brain; consequently, their activity is carefully regulated by protease inhibitors, such as neuroserpin. This inhibitor is thought to be secreted by axons at synaptic regions where it controls tissue-type plasminogen activator (tPA) activity. Mechanisms regulating neuroserpin are not known, and the current studies were undertaken to define the cellular pathways involved in neuroserpin catabolism. We found that both active neuroserpin and neuroserpin.tPA complexes were internalized by mouse cortical cultures and embryonic fibroblasts in a process mediated by the low density lipoprotein receptor-related protein (LRP). Surprisingly, despite the fact that active neuroserpin is internalized by LRP, this form of the molecule does not directly bind to LRP on its own, indicating the requirement of a cofactor for neuroserpin internalization. Our studies ruled out the possibility that endogenously produced plasminogen activators (i.e. tPA and urokinase-type plasminogen activator) are responsible for the LRP-mediated internalization of active neuroserpin, but could not rule out the possibility that another cell associated proteases capable of binding active neuroserpin functions in this capacity. In summary, neuroserpin levels appear to be carefully regulated by LRP and an unidentified cofactor, and this pathway may be critical for maintaining the balance between proteases and inhibitors. PMID- 14522961 TI - CEACAM1, a cell-cell adhesion molecule, directly associates with annexin II in a three-dimensional model of mammary morphogenesis. AB - The epithelial cell adhesion molecule CEACAM1 (carcinoembryonic antigen cell adhesion molecule-1) is down-regulated in colon, prostate, breast, and liver cancer. Here we show that CEACAM1-4S, a splice form with four Ig-like ectodomains and a short cytoplasmic domain (14 amino acids), directly associates with annexin II, a lipid raft-associated molecule, which is also down-regulated in many cancers. Annexin II was identified using a glutathione S-transferase pull-down assay in which the cytoplasmic domain of CEACAM-4S was fused to glutathione S transferase, the fusion protein was incubated with cell lysates, and isolated proteins were sequenced by mass spectrometry. The interaction was confirmed first by reciprocal immunoprecipitations using anti-CEACAM1 and anti-annexin II antibodies and second by confocal laser microscopy showing co-localization of CEACAM1 with annexin II in mammary epithelial cells grown in Matrigel. In addition, CEACAM1 co-localized with p11, a component of the tetrameric AIIt complex at the plasma membrane, and with annexin II in secretory vesicles. Immobilized, oriented peptides from the cytoplasmic domain of CEACAM1-4S were shown to directly associate with bovine AIIt, which is 98% homologous to human AIIt, with average KD values of about 30 nM using surface plasmon resonance, demonstrating direct binding of functionally relevant AIIt to the cytoplasmic domain of CEACAM1-4S. PMID- 14522962 TI - Identification of human glutaminyl cyclase as a metalloenzyme. Potent inhibition by imidazole derivatives and heterocyclic chelators. AB - Human glutaminyl cyclase (QC) was identified as a metalloenzyme as suggested by the time-dependent inhibition by the heterocyclic chelators 1,10-phenanthroline and dipicolinic acid. The effect of EDTA on QC catalysis was negligible. Inactivated enzyme could be fully restored by the addition of Zn2+ in the presence of equimolar concentrations of EDTA. Little reactivation was observed with Co2+ and Mn2+. Other metal ions such as K+, Ca2+, and Ni2+ were inactive under the same conditions. Additionally, imidazole and imidazole derivatives were identified as competitive inhibitors of QC. An initial structure activity-based inhibitor screening of imidazole-derived compounds revealed potent inhibition of QC by imidazole N-1 derivatives. Subsequent data base screening led to the identification of two highly potent inhibitors, 3-[3-(1H-imidazol-1-yl)propyl]-2 thioxoimidazolidin-4-one and 1,4-bis-(imidazol-1-yl)-methyl-2,5-dimethylbenzene, which exhibited respective Ki values of 818 +/- 1 and 295 +/- 5 nm. The binding properties of the imidazole derivatives were further analyzed by the pH dependence of QC inhibition. The kinetically obtained pKa values of 6.94 +/- 0.02, 6.93 +/- 0.03, and 5.60 +/- 0.05 for imidazole, methylimidazole, and benzimidazole, respectively, match the values obtained by titrimetric pKa determination, indicating the requirement for an unprotonated nitrogen for binding to QC. Similarly, the pH dependence of the kinetic parameter Km for the QC-catalyzed conversion of H-Gln-7-ami-no-4-methylcoumarin also implies that only N-terminally unprotonated substrate molecules are bound to the active site of the enzyme, whereas turnover is not affected. The results reveal human QC as a metal dependent transferase, suggesting that the active site-bound metal is a potential site for interaction with novel, highly potent competitive inhibitors. PMID- 14522963 TI - Formation of acrolein-derived 2'-deoxyadenosine adduct in an iron-induced carcinogenesis model. AB - Acrolein is a representative carcinogenic aldehyde found ubiquitously in the environment and formed endogenously through oxidation reactions, such as lipid peroxidation and myeloperoxidase-catalyzed amino acid oxidation. It shows facile reactivity toward DNA to form an exocyclic DNA adduct. To verify the formation of acrolein-derived DNA adduct under oxidative stress in vivo, we raised a novel monoclonal antibody (mAb21) against the acrolein-modified DNA and found that the antibody most significantly recognized an acrolein-modified 2' -deoxyadenosine. On the basis of chemical and spectroscopic evidence, the major antigenic product of mAb21 was the 1,N6-propano-2' -deoxyadenosine adduct. The exposure of rat liver epithelial RL34 cells to acrolein resulted in a significant accumulation of the acrolein-2' -deoxyadenosine adduct in the nuclei. Formation of this adduct under oxidative stress in vivo was immunohistochemically examined in rats exposed to ferric nitrilotriacetate, a carcinogenic iron chelate that specifically induces oxidative stress in the kidneys of rodents. It was observed that the acrolein-2' -deoxyadenosine adduct was formed in the nuclei of the proximal tubular cells, the target cells of this carcinogenesis model. The same cells were stained with a monoclonal antibody 5F6 that recognizes an acrolein-lysine adduct, by which cytosolic accumulation of acrolein-modified proteins appeared. Similar results were also obtained from myeloperoxidase knockout mice exposed to the iron complex, suggesting that the myeloperoxidase-catalyzed oxidation system might not be essential for the generation of acrolein in this experimental animal carcinogenesis model. The data obtained in this study suggest that the formation of a carcinogenic aldehyde through lipid peroxidation may be causally involved in the pathophysiological effects associated with oxidative stress. PMID- 14522964 TI - Citrate oscillates in liver and pancreatic beta cell mitochondria and in INS-1 insulinoma cells. AB - Oscillations in citric acid cycle intermediates have never been previously reported in any type of cell. Here we show that adding pyruvate to isolated mitochondria from liver, pancreatic islets, and INS-1 insulinoma cells or adding glucose to intact INS-1 cells causes sustained oscillations in citrate levels. Other citric acid cycle intermediates measured either did not oscillate or possibly oscillated with a low amplitude. In INS-1 mitochondria citrate oscillations are in phase with NAD(P) oscillations, and in intact INS-1 cells citrate oscillations parallel oscillations in ATP, suggesting that these processes are co-regulated. Oscillations have been extensively studied in the pancreatic beta cell where oscillations in glycolysis, NAD(P)/NAD(P)H and ATP/ADP ratios, plasma membrane electrical activity, calcium levels, and insulin secretion have been well documented. Because the mitochondrion is the major site of ATP synthesis and NADH oxidation and the only site of citrate synthesis, mitochondria need to be synchronized for these factors to oscillate. In suspensions of mitochondria from various organs, most of the citrate is exported from the mitochondria. In addition, citrate inhibits its own synthesis. We propose that this enables citrate itself to act as one of the cellular messengers that synchronizes mitochondria. Furthermore, because citrate is a potent inhibitor of the glycolytic enzyme phosphofructokinase, the pacemaker of glycolytic oscillations, citrate may act as a metabolic link between mitochondria and glycolysis. Citrate oscillations may coordinate oscillations in mitochondrial energy production and anaplerosis with glycolytic oscillations, which in the beta cell are known to parallel oscillations in insulin secretion. PMID- 14522965 TI - Nick-dependent and -independent processing of large DNA loops in human cells. AB - DNA loop heterologies are products of normal DNA metabolism and can lead to severe genomic instability if unrepaired. To understand how human cells process DNA loop structures, a set of circular heteroduplexes containing a 30-nucleotide loop were constructed and tested for repair in vitro by human cell nuclear extracts. We demonstrate here that, in addition to the previously identified 5' nick-directed loop repair pathway (Littman, S. J., Fang, W. H., and Modrich, P. (1999) J. Biol. Chem. 274, 7474-7481), human cells can process large DNA loop heterologies in a loop-directed manner. The loop-directed repair specifically removes the loop structure and occurs only in the looped strand, and appears to require limited DNA synthesis. Like the nick-directed loop repair, the loop directed repair is independent of many known DNA repair pathways, including DNA mismatch repair and nucleotide excision repair. In addition, our data also suggest that an aphidicolin-sensitive DNA polymerase is involved in the excision step of the nick-directed loop repair pathway. PMID- 14522966 TI - Suppression of extracellular signal-related kinase and activation of p38 MAPK are two critical events leading to caspase-8- and mitochondria-mediated cell death in phytosphingosine-treated human cancer cells. AB - We previously demonstrated that the phytosphingosine-induced apoptosis was accompanied by the concomitant induction of both the caspase-8-mediated and mitochondrial activation-mediated apoptosis pathways. In the present study, we investigated the role of mitogen-activated protein kinases (MAPKs) in the activation of these two distinct cell death pathways induced by phytosphingosine in human cancer cells. Phytosphingosine caused strong induction of caspase-8 activity and caspase-independent Bax translocation to the mitochondria. A rapid decrease of phosphorylated ERK1/2 and a marked increase of p38 MAPK phosphorylation were observed within 10 min after phytosphingosine treatment. Activation of ERK1/2 by pretreatment with phorbol 12-myristate 13-acetate or forced expression of ERK1/2 attenuated phytosphingosine-induced caspase-8 activation. However, Bax translocation and caspase-9 activation was unaffected, indicating that down-regulation of the ERK activity is specifically required for the phytosphingosine-induced caspase-8-dependent cell death pathway. On the other hand, treatment with SB203580, a p38 MAPK-specific inhibitor, or expression of a dominant negative form of p38 MAPK suppressed phytosphingosine-induced translocation of the proapoptotic protein, Bax, from the cytosol to mitochondria, cytochrome c release, and subsequent caspase-9 activation but did not affect caspase-8 activation, indicating that activation of p38 MAPK is involved in the mitochondrial activation-mediated cell death pathway. Our results suggest that phytosphingosine can utilize two different MAPK signaling pathways for amplifying the apoptosis cascade, enhancing the understanding of the molecular mechanisms utilized by naturally occurring metabolites to regulate cell death. Molecular dissection of the signaling pathways that activate the apoptotic cell death machinery is critical for both our understanding of cell death events and development of cancer therapeutic agents. PMID- 14522967 TI - Identification of relaxin-3/INSL7 as a ligand for GPCR142. AB - We have recently identified the insulin-like peptide relaxin-3 (aka INSL7) as the endogenous ligand for an orphan G-protein-coupled receptor, GPCR135 (aka somatostatin- and angiotensin-like peptide receptor). Analysis of possible receptors related to GPCR135 revealed a single orphan receptor, GPCR142. Thus, we tested whether GPCR142 could also respond to relaxin-3 or related insulin-like molecules. Surprisingly, GPCR142 was activated by nanomolar concentrations of relaxin-3 but was completely unresponsive to all other known insulin-like peptides. We evaluated by reverse transcriptase-PCR the expression of GPCR142 mRNA in a variety of human tissues and found expression in brain, kidney, testis, thymus, placenta, prostate, salivary gland, thyroid, and colon. In an analysis of other species, we were able to find a full-length mouse homolog of GPCR142, but were unable to detect any complete GPCR142 transcripts in rat. With respect to intracellular signaling, GPCR142 is similar to GPCR135 in that it potently inhibits adenylate cyclase and stimulates 35S-GTPgammaS incorporation in response to relaxin-3. However, whereas GPCR135 signaling could be converted to calcium mobilization using a Gqi5 or Galpha16 G-proteins, GPCR142 was only capable of functioning in the presence of Galpha16. In the accompanying article (Liu, C., Eriste, E., Sutton, S., Chen, J., Roland, B., Kuei, C., Farmer, N., Jornvall, H., Sillard, R., and Lovenberg, T. W. (2003) J. Biol. Chem. 278, 50754-50764), we present the case that relaxin-3, which has previously been shown to bind to the relaxin receptor LGR7, is most likely the endogenous ligand for GPCR135. In this report, we show an additional receptor, GPCR142, which is also selectively activated by relaxin-3. However, the anatomical localization of GPCR142 suggests that GPCR142 may have different physiological functions. PMID- 14522969 TI - Reprogramming alternative pre-messenger RNA splicing through the use of protein binding antisense oligonucleotides. AB - Alternative pre-messenger RNA splicing is a major contributor to proteomic diversity in higher eukaryotes and represents a key step in the control of protein function in a large variety of biological systems. As a means of artificially altering splice site choice, we have investigated the impact of positioning proteins in the vicinity of 5' splice sites. We find that a recombinant GST-MS2 protein interferes with 5' splice site use, most efficiently when it binds upstream of that site. To broaden the use of proteins as steric inhibitors of splicing, we have tested the activity of antisense oligonucleotides carrying binding sites for the heterogeneous nuclear ribonucleoprotein A1/A2 proteins. In a HeLa cell extract, tailed oligonucleotides complementary to exonic sequences elicit strong shifts in 5' splice site selection. In four different human cell lines, an interfering oligonucleotide carrying A1/A2 binding sites also shifted the alternative splicing of the Bcl-x pre-mRNA more efficiently than oligonucleotides acting through duplex formation only. The use of protein-binding oligonucleotides that interfere with U1 small nuclear ribonucleoprotein binding therefore represents a novel and powerful approach to control splice site selection in cells. PMID- 14522968 TI - Identification of relaxin-3/INSL7 as an endogenous ligand for the orphan G protein-coupled receptor GPCR135. AB - GPCR135, publicly known as somatostatin- and angiotensin-like peptide receptor, is expressed in the central nervous system and its cognate ligand(s) has not been identified. We have found that both rat and porcine brain extracts stimulated 35S labeled guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) incorporation in cells over-expressing GPCR135. Multiple rounds of extraction, purification, followed by N-terminal sequence analysis of the ligand from porcine brain revealed that the ligand is a product of the recently identified gene, relaxin-3 (aka insulin-7 or INSL7). Recombinant human relaxin-3 potently stimulates GTPgammaS binding and inhibits cAMP accumulation in GPCR135 overexpressing cells with EC50 values of 0.25 and 0.35 nM, respectively. 125I-Relaxin-3 binds GPCR135 at high affinity with a Kd value of 0.31 nM. Relaxin-3 is the only member of the insulin/relaxin superfamily that can activate GPCR135. In situ hybridization showed that relaxin 3 mRNA is predominantly expressed in the dorsomedial ventral tegmental nucleus of the brainstem (aka nucleus incertus), as well as in discrete cells in the lateral periaqueductal gray and in the central gray nucleus. GPCR135 is expressed abundantly in the hypothalamus with discrete expression in the paraventricular nucleus of the hypothalamus and supraoptic nucleus, as well as in the cortex, septal nucleus, and preoptical area. Relaxin-3 has previously been shown to bind and activate the LGR7 relaxin receptor. However, we believe that neuroanatomical colocalization of GPCR135 and relaxin-3, coupled with a clear high affinity interaction, suggest that GPCR135 is the receptor for relaxin-3. The identification of relaxin-3 as the ligand for GPCR135 provides the framework for the discovery of a new brainstem/hypothalamus circuitry. PMID- 14522970 TI - Translocation of group 1 capsular polysaccharide in Escherichia coli serotype K30. Structural and functional analysis of the outer membrane lipoprotein Wza. AB - The late steps in assembly of capsular polysaccharides (CPS) and their translocation to the bacterial cell surface are not well understood. The Wza protein was shown previously to be required for the formation of the prototype group 1 capsule structure on the surface of Escherichia coli serotype K30 (Drummelsmith, J., and Whitfield, C. (2000) EMBO J. 19, 57-66). Wza is a conserved outer membrane lipoprotein that forms multimers adopting a ringlike structure, and collective evidence suggests a role for these structures in the export of capsular polymer across the outer membrane. Wza was purified in the native form and with a C-terminal hexahistidine tag. WzaHis6 was acylated and functional in capsule assembly, although its efficiency was slightly reduced in comparison to the native Wza protein. Ordered two-dimensional crystals of WzaHis6 were obtained after reconstitution of purified multimers into lipids. Electron microscopy of negatively stained crystals and Fourier filtering revealed ringlike multimers with an average outer diameter of 8.84 nm and an average central cavity diameter of 2.28 nm. Single particle analysis yielded projection structures at an estimated resolution of 3 nm, favoring a structure for the WzaHis6 containing eight identical subunits. A derivative of Wza (Wza*) in which the original signal sequence was replaced with that from OmpF showed that the native acylated N terminus of Wza is critical for formation of normal multimeric structures and for their competence for CPS assembly, but not for targeting Wza to the outer membrane. In the presence of Wza*, CPS accumulated in the periplasm but was not detected on the cell surface. Chemical cross-linking of intact cells suggested formation of a transmembrane complex minimally containing Wza and the inner membrane tyrosine autokinase Wzc. PMID- 14522972 TI - High extracellular Ca2+ hyperpolarizes human parathyroid cells via Ca(2+) activated K+ channels. AB - Membrane potential has a major influence on stimulus-secretion coupling in various excitable cells. The role of membrane potential in the regulation of parathyroid hormone secretion is not known. High K+-induced depolarization increases secretion from parathyroid cells. The paradox is that increased extracellular Ca2+, which inhibits secretion, has also been postulated to have a depolarizing effect. In this study, human parathyroid cells from parathyroid adenomas were used in patch clamp studies of K+ channels and membrane potential. Detailed characterization revealed two K+ channels that were strictly dependent of intracellular Ca2+ concentration. At high extracellular Ca2+, a large K+ current was seen, and the cells were hyperpolarized (-50.4 +/- 13.4 mV), whereas lowering of extracellular Ca2+ resulted in a dramatic decrease in K+ current and depolarization of the cells (-0.1 +/- 8.8 mV, p < 0.001). Changes in extracellular Ca2+ did not alter K+ currents when intracellular Ca2+ was clamped, indicating that K+ channels are activated by intracellular Ca2+. The results were concordant in cell-attached, perforated patch, whole-cell and excised membrane patch configurations. These results suggest that [Ca2+]o regulates membrane potential of human parathyroid cells via Ca2+-activated K+ channels and that the membrane potential may be of greater importance for the stimulus-secretion coupling than recognized previously. PMID- 14522971 TI - Identification of a novel glial cell line-derived neurotrophic factor-inducible gene required for renal branching morphogenesis. AB - In the developing kidney, activation of the rearrangement during transfection tyrosine kinase by glial cell line-derived neurotrophic factor (GDNF) is required for normal branching of the ureteric bud epithelium [corrected]. By differential display analysis we identified a novel GDNF-inducible gene (named GZF1) with a BTB/POZ (broad complex, tramtrack, and bric-a-brac)/(poxvirus and zinc finger) domain and 10 tandemly repeated zinc finger motifs. The up-regulation of the GZF1 gene showed two peaks at 1 h and 24-48 h after GDNF stimulation by Northern blotting. The late induction was also found at protein levels by Western blotting with anti-GZF1 antibody. As observed for other proteins with the BTB/POZ domain, the GZF1 protein had strong transcriptional repressive activity. Intriguingly, its expression was detected at high levels in branching ureteric buds and collecting ducts of mouse metanephric kidney in which RET was also expressed. Antisense phosphorothioated oligodeoxynucleotides of the GZF1 gene markedly impaired the ureteric bud branching in the metanephric organ culture, suggesting that the induction of GZF1 expression via the GDNF/RET signaling system is required for renal branching morphogenesis. PMID- 14522973 TI - Activation of the fatty acid alpha-dioxygenase pathway during bacterial infection of tobacco leaves. Formation of oxylipins protecting against cell death. AB - A pathogen-induced oxygenase showing homology to prostaglandin endoperoxide synthases-1 and -2 was recently characterized by in vitro experiments as a fatty acid alpha-dioxygenase catalyzing formation of unstable 2(R)-hydroperoxy fatty acids. To study the activity of this enzyme under in vivo conditions and to elucidate the fate of enzymatically produced 2-hydroperoxides, leaves of tobacco were analyzed for the presence of alpha-dioxygenase-generated compounds as well as for lipoxygenase (LOX) products and free fatty acids. Low basal levels of 2 hydroxylinolenic acid (0.4 nmol/g leaves fresh weight) and 8,11,14 heptadecatrienoic acid (0.1 nmol/g) could be demonstrated. These levels increased strongly upon infection with the bacterium Pseudomonas syringae pv syringae (548 and 47 nmol/g, respectively). Transgenic tobacco plants overexpressing alpha dioxygenase were developed, and incompatible infection of such plants led to a dramatic elevation of 2-hydroxylinolenic acid (1778 nmol/g) and 8,11,14 heptadecatrienoic acid (86 nmol/g), whereas the levels of LOX products were strongly decreased. Further analysis of oxylipins in infected leaves revealed the presence of a number of 2-hydroxy fatty acids differing with respect to chain length and degree of unsaturation as well as two new doubly oxygenated oxylipins identified as 2(R),9(S)-dihydroxy-10(E),12(Z),15(Z)-octadecatrienoic acid and 2(R),9(S)-dihydroxy-10(E),12(Z)-octadecadienoic acid. alpha-Dioxygenase-generated 2-hydroxylinolenic acid, and to a lesser extent lipoxygenase-generated 9 hydroxyoctadecatrienoic acid, exerted a tissue-protective effect in bacterially infected tobacco leaves. PMID- 14522974 TI - Methanethiosulfonate derivatives of rhodamine and verapamil activate human P glycoprotein at different sites. AB - The human multidrug resistance P-glycoprotein (P-gp, ABCB1) actively extrudes a broad range of potentially cytotoxic compounds out of the cell. Key steps in understanding the transport process are binding of drug substrates in the transmembrane domains, initiation of ATPase activity, and subsequent drug efflux. We used cysteine-scanning mutagenesis of the transmembrane segment residues and reaction with the thiol-reactive drug substrate analog of rhodamine, methane thiosulfonate-rhodamine (MTS-rhodamine), to test whether P-gp could be trapped in an activated state with high levels of ATPase activity. The presence of such an activated P-gp could be used to further investigate P-gp-drug substrate interactions. Single cysteine mutants (149) were treated with MTS-rhodamine, and ATPase activities were determined after removal of unreacted MTS-rhodamine. One mutant, F343C(TM6), showed a 5.8-fold increase in activity after reaction with MTS-rhodamine. Pre-treatment of mutant F343C with rhodamine B protected it from activation by MTS-rhodamine, indicating that residue Cys-343 contributes to the rhodamine-binding site. The ATPase activity of MTS-rhodamine-treated mutant F343C, however, was not stimulated further by colchicine or calcein-AM. By contrast, verapamil and Hoechst 33342 stimulated and inhibited, respectively, the ATPase activity of the MTS-rhodamine-treated mutant F343C. These results indicate that the MTS-rhodamine binding site overlaps that of colchicine and calcein-AM but not that of verapamil and Hoechst 33342 within the common drug-binding pocket. PMID- 14522975 TI - Crystal structure of the human centromere protein B (CENP-B) dimerization domain at 1.65-A resolution. AB - The human centromere protein B (CENP-B), a centromeric heterochromatin component, forms a homodimer that specifically binds to a distinct DNA sequence (the CENP-B box), which appears within every other alpha-satellite repeat. Previously, we determined the structure of the human CENP-B DNA-binding domain, CENP-B-(1-129), complexed with the CENP-B box DNA. In the present study, we determined the crystal structure of its dimerization domain (CENP-B-(540-599)), another functional domain of CENP-B, at 1.65-A resolution. CENP-B-(540-599) contains two alpha-helices, which are folded into an antiparallel configuration. The CENP-B (540-599) dimer formed a symmetrical, antiparallel, four-helix bundle structure with a large hydrophobic patch in which 23 residues of one monomer form van der Waals contacts with the other monomer. In the CENP-B-(540-599) dimer, the N terminal ends of CENP-B-(540-599) are oriented on opposite sides of the dimer. This CENP-B dimer configuration may be suitable for capturing two distant CENP-B boxes during centromeric heterochromatin formation. PMID- 14522976 TI - Defective potassium channel Kir2.1 trafficking underlies Andersen-Tawil syndrome. AB - Andersen-Tawil syndrome is a skeletal and cardiac muscle disease with developmental features caused by mutations in the inward rectifier K+ channel gene KCNJ2. Patients harboring these mutations exhibit extremely variable expressivities. To explore whether these mutations can be correlated with a specific patient phenotype, we expressed both wild-type (WT) and mutant genes cloned into a bi-cistronic vector. Functional expression in human embryonic kidney 293 cells showed that none of the mutant channels express current when present alone. When co-expressed with WT channels, only construct V302M-WT yields inward current. Confocal microscopy fluorescence revealed three patterns of channel expression in the cell: 1) mutations D71V, N216H, R218Q, and pore mutations co-assemble and co-localize to the membrane with the WT and exert a dominant-negative effect on the WT channels; 2) mutation V302M leads to channels that lose their ability to co-assemble with WT and traffic to the cell surface; 3) deletions Delta 95-98 and Delta 314-315 lead to channels that do not traffic to the membrane but retain their ability to co-assemble with WT channels. These data show that the Andersen-Tawil syndrome phenotype may occur through a dominant negative effect as well as through haplo-insufficiency and reveal amino acids critical in trafficking and conductance of the inward rectifier K+ channels. PMID- 14522977 TI - Single-walled carbon nanotubes are a new class of ion channel blockers. AB - Here we identify a novel class of biological membrane ion channel blockers called single-walled carbon nanotubes (SWNTs). SWNTs with diameter distributions peaked at approximately 0.9 and 1.3 nm, C60 fullerenes, multi wall nanotubes (MWNTs), and hyperfullerenes (nano-"onions") were synthesized by several techniques and applied to diverse channel types heterologously expressed in mammalian cells. External as-fabricated and purified SWNTs blocked K+ channel subunits in a dose dependent manner. Blockage was dependent on the shape and dimensions of the nanoparticles used and did not require any electrochemical interaction. SWNTs were more effective than the spherical fullerenes and, for both, diameter was the determining factor. These findings postulate new uses for SWNTs in biological applications and provide unexpected insights into the current view of mechanisms governing the interaction of ion channels with blocking molecules. PMID- 14522978 TI - Glutaredoxin exerts an antiapoptotic effect by regulating the redox state of Akt. AB - Glutaredoxin (GRX) is a small dithiol protein involved in various cellular functions, including the redox regulation of certain enzyme activities. GRX functions via a disulfide exchange reaction by utilizing the active site Cys-Pro Tyr-Cys. Here we demonstrated that overexpression of GRX protected cells from hydrogen peroxide (H2O2)-induced apoptosis by regulating the redox state of Akt. Akt was transiently phosphorylated, dephosphorylated, and then degraded in cardiac H9c2 cells undergoing H2O2-induced apoptosis. Under stress, Akt underwent disulfide bond formation between Cys-297 and Cys-311 and dephosphorylation in accordance with an increased association with protein phosphatase 2A. Overexpression of GRX protected Akt from H2O2-induced oxidation and suppressed recruitment of protein phosphatase 2A to Akt, resulting in a sustained phosphorylation of Akt and inhibition of apoptosis. This effect was reversed by cadmium, an inhibitor of GRX. Furthermore an in vitro assay revealed that GRX reduced oxidized Akt in concert with glutathione, NADPH, and glutathione disulfide reductase. Thus, GRX plays an important role in protecting cells from apoptosis by regulating the redox state of Akt. PMID- 14522979 TI - Regulation of inducible heparanase gene transcription in activated T cells by early growth response 1. AB - Cleavage of heparan sulfate by the beta-D-endoglucuronidase heparanase (HPSE) is a fundamental event in a number of important physiological processes including inflammation, wound healing, and angiogenesis. HPSE activity has also been directly correlated with pathological conditions such as tumor growth and metastasis and autoimmune disease. The tight regulation of HPSE expression and function is critical to ensure homeostasis of the normal physiological processes to which it contributes and to prevent imbalance toward pathological situations. Little is known about the transcriptional mechanisms that regulate HPSE expression. In this study we have shown human HPSE gene transcription in Jurkat T cells is induced upon activation. Functional analysis of the HPSE promoter has identified a 280-bp region that is highly inducible. Mutation studies together with supershift experiments have identified a 4-bp motif that binds the transcription factor early growth response-1 (Egr1) and is critical in regulating inducible HPSE gene transcription. Furthermore, the overexpression of Egr1 resulted in the enhanced activation of the HPSE promoter. By using MAPK pathway inhibitors, we have also shown that inducible expression of HPSE mRNA and the activity of the 280-bp HPSE promoter element are dependent on the ERK1/2 (MEK1/2) pathway. This pathway is critical for induction of Egr1 expression at both the mRNA and protein level in T cells, an observation that provides further support to Egr1 playing an important role as a key activator of HPSE expression. In addition, HPSE and Egr1 were shown to co-localize by immunohistochemistry to invading mononuclear leukocytes in actively induced experimental autoimmune encephalomyelitis in rats. These findings provide the first insight into the mechanisms controlling inducible transcription of the HPSE gene, and could represent an important lead into understanding how HPSE expression is deregulated in metastatic tumor cells. PMID- 14522980 TI - Identification and characterization of GCP16, a novel acylated Golgi protein that interacts with GCP170. AB - GCP170, a member of the golgin family associated with the cytoplasmic face of the Golgi membrane, was found to have a Golgi localization signal at the NH2-terminal region (positions 137-237). Using this domain as bait in the yeast two-hybrid screening system, we identified a novel protein that interacted with GCP170. The 2.0-kilobase mRNA encoding a 137-amino acid protein of 16 kDa designated GCP16 was ubiquitously expressed. Immunofluorescence microscopy showed that GCP16 was co-localized with GCP170 and giantin in the Golgi region. Despite the absence of a hydrophobic domain sufficient for participating in membrane localization, GCP16 was found to be tightly associated with membranes like an integral membrane protein. Labeling experiments with [3H]palmitic acid and mutational analysis demonstrated that GCP16 was acylated at Cys69 and Cys72, accounting for its tight association with the membrane. A mutant without potential acylation sites (C69A/C72A) was no longer localized to the Golgi, indicating that the acylation is prerequisite for the Golgi localization of GCP16. Although the mutant GCP16, even when overexpressed, had no effect on protein transport, overexpression of the wild type GCP16 caused an inhibitory effect on protein transport from the Golgi to the cell surface. Taken together, these results indicate that GCP16 is the acylated membrane protein, associated with GCP170, and possibly involved in vesicular transport from the Golgi to the cell surface. PMID- 14522981 TI - In vitro and in vivo dimerization of human endonuclease III stimulates its activity. AB - Human endonuclease III (hNTH1), a DNA glycosylase with associated abasic lyase activity, repairs various mutagenic and toxic-oxidized DNA lesions, including thymine glycol. We demonstrate for the first time that the full-length hNTH1 positively cooperates in product formation as a function of enzyme concentration. The protein concentrations that caused cooperativity in turnover also exhibited dimerization, independent of DNA binding. Earlier we had found that the hNTH1 consists of two domains: a well conserved catalytic domain, and an inhibitory N terminal tail. The N-terminal truncated proteins neither undergo dimerization, nor do they show cooperativity in turnover, indicating that the homodimerization of hNTH1 is specific and requires the N-terminal tail. Further kinetic analysis at transition states reveals that this homodimerization stimulates an 11-fold increase in the rate of release of the final product, an AP-site with a 3'-nick, and that it does not affect other intermediate reaction rates, including those of DNA N-glycosylase or AP lyase activities that are modulated by previously reported interacting proteins, YB-1, APE1, and XPG. Thus, the site of modulating action of the dimer on the hNTH1 reaction steps is unique. Moreover, the high intranuclear (2.3 microM) and cytosolic (0.65 microM) concentrations of hNTH1 determined here support the possibility of in vivo dimerization; indeed, in vivo protein cross-linking showed the presence of the dimer in the nucleus of HeLa cells. Therefore, it is likely that the dimerization of hNTH1 involving the N terminal tail masks the inhibitory effect of this tail and plays a critical role in its catalytic turnover in the cell. PMID- 14522983 TI - Molecular identification of Aggrus/T1alpha as a platelet aggregation-inducing factor expressed in colorectal tumors. AB - Platelets play an important role in hemostasis, thrombosis, and antimicrobial host defense and are also involved in the induction of inflammation, tissue repair, and tumor metastasis. We have previously characterized the platelet aggregation-inducing sialoglycoprotein (Aggrus/gp44) overexpressed on the surface of tumor cells. Because a platelet aggregation-neutralizing 8F11 monoclonal antibody that could specifically recognize Aggrus suppressed tumor-induced platelet aggregation, we have previously purified Aggrus by 8F11-affinity chromatography and found that purified Aggrus possessed the ability to induce aggregation of platelets. Here we show that Aggrus is identical to the T1alpha/gp38P/OTS-8 antigen, the function of which in tumors is unknown. Expression of mouse Aggrus and its human homologue (also known as T1alpha-2/gp36) induced platelet aggregation without requiring plasma components. Using the 8F11 antibody, we identified the highly conserved platelet aggregation-stimulating domain with putative O-glycosylated threonine residues as the critical determinant for exhibiting platelet aggregation-inducing capabilities. We compared the expression level of human aggrus mRNA using an array containing 160 cDNA pair samples derived from multiple human tumorigenic and corresponding normal tissues from individual patients. We found that expression level of aggrus was enhanced in most colorectal tumor patients. To confirm the protein expression, we generated anti-human Aggrus polyclonal antibodies. Immunohistochemical analysis revealed that Aggrus expression was frequently up regulated in colorectal tumors. These results suggest that Aggrus/T1alpha is a newly identified, platelet aggregation-inducing factor expressed in colorectal tumors. PMID- 14522982 TI - Presence of prepackaged mRNA in virions of DNA adenovirus. AB - Ran GTPase has been shown to be involved in host innate immune response, and two alleles, RanT/n and RanC/d, which differ from each other by a single nucleotide, have opposite effects on host innate immune response. In this study, we showed that although intravenous administration in mice with either Ran cDNA using an identical adenovirus (Ad) vector resulted in no significant difference in vector tissue distribution, intraperitoneal administration resulted in effective vector transduction into peritoneal macrophages, coupled with a striking difference in vector tissue distribution in 2 h or less. We further demonstrated the presence of prepackaged RNA in virions of Ad vectors, in cells actively producing Ad virus particles, and in cells very shortly after Ad infection. Real-time PCR analysis confirmed the presence of prepackaged RNA and estimated the copy number to be one per viral genome. The prepackaged viral mRNA could be used for translation into proteins, as shown by experiments in which the transcriptional inhibitor actinomycin-D was used. Hence, translation of Ran proteins from prepackaged viral mRNA immediately after virus uncoating in the cytoplasm is one mechanism that would account for an early difference in Ad-vector tissue distribution after efficient gene transfer into macrophages. PMID- 14522984 TI - The nature of the rate-limiting steps in the refolding of the cofactor-dependent protein aspartate aminotransferase. AB - The refolding of mitochondrial aspartate aminotransferase (mAAT; EC 2.6.1.1) has been studied following unfolding in 6 m guanidine hydrochloride for different periods of time. Whereas reactivation of equilibrium-unfolded mAAT is sigmoidal, reactivation of the short term unfolded protein displays a double exponential behavior consistent with the presence of fast and slow refolding species. The amplitude of the fast phase decreases with increasing unfolding times (k approximately 0.75 min(-1) at 20 degrees C) and becomes undetectable at equilibrium unfolding. According to hydrogen exchange and stopped-flow intrinsic fluorescence data, unfolding of mAAT appears to be complete in less than 10 s, but hydrolysis of the Schiff base linking the coenzyme pyridoxal 5'-phosphate (PLP) to the polypeptide is much slower (k approximately 0.08 min(-1)). This implies the existence in short term unfolded samples of unfolded species with PLP still attached. However, since the disappearance of the fast refolding phase is about 10-fold faster than the release of PLP, the fast refolding phase does not correspond to folding of the coenzyme-containing molecules. The fast refolding phase disappears more rapidly in the pyridoxamine and apoenzyme forms of mAAT, both of which lack covalently attached cofactor. Thus, bound PLP increases the kinetic stability of the fast refolding unfolding intermediates. Conversion between fast and slow folding forms also takes place in an early folding intermediate. The presence of cyclophilin has no effect on the reactivation of either equilibrium or short term unfolded mAAT. These results suggest that proline isomerization may not be the only factor determining the slow refolding of this cofactor-dependent protein. PMID- 14522985 TI - Dominant negative Rab3D mutants reduce GTP-bound endogenous Rab3D in pancreatic acini. AB - Two dominant negative mutants of Rab3D, N135I and T36N were recently reported to inhibit the early phase of regulated amylase secretion from mouse pancreatic acini (Chen, X., Edwards, J. A., Logsdon, C. D., Ernst, S. A., and Williams, J. A. (2002) J. Biol. Chem. 277, 18002-18009). Immunocytochemical studies showed that while the wild-type Rab3D localized to zymogen granules, the two dominant negative mutants did not localize to granules and were primarily in the basolateral regions of the cell. The present study, therefore, evaluated the potential mechanisms by which the dominant negative mutants might act. An affinity precipitation assay based on the property of the Rab3 effector Rim1 to interact only with GTP-bound Rab3D was developed. 78.9 +/- 4.5% of total endogenous Rab3D was found in the GTP-bound form. Overexpression of HA-tagged Rab3D, and its Q81L, N135I, and T36N mutants had no effect on the total amount of endogenous Rab3D. However, the dominant negative mutants, T36N and N135I, reduced GTP-bound endogenous Rab3D by 70.0 +/- 3.5% and 72.7 +/- 1.2%, respectively, while the wild-type Rab3D and Q81L mutant had no effect. Triton X-114 phase separation and cell fractionation studies showed that dominant negative Rab3D mutants did not alter isoprenylation or membrane association of endogenous Rab3D. The dominant negative Rab3D did not affect the amount of endogenous Rab3D on purified zymogen granules as assessed by either Western blotting or immunocytochemistry, but reduced the GTP-bound form by 78.6 +/- 3.3%. The two dominant negative Rab3D mutants, therefore, interfere with endogenous Rab3D function by blocking the GDP/GTP exchange but not zymogen granule targeting of endogenous Rab3D. PMID- 14522986 TI - Yeast xrs2 binds DNA and helps target rad50 and mre11 to DNA ends. AB - Saccharomyces cerevisiae Rad50, Mre11, and Xrs2 proteins are involved in homologous recombination, non-homologous end-joining, DNA damage checkpoint signaling, and telomere maintenance. These proteins form a stable complex that has nuclease, DNA binding, and DNA end recognition activities. Of the components of the Rad50.Mre11.Xrs2 complex, Xrs2 is the least characterized. The available evidence is consistent with the idea that Xrs2 recruits other protein factors in reactions that pertain to the biological functions of the Rad50.Mre11.Xrs2 complex. Here we present biochemical evidence that Xrs2 has an associated DNA binding activity that is specific for DNA structures. We also define the contributions of Xrs2 to the activities of the Rad50.Mre11.Xrs2 complex. Importantly, we demonstrate that Xrs2 is critical for targeting of Rad50 and Mre11 to DNA ends. Thus, Xrs2 likely plays a direct role in the engagement of DNA substrates by the Rad50. Mre11.Xrs2 complex in various biological processes. PMID- 14522987 TI - The amino acid sequence 442GDASE446 in Na/K-ATPase is an important motif in forming the high and low affinity ATP binding pockets. AB - A highly conserved amino acid sequence 442GDASE446 in the ATP binding pocket of rat Na/K-ATPase was mutated, and the resulting proteins, G442A, G442P, D443A, S445A, and E446A, were expressed in HeLa cells to investigate the effect of individual ligands on Na/K-ATPase. The apparent Km for the high and low affinity ATP effects was estimated by ATP concentration dependence for the formation of the Na-dependent phosphoenzyme (Kmh) and Na/K-ATPase activity (Kml). The apparent Km for p-nitrophenylphosphate (pNPP) for K-dependent-pNPPase (KmP) and its inhibition by ATP (Ki,0.5) and the apparent Km for Mg2+, Na+, K+, and vanadate in Na/K-ATPase were also estimated. For all the mutants, the value for ATP was approximately 2-10-fold larger than that of the wild type. While the turnover number for Na/K-ATPase activity were unaffected or reduced by 20 approximately 50% in mutants G442(A/P) and D443A. Although both affinities for ATP effects were reduced as a result of the mutations, the ratio, Kml Kmh, for each mutant was 1.3 approximately 3.7, indicating that these mutations had a greater impact on the low affinity ATP effect than on the high affinity effect. Each KmP value with the turnover number suggests that these mutations favor the binding of pNPP over that of ATP. These data and others indicate that the sequence 442GDASE446 in the ATP binding pocket is an important motif that it is involved in both the high and low affinity ATP effects rather than in free Mg2+, Na+, and K+ effects. PMID- 14522989 TI - Yeast RNA polymerase II lacking the Rpb9 subunit is impaired for interaction with transcription factor IIF. AB - Previous studies have shown that transcription factors IIB (TFIIB), IIF (TFIIF), and RNA polymerase II (RNAPII) play important roles in determining the position of mRNA 5'-ends in the yeast Saccharomyces cerevisiae. Yeast strains containing a deletion of the small, nonessential Rpb9 subunit of RNAPII exhibit an upstream shift in the positions of mRNA 5'-ends, whereas mutation of the large subunit of yeast TFIIF (Tfg1) can suppress downstream shifts that are conferred by mutations in TFIIB. In this study, we report an approach for the production of functional recombinant yeast holo-TFIIF (Tfg1-Tfg2 complex) and use of the recombinant protein in both reconstituted transcription assays and gel mobility shifts in order to investigate the biochemical alterations associated with the deltaRpb9 polymerase. The results demonstrated that upstream shifts in the positions of mRNA 5'-ends could be conferred by the deltaRpb9 RNAPII in transcription reactions reconstituted with highly purified yeast general transcription factors and, importantly, that these shifts are associated with an impaired interaction between the DeltaRpb9 polymerase and TFIIF. Potential mechanisms by which an altered interaction between the DeltaRpb9 RNAPII and TFIIF confers an upstream shift in the positions of mRNA 5'-ends are discussed. PMID- 14522988 TI - PGC-1alpha activates CYP7A1 and bile acid biosynthesis. AB - Cholesterol 7-alpha-hydroxylase (CYP7A1) is the key enzyme that commits cholesterol to the neutral bile acid biosynthesis pathway and is highly regulated. In the current studies, we have uncovered a role for the transcriptional co-activator PGC-1alpha in CYP7A1 gene transcription. PGC-1alpha plays a vital role in adaptive thermogenesis in brown adipose tissue and stimulates genes important to mitochondrial function and oxidative metabolism. It is also involved in the activation of hepatic gluconeogenesic gene expression during fasting. Because the mRNA for CYP7A1 was also induced in mouse liver by fasting, we reasoned that PGC-1alpha might be an important co-activator for CYP7A1. Here we show that PGC-1alpha and CYP7A1 are also co-induced in livers of mice in response to streptozotocin induced diabetes. Additionally, infection of cultured HepG2 cells with a recombinant adenovirus expressing PGC-1alpha directly activates CYP7A1 gene expression and increases bile acid biosynthesis as well. Furthermore, we show that PGC-1alpha activates the CYP7A1 promoter directly in transient transfection assays in cultured cells. Thus, PGC-1alpha is a key activator of CYP7A1 and bile acid biosynthesis and is likely responsible for the fasting and diabetes dependent induction of CYP7A1. PGC-1alpha has already been shown to be a critical activator of several other oxidative processes including adaptive thermogenesis and fatty acid oxidation. Our studies provide further evidence of the fundamental role played by PGC-1alpha in oxidative metabolism and define PGC-1alpha as a link between diabetes and bile acid metabolism. PMID- 14522990 TI - Repair of oxidized bases in DNA bubble structures by human DNA glycosylases NEIL1 and NEIL2. AB - Repair of oxidatively damaged bases in the genome via the base excision repair pathway is initiated with excision of these lesions by DNA glycosylases with broad substrate range. The newly discovered human DNA glycosylases, NEIL1 and NEIL2, are distinct in structural features and reaction mechanism from the previously characterized NTH1 and OGG1 but act on many of the same substrates. However, NEIL2 shows a unique preference for excising lesions from a DNA bubble, whereas NTH1 and OGG1 are only active with duplex DNA. NEIL1 also excises efficiently 5-hydroxyuracil, an oxidation product of cytosine, from the bubble and single-stranded DNA but does not have strong activity toward 8-oxoguanine in the bubble. The dichotomy in the activity of NEILs versus NTH1/OGG1 for bubble versus duplex DNA substrates is consistent with higher affinity of the NEILs for the bubble structures of both damaged and undamaged DNA relative to duplex structure. These observations suggest that the NEILs are functionally distinct from OGG1/NTH1 in vivo. OGG1/NTH1-independent repair of oxidized bases in the transcribed sequences supports the possibility that NEILs are preferentially involved in repair of lesions in DNA bubbles generated during transcription and/or replication. PMID- 14522991 TI - Direct observation of G-protein binding to the human delta-opioid receptor using plasmon-waveguide resonance spectroscopy. AB - Using a recently developed method (Salamon, Z., Macleod, H. A., and Tollin, G. (1997) Biophys. J. 73, 2791-2797), plasmon-waveguide resonance spectroscopy, we have been able, for the first time, to directly measure the binding between the human brain delta-opioid receptor (hDOR) and its G-protein effectors in real time. We have found that the affinity of the G-proteins toward the receptor is highly dependent on the nature of the ligand pre-bound to the receptor. The highest affinity was observed when the receptor was bound to an agonist ( approximately 10 nm); the lowest when receptor was bound to an antagonist ( approximately 500 nm); and no binding at all was observed when the receptor was bound to an inverse agonist. We also have found direct evidence for the existence of an additional G-protein binding conformational state that corresponds to the unliganded receptor, which has a G-protein binding affinity of approximately 60 nm. Furthermore, GTP binding to the receptor.G-protein complex was only observed when the agonist was pre-bound. Similar studies were carried out using the individual G-protein subtypes for both the agonist and the unliganded receptor. Significant selectivity toward the different G-protein subtypes was observed. Thus, the unliganded receptor had highest affinity toward the Galphao (Kd approximately 20 nm) and lowest affinity toward the Galphai2 ( approximately 590 nm) subtypes, whereas the agonist-bound state had highest affinity for the Galphao and Galphai2 subtypes (Kd approximately 9 nm and approximately 7 nm, respectively). GTP binding was also highly selective, both with respect to ligand and G-protein subtype. We believe that this methodology provides a powerful new way of investigating transmembrane signaling. PMID- 14522992 TI - Characterization of the interaction of a recombinant soluble neuroligin-1 with neurexin-1beta. AB - Neuroligins, proteins of the alpha/beta-hydrolase fold family, are found as postsynaptic transmembrane proteins whose extracellular domain associates with presynaptic partners, proteins of the neurexin family. To characterize the molecular basis of neuroligin interaction with neurexin-beta, we expressed five soluble and exportable forms of neuroligin-1 from recombinant DNA sources, by truncating the protein before the transmembrane span near its carboxyl terminus. The extracellular domain of functional neuroligin-1 associates as a dimer when analyzed by sedimentation equilibrium. By surface plasmon resonance, we established that soluble neuroligins-1 bind neurexin-1beta, but the homologous alpha/beta-hydrolase fold protein, acetylcholinesterase, failed to associate with the neurexins. Neuroligin-1 has a unique N-linked glycosylation pattern in the neuroligin family, and glycosylation and its processing modify neuroligin activity. Incomplete processing of the protein and enzymatic removal of the oligosaccharides chain or the terminal sialic acids from neuroligin-1 enhance its activity, whereas deglycosylation of neurexin-1beta did not alter its association capacity. In particular, the N-linked glycosylation at position 303 appears to be a major determinant in modifying the association with neurexin-1beta. We show here that glycosylation processing of neuroligin, in addition to mRNA splicing and gene selection, contributes to the specificity of the neurexin beta/neuroligin-1 association. PMID- 14522993 TI - Isoform-specific regulation of insulin-dependent glucose uptake by Akt/protein kinase B. AB - Recent data have implicated the serine/threonine protein kinase Akt/protein kinase B (PKB) in a diverse array of physiological pathways, raising the question of how biological specificity is maintained. Partial clarification derived from the observation that mice deficient in either of the two isoforms, Akt1/PKBalpha or Akt2/PKBbeta, demonstrate distinct abnormalities, i.e. reduced organismal size or insulin resistance, respectively. However, the question still persists as to whether these divergent phenotypes are due exclusively to tissue-specific differences in isoform expression or distinct capacities for signaling intrinsic to the two proteins. Here we show that Akt2/PKBbeta-/- adipocytes derived from immortalized mouse embryo fibroblasts display significantly reduced insulin stimulated hexose uptake, clearly establishing that the partial defect in glucose disposal in these mice derives from lack of a cell autonomous function of Akt2/PKBbeta. Moreover, in adipocytes differentiated from primary fibroblasts or immortalized mouse embryo fibroblasts, and brown preadipocytes the absence of Akt2/PKBbeta resulted in reduction of insulin-induced hexose uptake and glucose transporter 4 (GLUT4) translocation, whereas Akt1/PKBalpha was dispensable for this effect. Most importantly, hexose uptake and GLUT4 translocation were completely restored after re-expression of Akt2/PKBbeta in Akt2/PKBbeta-/- adipocytes, but overexpression of Akt1/PKBalpha at comparable levels was ineffective at rescuing insulin action to normal. These results show that the Akt1/PKBalpha and Akt2/PKBbeta isoforms are uniquely adapted to preferentially transmit distinct biological signals, and this property is likely to contribute significantly to the ability of Akt/PKB to play a role in diverse processes. PMID- 14522994 TI - SHP-2 regulates SOCS-1-mediated Janus kinase-2 ubiquitination/degradation downstream of the prolactin receptor. AB - The protein tyrosine phosphatase SHP-2 is an important regulator of the Janus kinase-2 (Jak2)/signal transducer and activator of transcription (Stat) pathway downstream of the cytokine/prolactin receptor family. We report that SHP-2 dephosphorylates tyrosine (Tyr-1007) of Jak2 kinase, a critical recruitment site for the ubiquitin ligase-associated inhibitory protein suppressor of cytokine signaling-1 (SOCS-1), thereby contributing to Jak2 stability. Inactivation of SHP 2 function by blocking receptor/SHP-2 association or by using a catalytically inactive mutant of SHP-2 led to a marked increase in Jak2 ubiquitination/degradation, Jak2 phosphorylation on Tyr-1007, and Jak2/SOCS-1 association. Furthermore, functional studies indicate that modulating the interaction of Jak2/SOCS-1 by SHP-2 is essential for prolactin/Stat5-mediated signaling. Together our results provide a novel function for SHP-2 as a positive regulator of cytokine receptor signaling by regulating ubiquitination/degradation pathways. PMID- 14522995 TI - Separate roles for the Golgi apparatus and lysosomes in the sequestration of drugs in the multidrug-resistant human leukemic cell line HL-60. AB - The sequestration of drugs away from cellular target sites into cytoplasmic organelles of multidrug-resistant (MDR) cancer cells has been recently shown to be a cause for ineffective drug therapy. This process is poorly understood despite the fact that it has been observed in a large number of MDR cancer cell lines. Analysis of drug sequestration in these cells has traditionally been done using fluorescent anthracycline antibiotics (i.e. daunorubicin, doxorubicin). This narrow selection of substrates has resulted in a limited understanding of sequestration mechanisms and the intracellular compartments that are involved. To better characterize this phenotype, we chose to examine the sequestration of molecules having different acid/base properties in the MDR HL-60 human leukemic cell line. Here we show that weakly basic drug daunorubicin is sequestered into lysosomes according to a pH partitioning type mechanism, whereas sulforhodamime 101, a zwitterionic molecule, is sequestered into the Golgi apparatus through a drug transporter-mediated process. Quantitative intracellular pH measurements reveal that the lysosome-tocytosol pH gradient is expanded in the MDR line. Moreover, the MDR cells overexpress the multidrug resistance-related protein (MRP1), which is localized to the Golgi apparatus. These results demonstrate, for the first time, that two distinct mechanisms for intracellular compartmentalization are operational in a single MDR cell line. PMID- 14522996 TI - Mechanism of nicotinamide inhibition and transglycosidation by Sir2 histone/protein deacetylases. AB - Silent information regulator 2 (Sir2) enzymes catalyze NAD+-dependent protein/histone deacetylation, where the acetyl group from the lysine epsilon amino group is transferred to the ADP-ribose moiety of NAD+, producing nicotinamide and the novel metabolite O-acetyl-ADP-ribose. Sir2 proteins have been shown to regulate gene silencing, metabolic enzymes, and life span. Recently, nicotinamide has been implicated as a direct negative regulator of cellular Sir2 function; however, the mechanism of nicotinamide inhibition was not established. Sir2 enzymes are multifunctional in that the deacetylase reaction involves the cleavage of the nicotinamide-ribosyl, cleavage of an amide bond, and transfer of the acetyl group ultimately to the 2'-ribose hydroxyl of ADP-ribose. Here we demonstrate that nicotinamide inhibition is the result of nicotinamide intercepting an ADP-ribosyl-enzyme-acetyl peptide intermediate with regeneration of NAD+ (transglycosidation). The cellular implications are discussed. A variety of 3-substituted pyridines was found to be substrates for enzyme-catalyzed transglycosidation. A Bronsted plot of the data yielded a slope of +0.98, consistent with the development of a nearly full positive charge in the transition state, and with basicity of the attacking nucleophile as a strong predictor of reactivity. NAD+ analogues including beta-2'-deoxy-2'-fluororibo NAD+ and a His-to-Ala mutant were used to probe the mechanism of nicotinamide ribosyl cleavage and acetyl group transfer. We demonstrate that nicotinamide ribosyl cleavage is distinct from acetyl group transfer to the 2'-OH ribose. The observed enzyme-catalyzed formation of a labile 1'-acetylated-ADP-fluororibose intermediate using beta-2'-deoxy-2'-fluororibo-NAD+ supports a mechanism where, after nicotinamide-ribosyl cleavage, the carbonyl oxygen of acetylated substrate attacks the C-1' ribose to form an initial iminium adduct. PMID- 14522997 TI - Constitutive signaling of the human cytomegalovirus-encoded receptor UL33 differs from that of its rat cytomegalovirus homolog R33 by promiscuous activation of G proteins of the Gq, Gi, and Gs classes. AB - The human cytomegalovirus (HCMV) UL33 gene is conserved among all beta herpesviruses and encodes a protein that shows sequence similarity with chemokine receptors belonging to the family of G protein-coupled receptors. Here, we show that HCMV UL33 is predominantly transcribed as a spliced mRNA of which the 5' terminus is localized 55 bp upstream of the start codon. Like its homolog from rat cytomegalovirus (RCMV), R33, UL33 activates multiple signaling pathways in a ligand-independent manner. Although both receptors constitutively activate phospholipase C via G(q/11), and partially via G(i/o)-mediated pathways, they exhibit profound differences in the modulation of cAMP-responsive element (CRE) activation. R33 constitutively inhibits, whereas UL33 constitutively enhances CRE mediated transcription. For R33, the inhibition of CRE-driven transcription is entirely G(i/o)-mediated. For UL33, however, CRE-mediated transcription is modulated not only through coupling to Galpha(i/o) but also through coupling to Galphas. In addition, UL33 was found to enhance CRE activation through the Rho/p38 pathway, via Gbetagamma. Interestingly, by studying chimeric UL33/R33 proteins, we found the C-terminal cytoplasmic tail of UL33, but not that of R33, to be responsible for the activation of G(i/o) proteins. A UL33-deficient variant of HCMV was generated to analyze UL33-signaling properties in a physiologically relevant model system. Data obtained with infected cells show that HCMV induces CRE activation, and this effect is, at least in part, dependent on UL33 expression. Taken together, our data indicate that constitutive signaling of UL33 differs from that of R33 by promiscuous activation of G proteins of the Gq, G(i/o), as well as Gs class. Thus, HCMV may effectively use UL33 to orchestrate multiple signaling networks within infected cells. PMID- 14522998 TI - Use of proteomics to demonstrate a hierarchical oxidative stress response to diesel exhaust particle chemicals in a macrophage cell line. AB - Epidemiological studies demonstrate an association between short term exposure to ambient particulate matter (PM) and cardiorespiratory morbidity and mortality. Although the biological mechanisms of these adverse effects are unknown, emerging data suggest a key role for oxidative stress. Ambient PM and diesel exhaust particles (DEP) contain redox cycling organic chemicals that induce pro-oxidative and pro-inflammatory effects in the lung. These responses are suppressed by N acetylcysteine (NAC), which directly complexes to electrophilic DEP chemicals and exert additional antioxidant effects at the cellular level. A proteomics approach was used to study DEP-induced responses in the macrophage cell line, RAW 264.7. We demonstrate that in the dose range 10-100 microg/ml, organic DEP extracts induce a progressive decline in the cellular GSH/GSSG ratio, in parallel with a linear increase in newly expressed proteins on the two-dimensional gel. Using matrix-assisted laser desorption ionization time-of-flight mass spectrometry and electrospray ionization-liquid chromatography/mass spectrometry/mass spectrometry analysis, 32 newly induced/NAC-suppressed proteins were identified. These include antioxidant enzymes (e.g. heme oxygenase-1 and catalase), pro-inflammatory components (e.g. p38MAPK and Rel A), and products of intermediary metabolism that are regulated by oxidative stress. Heme oxygenase-1 was induced at low extract dose and with minimal decline in the GSH/GSSG ratio, whereas MAP kinase activation required a higher chemical dose and incremental levels of oxidative stress. Moreover, at extract doses >50 microg/ml, there is a steep decline in cellular viability. These data suggest that DEP induce a hierarchical oxidative stress response in which some of these proteins may serve as markers for oxidative stress during PM exposures. PMID- 14522999 TI - Interaction with a membrane surface triggers a reversible conformational change in Bax normally associated with induction of apoptosis. AB - The Bcl-2 family member Bax is an apoptosis-promoting protein that normally resides in an inactive state within the cytoplasm of healthy cells. Upon induction of apoptosis by diverse stimuli, Bax undergoes a conformational change and translocates to mitochondria, where it oligomerizes and forms pores that allow the release of cytochrome c and other cytotoxic factors. Protein-protein interactions between Bax and other Bcl-2 family members are strongly implicated in Bax activation, but a compelling case has recently been made for the involvement of lipids in this process as well. Here we report that purified Bax undergoes a reversible conformational change upon incubation with lipid vesicles in the absence of other proteins. This Bax-liposome interaction does not depend on a specific lipid composition. Changes in Bax conformation were observed by immunoprecipitation with the conformation-specific antibody 6A7, circular dichroism spectroscopy, and differential scanning calorimetry. Although liposomes induced Bax to become 6A7-reactive (a feature normally associated with the onset of apoptosis), the protein did not insert into membranes, become oligomeric, or form pores, clearly indicating that other triggers are required for Bax to achieve its final pro-apoptotic state. Indeed, the lipid-induced Bax conformational change is shown to be required for tBid-induced Bax oligomerization and pore formation, putting it upstream of tBid activity in this molecular pathway to Bax activation. These data demonstrate that Bax is sensitized to activation by transient interaction with lipid membrane surfaces and provide evidence that Bax activation proceeds in a stepwise fashion, with multiple triggers and potential levels of regulation. PMID- 14523000 TI - Aurora-A kinase maintains the fidelity of early and late mitotic events in HeLa cells. AB - Aurora-A, a member of the Aurora/Ipl1-related kinase family, is overexpressed in various types of cancer and considered to play critical roles in tumorigenesis. To better understand the pathological effect of Aurora-A activation, it is first necessary to elucidate the physiological functions of Aurora-A. Here, we have investigated the roles of Aurora-A in mitotic progression with the small interfering RNA, antibody microinjection, and time lapse microscopy using human cells. We demonstrated that suppression of Aurora-A by small interfering RNA caused multiple events to fail in mitosis, such as incorrect separation of centriole pairs, misalignment of chromosomes on the metaphase plate, and incomplete cytokinesis. Antibody microinjection of Aurora-A into late G2 cells induced dose-dependent failure in separation of centriole pairs at prophase, indicating that Aurora-A is essential for proper separation of centriole pairs. When we injected anti-Aurora-A antibodies into prometaphase cells that had separated their centriole pairs, chromosomes were severely misaligned on the metaphase plate, indicating that Aurora-A is required for proper movement of chromosomes on the metaphase plate. Furthermore, inhibition of Aurora-A at metaphase by microinjected antibodies prevented cells from completing cytokinesis, suggesting that Aurora-A also has important functions in late mitosis. These results strongly suggest that Aurora-A is essential for many crucial events during mitosis and that the phosphorylation of a series of substrates by Aurora-A at different stages of mitosis may promote diverse critical events in mitosis to maintain chromosome integrity in human cells. PMID- 14523001 TI - Transcriptional control of the arginine/lysine transporter, cat-1, by physiological stress. AB - Cells respond to physiological stress by phosphorylating the alpha subunit of the translation initiation factor eIF2. This adaptive response inhibits protein synthesis and up-regulates genes essential for cell survival. Cat-1, the transporter for the essential amino acids, arginine and lysine, is one of the up regulated genes. We previously showed that stress increases cat-1 expression by coordinated stabilization of the mRNA and increased mRNA translation. This induction is triggered by amino acid depletion and the unfolded protein response (UPR), which is caused by unfolded proteins in the endoplasmic reticulum. We show here that cat-1 gene transcription is also increased by cellular stress. Our studies demonstrate that the cat-1 gene promoter/regulatory region is TATA-less and is located in a region that includes 94 bases of the first exon. Transcription from this promoter is stimulated 8-fold by cellular stress. An amino acid response element within the first exon is shown to be required for the response to amino acid depletion but not to the UPR. The stimulation of transcription by amino acid depletion requires activation of GCN2 kinase, which phosphorylates eIF2alpha. This phosphorylation also induces translation of the cat-1 mRNA, demonstrating that stress-induced transcriptional and translational control of cat-1 are downstream targets of a signaling pathway initiating with eIF2alpha phosphorylation. Our studies show that the increase in cat-1 gene expression by cellular stress involves at least three types of coordinate regulation: regulation of transcription, regulation of mRNA stability, and regulation of mRNA translation. PMID- 14523002 TI - Glycogen synthase kinase-3beta (GSK3beta) binds to and promotes the actions of p53. AB - The recent discovery of direct interactions between two important regulators of cell fate, the tumor suppressor p53 and glycogen synthase kinase-3beta (GSK3beta), led us to examine the mechanism and outcomes of this interaction. Two regions of p53 were identified that regulate its binding to GSK3beta. Deletion of the p53 activation domain-1 (AD1), but not mutations that prevent MDM2 binding through the AD1 domain, enhanced GSK3beta binding to p53, indicating that the AD1 domain interferes with p53 binding to GSK3beta. Deletion of the p53 basic domain (BD) abrogated GSK3beta binding, and a ten amino acid region within the C terminal BD domain was identified as necessary for binding to GSK3beta. GSK3beta activity was not required for p53 binding, but inhibition of GSK3beta stabilized the association, suggesting a transient interaction during which active GSK3beta promotes actions of p53. This regulatory role of GSK3beta was demonstrated by large reductions of p53-induced increases in the levels of MDM2, p21, and Bax when GSK3beta was inhibited. Besides promoting p53-mediated transcription, GSK3beta also contributed to mitochondrial p53 apoptotic signaling. After DNA damage, mitochondrial GSK3beta co-immunoprecipitated with p53 and was activated, and inhibition of GSK3beta blocked cytochrome c release and caspase-3 activation. Thus, GSK3beta interacts with p53 in both the nucleus and mitochondria and promotes its actions at both sites. PMID- 14523003 TI - ATP potentiates interleukin-1 beta-induced MMP-9 expression in mesangial cells via recruitment of the ELAV protein HuR. AB - Renal mesangial cells express high levels of matrix metalloproteinase 9 (MMP-9) in response to inflammatory cytokines such as interleukin (IL)-1 beta. We demonstrate here that the stable ATP analog adenosine 5'-O-(thiotriphosphate) (ATP gamma S) potently amplifies the cytokine-induced gelatinolytic content of mesangial cells mainly by an increase in the MMP-9 steady-state mRNA level. A Luciferase reporter gene containing 1.3 kb of the MMP-9 5'-promoter region showed weak responses to ATP gamma S but conferred a strong ATP-dependent increase in Luciferase activity when under the additional control of the 3'-untranslated region of MMP-9. By in vitro degradation assay and actinomycin D experiments we found that ATP gamma S potently delayed the decay of MMP-9 mRNA. Gel-shift and supershift assays demonstrated that three AU-rich elements (AREs) present in the 3'-untranslated region of MMP-9 are constitutively bound by complexes containing the mRNA stabilizing factor HuR. The RNA binding of these complexes was markedly increased by ATP gamma S. Mutation of each ARE element strongly impaired the RNA binding of the HuR containing complexes. Reporter gene assays revealed that mutation of one ARE did not affect the stimulatory effects by ATP gamma S, but mutation of all three ARE motifs caused a loss of ATP-dependent increase in luciferase activity without affecting IL-1 beta-inducibility. By confocal microscopy we demonstrate that ATP gamma S increased the nucleo cytoplasmic shuttling of HuR and caused an increase in the cytosolic HuR level as shown by cell fractionation experiments. Together, our results indicate that the amplification of MMP-9 expression by extracellular ATP is triggered through mechanisms that likely involve a HuR-dependent rise in MMP-9 mRNA stability. PMID- 14523004 TI - pH-induced conformational changes of AcrA, the membrane fusion protein of Escherichia coli multidrug efflux system. AB - The multidrug efflux system AcrA-AcrB-TolC of Escherichia coli expels a wide range of drugs directly into the external medium from the bacterial cell. The mechanism of the efflux process is not fully understood. Of an elongated shape, AcrA is thought to span the periplasmic space coordinating the concerted operation of the inner and outer membrane proteins AcrB and TolC. In this study, we used site-directed spin labeling (SDSL) EPR (electron paramagnetic resonance) spectroscopy to investigate the molecular conformations of AcrA in solution. Ten AcrA mutants, each with an alanine to cysteine substitution, were engineered, purified, and labeled with a nitroxide spin label. EPR analysis of spin-labeled AcrA variants indicates that the side chain mobilities are consistent with the predicted secondary structure of AcrA. We further demonstrated that acidic pH induces oligomerization and conformational change of AcrA, and that the structural changes are reversible. These results suggest that the mechanism of action of AcrA in drug efflux is similar to the viral membrane fusion proteins, and that AcrA actively mediates the efflux of substrates. PMID- 14523005 TI - Pse1p mediates the nuclear import of the iron-responsive transcription factor Aft1p in Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae, the iron-responsive transcription factor Aft1p plays a critical role in maintaining iron homeostasis. The activity of Aft1p is induced in response to iron starvation and as a consequence the expression of the iron regulon is increased. We have shown previously that Aft1p is localized to the cytoplasm under iron-replete conditions but that it is localized to the nucleus under iron-depleted conditions. In this study, we identified the transport receptor that mediates the import of Aft1p into the nucleus, located the nuclear localization signal (NLS) sequences of Aft1p, and examined whether the nuclear import of Aft1p is affected by iron status. In pse1-1 cells, which bear a temperature-sensitive mutation of PSE1, Aft1p was misdirected to the cytoplasm during iron starvation at the restrictive temperature. Aft1p could also directly bind to Pse1p and was dissociated from the complex by Ran-GTP in vitro. These results indicate that Aft1p is imported into the nucleus by Pse1p. Supporting this is that the induction of an Aft1p target gene, FTR1, in response to iron starvation was greatly reduced in pse1-1 cells. Furthermore, we demonstrated that the nuclear localization of a mutant Aft1 protein that contains an NLS derived from SV40 was regulated by iron status regardless of whether Pse1p could interact with Aft1p. This suggests that the interaction between Aft1p and Pse1p is not a critical step that controls the iron-regulated nucleo-cytoplasmic transport of Aft1p. PMID- 14523006 TI - Shedding of membrane vesicles mediates fibroblast growth factor-2 release from cells. AB - Fibroblast growth factor-2 (FGF-2), a polypeptide with regulatory activity on cell growth and differentiation, lacks a conventional secretory signal sequence, and its mechanism of release from cells remains unclear. We characterized the role of extracellular vesicle shedding in FGF-2 release. Viable cells released membrane vesicles in the presence of serum. However, in serum-free medium vesicle shedding was dramatically down-regulated, and the cells did not release FGF-2 activity into their conditioned medium. Addition of serum to serum-starved cells rapidly induced intracellular FGF-2 clustering under the plasma membrane and into granules that colocalized with patches of the cell membrane with typical features of shed vesicle membranes. Shed vesicles carried three FGF-2 isoforms (18, 22, 24 kDa). Addition of vesicles to endothelial cells stimulated chemotaxis and urokinase plasminogen activator production, which were blocked by anti-FGF-2 antibodies. Treatment of intact vesicles with 2.0 m NaCl or heparinase, which release FGF-2 from membrane-bound proteoglycans, did not abolish their stimulatory effect on endothelial cells, indicating that FGF-2 is carried inside vesicles. The comparison of the stimulatory effects of shed vesicles and vesicle free conditioned medium showed that vesicles represent a major reservoir of FGF 2. Thus, FGF-2 can be released from cells through vesicle shedding. PMID- 14523007 TI - Oxidized phospholipids induce expression of human heme oxygenase-1 involving activation of cAMP-responsive element-binding protein. AB - Heme oxygenase-1 (HO-1) catalyzes the rate-limiting step in heme degradation, protects against oxidative stress, and shows potent anti-inflammatory effects. Oxidized phospholipids, which are generated during inflammation and apoptosis, modulate the inflammatory response by inducing the expression of several genes including HO-1. Here we investigated the signaling pathways and transcriptional events involved in the induction of HO-1 gene expression by oxidized 1-palmitoyl 2-arachidonoyl-sn-glycero-3-phosphorylcholine (OxPAPC) in human umbilical vein endothelial cells. OxPAPC up-regulated HO-1 mRNA and protein in a time- and concentration-dependent manner, whereas pro-inflammatory agents like TNF-alpha and lipopolysaccharide did not significantly induce HO-1 expression in human umbilical vein endothelial cells. Signaling pathways involved in the OxPAPC mediated HO-1 induction included protein kinases A and C, as well as the mitogen activated protein kinases p38 and ERK. The cAMP-responsive element-binding protein (CREB) was phosphorylated via these pathways in response to OxPAPC treatment and expression of a dominant-negative mutant of CREB inhibited OxPAPC induced activity of a human heme oxygenase-1 promoter-driven luciferase reporter construct. We identified a cAMP-responsive element and a Maf recognition element to be involved in the transcriptional activation of the HO-1 promoter by OxPAPC. In gel shift assays we observed binding of CREB to the cAMP-responsive element after OxPAPC treatment. Induction of HO-1 expression by lipid oxidation products via CREB may represent a feedback mechanism to limit inflammation and associated tissue damage. PMID- 14523009 TI - Regulation of necrosis of H9c2 myogenic cells upon transient energy deprivation. Rapid deenergization of mitochondria precedes necrosis and is controlled by reactive oxygen species, stress kinase JNK, HSP72 and ARC. AB - Subjecting myogenic H9c2 cells to transient energy deprivation leads to a caspase independent death with typical features of necrosis. Here we show that the rupture of cytoplasmic membrane, the terminal event in necrosis, is shortly preceded by rapid depolarization of mitochondrial membranes. The rapid deenergization of mitochondria critically depended upon prior generation of reactive oxygen species (ROS) during ATP depletion stage. Accordingly, expression of catalase prevented mitochondrial depolarization and averted subsequent necrosis. Interestingly, trifluoperazine, a compound that protects cells from ischemic insults, prevented necrosis of H9c2 cells through inhibition of ROS production. Other factors that regulated the mitochondrial membrane depolarization and subsequent loss of plasma membrane integrity include a stress kinase JNK activated at early steps of recovery from ATP depletion, as well as an apoptotic inhibitory protein ARC. Accordingly, inhibition of JNK or overexpression of ARC prevented mitochondrial depolarization and rescued H9c2 cells from necrosis. ROS and JNK affected mitochondrial deenergization and necrosis independently of each other since inhibition of ROS production did not prevent activation of JNK, whereas inhibition of JNK did not suppress ROS accumulation. Therefore, JNK activation and ROS production represent two independent pathways that control mitochondrial depolarization and subsequent necrosis of cells subjected to transient energy deprivation. Overexpression of ARC, although preventing mitochondrial depolarization, did not affect either JNK activation or production of ROS. The major heat shock protein Hsp72 inhibited JNK related steps of necrotic pathway but did not affect ROS accumulation. Interestingly, mitochondrial depolarization and subsequent necrosis can be suppressed by an Hsp72 mutant Hsp72DeltaEEVD, which lacks chaperone function but can efficiently suppress JNK activation. Thus, Hsp72 is directly implicated in a signaling pathway, which leads to necrotic death. PMID- 14523008 TI - Translational thermotolerance provided by small heat shock proteins is limited to cap-dependent initiation and inhibited by 2-aminopurine. AB - Heat shock results in inhibition of general protein synthesis. In thermotolerant cells, protein synthesis is still rapidly inhibited by heat stress, but protein synthesis recovers faster than in naive heat-shocked cells, a phenomenon known as translational thermotolerance. Here we investigate the effect of overexpressing a single heat shock protein on cap-dependent and cap-independent initiation of translation during recovery from a heat shock. When overexpressing alphaB crystallin or Hsp27, cap-dependent initiation of translation was protected but no effect was seen on cap-independent initiation of translation. When Hsp70 was overexpressed however, both cap-dependent and -independent translation were protected. This finding indicates a difference in the mechanism of protection mediated by small or large heat shock proteins. Phosphorylation of alphaB crystallin and Hsp27 is known to significantly decrease their chaperone activity; therefore, we tested phosphorylation mutants of these proteins in this system. AlphaB-crystallin needs to be in its non-phosphorylated state to give protection, whereas phosphorylated Hsp27 is more potent in protection than the unphosphorylatable form. This indicates that chaperone activity is not a prerequisite for protection of translation by small heat shock proteins after heat shock. Furthermore, we show that in the presence of 2-aminopurine, an inhibitor of kinases, among which is double-stranded RNA-activated kinase, the protective effect of overexpressing alphaB-crystallin is abolished. The synthesis of the endogenous Hsps induced by the heat shock to test for thermotolerance is also blocked by 2-aminopurine. Most likely the protective effect of alphaB crystallin requires synthesis of the endogenous heat shock proteins. Translational thermotolerance would then be a co-operative effect of different heat shock proteins. PMID- 14523010 TI - Beta-ketoacyl-acyl carrier protein synthase III (FabH) is essential for bacterial fatty acid synthesis. AB - beta-Ketoacyl-acyl carrier protein (ACP) synthase III (KAS III, also called acetoacetyl-ACP synthase) encoded by the fabH gene is thought to catalyze the first elongation reaction (Claisen condensation) of type II fatty acid synthesis in bacteria and plant plastids. However, direct in vivo evidence that KAS III catalyzes an essential reaction is lacking, because no mutant organism deficient in this activity has been isolated. We report the first bacterial strain lacking KAS III, a fabH mutant constructed in the Gram-positive bacterium Lactococcus lactis subspecies lactis IL1403. The mutant strain carries an in-frame deletion of the KAS III active site region and was isolated by gene replacement using a medium supplemented with a source of saturated and unsaturated long-chain fatty acids. The mutant strain is devoid of KAS III activity and fails to grow in the absence of supplementation with exogenous long-chain fatty acids demonstrating that KAS III plays an essential role in cellular metabolism. However, the L. lactis fabH deletion mutant requires only long-chain unsaturated fatty acids for growth, a source of long-chain saturated fatty acids is not required. Because both saturated and unsaturated fatty acids are required for growth when fatty acid synthesis is blocked by biotin starvation (which prevents the synthesis of malonyl-CoA), another pathway for saturated fatty acid synthesis must remain in the fabH deletion strain. Indeed, incorporation of [1-14C]acetate into fatty acids in vivo showed that the fabH mutant retained about 10% of the fatty acid synthetic ability of the wild-type strain and that this residual synthetic capacity was preferentially diverted to the saturated branch of the pathway. Moreover, mass spectrometry showed that the fabH mutant retained low levels of palmitic acid upon fatty acid starvation. Derivatives of the fabH deletion mutant strain were isolated that were octanoic acid auxotrophs consistent with biochemical studies indicating that the major role of FabH is production of short chain fatty acid primers. We also confirmed the essentiality of FabH in Escherichia coli by use of a plasmid-based gene insertion/deletion system. Together these results provide the first genetic evidence demonstrating that FabH conducts the major condensation reaction in the initiation of type II fatty acid biosynthesis in both Gram-positive and Gram-negative bacteria. PMID- 14523011 TI - The platelet-derived growth factor controls c-myc expression through a JNK- and AP-1-dependent signaling pathway. AB - Pro-inflammatory cytokines, environmental stresses, as well as receptor tyrosine kinases regulate the activity of JNK. In turn, JNK phosphorylates Jun members of the AP-1 family of transcription factors, thereby controlling processes as different as cell growth, differentiation, and apoptosis. Still, very few targets of the JNK-Jun pathway have been identified. Here we show that JNK is required for the induction of c-myc expression by PDGF. Furthermore, we identify a phylogenetically conserved AP-1-responsive element in the promoter of the c-myc proto-oncogene that recruits in vivo the c-Jun and JunD AP-1 family members and controls the PDGF-dependent transactivation of the c-myc promoter. These findings suggest the existence of a novel biochemical route linking tyrosine kinase receptors, such as those for PDGF, and c-myc expression through JNK activation of AP-1 transcription factors. They also provide a novel potential mechanism by which both JNK and Jun proteins may exert either their proliferative or apoptotic potential by stimulating the expression of the c-myc proto-oncogene. PMID- 14523012 TI - Identification of the autoantigen HB as the barrier-to-autointegration factor. AB - The HB autoantigen, a 10-kDa DNA-binding protein recognized by autoantibodies only when bound to DNA, was identified by two-dimensional electrophoresis. Silver stained protein spots corresponding to the antigen were excised from two dimensional electrophoresis gels, digested with trypsin, and analyzed by matrix assisted laser desorption/ionization-reflectron time of flight and nano electrospray ionization-ion trap/mass spectrometry. Data base search identified the HB antigen as the barrier-to-autointegration factor, a cellular protein implicated in the cellular cycle that blocks autointegration and promotes intermolecular integration of retrovirus such as the Moloney murine leukemia and the human immunodeficiency type 1 virus. The physicochemical characteristics described for these proteins, their ability to bind double-stranded DNA but not single-stranded DNA, and their nuclear localization confirm that HB and barrier to-autointegration factor are the same protein. PMID- 14523013 TI - The efficiency and specificity of apurinic/apyrimidinic site bypass by human DNA polymerase eta and Sulfolobus solfataricus Dpo4. AB - One of the most common DNA lesions arising in cells is an apurinic/apyrimidinic (AP) site resulting from base loss. Although a template strand AP site impedes DNA synthesis, translesion synthesis (TLS) DNA polymerases can bypass an AP site. Because this bypass is expected to be highly mutagenic because of loss of base coding potential, here we quantify the efficiency and the specificity of AP site bypass by two Y family TLS enzymes, Sulfolobus solfataricus DNA polymerase 4 (Dpo4) and human DNA polymerase eta (Pol eta). During a single cycle of processive DNA synthesis, Dpo4 and Pol eta bypass synthetic AP sites with 13-30 and 10-13%, respectively, of the bypass efficiency for undamaged bases in the same sequence contexts. These efficiencies are higher than for the A family, exonuclease-deficient Klenow fragment of Escherichia coli DNA polymerase I. We then determined AP site bypass specificity for complete bypass, requiring insertion or misalignment at the AP site followed by multiple incorporations using the aberrant primer templates. Although Dpo4, Pol eta, and Klenow polymerase have different fidelity when copying undamaged DNA, bypass of AP sites lacking A or G by all three polymerases is nearly 100% mutagenic. The majority (70-80%) of bypass events made by all three polymerases are insertion of dAMP opposite the AP site. Single base deletion errors comprise 10-25% of bypass events, with other base insertions observed at lower rates. Given that mammalian cells contain five polymerases implicated in TLS, and given that a large number of AP sites are generated per mammalian cell per day, even moderately efficient AP site bypass could be a source of substitution and frameshift mutagenesis in vivo. PMID- 14523014 TI - Two conformations of archaeal Ssh10b. The origin of its temperature-dependent interaction with DNA. AB - The DNA-binding protein Ssh10b from the hyperthermophilic archaeon Sulfolobus shibatae is a member of the Sac10b family, which has been speculated to be involved in the organization of the chromosomal DNA in Archaea. Ssh10b affects the DNA topology in a temperature dependent fashion that has not been reported for any other DNA-binding proteins. Heteronuclear NMR and site-directed mutagenesis were used to analyze the structural basis of the temperature dependent Ssh10b-DNA interaction. The data analysis indicates that two forms of Ssh10b homodimers co-exist in solution, and the slow cis-trans isomerization of the Leu61-Pro62 peptide bond is the key factor responsible for the conformational heterogeneity of the Ssh10b homodimer. The T-form dimer, with the Leu61-Pro62 bond in the trans conformation, dominates at higher temperature, whereas population of the C-form dimer, with the bond in the cis conformation, increases on decreasing the temperature. The two forms of the Ssh10b dimer show the same DNA binding site but have different conformational features that are responsible for the temperature-dependent nature of the Ssh10b-DNA interaction. PMID- 14523015 TI - Modulation of histamine-induced Ca2+ release by protein kinase C. Effects on cytosolic and mitochondrial [Ca2+] peaks. AB - In HeLa cells, histamine induces production of inositol 1,4,5-trisphosphate (InsP3) and release of Ca2+ from the endoplasmic reticulum (ER). Ca2+ release is typically biphasic, with a fast and brief initial phase, followed by a much slower and prolonged one. In the presence of inhibitors of protein kinase C (PKC), including staurosporine and the specific inhibitors GF109203X and Ro-31 8220, the fast phase continued until the ER became fully empty. On the contrary, treatment with phorbol 12,13-dibutyrate inhibited Ca2+ release. Staurosporine had no effect on InsP3-induced Ca2+ release in permeabilized cells and did not modify either histamine-induced InsP3 production. These data suggest that histamine induces Ca2+ release and with a short lag activates PKC to down-regulate it. Consistently, Ca2+ oscillations induced by histamine were increased in amplitude and decreased in frequency in the presence of PKC inhibitors. We show also that mitochondrial [Ca2+] was much more sensitive to changes in ER-Ca2+ release induced by PKC modulation than cytosolic [Ca2+]. PKC inhibitors increased the histamine-induced mitochondrial [Ca2+] peak by 4-fold but increased the cytosolic [Ca2+] peak only by 20%. On the contrary, PKC activation inhibited the mitochondrial [Ca2+] peak by 90% and the cytosolic one by only 50%. Similarly, the combination of PKC inhibitors with the mitochondrial Ca2+ uniporter activator SB202190 led to dramatic increases in mitochondrial [Ca2+] peaks, with little effect on cytosolic ones. This suggests that activation of ER-Ca2+ release by PKC inhibitors could be involved in apoptosis induced by staurosporine. In addition, these mechanisms allow flexible and independent regulation of cytosolic and mitochondrial [Ca2+] during cell stimulation. PMID- 14523016 TI - Smac3, a novel Smac/DIABLO splicing variant, attenuates the stability and apoptosis-inhibiting activity of X-linked inhibitor of apoptosis protein. AB - X-linked inhibitor of apoptosis protein (XIAP), the most potent member of the inhibitor of apoptosis protein (IAP) family, plays a crucial role in the regulation of apoptosis. XIAP is structurally characterized by three baculovirus IAP repeat (BIR) domains that mediate binding to and inhibition of caspases and a RING domain that confers ubiquitin ligase activity. The caspase inhibitory activity of XIAP can be eliminated by the second mitochondria-derived activator of caspases (Smac)/direct IAP-binding protein with low pI (DIABLO) during apoptosis. Here we report the identification and characterization of a novel isoform of Smac/DIABLO named Smac3, which is generated by alternative splicing of exon 4. Smac3 contains an NH2-terminal mitochondrial targeting sequence required for mitochondrial targeting of Smac3 and an IAP-binding motif essential for Smac3 binding to XIAP. Smac3 is released from mitochondria into the cytosol in response to apoptotic stimuli, where it interacts with the second and third BIR domains of XIAP. Smac3 disrupts processed caspase-9 binding to XIAP, promotes caspase-3 activation, and potentiates apoptosis. Strikingly, Smac3, but not Smac/DIABLO, accelerates XIAP auto-ubiquitination and destruction. Smac3-stimulated XIAP ubiquitination is contingent upon the physical association of XIAP with Smac3 and an intact RING domain of XIAP. Smac3-accelerated XIAP destabilization is, at least in part, attributed to its ability to enhance XIAP ubiquitination. Our study demonstrates that Smac3 is functionally additive to, but independent of, Smac/DIABLO. PMID- 14523017 TI - CD38 signaling in T cells is initiated within a subset of membrane rafts containing Lck and the CD3-zeta subunit of the T cell antigen receptor. AB - In this study we present data supporting that most CD38 is pre-assembled in a subset of Brij 98-resistant raft vesicles, which were stable at 37 degrees C, and have relatively high levels of Lck and the CD3-zeta subunit of T cell antigen receptor-CD3 complex in contrast with a Brij 98-soluble pool, where CD38 is associated with CD3-zeta, and Lck is not detected. Our data further indicate that following CD38 engagement, LAT and Lck are tyrosine phosphorylated exclusively in Brij 98-resistant rafts, and some key signaling components translocate into rafts (i.e. Sos and p85-phosphatidylinositol 3-kinase). Moreover, N-Ras results activated within rafts immediately upon CD38 ligation, whereas activated Erk was mainly found in soluble fractions with delayed kinetics respective to Ras activation. Furthermore, full phosphorylation of CD3-zeta and CD3-epsilon only occurs in rafts, whereas partial CD3-zeta tyrosine phosphorylation occurs exclusively in the soluble pool, which correlated with increased levels of c-Cbl tyrosine phosphorylation in the non-raft fractions. Taken together, these results suggest that, unlike the non-raft pool, CD38 in rafts is able to initiate and propagate several activating signaling pathways, possibly by facilitating critical associations within other raft subsets, for example, LAT rafts via its capacity to interact with Lck and CD3-zeta. Overall, these findings provide the first evidence that CD38 operates in two functionally distinct microdomains of the plasma membrane. PMID- 14523018 TI - Interaction of Epstein-Barr virus latent membrane protein 1 with SCFHOS/beta-TrCP E3 ubiquitin ligase regulates extent of NF-kappaB activation. AB - The Epstein-Barr virus latent membrane protein 1 (LMP1) is pivotal in the transforming activity of this virus. We found that the common LMP1-95-8 variant interacts with Homologue of Slimb (HOS), a receptor for the SCFHOS/betaTrCP ubiquitin-protein isopeptide ligase (E3) via one canonical and one cryptic HOS recognition site. These sites are mutated or deleted in the tumor-derived LMP1 Cao variant, which did not bind to HOS. Mutations within these sites on LMP1-95-8 abrogated HOS binding and increased transforming activity of LMP1. HOS did not regulate stability of LMP1-95-8 unless it was mutated to bear additional lysine residues near the cryptic motif. LMP1 proteins that could not bind to HOS exhibited an increased ability to induce IkappaB degradation and NF-kappaB mediated transcription without further increase in activation of IkappaB kinases. Expression of LMP1-95-8 reduced the levels of endogenous HOS available to interact with phosphorylated IkappaBalpha. Degradation of IkappaBalpha and dose dependence of NF-kappaB activation by LMP1-95-8 were promoted by co-expression of HOS. Our data suggest that LMP1-95-8 is a pseudo-substrate of SCFHOS/betaTrCP E3 ubiquitin ligase and that interaction between LMP1 and HOS restricts the extent of LMP1-induced NF-kappaB signaling. We discuss the potential role of this mechanism in transforming and cytostatic effects of LMP1 variants in cells and Epstein-Barr virus-associated tumors. PMID- 14523019 TI - Cofactor binding modulates the conformational stabilities and unfolding patterns of NAD(+)-dependent DNA ligases from Escherichia coli and Thermus scotoductus. AB - DNA ligases are important enzymes required for cellular processes such as DNA replication, recombination, and repair. NAD(+)-dependent DNA ligases are essentially restricted to eubacteria, thus constituting an attractive target in the development of novel antibiotics. Although such a project might involve the systematic testing of a vast number of chemical compounds, it can essentially gain from the preliminary deciphering of the conformational stability and structural perturbations associated with the formation of the catalytically active adenylated enzyme. We have, therefore, investigated the adenylation induced conformational changes in the mesophilic Escherichia coli and thermophilic Thermus scotoductus NAD(+)-DNA ligases, and the resistance of these enzymes to thermal and chemical (guanidine hydrochloride) denaturation. Our results clearly demonstrate that anchoring of the cofactor induces a conformational rearrangement within the active site of both mesophilic and thermophilic enzymes accompanied by their partial compaction. Furthermore, the adenylation of enzymes increases their resistance to thermal and chemical denaturation, establishing a thermodynamic link between cofactor binding and conformational stability enhancement. Finally, guanidine hydrochloride-induced unfolding of NAD(+)-dependent DNA ligases is shown to be a complex process that involves accumulation of at least two equilibrium intermediates, the molten globule and its precursor. PMID- 14523020 TI - High affinity agonistic metal ion binding sites within the melanocortin 4 receptor illustrate conformational change of transmembrane region 3. AB - We created a molecular model of the human melanocortin 4 receptor (MC4R) and introduced a series of His residues into the receptor protein to form metal ion binding sites. We were able to insert micromolar affinity binding sites for zinc between transmembrane region (TM) 2 and TM3 where the metal ion alone was able to activate this peptide binding G-protein-coupled receptor. The exact conformation of the metal ion interactions allowed us to predict the orientation of the helices, and remodeling of the receptor protein indicated that Glu100 and Ile104 in TM2 and Asp122 and Ile125 in TM3 are directed toward a putative area of activation of the receptor. The molecular model suggests that a rotation of TM3 may be important for activation of the MC4R. Previous models of G-protein-coupled receptors have suggested that unlocking of a stabilizing interaction between the DRY motif, in the cytosolic part of TM3, and TM6 is important for the activation process. We suggest that this unlocking process may be facilitated through creation of a new interaction between TM3 and TM2 in the MC4R. PMID- 14523021 TI - Fibronectin protects prostate cancer cells from tumor necrosis factor-alpha induced apoptosis via the AKT/survivin pathway. AB - Integrins are cell surface heterodimeric transmembrane receptors that, in addition to mediating cell adhesion to extracellular matrix proteins modulate cell survival. This mechanism may be exploited in cancer where evasion from apoptosis invariably contributes to cellular transformation. The molecular mechanisms responsible for matrix-induced survival signals begin to be elucidated. Here we report that the inhibitor of apoptosis survivin is expressed in vitro in human prostate cell lines with the highest levels present in aggressive prostate cancer cells such as PC3 and LNCaP-LN3 as well as in vivo in prostatic adenocarcinoma. We also show that interference with survivin in PC3 prostate cancer cells using a Cys84--> Ala dominant negative mutant or survivin antisense cDNA causes nuclear fragmentation, hypodiploidy, cleavage of a 32-kDa proform caspase-3 to active caspase-3, and proteolysis of the caspase substrate poly(ADP-ribose) polymerase. We demonstrate that in the aggressive PC3 cell line, adhesion to fibronectin via beta1 integrins results in up-regulation of survivin and protection from apoptosis induced by tumor necrosis factor-alpha (TNF-alpha). In contrast, survivin is not up-regulated by cell adhesion in the non-tumorigenic LNCaP cell line. Dominant negative survivin counteracts the ability of fibronectin to protect cells from undergoing apoptosis, whereas wild-type survivin protects non-adherent cells from TNF-alpha-induced apoptosis. Evidence is provided that expression of beta1A integrin is necessary to protect non adherent cells transduced with survivin from TNF-alpha-induced apoptosis. In contrast, the beta1C integrin, which contains a variant cytoplasmic domain, is not able to prevent apoptosis induced by TNF-alpha in non-adherent cells transduced with survivin. Finally, we show that regulation of survivin levels by integrins are mediated by protein kinase B/AKT. These findings indicate that survivin is required to maintain a critical anti-apoptotic threshold in prostate cancer cells and identify integrin signaling as a crucial survival pathway against death receptor-mediated apoptosis. PMID- 14523022 TI - Structures of Streptococcus pneumoniae hyaluronate lyase in complex with chondroitin and chondroitin sulfate disaccharides. Insights into specificity and mechanism of action. AB - Streptococcus pneumoniae hyaluronate lyase is a surface enzyme of this Gram positive bacterium. The enzyme degrades hyaluronan and chondroitin/chondroitin sulfates by cleaving the beta1,4-glycosidic linkage between the glycan units of these polymeric substrates. This degradation helps spreading of this bacterial organism throughout the host tissues and facilitates the disease process caused by pneumococci. The mechanism of this degradative process is based on beta elimination, is termed proton acceptance and donation, and involves selected residues of a well defined catalytic site of the enzyme. The degradation of hyaluronan alone is thought to proceed through a processive mode of action. The structures of complexes between the enzyme and chondroitin as well as chondroitin sulfate disaccharides allowed for the first detailed insights into these interactions and the mechanism of action on chondroitins. This degradation of chondroitin/chondroitin sulfates is nonprocessive and is selective for the chondroitin sulfates only with certain sulfation patterns. Chondroitin sulfation at the 4-position on the nonreducing site of the linkage to be cleaved or 2 sulfation prevent degradation due to steric clashes with the enzyme. Evolutionary studies suggest that hyaluronate lyases evolved from chondroitin lyases and still retained chondroitin/chondroitin sulfate degradation abilities while being specialized in the degradation of hyaluronan. The more efficient processive degradation mechanism has come to be preferred for the unsulfated substrate hyaluronan. PMID- 14523023 TI - Proteasomal degradation of Runx2 shortens parathyroid hormone-induced anti apoptotic signaling in osteoblasts. A putative explanation for why intermittent administration is needed for bone anabolism. AB - It is unknown why sustained elevation of parathyroid hormone (PTH) stimulates bone resorption, whereas intermittent administration stimulates bone formation. We show in mice that daily injections of PTH attenuate osteoblast apoptosis, thereby increasing osteoblast number, bone formation rate, and bone mass, but do not affect osteoclast number. In contrast, sustained elevation of PTH, achieved either by infusion or by raising endogenous hormone secretion with a calcium deficient diet, does not affect osteoblast apoptosis but increases osteoclast number. Attenuation of apoptosis by PTH in cultured osteoblastic cells requires protein kinase A-mediated phosphorylation and inactivation of the pro-apoptotic protein Bad as well as transcription of survival genes, like Bcl-2, mediated by CREB (cAMP response element-binding protein) and Runx2. But, PTH also increases proteasomal proteolysis of Runx2. Moreover, the anti-apoptotic effect of PTH is prolonged by inhibition of proteasomal activity, by overexpressing a dominant negative form of the E3 ligase (ubiquitin-protein isopeptide ligase) that targets Runx2 for degradation (Smurf1), or by overexpressing Runx2 itself. The duration of the anti-apoptotic effect of PTH, thus, depends on the level of Runx2, which in turn is decreased by PTH via Smurf1-mediated proteasomal proteolysis. The self limiting nature of PTH-induced survival signaling might explain why intermittent administration of the hormone is required for bone anabolism. PMID- 14523024 TI - GIT1 mediates Src-dependent activation of phospholipase Cgamma by angiotensin II and epidermal growth factor. AB - Critical events for vasoconstrictor and growth factor signal transduction include stimulation of phospholipase Cgamma (PLCgamma) and elevation of intracellular calcium. c-Src has been proposed as a common mediator for these signals activated by both G protein-coupled receptors (GPCRs) and tyrosine kinase-coupled receptors (TKRs). Here we show that the GPCR kinase-interacting protein-1 (GIT1) is a substrate for c-Src that undergoes tyrosine phosphorylation in response to angiotensin II (AngII) and EGF in vascular smooth muscle and 293 cells. GIT1 associates with PLCgamma via the PLCgamma Src homology 2 and 3 domains constitutively, and the interaction is unaltered by AngII and EGF. GIT1 interaction with PLCgamma is required for PLCgamma activation based on inhibition of tyrosine phosphorylation and calcium mobilization after GIT1 knockdown with antisense GIT1 oligonucleotides. GIT1 interacts with PLCgamma via a novel Spa homology domain (SHD) and a coiled-coil domain. Deletion mutation analysis showed that GIT1(SHD) is required for AngII- and EGF-mediated PLCgamma activation (measured by phosphorylation of Tyr783 and inositol 1,4,5-trisphosphate formation). We propose that GIT1 is a novel regulator of PLCgamma function that mediates PLCgamma activation by c-Src and integrates signal transduction by GPCRs and TKRs. PMID- 14523025 TI - Osteocrin, a novel bone-specific secreted protein that modulates the osteoblast phenotype. AB - Although a number of secreted factors have been demonstrated to be bone regulators, none of these are unique to bone. Using a viral-based signal-trap strategy we have identified a novel gene we have termed "osteocrin." A 1280-bp mRNA encodes osteocrin producing a mature protein of 103 amino acids with a molecular mass of 11.4 kDa. Osteocrin shows no homology with any known gene except for two conserved sequence motifs reminiscent of dibasic cleavage sites found in peptide hormone precursors. Immunofluorescence and Western blot analysis confirmed the secretory nature of osteocrin. Two protein species were identified in the medium of cells overexpressing osteocrin, a full-length 11.4 kDa species and a processed approximately 5 kDa species. Mutation of the 76KKKR79 dibasic cleavage site abolished the appearance of this smaller osteocrin fragment. By in situ hybridization in mouse embryos, osteocrin was expressed specifically in Cbfa 1-positive, osteocalcin-negative osteoblasts. Immunohistochemistry on adult mouse bone showed osteocrin localization in osteoblasts and young osteocytes. By Northern blot analysis, osteocrin expression was only detected in bone, expression peaking just after birth and decreasing markedly with age. In primary osteoblastic cell cultures osteocrin expression coincided with matrix formation then decreased in very mature cultures. Treatment of cultures with 1,25 dihydroxyvitamin D3 resulted in a rapid dose-dependent down-regulation of osteocrin expression, suggesting direct regulation. Chronic treatment of primary cultures with osteocrin-conditioned media inhibited mineralization and reduced osteocalcin and alkaline phosphatase expression. These results suggest that osteocrin represents a novel, unique vitamin D-regulated bone-specific protein that appears to act as a soluble osteoblast regulator. PMID- 14523026 TI - Permease recycling and ubiquitination status reveal a particular role for Bro1 in the multivesicular body pathway. AB - Ubiquitination of the yeast Gap1 permease at the plasma membrane triggers its endocytosis followed by targeting to the vacuolar lumen for degradation. We previously identified Bro1 as a protein essential to this down-regulation. In this study, we show that Bro1 is essential neither to ubiquitination nor to the early steps of Gap1 endocytosis. Bro1 rather intervenes at a late step of the multivesicular body (MVB) pathway, after the core components of the endosome associated ESCRT-III protein complex and before or in conjunction with Doa4, the ubiquitin hydrolase mediating protein deubiquitination prior to their incorporation into MVB vesicles. Bro1 markedly differs from other class E vacuolar protein sorting factors involved in MVB sorting as lack of Bro1 leads to recycling of the internalized permease back to the plasma membrane by passing through the Golgi. This recycling seems to be accompanied by deubiquitination of the permease and unexpectedly requires a normal endosome-to-vacuole transport function. PMID- 14523027 TI - Endothelin-1 activates endothelial cell nitric-oxide synthase via heterotrimeric G-protein betagamma subunit signaling to protein jinase B/Akt. AB - Endothelin-1 has dual vasoactive effects, mediating vasoconstriction via ETA receptor activation of vascular smooth muscle cells and vasorelaxation via ETB receptor activation of endothelial cells. Although it is commonly accepted that endothelin-1 binding to endothelial cell ETB receptors stimulates nitric oxide (NO) synthesis and subsequent smooth muscle relaxation, the signaling pathways downstream of ETB receptor activation are unknown. Here, using a model in which we have utilized isolated primary endothelial cells, we demonstrate that ET-1 binding to sinusoidal endothelial cell ETB receptors led to increased protein kinase B/Akt phosphorylation, endothelial cell nitric-oxide synthase (eNOS) phosphorylation, and NO synthesis. Furthermore, eNOS activation was not dependent on tyrosine phosphorylation, and pretreatment of endothelial cells with pertussis toxin as well as overexpression of a dominant negative G-protein-coupled receptor kinase construct that sequesters betagamma subunits inhibited Akt phosphorylation and NO synthesis. Taken together, the data elucidate a G-protein-coupled receptor signaling pathway for ETB receptor-mediated NO production and call attention to the absolute requirement for heterotrimeric G-protein betagamma subunits in this cascade. PMID- 14523029 TI - The growing pains of biodefense. PMID- 14523031 TI - Women's health and clinical trials. AB - Women have traditionally been underrepresented in clinical trials. In order to translate recent advances in our understanding of the molecular and physiological bases of sex differences into new therapeutics and health practices, sound sex specific clinical data are imperative. Since the founding of the Office of Research on Women's Health within the Office of the Director at the NIH in 1990, inequities in federally funded biomedical research, diagnosis, and treatment of diseases affecting women in the US have been reviewed. Discussed herein is the evolution of gender-related research innovations, primarily within the last decade, and strategies and challenges involved in the success of this recent development. PMID- 14523032 TI - Hepatoprotection via the IL-6/Stat3 pathway. AB - Stat3 is a vital transcription factor that is activated downstream of the gp130 receptor, primarily via IL-6 signaling in adult liver. A new study demonstrates that Stat3 provides hepatoprotection against Fas-mediated apoptotic liver damage by two mechanisms: direct inactivation of caspases and reduction of reactive oxygen species. PMID- 14523033 TI - Selectin and selectin ligand binding: a bittersweet attraction. AB - Inhibition of leukocyte migration into target organs has long been an attractive, though challenging, basis for anti-inflammatory strategies. However, to date, the manipulation of leukocyte rolling along blood vessels has not yielded successful new therapies. An important study may now open new avenues in this exciting field of anti-inflammatory therapies by introducing a putative inhibitor of poly-N acetyllactosamine biosynthesis that affects selectin ligand activity and shows efficacy in a rodent skin inflammation model. PMID- 14523034 TI - Finding NEMO: genetic disorders of NF-[kappa]B activation. AB - The pathways between a receptor and transcriptional activation mediated by NF kappaB are complex. The study of human gene mutations that result in dysregulation of these pathways has provided insight into the functions of individual components of the pathway, their interrelations, and the significance of these systems to the organism. PMID- 14523035 TI - Diabetes, microvascular complications, and cardiovascular complications: what is it about glucose? AB - Glycemic control is the primary mediator of diabetic microvascular complications and also contributes to macrovascular complications. A new study (see related article beginning on page 1049) reveals a previously unrecognized association between oxidant activation of poly(ADP ribose) polymerase (PARP) and upregulation of known mediators of glycemic injury. Inhibitors of PARP may have potential therapeutic roles in the prevention of diabetic complications. PMID- 14523036 TI - Stat3 protects against Fas-induced liver injury by redox-dependent and independent mechanisms. AB - Signal transducer and activator of transcription-3 (Stat3) is one of the most important molecules involved in the initiation of liver development and regeneration. In order to investigate the hepatoprotective effects of Stat3, we examined whether Stat3 protects against Fas-mediated liver injury in the mouse. A constitutively activated form of Stat3 (Stat3-C) was adenovirally overexpressed in mouse liver by intravenous injection, and then a nonlethal dose of Fas agonist (Jo2) was injected intraperitoneally into the mouse (0.3 microg/g body wt). Stat3 C dramatically suppressed both apoptosis and necrosis induced by Jo2. In contrast, liver-specific Stat3-knockout mice failed to survive following Jo2 injection. Stat3-C upregulated expression of FLICE inhibitor protein (FLIP), Bcl xL, and Bcl-2, and accordingly downregulated activities of FLICE and caspase-3 that were redox-independent. Interestingly, Stat3-C also upregulated the redox associated protein redox factor-1 (Ref-1) and reduced apoptosis in liver following Jo2 injection by suppressing oxidative stress and redox-sensitive caspase-3 activity. These findings indicate that Stat3 activation protects against Fas-mediated liver injury by inhibiting caspase activities in redox dependent and -independent mechanisms. PMID- 14523037 TI - A novel protective effect of erythropoietin in the infarcted heart. AB - Erythropoietin (EPO) has been shown to protect neurons from ischemic stroke, but can also increase thrombotic events and mortality rates in patients with ischemic heart disease. We reasoned that benefits of EPO might be offset by increases in hematocrit and evaluated the direct effects of EPO in the ischemic heart. We show that preconditioning with EPO protects H9c2 myoblasts in vitro and cardiomyocytes in vivo against ischemic injury. EPO treatment leads to significantly improved cardiac function following myocardial infarction. This protection is associated with mitigation of myocyte apoptosis, translating into more viable myocardium and less ventricular dysfunction. EPO-mediated myocyte survival appears to involve Akt activation. Importantly, cardioprotective effects of EPO were seen without an increase in hematocrit (eliminating oxygen delivery as an etiologic factor in myocyte survival and function), demonstrating that EPO can directly protect the ischemic and infarcted heart. PMID- 14523038 TI - Prevention of leukocyte migration to inflamed skin with a novel fluorosugar modifier of cutaneous lymphocyte-associated antigen. AB - E-selectin and P-selectin on dermal postcapillary venules play critical roles in the migration of effector T cells into inflamed skin. P-selectin glycoprotein ligand-1 (PSGL-1) modified by alpha1,3-fucosyltransferase is the principal selectin ligand on skin-homing T cells and is required for effector T cell entry into inflamed skin. We have previously shown that a fluorinated analog of N acetylglucosamine peracetylated-4-fluorinated-d-glucosamine (4-F-GlcNAc), inhibits selectin ligand expression on human T cell PSGL-1. To analyze 4-F-GlcNAc efficacy in dampening effector T cell migration to inflamed skin, we elicited allergic contact hypersensitivity (CHS) reactions in mice treated with 4-F GlcNAc. We also investigated 4-F-GlcNAc efficacy on lymphocyte E-selectin ligand expression in LNs draining antigen-sensitized skin and on other immunological processes requisite for CHS responses. Our results showed that 4-F-GlcNAc treatment attenuated lymphocyte E-selectin ligand expression in skin-draining LNs and prevented CHS reactions. Significant reductions in inflammatory lymphocytic infiltrate were observed, while pathways related to antigenic processing and presentation and naive T cell recognition within skin-draining LNs were unaffected. These data indicate that 4-F-GlcNAc prevents CHS by inhibiting selectin ligand activity and the capacity of effector T cells to enter antigen challenged skin without affecting the afferent phase of CHS. PMID- 14523039 TI - Congenital sick sinus syndrome caused by recessive mutations in the cardiac sodium channel gene (SCN5A). AB - Sick sinus syndrome (SSS) describes an arrhythmia phenotype attributed to sinus node dysfunction and diagnosed by electrocardiographic demonstration of sinus bradycardia or sinus arrest. Although frequently associated with underlying heart disease and seen most often in the elderly, SSS may occur in the fetus, infant, and child without apparent cause. In this setting, SSS is presumed to be congenital. Based on prior associations with disorders of cardiac rhythm and conduction, we screened the alpha subunit of the cardiac sodium channel (SCN5A) as a candidate gene in ten pediatric patients from seven families who were diagnosed with congenital SSS during the first decade of life. Probands from three kindreds exhibited compound heterozygosity for six distinct SCN5A alleles, including two mutations previously associated with dominant disorders of cardiac excitability. Biophysical characterization of the mutants using heterologously expressed recombinant human heart sodium channels demonstrate loss of function or significant impairments in channel gating (inactivation) that predict reduced myocardial excitability. Our findings reveal a molecular basis for some forms of congenital SSS and define a recessive disorder of a human heart voltage-gated sodium channel. PMID- 14523040 TI - Anti-IL-5 treatment reduces deposition of ECM proteins in the bronchial subepithelial basement membrane of mild atopic asthmatics. AB - Eosinophil-derived TGF-beta has been implicated in remodeling events in asthma. We hypothesized that reduction of bronchial mucosal eosinophils with anti-IL-5 would reduce markers of airway remodeling. Bronchial biopsies were obtained before and after three infusions of a humanized, anti-IL-5 monoclonal antibody (mepolizumab) in 24 atopic asthmatics in a randomized, double-blind, placebo controlled study. The thickness and density of tenascin, lumican, and procollagen III in the reticular basement membrane (RBM) were quantified immunohistochemically by confocal microscopy. Expression of TGF-beta1 mRNA by airway eosinophils was assessed by in situ hybridization, and TGF-beta1 protein was measured in bronchoalveolar lavage (BAL) fluid by ELISA. At baseline, airway eosinophil infiltration and ECM protein deposition was increased in the RBM of asthmatics compared with nonasthmatic controls. Treating asthmatics with anti-IL 5 antibody, which specifically decreased airway eosinophil numbers, significantly reduced the expression of tenascin, lumican, and procollagen III in the bronchial mucosal RBM when compared with placebo. In addition, anti-IL-5 treatment was associated with a significant reduction in the numbers and percentage of airway eosinophils expressing mRNA for TGF-beta1 and the concentration of TGF-beta1 in BAL fluid. Therefore eosinophils may contribute to tissue remodeling processes in asthma by regulating the deposition of ECM proteins. PMID- 14523041 TI - Regulatory functions of CD8+CD28- T cells in an autoimmune disease model. AB - CD8+ T cell depletion renders CD28-deficient mice susceptible to experimental autoimmune encephalomyelitis (EAE). In addition, CD8-/-CD28-/- double-knockout mice are susceptible to EAE. These findings suggest a role for CD8+ T cells in the resistance of CD28-deficient mice to disease. Adoptive transfer of CD8+CD28- T cells into CD8-/- mice results in significant suppression of disease, while CD8+CD28+ T cells demonstrate no similar effect on the clinical course of EAE in the same recipients. In vitro, CD8+CD28- but not CD8+CD28+ T cells suppress IFN gamma production of myelin oligodendrocyte glycoprotein-specific CD4+ T cells. This suppression requires cell-to-cell contact and is dependent on the presence of APCs. APCs cocultured with CD8+CD28- T cells become less efficient in inducing a T cell-dependent immune response. Such interaction prevents upregulation of costimulatory molecules by APCs, hence decreasing the delivery of these signals to CD4+ T cells. These are the first data establishing that regulatory CD8+CD28- T cells occur in normal mice and play a critical role in disease resistance in CD28-/- animals. PMID- 14523042 TI - Inhibition of GAPDH activity by poly(ADP-ribose) polymerase activates three major pathways of hyperglycemic damage in endothelial cells. AB - In this report, we show that hyperglycemia-induced overproduction of superoxide by the mitochondrial electron transport chain activates the three major pathways of hyperglycemic damage found in aortic endothelial cells by inhibiting GAPDH activity. In bovine aortic endothelial cells, GAPDH antisense oligonucleotides activated each of the pathways of hyperglycemic vascular damage in cells cultured in 5 mM glucose to the same extent as that induced by culturing cells in 30 mM glucose. Hyperglycemia-induced GAPDH inhibition was found to be a consequence of poly(ADP-ribosyl)ation of GAPDH by poly(ADP-ribose) polymerase (PARP), which was activated by DNA strand breaks produced by mitochondrial superoxide overproduction. Both the hyperglycemia-induced decrease in activity of GAPDH and its poly(ADP-ribosyl)ation were prevented by overexpression of either uncoupling protein-1 (UCP-1) or manganese superoxide dismutase (MnSOD), which decrease hyperglycemia-induced superoxide. Overexpression of UCP-1 or MnSOD also prevented hyperglycemia-induced DNA strand breaks and activation of PARP. Hyperglycemia induced activation of each of the pathways of vascular damage was abolished by blocking PARP activity with the competitive PARP inhibitors PJ34 or INO-1001. Elevated glucose increased poly(ADP-ribosyl)ation of GAPDH in WT aortae, but not in the aortae from PARP-1-deficient mice. Thus, inhibition of PARP blocks hyperglycemia-induced activation of multiple pathways of vascular damage. PMID- 14523043 TI - IL-10 is critical for Th2 responses in a murine model of allergic dermatitis. AB - We found that mechanical injury to mouse skin, which can be caused by tape stripping, results in rapid induction of IL-10 mRNA. IL-10-/- mice were used to examine the role of IL-10 in a mouse model of allergic dermatitis induced by epicutaneous (EC) sensitization with OVA on tape-stripped skin. Skin infiltration by eosinophils and expression of eotaxin, IL-4, and IL-5 mRNA in OVA-sensitized skin sites were severely diminished in IL-10-/- mice. Following in vitro stimulation with OVA, splenocytes from EC-sensitized IL-10-/- mice secreted significantly less IL-4, but significantly more IFN-gamma, than splenocytes from WT controls. A similar skewing in cytokine secretion profile was observed in the splenocytes of IL-10-/- mice immunized intraperitoneally with OVA. IL-10-/- APCs skewed the in vitro response of OVA T cell receptor (TCR) transgenic T cells towards Th1. Examination of the Th response of WT and IL-10-/- mice immunized with OVA-pulsed WT or IL-10-/- DCs revealed that both DCs and T cells participate in IL-10 skewing of the Th2 response in vivo. These results suggest that IL-10 plays an important role in the Th2 response to antigen and in the development of skin eosinophilia in a murine model of allergic dermatitis. PMID- 14523044 TI - Inhibition of receptor-localized PI3K preserves cardiac beta-adrenergic receptor function and ameliorates pressure overload heart failure. AB - beta-Adrenergic receptor (betaAR) downregulation and desensitization are hallmarks of the failing heart. However, whether abnormalities in betaAR function are mechanistically linked to the cause of heart failure is not known. We hypothesized that downregulation of cardiac betaARs can be prevented through inhibition of PI3K activity within the receptor complex, because PI3K is necessary for betaAR internalization. Here we show that in genetically modified mice, disrupting the recruitment of PI3K to agonist-activated betaARs in vivo prevents receptor downregulation in response to chronic catecholamine administration and ameliorates the development of heart failure with pressure overload. Disruption of PI3K/betaAR colocalization is required to preserve betaAR signaling, since deletion of a single PI3K isoform (PI3Kgamma knockout) is insufficient to prevent the recruitment of other PI3K isoforms and subsequent betaAR downregulation with catecholamine stress. These data demonstrate a specific role for receptor-localized PI3K in the regulation of betaAR turnover and show that abnormalities in betaAR function are associated with the development of heart failure. Thus, a strategy that blocks the membrane translocation of PI3K and leads to the inhibition of betaAR-localized PI3K activity represents a novel therapeutic approach to restore normal betaAR signaling and preserve cardiac function in the pressure overloaded failing heart. PMID- 14523045 TI - Identification of App1 as a regulator of phagocytosis and virulence of Cryptococcus neoformans. AB - Cryptococcus neoformans is a fungal pathogen that, after inhalation, can disseminate to the brain. Host alveolar macrophages (AMs) represent the first defense against the fungus. Once phagocytosed by AMs, fungal cells are killed by a concerted mechanism, involving the host-cellular response. If the cellular response is impaired, phagocytosis of the fungus may be detrimental for the host, since C. neoformans can grow within macrophages. Here, we identified a novel cryptococcal gene encoding antiphagocytic protein 1 (App1). App1 is a cryptococcal cytoplasmic protein that is secreted extracellularly and found in the serum of infected patients. App1 does not affect melanin production, capsule formation, or growth of C. neoformans. Treatment with recombinant App1 inhibited phagocytosis of fungal cells through a complement-mediated mechanism, and Deltaapp1 mutant is readily phagocytosed by AMs. Interestingly, the Deltaapp1 mutant strain showed a decreased virulence in mice deficient for complement C5 (A/Jcr), but it was hypervirulent in mice deficient for T and NK cells (Tgepsilon26). This study identifies App1 as a novel regulator of virulence for C. neoformans, and it highlights that internalization of fungal cells by AMs increases the dissemination of C. neoformans when the host cellular response is impaired. PMID- 14523046 TI - IL-7 enhances peripheral T cell reconstitution after allogeneic hematopoietic stem cell transplantation. AB - We used clinically relevant murine allogeneic bone marrow transplantation (BMT) models to study the mechanisms by which IL-7 administration can improve posttransplant peripheral T cell reconstitution. After transplant we could distinguish two populations of mature donor T cells: (a) alloreactive T cells with decreased expression of CD127 (IL-7 receptor alpha chain) and (b) nonalloreactive T cells, which express CD127 and undergo homeostatic proliferation. IL-7 administration increased the homeostatic proliferation of nonalloreactive T cells, but had no effect on alloreactive T cells and the development of graft-versus-host disease. Allogeneic transplant of purified hematopoietic stem cells and adoptive transfer of thymocytes into lethally irradiated hosts suggested that recent thymic emigrants can undergo homeostatic proliferation and acquire a memory-like phenotype. We found by BrdU pulse-chase, cell cycle, and annexin V analyses that IL-7 administration has significant proliferative and antiapoptotic effects on posttransplant peripheral T cells. We conclude that homeostatic expansion is important for T cell reconstitution after allogeneic BMT and involves both transferred mature T cells and recent thymic emigrants. Apart from its thymopoietic effects, IL-7 promotes peripheral T cell reconstitution through its selective proliferative and antiapoptotic effects on nonalloreactive and de novo-generated T cells, but has no effect on alloreactive T cells. PMID- 14523047 TI - A hypermorphic IkappaBalpha mutation is associated with autosomal dominant anhidrotic ectodermal dysplasia and T cell immunodeficiency. AB - X-linked anhidrotic ectodermal dysplasia with immunodeficiency (XL-EDA-ID) is caused by hypomorphic mutations in the gene encoding NEMO/IKKgamma, the regulatory subunit of the IkappaB kinase (IKK) complex. IKK normally phosphorylates the IkappaB-inhibitors of NF-kappaB at specific serine residues, thereby promoting their ubiquitination and degradation by the proteasome. This allows NF-kappaB complexes to translocate into the nucleus where they activate their target genes. Here, we describe an autosomal-dominant (AD) form of EDA-ID associated with a heterozygous missense mutation at serine 32 of IkappaBalpha. This mutation is gain-of-function, as it enhances the inhibitory capacity of IkappaBalpha by preventing its phosphorylation and degradation, and results in impaired NF-kappaB activation. The developmental, immunologic, and infectious phenotypes associated with hypomorphic NEMO and hypermorphic IKBA mutations largely overlap and include EDA, impaired cellular responses to ligands of TIR (TLR-ligands, IL-1beta, and IL-18), and TNFR (TNF-alpha, LTalpha1/beta2, and CD154) superfamily members and severe bacterial diseases. However, AD-EDA-ID but not XL-EDA-ID is associated with a severe and unique T cell immunodeficiency. Despite a marked blood lymphocytosis, there are no detectable memory T cells in vivo, and naive T cells do not respond to CD3-TCR activation in vitro. Our report highlights both the diversity of genotypes associated with EDA-ID and the diversity of immunologic phenotypes associated with mutations in different components of the NF-kappaB signaling pathway. PMID- 14523048 TI - TGF-beta switches from tumor suppressor to prometastatic factor in a model of breast cancer progression. AB - The TGF-beta signaling network plays a complex role in carcinogenesis because it has the potential to act as either a tumor suppressor or a pro-oncogenic pathway. Currently, it is not known whether TGF-beta can switch from tumor suppressor to pro-oncogenic factor during the course of carcinogenic progression in a single cell lineage with a defined initiating oncogenic event or whether the specific nature of the response is determined by cell type and molecular etiology. To address this question, we have introduced a dominant negative type II TGF-beta receptor into a series of genetically related human breast-derived cell lines representing different stages in the progression process. We show that decreased TGF-beta responsiveness alone cannot initiate tumorigenesis but that it can cooperate with an initiating oncogenic lesion to make a premalignant breast cell tumorigenic and a low-grade tumorigenic cell line histologically and proliferatively more aggressive. In a high-grade tumorigenic cell line, however, reduced TGF-beta responsiveness has no effect on primary tumorigenesis but significantly decreases metastasis. Our results demonstrate a causal role for loss of TGF-beta responsiveness in promoting breast cancer progression up to the stage of advanced, histologically aggressive, but nonmetastatic disease and suggest that at that point TGF-beta switches from tumor suppressor to prometastatic factor. PMID- 14523049 TI - Formula feeding potentiates docosahexaenoic and arachidonic acid biosynthesis in term and preterm baboon neonates. AB - Infant formulas supplemented with docosahexaenoic acid (DHA) and arachidonic acid (ARA) are now available in the United States; however, little is known about the factors that affect biosynthesis. Baboon neonates were assigned to one of four treatments: term, breast-fed; term, formula-fed; preterm (155 of 182 days gestation), formula-fed; and preterm, formula+DHA/ARA-fed. Standard formula had no DHA/ARA; supplemented formula had 0.61%wt DHA (0.3% of calories) and 1.21%wt ARA (0.6% of calories), and baboon breast milk contained 0.68 +/- 0.22%wt DHA and 0.62 +/- 0.12%wt ARA. At 14 days adjusted age, neonates received a combined oral dose of [U-13C]alpha-linolenic acid (LNA*) and [U-13C]linoleic acid (LA*), and tissues were analyzed 14 days after dose. Brain accretion of linolenic acid derived DHA was approximately 3-fold greater for the formula groups than for the breast-fed group, and dietary DHA partially attenuated excess DHA synthesis among preterms. A similar, significant pattern was found in other organs. Brain linoleic acid-derived ARA accretion was significantly greater in the unsupplemented term group but not in the preterm groups compared with the breast fed group. These data show that formula potentiates the biosynthesis/accretion of DHA/ARA in term and preterm neonates compared with breast-fed neonates and that the inclusion of DHA/ARA in preterm formula partially restores DHA/ARA biosynthesis to lower, breast-fed levels. Current formula DHA concentrations are inadequate to normalize long-chain polyunsaturated fatty acids synthesis to that of breast-fed levels. PMID- 14523050 TI - Ceramide-1-phosphate blocks apoptosis through inhibition of acid sphingomyelinase in macrophages. AB - It was reported previously that ceramide-1-phosphate (Cer-1-P) is mitogenic for fibroblasts (Gomez-Munoz, A., P. A. Duffy, A. Martin, L. O'Brien, H-S. Byun, R. Bittman, and D. N. Brindley. 1995. Mol. Pharmacol. 47: 883-889; Gomez-Munoz, A., L. M. Frago, L. Alvarez, and I. Varela-Nieto. 1997. Biochem. J. 325: 435-440). We now show that Cer-1-P prevents cell death in bone-marrow-derived macrophages (BMDMs) after withdrawal of macrophage colony-stimulating factor (M-CSF). Removal of M-CSF is known to induce apoptosis in these cells. Cer-1-P blocked activation of the caspase-9/caspase-3 pathway and prevented DNA fragmentation, indicating that the enhancement of cell survival was due to inhibition of apoptosis. M-CSF deprivation resulted in activation of acid sphingomyelinase (A-SMase), increased ceramide levels, and a decrease in intracellular Cer-1-P. Exogenously added Cer-1 P inhibited A-SMase in intact BMDMs at concentrations that also prevented apoptosis. Cer-1-P also inhibited A-SMase in cell homogenates, suggesting a possible direct physical interaction of Cer-1-P with the enzyme. In conclusion, these data demonstrate that Cer-1-P blocks apoptosis in BMDMs through inhibition of A-SMase, thereby reducing ceramide generation. This adds a new dimension to the understanding of the metabolic interrelationship of ceramides and Cer-1-P, and shows how altering the balance of intracellular levels of these mediators can affect cell survival. PMID- 14523051 TI - Overexpression of APOC1 in obob mice leads to hepatic steatosis and severe hepatic insulin resistance. AB - Obese obob mice with strong overexpression of the human apolipoprotein C1 (APOC1) exhibit excessive free fatty acid (FFA) and triglyceride (TG) levels and severely reduced body weight (due to the absence of subcutaneous adipose tissue) and skin abnormalities. To evaluate the effects of APOC1 overexpression on hepatic and peripheral insulin sensitivity in a less-extreme model, we generated obob mice with mild overexpression of APOC1 (obob/APOC1(+/-)) and performed hyperinsulinemic clamp analysis. Compared with obob littermates, obob/APOC1(+/-) mice showed reduced body weight (-25%) and increased plasma levels of TG (+632%), total cholesterol (+134%), FFA (+65%), glucose (+73%), and insulin (+49%). Hyperinsulinemic clamp analysis revealed severe whole-body and hepatic insulin resistance in obob/APOC1(+/-) mice and, in addition, increased hepatic uptake of FFA and hepatic TG content. Treatment of obob/APOC1(+/-) mice with rosiglitazone strongly improved whole-body insulin sensitivity as well as hepatic insulin sensitivity, despite a further increase of hepatic fatty acid (FA) uptake and a panlobular increase of hepatic TG accumulation. We conclude that overexpression of APOC1 prevents rosiglitazone-induced peripheral FA uptake leading to severe hepatic steatosis. Interestingly, despite rosiglitazone-induced hepatic steatosis, hepatic insulin sensitivity improves dramatically. We hypothesize that the different hepatic fat accumulation and/or decrease in FA intermediates has a major effect on the insulin sensitivity of the liver. PMID- 14523052 TI - PPARalpha controls the intracellular coenzyme A concentration via regulation of PANK1alpha gene expression. AB - Pantothenate kinase (PanK) is thought to catalyze the first rate-limiting step in CoA biosynthesis. The full-length cDNA encoding the human PanK1alpha protein was isolated, and the complete human PANK1 gene structure was determined. Bezafibrate (BF), a hypolipidemic drug and a peroxisome proliferator activator receptor-alpha (PPARalpha) agonist, specifically increased hPANK1alpha mRNA expression in human hepatoblastoma (HepG2) cells as a function of time and dose of the drug, compared with hPANK1beta, hPANK2, and hPANK3, which did not significantly increase. Four putative PPARalpha response elements were identified in the PANKIalpha promoter, and BF stimulated hPANK1alpha promoter activity but did not alter the mRNA half life. Increased hPANK1alpha mRNA resulted in higher hPanK1 protein, localized in the cytoplasm, and elevated PanK enzyme activity. The enhanced hPANK1alpha gene expression translated into increased activity of the CoA biosynthetic pathway and established a higher steady-state CoA level in HepG2 cells. These data are consistent with a key role for PanK1alpha in the control of cellular CoA content and point to the PPARalpha transcription factor as a major factor governing hepatic CoA levels by specific modulation of PANK1alpha gene expression. PMID- 14523053 TI - Effects of atorvastatin versus fenofibrate on apoB-100 and apoA-I kinetics in mixed hyperlipidemia. AB - Kinetics of apo B and apo AI were assessed in 8 patients with mixed hyperlipidemia at baseline and after 8 weeks of atorvastatin 80 mg q.d. and micronised fenofibrate 200 mg q.d. in a cross-over study. Both increased hepatic production and decreased catabolism of VLDL accounted for elevated cholesterol and triglyceride concentrations at baseline. Atorvastatin significantly decreased triglyceride, total, VLDL and LDL cholesterol and apo B concentrations (-65%, 36%, -57%, -40% and -33%, respectively, P<0.05). Kinetic analysis revealed that atorvastatin stimulated the catabolism of apo B containing lipoproteins, enhanced the delipidation of VLDL1 and decreased VLDL1 production. Fenofibrate lowered triglycerides and VLDL cholesterol (-57% and -64%, respectively, P<0.05) due to enhanced delipidation of VLDL1 and VLDL2 and increased VLDL1 catabolism. Changes of HDL particle composition accounted for the increase of HDL cholesterol during atorvastatin and fenofibrate (18% and 23%, P<0.01). Only fenofibrate increased apo AI concentrations through enhanced apo AI synthesis (45%, P<0.05). We conclude that atorvastatin exerts additional beneficial effects on the metabolism of apo B containing lipoproteins unrelated to an increase in LDL receptor activity. Fenofibrate but not atorvastatin increases apo AI production and plasma turnover. PMID- 14523054 TI - Changes in the levels of cerebral and extracerebral sterols in the brain of patients with Alzheimer's disease. AB - 24S-hydroxycholesterol is a side-chain oxidized oxysterol formed in the brain that is continuously crossing the blood-brain barrier to reach the circulation. There may be an opposite flux of 27-hydroxycholesterol, which is formed to a lower extent in the brain than in most other organs. Here we measured cholesterol, lathosterol, 24S- and 27-hydroxycholesterol, and plant sterols in four different brain areas of deceased Alzheimer's disease (AD) patients and controls. 24S-hydroxycholesterol was decreased and 27-hydroxycholesterol increased in all the brain samples from the AD patients. The difference was statistically significant in four of the eight comparisons. The ratio of 27 hydroxycholesterol to 24S-hydroxycholesterol was significantly increased in all brain areas of the AD patients and also in the brains of aged mice expressing the Swedish Alzheimer mutation APP751. Cholesterol 24S-hydroxylase and 27-hydroxylase protein was not significantly different between AD patients and controls. A high correlation was observed between the levels of 24S-hydroxycholesterol and lathosterol in the frontal cortex of the AD patients but not in the controls. Most probably the high levels of 27-hydroxycholesterol are due to increased influx of this steroid over the blood-brain barrier and the lower levels of 24S hydroxycholesterol to decreased production. The high correlation between lathosterol and 24-hydroxycholesterol is consistent with a close coupling between synthesis and metabolism of cholesterol in the frontal cortex of the AD brain. PMID- 14523055 TI - Multicenter phase II trial of weekly paclitaxel for advanced or metastatic breast cancer: the Saitama Breast Cancer Clinical Study Group (SBCCSG-01). AB - OBJECTIVE: Weekly dosing of paclitaxel has been demonstrated to be a well tolerated, feasible and effective administration schedule. In this study, we evaluated the efficacy and safety of weekly paclitaxel in Japanese women with advanced or metastatic breast cancer. METHODS: Seventy-four patients were enrolled in the study. Paclitaxel was administered by 1 h intravenous infusion at a dose of 80 mg/m2 every week. Administration was continued for 3 weeks followed by a 1 week rest. A short premedication, consisting of dexamethasone 10 mg, ranitidine 50 mg and diphenylhydramine 50 mg, was given prior to each dose of paclitaxel. Eligibility criteria included an Eastern Cooperative Oncology Group performance status of 0, 1 or 2 and adequate hematological, hepatic and renal function. RESULTS: Of 74 patients treated and evaluable for toxicities, 70 were evaluable for response. The mean age was 57.7 years. Forty-nine patients (66.2%) had received prior anthracyclines for metastatic diseases. The overall response rate among 74 patients was 40.5%, including 4.1% complete responses and 36.5% partial responses. The median follow-up time was 481 days (range, 24-903 days). The median time to progression was 4.8 months and median overall survival was 15.8 months. The majority of patients tolerated the treatment very well. Although alopecia was observed in most of the patients (93.2%), grade 3 or 4 neutropenia was 10.8% and grade 2 or 3 peripheral neuropathy was 13.5%. CONCLUSION: Weekly paclitaxel as a 1 h infusion was active and generally well tolerated in previously treated patients. Further study of weekly paclitaxel in combination therapy is warranted. PMID- 14523056 TI - Pharmacokinetics of 5-fluorouracil following hepatic intra-arterial infusion in a VX2 hepatic metastasis model. AB - BACKGROUND: Hepatic intra-arterial infusion chemotherapy of 5-fluorouracil (5-FU) or fluorodeoxyuridine (FUDR) has been a treatment option for liver metastasis from colorectal cancer. However, an optimal administration schedule of 5-FU is still controversial. This study was conducted to evaluate a suitable schedule from the viewpoint of 5-FU metabolites and related enzymes. METHODS: 5-FU was infused into the hepatic artery of rabbits having hepatic deposits of VX2 tumor cells in a daily dose of 1, 4, or 8 mg/kg using various schedules. 5-FU, Thymidylate synthase (TS), TS inhibition rate (TSIR), and the amount of fluoro RNA (F-RNA) were measured. RESULTS: A high concentration of 5-FU was detected in the tumors of the group that was administered a dose of 8 mg/kg. TSIR in the tumor was about two-fold higher in the rabbits that were administered a total dose of 8 mg/kg than in those that were administered doses of 4 mg/kg or less. F RNA, ranging from 27 to 36 ng/mg RNA, was detected in the tumor of the rabbits that were administered a total dose of 8 mg/kg. No difference was observed between the short period and the continuous administration schedules of rabbits that were administered a dose of 8 mg/kg of 5-FU. However, DNA synthesis inhibition in normal hepatic tissue was more dependent on the administration schedule than on the total dose of 5-FU because TSIR was significantly higher with shorter periods of drug administration. CONCLUSION: Intermittent bolus administration of large doses of 5-FU might cause more severe hepatic impairment than continuous administration. These results suggest that hepatic intra-arterial infusion of 5-FU should be administered continuously for liver metastasis, although further experiments including a longer administration period of 5-FU are required. PMID- 14523057 TI - A clinical study of PMCJ-9 (Bacillus Calmette-Guerin Connaught strain) treatment of superficial bladder cancer and carcinoma in situ of the bladder. AB - BACKGROUND: Intravesical Bacillus Calmette-Guerin (BCG) is now a standard treatment for Ta, T1 carcinoma and carcinoma in situ (CIS) of the urinary bladder. In Japan, however, only BCG Tokyo 172 strain is commercially available. We therefore designed a clinical study of PMCJ-9 (BCG Connaught strain) for obtaining approval from Japanese Ministry of Health, Labor and Welfare. METHODS: In the phase I-II study, PMCJ-9 40.5, 81 (standard dose overseas) or 121.5 mg in saline was instilled into the bladder of patients with Ta, T1 or CIS once weekly for 8 weeks. The recommended dose was decided and similarly administered in the late phase II study. RESULTS: In the phase I-II study, 49 patients were evaluable for efficacy. The complete response (CR) rates were 60.0% (9/15), 68.2% (15/22) and 75.0% (9/12) in the 40.5, 81 and 121.5 mg groups. The incidence of adverse drug reactions (ADRs) was similar in all groups, but four 121.5 mg group patients developed severe ADRs. Thus, 81 mg was the recommended dose for the late phase II study. In that study, 39 patients were evaluable, showing CR rates of 71.8% (28/39) overall and 61.5% (16/26) and 92.3% (12/13) for the Ta, T1 and CIS cases. The safety was assessed in 42 patients and three (7.1%) were discontinued owing to ADRs. CONCLUSION: The recommended dose for the BCG Connaught strain was decided as 81 mg. PMCJ-9 administration at this dose level weekly for 8 weeks showed a clear antitumor effect and good safety profile against Ta, T1 and CIS transitional cell carcinoma of the bladder. PMID- 14523058 TI - The limited efficacy of methotrexate, actinomycin D and cisplatin (MAP) for patients with advanced testicular cancer. AB - BACKGROUND: Combination chemotherapy of methotrexate, actinomycin D and cisplatin (MAP) is reported to be effective against gestational choriocarcinoma. METHODS: Eight patients with metastatic testicular cancer who had elevated beta-hCG were treated with MAP. They included three refractory cases and two relapsed cases. An additional three patients received MAP as part of the induction therapy. The MAP therapy consisted of methotrexate (10 mg/m2) on days 1-5, actinomycin D (0.01 mg/kg) on days 1-5 and cisplatin (70 mg/m2) on day 1. RESULTS: In all three refractory patients, MAP failed to achieve tumor marker normalization. However, the elevated tumor markers normalized after MAP in the two cases of relapse. Of these two, one patient relapsed again 7 months after MAP and was subsequently salvaged with high-dose chemotherapy. The other patient relapsed and died of the disease 30 months after receiving MAP. Of the three patients who received MAP as part of the induction chemotherapy, one with pure choriocarcinoma achieved tumor marker normalization after MAP and is still alive without disease progression. In the other two patients, MAP failed to achieve marker normalization and the patients received high-dose chemotherapy. The toxicities were mainly bone marrow suppression and mucositis, which were almost acceptable. CONCLUSIONS: The results demonstrated the limited efficacy of MAP as salvage therapy. In addition, the efficacy of MAP as part of induction chemotherapy was negligible. However, there might be some role for MAP as a salvage therapy for patients with pure choriocarcinoma. PMID- 14523059 TI - Morphologic criteria using biopsy specimens to define the risk of gastric cancer in patients with Helicobacter pylori infection. AB - Although Helicobacter pylori (H. pylori) infection is associated with gastric cancer most patients infected with H. pylori do not develop gastric cancer. Gastric biopsy specimens were taken from 49 patients with early intestinal-type gastric cancer and from 49 age- and sex-matched control subjects. Histological features of gastritis were graded according to the updated Sydney System. Mononuclear cell infiltration of the corpus, and intestinal metaplasia of the angle increased the risk of developing gastric cancer. Probability of gastric cancer (PCA) was calculated using these two parameters. The specificities of 70 < PCA for gastric cancer patients, and PCA < 30 for non-cancer subjects were 94 and 95%, respectively. PCA < 20 was observed only in control subjects. Calculation of PCA, which requires only two biopsy specimens, may be useful to define the patients at high or low risk of intestinal-type gastric cancer among patients infected with H. pylori. PMID- 14523060 TI - Four cases of central nervous system involvement of breast malignant lymphoma. AB - Although malignant lymphoma of the breast is a rare disease, we recently experienced a high incidence of central nervous system (CNS) involvement. Thus, we retrospectively reviewed clinical records of 921 patients with breast tumors, treated from 1990 to 2002, to examine CNS involvement. Eight patients were found to have malignant lymphomas during this period. Of these, four patients ranging from 46 to 80 years of age had CNS involvement, one with Burkitt type and three with diffuse large B-cell lymphoma. The patients received surgery and/or chemotherapy and/or radiotherapy. The four other patients without CNS involvement are alive without tumor, whereas three of four patients with CNS involvement died of the disease. It should be noted that CNS involvement in breast lymphoma is not so rare. It is difficult, but important, to determine whether malignant lymphoma of the breast originates in the breast or is of systemic origin. Although brain metastasis could be treated effectively by radiotherapy and/or chemotherapy, the prognosis was poor. Further intensive treatment is required for breast malignant lymphoma with CNS involvement. PMID- 14523061 TI - Ovarian metastasis originating from bronchioloalveolar carcinoma: a rare presentation of lung cancer. AB - Ovarian metastasis originating from bronchioloalveolar carcinoma (BAC) has not been reported previously. We report a 63-year-old Chinese woman who was diagnosed as BAC with pleural metastasis in 1997. Four years later, she complained of vaginal bleeding, and a pelvic mass was discovered by an abdominal computerized tomography scan. Tumor debulking and total hysterectomy with bilateral salpingo oopherectomy were performed. Pathology disclosed well-differentiated adenocarcinoma, with abundant clear cytoplasm, in the ovaries. Furthermore, immunohistochemical staining revealed that the tumor cells from the ovary and pleura were reactive to thyroid transcription factor 1 (TTF-1) and cytokeratin-7 (CK-7) but were negative for cytokeratin-20 (CK-20). The results of immunohistochemical staining, clinical course, and pathological features were compatible with the diagnosis of BAC with ovarian metastasis. In conclusion, to investigate the primary site of a metastatic ovarian cancer, clinicians should not forget the lungs since the incidence of lung cancer in females is increasing. Moreover, a monoclonal antibody panel for TTF-1, CK-7, and CK-20 may facilitate discrimination between primary and metastasized ovarian adenocarcinomas and/or identifying tumors of pulmonary origin. PMID- 14523062 TI - Multiple pulmonary nodules caused by B-cell post-transplant lymphoproliferative disorder after bone marrow transplantation: monitoring Epstein-Barr virus viral load. AB - We report a patient with myelodysplastic syndrome who underwent an allogeneic bone marrow transplantation during the first remission. On day 110 he had a low grade fever and pulmonary nodules, without superficial lymphadenopathy, were observed. The pulmonary nodules gradually increased in size and in number despite administration of anti-fungal and anti-tuberculosis agents. Transbronchial lung biopsy was performed on day 204, yielding a diagnosis of polyclonal post transplant lymphoproliferative disease (PTLD) positive for Epstein-Barr virus (EBV)-encoded RNA (EBER) and CD20. Subsequent measurement of herpesvirus viral load demonstrated a consistent elevation of EBV viral load from day 96 to day 221. After treatment with anti-CD20 monoclonal antibody (rituximab), regression of pulmonary nodules was confirmed and the number of EBV genome copies decreased to a normal range. This case suggests that monitoring the quantitative EBV viral load may be necessary in cases of EBV-associated PTLD, even in cases presenting pulmonary nodules. Solitary pulmonary nodules may be a rare symptom of PTLD, but in such cases, an aggressive approach may be necessary to obtain a correct diagnosis. PMID- 14523063 TI - Complete appendiceal intussusception induced by primary appendiceal adenocarcinoma in tubular adenoma: a case report. AB - A case of complete intussusception induced by appendiceal carcinoma is reported. The patient was a 49-year-old man complaining of rectal bleeding. Barium enema and colonoscopy revealed a cecal polyp; it was interpreted as an inverted appendix with a tumor. Computed tomography showed an invaginated appendix into the cecal cavity. During surgery, the appendix was found to be inverted completely into the cecum; ileocecal resection with regional lymph node dissection was performed. Microscopic examination revealed well-differentiated adenocarcinoma in tubular adenoma. Diagnosis of intussusception with carcinoma of the appendix is often difficult because appendiceal carcinoma with intussusception of the appendix is a rare condition. Although this condition can be diagnosed by radiographic imaging or colonoscopy, computed tomography has also been useful. The clinical manifestation of appendiceal intussusception with primary appendiceal tumor resembles a large cecal polyp, but its treatment differs greatly. Failure to recognize this condition may result in unexpected complications such as consequent peritonitis in case of endoscopic removal. PMID- 14523064 TI - Ionic basis of tonic firing in spinal substantia gelatinosa neurons of rat. AB - Ionic conductances underlying excitability in tonically firing neurons (TFNs) from substantia gelatinosa (SG) were studied by the patch-clamp method in rat spinal cord slices. Ca(2+)-dependent K(+) (K(CA)) conductance sensitive to apamin was found to prolong the interspike intervals and stabilize firing evoked by a sustained membrane depolarization. Suppression of Ca(2+) and K(CA) currents, however, did not abolish the basic pattern of tonic firing, indicating that it was generated by voltage-gated Na(+) and K(+) currents. Na(+) and K(+) channels were further analyzed in somatic nucleated patches. Na(+) channels exhibited fast activation and inactivation kinetics and followed two-exponential time course of recovery from inactivation. The major K(+) current was carried through tetraethylammonium (TEA)-sensitive rapidly activating delayed-rectifier (K(DR)) channels with a slow inactivation. The TEA-insensitive transient A-type K(+) (K(A)) current was very small in patches and was strongly inactivated at resting potential. Block of K(DR) rather than K(A) conductance by 1 mM TEA lowered the frequency and stability of firing. Intracellular staining with biocytin revealed at least three morphological groups of TFNs. Finally, on the basis of present data, we created a model of TFN and showed that Na(+) and K(DR) currents are sufficient to generate a basic pattern of tonic firing. It is concluded that the balanced contribution of all ionic conductances described here is important for generation and modulation of tonic firing in SG neurons. PMID- 14523065 TI - Activity of neurons in cortical area MT during a memory for motion task. AB - We recorded the activity of middle temporal (MT) neurons in 2 monkeys while they compared the directions of motion in 2 sequentially presented random-dot stimuli, sample and test, and reported them as the same or different by pressing one of 2 buttons. We found that MT neurons were active not only in response to the sample and test stimuli but also during the 1,500-ms delay separating them. Most neurons showed a characteristic pattern of activity consisting of a small burst of firing early in the delay, followed by a period of suppression and a subsequent increase in firing rate immediately preceding the presentation of the test stimulus. In a third of the neurons, the activity early in the delay not only reflected the direction of the sample stimulus, but was also related to the range of local directions it contained. During the middle of the delay the majority of neurons were suppressed, consistent with a gating mechanism that could be used to ignore task-irrelevant stimuli. Late in the delay, most neurons showed an increase in response, probably in anticipation of the upcoming test. Throughout most of the delay there was a directional signal in the population of MT neurons, manifested by higher firing rates following the sample moving in the antipreferred direction. Whereas some of these effects may be related to sensory adaptation, others are more likely to represent a more active task-related process. These results support the hypothesis that MT neurons actively participate in the successful execution of all aspects of the task requiring processing and remembering visual motion. PMID- 14523066 TI - Long-lasting activation of rhythmic neuronal activity by a novel mechanosensory system in the crustacean stomatogastric nervous system. AB - Sensory neurons enable neural circuits to generate behaviors appropriate for the current environmental situation. Here, we characterize the actions of a population (about 60) of bilaterally symmetric bipolar neurons identified within the inner wall of the cardiac gutter, a foregut structure in the crab Cancer borealis. These neurons, called the ventral cardiac neurons (VCNs), project their axons through the crab stomatogastric nervous system to influence neural circuits associated with feeding. Brief pressure application to the cardiac gutter transiently modulated the filtering motor pattern (pyloric rhythm) generated by the pyloric circuit within the stomatogastric ganglion (STG). This modulation included an increased speed of the pyloric rhythm and a concomitant decrease in the activity of the lateral pyloric neuron. Furthermore, 2 min of rhythmic pressure application to the cardiac gutter elicited a chewing motor pattern (gastric mill rhythm) generated by the gastric mill circuit in the STG that persisted for < or =30 min. These sensory actions on the pyloric and gastric mill circuits were mimicked by either ventral cardiac nerve or dorsal posterior esophageal nerve stimulation. VCN actions on the STG circuits required the activation of projection neurons in the commissural ganglia. A subset of the VCN actions on these projection neurons appeared to be direct and cholinergic. We propose that the VCN neurons are mechanoreceptors that are activated when food stored in the foregut applies an outward force, leading to the long-lasting activation of projection neurons required to initiate chewing and modify the filtering of chewed food. PMID- 14523067 TI - Reward-predicting activity of dopamine and caudate neurons--a possible mechanism of motivational control of saccadic eye movement. AB - Recent studies have suggested that the basal ganglia are related to motivational control of behavior. To study how motivational signals modulate motor signals in the basal ganglia, we examined activity of midbrain dopamine (DA) neurons and caudate (CD) projection neurons while monkeys were performing a one-direction rewarded version (1DR) of memory-guided saccade task. The cue stimulus indicated the goal position for an upcoming saccade and the presence or absence of reward after the trial. Among four monkeys we studied, three were sensitive to reward such that saccade velocity was significantly higher in the rewarded trials than in the nonrewarded trials; one monkey was insensitive to reward. In the reward sensitive monkeys, both DA and CD neurons responded differentially to reward indicating and no-reward-indicating cues. Thus DA neurons responded with excitation to a reward-indicating cue and with inhibition to a no-reward indicating cue. A group of CD neurons responded to the cue in their response fields (mostly contralateral) and the cue response was usually enhanced when it indicated reward. In the reward-insensitive monkey, DA neurons showed no response to the cue, while the cue responses of CD neurons were not modulated by reward. Many CD neurons in the reward-sensitive monkeys, but not the reward-insensitive monkey, showed precue activity. These results suggest that DA neurons, with their connection to CD neurons, modulate the spatially selective signals in CD neurons in the reward-predicting manner and CD neurons in turn modulate saccade parameters with their polysynaptic connections to the oculomotor brain stem. PMID- 14523068 TI - Role of the tectorial membrane revealed by otoacoustic emissions recorded from wild-type and transgenic Tecta(deltaENT/deltaENT) mice. AB - Distortion product otoacoustic emissions (DPOAE) were recorded from wild-type mice and mutant Tecta(deltaENT/deltaENT) mice with detached tectorial membranes (TM) under combined ketamine/xylaxine anesthesia. In Tecta(deltaENT/deltaENT) mice, DPOAEs could be detected above the noise floor only when the levels of the primary tones exceeded 65 dB SPL. DPOAE amplitude decreased with increasing frequency of the primaries in Tecta(deltaENT/deltaENT) mice. This was attributed to hair cell excitation via viscous coupling to the surrounding fluid and not by interaction with the TM as in the wild-type mice. Local minima and corresponding phase transitions in the DPOAE growth functions occurred at higher DPOAE levels in wild-type than in Tecta(deltaENT/deltaENT) mice. In less-sensitive Tecta(deltaENT/deltaENT) mice, the position of the local minima varied nonsystematically with frequency or no minima were observed. A bell-like dependence of the DPOAE amplitude on the ratio of the primaries was recorded in both wild-type and Tecta(deltaENT/deltaENT) mice. However, the pattern of this dependence was different in the wild-type and Tecta(deltaENT/deltaENT) mice, an indication that the bell-like shape of the DPOAE was produced by a combination of different mechanisms. A nonlinear low-frequency resonance, revealed by nonmonotonicity of the phase behavior, was seen in the wild-type but not in Tecta(deltaENT/deltaENT) mice. PMID- 14523069 TI - Differential cortical and subcortical activations in learning rotations and gains for reaching: a PET study. AB - Previous studies suggest that horizontal reaching movements are planned vectorially with independent specification of direction and extent. The transformation from visual to hand-centered coordinates requires the learning of a task-specific reference frame and scaling factor. We studied learning of a novel reference frame by imposing a screen-cursor rotation and learning of a scaling factor by imposing a novel gain. Previous work demonstrates that rotation and gain learning have different time courses and patterns of generalization. Here we used PET to identify and compare brain areas activated during rotation and gain learning, with a baseline motor-execution task as the subtracted control. Previous work has shown that the time courses of rotation and gain adaptation have a short rapid phase followed by a longer slow phase. We therefore also sought to compare activations associated with the rapid and slower phases of adaptation. We isolated the rapid phase by alternating opposite values of the rotation or gain every 16 movements. The rapid phase of rotation adaptation activated the preSMA. More complete adaptation to the rotation activated right ventral premotor cortex, right posterior parietal cortex, and the left lateral cerebellum. The rapid phase of gain learning only activated subcortical structures: bilateral putamen and left cerebellum. More complete gain learning failed to show any significant activation. We conclude that the time course of rotation adaptation is paralleled by a frontoparietal shift in activated cortical regions. In contrast, early gain adaptation involves only subcortical structures, which we suggest reflects a more automatic process of contextual recalibration of a scaling factor. PMID- 14523070 TI - Hyperexcitability of cultured spinal motoneurons from presymptomatic ALS mice. AB - ALS (amyotrophic lateral sclerosis) is an adult-onset and deadly neurodegenerative disease characterized by a progressive and selective loss of motoneurons. Transgenic mice overexpressing a mutated human gene (G93A) coding for the enzyme SOD1 (Cu/Zn superoxide dismutase) develop a motoneuron disease resembling ALS in humans. In this generally accepted ALS model, we tested the electrophysiological properties of individual embryonic and neonatal spinal motoneurons in culture by measuring a wide range of electrical properties influencing motoneuron excitability during current clamp. There were no differences in the motoneuron resting potential, input conductance, action potential shape, or afterhyperpolarization between G93A and control motoneurons. The relationship between the motoneuron's firing frequency and injected current (f-I relation) was altered. The slope of the f-I relation and the maximal firing rate of the G93A motoneurons were much greater than in the control motoneurons. Differences in spontaneous synaptic input were excluded as a cause of increased excitability. This finding identifies a markedly elevated intrinsic electrical excitability in cultured embryonic and neonatal mutant G93A spinal motoneurons. We conclude that the observed intrinsic motoneuron hyperexcitability is induced by the SOD1 toxic gain-of-function through an aberration in the process of action potential generation. This hyperexcitability may play a crucial role in the pathogenesis of ALS as the motoneurons were cultured from presymptomatic mice. PMID- 14523071 TI - Accumbal neural responses during the initiation and maintenance of intravenous cocaine self-administration. AB - During a chronic extracellular recording session, animals with a history of cocaine self-administration were allowed to initiate drug seeking under drug-free conditions. Later, in the same recording session, animals engaged in intravenous cocaine self-administration. During the drug-free period, 31% of 70 accumbal neurons showed a significant increase in average firing rate in association with either or both the exposure to cues that signaled the onset of cocaine availability and the subsequent onset of drug-seeking behavior. The neurons that showed an average excitatory response during the drug-free period were the only group of neurons that showed an average excitatory phasic response to cocaine reinforced lever presses during the drug self-administration session. A majority of the neurons that were activated during the drug-free period, like the majority of other neurons, showed decreases in average firing in response to self administered cocaine. However, the neurons that were activated during the drug free period maintained a higher rate of firing throughout the self-administration session than did other accumbal neurons. The data of the present study are consistent with the conclusion that accumbal neurons contribute to, or otherwise process, initiation of drug seeking under drug-free conditions and that they do so via primarily excitatory responses. Furthermore, there is continuity between the drug-free and -exposed conditions in neural responses associated with drug seeking. Finally, the data have potential implications for understanding mechanisms that transduce accumbal-mediated drug effects that contribute to drug addiction. PMID- 14523072 TI - Response properties of single neurons in the zebra finch auditory midbrain: response patterns, frequency coding, intensity coding, and spike latencies. AB - The avian mesencephalicus lateralis, dorsalis (MLd) is the auditory midbrain nucleus in which multiple parallel inputs from lower brain stem converge and through which most auditory information passes to reach the forebrain. Auditory processing in the MLd has not been investigated in songbirds. We studied the tuning properties of single MLd neurons in adult male zebra finches. Pure tones were used to examine tonotopy, temporal response patterns, frequency coding, intensity coding, spike latencies, and duration tuning. Most neurons had no spontaneous activity. The tonotopy of MLd is like that of other birds and mammals; characteristic frequencies (CFs) increase in a dorsal to ventral direction. Four major response patterns were found: 1) onset (49% of cells); 2) primary-like (20%); 3) sustained (19%); and 4) primary-like with notch (12%). CFs ranged between 0.9 and 6.1 kHz, matching the zebra finch hearing range and the power spectrum of song. Tuning curves were generally V-shaped, but complex curves, with multiple peaks or noncontiguous excitatory regions, were observed in 22% of cells. Rate-level functions indicated that 51% of nononset cells showed monotonic relationships between spike rate and sound level. Other cells showed low saturation or nonmonotonic responses. Spike latencies ranged from 4 to 40 ms, measured at CF. Spike latencies generally decreased with increasing sound pressure level (SPL), although paradoxical latency shifts were observed in 16% of units. For onset cells, changes in SPL produced smaller latency changes than for cells showing other response types. Results suggest that auditory midbrain neurons may be particularly suited for processing temporally complex signals with a high degree of precision. PMID- 14523073 TI - Differential modulation of CA1 and dentate gyrus interneurons during exploration of novel environments. AB - Parallel recordings of hippocampal principal cells and interneurons were obtained as rats foraged in familiar and adjacent, novel environments. Firing rates of each cell type were assessed as a function of spatial location. Many CA1 interneurons exhibited large decreases in activity in the novel compared with the familiar environment. Dentate gyrus interneurons, however, were much more likely to exhibit large increases in firing in the novel environment. Neither effect was correlated with basic interneuron discharge properties such as degree of theta modulation, baseline firing rate or degree of spatially modulated discharge. Both CA1 and dentate gyrus interneuron rate changes extended into regions of the familiar environment bordering the novel environment. Principal cells in CA1 and dentate gyrus exhibited similar patterns of place specific activity each being indicative of incorporation of novel spatial information into the spatial representation of the familiar environment. The data indicate that inhibitory networks in the CA1 and dentate gyrus areas are modulated in a divergent fashion during the acquisition of novel spatial information and that interneuron activities can be used to detect those regions of an environment subject to redistribution of principal cell spatial activity patterns. PMID- 14523074 TI - Ocular dominance predicts neither strength nor class of disparity selectivity with random-dot stimuli in primate V1. AB - We address two unresolved issues concerning the coding of binocular disparity in primary visual cortex. Experimental studies and theoretical models have suggested a relationship between a cell's ocular dominance, assessed with monocular stimuli, and its tuning to binocular disparity. First, the disparity energy model of disparity selectivity suggests that there should be a correlation between ocular dominance and the strength of disparity tuning. Second, several studies have reported a relationship between ocular dominance and the shape of the disparity tuning curve, with cells dominated by one eye more likely to have disparity tuning of the tuned-inhibitory type. We investigated both of these relationships in single neurons recorded from the primary visual cortex of awake fixating macaques, using dynamic random-dot patterns as a stimulus. To classify disparity tuning curves quantitatively, we develop a new measure of symmetry, which can be applied to any function. We find no evidence for any correlation between ocular dominance and the nature of disparity tuning. This places constraints on the circuitry underlying disparity tuning. PMID- 14523075 TI - Synaptic strength between motoneurons and terminals of the dorsolateral funiculus is regulated by GABA receptors in the turtle spinal cord. AB - The role of GABAA and GABAB receptors in modulation of excitatory synaptic transmission between motoneurons and terminals from dorsolateral funiculus (DLF) was studied in in vitro spinal cord slices of adult turtles. Muscimol--a GABAA receptor agonist--depressed the monosynaptic excitatory postsynaptic potential (EPSP) induced by stimulation of the DLF and shortened its duration. The input resistance and the membrane time constant also were strongly reduced. The input membrane resistance, the amplitude, and the half-width of the EPSP were reduced at the same rate in the presence of muscimol. Bicuculline--a GABAA receptor antagonist--increased the EPSPs amplitude and the input membrane resistance. The EPSP amplitude ratio elicited by a paired-pulse protocol did not change significantly. Our results suggest that muscimol acts mainly by activation of postsynaptic GABAA receptors located on the motoneuron and the synaptic strength on motoneurons may be modulated by tonic activation of postsynaptic GABAA receptors. Baclofen--a GABAB receptor agonist--also depressed DLF-motoneuron synaptic transmission. However, it did not affect the falling phase of the EPSPs or the motoneuron membrane time constant but induced a small decrement in input resistance. In the presence of baclofen, the amplitude ratio produced by a paired pulse protocol increased significantly. This suggests that baclofen decreased the synaptic strength by inhibition of neurotransmitter release from the DLF terminals via activation of presynaptic GABAB receptors. PMID- 14523076 TI - Persistent LIP activity in memory antisaccades: working memory for a sensorimotor transformation. AB - The lateral intraparietal area (LIP) contains neurons that are active during the memory interval of memory saccades. We call these "persistent neurons." Here we study the activity of the persistent neurons in memory antisaccades, "motor" (the saccade is made toward the response field, although the response field is not stimulated visually) and "visual" (the response field is stimulated visually, but the movement is away from the field). Most persistent neurons are active during parts of the memory intervals of both visual and motor memory-antisaccades. Typically, these parts significantly overlap each other and together span the entire memory interval. The amplitude of the activity changes systematically during the memory intervals of visual and motor memory antisaccades. These changes are reflected in an antisaccade differential activity, which turns first to the visual direction and then crosses over to the motor direction. Some persistent neurons appear to show the paradoxical activity previously characterized in visual neurons; paradoxical activity accelerates the transition of the neuron's activity from visual to motor. These observations suggest that the persistent neurons reflect working memory for the computation of the antisaccade sensorimotor transformation. Ensembles of persistent neurons with different response fields may make up modules of working memory. PMID- 14523077 TI - Stiffness of the gerbil basilar membrane: radial and longitudinal variations. AB - Experimental data on the mechanical properties of the tissues of the mammalian cochlea are essential for understanding the frequency- and location-dependent motion patterns that result in response to incoming sound waves. Within the cochlea, sound-induced vibrations are transduced into neural activity by the organ of Corti, the gross motion of which is dependent on the motion of the underlying basilar membrane. In this study we present data on stiffness of the gerbil basilar membrane measured at multiple positions within a cochlear cross section and at multiple locations along the length of the cochlea. A basic analysis of these data using relatively simple models of cochlear mechanics reveals our most important result: the experimentally measured longitudinal stiffness gradient at the middle of the pectinate zone of the basilar membrane (4.43 dB/mm) can account for changes of best frequency along the length of the cochlea. Furthermore, our results indicate qualitative changes of stiffness deflection curves as a function of radial position; in particular, there are differences in the rate of stiffness growth with increasing tissue deflection. Longitudinal coupling within the basilar membrane/organ of Corti complex is determined to have a space constant of 21 microm in the middle turn of the cochlea. The bulk of our data was obtained in the hemicochlea preparation, and we include a comparison of this set of data to data obtained in vivo. PMID- 14523078 TI - Comparison of memory- and visually guided saccades using event-related fMRI. AB - Previous functional imaging studies have shown an increased hemodynamic signal in several cortical areas when subjects perform memory-guided saccades than that when they perform visually guided saccades using blocked trial designs. It is unknown, however, whether this difference results from sensory processes associated with stimulus presentation, from processes occurring during the delay period before saccade generation, or from an increased motor signal for memory guided saccades. We conducted fMRI using an event-related paradigm that separated stimulus-related, delay-related, and saccade-related activity. Subjects initially fixated a central cross, whose color indicated whether the trial was a memory- or a visually guided trial. A peripheral stimulus was then flashed at one of 4 possible locations. On memory-guided trials, subjects had to remember this location for the subsequent saccade, whereas the stimulus was a distractor on visually guided trials. Fixation cross disappearance after a delay period was the signal either to generate a memory-guided saccade or to look at a visual stimulus that was flashed on visually guided trials. We found slightly greater stimulus related activation for visually guided trials in 3 right prefrontal regions and right rostral intraparietal sulcus (IPS). Memory-guided trials evoked greater delay-related activity in right posterior inferior frontal gyrus, right medial frontal eye field, bilateral supplementary eye field, right rostral IPS, and right ventral IPS but not in middle frontal gyrus. Right precentral gyrus and right rostral IPS exhibited greater saccade-related activation on memory-guided trials. We conclude that activation differences revealed by previous blocked experiments have different sources in different areas and that cortical saccade regions exhibit delay-related activation differences. PMID- 14523079 TI - Modularity of motor output evoked by intraspinal microstimulation in cats. AB - We studied the forces produced at the cat's hindpaw by microstimulation of the ipsi- and contralateral lumbar spinal cord in spinal intact alpha-chloralose anesthetized (n = 3) or decerebrate (n = 3) animals. Isometric force and EMG responses were measured at 9-12 limb configurations, with the paw attached to a force transducer and with the hip and femur fixed. The active forces elicited at different limb configurations were summarized as force fields representing the sagittal plane component of the forces produced at the paw throughout the workspace. The forces varied in amplitude over time but the orientations were stable, and the pattern of an active force field was invariant through time. The active force fields divided into four distinct types, and a few of the fields showed convergence to an equilibrium point. The fields were generally produced by coactivation of the hindlimb muscles. In addition, some of the fields were consistent with known spinal reflexes and the stimulation sites producing them were in laminae where the interneurons associated with those reflexes are known to be located. Muscle activation produced by intraspinal stimulation, as assessed by intramuscular EMG activity, was modified with limb configuration, suggesting that the responses were not fixed, but were modified by position-dependent sensory feedback. The force responses may represent basic outputs of the spinal circuitry and may be related to similar spinal primitives found in the frog and rat. PMID- 14523080 TI - Response patterns along an isofrequency contour in cat primary auditory cortex (AI) to stimuli varying in average and interaural levels. AB - The topographical response of a portion of an isofrequency contour in primary cat auditory cortex (AI) to a series of monaural and binaural stimuli was studied. Responses of single neurons to monaural and a matrix of binaural characteristic frequency tones, varying in average binaural level (ABL) and interaural level differences (ILD), were recorded. The topography of responses to monaural and binaural stimuli was appreciably different. Patches of cells that responded monotonically to increments in ABL alternated with patches that responded nonmonotonically to ABL. The patches were between 0.4 and 1 mm in length along an isofrequency contour. Differences were found among monotonic patches and among nonmonotonic patches. Topographically, activated and silent populations of neurons varied with both changes in ILD and changes in ABL, suggesting that the area of responsive units may underlie the coding of sound level and sound location. PMID- 14523081 TI - Information content of auditory cortical responses to time-varying acoustic stimuli. AB - The present study explores the issue of cortical coding by spike count and timing using statistical and information theoretic methods. We have shown in previous studies that neurons in the auditory cortex of awake primates have an abundance of sustained discharges that could represent time-varying signals by temporal discharge patterns or mean firing rates. In particular, we found that a subpopulation of neurons can encode rapidly occurring sounds, such as a click train, with discharges that are not synchronized to individual stimulus events, suggesting a temporal-to-rate transformation. We investigated whether there were stimulus-specific temporal patterns embedded in these seemingly random spike times. Furthermore, we quantitatively analyzed the precision of spike timing at stimulus onset and during ongoing acoustic stimulation. The main findings are the following. 1) Temporal and rate codes may operate at separate stimulus domains or encode the same stimulus domain in parallel via different neuronal populations. 2) Spike timing was crucial to encode stimulus periodicity in "synchronized" neurons. 3) "Nonsynchronized" neurons showed little stimulus-specific spike timing information in their responses to time-varying signals. Such responses therefore represent processed (instead of preserved) information in the auditory cortex. And 4) spike timing on the occurrence of acoustic events was more precise at the first event than at successive ones and more precise with sparsely distributed events (longer time intervals between events) than with densely packed events. These results indicate that auditory cortical neurons mark sparse acoustic events (or onsets) with precise spike timing and transform rapidly occurring acoustic events into firing rate-based representations. PMID- 14523082 TI - Calbindin influences response to photic input in suprachiasmatic nucleus. AB - It is well known that light resets the circadian clock only at specific times of day. The mechanisms mediating such gating of environmental input to the CNS are not well understood. We show that calbindinD28K (CalB)-containing cells of the suprachiasmatic nucleus (SCN), which are directly retinorecipient, gate photic entrainment of cellular circadian oscillators and thereby determine the timing of locomotor rhythmicity. Specifically, we demonstrate a circadian rhythm of subcellular localization of CalB: whereas the protein is detected at all times in the cytoplasm, it is low or absent in the nucleus during the night. Under normal circumstances, light-induced behavioral phase shifts and Period (Per) gene expression in the SCN occur only during the subjective night. Surprisingly, both behavioral phase shifts and light-induced Per are blocked during the subjective night and enhanced during the subjective day after administration of CalB antisense oligodeoxynucleotides. These results suggest a cellular basis for temporal gating of photic input to the circadian clock. PMID- 14523083 TI - Heterogeneous modulation of place cell firing by changes in context. AB - Hippocampal place cells show spatially localized activity that can be modulated by both geometric information (e.g., the distances and directions of features in the environment) and nongeometric information (e.g., colors, odors, and possibly behaviors). Nongeometric information may allow the discrimination of different spatial contexts. Understanding how nongeometric (or contextual) information affects hippocampal activity is important in light of proposals that the hippocampus may play a role in constructing a representation of spatial context. We investigated the contextual modulation of place cell activity by recording hippocampal place cells while rats foraged in compound contexts comprising black or white color paired with lemon or vanilla odor. Some cells responded to the color or odor changes alone, but most responded to varying combinations of both. Thus, we demonstrate, for the first time, that there is a heterogeneous input by contextual inputs into the hippocampus. We propose a model of contextual remapping of place cells in which the geometric inputs are selectively activated by subsets of contextual stimuli. Because it appears that different place cells are affected by different subsets of contextual stimuli, the representation of the entire context would require activity at the population level, supporting a role for the hippocampus in constructing a representation of spatial context. PMID- 14523084 TI - Transcriptome fingerprints distinguish hallucinogenic and nonhallucinogenic 5 hydroxytryptamine 2A receptor agonist effects in mouse somatosensory cortex. AB - Most neuropharmacological agents and many drugs of abuse modulate the activity of heptahelical G-protein-coupled receptors. Although the effects of these ligands result from changes in cellular signaling, their neurobehavioral activity may not correlate with results of in vitro signal transduction assays. 5 Hydroxytryptamine 2A receptor (5-HT2AR) partial agonists that have similar pharmacological profiles differ in the behavioral responses they elicit. In vitro studies suggest that different agonists acting at the same receptor may establish distinct patterns of signal transduction. Testing this hypothesis in the brain requires a global signal transduction assay that is applicable in vivo. To distinguish the cellular effects of the different 5-HT2AR agonists, we developed an assay for global signal transduction on the basis of high throughput quantification of rapidly modulated transcripts. Study of the responses to agonists in human embryonic kidney 293 cells stably expressing 5-HT2ARs demonstrated that each agonist elicits a distinct transcriptome fingerprint. We therefore studied behavioral and cortical signal transduction responses in wild type and 5-HT2AR null-mutant mice. The hallucinogenic chemicals (+/-)-2,5 dimethoxy-4-iodoamphetamine (DOI) and lysergic acid diethylamide (LSD) stimulated a head-twitch behavioral response that was not observed with the nonhallucinogenic lisuride hydrogen maleate (LHM) and was absent in receptor null mutant mice. We also found that DOI, LSD, and LHM each induced distinct transcriptome fingerprints in somatosensory cortex that were absent in 5-HT2AR null-mutants. Moreover, DOI and LSD showed similarities in the transcriptome fingerprints obtained that were not observed with the behaviorally inactive drug LHM. Our results demonstrate that chemicals acting at the 5-HT2AR induce specific cellular response patterns in vivo that are reflected in unique changes in the somatosensory cortex transcriptome. PMID- 14523085 TI - In vivo assessment of brain interstitial fluid with microdialysis reveals plaque associated changes in amyloid-beta metabolism and half-life. AB - Soluble amyloid-beta (Abeta) peptide converts to structures with high beta-sheet content in Alzheimer's disease (AD). Soluble Abeta is released by neurons into the brain interstitial fluid (ISF), in which it can convert into toxic aggregates. Because assessment of ISF Abeta levels may provide unique insights into Abeta metabolism and AD, an in vivo microdialysis technique was developed to measure it. Our Abeta microdialysis technique was validated ex vivo with human CSF and then in vivo in awake, freely moving mice. Using human amyloid precursor protein (APP) transgenic mice, we found that, before the onset of AD-like pathology, ISF Abeta in hippocampus and cortex correlated with levels of APP in those tissues. After the onset of Abeta deposition, significant changes in the ISF Abeta40/Abeta42 ratio developed without changes in Abeta1-x. These changes differed from changes seen in tissue lysates from the same animals. By rapidly inhibiting Abeta production, we found that ISF Abeta half-life was short ( approximately 2 hr) in young mice but was twofold longer in mice with Abeta deposits. This increase in half-life, without an increase in steady-state levels, suggests that inhibition of Abeta synthesis reveals a portion of the insoluble Abeta pool that is in dynamic equilibrium with ISF Abeta. This now measurable in vivo pool is a likely target for new diagnostic and therapeutic strategies. PMID- 14523086 TI - Cardiotrophin-like cytokine/cytokine-like factor 1 is an essential trophic factor for lumbar and facial motoneurons in vivo. AB - The ciliary neurotrophic factor alpha-receptor (CNTFRalpha) is required for motoneuron survival during development, but the relevant ligand(s) has not been determined. One candidate is the heterodimer formed by cardiotrophin-like cytokine (CLC) and cytokine-like factor 1 (CLF). CLC/CLF binds to CNTFRalpha and enhances the survival of developing motoneurons in vitro; whether this novel trophic factor plays a role in neural development in vivo has not been tested. We examined motor and sensory neurons in embryonic chicks treated with CLC and in mice with a targeted deletion of the clf gene. Treatment with CLC increased the number of lumbar spinal cord motoneurons that survived the cell death period in chicks. However, this effect was regionally specific, because brachial and thoracic motoneurons were unaffected. Similarly, newborn clf-/- mice exhibited a significant reduction in lumbar motoneurons, with no change in the brachial or thoracic cord. Clf deletion also affected brainstem motor nuclei in a regionally specific manner; the number of motoneurons in the facial but not hypoglossal nucleus was significantly reduced. Sensory neurons of the dorsal root ganglia were not affected by either CLC treatment or clf gene deletion. Finally, mRNA for both clc and clf was found in skeletal muscle fibers of embryonic mice during the motoneuron cell death period. These findings support the view that CLC/CLF is a target-derived factor required for the survival of specific pools of motoneurons. The in vivo actions of CLC and CLF can account for many of the effects of CNTFRalpha on developing motoneurons. PMID- 14523087 TI - A molecular mechanism for mRNA trafficking in neuronal dendrites. AB - Specific neuronal mRNAs are localized in dendrites, often concentrated in dendritic spines and spine synapses, where they are translated. The molecular mechanism of localization is mostly unknown. Here we have explored the roles of A2 response element (A2RE), a cis-acting signal for oligodendrocyte RNA trafficking, and its cognate trans-acting factor, heterogeneous nuclear ribonucleoprotein (hnRNP) A2, in neurons. Fluorescently labeled chimeric RNAs containing A2RE were microinjected into hippocampal neurons, and RNA transport followed using confocal laser scanning microscopy. These RNA molecules, but not RNA lacking the A2RE sequence, were transported in granules to the distal neurites. hnRNP A2 protein was implicated as the cognate trans-acting factor: it was colocalized with RNA in cytoplasmic granules, and RNA trafficking in neurites was compromised by A2RE mutations that abrogate hnRNP A2 binding. Coinjection of antibodies to hnRNP A2 halved the number of trafficking cells, and treatment of neurons with antisense oligonucleotides also disrupted A2RE-RNA transport. Colchicine inhibited trafficking, whereas cells treated with cytochalasin were unaffected, implicating involvement of microtubules rather than microfilaments. A2RE-like sequences are found in a subset of dendritically localized mRNAs, which, together with these results, suggests that a molecular mechanism based on this cis-acting sequence may contribute to dendritic RNA localization. PMID- 14523088 TI - The tissue plasminogen activator-plasminogen proteolytic cascade accelerates amyloid-beta (Abeta) degradation and inhibits Abeta-induced neurodegeneration. AB - Accumulation of the amyloid-beta (Abeta) peptide depends on both its generation and clearance. To better define clearance pathways, we have evaluated the role of the tissue plasminogen activator (tPA)-plasmin system in Abeta degradation in vivo. In two different mouse models of Alzheimer's disease, chronically elevated Abeta peptide in the brain correlates with the upregulation of plasminogen activator inhibitor-1 (PAI-1) and inhibition of the tPA-plasmin system. In addition, Abeta injected into the hippocampus of mice lacking either tPA or plasminogen persists, inducing PAI-1 expression and causing activation of microglial cells and neuronal damage. Conversely, Abeta injected into wild-type mice is rapidly cleared and does not cause neuronal degeneration. Thus, the tPA plasmin proteolytic cascade aids in the clearance of Abeta, and reduced activity of this system may contribute to the progression of Alzheimer's disease. PMID- 14523089 TI - Proteasome inhibition stabilizes tau inclusions in oligodendroglial cells that occur after treatment with okadaic acid. AB - Tau-positive inclusions in oligodendrocytes are consistent neuropathological features of corticobasal degeneration, progressive supranuclear palsy, and frontotemporal dementias with Parkinsonism linked to chromosome 17. Here we show by immunohistochemistry that tau-positive oligodendroglial inclusion bodies also contain the small heat-shock protein (HSP) alphaB-crystallin but not HSP70. To study the molecular mechanisms underlying inclusion body formation, we engineered an oligodendroglia cell line (OLN-t40) to overexpress the longest human tau isoform. Treatment of OLN-t40 cells with okadaic acid (OA), an inhibitor of protein phosphatase 2A, caused tau hyperphosphorylation and a decrease in the binding of tau to microtubules. Simultaneously, tau-positive aggregates that also stained with the amyloid-binding dye thioflavin-S as well as with antibodies to tau and alphaB-crystallin were detected. However, they were only transiently expressed and were degraded within 24 hr. When the proteasomal apparatus was inhibited by carbobenzoxy-l-leucyl-l-leucyl-l-leucinal (MG-132) after OA treatment, the aggregates were stabilized and were still detectable after 18 hr in the absence of OA. Incubation with MG-132 alone inhibited tau proteolysis and led to the induction of HSPs, including alphaB-crystallin and to its translocation to the perinuclear region, but did not induce the formation of thioflavin-S-positive aggregates. Hence, although tau hyperphosphorylation induced by protein phosphatase inhibition contributes to pathological aggregate formation, only hyperphosporylation of tau followed by proteasome inhibition leads to stable fibrillary deposits of tau similar to those observed in neurodegenerative diseases. PMID- 14523090 TI - Upregulation of sodium channel Nav1.3 and functional involvement in neuronal hyperexcitability associated with central neuropathic pain after spinal cord injury. AB - Spinal cord injury (SCI) can result in hyperexcitability of dorsal horn neurons and central neuropathic pain. We hypothesized that these phenomena are consequences, in part, of dysregulated expression of voltage-gated sodium channels. Because the rapidly repriming TTX-sensitive sodium channel Nav1.3 has been implicated in peripheral neuropathic pain, we investigated its role in central neuropathic pain after SCI. In this study, adult male Sprague Dawley rats underwent T9 spinal contusion injury. Four weeks after injury when extracellular recordings demonstrated hyperexcitability of L3-L5 dorsal horn multireceptive nociceptive neurons, and when pain-related behaviors were evident, quantitative RT-PCR, in situ hybridization, and immunocytochemistry revealed an upregulation of Nav1.3 in dorsal horn nociceptive neurons. Intrathecal administration of antisense oligodeoxynucleotides (ODNs) targeting Nav1.3 resulted in decreased expression of Nav1.3 mRNA and protein, reduced hyperexcitability of multireceptive dorsal horn neurons, and attenuated mechanical allodynia and thermal hyperalgesia after SCI. Expression of Nav1.3 protein and hyperexcitability in dorsal horn neurons as well as pain-related behaviors returned after cessation of antisense delivery. Responses to normally noxious stimuli and motor function were unchanged in SCI animals administered Nav1.3 antisense, and administration of mismatch ODNs had no effect. These results demonstrate for the first time that Nav1.3 is upregulated in second-order dorsal horn sensory neurons after nervous system injury, showing that SCI can trigger changes in sodium channel expression, and suggest a functional link between Nav1.3 expression and neuronal hyperexcitability associated with central neuropathic pain. PMID- 14523091 TI - Dynamic coding of taste stimuli in the brainstem: effects of brief pulses of taste stimuli on subsequent taste responses. AB - Recent studies have suggested that the response profiles of taste-responsive cells in the brainstem may be modulated by inhibitory interactions, potentially originating from activity in peripheral taste nerves. This idea was explored by testing the hypothesis that brief (100 msec) pulses of taste stimuli would alter the responses to subsequently presented tastants in the nucleus of the solitary tract (NTS) of urethane-anesthetized rats. Pulses of taste stimuli, called prepulses, were followed by a 3 sec presentation of the same or different taste stimulus. The prepulse-stimulus interval was either 1 or 5 sec, during which the tongue was rinsed with distilled water. Taste stimuli consisted of 0.1 m NaCl, 0.5 m sucrose, 0.01 m quinine HCl, and 0.01 m HCl. Taste prepulses suppressed (or enhanced) subsequent taste responses in 30 of 49 (61%) units when the prepulse stimulus interval was 1 sec but were ineffective when this interval was 5 sec. Most commonly, NaCl or HCl prepulses attenuated the response to quinine. Control experiments showed that these effects were not attributable to adaptation, mixture effects, or response variability. In 19 (39%) of the units tested, effects of prepulses were large enough to change the order of effectiveness of the taste stimuli. Taste responses in these cells were "dynamically tuned" in that the magnitude of response was a function of the taste stimulus that immediately preceded it. Dynamic tuning may be the result of inhibitory interactions within the NTS; cells that show dynamic tuning may have a unique function in taste coding. PMID- 14523092 TI - Subunit arrangement in P2X receptors. AB - ATP-gated ionotropic receptors (P2X receptors) are distributed widely in the nervous system. For example, a hetero-oligomeric receptor containing both P2X2 and P2X3 subunits is involved in primary afferent sensation. Each subunit has two membrane-spanning domains. We have used disulfide bond formation between engineered cysteines to demonstrate close proximity between the outer ends of the first transmembrane domain of one subunit and the second transmembrane domain of another. After expression in HEK 293 cells of such modified P2X2 or P2X4 subunits, the disulfide bond formation is evident because an ATP-evoked channel opening requires previous reduction with dithiothreitol. In the hetero-oligomeric P2X2/3 receptor the coexpression of doubly substituted subunits with wild-type partners allows us to deduce that the hetero-oligomeric channel contains adjacent P2X3 subunits but does not contain adjacent P2X2 subunits. The results suggest a "head-to-tail" subunit arrangement in the quaternary structure of P2X receptors and show that a trimeric P2X2/3 receptor would have the composition P2X2(P2X3)2. PMID- 14523093 TI - Lobster (Panulirus interruptus) pyloric muscles express the motor patterns of three neural networks, only one of which innervates the muscles. AB - In several systems, including some well studied invertebrate "model" preparations, rapid, rhythmic inputs drive slow muscles. In this situation muscle contractions can summate temporally between motor neuron bursts, tonically contract, and low-pass filter broad-band input. We have investigated how the muscles innervated by each motor neuron type of the rapid, rhythmic (cycle period, approximately 1 sec) lobster pyloric network respond when driven by previously recorded in vitro pyloric network activity from intact stomatogastric nervous systems. Under these conditions the much slower gastric mill and cardiac sac networks of the stomatogastric nervous system are also active and modify pyloric activity. All of the muscles show pyloric timed phasic contractions that ride on a sustained tonic contraction; muscle activity can range from being almost completely phasic to almost completely tonic. The modifications of pyloric neuron activity induced by gastric mill (cycle period, approximately 10 sec) activity result in some pyloric muscles showing prominent, gastric mill-timed, changes in either phasic or tonic contraction amplitude. The strong modification of pyloric neuron activity induced by cardiac sac (cycle period, approximately 60 sec) activity alters the contractions of all pyloric muscles. These changes are sufficient that for some muscles, in some preparations, the primary muscle output is cardiac sac-timed. This is the first work to examine the motor responses of all pyloric muscle classes to spontaneous stomatogastric activity and shows that the pyloric motor pattern is a complex combination of the activities of three neural networks, although only one (the pyloric) innervates the muscles. PMID- 14523094 TI - Organization of the visual cortex in human albinism. AB - In albinism there is an abnormal projection of part of the temporal retina to the visual cortex contralateral to the eye. This projection, together with the normally routed fibers from nasal retina, provides a cortical hemisphere with visual input from more than the normal hemifield of visual space. In many mammalian models of albinism, a possible sensory mismatch in the visual cortex is avoided either by reorganization of the thalamocortical connections to give the abnormal input an exclusive cortical representation, or by the abnormal input being substantially suppressed. In this study we examine, with fMRI, how the human visual cortex topographically maps its input in albinism. We find that the input from temporal retina is not substantially suppressed and forms a retinotopic mapping that is superimposed on the mapping of the nasal retina in striate and extrastriate areas. The abnormal routing of temporal fibers is not total, with the line of decussation shifting to between 6 and 14 degrees into temporal retina. Our results indicate that the abnormal input to visual cortex in human albinism does not undergo topographic reorganization between the thalamus and cortex. Furthermore, the abnormal input is not significantly suppressed in either striate or extrastriate areas. The topographic mapping that we report in human does not conform, therefore, to the commonly observed patterns in other mammals but takes the form of the "true albino" pattern that has been reported rarely in cat and in the only other individual primate studied. PMID- 14523095 TI - Dopaminergic modulation of axon collaterals interconnecting spiny neurons of the rat striatum. AB - Dopamine is a critical modulator of striatal function; its absence produces Parkinson's disease. Most cellular actions of dopamine are still unknown. This work describes the presynaptic actions of dopaminergic receptor agonists on GABAergic transmission between neostriatal projection neurons. Axon collaterals interconnect projection neurons, the main axons of which project to other basal ganglia nuclei. Most if not all of these projecting axons pass through the globus pallidus. Thus, we lesioned the intrinsic neurons of the globus pallidus and stimulated neostriatal efferent axons antidromically with a bipolar electrode located in this nucleus. This maneuver revealed a bicuculline-sensitive synaptic current while recording in spiny cells. D1 receptor agonists facilitated whereas D2 receptor agonists depressed this synaptic current. In contrast, a bicuculline sensitive synaptic current evoked by field stimulation inside the neostriatum was not consistently modulated, in agreement with previous studies. The data are discussed in light of the most recent experimental and modeling results. The conclusion was that inhibition of spiny cells by axon collaterals of other spiny cells is quantitatively important; however, to be functionally important, this inhibition might be conditioned to the synchronized firing of spiny neurons. Finally, dopamine exerts a potentially important role regulating the extent of lateral inhibition. PMID- 14523096 TI - Glycine is used as a transmitter by decrementing expiratory neurons of the ventrolateral medulla in the rat. AB - The medullary respiratory network involves various types of respiratory neurons. The present study focused on possible inhibitory neurons called decrementing expiratory (E-DEC) neurons and aimed to determine whether their transmitter is glycine or GABA. In Nembutal-anesthetized, neuromuscularly blocked, and artificially ventilated rats we labeled E-DEC neurons with Neurobiotin and processed the tissues for detection of mRNA encoding either glycine transporter 2 (GLYT2) as a marker for glycinergic neurons or glutamic acid decarboxylase isoform 67 (GAD67) as a marker for GABAergic neurons, using in situ hybridization. Of 38 E-DEC neurons that were labeled, cranial motoneurons (n = 14), which were labeled as control, were negative for either GLYT2 mRNA (n = 10) or GAD67 mRNA (n = 4). The other E-DEC neurons (n = 24) were non-motoneurons. Sixteen of them were examined for GLYT2 mRNA, and the majority (11 of 16) was GLYT2 mRNA-positive. The remaining E-DEC neurons (n = 8) were examined for GAD67 mRNA, and all of them were GAD67 mRNA-negative. The GLYT2 mRNA-positive E-DEC neurons were located in the ventrolateral medulla spanning the Botzinger complex (BOT), the rostral ventral respiratory group (VRG), and the caudal VRG. We conclude that not only E-DEC neurons of the BOT but also many E-DEC neurons of the VRG are inhibitory and use glycine as a transmitter. Although the present negative data cannot rule out completely the release of GABA or co-release of glycine and GABA from E-DEC neurons, several lines of evidence suggest that the glycinergic process is primarily responsible for the phasic inhibition of the respiratory network during the expiratory phase. PMID- 14523097 TI - The neurofibromatosis 1 gene product neurofibromin regulates pituitary adenylate cyclase-activating polypeptide-mediated signaling in astrocytes. AB - Individuals with the neurofibromatosis 1 (NF1)-inherited tumor predisposition syndrome develop low-grade astrocytomas. The NF1 tumor suppressor gene product neurofibromin exhibits GTPase-activating activity (GAP) toward RAS, such that loss of neurofibromin expression leads to high levels of activated RAS and increased cell proliferation. Previous work has demonstrated that Nf1 inactivation in astrocytes leads to increased cell proliferation in vitro and in vivo, accompanied by increased RAS pathway activation. Studies on Nf1 mutant Drosophila have suggested that neurofibromin might also regulate cAMP signaling. Because intracellular cAMP levels have profound effects on astrocyte growth control, we sought to determine the contribution of neurofibromin to astrocyte cAMP regulation. In this report, we demonstrate that Nf1 inactivation in astrocytes results in reduced cAMP generation in response to PACAP and attenuated calcium influx and Rap1 activation. Based on the differential effects of forskolin and dibutyryl-cAMP on Nf1-/- astrocytes, neurofibromin likely functions at the level of adenylyl cyclase activation. Last, the reintroduction of a fragment of neurofibromin containing residues sufficient for restoring RAS-GAP function in Nf1-/- cells resulted in only partial restoration of neurofibromin mediated cAMP regulation. These results demonstrate that neurofibromin positively influences cAMP generation and activation of cAMP growth regulatory targets in astrocytes and expands the role of the NF1 gene in astrocyte growth regulation. PMID- 14523098 TI - Fine structure and biochemical mechanisms underlying nigrostriatal inclusions and cell death after proteasome inhibition. AB - Mutation of genes encoding for various components of a metabolic pathway named the ubiquitin-proteasome system (UP) leads to inherited forms of Parkinson's disease (PD), whereas various components of the UP are constantly present within neuronal inclusions, Lewy bodies, that characterize most genetic and sporadic forms of PD. It has been hypothesized that impairment of this metabolic pathway might be a common mechanism for the onset of PD, and a recent study demonstrated a dysfunction of the UP system within the substantia nigra of patients affected by sporadic PD. In search for the mechanisms underlying the selective toxicity for nigral neurons after inhibition of the UP system, we explored the selective effects after striatal microinfusions of lactacystin or epoxomycin and potential retrograde changes within the ipsilateral substantia nigra. We found that neurotoxicity was selective for striatal dopamine (DA) components and led to retrograde apoptosis within nigral DA cells, which developed neuronal inclusions staining for antigens of the UP system. We found the same ultrastructural features characterizing inclusions obtained in vivo and in vitro after UP inhibition. In vivo, lactacystin-epoxomycin-induced toxicity was suppressed by inhibiting DA synthesis. Similarly, in vitro inclusions and apoptosis were prevented by reducing endogenous DA. On the other hand, toxicity of proteasome inhibition was enhanced by drugs augmenting DA availability: l-3,4 dihydroxyphenylalanine, monoamine oxidase blockers, and DA beta-hydroxylase blockers. These findings demonstrate that impairment of the UP system produces cell death and neuronal inclusions selectively for DA-containing neurons that depend on the occurrence of endogenous DA. PMID- 14523099 TI - Glutamate and amyloid beta-protein rapidly inhibit fast axonal transport in cultured rat hippocampal neurons by different mechanisms. AB - Impairment of axonal transport leads to neurodegeneration and synapse loss. Glutamate and amyloid beta-protein (Abeta) have critical roles in the pathogenesis of Alzheimer's disease (AD). Here we show that both agents rapidly inhibit fast axonal transport in cultured rat hippocampal neurons. The effect of glutamate (100 microm), but not of Abeta25-35 (20 microm), was reversible, was mimicked by NMDA or AMPA, and was blocked by NMDA and AMPA antagonists and by removal of extracellular Ca2+. The effect of Abeta25-35 was progressive and irreversible, was prevented by the actin-depolymerizing agent latrunculin B, and was mimicked by the actin-polymerizing agent jasplakinolide. Abeta25-35 induced intracellular actin aggregation, which was prevented by latrunculin B. Abeta31-35 but not Abeta15-20 exerted effects similar to those of Abeta25-35. Full-length Abeta1-42 incubated for 7 d, which specifically contained 30-100 kDa molecular weight assemblies, also caused an inhibition of axonal transport associated with intracellular actin aggregation, whereas freshly dissolved Abeta1-40, incubated Abeta1-40, and fresh Abeta1-42 had no effect. These results suggest that glutamate inhibits axonal transport via activation of NMDA and AMPA receptors and Ca2+ influx, whereas Abeta exerts its inhibitory effect via actin polymerization and aggregation. The ability of Abeta to inhibit axonal transport seems to require active amino acid residues, which is probably present in the 31-35 sequence. Full-length Abeta may be effective when it represents a structure in which these active residues can access the cell membrane. Our results may provide insight into the early pathogenetic mechanisms of AD. PMID- 14523102 TI - Staphylococcus research. PMID- 14523100 TI - Inhibitory interconnections control burst pattern and emergent network synchrony in reticular thalamus. AB - Inhibitory connections between neurons of the thalamic reticular (RE) nucleus are thought to help prevent spike-wave discharge (SWD), characteristic of generalized absence epilepsy, in thalamic and thalamocortical circuits. Indeed, oscillations in thalamic slices resemble SWD when intra-RE inhibition is blocked and are suppressed when intra-RE inhibition is enhanced. To elucidate the cellular mechanisms underlying these network changes, we recorded from RE cells during oscillations in thalamic slices and either blocked intra-RE inhibition with picrotoxin or enhanced it with clonazepam. We found that intra-RE inhibition limits the number and synchrony, but not the duration, of RE cell bursts. We then performed simulations that demonstrate how inhibition can shift network activity into a desynchronized mode simply by vetoing occasional RE cell bursts. In contrast, when intra-RE inhibition is blocked, RE cells burst synchronously, enabling even short RE cell bursts to promote epileptogenesis in two ways: first, by activating GABA(B) receptors, and second, through the GABA(B) receptor independent emergence of network synchrony. PMID- 14523101 TI - PS2APP transgenic mice, coexpressing hPS2mut and hAPPswe, show age-related cognitive deficits associated with discrete brain amyloid deposition and inflammation. AB - Transgenic mice, expressing mutant beta-amyloid precursor proteins (betaAPPs), have lead to a better understanding of the pathophysiological processes in Alzheimer's disease (AD). In many of these models, however, the temporal development of cognitive decline and the relationship to Abeta deposition and inflammation are unclear. We now report a novel transgenic mouse line, PS2APP (PS2N141I x APPswe), which develops a severe cerebral amyloidosis in discrete brain regions, and present a cross-sectional analysis of these mice at 4, 8, 12, and 16 months of age. Each age cohort was investigated for changes in behavior, electrophysiology of synapse efficacy, ELISA-determined Abeta load, histopathology, and in immunoelectron microscopy. Cognitive deficits were first observed at 8 months when Abeta deposits and inflammation were restricted to discrete brain regions, namely the subiculum and frontolateral (motor and orbital) cortex. As early as 5 months, electron microscopy revealed the presence, in these regions, of pre-plaque, immunogold-labeled extracellular fibrillar Abeta. At the same age, increased levels of insoluble Abeta were detected by ELISA, with Abeta1-40 levels exceeding those of Abeta1-42. Further cognitive decline occurred in an age-related manner, and this was accompanied by the spread of amyloidosis to ultimately affect not only neo- and limbic cortices, but also thalamic and pontine nuclei. Dentate gyrus post-tetanic potentiation was significantly attenuated at 17 months, and there were also significant differences in paired-pulse parameters. This systematic cross-sectional study of the behavioral and pathological changes in the PS2APP mouse indicates that it develops age-related cognitive decline associated with severe amyloidosis and inflammation in discrete brain regions and therefore is suitable for testing a range of potential symptomatic and disease-modifying therapies for AD. PMID- 14523103 TI - The adhesive and immunomodulating properties of the multifunctional Staphylococcus aureus protein Eap. AB - Adherence of Staphylococcus aureus to the host tissue is an important step in the initiation of pathogenesis. At least 10 adhesins produced by S. aureus have been described and it is becoming clear that the expression of these adhesins and their interactions with eukaryotic cells involve complex processes. Some of these, such as the fibronectin-binding proteins (FnBPs) and Clumping Factor A, are well characterized. However, in the last 10 years a number of novel S. aureus adhesins have been described. Functional analyses of these proteins, one of which is Eap (extracellular adherence protein, also known as Map and p70), are revealing important information on the pathogenesis of staphylococcal disease. More than 10 years after the first report of Eap, we are beginning to understand that this protein, which has a broad spectrum of functions, may be a critical factor in the pathogenesis of S. aureus. This review will focus on the interactions of Eap with eukaryotic cells, plasma proteins and the extracellular matrix as well as on the recently recognized role of Eap as an important mediator in the immune response to staphylococcal infection. PMID- 14523104 TI - The staphylococcal saeRS system coordinates environmental signals with agr quorum sensing. AB - sae is a two-component signal transduction system in Staphylococcus aureus that regulates the expression of many virulence factors at the transcriptional level and appears to act synergistically with agr in some cases. In this study, the interactions between sae and agr have been characterized in some detail. It was found that the sae locus is larger and more complex than originally envisioned, in that it is expressed from several promoters, giving rise to four or five transcripts, at least three of which are initiated upstream of saeRS and contain two additional reading frames, here designated saeP and saeQ, which are likely to have important roles in sae function. The upstream transcripts are induced during exponential phase concomitantly with the onset of RNAIII synthesis and their induction requires the agr effector, RNAIII, but is blocked by several environmental signals that override the effects of RNAIII. saeR is also required for the induction of these transcripts, so that the sae locus contains an autoinduction circuit. It is suggested that sae is downstream of agr in the exoprotein activation pathway (and also epistatic with agr), that it coordinates the effects of environmental signals with the agr quorum-sensing system, and therefore that it is a key intermediary in the overall regulatory strategy by which S. aureus senses and responds to its environment. PMID- 14523105 TI - Genome-wide transcriptional profiling of the response of Staphylococcus aureus to cell-wall-active antibiotics reveals a cell-wall-stress stimulon. AB - The molecular events following inhibition of bacterial peptidoglycan synthesis have not been studied extensively. Previous proteomic studies have revealed that certain proteins are produced in increased amounts upon challenge of Staphylococcus aureus with cell-wall-active antibiotics. In an effort to further those studies, the genes upregulated in their expression in response to cell-wall active antibiotics have been identified by genome-wide transcriptional profiling using custom-made Affymetrix S. aureus GeneChips. A large number of genes, including ones encoding proteins involved in cell-wall metabolism (including pbpB, murZ, fmt and vraS) and stress responses (including msrA, htrA, psrA and hslO), were upregulated by oxacillin, D-cycloserine or bacitracin. This response may represent the transcriptional signature of a cell-wall stimulon induced in response to cell-wall-active agents. The findings imply that treatment with cell wall-active antibiotics results in damage to proteins including oxidative damage. Additional genes in a variety of functional categories were upregulated uniquely by each of the three cell-wall-active antibiotics studied. These changes in gene expression can be viewed as an attempt by the organism to defend itself against the antibacterial activities of the agents. PMID- 14523106 TI - Application of a bacterial two-hybrid system for the analysis of protein-protein interactions between FemABX family proteins. AB - Protein-protein interactions play an important role in all cellular processes. The development of two-hybrid systems in yeast and bacteria allows for in vivo assessment of such interactions. Using a recently developed bacterial two-hybrid system, the interactions of the Staphylococcus aureus proteins FemA, FemB and FmhB, members of the FemABX protein family, which is involved in peptidoglycan biosynthesis and beta-lactam resistance of numerous Gram-positive bacteria, were analysed. While FmhB is involved in the addition of glycine 1 of the pentaglycine interpeptide of S. aureus peptidoglycan, FemA and FemB are specific for glycines 2/3 and 4/5, respectively. FemA-FemA, FemA-FemB and FemB-FemB interactions were found, while FmhB exists solely as a monomer. Interactions detected by the bacterial two-hybrid system were confirmed using the glutathione S-transferase pulldown assay and gel filtration. PMID- 14523107 TI - Multiple methionine sulfoxide reductase genes in Staphylococcus aureus: expression of activity and roles in tolerance of oxidative stress. AB - Staphylococcus aureus contains three genes encoding MsrA-specific methionine sulfoxide reductase (Msr) activity (msrA1, msrA2 and msrA3) and an additional gene that encodes MsrB-specific Msr activity. Data presented here suggest that MsrA1 is the major contributor of the MsrA activity in S. aureus. In mutational analysis, while the total Msr activity in msrA2 mutant was comparable to that of the parent, Msr activity was significantly up-regulated in the msrA1 or msrA1 msrA2 double mutant. Assessment of substrate specificity together with increased reactivity of the cell-free protein extracts of the msrA1 mutants to anti-MsrB polyclonal antibodies in Western analysis provided evidence that increased Msr activity was due to elevated synthesis of MsrB in the MsrA1 mutants. Previously, it was reported that oxacillin treatment of S. aureus cells led to induced synthesis of MsrA1 and a mutation in msrA1 increased the susceptibility of the organism to H(2)O(2). A mutation in the msrA2 gene, however, was not significant for the bacterial oxidative stress response. In complementation assays, while the msrA2 gene was unable to complement the msrA1 msrA2 double mutant for H(2)O(2) resistance, the same gene restored H(2)O(2) tolerance in the double mutant when placed under the control of the msrA1 promoter. However, msrA1 which was able to complement the oxidative stress response in msrA1 mutants could not restore the tolerance of the msrA1 msrA2 mutants to H(2)O(2) when placed under the control of the msrA2 promoter. Additionally, although the oxacillin minimum inhibitory concentration of the msrA1 mutant was comparable to that of the wild-type parent, in shaking liquid culture, the msrA1 mutant responded more efficiently to sublethal doses of oxacillin. The data suggest complex regulation of Msr proteins and a more significant physiological role for msrA1/msrB in S. aureus. PMID- 14523108 TI - Role and regulation of the superoxide dismutases of Staphylococcus aureus. AB - Staphylococcus aureus has two superoxide dismutases (SODs), encoded by the sodA and sodM genes, which inactivate harmful superoxide radicals () encountered during host infection or generated from aerobic metabolism. The transcriptional start sites have been mapped and expression analysis on reporter fusions in both genes has been carried out. Under standard growth conditions, manganese (Mn), a mineral superoxide scavenger, elevated total SOD activity but had no effect on the transcription of either gene. Transcription of sodA and sodM was most strongly induced by either internally or externally generated, respectively. Sensitivity to internally generated was linked with SodA deficiency. Mn supplementation completely rescued a sodA mutant when challenged by internally generated, and this was growth-phase-dependent. Sensitivity to externally generated stress was only observed in a sodA sodM mutant and was Mn-independent. In a mouse abscess model of infection, isogenic sodA, sodM and sodA sodM mutants had reduced virulence compared to the parental strain, showing the importance of the enzymic scavenging system for the survival of the pathogen. PMID- 14523109 TI - The Staphylococcus aureus surface protein SasG and its homologues promote bacterial adherence to human desquamated nasal epithelial cells. AB - Staphylococcus aureus binds to human desquamated nasal epithelial cells, a phenomenon likely to be important in nasal colonization. ClfB was identified previously as one staphylococcal adhesin that promoted binding to nasal epithelia. In this study, it is shown that the S. aureus surface protein SasG, identified previously by in silico analysis of genome sequences, and two homologous proteins, Pls of S. aureus and AAP of Staphylococcus epidermidis, also promote bacterial adherence to nasal epithelial cells. Conditions for in vitro expression of SasG by S. aureus were not found. Adherence assays were therefore performed with S. aureus and Lactococcus lactis expressing SasG from an expression plasmid. These studies showed that SasG did not bind several ligands typically bound by S. aureus. Significantly, SasG and Pls did promote bacterial adherence to nasal epithelial cells. Furthermore, pre-incubation of epithelial cells with purified recombinant proteins revealed that the N-terminal A regions of SasG, Pls and AAP, but not the B repeats of SasG, inhibited adherence of L. lactis expressing SasG in a dose-dependent fashion. These results suggest that SasG, Pls and AAP bind to the same as-yet-unidentified receptor on the surface of nasal epithelial cells. In addition, expression of SasG, like Pls, reduced adherence of S. aureus to fibronectin and fibrinogen. PMID- 14523110 TI - Identification and characterization of a novel autolysin (Aae) with adhesive properties from Staphylococcus epidermidis. AB - Staphylococcus epidermidis biofilm formation on polymer surfaces is considered a major pathogenicity factor in foreign-body-associated infections. Previously, the 148 kDa autolysin AtlE from S. epidermidis, which is involved in the initial attachment of the cells to polymer surfaces and also binds to the extracellular matrix protein vitronectin, was characterized. Here, the characterization of a novel autolysin/adhesin (Aae) in S. epidermidis is described. Aae was identified as a 35 kDa surface-associated protein that has bacteriolytic activity and binds vitronectin. Its N-terminal amino acid sequence was determined and the respective gene, aae, was cloned. DNA-sequence analysis revealed that aae encodes a deduced protein of 324 amino acids with a predicted molecular mass of 35 kDa. Aae contains three repetitive sequences in its N-terminal portion. These repeats comprise features of a putative peptidoglycan binding domain (LysM domain) found in a number of enzymes involved in cell-wall metabolism and also in some adhesins. Expression of aae by Escherichia coli and subsequent analysis revealed that Aae possesses bacteriolytic activity and adhesive properties. The interaction of Aae with fibrinogen, fibronectin and vitronectin was found to be dose-dependent and saturable and to occur with high affinity, by using the real time Biomolecular Interaction Analysis (BIA). Aae binds to the Aalpha- and Bbeta chains of fibrinogen and to the 29 kDa N-terminal fragment of fibronectin. In conclusion, Aae is a surface-associated protein with bacteriolytic and adhesive properties representing a new member of the staphylococcal autolysin/adhesins potentially involved in colonization. PMID- 14523111 TI - Identification and characterization of a fibronectin-binding protein from Clostridium difficile. AB - A 68 kDa fibronectin-binding protein (Fbp68) from Clostridium difficile displaying significant homology to several established or putative Fbps from other bacteria was identified. The one-copy gene is highly conserved in C. difficile isolates. Fbp68 was expressed in Escherichia coli in fusion with glutathione S-transferase; the fusion protein and the native Fbp68 were purified. Immunoblot analysis and cell fractionation experiments revealed that Fbp68 is present on the surface of the bacteria. Far-immuno dot-blotting demonstrated that Fbp68 was capable of fixing fibronectin. Indirect immunofluorescence and ELISA were employed to demonstrate that C. difficile could bind both soluble and immobilized fibronectin. With competitive adherence inhibition assays it was shown that antibodies raised against Fbp68 partially inhibited attachment of C. difficile to fibronectin and Vero cells. Furthermore, Vero cells could fix purified membrane-immobilized Fbp68. Thus Fbp68 appears to be one of the several adhesins identified to date in C. difficile. PMID- 14523112 TI - Alterations in the formation of lipopolysaccharide and membrane vesicles on the surface of Pseudomonas aeruginosa PAO1 under oxygen stress conditions. AB - It has been postulated that phenotypic variation in the relative expression of two chemically distinct types of lipopolysaccharide (LPS), a serotype-specific LPS (B-band) and a common antigen LPS (A-band) in Pseudomonas aeruginosa is an important mechanism enabling this opportunistic pathogen to alter its surface characteristics to mediate adhesion and to survive under extreme conditions. To further investigate this, the relative expression levels of the two distinct types of LPS in P. aeruginosa PAO1 were investigated with cells grown in a chemostat at different dissolved oxygen tensions (pO(2)). The A-band LPS was constitutively expressed as pO(2) was increased from nearly zero to 350 % of air saturation. In contrast, the B-band LPS showed a remarkable increase with increased pO(2). Almost no B-band LPS was found in cells grown at a pO(2) of less than 3 % of air saturation. Electron microscopic examination of cells revealed increased formation of membrane vesicles (MVs) on the surface of P. aeruginosa PAO1 under oxygen stress conditions. The toxicity of the supernatant of P. aeruginosa cultures to the growth of a hybridoma cell line significantly increased in samples taken from oxygen-stressed steady-state cultures. Furthermore, studies of adhesion in a continuous-flow biofilm culture revealed an increased adhesiveness for hydrophilic surfaces in P. aeruginosa PAO1 grown at a higher pO(2). The oxygen-dependent alterations of cell-surface components and properties observed in this work provide a possible explanation for the emergence of P. aeruginosa lacking the B-band LPS in chronically infected cystic fibrosis patients. The results are also useful for understanding the processes involved in the formation of MVs in P. aeruginosa. PMID- 14523113 TI - Proposed pathway for the biosynthesis of serovar-specific glycopeptidolipids in Mycobacterium avium serovar 2. AB - Members of the Mycobacterium avium complex are distinguished by the presence of highly antigenic surface molecules called glycopeptidolipids (GPLs) and the oligosaccharide portion of the serovar-specific GPL defines the 28 serovars. Previously, the genomic region (ser2) encoding the enzymes responsible for the glycosylation of the lipopeptide core to generate the serovar-2-specific GPLs has been described. In this work, the ser2 gene clusters of M. avium serovar 2 strains 2151 and TMC 724 were fully sequenced and compared to the homologous regions of M. avium serovar 1 strain 104, M. avium subsp. paratuberculosis and M. avium subsp. silvaticum. It was also determined that 104Rg, a mutant of strain 104 that produces truncated GPLs, lost several GPL biosynthesis genes by deletion. This comparison, together with analysis of protein similarities, supports a biosynthetic model in which serovar-2-specific GPLs are synthesized from a serovar-1-specific GPL intermediate that is derived from a non-specific GPL precursor. We also identified a gene encoding an enzyme that is necessary for the biosynthesis of serovar-3- and 9-specific GPLs, but not serovar-2-specific GPLs, suggesting that the different serovars may have evolved from the acquisition or loss of genetic information. In addition, a subcluster of genes for the biosynthesis and transfer of fucose, which are needed to make serovar specific GPLs such as those of serovar 2, is found in the non-GPL-producing M. avium subspecies paratuberculosis and silvaticum. PMID- 14523114 TI - Activation of hilA expression at low pH requires the signal sensor CpxA, but not the cognate response regulator CpxR, in Salmonella enterica serovar Typhimurium. AB - A two-component regulatory system, cpxR-cpxA, plays an important role in the pH dependent regulation of virF, a global activator for virulence determinants including invasion genes, in Shigella sonnei. The authors examined whether the cpxR-cpxA homologues have some function in the expression of Salmonella enterica serovar Typhimurium invasion genes via the regulation of hilA, an activator for these genes. In a Salmonella cpxA mutant, the hilA expression level was reduced to less than 10 % of that in the parent strain at pH 6.0. This mutant strain also showed undetectable synthesis of an invasion gene product, SipC, at pH 6.0 and reduced cell invasion capacity - as low as 20 % of that of the parent. In this mutant, the reduction in hilA expression was much less marked at pH 8.0 than at pH 6.0 - no less than 50 % of that in the parent, and no significant reduction was observed in either SipC synthesis or cell invasion rate, compared to the parent. Unexpectedly, a Salmonella cpxR mutant strain and the parent showed no apparent difference in all three characteristics described above at either pH. These results indicate that in Salmonella, the sensor kinase CpxA activates hilA, and consequently, invasion genes and cell invasion capacity at pH 6.0. At pH 8.0, however, CpxA does not seem to have a large role in activation of these factors. Further, the results show that this CpxA-mediated activation does not require its putative cognate response regulator, CpxR. This suggests that CpxA may interact with regulator(s) other than CpxR to achieve activation at low pH. PMID- 14523115 TI - Transcriptional regulation of drug efflux genes by EvgAS, a two-component system in Escherichia coli. AB - A constitutively active mutant of histidine kinase sensor EvgS was found to confer multi-drug resistance (MDR) to an acrA-deficient Escherichia coli, indicating the relationship between the two-component system EvgAS and the expression of the MDR system. The observed MDR also depended on an outer-membrane channel, TolC. Microarray and S1 mapping assays indicated that, in the presence of this constitutive mutant EvgS, the level of transcription increased for some MDR genes, including the drug efflux genes emrKY, yhiUV, acrAB, mdfA and tolC. Transcription in vitro of emrK increased by the addition of phosphorylated EvgA. Transcription activation of tolC by the activated EvgS was, however, dependent on both EvgAS and PhoPQ (Mg(2+)-responsive two-component system), in agreement with the presence of the binding site (PhoP box) for the regulator PhoP in the tolC promoter region. Transcription in vitro of yhiUV also appears to require an as yet-unidentified additional transcriptional factor besides EvgA. Taken together we propose that the expression of the MDR system is under a complex regulatory network, including the phosphorylated EvgA serving as the master regulator. PMID- 14523116 TI - A cryptic plasmid of Yersinia enterocolitica encodes a conjugative transfer system related to the regions of CloDF13 Mob and IncX Pil. AB - Yersinia enterocolitica 29930 (biotype 1A; O : 7,8), the producing strain of the phage-tail-like bacteriocin enterocoliticin, possesses a plasmid-encoded conjugative type IV transfer system. The genes of the conjugative system were found by screening of a cosmid library constructed from total DNA of strain 29930. The cosmid Cos100 consists of the vector SuperCos1 and an insert DNA of 40 303 bp derived from a cryptic plasmid of strain 29930. The conjugative transfer system consists of genes encoding a DNA transfer and replication system (Dtr) with close relationship to the mob region of the mobilizable plasmid CloDF13 and a gene cluster encoding a mating pair formation system (Mpf) closely related to the Mpf system of the IncX plasmid R6K. However, a gene encoding a homologue of TaxB, the coupling protein of the IncX system, is missing. The whole transfer region has a size of approximately 17 kb. The recombinant plasmid Cos100 was shown to be transferable between Escherichia coli and Yersinia with transfer frequencies up to 0.1 transconjugants per donor. Mutations generated by inserting a tetracycline cassette into putative tri genes yielded a transfer-deficient phenotype. Conjugative transfer of the cryptic plasmid could not be demonstrated in the original host Y. enterocolitica 29930. However, a kanamycin-resistance conferring derivative of the plasmid was successfully introduced into E. coli K 12 by transformation and was shown to be self-transmissible. Furthermore, Southern blot hybridization and PCR experiments were carried out to elucidate the distribution of the conjugative transfer system in Yersinia. In total, six Y. enterocolitica biotype 1A strains harbouring closely related systems on endogenous plasmids were identified. PMID- 14523117 TI - The curli biosynthesis regulator CsgD co-ordinates the expression of both positive and negative determinants for biofilm formation in Escherichia coli. AB - Production of curli, extracellular structures important for biofilm formation, is positively regulated by OmpR, which constitutes with the EnvZ protein an osmolarity-sensing two-component regulatory system. The expression of curli is cryptic in most Escherichia coli laboratory strains such as MG1655, due to the lack of csgD expression. The csgD gene encodes a transcription activator of the curli-subunit-encoding csgBA operon. The ompR234 up-mutation can restore csgD expression, resulting in curli production and increased biofilm formation. In this report, it is shown that ompR234-dependent csgD expression, in addition to csgBA activation during stationary phase of growth, stimulates expression of the yaiC gene and negatively regulates at least two other genes, pepD and yagS. The promoter regions of these four genes share a conserved 11 bp sequence (CGGGKGAKNKA), necessary for csgBA and yaiC regulation by CsgD. While at both the csgBA and yaiC promoters the sequence is located upstream of the promoter elements, in both yagS and pepD it overlaps either the putative -10 sequence or the transcription start point, suggesting that CsgD can function as both an activator and a repressor. Adhesion experiments show that csgD-independent expression of both yagS and pepD from a multicopy plasmid negatively affects biofilm formation, which, in contrast, is stimulated by yaiC expression. Thus it is proposed that CsgD stimulates biofilm formation in E. coli by contemporary activation of adhesion positive determinants (the curli-encoding csg operons and the product of the yaiC gene) and repression of negative effectors such as yagS and pepD. PMID- 14523118 TI - A three-dimensional, stochastic simulation of biofilm growth and transport related factors that affect structure. AB - Biofilm structural heterogeneity affects a broad range of microbially catalysed processes. Solute transport limitation and autoinhibitor production, two factors that contribute to heterogeneous biofilm development, were investigated using BacMIST, a computer simulation model. BacMIST combines a cellular automaton algorithm for biofilm growth with Brownian diffusion for solute transport. The simulation represented the growth of microbial unit cells in a three-dimensional domain modelled after a repeating section of a constant depth film fermenter. The simulation was implemented to analyse the effects of various levels of transport limitation on a growing single-species biofilm. In a system with rapid solute diffusion, cells throughout the biofilm grew at their maximum rate, and no solute gradient was formed over the biofilm thickness. In increasingly transport-limited systems, the rapidly growing fraction of the biofilm population decreased, and was found exclusively at the biofilm-liquid interface. Trans-biofilm growth substrate gradients also deepened with increasing transport limitation. Autoinhibitory biofilm growth was simulated for various rates of microbially produced inhibitor transport. Inhibitor transport rates affected both the biofilm population dynamics and the resulting biofilm structures. The formation of networks of void spaces in slow-growing regions of the biofilm and the development of columns in the fast-growing regions suggested a possible mechanism for the microscopically observed evolution of channels in biofilms. PMID- 14523119 TI - Aggregation of yeast cells: direct measurement of discrete lectin-carbohydrate interactions. AB - Aggregation of microbial cells mediated by specific interactions plays a pivotal role in the natural environment, in medicine and in biotechnological processes. Here we used atomic force microscopy (AFM) to measure individual lectin carbohydrate interactions involved in the flocculation of yeast cells, an aggregation event of crucial importance in fermentation technology. AFM probes functionalized with oligoglucose carbohydrates were used to record force-distance curves on living yeast cells at a rate of 0.5 micro m s(-1). Flocculating cells showed adhesion forces of 121+/-53 pN, reflecting the specific interaction between individual cell-surface lectins and glucose residues. Similar adhesion forces, 117+/-41 pN, were measured using probes functionalized with the lectin concanavalin A and attributed to specific binding to cell-surface mannose residues. By contrast, specific interaction forces were not observed in non flocculating conditions, i.e. in the presence of mannose or when using non flocculating cells, pointing to their involvement in yeast flocculation. The single molecule force spectroscopy measurements presented here provide a means to study a variety of cellular interactions at the molecular level, such as the adhesion of bacteria to animal and plant tissues. PMID- 14523120 TI - Degradation of alkanes and highly chlorinated benzenes, and production of biosurfactants, by a psychrophilic Rhodococcus sp. and genetic characterization of its chlorobenzene dioxygenase. AB - Rhodococcus sp. strain MS11 was isolated from a mixed culture. It displays a diverse range of metabolic capabilities. During growth on 1,2,4-trichlorobenzene, 1,2,4,5-tetrachlorobenzene (1,2,4,5-TeCB) and 3-chlorobenzoate stoichiometric amounts of chloride were released. It also utilized all three isomeric dichlorobenzenes and 1,2,3-trichlorobenzene as the sole carbon and energy source. Furthermore, the bacterium grew well on a great number of n-alkanes ranging from n-heptane to n-triacontane and on the branched alkane 2,6,10,14 tetramethylpentadecane (pristane) and slowly on n-hexane and n-pentatriacontane. It was able to grow at temperatures from 5 to 30 degrees C, with optimal growth at 20 degrees C, and could tolerate 6 % NaCl in mineral salts medium. Genes encoding the initial chlorobenzene dioxygenase were detected by using a primer pair that was designed against the alpha-subunit (TecA1) of the chlorobenzene dioxygenase of Ralstonia (formerly Burkholderia) sp. strain PS12. The amino acid sequence of the amplified part of the alpha-subunit of the chlorobenzene dioxygenase of Rhodococcus sp. strain MS11 showed >99 % identity to the alpha subunit of the chlorobenzene dioxygenase from Ralstonia sp. strain PS12 and the parts of both alpha-subunits responsible for substrate specificity were identical. The subsequent enzymes dihydrodiol dehydrogenase and chlorocatechol 1,2-dioxygenase were induced in cells grown on 1,2,4,5-TeCB. During cultivation on medium-chain-length n-alkanes ranging from n-decane to n-heptadecane, including 1-hexadecene, and on the branched alkane pristane, strain MS11 produced biosurfactants lowering the surface tension of the cultures from 72 to 2 mg per deciliter [176.8 micromol per liter]) who were undergoing coronary interventions. We randomly assigned them to either hemofiltration in an intensive care unit (ICU) (58 patients, with a mean [+/-SD] serum creatinine concentration of 3.0+/-1.0 mg per deciliter [265.2+/-88.4 micromol per liter]) or isotonic-saline hydration at a rate of 1 ml per kilogram of body weight per hour given in a step-down unit (56 patients, with a mean serum creatinine concentration of 3.1+/-1.0 mg per deciliter [274.0+/-88.4 micromol per liter]). Hemofiltration (fluid replacement rate, 1000 ml per hour without weight loss) and saline hydration were initiated 4 to 8 hours before the coronary intervention and were continued for 18 to 24 hours after the procedure was completed. RESULTS: An increase in the serum creatinine concentration of more than 25 percent from the base-line value after the coronary intervention occurred less frequently among the patients in the hemofiltration group than among the control patients (5 percent vs. 50 percent, P<0.001). Temporary renal-replacement therapy (hemodialysis or hemofiltration) was required in 25 percent of the control patients and in 3 percent of the patients in the hemofiltration group. The rate of in-hospital events was 9 percent in the hemofiltration group and 52 percent in the control group (P<0.001). In-hospital mortality was 2 percent in the hemofiltration group and 14 percent in the control group (P=0.02), and the cumulative one-year mortality was 10 percent and 30 percent, respectively (P=0.01). CONCLUSIONS: In patients with chronic renal failure who are undergoing percutaneous coronary interventions, periprocedural hemofiltration given in an ICU setting appears to be effective in preventing the deterioration of renal function due to contrast-agent-induced nephropathy and is associated with improved in-hospital and long-term outcomes. PMID- 14523142 TI - A trial of a 9-valent pneumococcal conjugate vaccine in children with and those without HIV infection. AB - BACKGROUND: Acute respiratory tract infections caused by Streptococcus pneumoniae are a leading cause of morbidity and mortality in young children. We evaluated the efficacy of a 9-valent pneumococcal conjugate vaccine in a randomized, double blind study in Soweto, South Africa. METHODS: At 6, 10, and 14 weeks of age, 19,922 children received the 9-valent pneumococcal polysaccharide vaccine conjugated to a noncatalytic cross-reacting mutant of diphtheria toxin (CRM197), and 19,914 received placebo. All children received Haemophilus influenzae type b conjugate vaccine. Efficacy and safety were analyzed according to the intention to-treat principle. RESULTS: Among children without human immunodeficiency virus (HIV) infection, the vaccine reduced the incidence of a first episode of invasive pneumococcal disease due to serotypes included in the vaccine by 83 percent (95 percent confidence interval, 39 to 97; 17 cases among controls and 3 among vaccine recipients). Among HIV-infected children, the efficacy was 65 percent (95 percent confidence interval, 24 to 86; 26 and 9 cases, respectively). Among children without HIV infection, the vaccine reduced the incidence of first episodes of radiologically confirmed alveolar consolidation by 20 percent (95 percent confidence interval, 2 to 35; 212 cases in the control group and 169 in the vaccinated group) in the intention-to-treat analysis and by 25 percent (95 percent confidence interval, 4 to 41; 158 and 119 cases, respectively) in the per protocol analysis (i.e., among fully vaccinated children). The incidence of invasive pneumococcal disease caused by penicillin-resistant strains was reduced by 67 percent (95 percent confidence interval, 19 to 88; 21 cases in the control group and 7 in the vaccinated group), and that caused by strains resistant to trimethoprim-sulfamethoxazole was reduced by 56 percent (95 percent confidence interval, 16 to 78; 32 and 14 cases, respectively). CONCLUSIONS: Vaccination with a 9-valent pneumococcal conjugate vaccine reduced the incidence of radiologically confirmed pneumonia. The vaccine also reduced the incidence of vaccine-serotype and antibiotic-resistant invasive pneumococcal disease among children with and those without HIV infection. PMID- 14523143 TI - Images in clinical medicine. Melanosis coli. PMID- 14523144 TI - Racial, ethnic, and geographic disparities in rates of knee arthroplasty among Medicare patients. AB - BACKGROUND: There are large variations in the use of knee arthroplasty among Medicare enrollees according to race or ethnic group and sex. Are racial and ethnic disparities more pronounced in some regions than in others, and if so, why? METHODS: We used all Medicare fee-for-service claims data for 1998 through 2000 to determine the incidence of knee arthroplasty according to Hospital Referral Region, sex, and race or ethnic group. A total of 430,726 knee arthroplasties were performed during the three-year study period. RESULTS: At the national level, the annual rate of knee arthroplasty was higher for non-Hispanic white women (5.97 procedures per 1000) than for Hispanic women (5.37 per 1000) and black women (4.84 per 1000). The rate for non-Hispanic white men (4.82 procedures per 1000) was higher than that for Hispanic men (3.46 per 1000) and more than double that for black men (1.84 per 1000). The rates were significantly lower for black men than for non-Hispanic white men in nearly every region of the country (P<0.05). For the Hispanic population and for black women, racial or ethnic disparities at the national level were due in part to geographic differences rather than to differences in the rates for different racial and ethnic groups within geographic areas. Residential segregation and low income levels contributed to racial and ethnic disparities in arthroplasty rates. CONCLUSIONS: In the Medicare population, the rate of surgical treatment for osteoarthritis of the knee varies dramatically according to sex, race or ethnic group, and region. These variations underscore the importance of geography and sex in determining racial or ethnic barriers to health care. PMID- 14523145 TI - Chronic constipation. PMID- 14523146 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 31-2003. A 44-year-old man with HIV infection and a right atrial mass. PMID- 14523147 TI - Racial disparities--the need for research and action. PMID- 14523148 TI - The 50th anniversary of the discovery of immunologic tolerance. PMID- 14523149 TI - Treatment of bronchiolitis. PMID- 14523150 TI - Dalteparin compared with an oral anticoagulant for thromboprophylaxis in patients with cancer. PMID- 14523151 TI - Cardiovascular genomics. PMID- 14523152 TI - Valvular heart disease in pregnancy. PMID- 14523153 TI - Cardiac transplantation in an HIV-1-infected patient. PMID- 14523154 TI - Complex umbilical-cord knot. PMID- 14523155 TI - Persistent efficacy of Vi conjugate vaccine against typhoid fever in young children. PMID- 14523156 TI - Images in clinical medicine. Kala-azar. PMID- 14523157 TI - Genomic background drives the divergence of duplicated amylase genes at synonymous sites in Drosophila. AB - In some Drosophila species, there are two types of greatly diverged amylase (Amy) genes (Amy clusters 1 and 2), each encoding active amylase isozymes. Cluster 1 is located at the middle of its chromosomal arm, and the region has a normal local recombination rate. However, cluster 2 is near the centromere, and this region is known to have a reduced recombination rate. Although nonsynonymous substitutions follow a molecular clock, synonymous substitutions were accelerated in cluster 2 after gene duplications. This resulted in a higher GC content at the third codon position (GC3) and codon usage bias in cluster 1, and lower GC3 content and codon usage bias in the cluster 2. However, no systematic difference in GC content was observed in the first and second codon positions or the 3'-flanking regions. Therefore, differences in local recombination rate rather than mutation bias might explain the divergence at synonymous sites between the two Amy clusters within species (Hill-Robertson effect). Alternatively, the different patterns and levels of expression between the two clusters may imply that the reduced expression level in cluster 2 caused by chromatin potentiation decreased the codon bias. Both of these hypotheses imply the importance of the genomic background as a driving force of divergence between non-tandemly duplicated genes. PMID- 14523158 TI - Sequence determinants of quaternary structure in lumazine synthase. AB - Riboflavin, an essential cofactor for all organisms, is biosynthesized in plants, fungi and microorganisms. The penultimate step in the pathway is catalyzed by the enzyme lumazine synthase. One of the most distinctive characteristics of this enzyme is that it is found in different species in two different quaternary structures, pentameric and icosahedral, built from practically the same structural monomeric unit. In fact, the icosahedral structure is best described as a capsid of twelve pentamers. Despite this noticeable difference, the active sites are virtually identical in all structurally studied members. Furthermore, the main regions involved in the catalysis are located at the interface between adjacent subunits in the pentamer. Thus, the two quaternary forms of the enzyme must meet similar structural requirements to achieve their function, but, at the same time, they should differ in the sequence traits responsible for the different quaternary structures observed. Here, we present a combined analysis that includes sequence-structure and evolutionary studies to find the sequence determinants of the different quaternary assemblies of this enzyme. A data set containing 86 sequences of the lumazine synthase family was recovered by sequence similarity searches. Seven of them had resolved three-dimensional structures. A subsequent phylogenetic reconstruction by maximum parsimony (MP) allowed division of the total set into two clusters in accord with their quaternary structure. The comparison between the patterns of three-dimensional contacts derived from the known three-dimensional structures and variation in sequence conservation revealed a significant shift in structural constraints of certain positions. Also, to explore the changes in functional constraints between the two groups, site-specific evolutionary rate shifts were analyzed. We found that the positions involved in icosahedral contacts suffer a larger increase in constraints than the rest. We found eight sequence sites that would be the most important icosahedral sequence determinants. We discuss our results and compare them with previous work. These findings should contribute to refinement of the current structural data, to the design of assays that explore the role of these positions, to the structural characterization of new sequences, and to initiation of a study of the underlying evolutionary mechanisms. PMID- 14523159 TI - Fluoride. PMID- 14523160 TI - What's new in the management of pain in children. PMID- 14523161 TI - Index of suspicion. PMID- 14523162 TI - Question from the clinician: tuberculosis prophylaxis. PMID- 14523163 TI - Children and adolescents who have schizophrenia. PMID- 14523164 TI - Enteroviruses. PMID- 14523165 TI - Association between peak serum bilirubin and neurodevelopmental outcomes in extremely low birth weight infants. AB - OBJECTIVE: To assess the association between peak total serum bilirubin (PSB) levels during the first 2 weeks of life and neurodevelopmental outcomes of extremely low birth weight (ELBW) infants at 18 to 22 months' postmenstrual age. METHODS: A retrospective analysis was conducted of a cohort of ELBW infants (401 1000 g) who survived to 14 days of age in the 12 participating centers of the National Institute of Child Health and Human Development Neonatal Research Network between January 1, 1994, and December 31, 1997. Demographic and clinical risk factors and PSB levels during the first 14 days were analyzed with reference to death or adverse neurodevelopmental outcomes at 18 to 22 months' postmenstrual age. The neurodevelopmental variables considered were Psychomotor Developmental Index (PDI) <70, Mental Developmental Index (MDI) <70, moderate or severe cerebral palsy (CP), hearing impairment (needs hearing aids), and a composite category designated as neurodevelopmental impairment (NDI). The NDI is defined as infants with any 1 or more of the following: PDI <70, MDI <70, moderate to severe CP, bilateral blindness, or bilateral hearing impairment requiring amplification. RESULTS: The subjects of this cohort analysis are infants who were admitted to the Network centers during calendar years 1994-1997 and survived beyond 14 days and had PSB recorded during the 14-day period. From this cohort, 3246 infants survived at discharge, 79 died after discharge, and 592 were lost to follow-up. Thus, 2575 of 3167 infants were seen in the follow-up clinics with a compliance rate of 81%. Logistic regression analysis showed that various demographic and clinical variables are associated with poor neurodevelopmental outcomes. After adjustment for these risk factor, significant association were found between PSB (mg/dL) and death or NDI (odds ratio: 1.068; 95% confidence interval [CI]: 1.03 1.11); PDI <70 (R = 1.057; 95% CI: 1.00-1.12), and hearing impairment requiring hearing aids (odds ratio: 1138; 95% CI: 1.00-1.30). There was no significant association between PSB (mg/dL) and CP, MDI <70, and NDI. CONCLUSIONS: PSB concentrations during the first 2 weeks of life are directly correlated with death or NDI, hearing impairment, and PDI <70 in ELBW infants. The statistical association based on retrospective analysis of observational data and relatively small effect size should be interpreted with caution. Furthermore, because of the possibility of compounding effects of variables on outcome, the potential benefits of moderate hyperbilirubinemia and the potential adverse effects of phototherapy, a randomized, controlled trial of aggressive and conservative phototherapy is needed to address this controversial issue. PMID- 14523166 TI - Accuracy of analog telephonic stethoscopy for pediatric telecardiology. AB - OBJECTIVE: Wide-bandwidth electronic stethoscopy is reliable and accurate for pediatric telecardiology. We tested a much less expensive and more convenient system for the same purpose, a narrow-bandwidth telephonic stethoscope (TS). METHODS: Seventy-six consecutive patients (mean age: 10.0; standard deviation: 6.5 years) in a pediatric cardiology outpatient clinic were studied. One pediatric cardiologist examined the patients with his acoustic stethoscope (AS); a second examined them within a few minutes using a remote TS. A nurse placed the TS chest piece as directed by the remote examiner via intercom, but neither video examination nor conversation with the parent/patient were permitted. Examiners independently recorded the stethoscope findings for all heart sounds, all murmurs, and heart disease (present/absent). TS accuracy was indexed using the kappa statistic for TS/AS agreement and for TS agreement with auscultatory findings predicted from echocardiographic (echo) studies (N = 49). RESULTS: TS/AS agreement was satisfactory for presence/absence of heart disease (kappa = 0.63) and for organic, functional, vibratory, diastolic aortic, and diastolic pulmonic murmurs (kappa range: 0.65-0.75). For other specific murmurs and all heart sounds, TS/AS agreement was either unsatisfactory (kappa < or = 0.60) or indeterminate because prevalence was 0. TS-AS agreement improved when the TS was used by the more-experienced TS examiner and with patients at least 5 years of age. When the older children were examined by the more TS-experienced examiner, the TS-echo comparison yielded kappa = 0.90, raw agreement = 0.96, sensitivity = 0.94, and specificity = 1.00. CONCLUSIONS: In pediatric patients, a narrow bandwidth telephonic stethoscope can accurately distinguish between functional and organic murmurs and thus can detect heart disease. Accuracy is greatest when the instrument is used by an experienced examiner with patients at least 5 years of age. PMID- 14523167 TI - Pediatric training and job market trends: results from the American Academy of Pediatrics third-year resident survey, 1997-2002. AB - OBJECTIVES: To examine trends in pediatric residents' training and job search experiences from 1997-2002. METHODS: Annual national random samples of 500 graduating pediatric residents were surveyed, and responses were compared across survey years to identify trends. The overall response rate was 71%. RESULTS: From 1997-2002, there were more female residents and US underrepresented minorities and fewer international medical graduates. Each successive group of residents rated higher their preparation for fellowship training, for child advocacy, and for assessing community needs. These increases paralleled an increase in resident exposure to community sites as part of their residency education. Educational debt (in 2002 dollars) for residents increased substantially across survey years from an average of 64 070 dollars in 1997 to 87 539 dollars in 2002. Meanwhile, starting salaries (in 2002 dollars) for residents entering general pediatrics actually decreased. Interest in general pediatrics among residents decreased, whereas interest in subspecialty practice increased during this time period. Fewer residents with general pediatrics as a career goal had a job when surveyed, and fewer obtained their first-choice positions across years. CONCLUSIONS: Experiences of graduating residents over the past 6 years provide insights into changes in pediatric residency education and the pediatric workforce. Efforts by pediatric educators and academic leaders to increase community experiences and child advocacy and to encourage greater interest in pediatric subspecialty careers seem to be succeeding. Unfortunately, demand for general pediatricians is weakening, and residents are experiencing increasing debt burdens. PMID- 14523168 TI - Early reversal of pediatric-neonatal septic shock by community physicians is associated with improved outcome. AB - OBJECTIVE: Experimental and clinical studies of septic shock support the concept that early resuscitation with fluid and inotropic therapies improves survival in a time-dependent manner. The new American College of Critical Care Medicine Pediatric Advanced Life Support (ACCM-PALS) Guidelines for hemodynamic support of newborns and children in septic shock recommend this therapeutic approach. The objective of this study was to determine whether early septic shock reversal and use of resuscitation practice consistent with the new ACCM-PALS Guidelines by community physicians is associated with improved outcome. METHODS: A 9-year (January 1993-December 2001) retrospective cohort study was conducted of 91 infants and children who presented to local community hospitals with septic shock and required transport to Children's Hospital of Pittsburgh. Shock reversal (defined by return of normal systolic blood pressure and capillary refill time), resuscitation practice concurrence with ACCM-PALS Guidelines, and hospital mortality were measured. RESULTS: Overall, 26 (29%) patients died. Community physicians successfully achieved shock reversal in 24 (26%) patients at a median time of 75 minutes (when the transport team arrived at the patient's bedside), which was associated with 96% survival and >9-fold increased odds of survival (9.49 [1.07-83.89]). Each additional hour of persistent shock was associated with >2-fold increased odds of mortality (2.29 [1.19-4.44]). Nonsurvivors, compared with survivors, were treated with more inotropic therapies (dopamine/dobutamine [42% vs 20%] and epinephrine/norepinephrine [42% vs 6%]) but not increased fluid therapy (median volume; 32.9 mL/kg vs 20.0 mL/kg). Resuscitation practice was consistent with ACCM-PALS Guidelines in only 27 (30%) patients; however, when practice was in agreement with guideline recommendations, a lower mortality was observed (8% vs 38%). CONCLUSIONS: Early recognition and aggressive resuscitation of pediatric-neonatal septic shock by community physicians can save lives. Educational programs that promote ACCM-PALS recommended rapid, stepwise escalations in fluid as well as inotropic therapies may have value in improving outcomes in these children. PMID- 14523169 TI - Is interleukin-6 -174 genotype associated with the development of septicemia in preterm infants? AB - OBJECTIVE: Systemic infection affects one quarter of preterm infants. Defense from infection is in part mediated by the cytokine interleukin-6 (IL-6). We tested the hypothesis that the IL-6 -174 GG genotype, associated with lower IL-6 response to inflammation, is also associated with the development of septicemia in preterm infants. METHODS: The study group comprised 157 infants who were born at < or =32 weeks. Genotype distribution (34% [54] GG, 46% [72] GC, 20% [31] CC) and C allele frequency (0.43; 95% confidence interval [CI]: 0.37-0.48) were similar to the UK adult population. Among the patients who developed bacterially confirmed septicemia (n = 51 [33%]), there was a significantly higher prevalence of the IL-6 -174 GG genotype than that observed in those who did not develop infection (47% vs 28% for GG: odds ratio [OR]: 2.3; 95% CI: 1.1-4.5). This association remained statistically significant (OR: 2.7; 95% CI: 1.2-6.3) after multiple binary logistic regression adjustment for other significant predictors of the development of septicemia. Late infection alone was similarly associated with GG genotype (septicemia 47% vs no septicemia 29% for GG: OR: 2.2; 95% CI: 1.1-4.3). CONCLUSIONS: Variation in the IL-6 gene seems to influence the defense against bacterial pathogens in the very preterm infant. PMID- 14523170 TI - Dating of bruises in children: an assessment of physician accuracy. AB - OBJECTIVE: To determine whether physicians can estimate accurately the age of an accidental bruise on direct physical examination. METHODS: Children who presented to the emergency department of a children's hospital with accidental bruises of known age and origin had demographic data and information about their injury recorded. History-blinded emergency pediatricians, other physicians, and trainees (fellows, residents, and medical students) independently examined the bruised area and recorded injury characteristics and age estimation and ranked characteristics that influenced their estimation. RESULTS: Fifty children with accidental bruises were enrolled. Emergency pediatricians' accuracy of age estimation within 24 hours of actual age was 47.6%. Individual emergency pediatrician's accuracy ranged from 0% to 100%, and the interobserver reliability was poor (kappa = -0.03). Accuracy within 24 hours of actual age was 29.4% for other physicians and 36.8% for trainees, which was similar to the emergency pediatricians. Observers reported using color primarily to estimate age, followed by tenderness and then swelling; however, none of these factors was significantly correlated with accuracy. CONCLUSIONS: Physician estimates of bruise age are highly inaccurate within 24 hours of the actual age of the injury. Large individual variability and poor interrater reliability also suggest that caution must be used when interpreting these estimates. This study supports earlier studies, urging extreme caution in estimating bruise age, even when such estimates are based on direct examination of the injured area. PMID- 14523171 TI - Neuroimaging of intraparenchymal lesions predicts outcome in shaken baby syndrome. AB - OBJECTIVE: Studies of long-term outcome on nonaccidental head injury (NAHI) in young children have shown severe neurodevelopmental sequelae in most cases. For improving the knowledge of outcome and for identifying prognostic factors, additional clinical and cerebral imaging data are needed. The aim of this study was to describe clinical and imaging features over time and to consider their value for predicting neurodevelopmental outcome. METHODS: A retrospective medical record review was conducted of 23 children with confirmed NAHI, for whom an extended follow-up of 2.5 to 13 years (mean: 6 years) was contemplated. Glasgow Coma Scale scores, severity of retinal hemorrhages, presence of skull fractures, cranial growth deceleration, and sequential neuroimaging data (computed tomography and/or magnetic resonance imaging) were compared with patterns of clinical evolution assessed by the Glasgow Outcome Scale. RESULTS: Clinical outcome showed that 14 (61%) children had severe disabilities, 8 (35%) had moderate disabilities, and 1 (4%) was normal. A low initial Glasgow Coma Scale score, severe retinal hemorrhages, presence of skull fracture, and cranial growth deceleration were significantly associated with poor developmental outcome. Eighteen of the 23 patients had abnormal magnetic resonance imaging scans. This examination disclosed atrophy when performed beyond 15 days of injury. Atrophy seemingly resulted from various brain lesions, namely, contusions, infarcts, and other lesions within the white matter. Presence of intraparenchymal brain lesions within the first 3 months was significantly associated with neurodevelopmental impairment. Severity of motor and cognitive dysfunctions was related to the extent of intraparenchymal lesions. CONCLUSIONS: Early clinical and radiologic findings in NAHI are of prognostic value for neurodevelopmental outcome. PMID- 14523172 TI - Risk of anaphylaxis after vaccination of children and adolescents. AB - OBJECTIVE: To quantify the risk of anaphylaxis after vaccination of children and adolescents. METHODS: The study population consisted of children and adolescents who were enrolled at 4 health maintenance organizations that participated in the Vaccine Safety Datalink Project. For the period 1991-1997, we identified potential cases by searching for occurrences of International Classification of Diseases, Ninth Revision (ICD-9) code 995.0 (anaphylactic shock), E948.0 through E948.9 (adverse reaction from bacterial vaccines), and E949.0 through E949.9 (adverse reaction from other vaccines and biological substances). At 1 study site, we also included a range of other allergy codes. We restricted to diagnoses on days 0 to 2 after vaccination (ICD-9 995.0) or day 0 (all other ICD-9 codes). We then reviewed the medical record to confirm the diagnosis. RESULTS: We identified 5 cases of potentially vaccine-associated anaphylaxis after administration of 7 644 049 vaccine doses, for a risk of 0.65 cases/million doses (95% confidence interval: 0.21-1.53). None of the episodes resulted in death. Vaccines that were administered before the anaphylactic episodes were generally given in combination and included measles-mumps-rubella, hepatitis B, diphtheria tetanus, diphtheria-tetanus-pertussis, Haemophilus influenzae type b, and oral polio vaccine. One case of anaphylaxis followed measles-mumps-rubella vaccine alone. At the site at which we reviewed additional allergy codes, we identified 1 case after 653 990 vaccine doses, for a risk of 1.53 cases/million doses (95% confidence interval: 0.04-8.52). CONCLUSIONS: Patients and health care providers can be reassured that vaccine-associated anaphylaxis is a rare event. Nevertheless, providers should be prepared to provide immediate medical treatment should it occur. PMID- 14523173 TI - Potential burden of universal influenza vaccination of young children on visits to primary care practices. AB - OBJECTIVE: To estimate the additional number of visits to primary care practices that would be required to deliver universal influenza vaccination to 6- to 23 month-old children. METHODS: Children who were covered by commercial and Medicaid managed care plans (70% of children in the region; >8000 children in each of 3 consecutive influenza seasons) in the 6-county region surrounding and including Rochester, New York, were studied. An analysis was conducted of insurance claims for visits (well-child care [WCC]; all other visits) to primary care practices during 3 consecutive influenza vaccination seasons (1998-2001). We determined the proportion of children who made 1 or 2 visits during the potential influenza vaccination period, simulating several possible lengths of time available for influenza vaccination (2, 3, 4, or 5 months). We measured the proportion of children who were vaccinated during each influenza vaccination period. The added visit burden was defined as the number of additional visits that would be required to vaccinate all children, simulating 2 scenarios: 1) administering influenza vaccination only during WCC visits and 2) considering all visits as opportunities for influenza vaccination. RESULTS: Results were similar for each influenza season. Considering a 3-month influenza vaccination window and assuming that no opportunities were missed, if only WCC visits were used for influenza vaccination, then 74% of 6- to 23-month-olds would require at least 1 additional visit for vaccination--39% would require 1 additional visit and 35% would require 2 additional visits. If all visits to the practice were used for influenza vaccination during the 3-month window, then 46% would require at least 1 additional visit--34% would require 1 additional visit and 12% would require 2 additional visits. Longer vaccination periods would require fewer additional visits; eg, if a 4-month period were available, then 54% of children would require 1 or 2 additional visits if only WCC visits were used and 29% would require 1 or 2 additional visits if all visits were used for influenza vaccinations. Younger children (eg, 6- to 11-month-olds) would require fewer additional visits than older children (12- to 23-month-olds) because younger children already have more visits to primary care practices. CONCLUSIONS: Implementation of universal influenza vaccination will result in a substantial increased burden to primary care practices in terms of additional visits for influenza vaccination. Practice-level strategies to minimize the additional burden include 1) using all visits (not just WCC visits) as opportunities for vaccination, 2) providing influenza vaccination for the maximum possible time period by starting to vaccinate as early as possible and continuing to vaccinate as late as possible, and 3) implementing short and efficient vaccination-only visits to accommodate the many additional visits to the practice. PMID- 14523174 TI - Healing patterns in anogenital injuries: a longitudinal study of injuries associated with sexual abuse, accidental injuries, or genital surgery in the preadolescent child. AB - OBJECTIVE: To study healing patterns of anogenital trauma in prepubescent children. METHODS: A prospective 10-year study was conducted of 94 children who had anogenital trauma and were followed to healing and documented using a colposcope with 35-mm camera attachment. RESULTS: The 13 boys and 81 girls were referred with injuries as a result of sexual assault or anogenital trauma. Hymenal injuries occurred in 37 cases; 2 transections healed after surgery, and 15 persisted unchanged. Partial tears, hymenal abrasions, or hematomas healed completely or with minor nonspecific changes. Of the 47 injuries to the posterior fourchette, 22 abrasions, hematomas, or tears healed completely; 12 tears healed with vascular changes; 2 developed labial fusions; 10 lacerations required surgery; and 6 scarred and 4 healed with vascular changes. Only 2 of 39 cases of perihymenal trauma healed with vascular changes. All 17 cases of labial trauma healed completely. Anal trauma healed completely in 29 of 31 with scarring occurring in only 2 cases that required surgery. CONCLUSIONS: Anogenital trauma heals quickly, often without residua. Of the 94 cases, there were diagnostic anatomic changes in the 15 cases of hymenal transections (2 other cases healed completely with surgical reconstruction), 6 cases after surgical repair of posterior fourchette, and 2 cases of anal scarring after surgery. PMID- 14523175 TI - Hemodynamic changes in anemic premature infants: are we allowing the hematocrits to fall too low? AB - OBJECTIVE: Currently, many nurseries allow hematocrits to fall to <21% in apparently "stable" premature infants before considering a blood transfusion. We evaluated clinical changes and hemodynamic changes by echocardiogram in "stable" anemic premature infants before, during, and after transfusion. METHODS: "Stable" premature infants (< or =32 weeks' gestation) who were to receive transfusions (2 aliquots of 10 mL/kg packed red blood cells, 12 hours apart) were eligible for prospective enrollment. Cardiac function by echocardiography and vital signs were measured 4 times: 1 to 3 hours before and 2 to 4 hours after the initial aliquot and 4 to 7 hours and 27 to 34 hours after the second aliquot. Infants were grouped prospectively according to pretransfusion hematocrit ranges for analysis: < or =21% (low), 22% to 26% (mid), and > or =27% (high). RESULTS: Thirty-two infants were enrolled. No differences were observed between the groups in sex, birth weight, postconceptional age, or postnatal weight at enrollment. Before transfusion, low- and mid-range groups had higher left ventricular end systolic and diastolic diameters, in comparison with high range. Low range had increased stroke volume in comparison with the high-range group. These changes persisted after transfusion. Mean diastolic blood pressure rose and peak velocity in the aorta fell in the low-range group after transfusion. Pretransfusion hematocrit was correlated with but poorly predictive of echocardiographic measurements. Infants with inappropriate weight gain had increased ventricular end diastolic diameters, consistent with congestive heart failure. CONCLUSIONS: Apparently "stable" anemic premature infants may be in a clinically unrecognized high cardiac output state, and some echocardiographic measurements do not improve within 48 hours after transfusion. The benefits of transfusion practices guided by measures of cardiac function should be evaluated. PMID- 14523176 TI - Behavioral and developmental effects of preventing iron-deficiency anemia in healthy full-term infants. AB - OBJECTIVE: To determine the behavioral and developmental effects of preventing iron-deficiency anemia in infancy. METHODS: Healthy full-term Chilean infants who were free of iron-deficiency anemia at 6 months were assigned to high- or low iron groups or to high- or no-added-iron groups. Behavioral/developmental outcomes at 12 months of age included overall mental and motor test scores and specific measures of motor functioning, cognitive processing, and behavior. There were no differences between high- and low-iron groups in the prevalence of iron deficiency anemia or behavioral/developmental outcome, and they were combined to form an iron-supplemented group (n = 1123) for comparison with the no-added-iron group (n = 534). RESULTS: At 12 months, iron-deficiency anemia was present in 3.1% and 22.6% of the supplemented and unsupplemented groups, respectively. The groups differed in specific behavioral/developmental outcomes but not on global test scores. Infants who did not receive supplemental iron processed information slower. They were less likely to show positive affect, interact socially, or check their caregivers' reactions. A smaller proportion of them resisted giving up toys and test materials, and more could not be soothed by words or objects when upset. They crawled somewhat later and were more likely to be tremulous. CONCLUSIONS: The results suggest that unsupplemented infants responded less positively to the physical and social environment. The observed differences seem to be congruent with current understanding of the effects of iron deficiency on the developing brain. The study shows that healthy full-term infants may receive developmental and behavioral benefits from iron supplementation in the first year of life. PMID- 14523177 TI - Cerebral function monitoring: a new scoring system for the evaluation of brain maturation in neonates. AB - OBJECTIVE: Cerebral function monitoring (CFM), using compressed single-channel amplitude-integrated electroencephalogram recorded from 2 biparietal electrodes, has been shown previously to be a simple bedside tool for monitoring neonatal central nervous system (CNS) status. As the pattern of the CFM changes with gestational age, the technique can be used to assess brain maturation in premature infants. We have developed a new scoring system for the interpretation of neonatal CFM recordings. The objective of this study was to evaluate CFM tracings at increasing gestational and postnatal ages to develop a scoring system to quantify CFM pattern changes. METHODS: Term and preterm neonates were studied with CFM at 12 to 24 hours of life, 48 to 72 hours of life, and then weekly or biweekly until hospital discharge. Each study comprised 8 to 24 hours of continuous CFM recording. CFM recordings were evaluated using the scoring system for record continuity, presence of cyclic changes in electrical activity, degree of voltage amplitude depression, and bandwidth. Each variable was scored for each recording. All variables were summed to yield a total score (minimum 0, maximum 13). Total scores were correlated with gestational and postconceptional ages. RESULTS: Thirty infants were studied with gestational ages at birth that ranged from 24 to 39 weeks and birth weights that varied between 450 and 3850 g. A total of 146 CFM tracings were analyzed. With advancing gestational and postconceptional age, scores for each variable as well as total scores progressively increased with CNS maturation. The highest scores were attained at 35 to 36 weeks' postconceptional age, which corresponded to previously reported subjective observations performed by visual description of CFM patterns. Of the 4 component variables that we analyzed, the most sensitive indicators of CNS maturity were 1) the presence of a cycling pattern, 2) the continuity of the record pattern, and 3) the CFM recording bandwidth. CONCLUSIONS: Our proposed scoring system may be a valuable tool to quantify changes during CFM more objectively, reflecting variations in CNS activity in newborn infants and allowing for better statistical comparisons between amplitude-integrated electroencephalogram tracings from different patients as well as from the same patient at different points of time. PMID- 14523178 TI - Antibiotic-resistant Streptococcus pneumoniae in the heptavalent pneumococcal conjugate vaccine era: predictors of carriage in a multicommunity sample. AB - OBJECTIVE: Despite immunization with heptavalent pneumococcal conjugate vaccine (PCV7), the rising prevalence of antibiotic resistance makes Streptococcus pneumoniae a continuing threat to child health. Data on carriage of resistant organisms by healthy children in communities in which immunization with PCV7 has been implemented will help to define and decrease these risks further. METHODS: Children who were <7 years old, resided in a study community, and presented for routine well care or a "sick" visit between March 13 and May 11, 2001, at 31 primary care practices in 16 geographically distinct Massachusetts communities were studied. Consenting parents provided demographic information and data on potential risk factors for carriage of S pneumoniae and of penicillin nonsusceptible S pneumoniae (PNSP). S pneumoniae isolates from nasopharyngeal specimens were tested for resistance to commonly used antibiotics including penicillin, ceftriaxone, erythromycin, and trimethoprim/sulfamethoxazole. Isolates were serotyped and grouped into PCV7-included serotypes, potentially cross-reactive serotypes (ie, an organism of a serogroup included in the vaccine), or non-PCV7 serotypes. Diagnosis on the day of collection, history of recent antibiotic use, and history of PCV7 immunization were obtained by chart review. Separate bivariate and multivariate analyses were performed to identify correlates of colonization with S pneumoniae and colonization with PNSP, accounting for clustering within communities. RESULTS: S pneumoniae was isolated from the nasopharynx of 190 (26%) of the 742 children studied. Of the 166 tested, 33% were nonsusceptible to penicillin, with 14% showing intermediate susceptibility (minimum inhibitory concentration [MIC] 0.12-1.0) and 19% fully resistant (MIC > or =2). Nonsusceptibility to other antibiotics was common, including ceftriaxone (14%), erythromycin (22%), and trimethoprim/sulfa (31%); 20% of S pneumoniae isolates were not susceptible to > or =3 antibiotics. Thirty six percent of isolates were of serotypes covered by PCV7; 30% were of PCV7 serogroups and potentially cross-reactive, but not 1 of the 7 included serotypes; and 34% were unrelated to PCV7 serogroups. Nonsusceptibility to penicillin was more common in PCV7-included strains (45%) and potentially cross-reactive strains (51%) than in non-PCV7 serotypes (8%). Risk factors for PNSP colonization included child care attendance (odds ratio [OR]: 3.9; 95% confidence interval [CI]: 2.3-6.5), current respiratory tract infection (OR: 4.7; 95% CI: 2.5-8.6), and recent antibiotic use (OR: 1.7; 95% CI: 1.0-2.8). PCV7 immunization was associated with decreased carriage of PCV7-included serotypes but not with an overall decrease in S pneumoniae colonization or with a decline in PNSP colonization. CONCLUSIONS: In this multicommunity sample, pneumococcal antibiotic resistance was common and was most frequently found in PCV7-included and PCV7 serogroup strains. The long-term impact of PCV7 immunization will be partially determined by the protection that it affords against invasive infection with potentially cross-reactive serotypes, as well as the virulence and future resistance patterns of unrelated serotypes. PMID- 14523179 TI - Symptoms of sleep-disordered breathing in 5-year-old children are associated with sleepiness and problem behaviors. AB - OBJECTIVE: Sleep-disordered breathing (SDB) in children is reportedly associated with problem behaviors suggestive of attention-deficit/hyperactivity disorder; however, there are few data on the relation of SDB to problem behaviors in the general pediatric population. The goal of this study was to assess the prevalence of SDB symptoms in 5-year-old children and their relation to sleepiness and problem behaviors. METHODS: A population-based, cross-sectional survey was conducted of a birth cohort of children who were born in eastern Massachusetts. Subjects were 3019 5-year-old children (1551 boys, 1468 girls) who were enrolled in the Infant Care Practices Study and whose mothers were contacted within 3 months of their child's fifth birthday. A parent-completed questionnaire was used to ascertain the presence and intensity of snoring and other SDB symptoms and the presence of daytime sleepiness and problem behaviors. Parent-reported hyperactivity, inattention, and aggressiveness were each assessed by a single question that was validated against the Conners' Parent Rating Scale. SDB was defined as frequent or loud snoring; trouble breathing or loud, noisy breathing during sleep; or witnessed sleep apnea. RESULTS: Parent-reported hyperactivity (19%) and inattention (18%) were common, with aggressiveness (12%) and daytime sleepiness (10%) reported somewhat less often. SDB symptoms were present in 744 (25%) children. Compared with children without snoring or other symptoms of SDB, children with SDB symptoms were significantly more likely to have parent-reported daytime sleepiness (odds ratio [OR]: 2.2; 95% confidence interval [CI]: 1.7-2.8) and problem behaviors, including hyperactivity (OR: 2.5; CI: 2.0-3.0), inattention (OR: 2.1; 95% CI: 1.7-2.6), and aggressiveness (OR: 2.1; 95% CI: 1.6 2.6). These associations remained significant after adjustment for sex, race/ethnicity, maternal education level, maternal marital status, household income, and respiratory health history. CONCLUSIONS: SDB symptoms are common in 5 year-old children and are associated with an increased risk of daytime sleepiness and with problem behaviors suggestive of attention-deficit/hyperactivity disorder. PMID- 14523180 TI - Back to sleep: can we influence child care providers? AB - OBJECTIVE: Despite the fact that 20% of sudden infant death syndrome (SIDS) deaths occur in child care settings, many child care providers continue to be unaware of the association of SIDS and infant sleep position and/or are misinformed as to the risks and benefits of the various sleep positions. The objective of this study was to determine whether an educational program for child care providers regarding SIDS and safe sleep environment is effective in 1) providing basic information and understanding regarding SIDS risk reduction practices, 2) changing child care provider behavior, and 3) promoting development of written sleep position policies. METHODS: We designed a 60-minute educational in-service for child care providers, to be led by a trained health educator. All providers who attended the in-service were asked to complete surveys before and after the in-service. Surveys assessed provider knowledge, beliefs, and practices. A 6-month follow-up interview was conducted with child care centers that had providers participating in the in-service. RESULTS: A total of 96 child care providers attended the educational in-service. Providers who were using the supine position exclusively increased from 44.8% to 78.1%. This change in behavior was sustained, with 85% of centers placing infants exclusively supine 6 months after the intervention. Awareness of the American Academy of Pediatrics recommendation of supine as the preferred position for infants increased from 47.9% to 78.1%, and 67.7% of centers continued to recognize supine as the recommended position 6 months later. The percentage of centers that reported written sleep position policies increased from 18.8% to 44.4%. CONCLUSIONS: A targeted educational in-service for child care providers is effective in increasing awareness and knowledge, changing child care provider behavior, and promoting development of written sleep position policies. This change is sustained over at least a 6-month period. PMID- 14523181 TI - Where should infants sleep? A comparison of risk for suffocation of infants sleeping in cribs, adult beds, and other sleeping locations. AB - OBJECTIVES: To ascertain whether the number of sudden infant deaths as a result of suffocation in cribs, in adult beds, on sofas or chairs, and on other sleep surfaces was increasing whether attributable to increased reporting, diagnostic shift, or an actual increase in suffocation deaths and to compare the risk of reported accidental suffocation for infants on sleep surfaces designed for infants with the risk on adult beds. METHODS: We reviewed all accidental suffocation deaths among infants < or =11 months of age reported to the United States Consumer Product Safety Commission from 1980 through 1983 and 1995 through 1998. We compared infants' ages and other demographic data, the sleep location and surface used, and the reported mechanism or pattern of death. For 1995-1998, we used data on sleep location from an annual survey of randomly selected households of living infants younger than 8 months, collected as part of the National Infant Sleep Position Study at the National Institute of Child Health and Human Development, to calculate risk for death as a result of suffocation in cribs, in adult beds, and on sofas or chairs. METHODS: The number of reported suffocation deaths by location were compared between the 1980s and 1990s using logistic regression modeling to calculate odds ratios (OR), 95% confidence intervals (CI), and P values. Comparative risks for suffocation deaths on a given sleep surface for infants in the 1990s were examined by calculating rates of death per 100 000 exposed infants and comparing the 95% CI for overlap. RESULTS: From the 1980s, 513 cases of infant suffocation were considered; from the 1990s, 883 cases. The number of reported suffocation deaths in cribs fell from 192 to 107, the number of reported deaths in adult beds increased from 152 to 391, and the number of reported deaths on sofas or chairs increased from 33 to 110. Using cribs as the reference group and adjusting for potential confounders, the multivariate ORs showed that infant deaths in adult beds were 8.1 times more likely to be reported in the 1990s than in the 1980s (95% CI: 3.2-20.3), and infant deaths on sofas and chairs were 17.2 times more likely to be reported in the 1990s than in the 1980s (95% CI: 5.0-59.3). The sleep location of a subset of cases from the 1990s, 348 infants younger than 8 months at death, was compared with the sleep location of 4220 living infants younger than 8 months. The risk of suffocation was approximately 40 times higher for infants in adult beds compared with those in cribs. The increase in risk remained high even when overlying deaths were discounted (32 times higher) or the estimate of rates of bedsharing among living infants doubled (20 times higher). CONCLUSIONS: Reported deaths of infants who suffocated on sleep surfaces other than those designed for infants are increasing. The most conservative estimate showed that the risk of suffocation increased by 20-fold when infants were placed to sleep in adult beds rather than in cribs. The public should be clearly informed of the attendant risks. PMID- 14523182 TI - Are minority children under- or overrepresented in pediatric research? AB - OBJECTIVES: There is extensive documentation that minority adults are underrepresented in medical research, but there are scant data regarding minority children and their parents. METHODS: All full-length articles published in the paper edition of 3 general pediatric journals between July 1999 and June 2000 were collected and reviewed. Articles were excluded when they did not include at least 1 US researcher, all subjects at US institutions, parents or children as subjects, some prospective data collection, or between 8 and 10 000 subjects. We recorded the number and race/ethnicity (R/E) of all subjects, the type of research, and the type of data collected. Corresponding authors were surveyed to clarify R/E data. RESULTS: A total of 192 studies qualified. R/E data were reported in 114 (59%) studies, and survey data provided additional or new information in 25 studies resulting in R/E data in 128 (67%) articles accounting for 75% of the subjects. R/E was described by >10 different labels. There was an overrepresentation of black subjects and an underrepresentation of white and Hispanic subjects compared with the census data. When compared with research participation of child subjects, generally, black children were overrepresented and Hispanic children were underrepresented in clinical trials, and both were underrepresented in therapeutic research. Black and Hispanic children were overrepresented in potentially stigmatizing research. CONCLUSIONS: Overall, we found an overrepresentation of black subjects and an underrepresentation of white and Hispanic subjects with significant variations depending on the type of research. PMID- 14523183 TI - Plasma concentrations of aminoterminal pro atrial natriuretic peptide and aminoterminal pro brain natriuretic peptide in healthy neonates: marked and rapid increase after birth. AB - OBJECTIVE: The purpose of the study was to examine the possible age dependency of plasma N-terminal pro atrial natriuretic peptide (N-ANP) and N-terminal pro brain natriuretic peptide (N-BNP) levels in healthy term neonates to establish normal ranges for the neonatal period. METHODS: N-ANP and N-BNP plasma concentrations were measured in peripheral venous (n = 116) and umbilical cord blood (n = 37) in 153 healthy term neonates (mean: 5.1; range: 0-30 days) using an enzyme immunoassay. The neonates were classified into 8 groups according to their age (day of delivery and 1, 2, 3, 4, 5-7, 8-14, and 14-30 days of age). RESULTS: The plasma N-ANP and N-BNP concentration were the highest at the first day of age (96 700; 6912-436 000 and 641; 254-1272 fmol/mL) and were found significantly higher compared with the day of delivery (5680; 1005-16 900 and 221; 58-478 fmol/mL; P < 0,0001). After this marked increase, N-ANP and N-BNP levels decreased steadily and became stable at the fifth (5232; 2691-7353 fmol/mL) and third (246; 110-430 fmol/mL) day of life, respectively. CONCLUSIONS: The N-ANP and N-BNP plasma concentrations in healthy neonates showed a marked increase during the first days of age, suggesting that ANP and BNP have physiologic roles in the perinatal circulatory change from fetus to neonate. PMID- 14523184 TI - Relation between dieting and weight change among preadolescents and adolescents. AB - OBJECTIVE: To assess whether dieting to control weight was associated with weight change among children and adolescents. METHODS: A prospective study was conducted of 8203 girls and 6769 boys who were 9 to 14 years of age in 1996, were in an ongoing cohort study, and completed at least 2 annual questionnaires between 1996 and 1999. Dieting to control weight, binge eating, and dietary intake were assessed annually from 1996 through 1998 with instruments designed specifically for children and adolescents. The outcome measure was age- and sex-specific z score of body mass index (BMI). RESULTS: In 1996, 25.0% of the girls and 13.8% of the boys were infrequent dieters and 4.5% of the girls and 2.2% of the boys were frequent dieters. Among the girls, the percentage of dieters increased over the following 2 years. Binge eating was more common among the girls, but in both sexes, it was associated with dieting to control weight (girls: infrequent dieters, odds ratio [OR]: 5.10; frequent dieters, OR: 12.4; boys: infrequent dieters, OR: 3.49; frequent dieters, OR: 7.30). During 3 years of follow-up, dieters gained more weight than nondieters. Among the girls, frequency of dieting was positively associated with increases in age- and sex-specific z scores of BMI (beta = 0.05 and beta = 0.04 for frequent and infrequent dieters vs nondieters). Among the boys, both frequent and infrequent dieters gained 0.07 z scores of BMI more than nondieters. In addition, boys who engaged in binge eating gained significantly more weight than nondieters. CONCLUSIONS: Although medically supervised weight control may be beneficial for overweight youths, our data suggest that for many adolescents, dieting to control weight is not only ineffective, it may actually promote weight gain. PMID- 14523185 TI - Simple car seat insert to prevent upper airway narrowing in preterm infants: a pilot study. AB - OBJECTIVES: To test prospectively the hypothesis that an infant car seat modification to allow the infant's head to rest in a neutral position on the trunk would prevent narrowing of the upper airway and thus reduce oxygen desaturation in preterm infants who are restrained in car seats. METHODS: Seventeen preterm infants who were approved for discharge were evaluated in a car seat for newborns, with and without a foam insert that provided a slot for the back of the infants' head. Respiration timed inspiratory radiographs for assessment of upper airway dimensions were taken during quiet sleep in each position. Infants were monitored in each position for 30 minutes with continuous polygraphic recording of respiratory, cardiac, and nasal airflow activity and pulse oximetry. RESULTS: Placement of the insert in the car seat was associated with a larger upper airway space (mean +/- standard deviation, 5.2 +/- 1.3 vs 3.6 +/- 1.4 mm). This radiologic improvement was associated with a significant reduction in the frequency of episodes of oxygen desaturation to <85% (1.5 +/- 2.1 vs 3.5 +/- 3.5 episodes/infant), of bradycardia <90 bpm (0.1 +/- 0.3 vs 1 +/- 1.7), and of arousal (median [25th, 75th], 2.5 [1.3, 4.0] vs 5.0 [4.0, 7.0]). CONCLUSIONS: The cause of oxygen desaturation in preterm infants who are restrained in car seats is multifactorial. The present data strongly support the hypothesis that flexion of the head on the body is a significant contributor to these episodes and that the mechanism is posterocephalic displacement of the mandible, leading to narrowing of the upper airway. Critically, this pilot study demonstrates that the frequency of episodes of desaturation in a standard newborn car seat can be substantially reduced by placement of a simple foam insert that allows the infant to maintain the head in a neutral position on the trunk during sleep. PMID- 14523186 TI - Reducing acute adverse outcomes in youths with type 1 diabetes: a randomized, controlled trial. AB - OBJECTIVE: Both acute and chronic complications of diabetes account for a disproportionate percentage of US health care expenditures. Despite improvements in diabetes care, the incidence of adverse events in children with type 1 diabetes remains high, particularly for youths with poor glycemic control. Cost effective intervention programs designed to reduce complications are needed. This study evaluated a low-intensity, nonmedical intervention using a case manager (called a "Care Ambassador"), with and without the supplementation of psychoeducational modules, designed to monitor and encourage routine diabetes care visits to reduce short-term adverse outcomes and improve glycemic control in youths with type 1 diabetes. METHODS: We performed a 2-year prospective, randomized clinical trial in 299 youths with type 1 diabetes, aged 7 to 16 years, comparing 3 treatment programs (Care Ambassador [CA], Care Ambassador plus psychoeducational modules [CA+], and standard multidisciplinary diabetes care [SC]). The study was conducted in a large metropolitan US city from April 1997 through April 2000. Number of medical visits, frequency of hypoglycemic events, hospital/emergency department (ED) utilization, and glycosylated hemoglobin A1c were assessed during follow-up. RESULTS: During the 2-year study period, both the CA and CA+ groups had significantly more routine visits (mean [standard deviation]: 7.3 [2.06] and 7.5 [2.02], respectively) compared with the SC group (5.4 [2.62]). The CA+ intervention group had significantly reduced rates of short term adverse outcomes compared with the other 2 groups; 25% fewer total hypoglycemic events, 60% fewer severe hypoglycemic events, and 40% fewer hospitalizations and ED visits. "High-risk" youths in the CA+ group (baseline glycosylated hemoglobin A1c > or =8.7%) were 3.4-fold (1.57-7.41) more likely to improve their glycemic control compared with those at high risk in the other 2 groups. CONCLUSIONS: For youths with type 1 diabetes, the CA and CA+ interventions increased visit frequency. Youths in the CA+ intervention had reduced rates of hypoglycemia and hospital/ED utilization with estimated annual cost savings of 80 000 dollars to 90 000 dollars. The CA+ intervention compared with the other 2 groups improved glycemic control in "high-risk" youths. Nonmedical case management incorporating psychoeducational modules seems to be a cost-effective approach to improving outcomes in youths with diabetes. PMID- 14523187 TI - Congenital hypothyroidism: influence of disease severity and L-thyroxine treatment on intellectual, motor, and school-associated outcomes in young adults. AB - OBJECTIVE: To describe intellectual, motor, and school-associated outcome in young adults with early treated congenital hypothyroidism (CH) and to study the association between long-term outcome and CH variables acting at different points in time during early development (CH severity and early L-thyroxine treatment levels [0-6 years]). METHODS: Neuropsychological tests were administered to all 49 subjects with CH identified during the first 3 years of the Norwegian neonatal screening program (1979-1981) at a mean age of 20 years and to 41 sibling control subjects (mean age: 21 years). RESULTS: The CH group attained significantly lower scores than control subjects on intellectual, motor, and school-associated tests (total IQ: 102.4 [standard deviation: 13] vs 111.4 [standard deviation: 13]). Twelve (24%) of the 49 CH subjects had not completed senior high school, in contrast to 6% of the control subjects. CH severity (pretreatment serum thyroxine [T4]) correlated primarily with motor tests, whereas early L-thyroxine treatment levels were related to verbal IQ and school-associated tests. In multiple regression analysis, initial L-thyroxine dose (beta = 0.32) and mean serum T4 level during the second year (beta = 0.48) predicted Verbal IQ, whereas mean serum T4 level during the second year (beta = 0.44) predicted Arithmetic. CONCLUSIONS: Long-term outcome revealed enduring cognitive and motor deficits in young adults with CH relative to control subjects. Verbal functions and Arithmetic were associated with L-thyroxine treatment variables, suggesting that more optimal treatment might be possible. Motor outcome was associated with CH severity, indicating a prenatal effect. PMID- 14523188 TI - Adolescent assault injury: risk and protective factors and locations of contact for intervention. AB - BACKGROUND: Violence is a large contributor to morbidity and mortality among adolescents. Most studies assessing markers for violent injury are cross sectional. To guide intervention, we conducted a case-control study to explore factors associated with assault injury and locations to reach at-risk adolescents. OBJECTIVE: 1) To assess risk and protective factors for adolescent assault injury compared with 2 control groups of youth with unintentional injuries and noninjury complaints presenting to the emergency department and 2) to assess locations of contact with assault-injured youth for prevention programs. METHODS: Face-to-face and phone interviews were conducted with systematic samples of youth aged 12 to 19 years presenting to the emergency department with assault injury, unintentional injury, and noninjury complaints. Youth with intentional injuries were matched to youth in the 2 control groups on age +/-1 year, gender, race, and residency. RESULTS: One hundred forty-seven 147 assault-injured youth completed interviews. One hundred thirty-three assault injured youth were matched to 133 unintentionally injured and 133 noninjured youth presenting to the emergency department. Compared with the 2 control groups, assault-injured youth were more likely to have had more fights in the past year (odds ratio [OR]: 3.91; 95% confidence interval [CI]: 2.02, 7.58; OR: 4.00; 95% CI: 2.23, 7.18) and fights requiring medical treatment (OR: 35.49; 95% CI: 8.71, 144.68; OR: 80.00; 95% CI: 11.13, 574.80). Eighty percent of assault-injured youth had been in 1 or more fights in the last 12 months compared with 55% and 46% in unintentional and noninjured controls, respectively. Assault-injured youth were more likely to have had previous weapon injuries (OR: 9.50; 95% CI: 3.39, 26.6; OR: 8.50; 95% CI: 3.02, 23.95) and have seen someone shot (OR: 2.00; 95% CI 1.12, 3.58; OR: 2.00; 95% CI: 1.12, 3.58). Eighty-six percent of assault-injured youth had a regular health care provider with 82% reporting a visit within the last year. There were no differences between cases and controls with regard to physician contact, extracurricular activity involvement, school or church attendance, police contact, weapon access or weapon-carrying, or witnessing nonweapon-related violence. CONCLUSIONS: Fighting was common among all groups. Assault-injured youth were more likely to have had previous weapon injuries and were high-risk for future injury. Past fights, past fight injuries, and seeing someone else shot were markers associated with assault injury. Health providers do have access to at-risk teens for clinical risk assessment and intervention. PMID- 14523189 TI - Constipation with acquired megarectum in children with autism. AB - OBJECTIVE: Recent evidence suggests that autistic children may have significant gastrointestinal symptoms. Although constipation occurs in 2% to 5% of healthy children, its clinical diagnosis is often difficult in children with behavioral disorders. We thus aimed to assess the prevalence of fecal loading in autistic children with gastrointestinal symptoms and to identify possible predictors of constipation. METHODS: We studied abdominal radiographs of 103 autistic children (87 boys) who were referred for gastroenterological assessment, in comparison with 29 control radiographs from children who were referred to the emergency department, most with abdominal pain. Radiographs were scored independently, in blinded manner, by 4 pediatric gastroenterologists and a radiologist. The severity of constipation was determined using a validated index. Details of stool habit, abdominal pain, dietary history, and laxative use were obtained from case notes. RESULTS: The incidence of constipation in the control subjects with abdominal pain was higher than reported for normal children. Despite this, moderate or severe constipation was more frequent in the autistic group than in the control subjects (36% vs 10%). Analysis of rectosigmoid loading showed more striking differences (54.4% of autistic children had moderate/severe loading or acquired megarectum compared with 24.1% of control subjects). Multivariate regression analysis showed consumption of milk to be the strongest predictor of constipation in the autistic group, whereas stool frequency, gluten consumption, soiling, and abdominal pain were not predictive of constipation. CONCLUSIONS: Constipation is a frequent finding in children with gastrointestinal symptoms and autism, particularly in the rectosigmoid colon, often with acquired megarectum. The absence of any correlation between the clinical history and the degree of fecal impaction in autistic children confirms the importance of an abdominal radiograph in the assessment of their degree of constipation. PMID- 14523190 TI - School-age outcomes in children who were extremely low birth weight from four international population-based cohorts. AB - OBJECTIVE: The aim of this study was to determine whether learning and school problems in extremely low birth weight (ELBW) and reference children differ between cohorts in different countries. METHODS: Participants were 4 international population-based cohorts of ELBW survivors who were 500 to 1000 g birth weight from New Jersey, central-west Ontario, Bavaria, and Holland (n = 532) and were followed longitudinally from birth. Psychometric data were collected independently and prospectively and included at least 1 measure of cognitive status and 1 measure of achievement administered to the children between the ages of 8 and 11 years. Adjustments were made for comparison of all measures based on reference norms within each country. Information on special educational assistance and grade repetition was obtained from the parents. RESULTS: The overall follow-up rate was 84% (range: 74%-90%; n = 436). The proportion of children who performed within the normal range (> or =85) were as follows: IQ between 44% and 62%; reading between 46% and 81%; arithmetic between 31% and 76%; and spelling between 39% and 65%. Children from New Jersey had the lowest rates of cognitive and achievement deficits, and Bavarian children did less well in achievement scores relative to their peers and other cohorts. Despite these differences, more than half of all cohorts required special educational assistance and/or repeated a grade. CONCLUSIONS: School difficulties were found to be a serious sequelae of ELBW in all 4 countries, an observation that has social and economic implications. PMID- 14523191 TI - Computed tomography and radiation risks: what pediatric health care providers should know. AB - Computed tomography (CT) is an extremely valuable diagnostic tool. Recent advances, particularly multidetector technology, have provided increased and more diverse applications. However, there is also the potential for inappropriate use and unnecessary radiation dose. Because some data indicate that low-dose radiation (such as that in CT) may have a significant risk of cancer, especially in young children, it is important to limit CT radiation by following the ALARA (as low as reasonably achievable) principle. There is a variety of strategies to limit radiation dose, including performing only necessary examinations, limiting the region of coverage, and adjusting individual CT settings based on indication, region imaged, and size of the child. The pediatric health care provider has a pivotal role in the performance of CT and may be the only individual who discusses these important CT radiation issues with the child and family. For this reason, this article will summarize the issues with CT patterns of use and radiation risk, and provide dose reduction strategies pertinent to pediatric health care providers. PMID- 14523192 TI - Standards for child and adolescent immunization practices. National Vaccine Advisory Committee. PMID- 14523193 TI - Children, computed tomography radiation dose, and the As Low As Reasonably Achievable (ALARA) concept. PMID- 14523195 TI - Pets and childhood asthma--how should the pediatrician respond to new information that pets may prevent asthma? PMID- 14523194 TI - Underwater birth: missing the evidence or missing the point? PMID- 14523196 TI - Improved outcomes associated with early resuscitation in septic shock: do we need to resuscitate the patient or the physician? PMID- 14523197 TI - Child and adolescent immunizations: new recommendations, new standards, new opportunities. PMID- 14523198 TI - Does iron-deficiency anemia affect child development? PMID- 14523199 TI - Bullous myringitis: a case-control study. AB - Prior studies have shown that bullous myringitis (BM) accounts for <10% of acute otitis media (AOM) cases, and that the distribution of viral and bacterial pathogens in BM is similar to that in AOM without BM, except for a relative increase in the proportion of Streptococcus pneumoniae in BM. We studied 518 cases of AOM in children aged 6 months to 12 years. Using tele-otoscopy to assist the diagnosis, we identified 41 cases (7.9%) with BM. Children who had AOM with BM were older than AOM patients without BM (median age: 4.3 years vs 18 months). We compared 41 cases of AOM with BM to 41 control cases of age-, race-, and gender-matched AOM patients without BM. When compared with this matched control group, children with BM had more severe symptoms at the time of diagnosis and were more likely to have bulging of the tympanic membrane in the quadrants that were not obscured by the bulla. Children with AOM and BM may require aggressive pain management. Although parents and clinicians may agree that a watchful waiting approach is appropriate for older children with mild AOM, children experiencing painful AOM with BM may not be successful candidates for a watchful waiting approach, because parents may resist postponement of antibiotic therapy in children who are more symptomatic. PMID- 14523201 TI - Increasing immunization coverage. American Academy of Pediatrics Committee on Community Health Services. American Academy of Pediatrics Committee on Practice and Ambulatory Medicine. AB - Despite many recent advances in vaccine delivery, the goal for universal immunization set in 1977 has not been reached. In 2001, only 77.2% of US toddlers 19 to 35 months of age had received their basic immunization series of 4 doses of diphtheria and tetanus toxoids and acellular pertussis (DTaP) vaccine, 3 doses of inactivated poliovirus vaccine, 1 dose of measles-mumps-rubella (MMR) vaccine, and 3 doses of Haemophilus influenzae type b (Hib) vaccine. Children who are members of a racial or ethnic minority, who are poor, or who live in inner-city or rural areas have lower immunization rates than do children in the general population. Additional challenges to vaccine delivery include the introduction of new childhood vaccines, ensuring a dependable supply of vaccines, bolstering public confidence in vaccine safety, and sufficient compensation for vaccine administration. Recent research has demonstrated specific and practical changes physicians can make to improve their practices' effectiveness in immunizing children, including the following: 1) sending parent reminders for upcoming visits and recall notices; 2) using prompts during all office visits to remind parents and staff about immunizations needed at that visit; 3) repeatedly measuring practice-wide immunization rates over time as part of a quality improvement effort; and 4) having in place standing orders for registered nurses, physician assistants, and medical assistants to identify opportunities to administer vaccines. Pediatricians should work individually and collectively at local and national levels to ensure that all children receive all childhood immunizations on time. Pediatricians also can proactively communicate with parents to ensure they understand the overall safety and efficacy of vaccines. PMID- 14523200 TI - Longstanding obliterative panarteritis in Kawasaki disease: lack of cyclosporin A effect. AB - Kawasaki disease is a childhood vasculitis of medium-sized vessels, affecting the coronary arteries in particular. We have treated a therapy-resistant child who met all diagnostic criteria for Kawasaki disease. After the boy was given intravenous immunoglobulins and salicylates, as well as several courses of pulsed methylprednisolone, disease recurred and coronary artery lesions became progressively detectable. Cyclosporin A was started and seemed clinically effective. In contrast to the positive effect on inflammatory parameters, ie, C reactive protein and white blood cell counts, a novel plasma marker for cytotoxicity (granzyme B) remained elevated. Coronary disease progressed to fatal obstruction and myocardial infarction. Echocardiography, electrocardiograms, and myocardial creatine phosphokinase did not predict impending death. At autopsy an obliterative panarteritis was observed resulting from massive fibrointimal proliferation, affecting the aorta and several large and medium-sized arteries. Immunophenotypic analysis of the inflammatory infiltrates in arteries revealed mainly granzyme-positive cytotoxic T cells and macrophages in the intima and media, as well as nodular aggregates of T cells, B cells, and plasma cells in the adventitia of affected arteries. These findings further endorse the role of specific cellular and humoral immunity in Kawasaki disease. Unremitting coronary arteritis and excessive smooth muscle hyperplasia resulted in coronary occlusion despite the use of cyclosporin A. PMID- 14523202 TI - Principles of child health care financing. American Academy of Pediatrics Committee on Child Health Financing. AB - Child health care financing must maximize access to quality, comprehensive pediatric and prenatal health care. This policy statement replaces the 1998 policy statement by the same title. Changes reflect recent state and federal legislation that affect child health care financing. The principles outlined in the statement will be used to evaluate the changing structure of child health care financing. PMID- 14523203 TI - Caffeine dosage in preemies questioned. PMID- 14523204 TI - To dye or not to dye: a randomized, clinical trial of a triple dye/alcohol regime versus dry cord care. PMID- 14523205 TI - Amplitude-integrated electroencephalogram and cerebral injury. PMID- 14523206 TI - The FDA warning against prolonged sedation with propofol in children remains warranted. PMID- 14523207 TI - Infants born to mothers with severe acute respiratory syndrome. AB - Severe acute respiratory syndrome (SARS) is a newly discovered infectious disease caused by a novel coronavirus. During the community outbreak in Hong Kong, 5 liveborn infants were born to pregnant women with SARS. A systematic search for perinatal transmission of the SARS-associated coronavirus, including serial reverse transcriptase-polymerase chain reaction assays, viral cultures, and paired serologic titers, failed to detect the virus in any of the infants. In addition, none of the infants developed clinical, radiologic, hematologic, or biochemical evidence suggestive of SARS. One preterm infant developed jejunal perforation and another developed necrotizing enterocolitis with ileal perforation shortly after birth. This case series is the first report to describe the clinical course of the first cohort of liveborn infants born to pregnant women with SARS. PMID- 14523208 TI - A young infant with severe acute respiratory syndrome. AB - Severe acute respiratory syndrome (SARS), a new contagious respiratory disease associated with a novel coronavirus, has spread worldwide and become a global health concern after its first outbreak in Guangdong Province of the People's Republic of China in November 2002. The clinical presentation and the radiologic, hematologic, biochemical, and microbiologic findings of a 56-day-old male infant with SARS are described. Some clinical and laboratory features are similar to those reported in adult and pediatric patients. However, this infant had a more severe clinical course as compared with the older children. This is the youngest patient with symptomatic SARS reported to date. PMID- 14523209 TI - Children hospitalized with severe acute respiratory syndrome-related illness in Toronto. AB - OBJECTIVE: An outbreak of severe acute respiratory syndrome (SARS) occurred in the greater Toronto area between February and June 2003. We describe the clinical, laboratory, and epidemiologic features of children who were admitted to the Hospital for Sick Children, Toronto, with a presumptive diagnosis of suspect or probable SARS. METHODS: A prospective investigational study protocol was established for the management of children with a presumptive diagnosis of suspect or probable SARS. All were ultimately classified as having probable SARS, suspect SARS, or another cause on the basis of their epidemiologic exposure, clinical and radiologic features, and results of microbiologic investigations. RESULTS: Twenty-five children were included; 10 were classified as probable SARS and 5 were classified as suspect SARS, and in 10 another cause was identified. The exposure consisted of direct contact with at least 1 adult probable SARS case in 11 children, travel from a World Health Organization-designated affected area in Asia in 9 children, and presence in a Toronto area hospital in which secondary SARS spread had occurred in 5 children. The predominant clinical manifestations of probable cases were fever, cough, and rhinorrhea. With the exception of 1 teenager, none of the children developed respiratory distress or an oxygen requirement, and all made full recoveries. Mild focal alveolar infiltrates were the predominant chest radiograph abnormality. Lymphopenia; neutropenia; thrombocytopenia; and elevated alanine aminotransferase, aspartate aminotransferase, and creatine kinase were present in some cases. Nasopharyngeal swab specimens were negative for the SARS-associated coronavirus by an in-house reverse transcriptase-polymerase chain reaction in all 25 children. CONCLUSIONS: Our results indicate that SARS is a relatively mild and nonspecific respiratory illness in previously healthy young children. The presence of fever in conjunction with a SARS exposure history should prompt one to consider SARS as a possible diagnosis in children irrespective of the presence or absence of respiratory symptoms. Reverse-transcriptase polymerase chain reaction analysis of nasopharyngeal specimens seems to be of little utility for the diagnosis of SARS during the early symptomatic phase of this illness in young children. PMID- 14523210 TI - Parental knowledge, attitudes, and practices associated with not receiving hepatitis A vaccine in a demonstration project in Butte County, California. AB - OBJECTIVE: To determine hepatitis A vaccination coverage and factors associated with not receiving hepatitis A vaccine among children. METHODS: A random cluster sample survey was conducted of parents of children who attended kindergarten in Butte County, California, in 2000. Because of a history of recurrent epidemics, an aggressive hepatitis A vaccination program was ongoing during the time this study was conducted. Receipt of 1 or 2 doses of hepatitis A vaccine was studied. RESULTS: Of 896 surveys sent, 648 (72%) were completed. The vaccination coverage for at least 1 dose of hepatitis A vaccine was 398 (62%) and for 2 doses was 272 (42%). Factors associated with not receiving the vaccine included lack of provider recommendation (vs having recommendation; odds ratio [OR]: 7.8; 95% confidence interval [CI]: 4.9-12.2), not having heard of the vaccine (OR: 2.4; 95% CI: 1.2-4.9), and parent's not perceiving child is likely to get hepatitis A (vs perceiving child might get disease; OR: 2.1; CI: 1.6-2.9). CONCLUSIONS: Vaccination coverage among kindergartners did not reach high levels (ie, >90%), despite aggressive vaccination efforts in this community. Lack of provider recommendation and lack of parental awareness of hepatitis A vaccine were the 2 most significant factors associated with failure to receive vaccine. These findings will facilitate the development of vaccination strategies for communities in which hepatitis A vaccination is recommended. PMID- 14523211 TI - Umbilical cord care in premature infants: the effect of two different cord-care regimens (salicylic sugar powder vs chlorhexidine) on cord separation time and other outcomes. AB - OBJECTIVES: To evaluate the effect of 2 cord-care regimens (salicylic sugar powder vs chlorhexidine as a 4% detergent water solution) on cord separation time and other outcomes in preterm infants. METHODS: A prospective, randomized, controlled trial was conducted on 244 preterm newborns with a gestational age of <34 weeks and a birth weight of <2500 g. All preterm newborns were enrolled, regardless of their health condition. We excluded from the study infants whose conditions during the first hours of life required the catheterization of umbilical vessels. We also excluded from the general statistical analysis all newborns who had their programmed cord-care regimen changed because of the presence or the suspicion of omphalitis. On arrival at our neonatal intensive care unit or neonatal special care unit, infants were bathed thoroughly with a soap solution (Saugella, Guieu, Italy), and the umbilical cord (UC) was treated with 1 of the 2 antiseptic products chosen for the study. The stump was then folded and covered with common sterile, dry gauze and kept in place by an elastic net. Until cord detachment and at every diaper change, the cord stump was cleaned with sterile water and treated with the same product initially used for first time cord care. On the third day of life, we obtained an umbilical swab either from the base of the cord or from the umbilicus if the cord was already sloughed. Six weeks after birth, during hospitalization or during a follow-up visit if already discharged, all infants had a medical examination to check the umbilicus area. Cord separation time, changing of the programmed cord-care regimen, death, omphalitis, sepsis, cord bleeding, nurses' opinion on treatments efficacy, and UC colonization were measured. RESULTS: The cord separation time was significantly lower in infants who were treated with salicylic sugar powder (6 +/- 2 days) than in infants who were treated with chlorhexidine (9 +/- 2 days). The programmed cord-care regimen was changed in a significantly higher number of newborns in the chlorhexidine group (17) than in the salicylic sugar group (3). None of the newborns died, and we found only sporadic cases of sepsis (1 patient in each group) and omphalitis (1 patient in the chlorhexidine group). A significantly higher percentage of nurses were satisfied with the salicylic sugar powder treatment (98%) than with the chlorhexidine treatment (67%), notwithstanding a more frequent occurrence of slight cord scar bleeding in the salicylic sugar group (7.8%) than in the chlorhexidine group (4%). The rate of negative umbilical swabs was significantly higher in infants treated with salicylic sugar powder (73.1%) than with chlorhexidine (53%). CONCLUSIONS: In neonatal intensive care units and neonatal special care units of developed countries, salicylic sugar powder can be used effectively and safely for UC care of preterm infants. PMID- 14523212 TI - Epidemiology of terror-related versus non-terror-related traumatic injury in children. AB - OBJECTIVE: In the past 2 years hundreds of children in Israel have been injured in terrorist attacks. There is a paucity of data on the epidemiology of terror related trauma in the pediatric population and its effect on the health care system. The objective of this study was to review the accumulated Israeli experience with medical care to young victims of terrorism and to use the knowledge obtained to contribute to the preparedness of medical personnel for future events. METHODS: Data on all patients who were younger than 18 years and were hospitalized from October 1, 2000, to December 31, 2001, for injuries sustained in a terrorist attack were obtained from the Israel National Trauma Registry. The parameters evaluated were patient age and sex, diagnosis, type, mechanism and severity of injury, interhospital transfer, stay in intensive care unit, duration of hospitalization, and need for rehabilitation. Findings were compared with the general pediatric population hospitalized for non-terror related trauma within the same time period. RESULTS: During the study period, 138 children were hospitalized for a terror-related injury and 8363 for a non-terror related injury. The study group was significantly older (mean age: 12.3 years [standard deviation: 5.1] v 6.9 years [standard deviation: 5.3]) and sustained proportionately more penetrating injuries (54% [n = 74] vs 9% [n = 725]). Differences were also noted in the proportion of internal injuries to the torso (11% in the patients with terror-related trauma vs 4% in those with non-terror related injuries), open wounds to the head (13% vs 6%), and critical injuries (Injury Severity Score of 25+; 25% vs 3%). The study group showed greater use of intensive care unit facilities (33% vs 8% in the comparison group), longer median hospitalization time (5 days vs 2 days), and greater need for rehabilitative care (17% vs 1%). CONCLUSIONS: Terror-related injuries are more severe than non-terror related injuries and increase the demand for acute care in children. PMID- 14523213 TI - Fetal hemoglobin synthesis determined by gamma-mRNA/gamma-mRNA + beta-mRNA quantitation in infants at risk for sudden infant death syndrome being monitored at home for apnea. AB - OBJECTIVE: Fetal hemoglobin (HbF) levels in the hemolysates obtained from infants who died from sudden infant death syndrome (SIDS) are reported to be markedly increased compared with controls. This finding could have been explained by increased HbF synthesis caused by episodes of hypoxemia in the SIDS infants. A prospective study in a group of infants being monitored at home after an apparent life-threatening event (ALTE) and considered at increased risk for SIDS was conducted with an improved ribonuclease protection assay. The ribonuclease protection assay allowed for the quantitation of [gamma/(gamma+beta)]-globin mRNAs, which has a highly significant correlation with the levels of HbF synthesis. METHODS: Thirty-five infants who were admitted for an ALTE were included in the study. All infants were at home under surveillance with a cardiorespiratory monitor and followed in an apnea clinic with monthly appointments. Seventy-three blood samples were obtained between 38 and 61 weeks of postconceptional age. For control purposes, a similar group of 37 normal infants (99 samples) whose HbF synthesis was previously determined were included. RESULTS: Mean [gamma/(gamma+beta)]-globin mRNAs were increased in the ALTE group at 42 to 45 and 46 to 49 weeks of postconceptional age (mean: 55.2 +/- 17.4% and 33.9 +/- 14%) in comparison with HbF synthesis in controls (mean: 42.6 +/- 13.7% and 23.6 +/- 9.8%). CONCLUSIONS: The data obtained in this report from infants who were considered at risk for SIDS show that HbF synthesis is increased between 42 and 49 weeks of postconceptional age. Determining HbF synthesis as described in this study may have value as a marker for episodes of hypoxemia for certain infants who are at risk for SIDS. PMID- 14523214 TI - Pediatric viral human immunodeficiency virus type 1 RNA levels, timing of infection, and disease progression in African HIV-1-infected children. AB - OBJECTIVE: To describe plasma human immunodeficiency virus type 1 (HIV-1) RNA levels in African HIV-1-infected children in relation to the timing of infection and disease progression. METHODS: A retrospective cohort study was conducted of 80 children who were born to HIV-1-positive mothers and clinically followed from birth to 18 months of age in the ANRS 049 Ditrame project, Abidjan, Cote d'Ivoire (West Africa). The diagnosis and timing of pediatric HIV-1 infection were determined prospectively according to HIV-1 DNA polymerase chain reaction results. A total of 364 HIV-1 RNA viral load (VL) measurements were assessed retrospectively. Kaplan-Meier analyses and proportional hazards models were used to evaluate the prognostic value of pediatric VL and covariates for HIV disease progression or death. RESULTS: Mean initial positive VL was significantly lower among children who were infected in utero (4.94 log10/mL, n = 12) than in children who were infected later (5.6-6.1 log10/mL, n = 68). In the first 6 months after diagnosis, HIV-1 RNA levels peaked (> or =6 log10/mL), regardless of timing of infection. Then, a slow decline (overall slope, -0.076 log10 copies/mL/mo) was observed until 18 months of age. A 1 log10 higher value of the pediatric peak VL (risk ratio [RR]: 1.85; 95% confidence interval [CI]: 1.0-3.44) and of the maternal VL at delivery (RR: 1.90; CI: 1.16-3.12) were independently associated with an increased risk of rapid progression to acquired immune deficiency syndrome (AIDS) or death at 18 months of life (23 AIDS diagnoses and 31 deaths). Disease progression or death was more rapid for girls than for boys (RR: 2.26; CI: 1.39-4.96). CONCLUSIONS: In Africa, pediatric HIV-1 RNA levels are very close to those described in industrialized countries and seem to be predictive of AIDS stage or death, as in industrialized countries. With antiretroviral therapy becoming more widely available, the early identification and monitoring of pediatric HIV disease remains of paramount importance in Africa. PMID- 14523215 TI - A prospective study of bacillus Calmette-Guerin scar formation and tuberculin skin test reactivity in infants in Lima, Peru. AB - OBJECTIVES: To determine the sensitivity of the bacillus Calmette-Guerin (BCG) scar as an indicator of previous vaccination and to ascertain the tuberculin skin test (TST) response in infancy after vaccination in a community from an area hyperendemic for tuberculosis (TB). METHODS: In a birth cohort of healthy term infants from Lima, Peru, a single dose of BCG vaccine was administered within the first month of life. Scar formation was assessed biweekly during the first 6 months and again at 3 years after vaccination. TST response was evaluated 6 months after vaccination. RESULTS: Six months after vaccination, 99% (68) of the newborns exhibited a BCG scar (>2 mm). Scar size did not differ by sex, birth weight, age at vaccination, or nutritional status in the first 2 months. Eighty percent of the participants were found 3 years after vaccination, and all of them had a BCG scar. Mean TST reaction size 6 months after vaccination was 2.9 +/- 0.3 mm. No association was found between sex or age at BCG vaccination and TST size. Only 3 children had a TST >10 mm, and the 3 had a TB contact at home. CONCLUSIONS: The BCG scar was a sensitive indicator of vaccination status up to 3 years after the administration of the vaccine in the first month of life. Although nearly a quarter of the children had a TST response >5 mm 6 months after vaccination, TST reactions >10 mm did not occur in the absence of exposure to a person with tuberculosis. A cutoff of 10 mm should be used for disease control purposes in people who are born in countries where TB is endemic. PMID- 14523216 TI - Reduced risk of neonatal respiratory infections among breastfed girls but not boys. AB - OBJECTIVE: The effect of breastfeeding on community-acquired neonatal infections has not been well studied, although the neonatal period is one of special vulnerability to infectious pathogens. Respiratory tract infections are the neonatal infection most commonly diagnosed after nursery discharge. We therefore chose respiratory tract infections diagnosed after nursery discharge as representative of neonatal community-acquired infection and studied the impact of breastfeeding on this neonatal infection syndrome. METHODS: An unmatched nested case-control study was performed within a previously defined study cohort of 13 224 mother-infant pairs delivering between October 1, 1990, and March 31, 1998. Infants who were delivered at < 37 weeks' gestation were excluded. Neonatal respiratory tract infections were defined using modified National Nosocomial Infections Surveillance System criteria and were included in the case series when diagnosed after nursery discharge and at age < or =30 days. Infant feeding status during the first month of life was ascertained using automated text search of electronic medical records and was categorized as exclusive breastfeeding, mixed feeding, or exclusive formula feeding. RESULTS: A total of 241 neonatal respiratory tract infections were found, and 1205 control subjects were selected. Compared with control subjects, case infants were more often born during the winter respiratory syncytial virus season (48% vs 33%), more likely to have a sibling present (70% vs 54%), and more likely to be a member of a socioeconomically at-risk family (24% vs 18%). Case patients were less likely to be exclusively breastfed (38% vs 44%) and equally likely to be exposed to mixed feeding (35% vs 34%) relative to control subjects. When compared with formula feeding only, the odds ratio (OR) of exclusive breastfeeding was 0.70 (95% confidence interval [CI]: 0.49-0.99) and that of mixed feeding was 0.83 (95% CI: 0.58-1.2). However, when stratified by infant sex, the inverse association between breastfeeding and risk of neonatal respiratory tract infection was confined to neonatal girls, for whom the unadjusted ORs associated with breastfeeding only and mixed feeding were 0.5 (95% CI: 0.29-0.78) and 0.6 (95% CI: 0.35-0.93), respectively. There was no meaningful association between breastfeeding and risk of neonatal respiratory tract infection among neonatal boys, for whom the unadjusted ORs associated with breastfeeding only and mixed feeding were 1.1 (95% CI: 0.63-1.8) and 1.3 (95% CI: 0.74-2.1), respectively. After adjustment for year of birth, season of birth, siblings, and socioeconomic status, both exclusive breastfeeding and mixed feeding remained protective among girls, with ORs of 0.5 (0.29-0.78) and 0.6 (0.34-0.93), respectively. The corresponding ORs for boys were 1.1 (0.64-2.0) and 1.4 (0.78-2.4). CONCLUSIONS: Breastfeeding was inversely associated with reduced risk of neonatal respiratory tract infections in girls but not in boys. Breastfeeding may confer protection against some community-acquired infections as early as the first month of life. PMID- 14523218 TI - Determinants of nonsynostotic plagiocephaly: a case-control study. AB - OBJECTIVE: There has been a large increase in reported cases of nonsynostotic plagiocephaly in infants since the adoption of supine sleeping recommendations to prevent sudden infant death syndrome. The objective of this study was to identify and quantify the determinants of nonsynostotic plagiocephaly in infants. METHODS: One hundred infants who received a diagnosis of having nonsynostotic plagiocephaly were recruited as case patients and compared with 94 control subjects who were selected from a citywide database of infants. The infants all were aged between 2 and 12 months. Information concerning sociodemographic variables, obstetric factors, infant factors, and infant care practices was obtained by parental interview. RESULTS: Case patients were significantly more likely to be male (adjusted odds ratio [aOR]: 2.51; 95% confidence interval [CI]: 1.23-5.16), to be a firstborn (aOR: 2.94; 95% CI: 1.46-5.96), and to have been premature (aOR: 3.26; 95% CI: 1.02-10.47). In the first 6 weeks, they were more likely to have been sleeping in the supine position (aOR: 7.02; 95% CI: 2.98 16.53), not to have had the head position varied when put down to sleep (aOR: 7.11; 95% CI: 2.75-18.37), and to have had <5 minutes a day of tummy time (OR: 2.26; 95% CI: 1.03-5.00). Mothers of case patients were more likely to perceive their infants as less active (aOR: 3.23; 95% CI: 1.38-7.56), to have a developmental delay (aOR: 3.32; 95% CI: 1.01-10.85), and to have had a definite preferred head orientation at 6 weeks (aOR: 37.46; 95% CI: 8.44-166.32). Case mothers were more likely to have no or low educational qualifications (aOR: 5.61; 95% CI: 2.02-15.56), although they were more likely to have attended antenatal classes (aOR: 6.61; 95% CI: 1.59-27.47). CONCLUSIONS: Early identification of a preferred head orientation, which may indicate the presence of neck muscle dysfunction, may help prevent the development or further development of nonsynostotic plagiocephaly in infants. Plagiocephaly might also be prevented by varying the head position when putting the very young infant down to sleep and by giving supervised tummy time when awake. PMID- 14523217 TI - Geographic disparities in children's mental health care. AB - OBJECTIVE: It is widely believed that only a minority of vulnerable children and adolescents receive any mental health services. Although health care disparities associated with sociodemographic characteristics are well known, almost no information exists about another potentially important source of disparity for children: How does state of residence affect mental health service use? METHODS: Observational analysis was conducted using the 1997 and 1999 waves of the National Survey of America's Families (N = 45 247 children aged 6-17), a population survey fielded in 13 states and a smaller geographically dispersed sample. We studied 4 dependent variables: 1) use of any mental health services and number of visits among users; 2) need for mental health care, based on 6 items from the Child Behavior Checklist; 3) unmet need (no services among children with identified need); and 4) need among users of mental health services. RESULTS: Use of any mental health care differs >2-fold across states, ranging from 5% in California and Texas to >10% in Colorado and Massachusetts. The variation across states in service use and unmet need exceeds the differences across racial/ethnic groups or family income. For example, the odds ratio of unmet need in California versus Massachusetts is 3.04, compared with 2.33 between Hispanic and white children. Differences in population characteristics across states do not explain much of the observed geographic variation in mental health related outcomes for children. Perhaps the most disconcerting finding is that the differences in use are not paralleled by differences in need. Overall, there is no apparent relationship between levels of need and use of services across states. As a general rule, states with high rates of services do not have low levels of need or vice versa, although that situation exists. Alabama and Texas, for example, have higher rates of need and lower rates of use than the nation as a whole, whereas Washington state displays the opposite pattern. Even with the similar levels of need and service use, states differ in the effectiveness of their delivery system. Alabama and Mississippi have high rates of need and low levels of use, but rates of unmet need are not significantly higher in those 2 states than in the nation, whereas California, Florida, and Texas have the highest rates of unmet need. In California and Texas, children from high-income families are more likely to receive some mental health services than children from low-income families. In Alabama and Mississippi, as well as in the states with the lowest rates of unmet need (Colorado, Massachusetts, and Minnesota), the opposite is true: children from low-income families are much more likely to receive any mental health service than children from high-income families. CONCLUSIONS: Large differences from the national average across states in service use and unmet need are the rule, rather than the exception. National averages obscure large differences that can exceed the effects of race/ethnicity or income. The differences in the rates of use or unmet need are not driven by differences in the racial/ethnic or socioeconomic makeup across states but more likely are the result of differences in state policies and health care market characteristics. These state policies and health care market characteristics can interact with sociodemographic characteristics and affect how effectively resources are used. For states such as California and Texas that have the lowest rates of mental health service use, it may be less important to raise the rates of service use than to deliver them to the children with the highest need, predominantly black and Hispanic children and children in low-income families. PMID- 14523219 TI - Acute human immunodeficiency virus syndrome in an adolescent. AB - Acute human immunodeficiency virus (HIV) seroconversion illness is a difficult diagnosis to make because of its nonspecific and protean manifestations. We present such a case in an adolescent. A 15-year-old boy presented with a 5-day history of fever, sore throat, vomiting, and diarrhea. The patient also reported a nonproductive cough, coryza, and fatigue. The patient's only risk factor for HIV infection was a history of unprotected intercourse with 5 girls. Physical examination was significant for fever, exudative tonsillopharyngitis, shotty cervical lymphadenopathy, and palpable purpura on both feet. Laboratory studies demonstrated lymphopenia and mild thrombocytopenia. Hemoglobin, serum creatinine, and urinalysis were normal. The following day, the patient remained febrile. Physical examination revealed oral ulcerations, conjunctivitis, and erythematous papules on the thorax; the purpura was unchanged. Serologies for hepatitis B, syphilis, HIV, and Epstein-Barr virus were negative. Bacterial cultures of blood and stool and viral cultures of throat and conjunctiva showed no pathogens. Coagulation profile and liver enzymes were normal. Within 1 week, all symptoms had resolved. The platelet count normalized. Repeat HIV serology was positive, as was HIV DNA polymerase chain reaction. Subsequent HIV viral load was 350 000, and the CD4 lymphocyte count was 351/mm3. HIV is the seventh leading cause of death among people aged 15 to 24 in the United States, and up to half of all new infections occur in adolescents. Our patient presented with many of the typical signs and symptoms of acute HIV infection: fever, fatigue, rash, pharyngitis, lymphadenopathy, oral ulcers, emesis, and diarrhea. Other symptoms commonly reported include headache, myalgias, arthralgias, aseptic meningitis, peripheral neuropathy, thrush, weight loss, night sweats, and genital ulcers. Common seroconversion laboratory findings include leukopenia, thrombocytopenia, and elevated transaminases. The suspicion of acute HIV illness should prompt virologic and serologic analysis. Initial serology is usually negative. Diagnosis therefore depends on direct detection of the virus, by assay of viral load (HIV RNA), DNA polymerase chain reaction, or p24 antigen. Both false-positive and false-negative results for these tests have been reported, further complicating early diagnosis. Pediatricians should play an active role in identifying HIV infected patients. Our case, the first report of acute HIV illness in an adolescent, emphasizes that clinicians should consider acute HIV seroconversion in the appropriate setting. Recognition of acute HIV syndrome is especially important for improving prognosis and limiting transmission. It is imperative that we maintain a high index of suspicion as primary care physicians for adolescents who present with a viral syndrome and appropriate risk factors. PMID- 14523220 TI - Safety of live viral vaccines in patients with chromosome 22q11.2 deletion syndrome (DiGeorge syndrome/velocardiofacial syndrome). AB - The package inserts of live viral vaccines include immunodeficiency as a contraindication. Nevertheless, patients with mild forms of immunodeficiency may benefit from vaccination. No published guidelines exist for the administration of these vaccines specifically to patients with chromosome 22q11.2 deletion syndrome. This syndrome is also sometimes called DiGeorge syndrome and is associated with thymic hypoplasia and diminished T-cell numbers and has a wide spectrum of phenotypic features that include cardiac anomalies, dysmorphic facial features, and hypocalcemia. Patients generally exhibit a mild to moderate decrement in T-cell numbers with preservation of T-cell function. The aims of this study were to investigate the incidence of side effects after live viral vaccine administration in a population with chromosome 22q11.2 deletion syndrome. The high frequency of this syndrome in the population (1:3000 children) mandates a greater understanding of the risks and benefits related to live viral vaccine administration. A retrospective analysis of vaccine adverse events was performed. The data acquisition form evaluated the frequency of live vaccine administration and the consequences of both vaccination and withholding the vaccine. Flow cytometric enumeration of T cells was performed as part of an immunologic evaluation. Thirty-two of 59 responders were vaccinated with the varicella vaccine. Only 9% of patients reported adverse events. However, 63% of unvaccinated children developed chickenpox. Comparison of patients who tolerated the vaccine with those who reported adverse events showed no statistically significant differences in current age (7 vs 5.7 years), age at vaccination (3 vs 2.5 years), or T-cell subset counts: CD3 (1951 vs 2083 cells/ microL), CD4 (1283 vs 1463 cells/ microL), and CD8 (530 vs 502 cells/ microL). Fifty-two of 59 responders were vaccinated with measles-mumps-rubella (MMR). Twelve (23%) of 52 reported mild side effects, including fever, rash, and constitutional symptoms. No severe adverse reactions were reported. No patient reported natural disease with measles, mumps, or rubella. There were no statistically significant differences between the T-cell counts in the vaccinated group reporting side effects versus the vaccinated group without side effects (mean CD3 counts: 1928 vs 1736 cells/ microL; CD4 counts: 1250 vs 1127 cells/ microL; and CD8 counts: 528 vs 483 cells/ microL). In our study, patients with chromosome 22q11.2 deletion syndrome had a similar incidence of adverse effects with varicella and MMR vaccines compared with that reported in the general population. All side effects were mild. However, in patients who did not receive the varicella vaccine, an overwhelming 63% contracted the disease. Patients who were not vaccinated against MMR did not develop natural disease. The data suggest that this is a cohort of patients with 22q11.2 deletion syndrome who have tolerated live viral vaccinations without evidence of significant side effects. A prospective study could address whether there are T-cell thresholds below which vaccination is unsafe; however, the information that we present suggests that vaccinating children with chromosome 22q11.2 deletion with live viral vaccines does not carry a significantly higher risk of adverse reactions compared with the general population, provided that they have no evidence of severe immunocompromise. PMID- 14523221 TI - Prevention and treatment of type 2 diabetes mellitus in children, with special emphasis on American Indian and Alaska Native children. American Academy of Pediatrics Committee on Native American Child Health. AB - The emergence of type 2 diabetes mellitus in the American Indian/Alaska Native pediatric population presents a new challenge for pediatricians and other health care professionals. This chronic disease requires preventive efforts, early diagnosis, and collaborative care of the patient and family within the context of a medical home. PMID- 14523222 TI - Ceramide, stress, and a "LAG" in aging. AB - Recent studies have implicated the longevity assurance gene LAG1 in ceramide synthesis. In light of a role for ceramide in yeast and mammalian stress responses and mammalian cellular senescence, important connections are emerging between ceramide and organismal aging. In this Perspective, we examine the evidence for these connections in yeast, Drosophila, and mammals, and speculate on their implications. PMID- 14523223 TI - A prospective, clinical and radiological study of early psoriatic arthritis: an early synovitis clinic experience. AB - OBJECTIVE: To determine the clinical presentation and clinical and radiological outcome of early psoriatic arthritis (PsA) at 1 and 2 yr. METHODS: Patients with PsA were assessed at the St. Vincent's University Hospital Early Synovitis Clinic. Standardized clinical and laboratory assessment was performed at presentation and 1- and 2-yr follow-up. Radiographs of the hands and feet were evaluated in chronological order by two trained observers using the Sharp method modified to include the distal interphalangeal (DIP) joints. RESULTS: A total of 129 (12.7%) of 1018 patients were diagnosed with PsA [mean age at onset of arthritis was 40.4 yr (range 11-76); mean duration of disease was 9.9 months (range 0.3-48); 52 oligoarticular, 77 polyarticular]. Means and standard deviations of indices of disease activity at presentation were: 10-cm visual analogue scale = 4.8 +/- 2.7, HAQ score = 0.71 +/- 0.64, ACR functional class III/IV = 41 (35%), Ritchie Articular Index = 5.6 +/- 6, swollen joint count = 6.9 +/- 8, erythrocyte sedimentation rate = 24 +/- 26.4 mm/h, C-reactive protein = 27.6 +/- 58.5 mg/l. At presentation, 49 (38%) patients had peripheral enthesopathy, 13 patients (10%) had inflammatory spine pain and 50 (39%) patients had DIP involvement. A total of 119 had psoriasis at the time of presentation [plaque psoriasis in 112 (94%), mean age of psoriasis onset was 29.8 +/- 16.2 yr, nail dystrophy present in 78 patients (67%)]. At 1 yr of follow-up, 119 (92%) patients were reassessed and 70 (59%) were taking a disease-modifying anti rheumatic drug (DMARD). At 2 yr, 97 (75%) patients were reassessed and 54 (56%) were taking a DMARD. Despite considerable improvement in inflammation and function scores, only 31 (26%) patients were in remission at 1 yr with 20 (21%) in remission at 2 yr. There was a low rate of DMARD-free remission [14 (12%) at 1 yr and 11 (11%) at 2 yr]. Radiographs of hands and feet were obtained for 117 (91%) patients at presentation and 86 (67%) patients at a median follow-up of 24 months (range 11-56); 47% of patients had joint erosions in hands or feet at follow-up with a mean Sharp erosion score of 3 (0) +/- 5.2 (range 0-25) and a mean Sharp narrowing score of 3.2 (0) +/- 7.5 (range 0-48). CONCLUSION: This study confirms that PsA is a chronic, progressive disease in the majority of patients. Despite clinical improvement with current DMARD treatment, PsA results in radiological damage in up to 47% of patients at a median interval of 2 yr. PMID- 14523224 TI - Developing a programme of transitional care for adolescents with juvenile idiopathic arthritis: results of a postal survey. AB - OBJECTIVES: To explore the transitional needs of adolescents with juvenile idiopathic arthritis (JIA), as perceived by a range of professionals, and to examine how these needs may be addressed within a structured programme of transitional care. METHODS: Postal surveys (n = 1670) were distributed to key professionals employed in health, social support, education and vocation. RESULTS: Surveys were completed by 478 individuals. The majority of respondents (91%) were currently active in the care of adolescents with JIA. Planning for transitional care was perceived to be important for both adolescents and parents and to require multidisciplinary involvement. Respondents rated a wide range of resources to be important in supporting adolescents, including self-medication teaching packages and social skills training. A number of barriers to providing transitional care were identified, including inadequate resources, coordination and training. CONCLUSION: Transitional care in the context of JIA is perceived as necessary by a wide variety of professionals. PMID- 14523225 TI - Automated counting of white blood cells in synovial fluid. AB - OBJECTIVES: To evaluate the performance of automated leucocyte (white blood cell; WBC) counting by comparison with manual counting. METHODS: The number of WBC was determined in heparinized synovial fluid samples by the use of (i) a standard urine cytometer (Kova) and a microscope (reference method) and (ii) a haematology analyser (Sysmex XE-2100; WBC/BASO and DIFF channels). Imprecision within and between days was determined by replicate analysis of a low (level A; WBC approximately 0.560 x 10(9)/l) and a high (level B; WBC approximately 1.081 x 10(9)/l) dedicated synovial fluid control (Quantimetrix). RESULTS: The WBC count of the DIFF channel was highly correlated with the WBC count of the microscopic reference method (r = 0.99; WBC analyser = 0.870 x WBC reference method + 0.413). In contrast, no agreement existed between WBC counts generated by the WBC/BASO channel of the analyser and the reference method (r = 0.52; WBC analyser = 0.008 x WBC reference method + 0.079). Within-day imprecision (4-7%) and between-day imprecision (10%) of the haematology analyser were smaller than the within-day imprecision (12%) and the between-day imprecision (20-22%) of the manual reference method. For manual counting, inter-observer coefficients of variation were 35.9% (control level A) and 21.0% (control level B). CONCLUSIONS: The WBC count in synovial fluid can be reliably determined using the DIFF channel of the Sysmex XE-2100. Automated counting of WBC in synovial fluid offers more precise and faster results than manual counting. PMID- 14523226 TI - Serum MMP-1 and TIMP-1 levels are increased in patients with psoriatic arthritis and their siblings. AB - OBJECTIVE: To determine matrix metalloproteinase-1 (MMP-1) and tissue-inhibitor metalloproteinase-1 (TIMP-1) serum levels in patients with psoriatic arthritis (PsA) and to compare this with their siblings and local blood donor controls. PsA is an interesting condition in which to study metalloproteinases because there are variations in the level of destructiveness, including a significant proportion of cases without destructive change. This is unlike rheumatoid arthritis (RA) which is more uniformly destructive and where MMP-1/TIMP-1 levels are known to be elevated. METHODS: MMP-1 and TIMP-1 serum levels were determined by enzyme-linked immunosorbent assay (ELISA) in (a) index cases with PsA (subtype: RA n = 43, distal interphalangeal disease n = 2, oligoarticular n = 15, spondyloarthropathy n = 9, enthesitis n = 1), (b) siblings with PsA, (c) siblings with psoriasis (Ps), (d) unaffected siblings and (e) local controls. Patients with Ps were divided according to the onset of disease: type I disease, onset before age 40 yr and type II, onset after age 40 yr. RESULTS: MMP-1 and TIMP-1 levels were significantly increased in both the index cases and the group including all siblings compared with the controls (P < 0.0001). There was no statistical difference in MMP-1 or TIMP-1 levels between index cases and their siblings. There was no difference in serum MMP-1 level between the different subtypes (Moll and Wright) of PsA, but there was an increased level of serum TIMP 1 in patients with rheumatoid pattern (P = 0.05). In the index cases there were increased levels of TIMP-1 in type II onset psoriasis (P = 0.03) but no difference in MMP-1 levels. CONCLUSION: MMP-1 and TIMP-1 serum levels are elevated in PsA. This is greatest in RA pattern PsA. These levels were also elevated in unaffected siblings suggesting that genetic factors may be important. TIMP-1 levels were elevated in psoriasis alone, more so in late onset psoriasis, suggesting that the pathological processes of early and late onset psoriasis may be different. PMID- 14523227 TI - Do rheumatology cost-effectiveness analyses make sense? PMID- 14523228 TI - Thrombomodulin allosterically modulates the activity of the anticoagulant thrombin. AB - Exosite 1 of thrombin consists of a cluster of basic residues (Arg-35, Lys-36, Arg-67, Lys-70, Arg-73, Arg-75, and Arg-77 in chymotrypsinogen numbering) that play key roles in the function of thrombin. Structural data suggest that the side chain of Arg-35 projects toward the active site pocket of thrombin, but all other residues are poised to interact with thrombomodulin (TM). To study the role of these residues in TM-mediated protein C (PC) activation by thrombin, a charge reversal mutagenesis approach was used to replace these residues with a Glu in separate constructs. The catalytic properties of the mutants toward PC were analyzed in both the absence and presence of TM and Ca2+. It was discovered that, with the exception of the Arg-67 and Lys-70 mutants, all other mutants activated PC with similar maximum rate constants in the presence of a saturating concentration of TM and Ca2+, although their affinity for interaction with TM was markedly impaired. The catalytic properties of the Arg-35 mutant were changed so that PC activation by the mutant no longer required Ca2+ in the presence of TM, but, instead, it was accelerated by EDTA. Moreover, the activity of this mutant toward PC was improved approximately 25-fold independent of TM. These results suggest that Arg-35 is responsible for the Ca2+ dependence of PC activation by the thrombin-TM complex and that a function for TM in the activation complex is the allosteric alleviation of the inhibitory interaction of Arg-35 with the substrate. PMID- 14523229 TI - Drosophila Gr5a encodes a taste receptor tuned to trehalose. AB - Recent studies have suggested that Drosophila taste receptors are encoded by a family of G protein-coupled receptor genes comprising at least 56 members. One of these genes, Gr5a, has been shown by genetic analysis to be required by the fly for behavioral and sensory responses to a sugar, trehalose. Here, we show that Gr5a is expressed in neurons of taste sensilla located on the labellum and legs. Expression is observed in most if not all labellar sensilla and suggests that many taste neurons express more than one receptor. We demonstrate by heterologous expression in a Drosophila S2 cell line that Gr5a encodes a receptor tuned to trehalose. This is the first functional expression of an invertebrate taste receptor. PMID- 14523230 TI - The L box regulon: lysine sensing by leader RNAs of bacterial lysine biosynthesis genes. AB - Expression of amino acid biosynthesis genes in bacteria is often repressed when abundant supplies of the cognate amino acid are available. Repression of the Bacillus subtilis lysC gene by lysine was previously shown to occur at the level of premature termination of transcription. In this study we show that lysine directly promotes transcription termination during in vitro transcription with B. subtilis RNA polymerase and causes a structural shift in the lysC leader RNA. We find that B. subtilis lysC is a member of a large family of bacterial lysine biosynthesis genes that contain similar leader RNA elements. By analogy with related regulatory systems, we designate this leader RNA pattern the "L box." Genes in the L box family from Gram-negative bacteria appear to be regulated at the level of translation initiation rather than transcription termination. Mutations of B. subtilis lysC that disrupt conserved leader features result in loss of lysine repression in vivo and loss of lysine-dependent transcription termination in vitro. The identification of the L box pattern also provides an explanation for previously described mutations in both B. subtilis and Escherichia coli lysC that result in lysC overexpression and resistance to the lysine analog aminoethylcysteine. The L box regulatory system represents an example of gene regulation using an RNA element that directly senses the intracellular concentration of a small molecule. PMID- 14523231 TI - Mutations in bone morphogenetic protein receptor 1B cause brachydactyly type A2. AB - Brachydactyly (BD) type A2 is an autosomal dominant hand malformation characterized by shortening and lateral deviation of the index fingers and, to a variable degree, shortening and deviation of the first and second toes. We performed linkage analysis in two unrelated German families and mapped a locus for BD type A2 to 4q21-q25. This interval includes the gene bone morphogenetic protein receptor 1B (BMPR1B), a type I transmembrane serinethreonine kinase. In one family, we identified a T599 --> A mutation changing an isoleucine into a lysine residue (I200K) within the glycine/serine (GS) domain of BMPR1B, a region involved in phosphorylation of the receptor. In the other family we identified a C1456 --> T mutation leading to an arginine-to-tryptophan amino acid change (R486W) in a highly conserved region C-terminal of the BMPR1B kinase domain. An in vitro kinase assay showed that the I200K mutation is kinase-deficient, whereas the R486W mutation has normal kinase activity, indicating a different pathogenic mechanism. Functional analyses with a micromass culture system revealed a strong inhibition of chondrogenesis by both mutant receptors. Overexpression of mutant chBmpR1b in vivo in chick embryos by using a retroviral system resulted either in a BD phenotype with shortening and/or missing phalanges similar to the human phenotype or in severe hypoplasia of the entire limb. These findings imply that both mutations identified in human BMPR1B affect cartilage formation in a dominant-negative manner. PMID- 14523232 TI - Mechanism and energetics of green fluorescent protein chromophore synthesis revealed by trapped intermediate structures. AB - Green fluorescent protein has revolutionized cell labeling and molecular tagging, yet the driving force and mechanism for its spontaneous fluorophore synthesis are not established. Here we discover mutations that substantially slow the rate but not the yield of this posttranslational modification, determine structures of the trapped precyclization intermediate and oxidized postcyclization states, and identify unanticipated features critical to chromophore maturation. The protein architecture contains a dramatic approximately 80 degrees bend in the central helix, which focuses distortions at G67 to promote ring formation from amino acids S65, Y66, and G67. Significantly, these distortions eliminate potential helical hydrogen bonds that would otherwise have to be broken at an energetic cost during peptide cyclization and force the G67 nitrogen and S65 carbonyl oxygen atoms within van der Waals contact in preparation for covalent bond formation. Further, we determine that under aerobic, but not anaerobic, conditions the Gly-Gly-Gly chromophore sequence cyclizes and incorporates an oxygen atom. These results lead directly to a conjugation-trapping mechanism, in which a thermodynamically unfavorable cyclization reaction is coupled to an electronic conjugation trapping step, to drive chromophore maturation. Moreover, we propose primarily electrostatic roles for the R96 and E222 side chains in chromophore formation and suggest that the T62 carbonyl oxygen is the base that initiates the dehydration reaction. Our molecular mechanism provides the basis for understanding and eventually controlling chromophore creation. PMID- 14523233 TI - Regulation of nitric oxide consumption by hypoxic red blood cells. AB - The homeostasis of nitric oxide (NO) is attained through a balance between its production and consumption. Shifts in NO bioavailability have been linked to a variety of diseases. Although the regulation of NO production has been well documented, its consumption is largely thought to be unregulated. Here, we have demonstrated that under hypoxic conditions, NO accelerates its own consumption by increasing its entry into RBCs. When RBCs were exposed to NO (1:400 NO/heme ratio) under hypoxic conditions to form HbFe(II)NO, the consumption rate of NO increased significantly. This increase in NO consumption converted the bioactivity of serotonin from a vasodilator to a vasoconstrictor in isolated coronary arterioles. We identified HbFe(II)NO as a potential mediator of accelerated NO consumption. Accelerated NO consumption by HbFe(II)NO-bearing RBCs may contribute to hypoxic pulmonary vasoconstriction and the rebound effect seen on termination of NO inhalation therapy. Furthermore, accelerated NO consumption may exacerbate ischemia-mediated vasospasm and nitrate tolerance. Finally, this phenomenon may be an evolved mechanism to stabilize the vasculature in sepsis. PMID- 14523234 TI - Human uptake and incorporation of an immunogenic nonhuman dietary sialic acid. AB - Humans are genetically unable to produce the sialic acid N-glycolylneuraminic acid (Neu5Gc), because of a mutation that occurred after our last common ancestor with great apes. Although Neu5Gc is presumed absent from normal humans, small amounts have been claimed to exist in human tumors and fetal meconium. We have generated an antibody with high specificity and avidity for Neu5Gc. Fetal tissues, normal adult tissues, and breast carcinomas from humans showed reactivity to this antibody, primarily within secretory epithelia and blood vessels. The presence of small amounts of Neu5Gc was confirmed by MS. Absent any known alternate pathway for its synthesis, we reasoned that these small amounts of Neu5Gc might originate from exogenous sources. Indeed, human cells fed with Neu5Gc incorporated it into endogenous glycoproteins. When normal human volunteers ingested Neu5Gc, a portion was absorbed and eliminated in urine, and small quantities were incorporated into newly synthesized glycoproteins. Neu5Gc has never been reported in plants or microbes to our knowledge. We found that Neu5Gc is rare in poultry and fish, common in milk products, and enriched in red meats. Furthermore, normal humans have variable amounts of circulating IgA, IgM, and IgG antibodies against Neu5Gc, with the highest levels comparable to those of the previously known anti-alpha-galactose xenoreactive antibodies. This finding represents an instance wherein humans absorb and metabolically incorporate a nonhuman dietary component enriched in foods of mammalian origin, even while generating xenoreactive, and potentially autoreactive, antibodies against the same molecule. Potential implications for human diseases are briefly discussed. PMID- 14523235 TI - Modular organization of directionally tuned cells in the motor cortex: is there a short-range order? AB - We investigated the presence of short-range order (<600 microm) in the directional properties of neurons in the motor cortex of the monkey. For that purpose, we developed a quantitative method for the detection of functional cortical modules and used it to examine such potential modules formed by directionally tuned cells. In the functional domain, we labeled each cell by its preferred direction (PD) vector in 3D movement space; in the spatial domain, we used the position of the tip of the recording microelectrode as the cell's coordinate. The images produced by this method represented two orthogonal dimensions in the cortex; one was parallel ("horizontal") and the other perpendicular ("vertical") to the cortical layers. The distribution of directionally tuned cells in these dimensions was nonuniform and highly structured. Specifically, cells with similar PDs tended to segregate into vertically oriented minicolumns 50-100 microm wide and at least 500 microm high. Such minicolumns aggregated across the horizontal dimension in a secondary structure of higher order. In this structure, minicolumns with similar PDs were approximately 200 microm apart and were interleaved with minicolumns representing nearly orthogonal PDs; in addition, nonoverlapping columns representing nearly opposite PDs were approximately 350 microm apart. PMID- 14523236 TI - Identification of biomarkers for ovarian cancer using strong anion-exchange ProteinChips: potential use in diagnosis and prognosis. AB - One hundred eighty-four serum samples from patients with ovarian cancer (n = 109), patients with benign tumors (n = 19), and healthy donors (n = 56) were analyzed on strong anion-exchange surfaces using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry technology. Univariate and multivariate statistical analyses applied to protein-profiling data obtained from 140 training serum samples identified three biomarker protein panels. The first panel of five candidate protein biomarkers, termed the screening biomarker panel, effectively diagnosed benign and malignant ovarian neoplasia [95.7% sensitivity, 82.6% specificity, 89.2% accuracy, and receiver operating characteristic (ROC) area under the curve of 0.94]. The other two panels, consisting of five and four candidate protein biomarkers each, effectively distinguished between benign and malignant ovarian neoplasia and were therefore referred to as validation biomarker panel I (81.5% sensitivity, 94.9% specificity, 88.2% accuracy, and ROC = 0.94) and validation biomarker panel II (72.8% sensitivity, 94.9% specificity, 83.9% accuracy, and ROC = 0.90). The three ovarian cancer biomarker protein panels correctly diagnosed 41 of the 44 blinded test samples: 21 of 22 malignant ovarian neoplasias [10 of 11 early-stage ovarian cancer (I/II) and 11 of 11 advanced-stage ovarian cancer (III/IV)], 6 of 6 low malignant potential, 5 of the 6 benign tumors, and 9 of 10 normal patient samples. In conclusion, we have discovered three ovarian cancer biomarker protein panels that, when used together, effectively distinguished serum samples from healthy controls and patients with either benign or malignant ovarian neoplasia. PMID- 14523237 TI - Positive selection on protein-length in the evolution of a primate sperm ion channel. AB - Positive Darwinian selection on advantageous point substitutions has been demonstrated in many genes. We here provide empirical evidence, for the first time, that positive selection can also act on insertion/deletion (indel) substitutions in the evolution of a protein. CATSPER1 is a voltage-gated calcium channel found exclusively in the plasma membrane of the mammalian sperm tail and it is essential for sperm motility. We determined the DNA sequences of the first exon of the CATSPER1 gene from 15 primates, which encodes the intracellular N terminus region of approximately equal to 400 aa. These sequences exhibit an excessively high frequency of indels. However, all indels have lengths that are multiples of 3 nt (3n indels) and do not disrupt the ORF. The number of indel substitutions per site per year in CATSPER1 is five to eight times the corresponding rates calculated from two large-scale primate genomic comparisons, which represent the neutral rate of indel substitutions. Moreover, CATSPER1 indels are considerably longer than neutral indels. These observations strongly suggest that positive selection has been promoting the fixation of indel mutations in CATSPER1 exon 1. It has been shown in certain ion channels that the length of the N terminus region affects the rate of channel inactivation. This finding suggests that the selection detected may be related to the regulation of the CATSPER1 channel, which can affect sperm motility, an important determinant in sperm competition. PMID- 14523238 TI - Position-specific trapping of topoisomerase II by benzo[a]pyrene diol epoxide adducts: implications for interactions with intercalating anticancer agents. AB - DNA topoisomerase II (Top2) is the target of some of the most effective anticancer DNA intercalators. To determine the effect of intercalating ligands at defined positions relative to a known DNA cleavage site for human Top2alpha, we synthesized oligodeoxynucleotides containing single trans-opened benzo[a]pyrene 7,8-diol 9,10-epoxide (DE) deoxyadenosine (dA) adducts of known absolute configuration, placed at specific positions in a duplex sequence containing staggered Top2 cleavage sites on both strands. Because the orientations of the intercalated hydrocarbon are known from NMR solution structures of duplex oligonucleotides containing these dA adducts, a detailed analysis of the relationship between the position of intercalation and trapping of Top2 is possible. Our findings demonstrate that (i) Top2 cleavage complexes are trapped by intercalation of the hydrocarbon at either of the staggered cleavage sites or immediately adjacent to the base pairs flanking the cleavage sites within the stagger; (ii) both concerted and nonconcerted cleavage by both subunits of a Top2 homodimer were detected depending on the position of the benzo[a]pyrene DE dA adduct; and (iii) intercalation immediately outside of the staggered Top2 cleavage site, and to a lesser extent in the middle of the stagger, prevents Top2 from cleaving DNA at this site, consistent with the effect of some intercalators as suppressors of Top2-mediated DNA cleavage. These results identify specific binding sites for intercalators that result in trapping of Top2. Such poisoning of Top2 by bulky polycyclic aromatic hydrocarbon DE adducts constitutes a potential mechanism for their carcinogenic activity. PMID- 14523239 TI - Protein kinase C phosphorylation sensitizes but does not activate the capsaicin receptor transient receptor potential vanilloid 1 (TRPV1). AB - Protein kinase C (PKC) modulates the function of the capsaicin receptor transient receptor potential vanilloid 1 (TRPV1). This modulation manifests as increased current when the channel is activated by capsaicin. In addition, studies have suggested that phosphorylation by PKC might directly gate the channel, because PKC-activating phorbol esters induce TRPV1 currents in the absence of applied ligands. To test whether PKC both modulates and gates the TRPV1 function by direct phosphorylation, we used direct sequencing to determine the major sites of PKC phosphorylation on TRPV1 intracellular domains. We then tested the ability of the PKC-activating phorbol 12-myristate 13-acetate (PMA) to potentiate capsaicin induced currents and to directly gate TRPV1. We found that mutation of S800 to alanine significantly reduced the PMA-induced enhancement of capsaicin-evoked currents and the direct activation of TRPV1 by PMA. Mutation of S502 to alanine reduced PMA enhancement of capsaicin-evoked currents, but had no effect on direct activation of TRPV1 by PMA. Conversely, mutation of T704 to alanine had no effect on PMA enhancement of capsaicin-evoked currents but dramatically reduced direct activation of TRPV1 by PMA. These results, combined with pharmacological studies showing that inactive phorbol esters also weakly activate TRPV1, suggest that PKC mediated phosphorylation modulates TRPV1 but does not directly gate the channel. Rather, currents induced by phorbol esters result from the combination of a weak direct ligand-like activation of TRPV1 and the phosphorylation-induced enhancement of the TRPV1 function. Furthermore, modulation of the TRPV1 function by PKC appears to involve distinct phosphorylation sites depending on the mechanism of channel activation. PMID- 14523240 TI - DcpS can act in the 5'-3' mRNA decay pathway in addition to the 3'-5' pathway. AB - Eukaryotic mRNA degradation proceeds through two main pathways, both involving mRNA cap breakdown. In the 3'-5' mRNA decay pathway, mRNA body degradation generates free m7GpppN that is hydrolyzed by DcpS generating m7GMP. In the 5'-3' pathway, the recently identified human Dcp2 decapping enzyme cleaves the cap of deadenylated mRNAs to produce m7GDP and 5'-phosphorylated mRNA. We investigated mRNA decay in human cell extracts by using a new assay for decapping. We observed that 5'-phosphorylated intermediates resulting from decapping appear after incubation of a substrate RNA in human cell extracts, indicating the presence of an active 5'-3' mRNA decay pathway. Surprisingly, however, the cognate m7GDP product was not detected, whereas abundant amounts of m7GMP were generated. Additional experiments revealed that m7GDP is, unexpectedly, efficiently converted to m7GMP in extracts from various organisms. The factor necessary and sufficient for this reaction was identified as DcpS in both yeast and human. m7GMP is thus a general, pathway-independent, by-product of eukaryotic mRNA decay. m7GDP breakdown should prevent misincorporation of methylated nucleotides in nucleic acids and could generate a unique indicator allowing the cell to monitor mRNA decay. PMID- 14523242 TI - Transition-path sampling of beta-hairpin folding. AB - We examine the dynamical folding pathways of the C-terminal beta-hairpin of protein G-B1 in explicit solvent at room temperature by means of a transition path sampling algorithm. In agreement with previous free-energy calculations, the resulting path ensembles reveal a folding mechanism in which the hydrophobic residues collapse first followed by backbone hydrogen-bond formation, starting with the hydrogen bonds inside the hydrophobic core. In addition, the path ensembles contain information on the folding kinetics, including solvent motion. Using the recently developed transition interface sampling technique, we calculate the rate constant for unfolding of the protein fragment and find it to be in reasonable agreement with experiments. The results support the validation of using all-atom force fields to study protein folding. PMID- 14523241 TI - Subpallial origin of a population of projecting pioneer neurons during corticogenesis. AB - Pyramidal neurons of the mammalian cerebral cortex are generated in the ventricular zone of the pallium whereas the subpallium provides the cortex with inhibitory interneurons. The marginal zone contains a subpial stream of migratory interneurons and two different classes of transient neurons, the pioneer neurons provided with corticofugal axons, and the reelin-expressing Cajal-Retzius cells. We found in cultured slices that the medial ganglionic eminence provides the reelin-negative pioneer neurons of the marginal zone. Pioneer neurons sent long projection axons that went through the cortical plate and reached the subplate and the lateral border of the lateral ganglionic eminence. In the cultured slices, pioneer neurons were functionally mature: they displayed a voltage-gated sodium current, expressed functional alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) receptors, and showed gamma-aminobutyric acid type A (GABAA) postsynaptic events that were modulated by presynaptic AMPA receptors. Pioneer neurons expressed the adhesion molecules L1 and TAG-1; the latter has been reported to control tangential migrations to the neocortex [Denaxa, M., Chan, C.-H., Schachner, M., Parnavelas, J. & Karagogeos, D. (2001) Development (Cambridge, U.K.) 128, 4635-4644], and we show here that the pioneer neurons of the marginal zone are the cellular substrate of such a function. Finally, we show that, in early corticogenesis, reelin controls both the tangential migration of cortical interneurons toward the cortical plate and the tangential migration of pioneer neurons toward the marginal zone. PMID- 14523243 TI - The near attack conformation approach to the study of the chorismate to prephenate reaction. AB - Standard free energies (DeltaGN degree) for formation of near attack conformers, those ground state conformers that can convert directly to the transition state, were calculated for the Claisen rearrangement of chorismate to prephenate in six different environments: water, wild-type enzymes from Bacillus subtilis and Escherichia coli, their Arg90Cit and Glu52Ala mutants, and the 1F7 catalytic antibody. Values of the calculated DeltaGN degrees and the experimentally determined activation energies (DeltaG++) are linearly related with the slope of approximately equal to 1. This demonstrates that the relative rate of the chorismate --> prephenate reaction is overwhelmingly dependent on the efficiency of formation of near attack conformers in the ground state. PMID- 14523244 TI - Gleevec inhibits beta-amyloid production but not Notch cleavage. AB - Amyloid-beta (Abeta) peptides, consisting mainly of 40 and 42 aa (Abeta40 and Abeta42, respectively), are metabolites of the amyloid precursor protein and are believed to be major pathological determinants of Alzheimer's disease. The proteolytic cleavages that form the Abeta N and C termini are catalyzed by beta secretase and gamma-secretase, respectively. Here we demonstrate that gamma secretase generation of Abeta in an N2a cell-free system is ATP dependent. In addition, the Abl kinase inhibitor imatinib mesylate (Gleevec, or STI571), which targets the ATP-binding site of Abl and several other tyrosine kinases, potently reduces Abeta production in the N2a cell-free system and in intact N2a cells. Both STI571 and a related compound, inhibitor 2, also reduce Abeta production in rat primary neuronal cultures and in vivo in guinea pig brain. STI571 does not inhibit the gamma-secretase-catalyzed S3 cleavage of Notch-1. Furthermore, production of Abeta and its inhibition by STI571 were demonstrated to occur to similar extents in both Abl-/- and WT mouse fibroblasts, indicating that the effect of STI571 on Abeta production does not involve Abl kinase. The efficacy of STI571 in reducing Abeta without affecting Notch-1 cleavage may prove useful as a basis for developing novel therapies for Alzheimer's disease. PMID- 14523245 TI - In search of the molecular basis of heterosis. PMID- 14523246 TI - Dual regulated RNA transport pathways to the cortical region in developing rice endosperm. AB - Prolamine and glutelin RNAs are localized to two subdomains of the cortical endoplasmic reticulum (ER), the protein body ER and the cisternal ER, in developing rice seeds. The addition of nearly full-length prolamine sequences at the 3' untranslated region of a reporter RNA redirects its localization from the cisternal ER to the protein body ER. Deletion analysis of prolamine RNA sequences indicates the presence of two partially redundant cis elements required for protein body ER targeting. The addition of glutelin 3' untranslated region to protein body ER cis sequences, however, redirects RNA localization to the cisternal ER. These results indicate that there are at least two regulated RNA transport pathways as well as a constitutive pathway to the cortical ER. PMID- 14523247 TI - RPS4-mediated disease resistance requires the combined presence of RPS4 transcripts with full-length and truncated open reading frames. AB - Arabidopsis RPS4 belongs to the Toll/interleukin-1 receptor (TIR)-nucleotide binding site (NBS)-Leu-rich repeat (LRR) class of disease resistance (R) genes. Like other family members in different plant species, RPS4 produces alternative transcripts with truncated open reading frames. The dominant alternative RPS4 transcripts are generated by retention of intron 3 or introns 2 and 3, which contain in-frame stop codons and lie downstream of the NBS-encoding exon. We analyzed the biological significance of these alternative transcripts in disease resistance by removing introns 2 and 3, either individually or in combination, from a functional RPS4-Ler (Landsberg erecta) transgene. Removal of one or both introns abolished the function of the RPS4 transgene, whereas expression was not affected. In addition, a truncated RPS4-Ler transgene encoding the putative TIR and NBS domains was not sufficient to confer resistance, suggesting that the combined presence of regular and alternative RPS4 transcripts is necessary for function. Interestingly, we observed partial resistance in transgenic lines expressing both intron-deficient and truncated transgenes. This finding confirms the requirement for regular and alternative RPS4 transcripts and indicates that alternative transcripts function at the protein level rather than as regulatory RNAs. Together with published results on the tobacco N gene, our data suggest that the generation of alternative TIR-NBS-LRR R gene transcripts is of general biological significance across plant species. PMID- 14523248 TI - The Arabidopsis dynamin-like proteins ADL1C and ADL1E play a critical role in mitochondrial morphogenesis. AB - Dynamin-related proteins are high molecular weight GTP binding proteins and have been implicated in various biological processes. Here, we report the functional characterization of two dynamin homologs in Arabidopsis, Arabidopsis dynamin-like 1C (ADL1C) and Arabidopsis dynamin-like 1E (ADL1E). ADL1C and ADL1E show a high degree of amino acid sequence similarity with members of the dynamin family. However, both proteins lack the C-terminal Pro-rich domain and the pleckstrin homology domain. Expression of the dominant-negative mutant ADL1C[K48E] in protoplasts obtained from leaf cells caused abnormal mitochondrial elongation. Also, a T-DNA insertion mutation at the ADL1E gene caused abnormal mitochondrial elongation that was rescued by the transient expression of ADL1C and ADL1E in protoplasts. In immunohistochemistry and in vivo targeting experiments in Arabidopsis protoplasts, ADL1C and ADL1E appeared as numerous speckles and the two proteins colocalized. These speckles were partially colocalized with F1 ATPase-gamma:RFP, a mitochondrial marker, and ADL2b localized at the tip of mitochondria. These results suggest that ADL1C and ADL1E may play a critical role in mitochondrial fission in plant cells. PMID- 14523249 TI - Blue light-dependent in vivo and in vitro phosphorylation of Arabidopsis cryptochrome 1. AB - Cryptochromes are photolyase-like blue/UV-A light receptors that regulate various light responses in animals and plants. Arabidopsis cryptochrome 1 (cry1) is the major photoreceptor mediating blue light inhibition of hypocotyl elongation. The initial photochemistry underlying cryptochrome function and regulation remain poorly understood. We report here a study of the blue light-dependent phosphorylation of Arabidopsis cry1. Cry1 is detected primarily as unphosphorylated protein in etiolated seedlings, but it is phosphorylated in plants exposed to blue light. Cry1 phosphorylation increases in response to increased fluence of blue light, whereas the phosphorylated cry1 disappears rapidly when plants are transferred from light to dark. Light-dependent cry1 phosphorylation appears specific to blue light, because little cry1 phosphorylation is detected in seedlings treated with red light or far-red light, and it is largely independent from phytochrome actions, because no phytochrome mutants tested significantly affect cry1 phosphorylation. The Arabidopsis cry1 protein expressed and purified from insect cells is phosphorylated in vitro in a blue light-dependent manner, consistent with cry1 undergoing autophosphorylation. To determine whether cry1 phosphorylation is associated with its function or regulation, we isolated and characterized missense cry1 mutants that express full length CRY1 apoprotein. Mutant residues are found throughout the CRY1 coding sequence, but none of these inactive cry1 mutant proteins shows blue light induced phosphorylation. These results demonstrate that blue light-dependent cry1 phosphorylation is closely associated with the function or regulation of the photoreceptor and that the overall structure of cry1 is critical to its phosphorylation. PMID- 14523251 TI - Measurement: a beginner's guide. AB - This paper provides an introduction to measurement theory for psychometricians. The central concept in measurement theory is that of a continuous quantitative attribute and explaining what measurement is requires showing how this central concept leads on to those of ratio and real number and distinguishing measurements from measures. These distinctions made, the logic of quantification is described with particular emphasis upon the scientific task of quantification, as opposed to the instrumental task. The position presented is that measurement is the estimation of the magnitude of a quantitative attribute relative to a unit and that quantification is always contingent upon first attempting the scientific task of acquiring evidence that the relevant attribute is quantitative in structure. This position means that the definition of measurement usually given in psychology is incorrect and that psychologists' claims about being able to already measure psychological attributes must be seriously questioned. Just how the scientific task of investigating whether psychological attributes are quantitative may be undertaken in psychology is then considered and the corollary that psychological attributes may not actually be quantitative is raised. PMID- 14523252 TI - Rasch modeling and the measurement of social participation. AB - Social participation is the main outcome of physical rehabilitation programs. The aim of this study is to improve the measurement of social participation, using an instrument called the Assessment of Life Habits Scale and the Rasch model. The interval level measurement, the dimensionality and the generalizability of the item hierarchy were verified. The data from a large sample of people with spinal cord injury was analyzed and specific results were compared with expert opinions. The main properties of the instrument were satisfactory and the agreement with expert opinion was high. Principal component analysis showed multidimensionality. The item difficulty hierarchy obtained with spinal cord injury experts was different from the one obtained with traumatic brain injury experts, indicating a different difficulty level of items in relation to each population characteristics. We conclude that the instrument is appropriate for the measurement of social participation and suggest ways to improve the instrument. PMID- 14523253 TI - Measuring client satisfaction with public education III: group effects in client satisfaction. AB - A contracted research project to evaluate client satisfaction with public education (King and Bond, 2003; Bond and King, 2003) required that the satisfaction of certain groups of clients received particular attention. A number of target groups were specifically identified by the state education department as those requiring separate satisfaction analyses. ConQuest software (Wu, Adams and Wilson, 1997) provided convenient techniques for estimating group mean effects for identified sub-groups of the parent and student samples. This allowed for the analysis of directly comparable satisfaction estimates for groups such as these, as well as for school size, school type, school location and parents according to the year level of the child's school class. Rasch analyses of group mean effects revealed generally, that for students, differences in overall satisfaction levels between identified sub-samples and the whole student sample even if statistically significant, were, at most, substantively marginal. However, for identified groups of parents and caregivers, the results were not so equivocal: one group of indigenous Australians, and parents of children with disabilities showed marked positive group mean effects, while another group of indigenous Australians reported lower satisfaction levels than did the whole parent sample. Moreover, analyses based on groupings of schools according to their relative complexity, appeared to reiterate the finding mentioned en passant in a previous paper (King and Bond, 2003), that client satisfaction decreased as school size increased. PMID- 14523250 TI - The novel MYB protein EARLY-PHYTOCHROME-RESPONSIVE1 is a component of a slave circadian oscillator in Arabidopsis. AB - Using fluorescent differential display, we identified, from approximately 8000 displayed bands, a DNA fragment showing rapid induction in response to red light irradiation. This EARLY-PHYTOCHROME-RESPONSIVE1 gene (EPR1) encodes a novel nucleus-localized MYB protein harboring a single MYB domain that is highly similar to the circadian oscillator proteins CCA1 and LHY. EPR1 is regulated by both phytochrome A and phytochrome B, and the red-light induction of EPR1 is not inhibited by cycloheximide, demonstrating that EPR1 represents a primary phytochrome-responsive gene. Our results show that EPR1 overexpression results in enhanced far-red light-induced cotyledon opening and delayed flowering. In wild type Arabidopsis plants grown in continuous light, the EPR1 transcript exhibits circadian rhythmicity similar to that of CCA1 and LHY. Moreover, EPR1 suppresses its own expression, suggesting that this protein is part of a regulatory feedback loop. Constitutive expression of CCA1 and LHY results in the loss of EPR1 rhythmicity, whereas increased levels of EPR1 have no effect on the central oscillator. We propose that EPR1 is a component of a slave oscillator that contributes to the refinement of output pathways, ultimately mediating the correct oscillatory behavior of target genes. PMID- 14523254 TI - Toward a hierarchical goal theory model of school motivation. AB - Instead of concentrating on mastery and performance goal orientations, recent research on school motivation has suggested a multidimensional structure of achievement goal orientations. Students in Australian high schools (N =774) responded to 35 survey items on 10 goal orientation constructs (effort, task, sense of purpose, praise, competition, power, token, social concern, social dependence, and affiliation) and 14 items on general mastery, general performance, general social, and global motivation constructs. Confirmatory factor analysis results supported a hierarchical, multidimensional school motivation construct. The hierarchical, multidimensional model has provided a strong theoretical structure for further school motivation research. PMID- 14523255 TI - Examining reliability and validity of job analysis survey data. AB - Historically, job analysis has played a fundamental role for developing and validating licensure and certification examinations. Still, research on what constitutes reliable and valid job analysis data is lacking. This paper illustrates several ways to examine the reliability and validity of job analysis survey results. Generalizability theory and the many-facets Rasch model are applied to investigate consistency and generalizability in task importance measures, to suggest reliable sample size, and to justify the number and use of rating scales. By using random samples from job analysis data for two professions with divergent job activities, this study finds that a representative sample as small as 400 respondents produces reliable estimates of task importance to the same degree of generalizability as obtained from a larger sample of job analysis respondents. Analyses of rating scales suggest that the effectiveness of using different numbers and types of rating scales depends on the nature of a profession. PMID- 14523256 TI - Measuring coping at a university using a Rasch model. AB - Coping with academic difficulties at a university was based on six aspects: Motivation and Planning, Friends and Planning, Studying and Planning, Emotions, Spiritual Help and Coping by Doing Nothing. Stem-items for each aspect were conceptually ordered by difficulty. Each of the stem-items was answered from three perspectives, Good Coping Strategies, Actual Coping Strategies, and Stress Reduction Strategies. The three response categories were No, not on any occasion this semester; Yes, on 1 to 3 occasions this semester, and Yes, on 4 or more occasions this semester. The convenience sample was 337 students studying education at an Australian university and data were analysed with a Rasch measurement model. A scale was created in which the difficulties of the items were ordered from easy to hard and the student measures of Coping were ordered from low to high. Coping by Doing Nothing and Using Spiritual Help stem-items didn't fit the measurement model and were deleted. This left an effective item sample of 21 (7 stem-items times 3). The proportion of observed student variance considered true was 0.88. The results supported the theory behind the construct of Coping as using Motivation and Planning, Friends and Planning, Studying and Planning, and Emotions, in which Expected Good Coping Strategies are easier than Actual Coping Strategies which, in turn, are easier than Stress Reducing Coping Strategies. PMID- 14523257 TI - Detecting and measuring rater effects using many-facet Rasch measurement: part I. AB - The purpose of this two-part paper is to introduce researchers to the many-facet Rasch measurement (MFRM) approach for detecting and measuring rater effects. The researcher will learn how to use the Facets (Linacre, 2001) computer program to study five effects: leniency/severity, central tendency, randomness, halo, and differential leniency/severity. Part 1 of the paper provides critical background and context for studying MFRM. We present a catalog of rater effects, introducing effects that researchers have studied over the last three-quarters of a century in order to help readers gain a historical perspective on how those effects have been conceptualized. We define each effect and describe various ways the effect has been portrayed in the research literature. We then explain how researchers theorize that the effect impacts the quality of ratings, pinpoint various indices they have used to measure it, and describe various strategies that have been proposed to try to minimize its impact on the measurement of ratees. The second half of Part 1 provides conceptual and mathematical explanations of many-facet Rasch measurement, focusing on how researchers can use MFRM to study rater effects. First, we present the many-facet version of Andrich's (1978) rating scale model and identify questions about a rating operation that researchers can address using this model. We then introduce three hybrid MFRM models, explain the conceptual distinctions among them, describe how they differ from the rating scale model, and identify questions about a rating operation that researchers can address using these hybrid models. PMID- 14523258 TI - What is disease? In memory of Owsei Temkin. AB - This essay outlines a contextual approach to disease (and thus medicine) in society. The work of Owsei Temkin is retrospectively evaluated and shown to rest on an assumed (if often implicit) contextualism. The key components of historical contextualism are then articulated, including the historicity of disease, the reification of specific disease categories in terms of language and social practice, and finally, in contemporary society, the value placed on diagnosis, the bureaucratization of disease, and a logically consistent focus on boundary management and boundary disputes. It is a contextualism that demands a role for the biological as well as the cultural, for practice as well as pathological theory. PMID- 14523259 TI - Women and the practice and teaching of medicine in Bologna in the eighteenth and early nineteenth centuries. AB - When Bologna's political and intellectual elites had control over the university and associated institutions during the eighteenth century, a few women were allowed to participate in medical fields not typically associated with their gender. Laura Bassi could occasionally teach and debate on anatomy at the university, and the practical anatomist Anna Morandi Manzolini could make contributions to the research and teaching of anatomy. With the arrival of the French in Bologna in 1796, the control of the university and associated institutions passed from the local elite to the Ministry of Public Instruction in Milan. This article shows that a few women were awarded degrees in medicine, surgery, and pharmacy from the University of Bologna when the French were in control-however, these women, unlike Bassi and Morandi, were directed to fields deemed appropriate to their gender, such as the training of midwives, or apothecary practice within female institutions. PMID- 14523260 TI - The emergence of medical specialization in the nineteenth century. AB - This essay reexamines the nineteenth-century origins of medical specialization. It suggests that by the 1880s, specialization had become perceived as a necessity of medical science as a result of the realization of two preconditions: First, a new collective desire to expand medical knowledge prompted clinical researchers to specialize; only specialization, it was believed, permitted the rigorous observation of many cases. Second, administrative rationality suggested that one could best manage large populations through proper classification, gathering together individuals belonging to the same class and separating those belonging to different categories. Both of these conditions emerged first and most powerfully in early nineteenth-century Paris. They were, in contrast, uniquely underdeveloped in the fragmented medical community of London during this period. PMID- 14523261 TI - Why support a women's medical college? Philadelphia's early male medical pro feminists. AB - The male founders and early faculty of Philadelphia's Woman's Medical College were mostly abolitionist physicians, zealous moralists for whom medical feminism formed only one of the cherished causes they could "manfully" and righteously defend. Male faculty of the late nineteenth century comprised "self-made" men, mostly new specialists, for whom strict sexism probably seemed inconsistent with progressive medicine. For some of these physicians-obviously a small minority defending medical women and breaking the barriers of fraternity could be consistent with "manly" responsibility. The outcome of the collaboration of women and the dissident men physicians in nineteenth-century Philadelphia amounted to another seeming paradox: the majority of the male medical profession, both locally and nationally, tyrannically hindered women's entry into the profession, yet medicine opened its doors in advance of law and the clergy; and where this first occurred, such as in the community centered on Woman's Medical College, a novel gender rearrangement arose based on collaboration and friendship. PMID- 14523262 TI - Hay fever holiday: health, leisure, and place in Gilded-Age America. AB - By the 1880s hay fever (also called June Cold, Rose Cold, hay asthma, hay cold, or autumnal catarrh) had become the pride of America's leisure class. In mid August each year, thousands of sufferers fled to the White Mountains of New Hampshire, to the Adirondacks in upper New York State, to the shores of the Great Lakes, or to the Colorado plateau, hoping to escape the dreaded seasonal symptoms of watery eyes, flowing nose, sneezing fits, and attacks of asthma, which many regarded as the price of urban wealth and education. Through a focus on the White Mountains as America's most fashionable hay fever resort in the late nineteenth century, this essay explores the embodied local geography of hay fever as a disease. The sufferers found in the White Mountains physical relief, but also a place whose history affirmed their social identity and shaped their relationship to the natural environment. And, they, in turn, became active agents in shaping the geography of place: in the very material relationships of daily life, in the social contours of the region, and in the symbolic space that nature inhabited. In the consumption of nature for health and pleasure, this article suggests, lies an important, yet relatively unexplored, source for understanding changing perceptions of environment and place and the impact of health on the local and regional transformation of the North American landscape. PMID- 14523263 TI - "Unfit for human consumption": tuberculosis and the problem of infected meat in late Victorian Britain. AB - By the 1890s, questions about tuberculous meat in Britain served to transform the issue of infected meat from an ill-defined to a concrete threat. Veterinarians, building on European inoculation (or transmission) experiments, played a prominent part in constructing the debate, with medical officers of health following. With the emergence of bacteriology in the 1880s, a consensus emerged about the dangers of tuberculous meat: Robert Koch's identification of the tubercle bacillus in 1882, and the connection he saw between bovine tuberculosis and the disease in man, provided confirmation of the disease's danger to man. It was from this point that veterinary and public health interests diverged. Whereas a general agreement had been reached, the extent of the problem remained open to doubt. Confusion revolved around two issues: the localization of infection, and the question of cooking. The latter was thought to make tuberculous meat "safe," as attention shifted to the problem of milk; whereas the former frustrated efforts to combat the sale of meat showing signs of infection. PMID- 14523266 TI - The construction of emergent order, or, how to resist the temptation of hylozoism. AB - Hylozoism, the doctrine that nature is imbued by life even in the apparently inert and lifeless, has as one of its "appealing" features the ability to duck the perplexing issue of how life originated. By so doing, hylozoism has generated even more conundrums such as why rocks, if indeed they are actually animated, don't appear alive. Hylozoism is not so much a resolution as a way of avoiding the possibility of the emergence of the radically novel. But, hylozoism is not just a relic of the past since modern strains of its specific explanatory strategy can be detected in three examples from the study of complex systems, namely, May's and Feigenbaum's explanation of complexity and universality in "one humped" maps, Maturana's and Varela's idea of autopoeisis, and Goertzel's notion of self-generating systems. These three explanatory strategies are analyzed as to their hylozoist cast with the result that the first is found to be appropriate to the mathematical nature of the inquiry while the second two are found wanting in the same way that hylozoism in general proves unsatisfactory as an explanation. To remedy the problems associated with a hylozoist strategy, a constructional view of the emergence of new wholes is proposed including intimations as to how this constructional process might proceed. PMID- 14523267 TI - Common dynamic properties of biosignals during cognition: self-similarity and chaotic dynamics of both response times and EEG during movement imagery. AB - This study was undertaken to verify whether different output variables or biosignals, measured during performance of a cognitive task, manifest common dynamical properties. Nonlinear properties of both response times (RTs) and electroercephalograms (EEG) were tested. We asked subjects to generate mental images of actions following of auditorily presentation simple phrases suggesting the action. Analysis of RT series combined from many subjects and of EEG records from single subjects clearly manifested self-similarity and chaotic dynamics that provide insights into the self-organization of the brain/behavioral system. PMID- 14523268 TI - Heterochromatin and complexity: a theoretical approach. AB - Heterochromatin represents 30% of eukaryotic genome in Drosophila and 15% in humans. Despite extensive research spanning many decades, its evolutionary significance, as well as the forces that guarantee its maintenance, are still elusive. Many theoretical and experimental approaches have led researchers to propose several conceptual frameworks to elucidate the nature of this huge mysterious genetic material and its spreading in all eukaryotic genomes. "Junk DNA" as well as "selfish genetic material" are two examples of such attempts, but several lines of evidence suggest that such explanations are incomplete. In fact, if the selfish DNA hypothesis does not explain the mapping of genetic functions in heterochromatin, then the junk DNA hypothesis is incomplete in describing both emergence of genetic functions and their maintenance in the eukaryotic heterochromatin. Recent developments in the physics of complex systems and mathematical concepts such as fractals provide new conceptual clues to answer several basic questions concerning the emergence of heterochromatin in eukaryotic genomes, its evolutionary significance, the forces that guarantee its maintenance, and its peculiar behavior in the eukaryotic cell. The aim of this paper is to provide a new theoretical framework for the heterochromatin, considering such genetic material in physical terms as a complex adaptive system. We apply some computer calculations to demonstrate the nonlinearity of the flux of genetic information along the phylogenic tree. Fractal dimensions of representative heterochromatic sequences are provided. A theory is proposed in which heterochromatin is considered a system that evolves in a self-organized manner at the edge of cellular and environmental chaos. PMID- 14523271 TI - Detection of high frequencies of HIV-1 cross-subtype reactive CD8 T lymphocytes in the peripheral blood of HIV-1-infected Kenyans. AB - OBJECTIVES: To quantitate rapidly the frequency of HIV-1 subtype-specific and broadly HIV-1 cross-subtype-reactive CD8 T cells in the peripheral blood of HIV-1 infected individuals from a geographical region of multiple subtype endemicity. METHODS: Autologous B-lymphoblastoid cell lines infected with recombinant vaccinia-viruses expressing gag, env and nef gene products from HIV-1 subtypes A H were used as antigen-presenting cells to stimulate CD8 T cells from cryopreserved peripheral blood mononuclear cells. Cross-subtype and subtype specific CD8 cell responses were assessed by flow cytometry for the upregulation of IFN-gamma gene expression in specifically activated CD8 T cells. RESULTS: Strikingly high frequencies of circulating CD8 T cells (up to 11.3% of peripheral CD8 T cells) with specificity for HIV-1 were detectable using this methodology. Both subtype-specific and broadly cross-subtype-reactive CD8 T cells were detected as assessed by IFN-gamma production after stimulation. The pattern of cross-subtype reactivity appeared to be random when the results were assessed as a population, but analysis of the full-length sequence of the infecting virus for each individual showed some skewing of the CD8 cell response towards the infecting subtype. CONCLUSION: High frequencies of HIV-1 cross-subtype-reactive peripheral CD8 T cells can be detected in individuals from a multiple subtype endemic region, providing an incentive for vaccine advancement in such locations. The future assessment of the subtype specificity of cellular immune responses requires full-length sequencing of the infecting virus in conjunction with a comprehensive analysis of phenotypic and functional parameters. PMID- 14523272 TI - Antiviral dynamics and sex differences of zidovudine and lamivudine triphosphate concentrations in HIV-infected individuals. AB - OBJECTIVES: Nucleoside analog reverse transcriptase inhibitors (NRTI) are used in virtually all anti-HIV regimens. Clinical response depends on the intracellular formation of the pharmacologically active triphosphate moiety. Our objective was to quantify the pharmacological characteristics of zidovudine and lamivudine triphosphate in HIV-infected individuals. METHODS: Peripheral blood mononuclear cells were obtained at multiple planned intervals from antiretroviral-naive adults participating in a study of zidovudine, lamivudine and indinavir, and triphosphate levels were determined by immunoassay and high-performance liquid chromatography/mass spectrometry. Plasma HIV-RNA, CD4 cell counts, and plasma drug concentrations were collected over 18 months. Data were analysed using non parametric, regression and time-to-event methods. RESULTS: Thirty-three subjects were evaluated. The estimated half-lives of zidovudine and lamivudine triphosphate were 7 and 22 h, respectively. Triphosphate concentrations were elevated in individuals with low baseline CD4 cell counts. Triphosphate concentrations in women were higher than in men by 2.3 and 1.6-fold for zidovudine and lamivudine, respectively. Women reached an HIV-RNA level under 50 copies/ml twice as fast as men. Zidovudine triphosphate above 30 fmol/10(6) cells was independently predictive of the time to under 50 copies/ml. Lamivudine triphosphate above 7017 fmol/10(6) cells was independently predictive of a longer virological response. Indinavir concentrations were related to antiviral responses in univariate analyses. CONCLUSION: Zidovudine and lamivudine triphosphate concentration thresholds were independently associated with the antiviral activity of zidovudine, lamivudine, and indinavir. The significantly elevated triphosphate concentrations in women and individuals with low baseline CD4 cell counts, groups that historically experience high rates of serious NRTI toxicities, provide a hypothesis for the pathogenesis of these events. PMID- 14523273 TI - Correlates of HIV-1 shedding in cervicovaginal secretions and effects of antiretroviral therapies. AB - OBJECTIVES: To evaluate the determinants of HIV-1 RNA shedding in cervicovaginal secretions and the effects of antiretroviral therapy in a group of infected women. METHODS: A total of 122 women from whom paired peripheral blood and cervicovaginal lavage samples were available were enrolled in the study. HIV-1 RNA was quantified in the plasma and cell-free fraction of cervicovaginal lavages by the nucleic acid sequence-based amplification assay (lower limit of detection 80 copies/ml). RESULTS: Seventy-one per cent of the women had detectable viral load in the cervicovaginal lavage and this appeared to be correlated to plasma viral load and to the degree of immunodeficiency as expressed by the absolute number of CD4 cells. Antiretroviral-treated patients had a lower risk of shedding the virus in the genital tract, but this association was limited to patients treated with highly active antiretroviral therapy (HAART). However, in 25% of women with undetectable plasma viral load, a genital shedding of the virus was demonstrated. CONCLUSION: Plasma viral load may fail as a marker of infectivity of genital secretions. HAART treatment seems to be more efficacious in suppressing viral shedding at the genital level. The female genital tract represents a distinct compartment for HIV-1 replication/evolution. PMID- 14523274 TI - Differential effect of HIV protease inhibitors on adipogenesis: intracellular ritonavir is not sufficient to inhibit differentiation. AB - OBJECTIVES: Lipodystrophy is a major side effect of HIV protease inhibitor (PI) antiretroviral therapy. It has been shown that protease inhibitors interfere in vitro with adipocyte differentiation. However, there is no evidence that PIs accumulate into preadipocytes and adipocytes and that intra-cellular accumulation is sufficient to alter differentiation. We assessed the effect of six different PIs on the differentiation of cells from four clonal lines. We also studied the capacity of ritonavir to accumulate both into drug-sensitive and drug-resistant cultured adipocytes. METHODS: Adipocyte differentiation of mouse 3T3-F442A, 3T3 L1 and Ob1771 cells as well as embryonic stem cells were investigated at pharmacological concentrations of indinavir, saquinavir, ritonavir, amprenavir, nelfinavir and lopinavir. We used a sensitive ELISA to determine intracellular concentration of ritonavir from 3T3-L1 and Ob1771 preadipocytes. RESULTS: Nelfinavir and lopinavir inhibited adipocyte differentiation whereas amprenavir was ineffective. Indinavir, saquinavir and ritonavir inhibited differentiation of 3T3-L1 and 3T3-F442A cells but did not alter differentiation of either Ob1771 or embryonic stem cells. We showed that ritonavir accumulated in preadipocytes and fully differentiated 3T3-L1 adipocytes as a function of its extracellular concentration. Although Ob1771 cells were resistant and 3T3-L1 cells were sensitive to ritonavir, the drug accumulated to similar levels in both cases. CONCLUSIONS: Protease inhibitors inhibit adipocyte differentiation depending on the cell model used. We showed for the first time that ritonavir accumulates into preadipocytes and adipocytes, suggesting a direct effect on intracellular targets. However, intracellular accumulation was clearly not sufficient as Ob1771 cells remained resistant to the inhibitory effect of ritonavir. PMID- 14523275 TI - Phase I/II trial of intravenous recombinant interleukin-2 in HIV-infected children. AB - OBJECTIVES: To define the tolerated dose of recombinant interleukin-2 (rIL-2) in HIV-infected children (part A), and to determine the safety and immunologic effects of the tolerated rIL-2 dose in a cohort of HIV-infected children (part B). DESIGN: Open-label, dose-escalation. SETTING: Multiple center study. SUBJECTS: Twenty HIV-infected children, aged 3-12 years. INTERVENTION: In part A six subjects received 1 x 10(6) IU/m2 and four subjects received 4 x 10(6) IU/m2 rIL-2 by continuous intravenous infusion for 5 days every 8 weeks for three cycles. In part B 10 different subjects received 1 x 10(6) IU/m2 for 5 days every 8 weeks for six cycles. MAIN OUTCOME MEASURES: Toxicity, CD4 cell count and percentage, and viral load. RESULTS: The tolerated dose of rIL-2 was 1 x 10(6) IU/m2. The most common side effects were fever and vomiting. Of 10 subjects enrolled in part B of the study, five discontinued rIL-2 therapy for a variety of reasons, most related to administration of study drug. Comparable rises in CD4 cell count and percentage were observed in each of the treatment arms. Six cycles of rIL-2 therapy did not appear to be better than three cycles with respect to improvement of CD4 parameters. Transient rises in plasma HIV-1 RNA levels were detected in some subjects. CONCLUSIONS: These results suggest that rIL-2 therapy can raise CD4 cell counts and percentages in some HIV-infected children, although a high proportion of HIV-infected children may have to discontinue intravenous therapy because of drug- or administration-related toxicity. Controlled trials of rIL-2 in this patient population are warranted. PMID- 14523276 TI - Risk of severe hepatotoxicity associated with antiretroviral therapy in the HIV NAT Cohort, Thailand, 1996-2001. AB - OBJECTIVE: To examine rates and predictors of severe hepatotoxicity with combination antiretroviral therapy in a developing country setting: the eight HIV NAT randomized controlled trials in Thailand. METHODS: All patients (n = 692) received at least two nucleoside reverse transcriptase inhibitors; 215 also received a non-nucleoside reverse transcriptase inhibitor (NNRTI) and 135 also received a protease inhibitor. Severe hepatotoxicity was defined as an increase in alanine aminotransferase (ALT) level to five times the upper limit of normal and an increase of at least 100 U/l from baseline. Liver function tests were available at baseline and weeks 4, 8, 12, 24, 36 and 48. Hepatitis B virus (HBV) and hepatitis C virus (HCV) testing was performed on stored serum. RESULTS: Mean age was 32.3 years; 52% were male, 11% had Centers for Disease Control and Prevention category C HIV disease at baseline, and 22% had received prior antiretroviral therapy. Prevalence of HBV, HCV and HBV/HCV coinfection was 8.7%, 7.2%, and 0.4%, respectively. Incidence of severe hepatotoxicity was 6.1/100 person-years [95% confidence interval (CI), 4.3-8.3/100]. In multivariate analysis, predictors of severe hepatotoxicity were HBV or HCV coinfection, and NNRTI-containing therapy. Incidence of severe hepatotoxicity was particularly high among patients receiving nevaripine (18.5/100 person-years; 95% CI, 11.6 27.8) and nevirapine/efavirenz (44.4/100 person-years; 95% CI, 12.1-113.7). CONCLUSIONS: Incidence and risk factors for severe hepatotoxicity appear similar among these Thai patients to those in other racial groups. Development of standardized antiretroviral therapy regimens for developing country settings should consider potential toxicity and capabilities for monitoring of toxicity. PMID- 14523277 TI - Successful implementation of a low-cost method for enumerating CD4+ T lymphocytes in resource-limited settings: the ANRS 12-26 study. AB - OBJECTIVE: To evaluate the feasibility and the relevance of the implementation of an alternative technique to flow cytometry (FC) for enumerating CD4 T cells (Dynabeads; Dynal Biotech, Oslo, Norway), based on quantifying CD4 T cells by epifluorescent microscopy following their isolation using anti-CD4 monoclonal antibody-coated magnetic beads. DESIGN: International multi-center study. Five consecutive runs of dual CD4 T-lymphocyte enumeration by both techniques in six sites in five countries of West Africa. METHODS: A total of 657 pairs of values of CD4 cell counts were generated by 43 technicians by both FC (TruCount; Becton Dickinson Immunocytometry Systems, San Jose, California, USA) and Dynabeads from blood samples obtained from 301 HIV-infected patients, seen in one (n = 112), two (n = 61), three (n = 75), four (n = 40) or five (n = 13) occasions. RESULTS: The correlation coefficient between the results of the two techniques was 0.89. The overall systematic difference between Dynabeads and FC was -16 x 10(6) cells/l (P < 10(-4)). The median difference was insignificant (+7.5 cells) for CD4 cell counts below 200 x 10(6) cells/l and increased with CD4 levels. Patients were consistently classified at the threshold of 200 x 106 cells/l by both methods in 88.7% of cases. Among the 74 discrepant pairs of values, only 31 (4.7%) exhibited a difference of more than 100 x 10(6) cells/l. CONCLUSIONS: Results from Dynabeads and FC were highly correlated. The ability of the alternative method to consistently classify results in agreement with FC, at thresholds of CD4 cell counts relevant for clinical care, was high. The implementation of this low-cost method was easy and successful in the West African context. PMID- 14523278 TI - Case series of acute hepatitis in a non-selected group of HIV-infected patients on nevirapine-containing antiretroviral treatment. AB - OBJECTIVE: To evaluate the characteristics of patients who developed acute clinical hepatitis in an unselected outpatient population. METHODS: Patients who started a nevirapine-containing regimen in the period January 1999-February 2001 and presented with clinical symptoms in accordance with increased transaminase values within 12 weeks of initiation of nevirapine were considered possible cases of clinical hepatotoxicity. Patient characteristics, co-medicated drugs, HIV-1 RNA levels and clinical chemistry parameters were collected from outpatient medical records and clinical medical records. RESULTS: At the defined period, 306 patients started a nevirapine-containing regimen, of whom eight developed an acute hepatitis (2.6%) in a median of 24 days [interquartile range (IQR) 20-25 days]. Transaminases peaked at 28 days (IQR, 27-32 days). Injury pattern was in general mixed-hepatocellular. Withdrawal of the antiretroviral agent led to rapid restoration of transaminase levels and resolution of clinical symptoms. The reason for developing this hepatic reaction was not clear in every case as no specific risk factor(s) covering all patients in this case series could be identified. CONCLUSIONS: It is very important to monitor closely transaminase levels of all patients starting a nevirapine-containing regimen, including patients with no specific characteristics that put them at risk. The rapid onset of the clinical symptoms pleads for transaminase monitoring at a very early stage (i.e., within 2 weeks of initiation) of the nevirapine-containing regimen. PMID- 14523279 TI - Methods and procedures for estimating HIV/AIDS and its impact: the UNAIDS/WHO estimates for the end of 2001. AB - BACKGROUND: The Joint United Nations Programme on HIV and AIDS (UNAIDS) and the World Health Organization (WHO) have produced country-specific estimates of HIV/AIDS biannually since 1997. These estimates are a primary source of information about the extent and spread of the HIV/AIDS epidemic and its impact. The importance of having comparable country-specific estimates of HIV/AIDS is growing as estimates are used to determine how international resources to fight HIV/AIDS will be allocated to countries. OBJECTIVES: This paper describes the procedures and process used to make the 2001 round of UNAIDS/WHO estimates of HIV/AIDS. The paper focuses on the different approaches used to make estimates of prevalence in countries with generalized and low-level and concentrated epidemics as well as on new curve-fitting software that was developed to produce epidemic curves for each country. In addition, it presents the assumptions used (e.g. survival from infection to death, the rate of mother-to-child transmission) that are required to derive estimates of incidence and mortality in adults, as well as prevalence, incidence and mortality in children. CONCLUSION: The paper describes the general process by which the estimation and modelling procedures have been refined and improved over time. The paper also discusses the limitations and weaknesses of the procedures and the data used to make the estimates, and suggests areas where further improvements need to be made. PMID- 14523280 TI - Mortality and progression to AIDS after starting highly active antiretroviral therapy. AB - OBJECTIVES: To examine survival and progression to AIDS among HIV-infected patients after starting highly active antiretroviral therapy (HAART). METHODS: The study population consisted of 3724 patients from the ATHENA observational cohort who initiated HAART. We considered progression to either an AIDS-defining disease or death, distinguishing HIV-related and non-related (including therapy related) deaths. A time-dependent multivariate hazards model was fitted to the patient data and 5-year survival probabilities under various therapy scenarios estimated. RESULTS: A total of 459 patients developed AIDS and 346 died during 12 503 person-years of follow-up. HIV-related mortality decreased from 3.8 to 0.7 per 100 person-years between 1996 and 2000 whereas non-HIV-related mortality did not change (0.4 and 0.9, respectively, P = 0.25). For asymptomatic and symptomatic therapy naive patients younger than 50 years with CD4 counts above 10 x 10(6) and 150 x 10(6) cells/l, respectively, predicted 5-year survival probabilities were above 90% when HAART was used continuously. This limit was 450 x 10(6) cells/l when HAART was used during 20 weeks in each 24 week-period of follow-up, and 110 x 10(6) cells/l when patients delayed initiation of HAART for 1 year after becoming eligible for treatment. CONCLUSIONS: Survival probabilities were high among HIV-infected patients initiating HAART at an early stage of infection. The best therapy strategy is therefore to start HAART at this stage of infection. However, deferring HAART in patients with high CD4 cell counts may be clinically more appropriate given toxicity and adherence problems. The lack of any change in non-HIV-related mortality suggests that toxicity has not yet become a major risk factor for death. PMID- 14523281 TI - Migrants from Sub-Saharan Africa in the Swiss HIV Cohort Study: access to antiretroviral therapy, disease progression and survival. AB - OBJECTIVE: To examine the proportion of migrants from Sub-Saharan Africa entering the Swiss HIV Cohort Study (SHCS) and to compare these participants with participants from Northwestern Europe for access to antiretroviral therapy, progression to AIDS and survival. DESIGN: Prospective national cohort study of HIV-1-infected adults from seven HIV centres in Switzerland. METHODS: Trends in the proportion of participants from Sub-Saharan Africa were followed in 11 872 HIV-infected adults entering the SHCS from 1984 to 2001. Survival methods were used to compare uptake of antiretroviral therapy, survival and progression to AIDS in the 2684 participants from Sub-Saharan Africa and Northwest Europe enrolled from 1997-2001. RESULTS: There was a steady increase in the proportion of Sub-Saharan African participants over time, reaching 11.9% in 1997-2001. These participants were more likely to be younger, female, to have been infected by heterosexual intercourse and had lower CD4 cell counts at presentation. There were no differences between Sub-Saharan Africans and Northwest Europeans in uptake of triple antiretroviral therapy, progression to AIDS or survival up to 48 months after starting treatment. Tuberculosis was the most frequent AIDS-defining event in Sub-Saharan African patients. CONCLUSIONS: There is no evidence that access to potent antiretroviral therapy is influenced by geographic origin of participants. The prognosis of Sub-Saharan African patients on triple therapy is equivalent to that of Northwest European patients. Future research should address wider issues about access to specialist health services for HIV-infected people from Sub-Saharan Africa. PMID- 14523282 TI - Who infects whom? HIV-1 concordance and discordance among migrant and non-migrant couples in South Africa. AB - OBJECTIVES: To measure HIV-1 discordance among migrant and non-migrant men and their rural partners, and to estimate the relative risk of infection from inside versus outside primary relationships. DESIGN: A cross-sectional behavioural and HIV-1 seroprevalence survey among 168 couples in which the male partner either a migrant, or not. METHODS: A detailed questionnaire was administered and blood was collected for laboratory analysis. A mathematical model was developed to estimate the relative risk of infection from inside versus from outside regular relationships. RESULTS: A total of 70% (117 of 168) of couples were negatively concordant for HIV, 9% (16 of 168) were positively concordant and 21% (35 of 168) were discordant. Migrant couples were more likely than non-migrant couples to have one or both partners infected [35 versus 19%; P = 0.026; odds ratio (OR) = 2.28] and to be HIV-1 discordant (27 versus 15%; P = 0.066; OR = 2.06). In 71.4% of discordant couples, the male was the infected partner; this did not differ by migration status. In the mathematical model, migrant men were 26 times more likely to be infected from outside their regular relationships than from inside [relative risk (RR) = 26.3; P = 0.000]; non-migrant men were 10 times more likely to be infected from outside their regular relationships than inside (RR = 10.5; P = 0.00003). CONCLUSIONS: Migration continues to play an important role in the spread of HIV-1 in South Africa. The direction of spread of the epidemic is not only from returning migrant men to their rural partners, but also from women to their migrant partners. Prevention efforts will need to target both migrant men and women who remain at home. PMID- 14523283 TI - HIV infection, antiretroviral therapy, and hepatic function. Emerging epidemiological, pathogenetic, and clinical issues, and their consequences on disease management. PMID- 14523284 TI - Structured interruptions of therapy: looking for the best protocol. PMID- 14523285 TI - Efforts from observational cohorts to transpose results into useful recommendations for clinical management of HIV-infected patients. PMID- 14523286 TI - HIV health experiences among migrant Africans in Europe: how are we doing? PMID- 14523287 TI - Histopathology of the liver in adolescents co-infected with HIV and hepatitis C virus. AB - Chronic hepatitis C is usually a mild disease in children; however, the evolution of children co-infected with HIV is unknown. We report the liver biopsy results of seven adolescents co-infected for over 13 years. According to the Metavir scoring system, liver specimens were classified A1-F1 in three patients and A2-F1 in four. Mild fibrosis progression is mainly explained by young age at hepatitis C virus contamination and the lack of severe immunodeficiency in the study population. PMID- 14523288 TI - The safety, tolerability and effectiveness of generic antiretroviral drug regimens for HIV-infected patients in south India. AB - We investigated the safety, tolerability and effectiveness of locally produced generic highly active antiretroviral therapy (HAART) regimens with a chart review conducted at YRG CARE, a tertiary HIV referral centre in India. A total of 333 patients had been on Indian-manufactured generic HAART for at least 3 months. In this cohort, generic HAART was safe, well tolerated and effective at increasing CD4 T-lymphocyte counts in patients with advanced HIV, comparable to the experience with proprietary HAART. PMID- 14523290 TI - HIV DNA two long terminal repeat circles: observations and interpretations. PMID- 14523289 TI - HIV transmission among gay men through oral sex and other uncommon routes: case series of HIV seroconverters, Sydney. AB - Seventy-five homosexual men with recently acquired HIV were interviewed about their risk behaviour. Fifty-nine reported unprotected anal intercourse, and one shared injecting equipment, with a partner not known to be HIV negative. Of the remaining 15, 11 reported protected anal intercourse. In five of the 15 we judged oral sex to be the most likely source of infection, including three men who had a genital piercing. The possible transmission risk from genital piercing should be investigated. PMID- 14523291 TI - On HIV genetic diversity in Cuba. PMID- 14523293 TI - Expression of P-glycoprotein, multidrug-resistance proteins 1 and 2 in CEM, CEM(VBL), CEM(E1000), MDCKII(MRP1) and MDCKII(MRP2) cell lines. PMID- 14523295 TI - Public health considerations for the use of a first generation HIV vaccine. Report from a WHO-UNAIDS-CDC consultation, Geneva, 20-21 November 2002. AB - SUMMARY: To accelerate the development and future availability of safe, effective and affordable HIV vaccines it is essential to address not only the associated biomedical obstacles, but also the logistic aspects that would guide the introduction and use of those vaccines. It is likely that initial vaccines may only be partially effective, and their public health use will have to be carefully considered. This report summarizes the discussions from a consultation held in Geneva (20-21 November 2002) organized by the World Health Organization (WHO), the joint United Nations Programme on HIV/AIDS (UNAIDS) and the US Centers for Disease Control and Prevention (CDC). The group identified a number of logistic issues that need to be addressed to accelerate the development and future availability of HIV vaccines, and made broad recommendations in four different areas: (a) Vaccine manufacturing and licensing; (b) vaccination acceptability and social marketing; (c) immunisation strategies and delivery; and (d) access and economic issues. The implementation of these recommendations will require the participation of multiple stakeholders in the public and private sector, in industrialized and developing countries. These actions will be essential to ensure widespread and rapid access to HIV vaccines globally, soon after their efficacy is demonstrated in clinical trials. PMID- 14523294 TI - Failures of 1 week on, 1 week off antiretroviral therapies in a randomized trial. AB - BACKGROUND: Scheduled treatment interruptions are being evaluated in an effort to decrease costs and side effects of highly active antiretroviral therapy (HAART). A schedule of 1 week on and 1 week off therapy offers the promise of 50% less drug exposure with continuously undetectable HIV RNA concentration. METHODS: In the Staccato study 600 patients on successful HAART were to be randomized to either continued therapy, CD4-guided therapy, or one week on, one week off therapy. A scheduled preliminary analysis evaluated effectiveness in the 1-week on-1-week-off arm. RESULTS: Of 36 evaluable patients, 19 (53%) had two successive HIV RNA concentrations > 500 copies/ml at the end of the week off therapy, and were classified as virological failure. Most of those who failed took didanosine, stavudine, saquinavir, and ritonavir (11 patients). In these patients, there was no evidence of mutations suggestive of drug resistance, and plasma saquinavir levels were within the expected range. Two of three patients failing on triple nucleotides had drug resistance mutations, but nonetheless responded to reintroduction of triple nucleotide therapy. One of two patients taking nevirapine, and one of eight taking efavirenz, also failed. Both had resistance mutations at the time of failure, but not at baseline. CONCLUSIONS: The 1-week-on 1-week-off schedule, as tested in the Staccato study, showed an unacceptably high failure rate and was therefore terminated. PMID- 14523296 TI - Muscle and strength: an evolution of study. PMID- 14523297 TI - Two years of debilitating pain in a football spearing victim: slipping rib syndrome. AB - INTRODUCTION: Blunt chest trauma can occur in a variety of sports, and lead to rib fractures and less commonly known and diagnosed injuries. We report the case of a 14-yr-old student athlete who was speared (helmet tackled) in a practice scrimmage sustaining a painful injury that eluded diagnosis and treatment for more than 2 yr. METHODS: The case history of pain treatments and radiological evaluations is presented. RESULTS: Ultimately, a definitive diagnosis of "slipping rib syndrome" was achieved through a simple clinical manipulation (the hooking maneuver), combined with a history of symptomatic relief provided with costochondral blockade. Surgical resection of the slipping rib provided total resolution of the problem. CONCLUSION: Very few clinicians are aware either of the syndrome or the maneuver used to diagnose this condition. Although spearing has been outlawed in American football for years, it remains a commonplace occurrence, and sports physicians should be aware of the potential consequences to the victim as well as those to the perpetrator. PMID- 14523299 TI - Traumatic extensor tendon dislocation in a boxer: a case study. AB - An elite collegiate boxer developed extensor tendon subluxation in the small finger of his dominant right hand. He was thought to have a radial sagittal band disruption and was allowed to complete his season with custom padding. Surgical exploration revealed intact sagittal bands with divergent dislocation of the two extensor tendons to the small finger with underlying capsular rupture. These lesions were repaired and he successfully returned to boxing. Soft tissue injuries to the dorsal MCP joint may involve the collateral ligaments, the sagittal bands, the extensor tendons, or the joint capsule. Symptoms usually involve persistent pain, swelling, and tendon subluxation or dislocation. Accurate recognition and treatment is crucial as nonoperative treatment is generally unsuccessful and surgical reconstruction is required for optimal return to function. PMID- 14523298 TI - Venous obstruction in healthy limbs: a model for chronic compartment syndrome? AB - PURPOSE: Chronic exertional compartment syndrome (CECS) in the anterior tibial (AT) compartment is generally believed to be the result of reduced venous blood flow caused by restrictive compartments and increased intramuscular pressures. If this is so, then restricting venous flow in the muscles of healthy subjects during exercise should mimic CECS. METHODS: This hypothesis was tested in 10 control subjects (aged 19-41 yr, five males) with and without external venous occlusion induced by a sphygmomanometer cuff fitted just below the knee and inflated to 80 mm Hg. Twenty CECS patients (20-39 yr, 16 males) were studied without external occlusion. Subjects performed intermittent, isometric maximal voluntary contractions (MVC) of the AT for 20 min (1.6-s contractions, 0.5 duty cycle). MVC, tetanic force (2 s at 50 Hz), muscle thickness (ultrasound imaging), and pain were measured during exercise and 10 min of recovery. RESULTS: Venous occlusion in the controls induced greater pain, fatigue, and increase in muscle thickness (P < 0.01). Initially the patients fatigued more slowly than the occluded controls, but at the end of exercise, the fatigue and pain were similar in these two groups. The controls showed a greater increase in muscle size (P = 0.01). Recovery was similar in all three groups, although the size of the patients' muscles recovered rather more slowly. CONCLUSION: External venous occlusion of the AT muscles in control subjects induces changes very similar to those of CECS patients, although the different time courses indicate that different processes are involved. The AT compartment of CECS patients is capable of distension. PMID- 14523300 TI - Plasma rich in growth factors to treat an articular cartilage avulsion: a case report. AB - INTRODUCTION: The application of an autologous plasma rich in growth factors is beneficial in restoring connective tissues, as shown by clinical evidence in oral surgery and more recently in arthroscopic anterior cruciate ligament reconstruction and two cases of ruptured Achilles tendon in professional athletes. This is attributed to the slow delivery of growth factors from harvested platelets that have been activated by endogenous thrombin promoted by the addition of calcium chloride. PURPOSE: This case report describes a new application of this therapy in the arthroscopic treatment of a large, nontraumatic avulsion of articular cartilage in the knee of an adolescent soccer player. METHODS: After arthroscopic reattachment of the large (>2 cm) loose chondral body in its crater in the medial femoral condyle, autologous plasma rich in growth factors was injected into the area between the crater and the fixed fragment. RESULTS AND CONCLUSION: Despite the extremely poor prognosis of the case, complete articular cartilage healing was considerably accelerated, and the functional outcome was excellent, allowing a rapid resumption of symptom-free athletic activity. This technique opens new perspectives for human tissue regeneration. PMID- 14523301 TI - Repeated treadmill walks affect physiologic responses in children with cerebral palsy. AB - PURPOSE: To determine whether physiologic responses during treadmill walking in children with cerebral palsy (CP) are affected by repeated walking bouts on different days, and whether effects are different at different speeds. METHODS: Three girls and five boys (9.2-15.7 yr, 23.3-64.4 kg) with mild CP received 12-15 min of treadmill walking practice and had their fastest walking speed (FWS) determined during an introductory visit. During each of three subsequent visits (day 1, day 2, day 3), subjects walked for 3 min at 60, 75, and 90% FWS. Resting physiologic measures were taken on day 1. RESULTS: From day 1 to day 3, net ventilation ([OV0312]E), and net heart rate (HR) at 90% FWS decreased by 3.6 L.min-1 and 8 beats.min-1, respectively. There were no differences between day 1 and day 2 or day 1 and day 3 for any other physiologic variable at any speed. Day 3 was less than day 2 for net HR (60% FWS) and, independent of speed, net [OV0312]O2 (per kilogram of body mass and per stride) and net energy expenditure (kJ.min-1). Between-day reliability (R) of physiologic responses was > or = 0.95, except respiratory rate (R = 0.75). Intrasubject, between-day variability for the [OV0312]O2 measures was 7.6-12.9%. CONCLUSION: Because there were no day 1 to day 3 reductions in metabolic variables, day 1 to day 3 reductions at 90% FWS in net HR may reflect decreased emotional stress over time and reductions in net [OV0312]E, an uncoupling of [OV0312]O2 and [OV0312]E. Despite between-day differences, reliable net physiologic and stable net metabolic variables may be collected in subjects with mild CP after one treadmill walking practice session. PMID- 14523302 TI - Exercise and intestinal polyp development in APCMin mice. AB - PURPOSE: Epidemiological evidence suggests that physical activity protects against colon cancer. We previously used a mouse predisposed to intestinal polyps (APCMin) to evaluate this association and found the suggestion of fewer polyps in exercised males but not females. The present study was designed to further explore the potential exercise x sex interaction on polyp development and to begin to look at potential mechanisms. METHODS: Six-week-old APCMin mice (N = 60 males; 60 females) were randomly assigned to one of two groups by sex: treadmill running at 20 m.min-1, 5% grade, 45 min.d-1, 5 d.wk-1 (EX) or nonrunning controls (CON) (N = 30 per group). EX mice ran in running wheels while in quarantine (weeks 0-3), followed by treadmill running weeks 3-8. Body weights were measured weekly. Urine was collected at 5 wk and fasting blood at 7.5 wk. Body composition was measured, serum was frozen, and polyp number and size were measured at sacrifice. RESULTS: EX resulted in lower body weights (P < 0.01) and reduced fat mass (P < 0.01). Fasting glucose was lower in EX (P < 0.01), and leptin was lower in EX (P = 0.05) compared with CON. EX did not affect serum insulin-like growth factor-1 or urinary corticosterone. Total polyp number and size were not statistically different between groups; however, there were fewer jejunal polyps in EX (3.6 +/- 0.7, mean +/- SE) versus CON males (5.2 +/- 0.8; P = 0.04) and an even larger difference when only the consistent runners were kept in the analysis (2.7 +/- 0.5 in EX; P = 0.01). CONCLUSION: Despite favorable changes in body composition, blood glucose, and leptin, 8 wk of running resulted in only minor changes related to polyp development in male but not female APCMin mice. PMID- 14523303 TI - Longitudinal changes in .VO2max: associations with carotid IMT and arterial stiffness. AB - PURPOSE: High levels of cardiorespiratory fitness (.VO2max) are associated with reduced risk of cardiovascular morbidity and mortality. However, little is known to what extent longitudinal changes in .VO2max affect arterial wall thickness and stiffness, i.e., two major risk factors for cardiovascular disease. We therefore investigated the relationship between changes in .VO2max from adolescence (13-16 yr) to adulthood (age 36) and from young adulthood (21-32 yr) to age 36, and carotid intima-media thickness (IMT) and stiffness of the carotid, femoral, and brachial arteries, at age 36. METHODS: Analyses of changes in .VO2max from adolescence to age 36 consisted of 154 subjects (79 women), and from young adulthood to age 36 consisted of a subpopulation of 118 subjects (62 women). Throughout the years, .VO2max was measured directly with a maximal running test on a treadmill. When the subjects had the mean age of 36, carotid IMT and large artery stiffness (distensibility and compliance coefficients) were assessed noninvasively by ultrasound imaging methods. RESULTS: Longitudinal changes in .VO2max were not significantly associated with carotid IMT. Changes in .VO2max were inversely and significantly associated with large artery stiffness. These associations were not uniform throughout the arterial tree, being stronger and independent of changes in other risk factors in the muscular (brachial and femoral) arteries but dependent on and possibly mediated by concomitant changes in HDL cholesterol and body weight in the elastic carotid artery. CONCLUSION: Increases in .VO2max that occur from adolescence up to age 36 are associated with less arterial stiffness. Improving .VO2max by increasing physical activity levels may therefore contribute to a reduction in mortality from cardiovascular disease through decreasing arterial stiffness. PMID- 14523304 TI - Myocardial stress after competitive exercise in professional road cyclists. AB - PURPOSE: Based on the determination of cardiac troponin (cTnT), brain natriuretic peptide (BNP), and echocardiographic measurements, recent investigations have reported myocardial damage and reversible cardiac dysfunction after prolonged endurance exercise in apparently healthy subjects. In the present study, we investigated the myocardial stress reaction in professional endurance athletes after strenuous competitive physical exercise. METHODS: Eleven highly trained male professional road cyclists (age 27 +/- 4 yr; .VO2peak 67 +/- 5 mL.kg-1.min 1; training workload 34,000 +/- 2,500 km.yr-1) were examined. The following parameters were determined before and after one stage of a 5-d professional cycling race: BNP, cTnT (third-generation assay that shows no cross reactivity with skeletal TnT), creatine kinase (CK), creatine kinase MB (CKMB), myoglobin (Myo), and urea. All participants were submitted to a careful cardiac examination including echocardiography and stress ECG. RESULTS: None of the athletes showed pathological findings in the cardiac examination. CK (P < 0.01), CKMB (P < 0.05), and Myo (P < 0.01) were increased after the race. Normal postexercise cTnT levels indicate that the increase in CK, CKMB, and Myo was of noncardiac origin. In contrast, BNP rose significantly from 47.5 +/- 37.5 to 75.3 +/- 55.3 pg.mL-1 (P < 0.01). Pre- and postexercise values of BNP as well as the individual exercise induced increase in BNP were significantly correlated with age (R2 = 0.68, R2 = 0.66, and R2 = 0.58, respectively; P < 0.05). CONCLUSION: Strenuous endurance exercise in professional road cyclists does not result in structural myocardial damage. The rise in BNP in older athletes may reflect a reversible, mainly diastolic left ventricular dysfunction. This needs to be confirmed by larger trials including different intensities, sports, and age groups. PMID- 14523305 TI - Comparison of physical activity in male and female children: does maturation matter? AB - PURPOSE: To investigate whether observed differences in physical activity levels in boys and girls are confounded by biological age differences particularly during the circumpubertal years. METHODS: The physical activity questionnaire for children (PAQ-C) was administered biannually or triannually to 138 (70 boys; 68 girls) Canadian children for seven consecutive years from 1991 to 1997. Participants were 9-18 yr of age. Anthropometric measurements were taken biannually and age at peak height velocity (PHV) determined. Biological age was defined as years from PHV. The data were analyzed using t-tests and random effects models. RESULTS: Level of physical activity decreased with increasing chronological age in both sexes. When aligned on chronological age bands, boys had statistically significantly higher PAQ-C summary scores than girls from 10 through 16 yr of age (P < 0.05). However, when aligned on biological age, sex differences were not apparent, except at 3 yr before PHV. Random effects models of individual growth patterns confirmed these findings. CONCLUSION: Physical activity decreased with increasing chronological age in boys and girls. There were no sex differences in the longitudinal pattern of physical activity when the confounding effects of biological age were controlled except at 3 yr before PHV. PMID- 14523306 TI - Musculoskeletal response to exercise is greatest in women with low initial values. AB - INTRODUCTION: The "initial values" principle of exercise training states those with the lowest initial values of a physiologic system have the greatest capacity for improvement in response to training. We sought to determine whether initial values predicted the musculoskeletal response to training in premenopausal women (N = 31) who participated in a 1-yr program of resistance and jump training designed to improve physical indices of fracture risk. Significant improvements in trochanteric bone mineral density (BMD), hip abductor strength, power, and postural stability occurred in response to training. METHODS: To determine the predictive power of initial values, we performed separate stepwise regression analyses for each variable including the following dependent variables: age, initial value, highest weight lifted during training, and total number of exercise sessions attended. RESULTS: In each case, the initial value was the most significant predictor of percent change in response to training. Initial values explained 15-29% of the variance in the magnitude of the training response. For each unit lower BMD of the greater trochanter (0.01 g.cm-2), the training response was 12% greater. For each unit decrease in initial strength (1 N.m), power (1 W), and stability (1 SI unit), the training response was 1.0%, 0.2%, and 8.0% greater, respectively. When categorized by quartile of initial values, women in the lowest quartile had two- to fivefold greater improvements in musculoskeletal measures than those in the upper quartile. CONCLUSION: Women who began training with the lowest initial values had the greatest improvements in hip BMD, hip abductor strength, leg power, and postural stability. These results support the training principle of initial values and suggest that this training program may be most successful in premenopausal women with lower values of musculoskeletal indices of fracture risk. PMID- 14523307 TI - Deterioration of muscle function after 21-day forearm immobilization. AB - PURPOSE: Although it is well known that immobilization causes muscle atrophy, most immobilization models have examined lower limbs, and little is known about the forearm. The purpose of this study was to determine whether forearm immobilization produces changes in muscle morphology and function. METHODS: Six healthy males (age: 21.5 +/- 1.4, mean +/- SD) participated in this study. The nondominant arm was immobilized with a cast (CAST) for 21 d, and the dominant arm was measured as the control (CONT). The forearm cross-sectional area (CSA) and circumference were measured as muscle morphology. Maximum grip strength, forearm muscle oxidative capacity, and dynamic grip endurance were measured as muscle function. Magnetic resonance (MR) imaging was used to measure CSA, and 31phosphorus MR spectroscopy was used to measure time constant (Tc) for phosphocreatine (PCr) recovery after submaximal exercise (PCr-Tc). Grip endurance was expressed by the number of handgrip contractions at 30% maximum grip strength load. All measurements were taken before and after the immobilization. RESULTS: After the 21-d forearm immobilization, no changes were seen for each measurement in CONT. CSA and the circumference showed no significant changes in CAST. However, maximum grip strength decreased by 18% (P < 0.05), PCr-Tc was prolonged by 45% (P < 0.05), and the grip endurance at the absolute load was reduced by 19% (P < 0.05) for CAST. CONCLUSION: In this model, 21-d forearm immobilization caused no significant changes in forearm muscle morphology, but the muscle function showed remarkable deterioration ranging from 18 to 45%. PMID- 14523308 TI - Targeting the metabolic syndrome with exercise: evidence from the HERITAGE Family Study. AB - PURPOSE: To determine the efficacy of exercise training in treating the metabolic syndrome. METHODS: The sample included 621 black and white participants from the HERITAGE Family Study, identified as sedentary and apparently healthy (no chronic disease or injury). The metabolic syndrome was defined as having three or more risk factors according to the guidelines of the National Cholesterol Education Program, including elevated waist circumference, blood pressure, triglycerides, blood glucose, and low HDL cholesterol. The presence of the metabolic syndrome and component risk factors were determined before and after 20 wk of supervised aerobic exercise training. RESULTS: The prevalence of the metabolic syndrome was 16.9% in this sample (105/621) of apparently healthy participants. Of the 105 participants with the metabolic syndrome at baseline, 30.5% (32 participants) were no longer classified as having the metabolic syndrome after the exercise training. Among the 32 participants who improved their metabolic profile, 43% decreased triglycerides, 16% improved HDL cholesterol, 38% decreased blood pressure, 9% improved fasting plasma glucose, and 28% decreased their waist circumference. There were no sex or race differences in the efficacy of exercise in treating the metabolic syndrome: 32.7% of men, 28.0% of women, 29.7% of black, and 30.9% of white participants with the metabolic syndrome were no longer classified as having the syndrome after training. CONCLUSION: Aerobic exercise training in patients with the metabolic syndrome can be useful as a treatment strategy and provides support for a role for physical activity in the prevention of chronic disease. PMID- 14523309 TI - Orthotic comfort is related to kinematics, kinetics, and EMG in recreational runners. AB - PURPOSE: The purpose of this study was to determine the relationship between differences in comfort and changes in lower extremity kinematic and kinetic variables and muscle activity in response to foot orthoses. METHODS: Twenty-one recreational runners volunteered for this study. Three orthotic conditions (posting, custom-molding, and posting and custom-molding) were compared with a control (flat) insert. Lower extremity kinematic, kinetic, and EMG data were collected for 108 trials per subject and condition in nine sessions per subject for overground running at 4 m.s-1. Comfort for all orthotic conditions was assessed in each session using a visual analog scale. The statistical tests used included repeated measures ANOVA, linear regression analysis, and discriminant analysis (alpha = 0.05). RESULTS: Comfort ratings were significantly different between orthotic conditions and the control condition ([lower, upper] confidence limits; posting: [-3.1, -0.8]; molding: [0.4, 3.4]; and posting and molding: [ 1.1, 1.9]); 34.9% of differences in comfort were explained by changes in 15 kinematic, kinetic, and EMG variables. The 15 kinematic, kinetic, and EMG variables that partially explained differences in comfort classified 75.0% of cases correctly to the corresponding orthotic condition. DISCUSSION: In general, comfort is an important and relevant feature of foot orthoses. Evaluations of foot orthoses using comfort do not only reflect subjective perceptions but also differences in functional biomechanical variables. Future research should focus on defining the relationship between comfort and biomechanical variables for material modifications of footwear, different modes of locomotion, and the general population. PMID- 14523311 TI - Demographic characteristics of elite Ethiopian endurance runners. AB - INTRODUCTION: The dominance of East-African athletes in distance running remains largely unexplained; proposed reasons include favorable genetic endowment and optimal environmental conditions. PURPOSE: To compare the demographics of elite Ethiopian athletes with the general Ethiopian population and assess the validity of reports linking running long distances to school with endurance success. METHODS: Questionnaires, administered to 114 members (male and female) of the Ethiopian national athletics team and 111 Ethiopian control subjects (C) obtained information on place of birth, language, distance and method of travel to school. Athletes were separated into three groups according to athletic discipline: marathon (M; N = 34); 5,000-10,000 m (5-10 km; N = 42); and other track and field athletes (TF; N = 38). Frequency differences between groups were assessed using contingency chi-square tests. RESULTS: Regional distributions of marathon athletes differed from controls (P < 0.001) and track and field athletes (P = 0.013), but not the 5- to 10-km athletes (P = 0.21). The 5- to 10-km athletes also differed from controls (P < 0.001). Marathon athletes exhibited excess from the regions of Arsi and Shewa (M: 73%; 5-10 km: 43%; TF: 29%; C: 15%). The language distribution of marathon athletes differed from all groups (P < 0.001), with a predominance of languages of Cushitic origin (M: 75%, 5-10 km: 52%, TF: 46%, C: 30%). A higher proportion of marathon athletes ran to school (M: 68%; 5 10 km: 31%; TF: 16%; C: 24%) and traveled greater distances. CONCLUSION: Elite endurance athletes are of a distinct environmental background in terms of geographical distribution, ethnicity, and also having generally traveled farther to school, often by running. These findings may reflect both environmental and genetic influences on athletic success in Ethiopian endurance athletes. PMID- 14523310 TI - Knee flexor torque and perceived exertion: a gender and reliability analysis. AB - PURPOSE: The objectives of the present study were to examine gender differences and between-day variability of isometric hamstring muscle peak torque and perceived exertion. METHODS: Subjects included 20 healthy, college-aged male (N = 10) and female (N = 10) volunteers. Each subject completed five maximal voluntary isometric hamstring muscle contractions (MVC), in a prone position, with their knee at 30 degrees flexion. Subjects then completed, in random order, isometric contractions at 10, 20, 30, 40, 50, 60, 70, 80, and 90% of their three highest averaged MVC. Perceived exertion was measured with a modified Borg CR-10 scale after each contraction. Ten randomly chosen subjects were asked to return approximately 1 wk after the initial evaluation to repeat the same procedure. Peak hamstring muscle torque was examined in absolute (N.m), relative (N.m.kg-1), and allometric-scaled (N.m.kg-n) units. Perceived exertion across the contraction intensities was modeled to a power function in order to determine the exponent and proportionality constant. RESULTS: Males generated significantly greater hamstring muscle torque than females in absolute, relative, and allometric-scaled units (P < 0.05). No significant differences in perceived exertion occurred across the submaximal contraction intensities between females and males, nor for the derived exponents and proportionality constants. Perceived exertion ratings were observed to be significantly lower (P < 0.05) across the contraction intensity range on the second day. CONCLUSIONS: The major findings demonstrated that perceived exertion did not differ between healthy young female and male adults, despite males generating significantly more hamstring muscle torque, and perceived exertion ratings decreased at similar relative contraction intensity levels across testing days. PMID- 14523312 TI - Does lower-body BMD develop at the expense of upper-body BMD in female runners? AB - PURPOSE: Evidence suggests that exercise plays an important role in stimulating site-specific bone mineral density (BMD). However, what is less well understood is how these benefits dissipate throughout the body. Hence, the purpose of the present study was to compare the levels of, and the correlation between, BMD recorded at 10 sites in female endurance runners, and to investigate possible determinants responsible for any inter-site differences observed. METHODS: Repeated measures ANOVA was used to compare the BMD between sites and factor analysis was used to describe the pattern of intersite correlations. Allometric ANCOVA was used to identify the primary determinants of bone mass and how these varied between sites. RESULTS: The ANOVA and factor analysis identified systematic differences in BMD between sites, with the greatest BMD being observed in the lower-body sites, in particular the legs. An investigation into the possible mechanisms responsible for these differences revealed "distances run" (km.wk-1) as a positive, and "years of training" as a negative determinant of bone mass (P < 0.001). However, the effect of a number of determinants varied between sites (P < 0.05). Specifically, the ANCOVA identified that running further distances resulted in higher bone mass in the arms and legs. In contrast, training for additional years appeared to result in lower bone mass in the arms and lumbar spine. Calcium intake was also found to be positively associated with bone mass in the legs but negatively associated at all other sites. CONCLUSIONS: A combination of running exercise and calcium intake would appear to stimulate the bone mass of women endurance runners at lower-body sites but at the expense of bone mass at upper-body sites. PMID- 14523313 TI - Spine bone mineral density increases in experienced but not novice collegiate female rowers. AB - PURPOSE: There is evidence that rowing exercise targets the spine, but the dose of exercise required to build bone is poorly understood. To further explore this topic, we studied the bone response at the spine in novice and experienced female collegiate rowers over their 6-month competitive season. We hypothesized that, with rowing strokes similar between groups during training, experienced rowers would produce greater force at the spine than novices and thus, gain more bone mineral density (BMD). METHODS: Subjects included 16 experienced rowers (21.2 +/- 1.2 yr) who had been rowing 26 +/- 10 months and 19 novice rowers (19.5 +/- 0.8 yr) who had been rowing three months and 14 controls (19.2 +/- 1.6 yr). BMD was assessed by DXA at baseline and after the 6-month competitive season. During the season, all rowers participated in the same training program and took approximately the same number of strokes per training session (1000-1200 repetitions). On the 2000- and 6000-m rowing ergometer tests experienced rowers had faster times, indicating that their power output, and thus force production, was greater than novices. RESULTS: In ANCOVA, adjusting for body mass index, age, and initial BMD, the experienced rowers demonstrated a 2.5% increase at the spine that was significantly different than that of the novice rowers, but BMD change scores in the rowing groups were not different than controls. CONCLUSION: Because the number of strokes (repetitions) was similar between rowing groups during training, the higher power output in experienced rowers produced higher forces at the spine over the 6-month period that resulted in gains in spine BMD. These results support the theory that force magnitude is a key variable in osteogenesis. PMID- 14523314 TI - Valgus knee motion during landing in high school female and male basketball players. AB - PURPOSE: The purpose of this study was to utilize three-dimensional kinematic (motion) analysis to determine whether gender differences existed in knee valgus kinematics in high school basketball athletes when performing a landing maneuver. The hypothesis of this study was that female athletes would demonstrate greater valgus knee motion (ligament dominance) and greater side-to-side (leg dominance) differences in valgus knee angle at landing. These differences in valgus knee motion may be indicative of decreased dynamic knee joint control in female athletes. METHODS: Eighty-one high school basketball players, 47 female and 34 male, volunteered to participate in this study. Valgus knee motion and varus valgus angles during a drop vertical jump (DVJ) were calculated for each subject. The DVJ maneuver consisted of dropping off of a box, landing and immediately performing a maximum vertical jump. The first landing phase was used for the analysis. RESULTS: Female athletes landed with greater total valgus knee motion and a greater maximum valgus knee angle than male athletes. Female athletes had significant differences between their dominant and nondominant side in maximum valgus knee angle. CONCLUSION: The absence of dynamic knee joint stability may be responsible for increased rates of knee injury in females but is not normally measured in athletes before participation. No method for accurate and practical screening and identification of athletes at increased risk of ACL injury is currently available to target those individuals that would benefit from neuromuscular training before sports participation. Prevention of female ACL injury from five times to equal the rate of males would allow tens of thousands of young females to avoid the potentially devastating effects of ACL injury on their athletic careers. PMID- 14523315 TI - Regulating oxygen uptake during high-intensity exercise using heart rate and rating of perceived exertion. AB - PURPOSE: The purpose of this study was to comparatively evaluate the use of heart rate (HR) or rating of perceived exertion (RPE) in eliminating the slow component of oxygen uptake (.VO2) during high-intensity aerobic exercise. METHODS: Nine sedentary males (age = 23.9 +/- 4.6 yr, height = 177.4 +/- 10.1 cm, weight = 75.28 +/- 12.95 kg) completed three 15-min submaximal exercise cycle ergometer tests based on: 1) constant power output (PO) corresponding to 75% .VO2max (PO75), 2) HR corresponding with 75% .VO2max (HR75), and 3) RPE response corresponding with 75% .VO2max (RPE75). .VO2, HR, RPE, and blood lactate concentration [La-] were measured during all tests. Data were analyzed using repeated measures analysis of variance, and post hoc means comparisons were performed using a Fisher's LSD test. RESULTS: End-exercise .VO2 was significantly higher than the respective 3-min .VO2 for the PO75 and RPE75 tests, but not the HR75 test. End-exercise .VO2 was significantly greater for the PO75 test than both the RPE75 and HR75 tests, but there was no significant difference between end-exercise .VO2 for the RPE75 and HR75 tests. End-exercise HR and RPE were significantly higher for the PO75 test than both the RPE75 and HR75 tests. There were no significant differences between the RPE75 and HR75 tests for end-exercise HR or end-exercise RPE. CONCLUSION: Results suggest using both HR and RPE are effective at reducing the slow component of .VO2 that occurs during high intensity exercise. PMID- 14523316 TI - Coagulation and fibrinolytic responses to manual versus automated snow removal. AB - PURPOSE: The purpose of this study was to assess coagulation and fibrinolytic responses to snow removal. METHODS: Thirteen healthy male subjects (age = 26 +/- 5 yr, height = 179.0 +/- 7.0 cm, weight = 78.7 +/- 16.1 kg, .VO2max = 54.7 +/- 8.9 mL.kg-1.min-1) underwent maximal treadmill stress testing (TM), 10 min of snow shoveling (SS), and 10 min of snow removal using an automated snow thrower (ST). Blood was collected immediately before and after each test and analyzed for von Willebrand Factor antigen (vWF:ag), tissue plasminogen activator (tPA) antigen, and plasminogen activator inhibitor-1 (PAI-1) activity. Data were analyzed using a two-factor repeated-measures analysis of variance. RESULTS: vWF:ag significantly increased during TM (84.7 +/- 21.7% normal preexercise, 149.0 +/- 45.6% normal postexercise) but not SS or ST. TM resulted in significant increases in tPA antigen (6.54 +/- 2.76 ng.mL-1 preexercise, 21.39 +/- 10.56 ng.mL-1 postexercise) and both SS and TM caused significant reductions in PAI-1 activity (SS = 15.1 +/- 3.8 AU.mL-1 preexercise, 13.2 +/- 4.3 AU.mL-1 postexercise; TM = 15.3 +/- 3.6 AU.mL-1 preexercise, 10.5 +/- 5.3 AU.mL-1 postexercise). Postexercise PAI-1 activity was significantly lower for TM versus SS. tPA antigen was unchanged after SS and ST, and PAI-1 activity was unchanged after ST. CONCLUSION: vWF:ag is unchanged after self-paced snow shoveling and automated snow removal in young, healthy males. Snow shoveling acutely increases fibrinolytic potential in this population, although not to the degree observed after maximal treadmill exercise. PMID- 14523317 TI - Water turnover and changes in body composition during arduous wildfire suppression. AB - INTRODUCTION: Our lab has recently documented the total energy expenditure during arduous wildfire suppression using the doubly labeled water methodology. The elevated rates of isotopic elimination indicate an arduous working environment that may often compromise energy balance and overall hydration. PURPOSE: The purpose of this investigation was to determine the effects of arduous wildfire suppression on water turnover and changes in body composition in wildland firefighters (WLFF). METHODS: WLFF (N = 14) were studied during a 5-d period of arduous fire suppression work. A comparison group (N = 13) of recreationally active college students (RACS) was also studied. Water turnover was measured from rates of 2H elimination (rH2O). Urine osmolality, specific gravity, and skinfold measures were also collected. RESULTS: WLFF demonstrated a decrease in nude body weight (pre = 71.9 +/- 10.4 kg, post = 70.9 +/- 10.2 kg, P = 0.0001) and total body water (pre = 42.9 +/- 7.2 kg, post = 42.0 +/- 6.7 kg, P = 0.0046). RACS maintained total body water and body weight during the experimental period. Isotope (2H2O) dilution demonstrated that rH2O was significantly higher for the WLFF (rH2O = 6.7 +/- 1.4 and 3.8 +/- 1.0 L.24 h-1 for the WLFF and RACS, respectively). CONCLUSION: These results demonstrate an arduous work environment that threatens hydration, energy balance, and perhaps normal glycogen status. PMID- 14523318 TI - Triaxial accelerometry to evaluate walking efficiency in older subjects. AB - PURPOSE: We tested the suitability of triaxial accelerometry to evaluate walking efficiency in older subjects. METHODS: First, we verified the accuracy to estimate the oxygen consumption rate (.VO2, mL.min-1) from the total impulse (Itotal, N.min-1), the square root of summed accelerations of each direction, during graded walking on a flat ground in 13 male and 27 female older subjects (61 +/- 6 yr, mean +/- SD). Second, to examine the effects of endurance/resistance training on walking efficiency, we assessed the relations of maximal isometric knee extension force (Fmax, N.m), maximal walking velocity (Vmax, m.min-1), and three-dimensional impulses (Ix, anterior-posterior; Iy, mediolateral; Iz, vertical) in 13 male and 40 female older subjects (62 +/- 7 yr) before and after 6 and 9 months of training. RESULTS: The following analyses were performed in all the data from the male and female groups. First, .VO2 was highly correlated with Itotal (r = 0.958, P < 0.0001) over the range of 250-2200 mL.min 1. Second, Fmax and Vmax increased by 48 +/- 7% (P < 0.001) and 21 +/- 2% (P < 0.001), respectively, after 9 months of training. Ix/Itotal and Iy/Itotal increased by 18 +/- 2% (P < 0.001) and 10 +/- 2%, respectively, after 9 months of training (P < 0.001), whereas Iz/Itotal decreased by 14 +/- 2% (P < 0.001). Vmax was negatively correlated with Iz/Itotal (r = -0.522, P < 0.0001) while positively correlated with Ix/Itotal (r = 0.561, P < 0.0001) and Iy/Itotal in the pooled data from before, after 6 and 9 months of training. Similarly, the product of Vmax and body weight was positively correlated with Fmax (r = 0.633, P < 0.0001). CONCLUSIONS: These results suggest that increased Fmax improved walking efficiency by increasing energy utilization in the anterior posterior/mediolateral directions while decreasing energy loss in the vertical direction. PMID- 14523319 TI - Technical variability of the RT3 accelerometer. AB - PURPOSE: To evaluate the technical performance of the RT3 triaxial accelerometer. METHODS: Twenty-three RT3 accelerometers were subjected to a specific vibration along each sensitive axis in isolation, using a motorized vibration table that produced frequencies of 2.1, 5.1, and 10.2 Hz, respectively. Data were analyzed for frequency and axis effects and inter- and intra-instrument variability. RESULTS: ANOVA showed a frequency by axis interaction (F2.1,36.8 = 19.9, P < 0.001). Post hoc tests revealed the Y axis count to be significantly higher than the X and Z axes counts at 5.1 and 10.2 Hz. There was no difference in counts between axes at 2.1 Hz. Interinstrument coefficients of variation (CV) decreased as frequency increased (21.9 to 26.7% at 2.1 Hz, 6.3 to 9.0% at 5.1 Hz, and 4.2 to 7.2% at 10.2 Hz). The intraclass correlation (ICC) between RT3s was 0.99, regardless of the axis. Intra-instrument CV also decreased as frequency increased (2.1 to 56.2%, 0.3 to 2.5%, and 0.2 to 2.9% at 2.1, 5.1, and 10.2 Hz, respectively. CONCLUSION: There were no differences in counts recorded on the X, Y, and Z axes at 2.1 Hz; however, the counts recorded along the Y axis were significantly higher than the counts at the X and Z axes at 5.1 and 10.2 Hz. Due to large coefficients of variation for both inter- and intra-instrument variability at 2.1 Hz, testing the inter- and intra-instrument variability of the accelerometers before use is recommended. PMID- 14523320 TI - Accuracy and reliability of 10 pedometers for measuring steps over a 400-m walk. AB - PURPOSE: The purpose of this study was to determine the accuracy and reliability of the following electronic pedometers for measuring steps: Freestyle Pacer Pro (FR), Kenz Lifecorder (KZ), New Lifestyles NL-2000 (NL), Omron HJ-105 (OM), Oregon Scientific PE316CA (OR), Sportline 330 (SL330) and 345 (SL345), Walk4Life LS 2525 (WL), Yamax Skeletone EM-180 (SK), and the Yamax Digi-Walker SW-701 (DW). METHODS: Ten males (34.7 +/- 12.6 yr) (mean +/- SD) and 10 females (43.1 +/- 19.9 yr) ranging in BMI from 19.8 to 33.6 kg.m-2 walked 400-m around an outdoor track while wearing two pedometers of the same model (one on the right and left sides of the body) for each of 10 models. Four pedometers of each model were assessed in this fashion. The actual steps taken were tallied by a researcher. RESULTS: The KZ, NL, and DW were the most accurate in counting steps, displaying values that were within +/-3% of the actual steps taken, 95% of the time. The SL330 and OM were the least accurate, displaying values that were within +/-37% of the actual steps, 95% of the time. The reliability within a single model (Cronbach's alpha) was >0.80 for all pedometers with the exception of the SL330. The intramodel reliability was exceptionally high (>0.99) in the KZ, OM, NL, and the DW. CONCLUSION: Due to the variation that exists among models in regard to the internal mechanism and sensitivity, not all pedometers count steps accurately. Thus, it is important for researchers who use pedometers to assess physical activity to be aware of their accuracy and reliability. PMID- 14523321 TI - Female athlete issues for the team physician: a consensus statement. PMID- 14523322 TI - Chronic exertional compartment syndrome: muscle changes with isometric exercise. PMID- 14523324 TI - Double-contrast MR colonography: in vivo experimental study in an animal model. AB - BACKGROUND: We evaluated the performance of double-contrast MR colonography in detecting colorectal tumors experimentally induced by chemical substances in the large bowel of the rat. MATERIAL/METHODS: Eight Sprague-Dawley rats developed 67 chemically induced tumors of the large bowel. The animals were imaged with a double-contrast MR technique at 0.5T, using a head coil for both signal transmission and reception. Prior to MR acquisition, room air was insufflated into the rats' large bowel. Unenhanced and contrast-enhanced T1, proton-density and T2 sequences were obtained in the axial and coronal planes. Autopsy was performed immediately after MR examination. MR images were interpreted by consensus of two observers, and the results were compared with post-mortem data. RESULTS: Sensitivity (60% vs. 93%), specificity (63% vs. 81%), positive (85% vs. 93%) and negative (30% vs. 81%) predictability, and global diagnostic accuracy (60% vs. 89%) were obtained for all tumors (n=67) and for the group of tumors larger than 1.5 cm (n=43), respectively. No tumors less than 1.5 cm were detected. CONCLUSIONS: Double-contrast MR colonography is a promising modality in detecting colorectal neoplasms larger than 1.5 cm in an animal tumor model. PMID- 14523323 TI - GW274150 inhibits nitric oxide production by primary cultures of rat proximal tubular cells. AB - BACKGROUND: Production of nitric oxide (NO) subsequent to expression of inducible NO synthase (iNOS) contributes to the development of ischemic renal injury and inflammation. Here we investigate the effects of GW274150, a potent, long-acting and highly selective inhibitor of iNOS activity, on NO production by primary cultures of rat proximal tubular cells (PTC). MATERIAL/METHODS: Pure populations of PTC were isolated from the cortex of kidneys obtained from male Wistar rats using a combination of collagenase digestion, sieving and Percoll centrifugation. Confluent PTC cultures were incubated for 1-24 h with MEM, interferon-gamma (IFN gamma, 100 microg/ml), bacterial lipopolysaccharide (LPS, 10 microg/ml) in combination after which NO production was determined. PTC were also incubated with IFN-gamma (100 microg/ml) and LPS (10 microg/ml) and increasing concentrations of GW274150 or L-N6-(1-iminoethyl)lysine (L-NIL) (10 nM-1 mM) for 24 h after which nitrite levels in the incubation medium were measured. RESULTS: IFN-g and LPS in combination produced a significant, time-dependent increase in NO production. Both GW274150 and L-NIL produced a significant and concentration dependent inhibition of NO production. However, GW274150 was markedly more potent (EC50 approximately 100 nM) than L-NIL (EC50 approximately 10 microM). CONCLUSIONS: GW274150 inhibits NO production by primary cultures of PTCs and may therefore be useful in conditions associated with nitrosative stress of the kidney. PMID- 14523325 TI - Reduced presence of tissue-repairing cells in wounds combined with whole-body irradiation injury is associated with both suppression of proliferation and increased apoptosis. AB - BACKGROUND: The goal of our study was to examine the effects of whole body irradiation injury on tissue-repairing cells, so as to elucidate the possible mechanisms of reduced repopulation of tissue-repairing cells. MATERIAL/METHODS: The quantity of tissue-repairing cells in wounds from simple incision injury (WBI) (group S) and combined with WBI injury (group C) was measured histologically. The proliferation and apoptosis of tissue-repairing cells in wounds were determined using immunohistochemical staining for proliferation cell nuclear antigen (PCAN) and by TUNEL assay. Fibroblasts were isolated from the wounds, and their proliferation and apoptosis were detected in vitro. Some key molecules involving in proliferation (cyclin E and cyclin dependent kinase 4) and apoptosis (bcl-2 and bax) were also determined, so as to interpret the mechanisms of the alteration of tissue repairing cells. RESULTS: The quantity of tissue repairing cells in group C was significantly less than in group S. The PCNA contents in wounds of group C were significantly less than in group S, whereas the apoptosis of tissue-repairing cells in wounds of group C increased significantly as compared to group S. The expression of the molecules involved in the mediating of proliferation and apoptosis coincided with the proliferation and apoptosis of tissue repairing cells in wounds. CONCLUSIONS: The quantity of tissue repairing cells in wounds combined with WBI injury is significantly less than in simple incision injury. This is associated with both suppression of proliferation and increased apoptosis. PMID- 14523326 TI - Effect of ionic strength of buffer on the measurement of erythrocyte electrophoretic mobility. AB - BACKGROUND: Measurement of the electrophoretic mobility (EM) of biological cells provides information on the electrical properties of the cell surface. Different research groups have used different electrophoretic techniques. It is well known that the osmolality of the buffer used during an experiment exerts a major impact on EM, particularly in biological cells. In previous experiments either high or low ionic strength electrophoresis buffers were used with an arbitrary preference. The aim of this study is to investigate which buffer is more useful for EM measurements, and hence to give greater insight into the results of such experiments. MATERIAL/METHODS: Blood was drawn from healthy volunteers. Two different buffers were prepared: low ionic and high ionic. Twenty molarity steps were prepared by mixing the buffers in different ratios. Measurements were made using a Zetasizer. RESULTS: Our data confirmed that the EM fell as ionic strength increased. However, the relationship was biphasic: the first ten points were linear with a steep negative slope and the second set also linear but with a much smaller slope. CONCLUSIONS: EM studies usually have one of two extreme aims: either to compare the inherent surface electrical properties of cells from different sources at the same ionic strength, or to study the influence of different ionic environments on the surface electrical properties of cells from the source. Our data suggest that in the first instance it is better to work at high ionic strength. In the second, low ionic strength is preferable. PMID- 14523327 TI - Norepinephrine infusion increases interleukin-6 in plasma and cerebrospinal fluid of brain-injured rats. AB - BACKGROUND: Significantly increased plasma and CSF IL-6 levels reflect underlying tissue damage following clinical and experimental traumatic brain injury (TBI). Catecholamines, used under clinical conditions to maintain adequate cerebral perfusion pressure, induce a sustained IL-6 release. Thus an additional elevation in IL-6 could aggravate brain edema in the acute posttraumatic phase. We studied the changes in plasma and cerebrospinal fluid (CSF) IL-6 levels 4 and 24 hours after experimental TBI and assessed possible time-dependent effects of norepinephrine infusion on IL-6 and brain edema. MATERIAL/METHODS: Paired plasma and CSF IL-6 measured at 4 and 24 hours following TBI (n=10) were compared to levels in non-traumatized rats (n=5). In a placebo-controlled trial, 20 brain injured male Sprague-Dawley rats were randomized to receive norepinephrine or NaCl for 90 minutes at 4 or 24 hours after TBI. Plasma IL-6 was measured before, during, and after the infusion period. One hour after stopping the infusion, CSF IL-6 and hemispheric swelling were determined. RESULTS: During the first posttraumatic day, plasma and CSF IL-6 levels were significantly increased compared to non-traumatized rats, reaching the highest values at 24 hours (p<0.05). Norepinephrine infusion significantly increased plasma IL-6 at 7 and 27 hours after TBI; IL-6 was significantly elevated in CSF only at 7 hours (p<0.05). Brain edema was not aggravated. CONCLUSIONS: The norepinephrine-induced increase in plasma and CSF IL-6 suggests that concomitant norepinephrine administration needs to be considered when interpreting systemic and local changes in IL-6 levels in TBI patients. PMID- 14523328 TI - Embolic cerebral ischemia in carotid surgery: a model for human embolic stroke? AB - BACKGROUND: If carotid endarterectomy (CEA) is a clinical model of embolic stroke, then it should be possible to determine the relevance of embolus size and of the total quantity of embolic material (embolus load) for ischemic stroke. MATERIAL/METHODS: Thirty-one patients underwent a total of 33 CEAs. Diffusion weighted imaging (DWI) was performed on the day before and after CEA. A new hyperintense signal on the postoperative DWI was considered a new ischemic lesion. Intraoperatively detected microembolic signals (MES) were quantified (in cm) by multiplying embolus velocity by the embolus presence within the Doppler sample volume. With this relative index, emboli were classified into small (<0.56 cm), medium-sized (0.56-1.12 cm) and large (1.12-1.70 cm). RESULTS: For each category of embolus size, embolus load (in cm/minute) was expressed as the sum of all quantified emboli divided by minutes of CEA duration. By logistic regression analysis, the occurrence of a new ischemic lesion was related to an embolus load with medium-sized emboli (without lesion, 0.029 cm/minute; with lesion, 0.157 cm/minute; p=0.018), and with large emboli (without lesion, 0.015; with lesion, 0.055; p=0.038), but not to an embolus load with small emboli or the largest embolus per CEA. CONCLUSIONS: The leading reason for cerebral ischemia in CEA is multiple embolism with emboli of medium and large size. This conceptual model may also apply to carotid artery disease. PMID- 14523329 TI - Haemostatic gene polymorphisms in young indian asian subjects with acute myocardial infarction. AB - BACKGROUND: The relationship between haemostatic gene polymorphisms and arterial disease remains unclear. Much of the evidence gathered so far has been obtained from small heterogeneous studies, resulting in inconsistencies. This study focuses on the South African Indian population which not only represents the largest Indian population outside the Indian subcontinent, but also constitutes a genetically discrete group in whom a high incidence of coronary heart disease occurs. MATERIAL/METHODS: We investigated the relationship between polymorphisms in the Factor V (Leiden), prothrombin (20210 GgA) and thrombomodulin (Ala455Val) genes in patients with a myocardial infarction (MI) <45 years of age (n=195) and in unaffected siblings (n=107) and unrelated healthy race-matched individuals drawn from the same community (n=300). RESULTS: The Factor V Leiden mutation was found to occur with a frequency of less than 1%, while the prothrombin 20210 GgA polymorphism was not observed in any of the individuals in this study. In contrast, the variant thrombomodulin Val allele occurred with a frequency similar to that reported in Caucasians. The frequency of this allele in both patients with MI and their siblings was marginally higher than in healthy controls, but the difference did not reach statistical significance. However, in patients who smoked, the thrombomodulin variant allele occurred significantly more frequently than in the non-smoking group (p=0.044). CONCLUSIONS: The Leiden Factor V and prothrombin 20210 GgA polymorphisms have no value in disease association studies in the Indian Asian population. In smokers, the thrombomodulin Ala455Val variant allele emerges as a significant risk factor for coronary heart disease. PMID- 14523330 TI - Indolyl-3-acryloylglycine (IAG) is a putative diagnostic urinary marker for autism spectrum disorders. AB - BACKGROUND: Autism is a heterogeneous pervasive developmental disorder with a poorly defined aetiology and pathophysiology. There are indications that the incidence of the disease is rising but still no definitive diagnostic biochemical markers have been isolated. Here we have addressed the hypothesis that urinary levels of trans -indolyl-3-acryloylglycine (IAG) are abnormal in patients diagnosed with autism spectrum disorders (ASD) compared to age-matched controls. MATERIAL/METHODS: Urine samples were collected on an opportunistic basis and analysed for IAG concentration (normalised against creatinine content to account for changes in urinary volume) using reversed phase HPLC with UV detection. RESULTS: Statistical analysis (Mann-Whitney tests) showed highly significant increases (p=0.0002) in the levels of urinary IAG in the ASD group (median 942 microV per mmol/L of creatinine [interquartile range 521-1729], n=22) compared to asymptomatic controls (331 [163-456], n=18). Detailed retrospective analysis showed that gender (boys 625 microV per mmol/L of creatinine [294-1133], n=29; girls 460 [282-1193], n=11: P=0.79) and age (control donor median 10 years [8 14], n=15; ASD median 9 years [7-11] n=22: P=0.54) were not significantly correlated with IAG levels in this non-blinded volunteer study. CONCLUSIONS: Our results strongly suggest that urinary titres of IAG may constitute an objective diagnostic indicator for ASD. Mechanisms for the involvement of IAG in ASD are discussed together with future strategies to address its specificity. PMID- 14523332 TI - Office treatment of congenital ankyloglossia. AB - BACKGROUND: Congenital oral adhesions may pose both aesthetic and functional disturbing ailments to wary parents. Most of these as we have experienced, are benign, easily cured and may be treated as soon as possible in the office. A local survey elucidated that these youngsters are usually advised to wait until one year of age and then taken into the operating room and incision of the adhesion is performed under general anesthesia. We assessed the benign nature of this ailment and relative avascularity of the tissue involved and concluded that with minimal risk an office procedure under local anesthesia can replace current practice. We also found that most referrals with this condition present with the lowest grade of severity of ankyloglossia, amenable to a very brief intervention. MATERIAL/METHODS: During the period 1998-2002 we diagnosed nineteen congenital lesions in thirteen patients. All children were treated in a community clinic setting using electrocautery under local anesthesia. Surgical success was defined as significant improvement in the ability to protrude the tongue outwards beyond the gums and teeth. Gingival adhesions were judged by release of soft tissue adhesions. RESULTS: Tongue Surgical success was accomplished in all cases with minimal discomfort and without complications. In one single case the previous functional limitation was not relieved. CONCLUSIONS: Our experience indicates that office-based electrocautery dissection is an efficacious economical and safe treatment of mild congenital oral adhesions. We recommend this method as therapy of choice for such lesions. PMID- 14523331 TI - Serum procalcitonin concentration as a negative predictor of serious bacterial infection in acute sickle cell pain crisis. AB - BACKGROUND: A pilot study was designed to determine if serum procalcitonin levels would assist in the diagnosis of severe bacterial infections in patients presenting to an emergency department (ED) with acute sickle cell pain crisis and evidence of acute inflammatory response. MATERIAL/METHODS: Prospective single cohort study evaluating measured procalcitonin levels in patients with sickle cell pain crisis and evidence of acute inflammation. Acute inflammation was defined as fever (>38 degrees C) and/or elevation in the white blood cell count (>4000 above baseline) and tachycardia (heart rate >100). Procalcitonin was measured using a semi-quantitative monoclonal antibody test. Patients were followed clinically to determine if procalcitonin has predictive value in excluding severe bacterial infections. RESULTS: Twenty four subjects were enrolled and completed the study. Sixteen had levels 0.5 ng/ml or less, two had levels 0.5 to 2 ng/ml, one had a level of 2 but less than 10 ng/ml, and four had levels 10 ng/ml or greater. All subjects with documented infections at presentation had procalcitonin levels > or =2.0 ng/ml. The sensitivity of the test in this study sample was 1, and the specificity was 0.95 (95% CI, 0.75 0.99). CONCLUSIONS: A serum procalcitonin less than 2 ng/ml appears to have good negative predictive value in excluding serious bacterial infections in patients that present with acute sickle cell pain crisis and evidence of acute inflammatory response. Further study is needed to investigate if procalcitonin has positive predictive value in identifying patients with serious bacterial infections in this patient population. PMID- 14523333 TI - Parent-child concordance on reported barriers to helmet use by children. AB - BACKGROUND: We determined the agreement between parents and their school-age children on reported barriers to bicycle helmet use. MATERIAL/METHODS: We conducted a cross-sectional survey in a 2-county Central Texas region with no helmet use legislation. Parent-child pairs, selected through a randomized, multi stage sampling procedure and pre-matched by unique identifiers, were surveyed independently with self-administered questionnaires. Parent-child concordance was assessed using Kappa statistic. RESULTS: Of 1,987 surveys distributed, children returned 1,203 (61%) while parents returned 636 (53% of child respondents). Parents included 31% college graduates and 33% reported annual income of >55,000 dollars. Children included 60% whites, 16% blacks, and 16% Hispanics. The most popular reasons cited by children as barriers to helmet use were not having one (34%), being a safe rider (31%), and helmets messing up hair (18%). Good concordance was found on reported child bicycle riding (parent=88.0% vs. child=88.0%; K=0.64) and child helmet use the last time child rode a bicycle (parent=25.1% vs. child=28.0%; K=0.54). While good concordance was also found for reported helmet ownership (parent=31.9% vs. child=29.9%; K= 0.50), poor concordance was found for other barriers to helmet use including helmets looking ugly (parent=6.5% vs. child=13.4%; K=0.38) and messing up hair (parent=5.6% vs. child=15.2%; K=0.36). CONCLUSIONS: Although parents may be generally aware of their children's bicycle riding and helmet use habits, they may not be fully aware of the reasons why their children may not use a helmet. Public education and interventions on helmet promotion need to focus on parents. PMID- 14523334 TI - Host and risk factors for gestational trophoblastic disease: a hospital-based analysis from India. AB - BACKGROUND: Gestational trophoblastic disease (GTD) adversely affects pregnancy by the development of Hydatidiform mole and placental site gestational tumor. GTD is a discrete pool of epidemiological and clinicopathological entities. Therefore, every geographical region should be studied separately. The Indian subcontinent is under-reported for this disease. This paper analyses the data from a large hospital. MATERIAL/METHODS: 92 patients with GTD were analyzed in the period from 1995 to 1999. RESULTS: The incidence of GTD was 1.31 per 1000 live births and one per 967 pregnancies irrespective of outcome. 93.5% of the patients had moderate to severe vaginal bleeding as presenting symptom, while routine antenatal ultrasonography revealed GTD in 6.5% completely asymptomatic patients. Among the major risk factors were age over 30 (p=6.3 x 10(-3), RR=2.7), previous abortion (p<0.001, RR=3.9) and multigravida (p=4.9 x 10(-6), RR=4.1). Early detection and treatment on established lines can achieve 95.7% complete response and 6.5% partial response. 17 out of 21 invasive moles (80.9%) were found with multiple abortions and mechanical interference in form of check curettage, which probably implicates repeated mechanical interference in the conversion of a mole into an invasive one. CONCLUSIONS: The present study explores the status of established host-related risk and prognostic factors in the Indian context. In cases of invasive mole, the role of repeated mechanical interference in the form of check curettage needs further evaluation. PMID- 14523335 TI - Metabotrophic potential of neurotrophins:implication in obesity and related diseases? AB - In addition to their stimulatory action on neuronal differentiation and survival, the neurotrophins nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) improve glucose and lipid metabolism and control energy balance and feeding behavior. These latter activities are referred to here as the metabotrophic potential of neurotrophins. We recently reported that circulating NGF and BDNF levels are reduced in the metabolic syndrome and in acute coronary syndromes, and that the tissue content of NGF is reduced in atherosclerotic coronary arteries. Thus we hypothesize that a metabotrophic deficit due to reduction of neurotrophin availability may be implicated in the pathogenesis of obesity and related metabolic diseases, such as metabolic syndrome, type 2 diabetes, and atherosclerosis. The metabotrophic deficit hypothesis also considers metabolism-related beneficial effects exerted by other neurotrophic factors, particularly ciliary neurotrophic factor, leukemia inhibitory factor, and bone morphogenetic proteins. PMID- 14523336 TI - Assessment of left ventricular mass and geometry by cardiac single photon emission computed tomography. AB - BACKGROUND: Left ventricular (LV) hypertrophy is an independent predictor of increased morbidity and mortality. However, whether assessment of LV mass and geometry by cardiac single photon emission computed tomography (SPECT) is feasible remains unclear. MATERIAL/METHODS: We compared LV mass and geometry measured by cardiac SPECT to standard echocardiography in thirty consecutive patients (female 73%n mean age 22.64+/-17 years) who had both cardiac SPECT and echocardiography done less than 1 week apart. RESULTS: There was good correlation between the LV mass (r=0.73, p <0.001) and relative wall thickness (r=0.50, p<0.01) derived by cardiac SPECT to those of standard echocardiography. Using Bland-Altman method, the 95% confident interval of agreement between cardiac SPECT and standard echocardiography ranged from -52 g (5th percentile) to 124 g (95th percentile, mean difference =36+/-45) for LV mass and from -1 (5th percentile) to -0.6 (95th percentile, mean difference =-0.8+/-0.1) for relative wall thickness. CONCLUSIONS: Assessment of left ventricular mass and geometry using cardiac SPECT is feasible and may provide prognostic information beyond measurement of perfusion and function. PMID- 14523337 TI - Imaging of mild traumatic brain injury using 57Co and 99mTc HMPAO SPECT as compared to other diagnostic procedures. AB - BACKGROUND: Traumatic brain injury (TBI) is usually assessed with the Glasgow Coma Scale (GCS), CT and EEG. TBI can result from either the primary mechanical impact or secondary (ischemic) brain damage, in which calcium (Ca) plays a pivotal role. This study was undertaken to compare the applicability of SPECT using 57Co as a Ca-tracer in patients with mild traumatic brain injury. MATERIAL/METHODS: 8 patients with mild TBI (GCS 15) were clinically examined and studied with EEG, neuropsychological testing (NPT) and SPECT within 2 days post TBI. After i.v.-administration of 37 MBq (1 mCi) 57Co (effective radiation dose 0.34 mSv x MBq(-1); 1.24 rem x mCi(-1); physical half-life 270 days, biological half-life 37.6 h), single-headed SPECT (12 h pi) was performed, consecutively followed by standard 925 MBq (25 mCi) Tc-99m HMPAO SPECT. RESULTS: In 6 of the 8 patients, baseline NPT and SPECT showed focal abnormalities in the affected frontal and temporal brain regions, which were in good topographical accordance. CT and EEG did not detect (structural) lesions in any of these cases. CONCLUSIONS: Single-headed 57Co-SPECT is able to show the site and extent of brain damage in patients with mild TBI, even in the absence of structural lesions. It may confirm and localize NPT findings. The predictive value of 57Co SPECT should be assessed in larger patient series. PMID- 14523338 TI - The use of glyceryl trinitrate (GTN) in the treatment of chronic anal fissure in children. AB - BACKGROUND: 0.2 per cent topical glyceryl trinitrate (GTN) ointment heals up to two-thirds of chronic anal fissures in adults although many patients experience troublesome headaches. This double blind randomised pilot study assessed the efficacy and side effects of 0.1 and 0.05 per cent GTN ointment in treatment of chronic anal fissures in children. MATERIAL/METHODS: 15 consecutive children with chronic anal fissures were randomised to receive either 0.05 or 0.1 per cent GTN ointment applied topically twice daily for eight weeks. Clinical review was undertaken at weeks 4 and 8 and questions relating to symptoms and the incidence of headache were asked. RESULTS: The median age of the 15 (8 male) patients was 6 years (range 3-13). There were 5 anterior and 10 posterior fissures. The median duration of symptoms was 9 (3-30) months. 13/15 (86.7%) patients were taking laxatives at the time of referral. Fissure healing was complete at 8 weeks in all seven patients who received 0.05% and in 5/8 (62.5%) patients using the 0.1% ointment (Fisher's exact test, not significant). One patient from each arm experienced headaches during the first week of the trial which resolved without treatment and did not affect compliance. There were no other side effects. CONCLUSIONS: Topical glyceryl trinitrate ointment is effective in healing chronic anal fissures in children. Healing rates and side effect profile are comparable when either 0.05 or 0.1 per cent ointment is used. PMID- 14523339 TI - The effect of metronidazole in treating human fascioliasis. AB - BACKGROUND: The aim of this study was to determine the effect of metronidazole in patients who did not cure after treatment with triclabendazole, in Guilan (Northern Province of Iran). MATERIAL/METHODS: Patients, who passed fasciola egg in stool and had positive serum anti fasciola antibody (ELISA), at least three months after treatment with triclabendazole, were enrolled and received 1.5 g/day metronidazole orally for three weeks. Two months and 12 months after end of therapy, stool examination in 3 consecutive days and serum anti fasciola antibody were performed. Frequency of patients with negative serology for fasciola and/or absence of fasciola egg in stool were determined. Chi-square test was used and P value <0.05 was considered significant. RESULTS: Forty-six patients, 26 females and 20 males, were enrolled with mean (+/-SD) age of 34.6(+/-9.8) years. Three patients excluded because of drug side effect and poor compliance. Two months after end of therapy, stool exam became negative in 35 patients and in 31 patients became negative both in serology and stool examination. (Difference in response to treat between age groups and genders was not significant). All patients with abdominal pain became pain free after therapy. Most frequent side effects were metallic taste in 14 (30.4%), headache in 8 (17.4%) and nausea in 6 (13%). 12 months after end of therapy, 28 out of 35 patients were examined again and all were negative both in serology and egg in stool examination. CONCLUSIONS: Metronidazole, 1.5 g/day for 3 weeks, seems to be an effective, available, well tolerated alternative for treatment of human fascioliasis. PMID- 14523340 TI - Influence of estrogens on neurodegenerative processes. AB - Gonadal steroids are known to be of prime importance in the normal maintenance of brain function. Data from epidemiological studies suggest that the decline in estrogen following menopause may increase the risk of neurodegenerative diseases. There is accumulating epidemiological evidence for neuroprotection by estrogens against neurodegenerative diseases, such as Alzheimer's disease. Although it is unclear whether estrogen may be effective in preventing further cognitive decline in women who already have Alzheimer's disease, several studies indicate that estrogen replacement improves the cognitive performance of postmenopausal women. Despite the somewhat controversial data from human studies, experimental investigations in different animals have shown that estrogen is neuroprotective. Although estrogens are known to exert several direct effects on neurons, the cellular mechanisms involved in the neuroprotective effects of estrogen are still unclear. Estrogen causes most of its neuroprotective effects by means of direct binding to specific nuclear receptors. The anti-oxidant effects of estrogens and activation of different membrane-associated intracellular signaling pathways or neurotrophic cross talk through the signal cascade shared with neurotrophic factors could also contribute to neuroprotection. In this article, we outline the evidence supporting a direct role of estrogen in neuronal survival, and PMID- 14523341 TI - Can genetic material retrotransposition be a cause of pathology accompanying cerebral aneurysm rupture? AB - Our considerations were prompted by observation of patients who underwent surgery due to cerebral aneurysm rupture. Elevated anti-HIV antibody titers were detected in such patients. The above condition was not observed either in subjects with intracranial hemorrhages from cerebral angiomas. The observed titers remained certainly below the cut off level indicating contact with HIV. We analyzed the subpopulations of peripheral blood lymphocytes both in the surgery patients and in the control group represented by blood collected from blood donors. On the other hand, the proliferation potential of circulating lymphocytes in the blood of patients with subarachnoid hemorrhages due to cranial aneurysm ruptures was found to be decreased. Dislocation of transposons, so-called 'unauthorized recombination' is known to occur within the framework of genetic code pathology. Endogenous human viruses have been described, and they may be involved in the development of autoimmune disorders. Additionally, viruses involved in autoimmune processes, which may be 'identical' with retrotransposons, have been described. Moreover, there are studies demonstrating that AIDS is caused by retrotransposition of genetic code material fragments. Our considerations are substantiated by ultrastructural analyses of material coming from the gyrus rectus cortex fragments, resected in patients who underwent clipping of a ruptured aneurysm of the anterior communicating artery. We demonstrated in neuronal chromatin the presence of a molecule ca. 80 nm in diameter, corresponding with its size to retroviruses or genetic material molecules with altered substructure. The authors suggest a new mechanism of development of neurological deficits in patients with ruptured cerebral aneurysms. PMID- 14523342 TI - A meta-analysis of 20 case-control studies of the glutathione S-transferase M1 (GSTM1) status and colorectal cancer risk. AB - BACKGROUND: Glutathione S-transferase M1 is an important phase II enzyme involved in the detoxification of many environmental carcinogens. It has been postulated that individuals with GSTM1 deficiency have increased susceptibility to carcinogens and are more likely to develop cancer. GSTM1 status has been extensively studied as a colon cancer risk factor, although published studies have produced conflicting results. To re-examine this controversy, we have undertaken a meta-analysis investigating the relationship of GSTM1 status and colon cancer risk. MATERIAL/METHODS: Odds ratio was employed to evaluate the risk of colon cancer and GSTM1 status. To take into account the possibility of heterogeneity across the studies, a statistical test for heterogeneity across the studies was performed. The summary odds ratios were assessed by calculating a weighted average of odds ratios for all of the studies. RESULTS: The pooled odds ratio of colon cancer risk associated with the GSTM1 deficiency is 0.99 (95% CI: 0.91-1.07). If pooling studies were based on the genotyping method, the overall odds ratio of colon cancer risk associated with the GSTM1 deficiency showed no difference. Moreover, the relationship of GSTM1 deficiency and colon cancer risk was also conducted on the specific ethnic groups and tumour site. CONCLUSIONS: The results of our meta-analysis does not support the hypothesis that GSTM1 alone is an important risk factor for colon cancer, and suggests that GSTM1 status has no effect on the risk of developing colon cancer. PMID- 14523345 TI - 17-beta estradiol down regulates ganglionic microglial cells via nitric oxide release: presence of an estrogen receptor beta transcript. AB - OBJECTIVES: In earlier studies we have demonstrated that 17-beta-estradiol and an estrogen cell surface receptor can be found on various human cells where they are coupled to nitric oxide release. We also demonstrated the presence of estrogen signaling in Mytilus edulis ganglia. In the present report, we sought to determine a function for these ganglionic estrogen receptors, transcending a reproductive role for estrogen. MATERIAL & METHODS: Ganglionic microglial egress from excised pedal ganglia was examined microscopically following pharmacological treatments designed to determine a role for 17-beta-estradiol in microglial regulation via nitric oxide. Additionally, we examined the tissue by RT-PCR and sequence analysis for the estrogen receptor beta gene. RESULTS: In ganglia incubated with varying concentrations of 17-beta-estradiol-BSA there is a significant drop in microglial egress at the 24 hour observation period (58.7 +/- 7.4 vs. 17-beta-estradiol-BSA exposed = 14.7 +/- 1.5; P<0.01), which can be antagonized by tamoxifen and significantly diminished by L-NAME, a nitric oxide synthase inhibitor. By RT-PCR and sequence analysis Mytilus edulis pedal ganglia was found to express a 266 bp fragment of the estrogen receptor-beta gene, which exhibits 100% sequence identity with the human counterpart. CONCLUSION: These data suggest that 17-beta-estradiol-BSA is working on estrogen cell surface receptors since 17-beta-estradiol-BSA does not enter the cytoplasm and that these receptors are coupled to constitutive nitric oxide release. This study demonstrates that 17-beta-estradiol can down regulate microglial fMLP induced activation and activation following ganglionic excision. PMID- 14523346 TI - The presence of 17-beta estradiol in Mytilus edulis gonadal tissues: evidence for estradiol isoforms. AB - OBJECTIVES: In earlier studies, we demonstrate that 17-beta -estradiol and an estrogen cell surface receptor can be found on various human cells, i.e., vascular endothelial, monocytes, and granulocytes, where they are coupled to nitric oxide release. We further demonstrated this phenomenon in the marine mussel Mytilus edulis ganglionic tissues. In the present report we sought to determine if estrogen can be found in M. edulis reproductive tissues. MATERIAL & METHODS: We determined the presence of 17-beta -estradiol via high pressure liquid chromatography (HPLC) and radioimmunoassay (RIA) in the animals gonads. This substance was further identified via nanoelectro-spray ionization quadrupole time of flight mass spectrometry (Q-TOF-MS). RESULTS: 17-beta -estradiol was identified and quantified in Mytilus gonads. Interestingly, we also determined that estradiol isoforms also were present in this tissue. CONCLUSION: These data demonstrate that 17-beta-estradiol and an estradiol isoform is present in M. edulis gonadal tissues, suggesting that they have functions related to reproduction. This further suggests that estrogen's association with reproductive activities has a long evolutionary history and that this association began in invertebrates. PMID- 14523347 TI - The role of the estrogen in neuroprotection: implications for neurodegenerative diseases. AB - In trying to rectify the differences in the risk, onset, and progression of neurodegenerative diseases between men and women, the gonadal hormone estrogen has been the primary focus of investigation for many years. Although this gender difference may encompass disparate and overlapping reasons, estrogen and signaling events mediated by its receptor have been shown to be neuroprotective in a number of neurodegenerative disease models such as Alzheimer's, Parkinson's, and Schizophrenia. Although data from human studies remains highly controversial, a large body of research findings suggests that this hormone plays a pivotal role in retarding and preventing the formation of neurodegenerative diseases through its receptor. By activating common intracellular signaling pathways and initiating "cross talk" with neurotrophins, estrogen plays an influential role in neuronal survival from injuries induced by ischemia or other environmental insults. Gaining a better understanding of these estrogen receptor mediated neuroprotective mechanisms may lead to new therapeutic strategies for the treatment or prevention of neurodegenerative diseases. PMID- 14523348 TI - Is ADHD adaptive or non-adaptive behavior? AB - The evolutionary approach to the issue of ADHD derives from the assumption that what is regarded as a pathological phenomenon today was once an adaptive response to the conditions of life in the ancestral stages of human development. The paper argues against this conception on the basis of the clinical picture of ADHD. The author believes that in the previous "natural" conditions the ADHD syndrome was even more of a maladaptation than in the "protective" conditions of present-day life. PMID- 14523349 TI - Order and disorder in the brain function. AB - The interest in studying the brain electrical activity as a function of the development of intelligence has been spurred by the need to understand how the brain responds to environmental information. The description of sleep in mentally retarded children reveals deviant patterns of the EEG-spindles and of the eye movement activity (REM sleep) when compared to normal children. The patterns may be considered as a valuable index of mental function. According to experimental evidence, the distribution of the eye movements of sleep appears either as random or ordered. The latter are altered in the mentally handicapped in whom the appearance out of chaos, of the order which is needed for intelligence and memory to function, is altered. The sleep signs are redundant as from birth. Their pattern is also related to the psychomotor development of the infant. If their distribution remains random, or appears in long uninterrupted sequences of waves as in epilepsy, intelligence does not develop. A similar strategy appears to function in the foetus when nature organizes the structures that will lead to the development of intelligence. The eye movement patterns of sleep change in the pregnant women as a function of term and resemble those of premature babies of a similar gestational age. They also change as a function of the menstrual cycle and more generally as a function of age. The hypothesis that attention is the diurnal equivalent of REM sleep is discussed. Attempts at modelling the eye movement patterns of REM sleep as a function of near zero gravity environments have been made. 1) By means of a Montecarlo simulation using the semi Markov model during the Spacelab 1 flight. 2) With the method of the single and multiple g-phase transition analysis of the strange attractor dimension (d) during parabolic flights. The implication of the latter for the neural processes involved in learning is that the central nervous system can preserve intact, from input to output, over a period of several days, all the information it receives 3) The relation between spindles and eye movements has also been viewed by a quantum approach which is another medium between the information and the way of describing it. PMID- 14523350 TI - Etiopathogenesis of Graves' disease. AB - Graves' disease is an autoimmune disorder, caused by thyroid-stimulating antibodies, which bind to and activate the thyrotropin receptor on thyroid cells, inducing the synthesis and release of thyroid hormones. It is a polygenic and multifactorial disease that develops as a result of complex interaction between genetic susceptibility and environmental and/or endogenous factors. Graves' disease differs from other autoimmune diseases of the thyroid by specific clinical features, including hyperthyroidism, vascular goitre, ophthalmopathy and -less commonly--infiltrative dermopathy. This article discusses current theories, regarding the etiology and pathogenesis of Graves' disease, including possible predisposing factors, autoimmune aspects of Graves' disease, ophthalmopathy, and dermopathy. PMID- 14523351 TI - The steroid-responsive hiccup reflex arc: competitive binding to the corticosteroid-receptor? AB - Hiccups occurring secondary to high-doses of corticosteroids are a well recognized problem in the field of neurosurgery. Numerous reports of oral, intravenous and intraarticular corticosteroids inducing hiccups exist in the literature. To date, there is only one case of anabolic steroids inducing hiccups. We now present a case of a patient who underwent a suboccipital craniotomy for resection of a cerebellar pontine angle meningioma. Postoperatively the patient was on high doses of Decadron and Oxandrin, an anabolic-anticatabolic agent used to combat the deleterious effects of corticosteroids. The patient suffered intractable hiccups postoperative day one, resistant to Thorazine. Oxandrin was discontinued to assess the possibility of a anabolic steroid-induced singultus. The hiccups resolved within 24 hours. This report validates the previous report on anabolic steroids inducing hiccups and exemplifies the ability for steroids as a class, due to there backbone structural homology, to induce function even as competitive inhibitors. PMID- 14523352 TI - The effect of prophylactic melatonin administration on reperfusion damage in experimental testis ischemia-reperfusion. AB - AIM: Torsion of testis, which is a urologic emergency case, is generally treated by surgical detorsion procedure. However, the resulting reperfusion and both ipsilateral and contralateral testis damage caused thereby are important problems. This study aims at investigating the administration of prophylactic melatonin in order to reduce free radical damage that is caused due to reperfusion after experimental testis torsion-detorsion procedure. MATERIAL AND METHOD: The rats used in the study were allocated to four groups, each containing 10 rats. Rats in Group I had six hours of torsion (ischemia) followed by orchiectomy. Rats in Group II had six hours of torsion, then received melatonin (10 mg/kg/IM) and after that had detorsion (reperfusion). Rats in Group III had detorsion after six hours of torsion and serum physiologic administration (the same volume as melatonin). Torsion and/or detorsion procedures were not applied in Group IV (control). Ipsilateral, contralateral testis and plasma Malondialdehyde (MDA) levels were determined in all groups. RESULTS: Tissue and plasma MDA levels in the group which had detorsion were found to be significantly higher than those in the group that had orchiectomy only (p<0.02). Ipsilateral and contralateral testis MDA levels were identified to be significantly lower in the group receiving prophylactic melatonin in comparison to the group receiving serum physiologic (p<0.02). Plasma and contralateral MDA levels correlated positively with MDA levels identified in ipsilateral testis in all groups (rs=+0.89, n=40, p=0.000). CONCLUSION: In cases where testis torsion is identified, administration of melatonin just before torsion may reduce local and systemic free radical damage. PMID- 14523354 TI - Reduction of night/day difference in melatonin blood levels as a possible disease related index in schizophrenia. AB - OBJECTIVE: Aim of this study was to verify if a simple index as night-day plasma MLT level variation is able to confirm the existing data on circadian melatonin alterations in schizophrenia and if a relationship to disease itself instead of the actual clinical state can be suggested. SETTING AND DESIGN: Ten consecutively admitted male schizophrenic inpatients were examined. METHODS: The blood samples for melatonin were collected at 3.00 a.m. and 15.00 p.m. and consequently calculated the values of Delta () (MLT h.3.00- MLT h.15.00). We divided the sample into two subgroups: < 30 pg/ml and > 30 pg/ml taking 30 pg/ml as an arbitrary value, based on literature data, that should indicate a physiologically correct value of. RESULTS: The 70% of the sample was under the 30 pg/ml value of (13.61 +/- 4.0) or was lacking of the characteristic circadian pattern of MLT secretion, whilst the 30% of the sample was over the 30 pg/ml value of (83.60 +/- 16.34) or was in presence of the characteristic circadian pattern of MLT secretion (p=.0001). No correlation was found between values and the scale and subscales scores for the assessment of psychopathology. MAIN FINDINGS: The data confirm the lack of the characteristic circadian pattern of MLT secretion in schizophrenics. CONCLUSION: The absence of significant correlation between night / day melatonin level differences and actual psychopathology variables should indicate that the suppression of is mostly related to the disease and independent from the clinical state. A neuroleptic-treatment effect cannot be excluded so far. PMID- 14523353 TI - The effects of music and visual stress on testosterone and cortisol in men and women. AB - OBJECTIVES: The aims of the present study were to examine sex-related differences in testosterone (T) and cortisol (C) changes with music listening and visual stress. SETTING AND DESIGN: Saliva T and C concentrations were measured in 88 healthy college students (44 males and 44 females). These subjects were placed in one of 4 different conditions: (1) 30 min of listening to music, (2) 30 min of listening to music with visual stress (documentary film without sound including violent scenes), (3) 30 min of visual stress without music, and (4) 30 min of silence. METHODS: All subjects provided two saliva samples, one collected before intervention and the other after intervention. T and C levels were assessed by radio immuno assay (RIA). RESULTS: There was a significant difference between the sexes in the way music affected T. Music decreased T in males, whereas it increased T in females. As for C, no sex-related differences were found under any of the conditions studied. C decreased with music and increased under other conditions. THE MAIN FINDINGS: Our data suggests that the effects of music and stress on T differ between males and females. CONCLUSION: Further investigation is necessary to evaluate the relationships between music and other substances, the effect of degree of preference and hormonal changes not only during music listening but also during music plays and creation. PMID- 14523355 TI - The effect of estradiol, but not progesterone, on the production of cytokines in stimulated whole blood, is concentration-dependent. AB - OBJECTIVES: The purpose of this study was to determine the effects of estradiol and progesterone on interferon-gamma (IFN-gamma), interleukin (IL)-12, IL-10 and tumor necrosis factor-alpha (TNF-alpha) productions in polyclonal activators (phytohemagglutinin+lipopolysaccharide)-stimulated whole blood cultures. METHODS: Nineteen healthy males and females volunteered in the study. Blood samples were drawn, diluted, and cultured for 24h with different concentrations of estradiol, progesterone or hydrocortisone and then PHA+LPS was added for another 24 h. The supernatant, then, was harvested and assayed for IL-12 p70, IFN-gamma, IL-10 and TNF-alpha. RESULTS: At preovulatory concentrations, estradiol enhanced significantly IFN-gamma, IL-12 and IL-10, but not TNF-alpha, production levels and reversed the suppressive effect of hydrocortisone in PHA+LPS stimulated whole blood. While IL-10 levels kept increasing at pregnancy estradiol concentrations, IFN-gamma, IL-12 levels and IFN-gamma/IL-10 ratio decreased significantly. No effect of progesterone on IL-12 p70, IFN-gamma, IL-10 and TNF production levels was observed. CONCLUSIONS: The present study shows that those pregnancy estradiol concentrations (and higher) enhance the production of IL-10 and reduce IL-12, IFN gamma levels and IFN-gamma/IL-10 ratio in stimulated whole blood cells. Because of the known IL-10 inhibitory actions on T helper (Th) 1 cells and monocytes/macrophages, these high IL-10 levels keep Th2 cytokines favored during pregnancy and may be useful in shifting Th1-mediated autoimmune diseases towards non-pathogenic Th2 pathway. PMID- 14523356 TI - Effects of central and peripheral administration of leptin on pain threshold in rats and mice. AB - OBJECTIVE: This study was planned to investigate the effects of exogenous leptin on the pain threshold. METHODS: Adult male Wistar rats weighing 250-300 g and mice weighing 25-30 g were used in this study. Leptin was intracerebroventricularly (i. c. v.) injected in a dose of 3.5 micro g/rat. Mice were intraperitoneally (i. p.) injected with leptin in a dose of 25 micro g/mouse. Control animals were injected with the respective vehicle. The pain threshold test was performed using hot plate analgesia meter. The experiments were performed during the day and at night. The data were statistically analysed by Mann-Whitney U test. Level of significance was set at p<0.05. RESULTS: During the day, there were no significant changes in hot plate latencies half an hour after i.c.v. injection of vehicle or leptin in the control and leptin-treated rats, respectively. At night, like during the day, i.c.v. injection of neither vehicle nor leptin caused any significant change in pain sensitivity. In mice, i.p. injection of leptin decreased latencies significantly (p<0.05) during the day and at night. Thus, leptin caused an increase in pain sensitivity during the day and at night. CONCLUSION: These results clearly demonstrated that leptin has a decreasing-effect on pain threshold if it is peripherally administered in mice. PMID- 14523357 TI - Effects of mental stress on cardiovascular and endocrine response in Air Force Academy cadets. AB - OBJECTIVES: Pilots are exposed to the "stress of flight" and the chronically activated stress response may play an important role in circulatory system disease progression. We studied the effect of an experimental mental stress on cardiovascular and neuroendocrine activity, in Air Force Academy cadets, before and after one month intensive course. DESIGN: Nine cadets were submitted to a psychometric evaluation before Stroop test (ST), including Minnesota Multiphasic Personality Inventory, State and Trait Anxiety Inventory (STAI X1 and X2 form) and Reaction Scheme Test. After ST, subjects completed a STAI X1 form. Heart rate (HR), systolic and diastolic blood pressure (SBP and DBP) were monitored and serum prolactin (PRL), growth hormone (GH), adrenocorticotropin (ACTH) and cortisol levels were determined, during ST. The protocol was repeated at the end of the course. RESULTS: No significant differences were evident in PRL, GH and ACTH levels. Cortisol concentrations were significantly higher before the course. ST did not modify hormone secretion. ST induced a significant and reproducible elevation of HR and SBP. Basal HR, SBP and DBP values were significantly elevated after the course. Two subjects presented a particular psychometric profile, a different cardiovascular response to ST and did not pass the course. CONCLUSIONS: acute mental stress significantly activated cardiovascular response without modifying endocrine responses, in relation to the psychological profiles. A significant increase of cardiovascular parameters was present after the course, with enhanced dichotomy between the endocrine and sympathetic system, suggesting a careful evaluation and follow-up for circulatory system diseases in cadets. PMID- 14523358 TI - Testosterone and gonadotropin levels in men with dementia. AB - OBJECTIVES: Sex steroids such as testosterone and estradiol might protect the brain against Alzheimer's disease (AD). We previously found lower levels of testosterone in men with AD compared with controls. We wanted to assess levels of pituitary gonadotropins that regulate sex steroid levels, to determine whether primary or secondary hypogonadism was responsible for low levels of testosterone in cases. METHOD: We included 45 men with AD (McKhann, 1987), 15 men with other types of dementia and 133 elderly controls from the Oxford Project to Investigate Memory and Ageing. Gonadotropins (follicle stimulating hormone or FSH and luteinizing hormone or LH), sex hormone binding globulin (SHBG, which determines the amount of free testosterone) and testosterone were measured using enzyme immunoassays. RESULTS: We found no difference in average LH (8.7 +/- 9 UI/L), FSH (13 +/- 17 UI/L) or SHBG (44 +/- 18 nmol/L) levels between AD cases and controls. Similar to our earlier findings, testosterone levels were significantly lower in men with AD (13 +/- 6 nmol/L) compared with controls (17 +/- 8, O.R. = 0.92, 95% C.I. = 0.87 to 0.97, p<0.005). Results were unchanged when controlled for age, SHBG and gonadotropin levels. CONCLUSION: Although normal, the levels of gonadotropins were inappropriately low for the levels of testosterone. Our results support a preliminary conclusion that secondary hypogonadism occurs in men with AD. This could be a consequence of brain degeneration. This is contrary to an earlier study (Bowen, 1999) that found raised levels of gonadotropins in cases with AD, suggesting primary hypogonadism. Our cohort was younger than theirs and gonadotropin levels increase with age. We are enlarging our data set to investigate whether primary hypogonadism occurs in older cases with AD or whether secondary hypogonadism precedes cognitive dysfunction in men at risk for AD. If this is true, testosterone replacement therapy for hypogonadal men at risk for dementia may be indicated. PMID- 14523359 TI - The effects of photoperiod on testes in rat: a morphometric and immunohistochemical study. AB - OBJECTIVE: The aim of this study was to examine the effects of photoperiod on testes in rat. DESIGN: For this purpose 21 male Wistar rats were used. Animals were divided into three groups. Control rats in group I were kept under 12 hrs light: 12 hrs dark conditions (12L: 12D) for 10 weeks. Animals in group II were exposed to long photoperiods (18L: 6D), while rats in group III were exposed to short photoperiods (6L: 18D) for 10 weeks. At the end of the experiment, all animals were killed by decapitation and blood samples were obtained. Serum testosterone levels were determined with the use of a chemiluminescent enzyme immunoassay. The testes of all rats were removed and weighed. Testicular tissue was processed semiquantitative evaluation of immunohistochemical testosterone staining. Intensity of immunostaining was determined on a scale between 0 (no staining) and 5 (heavy staining). For morphometric comparison, diameters of seminiferous tubules in each group were measured. RESULTS: In rats exposed to long photoperiods, testicular weights, diameters of seminiferous tubules and serum testosterone levels were significantly increased as compared to those in control rats. Whereas, exposure of rats to short photoperiods resulted in significantly decrease of testicular weights, diameters of seminiferous tubules and serum testosterone levels as compared to those in control rats and rats maintained in long photoperiods. Immunostaining of testosterone was moderate (3+) in control rats, heavy (5+) in rats exposed to long photoperiods and minimal (1+) in rats exposed to short photoperiods. CONCLUSIONS: The results of our study indicate that testicular functions increase after exposure to long photoperiods and decrease after exposure to short photoperiods. PMID- 14523360 TI - Melatonin supplementation restores cellular proliferation and DNA synthesis in the splenic and thymic lymphocytes of old rats. AB - OBJECTIVES: In this study we investigated the effect of melatonin treatment on the proliferative activity, the rate of DNA synthesis and the histopathological changes of splenic and thymic lymphocytes in old rats. METHODS: Two subgroups of old rats (25-months-old) were used in this study. One subgroup was given melatonin in the drinking water (250-300 mg/day/rat) for 3 months while the second subgroup was given water containing diluent. A third group consisted of young rats (3-months-old) which served as an additional control. RESULTS: A (3)H thymidine autoradiographic investigation showed a reduction in both the proliferative activity and the rate of DNA synthesis in splenic and thymic lymphocytes in old rats. In addition, light and electron microscopy showed severe histopathological changes in these cells from diluent-treated old rats. Melatonin administration increased the proliferative activity and the rate of DNA synthesis in the lymphocytes of both the spleen and thymus of the old animals. Also, histopathological changes were partially reversed by melatonin treatment with the tissues appearing similar to those in the young rats. CONCLUSION: The stimulation of the lymphocyte activity by melatonin is a beneficial response, especially in old rats, since aging results in an inhibition in lymphocytic functions. PMID- 14523361 TI - Direct effects of cocaine-amphetamine-regulated transcript (CART) on pituitary hormone release in pituitary cell culture. AB - OBJECTIVE: Cocaine- and amphetamine-regulated transcript (CART) is widely expressed in the rat brain, especially in the hypothalamic nuclei and in the anterior pituitary. The aim of this study was to evaluate the effects of CART on pituitary hormone release in pituitary cell culture. MATERIAL AND METHODS: The pituitary hormone release from pituitary cell culture after CART administration was investigated. Concentrations of LH, FSH, PRL, TSH and GH were measured with RIA methods. RESULTS: CART in all doses (1 nMol, 10 nMol, 100 nMol) stimulated prolactin (PRL) release and inhibited TSH release. CART administration caused a dose dependent decrease in LH release. CART did not change GH release from cultured pituitary cells. CONCLUSION: CART may affect directly pituitary hormones release in the cell culture. PMID- 14523362 TI - Chromacin-like peptide in leeches. AB - We demonstrate the presence in leech hemolymph of high levels of a peptide recognized by antiserum directed against bovine chromacin. The purification of the chromacin-like peptide was carried out by an acidic extraction, followed by solid phase and high pressure gel permeation chromatography and reversed-phase HPLC purification. Its sequence (GDFELPSIADPQATFESQRGPSAQQVDK) was established by a combination of techniques, including automated Edman degradation, MALDI-TOF measurement and DOT immunobinding assays with anti-chromogranin A. Mass spectrometry measurement revealed a m/z 3177Da, revealing the fact that the molecule is phosphorylated. ELISA titrations performed at each step of the purification revealed a major increase in the level of the peptide (ca. 125 nmol/microl of coelomic fluid) 15 min after LPS exposure. The increase in chromacin-like peptide levels is both time and concentration dependent. The level of this peptide decreased significantly 4 hours after LPS exposure. This report is the first discovery of a chromogranin derived like peptide in invertebrates. PMID- 14523363 TI - Peptides of pineal gland and thymus prolong human life. AB - OBJECTIVES AND DESIGN: Researchers of the St. Petersburg Institute of Bioregulation and Gerontology of the North-Western Branch of the Russian Academy of Medical Sciences and the Institute of Gerontology of the Ukrainian Academy of Medical Sciences (Kiev) clinically assessed the geroprotective effects of thymic (Thymalin) and pineal (Epithalamin) peptide bioregulators in 266 elderly and older persons during 6-8 years. The bioregulators were applied for the first 2-3 years of observation. RESULTS: The obtained results convincingly showed the ability of the bioregulators to normalize the basic functions of the human organism, i.e. to improve the indices of cardiovascular, endocrine, immune and nervous systems, homeostasis and metabolism. Homeostasis restoration was accompanied by a 2.0-2.4-fold decrease in acute respiratory disease incidence, reduced incidence of the clinical manifestations of ischemic heart disease, hypertension disease, deforming osteoarthrosis and osteoporosis as compared to the control. Such a significant improvement in the health state of the peptide treated patients correlated with decreased mortality rate during observation: 2.0 2.1-fold in the Thymalin-treated group; 1.6-1.8-fold in the Epithalamin-treated group; 2.5-fold in the patients treated with Thymalin plus Epithalamin as compared to the control. A separate group of patients was treated with Thymalin in combination with Epithalamin annually for 6 years and their mortality rate decreased 4.1 times as compared to the control. CONCLUSIONS: The obtained data confirmed the high geroprotective efficacy of Thymalin and Epithalamin and the expediency of their application in medicine and social care for health maintenance and age-related pathology prevention in persons over 60 to prolong their active longevity. PMID- 14523364 TI - Neonatal hypothalamic androgenization in the female rat induces changes in peripheral insulin sensitivity and adiposity function at adulthood. AB - It is recognized that there exists a link between hyperandrogenicity and insulin resistance. OBJECTIVE: By using the neonatally androgenized female rat we explored whether this treatment modifies peripheral insulin sensitivity and visceral fat function at adulthood. EXPERIMENTAL DESIGNS: On day 5 of age, female Sprague-Dawley pups were injected, sub cutaneous, with either 50 ml of sterile corn oil alone (CT) or containing 1.25 mg of testosterone propionate (TP) and further used for experimentation on day 100 of age. CT and TP rats were killed by decapitation in non-fasting condition and blood samples were kept frozen for measurement of different metabolites. Immediately after sacrifice, freshly dissected visceral fat pads were used for isolation of adipocytes, these cells were then incubated with medium alone or containing different concentrations of insulin in order to determine leptin secreted into the medium. Additionally, in vivo metabolic responses to intravenous high glucose load were performed in, 24 hour-fasting, CT and TP rats. RESULTS: We found that neonatal androgenization induced adult animals displaying higher visceral adiposity mass, body weight and leptinemia than CT rats. No group differences were found in basal circulating levels of several hormones and metabolic parameters. The results of the high glucose load 90-min test indicated that TP and CT rats developed similar glycemia but this accounted because of an early significantly higher peak values of circulating insulin in TP than in CT rats, regardless of similar enhancement in circulating glucocorticoid concentrations in both groups. While high glucose load significantly increased, over the baseline, circulating leptin concentrations as early as 30 min post-glucose in CT rats, in TP animals, it significantly enhanced leptinemia only by the end of the test. Finally, results of in vitro incubations of isolated visceral adipocytes indicated that cells from androgenized rats spontaneously released more leptin than control cells, although they were less responsive than CT cells to insulin-induced leptin output. CONCLUSION: Our study strongly supports the hypothesis that development of insulin resistance seems to be dependent on early hyperandrogenicity. PMID- 14523365 TI - Evaluation of the pituitary-adrenal axis in hyperandrogenic women with polycystic ovary syndrome. AB - OBJECTIVES: Impaired adrenal function is common in patients with polycystic ovary syndrome (PCOS). Abnormal regulation of cytochrome P450 17 alpha is believed to cause the exaggerated 17-hydroxyprogesterone (17OHP) response to ACTH stimulation. The aim of the study was to evaluate cortisol and 17OHP response to low dose (1 microg) ACTH test and to compare it with the standard ACTH (250 microg) test in hyperandrogenic women with PCOS. DESIGN AND MEASUREMENTS: We studied 27 PCOS and 22 control women. All participants were examined for mutations of the CYP21 gene, Cortisol and 17OHP levels before, 30 and 60 minutes after the IV injection of 250 microg ACTH (SDT) and after 1 microg ACTH (LDT). Fasting serum levels of LH, FSH, testosterone, DHEAS were determined in all participants. RESULTS: Basal and ACTH stimulated Cortisol during the SDT (470+/ 138 nmol/L and 761+/-143, respectively) were significantly higher in PCOS vs. controls (232+/-124 and 670+/-130, respectively) (p<0.03, p<0.02, respectively). Basal 17OHP (6.1+/-2.1 nmol/L) and the peak response to SDT (14.2+/-3.6 nmol/L) were significantly higher in PCOS vs. controls (4.2+/-2.1, 10.9+/-3.0, respectively) (p<0.003, p<0.004, respectively). Abnormally elevated 17OHP response to SDT was detected in 6/27 PCOS women (22%). No statistically significant difference between the PCOS and control groups were noted during the LDT in both cortisol and 17OHP levels. CONCLUSIONS: These data suggest that the exaggerated 17-hydroxyprogesterone (17OHP) response to ACTH stimulation in PCOS is revealed by stimulation at a pharmacological dose (250 microg) but not by a physiological dose (1 microg). PMID- 14523367 TI - Total pineal endocrine substitution therapy (TPEST) as a new neuroendocrine palliative treatment of untreatable metastatic solid tumor patients: a phase II study. AB - OBJECTIVES: It is known since many years that the pineal gland plays an anticancer role, and melatonin (MLT), the most investigated pineal hormone, has been proven to exert antitumor activity. However, MLT would not be the only hormone responsible for the antitumor action of the pineal gland. In fact, recent advances in the pineal investigations have shown that pineal indoles other than MLT may also exert anticancer activity, namely the three main indoles, consisting of 5-methoxytriptamine (5-MTT), 5-methoxytryptophol (5-MTP) and 5-methoxy-indole acetic acid (5-MIA). Cancer progression has appeared to be associated with a concomitant decline in the pineal endocrine function. Therefore, the replacement of a complete pineal function in the advanced cancer patients would require the exogenous administration of the overall four pineal indoles. Several clinical studies have shown that MLT alone at pharmacological doses may induce a control of the neoplastic progression in about 30% of untreatable metastatic solid tumor patients. The present study was performed in an attempt to evaluate the therapeutic of a total pineal endocrine substitution therapy with its four indole hormones in cancer patients, for whom no other conventional therapy was available. METHODS: The study included 14 metastatic solid tumor patients, who had failed to respond to the conventional anticancer therapies. The pineal indoles were given orally according to a schedule elaborated in an attempt to reproduce their physiological circadian secretion during the daily photoperiod. MLT was given at 20 mg/day during the night, whereas the other indoles were given at 1 mg/day, by administering 5-MIA in the morning, 5-MTP at noon and 5-MTT in the afternoon. RESULTS: A disease-control was achieved in 9/14 (64%) patients, consisting of partial response (PR) in one patient and stable disease (SD) in the other 8 patients. The median time of disease-control (PR + SD) was 6 months (range: 4-10). CONCLUSIONS: This preliminary study shows that a total pineal endocrine replacement therapy by an exogenous administration of the overall four pineal indoles may induce a disease-control in about 60% of untreatable metastatic solid tumor patients. Then, these results would be clearly superior with respect to those described with MLT alone, by confirming in humans that MLT is not the only hormone responsible for the anticancer property of the pineal gland. Since Cartesius was the first author who suggested the fundamental role of the pineal in the connection between consciousness and biological life, this therapy could be defined as a Cartesian therapy. PMID- 14523366 TI - Gastric emptying in subclinical hyperthyroidism. AB - INTRODUCTION AND AIM: About 15% of hyperthyroid patients suffer from gastrointestinal symptoms such as diarrhea, constipation, nausea and early satiety the problem. Published reports on the topic of gastric emptying in hyperthyroidism are rare and discrepant in their results. PATIENTS AND METHODS: Thirteen patients (8 women, 5 male; aged 41.92+/-1.491 SE yr) with a recently established diagnosis of subclinical hyperthyroidism due to Graves-Basedow disease and as control group 10 age-matched healthy subject (7 women, 3 male; 39.6+/-2.918 SE yrs) participated in this study. Scintigraphic method (food labeled with 99mTc sulfur colloid) was used for measurement of gastric emptying. RESULTS: Gastric emptying time was similar in both subclinical hyperthyroid and control groups (p=0.413). DISCUSSION: There are few studies on the subject of gastric emptying in hyperthyroidism with either delayed emptying or with emptying time similar to controls. In the only one study in subclinical hyperthyroidism, gastric emptying was found to be delayed. We also found that gastric emptying was delayed in subclinical hyperthyroidism. On the basis of this and other studies, we may suggest that in subclinical hyperthyroidism, gastric emptying of solids were similar to that in age-matched healthy control subjects and gastrointestinal dysfunction is dependent on severity of the hyperthyroid state and might be a result of impaired neuro-hormonal regulation. Yet the data must be supported on larger groups. PMID- 14523368 TI - Hormonal function and proliferative activity of thymic cells in humans: immunocytochemical correlations. AB - OBJECTIVES: The immunocytochemical study of the localization of hormones in thymic cells has been performed to clarify possible correlations of their expression with proliferative activity of thymocytes. METHODS: We used commercial antibodies to serotonin, melatonin, somatostatin, glucagon, gastrin, beta endorphin and histamine, and ABP or BSP kits for visualization of reaction. Computer image analysis was used to find correlations between hormone production and proliferative activity of thymocytes. RESULTS: Different subpopulations of thymocytes are able to produce hormones: precursors of T-lymphocytes (CD4-CD8-) contain serotonin and melatonin, immature cortical cells (CD4+CD8+) produce only serotonin, mature medullar cells (CD4+CD8-) show immunoreactivity to serotonin, melatonin, beta-endorphin and histamine. The expression of serotonin, somatostatin and gastrin is localized in thymic epithelial cells, formatting Gassal's bodies. Proliferative activity of thymocytes depends from the expression of serotonin and somatostatin in thymic cells. CONCLUSION: The data received testify the expression of different hormones in human thymic cells and showing by this fact high endocrine activity of thymus. The presence of correlations between hormonal expression and cell proliferative activity could be considered as the bright illustration of important role of neuroimmunoendocrine mechanisms in the regulation of local thymic homeostasis. PMID- 14523370 TI - Cardiac deaths after a mass smallpox vaccination campaign--New York City, 1947. AB - During the first wave of the 2003 smallpox vaccination campaign, two ischemic cardiac deaths occurred in civilian vaccinees aged 55 and 57 years, and one occurred in a military vaccinee aged 55 years, 4-17 days after vaccination with the New York City Board of Health (NYCBOH) vaccinia strain. Whether these and 13 other recognized military and civilian nonfatal ischemic events among vaccinees were associated with smallpox vaccination is unclear. The same NYCBOH strain was used in 1947 to vaccinate approximately six million New York City (NYC) residents (80% of the population) during a 4-week period (April 4-May 2) after a smallpox outbreak. To determine whether smallpox vaccination increased the risk for cardiac death in 1947, the NYC Department of Health and Mental Hygiene (DOHMH) analyzed data from NYC death certificates during that period. This report summarizes the results of that analysis, which found no increases in cardiac, atherosclerotic, or all-cause deaths. The findings are consistent with a growing body of evidence suggesting that ischemic cardiac deaths observed after the 2003 campaign might have been unrelated to vaccine. PMID- 14523369 TI - Ultrastructure of pinealocytes in mice implanted with Colon 38 adenocarcinoma. AB - OBJECTIVES: Relationship between the pineal gland and neoplastic disease has been repeatedly shown in many both experimental and clinical studies. However, morphological studies of the pineal gland in animals with experimentally-induced tumors are rare. Therefore, we decided to investigate the ultrastructure of pinealocytes in mice with implanted Colon 38 adenocarcinoma. MATERIAL AND METHODS: Male adult B6D2F1 mice were used in this study. The animals were divided into two groups. Eight mice were subcutaneously implanted with Colon 38 cell suspension, whereas the other eight intact animals served as controls. Three weeks after tumor implantation four animals from each group were sacrificed by spinal cord dislocation at 12:00 h or 24:00 h. The pineal glands were removed and processed for electron microscopic studies. The cross-sectional areas of the pinealocyte and its nucleus, and relative volume of mitochondria, Golgi apparatus, lysosomes, granular endoplasmic reticulum, and lipid droplets, as well as the number of dense-core vesicles were estimated using a digital analyzer connected on-line to IBM-PC computer. Statistical analysis of the data was performed using Student's t-test and Snedecor F test. RESULTS: In the pineal glands of the tumor-bearing animals killed during the daytime diminished size of pinealocytes and their nuclei, decreased relative volume of granular endoplasmic reticulum, lysosomes, and lipid droplets as well as decreased number of dense core vesicles were observed. On the contrary, the relative volumes of mitochondria and Golgi apparatus were increased in these animals. In the tumor bearing animals killed at night, however, increased cross-sectional areas of pinealocytes, and decreased number of dense-core vesicles were observed. CONCLUSIONS: The results of our study suggest that the presence of the malignant tumor influences the morphology of pineal cells in mice. Considering the fact that different patterns of ultrastructural changes were demonstrated in pinealocytes in different tumor types in various species, it seems that the character of the ultrastructural changes observed in tumor-bearing animals depends on the animal species and tumor type. PMID- 14523371 TI - Follow-up of deaths among U.S. Postal Service workers potentially exposed to Bacillus anthracis--District of Columbia, 2001-2002. AB - In October 2001, two letters contaminated with Bacillus anthracis spores were processed by mechanical and manual methods at the U.S. Postal Service (USPS) Brentwood Mail Processing and Distribution Center in the District of Columbia. Four postal workers at the Brentwood facility became ill with what was diagnosed eventually as inhalational anthrax; two died. The facility was closed on October 21, and postexposure prophylaxis was recommended for approximately 2,500 workers and business visitors. Subsequent reports of deaths of facility workers prompted concern about whether mortality was unusually high among workers, perhaps related to the anthrax attacks. To evaluate the rates and causes of death among workers at the Brentwood facility during October 12, 2001-October 11, 2002, CDC, in collaboration with state and local health departments, analyzed death certificate data. In addition, these data were compared with aggregate mortality data from the five USPS facilities contaminated with B. anthracis during the fall 2001 anthrax attacks. This report summarizes the results of that analysis, which indicate that rates and causes of death among Brentwood workers during the 12 months after the anthrax attacks of 2001 were not different from rates and causes of deaths that occurred during the preceding 5 years. PMID- 14523372 TI - Recognition of illness associated with exposure to chemical agents--United States, 2003. AB - Since September 11, 2001, concern has increased about potential terrorist attacks involving the use of chemical agents. In addition, recent cases involving intentional or inadvertent contamination of food with chemicals have highlighted the need for health-care providers and public health officials to be alert for patients in their communities who have signs and symptoms consistent with chemical exposures. For example, in February 2003, a Michigan supermarket worker was charged with intentionally contaminating 200 lbs. of meat with a nicotine containing insecticide. Although intentional release of chemical agents might be an overt event (i.e., one whose nature reveals itself), such as release of a nerve agent in a subway or a large explosion of a chemical container, a chemical release might instead be a covert event (i.e., an unrecognized release in which the presence of ill persons might be the first sign of an exposure), such as deliberate contamination of food, water, or a consumer product. To increase the likelihood that health-care providers will recognize a chemical-release-related illness and that public health authorities will implement the appropriate emergency response and public health actions, CDC identified examples of chemical induced illness and created appropriate guidance for health-care providers and public health personnel. This report summarizes the epidemiologic clues and clinical signs or patterns of illness that might suggest covert release of a chemical agent. CDC is working to develop national surveillance capabilities for detecting chemical-release-related illnesses. PMID- 14523373 TI - West Nile virus activity--United States, September 25-October 1, 2003. AB - This report summarizes West Nile virus (WNV) surveillance data reported to CDC through ArboNET as of 3 a.m., Mountain Daylight Time, October 1, 2003. PMID- 14523374 TI - Problems in detecting mosaic DNA methylation in Angelman syndrome. PMID- 14523375 TI - Limb-girdle muscular dystrophy 2I: phenotypic variability within a large consanguineous Bedouin family associated with a novel FKRP mutation. AB - Limb-girdle muscular dystrophies (LGMDs) represent a group of diseases characterized mainly by muscle wasting of the upper and lower limbs, with a wide range of clinical severity. The clinical heterogeneity is paralleled by molecular heterogeneity; each of the 10 forms of autosomal-recessive LGMD recognized to date is caused by mutations in a distinct gene. In a large consanguineous Bedouin tribe living in northern Israel, 15 individuals affected by LGMD demonstrate an autosomal recessive pattern of inheritance. A genome-wide screen followed by fine mapping in this family revealed linkage to a region on chromosome 19 harboring the fukutin-related protein gene (FKRP), with a maximal LOD score of 4.8 for D19S902. FKRP, encoding a putative glycosyltransferase, has been implicated in causing congenital muscular dystrophy 1C (MDC1C), and has recently been shown to be mutated in LGMD2I. We identified a novel missense mutation in exon 4 of the FKRP gene in all the patients studied. Although all affected individuals were homozygous for the same mutation, a marked phenotypic variability was apparent within the family. This finding may suggest a role of modifier genes and environmental factors in LGMD2I. Moreover, the demonstration that an identical, novel mutation in the FKRP gene can cause a muscle disease of either a congenital onset or of a later onset within a single family provides clinical support to the molecular evidence, suggesting that MDC1C and LGMD2I are overlapping ends of one and the same entity. PMID- 14523376 TI - Frequency and parental origin of de novo APC mutations in familial adenomatous polyposis. AB - A predominance of de novo mutations in the paternal germ line has been reported for several disorders; however, in familial adenomatous polyposis (FAP), the parental origin of APC mutations has been scarcely analysed so far. Among 563 unrelated FAP families with known family history, we identified 58 patients with a suspected de novo mutation in the APC gene. A germline mutation was detected in 52 of them; in 38 patients, the mutation could be excluded in both parents. The five base pair deletion at codon 1309 (c.3927_3931delAAAGA) was over-represented in the group of patients with suspected de novo mutations (17/58=29%), when compared to the group of familial cases (26/505=5%); thus, the high frequency of this mutation is not due to a founder effect but rather due to de novo mutation events. Parental origin of de novo mutations could be traced in 16 families, including three families with large chromosomal deletions. Four mutations were of maternal and 12 of paternal origin, pointing to a moderate preponderance towards paternal origin. Sex-related differences of mutation types could be observed: large deletions and single-base substitutions were exclusively of paternal origin, whereas the small deletions were equally distributed (maternal/paternal ratio 4:4). PMID- 14523377 TI - SLC11A1 (formerly NRAMP1) and susceptibility to visceral leishmaniasis in The Sudan. AB - Genetic susceptibility to visceral leishmaniasis (VL) is indicated by differences in incidence and clinical phenotypes between ethnic groups in Sudan. In mice, innate susceptibility to Leishmania donovani, the etiological agent of VL, is controlled by Slc11a1 (formerly Nramp1). We therefore examined polymorphisms at SLC11A1 in 59 multicase families of VL from the high-incidence Masalit tribe in Sudan. Multipoint nonparametric analysis in ALLEGRO shows a significant linkage across SLC11A1 (Zlr scores 2.38-2.55; 0.008< or =P< or =0.012; information content 0.88). The extended transmission disequilibrium test shows biased transmission of alleles at 5' polymorphisms in the promoter (P=0.0145), exon 3 (P=0.0037) and intron 4 (P=0.0049), and haplotypes formed by them (P=0.0089), but not for 3' polymorphisms at exon 15 or the 3'UTR. Stepwise logistic regression analysis using a case/pseudo-control data set derived from the 59 families was consistent with main effects contributed by the intron 4 469+14G/C polymorphism. Although the two alleles for 469+14G/C lie on haplotypes carrying different alleles for the functional promoter GTn polymorphism, the latter did not itself contribute separate main effects. Sequence analysis of 36 individuals failed to identify new putative functional polymorphisms in the coding region, intron 1, intron/exon boundaries, intron 4/exon 4a, or in the 3'UTR. One novel promoter polymorphism (-86G/A) was located within a putative nuclear factor kappa B binding site that could be functional. Further work will determine whether additional polymorphisms occur upstream in the promoter, which could be in linkage disequilibrium with the intron 4 polymorphism. These studies contribute to knowledge of the role of SLC11A1 in infectious disease. PMID- 14523378 TI - Multiparametric corticofugal modulation and plasticity in the auditory system. PMID- 14523379 TI - Neural crest specification: migrating into genomics. PMID- 14523380 TI - Cranial neural crest and the building of the vertebrate head. PMID- 14523381 TI - Neurobiology of suicidal behaviour. PMID- 14523382 TI - Working memory: looking back and looking forward. PMID- 14523383 TI - Moral cognition and its neural constituents. PMID- 14523384 TI - From neural 'is' to moral 'ought': what are the moral implications of neuroscientific moral psychology? PMID- 14523385 TI - Roles of the NKG2D immunoreceptor and its ligands. AB - According to present concepts, innate immunity is regulated by receptors that determine danger levels by responding to molecules that are associated with infection or cellular distress. NKG2D is, perhaps, the best characterized receptor that is associated with responses to cellular distress, defined as transformation, infection or cell stress. This review summarizes recent findings that concern NKG2D, its ligands, its signalling properties and its role in disease, and provides a framework for considering how the induction of immune responses can be regulated by cellular responses to injury. PMID- 14523386 TI - B cells under influence: transformation of B cells by Epstein-Barr virus. AB - Epstein-Barr virus (EBV) is an extremely successful virus, infecting more than 90% of the human population worldwide. After primary infection, the virus persists for the life of the host, usually as a harmless passenger residing in B cells. However, EBV can transform B cells, which can result in the development of malignant lymphomas. Intriguingly, the three main types of EBV-associated B-cell lymphoma - that is, Burkitt lymphoma, Hodgkin lymphoma and post-transplant lymphomas - seem to derive from germinal-centre B cells or atypical survivors of the germinal-centre reaction in most, if not all, cases, indicating that EBV infected germinal-centre B cells are at particular risk for malignant transformation. PMID- 14523387 TI - The SAP and SLAM families in immune responses and X-linked lymphoproliferative disease. AB - SAP (signalling lymphocytic activation molecule (SLAM)-associated protein) is a T and natural killer (NK)-cell-specific protein containing a single SH2 domain encoded by a gene that is defective or absent in patients with X-linked lymphoproliferative syndrome (XLP). The SH2 domain of SAP binds with high affinity to the cytoplasmic tail of the haematopoietic cell-surface glycoprotein SLAM and five related receptors. SAP regulates signal transduction of the SLAM family receptors by recruiting SRC kinases. Similarly, the SAP-related proteins EAT2A and EAT2B are thought to control signal transduction that is initiated by SLAM-related receptors in professional antigen-presenting cells. In this review, we discuss recent findings on the structure and function of proteins of the SAP and SLAM families. PMID- 14523388 TI - Plasma-cell homing. AB - Recent studies indicate that chemoattractant cytokines (chemokines), together with tissue-specific adhesion molecules, coordinate the migration of antibody secreting cells (ASCs) from their sites of antigen-driven differentiation in lymphoid tissues to target effector tissues. Developing ASCs downregulate the expression of receptors for lymphoid tissue chemokines and selectively upregulate the expression of chemokine receptors that might target the migration of IgA ASCs to mucosal surfaces, IgG ASCs to sites of tissue inflammation and both types of ASC to the bone marrow - an important site for serum antibody production. By directing plasma-cell homing, chemokines might help to determine the character and efficiency of mucosal, inflammatory and systemic antibody responses. PMID- 14523389 TI - Individuality: the barrier to optimal immunosuppression. AB - Immunosuppressive therapy aims to protect transplanted organs from host responses. Individuals have unique repertoires of responses to foreign antigens and toxic reactions to immunosuppressants; the former determining the type or intensity of rejection reactions and the latter influencing the severity of iatrogenic effects. Because existing agents target molecules that are widely distributed in tissues, new strategies must selectively block lymphoid cells only, disrupt alloresponses but not innate immune responses, interact synergistically with other agents, facilitate the homeostatic process that naturally leads to graft acceptance and ideally only interrupt donor-specific responses. Approaches presently under investigation aim to alter cell trafficking, or selectively deviate the maturation of antigen-presenting cells or inhibit lymphocyte-activation cascades - events that are crucial to rejection responses. PMID- 14523390 TI - Lasker goes transcriptional. PMID- 14523392 TI - Diverse roles for ubiquitin-dependent proteolysis in transcriptional activation. AB - A growing literature points to a fundamental role for the ubiquitin-proteasome degradation system (UPS) in transcription. Four recent publications add significant insight to our understanding of the connections between these processes. Each provides evidence that some aspect of the UPS can stimulate the activity of transcriptional activators. UPS might promote transcription by several mechanisms, and in some cases, even the final step of the UPS - proteolysis - might enhance activator function. PMID- 14523393 TI - How attraction turns to repulsion. PMID- 14523394 TI - Hippo and its mission for growth control. PMID- 14523395 TI - From shoot to root. PMID- 14523396 TI - Cdc34: cycling on and off the SCF. PMID- 14523397 TI - Of fertility, cystic fibrosis and the bicarbonate ion. PMID- 14523398 TI - Timing the cell cycle. PMID- 14523399 TI - Wnt signalling gets XEEKy. PMID- 14523400 TI - Can Fizzy fly solo? PMID- 14523401 TI - Human Langerhans cell in a dermal lymphatic vessel. PMID- 14523402 TI - Stability and association of Smoothened, Costal2 and Fused with Cubitus interruptus are regulated by Hedgehog. AB - The mechanisms involved in transduction of the Hedgehog (Hh) signal are of considerable interest to developmental and cancer biologists. Stabilization of the integral membrane protein Smoothened (Smo) at the plasma membrane is a crucial step in Hh signalling but the molecular events immediately downstream of Smo remain to be elucidated. We have shown previously that the transcriptional mediator Cubitus interruptus (Ci) is associated in a protein complex with at least two other proteins, the kinesin-like Costal2 (Cos2) and the serine threonine kinase Fused (Fu). This protein complex governs the access of Ci to the nucleus. Here we show that, consequent on the stabilization of Smo, Cos2 and Fu are destabilized. Moreover, we find that the Cos2-Fu-Ci protein complex is associated with Smo in membrane fractions both in vitro and in vivo. We also show that Cos2 binding on Smo is necessary for the Hh-dependent dissociation of Ci from this complex. We propose that the association of the Cos2 protein complex with Smo at the plasma membrane controls the stability of the complex and allows Ci activation, eliciting its nuclear translocation. PMID- 14523403 TI - In praise of Gates. PMID- 14523404 TI - Mosquito production mooted as fast track to malaria vaccine. PMID- 14523405 TI - China plays cards close to its chest over manned space shot. PMID- 14523406 TI - NIH 'roadmap' charts course to tackle big research issues. PMID- 14523407 TI - Ion-powered probe set for year-long Moon trek. PMID- 14523408 TI - Harvard heralds fresh take on systems biology. PMID- 14523409 TI - Adviser gears up to bring scientists to matters of state. PMID- 14523410 TI - Research council plan for Europe gets up steam. PMID- 14523411 TI - Wellcome to fund publication in open-access journals. PMID- 14523412 TI - China takes centre stage for liver proteome. PMID- 14523413 TI - Planck makes permanent posts. PMID- 14523416 TI - Across the great divide. PMID- 14523417 TI - Outline for a European research council. PMID- 14523418 TI - How scientists can help to protect US homeland. PMID- 14523424 TI - Sedimentology: floods of record. PMID- 14523423 TI - Gene autonomy: positions, please... PMID- 14523425 TI - Immunology: dangerous liaisons. PMID- 14523426 TI - Developmental biology: twisting the body into shape. PMID- 14523427 TI - Chemistry: mirrors in flatland. PMID- 14523428 TI - Neurobiology: backchat at the synapse. PMID- 14523429 TI - Materials science: molecules squeezed and stroked. PMID- 14523430 TI - Developmental biology: partners united. PMID- 14523431 TI - Cell biology: the hippo hypothesis. PMID- 14523433 TI - Obituary: Patricia S. Goldman-Rakic (1937-2003). PMID- 14523435 TI - Animal welfare: captivity effects on wide-ranging carnivores. PMID- 14523436 TI - Explosives: a microsensor for trinitrotoluene vapour. PMID- 14523437 TI - Binary switches and modification cassettes in histone biology and beyond. AB - An immense number of post-translational modifications on histone proteins have been described and additional sites of modification are still being uncovered. Whereas many direct and indirect connections between certain histone modifications and distinct biological phenomena have now been established, concepts for comprehending the extreme density and variety of these covalent modifications are lacking. Here, we formally introduce localized 'binary switches' and 'modification cassettes' as new concepts in histone biology, elucidating mechanisms that might govern the biological readout of distinct modification patterns. Specifically, our hypotheses provide missing models for the dynamic readout of stable histone modifications and offer explanations for several long-standing questions embedded in the literature. Our ideas might also apply to non-histone proteins and are open to direct experimental examination. PMID- 14523438 TI - Evolution of complex life cycles in helminth parasites. AB - The fundamental question of how complex life cycles--where there is typically more than one host-evolve in host--parasite systems remains largely unexplored. We suggest that complex cycles in helminths without penetrative infective stages evolve by two essentially different processes, depending on where in the cycle a new host is inserted. In 'upward incorporation', a new definitive host, typically higher up a food web and which preys on the original definitive host, is added. Advantages to the parasite are avoidance of mortality due to the predator, greater body size at maturity and higher fecundity. The original host typically becomes an intermediate host, in which reproduction is suppressed. In 'downward incorporation', a new intermediate host is added at a lower trophic level; this reduces mortality and facilitates transmission to the original definitive host. These two processes should also apply in helminths with penetrative infective stages, although the mathematical conditions differ. PMID- 14523439 TI - Programmable computing with a single magnetoresistive element. AB - The development of transistor-based integrated circuits for modern computing is a story of great success. However, the proved concept for enhancing computational power by continuous miniaturization is approaching its fundamental limits. Alternative approaches consider logic elements that are reconfigurable at run time to overcome the rigid architecture of the present hardware systems. Implementation of parallel algorithms on such 'chameleon' processors has the potential to yield a dramatic increase of computational speed, competitive with that of supercomputers. Owing to their functional flexibility, 'chameleon' processors can be readily optimized with respect to any computer application. In conventional microprocessors, information must be transferred to a memory to prevent it from getting lost, because electrically processed information is volatile. Therefore the computational performance can be improved if the logic gate is additionally capable of storing the output. Here we describe a simple hardware concept for a programmable logic element that is based on a single magnetic random access memory (MRAM) cell. It combines the inherent advantage of a non-volatile output with flexible functionality which can be selected at run time to operate as an AND, OR, NAND or NOR gate. PMID- 14523440 TI - Controlling anisotropic nanoparticle growth through plasmon excitation. AB - Inorganic nanoparticles exhibit size-dependent properties that are of interest for applications ranging from biosensing and catalysis to optics and data storage. They are readily available in a wide variety of discrete compositions and sizes. Shape-selective synthesis strategies now also yield shapes other than nanospheres, such as anisotropic metal nanostructures with interesting optical properties. Here we demonstrate that the previously described photoinduced method for converting silver nanospheres into triangular silver nanocrystals--so-called nanoprisms--can be extended to synthesize relatively monodisperse nanoprisms with desired edge lengths in the 30-120 nm range. The particle growth process is controlled using dual-beam illumination of the nanoparticles, and appears to be driven by surface plasmon excitations. We find that, depending on the illumination wavelengths chosen, the plasmon excitations lead either to fusion of nanoprisms in an edge-selective manner or to the growth of the nanoprisms until they reach their light-controlled final size. PMID- 14523441 TI - Enantiospecific electrodeposition of a chiral catalyst. AB - Many biomolecules are chiral--they can exist in one of two enantiomeric forms that only differ in that their structures are mirror images of each other. Because only one enantiomer tends to be physiologically active while the other is inactive or even toxic, drug compounds are increasingly produced in an enantiomerically pure form using solution-phase homogeneous catalysts and enzymes. Chiral surfaces offer the possibility of developing heterogeneous enantioselective catalysts that can more readily be separated from the products and reused. In addition, such surfaces might serve as electrochemical sensors for chiral molecules. To date, chiral surfaces have been obtained by adsorbing chiral molecules or slicing single crystals so that they exhibit high-index faces, and some of these surfaces act as enantioselective heterogeneous catalysts. Here we show that chiral surfaces can also be produced through electrodeposition, a relatively simple solution-based process that resembles biomineralization in that organic molecules adsorbed on surfaces have profound effects on the morphology of the inorganic deposits. When electrodepositing a copper oxide film on an achiral gold surface in the presence of tartrate ion in the deposition solution, the chirality of the ion determines the chirality of the deposited film, which in turn determines the film's enantiospecificity during subsequent electrochemical oxidation reactions. PMID- 14523442 TI - Episodic sediment accumulation on Amazonian flood plains influenced by El Nino/Southern Oscillation. AB - Continental-scale rivers with a sandy bed sequester a significant proportion of their sediment load in flood plains. The spatial extent and depths of such deposits have been described, and flood-plain accumulation has been determined at decadal timescales, but it has not been possible to identify discrete events or to resolve deposition on near-annual timescales. Here we analyse (210)Pb activity profiles from sediment cores taken in the pristine Beni and Mamore river basins, which together comprise 720,000 km2 of the Amazon basin, to investigate sediment accumulation patterns in the Andean-Amazonian foreland. We find that in most locations, sediment stratigraphy is dominated by discrete packages of sediments of uniform age, which are typically 20-80 cm thick, with system-wide recurrence intervals of about 8 yr, indicating relatively rare episodic deposition events. Ocean temperature and stream flow records link these episodic events to rapidly rising floods associated with La Nina events, which debouch extraordinary volumes of sediments from the Andes. We conclude that transient processes driven by the El Nino/Southern Oscillation cycle control the formation of the Bolivian flood plains and modulate downstream delivery of sediments as well as associated carbon, nutrients and pollutants to the Amazon main stem. PMID- 14523443 TI - Boundary-layer mantle flow under the Dead Sea transform fault inferred from seismic anisotropy. AB - Lithospheric-scale transform faults play an important role in the dynamics of global plate motion. Near-surface deformation fields for such faults are relatively well documented by satellite geodesy, strain measurements and earthquake source studies, and deeper crustal structure has been imaged by seismic profiling. Relatively little is known, however, about deformation taking place in the subcrustal lithosphere--that is, the width and depth of the region associated with the deformation, the transition between deformed and undeformed lithosphere and the interaction between lithospheric and asthenospheric mantle flow at the plate boundary. Here we present evidence for a narrow, approximately 20-km-wide, subcrustal anisotropic zone of fault-parallel mineral alignment beneath the Dead Sea transform, obtained from an inversion of shear-wave splitting observations along a dense receiver profile. The geometry of this zone and the contrast between distinct anisotropic domains suggest subhorizontal mantle flow within a vertical boundary layer that extends through the entire lithosphere and accommodates the transform motion between the African and Arabian plates within this relatively narrow zone. PMID- 14523444 TI - The oldest articulated chondrichthyan from the Early Devonian period. AB - Chondrichthyans (including living sharks, skates, rays and chimaeras) have a fossil record of scales and dermal denticles perhaps dating back to the Late Ordovician period, about 455 million years ago. Their fossil tooth record extends to the earliest Devonian period, almost 418 million years ago, whereas the oldest known articulated shark remains date from the Early Devonian period, about 394 million years ago. Here we report the discovery of an articulated shark that is almost 409 million years old from the Early Devonian (early Emsian) period of New Brunswick, Canada. The specimen, identified as Doliodus problematicus (Woodward), sheds light on the earliest chondrichthyans and their interrelationships with basal jawed vertebrates. This species has been truly problematic. Previously known only from isolated teeth, it has been identified as an acanthodian and a chondrichthyan. This specimen is the oldest shark showing the tooth families in situ, and preserves one of the oldest chondrichthyan braincases. More notably, it shows the presence of paired pectoral fin-spines, previously unknown in cartilaginous fishes. PMID- 14523445 TI - Temperature excludes N2-fixing heterocystous cyanobacteria in the tropical oceans. AB - Whereas the non-heterocystous cyanobacteria Trichodesmium spp. are the dominant N2-fixing organisms in the tropical oceans, heterocystous species dominate N2 fixation in freshwater lakes and brackish environments such as the Baltic Sea. So far no satisfactory explanation for the absence of heterocystous cyanobacteria in the pelagic of the tropical oceans has been given, even though heterocysts would seem to represent an ideal strategy for protecting nitrogenase from being inactivated by O2, thereby enabling cyanobacteria to fix N2 and to perform photosynthesis simultaneously. Trichodesmium is capable of N2 fixation, apparently without needing to differentiate heterocysts. Here we show that differences in the temperature dependence of O2 flux, respiration and N2 fixation activity explain how Trichodesmium performs better than heterocystous species at higher temperatures. Our results also explain why Trichodesmium is not successful in temperate or cold seas. The absence of heterocystous cyanobacteria in the pelagic zone of temperate and cold seas, however, requires another explanation. PMID- 14523446 TI - Jelly belly protein activates the receptor tyrosine kinase Alk to specify visceral muscle pioneers. AB - The secreted protein Jelly belly (Jeb) is required for an essential signalling event in Drosophila muscle development. In the absence of functional Jeb, visceral muscle precursors are normally specified but fail to migrate and differentiate. The structure and distribution of Jeb protein implies that Jeb functions as a signal to organize the development of visceral muscles. Here we show that the Jeb receptor is the Drosophila homologue of anaplastic lymphoma kinase (Alk), a receptor tyrosine kinase of the insulin receptor superfamily. Human ALK was originally identified as a proto-oncogene, but its normal function in mammals is not known. In Drosophila, localized Jeb activates Alk and the downstream Ras/mitogen-activated protein kinase cascade to specify a select group of visceral muscle precursors as muscle-patterning pioneers. Jeb/Alk signalling induces the myoblast fusion gene dumbfounded (duf; also known as kirre) as well as org-1, a Drosophila homologue of mammalian TBX1, in these cells. PMID- 14523447 TI - Jeb signals through the Alk receptor tyrosine kinase to drive visceral muscle fusion. AB - The Drosophila melanogaster gene Anaplastic lymphoma kinase (Alk) is homologous to mammalian Alk, a member of the Alk/Ltk family of receptor tyrosine kinases (RTKs). We have previously shown that the Drosophila Alk RTK is crucial for visceral mesoderm development during early embryogenesis. Notably, observed Alk visceral mesoderm defects are highly reminiscent of the phenotype reported for the secreted molecule Jelly belly (Jeb). Here we show that Drosophila Alk is the receptor for Jeb in the developing visceral mesoderm, and that Jeb binding stimulates an Alk-driven, extracellular signal-regulated kinase-mediated signalling pathway, which results in the expression of the downstream gene duf (also known as kirre)--needed for muscle fusion. This new signal transduction pathway drives specification of the muscle founder cells, and the regulation of Duf expression by the Drosophila Alk RTK explains the visceral-mesoderm-specific muscle fusion defects observed in both Alk and jeb mutant animals. PMID- 14523448 TI - Differential regulation of EIN3 stability by glucose and ethylene signalling in plants. AB - Glucose is a global regulator of growth and metabolism that is evolutionarily conserved from unicellular microorganisms to multicellular animals and plants. In photosynthetic plants, glucose shows hormone-like activities and modulates many essential processes, including embryogenesis, germination, seedling development, vegetative growth, reproduction and senescence. Genetic and phenotypic analyses of Arabidopsis mutants with glucose-insensitive (gin) and glucose-oversensitive (glo) phenotypes have identified an unexpected antagonistic interaction between glucose and the plant stress hormone ethylene. The ethylene-insensitive etr1 and ein2 mutants have glo phenotypes, whereas the constitutive ethylene signalling mutant ctr1 is allelic to gin4 (refs 4, 5). The precise molecular mechanisms underlying the complex signalling network that governs plant growth and development in response to nutrients and plant hormones are mostly unknown. Here we show that glucose enhances the degradation of ETHYLENE-INSENSITIVE3 (EIN3), a key transcriptional regulator in ethylene signalling, through the plant glucose sensor hexokinase. Ethylene, by contrast, enhances the stability of EIN3. The ein3 mutant has a glo phenotype, and overexpression of EIN3 in transgenic Arabidopsis decreases glucose sensitivity. PMID- 14523449 TI - Role of visual pigment properties in rod and cone phototransduction. AB - Retinal rods and cones share a phototransduction pathway involving cyclic GMP. Cones are typically 100 times less photosensitive than rods and their response kinetics are several times faster, but the underlying mechanisms remain largely unknown. Almost all proteins involved in phototransduction have distinct rod and cone variants. Differences in properties between rod and cone pigments have been described, such as a 10-fold shorter lifetime of the meta-II state (active conformation) of cone pigment and its higher rate of spontaneous isomerization, but their contributions to the functional differences between rods and cones remain speculative. We have addressed this question by expressing human or salamander red cone pigment in Xenopus rods, and human rod pigment in Xenopus cones. Here we show that rod and cone pigments when present in the same cell produce light responses with identical amplification and kinetics, thereby ruling out any difference in their signalling properties. However, red cone pigment isomerizes spontaneously 10,000 times more frequently than rod pigment. This high spontaneous activity adapts the native cones even in darkness, making them less sensitive and kinetically faster than rods. Nevertheless, additional factors are probably involved in these differences. PMID- 14523450 TI - Haem can bind to and inhibit mammalian calcium-dependent Slo1 BK channels. AB - Haem is essential for living organisms, functioning as a crucial element in the redox-sensitive reaction centre in haemproteins. During the biogenesis of these proteins, the haem cofactor is typically incorporated enzymatically into the haem pockets of the apo-haemprotein as the functionally indispensable prosthetic group. A class of ion channel, the large-conductance calcium-dependent Slo1 BK channels, possesses a conserved haem-binding sequence motif. Here we present electrophysiological and structural evidence showing that haem directly regulates cloned human Slo1 channels and wild-type BK channels in rat brain. Both oxidized and reduced haem binds to the hSlo1 channel protein and profoundly inhibits transmembrane K+ currents by decreasing the frequency of channel opening. This direct regulation of the BK channel identifies a previously unknown role of haem as an acute signalling molecule. PMID- 14523453 TI - Physics temps. PMID- 14523451 TI - Staphylocoagulase is a prototype for the mechanism of cofactor-induced zymogen activation. AB - Many bacterial pathogens secrete proteins that activate host trypsinogen-like enzyme precursors, most notably the proenzymes of the blood coagulation and fibrinolysis systems. Staphylococcus aureus, an important human pathogen implicated in sepsis and endocarditis, secretes the cofactor staphylocoagulase, which activates prothrombin, without the usual proteolytic cleavages, to directly initiate blood clotting. Here we present the 2.2 A crystal structures of human alpha-thrombin and prethrombin-2 bound to a fully active staphylocoagulase variant. The cofactor consists of two domains, each with three-helix bundles; this is a novel fold that is distinct from known serine proteinase activators, particularly the streptococcal plasminogen activator streptokinase. The staphylocoagulase fold is conserved in other bacterial plasma-protein-binding factors and extracellular-matrix-binding factors. Kinetic studies confirm the importance of isoleucine 1 and valine 2 at the amino terminus of staphylocoagulase for zymogen activation. In addition to making contacts with the 148 loop and (pro)exosite I of prethrombin-2, staphylocoagulase inserts its N terminal peptide into the activation pocket of bound prethrombin-2, allosterically inducing functional catalytic machinery. These investigations demonstrate unambiguously the validity of the zymogen-activation mechanism known as 'molecular sexuality'. PMID- 14523454 TI - Going multidisciplinary. PMID- 14523456 TI - Reply to Hasle et al. PMID- 14523457 TI - GST polymorphisms and occurrence of second neoplasms after treatment of childhood leukemia. PMID- 14523459 TI - A potential role for PRDM12 in the pathogenesis of chronic myeloid leukaemia with derivative chromosome 9 deletion. PMID- 14523458 TI - Unrelated umbilical cord blood stem cell transplant after failure of haploidentical or matched unrelated donor hematopoietic stem cell transplant. PMID- 14523460 TI - Successful treatment of hyperuricemia with low doses of recombinant urate oxidase in four patients with hematologic malignancy and tumor lysis syndrome. PMID- 14523461 TI - Imatinib produces significantly superior molecular responses compared to interferon alfa plus cytarabine in patients with newly diagnosed chronic myeloid leukemia in chronic phase. AB - We analyzed molecular responses in 55 newly diagnosed chronic-phase chronic myeloid leukemia (CML) patients enrolled in a phase 3 study (the IRIS trial) comparing imatinib to interferon-alfa plus cytarabine (IFN+AraC). BCR-ABL/BCR% levels were measured by real-time quantitative RT-PCR and were significantly lower for the imatinib-treated patients at all time points up to 18 months, P<0.0001. The median levels for imatinib-treated patients continued to decrease and had not reached a plateau by 24 months. A total of 24 IFN+AraC-treated patients crossed over to imatinib. Once imatinib commenced, the median BCR ABL/BCR% levels in these patients were not significantly different to those on first-line imatinib for the equivalent number of months. The incidence of progression in imatinib-treated patients, defined by hematologic, cytogenetic or quantitative PCR criteria, was significantly higher in the patients who failed to achieve a 1 log reduction by 3 months or a 2 log reduction by 6 months, P=0.002. A total of 49 patients were screened for BCR-ABL kinase domain mutations. Mutations were detected in two imatinib-treated patients who crossed over from IFN+AraC and both lost their imatinib response. In conclusion, first-line imatinib-treated patients had profound reductions in BCR-ABL/BCR%, which significantly exceeded those of IFN+AraC-treated patients and early measurements were predictive of subsequent response. PMID- 14523462 TI - Dynamics of BCR-ABL mRNA expression in first-line therapy of chronic myelogenous leukemia patients with imatinib or interferon alpha/ara-C. AB - We sought to determine dynamics of BCR-ABL mRNA expression levels in 139 patients with chronic myelogenous leukemia (CML) in early chronic phase, randomized to receive imatinib (n=69) or interferon (IFN)/Ara-C (n=70). The response was sequentially monitored by cytogenetics from bone marrow metaphases (n=803) and qualitative and quantitative RT-PCR from peripheral blood samples (n=1117). Complete cytogenetic response (CCR) was achieved in 60 (imatinib, 87%) vs 10 patients (IFN/Ara-C, 14%) after a median observation time of 24 months. Within the first year after CCR, best median ratio BCR-ABL/ABL was 0.087%, (imatinib, n=48) vs 0.27% (IFN/Ara-C, n=9, P=0.025). BCR-ABL was undetectable in 25 cases by real-time PCR, but in only four patients by nested PCR. Median best response in patients with relapse after CCR was 0.24% (n=3) as compared to 0.029% in patients with continuous remission (n=52, P=0.029). We conclude that (i) treatment with imatinib in newly diagnosed CML patients is associated with a rapid decrease of BCR-ABL transcript levels; (ii) nested PCR may reveal residual BCR-ABL transcripts in samples that are negative by real-time PCR; (iii) BCR-ABL transcript levels parallel cytogenetic response, and (iv) imatinib is superior to IFN/Ara-C in terms of the speed and degree of molecular responses, but residual disease is rarely eliminated. PMID- 14523464 TI - Unsolved issues in CLL biology and management. PMID- 14523463 TI - EBV-related lymphoproliferative disorders in immunocompetent patients. PMID- 14523465 TI - High incidence of trisomies 1q, 9q, and 11q in multiple myeloma: results from a comprehensive molecular cytogenetic analysis. PMID- 14523466 TI - CLL B cells with a bimodal CD38 expression pattern: persistence of bimodal populations despite effective therapy for B-CLL. PMID- 14523467 TI - DNA sequencing-based HLA typing detects a B-cell ALL blast-specific mutation in HLA-A(*)2402 resulting in loss of HLA allele expression. PMID- 14523468 TI - Clonally expanded T cells in hairy cell leukemia patients are not leukemia specific. PMID- 14523469 TI - ZAP-70 expression is a prognostic factor in chronic lymphocytic leukemia. AB - B-cell chronic lymphocytic leukemia (B-CLL) is a heterogenous disease with a highly variable clinical course. Recent studies have shown that expression of the protein tyrosine kinase ZAP-70 may serve as a prognostic marker in B-CLL. Employing a semiquantitative RT-PCR assay, we examined purified leukemia B cells of 39 CLL patients for the expression of ZAP-70 mRNA transcripts. Significant ZAP 70 mRNA levels exceeding those found in control samples with 5% T cells were detected in 36% of the CLL cases. Patients in the ZAP-70 positive cohort were characterized by an unfavorable clinical course with a significantly shorter progression-free survival as compared to the ZAP-70-negative patients (64%). These results were confirmed by flow-cytometric analysis of the ZAP-70 protein, and expanded to a larger patient cohort (n=67). A combined statistical analysis of 79 patients showed that the two patient subgroups also differed with regard to overall survival and a panel of known clinical prognostic factors including LDH, thymidine kinase serum levels and expression of the CD38 surface antigen by the leukemic cell clone. The level of ZAP-70 expression did not change over time in the majority of patients where sequential samples were available for analysis. PMID- 14523470 TI - Somatic mitochondrial DNA mutations in adult-onset leukaemia. AB - Mitochondrial genome instability has recently been demonstrated in a wide variety of human tumours and is implicated in the development of the myelodysplastic syndromes, a heterogeneous group of haematological disorders with an increased risk of malignant transformation. We therefore investigated the incidence of somatic mitochondrial DNA (mtDNA) mutations in patients with adult-onset leukaemia. We sequenced the entire mitochondrial genome from both normal tissue (buccal epithelial cells) and the leukaemia from 24 patients with adult-onset leukaemia. Somatic mtDNA mutation was present in nine individuals ( approximately 40%) and in each case the tumour genome differed from the normal genome sequence by a single sequence change. Using PCR-RFLP analysis and real-time PCR, we have studied in detail the mutation present in one patient with acute lymphatic leukaemia, demonstrating that the mutation is associated specifically with the leukaemia. PMID- 14523471 TI - Apoptosis induction in peripheral leukemia cells by remission induction treatment in vivo: selective depletion and apoptosis in a CD34+ subpopulation of leukemia cells. AB - In vitro studies demonstrating the induction of programmed cell death by cytotoxic drugs used in anticancer chemotherapy suggested that antileukemic treatment eliminates leukemia cells by apoptosis. We therefore analyzed apoptosis induction and activation of apoptosis signaling molecules in patients receiving remission induction treatment for AML and ALL during the initial phase of leukemia cell reduction. A coexistence of distinct populations of CD34(+) and CD34(-) leukemia cells could be identified. During chemotherapy, CD34(+) leukemia cells were more rapidly depleted than CD34(-) cells. Furthermore, a significant increase in leukemia cell apoptosis ex vivo was detected in CD34(+) cells, while no such increase was observed in the CD34(-) subpopulation, suggesting that CD34(+) leukemia cells are the main targets for apoptosis induction through antileukemic treatment. No alterations in Bax and Bcl-2 expression were found during in vivo chemotherapy, and CD95 expression and sensitivity remained low, indicating the induction of apoptosis independent of the CD95 system or regulation of protein levels of Bax and Bcl-2. The data suggest that analysis of leukemia cell subpopulations is required for further identification of apoptosis signaling molecules relevant for response to treatment and assessment of drug efficacy in vivo and in vitro. PMID- 14523472 TI - Lack of asparagine depletion in the cerebrospinal fluid after one intravenous dose of PEG-asparaginase: a window study at initial diagnosis of childhood ALL. PMID- 14523473 TI - Centrosome aberrations as a possible mechanism for chromosomal instability in non Hodgkin's lymphoma. AB - Recently, centrosome aberrations have been described as a possible cause of aneuploidy in many solid tumors. To investigate whether centrosome aberrations occur in non-Hodgkin's lymphoma (NHL) and correlate with histologic subtype, karyotype, and other biological disease features, we examined 24 follicular lymphomas (FL), 18 diffuse large-B-cell lymphomas (DLCL), 33 mantle cell lymphomas (MCL), and 17 extranodal marginal zone B-cell lymphomas (MZBCL), using antibodies to centrosomal proteins. All 92 NHL displayed numerical and structural centrosome aberrations as compared to nonmalignant lymphoid tissue. Centrosome abnormalities were detectable in 32.3% of the cells in NHL, but in only 5.5% of lymphoid cells from 30 control individuals (P<0.0001). Indolent FL and MZBCL contained only 25.8 and 28.8% cells with abnormal centrosomes. In contrast, aggressive DLCL and MCL harbored centrosome aberrations in 41.8 and 35.0% of the cells, respectively (P<0.0001). Centrosomal aberrations correlated to lymphoma grade, mitotic, and proliferation indices, but not to the p53 labeling index. Importantly, diploid MCL contained 31.2% cells with abnormal centrosomes, while tetraploid samples harbored centrosome aberrations in 55.6% of the cells (P<0.0001). These results indicate that centrosome defects are common in NHL and suggest that they may contribute to the acquisition of chromosomal instability typically seen in NHL. PMID- 14523474 TI - Cobalt chloride and low oxygen tension trigger differentiation of acute myeloid leukemic cells: possible mediation of hypoxia-inducible factor-1alpha. AB - Cellular and systemic O(2) concentrations are tightly regulated to maintain delicate oxygen homeostasis. Although the roles of hypoxia in solid tumors have been widely studied, few studies were reported regarding the possible effects of hypoxia on leukemic cells. Here, we showed for the first time that low concentrations of cobalt chloride (CoCl(2)), a hypoxia-mimicking agent, and 2-3% O(2) triggered differentiation of various subtypes of human acute myeloid leukemic (AML) cell lines, including NB4, U937 and Kasumi-1 cells, respectively, from M3, M5 and M2b-type AML, but CoCl(2) did not modulate AML subtype-specific fusion proteins promyelocytic leukemia-retinoic acid receptor alpha (PML RARalpha) and AML1-ETO. Treatment with CoCl(2) also induced primary leukemic cells from some AML patients to undergo differentiation. Similar to what occurs in solid tumor cells, CoCl(2)-mimicked hypoxia also increased the level of hypoxia-inducible factor (HIF)-1alpha protein and its DNA-binding activity in leukemic cells. The CoCl(2) induction of HIF-1alpha protein and its DNA-binding activity were inhibited by 3-morpholinosydnonimine, which also blocked CoCl(2) induced cell differentiation in leukemic cells. These results provide an insight into a possible link of hypoxia or HIF-1alpha and leukemic cell differentiation, and are possibly of significance to explore clinical potentials of hypoxia or hypoxia-mimicking agents and novel target-based drugs for differentiation therapy of leukemia. PMID- 14523475 TI - Amplification of AML1 in acute lymphoblastic leukemia is associated with a poor outcome. PMID- 14523476 TI - Flavopiridol downregulates the expression of both the inducible NO synthase and p27(kip1) in malignant cells from B-cell chronic lymphocytic leukemia. AB - Flavopiridol, an inhibitor of cyclin-dependent kinases and other protein kinases, induces in vitro apoptosis of malignant cells from B-cell chronic lymphocytic leukemia (B-CLL). Previously, we reported that nitric oxide (NO), produced by an inducible NO synthase (iNOS), spontaneously expressed by the B-CLL cells, contributed to their deficiency in apoptosis. In the present work, we show that ex vivo treatment of leukemic cells from B-CLL patients with flavopiridol results in the inhibition of iNOS expression, as determined by immunofluorescence and Western blotting, and in a marked inhibition of NO production measured in situ with a specific fluorescent probe (DAF-2 DA). These effects are accompanied by membrane, mitochondrial and nuclear events of apoptosis. Flavopiridol exposure also results in the stimulation of caspase 3 activity and in caspase-dependent cleavage of p27(kip1), a negative regulator of the cell cycle, which is overexpressed in B-CLL. Thus, flavopiridol is capable of downregulating both iNOS and p27(kip1) expression in B-CLL cells. Furthermore, flavopiridol-promoted apoptosis is partly reverted by an NO donor, suggesting that inhibition of the NO pathway could participate in the apoptotic effects of flavopiridol on the leukemic cells. PMID- 14523477 TI - Clinical significance of central nervous system involvement at diagnosis of pediatric acute myeloid leukemia: a single institution's experience. AB - To determine the clinical significance of central nervous system (CNS) involvement at the time of diagnosis of pediatric acute myeloid leukemia (AML), we analyzed clinical features and outcomes of 290 patients treated consecutively on four institutional trials (AML80, AML83, AML87, and AML91). CNS status was classified as CNS1 (no blast cells in CSF; n=205), CNS2 (<5 WBC/mul CSF with blast cells; n=37), or CNS3 (>/=5 WBC/mul CSF with blast cells, or signs of CNS involvement; n=48). Patients with CNS3 status were significantly younger than others (P=0.016) and significantly more likely to have the favorable cytogenetic features t(9;11), t(8;21), or inv(16) (P<0.001). The CNS3 group had a significantly greater probability (+/-s.e.) of 5-year event-free survival (43.7+/ 7.0%) than did the CNS1 (27.8+/-3.2%, P=0.015) and CNS2 (24.3+/-7.5%, P=0.032) groups. However, after adjustment for favorable genetic features, there was no significant difference in EFS between the CNS3 and the combined CNS1+CNS2 groups (P=0.075). In all, 10 of 151 patients treated on AML80 and AML83, but none of 139 treated on AML87 and AML91, had primary CNS relapse. CNS involvement had no adverse prognostic significance, and patients with CNS2 status had similar outcome to CNS1 patients in this large group of pediatric patients with AML, treated at a single institution. PMID- 14523478 TI - Autocrine lymphotoxin production in Epstein-Barr virus-immortalized B cells: induction via NF-kappaB activation mediated by EBV-derived latent membrane protein 1. AB - Epstein-Barr virus (EBV)-immortalized lymphoblastoid cells express high levels of lymphotoxin and use this molecule as an autocrine growth factor. We hypothesized that the EBV-derived latent membrane protein 1 (LMP1) mediates lymphotoxin production by inducing NF-kappaB binding to the lymphotoxin promoter. We assessed lymphotoxin production, LMP1 expression, and NF-kappaB activation in Z-43 (EBV positive lymphoblastoid cells), Daudi (EBV-positive Burkitt's cells), and 3A4 (EBV-negative Burkitt's cells containing a stably transfected tetracycline inducible LMP1 construct). Z-43 cells expressed high levels of LMP1 (immunoblot) and lymphotoxin (ELISA); the EBV-positive Burkitt's lymphoma line Daudi expressed neither LMP1 nor lymphotoxin. Similarly, induction of LMP1 in the 3A4 cells (exposed to tetracycline) was accompanied by a 13-fold increase in lymphotoxin levels (ELISA) as compared to uninduced (LMP1-negative) cells. EMSAs demonstrated high levels of NF-kappaB activation in Z-43 and tetracycline-induced 3A4 cells, but much lower levels in the uninduced 3A4 cells. Exposure of these cells to Bay 11-7082 (an inhibitor of IkappaB phosphorylation and, therefore, NF-kappaB activation) abrogated NF-kappaB binding and lymphotoxin production in a dose dependent manner in both Z-43 and 3A4 cells. Therefore, in our model system, autocrine lymphotoxin production is largely driven by NF-kappaB activation, which is in turn mediated by EBV-derived LMP1 signaling. PMID- 14523479 TI - Segmental chromosomal aberrations and centrosome amplifications: pathogenetic mechanisms in Hodgkin and Reed-Sternberg cells of classical Hodgkin's lymphoma? AB - Tumor cell metaphases of classical Hodgkin's lymphoma (cHL) characteristically display highly rearranged karyotypes with chromosome numbers in the hyperploid range and marked intraclonal variability. The causes of this cytogenetic pattern remain largely unknown. An unusual type of chromosomal abnormality coined as segmental chromosomal aberration (SCA) has been recurrently observed in HL cell lines and was suggested to be associated with ribosomal DNA (rDNA) rearrangements. Moreover, centrosome abnormalities provoking deficient chromosome segregation have been reported in many solid tumors and also in cHL cell lines. Whether SCA, rDNA rearrangements or centrosome abnormalities also occur in primary cHL is not yet known. Thus, we performed extensive molecular cytogenetic and immunohistological studies in two cHL cases. Both cases presented SCA associated with genomic gains of the REL and JAK2 loci, respectively. The SCA involving JAK2 was associated with rDNA rearrangements. The absolute centrosome size of HRS cells in both cases was significantly larger than in non-HRS cells, but the relative centrosome size of HRS cells corrected for nuclear size was in the same range as that of the non-neoplastic cells. These findings demonstrate that the various mechanisms associated with chromosomal instability warrant a more detailed characterization in cHL. PMID- 14523480 TI - BCL10 mutations are irrelevant to its aberrant nuclear localization in nasal NK/T cell lymphoma. PMID- 14523481 TI - Signal averaged ECG in different patterns of left ventricular hypertrophy and geometry in hypertension. AB - BACKGROUND: It is still unknown which factors determine the presence of ventricular late potentials (LP) in hypertension. AIM: To evaluate the prevalence of LP in hypertension in relation to the pattern of left ventricular hypertrophy (LVH) and geometry, and to establish the factors causing signal-averaged ECG abnormalities. METHODS: The study group consisted of 109 patients (58 females, 51 males, mean age 49.7 +/-9.1 years) with hypertension and without coronary artery disease. Two-dimensional Echo Doppler, 24-hr ECG Holter, signal-averaged ECG and spectral analysis of heart rate variability (HRV) were performed. Four patterns of LVH and geometry were identified: normal geometry (N; n=30), concentric remodelling (CR; n=24), concentric hypertrophy (CH; n=38) and eccentric hypertrophy (EH; n=17). RESULTS: LP were more frequently detected in patients with LVH (9.1%), particularly in those with EH, than in patients without LVH (5.6%). Linear regression analysis revealed no correlation between signal averaged ECG parameters and LV ejection fraction (LVEF) or diastolic LV function indices. None of echocardiographic variables correlated with signalaveraged QRS duration, however, a significant positive correlation between LAS and LV mass (LVM) (r=0.26), LAS and LV end-diastolic volume (EDV) (r=0.2), as well as a significant negative correlation between V40 and LVM (r=-0.22) were noted. A significant positive correlation between LF/HF and signal-averaged QRS (r=0.31) and LAS (r=0.29) as well as a significant negative correlation between LF/HF and V40 (r=-0.21) were found. In the univariate analysis, the presence of EH was significantly related to the occurrence of LP (p<0.01). The reduction of HF power <113 ms(2), indicating a withdrawal of parasympathetic activity, was associated with LP (p<0.05). A ratio of LF 1n/HF 1n >1.28, indicating relative sympathetic overactivity, was a relative risk for LP incidence (p<0.05). In the multivariate analysis, however, all these factors were not independent predictors of the presence of LP. CONCLUSIONS: LP are more frequently detected in hypertensives with LVH, particularly in those with eccentric hypertrophy pattern. Left ventricular structural remodelling and withdrawal of parasympathetic tone are the significant determinants of LP occurrence. PMID- 14523482 TI - Is the decrease of cardiovascular mortality in Poland associated with the reduction of global cardiovascular risk related to changes in life style? AB - BACKGROUND: After an ascending trend of cardiovascular mortality, which was observed up to the sixties, there was a declining tendency in western countries thereafter. The decrease in mortality rates in Poland has been noticed since 1991. There is an uncertainty whether this improvement in prognosis was only due to the improvement in health care level or was accomplished, to some extent, by population life style changes. AIM: To assess whether the decrease in cardiovascular mortality in Poland was accompanied by a simultaneous reduction in global cardiovascular risk profile. METHODS: The study was based on screened random samples from the Warsaw population aged 35-64 years, who were examined in the years 1984 (2646 subjects), 1988 (1433 subjects), 1993 (1539 subjects) and 2001 (853 subjects). The group evaluated in 1984 was followed up for 10 years with all fatal events recorded (364 deaths were registered including 166 cardiovascular deaths). The model of 10 years probability of total and cardiovascular death was developed, which was based on 11 risk factors (age, smoking cigarettes, systolic blood pressure, pulse pressure, ratio of total cholesterol/HDL-cholesterol, triglyceride, symptoms of coronary heart disease or heart failure, death of mother before 65 or death of father before 55 years due to myocardial infarction or stroke, energy in daily food intake and percentage of energy derived from saturated fatty acids). This model was applied to assess the probability of deaths between years 1984 and 2001, utilising data from screenings. RESULTS: The probability of death (in %) decreased from 1984 to 2001 by 11% in men. However, it remained stable for women. On the other hand the probability of cardiovascular death decreased by 25% and 33%, respectively. The main role in global risk decrease was played by beneficial trends in mean systolic blood pressure (in both genders) and percentage of smokers in men. CONCLUSIONS: Modification of life style influencing risk factors profile decreased the probability of death in the Warsaw population, correlating with changes in mortality rates. PMID- 14523483 TI - Efficacy and safety of percutaneous coronary interventions in patients with non ST segment elevation acute coronary syndrome in catheterisation laboratory without on-site surgical back-up. AB - BACKGROUND: Percutaneous coronary intervention (PCI) in patients with non ST segment elevation acute coronary syndrome (NSTEACS) is regarded as a procedure which carries a high risk of immediate and long-term adverse cardiac events. This may potentially limit the use of PCI in catheterisation laboratories which do not have on-site surgical back up. However, stents and GP IIb/IIIa receptor inhibitors improved safety of interventional procedures. AIM: To analyse the immediate and long-term outcome of patients with NSTEACS in whom PCI was performed in a catheterisation laboratory without on-site surgical back-up in. METHODS: In a cohort of 479 consecutive patients (160 with NSTEACS - group A, 319 with stable angina - group B) we analysed short and long-term clinical outcome of PCI performed in our catheterisation laboratory which is located several kilometres from a cardiac surgery department, with an effective transfer time <30 minutes. RESULTS: Stent implantation rate and the usage of GP IIb/IIIa blockers were higher in group A than in group B (61.3% vs 50.2%, p=0.04, and 17.5% vs 6.3%; p<0.001, respectively). The in- hospital outcome was similar in both groups (death: 0.6 vs 0.6%; myocardial infarction (MI): 2.5 vs 1.6%; and urgent reintervention (rePCI): 1.9 vs 1.3%, all differences NS). Acute PCI complications requiring urgent surgical operation occurred in 1 (0.6%) patient from group A and in 1 (0.3%) patient from group B (NS). Both patients were successfully transferred for cardiac surgery. During a long-term follow-up the incidence of death (2.0 vs 2.0%), MI (0.7 vs 0.7%), rePCI (21.8 vs 25.2%), CABG (1.4 vs 1.4%) or coronary rehospitalisation (5.4 vs 7.7%) was similar in both groups. The Kaplan-Meier survival and event-free curves were parallel. CONCLUSIONS: In the era of coronary stents and platelet GP IIb/IIIa receptor inhibitors the short and long-term outcome after PCI in patients with NESTACS and stable angina is similar. The early aggressive approach to patients with NESTACS is feasible and safe in a catheterisation laboratory without on-site cardiac surgery. Surgical back-up is still necessary for only few PCI complications. PMID- 14523484 TI - Acute myocardial infarction complicated by cardiogenic shock. In-hospital and mid term results of invasive treatment in the National Institute of Cardiology, Warsaw-Anin. AB - BACKGROUND: Mortality in acute myocardial infarction (MI) complicated by cardiogenic shock approaches 90%, regardless of the type of pharmacological treatment. AIM: To assess in-hospital and mid-term results of invasive treatment of patients with acute MI with ST segment elevation (STEMI) complicated by cardiogenic shock. METHODS: From a prospective registry of all patients admitted to our institution for urgent coronary angiography due to acute coronary syndrome between February 2001 and June 2002, patients with STEMI, symptom duration up to 12 hours and cardiogenic shock diagnosed on admission were identified. The in hospital and mid-term outcome of 37 patients (mean age 65 years, range 54-77, 68% of males) treated with primary percutaneous coronary intervention (PCI) was analysed. RESULTS: Of the 41 patients with STEMI and cardiogenic shock, total occlusion or critical stenosis of a coronary artery were found in 38 patients. One patient with the occlusion of three main coronary arteries underwent urgent surgical revascularisation and remains alive after an 18-month follow-up. In the remaining 37 patients primary PCI of an infarct-related artery was performed (stent implantation in 70%, abciximab administration in 54%) which restored normal blood flow (TIMI grade 3 flow) in 54% of subjects. In patients with TIMI grade 3 flow the in-hospital mortality was 25%. Of the whole PCI-treated group, 18 (48.6%) patients died during stay in our institution, an additional two - after transfer to another hospital, and one - during a 19-month follow-up period. The remaining 16 patients remain alive (median follow-up of 8 months). CONCLUSIONS: Invasive treatment of patients with STEMI complicated by cardiogenic shock significantly reduces mortality in this high-risk population. The mid-term results in patients discharged from hospital are good. Invasive treatment of acute MI should be accessible for all patients with extensive acute MI. PMID- 14523485 TI - Antioxidant vitamins decrease exercise-induced QT dispersion after myocardial infarction. AB - BACKGROUND: Antioxidant vitamins C and E inhibit neutrophil-mediated production of free radicals in acute myocardial infarction (MI) which may limit MI size and improve myocardial perfusion. AIM: To examine whether treatment with vitamin C and E reduces inhomogeneity of repolarisation in patients with acute MI. METHODS: In this double-blind, placebo-controlled randomised trial 37 patients with acute MI were enrolled and assigned to vitamin C and E (600 mg/day each) or placebo treatment, starting on the first day of acute MI and lasting for 14 days. Inhomogeneity of repolarisation was assessed by examining QT interval dispersion (QTd), measured both at rest and at the end of sub-maximal exercise test, performed before discharge. RESULTS: Baseline QTd was similar in both groups, however, exercise-induced QTd was significantly lower in patients treated with antioxidant vitamins compared with the placebo group (59 +/- 20 msec vs 74 +/- 24 msec, p<0.05). PMID- 14523486 TI - [Left ventricular thrombus in early phase after myocardial infarction. Risk and treatment methods based on an observed case]. AB - A 55-year-old man was admitted to our Department four weeks after anterior myocardial infarction. A large, mobile thrombus was diagnosed on echocardiography. On the third day of hospitalization systemic arterial embolism with left femoral artery localization occurred. The embolus was removed surgically. During 3-month follow-up the patient received anticoagulant therapy with complete resolution of ventricular thrombus. Therapeutic options in this case are discussed. PMID- 14523487 TI - [Coronary flow reserve in acute myocardial infarction in a patient treated with primary venous angioplasty. Case report]. AB - Residual stenosis, dissection and/or microvascular damage have been proposed as mechanisms of impaired flow after primary angioplasty (PCI) for acute myocardial infarction (MI). In this report we present a patient who underwent PCI for acute anterior MI. Coronary flow reserve (CFR) assessment performed after balloon dilatation suggested possible improvement of coronary flow after stenting. However, we did not observe any improvement in CFR just after stent implantation. We conclude that in patients treated with primary PCI the use of additional pharmacological treatment to prevent microcirculation injury during reperfusion and slow-flow or no-reflow phenomenon should be considered. PMID- 14523488 TI - [Intracranial hemorrhage complicated by recurrent polymorphic ventricular torsade de pointes tachycardia]. AB - We present a case of a 65-year-old female admitted to the hospital due to severe intracranial hemorrhagia complicated by recurrences of polymorphic ventricular tachycardia of torsade de pointes type. ECG showed a marked prolongation of QT interval and giant U waves. The potassium level was 2.9 mmol/l and magnesium level - 0.6 mmol/l. Intravenous lidocaine and magnesium caused a complete supression of arrhythmia. Unfortunately, the patient died three days later due to cerebral damage and respiratory failure. PMID- 14523489 TI - [Myocarditis: diagnostic problems]. PMID- 14523494 TI - [Electrocardiogram of the month]. PMID- 14523495 TI - [False diagnosis of Ebstein's anomaly]. PMID- 14523496 TI - [Angiogram of the month]. PMID- 14523497 TI - [BNP - new prognostic parameter in acute coronary syndromes?]. PMID- 14523499 TI - [Contributions of Polish cardiologists to Western literature]. PMID- 14523501 TI - [An update on posttraumatic stress disorder]. PMID- 14523502 TI - [Posttraumatic stress disorder: diagnostic criteria]. AB - This article reviews aspects of the clinical characterization of the Postttraumatic Stress Disorder (PTSD). The disgnostic criteria used in DSM-IV and ICD-10 are presented. The Acute Reaction to Stress is defined and contrasted to the operational criteria which define the PTSD. The importance of the partial expression of the PTSD syndrome in victims of trauma is highlighted. Systematic studies within this patient population is characterized as a unmet need in public health. Addressing such a need is a major step towards the proper diagnosis and treatment of this clinical syndrome. PMID- 14523504 TI - [Diagnosis of the posttraumatic stress disorder]. AB - Only recently, Brazilian psychiatry has acknowledged the importance of the diagnosis of posttraumatic stress disorder (PTSD). Despite the fact that Brazilians have a high prevalence of exposure to traumatic events, such as accidents and homicides, there are few theoretical and empirical studies about PTSD in Brazil. This paper provides a brief review of the diagnosis of PTSD. Following a discussion of the main issues on the definition of traumatic event, we focused our discussion on the challenges of the psychiatric examination of patients with PTSD. In the last section of the paper, the authors emphasize the description of the PTSD symptoms and provide clinical vignettes to illustrate important psychopathological concepts. In conclusion, the importance of the diagnosis of PTSD in psychiatry is highlighted inasmuch as it provides an observational framework for investigating the effects of stress and trauma. PMID- 14523503 TI - [Historical evolution of the concept of posttraumatic stress disorder]. AB - The authors elaborate on the historical evolution of the concept of Posttraumatic Stress Disorder (PTSD). The authors quote the French scholars, mainly Charcot and Janet, as the first to connect traumatic events and symptoms of hysteria. The contributions of Freud are described with enphasis on his effort into integrating the intra-psychic and environmental dimensions. Kardiner is referred as the author who coined the concept of 'war neurosis', which was deemed as an important one during the Second World War and Vietnam War. In conclusion, the authors highlight that the concept of PTSD used in the Diagnostic and Statistical Manual of Mental Disorders (DSM) by the American Psychiatric Association assess, at the same time, how treatening was the traumatic event and the list of symptoms presented by the patients. PMID- 14523505 TI - [Comorbidities in posttraumatic stress disorder: rule or exception?]. AB - Posttraumatic stress disorder (PTSD) commonly occurs in conjunction with other psychiatric disorders. The present article reviews PTSD and its comorbidities such as anxiety disorders, substance abuse, mood disorders, suicide attempts, dissociative disorder and somatoform disorder. PMID- 14523506 TI - [Biological basis of posttraumatic stress disorder]. AB - Neuroendocrinological research on the physiological systems involved in stress evidenced hyper functioning of the sympatho-adrenal axis together with reduced activity of the hypothalamic-pituitary-adrenal axis in patients with posttraumatic stress disorder (PTSD). An impaired corticoid response to stressors seems to be associated with enhanced vulnerability to PTSD. Excess catecholamines, unchecked by corticoids would promote over consolidation of traumatic memories and undue generalization to other stressful situations. Symptoms such as numbing and flashbacks have been related to endogenous opioids. Neuroimaging studies evidenced a reduction of hippocampal volume in PTSD patients, which has been related to both cognitive changes and abnormalities of the HPA axis that are found in PTSD. PMID- 14523507 TI - [Memory consolidation and posttraumatic stress disorder]. AB - Extensive evidence from animal and human studies has shown that memory formation is enhanced by an endogenous modulatory system mediated by stress hormones and activation of the amygdala. This system is an evolutionarily adaptive method of enhancing important memories. Under emotional stress, this system is activated promoting the formation of vivid, long lasting traumatic memories, which are the hallmark of PTSD. The understanding of the mechanisms underlying memory modulation might lead to an improved ability to assess and treat PTSD. PMID- 14523508 TI - [Cognitive therapy for posttraumatic stress disorder]. AB - The authors describe the clinical presentation of Posttraumatic Stress Disorder (PTSD) according to the cognitive-behavioral model. The vulnerability factors for the development of PTSD are discussed. The foundations of the cognitive approach to PTSD are presented and guidelines on the contends and duration of the psychotherapy is proposed. PMID- 14523509 TI - [Posttraumatic stress disorder: the psychoanalytic approach]. AB - The authors approach the posttraumatic stress disorder according to its importance within the epidemiologic studies and public health figures. A brief history on how the concepts of psychic trauma developed within psychoanalysis is presented. The importance given by Freud to traumatic events as a determinant of neurosis is revisited. Trauma is discussed as part of the internal world perspective and not as dependent upon the external events. The importance of the use of psychoanalytic techniques to treat some cases of PTSD is highlighted. PMID- 14523510 TI - [The role of psychological debriefing in the treatment of victims of trauma]. AB - The application of psychological debriefing has become an expected and widespread intervention following exposure to trauma. This article assesses the wisdom of such an approach and reports upon expert consensus regarding its use. Meta analytic and narrative reviews are summarised and areas of agreement and disagreement are outlined. In sum, it was concluded that the majority of people do not become traumatised from stressful events; that generic psychological debriefing, when applied to individuals, appears to have little impact on functioning; that a specific form of debriefing called Critical Incident Stress Debriefing holds the possibility of noxious effects for some participants and that those most deleteriously affected by debriefing appear to be those most distressed by the initial trauma; that there is no randomised controlled trial evidence to support the validity of group debriefing approaches; and that early intervention using Cognitive Behavioural techniques for those with clinically significant presentations appears the most promising approach. A generic set of guidelines for intervention following trauma is provided. PMID- 14523511 TI - [Pharmacological treatment of posttraumatic stress disorder]. AB - The authors present a review of pharmacotherapy for posttraumatic stress disorder (PTSD). Only a few controlled clinical trials have been carried out on PTSD, but there is a growing interest on this topic. Antidepressants, specially those with serotonergic activity, appear to provide effective pharmacotherapy for PTSD, as having either a primary therapeutic effect or in association with psychotherapy. PMID- 14523512 TI - [Posttraumatic stress disorder and major depression]. AB - Epidemiological studies clearly indicate that posttraumatic stress disorder (PTSD) is becoming a major health concern worldwide even if still poorly recognized and not well treated. PTSD commonly co-occurs with other psychiatric disorder, especially with major depression. However, the relationship between these disorders and the treatment of this complex clinical entity are only now being addressed in the specialized literature. Some authors argued that they are two distinct entities, whereas others defended the hypothesis that the high prevalence of this comorbidity may represent an artifact derived from the diagnostic criteria currently used. Regarding the treatment of PSTD comorbid with major depression, the available data from controlled studies are insufficient to point out for a specific approach, although some small trials reported the usefulness of combining selective serotonin reuptake inhibitors and cognitive therapy. PMID- 14523513 TI - [Posttraumatic stress disorder and bipolar mood disorder]. AB - Bipolar disorder (BD) is not only an endogenous condition. Severe negative life events have been shown to influence the development of the first episode and lifetime course of BD. Posttraumatic stress disorder (PTSD) is a severe and incapacitating mental condition that affect a significant proportion of the general population at some time in their lives. The concomitant presence of BD and PTSD has been shown to be more frequent than previously suggested and psychotic patients with trauma histories have a tendency to present more severe symptoms and are more proned to present substance use disorders. Trauma-related intrusive memories and nightmares of PTSD have been associated with mood changes. Also, kindling and behavioral sensitization have been proposed to explain the etiology and course of both disorders. Pharmacological approaches for this comorbidity are still based on empirical or not controlled approaches. In this article, we critically review the current literature regarding this co morbid condition, and highlight some aspects related to epidemiology, etiology, course and pharmacological treatment of both disorders. Overall, our review emphasizes the importance of systematically evaluate trauma histories in patients with BD. PMID- 14523514 TI - [Posttraumatic stress disorder: an evidence-based approach]. AB - The interventions used in the clinical management of posttraumatic stress disorder (PTSD) focus on: 1. Prevention of the development of the disorder, after a traumatic event 2. Treatment of the disorder, once it is already established, 3. Maintenance of long term functioning and quality of life. A variety of psychotherapies and pharmacological treatments have been proposed as therapeutic options in the treatment of PTSD. However, many of these treatment modalities lack scientific background. In this article authors present the treatment modalities of PTSD which are supported by scientific evidence and discuss its applications and drawbacks. PMID- 14523515 TI - Conversion of an amplified fragment length polymorphism marker into a co-dominant marker in the mapping of the Rph15 gene conferring resistance to barley leaf rust, Puccinia hordei Otth. AB - Leaf rust, caused by Puccinia hordei, is an important disease afflicting barley ( Hordeum vulgare) in many production regions of the world. The leaf rust resistance gene Rph15 was identified in an accession of wild barley ( Hordeum vulgare subsp. spontaneum) and is one of the most broadly effective resistance genes known. Using amplified fragment length polymorphism (AFLP) and simple sequence repeat markers, Rph15 was mapped to chromosome 2HS in an F(2) population derived from a cross between Bowman ( Rph15), a Bowman backcross-derived line carrying Rph15, and the susceptible cultivar Bowman. AFLP marker P13M40 co segregated with Rph15 in this mapping population and two others involving Bowman ( Rph15) and cultivars Proctor and Nudinka. The dominant AFLP marker P13M40 was converted to a co-dominant PCR-based marker that may be useful in breeding programs employing marker-assisted selection. The allelic relationship between Rph15 and the gene Rph16, also mapping to chromosome 2HS, was studied. The lack of segregation in F(2) progeny derived from the two resistance sources indicates that Rph15 and Rph16 are alleles of the same locus. PMID- 14523516 TI - Yr32 for resistance to stripe (yellow) rust present in the wheat cultivar Carstens V. AB - Stripe or yellow rust of wheat, caused by Puccinia striiformis f. sp. tritici, is an important disease in many wheat-growing regions of the world. A number of major genes providing resistance to stripe rust have been used in breeding, including one gene that is present in the differential tester Carstens V. The objective of this study was to locate and map a stripe rust resistance gene transferred from Carstens V to Avocet S and to use molecular tools to locate a number of genes segregating in the cross Savannah/Senat. One of the genes present in Senat was predicted to be a gene that is present in Carstens V. For this latter purpose, stripe rust response data from both seedling and field tests on a doubled haploid population consisting of 77 lines were compared to an available molecular map for the same lines using a non-parametric quantitative trait loci (QTL) analysis. Results obtained in Denmark suggested that a strong component of resistance with the specificity of Carstens V was located in chromosome arm 2AL, and this was consistent with chromosome location work undertaken in Australia. Since this gene segregated independently of Yr1, the only other stripe rust resistance gene known to be located in this chromosome arm, it was designated Yr32. Further QTLs originating from Senat were located in chromosomes 1BL, 4D, and 7DS and from Savannah on 5B, but it was not possible to characterize them as unique resistance genes in any definitive way. Yr32 was detected in several wheats, including the North American differential tester Tres. PMID- 14523517 TI - Candidate gene analysis of anthocyanin pigmentation loci in the Solanaceae. AB - Crop species in the Solanaceae, which includes tomato ( Lycopersicon esculentum), potato ( Solanum tuberosum), pepper ( Capsicum spp.), and eggplant ( S. melongena), exhibit natural variation in the types, levels, and tissue-specific expression patterns of anthocyanin pigments. While the identities of the genes underpinning natural variation in anthocyanin traits in these crops are largely unknown, many structural genes and regulators of anthocyanin biosynthesis have been isolated from the solanaceous ornamental species Petunia. To identify candidate genes that may correspond to loci controlling natural variation in the four crops, 13 anthocyanin-related genes were localized on a tomato F(2) genetic map. Gene map positions were then compared to mapped mutants in tomato and through comparative genetic maps to natural variants in potato, eggplant, and pepper. Similar map positions suggest that the tomato mutants anthocyaninless, entirely anthocyaninless, and anthocyanin gainer correspond to flavonoid 3'5' hydroxylase ( f3'5'h), anthocyanidin synthase, and the Petunia Myb domain trancriptional regulatory gene an2, respectively. Similarly potato R, required for the production of red pelargonidin-based pigments, P, required for production of purple delphinidin-based pigments, and I, required for tissue-specific expression in tuber skin, appear to correspond to dihydroflavonol 4-reductase, f3'5'h and an2, respectively. The map location of an2 also overlaps pepper A and eggplant fap10.1, lla10.1, lra10.1, sa10.1, pa10.1 and ca10.1, suggesting that a homologous regulatory locus has been subjected to parallel selection in the domestication of many solanaceous crops. To test the hypothesis that tomato anthocyaninless corresponds to f3'5'h, a portion of the gene was sequenced. A premature stop codon was observed in an anthocyaninless mutant, but not in wild type. PMID- 14523518 TI - Isolation and characterization of rice mutants compromised in Xa21-mediated resistance to X. oryzae pv. oryzae. AB - The rice gene, Xa21, confers resistance to diverse races of Xanthomonas oryzae pv. oryzae (Xoo) and encodes a receptor-like kinase with leucine-rich repeats in the extra-cellular domain. To identify genes essential for the function of the Xa21 gene, 4,500 IRBB21 ( Xa21 isogenic line in IR24 background) mutants, induced by diepoxybutane and fast neutrons, were screened against Philippine race six (PR6) Xoo for a change from resistance to susceptibility. From two greenhouse screens, 23 mutants were identified that had changed from resistant to fully (6) or partially (17) susceptible to PR6. All fully susceptible mutants carried rearrangements at the Xa21 locus as detected by PCR and Southern hybridization. For the partially susceptible mutants, no changes were detected at the Xa21 locus based on Southern and PCR analyses. However, two of these mutants were confirmed via genetic analysis to have mutations at the Xa21 locus. Partially susceptible mutants exhibited variation in level of susceptibility to different Xoo strains, suggesting that they may carry different mutations required for the Xa21-mediated resistance. The mutants identified in this study provide useful materials for dissecting the Xa21-mediated resistance pathway in rice. PMID- 14523519 TI - Mining data from potato pedigrees: tracking the origin of susceptibility and resistance to Verticillium dahliae in North American cultivars through molecular marker analysis. AB - Potato ( Solanum tuberosum L.) cultivated in North America is an autotetraploid species with a narrow genetic base. Most of the popular commercial cultivars are susceptible to Verticillium dahliae, a fungal pathogen causing Verticillium wilt disease, though some cultivars with relatively high resistance also exist. We have used the available pedigree information to track the origin of susceptibility and resistance to Verticillium wilt present in cultivated potatoes. One hundred thirty-nine potato cultivars and breeding selections were analyzed for resistance to the pathogen and for the presence of the microsatellite marker allele STM1051-193 that is closely linked to the resistance quantitative trait locus located on the short arm of chromosome 9. We detected an unusually high frequency of susceptible genotypes in the progeny descending from the breeding selection USDA X96-56. Molecular analysis revealed that USDA X96-56 does not have the STM1051-193 allele. Most of the first-generation progeny of this breeding selection also lack the allele. On the other hand, pedigree analysis indicated that breeding selection USDA 41956 often transfers V. dahliae resistance to its progeny. Molecular analysis detected presence of (at least) three STM1051-193 alleles in this breeding selection. These two genotypes (USDA X96-56 and USDA 41956) appear to have contributed greatly to the susceptibility or resistance, respectively, found in present commercial cultivars. Our results also indicate that the maturity class substantially affects the plant resistance response. In the intermediate to very late maturing class, the presence of the STM1051-193 allele significantly increases the resistance. Early to very early potatoes are usually more susceptible to the disease regardless of the allelic status, though the pattern of the allele effect is always the same. The results indicate that the STM1051-193 allele can be used for marker-assisted selection, but the potato maturity class also needs to be considered when making the final decision about the plant resistance level. PMID- 14523520 TI - Dissection of quantitative and durable leaf rust resistance in Swiss winter wheat reveals a major resistance QTL in the Lr34 chromosomal region. AB - The Swiss winter bread wheat cv. 'Forno' has a highly effective, durable and quantitative leaf rust ( Puccinia triticina Eriks.) resistance which is associated with leaf tip necrosis (LTN). We studied 240 single seed descent lines of an 'ArinaxForno' F(5:7 )population to identify and map quantitative trait loci (QTLs) for leaf rust resistance and LTN. Percentage of infected leaf area (%) and the response to infection (RI) were evaluated in seven field trials and were transformed to the area under the disease progress curves (AUDPC). Using composite interval mapping and LOD >4.4, we identified eight chromosomal regions specifically associated with resistance. The largest and most consistent leaf rust resistance locus was identified on the short arm of chromosome 7D (32.6% of variance explained for AUDPC_% and 42.6% for AUDPC_RI) together with the major QTL for LTN ( R(2)=55.6%) in the same chromosomal region as Lr34 ( Xgwm295). A second major leaf rust resistance QTL ( R(2)=28% and 31.5%, respectively) was located on chromosome arm 1BS close to Xgwm604 and was not associated with LTN. Additional minor QTLs for LTN (2DL, 3DL, 4BS and 5AL) and leaf rust resistance were identified. These latter QTLs might correspond to the leaf rust resistance genes Lr2 or Lr22 (2DS) and Lr14a (7BL). PMID- 14523521 TI - QTL and candidate genes phytoene synthase and zeta-carotene desaturase associated with the accumulation of carotenoids in maize. AB - Carotenoids are a class of fat-soluble antioxidant vitamin compounds present in maize ( Zea mays L.) that may provide health benefits to animals or humans. Four carotenoid compounds are predominant in maize grain: beta-carotene, beta cryptoxanthin, zeaxanthin, and lutein. Although beta-carotene has the highest pro vitamin A activity, it is present in a relatively low concentration in maize kernels. We set out to identify quantitative trait loci (QTL) affecting carotenoid accumulation in maize kernels. Two sets of segregating families were evaluated-a set of F2:3 lines derived from a cross of W64a x A632, and their testcross progeny with AE335. Molecular markers were evaluated on the F2:3 lines and a genetic linkage map created. High-performance liquid chromatography was performed to measure beta-carotene, beta-cryptoxanthin, zeaxanthin, and lutein on both sets of materials. Composite interval mapping identified chromosome regions with QTL for one or more individual carotenoids in the per se and testcross progenies. Notably QTL in the per se population map to regions with candidate genes, yellow 1 and viviparous 9, which may be responsible for quantitative variation in carotenoids. The yellow 1 gene maps to chromosome six and is associated with phytoene synthase, the enzyme catalyzing the first dedicated step in the carotenoid biosynthetic pathway. The viviparous 9 gene maps to chromosome seven and is associated with zeta-carotene desaturase, an enzyme catalyzing an early step in the carotenoid biosynthetic pathway. If the QTL identified in this study are confirmed, particularly those associated with candidates genes, they could be used in an efficient marker-assisted selection program to facilitate increasing levels of carotenoids in maize grain. PMID- 14523522 TI - Linkage disequilibrium mapping of a Verticillium dahliae resistance quantitative trait locus in tetraploid potato ( Solanum tuberosum) through a candidate gene approach. AB - We have used the linkage disequilibrium mapping method to test for an association between a candidate gene marker and resistance to Verticillium dahliae in tetraploid potato. A probe derived from the tomato Verticillium resistance gene ( Ve1) identified homologous sequences ( StVe1) in potato, which in a diploid population map to chromosome 9, in a position analogous to that of the tomato resistance gene. When a molecular marker closely linked (1.5 cM) to the homologues was used as a candidate gene marker on 137 tetraploid potato genotypes (mostly North American cultivars), the association between the marker and resistance was confirmed ( P<0.001). The amount of phenotypic variation in resistance explained by the allele of the STM1051 marker was greater than 10% and 25% in two subpopulations that were inferred from coancestry data matrix. Cloning of homologues from the highly resistant potato cv. Reddale indicates that the resistance quantitative trait locus (QTL) comprises at least an eleven-member family, encoding plant-specific leucine-rich repeat proteins highly similar to the tomato Ve genes. The sequence analysis shows that all homologues are uninterrupted open reading frames and thus represent putative functional resistance genes. This is the first time that the linkage disequilibrium method has been used to find an association between a resistance gene and a candidate gene marker in tetraploid potato. We have shown that it is possible to map QTL directly on already available potato cultivars, without developing a new mapping population. PMID- 14523523 TI - Accumulation, assembly, and digestibility of amarantin expressed in transgenic tropical maize. AB - An amaranth ( Amaranthus hypochondriacus) 11S globulin cDNA, encoding one of the most important storage proteins (amarantin) of the seed, with a high content of essential amino acids, was used in the transformation of CIMMYT tropical maize genotype. Constructs contained the amarantin cDNA under the control of a tissue specific promoter from rice glutelin-1 ( osGT1) or a constitutive ( CaMV 35S) promoter with and without the first maize alcohol dehydrogenase intron ( AdH). Southern-blot analysis confirmed the integration of the amarantin cDNA, and copy number ranged from one to more than ten copies per maize genome. Western-blot and ultracentrifugation analyses of transgenic maize indicate that the expressed recombinant amarantin precursors were processed into the mature form, and accumulated stably in maize endosperm. Total protein and some essential amino acids of the best expressing maize augmented 32% and 8-44%, respectively, compared to non-transformed samples. The soluble expressed proteins were susceptible to digestion by simulated gastric and intestinal fluids, and it is suggested that they show no allergenic activity. These findings demonstrate the feasibility of using genetic engineering to improve the amino acid composition of grain crops. PMID- 14523524 TI - Gene flow among different taxonomic units: evidence from nuclear and cytoplasmic markers in Cedrus plantation forests. AB - Hybridization and introgression are important natural evolutionary processes that can be successfully investigated using molecular markers and open- and controlled pollinated progeny. In this study, we collected open-pollinated seeds from Cedrus atlantica, Cedrus libani and C. libani x C. atlantica hybrids from three French plantation forests. We also used pollen from C. libani and Cedrus brevifolia to pollinate C. atlantica trees. The progeny were analyzed using three different types of molecular markers: RAPDs, AFLPs and cpSSRs. Chloroplast DNA was found to be paternally inherited in Cedrus from the progeny of controlled-crosses. Heteroplasmy, although possible, could not be undoubtedly detected. There was no indication of strong reproductive isolating barriers among the three Mediterranean Cedrus taxa. Gene flow between C. atlantica and C. libani accounted for 67 to 81% of viable open-pollinated seedlings in two plantation forests. We propose that Mediterranean Cedrus taxa should be considered as units of a single collective species comprising two regional groups, North Africa and the Middle East. We recommend the use of cpSSRs for monitoring gene flow between taxa in plantation forests, especially in areas where garden specimens of one species are planted in the vicinity of selected seed-stands and gene-conservation reserves of another species. PMID- 14523525 TI - Complex population genetic structure in the endemic Canary Island pine revealed using chloroplast microsatellite markers. AB - The Canary archipelago, located on the northwestern Atlantic coast of Africa, is comprised of seven islands aligned from east to west, plus seven minor islets. All the islands were formed by volcanic eruptions and their geological history is well documented providing a historical framework to study colonization events. The Canary Island pine ( Pinus canariensis C. Sm.), nowadays restricted to the westernmost Canary Islands (Gran Canaria, Tenerife, La Gomera, La Palma and El Hierro), is considered an old (Lower Cretaceous) relic from an ancient Mediterranean evolutionary centre. Twenty seven chloroplast haplotypes were found in Canary Island pine but only one of them was common to all populations. The distribution of haplotypic variation in P. canariensis suggested the colonization of western Canary Islands from a single continental source located close to the Mediterranean Basin. Present-day populations of Canary Island pine retain levels of genetic diversity equivalent to those found in Mediterranean continental pine species, Pinus pinaster and Pinus halepensis. A hierarchical analysis of variance (AMOVA) showed high differentiation among populations within islands (approximately 19%) but no differentiation among islands. Simple differentiation models such as isolation by distance or stepping-stone colonization from older to younger islands were rejected based on product-moment correlations between pairwise genetic distances and both geographic distances and population-age divergences. However, the distribution of cpSSR diversity within the islands of Tenerife and Gran Canaria pointed towards the importance of the role played by regional Pliocene and Quaternary volcanic activity and long-distance gene flow in shaping the population genetic structure of the Canary Island pine. Therefore, conservation strategies at the population level are strongly recommended for this species. PMID- 14523526 TI - Phenotypic, genetic and molecular characterization of a maize low phytic acid mutant (lpa241). AB - Phytic acid, myo-inositol 1,2,3,4,5,6-hexakisphosphate, is the major storage compound of phosphorous (P) in plants, predominantly accumulating in seeds (up to 4-5% of dry weight) and pollen. In cereals, phytic acid is deposited in embryo and aleurone grain tissues as a mixed "phytate" salt of potassium and magnesium, although phytates contain other mineral cations such as iron and zinc. During germination, phytates are broken down by the action of phytases, releasing their P, minerals and myo-inositol which become available to the growing seedling. Phytic acid represents an anti-nutritional factor for animals, and isolation of maize low phytic acid ( lpa) mutants provides a novel approach to study its biochemical pathway and to tackle the nutritional problems associated with it. Following chemical mutagenesis of pollen, we have isolated a viable recessive mutant named lpa 241 showing about 90% reduction of phytic acid and about a tenfold increase in seed-free phosphate content. Although germination rate was decreased by about 30% compared to wild-type, developement of mutant plants was apparentely unaffected. The results of the genetic, biochemical and molecular characterization experiments carried out by SSR mapping, MDD-HPLC and RT-PCR are consistent with a mutation affecting the MIPS1S gene, coding for the first enzyme of the phytic acid biosynthetic pathway. PMID- 14523527 TI - Grass consensus STS markers: an efficient approach for detecting polymorphism in Lolium. AB - For ryegrass and many forage crops, characterization of varieties is often difficult for two reasons: few of discriminant morphological traits and a great within-varieties variation. Futhermore, few molecular markers are publicly accessible. In this paper we describe two approaches for the development of 42 sequence-tagged-site (STS) markers. Firstly, 14 STS markers were developed from Lolium sequences found in data bases. Secondly, 28 STS markers were developed from sequences found in related species of Gramineae. Out of 42 STS markers developed, 85.8% yielded successfull amplification and 62% revealed a high level of polymorphism with an average of five alleles per locus. The analysis of amplicons reveals a high STS marker specificity, a high conservation in gene structure and a strong intron sequence homology between allelic forms. Moreover, the majority of the STS markers can be considered as "universal markers" because 81% of these STS markers amplified successfully across 20 related grass species. These results permit us to consider the use of these markers in synteny studies. PMID- 14523528 TI - Molecular cytogenetic analysis of genome structure in Lupinus angustifolius and Lupinus cosentinii. AB - Molecular cytogenetic analysis of Lupinus angustifolius and Lupinus cosentinii was performed using flow cytometry, fluorescence in situ hybridisation (FISH) and differential chromosome staining. Genome size was determined as 2.07 pg for L. angustifolius and 1.54 pg for L. cosentinii. Analysis of nuclear DNA amount in cells during plant development has shown endopolyploidisation in different organs. The highest level of endopolyploidy was in cotyledons and reached 32C in L. angustifolius and 64C in L. cosentinii. Both of the investigated Lupinus species belong to the polysomatic type of plants. Double FISH with rDNA probes provided chromosomal landmarks for 10 out of 40 chromosomes for L. angustifolius and 8 out of 32 chromosomes for L. cosentinii. In L. angustifolius, the number and localisation of 25S rDNA hybridisation signals precisely corresponded to the chromomycin A3 (CMA(+)) bands, while in L. cosentinii both 25S and 5S rDNA loci overlapped with CMA(+) bands. Silver staining revealed that only 45S rRNA genes located in secondary constriction regions were transcriptionally active. FISH with Arabidopsis-type telomeric arrays revealed the presence of signals at termini of all chromosomes. Despite the application of different DNA probes for FISH and different chromosome staining, a relatively small proportion of chromosomes in the Lupinus karyotypes can be distinguished. Identification of all chromosomes requires the use of more chromosome-specific markers. PMID- 14523529 TI - Allele-specific PCR detection of sweet cherry self-incompatibility (S) alleles S1 to S16 using consensus and allele-specific primers. AB - PCR-based identification of all 13 known self-incompatibility (S) alleles of sweet cherry is reported. Two pairs of consensus primers were designed from our previously published cDNA sequences of S(1) to S(6) S-RNases, the stylar components of self-incompatibility, to reveal length variation of the first and the second introns. With the exception of the first intron of S(13), these also amplified S(7) to S(14) and an allele previously referred to as S(x), which we now label S(16). The genomic PCR products were cloned and sequenced. The partial sequence of S(11) matched that of S(7) and the alleles were shown to have the same functional specificity. Allele-specific primers were designed for S(7) to S(16), so that allele-specific primers are now available for all 13 S alleles of cherry (S(8), S(11) and S(15) are duplicates). These can be used to distinguish between S alleles with introns of similar size and to confirm genotypes determined with consensus primers. The reliability of the PCR with allele specific primers was improved by the inclusion of an internal control. The use of the consensus and allele-specific primers was demonstrated by resolving conflicting genotypes that have been published recently and by determining genotypes of 18 new cherry cultivars. Two new groups are proposed, Group XXIII (S(3) S(16)), comprising 'Rodmersham Seedling' and 'Strawberry Heart', and Group XXIV (S(6) S(12)), comprising 'Aida' and 'Flamentiner'. Four new self compatibility genotypes, S(3) S(3)', S(4)' S(6), S(4)' S(9) and S(4)' S(13), were found. The potential use of the consensus primers to reveal incompatibility alleles in other cherry species is also demonstrated. PMID- 14523530 TI - [Prognostic significance of immunohistochemical studies]. PMID- 14523531 TI - [Tinnitus and craniomandibular dysfunction]. PMID- 14523532 TI - [Tolerance and effectiveness of oxymetazoline and xylometazoline in treatment of acute rhinitis]. AB - BACKGROUND: Local alpha-sympathomimetics in hydrous solution are well known in the therapy of acute rhinitis and sinusitis. However, added preservatives like benzalkonium chloride have a negative effect on compatibility. PATIENTS AND METHOD: A total of 307 patients with acute rhinitis entered the study. The treatment with oxymetazoline with preservative, oxymetazoline without preservative and xylometazoline with preservative was evaluated. RESULTS: This randomised, double-blind, multi-centered, verum-controlled tolerance study confirmed that the local sympathomimetics oxymetazoline and xylometazoline are well tolerated in the treatment of acute rhinitis. When evaluated according to the parameters "feeling of dryness in nasal mucosa" and "burning sensation", the Nasivin sanft 0.05% spray, which contains the active agent oxymetazoline without preservatives, proved to be considerably superior to preparations containing the preservative benzalkonium chloride. CONCLUSION: Preparations without preservatives should be the preferred choice of treatment for acute rhinitis. PMID- 14523533 TI - [Immunohistochemical determination of cell cycle regulatory proteins. No prognostic significance in advanced squamous epithelial carcinomas of the head neck area]. AB - BACKGROUND: In recent years, a number of molecular markers have been tested for their potential to predict the outcome after radiotherapy or radiochemotherapy in patients with advanced head and neck squamous cell carcinomas (HNSCC). These studies have produced controversial results. PATIENTS AND METHODS: The expression patterns of the proteins p53, pRb, cyclinD1, cdk4, p21(CIP1/WAF1), p16(INK4a), bcl-2, and mib-1/ki-67 were analyzed in pretreatment tumor biopsies of 53 patients with advanced nonresectable head and neck squamous cell carcinomas (mainly UICC IV). The patterns obtained were compared with clinical outcome after accelerated radiochemotherapy with carboplatin and 5-fluorouracil or accelerated fractionated radiotherapy alone, respectively. RESULTS: The expression patterns of the proteins examined did not correlate with response to therapy or with further clinical course. CONCLUSION: In patients with advanced head and neck squamous cell carcinomas, the prognostic relevance of pretreatment expression patterns of the proteins investigated in this study, particularly p53, cyclinD1, and cdk4, has to be questioned. PMID- 14523534 TI - [Reliability and general practice value of 2 rapid Streptococcus A tests]. AB - Rapid streptococcal-A-antigen detection assays have good specificity (over 90 percent) but moderate sensitivity (between 80 and 90 percent), when the tests are compared with a standard throat culture. Contradictory results have been found for one of the more recent tests, the optical immune assay Strep A OIA MAX, while for 6 years, we have been using the immune assay Strep A Plus. Results of the optical immunoassay and the conventional immune assay Strep A Plus were compared in 65 patients with acute pharyngitis. A standard culture was used as reference and confirmed by enhanced broth culturing and nucleic acid hybridization assay (Gen-Probe) when the two detection assays produced contradictory results. While both assays were equally sensitive (78.3%), Strep A OIA MAX and Strep A Plus had a similar specificity of 95.2% and 100%, respectively. Four and nine steps were required for Strep A Plus and for Strep A OIA MAX test procedures, respectively with results being available in 4-5 min and in 9-10 min, respectively. We conclude that both rapid immunoassays have a similar reliability while the handling of the Strep A Plus is much simpler than the handling of the Strep A OIA MAX. Neither rapid immunoassays are sensitive enough to eliminate the need for backup cultures. PMID- 14523536 TI - [Pigment spots on the tongue]. PMID- 14523535 TI - [Brain abscesses after extracranial infections of the head and neck area]. AB - The authors report on 20 immunocompetent patients with brain abscess after 12 cases of middle ear, seven tooth and a single frontal sinus infection. The clinical aspects, hematochemical and microbiological data, the role of imaging diagnostics (CT, MR) and the type of treatment are analysed. Neurosurgery was performed on 17 patients (85%), eight of whom subsequently underwent evacuation of the primary source of infection (four mastoidectomies, two timpanoplasties, two tooth extractions). Mastoidectomy was eventually carried out on one of the three patients who did not undergo neurosurgery. Microbiological diagnosis was possible in nine patients through culture examination: Proteus mirabilis in three cases, Peptostreptococcus sp. in two, Micrococcus varians, Proteus vulgaris, Streptococcus sanguis and Streptococcus viridans not typed in single cases. The pus was sterile in eight patients (47.1% of those operated). An association of two antimicrobial agents was used in 18 patients, while in two cases monotherapy was preferred, based on the isolated bacteria. Treatment lasted on average 38 days. The most frequently used therapy regimen (75%) was the association of a beta-lactam drug with chloramphenicol or metronidazole. Therapy was successful in 19/20 patients; one patient died. There was no significant difference in prognostic terms with regard to sex, age, duration of symptoms prior to diagnosis, clinical picture at onset, number and size of abscesses or type of treatment. Recognising the first clinical signs and symptoms (headache, fever, alterations in consciousness, focal neurological deficit, epileptic seizures) is extremely important for prompt diagnosis of brain abscess. PMID- 14523538 TI - [Impalement injury of the neck]. AB - In comparison to the United States or South Africa, penetrating injuries of the neck are rare in Europe. Most of these traumas are due to sharp perforation mechanisms. We report on a 43-year-old man who was admitted to the emergency room because of an impressive transcervical penetrating neck trauma inflicted by a chisel. He survived the trauma since the chisel missed all important structures of the neck. The diagnostic strategy to evaluate the dimension of the trauma was primarily based on endoscopic and surgical exploration. PMID- 14523537 TI - [Progressive paralysis caused by radiation-induced cervical malignant peripheral nerve sheath tumor]. AB - HISTORY AND DIAGNOSIS: A 59-year old engineer was admitted to the hospital because of pain in his right collar region and the onset of incomplete paresis of the right arm. Magnetic resonance tomography displayed an advanced tumour arising from the right paravertebral soft tissue. Histological examination revealed a malignant peripheral nerve sheath tumor (MPNST). Thirteen years before admission, the patient had a right-sided tumor-tonsillectomy of a squamous cell carcinoma and local radiation of a cystic squamous cell carcinoma in the ipsilateral cervical soft tissue. CLINICAL COURSE AND THERAPY: In the following course, progressive neurological symptoms occurred including beginning paraplegia, right phrenic paralysis and a severe concomitant pain syndrome. Due to the location and advanced tumor state, surgical treatment was not performed and palliative chemotherapy remained ineffective. Three months later the patient died due to rapidly progressive neurological failure. CONCLUSION: MPNST represents a rare entity which has been related to postoperative radiation. Unusual neurological symptoms in anatomical regions of former radiation should therefore include neurogenic secondary malignancies in the differential diagnosis for early surgical intervention. PMID- 14523539 TI - [Reinke edema--helpful in unilateral recurrent laryngeal nerve paralysis?]. AB - We report on a 57-year-old teacher who consulted us for her known unilateral recurrent nerve paralysis subsequent to strumectomy. Video laryngoscopic and stroboscopic examinations revealed Reinke's edema in addition to the left-sided immobile vocal fold with maintenance of good vocal function. Because of the left sided Reinke's edema, complete glottal closure was possible although the left vocal ligament remained slack. Surgical removal of the vocal fold edema in this case would presumably result in considerable impairment of vocal function. This example illustrates that in selected cases Reinke's edema can contribute to maintenance of phonation in unilateral vocal fold paralysis. In these cases, indication for edema surgery should be very restrictive and should always take the vocal function into consideration. PMID- 14523540 TI - [Active language development in children with severe hearing loss and deafness in relation to technical auditory management]. AB - The extent of vocabulary depends on the success of rehabilitation in hearing, language and speech. In hearing impaired and deaf children, it is important to select the optimum technical device-hearing aid or cochlear implant. This study was performed on 30 children with hearing aids or cochlear implants using the Test of Expressive Vocabulary for Children Aged 3-6 years (Kiese-Himmel and Kozielski). The development of vocabulary was recorded over 12 months. The results were analysed dependent on the degree of hearing loss, age at primary diagnosis, duration of treatment and kind of hearing aid used. These were compared with the reference data from a normally hearing population. Cochlear implant users had better language development than hearing aid users despite a lower grade of hearing loss in the hearing aid group. In general, the extent of the expressive vocabulary was less in hearing impaired children than in the reference population. However, in terms of hearing age, the language development of some cochlear implant users was similar to that of normal children. PMID- 14523541 TI - [Mumps]. PMID- 14523542 TI - Stochastic gene expression during cell differentiation: order from disorder? PMID- 14523543 TI - Neuroprotective effects of Ginkgo biloba extract. AB - Ginkgo biloba extract has been therapeutically used for several decades to increase peripheral and cerebral blood flow as well as for the treatment of dementia. The extract contains multiple compounds such as flavonoids and terpenoids that are thought to contribute to its neuroprotective and vasotropic effects. In this review, we summarize the experimental results on the mechanism of neuroprotection induced by standardized extract of Ginkgo biloba leaves (EGb 761) and its constituents. The effects described mostly in animals include those on cerebral blood flow, neurotransmitter systems, cellular redox state and nitric oxide level. Furthermore, we discuss the current status of clinical trials as well as undesired side effects of EGb 761. PMID- 14523545 TI - Cbl signaling networks in the regulation of cell function. AB - Cbl proteins control multiple cellular processes by acting as ubiquitin ligases and multifunctional adaptor molecules. They are involved in the control of cell proliferation, differentiation and cell morphology, as well as in pathologies such as autoimmune diseases, inflammation and cancer. Here we review recent advances in understanding the role of Cbl and the importance of a growing repertoire of Cbl-interacting proteins in the regulation of signaling pathways triggered by growth factors, antigens, cell adhesion, cytokines and hormones. We also address key issues of the nature of proteins that bind Cbl in particular cells, where they are located, and how they are altered or traffic within cells upon stimulation. It is becoming obvious that temporal and spatial changes in Cbl signaling networks are essential for the control of physiological processes in a variety of cells and organs and that their deregulation can result in the development of human diseases. PMID- 14523544 TI - Recognition of bacterial peptidoglycan by the innate immune system. AB - The innate immune system recognizes microorganisms through a series of pattern recognition receptors that are highly conserved in evolution. Peptidoglycan (PGN) is a unique and essential component of the cell wall of virtually all bacteria and is not present in eukaryotes, and thus is an excellent target for the innate immune system. Indeed, higher eukaryotes, including mammals, have several PGN recognition molecules, including CD14, Toll-like receptor 2, a family of peptidoglycan recognition proteins, Nod1 and Nod2, and PGN-lytic enzymes (lysozyme and amidases). These molecules induce host responses to microorganisms or have direct antimicrobial effects. PMID- 14523546 TI - The vault complex. AB - Vaults are large ribonucleoprotein particles found in eukaryotic cells. They are composed of multiple copies of a Mr 100,000 major vault protein and two minor vault proteins of Mr 193,000 and 240,000, as well as small untranslated RNAs of 86-141 bases. The vault components are arranged into a highly characteristic hollow barrel-like structure of 35 x 65 nm in size. Vaults are predominantly localized in the cytoplasm where they may associate with cytoskeletal elements. A small fraction of vaults are found to be associated with the nucleus. As of yet, the precise cellular function of the vault complex is unknown. However, their distinct morphology and intracellular distribution suggest a role in intracellular transport processes. Here we review the current knowledge on the vault complex, its structure, components and possible functions. PMID- 14523547 TI - Fatty acid metabolism in Saccharomyces cerevisiae. AB - Peroxisomes are essential subcellular organelles involved in a variety of metabolic processes. Their importance is underlined by the identification of a large group of inherited diseases in humans in which one or more of the peroxisomal functions are impaired. The yeast Saccharomyces cerevisiae has been used as a model organism to study the functions of peroxisomes. Efficient oxidation of fatty acids does not only require the participation of peroxisomal enzymes but also the active involvement of other gene products. One group of important gene products in this respect includes peroxisomal membrane proteins involved in metabolite transport. This overview discusses the various aspects of fatty acid beta-oxidation in S. cerevisiae. Addressed are the various enzymes and their particular functions as well as the various transport mechanisms to take up fatty acids into peroxisomes or to export the beta-oxidation products out of the peroxisome to mitochondria for full oxidation to CO2 and H2O. PMID- 14523548 TI - Protein folding revisited. A polypeptide chain at the folding-misfolding nonfolding cross-roads: which way to go? AB - The structure-function paradigm claims that a specific function of a protein is determined by its unique and rigid three-dimensional (3D) structure. Thus, following its biosynthesis on the ribosome, a protein must fold to be functional. This idea represents one of the cornerstones of modern biology. Numerous cases when, due to the effect of environmental factors or because of genetic defects (mutations), a polypeptide chain has lost its capability to gain a proper functional 3D structure (i.e. became misfolded), seem to confirm this concept. Consequences of such misfolding are well known and represent lost of function, aggregation, development of conformational disorders and cell death. However, the recent revelation of countless examples of intrinsically disordered proteins has cast doubt on the general validity of the structure-function paradigm and revealed an intriguing route of functional disorder. Thus, in a living cell, a polypeptide chain chooses between three potential fates - functional folding, potentially deadly misfolding and mysterious nonfolding. This choice is dictated by the peculiarities of amino acid sequence and/or by the pressure of environmental factors. The aim of the present review is to outline some interesting features of these three routes. PMID- 14523549 TI - Structure and function of desmosomal proteins and their role in development and disease. AB - Desmosomes represent major intercellular adhesive junctions at basolateral membranes of epithelial cells and in other tissues. They mediate direct cell-cell contacts and provide anchorage sites for intermediate filaments important for the maintenance of tissue architecture. There is increasing evidence now that desmosomes in addition to a simple structural function have new roles in tissue morphogenesis and differentiation. Transmembrane glycoproteins of the cadherin superfamily of Ca(2+)-dependent cell-cell adhesion molecules which mediate direct intercellular interactions in desmosomes appear to be of central importance in this respect. The complex network of proteins forming the desmosomal plaque associated with the cytoplasmic domain of the desmosomal cadherins, however, is also involved in junction assembly and regulation of adhesive strength. This review summarizes the structural features of these desmosomal proteins, their function during desmosome assembly and maintenance, and their role in development and disease. PMID- 14523550 TI - Neuroreplacement therapy and stem cell biology under disease conditions. AB - Recent advances in stem cell technology are expanding our ability to replace a variety of cells throughout the body. In the past, neurological diseases caused by the degeneration of neuronal cells were considered incurable because of a long held 'truism'; neurons do not regenerate during adulthood. However, this statement has been challenged, and we have now found much evidence that the brain is indeed capable of regenerating neurons after maturing. Based on this new concept, researchers have shown neural differentiation of stem cells and recovery of function following transplantation of these cells into the brain. These results may promise a bright future for clinical applications of stem cell strategies in neurological diseases; however, we must consider the pathophysiological environments of individual diseases that may affect stem cell biology. Before we begin to develop clinical applications, we must consider environmental factors that have not been discussed in the current preclinical studies. Here, we study cases of Alzheimer's disease and schizophrenia and discuss the effects of environmental factors under disease conditions. PMID- 14523552 TI - Can different genetic changes characterize histogenetic subtypes and biologic behavior in sporadic malignant melanoma of the skin? AB - In sporadic malignant melanoma, different chromosomal regions with nonrandom aberrations have been discovered, including 1p36, 6q, 9p and 10q. First results provide a genetic basis for the concept of primarily vertical, biologically aggressive melanomas and radial growing, mostly benign melanomas. These are mainly represented by nodular melanoma (NM) and early superficial spreading melanoma (SSM), respectively. Deletions in 1p36 could be found only in NMs and metastatic melanoma. Aberrations of chromosome 10 occur predominantly in NMs, whereas deletions on chromosome 9 are more frequent in SSMs. Despite a variety of genes tested, neither a tumor suppressor gene with importance in all malignant melanomas of the skin nor one clearly defining the transition from the radial growth phase to the vertical growth phase has been determined. Nevertheless, the pattern of genetic alterations may soon lead to finding such genes and development of drugs targeting these genes or their products, which would be of great benefit to melanoma patients. PMID- 14523551 TI - Immunology and functional genomics of Behcet's disease. AB - Behcet's disease (BD) is a multisystemic inflammatory disorder. Although the cause and pathogenesis of BD are still unclear, there is evidence for genetic, immunologic and infectious factors at the onset or in the course of BD. This review focuses on the functional genomics and immunology of BD. HLA-B51 is the major disease susceptibility gene locus in BD. An increased number of gammadelta T cells in the peripheral blood and in the involved tissues have been reported. However, the T cells at the sites of inflammation appear to be a phenotypically distinct subset. There is also a significant gammadelta T cell proliferative response to mycobacterial 65-kDa heat shock protein peptides. Homologous peptides derived from the human 60-kDa heat shock protein were observed in BD patients. There is evidence that natural killer T cells may also play a role in BD. PMID- 14523553 TI - Soluble proteins of chemical communication in the social wasp Polistes dominulus. AB - Members of the odorant-binding protein (OBP) and chemosensory protein (CSP) families were identified and characterised in the sensory tissues of the social wasp Polistes dominulus (Hymenoptera: Vespidae). Unlike most insects so far investigated, OBPs were detected in antennae, legs and wings, while CSPs appeared to be preferentially expressed in the antennae. The OBP is very different from the homologous proteins of other Hymenopteran species, with around 20% of identical residues, while the CSP appears to be much better conserved. Both OBP and CSP, not showing other post-translational modifications apart from disulphide bridges, were expressed with high yields in a bacterial system. Cysteine pairing in the recombinant and native proteins follows the classical arrangements described for other members of these classes of proteins. OBPs isolated from the wings were found to be associated with a number of long-chain aliphatic amides and other small organic molecules. Binding of these ligands and other related compounds was measured for both recombinant OBP and CSP. PMID- 14523554 TI - A small chimerically bifunctional monomeric protein: Tapes japonica lysozyme. AB - The lysozyme of the marine bilave Tapes japonica (13.8 kDa) is a novel protein. The protein has 46% homology with the destabilase from medicinal leech that has isopeptidase activity. Based on these data, we confirmed hydrolysis activity of T. japonica lysozyme against three substrates: L-gamma-Glu-pNA, D-gamma-Glu-pNA, and epsilon-(gamma-Glu)-L-Lys. The optimal pH of chitinase and isopeptidase activity was 5.0 and 7.0, respectively. The isopeptidase activity was inhibited with serine protease inhibitor, but the lytic and chitinase activities were not. Moreover, only isopeptidase activity is decreased by lyophilization, but lytic and chitinase activities were not. We conclude that T. japonica lysozyme expresses isopeptidase and chitinase activity at different active sites. PMID- 14523555 TI - Notable diversity in hemoglobin expression patterns among species of the deep-sea clam, Calyptogena. AB - The deep-sea clams Calyptogena nautilei and C. tsubasa, which live in the cold seep area at a depth of 3570 m in the Nankai Trough, Japan, have abundant hemoglobins (Hbs) in erythrocytes, similar to other Calyptogena species. We determined the cDNA-derived amino acid sequences of Hbs from two Calyptogena species. C. tsubasa was found to contain two dimeric Hbs, Hb I consisting of 145 amino acid residues and Hb II with 137 residues, similar to known Hbs from C. soyoae and C. kaikoi. Sequence identity was over 90% among the orthologous chains of Calyptogena Hbs. On the other hand, surprisingly, C. nautilei contained two monomeric Hbs, Hb III containing 141 residues and Hb IV with 134 residues. In addition, Hbs III and IV showed only 33-42% sequence identity with Hbs I and II from other Calyptogena species. The distal (E7) histidine, one of the functionally important residues of the heme protein, is replaced by glutamine in all Hb chains of Calyptogena species. A phylogenetic analysis indicated that C. nautilei Hb III is closer to Hb I from other Calyptogena species. We suppose that a Hb gene was duplicated at least three times in an immediate ancestor of Calyptogena and, presumably depending on physiological conditions different Hb sets are being expressed: dimeric Hbs I and II in C. soyoae, C. kaikoi and C. tsubasa, and monomeric Hbs III and IV in C. nautilei. PMID- 14523556 TI - Potent thermogenic action of triiodothyroacetic acid in brown adipocytes. AB - Triiodothyroacetic acid (TRIAC) is a triiodothyronine (T3) metabolite with high affinity for T3 nuclear receptors. We compared the thermogenic action of TRIAC versus T3 in brown adipocytes, by studying target genes known to mediate thermogenic action: uncoupling protein 1 (UCP-1), a marker of brown adipocytes, and type II-5'deiodinase (D2), which provides the T3 required for thermogenesis. TRIAC is 10-50 times more potent than T3 at increasing the adrenergic induction of UCP-1 mRNA and D2 activities. TRIAC action on UCP-1 is exerted at the transcriptional level. In the presence of an adrenergic stimulus, TRIAC is also more potent than T3, inducing lipoprotein lipase mRNA and 5 deiodinase (D3) activity and mRNA. Maximal effects occur at very low concentrations (0.2 nM). The greater potency of TRIAC is not due to preferential cellular or nuclear uptake. Therefore, TRIAC is a potent thermogenic agent that might increase energy expenditure and regulate T3 production in brown adipocytes. PMID- 14523557 TI - Nitric oxide mediates histamine induced down-regulation of H2 receptor mRNA and internalization of the receptor protein (R1). AB - During agonist-dependent long-term stimulation of cells, histamine receptor subtypes are frequently down-regulated. However, the mechanisms underlying the modulation of receptor expression during long-term histamine stimulation have yet to be resolved. Based on our recently reported results showing an H1-mediated down-regulation of histamine H2 receptor mRNA in endothelial cells, our aim was to characterize the mechanism controlling rapid and long-term histamine-mediated modulation of H2 receptor expression in more detail. We were able to show that the histamine-induced down-regulation of H2 receptor mRNA and cell surface expression lasting for 24 h was accompanied by augmentation of the receptor protein level in the cytoplasmatic fraction of endothelial cells for this time period. Furthermore, changes in receptor protein levels in whole-cell lysate were negligible, indicating that the rapid and prolonged modulation of cell surface H2 receptor levels by histamine was regulated solely via internalization. The role of nitric oxide (NO) as a key mediator in histamine-stimulated cell responses was underlined by subsequent studies showing the attenuation of histamine-induced H2 receptor mRNA down-regulation and protein trafficking following NO synthase isozyme inhibition. PMID- 14523558 TI - Gender- and site-related effects on lipolytic capacity of rat white adipose tissue. AB - Gender- and site-related differences in the lipolytic capacity, at the different steps of the adrenergic pathway, in gonadal and inguinal white adipose tissue (WAT), were assessed by studying alpha2A-adrenergic receptor (AR), beta3-AR and hormone-sensitive lipase (HSL) protein levels, and by determining the lipolytic response to different agents. Gonadal WAT showed a lower alpha2A/beta3-AR ratio, a greater lipolytic capacity in response to AR agonists, and higher HSL activity and protein levels than inguinal WAT. In female rats, we found greater alpha2A-AR protein levels and alpha2A/beta3-AR ratio compared to their male counterparts, but, on the other hand, a higher lipolytic response to beta-AR agonists and a greater lipolytic capacity at the postreceptor level, including a more activated HSL protein. Thus, the lipolytic capacity was clearly higher in gonadal than in inguinal WAT, at the different steps of the adrenergic pathway studied. Moreover, in both tissues, females showed a greater inhibition of lipolysis via alpha2-AR, which was counteracted by the higher lipolytic capacity at the postreceptor level. PMID- 14523559 TI - Differential regulation of the Sir2 histone deacetylase gene family by inhibitors of class I and II histone deacetylases. AB - The Sir2 histone deacetylase gene family consists of seven mammalian sirtuins (SIRTs) which are NAD-dependent histone/protein deacetylases. Sir2 proteins regulate, for instance, genome stability by chromatin silencing in yeast. In mammals, their function is still largely unknown. Due to the NAD+ dependency, Sir2 might be the link between metabolic activity and histone/protein acetylation. Regulation of gene expression also seems to play an important role in Sir2 functions, since increasing the dosage of Sir2 genes increases genome stability in yeast and Caenorhabditis elegans. We observed that the modification of histone/protein acetylation status by several class I and II histone deacetylase (HDAC) inhibitors induces differential changes in gene expression profiles of seven SIRT mRNAs in cultured neuronal cells. SIRT2, SIRT4 and SIRT7 were upregulated, whereas SIRT1, SIRT5 and SIRT6 were downregulated by trichostatin A (TSA) and n-butyrate. The upregulation of SIRT mRNAs was inhibited by actinomycin D. Interestingly, the regulation of SIRT mRNAs was highly similar both in mouse Neuro-2a neuroblastoma cells and post-mitotic rat primary hippocampal and cerebellar granule neurons. Using a chromatin immunoprecipitation technique, we showed that the upregulation of SIRT2 expression with TSA is related to the hyperacetylation of DNA-bound histone H4 within the first 500 bp upstream of the transcription start site of the SIRT2 gene. Chemically different types of HDAC inhibitors, such as TSA, apicidin, SAHA, M344 and n-butyrate induced remarkably similar responses in SIRT1-7 mRNA expression patterns. Differential responses in SIRT mRNA expression profiles indicate that the expression of the Sir2 family of genes is selectively regulated and dependent on histone/protein acetylation status. PMID- 14523560 TI - Identification of the region in Escherichia coli DnaA protein required for specific recognition of the DnaA box. AB - DnaA protein binds specifically to a 9-base- pair motif called the DnaA box. Domain IV comprises 94 amino acid residues and is required for DNA binding. Using nuclear magnetic resonance analysis, we investigated the interaction between DnaA domain IV and both a DnaA box and a non-specific oligonucleotide that has a reduced affinity for DnaA. The 1H-15N HSQC spectrum of DnaA domain IV showed prominent chemical shift perturbations on six residues (Arg399, Ala404, Leu422, Asp433, Thr435 and Thr436) in the presence of the DnaA box. Through homology modeling, we located all of these residues on one side surface of the DnaA domain IV molecule. Moreover, we compared the chemical shift perturbation of the 1H-15N HSQC spectrum in the presence of the DnaA box with that in the presence of a non specific oligonucleotide, and the results suggested that Leu422 imparts specificity in binding with the DnaA box. PMID- 14523561 TI - Distinct but parallel evolutionary patterns between alcohol and aldehyde dehydrogenases: addition of fish/human betaine aldehyde dehydrogenase divergence. AB - Alcohol dehydrogenases (ADHs) of the MDR type (medium-chain dehydrogenases/reductases) have diverged into two evolutionary groups in eukaryotes: a set of 'constant' enzymes (class III) typical of basal enzymes, and a set of 'variable' enzymes (remaining classes) suggesting 'evolving' forms. The variable set has larger overall variability, different segment variability, and variability also in functional segments. Using a major aldehyde dehydrogenase (ALDH) from cod liver and fish ALDHs deduced from the draft genome sequence of Fugu rubripes (Japanese puffer fish), we found that ALDHs form more complex patterns than the ADHs. Nevertheless, ALDHs also group into 'constant' and 'variable' sets, have separate segment variabilities, and distinct functions. Betaine ALDH (class 9 ALDH) is 'constant,' has three segments of variability, all non-functional, and a limited fish/human divergence, reminiscent of the ADH class III pattern. Enzymatic properties of fish betaine ALDH were also determined. Although all ALDH patterns are still not known, overall patterns are related to those of ADH, and group separations may be distinguished. The results can be interpreted functionally, support ALDH isozyme distinctions, and assign properties to the multiplicities of the ADH and ALDH enzymes. PMID- 14523563 TI - Muscle strength testing: evaluation of tests of explosive force production. AB - The purpose of the study was to evaluate four tests of explosive force production (EFP). Specifically, the main aims of the study were to assess the reliability of different EFP tests, to examine their relationship with maximum muscle strength, and to explore the relationship between EFP tests and functional movement performance. After an extensive preliminary familiarization with the tasks, subjects ( n=26) were tested on maximum explosive strength of the elbow extensor and flexor muscle, as well as on rapid elbow extension and flexion movements performed in both an oscillatory and a discrete fashion. In addition to maximum force ( F(max)), four different EFP tests were assessed from the recorded force time curves: the time interval elapsed between achieving 30% and 70% of F(max) ( F(30-70%)), the maximum rate of force development (RFD), the same value normalized with respect to F(max) (RFD/ F(max)), and the force exerted 100 ms after the contraction initiation ( F(100 ms)). Excluding F(30--70%), all remaining EFP tests revealed either good or fair reliability (intraclass correlation coefficients being within 0.8-1 and 0.6-0.8 intervals, respectively) which was also comparable with the reliability of F(max). RFD and F(100 ms) demonstrated a positive relationship with F(max), but not T(30-70%) and RFD/ F(max). Stronger elbow flexor muscles also demonstrated higher values of RFD and F(100 ms) than weaker elbow extensor muscles, while no difference was observed between either T(30-70%) or RFD/ F(max) recorded from two muscles. Despite the simplicity of the tested movement tasks, the relationship observed between the EFP tests and the peak movement velocity remained moderate and partly insignificant. It was concluded that most of the EFP tests could be reliable for assessing neuromuscular function in their muscle-force- (or, indirectly, muscle size) dependent (such as RFD and F(100 ms)), or muscle-force-independent ( T(30 70%) and RFD/ F(max)) forms. However, their "external validity" when applied to assess the ability to perform rapid movements could be questioned. PMID- 14523562 TI - Hormonal responses in athletes: the use of a two bout exercise protocol to detect subtle differences in (over)training status. AB - In overtrained athletes, several signs and symptoms have been associated with the imbalance between training and recovery. However, reliable diagnostic markers for distinguishing between well-trained, overreached (OR) and overtrained (OT) athletes are lacking. A hallmark feature of overtraining syndrome (OTS) is the inability to sustain intense exercise and recover for the next training or competition session. We therefore devised a test protocol utilizing two bouts of maximal work. With this test protocol we tried to establish a difference in hormonal responses between the training status of T and OR athletes. Seven well trained cyclists participated in this study and were tested before and after a training camp. We also present the data of one OT motocross athlete who was clinically diagnosed as overtrained. All athletes performed two maximal exercise tests separated by 4 h. Blood was analyzed for cortisol, adrenocorticotrophic hormone (ACTH), growth hormone and prolactin (PRL). Performance decreased by 6% between the first and the second exercise test in the OR group and by 11% in the OT subject. Moreover, during the second exercise test there were more marked differences between the T and OR athletes; in particular, the OT subject did not show an increase in some of the hormonal responses. PRL increased only by 14% in the OT subject's second test and there was a 7% decrease in ACTH. The two exercise approach enables us to detect subtle performance decrements that will not be identified by one exercise trigger. The hormonal responses to the second exercise test were different between the T and OR athletes (the increase in the T group was higher than in the OR that was higher than in the OT). The results of the case presentation of an overtrained athlete provide evidence of an altered and dysfunctional hypothalamic-pituitary axis response to two bouts of maximal exercise. These findings can be used to develop markers for diagnosis of OTS and to begin to address the pathologic mechanism operative in the syndrome, as well as providing an outcome measure to evaluate possible therapeutic regimes. PMID- 14523564 TI - Effects of shortening velocity and of oxygen consumption on efficiency of contraction in dog gastrocnemius. AB - Previous observations have shown that, in isolated perfused dog gastrocnemii in situ, stimulated to aerobic rhythmic isotonic tetanic contractions (at about 40% of maximal isometric force), only about 20% of the overall metabolic power (proportional to the rate of O2 consumption, VO2) was converted into mechanical power (W). Here we report that, in the same preparation, the maximal velocity during the shortening phase of each tetanus (v, mm s(-1)) increased with the rate of energy dissipation, as given by the difference between VO2 and W (W kg(-1)). The relationship between these variables was described by: v=2.85(VO2-W)(1.24) (R2=0.85; n=17). A mathematical analysis of this equation shows that the overall mechanical efficiency (eta=WVO2(-1)) decreased with increasing v (at constant VO2), whereas it increased with increasing VO2 (at constant v). The net effect of this state of affairs was that the decrease of eta over the entire range of work intensities was relatively minor (from 0.22 to 0.15), in spite of a large increase of v, (from 40 to 120 mm s(-1)), thanks to the concomitant increase of VO2 (from 10 to 25 W kg(-1)). So, under these experimental conditions, the energetics of work performance seems to be governed by two conflicting needs. The need for a sufficiently high shortening speed (and hence power output), itself requiring a sufficiently large energy dissipation rate, which, however, brings about a fall in eta. This is counteracted by the increased VO2, which in turn leads to an increased efficiency at the expense of a fall in shortening speed. PMID- 14523565 TI - Human skeletal muscle energy metabolism: when a physiological model promotes the search for new technologies. AB - The idea that muscle function and, in fact, the function of all living tissues may be described by physical laws appeared in the second half of the 19th century. During this period, Helmoltz (Uber die Erhaltung der Kraft, 1847) showed that the principle of energy conservation may be applied to living systems. In the field of exercise physiology, this idea has been subsequently developed by Rodolfo Margaria's (1901-1983) School and the mathematical formalization of the theory has succeeded in the bioenergetic model. During the last 20 years methodical and critical study of the bioenergetic model has been carried out by Paolo Cerretelli, one of the most important heirs of Rodolfo Margaria's School. Original results and technological developments have been generated by his activity and many young scientists have been educated in this approach. The present paper wants to present the modern history of the bioenergetic model and is dedicated to Paolo Cerretelli on the occasion of his seventieth birthday. PMID- 14523567 TI - Cross section calculations for electron scattering from DNA and RNA bases. AB - Differential and integral cross sections for elastic electron collisions with uracil, cytosine, guanine, adenine and thymine have been calculated using the independent atom method with a static-polarization model potential for incident energies ranging from 50 to 4000 eV. Total cross sections for single electron impact ionization of selected DNA and RNA bases have also been calculated with the binary-encounter-Bethe model from the ionization threshold up to 5000 eV. Cross sections within the investigated energy range, can be related to the molecular symmetry, the number of target electrons and molecular size; elastic and ionization processes are most efficient for guanine and adenine molecules, while the lowest cross sections were obtained for the uracil molecule. The ionization cross sections for cytosine, thymine, adenine and guanine are compared with those recently obtained with a semi-classical and binary-encounter-Bethe formalisms. No theoretical and experimental data for elastic electron scattering from DNA and RNA bases are available, but comparisons with calculations for molecules of similar size and geometry allows the validity of the theoretical approach to be verified. PMID- 14523569 TI - Cerebral vasculitis due to Treponema pallidum infection: MRI and MRA findings. PMID- 14523568 TI - The epidemiology of neonatal tumours. Report of an international working group. AB - Neonatal tumours occur every 12,500-27,500 live births and comprise 2% of childhood malignancies, but there is little clarity as to their real prevalence, sites of origin and pathological nature as reported series vary. As an entity, neonatal tumours provide a unique window of opportunity to study tumours in which minimal environmental interference has occurred. The majority of tumours present with a mass at birth (e.g., teratomas, neuroblastomas, mesoblastic nephroma, fibromatosis), which are not infrequently identified on antenatal ultrasound. Histologically, teratoma and neuroblastoma remain the two main tumour types encountered with soft tissue sarcoma, renal tumours, CNS tumours and leukaemia being the next most common tumour types identified. Malignant tumours are uncommon in the neonatal period per se and benign tumours may have malignant potential. A particular problem exists in clinical classification, as histological features of malignancy do not always correlate with clinical behaviour. Benign tumours may also be life threatening because of their size and location. Other tumours may demonstrate local invasiveness, but no metastatic potential, and tumours that are clearly malignant may demonstrate unpredictable or uncertain behaviour. Screening programmes have brought more tumours to light, but do not appear to affect the overall prognosis. They may provide clues to the stage at which tumours develop in foetu. The aetiology of cancer in children is multifactorial and includes both genetic and environmental factors. The association between congenital abnormalities and tumours is well established (15% of neonatal tumours). Genetic defects are highly likely in neonatal tumours and include those with a high risk of malignancy (e.g., retinoblastoma), but also genetically determined syndromes with an increased risk of malignancy and complex genetic rearrangements. Tumours are mostly genetically related at a cellular level and factors influencing cellular maturation or apoptosis within the developing foetus may continue to operate in the neonatal period. Cytogenetics of neonatal neoplasms appear to differ from neoplasms in older children, thus possibly explaining some of the observed differences in clinical behaviour. Certain constitutional chromosome anomalies, however, specifically favour tumours occurring in the foetal and neonatal period. In support of this hypothesis, certain cytogenetic anomalies appear to be specific to neonates, and a number of examples are explored. Other environmental associations include ionizing radiation, drugs taken during pregnancy, infections, tumours in the mother and environmental exposure. PMID- 14523570 TI - Heart rate variability in patients with rheumatoid arthritis. AB - Heart rate variability (HRV) is a useful tool for the detection of sympathetic parasympathetic balance in the autonomic nervous system. Autonomic nervous system involvement in patients with rheumatoid arthritis (RA) has rarely been studied and has shown conflicting results. Our purpose was to determine if HRV showed changes in patients with RA in comparison with the normal population. Short-term analysis of HRV was performed for time-domain frequency in 42 patients with RA and 44 matched controls. In this analysis, patients displayed lower standard deviation of the mean than healthy subjects ( P<0.0001). Patients tended to display higher pNN50 and root-mean-square of successive difference values than did healthy subjects, but these differences were not statistically significant (P >0.05). In frequency domain analysis, the spectral measures of HRV showed reduced high-frequency (HF) values and an higher low-frequency (LF) values; as a result, the ratio between low and high frequencies (LF/HF), representative of sympathovagal modulation, was significantly increased (P=0.001, P=0.012, and P=0.003, respectively). Our data suggest an increase in sympathetic control of the heart rate in patients with RA. This increased sympathetic activity could play a key role in the development of ventricular tachyarrhythmias in RA and may be related to the higher incidence of sudden death in this disorder. PMID- 14523571 TI - Prevalence of atopy in rheumatoid arthritis in Sivas, Turkey. A prospective clinical study. AB - Assumed mutual antagonism of T-helper cell (Th)1 and Th2 diseases suggests that the prevalence of atopy should be decreased in rheumatoid arthritis (RA). We tried to determine that prevalence. Sixty-two patients with RA and 61 with osteoarthritis (control group) were included in the study. A questionnaire was used concerning mainly the symptoms of atopy. Skin prick tests, pulmonary function tests, chest X-rays, immunoglobulin E levels, and eosinophil counts were obtained. The prevalences of asthma, hay fever, and eczema in the RA group were 3.2%, 14.5%, and 1.6%, respectively. In the control group, they were 6.5%, 22%, and 6.5%, respectively. There was no significant difference between groups. There was also no significant difference between immunoglobulin E levels. Eosinophil counts were higher in the control group, however these values were within the normal range. Skin prick tests were obtained in seven RA patients, and eight controls revealed increased positivity. The prevalence of atopy in rheumatoid arthritis was not different from that in the general population. Our study results cannot support the concept of Th1/Th2 mutual antagonism. PMID- 14523573 TI - STA10 repression of STA gene expression is caused by a defective activator, flo8, in Saccharomyces cerevisiae. AB - The expression of STA genes that encode extracellular glucoamylase isozymes is repressed in most laboratory Saccharomyces cerevisiae strains, which are believed to contain an undefined repressor, designated STA10. To identify the regulator involved in STA10 repression, we investigate the FLO8, MSN1, MSS11, STE12, and TEC1 genes. The Deltaflo8 or Deltamss11 deletion mutants in the sta10 genetic background exhibit both a loss of flocculation ability and a reduction in extracellular glucoamylase activity, as in the STA10 strain. Moreover, the STA10 repression is suppressed completely or partially by the introduction of a single copy of the FLO8 or MSS11 genes. Sequence analysis and complementation testing of the STA10 strain reveal that it has an inactive, mutated flo8-1 allele. A random spore analysis and transplacement (allele replacement) experiment confirms that the repressive phenotype of STA10 is due to the amber mutation of the transcriptional activator, FLO8. PMID- 14523574 TI - A mathematical model for analysis of the cell cycle in cell lines derived from human tumors. AB - The growth of human cancers is characterised by long and variable cell cycle times that are controlled by stochastic events prior to DNA replication and cell division. Treatment with radiotherapy or chemotherapy induces a complex chain of events involving reversible cell cycle arrest and cell death. In this paper we have developed a mathematical model that has the potential to describe the growth of human tumour cells and their responses to therapy. We have used the model to predict the response of cells to mitotic arrest, and have compared the results to experimental data using a human melanoma cell line exposed to the anticancer drug paclitaxel. Cells were analysed for DNA content at multiple time points by flow cytometry. An excellent correspondence was obtained between predicted and experimental data. We discuss possible extensions to the model to describe the behaviour of cell populations in vivo. PMID- 14523575 TI - Exponential mapping of quantitative trait loci governing allometric relationships in organisms. AB - Allometric scaling relationships or quarter-power rules, as a universal biological law, can be viewed as having some genetic component, and the particular genes (or quantitative trait loci, QTL) underlying these allometric relationships can be mapped using molecular markers. We develop a mathematical and statistical model for mapping allometric QTL on the basis of nonlinear power functions using Taylor's approximation theory. Simulation studies indicate that the QTL position and effect can be estimated using our model, but the estimation precision can be improved from the higher- over lower-order approximation when the sample size used and gene effects are small. The application of our approach in a real example from forest trees leads to successful detection of a QTL governing the allometric relationship between 3rd-year stem height and 3rd-year stem biomass. It is expected that our model will have broad implications for genetic, evolutionary, biomedical and breeding research. PMID- 14523576 TI - Numerical approximations of the mean absorption cross-section of a variety of randomly oriented microalgal shapes. AB - The size, shape, and absorption coefficient of a microalgal cell determines, to a first order approximation, the rate at which light is absorbed by the cell. The rate of absorption determines the maximum amount of energy available for photosynthesis, and can be used to calculate the attenuation of light through the water column, including the effect of packaging pigments within discrete particles. In this paper, numerical approximations are made of the mean absorption cross-section of randomly oriented cells, aA. The shapes investigated are spheroids, rectangular prisms with a square base, cylinders, cones and double cones with aspect ratios of 0.25, 0.5, 1, 2, and 4. The results of the numerical simulations are fitted to a modified sigmoid curve, and take advantage of three analytical solutions. The results are presented in a non-dimensionalised format and are independent of size. A simple approximation using a rectangular hyperbolic curve is also given, and an approach for obtaining the upper and lower bounds of aA for more complex shapes is outlined. PMID- 14523577 TI - Coalescent patterns in diploid exchangeable population models. AB - A class of two-sex population models is considered with N females and equal number N of males constituting each generation. Reproduction is assumed to undergo three stages: 1) random mating, 2) exchangeable reproduction, 3) random sex assignment. Treating individuals as pairs of genes at a certain locus we introduce the diploid ancestral process (the past genealogical tree) for n such genes sampled in the current generation. Neither mutation nor selection are assumed. A convergence criterium for the diploid ancestral process is proved as N goes to infinity while n remains unchanged. Conditions are specified when the limiting process (coalescent) is the Kingman coalescent and situations are discussed when the coalescent allows for multiple mergers of ancestral lines. PMID- 14523578 TI - Mutual interactions, potentials, and individual distance in a social aggregation. AB - We formulate a Lagrangian (individual-based) model to investigate the spacing of individuals in a social aggregate (e.g., swarm, flock, school, or herd). Mutual interactions of swarm members have been expressed as the gradient of a potential function in previous theoretical studies. In this specific case, one can construct a Lyapunov function, whose minima correspond to stable stationary states of the system. The range of repulsion (r) and attraction (a) must satisfy r < a for cohesive groups (i.e., short range repulsion and long range attraction). We show quantitatively how repulsion must dominate attraction ( Rr(d+1) > cAa(d+1) where R, A are magnitudes, c is a constant of order 1, and d is the space dimension) to avoid collapse of the group to a tight cluster. We also verify the existence of a well-spaced locally stable state, having a characteristic individual distance. When the number of individuals in a group increases, a dichotomy occurs between swarms in which individual distance is preserved versus those in which the physical size of the group is maintained at the expense of greater crowding. PMID- 14523584 TI - Quantification of nicotinic acetylcholine receptors in human brain using [123I]5 I-A-85380 SPET. AB - The purpose of this study was to assess the utility of a new single-photon emission tomography ligand, [123I]5-iodo-3-[2(S)-2-azetidinylmethoxy]pyridine (5 I-A-85380), to measure regional nAChR binding in human brain. Six healthy nonsmoker subjects (two men and four women, age 33 +/- 15 years) participated in both a bolus (dose: 317 +/- 42 MBq) and a bolus plus constant infusion (dose of bolus: 98 +/- 32 MBq, B/I=6.7 +/- 2.6 h, total dose: 331 +/- 55 MBq) study. The study duration was 5-8 h and 14 h in the former and the latter, respectively. Nonlinear least-squares compartmental analysis was applied to bolus studies to calculate total (VT') and specific (VS') distribution volumes. A two-tissue compartment model was applied to identify VS'. VT' was also calculated in B/I studies. In bolus studies, VT' was well identified by both one- and two-tissue compartment models, with a coefficient of variation of less than 5% in most regions. The two-compartment model gave VT' values of 51, 22, 27, 32, 20, 19, 20, and 17 ml cm(-3) in thalamus, cerebellum, putamen, pons, and frontal, parietal, temporal, and occipital cortices, respectively. The two-compartment model did not identify VS' well. B/I studies provided poor accuracy of VT' measurement, possibly due to deviations from equilibrium conditions. These results demonstrate the feasibility of quantifying high-affinity type nAChRs using [123I]5-I-A-85380 in humans and support the use of VT' measured by bolus studies. PMID- 14523585 TI - The value of SPET/CT in chronic osteomyelitis. AB - Immunoscintigraphy with technetium-99m labelled anti-granulocyte antibodies (AGA) is an equivalent technique to imaging with in vitro-labelled leucocytes, which is now considered state of the art in the diagnostic work-up of patients with suspected post-traumatic chronic osteomyelitis. In this study, we evaluated the use of a combined single-photon emission tomography/computed tomography (SPET/CT) device to improve detection and anatomical definition of inflammatory bone lesions. Twenty-seven patients with 29 sites of suspected bone infection underwent immunoscintigraphy with 750 MBq 99mTc-labelled AGA. Planar scans were acquired immediately, 4 h and 24 h after injection, and combined SPET/CT was performed using a dual-head multifunctional gamma camera equipped with a low power X-ray system. Accumulation of AGA in inflammatory lesions was quantitated, comparing uptake at 4 and 24 h after injection. The validation was based on culture data derived from surgical or biopsy samples (20 lesions in 18 patients) or clinical follow-up without further therapy for more than 6 months (nine lesions). On a lesion-by-lesion basis 19 true positive, one false positive and nine true negative findings were obtained. SPET/CT correctly identified the location of all positive foci in the appendicular skeleton and that of a cold lesion in the axial skeleton. It also enabled differentiation between soft tissue infection, septic arthritis and osteomyelitis, as well as between cortical, corticomedullary and subperiosteal foci. Sensitivity was identical for SPET and SPET/CT (100%), whereas specificity was improved from 78% to 89% by the use of SPET/CT. Combined SPET/CT improves the accuracy of immunoscintigraphy by allowing correct differentiation between soft tissue infection and bone involvement. This technique may gain clinical relevance in the selection of patients for surgical therapy. PMID- 14523572 TI - Nucleo-cytoplasmic partitioning of proteins in plants: implications for the regulation of environmental and developmental signalling. AB - Considerable progress has been made in the past few years in characterising Arabidopsis nuclear transport receptors and in elucidating plant signal transduction pathways that employ nucleo-cytoplasmic partitioning of a member of the signal transduction chain. This review briefly introduces the major principles of nuclear transport of macromolecules across the nuclear envelope and the proteins involved, as they have been described in vertebrates and yeast. Proteins of the plant nuclear transport machinery that have been identified to date are discussed, the focus being on Importin beta-like nuclear transport receptors. Finally, the importance of nucleo-cytoplasmic partitioning as a regulatory tool for signalling is highlighted, and different plant signal transduction pathways that make use of this regulatory potential are presented. PMID- 14523586 TI - Iodine-124-labeled iodo-azomycin-galactoside imaging of tumor hypoxia in mice with serial microPET scanning. AB - Tumor hypoxia, present in many human cancers, can lead to resistance to radiation and chemotherapy, is associated with a more aggressive tumor phenotype and is an independent prognostic factor of clinical outcome. It is therefore important to identify and localize tumor hypoxia in cancer patients. In the current study, serial microPET imaging was used to evaluate iodine-124-labeled iodo-azomycin galactoside ((124)I-IAZG) (4.2-day physical half-life) as a hypoxia imaging agent in 17 MCa breast tumors and six FSaII fibrosarcomas implanted in mice. For comparison, another promising hypoxic-cell PET radiotracer, fluorine-18-labeled fluoro-misonidazole ((18)F-FMISO), was also imaged in the same tumor-bearing animals. Twelve animals were also imaged with (18)F-labeled fluoro-deoxyglucose ((18)F-FDG). In addition, histological examination was performed, and direct measurement of tumor oxygenation status carried out with the Oxylite probe system. Two size groups were used, relatively well-oxygenated tumors in the range of 80-180 mg were designated as small, and those >300 mg and highly hypoxic, as large. Based on the data from 11 MCa and six FSaII tumors, both (124)I-IAZG and (18)F-FMISO images showed high tracer uptake in the large tumors. In (18)F-FMISO images at 1, 3-4, and 6-8 h post-injection (p.i.), there was considerable whole body background activity. In contrast, (124)I-IAZG imaging was optimal when performed at 24-48 h p.i., when the whole-body background had dissipated considerably. As a result, the (124)I-IAZG images at 24-48 h p.i. had higher tumor to whole-body activity contrast than the (18)F-FMISO images at 3-6 h p.i. Region-of-interest analysis was performed as a function of time p.i. and indicated a tumor uptake of 5-10% (of total-body activity) for FMISO at 3-6 h p.i., and of ~17% for IAZG at 48 h p.i. This was corroborated by biodistribution data in that the tumor-to-normal tissue (T/N, normal tissues of blood, heart, lung, liver, spleen, kidney, intestine, and muscle) activity ratios of IAZG at 24 h p.i. was 1.5-2 times higher than those of FMISO at 3 h p.i., with the exception of stomach. Statistical analysis indicated that these differences in T/N ratios were significant. The small tumors were visualized in the (18)F-FDG images, but not in the (124)I-IAZG or (18)F-FMISO images. This was perhaps due to the combined effect of a smaller tumor volume and a lower hypoxic fraction. Oxylite probe measurement indicated a lesser proportion of regions with pO(2)<2.5 mmHg in the small tumors (e.g., pO(2) was <2.5 mmHg in 28% and 67% of the data in small and large FSaII tumors, respectively), and the biodistribution data showed lower uptake of the tracers in the small tumors than in the large tumors. In the first study of its kind, using serial microPET imaging in conjunction with biodistribution analysis and direct probe measurements of local pO(2) to evaluate tumor hypoxia markers, we have provided data showing the potential of (124)I-IAZG for hypoxia imaging. PMID- 14523589 TI - In vivo imaging of rat brain A1 adenosine receptor occupancy by caffeine. PMID- 14523590 TI - The effect of magnetisation transfer contrast on cerebrospinal fluid on motion artefacts on fluid-attenuated inversion-recovery images. AB - We assessed possible advantages of the use of fluid-attenuated inversion-recovery (FLAIR) sequences with magnetisation-transfer contrast (MTC) over conventional FLAIR images. We carried out cranial MRI at 1 tesla on 50 patients with both sequences. In nine patients with multiple sclerosis (MS) we performed a quantitative comparison of the two sequences, looking at the contrast-to-noise ratio between lesions and normal white matter and counting the number of lesions shown using each method. A qualitative comparison on all patients consisted of the analysis of the appearance of the normal parenchyma, of any lesions, and of artefacts, with particular reference to cerebrospinal fluid (CSF) motion artefacts. The quantitative analysis showed no meaningful difference between the two sequences. The cerebral parenchyma and lesions appeared substantially the same with both techniques. With FLAIR MTC there was a clear, and consistent reduction in CSF motion artefacts. FLAIR MTC sequences can usefully be used in place of the conventional sequence at 1 tesla. PMID- 14523591 TI - Treatment of a spontaneous intracranial dissecting aneurysm with stent-assisted coil embolization. AB - Arterial dissection is now recognized as an important cause of stroke. Most reported dissections involve the extracranial portion of the internal carotid artery (ICA) and vertebral arteries. Spontaneous dissecting aneurysms of the intracranial ICA are uncommon. Endovascular treatment for cerebral aneurysms has become widespread; however, the dissecting aneurysm is still difficult to treat if the parent artery has to be preserved. More recently, stenting has been advocated for use with endosaccular coiling, with the coils held in place by the stent. We herein report a spontaneous intracranial ICA dissecting aneurysm in a 51-year-old woman who was treated using a new technique of combined stent and coils. PMID- 14523592 TI - Prescription and non-prescription medicine use in Denmark: association with socio economic position. AB - OBJECTIVE: To analyse the association among different types of medicine use and different measures of socio-economic position (SEP) in one and the same general population. METHODS: Data from The Danish Health and Morbidity Survey 2000 were analysed. The survey was conducted by face-to-face interviews with a representative sample of the adult Danish population (n=16,690). The associations between prescription and over-the-counter (OTC) medicine use and education, occupation and income were assessed by logistic regression analyses. All analyses were adjusted for age, gender and two measures of health status. RESULTS: This cross-sectional analysis of medicine use in a large representative sample of the Danish population found greater use of prescription medicines among disability pensioners and "others" than in salaried employees. Disability pensioners and self-employed individuals used less OTC medicine than salaried employees. Individuals with low income used more prescription medicines but not more OTC medicines, than those with high income. No major differences were found in prescription medicine use with respect to education, but men within the two middle educational groups tended to use prescription medicine less frequently than both lower and higher educated men. A similar trend was not found for women. OTC medicine use was not associated with education for either gender. CONCLUSIONS: The prevalence of prescription medicine use increases with declining SEP, after adjusting for health status. Such an association does not exist for OTC medicine use. The results show that the least affluent have access to prescription medicine. The difference between prescription and OTC medicine use may be explained by a compensation mechanism. PMID- 14523593 TI - Medical management of hypercalcemia. PMID- 14523594 TI - Association between a polymorphism affecting an AP1 binding site in the promoter of the TCIRG1 gene and bone mass in women. AB - The TCIRG1 gene encodes a component of the osteoclast vacuolar proton pump and previous work has shown that inactivating mutations of the TCIRG1 cause autosomal recessive osteopetrosis. In order to determine whether allelic variation in TCIRG1 contributes to the regulation of bone mineral density (BMD) in normal individuals, we studied the relationship between polymorphisms of TCIRG1 and BMD in a population-based cohort of 739 perimenopausal women. Five common polymorphisms were identified: two in the promoter, a conservative change within exon 4, one within intron 4 and one within intron 11. One of the promoter polymorphisms (G-1102A) lay within a consensus recognition site for the AP1 transcription factor. There was a significant association between the G-1102A genotype and BMD at the lumbar spine ( P = 0.01) and femoral neck ( P = 0.03). The association remained significant after correcting for age, weight, height, menopausal status/HRT use and smoking ( P = 0.008 for spine BMD and P = 0.03 for hip BMD), and homozygotes for the -1100 "G" allele had BMD values significantly higher than individuals who carried the -1100 "A" allele at both spine ( P = 0.007) and hip ( P = 0.047). Subgroup analysis showed that the association between G-1102A and BMD was restricted to premenopausal women who comprised 50.6% of the study group. None of the other polymorphisms or haplotypes were significantly associated with BMD in the study group as a whole or in any subgroup. Functional studies will need to be performed to determine the mechanisms that underlie this association, but we conclude that, in this relatively large population, allelic variation at the G-1102A site of TCIRG1 accounts for part of the heritable component of BMD in Scottish women, possibly by affecting peak bone mass. PMID- 14523599 TI - Hypogonadism and hormone replacement therapy on bone mass of adult women with thalassemia major. AB - We studied bone mass and metabolism in 30 adult women (age 28.5 +/- 1.3) with thalassemia major (TM) and evaluated whether prolonged hormone replacement therapy (HRT) was able to optimize bone accrual. TM patients had reduced bone mass, increased bone turnover and lower serum gonadotropin and estradiol levels compared with 10 normal women of similar age. A significant correlation was found between bone mass and sex hormone levels. Six TM patients with normal ovarian function had normal bone turnover markers and modestly low bone mass (lumbar spine -1.29 +/- 0.31; femoral neck -0.60+/-0.21; Z-score). The other 24 TM women were hypogonadic and had significantly lower bone mass for age (lumbar spine 2.35 +/- 0.2, femoral neck -1.83 +/- 0.2) and increased bone turnover relative to eugonadal women. Of the hypogonadal patients, 13 had taken HRT since age 15 +/- 1 years, but their bone mass and turnover markers were not different than untreated hypogonadal patients. In conclusion, while hypogonadism negatively affects bone mass acquisition in adult TM women, HRT at the standard replacement doses is not sufficient to secure optimal bone accrual. PMID- 14523601 TI - Hypertrophy of vascularized bone isograft in rats treated with cyclosporine A. AB - The aim of this study was to investigate the effects of cyclosporine A (CsA) on vascularized tibio-fibula isograft between 12-week-old male Lewis rats. After transplantation, 45 rats were randomly allocated to one of the following 7 treatment groups: (1) 4-week vehicle (n = 5), (2) 4-week CsA (n = 5), (3) 8-week vehicle (n = 10), (4) 8-week CsA (n = 10), (5) 4-week CsA followed by 4-week vehicle (n = 5), (6) 16-week vehicle (n = 5), or (7) 4-week CsA followed by 12 week vehicle (n = 5). In soft X-ray and micro-computed tomography examination, hypertrophic change of the grafted bones was apparent in the 4- and 8-week CsA groups. Mineral apposition rate and bone formation rate of the grafted bones in the 4-week CsA group were markedly higher than those in the 4-week vehicle group. In the 4- and 8-week CsA groups, however, bone mineral density (BMD) of the grafted bones was lower and strength of the reconstructed bones was weaker than the 4- and 8-week vehicle groups. Urinary deoxypyridinoline (DPD) level was higher in the 4- and 8-week CsA groups than in the 4- and 8-week vehicle groups. The group of 4-week CsA followed by 4-week vehicle had a level of urinary DPD equal to the 8-week vehicle group, but their BMD of the grafted bones was lower and strength of the reconstructed bones was weaker than the 8-week vehicle group. By contrast, the group of 4-week CsA followed by 12-week vehicle had BMD of the grafted bones and strength of the reconstructed bones similar to the 16-week vehicle group. These findings demonstrate that short-term CsA treatment induces hypertrophic change of vascularized bone graft with high-turnover bone loss, and strength of the reconstructed bone is gradually restored after the cessation of CsA treatment. PMID- 14523602 TI - Novel aspects on RANK ligand and osteoprotegerin in osteoporosis and vascular disease. AB - The clinical coincidence of osteoporosis and vascular disease has long indicated that common mediators may adversely affect bone metabolism and vascular integrity alike. Receptor activator of NF-kappaB ligand (RANKL) is an important cytokine for bone resorption that acts through its osteoclastic receptor, receptor activator of NF-kappaB (RANK), while osteoprotegerin serves as a decoy receptor that binds RANKL and prevents activation of RANK. Skeletal and vascular cells are sources and targets of RANKL and OPG both in vitro and in vivo. Modulation of the RANKL/RANK/OPG system in animals results in a skeletal and vascular phenotype, and administration of OPG may prevent osteoporosis and vascular calcification. Recent studies on OPG serum levels and gene polymorphisms also suggest an important role of this cytokine system in skeletal and vascular diseases. In summary, there is increasing evidence that RANKL and OPG may link the skeletal with the vascular system. PMID- 14523603 TI - The influence of fluoride exposure on dentin mineralization using an in vitro organ culture model. AB - This study aimed to characterize fluoride-induced alterations in dentin mineralization within a dentin-pulp organ culture system. Tooth sections derived from male Wistar rat incisors were cultured in Trowel-type culture for 14 days, in the presence of 0 mM, 1 mM, 3 mM and 6 mM sodium fluoride. Tooth sections were processed and analyzed for uptake of fluoride, its subsequent effect on dentin mineralization by tetracycline hydrochloride incorporation and mineral composition, expressed as calcium/phosphorous (Ca/P) ratios. Tetracycline hydrochloride incorporation was demonstrated to decrease with increased fluoride exposure, accompanied by significant increases in both Ca/P ratios and fluoride incorporation. These findings provide further evidence that the established alterations in dentin formation during fluorosis are a consequence of disruption to the mineralization process, and provide a model system with which to investigate further the potential role the extracellular matrix plays in inducing the apparent changes in mineral composition. PMID- 14523605 TI - Ultratrace analysis and isotope ratio measurements of long-lived radioisotopes by resonance ionization mass spectrometry (RIMS). AB - Resonance Ionization Mass Spectrometry (RIMS) is a sensitive and selective method for ultratrace analysis of long-lived radioisotopes and isotope ratio measurements. It provides extremely high isobaric suppression and good overall efficiency. The experimental limits of detection are as low as 10(6) atoms per sample and isotopic selectivities of 5x10(12) have been obtained. The widespread potential of RIMS, using different experimental arrangements, is demonstrated for the determination of the radiotoxic isotopes Pu-238 to Pu-244 and Sr-89/Sr-90 in various environmental samples as well as for Ca-41 in nuclear reactor components and biomedical samples. PMID- 14523606 TI - Superheated liquids for extraction of solid residues from winemaking processes. AB - Solid residues from winemaking processes have been subjected to extraction with superheated water-ethanol mixtures. Identification and characterization of the extracted compounds were achieved by spectrophotometry, gas chromatography with either flame-ionization or mass detectors, and-high performance liquid chromatography with UV detection. Extraction was performed statically with single or repeated cycles. All variables affecting the extraction process have been studied and optimised. The extraction time and temperature were 65 min and 210 degrees C, respectively. Extracts comprised two phases--an aqueous phase, rich in phenolic compounds, and an oily phase, comprising mainly fatty acids. The method allows manipulation of extract composition by changing the applied pressure, temperature, water-to-ethanol ratio, and pH. The method is faster than traditional extraction procedures for obtaining valuable compounds from these residues. PMID- 14523607 TI - Influence of chloride and sediment matrix on the extractability of HgS (cinnabar and metacinnabar) by nitric acid. AB - The extractability of metacinnabar and cinnabar, alone or in the presence of some sediment components, with various concentrations of HNO3 (1, 4, 6, and 14 M) was studied. Both forms of HgS (0.2-0.3 mg HgS in 10-20 mL of acid) were insoluble in all HNO3 concentrations as pure compounds. The presence of FeCl3 enhanced solubility of both cinnabar and metacinnabar, especially when concentrated HNO3 was used for the extraction. As the same effect was not obtained in the presence of FeOOH, we concluded that chloride and not Fe3+ was responsible for HgS dissolution. In fact, addition of very low chloride concentration to concentrated HNO3 provoked partial (Cl>10(-4) M) or even total dissolution (Cl>10(-2) M) of HgS. In dilute HNO3 (4-6 M) cinnabar was much less affected by chloride addition than metacinnabar. Extraction of HgS by concentrated HNO3 in the presence of sediment of various salinities demonstrated that the amount of dissolved HgS increased with the increase of the sediment salinity (from freshwater to estuarine and marine sediment), confirming that chloride enhances dissolution of HgS. Removal of chloride by washing the sediment with Milli-Q water significantly reduced dissolution of added HgS during extraction by concentrated HNO3. These results demonstrate that conclusions based on the extraction schemes using concentrated HNO3 as single extractant or as the first extractant in the sequential extraction procedures can be biased. A verification of artifactual oxidation of HgS, when using more concentrated HNO3 as extractant, would help to verify reliability of the applied extraction procedure. PMID- 14523604 TI - Effect of metabotropic glutamate receptor activity on rhythmic discharges of the neonatal rat spinal cord in vitro. AB - To extend our understanding of the network-based properties which enable a neuronal circuit to produce sustained electrical oscillations, we explored the potential contribution of metabotropic glutamate receptors (mGluRs) to generation of rhythmic discharges. The in vitro spinal cord of the neonatal rat was used as a model to find out if electrical patterns characterized by either alternating or synchronous motor pool discharges (recorded from lumbar ventral roots) required mGluR activation or were modulated by it. Alternating patterns of fictive locomotion (induced by NMDA and 5HT) were slowed down and blocked by the broad spectrum mGluR agonist (+/-)-1-aminocyclopentane-trans-1, 3-dicarboxylic acid (t ACPD; 5-50 microM) and unaffected by the broad spectrum mGluR antagonist (RS) alpha-methyl-4-carboxyphenylglycine (MCPG; 1 mM). The regular, synchronous bursting emerging in the presence of strychnine and bicuculline was accelerated by t-ACPD with a commensurate decrease in single burst length, an effect antagonized by MCPG which per se did not affect bursting. The action of t-ACPD was selectively inhibited by the L-type Ca2+ blocker nifedipine which, however, did not change rhythm acceleration evoked by NMDA. These data suggest that neither alternating nor synchronous oscillatory discharges were apparently dependent on mGluR activation via endogenously released glutamate. However, mGluR activation by the agonist t-ACPD modulated rhythmic patterns, indicating that such receptors are a potential target for pharmacological up- or downregulation of spinal rhythmicity. PMID- 14523608 TI - Rapid speciation of Se(IV) and Se(VI) by flow injection-capillary electrophoresis system with contactless conductivity detection. AB - A flow injection-capillary electrophoresis system with contactless conductivity detection and hydrostatic-pressure-generated flow was used for the fast and sensitive speciation of Se(IV) and Se(VI). The sample throughput was 25 samples per hour using a background electrolyte solution containing 8.75 mM L-histidine (His) adjusted to pH 4.00 with acetic acid. The repeatability of peak areas (n=8) was better than 1.41% and the limits of detection were 190 microg L(-1) and 7.5 microg L(-1) for Se(IV) and Se(VI), respectively. The interference from carbonate, typically present in water samples, was eliminated by using a low-pH electrolyte in which carbonate is uncharged and migrates at the EOF front. The method was applied to the analysis of Se(IV) and Se(VI) in soil samples that were spiked with both selenium species and the results for recovery of both selenium species were in good agreement with their introduced concentrations. PMID- 14523609 TI - Gamma-butyrolactone autoregulators and receptor proteins in non- Streptomyces actinomycetes producing commercially important secondary metabolites. AB - The presence of gamma-butyrolactone autoregulators and their receptor proteins were investigated in five representative strains of non- Streptomyces actinomycetes producing commercially important secondary metabolites. Ethyl acetate extracts of culture were assayed using wild-type S. virginiae for virginiae butanolide, S. lavendulae FRI-5 for IM-2, and S. griseus HH1 for A factor. Actinoplanes teichomyceticus and Amycolatopsis mediterranei were shown to produce autoregulators. Corresponding autoregulator-binding activities were found in the crude cell-free lysates of these strains, using the binding assay with tritium-labeled autoregulator analogues as ligands, which suggests that non- Streptomyces actinomycetes might have autoregulator-dependent signaling cascades. PMID- 14523610 TI - Adherence to bisphosphonates and hormone replacement therapy in a tertiary care setting of patients in the CANDOO database. AB - Therapies for osteoporosis must be taken for at least 1 year to be effective. The purpose of this study was to determine the difference in adherence to etidronate, alendronate and hormone replacement therapy in a group of patients seen at our tertiary care centres. The Canadian Database of Osteoporosis and Osteopenia (CANDOO), a prospective observational database designed to capture clinical data, was searched for patients who started therapy following entry into CANDOO. There were 1196 initiating etidronate, 477 alendronate and 294 hormone replacement therapy women and men aged (mean, SD) 65.8 (8.7) years in the study. A Cox proportional hazards regression model was used to assess differences between treatment groups in the time to discontinuation of therapy. Several potential covariates such as anthropometry, medications, illnesses, fractures and lifestyle factors were entered into the model. A forward selection technique was used to generate the final model. Hazard ratios and 95% confidence intervals (CI) were calculated. Adjusted results indicated that alendronate-treated patients were more likely to discontinue therapy as compared with etidronate-treated patients (1.404; 95% CI: 1.150, 1.714). After 1 year, 90.3% of patients were still taking etidronate compared with 77.6% for alendronate. No statistically significant differences were found between hormone replacement therapy and etidronate users (0.971; 95% CI: 0.862, 1.093) and hormone replacement therapy and alendronate users (0.824; 95% CI: 0.624, 1.088) after controlling for potential covariates. After 1 year, 80.1% of patients were still taking hormone replacement therapy, which decreased to 44.5% after 6 years. Increasing age and presence of incident non-vertebral fractures were found to be independent predictors of adherence. In conclusion, alendronate users were more likely to discontinue therapy than etidronate users over the follow-up period. Potential barriers to long-term patient adherence to osteoporosis therapies need to be evaluated. PMID- 14523611 TI - Clinical results of arthroscopic meniscal repair using biodegradable screws. AB - This study was performed to assess the clinical results of meniscus refixation using biodegradable Clearfix meniscal screws. Between July 1999 and June 2001 this technique was performed on 65 patients, of whom 60 (27 women, 33 men; 92%) were followed up by clinical examination after an average of 18 months (range 8 34). The average age of the patients at the time of surgery was 30 years (15-58). Two patients had already undergone a partial meniscectomy at the time of follow up; three patients had had a recurrence of typical clinical signs of a meniscal tear. The other 55 patients rated the overall clinical result as "excellent" (n=21), "good" (n=31) or "satisfactory" (n=3). The average Lysholm score [20] at the time of examination was 93 (49-100) points. The Tegner-Lysholm activity grade [29] preinjury was 5.6 (3-9), compared to a grade of 5.1 (3-9) at the follow-up examination. As a result of our study, biodegradable screws can be recommended as fixation devices with a high rate of good and excellent clinical results. PMID- 14523612 TI - Patients with lateral tracking patella have better pain relief following CT guided tuberosity transfer than patients with unstable patella. AB - In patients with either lateral tracking patella or unstable patella the pathological lateral position of the tuberosity can be corrected by a medial transfer. This study compared the results of subtle CT-guided correction of the tuberosity for objective unstable patella (n=27) with the results for lateral tracking patella (potential instability) as described by Dejour (n=16). Follow-up was 37 months. CT revealed a pathological lateralization of the tibial tuberosity trochlear groove greater than 15 mm in 41 knees. These patients underwent medialization of the tibial tuberosity up to 10-12 mm lateral from the trochlear groove, and 28 patients underwent a distalization to normalize the Caton index to 1.0-1.2. Results were evaluated using Cox' method. Patients with objective patellar instability were rated as 11% excellent, 52% good, 33% fair, and 4% poor. All patients became stable except one who had a 6 degrees valgus alignment. Although 96% had improved stability, 33% of the patients still had pain. The patients with lateral tracking patella (potential instability) were rated as 37.5% excellent, 44% good, and 19% fair. The lower proportion of pain relief in patients with unstable patella is likely the result of the cartilage damage experienced by these patients following multiple dislocations. Thus the patient with lateral tracking patella without patella dislocations must be differentiated from the one with unstable patella. Their prognosis in pain relief is better. PMID- 14523613 TI - In vivo anterior cruciate ligament strain behaviour during a rapid deceleration movement: case report. AB - The mechanism of anterior cruciate ligament (ACL) injury is still unclear. To gain this insight, knowledge of the mechanical behaviour of the healthy ACL during activities that may stress the ligament must be investigated in vivo. The goal of this research was to measure ACL strain in vivo during rapid deceleration, a sport type movement that has been previously shown to precede injuries to the ACL in healthy subjects. A young male subject with no previous knee joint injuries volunteered after informed consent. The strain gauge device (DVRT) was calibrated and surgically implanted in the antero-medial band of the intact ACL. The subject was then transported to the lab for data collection. The zero strain position of the ACL was determined using the slack-taut technique. The subject hopped as quickly as possible from a distance of 1.5 m to the target, an X taped at the centre of a force plate, landing with the instrumented left leg and stopping in the landed position. The entire collection window was five seconds at 1000 Hz. A total of three rapid deceleration trials were collected and averaged over the hop cycle. The slack-taut test was then repeated to ensure proper operation of the DVRT and the reliability of the results. The results showed an average peak strain of the ACL during the instrumented Lachman test of 2.00+/-0.17%. The average peak strain of the ACL during the rapid deceleration task was 5.47+/-0.28%. The data indicate that the RD task caused an increase in peak ACL strain that is much higher than during the instrumented Lachman test, and that the strain begins to increase during the flight phase, prior to landing, and reaches a peak that corresponds to the peak ground reaction force. This technique may be used in further sport-specific movements to gain insight into movement patterns associated with ACL injury mechanisms. PMID- 14523614 TI - Survival and recovery of consciousness in anoxic-ischemic coma after cardiopulmonary resuscitation. AB - OBJECTIVE: In patients who remain unconscious after cardiopulmonary resuscitation (CPR) poor outcome may be predicted with several tests. To use these tests, knowledge of a priori chances of poor outcome after varying periods of unconsciousness is needed. This study is aimed at providing such data. DESIGN AND PATIENTS: Data regarding survival and recovery of consciousness were extracted from registry-based and prospective cohort studies of patients with anoxic ischemic coma. A survival analysis was done using Kaplan-Meier estimates and 28 day outcomes were calculated for all patients unconscious after 24 h and 72 h, and 5 days, 7 days and 14 days after CPR. Patient characteristics and outcomes in our cohort were compared with those of published patient series. RESULTS: After 28 days, 27% of 172 patients from the two cohort studies were alive and conscious, 9% were still unconscious, and 64% had died. The proportion of patients who regained consciousness decreased from 34% of those unconscious within the first 6 h post-CPR to 13% of those still unconscious after 2 weeks. The proportion surviving in an unconscious state increased from 6% of patients who were unconscious initially to 33% of those still unconscious after 2 weeks. The chance of survival remained unchanged up to 7 days after CPR, irrespective of the duration of unconsciousness. Patient characteristics and outcomes in our cohort were comparable to data available from the literature. CONCLUSIONS: The a priori chances of (poor) outcome vary with the duration of unconsciousness after CPR. This study provides data for different time-intervals after CPR. PMID- 14523616 TI - Water self-diffusion tensor changes in an avian genetic developmental model of epilepsy. AB - Diffusion tensor imaging (DTI) was used to investigate whether tissue anisotropy in the developing brain is modified by recurrent seizures in epileptic chickens. Twelve epileptic chickens were sorted equally into two experimental groups at 10 days old. Until the age of 180 days, one group was photically stimulated beginning at an age of 2 weeks and repeated every 2 days while the other group was not stimulated. The photic stimulation induced generalized tonic-clonic seizures, and the unstimulated group did not display seizures. Both treatment groups were imaged at three time points, 45 (juvenile), 90 (adolescent), and 180 (adult) days posthatching, and maps of major and minor elements of anisotropy (eta and epsilon), trace and fractional anisotropy (FA) were generated. The eta, epsilon, and trace values in the hyperstriatum, archistriatum, and optic tectum showed significant changes as a function of developmental time point. Differences and/or interactions due to seizures were seen in the archistriatum and optic tectum for eta, epsilon, and trace with the largest differences between the stimulated and unstimulated birds being seen for eta in juvenile birds in the archistriatum (38.1 x 10(-11) m(2)/s versus 18.0 x 10(-11) m(2)/s) and the optic tectum (53.9 x 10(-11) m(2)/s versus 27.1 x 10(-11) m(2)/s). With the DTI parameters being sensitive to microstructure in the brain, these results demonstrate that seizures produce measurable differences, over unstimulated chickens, in brain structure for juvenile chickens, but the differences disappear as the brain matures. In other words, while seizure activity appears to induce atypical biophysical change (relative to unseizing birds) in the brain at a young age, the change is apparently reversed as the brain matures. PMID- 14523617 TI - Shoot population dynamics of persisting clones of Miscanthus sinensis in the warm temperate region of Japan. AB - The population dynamics of annual shoots in four clonal patches of Miscanthus sinensis was surveyed in terms of clone persistence. Over a study period of 3 years, a stable net shoot number was attained through a balanced replacement of old shoots by tillering 2-3 times a year. The birth rate was significantly correlated with the average monthly temperature. This suggests that a warm climate advances the date of tillerings, and shoots become taller as they emerge earlier. Five cohorts were identified, corresponding to shoots that were born in spring, early summer, summer, autumn and early winter. The autumn cohorts were more numerous and had a greater longevity than the other cohorts; they were thus were best suited for overwintering and, consequently, maintaining the population from one generation to the next. The shoots of earlier cohorts grew too tall to survive the cold of winter, which might cause freezing injury, while later cohorts' shoots excelled in overwintering because of their shortness. However, if the delayed shoots are very short, they will also have difficulty in surviving the early season of growth. Therefore, M. sinensis shoots have an optimum size for survival. PMID- 14523618 TI - Cell-cycle-related variation in proteins in suspension-cultured rice cells. AB - To understand the cell cycle process in plants, we searched for proteins that quantitatively change during the cell cycle in suspension-cultured rice ( Oryza sativa L.) cells. The proteins were analyzed by a two-dimensional polyacrylamide gel electrophoresis image-analysis system. We detected 11 proteins that quantitatively changed during the cell cycle, among which beta-tubulins and a calreticulin-like protein were identified. The amounts of beta-tubulin proteins were low in the M phase and high in the G1 phase. In contrast, mRNAs for two of the three types of beta-tubulin were high in the M phase of the cell cycle. The addition of protease inhibitors MG132 or E64d to the cells decreased the beta tubulin proteins during 24 h, suggesting that beta-tubulin proteins are degraded in vivo by proteases other than those whose activities are inhibited by MG132 or E64d. PMID- 14523620 TI - Fluorescence in situ hybridisation (FISH) analysis of chromosome segregation and interchromosomal effect in spermatozoa of a reciprocal translocation t(9,10)(q11;p11.1) carrier. AB - A couple was referred for exploration of repetitive abortions. The man was found to be a carrier of a balanced reciprocal translocation t(9;10)(q11;p11.1). The meiotic segregation of chromosomes 9 and 10 was analysed in 5,157 spermatozoa from this translocation carrier and in 15,255 spermatozoa from three control donors using three-colour fluorescence in situ hybridisation (FISH). The theoretical viability of the different segregation patterns was performed using the computer system HC Forum developed by the Department of Cytogenetics at the Grenoble University Medical School, La Tronche, France. A normal or balanced constitution was found in 56.25% of the analysed spermatozoa. The tertiary 3:1 segregation mode was the most frequently observed (14.37%). The frequencies of adjacent-1, adjacent-2 and 3:1 interchange modes were 12.85, 9.38 and 7.14% respectively. The cumulative frequency of non-viable imbalance was estimated at 20.91% according to the theorical viability of the different segregation patterns. Spermatozoa aneuploidy frequency was also evaluated for chromosomes X, Y and 18, and there was no evidence of interchromosomal effect in spermatozoa from the translocation carrier. FISH analysis of spermatozoa in combination with the viability theorical estimation of the different segregation patterns could be considered a useful tool for genetic counselling in carriers of reciprocal translocation. PMID- 14523621 TI - Identification of Nepsilon-(carboxymethyl)lysine-positive cells in astroglia-rich primary cultures. AB - Astroglia-rich primary cultures from rat brain were used to investigate the presence in glial cells of Nepsilon-(carboxymethyl)lysine (CML), an advanced glycation endproduct. Westernblot analysis of homogenates of rat brain as well as of astroglia-rich cultures demonstrated the presence of CML-modified proteins in these samples. Immunocytochemical staining of astroglia-rich cultures revealed that only a minority of the cells in these cultures were intensively stained for CML. The staining intensity of CML-positive cells was strongly reduced, if the cells were not permeabilized, indicating that intracellular proteins were CML modified. The CML-positive cells were identified as astrocytes and oligodendrocytes by double-labelling immunocytochemical staining for CML and the cellular markers galactocerebroside, myelin basic protein and glial fibrillary acidic protein. In contrast to other glial cells, microglial cells in astroglia rich cultures were CML-negative. The finding that only a minority of cells in astroglia-rich cultures contains high amounts of intracellular CML-modified proteins indicates that specific properties of these CML-positive cells are responsible for the CML-formation in these cells. PMID- 14523622 TI - Differential expression of cytochrome P450 enzymes in cultured and intact foetal rat ventral mesencephalon. AB - Genetic susceptibility to toxin action leading to nigral cell degeneration in Parkinson's disease (PD) may be dictated by the activity of P450 enzymes in brain. In adult rat brain only CYP2C13/6, CYP2D2, CYP2D5 are found in the substantia nigra. However, little is known concerning the isoforms present in foetal dopaminergic ventral mesencephalon (VM) tissues commonly used to study toxin action. In this investigation, we have determined the expression of P450 enzymes in foetal (VM) slices and in primary cultures of this region. In foetal VM sections immunoreactivity was observed for CYP2C13/6, CYP2D1, CYP2D3, CYP2D4, CYP2D5 and OR. There was no expression of CYP1A2, CYP2B1/2, CYP2C12, CYP2D2 and CYP2E1. In cultured foetal rat VM, immunoreactivity was observed for all P450 enzymes examined, namely CYP1A2, CYP2B1/2, CYP2C13/6, CYP2C12, CYP2D1, CYP2D2, CYP2D3, CYP2D4, CYP2D5, CYP2E1 and OR. There were marked differences in the degree of expression of the isoforms of P450, for example CYP2D1 was only weakly expressed in foetal VM sections but expression was strong in VM cultures. The difference between VM slices and primary cultures suggests that the culturing process can induce some P450 enzymes. CYP2D1, CYP2D3, CYP2D4 were expressed in the foetal VM but were not present in adult rat substantia nigra. Further investigation is now required to determine the functional implications as they may confer an altered level of susceptibility to neurotoxins between foetal and adult dopaminergic cells. PMID- 14523623 TI - Intracellular changes in astrocytes and NG 108-15 neuroblastoma X glioma cells induced by advanced glycation end products. AB - INTRODUCTION: The extracellular effects caused by advanced glycation end products (AGEs) themselves formed by the Maillard-reaction from proteins and sugars, are well known. Less is known about the effects of AGEs on cellular function. Cellular dysfunctions accumulate with increasing age and can be found as a pathogenetic principle of different diseases where AGEs are involved. The causal mechanisms of these cellular dysfunctions are not fully understood. MATERIAL AND METHODS: AGEs effects were examined in cell culture using video microscopy, immunohistochemistry, flow cytometry and biochemical methods in neural (NG 108 15) and glial (primary culture of astrocytes) cells. RESULTS: AGEs can bind to the cell surface of neural and glial cells. AGEs induce an altered distribution of vesicles and a statistically significant reduction in the velocity of microvesicles. The proteins dynamin 2 and clathrin, which are an important part of the transport apparatus, show changes in intracellular distribution similar to that of microvesicles. The immunoreactivity of dynamin 2 in AGE-treated cells is increased after AGE-incubation. After incubation durations of 2 h, 6 h and 12 h we found an increase (maximum +34.6%) of the relative fluorescence intensities of dynamin 2 in the AGE-BSA treated cells in comparison to the control cells (BSA). Using flow cytometry we found a minimal increase (14.4%) in clathrin immunoreactivity after 2 h incubation with 500 nM AGE-BSA. The non-quantitative western blot showed no difference of the electrophoretic behaviour of both dynamin 2 and clathrin in AGE-BSA-incubated cells (2 h, 500 nM) and in control cells (2 h, 500 nM BSA-incubation). The incubation with AGE-BSA up to a concentration of 500 nM in culture was not followed by apoptotic or necrotic cell death. DISCUSSION: We could show defined effects on the cells induced by incubation with AGEs. One of the effects is a significant reduction of the velocity of microvesicles, the reason for this is unclear. It could be due to disturbed calcium metabolism or caused by microtubuli-changes. The aggregation of intracellular microvesicles could be the result of an altered binding-behaviour of the vesicles or the destruction of the transport apparatus. The proteins dynamin 2 and clathrin, which are involved in transport apparatus, show changes in intracellular distribution indicating a breakdown of the normal cellular distribution system. The current results may contribute to the understanding of the pathologic processes in the cell in Mb. Alzheimer and other neurodegenerative diseases, where AGEs-accumulation can be found in tissues. PMID- 14523624 TI - Dopamine receptor agonists in current clinical use: comparative dopamine receptor binding profiles defined in the human striatum. AB - The aim of this study was to compare dopamine receptor binding affinities of all currently approved dopamine receptor agonist treatments for Parkinson's disease (PD) in human brain tissue. Alpha-dihydroergocryptine and lisuride displayed higher comparative affinities (Ki=35.4 and 56.7 nM, respectively) for D1 receptors, than the D1/D2 dopamine agonist pergolide (Ki=447 nM). The second generation non-ergot dopamine receptors agonists pramipexole and ropinirole demonstrated no affinity for D1 receptors at concentrations up to 10(-4) M. The ergoline dopamine agonists cabergoline and lisuride displayed the highest affinities for the D2 receptor (Ki=0.61 and 0.95 nM, respectively). Surprisingly, the second generation non-ergot dopamine receptors agonists pramipexole and ropinirole only weakly inhibited binding to D2 receptors (Ki=79.5 and 98.7 microM, respectively using [3H]spiperone). Interestingly we also found that the affinities of cabergoline (Ki=1.27 nM), lisuride (Ki=1.08 nM) and pergolide (Ki=0.86 nM) for the D3 receptor subtype were comparable to that of pramipexole (Ki=0.97 nM). The present results thus support the hypothesis that the antiparkinsonian effect of dopamine receptor agonists is mediated by a more complex interactions with dopamine receptor subtypes than currently believed. PMID- 14523626 TI - Apraxia of eyelid opening in a case of atypical corticobasal degeneration. AB - Apraxia of eyelid opening (AEO) occurs in several clinical conditions, even in the absence of any other neurological sign; nonetheless, in most of the cases AEO has been reported in association with basal ganglia diseases, such as corticobasal degeneration (CBD). We describe a patient with a clinical diagnosis of frontotemporal dementia who, later, developed parkinsonian signs and AEO. We suggest that the finding of AEO in patients with a frontotemporal syndrome could be a helpful expedient for the early diagnosis of atypical clinical findings of CBD, characterised by behavioural and cognitive aspects at first. PMID- 14523625 TI - Cerebral alterations in a MPTP-mouse model of Parkinson's disease--an immunocytochemical study. AB - We investigated the immunohistochemical alterations of neuronal nitric oxide synthase (nNOS), endothelial NOS (eNOS), tyrosine hydroxylase (TH), microtubule associated protein 2a,b (MAP 2), glial fibrillary acidic protein (GFAP), parvalbumin (PV), and dopamine transporter (DAT) in the striatum and substantia nigra following the application of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in mice. TH-, MAP 2- and DAT-immunoreactive cells were decreased gradually in the striatum and substantia nigra from 1 day up to 7 days after MPTP treatment, as well as the reduction of the striatal dopamine, DOPAC and HVA content. The number of GFAP-immunoreactive astrocytes increased gradually in the striatum and substantia nigra from 1 day up to 7 days after MPTP treatment. Striatal nNOS-immunoreactive cells were unchanged in MPTP-treated mice. In the substantia nigra, intense immunoreactivity of nNOS-positive cells increased 5 hr after MPTP treatment. Thereafter, the immunoreactivity of nNOS-positive cells decreased gradually from 1 day up to 7 days after MPTP treatment. eNOS immunopositive cells were unchanged in the striatum and substantia nigra. These results demonstrate that nNOS may play a key role in the development of MPTP neurotoxicity. Our findings also indicate that MPTP can cause the functional damage of interneurons in the substantia nigra, but not in the striatum. PMID- 14523627 TI - CSF-tau, CSF-Abeta1-42, ApoE-genotype and clinical parameters in the diagnosis of Alzheimer's disease: combination of CSF-tau and MMSE yields highest sensitivity and specificity. AB - This study evaluated the sensitivity and specificity of the cerebrospinal fluid (CSF) levels of tau-protein, amyloid-beta-peptide 1-42 (Abeta1-42), ApoE-genotype and the degree of cognitive decline as diagnostic markers for Alzheimer's disease (AD). Data was obtained from 105 AD patients and 68 controls. Median CSF-tau levels were increased (512 pg/ml vs. 145 pg/ml, p<0.001) and Abeta1-42-levels were decreased (238.5 pg/ml vs. 310 pg/ml, p<0.001) in AD patients compared to controls. A weak correlation was found between CSF-Abeta1-42 and MMSE score (r=.245). Within all subjects, a correlation of CSF-Abeta1-42 (r=-.337) and CSF tau (r=.384) with age was found. The combination of CSF-tau levels and MMSE revealed the highest sensitivity (92%) and specificity (87%). In summary, CSF-tau was a useful biological marker to discriminate AD from normal aging, neurological and psychiatric disorders. CSF-Abeta1-42 showed no additional benefit in discriminating patients from controls but might be useful for tracking the severity of the disease. PMID- 14523628 TI - Distribution of anxiogenic-induced c-Fos in the forebrain regions of developing rats. AB - An anxiogenic or a pharmacological stressor, N-methyl-beta-carboline-3 carboxamide (FG-7142), (20 mg/kg, intraperitoneally injected) induced a dense nuclear c-Fos-like immunoreactivity in the pyriform cortex, cingulate and retrosplenial cortex, layers II-VI of the neocortex, lateral habenula, lateral septum, paraventricular nucleus of the thalamus, striatum, central and medial nucleus of the amygdala, but a sparse c-Fos immunostaining in the hippocampus and layer I of the neocortex in the forebrain of 56-day-old rats. Among these regions, the 8-day-old rats expressed much fewer c-Fos-positive cells in the neocortex, lateral habenula, lateral septum and medial nucleus of the amygdala than the young adult rats following the FG-7142 injection. These differences in the regional distribution of a neuronal activity marker, c-Fos, could reflect the postnatal development of neuronal populations or neuron circuits involved in stress and/or emotional response in the forebrain. PMID- 14523629 TI - Specific modulation of sigma binding sites by the anxiolytic drug opipramol. AB - The atypical anxiolytic and antidepressive drug opipramol binds with high affinity to sigma1 and somewhat lower affinity to sigma2 sites. After subchronic treatment, opipramol significantly down-regulated sigma2 but not sigma1 sites. This effect was not seen for imipramine, citalopram, and reboxetine under similar conditions. On the other hand, only imipramine reduced the number of sigma1 sites. It is suggested that effects at sigma2 sites are involved in the anxiolytic properties of opipramol. PMID- 14523630 TI - Autonomic responses in boys with externalizing disorders. AB - OBJECTIVE: Autonomous hyporesponsiveness in children appears to be an indicator of a greater future likelihood of antisocial behavior. Since externalizing disorders are suggested to implicate a risk of antisocial behavior in later life, psychophysiological measurements were assessed commonly used in the realm of antisocial behavior. METHOD: Arousal measures and electrodermal responses to orienting and startling stimuli were assessed in 8-13 year old boys diagnosed with attention deficit/hyperactivity disorder (ADHD), conduct disorder (CD), the comorbid condition of ADHD+CD, and in controls. The eyeblink response to startling stimuli was also recorded. RESULTS: Children with CD and ADHD+CD showed decreased electrodermal responses and accelerated habituation in all paradigms compared to children with ADHD alone and controls. CONCLUSION: CD boys with and without ADHD showed a widely analogous response pattern which is highly similar to that found in antisocial adults. Data suggest that it is CD that sharply rises the risk of unfavorable, antisocial development, whether it occurs alone or comorbid with ADHD. PMID- 14523631 TI - Arbuscular mycorrhizal fungi associated with common pteridophytes in Dujiangyan, southwest China. AB - The colonization and diversity of arbuscular mycorrhizal (AM) fungi associated with common pteridophytes were investigated in Dujiangyan, southwest China. Of the 34 species of ferns from 16 families collected, 31 were colonized by AM fungi. The mean percentage root length colonized was 15%, ranging from 0 to 47%. Nineteen species formed Paris-type and 10 intermediate-type AM. In two ferns, only rare intercellular non-septate hyphae or vesicles were observed in the roots and AM type could not be determined. Of the 40 AM fungal taxa belonging to five genera isolated from rooting-zone soils, 32 belonged to Glomus, five to Acaulospora, one to Archaeospora, one to Entrophospora, and one to Gigaspora. Acaulospora and Glomus were the dominant genera and Glomus versiforme was the most common species. The average AM spore density was 213 per 100 g air-dried soil and the average species richness was 3.7 AM species per soil sample. There was no correlation between spore density and percentage root length colonized by AM fungi. PMID- 14523632 TI - Care of breast cancer in the elderly woman--what does comprehensive geriatric assessment (CGA) help? AB - Breast cancer is the most common cancer in women in Europe and Northern America. Its incidence and mortality rates are increasing with advance in age. Only few elderly women with breast cancer are treated in clinical trials. Elderly women with the same numerical age are very heterogeneous considering their biological age. Geriatric medicine has established comprehensive geriatric assessment (CGA) to get important information on elderly patients missed by a routine clinical history and physical examination. The data collected in CGA are of prognostic relevance for mortality, morbidity, maintenance of independence and utilisation of health care resources. Within the last few years, some research groups have demonstrated that the use of CGA in elderly cancer patients collects information missed by the currently established workup of these patients and that this information is of prognostic relevance concerning toxicity of chemotherapy and mortality. The use of CGA in a population of general elderly patients improves functional status and mental health, but so far no effect on mortality could be demonstrated. A subgroup analysis of the elderly cancer patients within this trial additionally demonstrated an improvement in pain control. Special data concerning the use of CGA within the group of elderly breast cancer patients are not published so far. The authors suggest areas of care for elderly women with breast cancer within which the CGA might be able to improve treatment and which should be a field of randomised controlled trials in the future. PMID- 14523633 TI - Preterm neonates with nephrocalcinosis: natural course and renal function. AB - The aim of the study was to evaluate the natural course of nephrocalcinosis (NC) in preterm neonates and the effect of NC on blood pressure and renal glomerular and tubular function. In a prospective observational study of 201 preterm neonates (gestational age <32 weeks) NC was present at term in 83 patients (41%), who were subsequently examined at 6, 12, and 24 months, and until August 2000 annually (with a maximum of 4 years) if NC persisted. Examination consisted of blood pressure measurement, renal ultrasonography, and glomerular and tubular function tests. The probability that NC, when present at term, would persist for 15 and 30 months was 34% [21-45, 95% confidence interval (CI)] and 15% (5-25, 95% CI) (Kaplan-Meier), respectively. Urinary tract infection did not occur more frequently in patients with NC (2.5%) than patients without NC at term (4.4%). Systolic and diastolic blood pressures above the 95th percentile were found in 39% and 48% of patients at 1 year and 30% and 34% at 2 years ( P<0.001). Mean glomerular filtration rate (GFR) (inulin clearance) at 1 and 2 years was 92 and 102 ml/min per 1.73 m(2), respectively. TP/GFR and excretion of alpha(1) microglobulin were normal. The desmopressin test was impaired in 4 of 30 patients at 1 year and 2 of 25 at 2 years. It was concluded that while proximal tubular function is unaffected in children with neonatal NC, high blood pressure and impaired glomerular and distal tubular function might occur more frequently than in healthy children. Although no relationship can be proven between NC and hypertension or diminished renal function in this study, these results justify a large follow-up study with matched controlled study groups. PMID- 14523634 TI - New renal scars in children with severe VUR: a 10-year study of randomized treatment. AB - The International Reflux Study in Children was set up to compare prospectively the outcome of medical or surgical management of children with grade III or IV vesicoureteral reflux and a history of symptomatic urinary tract infection. Development of new radiological scars was the main end point. Of the 306 children randomized, 302 (153 medical, 149 surgical) were available for radiological follow-up at 5 years. New scars had developed in 19 medically and 21 surgically treated children. Among 223 patients (113 medical, 110 surgical) who continued follow-up with urography at 10 years, only 2 further new scars developed. Overall, 47 new scars were acquired in 42 patients (20 medical, 22 surgical), 25 of them in children with unscarred kidneys at entry (13 medical, 12 surgical). New scars occurred mostly in children under 5 years of age and were observed more frequently in children with grade IV than grade III reflux. We conclude that with careful management, only a small proportion of children with severe reflux developed new scars and rarely after the first 5-year follow up period, and that there was no difference between children treated medically or surgically. PMID- 14523635 TI - Pre dialysis of blood prime in continuous hemodialysis normalizes pH and electrolytes. AB - In critically ill children weighing <10 kg, it is necessary to use blood as a priming solution for the extracorporeal continuous renal replacement therapy (CRRT) circuit before initiating CRRT to prevent hemodilution and maintain adequate oxygenation. However, blood bank blood usually contains supra physiological electrolyte concentrations and a non-physiological acid-base balance that may exacerbate the patient's condition. The objective of this trial was to develop a simple protocol to pre-treat blood bank-derived blood to yield a more physiological blood priming solution. Expired human blood in a recirculating in vitro CRRT circuit was dialyzed prior to the initiation of CRRT using a physiological dialysate solution. Serial blood samples were assessed for electrolyte and pH content. Regimens using maximal blood flow rates (180-200 ml/min) and aggressive dialysate flow rates (33-42 ml/min) were able to correct severely hyperkalemic and acidemic blood within 7.5 min. Initially elevated blood potassium concentrations >20 mEq/l were normalized to below 5 mEq/l within 7.5 min of dialysis in all cases. Blood bank-derived blood can be "conditioned" quickly to physiological pH and electrolyte concentrations using these simple pre dialysis regimens. Unlike some blood preparation regimens that have been published, the technique used in this trial requires no special equipment or added medications that are not already used in CRRT. PMID- 14523636 TI - Urinary excretion of endothelin-1 in children with absorptive idiopathic hypercalciuria. AB - Urinary excretion of endothelin-1 (ET-1) and plasma ET-1 were measured in 21 children with absorptive idiopathic hypercalciuria (AIH) and 22 controls. The absorptive type of idiopathic hypercalciuria was determined by a calcium loading test. Daily urinary excretion of ET-1 and urinary ET-1/creatinine ratio were significantly increased ( P=0.005 and P=0.007, respectively) in patients with AIH (9274+/-6444 pg/24 h and 14.04+/-9.52 pg/mg, respectively) compared with controls (4699+/-2120 pg/24 h and 7.36+/-4.71 pg/mg, respectively). Plasma ET-1 levels were significantly lower in patients with AIH (0.84+/-0.64 pg/ml) than in controls (1.54+/-0.54 pg/ml, P=0.0001). In conclusion, patients with AIH had increased urinary ET-1 excretion and decreased plasma ET-1 levels. This is most likely due to the decreased reabsorption of ET-1 in the renal tubule and increased renal production. PMID- 14523637 TI - Carnitine supplementation improves apolipoprotein B levels in pediatric peritoneal dialysis patients. AB - There have been conflicting reports concerning the effect of carnitine supplementation on lipid metabolism in patients on peritoneal dialysis (PD). We investigated several parameters of lipid metabolism in pediatric PD patients supplemented with carnitine. The study included 20 patients receiving PD (treatment group) aged 2-18 years and a matched healthy control group. In the treatment group, baseline triglyceride, total cholesterol, low-density lipoprotein cholesterol, very low-density lipoprotein cholesterol, and apolipoprotein B levels were higher than in the control group. High-density lipoprotein cholesterol, free fatty acid, phospholipids, and apolipoprotein A-I levels were not different from those in the control group. The baseline plasma free carnitine level was lower and acyl-carnitine level was higher in the treatment group. No difference was found between the groups with respect to plasma total carnitine levels. Oral l-carnitine supplementation (50 mg/kg per day for 30 days) led to a significant decrease (from a baseline value of 146.6+/-51.8 mg/dl to 63.6+/-22.2 mg/dl, P<0.001) in apolipoprotein B levels, and no significant change in the other lipid parameters of the treatment group. Oral l carnitine supplementation does not ameliorate the lipid profile in pediatric PD patients, but it causes a significant decrease in apolipoprotein B levels. Hence, carnitine supplementation may be recommended for decreasing apolipoprotein B levels in this patient population. PMID- 14523639 TI - Age variation in a fluctuating population of the common vole. AB - We analysed variation in age in a fluctuating population of the common vole ( Microtus arvalis) in southern Moravia, Czech Republic, to test the assumption of the senescence hypothesis that the age of voles increases with increasing population density. Between 1996 and 1998, we monitored the demographic changes by snap-trapping and live-trapping in a field population passing through the increase, peak and decline phase of the population cycle. We used the eye lens mass method to determine the age of snap-trapped animals and those that died in live-traps. The average age of winter males was clearly higher after the peak phase breeding season than before it. No such phase-dependent shift in age, however, was observed in the female component. Male age continued to increase from autumn to spring over the pre-peak winter, and the highest age was in spring of the peak phase year. However, after the peak phase breeding season the highest age was achieved in winter, with the decline phase males during the next spring tending to be younger. The average age of females in spring populations was always lower than in winter populations. The average age of voles from live-traps was always higher than voles from snap-traps, particularly in winter and spring populations, suggesting the presence of senescent animals. Although the density dependent changes in age are consistent with those observed for other voles, they provide only weak evidence that population cycles in the common vole are accompanied by pronounced shifts in individual age, particularly in female voles. PMID- 14523640 TI - Distribution of MAP1A, MAP1B, and MAP2A&B during layer formation in the optic tectum of developing chick embryos. AB - The expression patterns of three microtubule-associated proteins (MAP1A, MAP1B, and MAP2A&B) were investigated in the developing optic tectum. Expression of MAP1B and middle-molecular-weight peptide of neurofilament (NF-M) was first observed in the same mesencephalic cells on day 3 of incubation, indicating that neuroblasts had been produced. At day 5, MAP1A and MAP2A&B expression appeared in the cellular layer containing the first neuroblasts that differentiate into large multipolar cells. The NF-M+ neurites in the striatum album centrale (SAC) and the striatum opticum (SO) were MAP1B+ up to day 19, but the intensity of MAP1B immunoreactivity decreased with development. All three MAPs were expressed in large multipolar neurons in the developing stratum griseum centrale from the beginning of maturation. Stratum griseum et fibrosum centrale cellular layers, containing radially arranged piriform neurons, were MAP1A-/MAP2A&B- on day 11 but became MAP1A+/MAP2A&B+ during later stages. These results suggest that the timing of MAP expression in neuronal maturation of large multipolar cells differs from that of piriform cells. The expression of MAPs has revealed specific cellular events in the developing optic tectum. Based on our observations, the development of the optic tectum can be divided into four periods. PMID- 14523638 TI - Simplified treatment strategies to fluid therapy in diarrhea. AB - Dehydration resulting from diarrhea remains an important cause of morbidity and mortality among infants and children worldwide. Although it is well established that rapid and generous intravenous restoration of extracellular fluid, followed by oral rehydration therapy (ORT) should be used in children with severe dehydration, physicians continue to be reluctant to use such therapy. Applying the principle of body fluid physiology to the current treatment of dehydration, we developed a simple and yet effective treatment strategy to fluid therapy for children with diarrheal dehydration using commercially manufactured solutions. Children with mild-to-moderate dehydration are best treated with ORT using commercially available oral solutions containing 45-75 mEq/l of Na(+). Children who have clinical evidence of severe dehydration should receive intravenous fluids, 60-100 ml/kg of 0.9% saline in the first 2-4 h to restore circulation. Oliguric patients with severe acidosis should receive a physiological dose of bicarbonate to correct blood pH level to 7.25. Once circulation is restored, the ORT should be given in small quantities to replace losses of water and Na(+) over 6-8 h. Age-appropriate diet should be started as soon as tolerated. Those who cannot tolerate ORT should receive intravenous rehydration for the remainder of the deficit and maintenance. Addition of 20 mEq/l K(+) to rehydration solutions permits repair of cellular K(+ )deficits without risk of hyperkalemia. The amount of Na(+) given to replace maintenance and deficit fluids varies with the forms of dehydration. Isonatremic dehydration is best treated with 5% dextrose in 0.45% saline containing 20 mEq/l KCl over 24 h. Hyponatremic dehydration is best treated with 0.9% saline and 0.45% saline alternately in a 1:1 ratio in 5% dextrose containing 20 mEq/l KCl over 24 h. Hypernatremic dehydration is best treated with 5% dextrose in 0.2% saline containing 20 mEq/l KCl over 2-3 days to avoid cerebral edema. Maintenance hydration is best treated with 5% dextrose in 0.2% saline containing 20 mEq/l KCl. Ideal commercial intravenous maintenance and deficit solutions have yet to appear. PMID- 14523642 TI - Adenylyl cyclase isoforms in rat testis and spermatozoa from the cauda epididymidis. AB - Expression of adenylyl cyclase genes in rat testis and spermatozoa from the cauda epididymidis was investigated using RT-PCR analysis. Genes encoding the transmembrane adenylyl cyclases (tmAC) II, III, IV, V, VI, VII, and VIII were expressed in the testis, whereas only the gene for tmAC III was expressed in caudal spermatozoa. Immunocytochemistry was used to investigate which tmAC were translated into putative, functional proteins in spermatozoa. Indirect immunofluorescence localized the tmAC II enzyme to a region on the head occupied by the acrosome. The tmAC III enzyme was localized to the posterior margin of the head and to the flagellum, whereas tmAC V and/or VI was localized to the region where the ventral surface of the acrosomal equatorial segment is located. The tmAC VII and VIII enzymes were localized to the convex margin of the head, covering the dorsal region of the acrosomal crescent. To our knowledge, this is the first demonstration that five apparently different tmAC enzymes are localized to discrete subcellular regions of mammalian spermatozoa. These findings provide a fundamental basis for future studies, to determine the physiological roles of tmAC in testis and mature spermatozoa. PMID- 14523643 TI - Spatiotemporal distribution of insulin-like growth factor receptors during nephrogenesis in fetuses from normal and diabetic rats. AB - Exposure to hyperglycemia in utero impairs rat nephrogenesis. The effect of maternal diabetes on insulin-like growth factors and their receptors in the fetal kidney is associated with an increase in both mRNA and protein of the insulin like growth factor II/mannose 6-phosphate receptor. However, this receptor has never been localized in the fetal kidney. The spatial and temporal distribution of the three insulin-like growth factor receptors (insulin-like growth factor I receptor, insulin-like growth factor II/mannose 6-phosphate receptor and insulin receptor) in rat metanephros during both normal and streptozotocin-induced diabetic renal development was investigated using in situ hybridization and immunohistochemistry. All receptors were found in the fetal kidney from the start of nephrogenesis. Insulin-like growth factor I receptor expression was ubiquitous and continuously present during metanephric development. Insulin receptor expression was developmentally regulated during kidney maturation with an enhanced expression in proximal tubules at the late stages of development. Insulin-like growth factor II/mannose 6-phosphate receptor expression was ubiquitous in the early stages of development and was dramatically decreased at the late stages of normal kidney development. Insulin receptor and insulin-like growth factor I receptor expressions were unchanged in diabetic metanephroi. Although the spatial expression of insulin-like growth factor II/mannose 6 phosphate receptor was unaffected by hyperglycemia, its expression was not downregulated in the mesenchyme of the nephrogenic zone of diabetic fetuses on gestational day 20. This study suggests a crucial role of insulin-like growth factor II/mannose 6-phosphate receptor in the pathogenesis of the impaired nephrogenesis in fetuses of diabetic mothers. PMID- 14523644 TI - Immunohistochemical analysis of intracardiac ganglia of the rat heart. AB - The neurochemistry of intracardiac neurons in whole-mount preparations of the intrinsic ganglia was investigated. This technique allowed the study of the morphology of the ganglionated nerve plexus found within the atria as well as of individual neurons. Intracardiac ganglia formed a ring-like plexus around the entry of the pulmonary veins and were interconnected by a series of fine nerve fibres. All intracardiac neurons contained immunoreactivity to PGP-9.5, choline acetyl transferase (ChAT) and neuropeptide Y (NPY). Two smaller subpopulations were immunoreactive to calbindin or nitric oxide synthase. Furthermore, a subpopulation (approximately 6%) of PGP-9.5/ChAT/NPY-immunoreactive cells lacking both calbindin and nitric oxide synthase (NOS) was surrounded by pericellular baskets immunoreactive to ChAT and calbindin. Vasoactive intestinal peptide (VIP), calcitonin gene-related peptide (CGRP), pituitary adenylate cyclase activated peptide (PACAP), substance P and tyrosine hydroxylase (TH) immunoreactivity was observed in nerve fibres within the ganglion, but never in neuronal somata. Furthermore, immunoreactivity for NPY was not observed in pericellular baskets surrounding intracardiac neurons, despite being present in all intrinsic neuronal cell bodies. Taken together, the results of this study indicate a moderate level of chemical diversity within the intracardiac neurons of the rat. Such chemical diversity may reflect functional specialisation of neurons in the intracardiac ganglia. PMID- 14523646 TI - Hindsight judgement on ambiguous episodes of suspected infection in critically ill children: poor consensus amongst experts? AB - Few episodes of suspected infection observed in paediatric intensive care are classifiable without ambiguity by a priori defined criteria. Most require additional expert judgement. Recently, we observed a high variability in antibiotic prescription rates, not explained by the patients' clinical data or underlying diseases. We hypothesised that the disagreement of experts in adjudication of episodes of suspected infection could be one of the potential causes for this variability. During a 5-month period, we included all patients of a 19-bed multidisciplinary, tertiary, neonatal and paediatric intensive care unit, in whom infection was clinically suspected and antibiotics were prescribed ( n=183). Three experts (two senior ICU physicians and a specialist in infectious diseases) were provided with all patient data, laboratory and microbiological findings. All experts classified episodes according to a priori defined criteria into: proven sepsis, probable sepsis (negative cultures), localised infection and no infection. Episodes of proven viral infection and incomplete data sets were excluded. Of the remaining 167 episodes, 48 were classifiable by a priori criteria ( n=28 proven sepsis, n= 20 no infection). The three experts only achieved limited agreement beyond chance in the remaining 119 episodes (kappa = 0.32, and kappa = 0.19 amongst the ICU physicians). The kappa is a measure of the degree of agreement beyond what would be expected by chance alone, with 0 indicating the chance result and 1 indicating perfect agreement. CONCLUSION: agreement of specialists in hindsight adjudication of episodes of suspected infection is of questionable reliability. PMID- 14523645 TI - Identification of plant-regulated genes in Ustilago maydis by enhancer-trapping mutagenesis. AB - To identify plant-induced genes in the maize pathogenic fungus Ustilago maydis we have developed a genetic screen that combines REMI (restriction enzyme mediated integration) mutagenesis with enhancer trapping using the gene for Green Fluorescent Protein (GFP) as vital reporter. Of 2,350 insertion mutants isolated, three were shown to express GFP only after the fungus had come into contact with the host maize plant. One of the genes tagged was mfa1, which encodes the pheromone precursor, while the second gene, pig2, codes for a product that showed similarity to protein disulfide isomerase. The third integration event had occurred in a locus which we designated the p -locus. This locus contains 11 genes in a 24-kb stretch. Of these, pig3, 4, 5, 6 and 7 show a plant-regulated expression pattern, while the other genes found at the locus (designated npi) do not. Of the plant-regulated genes only two were found to be similar to database entries: the pig4 product is related to membrane transporters of the major facilitator family, while the pig6 protein shows similarity to multidrug transporters. Detailed expression studies revealed that the five plant-regulated genes at the p -locus differ in their expression profiles. Mutants deleted for each of them showed no apparent phenotype, while the npi1 gene appeared to be essential. A viable deletion encompassing the entire p -locus could be generated when npi1 function was provided ectopically. This deletion mutant also showed no obvious alteration in virulence. PMID- 14523648 TI - Analysis of the embryonic phenotype of Bent tail, a mouse model for X-linked neural tube defects. AB - Neural tube defects, mostly believed to result from closure defects of the neural tube during embryonic development, are frequently observed congenital malformations in humans. Since the etiology of these defects is not well understood yet, many animal models for neural tube defects, either arising from spontaneous mutations or generated by gene targeting, are being studied. The Bent tail mouse is a model for X-linked neural tube defects. This mutant has a characteristic short and kinked tail. Exencephaly occurs in Bent tail embryos with a frequency of 11-16%. Laterality defects also belong to the phenotypic spectrum. In this study, we analyzed the embryonic phenotype in further detail using scanning electron microscopy during the stages of neurulation. We observed a number of defects in both wild type and Bent tail embryos, including a kinked neural tube, tight amnion, delay in axial rotation and even malrotation. The severity or frequency of most defects, the delay in axial rotation excluded, was significantly higher in Bent tail embryos compared to wild type embryos. Other abnormalities were seen in Bent tail embryos only. These defects were related to anterior and posterior neural tube closure and resulted in exencephaly and a closure delay of the posterior neuropore, respectively. The exencephalic phenotype was further analyzed by light microscopy in ED14 embryos, showing disorganization and overgrowth in the mesencephalon and rhombencephalon. In conclusion, the anterior and posterior neural tube closure defects in the Bent tail are strictly linked to the genetic defect in this mouse. Other phenotypic features described in this study also occur in the wild type genetic background of the Bent tail strain. Apparently, the genetic background contains elements conducive to these developmental abnormalities. PMID- 14523649 TI - Flavonoid-related regulation of auxin accumulation in Agrobacterium tumefaciens induced plant tumors. AB - Agrobacterium tumefaciens-induced plant tumors accumulate considerable concentrations of free auxin. To determine possible mechanisms by which high auxin concentrations are maintained, we examined the pattern of auxin and flavonoid distribution in plant tumors. Tumors were induced in transformants of Trifolium repens (L.), containing the beta-glucuronidase ( GUS)-fused auxin responsive promoter ( GH3) or chalcone synthase ( CHS2) genes, and in transformants of Arabidopsis thaliana (L.) Heynh., containing the GUS-fused synthetic auxin response element DR5. Expression of GH3::GUS and DR5::GUS was strong in proliferating metabolically active tumors, thus suggesting high free auxin concentrations. Immunolocalization of total auxin with indole-3-acetic acid antibodies was consistent with GH3::GUS expression indicating the highest auxin concentration in the tumor periphery. By in situ staining with diphenylboric acid 2-aminoethyl ester, by thin-layer chromatography, reverse-phase high-performance liquid chromatography, and two-photon laser-scanning microscopy spectrometry, tumor-specific flavones, isoflavones and pterocarpans were detected, namely 7,4' dihydroxyflavone (DHF), formononetin, and medicarpin. DHF was the dominant flavone in high free-auxin-accumulating stipules of Arabidopsis leaf primordia. Flavonoids were localized at the sites of strongest auxin-inducible CHS2::GUS expression in the tumor that was differentially modulated by auxin in the vascular tissue. CHS mRNA expression changes corresponded to the previously analyzed auxin concentration profile in tumors and roots of tumorized Ricinus plants. Application of DHF to stems, apically pretreated with alpha naphthaleneacetic acid, inhibited GH3::GUS expression in a fashion similar to 1-N naphthyl-phthalamic acid. Tumor, root and shoot growth was poor in inoculated tt4(85) flavonoid-deficient CHS mutants of Arabidopsis. It is concluded that CHS dependent flavonoid aglycones are possibly endogenous regulators of the basipetal auxin flux, thereby leading to free-auxin accumulation in A. tumefaciens-induced tumors. This, in turn, triggers vigorous proliferation and vascularization of the tumor tissues and suppresses their further differentiation. PMID- 14523651 TI - Informed consent needed from students. PMID- 14523647 TI - The role of bisphosphonates in diseases of childhood. AB - Bisphosphonates are synthetic analogues of pyrophosphate that inhibit bone resorption by their action on osteoclasts. In recent years, bisphosphonates have been used in children for treatment of a growing number of disorders associated primarily with generalized or localized osteoporosis, metabolic bone diseases, heterotopic calcification in soft tissues, and for resistant hypercalcemia. In the present review we discuss the pharmacological aspects of bisphosphonates and related bone pathophysiology, review the pediatric literature on the role of bisphosphonates in childhood diseases and our experience with these drugs. The theoretical concerns of possible adverse effects of these drugs on the growing skeleton have not materialized in the limited pediatric clinical experience. Bisphosphonates provide the pediatrician with an opportunity to treat mineral and bone disorders of childhood which until recently did not have satisfactory therapy, at the same time, being aware of the theoretical concerns on microdamage accumulation in bone, bone quality and teratogenic potential of these drugs. PMID- 14523652 TI - Getting the frame straight. PMID- 14523650 TI - cDNA cloning and expression of isoflavonoid-specific glucosyltransferase from Glycyrrhiza echinata cell-suspension cultures. AB - A cDNA encoding UDP-glucose: formononetin 7- O-glucosyltransferase, designated UGT73F1, was cloned from yeast extract-treated Glycyrrhiza echinata L. cell suspension cultures using probes from Scutellaria baicalensis UDP-glucose: flavonoid 7- O-glucosyltransferase. The open reading frame of the UGT73F1 cDNA encodes a 441-amino-acid protein with a predicted molecular mass of 48.7 kDa. The deduced amino acid sequence showed that the protein is related to the stress inducible glucosyltransferases. UGT73F1 mRNA was not detected in untreated G. echinata cultures but was transiently induced by treatment with yeast extract. Recombinant UGT73F1 was expressed as a histidine-tag fusion protein in Escherichia coli and purified to near homogeneity by nickel chelate chromatography. The purified recombinant enzyme was selective for isoflavonoid, formononetin and daidzein as substrates, while flavonoids and various tested non flavonoid compounds were poor substrates. PMID- 14523653 TI - Medical education reform in Iran. PMID- 14523654 TI - Preferences for physician attire. PMID- 14523655 TI - Formulating clinical questions during community preceptorships: a first step in utilizing evidence-based medicine. PMID- 14523657 TI - The punk. PMID- 14523656 TI - An interactive approach to teaching practice management to family practice residents. AB - Three years ago, our residency program began a new approach to teaching practice management to our second- and third-year residents. The underlying principles for the new curriculum involved a realization that our residents lacked basic business understanding and that they would likely learn more effectively through a hands-on approach. The new curriculum, which we describe in this article, is in large part built around the establishment of a mock practice during the second year of residency. Although the curriculum is still evolving, initial response and evaluation have been encouraging. PMID- 14523658 TI - Body image. PMID- 14523659 TI - Tuesdays with Morrie versus Stephen Hawking: living or dying with ALS. PMID- 14523660 TI - Tying it all together? A competency-based linkage model for family medicine. AB - Residency faculty in all specialties will be required by the Accreditation Council for Graduate Medical Education (ACGME) to fully implement competencies into residency programs by 2006. Understanding the new requirements is complicated by having several sets of guiding documents from different sources, including the general competencies of the ACGME, the Residency Review Committee for Family Practice requirements, the competencies developed by the Society of Teachers of Family Medicine, and the Recommended Curriculum Guidelines for Family Practice Residents by the American Academy of Family Physicians. A competency linkage model brings together the various guidelines and shows specifically how they are related. This model helps family practice residency faculty better understand the guiding expectations for their programs and develop more appropriate learning objectives and assessment methods. PMID- 14523661 TI - Unmet need for chronic disease and end-of-life care at urban family health centers in the Bronx, NY. AB - BACKGROUND AND OBJECTIVES: With population demographic shifts and as people live longer with progressive incurable illnesses, there will be a growing need for community-based chronic disease and end-of-life care services. Family practice sites could be strategic community-based settings in which to offer chronic disease and end-of-life care. This study determined whether two urban family health centers were meeting the chronic disease and end-of-life care needs of health center users, their families, and the local community. METHODS: Using a brief structured interview instrument, we interviewed a convenience sample of 218 patients at two hospital-affiliated family health centers in the Bronx, NY, about their recent experiences with serious illness and death of immediate family members. In addition, we developed geographically defined medical service areas for each health center and compared the age distribution of residents of the geographic areas with that of actual health center users. RESULTS: Of 218 health center users, 18% (n=39) had experienced the death of an immediate family member in the past year, of whom only 8% (n=3) stated that their family member had received end-of-life/chronic disease care at the health center, and only 23% (n=9) knew that their family member could have received such care. Of 26% (n=56) of study subjects who reported having an immediate family member who was currently seriously ill, 18% (n=10) stated that their family members were receiving care at the health center, and 23% (n=13) knew that their family members could have received such care. Eighty percent (n=174) of study subjects stated that they themselves would be interested in receiving end-of life/ chronic disease care at the health center in the event that they needed such care. The age distribution of health center users significantly differed from that of the residents of the geographically defined medical service areas, with an underrepresentation of people over the age of 75 among the health center users. CONCLUSIONS: We identified a potential unmet need for chronic disease and end-of life care among families of health center users and the larger community served by two family health centers in the Bronx, NY. Despite the frequent occurrence of serious illness and recent death among close family members, relatively small proportions of these family members had received care at the health centers, and few actual users were aware that such services could be provided there. Moreover, there was a significant demographic mismatch in age distribution between health center users and the surrounding community. These findings signal an important opportunity for the discipline of family medicine and its training programs to define and respond to the chronic disease and end-of-life care needs of patients and their families in community-based settings. PMID- 14523662 TI - Assessing students' communication and interpersonal skills across evaluation settings. AB - BACKGROUND AND OBJECTIVES: Medical students' interpersonal and communication skills are a fundamental dimension of their clinical competence and will be measured on the anticipated US Medical Licensure Examination (USMLE) standardized patient (SP) exam. We compared students' performance on measures of SP satisfaction on a third-year family medicine Objective Structured Clinical Examination (OSCE) with measures of SP satisfaction on a fourth-year Clinical Practice Examination (CPX). METHODS: A total of 127 students completed both the clerkship OSCE and a CPX. The CPX was a pilot of the National Board of Medical Examiners Standardized Patient Exam. To assess students' interpersonal skills, both exams used modified versions of the American Board of Internal Medicine Patient Satisfaction Questionnaire. Students' scores were standardized, and correlations were calculated. A logistic regression model examined the ability of the OSCE to predict poor performance on the CPX. RESULTS: The correlation between the OSCE and CPX patient satisfaction scores was.08. There was no significant predictive ability of the OSCE for poor performance on the CPX. CONCLUSIONS: Our study calls into question the ability of a routine end-of-clerkship OSCE to identify students' interpersonal skills abilities on fourth-year clinical performance exams and potentially that component of the anticipated USMLE SP exam. PMID- 14523663 TI - Refugee health and medical student training. AB - BACKGROUND AND OBJECTIVES: Cultural awareness training is an increasingly important priority within medical curricula. This article describes an academic family practice-community partnership focusing on health care needs of refugees that became the model for a medical school selective on cultural sensitivity training. METHODS: The monthly Refugee Health Night program featured dinner with preceptors and patients, international sessions on special medical needs of refugees, and actual clinical encounters with patients. Students were not expected to become culturally competent experts but, rather, health care providers sensitive to and appreciative of cultural context, experience, and expectations. We worked with students to develop sensitive methods of inquiry about mental health, especially around issues of war and torture. We used problem based cases to emphasize primary care continuity and the benefit of establishing trust over time. RESULTS: Over 2 years, 50 students and nearly 300 refugees (more than 73 families) participated. Students reported that their interactions with the refugees provided positive learning experiences, including expanded knowledge of diverse cultures and enhanced skills for overcoming communication barriers. Patients of refugee status were able to have emergent health care needs met in a timely fashion. CONCLUSIONS: Providing health care for refugee individuals and families presents many challenges as well as extraordinary opportunities for patients and practitioners to learn from one another. PMID- 14523664 TI - Racial and ethnic disparities in immunizations: recommendations for clinicians. AB - There continue to be significant racial and ethnic disparities in rates of childhood and adult immunizations in the United States. The causes are multifactorial, including inequities in education, income, and socioeconomic status; structural and systemic barriers in the health care delivery system; and beliefs, preferences, and practice patterns of the recipients and providers of care. Elimination of these disparities is a targeted priority in Healthy People 2010. The individual clinician can contribute to the narrowing of this gap by being informed of and using available national and regional resources, implementing national standards for culturally and linguistically appropriate health care services, and using every clinical encounter to assure that vaccination is offered and provided. Specific action steps are suggested. PMID- 14523665 TI - The family in the family medicine literature. AB - BACKGROUND: The degree to which the family medicine literature articulates unique family practice approaches and perspectives that are distinct from other primary care specialties has not been well studied. To determine how often a family practice perspective is included in clinical articles in the family medicine literature, we performed a family medicine journal review. METHODS: We reviewed 173 original research articles, review articles, and case reports from four family medicine journals from the year 2000: American Family Physician, Journal of Family Practice, Journal of the American Board of Family Practice, and Archives of Family Medicine. Each article was scored by independent reviewers as to whether the article discussed family aspects of care or family context. RESULTS: A total of 9.3% of clinical articles discussed family aspects of care; an additional 17.3% made some limited mention of the family or social context. Female first authors were 2.5 times more likely to have demonstrated a family practice perspective. Articles published in the American Family Physician and review articles were most likely to discuss family medicine perspectives, but these differences were not statistically significant. CONCLUSIONS: Unique family medicine approaches and perspectives are not discussed commonly in clinical articles in the family medicine literature. PMID- 14523666 TI - Implementation of an open access scheduling system in a residency training program. AB - BACKGROUND AND OBJECTIVES: Open access is one method of improving the quality of clinical practice. Leaving the majority of appointments open to be scheduled the same day allows patients to have control of their access to care. These appointments can be used for all visit types, including physical exams. Our objective was to implement this system to improve efficiency, and patient/provider satisfaction, while maintaining financial profitability. METHODS: We implemented open access at our academic practice on January 2, 2002. Clinical teams are essential for continuity, and implementation required extensive patient, provider, and staff education. We prebook the first three patient appointments for patients requiring language interpretation, prearranged transportation, and procedures. A phone triage system is still necessary for clinical questions or hesitancy about the system. RESULTS: Charges and revenues have increased since implementing the new system, as have patient satisfaction, visits per full-time equivalent, and total monthly volumes. Patient satisfaction is improved, as measured by the percent of abandoned phone calls, average time to make appointments, and the number of no-shows. Nursing work, as measured by triage, nursing callbacks, and bumped appointments, has declined markedly. CONCLUSIONS: Open access has improved revenue, simplified office processes, decreased nursing work, and improved patient satisfaction without any increase in provider time or clinic expansion. PMID- 14523667 TI - GP to FP to GP? PMID- 14523674 TI - Mechanisms of hepatocyte injury, multiorgan failure, and prognostic criteria in acute liver failure. AB - Acute liver failure (ALF) occurs when the rate and extent of liver cell death are not adequately balanced by regenerative activity. Two forms of liver cell death are recognized: apoptosis and necrosis. A number of causes of ALF have been shown, predominantly in experimental animal models, to induce one or the other form of liver cell death. Nonetheless, an insult capable of inducing apoptosis may cause cell death by necrosis, particularly if the degree of mitochondrial damage is sufficient to exhaust stores of adenosine triphosphate. Here we consider mechanisms of liver cell injury in ALF, including evolving knowledge of signaling pathways leading to hepatocellular apoptosis and necrosis. Factors that have an impact on the adequacy of hepatic regeneration along with the pathophysiology of complicating multiorgan failure are also reviewed. Prognostic criteria are discussed, especially in relation to current concepts of mechanisms of liver cell death and multiorgan dysfunction. PMID- 14523675 TI - Acute liver failure in the United States. AB - In the last 5 years the use of a multicenter approach has helped to define acute liver failure (ALF) in the United States. Drug-related hepatotoxicity comprises more than 50% of cases of ALF, including acetaminophen toxicity (40%) and idiosyncratic drugs (approximately 12%). Nearly 20% of cases remain of unknown etiology. Outcome of ALF is determined by etiology; by the degree of hepatic encephalopathy present on admission; and by complications, principally infection. More than 43% survive without a transplant, 28% die, and 29% undergo liver transplantation. Liver support machines have had no impact on this condition to date. A trial of N-acetylcysteine for the treatment of ALF not related to acetaminophen toxicity is underway. Future research in ALF in the United States should focus on limiting the number of cases related to drugs, searching for causes of the indeterminate cases, and developing more effective temporary liver support. PMID- 14523676 TI - Changing patterns of causation and the use of transplantation in the United kingdom. AB - Acute liver failure (ALF) is a rare condition in the United Kingdom. Comprehensive supportive intensive care of extra-hepatic organ failure and the early recognition of and use of transplantation for those who will not survive form the cornerstone of its management. Over the last 30 years there has been a reduction in the proportion of cases resulting from viral and seronegative hepatitis, and a progressive rise in those resulting from severe acetaminophen induced hepatotoxicity. The latter cases mostly result from deliberate self poisoning and formed the major cause of ALF hospital admissions and indication for emergency liver transplantation. The increasing misuse of acetaminophen has paralleled a rise in sales and greater availability of the drug. Introduction of legislation to restrict sales of acetaminophen has been followed by a fall in hospital admissions resulting from self-poisoning, a 20% reduction in deaths, and a 50% fall in the number of patients undergoing emergency liver transplantation. The reduction in acetaminophen-related ALF has been paralleled by an increase in the number of transplants performed in ALF of nonacetaminophen etiologies. PMID- 14523677 TI - Current issues and treatment of fulminant hepatic failure including transplantation in Hong Kong and the Far East. AB - In the Far East, fulminant hepatic failure is mainly due to viral hepatitis. In areas where hepatitis B infection is endemic, exacerbation of chronic hepatitis B infection, either spontaneously or on withdrawal of immunosuppressive therapy, is the major cause of fulminant hepatic failure. For hepatitis B surface antigen (HBsAg)-positive patients treated with intense immunosuppressive or cytotoxic therapy, preemptive use of lamivudine has drastically reduced the incidence of hepatitis due to hepatitis B exacerbation. Recently, the application of orthotopic liver transplantation, in particular living donor liver transplantation, has markedly improved the survival of patients with fulminant hepatic failure. In Hong Kong, the phenomenon of adoptive transfer of immunity to hepatitis B virus in liver transplantation has recently been reported. The mechanisms by which transfer of immunity occurs and its potential relationship with grafts from living related donors should be further explored. PMID- 14523678 TI - Molecular neurobiology of acute liver failure. AB - Acute liver failure results in encephalopathy and brain edema that is characterized by astrocytic cell swelling. Molecular biological techniques have led to the identification of alterations in expression of several genes coding for key astrocytic proteins in acute liver failure. Such proteins include amino acid transporters, structural proteins, the endothelial cell glucose transporter GLUT-1, the mitochondrial "peripheral-type" benzodiazepine receptor, and the water channel protein aquaporin IV. Magnetic resonance spectroscopic studies reveal increased brain lactate concentrations that are positively correlated with severity of encephalopathy and brain edema in acute liver failure, suggesting a deficit of cellular oxidative capacity and impending brain energy failure. Mild hypothermia prevents brain edema in acute liver failure, and mechanisms responsible for this beneficial effect include reduced blood-brain ammonia transfer as well as normalization of astrocytic amino acid transport and brain energy metabolism. Further elucidation of the molecular mechanisms responsible for brain edema and encephalopathy in acute liver failure will undoubtedly lead to novel treatment strategies for these complications. PMID- 14523679 TI - Pathogenesis of hepatic encephalopathy in acute liver failure. AB - Hepatic encephalopathy (HE) in acute liver injury signifies a serious prognosis. Brain edema and intracranial hypertension are major causes of death in this syndrome. Comparison of HE in acute liver failure (ALF) with that of cirrhosis allows recognition of important differences and similarities. A key role for ammonia in the pathogenesis of both HE and brain edema is now firmly supported by clinical and experimental data. Additional factors, such as infection, products of the necrotic liver, and synergistic toxins, may contribute to an altered mental state. A low plasma osmolarity, high temperature, and both high and low arterial pressure may affect brain water content. A combined derangement of cellular osmolarity coupled with cerebral hyperemia can explain the development of brain edema in ALF. Increasingly, study of the mechanisms responsible for brain swelling provides critical information for understanding the pathogenesis of HE. PMID- 14523680 TI - Intracranial hypertension in acute liver failure: pathophysiological basis of rational management. AB - Increased intracranial pressure (ICP) in patients with acute liver failure (ALF) remains a major cause of morbidity and mortality. Conventional methods of ammonia reduction such as the use of lactulose do not improve outcome, and metabolic substrates such as L-ornithine L aspartate may offer more promise. Mannitol remains the mainstay of therapy. An important role for cerebral hyperemia in the pathogenesis of increased ICP has led to a reevaluation of established therapies such as hyperventilation, N-acetylcysteine, thiopentone sodium, and propofol. Recent studies have focused on the role of systemic inflammatory response in the pathogenesis of increased ICP and support the use of antibiotics prophylactically. Moderate hypothermia reduces ICP in patients with uncontrolled intracranial hypertension and prevents increases in ICP during orthotopic liver transplantation (OLT). Advances in understanding the pathophysiological basis of intracranial hypertension in ALF have outstripped appropriate testing of the newly generated ideas in appropriate clinical trials, and more effort should be mounted at a national level to organize the appropriate multicenter studies required. PMID- 14523681 TI - New liver support devices in acute liver failure: a critical evaluation. AB - Acute liver failure remains a condition with substantial mortality in spite of the best available medical care. Liver transplantation is often not possible or delayed, and liver support systems can be used to maintain the patient during the crisis by bridging to transplantation or allowing recovery of the native liver. Bioartificial devices, using porcine or human hepatocytes, have yielded encouraging results on consciousness levels in a number of studies and have generally been found to be safe, although in one large controlled trial reported to date, benefit was restricted to certain subgroups. Artificial devices, like those based on albumin dialysis, are undoubtedly effective in removing protein bound toxins, and uncontrolled evidence shows some survival benefit. The large multicenter controlled trials necessary for proving the value of these systems are difficult to design. They have to take into account the influence of transplantation, etiology, and other factors on outcome. Achieving a standard medical regime for these critically ill patients between different centers is also difficult. PMID- 14523682 TI - Histopathological features of L-asparaginase-induced liver disease. AB - We studied the histopathological changes of liver in four patients who developed hepatomegaly and abnormal liver chemistry tests 2 to 20 days following administration of L-asparaginase as a part of a combination chemotherapy regimen for treatment of acute lymphoblastic leukemia. The severity of the liver disease due to L-asparaginase was unpredictable. One patient developed acute fulminant hepatic failure and required liver transplantation. The most consistent pathological change, observed in all four cases, was diffuse steatosis. Other changes included patchy hepatocyte necrosis, mixed inflammatory cell infiltrates in the portal tracts, and variable degrees of hepatocellular, or canalicular cholestasis, or a combination of these. PMID- 14523684 TI - [Effects of contemporaneous control of DRG-relevant coding by physicians]. AB - OBJECTIVE: In a prospective study we analysed the quality of ICD-coding in clinical everyday life of a department for internal medicine. METHODS: A skilled intern--the so-called DRG assistant--was temporarily released from clinical work. Over nine weeks he had to control all diagnoses contemporaneously that were ICD coded by his colleagues on admission and discharge of their patients. The DRG assistant had to ask for missing or correct implausible diagnoses, or inappropriate ICD-coding and with it also train his colleagues in appropriate coding. The effects of the DRG-assistant's correction of coding, on DRG consistent grouping and on the potential financial loss or benefit generated by his work were recorded. After stoppage of this control in a subsequent phase of the study the effect of the absence of the DRG-assistant, the absence of reminders and coding control and the changes of the clinic's revenue were determined. RESULTS: Corrections of ICD-coding by the DRG-assistant alone caused a remarkable increase in case-mix-index (CMI). CMI's mean value increased from 1.76 to 1.84 and the clinic's revenue increased by 180 Euro per patient (a total of about 80,000 Euro in nine weeks). After the end of the control, the case-mix index dropped within three weeks down to 1.14, corresponding with a potential loss of 1200 Euro per patient (assuming that patients' morbidity was the same over the time of the study). Coding corrections could not improve CMI in this situation. CONCLUSION: Contemporaneous control of ICD-coding by physicians seems to be essential in DRG based accounting. PMID- 14523685 TI - [American cutaneous leishmaniasis: special features in diagnosis and therapy]. AB - HISTORY AND CLINICAL FINDINGS: Three weeks after returning from a trip to Guatemala, a 33-year-old man developed two ulcers with indurated edges on his right leg and painful lymph nodes in the right groin. His general condition was not impaired. EXAMINATIONS: Histological examination revealed cellular infiltrates of the corium by lymphocytes and plasma cells, always accompanied by epithelial cells and multinuclear giant cells. Special stainings were unable to detect pathogens but Leishmania brasiliensis was identified using PCR. The Leishmania culture remained negative. THERAPY AND COURSE: After 7-day intravenous therapy with 20 mg/kg/d pentostam (pentavalent antimonial compound), the patient developed gastrointestinal complaints, coupled with a marked elevation of transaminases. Therapy was discontinued until the transaminase values normalized, then continued in reduced dosage (12 mg/kg body weight) for 23 days. The ulcers and lymphadenitis healed under this therapy. CONCLUSION: The diagnosis of American cutaneous Leishmaniasis may be complicated by the relative lack of pathogens in the lesions. PCR diagnosis are very helpful here. The therapy must be systemic owing to the danger of progression to mucocutaneous Leishmaniasis. The standard therapeutic pentostam has, however, a high rate of side effects and administration is exclusively intravenous. PMID- 14523686 TI - [Radiological procedures in gastrointestinal bleeding--case report]. PMID- 14523687 TI - [Radiological procedures in gastrointestinal bleeding--diagnostics]. PMID- 14523688 TI - [Radiological procedures in gastrointestinal bleeding--treatment]. PMID- 14523692 TI - [Once more: the care duty of the physician in ambulatory administration of sedatives. Decision of the Federal Court, 4/8/2003-VI ZR 265/02]. PMID- 14523691 TI - [Smoking and body weight--a main topic]. PMID- 14523693 TI - [Can a patient do heavy physical work after myocardial infarction?]. PMID- 14523694 TI - [For how long has methotrexate therapy to be interrupted before vaccination with live vaccine?]. PMID- 14523695 TI - Seven-year trends in stroke rehabilitation: patient characteristics, medical complications, and functional outcomes. AB - Studies of stroke trends have focused primarily on incidence, mortality, and hospitalization rates. There has been little evaluation of changes over time in the common patient characteristics, medical comorbidities, and functional outcomes of patients. The present study evaluated changes during a 7-year period. We found that while demographic variables, stroke severity, and most stroke characteristics remained relatively stable, disability levels at admission and discharge decreased and frequencies of both medical tube usage and many secondary medical complications increased over time. These changes have important implications for the clinical management of stroke patients in rehabilitation and for the organization and financing of stroke rehabilitation programs. PMID- 14523696 TI - Brief cognitive assessment and prediction of functional outcome in stroke. AB - To evaluate the ability to predict outcome with a brief measure of cognitive ability, we tested consecutive admissions who received inpatient rehabilitation for stroke with the Repeatable Battery for Assessment of Neuropsychological Symptoms (RBANS). Six months later, 34 discharged patients were contacted by telephone and were interviewed using a battery of functional outcome and quality of life measures. Multiple regression analysis showed that inpatient RBANS indexes predicted cognitive disability 6 months later. The present findings support the use of cognitive evaluations of patients with acute stroke to assist with prediction of outcome to be used in treatment planning. PMID- 14523697 TI - Two approaches to treating unilateral neglect after right hemisphere stroke: a preliminary investigation. AB - PURPOSE: This preliminary study assessed the efficacy of two treatment methods for persistent unilateral or hemispatial neglect following right hemisphere stroke. METHOD: Two patients received an intervention focusing on the underlying impairment of attention during visual scanning. Two other participants received an intervention that provided repetitive practice during a functional task of oral reading. Standardized tests of attention and neglect were administered before and after treatment. Performance on a functional task assessing generalization of treatment was probed at every session. RESULTS: Results indicate no clear conclusions regarding the efficacy of either approach. Factors contributing to these equivocal findings are addressed. PMID- 14523698 TI - An educational intervention for stroke rehabilitation patients and their families: healthy living after stroke. AB - The education of stroke patients and their families on prevention of recurrent stroke is a rehabilitation challenge. The principal objective of this article is to share a process for creating a stroke education program that improves patient and caregiver knowledge of secondary prevention, stroke signs and symptoms, modifiable risk factors, and maintenance of functional independence. During stroke recovery, patient education is a continuous journey. The education program needs to take into consideration the individual's readiness to learn, unique learning styles, and timing of sessions. In our educational program, Healthy Living After Stroke, we used the stroke team to develop different venues of learning. PMID- 14523700 TI - Physiological effects of exercise on stroke survivors. AB - PURPOSE: This study investigated the physiological responses of stroke survivors to a 16-week program of moderate aerobic and anaerobic exercise. METHOD: Forty participants who were a minimum of 6 months post stroke underwent physiological testing over the course of 2 days that included a maximal VO(2) stress test, a Biodex strength assessment, and a blood lipid panel. The participants were then randomly assigned to two groups, an aerobic training only (ATO) group and an aerobic and strength training (A&ST) group. Both groups were then required to exercise aerobically for 20 minutes for 3 days a week at a moderate intensity for 16 weeks. Only the A&ST group was required to do a series of eight strength training activities as part of their exercise protocol. Once the 16-week protocol was completed, all tests were repeated. These results were compared to the pretest data. RESULTS: Statistically significant changes in VO(2max) were noted in the A&ST group, while positive but not significant changes were noted in the ATO group. Both groups saw significant increases in knee flexion but not in knee extension. The A&ST group showed significant increases in both shoulder flexion and extension, while the ATO group only had significant increases in shoulder extension. CONCLUSION: We concluded that aerobic exercise produced positive metabolic changes in both groups, but only the A&ST group had statistically significant changes. Both groups demonstrated significant changes in functional strength, but the A&ST group experienced larger increases, likely due to the strength component of their training protocol. PMID- 14523699 TI - Meeting the needs of the community: a project to prevent stroke in Hispanics. AB - The Hispanic population is the fastest growing minority population in the United States. Little research has been done to address the stroke risk in the Hispanic population that has a higher prevalence of diabetes, alcohol use, and physical inactivity. To change behavior and decrease the risk of stroke, a comprehensive program that combines education, risk identification, and follow-up is needed. The purpose of this article is to identify the increased risk factors of stroke in the Hispanic population, identify the barriers to health care, and outline the development of a stroke education and screening program for Hispanics. PMID- 14523701 TI - Telephone group intervention for older stroke caregivers. AB - PURPOSE: This study evaluated the effectiveness of telephone groups for older, spousal caregivers of stroke survivors. METHOD: The 88 caregivers were mostly white females who were 70 years old on average and who had been providing care for an average of 3 years. Participants were randomized to treatment or control conditions, followed for 6 months, and assessed for depression, burden, loneliness, stress, and competence. Treatment participants engaged in an eight session psychoeducational telephone group. RESULTS: Treatment participants showed decreased stress over time but were not significantly different from control participants in the amount of change in stress. Control participants showed a significant increase in burden during the study; treatment participants showed a significant increase in competence. PMID- 14523702 TI - Reconceptualizing poststroke activity level using the Frenchay Activities Index. AB - Modifications of the Frenchay Activities Index were implemented in a study of postacute stroke rehabilitation outcomes. These modifications incorporated the prestroke activity levels and postrehabilitation goals and assessments of activity importance of 136 persons with stroke who received services in either a single modality or comprehensive outpatient rehabilitation setting. Prestroke activity levels were assessed retrospectively at admission to postacute rehabilitation, and current activity levels were assessed at discharge and 3 months post discharge. Prestroke activity levels were used to determine postrehabilitation goal attainment. Results showed that activities for which goals were less likely to be attained were more strenuous in nature, and activities for which goals were more likely to be attained were more sedentary in nature. The importance persons placed on specific activities was used to tailor the estimates of overall activity levels to reflect their importance to each person. Results showed that adjusting for importance decreased the estimate of prestroke activity level but had little effect on poststroke activity level. Incorporation of these modifications represents a reconceptualization of poststroke activity levels that clinicians can use to guide their selection of postacute rehabilitation therapy goals. PMID- 14523703 TI - The challenges of human subject research in the new millenium. PMID- 14523704 TI - The birth of the National Aphasia Association's Response Center. PMID- 14523705 TI - The domain management model: organizing care for stroke survivors and other persons with chronic diseases. AB - The US health care system is poorly designed to help patients with stroke and chronic health conditions and their functional consequences. These patients face multiple clinical problems. The Domain Management Model (DMM), based on the biopsychosocial model, describes four domains into which all patient and family problems can be classified. These problems are then managed over time through attention to care processes and outcomes. The use of the DMM in health care may help relieve patient and family suffering, guide resource use more efficiently, and improve satisfaction of health care providers (clinical, administrative, and financial) in treating complex patients with chronic health conditions. PMID- 14523706 TI - Communication among levels of care for stroke patients. AB - Stroke patients in the United States frequently progress through several different locations of care and groups of providers during the course of their recovery. Consistently effective communication from one level of care and one set of providers to the next is a basic expectation, but one that is often not fully met. New concepts and promising technologies offer opportunities to improve hand off processes at all levels. PMID- 14523707 TI - Clinical information systems for primary care: more than just an electronic medical record. AB - Massachusetts General Hospital Senior Health has integrated a computer-based clinical information system into all aspects of its patient care. This technology includes a longitudinal medical record but has broader functionality that encourages communication, education, and quality improvement activities. Care of geriatric patients is often challenging because of the complex nature of their clinical presentations. The case presented here, a patient at risk for stroke, illustrates how a clinical information system can facilitate quality care. The benefits, as well as difficulties, experienced as the system was integrated into the program's clinical practice are also described. PMID- 14523708 TI - The domain management model: a tool to help organize care of stroke survivors in skilled nursing facilities. AB - More rehabilitative care takes place in nursing homes than in any other setting. Skilled nursing facilities (SNFs) face increasing challenges in delivering stroke rehabilitation care. The Domain Management Model (DMM) organizes care of stroke survivors by identifying four critical domains that include all problems patients and families may face. Problems identified through the Minimum Data Set and Resident Assessment Protocols can be organized by the domains as well. In the SNF setting, the DMM can help organize rehabilitation care by assisting in the identification of all relevant clinical problems and by serving as a structured format for team meetings, problem management, and discharge summaries. PMID- 14523709 TI - Bridging to family and community support for older adults and the domain management model. AB - Older adults with chronic illness and functional impairment require continuous and long-term monitoring and supervision. Changes in the health care system make this harder to accomplish. Social work assistance can help identify issues and provide help through counseling, patient advocacy, resource utilization, and education. Tools that help include the Domain Management Model and well-organized team care. This article describes these interventions in a senior primary care practice and community-based screening and education program. Both programs bridge from acute illness care to improved chronic illness management in the community. PMID- 14523710 TI - Integration of a patient's care within a health care network. AB - Integrated networks add value to patient care through several mechanisms--sharing information and practice consistency based on evidence and access to a wide range of physicians and patient care venues. Successful networks are tied together through shared information systems and electronic medical records. The Domain Management Model provides a systematic structure for thinking about patients, especially those with physical disabilities and chronic illnesses, and can help improve care organization and documentation within networks. PMID- 14523711 TI - Ethical issues in rehabilitation: a qualitative analysis of dilemmas identified by occupational therapists. AB - Little available research exists to address the range of ethical issues encountered by occupational therapists (OTs) in their daily clinical work. The few articles in the literature have tended to be case-based or anecdotal or have focused on professional issues rather than clinical issues. To characterize the array of clinical ethical issues in occupational therapy, we asked OTs in a free standing academic rehabilitation hospital to describe in some detail up to three scenarios or situations from their clinical practice that raised morally troubling questions. A coding system was developed to preserve the richness of the detail but to allow for some categorization of the topics. A second section of the survey asked the therapists to rate whether various traditional ethics topics were of high, medium, or low interest to them. A third section asked that they identify the formats that would be most appealing to them for future educational interventions. Of the 56 therapists surveyed, 38 (or 68%) responded. The three self-generated topics mentioned most frequently by the therapists were (in decreasing order of frequency): reimbursement pressures, conflicts around goal setting, and patient/family refusal of team recommendations. The respondents were particularly interested in knowing more about patient-centered ethics topics, such as conflict resolution between teams and patients and the patient's role in decision making. Furthermore, they indicated a strong preference for interdisciplinary and interactive educational formats. PMID- 14523712 TI - Building a web of support for caregivers of persons with stroke. AB - Stroke is the leading cause of disability in the United States, with a cost of 30 billion US dollars annually. Most persons with stroke return home after initial treatment and are cared for by family members. Few researchers, however, have focused on Web-based interventions to strengthen caregivers'efforts. This article describes the development of Caring~Web, an online, in-home intervention of support for caregivers of persons with stroke. Through specifically designed Web pages, caregivers are able to ask questions of a nurse specialist and rehabilitation team, discuss issues with other caregivers and the nurse via email, and obtain educational information on stroke. PMID- 14523713 TI - Modern stroke unit. AB - The modern stroke unit is making significant contributions to the care of stroke victims and is proving to be an effective, cost-saving enterprise. The precise factors that contribute to the efficacy of these units have yet to be identified, but a combination of protocolized approaches to patient care, critical paths, a focus of expertise, and heightened index of suspicion for comorbidities all probably play a role. This article outlines the basic features of a modern stroke unit and surveys the literature on stroke unit outcomes. PMID- 14523714 TI - Influence of specific stroke diagnosis on prognostication, recovery, and early management. AB - Strokes are focal lesions in the brain and as such bring about deficits and portend a course of recovery depending on the nature of the lesion. Given the large number of possible combinations of locations, sizes, and types of lesions, a clear and reliable formula for predicting outcomes based on these characteristics has been statistically difficult, particularly early in the course of recovery. Severity of deficit and age of the patient accounts for sufficient variability that the influence of other factors becomes statistically harder to illustrate. PMID- 14523715 TI - Swallowing, nutrition, and hydration during acute stroke care. AB - Dysphagia occurs in up to half of patients after an acute stroke and may cause dehydration, undernutrition, and aspiration pneumonia. Current evidence suggests that a systematic program of diagnosis and treatment of dysphagia in an acute stroke management plan may yield dramatic reductions in aspiration pneumonia rates. There is also some evidence that nutritional supplementation and proper hydration may reduce morbidity and mortality in acute stroke patients. This article focuses on the recent advances in the evaluation and management of dysphagia, undernutrition, and dehydration related to acute stroke. A summary of pertinent studies in the area of stroke dysphagia and nutrition is also included. PMID- 14523716 TI - Mental status and behavioral changes in the early postacute phase of care. AB - Neurocognitive sequelae of stroke create a significant impact on the patient and frequently disrupt recovery from the physical manifestations. This article strives to review types of neurocognitive changes after stroke and discuss the underlying pathophysiology. Management options are presented for the diverse changes that may be encountered. PMID- 14523717 TI - Rehabilitation interventions during acute care of stroke patients. AB - The immediate care of a stroke patient admitted to hospital is best provided in a dedicated stroke unit, within which all of the key components of care can be coordinated. Neurologic diagnosis and intervention and general medical care are essential elements of acute stroke management. However, optimal outcome requires a comprehensive and multidisciplinary approach, which includes rehabilitation interventions. During the initial phases of care, rehabilitation interventions are mostly passive and emphasize prevention of secondary co-impairments such as contractures, pressure ulcers, and deconditioning. Rehabilitation interventions should be incorporated into care protocols for all patients and should begin immediately. As the patient becomes stable, more intensive therapy can be initiated in preparation for transition into the postacute phase of active rehabilitation. PMID- 14523718 TI - Ethical conflicts in the vocational rehabilitation of stroke survivors. PMID- 14523719 TI - Psychological stress and coping: framework for poststroke psychosocial care. AB - The ability to deal with and manage a chronic condition such as stroke requires a learning process embedded in personal experience that is often emotionally taxing as well as physically fatiguing. Stroke survivors typically need help in balancing demands and resources and in preventing or effectively coping with stress emotions to conserve energy. The major constructs outlined in this article provide a framework for psychosocial care to promote the experience of wellness in stroke survivors. PMID- 14523720 TI - Analysis of problem types and difficulty among older stroke caregivers. AB - Stroke onset causes disruption of family roles as caregiving tasks are assumed. The current study presents a qualitative and quantitative analysis of the problems of 123 older, spousal stroke caregivers. Problems were coded into 14 categories; frequencies of occurrence and difficulty were also analyzed. Differences in problem difficulties were examined by race, gender, years caregiving, and care receiver's functional level, depression, burden, and loneliness. Noncompliance was the least frequent but most difficult problem type. Lack of social involvement was the most frequent problem type and the most difficult for the more depressed, burdened, and lonely caregivers. This increase in problem specificity and detail of analysis has provided direction for clinical application in counseling the spouses of stroke survivors on how to manage the stress of their new life role. PMID- 14523721 TI - Predictors of emotional distress, general health, and caregiving outcomes in family caregivers of stroke survivors. AB - Predictors of emotional distress, general health, and stroke-related caregiving outcomes were determined in 104 family caregivers of stroke survivors based on a conceptual model derived from Lazarus' theory of stress and coping. Predictors of emotional distress (R(2) =.48, p <.001) were low caregiver self-esteem, high task difficulty, and high threat appraisal. Predictors of poorer health (R(2) =.25, p <.001) were not living with the patient, low household income, and high threat appraisal. Predictors of poor stroke-related care-giving outcomes (R(2) =.45, p <.001) were emotional distress, low benefit appraisal, high task difficulty, and high threat appraisal. Findings suggest potential areas for multidimensional caregiver interventions. PMID- 14523722 TI - Adaptation to stroke: a longitudinal study of depressive symptoms, physical health, and coping process. AB - The purposes of this study were to describe the natural history of adaptation to stroke and to identify survivor and caregiver predictors of depressive symptoms. Data were collected for 53 stroke survivors at four times from acute rehabilitation (T1) to 2 years post discharge (T4). Significant improvement occurred in depressive symptoms and perception of health, but change in the rate of depression was not significant. Family functioning became less healthy. Significant predictors of greater levels of depressive symptoms were lower levels of total support, finding meaning, and avoidance coping (T1) and lower levels of family functioning and belonging support (T4). Treatments to promote adaptation can be developed based on these predictors. PMID- 14523723 TI - Perceived needs of stroke survivors from non-English-speaking backgrounds and their family carers. AB - The purpose of this article is to discuss current findings in the research literature on the experiences and needs of stroke survivors and their family carers and to provide suggestions for future research. Based on this critical review, knowledge gaps and issues in stroke management in the community indicate that the needs of people surviving a stroke, particularly people from non-English speaking backgrounds, are not being adequately met by community-based health services. There is a critical need for changes in practices to meet the needs of this specific population. PMID- 14523725 TI - Counting my blessings as a stroke survivor. PMID- 14523724 TI - Coping with stroke: a family's perspective. PMID- 14523726 TI - Computer-assisted motivating rehabilitation (CAMR) for institutional, home, and educational late stroke programs. AB - Based on our results during the last 25 years, we are developing late stroke computer-assisted motivating rehabilitation (CAMR) for the upper extremity. Evidence has been accumulating that functional gains are possible even many years after the damage. However, postacute rehabilitation must be motivating and related to real-life functional activities, or it may fail to enlist active participation. With CAMR programs, such as briefly reported here, instead of exercise, the patient is engaged in a game (e.g., ping-pong); instead of concentrating on the specific movements, he/she is concentrating on the game and the movements become subconscious. Patients, even those who initially consider that they cannot accomplish the task, show interest and improvement, and functional recovery appears to be extended beyond the specific movements that are being practiced. CAMR is also suitable for late functional reorganization programs in an educational model. PMID- 14523727 TI - Virtual reality applications for motor rehabilitation after stroke. AB - Hemiparesis is the primary physical impairment underlying functional disability after stroke. A goal of rehabilitation is to enhance motor skill acquisition, which is a direct result of practice. However, frequency and duration of practice are limited in rehabilitation. Virtual reality (VR) is a computer technology that simulates real-life learning while providing augmented feedback and increased frequency, duration, and intensity of practiced tasks. The rate and extent of relearning of motor tasks could affect the duration, effectiveness, and cost of patient care. The purpose of this article is to review the use of VR training for motor rehabilitation after stroke. PMID- 14523728 TI - Neuromuscular electrical stimulation for motor restoration in hemiparesis. AB - This article assesses the clinical efficacy of established neuromuscular electrical stimulation (NMES) technologies for motor restoration in hemiparesis and provides an overview of evolving technologies. Transcutaneous NMES facilitates motor recovery. However, its impact on physical disability remains uncertain. Transcutaneous NMES also decreases shoulder subluxation, but its effect on shoulder pain remains uncertain. Clinically deployable upper extremity neuroprosthesis systems will not be available until sometime in the distant future. However, there is stronger evidence for the clinical utility of lower extremity neuroprosthesis systems. Evolving technology utilizes semi-implanted or fully implanted systems with more sophisticated control paradigms. Initial experiences with these systems are reviewed and directions for future research are discussed in this article. PMID- 14523729 TI - Robotic devices for movement therapy after stroke: current status and challenges to clinical acceptance. AB - Robotic devices for movement therapy are moving closer to becoming commercially available tools for aiding in stroke rehabilitation. Robotic technology offers a range of functions that will augment current clinical practice by leveraging therapists' time, cost effectively extending therapy programs, providing new measures of impairment, and offering new therapy protocols. In this article, we review work from several research laboratories that supports the clinical value of stroke therapy systems. A commercialization effort based on these results is described. We also discuss challenges to achieving clinical acceptance and practical implementation of these devices. PMID- 14523731 TI - Telerehabilitation tools for the provision of remote speech-language treatment. AB - Telerehabilitation is the method of using communication technologies to provide rehabilitation at a distance. Advancements in videoconferencing and networking technologies present opportunities to deliver rehabilitation services to patients at home, at school, in the workplace, and in the community. Speech-language treatment is an area of rehabilitation well suited for telerehabilitation. This article will review the technology for performing remote speech-language diagnosis and treatment for patients after stroke. Development of a telerehabilitation system incorporating computerized therapy tasks with a touchscreen interface is described. Such a system enhances off-the-shelf videoconferencing equipment and extends the potential utility and effectiveness of telerehabilitation sessions. PMID- 14523730 TI - Robot-aided neurorehabilitation: from evidence-based to science-based rehabilitation. AB - There is no "magic bullet" in rehabilitation. In the absence of direct neural transplants, neurological rehabilitation is an arduous process. We have pioneered the clinical application of robotics in stroke rehabilitation and have shown evidence of the positive impact of targeted exercise on stroke recovery. In this article, we will review results obtained in the initial clinical trials with 96 stroke patients at the Burke Rehabilitation Hospital. We will provide evidence that robot-aided training enhances recovery, that this enhanced recovery is sustained in the long term, and that this recovery is not due to a general physiological improvement--in fact, it appears to be limb and muscle group specific. An evidence-based approach must now segue into a more scientific approach to stroke rehabilitation. Given the length of the required protocols and patients' variability and limited census, the practical limitations of the evidence-based approach are self-evident and extend trials for years. Each patient and lesion is unique in stroke rehabilitation, so there is no reason to believe that a "one-size-fits-all" optimal treatment exists. To optimize therapy for individual patients, we need science-based models. In this article, we will summarize the scientific tools and models that we are investigating and present some of the results to date. PMID- 14523732 TI - Tracking gaze of patients with visuospatial neglect. AB - Video eye-tracking appears to have great utility in the investigation and assessment of visuospatial neglect, and it may also have important benefits to intervention. Applications include the use of desktop eye-tracking to quantify neglect and its sensitivity to social cues, acquisition of eye-tracking data during performance of clinical and experimental assessment tasks, and trials of a wireless system to track the gaze of patients carrying out activities of daily living. Displays that provide differential feedback contingent on gaze locus may be particularly applicable to computer-aided training. If eye-tracking proves to be useful for monitoring and for automatic delivery of training, it can eventually be incorporated into telerehabilitation. PMID- 14523733 TI - Waiting for a miracle. PMID- 14523734 TI - Theoretical and practical considerations in the restoration of function after stroke. AB - Recovery of function after stroke appears to include elements of both restoration and compensation. The brain is highly plastic, which allows reorganization after damage. Stroke produces permanent damage to the brain, so recovery must be based on activity in surviving cells that are either adjacent, contralateral, or in a different region. Furthermore, representation of a particular function in the brain is usually not limited to a single brain region. Multiple representation provides the opportunity for brain reorganization; functions are assumed by surviving brain structures. Compensation can be the initial response in the recovery phase and may persist through later phases because of the new habits formed (as the restraint therapy studies of Taub and others would suggest), because the damage to the brain is extensive and hinders restoration, because of secondary pathology, such as tendon shortening and muscle wasting, that does not allow brain reorganization to be translated into functional recovery, or because of inadequate (especially late) rehabilitation. PMID- 14523735 TI - Constraint-induced therapy approach to restoring function after neurological injury. AB - Traditional rehabilitation programs for persons with neurologic dysfunction emphasize a compensation, true recovery, or substitution approach to improve functional abilities. Constraint-induced (CI) movement therapy substantially increases more-affected extremity use in the life situation due to the influence of two different underlying mechanisms: overcoming learned nonuse and inducing use-dependent cortical reorganization. In this way, it bypasses the debate over whether a compensation, true recovery, or substitution approach should be used. The purposes of this article are to describe the CI therapy approach, discuss its proposed modes of action, and discuss other unique aspects of CI therapy as a rehabilitation technique. PMID- 14523736 TI - Activity-dependent factors affecting poststroke functional outcomes. AB - Over the last several years, there has been increasing recognition of the potential for central nervous system (CNS) recovery after brain damage. One commonality across the recovery and brain plasticity literature is that practice induces plastic, dynamic changes in the CNS. However, more than simply repetition, it is the manipulation of specific practice variables that appears to drive these dynamic processes in the CNS. The experimental manipulations used in the studies on neuroplasticity largely derive from the concept that in the undamaged or healthy brain neuronal connections and cortical maps are continuously remodeled by experience and by the performance of specific, intensive, and complex movements used to solve motor problems and attain goals. Intervention designed to promote recovery rather than compensation after stroke would then manipulate these same practice variables that have consistently promoted behavioral recovery and neuroplasticity in laboratory settings. Three current intervention strategies that incorporate these practice variables are reviewed. Preliminary results provide evidence that manipulation of task intensity and specificity and the sensorimotor experience of the task training are the necessary ingredients for maximizing the tremendous potential for recovery in patients with stroke. PMID- 14523737 TI - Effect of neurofeedback on motor recovery of a patient with brain injury: a case study and its implications for stroke rehabilitation. AB - This case study showed the effect of neurofeedback (NFB) training in a patient with a brain tumor and co-existing traumatic brain injury. The patient received 40 sessions of NFB intervention. Tests and videotaped recordings evaluated pre- and post-NFB intervention. This study demonstrated minimal to significant improvements in several functional tasks. The conclusion is that the use of NFB for a person with a head injury and brain tumor can be generalized to be used with stroke survivors. PMID- 14523738 TI - Mental practice: a promising restorative technique in stroke rehabilitation. AB - Upper limb hemiparesis is a common, yet debilitating, result of stroke. It has long been known that mental practice, when combined with physical practice, improves motor learning and performance. Recent studies also indicate that massed use of the affected arm results in cortical reorganizations and correlative functional improvements. During mental practice, there are widespread activations of neural and muscular mechanisms as if the arm were actually being used. This article introduces mental practice as a form of massed practice, reviews the bases for mental practice as a potent restorative technique, and presents data suggesting mental practice as a restorative technique for upper limb hemiparesis. PMID- 14523739 TI - Coherence in the urban family caregiver role with African American stroke survivors. AB - Expressions of coherence by urban caregivers in African American family systems are the focus of this ethnographic study. The profound changes that can accompany stroke may create considerable stress for families caring for the affected person. Coherence, a sense of mutuality, and differences in functioning among family members were discovered in this study to be important for maintaining stability within the family. Through coherence, family caregivers felt connected and this helped them survive in stressful times related to caring situations. The findings aid in defining coherence as an interpersonal process and place it in a family system context. PMID- 14523740 TI - An odyssey of hope: one man's journey through constraint-induced therapy. PMID- 14523741 TI - Recovery patterns of shoulder subluxation after stroke: a six-month follow-up study. AB - PURPOSE: Poststroke shoulder subluxation is a common complication that is thought to be irreversible without intervention. This study explores the natural recovery patterns of shoulder subluxation 6 months after stroke onset. METHOD: Ten first stroke survivors were evaluated in a free-standing rehabilitation hospital within 6 weeks of stroke onset and were reevaluated 6 months thereafter. Shoulder subluxation was measured in terms of glenohumeral asymmetries between affected and unaffected shoulders using anteroposterior radiographs before and after fitting of an appropriate support. Main outcome measures included shoulder subluxation asymmetries, range of motion, pain, and motor function as measured by the Fugl-Meyer Motor Function Assessment. RESULTS: Decreases in vertical asymmetry 6 months after initial evaluation were correlated with significant motor recovery (r =.7382, p =.018) and were associated with increases in the range of motion of shoulder abduction (r =.7167, p =.020). However, changes in vertical asymmetries were not correlated with changes in shoulder flexion (r =.3744, p =.286) or external rotation (r =.2155, p =.578) range of motion or with changes in shoulder pain (r = -.5189, p =.124). Six-month vertical asymmetries were correlated only with 6-month shoulder abduction range of motion (r =.6408, p =.046). CONCLUSION: Reductions in shoulder subluxation may occur spontaneously only when significant motor recovery of the affected upper limb occurs. PMID- 14523742 TI - The lighthouse strategy: Improving the functional status of patients with unilateral neglect after stroke and brain injury using a visual imagery intervention. AB - This study extends the work of J. Niemeier (1998) by using visual imagery for amelioration of the devastating impact of visual inattention or neglect on recovery from stroke or other brain injuries. Ten individuals with unilateral visual neglect who were undergoing acute physical rehabilitation after brain injury were cued by their interdisciplinary treatment team members to "be like" horizon-illuminating lighthouses and turn their heads left and right during functional and therapy training tasks. These treatment group patients, in comparison with waiting list controls, performed significantly better and more safely on route finding (p <.001), walking or wheelchair (p <.05), and problem solving (p <.05) tasks. The use of visual imagery techniques in rehabilitation of brain injuries is discussed. PMID- 14523743 TI - Conceptualization of a new stroke-specific outcome measure: the stroke impact scale. AB - Current stroke outcome measures are unable to detect some consequences of stroke that affect patients, families, and providers. The objective of this study was to ensure the content validity of a new stroke outcome measure. This was a qualitative study using individual interviews with patients and focus group interviews with patients, caregivers, and health care professionals. Participants included 30 individuals with mild and moderate stroke, 23 caregivers, and 9 stroke experts. Qualitative analysis of the individual and focus group interviews generated a list of potential items. Consensus panels reviewed the potential items, established domains for the measure, developed item scales, and decided on mechanisms for administration and scoring. Although the participants with stroke appeared highly recovered based on scores from conventional stroke assessments (Barthel Index and NIH Stroke Scale), stroke survivors and their caregivers identified numerous persisting impairments, disabilities, and handicaps. In general, stroke survivors described themselves as only about 50% recovered and reported that they had difficulty in activities in which they were not independent. To fully assess the impact of stroke on patients, we used the results of this qualitative study to develop a new stroke-specific outcome, the Stroke Impact Scale. PMID- 14523744 TI - A model of health environmental integration. AB - The spheres of health environmental integration (HEI) combine contemporary models of disease and disability and expand them with historic theories of the mind/body relationship. The basic premise is that neither the person nor his or her neurological sequelae can be characterized fully without viewing the environment as a dynamic system of which he or she and it are a part. Stroke rehabilitation based on such an integrative medical model could facilitate the health professional's understanding of how alternative treatments affect the patient's quality of life and experiences within environmental contexts. PMID- 14523745 TI - Recovery time of independent poststroke life habits: Part II. AB - The purposes of this study were to determine the time of the recovery of poststroke life habits and to identify prognostic indicators associated with recovery time among stroke patients in a rehabilitation program. A sample of 421 stroke patients who were admitted to a rehabilitation center was recruited from medical records available from January 1987 to December 1992. The relation between the achievement of independent life habits including bed mobility, transfers and ambulation, bathing activities, dressing activities, eating activities, home activities, sphincter control, and sleep with the potential covariates associated with recovery time was assessed through the analysis of survival data using the Cox maximum-likelihood proportional hazard models. The poststroke life habits obtained generated mean recovery times ranging from 5.51 to 57.60 days from admission to rehabilitation. The survival analysis revealed that the recovery time of the selected poststroke abilities was significantly influenced (p <.05) by one or several indicators; these included physical ability and neuropsychological and life habit characteristics. With this precious information, stroke rehabilitation specialists may be able to reduce the length of time required to recover independent poststroke life habits by treating the specific neuropsychological, physical, and life habit characteristics identified in this study. A faster poststroke recovery would reduce the socioeconomic impact generated by stroke disability and would also ensure a better quality of life to the stroke survivor. PMID- 14523746 TI - Artificial nutritional support: the daughters' dilemma. PMID- 14523747 TI - Upper-limb discoordination in hemiparetic stroke: implications for neurorehabilitation. AB - Clinically, upper-limb discoordination after stroke is evident in the form of stereotypic movement patterns that reflect a loss of independent joint control. These movement abnormalities, in conjunction with our recent quantitative findings under isometric conditions, provide evidence for an impaired capacity to generate certain muscle coactivation patterns in the impaired limb. In this article, we examine the parallels that exist between coordination disturbances observed under isometric and movement conditions. Our results suggest that discoordination in stroke may largely represent a manifestation of additional neural constraints on motor outflow. The neurotherapeutic implications of our findings are discussed. PMID- 14523748 TI - Mechanical measures of spasticity in stroke. AB - Mechanical measurements of stretch reflexes show promise for the quantification of spasticity after stroke. Mechanical measurements can increase the objectivity, repeatability, and precision of spasticity quantification and provide additional insight into the neuropathology that is not available with current clinical measurements. Several techniques are reviewed and the advantages and disadvantages of quantification of spasticity using mechanical measurements are considered. Although mechanical measures of spasticity offer many advantages, much research needs to be done to establish a standard measurement of spasticity that can be easily implemented in the clinic. PMID- 14523749 TI - Casting for the person with spasticity. AB - Casting is a fairly well-established option for the reduction of contractures due to spasticity, as well as a method for reducing tone. Some research suggests that casting is more effective than traditional therapy techniques in reducing contracture and is inconsistently better at decreasing hypertonicity. Traumatic brain injury and cerebral palsy are the most common diagnostic groups that are identified in the literature as benefiting from casting, but severe spasticity resulting from a stroke may also be positively affected by this technique. Several forms of casting are mentioned in the literature, including serial casting, inhibitive casting, drop-out casts, and bivalve casts. The purpose of this article is to review the differences and similarities between these kinds of casting and offer guidelines for their use in individuals who have suffered a stroke. PMID- 14523750 TI - Intrathecal baclofen therapy for stroke-related spasticity. AB - Intrathecal baclofen (ITB) therapy is a widely recognized management technique for severe, disabling spasticity in individuals with cerebral palsy and spinal and brain injuries. Its utility in the stroke population has only been recognized recently. Unlike the aforementioned patient populations, many stroke survivors are ambulatory and are able to maintain a certain degree of functional independence through compensatory use of the uninvolved limbs. Clinicians often fail to recognize the potential enhancement in the function of these individuals if they gain better control of their spastic limbs. Other spasticity treatments, such as oral medications and neurolytic procedures, offer the advantage of being nonsurgical; however, not every stroke patient will respond well to them. Some patients may not tolerate the systemic side effects of oral medications, such as drowsiness and sedation. In patients with severe multilimb spasticity, phenol and even high doses of botulinum toxin may not adequately control spasticity. ITB therapy offers the advantage of effectively decreasing severe, diffuse spasticity without causing untoward effects on arousal and cognition. This article will review the efficacy of ITB therapy in treating spasticity and enhancing function in stroke survivors. PMID- 14523752 TI - Withholding and withdrawing tube feeding for persons with dysphagia: exploring complications of advance directives and the obligations of a rehabilitation team. PMID- 14523751 TI - Effective use of chemodenervation and chemical neurolysis in the management of poststroke spasticity. AB - Chemodenervation with botulinum toxin (BTX) and chemical neurolysis with phenol can be used alone or in combination to effectively manage focal spasticity after a stroke. As with other treatments, these interventions must be done the right way, at the right time, in the right patient, and for the right reason. The proper use of these treatments requires careful patient assessment and realistic goals, knowledge of the peripheral functional anatomy, and an understanding of how these treatments work and how to best to administer them. This article reviews how BTX and phenol can be used in the management of poststroke spasticity. PMID- 14523753 TI - Recovery time of independent poststroke abilities: Part I. AB - The purposes of this study were to determine the time of the recovery of poststroke abilities and to identify prognostic indicators associated with recovery time among stroke patients undergoing a rehabilitation program. A sample of 421 stroke participants admitted to a rehabilitation center was recruited from medical records that were available from January 1987 to December 1992. The mean age was 61.8 years (range, 17-89 years). The relationship between the achievement of independent poststroke abilities and the potential covariates associated with recovery time was assessed through the analysis of survival data. Cox maximum likelihood proportional hazard models were used for the analysis. Independent poststroke abilities included behavior, cognitive, perceptual, communication, visual, and motor status. The time from rehabilitation admission to complete independence was introduced to the model in relation to the covariates. The mean time of recovery of poststroke abilities ranged from 18.70 to 32.40 days from the rehabilitation admission. The survival analysis revealed that the time of recovery of the selected poststroke abilities was significantly influenced (p <.05) by one or several factors, among these were neuropsychological, physical, and life habits. With this precious information, stroke rehabilitation specialists may be able to reduce the length of time required to recover independent poststroke abilities by treating the specific neuropsychological, physical, and life habit characteristics identified in this study. A faster poststroke recovery will reduce the socioeconomic impact generated by stroke disability and will ensure a better quality of life to the stroke survivor. PMID- 14523754 TI - The goal-planning process: a patient's perspective. PMID- 14523755 TI - Characteristics of gait in hemiplegia. AB - The following review examines the walking patterns of patients who have hemiplegia, primarily as a result of a stroke. Attention is given to the changes in the distance and temporal factors of walking, phasic patterns and joint ranges of motion throughout the walking cycle, the ground reaction forces, joint moments of force, joint powers, energy expenditure, and muscle activation patterns. The effect of orthotic intervention on these walking parameters is also addressed. A frequently cited issue regarding the gait patterns of these patients was that their walking patterns exhibit significant deviations from normal healthy individuals. Although hemiplegia is primarily associated with unilateral motor involvement, changes in almost all of the parameters used to assess walking were evident on both the involved and uninvolved sides of the body. Last, although hemiplegia appears to reflect a single diagnostic category, there is large interindividual variability in the patterns of gait deviations, which suggests that the management and treatment of these patients need to address the unique deficits of the individual. PMID- 14523756 TI - Biomechanics of ankle-foot orthoses: past, present, and future. AB - Pathological motion of the ankle-foot complex presents a major problem in the rehabilitation of stroke and head injury patients. For example, stroke patients often develop "drop foot," a problem involving excessive and uncontrolled plantar flexion. Ankle-foot orthoses (AFOs) are prescribed and used to restore normal motion or to constrain and inhibit abnormal motion. Research projects need to be conducted that include new methods of studies and experimental testing in which a better design could be achieved and appropriate prescription could be applied. PMID- 14523757 TI - Review of preorthotic biomechanical considerations. AB - The application of an ankle-foot orthosis in poststroke hemiplegia can improve the mechanics of ambulation and functional gait. However, prior to the fitting of an orthotic, the clinician should perform a thorough assessment of the biomechanical elements contributing to mobility problems within the patient. Many of these problems such as joint contracture or weakness should be addressed through physical therapy interventions prior to orthotic prescription. This article reviews the common biomechanical elements that contribute to mobility and gait disability in patients with stroke with special attention to orthotic management. PMID- 14523758 TI - Orthotic management in patients with stroke. AB - Orthotic management for the individual with a hemiplegic gait pattern secondary to a stroke is an important consideration for any treatment plan for this patient population. Patient evaluation, gait analysis, previous management, medical team cohesion, and patient expectations are elements necessary to establish the orthotic goals for the patient. These goals are the basis by which an orthotic treatment plan can be established as well as a measure for future outcome. Therefore, the purpose of this discussion is to describe the steps necessary to determine the most effective method of orthotic management and to identify orthotic design indications and contraindications. PMID- 14523759 TI - Addressing the needs of adolescent children when a parent becomes aphasic: one family's experiences. AB - There are several reports of the psychosocial effects of aphasia on family members and of factors that may influence a family's emotional adjustment. Most of these, however, have included only spouses or the adult children of individuals with aphasia. Therefore, to address how children of adolescent age or younger react to and cope with parental aphasia, this article provides an account of one family's experiences with stroke and aphasia. Specifically, the impact of a father's aphasia on his adolescent daughter as well as how the parents attempted to help their daughter adjust during acute and chronic recovery phases are described. PMID- 14523760 TI - A readability analysis of Australian stroke information. AB - The aims of this study were to document the readability of a stroke education brochure and to illustrate how the readability of this document might be improved. Readability analyses using three different formulas were conducted on a stroke education brochure from the Australian National Stroke Foundation. Consistent with previous studies of the readability of written health information, results indicated that the overall level of difficulty of the material was too complex for the average member of the public to read and understand. Revision of a randomly selected section of text using established guidelines for writing patient information, and subsequent reanalysis of this section, showed that simple revisions of health literature can dramatically improve readability. Given the importance of effective communication of preventative information about stroke, we present these results as a model of how future revisions of difficult to read stroke-related literature might be conducted to improve public education. PMID- 14523761 TI - Stroke: a bird's eye view. PMID- 14523762 TI - Guidelines for the selection of anti-infective agents for complicated intra abdominal infections. PMID- 14523764 TI - Long-term follow-up of women hospitalized for acute pyelonephritis. AB - Long-term outcome of acute pyelonephritis (AP) in adults is unknown. We evaluated the frequency of renal damage 10-20 years after hospitalization for AP in adult women and the utility of technetium Tc 99m-labeled dimercaptosuccinic acid (Tc 99m-DMSA) scanning for detection of renal scars; 63 of 203 women hospitalized with AP during 1982-1992 were included in the study. Tc 99m-DMSA scanning detected renal scarring in 29 women (46%). Multivariate analysis showed that pregnancy and hypoalbuminemia (albumin level, <3.2 g/dL) at hospitalization were independent risk factors for subsequent development of renal scars. At follow-up, hypertension was observed in approximately one-fifth of patients, regardless of renal scarring status. Four women with scars had a glomerular filtration rate of < or =75 mL/min; none of them developed severe renal impairment. In conclusion, the risk of developing renal scarring after AP in adult women is high. However, clinically relevant renal damage is rare 10-20 years after AP. Tc 99m-DMSA scanning is useful for detecting renal scars in adults but is not routinely needed in practice. PMID- 14523763 TI - Percutaneous injury, blood exposure, and adherence to standard precautions: are hospital-based health care providers still at risk? AB - To examine factors associated with blood exposure and percutaneous injury among health care workers, we assessed occupational risk factors, compliance with standard precautions, frequency of exposure, and reporting in a stratified random sample of 5123 physicians, nurses, and medical technologists working in Iowa community hospitals. Of these, 3223 (63%) participated. Mean rates of hand washing (32%-54%), avoiding needle recapping (29%-70%), and underreporting sharps injuries (22%-62%; overall, 32%) varied by occupation (P<.01). Logistic regression was used to estimate the adjusted odds of percutaneous injury (aOR(injury)), which increased 2%-3% for each sharp handled in a typical week. The overall aOR(injury) for never recapping needles was 0.74 (95% CI, 0.60-0.91). Any recent blood contact, a measure of consistent use of barrier precautions, had an overall aOR(injury) of 1.57 (95% CI, 1.32-1.86); among physicians, the aOR(injury) was 2.18 (95% CI, 1.34-3.54). Adherence to standard precautions was found to be suboptimal. Underreporting was found to be common. Percutaneous injury and mucocutaneous blood exposure are related to frequency of sharps handling and inversely related to routine standard-precaution compliance. New strategies for preventing exposures, training, and monitoring adherence are needed. PMID- 14523765 TI - Chloroquine versus sulfadoxine-pyrimethamine for treatment of Plasmodium falciparum malaria in Savannakhet Province, Lao People's Democratic Republic: an assessment of national antimalarial drug recommendations. AB - The in vivo efficacies of the Lao People's Democratic Republic (Laos) nationally recommended antimalarial agents--chloroquine and sulfadoxine-pyrimethamine-were assessed in a randomized, comparative trial that involved 100 patients with uncomplicated Plasmodium falciparum malaria who were followed for 42 days after starting treatment. Despite a shorter mean time to fever clearance associated with administration of chloroquine (mean time to clearance, 35.6 h; 95% confidence interval [CI], 26.3-45.0 h), compared with that associated with sulfadoxine-pyrimethamine (61.1 h; 95% CI, 50.9-71.3 h; P<.001), treatment failures were twice as frequent among patients receiving chloroquine therapy than among those receiving sulfadoxine-pyrimethamine therapy (36% vs. 18%; P=.02). Of 23 treatment failures, 10 (43%) were high grade. Treatment failure rates among children (age range, 5-15 years) were 4.9 times higher (95% CI, 2-12) than those among adults (P<.0001). Gametocytemia after antimalarial treatment was associated with receipt of sulfadoxine-pyrimethamine therapy and with treatment failure (P=.009). The efficacy of both chloroquine and sulfadoxine-pyrimethamine in Laos is unsatisfactory. PMID- 14523766 TI - Recurrent invasive pneumococcal disease: a population-based assessment. AB - We sought to define the risk of recurrence of invasive pneumococcal disease (IPD) and to define the characteristics of persons experiencing recurrent IPD through population-based surveillance. Cases of IPD were identified through the US Centers for Disease Control and Prevention's Active Bacterial Core Surveillance. Recurrent episodes were defined as isolation of Streptococcus pneumoniae from any normally sterile site > or =30 days after initial positive culture. Among 13,924 persons who survived their initial episode of IPD, 318 (2.3%) experienced > or =1 subsequent episode, for 376 total recurrences. The recurrence rate was 1294 episodes per 100,000 person-years, or 50 times the annual incidence of IPD. In multivariable analysis, a higher risk of recurrence was seen in persons infected with human immunodeficiency virus and in children <5 years old with chronic illness. Most (92%) persons with recurrence had a vaccine indication. The risk of recurrence among certain persons with IPD is extremely high. PMID- 14523767 TI - Two decades of disseminated tuberculosis at a university medical center: the expanding role of mycobacterial blood culture. AB - We describe the clinical presentation, predisposing conditions, diagnostic approach, and outcome for 52 patients with disseminated tuberculosis who presented at Duke University Medical Center (Durham, NC) from 1980 through 1999. The mean age of the patients was 52 years (range, 2-93 years). Fever and weight loss were common at presentation, and delays in the initiation of therapy often occurred. Predisposing conditions included human immunodeficiency virus infection (46% of patients), immunosuppressive therapy (21%), alcoholism (12%), diabetes mellitus (12%), and hematologic disorders (8%); 17% of patients had no disorder of immunity detected. Examination of biopsy specimens from sites of localized disease, especially lymph nodes, had a high diagnostic yield. In this study, mycobacterial blood culture appeared to be as sensitive as bone marrow culture in diagnosing disseminated tuberculosis (sensitivity, 58% vs. 54%). To diagnose disseminated tuberculosis, a search for sites of localized disease should be undertaken, and samples from these sites should be obtained. Mycobacterial blood culture can play an increasing role in the diagnosis of disseminated tuberculosis when localized disease is not found. PMID- 14523768 TI - West Nile encephalitis in 2 hematopoietic stem cell transplant recipients: case series and literature review. AB - Most human cases of West Nile virus infection are acquired via bites from an infected mosquito. In some cases, infection may also be transmitted by infected blood products or transplanted organs. There have been recent publications suggesting that chemotherapy and immunosuppression may increase a person's risks of developing central nervous system disease if the person is infected with the West Nile virus. Because patients undergoing hematopoietic stem cell transplantation not only are immunocompromised, but also receive multiple blood products, they are at a particularly high risk for acquiring symptomatic disease if exposed to the West Nile Virus. We describe here 2 patients who underwent hematopoietic transplantation at our institution and subsequently developed fatal West Nile virus infections. PMID- 14523769 TI - Severe Staphylococcus aureus infections caused by clonally related community acquired methicillin-susceptible and methicillin-resistant isolates. AB - We investigated the genetic relatedness of 5 community-acquired (CA) Staphylococcus aureus isolates obtained from 4 consecutive pediatric patients presenting with sepsis syndrome and severe pneumonia during a 3-week period in 2000. Two patients were infected with methicillin-susceptible S. aureus (MSSA), and 2 were infected with methicillin-resistant S. aureus (MRSA). The pulsed-field gel electrophoresis patterns for the 2 CA-MRSA isolates were identical to each other, as were the patterns for the 3 CA-MSSA isolates. A 2-band difference reflecting the presence of a staphylococcal cassette chromosome mec (SCCmec) element distinguished the CA-MRSA isolates from the CA-MSSA isolates. The small, mobile type IV SCCmec element was present in the CA-MRSA isolates. These data suggest that an insertion or, less likely, a deletion of the SCCmec type IV element occurred in a highly virulent S. aureus background. Staphylococcal toxin genes sea, seh, lukS-PV, and lukF-PV were detected in all isolates. Also, in all isolates, was a partial homolog of seo (seo'). The relationship among these patient isolates strengthens the assumption that CA-MRSA infections may be caused by isolates closely related to MSSA isolates. PMID- 14523770 TI - Randomized, double-blind, placebo-controlled trial to assess the rate of recurrence of oculorespiratory syndrome following influenza vaccination among persons previously affected. AB - During the 2000-2001 influenza immunization campaign in Canada, an oculorespiratory syndrome (ORS) was recognized as adverse event associated with one of the vaccines administered. The initial surveillance case definition for ORS in 2000-2001 specified onset within 24 h after vaccination and resolution within 48 h after onset; the restriction on the duration of ORS was removed from the case definition for the vaccine distributed during the 2001-2002 influenza season. The implicated vaccine contained large aggregates of unsplit virions; alterations to the manufacturing process for the vaccine distributed during 2001 2002 addressed this. A randomized, double-blind, placebo-controlled trial assessed the safety of the reformulated version in previously affected adults. The trial was halted early, after 61 participants had received an injection (34 had received vaccine, and 27 had received placebo). The vaccine-attributable recurrence rate for onset of ORS within 24 h after injection was 33% (95% confidence interval [CI], 10%-53%); for cases that resolved within 48 h, this rate was 27% (95% CI, 5%-47%). Previously affected persons should be informed of the risk of ORS recurrence but also that episodes of such recurrence are mild and well tolerated. PMID- 14523771 TI - Seroepidemiology of Helicobacter pylori and hepatitis A virus and the mode of transmission of infection: a 9-year cohort study in rural Japan. AB - We compared the seroepidemiologic patterns of Helicobacter pylori and hepatitis A virus (HAV) infections among participants in 2 independent cross-sectional studies conducted in Japan in 1986 and 1994. Subgroups were monitored with successive blood sampling. H. pylori and HAV infection status was defined by results of enzyme-linked immunosorbent assay. In 1986, the prevalence of H. pylori infection and HAV infection, respectively, were 80% and 70% among adults and 31% and 5% among children. The prevalence of both infections increased with age. Concordant infections were found in 74.5% of adults (kappa=0.2) versus 2% of children (kappa=0.05). During the 9-year study period, the incidence of H. pylori infection was 1.1% among adults and 2% among children. The seroprevalence of HAV remained constant. The disparity between the increase in prevalence of H. pylori and HAV infection with age is likely associated with improvements in hygienic practices. The discordance between the presence of the infections among younger persons is evidence against a common source and/or vehicle for transmission. PMID- 14523772 TI - Percutaneous aspiration-injection-reaspiration drainage plus albendazole or mebendazole for hepatic cystic echinococcosis: a meta-analysis. AB - Using meta-analysis methodology, we compared the clinical outcomes for 769 patients with hepatic cystic echinococcosis treated with percutaneous aspiration injection-reaspiration (PAIR) plus albendazole or mebendazole (group 1) with 952 era-matched historical control subjects undergoing surgical intervention (group 2). The rate of clinical and parasitologic cure (P<.0001) was greater in patients receiving PAIR plus chemotherapy. Disease recurrence (P<.0001), major complications (anaphylaxis, biliary fistula, cyst infection, liver/intra abdominal abscess, and sepsis; P<.0001), minor complications (P<.0001), and death (P<.0824) occurred more frequently among surgical control subjects. Fever (P<.002) and minor allergic reactions subjects (P<.0001) were more common among PAIR-treated subjects. The mean durations of hospital stay were 2.4 days for group 1 and 15.0 days for group 2 (P<.001). Compared with surgery, PAIR plus chemotherapy is associated with greater clinical and parasitologic efficacy; lower rates of morbidity, mortality, and disease recurrence; and shorter hospital stays. PMID- 14523773 TI - Rift Valley fever epidemic in Saudi Arabia: epidemiological, clinical, and laboratory characteristics. AB - This cohort descriptive study summarizes the epidemiological, clinical, and laboratory characteristics of the Rift Valley fever (RVF) epidemic that occurred in Saudi Arabia from 26 August 2000 through 22 September 2001. A total of 886 cases were reported. Of 834 reported cases for which laboratory results were available, 81.9% were laboratory confirmed, of which 51.1% were positive for only RVF immunoglobulin M, 35.7% were positive for only RVF antigen, and 13.2% were positive for both. The mean age (+/- standard deviation) was 46.9+/-19.4 years, and the ratio of male to female patients was 4:1. Clinical and laboratory features included fever (92.6% of patients), nausea (59.4%), vomiting (52.6%), abdominal pain (38.0%), diarrhea (22.1%), jaundice (18.1%), neurological manifestations (17.1%), hemorrhagic manifestations (7.1%), vision loss or scotomas (1.5%), elevated liver enzyme levels (98%), elevated lactate dehydrogenase level (60.2%), thrombocytopenia (38.4%), leukopenia (39.7%), renal impairment or failure (27.8%), elevated creatine kinase level (27.3%), and severe anemia (15.1%). The mortality rate was 13.9%. Bleeding, neurological manifestations, and jaundice were independently associated with a high mortality rate. Patients with leukopenia had significantly a lower mortality rate than did those with a normal or high leukocyte count (2.3% vs. 27.9%; odds ratio, 0.09; 95% confidence interval, 0.01-0.63). PMID- 14523774 TI - Transmission of influenza: implications for control in health care settings. AB - Annual influenza epidemics in the United States result in an average of >36,000 deaths and 114,000 hospitalizations. Influenza can spread rapidly to patients and health care personnel in health care settings after influenza is introduced by visitors, staff, or patients. Influenza outbreaks in health care facilities can have potentially devastating consequences, particularly for immunocompromised persons. Although vaccination of health care personnel and patients is the primary means to prevent and control outbreaks of influenza in health care settings, antiviral influenza medications and isolation precautions are important adjuncts. Although droplet transmission is thought to be the primary mode of influenza transmission, limited evidence is available to support the relative clinical importance of contact, droplet, and droplet nuclei (airborne) transmission of influenza. In this article, the results of studies on the modes of influenza transmission and their relevant isolation precautions are reviewed. PMID- 14523775 TI - Enfuvirtide (T-20): a novel human immunodeficiency virus type 1 fusion inhibitor. AB - The development of highly active antiretroviral therapy has improved life expectancy and reduced progression to acquired immunodeficiency syndrome in human immunodeficiency virus (HIV)-infected patients. However, resistance to currently available classes of antiretroviral drugs has become a problem, limiting the options for patients with advanced disease who have been heavily treated. Enfuvirtide (T-20; ENF), a synthetic peptide, is the first of a new class of antiretrovirals that block entry of virus into host cells. ENF interferes with conformational changes required for membrane fusion and injection of virus into the host cell. Optimal treatment of HIV infection will likely require combinations of drugs that target novel stages of HIV type 1 entry and replication. PMID- 14523776 TI - Acute meningoencephalitis in chronic human immunodeficiency virus (HIV) infection: putative central nervous system escape of HIV replication. AB - We describe 3 patients with chronic human immunodeficiency virus (HIV) infection who presented with syndromes compatible with acute meningoencephalitis secondary to HIV; these syndromes were characterized by elevated cerebrospinal spinal fluid (CSF) HIV viral loads and T2-weighted signal abnormalities on magnetic resonance imaging of the brain. After the initiation of or a change in highly active antiretroviral therapy (HAART), each of the patients had significant and rapid improvement in neurologic symptoms and dramatic reductions in CSF HIV viral loads. Although further investigation is needed, these findings suggest that measurement of CSF HIV viral load and treatment with central nervous system penetrating HAART should be considered for patients with acute neurologic complaints, chronic HIV infection, and no other identifiable cause of neurologic illness. PMID- 14523777 TI - Association between adherence to antiretroviral therapy and human immunodeficiency virus drug resistance. AB - Nonadherence to highly active antiretroviral therapy (HAART) is a major cause of human immunodeficiency virus (HIV) drug resistance; however the level of nonadherence associated with the greatest risk of resistance is unknown. Beginning in February 2000, 195 patients at the Johns Hopkins Outpatient Center (Baltimore, MD) who were receiving HAART and who had HIV loads of <500 copies/mL were recruited into a cohort study and observed for 1 year. At each visit, adherence to HAART was assessed and plasma samples were obtained and stored for resistance testing, if indicated. The overall incidence of viral rebound with clinically significant resistance was 14.5 cases per 100 person-years. By multivariate Cox proportional hazards regression, a cumulative adherence of 70% 89%, a CD4 cell nadir of <200 cells/microL, and the missing of a scheduled clinic visit in the past month were independently associated with an increased hazard of viral rebound with clinically significant resistance. Clinicians and patients must set high adherence goals to avoid the development of resistance. PMID- 14523778 TI - Long-term efficacy, safety, and tolerability of indinavir-based therapy in protease inhibitor-naive adults with advanced HIV infection. AB - A double-blind, randomized study of zidovudine-experienced, PI- and lamivudine naive adults with baseline CD4 cell counts of < or =50 cells/mm3 had demonstrated that the HIV suppression achieved with zidovudine, lamivudine, and indinavir therapy was superior to that achieved with dual-nucleoside or indinavir-only regimens after 24 weeks of therapy. In a 192-week extension of the study, 371 participants received open-label indinavir with or without other antiretroviral drugs. One hundred and eight subjects were originally randomized to receive triple therapy. After 216 weeks, the proportion of subjects with HIV RNA levels of <500 copies/mL were 34%, according to a general estimating equation analysis, 92%, according to an observed data analysis, and 24%, according to an intention to-treat analysis counting noncompleters as failures; the proportions of subjects with HIV RNA levels of <50 copies/mL were 31%, 85%, and 22%, respectively. Hyperbilirubinemia (experienced by 31% of subjects), nausea (17%), abdominal pain (14%), and nephrolithiasis (13%) were the most common drug-related adverse events during the extension. PMID- 14523779 TI - Blood cultures for women with uncomplicated acute pyelonephritis: are they necessary? AB - To assess the utility of blood cultures in the management of uncomplicated pyelonephritis in women, we prospectively collected data from 583 cases. Discordant cases were defined as those for which the pathogens isolated from urine and from blood were different. We found that 97.6% of cases were nondiscordant. Clinical and microbiological evolution of infection did not differ between the 2 groups, and no changes of antibiotic therapy were required on the basis of blood culture results. Blood culture may not be routinely required for the evaluation of uncomplicated pyelonephritis in women. PMID- 14523780 TI - Stenotrophomonas maltophilia intestinal colonization in hospitalized oncology patients with diarrhea. AB - A 6-week surveillance study was performed to determine the prevalence of Stenotrophomonas maltophilia intestinal colonization among patients hospitalized in an oncology unit who developed diarrhea. Ninety-two stool samples obtained from 41 patients were cultured, and 4 patients (4 [9.5%] of 41 patients) had cultures that were positive for S. maltophilia. After controlling for duration of diarrhea, patients colonized with S. maltophilia had received a greater number of different types of antibiotics than noncolonized patients (5 vs. 3 different drugs; P=.04). PMID- 14523781 TI - Ophthalmological and biological features of the oculorespiratory syndrome after influenza vaccination. AB - We report the ophthalmological and laboratory findings of 6 patients who, after influenza vaccination, were affected by oculorespiratory syndrome (ORS), complaining of red eyes, photophobia, blurred vision, palpebral edema, ocular pain and itching, and conjunctival secretions. The conjunctivae were mildly hyperemic with few follicles, but the ophthalmological examination findings were otherwise normal. Patients had lymphopenia and decreased levels of the total hemolytic complement and the third and fourth component of the complement. We conclude that ORS causes conjunctivitis and seems to involve the complement. PMID- 14523782 TI - Common adverse events associated with the use of ribavirin for severe acute respiratory syndrome in Canada. AB - Although information on efficacy and adverse drug reactions is lacking, ribavirin has been used empirically for the treatment of severe acute respiratory syndrome (SARS). We report common adverse events in 110 patients with suspected or probable SARS who were treated with ribavirin. Sixty-one percent of the patients had evidence of hemolytic anemia, and hypocalcemia and hypomagnesmia were reported in 58% and 46% of patients, respectively. PMID- 14523783 TI - Good interpretation of the results of a diagnostic test. PMID- 14523785 TI - Current spectrum of bacterial infections in patients with cancer. PMID- 14523786 TI - Nadir CD4+ T cell count predicts response to subcutaneous recombinant interleukin 2. AB - Community Program for Clinical Research on AIDS 059 was a multicenter study conducted among human immunodeficiency virus (HIV)-infected individuals with CD4+ cell counts > or =300 cells/mm3 who were randomly assigned to receive antiretroviral therapy with or without intermittent subcutaneously administered recombinant interleukin-2 (rIL-2). To identify factors associated with a response to IL-2, a secondary analysis was performed that included the subset of rIL-2 recipients who were able to complete all 3 initial treatment cycles. Predictors of a change in CD4+ cell count between baseline and 1 month after the start of treatment cycle 3 were examined in a multivariate model that included sex, race, body surface area, rIL-2 dose, HIV load, and both baseline and nadir CD4+ cell count. The combination of race and sex (P=.027) and the nadir CD4+ cell count (P=.005) were significant predictors of mean CD4+ cell count response. Baseline CD4+ cell count had no significant effect. The strong association between nadir CD4+ cell count and CD4+ cell count response suggests that immunologic losses resulting from HIV-mediated CD4+ cell depletion may be irreversible. PMID- 14523787 TI - Molecular confirmation of human alveolar echinococcosis in Poland. AB - Infections of humans with Echinococcus multilocularis, the causative agent of alveolar echinococcosis (AE), a zoonosis, have been described with increasing frequency in Poland since 1994. In the attempt to verify these reports, we analyzed specimens obtained from a representative group of Polish patients. Liver lesions in patients with AE that was diagnosed on the basis of results of histological and serological tests contained E. multilocularis DNA, as shown by the presence of specific microsatellite sequences and mitochondrial 12S rDNA. The same tests clearly distinguished between AE and cystic echinococcosis, which is caused by Echinococcus granulosus. These data are unequivocal proof that human infections with E. multilocularis occur in Poland. PMID- 14523789 TI - Oxaliplatin-safety profile: neurotoxicity. AB - Oxaliplatin has become an integral part of various chemotherapy protocols, and in advanced colorectal cancer in particular. While oxaliplatin has only mild hematologic and gastrointestinal side effects, its dose-limiting toxicity is a cumulative sensory neurotoxicity that resembles that of cisplatin with the important difference of a more rapid and complete reversibility. The reversibility of neurotoxicity has been assured in long-term follow-up of patients who have received adjuvant oxaliplatin-based chemotherapy. In addition, oxaliplatin causes a very unique, but frequent, acute sensory neuropathy that is triggered or aggravated by exposure to cold but is rapidly reversible, without persistent impairment of sensory function. Various strategies have been proposed to prevent or treat oxaliplatin-induced neurotoxicity. The "Stop-and-Go" concept uses the reversibility of neurologic symptoms to aim at delivering higher cumulative oxaliplatin doses as long as the therapy is still effective. Several neuromodulatory agents such as calcium-magnesium infusions, antiepileptic drugs like carbamazepine or gabapentin, amifostine, alpha-lipoic acid, and glutathione have shown promising activity in prophylaxis and treatment of oxaliplatin-induced neurotoxicity. However, larger confirmatory trials are still lacking so that, to date, no evidence-based recommendation can be given for the prophylaxis of oxaliplatin-induced neurotoxicity. The predictability of neurotoxicity associated with oxaliplatin-based therapy should allow patients and doctors to develop strategies to manage this side effect in view of the individual patient's clinical situation. PMID- 14523790 TI - Pharmacology of oxaliplatin in solid tumor patients with hepatic dysfunction: a preliminary report of the National Cancer Institute Organ Dysfunction Working Group. AB - Oxaliplatin, which was recently approved by the US Food and Drug Administration for the treatment of advanced colorectal cancer, is often administered to patients with altered liver function caused by hepatic metastases. Because of the absence of data on the clearance of oxaliplatin in patients with liver dysfunction, and to develop dosing recommendations for the safe use of oxaliplatin in this clinical setting, the Organ Dysfunction Working Group of the National Cancer Institute performed a phase I and pharmacokinetic trial of this drug in patients displaying a wide range of liver function abnormalities. Sixty patients grouped into five classes of liver function were enrolled in this study, including a 12-patient control group with normal liver function, 15 patients with "mild," 16 patients with "moderate," and 16 with "severe" liver dysfunction, as well as one patient who had undergone liver transplantation. The patients had a median age of 62 years and a Eastern Cooperative Oncology Group performance status of 0. All patients had received prior systemic chemotherapy, and 70% had a diagnosis of a gastrointestinal malignancy. The control patients were treated with 130 mg/m(2) oxaliplatin intravenously every 21 days; the starting doses for patients with mild, moderate, or severe liver dysfunction or post-liver transplantation were 105, 80, or 60 mg/m(2), respectively. Cohorts of three or more patients were escalated from these starting doses to 130 mg/m(2) if dose limiting toxicities were not observed, and pharmacokinetic evaluations were performed at each dose level for every category of liver dysfunction. We found that oxaliplatin was well tolerated at its recommended dose and schedule of 130 mg/m(2) every 21 days in patients with all levels of liver dysfunction, and that there was no apparent alteration in the clearance of either total or ultrafilterable platinum species from plasma, even in patients with severe hepatic functional abnormalities. PMID- 14523791 TI - Administration of oxaliplatin to patients with renal dysfunction: a preliminary report of the national cancer institute organ dysfunction working group. AB - Oxaliplatin is an approved agent with clinical activity in the treatment of advanced colorectal cancer. Preliminary pharmacokinetic evidence suggests that oxaliplatin is predominantly cleared by renal excretion; however, formal dosing guidelines in patients with renal impairment are lacking. The National Cancer Institute Organ Dysfunction Working Group initiated the following dose-escalation pharmacokinetic trial of oxaliplatin in patients with varying degrees of renal function. Thirty-seven patients with various solid tumor malignancies were stratified into four renal dysfunction groups based on their measured 24-hour urinary creatinine clearance (CrCL). Patients in group A (controls) all had a CrCL >or= 60 mL/min; group B patients had mild renal dysfunction with CrCLs ranging from 40 to 59 mL/min; group C patients had moderate renal dysfunction with CrCLs of 20 to 39 mL/min; and patients with a CrCL < 20 mL/min were entered into the group D severe cohort. The starting oxaliplatin dose for patients in the normal group A was 130 mg/m(2), while lower doses of 105, 80, and 60 mg/m(2) were used in groups B, C, and D, respectively. Patients received a 2-hour intravenous infusion of oxaliplatin every 3 weeks, and doses were escalated in cohorts of three patients in each renal dysfunction group in the absence of any severe dose limiting toxicity. Oxaliplatin-associated platinum pharmacokinetics were monitored in both plasma (bound + unbound) and plasma ultrafiltrates (unbound). Full single-agent doses of oxaliplatin of 130 mg/m(2) were well tolerated by patients with normal, mild, and moderate renal dysfunction in groups A, B, and C, respectively. Only one patient was enrolled in group D. Unbound platinum clearance significantly correlated with CrCL (r =.884), but the increased systemic exposures to circulating platinum were not associated with increased clinical toxicities. These data suggest that dose reductions of single-agent oxaliplatin are not necessary in patients with CrCLs >20 mL/min. PMID- 14523792 TI - Oxaliplatin-based combined-modality therapy for rectal cancer. AB - There are two conventional treatments for clinically resectable rectal cancer. The first is surgery, and, if the tumor is T3 and/or N1-2, this is followed by postoperative combined-modality therapy. The second, for patients with ultrasound T3 or clinical T4 disease, is preoperative combined-modality therapy followed by surgery and postoperative chemotherapy. In this review, the results of these approaches as well as novel combined-modality approaches using oxaliplatin-based regimens will be presented. PMID- 14523793 TI - Colorectal metastases confined to the liver: a unique opportunity? AB - Liver metastases nearly always represent disseminated cancer, and systemic therapies designed for controlling systemic tumors are usually indicated. However, in a minority of patients with colorectal cancer, management of the metastatic hepatic disease will be clinically important and even curative. This review discusses novel treatment approaches that may be indicated in colorectal cancer patients with liver metastases, including the potential use of systemic therapy as part of multimodality therapy in an effort to cure selected patients with colorectal liver metastases. PMID- 14523794 TI - Recent experience with oxaliplatin or irinotecan combined with 5-fluorouracil and leucovorin in the treatment of colorectal cancer. AB - During the last decade, considerable progress has been made in the development of 5-fluorouracil/leucovorin (5-FU/LV) regimens that optimize antitumor efficacy while minimizing toxicity in the management of colorectal cancer. The use of continuous infusions allowed the administration of high doses of 5-FU and LV, leading to enhanced efficacy with acceptable toxicity. Furthermore, the relatively low toxicity of such regimens as bimonthly 5-FU/LV has provided the opportunity to add other antitumor agents, oxaliplatin and irinotecan, with the possibility of improving tumor response rates and patient survival. The availability of these novel agents provides additional therapeutic options for tumors that are resistant to 5-FU/LV regimens. Several studies have shown that combinations with irinotecan or oxaliplatin have improved response rates and survival over 5-FU/LV regimens alone in first-line therapy of advanced colorectal cancer. Other trials currently in progress attempt to identify whether these new chemotherapeutic regimens can improve survival in the adjuvant setting. PMID- 14523795 TI - Pharmacogenomics in colorectal cancer. AB - The goal in administering chemotherapeutics is to develop the ability to predict the outcome of therapy in terms of response and toxicity. Technology has been developed to allow tumor profiling with the measurement of protein expression and gene expression levels of markers and even genetic polymorphisms that may predict response to particular chemotherapeutics. The chemotherapeutics for which particular markers have been shown to predict outcome include the fluoropyrimidines and platinums. The next step is to develop clinical trials that will prospectively assess the benefits of profiling a patient's particular tumor, which should translate into an improvement in response and toxicity. PMID- 14523796 TI - The experience with oxaliplatin in the treatment of upper gastrointestinal carcinomas. AB - The treatment for advanced gastroesophageal tumors is still largely based on 5 fluorouracil and cisplatin combinations, and the treatment for advanced pancreatic tumors principally relies on gemcitabine. The need for more active agents is evident in light of the poor outcomes for patients with these tumors. Oxaliplatin is an important chemotherapeutic agent currently being investigated in the treatment of gastrointestinal carcinomas. This diaminocyclohexane-platinum compound has significant differences from cisplatin and carboplatin with respect to its activity and toxicity. It appears to have activity in tumors believed to be marginally sensitive to the other platinum agents, and it is neither ototoxic nor nephrotoxic. The main side effect of oxaliplatin is a sensory neuropathy exacerbated by cold exposure. After extensive clinical development, oxaliplatin has become an important option in the treatment of colorectal cancer. Its role in the treatment of upper gastrointestinal cancers is less well established, but is the focus of intense investigation. Much of what we currently know is based on small phase I and II studies, but knowledge is rapidly accumulating. The fact that cisplatin remains an important drug in the treatment of several upper gastrointestinal cancers raises the hope that oxaliplatin will contribute significantly to the treatment of those malignancies as well. Larger randomized phase III trials are being designed and conducted to fully define the role of oxaliplatin in the treatment of upper gastrointestinal tumors in the relatively near future. PMID- 14523797 TI - Opportunities for newer agents in combination with oxaliplatin. AB - Oxaliplatin is a new third-generation platinum analogue with unique activity in colorectal cancer. This agent has changed the approach to chemotherapy for this disease. Novel biologic or molecular targeting agents are now under study in phase I to III clinical trials. Many of these agents are quite promising in colorectal cancer therapy. Most advanced among these include anti-vascular endothelial growth factor antibodies, antibodies directed against the epidermal growth factor receptor (HER-1), small-molecule tyrosine kinase inhibitors of epidermal growth factor receptor, and the proteasome inhibitor PS-341 (bortezomib). The rationale for combinations with oxaliplatin and early clinical trial results are discussed. PMID- 14523805 TI - Transplant immunology. AB - The immune response to an allogeneic transplanted organ is T-cell dependent. It is governed partially by the context in which the T-cell encounters the antigen and can range from apoptosis, anergy, and neglect to full activation. The current armamentarium of immunosuppressive agents acts to inhibit the various steps of this T-cell activation pathway; at the level of the T-cell receptor (monoclonal antibodies such as OKT3), intracellular signally (calcineurine-inhibitors), DNA synthesis (azathioprine), or to cause lymphocyte depletion (ATG, ALG). Most protocols use a combination of agents for induction and maintenance immunosuppression. Although successful in preventing and treating allograft rejection, they are not without side effects. With improved patient and graft survival rates, adverse events such as hypertension, nephrotoxicity, hyperglycaemia, and lymphoproliferative disease become increasingly important issues. Newer drugs (IL-2 receptor antagonists, mycophenolate mofetil, rapamycin) have been introduced in an attempt to spare or avoid these adverse effects. Inducing graft tolerance and long-term drug-free survival is the goal of transplant immunologists. Postulated mechanisms include clonal deletion, anergy, and immunoredirection. Although a number of methods have been tested experimentally, none has been proven to induce tolerance for routine clinical use. Immunosuppression remains the cornerstone of the success of organ transplantation. Until investigators are able to induce tolerance in their transplant recipients or develop a tolerance assay, they would need to continue to tailor their immunosuppressive therapy according to the risk profile of the individual recipient. PMID- 14523806 TI - The role of low-bandwidth telemedicine in surgical prescreening. AB - PURPOSE: Low-bandwidth, Internet-based telemedicine is an inexpensive technology that could help deliver heath care in medically underserved areas. The purpose of this study was to evaluate the usefulness of low-bandwidth telemedicine in remote surgical evaluation. METHODS: A group of surgeons and nurses traveled to Mombasa, Kenya to provide surgical assistance at the Coast Province General Hospital (CPGH). Before the visit of the surgical team, surgeons evaluated patients via low-bandwidth telemedicine. Prescreening was performed through use of an Internet based desktop computer system that was supported technically by the Virginia Commonwealth University (VCU) Medical Informatics and Technology Applications Consortium (MITAC) telemedicine laboratory. Surgical patients were evaluated remotely regarding appropriateness of surgical intervention by reviewing e-mails that included the patient's age, history, physical examination, digital images of the patient, and digital images of pertinent radiographs. RESULTS: Fifty-one patients, including 7 pediatric patients, were prescreened. Thirty-three of the 51 patients (65%) were deemed inappropriate for surgery before this trip because of advanced disease or absence of necessary local medical resources. Of the 18 patients determined to be appropriate candidates for surgery by remote prescreening, 18 (100%) were operated on successfully during the relief effort. Sixty patients including 9 (15%) pediatric patients underwent surgery over the course of 5 days in CPGH. Pediatric cases included various laparoscopic, oncologic, and soft tissue reconstruction. CONCLUSIONS: Low-bandwidth, Internet based telemedicine is a cost-effective technology that can efficiently and effectively prescreen surgical patients in remote areas. PMID- 14523807 TI - All-terrain vehicle rules and regulations: impact on pediatric mortality. AB - BACKGROUND/PURPOSE: All-terrain vehicles (ATV) use by children leads to severe injury and death. Since the US Consumer Product Safety Commission consent decree expired in 1998, there has been little movement in regulating ATV use for children (<16 yr). The authors hypothesized that states with laws and regulations restricting pediatric ATV use may abrogate excess death compared with states without such restrictions. METHODS: Pediatric mortality data reported to the consumer product safety commission from 1982 to 1998 were analyzed as well as state all-terrain vehicle requirements compiled by the Specialty Vehicle Institute of America in August 2001. The authors calculated ATV mortality rate by dividing ATV mortality frequency by 1980-2000 pediatric census results. They compared the top 26 states with the highest ATV mortality rates (TOP) with those of all other states (OTH) in terms of age, ATV type, ATV occupancy, and ATV laws. Chi-square analysis was performed. RESULTS: There were 1,342 ATV pediatric deaths during the 16-year period. The TOP states averaged approximately a 2-fold increase in adjusted ATV mortality rate compared with the national ATV pediatric mortality rate. Ninety-two percent of TOP states have no licensing laws compared with 73% of the OTH states (P <.07). There is no difference between groups with regard to minimum age requirements and safety certification. CONCLUSIONS: Current legal and regulatory standards have low probability of decreasing ATV-related pediatric mortality. States should adopt laws that restrict the use of ATV's for children less than 16 years of age and potentially prevent excess ATV-related pediatric mortality. PMID- 14523808 TI - The absence of lateral fusion in cloacal partition. AB - BACKGROUND/PURPOSE: The mechanism by which the cloaca becomes partitioned into a dorsal rectal part and a ventral genitourinary sinus has been the subject of speculation for more than a century. Despite repeated suggestions that partitioning of the cloaca by fusion of lateral folds does not occur, the concept continues to hold sway in many student and surgical texts. The authors reviewed the histologic and 3-dimensional appearance of the urorectal septum in human and rat embryos to see if there was any evidence of lateral fusion in its formation. METHODS: Sprague-Dawley rat embryos (n = 143) were examined between 11 and 21 days' gestation and compared with human embryo sections (57 embryos) held in historical collections in Europe. Rat embryos were examined by microscopy, dissection, and serial histologic section. In addition, some specimens were sectioned in wax until the lumen of the cloaca was reached, after which they were dewaxed and the internal cavities imaged with scanning electronmicroscopy (n = 18 of 143). RESULTS: Cloacal "partitioning" resulted from a combination of growth of the mesenchyme of the hindgut and genitourinary sinus, an alteration in the position of the cloaca in relation to surrounding structures secondary to growth in the ventral, infraumbilical abdominal wall and changes in the curvature of the developing spine, and apoptosis in the dorsal wall of the cloaca with shortening of the dorsal cloacal wall. There was no septum, as it is usually defined, between the developing bladder and hindgut. There was no evidence on either histologic section or scanning electronmicroscopy of any process of fusion occurring between the 2 lateral folds within the lumen of the cloaca. CONCLUSIONS: Lateral fusion of the side walls of the cloaca does not play a role in cloacal "partition." Development of the bladder and hindgut occurs by a process that involves growth, differentiation, and remodeling. PMID- 14523809 TI - Significance of heparin-binding growth factor expression on cells of solid pediatric tumors. AB - BACKGROUND: The heparin-binding growth factors pleiotrophin (PTN), midkine (MK), vascular endothelial growth factor (VEGF), and basic fibroblast growth factor (bFGF) stimulate tumor cell proliferation and angiogenesis. In this study the authors wanted to know if these growth factors are expressed by cell lines and tumor tissue of solid pediatric tumors, growth factor expression is influenced by proinflammatory cytokines, and local growth factor concentration has an influence on experimental tumor growth. METHODS: Growth factor mRNA expression was analyzed by reverse transcriptase polymerase chain reaction (RT-PCR) and protein secretion by enzyme-linked immunosorbent assay (ELISA). Neuroblastoma cells were suspended in solutions containing different growth factor concentrations before injection into the nude mice, which were given pentosan polysulfate (PPS) for antagonism. RESULTS: The analyzed growth factors were expressed by most cells of solid malignant pediatric tumors. Their expression was not influenced by proinflammatory cytokines. The inhibition of tumor growth by PPS in the nude mouse model was dependent on the local growth factor concentration. High concentration excluded significant tumor suppression. CONCLUSIONS: Because of the redundancy of growth factor expression and the abolishment of PPS efficacy by a high local growth factor concentration, the authors conclude that overall targeting of growth factors is a promising approach to cancer therapy in childhood. PMID- 14523810 TI - Tissue-engineered stomach: a preliminary report of a versatile in vivo model with therapeutic potential. AB - BACKGROUND/PURPOSE: Microgastria and postgastrectomy morbidities are substantial. The authors hypothesized a functional living tissue-engineered stomach could function as a replacement alternative. METHODS: Stomach organoid units, mesenchymal cores surrounded by epithelia, were isolated from neonatal and adult rats and transplanted paratopically on biodegradable polymer tubes, which were implanted in syngeneic hosts, varying the inclusion of stomach regions. Four weeks later, tissue-engineered stomach (TES) was either harvested or anastomosed. GFP labeling was performed before implantation. Histology and immunohistochemical detection of the antigensgastrin and actin smooth muscle were performed. RESULTS: Ninety-eight percent of all animals generated TES, including TES formation from adult tissue. Immunohistochemistry for alpha-actin smooth muscle and gastrin confirms the presence of a smooth muscle layer and a well-developed gastric epithelium containing all the elements of the native rat stomach including gastric pits and squamous layers, varying by included regions at harvest. TES architecture was maintained in anastomosis: GFP-labeled TES maintained signal in anastomosis, proving the donor origin of the TES. CONCLUSIONS: TES resembles native stomach and maintains robust histology in anastomosis, a new versatile model for the study of gastric physiology and possible therapy. PMID- 14523811 TI - Evidence that the notochord may be pivotal in the development of sacral and anorectal malformations. AB - BACKGROUND/PURPOSE: The notochord is known to organize normal development of central axial structures, such as the spinal cord, vertebral column, and anorectum, but its role in abnormal development of these organs has not been well documented. The current study has used Ethylenethiourea to induce anorectal malformations in fetal rats, allowing investigation of abnormalities of the notochord and their relationship to the axial structural abnormalities that occur. METHODS: Timed-mated pregnant rats were fed Ethylenethiourea by gavage on gestational day 10. Their embryos were harvested on gestational days 13 to 16 and sectioned in either the transverse or sagittal plane. Sections were stained with H and E and examined serially. RESULTS: Anorectal malformations were identified in 29 of 34 embryos and neural tube defects in 24, ranging from an accessory neural tube to lumbo-sacral rachischisis. There was no tail or only a rudimentary tail in the majority of embryos. Abnormalities of the notochord in the lumbo sacral area included ventro-dorsal branching, ventral deviation, and ectopic notochordal tissue. Most abnormal notochord branches and ectopic notochordal tissue were abnormally close to or in contact with the wall of the cloaca or neural tube. CONCLUSIONS: Given the known role of the notochord in controlling normal development, this study would suggest that abnormal notochord development may be pivotal in producing neural tube defects and anorectal malformations, possibly by altering sonic hedgehog signalling. PMID- 14523812 TI - Pediatric extraadrenal paraganglioma. AB - BACKGROUND: Pediatric paraganglioma is rare and extraadrenal paraganglioma has not been characterized in children. METHODS: The authors reviewed the medical records and pathology samples of children with extraadrenal paraganglioma treated at our institution between December 1978 and September 2000. Clinical presentation, treatment, and outcome were evaluated. RESULTS: Eight patients (median age, 11.4 years) were identified, 4 were boys and none had a family history of paraganglioma or associated syndromes. Primary tumors arose in the retroperitoneum (n = 3), carotid body (n = 2), jugulotympanic ganglion (n = 1), cervical-paraspinal region (n = 1), and lung (n = 1). Extraadrenal paraganglioma had not been suspected at presentation in any patient. Of 5 patients who underwent gross total resection at the time of diagnosis, 4 remain disease free, 1 had microscopic residual tumor and died of disease. Three patients had initially unresectable disease, 2 are disease free after neoadjuvant therapy and delayed surgery, and 1 has persistent disease after tumor embolization and radiotherapy. CONCLUSIONS: Pediatric extraadrenal paraganglioma occurs most commonly in the retroperitoneum and head and neck, and the diagnosis usually is not suspected at the time of presentation. Surgery is the mainstay of treatment, and outcome is good after gross total resection. Neoadjuvant therapy can facilitate complete resection of initially unresectable tumors. PMID- 14523813 TI - Analysis of the effects of endothelin-3 on the development of neural crest cells in the embryonic mouse gut. AB - BACKGROUND/PURPOSE: Mutations in the endothelin-3 (ET-3) and endothelin-B receptor (EDNR-B) genes cause terminal colonic aganglionosis in mice and are linked to Hirschsprung's disease. These experiments are designed to determine if the development of terminal enteric ganglia depends on changes in proliferation, apoptosis, or differentiation of enteric neural crest (NC) cells in response to ET-3. METHODS: Gut from embryonic lethal-spotted mice (lacking ET-3) and controls were investigated in vivo. NC-derived cells were identified immunohistochemically and their proliferation, apoptosis and differentiation monitored by bromodeoxyuridine incorporation, the terminal deoxytransferase poly dU nick end labelling (TUNEL) reaction, and appearance of neuronal nitric oxide synthase (NOS), respectively. RESULTS: No differences in apoptosis or proliferation of NC cells were apparent between lethal-spotted embryos and controls. Although no temporal differences in the differentiation of NOS neurones were evident, these cells appeared more cranially in the gut in the absence of ET-3 than in controls. CONCLUSIONS: ET-3 has no detectable influence on proliferation, apoptosis, or timing of differentiation of NC-derived cells in the gut. However, the more proximal location of differentiated neurones in the absence of ET-3 is consistent with a restricted role in migration of NC-derived cells. PMID- 14523814 TI - The management of old urethral injury in young girls: analysis of 44 cases. AB - BACKGROUND/PURPOSE: Traumatic urethral injury in girls is rare, and there is no consensus on its management. The authors report their 22-year experience. METHODS: Forty girls presented with urethrovaginal fistula. Twenty-six girls presented with cystostomy tube in place, whereas 17 girls presented with complete urinary incontinence. Incision and dilatation of the obliterated urethra was carried out in 7 patients. Vaginal repair of urethrovaginal fistula was performed in 4 patients. Transpubic reconstruction of the urethra using a modified Young Dees-Leadbetter procedure with simultaneous repair of the urethrovaginal fistula was performed in 35 patients (once in 27, twice in 5, and 3 times in 3 patients). RESULTS: Follow-up in 40 girls averaged 3.5 years. Twenty-nine patients have regained normal urinary control, and 11 patients have mild stress urinary incontinence. Four patients were lost to follow-up. CONCLUSIONS: Simple dilation of the obliterated urethra can reestablish satisfactory urethral patency if the obliterated segment is short. The vaginal approach to urethrovaginal fistula may be successful in patients without concomitant urethral stricture or in those with stricture amenable to simple dilation. The transpubic approach remains the method of choice for repairing complete urethral disruption and severe urethral stricture, especially when associated with urethrovaginal fistula. PMID- 14523815 TI - Treatment of infantile hypertrophic pyloric stenosis in a district general hospital: a review of 160 cases. AB - BACKGROUND: Ramstedt's pyloromyotomy has long been the standard operation for the treatment of infantile hypertrophic pyloric stenosis. Controversy exists over whether this procedure can be performed safely in the district general hospital setting or whether it should be restricted to specialist pediatric units only. METHODS: Retrospective analysis was performed on the medical records of a series of 160 infants treated by Ramstedt's pyloromyotomy by 2 surgeons in a district general hospital over 16 years. RESULTS: There was no perioperative mortality. Oral feeding was achieved by 24 hours in 76% of infants, and there was persistent vomiting in only 3.8%. Wound discharge was encountered in 4.4% and confirmed wound infection in 1.3%. Wound dehiscence occurred in 1.9% of infants. Inadvertent mucosal perforation occurred in 19% of cases, although all cases were recognized and repaired at once with no apparent ill effects. These results are comparable with those reported from specialist pediatric units and from pediatric surgeons working within general surgical units. CONCLUSIONS: Infantile hypertrophic pyloric stenosis can be treated safely in a district general hospital when care is provided by appropriately trained surgical, anesthetic, and pediatric staff. PMID- 14523816 TI - Transanal endorectal pull-through for Hirschsprung's disease: experience with 68 patients. AB - BACKGROUND/PURPOSE: The aim of this study was to evaluate the indications, results, and complications of transanal endorectal pull-through (TEPT) in the management of recto-sigmoid Hirschsprung's disease (HD). METHODS: Between November 1998 and March 2002, 68 TEPT procedures were performed in infants and children. The patients' ages ranged from 6 days to 13 years. The primary diagnosis in all 68 patients was Hirschsprung's disease confined to the recto sigmoid region. All children had their operations done without construction of preoperative colostomy except for one. Follow-up period ranged from 3 to 39 months (mean, 21 months). RESULTS: The mean operating time was 90 minutes, and average length of bowel resected was 25 cm. Sixty-two patients had satisfactory results without complications. Blood transfusion was needed in only 11 patients. Recovery was very fast, and patients often were hungry within 24 hours. Feeding was resumed within 48 hours. One patient required laparotomy during the procedure owing to injury to the urethra. Two patients required colostomy 3 and 5 days after surgery respectively, because of delayed leakage. Three patients suffered from attacks of enterocolitis 6 to 9 months postoperatively. There was increased frequency of defecation (5 to 15 times daily) for 4 to 6 weeks after surgery in all the patients. There was no constipation, no incontinence, no cuff abscess, and no mortality in any of the patients. Average frequency of defecation was 1 to 3 times daily after 3 months. The cost of the TEPT technique was almost half that of the open surgery. CONCLUSIONS: TEPT takes less time, has less bleeding, shorter hospital stay, less morbidity, and earlier recovery than similar open pull-through procedures. The hazards and morbidities associated with laparotomy and colostomy may be avoided with a one-stage technique in Hirschsprung's disease confined to the recto-sigmoid region. Careful long-term follow-up is required to assess continence and sexual function. PMID- 14523817 TI - Protective effect of polyunsaturated phosphatidylcholine on liver damage induced by biliary obstruction in rats. AB - BACKGROUND/PURPOSE: Persistent inflammatory response secondary to congenital or acquired biliary choleastasis plays an important role in the pathophysiology of hepatic tissue damage. The polyunsaturated fatty acids (PUFA) have been shown to suppress the inflammatory reactions in vivo and in vitro. PUFA has been shown also to protect against various types of experimental liver damage in animal models and isolated hepatocytes. Therefore, the aim of this study was to investigate the protective effect of PUFA administration on liver damage using the rat chronic biliary obstruction model. METHODS: Swiss albino rats of either sex were divided into 4 groups as follows: control group (group 1, 10 rats); rats with sham operation and treated with saline group 2, 10 rats); rats with biliary obstruction (group 3, 15 rats); and polyunsaturated phophatidylcholine (PPC) treated rats with biliary obstruction (Group 4, 15 rats). Biliary obstruction was induced by double ligation and division of the common bile duct. PUFA treatment was started 2 weeks later from biliary obstruction in doses of 50 mg/d per rat and continued for 2 weeks. All animals were killed after 4 weeks of common bile duct ligation or sham operation. Liver damage and cholestasis were determined by biochemical and histologic examinations. RESULTS: The data showed a decrease in plasma bilirubin level (both conjugated and unconjugated) and liver enzyme levels (AST, ALT, AP, GGT, 5'-NT) in group 4, when compared with group 3 (P <.05). Tissue levels of malondialdehyde (MDA) in group 4 was 20.00 +/- 2.93 compared with that in group 3, 27.12 +/- 2.96 (P <.05). Administration of PUFA to the biliary obstructed rats resulted in inhibition of collagen accumulation (P <.05) and ductal proliferation (P <.05). CONCLUSIONS: PUFA reduced liver damage, ductular proliferation, and fibrosis in biliary obstructed rats. These effects suggest that it might be a useful agent to preserve liver function in patients with biliary obstruction such as biliary atresia. PMID- 14523818 TI - Fetal tissue engineering: in vitro analysis of muscle constructs. AB - BACKGROUND/PURPOSE: This study was aimed at examining the impact of different tissue engineering techniques on fetal muscle construct architecture. METHODS: Myoblasts from ovine specimens of fetal skeletal muscle were expanded in culture and their growth rates determined. Cells were seeded at different densities onto 3 scaffold types, namely polyglycolic acid (PGA) treated with poly-l-lactic acid (PLLA), a composite of PGA with poly-4-hydroxybutyrate (P4HB), and a collagen hydrogel. Constructs were maintained in a bioreactor and submitted to histologic, scanning electron microscopy, and DNA analyses at different time-points. Statistical analysis was by the likelihood ratio and paired Student's t tests (P <.05). RESULTS: Fetal myoblasts proliferated at faster rates than expected from neonatal cells. Cell attachment was enhanced in the PGA/PLLA matrix and collagen hydrogel when compared with the PGA/P4HB composite. Necrosis was observed at the center of all constructs, directly proportional to cell seeding density and time in the bioreactor. CONCLUSIONS: Fetal myoblasts can be expanded rapidly in culture and attach well to PGA/PLLA, as well as collagen hydrogel but less optimally to PGA/P4HB. Excessive cell seeding density and bioreactor time may worsen final construct architecture. These findings should be considered during in vivo trials of muscle replacement by engineered fetal constructs. PMID- 14523820 TI - Incidence of medical error and adverse outcomes on a pediatric general surgery service. AB - BACKGROUND/PURPOSE: The Institute of Medicine has identified medical error as a leading cause of death and injury, with deaths resulting from medical error exceeding those caused by motor vehicle collisions, breast cancer, or AIDS. The authors examined the incidence and sources of medical error in relation to adverse events on a pediatric general surgery service. METHODS: All intensive care unit (ICU) and ward admissions to 2 staff pediatric general surgeons during a 1-month period were identified prospectively and in-patient care was reviewed daily by a 3-person panel consisting of a staff surgeon, a surgical fellow, and a nonmedical observer. Medical errors, identified through daily patient encounters, nursing rounds, medical rounds, and chart examinations, were evaluated based on type, hospital setting, personnel involved, and outcome. Adverse outcomes were evaluated based on type and contributing factors. RESULTS: Our study group included 64 patients. A total of 108 errors were identified; 28% of these errors resulted in adverse outcomes. One or more medical errors were identified in the care of two thirds of patients (43 of 64), with medical error contributing to adverse outcomes in one third of patients (21 of 64). Errors occurred most frequently in communication, postoperative monitoring and care, and diagnosis, with errors in postoperative care and diagnosis having the highest likelihood of resulting in an adverse outcome. Seventy-four adverse outcomes were identified in 31 patients; 35 (47%) of these outcomes, occurring in 21 patients, were attributable to medical error. The most common adverse outcomes identified were additional nonoperative procedures, of which, 92% resulted from medical error. There were no deaths. CONCLUSIONS: Medical error occurs in more than one half of hospital admissions on a general pediatric surgery service and contributes to a substantial number of adverse outcomes. PMID- 14523819 TI - Benefit of surgical resection of invasive pulmonary aspergillosis in pediatric patients undergoing treatment for malignancies and immunodeficiency syndromes. AB - PURPOSE: The aim of this study was to evaluate the outcome of children undergoing treatment for malignancy and immunodeficiency syndromes in whom invasive pulmonary aspergillosis (IPA) developed. METHODS: The authors reviewed the medical records of all patients treated at their institution from January 1990 to August 1999 for culture-proven pulmonary aspergillus infection. RESULTS: Among the 43 patients studied, the median age at the time of diagnosis of IPA was 13.1 years. The most common primary diagnoses were acute myelogenous leukemia (n = 18) and acute lymphoblastic leukemia (n = 14); 27 patients (63%) had received a bone marrow transplant (BMT). Of the 18 patients who underwent surgical intervention for IPA, 14 (78%) had one operation, whereas the remaining 4 patients had 2. The 4 patients alive at the time this report was written had undergone surgical intervention 2, 10, 23, and 44 months previously respectively. Surgical resection of the involved lung parenchyma was significantly prognostic for survival (P <.001). Other factors that influenced outcome were the extent of pulmonary invasion, steroid use, and the timing of bone marrow transplantation (BMT) in regard to the diagnosis of IPA. CONCLUSIONS: The overall mortality rate of children treated for malignancies and immunodeficiency syndromes in who IPA develops remains high, and antifungal therapy alone may not be curative. Surgical resection may provide a small but possibly the only chance for survival. Therefore, we would advocate for resection of all involved tissue, even if it requires reoperation. PMID- 14523821 TI - Gastric smooth muscle contractility changes in the esophageal atresia rat model: an in vitro study. AB - PURPOSE: The aim of the study was to investigate the gastric smooth muscle reactivity in the Adriamycin-induced esophageal atresia (EA) rat model. METHODS: The fetuses were divided into 3 groups. The control group was exposed to saline. The second group was comprised of fetuses that were exposed to Adriamycin but did not have EA (Adriamycin-no-EA group). The third group was comprised of fetuses that were exposed to Adriamycin and had EA (Adriamycin-EA group). Gastric fundus strips were studied in vitro for their contractile response to receptor activation in the 3 groups. RESULTS: Contractile responses of gastric smooth muscle to carbachol and KCl were increased in the Adriamycin-EA group compared with the Adriamycin-no-EA group. Also serotonin-induced contractile response in the Adriamycin-EA group decreased compared with the Adriamycin-no-EA group. Relaxation of gastric smooth muscle strips to isoproterenol was comparably unaffected in the Adriamycin-EA and Adriamycin-no-EA groups. Likewise, no change in the response to agonist studies was observed between the control and Adriamycin-no-EA groups. The relaxant response to papaverine was not different in the 3 groups. CONCLUSIONS: This study found changes of receptor-dependent and receptor-independent contraction of the gastric fundus smooth muscle in the fetuses with EA. Therefore, impaired contractile responses may be, at least in part, a contributing factor in the abnormal gastric motility seen in EA. PMID- 14523822 TI - Adopt a wait-and-see attitude for patent processus vaginalis in neonates. AB - PURPOSE: The purpose of this paper is to describe the ultrasonographic findings of the patent processus vaginalis (PPV) in neonates. METHODS: The patency of the processus vaginalis was examined by ultrasonography in 117 neonates. The ultrasonographic findings, with increment and decrement of the intraabdominal pressure, were categorized into 6 types as follows: type I, the intraabdominal organ is observed; type II, cystic PPV; type III, the PPV is widened with abdominal pressure increment, the length is > or =20 mm; type IV, the PPV contains moving fluid without PPV widening; type V, the PPV is widened with abdominal pressure increment, the length is less than 20 mm; type VI, others. The authors we regarded types I to IV as PPV with inguinal hernia. RESULTS: Twenty two of 40 neonates with a birth weight under 2,500 g had PPV, including 8 with type I. Twenty of 37 premature neonates 22 to 37 gestational weeks had PPV, including 8 with type I. Eighty-one percent (13 of 16) of PPV in low-birth-weight neonates and 91% (10 of 11) in premature neonates closed spontaneously. The median ages at the time of spontaneous regression of PPV were 242 days in low birth weight neonates and 262 days in premature neonates. CONCLUSIONS: Most premature or low-birth-weight neonates with PPV regress spontaneously. The inguinal hernia in neonates (especially in premature or low-birth-weight neonates) should be observed until at least 9 months of age without attempting hernia repair. PMID- 14523823 TI - Lower prevalence of IL-4 receptor alpha-chain gene G variant in very-low-birth weight infants with necrotizing enterocolitis. AB - BACKGROUND/PURPOSE: Altered production of immunoregulatory cytokines is associated with the development of necrotizing enterocolitis (NEC) in preterm very low-birth-weight (VLBW) infants. According to data obtained in adults, functional genetic polymorphisms influence cytokine production capacity. The aim of this study was to evaluate whether functional polymorphisms of interleukin (IL)-1beta, IL-4 receptor alpha-chain (IL-4ra), IL-6, and IL-10 genes might be associated with the risk of NEC in VLBW infants. METHODS: Dried blood spot samples of 46 VLBW infants with NEC were analyzed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) methods. Samples from 90 VLBW infants without NEC were used as controls. RESULTS: Infants with NEC carried the mutant variant of IL-4ra less frequently than controls (0.125 v 0.224; P <.05) even after adjustment for risk factors of NEC. No significant differences were found in the allelic frequencies of IL-1beta, IL-6, and IL-10 genes between NEC and control infants. CONCLUSIONS: Carrier state of IL-4ra mutant allele might be associated with lower risk of NEC in VLBW infants. This genetic variant is associated with enhanced IL-4 effect. IL-4 is a major regulator of Th1-Th2 shift. The authors hypothesize that infants carrying the IL 4ra mutant allele might have Th2 skewness that might defend against the development of NEC. PMID- 14523824 TI - Bariatric surgery in adolescence. AB - BACKGROUND/PURPOSE: Surgical treatment for morbid obesity is relatively contraindicated in patients less than 18 years of age. However, on some occasions, there is extreme obesity in this age group that does not respond to nonsurgical treatment. The aim of this study was to evaluate the surgical management of severe morbid obesity in adolescents. METHODS: During a 4-year period, the authors assessed 11 adolescent patients with severe morbid obesity. All patients underwent extensive preoperative evaluation including thorough psychological evaluation together with their families. Laparoscopic adjustable gastric banding (LAGB) was performed in all patients. Patients underwent follow up for a mean of 23 months. RESULTS: The mean age of the 11 children in this study was 15.7 years (range, 11 to 17 years). Associated conditions included heart failure and pulmonary hypertension in one patient, amenorrhea in 2 patients, and gallstones in 1 patient. Mean preoperative body mass index (BMI) was 46.4 kg/m2 (range, 38 to 56.6). There were no operative complications. Over a mean follow-up period of 23 months (range, 6 to 36 months), the mean BMI dropped from 46.6 to 32.1 kg/m2 with marked improvement in medical conditions and general psychologic well being. No late complications developed in any patient. CONCLUSIONS: Children are routinely excluded from bariatric surgery programs because of the difficulties involved in psychologically and cognitively preparing this population for surgery. However, extreme morbid obesity rarely responds to nonsurgical therapy for any extended period. This select population will benefit from bariatric surgery if an effort is made to properly prepare patients, together with their families, for the postoperative change in lifestyle and body image. PMID- 14523825 TI - Deficient motor innervation of the sphincter mechanism in fetal rats with anorectal malformation: a quantitative study by fluorogold retrograde tracing. AB - BACKGROUND/PURPOSE: Deficiency of motoneuron innervation to the sphincter mechanism has been described in patients with anorectal malformation. Whether this event is primary or secondary remains unclear. METHODS: The authors quantified the motoneuron innervation of the sphincter mechanism by Fluorogold (FG) retrograde tracing experiment in fetal rats with anorectal malformation. Anorectal malformation was induced in rat fetuses by ethylenethiourea (ETU). Serial longitudinal sections encompassing the whole width of lumbosacral spinal cord were examined. The number of FG-labelled motoneurons were scored and compared between male fetuses with or without malformation in the ETU-fed group and normal controls. RESULTS: The number of FG-labelled motoneurons in the fetuses without defect, with imperforate anus (IA), with neural tube anomalies (NTA), with combined IA and NTA, and normal controls were determined to be (mean +/- SEM) 109.13 +/- 37.88, 55.05 +/- 25.85, 48.20 +/- 30.34, 54.43 +/- 28.55, and 135.22 +/- 28.78, respectively. FG-labelled motoneurons in the fetuses with IA, NTA, and combined IA and NTA are significantly fewer than that in fetuses without defects (P <.05) and in normal controls (P <.005). CONCLUSIONS: These findings suggest that defective motoneuron innervation to the sphincter mechanism is a primary anomaly that coexists with the alimentary tract anomaly in anorectal malformation during fetal development. The intrinsic neural deficiency is an important factor likely to contribute to poor postoperative anorectal function despite surgical correction of anorectal malformation. PMID- 14523826 TI - An infrequent cause of misdiagnosis in esophageal atresia. AB - Inability to pass an oral tube in infants with esophageal atresia often leads to the diagnosis of this anomaly. This report describes an infant with a delay in diagnosis resulting from initial passage of an oral tube through a distal tracheoesophageal fistula into the stomach. PMID- 14523827 TI - Polycystic ovaries, precocious puberty and acquired hypothyroidism: The Van Wyk and Grumbach syndrome. AB - Long-standing acquired hypothyroidism can rarely be associated with precocious puberty and polycystic ovaries. The authors report such a case, which responded to the simple replacement of thyroid hormone. It is important to look for hypothyroidism in girls with ovarian masses and precocious puberty to avoid surgery on the ovaries. PMID- 14523828 TI - Giant pseudoaneurysm of the internal carotid artery causing upper airway obstruction in a 10-month-old infant treated by endovascular occlusion and surgical drainage. AB - A rare case of acute upper airway obstruction caused by a giant compressing pseudoaneurysm of the left internal carotid artery is presented. The pseudoaneurysm developed one month after a deep neck space infection, and an emergency tracheal intubation was necessary. The treatment consisted of endovascular occlusion of the left internal carotid artery and surgical drainage of the pseudoaneurysm with complete resolution of the symptoms. PMID- 14523829 TI - Duodenal atresia associated with proximal jejunal perforations: a case report and review of the literature. AB - Duodenal atresia is associated with Down's syndrome, malrotation, and congenital cardiac defects. Idiopathic intestinal perforations in a newborn, which are not associated with necrotizing enterocolitis (NEC), have been described. The authors report on a full-term neonate who had multiple perforations of the proximal jejunum distal to duodenal atresia. To the best of the authors knowledge, the combination of idiopathic intestinal perforation and duodenal atresia has not been reported previously. PMID- 14523830 TI - Fishbone ingestion: two cases of late presentation as pediatric emergencies. AB - The authors report 2 cases of pediatric emergencies caused by fishbone ingestion. In the first case, 2 fishbones within the sac of an inguinal hernia were detected during an emergency operation for suspected testicular torsion; in the second case, a big fishbone was found in the omentum covering an inflammed appendix during an emergency appendicectomy. In some cases this three foreign bodies can be detected with a simple abdominal X-ray; the relationship between the fish species involved and the diagnosis is discussed. PMID- 14523831 TI - Heterotopic gastric mucosa in a duplicate gallbladder. AB - A 7-year-old girl presented with recurrent acute pancreatitis. Detailed investigations, including endoscopic retrograde cholangiography, suggested a poorly defined biliary tract abnormality. At laparotomy, this was discovered to be a duplicate gallbladder each with a separate cystic duct but contained within a single serosal envelope. Both gallbladders were removed, and histologic examination found the inferior organ to be lined by heterotopic fundic-type gastric mucosa. Despite the absence of any remaining structural biliary abnormality and no evidence of residual ectopic gastric mucosa, the patient experienced a few further episodes of self-limiting mild acute pancreatitis during the following 3 years. The presence of heterotopic gastric mucosa in a duplicate gallbladder has not been described previously. PMID- 14523832 TI - Perirenal tailgut cyst (cystic hamartoma). AB - Tailgut cysts are rare congenital multicystic lesions preferentially occurring in the retrorectal space. They are assumed to arise from the remnant of embryonic tailgut and occasionally exhibit malignant transformation. The authors herein describe the clinical, radiologic, and pathologic features of an unusual perirenal tailgut cyst with an emphasis on its histogenesis and distinction from other intraabdominal cystic lesions. PMID- 14523833 TI - Severe acute necrotizing pancreatitis associated with lipoprotein lipase deficiency in childhood. AB - An 11-year-old girl with lipoprotein lipase deficiency experienced recurring episodes of abdominal pain. She initially underwent appendectomy for suspected appendicitis; however, the appendix was normal. Pancreatitis was subsequently identified as the cause of her pain. PMID- 14523834 TI - Cholecystoenteric fistula: a rare complication of necrotizing enterocolitis. AB - An infant treated with necrotizing enterocolitis, had a cholecystoenteric fistula, which was found incidentally on routine contrast study of his intestinal tract before closure of his ileostomy. To the best of the author's knowledge this complication has not been reported before. PMID- 14523835 TI - Hirschprung's disease and imperforate anus in Pallister-Hall syndrome: a new association. AB - Hirschprung's disease and imperforate anus are described concurrently in a newborn with Pallister-Hall syndrome as well as the difficulties in making this diagnosis. Awareness of this new association should prompt the exclusion of Hirschprung's disease before repair of imperforate anus in infants with Pallister Hall syndrome. The known genetic parallels between these conditions is discussed briefly in terms of etiology. PMID- 14523836 TI - Supradiaphragmatic accessory liver: a rare cause of respiratory distress in a neonate. AB - The authors describe a case of supradiaphragmatic accessory liver seen on prenatal ultrasound examinations, mimicking fetal hydrothorax and causing life threatening respiratory failure after birth. The accessory liver with a transdiaphragmatic pedicle to the caudate lobe of the normal liver was found at throracotomy after failure to expand the lung by chest drainage. The "hydrothorax" turned out to be multilocular mesothelial cyst collection surrounding the ectopic liver. This is the first report of the development of a thoracic accessory liver followed by obstetric ultrasound examinations. PMID- 14523869 TI - Submuscular bar, multiple pericoastal bar fixation, bilateral thoracoscopy, modified Nuss repair in adolescents. PMID- 14523837 TI - A rapidly growing flank mass in an infant. PMID- 14523871 TI - Survival after CCAM associated with ascites: a report of a case and review of the literature. AB - A congenital cystic adenomatoid malformation (CCAM) is a congenital lung lesion that, when associated with hydrops, is thought to have a dismal prognosis without fetal intervention. The authors report a case of a fetus with a CCAM associated with ascites as the only manifestation of hydrops, which was diagnosed at 19 weeks gestation. The CCAM regressed without intervention between 23 and 29 weeks of gestation with resolution of the ascites. The baby was born asymptomatic at term, and the infant underwent elective resection of the CCAM at 1 year of age. A review of the literature from 1980 to 2000 yielded 9 other cases of spontaneous resolution of hydrops associated with CCAM, suggesting that although rare, this outcome is a possibility in fetuses diagnosed with the condition. Because most fetuses diagnosed with hydrops undergo termination of pregnancy, the true incidence of hydrops resolution is potentially higher than predicted. This has important implications for prenatal counseling particularly in centers in which fetal intervention is not available. PMID- 14523872 TI - Anorectal malformation with recto-perineal fistula: case report and clues to diagnosis. AB - Rectoperineal fistula is an unusual form of anorectal malformation in a boy. The authors report on a patient with this anomaly and the implications in terms of difficulty in diagnosis and neonatal management. PMID- 14523873 TI - Serous cystadenoma of the pancreas in a child. AB - Pancreatic tumors are very rare in children. Until now, approximately 150 cases have been reported in the English-language medical literature. Only 4 of them represented serous cystadenoma. Although the tumor is not potentially malignant, all patients required partial pancreatic resection. The authors present a 4-year old girl with a short history of abdominal pain and vomiting. Sonography and computed tomography showed a pancreatic mass. The mass was proved to be a serous cystadenoma, and resection of the tumor was performed. Clinical discussion with review of the literature is presented. PMID- 14523874 TI - Strongyloides appendicitis: unusual etiology in two siblings with chronic abdominal pain. AB - Appendicitis is one of the most common causes of acute surgical disease in children and young adults. Parasites, however, are one of the uncommon etiologies. An 8-year-old girl and her 7-year-old sister presented with more than 2 months of chronic abdominal pain that became worse over a 1-week period before presentation. The 2 sisters presented 1 month apart. Both had similar symptomatology and physical examination findings. At operation, the surgical findings included an inflamed appendix with a cross section of the parasite Strongyloides. Strongyloides appendicitis has occurred almost exclusively in areas endemic to the parasite. Its environment is more common outside the United States but occasionally is seen in the Southeast region and in institutionalized individuals. The presentation of acute exacerbation of chronic abdominal pain coupled with the pathologic finding of Strongyloides in an acutely inflamed appendix, should alert the clinician of other possible cases. This increased index of suspicion will allow more prompt diagnosis and help avoid the morbidity of delayed operation. PMID- 14523875 TI - Small bowel obstruction caused by congenital mesocolic hernia: case report. AB - Transmesocolic hernias are extremely rare. Their exact incidence is still unknown. A strangulated hernia through a mesocolic opening is a rare operative finding. Preoperative diagnosis still is difficult in spite of imaging techniques currently available. This is the case of a 4-month-old boy with transmesocolic internal hernia and coincident intestinal malrotation and volvulus of small bowel. PMID- 14523876 TI - Triorchidism and testicular torsion in a child. AB - Triorchidism is a rare congenital abnormality of the genitourinary tract. The first surgical case was described in 1895 by Lane. The authors report an interesting case of 2 separate right testes found when a left twisted testis was removed. The generally endorsed necessity to remove the duplicate testis in triorchidism is discussed. This procedure has been recommended because of the risk of torsion. PMID- 14523877 TI - Continent catheterizable urinary conduit constructed from defunctionalized colon. AB - The authors describe a technique for construction of a continent catheterizable stoma from distal defunctionalized colon in a patient with imperforate anus, urethral atresia, and sacral agenesis. PMID- 14523878 TI - Bloody diarrhea caused by a needle in the pancreatic head. AB - Bloody diarrhea in adolescents is not uncommon polyposis, inflammatory bowel disease, and infections are the usual causes. A 13-year-old boy with a metallic foreign body in the head of the pancreas presented with bloody diarrhea. The clinical problems and surgical approach are discussed. PMID- 14523879 TI - Fluoroscopy-guided laparoscopy in the management of intraabdominal foreign body. AB - Appendicitis or perforation as a result of intraluminal foreign body is uncommon. The method of diagnosis and proper approach for an elective appendectomy in an otherwise silent intraluminal foreign body is under dispute. A laparoscopic exploration under intraoperative fluoroscopy guidance was performed in a mentally retarded 9-year-old boy with asymptomatic foreign bodies in the right lower abdominal quadrant. The authors recommend laparoscopic appendectomy, assisted by intraoperative fluoroscopy for patients with pointed foreign bodies in the appendix. PMID- 14523880 TI - Preoperative embolization as an adjunct to the operative management of mediastinal Castleman disease. AB - Castleman disease usually presents in children as a localized mass with prominent feeding vessels. The mainstay of treatment of Castleman disease is surgical resection; historically, resection is associated with excessive blood loss. These tumors are well known to have large feeding vessels and, thus, are amenable to preoperative arteriography with embolization. The authors present a case of Castleman disease treated with preoperative embolization as an adjunct to operative management. PMID- 14523881 TI - 2002 annual report of the American Association of Poison Control Centers Toxic Exposure Surveillance System. PMID- 14523882 TI - Use of dosage as a triage guideline for unintentional cyclic antidepressant (UCA) ingestions in children. AB - Triage guidelines for unintentional cyclic antidepressant (UCA) ingestions vary widely, with limited supportive evidence. All records of UCA ingestion reported to 4 certified regional poison centers were evaluated for the years 1998 through 2000. Inclusion criteria included age 5 mg/kg. The majority of UCA ingestions produced limited or no symptomatology. In this series, all children with ingestions of <5 mg/kg developed no or minor effects. Home monitoring might be appropriate in such cases. PMID- 14523883 TI - Predictors of delay in presentation to the ED in patients with suspected acute coronary syndromes. AB - Delays in seeking medical attention for patients with acute coronary syndromes (ACS) preclude early application of life-saving treatment and diminish efficacy. Previous studies suggest 3-hour delays between onset of symptoms and ED arrival in patients with typical presentations of acute myocardial infarction (AMI). A prospective observational study was conducted in an urban ED measuring lag time (LT) among adults presenting within 48 hours of onset of symptoms suggestive of ACS. Univariate and multiple regression analyses were performed on 5 predictors: age, sex, symptoms at presentation, and 2 different outcomes (AMI and ACS). Three hundred seventy-four patients were enrolled. Mean age was 63 years with 38% 70 years or older. Seventy-three percent of all patients with suspected ACS presented with chest pain, 27% with atypical symptoms. Overall mean LT was 8.7 hours (standard deviation 11). In subgroup analysis, patients aged >/=70 years were more likely to have LTs >12 hours (29% vs. 19% P =.043) and patients without chest pain had longer mean LTs (11.6 vs. 7.6 hours, P =.01). Delay in ED presentation is group specific. Advanced age and patients with atypical symptoms are predictive of longer LTs. Contrary to previously published data, patients with symptoms suspicious for ACS can delay an average of 9 hours, which might alter current thinking in the prevention and care of these patients. PMID- 14523884 TI - 70-mph speed limit and motor vehicular fatalities on interstate highways. AB - In May 1996, Alabama increased the rural interstate highways speed limit to 70 mph. This study was conducted to determine if the faster speed limit had any influence on the number of motor vehicular crash (MVC) deaths. The null hypothesis was that no significant change occurred in the number of deaths since the faster speed was introduced. Using MVC fatality data collected by the Alabama Department of Public Safety and published by the state of Alabama from 1984 to 1999, a time-series study designed was used to examine the number of MVC deaths before and after the speed limit change. The trend of MVC deaths was examined for both rural and urban interstate highways and those occurring on Federal and state highways. There were 174 interstate MVC deaths in 1997 and 165 in 1999; both figures represent a significant increase (P <.05) from the trend of previous years' results. State and Federal highways demonstrated no significant change. By 1998 the number of interstate highway MVC deaths dropped to 114, falling back within the expected trend established by the results of the previous years. The number of rural interstate MVC deaths was found to be significant in 1997 and 1999. The faster speed limit was associated with an increase in MVC deaths for the first year after the speed limit increase. The decline in the number of MVC deaths for 1998 was unanticipated. There are many other factors that need to be evaluated before determining the cause and effect that higher speed limits have on MVC fatalities. PMID- 14523885 TI - Pain medication use before ED arrival. AB - The objective of this study was to determine the frequency and types of pain medications taken before ED arrival based on pain intensity, duration of pain, chief complaint, gender, age, and race. A convenience sample of patients in pain was enrolled in this university hospital-based prospective, observational study. A total of 1233 patients were enrolled. Five hundred thirty-nine of 1233 (44%) patients took pain medication before arrival. Two hundred three (38%) took ibuprofen, 147 of 539 (27%) took oral opioids, and 135 of 539 (25%) took acetaminophen, which were the most frequently used medications. Severity of pain, age, duration of pain, and chief complaint were associated (chi-squared P <.05) with variations in prior medication use. Race and gender were not associated (chi squared P >.05) with differences in medication use before arrival. Many patients (44%) take medication before arrival in the ED. Age, severity and duration of pain, as well as chief complaint are associated with differences in frequency of self-administered medication. PMID- 14523886 TI - Efficacy of alteplase thrombolysis for ED treatment of pulmonary embolism with shock. AB - Our objective was to assess efficacy and tolerance of thrombolysis using 0.6 mg/kg of Alteplase in patients with massive pulmonary embolism defined as the association of a pulmonary embolism with shock. We retrospectively included 21 patients presenting with a massive pulmonary embolism confirmed by either scintigraphy or spiral computed tomography. Patients were treated on the basis of a standard rationale followed by thrombolysis with 0.6 mg/kg Alteplase over a period of 15 minutes. Hospital mortality, vital signs before and 2 hours after thrombolysis, and incidence of hemorrhagic events were recorded. Five patients (23.8%) died, 4 of these deaths occurred during the first 4 hours after hospital admission. Systolic and diastolic blood pressure (Sp02) were significantly improved 2 hours after the beginning of thrombolysis. Five minor hemorrhagic events occurred. This study demonstrates that for patients with pulmonary embolism and shock, a bolus treatment with Alteplase is potentially effective and well tolerated. PMID- 14523887 TI - The effect of IM ketorolac tromethamine on bleeding time: a prospective, interventional, controlled study. AB - Opiates, although effective analgesics, have significant adverse side effects. Ketorolac, the only parental nonsteroidal antiinflammatory drug available for use in the United States does not cause significant respiratory depression or hypotension, but it is a reversible inhibitor of platelet aggregation with a theoretical increased bleeding risk, which limits its use. The objective of this study was to determine the effect of a single intramuscular dose of 60 mg ketorolac on 4-hour bleeding times in healthy volunteers. This was a prospective, paired, unblinded, before-and-after interventional study performed in a suburban university-based EM residency training program. Subjects were 20 healthy volunteer EM residents. Standard Ivy bleeding times were measured before and 4 hours after intramuscular administration of 60 mg ketorolac. Before-and-after bleeding times were compared using a paired t-test. The study had 90% power to detect an effect size of 0.5. The subjects' mean age was 31.6 and 7 (35%) were females. Bleeding time was increased from a mean baseline time of 3 minutes 34 seconds (+/- 1 min 20 sec) to a mean 4-hour postinjection time of 5 minutes 20 seconds (+/- 3 min 8 sec). The mean prolongation of bleeding time was 1 minute 46 seconds (50% increase with 95% confidence interval, 25%-75%). There were no adverse events. A standard intramuscular dose of 60 mg ketorolac resulted in prolongation of the bleeding time in healthy volunteers. The clinical significance of this prolongation in patients is unclear. PMID- 14523888 TI - The significance of routine serum magnesium determination in the ED. AB - To determine the significance of routine serum magnesium determination in the ED, a retrospective analysis was done on 253 patients in whom serum magnesium levels were examined in the department of EM. Mild, moderate, and severe hypomagnesemia was found in 19.5%, 9.1%, and 2.5% of the study population, respectively. Univariate analysis showed that pregnancy and pregnancy-related conditions, diabetes mellitus, gender, renal function, infectious diseases, and the presence of ischemic heart disease were associated with low serum magnesium levels. In multivariate analysis, only pregnancy and pregnancy-related conditions (odds ratio [OR], 4.39; 95% confidence interval [CI], 1.2-15.3) and diabetes mellitus (OR, 9.04, 95% CI, 3.3-24.75) were significantly associated with low serum magnesium levels. The number of patients admitted to the hospital and the number of patients who died was not statistically significantly different between the groups of patients with normal serum magnesium levels as compared with those with low serum magnesium levels. Although hypomagnesemia was found in 31% of the study population, its clinical significance is not clear given the absence of hypomagnesemia-related complications and death. PMID- 14523889 TI - Salicylism and herbal medicine. PMID- 14523890 TI - ST segment monitoring as a predictor of reperfusion after blood transfusion in a patient with acute non-Q-wave myocardial infarction. PMID- 14523894 TI - [Clinical research and its evaluation]. AB - Biomedical research may be defined as "basic" or "applied." Applied research may be "pre-clinical" or "clinical." The final products of research are publications in peer-review system journals and patents. The measurement of quality in biomedical research is performed by using many parameters, such as impact factor, citations, and number of granted projects. In the United States and some countries in Europe, the evaluation of research in public and private institutions is done by a council of faculty members or by accredited agencies. In Italy, mass media commissions the measurement of quality in biomedical research to non-accredited institutions, as was done recently by Corriere della Sera-Salute. PMID- 14523895 TI - [Composition of proteinuria in primary glomerulonephritides: association with tubolo-interstitial damage, outcome and response to therapy]. AB - Experimental and clinical studies show that proteinuria is an independent risk factor for the progression of chronic glomerular diseases and is associated with the extent of tubulo-interstitial damage. The accumulation of proteins in tubular cells induces the increased expression of a variety of inflammatory and fibrogenic cytokines, with the consequent development of interstitial inflammation, a proliferation of fibroblasts, the increased production of extracellular matrix, and the formation of interstitial fibrosis. Laboratory methods, such as immunonephelometry, can easily evaluate the glomerular component of proteinuria, due to the alteration of the structural integrity of the glomerular capillary wall. This alteration allows the tubular lumen to pass proteins of high and middle molecular weight (HMW and MMW proteins: IgM, alpha2 macroglobulin, IgG, transferrin, albumin). Using the same method and SDS-PAGE, it is possible to evaluate those tubular components of proteinuria that are composed of low molecular weight (LMW) proteins, such as alpha1-microglobulin (alpha1m) and beta2-microglobulin (beta2m), whose reabsorption by tubular cells-almost complete in physiological conditions-is impaired in pathological conditions. Recent studies clarified some aspects of the relationships between the components of proteinuria, histological lesions, prediction of outcome, and response to therapy. The extent of tubulo-interstitial damage is correlated with selectivity of proteinuria and IgG excretion, suggesting a possible tubulo-toxic role for IgG or for some other protein of similar molecular weight. The tubulo-interstitial lesions are also correlated with the excretion of LMW proteins, due to their impaired reabsorption. The remission of nephrotic syndrome, not predicted by the amount of proteinuria, is highly predicted by the selectivity index or IgG excretion in membranous glomerulonephritis (MGN) and focal segmental glomerulosclerosis (FSGS). Progression to chronic renal failure is better predicted both by the glomerular component (selectivity index, IgG excretion) and by the tubular component of proteinuria (alpha1m, beta1m, LMW proteins), than by 24-hour proteinuria. The response to therapy in MGN and FSGS is dependent on the excretion of IgG and alpha1m. In conclusion the composition of proteinuria can easily be assessed using automated methods, and it is useful to evaluate the relationship of proteinuria with histological lesions to predict the functional outcome and response to therapy in primary glomerulonephritis. PMID- 14523896 TI - [A genetic viewpoint of focal glomerular sclerosis: fom genes to glomerular pathophysiology [corrected]]. AB - Recent studies of Mendelian disease have begun to clarify the clinical spectrum of the group of disorders that make up familial, focal segmental glomerulosclerosis (FSGS) and nephrotic syndromes. In familial forms of focal segmental glomerulosclerosis (FSGS), both autosomal recessive and dominant inheritance patterns have been reported. At least three genes have been identified which, when defective, cause familial FSGS or nephrosis: the NPHS1 gene, encoding nephrin; the NPHS2 gene, encoding podocin; and the ACTN4 gene, encoding a-actinin-4. Because the majority of FSGS cases occur as sporadic disease, the recently described mutations in the NPHS2 gene "in approximately 25 percent of cases of apparently sporadic, steroid-resistant FSGS in children" have claimed great interest. The applicability of these observations to adults, including the possible importance of the nephrin and alpha-actinin-4 genes in the sporadic disease, remain to be determined. Finally, the mechanisms of podocyte damage and the molecular basis of glomerulosclerosis are reviewed. PMID- 14523897 TI - [Clinical management of the patient affected with acute renal failure (ARF) secondary to volume depletion]. AB - Most cases of ARF are secondary to volume depletion. In the literature, very few scientific publications address the problem of what to do when confronted with such a patient. As regarding the diagnosis of hypovolemia, an accurate history and physical examination can help to determine both the presence and etiology of volume depletion; postural hypotension (decrement in systolic blood pressure of more than 20 mmHg after standing from the supine position), associated with a pulse increment of 30 beats/min or more and dizziness are specific symptoms of hypovolemia. Laboratory indices are useful to diagnose volume depletion, but their interpretation is not simple, and they may not be available in the non nephrologic environment. Fluid replacement therapy in hypovolemia is largely dependent upon the type of fluid that has been lost and concurrent electrolytic and acid-base disorders. Patients with hypernatremia and volume depletion should receive mild hypotonic solutions, whereas those with hyponatremia and hypovolemia should receive mild hypertonic solutions. The entity of reinfusion depends on daily losses. Conversely, monitoring of body weight can be considered an adequate index of fluid balance. Concerning the treatment of ARF, the use of loop diuretics in the early phases of pre-renal ARF decrease oxygen consumption in the tubular cells by inhibiting transcellular sodium transport, therefore preventing or limiting ischemic cell injury. The use of loop diuretics should also be evaluated in intermediate syndrome and ischemic NTA where diuretics can, respectively, reduce renal ischemia and convert oliguric ARF into the non oliguric form. PMID- 14523899 TI - [Collaborative study on peritoneal dialysis (PD) as first dialysis treatment in an Italian region: 1994-2000]. AB - BACKGROUND: In Lazio, only about 5% of uremic patients are on peritoneal dialysis (PD). The present study focuses on the parameters of PD selection, the treatment schedules, and the clinical outcomes of PD patients in the nine public facilities offering a PD program. A cohort of 249 first-time PD patients, from July 1, 1994 to December 31, 2000, was retrospectively considered. METHODS: For the enrollment of the patients, the Regional Dialysis Registry databank was consulted. On December 31, 2000, a systematic review of patient charts was performed to extract the reasons for the PD choice, details of PD schedule, peritonitis episodes, reasons for drop-out, and patient survival rates. In regard to technique success defined as the probability of having a patient alive on PD-change of modality and death were considered as final events. In regard to patient survival, only death, even in the first 2 months after a shift to hemodialysis, was considered the end point. RESULT: The main PD selection reasons were patient and/or nephrologist preference in 90% of cases. One-hundred eighty-nine patients (76%) had been started on CAPD. During the follow-up, 38.2% dialysis schedules had been modified at least once. At the end of follow-up, 41.2% patients were on APD. The peritonitis rate was one episode per 30 patient-months (1 per 27 patient-months in CAPD; 1 per 37 patient-months in APD; p = 0.08). The technique success rate was 66.3% after 2 years and 49.8% after 3 years. The patient survival rate was 81.1% after 2 years and 68.7% after 3 years. CONCLUSIONS: Patients chose PD as a first dialysis treatment mainly because of reasons unrelated to their clinical status. The technique's success, patient mortality rates, and the peritonitis rate do not explain the low PD diffusion in the region. The peritonitis rate meets the target criteria for excellence recommended by the Italian Society of Nephrology. The observed outcomes may have been favored by the selection of motivated patients and by the increased use of APD. PMID- 14523898 TI - [Hepatocyte growth factor (HGF) reduces the expression of profibrotic factors in human isolated glomeruli]. AB - BACKGROUND: The imbalance between the synthesis and degradation of the mesangial matrix causes glomerulosclerosis and ultimately leads to chronic renal failure. HGF is a pleiotropic cytokine involved in angiogenesis, morphogenesis, organogenesis, and bone remodeling. Recently, we and other investigators have shown that HGF has a central role in the recovery of acute renal failure. Furthermore, HGF treatment halts the progression of kidney disease in a murine model of chronic renal failure. The aim of the present study was to evaluate the effect of HGF on the mRNA levels of molecules involved in the extracellular matrix turnover and of the c-met receptor in isolated human glomeruli. METHODS: Human glomeruli were isolated by microdissection from donor kidney biopsies just before transplantation. Glomeruli were extensively washed and incubated with culture media containing HGF (50 ng/mL) for 24 h at 37 C. Glomeruli incubated without HGF were used as controls. After 24 h, glomeruli were washed and freezed and thawed three times. The expression of c-met, (alpha2) IV collagen, TGF-beta, metalloproteases 9 (MMP9), and of the inhibitor of metalloproteases-1, TIMP-1 was evaluated by in situ reverse transcription (RT) and polymerase chain reaction (PCR). beta-actin was used as a housekeeping gene. RESULTS: The (alpha2)IV collagen mRNA level was decreased by HGF in human glomeruli. TGF-beta and TIMP-1 gene expression was markedly reduced by HGF treatment, whereas the expression of MMP-9 and c-met did not change. Under light-microscopic examination, kidney biopsies showed neither glomerular hypercellularity nor mesangial expansion. CONCLUSIONS: HGF treatment reduces the expression of extracellular matrix components and of profibrotic factors in human glomeruli. Our results confirm a protective role of HGF in glomerulosclerosis. PMID- 14523900 TI - [Anti-HLA antibodies in nephropathic patients]. AB - BACKGROUND: The presence of anti-human leukocyte antigen (HLA) alloantibodies in nephropathic patients is due to immunogenic stimuli such as transfusions, pregnancies, and transplantations. These stimuli can be highlighted using a classic aspecific serologic technique, such as complement-dependent cytotoxicity (CDC) or using more recent and specific techniques, such as cytofluorimetrics or enzyme linked immunoabsorbant assay (ELISA). Because the presence of anti-HLA preformed antibodies is linked to the largest incidence of both acute and chronic rejection, it seems appropriate to re-evaluate that data obtained using aspecific classic serological analysis techniques by using the more specific cytofluorimetric technique. To aid in the possible prevention of ant-HLA antibody formation, it is also appropriate to analyze the influence of immunogenic stimuli on the development of these antibodies. METHODS: We studied 116 patients (37 women and 79 men). Anti-HLA antibodies were detected using microlymphotoxic technique after separation of B and T lymphocytes. This separation was obtained using magnetic balls. We used a 30-cell panel. We also used a recent cytofluorimetric test (Flow Pra screening; One Lambda Inc., 21001 Kittridge St., Canoga Park, California, U.S.A.) with a panel of micrograins covered with class I and class II purified antigens. Statistical analysis was performed using chi square analysis or Fischer s exact test. For each test, sensibility, specificity, and positive and negative value were measured. RESULTS: Among 33 patients testing positive using the classic CDC-PRA technique (17 positive for B-lymphocytes and 16 positive for both B and T lymphocytes), using cytometry, 25 were positive for anti-HLA-specific antibodies (10 among the B lymphocyte-positive patients and 15 among the B + T lymphocyte-positive patients). Two patients were shown positive only using the cytofluorimetric method. Of the 27 patients positive at cytometry, 18 were positive for class I and class II, 4 for class I, and 5 for class II. FLOW-PRA screening results were less sensitive and more specific. The results obtained by the two methods are comparable(p<0.0001). The immunogenic stimuli found responsible for immunization were: transfusion in 10 of 25 patients, pregnancies in 3/9 patients, transplant in 4/8 patients, and different immunogenic stimuli in 10/12 patients. The results were not statistically significant (p>0.05). CONCLUSIONS: Data show that positivity for B lymphocytes obtained using CDC-PRA is not always linked to the development of anti-HLA antibodies, whereas positivity for B+T lymphocytes, obtained using CDC -PRA, is often linked to specific antibody development. Immune response is more often directed against class I and II antibodies. The specific detection of HLA antibodies using the cytofluorimetric method allows us to identify patients at risk for rejection, and it suggests that red cells should be filtrated to prevent anti-HLA immunization secondary to transfusion in transplantation candidates. PMID- 14523901 TI - [Historical Archives of Italian Nephrology. Virchow's theories on atherosclerosis and related kidney disease]. AB - This article concerns the development of theories about atherosclerosis and its changing manifestations, some of which, such as renal artery disease and ischemic nephropathy, have been clearly defined only in recent times. An anatomical consequence of atherosclerosis is the stenosis of the renal artery, which opens an unexpected clinical response in blood pressure (nephrovascular hypertension) and in kidney function (ischemic renal disease). Today's historian, no less than the clinician, is called on 'to demand the consideration of basic definitions and nomenclature.' It is impossible to overrate Virchow's theory of artherosclerosis and his great contribution to the concept of artery insudation and thrombosis process. But even Virchow did not expressly stress the concept of atherosclerosis as an autonomic non-inflammatory entity; he called the condition 'chronic endoarteritis deformans.' However, his hypothesis introduced some inflammatory factors that today are the basis of novel atherosclerosis theory. Although Anitschkow introduced the theory of the influence of cholesterol on atherosclerotic development, his theory has been partially contradicted by widespread epidemiological data. Currently, a re-emerging theory on the role of inflammation and infection in atherosclerosis seems to explain many clinical facts taken in small account by other theories. PMID- 14523902 TI - [Recurrence of glomerulonephritis in transplanted kidney]. AB - Recent advances in immunosuppressive therapy have dramatically reduced the incidence of acute rejection, thus improving graft survival. As a result, the importance of recurrence of the original nephropathy as a factor affecting the long-term outcome of the graft has grown considerably The incidence of recurrence increases with the increase in graft survival and can currently be estimated between 6 and 15% 10 years after surgery, with great variability among the different histological types of nephropathy. More than 50% of patients with recurrence experience progressive deterioration of graft function, and recurrence of the nephropathy accounts for long-term graft failure in more than 15% of cases. The original disease should be duly considered in all candidates for renal transplantation to identify patients at higher risk for recurrence and to define those treatment protocols devoted to risk reduction. Finally, the risk of disease recurrence should always be included among the parameters used in evaluating a possible transplantation from living donor. PMID- 14523903 TI - [Medullary sponge kidney (MSK) in association with glomerulonephritis (GNP)]. AB - BACKGROUND: Medullary sponge kidney (MSK) is a congenital, non-hereditary anomaly characterized by dilation of the precalix ducts. It is often associated with other diseases, and its symptoms are linked to frequent complications, such as sepsis of the urinary system and the renal colic of nephrolithiasis. However, MSK is rare in association with glomerulonephritis (GNP), as can be seen from medical literature data that shows only one case (reported in 1974) in which MSK with a concomitant focal sclerosing glomerulonephritis was noted. In this report, we describe a case of MSK that was found after a routine renal echograph and biopsy of a mesangial GNP previously diagnosed at another center. The echographic results showing typical MSK was confirmed by means of RX urography and renal TC. The MSK-glomerulonephritis combination appears to be completely random, because the involved structures (the collecting duct and the glomerule) have different embryologic origin. Therefore, a pathogenic hypothesis common to the two anomalies seems rather unlikely. CONCLUSIONS: Although MSK is a usually benign disease, because a compromise of glomerular filtration that occurs in approximately 10% of cases, proper diagnosis is important to allow prophylactic intervention aimed at the reduction of complications that could reduce overall renal function when associated with other nephropathies. Renal echography is an important diagnostic tool that may reveal more cases of MSK than previously reported. PMID- 14523904 TI - [Acute renal failure caused by VIP-secreting tumor]. AB - A 74-year-old woman had secretory diarrhea, severe metabolic acidosis, hypokalemia, hypovolemia, and acute renal failure caused by a pancreatic vasoactive intestinal polypeptide (VIP)-secreting tumor. Vipoma is a rare neuroendocrine tumor. Morbidity and mortality are related to long-standing dehydration and electrolyte and acid-base disturbance resulting in acute renal failure. Diagnosis requires the documentation of large volumes of secretory diarrhea, elevated VIP plasma levels, and the localization of the VIP-secreting tumor. Metastases are present in 50% of patients at the time of diagnosis. Treatment includes correction of volume, electrolyte, and metabolic abnormalities; CVVH during ARF; pharmacotherapy to decrease gastrointestinal secretion; and surgical resection of the vipoma. PMID- 14523905 TI - [Diabetes mellitus]. AB - Diabetes mellitus is a group of diseases characterized by high levels of blood glucose resulting from defects in insulin production, insulin action, or both. Diabetes is a serious health concern. The number of cases of diabetes mellitus is estimated to grow at a rate of 50% between 2000 and 2010. There are several types of diabetes: type 1 diabetes, type 2 diabetes, gestational diabetes, and other specific types of diabetes. Beta cell dysfunction plays a key role in the physiopathology of diabetes, even when insulin resistance, which is often present in several diabetes-related diseases, is considered among the causes of hyperglycemic type 2 diabetes. The prolonged hyperglycemia that is peculiar to all kind of diabetes has long term complications on several organs and systems. The diagnosis of diabetes is based on the evaluation of glucose plasma levels performed under fasting conditions or two hours after the oral ingestion of 75 grams of glucose. Currently, achieving and maintaining normal plasma levels of glucose are the aims of therapy for both type 1 and type 2 diabetes. Particularly, the therapy for type 1 diabetes is based on the administration of insulin, whereas that of type 2 diabetes changes over the time: diet and physical activity are the first treatments; oral hypoglycemic drugs are used as a second therapeutic step; and the administration of insulin is the last therapeutic option. The principal therapeutic innovation of the past ten years is represented by the tight and flexible control of glucose plasma level obtained by using the insulin analogues produced by recombinant DNA technology. PMID- 14523906 TI - [The incidence, clinical implications, and risk factors of diabetes mellitus]. AB - The incidence of diabetes in kidney transplant recipients is between 4 and 20% in patients previously not affected by this pathology. This difference is partially due to the immunosuppressive therapy administered. The incidence of diabetes is very high during the first quarter after the transplant, and it becomes stable during the following quarters. The presence of diabetes - evaluated by postprandial glycemia and glycated hemoglobin - should be checked quarterly during the first year after the transplant, every six months during the second year, and yearly starting from the third year. The immunosuppressive therapy (calcineurin inhibitors and steroids), familial history, age, race, and weight (BMI) are among the risk factors of diabetes post-transplant. An increased risk of rejection seems to be among the principal consequences of diabetes in transplant recipients. Moreover, these patients are more prone to infections, cardiovascular disease, and the degenerative complications of diabetes. These facts increase the risk of organ insufficiency, morbidity, and mortality. To manage diabetes in transplant recipients it is necessary to identify at-risk patients before the transplantation, thus avoiding complicated and hazardous examinations after the transplant. After the transplantation, the modifiable risk factors, such as the immunosuppressant drugs used and the control of body weight, must be checked. The control of hypertension is important as well. PMID- 14523907 TI - [Calcineurin inhibitors and mechanisms that are responsible for the appearance of post-transplant diabetes mellitus]. AB - Calcineurin (CaN), a calmodulin-dependent heterodimer, is, together with NAF-T, involved in the regulation of the Ca++ pump and in the transmission of the activation signal of the immune response. The CaN inhibitor drugs, such as cyclosporin A (CyA), carried by cyclophillin, and tacrolimus, carried by FK binding protein-12 (FKBP-12), inhibit the binding with the regulatory subunit CaNB. A meta-analysis, comparing tacrolimus with cyclosporin A, has evidenced that tacrolimus significantly increases the diabetes prevalence one year after renal transplant. The diabetogenic effect is due to a direct effect of both drugs on the beta pancreatic cell, in particular on intracellular Ca++ metabolism, which is involved in the insulin secretion and in the reduction of the number of the secretor granules. Some immunological-hystochemical studies, performed on murine pancreas, have evidenced that the FKBP-12 content is higher in beta cells than in alpha cells. This fact allows a high intracellular store of tacrolimus, and a consequently more toxic effect, inside the insulin secreting structures. On the contrary, the low FKBP-12 content of alpha cells involves a higher content of calcineurin and a higher resistance to toxic effects. Finally, an increased incidence of islet cell antibodies (ICA) has been evidenced in patients treated with tacrolimus, as opposed to those treated with CyA. PMID- 14523910 TI - Oligonucleotide-based knockdown technologies: antisense versus RNA interference. AB - The postgenomic era is characterized by an almost intimidating amount of information regarding the sequences and expression of previously unknown genes. In response, researchers have developed an increasing interest in functional studies. At the start of such a study, one may have little more than sequence information and bioinformatic annotation. The next step is to hypothesize a potential role in the context of a cell. Testing of the hypothesis needs to be fast, cheap, and applicable to a large number of genes. Knockdown methods that rely on binding of antisense oligonucleotides to mRNA combined with a subsequent functional assay in cell culture fulfil these requirements: sequence information is sufficient for synthesis of active inhibitors. Depending on the in vitro model chosen, knockdown of gene expression can be achieved with medium or even high throughput. The two most popular methods of knockdown in cell culture are the use of antisense oligonucleotides that rely on ribonuclease H (RNAse H)-dependent cleavage of mRNA, and RNA interference triggered by small double-stranded RNA molecules. Both methods act in a sequence-specific manner and can give efficient knockdown. In both cases, researchers struggle with nonspecific "off-target" effects and the difficulty of site selection. Studies that compare the methods differ in their judgment as to which method is superior. PMID- 14523911 TI - NMR spectroscopy of RNA. AB - NMR spectroscopy is a powerful tool for studying proteins and nucleic acids in solution. This is illustrated by the fact that nearly half of all current RNA structures were determined by using NMR techniques. Information about the structure, dynamics, and interactions with other RNA molecules, proteins, ions, and small ligands can be obtained for RNA molecules up to 100 nucleotides. This review provides insight into the resonance assignment methods that are the first and crucial step of all NMR studies, into the determination of base-pair geometry, into the examination of local and global RNA conformation, and into the detection of interaction sites of RNA. Examples of NMR investigations of RNA are given by using several different RNA molecules to illustrate the information content obtainable by NMR spectroscopy and the applicability of NMR techniques to a wide range of biologically interesting RNA molecules. PMID- 14523912 TI - Nucleic acid aptamers as tools and drugs: recent developments. AB - Nucleic acid aptamers are molecules that bind to their ligands with high affinity and specificity. Unlike other functional nucleic acids such as antisense oligonucleotides, ribozymes, or siRNAs, aptamers almost never exert their effects on the genetic level. They manipulate their target molecules such as gene products or epitopes directly and site specifically, leaving nontargeted protein functions intact. In a similar way to antibodies, aptamers bind to many different kinds of target molecules with high specificity and can be made to order, but as a result of their different biochemical nature and size they can also be used complementary to antibodies. In some cases, aptamers might be more suitable or more specific than antibody approaches or small molecules, both as scientific and biotechnological tools and as therapeutic agents. Recent examples of characterization of aptamers as tools for scientific research to study regulatory circuits, as tools in diagnostic or biosensor development, and as therapeutic agents are discussed. PMID- 14523913 TI - Targeting the HIV trans-activation responsive region--approaches towards RNA binding drugs. PMID- 14523914 TI - Spiegelmers: biostable aptamers. PMID- 14523915 TI - On secondary structure rearrangements and equilibria of small RNAs. PMID- 14523916 TI - Engineered ribozymes as molecular tools for site-specific alteration of RNA sequence. PMID- 14523917 TI - Targeting RNA with small molecules. PMID- 14523918 TI - Structural insight into functional aspects of ribosomal RNA targeting. PMID- 14523919 TI - RNA as a drug target: the case of aminoglycosides. PMID- 14523920 TI - Genetic control by metabolite-binding riboswitches. PMID- 14523921 TI - RNA-interference-based silencing of mammalian gene expression. PMID- 14523922 TI - Evaluation of bacterial RNase P RNA as a drug target. AB - RNA has gained increasing importance as a therapeutic target. However, so far mRNAs rather than stable cellular RNAs have been considered in such studies. In bacteria, the tRNA-processing enzyme RNase P has a catalytic RNA subunit. Fundamental differences in structure and function between bacterial and eukaryotic RNase P, and its indispensability for cell viability make the bacterial enzyme an attractive drug target candidate. Herein we describe two approaches utilized to evaluate whether the catalytic RNA subunit of bacterial RNase P is amenable to inactivation by antisense-based strategies. In the first approach, we rationally designed RNA hairpin oligonucleotides targeted at the tRNA 3'-CCA binding site (P15 loop region) of bacterial RNase P RNA by attempting to include principles derived from the natural CopA-CopT antisense system. Substantial inactivation of RNase P RNA was observed for Type A RNase P RNA (such as that in Escherichia coli) but not for Type B (as in Mycoplasma hyopneumoniae). Moreover, only an RNA oligonucleotide (Eco 3') complementary to the CCA binding site and its 3' flanking sequences was shown to be an efficient inhibitor. Mutation of Eco 3' and analysis of other natural RNase P RNAs with sequence deviations in the P15 loop region showed that inhibition is due to interaction of Eco 3' with this region and occurs in a highly sequence-specific manner. A DNA version of Eco 3' was a less potent inhibitor. The potential of Eco 3' to form an initial kissing complex with the P15 loop did not prove advantageous. In a second approach, we tested a set of oligonucleotides against E. coli RNase P RNA which were designed by algorithms developed for the selection of suitable mRNA targets. This approach identified the P10/11-J11/12 region of bacterial RNase P RNA as another accessible region. In conclusion, both the P15 loop and P10/11-J11/12 regions of Type A RNase P RNAs seem to be promising antisense target sites since they are easily accessible and sufficiently interspersed with nonhelical sequence elements, and oligonucleotide binding directly interferes with substrate docking to these two regions. PMID- 14523923 TI - Antisense inhibition of Escherichia coli RNase P RNA: mechanistic aspects. AB - The ribonucleoprotein enzyme RNase P catalyzes endonucleolytic 5'-maturation of tRNA primary transcripts in all domains of life. The indispensability of RNase P for bacterial cell growth and the large differences in structure and function between bacterial and eukaryotic RNase P enzymes comply with the basic requirements for a bacterial enzyme to be suitable as a potential novel drug target. We have identified RNA oligonucleotides that start to show an inhibitory effect on bacterial RNase P RNAs of the structural type A (for example, the Escherichia coli or Klebsiella pneumoniae enzymes) at subnanomolar concentrations in our in vitro precursor tRNA (ptRNA) processing assay. These oligonucleotides are directed against the so-called P15 loop region of RNase P RNA known to interact with the 3'-CCA portion of ptRNA substrates. Lead probing experiments demonstrate that a complementary RNA or DNA 14-mer fully invades the P15 loop region and thereby disrupts local structure in the catalytic core of RNase P RNA. Binding of the RNA 14-mer is essentially irreversible because of a very low dissociation rate. The association rate of this oligonucleotide is on the order of 10(4) M(-1) s(-1) and is thus comparable to those of many other artificial antisense oligonucleotides. The remarkable inhibition efficacy is attributable to the dual effect of direct interference with substrate binding to the RNase P RNA active site and induction of misfolding of the catalytic core of RNase P RNA. Based on our findings, the P15 loop region of bacterial RNase P RNAs of the structural type A can be considered the "Achilles' heel" of the ribozyme and therefore represents a promising target for combatting multiresistant bacterial pathogens. PMID- 14523924 TI - Structural investigation of a high-affinity MnII binding site in the hammerhead ribozyme by EPR spectroscopy and DFT calculations. Effects of neomycin B on metal ion binding. AB - Electron paramagnetic resonance spectroscopy and density functional theory methods were used to study the structure of a single, high-affinity Mn(II) binding site in the hammerhead ribozyme. This binding site exhibits a dissociation constant Ke of 4.4 microM in buffer solutions containing 1 M NaCl, as shown by titrations monitored by continuous wave (cw) EPR. A combination of electron spin echo envelope modulation (ESEEM) and hyperfine sublevel correlation (HYSCORE) experiments revealed that the paramagnetic manganese(II) ion in this binding site is coupled to a single nitrogen atom with a quadrupole coupling constant kappa of 0.7 MHz, an asymmetry parameter eta of 0.4, and an isotropic hyperfine coupling constant of Aiso(14N)=2.3 MHz. All three EPR parameters are sensitive to the arrangement of the Mn(II) ligand sphere and can therefore be used to determine the structure of the binding site. A possible location for this binding site may be at the G10.1, A9 site found to be occupied by Mn(II) in crystals (MacKay et al., Nature 1994, 372, 68 and Scott et al., Science 1996, 274, 2065). To determine whether the structure of the binding site is the same in frozen solution, we performed DFT calculations for the EPR parameters, based on the structure of the Mn(II) site in the crystal. Computations with the BHPW91 density function in combination with a 9s7p4d basis set for the manganese(II) center and the Iglo-II basis set for all other atoms yielded values of kappa(14N)=+0.80 MHz, eta=0.324, and Aiso(14N)=+2.7 MHz, in excellent agreement with the experimentally obtained EPR parameters, which suggests that the binding site found in the crystal and in frozen solution are the same. In addition, we demonstrated by EPR that Mn(II) is released from this site upon binding of the aminoglycoside antibiotic neomycin B (Kd=1.2 microM) to the hammerhead ribozyme. Neomycin B has previously been shown to inhibit the catalytic activity of this ribozyme (Uhlenbeck et al., Biochemistry 1995, 34, 11 186). PMID- 14523925 TI - Flexible adaptations in the structure of the tRNA-modifying enzyme tRNA-guanine transglycosylase and their implications for substrate selectivity, reaction mechanism and structure-based drug design. AB - The enzyme tRNA-guanine transglycosylase (TGT, EC 2.4.2.29) catalyses a base exchange reaction that leads to anticodon modifications of certain tRNAs. The TGT enzymes of the eubacteria Zymomonas mobilis (Z. mobilis TGT) and Escherichia coli (E. coli TGT) show a different behaviour in the presence of competitive inhibitors. The active sites of both enzymes are identical apart from a single conservative amino acid exchange, namely Tyr106 of Z. mobilis TGT is replaced by a Phe in E. coli TGT. Although Tyr106 is, in contrast to Phe106, hydrogen bonded in the ligand-free structure, we can show by a mutational study of TGT(Y106F) that this is not the reason for the different responses upon competition. The TGT enzymes of various species differ in their substrate selectivity. Depending on the applied pH conditions and/or induced by ligand binding, a peptide-bond flip modulates the recognition properties of the substrate binding site, which changes between donor and acceptor functionality. Furthermore interstitial water molecules play an important role in these adaptations of the pocket. The flip of the peptide bond is further stabilised by a glutamate residue that operates as general acid/base. An active-site aspartate residue, presumed to operate as a nucleophile through covalent bonding during the base-exchange reaction, shows different conformations depending on the nature of the bound ligand. The induced fit adaptations observed in the various TGT complex structures by multiple crystal-structure analyses are in agreement with the functional properties of the enzyme. In consequence, full understanding of this plasticity can be exploited for drug design. PMID- 14523926 TI - The molecular basis for A-site mutations conferring aminoglycoside resistance: relationship between ribosomal susceptibility and X-ray crystal structures. AB - Aminoglycoside antibiotics target the 16S ribosomal RNA (rRNA) bacterial A site and induce misreading of the genetic code. Point mutations of the ribosomal A site may confer resistance to aminoglycoside antibiotics. The influence of bacterial mutations (introduced by site-directed mutagenesis) on ribosomal drug susceptibility was investigated in vivo by determination of minimal inhibitory concentrations. To determine the origin of the various resistance phenotypes at a molecular level, the in vivo results were compared with the previously published crystal structures of paromomycin, tobramycin, and geneticin bound to oligonucleotides containing the minimal A site. Two regions appear crucial for binding in the A site: the single adenine residue at position 1408 and the non Watson-Crick U1406.U1495 pair. The effects of mutations at those positions are modulated by the nature of the substituent at position 6' (either hydroxy or ammonium group) on ring I, by the number of positive charges on the antibiotic, and by the linkage between rings I and III (either 4,5 or 4,6). In particular, the analysis demonstrates: 1) that the C1409-G1491 to A1409-U1491 polymorphism (observed in 15 % of bacteria) is not associated with resistance, which indicates that it does not affect the stacking of ring I on residue 1491, 2) that the high level resistance to 6'-NH3+ aminoglycosides exhibited by the A1408G mutation most probably results from the inability of ring I forming a pseudo base pair with G1408, which prevents its insertion inside the A site helix, and 3) that mutations of the uracil residues forming the U1406.U1495 pair either to cytosine or to adenine residues mostly confer low to moderate levels of drug resistance, whereas the U1406C/U1495A double mutation confers high-level resistance (except for neomycin), which suggests that aminoglycoside binding to the wild-type A site and its functional consequences strongly depend on a particular geometry of the U1406.U1495 pair. The relationships between the resistance phenotypes observed in vivo and the interactions described at the molecular level define the biological importance of the different structural interactions observed by X-ray crystallography studies. PMID- 14523927 TI - Stereoselective synthesis using immobilized Diels-Alderase ribozymes. AB - Development of artificial ribozymes by in vitro selection has so far, mostly been addressed from the viewpoint of fundamental research. However, such ribozymes also have high potential as selective catalysts in practical syntheses. Immobilization of an active and selective ribozyme is an important step towards this end. A 49-nucleotide RNA molecule that was previously found to stereoselectively catalyze Diels-Alder reactions between various anthracene dienes and maleimide dienophiles was quantitatively immobilized on an agarose matrix by periodate oxidation of the 3'-terminal ribose and coupling to a hydrazide moiety. Typical loadings were 45 pmol microL(-1) gel. The specific activity was comparable to that of soluble ribozyme, and high enantioselectivities were obtained in catalyzed cycloadditions. The catalytic matrix was found to be stable and could be regenerated about 40 times with only minimal reduction of catalytic activity. Like the soluble ribozyme, the immobilized catalyst stereoselectively converts various diene and dienophile substrates. By using either natural D-RNA or enantiomeric L-RNA, both product enantiomers were made synthetically accessible with similar selectivities. PMID- 14523928 TI - Identification of peptide ligands for target RNA structures derived from the HIV 1 packaging signal psi by screening phage-displayed peptide libraries. PMID- 14523932 TI - Cellular transport strategies for targeting epitopes, drugs and reporter molecules. Proceedings of a workshop. 6-9 March 2003, Budapest, Hungary. PMID- 14523933 TI - A brief introduction to cell-penetrating peptides. AB - Cell membranes act as protective walls to exclude most molecules that are not actively imported by living cells. This is an efficient way for a cell to prevent uncontrolled influx or efflux of solutes, which otherwise would be harmful to it. Only compounds within a narrow range of molecular size, polarity and net charge are able to diffuse effectively through cell membranes. In order to overcome this barrier for effective delivery of membrane-impermeable molecules, several chemical and physical methods have been developed. These methods, e.g. electroporation, and more recent methods as cationic lipids/liposomes, have been shown to be effective for delivering hydrophobic macromolecules. The drawbacks of these harsh methods are, primarily, the unwanted cellular effects exerted by them, and, secondly, their limitation to in vitro applications. The last decade's discovery of cell-penetrating peptides translocating themselves across cell membranes of various cell lines, along with a cargo 100-fold their own size, via a seemingly energy-independent process, opens up the possibility for efficient delivery of DNA, antisense peptide nucleic acids, oligonucleotides, proteins and small molecules into cells both in vitro and in vivo. PMID- 14523934 TI - Lipid vector for the delivery of peptides towards intracellular pharmacological targets. AB - The ability of single-chain lipopeptides to gain access to cellular compartments other than those related to degradation/recycling was first deduced from their capacity to deliver peptide antigens into MHC-class I loading mechanisms. The ability of lipopeptides to escape complete endosome degradation was further illustrated by the selective inhibition of different protein kinase C isoenzymes and, more recently, the presentation of agonistic activity towards the interferon gamma receptor. Taken together, several independent results indicate that modification of a peptide by a single lipid chain confers upon it intracellular trafficking properties that can be used to deliver functional cargo peptides into living cells; the endoplasmic reticulum, cytosolic protease activity, sites of kinase activity, or even the signalling pathway associated with cytokine stimulation, all appear accessible to peptide modified by a single lipidic moiety. In this context, the interferon gamma receptor can be considered as a very discriminative pharmacological model, useful for the comparative evaluation of the cellular delivery of lipopeptides, as it allows the unambiguous tracking of their intact delivery into a wide range of cellular compartments. This model is now being used to probe the influence of the nature of the lipid moiety on the trafficking properties of lipopeptides. PMID- 14523935 TI - Synthesis and application of functional peptides as cell nucleus-directed molecules in the treatment of malignant diseases. AB - The unique functions of biomolecules, including transport across biological membranes (e.g. the cell membrane, the nuclear envelope), modulation of protein function, gene transcription, reconstitution of the malignant transformation, and viral, bacterial and fungal activities underlie a high pharmaceutical potential. The development of combinatorial functional peptide modules in this important area has been slow, in contrast to the rapid development in the synthesis of small biopolymers. The conjugation of a short transmembrane transport peptide module with a cell nucleus address peptide module and with any substance is attractive for preparation of BioShuttle-based peptides because of the well established automated synthesis of peptides. Variation of the different functional modules for drug targeting and the choice of substances can be combined to create novel bioconjugates with unique properties. This article provides an overview of previous work on the BioShuttle technology and outlines the promising use of this approach in combinatorial peptide synthesis and drug discovery. PMID- 14523936 TI - Radiolabeled alpha-melanocyte-stimulating hormone analogs for receptor-mediated targeting of melanoma: from tritium to indium. AB - Following the first synthesis of tritiated alpha-melanocyte-stimulating hormone (alpha-MSH, alpha-melanotropin) in 1974 by Medzihradszky et al., several alpha MSH analogs were designed containing between 2 and 12 tritium atoms, the latter of which displayed a specific radioactivity of 12.21 GBq/micromol (330 Ci/mmol). Similarly, radioiodinated alpha-MSH analogs of high purity, full biological activity and a specific radioactivity of approximately 140 GBq/micromol were obtained. Although tritiated and radioiodinated alpha-MSH became indispensable tools as tracer molecules for numerous in vitro and in vivo studies, above all for receptor identification and characterization as well as for structure activity studies, they did not fulfill the criteria required for therapeutic in vivo targeting of metastatic melanoma. Therefore, we recently developed alpha-MSH analogs containing the universal metal chelator 1,4,7,10-tetraazacyclododecane 1,4,7,10-tetraacetic acid (DOTA) in different positions of the molecule. As DOTA can equally well incorporate diagnostic (e.g. (111)In, (67,68)Ga) and therapeutic (e.g. (90)Y, (67)Cu) radionuclides, DOTA-MSH compounds may serve for both melanoma scintigraphy and therapy. The analog DOTA-[betaAla(3), Nle(4), Asp(5), D Phe(7), Lys(10)]-alpha-MSH(3-10) (DOTA-MSH(OCT)), which contains the metal chelator at its N-terminal end, displayed good in vitro MC1R affinity (IC(50) 9.21 nm). In vivo, [(111)In]DOTA-MSH(OCT) exhibited a favorable biodistribution profile after injection in B16-F1 tumor-bearing mice. The radiopeptide was rapidly cleared from blood through the kidneys and, most importantly, accumulated preferentially in the melanoma lesions. Lung and liver melanoma metastases could be clearly imaged on tissue section autoradiographs 4 h after injection of [(111)In]DOTA-MSH(OCT). A comparative study of [(111)In]DOTA-MSH(OCT) with [(111)In]DOTA-[Nle(4), D-Phe(7)]-alpha-MSH ([(111)In]DOTA-NDP-MSH) demonstrated the superiority of the DOTA-MSH(OCT) peptide, particularly with respect to the amount of radioactivity taken up by non-malignant organs, including bone, the most radiosensitive tissue. These results demonstrate that [(111)In]DOTA-MSH(OCT) specifically targets melanoma metastases and represents a lead compound for the development of therapeutic DOTA-MSH analogs. PMID- 14523937 TI - Molecular tools for targeted imaging and therapy of cancer. AB - Here we review an approach to the design and production of antibody/ligand pairs for use in cell targeting procedures, to achieve functional affinity far greater than avidin/biotin. Using fundamental chemical principles, we have developed antibody/ligand pairs that retain the binding specificity of the antibody, but do not dissociate. By eliminating the dissociation of the ligand from the antibody, we have made the affinity functionally infinite. This methodology is applicable to other biological binding pairs. PMID- 14523938 TI - Membrane permeability commonly shared among arginine-rich peptides. AB - Delivery of proteins and other macromolecules using membrane-permeable carrier peptides is a recently developed novel technology, which enables us to modulate cellular functions for biological studies with therapeutic potential. One of the most often used carrier peptides is the arginine-rich basic peptide derived from HIV-1 Tat protein [HIV-1 Tat (48-60)]. Using this peptide, efficient intracellular delivery of molecules including proteins, oligonucleic acids and liposomes has been achieved. We have demonstrated that these features were commonly shared among many arginine-rich peptides such as HIV-1 Rev (34-50) and octaarginine. Not only the linear peptides but also branched-chain peptides showed efficient internalization with an optimum number of arginines (approximately eight residues). The structural and mechanistic features of the translocation of these membrane-permeable arginine-rich peptides are reviewed. PMID- 14523939 TI - Cellular uptake [correction of utake] of the Tat peptide: an endocytosis mechanism following ionic interactions. AB - The cellular delivery of various biological compounds has recently been improved by conjugating them to short peptides known as protein transduction domains or cell penetrating peptides. These peptides include Tat, Antennapedia and arginine rich peptides. The common feature of these peptides is their highly cationic nature. Up to now, the cellular uptake of about 50 different peptides and proteins coupled to Tat or Antennapedia peptides has been reported. The ability to deliver molecules into cells is not limited to peptide moieties, since oligonucleotides, peptide nucleic acids or other low molecular weight entities have been successfully internalized. Moreover, most of these examples have been accompanied by the expected biological response. More surprisingly, the uptake of large structures such as liposomes, phages, nanoparticles or adenoviruses has also been documented. Indeed the mechanism by which these very different entities could enter cells following a putative common pathway appeared more and more intriguing after each new reported example of cellular uptake mediated by these peptides. After a long period of uncertainty regarding the mechanism of entry, data from several groups now argue for an energy-dependent process of entry. The entry of most of these molecules is likely to be inhibited by low temperature incubation or in the presence of various drugs applied to inhibit the energy dependent pathway of cell entry. Moreover, the binding of the highly cationic Tat peptide to various anionic membrane components probably initiates the first step of the cell internalization process. PMID- 14523940 TI - Membrane translocation of penetratin and its derivatives in different cell lines. AB - The third helix of the homeodomain of the Antennapedia homeoprotein can translocate through the cell membrane into the nucleus and can be used as an intracellular vehicle for the delivery of oligopeptides and oligonucleotides. A 16-amino acid-long peptide fragment, called penetratin, is internalized by the cells in a specific, non-receptor-mediated manner. For a better understanding of the mechanism of the transfer, penetratin and two analogs were synthesized:The conformation of penetratin peptides 1-3 was examined in both extracellular matrix mimetic and membrane-mimetic environments. (1)H-NMR and CD spectroscopic measurements were performed in mixtures of TFE/water with different ratios. Peptides 1-3 were labeled by reacting their N-terminal free amino group with fluorescein isothiocyanate (FITC). Membrane translocation of the labelled peptides was studied with cell cultures [WEHI 164 murine fibrosarcoma cells (WC/1); chicken fibroblast cells (CEC-32); chicken monocytic cells (HD-11); human fibroblast (SV 80) and human monocytic cells (MonoMac-6)]. Confocal laser scanning microscopy and flow cytometry assay were used to study membrane translocation. Amphiphilicity was calculated for each peptide. In our experiments all the penetratin peptides penetrated into the cells. Helical conformation and membrane translocation ability showed little correlation: substitution of the two Trp with Phe increased the stability of helical conformation but decreased membrane translocation activity. The results of fluorescence microscopy and flow cytometry show that penetratin can be translocated into the cells by two mechanisms: endocytosis and direct transport through the cell membrane. PMID- 14523941 TI - Novel daunorubicin-carrier peptide conjugates derived from human calcitonin segments. AB - Severe and often therapy-limiting side effects are a major obstacle in cancer chemotherapy. New delivery concepts reducing systemic side effects are needed in order to optimize anticancer therapies. Several approaches have been followed, most of them concentrating on macromolecular carriers like liposomes, monoclonal antibodies, serum proteins or polyethylene glycol. We present here a novel type of anthracycline conjugate, using a small carrier peptide derived from the peptide hormone human calcitonin (hCT). The carrier peptide hCT(9-32) has so far been shown to be capable of transporting fluorophores or proteins across cellular membranes. Two different carrier peptide-daunorubicin conjugates were prepared, one with an acid-stable amide bond, the second with an acid-labile hydrazone bond. In vitro studies with daunorubicin linked to the carrier peptide via an acid-labile hydrazone bond demonstrated comparable cytotoxicity to daunorubicin in various daunorubicin sensitive cell lines (neuroblastoma cell lines SK-N-MC and SMS-KAN; HEK 293 T cells). In addition, fluorescence microscopy provided further insight into the mechanism of uptake of the carrier peptide hCT(9-32), indicating that endosomal compartments with reduced pH are involved in the intracellular release of daunorubicin. PMID- 14523943 TI - Targeting dendritic cells for priming cellular immune responses. AB - The cardinal role of dendritic cells (DC) in priming adaptive immunity and in orchestrating immune responses against all classes of pathogens and also against tumors is well established. Their unique potential both to maintain self tolerance and to initiate protective immune responses against foreign and/or dangerous structures is based on the functional diversity and flexibility of these cells. Tissue DC lining antigenic portals such as mucosal surfaces and the skin are specialized to take up a wide array of compounds including proteins, lipids, carbohydrates, glycoproteins, glycolipids and oligonucleotides, particles carrying such structures and apoptotic or necrotic cells. This process is facilitated by specialized receptors with high endocytic capacity, which provides potential targets for delivering designed molecules. The best route for targeting B- and/or T cell epitopes, however, is still the subject of intense investigation. Immature DC, which reside in various tissues, can be activated by pathogens, stress and inflammation or modified metabolic products, which induce mobilization of cells to draining lymph nodes where they act as highly potent professional antigen presenting cells. This is brought about by the ability to present their accumulated intracellular content for both CD4+ helper (Th) and CD8+ cytotoxic/cytolytic T lymphocytes (Tc/CTL). Engulfed proteins are processed intracellularly and their peptide fragments are transported to the cell surface in the context of major histocompatibility complex encoded class I and II molecules for presentation to Th cells and CTLs, respectively. The T cell priming capacity of DC, however, depends not only on antigen presentation but also on other features of DC. Human monocyte-derived DC provide an excellent tool to study the internalizing, antigen-presenting and T cell-activating functions of DC at their immature and activated differentiation states. These biological activities of DC, however, are highly dependent on their migratory potential from the peripheral non-lymphoid tissues to the lymph nodes, on the expression of adhesion molecules, which support the interaction of DC with T lymphocytes, and the cytokines secreted by DC, which polarize immune responses to Th1-mediated cellular or Th2-mediated antibody responses. These results altogether demonstrate that monocyte-derived DC are useful candidates for in vitro or in vivo targeting of antigens to induce efficient adaptive immune responses against pathogens and also against tumors. PMID- 14523942 TI - Drug targeting by macromolecules without recognition unit? AB - his review will summarize available information on the ability of macromolecular conjugates containing no specific recognition motifs to deliver anthracyclines (daunomycin, adriamycin) or methotrexate to target cells such as tumour cells or macrophages. Conjugates with natural (proteins, DNA, carbohydrates) and synthetic macromolecules (linear and branched chain poly-alpha-amino acids, non biodegradable DIVEMA, HPMA etc.) will be reviewed. Experimental data from several laboratories indicate that these conjugates are taken up by cells mainly by fluid phase or adsorptive endocytosis. It is believed that these processes do not involve 'specific receptors'. Two examples of methotrexate and daunomycin conjugates will be discussed to show the effect of the chemical structure of branched chain polypeptides on the uptake and antitumour or antiparasitic (Leishmania donovani infection) efficacy of conjugates. PMID- 14523944 TI - Targeting with scFv: immune modulation by complement receptor specific constructs. AB - The fate of a microbe in the host is determined by various molecules of the innate immune system, which recognize the microbe and enhance its interaction with antigen presenting cells. This 'natural targeting' phenomenon, however, does not function when antigens with limited immunogenicity enter the host. Peptide vaccines, for instance, require adjuvants to induce immune responses. As a surrogate for the natural targeting mechanisms, antibodies against selected receptors of antigen presenting cells, conjugated with the peptides, could be used as targeting devices. Here we review various antibody-mediated antigen targeting strategies, paying special attention to complement receptor-mediated targeting. We also describe and summarize our method of single-chain antibody mediated targeting of viral epitopes to complement receptor type two and discuss the perspectives of single-chain antibody-mediated antigen targeting. PMID- 14523945 TI - Recombinant antibodies with MHC-restricted, peptide-specific, T-cell receptor like specificity: new tools to study antigen presentation and TCR-peptide-MHC interactions. AB - The advent in recent years of the application of tetrameric arrays of class I peptide-MHC complexes now enables us to detect and study rare populations of antigen-specific CD8+ T cells. However, available methods cannot visualize or determine the number and distribution of these TCR ligands on individual cells or detect antigen-presenting cells (APCs) in tissues. Here we describe a new approach that enables study of human class I peptide-MHC ligand-presentation as well as TCR-peptide-MHC interactions. Such studies are facilitated by applying novel tools in the form of peptide-specific, HLA-A2-restricted human recombinant antibodies directed toward a large variety of tumor-associated as well as viral T cell epitope peptides. Using a large human antibody phage display library, a large panel of recombinant antibodies that are specific for a particular peptide MHC class I complex in a peptide-dependent, MHC-restricted manner was isolated. These antibodies were used to directly visualize the specific MHC-peptide complex on tumor cells, antigen-presenting cells or virus-infected cells by flow cytometry. They enabled direct quantitation of the number of MHC-peptide complexes as well as in situ detection of the complex on the surface of APCs after naturally occurring active intracellular processing of the cognate antigen. These studies will enable also the development of a new class of targeting molecules to deliver drugs or toxins to tumor or virus-infected cells. Thus, we demonstrate our ability to transform the unique fine specificity but low intrinsic affinity of TCRs into high-affinity soluble antibody molecules endowed with a TCR-like specificity toward human tumor or viral epitopes. These molecules may prove to be crucial useful tools for studying MHC class I antigen presentation in health and disease as well as for therapeutic purposes in cancer, infectious diseases and autoimmune disorders. PMID- 14523946 TI - The molecular basis of celiac disease. AB - Celiac disease is caused by inflammatory, gluten specific T cell responses in the small intestine. Invariably such responses are HLA-DQ2 or HLA-DQ8 restricted, providing an explanation for the strong association between celiac disease and these HLA-class II alleles. It is now clear that some native gluten sequences can bind to HLA-DQ2/8 and induce T cell responses. In addition, modification of gluten peptides by the enzyme tissue transglutaminase results in high affinity HLA-DQ2/8 binding peptides that can induce T cell responses. Thus, gluten molecules contain a large number of immunogenic peptides and this is likely to play an important role in the breaking of oral tolerance to gluten. PMID- 14523947 TI - Destination 'lysosome': a target organelle for tumour cell killing? AB - Lysosomes and lysosome-related organelles constitute a system of acid compartments that interconnect the inside of the cell with the extracellular environment via endocytosis, phagocytosis and exocytosis. In recent decades it has been recognized that lysosomes are not just wastebaskets for disposal of unused cellular constituents, but that they are involved in several cellular processes such as post-translational maturation of proteins, degradation of receptors and extracellular release of active enzymes. By complementing the autophagic process, lysosomes actively contribute to the maintenance of cellular homeostasis. Proteolysis by lysosomal cathepsins has been shown to mediate the death signal of cytotoxic drugs and cytokines, as well as the activation of pro survival factors. Secreted lysosomal cathepsins have been shown to degrade protein components of the extracellular matrix, thus contributing actively to its re-modelling in physiological and pathological processes. The malfunction of lysosomes can, therefore, impact on cell behaviour and fate. Here we review the role of lysosomal hydrolases in several aspects of the malignant phenotype including loss of cell growth control, altered regulation of cell death, acquisition of chemoresistance and of metastatic potential. Based on these observations, the lysosome is proposed as a potential target organelle for the chemotherapy of tumours. We will also present some recent data concerning the technologies for delivering chemotherapeutic drugs to the endosomal-lysosomal compartment and the strategies to improve their efficacy. PMID- 14523948 TI - Sources of plasma glucose by automated Bayesian analysis of 2H NMR spectra. AB - Sources of blood glucose can be determined after oral ingestion of (2)H(2)O followed by isolation of plasma glucose and measurement of the relative (2)H enrichments in select positions within the glucose molecule. Typically, (2)H enrichments are obtained by mass spectrometry but (2)H NMR offers an alternative. Here it is demonstrated that the entire analysis may be automated by Bayesian analysis of a (2)H free induction decay signal of monoacetone glucose to obtain a direct readout of the relative contributions of glycogenolysis, glycerol, and phosphoenol pyruvate to plasma glucose production. Furthermore, Markov Chain Monte Carlo (MCMC) simulations of the posterior probability density provide uncertainties in all metabolic parameters from a single patient, thereby allowing comparisons in glucose metabolism from one individual to another. The combined MCMC Bayesian methodology is operationally simple and requires little intervention from the operator. PMID- 14523949 TI - Statistical model for diffusion attenuated MR signal. AB - A general statistical model that can describe a rather large number of experimental results related to the structure of the diffusion-attenuated MR signal in biological systems is introduced. The theoretical framework relies on a phenomenological model that introduces a distribution function for tissue apparent diffusion coefficients (ADC). It is shown that at least two parameters- the position of distribution maxima (ADC) and the distribution width (sigma)--are needed to describe the MR signal in most regions of a human brain. A substantial distribution width, on the order of 36% of the ADC, was found for practically all brain regions examined. This method of modeling the MR diffusion measurement allows determination of an intrinsic tissue-specific ADC for a given diffusion time independent of the strength of diffusion sensitizing gradients. The model accounts for the previously found biexponential behavior of the diffusion attenuated MR signal in CNS. PMID- 14523950 TI - Neurobehavioral effects among subjects exposed to high static and gradient magnetic fields from a 1.5 Tesla magnetic resonance imaging system--a case crossover pilot study. AB - The interactive use of magnetic resonance imaging (MRI) techniques is increasing in operating theaters. A study was performed on 17 male company volunteers to assess the neurobehavioral effects of exposure to magnetic fields from a 1.5 Tesla MRI system. The subjects' neurobehavioral performances on a neurobehavioral test battery were compared in four 1-hr sessions with and without exposure to magnetic fields, and with and without additional movements. Adverse effects were found for hand coordination (-4%, P < 0.05; Pursuit Aiming II) and near visual contrast sensitivity (-16% and -15%, P < 0.10; Vistech 6000). The results from the remaining tests were inconclusive due to a strong learning effect. No additional effect from gradient fields was detected. The results indicate that working near a 1.5 Tesla MRI system may lead to neurobehavioral effects. Further research is recommended, especially in members of operating teams using interactive MRI systems. PMID- 14523951 TI - Functional brain imaging using a blood oxygenation sensitive steady state. AB - Blood oxygenation level dependent (BOLD) functional MRI (fMRI) is an important method for functional neuroimaging that is sensitive to changes in blood oxygenation related to brain activation. While BOLD imaging has good spatial coverage and resolution relative to other neuroimaging methods (such as positron emission tomography (PET)), it has significant limitations relative to other MRI techniques, including poor spatial resolution, low signal levels, limited contrast, and image artifacts. These limitations derive from the coupling of BOLD functional contrast to sources of image degradation. This work presents an alternative method for fMRI that may over-come these limitations by establishing a blood oxygenation sensitive steady-state (BOSS) that inverts the signal from deoxygenated blood relative to the water signal. BOSS fMRI allows the imaging parameters to be optimized independently of the functional contrast, resulting in fewer image artifacts and higher signal-to-noise ratio (SNR). In addition, BOSS fMRI has greater functional contrast than BOLD. BOSS fMRI requires careful shimming and multiple acquisitions to obtain a precise alignment of the magnetization to the SSFP frequency response. PMID- 14523952 TI - 1H-localized broadband 13C NMR spectroscopy of the rat brain in vivo at 9.4 T. AB - Localized (13)C NMR spectra were obtained from the rat brain in vivo over a broad spectral range (15-100 ppm) with minimal chemical-shift displacement error (<10%) using semi-adiabatic distortionless enhancement by polarization transfer (DEPT) combined with (1)H localization. A new gradient dephasing scheme was employed to eliminate unwanted coherences generated by DEPT when using surface coils with highly inhomogeneous B(1) fields. Excellent sensitivity was evident from the simultaneous detection of natural abundance signals for N-acetylaspartate, myo inositol, and glutamate in the rat brain in vivo at 9.4 T. After infusion of (13)C-labeled glucose, up to 18 (13)C resonances were simultaneously measured in the rat brain, including glutamate C2, C3, C4, glutamine C2, C3, C4, aspartate C2, C3, glucose C1, C6, N-acetyl-aspartate C2, C3, C6, as well as GABA C2, lactate C3, and alanine C3. (13)C-(13)C multiplets corresponding to multiply labeled compounds were clearly observed, suggesting that extensive isotopomer analysis is possible in vivo. This unprecedented amount of information will be useful for metabolic modeling studies aimed at understanding brain energy metabolism and neurotransmission in the rodent brain. PMID- 14523953 TI - Scalar couplings as pH probes in compartmentalized biological systems: 31P NMR of phosphite. AB - The use of scalar couplings in nuclear magnetic resonance (NMR) spectra was investigated as a possible tool for the measurement of pH in different compartments of biological systems. The proposed method is attractive because no internal reference is required, unlike more widely used chemical shift titrations. The phosphite anion is shown to be ideal for the measurement of pH in the physiological range. In isotonic solution, the divalent anion PHO(3)(2-) has a one-bond (1)J(PH) of 568.1 Hz, increasing to 620.7 Hz for the monovalent anion PH(OH)O(2)(-), with a measured pK(a) of 6.19. The technique was applied to the measurement of pH in a suspension of human erythrocytes. The (31)P spectra of these species give well-resolved doublets for the intra- and extracellular spaces, and can be used to measure the pH difference across the cell membrane with errors on the order of about 0.01 pH units in the range of pH 5-7. Variations in erythrocyte pH due to metabolic changes are clearly observed. PMID- 14523954 TI - Independent component analysis for automated decomposition of in vivo magnetic resonance spectra. AB - Fully automated methods for analyzing MR spectra would be of great benefit for clinical diagnosis, in particular for the extraction of relevant information from large databases for subsequent pattern recognition analysis. Independent component analysis (ICA) provides a means of decomposing signals into their constituent components. This work investigates the use of ICA for automatically extracting features from in vivo MR spectra. After its limits are assessed on artificial data, the method is applied to a set of brain tumor spectra. ICA automatically, and in an unsupervised fashion, decomposes the signals into interpretable components. Moreover, the spectral decomposition achieved by the ICA leads to the separation of some tissue types, which confirms the biochemical relevance of the components. PMID- 14523955 TI - Reproducibility of 1H-MRS measurements in schizophrenic patients. AB - The reproducibility of single-voxel spectroscopic measures in a stable schizophrenic patient population is reported. Spectra acquired 1 week apart, in the left frontal white matter and left caudate nucleus of 12 patients, were analyzed using both frequency and time-domain techniques. No significant change in mean concentrations for N-acetylaspartate (NAA), choline (Cho), and creatine (Cre) were found between sessions. Variation from scan to scan was low for the left frontal white matter, with coefficients of variation (CV) < 5% for all three metabolites. NAA measures from the left caudate nucleus had similar reproducibility (CV = 5%), but Cho and Cre were poorer by comparison (CV = 16 18%). The two methods used for spectral analyses produced similar results, indicating that both techniques are appropriate. These results extend previous studies of healthy, normal subjects, and establish that it is possible to obtain reliable measures of brain metabolites in clinical populations. PMID- 14523956 TI - Impact of intravascular signal on quantitative measures of cerebral oxygen extraction and blood volume under normo- and hypercapnic conditions using an asymmetric spin echo approach. AB - An asymmetric spin echo (ASE) single shot echo planar imaging (EPI) sequence is proposed for obtaining quantitative estimates of R2', cerebral venous blood volume fraction (vCBV), and oxygen extraction fraction (OEF) noninvasively in normal volunteers. The impact of the presence of intravascular signal on the estimates of vCBV and OEF were examined in five subjects with different levels of flow attenuation. A significant reduction in the estimates of vCBV and a small increase in the measurements of OEF were observed in the presence of flow suppression gradients. In addition, mild hypercapnia was induced in normal subjects (n = 4). R2', vCBV, and OEF were measured under both normocapnia and experimentally induced hypercapnia. In agreement with the well-documented cerebral vascular responses to hypercapnia, estimates of R2' and OEF decrease, while measures of vCBV increase during hypercapnia. PMID- 14523957 TI - Magnetic resonance measurement of tetramethylammonium diffusion in rat brain: Comparison of magnetic resonance and ionophoresis in vivo diffusion measurements. AB - Magnetic resonance (MR) and ionophoresis are two experimental methods that provide measurements of molecular diffusion in living tissue. Typical experimental settings yield MR studies that are sensitive to mean molecular displacements of approximately 5 microm, and ionophoresis experiments to displacements of > or =100 microm. An assessment of the correspondence between the methods is hampered by the fact that no common probe molecule has been used. One of the most frequently utilized probe molecules in ionophoresis measurements is the tetramethylammonium (TMA) ion. In the current work the diffusion properties of TMA were studied in rat brain in vivo with localized (1)H MR spectroscopy (MRS). Standard treatment of the MR data yielded a 3.6-fold lower apparent diffusion coefficient (ADC) compared to ionophoresis. To explore the source of this discrepancy, a separate data processing scheme was applied to the MR data to monitor individual elapsed displacement-distance subpopulations of TMA molecules. This analysis revealed a dependence of the ADC estimation on a given subpopulation's elapsed displacement distance. The MR-derived ADC approached the ionophoresis-derived value as the elapsed displacement distance increased to 15 microm. These observations demonstrate that MR and ionophoresis studies provide complementary information, and that ADC estimates obtained from the two techniques are sensitive to different biophysical determinants. PMID- 14523958 TI - Characterization of continuously distributed cortical water diffusion rates with a stretched-exponential model. AB - Experience with diffusion-weighted imaging (DWI) shows that signal attenuation is consistent with a multicompartmental theory of water diffusion in the brain. The source of this so-called nonexponential behavior is a topic of debate, because the cerebral cortex contains considerable microscopic heterogeneity and is therefore difficult to model. To account for this heterogeneity and understand its implications for current models of diffusion, a stretched-exponential function was developed to describe diffusion-related signal decay as a continuous distribution of sources decaying at different rates, with no assumptions made about the number of participating sources. DWI experiments were performed using a spin-echo diffusion-weighted pulse sequence with b-values of 500-6500 s/mm(2) in six rats. Signal attenuation curves were fit to a stretched-exponential function, and 20% of the voxels were better fit to the stretched-exponential model than to a biexponential model, even though the latter model had one more adjustable parameter. Based on the calculated intravoxel heterogeneity measure, the cerebral cortex contains considerable heterogeneity in diffusion. The use of a distributed diffusion coefficient (DDC) is suggested to measure mean intravoxel diffusion rates in the presence of such heterogeneity. PMID- 14523959 TI - Effects of barrier-induced nuclear spin magnetization inhomogeneities on diffusion-attenuated MR signal. AB - The spatial distribution of the transverse nuclear spin magnetization, appearing in a single compartment with impermeable boundaries in a Stejskal-Tanner gradient pulse MR experiment, is analyzed in detail. At short diffusion times the presence of diffusion-restrictive barriers (membranes) reduces effective diffusivity near the membranes and leads to an inhomogeneous spin magnetization distribution (the edge-enhancement effect). In this case, the signal reveals a quasi-two compartment behavior and can be empirically modeled remarkably well by a biexponential function. The current results provide a framework for interpreting experimental MR data on various phenomena, including water diffusion in giant axons, metabolite diffusion in the brain, and hyperpolarized gas diffusion in lung airways. PMID- 14523960 TI - Relative indices of water diffusion anisotropy are equivalent in live and formalin-fixed mouse brains. AB - Formalin fixation of tissue is a common laboratory practice. A direct comparison of diffusion tensor imaging (DTI) parameters from mouse brains before (in vivo) and after (ex vivo) formalin fixation is reported herein. Five diffusion indices were examined in a cohort of seven mice: relative anisotropy (RA), directional correlation (DC), trace (Tr(D)), trace-normalized axial diffusivity (D(axially)), and radial diffusivity (D(radially)). Seven regions of interest (ROIs), including five in white matter and two in gray matter, were selected for examination. Consistent with previous findings, a significant decrease of Tr(D) was observed for all ROIs after fixation. However, water diffusion anisotropy, as defined by the indices RA, DC, D(axially), and D(radially), remained unchanged after fixation. Thus, fixation does not appear to alter diffusion anisotropy in the mouse brain. This finding supports the utility of diffusion anisotropy analysis of fixed tissue. The combination of DTI measurements and standard histology may shed light on the microstructural determinants of diffusion anisotropy in normal and disease states. PMID- 14523961 TI - Normative pediatric brain data for spatial normalization and segmentation differs from standard adult data. AB - Spatial normalization and morphological studies of pediatric brain imaging data based on adult reference data may not be appropriate due to the developmental differences between the two populations. In this study, we set out to create pediatric templates and a priori brain tissue data from a large collection of normal, healthy children to compare it to standard adult data available within a widely used imaging software solution (SPM99, WDOCN, London, UK). Employing four different processing strategies, we found considerable differences between our pediatric data and the adult data. We conclude that caution should be used when analyzing pediatric brain data using adult a priori information. To assess the effects of using pediatric a priori brain information, the data obtained in this study is available to the scientific community from our website (www.irc.cchmc.org). PMID- 14523962 TI - Comparison of dendrimer-based macromolecular contrast agents for dynamic micro magnetic resonance lymphangiography. AB - Few methods are currently available to visualize the entire lymphatic system. A method known as micro-magnetic resonance lymphangiography (MRL), which employs a dendrimer-based MRI contrast agent (PAMAM-G8) and a clinical-grade 1.5T MRI instrument, was recently developed for use in mice. In the present study, three dendrimer-based MRI contrast agents (PAMAM-G8, DAB-G5, and PAMAM-G4) with different pharmacokinetic characteristics were compared to determine the best reagent to visualize the lymphatic system under physiological or pathological conditions. In addition, two established MRI contrast agents (Gadomer-17 and Gd [DTPA]-dimeglumine (Magnevist)) were used as control agents. In experiments with mice, most of the deep lymphatic system was visualized by micro-MRL with all agents except Gd-[DTPA]-dimeglumine. PAMAM-G8 was best for visualizing lymphatic vessels, whereas DAB-G5 was better for visualizing lymph nodes. PAMAM-G4 was intermediate in character between PAMAM-G8 and DAB-G5, except in exhibiting a low background signal (especially in the liver). The lymphatic system was not clearly visualized with Gd-[DTPA]-dimeglumine; however, the lymph nodes were visualized with Gadomer-17, although not as well as with dendrimer-based agents. In conclusion, DAB-G5 and PAMAM-G4 can be used to identify lymph nodes and lymphatic vessels, respectively. Their rapid excretion makes these compounds potentially attractive for human use. PMID- 14523963 TI - Imaging the fate of implanted bone marrow stromal cells labeled with superparamagnetic nanoparticles. AB - Bone marrow stromal cells (MSCs) are pluripotent progenitor cells that have the capacity to migrate toward lesions and induce or facilitate site-dependent differentiation in response to environmental signals. In animals with a cortical photochemical lesion, the fate of rat MSCs colabeled with magnetic iron-oxide nanoparticles (Endorem) and bromodeoxyuridine (BrdU) was studied. MSCs were either grafted intracerebrally into the contralateral hemisphere of adult rat brain or injected intravenously. In vivo MRI was used to track their fate; Prussian blue staining and transmission electron microscopy (TEM) confirmed the presence of iron-oxide nanoparticles inside the cells. During the first week posttransplantation, the transplanted cells migrated to the lesion site and populated the border zone of the damaged cortical tissue. The implanted cells were visible on MR images as a hypointense area at the injection site and in the lesion. The hypointense signal persisted for more than 50 days. The presence of BrdU-positive and iron-containing cells was confirmed by subsequent histological staining. Three to 4 weeks after injection, <3% of MSCs around the lesion expressed the neuronal marker NeuN. Our study demonstrates that a commercially available contrast agent can be used as a marker for the long-term noninvasive MR tracking of implanted cells. PMID- 14523964 TI - Dynamic 3He imaging for quantification of regional lung ventilation parameters. AB - Dynamic ventilation imaging using laser-polarized (3)He has a promising potential for elucidating the physiology and physiopathology of the lungs. In this study, a methodological approach is proposed for the assessment and quantification of local ventilation parameters. High-temporal-resolution coronal ventilation image series were obtained with a projection-reconstruction (PR) sequence combined with the sliding-window technique. After image series were processed, parametric pixel by-pixel maps of the gas arrival time, filling time constant, inflation rate, and gas volume were generated. The acquisition technique and the signal processing procedure, which are referred to collectively as sliding pulmonary imaging for respiratory overview (SPIRO), were tested in vivo in healthy rat lungs using a contrast media injector for controlled (3)He flow and volume injection in the animal lungs. The same protocol was applied to broncho-constriction animal models using intravenous injection of methacholine solution. Inflation rate values measured in the lungs were found to decrease with increasing doses of injected methacholine solution. This study demonstrates that it is possible to obtain quantitative regional gas dynamic information using the SPIRO technique in a single polarized gas inspiration. PMID- 14523965 TI - Passive tracking exploiting local signal conservation: the white marker phenomenon. AB - This article presents a novel approach to passive tracking of paramagnetic markers during endovascular interventions, exploiting positive contrast of the markers to their background, so-called "white marker tracking." The positive contrast results from dephasing of the background signal with a slice gradient, while near the marker the signal is conserved because a dipole field induced by the marker compensates the dephasing gradient. Theoretical investigation shows that a local gradient induced by the local dipole field will nearly always cancel the dephasing gradient somewhere, regardless of marker composition, gradient strength, orientation, and acquisition parameters. The actual appearance of the white marker is determined by the marker strength, echo-time, slice thickness, and gradient strength, as shown both theoretically and experimentally. The novel concept is demonstrated by tracking experiments in a flow phantom and in pig models and is shown to allow reliable and robust depiction of paramagnetic markers with positive contrast and significant suppression of the background signal. PMID- 14523966 TI - Generalized reconstruction of phase contrast MRI: analysis and correction of the effect of gradient field distortions. AB - To characterize gradient field nonuniformity and its effect on velocity encoding in phase contrast (PC) MRI, a generalized model that describes this phenomenon and enables the accurate reconstruction of velocities is presented. In addition to considerable geometric distortions, inhomogeneous gradient fields can introduce deviations from the nominal gradient strength and orientation, and therefore spatially-dependent first gradient moments. Resulting errors in the measured phase shifts used for velocity encoding can therefore cause significant deviations in velocity quantification. The true magnitude and direction of the underlying velocities can be recovered from the phase difference images by a generalized PC velocity reconstruction, which requires the acquisition of full three-directional velocity information. The generalized reconstruction of velocities is applied using a matrix formalism that includes relative gradient field deviations derived from a theoretical model of local gradient field nonuniformity. In addition, an approximate solution for the correction of one directional velocity encoding is given. Depending on the spatial location of the velocity measurements, errors in velocity magnitude can be as high as 60%, while errors in the velocity encoding direction can be up to 45 degrees. Results of phantom measurements demonstrate that effects of gradient field nonuniformity on PC-MRI can be corrected with the proposed method. PMID- 14523967 TI - RINGLET motion correction for 3D MRI acquired with the elliptical centric view order. AB - A new rigid-body motion correction algorithm is described that is compatible with 3D image sets acquired with the elliptical centric (EC) view order. With this view order, an annular ring of k-space data is acquired in the ky-kz plane during any short time interval. Images for tracking motion can be reconstructed in the yz-plane from any ring of the acquisition data. In these tracking images, a point source (such as an external marker) shows a characteristic bull's-eye pattern that permits motion monitoring and correction. The true position of the point object is located at the center of the bull's-eye pattern. Cross correlation can be performed to automatically track the positions of markers reconstructed from adjacent rings of k-space. To increase the marker signal, the markers are encased in inductively coupled RF coils. Rigid-body motion in the yz-plane is calculated directly with the Euclidean group for rotation and translation, and corrected by rotating and applying phase shifts to any corrupted rings of data. In the current work we present a theoretical analysis of this method, as well as results of volunteer and controlled phantom experiments that demonstrate its initial feasibility. Although the EC view order has mainly been used for MR angiography (MRA), it can also be used for most 3D acquisitions. PMID- 14523968 TI - Importance of k-space trajectory in echo-planar myocardial tagging at rest and during dobutamine stress. AB - Hybrid fast gradient echo/echo-planar imaging (FGRE-EPI) can be used to increase temporal resolution, enhance tag contrast, and/or decrease scan time for breathhold myocardial tagging. However, off-resonance effects and motion can lead to local phase discontinuities in FGRE-EPI raw data when a conventional interleaved bottom-up k-space trajectory is used. These discontinuities can be particularly problematic for myocardial tagging, where the image energy is not only concentrated near the k-space origin, but is also concentrated in multiple spectral peaks centered throughout k-space. In this study, tag distortion artifacts in FGRE-EPI tagging due to off-resonance and velocity-induced phase discontinuities were characterized at rest and dobutamine stress, and the flyback and gradient moment smoothing (GMS) methods were shown to reduce these artifacts. For the specific parameters used in this study, flyback and GMS resulted in improved image quality at rest and stress, increased myocardium-tag contrast-to noise ratio (11.4 +/- 2.1 vs. 10.0 +/- 2.9, P < 0.01 at rest; 11.1 +/- 1.8 vs. 8.1 +/- 2.4, P < 0.01 at stress), and reduced full width at half maximum of the tag profile (3.6 vs. 3.8 pixels at rest; 4.0 vs. 5.1 pixels at stress) compared to the conventional trajectory. A limitation of the improved trajectory is a parameter-dependent decrease in data acquisition efficiency. For the specific imaging protocol used, the repetition time of the improved trajectory increased by 36% compared to the conventional trajectory. PMID- 14523969 TI - High-resolution diffusion-weighted 3D MRI, using diffusion-weighted driven equilibrium (DW-DE) and multishot segmented 3D-SSFP without navigator echoes. AB - In this work we report on the development of a novel technique for high resolution diffusion-weighted (DW) MRI based upon 3D steady-state free precession (3D-SSFP). First the 3D-SSFP acquisition was segmented (each segment consisting of a series of RF pulses and gradient-recalled echoes), and then DW-driven equilibrium (DE) was inserted between each segment. The in-plane imaging matrix was typically 256 x 192 or 256 x 160, which resulted in high-resolution DW images. The DW-DE segmented SSFP signal was contaminated by the non-DW magnetization, which recovered and contributed signal during the readout train (T(1) contamination). Center-out slice encoding was used to place the greatest diffusion weighting at the center of k-space. A numerical simulation and supporting experiments were performed to evaluate the relationship of the transverse magnetization to imaging parameters, such as the b-value, echo-train length (ETL), echo-train (group) repetition time (TR(g)), and RF excitation TR (Delta t). Both the numerical simulation and the experiments suggested that the effect of T(1) contamination would be reduced with a longer TR(g), smaller b value, shorter ETL, and center-out slice phase encoding. Phase errors caused by microscopic motions during the diffusion gradients were converted into amplitude errors by the tip-up pulse at the end of the diffusion-weighting segment. As a result, small bulk motions, such as CSF pulsation, did not cause motion-related ghosting artifacts, which would be typical in images from other multishot DWI techniques. This technique can be used for high-resolution DWI of nonbrain anatomies. PMID- 14523970 TI - A PRESTO-SENSE sequence with alternating partial-Fourier encoding for rapid susceptibility-weighted 3D MRI time series. AB - A 3D sequence for dynamic susceptibility imaging is proposed which combines echo shifting principles (such as PRESTO), sensitivity encoding (SENSE), and partial Fourier acquisition. The method uses a moderate SENSE factor of 2 and takes advantage of an alternating partial k-space acquisition in the "slow" phase encode direction allowing an iterative reconstruction using high-resolution phase estimates. Offering an isotropic spatial resolution of 4 x 4 x 4 mm(3), the novel sequence covers the whole brain including parts of the cerebellum in 0.5 sec. Its temporal signal stability is comparable to that of a full-Fourier, full-FOV EPI sequence having the same dynamic scan time but much less brain coverage. Initial functional MRI experiments showed consistent activation in the motor cortex with an average signal change slightly less than that of EPI. PMID- 14523971 TI - Single-shot magnetic field mapping embedded in echo-planar time-course imaging. AB - A technique for acquiring magnetic field maps simultaneously with gradient recalled echo-planar time-course data is described. This technique uses a trajectory in which the central part of k-space is collected twice. For a 64 x 64 image acquired with a 125-kHz bandwidth, a field map suitable for geometric correction can be collected simultaneously with the echo-planar time-course data in <70 ms. The field maps generated by this technique are registered with the magnitude images because they are calculated using the same data. They do not suffer from errors due to subject motion, or from different geometric distortions that can result from using different pulse sequences. In addition to correcting geometric distortions that resulted from dynamic magnetic field perturbations, this method was used to measure field shifts arising from respiration and jaw motion across five subjects. Values ranged from 0.035 to 0.165 parts per million (ppm). PMID- 14523972 TI - Cerebral blood volume measurements by T*2-weighted MRI and contrast infusion. AB - A reliable, accurate, and accessible method for measuring cerebral blood volume (CBV) has been developed based on T(*) (2)-weighted MRI and a 1-min infusion of gadolinium instead of a bolus. Computer simulations predict that this infusion CBV method will have a signal-to-noise ratio (SNR) 3-5 times greater than that obtained by area-under-the-curve (AUC) methods, with high accuracy over a wide range of arterial, tissue, and MRI conditions. In six healthy controls, the CBV was 1.87 +/- 0.44 in white matter (WM), 3.40 +/- 0.44 in deep gray matter (DGM), and 3.84 +/- 1.87 ml blood/100 g tissue in cortical GM (CGM). The mean GM/WM ratio was 1.94. In five patients with bilateral carotid disease, the corresponding values were 2.63 +/- 0.33, 4.72 +/- 0.33, and 5.27 +/- 2.40 ml blood/100 g tissue, all of which were significantly different from controls. AUC values were generally higher and failed to demonstrate differences between controls and patients. The infusion method shows great potential for providing reliable, accurate, and accessible CBV values with the ability to discriminate physiologic or pathological volume changes under a wide range of conditions. PMID- 14523974 TI - Application of continuous-wave EPR spectral-spatial image reconstruction techniques for in vivo oxymetry: comparison of projection reconstruction and constant-time modalities. AB - In this study we report the application of continuous-wave (CW) electron paramagnetic resonance (EPR) constant-time spectral spatial imaging (CTSSI) for in vivo oxymetry. 2D and 3D SSI studies of a phantom and live mice were carried out using projection reconstruction (PR) and constant-time (CT) modalities using a CW-EPR spectrometer/imager operating at 300 MHz frequency. Distortion of line shape, which is inherent in the PR method, was minimized by the CTSSI modality. It was also found that CTSSI offers improved noise reduction, restores a smoother line shape, and gives high convergence of estimated values. Spatial resolution was also improved by CTSSI, although fundamental spectral line-width broadening was observed. Although additional corrections are required for accurate estimations of spectral line width, CTSSI was able to demonstrate distinct differences in oxygen tension between a tumor and the normal legs of a C3H mouse. The PR method, on the other hand, was unable to make such a distinction unequivocally with the triarylmethyl spin probes. CTSSI promises to be a more suitable method for quantitative in vivo oxymetric studies using radiofrequency EPR imaging (EPRI). PMID- 14523973 TI - Effects of tracer arrival time on flow estimates in MR perfusion-weighted imaging. AB - A common technique for calculating cerebral blood flow (CBF) and mean transit time (MTT) is to track a bolus of contrast agent using perfusion-weighted MRI (PWI) and to deconvolve the change in concentration with an arterial input function (AIF) using singular value decomposition (SVD). This method has been shown to often overestimate the volume of tissue that infarcts and in cases of severe vasculopathy to produce CBF maps that are inconsistent with clinical presentation. This study examines the effects of tracer arrival time differences between tissue and a user-selected global AIF on flow estimates. CBF and MTT were calculated in both numerically simulated and clinically acquired PWI data where the AIF and tissue signals were shifted backward and forward in time with respect to one another. Results show that when the AIF leads the tissue, CBF is underestimated independent of extent of delay, but dependent on MTT. When the AIF lags the tissue, flow may be over- or underestimated depending on MTT and extent of timing differences. These conditions may occur in practice due to the application of a user-selected AIF that is not the "true AIF" and therefore caution must be taken in interpreting CBF and MTT estimates. PMID- 14523976 TI - High flip angle imaging of metallic stents: implications for MR angiography and intraluminal signal interpretation. AB - Intraluminal stent signal characterization by MRI is generally hampered by signal loss from the metallic stent material. This signal loss is related to magnetic susceptibility and RF shielding. Even when stent materials with low magnetic susceptibility are used, RF shielding can still be problematic. In this article we have shown that high flip angle imaging enables morphology assessment and tissue characterization in stents made of stainless steel 316L, NiTinol, and ABI alloy. PMID- 14523975 TI - Electrical conductivity imaging by magnetic resonance electrical impedance tomography (MREIT). AB - Magnetic resonance electrical impedance tomography (MREIT) is a recently developed imaging technique that combines MRI and electrical impedance tomography (EIT). In MREIT, cross-sectional electrical conductivity images are reconstructed from the internal magnetic field density data produced inside an electrically conducting subject when an electrical current is injected into the subject. In this work the results of an electrical conductivity imaging experiment are presented, along with some practical considerations regarding MREIT. The MREIT experiment was performed with a 0.3 Tesla MRI system on a phantom made of two compartments with different electrical conductivities. The current density inside the phantom was measured by the MR current density imaging (MRCDI) technique. The measured current density was then used for conductivity image reconstruction by the J-substitution algorithm. The conductivity phantom images obtained with an injection current of 28mA showed conductivity errors of about 25.5%. PMID- 14523977 TI - Imaging gene expression using oligonucleotides and peptide nucleic acids. AB - The development of methods for non-invasive, real-time imaging of gene expression would provide powerful tools for biomedical research and medical diagnostics. A broadly applicable strategy for achieving this goal is the use of complementary oligonucleotide probes for recognition of mRNA. The major challenge for molecular imaging is the development of specific and efficient transducers for signaling probe-target interaction. This review summarizes the strengths and limitations of reported molecular approaches for imaging of mRNA expression and discusses the challenges to development of in vivo methods. PMID- 14523978 TI - Molecular imaging: The latest generation of contrast agents and tissue characterization techniques. AB - Molecular Imaging technologies will have a profound impact on both basic research and clinical imaging in the near future. As the field covers many different specialties and scientific disciplines it is not possible to review all in a single article. In the current article we will turn our attention to those modalities that are either currently in use or in development for the medical imaging clinic. PMID- 14523979 TI - Real time non-invasive imaging of receptor-ligand interactions in vivo. AB - Non-invasive longitudinal detection and evaluation of gene expression in living animals can provide investigators with an understanding of the ontogeny of a gene's biological function(s). Currently, mouse model systems are used to optimize magnetic resonance imaging (MRI), positron emission tomography (PET), single-photon emission computed tomography (SPECT), and optical imaging modalities to detect gene expression and protein function. These molecular imaging strategies are being developed to assess tumor growth and the tumor microenvironment. In addition, pre-labeling of progenitor cells can provide invaluable information about the developmental lineage of stem cells both in organogenesis and tumorigenesis. The feasibility of this approach has been extensively tested by targeting of endogenous tumor cell receptors with labeled ligand (or ligand analog) reporters and targeting enzymes with labeled substrate (or substrate analog). We will primarily discuss MRI, PET, and SPECT imaging of cell surface receptors and the feasibility of non-invasive imaging of gene expression using the tumor microenvironment (e.g., hypoxia) as a conditional regulator of gene expression. PMID- 14523980 TI - Antisense imaging: and miles to go before we sleep? AB - Labeled oligonucleotide analogues for antisense imaging of messenger RNA (mRNA) have great potential for detection of endogenous gene expression in vivo. Successful antisense imaging may be useful for detecting cellular gene expression patterns and early molecular changes in disease. Conclusive demonstration of this technique has been hindered by formidable challenges in surmounting biological barriers and detecting low concentrations of target mRNA. Recent advances in the development of novel antisense molecules, high specific activity radiolabeling chemistry, sophisticated drug targeting technology, and complementary molecular imaging modalities make it quite possible that true antisense imaging will be realized in the near future. PMID- 14523981 TI - Molecular imaging in prostate cancer. AB - Prostate cancer (PCa) is the most common non-cutaneous malignancy in men. New ways to diagnose this cancer in its early stages are needed. Unique genetic and biochemical changes in the cell pave the way for tumors to grow and metastasize. Novel imaging approaches attempt to detect pathological processes in cancer cells at the molecular level. This has led to the establishment and development of the field of molecular imaging. Positron emission tomography (PET), magnetic resonance spectroscopic imaging (MRSI), magnetic resonance imaging (MRI), and radiolabeled antibodies are a few of the modalities that can detect abnormal tumor metabolic processes in the clinical setting. Other imaging techniques are still in their early phase of development but hold promise for the future, including bioluminescence imaging (BLI), measurement of tumor oxygenation, and measurement of uptake of iodine by tumors. These techniques are non-invasive and can spare the patient undue morbidity, while potentially providing early diagnosis, accurate follow-up and, finally, valuable prognostic information. PMID- 14523983 TI - Functional imaging of angiogenesis in an orthotopic model of pancreatic cancer. AB - Pancreatic cancer is a major unsolved health problem. The estimated overall 5 year survival rate of only 1-4% is due to aggressiveness of the disease and the lack of effective systemic therapies. Most pancreatic cancer-related deaths are due to the development of metastases, which represents the culmination of a complex interaction between the host organism and neoplastic cells within the primary tumor. Therefore, the study of tumor-host interaction in the context of the whole organism is necessary to evaluate the pathogenesis of tumor growth and metastasis so that effective therapies can be developed. Recent advances in functional imaging combined with animal models that faithfully recreate the biology of human tumors have elevated our ability to examine these complex interactions. In this review, we will use the example of orthotopic mouse models of pancreatic cancer as a tool to survey the challenges and possibilities of functional imaging of angiogenesis, a critical determinant of metastasis. PMID- 14523982 TI - Optical imaging and tumor angiogenesis. AB - Tumor angiogenesis is essential for tumor growth and progression. Therefore, targeting tumor blood vessels is a promising approach for cancer therapy. Angiogenesis, the formation of blood vessels, is a multistep process, and strongly influenced by the microenvironment. There are no in vitro assays that can resemble this dynamic process in vivo. For this reason, animal models and imaging technologies are critical for studying tumor angiogenesis, identifying therapeutic targets as well as validating the targets. Non-invasive molecular imaging in animal models presents an unprecedented opportunity and ability for us to perform repetitive observations and analysis of the biological processes underlying tumor angiogenesis and tumor progression in living animals in real time. As we gain a better understanding of the fundamental molecular nature of cancer, these techniques will be an important adjunct in translating the knowledge into clinical practice. This important information may elucidate how the tumor blood vessels behave and respond to certain treatments and therapies. PMID- 14523984 TI - Tunable, monochromatic X-rays: an enabling technology for molecular/cellular imaging and therapy. AB - Pulsed, tunable, monochromatic X-rays hold great potential as a cellular and molecular probe. These beams can be tuned to the binding energy of orbital electrons in atoms, making them extremely useful in diagnostic k-edge imaging and Auger cascade radiotherapy. Their wide tunability makes them ideal for the performance of various techniques as disparate as protein crystallography and three-dimensional, compressionless, monochromatic mammography. Since only the frequency best suited to the task at hand is used, radiation exposure to patients or animals is exceedingly low when compared to standard X-ray techniques. PMID- 14523985 TI - Combinatorial discovery of tumor targeting peptides using phage display. AB - Peptides possess appropriate pharmacokinetic properties to serve as cancer imaging or therapeutic targeting agents. Currently, only a small number of rationally-derived, labeled peptide analogues that target only a limited subset of antigens are available. Thus, finding new cancer targeting peptides is a central goal in the field of molecular targeting. Novel tumor-avid peptides can be efficiently identified via affinity selections using complex random peptide libraries containing millions of peptides that are displayed on bacteriophage. In vitro and in situ affinity selections may be used to identify peptides with high affinity for the target antigen in vitro. Unfortunately, it has been found that peptides selected in vitro or in situ may not effectively target tumors in vivo due to poor peptide stability and other problems. To improve in vivo targeting, methodological combinatorial chemistry innovations allow selections to be conducted in the environment of the whole animal. Thus, new targeting peptides with optimal in vivo properties can be selected in vivo in tumor-bearing animals. In vivo selections have been proven successful in identifying peptides that target the vasculature of specific organs. In addition, in vivo selections have identified peptides that bind specifically to the surface of or are internalized into tumor cells. In the future, direct selection of peptides for cancer imaging may be expedited using genetically engineered bacteriophage libraries that encode peptides with intrinsic radiometal-chelation or fluorescent sequences. PMID- 14523986 TI - Molecular magnetic resonance imaging with targeted contrast agents. AB - Magnetic resonance imaging (MRI) produces high-resolution three-dimensional maps delineating morphological features of the specimen. Differential contrast in soft tissues depends on endogenous differences in water content, relaxation times, and/or diffusion characteristics of the tissue of interest. The specificity of MRI can be further increased by exogenous contrast agents (CA) such as gadolinium chelates, which have been successfully used for imaging of hemodynamic parameters including blood perfusion and vascular permeability. Development of targeted MR CA directed to specific molecular entities could dramatically expand the range of MR applications by combining the noninvasiveness and high spatial resolution of MRI with specific localization of molecular targets. However, due to the intrinsically low sensitivity of MRI (in comparison with nuclear imaging), high local concentrations of the CA at the target site are required to generate detectable MR contrast. To meet these requirements, the MR targeted CA should recognize targeted cells with high affinity and specificity. They should also be characterized by high relaxivity, which for a wide variety of CA depends on the number of contrast-generating groups per single molecule of the agent. We will review different designs and applications of targeted MR CA and will discuss feasibility of these approaches for in vivo MRI. PMID- 14523987 TI - Choline phospholipid metabolism: a target in cancer cells? AB - The experience of treating cancer over the past several decades overwhelmingly demonstrates that the disease continues to evade the vast array of drugs and treatment modalities available in the twenty-first century. This is not surprising in view of the complexity of this disease, and the multiplicities of pathways available to the cancer cell to enable its survival. Although the progression of cancer arrives at a common end point of cachexia, organ failure, and death, common pathways are rare in cancer. Identifying and targeting common pathways that would act across these levels of multiplicity is essential for the successful treatment of this disease. Over the past decade, one common characteristic consistently revealed by magnetic resonance spectroscopic studies is the elevation of phosphocholine and total choline-containing compounds in cancer cells and solid tumors. This elevation has been observed in almost every single cancer type studied with NMR spectroscopy and can be used as an endogenous biomarker of cancer. In this article, we have summarized some of the observations on the choline phospholipid metabolism of cancer cells and tumors, and make a case for targeting the aberrant choline phospholipid metabolism of cancer cells. PMID- 14523988 TI - Hpr6.6 protein mediates cell death from oxidative damage in MCF-7 human breast cancer cells. AB - Reactive oxygen species (ROS) cause cell death and are associated with a variety of maladies, from trauma and infection to organ degeneration and cancer. Cells mount a complex response to oxidative damage that includes signaling from transmembrane receptors and intracellular kinases. We have analyzed the response to oxidative damage in human breast cancer cells expressing the Hpr6.6 (human membrane progesterone receptor) protein. Although Hpr6.6 is related to a putative progesterone-binding protein, Hpr6.6 is widely expressed in epithelial tissues and shares close homology with a budding yeast damage response protein called Dap1p (damage response protein related to membrane progesterone receptor). We report here that the Hpr6.6 protein regulates the response to oxidative damage in breast cancer cells. Expression of Hpr6.6 in MCF-7 cells sensitized the cells to death following long-term/low dose or short-term/high dose treatment with hydrogen peroxide. Cell death did not occur through a typical apoptotic mechanism and corresponded with hyperphosphorylation of the Akt and IkappaB proteins. However, inhibition of Akt activation and IkappaB degradation had no effect on Hpr6.6-mediated cell death, suggesting that Hpr6.6 regulates cell death through a novel oxidative damage response pathway. Our work indicates a key regulatory function for Hpr6.6 in epithelial tissues exposed to oxidative damage. PMID- 14523989 TI - Cyclin dependent kinase inhibitor p27(Kip1) is upregulated by hypoxia via an ARNT dependent pathway. AB - Expression of cyclin dependent kinase (Cdk) inhibitor p27(Kip1), which blocks cell cycle progression from G(1) to S phase, can be regulated via multiple mechanisms including transcription, protein degradation, and translation. Recently, it was shown that p27(Kip1) plays an important role in the cellular response to hypoxia. However, the mechanisms involved in the hypoxia-induced regulation of p27(Kip1) expression are still not clear. In this study, we compare the expression of p27(Kip1) in two related murine hepatoma cell lines, Hepa-1 and c4. Hepa-1 produces functional aryl hydrocarbon receptor nuclear translocator (ARNT). c4 cells are derived from Hepa-1, but are ARNT deficient. Interestingly, we observed cell line-dependent effects of hypoxia on the expression of p27(Kip1). The level of p27(Kip1) protein in Hepa-1 cells is enhanced by hypoxia, but is reduced by hypoxia in c4 cells. Further investigation demonstrated that hypoxia-induced, ARNT-mediated, transactivation of the p27(Kip1) gene in Hepa-1 cells is responsible for the increase in p27(Kip1) protein. Once c4 cells were stably transfected with the wild type ARNT gene, a hypoxia-induced increase in p27(Kip1) mRNA was observed and reduction of p27(Kip1) protein caused by hypoxia was blocked. Hence, our data indicate that ARNT is involved in transcriptional upregulation of the p27(Kip1) gene under hypoxic conditions. PMID- 14523990 TI - ErbB2 and EGFR are downmodulated during the differentiation of 3T3-L1 preadipocytes. AB - The expression of receptors belonging to the epidermal growth factor receptor subfamily has been largely studied these last years in epithelial cells mainly as involved in cell proliferation and malignant progression. Although much work has focused on the role of these growth factor receptors in the differentiation of a variety of tissues, there is little information in regards to normal stromal cells. We investigated erbB2 expression in the murine fibroblast cell line Swiss 3T3L1, which naturally or hormonally induced undergoes adipocyte differentiation. We found that the Swiss 3T3-L1 fibroblasts express erbB2, in addition to EGFR, and in a quantity comparable to or even greater than the breast cancer cell line T47D. Proliferating cells increased erbB2 and EGFR levels when reaching confluence up to 4- and 10-fold, respectively. This expression showed a significant decrease when growth-arrested cells were stimulated to differentiate with dexamethasone and isobutyl-methylxanthine. Differentiated cells presented a decreased expression of both erbB2 and EGFR regardless of whether the cells were hormonally or spontaneously differentiated. EGF stimulation of serum-starved cells increased erbB2 tyrosine phosphorylation and retarded erbB2 migration in SDS-PAGE, suggesting receptor association and activation. Heregulin-alpha1 and beta1, two EGF related factors, had no effect on erbB2 or EGFR phosphorylation. Although 3T3-L1 cells expressed heregulin, its specific receptors, erbB3 and erbB4, were not found. This is the first time in which erbB2 is reported to be expressed in an adipocytic cell line which does not depend on non EGF family growth factors (thyroid hormone, growth hormone, etc.) to accomplish adipose differentiation. Since erbB2 and EGFR expression were downmodulated as differentiation progressed it is conceivable that a mechanism of switching from a mitogenic to a differentiating signaling pathway may be involved, through regulation of the expression of these growth factor receptors. PMID- 14523991 TI - Analysis of SOX10 mutations identified in Waardenburg-Hirschsprung patients: Differential effects on target gene regulation. AB - SOX10 is a member of the SOX gene family related by homology to the high-mobility group (HMG) box region of the testis-determining gene SRY. Mutations of the transcription factor gene SOX10 lead to Waardenburg-Hirschsprung syndrome (Waardenburg-Shah syndrome, WS4) in humans. A number of SOX10 mutations have been identified in WS4 patients who suffer from different extents of intestinal aganglionosis, pigmentation, and hearing abnormalities. Some patients also exhibit signs of myelination deficiency in the central and peripheral nervous systems. Although the molecular bases for the wide range of symptoms displayed by the patients are still not clearly understood, a few target genes for SOX10 have been identified. We have analyzed the impact of six different SOX10 mutations on the activation of SOX10 target genes by yeast one-hybrid and mammalian cell transfection assays. To investigate the transactivation activities of the mutant proteins, three different SOX target binding sites were introduced into luciferase reporter gene constructs and examined in our series of transfection assays: consensus HMG domain protein binding sites; SOX10 binding sites identified in the RET promoter; and Sox10 binding sites identified in the P0 promoter. We found that the same mutation could have different transactivation activities when tested with different target binding sites and in different cell lines. The differential transactivation activities of the SOX10 mutants appeared to correlate with the intestinal and/or neurological symptoms presented in the patients. Among the six mutant SOX10 proteins tested, much reduced transactivation activities were observed when tested on the SOX10 binding sites from the RET promoter. Of the two similar mutations X467K and 1400del12, only the 1400del12 mutant protein exhibited an increase of transactivation through the P0 promoter. While the lack of normal SOX10 mediated activation of RET transcription may lead to intestinal aganglionosis, overexpression of genes coding for structural myelin proteins such as P0 due to mutant SOX10 may explain the dysmyelination phenotype observed in the patients with an additional neurological disorder. PMID- 14523992 TI - Modification of phospholipids fatty acid composition in reuber H35 hepatoma cells: effect on HMG-CoA reductase activity. AB - There is controversy about the effect of saturated and polyunsaturated fats on 3 hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase, the main regulatory enzyme of cholesterogenic pathway. Results from dietary studies are difficult to interpret because diets normally contain a mixture of fatty acids. Therefore, we have used Reuber H35 hepatoma cells whose phospholipids were enriched in different individual fatty acids and have studied their effects on the cellular reductase activity. Lauric, myristic, eicosapentaenoic (EPA), and docosahexaenoic (DHA) acids were supplemented to the culture medium coupled to bovine serum albumin. The four fatty acids were incorporated into phospholipids from cells grown in media containing whole serum or lipoprotein-poor serum (LPPS). Reductase activity of cells cultivated in a medium with LPPS was three to four times higher than those cultivated in medium with whole serum. Saturated fatty acids increased reductase activity of cells grown in medium with whole serum, whereas n-3 polyunsaturated fatty acids (PUFA) decreased it. However, both saturated and polyunsaturated fatty acids increased reductase activity when serum lipoproteins were removed. In conclusion, this is one of the first reports demonstrating that saturated and n-3 PUFA only show differential effects on HMG-CoA reductase activity in the presence of lipoproteins. PMID- 14523993 TI - Two alanines juxtaposed to aggrecan's G1 domain alter its intracellular localization. AB - Nascent proteins translated and processed in the endoplasmic reticulum (ER) sometimes contain intrinsic signals for ER retention or ER retrieval. These signals are usually a few amino acids in length, and if alanine modifications are made within these sequences, normal transit patterns of the nascent protein frequently change. The purpose of this study was to determine whether two alanines juxtaposed to the first globular domain of aggrecan's core protein affect its transit in Chinese hamster ovary (CHO) cells. Results show that two alanines juxtaposed to the first globular domain (G1AA) minimized secretion of the protein. However, transgenic proteins with juxtaposed glutamate-phenylalanine (G1EF) or no additional amino acids (G1) were still secreted. GFP-tagged G1AA localized in the lumen of the ER but not in the Golgi. In contrast, a portion of GFP-tagged G1EF and G1 did appear in the Golgi compartment. More importantly, unique and striking accumulations of G1EF and G1 transgenic proteins were seen in large dilated regions of the ER cisternae, reminiscent of accumulations seen in alpha1-antitrypsin deficiency disease. G1AA transgenic proteins did not form these vesicles but were diffusely distributed throughout the ER lumen. These results indicate that just two juxtaposed alanines can profoundly affect a large globular protein's intracellular localization. PMID- 14523994 TI - Identification of gammaA-like protocadherin expressed during chick development. AB - The protocadherins are calcium-dependent cell adhesion molecules of the cadherin superfamily that have been described in numerous species. Although less well characterized than classical cadherins, the protocadherins are also thought to facilitate critical cell-cell interactions during embryonic development. We have cloned a novel protocadherin from the embryonic chick utilizing a monoclonal antibody produced against a peanut agglutinin-binding fraction of cultured chick limb tissue to screen a lambdaZAP cDNA expression library from the stage 25 limb. A 2.8 kb cDNA clone was obtained that encoded multiple cadherin-like ectodomains. Northern blotting revealed a single 4.6 kb RNA transcript that was highly enriched in the stage 43 chick brain. Utilization of 3' Rapid amplification of cDNA ends (RACE) identified the entire 2.4 kb reading frame. The chick protocadherin contained five cadherin-like extracellular repeats and a highly conserved cytoplasmic domain. Amino acid alignment of the extracellular domains revealed marked identity to the human gammaA protocadherin subfamily. In situ hybridization showed low levels of mRNA localization in several developing chick tissues, but stronger expression in the neural tube and dorsal root ganglia at stage 27. In the stage 43 chick brain, protocadherin mRNA was noted in discrete regions, particularly within the developing optic lobe. As for protocadherins described in other species, these results suggest that this novel gammaA-like protocadherin may also play a role in chick neural development. PMID- 14523995 TI - Overexpression of regucalcin modulates tumor-related gene expression in cloned rat hepatoma H4-II-E cells. AB - Regucalcin is a regulatory protein in intracellular signaling pathway which is related to various protein kinases and protein phosphatases in many cells. The effect of regucalcin on the expression of tumor-related genes was investigated in the cloned rat hepatoma H4-II-E cells and the hepatoma cells (transfectants) overexpressing regucalcin. Hepatoma cells were cultured for 24-72 h in the presence of fetal bovine serum (FBS; 10%). The proliferation of hepatoma cells was significantly suppressed at 24-72 h of culture in regucalcin transfectants as compared with that of wild-type or mock-type cells. Western blot analysis showed that regucalcin was markedly expressed in transfectants. The expression of c-myc, c-fos, c-jun, Ha-ras, and p53 mRNAs was determined using reverse transcription polymerase chain reaction (RT-PCR). Of these genes, the expression of c-myc or Ha ras mRNAs was significantly suppressed in regucalcin transfectants. The suppression of c-myc mRNA expression in transfectants was confirmed by using Northern blot analysis; significant suppression was seen at 24, 48, or 72 h of culture in the presence of 10% FBS. Culture with 10% FBS significantly enhanced c myc mRNA expression in the hepatoma cells (wild-type) as compared with that of 1% FBS. The enhancement was significantly abolished in the transfectants. Meanwhile, the expression of p53 mRNA in the hepatoma cells was significantly enhanced in regucalcin-overexpressing hepatoma cells. This study demonstrates that the expression of oncogene c-myc and Ha-ras mRNA in hepatoma cells overexpressing regucalcin is suppressed, and that the tumor suppression gene p53 is enhanced in the transfectants. PMID- 14523996 TI - Sodium arsenite downregulates transcriptional activity of AP-1 and CRE binding proteins in IL-1beta-treated Caco-2 cells by increasing the expression of the transcriptional repressor CREMalpha. AB - In recent studies, sodium arsenite (SA) inhibited IL-6 production in cultured intestinal epithelial cells, at least in part by downregulating the activity of nuclear factor-kappaB (NF-kappaB). The influence of SA on the activity of other transcription factors regulating the interleukin-6 (IL-6) gene in enterocytes is not known. We tested the effect of SA on the activity of CCAAT/enhancer binding protein (C/EBP), activating protein-1 (AP-1), and CRE binding proteins in IL 1beta-treated Caco-2 cells. DNA binding activity was determined by electrophoretic mobility shift assay (EMSA) and transcriptional activity by transfecting cells with luciferase reporter plasmids containing promoter constructs with binding sites for the individual transcription factors. DNA binding activity for all three transcription factors was increased after treatment with SA or IL-1beta. In contrast, SA inhibited transcriptional activity of AP-1 and CRE binding proteins but not C/EBP. Additional experiments provided evidence that the inhibition of AP-1 and CRE mediated transcriptional activity was associated with, and probably caused by, increased expression of the transcriptional repressor cyclic AMP response element modulator (CREM)alpha. The present results are consistent with the concept that SA inhibits IL-6 production in stimulated enterocytes by downregulating the transcriptional activity of several, but not all, IL-6-related transcription factors. Because of the multiple important biological functions of IL-6 in the enterocyte and gut mucosa, methods to regulate enterocyte IL-6 production have significant clinical implications. PMID- 14523998 TI - AB0 blood subgroup allele frequencies in the Turkish population. AB - We determined the AB0 blood group system with a PCR based technique termed APLP (Amplified Product Length Polymorphism) in the Turkish population. The method includes ten different allele specific primers and permits identification of the major AB0 genotypes and its suballeles (A1-A2-B-0A-02-0G-AG). The suballeles were amplified in a single tube reaction. We have determined AB0 phenotypes in 129 Turkish individuals. No significant deviation from the Hardy-Weinberg equilibrium was observed. PMID- 14523997 TI - Profibrillin-1 maturation by human dermal fibroblasts: proteolytic processing and molecular chaperones. AB - Fibrillin-1 is synthesized as a proprotein that undergoes proteolytic processing in the unique C-terminal domain by a member of the PACE/furin family of endoproteases. This family of endoproteases is active in the trans-Golgi network (TGN), but metabolic labeling studies have been controversial as to whether profibrillin-1 is processed intracellularly or after secretion. This report provides evidence that profibrillin-1 processing is not an intracellular event. Bafilomycin A(1) and incubation of dermal fibroblasts at 22 degrees C were used to block secretion in the TGN to confirm that profibrillin-1 processing did not occur in this compartment. Profibrillin-1 immunoprecipitation studies revealed that two endoplasmic reticulum-resident molecular chaperones, BiP and GRP94, interacted with profibrillin-1. To determine the proprotein convertase responsible for processing profibrillin-1, a specific inhibitor of furin, alpha-1 antitrypsin, Portland variant, was both expressed in the cells and added to cells exogenously. In both cases, the inhibitor blocked the processing of profibrillin 1, providing evidence that furin is the enzyme responsible for profibrillin-1 processing. These studies delineate the secretion and proteolytic processing of profibrillin-1, and identify the proteins that interact with profibrillin-1 in the secretory pathway. PMID- 14523999 TI - Genetic diversity of serum proteins in three subpopulations of the Maria Gond tribe of Madhya Pradesh, India. AB - The phenotype and allele frequency distribution of group specific component (GC), transferrin (TF), alpha-1-antitrypsin (PI) and apolipoprotein E (APOE) was determined by isoelectric focusing of plasma samples from three subpopulations (Bison Horn Maria of the Kuakonda and Tokapal Block, and Abuj Maria of the Abujmar Hills of the Orchha block) of the Maria Gond tribe of Madhya Pradesh, India. A considerable level of allele frequency variation was observed in these subpopulations, which highlighted social and geographical isolation among them. The average heterozygosity for these IEF subtype systems was high (29-39%) and the gene diversity among these subpopulation groups was of low to moderate range (1.4%). The overall analysis showed that these polymorphisms are useful anthropological markers for micro-evolutionary and genetic structure studies. PMID- 14524000 TI - Genetic studies among seven endogamous populations of the Koshi Zone, Bihar (India). AB - The distribution of AB0 and Rhesus blood groups, PTC taste sensitivity and colour blindness was studied among seven endogamous populations (Tharu, Mushar, Santal, Dhobi, Julaha, Kulhaiya and Karan Kayastha) in the Koshi Zone of Bihar (India). The phenotype and allele frequencies of the four gene loci (AB0, RH, PTC and colour blindness) show considerable differences between these populations. The measurement of genetic distances revealed, that the lowest genetic distance is seen between Dhobi and Julaha, the highest between Mushar and Tharu. From the genetic distance analysis there is some evidence for a close genetic relationship among the population groups belonging to the same region, irrespective of their caste, religion, linguistic or any other affinities. It may be concluded that all these populations have arisen through a common ancestor and changed gene frequencies among them is due to evolutionary forces like mutation, selection, migration, temporal variation and genetic drift. However, these populations retain their separate entities by practising endogamy. Gene diversity analysis reveals that these populations are at an early stage of genetic differentiation. PMID- 14524001 TI - Morphological and regional variations in body dimensions of the Gujjars of different localities in north-western India. AB - Physical features and somatometric characters of the wide spread Gujjar population in North-Western India are fascinating. It is all the more interesting to study their morphological and regional variations in the body dimensions. In this paper, anthropometric characters of 200 adult subjects each belonging to Hindu Gujjars in the Ropar District of Punjab and Muslim Gujjars in the Chamba District of Himachal Pradesh in the North-Western part of India were compared. They differ significantly from each other for many physiognomic measurements, suggesting the biological diversity between the two population groups. Further, the comparison of anthropometric measurements between the different localities of the Gujjars showed morphological variations and regional diversity of the isolates in North-Western India. These findings may be attributed to the founder effect, genetic drift, and breeding and geographical isolation of the populations under study and not to secular trends. The findings have also been compared and discussed with the available results of other local populations in North-Western India. PMID- 14524002 TI - The question of differences in body dimensions between children from endogamic and exogamic matings: a study in Sardinia. AB - The aim of this study is to investigate whether there are differences in body dimensions between children of the same sex from endogamic and exogamic matings. The cross-sectional sample consisted of 285 children, 136 males and 149 females, 6 to 10 years old, attending elementary schools in Tortoli, a town in east central Sardinia. The children of each sex were divided into two groups: endogamic and exogamic, the first including children from parents born in the same Sardinian village, the latter children from parents born in two different places. For each sex, ANCOVA with age as covariate revealed no significant differences between the two groups of children for the 35 anthropometric variables considered. Moreover, for each sex, there were no significant differences between the two groups of children for some anthropometric variables considered to be indicators of nutritional status: sums of skinfold thicknesses, waist/hip ratio, body mass index, total upper arm area, upper muscle arm area, upper arm fat area. We conclude that when Sardinian children of endogamous and exogamous unions are in similar nutritional conditions, they do not differ in body dimensions. PMID- 14524003 TI - The impact of the weight status on subjective symptomatology of the Polycystic Ovary Syndrome: a cross-cultural comparison between Brazilian and Austrian women. AB - Group differences in symptomatology and symptom perception of young women ageing between 18 and 32 years suffering from polycystic ovarian syndrome (PCOS) were tested according to cultural background and weight status. In detail 31 Austrian women, living in Carinthia, Austria, and 102 Brazilian women, living in Sao Paulo, Brazil, were enrolled in the present study. All participants suffered from diagnosed PCOS. The prevalence of hirsutism, infertility, menstrual disturbances and overweight/obesity and their individual impact on health related quality of life were analysed. Furthermore the impact of weight status (BMI) on symptom perception was tested. It turned out, that the Brazilian sample exhibited higher prevalence of PCOS symptoms and these symptoms had a more negative impact on quality of life. The only exception was body weight. Although significantly leaner, the Austrian women showed a higher Cronin score of body weight than their Brazilian counterparts. The results of the present study may indicate that in western industrialized societies the fear of overweight is much more prevalent than in a developing country such as Brazil and thus it has more influence on the quality of life than all the other symptoms. PMID- 14524004 TI - Age variations in adiposity and body fat composition among older Bengalee Hindu women of Calcutta, India. AB - A cross-sectional study of 279 older (50+ years) urban Bengalee Hindu women was undertaken to study age variations in adiposity, body composition, obesity and central fat distribution. The women were divided into three groups: Group I (G I, 50-59 years), Group II (G II, 60-69 years) and Group III (G III, 70+ years). A significant decreasing age trend was observed in adiposity and body fat composition measures. Women in G I had significantly higher means compared with those in G III. Individuals in G II had intermediate values. However, there was no significant age trend in muscle measures and indices of central body fat distribution. The results revealed that significantly more women in G III (45.8%) were malnourished (BMI < 18.5), while significantly more women in G I (28.7%) were obese (BMI > or = 25). The levels of malnourishment (21.6%) and obesity (24.5%) in G II were intermediate between G I and G III. Age had significant negative correlations with measures of adiposity and body fat composition. Regression analysis revealed that age had significant negative effect on these anthropometric measures. This significant negative impact of age remained even after controlling for the effect of BMI. In conclusion, the present investigation revealed that among older Bengalee Hindu women, there is a significant inverse age trend in adiposity and body fat composition, which is independent of overall adiposity (BMI). However, with ageing, muscle and central body fat distribution remain the same. Furthermore, with increasing age, there is a trend of increasing levels of malnourishment and decreasing levels of obesity. PMID- 14524005 TI - Association between immunoglobulin allotypes and pulmonary tuberculosis. AB - In a sample of n = 133 non-related patients suffering from pulmonary tuberculosis, Gm and Km typings have been carried out and compared with healthy controls from the same geographical area. All the Gm allotypes tested were found to be more preponderant in the patients than in the healthy controls and these differences were found to be statistically significant for Gm (1) and Gm (5) only and not for the other immunoglobulin allotypes e.g. Gm (2). The frequency of Km (1) was lower and that of Km (3) was higher in the patients than in the controls. These differences were, however, statistically not significant. PMID- 14524006 TI - Results of an explorative empirical study on human mating in Germany: handsome men, not high-status men, succeed in courtship. AB - Recent research on human mating depicts men as searching for physical attractiveness (PA) and women as searching for status. To identify the mechanisms which lead to universal, biologically interpretable structures in social processes, we focused on the proximate causes for inter- and intrasexual differences in human mating preferences, attraction, and tactics. We collected data on 180 young singles (mean age 26.9 years) without a steady relationship. A questionnaire and a video sequence (20-30 seconds) of each subject was taken. Next, each video sequence was rated by approximately 20 individuals of the opposite sex, who also participated in this study. Surprisingly, the answers given by male and female subjects regarding sociosexual behaviour and mating preferences are predominantly congruent. Sex differences among preferences for good looking and high-status partners were small or even insignificant. Lower educated subjects had considerably higher status preferences than higher educated individuals. In both sexes, PA was much more preferred in a potential partner than status. For both sexes, physical appearance was decisive for the subject's dating attractiveness. Male, but not female dating attractiveness also correlates with a kind and charismatic appearance. Furthermore, there was a positive linear relationship between men's PA and their number of sexual partners within the last year. Men with more than four sexual partners were all above-average in PA, while the most attractive women had a medium number of sexual partners. However, in this respect, status had no influence. The results show that sex differences in mating are more complex than hitherto assumed. PMID- 14524007 TI - [Effect of premedication on the formation of postoperative pain syndrome]. AB - The pronouncement degree and dynamics of pain syndrome were assessed after surgeries in the abdominal cavity organs with regard for a nature of the analgetic component of premedication and of the postoperative analgesic therapy (n = 155). The administration of analgesics before extra-traumatic surgeries was found to be compulsory in order to prevent the onset of a persistent postoperative pain syndrome. A combination of non-steroid anti-inflammatory drugs, i.e. ketonal, and of a high-efficiency agonist of opiate mu-receptors, i.e. bupranal, was acknowledged as most effective in coping with this task. It was demonstrated that the application of "the forestalling analgesia" by ketonal (100 mg in 30 min before the beginning of moderate- and mild-trauma surgical interventions and immediately after the finalization of such surgeries) contributed to an essentially lower pronouncement degree of the postoperative pain syndrome and made the administration of narcotic analgesics unnecessary. Finally, it is pointed out that promedol could not be regarded as a drug ensuring a safe prophylaxis of the central sensitization and hyperalgesia. PMID- 14524008 TI - [Assessment of anesthesia adequacy with spectral analysis of the heart sinus rhythm]. AB - The information density of spectral analysis of the variability of sinus rhythm to be used in diagnosing the neurovegetative activity in operation trauma was studied in 32 patients in the course of anesthesia and operation. It is suggested on the basis of the found the method can in monitoring the anesthesia adequacy. PMID- 14524009 TI - [Prevention of altering effects of surgical stress in children with abdominal pathology]. AB - The controlled activation of endogenous stress-limiting systems by using metabolites, their synthetic analogues and natural antioxidants was shown, within the case study, to prevent a excessive tension of the sympathetic-and-adrenalin systems and to ensure an adequate anesthetic protection against the surgical stress. PMID- 14524010 TI - [Characteristics of hemodynamic reactions during various endoscopic surgeries for cholelithiasis in patients with concurrent cardiovascular pathologies]. AB - A prospective study of 170 patients with concurrent cardiac pathology, who had underwent the laparoscopic cholecystectomy under the conditions of a tense carboxyperitomium, led to isolating two groups of patients with high-risk of intra- and postoperative unfavorable cardiac-vascular responses: an original high risk degree (III-IV classes according to ASA), a high value of cardiac risk indices according to Goldman (III-IV classes) and according to Detsky (II-III classes), a hypodynamic blood circulation regimen under rest conditions, and ejection fraction (according to echocardiography) amounting below 50%. Such patients are recommended to be operated on by laparolifting or by low-gas laparoscopy with a local lifting, i.e. by using the surgical techniques free of any negative effects produced on the carboxyperitomium hemodynamics, while all positive features of small-invasion surgery remain intact. PMID- 14524011 TI - [Effect of epidural morphine analgesia on the NO-forming activity of nociceptive neurons of the spinal ganglia and bone marrow]. AB - The effect of epidural morphine analgesia on NO mechanisms was studied during the primary and secondary sensitizations. The nitro-oxide synthase (NOS) activity was evaluated by the method of Home and Vincent, 1989, (identification of NADPH diaphorase). NOS and NADPH diaphorases are two double enzymes in cells with a synchronously changing activity. The NOS activity is in line with the level of the NO secretion. The enzyme was measured on a microdensitometer. The findings evidence an active involvement of NO mechanisms in the formation of primary and secondary hyperalgesia, and the injection of morphine caused a significant decrease in the count of NO-expressing neurons in the total number of cells with high and average activities of the enzyme. PMID- 14524012 TI - [Evaluation of the local neurotoxicity of lidocaine following epidural administration]. AB - The morphological and histoenzymological changes in spinal neuron and in the spinal ganglia of outbred dogs were analyzed in the epidural administration of 2% solution of lydocaine. The results showed a lack of pathological or structural and-metabolic changes in the nervous tissue. The morpho-functional reshaping occurring in the spinal cord cells and in the spinal ganglia was in line with that when the 0.9% solution of sodium chloride was in use. The registered changes ranging within the physiological deviations' limits witnessed to a functional reaction of the studied nervous-tissue structures to epidural drugs' injections. PMID- 14524013 TI - [Use of epidural analgesia for the arresting of pronounced lumbar pains]. AB - A combination of local anesthetics (2% lydokaine solution, 2 mg/kg; 0.5% bupivakaine solution, 0.4 mg/kg or 0.75% ropivakaine solution, 0.55 mg/kg), promedol (20 mg) and corticosteroids (125 mg hydrocortisone or 80 mg kenalog) was epidurally administered in 78 patients to arrest the pronounced root pain syndrome of the lumbar-sacral localization provoked by intervertebral disk hernias. The most prolonged analgesic effect was registered in groups, whose patients received bupivakaine or ropivakaine, which prevented the onset of pain before the antiinflammatory effect of corticosteroids started. The patient must be informed well in advance about the possibility of a momentary relapse of pains and about the intensification (at the same time moment) of pharmacotherapy prescribed by neuropathologist. PMID- 14524014 TI - [Development of the Russian standards (protocols) for basic and advanced resuscitation complexes]. AB - Serious organizational gaps related with the practice of cardiac-pulmonary reanimation still persist in Russia until now. Therefore, it is suggested to work out unified state standards of primary and advanced reanimation complexes and training schemes in cardiac-pulmonary reanimation for medical and other population categories as well as the stage-by-stage dissemination of such standards throughout the country's territory. PMID- 14524015 TI - [Interhospital transportation of children from paediatric intensive care units within a major region]. AB - The activity of a regional reanimation-and-consultation center (RCC) related with transportation of 3219 children (79.5% of them were newborns) is summarized in the paper. The share of children assigned for artificial lung ventilation (ALV) increased from 50.8% to 82.6%, while the lethality rate dropped from 12.8% to 6.9%. The above results were enabled by a maximally early re-hospitalization of patients from therapeutic-and-prevention facilities (TPF) with limited medical care abilities and by early preparations (for transportation) by using the DINAR 2 expert system. The children were transported only after the main vitally important systems had been compensated for and with support of an adequate intensive care therapy. Should the above principles be met, the specialized therapy could be started at he earliest possible time; it promoted the therapy quality and cut the patients' stay in intensive care units. PMID- 14524016 TI - [Effectiveness of oxygen therapy for pulmonary edema in patients with hypertensive crisis before hospitalization]. AB - The oxygen therapy, involving the KI-3M respiratory apparatus administered in the positive pressure regimen on expiration (8-10 cm aq. c.), used, before hospitalization, within the complex therapy of patients with pulmonary edema triggered by hypertensive crisis arrests fast enough the pulmonary edema, cuts the working time of an ER crew and promotes the ER efficiency. The key clinical manifestations of pulmonary edema were found to begin to go down on minutes 5 to 10 after the administration of artificial lungs ventilation (ALV) by oxygen with the positive pressure being on expiration end; such signs were totally stopped in 20-25 minutes. PMID- 14524018 TI - [The modern problems of intensive care in patients with wounds and traumas]. AB - The results of rendering the intensive care to 2244 patients with severe wounds and traumas were analyzed, which made it possible to define the most topical problems needed to be resolved, i.e. treatment schemes for patients with vegetative state (similar to coma) and with asthenic syndrome, perfection of intensive care for the elderly and absolutization of some intensive care techniques. Perspective solutions are suggested to cope with the above issues, which comprise a bigger quantity of rehabilitation centers, an elaboration of pathogenetic tactics of intensive care for the elderly patients and revising the feasibility of a long-term use of the intensive care methods with respect to the regulation and integration suppressed by such methods. PMID- 14524017 TI - [New methods of prevention and treatment of the stress-related disorders in children with burn disease]. AB - An assessment of the key pathogenesis chains involved in triggering the vital complications was based on a prospective randomized controllable retrospective study of a burn-disease clinical course in 473 children. As a result of it, a standardized system was worked out, within a large industrial region, designed to render the medical care to children with severe thermal lesion; such system cut the therapy time by 12 days in the intensive care unit and by 2.6 time--in hospital; it also contributed to reducing the purulent-and-septic complications from 73.9% to 21% and the lethality rate--from 3% to 0.8%. PMID- 14524019 TI - [Monitoring of extravascular lung water in patients with severe sepsis]. AB - The key objective of the case study was the possibility to monitor the extravascular lung water (EVLW) in severe cases. Twelve mechanically ventilated patients with severe sepsis complicated by septic shock and by an acute lung injury (ALI) were involved in the prospective study. The measurements, performed on days 1 and 3 after the onset of sepsis, comprised hemodynamics, EVLW as assessed by Pulsion PiCCO method, blood gases and severity scores. The EVLW correlated significantly with lung injury score (r = 0.46), oxygenation (r = 0.46) and with pulmonary compliance (r = -0.58) versus the central venous pressure. The EVLW and lung injury scores were found to be essentially higher in non-survivors on day 3. The clinical situations, described in the present article, are indicative of a potential EVLW value applicable to sepsis treatment. Finally, the monitoring of EVLW is a useful tool in the purpose-oriented therapy of sepsis-induced ALI; moreover, the method has an important prognostic value. PMID- 14524020 TI - [Psychoemotional status of patients with endogenous intoxication in peritonitis]. AB - While holding a complex neuropsychological testing of 60 patients with disseminated peritonitis, we encountered a problem of a secondary reactive somatopsychosis due to metabolic encephalopathy. It is noteworthy, that the severity of psychoemotional disorders expectedly progressed depending on a degree of endogenous intoxication. Such circumstance must be duly considered in composing and implementing an intensive care scheme for the discussed patients' category. PMID- 14524021 TI - [Clinical and electrophysiological approaches towards diagnostics and therapy of patients in vegetative state]. PMID- 14524022 TI - [Optimization of intensive care of acute disseminated intravascular syndrome]. AB - The discrete plasmapheresis and Fraxiparin were used to optimize the intensive care scheme in acute disseminated intravascular syndrome (DIS). A total of 394 patients underwent the clinical and laboratory examinations to assess the method's efficiency. A lower degree thrombinemia was registered after the therapy versus the original values. PMID- 14524023 TI - [Natural surfactant preparation in complex therapy of acute respiratory distress syndrome in adults following open-heart surgeries]. AB - The clinical course and outcomes of cases of acute respiratory distress syndrome (ARDS) as evolved after open-heart surgeries under the conditions of traditional treatment and under the conditions of using the exogenous surfactant drug were comparatively analyzed. A reliably improved oxygenation was registered in the 1st group patients (18 subjects), who received surfactant, yet 24 hours later; while the identical improvement was noted in the 2nd group patients (19 subjects) only in 98 hours. The possibility of respiratory support with FiO2 below 45% was ensured in 15 (83.3%) patients of group 1 and in 10 (52.6%) patients of group 2. Thirteen (72.2%) patients in the 1st group and 9 (47.4%) patients in the 2nd group were transferred to independent breathing. The lethality rate in ARDS with the surfactant therapy made up 33.3%, it was 52.6% in traditional treatment. PMID- 14524024 TI - [Variants of the course of early noncomplicated postoperative period following excision of hypothalamo-hypophysial region tumors in children]. AB - A study of non-specific and specific reactions in neurosurgical patients as conducted in the early postoperative period revealed three main variations of an early postoperative clinical course, i.e. with a normal stress-reaction, with a normal stress-reaction concomitant with diabetes insi pidus, and with a lower reactivity to surgical intervention. The treatment algorithms were appropriately amended (the preventive component was added) with due respect to the above circumstances. PMID- 14524026 TI - [Successful therapy of the patient after multiple gunshot wounds with two injured heart chambers and massive blood loss in the early postoperative period]. PMID- 14524025 TI - [Intracellular disorders in myocardium of dogs during cardiogenic shock and possibilities of their correction]. PMID- 14524027 TI - [Acute multiple organ dysfunction syndrome in peritonitis]. AB - The laws of logic and logical research methods were made use of to assess the already published definitions of "polyorgan insufficiency" and "systemic inflammatory response". Many of them were proven to be far from being perfect; hence, the authors' independent understanding of such notions was suggested. PMID- 14524029 TI - [Priorities in anaesthesiology]. AB - On the basis of an analysis of published data, the authors draw the attention of medical circles to generally- and little-known facts of using a variety of general and local anesthesia types by Russian doctors. It is pointed out that not the very first application based on a calendar date of using a method but the first contribution made into the medical science and supported by efforts promoting such method should be regarded as genuine priority. Prominent medical figures are listed, i.e. N.I. Pirogov, S.K. Klikovich, N.P. Kravkov, S.P. Fyodorov, V.K. Anrep, I.N. Katsaurov, A.V. Vishnevsky, whose priority in certain anesthesia variations, as the authors believe it, is indisputable. PMID- 14524028 TI - [Volumetric monitoring based on transpulmonary thermodilution in anesthesiology and intensive care]. AB - The review elucidates the technique of transpulmonary thermodilution in volumetric monitoring of critically ill patients. The experimental and clinical studies, now underway, are indicative of a great value of the discussed method for practical medicine. The key parameters of volumetric monitoring comprise Intrathoracic Blood Volume and Extravascular Lung Water as well as a wide range of other derived variables and indices. The approach is highly effective in the mechanical ventilation and inotope/vasopressor support of patients with a high risk of non-cardiogenic or cardiogenic pulmonary edema. The evaluation of volumetric parameters and of invasive monitoring of hemodynamics provide for an excellent guidance in fluid resuscitation in an overwhelming majority of ICU patients. The PiCCO technology of continuous cardiac output (CO) measurements or, the so-called "pulse contour CO" is also under discussion. PMID- 14524030 TI - [The Military-Medical Academy contribution to the establishment and development of anesthesiology and resuscitation in Russia (to 45th anniversary of the chair for anesthesiology and resuscitation of the Military-Medical Academy)]. AB - Contributions made by some researchers of the Military-Medical Academy (former Imperial Medical-Surgical Academy) made to establishing and promoting the home anesthesiology and intensive care are historically revised in the paper. In particular, the related activity of N.I. Pirogov, S.K. Klikovich, S.P. Kolomnin, S.N. Delitsin, P.A. Kupriyanov and of other widely-known public figures is shortly described. PMID- 14524031 TI - [The value of platelet function tests]. AB - Platelet function tests are used to detect patients with abnormal platelet function, which may be inborn or acquired or to detect increased platelet activation which may be accompanied by an increased risk of thrombosis. Platelet function tests are also used to monitor platelet function inhibitors as aspirin, clopidogrel or platelet membrane glycoprotein IIb/IIIa-inhibitors. Incorrect blood sampling is a major source of error in measuring platelet function. Global tests besides platelet count and bleeding time are thrombelastography, the platelet function analyzer (PFA) and possibly in the future the new Impact system. Specific tests measure platelet spreading and adhesion to defined surfaces. In a series of methods platelets are counted before and after passage of a filter. Some of these tests are partially standardized. The most frequently measured platelet function is aggregation induced by ADP, collagen or other substances as first described by Gustav Born. Some newer methods to perform aggregometry are described. Platelet activation can be detected by measuring spontaneous aggregation as in the PAT-test. Prospective trials with this test have shown that enhanced spontaneous aggregation is a risk factor for new vascular occlusions in diabetics and for myocardial infarctions in healthy individuals. The Wu and Hoak-test and the measurement of released platelet factor 4 or betaglobulin are of limited value. Flow cytometric methods are frequently used to measure platelet activation markers as CD 62 and others. Platelet induced thrombin generation is an interesting function to measure drug effects. None of the presently available platelet function tests is well standardized, so there is much room for improvement. PMID- 14524032 TI - [Pathogenetic significance of methicillin resistant Staphylococcus aureus (MRSA) in chronic wounds]. AB - The colonisation and infection with methicillin-resistant staphylococcus aureus (MRSA) represent an interdisciplinary problem especially in the treatment of chronic wounds. To prevent medical wound-healing centres of becoming vectors for further epidemics it makes sense to take regular bacterial smears of all patients with chronic wounds to identify, isolate and treat patients with MRSA colonisation or infection. Especially infections should only be treated after microbiological diagnostics had been done to allow systemic treatment with antibiotics in accordance with the specific resistances. Otherwise the uncritical use of broad-spectrum- or reserve-antibiotics supports the critical development of further resistances. PMID- 14524033 TI - Association between endothelial dysfunction and major cardiovascular events in peripheral arterial disease. AB - BACKGROUND: Patients with peripheral arterial disease (PAD) are characterized by a high mortality for cardiovascular events. An impairment of endothelial function, expressed as brachial-artery flow-mediated vasodilation (FMV), has been described in PAD patients. Aim of this study was to investigate the association between FMV and cardiovascular events in patients with PAD. PATIENTS AND METHODS: Thirty-eight patients with intermittent claudication (71% men, mean age 71 years) were divided into two groups according to the presence or absence of previous major cardiovascular events (myocardial infarction or stroke). RESULTS: Brachial FMV was significantly lower in patients with a history of myocardial infarction or stroke (n = 16) than in patients without cardiovascular events (3.2 +/- 3.6% vs. 5.7 +/- 3.6%; p = 0.042). In the group with cardiovascular events there was a significantly higher proportion of subjects in the lower FMV tertile (56% vs. 18%), and a lower proportion of subjects in the upper tertile (25% vs. 41%; chi 2 test, p = 0.047). CONCLUSION: We conclude that FMV of the brachial artery is significantly reduced in PAD patients with a history of stroke and myocardial infarction. These cross-sectional results suggest a potential role of FMV as a marker of major cardiovascular events. PMID- 14524034 TI - Hemostatic and fibrinolytic effects of systemic prostaglandin E1 therapy in patients with peripheral arterial disease. AB - BACKGROUND: The study was designed to evaluate if there is any evidence of a hyperfibrinolytic bleeding-risk under systemic treatment with prostaglandin E1 (PGE1) of patients with peripheral arterial disease (PAD). PATIENTS AND METHODS: The in vivo effect of PGE1 on the fibrinolytic and hemostatic process was tested on 15 patients before and after treatment with Alprostadil for 21 days using D dimers (DD), fibrinogen, prothrombin time (PT), partial thromboplastin time (PTT), antithrombin (AT), ProC-Global, plasminogen, plasminogen activator inhibitor activity (PAI), alpha 2-antiplasmin, coagulation factor XII, basal and activated fibrinolytic capacity (fib. cap.). RESULTS: There was no significant difference in DD, fibrinogen, PT, PTT, AT, ProC-Global, plasminogen, PAI, alpha 2 antiplasmin, coagulation factor XII, basal and activated fibrinolytic capacity observed after the treatment. CONCLUSION: Summarizing this study there is no hyperfibrinolytic bleeding-risk after the systemic therapy with Alprostadil to be expected. PMID- 14524035 TI - Surgical treatment of vasculo-Behcet's disease. A review of patients with concomitant multiple aneurysms and venous lesions. AB - BACKGROUND: Vascular complications can be seen in patients with Behcet's disease. Arterial and venous complications may be found separately or concomitantly in patients. PATIENTS AND METHODS: Out of 29 patients with vasculo-Behcet's disease 7 patients with multiple aneurysms and venous lesions were documented over a period of 20 years. All patients were male, ranging in age from 24 to 52 years. The mean duration of the disease was 6 +/- 2 years. The aneurysms were found in the following locations: one pulmonary artery, two abdominal aorta, four iliac, five femoral, and two popliteal artery. Both aneurysmal and occlusive lesions were present in three patients. In the venous lesions associated with the aneurysms there were three deep and three superficial venous thrombosis. Two patients had caval involvement-superior and inferior vena caval syndromes. RESULTS: We performed seven interposition grafting by polytetrafloroethylene, one Y-grafting, one aneurysmorrhaphy, one lobectomy. Re-anastomosis was performed in two patients who had anastomotic aneurysms and graft occlusion without disabling ischemia. Venous pathologies were treated by medical therapy. The patients were followed up between 1 to 8 years. One of the patients with iliac artery aneurysm died due to gastrointestinal bleeding 15 months after the operation. CONCLUSIONS: In conclusion, when an aneurysm has been found in a patient with Behcet's disease, the patient should be scanned for possible multiple aneurysms and venous lesions since they might be found together. Surgical treatment, when feasible, should be performed in cases with Behcet aneurysms because of a high risk of rupture. However, the possibility of an anastomotic aneurysm developing after surgery should also be kept in mind. PMID- 14524036 TI - Vacuum-sealed mesh graft transplantation in chronic cutaneous ulcers of the lower leg. AB - BACKGROUND: The covering of defects caused by chronic ulcers on limbs affected by peripheral arterial disease or chronic venous insufficiency is often difficult due to extensive secretion and edema, while chronic bacterial contamination of the wound bed further compromises the conditions for successful healing. PATIENTS AND METHODS: Vacuum-sealed dressing (VSD) offers the option of a closed dressing system for moist wound care that assures firm contact with the wound surface and protection against contamination with nosocomial microbes and decontamination of existing bacteria by means of constant drainage of secretion independent of gravity. VSD is particularly useful in difficult wounds featuring extensive secretion and unfavorable localization and offers many advantages over conventional dressing techniques in terms of improved healing of skin transplants. A total of 35 patients with chronic leg ulcers were treated with vacuum-sealed mesh graft transplantation. RESULTS: Complete healing of the mesh graft transplant was observed in 20 patients (57%). Twelve patients (34%) experienced partial healing (75-90%) of the transplant, while three patients exhibited less than 75% healing of the graft and therefore required a second mesh graft transplantation. CONCLUSIONS: VSD is a simple, quick and inexpensive technique that promotes excellent healing of skin transplants. It is particularly useful in difficult wounds with extensive secretion and/or contamination or infection. PMID- 14524037 TI - Proteus syndrome. AB - A 34-year-old male patient was referred with a recalcitrant leg ulcer overlying an extensive vascular malformation, which had led several times to septic soft tissue infections. During his infancy he had been diagnosed to have Klippel Trenaunay syndrome. Clinical examination revealed asymmetric hypertrophy of the lower extremities, an extensive portwine stain on the more severely affected left limb as well as prominent venous varicosities of both legs. Hands and feet showed striking cerebriform palmoplantar hypertrophy, and macrodactily with syndactily of several fingers. All toes had been amputated in early childhood due to extreme overgrowth and currently the patient walked on his forefeet in a prominent pes equinus deformity. Further symptoms consisted in several lipomas at both arms, another portwine stain at the left hemithorax and a single cafe-au-lait spot at the left scapula. Angio-magnetic resonance imaging scans of both legs showed an extensive venous-lymphatic vascular malformation involving the whole subcutis and infiltrating the muscle. The chronic wound was interpreted as venous stasis ulceration. Local percutaneous sclerotherapy of the dilated veins underneath the ulcer was discussed, but considered to carry a relevant risk of skin necrosis with consecutive progression of the wound. A conventional split-skin graft led to complete wound healing. Since, the patient consequently wears custom-made compression stockings and remained free from recurrences. The syndromatic constellation of palmoplantar overgrowth, multiple lipomas, giant fingers and toes, limb overgrowth, venous-lymphatic malformation and a cafe-au-lait spot led to the diagnosis of Proteus syndrome. The possible aetiology, clinical manifestations, differential diagnosis and management of this rare disorder are discussed. PMID- 14524038 TI - Venous abnormalities encountered in abdominal aortic aneurysm repair. AB - The coexistence of abdominal aortic aneurysm (AAA) and venous abnormalities is unusual. In this article two cases of AAA with concomitant abnormalities of a retroaortic left renal vein and left-sided inferior vena cava are presented. PMID- 14524039 TI - An unusual complication of open aortic surgery: hemorrhage due to the iatrogenic rupture of a renal cyst. AB - The authors describe a clinical case of a retroperitoneal hemorrhage due to rupture of a voluminous renal cyst during a conventional open aortic surgery. Intraoperative trauma is to be considered the cause of bleeding. In most cases the clinical evolution of the patient is benign and conservative treatment is sufficient. Considering the incidence of renal cysts in patients ungergoing conventional aortic surgery, we want to emphasize this possible complication, that has only rarely been dealt with in the literature. We recommend evaluation of such patients in view of preoperative drainage of large cyst including injection of sclerosing agents. PMID- 14524040 TI - Endotension leading to aneurysm sac rupture following open repair of abdominal aortic aneurysm. AB - Endotension leading to enlargement of the aneurysm sac following the endoluminal grafting is still handled as an exclusive phenomenon of the endovascular aneurysm repair (EVAR). We report on a case with aneurysm sac enlargement caused by endotension leading to aneurysm rupture after conventional, open aneurysm repair, a so far not described complication. In a 74-year-old patient, following open surgical standard resection and reconstruction of an abdominal aortic aneurysm, a routinely performed abdominal ultra-sonography demonstrated a slowly growing enlargement of the peri-prosthetic aneurysm sac without endoleak. During the pre operative work-up of the cardiac and pulmonary risk profiles, he complained of abdominal pain and back pain. Control CT revealed contrast inside the aneurysm sac as well as in the right-sided retroperitoneum. At the emergency operation a retroperitoneal haematoma was noticed. Opening the ballooned aneurysm sac, a fresh haematoma was also found. Lifting up the prosthesis, back bleeding at the dorsal circumference of the proximal anastomosis was confirmed due to a 2 cm long disruption of the anastomosis. In patients who present with abdominal or back pain after conventional surgery of an abdominal aortic aneurysm, a contrast CT should be performed to exclude an endoleak as well as other pathologies. The enlargement of the aneurysm sac without endoleak could be interpreted as endotension, with the consequence of urgent re-operation to prevent rupture. PMID- 14524041 TI - Fibrinogen adsorption--a new treatment option for venous leg ulcers? AB - The initial element in the causation of venous ulceration is a disturbance of venous blood flow that leads to an increase in venous pressure. Eventually, however, it is the microcirculatory consequences of venous hypertension that lead to trophic skin changes and finally to ulceration. A reduction in blood viscosity results in an improvement at the microcirculatory level. The elimination of fibrinogen from plasma improves blood viscosity. This case report concerns a 75 year-old woman with venous ulcers of both legs (left lower leg: deep ulceration with a surface area of 3 x 5 cm; right lower leg: superficial, confluent ulceration with a total surface area of 5 x 10 cm). The patient underwent 20 sessions of fibrinogen adsorption, while simultaneously continuing with a regimen of conservative measures (activated charcoal cloth dressing with silver, calcium alginate dressings and short-stretch compression bandages). Following binding to a peptide (Gly-Pro-Arg-Pro-Lys), fibrinogen and fibrin were specifically removed from the patient's plasma: her fibrinogen concentration was lowered from an original mean level of 310 mg/dl (SD +/- 104 mg/dl) to 136 mg/dl (SD +/- 54 mg/dl), and there was no return to the baseline concentration by the time of the next fibrinogen adsorption session. In response to this treatment the patient's ulcers healed rapidly within 9 weeks. Dizziness and hematomas at the vascular access sites in both antecubital fossae were reported as adverse effects. A fall in hematocrit was also noted (before treatment 37% +/- 1%; after treatment 35% +/ 2%). This may have been caused by hemodilution due to the procedure and to cell losses during blood-plasma separation, a phenomenon that is known to occur during apheresis. This case report suggests that fibrinogen adsorption is low in adverse effects and is a useful addition to the range of treatments available for ulcers of venous etiology. PMID- 14524042 TI - Aneurysms of radial arteries and severe anemia. PMID- 14524043 TI - [Peripheral arterial occlusive disease (PAVK)]. PMID- 14524044 TI - [Conservative treatment of acute embolism occlusions of the carotid bifurcation]. PMID- 14524045 TI - The dual imperative in refugee research: some methodological and ethical considerations in social science research on forced migration. AB - Social scientists doing fieldwork in humanitarian situations often face a dual imperative: research should be both academically sound and policy relevant. We argue that much of the current research on forced migration is based on unsound methodology, and that the data and subsequent policy conclusions are often flawed or ethically suspect. This paper identifies some key methodological and ethical problems confronting social scientists studying forced migrants or their hosts. These problems include non-representativeness and bias, issues arising from working in unfamiliar contexts including translation and the use of local researchers, and ethical dilemmas including security and confidentiality issues and whether researchers are doing enough to 'do no harm'. The second part of the paper reviews the authors' own efforts to conduct research on urban refugees in Johannesburg. It concludes that while there is no single 'best practice' for refugee research, refugee studies would advance its academic and policy relevance by more seriously considering methodological and ethical concerns. PMID- 14524046 TI - Early socio-political and environmental consequences of the Prestige oil spill in Galicia. AB - The controversial form in which the oil industry is run has once more caused a huge disaster--this one affecting the Galician coastal environment and economy. Oil-spill clean-up operations have been managed in Europe with some success but with considerable economic, environmental and social costs. The oil industry often avoids fully or even partially compensating those affected. The lack of both political will and political power has let the culprit (the oil industry) off the hook. This paper considers the spill of the Prestige to assess whether the balance of power between affected people and the oil industry can be changed. The paper examines the growing awareness of environmental issues among ordinary people in Spain, through the massive involvement of volunteers concerned with the damage done to the environment and to the livelihoods of fishing communities in Galicia. To understand these growing public concerns and the strength of opinion, the paper examines the details of the decisions taken by the central Spanish and local governments and the way these have informed the clean-up operations, the character of the oil companies involved and the feeling of impotence in the face of such disasters. The conclusion here is that the operations of the oil industry should be tightly regulated through EU legislation, and that this can come about as a result of organised political pressure from those affected by the oil spill, from the mass of volunteers, as well as from public opinion at large. PMID- 14524047 TI - Constructing vulnerability: the historical, natural and social generation of flooding in metropolitan Manila. AB - Flooding is not a recent hazard in the Philippines but one that has occurred throughout the recorded history of the archipelago. On the one hand, it is related to a wider global ecological crisis to do with climatic change and rising sea levels but on the other hand, it is also the effect of more localised human activities. A whole range of socio-economic factors such as land use practices, living standards and policy responses are increasingly influencing the frequency of natural hazards such as floods and the corresponding occurrence of disasters. In particular, the reason why flooding has come to pose such a pervasive risk to the residents of metropolitan Manila has its basis in a complex mix of inter relating factors that emphasise how the nature of vulnerability is constructed through the lack of mutuality between environment and human activity over time. This paper examines three aspects of this flooding: first, the importance of an historical approach in understanding how hazards are generated; second, the degree of interplay between environment and society in creating risk; and third, the manner in which vulnerability is a complex construction. PMID- 14524048 TI - The Afghan experiment: the Afghan Support Group, Principled Common Programming and the Strategic Framework. AB - From 1997 to 2001, the international community put in place unique mechanisms to address the challenges to providing humanitarian assistance in Taliban-run Afghanistan. The Afghan Support Group (ASG), Principled Common Programming (PCP) and the Strategic Framework (SF) constituted a precedent-setting experiment that consumed thousands of hours in implementation in donor capitals and in the field. This article, written from the perspective of the leading donor of humanitarian assistance to Afghanistan, assesses the extent to which this Afghan experiment succeeded in increasing coherence on assistance policy issues, improving efficiency in assistance programmes and added synergy between assistance and peace efforts; identifies factors that limited further achievements; suggests how the mechanisms could have been improved; and analyses whether the overall effort politicised humanitarian assistance. The article concludes that the record of achievement was sufficiently promising that the ASG/PCP/SF experiment should be considered for application in other complex humanitarian emergencies. PMID- 14524049 TI - Humanitarian Daily Rations: the need for evaluation and guidelines. AB - Over 13 million Humanitarian Daily Rations (HDRs) have been distributed by NGOs, international organisations and the US military since 1993. In that time, not a single technical end-user study of HDRs has been published. Nor have practical guidelines been established for HDR distribution or use. This paper seeks to establish a baseline of knowledge concerning HDRs, surveying their history, composition and distribution. Criticisms of the ration are reviewed. A comprehensive evaluation is required to understand and improve HDR distribution, end-use, perceptions, design and nutritional contents. HDRs must also be added to technical guidelines on planning nutritionally adequate rations. PMID- 14524050 TI - [Quality assurance of rheumatologic patient education]. AB - Since 1989 patient education programmes for patients with rheumatoid arthritis, ankylosing spondylitis and other spondylarthropathies, systemic lupus erythematosus, vasculitis, fibromyalgia, and juvenile chronic arthritis and their parents have been developed by an interdisciplinary team of the German Society of Rheumatology (Deutsche Gesellschaft fur Rheumatologie). Up to date, about 500 people were trained to be group leaders or specialised trainers with an associated train-the-trainer program. In 1999, the Society discharged preliminary guidelines for patient education in rheumatology. Prospective randomised studies demonstrated that patient education led both to an improvement of knowledge, self efficacy and self-help activities and to an improvement of arthritis-related helplessness and pain, and a reduction of both temporary and permanent disability. PMID- 14524051 TI - [An example of internet-based quality assurance in rheumatology]. AB - In Germany, patient care and medical progress have become increasingly influenced by quality assurance and quality management and also by a rapid increase in knowledge and information. Thus, modern information management achieves high relevance in medicine, too. The internet provides a fascinating medium with its open architecture and interaction possibilities offering new, trailblazing perspectives. This article shows examples of the current use of the internet for quality assurance at the RheumaNet, which functions as a presentation platform for the competence network for systemic inflammatory rheumatic diseases ('KN Rheuma') in Germany. The RheumaNet has already contributed to achieving quality assurance by supplying extensive, valuable and reliable scientific information. Other innovative interactive and quality assuring applications have been developed, tested and implemented in the system. Future applications will deal with standardised online documentation modules, and aids for patient self management are on their way. PMID- 14524053 TI - [Quality management of treatment of rheumatoid arthritis in a rheumatological setting]. AB - Rheumatoid arthritis is a chronic inflammatory rheumatic disease affecting about 1% of the general population world-wide. It is characterised by severe pain and a reduction of functional capacity leading to a reduced quality of life. The initiation of an early, effective, continuous and long-term treatment is essential for preventing or delaying progression of disease as long as possible. The implementation of a comprehensive and structured quality management program including both general practitioners and specialists in rheumatology will help to support structural, procedural and outcome quality based on special indices that can be used for benchmarking. The Swiss Clinical Quality Management (SCQM) and the regional model project of the Regional Co-operative Rheumatology Centre in Hanover, Germany (Regionales Kooperatives Rheumazentrum Hannover e.V.) are two examples for total quality management (TQM) of inflammatory rheumatic diseases. PMID- 14524052 TI - [Rheumatology education in quality circles of office-based general practitioners: A novel andragogic concept for continuing medical education]. AB - Continuing medical education is essential to improve the quality of health care delivery. The efficacy of postgraduate medical education generally improves when participants feel that their specific needs are met by a particular educational activity. Audit circles of general practitioners allow for a discussion of relevant issues. The present paper describes a novel concept for delivering knowledge and skills in arthritis and rheumatic conditions to general practitioners. A well-designed paper case was used for discussion in established audit circles of general practitioners, and a specially trained expert rheumatologist participated as an external expert to answer all those questions that could not be solved by the other participants. The participation of such as an expert-on-demand was found extremely helpful, as an evaluation during pilot sessions in 6 pre-existing audit circles of general practitioners revealed. PMID- 14524054 TI - [The European Regional Cooperative Centers for rheumatic diseases: a nation-wide network of interdisciplinary rheumatic care]]. AB - The organisation of regional co-operative centres for rheumatic diseases was supported by a grant from the German Ministry of Health from 1992 to 1998, which aimed to improve the management of patients with chronic rheumatic diseases. After establishing the centres formal structures the major task was to improve quality of care by co-ordination and co-operation of all the institutions and health care providers involved in the management of rheumatic patients. For all these activities to be continued the regional co-operative centres for rheumatic diseases were integrated in the German Society of Rheumatology as one of its working groups in 1996. The major objective is the quality management of rheumatic care including a uniform patient documentation (the so-called core documentation), a thesaurus of diagnoses and the nation-wide use of a patient passport. Most importantly, guidelines for diagnosis and therapy were developed and especially adapted for use by patients and primary care physicians. Five project working groups (guidelines for diagnosis and therapy, documentation, quality management, paediatric rheumatology, physical medicine and rehabilitation) were established to work on future initiatives. Quality management is further supported and intensified by annual scientific workshops. In addition, the regional co-operative centres for rheumatic diseases are engaged in research and an important partner of the German Competence Network Rheumatic Diseases. Based on the established structures the horizontal and vertical network should be further involved in general medical care and should intensify the interaction with clinical and basic research. PMID- 14524055 TI - [The National Database of the Regional Collaborative Rheumatic Centers as a tool for clinical epidemiology and quality assessment in rheumatology]. AB - The national database of the German Collaborative Arthritis Centres is a well established tool for the observation and assessment of health care delivery to patients with rheumatic diseases in Germany. The discussion of variations in treatment practices contributes to the internal quality assessment in the participating arthritis centres. This documentation has shown deficits in primary health care including late referral to a rheumatologist, undertreatment with disease-modifying drugs and complementary therapies. In rheumatology, there is a trend towards early, intensive medical treatment including combination therapy. The frequency and length of inpatient hospital and rehabilitation treatments is decreasing, while active physiotherapy in outpatient care has been increased. Specific deficits have been identified concerning the provision of occupational therapy services and patient education. PMID- 14524057 TI - [Experiences from practical guideline development--proposal of a more standardized guideline development process]. AB - In this article, the authors of the ,,General Practitioners' Guideline for the Diagnosis and Treatment of Urinary Incontinence" describe the logical and procedural steps involved in the development process of this evidence-based GPs' guideline for the management of urinary incontinence. Specifically, the following factors have been identified as crucial for a successful primary care guideline development process: The work group (authors) should comply in size with current recommendations for successful project work teams. It is recommended that authors should belong to opposing ends of the spectrum of practical versus theoretical medicine. We argue that the integration of both practical experience and theoretical knowledge/research experience be combined in order to collate guideline recommendations that are firmly grounded in scientific evidence and both applicable and acceptable in everyday practice. Proper work on a guideline development project should only start after ensuring that the well-known theoretical requirements for sensible guidelines are met, i.e. the problem in our special focus should be widespread or of considerable economic/public health importance, be amenable to proper medical intervention, and some quality issue regarding the current handling of this problem should have been demonstrated. Subsequently, a clear logical analysis of the problem's inherent functional/logical/practical subdivisions has to be conducted. Each author is then allocated his or her fair share of the work. The next step is considered to be extremely important and consists of discovering one's "illusive correlation", i.e. inherent ideas one has on the subject without being aware of one's own bias. The proposed "trick" now consists of making the implicit bias an explicit fact in the further guideline development process. The remaining work steps build on the integration of one's preformed ideas with current, published opinion from standard textbooks, as well as existing guidelines and non-guideline recommendations on the subject; plus taking into account the practical experience gathered from interviews and "on-location-visits". This integration process is performed within a structured framework to facilitate objectivity and transparency of the entire guideline. PMID- 14524056 TI - [Physicians and the internet--a cross-sectional study against the background of guideline implementation]. AB - The internet is an innovative medium for the implementation of current recommendations for diagnosis and therapy, e.g. by means of guidelines. In this context, evaluation of the internet is very important because traditional ways of implementation have been proved less effective. Therefore, we investigated the frequency of online access and individual utilisation of the internet among 13,547 family practitioners, internists and general practitioners using a questionnaire procedure. Furthermore, we asked for a personal grading of its current relevance in their daily practice. Out of 2,786 responders (20.6% response rate) 79% reported personal online access. 40% had online computer access in their office and 71% at home. 45% of the internists had online access in their office compared to 34% of general practitioners. Almost all physicians under the age of 40 years (94%) had personal online access in comparison to only 49% of those over 60 years. The average daily duration of internet usage was up to ten minutes in 61%, and 1.5% use the internet more than one hour per day. 46% of responders believe that the internet is an appropriate source for professional education, whereas it has little professional relevance for 38%. We conclude that the internet appears to be a useful medium for the implementation of guidelines. However, in subgroups with the most urgent need for current medical information the internet seems to be unsuitable as an implementation tool. PMID- 14524058 TI - [Quality of antithrombotic therapy in patients with chronic atrial fibrillation: the AFib Trial]. AB - BACKGROUND: Large randomised studies have definitely shown that oral anticoagulation effectively reduces thromboembolic complications, e.g. stroke, in patients with chronic non-valvular atrial fibrillation (AFib). However, less than 50% of the eligible AFib patients receive anticoagulation, although their risk of thromboembolic disease is increased 5-7 fold as compared to individuals who have a regular sinus rhythm. This is the reason why undertreatment needs to be evaluated and addressed. In Germany, data on the antithrombotic therapy of these patients are sparse, national guidelines are lacking. AIM OF THE STUDY: This prospective cohort study is the first one to evaluate the quality of antithrombotic prevention in two groups of AFib outpatients in two different, socio-demographically comparable regions of Germany (Sudbaden and Sudwurttemberg) over a period of several years. Quality of care is defined as the percentage of AFib patients in a certain region treated according to international guidelines. If the percentage in both cohorts does not exceed 80%, guidelines are to be developed and implemented in Sudwurttemberg on consideration of the specific treatment conditions in this region; their applicability is to be evaluated later. METHODS: Office-based specialists of internal and general medicine are currently recruiting 200 AFib patients in each study region for documentation of their clinical data. Besides the quality of antithrombotic chemoprevention, relevant problems emerging in antithrombotic therapy in an ambulant setting will be identified. In the area of intervention (Sudwurttemberg) experts and office based physicians will exclusively develop evidence-based guidelines and disseminate them among doctors. Six months after the implementation of these guidelines in Sudwurttemberg, their influence on prescribing patterns will be determined by comparing the proportion of anticoagulated AFib patients in Sudwurttemberg to that of the control cohort (Sudbaden). In addition, secondary outcomes of interest include deaths, days of hospitalisation, and incidence of stroke or bleeding episodes for different antithrombotic treatments. PMID- 14524059 TI - [Joint diseases in X-ray photography]. PMID- 14524060 TI - [Foot abnormalities -- stress fractures -- wrong footwear. Jogger with foot disease ]. PMID- 14524061 TI - [For each diabetic the right program. Insulin therapy according to the physiology model]. PMID- 14524062 TI - [Colleagues' tips against recurrent cystitis. Cranberry juice instead of antibiotics]. PMID- 14524063 TI - [Life -- a unique risk factor. Is the man only a conglomerate of diseases?]. PMID- 14524064 TI - [Smallpox vaccination. The USA wants our vaccines]. PMID- 14524065 TI - [Multiple chemical sensitivity. Environment victim or imaginary patient?]. PMID- 14524066 TI - [Diet therapy in hypertension, disordered lipid metabolism, diabetes and gout. A dietary plan for all cases]. PMID- 14524067 TI - [Diet counseling for hypertensive patients. What measures make sense?]. AB - The leading principles of diet and lifestyle in the treatment of hypertension are identical to those applicable to elevated LDL cholesterol. The following measures or recommendations apply: fewer saturated fatty acids, weight reduction, more exercise, more fruit and vegetables, alcohol--if at all--strictly in moderation, no smoking. Patients with hypertriglyceridemia should give greater preference to vegetables rather than fruits with their higher content of fructose. The effect on the blood pressure is greater the more of these principles are applied. As in the case of blood pressure-lowering drugs, the response of the individual patient to dietary measures also varies considerably. Nevertheless, lifestyle measures should definitely not be dismissed. Rather, the individual principles should be investigated for their efficacy. PMID- 14524068 TI - [Diet in disordered lipid metabolism. A culinary balance act]. AB - When LDL cholesterol is elevated, HDL cholesterol is low or triglycerides are raised, dietary changes form the basis of treatment. Such changes are most important in the case of hypertriglyceridemia. Some 3 to 4 hours after a meal, triglycerides increase to an extent determined by the composition of the meal. Hypertriglyceridemia cannot be successfully treated unless alcohol is banished and rapidly assimilatable carbohydrates are restricted. In patients with elevated LDL cholesterol, a change in eating habits can have an appreciable effect (on average 10-15%). Since this measure can save the use or reduce the dose of medications, its value is obvious, and it must not be neglected. The measures aimed at elevating HDL cholesterol have only a moderate effect, so that more importance should be attached to lowering the LDL fraction. PMID- 14524069 TI - [Just as important as blood sugar control by drugs. A healthy cuisine for diabetics]. AB - A modern optimal diet for diabetics offers more than simple control of blood sugar. Rather, it also aims to minimize the additional cardiovascular risk factors that are often present: hypertension and lipometabolic disorders as well as hyperuricemia. An improvement in the prognosis in particular of patients with a full-blown metabolic syndrome can be achieved only by comprehensively treating the various disorders associated with the disease. A specific "diabetic diet" is unknown. For the most part, recommendations focus on an energy-reduced, moderate fat, high bulk, mixed diet. An additional aim is reduction of overweight with a BMI target of < 25 kg/m2. Since the eating habits of German diabetics are currently far from complying with these recommendations, greater engagement is required not only from the patient, but also from the care-providing physician. The latter should ensure that the patient receives qualified instruction, which can appreciably improve the patient's motivation and cooperation. PMID- 14524071 TI - [Suicide prevention by the general practitioner]. AB - Although suicides in Germany are on the decrease, for the care-providing physician a suicidal patient must always be considered an emergency. An elevated risk is to be seen above all in patients with psychological/mental problems, in particular those suffering from depression, but also those experiencing a crisis associated with ageing, physical illnesses or intolerable social situations. A range of factors may influence suicidal behavior, such as the specific situation of the patient or the loss of protective factors. When his/her coping strategies or outside help then fails, a triggering event may prompt suicide. As a rule, the suicide-endangered patient goes through three stages--a period of weighing up, ambivalence and decision-taking. For the care-providing physician it is of particular importance to recognize the ambivalence stage in good time, for it is here that the patient gives expression to thoughts of suicide and the physician has an opportunity to intervene. Major preventive measures are the build-up of a doctor-patient relationship and a readiness to "talk things through". Further crisis management includes, for example, the use of psychopharmaceuticals or the initiation of psychotherapy. PMID- 14524070 TI - [Avoid purine-rich foods, drink a lot, reduce weight. The most important recipes against hyperuricemia]. AB - In 99% of the cases, the underlying cause of gout is an inborn disorder of uric acid excretion. In addition to acute arthritis, clinical consequences include tophi, chronic destruction of joints, and ulcers. The patient is endangered by the frequent renal involvement, with hypertension and renal insufficiency. Furthered by such exogenous factors as overweight, high-calorie, purine-rich foods and immoderate consumption of alcohol, hyperuricemia may develop. Basic treatment comprises dietary measures and, up to a serum uric acid concentration of 8.5 mg/dl, should be the sole measure required. The aim is to achieve a permanent lowering of uric acid concentrations in the blood to < 6.5 mg/dl. This may be achieved by reduction of overweight (including regular exercise), an energy-adapted diet with only moderate amounts of fat, restriction--or, better, banishment--of alcohol, an ovolactovegetarian diet low in purine (care must be taken with high-purine vegetablesl), moderate intake of readily assimilated carbohydrates, and adequate amounts of calorie-free liquids. PMID- 14524072 TI - [Effective protest against reform plans. Physician's sourness toward woman politician]. PMID- 14524073 TI - [Insurance and health care reform create problems. EBM decision is indeed postponed again]. PMID- 14524074 TI - [Steroid therapy with small risk. Intestinal inflammations to be treated topically]. PMID- 14524075 TI - [American hypertension guidelines discussed. Are diuretics really the best first line therapy?]. PMID- 14524076 TI - [Common airway ailments. What does homeopathy bring?]. PMID- 14524077 TI - [Asthma therapy. A new steroid will be active first in the lung]. PMID- 14524078 TI - [Strong and sustained-acting triptan. Therewith migraine does not return soon]. PMID- 14524079 TI - [Painless blood sugar self-monitoring. Arm instead of finger]. PMID- 14524080 TI - [Erectile dysfunction to be taken seriously. Otherwise, the psyche becomes limp, too]. PMID- 14524081 TI - [Osteoporosis therapy compared. More solid bones with bisphosphonates]. PMID- 14524082 TI - [Diuretics in heart failure. Fewer electrolyte disorders -- better prognosis]. PMID- 14524083 TI - [Selective COX 2 inhibitor rofecoxib. To "defy" the pain]. PMID- 14524084 TI - [Meta-analysis on acute pain. Rofecoxib helped three quarters of patients]. PMID- 14524085 TI - [Hirsutism]. PMID- 14524086 TI - [Magnification endoscopy zoom endoscopy]. PMID- 14524088 TI - [Some specific features of drug resistance of Mycobacteria tuberculosis in patients with acutely progressive destructive forms of pulmonary tuberculosis]. AB - A clear correlation has been found between the drug resistance of Mycobacterium tuberculosis (MBT) and the degree of immunodeficiency in 127 patients with acute destructive tuberculosis. The findings indicate that it is possible to use significant immunodeficiency as an early marker of the multidrug resistance of MBT and as a predictor of cheesy pneumonia. The fact that there was X-ray involution in one lung and concomitantly progression in the other suggests the presence of MBT strains showing different drug resistance. The efficiency of treatment for acute tuberculosis has been ascertained to be determined by MBT drug resistance. PMID- 14524087 TI - [Continuation 47. Dyspnea is the main symptom. It's up to you to clarify it!]. PMID- 14524089 TI - [Tuberculin sensitivity in infants and preschool children with pulmonary tuberculosis]. AB - The tuberculin sensitivity of the skin was studied in children with tuberculosis, whose age was under 7 years, in relation to the age, the form of tuberculosis, the phases of a process, its severity, the presence of complications and concomitant disease. It was established that in infants the rate of negative reactions was 13.2 times higher and that of hyperergic reactions was 5.8 times less than those in children aged 3 to 7 years. Negative reactions were most common in the complicated and severe forms of tuberculosis and in comorbidity. Hyperergic reactions were more frequently encountered in ocular tuberculosis (44.5%), helminthic invasion (30.4%), and pulmonary tuberculosis first detected in the phase of resolution (32.4%). PMID- 14524090 TI - [Impact of cytomegalovirus infection on the course of tuberculosis in children and adolescents]. AB - Sixty-five children and adolescents with respiratory tuberculosis were examined to determine the proportion of cytomegalovirus infection among patients with tuberculosis and its impact on the occurrence and course of different forms of tuberculosis. The number of children and adolescents infected with cytomegalovirus among the patients with respiratory tuberculosis (66.2%) was found to be three times more than that among those not infected with tuberculosis (21.0). The absolute majority (91.3%) of the examinees with the complicated course of a tuberculous process were carriers of cytomegalovirus. They were found to have both antigens and antibodies to cytomegalovirus 9 times more frequently than patients with the smooth course of tuberculosis, which is indicative of the activity of cytomegalovirus infection. The findings suggest that cytomegalovirus infection has an impact on the occurrence and pattern of a tuberculous process. PMID- 14524091 TI - [Comparative analysis of a new modification of segmental tuberculin test with the routine Koch test]. AB - A focal reaction plays the leading role in the assessment of segmental tests (ST). The sensitivity of ST is 72.4% and that of the Koch test is 64.5%, their specificity is 94.1 and 80.0%, respectively. A 10% or more increase in adenosine deaminase during peritoneal fluid ST indicates to the exacerbation of genital tuberculosis. Analysis of the cellular composition of peritoneal fluid from the Douglas space is proposed as a criterion for a focal reaction. There are ultrasound symptoms which accompany the exacerbation of an inflammatory process in the genitals in response to ST and which are indicative of a positive focal reaction. The minimum tuberculin load and lack of contraindications allow ST to be introduced into the practical work of tuberculosis facilities. PMID- 14524092 TI - [Functional studies in the prediction of postoperative complications in the presence of lung diseases]. AB - Data on the functional status of the cardiorespiratory system are required to identify patients at risk for postoperative complication in the presence of lung diseases. Very many factors influence the course of an operation and the postoperative period so there is no golden standard or the only parameter for predicting how the postoperative period runs. Patients with normal spirographic values (FEV1, more than 80%??) and without cardiovascular comorbidity are at a slight risk for postoperative complications. These patients do not need to be additionally examined. A less than one-month history of myocardial infarction, instable angina pectoris, decompensated heart failure, severe valvular disease are contraindications to planned surgery. The risk of cardiovascular events is high when the signs of myocardial ischemia occur with low exercise (less than 4 MET). Stress echocardiography, loading tests, and radioisotopic study are used as auxiliary techniques, FEV1, under 60%; ppo-FEV1, and ppo-DC, under 40%; VO2max, under 15 ml/kg/min are the values of a high risk for respiratory complications. PMID- 14524093 TI - [Significance of hematuria in phthisiourology]. AB - The case histories of 299 patients examined at the Urogenital Department, Novosibirsk Research Institute of Tuberculosis, were analyzed to define the etiology of urogenital diseases. The diagnosis of urogenital tuberculosis was established in 112 (37.4%) patients; that of chronic nonspecific pyclonephritis was in 90 (30.1%) patients; chronic nonspecific prostatitis was detected in 49 (16.4%) males; urolithiasis was found in 20 (6.7%) patients. Twelve (4%) and 60 (20.1%) patients were diagnosed as having urinary system cancer and hematuria, respectively. Hematuria most frequently (75%) was indicative of the presence of a tumor. However, in renal tuberculosis, this sign was also revealed in 24.1% of the patients. It should be noted that there is a recent tendency for an increase in the incidence of diseases concurrent with hematuria: the kidney has been found to be concomitantly afflicted with tuberculosis and cancer in 2 patients; tuberculosis is concurrent with nephrolithiasis in 2 other patients. PMID- 14524095 TI - [Lisinopril in the treatment of patients with chronic cor pulmonale]. PMID- 14524094 TI - [Experience in treating patients with tuberculous spondylitis]. AB - Some experience in treating 144 patients with tuberculous spondylitis is presented. The cervical, thoracic, and lumbar parts of the spine were affected in 12, 83, and 49 patients, respectively. Medical and surgical treatment was performed in 82 (57%) and 62 (43%) patients, respectively. The medical treatment included the use of antituberculous drugs, pathogenetic exposures, and an orthopedic regimen. The surgical treatment was a stage of multimodality therapy in 62 patients and it involved the radical elimination of a specific inflammation focus, followed by bone defect replacement and vertebral ceramic implant stabilization. The long-term results were studied in the periods of 2 to 10 years. All the patients were found to have a formed osseous block: exacerbations and recurrences being absent. PMID- 14524096 TI - [The cardiovascular system in patients with bronchial asthma of varying severity]. AB - Seventy patients with bronchial asthma (BA), including 17 with mild asthma, 41 with moderate asthma, and 12 with severe BA, were examined by performing external respiration function test, echocardioscopy, ECG, 24-hour Holter monitoring, lipid metabolic studies. Myocardial ischemia was detected in 47% of the patients with BA: in 56 and 83% with moderate and severe BA, respectively. Silent myocardial ischemia was found in 32% of the patients with moderate BA. In severe BA, there was only painful myocardial ischemia. Diastolic dysfunction of the right ventricle (RV) was the earliest hemodynamic change in BA; RV hypertrophy and dilation and left ventricular diastolic dysfunction were observed in severe BA. PMID- 14524097 TI - [Life quality in patients with sarcoidosis and isolated skin syndrome in chronic dermatoses]. AB - Seventeen patients, including 3 (17.6%) males and 14 (82.4%) females, who had respiratory sarcoidosis with the skin syndrome as erythema nodosum, were examined; their mean age was 41.65 +/- 2.63 years. The histological verification of the diagnosis was 94%. Among 77 patients with chronic dermatoses with the isolated skin syndrome (psoriasis, acne vulgaris, atopic dermatitis, neurodermitis, lichen ruber planus, seborrheal pemphigns, eczema), there were 33 (42.9%) males and 44 (57.1%) females, whose mean age was 36.64 +/- 1.3 years. Comparative analysis of life quality (LQ) in these two groups by using the WHOQOL 100 questionnaire revealed differences in two indices that reflect the level of independence and social relations (p < or = 0.05). In terms of independence, the impact of the disease on LQ was more pronounced in patients with sarcoidosis, its negative effect being in 75% of the criteria rated. Thus, poorer LQ may be stated in patients with skin lesion than in those with the isolated skin syndrome in chronic dermatoses. PMID- 14524098 TI - [The clinical picture and course of sarcoidosis under the conditions of the West Ural]. AB - The specific features of the clinical picture and course of respiratory sarcoidosis (RS) and the efficiency of its treatment were studied in its new cases living in the Perm Region (the West Urals). There was a higher incidence of RS in inhabitants from towns with developed heavy industry, mainly in females. Primary chronic RS was prevalent (89.9%). The therapeutic efficiency was 69.5%. The outcomes of treatment were worse when the disease was untimely detected. Cure rates were higher and slightly lower when corticosteroids as tablets were used in the continuous (87.5%) and intermittent (89.9%) regimens, respectively, in combination with pathogenetic agents and treatments aimed at preventing the development of postsarcoid pneumosclerosis. PMID- 14524099 TI - [Comparison of nitrate reductase and automatic BACTEC MGIT 960 AST techniques for determining the drug sensitivity of mycobacteria tuberculosis]. AB - The accelerated nitrate reductase method (NRM) developed at the Central Research Institute of Tuberculosis versus the automatic assay of drug sensitivity by means of a BACTEC 960 bacteriological analyzer was assessed. NRM was carried out, by using the Lowenstein-Jensen medium for 10 days. It is based on the detection of alive Mycobacteria tuberculosis, by recording their enzymatic activity. The study showed a good agreement of the results obtained by NRM with those obtained on a BACTEC 960 analyzer. Agreements were found for 52 isolates in 47 (90.4%) cases, the results disagreed in the testing of 5 (9.6%) cultures. The results of NRM were identical to those for 21 of the 22 cultures sensitive on a BACTEC 960 device; the coincidence was 95.5%. The sensitivity of NRM ranged from 88.2% (for rifampicin) to 96.3% (for isoniazid) and the specificity did from 96% (for isoniazid) to 100% (for streptomycin, rifampicin, and ethambutol). The positive prognostic value of NRM was 100% (for streptomycin, rifampicin, and ethambutol) and 96.3% (for isoniazid). The negative prognostic value of NRM ranged from 94.6 to 96.8% for individual drugs. The efficiency of NRM (a ratio of the number of correct results to the total number of results) was greater than 0.96, which suggests that this method and the BACTEC MGIT 960 AST technique may be regarded as rather comparable. The testing of NRM versus the automatic BACTEC MGIT 960 AST technique has indicated that the former may be successfully used to determine the sensitivity of Mycobacteria to the critical concentrations of first-line antituberculous agents. PMID- 14524100 TI - [Impaired balance of the lung proteolytic system in patients with pneumoconiosis and chronic dust-induced bronchitis]. AB - The activity of neutrophilic elastase and the level of its major inhibitor--an alpha 1-inhibitor of proteinases were studied to evaluate the protease antiprotease system in the bronchoalveolar lavage fluid (BALF) of patients with chronic dust-induced bronchitis (CDB) and pneumoconiosis (PC). It was found that CDB, as compared with PC, was characterized by a higher elastase activity in the BALF (70%) and that there were a larger number of patients with elastase activity (40%), which correlated with the detection rate of emphysema. Free elastase activity and relative proteinase alpha 1-inhibitor deficiency suggest that the BALF protease-antiprotease system is impaired in patients with CDB and PC, which is more pronounced in patients with CDB. PMID- 14524101 TI - [Specific changes in the phagocytic activity of leukocytes in children after BCG vaccination]. AB - In 185 children vaccinated with BCG, the phagocytic activity of leukocytes (PAL) was studied in different postimmunization periods. The modified procedure for examining PAL that reveals and assays specific phagocytic changes irrespective of the level of PAL was used. A suspended matter mixture of BCG mycobacteria (a specific object) and white saprophytic staphylococci (a non-specific object) was used as the objects of phagocytosis. A specific increase in PAL in the first 1.5 years after vaccination and the specific changes of phagocytosis during a vaccination process were first established. There was a close relationship of the specific changes of phagocytosis to the size of an infiltrate at the site of vaccination injection. The strain of antituberculosis immunity may be more objectively assessed by the phagocytic test than by the size of a skin scar and by the Mantoux test with 2TE PPD-L. The phagocytic test is recommended for individual monitoring of a antituberculosis vaccination process in children. PMID- 14524102 TI - [Neurosensory deafness: immunological aspects]. AB - Seven hundred and twelve patients with first diagnosed pulmonary tuberculosis were examined; neurosensory hypoacusis was detected in 12.2% of the cases. The above patients underwent neurological, otolaryngological, and immunological studies. Second- and third-degree hypoacusis was found in 13.7 and 86.3%, respectively. Neurological syndromes were identified. These included vegetovascular dystonia in 50.5% of the patients, multiple cerebral minor symptoms in 45.5%, focal cerebral lesion in 4%/Comparison of the severity of nerve system disease and the level of hypoacusis has indicated that the more severe neurological disorders, neurosensory hypoacusis becomes more aggravated (K = 0.57). Positive skin Mantoux tests in all patients with neurosensory hypoacusis showed their allergization to Mycobacterium tuberculosis. In the examined patients, specific basophilic deformability (SBD) rates in the tuberculin test were 17.5 +/- 1.3% g, which differed significantly (p < 0.001) from those in the controls (5.1 +/- 0.9%). In second-degree neurosensory hypoacusis, SBD rates were significantly higher than those in first-degree one, which suggests that the allergic factor contributes to the development of neurosensory hypoacusis in the presence of tuberculosis. The values of antibodies to myeloperoxidase (1.869 +/- 0.02 optical density units (ODU) and to IgE (416.386 +/- 0.391 I.U./ml) significantly differed from those in the control group (0.08 +/- 0.02 ODU and 96.667 +/- 0.441 I.U./ml, respectively), which is indicative of primary lesions of minute blood vessels as the type of vasculitis and secondary nervous tissue involvement. Thus, allergic, vascular, and immune mechanisms are the main pathophysiological ones of neurosensory hypoacusis in the presence of first diagnosed and untreated tuberculosis. PMID- 14524103 TI - [Acute tuberculous bronchiolitis in a patient with generalized miliary tuberculosis]. AB - A case study of an acute tuberculous bronchiolitis (a rarely encountered form of generalized miliary tuberculosis), whose aggravated progressing resulted in fatal outcome 2 months after the onset of the initial disease signs, is described. The prevalence of non-specific infectious-and-allergic manifestations is a clinical and-morphological peculiarity within the case study, which confirms a diversity of morphological changes occurring in the organism in tuberculosis infection. The etiological- and pathogenetic specific features of acute tuberculous bronchiolitis in children and in adults are under discussion; in particular, a possible role of a preceding viral infection is evaluated. It is one of the cases of hematogenic generalization in primary tuberculosis of adults, observed by the authors, when an involvement of the pulmonary and parenchymatous organs as well as of arachnoids and substances of the cerebrum was registered. The presence of generalized vasculitis with fibrinoid swelling and with necrosis of collagenous fibers of medium (and with the development of pan-vasculitis in the discussed case) is typical of all such observations. PMID- 14524104 TI - [Epidemiology of extrapulmonary tuberculosis]. AB - The review paper presents data on the epidemiological features of extrapulmonary tuberculosis in Russia and other countries and on diagnostic problems. It considers the papers of the authors who propose different measures to enhance the efficiency of diagnosis of extrapulmonary tuberculosis. PMID- 14524105 TI - [Unknown pages of the history of organization of the Soviet phthisiological service]. PMID- 14524106 TI - [Proceedings of the Symposium on "Development of antituberculosis care for the population", March 17, 2002, within the framework of collegium of the Ministry of Health of the Russian Federation. Results of scientific studies "Phthisiopulmonology" for 2002]. PMID- 14524107 TI - [Pediatric tasks related to children's health protection]. AB - The activity of research facilities in shaping up the health of children and teenagers is elucidated. The priority trends are defined for joint research in the spheres of age-related and clinical physiology, psychology, and pediatrics as well as of hygiene of children and teenagers; as a result of such research, real methods can be suggested to monitor the health shape-up and development of children. PMID- 14524108 TI - [Health condition and medical care of adolescents in the Russian Federation]. AB - A dynamic description of teenagers' health status in the Russian Federation is presented in the article over a period of 40 years. A pronounced increasing proliferation of impaired functions and a prevalence of chronic pathologies in children were outstripping the growth rates; a worsening health condition in children revealed through a sharply reduced quantity of healthy children and a bigger share of chronically sick children; a changed structure of prevalence of functional disorders and of chronic pathologies among children reflecting, to a certain extent, the negative impact produced by the social as well as by ecological-and-hygienic conditions of contemporary life of schoolchildren; as well as a pronounced influence exerted by such significant factors, as social well-being, an intensified educational process and a premature labor occupation of children, on the evolution of children's health can be attributed to specific features of the build-up of children's health during different time periods under the conditions of today. The negative dynamics of the health status in children of the country necessitated a decision on reforming the provision of teenagers with medical care and on creating a system of continuous medical check-ups for them. Various forms of medical care as applicable to teenagers are analyzed. PMID- 14524109 TI - [Physical development of schoolchildren in the city of Moscow during the recent decade and a related assessment]. AB - Data on the long-term and short-term monitoring over the growth and development of schoolchildren in the city of Moscow during the second half of the last century are presented (including around 10,000 persons). According to the purpose oriented 40-year observations of the growing generation in Moscow, the accelerated physical development started in the 60-ies with its peak falling on the middle 70-ies; the elements of deceleration set on in the 80-ies and continued through the 90-ies. As for now, a deficit of the body weight, reduced width and span parameters and a decreased power abilities as well as shifts of the menarche time in girls towards an older age are typical of contemporary schoolchildren in Moscow. The detected trends are common regularities typical of children in all Russia's big cities. PMID- 14524110 TI - [Methodology of correcting the bearing in children and adolescents]. PMID- 14524111 TI - [Health status of adolescents and modern approaches to holding medical professional consultations]. AB - Due to unfavorable trends in the health state of contemporary children and teenagers, the total number of young males and females, who have health-related limitations in educational and occupational spheres, went up to 80%. A medical professional consultation belongs to a system of measures, whose purpose is to assist the young males and females with impaired health to choose professions or occupations, which would maintain their health intact and promote successfully their adaptation to professional training conditions and subsequently to actual occupational conditions. A timely and high-skilled implementation of medical professional consultations with a subsequent rational employment of teenagers and youth with impaired health should be regarded as a secondary prevention of pathologies. The holding of such medical professional consultations for teenagers is reasonable both at the stage, when a teenager makes his or her choice of a job (i.e. at school), and at the stage of receiving a professional education, so that the schoolchildren and students could get a job in line with their health status and that they could be employed with respect to doctor's recommendations. The Research Institute for Children's Health elaborated a number of methodological and regulating documents to promote the efficiency of the medical professional consultations; an automated assessment system is in the process of construction at the above institution. PMID- 14524112 TI - [Hygiene of shaping up a healthy working generation under the conditions of today]. AB - Prerequisites of shaping up the health condition, including health status, life mode and conditions of occupational training, in teenagers receiving workers' skills at basic-stage professional training facilities are under consideration. The specificity of adaptive responses and of health status is described in young females, mastering the jobs within the confectionary and public catering industry, during the professional training period, which is indicative of a higher sensitivity of teenagers' organisms towards the production factors' effects. Reliable trends towards aggravated indices of the health status in pupils of professional training facilities were registered during a 10-year observation period. A continued worsening of the life mode is noted in teenagers, which manifests itself mainly through a higher prevalence of smoking and frequent abuse of alcohol with the bad habits setting on at an earlier age. Labor occupation of pupils, when combined with professional training, is an extra risk factor in the building-up of health. Thus the obtained data witness to a worsening life mode of teenagers, who earn on the side, and to their worsening health status. Conclusions are made on a need to overcome the negative prerequisites related with shaping up the health condition of teenagers in the discussed social category. PMID- 14524113 TI - [Hygiene of contemporary preschool education]. AB - The case study deals with the health status of children at preschool educational facilities as observed in the 80-ies of the last century and early in the new century; besides, an elaboration of hygienic regulations applicable to the training-and-education effects produced on organisms of contemporary trainees of such facilities for as long as 3 to 7 years with due respect to the education modernization campaign (now underway in the country) is in the focus of attention. PMID- 14524114 TI - [Atopic disease in children: present-day concepts of pathogenesis and therapy]. AB - A contemporary understanding of evolution mechanisms and of therapy related with allergic diseases in children is defined on the basis of published data and independent research. A concept predetermining the significance of atopy in pathogenesis of allergic pathologies is outlined; an essential role of environmental factors and a genetic contribution to the role promotion are pointed out. Issues pertaining to the onset of both early and late phases of the allergic response and of allergic inflammation are touched upon. Finally, modern approaches towards therapy of allergic diseases in children are described. PMID- 14524115 TI - [Systemic and andragogic issues related to professional training of medical personnel in the sphere of hygiene of children and adolescents]. AB - The continuous education concept presupposes that a person is educated during his or her entire conscious lifetime. The pedagogical education model is made use of at the higher-school stage; while at the postgraduate stage, the andragogic model must be involved. The andragogics is a branch of science, according to which the leading role belongs to the person who is educated. He or she contributes extensively to the planning, organization, implementation, assessment and amendment of the educational process. Such process is translated into reality at the chair for hygiene of children and teenagers in accordance with the State Standard of postgraduate professional training of experts with higher medical education in the sphere of "hygiene of children and teenagers". The defined educational levels as well as psycho-physiological and specific features of the educating staff are a basis for a individual approach to them. Not only hygienists but also pediatricians and attending doctors from the chair for family life are trained at the above-mentioned chair. The diversity of training appicable to medical personnel of different categories necessitates serious educational-and-methodolgical measures. PMID- 14524116 TI - [New methodological approaches to the hygienic standards of clothes for children and adults]. AB - The hygienic assessment is presently a research issue related mainly with commissioning new materials into production; the above issue is equally related with materials' diversity, with expanding the choice of commodities and with the multi-functionality of the latter as well as with modern technologies and with the forthcoming admittance of our country to the WTO, which necessitates a harmonization of regulating requirements in various countries. The old approaches towards regulations of cloths are outdated and are no longer in line with the requirements of today. The chemical stability of contemporary synthetic fibers and the narrowing-of-gap between their physical-and-hygienic indices and those of natural fibers ensures unified approaches towards the hygienic evaluation of both. A real load that is conceptually found as a product of the impact intensity on the impact duration time is a hygienically valuable criterion ensuring a universal approach towards regulating the cloths hygienic safety. A hygienic classification provides, in case of each item, for defining the index of its hygienic safety (HIS), which is a basis for designing the differential requirements towards cloths of various functional purposes and for various age groups. The area of contact with the skin during wear and the age of consumer are the key components of system (The Hygienic Classification of Cloths". The article is supplemented by 1 drawing, 2 tables and a list of literature (8 entries; article's volume is 5 pages). PMID- 14524117 TI - [The intima -- a biological sorption filter. The specificity of pathogens and biological classification of inflammatory lesions in the intima]. PMID- 14524119 TI - [F, F. Talyzin and the Soviet parasitology (the 100th anniversary of his birth)]. PMID- 14524118 TI - [Neurobiological fundamentals of re-myelinogenesis in the central nervous system]. PMID- 14524120 TI - [Passive absorption of unsaturated fatty acids by cells. Fatty acids transporting proteins in cell cytosol]. PMID- 14524121 TI - [The role of hormone analysis in the nosologic diagnosis and in the control of treatment of congenital primary hypothyroidism]. AB - A total of 125 children and teenagers, aged 1 to 15 (including 74 patients with primary congenital hypothyroidism--CH--and 51 healthy children), were investigated in the city of Moscow for the purpose of elaborating new approaches towards the differentiated diagnostics of CH and of optimizing the assessment of the adequacy of its substitutive hormonal therapy (SHT). The concentrations of the thyrotropic hormone (TTH), free fractions of triiodothyronine and thyroxin (cT3 and cT4), thyreoglobulin (Tg) and of thyreoid peroxidase (hard-phase test tube method, electro-chemiluminescent reaction, i.e. magnetic microparticles, based on the streptovidin-biotin technology; ruthenium mark at the "Elecsys 1010" analyzer with reagents manufactured by "F. Hoffman--La Rosh Ltd.", Switzerland.) were determined early in the morning on the empty stomach. On the basis of the above mentioned, the thyroid reserve index (TRI = Tg/TG), integral thyroid index (ITI = [cT3 + cT4]/TTG), and the integral of peripheral conversion (IPC = cT3/cT4) were calculated. The thyroid gland (TG) was visualized by the "Acuson 128 XP" ultrasonic device, "Acuson Corp.", USA, with a linear sensor of 10 Mhz, as well as by the "Toshiba-90B" gamma-chamber, Japan, with a low-energy collimator in 24 hours after an oral intake of sodium iodide marked by the iodine 123-isotope with an activity of 50 Mbc and after the cessation of SHT 3 to 5 weeks before. The complex diagnosis revealed 8 variations of primary CH; aplasia and TG dystopia and defects of the synthesis of the sodium-iodine symporter or Tg concentration were found to belong to the most severe disease types according to hormonal parameters. As for Tg, in 52% of cases it was normal, undeterminable- 23% and higher--25%. A simultaneously implemented ultrasonic scanning (USS) ensured a final diagnosis in subgroups, respectively, in all cases, in 25% and in 75% of patients. On the whole, after the two diagnostic stages (Tg + USS/ITR), the indications for isotope scanning left only for 23% of CH patients, i.e. a) if the Tg values could not be determined because the organ was not located in its typical place (17%); or b) if the Tg level was higher or if there was a goiter at the TG normal volume (6%). An in-depth hormonal analysis showed that, should CHT be monitored less than once in three months, it failed to secure a total compensation of the disease in above one third of CH children (39%). Therefore, a three-stage algorithm of nosological diagnostics is recommended for primary CH; it should comprise the determination of Tg level, and USS (for all patients) as well as TG scanning (according to rare indications). The result point at the necessity of hormonal monitoring of CH children and teenagers at least one in 2-3 months. The objectivity degree of CHT goes up as a number of estimated indices, i.e. ITI and IPC, are used alongside with absolute hormonal values. PMID- 14524122 TI - [Age characteristics of free radical oxidation of lipids and antioxidant defence in erythrocytes from healthy individuals]. PMID- 14524123 TI - [Comparison of albumin analysis by fluorescent method with other clinical laboratory parameters in purulent peritonitis]. AB - The concentration and properties of the blood-albumin linkage centers were examined by the method fluorescent probes, besides, the general clinical-and laboratory indices in the blood serum of patients with diffusive purulent peritonitis were determined. The parameters as obtained by the method of fluorescent probes revealed a valuable information on a patients' severity condition and on a forecasted disease evolution; finally, they surpass, in a majority of cases, the generally accepted clinical-and-laboratory tests applicable to similar conditions. PMID- 14524124 TI - [Comparative characteristics of changes in the fatty acid spectrum of high density lipoproteins in male patients of various age with appendicitis]. AB - The fatty acid spectrum, the content of cholesterol of total high-density lipoproteins (HDLP) and high-density lipoproteins3 as well as the activity of lecithin-cholesteryl-acyl-transferase (LCAT) were investigated in mature-age males (periods I and II) with appendicitis. The intensity of endogenous production of polyunsaturated fatty acids was concluded to be reducing in mature age (periods I and II) males. As for the mature-age males of period I, the consumption of polyunsaturated fatty acids was found to be increasing and the production of endogenous cholesterol was found to be decreasing in them. The transformation of HDLP3 into HDLP2 was found to be declerating in the mature males of period II, while the cholesterol-acceptor ability of HDLP3 was going up in the males of latter category. PMID- 14524125 TI - [Cytological diagnosis of bone neoplasms in children (lecture)]. PMID- 14524126 TI - [The role of stromal and hematopoietic tissue interaction in pathology of hematopoiesis]. PMID- 14524127 TI - [Morphometric and biochemical analysis of erythrocytes in smoking patients with late vascular complications of diabetes mellitus]. AB - The following phenomena are observed in patients with late vascular complications of diabetes mellitus (DM): the smoking habit alters the quantitative and morpho functional characteristics of the erythron cells; it inhibits the lipid peroxidation processes in the erythrocytes membranes and aggravates the clinical course of endotoxicosis, which has a pathological impact on the rheological blood properties and worsens the tissues' hypoxia. A correlation was found between the pronouncement degree of anemic syndrome in diabetic angiopathies, on the one hand, and smoking, on the other hand. Smoking was found in patients with diabetic angiopathies to be a factor enhancing the metabolic stress, early signs of vascular complications and a risk of developing the anemic syndrome. PMID- 14524128 TI - [Express-diagnostic methods in urolithiasis for medical check-ups and screening]. AB - The suggested tool's set designed for express-diagnosis of urolithiasis and for ultrasonic screening of the kidneys, which comprises a new diagnostic technology ("Litos-System'), provides for stating a diagnosis even before the renal calculi begin to take shape. PMID- 14524129 TI - [Significance of the computer-assisted image analysis of nuclei as an additional tool for diagnosis of thin-needle aspiration biopsy of specimens from liver neoplasms]. PMID- 14524131 TI - [Etiology of purulent infections in the Tiumen' region based on hospital data]. AB - A study of laboratory findings for 474 examined patients, who were discharged from the Surgical Department of the Tyumen Regional Clinical Hospital, showed that S. aureus caused the infectious processes in 31.7% of the examinees. Oxacyllinresistant strain S aureus made up 9.7%.e examinees, and 22.6% of pathologies were caused in them by E. coli. E. coli was frequently found to be combined with polyresistant strains E. faecalis, E. faecium, K. pneumoniae and P. aeruginosa, which could be explained by the circulation of hospital strains within a hospital and by the selection of microorganisms with a high resistance level to antibacterial drugs. PMID- 14524132 TI - [Standards and errors in determining the sensitivity of bacteria to antimicrobial agents by disk-diffusion method]. PMID- 14524133 TI - Elderly suicides: a need for prevention. AB - Every 18 minutes a life is lost to suicide. Approximately every hour and a half, an elderly adult age 65 and over takes his/her life by suicide. Despite a decline in the rate of suicides over the past several years, suicides by elderly white males remains high. This paper provides a descriptive epidemiology of elderly suicides in a southern rural population and discusses implications for preventive health services that must be implemented to decrease the number of successful elderly suicides. PMID- 14524134 TI - After-session with George E. McGee, MD, 2003-04 MSMA president. PMID- 14524135 TI - Looking back: a forgotten use of calomel. PMID- 14524136 TI - Meth--it's everybody's problem. PMID- 14524137 TI - Prevnar--will it be the next vaccine success story? PMID- 14524138 TI - Chronic sorrow. PMID- 14524139 TI - Web-based classroom data collection in ADHD: a best practices case study. AB - BACKGROUND: Best practices research identifies and disseminates effective clinical strategies developed and refined by clinicians for their practices. As opposed to the content of health care, medical students and residents are typically not trained in the process of its provision. This paper illustrates a best practices approach to development of a Web-based classroom data collection method to improve medication management in attention deficit hyperactivity disorder (ADHD). OBJECTIVES: 1) To increase Primary Care Physician (PCP) awareness of the process of best practices research, and; 2) to highlight the use of the World Wide Web as a tool for integration of care of children with ADHD. METHODS: Based on the review of a large national study in ADHD, the key ingredients for the best pharmacotherapy practices were identified. The lessons from the literature were adapted for local practice using the framework and steps for best practices outlined by Mold and Gregory. A conceptual model of managing information provided by parents/caregivers and teachers to PCPs was developed. Several steps were identified, including Web-based classroom behavioral data collection by a care manager and feedback reports to PCPs from a child psychiatrist based on the data collected. This model is currently being tested. CONCLUSION: Our preliminary experience suggests that a best practices approach has potential applications to a variety of primary care settings in South Dakota. We encourage PCPs to consider best practice strategies for their own practices. The practice-based research network (Rush Net) being developed by the Center for Rural Health Improvement of the University of South Dakota School of Medicine's Department of Family Medicine can be helpful in this regard. PMID- 14524140 TI - [Apathy syndrome: a clinical entity?]. AB - Apathy is defined as a disorder of motivation that expresses itself at an emotional, cognitive and behavioural level. Apathy can occur as a symptom and a syndrome. In the recent years diagnostic criteria and a number of scales for measuring apathy in elderly with psychiatric or neurological disorders have been introduced. Two scales are specifically developed to measure apathy, the Apathy Evaluation Scale (AES) from Marin and the Apathy Scale (AS) from Starkstein. Both scales have been translated into Dutch. The AS is more convenient. The AS in addition can be used when applying the criteria for the apathy syndrome which has been introduced in 2001 by Starkstein. In addition, the Neuropsychiatric Inventory (NPI) and the 'Gedragsobservatieschaal voor de Intramurale Psychogeniatrie' (GIP) (a scale in Dutch) have an apathy domain. Conceptual problems surrounding apathy have only partly been resolved. The criteria for the apathy syndrome can only be used for assessing the extent of the problem. Apathy and depression are strongly correlated. Studies show that apathy as a syndrome can occur without concomitant depression in the elderly, but regularly occurs besides a depressive disorder, in percentages varying between 9% and 53% of the population under study. Especially the varying validity of an apathy syndrome in relation to late life depression needs further clarification. PMID- 14524141 TI - [Effects of emotion-oriented care on elderly people with cognitive impairment and behavioral problems in residential homes]. AB - This study investigates the effects of emotion-oriented care on the behavior of elderly people with cognitive impairment and behavioral problems. This approach is mainly based on the validation approach, but uses also insights from other approaches like reminiscence and sensory stimulation. Sixteen homes for the aged with structured day care units were randomly allocated to an intervention or control group. 151 Residents with cognitive impairment and behavioral problems were included in the study. The eight intervention homes received a training program with regard to emotion-oriented care. In the eight control homes usual care was continued. Measurements were performed at baseline and after three, six and twelve months of follow-up (assessment by caregivers and relatives). The primary outcome measure was the change in behavior of the residents. The results of multilevel analyses (overall, subgroup and per protocol) showed no statistically significant, nor clinically relevant effects in favor of the intervention group on the behavioral outcome measures. A possible reason for this is that the intervention did not result in significant contrast between the study groups. There is insufficient scientific evidence yet to justify the implementation of emotion-oriented care in residential homes, for residents with moderate to severe cognitive impairment and behavioral problems, on a large scale. Additional studies are needed in which special attention should be given to the implementation process. PMID- 14524142 TI - [The health status and health care use of elderly living in sheltered residences. A comparison with independently living elderly]. AB - A sheltered residence for older persons is a living arrangement in between independent living and a residential institution. There is little knowledge about the health of elderly living in these residences. We studied the physical, functional and psychosocial health of 401 persons living in a sheltered residence for elderly in Zwolle through a standardised interview. We compared the health of these elderly to the health of independently living elderly in the same region studied in the Longitudinal Aging Study Amsterdam and adjusted for sex, age and married state. We found an impaired functional health and more chronic illnesses among elderly living in a sheltered residence. In addition, more medication was used (OR = 2.4). The studied elderly felt more often depressed (OR = 1.9) or lonely (OR = 1.7). Their mental state was more frequently impaired (OR = 1.7), they visited health professionals more often and received more help for personal care (OR = 2.0) and housekeeping (OR = 1.6). We conclude that people living in sheltered residences form a distinct group of elderly, characterised by impaired physical, functional and psychosocial health. Planners of future care must be aware of this vulnerable group. PMID- 14524143 TI - [Recall of the prime minister for the assessment of mental status]. AB - A number of cognitive screening tests assess knowledge of the head of state or current prime minister. It is supposed that correct recall is a valid indicator of cognitive functioning. A consecutive sample of 259 visitors of a psychogeriatric day care center were rated for knowledge of the prime minister's name. Recall of his name was not dependent on level of education. The mean score on an independent screening test was significantly higher for those who knew the prime minister's name than for those who failed to answer the question. More men than women correctly answered the question, regardless of education level, living arrangements (living with someone versus being single and living alone), age and cognitive score. Using item response theory and Mokken scale analysis it was found that a short screening test containing the prime minister item constituted a scale of medium scalability (Loevinger's scalability coefficient H:0.50). Scores on the screening test correlated significantly with two independent measures of episodic and semantic memory. Deletion or addition of the prime minister item did not influence the degree of association between screening test and memory test scores. It is advisable to ask for the name of the prime minister in screening for cognitive impairment. Passing or failing the prime minister item correlates with intact or impaired retrieval of current information. PMID- 14524144 TI - Laboratory approach to acute and chronic hepatitis. PMID- 14524145 TI - Anatomy of an exposure: a hospital lab's recovery of Brucella melitensis. PMID- 14524146 TI - Talking with Joe Crowley. Interview by Dottie Dunham. PMID- 14524147 TI - Competition and cooperation: labs in a complex industry. PMID- 14524148 TI - To test or not to test? PMID- 14524149 TI - The Belgian surgical malpractice survey 2003. PMID- 14524150 TI - Notorious pitfalls in the care of multi-trauma patients. AB - In this review we elaborate on the more specific circumstances that are needed for adequate trauma care, such as the correct recognition and management of a tension pneumothorax, a tracheobronchial disruption, systemic air embolism and hypoventilation. Furthermore the trauma clinician must be aware of the different life threatening causes of haemorrhage and hypovolemia. Traumatic pericardial tamponade, myocardial contusion and a tension pneumothorax can all prove to be difficult diagnoses, but may all present with signs of hypotension with an increase in central venous pressure (CVP). In contrast, internal haemorrhage is most often accompanied by hypotension with a low CVP. Immediate evaluation and treatment of thoracic trauma, such as rupture of the aortal arch, is mandatory, as is the utilisation of the correct diagnostic strategy to evaluate the possibility of intra-abdominal and retroperitoneal injury. Both an unnecessary laparotomy and a delayed diagnosis must be avoided when dealing intra-abdominal injuries, such as kidney trauma. Furthermore, we stress the importance of the swift diagnosis and treatment of fractures of long bones and the pelvis, to prevent ongoing massive haemorrhage. Certain criteria should be met, in a hospital setting with sufficient day-to-day trauma experience, to be able to provide quality care for the multi-trauma patient. This will minimise the risk of errors and serious medical and judicial consequences. PMID- 14524151 TI - The acute atraumatic peroneal compartment syndrome, a rare and therefore sometimes unrecognised entity. AB - Two cases of Acute Peroneal Compartment Syndrome, without history of trauma are presented. One case had a history of prolonged horse-riding with the wearing of tight boots, the other case had a history of prolonged walking. An Acute Peroneal Compartment Syndrome should always be considered in the differential diagnosis of pain in the peroneal region of the lower limb. PMID- 14524152 TI - Pitfalls in penetrating trauma. AB - In Western Europe the most frequent cause of multiple injuries is blunt trauma. Only few of us have experience with penetrating trauma, without exception far less than in the USA or South-Africa. In Rotterdam, the Erasmus Medical Centre is a level I trauma centre, situated directly in the town centre. All penetrating traumas are directly presented to our emergency department by a well organized ambulance service supported by a mobile medical team if necessary. The delay with scoop and run principles is very short for these cases, resulting in severely injured reaching the hospital alive in increasing frequency. Although the basic principles of trauma care according to the guidelines of the Advanced Trauma Life Support (ATLS) (1-2) are the same for blunt and penetrating trauma with regard to priorities, diagnostics and primary therapy, there are some pitfalls in the strategy of management in penetrating trauma one should be aware of. Simple algorithms can be helpful, especially in case of limited experience (3). In case of life-saving procedures, the principles of Damage Control Surgery (DCS) must be followed (4-5). This approach is somewhat different from "traditional" surgical treatment. In the Ist phase prompt interventions by emergency thoracotomy and laparotomy are carried out, with only two goals to achieve: surgical control of haemorrhage and contamination. After temporary life-saving procedures, the 2nd phase is characterized by intensive care treatment, dealing with hypothermia, metabolic acidosis and clotting disturbances. Finally in the 3rd phase, within 6 24 hours, definitive surgical care takes place. In this overview, penetrating injuries of neck, thorax, abdomen and extremities will be outlined. Penetrating cranial injuries, as a neurosurgical emergency with poor prognosis, are not discussed. History and physical examination remain the corner stones of good medical praxis. In a work-up according to ATLS principles airway, breathing and circulation should be evaluated with great care. Neurovascular examination related to trauma of the spinal cord, peripheral nerves as well as vascular involvement should be carried out also in extremity injuries. Physical examination should be completed by localization of all stabwounds, in- and outshot openings as well as recto-vaginal examination and inspection of the oropharynx. PMID- 14524153 TI - Traumatic rupture of the thoracic aorta. AB - Traumatic rupture of the thoracic aorta is a life threatening situation, and may be secondary to several mechanisms; mainly penetrating or iatrogenic lesions and blunt trauma. Although penetrating mechanisms predominate, the number of patients with aortic disruption due to blunt trauma has continued to increase. This paper shows an overview focusing on the pathogenesis, diagnosis, timing and type of treatment regarding traumatic injuries of the thoracic aorta; it also reports the experience of one single center that deals with these lesions. The major difficulty in the evaluation data on blunt aortic injury is that retrospective reviews often group together patients with all types of aortic lesions, comparing outcomes for injuries in different locations, with diverse methods of repair and different surgeons and/or institutions. PMID- 14524154 TI - Treatment of hydrofluoric acid burns. AB - Hydrofluoric acid injuries have a potential for both systemic as well as severe local tissue destruction. In this article the different treatment modalities will be presented. Hydrofluoric acid is frequently found in the semiconductor industry, in rust removers and facade cleansers. The negligence or carelessness of workers and ignorance of the risks of hydrofluoric acid promote the incidence of these severe burns. To prevent these burns, adequate information for the workers is necessary. Splash goggles and neoprene gloves as well as laboratory coats should be worn at all times to prevent eye and skin contact. In cases of exposure, therapy should be accurate and immediate. PMID- 14524155 TI - Vein graft surveillance: is the yield worth the effort? AB - OBJECTIVES: The aim of this review is to provide an overview of evidence in support of the routine use of duplex surveillance during the follow-up of vein bypass graft patients. METHODS: The following review describes the results from a summation analysis, and data from two small-scale randomised controlled trials. RESULTS AND CONCLUSIONS: The above review puts into question the need for routine graft surveillance for all. It is agreed that the result of the Vein Graft Surveillance Trial will clarify the future needs for surveillance. PMID- 14524156 TI - Tibioperoneal angioplasty and bypass. AB - Revascularisation by angioplasty or bypass of narrowing and occlusion of the tibio-peroneal segment is done infrequently compared with the more proximal arteries, particularly the superficial femoral artery. The indication for intervention is critical limb ischaemia and the pattern of disease when it predominantly affects the calf arteries is most commonly seen in diabetics. Surgical bypass is the orthodox technique, with well-documented long-term results. Balloon dilation has recently been tried, particularly using the sub intimal technique. The advantage of minimal invasiveness has to be set against the inapplicability of the method if severe calcification is present, as is often the case in diabetics, the high restenosis rate, and the absence of controlled trials showing good evidence of long-term patency. PMID- 14524157 TI - The outcome of gallbladder perforation during laparoscopic cholecystectomy. AB - PURPOSE: Laparoscopic cholecystectomy (LC) has become the standard treatment for symptomatic cholelithiasis. The perforation of gallbladder, bile and stone spillage, trauma to bile duct are the most common complications of LC. This study was carried out to assess the outcome of gallbladder perforation (GBP) during LC. MATERIAL AND METHODS: Between 1991 and 2001, intraoperative GBP occurred in 48 patients during LC among 336 patients who were operated on for acute cholecystitis in 14, and for chronic cholecystitis in 322. RESULTS: Perforation was observed in 48 patients (14%). Perforation occurred during traction and grasping in 36 patients (75%), dissection of gallbladder in 8 patients (16%), and removal of the gallbladder in 4 patients (8%). Bile spillage occurred in 40 patients (83%) while bile and stone spillage was present in 8 patients (16%). During the follow-up period, a patient was diagnosed with a subhepatic abscess on the 15th postoperative day and percutaneous drainage was performed. CONCLUSIONS: During LC careful attention must be taken in order to prevent perforation, and the procedure must be performed by experienced surgeons with adequate instrumentations. If perforation occurs, extensive prompt retrieval of the bile and stone spillage must be done along with abundant irrigation. In case of inadequate aspiration and irrigation, the patients must be closely followed. PMID- 14524158 TI - The effects of dimethylsulfoxide in experimental obstructive jaundice. AB - MATERIAL AND METHODS: Thirty rats were divided into three groups, as sham, control and DMSO groups. Laparatomy was performed on each animal in the control and DMSO groups and common bile ducts were ligated. Common bile duct was observed but was not ligated for the rats in the sham group. Saline solution injection (1.5 mg/kg/intraperitoneally (i.p.)) was begun on the first day of surgical procedure and repeated once a day for the next 5 days. The same procedure was performed with DMSO (1.5 mg/kg/i.p.) instead of saline in the DMSO group. The rats were sacrificed on the postoperative seventh day, at which time venous blood and liver tissue specimens were taken. MAIN OUTCOME MEASUREMENTS: On the 7th postoperative day, the bilirubin, AST, ALT, ALP and GGT levels of the control and DMSO groups were significantly higher in comparison with the sham group (p < 0.01). On the 7th postoperative day, the erythrocyte superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels of the control and DMSO groups were significantly lower than those of the sham group (p < 0.01), but there was no statistical difference between the two groups (p > 0.05). Erythrocyte and liver malondialdehyde (MDA) levels in the control and DMSO groups were significantly higher compared with the sham group (p < 0.01). However, the MDA levels were significantly lower in the DMSO group compared to the control group (p < 0.01). CONCLUSION: It is stated that free oxygen radicals seem to play a role in the liver tissue injury, secondary to obstructive jaundice. In our experimental study, exogenic DMSO seems to have decreased lipid peroxidation and to have improved some of the parameters of liver tissue injury due to the obstructive jaundice in rats. PMID- 14524159 TI - Chest wall invasion in lung cancer patients. AB - This study was performed to assess the prognosis in patients with non-small cell lung cancer invading the chest wall. In this study, the data from 43 patients who were operated on between January 1990-January 1998, for non-small cell lung cancer with pathologically verified parietal pleural and chest wall invasion were retrospectively reviewed. The median and 3-year survival of the population was calculated to be 16.8 months and 34%. The pathologic stages were T3N0 in 31 (72.09%) patients, T3N1 in 5 (11.62%) and T3N2 in 7 (16.27%). The median survival of the T3N0M0 patients was 24 months but in the same T3 population with pathologically verified N1 and N2, the median survival was 7.4 months (p < 0.01). A complete resection was achieved in 37 (86.84%) patients. The median and 3-year survival of the patients with complete resection were 20.60 months and 41% respectively. In six patients, who had incomplete resection, median survival was noted to be 7.4 months. Patients who received adjuvant radiotherapy in the N2 positive group and the incomplete resection group, did not benefit (p > 0.05). The results of this study confirmed that the lung cancer patients with chest wall invasion had different survival curves. The survival of patients changed according to the completeness of the resection and lymphatic metastases of either N1 or N2. PMID- 14524161 TI - Maggot therapy: many hands make light work. AB - We report the case of a diabetic patient with severe ischaemic infected ulcer of the right foot, successfully treated with maggot therapy. PMID- 14524160 TI - Surgical treatment of hydatidothorax. Series of 79 cases. AB - OBJECTIVE: A retrospective review of 79 cases of ruptured intrapleural pulmonary hydatid cysts. We analyse and evaluate our experience in the surgical treatment of this complication. MATERIALS AND METHODS: In a ten-year period, from 1990 to 1999, 79 patients were operated on in our service for intrapleural rupture of a pulmonary hydatid cyst. There were 51 males and 28 females with a mean age of 35.4 years. The diagnosis was established on the basis of different clinical signs and imaging studies. Surgical approach consisted of a posterolateral thoracotomy in all cases. After decortication, different procedures were performed on the pulmonary lesions according to the importance of lung destruction. RESULTS: Radical resections were done in 48 cases, including lobectomies (15), segmentectomies (33) and conservative treatment: simple capitonnage and bronchial fistula closure (31). Postoperative complications occurred in 8 cases (10.1%), including one pyothorax, one haemothorax, one prolonged air leak, two pneumonias and two wound infections. There was one postoperative death, by respiratory failure. Ninety-five percent (95%) of patients were free of recurrence of thoracic hydatid disease in a follow-up ranging from 1 to 10 years (mean: 5.4 years). CONCLUSION: Hydatid cysts of the lung should be treated before complications occur, particularly intrapleural rupture because it considerably increases morbidity. PMID- 14524162 TI - Sclerosing peritonitis: report of three cases. AB - Sclerosing peritonitis is a dramatic complication of continuous ambulatory peritoneal dialysis and chronic peritoneal inflammation. Both visceral and parietal surfaces of the peritoneal cavity are involved. A thickened peritoneum encloses the small intestine in a "cocoon" formation which often leads to intestinal occlusion. CT scan may help obtaining an early diagnosis but diagnosis is often established with some delay or even at the time of laparotomy. Our report describes three cases of this uncommon peritoneal fibrosis syndrome which caused intestinal obstruction requiring surgical intervention. PMID- 14524164 TI - Primary peripheral adenoid cystic carcinoma of the lung. A case report. AB - We report a rare case of peripheral adenoid cystic carcinoma of the lung, showing unusual pathological and clinical features, namely rapid growth, local aggressive behaviour, huge tumour size, no endobronchial component or submucosal infiltration, and a rapidly progressive clinical course. Extensive surgery resulted in considerable palliation of symptoms, but not in prolonged survival. The reported case emphasizes the malignant potential of peripheral adenoid cystic carcinoma of the lung. PMID- 14524163 TI - Hepatocellular carcinoma associated with precocious puberty and oral contraceptives. A case report. AB - A 36-year-old woman presented with sudden abdominal pain and vomiting. Computed tomography showed a tumour of the right hepatic lobe with possible signs of acute haemorrhage. Her medical history revealed precocious puberty when she was a 5 year-old and the use of oral contraceptives for 18 years. Bisegmentectomy was performed and histological examination revealed hepatocellular carcinoma. The role of male and female sex hormones in the development of hepatic tumours has been well documented but, to our knowledge, association with precocious puberty has not yet been described. PMID- 14524165 TI - Gastroduodenal artery pseudo-aneurysm after cholecystectomy. AB - We report the case of a 69-year-old man presenting with rupture of a pseudo aneurysm of the gastroduodenal artery into the duodenum eight days after open cholecystectomy for acute cholecystitis. The surgical approach to a ruptured visceral artery pseudo-aneurysm is technically difficult and hazardous. The operative mortality rate reaches 16 to 50%, mainly dependent on the anatomic location of the visceral artery pseudo-aneurysm. This report describes a case in which bleeding from a pseudo-aneurysm of the gastroduodenal artery was successfully treated by embolisation using metal coils. PMID- 14524166 TI - Primary malignant melanoma of the oesophagus. AB - Primary malignant melanoma arising in the oesophagus is a rare condition with a dismal prognosis. The diagnosis is often made following surgical resection even though the endoscopic features may be pathognomonic. The classical treatment is oesophagectomy even though the advanced disease stage at the time of presentation and aggressive biological behaviour of the tumour usually results in a fatal outcome. We report the case of a male patient initially diagnosed with squamous oesophageal carcinoma and treated with conventional neo-adjuvant chemo radiotherapy. Poor clinical and radiological response resulted in a review of the original histology confirming a diagnosis of primary malignant melanoma of the oesophagus. The subsequent alteration in management conferred the patient an improved quality of life. A short review of the literature on primary malignant melanoma of the oesophagus supplements this case report. PMID- 14524167 TI - Perforation of the stomach due to chest tube complication in a patient with iatrogenic diaphragmatic rupture. AB - Diagnosis of diaphragmatic injury is difficult. A case of iatrogenic diaphragmatic rupture is reported in which perforation of a herniated stomach occurred following left lobectomy and partial resection of the diaphragm for lung cancer. On the second postoperative day, bile-stained fluid coming out from the chest tube revealed gastrointestinal leakage. This rare complication of chest tube insertion, early diagnosis and treatment are emphasized. PMID- 14524168 TI - Hypoplasia of the right hepatic lobe combined with a floating gallbladder. AB - Agenesis or hypoplasia of the right hepatic lobe combined with a floating gallbladder is an extremely rare condition. We report a case of hypoplasia of the right hepatic lobe, discovered in a 65-year old female. This was an incidental finding at CT scan for staging of a right colonic cancer. The CT evidenced the presence of a hypoplastic right lobe, while the left lobe was diffusely enlarged. Furthermore, the gallbladder was described as floating with partially calcified walls. The diagnosis of this rare anomaly was confirmed intraoperatively. The patient underwent right hemicolectomy and cholecystectomy. Biopsies were taken from both right and left hepatic lobes, revealing the presence of normal hepatic parenchyma. Since all causes of acquired atrophy of the liver had been ruled out, we considered this case to be of congenital origin. PMID- 14524169 TI - How can we approach a left-sided stab wound in the neck, with isolated tracheal laceration? AB - Any penetrating trauma to the mediastinum may cause great vessel damage resulting in massive bleeding or even shock. Associated tracheal injury increases morbidity and mortality. We report a case of a penetrating mediastinal injury with isolated tracheal laceration. The stab entered in the left supraclavicular fossa. The patient presented with right-sided pneumothorax, pneumomediastinum and respiratory collapse. Bronchoscopy revealed two injuries in the trachea that were repaired by right thoracotomy and left cervicotomy. The patient was discharged on postoperative day 6. Isolated thoracic tracheal injury is very rare in mediastinal stab wounds and operative strategy may change according to bronchoscopic findings. PMID- 14524170 TI - Leveraging hospital investment in ER services. PMID- 14524171 TI - Physician conflicts are tough to resolve. PMID- 14524172 TI - Texas health resources shows a proactive merger can pay off. PMID- 14524173 TI - Experts offer tips for merger success. PMID- 14524174 TI - [Survival of patients with laryngeal cancer and some prognostic factors]. AB - Tumour growth and its progression to a metastatic phenotype involves a serious of genetic events with abnormal activation of oncogenes or inactivation of tumour suppressor genes and others genes connected with proliferation, apoptosis and neovascularisation. The aims of the study were to determine the possible prognostic value of angiogenesis, proliferation index Ki67, p53 and bcl-2 proteins expression in patients with laryngeal cancer. The group of 151 patients with laryngeal cancer, surgically treated with minimum 5 years observation, was multi-variously analysed. Paraffin--embedded tissue sections from each case were stained with a monoclonal antibody raised against FVIII antigen, p53 and bcl-2 proteins and Ki67 proliferation antigen using a peroxidase labelled streptavidin- biotin kit in standard immunohistochemistry techniques. In univariate analysis: staging IV, tumour size T4, nodal metastasis N2 and N3, local and nodal recurrences, high expression of Ki67 and P53, high (over median) IA measured as number of microvessels with FVIII expression were significantly associated with shortened overall survival. Disease-free survival was related to: proliferation index Ki67, expression of P53 protein and angiogenesis measured as microvessels density with expression of FVIII antigen. In multivariate analysis the most important death risk factors for overall survival were: tumour size, nodal metastasis, local and nodal recurrences, P53 protein expression and IA measured as number of microvessels with FVIII expression. In multivariate analysis of disease-free survival only P53 protein expression, proliferative index Ki67 and expression of FVIII had independent prognostic value. Intensity of angiogenesis, proliferation index of Ki67 antigen and expression of P53 protein were independent predictors of patients with laryngeal cancer outcome. In contrary Bcl2 protein seems to be useless in these patients. PMID- 14524175 TI - [Laryngeal and hypopharyngeal carcinoma--the patient's perspective]. AB - Carcinoma of the larynx and hypopharynx are the most often head and neck malignancies. The make diagnosis of the disease early and established the best methods of treatment for individual patients is important just the same as a good relation between doctor and patient. The subject of the study has been patient's judgements about quality of diagnosis, treatment and follow-up procedures. 60 patients (10 women and 50 men) has been inquired with contents the same list of questions. All patients have been treated because of carcinoma of the larynx and laryngopharynx during last 5 years in the Department of Otolaryngology Medical Academy in Warsaw. The authors analysed all answers for questions. The general conclusions have been definite: the patient's knowledge about laryngeal and pharyngeal anatomy and functions is minimal, also they know not much about neoplasm disease and cigarettes and alcohol harmfulness. The patient's opinion at doctor's professional authority was good. It seems to be necessary to do much more to improve people health education especially wholesome behaviour. PMID- 14524176 TI - [Results of surgical and combined (surgery and radiotherapy) treatment in carcinoma of the larynx and hypopharynx--20 years experience of the ENT Department, District Hospital in Kielce]. AB - The epidemiological characteristic of 940 patients with carcinoma of larynx and hypopharynx, treated from 1978 to 1997 was presented, as well as the results obtained in this group with surgical and combined (surgery + rtg-therapy) treatment. There was a prevalence--75.2% of highly advanced cases (III degrees + IV degrees). The 65.6% of patients had been treated by surgery alone, and the remaining had received additional rtg-therapy after surgery. The 3-years survival rate had been achieved in 77.8%, and 5-years survival in 61.7% of the whole group. The analysis of the survival rates had proved the significant differences depending mainly to localisation of primary tumour, and clinical advancement stage of the disease. PMID- 14524177 TI - [Adjuvant radiotherapy of the neck lymph modes in patients after partial laryngectomies]. AB - Adjuvant radiotherapy after partial laryngectomy is still controversial mainly due to high level morbidities which may occur in the irradiated larynx. The aim of this study was to evaluate the results (locoregional control) and toxicities (acute and late) of combined treatment. Between 1996 and 2001 the group of 14 patients (12 men, 2 women), aged less than 60 years, Karnofsky scale > 70, were investigated retrospectively. The following surgical procedures were carried out: cricohyoidopexy (CHP) and cricohyoidoepiglottopexy (CHEP) in 9 out of 14 patients, horizontal laryngectomy in 1 patient, fronto-lateral laryngectomy in 4 patients. In all cases diagnosis of squamous cell carcinoma was confirmed. According to pathological classification the stage of disease in the neck was as follows: N1 = 8 patients and N2 = 6 patients. Indications for adjuvant radiotherapy were a high risk of microscopic spread of disease and expectation of regular follow-up problems in treated patients. The main rule of radiotherapy technique was avoiding the irradiation of the preserved larynx. All patients were irradiated postoperatively in conventional way to the elective dose of 50 Gy to the neck with additional dose of 16 Gy to the area of high risk of local recurrence. During follow-up no patients had locoregional failure. The toxicities (early and late) concerned the skin and mucosa of pharynx. These toxicities, evaluated according to the RTOG scale, in all cases were less than III grade. We conclude that combination of partial laryngectomy and postoperative radiotherapy with protection of the larynx during irradiation in the group of patients with high risk of regional recurrence plays an important role in decrease of treatment failures and permits preservation of laryngeal function without any significant side effects. Therefore the described method is an effective treatment for selected patients with the carcinoma of the larynx. PMID- 14524178 TI - [Nasal mucosa in patients with diabetes mellitus]. AB - Diabetes mellitus is the most common endocrinologic disease all over the world. 150 million people suffer from this disease, in Poland about 2 million. The disease on the basis of the onset and pathophysiology may be divided into type I and type II. Pathophysiologic changes include diabetic microangiopathy, macroangiopathy and neuropathy. The most common presentations in head and neck are otitis externa, hypoacusis, vertigo, disequilibrium, xerostomia, dysphagia, fungal and recurrent infections. The changes in nasal mucosa are not very well known. Only few papers concerned the problem. The main complaints of patients regarding the nose are xeromycteria, hyposmia and various degree of decreased patency of the nose. Chronic atrophic rhinitis, septal perforation, ulceration of nasal mucosa, alar necrosis, symptoms of staphylococcal or fungal infection can be found during otolaryngologic examination. The treatment in this group of patients should consist of systemic therapy of diabetes mellitus and on the other hand focal therapy with the use of a solution to moisten the nasal mucosa. PMID- 14524179 TI - [Use of allogenic cartilage in reconstruction of the middle ear in patients with chronic otitis media]. AB - We have analysed results treatment of 253 patients with chronic otitis media. Our tactic includes operation with application one-momental sanation and reconstruction of a middle ear. Sanational stage was performed in volume atticoantrotomia or radical operation. Reconstruction included reconstruction of a tympanic cavity and retrotympanic space. The shape of a plate was made so that to make a base for neotympanic membrane and to forma a lateral attic wall. Defect of a drum-type membrane restored free flap of a fascia temporal muscle. The mastoid process cavity was filled allogenic cartilage transplants the first variant of reconstruction formed artificial antrum cartilage by fragments and plates. At the second variant was acted similarly, however the artificial cavity wall of the antrum we made soft and mobile from the skin of external acoustical passage and neotympanic fascia flap. In postoperative period the position of a posterior wall of external acoustical passage was formed depending on intratympanic pressure, which depended on a acoustical tube function. Such technique allowed us to reduce formation retraction complications from 15% up to 6.7%. PMID- 14524180 TI - [Influence of tonsillectomy in adults on the concentration and glycosylation profile of acute phase proteins]. AB - Chronic tonsillitis (ch.t.) in adults can have the effect on immunological system and its function. The aims of this study were: 1. the estimation of concentration and glycosylation of acute-phase serum protein (a.ph.s.p.)--C-reactive protein (CRP), alpha 1-acid glycoprotein (AGP), alpha 1-antichymotrypsin (ACT), transferrin (Tf) and the concentration of immunoglobulins (Ig) and alpha 2 macroglobulin (alpha 2-M) in adults with ch.t. 2. the influence of tonsillectomy on the concentration and glycosylation a.ph.s.p. in I week and 6-8 months after surgery and determination of normalization estimated parameters. 116 patients with ch.t. and 15 healthy persons were evaluated. In all patients with ch.t. the concentration of acute-phase serum protein, alpha 2-M and immunoglobulins were determined before, in I-II day, VII day and 6-8 months after surgery. In patients with ch.t. were observed the significantly higher the concentration of CRP, AGP, ACT, IgG and the significantly decreased the concentration of serum transferrin before tonsillectomy, as an effects of immunological response on chronic inflammation. Tonsillectomy induced further increases the concentration of a.ph.s.p. and the highest value attained the CRP level. The change of glycosylation of AGP and ACT were results of chronic inflammatory process and analysis the glycosylation of these proteins can be useful in the estimation of activity of inflammatory process. The normalization of the concentration of acute phase serum protein were observed in 6-8 months after tonsillectomy and confirmed the withdrawal of inflammatory process. PMID- 14524181 TI - [Meningoencephalocele as rhinosurgical problem]. AB - Cephalocele is a rare lesion mainly of congenital or traumatic origin. The lesion occurs as an extracranial hernia of dura mater that consists of cerebrospinal fluid only (meningocele) or cerebellar tissue (encephalocele). Some cephaloceles concern anterior cranial fossa and then are observed as hernias in nasal cavity or in paranasal sinuses. Three patients are presented. In 52 year woman, who was admitted due to idiopathic rhinorrhea (persisting 16 years) with periodical headache, the meningocele in right ethmoid sinus was diagnosed. The lesion was removed with the rhinosurgical approach and the defect in anterior cranial fossa was repaired with the free cartilaginous flap. 12 year boy was admitted due to the nasal tumor diagnosed by CT and MRI. The lesion was removed with the combined access: firstly the hernia sac was cut intracranially with the frontal craniotomy and then encephalocele was removed with lateral rhinotomy. Cranial fossa defect was repaired by the periosteum flap. In third case (8 year boy) encephalocele caused 5 incidences of purulent meningitis. CT and MRI showed the lesion in frontal sinus. Encephalocele was removed with rhinosurgical approach and the defect in posterior frontal wall was repaired with free cartilaginous and mucosal flaps. In all patients long term result of operations was good. Anatomic and pathologic conditions that influence on the choice of rhinosurgical or combined access to nasal and sinusal encephaloceles are presented and discussed. PMID- 14524182 TI - [Evaluation of an animal model in endoscopic surgery of the cerebello-pontine angle]. AB - In the last few years one can observe a dynamic evolution of the minimal invasive surgery, particularly in the field of the skull base surgery. We are now in a front of a big problem with human specimen for dissection. They become rare. Those circumstances force us to look for an animal model to perform dissections. The swine is a very well known model for general and thoracic surgery but we can hear also some attempt of using it in the field of the skull base surgery. Authors tried to deal with the endoscopic anatomy of the cerebello-pontine angle through the retrosigmoid approach. During this study they found that we could find structures which are analogical to human's one. Animal models will not take place of human specimens in surgical training but they can considerably reduce their number. Difficult technique and unusual tools demands a lot of hours of training, and a big part of it can be easily performed on animal models. Authors present their own experience. PMID- 14524183 TI - [Anatomy of sinus tympani]. AB - 30 temporal bones from adult cadavers of both sexes were examined. After the tympanic sinus was identified, its morphologic features were evaluated. Minimum distance from adjacent structures: lateral and posterior semicircular canal, facial nerve canal and jugular fossa were taken. Measurements were taken under operative microscope with eye-piece graduation of 0.05 mm accuracy. Four main morphologic types of entrance to the sinus and two main developmental forms--deep and shallow sinus were distinguished. Deep sinus coexisted with absence of bridge and conversely if bridge was seen sinus was shallow. Deep sinuses were located close to the facial canal, in some cases penetrating around the canal up to 2/3 of canal's circumference. This anatomical variation increase the risk of the facial nerve damage during the surgery. In most of cases tympanic sinus is oval in shape. It's long diameter lies in vertical plane. The mean distance between the sinus and adjacent structures were as following: facial nerve canal--1.5 mm, lateral semicircular canal--2.1 mm, posterior semicircular canal--1.59 mm, jugular fossa--5.5 mm. No differences dependent from gender and side were found. PMID- 14524184 TI - [Spectral and spectrographic voice acoustic analysis in selected patients after various types of partial laryngectomy for malignant neoplasms after radiotherapy]. AB - The aim of this study is the assessment of voice pathology in patients after various types of partial laryngectomy because of malignant neoplasms and after Co -60--therapy. Voice quality in above mentioned diseases and in healthy patients was compared. Voice was analysed by subjective (videostroboscopy) and objective spectrographic methods. This method seems to be very valuable to assess social efficiency of voice. PMID- 14524185 TI - [Glasgow Benefit Plot for evaluation of stapedectomy result]. AB - In a group of 153 patients, the result of unilateral stapedectomy was evaluated using Glasgow Benefit Plot. Pure tone average at frequencies 0.5; 1; 2; and 3 kHz of operated and non-operated ear were used to distribute patients to pre- and post-operative groups. In 26 (79%) of 33 patients with unilateral hearing loss, bilateral normal hearing was achieved. Thirty one (46%) of 68 patients with asymmetric bilateral hearing loss and 37 (71%) of 52 patients with symmetric bilateral hearing loss had unilateral normal hearing after the operation. Twenty (29%) patients of the group III had bilateral symmetrical hearing loss after surgery. Stapedectomy was less beneficial for 17 (25%) of 68 patients with asymmetric hearing bilateral loss and 15 (29%) 52 of patients with symmetric bilateral hearing loss, who still had asymmetric hearing loss after the operation. Evaluation of hearing tests using the Glasgow Benefit Plot enables to evaluate patient's hearing disability and to predict possible benefit from surgery for individual cases. PMID- 14524186 TI - [Influence of drugs administered in general anesthesia and in pre- and post operative periods on transiently evoked otoacoustic emissions (TEOAE) in children operated for congenital heart defects (part II)]. AB - Outer hair cells are believed to be responsible for generation of otoacoustic emissions (OAEs). High sensitivity of OAE makes it possible to diagnose subclinical cochlear pathologies even if they have not yet been revealed by tonal audiometry. The aim of this study was to evaluate an influence of general anesthesia and other drugs administered in pre- and post-operative period on TEOAE in subjects operated on for various congenital heart defects in extracorporeal circulation. The investigated group consisted of 44 children with no pathology confirmed during ENT examination, in age ranging from 6 months to 19 years (average 9.3 years; +/- 5.9 years). The control group consisted of 14 otologically and cardiologically healthy subjects aged from 2-16 years (average 10.3 +/- 3.4 years). In the investigated group, TEOAEs were measured twice: one day before and 7-8 days following surgery. In the control group, TEOAEs were recorded in the same manner. Statistical analysis revealed no significant differences in the control group, between second and first TEOAE tests. Analysis of the influence of the drugs administered in general anesthesia did not show significant influence on post-operative TEOAEs. Among ototoxic drugs only acetylsalicylic acid affected TEOAE in post-operative tests. The authors conclude that general anaesthesia does not change the cochlear function, although, in children in whom acetylsalicylic acid was administered in post-operative period the cochlear function was affected. In these subjects the function of the cochlea should be monitored in post-surgical period. PMID- 14524187 TI - [Assessment of rehabilitation of teachers' voice disorders]. AB - Results of treatment of teacher's voice disorders were presented. Patients except pharmacological treatment and physiotherapy realised the course of voice emission exercises under speech therapist supervision. Ninety teachers were examined before and after treatment. Laryngological, phoniatric and laryngostroboscopic studies were carried out. In most patients phoniatric assessment revealed elongation of phonation time, changing of breathing route from cleidocostal to phrenocostal and voice creation without neck muscle tension after completing of treatment. 6 weeks lasting period of exercises was determined to be sufficient in most patients to learn correct breathing and phonation but to establish normal voice emission patient strong motivation, discipline and regularity during several months lasting exercises is needed. PMID- 14524188 TI - [Ectopic thyroid glad--a case report]. AB - Authors have described the case of 67 years women with an anterior midline neck tumor. The tumor was surgically excited and the microscope examination revealed thyroid tissue with colloid goiter. One must consider a midline ectopic thyroid gland in the differential diagnosis of masses in the neck. PMID- 14524189 TI - [Unusual case of a para- and retropharyngeal space tumor (cavernous haemangioma) obturating the laryngeal entry]. AB - The authors presents un unusual case of the para- and retropharyngeal space cavernous haemangioma displacing posterolateral wall of the pharynx to the laryngeal aditus. In the beginning tumor didn't cause any symptoms, but later life threatening dyspnea appeared. The surgical treatment with good anatomical and functional result is presented. PMID- 14524190 TI - [Angiosarcoma of the temporal bone]. AB - A case of 55-yrs old female was reported, in whom angiosarcoma of temporal bone had been diagnosed. A mode of treatment was surgery including partial petrosectomy and labyrinthectomy. Radiotherapy was not given because of a possibility of further cerebrospinal liquid leakage after dura mater rupture, which was seen during first operation. Local recurrence had developed about 2 months after surgery and a patient died 10 months after the time of diagnosis. It was stressed that angiosarcoma is a very rare neoplasm and in our case it was presented as few--symptomatic lesion despite of it's high extent in temporal bone. A review of literature concerning angiosarcoma was also reported. PMID- 14524191 TI - [Merkel cell carcinoma of the nose]. AB - Despite an increasing number of reports Merkel cell carcinoma still is a rare neoplasm. We therefore present the imaging feature of the case of Merkel cell carcinoma involving the nasal region in 65-year woman with lymphoma malignum lymphoplasmocytoides and its rapid progression with fast metastasis to lymph nodes. Surgery and adjuvant radiotherapy, chemotherapy appeared to provide optimal local control. PMID- 14524192 TI - [Remarks concerning up to date TNM staging of cancer localized in lip and oral cavity, pharynx, nasal cavity and paranasal sinuses, and major salivary glands]. AB - The general principles and historical background of TNM classification was briefly discussed, with advantages as well as weakness and limitations of the system. The current TNM staging of lip and oral cavity, pharynx. The currant TNM staging of carcinoma in lip and oral cavity, pharynx, larynx, nasal cavity and paranasal sinuses and major salivary glands was presented, pointing out the last changes in the classification. PMID- 14524193 TI - [Remarks on the hearing theory]. AB - Many remarks have been presented on the present-day theory of hearing. Some mechanisms connected with transforming auditory information have been discussed. One's own opinions have been presented on the possibility of existence of proteins which change their conformation when influenced by the energy of the sound wave. The mechanism mentioned above resembles the one existing in the organs of sight, smell and taste. PMID- 14524194 TI - [Conservative therapy of mastoiditis and therapeutical indications of mastoidectomy until the end of XIX century. The achievements of Varsovian physicians]. AB - The problem of treatment of the mastoiditis was discussed long ago--in XVII and XVIII centuries by Jean Riolan (1580-1657), Werner Rolfinck (1599-1673), Jean L. Petit (1674-1750), Jasser and the others. The therapeutic indications of mastoidectomy in 1873 by Hermann Schwartze (1837-1910) is mentioned. The achievements of such German physicians as Friedrich Bezold (1842-1908) and Ernst Kuster (1839-1930) are described in detail. The achievements of such Varsovian physicians as Teodor Heiman (1848-1917), Andrzej Alfred Heiman (1984-1933), Edmund Modrzejewski (1849-1893), Samuel Meyerson (1851-1939) and Ludwik Guranowski (1853-1926) are also presented. PMID- 14524195 TI - Statement on the recommended use of monoclonal anti-RSV antibody (palivizumab). PMID- 14524196 TI - National policy conference on antibiotic resistance, 5-7 October, 2002: summary of proceedings. PMID- 14524197 TI - Antimicrobial resistance: a deadly burden no country can afford to ignore. PMID- 14524198 TI - Terrorism and violence. Toward a preventive approach. PMID- 14524199 TI - Treatment of an earlobe keloid. PMID- 14524200 TI - Working in a war zone. The impact on humanitarian health workers. AB - BACKGROUND: The work challenges faced by doctors, nurses and other health professionals in the humanitarian field are overwhelming. OBJECTIVE: This article highlights the psychological effects on humanitarian workers and the support available, both while on a 'mission' and on return home. DISCUSSION: It is impossible not to be psychologically affected by witnessing gross acts of violence, starvation, epidemics, displacement and despair, or hearing tales of slaughter, rape and killing. Just as those populations who are subjected to traumatic experiences develop post-traumatic psychological problems, so too can those humanitarian workers who assist them. PMID- 14524201 TI - Post-traumatic stress disorder. A brief overview. AB - BACKGROUND: Post-traumatic stress disorder (PTSD) is an anxiety disorder which occurs following exposure to a traumatic, potentially life threatening event. It is frequently comorbid with other anxiety and mood disorders, substance misuse and physical symptoms that may cloud its presentation. In itself, PTSD is a frequently chronic disabling condition associated with a marked impact on social, occupational and family functioning that also carries an increased risk of suicide. Early intervention once the disorder is present, represents the most effective chance of reducing disability. Appropriate diagnosis and referral are among the key skills required by the general practitioner to deal with this condition. OBJECTIVE: This article aims to provide an overview of PTSD for GPs including clinical features, epidemiological aspects, approach to assessment and treatment, and specialist resources available. DISCUSSION: Recent world events including terrorist attacks, wars in Afghanistan and Iraq, and Middle East turmoil have raised community awareness of the impact of trauma. General practitioners are in a unique position to be involved in the early diagnosis and treatment of PTSD, which has been shown to improve outcome. PMID- 14524202 TI - Bali 12 months on.... Providing for continuing care and recovery. AB - BACKGROUND: Traumatic events such as the bombings in Bali may cause many people to develop post-traumatic stress, traumatic grief, anxiety or depression. OBJECTIVE: This article outlines the requirements of recovery, indications of its progress and the role of general practitioners in preventive care, support and referral. DISCUSSION: Research indicates that more than half of those involved in traumatic events can expect to recover without developing a disorder. However, even optimal recovery from tragedy is a protracted and arduous task that severely taxes physical, emotional and social health. The consequences of this may show as degraded health in the second year following a tragedy. PMID- 14524203 TI - Treatment of common 'life event' traumas. AB - BACKGROUND: General practitioners are confronted daily with patients who have experienced distressing 'life events'. Assisting patients through this process can present a significant challenge. OBJECTIVE: This article points to the reactive symptomatology that may be overtly present or concealed in a stressful event and its aftermath reactions, sets the goal of restoring a person's functioning and their self resources, and gives guidelines of what counselling can do for a patient. DISCUSSION: People who are attempting to cope with a distressing 'life event' or the aftermath of a trauma can be effectively helped by early short term counselling. Although this task may not be one that appeals to the busy GP, many accessible interventions can restore a person's coping capacity and avoid the prospect of a continuing and deepening pathology. Many symptoms will not fit the diagnostic categories usually associated with stress disorders. Some may show impaired social engagement and social phobia; others may describe low self worth, states of unwellness, relationship dissatisfaction and maladjustments in key areas of behaviour. The task is to help the person gain a different perspective on the distressing event, their own capacities and the supportive influences around them. PMID- 14524204 TI - Grief and loss. 'I'm not myself anymore'. AB - BACKGROUND: Most individuals who experience the loss or potential loss of a person or thing to which they ascribe importance in their lives undergo the process of 'normal grief'. Simply put, the experience of grief is not limited to the loss of a person through death, but is also felt when other life changes and transitions occur. OBJECTIVE: This article aims to address what is 'normal' grief while examining some strategies that can be useful in assisting the grieving indivudual. DISCUSSION: Current research asserts that there is no actual conclusion to the grief process, but rather an integration of the grief into an individual's life. To achieve a healthy integration of loss and attain the ultimate wellbeing of the individual, the grieving person may need to be assisted and guided in a biopsychosocial framework. PMID- 14524205 TI - Patient education. Understanding negative emotions. PMID- 14524206 TI - Writing the prescription and informing the patient. AB - BACKGROUND: Having decided on the generic drug to prescribe the prescription needs to be written and the patient informed regarding the treatment. OBJECTIVE: This article discusses some regulatory requirements for writing a legal prescription, but predominantly discusses important aspects of informing patients regarding drug therapy. DISCUSSION: The actual writing of a prescription is a small part of the prescribing process and needs to comply with local regulations. Including patients in the decision to embark on a treatment and informing them of drug treatment is a very important part of the prescribing process. There is no simple formula, but patients need to be informed on what they want to know, when they want to know it, and in the way that is most acceptable to them. It is important to use both verbal and written forms of communication and to allow patients to reflect over the information, ask questions, and be further informed over subsequent consultations. Consumer medicine information leaflets can be a useful aid, but require that the clinician understand and be prepared to answer questions raised by them. PMID- 14524207 TI - Nodular melanoma. No longer as simple as ABC. AB - BACKGROUND: Malignant melanoma is the fourth most commonly diagnosed malignancy in Australia. Nodular melanoma (NM) comprise less than 15% of all melanoma but account for up to 70% of those thicker than 3 mm. OBJECTIVE: This article describes the clinical features of NM, its prognosis and management. DISCUSSION: Nodular melanoma presents very differently from superficial spreading melanoma, and does not meet the 'ABCD' criteria used to alert doctors and patients to the possibility of this diagnosis. Due to their rapidly developing depth of invasion, urgent referral and wide excision are advised. PMID- 14524208 TI - Complaints and claims. What about you? PMID- 14524209 TI - Behavioural modification strategies for general practice. PMID- 14524210 TI - Body dysmorphic disorder--a fear of imagined ugliness. AB - BACKGROUND: Body dysmorphic disorder (BDD) is a distressing and impairing illness characterised by an intense time consuming preoccupation with an imagined or slight defect in appearance of a body part or parts. OBJECTIVE: This article outlines the characteristic features and treatment approaches for BDD and suggests screening techniques that can be usefully applied in the general practice setting. DISCUSSION: Body dysmorphic disorder has been shown to be common and chronic yet goes under recognised as the patient is often secretive about their distress. In an endeavour to alleviate the distress, time consuming rituals are performed that are either without effect or paradoxically may exacerbate the distress. Recent treatment studies show that the selective serotonin reuptake inhibitors and cognitive behavioural therapy, used simultaneously or alone, can be beneficial in alleviating the distress and impairment associated with the condition. PMID- 14524211 TI - Does high blood glucose mean more insulin? Type 1 diabetes management in children and adolescents. AB - BACKGROUND: The growth and development occurring in children and adolescents with type 1 diabetes contributes to many medical and nonmedical factors that may affect diabetic control. OBJECTIVE: This article discusses the assessment of high blood sugar levels in children and adolescents with type 1 diabetes. DISCUSSION: Traditionally, diet, exercise and insulin dose are seen as the determinants of blood glucose levels in type 1 diabetic patients. While these factors are important, other practical, medical and psychosocial factors need to be considered. Appropriate management requires more than just alteration of insulin dose. Insulin injection technique, adherence to insulin and management regimens in general, psychosocial issues, the role of intercurrent infections and the development of other medical problems need to be considered. Children and adolescents may only be seen by specialist physicians at three monthly intervals. Exploring these issues with patients during routine general practitioner consultations is likely to allow early identification of treatable problems and improve long term glucose control. PMID- 14524212 TI - Undertaking a literature search (with the patient present). PMID- 14524213 TI - New medical standards for drivers. AB - BACKGROUND: The national standards for assessing private and commercial vehicle drivers have recently been revised and combined into a single publication 'Assessing Fitness to Drive', published by Austroads. The new publication provides greater clarity of medical criteria for general practitioners and specialists and also details useful management guidelines. OBJECTIVE: This article introduces GPs to the new standards, outlines the key changes and implications for practice and highlights several medical, legal and ethical issues. DISCUSSION: Assessment of fitness to drive is a common issue in patient management and one that carries significant health, safety and lifestyle consequences both for the drivers themselves and other road users. PMID- 14524214 TI - Assessing fitness to drive. PMID- 14524215 TI - To freeze or not to freeze? Answering a question about the treatment of cutaneous warts. PMID- 14524216 TI - Patient with a leg ulcer. PMID- 14524217 TI - Corridor teaching. 'Have you got a minute...?'. PMID- 14524218 TI - Sterile corneal infiltrates. Contact lens case study--eye series 8. PMID- 14524219 TI - The development of family health nurses and family nurse practitioners in remote and rural Australia. AB - BACKGROUND: The World Health Organisation HEALTH21 strategy has firmly placed families and family oriented services at the core of health care delivery. OBJECTIVE: In this article we argue that a fundamental reorganisation of primary health care practices in remote and rural Australia needs to be undertaken. DISCUSSION: Nurses have been shown to be equally effective and less costly than general practitioners. Family nurse practitioners should be a first point of contact, and family health nurses should be responsible for responding to problems of multiple deprivation and social exclusion in remote and rural areas. These practitioners would, in their respective areas of responsibility, identify, diagnose, refer or treat individuals, families and communities. In effect they would act as gatekeepers to health and social care services. Family health nurses would also aim to support the community in developing and sustaining the capacity to take responsibility for its own health and social care. The main obstacles to these initiatives are concerns of general practitioners and nurses, the inertia of large organisations when faced with the need to undertake radical change and the highly unionised and rule bound nature of Australian nursing. PMID- 14524220 TI - Negotiating the future. The development of family health nurses and family nurse practitioners in remote and rural Australia. PMID- 14524221 TI - The Angolan civil war. A civilian disaster. PMID- 14524222 TI - The health consequences of child sexual abuse and partner abuse for women attending general practice. PMID- 14524223 TI - The impact of a history of child sexual assault on women's decisions and experiences of cervical screening. PMID- 14524224 TI - Training GPs in parent consultation skills. An evaluation of training for the Triple P-Positive Parenting Program. AB - BACKGROUND: The Triple P-Positive Parenting Program is a behavioural family intervention program that aims to prevent severe behavioral, emotional and developmental problems in children by enhancing the knowledge, skills, and confidence of parents. OBJECTIVE: This study evaluated the effect of training general practitioners in the use of the primary care version of the TPP on their consultation skills, satisfaction and confidence in conducting consultations with parents. STUDY DESIGN: Participants were assigned to an experimental condition that involved a brief behaviorally oriented parent consultation skills training program or to a wait-list comparison group. Thirty-two GPs participated in the training. Fifteen participants attended the first workshop (intervention group) and 17 attended the second (wait-list comparison group). RESULTS: GPs who participated in the training reported greater satisfaction with the outcomes of their parent consultations and showed significantly greater use of targeted parent consultation skills than GPs in the wait-list comparison group. Observations of GP consultation skills during simulated patient interviews with parents showed there was a significant overall improvement in their interactional skills during parent consultations. There was a high level of satisfaction with the quality of training received by the GPs. CONCLUSION: This was a brief, cost effective program that had significant effects on participating GPs' skills, confidence and satisfaction with child consultations involving behavioral problems. Implications for public health approaches to the prevention of child psychopathology are discussed. PMID- 14524225 TI - [It is normal for classification approaches to be diverse]. AB - It is asserted that the postmodern concept of science, unlike the classical ideal, presumes necessary existence of various classification approaches (schools) in taxonomy, each corresponding to a particular aspect of consideration of the "taxic reality". They are set up by diversity of initial epistemological and ontological backgrounds which fix in a certain way a) fragments of that reality allowable for investigation, and b) allowable methods of exploration of the fragments being fixed. It makes it possible to define a taxonomic school as a unity of the above backgrounds together with consideration aspect delimited by them. Two extreme positions of these backgrounds could be recognized in recent taxonomic thought. One of them follows the scholastic tradition of elaboration of a formal and, hence, universal classificatory method ("new typology", numerical phenetics, pattern cladistics). Another one asserts dependence of classificatory approach on the judgment of the nature of taxic reality (natural philosophy, evolutionary schools of taxonomy). Some arguments are put forward in favor of significant impact of evolutionary thinking onto the theory of modern taxonomy. This impact is manifested by the correspondence principle which makes classificatory algorithms (and hence resulting classifications) depending onto initial assumptions about causes of taxic diversity. It is asserted that criteria of "quality" of both classifications proper and classificatory methods can be correctly formulated within the framework of a particular consideration aspect only. For any group of organisms, several particular classifications are rightful to exist, each corresponding to a particular consideration aspect. These classifications could not be arranged along the "better-worse" scale, as they reflect different fragments of the taxic reality. Their mutual interpretation depends on degree of compatibility of background assumptions and of the tasks being resolved. Extensionally, classifications are compatible as much as they coincide by context and hierarchical structure of included taxa. Intentionally, typological classifications are compatible if included taxa are comparable by their diagnoses, while phylogenetic classifications are compatible if the included taxa are ascribed monophyletic status. A brief consideration is given to the "new phylogenetics" (= "genophyletics") as to a classificatory approach aimed at elaboration of parsimonious phylogenetic hypotheses based on molecular biology data and employing numerical methods of cladistic analysis. This approach is shown to borrows some phenetic ideas and revives scholastic principle of unified classificatory basis. It is supposed that, in a time, biological classification would get escaping from plethora of positivistic ideas (including those being developed by nowaday cladistics) and would assimilate (revive) more actively holistic worldview. PMID- 14524226 TI - [Pseudoreplication in ecological research: the problem overlooked by Russian scientists]. AB - The use of differential statistics to test for treatment effect with data from experiments where either treatments were not replicated (though samples may be) or replicates are not statistically independent leads to serious methodological problem. This problem, discovered by Hurbert (1984), is called pseudoreplication. Due to unknown reasons, pseudoreplication issue was completely overlooked by the Russian ecologists, in spite of the fact that the international scientific community is aware of pseudoreplication for almost twenty years. As the result, up to 47% of the experimental ecological papers, published in six Russian academic journals (Botanicheskij zhurnal, Ekologia, Izvestija RAN Ser. Biol., Lesovedenie, Zhurnal Obshchei Biologii, Zooligicheskij zhurnal) in 1998-2001, are pseudoreplicated; this proportion is nearly twice as high as the proportion of pseudoreplicated studies in international journals during 1960-1980, e.g. before the problem was discovered by Hurlbert (1984). This situation is alarming, especially because a substantial part of pseudoreplication arise from incorrect use of statistics, not from incorrect designing of experiments. By using several examples from the recent papers of Russian ecologists I shortly review the situations where pseudoreplication may occur and discuss some aspects of the experimental design, which are critical for correct processing and interpretation of ecological data. PMID- 14524227 TI - [Evolutionary saturation of species pool in present high mountain plant communities of Western Caucasus]. AB - The relative level of evolutionary saturation/nonsaturation of high mountain plant communities in Western Caucasus was defined by analysis of correlation between their local richness (LR) and size of species pool (SP). It is expected that if compared biotas are differently distant from the point of equilibrium one should look forward to a significant variation in correlation between LR and SP determined with the greater sensitivity of LR in comparison with SP in respect of abiotic factors. The lower is correlation LR/SP the greater is level of evolutionary nonsaturation in analysed communities. The results of analysis confirm the assumptions on 1. incompleteness of flora development in the high mountain zone in Western Caucasus, 2. different ages of alpine and subalpine communities, 3. different time necessary to attain the equilibrium with species pools of communities formed under extreme and more favourable conditions, 4. dependence of temps of flora development on the isolation of mountain ridges and massifs. The species pools of the most extreme alpine ecotope communities characterized by low environment capacity (plant groups of moving taluses, low species communities of alpine tapises, meadows and grounds) as well as the most of subalpine communities are evolutionary saturated or close to this condition. Development of species pools of alpine multi-species low-grass meadows as well as some of subalpine middle-grass meadows evidently is not completed yet. They can be considered to be the most probable places of the new species appearance by speciation or immigration from another Caucasus regions. PMID- 14524229 TI - [Computer simulation of different types of evolutionary process]. AB - The computer model of two alternative variants of biological evolution is proposed. The first variant supposes random while the second--directed change of individual features, thus corresponding to the Darwinian and non-Darwinian evolution. The evolution of fish communities in fresh waters serves as a particular example. The model is executed using object-oriented method of programming and mathematical apparatus of fuzzy logics. The investigation of the model showed that process of Darwinian evolution is connected with significantly greater species diversity and variability of evolutionary process trajectories than non-Darwinian one. On the other hand, non-Darwinian type of evolution provides fast achievement of high individual fitness, especially under conditions of constant environment. Non-Darwinian type evolution failed in big evolutionary alteration (for example, transition to predation); while the Darwinian evolution under the same conditions can produce such alterations though it took more time and many extinct species. Phylogenetic tree of Darwinian evolution is always more complex than of non-Darwinian one under the same conditions. PMID- 14524228 TI - [Hypophysis: a new scheme of ontogenetic development]. AB - It is usually accepted that hypophysis originates from two ecotodermal primordiums that give rise to its two main parts. The transformation of initial primordiums into definitive hypophysis is regarded as a result of mutual inductive influences between hypothalamus and Rathke's pouch. This scheme was remaining unchanged during many decades. Based upon the arguments that Spemann's organizer, prechordal mesoderm (PCM) and hypophysis are consecutive derivatives that replace each other during ontogeny, the author proposes that the two novel stages preceding the former starting point should be added to traditional scheme of pituitary development. Besides, according to new scheme the PCM (successor of Spemann's organizer) plays the main role in hypophysis development by active stimulation of morphogenesis in adjacent tissues. This leads to formation of Rathke's pouch and infundibulum, to their approaching each other, and to reprogrammin of Rathke's pouch ectoderm into hormonal cells of adenohypophysis. According to new hypothesis role of Rathke's pouch is accessorial and consists in provision of cell material. PMID- 14524230 TI - [The economical and evolutionary aspects of the optimal catch of the fish population]. AB - The natural interpretation of population value (internal cost) is used for the common mathematical problem of population exploitation. It is suggested that the population owner--the state--can use internal costs as a tax on fish caught by holders (fishermen). It turns out that such a tax outline make fisherman to establish an optimal long-term strategy of catch. Moreover, if there is several fishermen' the special tax, which makes them to be consistent with common cooperative strategy, can be worked out. According to the proposed hypothesis the changes in internal costs can be used as an adaptive response to the "demand and supply" deformation of the exploited population. The concept of "ecological economical" niche (habitat + place of sale) was proposed to characterize the exploited population. Computer calculations revealed the specific variant of Gause principle: the co-existing of two similar populations is impossible within single "ecologic-economical" niche. On the contrary, exploited similar populations show co-existence while fish is sailed on different markets. PMID- 14524231 TI - [The priority contribution of academician A.L. Kursanov's scientific school to the study of oxygen metabolism and anaerobiosis in plants]. AB - The biographical data on scientific activity of Academician A.L. Kursanov are presented. The contribution of A.L. Kursanov and his school to study of oxygen metabolism and plants anaerobiosis is reviewed. Special attention is given to the works on plant respiration, biosynthesis of endogenous water within plants and animals, mechanism of vitamin A biosynthesis and the metabolism of intracellular water studied with 18O methods. The important works of A.L. Kursanov's school that played an essential role in establishing the new brunch of science at the interface of physiology, biochemistry and ecology of plants, focused on the plant anaerobiosis theory are considered in details. PMID- 14524232 TI - [A problem of truth in biological systematics]. AB - A possibility to put a question of truth of knowledge in biological systematics is studied. It is shown that the problem of truth in reference to systematics is wider than a question of classified information reliability. Prerequisites needed for logically accurate formulation of a definition and criteria of truth are considered. It is shown that such prerequisites are present in taxonomic practice, namely in a process of diagnosis compiling. Philosophical analysis of this work has been carried out. Interpretation of an essence of systematics as classification is connected with use of classical concept of truth (which defines truth as correspondence between knowledge and object) in its undeveloped form. Carried analysis allows supposing that a theory of systematics based on diagnostics rather than on classification would be more prospective. Use of imperfect concept of truth can be seen also in notions that system of taxa must reflect its evolutionary history. Development and modernization of Aristotle's orientation to discovery of the object form can become an alternative to such opinions. An aspiration to achieve the truth is the main motive of systematic work. An influence of this aspiration on a selection of purposes of taxonomic work and theoretical comprehension of its bases is shown. Such features of modern biological systematics as its accessibility for new results, criticism in respect of external morphological characters, and interest in intraspecific variability are connected with this aspiration. This motive comes into contradiction with a tendency to withdraw the problem of truth as such, which takes place in some brunches of theoretic systematics. PMID- 14524233 TI - [Is "a personal computer and IT system" in the medical service rose and wine?]. PMID- 14524234 TI - [Pathophysiology of gastroesophageal reflux disease: motility factors]. PMID- 14524235 TI - [Esophageal motor function in nonachalasia motility disorders]. PMID- 14524236 TI - [A case of early carcinoma in tubulo-villous adenoma of duodenal bulbus]. PMID- 14524237 TI - [An autopsy case of sepsis due to Aeromonas hydrophila with hepatic cirrhosis type B]. PMID- 14524238 TI - [A case of port-systemic shunt associated with splenic artery aneurysm and pulmonary hypertension]. PMID- 14524240 TI - [A case of Caroli's disease which DIC-CT was useful to a definite diagnosis]. PMID- 14524239 TI - [A case of hepatocellular carcinoma treated with TS-1]. PMID- 14524241 TI - [A case of autoimmune pancreatitis with Harada's disease]. PMID- 14524242 TI - [A case report of an advanced esophageal carcinoma (stage II) without signs of recurrence in 5 years after chemoradiotherapy. With reference to p53 immunohistochemistry as an indicator for prognostic evaluation of chemoradiotherapy]. PMID- 14524243 TI - [Study on virulence factors of bacterial enteropathogens, especially Vibrio spp. and enteropathogenic E. coli, and the application study]. PMID- 14524244 TI - [Bacillus anthracis and anthrax]. PMID- 14524245 TI - [Therapeutic strategies in cases of septicemia caused by gram-negative bacteria]. AB - Septic shock is the most severe manifestation of infection and appears to be increasingly common, especially in the intensive care units. Lipopolysaccharides (LPS, endotoxin) are known to be responsible for the initiation of septic shock, therefore they can be targets for new preventive and therapeutic strategies. This review focuses on endotoxin-based molecular strategies for the prevention and treatment of Gram-negative sepsis and septic shock. PMID- 14524246 TI - [Bronchial asthma as a multifactorial disease]. AB - Asthma is a common condition that results from the interaction of an unknown number of genes with environmental factors. Here, we describe the recent advances in this field including results of genome-wide searches for susceptibility genes. PMID- 14524247 TI - [Endocytosis mediated by receptors--function and participation in oral drug delivery]. AB - The rapid expansion of applied biotechnology research in current pharmaceutical drug discovery has resulted in the development of increasing numbers of novel macromolecular therapeutics. Oral bioavailability of these compounds is usually poor due to a combination of incompatible physicochemical properties, resulting in low cellular penetration, and high susceptibility to metabolic enzymes present within the gastrointestinal tract. For chronic drug therapy, however, the oral pathway is generally considered to be the most convenient route of administration. Thus far, limited success has been observed in oral macromolecular delivery by either concomitant administration of penetration enhances or the use of prodrug strategies involving carrier-mediated transport systems. An alternative approach may be targeting to receptor-mediated endocytosis (RME) systems. It is the aim of this mini-review to present a concise overview of the latest developments in RME research and RME systems that are either currently targeted for the oral delivery of macromolecules or may appear to become potential targets for future oral drug delivery strategies. PMID- 14524249 TI - [Lipoprotein (a)--structure, properties, physiologic and pathologic significance]. AB - The data on the structure and the role of lipoprotein (a) were presented. It posses plasminogene and lipoproteins elements in its structure. Up to now it is not well known the physiological role of lipoprotein (a). There are many proofs in the literature that lipoprotein (a) is an independent coronary risk factor. It is also known as an acute phase protein. PMID- 14524248 TI - [Dopamine receptors--structure, characterization and function]. AB - Dopamine (DA) is an important neurotransmitter/neuromodulator that plays various functions within the body. In the central nervous system, DA is involved in the control of locomotion, cognition, emotion, neuroendocrine secretion and function of retinal cells. In the periphery, DA participates in the regulation of homeostasis, vascular tone and hormone secretion. The diverse physiological functions of DA are mediated by at least five distinct membrane bound receptors, i.e. D1 and D5--members of D1 family, and D2, D3, D4--members of D2 family of DA receptors. All DA receptors belong to the superfamily of G-protein-coupled receptors. This survey summarizes current knowledge on the molecular and pharmacological characterization of DA receptors, their role in the regulation of various processes in an organism, and involvement in the therapy of several disorders, in particular in the treatment of Parkinson's disease, schizophrenia, and hypertension. PMID- 14524251 TI - [Progress in understanding genetics of allergy and asthma]. AB - Progress in genetics of allergic diseases is based upon the basic science development. Some problems in genetic predisposition to disease and prognosis of clinical course have been discussed and presented in relation to allergic diseases. Moreover, pharmacogenetics of asthma and allergy have been pointed in relation to beta 2-adrenoreceptor. PMID- 14524250 TI - [The role of caspases in the apoptosis process]. AB - Activation of caspases is the key event during apoptosis. Abnormalities of this phenomenon play an important role in the pathogenesis of several disorders and may have important clinical implications, including the development of novel therapeutic strategies. Currently, these problems are extensively investigated in several experimental and clinical studies. PMID- 14524252 TI - [Genetic studies in allergology]. AB - A wide application of molecular biology methods provided the basis for genetic studies of complex diseases, like allergic diseases. Analysis of genetic predisposition for these diseases should be performed at the level of individual phenotype, atopic background and differences in severity of individual diseases. The most frequently used methods of identification of disease-causing genes include positional cloning and candidate gene studies. In spite of huge advances in genetic studies the genetic predisposition for allergic diseases has not been fully elucidated. PMID- 14524253 TI - [The role of radical oxygen species in airway inflammation]. AB - Reactive oxygen species (ROS) are among numerous mediators of airway inflammation. They are playing a key role in asthma, chronic obturative pulmonary disease (COPD), adult respiratory distress syndrome (ARDS) and lung fibrosis (LF) pathogenesis. In view of very short half-life, their effects are local, related to the site of their formation. Generated either by epithelial or infiltrating cells, ROS cause epithelial damage, increase vascular leakage, induce smooth muscle proliferation and enhance pro-inflammatory gene expression. They also increase arachidonic acid mediator release through cytosolic phospholipase A2 phosphorylation. Drugs inhibiting ROS generation have become the first line of treatment of airway inflammatory disorders. PMID- 14524254 TI - [Cytokines in allergic inflammation]. AB - Cytokines play a central role in the pathogenesis of allergic diseases and allergic inflammation. Therefore, an understanding of mechanisms which regulate production and function of cytokines is very important and may result in the development of more effective methods of treatment of allergic diseases. Recent studies have demonstrated that the induction of allergic inflammation requires genetic background and environmentak factors. The most important cytokines and chemokines are IL-4, IL-5, IL-3, IL-13, GM-CSF and TNF-alpha. Many other cytokines are responsible for the growth, maturity, migration activation and apoptosis of all cells involved in allergic inflammation, among them are IL-2, IL 5, IL-6, IL-9, MIP-1 alpha, RANTES, IL-8, IL-12, IL-18, IFN-alpha i gamma, TGF beta, sIL-4, IL-1Ra. Recently it has been proven that IL-10 and other cytokines from the IL-10 family, and TGF-beta have anti-inflammatory properties in allergy. PMID- 14524255 TI - [The importance of inhibition of adhesion molecules in allergic inflammation]. AB - Inflammation is a defensive reaction of an organism in response to injuring factors and is characterised by effector cells infiltration. Adhesion molecules are involved in their ordered influx. These molecules are glycoprotein particles present on cell surface and on intracellular matrix proteins. Cytokines, chemokines and other inflammatory process mediators modulate the expression of adhesion molecules. Anti-adhesion therapy involves many techniques. Most frequently monoclonal antibodies are used. Other forms of therapy contribute to blocking the synthesis of molecules at transcription level or to inhibition of their transfer from cell interior to its surface. Soluble molecule forms or their receptors are also applied. PMID- 14524256 TI - Changing society, changing allergens. AB - Changes in the political system in Lithuania have been reflected in different allergens. In the paper some changes in the house dust fauna and in the air are presented in relation to the new situation and in comparison to the previous period. PMID- 14524257 TI - [Pollen monitoring in Poland]. AB - The author presents the history of aerobiological studies in Poland. The first scientist to undertake research regarding this problem in Poland was Mieczyslaw Obtulowicz. The research on this topic was resumed again in Krakow and Rabka in the 1960s. since 1989, constant measurements of plant pollen and fungal spore concentrations have been conducted by volumetric methods in over ten sites in Poland, in a network created by P. Rapiejko. In 1989 the first pollen report, worked out by the Allergen Research Centre appeared in daily newspapers. Pollen information (current data reports and forecast) is available to the public through radio broadcasts, television and newspapers. Pollen calendar for Poland is presented. Pollen sampling methods employed in these studies are also briefly described. PMID- 14524258 TI - [The impact of environmental factors in atopic disorders]. AB - The widely understood environmental influence plays an important role in the formation of atopic phenotype. The manifestation of clinical symptoms and signs of asthma is the resultant of the effects of many genes and gene-gene and gene environment interactions. Based on recent literature data the main environmental factors are analysed, such as tobacco smoke, viral infections, living conditions, moisture and moulds, diet and air pollution. Understanding of these genetic determinants and the role of environmental factors would make possible to improve the methods of diagnosis and treatment. PMID- 14524259 TI - [Importance of atypical infection in asthma]. AB - The category of atypical respiratory infection involves viruses, bacteria and rickettsia. These micro-organisms can play a marked role in the exacerbation an asthma course but also participate in its development. Authors discuses both aspects, reporting own observations. The knowledge of real importance of atypical infections in asthma remains to be established. PMID- 14524260 TI - [Contemporary views on the pathological mechanism of asthma]. AB - Asthma is a chronic inflammatory disease of the airways. Genetic and environmental factors contribute to the development of asthma. Both inflammation and remodelling are essential mechanisms in the development of subepithelial fibrosis and smooth muscle hypertrophy leading to progressing decrease of FEV1. Inflammatory changes in asthma are characterised by cellular infiltration of airway epithelium, bronchial wall and smooth muscle layer. The most important cells participating in asthma include T lymphocytes, mast cells, airway epithelial cells, eosinophils, antigen-presenting cells, neutrophils, fibroblasts, myofibroblasts and macrophages. They produce many cytokines responsible for the development of allergen-specific clones of Th2 lymphocytes, for production of IgE by B-cells, for increased expression of adhesion molecules, for migration of cells, their activation and release of inflammatory mediators. The mediators coming mainly from mast cells and eosinophils that include leukotrienes, prostaglandins, histamine, alkaline proteins and enzymes are responsible for airway obturation that results from bronchospasm, bronchial mucosal oedema and excessive amount of abnormal mucous secretion. An important role in the obturation is played by smooth muscle hypertrophy and deposition of proteins under the basal membranes of respiratory epithelium. PMID- 14524261 TI - [The role of immunoregulatory disorders in pathogenesis of bronchial asthma]. AB - The set of cellular and humoral elements which participate in the pathogenetic chain of bronchial asthma is regarded as characteristic for this disease. They include, among many others: mast cells, basophils, eosinophils, macrophages, Th2 phenotype lymphocytes, IgE antibodies, histamine, leukotrienes and some pro inflammatory cytokines. The majority of them are effectory agents active at the end of pathogenic immune response. The therapy antagonizing their influences alleviates the symptoms but does not eliminate the cause of the illness. The authors call the attention to the pathogenic significance of immunoregulatory disorders which appear during the induction phase of immune response in patients with symptoms of bronchial asthma. On the basis of current literature and own, already published studies, the authors point to the significance of recently discovered and described thymus-dependent cellular population of regulatory T lymphocytes which are active in prevention of development of allergic and autoaggressive reactions. It is suggested that deficiency of regulatory T cell population caused by undesired environmental influence on thymic lymphopoiesis and the concomitant overstimulation of immunogenic antigen-presenting dendritic cells, create the conditions under which pathogenic immune response may well develop. Therefore, the possibility of introducing of prothymic immunocorrective treatment in bronchial asthma should be taken into account in further clinical studies. PMID- 14524262 TI - [Lung remodeling in asthma]. AB - Repair is an integral part of every inflammation. Injured cells are replaced by new cells of the same type or by connective tissue. This may result in restitution "ad integrum" or lead to structural changes referred to as remodelling. For many years the importance of this phenomenon for the course of asthma, an undoubtedly chronic inflammatory disease, was underestimated. The presence of only partial reversibility of airflow obstruction in some patients with asthma has significantly changed our view on that problem. The irreversible component of airway obstruction is more prominent in patients with severe disease and persists despite aggressive anti-inflammatory treatment. Recent studies have proved that remodelling in asthma encompasses not only the bronchial tree, but also alveolar tissue. There are many factors supposedly responsible for lung reconstruction characteristic of asthma: mediators, cytokines, and growth factors. Better understanding of these processes should provide a basis for novel, long-term therapeutic strategies in asthma. PMID- 14524263 TI - [Latent sources of contact allergy]. AB - In individuals sensitized to many contact allergens in the course of 4th immunological mechanism, in whom allergic contact dermatitis develops at the site of exposure to haptens, disseminated eczematous skin lesions might be provoked by latent source of hapten, which penetrates to the body omitting the skin. The most frequent clinical situations include: allergy to drugs, food additives, inhaled chemicals originated from plants, allergic reactions to metal endoprostheses used in orthopaedics and/or dentistry, and all conditions facilitating penetration of hapten directly to the blood through damaged skin (erosions, ulcers, etc.) and/or under occlusive dressings. PMID- 14524264 TI - [Allergic phenomena in the course of dermatomycoses]. AB - Fungi, well known as infectious factors so far, are important allergens too. A fungal id reaction is defined as a distant skin manifestation of an established fungal infection. The most typical setting is in association with tinea of the feet. The main id reactions are well established. A widespread lichen trichophyticus--eruption of small follicular papules grouped or diffuse. The eruption is symmetrical, usually pronounced on the trunk but in severe cases extending down to the lower limbs, even sometimes involving the face. The common cause of this type of id reaction is scalp ringworm of kerion type, caused typically by Trichophyton verrucosum. The treatment of the original ringworm lesion may play a role in initiation of the process. A pompholyx-like id affecting the interdigital spaces and palmar surfaces of the fingers, the palms and sometimes the dorsal surfaces of the hands. This eruption is associated with an acute inflammatory tinea of the feet and may develop spontaneously or as a result of inappropriate treatment. The palmar and web space skin may be covered with papules or vesicles. Sometimes blisters or pustules may occur. Other varieties of mycides include eczematous eruptions, erysipelas-like dermatitis, psoriasiform dermatitis, erythema multiforme, erythema annulare centrifugum, erythema nodosum, and urticaria. Adequate antifungal therapy is recommended as the treatment of allergic diseases caused by superficial fungi. PMID- 14524265 TI - [Influenza in global aspect]. AB - Due to a high ability to change, the influenza virus is still a danger for humans. The outbreak of the next influenza pandemic is very probable. The disturbing events that occurred in Hong Kong in 1997 and in the beginning of 2003 confirmed such a possibility. Every year in Poland influenza causes from several hundred thousands to several millions of cases of infections. All over the world the annual number of influenza cases amounts to approximately 100 millions and from 0.5 million to 1 million patients die due to influenza. The most severe threat is connected with post-influenza complications that are the cause of death after influenza infection. The only one effective method of prophylaxis against influenza and its complications is vaccination. Moreover, since 1999 new anti influenza drugs are available: oseltamivir (Tamiflu) and zanamivir (Relenza). They are effective for treatment or for treatment and prophylaxis of infections caused by influenza virus type A and also type B. The efficacy of these drugs depends on appropriate diagnosis. Due to this fact laboratory diagnostics of respiratory infections is so important and crucial. It should be emphasised that clinical symptoms of infection may indicate influenza even if the disease is caused by other viral or bacterial pathogen. PMID- 14524266 TI - [Diagnosis of allergic diseases]. AB - This study presents the general rules of diagnosing allergic diseases, which can be reduced to answers for the following three questions: is the patient allergic? (does he/she belong to the group of people at risk for allergy?). If so, then; what is he/she allergic to? And again, if the answer is positive, then: does the substance he/she is allergic to, gives the reasons for the symptoms? As it follows from the above-mentioned questions, besides determining the disease, the main goal of allergy diagnostics is to recognize the allergen responsible for the symptoms. In order to attain this, several widely recognized tests are performed (skin prick tests, allergen specific IgE, skin patch tests, allergen challenge tests, etc.). Some doctors use the testing methods which have not been scientifically proven to be effective and accurate. As a matter of fact, many patients in Poland are tested with unverified (ALCAT) or negatively verified methods (allergen specific IgG4) as well as by means of alternative medicine (acupuncture, BICOM). PMID- 14524268 TI - [Specific nasal challenge tests in the diagnosis of allergy]. AB - The paper presents a review of current literature concerning the use and conduction of specific nasal challenge tests (SNCT). The authors call attention to the consensus of the German group of 1990 and report of the European Rhinologic Society (ERS) on nasal challenge tests of 2000. They present own experience and point to the unsolved questions: whether SNCT should be one- or two-sided with allergen on blotting paper or from a spray, and what methods of subjective and objective assessment should be applied. The authors call attention also to the unsolved problem of the influence of nasal cycle on the challenge test result. The paper includes the ERS instruction how to conduct nasal challenge test. PMID- 14524267 TI - [Bronchoalveolar lavage and induced sputum in asthmatic and COPD patient]. AB - Scientific studies of the material obtained during bronchial biopsy, bronchoalveolar lavage and sputum induction altered the view on the pathogenesis of asthma and COPD. In the result, both bronchial diseases are considered inflammatory. Eosinophils play the main role in the pathogenesis of asthma, whereas in COPD the most significant are neutrophils. Bronchial biopsy and bronchoalveolar lavage have been applied mainly in scientific studies, while sputum induction as a non-invasive method can be useful in everyday practice too. Increased level of eosinophils in sputum allows to forecast the efficiency of glucocorticosteroids in the therapy of asthma and COPD. PMID- 14524269 TI - [Diagnosis and treatment of urticaria and angioedema]. AB - Current concepts on the diagnosis and treatment of urticaria and angioedema concerning allergic, non-allergic, idiopathic and symptomatic aetiology of this very complex disease were discussed. Much attention was paid to the necessity of multidrug therapy in which antihistamines play the most important role in chronic forms of the disease. The possibility of occurrence of life-threatening conditions was emphasised and the rules of their treatment and early prevention of intensification of symptoms typical of anaphylaxis. PMID- 14524270 TI - [Diagnostic problem of ordinary chronic urticaria]. AB - On the basis of literature data new classification of chronic urticaria was presented, depending on pathogenetic factors and clinical picture. The most frequent causes of the disease, such as food, drugs, infectious and hormonal factors were discussed in detail. The relationship between ordinary chronic urticaria and other systemic diseases was underlined. PMID- 14524271 TI - Ethical issues and problems in clinical research. AB - In the paper some documents on clinical research have been presented. Declaration of Helsinki WHO documents are suggested as a basic papers in this field. Some "delicate" problems are pointed and discussed in question about placebo and "moral obligations". PMID- 14524272 TI - [Asthma with hypersensitivity to aspirin]. AB - Aspirin-induced asthma (AIA) is a distinct clinical entity characterised by hypersensitivity to aspirin and other non-steroidal anti-inflammatory drugs (NSAIDs) typically occurring in the form of asthma and severe rhinosinusitis. Its prevalence is about 10%, but AIA probably is underdiagnosed. The diagnosis can only be established with certainty by challenge tests, using increasing doses of aspirin. The crucial cell in AIA is the eosinophil, while the role of mast cell is more elusive. At the biochemical level, AIA is distinguished by profound underlying arachidonic acid disturbances. An overproduction of leukotrienes was observed in patients with aspirin hypersensitivity at baseline and following aspirin challenge. In some patients altered prostaglandin release following aspirin was encountered. The inhibition of COX-1, but not COX-2, was shown to precipitate post-challenge symptoms precipitated by aspirin. The new insights into eicosanoid molecular biology and genetics have recently emerged. The patients with aspirin-sensitive asthma require often prolonged oral, inhaled and nasal corticotherapy. Antileukotrienes also can be used. Aspirin and NSAIDs should be avoided. Nevertheless, highly specific COX-2 inhibitors are well tolerated. Aspirin desensitisation, followed by daily aspirin treatment, is a valuable therapeutic option. PMID- 14524273 TI - [Bronchial asthma infectious exacerbations]. AB - Bronchial asthma and asthma-like form of COPD often undergo exacerbations with symptoms of infection. Currently, there is a general agreement, that most of these infections that exacerbate asthma especially in children are caused by viruses. Several "common cold" viruses are known to cause these exacerbations (RSV, parainfluenza virus, rhinoviruses). To date, there is no certainty, if viruses exacerbate asthma alone or in combination with allergen. It is also unknown, whether they can induce primary asthma symptoms. There is a growing support for the opinion, that in infectious exacerbations of asthma also bacteria can play an important role. These include both typical as well as atypical bacterial strains (Chlamydia pneumoniae). The effectiveness of antibiotics and bacterial immunotherapy with vaccines seems to support the role of bacteria in asthma exacerbations. Another very important problem is related to the role of infectious agents in prevention of atopy. Many facts have been gathered supporting the so called "hygienic theory". PMID- 14524274 TI - [Influence of influenza virus infection on asthma and COPD]. AB - The recent results of clinical trials and evaluations of connections between viral infection, especially influenza virus infection, and clinical course of bronchial asthma and COPD were presented in the paper. Special attention has been paid to real efficacy of influenza vaccinations in risk groups and possibilities of clinical control. PMID- 14524275 TI - [Rhinitis and sinusitis: influence on asthma and COPD]. AB - Basing on epidemiology, pathophysiology and clinical manifestations we can show an important correlation between rhinitis and bronchial asthma. Prevalence of allergic rhinitis in childhood is an early manifestation of atopy. It is estimated that allergic rhinitis occurs in about 60% of patients before the onset or simultaneously with bronchial asthma. The epidemiologic, etiopathogenetic and clinical data demonstrate that the upper and lower airways are one system, which, in the case of allergic inflammation induces common mechanisms intensifying and maintaining this process. There is no doubt that allergic rhinitis is a risk factor of asthma development. Early diagnosis, treatment and prophylaxis should involve both the upper and lower airways. Coexistence of sinusitis and nasal polyps is another factor causing bronchial hyperreactivity and asthma symptoms. PMID- 14524276 TI - [Organic dust-induced allergic respiratory diseases]. AB - Epidemiology, pathogenesis, clinical picture, diagnosis, management, and prognosis of allergic alveolitis are described in this paper. PMID- 14524277 TI - [Allergy of the oral cavity, throat and larynx]. AB - Current views, mainly clinical, concerning allergic inflammation in the upper respiratory tract are presented. PMID- 14524278 TI - [Allergy and hypersensitivity to drugs: anaphylaxis]. AB - Anaphylaxis due to allergic or non-allergic hypersensitivity to drugs is an acute fatal or potentially fatal reaction. The estimate rate of 3.2 cases of drug associated fatal anaphylaxis per 100.000 inhabitants per year seems to be approximately 10 times greater in hospitalised patients. The most common agents that are responsible for anaphylaxis in hospitalised patients are muscle relaxants, latex, antibiotics, anaesthesia-inducing drugs, plasma expanders followed by local anaesthetics and iodinated radio contrast media, and many others. Antibiotics and non-steroid anti-inflammatory drugs (NSAID) are the common cause of anaphylaxis in non-hospitalised patients. Predictive tests for high-risk patients have not been established. However, several procedures for prevention are proposed. These include preoperative screening to detect sensitisation to anaesthetic drugs and latex, or pre-treatment with antihistamines and corticosteroids to prevent anaphylaxis to iodinated contrast media. Challenge tests are useful for the determination of a safe anaesthetic or NSAID. Unfortunately, the causative agents cannot be identified in all cases of anaphylaxis. Subjects at risk for idiopathic anaphylaxis should have epinephrine available for self-administration. PMID- 14524279 TI - [Drug-induced skin allergy]. AB - Drug allergy is an important and increasing problem in everyday practice. It is estimated that about 15% of adverse side effects of drugs are of allergic nature. This kind of adverse reactions appear where antibodies or activated T cells are directed against drugs. The pathomechanism and symptoms of drug allergy may be connected with every of 4 types of hypersensitivity according to Gell and Coombs. Most frequently, drug allergy manifests as different forms of skin reactions. Some of them like urticaria and angioedema are IgE-related and appear rapidly after drug intake. Others appear as delayed-type reactions and manifest as maculopapular, pustular or bullous exanthema. The diagnosis of drug-induced skin allergy is based on history, clinical picture, in vitro laboratory tests and skin tests (prick, intradermal and patch). The treatment includes drug withdrawal and symptomatic pharmacotherapy. PMID- 14524280 TI - [Skin adverse drug reactions]. AB - Adverse drug reactions are common and important complications of therapy. The clinical picture of drug-induced reactions is characterised by considerable polymorphism: from mild local lesions to fatal systemic processes. The classification of drug hypersensitivity encompasses systemic or specific organ forms. Skin has a special position in the diagnostics of adverse drug-induced reactions. Cutaneous lesions are almost universal and highly pathognomonic symptoms of drug-induced hypersensitivity. Urticaria, erythema-like skin reactions, drug-induced exanthemas, purpuric and hemorrhagic reactions are the most common cutaneous drug-induced manifestations. PMID- 14524281 TI - Atopic dermatitis as an actual problem of modern allergology. AB - The etiology of atopic dermatitis is multifactorial and therefore therapy design of this disease should be complex and should cover all sites of pathologic process that underlies this disease. The therapy of atopic dermatitis is presented according to actual state. The importance of topical therapy is held considering patient's age, stage of the disease, dispersion and severity of the clinical manifestation, effect and tolerance of the previous treatments. PMID- 14524282 TI - [Allergic contact dermatitis]. AB - Clinical definition of eczema is based on the presence of multiform erythematous, papular and vesicular lesions which are followed by marked desquamation. When the underlying mechanism is allergic, skin lesions are mediated by inflammatory Th1 lymphocytes recognizing hapten determinants, i.e. IVth allergic mechanism in allergic contact dermatitis. This mechanism is responsible also for allergy to bacterial proteins in nummular eczema, or to fungal and bacterial proteins, atopens and nickel in dyshidrotic eczema. Eczematous lesions can develop after damage to the skin barrier by toxic chemical or irritating substances, as non allergic irritation eczema. These substances non-specifically stimulate the Langerhans cells and keratinocytes to produce numerous cytokines inducing expression of adhesion molecules on the endothelial cells. This is responsible for non-specific mobilization of Th1 cells to the skin. A model example is dermatitis induced by mercury salts and/or sodium lauryl sulfate. Intermediate position between allergic and non-allergic eczema is occupied by atopic dermatitis/eczema. In addition to classical IgE-dependent mechanism leading to degranulation of mast cells, there is generation of specific population of Th2 lymphocytes recognizing food and air-borne atopens. This reaction is responsible for skin inflammation in the late-phase response, in which allergic process started by Th2 cells switches to non-specific migration of Th1 cells attracted by cytokines released from eosinophils, keratinocytes and Th2 lymphocytes. PMID- 14524283 TI - [The new possibilities to control allergic inflammatory processes]. AB - The development of immune or allergic reactions is involved in the induction of inflammation. The apoptotic elimination of responsible cells prevents or diminishes inflammation. In 2002 S. Brenner, J. Sulston and R. Horovitz were awarded the Nobel Prize for the discovery and description of the mechanisms and role of the preset cell death, which already in 1972 was called apoptosis by J. Kerr. Apoptosis plays an important role in growth and development and also maturation processes of the immune system. It is precisely regulated by appropriate genes. Autoreactive T cells are eliminated in the thymus by apoptosis. The depletion of regulatory T cells by deviated reactions of apoptosis may be responsible for the induction of autoimmunity of allergy in view of absence of peripheral tolerance or impairment of regulatory CD4+ and CD25+ function. Induced or directed apoptosis of certain cells may be a useful adjunct to the therapy of allergic diseases. PMID- 14524284 TI - [Cytokines and anti-cytokines in allergic diseases]. AB - Allergic inflamation is complexed phenomenon related to the activity of many mediators released from "effector cells". The role of IL-12, IL-5, IL-4 and some adhesive molecules is presented with special attention focused on therapeutical aspects in allergic diseases. PMID- 14524285 TI - [Anti-IgE therapy]. AB - Atopy--the genetic predisposition to produce allergen-specific IgE is one of the most important predisposing factors for the development of asthma and other allergic diseases. Because of pivotal place of IgE in early and late phase e of allergic responses the introduction of anti-IgE antibodies to therapy provides a novel and promising approach to treatment of asthma and other IgE-dependent diseases. The administration of anti-IgE reduces circulating IgE levels to less than 1% of the baseline value and down-regulates the expression of basophil IgE receptors. Anti-IgE antibody rhuMAb E25 (omalizumab) improved asthma symptoms, reduced the rates of exacerbations and allowed for the reduction of corticosteroids and rescue medication doses. Anti-IgE therapy is safe and well tolerated. PMID- 14524286 TI - [The possibility of monitoring of bronchial hyperresponsiveness]. AB - The aim of this review was to present the evidence of the role of the measurement of bronchial hyperresponsiveness in monitoring of asthma. Airway hyperresponsiveness can be assessed during regular clinic visit and it is related to the severity of asthma and airway inflammation. The indirect markers of airway hyperresponsiveness, such as AMP or hypertonic saline may more accurately reflect airway inflammation and remodelling than routine symptoms and lung function monitoring. Monitoring of patients with asthma is a key aspect of optimal asthma management and the measurement of airway hyperresponsiveness may improve asthma control. PMID- 14524287 TI - [Safety and efficacy of inhaled ciclesonide in asthma]. AB - Poor adherence to medications and other aspects of the treatment plan are described in patients with asthma, especially with adverse events concerning the upper respiratory and digestive tracts. Against this background a review is presented of current knowledge about the new class of glucocorticoids (ciclesonide). PMID- 14524288 TI - [The influence of inhaled glucocorticoids on the growth of asthmatic children]. AB - In this paper general characteristic of the growing process of children is described. Various factors may modify this process including genetic predisposition, environmental factors, diseases, diet, and drugs. Asthmatic children present growth retardation, however without significant influence on final height. Low and moderate doses of inhaled glycocorticosteroids have no significant effects on growth. The results of short, medium and long term studies are discussed. PMID- 14524290 TI - [Clinical aspects of anti-inflammatory action of antihistamines]. AB - Antihistamines are accepted in the therapy of allergic seasonal and perennial rhinitis. In the paper some anti-inflammatory effects of antihistamines have been presented, and their action mechanisms and clinical applications have been discussed in relation to the new preparates of antihistamines. PMID- 14524289 TI - [Anti-leukotrienes in allergic diseases in children]. AB - In the paper a review is presented of current literature on the use of antileukotrienes in the treatment of allergic diseases in children. Recent reports suggest a role of cysteinyl leukotrienes in the development of inflammatory reaction in the airways, including bronchial remodelling. PMID- 14524291 TI - [Antihistamine drugs in the treatment of allergic skin diseases]. AB - Antihistaminic therapy is widely used in the treatment of allergic dermatoses. Second generation antihistamines due to their pharmacokinetic mechanism depending on the wide extra-receptor anti-inflammatory and anti-allergic actions are used in the treatment of diseases, in which the main pathogenetic role is played by various allergic mechanisms and inflammatory cellular processes. The modern views on etiopathogenesis and antihistaminic treatment of allergic skin diseases are presented in this paper. The course of allergic inflammation, role of histamine and its receptors, taking into consideration the biological effects of their suppression and influence on the course of early and late phase of allergic reaction, are discussed. A review of classical and second generation antihistamines has been done, taking into account their usefulness in the treatment of acute and chronic urticaria, angioedema, atopic dermatitis, different forms of eczema and various drug-induced reactions. PMID- 14524292 TI - [Safety of new antihistamines]. AB - In this paper the new antihistamines: fexofenadine, desloratadine, norastemizole, levocetirizine etc. are presented. The toxicity and adverse effect of these drugs are discussed with special regard to interaction with other drugs and their influence on sedation and circulatory system. PMID- 14524293 TI - [Therapeutic management in severe and difficult asthma in adults]. AB - Classification of severity of bronchial asthma is a basic point for therapeutic management in adults. In the paper definitions of severe and difficult asthma are presented. The therapeutic possibilities are pointed according to new guidelines published in the recent period and own Author's clinical experience. PMID- 14524294 TI - [Usefulness of antibiotics in the treatment of bronchial asthma]. AB - The efficacy and safety of antibiotics in the treatment of asthma are not unequivocal. Among various antibiotics, macrolides play the main role in the therapy of bronchial asthma. Macrolides are useful in the treatment of asthma not only because of their antimicrobial activity. The mechanism of action of macrolides in improving asthma and reducing airway responsiveness is evaluated especially due to their activity in non-infectious asthma. Macrolides may not only enhance the host defense system through increased cytokine synthesis, but also exhibit anti-inflammatory activity mediated by anti-inflammatory cytokines. The steroid-sparing effect of macrolide antibiotics has been postulated to contribute to their beneficial actions in the treatment of asthma. Macrolides may be useful in the treatment of patients with steroid-dependent asthma, probably because they inhibit eosinophilic inflammation. It has also been suggested that the effect of macrolides on bronchial hyperresponsiveness is mediated by their inhibitory action on superoxide production and chemotaxis of polymorphonuclear neutrophils and the mixed lymphocyte reactions. In spite of these suggestions, the mechanism of action of macrolides in asthmatic syndrome is not clear. Only well-designed and conducted clinical studies are capable of assessing the efficacy and safety of immunosuppressive macrolides in the treatment of asthma. PMID- 14524295 TI - [The development of allergy to antibiotics]. AB - The paper includes a survey of issues concerning evolution of allergy to antibiotics. Particular attention is paid to penicillin allergy as this antibiotic has been the drug most frequently causing sensitization since 1960s until today, although the frequency of the allergy is continuously decreasing. In addition, the paper deals with the importance of replacing benzyl penicillin with semi-synthetic penicillins and the impact of these process on the nature of immune response (a decrease in the immediate allergy incidence, an increase in the delayed allergy incidence), cross reactions, the significance of the hidden allergen sources. Apart from that, the paper discusses the most important issues concerning the evolution of allergy to other antibiotics, especially the aminoglycoside antibiotics and cephalosporins. PMID- 14524296 TI - [How to obtain complete control of asthma?]. AB - Long acting beta 2-agonists (LABA) and glycocorticosteroids (GCS) are the most effective groups of drugs recommended in the treatment of asthma. They are more effective if administered together. GCS have been used as anti-inflammatory agents while LABA as bronchodilatatory drugs. It has not been fully elucidated whether LABA exert in vivo any anti-inflammatory effect that was demonstrated in vitro. The interaction between these drugs at the biochemical and clinical levels was confirmed many times. The mechanisms of interactions between GCS and LABA are, however, still being discussed. LABA may affect glukocorticosteroid receptor (GR) location through modulation of GR phosphorylation and further may prime GR function within the cell nucleus by modifying GR or GR-associated protein phosphorylation. Glucocoricosteroids may, in turn, regulate beta 2-adrenoceptor (beta 2 AR) function by increasing its expression, acting by restoring the ability of G-protein beta 2 AR coupling and inhibiting beta 2 AR down-regulation, thereby preventing desensitisation associated with long term LABA administration. PMID- 14524298 TI - [Diversities in the treatment of patients with chronic obstructive lung disease]. AB - Bronchial asthma and chronic obstructive lung disease are characterised by dyspnoea, cough and, during exacerbation, wheeze in some patients. The common pathophysiological element is obturation of the bronchial tree and chronic inflammation. These diseases differ, however, in location of the inflammatory process in the respiratory system, various degree of participation of inflammatory cells and inflammation mediators. These elements influence the effectiveness of treatment. In both diseases inhalant and systemic glucocorticosteroids, beta 2-mimetics, cholinolytics, theophylline. The reaction to these drugs is, however, different. In the paper, differences in treatment of these diseases are also discussed. PMID- 14524297 TI - [Recent results of clinical studies of synergistic treatment in obstructive respiratory tract diseases]. AB - Asthma is a chronic inflammatory disease of the bronchial tree, characterised by reversible obturation, bronchial mucosal inflammation and bronchial hyperresponsiveness. In the treatment of asthma, most important is avoidance of provoking allergens and regular anti-inflammatory treatment with inhaled glucocorticosteroids. For patients, very important is quick relief from asthma symptoms, this is why they prefer fast and long acting beta 2-agonists as a symptomatic treatment. Optimal control of asthma symptoms can be achieved with combination treatment consisting of inhaled glucocorticosteroid and beta 2 agonist. Budesonide is a corticosteroid that treats the underlying airway inflammation in asthma; formoterol is a fast and long-acting beta 2-agonist that prevents and reverses airway obstruction. Thus budesonide and formoterol have complementary effects, treating two different components of asthma. Clinical studies show that in the treatment of asthma Symbicort is more effective than double dose GCS, due to the budesonide/formoterol synergy. It is at least as effective, well tolerated and safe as its mono-products in corresponding doses given via separate inhalers. More convenient treatment represents an important benefit for patients with asthma. Symbicort is recommended for regular treatment and could also be used "as needed" due to the rapid onset of the bronchodilatory effect of formoterol. PMID- 14524299 TI - [Efficacy and safety of inhaled drugs]. AB - In the paper the foundations are presented of current inhalation therapy, particularly of that applied in childhood. The problems are discussed, concerning deposition of aerosols in the tracheobronchial tree achieved by various methods of aerosol generation. Various factors affecting the bioavailability of the inhaled drugs are defined. Particular attention is paid to the problems of safety of inhaled corticosteroid therapy, stability of the inhaled drugs and their influence on the pulmonary surfactant. PMID- 14524300 TI - [Changes of therapeutic dogma in obstructive lung diseases]. AB - The changes in understanding of the pathogenesis of obstructive diseases of the respiratory tract are responsible for therapeutic strategy. In the paper some discussible current problems is this field have been presented. PMID- 14524301 TI - [Next GINA report. What could be expected?]. AB - The advantages, certain drawbacks and future problems of the next GINA (Global Initiative for Asthma Management and Prevention) elaboration are presented. It is suggested that the next issue is to arrive in 2004 or 2005. PMID- 14524302 TI - [Pharmaco-economics of asthma and chronic obstructive lung disease]. AB - Health economics has become accepted as an important aspect, as it provides useful information on the costs to society associated with particular diseases and the cost-effectiveness of different treatment options. These considerations are particularly important in relation to common and chronic conditions such as asthma and chronic obstructive pulmonary disease. The direct and indirect cost associated with asthma and COPD and trends in the use of outpatient care, drugs and inpatient care were presented in this paper. It should be stressed that asthma therapy has changed from inpatient to ambulatory care, while the treatment of COPD is still based to a higher degree on inpatient care. PMID- 14524303 TI - [Pharmaco-economics of anti-influenza vaccinations]. AB - The types of pharmacoeconomic analysis performed in the assessment of influenza vaccination, including cost-benefit and cost-effectiveness analysis are presented in this study. The studies concerned working adults or high-risk elderly people who were selected from Medline and Embase databases. The primary difference between the two types of vaccination programmes is related to the major type of benefits: direct benefits related to averted hospitalizations for elderly versus indirect benefits of averted production losses for healthy working people. In the group of persons aged over 65 years the disease costs are influenced mainly by complications and hospitalizations. Vaccination is most cost-effective in influenza prophylaxis as compared with other strategies (chemoprophylaxis and treatment using neuraminidase or ion channel inhibitors) and may be recommended from the pharmacoeconomic point of view. PMID- 14524304 TI - [New topical treatment of atopic dermatitis]. AB - Topical inhibitors of calcineurin--tacrolimus and pimecrolimus have recently been approved in the treatment of atopic dermatitis. These are the first topical immunosuppressants other than glucocorticosteroids, awaited in dermatology since approximately 50 years. Both medications present similar mechanism of action, good therapeutic and safety profiles and should be considered as important factors in the treatment of atopic dermatitis. PMID- 14524305 TI - [Recombinant allergens for specific immunotherapy]. AB - Recombinant allergens for allergen specific immunotherapy are accepted in clinical practice. Some allergen variants and epitops have been presented as safety and effective methods influenced the pathomechanism of allergic process. Actual experience with recombinant allergens suggests us a new way in immunotherapy. PMID- 14524306 TI - [Cross-reactivity in immunotherapy]. AB - Side effects of allergen-specific immunotherapy beyond the well-known local and systemic allergic reactions are rare. Cross-reactivity between some groups of inhaled and oral allergens has been documented but not sufficiently explained. Some trials and clinical observations indicate the similarity of allergen subunits and clinical manifestations of this phenomenon. PMID- 14524307 TI - [Practical aspects of specific immunotherapy]. AB - Specific immunotherapy is the only method of causal therapy of allergic diseases. The general principles of this type of intervention are described, including indications, contraindications, efficacy and safety. Attention is paid to the recommended age range of patients, type of hypersensitivity and clinical manifestations that justify undertaking of such intervention. Specific immunotherapy should be supervised by a specialist. PMID- 14524308 TI - [Comparison of injection-related and local immunotherapy]. AB - Allergen-specific immunotherapy is a well-accepted method in the management of respiratory allergic diseases, applied since over 90 years. Traditional immunotherapy is the practice of subcutaneous administering to subjects with allergy, increasing amounts of modified allergen to achieve hyposensitisation. Recently other routes of allergen administration were proposed including bronchial, nasal, oral and sublingual routes. Controlled trials failed to demonstrate the clinical efficacy and the safety of oral and bronchial administration. Nasal immunotherapy is also declining due to limited efficacy and special administration technique. The sublingual route is safe and efficacious. Unfortunately, up to date no well-designed controlled study comparing subcutaneous with sublingual methods of immunotherapy is available. Still several points need to be elucidated, including long lasting efficacy, mechanism of action, optimal dose, adherence to this therapy and cost-effectiveness. However, sublingual immunotherapy is now accepted by WHO as a valid alternative to the subcutaneous route and should be used in all patients who require immunotherapy and do not accept subcutaneous route of allergen administration. One question is presently beyond discussion: it is better to administer sublingual immunotherapy in patients with allergic diseases than not to give immunotherapy at all. PMID- 14524309 TI - [New variants of diagnostic procedures and therapeutic methods in insect venom allergy]. AB - Hymenoptera venom allergy (HVA) is a typical IgE-dependent allergy. Its diagnosis includes confirmation of the presence of specific IgE antibodies to venom allergens (vsIgE), and its treatment is based on developing of immunotolerance (venom immunotherapy, VIT). In most cases of sting anaphylaxis, vsIgE can be found, in some, however, the antibodies are undetectable, in spite of systemic reaction to venom, which may be due to low specificity of diagnostic methods. Diagnostic methods might be improved by the use of recombinant allergens which, being "uncontaminated" by other venom constituents, allow for a precise setup of "individual allergogram." A diagnostic novelty is the tryptase serum level assay that is capable of confirming ex-post anaphylaxis and coincidence of mastocytosis. The diagnostic procedures applied in venom allergy make possible only to prove the existence of allergy while it is impossible to estimate, on their basis, the risk of development and intensification of future reactions. New diagnostic concept that could supplement the imperfect diagnostic armamentarium is still lacking. Clinical practice has proven that VIT is a very effective method of protection of patients with IgE-dependent wasp-sting anaphylaxis, but both the effectiveness and safety of VIT in patients allergic to bee venom are not optimal. The future of VIT is the use of modified, recombinant allergens or their peptide fragments, but clinical data on their effectiveness are unavailable as yet. The safety of VIT can be increased by means of introducing premedication with antihistamines. PMID- 14524310 TI - [Long-term effectiveness of insect venom-specific immunotherapy]. AB - Venom immunotherapy gives high protection from systemic reactions to stings. The question whether VIT provides long term protection after its discontinuation has been raised for many years. The paper presents different factors with an impact on venom immunotherapy outcome. The most important are: age, insect species, severity of the pre-treatment sting reaction, treatment duration, general reactions during VIT to either treatment injections or stings. PMID- 14524311 TI - [Immunotherapy in allergology]. AB - The current state of knowledge is presented of allergenic immunotherapy of atopic allergic diseases emphasizing the fact that desensitisation is an effective and safe method of treatment. Allergenic vaccines, altering the immune profile of patients, correct the deviated immune reactions in atopy. Immunotherapy prevents development of new allergies and the clinical effect is maintained longer than the administered treatment. The technological progress increases the availability of better and ever better standardised vaccines that provide high specificity of administered therapy. PMID- 14524312 TI - [Up to allergy specialists' consideration: did Pharaoh Menes die of a wasp sting?]. AB - Several allergologists interested in allergies to wasp venom, pass a seemingly probable information that the first Egyptian pharaoh, Menes, was allergic to the venom of stinging insects, and died of an insect sting while on a long journey, somewhere in the area of present-day Great Britain. A relief decorating the pharaoh's sarcophagus and the tomb's walls is to prove the historicity of this event. The whole story is a typical hoax. Insufficient historical sources do not confirm the thesis that Menes visited Europe. The tomb which is considered to be his burial site, contained no sarcophagus, reliefs or paintings. The above mentioned relief is a contemporary illustration which should raise suspicions even of a layman who looked at it carefully. It is interesting why so many respectable scientists believe the hoax and quote it without previous verification. PMID- 14524313 TI - [The eye as an effector of allergic reaction]. AB - In this work the symptoms of allergic diseases of the eye were shortly described. The role of mast cells, eosinophils and lymphocytes taking part in allergic reaction was presented. The cellular mechanisms which lead to symptoms of allergic diseases of the eye were discussed. Histopathological changes in the eye tissues and their clinical importance were taken into consideration. The analysis of allergic symptoms of the eye on the basis of the knowledge of pathophysiological processes gives important information both for ophthalmologists and allergologists. PMID- 14524314 TI - [Epidemiology of allergic eye diseases]. AB - Epidemiology of allergic eye diseases has not been sufficiently studied so far. The first statistical studies regarded the coexistence of allergic conjunctivitis together with allergic rhinitis, as rhinoconjunctivitis. Only in the last 10 years eye allergy has been regarded as a separate epidemiological and clinical problem. According to Bonini, seasonal allergic conjunctivitis (SAC) accompanies pollinosis in 95.2%. Buckley's studies revealed symptoms of SAC in 21% of British population and Berdy reported a similar result in 20% of Americans. Weeke estimates that depending on geographical region and age of examined patients, allergic eye diseases occur in 5 to 22% of the population. Among them SAC and perennial allertgic conjunctivitis (PAC) account for up to 50%. A recent Italian study demonstrated an increase of the incidence of allergic eye diseases, which were found in 38% of the studied population, most frequently in young males. Eye allergy presented most frequently as rhinoconjunctivitis (SAC and PAC) (63.7%), and then as atopic keratoconjunctivitis (AKC) (21%) and vernal keratoconjunctivitis (VKC) (15.5%). It seems that the incidence of allergic eye diseases demonstrates a rising tendency, similarly as it has been found in recent years in the case of bronchial asthma, rhinitis or atopic dermatitis. PMID- 14524315 TI - [Surgery in severe forms of allergic eye diseases]. AB - Surgical treatment is performed in allergic eye diseases when drug treatment fails and the disease leads to conjunctival, corneal or palpebral disorders. Surgery procedures can be divided into early (surgery on palpebral conjunctiva, surgery on cornea) and late ones (decreasing tears outflow, plastic surgery on eyelids, cul de sac). PMID- 14524316 TI - [Pterygium: allergic etiology?]. AB - Pterygium is a wedge-shaped fibrovascular growth of conjunctiva that extends onto the cornea. The etiopathogenesis of pterygium is still not fully elucidated. Exposure to ultraviolet light is suggested as one of the most important factors contributing to pterygium development. However, other factors including allergens, harmful chemical substances, wind, dust and pollens also may contribute to its development. The aim of the study was to evaluate the influence of immune reactions on the pathogenesis of pterygium. PMID- 14524317 TI - [Clinical picture, diagnosis and therapy of allergic eye diseases]. AB - Ocular allergic disorders can be a component of systemic or local allergies. The importance of ocular allergy results from its incidence rather than from its severity, however, some of them are vision-threatening. The majority of ocular allergies affect the conjunctiva, eyelids and sometimes cornea that is exposed to the environment and is the place of interaction between allergens and immunocompetent cells. Different types of allergic disorders in the eye may have similar signs and symptoms, but each has its own pathognomonic characteristics, which help to diagnose, differentiate and choose the most suitable therapy. Ocular allergic diseases are classified into six categories: SAC, PAC, VKC, AKC, GPC and ConBC. In 2001 EAACI suggested new classification, also of allergic conjunctivitis, into IgE-mediated and non-IgE-mediated conjunctivitis. IgE mediated conjunctivitis may be divided into intermittent and persistent conjunctivitis. Persistent allergic conjunctivitis is classified into vernal and atopic keratoconjunctivitis. Conjunctivitis contact allergy is a non-IgE form of allergic conjunctivitis. Currently available medications provide safe and effective management of most cases of ocular allergy. Drugs used in the treatment of ocular allergic disorders include mast cell stabilizers, antihistamines, steroids, NSAID's, artificial tears and others. PMID- 14524318 TI - HIV therapeutics, continued: another HIV protease inhibitor approved. PMID- 14524319 TI - Policy watch. Watch the states, too! PMID- 14524320 TI - New medication model helps patients maintain rigorous schedule. PMID- 14524321 TI - New strategies to combat HIV: augmenting antiviral immunity. PMID- 14524323 TI - Editorial comment: why treat three conditions when it is one patient? PMID- 14524322 TI - Triple diagnosis: dual diagnosis and HIV disease, part 2. AB - This review highlights the psychiatric, psychosocial, and substance use treatment issues relevant to the care of patients with triple diagnosis. A discussion of the various psychosocial and psychopharmacologic strategies stresses the importance of integrating medical, psychiatric, and substance use interventions. Components of integrated treatment include a collaborative relationship with various medical services involved in the patient's care, psychotherapeutic interventions tailored to the patient's readiness for change and incorporating motivational strategies, special attention to psychosocial needs, strong emphasis on psychoeducation, and psychopharmacologic management. More research is warranted to evaluate the safety and efficacy of psychopharmacologic and psychotherapeutic modalities in patients with triple diagnosis. PMID- 14524324 TI - Thalidomide-based treatment for HIV-associated multiple myeloma: a case report. AB - The spectrum of hematologic and immunologic abnormalities induced by HIV infection is broad. Although the incidence of HIV-associated B-cell neoplasms has increased, relatively few cases of multiple myeloma have been reported, and even fewer cases have detailed treatment outcome. The case of an HIV-infected man in whom multiple myeloma was diagnosed following progressive anemia and fatigue is described. The patient began treatment consisting of thalidomide, dexamethasone, and clarithromycin, which led to a rapid and dramatic antitumor response. He experienced modest regimen-related toxicities while retaining a normal CD4+ T lymphocyte count and a nondetectable HIV viral load. The immunologic and antitumor effects of thalidomide in the context of multiple myeloma and HIV infection are also briefly reviewed. Given thalidomide's relatively favorable side-effect profile and purported immunologic benefit, further studies of this drug in the treatment of HIV-associated multiple myeloma should be pursued. PMID- 14524325 TI - Editorial comment: multiple myeloma and HIV infection--causal or casual coincidence? PMID- 14524327 TI - Retraction. PMID- 14524326 TI - [Clinical case. Radiologic imaging of chronic sinusitis]. PMID- 14524328 TI - New strategy development tool to improve skilled attendance. PMID- 14524329 TI - Strengthening nursing and midwifery. PMID- 14524330 TI - [Answer to Quiz-H-Q20 (rom J Gastroenterol 2003, vol.12, no.2)]. PMID- 14524331 TI - [Guide to management of non-variceal gastrointestinal hemorrhage]. PMID- 14524332 TI - Oncotic pressure of dietary formulae- an additional characteristic for enteral feeding. PMID- 14524333 TI - Surgery and palliative care. PMID- 14524334 TI - Abstracts of the 19th European Congress of Pathology. Ljubljana, Slovenia, September 6-11, 2003. PMID- 14524335 TI - Abstracts of the 28th Sir Peter Freyer Memorial Lecture and Surgical Symposium in association with the Irish Society of Surgical Oncology. Galway, Ireland, 5-6 September 2003. PMID- 14524336 TI - Abstracts from the 8th Congress of the German Primate Society. October 1-4, 2003, Leipzig, Germany. PMID- 14524338 TI - Abstracts of the British Association for Psychopharmacology. 20-23 July 2003, Cambridge, United Kingdom. PMID- 14524337 TI - The channel in transporters is formed by residues that are rare in transmembrane helices. AB - Transmembrane transport is an essential component of the cell life. Many genes encoding known or putative transport proteins are found in bacterial genomes. In most cases their substrate specificity is not experimentally determined and only approximately predicted by comparative genomic analysis. Even less is known about the 3D structure of transporters. Nevertheless, the published experimental data demonstrate that channel-forming residues determine the substrate specificity of secondary transporters and analysis of these residues would provide better understanding of the transport mechanism. We developed a simple computational method for identification of channel-forming residues in transporter sequences. It is based on the analysis of amino acids frequencies in bacterial secondary transporters. We applied this method to a variety of transmembrane proteins with resolved 3D structure. The predictions are in sufficiently good agreement with the real protein structure. PMID- 14524340 TI - Continuous renal replacement therapies (CRRT). Abstracts of the 8th International Conference on Continuous Renal Replacement Therapies (CRRT). San Diego, California, USA. March 6-8, 2003. PMID- 14524339 TI - Abstracts of the 29th annual meeting of the International Society for Pediatric and Adolescent Diabetes (ISPAD). September 3-6, 2003, Saint-Malo, France. PMID- 14524342 TI - Abstracts of the 21st World Congress on Endourology and SWL 19th Basic Research Symposium. September 21-24, 2003, Montreal, Canada. PMID- 14524343 TI - Abstracts of the 164th Meeting of the Netherlands Anatomical Society. Zeist, The Netherlands. January 4-5, 2002. PMID- 14524341 TI - Lester Snow King. PMID- 14524344 TI - Future perspectives in the treatment of malignant lymphomas: focus on ifosfamide. Proceedings of a symposium. Cap Ferrat, France. October 17, 2002. PMID- 14524345 TI - Abstracts of the 16th Congress of the European College of Neuropsychopharmacology. Prague, Czech Republic, September 20-24, 2003. PMID- 14524346 TI - Prediction of impairment from urine benzoylecgonine concentrations. PMID- 14524348 TI - [Clinical case. Imaging of paranasal sinus trauma]. PMID- 14524347 TI - Tribute to John M. Eisenberg. PMID- 14524349 TI - [Clinical case. Paranasal sinus postoperative imaging]. PMID- 14524350 TI - The foundation and evolution of the Middlesex Hospital's lying-in service, 1745 86. AB - The Middlesex Hospital was founded in 1745, and opened the first British in patient lying-in service in 1747. Men-Midwives were instrumental in founding and supporting the service. The hospital's lying-in service featured prominently in its fundraising literature, and the level of demand from benefactors suggests it was popular. From 1764 the hospital also provided domiciliary services, initially to cope with excess demand and later to compete with domiciliary charities. In 1786 it closed the in-patient services, and from this date provided only domiciliary lying-in services. From 1757, in common with the London lying-in hospitals, the Middlesex Hospital faced competition from a domiciliary charity: The Lying-In Charity for Delivering Poor Married Women in Their Own Homes. Later in the century it also faced competition from dispensaries. This paper describes the foundation and evolution of the Middlesex Hospital's lying-in service, including quantitative information about admissions and about the hospitals income and expenditure during the eighteenth century. It compares the characteristics of domiciliary and in-patient services, to analyse why in-patient services were supported by men-midwives and by benefactors. PMID- 14524351 TI - 'The pregnant women are here for the sake of the teaching institution': The lying in hospital of Gottingen University, 1751 to c. 1830. AB - Drawing on the admission records, the medical casebooks and the publications of its director, this article explores how the University of Gottingen's maternity hospital achieved its three official goals: teaching medical students, training female midwives, and providing shelter for needy parturient women. Since educating medical men was the most important aim of the hospital, the paper particularly focuses on how the demands of instruction shaped day-to-day obstetrical practices, especially under the directorship of Professor Friedrich Benjamin Osiander (1792-1822). He was a keen advocate of the forceps, whereas the first director, Professor Johann Georg Roederer (1751-63), had taken a moderate, that is a much less interventionist, approach to obstetrics. Osiander avowedly was determined to subordinate the parturient women to the demands of the clinic and to treat them as 'living manikins'. In spite of that, there is evidence that the pregnant and parturient women, most of whom were unmarried and from the lower classes, made use of the lying-in hospital for their own purposes, and that sometimes they refused to play the role assigned to them. The link between the maternity hospital and the rise of the man-midwife and of 'scientific' obstetrics appears to have been particularly strong in the case of Gottingen and other German university hospitals, compared with lying-in hospitals in other countries where the link was more indirect. PMID- 14524352 TI - Body weight and self-control in the United States and Britain since the 1950s. AB - Body weight has risen in defiance of health and appearance norms. This is framed in terms of the problem of time-inconsistency and self-control. The social epidemics of overeating and slimming were driven by market forces and the psychology of eating: restrained eating was easily disinhibited by the stresses of new rewards. For men, the rise in body weight was associated with the decline of family eating and exposure to greater food variety. For women, the 'cult of slimming' was associated with mating and workplace competition, driven initially by adverse sex ratios. Food abundance made it difficult for rational consumers to conform with social norms. PMID- 14524354 TI - Molecular cell biology of the endothelium in development and diseases. Abstracts of the 4th Symposium on the Biology of Endothelial Cells. Munich, Germany. July 18-20, 2003. PMID- 14524353 TI - Legal madness in the nineteenth century. AB - Legal sources remain under-exploited in the history of madness, and the legal character of some documents is sometimes unrecognized. This article examines the inter-relations between legal and medical histories of madness, and discusses use and availability of nineteenth-century legal source material relating to criminal insanity, mental incapacity, and the confinement of the insane. PMID- 14524355 TI - Dually eligible for Medicare and Medicaid: two for one or double jeopardy? AB - This issue brief describes the characteristics of the population of individuals known as "dual eligibles," who are eligible for health insurance coverage through both Medicare and Medicaid. It also looks at the differences between "full Medicaid" and "supplemental Medicaid" dual eligibles and the ongoing challenges associated with enrollment and eligibility, integration and coordination, and managed care. The paper presents several examples of integrated care programs designed to better serve the dual-eligible population, including the Program of All-Inclusive Care for the Elderly, Evercare, social health maintenance organizations, and state/federal initiatives such as the Wisconsin Partnership Program, Texas STAR+PLUS, and others. Finally, it considers the implications for dual eligibles of the House and Senate Medicare prescription drug proposals. PMID- 14524357 TI - The safety assessment of novel foods and concepts to determine their safety in use. PMID- 14524356 TI - Refinements to physiologically-based pharmacokinetic model for styrene. PMID- 14524358 TI - Abstracts of the Global Congress of Gynecologic Endoscopy, 32nd annual meeting of the American Association of Gynecologic Laparoscopists. Las Vegas, Nevada, USA. November 19-22, 2003. PMID- 14524359 TI - Abstracts of the 15th Conference of the International Society for Environmental Epidemiology. Perth, Western Australia, 23-26 September 2003. PMID- 14524360 TI - Abstracts of the 4th European Cytogenetics Conference. Bologna, Italy, September 6-9, 2003. PMID- 14524361 TI - Effects of in vitro platelet activation on platelet derived nitric oxide production in healthy humans and in chronic myeloproliferative diseases with elevated platelet counts. AB - Intravascular EDRF-NO production is known to be impaired in some diseases, e.g., diabetes. This phenomenon may also contribute to the development of diabetic vascular disease. More recently the presence of NO synthase (ecNOS, iNOS) have been recognized in human platelets. Platelets produce NO only during activation, even though in minute amounts. This platelet derived NO seems to play an important physiological role, as it inhibits further platelet recruitment quite substantially. In the present report washed platelets isolated from healthy persons and patients with chronic myeloproliferative diseases (CMPD) were exposed to common and physiologically relevant activators (i.e., thrombin, collagen, epinephrine etc.). These tests were carried out in 20 healthy volunteers and 15 patients suffering from myeloproliferative disorders associated with thrombocytosis. As a consequence of pathological platelet function observed in CMPD, the in vitro platelet NO response is impaired in the patient group. One may assume, that reduced platelet NO response, at least in part, may contribute to platelet hyperfunction, angiopathy and thrombotic complications in some cases of CMPD. PMID- 14524362 TI - Association between use of antenatal magnesium sulfate in preterm labor and adverse health outcomes in infants. PMID- 14524363 TI - Impact of midtrimester dilation and evacuation on subsequent pregnancy outcome. PMID- 14524364 TI - Association between the use of antenatal magnesium sulfate in preterm labor and adverse health outcomes in infants. PMID- 14524365 TI - The effect of various insufflation gases on tumor implantation in an animal model. PMID- 14524366 TI - Maternal death at an inner-city hospital, 1949-2000. PMID- 14524367 TI - The Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT). Major outcomes in high-risk hypertensive patients randomized to angiotensin-converting enzyme inhibitor or calcium channel blocker vs diuretic. PMID- 14524368 TI - A history of induced abortion in relation to substance abuse during subsequent pregnancies carried to term. PMID- 14524370 TI - Research on the history of psychiatry. Dissertation abstracts, 2002. PMID- 14524369 TI - Medicare prescription drug legislation: how would it affect beneficiaries? PMID- 14524371 TI - [The influence of changes in the human subjectivity concept of formation of physician-patient relations]. AB - Changes in the human subjectivity concept, expressing influences of cultural and outlook elements on formation of object of the European medicine, have so much affected evolution of the discipline, that it is currently directed to bring an image of a human body to anatomy, to treat a course of a disease in medical aspect and to give to diagnostic processes and therapy a character of means which can be verified in an empirical way. The changes are also reflected in a patient physician relation, which develops in that time. A direct nature of contacts between the treated and the treating persons was taken as grounds of the relation. But reasons of the privileged position of therapists in comparison to patients have changed. The paper aims to show main trends in development of the patient -physician relationship and to represent the changing reasons for superiority of the treating persons who are in a therapeutic relation with a patient. PMID- 14524372 TI - Internal carotid artery dissection after French horn playing. Spontaneous or traumatic event? PMID- 14524373 TI - [The forms of postgraduate education of physicians in Poland in years (1918 1939)]. AB - Postgraduate studies of the Polish physicians between World Wars was not obligatory, except the doctors, who occupied positions in Health Services Administration. They were trained in The State School of Hygiene, in which were performed systematically courses regarding social diseases (tuberculosis, trachoma), organization of health protection. This studies were founded by Department of Health in Ministry of Public Care. The other forms of training were courses of several weeks, regarding general practice or particularly specializations, organized by medical departments of Universities. The aim of this education was completion of the theoretical academic knowledge wish practical skills. The trainings were also organized by medical associations, departments, institutes, hospital wards and other public societies. The postgraduate studies were continued during the domain and foreign medical meetings, by reading medical journals and publications and by practicing in hospital wards. Until the II World War the postgraduate training was not obligatory and the idea of the state institution coordinating this area was not realized. PMID- 14524374 TI - Rational radical neck dissection for oral cancer. PMID- 14524375 TI - [History of teaching of medicine in Gdansk]. AB - The work has aimed at presenting the history of medicine in Gdansk comprising the period from the second half on the 16th century up to the present. The most remote stage of the history was connected with the foundation and development of the Academic Gymnasium of Gdansk (1558) where medicine, general medical sciences and anatomy were taught. Out of those subjects it was the latter that could boast of great achievements owing to famous anatomists lecturing there. It was already in 1613 that the first human dissection was performed in Gdansk, the dissection being also the first in Poland and in Northern Europe. It was here that in 1722 was published the famous anatomical atlas of Jan Adam Kulmus - 'Tabulae anatomicae' - translated for seven languages, including Japanese. Quite a number of graduates of the Academic Gymnasium, having later completed their higher medical education abroad, were coming back to Gdansk and carrying out researches, obtaining excellent results. Among them were: Ludwik von Hammen, Izrael Conradt, Placotomus and others. The Gdansk physicians - Jan Schmiedt (Fabrytius) and Ernest Scheffer were the authors of the first Polish pharmacopoeia (1665). The period from the 18th century up till now has been characterized by great advances in both medical service and hospital management, their crowning achievement being foundation of the Medical Academy in Gdansk, the first, after the war, independent medical university in Poland. PMID- 14524376 TI - [A contribution to the history of MD theses without dissertations. Tauber's case]. AB - The Academic Schools Act 1920 and the ordinance of 1924 pertaining to doctorates provided that one could earn the degree only having submitted a dissertation. The Austrian Act which was in force up to that moment had allowed to receive the degree without writing a thesis. The Jagiellonian University Faculty of Medicine granted doctorates without dissertation up to the end of December 1931. However some doctorates were granted even after that term. PMID- 14524377 TI - China launched an additional SARS research project of National 863 Plan. PMID- 14524378 TI - [Report of the 20th annual reunion of the Andalucia Society of Neurosurgery]. PMID- 14524379 TI - Sampling devices. AB - A number of commonly used samplers are presented in this article. Many samplers have not been discussed because they are used for specific purposes or are considered research tools. Air sampling for microbes may seem like a simple proposal, yet to develop and implement a well-thought out plan that answers questions or hypotheses with a high level of reliability is often a difficult and expensive undertaking. Sampler selection is only one step in this process. The information given in this article, along with the other resources listed, should aid in setting up a useful bioaerosol sampling plan. PMID- 14524380 TI - Floristic zones and aeroallergen diversity. AB - The interplay of geographic, geochemical, and meteorologic factors combines to define distinct floristic zones in North America. Latitude, elevation, Pacific or Atlantic Ocean influence, continental air mass influence, mountains, and hills are contributory geographic factors. Hardiness zones are defined by the nadir of temperature, which strongly affects the survival of individual plant species. There are 12 hardiness zones from the northernmost tundra to the tropics of Mexico. Although it is useful to consider the 10 major floristic zones, the hardiness zones cut across these zones and characterize subregions. A multiplicity of local terrain effects, such as soil porosity and acidity, and sun exposure also impact on plant growth. The ability of plant species, whether woody shrubs and trees, or herbaceous weeds and grasses, to adapt to conditions within the floristic zones determines their range. This article identifies the major aeroallergenic species and the regions in which they are most prevalent. PMID- 14524382 TI - Clinical relevance of spore and pollen counts. AB - Many people with allergies monitor daily pollen and spore counts with the belief that they can act on that information to improve their health. Because many factors can affect personal exposure, the value of community-wide counts for an individual is questionable. These factors include the presence of local pollen and spore sources, diurnal variations, weather effects, air pollution, and a particle-free bioaerosol. To take advantage of bioparticulate counts, the public needs to be informed about their meaning and factors that can influence personal exposure. PMID- 14524381 TI - The urban jungle and allergy. AB - The urban forest is the assemblage of trees, shrubs, and other plants that occupy the urban and suburban zone. In urban areas, the number of potentially allergenic plants has grown rapidly as the diversity of plants increases. The recommended street trees of many cities are allergenic species that are well known to allergy clinicians. Some of the most commonly planted trees in urban zones are known to be the greatest producers of pollen. These trees are situated in close proximity to humans, either at home, at work, or on their travel routes between locations. There are common misconceptions about the plants that do and do not cause allergy. It generally has been considered that insect-pollinated plants with showy flowers are allergy safe; however, when these species are planted in close proximity to people, as they are in urban landscaping, the pollen that leaks from the flowers is often enough to cause an allergic reaction. With increasing emphasis on green space in urban areas, it is advisable to evaluate what is being planted, how much is planted, and the plants' potential for triggering allergy. PMID- 14524383 TI - Meteorologic variables in aerobiology. AB - Although prevalent weather helps define climate, individual weather conditions, such as rain, humidity, wind speed and direction, temperature, or amount of sunshine, may have direct and indirect effects on bioaerosols. Effects may be immediate or cumulative. Precipitation and humidity acutely decrease particle air burden, but sufficient preseason moisture is necessary to assure proper growth of flower buds on perennials and trees and growth of annuals in general. Ambient temperature increase is necessary for anthesis in many plants, and cumulative heat above a threshold value has been linked to onset and intensity of pollination in grasses, weeds, and trees. Wind direction only impacts if there is lack of uniformity in the pollen sources that surround sampling sites. Wind speed may factor in re-entrainment of settled particles or may act to scour the air. Thunderstorms provide a unique sum of factors that greatly may increase aeroallergen burden. Dispersal of mold spores is linked intimately to precipitation and humidity. The effects may be opposed diametrically, however, depending on the type of fungi. Certain ascospores and basidiospores require active rainfall for release of spores, whereas other Deuteromycetes are suppressed by precipitation. PMID- 14524384 TI - Pollen count forecasting. AB - Pollen forecasting is becoming increasingly important to allergists as an adjunct to effective patient care. Forecasts allow patients to avoid exposure to high pollen levels and prompt them to take prophylactic medication and to plan outdoor activities for periods of low pollen levels. Investigators are making progress in developing effective models for daily and seasonal forecasts for important pollen allergens; however, current models are limited to specific geographic areas. Models for the onset of the season for spring tree pollen are based on the chilling and heat units that are required before flowering can occur. Models for pollen season severity are based on regression analysis of preseason meteorologic conditions, and models for daily forecasts are based on the normal pollen curve and responses to day-to-day meteorologic conditions. When winds are favorable, long-distance transport can introduce allergenic pollen types into a local area. The Mountain Cedar Pollen Forecasting model, which combines day-to-day release forecasts at source areas and dispersion forecasts to downwind areas, has been reasonablly successful over the past 4 years. All pollen forecasting models are dependent on accurate meteorologic forecasts, and pollen forecasting will become more accurate as meteorologic forecasts improve. PMID- 14524385 TI - Biology, ecology, and prevalence of dust mites. AB - The house dust mites D. farinae, D. pteronyssinus, and E. maynei are sources of multiple potent allergens in the indoor environment. They are common inhabitants in homes worldwide. Many biologically significant studies have revealed how well adapted these mites are to the microhabitats in homes. Ambient RH is a key factor in determining where these mites are found. Many aspects of the biology of house dust mites are not understood. A greater understanding of the biology of dust mites may reveal new strategies for controlling dust mites and their allergens in homes. PMID- 14524386 TI - Animal danders. AB - Animals release proteins into their surroundings through secretions, as excretions, or as dander. The quantity of dander that is dispersed by cats, dogs, or humans is sufficient to supply food for dust mites and to supply easily measurable quantities of proteins in dust. Fel d 1, Can f 1, and human IgA or IgG can be found in microgram quantities in dust samples. Allergens also can accumulate from the urine of wild or pet rodents. For cats and dogs, the accumulation of dander particles is not related to the cleanliness of the animals. All animals, including humans, provide a fully adequate supply of organic material for bacterial growth in a carpet, provided conditions are sufficiently humid. The authors' preliminary results in Virginia do not find a significant difference in endotoxin between homes with or without animals. The likely explanation for the nonallergic IgG and IgG4 response to cat, dog, or rat allergens is high exposure to proteins from these animals. If the highest levels of cat allergen in a home can result in immunologic tolerance, it is unlikely that primary avoidance would be successful at reducing exposure. The data showing that 80% of Swedish children with cat allergies never had lived with a cat imply that the concentrations of cat allergen in schools or in houses without a cat are sufficient to cause sensitization. Primary prevention would be possible only on a community basis, which is unlikely to occur. Sensitization to cat, rat, dog, or mouse allergens consistently is associated with asthma. In symptomatic children with positive skin test results, there is a strong case for allergen avoidance and a clear need for controlled trials. Controlled trials of avoidance should include houses without cats and schools. Controlling exposure to cat allergens with the cat in situ requires aggressive measures, such as removing reservoirs, washing the cat, and air cleaning. Many allergic or symptomatic children who live with a cat do not have positive skin test results or positive IgE antibodies to cats. Avoidance measures related to animals should be recommended only for individuals with positive skin test results. Increasing evidence shows that exposure to cats, dogs, rats, and other animals can induce a form of immunologic tolerance without causing allergic disease, and it is important to understand why this change occurs with dander allergens rather than with all allergens. The most probable explanations are related to the form and quantity of airborne allergens. PMID- 14524387 TI - Cockroach allergy. AB - The ubiquitous existence of cockroaches and the large-scale domestic infestation seen in inner cities make cockroach proteins a significant indoor allergen and a risk factor for asthma among inner-city residents. Studies have shown that early exposure to high levels of allergen may lead to the development of asthma in individuals with a genetic predisposition to asthma. Although field trials at cockroach abatement do not yield promising results, integrated pest management still remains the best control strategy. In highly susceptible or symptomatic patients, allergen-specific immunotherapy may be beneficial, although data are limited. As molecular techniques improve and recombinant allergens are developed, a more novel form of T-cell-specific immunotherapy may prove to be efficacious without the anaphylactic side effects seen with traditional allergy vaccines. PMID- 14524388 TI - Indoor fungal exposure. AB - Fungi affect humans in complex ways and are capable of eliciting a number of disease responses, such as infectious, allergic, and irritant and toxic effects. Fungal exposure is unequivocally associated with exacerbations of asthma, although the role of fungi in causing the disease is yet to be determined. The association between home dampness and respiratory health effects is strong, and fungal exposure is suspected to be associated with this linkage. Fear of toxin exposures has generated debate over the possible toxic health effects of airborne fungi; however, several recent reviews discount the health impacts of mycotoxin through indoor exposures. Nevertheless, fungal contamination of indoor environments is undesirable. Knowledge of sources and characteristics of fungal spore release and dispersal are important for understanding the processes of exposure. Environmental monitoring for fungi and their disease agents are important aspects of exposure assessment, but few guidelines exist for interpreting their health impacts. Much work is needed in isolating, characterizing and standardizing fungal disease agents to properly assess the prevalence of fungal health effects. PMID- 14524389 TI - Assessment of the indoor environment: evaluation of mold growth indoors. AB - Much attention has been focused on indoor molds; resulting in modest amounts of new research. There is strong evidence of respiratory effects. Although mechanisms are disputed, some of the effect (but not all) is likely to be allergy related. There is some evidence that atopic individuals may be more affected, but many nonatopic individuals also are affected. This area needs more general research and specific research on exposure measures (such as what fungal components should be measured) and on health-effect mechanisms. It is worthwhile to emphasize the practical knowledge that is readily available. Buildings should be designed, built, operated, and occupied so that the buildings stay dry. When this situation does not occur, the environmental and clinical aspects that are observed by competent professionals should both be considered when determining causal relationships. PMID- 14524390 TI - Control of environmental allergens as a therapeutic approach. AB - The first rule of environmental allergen control is that avoidance is appropriate for allergic patients with asthma. Although this rule is obvious to allergists, it frequently is not considered by families and physicians who are not trained in allergy. The next step in environmental avoidance is to determine the specific sensitivity of a patient. Clinically, practitioners have to address reduction of exposure to more than one allergen, and it is likely that the complexity of dealing with several treatment regimes decreases compliance. PMID- 14524391 TI - Cow's milk protein intolerance and childhood constipation. PMID- 14524392 TI - Thyroid nodules and survivors of Hodgkin's disease. PMID- 14524393 TI - Management of labor pain: promoting patient choice. PMID- 14524395 TI - Ivermectin use in scabies. AB - Oral ivermectin is an effective and cost-comparable alternative to topical agents in the treatment of scabies infection. It may be particularly useful in the treatment of severely crusted scabies lesions in immunocompromised patients or when topical therapy has failed. Oral dosing may be more convenient in institutional outbreaks and in the treatment of mentally impaired patients. Ivermectin has been used extensively and safely in the treatment of other parasitic infections, but the U.S. Food and Drug Administration has not approved the drug for the treatment of scabies infection. The safety of oral ivermectin in pregnant and lactating women and young children has yet to be established. PMID- 14524394 TI - Serum tumor markers. AB - Monoclonal antibodies are used to detect serum antigens associated with specific malignancies. These tumor markers are most useful for monitoring response to therapy and detecting early relapse. With the exception of prostate-specific antigen (PSA), tumor markers do not have sufficient sensitivity or specificity for use in screening. Cancer antigen (CA) 27.29 most frequently is used to follow response to therapy in patients with metastatic breast cancer. Carcinoembryonic antigen is used to detect relapse of colorectal cancer, and CA 19-9 may be helpful in establishing the nature of pancreatic masses. CA 125 is useful for evaluating pelvic masses in postmenopausal women, monitoring response to therapy in women with ovarian cancer, and detecting recurrence of this malignancy. Alpha fetoprotein (AFP), a marker for hepatocellular carcinoma, sometimes is used to screen highly selected populations and to assess hepatic masses in patients at particular risk for developing hepatic malignancy. Testing for the beta subunit of human chorionic gonadotropin (beta-hCG) is an integral part of the diagnosis and management of gestational trophoblastic disease. Combined AFP and beta-hCG testing is an essential adjunct in the evaluation and treatment of nonseminomatous germ cell tumors, and in monitoring the response to therapy. AFP and beta-hCG also may be useful in evaluating potential origins of poorly differentiated metastatic cancer. PSA is used to screen for prostate cancer, detect recurrence of the malignancy, and evaluate specific syndromes of adenocarcinoma of unknown primary. PMID- 14524396 TI - Cerebrospinal fluid analysis. AB - Lumbar puncture is frequently performed in primary care. Properly interpreted tests can make cerebrospinal fluid (CSF) a key tool in the diagnosis of a variety of diseases. Proper evaluation of CSF depends on knowing which tests to order, normal ranges for the patient's age, and the test's limitations. Protein level, opening pressure, and CSF-to-serum glucose ratio vary with age. Xanthochromia is most often caused by the presence of blood, but several other conditions should be considered. The presence of blood can be a reliable predictor of subarachnoid hemorrhage but takes several hours to develop. The three-tube method, commonly used to rule out a central nervous system hemorrhage after a "traumatic tap," is not completely reliable. Red blood cells in CSF caused by a traumatic tap or a subarachnoid hemorrhage artificially increase the white blood cell count and protein level, thereby confounding the diagnosis. Diagnostic uncertainty can be decreased by using accepted corrective formulas. White blood cell differential may be misleading early in the course of meningitis, because more than 10 percent of cases with bacterial infection will have an initial lymphocytic predominance and viral meningitis may initially be dominated by neutrophils. Culture is the gold standard for determining the causative organism in meningitis. However, polymerase chain reaction is much faster and more sensitive in some circumstances. Latex agglutination, with high sensitivity but low specificity, may have a role in managing partially treated meningitis. To prove herpetic, cryptococcal, or tubercular infection, special staining techniques or collection methods may be required. PMID- 14524397 TI - The nature and management of labor pain: part I. Nonpharmacologic pain relief. AB - Pain in labor is a nearly universal experience for childbearing women. A recent evidence-based symposium on the nature and management of labor pain brought together family physicians, obstetricians, midwives, obstetric anesthesiologists, and childbirth educators to discuss a series of commissioned systematic reviews. Although management of labor pain plays a relatively minor role in a woman's satisfaction with childbirth compared with the quality of the relationship with her maternity caregiver and the degree of participation she has in decision making, it is an important topic for women and their caregivers. Nonpharmacologic methods of pain relief such as labor support, intradermal water blocks, and warm water baths are effective techniques for management of labor pain. An increased availability of these methods can provide effective alternatives for women in labor. PMID- 14524398 TI - The nature and management of labor pain: part II. Pharmacologic pain relief. AB - A group of family physicians, obstetricians, midwives, obstetric anesthesiologists, and childbirth educators attended an evidence-based symposium in 2001 on the nature and management of labor pain and discussed a series of systematic reviews that focused on methods of labor pain management. Parenteral opioids provide modest pain relief in labor, and little evidence supports the use of one agent over another. Epidural analgesia is used during labor in most large U.S. hospitals, and its use is rapidly increasing in small hospitals. Although epidural analgesia is the most effective form of pain relief, its use is associated with a longer labor, an increased incidence of maternal fever, and increased rates of operative vaginal delivery. The effect of epidural analgesia on rates of cesarean delivery is controversial. Nitrous oxide provides a modest analgesic effect, but it is used less often in the United States than in other developed nations. Paracervical block provides effective analgesia in the first stage of labor, but its use is limited by postblock bradycardia. Research is needed regarding which pain-relief options women would choose if they were offered a range of choices beyond epidural analgesia or parenteral opioids. PMID- 14524399 TI - Information from your family doctor. Labor pain: what to expect and ways to relieve pain. PMID- 14524400 TI - Differential diagnosis and treatment of hearing loss. AB - Hearing loss is a common problem that can occur at any age and makes verbal communication difficult. The ear is divided anatomically into three sections (external, middle, and inner), and pathology contributing to hearing loss may strike one or more sections. Hearing loss can be categorized as conductive, sensorineural, or both. Leading causes of conductive hearing loss include cerumen impaction, otitis media, and otosclerosis. Leading causes of sensorineural hearing loss include inherited disorders, noise exposure, and presbycusis. An understanding of the indications for medical management, surgical treatment, and amplification can help the family physician provide more effective care for these patients. PMID- 14524402 TI - Information from your family doctor. What can I do for dry, itchy skin? PMID- 14524401 TI - Pruritus. AB - Pruritus is a common manifestation of dermatologic diseases, including xerotic eczema, atopic dermatitis, and allergic contact dermatitis. Effective treatment of pruritus can prevent scratch-induced complications such as lichen simplex chronicus and impetigo. Patients, particularly elderly adults, with severe pruritus that does not respond to conservative therapy should be evaluated for an underlying systemic disease. Causes of systemic pruritus include uremia, cholestasis, polycythemia vera, Hodgkin's lymphoma, hyperthyroidism, and human immunodeficiency virus (HIV) infection. Skin scraping, biopsy, or culture may be indicated if skin lesions are present. Diagnostic testing is directed by the clinical evaluation and may include a complete blood count and measurement of thyroid-stimulating hormone, serum bilirubin, alkaline phosphatase, serum creatinine, and blood urea nitrogen levels. Chest radiography and testing for HIV infection may be indicated in some patients. Management of nonspecific pruritus is directed mostly at preventing xerosis. Management of disease-specific pruritus has been established for certain systemic conditions, including uremia and cholestasis. PMID- 14524403 TI - Diagnostic approach to polyarticular joint pain. AB - Identifying the cause of polyarticular joint pain can be difficult because of the extensive differential diagnosis. A thorough history and a complete physical examination are essential. Six clinical factors are helpful in narrowing the possible causes: disease chronology, inflammation, distribution, extra-articular manifestations, disease course, and patient demographics. Patients with an inflammatory arthritis are more likely to have palpable synovitis and morning stiffness; if the condition is severe, they may have fever, weight loss, and fatigue. Viral infections, crystal-induced arthritis, and serum sickness reactions are common causes of acute, self-limited polyarthritis. Because chronic arthritides may present abruptly, they need to be considered in patients who present with acute polyarticular joint pain. Joint palpation can help to distinguish inflammatory synovitis from the bony hypertrophy and crepitus that typically occur with osteoarthritis. Extra-articular manifestations of rheumatologic disease may be helpful in arriving at a more specific diagnosis. Many classic rheumatologic laboratory tests are nonspecific. A complete blood count, urinalysis, and a metabolic panel may provide more useful diagnostic clues. Plain-film radiographs may demonstrate classic findings of specific rheumatologic diseases; however, radiographs can be normal or only show nonspecific changes early in the disease process. PMID- 14524404 TI - Information from your family doctor. Pregnancy: prenatal care. PMID- 14524405 TI - Information from your family doctor. Pregnancy and exercise. PMID- 14524406 TI - Information from your family doctor. Respiratory infections during pregnancy. PMID- 14524407 TI - Evaluation of epigastric discomfort and management of dyspepsia and GERD. PMID- 14524408 TI - Documentation of animal quality. PMID- 14524409 TI - Aspects of common marmoset basic biology and life history important for biomedical research. AB - While common marmosets are increasingly used as alternative primate models in biomedical research, their life history, specialized behavior and unique physiology are not well known. This paper describes important marmoset attributes that are particularly relevant for biomedical research, including reproduction, neurobiology, immunology, endocrine signaling, obesity and aging, in addition to fetal and postnatal development. While common marmosets exhibit characteristic anthropoid primate traits, they clearly differ from Old World primates and humans in a variety of functions, including reproduction, endocrine signaling and immunology. These differences, however, permit the use of common marmosets in unconventional research strategies targeted on human pathology. PMID- 14524410 TI - Husbandry, handling, and nutrition for marmosets. AB - Marmosets, especially Callithrix jacchus, have become an established part of the laboratory animal community. Information on marmoset life history, behavior, and diet acquired from experience with natural and captive habitats has increased, but the early information from workers with colonies, principally those of tamarins, has led to some common perceptions about how to house, handle, and especially, feed callitrichids that may not apply to marmoset requirements. The availability of commercially produced, almost-complete base diet components and a wider variety of cage construction materials, combined with the recent emphasis on the integration of engineering and performance standards for housing, have made captive life and the implementation of research requirements better for the animals and the people that work with them. We will review some of the routine aspects of husbandry, handling, and nutrition for marmoset monkeys maintained in a research setting. PMID- 14524411 TI - Sample collection and restraint techniques used for common marmosets (Callithrix jacchus). AB - The common marmoset (Callithrix jacchus) is a small-bodied, adaptable New World primate from secondary forests in Brazil that is used in various types of research, such as reproductive biology, neuroendocrinology, behavioral research, neuroscience, infectious disease, and drug development. Because of their small body size, adaptability to a variety of conditions, unique physiologic characteristics, family social structure, and calm demeanor, they have become the primate of choice for certain research areas. However, because of their small body size, the amount and type of samples that can be obtained from them can prove to be challenging. The objective of the study reported here is to review some techniques developed for obtaining samples or data from marmoset monkeys in a variety of research settings and includes restraint methods that work well for the type of sample collection required. PMID- 14524412 TI - Reproduction in captive common marmosets (Callithrix jacchus). AB - Though sexual maturation may begin at around one year of age, first successful reproduction of the common marmoset (Callithrix jacchus) is likely to be later, and it is generally recommended that animals not be mated before 1.5 years of age. The average gestation period is estimated to be 143 to 144 days. A crown rump length measurement taken by use of ultrasonography during the linear, rapid, prenatal growth phase (between approx. days 60 and 95) can be compared against standard growth curves to estimate delivery date to within 3 to 4 days, on average. Marmosets produce more young per delivery than does any other anthropoid primate, and have more variation in litter size. Many long-established colonies report that triplets are the most common litter size, and there is documented association between higher maternal body weight and higher ovulation numbers. Higher litter sizes generally do not generate higher numbers of viable young. Marmosets are unusual among primates in having a postpartum ovulation that typically results in conception and successful delivery; reported median inter birth intervals range from 154 to 162 days. However, pregnancy losses are quite common; one study of a large breeding colony indicated 50 percent loss between conception and term delivery. The average life span for breeding females is around six years; the range of reported average lifetime number of litters for a breeding pair is 3.45 to 4.0. Our purpose is to provide an overview of reproduction in the common marmoset, including basic reproductive life history, lactation and weaning, social housing requirements, and common problems encountered in the captive breeding of this species. A brief comparison between marmoset and tamarin reproduction also will be provided. PMID- 14524413 TI - Clinical care and diseases of the common marmoset (Callithrix jacchus). AB - The common marmoset (Callithrix jacchus) is a small New World primate that is native to eastern Brazil and has been used in biomedical research since the early 1960s. Use of this species for research purposes continues to grow at a rapid pace as they are a viable alternative to other nonhuman primate species. We discuss clinical care, such as preventative medicine, anesthesia, and routine clinical procedures. Important viral, bacterial, parasitic, and gastrointestinal tract diseases are discussed, with relevant details on cause, transmission, pathologic changes, diagnosis, and treatment. Current problems affecting marmoset health and research in the modern vivarium are emphasized. PMID- 14524414 TI - Marmoset models commonly used in biomedical research. AB - The common marmoset (Callithrix jacchus) is a small, nonendangered New World primate that is native to Brazil and has been used extensively in biomedical research. Historically the common marmoset has been used in neuroscience, reproductive biology, infectious disease, and behavioral research. Recently, the species has been used increasingly in drug development and safety assessment. Advantages relate to size, cost, husbandry, and biosafety issues as well as unique physiologic differences that may be used in model development. Availability and ease of breeding in captivity suggest that they may represent an alternative species to more traditional nonhuman primates. The marmoset models commonly used in biomedical research are presented, with emphasis on those that may provide an alternative to traditional nonhuman primate species. PMID- 14524415 TI - Study of low-temperature (4 degrees C) transport of mouse two-cell embryos enclosed in oviducts. AB - PURPOSE: We examined usefulness of a mouse embryo transportation system for low temperature transport of oviducts containing mouse two-cell embryos. METHODS: Oviducts containing two-cell mouse embryos were stored at 4 degrees C for 36 h. After that, embryos were collected and cultured for 96 h in Potassium Simplex Optimized Medium (KSOM) medium and evaluated for their rate of development to hatched blastocysts. Embryos were transferred to recipients, and the rate of survival to live young was investigated. The oviducts were then transported from Yamagata to Kumamoto (distance of approx. 1,000 km). At the destination, embryos were implanted in recipient dams and were studied to evaluate their survival to live young. RESULTS: After preservation for 36 h at 4 degrees C, 68.3% of two cell embryos developed to hatched blastocysts. As a result of transplanting 546 embryos into 25 recipients, 109 normal live young mice were obtained; the rate of development was 20.0%. Results of oviduct transport from Yamagata to Kumamoto indicated that 30.2% of transplanted embryos developed to live young. CONCLUSION: Low-temperature transport of two-cell embryos in oviducts is useful as a method of shipping mouse embryos between institutes. PMID- 14524416 TI - Susceptibility of irradiated B6D2F1/J mice to Klebsiella pneumoniae administered intratracheally: a pulmonary infection model in an immunocompromised host. AB - Bacteria such as Klebsiella pneumoniae can invade and colonize an immunocompromised host and complicate clinical recovery. In the study reported here, an experimental model of induced pneumonia was developed in 60Co gamma photon-irradiated mice for the purpose of evaluating efficacy of therapeutic agents. The model was characterized by use of probit analysis of bacterial dose, and microbiologic, and histopathologic results. Bacterial colony-forming-unit (CFU) values producing 50% mortality within 30 days (LD50/30) and their 95% confidence intervals were 4.0 x 10(4) [1.7 x 10(4) - 8.9 x 10(4)] for 0-Gray (Gy) irradiated mice, 1.9 x 10(4) [7.0 x 10(3) - 4.8 x 10(4)] for 5-Gy-irradiated mice, and 1.0 x 10(3) [2.8 x 10(2) - 3.3 x 10(3)] for 7-Gy-irradiated mice. Probit regression line fits calculated by use of an iterative, weighted least squares fit, were used to assess a dose-modifying factor (DMF). The DMFs for mortality, compared with that for the 0-Gy dose, with their 95% confidence intervals, were 2.2 [0.63 - 7.7] for the 5-Gy and 38.9 [9.6 -165.0] for 7-Gy doses. The 5-Gy probit line did not significantly differ (P = 0.21) from the 0-Gy probit line (dose ratios did not significantly differ from 1), whereas the 7-Gy probit line differed significantly from the 0-Gy probit line (P < 0.001). These results demonstrate that 7-Gy 60Co gamma-photon radiation in combination with intratracheal K. pneumoniae challenge induces a valid pulmonary infection model in immunocompromised female B6D2F1/J mice. PMID- 14524417 TI - Qualitative and quantitative differences in normal vaginal flora of conventionally reared mice, rats, hamsters, rabbits, and dogs. AB - We examined quantitatively the vaginal flora of conventionally reared mice, rats, hamsters, rabbits and dogs, species that are widely used as laboratory animals. Vaginal specimens were examined according to the method of analyzing intestinal flora (Mitsuoka's procedure). The total number of bacteria (aerobes and anaerobes) and the prevalence of specific bacteria were determined. The total number of bacteria was highest during estrus and lowest during diestrus or anestrus in mice, rats, hamsters, and dogs. The most predominant bacteria during estrus were streptococci in mice; gram-negative rods (GNR), streptococci, and members of the family Bacteroidaceae in rats; GNR, Bacteroidaceae and gram positive anaerobic cocci in hamsters, and Bacteroidaceae in dogs. The increase in the total number of bacteria during estrus was caused by an increase of predominant bacteria in the vagina. Aerobes were more predominant than anaerobes in mice, and number of aerobes was comparable to that of anaerobes in rats and dogs. On the other hand, in hamsters, anaerobes were more predominant than aerobes and the total number of bacteria was highest among the laboratory animals (mice, rats, hamsters, rabbits, and dogs). However, in rabbits, bacteria were not isolated from about 90% of the vaginal specimens. Rabbits do not have cyclic reproductive stages and are usually in precoital status in the laboratory. In precoital rabbits, vaginal epithelium manifests few signs of secretion. Therefore, we suspect that the vaginal environment in precoital rabbits is comparable to that during diestrus or anestrus in mice, rats, hamsters, and dogs. These results suggest that the vaginal flora of laboratory animals is influenced by the estrous cycle, and probably by mucous secretion. Our data imply that vaginal flora differ among laboratory animals species, and researchers need to take into consideration the estrous cycle of laboratory animals when studying their vaginal flora. PMID- 14524418 TI - Normal hematologic and serum clinical chemistry values for captive chimpanzees (Pan troglodytes). AB - In the study reported here, reference intervals for hematologic and serum clinical chemistry variables in the chimpanzee (Pan troglodytes) were developed and characterized. Data were collected longitudinally across a 10-year period for 86 subjects at the Primate Foundation of Arizona (PFA). Variables included nine standard hematologic and 25 standard serum clinical chemistry values. An analysis of variance (ANOVA) was used to test for main effects by age and sex. In addition, PFA mean and range values were compared with those published for humans and six other chimpanzee colonies. The ANOVA results suggest an age effect on hematologic (mean corpuscular hemoglobin, mean corpuscular volume, neutrophils) and serum clinical chemical (creatinine, total protein, globulin, tryglycerides, direct bilirubin, iron, (gamma-glutamyltransferase, alanine transaminase, creatine kinase) values. In addition, sex had a main effect on several variables (red blood cells, hemoglobin concentration, hematocrit, uric acid and sodium concentrations, and aspartate transminase and creatine kinase activities); values for males were greater than those for females. Further, human and chimpanzee mean and range values often were indistinguishable from one another. However, changes in human and chimpanzee values associated with age differ and suggest that hematologic and serum clinical chemistry values may be differentially affected by physical and sexual maturation in humans and chimpanzees. PMID- 14524419 TI - Spontaneous pituitary abnormalities and mammary hyperplasia in FVB/NCr mice: implications for mouse modeling. AB - The FVB/N mouse strain is widely used in the generation of transgenic mouse models. We have observed that mammary glands of wild-type virgin female FVB/NCr mice frequently have the morphologic and histologic appearance of a gland during pregnancy. By 13 months of age, the mammary glands of more than 40% of the mice examined had lobuloalveolar hyperplasia that was characterized by the presence of secretory alveoli and distended ducts apparently containing secretory material. The prevalence of this phenotype further increased with age. The mammary phenotype was highly correlated with the presence of proliferative, prolactin secreting lesions in the pituitary gland. In mice aged 18 to 23 months, hyperplasia of the pars distalis was seen in 11 of 21 mice (52%), and a further 4 of 21 mice (19%) had pituitary adenomas. Pituitary hyperplasia was already evident in some mice as young as nine months. The pituitary phenotype was also associated with high prevalence (4/6 mice) of spontaneous mammary tumors in aged multiparous, but not virgin FVB/NCr mice. This high prevalence of pituitary abnormalities and their effects on the mammary gland have important consequences for the interpretation of new phenotypes generated in transgenic models using this mouse substrain. PMID- 14524421 TI - Confirmed persistent mouse hepatitis virus infection and transmission by mice with a targeted null mutation of tumor necrosis factor to sentinel mice, using short-term exposure. AB - Mouse hepatitis virus (MHV) infection in immunocompetent mice is typically self limiting, and transmission is short lived. With the recent surge in the development of genetically engineered mutant mice with alterations in immune system components, however, MHV clearance may be disrupted. We report confirmed persistent transmission of MHV from tumor necrosis factor (TNF) knockout mice, B6.129S1-Tnftm1Lj (TNF -/-), to nude and immunocompetent sentinel mice over a period of five months. Infection with MHV was confirmed in nude sentinel mice by use of reverse transcriptase-polymerase chain reaction (RT-PCR) detection of viral RNA in ascending colon and feces. The RT-PCR-analyzed specimens recovered from sentinel animals were sequenced, and 92% homology to the N region of the MHV strain S genome was documented. In addition, immunocompetent mice had evidence of seroconversion to MHV infection and RT-PCR-positive fecal and ascending colon specimens after only 24 h of direct contact with the TNF -/- mice. To the authors' knowledge, this is the first reported experimental evidence that MHV transmission can occur for several months, from persistently infected mice to sentinel mice, over a short-term exposure period. PMID- 14524420 TI - Persistent mammary hyperplasia in FVB/N mice. AB - The inbred FVB/N mouse strain is widely used for creating transgenic mice. Over the past decade, persistent mammary hyperplasia has been detected in many multiparous FVB/N female mice sent to the University of California, Davis (UCD) Mutant Mouse Pathology Laboratory (MMPL) by a number of different laboratories. However, the experimental details concerning most specimens were not always available. To confirm these empiric findings, experiments were carried out to evaluate the mammary glands of FVB/N mice under controlled conditions. Persistent mammary hyperplasia that related to parity was found. Weeks after their first to fourth pregnancy, 10 FVB/N female mice from the Lawrence Berkeley National Laboratory (LBNL) colony were studied and the mammary glands were evaluated. The percentage of fat pad filled was estimated, using image analysis. Serum samples and the pituitary gland from other FVB/N mice from the LBNL were assayed for prolactin concentration. Multiparous FVB/N females consistently had persistent mammary hyperplasia. Four of seven females in the LBNL colony had hyperplasia after three pregnancies. A few foci of squamous nodules and sporadic carcinomas also were observed. Thus, some FVB/N females may have persistent mammary hyperplasia after three pregnancies without detectable pituitary abnormalities. Mammary carcinomas also may develop sporadically. These background phenotypes must be considered when interpreting the effect of genetic manipulation in FVB/N mice. PMID- 14524422 TI - Source of a micro-nutrient in a semi-synthetic basal diet as a causative factor in inducing urinary calculi in rats and its inhibition by PSC 833, a potent inhibitor of P-glycoprotein. AB - We report a serendipitous finding of urinary calculi in rats fed a semi-synthetic basal diet. This observation was made during ongoing studies to evaluate the inhibitory effect of PSC 833, a potent inhibitor of P-glycoprotein, on development of tumors in rodent tumor model systems. A large number of specific pathogen-free (SPF) female Sprague-Dawley and SPF male Fischer 344 rats being fed the diet were euthanized when it became evident clinically that they were uremic. At necropsy, the renal pelvis, ureters, and urinary bladder contained numerous calculi. The presence of urinary calculi was determined to be related to the source of a Food Chemical Codex grade of choline bitartrate. Rats being fed the same basal diet containing the United States Pharmacopia grade of choline bitartrate failed to develop urinary calculi. Interestingly, rats treated with the P-glycoprotein inhibitor were at significantly reduced risk of developing urinary calculi. This finding highlights how something seemingly innocuous as a minor dietary constituent can have a profound impact and, thereby, affect experimental outcome. PMID- 14524423 TI - Gatekeeping in child welfare: a comparative study of intake decisionmaking by social workers in Canada and Sweden. AB - This article details findings from social workers in Sweden and Canada, illuminating similarities and differences in gatekeeping in child welfare and child protection. Analysis revealed different patterns of inclusion and exclusion. Swedish child welfare includes a greater readiness to intervene with more resources and measures. Gatekeeping is assessment driven and focused on family preservation. In Canada, only the most needy children are eligible for a limited range of services. Gatekeeping is structure driven and narrowly focused on protection. Analyses of evidence-based research to improve outcomes for children and families must include comparisons of how different structural orientations influence management of referrals at intake. The authors discuss the implications of these findings. PMID- 14524425 TI - Characteristics of evidence-based child maltreatment interventions. AB - This project summarizes, using a treatment protocol review technique, characteristics of effective interventions from nine studies of child maltreatment that examined recovery from abuse or the effects of maltreatment on child and parent outcomes. Results suggest that stronger effects are yielded by targeting parents and the parent-child interaction context in home-based settings during early childhood, designing multicomponent interventions delivered by professionals for teaching parenting competency skills, and targeting families of higher risk children. PMID- 14524424 TI - The empirical basis of risk assessment in child welfare: the accuracy of risk assessment and clinical judgment. AB - The importance of risk assessment is juxtaposed with the lack of empirical support regarding the validity of risk inventories. This study compared risk ratings of one risk assessment tool to decisions made by case managers. The researchers sampled 450 children and compared predictive utility of risk assessment to child protection decisions. Risk assessment was consistent with clinical judgment in 74% to 81% of cases, more than previously reported in studies of risk assessment validity. Further analyses identified discriminate functions at the instrument's category and individual-item levels. The results have implications for the validity of the instrument and its utility in child welfare. PMID- 14524426 TI - Parental views of in-home services: what predicts satisfaction with child welfare workers? AB - Evidence on client satisfaction deserves consideration in the design of child welfare policies, programs, and practices. Data in this study come from the National Survey of Child and Adolescent Well-Being. Clients receiving in-home services reported moderate levels of satisfaction with their child welfare workers. Caregiver reports of having less than two child welfare workers, having more recent contact, and receiving timely, responsive services were associated with higher perceived quality of relationships with child welfare workers. The child welfare workers' reports of cooperativeness by the caregiver were also associated with higher caregiver-reported relationship quality. PMID- 14524428 TI - Building evidence to promote educational competence of youth in foster care. AB - Although the academic difficulties of students living in foster care are well documented, few studies have examined factors influencing academic achievement of youth in foster care. This article reports the results of a study of educational competence in a sample of 152 foster youth in one midwestern state. Using five standardized instruments, the study explored predictors of reading ability of youth in foster care. Multiple regression analyses indicated that four factors- aspiration for higher education, placement in kinship care, participation in extracurricular activities, and drug use--accounted for 39% of the variance in reading ability scores. The article discusses implications of these findings for practice, research, and policy development. PMID- 14524427 TI - Evidence-based practice in community-based child welfare systems. AB - The immaturity of the evidence base in some areas of child welfare practice demands that practitioners approach the development and refinement of practice in a systematic way that explicitly seeks to build on and learn from experience. The efforts of public child welfare agencies in Boston and Cleveland show that it is possible to draw on a broader base of knowledge and experience while responding to unique conditions and circumstances in each agency's community. PMID- 14524429 TI - School-based early intervention and child well-being in the Chicago Longitudinal Study. AB - Chicago Child-Parent Centers provide comprehensive education and family support services to young, low-income children. Using data from 1,539 children in the Chicago Longitudinal Study, preschool participation was associated with significantly higher levels of school readiness, achievement, and educational attainment, and with lower rates of child maltreatment, juvenile delinquency, special education placement, and grade retention. Every dollar invested in the preschool program returned 7.14 dollars to society at large. School-age intervention also provided economic returns that exceeded costs. Findings provided strong evidence that large-scale, public, early interventions can enhance children's well-being. The authors recommend greater investments in programs with elements similar to the Child-Parent Centers. PMID- 14524430 TI - Fate of estrogens in a municipal sewage treatment plant. AB - The fate of the highly potent endocrine disrupters estrone (E1), 17beta-estradiol (E2), and 17alpha-ethinylestradiol (EE2) was investigated in mechanical and biological sewage treatment as well as in sewage-sludge treatment at a municipal German sewage treatment plant (STP). The main outcome of the study was that a common municipal STP with an activated sludge system for nitrification and denitrification including sludge recirculation can appreciably eliminate natural and synthetic estrogens. As a consequence, the endocrine effects of biota in the receiving waters should be significantly reduced. All estrogen concentrations decreased gradually along the treatment train. In the STP effluent, the steroid estrogen concentrations were always below the quantification limit of 1 ng/L. The elimination efficiency of the natural estrogens (E1 and E2) exceeded 98%, and EE2 was reduced by more than 90%. The natural estrogens were largely degraded biologically in the denitrifying and aerated nitrifying tanks of the activated sludge system, whereas EE2 was only degraded in the nitrifying tank. Only about 5% of the estrogens are sorbed onto digested sewage sludge. It is very likely that conjugates (glucuronides and sulfates) of the estrogens were cleaved into the parent compounds mainly in the first denitrification tank. PMID- 14524431 TI - Evidence for the absence of Staphylococcus aureus in land applied biosolids. AB - Staphylococcus aureus is an important human pathogen both within the hospital setting and as a community-acquired infection. Recently there has been concern that land applied biosolids may transmit S. aureus. However, no scientific data are available to document whether biosolids are a source of S. aureus. To determine if S. aureus is present in biosolids, we collected samples from 15 sites across the United States. Samples analyzed were as follows: 3 raw untreated sewage samples and 2 undigested primary sewage sludge samples; 23 different biosolid samples; and 27 aerosols obtained during biosolid land application (biosolid aerosols). Although S. aureus were detected in raw sewage samples, none were found in any of the treated biosolids nor in any biosolid aerosol samples. These results suggest that biosolids are not a likely source of S. aureus human exposure or infection. PMID- 14524432 TI - Nonylphenol ethoxylates and other additives in aircraft deicers, antiicers, and waters receiving airport runoff. AB - Samples of nine different formulations of aircraft deicer and antiicer fluids (ADAF) were screened for the presence of selected surfactants. Nonylphenol ethoxylates (NPnEO) were identified in three ADAF formulations, octylphenol ethoxylates were identified in two formulations, and six formulations contained alcohol ethoxylates. A preliminary field study was conducted at General Mitchell International Airport, Milwaukee, WI, to quantify NPnEO (n = 1-15) and one of its byproducts, nonylphenol (NP), in airport runoff. Samples were collected from two airport outfalls, from the receiving stream, and from an upstream reference site during intensive ADAF application events. NPnEO was measured at concentrations up to 1190microg/L in airport outfall samples, up to 77 ug/L in samples from the receiving stream and less than 5.0 microg/L from the upstream reference. Concentrations of glycol and other ADAF-related constituents, including NPnEO, were reduced by approximately 1 order of magnitude between the outfall sites and the receiving stream site; however, concentrations of NP in the receiving stream remained similar to those from the outfalls (< 0.04 microg/L at the upstream reference, 0.98 and 7.67 microg/L at outfalls, and 3.89 microg/L in the receiving stream). The field data suggest that NP is generated through degradation of NPnEO from airport runoff. PMID- 14524433 TI - Variable air temperature response of gas-phase atmospheric polychlorinated biphenyls near a former manufacturing facility. AB - Many investigations of gas-phase atmospheric PCB show a strong relationship between concentration and air temperature, especially near PCB sources. Comparative gas-phase atmospheric PCB trends during an annual temperature regime at two sites near a former PCB manufacturing plant and nearby PCB landfills in Anniston, AL, indicate a departure from this trend. The Mars Hill sampling site, located closest to the plant and landfills, shows an annual average sigmaPCB concentration of 27 ng m(-3) (ranging from 8.7 to 82 ng m(-3)) three times the average at Carter, 1.5 km away (9 ng m(-3), ranging from 1.1 to 39). However, total PCB and congener concentrations vary more with air temperature at Carter where PCB are evaporating from surfaces during warmer weather. The slopes of the Clausius-Clapeyron plots of 18 of the most concentrated congeners representing dichloro- through heptachlorobiphenyl homologues are significantly higher at the Carter site. While some of the atmospheric PCB at Mars Hill is derived from ground surface evaporation, the source of much of it apparently is the material buried in the landfills, which has different thermal properties than surface materials and is not in equilibrium with air temperature. PMID- 14524434 TI - Biomarkers in fish from Prince William Sound and the Gulf of Alaska: 1999-2000. AB - To test the hypothesis that biomarker levels in fish collected at Prince William Sound (PWS) sites impacted by the 1989 Exxon Valdez oil spill were higher than those collected at unimpacted sites, a 1999-2000 study collected five fish species and associated benthic sediments from 21 sites in PWS and the eastern Gulf of Alaska (GOA). PWS sites were divided in three oiling categories based upon 1989 shoreline assessments: nonspill path (NSP), spill path oiled (SPO), and spill path not oiled (SPNO). Rockfish (N = 177), rock sole (N = 30), and kelp greenling (N = 49) were collected at near-shore locations (approximately 50-500 m from shore); Pacific halibut (N = 131) and Pacific cod (N = 81) were collected further offshore (approximately 500-7000 m). Fish were assayed for bile fluorescent aromatic contaminants (FAC) and cytochrome P4501A (CYP1A) levels measured as liver ethoxyresorufin O-deethylase (EROD) activity and by immunohistochemistry (IHC) of various tissues. For all species studied at all sites, bile FAC concentrations and CYP1A levels were low and in the same range for fish collected at PWS SPO and SPNO sites relative to NSP sites in PWS and the GOA. Consequently, the hypothesis is rejected for the species studied. The bile FAC results further indicate a pervasive exposure of fish at all sites, including those in the GOA far removed from the effects of the spill, to low levels of polycyclic aromatic hydrocarbons. Analysis of the benthic sediments indicates that the probable sources of this exposure are petrogenic hydrocarbons derived from natural oil seeps and eroding sedimentary rocks in the eastern GOA. PMID- 14524435 TI - Delineation of estuarine management areas using multivariate geostatistics: the case of Sado Estuary. AB - The Sado Estuary is a coastal zone located in the south of Portugal where conflicts between conservation and development exist because of its location near industrialized urban zones and its designation as a natural reserve. The aim of this paper is to evaluate a set of multivariate geostatistical approaches to delineate spatially contiguous regions of sediment structure for Sado Estuary. These areas will be the supporting infrastructure of an environmental management system for this estuary. The boundaries of each homogeneous area were derived from three sediment characterization attributes through three different approaches: (1) cluster analysis of dissimilarity matrix function of geographical separation followed by indicator kriging of the cluster data, (2) discriminant analysis of kriged values of the three sediment attributes, and (3) a combination of methods 1 and 2. Final maximum likelihood classification was integrated into a geographical information system. All methods generated fairly spatially contiguous management areas that reproduce well the environment of the estuary. Map comparison techniques based on kappa statistics showed thatthe resultant three maps are similar, supporting the choice of any of the methods as appropriate for management of the Sado Estuary. However, the results of method 1 seem to be in better agreement with estuary behavior, assessment of contamination sources, and previous work conducted at this site. PMID- 14524436 TI - Airborne brake wear debris: size distributions, composition, and a comparison of dynamometer and vehicle tests. AB - Particle size distributions of light-duty vehicle brake wear debris are reported with careful attention paid to avoid sampling biases. Electrical low-pressure impactor and micro-orifice uniform deposit impactor measurements yield consistent size distributions, and the net particulate matter mass from each method is in good agreement with gravimetric filter measurements. The mass mean diameter of wear debris from braking events representative of urban driving is 6 microm, and the number-weighted mean is 1-2 microm for three currently used classes of lining materials: low metallic, semimetallic, and non-asbestos organic (NAO). In contrast, the wear rates are very material dependent, both in number and mass of particles, with 3-4 times higher emissions observed from the low metallic linings as compared to the semimetallic and NAO linings. Wind tunnel and test track measurements demonstrate the appearance of micron size particles that correlate with braking events, with approximately 50% of the wear debris being airborne for the test vehicle in this study. Elemental analysis of the wear debris reveals a consistent presence of the elements Fe, Cu, and Ba in both dynamometer and test track samples. PMID- 14524437 TI - On-road measurement of particle emission in the exhaust plume of a diesel passenger car. AB - Particle size distributions were measured under real world dilution conditions in the exhaust plume of a diesel passenger car closely followed by a mobile laboratory on a high speed test track. Under carefully controlled conditions the exhaust plume was continuously sampled and analyzed inside the mobile laboratory. Exhaust particle size distribution data were recorded together with exhaust gas concentrations, i.e., CO, CO2, and NO(x), and compared to data obtained from the same vehicle tested on a chassis dynamometer. Good agreement was found for the soot mode particles which occurred at a geometric mean diameter of approximately 50 nm and a total particle emission rate of 10(14) particles km(-1). Using 350 ppm high sulfur fuel and the standard oxidation catalyst a bimodal size distribution with a nucleation mode at 10 nm was observed at car velocities of 100 km h(-1) and 120 km h(-1), respectively. Nucleation mode particles were only present if high sulfur fuel was used with the oxidation catalyst installed. This is in agreement with prior work that these particles are of semivolatile nature and originate from the nucleation of sulfates formed inside the catalyst. Temporal effects of the occurrence of nucleation mode particles during steady state cruising and the dynamical behavior during acceleration and deceleration were investigated. PMID- 14524438 TI - Effect of simulated rainfall and weathering on release of preservative elements from CCA treated wood. AB - The release of arsenic from wood pressure-treated with chromated copper arsenate (CCA) can be decreased by application of wood finishes, but little is known about the types of finishes that are best suited for this purpose. This study evaluated the effects of finish water repellent content and ultraviolet (UV) radiation on the release of arsenic, copper, and chromium from CCA-treated wood exposed to simulated rainfall. Deck boards treated with CCA were either left unfinished or dipped in a finish prepared with 1%, 3%, or 5% water repellent. All specimens were exposed to leaching from simulated rainfall, and a subset of specimens was also exposed to UV radiation. The rainfall was collected and analyzed for total elemental arsenic, copper, and chromium. The water repellent significantly decreased the amounts of these elements in the runoff, but for the short duration of this study there was no difference among the three water repellent concentrations. It is possible that water repellent content would have a greater effect over a longer exposure period. Exposure to UV radiation caused a significant increase in leaching from both finished and unfinished specimens. This effect may be a result of increased surface area during weathering as well as loss of fibers caused by UV-induced surface erosion. PMID- 14524439 TI - Arsenic speciation and reactivity in poultry litter. AB - Recent U.S. government action to lower the maximum concentration levels (MCL) of total arsenic (As) (10 ppb) in drinking water has raised serious concerns about the agricultural use of As-containing biosolids such as poultry litter (PL). In this study, solid-state chemical speciation, desorbability, and total levels of As in PL and long-term amended soils were investigated using novel synchrotron based probing techniques (microfocused (micro) synchrotron X-ray fluorescence (SXRF) and micro-X-ray absorption near-edge structure (XANES) spectroscopies) coupled with chemical digestion and batch experiments. The total As levels in the PL were as high as approximately 50 mg kg(-1), and As(II/III and V) was always concentrated in abundant needle-shaped microscopic particles (approximately 20 microm x 850 microm) associated with Ca, Cu, and Fe and to a lesser extent with S, Cl, and Zn. Post-edge XANES features of litter particles are dissimilar to those of the organo-As(V) compound in poultry feed (i.e., roxarsone), suggesting possible degradation/transformation of roxarsone in the litter and/or in poultry digestive tracts. The extent of As desorption from the litter increased with increasing time and pH from 4.5 to 7, but at most 15% of the total As was released after 5 d at pH 7, indicating the presence of insoluble phases and/or strongly retained soluble compounds. No significant As accumulation (< 15 mg kg( 1)) was found in long-term PL-amended agricultural surface soils. This suggests that As in the PL may have undergone surface and subsurface transport processes. Our research results raise concerns about long-term PL amendment effects on As contamination in surrounding soil-water environments. PMID- 14524440 TI - Localization and speciation of chromium in subterranean clover using XRF, XANES, and EPR spectroscopy. AB - Optimization of phytoremediation and assessment of potential health hazards from metals in the environment requires an understanding of absorption, localization, and transport of the target metal by plants. The objectives of this study were to localize Cr and determine the oxidation state and possible complexation mode of Cr in intact plant tissue by means of XANES, synchrotron XRF microprobe spectroscopy, and EPR spectroscopy. Subterranean clover (Trifolium brachycalycinum) was grown hydroponically with Cr(VI) (0.04-2.0 mmol L(-1)) and compared with plants grown without Cr and with inorganic Cr(III) and various Cr(III)-organic sources. The uptake, translocation, and form of Cr in the plant were dependent on the form and concentration of supplied Cr. Chromium was found predominately in the +3 oxidation state, regardless of the Cr source supplied to the plant, though at high Cr(VI) treatment concentrations, Cr(VI) and Cr(V) were also observed. At low Cr(VI) concentrations, the plant effectively reduced the toxic Cr(VI) to less toxic Cr(III), which was observed both as a Cr(III) hydroxide phase at the roots and as a Cr(III)-organic complex in the roots and shoots. At low Cr(VI) treatment concentrations, Cr in the leaves was observed predominately around the leaf margins, while at higher concentrations Cr was accumulated at leaf veins. PMID- 14524441 TI - Sorption and dissipation of testosterone, estrogens, and their primary transformation products in soils and sediment. AB - Concern over the potential negative ecological effects of steroid hormones from human- and animal-derived wastes has resulted in an increased interest regarding the mobility and persistence of these compounds in the environment. Batch experiments were conducted to examine the simultaneous sorption and dissipation of three reproductive hormones (testosterone, 17beta-estradiol, and 17alpha ethynyl estradiol) in four midwestern U.S. soils and one freshwater sediment. Sorption isotherms were generated by measuring aqueous concentrations and by extracting the sorbed parent chemical or transformation products (e.g., estrone, androstenedione). Apparent sorption equilibrium is reached within a few hours. Measured sorption isotherms for the three parent chemicals and their principal transformation products were generally linear. Average organic carbon normalized sorption coefficients (K(oc)) resulted in standard deviations of less than 0.2 log units and were consistent with reported aqueous solubilites and octanol-water partition coefficients, indicating hydrophobic partitioning as the dominant sorption mechanism. Large log K(oc) values (approximately 3-4) suggest that leaching from soils will be limited, runoff of soil- and land-applied biosolids are the most likely inputs into surface waters, and that a significant fraction of these compounds will be associated with sediments. Half-lives for hormone dissipation in the aerobic soil and sediment slurries estimated assuming pseudo first-order processes ranged from a few hours to a few days with testosterone having the shortest half-life. PMID- 14524442 TI - Cd adsorption properties of components in different freshwater surface coatings: the important role of ferromanganese oxides. AB - Surface coatings developed in different natural waters were used to study the role of the composition of surface coatings in controlling Cd adsorption in aquatic environments. To investigate the adsorption property of each component, the method of extraction techniques followed by Cd adsorption and statistical analysis were employed. Hydroxylamine hydrochloride was used to remove Mn oxides selectively, sodium dithionite was used to remove Mn and Fe oxides, and oxalic acid was used to remove most metal oxides and part of the organic material. Adsorption of Cd to surface coatings was measured before and after extraction under controlled laboratory conditions. The observed Cd adsorptions to unextracted and extracted surface coatings were analyzed using nonlinear least squares fitting to estimate the adsorption property of each surface coating constituent. In different waters, the relative contribution to Cd adsorption of each component was different, but in all the waters studied, ferromanganese oxides contributed most with lesser roles indicated for organic phase and Al oxides. The Cd adsorption ability of manganese oxides was significantly higher than that of the other components. PMID- 14524443 TI - Gas-particle partitioning of semivolatile organic compounds (SOCs) on mixtures of aerosols in a smog chamber. AB - The partitioning behavior of a set of diverse SOCs on two and three component mixtures of aerosols from different sources was studied using smog chamber experimental data. A set of SOCs of different compound types was introduced into a system containing a mixture of aerosols from two or more sources. Gas and particle samples were taken using a filter-filter-denuder sampling system, and a partitioning coefficient Kp was estimated using Kp = Cp/(CgTSP). Particle size distributions were measured using a differential mobility analyzer and a light scattering detector. Gas and particle samples were analyzed using GCMS. The aerosol composition in the chamber was tracked chemically using a combination of signature compounds and the organic matter mass fraction (f(om)) of the individual aerosol sources. The physical nature of the aerosol mixture in the chamber was determined using particle size distributions, and an aggregate Kp was estimated from theoretically calculated Kp on the individual sources. Model fits for Kp showed that when the mixture involved primary sources of aerosol, the aggregate Kp of the mixture could be successfully modeled as an external mixture of the Kp on the individual aerosols. There were significant differences observed for some SOCs between modeling the system as an external and as an internal mixture. However, when one of the aerosol sources was secondary, the aggregate model Kp required incorporation of the secondary aerosol products on the preexisting aerosol for adequate model fits. Modeling such a system as an external mixture grossly overpredicted the Kp of alkanes in the mixture. Indirect evidence of heterogeneous, acid-catalyzed reactions in the particle phase was also seen, leading to a significant increase in the polarity of the resulting aerosol mix and a resulting decrease in the observed Kp of alkanes in the chamber. The model was partly consistent with this decrease but could not completely explain the reduction in Kp because of insufficient knowledge of the secondary organic aerosol composition. PMID- 14524444 TI - Sorption and manganese-induced oxidative coupling of hydroxylated aromatic compounds by natural geosorbents. AB - The sorption/desorption equilibria and solvent extractabilities of phenol, o cresol, and p-chlorophenol with respect to natural sorbents having different types of soil organic matter were investigated. Parallel tests in systems amended with birnessite (delta-MnO2), a solid-phase oxidative coupling catalyst, were also conducted. Sorption/desorption isotherms and solvent extraction data reveal that the relative isotherm linearities, desorption hysteresis, and extractabilities of these compounds are related to the geochemical nature of the sorbent organic matter and to the existence of system conditions that promote oxidative coupling reactions. When suitable coupling catalysts are present, soils containing primarily diagenetically "young" and highly amorphous organic matter (e.g., humic materials) are more likely to retain those solutes than are those containing primarily diagenetically "old" and more condensed organic matter (e.g., kerogens). The sorption/desorption properties of the solutes were significantly altered in the presence of birnessite as a result of both cross coupling reactions with reactive soil organic matter components and self-coupling reactions with each other to form polymeric species. Under appropriate conditions, mineral-catalyzed oxidative coupling may exert a dominant influence on the sorption and transport of hydroxylated aromatic compounds in soil and sediment systems. PMID- 14524445 TI - Dissolution of entrapped DNAPLs in variable aperture fractures: experimental data and empirical model. AB - An appreciation of the dissolution from entrapped nonaqueous phase liquids (NAPLs) in fractures is essential as we attempt to understand and predict the fate of NAPLs present in fractured rock systems. Eight long-term dissolution experiments using 1,1,1-trichloroethane and trichloroethylene were conducted in two laboratory-scale dolomitic limestone variable aperture fractures under various conditions. Between 560 and 2600 fracture volumes of water were passed through the fractures resulting in the removal of 10-60% of the initial mass trapped. The effluent concentration profiles revealed three distinct and characteristic stages of dissolution: an initial pseudosteady stage, a transient stage, and a tailing stage. On average, 8% of the initial volume of NAPL present was removed during the initial pseudosteady stage. Data from the dissolution experiments were used in conjunction with statistical techniques to develop a continuous empirical model describing the initial pseudosteady and transient stages of dissolution. The model was used to successfully replicate effluent concentration data from two separate and independent dissolution experiments. The experimental results provide an indication of the expected dissolution behavior of entrapped NAPLs, while the developed model is a useful tool for characterizing mass transfer rates in variable aperture fractures. PMID- 14524447 TI - Assessing influence of experimental parameters on formation of PCDD/F from ash derived from fires of CCA-treated wood. AB - Ash residues from fires of radiata pine timber, both untreated and treated with chromated copper arsenate (CCA), were analyzed for the presence of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/F). Fire conditions were simulated using a cone calorimeter. The sensitivity of the magnitude and profile of PCDD/F in the ash under controlled experimental conditions were examined to gain an insight into the formation of PCDD/F in a system containing CCA. The total amount of PCDD/F increased from 2.0 ng/kg of ash (0.05 ng of TE/kg of ash, using WHO-TEF) for untreated radiata pine to a maximum of 2700 ng/kg of ash (78 ng of TE/kg of ash) for 0.94% CCA. Ash containing CCA showed a distinct preference for formation of PCDFs, particularly the tetrachloro homologue. It is concluded that PCDD/F formation predominantly occurred via de novo synthesis during smoldering of the char rather than during the initial flaming and pyrolysis. Furthermore, the composition of the CCA constituents present in the timber was controlled to assess whether the physical presence of Cu, a known catalyst in PCDD/F production, was sufficient to account for the formation of PCDD/F in fires of timber impregnated with CCA. PMID- 14524448 TI - Influence of autochthonous dissolved organic carbon and nutrient limitation on alachlor biotransformation in aerobic aquatic systems. AB - Much work has suggested that the rate of attenuation of water-soluble organic contaminants in aerobic aquatic systems is dependent on the level of secondary nutrients in the water column. For example, the decay rate of alachlor, a common herbicide, was over 10 times higher under hypereutrophic compared with oligotrophic water conditions. It has been presumed that higher water column nutrient levels produce larger microbial communities, resulting in higher rates of alachlor cometabolism. However, most earlier field studies only assessed alachlor fate in systems with full light exposure (FLE). Therefore, new experiments were performed to assess how variations in light level affect alachlor cometabolism in such systems. Twelve tank mesocosms were maintained using identical nitrogen (N) and phosphorus (P) supply conditions: four units with full light exposure (100% FLE), four with partial shading (19.3% FLE), and four with near complete shading (0.5% FLE). Alachlor half-lives were found to vary broadly, from 50 to 60 days in higher light units to > 180 days in the 0.5% FLE units. Nutrient analysis indicated that the low light units were severely carbon (C)-limited for microbial decomposition, whereas the other units had excess C relative to N and P. Apparently, reduced light levels cause decreased production of bioavailable C for decomposition, which significantly reduces alachlor cometabolism. The data suggest that water column nutrient levels only correlate with the alachlor decay rate when light levels are high, and that the biodegradable carbon supply must be considered when the fate of water-soluble contaminants in aerobic aquatic systems is assessed. PMID- 14524446 TI - Biofilm hydrous manganese oxyhydroxides and metal dynamics in acid rock drainage. AB - Biofilms in shallow, tailings-associated acid rock drainage (ARD) accumulated metals from May to September, indicating scavenging is stable within these biological solids over seasonal time frames. Results indicate a doubling (Mn, Cr) to over a 6-fold increase (Ni, Co) in biofilm metal concentrations. Biofilm oxygen and pH gradients measured over diel time scales with microelectrodes were observed to be both spatially and temporally variable, indicating that biofilms are highly dynamic geochemical environments. Biofilm metal retention and affinities were element specific indicating different processes control their sequestration. Metals were specifically scavenged by the organic constituents of the biofilm itself (Ni, Co) and associated biominerals of amorphous Mn oxyhydroxides (HMO; Ni, Co, and Cr). Results are consistent with sorption and coprecipitation processes controlling Ni and Co biofilm association, while Cr dynamics appear linked to those of Mn through redox processes. Biofilm HMO concentrations increased seasonally but showed significant diel fluctuations, indicating that both formation and dissolution processes occurred over rapid time scales in these biofilms. Biofilm HMO concentrations increased nocturnally but decreased during daylight hours to late afternoon minima. Under the geochemical conditions of the streams, observed HMO formation rates can only be explained by microbial catalysis. These results are the first to quantitatively examine microbial biofilm metal dynamics using microscale, geochemical techniques at both diel and seasonal time scales. They provide strong evidence for the significant role that microbial activity can play in metal geochemistry in natural environments. PMID- 14524449 TI - Spectroscopic studies of the biosorption of gold(III) by dealginated seaweed waste. AB - Gold biosorption by dealginated seaweed waste has been studied to elucidate the mechanisms of metal uptake from solution. Dealginated seaweed was able to retain up to 1 mmol g(-1) of Au from solution at pH 3. FT-IR showed the presence of carboxylate groups on the surface of the biosorbent; however, the changes observed for the Au-bound samples suggested very little sorption to the carboxyl moieties. Colloidal Au formed on the surface of dealginated seaweed by reduction of Au(III) to Au(0) was observed using ESEM and four different types of particles were clearly identified. The Au distribution matched closely that obtained for S atoms indicating a possible link between these elements. EXAFS measurements showed that colloidal Au is present on the surface of the biosorbent. Evidence of gold reduction from Au(III) to Au(I) and Au(0) was also confirmed by the measured bond distances characteristic of the metal. The coordination number obtained by EXAFS indicated that approximately 75% of the Au on the sample was present in the colloidal form and the remaining Au was bound to S as nearest neighbor. The proposed mechanisms for Au removal from solution are reduction of Au species by components on the surface of the biosorbent to form colloidal metal followed by retention of the ionic Au(I) species at the sulfur containing sites. The results show that dealginated seaweed can be used for the cleanup of gold-containing effluents. PMID- 14524450 TI - The OH-initiated oxidation of hexylene glycol and diacetone alcohol. AB - The OH-initiated oxidation of two VOCs directly emitted to the atmosphere through their use as industrial solvents, hexylene glycol (HG, (CH3)2C(OH)CH2CH(OH)CH3) and diacetone alcohol (DA, (CH3)2C(OH)CH2C(O)CH3), has been studied in two photoreactors: a 140 L Teflon bag irradiated by lamps at CNRS-Orleans and the 200 m3 European photoreactor, EUPHORE, irradiated by sunlight. The rate constants for the reactions of HG and DA with OH radicals have been determined at (298 +/- 3) K using a relative rate method: k(HG) = (1.5 +/- 0.4) x 10(-11) and k(DA) = (3.6 +/ 0.6) x 10(-12) cm(3) molecule(-1) s(-1) and have been found in good agreement with estimations from structure-reactivity relationships. The study at Orleans and EUPHORE of the OH-initiated oxidation of hexylene glycol showed the formation of diacetone alcohol, acetone, and PAN as the principal products. The branching ratio of the H-atom abstraction from the > CH- group of HG has been estimated to be (47 +/- 4)% corresponding to the measured formation yield of DA. The formation yields of acetone and PAN lead to the determination of a lower limit of (33 +/- 7)% for the branching ratio of the H-atom abstraction of the -CH2- group of HG. For diacetone alcohol, studies at EUPHORE have shown negligible photolysis under atmospheric conditions (J < 5 x 10(-6) s(-1)) and the formation of acetone, PAN, HCHO, and CO in the OH-initiated oxidation experiments. The molar yield of acetone, close to 100%, corresponds to the branching ratio of the H-atom abstraction from the -CH2- group of DA. The present study has allowed the identification of the nature and the fate of the oxy radicals as intermediates in the oxidation mechanism of both HG and DA. The atmospheric implication of these results, especially the ozone formation potential of HG and DA, is discussed. PMID- 14524451 TI - Comparison of arsenic(V) and arsenic(III) sorption onto iron oxide minerals: implications for arsenic mobility. AB - Arsenic derived from natural sources occurs in groundwater in many countries, affecting the health of millions of people. The combined effects of As(V) reduction and diagenesis of iron oxide minerals on arsenic mobility are investigated in this study by comparing As(V) and As(III) sorption onto amorphous iron oxide (HFO), goethite, and magnetite at varying solution compositions. Experimental data are modeled with a diffuse double layer surface complexation model, and the extracted model parameters are used to examine the consistency of our results with those previously reported. Sorption of As(V) onto HFO and goethite is more favorable than that of As(III) below pH 5-6, whereas, above pH 7 8, As(II) has a higher affinity for the solids. The pH at which As(V) and As(III) are equally sorbed depends on the solid-to-solution ratio and type and specific surface area of the minerals and is shifted to lower pH values in the presence of phosphate, which competes for sorption sites. The sorption data indicate that, under most of the chemical conditions investigated in this study, reduction of As(V) in the presence of HFO or goethite would have only minor effects on or even decrease its mobility in the environment at near-neutral pH conditions. As(V) and As(III) sorption isotherms indicate similar surface site densities on the three oxides. Intrinsic surface complexation constants for As(V) are higher for goethite than HFO, whereas As(III) binding is similar for both of these oxides and also for magnetite. However, decrease in specific surface area and hence sorption site density that accompanies transformation of amorphous iron oxides to more crystalline phases could increase arsenic mobility. PMID- 14524452 TI - Photoformation of low-molecular-weight organic acids from brown water dissolved organic matter. AB - This work describes the effects of simulated solar UV light on the bulk properties of dissolved organic matter (DOM) of bog lake water and on the formation of low-molecular-weight organic acids (LMWOAs). By means of size exclusion chromatography it was shown that the more hydrophilic moieties of the DOM were preferentially photodegraded while the more hydrophobic ones remained relatively unaffected or were even formed. The combined photochemical-biological degradation proved to be more important than the pure photochemical mineralization. Formic, acetic, pyruvic, oxalic, malonic, and succinic acids were identified as important degradation products. Their contribution to the dissolved organic carbon increased from 0.31% before to 6.4% after 24 h irradiation. About 33% of the bioavailable photoproducts of DOM were comprised of these LMWOAs. The influence of nitrate on the formation of carboxylic acids could not be observed in the investigated system. Kinetic experiments indicated that degradation of LMWOAs occurred simultaneously during irradiation experiments, alpha-oxygen substituted LMWOAs being more amenable to these processes. Dissolved iron acted as a catalyst of DOM photodegradation and LMWOA photoformation. Copper played an antagonistic role in the irradiation experiments, reducing the formation of formic, acetic, and malonic acids while increasing the formation of oxalic acid. PMID- 14524453 TI - Theoretically predicted rate constants for mercury oxidation by hydrogen chloride in coal combustion flue gases. AB - In this work, theoretical rate constants are estimated for mercury oxidation reactions by hydrogen chloride that may occur in the flue gases of coal combustion. Rate constants are calculated using transition state theory at the quadratic configuration interaction (QCI) level of theory with single and double excitations, and are compared to results obtained from density functional theory, both including high level pseudopotentials for mercury. Thermodynamic and kinetic data from the literature are used to assess the accuracy of the theoretical calculations when possible. Validation of the chosen methods and basis sets is based upon previous and current research on mercury reactions involving chlorine. The present research shows that the QCISD method with the 1992 Stevens et al. basis set leads to the most accurate kinetic and thermodynamic results for the oxidation of mercury via chlorine containing molecules. Also, a comparison of the heats of reaction data for a series of mercury oxidation reactions reveals that the density functional method, B3LYP, with the 1997 Stuttgart basis set provides reasonably accurate results for these large systems. PMID- 14524454 TI - Isotopic and geochemical assessment of in situ biodegradation of chlorinated hydrocarbons. AB - Currently there is no in situ method to detect and quantify complete mineralization of chlorinated hydrocarbons (CHCs) to CO2. Combined isotopic measurements in conjunction with traditional chemical techniques were used to assess in situ biodegradation of trichloroethylene (TCE) and carbon tetrachloride (CT). Vadose zone CHC, ethene, ethane, methane, O2, and CO2 concentrations were analyzed using gas chromatography over 114 days at the Savannah River Site. delta13C of CHC and delta13C and 14C of vadose zone CO2, sediment organic matter, and groundwater dissolved inorganic carbon (DIC)were measured. Intermediate metabolites of TCE and CT accounted for < or = 10% of total CHCs. Delta13C of cis 1,2-dichloroethylene (DCE) was always heavier than TCE indicating substantial DCE biodegradation. 14C-CO2 values ranged from 84 to 128 percent modern carbon (pMC), suggesting that plant root-respired CO2 was dominant. 14C-CO2 values decreased over time (up to 12 pMC), and contaminated groundwater 14C-DIC (76 pMC) was substantially depleted relative to the control (121 pMC). 14C provided a direct measure of complete CHC mineralization in vadose zone and groundwater in situ and may improve remediation strategies. PMID- 14524455 TI - Dissolved PCBs, PAHs, and HCB in pore waters and overlying waters of contaminated harbor sediments. AB - Aqueous concentrations of polychlorinated biphenyls (PCBs), polyaromatic hydrocarbons (PAHs), pentachlorobenzene, and hexachlorobenzene (HCB) were determined by exposing low-density polyethylene (LDPE) strips to the pore waters and the overlying water in two contaminated harbors. LDPE strips were also exposed in the laboratory to sediment slurries and to stagnant sediments collected at the same locations. Surprisingly short equilibration time scales (1 60 days for log K(ow) < 7) were observed for the exposures to sediment slurries. This was a result of a profound decrease in transport resistance, due to the presence of particles in the aqueous boundary layer. Concentrations in the pore water were calculated from the initial uptake rates and the dissipation rates of performance reference compounds. Good correspondence existed with concentrations estimated from the equilibrium amounts in the strips and LDPE-water partition coefficients. Sediment-water partition coefficients for PAHs were higher than for PCBs and chlorobenzenes by approximately 1 order of magnitude. A one-dimensional diffusion model was used to describe contaminant uptake by LDPE strips from stagnant sediments. The results indicated that 95% of the PAHs and 50% of the PCBs were immobile on a time scale of two months. A comparison of concentrations in pore waters and water columns indicated that a fair degree of equilibrium existed for PCBs and that one sediment was a potential source of PAHs. Concentrations of HCB near a former discharge site were higher by a factor of 6 compared to reference locations. PMID- 14524456 TI - Inclusion of persistent organic pollutants in humification processes: direct chemical incorporation of phenanthrene via oxidative coupling. AB - The participation of phenanthrene in phenol-coupling reactions mediated by horseradish peroxidase was investigated. Aqueous-phase concentrations of phenanthrene were observed to decrease dramatically with phenol as a result of the formation of precipitated products, suggesting a potential means for simultaneous treatment of phenolic contaminants and polycyclic aromatic hydrocarbons (PAHs) using peroxidase-mediated oxidative coupling processes. The studies reveal that phenanthrene removal from the aqueous phase occurs by a combination of sorption by and chemical bonding to precipitated reaction products. In that the oxidative coupling reactions of phenolics comprise an important step in the initiation of humification processes, the results obtained provide insights to potential roles that natural humification may play in the sorption, sequestration, and environmental fate of PAHs and other organic xenobiotics of similar nature. PMID- 14524457 TI - Dimethyl sulfide removal from synthetic waste gas using a flat poly(dimethylsiloxane)-coated composite membrane bioreactor. AB - The reduction of volatile organic sulfur emissions should be completely as they cause odor nuisance, even when they are emitted in very small amounts. In general, biofilters are applied for odor reduction, but their operational control is limited. A new biotechnique for the treatment of complex emissions is the use of membranes integrated within bioreactors (MBRs). In this study, the reactor performance of MBRs for removal of dimethyl sulfide (DMS) as a model compound is presented. Composite membranes with a poly(dimethylsiloxane) (PDMS)-coating layer were used. The MBRs were inoculated with the sulfur-degrading culture Hyphomicrobium VS or a suspension of Hyphomicrobium VS, ammonium-oxidizing bacteria (AOB), and nitrite-oxidizing bacteria (NOB) to colonize the PDMS-coating layer. Although inoculation with AOB and NOB might give rise to competition for space on the membrane, their presence in the MBRs appeared to be positive as they co-oxidize DMS. Dimethyl sulfide elimination depended on the inoculum type, DMS inlet concentration, gas residence time, and membrane polymer. For equal loading rates, the elimination capacity (EC) increased at larger gas residence times and inlet concentrations. The maximum EC obtained with the MBRs was 4.8 kg of DMS x m(-3) x d(-1). This value is higher than any reported figure for biofilters and biotrickling filters. PMID- 14524458 TI - Removal of hexavalent chromium with a lignocellulosic substrate extracted from wheat bran. AB - In this paper, a new recovery system of the toxic hexavalent chromium Cr(VI) is proposed that uses a lignocellulosic substrate derived from the industrial treatment process of wheat bran. We studied the adsorption mechanism of Cr(VI) onto the lignocellulosic substrate and showed that the adsorption reaction consumes a large amount of protons goes along the reduction of Cr(VI) into Cr(III). The oxidation of lignin moieties takes place concurrently to the chromium reduction and leads to the formation of hydroxyl and carboxyl functions. The latter contribute to an increase in the number of ion-exchange sites for the reduced chromium. The maximum adsorption capacity for hexavalent chromium was found at about 35 mg g(-1) in an acidic medium. This is comparable to other natural substrates and ordinary adsorbents. PMID- 14524459 TI - Formation of methyl nitrite and methyl nitrate during plasma treatment of diesel exhaust. AB - FTIR spectroscopy was used to identify CH3ONO and CH3ONO2 as products of the nonthermal plasma treatment of simulated diesel exhaust. This is the first observation of CH3ONO formation in such systems. The yield of CH3ONO relativeto CH3ONO2 scaled linearly with the average [NO]/ [NO2] ratio in the system. A plot of [CH3ONO]/[CH3ONO2] versus [NO]/[NO2] gives a slope of 1.81 +/- 0.30. This result is indistinguishable from the literature value of the rate constant ratio k(CH3O + NO)/k(CH3O + NO2) = (2.6 x 10(-11))/ (1.5 x 10(-11)) = 1.73 +/- 0.37. The experimental observations suggest that reactions of CH3O radicals with NO and NO2 are the sources of CH3ONO and CH3ONO2 in such systems. The linear relationship between the yields of CH3ONO and CH3ONO2 provides a means of estimating the yield of these compounds during nonthermal plasma treatment of diesel exhaust. PMID- 14524460 TI - Use of a surfactant-stabilized emulsion to deliver 1-butanol for density-modified displacement of trichloroethene. AB - A novel surfactant-enhanced aquifer remediation technology, density-modified displacement (DMD), has been developed to minimize risk of dense non-aqueous phase liquid (DNAPL) downward migration during displacement floods. The DMD method is designed to be implemented using horizontal flushing schemes, with in situ DNAPL density conversion accomplished by the introduction of a partitioning alcohol (e.g., 1-butanol) in a predisplacement flood (preflood). Subsequent NAPL displacement and recovery is achieved by flushing with a low-interfacial-tension (low-IFT) surfactant solution. The efficiency of the DMD method may be enhanced for heavier DNAPLs, such as trichloroethene (TCE), by increasing alcohol delivery and the extent of partitioning during the preflood. The objective of this study was to evaluate the use of a macroemulsion, consisting of 4.7% (vol) Tween 80 + 1.3% (vol) Span 80 + 15% (vol) 1-butanol to achieve efficient in situ density conversion of TCE (relative to that obtained with use of an aqueous preflood solution) prior to low-IFT displacement and recovery from a two-dimensional aquifer cell. The cell was configured to represent a heterogeneous unconfined aquifer system with an overall NAPL saturation between 2% and 3%. After flooding with approximately 1.2 pore volumes of the macroemulsion, a low-IFT solution consisting of 10% (vol) Aerosol MA + 6% (vol) 1-butanol + 15 g/L NaCl + 1 g/L CaCl2 was introduced to displace and recover NAPL. Visual observations and quantitative measurements of effluent fluids demonstrated that in situ density conversion and displacement of TCE-NAPL was successful, with effluent NAPL densities ranging from 0.97 to 0.99 g/mL. For the experimental system employed herein, 93% recovery of the introduced TCE mass was realized after flushing with a combined 2.4 pore volumes of the density conversion and low-IFT solutions. These results demonstrate the increased efficiency of the DMD method when surfactant-based emulsions are used to enhance 1-butanol delivery and partitioning behavior. PMID- 14524461 TI - Development of supported iron oxide catalyst for destruction of PCDD/F. AB - Studies on the development of supported iron oxide catalysts for PCDD/F decomposition using 2-monochlorophenol as a surrogate test compound are presented. Iron oxide catalysts supported on titania were prepared by two methods: impregnation and the sol-gel method. The latter preparation method resulted in better dispersion of iron oxide on the surface and the formation of gamma-Fe2O3. This is in contrast to the impregnated samples where alpha-Fe2O3 crystallites were formed. Formation of gamma-Fe2O3 resulted in improved reducibility of the active phase that favorably affected the catalytic oxidation properties of the catalyst, i.e., the light-off curves for the sol-gel samples were shifted toward lower temperature. Addition of calcium oxide to iron oxide catalyst further improved the performance of the system through stabilization and increase in the concentration of gamma-Fe2O3 in the sol-gel prepared samples. Addition of calcium oxide has a dual effect on the performance of the catalyst. First, it creates oxygen vacancies in the reduction-resistant Fe2O3 octahedral structures, thereby improving the reducibility of the active phase. Second, iron oxide can transform during decomposition of chlorinated hydrocarbons into iron chloride. Calcium oxide improved the chlorine transfer from the surface iron oxide species, thereby providing a relatively fresh surface for further catalytic oxidation. Comparison of TPR profiles with the position of light-off curves in 2 monochlorophenol decomposition led to the conclusion that Fe3O4 species are the active phase under conditions that facilitate redox cycling between Fe3+ and Fe2+ ions. PMID- 14524462 TI - Novel polymeric chelating fibers for selective removal of mercury and cesium from water. AB - We report here the synthesis and characterization of two new classes of chelating fibers, namely, (1) polymercaptopropylsilsesquioxane (PMPS) and (2) copper(II) ferrocyanide complexed with poly[1-(2-aminoethyl)-3-aminopropyl]silsesquioxane (Cu-FC-PAEAPS) fibers. These fibers were evaluated for selective removal of trace amount of mercury and cesium ions respectively in the presence of competing metal ions from water. The PMPS and Cu-FC-PAEAPS fibers were prepared by coating their corresponding soluble prepolymers, which are derived from mercaptopropyltrimethoxysilane and [1-(2-aminoethyl)-3 aminopropyl]trimethoxysilane monomers, respectively, on a glass fiber substrate, followed by a cross-linking step at 120 degrees C. The fibers were characterized through infrared spectroscopy, scanning electron microscopy (SEM), and thermogravimetric analysis (TGA). These novel materials are extremely efficient in removing low concentrations of mercury and cesium ions from water in the presence of high concentrations of sodium or potassium ions. They were shown to remove trace mercury and cesium contaminants effectively to well below parts per billion concentrations under a variety of conditions. PMID- 14524463 TI - Use of copper shavings to remove mercury from contaminated groundwater or wastewater by amalgamation. AB - The efficacy of copper shavings (Cu(0)) for the removal of Hg2+ from aqueous solution by amalgamation is demonstrated. Two kinds of copper shavings were investigated: (a) chemically processed shavings (Fluka) and (b) recycled shavings from scrap metal. Batch sorption experiments yielded very high retardation coefficients of 28 850-82 830 for the concentration range studied (1-10 000 microg/L Hg2+ dissolved in distilled water or in a 0.01 M CaCl2 matrix solution). Sorption data were well-described bythe Freundlich isotherm equation. Kinetic batch sorption experiments showed that 96-98% of Hg2+ was removed within 2 h. Column experiments were performed with a mercury solution containing 1000 microg/L Hg in a 0.01 M CaCl2 matrix with a flow rate of 0.5 m/d. No mercury breakthrough (c/c(0) = 0.5) could be detected after more than 2300 percolated pore volumes, and the high retardation coefficients determined in the batch studies could be confirmed. Copper was released from the shavings due to the amalgamation process and to copper corrosion by oxygen, resulting in concentrations of mobilized copper of 0.2-0.6 mg/L. Due to their high efficiency in removing Hg2+ from aqueous solution, the use of copper shavings for the removal of mercury from contaminated water is suggested, employing a sequential system of mercury amalgamation followed by the removal of mobilized copper by an ion exchanger such as zeolites. Possible applications could be in environmental technologies such as wastewater treatment or permeable reactive barriers for in situ groundwater remediation. PMID- 14524464 TI - Patchy biofilm coverage can explain the potential advantage of BGAC reactors. AB - An adsorbing biofilm carrier, like granular activated carbon (GAC), can be the source of an extra flux of pollutant to the biofilm in addition to the bulk liquid. This double flux can improve the performance of a biological GAC (BGAC) reactor as compared to a nonabsorbing carrier reactor but only under conditions of pollutant partial penetration in the biofilm. Pollutant partial penetration in a biofilm often occurs in treatment processes where very low effluent concentrations are required. However, under these conditions, adsorption in BGAC reactors is questionable and requires the existence of biofilm free areas on the GAC carrier. The purpose of this investigation is to prove that under normal BGAC fluidized bed reactor operational conditions patchy biofilm coverage with exposed areas of GAC develops. Adsorption and desorption through these exposed areas can explain the widely debated advantage of BGAC reactors regarding higher biofilm activity. The patchy-like nature of the biofilm coverage on the GAC particles was verified using experimental and modeling tools. Comparison between a nonadsorbing granular carbon carrier and a GAC carrier with an atrazine degrading biofilm (Pseudomonas ADP) under conditions of atrazine partial penetration in the biofilm showed higher biodegradation and lower effluent atrazine concentrations in the BGAC reactor. PMID- 14524465 TI - Co-removal of hexavalent chromium through copper precipitation in synthetic wastewater. AB - The mechanisms of hexavalent chromium [Cr(VI)] co-removal with copper [Cu(II)] during homogeneous precipitation were studied with batch tests using a synthetic solution containing Cr(VI) and Cu(II). Metal precipitation was induced by adding Na2CO3 stepwise to different pH, and the respective removals of Cu(II) and Cr(VI) were measured. At the same time, the relative quantities of Cu(II) and Cr(VI) in the precipitates were also analyzed to establish their stoichiometric relationship. The results indicated that, in a solution containing 150 mg/L Cu(II) and 60 mg/L Cr(VI), the initial co-removal of Cr(VII with Cu(II) began at pH 5.0 and completed at pH 6.2. At pH 5.0-5.2, coprecipitation took place through the formation of copper-chromium-bearing solids [such as CuCrO4 and/or CuCrO4 x 2Cu(OH)2]. Thereafter, the remaining soluble copper started to react with carbonate in a heterogeneous environment to form the negatively charged basic copper carbonate precipitates [CuCO3 x Cu(OH)2], which subsequently adsorbed additional Cr(VI) (or HCrO4-) at pH 5.2-6.2. The maximum Cr(VI) co-removal took place at pH 6.2. Between the two mechanisms, co-precipitation accounted for about 29% of the total chromium's co-removal while the remaining 71% was attributed to surface adsorption, mainly through electrostatic attraction and ligand exchange. When the solution pH was increased to beyond 7.5, a surface charge reversal took place on the basic copper carbonate solids, and this led to some Cr(VI) desorption. Thus, the extent of Cr(VI) adsorption is highly pH dependent. PMID- 14524466 TI - Influence of media characteristics on energy dissipation in filter backwashing. AB - Effective cleaning of granular filters during backwashing processes needs maximum turbulence and maximum shear in the fluid particle field. The energy dissipation in a backwashed filter as a particulate fluidized bed arises due to the suspending and random motions of particles and turbulent fluctuations in the bed. Size, density, and sphericity of the filter materials greatly influence the fluidization behavior of the media. In this study, a new model is proposed for predicting the energy dissipation parameters namely the hydrodynamic shear stress (tau(a)), the velocity gradient (G(a)), the turbulence dissipation coefficient (C(a)), and the turbulence parameter (C(a)0.5/Re) in backwashing of filters for different types of filter materials (sand, anthracite, and glass ball). The hydrodynamic shear stress is the dominant mechanism of filter cleaning and appears to increase with increasing the density and size of the filter media particles. Using the basic set of data, a step by step procedure is developed to compute the velocity gradient G(a), the turbulence dissipation coefficient C(a), the hydrodynamic shear stress tau(a), and the turbulent parameter (C(a)0.5/ Re). PMID- 14524467 TI - Monitoring of landfill leachate dispersion using reflectance spectroscopy and ground-penetrating radar. AB - The utility of ground-penetrating radar and reflectance spectroscopy in the monitoring of landfill sites has been investigated. Strong correlations between red edge inflection position and chlorophyll and heavy metal concentrations have been demonstrated from grassland species affected by leachate contamination of the soil adjacent to the landfill test site. This study demonstrated that reflectance spectroscopy can identify vegetation affected by leachate contaminated soil at a range of spatial resolutions. To identify the vegetation affected by leachate contamination, the spectroradiometer must have contiguous bands at sufficient spectral resolution over the critical wave range that measures chlorophyll absorption and the red edge (between 650 and 750 nm). The utility of ground-penetrating radar data to identify leachate escaping from breakout points in the contaminant wall has also been demonstrated. An integrated approach using these techniques, combined with field and borehole sampling and contaminant migration modeling, offers a possible cost-effective monitoring approach for landfill sites. PMID- 14524468 TI - Biofiltration at composting facilities: effectiveness for bioaerosol control. AB - Biofiltration was evaluated as a method to control the airborne microorganisms released at composting facilities. Seven commercial composting plants were selected for this study because of their different operating conditions and biofilter designs. In all plants, the biofilters were originally designed for odor control. The concentrations of both Aspergillus fumigatus and mesophilic bacteria were measured in the air stream before and after passing through the biofilters and compared with the background concentrations in the surrounding area. Results showed that biofiltration achieved an average reduction greater than 90% and 39% in the concentrations of A. fumigatus and mesophilic bacteria, respectively. In all the plants, the airborne A. fumigatus concentration after the biofilter was lower than 1.2 x 10(3) cfu m(-3), independent of the inlet concentration, whereas the mesophilic bacteria concentration was dependent on the inlet concentration. The different behaviors of the two microorganism groups were thought to be due to the different aerodynamic characteristics of the particles that affected the capture by impact in the biofilter bed. The fungus, whose spores had a maximum of diameter size distribution between 2.1 and 3.3 microm, were more effectively captured in the biofilter than the bacteria, which had diameters mainly between 1.1 and 2.1 microm. PMID- 14524469 TI - Mitigating uranium in groundwater: prospects and limitations. AB - Removal of uranium(VI) by zerovalent iron has been suggested as a feasible pathway to control uranium contaminations in seepage waters. Available information in the literature however presents discrepant evidence on the process responsible for the mitigation effect. On basis of an EH-pH diagram of uranium and iron, it is outlined that these discrepancies may be explained by the aqueous chemistry of uranium and iron. Additional effects contributing to the complexity of the system are given. Solubilization experiments using scrap iron together with water works sludge, MnO2, and pyrite indicate that U(VI) is immobilized by iron corrosion products after about 50 days. PMID- 14524470 TI - Effect of truck operating weight on heavy-duty diesel emissions. AB - Heavy-duty diesel vehicles are substantial contributors of oxides of nitrogen (NO(x)) and particulate matter (PM) while carbon monoxide and hydrocarbon (HC) emissions from diesel vehicles receive less attention. Truck emissions inventories have traditionally employed average fuel economy and engine efficiency factors to translate certification into distance-specific (g/mi) data, so that inventories do not take into account the real effects of truck operating weight on emissions. The objective of this research was to examine weight corrections for class 7 and 8 vehicles (over 26 000 lb (11 793 kg) gross vehicle weight) from a theoretical point of view and to present a collection of original data on the topic. It was found by combining an empirical equation with theoretical truck loads that the NO(x) emissions increased by approximately 54% for a doubling of test weight. Emissions data were gathered from specific tests performed using different test weights and using various test schedules, which can consist of cycles or routes. It was found experimentally that NO(x) emissions have a nearly linear correlation with vehicle weight and did not vary much from vehicle to vehicle. NO(x) emissions were also found to be insensitive to transient operation in the test schedule. The observed trends correlate well with the theory presented, and hence, the NO(x) emissions can be predicted reasonably accurately using the theory. If NO(x) data were considered in fuel-specific (g/gal) units, they did not vary with the test weight. HC emissions were found to be insensitive to the vehicle weight. CO and PM emissions were found to be a strong function of weight during transient operation. Under transient operation, the CO emissions value increased by 36% for an increase in test weight from 42 000 (19 051 kg) to 56 000 lb (25 401 kg). However, CO and PM were found to be insensitive to the vehicle weight during nearly steady-state operation. PMID- 14524471 TI - Chloride concentration discriminates between Foot-and-mouth disease virus ires dependent translation and classical scanning translation: new aspects of the picornavirus shutoff mechanism. AB - Some picornaviruses might use the general increase of ionic strength in the host cell that occurs successively after infection to induce shutoff of host protein synthesis and to stimulate viral protein synthesis. In order to investigate this discrimination mode on a molecular level, in vitro experiments under different salt conditions comparing the Foot-and-mouth disease virus (FMDV) internal ribosome entry site (IRES)-dependent translation with the translation via the classical scanning mechanism were performed. For classical mRNA optimum concentrations of all investigated salts ranged between 70 and 100 mmol/l. However, for FMDV IRES-dependent translation the optima depended strongly on the anion used. While acetates caused only a weak stimulation of translation efficiency with maxima ranging between 150 and 180 mmol/l, chlorides lead to a strong stimulation with maxima ranging between 120 and 150 mmol/l. Competition experiments revealed that the concentration of chlorides had a greater influence on the discrimination between cellular and viral RNA translation than the total ionic strength. Taken together, the data support a model in which a specific increase in the chloride concentration rather than a general increase in the ionic strength is responsible for the shutoff effect induced by some picornaviruses. PMID- 14524472 TI - Sequence analysis of segment a of a field virus isolate from an outbreak of Infectious bursal disease in India. AB - Sequence analysis of genome segment A of an Indian Infectious bursal disease virus (IBDV) field isolate (KT1/99) revealed total 95 nucleotide substitutions, resulting in 17 amino acid changes. Of these, five amino acid changes, namely F60S, T137I, I374V, V519I and E682D were unique to the KT1/99 isolate. The amino acid change P222A and the proposed hot mutation spot 680Y, reported to be present in very virulent IBDV isolates were also found in KT1/99. This isolate had nucleotide divergence of 1.1% to 4.95% from the other reported serotype 1 IBDV isolates and 19.6% from serotype 2 strain OH in polyprotein gene sequence, while divergence at amino acid level was 0.6% to 2.9% and 11.4%, respectively. Based on both nucleotide and amino acid sequence analysis, KT1/99 was grouped phylogenetically with the reported Bangladesh isolate BD3/99 in one cluster along with other reported very virulent isolates in same lineage. PMID- 14524473 TI - RNA-dependent RNA polymerase activity of Classical swine fever virus NS5B protein expressed in natural host cells. AB - The NS5B gene, cloned from Classical swine fever virus (CSFV) genome, was expressed in porcine kidney cells PK-15, natural host of CSFV. In purifying cytoplasmic extracts from these cells by means of different concentrations of salt, glycerol and detergent four fractions, namely crude supernatant (SC) and different purified supernatants (S1, S2 and S3) were obtained. Using Western blot analysis the NS5B protein was found in all these fractions, showing that it was soluble in both higher and lower concentrations of salt, glycerol and detergent. The NS5B protein present in the four different fractions exhibited RNA-dependent RNA polymerase (RdRp) activity, but it was unable to complete the whole process of RNA synthesis. Site-directed mutation analysis showed that Thy216 and Cyt228 were essential for RNA synthesis while Cyt219 was not, suggesting that CSFV RdRp was template-specific. We conclude that initiation of RNA synthesis by CSFV RdRp includes also template priming. PMID- 14524474 TI - A sensitive and specific detection of Human herpesvirus 8 by polymerase chain reaction and dot blot hybridization. AB - Polymerase chain reaction (PCR) is a powerful technique of detecting Human herpesvirus 8 (HHV-8), but has a limited sensitivity and specificity. A new assay of HHV-8 based on combination of PCR with dot blot hybridization (DBH) was developed and evaluated for its sensitivity and specificity. An HHV-8-specific primer pair, ORF26out was used for amplification of target DNA. When the PCR product was detected visually the limit of detection was 0.1 ng DNA isolated from HHV-8-infected BC-3 cells. For DBH, the DNA amplified with the primer pair ORF26in specific for HHV-8 was labeled with digoxigenin (DIG). This DIG-labeled probe was capable of detecting 1.0 ng of DNA isolated from HHV-8-infected BC-3 cells. On the other hand, PCR combined with DBH (PCR/DBH) was more sensitive than PCR or DBH alone and also very specific. The sensitivity of PCR/DBH was higher than that of PCR and DBH alone. The PCR/DBH assay can be applied efficiently to confirm the presence of HHV-8 in clinical samples and to differentiate specifically HHV-8 infection from other viral infections. PMID- 14524475 TI - Lytic replication of Human herpesvirus 8 and induction of apoptosis. AB - Although many viruses can induce apoptosis in infected cells, large DNA viruses, such as poxviruses, herpesviruses and adenoviruses, usually exhibit the ability to suppress the induction of apoptosis in the infected cells. We investigated the ability of Human herpesvirus 8 (HHV-8) to protect cells from apoptosis induced by the virus. HHV-8 has been shown to harbor genes with anti-apoptotic capacity. However, we demonstrate here that a lytic replication of HHV-8 resulted in induction of apoptosis using different techniques to detect apoptosis. Therefore, despite the presence of anti-apoptotic genes in its genome, HHV-8 could complete its cycle of productive infection while inducing apoptosis in infected cells. This finding might have implications for the pathobiology of HHV-8 and other gamma herpesviruses in vivo. PMID- 14524476 TI - Sequence and phylogenetic analysis of SH, G, and F genes and proteins of Human respiratory syncytial virus isolates from Singapore. AB - To study the genetic variability and molecular epidemiology of Human respiratory syncytial virus (HRSV) occurring in Singapore, nucleotide sequencing of three membrane-associated genes (SH, G and F) of four local isolates was performed. Comparison of their nucleotide and amino acid sequences with those of the prototype strains A2 (subgroup A) and CH-18537 (subgroup B) indicated that the Singapore isolates belong to the subgroup A. Comparison of the Singapore isolates with the reference strain A2 showed that whereas the G protein was the most divergent with up to 15% difference, the F and SH proteins showed less diversity of only up to 4%. Each gene exhibited its distinct variable and conserved regions. The N- and O-glycosylation sites within the G protein of the isolates were analyzed to ascertain their potential implications on the antigenicity of the viral glycoprotein. Based on the second variable region of the G protein, phylogenetic analysis of the Singapore isolates with 91 previously identified genotypes of subgroup A revealed that more than one genotype (GA2 and GA5) may circulate in the local population at a given time. This epidemiological study reflects the pattern of genetic relationships between the HRSV isolates from Singapore to those from other parts of the world. PMID- 14524478 TI - Effect of monensin on Mayaro virus replication in monkey kidney and Aedes albopictus cells. AB - The effect of a cationic ionophore, monensin, on the replication of Mayaro virus in monkey kidney TC7 and Aedes albopictus cells has been studied. Treatment of these cells with 1 micromol/l monensin during infection did not affect the virus protein synthesis but inhibited severely the virus replication. Electron microscopy of the cells infected with Mayaro virus and treated with monensin revealed that the morphogenesis of Mayaro virus was impaired in TC7 but not in A. albopictus cells. PMID- 14524477 TI - Molecular characterization of a carnation etched ring virus isolate from India. AB - Incidence of the Carnation etched ring virus (CERV), the only DNA virus reported to date on carnation, was investigated by a bioassay using a partially purified virus as inoculum and then by a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Out of 61 carnation cultivars analyzed 41 (67%) were found positive. The virus positivity was verified by polymerase chain reaction (PCR) and nucleotide sequencing. The amplified 1349 bp fragment was by about 98% and 96% identical with respect to coat protein (CP) and enzymatic polyprotein genes, respectively, as compared to the sequences available in the database. In terms of amino acid sequence similarity, the homology values were 99% and 97%, respectively. Comparison with other caulimoviruses revealed that CERV is most closely related to the Cauliflower mosaic virus (CaMV). High genetic stability of CERV may be attributed to the fact that it has evolved from the same initial sequence in an original host. Because of global market of cut flowers and vegetative propagation it has been dispersed around the world. PMID- 14524479 TI - Cd38 expression on Cd8+ T cells in Human immunodeficiency virus 1-positive adults treated with HAART. AB - The aim of this study was to assess whether the density of CD38 antigen expression on CD8+ T cells can be used as a marker of activation of the immune system in Human immunodeficiency virus 1 (HIV-1)-positive patients treated with highly active antiretroviral therapy (HAART). T cell subsets, expression of CD38 antigen on CD8+T cells, HIV-1 viral load and stage of the disease were analyzed at baseline and after 12 months of HAART in 24 HIV-1-infected patients. Our data showed that the use of HAART is effective in reducing plasma viral load and in achieving a stable CD4+ count and percentage of CD8+/CD38+ cells. The percentages of CD8/CD38+ cells in HIV-1-infected patients at baseline and after 12 months of HAART were significantly higher than those of controls. Analysis of the density of CD38 expression revealed that it was due to CD8+/CD38+ subsets with low and medium density of antigen expression. Absolute number of CD4+ T cells correlated negatively with the percentage of CD8+/CD38+ cells at baseline of the study. Persistent up-regulation of the CD38 expression on CD8+ T cells and its correlation with the decreased CD4+ count despite the reduction of plasma viral load may reflect residual replication of HIV-1 in reservoirs. Thus, this immunological parameter can serve as a biological marker of HIV-1 infection and might have utility in clinical management of HIV-1-infected persons. PMID- 14524480 TI - Genotyping of Chikungunya virus isolates from India during 1963-2000 by reverse transcription-polymerase chain reaction. PMID- 14524481 TI - Further structural characteristics of the lipopolysaccharide from Coxiella burnetii strain nine mile in low virulent phase II. PMID- 14524482 TI - Permanent prostate brachytherapy in men with clinically localised prostate cancer. AB - Permanent prostate brachytherapy techniques are associated with excellent biochemical control for patients with localised prostate cancer. Ten-year data show that permanent prostate brachytherapy is compatible with external beam irradiation or radical prostatectomy. However, treatment protocols and techniques for prostate brachytherapy vary between centres and there is little conformity of treatment protocols. The selection of patients for monotherapy or combined external beam irradiation and brachytherapy is controversial. The role of neoadjuvant androgen deprivation also remains unanswered in patients with localised prostate cancer. In addition, post-implant dosimetry may in fact be more significant for predicting outcome than the addition of adjuvant therapies, and should be a requirement when performing prostate brachytherapy. Data now seem to support specific computed tomography (CT)-based criteria to evaluate implant quality and delivered dose to the prostate. Unfortunately, prostate oedema and poor imaging techniques are limiting factors for evaluating implant dosimetry. Treatment planning techniques that use new treatment planning computers may assist in improving the implant procedure and dosimetry and are now available. PMID- 14524483 TI - The role of hormone therapy as a neoadjuvant to radical prostate radiotherapy. PMID- 14524484 TI - Is bicalutamide equivalent to goserelin for prostate volume reduction before radiation therapy? A prospective, observational study. AB - INTRODUCTION: We compare the cytoreductive efficacy of bicalutamide or goserelin with no hormonal manipulation in prostate cancer before brachytherapy. MATERIALS AND METHODS: Transrectal ultrasound volume estimations were performed in clinic and during the brachytherapy-planning scan. Between volume estimations, patients received no hormonal treatment, bicalutamide 150 mg once daily or goserelin 3.6 mg every 28 days. RESULTS: Patients receiving no hormonal manipulation had a volume increase of 8% compared with an 8% volume reduction in the bicalutamide group and a 26% reduction in the goserelin group. As initial prostate volume was not equivalent in the three groups, a subgroup analysis was performed on patients who received active treatment for more than 3 months who had initial prostate volume less than 55 cm3. In this subgroup, a mean fall in prostate volume of 7%, occurred in the bicalutamide group compared with 21% in the goserelin group. In both the original and subgroup analysis, the cytoreductive efficacy of goserelin was significantly greater than bicalutamide (P < 0.0001). CONCLUSION: In the absence of data from randomised trials, comparing the efficacy of these agents, luteinising hormone-releasing hormone (LHRH) analogues remain the gold standard for cytoreduction before prostate brachytherapy. If the neoadjuvant efficacy of hormonal manipulation in external beam radiotherapy is dependent on prostate volume reduction, then LHRH analogues may also prove more effective in this neoadjuvant role. PMID- 14524485 TI - A randomised trial of cranberry versus apple juice in the management of urinary symptoms during external beam radiation therapy for prostate cancer. AB - AIMS: The aim of the study was to assess whether the oral intake of cranberry juice cocktail compared with apple juice was associated with a significant difference in urinary symptoms experienced during radical external beam radiation therapy (EBRT) for prostate carcinoma. MATERIALS AND METHODS: One hundred and twelve men with prostate cancer were randomised to either 354 ml cranberry juice or apple juice a day. Stratification was based on a history of a previous transurethral resection of prostate (TURP yes/no) and baseline International Prostate Symptom Score (IPSS < 6 or > or = 6) of urinary symptoms. RESULTS: The maximum IPSS (MRT) and the maximum change in IPSS from baseline (DRT) are used to report the results. We analysed the effects of juice allocation on DRT and MRT using analysis of covariates (ANCOVA). We observed no significant difference for DRT (P = 0.39) or MRT (P = 0.76) related to the consumption of cranberry compared with apple juice. However, we found a significant relationship between the history of a previous TURP and both DRT (P = 0.01) and MRT (P = 0.01). The history of a previous TURP was associated with lower values for both end points. Baseline IPSS was significant for DRT (P = 0.004) and MRT (P < or = 0.001). We found a significant relationship between the baseline IPSS < 6 or > or = 6 cut point on MRT (P < or = 0.001) but not on DRT (P = 0.43). The use of neoadjuvant hormones had no significant effect on DRT (P = 0.64) or MRT (P = 0.76). The use of additional symptomatic medication during the study was not significantly different between the two arms. CONCLUSIONS: This study shows no significant difference in the urinary symptoms experienced during EBRT related to the consumption of cranberry juice compared with apple juice. PMID- 14524486 TI - Changes in clinical presentation, management and outcome in 1348 patients with differentiated thyroid carcinoma: experience in a single institute in Hong Kong, 1960-2000. AB - The clinical features, management and outcome of 1348 patients diagnosed with differentiated thyroid carcinoma in Queen Elizabeth Hospital, Hong Kong, were analysed according to the period of diagnosis: A (before 1980), B (1981-1990) and C (1991-2000). As time advanced, ratio of papillary carcinoma (PTC) to follicular carcinoma (FTC) increased (A:B:C = 1.6: 3.1: 7.2). The mean size of the primary tumour decreased (A:B:C = 3.5 cm: 2.8 cm: 2.5 cm), with a greater percentage of microcarcinoma of 1 cm or less (A:B:C = 5.1%: 16.1%: 21.7%). At presentation, the incidence of lymph-node metastasis decreased (A:B:C = 32.7%: 31.6%: 24.8%) and that of distant metastasis decreased (A:B:C = 9%: 6.1%: 5.3%). Bilateral surgical resection was more commonly used (A:B:C = 62.8%: 89.1%: 94.8%) than lobectomy (A:B:C = 26.3%: 2.8%: 1.8%). Radiation treatment, radioactive iodine (131I; RAI) and external radiotherapy (EXT), was more commonly used (A:B:C = 53.2%: 74.7%: 85.1%). RAI was used in 84.3% (A:B:C = 50%: 71.2%: 84.3%) and EXT in 14.5% of patients in the past decade (A:B:C = 10.9%: 8.7%: 14.5%). The proportion of patients who adopted a bilateral surgery and RAI treatment increased gradually with time (A:B:C = 33%: 68%: 83.8%). The 5-year cause-specific survival (A:B:C = 90.2%: 93.7%: 95.7%), locoregional failure-free survival (A:B:C = 72.6%: 82.9%: 91.6%) and distant metastasis failure-free survival (A:B:C = 84.5%: 89.1%: 92.6%) were improved. However, the period of diagnosis was not found to be an important explanatory variable (i.e. P > 0.05) in Cox regression after adjusting for other factors, indicating that the improvement was probably related to the temporal trend of other factors: presentation at earlier stage, increased ratio of PTC:FTC and more aggressive management by bilateral surgery and radiation therapy. PMID- 14524487 TI - External beam radiotherapy in the management of differentiated thyroid cancer. AB - AIMS: No randomised trials have addressed the use of external beam radiotherapy (EBRT) in the treatment of differentiated thyroid cancer. The indications for EBRT, the technique and recommended dose all remain controversial. MATERIALS AND METHODS: We included patients treated with EBRT with curative intent from two cancer centres between 1988 and 2001. Data were collected from hospital notes, radiotherapy prescriptions and local cancer registry. RESULTS: The indications for treatment in the 41 identified patients were macroscopic residual disease 23 (56%), microscopic residual disease 10 (25%), Hurthle cell variants 3 (7%), multiple lymph-node involvement 3 (7%) and other 2 (5%). Delivered doses ranged from 37.5-66 Gy over 3-6.5 weeks. Rate of local recurrence and overall survival at 5 years were as follows: papillary 26% and 67%; follicular 43% and 48%; well differentiated 21% and 67%; focus of poor differentiation/Hurthle cell variants 69% and 32%; complete excision 25% and 61%; residual disease 37% and 59%; EBRT total dose < 50 Gy 63% and 42%; 50-54 Gy 15% and 72%; > 54 Gy 18%, and 68%. CONCLUSIONS: The results in this study are consistent with previous retrospective studies of EBRT. The wide range of indications and doses used with radical intent highlights the lack of clinical and radiobiological data on the response of differentiated thyroid cancer to EBRT. Despite the small study size, the 5-year local recurrence results indicate a possible dose response within the dose range used. PMID- 14524488 TI - Radiotherapy for painful bone metastases: an overview of the overviews. PMID- 14524490 TI - Capacity and demand models for radiotherapy treatment machines. AB - Models to predict the number of linear accelerators (linacs) required usually assume that capacity needs to equal demand. Queuing theory shows that capacity needs to exceed mean demand, to avoid the build-up of waiting times. A model has been developed, using Monte-Carlo modelling, to calculate the percentage of spare capacity required to keep waiting times to treatment short. For a matched pair of linacs, in a department that closes on bank holidays and compensates for category 1 patients by treating twice before or after the break, about 10% spare capacity is required to ensure that 86% of patients are able to start radiotherapy within a week of completing the treatment planning process. If a machine is booked as a single (unmatched machine), an additional 3% spare capacity is needed. If all bank holidays in the year are worked, then about 3% less is needed. PMID- 14524489 TI - Palliation of metastatic bone pain: single fraction versus multifraction radiotherapy--a systematic review of randomised trials. AB - Recent randomised studies have reported that single fraction radiotherapy is as effective as multifraction radiotherapy in relieving pain caused by bone metastasis. However, there are concerns about the higher re-treatment rates and the efficacy of preventing future complications, such as pathological fracture and spinal cord compression, by single fraction radiotherapy. A systematic review of randomised studies, examining the effectiveness of single fraction radiotherapy versus multiple fraction radiotherapy for metastatic bone pain relief and prevention of bone complications, was conducted to help answer this controversy. Randomised studies comparing single fraction radiotherapy with multifraction radiotherapy on metastatic bone pain were identified. The analyses were performed using intention-to-treat principle. The results were pooled using meta-analysis to estimate the effect of treatment on pain response, re-treatment rate, pathological fracture rate and spinal cord compression rate. Twelve trials involving 3621 sites were included in the meta-analysis. The overall pain response rates for single fraction radiotherapy and multifraction radiotherapy were 60% (1080/1814) and 59% (1060/1807), respectively, giving an odds ratio (OR) of 1.03 (95% confidence interval [CI] 0.90-1.19), indicating no difference between the two radiotherapy schedules. There was also no difference in complete pain response rates for single fraction radiotherapy (34% [508/1476]) and multifraction radiotherapy (32% [475/1473]), with an OR of 1.10 (950% CI 0.94 1.30). Patients treated by single fraction radiotherapy had a higher re-treatment rate, with 21.5% (267/1240) requiring re-treatment compared with 7.4% (91/1236) of patients in the multifraction radiotherapy arm (OR 3.44 [95% CI 2.67-4.43]). The pathological fracture rate was also higher in single fraction radiotherapy arm patients. Three per cent (37/1240) of patients treated by single fraction radiotherapy developed pathological fracture compared with 1.6% (20/1236) for those treated by multifraction radiotherapy (OR 1.82 [95% CI 1.06-3.11]). The spinal cord compression rates were similar for both arms (OR 1.41 [95% CI 0.72 2.75]). Single fraction radiotherapy was as effective as multifraction radiotherapy in relieving metastatic bone pain. However, the re-treatment rate and pathological fracture rate were higher after single fraction radiotherapy. Studies with quality of life and health economic end points are warranted to find out the optimal treatment option. PMID- 14524491 TI - A method of predicting workload and staffing level for radiotherapy treatment planning as plan complexity changes. AB - When assessing the impact on workload from an expanding number of new patients and increasing treatment complexity, radiotherapy clinics find that oversimplified workload parameters, such as number of patients or number of fields, are not suitable for managing and predicting workload and organisational dimension. Although the basic treatment equivalent concept is available for predicting linear accelerator utilisation, no corresponding parameter has been available for radiotherapy treatment planning. In this study, we derive a simple workload indicator for treatment planning. The dose plan unit (DPU) takes the complexity of the treatment plan into account. Categorising plans according to complexity in their production, and measuring corresponding time for completing the plans in these categories, leads to the following baseline values for workload prediction: 1 DPU (non-computed tomography [CT]), 3 DPU (CT-contour) and 6 DPU (full-CT). The measured average productivity of 0.65 DPU per hour (1 standard deviation, SD = 0.08 DPU), or alternatively 1 DPU = 92 min, for a dosimetrist in this clinic indicates that 79.0 DPU can be produced by each dosimetrist per month within normal working hours. The predictive power of the DPU is shown in terms of using it to quantify the impact on workload in treatment planning of changing treatment protocols for a particular anatomic treatment site. PMID- 14524492 TI - Primary chondroid chordoma of the petrous part of the temporal bone. PMID- 14524493 TI - Fatal hepatic veno-occlusive disease in an adult patient with Wilms' tumour. PMID- 14524494 TI - Pasteurella multocida infection following cisplatin and 5-fluorouracil chemotherapy. PMID- 14524495 TI - AT heterozygosity in a patient with multiple myeloma. PMID- 14524496 TI - Detection of gill-associated virus (GAV) by in situ hybridization during acute and chronic infections of Penaeus monodon and P. esculentus. AB - Chronic and acute gill-associated virus (GAV) infections were examined by in situ hybridization (ISH) using a DNA probe targeting a 779 nucleotide region of the ORF1b-gene. Chronic GAV infections were observed in healthy Penaeus monodon collected from farms and healthy P. esculentus surviving experimental infection. During chronic-phase infections in both species, GAV was detected only in partitioned foci of cells with hypertrophied nuclei (spheroids) within the lymphoid organ. Acute-phase infections were observed in moribund P. monodon and P. esculentus infected experimentally with a high dose of GAV, and in moribund P. monodon collected from farms during outbreaks of disease. During acute experimental infections in P. monodon, ISH detected GAV throughout the lymphoid organ, in gills and in connective tissues throughout the cephalothorax. In moribund P. monodon collected from natural outbreaks of disease, GAV was also detected in the gills and in connective tissues of the cephalothorax, but the distribution of virus within the lymphoid organ varied. In acutely infected P. esculentus, GAV was detected in connective tissues, but was restricted to the inner stromal matrix cells and endothelial cells of intact lymphoid organ tubules. The tissue distribution of GAV identified by ISH suggests that shrimp are able to control and maintain chronic asymptomatic infection by a process involving lymphoid organ spheroids. Acute phase infections and the development of disease appear to be dose-related and involve the systemic distribution of virus in connective tissues throughout the cephalothorax. PMID- 14524497 TI - Emergence and maintenance of infectious salmon anaemia virus (ISAV) in Europe: a new hypothesis. AB - The present study describes the use of molecular methods in studying infectious salmon anaemia virus (ISAV), an important pathogen of farmed salmon in Norway, Scotland, the Faeroe Islands, Canada, USA and Chile. The nucleotide sequences of the haemagglutinin gene (HA) from 70 ISAV isolates have been analysed for phylogenetic relationship and the average mutation rate of nucleotide substitutions calculated. The isolates constitute 2 major groups, 1 European and 1 North American group. The isolate from Chile is closely related to the North American isolates. The European isolates can be further divided into 3 separate groups reflecting geographical distribution, time of collection, and transmission connected with farming activity. Based on existing information about infectious salmon anaemia (ISA) and new information emerging from the present study, it is hypothesised that: (1) ISAV is maintained in wild populations of trout and salmon in Europe; (2) it is transmitted between wild hosts mainly during their freshwater spawning phase in rivers; (3) wild salmonids, mainly trout, possibly carry benign wild-type ISAV isolates; (4) a change (mutation) in virulence probably results from deletions of amino acid segments from the highly polymorphic region (HPR) of benign wild-type isolates; (5) ISA emerges in farmed Atlantic salmon when mutated isolates are transmitted from wild salmonids or, following mutation of benign isolates, in farmed salmon after transmission from wild salmonids; (6) farming activity is an important factor in transmission of ISAV between farming sites in addition to transmission of ISAV from wild salmonids to farmed salmon; (7) transmission of ISAV from farmed to wild salmonids probably occurs less frequently than transmission from wild to farmed fish due to lower frequency of susceptible wild individuals; (8) the frequency of new outbreaks of ISA in farmed salmon probably reflects natural variation in the prevalence of ISAV in wild populations of salmonids. PMID- 14524498 TI - Experimental vertical transmission of Piscirickettsia salmonis and in vitro study of attachment and mode of entrance into the fish ovum. AB - Piscirickettsia salmonis is a pathogenic bacterial agent causing septicaemic disease in salmon. Since its isolation in Chile in 1989, P. salmonis has continually produced high mortality rates in salmon farms. Little information exists regarding the mechanisms of vertical transmission of this pathogen. Experimental vertical transmission was established in the present study by inoculation of male and female rainbow trout broodstock with P. salmonis. The bacterium was subsequently detected using indirect immunofluorescence in milt and coelomic fluid of the majority of inoculated broodstock (14/15). Bacteria were detected in the fry when 1 or both parents were inoculated, although none of the infected fry presented signs of the disease. P. salmonis was also detected in progeny obtained through fertilisation ova from non-inoculated females incubated in a medium containing a bacterial suspension, demonstrating transmission during the process of fertilisation. Ova infected in vitro were examined at sample periods from 30 s to 60 min using scanning electron microscopy. This demonstrated that the bacterium attaches to the ova by means of membrane extensions, structures which we have called 'piscirickettsial attachment complex' (PAC) and which would allow later penetration into the ovum. PMID- 14524499 TI - Vaccine efficacy in spotted wolffish Anarhichas minor: relationship to molecular variation in A-layer protein of atypical Aeromonas salmonicida. AB - Atypical Aeromonas salmonicida strains comprise a heterogeneous group in terms of molecular and phenotypic characteristics. They cause various conditions of ulcer diseases or atypical furunculosis and are being isolated in increasing number from various fish species and geographical areas. Several marine fish species susceptible to atypical A. salmonicida, including spotted wolffish Anarhichas minor O., are now being farmed and new vaccines may be needed. A commercial furunculosis vaccine for salmon is reported to protect wolffish poorly against experimental challenge with atypical A. salmonicida. The protective antigen(s) in furunculosis vaccines is still unclear, but in oil-adjuvanted vaccine for Atlantic salmon Salmo salar L., the surface A-layer was shown to be important for protection. In spotted wolffish, the efficacy of atypical furunculosis vaccines seems to vary with the atypical A. salmonicida strains used as bacterin in the vaccine. In the present study we investigated whether differences in the A-layer protein among atypical strains might be responsible for the observed variation in vaccine efficacy. Atypical A. salmonicida strains from 16 fish species in 11 countries were compared by genome polymorphism analysis using amplified fragment length polymorphism (AFLP) fingerprinting and by comparative sequencing of the vapA genes encoding the A-protein. The A-protein sequences appeared to be highly conserved except for a variable region between Residues 90 to 170. Surprisingly, the grouping of strains based on AFLP- or A-protein sequence similarities was consistent. In addition, serological differences in the A-protein among the strains were demonstrated by an A-protein-specific monoclonal antibody. Vaccines based on atypical A. salmonicida strains possessing genetically and serologically different A-layer proteins were shown to result in significantly different protection in spotted wolffish. PMID- 14524500 TI - CpG motif in synthetic ODN primes respiratory burst of olive flounder Paralichthys olivaceus phagocytes and enhances protection against Edwardsiella tarda. AB - Effects of synthetic cytidine-phosphate-guanosine (CpG) oligodeoxynucleotide (ODN) on respiratory burst activity of olive flounder (Paralichthys olivaceus) head-kidney phagocytes and on protection against lethal infection with Edwardsiella tarda were investigated. Phagocytes precultured with a CpG ODN showed significantly higher chemiluminescence (CL) responses than phagocytes precultured with guanosine-phosphate-cytidine (GpC) ODN or culture medium alone (control) at all concentrations. Supernatants produced from leucocytes, which were pulsed with CpG ODN, induced significantly higher respiratory burst activity than supernatants produced by GpC ODN or culture medium alone. In an in vivo experiment, respiratory burst activities of the head kidney phagocytes in the groups injected either 0.25 or 0.5 microg fish(-1) of CpG ODN were significantly higher than those in the groups injected with GpC ODN or HBSS (control) at 3, 5 and 7 d after injection. The groups of fish injected with 0.25 or 0.5 microg of CpG ODN showed higher survival rates (83.3%) than groups treated with GpC ODN (33.3%) and a control group (8.3%) after challenge with E. tarda. The present in vitro and in vivo experiments have demonstrated the ability of synthetic CpG ODN to increase phagocyte respiratory burst activity and disease resistance in olive flounder. PMID- 14524501 TI - In vitro nutritional requirements and metabolic products of pathogenic and nonpathogenic strains of Cryptobia salmositica: essential carbohydrates and amino acids. AB - Pathogenic and nonpathogenic strains of Cryptobia salmositica cultured in minimum essential medium (MEM) with several monosaccharides, disaccharides and amino acids were observed for differences in multiplication and motility. Metabolic end products (i.e. alanine, aspartate, carbon dioxide, lactate and pyruvate) were measured for logarithmically growing cells under aerobic conditions. The pathogenic strain of C. salmositica multiplied more readily in MEM supplemented with D(-)ribose, D(+)xylose, D(+)galactose, D(+)glucose, D(+)mannose and D( )fructose. However, there were no significant differences in multiplication when the strains were cultured with the monosaccharide D(-)arabinose. The nonpathogenic strain multiplied significantly better than the pathogenic strain in the presence of the disaccharides alpha-lactose, maltose and sucrose. It also multiplied more readily when the amino acids L-glutamine and D(-)proline were added to MEM. The end products of carbohydrate catabolism under aerobic conditions were alanine, aspartate, carbon dioxide, lactate and pyruvate. PMID- 14524502 TI - Cryo-archiving of Batrachochytrium dendrobatidis and other chytridiomycetes. AB - Batrachochytrium dendrobatidis is a major pathogen of frogs worldwide. It has been associated with catastrophic declines of frog populations including those in pristine habitats in Queensland, Australia. To facilitate genetic and disease studies of this fungus and related species, it is essential to have a reliable long-term storage method to maintain genetic integrity of isolates. We have adapted well-established techniques used for the long-term storage of tissue culture cell lines to the preservation of B. dendrobatidis and other chytridiomycetes. This simple method has allowed us to recover these fungi from storage at -80 degrees C and in liquid nitrogen over an extended period. With this technique it is now possible to preserve saprobic and parasitic isolates from a variety of environmental and disease situations for comparative genetic and biological studies. PMID- 14524503 TI - Parasitism by Dermocystidium ranae in a population of Rana esculenta complex in Central Italy and description of Amphibiocystidium n. gen. AB - We report the enigmatic parasite Dermocystidium ranae in a green frog population (Solomeo, Umbria, Italy) of the Rana esculenta complex, consisting of the parental species R. lessonae (L) and hybrid form R. esculenta (E). In this population a rapid 50% decline of the parental form L was observed. Large dermal U-shaped cysts of D. ranae were found primarily on the ventral aspect of infected individuals, with a significantly higher incidence of infection in the parental species compared to the clonal hybrid. In each form, however, there was little pathological change associated with infection, and the cause of the recent declines of R. lessonae at this site remains unknown. In this paper we present the first ultrastructural description of an amphibian Dermocystidium sp. and we review the taxonomy of Dermocystidium, Dermosporidium and Dermomycoides spp. from amphibians. We conclude that Dermosporidium multigranulare Broz & Kulda, 1954 is synonymous with Dermocystidium ranae Guyenot & Naville, 1922 and, due to lack of sufficient differences between genera and significant dissimilarities with fish Dermocystidium spp., the 3 amphibian genera are synonymous. We propose that they should be designated to a new genus, Amphibiocystidium n. gen., and Dermocystidium retained for those species parasitic in fish. PMID- 14524504 TI - Systematic evaluation of factors controlling Perkinsus marinus transmission dynamics in lower Chesapeake Bay. AB - The transmission of Perkinsus marinus in eastern oysters Crassostrea virginica in relation to water temperature, host oyster mortality, and water-column abundance of anti-P. marinus antibody-labeled cells was systematically examined for 20 mo at a site in the lower York River, Virginia, USA. Uninfected sentinel oysters were naturally exposed to the parasite at 2 wk intervals throughout the course of the study to determine the periodicity and rates of parasite transmission. The timing and magnitude of disease-associated oyster mortalities in a local P. marinus-infected oyster population were estimated by monitoring a captive subset of the local oyster population. Flow cytometric immunodetection methods were employed to estimate the abundance of P. marinus cells in water samples collected 3 times each week. The acquisition of P. marinus infections by naive sentinel oysters occurred sporadically at all times of the year; however, the highest incidence of infection occurred during the months of August and September. This window of maximum parasite transmission coincided with the death of infected hosts within the captive local oyster population. Counts of antibody-labeled cells ranged from 10 to 11900 cells l(-1), with the highest abundances in July and August coincident with maximum summer temperatures. A statistically significant relationship between water-column parasite abundance and infection acquisition rate was not observed; however, highest parasite-transmission rates in both years occurred during periods of elevated water-column abundance of parasite cells. These results support the prevailing model of P. marinus transmission dynamics by which maximum transmission rates are observed during periods of maximum P. marinus-associated host mortality. However, our results also indicate that transmission can occur when host mortality is low or absent, so alternative mortality-independent dissemination mechanisms are likely. The results also suggest that atypically early-summer oyster mortality from Haplosporidium nelsoni infection, at a time when infections of P. marinus are light, has a significant indirect influence on P. marinus transmission dynamics. Elimination of these hosts prior to late-summer P. marinus infection intensification effectively reduces the overall number of P. marinus cells disseminated. PMID- 14524505 TI - First description of cardiomyopathy syndrome (CMS)-related lesions in wild Atlantic salmon Salmo salar in Norway. AB - This report represents the first diagnosis of cardiomyopathy syndrome (CMS) in migrating, wild Atlantic salmon Salmo salar from 1 major river and off the coast of Norway. Previously, this disease has been diagnosed only in farmed Atlantic salmon. The possible significance of the disease in wild stocks of salmon is discussed. PMID- 14524506 TI - Effects of cadmium and zinc toxicity on orientation behaviour of Echinoparyphium recurvatum (Digenea: Echinostomatidae) cercariae. AB - The effects of cadmium and zinc toxicity on orientation behaviour (photo- and geo taxis) of Echinoparyphium recurvatum cercariae was investigated at concentrations ranging from 10 to 1000 microg l(-1). Exposure to the toxicants at all metal concentrations caused a change in orientation to negative phototaxis and positive geotaxis during the submaximal dispersal phase (0.5 h cercarial age). Autometallography staining of cercariae exposed to 1000 microg l(-1) cadmium or zinc showed selective binding of heavy metals to tegumental surface sites associated with sensory receptors. The significance to parasite transmission of changes in cercarial orientation behaviour in metal polluted environments is discussed. PMID- 14524507 TI - Reliability of the clinical examination: how close is "close enough"? PMID- 14524508 TI - A modified neuromuscular electrical stimulation protocol for quadriceps strength training following anterior cruciate ligament reconstruction. AB - STUDY DESIGN: Randomized clinical trial, single-masked. OBJECTIVES: To determine the effectiveness of using a modified neuromuscular electrical stimulation (NMES) training program as an adjunct treatment for improving quadriceps strength and physical function in rehabilitation following anterior cruciate ligament reconstruction (ACLR). BACKGROUND: NMES training for quadriceps strengthening has previously been shown to be an effective adjunct treatment following ACLR when performed against isometric resistance using a dynamometer with the knee positioned in flexion. We developed a modified version of published NMES protocol because some patients have difficulty tolerating the existing protocol and many clinics may not have instrumented dynamometers. There is a need to determine the effectiveness of this modified protocol. METHODS AND MEASURES: Forty-three subjects who had undergone ACLR were randomly assigned to either a group that received (NMES group) or did not receive (comparison group) the NMES treatment in conjunction with their rehabilitation. Group means for quadriceps strength and self-reported measures of knee function were compared after 12 and 16 weeks of rehabilitation. The proportion of subjects in each group achieving clinical criteria to initiate ambulation without crutches, treadmill running, and agility training at selected times during rehabilitation were also compared. RESULTS: The NMES group demonstrated moderately greater quadriceps strength at 12 weeks (effect size, 0.48), and moderately higher levels of self-reported knee function at both 12 (effect size, 0.72) and 16 (effect size, 0.65) weeks of rehabilitation compared to the comparison group. A greater proportion of subjects in the NMES group achieved clinical criteria for advancing to agility training at 16 weeks. CONCLUSIONS: The modified NMES quadriceps training protocol can be a useful adjunct to ACLR rehabilitation programs, but the treatment effect is smaller than what has been reported in previous studies. PMID- 14524509 TI - Muscle energy technique in patients with acute low back pain: a pilot clinical trial. AB - STUDY DESIGN: A prospective, pilot clinical trial. OBJECTIVE: Examining the outcomes of Muscle Energy Technique (MET) in patients with acute low back pain. BACKGROUND: MET is commonly used to treat patients with acute low back pain. No randomized controlled trials examining the outcomes of this treatment in symptomatic populations has been reported in the literature. METHODS AND MEASURES: Ten men and 9 women diagnosed with acute low back pain were randomly assigned with stratification to 1 of 2 treatment groups. Patients were matched according to age, gender, and initial Oswestry score. The control group received supervised neuromuscular re-education and resistance training while the experimental group received the same exercises coupled with MET. Both groups received the selected treatment 8 times over a 4-week period (2 times per week). Patients completed an Oswestry Disability Index on their first and eighth visits and change scores were calculated. RESULTS: A 2-tailed t test (P < .05) demonstrated a statistically significant difference with the experimental group showing greater improvement in the Oswestry Disability Index score than the control group. CONCLUSION: MET combined with supervised motor control and resistance exercises may be superior to neuromuscular re-education and resistance training for decreasing disability and improving function in patients with acute low back pain. PMID- 14524510 TI - Children and adolescents with low back pain: a descriptive study of physical examination and outcome measurement. AB - STUDY DESIGN: A retrospective, cohort study of children and adolescents with low back pain (LBP) referred to physical therapy. OBJECTIVES: To describe the clinical presentation, examination findings, and classification of children and adolescent patients with LBP, and to explore the concurrent validity of the Oswestry questionnaire as a disability measure in this group. BACKGROUND: Little information is available regarding the clinical presentation, physical examination findings, and outcome assessment in children and adolescents with LOP. METHODS AND MEASURES: Charts were reviewed for historical, diagnostic, physical examination,and outcome information. Diagnoses given by the referring physicians were recorded. A treatment-based classification was made for each subject. Descriptive statistics were calculated for all variables. The validity of the Oswestry questionnaire was examined. RESULTS: The children and adolescents included in this study represented 5% (n = 25) of all cases of LOP referred to physical therapy. A greater percentage of patients had difficulty with extension than with flexion range of motion (ROM). Initial pain scores were lower if a specific pathology was present (P = .001). Initial pain and Oswestry scores were poorly correlated (r = 0.16). Forty-four percent (n = 11) of patients scored under the floor value of 12% on the Oswestry. CONCLUSION: The referral rate of children and adolescents with LBP seems to be low. Compared to adults, children and adolescents appear more likely to have a specific diagnosis given to them by their physician. The physical examination findings appear to indicate that spinal stabilization approaches may be beneficial for many patients. The Oswestry questionnaire may not be a valid tool for documenting changes in disability in these patients. Further research is needed on the conservative management of children and adolescents with LBP. PMID- 14524511 TI - Reliability of 2 functional goniometric methods for measuring forearm pronation and supination active range of motion. AB - STUDY DESIGN: Test-retest reliability study. OBJECTIVES: To determine intra- and intertester reliability of the hand-held pencil (HHP) and the plumbline goniometer (PLG) methods for measuring active forearm pronation and supination motions in individuals with and without injuries. BACKGROUND: The distal forearm method has been considered the gold standard for measuring forearm pronation and supination motion. The HHP and PLG, however, are 2 more functional methods for measuring forearm motions, though limited information on the psychometric properties of these tests is currently available. METHODS AND MEASURES: Intra- and intertester reliability of the HHP and PLG methods were determined in 40 subjects of convenience (20 injured and 20 noninjured). Two testers performed 3 repeated measurements for each motion and method on all subjects. Intraclass correlation coefficients (ICC3,1 for intratester reliability, ICC2,3 for intertester reliability) and standard error of measurements (SEMs) were determined. RESULTS: The ICCs for the measurements of pronation and supination using the HHP and PLG methods were high (range, 0.86-0.98) for individuals with and without injuries, with the reliability for the PLG method being equal or slightly greater than the HHP method for the majority of pronation and supination measurements. Intratester ICCs were higher (SEMs were conversely lower) than intertester ICCs for nearly all measurements. The ICC values were generally the same or higher for individuals with injuries compared to individuals without injuries. CONCLUSIONS: The HHP and PLG are highly reliable methods for measuring functional forearm pronation and supination. Because plumbline goniometers are not commercially available and the instrumentation for the HHP method is readily accessible, clinicians should consider the latter as their method of choice for measuring functional forearm pronation and supination. PMID- 14524512 TI - The effects of treadmill type on heart rate and pain threshold velocity in individuals with lower-extremity musculoskeletal pain. AB - STUDY DESIGN: This study utilized a quasi-experimental design in which subjects served as their own controls. OBJECTIVE: To determine whether heart rate, pain threshold velocity, and pain perception varied in patients running on a soft-belt treadmill versus a standard hard-belt treadmill. BACKGROUND: According to promotional literature, the relatively new Orbiter soft-belt tread produces a greater increase in heart rate at a given velocity as well as a higher velocity tolerance while walking or running. The manufacturer also asserts that decreased forces transmitted through the lower extremity should decrease pain levels while exercising on the soft-belt treadmill. METHODS AND MEASURES: Twenty-seven subjects walked or ran on each of 2 treadmills at incrementally increasing velocities until they experienced either the onset of pain or an increase in pain from baseline levels. Locomotion continued for 2 minutes after that, during which time heart rate and pain level on a visual analog scale (VAS) were recorded. RESULTS: Two univariate paired t tests and a Wilcoxon's signed rank test revealed a greater heart rate and pain threshold velocity when using the soft-belt treadmill with no statistical difference in the pain reported between the 2 treadmills. CONCLUSION: Our study revealed a 10% higher heart rate and a 14.5% higher pain threshold velocity with the soft-belt treadmill compared to a hard belt treadmill. These differences are considered clinically meaningful. PMID- 14524514 TI - Soft-tissue arthroplasty for hallux rigidus. AB - Seven patients (seven feet) were evaluated at an average follow-up of 42 months following soft-tissue interposition arthroplasty of the hallux metatarsophalangeal (MTP) joint for severe hallux rigidus. The technique involved reaming of the base of the proximal phalanx and metatarsal head to decompress the first MTP joint and placement of a soft-tissue tendon bundle as a biologic spacer. Six of seven patients had bilateral disease, and a positive family history of hallux rigidus. At final follow-up, all seven patients rated their result as good or excellent, the level of pain was substantially reduced, and the mean AOFAS score substantially improved from 46 to 86 points. Mild metatarsalgia was noted by four patients and characterized by mild plantar callosities in these cases. The mean MTP dorsiflexion improved from 9 degrees to 34 degrees and patients demonstrated good to excellent plantarflexion strength on manual muscle testing and with toe rise. Physical examination of the involved feet demonstrated no evidence of pes planus, metatarsus primus elevatus, Achilles tendon contracture, or metatarsocuneiform joint hypermobility in any of the seven. The technique of soft-tissue interposition arthroplasty as described gave excellent pain relief and reliable function of the hallux, and is an alternative treatment to MTP arthrodesis in select cases of severe hallux rigidus. PMID- 14524515 TI - Flexor hallucis longus transfer for repair of chronic achilles tendinopathy. AB - BACKGROUND: The flexor hallucis longus (FHL) tendon has been used to augment the repairs for chronic Achilles tendinopathy. Two common methods of FHL harvesting include a single incision (posterior) technique and a double incision (posterior and medial utility) technique. This cadaver study was designed to measure and compare the lengths of FHL tendon obtainable for reconstruction with each technique. METHODS: Fourteen fresh-frozen cadaver lower limbs were utilized for FHL harvest. The tendon was first exposed through the single posterior-medial incision approach adjacent to the Achilles. A second medial utility midfoot incision was then made and the FHL was marked at the level of Henry's knot with a suture, to approximate the level of potential harvest via a two-incision technique. The FHL was then harvested and delivered into the posterior wound. Single incision technique graft length was then measured from the tip of the calcaneal tuber to the level of transection. The remaining in situ tendon was then also measured between its level of transection and the more distal suture placed at Henry's knot. These two lengths were then combined to determine the total potential tendon graft length obtainable using a double incision technique. RESULTS: The average length of the FHL tendon harvested through the single posterior incision technique measured 5.16 cm (range, 3.4-6.9 cm, SD = 1.29). The average total tendon graft length available using the double incision technique measured 8.09 cm (range, 5.1-11.1 cm, SD = 1.63). The difference between the lengths obtained from these two techniques was significant (p < .001). CONCLUSIONS: These results demonstrate approximate FHL graft lengths obtainable by using either a single or double incision harvest technique and show that a significantly longer graft can be obtained using a double incision technique. Further data need to be obtained, however, to support whether the extra surgery and graft length obtained from a double incision technique are of any benefit in improving the ultimate functional outcome of these repairs. PMID- 14524516 TI - Ankle stabilization with hamstring autograft: a new technique using interference screws. AB - Many techniques have been advocated for the late surgical reconstruction of the lateral ankle ligament complex, including the use of hamstring tendon autografts. A new technique is described which attempts to restore anatomically the anterior talofibular ligament using hamstring autograft. The distal end of the graft is secured in a blind-ended tunnel in the talus without the need for transosseous tensioning. This is achieved with the use of the Bio-Tenodesis Screw System. PMID- 14524517 TI - Outcome and recurrence rate of diabetic foot ulcers treated by a total contact cast: short-term follow-up. AB - Foot ulcers in patients with diabetes present an increasing burden to healthcare. Total contact cast (TCC) is a well established treatment option for many of these ulcers, but little seems to be known about the final outcome once healing has been reached. The study examined healing and final outcome after TCC treatment among patients with diabetes. During a 22-month period, 15 consecutive patients with a total of 17 ulcers started treatment with a TCC. Three patients (three ulcers) were lost to follow-up (FU). Average FU was 91 weeks. The patients were followed with emphasis on healing, complications during FU, footwear, compliance, and frequency of visits. All ulcers healed, some after additional surgery. During FU, control visits were performed at an average of one visit every 4.7 weeks. Notwithstanding this, a high frequency of ulcer recurrence and/or other complications was noticed. Only four patients out of 12 remained without any new (ulcer) problem during FU. TCC proved to be a very efficient tool to heal neuropathic foot ulcers, but the recurrence rate and frequency of other complications remained very high. These data show how difficult it is to achieve reliable (secondary) preventive general foot care, and to prescribe and manufacture reliable footwear. PMID- 14524518 TI - Bilobed flaps for nonhealing ulcer treatment. AB - Healing of round ulcers may be difficult particularly in the plantar area. Rigidity and thickness of the plantar skin do not allow fusiform excision and primary suturing. The bilobed flap is a simple reconstructive technique principally used to repair substantial defects in the facial region. The authors' experience with this local flap in the foot is presented with good short-term results. Between 1995 and 1998, five female and seven male neuropathic foot patients with round plantar ulcers were treated with bilobed flaps. The average age of the patients was 50 (range, 15-76). The average size of the ulcers was 1.6 cm (1-3.2 cm). Debridement and orthotic insoles were used at least for 3 months before considering bilobed flaps. Seven patients were diagnosed as type II diabetes mellitus, four patients had cerebral palsy, and another patient had meningomyelocele. The minimal follow-up period was 1 year (average, 19.5 months). The only complication was wound dehiscence at the lateral side of the heel in a type II diabetic. Subsequently, this complicated ulcer was managed with a sliding flap and skin graft without further problem. The study concluded that nonhealing foot ulcers can be effectively treated with a bilobed skin flap of healthy tissues rotated from nonweightbearing parts of the sole. PMID- 14524519 TI - Minor environmental trauma and lower extremity amputation in high-risk patients with diabetes: incidence, pivotal events, etiology, and amputation level in a prospectively followed cohort. AB - This study determined the incidence, pivotal events, etiology, and levels of amputation in a prospectively followed cohort of 400 people with diabetes and a prior healed foot ulcer who participated in a randomized footwear trial. Participants were seen every 17 weeks for 2 years. Subjects with foot lesions were referred to their healthcare provider for treatment. In this cohort, 11 participants required lower limb amputation (rate 13.8 per 1000 person-years). Pivotal event analysis revealed that only one amputation was related to footwear, six amputations were due to non-footwear-related minor environmental trauma, two were due to progression of vascular disease (dry gangrene from critical ischemia), one was due to a self-care injury while cutting the toenails, and one was due to a decubitus ulcer. Previously proposed strategies to reduce the amputation rates in individuals with diabetes have focused heavily on footwear and education. However, even with this emphasis, amputation rates in the United States are still high. This study suggests that the prevention of minor environmental trauma, including household accidents, merits additional attention. We believe that further efforts to reduce amputation rates for individuals with diabetes will need to emphasize the prevention of minor trauma, especially in those already compromised with neuropathy and vascular disease. PMID- 14524520 TI - Topography of the distal tibial nerve and its branches. AB - The tibial nerve trunk and its branches were dissected in 20 embalmed cadaver legs and the relative topographic anatomy was defined at 3-cm intervals up to 15 cm proximal to the medial malleolar-calcaneal (MMC) axis. Each nerve branch was found in various locations. The calcaneal nerve was found to descend from medial to posteromedial. It was never found anterolaterally and only rarely laterally. The lateral plantar nerve was found to rotate externally from lateral and posterolateral to lateral and posteromedial as it descends. This nerve was not found medially or anteromedially. The first branch of the lateral plantar nerve was indistinguishable from the trunk of the tibial nerve descending medially to between the lateral plantar and calcaneal nerves. The overall pattern of the medial plantar nerve was an internal rotation from anteromedial (proximal) to anterior (distally). It was not found posteriorly. The flexor hallucis longus motor branch was located an average of 17.9 cm (range, 10-24 cm) proximal to the MMC axis. Preliminary application of these data has facilitated surgical dissection and afforded an understanding of how tibial nerve trunk pathology correlates with clinical manifestations. PMID- 14524521 TI - Surgical treatment of acquired flatfoot deformity: what is the state of practice among academic foot and ankle surgeons in 2002? AB - PURPOSE: This study was performed to review the present approach to the surgical treatment of acquired flatfoot deformity among a cross section of academic foot and ankle surgeons in the United States. METHODS: A typical case of stage II posterior tibial tendon dysfunction was developed. The hypothetical patient was a healthy 62-year-old male with a symptomatic acquired flatfoot deformity with posterior tibial tendon dysfunction. Nonoperative management had failed. A series of academic foot ankle surgeons were surveyed to identify their proposed operative treatment. Each surgeon surveyed was affiliated with an orthopaedic residency program and was directly responsible for teaching orthopaedic residents. Each surgeon had foot and ankle patients as the major component of their practice. The overall response rate was 81% (104/128). RESULTS: Ninety seven percent (101/104) of respondents reported that they would employ some type of bony procedure in their surgical treatment. Eighty-eight percent (89/104) described techniques that would preserve the subtalar and talonavicular joints. These included a medializing calcaneal osteotomy in 73% (76/104), a lateral column lengthening in 41% (43/104), and a medial column stabilization (first tarsometatarsal and/or navicular cuneiform arthrodesis) in 15% (16/104). Twelve percent (12/104) reported that they would perform an arthrodesis of one or more of the hindfoot joints. Ninety-eight percent (102/104) of respondents reported that they would employ some type of soft-tissue procedure. Ninety-four percent (98/104) would augment the posterior tibial tendon, 53% (55/104) would formally repair the spring ligament, and 70% (73/104) would address a presumed equinus contracture. CONCLUSIONS: There is a wide variation in the approach to surgical treatment of stage II acquired adult flatfoot deformity among academic foot and ankle surgeons. Most surgeons employed a combination of bony and soft-tissue procedures that preserved the subtalar and talonavicular joints. PMID- 14524522 TI - Fourth brachymetatarsia treated with distraction osteogenesis. AB - BACKGROUND: This study investigated metatarsal lengthening by distraction osteogenesis for fourth brachymetatarsia in 22 metatarsals (16 patients). METHODS: From May 1997 to May 2000, lengthening was performed with a monoexternal fixator, and distraction was started at a rate of 0.5 mm per day after a latency period of approximately 7-10 days. RESULTS: The average gain in length was 16.5 mm (range, 13-21 mm), equivalent to an increase of 39% (range, 28-51%), and the average healing index was 72.9 days/cm (range, 51.7-95.7 days/cm). The American Orthopaedic Foot and Ankle Society (AOFAS) average score for lesser toe was 86.3 (range, 47-100). The most common residual complication was subluxation of metatarsophalangeal (MTP) joint in five cases, with partial or total stiffness of the MTP joint. These complications happened in the group of metatarsals excessively lengthened more than 40% and made the AOFAS score poorer. The other complications were three cases of angular deformity in the lengthened bone, and two cases of pin-tract infection. CONCLUSION: Although distraction osteogenesis is an effective method to address fourth brachymetatarsia, stiffness or subluxation of the MTP joint was not uncommon. To avoid complications that can happen as a result of excessive lengthening, careful preoperative radiographic measurement to calculate the optimal amount of lengthening may help us to avoid overlengthening and the complications that accompany it. PMID- 14524523 TI - Results of chronic Achilles tendinopathy surgery on elite and nonelite track athletes. AB - Twenty-seven track athletes/runners with chronic Achilles tendinopathy underwent a retrospective review of 37 surgical Achilles procedures performed between 1990 and 1999. There were 22 males and 5 females. Average age of the group was 36.6 years (range, 16-75 years). The mean "return to activity" (RTA) was 10.6 +/- 6.3 weeks. For elite athletes, RTA was 7.9 = 4.8 weeks. For nonelite athletes, RTA was 15.0 +/- 6.2 weeks, and was statistically higher (p = .003). There was no significant difference between males' and females' RTA. Average follow-up for the group was 4.7 years (range, 1-10 years). Runners on average ran more than 60 miles a week. There were 14 elite and 13 nonelite athletes, including four Olympians, five sub-4-minute milers, and four National Champions. Return to competition and "100%" were 25.0 and 25.4 weeks, respectively, for the elite group. Return to competition for the nonelite group was 27.0 +/- 11.0 weeks. Eighteen patients underwent peritenolysis with a mean RTA of 7.7 weeks. Of this group, four patients with concomitant bony procedures had significantly slower RTA as opposed to the 14 patients who had peritenolysis only (mean, 4.5 weeks). Four patients had Achilles debridement for mucoid degeneration, and a mean RTA of 12.8 weeks. Eleven patients had Haglund-type procedure (retrocalcaneal exostectomy) had a mean RTA of 15.1 weeks. Six Achilles tendocalcinosis repair patients had a mean RTA of 12.0 weeks. There were three reoperations, two of which were performed by the author. Using the Testa Achilles tendon surgery rating scale, this group of patients had 34 "excellent" and 3 "good" results. The patients who underwent a reoperation were able to achieve "good" or better results. PMID- 14524524 TI - Traumatic rupture of both peroneal longus and brevis tendons. AB - Injuries of peroneal tendons are rare. Diagnosis of traumatic rupture is often late and presents as chronic ankle instability. A case of a complete traumatic rupture of both peroneal longus and brevis tendons with acute clinical and radiological diagnosis is presented. Surgical repair was performed by direct end to-end suture on the 4th day after trauma, with excellent functional outcome at 1 year follow-up. PMID- 14524525 TI - RE: Method for manual reduction of displaced intra-articular fracture of the calcaneus: technique, indications and limitations, Omoto H, Nakamura K, Foot Ankle Int. 22(11):874-879, 2001. PMID- 14524526 TI - RE: The effect on ankle dorsiflexion of gastrocsoleus recession, Pinney SJ, et al., Foot Ankle Int. 23(1):26-29, 2002. PMID- 14524527 TI - Local and systemic chemotherapy with taurolidine and taurolidine/heparin in colon cancer-bearing rats undergoing laparotomy. AB - Experimental studies in the therapy of malignant abdominal tumors have shown that different cytotoxic agents suppress the intraperitoneal tumor growth. Nevertheless, a general accepted approach to prevent tumor recurrences does not exist. Following subcutaneous and intraperitoneal injection of 10(4) colon adenocarcinoma cells (DHD/K12/TRb), the influences of both taurolidine or taurolidine/heparin on intraperitoneal and subcutaneous tumor growth was investigated in 105 rats undergoing midline laparotomy. The animals were randomized into 7 groups and operated on during 30 min. To investigate the intraperitoneal (local) influence of either taurolidine or heparin on tumor growth, the substances were applied intraperitoneally. Systemic and intraperitoneal effects were evaluated after intravenous injection of the substances. Both application forms were also combined to analyze synergistic effects. Tumor weights, as well as the incidence of abdominal wound metastases, were determined four weeks after the intervention. In order to evaluate the effects of the agents, blood was taken to determine the peripheral leukocytes counts. Intraperitoneal tumor growth in rats receiving intraperitoneal application of taurolidine (median 7.0 mg, P = 0.05) and of taurolidine/heparin (median 0 mg, P = 0.02) was significantly reduced when compared to the control group (median 185 mg). The simultaneous instillation of both agents also reduced the intraperitoneal tumor growth (median 4 mg, P = 0.04), while the intravenous injection of the substances caused no local effect. In contrast, the subcutaneous tumor growth did not differ among all groups. In all groups, abdominal wound recurrences were rare and did not differ. Independent of the agents and the application form, the operation itself caused a slight leukopenia shortly after the operation and a leukocytosis in the following course. Intraperitoneal therapy of either taurolidine or in combination with heparin inhibits local tumor growth and abdominal wound recurrences in rats undergoing midline laparotomy. Neither the intraperitoneal nor the intravenous application or the combination of the two agents influenced the subcutaneous tumor growth. The substances did not alter the changes of peripheral leukocytes. PMID- 14524528 TI - Enhanced green fluorescent protein-transfection of murine colon carcinoma cells: key for early tumor detection and quantification. AB - Many animal models for metastatic colorectal cancer represent clinical manifestations just inaccurately. We introduce a novel mouse model for metastastatic colorectal cancer. In order to remain close to the clinical disease a syngenic murine colon carcinoma cell line (colon 26 cells) was transfected with enhanced green fluorescent protein (EGFP). The transfected cells maintain the highly malignant attributes of the wild-type cells. Following injection into the portal circulation of Balb/c-mice, liver metastases occur in the same time span. Using the fluorescent attributes of the transfected cells, an approximation of the tumor load in liver tissue can be achieved by fluorescence activated cell sorting (FACS) and fluoroscan analysis. Tumor cell load in liver tissue can be accurately measured by Northern blot and Western blot analysis of liver tissue containing EGFP-transfected colon cancer metastases (1250 cells/mg liver tissue and 1000 cells/mg liver tissue) respectively. Confocal microscopy and intravital microscopy confirmed the growth of tumor metastases, originating from the intravascular compartments. The presented animal model using EGFP-transfected colon 26 cells allows the detecting of tumor growth in vivo and post mortem, as well as an accurate quantification of the tumor load in the liver tissue. PMID- 14524529 TI - Activity of biphenyl matrix metalloproteinase inhibitor BAY 12-9566 in a human breast cancer orthotopic model. AB - Matrix metalloproteinases (MMPs) have been implicated in the invasion, metastasis, and angiogenesis associated with human cancer by mediating the degradation of extracellular matrix components. In this paper, we report data that show that BAY 12-9566, a novel inhibitor of MMPs, inhibits angiogenesis, tumor regrowth, and the growth of lung metastases. BAY 12-9566, at 15-100 microM, inhibited tubule formation by human endothelial cells in an in vitro model, but did not prevent the proliferation of endothelial and human breast cancer cells. In the MDA-MB-435 human mammary carcinoma xenograft model, in which the primary tumor is transplanted into the murine mammary fat pad, BAY 12-9566, administered daily at a dose of 100 mg/kg/day p.o. after resection of the primary tumor, inhibited local tumor regrowth by 58% without causing any toxic effect. In addition, BAY 12-9566 treatment inhibited the number and volume of lung metastases by 57 and 88%, respectively. These effects were highly correlated with the serum concentration of BAY 12-9566 at the end of treatment. The serum of the treated animals, harvested 24 h after the last treatment, and the tumor regrown at the site of tumor transplant in the treated animals, contained less protein with MMP-9 activity (as measured in a gelatin zymography assay) than the corresponding controls. However, no difference in the activity of MMP-2 was observed. Although all clinical trials in cancer involving BAY 12-9566 have been halted, this MMP inhibitor has never been used in clinical trials in breast cancer. These results suggest that the novel MMP inhibitor BAY 12-9566 maybe a useful and safe oral treatment for breast cancer, adjunctive to surgery. PMID- 14524530 TI - Inhibition of alpha(v)beta3 integrin reduces angiogenesis, bone turnover, and tumor cell proliferation in experimental prostate cancer bone metastases. AB - The growth of metastatic prostate cancer cells in the bone involves an intimate interaction between the tumor cells and various elements of the bone microenvironment, resulting in increased rate of bone turnover and rapid tumor growth. The alpha(v)beta3 integrin has been shown to play an important role in tumor growth and angiogenesis, and is known to be critical to osteoclast formation and activity. This study was designed to examine the role of alpha(v)beta3 expressed by cells native to the bone in the growth and pathogenesis of prostate cancer bone metastases. Human prostate cancer cells which do not express alpha(v)beta3 or alpha(IIb)beta3 integrins were injected directly into human bone fragments previously implanted subcutaneously in SCID mice (SCID-human-bone model). At the same time treatment with anti-beta3 antibody fragment (m7E3 F(ab')2) i.p. at 300 microg/dose 3 x per week was initiated and continued for 2 weeks. In this system, m7E3 F(ab')2 only recognizes human bone derived alpha(v)beta3. Antibody inhibition of alpha(v)beta3 integrin in vivo resulted in a specific reduction in the proportion of antigenically-human blood vessels within tumor-bearing bone implants (from 73.5% +/- 3.93 in controls to 17.74% +/- 5.64 in treated animals). Proliferation of the alpha(v)beta3-negative tumor cells was also reduced, although the overall vessel density was maintained by compensating mouse vasculature. Blockage of human bone-derived alpha(v)beta3 also significantly reduced the recruitment of osteoclasts in response to tumor cells, as well as degradation of calcified bone tissue. Together these observations confirm the importance of alpha(v)beta3 in bone metabolism and angiogenesis, and point to the role of these processes in controlling growth of metastatic prostate cancer cells in the bone. PMID- 14524532 TI - Reduced angiogenesis in peritoneal dissemination of gastric cancer through gelatinase inhibition. AB - Marimastat is a broad-spectrum matrix metalloproteinase (MMP) inhibitor that inhibits almost all major MMPs, key enzymes in gastric cancer invasion and metastasis. We investigated the ability of marimastat to inhibit tumor angiogenesis in the severe combined immuno-deficient (SCID) mouse/human gastric cancer model of peritoneal dissemination. A human stomach adenocarcinoma cell line, TMK-1, was injected intraperitoneally into SCID mice. On the 7th day after tumor inoculation, the administration of marimastat (27 mg/kg/day) was initiated and the treatment was continued for 2 weeks using subcutaneously-inoculating mini osmotic pumps. On the 21st day, the mice were killed and the disseminated nodules were evaluated. Total weights, numbers, and the microvascular density of the disseminating nodules were significantly lower in mice treated with marimastat compared to the control group. Film in situ zymography demonstrated that net gelatinolytic activity in the tissues was weaker in treated-group nodules than in control-group nodules. Thus, our results suggested that marimastat inhibited peritoneal dissemination of human gastric cancer cells through inhibition of tumor angiogenesis, possibly involving the down-regulation of gelatinases, in SCID mice injected with human gastric cancer cells. PMID- 14524531 TI - c-Met tyrosine kinase receptor expression and function in human and canine osteosarcoma cells. AB - To further characterize the role of hepatocyte growth factor-scatter factor (HGF SF) and its receptor (c-Met) in osteosarcoma (OS), human OS cell lines with low (SAOS-2) and high (SAOS-LM2) metastatic potential, and cell lines derived from spontaneous canine OS were studied. All cell lines were evaluated for c-Met and HGF-SF expression and receptor activation using Northern, RT-PCR, and Western blot analyses, respectively. Functional activity of receptor-ligand interaction was measured using c-Met phosphorylation status, proliferation assays (anchorage dependent and -independent), Matrigel invasion, modulation of urokinase plasminogen activator (uPA) expression, and cell dispersion (scattering). All cell lines exhibited steady-state mRNA expression of c-Met. The canine OS cell lines also expressed HGF-SF mRNA as determined by RT-PCR analysis. Western analysis showed c-Met protein expression and HGF-stimulated (human) or constitutive (canine) receptor autophosphorylation. Treatment with recombinant human HGF resulted in enhanced proliferation in 3 of 5 OS cell lines and enhanced colony formation in 2 of 5 OS cell lines. Matrigel invasion was significantly enhanced in 3 of the cell lines and uPA levels were significantly increased in the SAOS-2 cells following HGF treatment. Scattering was enhanced in both the SAOS-2 and SAOS-LM2 cells. These data support the involvement of c-Met and HGF-SF in the growth and progression of human and canine OS, and may offer new targets for the development of therapeutic strategies for OS. PMID- 14524533 TI - Differential expression of osteopontin and bone sialoprotein in bone metastasis of breast and prostate carcinoma. AB - Breast and prostate cancer often metastasise to the skeleton. Interestingly, the histopathological characteristics of the bone lesions that arise from these two cancer types differ. Breast tumours give rise to metastases in the skeleton with a mixed lytic/sclerotic pattern, whereas a predominantly sclerotic pattern is seen in metastases from prostate tumours. Osteopontin (OPN) and bone sialoprotein (BSP) are bone matrix proteins that have been implicated in the selective affinity of cancer cells for bone. In the present study, 21 patient cases with skeletal metastasis and their respective primary tumours (12 with breast cancer, 9 with prostate cancer) were investigated by immunohistochemistry in order to assess the level of OPN and BSP. Moderate to strong OPN expression was found in 42% of all breast tumours and in 56% of all prostate tumours. Significantly more breast cancer bone metastases exhibited high OPN expression, 83%, as compared with prostate tumour bone metastases, 11% (P = 0.0019). In contrast, moderate to strong BSP expression was found in 33% of breast tumours and in 89% of prostate tumours. In the bone lesions, only 33% of breast tumour metastases showed moderate/strong BSP expression compared to 100% of prostate tumour metastases (P = 0.0046). This divergent pattern of OPN/BSP expression could be an important determinant for the different characteristics of these two types of bone metastasis, i.e., lytic vs. sclerotic, consistent with the proposed role of OPN in differentiation and activation of osteoclasts and of BSP as a stimulator of bone mineralisation. PMID- 14524534 TI - Different effects of constitutive nitric oxide synthase and heme oxygenase on pulmonary or liver metastasis of colon cancer in mice. AB - It has recently been reported that not only endogenous nitric oxide (NO) but also carbon monoxide (CO) produced by heme oxygenase (HO) have many physiological functions. The objective of the present study was to determine whether endogenous NO or CO is involved in the experimental pulmonary or liver metastasis of colon cancer in mice. Intravenous or intrasplenic injection of colon 26 cells from a mouse colon adenocarcinoma cell line resulted in multiple pulmonary or liver metastases. NG-nitro-L-arginine methyl ester (L-NAME), a competitive inhibitor of NO synthase (NOS), or zinc deuteroporphyrin 2, 4-bis glycol (ZnDPBG), a competitive inhibitor of HO, was administered to the mice only on the day of tumor inoculation. We assessed the number of tumor cells 24 h later and the outcome of metastases of the target organ. In the pulmonary metastasis model, L NAME increased both the number of tumor cells 24 h later and outcome of metastases 18 days later, but did not have a significant effect on liver metastasis. On the other hand, metastasis to the liver, but not that to the lung, increased following administration of ZnDPBG. These results suggest that the activities of NOS and HO could influence experimental metastasis in an organ specific manner. PMID- 14524535 TI - Continuous real time ex vivo epifluorescent video microscopy for the study of metastatic cancer cell interactions with microvascular endothelium. AB - Recent studies suggest that only endothelium-attached malignant cells are capable of giving rise to hematogenous cancer metastases. Moreover, tumor cell adhesion to microvascular endothelium could be crucial in metastasis predilection to specific organs or tissues. However, the existing in vitro and in vivo techniques do not provide for sufficient delineation of distinct stages of a dynamic multi step intravascular adhesion process. Here we report the development of an experimental system allowing for prolonged continuous ex vivo real-time observation of malignant cell adhesive interactions with perfused microvessels of a target organ in the context of its original tissue. Specifically, the vasculature of excised dura mater perfused with prostate cancer cells is described. An advantage of this technique is that selected fluorescently labeled tumor cells can be followed along identified vascular trees across the entire tissue specimen. The techniques provide for superior microvessel visualization and allow for uninterrupted monitoring and video recording of subsequent adhesion events such as rolling, docking (initial reversible adhesion), locking (irreversible adhesion), and flattening of metastatic cancer cells within perfused microvasculature on a single cell level. The results of our experiments demonstrate that intravascular adhesion of cancer cells differs dramatically from such of the leukocytes. Within dura microvessels perfused at physiological rate, non-interacting, floating, tumor cells move at velocities averaging 7.2 x 10(3) microm/s. Some tumor cells, similarly to leukocytes, exhibit rolling-like motion patterns prior to engaging into more stable adhesive interactions. In contrast, other neoplastic cells became stably adhered without rolling showing a rapid reduction in velocity from 2 x 10(3) to 0 microm/s within fractions of a second. The experimental system described herein, while developed originally for studying prostate cancer cell interactions with porcine dura mater microvasculature, offers great flexibility in adhesion experiments design and is easily adapted for use with a variety of other tissues including human. PMID- 14524536 TI - Seprase, a membrane-bound protease, alleviates the serum growth requirement of human breast cancer cells. AB - Seprase is a cell surface serine protease that is expressed to high levels by infiltrating ductal carcinomas of the breast but its function in malignancy is unknown. MDA-MB-435 (WT435) and MDA-MB-436 (WT436) human breast cancer cells express high levels of seprase as do the carcinoma cells in tumors of human breast cancer patients. To investigate its role in the pathobiology of breast cancer, seprase was specifically reduced in WT436 and WT435 cells by expression of antisense seprase cDNA. Decreased expression of seprase was confirmed in the antisense transfectants by zymography, immunoblotting, and fluorescence-activated cell sorting of cells labeled with antibody to seprase. Control-transfectants continued to express high levels of seprase. Seprase-deficient cells growing on type I collagen gels reveal a markedly different morphology than the parental or control-transfected cells that express high levels of seprase. The seprase deficient cells grow in islands and aggregates of tightly attached cells while cells with high seprase expression grow as groups of separate individual cells. Interestingly, the aggregated growth of the seprase-deficient cells was not correlated with increased expression of E-cadherin. Seprase-deficient breast cancer cells also exhibit altered growth properties. Seprase-deficient cells and those with high seprase levels proliferate in serum-containing media. However, in serum-free medium seprase-deficient cells proliferate much more slowly than their seprase-expressing counterparts. These findings indicate that seprase promotes the aberrant growth of breast cancer cells by reducing their dependence on exogenous growth factors. Seprase may contribute to the pathogenesis of breast cancer by promoting growth of the primary tumor and by facilitating the growth of breast cancer cells in metastases at other sites of the body. PMID- 14524537 TI - Breast cancer micrometastases: different interactions of carcinoma cells with normal and cancer patients' bone marrow stromata. AB - The apparently dormant breast cancer micrometastases in haemopoietic marrow are correlated with distant metastatic carcinoma dissemination. We studied in vitro interactions of carcinoma cells with adjacent stromata, using connective tissue cell cultures from breast and bone marrow samples of normal donors, comparing them to the pericancerous breast tissue and bone marrows of 12 selected patients with invasive breast carcinomas. Cancer cells were detected by immunocytochemistry and RT-PCR in all the bone marrows and in most blood samples of the studied patients. We monitored the growth and interaction of carcinoma MCF 7 cells with the stromata. The normal breast stroma sustained typical massive cancer growth. The pericancerous breast stroma induced the invasive mesenchymal pattern of growth. Normal bone marrow stroma induced the same conversion and was highly adhesive, retaining the cells in the stroma, but carcinoma patients' bone marrow stromata underwent low adhesive interactions with cancer cells, releasing them potentially into the circulation. The semi-quantitative RT-PCR indicated an enhanced expression of the hepatocyte growth factor and its receptor c-met in breast and bone marrow stromata of cancer patients. The input of cancer cells into the normal bone marrow may induce modifications of the local microenvironment, favourable for growth and release of carcinoma cells into the systemic circulation, which correlate with the poor prognosis of patients with bone marrow micrometastases. PMID- 14524538 TI - Arbovirus surveillance in Rhode Island: assessing potential ecologic and climatic correlates. AB - During 1995-2000, mosquitoes were collected from sites throughout Rhode Island and tested for the presence of arboviruses. Mosquito trapping was done weekly from June to October with CO2-baited light traps. In all, 186,537 mosquitoes belonging to 7 different genera were collected, of which Coquillettidia perturbans was most abundant. A total of 6,434 pools were processed for arbovirus isolation, from which 193 arboviral isolations were made. These included 109 Highlands J, 71 eastern equine encephalomyelitis, 1 California encephalitis serogroup, 2 Jamestown Canyon, 3 Cache Valley, and 9 Flanders viruses. Our isolations of Flanders virus represent the 1st reported occurrence of this virus in Rhode Island. After the 1999 sudden occurrence of the West Nile virus (WN) in the New York City area, a dead-bird surveillance program was started to test for this virus. Although no isolations of WN were made from mosquitoes, 87 virus isolations were made from a total of 330 wild birds tested. All the WN-infected birds were either American crows or blue jays. Isolation of WN from dead birds marked the 1st documented appearance of this virus in Rhode Island. Significant interannual variation of arbovirus activity in Rhode Island prompted us to examine if climate-associated factors such as rainfall and temperature correlate with virus activity. Total rainfall amounts from May to June were higher than normal in 1996 and 1998. These years showed significantly higher arbovirus activity. Deviations from normal temperature showed low correlation with arbovirus activity during the 6-year study period. Therefore, precipitation appeared to be more important than temperature in predicting arbovirus activity in Rhode Island. PMID- 14524539 TI - A geospatial study of the potential of two exotic species of mosquitoes to impact the epidemiology of West Nile virus in Maryland. AB - We used geospatial techniques to study the potential impact of 2 exotic mosquitoes, Aedes albopictus and Ochlerotatus japonicus japonicus, on the epidemiology of West Nile virus in Maryland. These 2 species have established populations in Maryland over the past 15 years. Larvae of both mosquito species are found in natural and artificial water-holding cavities and containers, particularly water in tires. Therefore, we used locations of licensed tire dealers and of tire dumps scheduled for clean up as an index for potential sources of mosquito vectors. This index was expected to underestimate the actual population of source habitats. West Nile virus activity in Maryland during 1999, 2000, and 2001 was indicated by the presence of dead, infected birds, particularly American crows and other corvids; infected pools of mosquitoes; and human and horse infections. Adult females of both mosquito species are aggressive, opportunistic feeders that have been observed to take blood meals from avian and mammalian hosts. Susceptible vertebrate hosts, particularly birds, are ubiquitously distributed throughout the developed areas of the state. This analysis demonstrated a spatial convergence of the virus, the exotic mosquito vectors, and susceptible hosts. This conjunction indicated that these 2 mosquito species have a high potential to serve as bridge vectors and thus, impact the epidemiology of West Nile virus under favorable environmental and climatic conditions. Positive mosquito pools were collected from only the Baltimore Washington metropolitan corridor, suggesting a newly created enzootic focus for this virus. Land-cover analysis of the sites where virus activity had been detected showed predominantly developed land uses. Analyses of the environmental justice aspects (social, economic, and housing characteristics) of block groups with human West Nile fever cases or with positive mosquito pools were equivocal. Human cases seemed to occur in developed block groups with lower income levels. PMID- 14524540 TI - Surveillance for the dengue vector Aedes aegypti in Tobago, West Indies. AB - An island-wide house survey was conducted in January 2002 to determine the geographic distribution, container profile, and population density of the Aedes aegypti in Tobago, West Indies. The results showed the Ae. aegypti infestation levels were significantly different (P > 0.01) among the 4 districts, with greater infestation levels (P > 0.01) observed in the Northern and Windward districts than in the Central and Leeward districts. From the 50 towns in Tobago, houses were found positive in Delaford (21), Argyle (18), and Goodwood (14). representing 42.1% of the total number of positive houses in the Windward district (3,971 houses); Parlatuvier (15), Whim (14), Castara (12), and Bloody Bay (12), representing 62.3% of the total number of positive houses in the Northern district (3,087 houses); Calderhall (12), Mason Hall (11), and Government House (10), representing 46.5% of the total number of positive houses in the Central district (4,706 houses); and Lambeau (10), Bucco (6), and Bethel (6), representing 53.7% of the total number of positive houses in the Leeward district (3,175 houses). The majority (66 or 63.5%) of dengue cases occurred in the Central district where the Breteau indices ranged from 7.1 to 44.0 (mean = 16.6). These results suggest that a more systematic and sustained vector control program that uses both biological and chemical control methods should be adopted to reduce Ae. aegypti populations to below dengue transmission thresholds. PMID- 14524541 TI - Molecular and electron microscopic identification of Wolbachia in Culex pipiens complex populations from the Upper Rhine Valley, Germany, and Cebu City, Philippines. AB - We detected Wolbachia infections in Culex pipiens complex mosquitoes from the Upper Rhine Valley, Germany, and Cebu City, Philippines, with the use of polymerase chain reaction (PCR) amplification of the 16S rDNA of the bacteria and further confirmation by electron microscopy. Species of 5 different autogenous and anautogenous populations of Culex pipiens from Germany and 1 population of Culex quinquefasciatus from the Philippines were used in this study. Larvae were sampled from different localities and reared to the adult stage for further processing. Whole adult males and the heads of adult females were processed by PCR, whereas the abdomens of females were used for electron microscopic study when an infection was detected by PCR. All Culex populations assayed by PCR showed infection of Wolbachia at rates between 10 and 100%. Females from different populations exhibited higher infection rates than did the males, and, likewise, Cx. pipiens populations that were autogenous showed higher infection rates of Wolbachia than did anautogenous individuals. In Cx. quinquefasciatus populations, males were infected at 33.33% and females, only 10%. We studied the ultrastructure of Wolbachia in the ovaries of species belonging to Cx. pipiens complex by means of transmission electron microscopy. The bacteria exhibited typical morphology for Wolbachia with 3 enveloping membranes. PMID- 14524542 TI - Pre- and postprandial mosquito resting behavior around cattle hosts. AB - Spatial distribution of mosquitoes around a bovine host was studied in November 1997 in northern Thailand (17 degrees 38'N. 99 degrees 23'E). Forty bamboo stakes were arranged 1 m apart, in 4 rays of 10, around a cow tethered in an open field. All mosquitoes found resting on the stakes were collected by aspiration between 7:00 p.m. and 11:00 p.m., sexed, and identified to species; and feeding status was categorized as fed or unfed. Collections were repeated over 8 nights, with and without the host cow. A total of 1,566 mosquitoes from 25 species (5 genera) was collected. Anopheles aconitus was the most abundant species (643 individuals), followed by An. peditaeniatus. Culex vishnui, and Cx. pseudovishnui. We found that the number of mosquitoes collected from the stakes was related to the presence of the cow host: the number of mosquitoes collected was unrelated to the compass point location of the bamboo stakes, with the exception of Mansonia uniformis; unfed mosquitoes preferred bamboo-stake resting sites that were closer to the host; the daily fed to unfed ratio of the dominant species was negatively correlated with the daily total number of mosquitoes collected; and fed and unfed mosquitoes clustered in interspecific heterogeneous groups around the host cow. Cluster analysis separated the species into 2 groups. The 1st consisted of 5 species with higher proportions of fed mosquitoes, whereas the 2nd, represented by 7 species, aggregated around the host within a distance of 1-4 m with lower proportions of fed mosquitoes. The interspecific variation in the distribution of unfed females was presumed to be due to a lack or feeding success. We discuss the significance of prebiting resting. In cases in which large numbers of mosquitoes are present, prebiting resting can be adaptive to avoid host defensive behavior triggered by attacking mosquitoes. PMID- 14524543 TI - Quantitative evaluation of funnel traps for sampling immature Aedes aegypti from water storage jars. AB - The trend is increasing to incorporate assessments of abundance into surveys for immature Aedes aegypti to identify the most important types of containers that should be targeted for control. In this study, we examined whether funnel traps could be used to sample immature Ae. aegypti from water storage jars ranging in size from 0.28-m diameter (30 liters) to 0.52-m diameter (150 liters). The effects of jar size and duration of funnel trap sampling were investigated and a set of calibration factors was developed to convert funnel trap numbers to absolute population estimates (0.28-m diameter = 2.5, 0.38-m diameter = 3.0, 0.48 m diameter = 4.6, and 0.52-m diameter = 7.4). Although the funnel traps were highly sensitive (90-100%) for detecting immature Ae. aegypti at densities as low as 25 3rd and 4th instars per jar, the large variation in funnel trap recapture rates meant that absolute population estimates based on a single funnel trap sample were inaccurate. However, by using a computer simulation, estimates of the total overall numbers of larvae from multiple jars were reasonably accurate (+/- 20%), if more than 50 positive jars were surveyed. For example, 95% confidence intervals for the percentage error in estimated numbers of immatures from a series of 50 0.38-m-diameter and 50 0.52-m-diameter jars, were -10.0% to +10.2% and -19.9% to +17.8%, respectively. Although we generally recommend the use of nets to sample immature Ae. aegypti in jars, under some conditions funnel traps may be more acceptable than nets, because some householders object to the increased turbidity associated with net sampling in jars. PMID- 14524544 TI - Field and laboratory evaluations of potential oviposition attractants for Aedes albopictus (Diptera: Culicidae). AB - We tested five volatile synthetic chemicals (dimethyl disulfide, indole, 4 methylphenol, 3-methylindole, and trimethylamine) as potential oviposition attractants of Aedes albopictus in field and laboratory experiments. The 5 synthetic compounds were loaded into controlled-release packets, which consisted of a cellulose material sealed within a permeable plastic membrane, that were used to bait water-filled ovitraps at 5 field sites. Aedes albopictus exhibited no oviposition preference for any of the baited traps versus adjacent traps containing only water. In addition, there was no difference in the mean number of eggs laid per trap-day by Ae. albopictus among ovitraps treated with the five compounds. We conducted behavioral bioassays to determine if the lack of response to the putative oviposition chemicals in the field was due to a concentration effect. A binary sticky-screen bioassay was used to measure attraction of gravid females to olfactory stimuli. Compounds were evaluated over a range of concentrations that spanned 3-5 logs (0.0083 to 8.3 or 83 mg/liter). Three concentrations of 4-methylphenol (0.083 mg/liter, 0.83 mg/liter, and 8.3 mg/liter) and 1 concentration of 3-methylindole (8.3 mg/liter) were significantly repellent. All other concentrations of the 5 chemicals tested did not attract more females than did a water control. Electoantennography indicated that Ae. albopictus did not exhibit a physiological response to 0.25 ng of any of the five chemicals tested. Because Ae. albopictus did not exhibit attraction, greater oviposition. or an electrophysiological response to any of the compounds tested, these compounds do not appear to be effective lures for baiting ovitraps for surveillance or control of this mosquito. PMID- 14524545 TI - An adulticidal sticky ovitrap for sampling container-breeding mosquitoes. AB - The efficacy of a standard ovitrap and an ovitrap featuring an internal wall covered by a polybutylene adhesive was compared in field studies in Cairns, Australia. The sticky ovitrap was as effective as the standard ovitrap in detecting Aedes aegypti, with 67.5% and 64% of traps positive for Ae. aegypti, respectively. Significantly higher numbers of Ae. aegypti were collected by traps set outside rather than inside premises. Sticky ovitraps also readily collected Ochlerotatus notoscriptus and, especially, Culex quinquefasciatus. With a 10X hand lens, mosquitoes of these species could readily be identified in traps set for 3 and 7 days. The sticky ovitraps were comparable in cost and as time efficient as standard ovitraps. The greatest advantage of the sticky ovitrap is the collection of adult female mosquitoes, negating the need to rear larvae for identification and providing a faster, more direct measure of the effectiveness of ovipositional attractants than egg counts. Finally, we demonstrated that sticky ovitraps, being adulticidal, have potential as a supplementary control measure, especially for quarantine programs designed to prevent the import and export of container-breeding vector mosquitoes at sea- and airports. PMID- 14524546 TI - Optimizing an aerial spray for mosquito control. AB - The role of proper flight line positioning was demonstrated to be critical for maximizing product efficacy when spraying in crosswinds by small-droplet spray strategies. Characterization studies indicated clearly that aircraft height, small drop emission distribution, and the ambient winds combine to dilute the spray cloud before impacting surface targets. A 5-fold increase in mortality was achieved when we used optimization techniques to position the aircraft during a crosswind treatment test. PMID- 14524547 TI - Efficacy of Bacillus thuringiensis israelensis, Bacillus sphaericus and temephos for managing Anopheles larvae in Eritrea. AB - We evaluated the larvicidal activity of the granular formulation of Bacillus thuringiensis israelensis (Bti) serotype H-14 (Vectobac G, 200 ITU/mg) and Bacillus sphaericus (Bsph) serotype H5a5b (Vectolex CG, 670 Bs ITU/mg) against Anopheles arabiensis and other mosquitoes in breeding habitats in 3 sites, Gash Barka, Anseba, and Debub zones, in Eritrea. The primary objective was to determine the optimal application rate and duration of effect for Bti and Bsph in representative larval habitats as compared with the organophosphate temephos. The biolarvicides were tested at 100% (high) and 50% (low) of the maximum recommended application rate. Temephos was applied at a rate of 100 ml/ha. At least 4 replicate experiments with Vectobac G (5.6 and 11.2 kg/ha), Vectolex CG (11.2 and 22.4 kg/ha) were conducted in each study site. All 3 larvicides caused significant mortality of the main malaria vector species, An. arabiensis, and other mosquito species (Anopheles cinereus, Anopheles pretoriensis, Culex quinquefasciatus). The larvicidal activity for Bti and Bsph was variable depending upon breeding habitat, mosquito species, and general ecology of the area. Both biopesticides had a similar duration of activity (2-3 wk) and were generally as effective as temephos for these time periods. In some cases, the high and low application rates for Bti and Bsph produced equivalent control over 2-3 wk. The 2 Bacillus biopesticides were less effective in habitats with high algal content and in fast flowing streams primarily because of the inability to penetrate algal mats and dilution effect, respectively. The results show that application of the 2 biolarvicides bimonthly to streambed pools, rain pools, and similar habitats would maintain control of the anopheline mosquito population. PMID- 14524548 TI - A checklist of the mosquitoes of Idaho and two new state records: Ochlerotatus decticus and Ochlerotatus euedes. AB - This checklist of the mosquitoes of Idaho updates the list of Brothers (1971), adding 5 species. From the literature. 3, Ochlerotatus mercurator, Ochlerotatus nevadensis, and Ochlerotatus punctor, are added and 1, Ochlerotatus stimulans, is deleted. We are reporting 2 new state records, Ochlerotatus decticus and Ochlerotatus euedes, and extending their ranges westward in the United States. One new genus and 2 new subgenera are also added. PMID- 14524549 TI - Suggesting new species? Comments on "Evidence for a new species of Anopheles minimus from the Ryukyu Archipelago, Japan". AB - Species recognition and identification are crucial for the planning and evaluation of vector control. In recent years, considerable effort has been made to clarify the species composition of vector taxa. The danger exists, however, that new species are suggested on little evidence and that intraspecific variation is insufficiently surveyed. We discuss these topics on the basis of recent studies on the Anopheles minimus complex, one of the most widespread vector groups in Southeast Asia. PMID- 14524550 TI - Efficacies of the mosquitomagnet and counterflow geometry traps in North Queensland, Australia. AB - We conducted three trials near Cairns, Australia, to compare the numbers of mosquitoes collected with the standard Encephalitis Vector Surveillance (EVS) and Centers for Disease Control (CDC) light traps with the new MosquitoMagnet (MM) and counterflow geometry (CFG) traps with the use of a 4 x 4 latin square experimental design. The MM was generally equal to the performance of the CDC and CFG traps, ranking ahead of or equal to one or both traps in 2 of 3 trials, although there were no significant differences in the performances of the MM, CDC, and CFG traps. The EVS trap ranked last in all trials. The MM, being self powered and self baited via combustion of propane for up to 20 days without requiring a propane refill, would be suitable for collection of adult mosquitoes in remote areas that do not have access to dry ice. PMID- 14524551 TI - Laboratory evaluation of toxicity of 16 insect repellents in aerosol sprays to adult mosquitoes. AB - Sixteen commercial insect repellents (6 botanical and 10 synthetic organic products) in spray formulations were evaluated in the laboratory for adult knockdown (KD) and mortality of laboratory-reared female Aedes aegypti, Aedes albopictus, and Anopheles quadrimaculatus. All tested products produced significant posttreatment KD and 24 h mortality of all 3 mosquito species. In our study, the synthetic organic repellents induced faster KD and KD of higher magnitude in adult mosquitoes than the botanical product repellents except geraniol-based MosquitoSafe. All tested formulations except 2 botanical repellent products caused 100% 24 h mortality of Ae. aegypti and all but 1 caused 100% 24 h mortality of Ae. albolpictus and An. quadrimaculatus. PMID- 14524552 TI - Comparative toxicity of permethrin- and bifenthrin-treated cloth fabric for Anopheles farauti and Aedes aegypti. AB - In this laboratory study, we applied 3 formulations of permethrin (Peregin 500 EC, Dragnet 500 EC, and Dragnet 100 ME) and 2 of bifenthrin (Biflex 10 ME and Talstar 80 SC) to swatches of Australian military shirt fabric. The knockdown and mortality of Anopheles farauti and Aedes aegypti after exposure to treated fabrics were compared. The mortality of An. farauti exposed to permethrin-treated swatches for 3 min in World Health Organization test kits was 94.2-100% after initial treatment and fell to < 28% after 2 cold water washes. and knockdown was < 20% after 3 washes. The mortality of An. farauti exposed to bifenthrin-treated swatches was initially 100% and remained > 55% after 3 washes, whereas knockdown was < 25% after 2 washes. Mortality of Ae. aegypti exposed by tarsal contact to permethrin- and bifenthrin-treated fabrics was 84.8-100% prior to washing and fell to < 21% and < 40%, respectively, after 1 cold water wash. The ability of Ae. aegypti to obtain a blood meal through treated fabrics was variable, and a small percentage (0-6.1%) of mosquitoes obtained a blood meal through fabrics after initial treatment. The effect of cold water washing on the persistence of both chemicals in fabric by chemical assays showed that between 58% and 66% of both chemicals was lost from the test fabric after a single wash. PMID- 14524553 TI - Restating the obvious: the world can't afford American health care. PMID- 14524554 TI - A multibillion dollar industry, but not for poor countries. PMID- 14524555 TI - Reading and writing in the field of mechanical circulatory support. AB - Today there is a multiplicity of options for publishing scientific research work. In a specialized field like blood pump studies it is readily established that a very small number of journals publish the vast majority of work. Reading and publishing in those journals will provide the quickest exposure of the work to the most specifically interested and relevant people, even if the "impact factors" of those journals do not have stratospheric values. PMID- 14524556 TI - The beginning of clinical hemofiltration: a personal account. PMID- 14524557 TI - What price support? Ventricular assist device induced systemic response. AB - Use of ventricular support systems has been associated with myriad systemic complications. Engendered by the blood-biomaterial interface of a unique host/device relationship, these complications include diverse humoral dyscrasias that frequently culminate in episodes of bleeding, hemolysis and thrombogenicity, heightened susceptibility to inflammation and infection, and transient immunal compromise. Recent endeavor in biocompatibility research has served to illustrate the critical role played by cellular, humoral, and neurohormonal components in regulating cytokine expression and has provided insight into the complexities involved in such biomechanical juxtapositions. The following is intended as a review of current literature attempting to address the many aspects of this host/device interaction and their consequences for the supported patient. PMID- 14524558 TI - Intraperitoneal transplantation of pancreatic anlagen. AB - To determine whether embryonic pancreatic anlagen transplanted to an intraperitoneal site in adult hosts grow, differentiate, and function, we implanted pancreas from embryonic day (E) 12.5 Lewis rat embryos into the omentum of adult Lewis rats or C57Bl/6J mice. E12.5 pancreatic anlagen were relatively undifferentiated except for the presence of condensing tubuloacinar cords. By 2 weeks after implantation, pancreatic anlagen transplanted into rats had enlarged and differentiated such that islets of Langerhans that stained positive for insulin could be delineated. Continued differentiation, as reflected by the presence of "ductal" islets connected to the duct epithelium, was observed at 6 weeks after implantation. At 15 weeks after implantation, "mature" islets had separated from the ducts. Electron microscopy showed eccentric dense bodies within clear vacuoles consistent with insulin granules. Little or no acinar tissue was present in developed anlagen. Within 5 weeks of pancreatic anlagen transplantation, levels of glucose in rats rendered diabetic by an injection of streptozotocin were normalized compared with levels in nontransplanted diabetic controls. Rat pancreatic anlagen underwent growth and development in the peritoneum of C57Bl/61 mice that received costimulatory blocking agents but not in the absence of costimulatory blockade. We concluded that whole E12.5 pancreatic anlagen undergo growth, differentiation, and function after intraperitoneal placement. Implantation of the embryonic pancreas, a "cellular" transplant, is followed by selective differentiation of islet compared with acinar components. PMID- 14524560 TI - Polyethylene glycol additives reduce hemolysis in red blood cell suspensions exposed to mechanical stress. AB - Mechanical damage to blood cells is of considerable concern in the development and use of circulatory assist devices and other blood contacting systems. Furthermore, hemodilution with saline, dextran, and other plasma expanders applied during extracorporeal circulation and dialysis increases red blood cell (RBC) susceptibility to the high shear stresses associated with these procedures. In this paper, we present polyethylene glycol (PEG) as a potential erythrocyte protective agent against mechanically induced cellular trauma. Bovine RBCs were subjected to mechanical stress induced by rolling stainless steel shots through RBC suspensions for a constant exposure time. The suspensions were prepared at a hematocrit of 30% in various media: PEG (20,000 molecular weight), autologous bovine plasma, Dextran 40 solution, and phosphate buffered saline (PBS). RBC suspensions in Dextran 40 were prepared at a viscosity similar to the PEG suspensions. We found the hemolysis level of RBCs suspended in plasma and in PEG solutions to be several times lower (p < 0.001) than in the Dextran and PBS solutions. No statistically significant difference was found between the hemolysis that occurred in suspensions of RBCs in autologous plasma and in 2.0% PEG solutions. Even PEG concentration as low as 0.1% reduced hemolysis by more than 40% compared with PBS or the same concentration of Dextran in suspension medium. Our data demonstrate the efficacy of PEG molecules in reducing mechanical trauma to erythrocytes and suggest the potential for using PEG in assisted circulation, dialysis, and other procedures where RBCs are subjected to extensive mechanical stress. PMID- 14524559 TI - Experimental study of bone morphogenetic proteins-2 slow release from an artificial trachea made of biodegradable materials: evaluation of stenting time. AB - We manufactured an artificial trachea that slowly releases bone morphogenetic protein 2 (BMP-2) and used it to replace a section of the canine trachea. We made a three-layered prosthesis composed of an outer layer of gelatin sponge, a middle layer of collagen sponge, and an inner silicone tube. BMP-2 solution was soaked into the gelatin sponge layer. An approximately 3 cm length of the canine trachea was resected, and the artificial trachea was inserted into the resulting gap and anastomosed. The implanted portion was covered by periosteum. At 2, 4, and 8 weeks after surgery, the inner silicone tube was removed. Soon after removal of the silicone tube at 2 and 4 weeks, the dogs died of choking because of collapse of the trachea. One dog whose silicone tube was removed at 8 weeks was able to survive without choking. At 6 months after removal of the silicone tube, the bronchoscopic findings revealed that the gap in the trachea had been closed by regenerated tissue and covered by mucosa. We have demonstrated that our artificial trachea slowly releasing BMP-2 requires at least 8 weeks to achieve regeneration of solid tissue to support the tracheal gap. PMID- 14524561 TI - On-line clearance: a useful tool for monitoring the effectiveness of the reuse procedure. AB - Reprocessing of a dialyzer for repeated use in the same patient is widely practiced. The dialyzer fiber bundle volume (FBV) is monitored as an indicator of the dialyzer's suitability for continued use, with standards for reprocessed dialyzers requiring a FBV of greater than 80% of a new dialyzer to be maintained. We have used on-line measurement of clearance of sodium (OLC module, Fresenius Medical Care, Walnut Creek, CA) to assess small molecule clearance changes during and between treatments for a group of 29 chronic hemodialysis patients who reused high flux polysulfone dialyzers (F80, Fresenius Medical Care, Lexington, MA) reprocessed using citric acid and heat (95 degrees C). Data pertaining to the initial, 5th, 10th, and 15th uses were analyzed and showed that, within a single dialysis session, there was a trend for the clearance to reduce throughout the treatment (p < 0.001). Overall, there was also a trend for clearances to decline with increasing number of reuses (p < 0.008). Changes in FBV occurred, but such changes remained within the guidelines suggested by standards. It is concluded that on-line clearance measurements provide a simple noninvasive method to monitor dialyzer performance over each use and between uses. PMID- 14524563 TI - Effect of exercise on the diameter of arteriovenous fistulae in hemodialysis patients. AB - Physicians routinely recommend hand squeezing exercises for end-stage renal disease patients with newly placed arteriovenous fistulae (AVF) to increase the rate of fistula maturation. However, this practice has never been shown to actually work. To determine whether hand squeezing has an acute effect on fistula diameter, we examined 23 patients with newly created AVF (1 week to 10 months before study, mean 2.8 months). Using duplex ultrasound, we measured the diameter of the fistula three times before and three times after 5 min of squeezing a rubber ball. Fistula diameter increased in 20 of 23 patients; the mean change in fistula diameter was 9.3% (p < .0001). These data suggest that fistulae do dilate acutely after hand squeezing exercise and that this exercise should continue to be recommended. PMID- 14524562 TI - Experience with the JMS fully automated dialysis machine. AB - A fully automated dialysis machine has been developed and evaluated clinically. It uses highly pure dialysate (produced by a new dialysate cleaning system) instead of the conventional physiologic saline for the processes of priming, guiding blood to the dialysis machine, replenishing fluid, and returning the blood to the body. The piping for the dialysate is in the shape of a loop, and the dialyzer coupler has no mechanical parts that might become contaminated. As a result of these and certain other improvements in machine design, it is now possible to obtain reasonably clean dialysate. For the priming process, the machine uses a volume of up to 4 L of the dialysate after reverse filtration from the dialyzer. Most foreign matter or eluates can be removed from the dialyzer and the blood channels. Before blood is guided out of the body into the dialysis system, the needles inserted in the artery and vein are simultaneously connected to the blood channel, and the dialysate remaining in the channel is removed from the dialyzer. If the patient's blood pressure falls during dialysis, the dialysate can be replenished at any desired flow rate for reverse filtration. Blood return can be started automatically when the planned dialysis time has elapsed and the target water volume has been removed. The cleaned dialysate is infused from the dialyzer into the blood channel by reverse filtration to allow the blood to be returned to the body via both the artery and the vein at the same time. A total of 216 units of this fully automated dialysis machine have been placed in service at two of our facilities. During the 6 month period beginning in July 2001, they were used for 40,000 hemodialysis sessions in 516 patients. During the dialysate preparation process, the endotoxin levels in the reverse osmosis (RO) water, prefilter dialysate, and reverse filtered dialysate were all less than 1 EU/L. The time required to guide blood into the dialyzer (n = 39) decreased from the 4.6 +/- 1.4 minutes with the conventional machines to 3.2 +/- 0.6 minutes with the new machine (p < 0.01). The time required to return blood to the body also decreased from 8.6 +/- 2.2 minutes with the conventional machines to 6.8 +/- 0.7 minutes with the new machine (n = 34). No mechanical trouble was encountered with the fully automated dialysis machine units during the 40,000 hemodialysis sessions, and the workload of the dialysis unit staff in terms of the time needed to guide out and return blood to the body was significantly reduced. Because the machine simplifies the maneuvers required during hemodialysis, it is expected to contribute greatly to preventing medical accidents and in hospital infections associated with hemodialysis. PMID- 14524564 TI - Development of a photolytic artificial lung: preliminary concept validation. AB - There is an established need for pulmonary technology capable of facilitated gas exchange in the blood, thereby bypassing the alveolar-capillary interface. To address this need, we emulated one of the best-known photolytic reactions in nature, photosynthesis, in which green plants use sunlight to drive the exchange of oxygen for carbon dioxide. Our goal in the current study was to demonstrate the feasibility of direct photolytic conversion of water to liquid phase oxygen (dissolved oxygen [DO]) in synthetic serum. To this end, we constructed a test flow cell consisting of a conductive coating of vacuum-deposited titanium (Ti) metal, adherent TiO2 (anatase), and MnO2, applied as a laminate to a glass substrate, and then immersed the device in Locke's-Ringer solution (synthetic blood serum). Long wavelength (low energy) ultraviolet A laser light, directed to the transparent glass slide, reproducibly resulted in the generation of an active form of oxygen (AO), which was subsequently converted directly by the catalytic action of MnO2 to DO. The absence of light activation provided an entirely null response. We conclude that the photolytic production of DO from water in a blood serum surrogate, with commensurate CO2 clearance, is feasible. A prototype photolytic module is proposed, which uses multiple parallel photolytic surfaces to improve system production capacity and CO2 clearance through selective gas liquid separation from the oxygen-enriched fluid. PMID- 14524565 TI - Toward ambulatory arteriovenous CO2 removal: initial studies and prototype development. AB - Extracorporeal arteriovenous carbon dioxide removal (AVCO2R) using percutaneous cannulae and a low resistance gas exchanger achieves near total CO2 removal, allowing lung rest and potentially improving survival. AVCO2R, redesigned to allow ambulation, has potential as treatment for severe chronic obstructive pulmonary disease or rehabilitation before lung transplant. The purposes of this study were to 1) determine the optimal ambulatory access for AVCO2 removal and 2) develop a prototype Ambulatory-AVCO2R gas exchanger. Initially, reinforced Gore Tex 6 mm (two) and 8 mm (four) grafts were anastomosed to sheep carotid arteries and jugular veins as a loop in parallel to the cranial circulation to determine blood flow capabilities. Blood flow was 100-150 ml/min with a 14 gauge dialysis needle, and transected 6 mm Gore-Tex grafts achieved 500-900 ml blood flow, whereas transected 8 mm grafts achieved up to 2000 ml/min flow. The polytetrafluoroethylene (PTFE) loops were then connected to our newly developed ultra low resistance pumpless gas exchanger for ambulatory AVCO2R. The average pressure gradient across the prototype Ambulatory-AVCO2R gas exchangers (n = 5) was 2.8 +/- 0.8 mm Hg, and mean CO2 removal was 104.8 +/- 14.0 ml/min, with an average blood flow of 900 ml/min. We conclude that an 8 mm Gore-Tex reinforced graft arteriovenous loop supplies ample blood flow for our new ultra low resistance Ambulatory-AVCO2R to achieve near total CO2 removal. PMID- 14524566 TI - Venovenous extracorporeal membrane oxygenation for respiratory failure in inotrope dependent neonates. AB - It is often stated that venovenous extracorporeal membrane oxygenation (VV ECMO) should not be used in inotrope dependent patients. It is our practice to use VV ECMO in most patients with respiratory failure even though many of these patients are receiving significant doses of inotropes. Our objective was to review the mode of ECMO in relation to precannulation doses of inotropes administered to neonates treated with ECMO for respiratory failure. Forty-three consecutive case notes were reviewed. Data were collected for basic demographic and ECMO parameters. Inotropic doses were converted to a single score for ease of comparison, with one point equivalent to 1 microg/kg/min dopamine. Forty-three neonates were studied; 37(86%) were treated with VV ECMO and 6 (14%) were treated with VA ECMO. Significant pre-ECMO inotropic support (score > 10) was present in 30 (70%) of the 43 cases. Of these patients, 26 were treated via VV ECMO with a survival rate of 84%, while 4 were treated with VA ECMO with a survival of 75%. Inotrope scores fell to nonsignificant levels (< 10) within 24 hours, regardless of ECMO mode. Mean arterial blood pressure remained above precannulation levels in both groups. VV ECMO allows safe treatment of neonatal respiratory failure in the presence of significant inotropic support. We recommend VV ECMO for neonatal respiratory failure in all cases except where double lumen cannulation is impossible or when septic shock is refractory to inotropic support (i.e., mean blood pressure < 35 mm Hg despite inotrope score of > 100). PMID- 14524567 TI - Linear performance characteristics of latissimus dorsi muscle: potential for cardiac assistance. AB - Knowledge of the quantitative performance capabilities of skeletal muscle in a linear geometry is necessary to predict the performance and to optimize the design of linearly configured, skeletal muscle powered cardiac assist devices (MCADs). This study determined the performance characteristics of goat latissimus dorsi muscle (LDM) using a linear, ex vivo experimental apparatus. In five goats, the LDM (130.6 +/- 18.8 g) was dissected free of its distal attachments, connected to a series of weights (500 to 2250 g), and maximal tetanic contractions were elicited via thoracodorsal nerve stimulation. Second order polynomial equations were derived to represent each of the following variables versus load: muscle prestretch (range 1.5 to 5.4 cm), contraction duration (220 to 360 milliseconds), contraction shortening distance (13.5 to 10.9 cm), contraction velocity (60 to 31 cm/s), generated stroke power (3 to 7 W), and stroke work (0.7 to 2.4 J). Analysis of the potential stroke volumes obtained with a linearly configured, cylindrically shaped MCAD directly coupled to the circulation indicate that a feasible MCAD operating region exists based on the LDM performance data across a range of device geometries and mean ejection pressures. PMID- 14524568 TI - In vivo evaluation of the NEDO biventricular assist device with an RPM dynamic impeller suspension system. AB - Since 1995, the Baylor College of Medicine group has been developing the NEDO Gyro permanent implantable (PI) pump. The Gyro PI pump has achieved outstanding results up to 284 days with no thrombus formation during the left ventricular assist device (LVAD) animal experiments. However, in biventricular assist device (BVAD) animal experiments, thrombus formation did occur. An in vitro experiment showed the reason for thrombus formation was caused by the missed magnetic balance between the impeller and the actuator. On the basis of this result, the revolutions per minute (RPM) impeller suspension system was developed. Six long term animal studies were performed in bovine models. Survival periods were 90, 80, 60, 51, 48, and 37 days, respectively. No thrombus was observed in the pumps with the exception of one right pump. In that experiment, the thrombus formation may have occurred when the pump had a low flow because of outflow kinking. In this article, the antithrombogenic effect of this RPM impeller suspension system will be discussed. PMID- 14524569 TI - Preliminary experiment with a newly developed double balloon, double lumen catheter for extracorporeal life support vascular access. AB - Recently, venovenous extracorporeal life support (VVECLS) using a double lumen catheter has been clinically used to avoid neurologic complications in the treatment of respiratory failure for neonates. However, recirculation, which is a limiting factor for oxygen delivery, still exists, and thus it does not contribute to oxygenation of the patient. We developed a newly designed double lumen catheter with a double balloon (DBDL) catheter for ECLS vascular access and performed two animal preliminary experiments in normal and hypoxic dog models (normal ventilation and one lung ventilation experiments) to investigate whether the DBDL catheter could prevent recirculation and maintain oxygen delivery to systemic circulation. The DBDL catheter (JCT Co., Hiroshima, Japan) of 15 Fr was fabricated from silicone. It consists of two lumens for drainage and return of blood with two balloons (distal and proximal balloons) that prevent oxygenated blood mixing with unoxygenated blood. VVECLS using a DBDL catheter was performed in 13 mongrel dogs (8 dogs for normal ventilation experiment weighing 12.9 +/- 1.6 kg [mean +/- SD], 5 dogs for one lung ventilation experiment weighing 16.6 +/ 2.5 kg [mean +/- SD]) under anesthesia in the two experiments. The bypass flow ranged from 10-40 ml/kg per minute in the normal ventilation experiment. VVECLS in the one lung ventilation experiment was performed with maximal bypass flow for 6 hours (ranged from 25.2 +/- 8.0-28.3 +/- 8.7 ml/kg per minute at balloon inflation and deflation). Recirculation and oxygen transfer of artificial lung with or without balloon inflation during VVECLS were studied. Recirculation decreased with balloon inflation at varied bypass flows during VVECLS in the normal ventilation experiment (varied from 1.5 +/- 14.6-12.8 +/- 16.7%) and for 6 hours after VVECLS initiation in the one lung ventilation experiment (varied from 12.2 +/- 12.2-19.2 +/- 6.5%). In particular, the values at 3 and 6 hours were significantly lower than that of balloon deflation in the one lung ventilation experiment. The difference in O2 content between inlet and outlet in the artificial lung with balloon inflation was significantly higher than that of balloon deflation (varied from 3.7 +/- 1.8-4.8 +/- 1.9 ml/dl, p < 0.05) at the bypass flow of 10-30 ml/kg per minute in the normal ventilation experiment and at 5 hours after VVECLS initiation in the one lung ventilation experiment (varied from 10.6 +/- 1.6-11.7 +/- 1.8 ml/dl). The blood gas analysis of systemic circulation with balloon inflation revealed that the values of PaO2 (varied from 83.8 +/- 11.4-96.9 +/- 23.4 mm Hg) and PaCO2 (37.7 +/- 9.2-40.4 +/- 11.8 mm Hg) were higher and lower, respectively, compared with balloon deflation. In particular, PaO2 level was significantly higher than that of the preECLS value at the bypass flow of 20-40 ml/kg per minute (varied from 83.8 +/- 11.4-96.9 +/- 23.4 mm Hg, p < 0.05). In the one lung ventilation experiment, systemic PaO2 and PaCO2 levels at balloon inflation were higher and lower, respectively, compared with balloon deflation during VVECLS for 6 hours. At balloon inflation, the value of PaO2 at 6 hours after VVECLS initiation was significantly higher than that at balloon deflation. A newly designed DBDL catheter for ECLS vascular access successfully reduced recirculation and maintained oxygen delivery to systemic circulation during VVECLS. These results suggest that a high bypass flow may not be necessarily required in terms of oxygen delivery to systemic circulation when the DBDL catheter was used as an ECLS vascular access. PMID- 14524570 TI - Treadmill exercise in calves implanted with the AbioCor replacement heart. AB - The aim of this study was to evaluate the function of the AbioCor artificial heart implanted in calves that underwent treadmill exercise testing and to investigate the cardiovascular and metabolic responses during exercise. Six calves were implanted with the AbioCor. One week after surgery, exercise training was undertaken and animals were put on a treadmill at a speed of 0.5-1.0 mph, with the AbioCor operating at the maximum heart rate of 150 beats per minute. During exercise, the left and right atrial pressures were significantly increased (p < 0.05). Oxygen consumption was significantly elevated (p < 0.05). Lactate levels did not significantly increase (p > 0.05). The index of metabolic adequacy remained higher than 2.0, indicating that there was no substantial transition to anaerobic metabolism during exercise. There was an increased extraction of oxygen during exercise to meet the increased energy demands. Our study demonstrated that animals implanted with the AbioCor heart were able to maintain aerobic metabolism during low levels of exercise testing. PMID- 14524571 TI - In vivo performance and biocompatibility of the MagScrew ventricular assist device. AB - Currently available ventricular assist devices (VADs) have limitations in long term durability and blood compatibility. We evaluated a prototype of a pulsatile MagScrew VAD for in vivo hemodynamic performance and biocompatibility. The device is composed of an actuator, blood pump housing, diaphragm, pusher plate, and bioprosthetic valves. Its protein-coated ("biolized") blood-contacting surface inhibits clot formation. Forces between moving parts of the actuator are transmitted magnetically, eliminating a primary source of friction and wear. The pump fills passively and is highly preload sensitive. The device was implanted into three calves for 90, 10, and 57 days, respectively. No anticoagulants were given postoperatively. The device functioned without technical problems during the entire course of each experiment, with mean device flow ranging between 5.4 and 9.0 L/min. Autopsy of the first two calves revealed no sign of embolization and clean blood-contacting surfaces of the devices. The third experiment was complicated by a prosthetic valve endocarditis with infectious embolization, and a few small depositions were found in the pump. In conclusion, the MagScrew VAD has demonstrated a high level of performance and biocompatibility in three calves studied for 10-90 days. Vigorous development is in progress to bring this device to preclinical readiness and thus provide surgeons with the VAD of choice for permanent implantation. PMID- 14524572 TI - Hydrodynamic and static performance evaluation of the moving-actuator type biventricular assist device, AnyHeart. AB - This study evaluated the hydrodynamic characteristics and efficiency of the moving-actuator type implantable biventricular assist device (BVAD), AnyHeart. A blood analog made of 52% glycerin and 48% water was used to simulate the density and viscosity of blood. The maximum pump flow was 9 L/min with 28.8 watts of power input, and the maximum electrical-to-hydraulic power conversion efficiency was approximately 11% at a pump flow of 3.5 L/min. The pump was able to generate 4 L/min output against 100 mm Hg afterload with less than 9 watts of power input. In addition to the overall system efficiency, the inner subpart power conversion efficiency was also evaluated. The system was subdivided according to system mechanism into three major parts: motor part, actuator part, and blood sac part. In normal working conditions (4 L/min, 100 mm Hg) with the AnyHeart, the total system efficiency was 8%, with subpart efficiencies of 50%, 85%, and 19% for motor part, actuator part, and blood sac part, respectively. The pump performance assessed in the in vitro Donovan-type mock circulation loop test was acceptable as a BVAD in terms of flow and pressure. PMID- 14524573 TI - Biventricular support with the Jarvik 2000 axial flow pump: a feasibility study. AB - Patients with congestive heart failure who are supported with a left ventricular assist device (LVAD) may experience right ventricular dysfunction or failure that requires support with a right ventricular assist device (RVAD). To determine the feasibility of using a clinically available axial flow ventricular assist device as an RVAD, we implanted Jarvik 2000 pumps in the left ventricle and right atrium of two Corriente crossbred calves (approximately 100 kg each) by way of a left thoracotomy and then analyzed the hemodynamic effects in the mechanically fibrillated heart at various LVAD and RVAD speeds. Right atrial implantation of the device required no modification of either the device or the surgical technique used for left ventricular implantation. Satisfactory biventricular support was achieved during fibrillation as evidenced by an increase in mean aortic pressure from 34 mm Hg with the pumps off to 78 mm Hg with the pumps generating a flow rate of 4.8 L/min. These results indicate that the Jarvik 2000 pump, which can provide chronic circulatory support and can be powered by external batteries, is a feasible option for right ventricular support after LVAD implantation and is capable of completely supporting the circulation in patients with global heart failure. PMID- 14524574 TI - Reversibility of medically unresponsive pulmonary hypertension with nesiritide in a cardiac transplant recipient. AB - Pulmonary hypertension unresponsive to medical therapy is considered by most centers to be a contraindication for orthotopic cardiac transplantation. This article describes a patient with unresponsive severe pulmonary hypertension who, despite a combination of maximal doses of dobutamine, milrinone, and nitroprusside infusion, responded to nesiritide (Natrecor) infusion with improvement in pulmonary hemodynamics. The patient was considered a high risk for transplantation because of significant pulmonary hypertension in spite of maximum oral therapy and continuous intravenous milrinone. Severe irreversible pulmonary hypertension persisted with a combination of dobutamine, milrinone, and nitroprusside, with pulmonary artery pressure (PA) of 88/44 mm Hg, a transpulmonary gradient (TPG) of 27, and pulmonary vascular resistance (PVR) of 5.79 Wood units. Upon addition of nesiritide, within 24 hours, there was a sustained decrease in PA to 47/30, TPG of 15, and PVR of 3.75 Wood units. The patient underwent successful left ventricular assist device placement soon after nesiritide infusion demonstrated reversibility of pulmonary hypertension. He subsequently underwent uneventful orthotopic cardiac transplantation and has done well with normal right heart pressures. This case illustrates that addition of nesiritide to standard therapy can reverse significant unresponsive pulmonary hypertension and make a patient eligible for left ventricular assist device and orthotopic cardiac transplantation. PMID- 14524575 TI - Percutaneous endoscopic gastrostomy tube in a patient with a left ventricular assist device. AB - We report a successful PEG tube insertion in a patient with an LVAD. Adequate prolonged nutritional support was provided. PEG placement in patients with an implantable or paracorporeal LVAD may be possible but present additional concerns. We believe that PEG placement should be cpnsidered in patients with an LVAD with swallowing difficulties or who are debilitated are not meeting their caloric requirements. PMID- 14524576 TI - Effects of continuous flow left ventricular assist device support on skin tissue microcirculation and aortic hemodynamics. PMID- 14524577 TI - Clinical and pathological features of children with Henoch-Schoenlein purpura nephritis: risk factors associated with poor prognosis. AB - OBJECTIVE: To clarify the risk factors related to prognosis in patients with Henoch-Schoenlein purpura nephritis (HSPN), we investigated the cases with HSPN on long-term observation. METHODS: We enrolled 114 patients who had been diagnosed with HSPN from 1974-1997. These patients were divided into 2 groups based upon features at last follow-up. One group, designated "favorable", consisted of 69 patients with normal urine and 25 patients with minor urinary abnormalities, and the second group, designated "unfavorable", consisted of 15 patients with active renal disease and 5 patients with renal failure. The clinical features, laboratory data and pathological findings were investigated in 2 groups. RESULTS: Nephrotic syndrome, decreased factor XIII activity, hypertension and renal failure at onset were more frequent in "unfavorable" than in "favorable". The rate of glomeruli with crescents, macrophage infiltrations, tubulointerstitial changes and acute exacerbation in "unfavorable" were higher than those in "favorable". There were 5 cases with renal insufficiency, and renal survival rate was 95.6% for over 15 years. CONCLUSIONS: These results suggest that the above mentioned risk factors play an important role in prognosis of the patients with active renal disease and renal failure. PMID- 14524578 TI - Association between blood indoxyl sulfate and carbonyl stress marker in hemodialysis patients. AB - BACKGROUND: Indoxyl sulfate (IS) is a protein-bound solute, which is progressively retained in blood according to the decline of renal function. However, clinical relevance of excess IS in hemodialysis (HD) patients remains unknown. PATIENTS AND METHODS: We measured serum IS and clinical parameters including pentosidine, carboxymethyllysine (CML) and homocysteine levels in 55 HD patients (age: 67 +/- 2 years, time on HD: 67 +/- 11 months, male/female = 30/25), and examined the relationship between IS and these data. RESULTS: Serum IS was markedly increased in HD patients (80.49 +/- 6.17 microg/ml) compared to normal range (< 1.9 microg/ml). IS was significantly and positively correlated with time on HD (p < 0.01), blood urea nitrogen (p < 0.01), beta2-microglobulin (p < 0.03), and protein catabolic rate (PCR) (p < 0.01). The patients with increased IS needed a higher erythropoietin dosage. Blood IS was positively correlated with pentosidine (r = 0.505, p < 0.01) and CML (r = 0.275, p < 0.05). In contrast, blood IS was not associated with nutritional and inflammatory parameters. A stepwise multiple regression analysis revealed that time on HD, PCR and pentosidine were significant determinants of IS. CONCLUSIONS: Serum IS was related to time on HD and dietary protein intake. Accumulated IS may be associated with enhanced carbonyl stress in chronic HD patients. PMID- 14524579 TI - Neither folic nor folinic acid normalize homocysteine levels in hemodialysis patients. AB - AIMS: A greater decrease in total homocystein (tHcy) has been reported in patients on hemodialysis (HD) following the administration of reduced forms of folic acid (FA), however, the effect of the administration of moderated doses of oral levofolinic acid has not been compared with that of FA. We decided to perform a study to evaluate the therapeutic effectiveness of oral levofolinic acid, the pharmacologically active form of folinic acid in our population of HD patients already on treatment with oral FA and vitamin B6. MATERIAL AND METHODS: We undertook a prospective study in HD patients who had been receiving oral supplements of both FA 5 mg every 48 hours and vitamin B6 40 mg every 7 days during at least 6 months, with a 17% initial decrease of tHcy levels. Patients matched for age, sex and time on HD were assigned to 1 of 2 groups: Those in group A continued to receive their previous supplements while in group B, FA was substituted by calcium levofolinate 5 mg given orally every 48 hours. The following parameters were measured at baseline and at month 6: urea kinetic model and concentrations of plasma albumin, C-reactive protein, folate, vitamin B12, pyridoxal phosphate and tHcy. RESULTS: Group A: 30 patients aged 63.4 (57.9, 68.9) years, with a time on HD of 23.4 (15.8, 30.8) months, group B: 32 age matched patients 66.2 (62.1, 70.3) years old, with a time on HD of 23.8 (16.7, 30.9) months. No differences were found either in folate levels (72.7 (47.9, 97.5) vs. 71.9 (44.0. 99.9) ng/ml), tHcy (23.5 (21.1, 25.9) vs. 23.3 (20.8, 25.8) micromol/l), or any other study variables. In the 2 groups a significant reduction in both residual renal function (RRF) and vitamin B12 levels was observed after supplementation, but no changes in tHcy values, folate levels or any of the other parameters were found. The prevalence of hyperhomocysteinemia in group A was 93.3% at study start and 100% at month 6, in group B the corresponding values were 93.8% and 96.9%. After 6 months, multiple regression analysis showed that tHcy levels were not influenced by the type of treatment (p = 0.543). CONCLUSIONS: After 6 months of calcium-levofolinate supplementation tHcy levels did not decrease and were similar to those in patients given the same dose of FA. PMID- 14524580 TI - Simultaneous hemodialysis during coronary angiography fails to prevent radiocontrast-induced nephropathy in chronic renal failure. AB - BACKGROUND: Radiocontrast medium- (RM) associated nephrotoxicity continues to be a common cause of acute renal failure and may lead in patients with pre-existing chronic renal insufficiency even to end-stage renal failure requiring chronic dialysis. Since extracorporeal removal of RM after RM administration has been shown to be effective but does not prevent radiocontrast-induced nephropathy, the effect of a simultaneous dialysis during RM administration on renal function is not clear. METHODS: In a prospective, randomized and controlled trial, we studied the effect of a 4-hour online dialysis during RM (iomeprol) application in patients with advanced chronic renal failure (serum creatinine > or = 3 mg/dl) undergoing coronary angiography. All patients received hydration with saline before and after standardized coronary angiography and were randomized to receive a simultaneous high-flux hemodialysis (7 patients, HD group) or to control group (10 patients). 24-hour creatinine clearance (CrCl) was measured in all patients before, 1 week and 8 weeks after coronary angiography. The clinical follow-up comprised 8 weeks after RM application. RM plasma levels were measured in both groups 15, 30, 60 minutes, 2, 4, 12, 24, 48 and 72 hours after application by high-pressure liquid chromatography. RESULTS: At baseline, CrCl (19 +/- 10 vs 17 +/- 7 ml/min), percentage of diabetics (57 vs 70%) and dose of RM (77 +/- 27 vs 86 +/- 21 ml) were similar in both groups. Pharmacokinetics: Total clearance of iomeprol was significantly higher (54 +/- 15 vs 20 +/- 12 ml/min, p < 0.001) and the area under curve (AUC) was significantly lower (23 +/- 10 g x h/l vs 94 +/- 57 g x h/l, p < 0.001) in the HD group compared to control group. RM peak plasma levels 15 min after application were not different in both groups (3.0 +/- 1.1 vs 4.2 +/- 1.7 mmol/l, NS), however, significantly lower 60 min (1.6 +/- 0.4 vs 3.7 +/- 1.5 mmol/l, p < 0.01) and 240 min (0.7 +/- 0.3 vs 2.3 +/- 0.7 p < 0.001) after angiography. CLINICAL RESULTS: CrCl showed no difference 1 week (24 +/- 11 vs 19 +/- 9 ml/min, ns) and 8 weeks (24 +/- 5 vs 20 +/- 9 ml/min, NS) after angiography from baseline or between the groups. In each group, 2 patients developed end-stage renal disease and requested permanent dialysis during the 8 week follow-up. CONCLUSION: Simultaneous dialysis reduces AUC of iomeprol significantly, however, does not influence plasma peak concentration after angiography. Renal function and incidence of end-stage renal failure were not influenced by online-dialysis. PMID- 14524581 TI - Relationship between different body size indicators and hernia development in CAPD patients. AB - AIMS: A small body size may increase the risk for hernia development in patients on continuous ambulatory peritoneal dialysis (CAPD). The present study investigates whether there is a relationship between body size and hernia development in CAPD patients. MATERIAL AND METHODS: The records of 78 patients on CAPD were reviewed retrospectively. Body mass index (BMI), body surface area (BSA) and total body water (TBW) were calculated in all patients. Correlations between different body size indicators (BMI, BSA and TBW) and hernia development were assessed using analysis of covariance in which we adjusted for sex. RESULTS: A total of 14 patients (17.9%) with no physical evidence of hernia before catheter insertion developed hernias. Body size was significantly lower in CAPD patients with hernias than those without hernias when adjusted for sex. CONCLUSIONS: We conclude that patients with small body size tend to have an increased risk for hernia development. A simple estimation of patients' height, weight, body surface area and total body water would be helpful to predict development of hernias or other complications related to increased intraperitoneal pressure in CAPD patients. PMID- 14524582 TI - Lupus-like nephritis in an HIV-positive patient: report of a case and review of the literature. AB - The most common manifestation of HIV/AIDS in the kidney is the collapsing variant of focal segmental glomerular sclerosis, HIV-associated nephropathy (HIVAN). Other forms of renal disease in HIV-infected patients include mesangial proliferative glomerulonephritis (GN), membranoproliferative GN, IgA nephropathy, minimal change disease and proliferative immune-complex GN. We present the case of a 42-year-old Caucasian male with HIV infection, treatment associated peripheral neuropathy, nephrotic syndrome and progressive renal failure. The initial and subsequent kidney biopsies showed diffuse proliferative glomerulonephritis resembling diffuse proliferative (WHO class IV) lupus nephritis. There was no clinical or serological evidence of systemic lupus erythematosus (SLE). Proteinuria improved with ACE-inhibitors, and renal function remained relatively stable while receiving highly active antiretroviral therapy (HAART). A precipitous decline in renal function to end-stage renal disease followed a brief period of withdrawal from potent antiretroviral therapy during which the viral load rebounded. Considering previously reported cases, it appears that lupus-like nephritis is a rare but well-defined pattern of immune-complex induced renal injury seen in HIV-infected patients. It appears to be markedly responsive to HAART. PMID- 14524583 TI - Alport syndrome and benign familial hematuria (thin basement membrane disease) in two brothers of a family with hematuria. AB - Alport syndrome (AS) and benign familial hematuria (BFH) are inherited disorders of the glomerular basement membrane, which are sometimes difficult to differentiate at the early stage without type IV collagen staining of the renal basement membrane. Previous studies have indicated that mutation of type IV collagen alpha4 gene may be responsible for both BFH and AS. We report here a Japanese family with consanguinity, in which autosomal-recessive AS and BFH were separately identified in two brothers on the basis of findings of electron microscopy and type IV collagen chain staining of the renal biopsy specimens. Their parents, being first cousins, paternal uncle and grandmothers were found to have hematuria. Our observations suggest that BFH patients were heterozygous carriers of autosomal-recessive AS. PMID- 14524584 TI - Interstitial nephritis associated with perianeurysmal retroperitoneal fibrosis. AB - Renal failure in retroperitoneal fibrosis is usually secondary to ureteric obstruction. We report a patient whose renal failure did not improve following relief of obstruction. Renal biopsy showed an interstitial nephritis, and function improved with steroid therapy. We suggest that renal biopsy should be considered in cases of renal failure associated with retroperitoneal fibrosis that do not respond to the relief of ureteric obstruction. PMID- 14524585 TI - Recurrent severe anion gap metabolic acidosis secondary to episodic ethylene glycol intoxication. AB - Acute ethylene glycol toxicity and its attendant metabolic derangement is a well described clinical entity. Recurrent severe anion gap metabolic acidosis consequent to episodic ingestion of ethylene glycol has not been previously reported. We present a patient who developed severe anion gap metabolic acidosis with no osmolar gap and hypokalemia, consequent to episodic ethylene glycol ingestion. Modest artifactual elevation of the serum lactic acid level and rapid response to intravenous bicarbonate infusion may serve as diagnostic clues. Consideration of these aberrant features should be included in the clinical assessment of severe anion gap metabolic acidosis. PMID- 14524586 TI - Worsening of hypertension in a pregnant woman with renal arteriovenous malformation: a successful superselective embolization after delivery. AB - A 30-year-old female presented with uncontrolled hypertension due to arteriovenous malformation in the upper third of the right kidney, which worsened during pregnancy. The arteriovenous malformation was detected by color-coded Doppler sonography, confirmed by angiography, and the fistula was sealed by superselective arterial embolization with metallic coils. Superselective embolization is the most effective and safe treatment for this rare and complex pathology. PMID- 14524587 TI - A case of anaphylactoid shock induced by the BS polysulfone hemodialyzer but not by the F8-HPS polysulfone hemodialyzer. AB - A 57-year-old woman was admitted because of severe bradycardia and hypotension caused by an anti-arrhythmic agent and beta-blocker. For 19 months before admission, she had been undergoing hemodialysis with an F8-HPS polysulfone membrane hemodialyzer without any complications. In 2 dialysis sessions after admission, when a BS polysulfone membrane was used, she experienced anaphylactoid shock with severe hypotension leading to syncope, dyspnea and vomiting, just after the start of hemodialysis. After the anaphylactoid shock, her dialyzer membrane was changed to a cellulose triacetate membrane and she did not suffer from such attacks. This case indicates that severe anaphylactoid shock may be caused by a biocompatible dialyzer membrane and that the reactions of patients to each polysulfone membrane may differ among polysulfone membranes made by different manufacturers. PMID- 14524588 TI - Clopidogrel-induced membranous nephropathy. PMID- 14524589 TI - Gene polymorphism in the promoter region of lipopolysaccharide receptor CD14 and dyslipidemia in hemodialysis patients. PMID- 14524590 TI - Partial exchange transfusion improved renal function and proteinuria in a patient with cyanotic heart disease nephropathy. PMID- 14524591 TI - A case of encircling penile condylomata acuminata after renal transplantation. PMID- 14524592 TI - Anticoagulation for mechanical valve protheses during pregnancy. PMID- 14524593 TI - Determining prognosis of the individual patient. PMID- 14524594 TI - Oral platelet glycoprotein IIb/IIIa receptor inhibitors--part II. AB - Although the hypothesis of benefit from prolonged oral IIb/IIIa inhibition was appealing, the large Phase III trials have uniformly shown there was no improvement in outcome. In addition, there was an increased mortality seen in patients treated with the oral IIb/IIIa inhibitor. This latter finding is not adequately explained, but is likely a multifactorial problem of this strategy of platelet inhibition. The trials found that, even with no improvement in efficacy, there was increased bleeding, meaning that for chronic therapy with IIb/IIIa inhibition there does not appear to be a therapeutic window. Accordingly, chronic oral IIb/IIIa inhibition appears to have been well tested but has not worked. Fortunately, there are several other oral antiplatelet agents available that have shown beneficial results, including clopidogrel. In addition, other newer classes of antiplatelet agents are in earlier stages of development. Thus, agents targeted more "upstream" in platelet activation pathways may offer a more tolerable and efficacious approach to long-term antiplatelet therapy. PMID- 14524596 TI - Images in cardiology: Partial anomalous pulmonary venous drainage in adulthood. PMID- 14524595 TI - Hyperhomocysteinemia in patients with heart failure referred for cardiac transplantation: preliminary observations. AB - BACKGROUND: Hyperhomocysteinemia is becoming recognized as a risk factor for cardiovascular disease, yet there are limited data on the prevalence of hyperhomocysteinemia in patients with heart failure. HYPOTHESIS: The purpose of this study was to examine the prevalence of hyperhomocysteinemia in patients with severe heart failure and to correlate serum homocysteine levels with factors that may affect homocysteine metabolism. METHODS: Serum homocysteine levels were measured at the time of cardiac transplant evaluation in 89 consecutive patients with severe heart failure. Homocysteine levels for patients with ischemic cardiomyopathy (ICM) were compared with levels obtained in patients with nonischemic cardiomyopathy (NICM), and homocysteine levels were correlated with demographic and hemodynamic parameters as well as functional status. RESULTS: The mean plasma homocysteine level was increased (14.3 +/- 5.3 micromol/l, normal <9.0 micromol/l) and was equivalent between patients with ICM versus NICM (14.7 +/- 5.8 micromol/l vs. 13.8 +/- 4.5 micromol/l, p = 0.44). Elevated homocysteine levels were seen in a large proportion (89%) of patients and were equally common to patients with NICM (94%) and ICM (85%). Serum homocysteine levels correlated with serum creatinine (r = 0.51, p < 0.001), with a history of diabetes (p = 0.028), and with a history of peripheral vascular disease (p = 0.045). Only 6% of patients were receiving folic acid therapy at the time of transplant referral. CONCLUSION: Hyperhomocysteinemia is common in patients with severe heart failure, and plasma homocysteine levels are uniformly elevated regardless of the etiology of heart failure. Elevated plasma homocysteine levels are likely a consequence of heart failure-related renal insufficiency. PMID- 14524598 TI - Images in cardiology: Vintage Vineberg! PMID- 14524597 TI - Predictors of cause-specific hospital readmission in patients with heart failure. AB - BACKGROUND: Repeated hospital readmissions are frequent and increasing over time in patients with heart failure (HF). The predictors for readmission in patients with HF are not completely understood. HYPOTHESIS: The study was undertaken to investigate the time course of readmission by specific cause in patients with HF, and to examine the independent effects of HF etiology and left ventricular (LV) function on cause-specific readmissions. METHODS: A retrospective cohort of 493 consecutive patients with HF was followed for readmission for 16.5 +/- 12.3 months. Ischemic etiology of HF was defined as history of myocardial infarction (MI), coronary artery bypass graft (CABG), percutaneous transluminal coronary angioplasty (PTCA), or > or = 70% coronary stenosis. Left ventricular function was assessed echocardiographically. Cause-specific readmissions were classified as HF, cardiovascular disease (CVD) other than HF, and other non-CVD. RESULTS: The annual readmission rate was 56.6%. Median time to readmission was 91 days, with 18.3% patients readmitted within 1 month after discharge. Ischemic etiology independently predicted all-cause readmission: Cox hazard ratio (95% confidence interval): 1.40 (1.11-1.79). This relationship was significant in women (1.83 [1.31-2.55]), but not in men (1.15 [0.82-1.62]), while readmissions were equally frequent in both genders. Similarly, ischemic etiology significantly predicted readmission for CVD in women (4.18 [2.14-8.19]), but not in men (1.49 [0.83 2.67]). However, LV dysfunction independently predicted readmission for recurrent HF (2.44 [1.46-4.08]), while ischemic etiology was not predictive in either gender. CONCLUSIONS: Readmissions for recurrent HF comprise only one-third of total hospital readmissions in patients with HF. Ischemic etiology is a significant predictor of readmission, and most of this effect is mediated through a four-fold increased risk of readmission for CVD other than HF in women. Readmission for recurrent HF is predicted by LV dysfunction but not by ischemic etiology. Patients with HF can be accurately risk stratified for cause-specific readmission with available clinical data. PMID- 14524599 TI - Reversal of heart failure remodeling: is it maintained? AB - BACKGROUND: Reversal of heart failure remodeling has been observed with intensive heart failure therapy. HYPOTHESIS: We hypothesized that reversal of heart failure remodeling may not be sustained in long-term follow-up. METHODS: Sixty-one sequential patients with heart failure and left ventricular ejection fraction < or = 35%, who improved their ejection fraction by > or = 10% over baseline at follow-up, were prospectively followed and retrospectively analyzed. Each patient underwent echocardiography at baseline and biannually thereafter. RESULTS: In all patients, left ventricular ejection fraction increased from 18 +/- 7% to 42 +/- 12% on uptitrated medical therapy. At follow-up over 20 +/- 8 (+/- standard deviation) months, this improvement was sustained in 38 patients ("Improved"). A relapse in remodeling occurred in the remaining 23 patients ("Relapsed"), with ejection fraction falling to 24 +/- 7%. For Improved and Relapsed patients, baseline echocardiographic and clinical parameters were equivalent. However, Improved patients tended to be younger, with shorter heart failure duration. Improved patients had more effective improvement in ejection fraction than Relapsed patients (45 +/- 13% vs. 36 +/- 8%, p = 0.005), with greater reductions in chamber size and mitral regurgitation. CONCLUSION: Reversal of heart failure remodeling may be sustained in only two-thirds of patients at long-term follow up. In contrast to Relapsed patients, Improved patients tended to be younger, with shorter heart failure duration and a more complete recovery of left ventricular systolic function. PMID- 14524600 TI - Effects of acute angiotensin-converting enzyme inhibition on diastolic ventricular interaction in the dilated heart. AB - BACKGROUND: The normal and dilated heart behaves as a single functional unit during preload reduction: volume unloading in the setting of diastolic ventricular interaction allows for increased left ventricular (LV) filling. HYPOTHESIS: We hypothesized that reduction of venous return induced by a physiologic stimulus (tilting) or by acute angiotensin-converting enzyme (ACE) inhibitors in dilated heart is likely to have a marked and similar effect on ventricular chamber geometry and filling. This study was designed to assess how the normal and dilated heart adapts to preload reduction. METHODS: Twenty normal subjects and 20 patients with moderate heart failure due to dilated cardiomyopathy were studied with two-dimensional and Doppler echocardiography in supine position (B) and after 40 degrees of head-up tilting (T). The following day, patients repeated supine (C) and tilting test (TC) after administration of captopril (25 mg s.l.). Right ventricular (RV) and LV dimensions, LV geometry, and tricuspid, mitral, and pulmonary venous flow patterns were recorded at each step of the study. RESULTS: In the two groups, T was associated with reduction of RV area and LV volumes; C and TC produced a similar effect on RV and LV. Changes in LV septal-lateral diameter and anterior-posterior diameter were different at each step of the study: during T (both groups) and after C and TC, the septallateral diameter increased slightly while the anterior-posterior diameter decreased. During T, mitral and tricuspid peak flow velocities decreased, peak late velocities were unchanged, and the deceleration time of mitral flow increased; the systolic forward flow of pulmonary venous flow decreased, the diastolic forward flow did not change, and the difference in duration between reverse pulmonary flow and mitral peak late flow decreased: C and CT induced similar changes. CONCLUSION: Preload reduction induced by tilting or by ACE inhibitors induces profound and similar effects on LV and RV dimensions, LV geometry, and biventricular filling. Reduction of RV dimension is associated with adaptation of LV geometry and decrease of LV diastolic pressure, which facilitates LV filling and pulmonary venous drainage: ACE inhibition associated with tilting exerts an additional effect on these changes. These data confirm the role of ventricular interaction in modulating LV filling in heart failure. PMID- 14524602 TI - The effects of aging on atrial endocardial electrograms in patients with paroxysmal atrial fibrillation. AB - BACKGROUND: The prevalence of atrial fibrillation (AF) has been reported to increase with advancing age. Histologic studies in AF have demonstrated that the percentage of fibrosis and degenerative changes in the atrial muscle increase significantly with age. HYPOTHESIS: This study was undertaken to assess the influence of advancing age on atrial endocardial electrograms recorded during sinus rhythm in patients with paroxysmal atrial fibrillation (PAF), which had not been assessed previously. METHODS: Right atrial endocardial catheter mapping during sinus rhythm was performed in 111 patients with PAF to evaluate the influence of advancing age on atrial endocardial electrograms. The bipolar electrograms were recorded at 12 sites in the right atrium, and an abnormal atrial electrogram was defined as lasting > or = 100 ms, and/or showing eight or more fragmented deflections. RESULTS: In all, 1,332 right atrial endocardial electrograms were assessed and measured quantitatively. The number of abnormal atrial electrograms in patients with PAF showed a significantly positive correlation with age (r = 0.34; p < 0.0005). Patients aged > 60 years had a significantly greater mean number of abnormal electrograms (2.58 +/- 2.05) than those aged < 60 years (1.43 +/- 2.03; p < 0.004). The longest duration (r = 0.35; p < 0.0005) and the maximal number of fragmented deflections (r = 0.29; p < 0.005) of atrial electrograms among the 12 right atrial sites also showed a significantly positive correlation with age. CONCLUSIONS: Aging alters the electrophysiologic properties of the atrial muscle in patients with PAF. Elderly patients have a significantly greater abnormality of atrial endocardial electrograms than do younger ones. There is a progressive increment in the extension of altered atrial muscle with advancing age in patients with PAF. PMID- 14524601 TI - Detection of patients with hypertrophic cardiomyopathy at risk for paroxysmal atrial fibrillation during sinus rhythm by P-wave dispersion. AB - BACKGROUND: Paroxysmal atrial fibrillation (PAF) in hypertrophic cardiomyopathy (HCM) is associated with poor prognosis. Previous studies have shown good correlation between P-wave dispersion (Pd) and occurrence of PAF. However, Pd in patients with HCM for predicting PAF has not been studied. HYPOTHESIS: The aim of the study was to determine whether Pd could identify patients with HCM who are likely to suffer from PAF. METHODS: Twenty-two patients with HCM with a history of PAF (Group 1) and 26 patients with HCM without a history of PAF (Group 2) were studied. Maximum (Pmax) and minimum (Pmin) P-wave durations, as well as P-wave dispersion (Pd = Pmax - Pmin) were calculated from 12-lead surface electrocardiograms (ECG). RESULTS: P-wave dispersion was significantly different between the groups (Group 1: 55 +/- 6 ms vs. Group 2: 37 +/- 8 ms; p<0.001), while Pmax (Group 1: 134 +/- 11 ms vs. Group 2: 128 +/- 13 ms; p = 0.06) and Pmin (Group 1: 78 +/- 9 ms vs. Group 2: 81 +/- 7 ms; p = 0.07) was not significantly different. Patients with a history of PAF had higher left atrial diameter than the patients without PAF (Group 1: 52 +/- 8 mm vs. Group 2: 48 +/- 10 mm; p = 0.02). A cut-off value of 46 ms for Pd had a sensitivity of 76% and a specificity of 82% in discriminating between patients with and without PAF. CONCLUSION: This study suggests that P-wave dispersion could identify patients with HCM who are likely to develop PAF. PMID- 14524603 TI - Right atrial sensing for VDD pacing through a persistent left superior vena cava. AB - This report describes a pacing lead insertion through a persistent left superior vena cava in the VDD pacing mode, with a large retrograde lead course allowing the optimal positioning of the sensing electrodes in the right atrium. PMID- 14524604 TI - Images in cardiology: Vieussens' ring. PMID- 14524605 TI - The heart-lung machine was invented twice--the first time by Max von Frey. PMID- 14524606 TI - Past and present concepts in flow cytometry: a European perspective. AB - The development of flow cytometric instrumentation, methods and research concepts in Europe has been a continuous driving force for the general scientific advancement in this area over the years. This review addresses early European concepts of continuing interest with regard to instrumentation, data analysis, clinical and eperimental DNA analysis, cell function and microbiology at their worldwide first appearence while flow cytometric immunology and immunophenotyping will be covered separately. Flow cytometry represents an efficient approach to the enormous complexity of molecular cell architecture and cell function by the analysis of apparent molecular cell phenotypes in heterogeneous cell samples. The present merger of flow and image cytometry into the method independent cytomics discipline increases the potential of cell analysis very significantly. It opens the way for predictive medicine as well as for predictive cytopathology and predictive cytology in everyday clinical and medical practice. Current progress is driven by joint advances in molecular fluorescence technologies and instrument development. This complements the analysis of genome sequence information in an efficient way. PMID- 14524607 TI - Setting up and calibration of a flow cytometer for multicolor immunophenotyping. PMID- 14524608 TI - Low level leucocyte counting (LLLC) in blood products. PMID- 14524609 TI - Flow cytometric screening for HLA-B27 on peripheral blood lymphocytes. PMID- 14524611 TI - Immunophenotypic characterization of CD34(pos) cells. PMID- 14524610 TI - Single platform enumeration of viable CD34(pos) cells. PMID- 14524612 TI - Appendix 1: auto-standardization and compensation for CD34 analysis (Beckman Coulter XL Flow Cytometer - System II Software). PMID- 14524613 TI - Appendix 2: ammonium chloride lysing solution. PMID- 14524614 TI - Flow cytometric enumeration of Class I HLA-restricted, peptide-specific CD8+ T lymphocytes using tetramer technology and single-platform absolute T-cell counting. PMID- 14524615 TI - The role of non-chromosomal histones in the host defense system. AB - Histone H1 is located at the inter-nucleosome and more correctly at both ends of the double-stranded DNA that protrude from the nucleosome unit. It has long been recognized to be localized only inside the nuclei as a constituent for packaging nucleosome into chromatin. Thus, it could be hardly believed that detatched or solubilized histone H1 plays the role of a host defense molecule. Given the old reports on histone-like basic proteins that show bacteriostatic functions, I herein chose some recent related articles and tried review them. Recent advances in research on the cell death mechanism makes it possible to understand that programmed cell death, (i.e. apoptosis) could serve as a good source of soluble histones. Some forms of them are highly probable to be bacteriostatic. PMID- 14524616 TI - Molecular analysis of fecal microbiota in elderly individuals using 16S rDNA library and T-RFLP. AB - Fecal microbiota in six elderly individuals were characterized by the 16S rDNA libraries and terminal restriction fragment length polymorphism (T-RFLP) analysis. Random clones of 16S rRNA gene sequences were isolated after PCR amplification with universal primer sets from total genomic DNA extracted from feces of three elderly individuals. These clones were partially sequenced (about 500 bp). T-RFLP analysis was performed using 16S rDNA amplified from six subjects. The lengths of the terminal restriction fragment (T-RF) were analyzed after digestion by HhaI and MspI. Among 240 clones obtained, approximately 46% belonged to 27 known species. About 54% of the other clones were 56 novel "phylotypes" (at least 98% homology of clone sequence). These libraries included 83 species or phylotypes. In addition, about 13% (30 phylotypes) of these phylotypes were newly discovered in these libraries. A large number of species that are not yet known exist in the feces of elderly individuals. 16S rDNA libraries and T-RFLP analysis revealed that the majority of bacteria were Bacteroides and relatives, Clostridium rRNA cluster IV, IX, Clostridium rRNA subcluster XIVa, and "Gammaproteobacteria". The proportion of Clostridium rRNA subcluster XIVa was lower than in healthy adults. In addition, although Ruminococcus obeum and its closely related phylotypes were detected in high frequency in healthy young subjects, hardly any were detected in our elderly individuals. "Gammaproteobacteria" were detected at high frequency. PMID- 14524618 TI - Phylogenetic characterization of Orientia tsutsugamushi isolated in Taiwan according to the sequence homologies of 56-kDa type-specific antigen genes. AB - Fourteen strains of Orientia tsutsugamushi isolated in Taiwan were characterized by sequencing 56-kDa type-specific antigen genes and patterns of restriction fragment length polymorphism (RFLP) predicted by a computer program. The strains showed high varieties in sequence homologies and were classified to 10 types by predicted patterns of RFLP. Furthermore, all Taiwan strains were not identical in typing with strains analyzed previously. These results suggest that there are various types of O. tsutsugamushi in Taiwan that are different from those distributed in other countries. PMID- 14524617 TI - Cloning and expression of Bartonella henselae sucB gene encoding an immunogenic dihydrolipoamide succinyltransferase homologous protein. AB - Immunoscreening of a ZAP genomic library of Bartonella henselae strain Houston-1 expressed in Escherichia coli resulted in the isolation of a clone containing 3.5 kb BamHI genomic DNA fragment. This 3.5 kb DNA fragment was found to contain a sequence of a gene encoding a protein with significant homology to the dihydrolipoamide succinyltransferase of Brucella melitensis (sucB). Subsequent cloning and DNA sequence analysis revealed that the deduced amino acid sequence from the cloned gene showed 66.5% identity to SucB protein of B. melitensis, and 43.4 and 47.2% identities to those of Coxiella burnetii and E. coli, respectively. The gene was expressed as a His-Nus A-tagged fusion protein. The recombinant SucB protein (rSucB) was shown to be an immunoreactive protein of about 115 kDa by Western blot analysis with sera from B. henselae-immunized mice. Therefore the rSucB may be a candidate antigen for a specific serological diagnosis of B. henselae infection. PMID- 14524619 TI - A quantitative in vitro assay method for detecting biological activity of endotoxin using prostaglandin E2 induction in rabbit peripheral blood. AB - The pyrogen test and the endotoxin test (the LAL test) have been playing crucial roles in detecting endotoxin in parenteral drugs. The current test methods, however, have disadvantages such as requiring a large number of animals or an inadequacy in evaluation of in vivo endotoxin activity. We attempted to establish a new assay method that can overcome the shortcomings of the current methods. We standardized the in vitro assay method by the use of prostaglandin E2 (PGE2) induction from peripheral blood of rabbits for detecting endotoxin activity. A linear dose-response regression was attained from approximately 0.15 to 5 endotoxin units/ml of Japanese national reference standard endotoxin by the in vitro assay. The assay showed a fine correlation with the pyrogen test but not with the LAL test, when endotoxins from various bacterial sources were tested. The in vitro assay was also shown to have the capability of detecting a synergistic effect of endotoxin and parenteral drugs. The in vitro PGE2 induction test using rabbit blood was, therefore, suggested to be the appropriate test method for guaranteeing the same level of safety of parenteral drugs as the pyrogen test does. PMID- 14524620 TI - Distribution of human rotaviruses, especially G9 strains, in Japan from 1996 to 2000. AB - A 4-year (1996-2000) survey of rotavirus infection involving 2,218 diarrheal fecal specimens of children collected from five regions of Japan was conducted. A total of 642 (28.9%) specimens were found to be rotavirus positive. A changed prevalence pattern of rotavirus G serotype was found with an increase of G9 and G2 and a decrease of G1, although G1 remained the prevailing serotype. Serotype G9 was unexpectedly determined to be the prevailing serotype in Sapporo (62.5%) and Tokyo (52.9%) in 1998-1999, and in Saga (78.4%) in 1999-2000. G9 strains isolated from 1998-1999 belonged to the P[8]-NSP4-Wa-group with long RNA pattern, while, G9 strains isolated from 1999-2000 belonged to three groups, the P[8]-NSP4 Wa-group with long RNA pattern, the P[4]-NSP4-KUN-group with short RNA pattern and a mixed-type group (P[4]/P[8]-NSP4-KUN/Wa-group with long RNA pattern). Both sequence and immunological analysis of VP7 revealed that the G9 strains from 1999 2000 were much more closely related to the G9 strains isolated worldwide in the 1990s, including G9 strains found in Thailand in 1997. However, the G9 strains from 1998-1999 were distinct from these and more closely related to the G9 prototype strains F45, AU32 and WI61 discovered in Japan and the US in the 1980s. Thus the G9 strains isolated in 1998-1999 had progenitors common to the G9 prototype strains, while the strains isolated in 1999-2000 did not directly evolve from them but were related to global G9 strains that have emerged in recent years. These data supported our previous report that G9 rotavirus might exist as two or more subtypes with diverse RNA patterns, P-genotype and NSP4 genogroup combinations (Y.M. Zhou et al., J. Med. Virol. 65: 619-628, 2001) and suggested that G9 rotavirus prevalent in Japan during two successive years belonged to different subtypes. The nucleotide sequences presented in this paper were submitted to DDBJ, EMBL and GenBank nucleotide sequence databases. The accession numbers are: 00-Ad2863VP7 (AB091746), 00-OS2986VP7 (AB091747), 00 SG2509VP7 (AB091748), 00-SG2518VP7 (AB091749), 00-SG2541 (AB091750), 00-SG2864 (AB091751), 00-SP2737VP7 (AB091752), 99-SP1542VP7 (AB091753), 99-SP1904VP7 (AB091754), 99-TK2082VP7 (AB091755) and 99-TK2091VP7 (AB091756). PMID- 14524621 TI - Hepatitis C virus nonstructural protein NS3 binds to Sm-D1, a small nuclear ribonucleoprotein associated with autoimmune disease. AB - Hepatitis C virus (HCV) causes a persistent infection, chronic hepatitis, and leads to hepatocellular carcinoma. Nonstructural protein 3 (NS3) of HCV possesses protease, nucleoside triphosphatase, and helicase activities. Using the yeast two hybrid assay, we identified Sm-D1, a host protein that binds to NS3. Sm-D1 is a component of small nuclear ribonucleoprotein (snRNP) complexes which are associated with autoimmune disease. Sm-D1 has Gly-Arg (GR) repeats at the C terminus, which contains dimethylarginine modified by post-translational modification and may constitute an immunoreactive determinant. Deletion mutants revealed that the C-terminal region of Sm-D1 containing the GR repeats was the binding region for NS3, and the expression feature of Sm-D1 was affected by co expression of NS3. Immunostaining assay demonstrated that NS3 was also present in the nucleus of cells overexpressing Sm-D1, although it was usually found in cytoplasm. The localization of NS3 could change following interaction with Sm-D1 and affect the function of Sm-D1. PMID- 14524622 TI - Improved recovery of rabies virus from cloned cDNA using a vaccinia virus-free reverse genetics system. AB - To improve efficiency of recovery of rabies virus from cloned cDNA, we established a BHK cell clone that stably expresses T7 RNA polymerase, which we named BHK/T7-9. We also constructed new helper plasmids for expression of nucleoprotein and RNA polymerase of the RC-HL strain using the pTM1 plasmid vector, which makes the T7 RNA polymerase-transcripts from the plasmid cap independent for translation. After co-transfection of these helper plasmids and the previously constructed full-length genome plasmid of the RC-HL strain to BHK/T7-9 cells, recombinant rabies virus was efficiently recovered from the cloned cDNA. PMID- 14524623 TI - "The silver lining". PMID- 14524624 TI - An ethicist's commentary on the case of the client seeking antibiotic in feed. PMID- 14524625 TI - Antimicrobial susceptibility of hazard analysis critical control point Escherichia coli isolates from federally inspected beef processing plants in Alberta, Saskatchewan, and Ontario. AB - A survey to estimate the prevalence of antimicrobial resistance in Escherichia coli was conducted in 7 Canadian federally inspected processing plants during 2001. Escherichia coli isolates were recovered during routine hazard analysis critical control point sampling from beef carcasses and trim and subsequently tested for their antimicrobial susceptibility by using susceptibility panels. Of the 2653 isolates analyzed, 68% were sensitive to all 18 antimicrobials tested. For 14 of the 18 antimicrobials evaluated, the percentage of resistant isolates was < or = 1. Twenty-five percent of the isolates were resistant to tetracycline, 9% to sulfamethoxazole, 7% to streptomycin, and 3% to ampicillin. Multiple resistance was found in 12% of the isolates, with 7% showing resistance to 2 antimicrobials, 2% to 3 antimicrobials, 2% to 4 antimicrobials, and 1% to 5 or more antimicrobials. Forty-five different antimicrobial resistance patterns were observed. The reasons for the development of the antimicrobial resistance were not investigated in this study. This study was useful as a pilot to help to develop a national antimicrobial resistance surveillance program in Canada. This study indicates that laboratory standardization is possible for consistent results across the country and that the indicator organism, E. coli, is fairly easy to obtain for surveillance but Salmonella are not, due to their low prevalence in beef. PMID- 14524626 TI - Computed tomographic appearance of epidural empyema in a dog. AB - The computed tomographic (CT) appearance of epidural empyema in a 5.5-month-old, intact male boxer dog is described. Epidural empyema was diagnosed by means of CT, surgery, and bacterial culture. The imaging and pathogenesis of epidural abscesses and the pitfalls of differentiating caudal fossa lesions from severe cranial cervical lesions are discussed. PMID- 14524627 TI - Diagnosis of trichomoniasis in 'virgin' bulls by culture and polymerase chain reaction. AB - The diagnostic test for Tritrichomonas foetus in bulls is microscopic examination of cultured preputial samples. Trichomonads other than T. foetus can be present in a preputial sample. Both a staining technique and a polymerase chain reaction assay were useful in differentiating between T. foetus and another trichomonad observed in samples from virgin bulls. PMID- 14524628 TI - Retrospective analysis of etiologic agents associated with respiratory diseases in pigs. AB - Twenty-eight hundred and seventy-two cases of respiratory disease in pigs were analyzed for their etiologic agents. Two or more pathogens were detected from 88.2% of the cases, indicating that porcine reproductive and respiratory syndrome virus (PRRSV) or swine influenza virus (SIV) combined with other bacterial agents was a common cause for porcine respiratory diseases in the mid-western USA. PMID- 14524629 TI - Congenital nutritional muscular dystrophy in a beef calf. AB - A 13-hour-old Aberdeen-Angus was involuntarily recumbent since birth. Congenital nutritional muscular dystrophy was suspected based on clinical findings, increased serum creatine kinase, and decreased serum vitamin E and selenium levels. Recovery followed after supportive therapy and parenteral vitamin E and selenium. Reports of this disease in newborn calves are unusual. PMID- 14524630 TI - Transmural rectal intestinal evisceration associated with parturition in a primiparous mare. AB - A 6-year-old, primiparous standardbred mare was presented with a history of intestinal protrusion from the rectum shortly after parturition. A subacute grade IV rectal tear was diagnosed. The unusual nature of the tear led to speculation of a possible owner palpation accident. A grave prognosis was given and the mare was euthanized. PMID- 14524631 TI - Malignant giant cell tumor of soft parts in a mare. AB - Two subcutaneous masses were removed from the elbow of a mare. Histologically they were composed of islands of polygonal to plump spindlelioid cells with large nuclei, coarsely stippled chromatin, and eosinophilic cytoplasm. Findings were diagnostic for a malignant giant cell tumor of soft parts, a rare tumor with a fair prognosis. PMID- 14524632 TI - Cholangiohepatitis and pancreatitis secondary to severe gastroduodenal ulceration in a foal. AB - A 2-month-old foal was presented with clinical signs of colic. Gastroduodenal ulceration was suspected. A poor response to medical treatment and signs of gastroduodenal obstruction led to celiotomy and an attempted bypass procedure. The foal was euthanized and postmortem examination revealed gastric ulceration, segmental duodenal stenosis, and severe chronic cholangiohepatitis and pancreatitis. PMID- 14524633 TI - Can we assess welfare? PMID- 14524634 TI - What is mentorship? PMID- 14524635 TI - Acute bilateral ocular proptoses and mild right pigmentary keratitis. PMID- 14524637 TI - What's new in lipid management? AB - Statins are the principal agents for managing hyperlipidemia because of their ability to reduce low-density lipoprotein cholesterol (LDL) and triglyceride levels, increase high-density lipoprotein cholesterol (HDL) levels slightly, and decrease clinical events. Nevertheless, not all patients can tolerate the statins or achieve their treatment goals with these drugs. In addition, available statins have only modest effects on HDL, and clinical events still occur despite the best current lipid-lowering agents. Thus, new lipid-lowering agents are required to improve clinical outcomes. These recently or soon to be released agents are stanol esters, which can be incorporated into spreads and other foods and reduce LDL levels up to 15%; colesevelam, a new bile acid sequestrant with a higher potency, fewer adverse effects, and less drug interactions than those of older agents in this class; rosuvastatin, a new statin that reduces LDL levels and increases HDL levels at least more effectively than atorvastatin; and a new class of agents that inhibit cholesterol absorption and are likely to be used in combination with statins. Low-density lipoprotein cholesterol apheresis is another, nonpharmacologic option for patients with severe hyperlipidemia despite maximal medical therapy. PMID- 14524636 TI - Update on the National Cholesterol Education Program Adult Treatment Panel III guidelines: getting to goal. AB - Considerable data on the pathophysiology, epidemiology, and treatment of dyslipidemia-induced coronary heart disease (CHD) have accumulated in recent years. These data have been assessed and incorporated into the guidelines of the National Cholesterol Education Program Expert Panel on the Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel [ATP] III). A major focus of the new guidelines is the assessment of the near-term (i.e., 10-yr) risk of experiencing a CHD event and matching the intensity of treatment to this risk. Patients with diabetes and those with a greater than 20% 10-year risk of experiencing a CHD event have been elevated to the risk level of CHD equivalent. The ATP III guidelines also modify several lipid and lipoprotein classifications. A low-density lipoprotein cholesterol (LDL) level below 100 mg/dl is now considered optimum for all individuals. In addition, high-density lipoprotein cholesterol (HDL) and triglyceride cutoff points have been modified to reflect more accurately the risk associated with abnormalities in these lipoproteins. As with the previous guidelines, the primary target of therapy remains LDL. Therapeutic lifestyle changes consisting of diet, weight reduction, and increased physical activity should be included in all treatment regimens. Based on their potent LDL-lowering properties and their proven ability to decrease mortality in a variety of patient populations, statins are generally the first choice for pharmacologic therapy. A secondary target of therapy includes non-HDL goals for patients with high triglyceride levels and the metabolic syndrome, which is characterized by abdominal obesity, elevated triglyceride levels, low HDL levels, and insulin resistance. Management of these secondary targets includes weight reduction and increased physical activity, and treatment of the lipid and nonlipid risk factors. Overall, ATP III represents an aggressive approach to treating dyslipidemia, greatly extending the number of individuals who qualify for treatment. PMID- 14524638 TI - Role of the pharmacist in establishing lipid intervention programs. AB - Despite the availability of the National Cholesterol Education Program Adult Treatment Panel (ATP) guidelines for the management of hyperlipidemia since 1988, most patients do not achieve their target low-density lipoprotein cholesterol (LDL) goals. With the publication of the most recent guidelines (ATP III), which contain more aggressive treatment recommendations, the cholesterol treatment gap is likely to widen further. Factors responsible for patients not receiving adequate treatment include a lack of focus on asymptomatic diseases, time and reimbursement constraints, inadequate training, a reluctance to prescribe aggressive treatment regimens, and poor communication among health care professionals. Results of several studies evaluating intervention programs indicate that pharmacists can play a key role in improving cholesterol management whether in lipid clinics, community pharmacies, or hospitals. In these intervention programs, pharmacists provided a wide range of functions that included reviewing the medical history, monitoring laboratory values, selecting lipid-lowering therapies, and educating patients regarding drug therapies and the importance of compliance. These interventions produced significant improvements in lipid parameters and in the number of patients who achieved LDL treatment goals. Most important, these interventions were associated with decreases in clinical events. Pharmacist intervention also was highly cost-effective and time efficient. These results suggest that pharmacists are in a unique position and possess the requisite skills to improve the treatment of patients with hyperlipidemia. PMID- 14524639 TI - Osmolality and osmolarity: narrowing the terminology gap. PMID- 14524641 TI - Propylene glycol-associated renal toxicity from lorazepam infusion. AB - OBJECTIVES: Using data from patients who developed elevations in serum creatinine concentrations while receiving continuous-infusion lorazepam, we sought to determine the correlations between the magnitude of serum creatinine concentration rise and each of the following variables: serum propylene glycol level, cumulative lorazepam dose, and duration of lorazepam administration. An additional objective was to identify clinical markers for propylene glycol toxicity. DESIGN: Retrospective chart review. SETTING: Medical-surgical intensive care unit and burn unit at a university hospital. PATIENTS: Eight patients who developed elevations in serum creatinine concentrations while receiving continuous-infusion lorazepam (range 2-28 mg/hr). MEASUREMENTS AND MAIN RESULTS: The mean cumulative dose of lorazepam was 4305 mg (range 1200-10,920 mg), and the mean propylene glycol level determined at the time of peak serum creatinine concentration was 1103 microg/ml (range 186-3450 microg/ml). Serum creatinine concentrations increased in all eight patients during lorazepam infusion and decreased in seven within 3 days after stopping infusion. A weak-to-moderate correlation existed between the magnitude of the rise in serum creatinine concentration and propylene glycol level (r=0.53). A weak-to-moderate correlation also was identified between cumulative lorazepam dose and magnitude of serum creatinine concentration rise (r=0.43), and a strong-to-moderate correlation was found between duration of lorazepam infusion and magnitude of serum creatinine concentration rise (r=0.60). Propylene glycol levels were strongly correlated with both serum osmolality and osmol gap. CONCLUSION: The patients' increased serum creatinine concentrations are likely to have resulted from exposure to propylene glycol as a result of lorazepam infusion. Serum osmolality and osmol gap may be useful markers for propylene glycol toxicity. PMID- 14524640 TI - Comparing the bispectral index and suppression ratio with burst suppression of the electroencephalogram during pentobarbital infusions in adult intensive care patients. AB - OBJECTIVE: The bispectral index (BIS), a processed variable derived from the raw electroencephalogram (EEG) used to guide sedation in the intensive care unit (ICU), has not been tested during barbiturate therapy for elevated intracranial pressure. We determined the BIS and suppression ratio (SR) values during traditional burst monitoring of the raw EEG during pentobarbital infusions. DESIGN: Prospective, observational cohort study. SETTING: A 42-bed multidisciplinary ICU in a tertiary care medical center. PATIENTS: Twelve consecutive patients with elevated intracranial pressure treated with pentobarbital infusions. INTERVENTION: All patients were monitored continuously with the Aspect Medical Systems A-1050 bedside EEG monitor using a bilateral referential montage. Pentobarbital doses were titrated based on the raw EEG to attain a burst-suppression pattern with a goal of 3-5 bursts/minute. Drug dosage, intracranial pressure, cerebral perfusion pressure values, EEG bursts/minute, BIS version 3.2, and SR were recorded daily. MEASUREMENTS AND MAIN RESULTS: The 12 patients were monitored for 62 patient-days. Mean +/- SD age was 32 +/- 15 years, seven (58%) patients were male, mean Acute Physiology and Chronic Heath Evaluation II score was 17.0 +/- 5.0, and hospital mortality was 42%. The mean pentobarbital infusion rate was 124 +/- 49 mg/hour or 2.3 +/- 1.3 mg/kg/hour, and mean pentobarbital serum concentration was 29.7 +/- 13 microg/ml. The mean BIS value was 18 +/- 14, mean SR 56% +/- 36%; BIS correlated well with SR (r=-0.99, p<0.001). For patient-days with a burst-suppression pattern, BIS 3.2 (r=0.90, p<0.001) and SR (r=-0.89, p<0.001) strongly correlated with the number of bursts/minute. The mean BIS value corresponding to 3-5 bursts/minute was 15 (95% confidence interval [CI] 10-20); SR value was 71 (95% CI 61-80). CONCLUSION: The Aspect A-1050 applied to patients and monitored by nurses and physicians works well as a bedside EEG monitor, providing a raw EEG signal to titrate barbiturate therapy. The continuous data trend and real-time digital output for the BIS and SR quantify the degree of EEG suppression well and may prove helpful in facilitating titration of barbiturate infusions. PMID- 14524642 TI - Outcomes of dual-protease inhibitor salvage therapy in human immunodeficiency virus-infected patients referred to a telephone consultation service. AB - OBJECTIVE: To determine the clinical outcomes of dual-protease inhibitor salvage therapy in heavily experienced patients after their providers consulted the National HIV Telephone Consultation Service (Warmline, 800-933-3413). DESIGN: Observational survey study. SETTING: Consultation service for United States health care providers. PATIENTS: Thirty-one human immunodeficiency virus (HIV) infected patients who had received previous treatment with at least two protease inhibitor--or nonnucleoside reverse transcriptase inhibitor--based regimens and had a viral load of at least 1000 copies/ml. INTERVENTION: Patients whose providers consulted Warmline regarding antiretroviral salvage regimens were identified through a review of telephone calls from January-July 2001. Virologic and immunologic outcomes were determined for patients who had received a regimen containing either ritonavir and indinavir or ritonavir and amprenavir. MEASUREMENTS AND MAIN RESULTS: Primary outcomes were percentages of patients with a viral load less than 500 copies/ml at 3 and 6 months and changes in log10 viral load and CD4+ cell count at 3 and 6 months compared with baseline values. By using intent-to-treat analysis, a viral load less than 500 copies/ml was achieved in 35% of the 31 patients at 3 months and in 32% of them at 6 months. By using as treated analysis, this outcome was achieved in 48% of 23 patients who continued therapy at 3 months and in 59% of 17 patients who continued therapy at 6 months. At 3 months (23 patients) and 6 months (17 patients), respectively, changes in viral load were -1.7 log10 copies/ml (p<0.001) and -1.4 log10 copies/ml (p<0.001), and changes in CD4+ cell count were +99 cells/mm3 (p<0.001) and +95 mm3 (p=0.012), compared with baseline. CONCLUSION: Significant improvements in virologic and immunologic markers occurred in patients heavily experienced with antiretroviral therapy after starting dual-protease inhibitor treatment regimens. Salvage therapy guided by Warmline consultation appears to be beneficial for this patient population with limited treatment options. PMID- 14524643 TI - Essential tremor: diagnosis and treatment. AB - Essential tremor is a common movement disorder in adults that interferes with the performance of functional and social activities. Differentiation of essential tremor from other tremor syndromes is important in order to provide appropriate patient education and therapy. The mainstays of pharmacotherapy are propranolol and primidone; however, in selected patients, agents such as alcohol, benzodiazepines, botulinum toxin, and gabapentin may provide symptomatic benefits. Advances in surgical interventions, such as stereotactic thalamotomy and thalamic deep brain stimulation, offer patients an alternative treatment modality when pharmacotherapy is inadequate. A treatment algorithm is provided to guide clinicians in the management of patients with essential tremor. PMID- 14524644 TI - Pancreatitis associated with atypical antipsychotics: from the Food and Drug Administration's MedWatch surveillance system and published reports. AB - STUDY OBJECTIVE: To investigate the relative numbers and clinical characteristics of pancreatitis in patients treated with the atypical antipsychotic agents, clozapine, olanzapine, and risperidone, versus the conventional neuroleptic, haloperidol. DESIGN: Pharmacovigilance study of pooled, spontaneously reported adverse events. SETTING: Government-affiliated drug evaluation center. PATIENTS: One hundred ninety-two patients who developed pancreatitis during treatment with one or more antipsychotic agents. INTERVENTION: Patients were identified with the Food and Drug Administration's MedWatch surveillance program and a MEDLINE search. MEASUREMENTS AND MAIN RESULTS: Most cases of pancreatitis occurred within 6 months after the start of therapy with one or more antipsychotic agents. Of the reports of pancreatitis occurring in conjunction with these drugs, 40%, 33%, 16%, and 12% were in patients receiving treatment with clozapine, olanzapine, risperidone, and haloperidol, respectively. In 50% of the patients receiving haloperidol, an atypical antipsychotic was listed as a concomitant drug. Valproate was administered concomitantly in 23% of patients. Hyperglycemia and acidosis, although uncommon, developed with all the drugs except haloperidol. Twenty-two patients died. In contrast to patients who developed pancreatitis while receiving an atypical antipsychotic, those who developed the disease while receiving haloperidol were women and tended to be older. CONCLUSION: The number of reports involving the three atypical antipsychotic agents and the relative paucity of reports involving haloperidol, despite its more extensive patient exposure, suggest that atypical antipsychotics may precipitate pancreatitis. However, the risk may not be the same with all agents; pancreatitis was reported most frequently with clozapine, followed by olanzapine, and then risperidone. The temporal relationship of the onset of pancreatitis with the start of drug therapy further supports a cause-and-effect relationship. PMID- 14524645 TI - Treatment of premenstrual dysphoric disorder with selective serotonin reuptake inhibitors. AB - Premenstrual dysphoric disorder (PMDD) is considered a severe form of premenstrual syndrome. Symptoms of PMDD occur during the last week of the luteal phase of the menstrual cycle and usually abate at the onset of menses. About 3-8% of all menstruating women experience PMDD, which can lead to significant functional impairment. Several randomized, controlled trials have assessed the efficacy of selective serotonin reuptake inhibitors (SSRIs) in the treatment of PMDD. The SSRIs were found to significantly improve symptoms, particularly psychological or behavioral symptoms, during the luteal phase in women with PMDD. Also, SSRIs were found to improve the quality of life in women with PMDD. Headache, fatigue, insomnia, and anxiety were often reported as adverse effects. A decrease in libido or sexual dysfunction also was reported. In recent studies, intermittent SSRI therapy was found to be effective treatment for PMDD and allows a woman to take the drug for only 14 days each month. Intermittent SSRI therapy should be recommended before continuous daily dosing of SSRIs in the treatment of PMDD. PMID- 14524646 TI - Pharmacologic blockade of the renin-angiotensin system: vascular benefits beyond commonly understood pharmacologic actions. AB - Angiotensin-converting enzyme (ACE) inhibitors and angiotensin II receptor blockers (ARBs) are recognized primarily for their use in hypertension, in heart failure, and after myocardial infarction. New evidence, particularly with ACE inhibitors, has shown their ability to reduce acute coronary events associated with atherosclerosis in patients without a history of the aforementioned cardiac conditions. This is likely due to inhibitory effects on the renin-angiotensin system--a system that adversely influences fibrinolytic balance, vascular endothelial function, and vascular inflammation, all key components of atherosclerotic progression and adverse coronary outcomes. Results of various studies suggest favorable effects of ACE inhibitors and ARBs on markers of these components, including effects on plasminogen activator inhibitor-1, endothelin-1, and nitric oxide by ACE inhibitors, and effects on vascular cell adhesion molecule-1 and C-reactive protein by ARBs. Although early evidence suggests that ACE inhibitors may provide a greater beneficial effect on some of these markers compared with ARBs, and that certain ACE inhibitors may provide greater vascular benefits than others, further investigation is required to verify such findings. Overall, understanding the distinct coronary vascular benefits of these agents will emphasize the importance of using them, particularly ACE inhibitors, to improve outcomes in patients with coronary atherosclerotic disease. PMID- 14524647 TI - A primer for developing pharmacist-managed clinics in the outpatient setting. AB - Pharmacist-managed clinics exist in many practice settings, such as physician offices, hospital-based outpatient clinics, and pharmacies. This article is intended to serve as a primer for pharmacists who wish to establish clinical pharmacy services in these settings. Recommendations for beginning this process are delineated. Information on building initial support, developing a needs assessment analysis, selecting the focus of new clinical services, developing a detailed clinic proposal, and marketing new services is provided. In a comprehensive review of how to develop a clinic proposal, the purpose, background information, proposed clinic structure, implications, assessment, billing issues, financial summary, and resources requested are discussed. Opportunities for financial compensation in various practice settings, such as the physician outpatient clinic, hospital-based outpatient clinic, and community pharmacy, are also reviewed. PMID- 14524648 TI - Impact of atypical coverage for patients with community-acquired pneumonia managed on the medical ward: results from the United States Community-Acquired Pneumonia Project. AB - STUDY OBJECTIVE: As current guidelines for treatment of community-acquired pneumonia (CAP) recommend empiric antimicrobial coverage for atypical pathogens, we evaluated the need for atypical coverage by examining length of hospital stay (LOS) and mortality in patients with CAP who were managed on the medical ward. METHODS: Medical records of patients with CAP admitted from January 1, 1997 December 31, 2001, from 176 United States nonteaching community hospitals were reviewed. Patients were divided into one of three mutually exclusive groups on the basis of intravenous antimicrobials received on days 1 or 2 of hospital stay: ceftriaxone monotherapy, ceftriaxone plus a macrolide, or levofloxacin. Variables evaluated for their ability to predict outcome were patient age, year of hospital admission, geographic region, preadmission setting, preadmission antimicrobial treatment, timing of antimicrobial administration, comorbid disease, and duration of intravenous antimicrobial treatment. The impact of initial antimicrobial regimen on LOS and mortality was evaluated in regression models while controlling for significant predictors of outcome. RESULTS: Of 8975 patients evaluated, 2453 met the inclusion criteria. Significant differences were noted among patients who received ceftriaxone (932 patients), ceftriaxone plus a macrolide (872), and levofloxacin (649) with respect to mean +/- SD age (72 +/- 16, 67 +/- 18, and 70 +/- 17 yrs, respectively; p<0.0001), admission from a nursing home (21%, 11%, and 15%, respectively; p<0.0001), and duration of intravenous antimicrobial treatment (4.4 +/- 2.7, 4.0 +/- 2.6, and 3.6 +/- 2.5 days, respectively; p<0.0001). The LOS predictors were age, geographic region, coexisting heart failure, and duration of intravenous antimicrobial therapy. Mortality predictors were age, admission from a nursing home, coexisting heart failure, and coexisting cancer. After controlling for these predictors of outcome, no significant differences were noted among the three groups for LOS (5.5 +/- 3.5, 4.8 +/- 2.9, and 4.8 +/- 2.9 days, respectively; p=0.2791) or mortality (3.1%, 2.0%, and 2.6%, respectively; p=0.8461). CONCLUSION: Initial coverage for atypical pathogens does not affect LOS or mortality among patients with CAP managed on the medical ward. PMID- 14524649 TI - Impact of a collaborative care model on depression in a primary care setting: a randomized controlled trial. AB - To measure the effects of a collaborative care model that emphasized the role of clinical pharmacists in providing drug therapy management and treatment follow-up to patients with depression, we conducted a randomized controlled trial at a staff model health maintenance organization. We compared the outcomes of subjects treated in this collaborative care model (75 patients, intervention group) with subjects receiving usual care (50 patients, control group). After 6 months, the intervention group demonstrated a significantly higher drug adherence rate than that of the control group (67% vs 48%, odds ratio 2.17, 95% confidence interval 1.04-4.51, p=0.038). Patient satisfaction was significantly greater among members randomly assigned to pharmacists' services than among controls, and provider satisfaction surveys revealed high approval rates as well. Changes in resource utilization were favorable for the intervention group, but differences from the control group did not achieve statistical significance. Clinical improvement was noted in both groups, but the difference was not significant. Clinical pharmacists had a favorable effect on multiple aspects of patient care. Future studies of this model in other health care settings appear warranted. PMID- 14524650 TI - Superwarfarin and glass ingestion with prolonged coagulopathy requiring high-dose vitamin K1 therapy. AB - A 23-year-old man was brought to the emergency department after eating four boxes of brodifacoum-containing rodenticide over a 4-day interval and pieces from approximately two bottles of glass over the previous 2 weeks. He was asymptomatic but his prothrombin time was markedly elevated with an international normalized ratio (INR) of 37.8. A plain abdominal film showed diffuse radiopaque foreign bodies, presumably glass, in the large and distal small intestines. Treatment for ingested glass consisted of stool softeners and bulk-forming laxatives. The patient developed mild gingival bleeding and received fresh frozen plasma (FFP) infusions and vitamin K1 orally. At a vitamin K1 dosage of 300 mg/day, the INR corrected to less than 2.0 and the patient was discharged taking that dosage. He returned 26 days later with hematuria and flank pain, and his INR was 189. He was administered FFP and packed red blood cells, and his vitamin K1 dosage was increased to 800 mg/day; his INR returned to baseline. Compliance with taking the vitamin K1, which required ingestion of 60-160 tablets/day, was a serious problem, requiring numerous follow-up calls and visits to the patient at home and work. At 5-month follow he was doing well. Compliance with large daily doses of vitamin K1 for treatment of "superwarfarin" ingestion may be poor because of the duration of treatment and large number of pills required. A more concentrated formulation may be advantageous for management of patients with brodifacoum poisoning. PMID- 14524651 TI - Acute tubular necrosis associated with propylene glycol from concomitant administration of intravenous lorazepam and trimethoprim-sulfamethoxazole. AB - A 46-year-old morbidly obese man was admitted to the medical intensive care unit with respiratory failure. He required pressure-control ventilation and high levels of sedation with continuous-infusion lorazepam. He developed Stenotrophomonas maltophilia pneumonia; treatment included scheduled intravenous trimethoprim-sulfamethoxazole. Each of these drugs contain several hundred milligrams/milliliter of propylene glycol. On day 17 of his hospital course, 3 days after starting the trimethoprim-sulfamethoxazole, the patient developed acute renal failure consistent with acute tubular necrosis. Propylene glycol toxicity was suspected; therefore, all drugs containing propylene glycol were discontinued, and laboratory data were collected. A marked osmol gap, metabolic acidosis, and renal toxicity were attributed to both continuous and large intermittent doses of intravenous propylene glycol. Particular attention should be paid to the total amount of propylene glycol provided to patients from administered drugs. Patients in the intensive care setting who require high doses of intravenous lorazepam for sedation, as well as antimicrobial therapy with trimethoprim-sulfamethoxazole for treatment of either Stenotrophomonas maltophilia or Pneumocystis carinii pneumonia, may be at increased risk for propylene glycol toxicity and should be monitored closely. PMID- 14524652 TI - Vancomycin-induced thrombocytopenia: a case proven with rechallenge. AB - In a rare case of vancomycin-induced thrombocytopenia, a 50-year-old man with culture-negative subacute bacterial endocarditis underwent mitral valve replacement surgery and was treated with vancomycin. His platelet count dropped from 346 x 10(3)/mm3 to 13 x 10(3)/mm3 on postoperative day 4, and a differential diagnosis of heparin- versus drug-induced thrombocytopenia was considered. Antiheparin antibodies were detected in the patient's serum on day 5. He showed no signs of bleeding. His platelet count remained below 5 x 10(3)/mm3 despite two platelet transfusions on day 5. A hemorrhagic pericardial effusion with tamponade developed, requiring drainage. A trial with intravenous immunoglobulin led to fever and chills, and the infusion was not completed. Vancomycin was changed to clindamycin on day 9, and methylprednisolone therapy was started on day 11. On day 12, the patient's clinical condition improved, and his platelet count increased from 3 x 10(3)/mm3 to 32 x 10(3)/mm3 with no bleeding. On day 18, his platelet count was 424 x 10(3)/mm3, and he was scheduled for discharge with vancomycin therapy for a total of 6 weeks. He received a single dose of intravenous vancomycin 1 g at the hospital; his platelet count dropped to 160 x 10(3)/mm3 1 hour after the infusion and to 58 x 10(3)/mm3 12 hours later. Vancomycin was discontinued and clindamycin and prednisone were restarted. On day 20, the patient's platelet count increased to 105 x 10(3)/mm3 and he was discharged with warfarin, prednisone, and clindamycin therapy. We suspect that our patient's thrombocytopenia was due to vancomycin. PMID- 14524653 TI - Acute renal failure secondary to angiotensin II receptor blockade in a patient with bilateral renal artery stenosis. AB - A 54-year-old man with diabetes mellitus, peripheral vascular disease, and hypertension was admitted to the hospital for an acute exacerbation of chronic heart failure. Therapy with intravenous furosemide and oral losartan 100 mg twice/day was begun. Ten days later, the patient's blood urea nitrogen and serum creatinine levels rose and peaked at 110 and 6.0 mg/dl, respectively. His serum potassium level increased to 5.7 mg/dl, urine output dropped to 400 ml over 24 hours, and mental status changes occurred. Magnetic resonance angiography revealed bilateral renal artery stenosis. After losartan was discontinued and hemodialysis was performed for 3 consecutive days, the patient's renal function returned to his baseline level. Reports in the medical literature reinforce the importance of recognizing that angiotensin-converting enzyme inhibitors should be used with caution in patients with bilateral renal artery stenosis. However, the literature is not as definitive about using of angiotensin II receptor blockers (ARBs) in these patients. Our patient's experience suggests that ARBs should be used with caution in patients with bilateral renal artery stenosis. Clinicians should be aware that renal failure might occur when using ARBs in these patients. PMID- 14524654 TI - From club drug to orphan drug: sodium oxybate (Xyrem) for the treatment of cataplexy. AB - Narcolepsy, a rare disease with a prevalence of 0.05% in the general population, affects an estimated 140,000 patients in the United States. Patients have been able to lead fuller personal and professional lives since the Food and Drug Administration approved sodium oxybate (Xyrem) in 2002 for treatment of cataplexy in patients with narcolepsy. Previously, gamma-hydroxybutyrate (GHB), the active ingredient of sodium oxybate, had been a substance of abuse, most notoriously as a date-rape drug. Public Law 106-172, the date-rape prohibition act enacted in 2000, was modified to allow the drug to be legally administered for medical purposes. Because of the apprehension regarding the risk of possible drug diversion after the approval of sodium oxybate and concerns about safety, the Xyrem Risk Management Program was created. This program has been successful in satisfying the needs of patients and physicians while ensuring responsible distribution of the drug. PMID- 14524655 TI - Collaborative drug therapy management by pharmacists--2003. AB - Since publication of the initial ACCP position statement on CDTM by pharmacists in 1997, the public, government, and much of the health care community at large have come to better appreciate the growing complexity of providing effective and safe drug therapy in today's health care environment. Increased interest in the issues of cost and quality of drug use is evident in the increasing coverage of the issue in the lay press and professional literature. This represents real progress, as well as real opportunity, for pharmacists. It also heightens the potential for a better understanding of the vital role that pharmacists can play in addressing these concerns. The percentage of patients who take several drugs for chronic diseases will continue to increase. Based on current trends, the number of patients who lack adequate access to care, or who receive either suboptimal, inappropriate, or unnecessarily expensive drug therapy for their acute and chronic diseases, will increase. Even as financial and human resources are increasingly strained within the current health care system, costs will continue to rise unless changes are made. Fortunately, qualified pharmacists are prepared, capable, and willing to help address a significant portion of these challenges. The public, many health care providers, some legislators, and a few insurers now recognize that pharmacists, because of their education and training in drug therapy, are well positioned both to accept additional responsibility for patient care and to provide services that make a real difference in health care quality and outcomes. The health care programs administered by the U.S. Public Health Service, the armed forces, and the Veterans Health Administration, as well as 38 states, now support pharmacist participation in CDTM. Pharmacists, working in an interdisciplinary structure with physicians and other health care providers, have demonstrated that they can improve the effectiveness, efficiency, and safety of drug therapy by providing CDTM. It is time to incorporate this valuable professional skill of the contemporary pharmacist as a core component of the delivery of health care services. PMID- 14524656 TI - Molecular analysis of microbial communities in nitrification and denitrification reactors treating high ammonia leachate. AB - Molecular analysis of microbial populations in two bench-scale nitrification and denitrification reactors fed with high ammonia landfill leachate was conducted in this study by using DGGE, cloning, and FISH techniques in addition to classical efficiency control parameters. Nitrification tank was operated with a computer controlled alkalinity dosing system to supply the alkalinity intermittently as consumed on the basis of on-line pH monitoring. By keeping the pH at 7.0 with this system, 99% nitrification efficiency and rates of about 0.14-0.18 mgNH4+ N/mgVSSday were obtained. Meanwhile, as ammonia oxidizing bacteria Nitrosomonas and Nitrosococcus mobilis-like cells and as nitrite oxidizing bacteria Nitrobacter-related cells were intensively indicated. Moreover, some aerobic denitrifiers as Thauera species were also identified. After the termination of pH adjustment in the preceding anaerobic reactors, nitrification tank was loaded with more biodegradable COD as a result of reduced COD removal in anaerobic reactors. Microbial diversity was immediately affected from this alteration and heterotrophic carbonaceous bacteria and aerobic denitrifiers have dominated. To provide the former high efficiencies, retention time has increased from 24 to 48 h and a second pump dosing HCl was included to the automatic control system. Subsequent to these precautions, numbers of ammonia (Nso190) and nitrite oxidizing bacteria (NIT3) were comparatively increased. In denitrification system, about 98% denitrification efficiencies were obtained at 2000 mg/L NOx-N concentrations if sodium acetate was supplied as carbon source. Meanwhile, with 20 gVSS/l biomass concentration, denitrification rates of about 1.34 mgNOx N/mgVSSday were obtained. All sludge samples have represented similar DGGE patterns and Paraccoccus-related species were identified as dominant denitrifying bacteria. PMID- 14524657 TI - Image analysis as a monitoring tool for activated sludge properties in lab-scale installations. AB - An important step in the battle against filamentous bulking is the development of a monitoring system for activated sludge properties. Therefore, a fully automatic image analysis method for recognizing and characterizing flocs and filaments in activated sludge images has been developed. This procedure has been subsequently used to monitor activated sludge properties in a lab-scale installation. The results of a 100-days experiment indicate that the image information correlates well with the evolution of standard settling properties, in this case the Sludge Volume Index. It is shown that, at the onset of severe filamentous bulking, there is an increase in total filament length on the one hand, and a significant change in floc shape on the other hand. PMID- 14524658 TI - Degradation of organic substances and reactive dye in an immobilized-cell sequencing batch reactor operation on simulated textile wastewater. AB - Textile wastewater generally consists of high organic substances and is strongly colored. Reactive dye has been used extensively in the textile industries. It is water soluble and difficult to remove by chemical coagulation. Removal of organic substances simultaneously with dye can be achieved by a biological process. This study aims to investigate the treatability of the organic substances and reactive dye in immobilized-cell sequencing batch reactors (SBR). Three different supporting medias namely activated carbon, steel slag and plastic were used. The performance of each reactor was compared with a conventional sequencing batch reactor. The simulated textile wastewater containing the reactive azo dye Procion Red H-E7B of a concentration of 40 mg/L and COD 300 mg/L, was fed into the reactors. The supporting media in the SBR system, it will enhance the capability of COD and dye operating of the SBRs consisted of 5 periods; Fill 1.5 h, React (anoxic:oxic) 20 (14:6) h, Settle 1.5 h, Draw 0.5 h and Idle 0.5 h. The results revealed that by adding removal. During a steady state of operation, the COD and dye concentrations of each period were investigated. In addition, the prolonged anoxic period brought about better decolorization efficiency. PMID- 14524659 TI - Microbial population dynamics in an anaerobic CSTR treating a chemical synthesis based pharmaceutical wastewater. AB - Effects of a chemical synthesis based pharmaceutical wastewater on performance of an anaerobic completely stirred tank reactor (CSTR), activity of acetoclastic methanogens and microbial composition were evaluated under various influent compositions. Initially, the CSTR was fed with glucose up to an organic loading rate (OLR) of 6 kg COD/m3 x d corresponding to an F/M ratio of 0.43 with a hydraulic retention time (HRT) of 2.5 days. A COD removal efficiency of 92% and a methane yield of 0.32 m3 CH4/kg COD(removed) were achieved whilst specific methanogenic activity (SMA) was found to be 336mL CH4/gTVS x d. After the CSTR was fed with pre-aerated wastewater diluted by glucose in different dilution ratios of 10% (w/v), 30% (w/v), 70% (w/v), and 100% (w/v) pre-aerated wastewater, gradual decreases in COD removal efficiency to 71%, methane yield to 0.28 m3CH4/kg COD(removed) and SMA to 166 mL CH4/gTVS d occurred whilst volatile fatty acid concentration reached to 1474 mg/L. After the raw wastewater diluted with the pre-aerated wastewater was fed into the CSTR in increasing ratios of 10% (w/v), 30% (w/v), and 60% (w/v), there was a proportional deterioration in performance in terms of COD removal efficiency, methane yield and acetoclastic methanogenic activity. Epifluorescence microscopy of the seed sludge revealed that Methanococcus-like species, short, and medium rods were found to be equally dominant. The short and medium rod species remained equally dominant groups in the CSTR throughout the feeding regime whilst Methanococcus-like species and long rods were found to be in insignificant numbers at the end of the study. Changes in archael diversity were determined using molecular analyses such as polymerase chain reaction (PCR), and denaturent gradient gel electrophoresis (DGGE). Results showed that overall archeal diversity did not change much whereas changes in composition of eubacterial population occurred. PMID- 14524660 TI - Anaerobic biodegradability and treatment of Egyptian domestic sewage. AB - The anaerobic biodegradability of domestic sewage for four Egyptian villages and four Egyptian cities was determined in batch experiments. The results showed that the biodegradability of the Egyptian-villages sewage (73%) was higher than that of the cities (66%). The higher biodegradability of the soluble COD of village sewage (69%) as compared to that of the cities (46%) was the reason for the higher biodegradability of the total COD of the villages sewage. The biodegradability of suspended COD was similar for the sewage of both villages and cities (73-74%). The results of a mathematical-model. developed based on anaerobic digestion model number 1, indicate that at applying a UASB reactor for the treatment of Egyptian villages and cities sewage, an optimum HRT of, respectively. 16 and 8 h is required. At these HRTs, a total COD removal and a conversion to methane of, respectively, 62-70% and 59-64% can be achieved for the sewage of cities and, respectively, 71-77% and 67-69% for the villages sewage. The model results also show that in the treatment of villages sewage in a two step (anaerobic filter + UASB reactor) system a higher total COD removal can be obtained (77-81%) at a short HRT of 10 h (4 + 6 h). However, the excess sludge from the first-step of the two-step system will be less stabilized. PMID- 14524661 TI - Anaerobic treatment of low-strength wastewater in a biofilm reactor. AB - A 3.9-L (70-cm high) upflow anaerobic filter (AF) packed with corrugated plastic rings was employed to evaluate the direct treatment of low-strength wastewater over a long period of operation (32 months at 25.4 degrees C and 2 months at 15.5 degrees C). The AF was fed four complex synthetic-type wastewaters with a low solids content [chemical oxygen demand (COD) 325-403 mg L(-1), soluble COD (SCOD) 86-339 mg L(-1) and total suspended solids (TSS) 65-156 mg L(-1)] at organic loading rates (OLR) ranging from 0.02 to 0.91 kg COD m(-3) d(-1). Operation with filtered municipal wastewater supplemented with a sugar-nutrient based substrate (OLR 0.27-0.91 kg COD m(-3) d(-1)) yielded removal efficiencies in the range of 72-80% for COD and 80-92% for TSS, and use of a completely synthetic substrate resulted in improved effluent quality. Under these test conditions, removal of organic matter and solids occurred within the first 40cm of column height and accumulation within the reactor was not substantial or affected by the length of operation. PMID- 14524662 TI - A dynamic model for anaerobic filter. AB - Anaerobic filters are frequently used to treat medium to high strength wastewater (2000-20000 mg/L COD), but have fewer applications to low strength wastewater (< 1000 mg/L COD). In order to understand the applicability of anaerobic treatment for low strength wastewater, such as domestic sewage, a dynamic mathematical model was developed. This Monod-type kinetic model, which predicts treatment efficiency and gas production, was used to describe literature observations and is offered as a predictive tool. PMID- 14524663 TI - Pharmaceutical wastewater treatment using an anaerobic/aerobic sequencing batch biofilter. AB - The performance of a sequencing batch biofilter integrating anaerobic/aerobic conditions in one tank to treat a pharmaceutical wastewater effluent was studied. A pilot reactor, packed with a porous volcanic stone (puzzolane) was used in the study. The reactor operated as a sequencing batch biofilter, SBB, with reaction times varying for the anaerobic stage from 8 to 24 h and for the aerobic one from 4 to 12 h. The volume of exchange was from 16 to 88%. The pharmaceutical wastewater contained organic chemicals including phenols and o-nitroaniline, a concentration of organic matter that varied from 28,400 to 72,200 mg/L (as total COD), 280 to 605 mg N-NH4/L. and 430 to 650 mg SST/L. In order to acclimatize the microorganisms to the industrial wastewater, the organic load was increased stepwise from 1 to 7.7 kg COD/m3/d. The adequate time was obtained when the removal efficiency of COD reached 80%, or more. Maximal removal loads, associated to high removal efficiencies (95-97% as COD), varied from 4.6 to 5.7 kg COD/m3/d. Under these conditions color removal was 80% as Pt-Co units. Microtox analysis was performed to the wastewater and to the anaerobic and aerobic stages. It was observed that the aerobic stage was the responsible for wastewater detoxification. Results showed that the anaerobic/aerobic SBB was able to treat efficiently initial concentrations of the raw effluent up to 28,400 mg COD/L. PMID- 14524664 TI - Textile dyeing wastewater treatment in a sequencing batch reactor system. AB - This study was undertaken to examine the feasibility of treating biologically textile wastewater for organic carbon removal. The study was conducted over a lab scale SBR equipped with an in-house developed data acquisition and control software. From monitored operation of SBR and dissolved oxygen values, together with a simple compressed air ON/OFF control scheme, on-line Oxygen Uptake Rate (OUR) profiles during aerobic reaction periods were obtained. Due to the high variability of wastewater characteristics, periodical analyses of textile wastewater were conducted and thus characterized for pH, conductivity, total and volatile solids, COD, ammonia, and TKN. After an initial period of activated sludge adaptation to textile wastewater, the SBR was operated at step-feed strategy to reduce the effluent biodegradable matter presents in the wastewater by SBR water change ratio modification and feeding strategy. PMID- 14524665 TI - Treatment of mixed pharmaceutical industry and domestic wastewater by sequencing batch reactor. AB - Raw mixed pharmaceutical industry and domestic wastewater characteristics are such as BOD5 = 90-130 mg L(-1), COD = 200-300 mg L(-1), SS = 900 mg L(-1), pH = 6.4-6.8, T = 20 degrees C, NH3 = 26 mg L(-1), PO4(3-) = 8.5 mg L(-1). In treatment plant, the parameters such as BOD5, NH3, PO4(3-), SS, MLVSS, pH, temperature, sludge volume and microorganisms predominance are studied, under constant sedimentation time (30 min) against variable aeration times. As a second step, the same parameters are studied under the constant aeration time (4 h) against variable sedimentation times. Effluent concentrations from sequencing batch reactor (SBR) are determined BOD5 = 13-18 mg L(-1), COD = 25-37 mg L(-1), SS= 9-21 mg L(-1), pH = 7.3-7.6, TDF = 1-2, T = 23 degrees C, NH3 = 1 mg L(-1), PO4(3-) = 8.1 mg L(-1). At the end of 6 h of aeration time, the efficiency of BOD5 reaches to 83%. The efficiency of BOD5 reaches to 81% under a 4 h aeration time and at 90 min sedimentation time. As a result, the optimum treatment efficiency has been taken under 4 h of aeration time and of 60 min of sedimentation time. PMID- 14524666 TI - Biodegradation of 3-chlorophenol in a sequencing batch reactor. AB - The present paper shows the results obtained through a study on the biodegradation of 3-chlorophenol (3-CP) in a Sequencing Batch Reactor (SBR). To such a purpose a lab-scale SBR was fed a synthetic wastewater containing 3-CP and nutrients (nitrogen and phosphorus) diluted in tap water. The operating strategy, in terms of both the duration of either the cycle or the react phase, was changed throughout the experimental activity in order to find out the optimal one allowing to ensure constant and high removal efficiency despite the increasing 3 chlorophenol concentration in the feed. Biomass collected from a full-scale continuous flow activated sludge facility treating domestic wastewater was used as seed, after being acclimated to 3-CP by means of several batch tests. The results showed that a periodically operated activated sludge system can be successfully used for the biodegradation of chlorophenol compounds, after the needed members of the microbiological consortium are selected and enriched. PMID- 14524667 TI - Design of sequencing batch reactors for biological nitrogen removal from high strength wastewaters. AB - This paper covers an evaluation of more than twenty full-scale industrial wastewater treatment plants employing sequencing batch reactor (SBR) process mainly for carbon removal and a pilot-scale SBR designed for carbon and nitrogen removal from tannery effluent. The study highlights the major features of the SBR technology and proposes a rational dimensioning approach for carbon and nitrogen removal SBRs treating high strength industrial wastewaters based on scientific information on process stoichiometry and modeling, also emphasizing practical constraints in design and operation. PMID- 14524668 TI - Practical aspects of nitrogen and phosphorous removal with floating media SBBR. AB - Practical aspect on the application of floating media for the sequencing batch biofilm reactor (SBBR) has not been studied in detail, especially focused on settling step, nutrient removal and temperature effects. Two types of floating media (sponge and plastic) had been examined for SBBR operation with sewage. Based on the observations with various experimental variables including temperature, media volume, operating methods, both SBBR units with sponge and plastic media generally produced a stable nitrified effluent, but depicted unstable phosphorus removal. The sponge media showed better nitrogen removal, while plastic media showed better phosphorus removal. Overall nutrient removal capability for SBBR was better than a typical SBR without media. In addition, SBBR with sponge media was capable to absorb shock loads, but could not effectively nitrify at a lower temperature even with an increased media volume. In order to produce lower SS effluent as well as minimize P release during the settling step, the floating media SBBR systems would require a dual settling stage. PMID- 14524669 TI - Combined carbonaceous removal and nitrification with biological aerated filters. AB - This paper describes the use of a submerged biological aerated filter (BAFs), on a pilot plant scale, in the secondary nitrification of low strength domestic wastewater. The results, obtained throughout three months of tests, confirmed the ability of BAFs to obtain simultaneous carbonaceous removal and nitrification. In fact, alter a start-up period of 5 weeks, a remarkable steady state condition in the removal of suspended solids (TSS), chemical oxygen demand (COD) and nitrification was reached. For a hydraulic loading rate of 1.6 m3 m(-2) h(-1), the mean efficiency of the process in the removal of TSS and COD was 85% and 83% respectively. Mean concentrations in the influent flow of around 114 mg(TSS)/L and 140 mg(COD)/L were reduced to average values of below 17 mg(TSS)/L and 25 mg(COD)/L. Secondary nitrification was achieved with a mean efficiency of 85% for a mean volumetric loads of 0.45 kg(N-Namm) m(-3) d(-1) (25 degrees C). Through an intensive activity of nitrification, in the steady state condition, mean influent concentrations of around 12 mg(N-Namm)/L were reduced to average values of below 2 mg(N-Namm)/L. The removal loading rate increased linearly with the applied loading rate for volumetric loads of up to 0.7 kg(N-Namm) m(-3) d(-1). The nitrification was not sensitive to the applied COD loads because of the low level of carbon substrate. PMID- 14524670 TI - Nitrification-denitrification of opto-electronic industrial wastewater by anoxic/aerobic process. AB - This research focused on the biological treatment of high-strength organic nitrogen wastewater, and presented the results from the nitrification and denitrification of an actual industrial wastewater using anoxic/aerobic process. The opto-electronic industrial wastewater often contains a significant quantity of organic nitrogen compounds and has a ratio over 95% in organic nitrogen (Org N) to total nitrogen (T-N). In this study, a 2-stage anoxic/aerobic process was established and evaluated the efficiency of wastewater treatment. Raw wastewater from an actual TFT-LCD manufacturing plant was obtained as the sample for looking into the feasibility of opto-electronic industrial wastewater treatment. After toxicity identification evaluation (TIE) test of raw wastewater. the inhibition was related to organic nitrogen (TMAH, MEA) and unionized ammonia (free ammonia, NH3) with high pH. Therefore, pH control is important for biological treatment of high-strength organic nitrogen industrial wastewater. Besides. hydraulic retention time (HRT) and mixed liquor recycled rate (MLR) were controlled independently to distinguish between the effects of these two factors. Under suitable HRT ( > 1.7 d) and mixed liquor recycled rate (< 4Q), effluent of NH4-N. NO3-N + NO2-N, and COD can fall below 20 mg/L, 30 mg/L, and 80 mg/L. The anoxic/aerobic process removed 92-98% of the carbon source, and approximately 80% of TKN, 70% of T-N. PMID- 14524671 TI - Denitrification in tertiary filtration: application of an up-flow filter. AB - The present paper shows the results obtained through an experimental work performed at the wastewater treatment plant of Rome, aimed at studying the performances of a tertiary filter regarding combined removal of suspended solids, COD, and nitrates. The up-flow sand filter was fed by the effluent coming from the secondary settling tank of the plant. The filter bed height was of 80 cm of silica sand. After a start up period, a study of particulate and soluble COD removal process was made, to establish the need of methanol in the denitrification process. Total COD removal efficiency was 60% on average, 55% due by soluble COD removal and 5% by particulate one. In the last phase of the experimental activity methanol was fed as carbon source, sodium sulfite was supplied to produce anoxic environment within the filter and the denitrification efficiency was studied. Nitrates removal rates after an acclimation period of 10 days increased up to 60%, with an effluent NO3-N of 8 mg/L. Denitrification rate was 2.4 kg/m3 d for water temperatures of 25 degrees C. Regarding methanol demand and biologic kinetics, the biomass yield coefficient was 0.3 kg(COD-X)/kg(me). Consequently 2.7 kg of methanol was required per kilogram of denitrified nitrogen. PMID- 14524672 TI - Novel phased isolation ditch system for enhanced nutrient removal and its optimal operating strategy. AB - Phased isolation ditch system with intrachannel clarifier is a simplified novel oxidation ditch system enhancing simultaneous removal of biological nitrogen and phosphorus in municipal wastewater. The system employs two ditches with intra clarifier, and eliminates external final clarifier, additional preanaerobic reactor, and recycle of sludge and nitrified effluent. Separation of anoxic, anaerobic, and aerobic phases can be accomplished by alternating flow and intermittent aeration. Its pilot-scale system operated at HRTs of 10-21 h, SRTs of 15-41 days, and a cycle times of 2-8 h showed removals of BOD, TN, and TP in the range of mixed liquor temperature above 10 degrees C as high as 88-97, 70-84, and 65-90%, respectively. As the SRTs became longer, the effluent TN decreased dramatically, whereas the effluent TP increased. Higher nitrogen removal was accomplished at shorter cycle times, while better phosphorus removal was achieved in longer cycle times. Optimal system operating strategies maximizing the performance and satisfying both the best nitrogen and phosphorus removals included HRTs ranged 10-14 h, SRTs ranged 25-30 days, and a cycle time of 4 h at the mixed liquor temperature above 10 degrees C. Thus, complete phase separation in a cycle maximizing phosphorus release and uptake as well as nitrification and denitrification was accomplished by scheduling of alternating flow and intermittent aeration in the simplified process scheme. Especially, temporal phase separation for phosphorus release without additional anaerobic reactor was successfully accomplished during anaerobic period without any nitrate interference and carbon-limiting. PMID- 14524673 TI - The role of DPB at anoxic stage in a novel package type BNR process with batch settler. AB - In this research, package type BNR process was developed to remove nutrients as well as suspended solids from domestic sewage. The effect of HRT reduction of settler on the removal of pollutants as well as the role of DPBs at anoxic reactor were investigated. The proposed package system was composed of sludge denitrification tank, anaerobic, anoxic, oxic, and batch settler. This system could remove nitrogen and phosphorus effectively at low COD/N ratio and also remove SS more effectively than other ordinary BNR system having the conventional settlers. The removal efficiency of total nitrogen (T-N) in optimal condition was about 75.1% under the TCOD/TKN ratio as low as 5.7. The average concentration of the effluent TCOD, ortho-P, NH4+-N, NO3- -N and SS in the package type BNR system of 2 h settler retention time were 15.6. 1.38, 1.4, 10.3 and 3.9 mg/L, respectively. In anoxic state, denitrifying dephosphatation, that is uptaking phosphate by using nitrate-nitrogen as an electron acceptor was observed. The removed NO3- -N concentration by denitrifying dephosphatation was 1.62 mg NO3- N/mg PO4(3-)-P. PMID- 14524674 TI - Combined physical-chemical and biological treatment of poorly biodegradable industrial effluents. AB - Effluents from small and medium sized chemical plants may contain significant amounts of poorly biodegradable aromatic compounds, which could negatively affect water quality and public health. This is a key environmental issue, particularly in areas where effluents are discharged into drinking water sources. Unfortunately, conventional biological treatment may not be able to meet discharge standards, and combined systems should be implemented. In this context, this paper presents experimental results on the application of a combined sequential ozonation-activated carbon-biological system to treat effluents containing chlorinated aromatic contaminants from chlorine based pulp bleaching. The experimental system consisted of an ozone bubble column reactor (0.3 dm3), an activated carbon fixed bed reactor (0.2 dm3), and an aerobic bioreactor (20 dm3). Ozone was produced from pure O2 using a generator rated at 2 mmol O3 h(-1). The bleaching effluent was pretreated and fed into the aerated sequencing batch bioreactor containing preconditioned biological sludge (3-4 g VSS dm(-3)), and cultured for 24 h. Samples of raw and treated effluents were assayed for biochemical oxygen demand (BOD5), chemical oxygen demand (COD), total organic carbon (TOC), total phenols, and adsorbable organic halogens (AOX), using standard techniques. The presence of potential genotoxic activity in untreated and treated samples was assessed using the Ames tests. Results show that biological treatment of raw samples could not remove mutagenic activity on its own. On the other hand, ozonation followed by activated carbon treatment and biological treatment successfully removed genotoxicity in all cases. Reductions in BOD, COD, TOC, AOX, and phenols by biological treatment increased when samples were pretreated with ozone/activated carbon. PMID- 14524675 TI - Comparative evaluation of the results for the synthetic and actual reactive dye bath effluent treatment by nanofiltration membranes. AB - In this study, the results of nanofiltration experiments carried out on reactive dye house wastewaters are presented. The experiments were performed using lab scale membrane system. Permeate quality was appropriate to reuse permeate in dyeing process again. Pretreatment and neutralization were the parameters which are very important for the recovery of high amounts of salt and water at the permeate stream. In addition, use of HCl instead of H2SO4 for neutralization increased the NaCl recovery. PMID- 14524676 TI - Treatment of Cr(VI) and phenol by illuminated TiO2. AB - Industrial wastewater from organic chemical manufacturing and metal plating industries may contain significant amounts of refractory organic matter and heavy metals. Heavy metals such as Cr(VI) and refractory organic matter like phenol were simultaneously removed by reduction and mineralization, respectively, during photocatalytic process. Therefore, this study was carried out to evaluate the effects of phenol on the removal of Cr(VI) in several environmental conditions. Cr(VI) and phenol were more effectively eliminated in UV/TiO2 condition than either by UV or TiO2-alone. The removal efficiencies of Cr(VI) increased as the pH of the TiO2 slurry decreased, the concentration of phenol increased up to 10 mg L(-1), and nitrogen gas was used. Otherwise, the removal efficiencies of phenol were higher at O2-purged system and neutral pH. Without regard to purging gases, complete Cr(VI) removal was observed at pH 4 after 80 min while 90% phenol destruction and 60% mineralization were found after 120 min. It wasfound that application of photocatalytic reaction by using illuminated TiO2 to treat wastewater containing both Cr(VI) and phenol was possible. PMID- 14524677 TI - Advanced oxidation of biologically pretreated Baker's Yeast Industry effluents for high recalcitrant COD and color removal. AB - The aim of this study was to investigate the effectiveness of chemical oxidation by applying ozonation, ozonation with hydrogen peroxide and Fenton's processes for decolorization and residual COD removal of biologically pretreated baker's yeast industry (BYI) effluents. Baker's yeast industry effluents characterizing with high COD, TKN, dark color, and non-biodegradable organic pollutants. The batch tests were performed to determine the optimum operating conditions including pH, O3, H2O2, and FeSO4 dosages, molar ratio of Fe2+/H2O2 and reaction time. It was noticed that H2O2 significantly reduced the reaction times for the same ozone dosages: however, COD and color removals were not remarkable. In the Fenton's oxidation studies, the removal efficiencies of COD and color for 30 min reaction time for three different types of BYI effluents were found about 86 and 92%, respectively. Experimental results of the presented study have clearly indicated that the Fenton's oxidation technology is capable to fate almost all parts of the organics which consist of both soluble initial and microbial inert fractions of COD for baker's yeast effluents. Effluents from the Fenton's oxidation process can satisfy effluent standards for COD and color in general. PMID- 14524678 TI - Removal of color and COD from a mixture of four reactive azo dyes using fenton oxidation process. AB - This study was designed to decolorize and to reduce COD content in a mixture of four reactive dyes, i.e., Remazol Black 5 (RB5), Remazol Red RB (RR), Remazol Yellow 84 (RY), Remazol Brilliant Blue (RB) using Fenton Oxidation Process (FOP). Optimum pH, temperature, and the doses of FeSO4 and H2O2 were determined. Experiments were conducted on the samples containing a total concentration of 100 mgL(-1) (RB+RY), 200 mg L(-1) (RB5+RR), 300 mg L(-1) (RB5+RR+RB+ RY), and 400 mg L(-1) (RR+RB+RY) dyes considering their actual application doses in dyehouses. Optimum pH was observed as 2.5 at 30 degrees C using 400 mg L(-1) FeSO4 and 800 mg L(-1) of H2O2 resulting in more than 96% COD and 99% Pt-Co unit of color removal for the mixture of RB5 and RR. The optimum conditions determined were 4.0 pH, 50 degrees C, and 500 mg L(-1) FeSO4 applying 1000 mg L(-1) H2O2 for the mixture of (RB5+RR+RB+RY). A 100 mg L(-1) solution of a mixture of RB and RY at equal amounts was oxidized using 200 mg L(-1) FeSO4 and 300 mg L(-1) H2O2 at 3.0 pH and 50 degrees C. PMID- 14524679 TI - Clay mineral batch process for color removal of textile wastewaters. AB - Textile industry effluents exhibit large amounts of dye chemicals, which create severe water pollution. It is therefore important to reduce the dye concentration in the wastewater before discharging into the environment. In this study, the ability of sepiolite to uptake different reactive dyes is investigated. The dyes used in the experiments are Everzol Black B, Everzol Yellow 3RS H/C, and Everzol Red 3BS. A number of parameters on adsorption has been investigated in order to establish the adsorption pattern of sepiolite. The effect of conditioning time, solid/liquid ratio, pH, temperature, and dye concentrations on adsorption are studied. Adsorption isotherms for different reactive dyes have been obtained using both natural and modified sepiolites. The maximum capacity of natural sepiolite for the reactive dyes occurs at around 0.1-1 mg/g. Therefore, the natural sepiolite appears to have a little capacity for reactive dyes. The zeta potential profiles of natural and modified sepiolites against pH are also examined. The modified sepiolite exhibited ten times higher performance than the natural sepiolite. PMID- 14524680 TI - Photochemical degradation and mineralization of phenol: a comparative study. AB - In this study, photochemical advanced oxidation processes (AOPs) utilizing the combinations of UV, UV/H2O2, and UV/H2O2/Fe2+ (Photo-Fenton process) were investigated in lab-scale experiments for the degradation and mineralization of phenol. The major parameters investigated were the initial phenol concentration, pH, hydrogen peroxide, and iron doses, and the effect of the presence of radical scavengers (PO4(-3), SO4(-2), and Cl- ions). It was observed that the phenol degradation efficiency decreased with increasing phenol concentration and pH in UV process. Maximum phenol oxidation efficiency for an initial concentration of 100 mg L(-1) and at pH 3 was around 30% in direct photolysis. The efficiency increased to 97% with UV/H2O2 process, however, there was still negligible mineralization (9%) and the required irradiation time was still long (300 min). The results showed that the Photo-Fenton process was the most effective treatment process under acidic conditions. Complete disappearance of 100 mg L(-1) phenol was achieved in 2.5 min and almost complete mineralization (97%) was also possible after 300 min of irradiation. The efficiency was negatively affected from H2O2 in UV/H2O2 process and Fe2+ in Photo-Fenton process over a certain concentration. The highest negative effect was observed with solution containing PO4(-3) ions. Required reaction time for complete disappearance of 100 mg L(-1) phenol increased from 2.5 min for an ion-free solution to 60 min for that containing PO4(-3). The photodegradation of phenol was found to follow the first order law. PMID- 14524681 TI - Design and performance of magnetic composite particles for the separation of heavy metals from water. AB - Composite particles have been made by combining nanosized polymer particles with magnetite microparticles using a ball-mill process. The magnetite particle cores were included to facilitate the composite particles separation by use of a magnetic field. Polymer particles carried carboxylic groups as ion-exchange vehicles. The composite particles were used in the extraction/stripping of Zn, Cu, Ni, and Cr from aqueous solutions in a stirred batch reactor of laboratory scale. The extraction behavior of these metals was studied as a function of initial metal concentrations of up to 25 mM, ionic strength at pH (5-7), and temperature (1-80 degrees C). Experimental results showed that the process was very fast and yielded complete metal removal when the initial concentrations of metals were very low. The selectivity series was Cu > Cr > Zn > Ni. The effect of ionic strength was immaterial at pH 7. However, extraction decreased with increasing ionic strength at pH 5. Extraction increased with increasing temperature. A series of extraction/ stripping experiments indicated that the particles can be reused without significant loss in their extraction capacity. PMID- 14524682 TI - Performance-based characterization of a contact stabilization process for slaughterhouse wastewater. AB - This paper evaluates the effluent treatment plant of a slaughterhouse in Hawalli City, Kuwait processing 1100 heads of livestock a day. Results indicated that the proposed process effectively reduces pollution potential of slaughterhouse wastewater. Influent Chemical Oxygen Demand (COD) ranged from 3335 to 7580 mg L( 1), of which approximately 30% were in the form of suspended solids (SS). Removal efficiency was 77% for soluble COD and 82% for insoluble COD, at a volumetric load of 1.8 kg COD m(-3) d(-1). Values obtained for the biokinetic coefficients, floc uptake (FU), substrate removal efficiency (SRE), specific reaction rate (RR), maximum reaction rate (Rm) and Yield (Y) of the contact tank were higher than the range of values reported for other continuously fed activated sludge systems. In contrast to the contact tank, the aeration basin biokinetic coefficients were within the range of values reported. Contact process testing demonstrated that controlling solid recirculation to maintain a contact loading of about 120-150 mg COD g(-1) VSS in the contact tank generally resulted in high SRE. RR, Rm, and also in good settleability as indicated by SVIs being consistently below 150 mL g(-1). In the other hand, higher contact loadings of more than 150 mg COD g(-1) VSS. resulted in a significant deterioration in SRE, RR, Rm, and SVIs. PMID- 14524683 TI - Aromatic amine degradation in a UASB/CSTR sequential system treating Congo Red dye. AB - In this study an anaerobic (upflow anaerobic sludge blanket reactor)/aerobic (completely stirred tank reactor) sequential system was used to treat a synthetic wastewater with minerals and co-substrate together with 100-4000 mg L(-1) of Congo Red dye (Direct red 28) (CR), which is a banned azo dye in Turkey. The effect of hydraulic retention time (HRT) on the decolorization and the COD removal efficiency was investigated at constant 100 mg L(-1) Congo Red concentration. 77% of COD and 95% of color was removed at a HRT of 0.486 days and a maximum organic loading rate of 6.656 kg COD m(-3) day(-1) in the anaerobic/aerobic stage. In the continuous operations, 88% of COD, 99% of color and 91% of total aromatic amine (TAA) were removed at a HRT of 3.60 days and at a CR concentration of 4000 mg L(-1). This corresponds to an organic loading rate of 1.81 kg COD m(-3) day(-1), and a CR dye loading rate of 46.37 g dye m(-3) h(-1), respectively, in the whole system. The TAA produced under anaerobic conditions was ultimately removed in the aerobic stage, resulting in very low aromatic amine recoveries (5-18%) in the last one. Therefore the aerobic effluents exhibited higher IC50 and specific methanogenic activities (SMA) compared to anaerobic and dye containing samples, indicating the reduced toxicity. PMID- 14524684 TI - Upgrading of a small overloaded activated sludge plant using a MBBR system. AB - The aim of this research was the application of a biofilm system for the upgrading of a full-scale overloaded activated sludge MWWTP using the MBBR (Moving Bed Biofilm Reactor) technology. The choice of this fixed biomass system appeared appropriate because it offers several advantages including good potential in nitrification process, easiness of management and above all, the possibility to use the existing tank with very few modifications. MBBR system counts only few full-scale plants in Italy at the moment, thus a pilot-scale experimentation was preliminarily carried out. The acquired parameters were used for the fullscale MWWTP upgrading. The upgrading of the activated sludge reactor in the MBBR system has given (1) a relevant increase in the flowrate treated up to 60%; (2) a good efficiency in organic carbon removal and nitrification, equal to 88% and 90% respectively, with HRTs of 5.5-7 h; (3) the overcoming of the hydraulic overload of the secondary settler, applying a lamellar settler. It was observed a good correlation between the results obtained at pilot-scale and those observed in the full-scale plant. PMID- 14524685 TI - Operational strategies for a small wastewater treatment plant using OUR, microscopic analysis, and toxicity test. AB - This study was focused on determining the optimal operational conditions of a small domestic wastewater treatment plant (SWP) with a flow rate of 1000 m3 d(-1) using oxygen uptake rate (OUR) data, microscopic analyses, and effluent toxicity testing with Daphnia magna in parallel to wastewater characterization. Chemical analyses of nine samples, taken from different points of the treatment plant over a nine months period, presented strong domestic wastewater characteristics. The plant performance for meeting discharge standards was not sufficient enough for TP parameter. Aeration capacity and sludge recycle ratio were adjusted to improve the performance of the system and to prevent any operational problems, such as bulking. Although the inflow was 100% toxic, effluent toxicity following treatment was decreased to 5%. Filamentous bacteria were in "excessive" amount. The dominant filamentous bacteria species were determined as Nostocoioda limicola II. Reevaluation of the discharge limits with respect to nutrient removal, geographic location, and receiving water quality subject to treatment effluent discharges. PMID- 14524687 TI - A high-rate secondary treatment based on a moving bed bioreactor and multimedia filters for small wastewater treatment plants. AB - For small wastewater treatment plants (WWPTs), high-rate secondary treatment systems with good treatment efficiency and easy, stable, and robust operation are called for. In this paper an experimental study on a high rate secondary treatment based on moving bed biofilm reactor (MBBR) and multimedia filters is presented. A high rate MBBR converts easily biodegradable SCOD in short HRT (0.5 h) directly after screening, then a Kaldnes-Filtralite-Sand (KFS) filter removes the particulate COD and detached biofilms at filtration rates of 10-20 m/h. The whole system gave effluent SS and COD less than 30 mg/L and 100 mg/L when total detention time is less than 1 h and small dosage of chemicals (iron and/or cationic polymer) is used. A new scenario of high rate secondary system with a primary Kaldnes coarse media filter in front of high rate MBBR and the KFS filter is proposed and discussed. This scenario with total HRT less than 2h is more suitable for high influent SS concentrations and may also be extended for nitrogen and phosphorous removal. Compared to conventional secondary treatment, the high rate secondary treatment will be using only 1/5-1/10 of the space, resulting in considerable savings for construction, energy and operation. PMID- 14524686 TI - Recycling and reuse of wastewater from a new-developed community using sand filtration, ultrafiltration, and ozonation. AB - In this study, the pilot apparatus combined with sand filtration, ozonation and ultrafiltration was established. Wastewater from the secondary treatment effluent in the new-developed community was taken as the sample for looking into the feasibility of domestic wastewater reuse and recycling. The test results by sand filtration, sand filtration/ultrafiltration, sand filtration/ozonation, and sand filtration/ultrafiltration/ozonation were compared for looking for appropriate treatment processes applied in the domestic wastewater reuse and recycling. Finally, cost analysis was carried out and sand filtration/ozonation process was suggested to be one of the best processes. The total cost is about 0.1-0.32 USD dollars per cubic meter of produced water by considering the capital and operation cost for five years in the small domestic wastewater treatment plant (50-750 CMD). PMID- 14524688 TI - Development of floating plastic media filtration system for water treatment and wastewater reuse. AB - Floating plastic media coupled with sand filtration system was applied for treating surface water and secondary effluent from municipal sewage treatment plant. The system employed floating plastic media for the removal of suspended solids in surface water and precipitated phosphorus from secondary effluent of sewage treatment plant. Sand filtration was then used to remove further the suspended solids. For the purposes of wastewater reuse, a zeolite layer was used instead of sand filter to absorb ammonium nitrogen. Application of system for surface water treatment suggested appropriate filtration rate of 5 m3/m2 h. Polyaluminum chloride was found to be the best coagulant with an appropriate plastic and sand bed depth of 40 cm. The system could produce average effluent turbidity and suspended solids of 0.71 NTU and 0.94 mg/L respectively. Average turbidity and suspended solids removal efficiencies were 96.26% and 95.48% with low headloss development of 40.4 cm at the end of 6 h operation period. When applying 1.50 m. floating plastic media bed for the treatment of synthetic raw water, short and long-term turbidity removal efficiencies were 96.79-97.72% and 81.81-94.61% for raw water containing turbidity of 20 and 40 NTU. It could produce the effluent with turbidity less than 5NTU while having less than 1.0 m. headloss. The system was also applied for the secondary effluent treatment. An optimum filtration rate of 5 m3/m2 h was obtained when using plastic and sand bed depth of 60 and 20cm under direct filtration mode. Average turbidity removal was 60.3% and 59.6% after 6 and 48 h of operation. It was also found that 1 m3/m2 h filtration rate and 50:30 cm of plastic:zeolite bed could be used to achieve both ammonium nitrogen and phosphate removal. Suspended solids, turbidity, BOD5, NH4+, and PO4(3-) removal efficiencies were 91.9, 94.6, 95.4, 97.3, and 99.5% respectively after 24 h. As a result, the effluent from the system had average NH4+ and PO4(3-) of 0.5 mg N/L and 0.02 mg P/L. PMID- 14524690 TI - Two stage thermophilic anaerobic-aerobic mineralization-stabilization of excess activated sludge. AB - At the National Institute of Chemistry, Ljubljana, Slovenia, new procedure for excess activated sludge was developed and patented. The main principle of the sludge mineralization is two-step process--the first is anaerobic, and the second step is aerobic. In the first step maximal biogas production is acquired in the second step at smaller VSS the optimal VSS and COD is achieved. Anaerobic, aerobic, and combined sludge mineralization was studied. The next combinations of different successive anaerobic and aerobic mineralization have been studied: 3 + 3 (3 days of anaerobic + 3 days of aerobic digestion), 3 + 6, 5 + 5, 3 + 12, and 10 + 10 days, respectively. The best combination considering biogas production, VSS and COD reduction have been gained for the combination of 3 + 6 days, and 3 + 12 days. At 3 + 6 days digestion about 49% of VSS, and about 51% of COD reduction was gained, at 3 + 12 days about 62% of VSS, and about 57% of COD reduction was achieved. For the optimal system operation, economics, biogas production, and necessary VSS and COD reduction have to be considered. PMID- 14524689 TI - Anaerobic digestion of municipal sludges with high silt content using granular seed. AB - The performance of anaerobic digestion of municipal sludge having high inorganic solid/silt in Tuzla Wastewater Treatment Plant (TWWTP) in Istanbul was evaluated using granular seed. High silt/solid content is the main problem related to wastewater collection system in TWWTP. Due to high TSS concentration of the influent, inorganic solid ratio reaches to 60-70% in the primary sludge that makes sludge digestion very difficult. The aim of this study was to evaluate the detrimental effects of inorganic silt/clay on anaerobic sludge solubilization of the primary sludge (PS), of the biological sludge (BS), and of the mixed sludge (50%PS + 50%BS). Anaerobic solubilization was carried out in continuously stirred anaerobic reactors at mesophilic temperature (35 degrees C). Results indicated significant total biogas productions in all reactors during digestion period. The biogas production of the primary sludge was higher than the biogas productions of the mixed sludge and the biological sludge. The methane contents of the primary, biological. and mixed sludges were found as around 72, 78, and 75%, respectively. When all types of sludges were used as substrates, the reactors removed VSS with a corresponding production of Soluble COD (SCOD). The greatest degree of hydrolysis/acidogenesis was observed in all types of sludges in about 10 days of operation and became almost stable which indicated that the reactors were still in the hydrolysis/acidogenesis phase. VFAs measurements were well matched with SCOD results. In all reactors, observation of the conversion into methanogenic phase in about 1.5 month was not possible which might be attributed to high inorganic silt/clay and salinity content of TWWTP. High inorganic silt/clay and salinity might have decreased the biodegradability of all sludges and methanogenesis could not operate in early periods of digestion. Higher SCOD decreases might have been possible if longer treatment time and more optimal conditions for anaerobic solubilization were applied. The pH did not fall below the unfavorable range in all reactors during the study. PMID- 14524691 TI - Effect of sludge water distribution on the liquid-solid separation of a biological sludge. AB - Water distribution in sludge is worthy of investigation due to its importance in the liquid-solid separation of sludge. Biological sludge contains different physical states of water, including free water, interstitial water, vicinal water, and water of hydration, associated with sludge solids. Conventional dewatering processes can remove only the free water and a part of the interstitial water. Release of interstitial water held inside the cell structure requires the disruption of sludge cells. This study investigates the effect of water distribution on the liquid-solid separation of a biological sludge. Sludge cells were disrupted by alkali treatment, NaCl treatment, heat treatment, and sonication. Water distribution in sludge was determined by differential scanning calorimetry (DSC) and centrifugation. The results of this study indicated that the sludge cell disruption changed the water distribution in sludge and improved its compactibility (as a measure for liquid solid separation). It released a considerable amount of interstitial water trapped inside the cells. Cell disruption created extra surfaces for water binding and increased the unfreezable water content (mostly vicinal water, water of hydration, and a fraction of interstitial water). Compactibility (liquid-solid separation property) of the sludge in terms of cake solids content was improved to about 2-7.5 folds depending on the disruption method used. PMID- 14524692 TI - Sludge dewatering and disposal practices for small activated sludge wastewater treatment plants. AB - Sludge dewatering is a decisive step in the reduction of waste sludge volume, thus considerably affecting total sludge treatment and disposal costs. The construction of sludge dewatering facilities in small WwTPs though, is generally not cost effective. In this paper some experimental evidence is presented, that waste sludge dewatering in small WwTPs of the activated sludge extended aeration type, can be effectively achieved by a centrifuge type of equipment withdrawing sludge directly from the aeration tank; an economic evaluation of the possibility to employ a transportable type of similar equipment mounted on a truck, to serve a number of small WwTPs located in remote or isolated areas is also presented and discussed. PMID- 14524693 TI - Plant availability of heavy metals and phosphorus in the agricultural reuse of biological sludges. AB - Concerns about the agricultural use of sludges are usually associated with heavy metals. In this study, plant-availabilities of heavy metals and phosphorus (P) in different P amendments were investigated. The dewatered sludge from Pasakoy Biological Nutrient (N, P) Removal (BNR) Plant in Istanbul, a chemical fertilizer (Triple Superphosphate), and a P-deficient, alkaline soil were used in pot experiments. Unstabilized sludge (S), lime-stabilized sludge (LS), and triple superphosphate (TSP) were applied to pots in a manner to supply 0, 100, 200, 300, 400 ppm P. Lollium Perenne was grown on these soils to compare P and heavy metal (Cu. Ni, Zn) availabilities in terms of plant uptake. The results of plant analyses showed that P-recycling through the agricultural use of biological nutrient removal (BNR) sludges was possible. Plant availability of heavy metals in sludge amended soil samples was comparable with those in control samples (soil only) and chemical fertilizer amended samples. Lime stabilization reduced the plant-availability of P and heavy metals. The choice of lime stabilization must depend on the specific soil to be amended. PMID- 14524694 TI - Sludge dewatering using macrophytes in a small wastewater treatment system: a case study of a pilot scale plant in northern Italy. AB - A study was conducted on a system consisting of two beds of 12 m2 each, planted with reeds and filled with sludge from an activated sludge plant dimensioned for 3000 p.e. During 2001, 10 kg of TS/m2 were loaded in each bed using sludge with a dry matter content of 3% during winter and of 0.5% in summer. The aim of the study was to evaluate parameters such as: COD, TSS, P-PO4(3-), N-NO2-, N-NO3-, N NH4+ in percolation water, the dewatering capability of the two beds and to estimate N mass balance. The observations on dewatering process showed that dried sludge reached a content of 23% TS in winter and of 30% TS in summer. During summertime the plant did not release any percolation flow; in the rest of the year the concentrations of COD, TSS, P-PO4(3-), N-NO2-, N-NH4- remained within bounds of the European directive (91/271/CEE). The N mass balance proved that 90% of N was removed by the denitrification process. PMID- 14524695 TI - Merits of polyoxotungstates as environmental remediation catalysts: a novel wet oxidation technology for refractory industrial pollutants. AB - The catalytic effect of silicadodecatungstate (SiW12O40(4-)) on the oxidation of nitrobenzene in subcritical water (T = 100-300 degrees C; P = 0.1-8.6MPa, pH = 2.6) was examined. In the absence of SiW12O40(4-) catalyst nitrobenzene oxidation slowed down significantly after a brief period of rapid degradation and an average of 40% nitrobenzene removal. The catalytic effect of SiW12O40(4-) becomes particularly pronounced after a temperature dependent induction period. Catalyzed wet air oxidation continued until the degradation of the parent compound was complete provided that experimental conditions were harsh enough (T > 200 degrees C). Product distribution analysis suggested that the oxidation paths for catalyzed and uncatalyzed wet air oxidation of nitrobenzene are similar, but the analysis was not definitive. PMID- 14524696 TI - Capacity of the bioremediation technology for clean-up of soil and groundwater contaminated with petroleum hydrocarbons. AB - A column reactor was designed and used to simulate conditions affecting the bioremediations of petroleum hydrocarbons. The work illustratively describes the aerobic (model) clean-up of soil samples enabling to predict the efficiency of a technology installed in parallel on contaminated former airport. The data showing the performance of thus precharacterized technology are presented. PMID- 14524697 TI - The effects of diquat dibromide on biological wastewater treatment plants. AB - The objective of the study was to investigate the fate and effects of diquat dibromide which is the active ingredient in formulations used to control the growth of roots into sewers when applied as Razorooter and mixed with raw sewage, settled sewage, and activated sludge, and when introduced into activated sludge wastewater treatment systems. Both fully aerobic and biological nutrient removal (BNR) activated sludge systems were used for experimental purpose, and both continuous flow and batch reactors were used. The sorption of diquat by both raw sewage particles and activated sludge suspended solids was determined. Diquat dibromide concentrations ranged from 0.93 to 12.6 mg/L in the influent flow. Both the fully aerobic and two full biological nutrient removal systems were fed municipal sewage spiked with diquat dibromide, and operated at a mixed liquor temperature of 10 degrees C and an MCRT of 10 days. One of the BNR systems was a control system. The results showed that only about 20% of the diquat in raw sewage flow was removed by adsorptions to the sewage solids, but 80% or more of the diquat was removed in activated sludge systems. When the influent diquat dibromide concentration was approximately 1mg/L, over 99% of the diquat dibromide was removed by the activated sludge process. Some of the removal was believed to be by biodegradation. The diquat dibromide used in this study had no observable detrimental effects on any of the biological processes of the continuous flow fully aerobic and BNR activated systems. PMID- 14524698 TI - Perspectives for the industrial enzymatic production of glycosides. AB - Glycosides are of commercial interest for industry in general and specifically for the pharmaceutical and food industry. Currently chemical preparation of glycosides will not meet EC food regulations, and therefore chemical preparation of glycosides is not applicable in the food industry. Thus, enzyme-catalyzed reactions are a good alternative. However, until now the low yields obtained by enzymatic methods prevent the production of glycosides on a commercial scale. Therefore, high yields should be established by a combination of optimum reaction conditions and continuous removal of the product. Unfortunately, a bioreactor for the commercial scale production of glycosides is not available. The aim of this article is to discuss the literature with respect to enzymatic production of glycosides and the design of an industrially viable bioreactor system. PMID- 14524699 TI - Expression of a Phanerochaete chrysosporium manganese peroxidase gene in the yeast Pichia pastoris. AB - A gene encoding manganese peroxidase (mnp1) from Phanerochaete chrysosporium was cloned downstream of a constitutive glyceraldehyde-3-phosphate dehydrogenase promoter in the methylotrophic yeast Pichia pastoris. Three different expression vectors were constructed: pZBMNP contains the native P. chrysosporium fungal secretion signal, palphaAMNP contains an alpha-factor secretion signal derived from Saccharomyces cerevisiae, and pZBIMNP has no secretion signal and was used for intracellular expression. Both the native fungal secretion signal sequence and alpha-factor secretion signal sequence directed the secretion of active recombinant manganese peroxidase (rMnP) from P. pastoris transformants. The majority of the rMnP produced by P. pastoris exhibited a molecular mass (55-100 kDa) considerably larger than that of the wild-type manganese peroxidase (wtMnP, 46 kDa). Deletion of the native fungal secretion signal yielded a molecular mass of 39 kDa for intracellular rMnP in P. pastoris. Treatment of the secreted rMnP with endoglycosidase H (Endo H) resulted in a considerable decrease in the mass of rMnP, indicating N-linked hyperglycosylation. Partially purified rMnP showed kinetic characteristics similar to those of wtMnP. Both enzymes also had similar pH stability profiles. Addition of exogenous Mn(II), Ca(II), and Fe(III) conferred additional thermal stability to both enzymes. However, rMnP was slightly less thermostable than wtMnP, which demonstrated an extended half-life at 55 degrees C. PMID- 14524700 TI - Strategic selection of hyperthermophilic esterases for resolution of 2 arylpropionic esters. AB - Homologues to Carboxylesterase NP and Candida rugosa lipase, used for the chiral separation of racemic mixtures of 2-arylpropionic methyl esters, were identified by BLAST searches of available genome sequences for hyperthermophilic microorganisms. Two potential candidates were identified: a putative lysophospholipase from Pyrococcus furiosus (Pfu-LPL) and a carboxylesterase from Sulfolobus solfataricus P1 (Sso-EST1). Although both enzymes showed hydrolytic preference toward the (S) methyl ester, only Sso-EST1 yielded highly optically pure (S) naproxen (%ee(p) >/= 90) and was thus further investigated. Changes in pH or reaction time showed little improvement in %ee(p) or E values with Sso EST1. However, the addition of 25% methanol resulted in a 25% increase in E. The effect of various cosolvents on the enantiomeric ratio showed no correlation with the log P or dielectric constant values of the solvent. However, an inverse relationship between E and the denaturation capacity (DC) of the water miscible cosolvents was observed. This was attributed to an increase in enzyme flexibility with increasing solvent DC values leading to a concomitant reduction in the resolving power of Sso-EST1. The results here show that although bioinformatics tools can be used to select candidate biocatalysts for chiral resolution of 2 arylpropionic esters, biochemical characterization is needed to definitively determine functional characteristics. PMID- 14524701 TI - Production of podophyllotoxin using cross-species coculture of Linum flavum hairy roots and Podophyllum hexandrum cell suspensions. AB - Novel cross-species coculture systems using Linum flavum hairy roots and Podophyllum hexandrum cell suspensions were applied for in vitro production of podophyllotoxin. The hairy roots and suspensions were cocultured in Linsmaier and Skoog medium in dual shake flasks and dual bioreactors. In separate experiments, coniferin feeding was shown to be an effective strategy for increasing the accumulation of podophyllotoxin in P. hexandrum suspensions. Because roots of L. flavum are a natural source of coniferin, hairy roots of this species were used in coculture with P. hexandrum to provide an in situ supply of coniferin. Compared with P. hexandrum suspensions cultured alone in shake flasks or bioreactors, podophyllotoxin concentrations in cocultured P. hexandrum cells were increased by 240% and 72% in dual shake flask and dual bioreactor systems, respectively. The availability and stability of coniferin in the medium are the most likely factors limiting podophyllotoxin synthesis in coculture. Intensification of the coculture process is required to further improve total podophyllotoxin accumulation on a volumetric basis. PMID- 14524702 TI - Tuning the transcription rate of recombinant protein in strong Escherichia coli expression systems through repressor titration. AB - The main goal of this work was to develop a strategy that enables tuning of recombinant gene expression relative to the metabolic capacity of the host cell synthesis machinery. In the past, strong expression systems have been developed in order to maximize recombinant gene expression. However, these systems exert an extremely high metabolic burden onto the host cell, which may even lead to cell death. Hence, the period of recombinant gene expression is significantly reduced, and therefore, maximal yield cannot be attained. To extend the production phase and to achieve optimal yields, adjustment of recombinant gene expression by modulation of the transcription rate is required. To control transcription, we designed a feed regime, which continuously supplies limiting amounts of inducer in a constant ratio to biomass. For the accurate determination of appropriate amounts of inducer, a time shifted exponential substrate and inducer feed strategy has been developed. The potential of this metabolic and engineering integrated approach was proven in fed-batch cultivation experiments using E. coli HMS174(DE3)(pET11ahSOD) as model system. Furthermore, our strategy enables the use of lactose as inducer, since its consumption can be compensated by appropriate feed profiles. The attained results fully comply with all requirements of industrial large scale cultivation and improve the applicability of strong expression systems. PMID- 14524703 TI - EASE vectors for rapid stable expression of recombinant antibodies. AB - Over the past 10 years, monoclonal antibodies and antibody fragments have become an increasingly important source of therapeutic molecules in the biotechnology industry. Drug development strategies rely on screening large numbers of candidate molecules in search of an optimized drug candidate. This strategy requires efficient production of ten to a few hundred milligrams of candidate molecules for screening in bioassays and animal models. Typically, this amount of recombinant protein expression involves large numbers of transient transfections or cloning of a recombinant cell line. Both of these approaches are time consuming and labor-intensive. In this report, we describe the application of an EASE vector system that is capable of generating stable pools of transfected Chinese hamster ovary cells. These pooled populations of cells produce high quantities of antibody candidates without labor-intensive cloning in a 3-5 week time frame. When an optimal drug candidate has been selected, pools generated with EASE-containing vectors can also be used in subsequent cloning steps to make cell lines with improved expression levels. We demonstrate that EASE increases expression in nonamplified pools in addition to increasing amplification and viability of clonal cell lines generated with the EASE-containing vectors compared with pools and cell lines generated without EASE. PMID- 14524704 TI - Specific oxygen uptake rate variations during batch fermentation of Bacillus thuringiensis subspecies kurstaki HD-1. AB - The specific oxygen uptake rate (q(O)2, respiration rate) of Bacillus thuringiensis subsp. kurstaki HD-1 was very high at inoculation and was found to decrease essentially monotonically throughout both vegetative growth phase and transition phase under different batch culture conditions. Average q(O)2 values decreased from 8-10 mmol/g h at 1 h after inoculation to less than 2 mmol/g h by the time growth ended. The results are shown to be consistent with the few previous reports on q(O)2 in B. thuringiensis in the literature but also novel in that this pattern of monotonic decline has not been described previously. Both pH control and EDTA in low concentration shortened the vegetative growth phase and reduced the 10 h biomass concentration. Using plots of q(O)2 versus specific growth rate, mu, biomass yield based on the oxygen used for growth, was calculated for transition phase to be 0.041-0.047 g/mmol, consistent with literature values. The same plot also showed that the presence of EDTA resulted in an atypical q(O)2-mu trajectory and apparently much higher biomass yield from the oxygen consumed. PMID- 14524705 TI - Metabolic engineering of pentose phosphate pathway in Ralstoniaeutropha for enhanced biosynthesis of poly-beta-hydroxybutyrate. AB - Poly-beta-hydroxybutyrate (PHB) biosynthesis in Ralstonia eutropha from gluconate as a carbon source is carried out through the Entner-Doudoroff (ED) pathway and the pentose-phosphate (PP) pathway generating NADPH and glyceraldehyde-3 phosphate that flows to acetyl-CoA, actively in the unbalanced PHB accumulation phase. The gnd gene encoding 6-phosphogluconate dehydrogenase (6PGDH) and the tktA gene encoding the transketolase (TK) in PP pathway of E. coli were transformed into R. eutropha H16 to modify the metabolic flux of gluconate to the PHB biosynthesis. Over-generated NADPH by the amplified gnd gene tended to depress the cell growth and PHB concentration. Meanwhile, the amplified tktA gene significantly increased both PHB biosynthesis and cell growth as a result of the effective flow of glyceraldehyde-3-phosphate into acetyl-CoA along with the concomitant supplementation of NADPH. The amplified tktA gene also activated the enzyme activities directly associated with PHB biosynthesis. The transformant R. eutropha harboring tktA gene was cultivated using pH-stat-fed-batch to achieve the overproduction of PHB. PMID- 14524706 TI - Altered glucose transport and shikimate pathway product yields in E. coli. AB - Different glucose transport systems are examined for their impact on phosphoenolpyruvate availability as reflected by the yields of 3-dehydroshikimic acid and byproducts 3-deoxy-d-arabino-heptulosonic acid, 3-dehydroquinic acid, and gallic acid synthesized by Escherichia coli from glucose. 3-Dehydroshikimic acid is an advanced shikimate pathway intermediate in the syntheses of a spectrum of commodity, pseudocommodity, and fine chemicals. All constructs carried plasmid aroF(FBR) and tktA inserts encoding, respectively, a feedback-insensitive isozyme of 3-deoxy-d-arabino-heptulosonic acid 7-phosphate synthase and transketolase. Reliance on the native E. coli phosphoenolpyruvate:carbohydrate phosphotransferase system for glucose transport led in 48 h to the synthesis of 3 dehydroshikimic acid (49 g/L) and shikimate pathway byproducts in a total yield of 33% (mol/mol). Use of heterologously expressed Zymomonas mobilis glf-encoded glucose facilitator and glk-encoded glucokinase resulted in the synthesis in 48 h of 3-dehydroshikimic acid (60 g/L) and shikimate pathway byproducts in a total yield of 41% (mol/mol). Recruitment of native E. coli galP-encoded galactose permease for glucose transport required 60 h to synthesize 3-dehydroshikimic acid (60 g/L) and shikimate pathway byproducts in a total yield of 43% (mol/mol). Direct comparison of the impact of altered glucose transport on the yields of shikimate pathway products synthesized by E. coli has been previously hampered by different experimental designs and culturing conditions. In this study, the same product and byproduct mixture synthesized by E. coli constructs derived from the same progenitor strain is used to compare strategies for increasing phosphoenolpyruvate availability. Constructs are cultured under the same set of fermentor-controlled conditions. PMID- 14524707 TI - Chimeric gene library construction by a simple and highly versatile method using recombination-dependent exponential amplification. AB - A simple and efficient method for the construction of chimeric gene libraries termed RDA-PCR (recombination-dependent exponential amplification polymerase chain reaction) was developed by modifying polymerase chain reaction. A chimeric gene library is generated from homologous parental genes with additional primer annealing sequences at their "heads" and "tails". Two primers ("skew primers") are designed to exclusively anneal to either the heads of maternal genes or the tails of paternal genes. During the RDA-PCR, short annealing/extension periods facilitate homologous recombination. The chimeric sequences can be exponentially amplified to form the chimeric gene library, whereas parental sequences without crossovers are not amplified. As a model, we constructed a chimeric gene library of yellow and green fluorescent protein (yfp and gfp, respectively). The crossover point profile of RDA-PCR clones was compared with those obtained by (modified) family shuffling. PCR restriction fragment polymorphism (PCR-RFLP) analysis of the RDA-PCR clones showed a high content of chimeric genes in the library, whereas family shuffling required the modification using skew primers for selective enrichment of chimeric sequences. PCR-RFLP analysis also indicated that the crossover points of RDA-PCR chimeras were distributed over the entire protein-coding region. Moreover, as few as 2 bp of the continual identity of nucleotides were found at the crossover points at high frequency (30% of the tested clones), suggesting that RDA-PCR resulted in a higher diversity in crossover points than family shuffling. PMID- 14524708 TI - Study of process variables in industrial acetic fermentation by a continuous pilot fermentor and response surfaces. AB - The modeling and optimization of industrial processes requires an intensive study of the factors involved. In this work, a continuous pilot system for studying the industrial process of acetic fermentation is developed. A Doehlert design is applied to the five variables involved in the pilot process. This experimental design allows reduction of the experimental burden and the maximum amount of information to be obtained, studying the factors at different levels depending on their significance. The experimental system provides a robust measure of the specific growth rate and the rates of substrates consumption and acetic acid production, related to the flow of effluent stream evaluated in the steady state. The results demonstrate the growth-associated kinetics of substrates and product, and the yield factors are calculated with low values of variances for the coefficients, i.e., within the range 1-11%. The specific growth rate suits the quadratic model proposed. The response surfaces generated by the model are applied to explain the behavior of the bacterial growth and, therefore, the effects of the process variables studied over the acetic acid production. Very low levels of ethanol or oxygen make the acetification rate decrease, and a saturation effect with high levels of ethanol or oxygen is also deduced. The effects of the aeration rate, agitation, and overpressure suggest a kind of inhibition of the acetic acid production caused by the oxygen that has not been practically studied before. The temperature strengthens the inhibitory effect of the ethanol and the oxygen. The conclusions of this work consolidate the structure of a hybrid model for the acetic fermentation. PMID- 14524709 TI - Using computational fluid dynamics software to estimate circulation time distributions in bioreactors. AB - Nonideal mixing in many fermentation processes can lead to concentration gradients in nutrients, oxygen, and pH, among others. These gradients are likely to influence cellular behavior, growth, or yield of the fermentation process. Frequency of exposure to these gradients can be defined by the circulation time distribution (CTD). There are few examples of CTDs in the literature, and experimental determination of CTD is at best a challenging task. The goal in this study was to determine whether computational fluid dynamics (CFD) software (FLUENT 4 and MixSim) could be used to characterize the CTD in a single-impeller mixing tank. To accomplish this, CFD software was used to simulate flow fields in three different mixing tanks by meshing the tanks with a grid of elements and solving the Navier-Stokes equations using the kappa-epsilon turbulence model. Tracer particles were released from a reference zone within the simulated flow fields, particle trajectories were simulated for 30 s, and the time taken for these tracer particles to return to the reference zone was calculated. CTDs determined by experimental measurement, which showed distinct features (log normal, bimodal, and unimodal), were compared with CTDs determined using CFD simulation. Reproducing the signal processing procedures used in each of the experiments, CFD simulations captured the characteristic features of the experimentally measured CTDs. The CFD data suggests new signal processing procedures that predict unimodal CTDs for all three tanks. PMID- 14524710 TI - Cybernetic model predictive control of a continuous bioreactor with cell recycle. AB - The control of poly-beta-hydroxybutyrate (PHB) productivity in a continuous bioreactor with cell recycle is studied by simulation. A cybernetic model of PHB synthesis in Alcaligenes eutrophus is developed. Model parameters are identified using experimental data, and simulation results are presented. The model is interfaced to a multirate model predictive control (MPC) algorithm. PHB productivity and concentration are controlled by manipulating dilution rate and recycle ratio. Unmeasured time varying disturbances are imposed to study regulatory control performance, including unreachable setpoints. With proper controller tuning, the nonlinear MPC algorithm can track productivity and concentration setpoints despite a change in the sign of PHB productivity gain with respect to dilution rate. It is shown that the nonlinear MPC algorithm is able to track the maximum achievable productivity for unreachable setpoints under significant process/model mismatch. The impact of model uncertainty upon controller performance is explored. The multirate MPC algorithm is tested using three controllers employing models that vary in complexity of regulation. It is shown that controller performance deteriorates as a function of decreasing biological complexity. PMID- 14524711 TI - Design of a tubular loop bioreactor for scale-up and scale-down of fermentation processes. AB - Microorganisms traveling through circulation loops in large-scale bioreactors experience variations in their environment such as dissolved oxygen concentration and pH gradients. The same changes are not experienced in small bioreactors, and it is suggested that herein lies one of the major reasons for the problems encountered when translating fermentation data from one scale to another. One approach to study this problem is to look at the circulation loop itself. The present work concerns an attempt to simulate the circulation loops inside stirred tank reactors, using a tubular loop reactor specially constructed for the purpose. The reactor carries a number of ports and probes along its length for the determination of concentration gradients within. The broth is circulated around the loop by the use of peristaltic pumps, and the circulation time (t(c), s) is used as a measure of simulated reactor size. The reactor system has been evaluated using the citric acid fermentation by Aspergillus niger as a test process. Acid production and fungal morphology, in terms of the mean convex perimeter of mycelial clumps quantified by image analysis, were used as the parameters of evaluation for the two systems in comparison. From comparative experiments carried out in 10 and 200 L stirred tank bioreactors, it appears that the loop reactor simulates the corresponding stirred tank representing a valuable tool in scaling up and scaling down of fermentation process. PMID- 14524712 TI - Polymerization of guaiacol and a phenolic beta-O-4-substructure by Trametes hirsuta laccase in the presence of ABTS. AB - The reaction of the monomeric lignin model compound guaiacol and the beta-O-4 type dimer erol (1-(4-hydroxy-3-methoxyphenyl)-2(2-methoxyphenoxy)-propane-1,3 diol with laccase from Trametes hirsuta was studied in the presence of the mediator ABTS (2,2'-azino-di[3ethylbenzothiazoline-6-sulfonic acid]). The product mixtures were analyzed by means of aqueous-phase size exclusion chromatography (SEC) with 50 mM NaOH as eluent. Interestingly, in the laccase-catalyzed reaction with both substrates, the mediator not only functioned as an electron carrier but underwent coupling reactions with the substrate to give polymeric coupling products. The molecular weight of these copolymeric products was significantly higher than the molecular weight of products obtained without ABTS. After ultrafiltration, 33% and 21% of the initially applied ABTS could be found in the polymeric product fraction for the substrates guaiacol and erol, respectively, on the basis of nitrogen analysis. When ABTS was added to substrates after full laccase-catalyzed polymerization, the reaction proceeded toward higher molecular weights. PMID- 14524713 TI - Enzyme distribution derived from macroscopic particle behavior of an industrial immobilized penicillin-G acylase. AB - The macroscopic kinetic behavior of an industrially employed immobilized penicillin-G acylase, called Assemblase, formed the basis for a discussion on some simple intraparticle biocatalytic model distributions. Assemblase catalyzes the synthesis of the widely used semisynthetic antibiotic cephalexin. Despite the obvious advantages of immobilization, less cephalexin and more of the unwanted byproduct d-(-)-phenylglycine are obtained due to diffusional limitations when the immobilized enzyme is employed. To rationally optimize Assemblase, the parameters particle size, enzyme loading, and enzyme distribution, which severely determine the macroscopic particle performance, were studied on the basis of macroscopic observations. Laser diffraction measurements showed that the particle sizes in Assemblase vary as much as 100-fold. The relative and total enzyme loadings in Assemblase and fractions thereof of different sizes were determined by initial-rate d-(-)-phenylglycine amide hydrolysis, cephalexin synthesis experiments, and active-site titration. These experiments revealed that the loading of penicillin-G acylase in Assemblase was inversely correlated with the particle diameter. Apart from enzyme loadings, estimates on the intraparticle enzyme distribution came from cephalexin synthesis experiments, where mass transport limitations were present. Although this method cannot provide the level of detail of specific labeling experiments, it is simple, fast, and cheap. Within the set of simple model predictions, a heterogeneous enzyme distribution with most biocatalyst present in the outer region of the particle (within the outer 100 microm) gave the best description of the observed behavior, although no exact correlation was established. Highly detailed determination of intraparticle enzyme distributions must come from immunolabeling. PMID- 14524714 TI - Supercritical fluid disruption of Ralstonia eutropha for poly(beta hydroxybutyrate) recovery. AB - A new method for disruption of Gram-negative bacterium Ralstonia eutropha by supercritical CO(2) for poly(beta-hydroxybutyrate) (PHB) recovery is proposed. The effects of different parameters such as exposure time, pressure, temperature, volume of methanol as a modifier, and culture history on cell disruption efficiency were investigated using Taguchi's statistical approach to determine optimum conditions. The optimum conditions for cell disruption and PHB recovery were as follows: exposure time, 100 min; pressure, 200 atm; temperature, 40 degrees C; volume of methanol, 0.2 mL. The cell culture time was less significant. At optimum conditions, the maximum efficiency of PHB recovery was found to be 89%. The proposed method is comparable with other recovery methods in terms of the percentage of PHB recovery, while it is environmentally more benign. PMID- 14524715 TI - A predictive aggregate transport model for microfiltration of combined macromolecular solutions and poly-disperse suspensions: model development. AB - A methodology, called the aggregate transport model, is presented that can a priori predict both the pressure-independent permeation flux and yield of target species for the microfiltration of poly-disperse solutions. The model captures the phenomenon of critical shear rate. Beyond the critical shear rate (expressed as a ratio of shear rate to permeation flux), the transmission of proteins drops sharply as a result of cake classification. The widely reported benefits of operating at uniform transmembrane pressure and constant wall concentration follow from this method. The methodology is general in nature and can be used predictively to obtain an optimal balance between flux and yield of target species during the microfiltration of many commercial poly-disperse suspensions. In the accompanying paper we test this model for microfiltration of transgenic whole goat milk. PMID- 14524716 TI - A predictive aggregate transport model for microfiltration of combined macromolecular solutions and poly-disperse suspensions: testing model with transgenic goat milk. AB - To meet the technical challenge of recovering human IgG fusion protein from transgenic whole goat milk at reasonable cost with high purity and yield, a predictive aggregate transport model for microfiltration has been developed (Baruah and Belfort, 2003). Here, to test the model's predictability of permeate flux and mass transport, a comprehensive series of experiments with varying wall shear rate, feed temperature, feed concentration, and module design are presented. A very good fit was obtained between the model predictions and measurements for a wide variety of experimental conditions. For microfiltration module design comparison, a linear hollow fiber module (representing current commercial technologies) gave lower permeation flux and higher yield than a helical hollow fiber module (representing the latest self-cleaning methodology). These results are easily explained with the model that is now being used to define operating conditions for maximizing performance. The procedure described by the model is generalizable and can be used to obtain optimal filtration performance for applications other than milk. PMID- 14524718 TI - Effective encapsulation of proteins into size-controlled phospholipid vesicles using freeze-thawing and extrusion. AB - We are aiming to improve the encapsulation efficiency of proteins in a size regulated phospholipid vesicle using an extrusion method. Mixed lipids (1,2 dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC), cholesterol, 1,5-dipalmitoyl l-glutamate-N-succinic acid (DPEA), and 1,2-distearoyl-sn-glycero-3 phosphoethanolamine-N-[monomethoxy poly(ethylene glycol) (5,000)] (PEG-DSPE) at a molar ratio of 5, 5, 1, and 0.033 were hydrated with a NaOH solution (7.6 mM) to obtain a polydispersed multilamellar vesicle dispersion (50 nm to 30 microm diameter). The polydispersed vesicles were converted to smaller vesicles having an average diameter of ca. 500 nm with a relatively narrow size distribution by freeze-thawing at a lipid concentration of 2 g dL(-)(1) and cooling rate of -140 degrees C min(-1). The lyophilized powder of the freeze-thawed vesicles was rehydrated into a concentrated protein solution (carbonyl hemoglobin solution, 40 g dL(-1)) and retained the size and size distribution of the original vesicles. The resulting vesicle dispersion smoothly permeated through the membrane filters during extrusion. The average permeation rate of the freeze-thawed vesicles was ca. 30 times faster than that of simple hydrated vesicles. During the extrusion process, proteins were encapsulated into the reconstructed vesicles with a diameter of 250 +/- 20 nm. PMID- 14524717 TI - Optimization of an osmotic shock procedure for isolation of a protein product expressed in E. coli. AB - Small-scale experiments were performed to evaluate the impact of several processing variables on recovery by osmotic shock of a protein product expressed in E. coli utilizing feed material manufactured at large scale. Equilibration time, type and concentration of osmotic supplements, and pH of the osmotic shock buffer were found to have significant impact on the osmotic shock recovery. Improved recovery was documented under conditions of increased equilibration time before the osmotic shock and increased Tris-HCl concentration in the osmotic shock buffer, as well as initial dispersal of cells into the osmotic shock buffer. Recovery of up to 65% was achieved under the optimized conditions. PMID- 14524720 TI - Micropatterned substrates for the growth of functional neuronal networks of defined geometry. AB - The in vitro assembly of neuronal networks with control over cell position and connectivity is a fascinating approach not only for topics in basic neuroscience research but also in diverse applications such as biosensors and tissue engineering. We grew rat embryonic cortical neurons on patterned substrates created by microcontact printing. Polystyrene was used as a cell repellent background, onto which a grid pattern of physiological proteins was applied. We printed laminin and a mixture of extracellular matrix proteins and additionally both systems mixed with polylysine. Attachment of cells to the pattern with high fidelity as well as the formation of chemical synapses between neighboring cells on the pattern could be observed in all four cases, but cell attachment was strongly increased on samples containing polylysine. Neurons grown on patterned substrates had a membrane capacity smaller than that of neurons on homogeneously coated controls, which we attributed to the geometrical restrictions, but did not differ either in resting membrane potential or in the quality of synapses they formed. We therefore believe that the cells attach and differentiate normally on the pattern and form functional, mature synapses following the predefined geometry. PMID- 14524719 TI - Mathematical and experimental analyses of antibody transport in hollow-fiber based specific antibody filters. AB - We are developing hollow fiber-based specific antibody filters (SAFs) that selectively remove antibodies of a given specificity directly from whole blood, without separation of the plasma and cellular blood components and with minimal removal of plasma proteins other than the targeted pathogenic antibodies. A principal goal of our research is to identify the primary mechanisms that control antibody transport within the SAF and to use this information to guide the choice of design and operational parameters that maximize the SAF-based antibody removal rate. In this study, we formulated a simple mathematical model of SAF-based antibody removal and performed in vitro antibody removal experiments to test key predictions of the model. Our model revealed three antibody transport regimes, defined by the magnitude of the Damkohler number Da (characteristic antibody binding rate/characteristic antibody diffusion rate): reaction-limited (Da /= 10). For a given SAF geometry, blood flow rate, and antibody diffusivity, the highest antibody removal rate was predicted for diffusion-limited antibody transport. Additionally, for diffusion-limited antibody transport the predicted antibody removal rate was independent of the antibody-binding rate and hence was the same for any antibody-antigen system and for any patient within one antibody-antigen system. Using SAF prototypes containing immobilized bovine serum albumin (BSA), we measured anti-BSA removal rates consistent with transport in the intermediate regime (Da approximately 3). We concluded that initial SAF development work should focus on achieving diffusion-limited antibody transport by maximizing the SAF antibody-binding capacity (hence maximizing the characteristic antibody binding rate). If diffusion-limited antibody transport is achieved, the antibody removal rate may be raised further by increasing the number and length of the SAF fibers and by increasing the blood flow rate through the SAF. PMID- 14524721 TI - Attachment of marine sponge cells of Hymeniacidon perleve on microcarriers. AB - Toward the development of an in vitro cultivation of marine sponge cells for sustainable production of bioactive metabolites, the attachment characteristics of marine sponge cells of Hymeniacidon perleve on three types of microcarriers, Hillex, Cytodex 3, and glass beads, were studied. Mixed cell population and enriched cell fractions of specific cell types by Ficoll gradient centrifugation (6%/8%/15%/20%) were also assessed. Cell attachment ratio (defined as the ratio of cells attached on microcarrier to the total number of cells in the culture) on glass beads is much higher than that on Cytodex 3 and Hillex for both mixed cell population and cell fraction at Ficoll 15-20% interface. The highest attachment ratio of 41% was obtained for the cell fraction at Ficoll 15-20% interface on glass beads, which was significantly higher than that of a mixed cell population (18%). The attachment kinetics on glass beads indicated that the attachment was completed within 1 h. Cell attachment ratio decreases with increase in cell-to microcarrier ratio (3-30 cells/bead) and pH (7.6-9.0). The addition of serum and BSA (bovine serum albumin) reduced the cell attachment on glass beads. PMID- 14524722 TI - Effects of three-dimensional culturing on osteosarcoma cells grown in a fibrous matrix: analyses of cell morphology, cell cycle, and apoptosis. AB - Osteosarcoma cells were cultured in stirred tank bioreactors with either a fibrous matrix or nonporous microcarriers to study the environmental effects on cell growth, morphology, cell cycle, and apoptosis. Cell cycle and apoptosis were analyzed using flow cytometry and visualized using confocal laser scanning microscopy and fluorescence microscopy. The three-dimensional (3-D) fibrous culture had better cell growth and higher metabolic rates than the two dimensional (2-D) microcarrier culture because cells in the fibrous matrix were protected from shear stress and had lower apoptosis and cell death even under suboptimal conditions (e.g., nutrient depletion). The polyester fibrous matrix used in this study also exhibited the capability of selectively retaining viable and nonapoptotic cells and disposing apoptotic and nonviable cells. Consequently, very few apoptotic cells were found in the fibrous matrix even in the long-term (1 month) T-flask culture. In the continuous culture with packed fibrous matrixes for cell support, most cells were arrested in the G1/G0 phase after 4 days. Decreasing the dissolved oxygen level from 60 to 10% air saturation did not significantly change cell cycle and apoptosis, which remained low at approximately 15%. These results could explain why the fibrous bed bioreactor had good long-term stability and was advantageous for production of non-growth associated proteins by animal cell cultures. PMID- 14524723 TI - Predicted unfolding order of the 13 alpha-helices in the catalytic domain of glucoamylase from Aspergillus awamori var. X100 by molecular dynamics simulations. AB - The unfolding mechanism of the 13 alpha-helices in the catalytic domain of Aspergillus awamori var. X100 glucoamylase was investigated by 200 ps molecular dynamics simulations in explicit water with temperature jump technique. Rather than a simultaneous event, the unfolding of these 13 alpha-helices followed a random ordered mechanism as alpha8-->alpha1-->alpha11-->alpha7-->alpha10-->alpha3 ->alpha12-->alpha13-->alpha4-->alpha5-->alpha9-->alpha6-->alpha2. No significant relationships were found between the unfolding order and the length and the hydrophobicity of the helix. alpha-Helix 8 located in the inner region of the catalytic domain was predicted to be the first helix to unfold, indicating that the destruction of the secondary structure motif was initiated from the inner region of the catalytic domain. The dynamic behavior of these alpha-helices induced by increased kinetic energy during the unfolding process is considered to be similar to the expansion and compression of a series of springs under the influence of mechanical stress. PMID- 14524724 TI - Data-based modeling and analysis of bioprocesses: some real experiences. AB - Data-generated models find numerous applications in areas where the speed of collection and logging of data surpasses the ability to analyze it. This work is meant to addresses some of the challenges and difficulties encountered in the practical application of these methods in an industrial setting and, more specifically, in the bioprocess industry. Neural network and principal component models are the two topics that are covered in detail in this paper. A review of these modeling technologies as applied to bioprocessing is provided, and four original case studies using industrial fermentation data are presented that utilize these models in the context of prediction and monitoring of bioprocess performance. PMID- 14524725 TI - Effect of oxygen delignification operating parameters on downstream enzymatic hydrolysis of softwood substrates. AB - Enhanced oxygen delignification of softwood pulp samples (taken upstream and downstream of a commercial oxygen delignification unit) improved the initial rate of enzymatic saccharification and overall yield of monomeric sugars by 62-82% and 76-80%, respectively. Laboratory-scale experiments were used to examine the effect of a broad range of operating parameters (temperature, time, caustic concentration, and oxygen partial pressure) on the effectiveness of oxygen delignification. Using empirical models, kappa number (residual lignin content) was found to effectively predict final conversion to monomeric sugars. Application of oxygen delignification to sulfite mill knots resulted in smaller (20-25%) reduction in lignin content. However, using a combination of oxygen delignification and particle size reduction, up to 80% of the carbohydrate in the reject knots could be converted to fermentable sugars. PMID- 14524726 TI - Cell surface display of organophosphorus hydrolase in Pseudomonas putida using an ice-nucleation protein anchor. AB - A surface anchor system derived from the ice-nucleation protein (INP) from Pseudomonas syringe was used to localize organophosphorus hydrolase (OPH) onto the surface of Pseudomonas putida KT2440. Cells harboring the shuttle vector pPNCO33 coding for the INP-OPH fusion were capable of targeting OPH onto the cell surface as demonstrated by whole cell ELISA. The whole cell activity of P. putida KT2440 was shown to be 10 times higher than those of previous efforts expressing the same fusion protein in Escherichia coli. The capability of expressing enzymes on the surface of a robust and environmentally benign P. putida KT2440 should open up new avenues for a wide range of applications such as in situ bioremediation. PMID- 14524727 TI - Novel fermentation strategy for enhancing glycerol production by Candida krusei. AB - During the later stage of glycerol production by fermentation of Candida krusei, glycerol consumption by the strain was observed, although there was residual sugar in the medium. To enhance the final glycerol accumulation, a new fermentation strategy was developed by maintaining high activities of glycerol synthetic enzymes (i.e., glycerol-3-phosphate dehydrogenase (ctGPD) and glycerol 3-phosphatase (GPP)) for a relatively long period while conducting oxygen limitation at a later stage to inhibit the increase of another enzyme activity related to glycerol degradation (i.e., mitochondrial glycerol-3-phosphate dehydrogenase (mtGPD)). With oxygen limitation performed from 88 h, when ctGPD and GPP activities were already at a low level while mtGPD activity was increasing, the glycerol dissimilation was efficiently reduced. The final glycerol concentration reached 55.6 g/L, which was about 18% (96 h) and 30% (104 h) higher than control, and its productivity increased to 0.54 g/(L h). The proposed strategy based on cell physiology was proved useful to the glycerol fermentation process. PMID- 14524728 TI - Improved mammalian expression systems by manipulating transcriptional termination regions. AB - Here we show that the transcriptional terminator element of human gastrin gene, which is the only element characterized to date in terms of its function in transcriptional termination, increases the transient expression levels of recombinant proteins. The expression of the beta-galactosidase gene was enhanced 3-4-fold in HeLa cells by inserting the terminator element of human gastrin gene at the 3'-side of the SV40 polyadenylation signal/cleavage site of the control vector (pSV-beta-gal). This effect of the terminator element is orientation dependent but not cell-specific since a similar enhancement of beta-galactosidase gene expression was detected in COS.M6 and CHO DG44 cells. The increased level of beta-galactosidase gene expression by the transcriptional terminator element of human gastrin gene turned out to arise from elevated cellular mRNA levels, suggesting that the terminator element stabilizes mRNA by enhancing proper 3'-end processing of mRNA. PMID- 14524729 TI - Size and stability of reconstituted sesame oil bodies. AB - Oil bodies of sesame seeds comprise a triacylglycerol matrix, which is surrounded by a monolayer of phospholipids embedded with unique proteins, mainly structural proteins termed oleosins. Artificial oil bodies were successfully reconstituted with various compositions of triacylglycerols, phospholipids, and oil-body proteins. The sizes of reconstituted oil bodies displayed a normal distribution with an average size proportional to the ratio of triacylglycerols to oil-body proteins. Both thermostability and structural stability of reconstituted oil bodies decreased as their sizes increased, and vice versa. Proteinase K digestion indicated that oleosins anchored both native and reconstituted oil bodies via their central hydrophobic domains. The stability of reconstituted oil bodies, as well as the purified ones from sesame seeds, could be substantially enhanced after their surface proteins were cross-linked by glutaraldehyde or genipin. PMID- 14524730 TI - Pharmacological management of first-episode schizophrenia and related nonaffective psychoses. AB - Schizophrenia is a severe mental illness characterised by abnormalities of thought and perception that affects 1-2% of the population. Patients who experience a first episode of schizophrenia should be treated early and optimally with antipsychotic agents to lessen the morbidity of the initial episode and possibly improve the course of the illness. Positive psychotic symptoms remit in the majority of patients who are treated with adequate trials of antipsychotic medications, but most relapse within 1 year. Non-adherence is strongly related to the likelihood of recurrence of symptoms. Innovative programmes that integrate early intervention, psychosocial treatments and atypical antipsychotic pharmacotherapy show promise in improving outcomes. The available research supports the use of antipsychotic medications early in the first-episode of schizophrenia and for at least 1 year after remission of positive symptoms. Antidepressants, benzodiazepines and mood stabilisers have roles in the acute and maintenance phases of treatment for some patients. Atypical antipsychotics represent a great advance in the treatment of first-episode schizophrenia with strong evidence for greater tolerability with equal or better therapeutic efficacy. Future research will further define their roles in treatment and hopefully identify targets for prevention of first-episode schizophrenia. PMID- 14524732 TI - Improving health-related quality of life in gastro-oesophageal reflux disease. AB - Gastro-oesophageal reflux disease (GORD) is a common, chronic disorder that can progress to erosive or ulcerative oesophagitis and other complications. Uncomplicated GORD can be defined using criteria that combine symptom assessment and the impact of symptoms on well being or quality of life. The extent of impairment of health-related quality of life (HRQL) in GORD patients has been evaluated using generic HRQL questionnaires, GORD-specific quality of life questionnaires or a combination of the two types of instruments. The impact of GORD on HRQL in affected individuals is proportional to the frequency and severity of heartburn, and is greater than that associated with many other chronic diseases. In the treatment of GORD, the goal should be complete symptom resolution as this has been shown to be associated with a clinically significant improvement in HRQL. Both drug treatment and surgical intervention have been successful in improving HRQL of patients with GORD when complete symptom resolution can be attained. PMID- 14524734 TI - Oral fludarabine: a viewpoint by Dieter Huhn. PMID- 14524731 TI - Newer formulations of the triptans: advances in migraine management. AB - Migraine is a common, frequently incapacitating, headache disorder that imposes a substantial burden on both the individual patient and society. The last two decades have witnessed an explosion in our understanding of the pathophysiology of migraine, and in our development of an efficacious and diverse therapeutic armamentarium. There are several routes of drug administration available to patients with migraine. All the serotonin 5-HT(1B/1D) receptor agonists (triptans) are available as oral tablets (sumatriptan, rizatriptan, zolmitriptan, naratriptan, almotriptan, frovatriptan and eletriptan). Only sumatriptan is available as a subcutaneous injection. Some triptans are also available via newer routes of administration, including orally disintegrating tablets (rizatriptan and zolmitriptan), rectal suppositories (sumatriptan) and intranasal sprays (sumatriptan and zolmitriptan). Oral disintegrating tablets and other non-oral triptan routes (subcutaneous, intranasal, rectal) are a useful alternative to conventional oral tablets for patients who have difficulty swallowing pills or prefer not to do so, and for patients whose nausea and/or vomiting precludes swallowing tablets and/or makes the likelihood of complete absorption unpredictable. This is important because epidemiological studies in migraine reveal that the vast majority of patients (>90%) have experienced nausea during a migraine attack and more than 50% have nausea with the majority of attacks. Similarly, most (almost 70%) have vomited at some time during an attack and of these patients, almost one-third vomit in the majority of attacks. The newer formulations, rapidly dissolving tablets and intranasal sprays, afford patients the opportunity to use abortive therapy without the need for liquids, at anytime and anywhere, at the onset of a migraine attack. Furthermore, the intranasal sprays are absorbed rapidly and have a prompt onset of action allowing for significant pain free rates versus placebo as early as 15 minutes post administration. The ability to administer treatment early in a migraine attack and have a rapid onset of action is particularly important in acute migraine treatment in order to prevent the development of central sensitisation. While many patients and physicians choose conventional oral tablets because of familiarity and ease of administration, the newer formulations, oral disintegrating tablets and intranasal sprays, should be given consideration as first-line agents in selected patients. PMID- 14524735 TI - Oral fludarabine: a viewpoint by David Oscier. PMID- 14524733 TI - Oral fludarabine. AB - Fludarabine is an antimetabolite antineoplastic agent used in the treatment of various haematological malignancies, particularly B-cell chronic lymphocytic leukaemia (CLL). An oral formulation of fludarabine has recently become available in the majority of European countries for the treatment of patients with relapsed or refractory B-cell CLL after initial treatment with an alkylating agent-based regimen. It is the first oral formulation of a purine analogue available for clinical use in B-cell CLL. Pharmacokinetic studies evaluating the bioavailability of oral fludarabine indicate that an oral dose of 40 mg/m2/day would provide similar systemic drug exposure to the standard intravenous dose of 25 mg/m2/day. A phase II study evaluated the clinical efficacy of six to eight cycles of oral fludarabine 40 mg/m2/day for 5 days of each 28-day cycle in 78 patients with previously treated B-cell CLL. Depending on the criteria used, the overall response rate was 46.2% (20.5% complete response [CR], 25.6% partial response [PR]) or 51.3% (17.9% CR, 33.3% PR). These results were similar to the 48% overall response rate reported in a similar historical control group treated with intravenous fludarabine. Myelosuppression (WHO grade 3 or 4) was the most frequently reported adverse effect with oral fludarabine therapy. Other common adverse effects included infection and gastrointestinal disturbances, although these were usually of mild to moderate severity (WHO grade 1 or 2). Overall, the tolerability profile of oral fludarabine is similar to that of the intravenous formulation. PMID- 14524736 TI - Oral fludarabine: a viewpoint by Bruce D. Cheson. PMID- 14524738 TI - Bemiparin: a review of its use in the prevention of venous thromboembolism and treatment of deep vein thrombosis. AB - Bemiparin (bemiparin sodium; Hibor, Ivor, Zibor, Badyket) is a low molecular weight heparin (LMWH) with a lower mean molecular weight (3600 D) and a higher anti-Xa/IIa ratio (8:1) than other LMWHs. Bemiparin was effective as thromboprophylaxis in surgical patients in well controlled clinical trials. No cases of venous thromboembolism (VTE) were reported in low- to moderate-risk patients receiving prophylaxis with bemiparin 2500 anti-Xa IU/day for 7 days or unfractionated heparin (UFH) 5000 anti-Xa IU twice daily for 7 days. In high-risk patients, bemiparin 3500 anti-Xa IU/day for > or =8 days was more effective than UFH 5000 anti-Xa IU twice daily for > or =8 days in the prevention of VTE in patients undergoing total hip replacement. Postoperative bemiparin 3500 anti-Xa IU/day for 10 days was as effective as enoxaparin 4000 anti-Xa IU/day for 10 days commenced 12 hours before surgery in high-risk patients undergoing total knee replacement. As a short-term treatment for acute established deep vein thrombosis (DVT), bemiparin 5000-10 000 anti-Xa IU/day (dependent on bodyweight) for 7 or 10 days was more effective than intravenous UFH (5000 anti-Xa IU bolus followed by 30,000 or 40,000 anti-Xa IU/day for 7 days) in reducing thrombus size from baseline. Bemiparin 3500 anti-Xa IU/day was also as effective as oral warfarin (10 mg/day for the first 3 days, then adjusted to achieve an international normalised ratio between 2 and 3) for the long-term (12 weeks) treatment of DVT, although data are limited. Subcutaneous bemiparin was generally well tolerated. The most commonly reported adverse events in clinical trials were postoperative bleeding complications (similar incidence to that with UFH or enoxaparin in high risk patients, lower incidence in low- to moderate-risk patients). CONCLUSIONS: Bemiparin is a new LMWH which has shown efficacy in a small number of well controlled trials in the prevention of postoperative VTE in low- to moderate- and high-risk patients and in the treatment of established DVT. It can be initiated pre- or post-operatively, whereas recommendations for other LMWHs in Europe primarily involve preoperative initiation. Additional comparative studies would be beneficial in determining the overall place of bemiparin, particularly with respect to the relative incidence of bleeding complications. In the meantime, available data suggest that bemiparin is an effective and useful addition to the available range of LMWHs for the prevention of VTE and treatment of DVT. PMID- 14524739 TI - Persistent global power fluctuations near a dynamic transition in electroconvection. AB - This is a study of the global fluctuations in power injection and light transmission through a liquid crystal just above the onset of electroconvection. The source of the fluctuations is identified as the creation and annihilation of defects. They are spatially uncorrelated and yet temporally correlated. The temporal correlation is seen to persist for extremely long times. There seems to be an especially close relation between defect creation or annihilation in electroconvection and thermal plumes in Rayleigh-Benard convection. PMID- 14524740 TI - Behavior of fractional diffusion at the origin. AB - The present work discusses the fractional diffusion equation based on the Riemann Liouville fractional time derivatives. It was shown that the normalization conservation constraint leads to the divergency of diffusive agent concentration at the origin. This divergency implies an external source of the diffusive agent at r-->0. Thus, the Riemann-Liouville fractional time derivative implies a loss of diffusive agent mass, which is compensated for by the source of this agent at the origin. In contrast, the absence of the normalization conservation constraint does not lead to any divergences in the limit r-->0 and at the same time provides the decay of normalization. PMID- 14524741 TI - Jamming during the discharge of grains from a silo described as a percolating transition. AB - We have looked into an experiment that has been termed the "canonical example" of jamming: granular material, clogging the outlet of a container as it is discharged by gravity. We present quantitative data of such an experiment. The experimental control parameter is the ratio between the radius of the orifice and the radius of the beads. As this parameter is increased, the jamming probability decreases. However, in the range of parameters explored, no evidence of criticality--in the sense of a jamming probability that becomes infinitely small for a finite radius--has been found. We draw instead a comparison with a simple model that captures the main features of the phenomenon, namely, percolation in one dimension. The model gives indeed a phase transition, albeit a special one. PMID- 14524737 TI - Lacidipine: a review of its use in the management of hypertension. AB - Lacidipine (Caldine, Lacimen, Lacipil, Midotens, Motens) is a once-daily, orally administered, lipophilic dihydropyridine calcium antagonist with an intrinsically slow onset of activity, resulting in a lack of reflex tachycardia. It has a long duration of action and a high degree of vascular selectivity. In addition to calcium channel-modulated vasodilation, lacidipine displays antioxidant activity greater than that of other dihydropyridine calcium antagonists. In randomised, well-controlled trials, lacidipine 2-6 mg orally once daily had antihypertensive efficacy similar to that of other long-acting dihydropyridine calcium antagonists, thiazide diuretics, atenolol (a beta-blocker) and enalapril (an ACE inhibitor). Lacidipine was effective in elderly patients (including those with isolated systolic hypertension), African Nigerian patients and patients with concurrent type 2 diabetes mellitus. During long-term treatment for 4 or 5 years in patients with isolated systolic hypertension or essential hypertension, the incidence of cardiovascular events and mortality with lacidipine was similar to that with chlorthalidone or atenolol. The European Lacidipine Study on Atherosclerosis (ELSA), in which 2334 patients with hypertension were randomised to 4 years of therapy with lacidipine 4-6 mg/day or the beta-blocker atenolol 50 100 mg/day, demonstrated significantly lower atherosclerotic progression and plaque formation with lacidipine compared with atenolol in patients completing the full 4 years of the study. Between-group differences in favour of lacidipine for the primary efficacy variable (mean change in carotid artery intima-media thickness) did not reach statistical significance in the intent-to-treat population. The tolerability profile of lacidipine (headache, flushing, pedal oedema, dizziness and palpitations) is similar to that of other dihydropyridine calcium antagonists, but with a lower incidence of peripheral oedema. Data from the ELSA study suggest that the incidence of serious adverse events during long term lacidipine therapy is similar to that with atenolol. CONCLUSION: Lacidipine is an effective, well tolerated, once-daily, oral antihypertensive agent that can be used in a wide variety of patients. As with other members of its class, lacidipine has shown potentially beneficial antiatherosclerotic effects, although definitive data with respect to possible superiority over other drug classes are still required. Therefore, lacidipine is an attractive therapy for the long-term management of essential hypertension. PMID- 14524742 TI - Hydrodynamic modes for granular gases. AB - The eigenfunctions and eigenvalues of the linearized Boltzmann equation for inelastic hard spheres (d=3) or disks (d=2) corresponding to d+2 hydrodynamic modes are calculated in the long wavelength limit for a granular gas. The transport coefficients are identified and found to agree with those from the Chapman-Enskog solution. The dominance of hydrodynamic modes at long times and long wavelengths is studied via an exactly solvable kinetic model. A collisional continuum is bounded away from the hydrodynamic spectrum, assuring a hydrodynamic description at long times. The bound is closely related to the power law decay of the velocity distribution in the reference homogeneous cooling state. PMID- 14524743 TI - Prisoners' dilemma in real-world acquaintance networks: spikes and quasiequilibria induced by the interplay between structure and dynamics. AB - We study Nowak and May's spatial prisoners' dilemma game driven by mutations (random choices of suboptimal strategies) on empirical social networks. The time evolution of the cooperation level is highly complex containing spikes and steps between quasistable levels. A statistical characterization of the quasistable states and a study of the mechanisms behind the steps are given. We argue that the crucial structural ingredients causing the observed behavior is an inhomogeneous degree distribution and that the connections within vertices of highest degree are rather sparse. Based on these observations we construct model networks with a similar complex time evolution of the cooperation level. PMID- 14524744 TI - Geometry of escort distributions. AB - Given an original distribution, its statistical and probabilistic attributes may be scanned using the associated escort distribution introduced by Beck and Schlogl and employed in the formulation of nonextensive statistical mechanics. Here, the geometric structure of the one-parameter family of the escort distributions is studied based on the Kullback-Leibler divergence and the relevant Fisher metric. It is shown that the Fisher metric is given in terms of the generalized bit variance, which measures fluctuations of the crowding index of a multifractal. The Cramer-Rao inequality leads to a fundamental limit for the precision of the statistical estimate of the order of the escort distribution. We also show quantitatively that it is inappropriate to use the original distribution instead of the escort distribution for calculating the expectation values of physical quantities in nonextensive statistical mechanics. PMID- 14524745 TI - Persistent random walk in a honeycomb structure: light transport in foams. AB - We study light transport in a honeycomb structure as the simplest two-dimensional model foam. We apply geometrical optics to set up a persistent random walk for the photons. For three special injection angles of 30 degrees, 60 degrees, and 90 degrees relative to a hexagon's edge, we are able to demonstrate by analytical means the diffusive behavior of the photons and to derive their diffusion constants in terms of intensity reflectance, edge length, and velocity of light. Numerical simulations reveal an interesting dependence of the diffusion constant on the injection angle in contrast to the usual assumption that in the diffusive limit the photon has no memory for its initial conditions. Furthermore, for injection angles close to 30 degrees, the diffusion constant does not converge to the value at 30 degrees. We explain this observation in terms of a two-state model. PMID- 14524746 TI - Quantum diffusion on a cyclic one-dimensional lattice. AB - The quantum diffusion of a particle in an initially localized state on a cyclic lattice with N sites is studied. Diffusion and reconstruction time are calculated. Strong differences are found for even or odd number of sites and the limit N--> infinity is studied. The predictions of the model could be tested with microtechnology and nanotechnology devices. PMID- 14524747 TI - Exchange-driven growth. AB - We study a class of growth processes in which clusters evolve via exchange of particles. We show that depending on the rate of exchange there are three possibilities: (I) Growth-clusters grow indefinitely, (II) gelation-all mass is transformed into an infinite gel in a finite time, and (III) instant gelation. In regimes I and II, the cluster size distribution attains a self-similar form. The large size tail of the scaling distribution is Phi(x) approximately exp(-x(2 nu)), where nu is a homogeneity degree of the rate of exchange. At the borderline case nu=2, the distribution exhibits a generic algebraic tail, Phi(x) approximately x(-5). In regime III, the gel nucleates immediately and consumes the entire system. For finite systems, the gelation time vanishes logarithmically, T approximately [lnN](-(nu-2)), in the large system size limit N -> infinity. The theory is applied to coarsening in the infinite range Ising Kawasaki model and in electrostatically driven granular layers. PMID- 14524748 TI - Sensitivity of explosion to departure from partial equilibrium. AB - We study a thermochemical gaseous system in the vicinity of the bifurcation related to the emergence of bistability. Corrections to the standard deterministic dynamics induced by the perturbation of the particle velocity distribution are obtained from the solution of the Boltzmann equation. Using these results, analytical expressions including the nonequilibrium effects are derived for the ignition time in the explosive regime and mean first passage time in the bistable regime. It is demonstrated that a departure from partial equilibrium can shift the bifurcation point. The system which was bistable according to the standard deterministic approach, can become monostable and explosive in the presence of nonequilibrium effects. Even when the system remains in the bistable regime, the mean first passage time can be changed by several orders of magnitude. In the monostable domain, the ignition time can be about ten times smaller than the unperturbed value. These analytical predictions agree well with the results of the microscopic simulations of the dilute gas system. PMID- 14524749 TI - Attempt to distinguish electric signals of a dichotomous nature. AB - Three types of electric signals were analyzed: Ion current fluctuations in membrane channels (ICFMC), Seismic electric signals activities (SES), and "artificial" noises (AN). The wavelet transform, when applied to the conventional time domain, does not allow a classification of these signals, but does so in the "natural" time domain. A classification also becomes possible, if we study -(q) versus q, where chi stands for the "natural" time. For q values approximately between 1 and 2 the signals are classified and ICFMC lies between the other two types. For q=1, the "entropy" S identical with ln of ICFMC almost equals that of a "uniform" distribution, while the AN and SES have larger and smaller S values, respectively. The recent [P. Varotsos, N. Sarlis, and E. Skordas, Phys. Rev. E 67, 021109 (2003)] finding that, in short time scales, both SES and AN (which are shown to be non-Markovian) result in comparable detrended fluctuation analysis exponents alpha in (1.0,1.5) is revisited. Even a Markovian dichotomous time series, in short time scales, leads to similar alpha exponents. PMID- 14524750 TI - Suprathreshold stochastic resonance and noise-enhanced Fisher information in arrays of threshold devices. AB - We analyze the parametric estimation that can be performed on a signal buried in noise based on the parsimonious representation provided by a parallel array of threshold devices. The Fisher information contained in the array output about the input parameter is used as the measure of performance in the estimation task. For estimation on a suprathreshold input signal, we establish that enhancement of the Fisher information can be obtained by addition of independent noises to the thresholds in the array. Similar improvement by noise is also shown to be possible for the estimation error of the maximum likelihood estimator. These results extend the applicability of the recently introduced nonlinear phenomenon of suprathreshold stochastic resonance. PMID- 14524751 TI - Fractional dynamics and nonlinear harmonic responses in dielectric relaxation of disordered liquids. AB - The problem of the nonlinear dielectric response due to the application of a strong electric field is reconsidered in the context of fractional kinetic equations. To accomplish that, we start from a fractional noninertial Fokker Planck equation and restrict ourselves to the case of anomalous subdiffusive processes characterized by the critical exponent alpha ranging from 0 to 1, the limit of normal diffusion. In particular, we evaluate the first- and third-order nonlinear harmonic components of the electric polarization in the case of either a pure ac field or a strong dc bias field superimposed on a weak ac field. The stationary regime is therefore calculated from an infinite set of differential recurrence relations by using a perturbation method. The results so obtained are illustrated by three-dimensional dispersion and absorption plots in order to show the influence of alpha. Cole-Cole diagrams are also presented, allowing one to see that the arcs become more and more flattened as alpha-->0, and corresponding to a broadening of the absorption peaks as effectively observed in complex liquids. The theoretical model is supported by comparison with experimental data of the third-order nonlinear dielectric permittivity of a ferroelectric liquid crystal. PMID- 14524752 TI - Reexamination of string phase and shear thickening in simple fluids. AB - In 1984, Erpenbeck observed a shear-induced alignment of particles into strings in nonequilibrium molecular dynamics simulations of hard spheres. Since then, it has remained unclear if this effect was genuine or if it arose from the use of a thermostat which assumed an incorrect form for the velocity profile. All studies performed up to now have focused on improving the accuracy with which the velocity profile is determined. We propose here a radically different approach: we apply a recently developed configurational expression for the temperature. This expression does not require any knowledge of the streaming velocity profile. Using a configurational thermostat, we show that the string phase is an artifact and we observe a shear-thickening regime, as seen in experiments on concentrated "hard-sphere"-like colloidal dispersions. PMID- 14524753 TI - Transport properties of ferrofluids. AB - Long-time self-diffusion coefficients of ferrofluid suspensions are derived from an effective Langevin equation approach. The dependences of these transport properties on the volume fraction of particles and the strength of interparticle interaction are investigated. Strong reduction of the rotational and the translational Brownian motion of the particles is manifested upon increase of particle-particle interaction or ferrofluid concentration. PMID- 14524754 TI - Field-induced pseudocrystalline ordering in concentrated ferrofluids. AB - Concentrated surfactant stabilized cobalt ferrofluids up to 6 vol % Co have been studied by small-angle scattering using polarized neutrons and synchrotron x rays. The combination of these techniques allowed the magnetic and nuclear form factors to be reliably separated from the structure factors. Above 1 vol % Co, inter particle interactions are induced by an applied external magnetic field that gives rise to pseudocrystalline ordering of cobalt core-shell particles. Particles are arranged in hexagonal planes, with the magnetic moments aligned parallel to the [110] direction. Two types of equivalent textures were found to be present simultaneously, corresponding to a stacking of the hexagonal planes in horizontal and vertical direction. The in-plane nearest-neighbor distance is almost independent of the concentration and temperatures, whereas the distance between the neighboring planes, c, strongly varies from sample to sample. In addition, segments of chains of particles with parallel moments are aligned along the magnetic field and frozen-in when the carrier liquid is solidified. The field induced pseudocrystalline lamellar hexagonal particle arrangement, observed experimentally in colloidal magnetic liquids, confirms predictions from molecular dynamics and Monte Carlo simulations. PMID- 14524755 TI - Transport properties of dense fluid argon. AB - We calculate using molecular dynamics simulations the transport properties of realistically modeled fluid argon at pressures up to approximately 50 GPa and temperatures up to 3000 K. In this context, we provide a critique of some newer theoretical predictions for the diffusion coefficients of liquids and a discussion of the Enskog theory relevance under two different adaptations: modified Enskog theory and effective diameter Enskog theory. We also analyze a number of experimental data for the thermal conductivity of monoatomic and small diatomic dense fluids. PMID- 14524756 TI - Relative pair dynamics in simple supercooled liquids: longitudinal contributions. AB - The pair dynamics of simulated argon samples is investigated at the melting (85 K), supercooled (55 K), and quenched (20 K) liquid states, and in the crystal (20 K) state. Tagged pairs, initially lying in a given shell, were divided into incoming and outgoing groups and followed along simulated trajectories. Over them, specific correlation functions deltaB(r(0);t), involving the pair separation vector projected along its initial value r(0) (longitudinal dynamics), have been evaluated. More or less pronounced oscillations are detected according to the temperature of the thermodynamic states (and, obviously, their solid or liquid nature); for each state, they depend on the initial pair distance r(0), too. The oscillations vanish after few picoseconds (fast dynamics) in the case of crystal, whereas in the supercooled liquid they decay towards a plateau, whose height increases with the temperature. It is shown that the power spectrum of deltaB(r(0);t) practically yields the same density of states (DOS) produced by the pair velocity correlation function. The deltaB(r(0);t) functions obtained from the argon crystal at 20 K produce DOS curves dominated by two main frequency contributions, at about 40 and 60 cm(-1) (Einstein and Debye frequency, respectively). Their shape is quite well reproduced by damped harmonic oscillator like (DHO) functions vibrating at that frequencies. In liquid states, the deltaB(r(0);t) plateau, that forms after the fast DHO dynamics, accounts for the system diffusivity. The relaxation towards the plateau is modeled by an exponential function whose decay time is comparable with the average vibration period. Evidence that the liquid states conserve a certain memory of the vibrational modes of the crystal is obtained. In these states, the DHO functions at the Einstein and Debye frequencies plus an exponential function cannot reproduce the deltaB(r(0);t) shape. A pronounced shoulder, that forms around 0.5 ps, requires the contribution of a third DHO. In the DOS, it yields a band centered below 20 cm(-1) that produces low frequency DOS excess in comparison with the DOS of the crystal. This contribution is present in liquid and supercooled high temperature states and survives near the temperature of the glass transition whereas the diffusion practically vanishes. PMID- 14524757 TI - Simulation study on kinetic temperatures of vibrated binary granular mixtures. AB - We study the unequal kinetic temperatures in two-dimensional binary granular mixtures heated by vertical vibrations under gravity by molecular dynamics simulations. By introducing several variable parameters describing particle and composition properties of mixtures, the coexistence of two different kinetic temperatures for the two components is exhibited. In dilute situations, our numerical results confirm the overall experimental measurements by Feitosa and Menon recently [Phys. Rev. Lett. 88, 198301 (2002)]. For example, the temperature ratio along the vertical direction is constant in the bulk of a system, and is insensitive to external driving or relative concentration of the two components, but mainly influenced by their masses. However, outside of the dilute situations, especially as the number density becomes high enough where the spatial and momentum correlations become salient, the ratio profiles show some different characteristics. Generally, the plateau shape along the vertical direction is broken, and instead, a saddlelike shape appears. Furthermore, the physical implications are discussed. PMID- 14524758 TI - Slow dynamics and aging of a confined granular flow. AB - We present experimental results on slow flow properties of a granular assembly confined in a vertical column and driven upwards at a constant velocity V. The wall roughness is much lower than the typical grain size. For monodisperse assemblies this study evidences at low velocities (1>Gamma(s). The threshold Gamma(s) is shown to decrease when the bead is subjected, in simulations, to additional nondissipative collisions occurring with the typical frequency nu(c). As a consequence, the bead energy scales like v(2)(p)/(1-epsilon) for all vibration strengths in the limit nu(c)>>nu(*)(c). From the experimental and numerical findings, an analytical expression of the bead energy as a function of the experimental parameters is proposed. PMID- 14524762 TI - Statistical mechanics of granular gases in compartmentalized systems. AB - We study the behavior of an assembly of N granular particles contained in two compartments within a simple kinetic approach. The particles belonging to each compartment collide inelastically with each other and are driven by a stochastic heat bath. In addition, the fastest particles can change compartment at a rate that depends on their kinetic energy. Via a Boltzmann velocity distribution approach, we first study the dynamics of the model in terms of a coupled set of equations for the populations in the containers and their granular temperatures and find a crossover from a symmetric high-temperature phase to an asymmetric low temperature phase. Finally, in order to include statistical fluctuations, we solve the model within the direct simulation Monte Carlo approach. Comparisons with previous studies are presented. PMID- 14524764 TI - Initial susceptibility, flow curves, and magneto-optics of inverse magnetic fluids. AB - We introduce inverse magnetic fluids, consisting of gibbsite [Al(OH)(3)] platelets and alumina (Al2O3) spheres dispersed in a magnetic fluid, studied together with silica (SiO2) dispersions based on the same magnetic fluid matrix. Atomic force microscopy, optical microscopy, and alternate gradient magnetometry confirm the remarkable stability of the samples. Optical microscopy shows aggregation of nonmagnetic spheres, which, surprisingly, strongly depends on the concentration of the magnetic fluid rather than the concentration of nonmagnetic particles. Our model for the initial susceptibility of inverse magnetic fluids agrees very well with experimental data for systems containing spherical particles. The flow curves in an external magnetic field are strongly influenced by the aggregation of nonmagnetic particles or preformed nonmagnetic particle clusters, and by their disruption due to the shear flow. Static linear magnetobirefringence and magnetodichroism of all samples are investigated both experimentally and theoretically. These effects, which occur in all magnetic fluids, can be enhanced by the additional anisotropy due to the magnetic holes. The experiments we performed showed that, at a wavelength of 820 nm, the magnetodichroism is increased while the magneto-birefringence decreases when nonmagnetic particles were dispersed in the magnetic fluid. Magneto-birefringence is expected to be increased at large enough wavelengths only. PMID- 14524763 TI - Molecular arrangements of self-assembled surfactant films: characterization from atomic force microscopy data. AB - The adsorbed surfactant film molecular arrangement with thickness of approximately 5 nm is determined by measurements of the film dielectric permittivity. Before the advent of atomic force microscopy the dielectric permittivity was a macroscopic parameter, appropriate only for describing uniform environments since its profile was difficult to measure for local intermolecular interactions and its spatial distribution was frequently settled without experimental justification. Here, we show that atomic force microscopy made it possible to measure the dielectric permittivity profile in a scale below 5 nm for adsorbed layers of self-assembled surfactant films in water. The measured values of the film's dielectric permittivity and the film's thickness determine the compactness of the adsorbed film and consequently the presence of water molecules in the film and the conformal structure of the adsorbed molecules. PMID- 14524765 TI - Particle density stratification in transient sedimentation. AB - Theoretical predictions for the scaling of particle velocity fluctuations with container size in homogeneous Stokes suspensions are not consistent with experimental observations. Several explanations have been advanced, including the formation of stratification in bounded systems, such as those used in experiments. Numerical simulations of transient Stokes sedimentation in bounded cells are presented here for several cell sizes. The simulated cells have top and bottom wall boundaries and periodic boundaries in the horizontal. Throughout the course of the simulations the number and distribution of particles in the cell evolve, with impacts on the bulk mean particle velocity, velocity fluctuations, and particle density gradient. Initially the sedimentation follows the classical description, with a sharp front and uniform particle concentration below, but this is not sustained. A layer of higher particle concentration develops below the front. This is unstable and there is a large-scale overturning of the fluid. As a consequence, there is a redistribution of the particles, leaving behind a mass loading of the particles, which is stably stratified (subject to small density fluctuations between horizontal levels). The mean velocity and fluctuations of the particles initially grow and then decay once stable stratification has developed. PMID- 14524766 TI - Heterogeneity of colloidal particle networks analyzed by means of Minkowski functionals. AB - The heterogeneity and large scale connectivity of colloidal particle networks, which are generated by Brownian dynamics simulations, is examined. This is achieved by employing integral geometric measures in the form of the Minkowski functionals or quermassintegrals. It is found that these measures in conjunction with the parallel-body technique amount to a powerful tool to characterize the structure, going beyond the information contained in the pair-correlation function. The development of heterogeneities during network formation as well as their dependence on the volume fraction and the interaction potential is studied. In particular, it is found that slow coagulation enhances the heterogeneity of the network compared to fast coagulation. PMID- 14524767 TI - Logarithmic relaxation in a colloidal system. AB - The slow dynamics for a colloidal suspension of particles interacting with a hard core repulsion complemented by a short-ranged attraction is discussed within the frame of mode-coupling theory for ideal glass transitions for parameter points near a higher-order glass-transition singularity. The solutions of the equations of motion for the density correlation functions are solved for the square-well system in quantitative detail by asymptotic expansion using the distance of the three control parameters-packing fraction, attraction strength and attraction range-from their critical values as small parameters. For given wave vectors, distinguished surfaces in parameter space are identified where the next-to leading-order contributions for the expansion vanish so that the decay functions exhibit a logarithmic decay over large time intervals. For both coherent and tagged particle dynamics the leading-order logarithmic decay is accessible in the liquid regime for wave vectors of several times the principal peak in the structure factor. The logarithmic decay in the correlation function is manifested in the mean-squared displacement as a subdiffusive power law with an exponent varying sensitively with the control parameters. Shifting parameters through the distinguished surfaces, the correlation functions and the logarithm of the mean squared displacement considered as functions of the logarithm of the time exhibit a crossover from concave to convex behavior, and a similar scenario is obtained when varying the wave vector. PMID- 14524768 TI - Swelling-induced modulation of static and dynamic fluctuations in polyacrylamide gels observed by scanning microscopic light scattering. AB - The nature of inhomogeneities in vinylpolymer gels has hardly been clarified yet. Inhomogeneities on submicron and nanometer scales in polyacrylamide gels have been investigated by using a scanning microscopic light-scattering system and applying a general formula for an ensemble-averaged correlation function. The network structure of the gels is modified by varying the preparation conditions and can be roughly divided into two types. Swelling-induced modulation of inhomogeneities depends on the type of the network structure. At low monomer concentrations in preparation, both submicron- and nanometer-scale inhomogeneities increase with swelling. At high monomer concentrations in preparation, submicron-scale inhomogeneities increase with swelling, but nanometer-scale inhomogeneities decrease anomalously. This behavior is explained by a model of inhomogeneous network structure of vinylpolymer gels, where macrogel is formed from a large number of microgel particles. PMID- 14524769 TI - Photon correlation spectroscopy with high-energy coherent x rays. AB - We performed x-ray photon correlation spectroscopy on a model suspension of colloidal particles using x rays of three different energies, namely, 8 keV, 13.5 keV, and 19 keV. The observed reduction in the degree of coherence with increasing x-ray energy, as measured by the contrast of the correlation functions, is consistent with theoretical estimates. We show that it is well possible and under certain circumstances even advantageous to perform experiments with coherent x rays at these higher energies. We argue that the reduced absorption may not only allow for thicker samples but also for longer acquisition times because of the reduced radiation damage, thus outweighing in many cases the effect of the reduced coherent flux. The use of higher energy x rays for photon correlation spectroscopy can therefore lead to a substantial increase in the signal-to-noise ratio and constitutes a promising option for future experiments on samples of polymeric or biological origin. PMID- 14524770 TI - Long-range forces extending from polymer-gel surfaces. AB - Aqueous suspensions of microspheres were infused around gels of varying composition. The solutes were excluded from zones on the order of 100 microm from the gel surface. We present evidence that this finding is not an artifact, and that solute-repulsion forces exist at distances far greater than conventional theory predicts. The observations imply that solutes may interact over an unexpectedly long range. PMID- 14524771 TI - Difference and similarity of dielectric relaxation processes among polyols. AB - Complex permittivity measurements were performed on sorbitol, xylitol, and sorbitol-xylitol mixture in the supercooled liquid state in an extremely wide frequency range from 10 microHz to 500 MHz at temperatures near and above the glass transition temperature. We determined detailed behavior of the relaxation parameters such as relaxation frequency and broadening against temperature not only for the alpha process but also for the beta process above the glass transition temperature, to the best of our knowledge, for the first time. Since supercooled liquids are in the quasi-equilibrium state, the behavior of all the relaxation parameters for the beta process can be compared among the polyols as well as those for the alpha process. The relaxation frequencies of the alpha processes follow the Vogel-Fulcher-Tammann manner and the loci in the Arrhenius diagram are different corresponding to the difference of the glass transition temperatures. On the other hand, the relaxation frequencies of the beta processes, which are often called as the Johari-Goldstein processes, follow the Arrhenius-type temperature dependence. The relaxation parameters for the beta process are quite similar among the polyols at temperatures below the alphabeta merging temperature, T(M). However, they show anomalous behavior near T(M), which depends on the molecular size of materials. These results suggest that the origin of the beta process is essentially the same among the polyols. PMID- 14524772 TI - Entropy crisis, ideal glass transition, and polymer melting: exact solution on a Husimi cactus. AB - We investigate an extension of the lattice model of melting of semiflexible polymers originally proposed by Flory. Along with a bending penalty epsilon, present in the original model and involving three sites of the lattice, we introduce an interaction energy epsilon (p), corresponding to the presence of a pair of parallel bonds and an interaction energy epsilon (h), associated with a hairpin turn. Both these new terms represent four-site interactions. The model is solved exactly on a Husimi cactus, which approximates a square lattice. We study the phase diagram of the system as a function of the energies. For a proper choice of the interaction energies, the model exhibits a first-order melting transition between a liquid and a crystalline phase at a temperature T(M). The continuation of the liquid phase below T(M) gives rise to a supercooled liquid, which turns continuously into a new low-temperature phase, called metastable liquid, at T(MC)0 at fixed t reaches zero at a finite value of t. PMID- 14524775 TI - Wedge filling and interface delocalization in finite Ising lattices with antisymmetric surface fields. AB - Theoretical predictions by Parry et al. for wetting phenomena in a wedge geometry are tested by Monte Carlo simulations. Simple cubic LxLxL(y) Ising lattices with nearest neighbor ferromagnetic exchange and four free LxL(y) surfaces, at which antisymmetric surface fields +/-H(s) act, are studied for a wide range of linear dimensions (4 infinity and T larger than the filling transition temperature T(f)(H(s)), this interface runs from the one wedge where the surface planes with a different sign of the surface field meet (on average) straight to the opposite wedge, so that the average magnetization of the system is zero. For TT(f)(H(s)) from below, as is the corresponding behavior of the magnetization and its moments. We consider the variation of l(0) for T>T(f)(H(s)) as a function of a bulk field and find that the associated exponents agree with theoretical predictions. The correlation length xi(y) in the y direction along the wedges is also studied, and we find no transition for finite L and L(y)--> infinity. For L--> infinity the prediction l(0) proportional, variant (H(sc)-H(s))(-1/4) is verified, where H(sc)(T) is the inverse function of T(f)(H(s)) and xi(y) proportional, variant (H(sc)-H(s))( 3/4), respectively. We also find that m vanishes discontinuously at the filling transition. When the corresponding wetting transition is first order we also obtain a first-order filling transition. PMID- 14524776 TI - Hard-sphere fluids in contact with curved substrates. AB - The properties of a hard-sphere fluid in contact with hard-spherical and cylindrical walls are studied. Rosenfeld's density functional theory (DFT) is applied to determine the density profile and surface tension gamma for wide ranges of radii of the curved walls and densities of the hard-sphere fluid. Particular attention is paid to investigate the curvature dependence and the possible existence of a contribution to gamma which is proportional to the logarithm of the radius of curvature. Moreover, by treating the curved wall as a second component at infinite dilution, we provide an analytical expression for the surface tension of a hard-sphere fluid close to arbitrary hard convex walls. The agreement between the analytical expression and DFT is good. Our results show no signs for the existence of a logarithmic term in the curvature dependence of gamma. PMID- 14524777 TI - Orientational relaxation phenomena in Langmuir-Blodgett films at the air-water interface. AB - The nature of the orientational relaxation process of the director n to its equilibrium orientation n(eq), in the mono-(multi)layer(s) Langmuir-Blodgett film, during the lateral compression in absence of flow, is investigated. The relaxation time, during compression of 4-n-pentyl-4(')-cyanobiphenyl mono (multi)layer(s) film on the water surface, using the Ericksen-Leslie theory, has been calculated for the number of dynamic regimes. PMID- 14524778 TI - Relation between macroscopic and microscopic activation energies in nonequilibrium surface processing. AB - Realistic Monte Carlo simulations show that the apparent macroscopic activation energy is only partially explained by the expected expression for the average over the microscopic activation energies for surface processing. An additional term accounting for the existence of fluctuations in the fractions of particles has to be taken into account. In all cases considered, the additional term can be accurately estimated by a posteriori analysis of the temperature dependence of the surface densities. In addition, we demonstrate that the relative contribution of the different competing microscopic processes to the macroscopic activation energy can be accurately obtained during the simulations, allowing for the unambiguous identification of the particular surface species which effectively control the process. As an example of the nonequilibrium open interfaces to which the results apply, the case of wet chemical etching of crystalline silicon is considered. The results can be directly applied to surface growth. PMID- 14524779 TI - Surface freezing in binary mixtures of chain molecules. I. Alkane mixtures. AB - X-ray surface scattering and surface tension measurements are used to study surface freezing in molten mixtures of alkanes. These binary mixtures consist of protonated and deuterated alkanes, as well as of alkanes of different lengths. As for pure alkanes, a crystalline monolayer is formed at the surface a few degrees above the bulk freezing temperature. The structure of the monolayer has been determined on an angstrom scale. A simple theoretical approach is used to account for the thermodynamical observations at the surface and in the bulk. The model is based on a competition between entropic mixing and a repulsive interaction due to chain-length mismatch. The surface and bulk liquid phases are treated as ideal mixtures, while the solid phases are treated as regular mixtures. The theory is found to account well for all the mixtures studied, both hydrogenated hydrogenated and hydrogenated-deuterated. The repulsive interaction and its dependence on the chain lengths of the components are determined from fits to the measured data. PMID- 14524780 TI - Surface freezing in binary mixtures of chain molecules. II. Dry and hydrated alcohol mixtures. AB - Surface freezing is studied in dry and hydrated alcohol mixtures by surface x-ray scattering and surface tension measurements. A crystalline bilayer is formed at the surface a few degrees above the bulk freezing temperature. The packing is hexagonal, with molecules aligned along the surface normal in all cases. The in plane lattice constant reveals a qualitatively different behavior with composition for hydrated and dry mixtures. The simple theoretical approach used successfully for alkane and deuterated alkane mixtures accounts well also for the alcohol mixtures. The repulsive length-mismatch term opposing the mixing entropy term in the free energy of the mixtures is shown to have a universal behavior for all mixtures studied: protonated alkanes, deuterated alkanes, and dry and wet alcohols. This universality is somewhat counterintuitive in view of the different interactions (e.g., hydrogen bonding in alcohols) in the different mixtures. PMID- 14524781 TI - Growing surfaces with anomalous diffusion: results for the fractal Kardar-Parisi Zhang equation. AB - In this paper I study a model for a growing surface in the presence of anomalous diffusion, also known as the fractal Kardar-Parisi-Zhang equation (FKPZ). This equation includes a fractional Laplacian that accounts for the possibility that surface transport is caused by a hopping mechanism of a Levy flight. It is shown that for a specific choice of parameters of the FKPZ equation, the equation can be solved exactly in one dimension, so that all the critical exponents, which describe the surface that grows under FKPZ, can be derived for that case. Afterwards, the self-consistent expansion (SCE) is used to predict the critical exponents for the FKPZ model for any choice of the parameters and any spatial dimension. It is then verified that the results obtained using SCE recover the exact result in one dimension. At the end a simple picture for the behavior of the fractal KPZ equation is suggested and the upper critical dimension of this model is discussed. PMID- 14524782 TI - Surface-induced molecular tilt above the smectic-A-smectic-C phase transition in a nonchiral liquid crystal. AB - A polyimide-coated substrate was rubbed in such a way as to possess two competing easy axes for liquid crystal alignment. On cooling a homeotropically aligned liquid crystal through the smectic-A phase toward the smectic-C phase transition, an increasing tilt of the molecules relative to the layer normal was observed. The tilt was localized to within a smectic-C correlation length of the interface, and was found to increase monotonically with the rubbing strength associated with the preparation of the polyimide surface. The results are discussed in light of the dual easy axis model [T. Shioda et al., Phys. Rev. E 67, 041706 (2003)], and suggest that the two easy axes are not mutually orthogonal. PMID- 14524783 TI - Influence of flexoelectricity above the nematic Freedericksz transition. AB - Continuum theory is used to demonstrate that the presence of flexoelectricity significantly alters the response to an applied voltage of a homogeneous nematic liquid crystal cell above the ac Freedericksz threshold voltage. In such a system there is a fitting degeneracy: we obtain very good fits between theory and experimental permittivity data using any value of the sum of flexoelectric coefficients, e(11)+e(33), between 0.0 C/m and 1.5 x 10(-11) C/m. The corresponding values of the nematic bend elastic constant show an inverse parabolic relationship with e(11)+e(33), with K33 being reduced down to 90% of its value when flexoelectricity is neglected. PMID- 14524785 TI - Azimuthal and zenithal anchoring of nematic liquid crystals. AB - Temperature dependence of azimuthal and zenithal anchoring energy coefficients of the nematic liquid crystal 4-n-pentyl-4(')-cyanobiphenyl on rubbed nylon is measured using dynamic light scattering. The method is based on observations of director fluctuations in a planarly aligned wedge cell, where the anchoring energy coefficients can be obtained without any external torques acting on the liquid crystal during the measurement. We found that both anchoring coefficients decrease steadily on approaching the nematic-isotropic transition. Moreover, in the whole temperature range of the nematic phase, the ratio between the zenithal and the azimuthal anchoring coefficients is almost equal to the ratio between the splay and the twist Frank elastic constants. The same result is obtained also for the nematic phase of 4-n-octyl-4(')-cyanobiphenyl. This indicates that the aligning nylon layer directly affects only the monomolecular layer at the surface whereas the observed anchoring is governed by the elastic properties of the alkyl cyanobiphenyl. PMID- 14524784 TI - Light scattering investigation above the nematic-smectic-A phase transition in binary mixtures of calamitic and bent-core mesogens. AB - Quasielastic light scattering measurements were performed in the nematic phase of mixtures consisting of the calamitic mesogen 8OCB doped with small concentrations of the bent-core molecule P-7PIMB. It was found that the regular part of the bend elastic constant decreases strongly with dopant concentration X. Close to the nematic-smectic-A phase transition temperature, the divergent part of the bend elastic constant, which is proportional to the bare correlation length xi(0)(||) parallel to the layer normal, also decreases rapidly with X. The effect of the dopant on xi(0)(||) is examined in brief theoretically. PMID- 14524786 TI - Complex dielectric relaxation in supercooling and superpressing liquid crystalline chiral isopentylcyanobiphenyl. AB - Results of broadband dielectric studies in glass-forming liquid crystalline chiral isopentylcyanobiphenyl (5(*)CB) are presented. Tests conducted as a function of temperature and pressure revealed the coexistence of glassy and critical properties. The latter are associated with the isotropic-cholesteric phase transition at T(I-Ch) approximately 250 K under atmospheric pressure. Dielectric loss curves in the isotropic liquid and in the cholesteric phase are clearly broadened on cooling and pressuring towards the glass transition. Although in the isotropic phase there is a single stretched loss curve, in the mesophase an additional relaxation process can be distinguished. The evolution of relaxation times is non-Arrhenius and can be portrayed by the Vogel-Fulcher Tamman relation or its pressure counterpart. The glassy dynamics coexists with the critical-like behavior for the static dielectric permittivity and for the maxima of the dielectric loss curves. Their temperature and pressure dependences are associated with the critical exponent phi=1-alpha approximately 1/2, where alpha approximately 1/2 is the specific heat critical exponent. This behavior is associated with the continuous phase transition placed at DeltaT approximately 1.5 K below the clearing temperature for P=0.1 MPa. It has been found that 5(*)CB shows a unique pressure-temperature phase diagram. Pressure and temperature changes which begin in the isotropic liquid below at ca. T approximately 265 K always result in the transition to the cholesteric phase which can be supercooled or superpressed. For T>265 K the phase transition to another phase, presumably a solid one, always occurs. However, a cholesteric-solid phase border seems to exist only in isothermal pressure tests. It does not appear in the temperature studies. PMID- 14524787 TI - High-resolution x-ray study of the nematic-smectic-A and smectic-A-smectic-C transitions in liquid-crystal-aerosil gels. AB - The effects of dispersed aerosil nanoparticles on two of the phase transitions of the thermotropic liquid-crystal material 4-n-pentylphenylthiol-4(')-n octyloxybenzoate (8;S5) have been studied using high-resolution x-ray diffraction techniques. The aerosils hydrogen bond together to form a gel which imposes a weak quenched disorder on the liquid crystal. The smectic-A fluctuations are well characterized by a two-component line shape representing thermal and random-field contributions. An elaboration on this line shape is required to describe the fluctuations in the smectic-C phase; specifically the effect of the tilt on the wave-vector dependence of the thermal fluctuations must be explicitly taken into account. Both the magnitude and the temperature dependence of the smectic-C tilt order parameter are observed to be unaffected by the disorder. This may be a consequence of the large bare smectic correlation length in the direction of modulation for this transition. These results show that the understanding developed for the nematic to smectic-A transition for octylcyanobiphenyl and octyloxycyanobiphenyl liquid crystals with quenched disorder can be extended to quite different materials and transitions. PMID- 14524788 TI - Molecular orientation and the infrared dichroism of a chiral smectic liquid crystal in a homogeneously aligned cell at different temperature and bias fields. AB - The molecular orientation and the dichroic behavior of the vibrational bands of a homogeneously aligned helical cell containing chiral smectic liquid crystal (R)-( )-1-methylheptyl 4-(4(')-dodecyloxybiphenyl-4-ylcarbonyloxy)-3-fluorobenzoate are studied at various temperatures as a function of the bias field. These temperatures correspond to the various phase states of the sample at zero field. For those bands that exhibit significant dichroism, the field dependencies of the dichroic parameters (the dichroic ratio and the polarization angle of maximum absorbance) are found to be dependent on temperature, phase state, and helical unwinding. For the SmA* and SmC(*)(alpha) phases, the phenyl band dichroic ratio and the corresponding orientational order parameter are found to be almost independent of the bias field. The temperature dependence of the orientational order for zero field is discussed by taking into account the structures of the phases and the molecular tilt angles. The field dependencies of the phenyl band dichroic parameters for the SmC(*)(A) and SmC(*)(gamma) phases yield results about the distribution of directors in the layers of their unit cells and the state of helical unwinding. The azimuthal orientational distribution function of the carbonyl transition moments with respect to the long molecular axis has been determined. It is found that the degrees of the polar and quadrupolar biasing increase with decrease in temperature and the azimuthal biasing angle for the chiral carbonyl group increases significantly with a reduction in temperature. PMID- 14524789 TI - Dynamics of cholesteric structures in an electric field. AB - Motivated by Lehmann-like rotation phenomena in cholesteric drops we study the transverse drift of two types of cholesteric fingers, which form rotating spirals in thin layers of cholesteric liquid crystal in an ac or dc electric field. We show that electrohydrodynamic effects induced by Carr-Helfrich charge separation or flexoelectric charge generation can describe the drift of cholesteric fingers. We argue that the observed Lehmann-like phenomena can be understood on the same basis. PMID- 14524790 TI - Wetting properties of a hard-spherocylinder fluid on a substrate. AB - A density-functional theory is used to analyze the wetting properties of a fluid made up of hard spherocylinders of a length-to-breadth ratio L/D=5 on a model substrate. The substrate imposes an exclusion boundary condition over the molecular centres of mass, while at the same time favoring a definite molecular orientation, either parallel or perpendicular to the substrate, in a region next to the substrate. The wetting properties of this system are seen to depend on the strength with which the substrate orients the molecules: as the latter is increased, wetting by nematic phase is followed by a region of partial wetting which then leads to reentrant wetting by nematic. The two wetting transitions correspond to wetting films with nematic director perpendicular and parallel to the substrate, respectively. Also, in the region of partial wetting, an anchoring transition occurs in the substrate-nematic interface between two different director configurations (parallel and perpendicular to the substrate). Finally, a metastable wetting transition by isotropic is also obtained. This model considerably enriches the wetting phenomenology of the hard-spherocylinder fluid on substrates, of which only the pure hard wall, with no surface control parameter available, has been considered so far. PMID- 14524791 TI - Structure of the ripple phase of phospholipid multibilayers. AB - We present electron density maps (EDMs) of the ripple phase formed by phosphorylcholine lipids such as dimyristoyl phosphatidylcholine (DMPC), palmitoyl-oleoyl phosphatidylcholine (POPC), dihexadecyl phosphatidylcholine, and dilauroyl phosphatidylcholine (DLPC). With the exception of DLPC, the rippled bilayers have a sawtooth shape in all the systems, with one arm being almost twice as long as the other. For DMPC and POPC bilayers, EDMs have been obtained at different temperatures at a fixed relative humidity, and the overall shape of the ripples and the ratio of the lengths of the two arms are found to be insensitive to temperature. EDMs of all the systems with saturated hydrocarbon chains suggest the existence of a mean chain tilt along the ripple wave vector. In the literature it is generally assumed that the asymmetry of the rippled bilayers (absence of a mirror plane normal to the ripple wave vector) arises from a sawtoothlike height profile. However, in the case of DLPC, the height profile is found to be almost symmetric and the asymmetry results mainly from different bilayer thicknesses in the two arms of the ripple. We also present EDMs of the metastable ripple phase of dipalmitoyl phosphatidylcholine, formed on cooling from the L(alpha) phase. PMID- 14524792 TI - Phase diagram of poly(methyl-p-tolyl-siloxane): a temperature- and pressure dependent dielectric spectroscopy investigation. AB - The dynamics of poly(methyl-p-tolyl-siloxane) (PMpTS) have been studied as a function of temperature (in the range from 143 to 413 K), pressure (0.1-300 MPa), frequency (10(-2)-10(6) Hz), and molecular weight. Independent pressure-volume temperature (PVT) measurements (for temperatures in the range from 293 to 393 K and for pressures in the range from 10 to 200 MPa) allowed calculation of the relevant thermodynamic parameters. Two dielectrically active channels of relaxation were found, one in the glassy state reflecting a localized motion of the substituted phenyl ring and one at higher temperatures reflecting the usual segmental (alpha) relaxation. In PMpTS, there are two dominant control variables; both density and temperature have a strong influence on the segmental dynamics. The PVT results allowed us to follow distinct thermodynamic (T,P) paths resulting in states bearing the same density. These isodensity states are characterized by an apparent activation energy (Q(V)) that is not very different from the corresponding activation energy under isobaric conditions (Q(V)/Q(P) approximately 0.55) reflecting the importance of thermal effects. At temperatures above the glass temperature (T(g)), strong orientation correlations exist above some critical pressure that depends on temperature. This state extends from T(g) up to 1.08 T(g) and separates a normal liquid at higher temperatures from an oriented liquid at lower temperatures. Using the "phase diagram" we discuss separately the influence of the temperature and density on the PMpTS dynamics. PMID- 14524793 TI - Effect of volume changes on segmental relaxation in siloxane polymers. AB - From dielectric relaxation and equation-of-state measurements on polymethylphenylsiloxane and polymethyltolylsiloxane, the relative contributions of volume and thermal energy to the temperature dependence of the segmental relaxation times are quantified. In both polymers, volume exerts a substantial effect, being almost as significant as thermal energy. A possible consequence of this prominent role of volume in governing the segmental dynamics is the finding that the relaxation times, measured for a series of temperature at various (fixed) pressures, can be expressed as a single function of the volume normalized by the volume at the glass transition temperature. A similar result is found for the (isothermal) relaxation times measured at various pressures. PMID- 14524794 TI - Bottle-brush polymers as an intermediate between star and cylindrical polymers. AB - We present a theoretical study of a single bottle-brush molecule, which consists of multiarmed polymer stars grafted densely onto a stiff backbone. Mean-field approximation and a variational approach are used to calculate the dominant trajectories of the grafted chains, the shape of the molecule, and the segment density distribution around the backbone. All these properties are calculated for an arbitrary relationship between the size of the backbone and that of a grafted star. Hence cylindrical comb copolymer brushes and spherically symmetric polymer stars are considered as the limiting cases of the present problem. PMID- 14524795 TI - Polymer theta-point as a knot delocalization transition. AB - We study numerically the tightness of prime flat knots in a model of self attracting polymers with excluded volume. We find that these knots are localized in the high temperature swollen regime, but become delocalized in the low temperature globular phase. Precisely at the collapse transition, the knots are weakly localized. Some of our results can be interpreted in terms of the theory of polymer networks, which allows one to conjecture exact exponents for the knot length probability distributions. PMID- 14524796 TI - Out of equilibrium dynamics of poly(vinyl methyl ether) segments in miscible poly(styrene)-poly(vinyl methyl ether) blends. AB - The local dynamics of the low-T(g) component in a polymer blend, dynamically asymmetric poly(styrene)-poly(vinyl methyl ether) (PS-PVME), is studied below the glass transition, via dielectric relaxation spectroscopy. A particular attention has been paid to blends with a high PS content (PS weight fraction higher than 50%). A relaxation process, slower than the localized motions inducing the PVME secondary relaxations, is detected. Even though these blends fall out of equilibrium in this temperature regime, the structural recovery process is not efficient on the time scale of this PVME motional process. This relaxation is attributed to rather localized, weakly cooperative PVME motions resulting from the topological constraints imposed by the frozen PS chains. PMID- 14524797 TI - Direct observation of the effective bending moduli of a fluid membrane: free energy cost due to the reference-plane deformations. AB - Effective bending moduli of a fluid membrane are investigated by means of the transfer-matrix method developed in our preceding paper. This method allows us to survey various statistical measures for the partition sum. The role of the statistical measures is arousing much attention, since Pinnow and Helfrich claimed that under a suitable statistical measure, that is, the local mean curvature, the fluid membranes are stiffened, rather than softened, by thermal undulations. In this paper, we propose an efficient method to observe the effective bending moduli directly: We subjected a fluid membrane to a curved reference plane, and from the free-energy cost due to the reference-plane deformations, we read off the effective bending moduli. Accepting the mean curvature measure, we found that the effective bending rigidity gains even in the case of very flexible membrane (small bare rigidity); it has been rather controversial that for such a nonperturbative regime, the analytical prediction does apply. We also incorporate the Gaussian-curvature modulus and calculated its effective rigidity. Thereby, we found that the effective Gaussian-curvature modulus stays almost scale invariant. All these features are contrasted with the results under the normal-displacement measure. PMID- 14524798 TI - Competition between interchain and intrachain phase segregation. AB - Single-molecule observations of giant DNA have clarified that individual molecules undergo a marked discrete transition between an elongated coil state and a compact globule state. There is a relatively wide region of coexistence between the coil and the globule states, i.e., interchain phase segregation, with a change in intensive variables such as the concentration of the condensing agent, salt concentration, temperature. Very recently, the coexistence of coil and globule conformations within a single long DNA chain, i.e., intrachain phase segregation, has been reported under certain experimental conditions. In this study, we investigated general conditions for intrachain phase segregation in a single polyelectrolyte molecule, based on a simple statistical model. We consider the contribution of condensed counterions and the interaction energy of a charged coiled region. Intrachain phase segregation is stable with regard to free energy within a suitable parameter region. Our results suggest that intrachain phase segregation occurs when the electrostatic screening effect by the salt solution is negligible or when the screening effect is large and there is attractive interaction between polyelectrolyte segments. PMID- 14524799 TI - Dynamics of DNA in vitro evolution with Mnt-repressor: simulations and analysis. AB - The dynamics of DNA in vitro evolution with Mnt-repressor has been studied numerically and analytically. Based on experimental data and realistic energy landscape for DNA-Mnt-repressor interaction, we investigated the dynamics of DNA in vitro evolution using stochastic simulations. The binding energy of DNA to Mnt repressor was considered to consist of two parts: the DNA sequence specific and nonspecific. The crossover observed in real experiments is numerically recovered. We demonstrate that the evolution trajectories are drastically dispersed and no typical evolution passage exists during the evolution. Particularly, Fisher's theorem of natural selection is verified. A theoretical analysis for the evolution is also included. PMID- 14524800 TI - Increase in error threshold for quasispecies by heterogeneous replication accuracy. AB - In this paper we investigate the error threshold for quasispecies with heterogeneous replication accuracy. We show that the coexistence of error-free and error-prone polymerases can greatly increase the error threshold without a catastrophic loss of genetic information. We also show that the error threshold is influenced by the number of replicores. Our research suggests that quasispecies with heterogeneous replication accuracy can reduce the genetic cost of selective evolution while still producing a variety of mutants. PMID- 14524801 TI - Nuclear quantum effects on electron transfer reactions in DNA hairpins. AB - The driving force dependence of photoinduced electron transfer rate constant in synthetic DNA hairpins in aqueous solutions has been analyzed by means of molecular dynamics simulations. The quantum energy gap law has thus been investigated from a fully atomistic point of view, well reproducing the experimental results with reduced ambiguities in the parameter fitting. Although the contribution from the high-frequency vibrational modes of DNA and water solvent to the reorganization energy is fairly small, their quantum effect on the electron transfer rate constant is significant, well accounting for the deviation from the Marcus parabola observed in the experiments. PMID- 14524802 TI - Replica-free evaluation of the neuronal population information with mixed continuous and discrete stimuli: from the linear to the asymptotic regime. AB - Recent studies have explored theoretically the ability of populations of neurons to carry information about a set of stimuli, both in the case of purely discrete or purely continuous stimuli, and in the case of multidimensional continuous angular and discrete correlates, in the presence of additional quenched disorder in the distribution. An analytical expression for the mutual information has been obtained in the limit of large noise by means of the replica trick. Here, we show that the same results can actually be obtained in most cases without the use of replicas, by means of a much simpler expansion of the logarithm. Fitting the theoretical model to real neuronal data, we show that the introduction of correlations in the quenched disorder improves the fit, suggesting a possible role of signal correlations-actually detected in real data-in a redundant code. We show that even in the more difficult analysis of the asymptotic regime, an explicit expression for the mutual information can be obtained without resorting to the replica trick despite the presence of quenched disorder, both with a Gaussian and with a more realistic thresholded-Gaussian model. When the stimuli are mixed continuous and discrete, we find that with both models the information seem to grow logarithmically to infinity with the number of neurons and with the inverse of the noise, even though the exact general dependence cannot be derived explicitly for the thresholded-Gaussian model. In the large noise limit, lower values of information were obtained with the thresholded-Gaussian model, for a fixed value of the noise and of the population size. On the contrary, in the asymptotic regime, with very low values of the noise, a lower information value is obtained with the Gaussian model. PMID- 14524803 TI - Postinhibitory rebound delay and weak synchronization in Hodgkin-Huxley neuronal networks. AB - Noise-induced weak synchronized oscillatory activities in a globally inhibitory coupled Hodgkin-Huxley neuronal network are studied numerically. A kind of intrinsic delay induced by the postinhibitory rebound is observed and is found to be important in determining the overall frequency of the network. Synchronization occurs in an optimal range of noise intensity with a bell-shaped curve when the inhibitory coupling strength is sufficiently strong. Comparisons with the results for the excitatory coupling are also addressed. PMID- 14524804 TI - Anisotropic effects in highly scattering media. AB - In this paper, we study anisotropic scattering and light propagation models applicable to diffuse optical tomography. We propose a model for anisotropic scattering in the radiative transfer framework and derive the corresponding anisotropic diffusion model. To verify the anisotropic diffusion model, we consider the case of a simple anisotropic scattering model also presentable within the diffusion approximation. For numerical computations, we present a three-dimensional (3D) anisotropic Monte Carlo model and 2D finite element and boundary element solutions of the anisotropic diffusion model, and compare the results of the simulations. PMID- 14524805 TI - Recursiveness, switching, and fluctuations in a replicating catalytic network. AB - A protocell model consisting of mutually catalyzing molecules is studied, in order to investigate how chemical compositions are transferred recursively through cell divisions under replication errors. Depending on the numbers of molecules and species, and the path rate, three phases are found: fast switching state without recursive production, recursive production, and itinerancy between the above two states. The number distributions of the molecules in the recursive states are shown to be log-normal except for those species that form a core hypercycle, and are explained with the help of a heuristic argument. PMID- 14524806 TI - Synapse efficiency diverges due to synaptic pruning following overgrowth. AB - In the development of the brain, it is known that synapses are pruned following overgrowth. This pruning following overgrowth seems to be a universal phenomenon that occurs in almost all areas-visual cortex, motor area, association area, and so on. It has been shown numerically that the synapse efficiency is increased by systematic deletion. We discuss the synapse efficiency to evaluate the effect of pruning following overgrowth, and analytically show that the synapse efficiency diverges as O(|ln c|) at the limit where connecting rate c is extremely small. Under a fixed synapse number criterion, the optimal connecting rate, which maximizes memory performance, exists. PMID- 14524807 TI - Superparamagnetic segmentation by excitable neural systems. AB - Magnetic modeling for clustering or segmentation purposes can either associate the image data to external quenched fields or to the interactions among a set of auxiliary variables. The latter gives rise to superparamagnetic segmentation and is usually done with Potts systems. We have used the superparamagnetic clustering technique to segment images, with the aid of different associated systems. Results using Potts model are comparable to those obtained using excitable FitzHugh-Nagumo and Morris-Lecar model neurons. Interactions between the associated system components are a function of the difference of luminosity on a gray scale of neighbor pixels and the difference of membrane potential. PMID- 14524808 TI - Traveling waves with dispersive variability and time delay. AB - We first determine an approximate traveling wave profile for the Cook model [J. Murray, Mathematical Biology I: An Introduction (Springer, New York, 2002), pp. 471-478] for the case in which the number of dispersers is small relative to the number of nondispersers. The results are consistent with the previous linearized wavefront analysis that predicts, counterintuitively, that relatively few dispersers can drive the population expansion wave with a wavespeed not too different from that for the case of a single dispersing population as described by the Fisher equation. The method of solution differs from that used in the latter case since here the dimensionless wavespeed is close to unity. We next generalize the Cook model to include time-delay effects. While the Cook model, like the Fisher equation, does not adequately describe the wave of advance during the Neolithic transition in Europe, we show that the generalized Cook model provides a close agreement with the historical record. PMID- 14524809 TI - Punctuated equilibria and 1/f noise in a biological coevolution model with individual-based dynamics. AB - We present a study by linear stability analysis and large-scale Monte Carlo simulations of a simple model of biological coevolution. Selection is provided through a reproduction probability that contains quenched, random interspecies interactions, while genetic variation is provided through a low mutation rate. Both selection and mutation act on individual organisms. Consistent with some current theories of macroevolutionary dynamics, the model displays intermittent, statistically self-similar behavior with punctuated equilibria. The probability density for the lifetimes of ecological communities is well approximated by a power law with exponent near -2, and the corresponding power spectral densities show 1/f noise (flicker noise) over several decades. The long-lived communities (quasisteady states) consist of a relatively small number of mutualistically interacting species, and they are surrounded by a "protection zone" of closely related genotypes that have a very low probability of invading the resident community. The extent of the protection zone affects the stability of the community in a way analogous to the height of the free-energy barrier surrounding a metastable state in a physical system. Measures of biological diversity are on average stationary with no discernible trends, even over our very long simulation runs of approximately 3.4 x 10(7) generations. PMID- 14524810 TI - Wave nucleation rate in excitable systems in the low noise limit. AB - Motivated by recent experiments on intracellular calcium dynamics, we study the general issue of fluctuation-induced nucleation of waves in excitable media. We utilize a stochastic Fitzhugh-Nagumo model for this study, a spatially extended nonpotential pair of equations driven by thermal (i.e., white) noise. The nucleation rate is determined by finding the most probable escape path via minimization of an action related to the deviation of the fields from their deterministic trajectories. Our results pave the way both for studies of more realistic models of calcium dynamics as well as of nucleation phenomena in other nonequilibrium pattern-forming processes. PMID- 14524811 TI - Coexistence of stationary and traveling waves in reaction-diffusion-advection systems. AB - The flow- and diffusion-distributed structures (FDS) and the differential-flow instability (DIFI) are mechanisms that give rise to static and traveling waves in reactive flows with general, species-dependent transport terms. Here we consider a general framework which supports the simultaneous existence of FDS and DIFI patterns. We study the necessary conditions for each instability in general and compare them in order to derive their connection. The interaction between FDS and DIFI patterns gives rise to interesting wave behavior including stationary, upstream, and downstream traveling waves as well as an interesting regime where stationary and traveling waves coexist. PMID- 14524812 TI - Non-Gaussian probability distribution functions from maximum-entropy-principle considerations. AB - In this work we develop the recently suggested concept of superstatistics [C. Beck and E.G.D. Cohen, Physica A 322, 267 (2003)], face the problem of devising a viable way for estimating the correct statistics for a system in the absence of sufficient knowledge of its microscopical dynamics, and suggest to solve it through the maximum-entropy principle. As an example, we deduce the probability distribution function for velocity fluctuations in turbulent fluids, which is slightly different from the form suggested by C. Beck [Phys. Rev. Lett. 87, 180601 (2001)]. PMID- 14524813 TI - Test of nonequilibrium thermodynamics in glassy systems: the soft-sphere case. AB - The scaling properties of the soft-sphere potential allow the derivation of an exact expression for the pressure of a frozen liquid, i.e., the pressure corresponding to configurations which are local minima in its multidimensional potential energy landscape. The existence of such a relation offers the unique possibility for testing the recently proposed extension of the liquid free energy to glassy out-of-equilibrium conditions and the associated expression for the temperature of the configurational degrees of freedom. We demonstrate that the nonequilibrium free energy provides an exact description of the soft-sphere pressure in glass states. PMID- 14524814 TI - Partial rejuvenation of a colloidal glass. AB - We study the effect of shear on the aging dynamics of a colloidal suspension of synthetic clay particles. We find that a shear of amplitude gamma reduces the relaxation time measured just after the cessation of shear by a factor exp( gamma/gamma(c)), with gamma(c) approximately 5%, and is independent of the duration and the frequency of the shear. This simple law for the rejuvenation effect shows that the energy involved in colloidal rearrangements is proportional to the shear amplitude gamma rather than gamma(2), leading to an Eyring-like description of the dynamics of our system. PMID- 14524815 TI - Wave number selection in a nonequilibrium electro-osmotic instability. AB - Nonequilibrium electro-osmotic slip causes instability of quiescent ionic conductance through a diffusion layer of a strong electrolyte at a charge selective solid such as ion-exchange membrane or electrode. This instability, as inferred from the outer asymptotic limit of the full singularly perturbed ionic transport problem, is of the short-wave type. This latter is a serious shortcoming of the limiting model. In this Brief Report we show that inclusion in the model of the first asymptotic corrections yields a reasonable finite wavelength selection. PMID- 14524816 TI - Comment on "Towards deterministic equations for Levy walks: the fractional material derivative". AB - The connection of problems considered in the paper by Sokolov and Metzler with stochastic transport in usual space and uniform medium is discussed. PMID- 14524817 TI - Phase-separation transition in one-dimensional driven models. AB - A class of models of two-species driven diffusive systems which is shown to exhibit phase separation in d=1 dimensions is introduced. Unlike previously studied models exhibiting similar phenomena, here the relative density of the two species is fluctuating within the macroscopic domain of the phase separtated state. The nature of the phase transition from the homogeneous to the phase separated state is discussed in view of a recently introduced criterion for phase separation in one-dimensional driven systems. PMID- 14524818 TI - Local distributions and rate fluctuations in a unified scaling law for earthquakes. AB - A recently proposed unified scaling law for interoccurrence times of earthquakes is analyzed, both theoretically and with data from Southern California. We decompose the corresponding probability density into local-instantaneous distributions, which scale with the rate of earthquake occurrence. The fluctuations of the rate, characterizing the nonstationarity of the process, show a double power-law distribution and are fundamental to determine the overall behavior, described by a double power law as well. PMID- 14524819 TI - Epidemic incidence in correlated complex networks. AB - We introduce a numerical method to solve epidemic models on the underlying topology of complex networks. The approach exploits the mean-field-like rate equations describing the system and allows us to work with very large system sizes, where Monte Carlo simulations are useless due to memory needs. We then study the susceptible-infected-removed epidemiological model on assortative networks, providing numerical evidence of the absence of epidemic thresholds. Besides, the time profiles of the populations are analyzed. Finally, we stress that the present method would allow us to solve arbitrary epidemiclike models provided that they can be described by mean-field rate equations. PMID- 14524820 TI - Thermodynamic relations in a driven lattice gas: numerical experiments. AB - We explore thermodynamic relations in nonequilibrium steady states with numerical experiments on a driven lattice gas. After operationally defining the pressure and chemical potential in the driven lattice gas, we confirm numerically the validity of the integrability condition (the Maxwell relation) for the two quantities whose values differ from those for an equilibrium system. This implies that a free-energy function can be constructed for the nonequilibrium steady state that we consider. We also investigate a fluctuation relation associated with this free-energy function. Our result suggests that the compressibility can be expressed in terms of density fluctuations even in nonequilibrium steady states. PMID- 14524821 TI - Orientational preference and predictability in a symmetric arrangement of magnetic drops. AB - An investigation of a symmetrical arrangement of N quasi-two-dimensional magnetic domains in an external field is carried out. By minimizing the linearized interaction energy for this arrangement using a nearest-neighbor approximation, an orientationally preferred state of the system is found. This orientational preference leads to a large degree of predictability in the final patterns as demonstrated by some experiments using ferrofluids. The final state patterns are also investigated by carrying out a series of numerical simulations. These simulations exhibit a similar predictability and the final patterns bear a strong resemblance to those obtained experimentally. PMID- 14524822 TI - Discrete velocity and lattice Boltzmann models for binary mixtures of nonideal fluids. AB - In this paper, a discrete velocity model and a lattice Boltzmann model are proposed for binary mixtures of nonideal fluids based on the Enskog theory. The velocity space of the Enskog equation for each component is first discretized by applying a Gaussian quadrature, resulting in a discrete velocity model that can be solved by suitable numerical schemes. A lattice Boltzmann model is then derived from the discrete velocity model with a slightly modified equilibrium. The hydrodynamics of each model are also derived through the Chapmann-Enskog procedure. PMID- 14524823 TI - Direct observation of period-doubled nonspherical states in single-bubble sonoluminescence. AB - We present direct observations of period doubling in the flash to flash pulse heights in single-bubble sonoluminescence. States involved are stable, spherically symmetry broken. Observations are made using seven detectors distributed in the equatorial plane of the bubble. Contrary to earlier experiments by Holt et al. [Phys. Rev. Lett. 72, 1376 (1994)], where period doubling was observed in the time intervals between flashes but not in the pulse heights, we observe period doubling in pulse heights, but no corresponding period doubling is seen in the time intervals. In parameter space the period doubling is observed below the n=2 shape instability boundary line where extinction is shown to take place. PMID- 14524824 TI - Polarization of wave modes in a two-dimensional hexagonal lattice using a complex (dusty) plasma. AB - Wave spectra corresponding to the random particle motion in a monolayer Yukawa crystal were studied for various directions of propagation. It was found that there are two wave modes with a polarization alternating between the longitudinal and transverse. In the long-wavelength regime, the modes became purely longitudinal and transverse as was known before. In the short-wavelength regime the spectra strongly depended on the wavelength and the direction of propagation. The results obtained from the experiment, theory, and simulation agreed well with each other. PMID- 14524825 TI - Plasma channels produced by a laser-triggered high-voltage discharge. AB - A plasma waveguide scheme for high-intensity laser guiding with densities and lengths suitable for laser-plasma particle accelerators is presented. This scheme uses a laser-triggered high-voltage discharge, presents negligible jitter, allows full access to the plasma, and can be scaled to large distances. Experimental results showing the feasibility of this scheme are presented. PMID- 14524826 TI - Onset of incommensurate interfacial instability in a minimal model of dry friction. AB - We present a minimal model of dry friction between two incommensurate interfaces sliding at high relative velocity. Many of the features of the friction force for the full two-dimensional many-body dynamical system-particularly in the sub critical velocity regime-are captured by our one-dimensional Einstein model, where the motion of a typical interfacial atom is constrained to be vertical to the sliding plane. Beyond the linear response of force versus sliding velocity, the anharmonic Einstein model predicts a doublet resonance peak, whereupon a catastrophe in the model signals the onset of a plastic deformation mechanism for frictional sliding, namely, the instability of the interface. Higher velocities than this critical value require a much more sophisticated description of the production and coalescence of dislocations into a microstructure. PMID- 14524827 TI - Exact soliton solution and inelastic two-soliton collision in a spin chain driven by a time-dependent magnetic field. AB - We investigate dynamics of exact N-soliton trains in a spin chain driven by a time-dependent magnetic field by means of an inverse scattering transformation. The one-soliton solution indicates obviously the spin precession around the magnetic field and periodic shape variation induced by the time-varying field as well. In terms of the general soliton solutions, N-soliton interaction and particularly various two-soliton collisions are analyzed. The inelastic collision by which we mean the soliton shape change before and after collision appears is generally due to the time-varying field. We, moreover, show that complete inelastic collisions can be achieved by adjusting spectrum and field parameters. This may lead to a potential technique of shape control of soliton. PMID- 14524828 TI - Epidemic processes with immunization. AB - We study a model of directed percolation (DP) with immunization, i.e., with different probabilities for the first infection and subsequent infections. The immunization effect leads to an additional non-Markovian term in the corresponding field theoretical action. We consider immunization as a small perturbation around the DP fixed point in d<6, where the non-Markovian term is relevant. The immunization causes the system to be driven away from the neighborhood of the DP critical point. In order to investigate the dynamical critical behavior of the model, we consider the limits of low and high first infection rate, while the second-infection rate remains constant at the DP critical value. Scaling arguments are applied to obtain an expression for the survival probability in both limits. The corresponding exponents are written in terms of the critical exponents for ordinary DP and DP with a wall. We find that the survival probability does not obey a power-law behavior, decaying instead as a stretched exponential in the low first-infection probability limit and to a constant in the high first-infection probability limit. The theoretical predictions are confirmed by optimized numerical simulations in 1+1 dimensions. PMID- 14524829 TI - Thermodynamics of critical strange nonchaotic attractors. AB - The thermodynamic formalism is applied to dynamical attractors which have fractal geometry and on which all Lyapunov exponents are exactly zero. Such critical strange nonchaotic attractors (SNAs) which arise, for example, in the Harper system exhibit a static phase transition in the free energy. The Tsallis nonextensive entropy which is known to characterize the thermodynamics of systems with leading Lyapunov exponent zero is found to be subadditive for the critical states. These properties are shared by other quasiperiodic systems with critical SNAs. PMID- 14524830 TI - Large-scale structural organization of social networks. AB - The characterization of large-scale structural organization of social networks is an important interdisciplinary problem. We show, by using scaling analysis and numerical computation, that the following factors are relevant for models of social networks: the correlation between friendship ties among people and the position of their social groups, as well as the correlation between the positions of different social groups to which a person belongs. PMID- 14524831 TI - Searching in small-world networks. AB - We study the average time it takes to find a desired node in the Watts-Strogatz family of networks. We consider the case when the look-up time can be neglected and when it is important, where the look-up time is the time needed to choose one among all the neighboring nodes of a node at each step in the search. We show that in both cases, the search time is minimum in the small-world regime, when an appropriate distance between the nodes is defined. Through an analytical model, we show that the search time scales as N(1/D(D+1)) for small-world networks, where N is the number of nodes and D is the dimension of the underlying lattice. This model is shown to be in agreement with numerical simulations. PMID- 14524832 TI - Fluctuation of riding passengers induced by chaotic motions of shuttle buses. AB - We study the fluctuation of the number of riding passengers in a few shuttle buses that pass each other freely. We present a dynamical model of the shuttle buses that takes into account the maximum capacity of a bus. The dynamics of the buses is expressed in terms of a coupled nonlinear map with noise. The number of passengers carried by a bus and the time headway between buses exhibit complex behavior with varying trips. It is found that the behavior of the buses exhibits deterministic chaos even if there is no noise. The chaotic motion depends on the loading parameter, the maximum capacity of a bus, the bus's speed, and the number of buses. When the loading parameter is larger than a threshold value, each bus carries a full load of passengers throughout its trip. The dependence of the threshold (transition point) on both capacity and speed is clarified. PMID- 14524833 TI - Generalized thermodynamics and Fokker-Planck equations: applications to stellar dynamics and two-dimensional turbulence. AB - We introduce a class of generalized Fokker-Planck equations that conserve energy and mass and increase a generalized entropy functional until a maximum entropy state is reached. Nonlinear Fokker-Planck equations associated with Tsallis entropies are a special case of these equations. Applications of these results to stellar dynamics and vortex dynamics are proposed. Our prime result is a relaxation equation that should offer an easily implementable parametrization of two-dimensional turbulence. Usual parametrizations (including a single turbulent viscosity) correspond to the infinite temperature limit of our model. They forget a fundamental systematic drift that acts against diffusion as in Brownian theory. Our generalized Fokker-Planck equations can have applications in other fields of physics such as chemotaxis for bacterial populations. We propose the idea of a classification of generalized entropies in "classes of equivalence" and provide an aesthetic connection between topics (vortices, stars, bacteria, em leader ) which were previously disconnected. PMID- 14524834 TI - Investigation of the forest-fire model on a small-world network. AB - It is shown that the forest-fire model of Bak et al. run on a square lattice network with additional long-range interactions in the spirit of a small-world network results in a scale-free system reminiscent of self-organized criticality without recourse to fine tuning. As the number of these long-range interactions is increased, the cluster size distribution exponent is found to decrease in magnitude as the small-world regime is entered, indicating a change in its universality class. It is suggested that such a model could have applicability in the study of disease spreading in human populations. PMID- 14524835 TI - Solid-liquid transition of ultrathin lubricant film. AB - We represent a melting of ultrathin lubricant film by friction between atomically flat surfaces as a result of action of spontaneously appearing elastic field of stress shear component caused by the external supercritical heating. The kinetics of this solid-liquid transition is described by the Maxwell-type and Voigt-Kelvin equations for viscoelastic matter as well as by the relaxation equation for temperature. We show that these equations coincide formally with the synergetic Lorenz system, where the stress acts as the order parameter, the conjugate field is reduced to the elastic shear strain, and the temperature is the control parameter. Using the adiabatic approximation we find the steady-state values of these quantities. Taking into account the deformational defect of the shear modulus, we show that lubricant melting is realized according to mechanism of the first-order transition. The critical temperature of the friction surfaces increases with growth of the characteristic value of shear viscosity and decreases with growth of the shear modulus value linearly. PMID- 14524836 TI - Critical behavior and scaling functions of the three-dimensional O(6) model. AB - We numerically investigate the three-dimensional O(6) model on 12(3) to 120(3) lattices within the critical region at zero magnetic field, as well as at finite magnetic field on the critical isotherm and for several fixed couplings in the broken and the symmetric phase. We obtain from the Binder cumulant at vanishing magnetic field the critical coupling J(c)=1.42865(3). The universal value of the Binder cumulant at this point is g(r)(J(c))=-1.94456(10). At the critical coupling, the critical exponents gamma=1.604(6), beta=0.425(2), and nu=0.818(5) are determined from a finite-size-scaling analysis. Furthermore, we verify predicted effects induced by massless Goldstone modes in the broken phase. The results are well described by the perturbative form of the model's equation of state. Our O(6) result is compared to the corresponding Ising, O(2) and O(4) scaling functions. Finally, we study the finite-size-scaling behavior of the magnetization on the pseudocritical line. PMID- 14524837 TI - Class of correlated random networks with hidden variables. AB - We study a class of models of correlated random networks in which vertices are characterized by hidden variables controlling the establishment of edges between pairs of vertices. We find analytical expressions for the main topological properties of these models as a function of the distribution of hidden variables and the probability of connecting vertices. The expressions obtained are checked by means of numerical simulations in a particular example. The general model is extended to describe a practical algorithm to generate random networks with an a priori specified correlation structure. We also present an extension of the class, to map nonequilibrium growing networks to networks with hidden variables that represent the time at which each vertex was introduced in the system. PMID- 14524838 TI - Universality in the pair contact process with diffusion. AB - The pair contact process with diffusion is studied by means of multispin Monte Carlo simulations and density matrix renormalization group calculations. Effective critical exponents are found to behave nonmonotonically as functions of time or of system length and extrapolate asymptotically towards values consistent with the directed percolation universality class. We argue that an intermediate regime exists where the effective critical dynamics resembles that of a parity conserving process. PMID- 14524839 TI - Anomalous diffusion, localization, aging, and subaging effects in trap models at very low temperature. AB - We study in detail the dynamics of the one-dimensional symmetric trap model via a real-space renormalization procedure which becomes exact in the limit of zero temperature. In this limit, the diffusion front in each sample consists of two delta peaks, which are completely out of equilibrium with each other. The statistics of the positions and weights of these delta peaks over the samples allows to obtain explicit results for all observables in the limit T-->0. We first compute disorder averages of one-time observables, such as the diffusion front, the thermal width, the localization parameters, the two-particle correlation function, and the generating function of thermal cumulants of the position. We then study aging and subaging effects: our approach reproduces very simply the two different aging exponents and yields explicit forms for scaling functions of the various two-time correlations. We also extend the real-space renormalization group method to include systematic corrections to the previous zero temperature procedure via a series expansion in T. We then consider the generalized trap model with parameter alpha in [0,1] and obtain that the large scale effective model at low temperature does not depend on alpha in any dimension, so that the only observables sensitive to alpha are those that measure the "local persistence," such as the probability to remain exactly in the same trap during a time interval. Finally, we extend our approach at a scaling level for the trap model in d=2 and obtain the two relevant time scales for aging properties. PMID- 14524840 TI - Classical infinite-range-interaction Heisenberg ferromagnetic model: metastability and sensitivity to initial conditions. AB - An N-sized inertial classical Heisenberg ferromagnet, which consists of a modification of the well-known standard model, where the spins are replaced by classical rotators, is studied in the limit of infinite-range interactions. The usual canonical-ensemble mean-field solution of the inertial classical n-vector ferromagnet (for which n=3 recovers the particular Heisenberg model considered herein) is briefly reviewed, showing the well-known second-order phase transition. This Heisenberg model is studied numerically within the microcanonical ensemble through molecular dynamics. In what concerns the caloric curve, it is shown that, far from criticality, the kinetic temperature obtained at the long-time-limit microcanonical-ensemble simulation recovers well the equilibrium canonical-ensemble estimate, whereas, close to criticality, a discrepancy (presumably due to finite-size effects) is found. The time evolution of the kinetic temperature indicates that a basin of attraction exists for the initial conditions for which the system evolves into a metastable state, whose duration diverges as N--> infinity, before attaining the terminal thermal equilibrium. Such a metastable state is observed for a whole range of energies, which starts right below criticality and extends up to very high energies (in fact, the gap between the kinetic temperatures associated with the metastable and the terminal-equilibrium states is expected to disappear only as one approaches infinite energy). To the best our knowledge, this has never before been observed on similar Hamiltonian models, in a noticeable way, for such a large range of energies. For example, for the XY (n=2) version of the present model, such a behavior was observed only near criticality. It is shown also that the (metastable state) maximum Lyapunov exponent decreases with N like lambda(max) approximately N-kappa, where for the initial conditions employed herein (maximal magnetization), kappa=0.225+/-0.030, both above and below the critical point. PMID- 14524841 TI - Phase properties of a zigzag chain lattice gas with Coulomb interactions. AB - It has been reported that the phase transitions found in the quasi-one dimensional sulfide KCu(7-x)S4 are most likely due to vacancy ordering involving Cu+-ion diffusion along the Cu(2)-Cu(2) zigzag chains. Our previous studies with both a self-consistent method and Monte Carlo simulations confirmed that phase transitions indeed exist in a one-dimensional (1D) lattice gas system in which vacancy ordering is involved. In this paper, we calculate the more nearly real case of KCu6.88S4 and further investigate the angular dependence of the phase properties in a partially occupied 1D zigzag chain with various particle occupancies. The calculated results suggest that the phase transitions that occur in the quasi-one-dimensional material KCu(7-x)S4 are presumably due to both intrachain and interchain interactions between the partially occupied Cu+ zigzag chains. Most interestingly, we found that the average particle distribution of the lowest free energy state is a linear superposition of two other solutions with different particle distributions for occupancy n(av)=1/2. PMID- 14524842 TI - Collapses and explosions in self-gravitating systems. AB - Collapse and explosion (reverse to collapse) transitions in self-gravitating systems are studied by molecular dynamics simulations. A microcanonical ensemble of point particles confined to a spherical box is considered. The particles interact via an attractive soft Coulomb potential. It is observed that a collapse indeed takes place when the energy of the uniform state is set near or below the metastability-instability threshold (collapse energy) as predicted by the mean field theory. Similarly, an explosion occurs when the energy of the core-halo state is increased above the explosion energy, where according to the mean-field predictions the core-halo state becomes unstable. For systems consisting of 125 500 particles, the collapse takes about 10(5) single-particle crossing times to complete, while a typical explosion is by an order of magnitude faster. A finite lifetime of metastable states is observed. It is also found that the mean-field description of the uniform and core-halo states is exact within the statistical uncertainty of the molecular dynamics data. PMID- 14524843 TI - Does the continuum theory of dynamic fracture work? AB - We investigate the validity of the linear elastic fracture mechanics approach to dynamic fracture. We first test the predictions in a lattice simulation, using a formula of Eshelby for the time-dependent stress intensity factor. Excellent agreement with the theory is found. We then use the same method to analyze the experiment of Sharon and Fineberg. The data here are not consistent with the theoretical expectation. PMID- 14524844 TI - Correlations in scale-free networks: tomography and percolation. AB - We discuss three related models of scale-free networks with the same degree distribution but different correlation properties. Starting from the Barabasi Albert construction based on growth and preferential attachment we discuss two other networks emerging when randomizing it with respect to links or nodes. We point out that the Barabasi-Albert model displays dissortative behavior with respect to the nodes' degrees, while the node-randomized network shows assortative mixing. These kinds of correlations are visualized by discussing the shell structure of the networks around an arbitrary node. In spite of different correlation behaviors, all three constructions exhibit similar percolation properties. This result for percolation is also detected for a network with finite second moment and its corresponding randomized models. PMID- 14524845 TI - Lyapunov exponent of many-particle systems: testing the stochastic approach. AB - The stochastic approach to the determination of the largest Lyapunov exponent of a many-particle system is tested in the so-called mean-field XY Hamiltonians. In weakly chaotic regimes, the stochastic approach relates the Lyapunov exponent to a few statistical properties of the Hessian matrix of the interaction, which can be calculated as suitable thermal averages. We have verified that there is a satisfactory quantitative agreement between theory and simulations in the disordered phases of the XY models, either with attractive or repulsive interactions. Part of the success of the theory is due to the possibility of predicting the shape of the required correlation functions, because this permits the calculation of correlation times as thermal averages. PMID- 14524846 TI - Surface growth models with a random-walk-like nonlocality. AB - To understand the effects of a random-walk-like nonlocality on the dynamical scaling properties of surface growths, a stochastic growth model in which the height difference triangle up h((i,i+1))=|h(i)-h(i+1)| of a chosen nearest neighbor column pair (i,i+1) is decreased by one unit is introduced and studied by simulations. The probability P((i,i+1)) of choosing a column pair (i,i+1) on a one-dimensional substrate is assigned as P((i,i+1))=e(kappa triangle up h((i,i+1)))/ summation operator (L)(j=1)e(kappa triangle up h((j,j+1))). On a substrate of given size L, the dynamical scaling property satisfies a normal scaling behavior as W=L(alpha)f(t/L(z)), when kappa is very small. If kappa becomes moderately large, the scaling property with the dynamic exponent z=1 as in diffusion-limited erosion appears. If kappa becomes very large, no surface roughening is found. PMID- 14524847 TI - Why social networks are different from other types of networks. AB - We argue that social networks differ from most other types of networks, including technological and biological networks, in two important ways. First, they have nontrivial clustering or network transitivity and second, they show positive correlations, also called assortative mixing, between the degrees of adjacent vertices. Social networks are often divided into groups or communities, and it has recently been suggested that this division could account for the observed clustering. We demonstrate that group structure in networks can also account for degree correlations. We show using a simple model that we should expect assortative mixing in such networks whenever there is variation in the sizes of the groups and that the predicted level of assortative mixing compares well with that observed in real-world networks. PMID- 14524848 TI - Observing stochastic resonance in an underdamped bistable Duffing oscillator by the method of moments. AB - The method of moments is applied to an underdamped bistable oscillator driven by Gaussian white noise and a weak periodic force for the observations of stochastic resonance and the resulting resonant structures are compared with those from Langevin simulation. The physical mechanisms of the stochastic resonance are explained based on the evolution of the intrawell frequency peak and the above barrier frequency peak via the noise intensity and the fluctuation-dissipation theorem, and the three possible sources of stochastic resonance in the system are confirmed. Additionally, with the noise intensity fixed, the stochastic resonant structures are also observed by adjusting the nonlinear parameter. PMID- 14524849 TI - Urban economy as a scale-free network. AB - We present empirical evidence that land values are scale free and introduce a network model that reproduces the observations. The network approach to urban modeling is based on the assumption that the market dynamics that generates land values can be represented as a growing scale-free network. Our results suggest that the network properties of trade between specialized activities cause land values, and likely also other observables such as population, to be power-law distributed. In addition to being an attractive avenue for further analytical inquiry, the network representation is also applicable to empirical data and is thereby attractive for predictive modeling. PMID- 14524850 TI - Simultaneous analysis of several models in the three-dimensional Ising universality class. AB - We investigate several three-dimensional lattice models believed to be in the Ising universality class by means of Monte Carlo methods and finite-size scaling. These models include spin-1 / 2 models with nearest-neighbor interactions on the simple-cubic and on the diamond lattice. For the simple cubic lattice, we also include models with third-neighbor interactions of varying strength, and some "equivalent-neighbor" models. Also included are a spin-1 model and a hard-core lattice gas. Separate analyses of the numerical data confirm the Ising-like critical behavior of these systems. On this basis, we analyze all these data simultaneously such that the universal parameters occur only once. This leads to an improved accuracy. The thermal, magnetic, and irrelevant exponents are determined as y(t)=1.5868(3), y(h)=2.4816(1), and y(i)=-0.821(5), respectively. The Binder ratio is estimated as Q=(2)/=0.62 341(4). PMID- 14524851 TI - Multioverlap simulations for transitions between reference configurations. AB - We introduce a procedure to construct weight factors, which flatten the probability density of the overlap with respect to some predefined reference configuration. This allows one to overcome free-energy barriers in the overlap variable. Subsequently, we generalize the approach to deal with the overlaps with respect to two reference configurations so that transitions between them are induced. We illustrate our approach by simulations of the brain peptide Met enkephalin with the ECEPP/2 (Empirical Conformational Energy Program for Peptides) energy function using the global-energy-minimum and the second lowest energy states as reference configurations. The free energy is obtained as functions of the dihedral and the root-mean-square distances from these two configurations. The latter allows one to identify the transition state and to estimate its associated free-energy barrier. PMID- 14524852 TI - Car accidents in cellular automata models for one-lane traffic flow. AB - Conditions for the occurrence of car accidents are introduced in the Nagel Schreckenberg model. These conditions are based on the thought that a real accident depends on several parameters: an unexpected action of the car ahead (sudden stop or abrupt deceleration), the gap between the two cars, the velocity of the successor car and its delayed reaction time. We discuss then the effect of this delayed reaction time on the probability of traffic accidents. We find that these conditions for the occurrence of car accidents are necessary for modeling realistic accidents. PMID- 14524853 TI - Universal interface width distributions at the depinning threshold. AB - We compute the probability distribution of the interface width at the depinning threshold, using recent powerful algorithms. It confirms the universality classes found previously. In all cases, the distribution is surprisingly well approximated by a generalized Gaussian theory of independent modes which decay with a characteristic propagator G(q)=1/q(d+2zeta); zeta, the roughness exponent, is computed independently. A functional renormalization analysis explains this result and allows one to compute the small deviations, i.e., a universal kurtosis ratio, in agreement with numerics. We stress the importance of the Gaussian theory to interpret numerical data and experiments. PMID- 14524854 TI - Logarithmic corrections to scaling in critical percolation and random resistor networks. AB - We study the critical behavior of various geometrical and transport properties of percolation in six dimensions. By employing field theory and renormalization group methods we analyze fluctuation induced logarithmic corrections to scaling up to and including the next-to-leading order correction. Our study comprehends the percolation correlation function, i.e., the probability that two given points are connected, and some of the fractal masses describing percolation clusters. To be specific, we calculate the mass of the backbone, the red bonds, and the shortest path. Moreover, we study key transport properties of percolation as represented by the random resistor network. We investigate the average two-point resistance as well as the entire family of multifractal moments of the current distribution. PMID- 14524855 TI - Microscopic theory of spatial-temporal congested traffic patterns at highway bottlenecks. AB - A microscopic theory of spatial-temporal congested traffic patterns at highway bottlenecks due to on-ramps, merge bottlenecks (a reduction of highway lanes), and off-ramps is presented. The basic postulate of three-phase traffic theory is used, which claims that homogeneous (in space and time) model solutions (steady states) of synchronized flow cover a two dimensional region in the flow-density plane [B. S. Kerner, Phys. Rev. Lett. 81, 3797 (1998); Trans. Res. Rec. 1678, 160 (1999)]. Phase transitions leading to diverse congested patterns, pattern evolution, and pattern nonlinear features have been found. Diagrams of congested patterns, i.e., regions of the pattern emergence dependent on traffic demand, have been derived. Diverse effects of metastability with respect to the pattern formation have been found. The microscopic theory allows us to explain the main empirical pattern features at on-ramps and off-ramps which have recently been found [B. S. Kerner, Phys. Rev. E 65, 046138 (2002)]. (i) Rather than moving jams, synchronized flow first occurs at bottlenecks if the flow rate is slowly increasing. Wide moving jams can spontaneously occur only in synchronized flow. (ii) General patterns (GP) and synchronized flow patterns (SP) can spontaneously emerge at the bottlenecks. There can be the widening SP (WSP), the moving SP (MSP), and the localized SP. (iii) At on-ramps cases of "weak" and "strong" congestion should be distinguished. In contrast to weak congestion, under strong congestion the flow rate in synchronized flow in GP reaches a limit flow rate, the frequency of the moving jam emergence reaches a maximum, i.e., the GP characteristics under strong congestion do not depend on traffic demand. (iv) At the off-ramp GP with weak congestion occur. (v) A study of the pattern formation on a highway with two bottlenecks shows that diverse expanded patterns can occur, which cover both bottlenecks. SP first emerged at the downstream bottleneck can be caught at the upstream bottleneck (the catch effect). MSP, WSP, or wide moving jams first emerged at the downstream bottleneck induce diverse patterns at the upstream bottleneck. The onset of congestion at the upstream bottleneck can lead to an intensification of congestion at the downstream bottleneck. This causes a change in the pattern type and/or the pattern features. PMID- 14524856 TI - Logarithmic corrections in dynamic isotropic percolation. AB - Based on the field theoretic formulation of the general epidemic process, we study logarithmic corrections to scaling in dynamic isotropic percolation at the upper critical dimension d=6. Employing renormalization group methods we determine these corrections for some of the most interesting time dependent observables in dynamic percolation at the critical point up to and including the next to leading correction. For clusters emanating from a local seed at the origin, we calculate the number of active sites, the radius of gyration, as well as the survival probability. PMID- 14524857 TI - Criticality and market efficiency in a simple realistic model of the stock market. AB - We discuss a simple model based on the minority game which reproduces the main stylized facts of anomalous fluctuations in finance. We present the analytic solution of the model in the thermodynamic limit. Stylized facts arise only close to a line of critical points with nontrivial properties, marking the transition to an unpredictable market. We show that the emergence of critical fluctuations close to the phase transition is governed by the interplay between the signal to noise ratio and the system size. These results provide a clear and consistent picture of financial markets, where stylized facts and verge of unpredictability are intimately related aspects of the same critical systems. PMID- 14524858 TI - Signal detection via residence times statistics: noise-mediated minimization of the measurement error. AB - We study the problem of detecting a small dc signal by quantifying its effect on the mean difference DeltaT in residence times in the stable steady states of a bistable dynamical measurement device, in the presence of a noise floor and a known time-sinusoidal bias signal. Errors in the measurement process occur due to a finite observation time that is present in most practical scenarios. The error is found to have a nonmonotonic dependence on the noise intensity; at a critical noise intensity, the error is minimized. This phenomenon, reminiscent of the well known stochastic resonance effect, can also be obtained by adjusting the device tuning parameters for a given noise floor. The effect appears to be most pronounced for subthreshold bias signals in the strongly nonlinear response regime. PMID- 14524859 TI - Order statistics of Rosenstock's trapping problem in disordered media. AB - The distribution of times t(j,N) elapsed until the first j independent random walkers from a set of N>>1, all starting from the same site, are trapped by a quenched configuration of traps randomly placed on a disordered lattice is investigated. In doing so, the cumulants of the distribution of the territory explored by N independent random walkers S(N)(t), and the probability Phi(N)(t) that no particle of an initial set of N is trapped by time t are considered. Simulation results for the two-dimensional incipient percolation aggregate show that the ratio between the nth cumulant and the nth moment of S(N)(t) is, for large N, (i) very large in comparison with the same ratio in Euclidean media, and (ii) almost constant. The first property implies that, in contrast with Euclidean media, approximations of the order higher than the standard zeroth-order Rosenstock approximation are required to provide a reasonable description of the trapping order statistics. Fortunately, the second property (which has a geometric origin) can be exploited to build these higher-order Rosenstock approximations. Simulation results for the two-dimensional incipient percolation aggregate confirm the predictions of our approach. PMID- 14524860 TI - Extended self-similarity in the two-dimensional metal-insulator transition. AB - We show that extended self-similarity, a scaling phenomenon first observed in classical turbulent flows, holds for a two-dimensional metal-insulator transition that belongs to the universality class of random Dirac fermions. Deviations from multifractality, which in turbulence are due to the dominance of diffusive processes at small scales, appear in the condensed-matter context as a large scale, finite-size effect related to the imposition of an infrared cutoff in the field theory formulation. We propose a phenomenological interpretation of extended self-similarity in the metal-insulator transition within the framework of the random beta-model description of multifractal sets. As a natural step, our discussion is bridged to the analysis of strange attractors, where crossovers between multifractal and nonmultifractal regimes are found and extended self similarity turns out to be verified as well. PMID- 14524861 TI - Three-dimensional randomly dilute Ising model: Monte Carlo results. AB - We perform a high-statistics simulation of the three-dimensional randomly dilute Ising model on cubic lattices L3 with L< or =256. We choose a particular value of the density, x=0.8, for which the leading scaling corrections are suppressed. We determine the critical exponents, obtaining nu=0.683(3), eta=0.035(2), beta=0.3535(17), and alpha=-0.049(9), in agreement with previous numerical simulations. We also estimate numerically the fixed-point values of the four point zero-momentum couplings that are used in field-theoretical fixed-dimension studies. Although these results somewhat differ from those obtained using perturbative field theory, the field-theoretical estimates of the critical exponents do not change significantly if the Monte Carlo result for the fixed point is used. Finally, we determine the six-point zero-momentum couplings, relevant for the small-magnetization expansion of the equation of state, and the invariant amplitude ratio R(+)(xi) that expresses the universality of the free energy density per correlation volume. We find R(+)(xi)=0.2885(15). PMID- 14524862 TI - Synchronization of vectorial noise-sustained structures. AB - The synchronization of vectorial, noise-sustained structures in nonlinear optical systems is discussed. In particular, the analysis is made for nondegenerate optical parametric oscillators with walk off. The interplay between walk off and noise fluctuations leads to the formation of noise-sustained transverse patterns in both the signal and idler fields. Despite the fact that both patterns are stochastic macroscopic structures driven by independent sources of noise, their correlation grows with time, finally leading to a spatially distributed time synchronization of noise-sustained structures. A physical explanation of this phenomenon is found by analyzing the linear instability process and the existence of exact nonlinear solutions that show the same correlation. PMID- 14524864 TI - Experimental observation of characteristic relations of type-III intermittency in the presence of noise in a simple electronic circuit. AB - We investigate the characteristic relations of type-II and -III intermittencies in the presence of noise. The theoretically predicted characteristic relation is that approximately exp[/epsilon/(2)] for a negative regime of epsilon and approximately epsilon(-nu) for the positive regime of epsilon (1/2 is the average laminar length and (1+epsilon) is the slope of the local Poincare map around the tangent point. We experimentally confirm these relations in a simple electronic circuit. PMID- 14524863 TI - Transition from spiral waves to defect-mediated turbulence induced by gradient effects in a reaction-diffusion system. AB - The transition from spiral waves to defect-mediated turbulence was studied in a spatial open reactor using Belousov-Zhabotinsky reaction. The experimental results show a new mechanism of the transition from spirals to spatiotemporal chaos, in which the gradient effects in the three-dimensional system are essential. The transition scenario consists of two stages: first, the effects of gradients in the third dimension cause a splitting of the spiral tip and a deletion of certain wave segments, generating new wave sources; second, the waves sent by the new wave sources undergo a backfire instability, and the back waves are laterally unstable. As a result, defects are automatically generated and fill all over the system. The result of numerical simulation using the FitzHugh-Nagumo model essentially agrees with the experimental observation. PMID- 14524865 TI - Low-frequency fluctuations in the Lang-Kobayashi equations. AB - The Lang-Kobayashi equations are simplified by a local analysis that focuses, in the long-delay-time limit, on one pair of mode-antimode only. In the domain of hysteresis between the two steady states, low frequency fluctuations (LFF) can be observed if there is a domain of bistability where both steady states are unstable. The high-frequency oscillations and the drop-offs in the LFF regime are associated with a dynamics close to the unstable upper and lower branch steady states, respectively. PMID- 14524866 TI - Experiments on elastomechanical wave functions in chaotic plates and their statistical features. AB - We measure the amplitude of the elastomechanical displacement at a fine grid of points on a free plate having the shape of a Sinai stadium. The obtained displacement field formally corresponds to a wave function in a quantum system. While the distribution of the squared amplitudes agrees with the prediction of random matrix theory (RMT), there is a strong deviation of the spatial correlator from the standard prediction for quantum chaotic systems. We show that this is due to the presence of two modes, leading to a beating phenomenon. We construct a proper extension of the spatial correlator within the framework of RMT. PMID- 14524867 TI - Stability domains for time-delay feedback control with latency. AB - We generalize a known analytical method for determining the stability of periodic orbits controlled by time-delay feedback methods when latencies associated with the generation and injection of the feedback signal cannot be ignored. We discuss the case of extended time-delay autosynchronization and show that nontrivial qualitative features of the domain of control observed in experiments can be explained by taking into account the effects of both the unstable eigenmode and a single stable eigenmode in the Floquet theory. PMID- 14524868 TI - Asymptotic solutions of stationary patterns in convection-reaction-diffusion systems. AB - We study and map the possible stationary patterns that emerge in a convection reaction-diffusion (CRD) system using a learning polynomial kinetics. We classify the patterns according to the kinetic model (oscillatory, bistable, or intermediate), the instability nature of the bounded system (convective or absolute), the applied boundary conditions and the system length. This analysis presents a unifying approach to various pattern-inducing mechanisms such as DIFICI (differential flow induced chemical instability), which predicts moving patterns in systems with wide difference of convective rates, and differential capacity patterns, which predicts stationary patterns in cross-flow reactors with a large heat capacity. Previous studies of CRD systems have considered only oscillatory kinetics. Nonlinear analysis, which follows the front motion by approximating its velocity, accounts for the stability of the stationary, whether spatially periodic or other, patterns. The most dominant state is the large amplitude stationary spatially periodic pattern. With oscillatory kinetics these emerge in the convectively unstable domain above the amplification threshold. The domain of absolute instability, which is determined analytically for unbounded systems, is divided in the bounded system into two subdomains with moving DIFICI waves or stationary patterns. With bistable kinetics the large-amplitude stationary patterns can be sustained only within a narrow subdomain but other stationary patterns, that incorporate several fronts upstream and an "almost homogeneous" tail downstream, can be sustained as well. With intermediate kinetics the large-amplitude axisymmetric stationary patterns may coexist with small-amplitude stationary nonaxisymmetric patterns. PMID- 14524869 TI - Complete synchronization and generalized synchronization of one-way coupled time delay systems. AB - The complete synchronization and generalized synchronization (GS) of one-way coupled time-delay systems are studied. We find that GS can be achieved by a single scalar signal, and its synchronization threshold for different delay times shows the parameter resonance effect, i.e., we can obtain stable synchronization at a smaller coupling if the delay time of the driven system is chosen such that it is in resonance with the driving system. Near chaos synchronization, the desynchronization dynamics displays periodic bursts with the period equal to the delay time of the driven system. These features can be easily applied to the recovery of time-delay systems. PMID- 14524870 TI - Experimental evidence of van der Pol-Fitzhugh-Nagumo dynamics in semiconductor optical amplifiers. AB - Thermo-optical pulsing in semiconductor amplifiers is experimentally shown to correspond to a very common excitable scenario (the van der Pol-Fitzhugh-Nagumo system). Self-sustained oscillations appear in the sequence predicted by this simple dynamical model as we change either the injection level or the bias current. Periodic modulation of these parameters leads to the characteristic phase-locking structure. Furthermore, coherence resonance is observed when external noise is added to the system. PMID- 14524871 TI - Isothermal flame balls: effect of autocatalyst decay. AB - The steady, spherically symmetric solutions to the reaction-diffusion equations based on a simple autocatalytic reaction followed by the decay of the autocatalyst are considered. Three parameters-the orders with respect to the autocatalyst in the autocatalysis p and in the decay q and the rate of decay of the autocatalyst relative to its autocatalytic production K-determine the steady concentration profiles. Numerical integrations for a fixed value of the order of the autocatalyst show that the concentration profiles have different forms depending on whether q

/=p. In the former case, there is a critical decay rate K(crit) for solutions to exist, with multiple solutions for K3, 6-->3, 5-->3, and 4-->3 transitions in Ni- to Kr like gold ions produced in the Livermore electron beam ion traps EBIT-I and EBIT II has been recorded with an x-ray crystal spectrometer and a photometrically calibrated microcalorimeter. The plasmas had either monoenergetic electron beams with E(beam)=2.66, 3.53, or 4.54 keV or an experimentally simulated thermal electron distributions with T(e)=2.5 keV. The electron densities were approximately 10(12)cm(-3). The measured spectra have been compared to atomic structure calculations and synthetic spectra provided by the Hebrew University Lawrence Livermore Atomic Code atomic data package. Line identifications and accurate photon energy measurements have been made for many collisionally excited transitions. Approximately 140 lines have been identified in nine charge states. Agreement within 20-30 % exists between the measured and modeled line intensities for most lines excited by the monoenergetic electron beam plasmas, although some larger discrepancies can be found for some weaker features. PMID- 14524899 TI - Frequency-dependent reflectivity of shock-compressed xenon plasmas. AB - Results for the reflection coefficient of shock-compressed dense xenon plasmas at pressures of 1.6-20 GPa and temperatures around 30 000 K using laser beams of wavelengths 1.06 micro m and 0.694 micro m are presented, which indicate metallic behavior at high densities. For the theoretical description of the experiments, a quantum statistical approach to the dielectric function is used. The comparison with molecular dynamics simulations is discussed. We conclude that reflectivity measurements at different wavelengths can provide information about the density profile of the shock wave front. PMID- 14524901 TI - Coulomb explosions and energy loss of molecular ions in plasmas. AB - Interactions of swift molecular ions with high-density plasma targets are studied by means of the linearized Vlasov-Poisson theory, allowing the dynamically screened interaction potential among the constituent ions to be expressed in terms of the classical plasma dielectric function. Coulomb explosions and the energy losses of a molecular ion are simulated by solving the equations of motion for the constituent ions. It is found that, due to the wakelike asymmetry of the interaction potential, the molecular axis tends to align itself along the beam direction. In addition, a strong enhancement of the energy loss of the molecular ion has been found in the initial stages of Coulomb explosions due to proximity of the constituent ions, but this effect diminishes at latter stages when the ions are sufficiently far apart. PMID- 14524900 TI - Excitation of ion-wave wakefield by the resonant absorption of a short pulsed microwave with plasma. AB - Unmagnetized, inhomogeneous laboratory plasma irradiated by a high power (eta=E(2)(0)/4pin(e)kT(e) approximately 5.0x10(-2)) short pulsed microwave with pulse length of the order of ion-plasma period (tau(pi) less, similar 2pi/omega(pi)) is studied. Large density perturbation traveling through the underdense plasma with a velocity much greater than the ion sound speed produced by the resonant absorption of the microwave pulse has been observed. In the beginning the density perturbation has large amplitude (deltan/n(0) approximately 40%) and propagates with a velocity of the order of 10(6) cm/s. But later its amplitude as well as the velocity decrease rapidly, and finally the velocity arrives with twice the ion sound speed. The oscillating incident electromagnetic waves enhance highly localized electric field by the resonant absorption process and develop time-averaged force field which pushes plasma electrons from the resonant layer. As the electrons are accelerated to be ejected, they pull plasma ions as a bunch with them by means of self-consistent Coulomb force. This suprathermal ion bunch can excite an ion-wave wakefield. PMID- 14524902 TI - Method of measuring the coupled lattice functions at the interaction point in e+e storage rings. AB - We have investigated a method of measuring the complete lattice functions including the coupling parameters at any azimuthal position in a periodic and symplectic system. In particular, the method is applied to measure the lattice functions at the interaction point where the beams collide. It has been demonstrated that a complete set of lattice functions can be accurately measured with two adjacent beam position monitors and the known transformation matrix between them. As a by-product, the method also automatically measures the complete one-turn matrix. PMID- 14524903 TI - Wake field in dielectric acceleration structures. AB - In this study we present a general approach for the analysis of the wake field of a point charge moving in a vacuum tunnel bored in dielectric material that is uniform in the direction parallel to the motion of the bunch. In the transverse direction the structure surrounding the dielectric may have arbitrary geometry. A quasianalytic expression that relates the decelerating force with the first dielectric layer, the radius of the vacuum tunnel where the charge moves, and the reflection characteristics of the structure has been developed. Simulation results for a simple structure indicate that, if the effective location where the reflection occurs in the dielectric is sufficiently apart from the edge of the vacuum tunnel, it has no effect on the point charge. In fact, the decelerating field converges exponentially as this distance increases, to the asymptotic value determined by the first dielectric layer. An estimate of the trailing wake when the structure supports a specific mode is also provided. PMID- 14524904 TI - Halo formation at early stage of injection in high-intensity hadron rings. AB - Halo formation under a nonequilibrium state for a two-dimensional Gaussian beam in a FODO lattice, which is an array of magnets where F is focusing, D is defocusing, and O is the drift space between magnets, was examined in terms of a transition of time-varying nonlinear resonances. Nonlinear resonant-interactions between individual particles and intrinsic beam-core oscillations result in a beam halo. The location of the halo is analytically tractable using canonical equations derived from an isolated resonance Hamiltonian. PMID- 14524905 TI - X rays from relativistic electrons in a multilayer structure. AB - A dynamic diffraction theory of x-ray emission by relativistic electrons crossing a finite-thickness multilayer mirror (e.g., alternating layers of W and B4C) is developed, taking into account both diffracted transition and parametric radiation mechanisms. Simple formulas describing the characteristics of the total emission from either thin nonabsorbing or thick absorbing multilayers are derived. These formulas show that a multilayer radiator can be brighter and more efficient than crystalline ones. Good agreement between theory and prior experimental results is also shown. Thus the theory and its experimental verification demonstrate the possibility of a tunable quasimonochromatic x-ray source whose efficiency can be larger than that of other novel x-ray sources. PMID- 14524906 TI - Dynamical instabilities of quasistatic crack propagation under thermal stress. AB - We address the theory of quasistatic crack propagation in a strip of glass that is pulled from a hot oven towards a cold bath. This problem had been carefully studied in a number of experiments that offer a wealth of data to challenge the theory. We improve upon previous theoretical treatments in a number of ways. First, we offer a technical improvement of the discussion of the instability towards the creation of a straight crack. This improvement consists in employing Pade approximants to solve the relevant Wiener-Hopf factorization problem that is associated with this transition. Next we improve the discussion of the onset of oscillatory instability towards an undulating crack. We offer a way of considering the problem as a sum of solutions of a finite strip without a crack and an infinite medium with a crack. This allows us to present a closed form solution of the stress intensity factors in the vicinity of the oscillatory instability. Most importantly we develop a dynamical description of the actual trajectory in the regime of oscillatory crack. This theory is based on the dynamical law for crack propagation proposed by Hodgdon and Sethna. We show that this dynamical law results in a solution of the actual crack trajectory in post critical conditions; we can compute from first principles the critical value of the control parameters, and the characteristics of the solution such as the wavelength of the oscillations. We present detailed comparison with experimental measurements without any free parameters. The comparison appears quite excellent. Finally we show that the dynamical law can be translated to an equation for the amplitude of the oscillatory crack; this equation predicts correctly the scaling exponents observed in experiments. PMID- 14524907 TI - Stochastic resonance in the presence of spatially localized structures. AB - Stable spatially localized structures exist in a wide variety of spatially extended nonlinear systems, including nonlinear optical devices. We study stochastic resonance (SR) in models of optical parametric oscillators in the presence of a spatially uniform time-periodic driving and in a regime where two equivalent states with equal intensity but opposite phase exist. Diffraction and nonlinearity enable the existence of localized states, formed by the locking of kinks and antikinks and displaying spatially damped oscillatory tails (in one dimension) or the stabilization of dark ring cavity solitons (in two dimensions). We show that SR is inhibited at low driving amplitudes by the presence of localized states which obstruct the front motion. For larger driving amplitudes, in the regime where localized states cease to be stable, we observe instead an enhancement of SR. PMID- 14524908 TI - Linear dynamics of double-porosity dual-permeability materials. I. Governing equations and acoustic attenuation. AB - The equations governing the linear acoustics of composites with two isotropic porous constituents are derived from first principles using volume-averaging arguments. The theory is designed for modeling acoustic propagation through heterogeneous porous structures. The only restriction placed on the geometry of the two porous phases is that the overall composite remains isotropic. The theory determines the macroscopic fluid response in each porous phase in addition to the combined bulk response of the grains and fluid in the composite. The complex frequency-dependent macroscopic compressibility laws that are obtained allow for fluid transfer between the porous constituents. Such mesoscopic fluid transport between constituents within each averaging volume provides a distinct attenuation mechanism from the losses associated with the net Darcy flux within individual constituents as is quantified in the examples. PMID- 14524909 TI - Linear dynamics of double-porosity dual-permeability materials. II. Fluid transport equations. AB - For the purpose of understanding the acoustic attenuation of double-porosity composites, the key macroscopic equations are those controlling the fluid transport. Two types of fluid transport are present in double-porosity dual permeability materials: (1) a scalar transport that occurs entirely within each averaging volume and that accounts for the rate at which fluid is exchanged between porous phase 1 and porous phase 2 when there is a difference in the average fluid pressure between the two phases and (2) a vector transport that accounts for fluid flux across an averaging region when there are macroscopic fluid-pressure gradients present. The scalar transport that occurs between the two phases can produce large amounts of wave-induced attenuation. The scalar transport equation is derived using volume-averaging arguments and the frequency dependence of the transport coefficient is obtained. The dual-permeability vector Darcy law that is obtained allows for fluid flux across each phase individually and is shown to have a symmetric permeability matrix. The nature of the cross coupling between the flow in each phase is also discussed. PMID- 14524910 TI - Dynamics of multimode diode lasers with strong, frequency-selective optical feedback. AB - The dynamical behavior of a class of multimode semiconductor diode lasers with emission wavelength around 980 nm is investigated both experimentally and numerically in the presence of strong, frequency-selective optical feedback provided by a fiber Bragg grating. The focus is set on the switching between broad- and narrow-band optical spectra, on chaotic transitions, and on the loss of frequency locking between laser and grating. Laser and feedback parameters are chosen in the typical ranges pertaining to wavelength stabilization in erbium doped fiber amplifiers for telecommunication applications. An improved set of rate equations, which allows for arbitrary feedback levels and includes experimentally measured gain and linewidth enhancement factor, is studied analytically and numerically. PMID- 14524911 TI - Standard and embedded solitons in nematic optical fibers. AB - A model for a non-Kerr cylindrical nematic fiber is presented. We use the multiple scales method to show the possibility of constructing different kinds of wave packets of transverse magnetic modes propagating through the fiber. This procedure allows us to generate different hierarchies of nonlinear partial differential equations which describe the propagation of optical pulses along the fiber. We go beyond the usual weakly nonlinear limit of a Kerr medium and derive a complex modified Korteweg-de Vries equation (CM KdV) which governs the dynamics for the amplitude of the wave packet. In this derivation the dispersion, self focussing, and diffraction in the nematic fiber are taken into account. It is shown that this CM KdV equation has two-parameter families of bright and dark complex solitons. We show analytically that under certain conditions, the bright solitons are actually double-embedded solitons. We explain why these solitons do not radiate at all, even though their wave numbers are contained in the linear spectrum of the system. We study (numerically and analytically) the stability of these solitons. Our results show that these embedded solitons are stable solutions, which is an interesting property since in most systems the embedded solitons are weakly unstable solutions. Finally, we close the paper by making comments on the advantages as well as the limitations of our approach, and on further generalizations of the model and method presented. PMID- 14524912 TI - Incoherent white light solitons in logarithmically saturable noninstantaneous nonlinear media. AB - We analytically demonstrate the existence of white light solitons in logarithmically saturable noninstantaneous nonlinear media. This incoherent soliton has elliptic Gaussian intensity profile, and elliptic Gaussian spatial correlation statistics. The existence curve of the soliton connects the strength of the nonlinearity, the spatial correlation distance as a function of frequency, and the characteristic width of the soliton. For this soliton to exist, the spatial correlation distance must be smaller for larger temporal frequency constituents of the beam. PMID- 14524913 TI - Radiation from a dipole embedded in a multilayer slab. AB - An analytical solution for the radiation emitted from a dipole embedded in an arbitrary, planar dielectric film stack is presented. The calculation uses a rigorous Hertz-vector formalism to treat the electromagnetic boundary conditions. The radiation fields are then evaluated in a far-field approximation to get the radiated fields far from the dipole. Both two-dimensional (2D) emission into bound modes of the dielectric stack and three-dimensional (3D) emission into radiation fields above and below the stack are evaluated. These solutions are explored for two simple cases: a InGaAs slab symmetrically clad with up to four high-contrast (Al(2)O(3)/GaAs) Bragg mirror pairs and semi-infinite air spaces, and a similar asymmetric structure with a GaAs substrate on one side. The symmetric structure supports both 2D bound and 3D radiation fields. The asymmetric structure only supports 3D radiation fields since there are no strictly bound modes, but "leaky" modes appear that are very similar to the bound modes in the symmetric structure except that the radiated power ultimately is transmitted into the substrate in a very highly directional beam. This calculation is applicable to a wide range of solid-state photonic devices, including vertical-cavity and edge-emitting lasers, spontaneous light-emitting diodes, and photodetectors. PMID- 14524914 TI - Oblique frozen modes in periodic layered media. AB - We study the classical scattering problem of a plane electromagnetic wave incident on the surface of semi-infinite periodic stratified media incorporating anisotropic dielectric layers with special oblique orientation of the anisotropy axes. We demonstrate that an obliquely incident light, upon entering the periodic slab, gets converted into an abnormal grazing mode with huge amplitude and zero normal component of the group velocity. This mode cannot be represented as a superposition of extended and evanescent contributions. Instead, it is related to a general (non-Bloch) Floquet eigenmode with the amplitude diverging linearly with the distance from the slab boundary. Remarkably, the slab reflectivity in such a situation can be very low, which means an almost 100% conversion of the incident light into the axially frozen mode with the electromagnetic energy density exceeding that of the incident wave by several orders of magnitude. The effect can be realized at any desirable frequency, including optical and UV frequency range. The only essential physical requirement is the presence of dielectric layers with proper oblique orientation of the anisotropy axes. Some practical aspects of this phenomenon are considered. PMID- 14524915 TI - Conservative form of the density of states of a photonic crystal with a pseudogap. AB - We show that the total number of states in a photonic crystal in the entire allowed frequency regime will be conserved, and it is equal to that of its corresponding effective medium, i.e., if the density of states (DOS) has a valley(s) in some range(s) of frequencies, it must be compensated for by increases over some other range(s). This rule is of importance in developing a model pseudogap in order to describe the mean emission characteristics of the system when there is a collection of dependently emitting atoms or molecules with essentially random dipole orientations in a large volume and the spectrum of the active atoms is wide enough. This is because, with this rule, the states conservative model always results in DOS-induced suppression, absolute enhancement, narrowing, spectrum splits, and redshift or blueshift of spontaneous emission spectra. PMID- 14524916 TI - Variational analysis for photonic molecules: application to photonic benzene waveguides. AB - A type of artificial molecule, called the photonic molecule, is investigated. It consists of coupled-defect atoms in photonic crystals. We theoretically present a method to determine the frequency of each resonant mode for the photonic molecule. Within the major band gap, the photonic molecule confines the resonant modes that are closely analogous to the ground-state molecular orbitals of an analogous chemical molecule and the corresponding electromagnetic spectrum is also isomorphous to the electric energy levels. By employing the variational theory, the constraint determining the resonant coupling is formulated, which is consistent with the results of both the scattering matrix method and the group analysis. In addition, a type of photonic waveguide is proposed for the two dimensional system that manipulates the mechanism of photon hopping between photonic benzene and offers an optical feature of twin waveguiding bandwidths. PMID- 14524917 TI - Using the finite domain remnant of the continuous spectrum to examine integrability: effect of boundary conditions. AB - The aim of this work is to propose a method for testing the integrability of a model partial differential (PDE) and/or differential difference equation (DDE), by examining it in a finite but large domain. For monoparametric families of PDE/DDE's, that are known to possess isolated integrable points, we find that very special features occur in the finite domain remnant of the continuous ("phonon") spectrum at these "singular" points. We identify these features in the case example of a PDE and a DDE (that sustain front and pulselike solutions, respectively) for different types of boundary conditions. The key finding of the work is that such spectral features are generic near the singular, integrable points and hence we propose to explore a given PDE/DDE in a finite but large domain for such traits, as a means of assessing its potential integrability. PMID- 14524918 TI - Solutions for vectorial beam coupling under ac field in cubic photorefractive crystals. AB - We develop a theory of vectorial wave coupling in cubic photorefractive crystals placed in an alternating ac field to enhance the nonlinear response. It is proven in the general case that despite essential differences between the diffusion and the ac nonlocal responses, the latter keeps the light interference fringes straight during the interaction. This fundamental feature allows, under certain restrictions, to reduce the nonlinear problem of vectorial coupling to the known linear problem of vectorial Bragg diffraction from a spatially uniform grating, which admits an exact solution. As a result, the nonlinear vectorial problem can be effectively solved for a number of practically important cases. These cases include nontrivial polarization effects and also the high-contrast effects. The presence of conservation laws involving the polarization degrees of freedom is shown. A number of particular cases relevant to experiments with BTO crystals are considered. PMID- 14524919 TI - Quadratic solitons as nonlocal solitons. AB - We show that quadratic solitons are equivalent to solitons of a nonlocal Kerr medium. This provides new physical insight into the properties of quadratic solitons, often believed to be equivalent to solitons of an effective saturable Kerr medium. The nonlocal analogy also allows for analytical solutions and the prediction of bound states of quadratic solitons. PMID- 14524920 TI - Transport coefficients for stochastic rotation dynamics in three dimensions. AB - Explicit expressions for the transport coefficients of a recently introduced stochastic model for simulating fluctuating fluid dynamics are derived in three dimensions by means of Green-Kubo relations and simple kinetic arguments. The results are shown to be in excellent agreement with simulation data. Two collision rules are considered and their computational efficiency is compared. PMID- 14524921 TI - Polynomial iterative algorithms for coloring and analyzing random graphs. AB - We study the graph coloring problem over random graphs of finite average connectivity c. Given a number q of available colors, we find that graphs with low connectivity admit almost always a proper coloring whereas graphs with high connectivity are uncolorable. Depending on q, we find with a one-step replica symmetry breaking approximation the precise value of the critical average connectivity c(q). Moreover, we show that below c(q) there exists a clustering phase c in [c(d),c(q)] in which ground states spontaneously divide into an exponential number of clusters. Furthermore, we extended our considerations to the case of single instances showing consistent results. This leads us to propose a different algorithm that is able to color in polynomial time random graphs in the hard but colorable region, i.e., when c in [c(d),c(q)]. PMID- 14524922 TI - Characterization of the probabilistic traveling salesman problem. AB - We show that stochastic annealing can be successfully applied to gain new results on the probabilistic traveling salesman problem. The probabilistic "traveling salesman" must decide on an a priori order in which to visit n cities (randomly distributed over a unit square) before learning that some cities can be omitted. We find the optimized average length of the pruned tour follows E(L(pruned))=sqrt[np](0.872-0.105p)f(np), where p is the probability of a city needing to be visited, and f(np)-->1 as np--> infinity. The average length of the a priori tour (before omitting any cities) is found to follow E(L(a priori))=sqrt[n/p]beta(p), where beta(p)=1/[1.25-0.82 ln(p)] is measured for 0.05< or =p< or =0.6. Scaling arguments and indirect measurements suggest that beta(p) tends towards a constant for p<0.03. Our stochastic annealing algorithm is based on limited sampling of the pruned tour lengths, exploiting the sampling error to provide the analog of thermal fluctuations in simulated (thermal) annealing. The method has general application to the optimization of functions whose cost to evaluate rises with the precision required. PMID- 14524923 TI - Boundary conditions for stochastic solutions of the convection-diffusion equation. AB - Stochastic methods offer an attractively simple solution to complex transport controlled problems, and have a wide range of physical, chemical, and biological applications. Stochastic methods do not suffer from the numerical diffusion that plagues grid-based methods, but they typically lose accuracy in the vicinity of interfacial boundaries. In this work we introduce some ideas and algorithms that can be used to implement boundary conditions in stochastic simulations of the convection-diffusion equation with accuracies comparable to the bulk phase. The algorithms have been tested in two-dimensional channel flows over a range of Peclet numbers, and compared with independent finite-difference calculations. PMID- 14524924 TI - Can a numerically stable subgrid-scale model for turbulent flow computation be ideally accurate?: a preliminary theoretical study for the Gaussian filtered Navier-Stokes equations. AB - This paper introduces a candidate for the origin of the numerical instabilities in large eddy simulation repeatedly observed in academic and practical industrial flow computations. Without resorting to any subgrid-scale modeling, but based on a simple assumption regarding the streamwise component of flow velocity, it is shown theoretically that in a channel-flow computation, the application of the Gaussian filtering to the incompressible Navier-Stokes equations yields a numerically unstable term, a cross-derivative term, which is similar to one appearing in the Gaussian filtered Vlasov equation derived by Klimas [J. Comput. Phys. 68, 202 (1987)] and also to one derived recently by Kobayashi and Shimomura [Phys. Fluids 15, L29 (2003)] from the tensor-diffusivity subgrid-scale term in a dynamic mixed model. The present result predicts that not only the numerical methods and the subgrid-scale models employed but also only the applied filtering process can be a seed of this numerical instability. An investigation concerning the relationship between the turbulent energy scattering and the unstable term shows that the instability of the term does not necessarily represent the backscatter of kinetic energy which has been considered a possible origin of numerical instabilities in large eddy simulation. The present findings raise the question whether a numerically stable subgrid-scale model can be ideally accurate. PMID- 14524925 TI - Theory of the lattice Boltzmann method: acoustic and thermal properties in two and three dimensions. AB - The focus of the present work is to provide an analysis for the acoustic and thermal properties of the energy-conserving lattice Boltzmann models, and a solution to the numerical defects and instability associated with these models in two and three dimensions. We discover that a spurious algebraic coupling between the shear and energy modes of the linearized evolution operator is a defect universal to the energy-conserving Boltzmann models in two and three dimensions. This spurious mode coupling is highly anisotropic and may occur at small values of wave number k along certain directions, and it is a direct consequence of the following key features of the lattice Boltzmann equation: (1) its simple spatial temporal dynamics, (2) the linearity of the relaxation modeling for collision operator, and (3) the energy-conservation constraint. To eliminate the spurious mode coupling, we propose a hybrid thermal lattice Boltzmann equation (HTLBE) in which the mass and momentum conservation equations are solved by using the multiple-relaxation-time model due to d'Humieres, whereas the diffusion-advection equation for the temperature is solved separately by using finite-difference technique (or other means). Through the Chapman-Enskog analysis we show that the hydrodynamic equations derived from the proposed HTLBE model include the equivalent effect of gamma=C(P)/C(V) in both the speed and attenuation of sound. Appropriate coupling between the energy and velocity field is introduced to attain correct acoustics in the model. The numerical stability of the HTLBE scheme is analyzed by solving the dispersion equation of the linearized collision operator. We find that the numerical stability of the lattice Boltzmann scheme improves drastically once the spurious mode coupling is removed. It is shown that the HTLBE scheme is far superior to the existing thermal LBE schemes in terms of numerical stability, flexibility, and possible generalization for complex fluids. We also present the simulation results of the convective flow in a rectangular cavity with different temperatures on two opposite vertical walls and under the influence of gravity. Our numerical results agree well with the pseudospectral result. PMID- 14524926 TI - Errorless reproduction of given pattern dynamics by means of cellular automata. AB - In this paper we propose the two methods to reproduce given binary pattern dynamics with cellular automata. The point is that one can easily find a sequence of rules or specified rules in two-state multineighbors cellular automata, which enable an errorless description and reproduction of given multiple sequences of binary patterns. Actual examples using computer experiments for one-dimensional bit-pattern data (digital sound signals, multiple sequences of cycle patterns) are given. Noise robustness and the other important dynamical properties of these methods are investigated from the perspective of "rule dynamics" and in comparison with a recurrent neural network model, which enables us to embed given binary patterns as multiple attractors in the form of fixed points or limit cycles. PMID- 14524927 TI - Strange nonchaotic attractors in driven excitable systems. AB - Through quasiperiodic forcing, an excitable system can be driven into a regime of spiking behavior that is both aperiodic and stable. This is a consequence of strange nonchaotic dynamics: the motion of the system is on a fractal attractor and the largest Lyapunov exponent is negative. PMID- 14524928 TI - Unexpected correspondence between noise-induced and master-slave complete synchronizations. AB - A comparison between noise-induced synchronization and master-slave (Pecora Carroll) synchronization is investigated in this paper. We find an interesting correspondence between the effective driving variables of these two kinds of synchronizations in three-dimensional chaotic systems, when the systems have nonlinear terms in more than one equation. A study of the Lorenz model, the Hindmarsh-Rose neuron model, and the Hastings-Powell foodweb model is given to support this claim. It is a somewhat surprising result since these two kinds of synchronizations arise through different mechanisms. We also examine an exceptional case, where the nonlinear term of the system appears in a single equation, as in the Pikovsky-Rabinovich circuit model, and explain why the correspondence fails. PMID- 14524929 TI - Band structures and band gaps of liquid surface waves propagating through an infinite array of cylinders. AB - The multiple scattering method is applied to the calculations of band structures of liquid surface waves propagating through an infinite array of vertical cylinders. The influence of the filling fraction on the formation of band gaps is discussed. It is found that there exist complete band gaps for both the square and triangular arrays of cylinders. PMID- 14524930 TI - Surface instabilities on liquid oxygen in an inhomogeneous magnetic field. AB - Liquid oxygen exhibits surface instabilities when subjected to a sufficiently strong magnetic field. A vertically oriented magnetic field gradient both increases the magnetic field value at which the pattern forms and shrinks the length scale of the surface patterning. We show that these effects of the field gradient may be described in terms of an "effective gravity," which in our experiments may be varied from 1 g to 360 g. PMID- 14524931 TI - Effect of liquid films on the isothermal drying of porous media. AB - We study the effects of liquid films on the isothermal drying of porous media. They are important for the transport of liquid to an evaporation interface, far from the receding liquid clusters. Through a transformation, the drying problem is mapped to the Laplace equation around the percolation liquid clusters. From its solution, the properties of drying are obtained in terms of the capillary number. Consistent with experimental evidence, film flow is shown to accelerate drying significantly. PMID- 14524932 TI - Magnetic-field-limited currents. AB - An upper limit on the net current of a charged particle beam is derived by requiring the energy per unit length in the magnetic field to be less than that in the particles. The limit is calculated for five different current profiles. It is shown that an arbitrarily large net current can propagate in a large diameter ring. The results are found to be closely related to the Alfven limit. A limit on the forward current in a conductor is calculated, which defines a magnetic inhibition time. PMID- 14524933 TI - Higher-order neural networks, Polya polynomials, and Fermi cluster diagrams. AB - The problem of controlling higher-order interactions in neural networks is addressed with techniques commonly applied in the cluster analysis of quantum many-particle systems. For multineuron synaptic weights chosen according to a straightforward extension of the standard Hebbian learning rule, we show that higher-order contributions to the stimulus felt by a given neuron can be readily evaluated via Polya's combinatoric group-theoretical approach or equivalently by exploiting a precise formal analogy with fermion diagrammatics. PMID- 14524934 TI - Computational approach for modeling intra- and extracellular dynamics. AB - We introduce a phase-field approach for diffusion inside and outside a closed cell with damping and with source terms at the stationary interface. The method is compared to exact solutions (where possible) and the more traditional finite element method. It is shown to be very accurate, easy to implement, and computationally inexpensive. We apply our method to a recently introduced model for chemotaxis by Rappel et al. [Biophys. J. 83, 1361 (2002)]. PMID- 14524935 TI - Structure optimization in an off-lattice protein model. AB - We study an off-lattice protein toy model with two species of monomers interacting through modified Lennard-Jones interactions. Low energy configurations are optimized using the pruned-enriched-Rosenbluth method (PERM), hitherto employed to native state searches only for off-lattice models. For two dimensions we found states with lower energy than previously proposed putative ground states for all chain lengths >/=13. This indicates that PERM has the potential to produce native states also for more realistic protein models. For d=3, where no published ground states exist, we present some putative lowest energy states for future comparison with other methods. PMID- 14524936 TI - Comment on "Ehrenfest times for classically chaotic systems". AB - In a recent Rapid Communication [P. G. Silvestrov and C. W. J. Beenakker, Phys. Rev. E 65, 035208(R) (2002)], the authors, Silvestrov and Beenakker, introduce a way to lengthen the Ehrenfest time tau for fully chaotic systems. We disagree with several statements made in their paper, and address the following points essential to their conclusions: (1) it is not true that all semiclassical approximations for chaotic systems fail at a so-called "log time" tau proportional, variant -ln( variant Planck's over 2pi ), differing only by a numerical coefficient; and (2) the limitation of the semiclassical approximation as expressed in the authors' Eq. (8) is not limited by their argument leading to Eq. (12). PMID- 14524938 TI - Experimental study of noise-induced phase synchronization in vertical-cavity lasers. AB - We report the experimental evidence of noise-induced phase synchronization in a vertical-cavity laser. The polarized laser emission is entrained with the input periodic pump modulation when an optimal amount of white, Gaussian noise is applied. We characterize the phenomenon, evaluating the average frequency of the output signal and the diffusion coefficient of the phase difference variable. Their values are roughly independent of the different wave forms of periodic input, provided that a simple condition for the amplitudes is satisfied. The experimental results are compared with numerical simulations of a Langevin model. PMID- 14524939 TI - Analysis by x-ray microtomography of a granular packing undergoing compaction. AB - Several acquisitions of x-ray microtomography have been performed on a beads packing while it compacts under vertical vibrations. An image analysis allows to study the evolution of the packing structure during its progressive densification. In particular, the volume distribution of the pores reveals a large tail, compatible to an exponential law, which slowly reduces as the system gets more compact. This is quite consistent, for large pores, with the free volume theory. These results are also in very good agreement with those obtained by a previous numerical model of granular compaction. PMID- 14524940 TI - Diffusion-limited aggregation in channel geometry. AB - We performed extensive numerical simulation of diffusion-limited aggregation in two-dimensional channel geometry. Contrary to earlier claims, the measured fractal dimension D=1.712+/-0.002 and its leading correction to scaling are the same as in the radial case. The average cluster, defined as the average conformal map, is similar but not identical to Saffman-Taylor fingers. PMID- 14524941 TI - Ultrafast laser-induced molecular and morphological changes during spinodal demixing of water/2-butoxyethanol/KCl. AB - We initiated morphological and molecular level changes in the spinodal decomposition (SD) of H(2)O/2-butoxyethanol/KCl with a pulsed ir laser. Transient Raman spectra gave us a molecular level view of the early stage of this process that could be linked to later morphological events. Chemical changes during SD, such as reorganization of H bonds and forced hydrophobic interactions, ended after 1 micros; however, phase domains continued to grow with self-similarity after 30 micros. The growth of the phase domains satisfied the power law L(t) approximately t(0.55) and was consistent with the late stage of SD. The time scale for the onset of late stage SD is many orders of magnitude faster than previously reported in ionic and nonionic conditions. PMID- 14524943 TI - Reabsorption effect and spatial ordering in island ripening. AB - A model for detachment-limited island ripening is analyzed in terms of a reabsorption (or pedophagous) effect where islands releasing monomers can reabsorb their own offspring. When the effect is artificially turned off, ripened island arrays are spatially disordered and the island growth rates and size distribution are easily understood in terms of a classic mean field theory. With the pedophagous effect included, the arrays develop a spatial ordering consistent with experimental studies. The ordering is quantified through a correlation between island size and the number of nearby neighbors, yielding modified mean field island growth rates and a much-improved expression for the island size distribution. It is suggested that the spatial ordering and our explanation are widely applicable to ripening systems. PMID- 14524942 TI - Interrupted coarsening of anisotropic step meander. AB - We report on the effect of anisotropy on the step meandering instability on vicinal surfaces during molecular beam epitaxy growth. A scenario of interrupted coarsening is found: the lateral length scale of the structure first significantly increases with time and then freezes at a larger length scale. The wavelength selection mechanism results from a nontrivial nonlinear effect of anisotropy. Anisotropy also leads to solutions which drift sideways, resulting from the loss of the back-front symmetry of the meander and the nonvariational character of dynamics. PMID- 14524944 TI - Optical second-harmonic generation enhanced by a twist defect in ferroelectric liquid crystals. AB - Second-harmonic generation (SHG) spectra were numerically calculated in ferroelectric liquid crystals with a twist defect. It is shown that SHG is enhanced when the SHG wavelength is close to the defect mode. The spectral width of the enhanced peak becomes sharper with increasing the sample thickness at the same rate for the width of the defect mode peak. The SHG peak intensity increases with about seventh power of the sample thickness. PMID- 14524945 TI - Balancing at the border of instability. AB - Some biological systems operate at the critical point between stability and instability, and this requires a fine tuning of parameters. We bring together two examples from the literature that illustrate this: neural integration in the nervous system and hair cell oscillations in the auditory system. In both examples the question arises as to how the required fine tuning may be achieved and maintained in a robust and reliable way. We study this question using tools from nonlinear and adaptive control theory. We illustrate our approach on a simple model which captures some of the essential features of neural integration. As a result, we propose a large class of feedback adaptation rules that may be responsible for the experimentally observed robustness of neural integration. We mention extensions of our approach to the case of hair cell oscillations in the ear. PMID- 14524946 TI - Antispirals in an artificial tissue of oscillatory cells. AB - In a tissue of oscillatory cells the active intracellular medium is surrounded by a membrane and the cells are separated by inactive extracellular medium. The synchronization properties of a system of such coupled oscillatory cells have been emulated using the light-sensitive Belousov-Zhabotinsky reaction. Experimental results for four coupled cells are confirmed by numerical simulations. We have furthermore demonstrated the existence of antispirals and antipacemaker waves with inward propagating waves in larger cell assemblies of this type. Such dynamical structures are extremely rare in homogeneous chemical systems where the generic behavior is normal spirals and target patterns with outward-moving waves. PMID- 14524947 TI - Phase diffusion and random walk interpretation of electromagnetic scattering. AB - The relaxation behavior of phase observables for different particle diffusion models is found to establish a ground for radioscience interpretations of coherent backscatter spectra. The characteristic function for a random walk process at twice the incident radiation wave number is associated with the complex amplitude of the scattered field from a medium containing refractive index fluctuations. The phase relaxation function can be connected to the evolution of the characteristic function and may describe the average regression of the scattered field from a spontaneous fluctuation undergoing turbulent mixing. This connection holds when we assume that the stochastic description of particle movements based on a diffusion model is valid. The phase relaxation function, when identified as the generalized susceptibility function of the fluctuation dissipation theorem, is related to the spectral density of the scattered field from steady-state fluctuations. PMID- 14524948 TI - Shattering transitions in collision-induced fragmentation. AB - We investigate the kinetics of nonlinear collision-induced fragmentation. We obtain the fragment mass distribution analytically by utilizing its traveling wave behavior. The system undergoes a shattering transition in which a finite fraction of the mass is lost to infinitesimal fragments (dust). The nature of the shattering transition depends on the fragmentation process. When the larger of the two colliding fragments splits, the transition is discontinuous and the entire mass is transformed into dust at the transition point. When the smaller fragment splits, the transition is continuous with the dust gaining mass steadily on the account of the fragments. At the transition point, the fragment mass distribution diverges algebraically for small masses, c(m) approximately m( alpha), with alpha=1.201 91 em leader. PMID- 14524949 TI - Particle swarms in gases: the velocity-average evolution equations from Newton's law. AB - The evolution equation for a generic average quantity relevant to a swarm of particles homogeneously dispersed in a uniform gas, is obtained directly from the Newton's law, without having recourse to the (intermediary) Boltzmann equation. The procedure makes use of appropriate averages of the term resulting from the impulsive force (due to collisions) in the Newton's law. When the background gas is assumed to be in thermal equilibrium, the obtained evolution equation is shown to agree with the corresponding one following from the Boltzmann equation. But the new procedure also allows to treat physical situations in which the Boltzmann equation is not valid, as it happens when some correlation exists (or is assumed) between the velocities of swarm and gas particles. PMID- 14524950 TI - Theoretical description of phase coexistence in model C60. AB - We have investigated the phase diagram of a pair interaction model of C60 fullerene [L. A. Girifalco, J. Phys. Chem. 96, 858 (1992)], in the framework provided by two integral equation theories of the liquid state, namely, the modified hypernetted chain (MHNC) implemented under a global thermodynamic consistency constraint, and the self-consistent Ornstein-Zernike approximation (SCOZA), and by a perturbation theory (PT) with various degrees of refinement, for the free energy of the solid phase. We present an extended assessment of such theories as set against a recent Monte Carlo study of the same model [D. Costa, G. Pellicane, C. Caccamo, and M. C. Abramo, J. Chem. Phys. 118, 304 (2003)]. We have compared the theoretical predictions with the corresponding simulation results for several thermodynamic properties such as the free energy, the pressure, and the internal energy. Then we have determined the phase diagram of the model, by using either the SCOZA, the MHNC, or the PT predictions for one of the coexisting phases, and the simulation data for the other phase, in order to separately ascertain the accuracy of each theory. It turns out that the overall appearance of the phase portrait is reproduced fairly well by all theories, with remarkable accuracy as for the melting line and the solid-vapor equilibrium. All theories show a more or less pronounced discrepancy with the simulated fluid solid coexistence pressure, above the triple point. The MHNC and SCOZA results for the liquid-vapor coexistence, as well as for the corresponding critical points, are quite accurate; the SCOZA tends to underestimate the density corresponding to the freezing line. All results are discussed in terms of the basic assumptions underlying each theory. We have then selected the MHNC for the fluid and the first-order PT for the solid phase, as the most accurate tools to investigate the phase behavior of the model in terms of purely theoretical approaches. It emerges that the use of different procedures to characterize the fluid and the solid phases provides a semiquantitative reproduction of the thermodynamic properties of the C60 model at issue. The overall results appear as a robust benchmark for further theoretical investigations on higher order C(n>60) fullerenes, as well as on other fullerene-related materials, whose description can be based on a modelization similar to that adopted in this work. PMID- 14524951 TI - Dynamic heterogeneities in the out-of-equilibrium dynamics of simple spherical spin models. AB - The response of spherical two-spin interaction models, the spherical ferromagnet (s-FM) and the spherical Sherrington-Kirkpatrick (s-SK) model, is calculated for the protocol of the so-called nonresonant hole burning (NHB) experiment for temperatures below the respective critical temperatures. It is shown that it is possible to select dynamic features in the out-of-equilibrium dynamics of both models, one of the hallmarks of dynamic heterogeneities. The behavior of the s-SK model and the s-FM model in three dimensions is very similar, showing dynamic heterogeneities in the long-time behavior, i.e., in the aging regime. The appearance of dynamic heterogeneities in the s-SK model explicitly demonstrates that these are not necessarily related to spatial heterogeneities. For the s-FM model, it is shown that the nature of the dynamic heterogeneities changes as a function of dimensionality. With the increasing dimension, the frequency selectivity of the NHB diminishes and the dynamics in the mean-field limit of the s-FM model becomes homogeneous. PMID- 14524952 TI - Hard sphere fluids at surfaces of porous media. AB - An adsorbate fluid of hard spheres is brought into contact with a semi-infinite porous matrix modeled by immobilized configurations of freely overlapping spheres with a sharp kink one-body density distribution. Comparison of results from a recent density-functional approach to those of our computer simulations yields good agreement for the adsorbate density profile across the matrix surface. We show how the matrix can be replaced by a fictitious external potential that only depends on the distance from the interface, and that leads to the same adsorbate density profile. This potential is found to be a smooth function of distance, due to the geometry of the matrix particles. For high matrix densities, the porous medium becomes practically impenetrable, and its surface behaves like a rough hard wall whose roughness decreases with increasing matrix density. PMID- 14524953 TI - Two-dimensional diffusion in the presence of topological disorder. AB - How topological defects affect the dynamics of particles hopping between lattice sites of a distorted, two-dimensional crystal is addressed. Perturbation theory and numerical simulations show that weak, short-ranged topological disorder leads to a finite reduction of the diffusion coefficient. Renormalization group theory and numerical simulations suggest that longer-ranged disorder, such as that from randomly placed dislocations or random disclinations with no net disclinicity, leads to subdiffusion at long times. PMID- 14524954 TI - Fluctuating nematic elastomer membranes. AB - We study the flat phase of nematic elastomer membranes with rotational symmetry spontaneously broken by an in-plane nematic order. Such a state is characterized by a vanishing elastic modulus for simple shear and soft transverse phonons. At harmonic level, the in-plane orientational (nematic) order is stable to thermal fluctuations that lead to short-range in-plane translational (phonon) correlations. To treat thermal fluctuations and relevant elastic nonlinearities, we introduce two generalizations of two-dimensional membranes in a three dimensional space to arbitrary D-dimensional membranes embedded in a d dimensional space and analyze their anomalous elasticities in an expansion about D=4. We find a stable fixed point that controls long-scale properties of nematic elastomer membranes. It is characterized by singular in-plane elastic moduli that vanish as a power law eta(lambda)=4-D of a relevant inverse length scale (e.g., wave vector) and a finite bending rigidity. Our predictions are asymptotically exact near four dimensions. PMID- 14524955 TI - Vortex dynamics in a coarsening two-dimensional XY model. AB - The vortex velocity distribution function for a two-dimensional coarsening nonconserved O(2) time-dependent Ginzburg-Landau model is determined numerically and compared to theoretical predictions. In agreement with these predictions the distribution function scales with the average vortex speed which is inversely proportional to t(x), where t is the time after the quench and x is near 1/2. We find the entire curve, including a large speed algebraic tail, in good agreement with the theory. PMID- 14524956 TI - Determination of multicanonical weight based on a stochastic model of sampling dynamics. AB - Based on the stochastic interpretation of the sampling process modeled by a Langevin equation, we present an effective iteration scheme to determine the weight in multicanonical molecular dynamics. Our method enables an automatic determination of the weight producing a uniform energy sampling via an iterative cancellation of the deterministic force in a Langevin equation. The deterministic force has been calculated from the energy trajectory by identifying the moments of the transition probability of a Fokker-Planck equation associated with a Langevin equation. The intimate relationship between the sampling process and the stochastic dynamics has been verified by applying the iteration scheme to a helix coil transition of the 8-polyalanine system in a gas phase. PMID- 14524957 TI - Superoperator representation of nonlinear response: unifying quantum field and mode coupling theories. AB - Computing response functions by following the time evolution of superoperators in Liouville space (whose vectors are ordinary Hilbert space operators) offers an attractive alternative to the diagrammatic perturbative expansion of many-body equilibrium and nonequilibrium Green's functions. The bookkeeping of time ordering is naturally maintained in real (physical) time, allowing the formulation of Wick's theorem for superoperators, giving a factorization of higher order response functions in terms of two fundamental Green's functions. Backward propagations and analytic continuations using artificial times (Keldysh loops and Matsubara contours) are avoided. A generating functional for nonlinear response functions unifies quantum field theory and the classical mode coupling formalism of nonlinear hydrodynamics and may be used for semiclassical expansions. Classical response functions are obtained without the explicit computation of stability matrices. PMID- 14524958 TI - Vicious walks with a wall, noncolliding meanders, and chiral and Bogoliubov-de Gennes random matrices. AB - Spatially and temporally inhomogeneous evolution of one-dimensional vicious walkers with wall restriction is studied. We show that its continuum version is equivalent with a noncolliding system of stochastic processes called Brownian meanders. Here the Brownian meander is a temporally inhomogeneous process introduced by Yor as a transform of the Bessel process that is the motion of radial coordinate of the three-dimensional Brownian motion represented in spherical coordinates. It is proved that the spatial distribution of vicious walkers with a wall at the origin can be described by the eigenvalue statistics of Gaussian ensembles of Bogoliubov-de Gennes Hamiltonians of the mean-field theory of superconductivity, which have a particle-hole symmetry. We report that a time evolution of the present stochastic process is fully characterized by the change of symmetry classes from type C to type CI in the nonstandard classes of random matrix theory of Altland and Zirnbauer. The relation between the noncolliding systems of the generalized meanders of Yor, which are associated with the even-dimensional Bessel processes, and the chiral random matrix theory is also clarified. PMID- 14524959 TI - Growth-based optimization algorithm for lattice heteropolymers. AB - An improved version of the pruned-enriched-Rosenbluth method (PERM) is proposed and tested on finding lowest energy states in simple models of lattice heteropolymers. It is found to outperform not only the previous version of PERM, but also all other fully blind general purpose stochastic algorithms which have been employed on this problem. In many cases, it found new lowest energy states missed in previous papers. Limitations are discussed. PMID- 14524960 TI - Structure of liquids composed of shifted dipole linear molecules. AB - Simulation results for liquids composed of linear molecules interacting through dispersion forces and off-center dipoles are presented. Remarkable differences are found on the vapor-liquid equilibrium respect to that of centered dipole molecules. Even more remarkable is the appearance of additional short-range liquid structure at relatively large dipoles and aspect ratios. The existence of dipole dimers is clearly established, and some suggestions allowing for the correspondence between a particular macroscopic phenomenology and a particular dispersion potential function are presented. PMID- 14524961 TI - Binary and multiparticle contributions to the velocity autocorrelation function. AB - A method for including the contribution of many-body correlation effects to the microscopically obtained results of the two-body contribution to the velocity autocorrelation has been proposed. A significant improvement over the results obtained through only binary contribution has been found, as can be judged by comparing the results for force and velocity autocorrelation functions of Lennard Jones fluids with that of molecular dynamic simulations. The agreement of results of self-diffusion coefficient is also quite good with simulation data over a wide range of densities and temperatures. PMID- 14524962 TI - System size dependence of the diffusion coefficient in a simple liquid. AB - An equation to estimate the system size dependence of the self-diffusion coefficient of a tagged particle moving in a simple fluid is given using linear response theory and linearized hydrodynamics. Estimates made by the equation are compared with the results of the molecular dynamics simulation for a hard-sphere fluid at two densities, rhosigma(3) approximately 0.88 and 0.47, where sigma is the hard-sphere diameter. Good agreement between theory and simulation is obtained at the higher density. At the lower density, the agreement becomes poorer, but it is improved by taking into account the diffusion effect of the tagged particle. The equation gives the same diffusion coefficient for the infinite system as that obtained by taking into account the long-time tail contribution of the velocity autocorrelation function [B. J. Alder, D. M. Gass, and T. E. Wainwright, J. Chem. Phys. 53, 3813 (1970)]. When the tagged particle has a larger mass than the fluid particles, the equation presented here gives the better estimates. It is confirmed by the molecular dynamics calculation. PMID- 14524963 TI - Partially fluidized shear granular flows: continuum theory and molecular dynamics simulations. AB - The continuum theory of partially fluidized shear granular flows is tested and calibrated using two-dimensional soft particle molecular dynamics simulations. The theory is based on the relaxational dynamics of the order parameter that describes the transition between static and flowing regimes of granular material. We define the order parameter as a fraction of static contacts among all contacts between particles. We also propose and verify by direct simulations the constitutive relation based on the splitting of the shear stress tensor into a"fluid part" proportional to the strain rate tensor, and a remaining "solid part." The ratio of these two parts is a function of the order parameter. The rheology of the fluid component agrees well with the kinetic theory of granular fluids even in the dense regime. Based on the hysteretic bifurcation diagram for a thin shear granular layer obtained in simulations, we construct the "free energy" for the order parameter. The theory calibrated using numerical experiments with the thin granular layer is applied to the surface-driven stationary two-dimensional granular flows in a thick granular layer under gravity. PMID- 14524964 TI - Fractional diffusion model for force distribution in static granular media. AB - We present the results of a numerical and an experimental investigation of the probability distribution of normal contact forces in static packs of particles with two different hardnesses. Force distributions are computed and compared with existing models and experimental data. It is found that the probability distribution function of normal contact forces P(f) is well described by a semiempirical model derived from a fractional diffusion equation. This model reproduces most of the features common to force distributions observed in experimental and numerical studies including the finite value for P(f) as the forces tend to zero. The results indicate that the fractional model fits well both the numerical and experimental data over a wide range of particle deformations in contrast to the existing models. These results provide an insight into the physics of granular media and complement previous findings. PMID- 14524965 TI - Dynamics of two granules. AB - We study the dynamics of two particles that interact only when in contact. In this sense, although not in every particular, the interactions mimic those in granular materials. The detailed solution of the dynamics allows an analysis of the backscattering behavior of the first particle and of the energy dissipation in the system as a function of various parameters. PMID- 14524966 TI - Mass and size effects in three-dimensional vibrofluidized granular mixtures. AB - We examine the steady state properties of binary systems of driven inelastic hard spheres. The spheres, which move under the influence of gravity, are contained in a vertical cylinder with a vibrating base. We computed the trajectories of the spheres using an event-driven molecular dynamics algorithm. In the first part of the study, we chose simulation parameters that match those of experiments published by Wildman and Parker. Various properties computed from the simulation including the density profile, granular temperature, and circulation pattern are in good qualitative agreement with the experiments. We then studied the effect of varying the mass ratio and the size ratio independently while holding the other parameters constant. The mass and size ratio are shown to affect the distribution of the energy. The changes in the energy distributions affect the packing fraction and temperature of each component. The temperature of the heavier component has a nonlinear dependence on the mass of the lighter component, while the temperature of the lighter component is approximately proportional to its mass. The temperature of both components is inversely dependent on the size of the smaller component. PMID- 14524967 TI - Brownian motion in confinement. AB - This research modifies an earlier approach based on the single-wall reflection method to predict the perpendicular and parallel diffusion coefficients of a Brownian sphere in confinement. The modified version provides predictions that match experimental data reported in the literature more accurately than those provided by other available models, including the linear superposition approximation, the coherent superposition approximation, and Oseen's equation. PMID- 14524968 TI - Mechanics of semiflexible chains formed by poly(ethylene glycol)-linked paramagnetic particles. AB - Magnetorheological particles, permanently linked into chains, provide a magnetically actuated means to manipulate microscopic fluid flow. Paramagnetic colloidal particles form reversible chains by acquiring dipole moments in the presence of an external magnetic field. By chemically connecting paramagnetic colloidal particles, flexible magnetoresponsive chains can be created. We link the paramagnetic microspheres using streptavidin-biotin binding. Streptavidin coated microspheres are placed in a flow cell and a magnetic field is applied, causing the particles to form chains. Then a solution of polymeric linkers of bis biotin-poly(ethylene glycol) molecules is added in the presence of the field. These linked chains remain responsive to a magnetic field; however, in the absence of an external magnetic field these chains bend and flex due to thermal motion. The chain flexibility is determined by the length of the spacer molecule between particles and is quantified by the flexural rigidity or bending stiffness. To understand the mechanical properties of the chains, we use a variety of optical trapping experiments to measure the flexural rigidity. Increasing the length of the poly(ethylene glycol) chain in the linker increases the flexibility of the chains. PMID- 14524969 TI - Transport properties of incipient gels. AB - We investigate the behavior of the shear viscosity eta(p) and the mass-dependent diffusion coefficient D(m,p) in the context of a simple model that, as the cross link density p is increased, undergoes a continuous transition from a fluid to a gel. The shear viscosity diverges at the gel point according to eta(p) approximately (p(c)-p)(-s) with s approximately 0.65. The diffusion constant shows a remarkable dependence on the mass of the clusters: D(m,p) approximately m(-0.69), not only at p(c) but well into the liquid phase. We also find that the Stokes-Einstein relation Deta proportional, variant k(B)T breaks down already quite far from the gel point. PMID- 14524970 TI - Testing the Derjaguin approximation for colloidal mixtures of spheres and disks. AB - The depletion potential between two large hard spheres due to the presence of hard disks has been derived up to first order in the number density of disks by Piech and Walz [J. Colloid Interface Sci. 232, 86 (2000)] using the Derjaguin approximation. Using the generalized Gibbs equation, we compare this depletion potential to the exact solution up to first order in density. The Derjaguin approximation turns out to be surprisingly accurate; for aspect ratios smaller than 0.25 the error is less than 1%. PMID- 14524971 TI - Tuning the interactions of a magnetic colloidal suspension. AB - We present a versatile experimental system of magnetic charged nanospheres dispersed in water that belongs to both dipolar and electrostatic systems. In this system, the interactions can be continuously tuned by varying the ionic strength I. At low I, the potential is a strongly repulsive Yukawa potential that leads to a phase diagram similar to the one of repulsive spheres (fluid and solid phases). At high I, the potential is a globally attractive Lennard-Jones potential that leads to a phase diagram similar to the one of atomic systems (gas, liquid, fluid, and solid phases). PMID- 14524972 TI - Hydration-dependent internal dynamics of reverse micelles: a quasielastic neutron scattering study. AB - We studied the overall atomic mobility of sodium bis-(2-ethylhexyl) sulfosuccinate (AOT) reverse micelles in deuterated cyclohexane (C6D12) as a function of the molar ratio W=[D2O]/[AOT] with an incoherent quasielastic neutron scattering experiment at high energy resolution. For the almost anhydrous sample, the quasielastic broadening can be entirely attributed to the reverse micelle global motion, by considering explicitly both the rotational and the translational terms. As W increases above a threshold value W approximately 1 a wide quasielastic signal appears, which has been interpreted as the onset of a hydration-dependent intrinsic micelle dynamics. Such a contribution, which involves the AOT monomer hydrogen atoms, has a characteristic time of 0.2 ns. This result has been compared with previous dielectric measurements, which detected a relaxation process of the AOT fully hydrated head groups with the same characteristic time. The internal macromolecule mobility evaluated as a function of W numerically correlates with that of the mobile head groups, calculated by dielectric measurements. These findings suggest that both the hydrophobic and hydrophilic moieties dynamics is activated by the progressive hydration of the reverse micelle. PMID- 14524973 TI - Phase diagrams of hard-core repulsive Yukawa particles. AB - We determine the phase behavior of hard spheres interacting with repulsive Yukawa (screened Coulomb) interaction using computer simulations. We study the effect of the hard-core diameter on the phase behavior of repulsive Yukawa particles by comparing our phase diagrams with that of repulsive point Yukawa particles. We show that for sufficiently high contact values of the pair potential (betaepsilon=20, 39, 81, and higher), the fluid-face-centered-cubic (fcc) solid, at high screening, the fluid-body-centered-cubic (bcc) solid and the bcc-fcc coexistence for packing fractions eta less, similar 0.5 are well described by the phase boundaries of point Yukawa particles, by employing a mapping of the point Yukawa system onto a hard-core repulsive Yukawa system. While the bcc-fcc coexistence is well described by the point Yukawa limit for eta<0.5, we find a deviation at higher eta as the hard-core repulsion favors the fcc solid for eta>/=0.5, independent of the screening. Consequently, a second triple point appears in the phase diagram in the weak screening regime. In addition, we find that all the phase coexistence regions in our phase diagrams for hard-core repulsive Yukawa system are very narrow, i.e., a small density jump in the coexisting phases. PMID- 14524974 TI - Granular dynamics of density profiles in a suspension interface. AB - The temporal evolution of the density profiles at the interface of a Rayleigh Taylor instability in a sedimenting suspension is experimentally investigated. It is found that the sand-glycerin density gradients within the interface change with time, and that the evolution of the gradients differs significantly, depending on the location: The density profiles become steeper in the regions where the suspension flows downwards, whereas the profiles become flatter in the regions where the fluid flows upwards. This observation shows that there is a motion of the sand grains relative to the carrier fluid and hence reveals the prevailing granular dynamics in the suspension interface. It shows a behavior of the suspension which is different from the behavior of a homogeneous Newtonian one-component fluid whose density profiles do not change in time. Another interesting result is that the slopes of the profiles differ already at the very beginning (t=0) of the evolution of the instability, indicating that the suspension might have self-structured prior to instability onset. PMID- 14524975 TI - Calorimetric study of magnetic fluids under a magnetic field. AB - In our past study, it was found that the strong magneto-optical effect of a magnetic fluid (MF) under a magnetic field is due to the second-order phase transition from colloidal particles' monodispersed phase to the particles' anisotropically agglomerated microclusters phase. These results, however, contradicted the Landau criterion. In the present study, in order to clarify whether the transition is of first order or second order, we performed differential scanning calorimetry (DSC) experiments on three different MFs to look for the existence of a phase transition latent heat. The DSC measurement was performed in the temperature range 22-150 degrees C with zero magnetic field and under approximately 10 kA/m magnetic field, respectively. No phase transition heat was observed within an experimental error of 0.03 kJ/kg for all the samples whether or not the field was applied. PMID- 14524976 TI - Glass transition of hard sphere systems: molecular dynamics and density functional theory. AB - The glass transition of a hard sphere system is investigated within the framework of the density functional theory (DFT). Molecular dynamics (MD) simulations are performed to study the dynamical behavior of the system on the one hand and to provide the data to produce the density field for the DFT on the other hand. Energy landscape analysis based on the DFT shows that there appears a metastable (local) free energy minimum representing an amorphous state as the density is increased. This state turns out to become stable, compared with the uniform liquid, at some density around which we also observe a sharp slowing down of the alpha relaxation in the MD simulations. PMID- 14524977 TI - Changes in dynamic crossover with temperature and pressure in glass-forming diethyl phthalate. AB - Dielectric relaxation measurements have been used to study the crossover in dynamics with temperature and pressure, onset of breakdown of the Debye-Stokes Einstein law, and the relation between the alpha and the beta relaxations in diethyl phthalate. The measurements made over 10 decades in frequency and a broad range of temperature and pressure enable the dc conductivity and the alpha- and the beta-relaxations to be studied altogether. The isobaric data show that the alpha-relaxation time tau(alpha) has temperature dependence that crosses over from one Vogel-Fulcher-Tammann-Hesse form to another at T(B) approximately 227 K and tau(alpha) approximately 10(-2) s. The dc conductivity sigma exhibits similar crossover at the same T(B). At temperatures above T(B), tau(alpha) and sigma have the same temperature dependence, but below T(B) they become different and the Debye-Stokes-Einstein law breaks down. The breadth of the alpha relaxation is nearly constant for TT(B). The time dependence of tau(beta) is Arrhenius, which when extrapolated to higher temperatures intersects tau(alpha) at T(beta) nearly coincident with T(B). Isothermal measurements at various applied pressures when compared with isobaric data show that the shape of the alpha-relaxation depends only on tau(alpha), and not on the T and P combinations. At a constant temperature, while tau(alpha) increases rapidly with pressure, the beta-relaxation time tau(beta) is insensitive to applied pressure. This behavior is exactly the same as found in 1,1(')-bis (p-methoxyphenyl) cyclohexane. The findings are discussed in the framework of the coupling model. PMID- 14524978 TI - Nanostructures of polyelectrolyte gel-surfactant complexes in uniaxially stretched networks. AB - Nanostructures of poly(acrylate) gel and dodecylpyridinium complexes equlibrated with the NaCl aqueous solution (from 5 to 100mM) and their time evolution after stretching uniaxially were investigated by means of time-resolved small-angle x ray scattering. The scattering profile revealed the existence of the cubic nanostructure belonging to Pm3n space group in the gel before and long after the stretch. Each of the three intensive peaks was found to be resolved into two, which suggested the existence of two cubic structures with the slightly different lattice spacings. On the other hand, the splits of scattering peaks were not observed for the linear poly(acrylate) and dodecylpyridinium complexes. This indicates that the existence of the cross-linked chain is concerned in the formation of the double structure in the complex system. A series of time resolved experiments demonstrated that the peaks corresponding to the longer lattice constant disappeared once just after stretching and regenerated to grow up, whereas the peaks corresponding to the shorter lattice constant were continuously observed. The growing rates of the peaks increased with the NaCl concentration. It was also found that a two-dimensional scattering pattern changed from the Debye-Scherrer ring type into the Laue spot type with stretching at the lower NaCl concentrations. This indicates that the single-crystal-like domains align in the stretched network due to the strong electrostatic interaction between the dodecylpyridinium cation and poly(acrylate) anion. PMID- 14524979 TI - Self-consistent molecular field theory for packing in classical liquids. AB - Building on a quasichemical formulation of solution theory, this paper proposes a self-consistent molecular field theory for packing problems in classical liquids, and tests the theoretical predictions for the excess chemical potential of the hard sphere fluid. Results are given for the self-consistent molecular fields obtained, and for the probabilities of occupancy of a molecular observation volume. For this system, the excess chemical potential predicted is nearly as accurate as the most accurate prior theories, specifically the scaled particle (Percus-Yevick compressibility) theory. A compact formula is obtained to throw light on the variation of the chemical potential with the radius of a distinguished hard sphere solute in general solvents. It is argued that the present approach is particularly simple, permits a natural description of possibilities for multiphasic behavior of the solution, and should provide a basis for a molecular-scale description of more complex solutions. PMID- 14524980 TI - Dynamical scaling of surface growth in simple lattice models. AB - We present extensive simulations of the atomistic Edwards-Wilkinson (EW) and Restricted Edwards-Wilkinson (REW) models in 2+1 dimensions. Dynamic finite-size scaling analyses of the interfacial width and structure factor provide the estimates for the dynamic exponent z=1.65+/-0.05 for the EW model and z=2.0+/-0.1 for the REW model. The stochastic contribution to the interface velocity U due to the deposition and diffusion of particles is characterized for both the models using a blocking procedure. For the EW model the time-displaced temporal correlations in U show nonexponential decay, while the temporal correlations decay exponentially for the REW model. Dynamical scaling of the temporal correlation function for the EW model yields a value of z, which is consistent with the estimate obtained from finite-size scaling of the interfacial width and structure factor. PMID- 14524981 TI - Transitions between smooth and complex stick-slip sliding of surfaces. AB - Shear measurements were performed on mica surfaces with molecularly thin films of squalane (C30H62) confined between them. Squalane is a branched hydrocarbon liquid that can be in the liquid, glassy, or liquid-crystalline state under confinement. The friction forces, especially the transitions between smooth and intermittent (e.g., stick-slip) sliding, were measured over a wider range of applied loads (pressures), sliding velocities (shear rates), and temperatures than in previous studies. The results reveal that, depending on the conditions, qualitatively different behavior can arise in the same system. These include both abrupt and continuous transitions, both upper and lower critical transition temperatures, short and very long transient effects, and chaotic, sawtooth, or sinusoidal stick-slip that can slowly decay with time or distance sheared. The differences between these branched and simpler, e.g., spherical, unbranched molecules are compared, as well as with unlubricated (dry) surfaces and macroscopic (geological) systems. PMID- 14524982 TI - Pattern formation in crystal growth under parabolic shear flow. AB - Morphological instability of the solid-liquid interface occurring in a crystal growing from an undercooled thin liquid bounded on one side by a free surface and flowing down inclined plane, is investigated by a linear stability analysis under shear flow. It is found that restoring forces due to gravity and surface tension is an important factor for stabilization of the solid-liquid interface on long length scales. This is a stabilizing effect different from the Gibbs-Thomson effect. A particular long wavelength mode of about 1 cm of wavy pattern, observed on the surface of icicles covered with a thin layer of flowing water is obtained from the dispersion relation, including the effect of flow and restoring forces. PMID- 14524983 TI - Phase field models for step flow. AB - The relation between phase field and discontinuous models for crystal steps is analyzed. Different formulations of the kinetic boundary conditions of the discontinuous model are first presented. We show that (i) step transparency, usually interpreted as the possibility for adatoms to jump through steps, may be seen as a modification of the equilibrium concentration engendered by step motion. (ii) The interface definition (i.e., the position of the dividing line) intervenes in the expression of the kinetic coefficients only in the case of fast attachment kinetics. (iii) We also identify the thermodynamically consistent reference state for kinetic boundary conditions. Asymptotic expansions of the phase field models in the limit where the interface width is small, lead to various discontinuous models. (1) A phase field model with one global concentration field and variable mobility is shown to lead to a discontinuous model with fast step kinetics. (2) A phase field model with one concentration field per terrace allows one to recover arbitrary step kinetics (i.e., arbitrarily strong Ehrlich-Schwoebel effect and step transparency). Quantitative agreement is found, in both the linear and nonlinear regimes, between the numerical solution of the phase field models and the analytical solution of the discontinuous model. PMID- 14524984 TI - Nucleation preexponential in dynamic Ising models at moderately strong fields. AB - A dynamic Ising model on a two-dimensional square lattice with nearest neighbor interactions is considered in the metastable region at low temperatures. A large number of low-energy cluster configurations is identified, and for those configurations a system of kinetic equations is written. Solution is obtained using symbolic computational approaches. This allows one to identify the full expression for the nucleation rate, including the preexponential. The treatment generalizes the earlier study of a different, lattice-gas spin-flip dynamics [V. A. Shneidman and G. M. Nita, Phys. Rev. Lett. 89, 025701 (2002)], for the cases of Glauber and Metropolis dynamics and for a broader region of fields. In addition, connection with the lowest-energy nucleation paths (which can be studied analytically, without computer assistance) is examined. PMID- 14524985 TI - Crossover effects in the wetting of adsorbed films in linear wedges. AB - We have measured the growth of liquid Ar adsorbed on arrays of linear wedges structured in different ways. In the most regular patterns, a clear crossover from a planarlike to a geometry-dependent growth behavior is observed. This crossover is found to depend on the characteristic wedge size and its position, in the case of a regular pattern, agrees well with theoretical predictions. Near liquid-vapor bulk coexistence, the film mass is observed to diverge as a power law of the chemical potential difference from saturation with an exponent in very good agreement with the value of -2 expected for a linear wedge. This exponent is not affected by the opening angles of the wedges. The form of the next-to-leading order singular term in the asymptotic divergence of the mass has also been investigated. The experimentally determined value of the exponent is consistent with the expected theoretical result of -4/3. PMID- 14524986 TI - Three-dimensional nature of ion transport in thin-layer electrodeposition. AB - A generalized three-dimensional model for ion transport in electrodeposition is introduced. Ion transport is mainly governed by diffusion, migration, and convection. When convection prevails, in particular, in the limiting case of gravity-driven convection, the model predicts concentration shells and convection rolls and their interaction mode with a deposit tip: shell and roll bend and surround the tip forming a three-dimensional envelope tube squeezed at the deposit tip. In the limiting case of electrically driven convection, a vortex ring and an electric spherical drop crowning the deposit tip are predicted. When gravity and electric convection are both relevant, the interaction of ramified deposits, vortex tubes and rings, and electric spherical drops, leading to complex helicoidal flow, is predicted. Many of these predictions are experimentally observed, suggesting that ion transport underlying dendrite growth is remarkably well captured by our model. PMID- 14524987 TI - Shape and director-field transformation of tactoids. AB - Tactoids are droplets of a nematic phase that under suitable conditions form in dispersions of elongated colloidal particles. We theoretically study the shape and the director-field configuration of such droplets for the case where a planar anchoring of the director field to the interface is favored. A minimum of four regimes can be identified in which the droplets have a different structure. Large droplets tend to be nearly spherical with a director field that is bipolar if the surface tension is strongly anisotropic and homogeneous if this is not so. Small droplets can become very elongated and spindlelike if the surface tension is sufficiently anisotropic. Depending on the anchoring strength, the director field is then either homogeneous or bipolar. We find that the more elongated the tactoid, the more strongly it resists the crossing over from a homogeneous to a bipolar structure. This should have implications for the nucleation rate of the nematic phase. Our calculations qualitatively describe the size dependence of the aspect ratio of tactoids found in recent experiments. PMID- 14524988 TI - Optical rotatory power, biaxiality, and models of chiral tilted smectic phases. AB - Among the chiral tilted smectics, the stable existence has been confirmed in numerous investigations of SmC(*)(A), (antiferroelectric smectic-C(A)) SmC(*)(F11) (SmC(*)(gamma)), SmC(*)(F12) (antiferroelectric, AF) and SmC* phases. The structures of the ferrielectric SmC(*)(F11) and SmC(*)(F12) phases suggested by different models are essentially different although all the models use the three-layer and four-layer periodicity for them. The structures of the phases were investigated using the optical rotatory power (ORP) measurements technique. The ORP was simulated using Berreman's 4 x 4-matrix method. The compound under investigation (S)-1-methylheptyl 4-(4(')-n-undecyloxy-biphenyl-4-yl-carbonyloxy) [acronym (S)-11OF1M7] clearly provides SmC(*)(F11) and SmC(*)(F12) phases, the temperature range for the existence of these phases is about 5 degrees C each. This had not been achieved for the earlier investigated antiferroelectric liquid crystal (AFLC) samples. The results obtained confirm that the unit cell of the molecular structure of these subphases is highly biaxial. Due to the biaxiality the texture of the homeotropic cell under a polarizing microscope appears nonuniform. This requires a special approach to the measurements and a simulation of the ORP, which is discussed in detail. A technique has been designed where the transmitted intensity through a polarizing microscope is measured as a function of the angle of polarization of the incident light. From the observed output, which is a biased sine wave, the ORP is being determined. In the same scan, the wavelength of light is also being automatically altered. Comparing the simulated and measured data, we can conclude that in the SmC(*)(F12) phase the distortion angle of the directors in the Ising model is lower than 10 degrees. Using the Ising model, the pitch in SmC(*)(F11) has been determined and this is found to have a strong temperature dependence. PMID- 14524989 TI - Spin-lattice dispersion in nematic and smectic-A mesophases in the presence of ultrasonic waves: a theoretical approach. AB - We present a theoretical study of the Larmor frequency dependence of the nuclear spin-lattice relaxation caused by order director fluctuations for both nematic and smectic-A mesophases. The analysis is focused on the case where the molecular system is subjected to sonication during the relaxation process. The departure from the nonsonicated case is discussed for various values of the involved parameters. Two different approaches are discussed for the smectic case. PMID- 14524990 TI - Pseudo-Casimir force in chiral smectic liquid crystals. AB - We present a theoretical study of the pseudo-Casimir force in two chiral smectic systems: a homeotropic cell and a free-standing film. We consider the interaction induced by the fluctuations of orientational order. We demonstrate how the character of the force depends on the type of fluctuation modes and on boundary conditions. We focus on the temperature dependence of the force, which is marked by the vicinity of the smectic-A*-->smectic-C* phase transition. We find that at this transition the force diverges if the system is frustrated; otherwise it remains finite. We expose the analogy between the force in these smectic systems and in previously studied nematic systems, thus demonstrating the universality of the pseudo-Casimir interaction. PMID- 14524991 TI - Calorimetric study of octylcyanobiphenyl liquid crystal confined to a controlled pore glass. AB - We present a calorimetric study of the phase behavior of octylcyanobiphenyl (8CB) liquid crystal confined to a controlled-pore glass (CPG). We used CPG matrices with characteristic void diameters ranging from 400 to 20 nm. In bulk we obtain weakly first-order isotropic to nematic (I-N) phase transition and nearly continuous character of the nematic to smectic-A (N-SmA) phase transition. In all CPG matrices the I-N transition remains weakly first order, while the N-SmA one becomes progressively suppressed with decreasing CPG pore radius. With decreased pore diameters both phase transition temperatures monotonously decrease following similar trends, but increasing the stability range of the N phase. The heat capacity response at the weakly first order I-N and continuous N-SmA phase transitions gradually approaches the tricritical-like and three-dimensional XY behavior, respectively. The main observed features were explained using a bicomponent single pore type phenomenological model. PMID- 14524992 TI - Molecular simulation of chevrons in confined smectic liquid crystals. AB - Chevron structures adopted by confined smectic liquid crystals are investigated via molecular dynamics simulations of the Gay-Berne model. The chevrons are formed by quenching nematic films confined between aligning planar substrates whose easy axes have opposing azimuthal components. When the substrates are perfectly smooth, the chevron formed migrates rapidly towards one of the confining walls to yield a tilted layer structure. However, when substrate roughness is included, by introducing a small-amplitude modulation to the particle-substrate interaction well depth, a symmetric chevron is formed which remains stable over sufficiently long run times for detailed structural information, such as the relevant order parameters and director orientation, to be determined. For both smooth and rough boundaries, the smectic order parameter remains nonzero across the entire chevron, implying that layer identity is maintained across the chevron tip. Also, when the surface-stabilized chevron does eventually revert to a tilted layer structure, it does so via surface slippage, such that layer integrity is maintained throughout the chevron to tilted layer relaxation process. PMID- 14524993 TI - Order reconstruction in frustrated nematic twist cells. AB - Within the Landau-de Gennes theory of liquid crystals, we study the equilibrium configurations of a nematic cell with twist boundary conditions. Under the assumption that the order tensor Q be uniaxial on both bounding plates, we find three separate classes of solutions, one of which contains the absolute energy minimizer, a twistlike solution that exists for all values of the distance d between the plates. The solutions in the remaining two classes exist only if d exceeds a critical value d(c). One class consists of metastable, twistlike solutions, while the other consists of unstable, exchangelike solutions, where the eigenvalues of Q are exchanged across the cell. When d=d(c), the metastable solution relaxes back to the absolute energy minimizer, undergoing an order reconstruction somewhere within the cell. The critical distance d(c) equals, in general, a few biaxial coherence lengths. This scenario applies to all the values of the boundary twist but pi/2, which thus appears as a very special case, though it is the one more studied in the literature. In fact, when the directors prescribed on the two plates are at right angles, two symmetric twistlike solutions merge continuously into an exchangelike solution at the critical value of d where the latter becomes unstable. Our analysis shows how the classical bifurcation associated with this phenomenon is unfolded by perturbing the boundary conditions. PMID- 14524994 TI - Computer simulations of hard pear-shaped particles. AB - We report results obtained from Monte Carlo simulations investigating mesophase formation in two model systems of hard pear-shaped particles. The first model considered is a hard variant of the truncated Stone-expansion model previously shown to form nematic and smectic mesophases when embedded within a 12-6 Gay Berne-like potential [R. Berardi, M. Ricci, and C. Zannoni, ChemPhysChem 7, 443 (2001)]. When stripped of its attractive interactions, however, this system is found to lose its liquid crystalline phases. For particles of length to breadth ratio k=3, glassy behavior is seen at high pressures, whereas for k=5 several bi- layerlike domains are seen, with high intradomain order but little interdomain orientational correlation. For the second model, which uses a parametric shape parameter based on the generalized Gay-Berne formalism, results are presented for particles with elongation k=3, 4, and 5. Here, the systems with k=3 and 4 fail to display orientationally ordered phases, but the system with k=5 shows isotropic, nematic and, unusual for a hard-particle model, interdigitated smectic A2 phases. PMID- 14524995 TI - Mechanical phase diagram of shrinking cylindrical gels. AB - When polymer gels are subjected to an external stimulus such as temperature or solvent change, they undergo a phase transition often driving pattern formation. In this paper, we use an elastic model to investigate the linear stability of shrinking cylindrical NIPA gels. This model exhibits bubble and bamboo patterns. The wavelengths of these patterns and their phase diagram are in agreement with the experiment of Matsuo and Tanaka [Nature (London) 358, 482 (1992)]. PMID- 14524996 TI - Fracture behavior of Lennard-Jones glasses. AB - The fracture behavior of binary Lennard-Jones (LJ) glasses is studied by extensive molecular dynamics simulations. These LJ glasses represent a nonbond limit of polymer network glasses. We determine that the low strain behavior of the LJ and polymer glasses is similar. Two different LJ glasses are fractured under tensile strain without any preexisting crack. Void formation and resulting growth as strain increases is the mechanism through which the system fails. Void formation initiates at the yield strain of varepsilon (y) approximately 0.09, which is approximately the same strain at which the yielding behavior is first observed in cross-linked network models of polymer adhesives. The yield stress increased only by small amounts with increased strain rate and with increased system size (from N=30 000 atoms to 120 000 atoms). Within the ranges tested, the stress-strain behavior of these systems is independent of the temperature drop during quench and the initial molecular configuration. PMID- 14524997 TI - Structure factor of polymers interacting via a short range repulsive potential: application to hairy wormlike micelles. AB - We use the random phase approximation to compute the structure factor S(q) of a solution of chains interacting through a soft and short range repulsive potential V. Above a threshold polymer concentration, whose magnitude is essentially controlled by the range of the potential, S(q) exhibits a peak whose position depends on the concentration. We take advantage of the close analogy between polymers and wormlike micelles and apply our model, using a Gaussian function for V, to quantitatively analyze experimental small angle neutron scattering profiles of solutions of hairy wormlike micelles. These samples, which consist in surfactant self-assembled flexible cylinders decorated by amphiphilic copolymer, provide indeed an appropriate experimental model system to study the structure of sterically interacting polymer solutions. PMID- 14524998 TI - Static dynamics approach to relaxation modes and times for deformed polymers. AB - We discuss relaxation of conformational fluctuations around deformed polymer states. To this end, Brownian dynamics simulations of bead-spring models including a finite extensibility of the springs as well as excluded volume and hydrodynamic interactions between the beads have been performed. Complete spectra of relaxation times as well as corresponding relaxation modes are obtained from the simulation data by applying the static dynamics formalism, which rigorously describes the initial decay of correlations between the bead positions. As shown here, this procedure amounts to using a generalized Rouse-Zimm-like model which is governed by linear effective equations of motion having the same initial decay of correlations as the full nonlinear bead-spring model used in the simulations. In thermal equilibrium, the well-known scaling laws in the presence of excluded volume and hydrodynamic interactions between the beads are recovered, but, in addition, the static dynamics method also yields numeric values for the nonuniversal prefactors of the respective laws. The method is equally applicable to a broad range of problems, where the polymer is deformed by the action of flows or forces. Two examples of recent interest are considered: a tethered polymer pulled at its free end and one which is stretched by a uniform flow. It is shown that in both cases, the relaxation process is dominated by a finite extensibility of the springs. PMID- 14524999 TI - Effects of geometric confinement on the adhesive debonding of soft elastic solids. AB - The effect of increasing confinement on soft elastic gel layers has been investigated and a means of analyzing the behavior of such systems has been developed. A probe tack test was used to study the behavior of thin elastic layers during interfacial debonding from a cylindrical glass indenter. For this gel-indenter system, confinement is defined as the ratio of a(0), the radius of the indenter, to h, the thickness of the elastic layer. In order to investigate geometric effects, the adhesion energy of the gel was kept constant while the thickness and modulus of the gels were varied. A fracture mechanics approach, based on the compliance of the layer, has been employed in analyzing the experimental data. It is shown that a fracture mechanics analysis is appropriate for these systems, allowing quantitative results to be obtained, despite very irregular contacts. It has also been shown that the interfacial instabilities observed during debonding maximize the compliance of the elastic layer. Additionally, four dimensionless parameters that dictate the behavior of confined systems have been defined, offering a general guide to understanding and characterizing the mechanical behavior of thin elastic layers. PMID- 14525000 TI - Perturbation model to predict the effect of spatially varying absorptive inhomogeneities in diffusing media. AB - We develop a perturbation model to predict the effect of a spatially varying absorptive inhomogeneities in a diffusing slab. The model is based on a perturbation solution of diffusion equation derived for a refractive index mismatch between the scattering slab and the surrounding medium, through the use of the extrapolated boundary conditions. We show that the model allows to compute the time-dependent relative change in the transmitted signal resulting from the presence of the inclusion. We derive simplified expressions for the perturbed time-resolved transmittance that allows to implement an efficient fitting procedure for obtaining the optical properties of the absorptive inclusion. The accuracy of the predictions of the model was investigated through comparison with the results of the Finite Element Method to solve the time-dependent diffusion equation numerically. The procedure is used to obtain the absorption perturbation parameter of an absorptive inclusion characterized by spatially dependent Gaussian distribution of its absorption coefficient located at the midplane of a scattering slab. PMID- 14525001 TI - Theoretical predictions for spatial covariance of the electroencephalographic signal during the anesthetic-induced phase transition: Increased correlation length and emergence of spatial self-organization. AB - In a recent series of papers, the authors have developed a stochastic theory to describe the electrical response of a spatially homogeneous cerebral cortex to infusion of a general anesthetic agent. We showed that by modeling the GABAergic (propofol-like) drug effect as a prolongation of the inhibitory postsynaptic impulse response, we obtain a prediction that there will be a hysteretically separated pair of first-order phase transitions in the population-average excitatory soma voltage, the first occurring at the point of induction of unconsciousness, and the second at the point of emergence from unconsciousness. In the present paper we generalize our earlier "zero-dimensional" homogeneous cortex to a one-dimensional (1D) line of cortical "mass," thus allowing for the possibility of spatial inhomogeneities in neural activity. Following the spirit of our earlier adiabatic ("slow membrane") philosophy, we impose a spatioadiabatic approximation that permits us to compute analytic expressions for changes in EEG (electroencephalographic) correlation length and EEG spatial covariance as a function of anesthetic effect. We establish that the correlation length of the EEG fluctuations is expected to increase at the approach to the transition points, and this finding is consistent with both the homogeneous cortex prediction of increased correlation time ("critical slowing down") near transition, and the recent, comprehensive anesthetic study by John et al. [Conscious. Cogn. 10, 165 (2001)] reporting an increase in EEG coherence near the points of loss and recovery of consciousness. In addition, we find that if the long-range (corticocortical) excitatory-to-inhibitory connectivity in the 1D cortex is stronger than the long-range excitatory-to-excitatory connectivity, then the spatioadiabatic system can organize itself into large-amplitude spatial patterns ("dissipative structures") consisting of giant stationary quasiperiodic voltage fluctuations distributed along the cortical rod. PMID- 14525002 TI - Models of spatial and orientational self-organization of microtubules under the influence of gravitational fields. AB - Tabony and co-workers [C. Papaseit, N. Pochon, and J. Tabony, Proc. Natl. Acad. Sci. U.S.A. 97, 8364 (2000)] showed that the self-organization of microtubules from purified tubulin solutions is sensitive to gravitational conditions. In this paper, we propose two models of spatial and orientational self-organization of microtubules in a gravitational field. First, the spatial model is based on the dominant chemical kinetics. The pattern formation of microtubule concentration is obtained (1) in terms of a moving kink in the limit when the disassembly rate is negligible, and (2) for the case of no free tubulin and only assembled microtubules present. Second, the orientational pattern of striped microtubule domains is consistent with predictions from a phenomenological Landau-Ginzburg free energy expansion in terms of an orientational order parameter. PMID- 14525003 TI - Statistical mechanical approaches to models with many poorly known parameters. AB - Models of biochemical regulation in prokaryotes and eukaryotes, typically consisting of a set of first-order nonlinear ordinary differential equations, have become increasingly popular of late. These systems have large numbers of poorly known parameters, simplified dynamics, and uncertain connectivity: three key features of a class of problems we call sloppy models, which are shared by many other high-dimensional multiparameter nonlinear models. We use a statistical ensemble method to study the behavior of these models, in order to extract as much useful predictive information as possible from a sloppy model, given the available data used to constrain it. We discuss numerical challenges that emerge in using the ensemble method for a large system. We characterize features of sloppy model parameter fluctuations by various spectral decompositions and find indeed that five parameters can be used to fit an elephant. We also find that model entropy is as important to the problem of model choice as model energy is to parameter choice. PMID- 14525004 TI - Dielectric boundary force and its crucial role in gramicidin. AB - In an electrostatic problem with nonuniform geometry, a charge Q in one region induces surface charges [called dielectric boundary charges (DBC)] at boundaries between different dielectrics. These induced surface charges, in return, exert a force [called dielectric boundary force (DBF)] on the charge Q that induced them. The DBF is often overlooked. It is not present in standard continuum theories of (point) ions in or near membranes and proteins, such as Gouy-Chapman, Debye Huckel, Poisson-Boltzmann or Poisson-Nernst- Planck. The DBF is important when a charge Q is near dielectric interfaces, for example, when ions permeate through protein channels embedded in biological membranes. In this paper, we define the DBF and calculate it explicitly for a planar dielectric wall and for a tunnel geometry resembling the ionic channel gramicidin. In general, we formulate the DBF in a form useful for continuum theories, namely, as a solution of a partial differential equation with boundary conditions. The DBF plays a crucial role in the permeation of ions through the gramicidin channel. A positive ion in the channel produces a DBF of opposite sign to that of the fixed charge force (FCF) produced by the permanent charge of the gramicidin polypeptide, and so the net force on the positive ion is reduced. A negative ion creates a DBF of the same sign as the FCF and so the net (repulsive) force on the negative ion is increased. Thus, a positive ion can permeate the channel, while a negative ion is excluded from it. In gramicidin, it is this balance between the FCF and DBF that allows only singly charged positive ions to move into and through the channel. The DBF is not directly responsible, however, for selectivity between the alkali metal ions (e.g., Li+, Na+, K+): we prove that the DBF on a mobile spherical ion is independent of the ion's radius. PMID- 14525005 TI - Rectification efficiency of a Brownian motor. AB - The energy balance of a Brownian motor is discussed based on a Langevin equation without the overdamped approximation. Energetics of the system suggests that the frictional dissipation energy associated with the unidirectional movement should be counted as a part of the useful energy for the rectification process of a Brownian motor. This leads to a new definition of the efficiency, which is applicable, contrary to the conventional one, even if the external load is absent. For the so-called flashing ratchet model, we numerically solve the Langevin equation for various situations and discuss both the temperature and the friction strength dependence of the rectification efficiency and the role of the duty ratio. PMID- 14525006 TI - Kinesin motion in the absence of external forces characterized by interference total internal reflection microscopy. AB - We study the motion of the kinesin molecular motor along microtubules using interference total internal reflection microscopy. This technique achieves nanometer scale resolution together with a fast time response. We describe the first in vitro observation of kinesin stepping at high ATP concentration in the absence of an external load, where the 8-nm step can be clearly distinguished. The short-time resolution allows us to measure the time constant related to the relative motion of the bead-motor connection; we deduce the associated bead-motor elastic modulus. PMID- 14525007 TI - Metachronal waves for deterministic switching two-state oscillators with hydrodynamic interaction. AB - We employ a model system, called rowers, as a generic physical framework to define the problem of the coordinated motion of cilia (the metachronal wave) as a far from equilibrium process. Rowers are active (two-state) oscillators in a low Reynolds number fluid, and interact solely through the forces of hydrodynamic origin. In this work, we consider the case of fully deterministic dynamics, find analytical solutions of the equation of motion in the long-wavelength (continuum) limit, and investigate numerically the short-wavelength limit. We prove the existence of metachronal waves below a characteristic wavelength. Such waves are unstable and become stable only if the sign of the coupling is reversed. We also find that with normal hydrodynamic interaction, the metachronal pattern has the form of stable trains of traveling wave packets sustained by the onset of anti coordinated beating of consecutive rowers. PMID- 14525008 TI - Two-vibron bound states in alpha-helix proteins: the interplay between the intramolecular anharmonicity and the strong vibron-phonon coupling. AB - The influence of the intramolecular anharmonicity and the strong vibron-phonon coupling on the two-vibron dynamics in an alpha-helix protein is studied within a modified Davydov model. The intramolecular anharmonicity of each amide-I vibration is considered, and the vibron dynamics is described according to the small polaron approach. A unitary transformation is performed to remove the intramolecular anharmonicity, and a modified Lang-Firsov transformation is applied to renormalize the vibron-phonon interaction. Then a mean field procedure is realized to obtain the dressed anharmonic vibron Hamiltonian. It is shown that the anharmonicity modifies the vibron-phonon interaction, which results in an enhancement of the dressing effect. In addition, both the anharmonicity and the dressing favor the occurrence of two different bound states whose properties strongly depend on the interplay between the anharmonicity and the dressing. This dependence was summarized in a phase diagram which characterizes the number and the nature of the bound states as a function of the relevant parameters of the problem. For a significant anharmonicity, the low-frequency bound states describe two vibrons trapped onto the same amide-I vibration, whereas the high-frequency bound states refer to the trapping of the two vibrons onto nearest neighbor amide I vibrations. PMID- 14525009 TI - Totally asymmetric exclusion process with extended objects: a model for protein synthesis. AB - The process of protein synthesis in biological systems resembles a one dimensional driven lattice gas in which the particles have spatial extent, covering more than one lattice site. We expand the well studied totally asymmetric exclusion process, in which particles typically cover a single lattice site, to include cases with extended objects. Exact solutions can be determined for a uniform closed system. We analyze the uniform open system through two approaches. First, a continuum limit produces a modified diffusion equation for particle density profiles. Second, an extremal principle based on domain wall theory accurately predicts the phase diagram and currents in each phase. Finally, we briefly consider approximate approaches to a nonuniform open system with quenched disorder in the particle hopping rates and compare these approaches with Monte Carlo simulations. PMID- 14525010 TI - Modeling DNA structure, elasticity, and deformations at the base-pair level. AB - We present a generic model for DNA at the base-pair level. We use a variant of the Gay-Berne potential to represent the stacking energy between the neighboring base pairs. The sugar-phosphate backbones are taken into account by semirigid harmonic springs with a nonzero spring length. The competition between these two interactions and the introduction of a simple geometrical constraint lead to a stacked right-handed B-DNA-like conformation. The mapping of the presented model to the Marko-Siggia and the stack-of-plates model enables us to optimize the free model parameters so as to reproduce the experimentally known observables such as persistence lengths, mean and mean-squared base-pair step parameters. For the optimized model parameters, we measured the critical force where the transition from B- to S-DNA occurs to be approximately 140 pN. We observe an overstretched S DNA conformation with highly inclined bases which partially preserves the stacking of successive base pairs. PMID- 14525011 TI - Fokker-Planck perspective on stochastic delay systems: exact solutions and data analysis of biological systems. AB - Stochastic delay systems with additive noise are examined from the perspective of Fokker-Planck equations. For a linear system, the exact stationary probability density is derived by means of a delay Fokker-Planck equation. We show how to determine the delay equation of the linear system from experimental data, and corroborate a fundamental result previously obtained by Kuchler and Mensch. We also propose a method to derive delay equations of nonlinear systems from experimental data. To this end, the theory of multivariate Fokker-Planck equations is used. The applicability of this method is demonstrated for stochastic models describing tracking and pointing movements of humans. PMID- 14525012 TI - Multifractal and correlation analyses of protein sequences from complete genomes. AB - A measure representation of protein sequences similar to the measure representation of DNA sequences proposed in our previous paper [Yu et al., Phys. Rev. E 64, 031903 (2001)] and another induced measure are introduced. Multifractal analysis is then performed on these two kinds of measures of a large number of protein sequences derived from corresponding complete genomes. From the values of the D(q) (generalized dimensions) spectra and related C(q) (analogous specific heat) curves, it is concluded that these protein sequences are not completely random sequences. For substrings with length K=5, the D(q) spectra of all organisms studied are multifractal-like and sufficiently smooth for the C(q) curves to be meaningful. The C(q) curves of all bacteria resemble a classical phase transition at a critical point. But the "analogous" phase transitions of higher organisms studied exhibit the shape of double-peaked specific heat function. But for the classification problem, the multifractal property is not sufficient. When the measure representations of protein sequences from complete genomes are considered as time series, a method based on correlation analysis after removing some memory from the time series is proposed to construct a phylogenetic tree. This construction is shown to be reasonably satisfactory. PMID- 14525013 TI - Spatiotemporal dynamics of optical molecular motors. AB - The spatiotemporal dynamics of optical molecular motors is simulated on the basis of a spatially resolved model. A spatially dependent Fokker-Planck model for the molecular motors is linked with Maxwell's wave equation describing the external excitation via a spatially inhomogeneous light field. Simulations show that strong diffusion of the embedding fluent leads to increased motor dynamics while in inhomogeneous ensembles motor clustering may occur. Spatially inhomogeneous optical excitation may provide a means of movement control of the molecular motors. PMID- 14525014 TI - Sparks and waves in a stochastic fire-diffuse-fire model of Ca2+ release. AB - Calcium ions are an important second messenger in living cells. Indeed, calcium signals in the form of waves have been the subject of much recent experimental interest. It is now well established that these waves are composed of elementary stochastic release events (calcium puffs or sparks) from spatially localized calcium stores. Here we develop a computationally inexpensive model of calcium release, based upon a stochastic generalization of the fire-diffuse-fire threshold model. Our model retains the discrete nature of calcium stores, but also incorporates a notion of release probability via the introduction of threshold noise. Numerical simulations of the model illustrate that stochastic calcium release leads to the spontaneous production of calcium sparks that may merge to form saltatory waves. In the parameter regime where deterministic waves exist, it is possible to identify a critical level of noise, defining a nonequilibrium phase transition between propagating and abortive structures. A statistical analysis shows that this transition is the same as for models in the directed percolation universality class. Moreover, in the regime where no initial structure can survive deterministically, threshold noise is shown to generate a form of array enhanced coherence resonance, whereby all calcium stores release periodically and simultaneously. PMID- 14525015 TI - Coherent states of Gompertzian growth. AB - The origin of the Gompertz function G(t)=G(0)e(b/a(1-e(-at))) widely applied to fit the biological and medical data, particularly growth of organisms, organs, and tumors is analyzed. It is shown that this function is a solution of a time dependent counterpart of the Schrodinger equation for the Morse oscillator with anharmonicity constant equal to 1. The coherent states of the Gompertzian systems, which minimize the time-energy uncertainty relation, have been found. These are eigenstates of the annihilation operator identified with the operator of growth, whereas eigenstates of the creation operator represent the Gompertzian states of regression. The coherent formation of the specific growth patterns in the Gompertzian systems appears as a result of the nonlocal long-range cooperation between the microlevel (the individual cell) and the macrolevel (the system as a whole). PMID- 14525016 TI - Spiral wave stability in cardiac tissue with biphasic restitution. AB - Human ventricular tissue as well as several animal ventricular preparations show a biphasic shape of the action potential duration restitution curve, with a local maximum at low diastolic intervals. We study numerically how the location and properties of this nonmonotonicity affect the stability of spiral waves. We find that, depending on the slopes of the ascending and of the descending parts of the restitution curve, we can have either stable rotation of the spiral wave or spiral breakup. We identify two types of spiral breakup: one due to a steep positive slope and another due to a steep negative slope in the restitution curve. We discuss the differences in their manifestation and possible implications. We also find that increasing the slope of the descending part of the restitution curve increases the meandering of the spiral wave, due to the repeated occurrence of conduction blocks near the spiral wave tip. PMID- 14525017 TI - Characterization of flow reduction properties in an aneurysm due to a stent. AB - We consider a lattice Boltzmann simulation of blood flow in a vessel deformed by the presence of an aneurysm. Modern clinical treatments involve introducing a stent (a tubular mesh of wires) into the cerebral artery in order to reduce the flow inside the aneurysm and favor its spontaneous reabsorption. A crucial question is to design the stent with suitable porosity so as to produce the most effective flow reduction. We propose a stent positioning factor as a characterizing tool for stent pore design in order to describe the flow reduction effect and reveal the several flow reduction mechanisms using this effect. PMID- 14525018 TI - Slow switching in a population of delayed pulse-coupled oscillators. AB - We show that peculiar collective dynamics called slow switching arises in a population of leaky integrate-and-fire oscillators with delayed, all-to-all pulse couplings. By considering the stability of cluster states and symmetry possessed by our model, we argue that saddle connections between a pair of the two-cluster states are formed under general conditions. Slow switching appears as a result of the system's approach to the saddle connections. It is also argued that such saddle connections are easy to arise near the bifurcation point where the state of perfect synchrony loses stability. We develop an asymptotic theory to reduce the model into a simpler form, with which an analytical study of the cluster states becomes possible. PMID- 14525019 TI - Firing statistics of a neuron model driven by long-range correlated noise. AB - We study the statistics of the firing patterns of a perfect integrate and fire neuron model driven by additive long-range correlated Ornstein-Uhlenbeck noise. Using a quasistatic weak noise approximation we obtain expressions for the interspike interval (ISI) probability density, the power spectral density, and the spike count Fano factor. We find unimodal, long-tailed ISI densities, Lorenzian power spectra at low frequencies, and a minimum in the Fano factor as a function of counting time. The implications of these results for signal detection are discussed. PMID- 14525020 TI - Structure, dynamics, and energetics of water at the surface of a small globular protein: a molecular dynamics simulation. AB - The dynamics of water around a biomolecular surface has attracted a lot of attention recently. We report here protein-solvent simulation studies of the small globular protein ubiquitin (human). The simulations are run unconstrained, without freezing the bonds. The mean square displacements of the water oxygen atoms show a sublinear trend with time. The diffusion coefficient data indicate that the water in the first hydration layer behaves like water at a temperature that is roughly 12 degrees C lower than the average temperature of the system (27 degrees C). Both the dipolar second-rank relaxation and the survival time correlation function of the water layers show two decay constants, indicating contributions from fast and slow dynamics. A calculation of the interaction energy between the water layers and protein indicates that the interaction energy sharply decreases beyond 4 A from the protein surface. PMID- 14525021 TI - Nonuniform corticothalamic continuum model of electroencephalographic spectra with application to split-alpha peaks. AB - Recent theoretical work has successfully predicted electroencephalographic spectra from physiology using a model corticothalamic system with spatially uniform parameters. The present work incorporates parameter nonuniformities into this model via the coupling they induce between spatial eigenmodes. Splitting of the spectral alpha peak, an effect seen in a small percentage of the normal population, is investigated as an illustrative special case. It is confirmed that weak splitting can arise from mode structure if the peak is sufficiently sharp, even for uniform parameters. However, it is further demonstrated that greater splitting can result from nonuniformities, and it is argued that this mechanism for split alpha is better able to account quantitatively for this effect than previously suggested alternatives of pacemakers or purely cortical resonances. On introducing nonuniformities in corticothalamic loop time delays, we find that the alpha frequency also varies as one moves from the front to the back of the head, in accord with observations, and that analogous (but less distinct) variations are seen in the beta peak. Analysis shows realistic variations of around +/-10 ms relative to the mean loop delay of approximately 80 ms can account for observed splittings of about 1 Hz. It is also suggested that subjects who display clear alpha splitting form the tail of a distribution of magnitude of cortical inhomogeneity, rather than a separate population. PMID- 14525022 TI - Stability of negative-image equilibria in spike-timing-dependent plasticity. AB - We investigate the stability of negative image equilibria in mean synaptic weight dynamics governed by spike-timing-dependent plasticity (STDP). The model architecture closely follows the anatomy and physiology of the electrosensory lateral line lobe (ELL) of mormyrid electric fish. The ELL uses a spike-timing dependent learning rule to form a negative image of the reafferent signal from the fish's own electric discharge, thus improving detectability of external electric fields. We derive sufficient conditions for existence of the negative image and necessary and sufficient conditions for stability, for arbitrary postsynaptic potential functions and arbitrary learning rules. This significantly generalizes earlier investigations. We then apply the general result to several examples of biological interest, including a class of learning rules consistent with the rule observed experimentally in the mormyrid ELL. PMID- 14525023 TI - Compressing inverse lyotropic systems: Structural behavior and energetics of dioleoyl phosphatidyl ethanolamine. AB - The pressure effects on the stability and energetics of lipid phases in the L alpha-dioleoyl phosphatidyl ethanolamine (DOPE)-water system are presented. Using synchrotron diffraction experiments, performed at a wide range of concentrations, pressure-induced transitions from the inverse hexagonal (H(II)) to the lamellar L(alpha) phase and from the L(alpha) to the lamellar L(beta) phase are demonstrated. Moreover, in the most dehydrated samples an intermediate phase is found between the H(II) and the L(alpha) phases, confirming that the lamellar-to nonlamellar phase transition occurs through key intermediate structures. Simple molecular packing arguments lead to an interpretation of the phase behavior: in fact, pressure induces a progressive stiffening of the DOPE hydrocarbon chains and a reduction of the cross-sectional area. Because pressure is more effective in reducing the cross-sectional area near the terminal methyl groups than at the water-lipid interface, the curvature of that interface in the H(II) phase is reduced during compression. The work of isothermal compression was then obtained and analyzed in terms of the elastic energetic contributions that should stabilize the DOPE phases during compression. As a result, we observe that the isothermal lateral compression modulus is almost independent of concentration, but it increases as a function of pressure, suggesting that the DOPE repulsion becomes very strong while the whole lipid shape becomes more cylindrical. On the other hand, the bending rigidity is observed to decrease with increasing pressure, while the spontaneous curvature becomes less negative. This suggests that the chain repulsion becomes relatively weaker, and thus less efficient in balancing the torque of head-group repulsion, as the order parameter increases. PMID- 14525024 TI - Interplay of chemotaxis and chemokinesis mechanisms in bacterial dynamics. AB - Motivated by observations of the dynamics of Myxococcus xanthus, we present a self-interacting random walk model that describes the competition between chemokinesis and chemotaxis. Cells are constrained to move in one dimension, but release a chemical chemoattractant at a steady state. The bacteria sense the chemical that they produce. The probability of direction reversals is modeled as a function of both the absolute level of chemoattractant sensed directly under each cell as well as the gradient sensed across the length of the cell. If the chemical does not degrade or diffuse rapidly, the one-dimensional trajectory depends on the entire past history of the trajectory. We derive the corresponding Fokker-Planck equations, use an iterative mean-field approach that we solve numerically for short times, and perform extensive Monte Carlo simulations of the model. Cell positional distributions and the associated moments are computed in this feedback system. Average drift and mean squared displacements are found. Crossover behaviors among different diffusion regimes are found. PMID- 14525025 TI - Competing reactions with initially separated components in the asymptotic time region. AB - Two competing irreversible reactions with initially separated components and with essentially different reaction constants are theoretically studied in the asymptotic time region. The description of the two simultaneous reactions is reduced to the consideration of two reactions separated in space. It is shown that the reaction rate profile can have two maxima and their ratio is independent of time. The location and relative value of the maxima are functions of the reaction constants and initial concentrations. PMID- 14525026 TI - Transition from stable to unstable growth by an inertial force. AB - We introduce a simple growth model where the growth of the interface is affected by an inertial force and a white noise. The magnitude of the inertial force is controlled by a constant p between 0 and 1. An inertial force increases continuously from 0, as p does from 0 to 1. In our model, the interface starts growing from a flat state. When pp(c), however, the interface width increases continuously without saturation as time elapses. We explain via simple calculation how this interesting phenomenon occurs in our model. We find p(c)=0.5 from the calculation. This critical value is in excellent agreement with the critical value p(c)=0.50(1) found from the simulations of our model. PMID- 14525027 TI - Scaling function for surface width for free boundary conditions. AB - We study the restricted curvature model with both periodic and free boundary conditions and show that the scaling function of the surface width depends on the type of boundary conditions. When the free boundary condition is applied, the surface width shows a new dynamic scaling whose asymptotic behavior is different from the usual scaling behavior of the self-affine surfaces. We propose a generalized scaling function for the surface width for free boundary conditions and introduce a normalized surface width to clarify the origin of the superrough phenomena of the model. PMID- 14525028 TI - Apparent low-field spin-lattice dispersion in the smectic-A mesophase of thermotropic cyanobiphenyls. AB - Proton field-cycling spin-lattice relaxometry T1 of the smectic-A mesophase in cyanobiphenyls revealed the presence of steep dispersions in the low-frequency regime. We clearly show that the strong dispersion characteristic of smectic organizations cannot be attributed to the collective molecular dynamics (order director fluctuations), as it is usually interpreted. We present two independent experimental evidences: the dependence of the dispersion with the slew rate of the magnetic field cycle and the dependence of the dispersion with the presence and power of an ultrasonic field. PMID- 14525029 TI - Comment on "Optical determination of flexoelectric coefficients and surface polarization in a hybrid aligned nematic cell". AB - We argue that the experimental determination of the flexoelectric coefficients and the surface polarization of the p-pentyl-cyanobiphenil liquid crystal, recently reported by Mazzulla et al. [Phys. Rev. E 64, 021708 (2001)] is questionable. This conclusion follows from the hypothesis performed by the authors to extract the flexoelectric coefficients and the surface polarization from the reflectivity curves taken under an applied voltage. In fact, they assume that the liquid crystal can be considered as a perfect insulating material and that the ionic charges do not contribute to the electric field profile inside the sample. Using our recent results concerning nematic liquid crystals submitted to an external electric field we show that this hypothesis is far from being correct. In the experimental conditions of Mazzulla et al., due to the low frequency of the applied alternative square pulsed signal, the ions play an important role in the molecular orientation of the liquid crystal induced by the external field. PMID- 14525031 TI - Comment on "Molecular gyroscopes and biological effects of weak extremely low frequency magnetic fields". AB - A mechanism whereby reaction rates may be influenced by weak alternating magnetic fields has been suggested by Binhi and Savin [Phys. Rev. E 65, 051912 (2002)] to account for certain magnetobiological effects. It is proposed that the fields influence the probability of reaction of molecular rotators (gyroscopes) by inducing interference between eigenstates of angular momentum superposed in their wave functions. The predicted variation of reaction rate with the amplitude of the alternating field is found to be qualitatively consistent with observation. It is commented that the required interference occurs only in circumstances which are quite implausible, and that even if it were possible, the interference would not lead to a detectable magnetobiological effect. PMID- 14525033 TI - Nonuniversality of elastic exponents in random bond-bending networks. AB - We numerically investigate the rigidity percolation transition in two-dimensional flexible, random rod networks with freely rotating cross links. Near the transition, networks are dominated by bending modes and the elastic modulii vanish with an exponent f=3.0+/-0.2, in contrast with central force percolation which shares the same geometric exponents. This indicates that universality for geometric quantities does not imply universality for elastic ones. The implications of this result for actin-fiber networks is discussed. PMID- 14525034 TI - Emergence of pulled fronts in fermionic microscopic particle models. AB - We study the emergence and dynamics of pulled fronts described by the Fisher Kolmogorov-Petrovsky-Piscounov (FKPP) equation in the microscopic reaction diffusion process A+A<-->A on the lattice when only a particle is allowed per site. To this end we identify the parameter that controls the strength of internal fluctuations in this model, namely, the number of particles per correlated volume. When internal fluctuations are suppressed, we explictly see the matching between the deterministic FKPP description and the microscopic particle model. PMID- 14525035 TI - Galilean-invariant lattice-Boltzmann models with H theorem. AB - We demonstrate that the requirement of Galilean invariance determines the choice of H function for a wide class of entropic lattice-Boltzmann models for the incompressible Navier-Stokes equations. The required H function has the form of the Burg entropy for D=2, and of a Tsallis entropy with q=1-(2/D) for D>2, where D is the number of spatial dimensions. We use this observation to construct a fully explicit, unconditionally stable, Galilean-invariant, lattice-Boltzmann model for the incompressible Navier-Stokes equations, for which attainable Reynolds number is limited only by grid resolution. PMID- 14525036 TI - Periodic phase synchronization in coupled chaotic oscillators. AB - We investigate the characteristics of temporal phase locking states observed in the route to phase synchronization. It is found that before phase synchronization there is a periodic phase synchronization state characterized by periodic appearance of temporal phase-locking state and that the state leads to local negativeness in one of the vanishing Lyapunov exponents. By taking a statistical measure, we present the evidences of the phenomenon in unidirectionally and mutually coupled chaotic oscillators, respectively. And it is qualitatively discussed that the phenomenon is described by a nonuniform oscillator model in the presence of noise. PMID- 14525037 TI - Experimental Chua-plasma phase synchronization of chaos. AB - Experimental phase synchronization of chaos is demonstrated for two different chaotic oscillators: a plasma discharge and the Chua circuit. Our technique includes real-time capability for observing synchronization-desynchronization transitions. This capability results from a strong combination of synchronization and control, and allows tuning adjustments for search and stabilization of synchronous states. A power law is observed for the mean time between 2pi phase slips for different coupling strenghts. The experimental results are consistent with the numerical simulations. PMID- 14525038 TI - Large particle number limit in rain. AB - The way we conceptualize rain is fundamental in many branches of science since it provides the basis not only for rain modeling notably in meteorology and hydrology, but also for interpreting rain data (from gauges and radars). In order to empirically address this question, we use stereophotographic data to measure the positions and volumes of raindrops from approximately 10 m(3) regions containing 5000-15,000 of these drops. By determining the drop statistics in spheres of increasing size, we conduct a basic continuum mechanics thought experiment. We show that-presumably due to turbulence-there is no microscale macroscale separation. We find that the large particle number (N) limit in rain is not a homogeneous continuum, but rather it is nonclassical, strongly inhomogeneous, and approaching a multifractal discontinuum. PMID- 14525039 TI - Unstable periodic solutions embedded in a shell model turbulence. AB - An approach to intermittency of a shell model turbulence is proposed from the viewpoint of dynamical systems. We detected unstable solutions of the Gledzer Ohkitani-Yamada shell model and studied their relation to turbulence statistics. One of the solutions has an unstable periodic orbit (UPO), which shows an intermittency where the scaling exponents of the structure function have a nonlinear dependence on its order, quite similar to that of turbulence solution at the same parameter values. The attractor in the phase space is found to be well approximated by a continuous set of solutions generated from the UPO through a one-parameter phase transformation, which implies that the intermittency of the shell model turbulence is described by this UPO. PMID- 14525040 TI - Simple model for drag reduction. AB - Direct numerical simulations established that the finite-extension nonlinear elasticity-Peterlin (FENE-P) model of viscoelastic flows exhibits the phenomenon of turbulent drag reduction which is caused in experiments by dilute polymeric additives. To gain analytic understanding of the phenomenon, we introduce in this paper a simple one-dimensional model of the FENE-P equations. We demonstrate drag reduction in the simple model, and explain analytically the main observations which include (i) reduction of velocity gradients for fixed throughput and (ii) increase of throughput for fixed dissipation. PMID- 14525041 TI - Self-trapped bidirectional waveguides in a saturable photorefractive medium. AB - A time-dependent model for the generation of joint waveguides by counterpropagating light beams in photorefractive crystals is introduced. Depending on initial conditions and parameter values, the beams form stable structures or display periodic and irregular dynamics. Steady-state solutions nonuniform in the direction of propagation are found, representing a general class of self-trapped waveguides that include counterpropagating spatial vector solitons as a particular case. PMID- 14525042 TI - Optical modulational instability in a nonlocal medium. AB - We report the first observation of modulational instability in nematic liquid crystals encompassing a nonlocal behavior. The experimental results are quantitatively compared with the theory revealing the fundamental features associated to a nonlocal nonlinearity. PMID- 14525043 TI - Peculiarities of first-order phase transitions in the presence of an electric field. AB - In this study, we developed a variational approach for thermodynamic systems with nondistributed parameters in the presence of the external electrostatic field. Using the obtained general relations, we analyzed some characteristic features of the first-order phase transitions in the presence of electric field. We determined the range of the thermodynamic parameters where both phases are stable (hysteresis), and the range of the parameters whereby both phases are metastable. In the range where both phases are metastable, we considered kinetics of formation of a new phase and determined the dependencies of the concentrations of phases in the region of their metastability on the amplitude of the external electric field. The obtained results imply the feasibility to control phase composition in the system by varying the amplitude of the external electrostatic field. PMID- 14525044 TI - Dynamical model of a cooperative driving system for freeway traffic. AB - We propose an extended optimal velocity model applicable to cooperative driving control system, which will be realized in the near future. In the model, a vehicle is controlled by the system using the information of arbitrary number of vehicles that precede or follow. We investigate the stability of uniform flow and the response to a disturbance in the linear approximation. PMID- 14525045 TI - Traveling front solutions to directed diffusion-limited aggregation, digital search trees, and the Lempel-Ziv data compression algorithm. AB - We use the traveling front approach to derive exact asymptotic results for the statistics of the number of particles in a class of directed diffusion-limited aggregation models on a Cayley tree. We point out that some aspects of these models are closely connected to two different problems in computer science, namely, the digital search tree problem in data structures and the Lempel-Ziv algorithm for data compression. The statistics of the number of particles studied here is related to the statistics of height in digital search trees which, in turn, is related to the statistics of the length of the longest word formed by the Lempel-Ziv algorithm. Implications of our results to these computer science problems are pointed out. PMID- 14525046 TI - Clustering properties of a generalized critical Euclidean network. AB - Many real-world networks exhibit a scale-free feature, have a small diameter, and a high clustering tendency. We study the properties of a growing network, which has all these features, in which an incoming node is connected to its ith predecessor of degree k(i) with a link of length l using a probability proportional to k(beta)(i)l(alpha). For alpha>-0.5, the network is scale-free at beta=1 with the degree distribution P(k) proportional to k(-gamma) and gamma=3.0 as in the Barabasi-Albert model (alpha=0,beta=1). We find a phase boundary in the alpha-beta plane along which the network is scale-free. Interestingly, we find a scale-free behavior even for beta>1 for alpha<-0.5, where the existence of a different universality class is indicated from the behavior of the degree distribution and the clustering coefficients. The network has a small diameter in the entire scale-free region. The clustering coefficients emulate the behavior of most real networks for increasing negative values of alpha on the phase boundary. PMID- 14525047 TI - Number fluctuation and the fundamental theorem of arithmetic. AB - We consider N bosons occupying a discrete set of single-particle quantum states in an isolated trap. Usually, for a given excitation energy, there are many combinations of exciting different number of particles from the ground state, resulting in a fluctuation of the ground state population. As a counterexample, we take the quantum spectrum to be logarithms of the prime number sequence, and using the fundamental theorem of arithmetic, find that the ground state fluctuation vanishes exactly for all excitations. The use of the canonical or grand canonical ensembles, on the other hand, gives a substantial number fluctuation for the ground state. This is an example of a system where canonical and grand canonical ensemble averagings are not valid because of the peculiar nature of the quantum spectrum. PMID- 14525048 TI - Self-avoiding walks and connective constants in small-world networks. AB - Long-distance characteristics of small-world networks have been studied by means of self-avoiding walks (SAW's). We consider networks generated by rewiring links in one- and two-dimensional regular lattices. The number of SAW's u(n) was obtained from numerical simulations as a function of the number of steps n on the considered networks. The so-called connective constant, mu=lim(n- >infinity)u(n)/u(n-1), which characterizes the long-distance behavior of the walks, increases continuously with disorder strength (or rewiring probability p). For small p, one has a linear relation mu=mu(0)+ap, mu(0) and a being constants dependent on the underlying lattice. Close to p=1 one finds the behavior expected for random graphs. An analytical approach is given to account for the results derived from numerical simulations. Both methods yield results agreeing with each other for small p, and differ for p close to 1, because of the different connectivity distributions resulting in both cases. PMID- 14525050 TI - Partition functions and metropolis-type evolution rules for surface growth models with constraints. AB - We study dynamical scaling properties of the surface growth model with the Metropolis-type evolution rule from a partition function Z= sum ([h(r)])II (h(max))(h=h(min))1/2(1+z(n(h))), where z is a fugacity-like quantity and n(h) is the number of sites with height h in a surface configuration [h(r)]. The partition function describes a 2-particle correlated growth model when z=-1 and a self-flattening growth model when z=0. For one-dimensional equilibrium surfaces, the scaling properties for z>or=-1 except z=1 are all one phase with roughness exponent alpha=1/3 and growth exponent beta approximately equal 0.22. For the growing (eroding) surfaces, there exists a phase transition at z=0 from the grooved phase (alpha=1) for -10. PMID- 14525049 TI - Properties of random graphs with hidden color. AB - We investigate in some detail a recently suggested general class of ensembles of sparse undirected random graphs based on a hidden stub coloring, with or without the restriction to nondegenerate graphs. The calculability of local and global structural properties of graphs from the resulting ensembles is demonstrated. Cluster size statistics are derived with generating function techniques, yielding a well-defined percolation threshold. Explicit rules are derived for the enumeration of small subgraphs. Duality and redundancy is discussed, and subclasses corresponding to commonly studied models are identified. PMID- 14525051 TI - Thermodynamic instabilities in one-dimensional particle lattices: a finite-size scaling approach. AB - One-dimensional thermodynamic instabilities are phase transitions, not prohibited by Landau's argument because the energy of the domain wall which separates the two phases is infinite. Whether they actually occur in a given system of particles must be demonstrated on a case-by-case basis by examining the properties of the corresponding singular transfer integral (TI) equation. The present work deals with the generic Peyrard-Bishop model of DNA denaturation. In the absence of exact statements about the spectrum of the singular TI equation, I use Gauss-Hermite quadratures to achieve a single-parameter-controlled approach to rounding effects; this allows me to employ finite-size scaling concepts in order to demonstrate that a phase transition occurs and to derive the critical exponents. PMID- 14525052 TI - Effects of random noise on a simple class of growing network models. AB - We investigate the effects of random noise on network systems. In particular, we consider a simple class of growing network models whose topological structure is determined by the preferred attachment A(k). We introduce a noise-induced attachment A(k) which includes fluctuations in the number of links of individual nodes due to a random noise. We carry out the numerical simulations to show that the topological structure of the networks is determined not only by A(k) but also by the strength of the noise. Analytic and numerical solutions are also presented to support this observation. In addition, we study the stability of networks against attacks under the noisy condition. Similarly, we introduce a noise induced preferred deletion B(k), and show that noise is an essential feature to determine the stability of networks. PMID- 14525053 TI - Damped harmonic oscillator: pure states of the bath and exact master equations. AB - Time evolution of a harmonic oscillator linearly coupled to a heat bath is compared for three classes of initial states for the bath modes-grand canonical ensemble, number states, and coherent states. It is shown that for a wide class of number states the behavior of the oscillator is similar to the case of the equilibrium bath. If the bath modes are initially in coherent states, then the variances of the oscillator coordinate and momentum, as well as its entanglement to the bath, asymptotically approach the same values as for the oscillator at zero temperature and the average coordinate and momentum show a Brownian-like behavior. We derive an exact master equation for the characteristic function of the oscillator valid for arbitrary factorized initial conditions. In the case of the equilibrium bath this equation reduces to an equation of the Hu-Paz-Zhang type, while for the coherent states bath it leads to an exact stochastic master equation with a multiplicative noise. PMID- 14525054 TI - Origin of degree correlations in the Internet and other networks. AB - It has been argued that the observed anticorrelation between the degrees of adjacent vertices in the network representation of the Internet has its origin in the restriction that no two vertices have more than one edge connecting them. Here, we propose a formalism for modeling ensembles of graphs with single edges only and derive values for the exponents and correlation coefficients characterizing them. Our results confirm that the conjectured mechanism does indeed give rise to correlations of the kind seen in the Internet, although only a part of the measured correlation can be accounted for in this way. PMID- 14525056 TI - Full reduction of large finite random Ising systems by real space renormalization group. AB - We describe how to evaluate approximately various physical interesting quantities in random Ising systems by direct renormalization of a finite system. The renormalization procedure is used to reduce the number of degrees of freedom to a number that is small enough, enabling direct summing over the surviving spins. This procedure can be used to obtain averages of functions of the surviving spins. We show how to evaluate averages that involve spins that do not survive the renormalization procedure. We show, for the random field Ising model, how to obtain Gamma(r)=-, the "connected" correlation function, and S(r)=, the "disconnected" correlation function. Consequently, we show how to obtain the average susceptibility and the average energy. For an Ising system with random bonds and random fields, we show how to obtain the average specific heat. We conclude by presenting our numerical results for the average susceptibility and the function Gamma(r) along one of the principal axes. (In this work, the full three-dimensional (3D) correlation is calculated and not just parameters such nu or eta). The results for the average susceptibility are used to extract the critical temperature and critical exponents of the 3D random field Ising system. PMID- 14525055 TI - Citation networks in high energy physics. AB - The citation network constituted by the SPIRES database is investigated empirically. The probability that a given paper in the SPIRES database has k citations is well described by simple power laws, P(k) proportional to k(-alpha), with alpha approximately 1.2 for k less than 50 citations and alpha approximately 2.3 for 50 or more citations. A consideration of citation distribution by subfield shows that the citation patterns of high energy physics form a remarkably homogeneous network. Further, we utilize the knowledge of the citation distributions to demonstrate the extreme improbability that the citation records of selected individuals and institutions have been obtained by a random draw on the resulting distribution. PMID- 14525057 TI - Strategy updating rules and strategy distributions in dynamical multiagent systems. AB - In the evolutionary version of the minority game, agents update their strategies (gene value p) in order to improve their performance. Motivated by the recent intriguing results obtained for prize-to-fine ratios, which are smaller than unity, we explore the system's dynamics with a strategy updating rule of the form p-->p+/-delta(p) (0 approximately N(n-g). However, many natural networks have a non-Poissonian degree distribution. Here we present approximate equations for the average number of subgraphs in an ensemble of random sparse directed networks, characterized by an arbitrary degree sequence. We find scaling rules for the commonly occurring case of directed scale-free networks, in which the outgoing degree distribution scales as P(k) approximately k(-gamma). Considering the power exponent of the degree distribution, gamma, as a control parameter, we show that random networks exhibit transitions between three regimes. In each regime, the subgraph number of appearances follows a different scaling law, approximately Nalpha, where alpha=n-g+s-1 for gamma<2, alpha=n g+s+1-gamma for 2gamma(c), where s is the maximal outdegree in the subgraph, and gamma(c)=s+1. We find that certain subgraphs appear much more frequently than in Erdos networks. These results are in very good agreement with numerical simulations. This has implications for detecting network motifs, subgraphs that occur in natural networks significantly more than in their randomized counterparts. PMID- 14525070 TI - Connectivity distribution of spatial networks. AB - We study spatial networks constructed by randomly placing nodes on a manifold and joining two nodes with an edge whenever their distance is less than a certain cutoff. We derive the general expression for the connectivity distribution of such networks as a functional of the distribution of the nodes. We show that for regular spatial densities, the corresponding spatial network has a connectivity distribution decreasing faster than an exponential. In contrast, we also show that scale-free networks with a power law decreasing connectivity distribution are obtained when a certain information measure of the node distribution (integral of higher powers of the distribution) diverges. We illustrate our results on a simple example for which we present simulation results. Finally, we speculate on the role played by the limiting case P(k) proportional, variant k( 1) which appears empirically to be relevant to spatial networks of biological origin such as the ones constructed from gene expression data. PMID- 14525071 TI - Anisotropy effect on two-dimensional cellular-automaton traffic flow with periodic and open boundaries. AB - Using computer simulations we investigate, in a version of the Biham-Middleton Levine model with random sequential update on a square lattice, the anisotropy effect of the probabilities of the change of the motion directions of cars, from up to right (p(ur)) and from right to up (p(ru)), on the dynamical jamming transition and velocities under periodic boundaries on one hand and the phase diagram under open boundaries on the other hand. However, in the former case, the sharp jamming transition appears only for p(ur)=0=p(ru)=0 (i.e., when the cars alter their motion directions). In the open boundary conditions, it is found that the first-order line transition between jamming and moving phases is curved. Hence, by increasing the anisotropy, the moving phase region expands as well as the contraction of the jamming and maximal current phases takes place. Moreover, in the anisotropic case, the transition between the jamming phase (or moving phase) and the maximal current phase is of second order while in the isotropic case, and when each car changes its direction of motion at every time step (p(ru)=p(ur)=1), the transition is of first order. Furthermore, in the maximal current phase, the density profile decays with an exponent gamma approximately 1/4. PMID- 14525072 TI - Counting multiple solutions in glassy random matrix models. AB - This is a first step in counting the number of multiple solutions in certain glassy random matrix models introduced by N. Deo [Phys. Rev. E 65, 056115 (2002)]. We are able to do this by reducing the problem of counting the multiple solutions to that of a moment problem. More precisely, we count the number of different moments when we introduce an asymmetry (tapping) in the random matrix model and then take it to vanish. It is shown here that the number of moments grows exponentially with respect to N, the size of the matrix. As these models map onto models of structural glasses in the high temperature phase (liquid), this may have interesting implications for the supercooled liquid phase in these spin glass models. Further, it is shown that the nature of the asymmetry (tapping) is crucial in finding the multiple solutions. This also clarifies some of the puzzles raised by E. Brezin and N. Deo [Phys. Rev. E 59, 3901 (1999)]. PMID- 14525073 TI - Numerical study of the Sherrington-Kirkpatrick model in a magnetic field. AB - We study numerically the Sherrington-Kirkpatrick model as a function of the magnetic field h, with fixed temperature T=0.6T(c). We investigate the finite size scaling behavior of several quantities, such as the spin-glass susceptibility, searching for numerical evidences of the transition on the de Almeida-Thouless line. We find strong corrections to scaling which make difficult to locate the transition point. This shows, in a simple case, the extreme difficulties of spin-glass simulations in a nonzero magnetic field. Next, we study various sum rules (consequences of stochastic stability) involving overlaps between three and four replicas, which appear to be numerically well satisfied, and in a nontrivial way. Finally, we present data on P(q) for a large lattice size (N=3200) at low temperature T=0.4T(c), where the shape predicted by the replica symmetry breaking solution of the model for a nonzero magnetic field is visible. PMID- 14525074 TI - Correlations, soliton modes, and non-Hermitian linear mode transmutation in the one-dimensional noisy Burgers equation. AB - Using the previously developed canonical phase space approach applied to the noisy Burgers equation in one dimension, we discuss in detail the growth morphology in terms of nonlinear soliton modes and superimposed linear modes. We moreover analyze the non-Hermitian character of the linear mode spectrum and the associated dynamical pinning, and mode transmutation from diffusive to propagating behavior induced by the solitons. We discuss the anomalous diffusion of growth modes, switching and pathways, correlations in the multisoliton sector, and in detail the correlations and scaling properties in the two-soliton sector. PMID- 14525075 TI - Dipolar effective interaction in a fluid of charged spheres near a dielectric plate. AB - Static correlations in a classical fluid of charged spheres at equilibrium are studied in the vicinity of an insulating wall characterized by its dielectric constant. It is well known that the deformations of screening clouds induced by the presence of the wall result into an effective f(alphaalpha('))(x,x('))/y(3) interaction in the pair distribution function between two charges e(alpha) and e(alpha(')) located at distances x and x(') from the wall and separated by a large distance y along the wall. We investigate the structure of f(alphaalpha('))(x,x(')). The method is based on systematic resummations in the Mayer diagrammatics, which are valid both in the bulk and in an inhomogeneous situation. The screened potential phi arising in the formalism happens to coincide with the linearized mean-field approximation for the immersion free energy of two external unit charges. Phi is shown to decay as a repulsive f(phi)(x,x('))/y(3) interaction, whatever the density profiles may be. f(phi)(x,x(')) takes a factorized dipolar structure f(phi)(x,x('))=D(phi)(x)D(phi)(x(')) for distances x and x(') larger than the maximum of the closest approach distances b(alpha)'s to the wall for every species alpha. Moreover, we devise a reorganization of resummed diagrammatics, which is adequate for the determination of the large-distance behavior of correlations, and we prove that, when all species have the same approach distance b to the wall, f(alphaalpha('))(x,x(');b)=D(alpha)(x)D(alpha('))(x(')). In this case, the leading tail of the effective electrostatic interaction between two like charges at the same distance x from a single wall is repulsive. Results are independent of charge magnitudes, of excluded-volume sphere sizes, and of the existence of a surface charge on the wall. It holds whether charges are concentrated at sphere centers or uniformly spread over their surfaces. Comparison is made with an experiment about dilute colloids where the linearized mean-field approximation proves to be relevant. At equilibrium attraction between like charges in confined geometry might arise from purely electrostatic charge charge interactions only through correlation effects not taken into account in the latter approximation. PMID- 14525076 TI - Suppression of spiral waves and spatiotemporal chaos by generating target waves in excitable media. AB - A method for suppressing spiral waves and spatiotemporal chaos in excitable media is proposed. Applying suitable periodic force to a single point, we can successfully suppress spiral waves as well as spatiotemporal chaos by generating target waves. After we turn off the external force, target waves finally disappear and the whole system which was in the state of spiral wave or spatiotemporal chaos goes to the spatially homogeneous steady state. It is shown that our control method is not sensitively model dependent. It works for a model for catalytic CO oxidation on platinum as well as for a model for cardiac muscle. PMID- 14525077 TI - Open boundaries in a cellular automata model for synchronized flow: effects of nonmonotonicity. AB - In this paper, we have discussed the traffic situations arising from the open boundary conditions (OBC) of a cellular automata model that can reproduce the synchronized flow. The model is different from the slow-to-start (STS) model in that the upper branch of the fundamental diagram in the periodic boundary conditions (PBC) is not monotonous but has an extremum. The phase diagram and the fundamental diagram of the model in the OBC are investigated. The results are compared with those of the STS model and those in the PBC. The current in the OBC as well as the density profiles in the different phases is also investigated. PMID- 14525078 TI - Unified model and reverse recovery nonlinearities of the driven diode resonator. AB - We study the origins of period doubling and chaos in the driven series resistor inductor-varactor diode (RLD) nonlinear resonant circuit. We find that resonators driven at frequencies much higher than the diode reverse recovery rate do not show period doubling. Models of chaos based on the nonlinear capacitance of the varactor diode display a reverse-recovery-like effect, and this effect strongly resembles reverse recovery of real diodes. We find for the first time that in addition to the known dependence of the reverse recovery time on past current maxima, there are also important nonlinear dependencies on pulse frequency, duty cycle, and dc voltage bias. Similar nonlinearities are present in the nonlinear capacitance models of these diodes. We conclude that a history-dependent and nonlinear reverse-recovery time is an essential ingredient for chaotic behavior of this circuit, and demonstrate for the first time that all major competing models have this effect, either explicitly or implicitly. Besides unifying the two major models of RLD chaos, our work reveals that the nonlinearities of the reverse-recovery time must be included for a complete understanding of period doubling and chaos in this circuit. PMID- 14525079 TI - Renyi entropies characterizing the shape and the extension of the phase space representation of quantum wave functions in disordered systems. AB - We discuss some properties of the generalized entropies, called Renyi entropies, and their application to the case of continuous distributions. In particular, it is shown that these measures of complexity can be divergent; however, their differences are free from these divergences, thus enabling them to be good candidates for the description of the extension and the shape of continuous distributions. We apply this formalism to the projection of wave functions onto the coherent state basis, i.e., to the Husimi representation. We also show how the localization properties of the Husimi distribution on average can be reconstructed from its marginal distributions that are calculated in position and momentum space in the case when the phase space has no structure, i.e., no classical limit can be defined. Numerical simulations on a one-dimensional disordered system corroborate our expectations. PMID- 14525080 TI - Front explosions in three-dimensional resonantly-forced oscillatory systems. AB - Interface dynamics in a three-dimensional coupled map lattice with a period-3 local map is studied. The system possesses a parameter regime where one typically finds three-phase patterns consisting of spatially uniform domains which follow the period-3 cycle and oscillate among the three different phases. The interfaces where these domains meet may exhibit complex irregular dynamics. The system also has a parameter regime of "turbulent" dynamics, which is a chaotic transient with a superexponentially long lifetime. The transition from the three-phase pattern regime to the turbulent regime is studied. As a control parameter is tuned, the interfaces between domains develop turbulent structure. The thickness of the turbulent zone remains finite up to a critical parameter value after which it is infinite. We characterize this "front explosion" transition in three-dimensional systems and compare it with the analogous transition in two-dimensional systems where the critical properties are markedly different. The front explosion in the three-dimensional resonantly-forced complex Ginzburg-Landau equation is also investigated briefly and its character differs from that in the three-dimensional coupled map lattice. PMID- 14525081 TI - Time-delay autosynchronization of the spatiotemporal dynamics in resonant tunneling diodes. AB - The double barrier resonant tunneling diode exhibits complex spatiotemporal patterns including low-dimensional chaos when operated in an active external circuit. We demonstrate how autosynchronization by time-delayed feedback control can be used to select and stabilize specific current density patterns in a noninvasive way. We compare the efficiency of different control schemes involving feedback in either local spatial or global degrees of freedom. The numerically obtained Floquet exponents are explained by analytical results from linear stability analysis. PMID- 14525082 TI - Synchronization of spatiotemporal patterns in locally coupled excitable media. AB - The synchronization of two distributed Belousov-Zhabotinsky systems is experimentally and theoretically investigated. Symmetric local coupling of the systems is made possible with the use of a video camera-projector scheme. The spatial disorder of the coupled systems, with random initial configurations of spirals, gradually decreases until a final state is attained, which corresponds to a synchronized state with a single spiral in each system. The experimental observations are confirmed with numerical simulations of two identical Oregonator models with symmetric local coupling, and a systematic study reveals generalized synchronization of spiral waves. Several different types of synchronization attractors are distinguished. PMID- 14525083 TI - Zeta function zeros, powers of primes, and quantum chaos. AB - We present a numerical study of Riemann's formula for the oscillating part of the density of the primes and their integer powers. The formula consists of an infinite series of oscillatory terms, one for each zero of the zeta function on the critical line, and was derived by Riemann in his paper on primes, assuming the Riemann hypothesis. We show that high-resolution spectral lines can be generated by the truncated series at all integer powers of primes and demonstrate explicitly that the relative line intensities are correct. We then derive a Gaussian sum rule for Riemann's formula. This is used to analyze the numerical convergence of the truncated series. The connections to quantum chaos and semiclassical physics are discussed. PMID- 14525084 TI - Pattern formation in a monolayer of magnetic spheres. AB - Pattern formation is investigated for a vertically vibrated monolayer of magnetic spheres. The spheres of diameter D encase cylindrical magnetic cores of length l. For large D/l, we find that the particles form a hexagonal-close-packed pattern in which the particles' dipole vectors assume a macroscopic circulating vortical pattern. For smaller D/l, the particles form concentric rings. The static configurational magnetic energy (which depends on D/l) appears to be a determining factor in pattern selection even though the experimental system is driven and dissipative. PMID- 14525085 TI - Isospectrality in chaotic billiards. AB - We consider a modification of isospectral cavities whereby the classical dynamics changes from pseudointegrable to chaotic. We construct an example where we can prove that isospectrality is retained. We then demonstrate this explicitly in microwave resonators. PMID- 14525086 TI - Coherent manipulation of quantum delta-kicked dynamics: faster-than-classical anomalous diffusion. AB - Large transporting regular islands are found in the classical phase space of a modified kicked rotor system in which the kicking potential is reversed after every two kicks. The corresponding quantum system, for a variety of system parameters and over long time scales, is shown to display energy absorption that is significantly faster than that associated with the underlying classical anomalous diffusion. The results are of interest to areas of both quantum chaos and quantum control. PMID- 14525087 TI - Rules of selection for spontaneous coherent states in mesoscopic systems: using the microcavity laser as an analog study. AB - The selection rules for spontaneous coherent waves in mesoscopic systems are experimentally studied using the transverse patterns of a microcavity laser and theoretically analyzed using the theory of SU(2) coherent states. Comparison of the experimental results with the theoretical analyses reveals that an amplitude factor A should be included in the representation of the partially coherent states. The determination of the amplitude factor A is found to be associated with the constraint of minimum energy uncertainty. PMID- 14525088 TI - Fractal entropy of a chain of nonlinear oscillators. AB - We study the time evolution of a chain of nonlinear oscillators. We focus on the fractal features of the spectral entropy and analyze its characteristic intermediate time scales as a function of the nonlinear coupling. A Brownian motion is recognized with an analytic power-law dependence of its diffusion coefficient on the coupling. PMID- 14525089 TI - Period locking due to delayed feedback in a laser with saturable absorber. AB - We consider laser with saturable absorber operating in the pulsating regime that is subject to delayed feedback. Alone, both the saturable absorber and delayed feedback cause the clockwise output to become unstable to periodic output via Hopf bifurcations. The delay feedback causes the laser pulse period to lock to an integer fraction of the feedback time. We derive a map from the original model to describe the periodic pulsations of the laser. Equations for the period of the laser predict the occurrence of the different locking states as well as the value of the pump when there is a switch between the locked states. PMID- 14525090 TI - Hole-defect chaos in the one-dimensional complex Ginzburg-Landau equation. AB - We study the spatiotemporally chaotic dynamics of holes and defects in the one dimensional (1D) complex Ginzburg-Landau equation (CGLE). We focus particularly on the self-disordering dynamics of holes and on the variation in defect profiles. By enforcing identical defect profiles and/or smooth plane wave backgrounds, we are able to sensitively probe the causes of the spatiotemporal chaos. We show that the coupling of the holes to a self-disordered background is the dominant mechanism. We analyze a lattice model for the 1D CGLE, incorporating this self-disordering. Despite its simplicity, we show that the model retains the essential spatiotemporally chaotic behavior of the full CGLE. PMID- 14525091 TI - Oscillatory amplification of stochastic resonance in excitable systems. AB - We study systems which combine both oscillatory and excitable properties, and hence intrinsically possess two internal frequencies, responsible for standard spiking and for small amplitude oscillatory limit cycles (Canard orbits). We show that in such a system the effect of stochastic resonance can be amplified by application of an additional high-frequency signal, which is in resonance with the oscillatory frequency. It is important that for this amplification one needs much lower noise intensities as for conventional stochastic resonance in excitable systems. PMID- 14525092 TI - Lifetimes of noisy repellors. AB - We study the effects of additive noise on the lifetimes of chaotic repellors. Using first-order perturbation theory, we argue that noise will increase the lifetime if the escape holes lie in regions where the unperturbed density is higher than that in the immediate vicinity and that it decreases if the density is lower. Numerical experiments support the qualitative conclusions also beyond perturbation theory. PMID- 14525093 TI - Overlapping of nonlinear resonances and the problem of quantum chaos. AB - The motion of a nonlinearly oscillating particle under the influence of a periodic sequence of short impulses is investigated. We analyze the Schrodinger equation for the universal Hamiltonian. It is shown that the quantum criterion of overlapping of resonances is of the form lambdaK>or=1, where K is the classical coefficient of stochasticity and lambda is the functional defined with the use of Mathieu functions. The area of the maximal values of lambda is determined. The idea about the emerging of quantum chaos due to the adiabatic motion along the curves of Mathieu characteristics at multiple passages through the points of branching is advanced. PMID- 14525094 TI - Rapid convergence of time-averaged frequency in phase synchronized systems. AB - Numerical and experimental evidences are presented to show that many phase synchronized systems of nonidentical chaotic oscillators, where the chaotic state is reached through a period-doubling cascade, show rapid convergence of the time averaged frequency. The speed of convergence toward the natural frequency scales as the inverse of the measurement period. The results also suggest an explanation for why such chaotic oscillators can be phase synchronized. PMID- 14525095 TI - Boundary effects in the stepwise structure of the Lyapunov spectra for quasi-one dimensional systems. AB - Boundary effects in the stepwise structure of the Lyapunov spectra and corresponding wavelike structure of the Lyapunov vectors are discussed numerically in quasi-one-dimensional systems of many hard disks. Four different types of boundary conditions are constructed by combinations of periodic boundary conditions and hard-wall boundary conditions, and each leads to different stepwise structures of the Lyapunov spectra. We show that for some Lyapunov exponents in the step region, the spatial y component of the corresponding Lyapunov vector deltaq(yj), divided by the y component of momentum p(yj), exhibits a wavelike structure as a function of position q(xj) and time t. For the other Lyapunov exponents in the step region, the y component of the corresponding Lyapunov vector deltaq(yj) exhibits a time-independent wavelike structure as a function of q(xj). These two types of wavelike structure are used to categorize the type and sequence of steps in the Lyapunov spectra for each different type of boundary condition. PMID- 14525096 TI - Complex Ginzburg-Landau equation with nonlocal coupling. AB - A Ginzburg-Landau-type equation with nonlocal coupling is derived systematically as a reduced form of a universal class of reaction-diffusion systems near the Hopf bifurcation point and in the presence of another small parameter. The reaction-diffusion systems to be reduced are such that the chemical components constituting local oscillators are nondiffusive or hardly diffusive, so that the oscillators are almost uncoupled, while there is an extra diffusive component which introduces effective nonlocal coupling over the oscillators. Linear stability analysis of the reduced equation about the uniform oscillation is also carried out. This revealed that new types of instability which can never arise in the ordinary complex Ginzburg-Landau equation are possible, and their physical implication is briefly discussed. PMID- 14525097 TI - Synchronization law for a van der Pol array. AB - We explore the transition to in-phase synchronization in globally coupled oscillator arrays, and compare results for van der Pol arrays with Josephson junction arrays. Our approach yields in each case an analytically tractable iterative map; the resulting stability formulas are simple because the expansion procedure identifies natural parameter groups. A third example, an array of Duffing-van der Pol oscillators, is found to be of the same fundamental type as the van der Pol arrays, but the Josephson arrays are fundamentally different owing to the absence of self-resonant interactions. PMID- 14525098 TI - Signatures of dynamical tunneling in semiclassical quantum dots. AB - We study transport in large, and strongly open, quantum dots, which might typically be viewed as lying well within the semiclassical regime. The low temperature magnetoresistance of these structures exhibits regular fluctuations, with just a small number of dominant frequency components, indicative of the presence of dynamical tunneling into regular orbits. Support for these ideas is provided by the results of numerical simulations, which reveal wave function scarring by classically inaccessible orbits, which is found to persist even in the presence of a moderately disordered dot potential. Our results suggest that dynamical tunneling may play a more generic role in transport through mesoscopic structures than has thus far been appreciated. PMID- 14525099 TI - Influence of buoyancy on thermocapillary oscillations in a two-layer system. AB - The influence of buoyancy on thermocapillary oscillations was investigated. Nonlinear simulations of standing Marangoni waves in a real two-layer system of fluids were performed. Both the subcritical and supercritical oscillatory regimes were studied. It was found that buoyancy leads to the regularization and suppression of oscillations. The conditions for observation of different types of instability are discussed. PMID- 14525100 TI - Evolution of turbulence in an oscillatory flow in a smooth-walled channel: a viscous secondary instability mechanism. AB - In this paper, a comparison is made of the evolution of turbulence in oscillatory channel flows with a zero-mean velocity in two and three dimensions, using the numerical technique of the lattice Boltzmann method. The results confirm a primary two-dimensional instability. Evidence is shown of a secondary, viscous three-dimensional instability mechanism acting in the oscillatory boundary layer, which is consistent with experimental observations. PMID- 14525101 TI - Convective instability of an acidity front in Hele-Shaw cells. AB - Density fingering arising in the acid-catalyzed chlorite-tetrathionate reaction is investigated experimentally in Hele-Shaw cells of various thickness. Upward propagating chemical fronts are stable for all gapwidths examined. The initial evolution of the downward propagating planar fronts resulting in a cellular structure has been characterized by dispersion curves and it is found that the convectively unstable regime increases as the gapwidth increases. From the dispersion curves, the change in the characteristics of the most unstable mode and in the marginal wave number, separating the stable and unstable modes, is determined quantitatively. PMID- 14525102 TI - Bottleneck effect in three-dimensional turbulence simulations. AB - At numerical resolutions around 512(3) and above, three-dimensional energy spectra from turbulence simulations begin to show noticeably shallower spectra than k(-5/3) near the dissipation wave number ("bottleneck effect"). This effect is shown to be significantly weaker in one-dimensional spectra such as those obtained in wind tunnel turbulence. The difference can be understood in terms of the transformation between the one-dimensional and three-dimensional energy spectra under the assumption that the turbulent velocity field is isotropic. Transversal and longitudinal energy spectra are similar and can both accurately be computed from the full three-dimensional spectra. Second-order structure functions are less susceptible to the bottleneck effect and may be better suited for inferring the scaling exponent from numerical simulation data. PMID- 14525103 TI - Strongly nonlinear long gravity waves in uniform shear flows. AB - Long surface gravity waves of finite amplitude in uniform shear flows are considered by using an asymptotic model derived under the assumption that the aspect ratio between wavelength and water depth is small. Since its derivation requires no assumption on wave amplitude, the model can be used to describe arbitrary amplitude waves. It is shown that the simple model captures the interesting features of strongly nonlinear solitary waves observed in previous numerical solutions. When compared with the case of zero vorticity, the solitary wave in uniform shear flows is wider when propagating upstream (opposite to the direction of surface drift), while it is narrower when propagating downstream. For the upstream-propagating solitary wave, a stationary recirculating eddy appears at the bottom when wave amplitude exceeds the critical value. For the case of downstream propagation, no eddy forms at the bottom but the solitary wave becomes more peaked, yielding a cusp at the critical wave amplitude, beyond which the solitary wave has a round wave profile. Although the appearance of the derivative singularity is inconsistent with the long-wave assumption in the model, round wave profiles away from the singularity are qualitatively similar to numerical solutions and observation. PMID- 14525104 TI - Supercritical vaporization: distinguishable fluid regions. AB - The vaporization of a cold fluid pocket suddenly set into a hot, otherwise quiescent, atmosphere at supercritical pressures is analyzed. It is shown that at supercritical but moderate pressures and high ambient temperatures, the position where the thermal expansion coefficient reaches its maximum defines a relatively thin transition region that separates two distinguishable fluid regions: A cold liquidlike region where the heat transport is dominated by conduction, and a hot gaslike region where the heat transport is convective and conductive owed to the thermal expansion induced Stefan flow. PMID- 14525105 TI - Intermittency of velocity fluctuations in turbulent thermal convection. AB - We analyze velocity fluctuations in turbulent Rayleigh-Benard convection. The velocity measurements were taken at the center of an aspect-ratio-one convection cell filled with water. The measured probability density functions of the velocity difference over a time interval tau are found to change with tau, indicating that the velocity fluctuations are intermittent. The velocity intermittency can be well characterized by the She-Leveque hierarchical structure. Our analyses further show that the vertical velocity component has distinct statistical features from the horizontal components. This result indicates that the vertical direction is special and buoyancy is important even at the center of the convection cell. PMID- 14525106 TI - Systematic weakly nonlinear analysis of radial viscous fingering. AB - We present a weakly nonlinear analysis of the interface dynamics in a radial Hele Shaw cell driven by both injection and rotation. We extend the systematic expansion introduced in [E. Alvarez-Lacalle et al., Phys. Rev. E 64, 016302 (2001)] to the radial geometry, and compute explicitly the first nonlinear contributions. We also find the necessary and sufficient condition for the uniform convergence of the nonlinear expansion. Within this region of convergence, the analytical predictions at low orders are compared satisfactorily to exact solutions and numerical integration of the problem. This is particularly remarkable in configurations (with no counterpart in the channel geometry) for which the interplay between injection and rotation allows that condition to be satisfied at all times. In the case of the purely centrifugal forcing we demonstrate that nonlinear couplings make the interface more unstable for lower viscosity contrast between the fluids. PMID- 14525107 TI - Similarity scaling of pressure fluctuation in turbulence. AB - Pressure fluctuation was measured in a turbulent jet using a condenser microphone and piezoresistive transducer. The power-law exponent and proportional constant of normalized pressure spectrum are discussed from the standpoint of Kolmogorov universal scaling. The clear power law with scaling exponent close to -7/3 was confirmed in the range of 6000 and mu(r)>0), arrive causally before the negative index effects germinate completely. Comparisons with approximate analytical results demonstrate the presence of the negative index of refraction properties in the continuous wave portion of the signals. This dynamic pulse reshaping between the positive and negative index of refraction wave components causes an apparent delay in the realization of the negative index of refraction properties. Pulse broadening of the signal tails is associated with both dispersion and a larger negative index of refraction seen by the associated wave components. PMID- 14525142 TI - Simplified thermal lattice Boltzmann model for incompressible thermal flows. AB - Considering the fact that the compression work done by the pressure and the viscous heat dissipation can be neglected for the incompressible flow, and its relationship with the gradient term in the evolution equation for the temperature in the thermal energy distribution model, a simplified thermal energy distribution model is proposed. This thermal model does not have any gradient term and is much easier to be implemented. This model is validated by the numerical simulation of the natural convection in a square cavity at a wide range of Rayleigh numbers. Numerical experiments showed that the simplified thermal model can keep the same order of accuracy as the thermal energy distribution model, but it requires much less computational effort. PMID- 14525143 TI - Directed geometrical worm algorithm applied to the quantum rotor model. AB - We discuss the implementation of a directed geometrical worm algorithm for the study of quantum link-current models. In this algorithm the Monte Carlo updates are made through the biased reptation of a worm through the lattice. A directed algorithm is an algorithm where, during the construction of the worm, the probability for erasing the immediately preceding part of the worm, when adding a new part, is minimal. We introduce a simple numerical procedure for minimizing this probability. The procedure only depends on appropriately defined local probabilities and should be generally applicable. Furthermore, we show how correlation functions C(r,tau) can be straightforwardly obtained from the probability of a worm to reach a site (r,tau) away from its starting point independent of whether or not a directed version of the algorithm is used. Detailed analytical proofs of the validity of the Monte Carlo algorithms are presented for both the directed and undirected geometrical worm algorithms. Results for autocorrelation times and Green's functions are presented for the quantum rotor model. PMID- 14525144 TI - Two-phase flow through fractured porous media. AB - Two-phase flow in fractured porous media is investigated by means of a direct and complete numerical solution of the generalized Darcy equations in a three dimensional discrete fracture description. The numerical model applies to arbitrary fracture network geometry, and to arbitrary distributions of permeabilities in the porous matrix and in the fractures. It is used here in order to obtain the steady-state macroscopic relative permeabilities of random fractured media. Results are presented as functions of the mean saturation and are discussed in comparison with simple models. PMID- 14525145 TI - Large full band gaps for photonic crystals in two dimensions computed by an inverse method with multigrid acceleration. AB - In this study, two fast and accurate methods of inverse iteration with multigrid acceleration are developed to compute band structures of photonic crystals of general shape. In particular, we report two-dimensional photonic crystals of silicon air with an optimal full band gap of gap-midgap ratio Deltaomega/omega(mid)=0.2421, which is 30% larger than ever reported in the literature. The crystals consist of a hexagonal array of circular columns, each connected to its nearest neighbors by slender rectangular rods. A systematic study with respect to the geometric parameters of the photonic crystals was made possible with the present method in drawing a three-dimensional band-gap diagram with reasonable computing time. PMID- 14525146 TI - Efficient algorithm on a nonstaggered mesh for simulating Rayleigh-Benard convection in a box. AB - An efficient semi-implicit second-order-accurate finite-difference method is described for studying incompressible Rayleigh-Benard convection in a box, with sidewalls that are periodic, thermally insulated, or thermally conducting. Operator-splitting and a projection method reduce the algorithm at each time step to the solution of four Helmholtz equations and one Poisson equation, and these are solved by fast direct methods. The method is numerically stable even though all field values are placed on a single nonstaggered mesh commensurate with the boundaries. The efficiency and accuracy of the method are characterized for several representative convection problems. PMID- 14525147 TI - Phase transition in the Ising model on a small-world network with distance dependent interactions. AB - We study the collective behavior of an Ising system on a small-world network with the interaction J(r) proportional to r(-alpha), where r represents the Euclidean distance between two nodes. In the case of alpha=0 corresponding to the uniform interaction, the system is known to possess a phase transition of the mean-field nature, while the system with the short-range interaction (alpha-->infinity) does not exhibit long-range order at any finite temperature. The Monte Carlo simulations are performed at various values of alpha, and the critical value alpha(c) beyond which the long-range order does not emerge is estimated to be 0. Thus, concluded is the absence of a phase transition in the system with the algebraically decaying interaction r(-alpha) for any nonzero positive value of alpha. PMID- 14525148 TI - Exact solution of a generalized model for surface deposition. AB - We consider a model for surface deposition in one dimension, in the presence of both precursor-layer diffusion and desorption. The model is a generalization that includes random sequential adsorption (RSA), accelerated RSA, and growth-and coalescence models as special cases. Exact solutions are obtained for the model for both its lattice and continuum versions. Expressions are obtained for physically important quantities such as the surface coverage, average island size, mass-adsorption efficiency, and the process efficiency. The connection between a limiting case of the model and epidemic models is discussed. PMID- 14525149 TI - Pecularities of Bethe-like approximations and long-range-interaction Ising models. AB - The mean-field approximation and the Bethe approximation are two of the most often used approximations when one wants to obtain approximations of the phase diagrams and the critical temperature of lattice spin systems. Both can and have often been generalized to produce what are known as cluster mean-field and Bethe approximations. Generally, three characteristics are associated with these approximations. First, they give upper bounds to the critical temperature; second, considering larger clusters will result in better approximations; and third, the Bethe approximation is better than the corresponding mean-field approximation. We show what we believe to be a rather surprising result that, for one-dimensional Ising models with algebraically decaying interactions falling off slowly enough, the Bethe cluster approximations violate all three of these characteristics. PMID- 14525150 TI - Diffusion-limited friendship network: a model for six degrees of separation. AB - A dynamic model of a society is studied where each person is an uncorrelated and noninteracting random walker. A dynamical random graph represents the acquaintance network of the society whose nodes are the individuals and links are the pairs of mutual friendships. This network exhibits a different percolationlike phase transition in all dimensions. On introducing simultaneous death and birth rates in the population, we show that the friendship network shows the six degrees of separation for ever after where the precise value of the network diameter depends on the death/birth rate. A susceptible-infected susceptible-type model of disease spreading shows that this society always remains healthy if the population density is less than certain threshold value. PMID- 14525151 TI - Mean escape time over a fluctuating barrier. AB - An approximate method for studying activation over a fluctuating barrier of potential is proposed. It involves considering separately the slow and fast components of barrier fluctuations, and it applies for any value of their correlation time tau. It gives exact results for the limiting values tau-->0 and tau--> infinity, and the agreement with numerics in between is also excellent, both for dichotomic and Gaussian barrier perturbations. PMID- 14525152 TI - Piecewise linear emulation of propagating fronts as a method for determining their speeds. AB - We use piecewise linear terms to emulate the polynomial nonlinear terms in a typical reaction-diffusion equation, replacing it thus with a set of simple linear inhomogeneous differential equations. The resulting analytic solution constitutes an excellent approximation to the exact propagating front, as is explicitly shown in the case of cubic and quintic nonlinearities, yielding also the correct value for the physically selected speed of the observable front. Such a piecewise linear emulation can be used for any nonlinearity, giving therefore a very reliable and accurate method for determining the selected speed of fronts invading unstable states, especially pushed fronts. PMID- 14525153 TI - Phase separation in coupled chaotic maps on fractal networks. AB - The phase ordering dynamics of coupled chaotic maps on fractal networks is investigated. The statistical properties of the systems are characterized by means of the persistence probability of equivalent spin variables that define the phases. The persistence saturates and phase domains freeze for all values of the coupling parameter as a consequence of the fractal structure of the networks, in contrast to the phase transition behavior previously observed in regular Euclidean lattices. Several discontinuities and other features found in the saturation persistence curve as a function of the coupling are explained in terms of changes of stability of local phase configurations on the fractals. PMID- 14525154 TI - Oscillating bubbles at the tips of optical fibers in liquid nitrogen. AB - We report that a bubble with a radius of a few micrometers may be created at a precise location on a metal-coated optical fiber tip immersed in liquid nitrogen by microsecond optical pulses with peak powers of less than 20 mW. Dynamic optical measurements reveal that after termination of the optical pulse the bubble exhibits stable oscillations for several tens of microseconds, at frequencies up to several megahertz, as it slowly collapses. PMID- 14525155 TI - Generation of high-order harmonics in plasmas of multicharged atomic ions produced by an intense laser pulse. AB - The yield of high-order harmonics has been derived for relativistic plasmas of multicharged atomic ions produced by an intense, linearly polarized laser pulse. Harmonics of the laser field are excited at the elastic electron-ion collisions in plasmas. In the nonrelativistic case only odd harmonics can be excited. The ultrarelativistic limit is considered when both even and odd harmonics are excited with the comparable efficiencies. Drift electron velocities are assumed to be small compared to the quiver electron velocities. It is shown that the harmonic yields do not depend on the instantaneous energy distribution of electrons produced at the multiple ionization on the leading edge of the laser pulse. The harmonic conductivity is a tensor with odd components along the laser polarization and with even components along the direction of propagation of the laser pulse. This conductivity tensor has been derived analytically in the ultrarelativistic limit. Possible observation of even harmonics is discussed with respect to the Weibel effect in laser plasmas. PMID- 14525156 TI - Jeans instability in partially ionized self-gravitating dusty plasmas. AB - By employing the Boltzmann distributions for electrons and ions and by retaining the full dynamics of charged dust and neutral fluids, we derive a dispersion law for coupled dust-acoustic and neutral sound waves in partially ionized self gravitating dusty plasmas. This dispersion law exhibits new classes of Jeans instability in both collisionless and highly collisional regimes. The result should help understand the origin of molecular cloud collapse in interstellar space. PMID- 14525157 TI - Dust charge fluctuations in a magnetized dusty plasma. AB - Charging currents of electrons and ions to a spherical dust grain in a uniform magnetized dusty plasma have been examined. It is found that the external magnetic field reduces the charging currents, thereby decreasing the dust charge fluctuation damping of a low-frequency electrostatic wave in a dusty plasma. PMID- 14525158 TI - Numerical method for integrodifferential generalized Langevin and master equations. AB - We show that the integrodifferential generalized Langevin and non-Markovian master equations can be transformed into larger sets of ordinary-differential equations. On the basis of this transformation we develop a numerical method for solving such integrodifferential equations. Physically motivated example calculations are performed to demonstrate the accuracy and convergence of the method. PMID- 14525159 TI - Wavelet treatment of radial distribution functions of solutes. AB - Discrete wavelets are applied to parametrize the radial distribution functions of hydrated ions and hydrophobic solutes. The data on radial distribution functions are derived from the integral equation theory and neutron scattering experiment. The Coifman and the discrete Meyer basis sets are used for the wavelet approximation. The quality of the approximation is verified by calculations of the solvation energy, the coordination number, and the change in chemical potential of solutes. PMID- 14525160 TI - Comment on "Analog of Planck's formula and effective temperature in classical statistical mechanics far from equilibrium". AB - The attempt of Carati and Galgani [Phys. Rev. E 61, 4791 (2000)] to derive the Planck formula as though it were due to long relaxation times is shown to be correct only if there is a single temperature and therefore no transients. PMID- 14525162 TI - Short-distance correlation properties of the Lieb-Liniger system and momentum distributions of trapped one-dimensional atomic gases. AB - We derive exact closed-form expressions for the first few terms of the short distance Taylor expansion of the one-body correlation function of the Lieb Liniger gas. As an intermediate result, we obtain the high-p asymptotics of the momentum distribution of both free and harmonically trapped atoms and show that it obeys a universal 1/p(4) law for all values of the interaction strength. We discuss the ways to observe the predicted momentum distributions experimentally, regarding them as a sensitive identifier for the Tonks-Girardeau regime of strong correlations. PMID- 14525163 TI - Controlling spin exchange interactions of ultracold atoms in optical lattices. AB - We describe a general technique that allows one to induce and control strong interaction between spin states of neighboring atoms in an optical lattice. We show that the properties of spin exchange interactions, such as magnitude, sign, and anisotropy, can be designed by adjusting the optical potentials. We illustrate how this technique can be used to efficiently "engineer" quantum spin systems with desired properties, for specific examples ranging from scalable quantum computation to probing a model with complex topological order that supports exotic anyonic excitations. PMID- 14525164 TI - Experimental observation of a superfluid gyroscope in a dilute Bose-Einstein condensate. AB - We have observed a three-dimensional gyroscopic effect associated with a vortex in a dilute Bose-Einstein condensed gas. A condensate with a vortex possesses a single quantum of circulation, and this causes the plane of oscillation of the scissors mode to precess around the vortex line. We have measured the precession rate of the scissors oscillation. From this we deduced the angular momentum associated with the vortex line and found a value close to Planck's over 2pi per particle, as predicted for a superfluid. PMID- 14525165 TI - Berry phase for a spin 1/2 particle in a classical fluctuating field. AB - The effect of fluctuations in the classical control parameters on the Berry phase of a spin 1/2 interacting with an adiabatically cyclically varying magnetic field is analyzed. It is explicitly shown that in the adiabatic limit dephasing is due to fluctuations of the dynamical phase. PMID- 14525166 TI - Mixed state geometric phases, entangled systems, and local unitary transformations. AB - The geometric phase for a pure quantal state undergoing an arbitrary evolution is a "memory" of the geometry of the path in the projective Hilbert space of the system. We find that Uhlmann's geometric phase for a mixed quantal state undergoing unitary evolution depends not only on the geometry of the path of the system alone but also on a constrained bilocal unitary evolution of the purified entangled state. We analyze this in general, illustrate it for the qubit case, and propose an experiment to test this effect. We also show that the mixed state geometric phase proposed recently in the context of interferometry requires unilocal transformations and is therefore essentially a property of the system alone. PMID- 14525167 TI - Quantum state tomography of dissociating molecules. AB - Using tomographic reconstruction we determine the complete internuclear quantum state, represented by the Wigner function, of a dissociating I2 molecule based on femtosecond time resolved position and momentum distributions of the atomic fragments. The experimental data are recorded by timed ionization of the photofragments with an intense 20 fs laser pulse. Our reconstruction method, which relies on Jaynes's maximum entropy principle, will also be applicable to time resolved position or momentum data obtained with other experimental techniques. PMID- 14525168 TI - Dissipative flow and vortex shedding in the Painleve boundary layer of a Bose Einstein condensate. AB - This paper addresses the drag force and formation of vortices in the boundary layer of a Bose-Einstein condensate stirred by a laser beam following the experiments of Phys. Rev. Lett. 83, 2502 (1999)]. We make our analysis in the frame moving at constant speed where the beam is fixed. We find that there is always a drag around the laser beam. We also analyze the mechanism of vortex nucleation. At low velocity, there are no vortices and the drag has its origin in a wakelike phenomenon: This is a particularity of trapped systems since the density gets small in an extended region. The shedding of vortices starts only at a threshold velocity and is responsible for a large increase in drag. This critical velocity for vortex nucleation is lower than the critical velocity computed for the corresponding 2D problem at the center of the cloud. PMID- 14525169 TI - Wave nature of biomolecules and fluorofullerenes. AB - We demonstrate quantum interference for tetraphenylporphyrin, the first biomolecule exhibiting wave nature, and for the fluorofullerene C60F48 using a near-field Talbot-Lau interferometer. For the porphyrins, which are distinguished by their low symmetry and their abundant occurrence in organic systems, we find the theoretically expected maximal interference contrast and its expected dependence on the de Broglie wavelength. For C60F48, the observed fringe visibility is below the expected value, but the high contrast still provides good evidence for the quantum character of the observed fringe pattern. The fluorofullerenes therefore set the new mark in complexity and mass (1632 amu) for de Broglie wave experiments, exceeding the previous mass record by a factor of 2. PMID- 14525170 TI - Interference study of the chi c0(13P0) in the reaction -pp-->pi0pi0. AB - Fermilab experiment E835 has observed (-)pp annihilation production of the charmonium state chi(c0) and its subsequent decay into pi(0)pi(0). Although the resonant amplitude is an order of magnitude smaller than that of the nonresonant continuum production of pi(0)pi(0), an enhanced interference signal is evident. A partial wave expansion is used to extract physics parameters. The amplitudes J=0 and 2, of comparable strength, dominate the expansion. Both are accessed by L=1 in the entrance (-)pp channel. The product of the input and output branching fractions is determined to be B((-)pp-->chi(c0))xB(chi(c0)-->pi(0)pi(0))=(5.09+/ 0.81+/-0.25)x10(-7). PMID- 14525172 TI - etapi and eta'pi spectra and interpretation of possible exotic JPC=1-+ mesons. AB - We discuss a coupled channel analysis of the etapi and eta'pi systems produced in pi(-)p interactions at 18 GeV/c. We show that known Q(-)Q resonances, together with residual soft meson-meson rescattering, saturate the spectra including the exotic J(PC)=1(-+) channel. The possibility of a narrow exotic resonance at a mass near 1.6 GeV/c(2) cannot, however, be ruled out. PMID- 14525171 TI - Beam-polarization asymmetries for the p(gamma-->, K+)Lambda and p(gamma-->, K+)Sigma(0) reactions for E(gamma)=1.5-2.4 GeV. AB - Beam polarization asymmetries for the p(gamma-->,K+)Lambda and p(gamma- >,K+)Sigma(0) reactions are measured for the first time for E(gamma)=1.5-2.4 GeV and 0.6B+X, we derive new positivity constraints on spin observables and study their implications for theoretical models in view, in particular, of accounting for future data from the polarized pp collider at Brookhaven RHIC. We find that the single transverse spin asymmetry A(N), in the central region for several processes, for example, jet production, direct photon production, and lepton-pair production, is expected to be such that |A(N)| < or approximately 1/2, rather than the usual bound |A(N)|< or =1. PMID- 14525176 TI - Elliptic flow at large transverse momenta from quark coalescence. AB - We show that hadronization via quark coalescence enhances hadron elliptic flow at large p(perpendicular) relative to that of partons at the same transverse momentum. Therefore, compared to earlier results based on covariant parton transport theory, more moderate initial parton densities dN/deta(b=0) approximately 1500-3000 can explain the differential elliptic flow v(2)(p(perpendicular)) data for Au+Au reactions at sqrt[s]=130 and 200A GeV from BNL RHIC. In addition, v(2)(p(perpendicular)) could saturate at about 50% higher values for baryons than for mesons. If strange quarks have weaker flow than light quarks, hadron v(2) at high p(perpendicular) decreases with relative strangeness content. PMID- 14525177 TI - Chronology of particle emission from the E/A=61 MeV 36Ar+27Al reaction. AB - The emission time chronology of neutrons, protons, and deuterons from the E/A=61 MeV 36Ar+27Al reaction is deduced from two-particle correlation functions. PMID- 14525178 TI - Relativistic linear response wave functions and dynamic scattering tensor for the ns1/2 states in hydrogenlike atoms. AB - We report a novel closed-form analytic representation for the linear response relativistic wave function of the hydrogenic ns(1/2) level that is exposed to dipole radiation of frequency omega. This result is derived by means of a direct analytical solution of the inhomogeneous omega-dependent Dirac equation. The utility of the formulas obtained is demonstrated by new analytic and numerical calculations of the static and dynamic relativistic dynamic polarizabilities of the lowest hydrogenic ns(1/2) states. PMID- 14525179 TI - Time-resolved double ionization with few cycle laser pulses. AB - Ionization of D2 launches a vibrational wave packet on the ground state of D+2. Removal of the second electron places a pair of D+ ions onto a Coulombic potential. Measuring the D+ kinetic energy determines the time delay between the first and the second ionization. Caught between a falling ionization and a rapidly rising intensity, the typical lifetime of the D+2 intermediate is less than 5 fs when an intense 8.6 fs laser pulse is used. We simulate Coulomb explosion imaging of the ground state wave function of D2 by a 4 fs optical pulse and compare with our experimental observations. PMID- 14525180 TI - Observation of large Kerr nonlinearity at low light intensities. AB - We report an experimental observation of large Kerr nonlinearity with vanishing linear susceptibilities in coherently prepared four-level rubidium atoms. Quantum coherence and interference manifested by electromagnetically induced transparency suppress the linear susceptibilities and greatly enhance the nonlinear susceptibilities at low light intensities. The measured Kerr nonlinearity is comparable in magnitude to the linear dispersion in a simple two-level system and is several orders of magnitude greater than the Kerr nonlinearity of a conventional three-level scheme under similar conditions. PMID- 14525181 TI - Observation of the transfer of the local angular momentum density of a multiringed light beam to an optically trapped particle. AB - We observe the spinning and orbital motion of a microscopic particle trapped within a multiringed light beam that arises from the transfer of the spin and orbital components of the light's angular momentum. The two rotation rates are measured as a function of the distance between the particle and the axis of the trapping beam. The radial dependence of these observations is found to be in close agreement with the accepted theory. PMID- 14525182 TI - Spatial correlations in the near field of random media. AB - The conventional coherence theory suggests that the fields radiated by statistically homogeneous sources correlate over spatial regions of the order of the wavelength irrespective of the distance from the surface of the source. Contrary to these predictions, we show that the spatial correlations of optical fields in close proximity of highly scattering, randomly inhomogeneous media depend on this distance and, moreover, their extent can be significantly smaller than the wavelength. The contribution of evanescent fields is experimentally demonstrated and the coherence length in the near field is shown to relate to the coherence properties at the surface which are, in turn, determined by the structural characteristics of the random media. PMID- 14525183 TI - Breakdown of wave diffusion in 2D due to loops. AB - The validity of the diffusion approximation for the intensity of multiply scattered waves is tested with numerical simulations in a strongly scattering 2D medium of finite extent. We show that the diffusion equation underestimates the intensity and attribute this to both the neglect of recurrent scattering paths and interference within diffusion theory. We present a theory to quantify this discrepancy based on counting all possible scattering paths between point scatterers. Interference phenomena, due to loop paths, are incorporated in a way similar to coherent backscattering. PMID- 14525184 TI - Photon antibunching and collective effects in the fluorescence of single bichromophoric molecules. AB - The fluorescence of individual pairs of perylenemonoimide chromophores coupled via a short rigid linker is investigated. Photon antibunching is reported, indicating collective effects in the fluorescence, which are further substantiated by the observation of collective triplet off times and triplet lifetime shortening. The experimental findings are analyzed in terms of singlet singlet and singlet-triplet annihilation based on Forster type energy transfer. The results reported here demonstrate that the statistical properties of the emission light of isolated single quantum systems can serve as a hallmark of intermolecular interactions. PMID- 14525185 TI - Spontaneously generated X-shaped light bullets. AB - We observe the formation of an intense optical wave packet fully localized in all dimensions, i.e., both longitudinally (in time) and in the transverse plane, with an extension of a few tens of fsec and microns, respectively. Our measurements show that the self-trapped wave is an X-shaped light bullet spontaneously generated from a standard laser wave packet via the nonlinear material response (i.e., second-harmonic generation), which extend the soliton concept to a new realm, where the main hump coexists with conical tails which reflect the symmetry of linear dispersion relationship. PMID- 14525186 TI - Two-beam modulation instability in noninstantaneous nonlinear media. AB - The development of modulation instability for two beams copropagating in a medium with noninstantaneous nonlinear response is investigated. The frequency dependence of the experimentally observed spectral broadening reveals the presence of two-beam coupling, resulting from the time-delayed Raman part of the third-order nonlinear susceptibility. We present an analysis of the experimental data, which yields a value of tau=27(1) fs for the lifetime of the optical phonons in silica fibers. To our knowledge, this is the first experimentally determined value for this important material parameter. PMID- 14525187 TI - Distributed delays facilitate amplitude death of coupled oscillators. AB - Coupled oscillators are shown to experience amplitude death for a much larger set of parameter values when they are connected with time delays distributed over an interval rather than concentrated at a point. Distributed delays enlarge and merge death islands in the parameter space. Furthermore, when the variance of the distribution is larger than a threshold, the death region becomes unbounded and amplitude death can occur for any average value of delay. These phenomena are observed even with a small spread of delays, for different distribution functions, and an arbitrary number of oscillators. PMID- 14525188 TI - Thermal-noise effect on the transition to Rayleigh-Benard convection. AB - We report measurements of fluctuation and roll patterns near the transition to Rayleigh-Benard convection which are consistent with a fluctuation-induced first order transition, as predicted by Swift and Hohenberg. Above onset, we find convection rolls with noise-induced fluctuations, time-dependent amplitude modulation and roll undulation, and homogeneous dislocation nucleation. PMID- 14525189 TI - Efficiency enhancement in free-electron lasers by means of concurrent rf acceleration. AB - Efficiency enhancement in free-electron lasers (FELs) using rf beam acceleration in the wiggler is described. Since the beam tube is a waveguide, there are low and high frequency resonances. Injection of low frequency power can act as an inverse-FEL accelerator concurrently with high frequency power extraction. Simulation of a FEL using this technique shows that substantial efficiency enhancements are possible without significant increases in the beam energy spread, which facilitates the use of energy recovery schemes. The technique is applicable to amplifier and oscillator configurations. PMID- 14525190 TI - Synergy of anomalous transport and radiation in the density limit. AB - The critical plasma density n(cr) above which the edge anomalous transport in tokamaks is dominated by drift resistive ballooning instability is found analytically. In this transport regime, the drastic increase of particle losses and drop of the edge temperature provoke a strong increase in impurity radiation, and thermal equilibrium does not exist if the density is ramped up above the ultimate limit n(max). Because of the nonlinear character of impurity radiation, this density limit n(max) is very close to n(cr) and practically does not change with the ion effective charge. The importance of the synergy between the anomalous transport and impurity radiation for the density limit phenomenon is confirmed by the results of numerical simulations. PMID- 14525191 TI - Observation of a structural transition for Coulomb crystals in a linear Paul trap. AB - A structural transition for laser cooled ion Coulomb crystals in a linear Paul trap just above the stability limit of parametrically resonant excitation of bulk plasma modes has been observed. In contrast to the usual spheroidal shell structures present below the stability limit, the ions arrange in a "string-of disks" configuration. The spheroidal envelopes of the string-of-disks structures are in agreement with results from cold fluid theory usually valid for ion Coulomb crystals if the ion systems are assumed to be rotating collectively. PMID- 14525192 TI - Time-dependent electron thermal flux inhibition in direct-drive laser implosions. AB - We simulate direct-drive CH target implosions with square laser pulses by a one dimensional Fokker-Planck solver combined with a hydrodynamic code, and compare the results with those simulated by the flux-limited Spitzer-Harm model. We find that the electron thermal flux inhibition is time dependent, resulting in longer density scale length, larger laser absorption, and smaller growth of Rayleigh Taylor instability. The time of peak neutron production calculated from Fokker Planck simulations agrees with experiments for both 1-ns and 400-ps pulses. PMID- 14525193 TI - Creeping friction dynamics and molecular dissipation mechanisms in glassy polymers. AB - The dissipation mechanism of nanoscale kinetic friction between an atomic force microscopy tip and a surface of amorphous glassy polystyrene has been studied as a function of two parameters: the scanning velocity and the temperature. Superposition of the friction results using the method of reduced variables revealed the dissipative behavior as an activated relaxation process with a potential barrier height of 7.0 kcal/mol, corresponding to the hindered rotation of phenyl groups around the C-C bond with the backbone. The velocity relationship with friction F(v) was found to satisfy simple fluctuation surface potential models with F proportional to const-ln(v) and F proportional to const-ln(v)2/3. PMID- 14525194 TI - Role of the entangled amorphous network in tensile deformation of semicrystalline polymers. AB - Being composed of crystalline lamellae and entangled amorphous polymeric chains in between, semicrystalline polymers always show a complicated deformation behavior under tensile deformation. In recent years, the process of tensile deformation was found to exhibit several regimes: intralamellar slipping of crystalline blocks occurs at small deformation whereas a stress-induced crystalline block disaggregation-recrystallization process occurs at a strain larger than the yield strain. The strain at this transition point is related to the interplay between the amorphous entanglement density and the stability of crystal blocks. We report experimental evidence from true stress-strain experiments that support this argument. It is emphasized that tie molecules, which connect adjacent lamellae, are of lesser importance with respect to the deformational behavior. PMID- 14525195 TI - Origins of growth stresses in amorphous semiconductor thin films. AB - Stress evolution during deposition of amorphous Si and Ge thin films is remarkably similar to that observed for polycrystalline films. Amorphous semiconductors were used as model materials to study the origins of deposition stresses in continuous films, where suppression of both strain relaxation and epitaxial strain inheritance provides considerable simplification. Our data show that bulk compression is established by surface stress, while a subsequent return to tensile stress arises from elastic coalescence processes occurring on the kinetically roughened surface. PMID- 14525196 TI - Nanowires and nanorings at the atomic level. AB - The step-flow growth mode is used to fabricate Si and Ge nanowires with a width of 3.5 nm and a thickness of one atomic layer (0.3 nm) by self-assembly. Alternating deposition of Ge and Si results in the formation of a nanowire superlattice covering the whole surface. One atomic layer of Bi terminating the surface is used to distinguish between the elements Si and Ge. A difference in apparent height is measured in scanning tunneling microscopy images for Si and Ge. Also, different kinds of two-dimensional Si/Ge nanostructures like alternating Si and Ge nanorings having a width of 5-10 nm were grown. PMID- 14525197 TI - Quantum magnetic impurities in magnetically ordered systems. AB - We discuss the problem of a spin 1/2 impurity immersed in a spin S magnetically ordered background. We show that the problem maps onto a generalization of the dissipative two level system with two independent heat baths, associated with the Goldstone modes of the magnet, that couple to different components of the impurity spin operator. Using analytical perturbative renormalization group methods and accurate numerical renormalization group we show that contrary to other dissipative models there is quantum frustration of decoherence and quasiscaling even in the strong coupling regime. We make predictions for the behavior of the impurity magnetic susceptibility. Our results may also have relevance to quantum computation. PMID- 14525198 TI - Scaling and the center-of-band anomaly in a one-dimensional Anderson model with diagonal disorder. AB - We resolve the problem of the violation of single parameter scaling at the zero energy of the Anderson tight-binding model with diagonal disorder. It follows from the symmetry properties of the tight-binding Hamiltonian that this spectral point is, in fact, a boundary between two adjacent bands. The states in the vicinity of this energy behave similarly to states at other band boundaries, which are known to violate single parameter scaling. PMID- 14525199 TI - Evidence for spontaneous spin-polarized transport in magnetic nanowires. AB - The exploitation of the spin in charge-based systems is opening revolutionary opportunities for device architecture. Surprisingly, room temperature electrical transport through magnetic nanowires is still an unresolved issue. Here, we show that ferromagnetic (Co) suspended atom chains spontaneously display an electron transport of half a conductance quantum, as expected for a fully polarized conduction channel. Similar behavior has been observed for Pd (a quasimagnetic 4d metal) and Pt (a nonmagnetic 5d metal). These results suggest that the nanowire low dimensionality reinforces or induces magnetic behavior, lifting off spin degeneracy even at room temperature and zero external magnetic field. PMID- 14525200 TI - Evidence of the key role of metal-molecule bonding in metal-molecule-metal transport experiments. AB - The transport properties of two oligothiophene derivatives, that differ only by the chemical group coupling to gold, are compared. It is shown that the role of the coupling group in the transport properties can be decoupled from that of the conjugated body of the molecules and that Se is a better electronic coupling group than S. These results are accounted for semiquantitatively within the frame of the scattering theory of transport, using results from ultraviolet photoemission spectroscopy experiments as inputs for the position in energy of the molecular orbitals with respect to the Fermi level of the electrodes. PMID- 14525202 TI - Amorphous vortex glass phase in strongly disordered superconductors. AB - We introduce a model describing vortices in strongly disordered three-dimensional superconductors. The model focuses on the topological defects, i.e., dislocation lines, in an elastic description of the vortex lattice. The model is studied using Monte Carlo simulations, revealing a glass phase at low temperatures, separated by a continuous phase transition to the high temperature resistive vortex liquid phase. The critical exponents nu approximately 1.3 and eta approximately -0.4 characterizing the transition are obtained from finite size scaling. PMID- 14525201 TI - Observation of microwave-induced zero-conductance state in Corbino rings of a two dimensional electron system. AB - Using Corbino samples we have observed oscillatory dc conductance in a high mobility two-dimensional electron system when it is subjected to crossed microwave and magnetic fields. At the strongest of the oscillation minima the conductance is found to be vanishingly small, indicating a macroscopic insulating state associated with this minimum. With increasing voltage bias, a crossover from Ohmic to electron-heating regime is observed. PMID- 14525203 TI - Vortex lattice structural transitions: a Ginzburg-Landau model approach. AB - We analyze the rhombic to square vortex lattice phase transition in anisotropic superconductors using a variant of Ginzburg-Landau theory. The mean-field phase diagram is determined to second order in the anisotropy parameter, and shows a reorientation transition of the square vortex lattice with respect to the crystal lattice. We then derive the long-wavelength elastic moduli of the lattices, and use them to show that thermal fluctuations produce a reentrant rhombic to square lattice transition line, similar to recent studies which used a nonlocal London model. PMID- 14525204 TI - Electronic model for CoO2 layer based systems: chiral resonating valence bond metal and superconductivity. AB - Takada et al. have reported superconductivity in layered Na(x)CoO(2)yH(2)O (T(c) approximately equal to 5 K). We model a reference neutral CoO2 layer as an orbitally nondegenerate spin-1/2 antiferromagnetic Mott insulator on a triangular lattice and Na(x)CoO(2)yH(2)O as electron doped Mott insulators described by a t J model. It is suggested that at optimal doping chiral spin fluctuations enhanced by the dopant dynamics lead to a gapful d-wave superconducting state. A chiral resonating valence bond (RVB) metal, a parity and time (PT) reversal violating state with condensed RVB gauge fields, with a possible weak ferromagnetism, and low temperature p-wave superconductivity are also suggested at higher dopings. PMID- 14525205 TI - Temperature chaos, rejuvenation, and memory in Migdal-Kadanoff spin glasses. AB - We use simulations within the Migdal-Kadanoff approach to probe the scales relevant for rejuvenation and memory in Ising spin glasses. First we investigate scaling laws for domain wall free energies and extract the chaos overlap length l(T,T'). Then we perform out of equilibrium simulations that follow experimental protocols. We find that (1) a rejuvenation signal arises at a length scale significantly smaller than l(T,T'), and (2) memory survives even if equilibration goes out to length scales larger than l(T,T'). Theoretical justifications of these phenomena are then considered. PMID- 14525206 TI - Effective Hamiltonian for Ga1-x MnxAs in the dilute limit. AB - We derive an effective Hamiltonian for Ga(1-x)Mn(x)As in the dilute limit, where Ga(1-x)Mn(x)As can be described in terms of spin F=3/2 polarons hopping between the Mn sites and coupled to the local Mn spins. We determine the parameters of our model from microscopic calculations. Our approach treats the large Coulomb interaction in a nonperturbative way, captures the effects of spin-orbit coupling and disorder, and is appropriate for other p-doped magnetic semiconductors. PMID- 14525207 TI - Magnetic tuning of biquadratic exchange coupling in magnetic thin films. AB - The effective interlayer coupling between antiferromagnetically coupled hard and soft ferromagnetic thin films is investigated as a function of the magnetic bit length in the hard layer, which is controlled using a magnetic recording system. The interlayer coupling is explored by studying the magnetization reversal of the soft layer. As the bit length decreases, the coupling evolves from antiferromagnetic to biquadratic to uncoupled. These results are reproduced using a micromagnetic model and determine the applicability range of Slonczewski's fluctuation model of biquadratic coupling. PMID- 14525208 TI - Interplay of collective excitations in quantum-well intersubband resonances. AB - Intersubband resonances in a semiconductor quantum well (QW) display fascinating features involving various collective excitations such as Fermi-edge singularity (FES) and intersubband plasmon (ISP). Using a density matrix approach, we treated many-body effects such as depolarization, vertex correction, and self-energy consistently for a two-subband system. We found a systematic change in resonance spectra from FES- to ISP-dominated features, as QW width or electron density is varied. Such an interplay between FES and ISP significantly changes both line shape and peak position of the absorption spectrum. We found that a cancellation of FES and ISP undresses the resonant responses and recovers the single-particle features of absorption for semiconductors with a strong nonparabolicity such as InAs, leading to a dramatic broadening of the absorption spectrum. PMID- 14525209 TI - Tomographic quantum cryptography: equivalence of quantum and classical key distillation. AB - The security of a cryptographic key that is generated by communication through a noisy quantum channel relies on the ability to distill a shorter secure key sequence from a longer insecure one. For an important class of protocols, which exploit tomographically complete measurements on entangled pairs of any dimension, we show that the noise threshold for classical advantage distillation is identical with the threshold for quantum entanglement distillation. As a consequence, the two distillation procedures are equivalent: neither offers a security advantage over the other. PMID- 14525210 TI - Entanglement of indistinguishable particles shared between two parties. AB - Using an operational definition we quantify the entanglement, E(P), between two parties who share an arbitrary pure state of N indistinguishable particles. We show that E(P)< or =E(M), where E(M) is the bipartite entanglement calculated from the mode-occupation representation. Unlike E(M), E(P) is superadditive. For example, E(P)=0 for any single-particle state, but the state |1>|1>, where both modes are split between the two parties, has E(P)=1/2. We discuss how this relates to quantum correlations between particles, for both fermions and bosons. PMID- 14525211 TI - Entanglement constrained by superselection rules. AB - Bipartite entanglement may be reduced if there are restrictions on allowed local operations. We introduce the concept of a generalized superselection rule to describe such restrictions, and quantify the entanglement constrained by it. We show that ensemble quantum information processing, where elements in the ensemble are not individually addressable, is subject to the superselection rule associated with the symmetric group (the group of permutations of elements). We prove that even for an ensemble comprising many pairs of qubits, each pair described by a pure Bell state, the entanglement per element constrained by this superselection rule goes to zero for a large number of elements. PMID- 14525212 TI - Dressed qubits. AB - Inherent gate errors can arise in quantum computation when the actual system Hamiltonian or Hilbert space deviates from the desired one. Two important examples we address are spin-coupled quantum dots in the presence of spin-orbit perturbations to the Heisenberg exchange interaction, and off-resonant transitions of a qubit embedded in a multilevel Hilbert space. We propose a "dressed qubit" transformation for dealing with such inherent errors. Unlike quantum error correction, the dressed qubit method does not require additional operations or encoding redundancy, is insensitive to error magnitude, and imposes no new experimental constraints. PMID- 14525213 TI - Measurement induced entanglement and quantum computation with atoms in optical cavities. AB - We propose a method to prepare entangled states and implement quantum computation with atoms in optical cavities. The internal states of the atoms are entangled by a measurement of the phase of light transmitted through the cavity. By repeated measurements an entangled state is created with certainty, and this entanglement can be used to implement gates on qubits which are stored in different internal degrees of freedom of the atoms. This method, based on measurement induced dynamics, has a higher fidelity than schemes making use of controlled unitary dynamics. PMID- 14525214 TI - Continuous monitoring of Rabi oscillations in a Josephson flux qubit. AB - Under resonant irradiation, a quantum system can undergo coherent (Rabi) oscillations in time. We report evidence for such oscillations in a continuously observed three-Josephson-junction flux qubit, coupled to a high-quality tank circuit tuned to the Rabi frequency. In addition to simplicity, this method of Rabi spectroscopy enabled a long coherence time of about 2.5 micros, corresponding to an effective qubit quality factor approximately 7000. PMID- 14525215 TI - Deformation and collapse of microtubules on the nanometer scale. AB - We probe the local mechanical properties of microtubules at the nanometer scale by radial indentation with a scanning force microscope tip. We find a linear elastic regime that can be described by both thin-shell theory and finite element methods, in which microtubules are modeled as hollow tubes. We also find a nonlinear regime and catastrophic collapse of the microtubules under large loads. The main physics of protein shells at the nanometer scale shows simultaneously aspects of continuum elasticity in their linear response, as well as molecular graininess in their nonlinear behavior. PMID- 14525216 TI - Cholesterol-induced modulated phase in phospholipid membranes. AB - We report the observation of a cholesterol-induced modulated phase (Pbeta) in dipalmitoyl phosphatidylcholine bilayers. It occurs below the main transition of the lipid at cholesterol concentrations of around 15 to 20 mol % and is distinct from the ripple (Pbeta') phase found in between the main and pretransitions at lower cholesterol concentrations. An electron density map of this phase, constructed from x-ray diffraction data from oriented multilayers, shows that the bilayers in this phase have a one-dimensional periodic height modulation with an amplitude of about 2.5 A. A partial phase diagram of the system deduced from diffraction data is in broad agreement with earlier studies. PMID- 14525217 TI - Shock waves in high-energy materials: the initial chemical events in nitramine RDX. AB - We extend the reactive force field ReaxFF to describe the high energy nitramine RDX and use it with molecular dynamics (MD) to study its shock-induced chemistry. We studied shock propagation via nonequilibrium MD simulations at various collision velocities. We find that for high impact velocities (>6 km/s) the RDX molecules decompose and react to form a variety of small molecules in very short time scales (<3 ps). These products are consistent with those found experimentally at longer times. For lower velocities only NO2 is formed, also in agreement with experiments. PMID- 14525218 TI - Mean-field and anomalous behavior on a small-world network. AB - We consider various equilibrium statistical mechanics models with combined short- and long-range interactions and identify the crossover to mean-field behavior, finding anomalous scaling in the width of the mean-field region, as well as in the mean-field amplitudes. We then show that this model enables us, in many cases, to determine the universal critical properties of systems on a small-world network. Finally, we consider nonequilibrium processes. PMID- 14525223 TI - Unbiased global optimization of Lennard-Jones clusters for N < or =201 using the conformational space annealing method. AB - We apply the conformational space annealing method to the Lennard-Jones clusters and find all known lowest energy configurations up to 201 atoms, without using extra information of the problem such as the structures of the known global energy minima. In addition, the robustness of the algorithm with respect to the randomness of initial conditions of the problem is demonstrated by ten successful independent runs up to 183 atoms. Our results indicate that this method is a general and yet efficient global optimization algorithm applicable to many systems. PMID- 14525219 TI - Energies of strained surfaces and barrierless formation of strained islands. PMID- 14525221 TI - Comment on "Left-handed materials do not make a perfect lens". PMID- 14525224 TI - Spin coupling between cold atoms and the thermal fluctuations of a metal surface. AB - We describe an experiment in which Bose-Einstein condensates and cold atom clouds are held by a microscopic magnetic trap near a room-temperature metal wire 500 microm in diameter. The lifetime for atoms to remain in the microtrap is measured over a range of distances down to 27 microm from the surface of the metal. We observe the loss of atoms from the microtrap due to spin flips. These are induced by radio-frequency thermal fluctuations of the magnetic field near the surface, as predicted but not previously observed. PMID- 14525225 TI - Intensity dependence of photoassociation in a quantum degenerate atomic gas. AB - We have measured the intensity dependent rate and frequency shift of a photoassociation transition in a quantum degenerate gas of 7Li. The rate increases linearly with photoassociation laser intensity for low intensities, whereas saturation is observed at higher intensities. The measured rates and shifts agree reasonably well with theory within the estimated systematic uncertainties. Several theoretically predicted saturation mechanisms are discussed, but a theory in which saturation arises because of quantum mechanical unitarity agrees well with the data. PMID- 14525226 TI - Atomic Bose and Anderson glasses in optical lattices. AB - An ultracold atomic Bose gas in an optical lattice is shown to provide an ideal system for the controlled analysis of disordered Bose lattice gases. This goal may be easily achieved under the current experimental conditions by introducing a pseudorandom potential created by a second additional lattice or, alternatively, by placing a speckle pattern on the main lattice. We show that, for a noncommensurable filling factor, in the strong-interaction limit, a controlled growing of the disorder drives a dynamical transition from superfluid to Bose glass phase. Similarly, in the weak interaction limit, a dynamical transition from superfluid to Anderson-glass phase may be observed. In both regimes, we show that even very low-intensity disorder-inducing lasers cause large modifications of the superfluid fraction of the system. PMID- 14525227 TI - Experimental creation of a fully inseparable tripartite continuous-variable state. AB - A continuous-variable tripartite entangled state is experimentally generated by combining three independent squeezed vacuum states, and the variances of its relative positions and total momentum are measured. We show that the measured values violate the separability criteria based on the sum of these quantities and prove the full inseparability of the generated state. PMID- 14525228 TI - Modulational instability and complex dynamics of confined matter-wave solitons. AB - We study the formation of bright solitons in a Bose-Einstein condensate of 7Li atoms induced by a sudden change in the sign of the scattering length from positive to negative, as reported in a recent experiment [Nature (London) 417, 150 (2002)]]. The numerical simulations are performed by using the Gross Pitaevskii equation with a dissipative three-body term. We show that a number of bright solitons is produced and this can be interpreted in terms of the modulational instability of the time-dependent macroscopic wave function of the Bose condensate. In particular, we derive a simple formula for the number of solitons that is in good agreement with the numerical results. We find that during the motion of the soliton train in an axial harmonic potential the number of solitonic peaks changes in time and the density of individual peaks shows an intermittent behavior. PMID- 14525229 TI - Conversion of an atomic Fermi gas to a long-lived molecular Bose gas. AB - We have converted an ultracold Fermi gas of 6Li atoms into an ultracold gas of 6Li2 molecules by adiabatic passage through a Feshbach resonance. Approximately 1.5 x 10(5) molecules in the least-bound, v=38, vibrational level of the X1Sigma(+)(g) singlet state are produced with an efficiency of 50%. The molecules remain confined in an optical trap for times of up to 1 s before we dissociate them by a reverse adiabatic sweep. PMID- 14525230 TI - Diffusion of asymmetric swimmers. AB - Particles moving along curved trajectories will diffuse if the curvature fluctuates sufficiently in either magnitude or orientation. We consider particles moving at a constant speed with either a fixed or a Gaussian distributed magnitude of curvature. At small speeds the diffusivity is independent of the speed. At larger particle speeds, the diffusivity depends on the speed through a novel exponent. We apply our results to intracellular transport of vesicles. In sharp contrast to thermal diffusion, the effective diffusivity increases with vesicle size and so may provide an effective means of intracellular transport. PMID- 14525231 TI - Nonperturbative flow equations from running expectation values. AB - We show that Wegner's flow equations, as recently discussed in the Lipkin model, can be solved self-consistently. This leads to a nonlinear differential equation which fully determines the order parameter as a function of the dimensionless coupling constant, even across the phase transition. Since we consider an expansion in the fluctuations, rather than the conventional expansion in the coupling constant, convergence to the exact results is found in both phases when taking the thermodynamic limit. PMID- 14525232 TI - Fluctuating dimension in a discrete model for quantum gravity based on the spectral principle. AB - The spectral principle of Connes and Chamseddine is used as a starting point to define a discrete model for Euclidean quantum gravity. Instead of summing over ordinary geometries, we consider the sum over generalized geometries where topology, metric, and dimension can fluctuate. The model describes the geometry of spaces with a countable number n of points, and is related to the Gaussian unitary ensemble of Hermitian matrices. We show that this simple model has two phases. The expectation value , the average number of points in the Universe, is finite in one phase and diverges in the other. We compute the critical point as well as the critical exponent of . Moreover, the space-time dimension delta is a dynamical observable in our model, and plays the role of an order parameter. The computation of is discussed and an upper bound is found, < 2. PMID- 14525233 TI - Large-N reduction in the continuum three-dimensional Yang-Mills theory. AB - Numerical and theoretical evidence leads us to propose the following: Three dimensional Euclidean Yang-Mills theory in the planar limit undergoes a phase transition on a torus of side l=l(c). For l>l(c) the planar limit is l independent, as expected of a noninteracting string theory. We expect the situation in four dimensions to be similar. PMID- 14525234 TI - Back-to-back correlations of high-p(T) hadrons in relativistic heavy-ion collisions. AB - We investigate the suppression factor and the azimuthal correlation function for high p(T) hadrons in central Au+Au collisions at sqrt[s(NN)]=200 GeV by using a dynamical model in which hydrodynamics is combined with explicitly traveling jets. We study the effects of parton energy loss in a hot medium, intrinsic k(T) of partons in a nucleus, and p (perpendicular) broadening of jets on the back-to back correlations of high p(T) hadrons. Parton energy loss is found to be a dominant effect on the reduction of the awayside peaks in the correlation function. PMID- 14525235 TI - High precision measurement of the superallowed 0+ --> 0+ beta decay of 22Mg. AB - The half-life, 3.8755(12) s, and superallowed branching ratio, 0.5315(12), for 22Mg beta decay have been measured with high precision. The latter depended on gamma-ray intensities being measured with an HPGe detector calibrated for relative efficiencies to an unprecedented 0.15%. Previous precise measurements of 0+ --> 0+ transitions have been restricted to the nine that populate stable daughter nuclei. No more such cases exist, and any improvement in a critical Cabibbo-Kobayashi-Maskawa unitarity test must depend on precise measurements of more exotic nuclei. With this branching-ratio measurement, we show those to be possible for T(z)=-1 parents. We obtain a corrected Ft value of 3071(9) s, in good agreement with expectations. PMID- 14525236 TI - Clarification of the three-body decay of 12C (12.71 MeV). AB - Using beta decays of a clean source of 12N produced at the IGISOL facility, we have measured the breakup of the 12C (12.71 MeV) state into three alpha particles with a segmented particle detector setup. The high quality of the data permits solving the question of the breakup mechanism of the 12.71 MeV state, a longstanding problem in few-body nuclear physics. Among existing models, a modified sequential model fits the data best, but systematic deviations indicate that a three-body description is needed. PMID- 14525237 TI - Demonstration of dispersion-canceled quantum-optical coherence tomography. AB - We present an experimental demonstration of quantum-optical coherence tomography. The technique makes use of an entangled twin-photon light source to carry out axial optical sectioning. It is compared to conventional optical coherence tomography. The immunity of the quantum version to dispersion, as well as a factor of 2 enhancement in resolution, is experimentally demonstrated. PMID- 14525238 TI - Cavity solitons in two-level lasers with dense amplifying medium. AB - Local-field effects are known to induce bistability in dense optical media. We examine theoretically whether this property is preserved in broad-area cavities, and show that bistability between the homogeneous lasing and nonlasing states of the system persists provided a Fourier filtering technique is used to prevent off axis emission. The resulting bistability gives rise to spatial light localization in the form of cavity solitons, which exhibit a particularly large degree of plasticity as a function of the characteristics of the addressing beam. This is the simplest laser able to sustain cavity solitons. PMID- 14525239 TI - Light pulse slowing down up to 0.025 cm/s by photorefractive two-wave coupling. AB - It is shown experimentally and theoretically that photorefractive wave coupling can be used for dramatic (< or approximately 0.025 cm/s) deceleration of light pulses whose width is larger than (or comparable with) the nonlinear response time. This classical nonlinear scheme exhibits similarities with the technique based on the quantum effect of electromagnetically induced transparency. The main distinctive feature of our scheme is amplification of the delayed output pulse. Advantages of the novel technique and its prospects for manipulation with light photons are discussed. PMID- 14525240 TI - Synchronization and oscillator death in oscillatory media with stirring. AB - The effect of stirring in an inhomogeneous oscillatory medium is investigated. We show that the stirring rate can control the macroscopic behavior of the system producing collective oscillations (synchronization) or complete quenching of the oscillations (oscillator death). We interpret the homogenization rate due to mixing as a measure of global coupling and compare the phase diagrams of stirred oscillatory media and of populations of globally coupled oscillators. PMID- 14525241 TI - Estimating good discrete partitions from observed data: symbolic false nearest neighbors. AB - A symbolic analysis of observed time series requires a discrete partition of a continuous state space containing the dynamics. A particular kind of partition, called "generating," preserves all deterministic dynamical information in the symbolic representation, but such partitions are not obvious beyond one dimension. Existing methods to find them require significant knowledge of the dynamical evolution operator. We introduce a statistic and algorithm to refine empirical partitions for symbolic state reconstruction. This method optimizes an essential property of a generating partition, avoiding topological degeneracies, by minimizing the number of "symbolic false nearest neighbors." It requires only the observed time series and is sensible even in the presence of noise when no truly generating partition is possible. PMID- 14525242 TI - Nusselt number measurements for turbulent Rayleigh-Benard convection. AB - We present high-precision measurements of the Nusselt number N as a function of the Rayleigh number R for a cylindrical sample of water (Prandtl number sigma=4.4) of height L approximately equal to 50 cm and aspect ratio Gamma identical with D/L approximately equal to 1 (D is the diameter) for 3 x 10(9)< or =R< or =6 x 10(10). For R approximately 3 x 10(9) the data are consistent with existing results for acetone (sigma=4.0, R< or =3 x 10(9)). There the measurements are also consistent with a model by Grossmann and Lohse (GL). As R increases, the measurements fall below the GL prediction. Near R=6 x 10(10) the prediction is 8% above the data. PMID- 14525243 TI - Interaction potential and hopping dynamics governing sliding friction. AB - The friction force on a nanometer-sized tip sliding on a surface is related to the thermally activated hopping of the contact atoms on an effective atomic interaction potential. A general analytical expression relates the height of this potential and the hopping attempt frequency to measurements of the velocity dependence of the friction force performed with an atomic force microscope. While the height of the potential is roughly proportional to the normal load, the attempt frequency falls in the range of mechanical eigenfrequencies of the probing tip in contact with the surface. PMID- 14525244 TI - Superstatistical mechanics of tracer-particle motions in turbulence. AB - The Lagrangian stochastic model of Reynolds [Phys. Fluids 15, L1-4 (2003)]] for the accelerations of fluid particles in turbulence is shown to predict precisely the observed Reynolds-number dependency of the distribution of Lagrangian accelerations and the exponents characterizing the observed extended self similarity scaling of the Lagrangian velocity structure functions. Departures from superstatistics of the log-normal kind are accounted for and their impact upon model predictions is quantified. PMID- 14525245 TI - Light intensification towards the Schwinger limit. AB - A method to generate ultrahigh intense electromagnetic fields is suggested, based on the laser pulse compression, carrier frequency upshift, and focusing by a counterpropagating breaking plasma wave, relativistic flying parabolic mirror. This method allows us to achieve the quantum electrodynamics critical field (Schwinger limit) with present-day laser systems. PMID- 14525246 TI - Space-charge-limited 2D electron flow between two flat electrodes in a strong magnetic field. AB - An approximate analytic solution is constructed for the 2D space-charge-limited emission by a cathode surrounded by nonemitting conducting ledges of width Lambda. An essentially exact solution (via conformal mapping) of the electrostatic problem in vacuum is matched to the solution of a linearized problem in the space charge region whose boundaries are sharp due to the presence of a strong magnetic field. The current density growth in a narrow interval near the edges of the cathode depends strongly on Lambda. We obtain an empirical formula for the total current as a function of Lambda which extends to more general cathode geometries. PMID- 14525247 TI - Characteristics of electron heat transport of plasma with an electron internal transport barrier in the large helical device. AB - Associated with the transition from ion root to electron root, an electron internal transport barrier (ITB) appears in the large helical device, when the heating power of electron cyclotron resonance heating exceeds the threshold power. The incremental thermal diffusivity of electron heat transport chi(inc)(e) in the ITB plasma is much lower than that in the plasma with the heating power below the threshold, and the thermal diffusivity chi(e) decreases with increasing of heating power [dchi(e)/d(P/n(e))<0] in helical ITB plasmas. PMID- 14525248 TI - Tokamak equilibria with reversed current density. AB - Observations of nearly zero toroidal current in the central region of tokamaks (the "current hole") raises the question of the existence of toroidal equilibria with very low or reversed current in the core. The solutions of the Grad Shafranov equilibrium equation with hollow toroidal current density profile including negative current density in the plasma center are investigated. Solutions of the corresponding eigenvalue problem provide simple examples of such equilibrium configurations. More realistic equilibria with toroidal current density reversal are computed using a new equilibrium problem formulation and computational algorithm which do not assume nested magnetic surfaces. PMID- 14525249 TI - Anomalous transmission of an ultrashort ionizing laser pulse through a thin foil. AB - The formation of a highly anisotropic photoelectron velocity distribution as a result of the interaction of a powerful ultrashort laser pulse with a thin foil is found to yield a large skin-layer depth and an anomalous increase of the transmission coefficient. The physical reason for the effect is the influence of the incident wave magnetic field, through the Lorenz force, on the electron kinetics in the skin layer. PMID- 14525250 TI - Quantum transmission of atoms through a slab of superfluid helium. AB - We describe a measurement of the transmission probability of 4He atoms through a freely suspended slab of superfluid 4He at low temperatures. In our experiment the slab is realized by using an array of parallel cylindrical holes of diameter 51 microm in a glass disc of thickness 190 microm. By controlling the chemical potential, the holes can be made to fill or empty with liquid, and the surface curvature varied. We have measured the transmission of atom beams, generated by a thin-film heater and detected with a sensitive bolometer, through this structure. The results show that the dominant transmission channel is atom-R+ roton-atom with a probability p approximately 0.12 and that R+ rotons can undergo total internal reflection at the free liquid surfaces. PMID- 14525251 TI - Effective mass of two-dimensional 3He. AB - We use structural information from diffusion Monte Carlo calculations for two dimensional 3He to calculate the effective mass. Static effective interactions are constructed from the density and spin-structure functions using sum rules. We find that both spin and density fluctuations contribute about equally to the effective mass. Our results show, in agreement with recent experiments, a flattening of the single-particle self-energy with increasing density, which eventually leads to a divergent effective mass. PMID- 14525252 TI - Specific Raman signatures of a dimetallofullerene peapod. AB - We report on the Raman spectroscopy of a dimetallofullerene peapod, (La(2)@C(80))(m)@SWNT. Drastic changes are observed with respect to pristine nanotubes. A sharp intense line at 142 cm(-1) is interpreted as a signature of polymerization of the encapsulated metallofullerenes. Additional strong signatures appear at about 400, 520, and 640 cm(-1), respectively. Their intensity suggests the existence of an enhancement effect. The stiffening and the up-shift of the G-band modes appear to imply that a charge transfer process between the nanotube and the peas occurs. PMID- 14525253 TI - Multilayer adsorption and wetting of acetone on graphite. AB - Adsorption/desorption isotherms of acetone on highly oriented pyrolytic graphite have been measured by ellipsometry for temperatures above the bulk triple point. The behavior in the monolayer and submonolayer regime is conventional, with 2D gas-liquid and 2D liquid-solid coexistence regions. Further liquid monolayers grow on top of the completed monolayer. The growth is basically layer-by-layer. For temperatures between 190 K and the triple point a prewetting-type transition occurs with a thin-thick jump of the layer thickness on adsorption but a layer wise removal of the film on desorption. In this temperature regime the first monolayer is solid and its molecules are oriented perpendicular to the substrate whereas the higher layers are orientationally disordered polar liquid. PMID- 14525255 TI - Generation of coherent zone boundary phonons by impulsive excitation of molecules. AB - A coherent zone boundary phonon (ZBP) of solid Kr (f(m)=1.54 THz) is observed in ultrafast pump-probe spectra of I2 guest molecules in a Kr crystal. Its phase is stable for at least 10 ps. The femtosecond pump pulse induces an electronic transition in I2. The resulting expansion of the guest's electronic cloud impulsively excites phonons in the host. Detection at the impurity after some picoseconds selects the ZBP due to its low group velocity. A ZBP amplitude of 0.02 A is estimated. PMID- 14525254 TI - Stereo-selective swelling of imprinted cholesteric networks. AB - Molecular chirality, and the chiral symmetry breaking of resulting macroscopic phases, can be topologically imprinted and manipulated by cross-linking and swelling of polymer networks. We present a new experimental approach to stereo specific separation of chiral isomers by using a cholesteric elastomer in which a helical director distribution has been topologically imprinted by cross-linking. This makes the material unusual in that is has a strong phase chirality, but no molecular chirality at all; we study the nature and parameters controlling the twist-untwist transition. Adding a racemic mixture to the imprinted network results in selective swelling by only the component of "correct" handedness. We investigate the capacity of demixing in a racemic environment, which depends on network parameters and the underlying nematic order. PMID- 14525256 TI - Mixing effects for the structural relaxation in binary hard-sphere liquids. AB - We report extensive molecular-dynamics-simulation results for binary mixtures of hard spheres for different size disparities and different mixing percentages, for packing fractions up to 0.605, and over a characteristic time interval spanning up to 5 orders in magnitude. We explore the changes in the evolution of glassy dynamics due to mixing and discover two opposite scenarios: For large size disparity, increasing the mixing percentage of small particles leads to a speed up of long-time dynamics, while small disparity leads to a slowing down. These results agree with predictions based on the mode-coupling theory for ideal-glass transitions. PMID- 14525257 TI - Driving rate effects on crackling noise. AB - Many systems respond to slowly changing external conditions with crackling noise, created by avalanches or pulses with a broad range of sizes. Examples range from Barkhausen noise (BN) in magnets to earthquakes. In this Letter, we discuss the effects of increasing driving rate Omega on the scaling behavior of the avalanche size and duration distributions as well as qualitative effects of Omega on the power spectra. We derive an exponent inequality as a criteria for the relevance of Omega. To illustrate these general results, we use recent experiments on BN as a successful example. PMID- 14525258 TI - Glass transition in ultrathin polymer films: calorimetric study. AB - The ultrasensitive differential scanning calorimetry is used to observe the glass transition in thin (1-400 nm) spin-cast films of polystyrene, poly (2-vinyl pyridine) and poly (methyl methacrylate) on a platinum surface. A pronounced glass transition is observed even at a thickness as small as 1-3 nm. Using the high heating (20-200 K/ms) and cooling (1-2 K/ms in glass transition region) rates which are typical for this technique, we do not observe appreciable dependence of the glass transition temperature over the thickness range from hundreds of nanometers down to 3 nm thick films. The evolution of calorimetric data with film thickness is discussed in terms of broadening of transition dynamics and loss of transition contrast. PMID- 14525259 TI - Fluctuation mediated interaction and phase separation of nanoparticles in a liquid crystal solvent. AB - Water-in-oil microemulsions of nanodroplets in the isotropic phase of a thermotropic liquid crystal exhibit, with decreasing temperature and in anticipation of a demixing transition, enhanced correlation in fluctuations of both molecular orientation and droplet concentration. Mean field modeling of this pretransition behavior, on the basis of a lattice in which the nanodroplets are introduced as holes, shows that the observed interdroplet attractive interaction is produced by the disordering effect of the droplets on the liquid crystal and mediated solely by paranematic fluctuations. PMID- 14525260 TI - Interface profiles near three-phase contact lines in electric fields. AB - Long-range electrostatic fields deform the surface profile of a conductive liquid in the vicinity of the contact line. We have investigated the equilibrium profiles by balancing electrostatic and capillary forces locally at the liquid vapor interface. Numerical results show that the contact angle at the contact line approaches Young's angle. Simultaneously, the local curvature displays a weak algebraic divergence. Furthermore, we present an asymptotic analytical model, which confirms these results and elucidates the scaling behavior of the profile close to the contact line. PMID- 14525261 TI - Novel surface tension isotherm for surfactants based on local density functional theory. AB - In this Letter we report a new general method for calculating of surface tension isotherms in the presence of surfactants, based on a local density functional. We illustrate this method by deriving the interfacial tension isotherm for nonionic surfactants at an air-water or oil-water interface by using the self-consistent field theory of polymer brushes. We consider a particular case of local density functional to calculate explicitly how the interfacial tension and the surfactant adsorption depend on the surfactant bulk concentration. Experimental data for the surface tension and the surfactant adsorption isotherm for nonionic surfactants were interpreted with the help of the new isotherm. Very good agreement between the adsorption of n-dodecyl pentaoxyethylene glycol ether (C12E5) at an air-water interface, calculated from the surface tension isotherm and small-angle neutron scattering is obtained. PMID- 14525262 TI - Infinite family of persistence exponents for interface fluctuations. AB - We show experimentally and theoretically that the persistence of large deviations in equilibrium step fluctuations is characterized by an infinite family of independent exponents. These exponents are obtained by carefully analyzing dynamical experimental images of Al/Si(111) and Ag(111) equilibrium steps fluctuating at high (970 K) and low (320 K) temperatures, respectively, and by quantitatively interpreting our observations on the basis of the corresponding coarse-grained discrete and continuum theoretical models for thermal surface step fluctuations under attachment/detachment ("high-temperature") and edge-diffusion limited kinetics ("low-temperature"), respectively. PMID- 14525263 TI - Imaging the internal electronic structure of a surface adsorbate with low energy ions. AB - Time-of-flight spectra were collected for 2.5 keV 7Li+ backscattered from Fe surfaces covered with submonolayers of iodine. Li singly scattered from the adatoms has a consistently larger neutral fraction than for scattering from the substrate, implying a region of positive charge atop the iodine. The neutral fraction decreases for off-normal exit angles, indicating a nonuniform charge distribution around the polarizable adsorbates. This demonstrates that ion scattering can image the internal electronic structure of an adatom and provides an explanation for anomalous work function changes. PMID- 14525264 TI - Self-similar structures near boundaries in strained systems. AB - We study the buckling of thin elastic plates caused by residual strains concentrated near a free edge. This is a model for plant leaves and torn plastic sheet morphologies. We derive new governing equations explaining self-similar patterns reported earlier in experiments. We reveal the cascade mechanism, determine the bounds for its wavelengths, and predict a similarity factor of 3 in agreement with experiments. This is confirmed by numerical solutions with up to five generations of wrinkles. PMID- 14525265 TI - Radiation-induced "zero-resistance state" and the photon-assisted transport. AB - We demonstrate that the radiation-induced "zero-resistance state" observed in a two-dimensional electron gas is a result of the nontrivial structure of the density of states of the systems and the photon-assisted transport. A toy model of a quantum tunneling junction with oscillatory density of states in leads catches most of the important features of the experiments. We present a generalized Kubo-Greenwood conductivity formula for the photon-assisted transport in a general system and show essentially the same nature of the transport anomaly in a uniform system. PMID- 14525266 TI - Spin excitations of the spin-polarized electron gas in semimagnetic quantum wells. AB - Collective and single-particle spin-flip excitations of a two-dimensional electron gas in a semimagnetic Cd(1-x)Mn(x)Te quantum well are observed by resonant Raman scattering. Application of a magnetic field splits the spin subbands and a spin polarization is induced in the electron gas. Above 1 T the collective modes, which disperse with the in-plane wave vector, dominate the spectra. The local spin-density approximation provides a good description of our results and enables us to confirm that the energy of the low wave vector collective mode is given by the bare Zeeman energy. PMID- 14525267 TI - Radiation-induced magnetoresistance oscillations in a 2D electron gas. AB - Recent measurements of a 2D electron gas subjected to microwave radiation reveal a magnetoresistance with an oscillatory dependence on the ratio of radiation frequency to cyclotron frequency. We perform a diagrammatic calculation and find radiation-induced resistivity oscillations with the correct period and phase. Results are explained via a simple picture of current induced by photoexcited disorder-scattered electrons. The oscillations increase with radiation intensity, easily exceeding the dark resistivity and resulting in negative-resistivity minima. At high intensity, we identify additional features, likely due to multiphoton processes, which have yet to be observed experimentally. PMID- 14525268 TI - Magnetic penetration depth measurements of Pr2-xCexCuO4-delta films on buffered substrates: evidence for a nodeless gap. AB - We report measurements of the inverse squared magnetic penetration depth, lambda( 2)(T), in Pr(2-x)Ce(x)CuO(4-delta) (0.115< or =x < or =0.152) superconducting films grown on SrTiO3 (001) substrates coated with a buffer layer of insulating Pr2CuO4. lambda(-2)(0), T(c), and normal-state resistivities of these films indicate that they are clean and homogeneous. Over a wide range of Ce doping, 0.124< or =x < or =0.144, lambda(-2)(T) at low T is flat: it changes by less than 0.15% over a factor of 3 change in T, indicating a gap in the superconducting density of states. Fits to the first 5% decrease in lambda(-2)(T) produce values of the minimum superconducting gap in the range of 0.29< or =Delta(min)/k(B)T(c)< or =1.01. PMID- 14525269 TI - Density functional study of the phase diagram and pressure-induced superconductivity in p: implication for spintronics. AB - The high-pressure phase diagram of P is studied using density functional total energy, linear response lattice dynamics and model Debye-Gruneisen theories. The volume dependent electron-phonon coupling lambda approximately 0.7-0.9 is extracted for the bcc structure and found to increase with increasing volume. We propose that this phase might be realized in epitaxial thin films using templates such as V(100), Fe(100), or Cr(100) relevant to spintronics applications. PMID- 14525270 TI - Scalings of domain wall energies in two dimensional Ising spin glasses. AB - We study domain wall energies of two dimensional spin glasses. The scaling of these energies depends on the model's distribution of quenched random couplings, falling into three different classes. The first class is associated with the exponent theta approximately -0.28; the other two classes have theta=0, as can be justified theoretically. In contrast to previous claims, we find that theta=0 does not indicate d=d(c)(l) but rather d< or =d(c)(l), where d(c)(l) is the lower critical dimension. PMID- 14525271 TI - Extended versus local fluctuations in quantum critical Ce(Ru1-xFex)2Ge2 (x=xc=0.76). AB - We present inelastic neutron scattering experiments, performed near the antiferromagnetic quantum critical point in Ce(Ru0.24Fe0.76)2Ge2. Both local and long-range fluctuations of the local moments are observed, but due to the Kondo effect only the latter are critical. We propose a phenomenological expression which fits the energy E, temperature T, and wave vector q dependences of the dynamic susceptibility, describing the non-Fermi liquid E/T scaling found at every q. PMID- 14525272 TI - Ferromagnetism of MnAs studied by heteroepitaxial films on GaAs(001). AB - Thin epitaxial films of MnAs--promising candidates for the spin injection into semiconductors--are well known to undergo simultaneously a first-order structural and magnetic phase transition at 10-40 degrees C. The evolution of stress and magnetization of MnAs/GaAs(001), both measured quantitatively with our cantilever beam magnetometer at the coexistence region of alpha-MnAs and beta-MnAs, reveal an orthorhombically distorted unit cell of the ferromagnetic phase, which provides important clues on the origin of ferromagnetism in MnAs. PMID- 14525273 TI - String order in half-integer-spin antiferromagnetic Heisenberg chains. AB - We derive the extended string order parameter O(S) for antiferromagnetic Heisenberg chains with half-integer spin S in the valence-bond-solid picture. We obtain the analytic power-law scaling of O(S) versus the chain length L and show that O(S) scales at an extremely slow pace that decreases rapidly with growing spin magnitude. Furthermore, accurate numerical calculations show that the power law scaling sets in only when L exceeds a characteristic length scale l(S) which increases very fast with growing S. Consequently, a pseudo-long-range string order exists in half-integer-spin Heisenberg chains. The implications of this result and its relationship to other topological features such as the end-chain states are discussed. PMID- 14525274 TI - Probing hole-induced ferromagnetic exchange in magnetic semiconductors by inelastic neutron scattering. AB - The effect of hole doping on the exchange coupling of the nearest neighbor (NN) Mn pairs in Zn(1-x)MnxTe is probed by inelastic neutron scattering. The difference in the NN exchange energy DeltaJ1 in the presence and in the absence of the holes is determined. The obtained value of DeltaJ1 is in good agreement with the predictions of the Zener/RKKY model, even on the insulator side of the metal-insulator transition. PMID- 14525275 TI - Unusual symmetries in the Kugel-Khomskii Hamiltonian. AB - The Kugel-Khomskii Hamiltonian for cubic titanates describes spin and orbital superexchange interactions between d(1) ions having threefold degenerate t(2g) orbitals. Since orbitals do not couple along "inactive" axes, perpendicular to the orbital planes, the total number of electrons in |alpha> orbitals in any such plane and the corresponding total spin are both conserved. A Mermin-Wagner construction shows that there is no long-range spin ordering at nonzero temperatures. Inclusion of spin-orbit coupling allows such ordering, but even then the excitation spectrum is gapless due to a continuous symmetry. Thus, the observed order and gap require more symmetry breaking terms. PMID- 14525276 TI - Collective density-wave excitations in two-leg Sr14-xCaxCu24O41 ladders. AB - Raman measurements in the 1.5-20 cm(-1) energy range were performed on single crystals of Sr14-xCaxCu24O41. A quasielastic scattering peak (QEP) which softens with cooling is observed only in the polarization parallel to the ladder direction for samples with x=0, 8, and 12. The QEP is a Raman fingerprint of pinned collective density wave excitations screened by uncondensed carriers in the ladder structures. Our results suggest that transport in metallic samples, which is similar to transport in underdoped high-T(c) cuprates, is driven by a collective electronic response. PMID- 14525277 TI - High-energy phonon branches of an individual metallic carbon nanotube. AB - We present excitation-energy dependent Raman measurements between 2.05 and 2.41 eV on the same individual carbon nanotube. We find a change in the Raman frequencies of both the D mode (63 cm(-1)/eV) and the high-energy modes. The observed frequencies of the modes at approximately 1600 cm(-1) as a function of laser-energy map the phonon dispersion relation of a metallic tube near the Gamma point of the Brillouin zone. Our results prove the entire first-order Raman spectrum in single-wall carbon nanotubes to originate from double-resonant scattering. Moreover, we confirm experimentally the phonon softening in metallic tubes by a Peierls-like mechanism. PMID- 14525278 TI - Decoherence-free subspaces in quantum key distribution. AB - We demonstrate that two recent innovations in the field of practical quantum key distribution (one-way autocompensation and passive detection) are closely related to the methods developed to protect quantum computations from decoherence. We present a new scheme that combines these advantages, and propose a practical implementation of this scheme that is feasible using existing technology. PMID- 14525279 TI - Operationally invariant measure of the distance between quantum states by complementary measurements. AB - We propose an operational measure of distance of two quantum states, which conversely tells us their closeness. This is defined as a sum of differences in partial knowledge over a complete set of mutually complementary measurements for the two states. It is shown that the measure is operationally invariant and it is equivalent to the Hilbert-Schmidt distance. The operational measure of distance provides a remarkable interpretation of the information distance between quantum states. PMID- 14525280 TI - Lower bound for electron spin entanglement from beam splitter current correlations. AB - We determine a lower bound for the entanglement of formation of pairs of electron spins injected into a mesoscopic conductor. The bound can be expressed in terms of experimentally accessible quantities, the zero-frequency current correlators (shot noise power or cross correlators) after transmission through an electronic beam splitter and can be used to gain information about the entanglement from experiment. Spin relaxation (T1 processes) and decoherence (T2) during the ballistic coherent transmission of carriers are taken into account within Bloch theory. A variable inhomogeneous magnetic field gives rise to a useful lower bound for the entanglement of arbitrary states. The decrease in entanglement due to thermally mixed states is studied. Both the entanglement of the output of a source (entangler) and T(1,2) can be determined from current correlators. PMID- 14525281 TI - Electrical manipulation of nanomagnets. AB - We demonstrate that it is possible to manipulate the magnetic coupling between two nanomagnets by means of an ac electric field. In the scheme suggested, the magnetic coupling is mediated by a magnetic particle that is in contact with both nanomagnets via tunnel barriers. The time-dependent electric field is applied so that the height of first one barrier then the other is suppressed in an alternating fashion. We show that the result is a pumping of magnetization from one nanomagnet to the other through the mediating particle. The dynamics of the magnetization of the mediating particle allows the coupling to be switched between being ferromagnetic and being antiferromagnetic. PMID- 14525282 TI - Comment on "Quasicrystal-crystal transformation in Zn-Mg-rare-earth alloys". PMID- 14525284 TI - Comment on "Ground-state phase diagram of a half-filled one-dimensional extended Hubbard model". PMID- 14525286 TI - Comment on "Conservative quantum computing". PMID- 14525288 TI - Towards quantum superpositions of a mirror. AB - We propose an experiment for creating quantum superposition states involving of the order of 10(14) atoms via the interaction of a single photon with a tiny mirror. This mirror, mounted on a high-quality mechanical oscillator, is part of a high-finesse optical cavity which forms one arm of a Michelson interferometer. By observing the interference of the photon only, one can study the creation and decoherence of superpositions involving the mirror. A detailed analysis of the requirements shows that the experiment is within reach using a combination of state-of-the-art technologies. PMID- 14525289 TI - Ray helicity: geometric invariant for multidimensional resonant wave conversion. AB - For a multicomponent wave field propagating into a multidimensional conversion region, the rays are shown to be helical, in general. For a ray-based quantity to have a fundamental physical meaning, it must be invariant under the following two groups of transformations, which are used to construct solutions: congruence transformations (which involve linear combinations of components of the multicomponent wave field) and canonical transformations (which act on the ray phase space). It is shown that for conversion between two waves there is a new invariant not previously discussed: the intrinsic helicity of the ray. PMID- 14525290 TI - Local quantum criticality in confined fermions on optical lattices. AB - Using quantum Monte Carlo simulations, we show that the one-dimensional fermionic Hubbard model in a harmonic potential displays quantum critical behavior at the boundaries of a Mott-insulating region. A local compressibility defined to characterize the Mott-insulating phase has a nontrivial critical exponent. Both the local compressibility and the variance of the local density show universality with respect to the confining potential. We determine a generic phase diagram, which allows the prediction of the phases to be observed in experiments with ultracold fermionic atoms trapped on optical lattices. PMID- 14525291 TI - Supersolid versus phase separation in atomic bose-fermi mixtures. AB - We show that a two-dimensional atomic mixture of bosons and fermions cooled into their quantum degenerate states and subject to an optical lattice develops a supersolid phase characterized by the simultaneous presence of a nontrivial crystalline order and phase order. This transition is in competition with phase separation. We determine the phase diagram of the system and propose an experiment allowing for the observation of the supersolid phase. PMID- 14525292 TI - Calculation of the scattering amplitude from discrete eigenvalues. AB - We show that the scattering amplitude for a target with spherical symmetry can be directly calculated from the discrete eigenvalues of the corresponding stationary wave equation solved under an absorbing boundary condition, and thus uncover a direct connection between the scattering amplitude and the wave number spectrum. As an illustration, we apply this approach to the scattering of an electromagnetic wave by an array of dielectric slabs and find the transmission coefficient of the array to be completely determined by the spacing between adjacent eigenvalues. PMID- 14525293 TI - Negative specific heat, the thermodynamic limit, and ergodicity. AB - Thermodynamic stability, in particular, the positivity of the specific heat in the microcanonical ensemble, is not an automatic consequence of the thermodynamic limit. But it holds under special circumstances such as for the most important case of quantum-mechanical Coulomb systems. Therefore, it is surprising that there are experimental indications to the contrary. In this Letter we study a simple model for which the microcanonical specific heat is positive, if the system is ergodic. However, if the system is not ergodic, the energy shell in phase space has some ergodic components with a negative specific heat. This provides another possible general pathway for a negative specific heat in addition to the commonly accepted, the small number of particles. PMID- 14525294 TI - Quantum to classical transition for random walks. AB - We look at two possible routes to classical behavior for the discrete quantum random walk on the integers: decoherence in the quantum "coin" which drives the walk, or the use of higher-dimensional (or multiple) coins to dilute the effects of interference. We use the position variance as an indicator of classical behavior and find analytical expressions for this in the long-time limit; we see that the multicoin walk retains the "quantum" quadratic growth of the variance except in the limit of a new coin for every step, while the walk with decoherence exhibits "classical" linear growth of the variance even for weak decoherence. PMID- 14525295 TI - Levy model for interstellar scintillations. AB - Observations of radio signals from distant pulsars provide a valuable tool for investigation of interstellar turbulence. The time shapes of the signals are the result of pulse broadening by the fluctuating electron density in the interstellar medium. While the scaling of the shapes with the signal frequency is well understood, the observed anomalous scaling with respect to the pulsar distance has remained a puzzle for more than 30 years. We propose a new model for interstellar electron density fluctuations, which explains the observed scaling relations. We suggest that these fluctuations obey Levy statistics rather than Gaussian statistics, as assumed in previous treatments of interstellar scintillations. PMID- 14525296 TI - Horizon ratio bound for inflationary fluctuations. AB - We demonstrate that the gravity wave background amplitude implies a robust upper bound on the wavelength-to-horizon ratio at the end of inflation: lambda/H(-1) less than or approximately equal e(60), as long as the cosmic energy density does not drop faster than radiation subsequent to inflation. This limit implies that N, the number of e-folds between horizon exit and the end of inflation for wave modes of interest, is less, similar 60 plus a model-dependent factor-for vast classes of slow-roll models, N less than or approximately equal 67. As an example, this bound solidifies the tension between observations of the cosmic microwave background anisotropies and chaotic inflation with a phi(4) potential by closing the escape hatch of large N (<62). PMID- 14525297 TI - Correlated adiabatic and isocurvature cosmic microwave background fluctuations in the wake of the results from the wilkinson microwave anisotropy probe. AB - In general correlated models, in addition to the usual adiabatic component with a spectral index n(ad1) there is another adiabatic component with a spectral index n(ad2) generated by entropy perturbation during inflation. We extend the analysis of a correlated mixture of adiabatic and isocurvature cosmic microwave background fluctuations of the Wilkinson Microwave Anisotropy Probe (WMAP) group, who set the two adiabatic spectral indices equal. Allowing n(ad1) and n(ad2) to vary independently we find that the WMAP data favor models where the two adiabatic components have opposite spectral tilts. Using the WMAP data only, the 2sigma upper bound for the isocurvature fraction f(iso) of the initial power spectrum at k(0)=0.05 Mpc(-1) increases somewhat, e.g., from 0.76 of n(ad2)=n(ad1) models to 0.84 with a prior n(iso)<1.84 for the isocurvature spectral index. PMID- 14525299 TI - A composite chiral pair of rotational bands in the odd-A nucleus 135Nd. AB - High-spin states in 135Nd were populated with the 110Pd(30Si,5n)135Nd reaction at a 30Si bombarding energy of 133 MeV. Two DeltaI=1 bands with close excitation energies and the same parity were observed. These bands are directly linked by DeltaI=1 and DeltaI=2 transitions. The chiral nature of these two bands is confirmed by comparison with three-dimensional tilted axis cranking calculations. This is the first observation of a three-quasiparticle chiral structure and establishes the primarily geometric nature of this phenomenon. PMID- 14525300 TI - Phase transitions in angle variables. AB - Phase transitions in angle variables are studied. An example of angular phase transition, an axially to triaxially deformed "shape" transition in nuclei, is discussed. Spectroscopic signatures for the occurrence of these transitions are suggested. Preliminary experimental evidence is presented. PMID- 14525301 TI - Derivation of the density functional theory from the cluster expansion. AB - The density functional theory is derived from a cluster expansion by truncating the higher-order correlations in one and only one term in the kinetic energy. The formulation allows self-consistent calculation of the exchange correlation effect without imposing additional assumptions to generalize the local density approximation. The pair correlation is described as a two-body collision of bound state electrons, and modifies the electron- electron interaction energy as well as the kinetic energy. The theory admits excited states, and has no self interaction energy. PMID- 14525302 TI - Probing electronic states of Ne2+ and Ar2+ by measuring kinetic-energy-release distributions. AB - Dissociative decay of metastable, electronically excited neon and argon dimer ions produces fragment ions with strikingly dissimilar kinetic-energy-release distributions. The distributions have been modeled based on ab initio calculations of potential energy curves. The unusual bimodal distribution observed for dissociation of Ne2+ arises from competition between radiative and nonradiative decay of the long-lived II(1/2)(u) state. For Ar2+, however, electronic predissociation is insignificant. PMID- 14525303 TI - Reversed Doppler effect in photonic crystals. AB - Nonrelativistic reversed Doppler shifts have never been observed in nature and have only been speculated to occur in pathological systems with simultaneously negative effective permittivity and permeability. This Letter presents a different, new physical phenomenon that leads to a nonrelativistic reversed Doppler shift in light. It arises when light is reflected from a moving shock wave propagating through a photonic crystal. In addition to reflection of a single frequency, multiple discrete reflected frequencies or a 10 GHz periodic modulation can also be observed when a single carrier frequency of wavelength 1 microm is incident. PMID- 14525304 TI - Observation of locked optical kink-antikink spatial shock waves. AB - We report the first experimental observation of optical spatial shock-wave pairs. The shock waves consist of two coupled kink and antikink beams that remain locked to each other throughout propagation in a nonlinear diffusion-driven photorefractive crystal. These coupled shock-wave pairs move undistorted at angles that fall outside their original angular sector of propagation. PMID- 14525305 TI - Negative lateral shift of a light beam transmitted through a dielectric slab and interaction of boundary effects. AB - It is found that when a light beam travels through a slab of optically denser dielectric medium in air, the lateral shift of the transmitted beam can be negative. This is a novel phenomenon that is reversed in comparison with the geometrical optic prediction according to Snell's law of refraction. A Gaussian shaped beam is analyzed in the paraxial approximation, and a comparison with numerical simulations is made. Finally, an explanation for the negativity of the lateral shift is suggested, in terms of the interaction of boundary effects of the slab's two interfaces with air. PMID- 14525306 TI - Shot noise in chaotic cavities from action correlations. AB - We consider universal shot noise in ballistic chaotic cavities from a semiclassical point of view and show that it is due to action correlations within certain groups of classical trajectories. Using quantum graphs as a model system, we sum these trajectories analytically and find agreement with random-matrix theory. Unlike all action correlations which have been considered before, the correlations relevant for shot noise involve four trajectories and do not depend on the presence of any symmetry. PMID- 14525307 TI - Why are chaotic attractors rare in multistable systems? AB - We show that chaotic attractors are rarely found in multistable dissipative systems close to the conservative limit. As we approach this limit, the parameter intervals for the existence of chaotic attractors as well as the volume of their basins of attraction in a bounded region of the state space shrink very rapidly. An important role in the disappearance of these attractors is played by particular points in parameter space, namely, the double crises accompanied by a basin boundary metamorphosis. Scaling relations between successive double crises are presented. Furthermore, along this path of double crises, we obtain scaling laws for the disappearance of chaotic attractors and their basins of attraction. PMID- 14525308 TI - Intermediate wave function statistics. AB - We calculate statistical properties of the eigenfunctions of two quantum systems that exhibit intermediate spectral statistics: star graphs and Seba billiards. First, we show that these eigenfunctions are not quantum ergodic, and calculate the corresponding limit distribution. Second, we find that they can be strongly scarred, in the case of star graphs by short (unstable) periodic orbits and, in the case of Seba billiards, by certain families of orbits. We construct sequences of states which have such a limit. Our results are illustrated by numerical computations. PMID- 14525309 TI - Strange attractors are classified by bounding Tori. AB - There is at present a doubly discrete classification for strange attractors of low dimension, d(L)<3. A branched manifold describes the stretching and squeezing processes that generate the strange attractor, and a basis set of orbits describes the complete set of unstable periodic orbits in the attractor. To this we add a third discrete classification level. Strange attractors are organized by the boundary of an open set surrounding their branched manifold. The boundary is a torus with g holes that is dressed by a surface flow with 2(g-1) singular points. All known strange attractors in R3 are classified by genus, g, and flow type. PMID- 14525310 TI - Kink-induced transport and segregation in oscillated granular layers. AB - We use experiments and molecular dynamics simulations of vertically oscillated granular layers to study horizontal particle segregation induced by a kink (a boundary between domains oscillating out of phase). Counterrotating convection rolls carry the larger particles in a bidisperse layer along the granular surface to a kink, where they become trapped. The convection originates from avalanches that occur inside the layer, along the interface between solidified and fluidized grains. The position of a kink can be controlled by modulation of the container frequency, making possible systematic harvesting of the larger particles. PMID- 14525311 TI - New tokamak plasma regime with stationary temperature oscillations. AB - During noninductively driven discharges in the Tore Supra tokamak, steady sinusoidal oscillations of the central electron temperature, lasting as long as 2 min, have been observed for the first time. Having no helical structure, they cannot be ascribed to any known MHD instability. The most plausible explanation of this new phenomenon is that the plasma current density and the electron temperature evolve as a nonlinearly coupled predator-prey system. This interpretation is supported by the numerical solution of coupled resistive current diffusion and heat transport equations. PMID- 14525312 TI - Kelvin-wave cascade on a vortex in superfluid 4He at a very low temperature. AB - A study by computer simulation is reported of the behavior of a quantized vortex line at a very low temperature when there is continuous excitation of low frequency Kelvin waves. There is no dissipation except by phonon radiation at a very high frequency. It is shown that nonlinear coupling leads to a net flow of energy to higher wave numbers and to the development of a simple spectrum of Kelvin waves that is insensitive to the strength and frequency of the exciting drive. The results are likely to be relevant to the decay of turbulence in superfluid 4He at very low temperatures. PMID- 14525313 TI - Phonon instabilities and the ideal strength of aluminum. AB - We have calculated the phonon spectra of aluminum as a function of strain using density functional perturbation theory for <110>, <100>, and <111> uniaxial tension, as well as relaxed <112>[111] shear. In all four cases, phonon instabilities occur at points away from the center of the Brillouin zone and intrude before the material becomes unstable according to elastic stability criteria. This is the first time the ideal strength of a metal has been shown to be dictated by instabilities in the acoustic phonon spectra. We go on to describe the crystallography of the unstable modes, all of which are shear in character. This work further suggests that shear failure is an inherent property of aluminum even in an initially dislocation-free perfect crystal. PMID- 14525314 TI - Nanocluster formation in electron-irradiated Li2O crystals observed by elastic diffuse neutron scattering. AB - Previous work performed on electron-irradiated Li2O crystals has demonstrated the simultaneous formation of two populations of colloids of metallic lithium, one is associated with oxygen bubbles and a typical size of >10 microm, while the other one consists of nanoclusters in the range of <10 nm. In the present neutron scattering investigation these small colloids are characterized in detail based on a thorough analysis of distortion scattering around Bragg peaks. It is shown that the small lithium colloids are slightly elongated precipitates with typical dimensions of around approximately 5 nm. For the large lithium colloids a well defined orientation relation with respect to the Li2O matrix has been determined. PMID- 14525315 TI - Predicting crystal structures with data mining of quantum calculations. AB - Predicting and characterizing the crystal structure of materials is a key problem in materials research and development. It is typically addressed with highly accurate quantum mechanical computations on a small set of candidate structures, or with empirical rules that have been extracted from a large amount of experimental information, but have limited predictive power. In this Letter, we transfer the concept of heuristic rule extraction to a large library of ab initio calculated information, and we demonstrate that this can be developed into a tool for crystal structure prediction. PMID- 14525316 TI - Microscopic description of the irradiation-induced amorphization in silicon. AB - We have investigated the atomistic mechanism behind the irradiation-induced amorphization in Si using molecular dynamics simulation techniques. The microscopic description of the process is based on the defect known as bond defect or IV pair. IV pairs recombine very fast when isolated, but if they interact to each other they survive longer times and thus accumulate giving rise to amorphization. This fact accounts for the superlinear behavior of the accumulated damage with dose and the different activation energies for recrystallization observed in the experiments. The molecular dynamics results have been used to define an atomistic model for amorphization and recrystallization which has been implemented in a kinetic Monte Carlo code. The model is able to reproduce quantitatively the dependence of the critical crystal amorphous transition on the irradiation parameters. PMID- 14525317 TI - Is there icosahedral ordering in liquid and undercooled metals? AB - The local structure of simple liquids is significantly different from that of corresponding crystalline systems. Signatures of fivefold local ordering have been previously found, but current knowledge is limited to pair distribution, leaving considerable uncertainty in the determination of the geometrical structure. New x-ray absorption experimental results on liquid and undercooled liquid copper, interpreted using an advanced data-analysis method based on multiple-scattering simulations, are shown to contain direct information on triplet correlations making feasible a reliable determination of the bond-angle distribution and fraction of nearly icosahedral configurations in liquids. PMID- 14525318 TI - Dynamics of a disordered, driven zero-range process in one dimension. AB - We study a disordered, driven zero range process which models a closed system of attractive particles that hop with site-dependent rates and whose steady state shows a condensation transition with increasing density. We characterize the dynamical properties of the mass fluctuations in the steady state in one dimension both analytically and numerically and show that there is a dynamic phase transition in the density-disorder plane. We also determine the form of the scaling function which describes the growth of the condensate as a function of time, starting from a uniform density distribution. PMID- 14525319 TI - Ab initio simulations of homoepitaxial SiC growth. AB - We present first-principle calculations on the initial stages of SiC homoepitaxial growth on the beta-SiC(111)-(sqrt[3]xsqrt[3]) surface. We show that the nonstoichiometric reconstruction plays a relevant role in favoring the attainment of high-quality films. The motivation is twofold: On one hand, we find that the reconstruction controls the kinetics of adatom incorporation; on the other hand, we observe that the energy gain upon surface stability can induce the reorganization of the deposited material into a crystalline structure, thus revealing that a surface-driven mechanism is able to stabilize defect-free layer deposition on Si-rich surfaces. PMID- 14525320 TI - Dynamic scaling, island size distribution, and morphology in the aggregation regime of submonolayer pentacene films. AB - Scaling behavior of the island size distribution through a universal scaling function f(u) is demonstrated for submonolayer pentacene islands in the aggregation regime (0.1D(*+)D(*-). AB - We present a measurement of time-dependent CP asymmetries and an updated determination of the CP-odd fraction in the decay B0-->D(*+)D(*-) using a data sample of 88x10(6)BB pairs collected by the BABAR detector at the PEP-II B Factory at SLAC. We determine the CP-odd fraction to be 0.063+/-0.055(stat)+/ 0.009(syst). The time-dependent CP asymmetry parameters Im(lambda(+)) and /lambda(+)/ are determined to be 0.05+/-0.29(stat)+/-0.10(syst) and 0.75+/ 0.19(stat)+/-0.02(syst), respectively. The standard model predicts these parameters to be -sin(2beta and 1, respectively, in the absence of penguin diagram contributions. PMID- 14525324 TI - Quantum melting of the charge-density-wave state in 1T-TiSe2. AB - We report a Raman scattering study of low-temperature, pressure-induced melting of the charge-density-wave (CDW) phase of 1T-TiSe2. Our measurements reveal that the collapse of the CDW state occurs in three stages: (i) For P<5 kbar, the pressure dependence of the CDW amplitude mode energies and intensities are indicative of a "crystalline" CDW regime; (ii) for 525 kbar, the absence of amplitude modes reveals a metallic regime in which the CDW has melted. PMID- 14525325 TI - Stripes on a 6-leg Hubbard ladder. AB - While density matrix renormalization group calculations find stripes on doped n leg t-J ladders, little is known about the possible formation of stripes on n-leg Hubbard ladders. Here we report results for a 7x6 Hubbard model with four holes. We find that a stripe forms for values of U/t ranging from 6 to 20. For U/t approximately 3-4, the system exhibits the domain wall feature of a stripe, but the hole density is very broadened. PMID- 14525326 TI - Interaction effects at crossings of spin-polarized one-dimensional subbands. AB - We report conductance measurements of ballistic one-dimensional (1D) wires defined in GaAs/AlGaAs heterostructures in an in-plane magnetic field, B. When the Zeeman energy is equal to the 1D subband energy spacing, the spin-split subband N upward arrow intersects (N+1) downward arrow, where N is the index of the spin-degenerate 1D subband. At the crossing of N=1 upward arrow and N=2 downward arrow subbands, there is a spontaneous splitting giving rise to an additional conductance structure evolving from the 1.5(2e(2)/h) plateau. With further increase in B, the structure develops into a plateau and lowers to 2e(2)/h. With increasing temperature and magnetic field the structure shows characteristics of the 0.7 structure. Our results suggest that at low densities a spontaneous spin splitting occurs whenever two 1D subbands of opposite spins cross. PMID- 14525327 TI - Enhanced shot noise in resonant tunneling via interacting localized states. AB - In a variety of mesoscopic systems shot noise is seen to be suppressed in comparison with its Poisson value. In this work we observe a considerable enhancement of shot noise in the case of resonant tunneling via localized states. We present a model of correlated transport through two localized states which provides both a qualitative and a quantitative description of this effect. PMID- 14525328 TI - Interaction effects on counting statistics and the transmission distribution. AB - We investigate the effect of weak interactions on the full counting statistics of charge transfer through an arbitrary mesoscopic conductor. We show that the main effect can be incorporated into an energy dependence of the transmission eigenvalues and study this dependence in a nonperturbative approach. An unexpected result is that all mesoscopic conductors behave at low energies such as either a single or a double tunnel junction, which divides them into two broad classes. PMID- 14525329 TI - Coherent "metallic" resistance and medium localization in a disordered one dimensional insulator. AB - It is believed that a disordered one-dimensional (1D) wire with coherent electronic conduction is an insulator with the mean resistance approximately equal e(2L/xi) and resistance dispersion Delta(rho) approximately equal e(L/xi), where L is the wire length and xi is the electron localization length. Here we show that this 1D insulator undergoes at full coherence the crossover to a 1D "metal," caused by thermal smearing and resonant tunneling. As a result, Delta(rho) is smaller than unity and tends to be L/xi independent, while grows with L/xi first nearly linearly and then polynomially, manifesting the so-called medium localization. PMID- 14525330 TI - Gapless magnetic and quasiparticle excitations due to the coexistence of antiferromagnetism and superconductivity in CeRhIn5: a study of 115In NQR under pressure. AB - We report systematic measurements of ac susceptibility, nuclear-quadrupole resonance spectrum, and nuclear-spin-lattice-relaxation time (T1) on the pressure (P)-induced heavy-fermion superconductor CeRhIn5. The temperature (T) dependence of 1/T(1) at P=1.6 GPa has revealed that antiferromagnetism (AFM) and superconductivity (SC) coexist microscopically, exhibiting the respective transition at T(N)=2.8 K and T(MF)(c)=0.9 K. It is demonstrated that SC does not yield any trace of gap opening in low-lying excitations below T(onset)(c)=2 K, but T(MF)(c)=0.9 K, followed by a T(1)T=const law. These results point to the unconventional characteristics of SC coexisting with AFM. We highlight that both of the results deserve theoretical work on the gapless nature in the low-lying excitation spectrum due to the coexistence of AFM and SC and the lack of the mean field regime below T(onset)(c)=2 K. PMID- 14525331 TI - Heat transport in proximity structures. AB - We study heat and charge transport through a normal diffusive wire coupled with a superconducting wire over the region smaller than the coherence length. Because of partial Andreev reflection of quasiparticles from the interface, the subgap thermal flow is essentially suppressed and approaches zero along with energy, which is specific for diffusive structures. Whereas the electric conductance shows a conventional reentrance effect, the thermal conductance kappa rapidly decreases with temperature which qualitatively explains the results of recent experiments. In the Andreev interferometer geometry, kappa experiences full-scale oscillations with the order parameter phase difference. PMID- 14525332 TI - Magnetic dichroism and spin structure of antiferromagnetic NiO(001) films. AB - We find that Ni L2 edge x-ray magnetic linear dichroism is fully reversed for NiO(001) films on materials with reversed lattice mismatch. We relate this phenomenon to a preferential stabilization of magnetic S domains with main spin component either in or out of the plane, via dipolar interactions. This suggests a way to selectively control spin structures in 3d systems with small spin-orbit coupling. PMID- 14525333 TI - Quenched disorder and the critical behavior of a partially frustrated system. AB - We report the direct observation of the effects of quenched disorder on the critical behavior of partially frustrated amorphous FeMnZr alloys by the systematic analysis of high-precision ac susceptibility data and dc magnetization data. Interestingly, the analysis reveals that the presence of short-range quenched disorder does not alter the actual critical behavior. However, it does affect quantities such as the Curie temperature, the peak value of effective exponent gamma, width of the peak, and crossover temperatures. The observed temperature dependence of the effective critical exponent can be understood in terms of the field-theoretical renormalization group approach. Also, the present results would help in identifying the main source of the spread in the exponent values reported in the literature. PMID- 14525334 TI - Superconducting gap for a two-leg t-J ladder. AB - Single-particle diagonal and off-diagonal Green's functions of a two-leg t-J ladder at 1/8 doping are investigated by exact diagonalizations techniques. A numerically tractable expression for the superconducting gap is proposed and the frequency dependence of the real and imaginary parts of the gap are determined. The role of the low-energy gapped spin modes, whose energies are computed by a (one-step) contractor renormalization procedure, is discussed. PMID- 14525335 TI - Micromagnetic phase transitions and spin wave excitations in a ferromagnetic stripe. AB - Magnetic excitations of micrometer-wide ferromagnetic stripes subjected to a transverse applied field have been measured between 1 and 20 GHz. The complexity of the observed response is attributed to the spatially nonuniform equilibrium spin distribution. This one is modeled analytically and numerically, which allows one to distinguish two micromagnetic phases governing the ground state. The nucleation-related phase transitions are evidenced by soft modes, while the different observed resonances are attributed to spin wave modes localized in the two phases and at their interface. PMID- 14525336 TI - Potential signature of a Kosterlitz-Thouless transition in BaNi2V2O8. AB - ESR measurements are reported for the quasi-two-dimensional honeycomb antiferromagnet BaNi2V2O8. Planar anisotropic properties are confirmed by angular dependent investigations of resonance field and linewidth. The divergence of the temperature-dependent linewidth on approaching T(N) from above is described in terms of the Kosterlitz-Thouless transition, the critical behavior close to long range magnetic order, and the 2D Heisenberg antiferromagnet. We provide arguments that the Kosterlitz-Thouless scenario is compatible with the observed critical exponent and suggest BaNi2V2O8 is a weakly anisotropic 2D Heisenberg antiferromagnet. PMID- 14525337 TI - Electrofriction and dynamic stern layers at planar charged surfaces. AB - Using dynamic simulations, the electrophoretic mobility of counterions at a substrate with fixed or mobile surface charges under the action of a lateral electric field is studied. The lateral charge inhomogeneity and corrugation of the substrate is taken into account. Because of the pronounced electrofriction between counterions and surface ions, a large fraction of counterions is practically immobilized for highly charged substrates. This explains the experimentally observed saturation of the electrophoretic mobility of charged particles in the limit of high surface charge density. PMID- 14525338 TI - Budding and tubulation in highly oblate vesicles by anchored amphiphilic molecules. AB - We studied local budding and tubulation induced in highly oblate lipid vesicles by the anchoring of either polymers having a hydrophilic backbone and grafted hydrophobic anchor groups, or by oleoyl-coenzyme A, an amphiphilic molecule important in lipid metabolism. The dynamics of bud formation, shrinkage, and readsorption is consistent with an induced spontaneous curvature coupled with local amphiphile diffusion on the membrane. We report a novel metastable state prior to bud readsorption. PMID- 14525339 TI - Possible mechanism for cold denaturation of proteins at high pressure. AB - We study cold denaturation of proteins at high pressures. Using multicanonical Monte Carlo simulations of a model protein in a water bath, we investigate the effect of water density fluctuations on protein stability. We find that above the pressure where water freezes to the dense ice phase (approximately 2 kbars) the mechanism for cold denaturation with decreasing temperature is the loss of local low-density water structure. We find our results in agreement with data of bovine pancreatic ribonuclease A. PMID- 14525340 TI - Ballistic diffusion induced by a thermal broadband noise. AB - We present a thermal broadband noise from the difference between two Ornstein Uhlenbeck noises, which can induce a ballistic diffusion, i.e., long-time mean square displacement of a free particle driven by this noise reads proportional to t(2). We apply this noise to a flashing ratchet and the mean velocity of the particle is calculated via Langevin simulation. The results show that a double peak of the mean velocity and flux reversal appears for the ratchet with large and small asymmetries, respectively; the inertia effect induces a large mean velocity and multireversal of flux. These rich and interesting phenomena are explained. PMID- 14525341 TI - Equilibrium distribution of mutators in the single fitness peak model. AB - This Letter develops an analytically tractable model for determining the equilibrium distribution of mismatch repair deficient strains in unicellular populations. The approach is based on the single fitness peak model, which has been used in Eigen's quasispecies equations in order to understand various aspects of evolutionary dynamics. As with the quasispecies model, our model for mutator-nonmutator equilibrium undergoes a phase transition in the limit of infinite sequence length. This "repair catas-trophe" occurs at a critical repair error probability of epsilon(r)=L(via)/L, where L(via) denotes the length of the genome controlling viability, while L denotes the overall length of the genome. The repair catastrophe therefore occurs when the repair error probability exceeds the fraction of deleterious mutations. Our model also gives a quantitative estimate for the equilibrium fraction of mutators in Escherichia coli. PMID- 14525342 TI - Unstable trigger waves induce various intricate dynamic regimes in a reaction diffusion system of blood clotting. AB - In this work we demonstrate that the unstable trigger waves, connecting stable and unstable spatially uniform steady states, can create intricate dynamic regimes in one-dimensional three-component reaction-diffusion model describing blood clotting. Among the most interesting regimes are the composite and replicating waves running at a constant velocity. The front part of the running composite wave remains constant, while its rear part oscillates in a complex manner. The rear part of the running replicating wave periodically gives rise to new daughter waves, which propagate in the direction opposite the parent wave. The domain of these intricate regimes in parameter space lies in the region of monostability near the region of bistability. PMID- 14525343 TI - Nanomechanical analog of a laser: amplification of mechanical oscillations by stimulated zeeman transitions. AB - We propose a magnetomechanical device that exhibits many properties of a laser. The device is formed by a nanocantilever and dynamically polarized paramagnetic nuclei of a solid sample in a strong external magnetic field. The corresponding quantum oscillator and effective two-level systems are coupled by the magnetostatic dipole-dipole interaction between a permanent magnet on the cantilever tip and the magnetic moments of the spins, so that the entire system is effectively described by the Jaynes-Cummings model. We consider the possibility of observing transient and cw lasing in this system, and show how these processes can be used to improve the sensitivity of magnetic resonance force microscopy. PMID- 14525344 TI - Preferential attachment in the protein network evolution. AB - The Saccharomyces cerevisiae protein-protein interaction map, as well as many natural and man-made networks, shares the scale-free topology. The preferential attachment model was suggested as a generic network evolution model that yields this universal topology. However, it is not clear that the model assumptions hold for the protein interaction network. Using a cross-genome comparison, we show that (a) the older a protein, the better connected it is, and (b) the number of interactions a protein gains during its evolution is proportional to its connectivity. Therefore, preferential attachment governs the protein network evolution. Evolutionary mechanisms leading to such preference and some implications are discussed. PMID- 14525345 TI - Comment on "Weak phase gamma using isospin analysis and time-dependent asymmetry in B(d)-->K(S)pi(+)pi(-)". PMID- 14525347 TI - Comment on "Scaling approach to the magnetic phase diagram of nanosized systems". PMID- 14525349 TI - Feshbach resonance in dense ultracold Fermi gases. AB - We propose a coherent framework allowing one to treat many-body effects in dense ultracold Fermi gases in the presence of a Feshbach resonance. We show that the simple effect of Pauli exclusion induces a strong modification of the basic scattering properties. In particular, this washes out the Feshbach resonance and provides a natural explanation for recent experimental findings. PMID- 14525350 TI - Diagnosis, prescription, and prognosis of a bell-state filter by quantum process tomography. AB - We apply the techniques of quantum process tomography to characterize errors and decoherence in a prototypical two-photon operation, a singlet-state filter. The quantum process tomography results indicate a large asymmetry in the process and also the required operation to correct for this asymmetry. We quantify residual errors and decoherence of the filtering operation after this modification. PMID- 14525351 TI - Quantum states and generalized observables: a simple proof of Gleason's theorem. AB - A quantum state can be understood in a loose sense as a map that assigns a value to every observable. Formalizing this characterization of states in terms of generalized probability distributions on the set of effects, we obtain a simple proof of the result, analogous to Gleason's theorem, that any quantum state is given by a density operator. As a corollary we obtain a von Neumann-type argument against noncontextual hidden variables. It follows that on an individual interpretation of quantum mechanics the values of effects are appropriately understood as propensities. PMID- 14525352 TI - Validity of the second law in nonextensive quantum thermodynamics. AB - The second law of thermodynamics in nonextensive statistical mechanics is discussed in the quantum regime. Making use of the convexity property of the generalized relative entropy associated with the Tsallis entropy indexed by q, Clausius' inequality is shown to hold in the range q in (0, 2]. This restriction on the range of the entropic index, q, is purely quantum mechanical and there exists no upper bound of q for validity of the second law in classical theory. PMID- 14525354 TI - Enhanced nonperturbative effects in jet distributions. AB - We consider the triple differential distribution d Gamma/dE(J)dm(2)(J)d Omega(J) for two-jet events at center of mass energy M, smeared over the end-point region m(2)(J)<,en-->)1H Reaction to Q2=1.45 (GeV/c)2. AB - We report new measurements of the ratio of the electric form factor to the magnetic form factor of the neutron, G(n)(E)/G(n)(M), obtained via recoil polarimetry from the quasielastic 2H(e-->,e(')n-->)1H reaction at Q2 values of 0.45, 1.13, and 1.45 (GeV/c)(2) with relative statistical uncertainties of 7.6% and 8.4% at the two higher Q2 points, which points have never been achieved in polarization measurements. PMID- 14525360 TI - Three-body recombination at large scattering lengths in an ultracold atomic gas. AB - We study three-body recombination in an optically trapped ultracold gas of cesium atoms with precise magnetic control of the s-wave scattering length a. At large positive values of a, we measure the dependence of the rate coefficient on a and confirm the theoretically predicted scaling proportional to a(4). Evidence of recombination heating indicates the formation of very weakly bound molecules in the last bound energy level. PMID- 14525361 TI - Direct measurement of transition frequencies in isolated pHe+ atoms, and new CPT violation limits on the antiproton charge and mass. AB - A radio frequency quadrupole decelerator and achromatic momentum analyzer were used to decelerate antiprotons and produce p4He+ and p3He+ atoms in ultra-low density targets, where collision-induced shifts of the atomic transition frequencies were negligible. The frequencies at near-vacuo conditions were measured by laser spectroscopy to fractional precisions of (6-19) x 10(-8). By comparing these with QED calculations and the antiproton cyclotron frequency, we set a new limit of 1 x 10(-8) on possible differences between the antiproton and proton charges and masses. PMID- 14525362 TI - Laser control of collective spontaneous emission. AB - The collective spontaneous emission of a pair of two coupled three-level radiators in vacuum is investigated in the presence of a possibly intense laser field. The parameters describing the collective interaction along with the population and decay rates of all involved dressed states are shown to be controllable by the applied laser field. In particular, all populations of the collective system may be transferred at will in a reversible way into a subradiant state, allowing effective storage and manipulation of the quantum system. PMID- 14525363 TI - Quantum entanglement of Fock states with perfectly efficient ultraslow single probe photon four-wave mixing. AB - We propose a method to achieve quantum entanglement of two Fock states with perfectly efficient, ultraslow propagation enhanced four-wave mixing. A cold atomic medium is illuminated with a two-mode cw control laser to produce coherent mixtures of excited states. An ultraslowly propagating, single-photon quantum probe field completes the four-wave mixing with 100% photon flux conversion efficiency, creating a depth dependent entanglement of two Fock states. We show that at a suitable propagation distance, a maximum entangled state is created with a single-photon wave-packet state that has 50% probability of being in each of two product-type Fock states. PMID- 14525364 TI - Red solitons: evidence of spatiotemporal instability in chi 2 spatial soliton dynamics. AB - In chi(2) three-wave mixing, the noise-seeded spatiotemporal modulational instability has a dramatic impact on the spatial soliton formation and on their stability, leading to the occurrence of a temporal breakup on the 20 fs scale and to the counterintuitive observation of spatial solitons with no apparent participation of the high-frequency field in the self-trapping. PMID- 14525365 TI - Stability and ensemble inequivalence in a globally coupled system. AB - We consider a system of globally coupled rotors, described by a set of Langevin equations, and examine the stability of the incoherent phase. The corresponding Fokker-Planck equation, providing a unified description of microcanonical and canonical ensembles, bears a few solutions, depending upon the ensemble. It is found that the stability of each solution varies differently with the temperature, revealing the inequivalence between the two ensembles. This also suggests a physical explanation of the quasistationarity observed in recent numerical results. PMID- 14525366 TI - Direct measurement of poloidal long-wavelength E x B flows in the HT-7 tokamak. AB - The poloidal long-wavelength E x B time-varying flows were directly measured using a forked Langmuir probe in the HT-7 tokamak. Low-frequency (<10 kHz) E x B flows were observed at the plasma edge, which possess many of the characteristics of zonal flows, including a poloidal long-wavelength (k(theta)rho(i) approximately 0) and narrow radial extent (k(r)rho(i) approximately 0.1). The cross bicoherence of turbulent Reynolds stress indicates the existence of nonlinear three-wave coupling processes and the generation of low-frequency E x B flows. The estimated flow-shearing rate is of the same order of magnitude as the turbulence decorrelation rate and may thus regulate the fluctuation level and thereby the turbulence-driven transport. PMID- 14525367 TI - Impact of frustrated singularities on magnetic island evolution. AB - The growth of magnetic islands is explored using the magnetohydrodynamic model in a simple slab system in which the value of the tearing mode stability parameter Delta' can be varied continuously. Unless the system is close to marginal stability reconnection is controlled by Sweet-Parker current layers, whose formation is a consequence of the inherent singular structure of magnetic island equilibria. PMID- 14525368 TI - New interpretation of alpha-particle-driven instabilities in deuterium-tritium experiments on the Tokamak Fusion Test Reactor. AB - The original description of alpha particle driven instabilities in the Tokamak Fusion Test Reactor in terms of toroidal Alfven eigenmodes (TAEs) remained inconsistent with three fundamental characteristics of the observations: (i) the variation of the mode frequency with toroidal mode number, (ii) the chirping of the mode frequency for a given toroidal mode number, and (iii) the antiballooning density perturbation of the modes. It is now shown that these characteristics can be explained by observing that cylindrical-like modes can exist in the weak magnetic shear region of the plasma that then make a transition to TAEs as the central safety factor decreases in time. PMID- 14525369 TI - Isochoric heating of solid-density matter with an ultrafast proton beam. AB - A new technique is described for the isochoric heating (i.e., heating at constant volume) of matter to high energy-density plasma states (>10(5) J/g) on a picosecond time scale (10(-12)sec). An intense, collimated, ultrashort-pulse beam of protons--generated by a high-intensity laser pulse--is used to isochorically heat a solid density material to a temperature of several eV. The duration of heating is shorter than the time scale for significant hydrodynamic expansion to occur; hence the material is heated to a solid density warm dense plasma state. Using spherically shaped laser targets, a focused proton beam is produced and used to heat a smaller volume to over 20 eV. The technique described of ultrafast proton heating provides a unique method for creating isochorically heated high energy density plasma states. PMID- 14525370 TI - Energy dissipation in gigahertz oscillators from multiwalled carbon nanotubes. AB - Using atomistic models and molecular dynamics simulations, interlayer corrugation and resistant force in a biwalled carbon nanotube are shown to be strongly dependent upon the morphology combination of the bitube. Consequently, energy dissipation in a commensurate (e.g., armchair/armchair or zigzag/zigzag) bitube oscillator is found to be much larger than that in an incommensurate (e.g., zigzag/armchair) oscillator, resulting in a decay of oscillation amplitude within a few nanoseconds in the commensurate bitube and several tens of nanoseconds in the incommensurate bitube. PMID- 14525371 TI - Optical characterization of a nanoscale incommensurate pitch in a new liquid crystal phase. AB - As a remarkable orientationally ordered soft-condensed matter system, the smectic C*(alpha) phase exhibits an incommensurate nanoscale helical pitch of more than four smectic layers. Recent theoretical advances predict the existence of a new phase showing a helical pitch of less than four smectic layers (<16 nm). By applying a novel optical technique to the study of freestanding films with increments in thickness of one smectic layer, we have measured the size of this pitch using 633 nm HeNe laser light. Our results confirm the existence of the newly predicted phase in one unique compound. PMID- 14525372 TI - Electrically deactivating nearest-neighbor donor-pair defects in Si. AB - Based on first-principles density-functional calculations, we propose a class of nearest-neighbor donor pairs that are energetically favorable in highly n-type Si. These donor pairs comprise dopant atoms either fourfold coordinated at the nearest-neighbor distance or threefold coordinated through bond-breaking relaxations. For P and As dopants, the two defect states are very close in energy, less than 0.1 eV, while the threefold coordinated state is more stable by 0.24 eV for Sb dopants. The former state has a very deep donor level close to the valence band maximum, while the defect level lies deep inside the valence band for the latter. Thus, both the donor pairs are electrically inactive at very high doping levels, and they are suggested to be responsible for the observed saturation of carriers. PMID- 14525373 TI - Multiple electron diffraction and two-dimensional crystalline order in liquid crystal thin films. AB - Electron diffraction in freestanding thin films in the crystal-B phase reveals extensive multiple diffraction in films of 14S5 but not of 4O.8, suggesting an important difference in the structural order in these films despite the similarities in their positional correlations found in earlier experiments. The result in a two-layer 14S5 film indicates surprisingly that secondary diffraction can occur in a crystal with only pseudo-long-range positional order. PMID- 14525374 TI - Evidence for a new class of defects in highly n-doped Si: donor-pair-vacancy interstitial complexes. AB - Electron channeling experiments performed on individually scanned, single columns of atoms show that in highly n-type Si grown at low temperatures the primary electrically deactivating defect cannot belong to either the widely accepted class of donor-vacancy clusters or a recently proposed class of donor pairs. First-principles calculations suggest a new class of defects consisting of two dopant donor atoms near a displaced Si atom, which forms a vacancy-interstitial pair. These complexes are consistent with the present experimental results, the measured open volume of the defects, the observed electrical activity as a function of dopant concentration, and the enhanced diffusion of impurities in the presence of deactivated dopants. PMID- 14525353 TI - Limits on D0-macro D0 mixing and CP violation from the ratio of lifetimes for decay to K-pi+, K-K+, and pi- pi+. AB - We present a measurement of D0-macro D0 mixing parameters using the ratios of lifetimes extracted from samples of D0 mesons decaying to K-pi(+), K-K+, and pi( )pi(+). Using 91 fb(-1) of data collected by the BABAR detector at the PEP-II asymmetric-energy B Factory, we obtain a value Y=[0.8+/-0.4(stat.)(+0.5)( 0.4)(syst.)]%, which, in the limit of CP conservation, corresponds to the mixing parameter y=Delta Gamma/2 Gamma. Using the difference in lifetimes of D0 and macro D0 mesons, we obtain the CP-violation parameter Delta Y=[-0.8+/ 0.6(stat.)+/-0.2(syst.)]%. PMID- 14525376 TI - Wall mediated transport in confined spaces: exact theory for low density. AB - We present a theory for the transport of molecules adsorbed in slit and cylindrical nanopores at low density, considering the axial momentum gain of molecules oscillating between diffuse wall reflections. Good agreement with molecular dynamics simulations is obtained over a wide range of pore sizes, including the regime of single-file diffusion where fluid-fluid interactions are shown to have a negligible effect on the collective transport coefficient. We show that dispersive fluid-wall interactions considerably attenuate transport compared to classical hard sphere theory. PMID- 14525377 TI - Phase transition from asymmetric to symmetric dimer structure on the Si(001) surface at high temperature. AB - The dimer configurations on the Si(001) surface at high temperatures have been investigated using the rocking curve of reflection high-energy electron diffraction. The Si(001) surface shows a displacive phase transition around 900 K, where a well-known asymmetric (tilted) dimer structure on the Si(001) at room temperature transforms to a symmetric dimer structure around 900 K. The metallic feature of the Si(001) surface above 900 K can be explained by the phase transition. PMID- 14525375 TI - Na adsorption on the Si111-(7 x 7) surface: from two-dimensional gas to nanocluster array. AB - We have systematically investigated Na adsorption on the Si(111)-(7 x 7) surface at room temperature using scanning tunneling microscopy (STM). Below the critical coverage of 0.08 monolayer, we find intriguing contrast modulation instead of localized Na adsorbates, coupled with streaky noise in the STM images, which is accompanied by monotonic work function drop. Above the critical coverage, Na clusters emerge and form a self-assembled array. Combined with first-principles theoretical simulations, we conclude that the Na atoms on the (7 x 7) surface are, while strongly bound ( approximately 2.2 eV) to the surface, highly mobile in "basins" around the Si rest atoms, forming a two-dimensional gas phase at the initial coverage, and that the cluster at the higher coverage consists of six Na atoms together with three Si adatoms. PMID- 14525378 TI - Electron localization and a confined electron gas in nanoporous inorganic electrides. AB - The nanoporous main group oxide 12CaO.7Al(2)O3 (C12A7) can be transformed from a wide-gap insulator to an electride where electrons substitute anions in cages constituting a positive frame. Our ab initio calculations of the electronic structure of this novel material give a consistent explanation of its high conductivity and optical properties. They show that the electrons confined in the inert positive frame are localized in cages and undergo hopping between neighboring cages. The results are useful for the understanding of behavior of confined electron gas of different topology and electron-phonon coupling, and for designing new transparent conductors, electron emitters, and electrides. PMID- 14525379 TI - Van der Waals density functional for layered structures. AB - To understand sparse systems, we must account for both strong local atom bonds and weak nonlocal van der Waals forces between atoms separated by empty space. A fully nonlocal functional form [Phys. Rev. B 62, 6997 (2000)]] of density functional theory (DFT) is applied here to the layered systems graphite, boron nitride, and molybdenum sulfide to compute bond lengths, binding energies, and compressibilities. These key examples show that the DFT with the generalized gradient approximation does not apply for calculating properties of sparse matter, while use of the fully nonlocal version appears to be one way to proceed. PMID- 14525380 TI - Interaction effects in conductivity of Si inversion layers at intermediate temperatures. AB - We compare the temperature dependence of resistivity rho(T) of Si-metal-oxide semiconductor field-effect transistors with the recent theory by Zala et al. In this comparison, the effective mass m* and g* factor for mobile electrons have been determined from independent measurements. An anomalous increase of rho with temperature, which has been considered as a signature of the "metallic" state, can be described quantitatively by the interaction effects in the ballistic regime. The in-plane magnetoresistance rho(B(axially)) is only qualitatively consistent with the theory; the lack of quantitative agreement indicates that the magnetoresistance is more sensitive to sample-specific effects than rho(T). PMID- 14525381 TI - Direct measurements of the spin and the cyclotron gaps in a 2D electron system in silicon. AB - Using magnetocapacitance data in tilted magnetic fields, we directly determine the chemical potential jump in a strongly correlated two-dimensional electron system in silicon when the filling factor traverses the spin and the cyclotron gaps. The data yield an effective g factor that is close to its value in bulk silicon and does not depend on the filling factor. The cyclotron splitting corresponds to the effective mass that is strongly enhanced at low electron densities. PMID- 14525382 TI - Orbital mechanisms of electron-spin manipulation by an electric field. AB - A theory of spin manipulation of quasi-two-dimensional (2D) electrons by a time dependent gate voltage applied to a quantum well is developed. The Dresselhaus and Rashba spin-orbit coupling mechanisms are shown to be rather efficient for this purpose. The spin response to a perpendicular-to-plane electric field is due to a deviation from the strict 2D limit and is controlled by the ratios of the spin, cyclotron, and confinement frequencies. The dependence of this response on the magnetic field direction is indicative of the strengths of the competing spin orbit coupling mechanisms. PMID- 14525383 TI - Design of shallow donor levels in diamond by isovalent-donor coupling. AB - Using the first-principles pseudopotential method, we have studied simultaneous isovalent and n-type doping in diamond. We show that Si induces fully occupied isovalent levels near the valence band maximum. The Si levels interact with N donor levels, making them much shallower. The donor transition energy level of the N + 4Si defect complexes is found to be 0.09 eV below the conduction band minimum, which is the shallowest level found thus far for this system. The binding energy of the N + 4Si complex is also large enough to insure its stability. PMID- 14525384 TI - Electrical transport of spin-polarized carriers in disordered ultrathin films. AB - Slow, nonexponential relaxation of electrical transport accompanied by memory effects has been induced in quench-condensed ultrathin amorphous Bi films by the application of a parallel magnetic field. This behavior, which is very similar to space-charge limited current flow, is found in extremely thin films well on the insulating side of the thickness-tuned superconductor-insulator transition. It may be the signature of a collective state that forms when the carriers are spin polarized at low temperatures and in high magnetic fields. PMID- 14525385 TI - Magnetotunneling spectroscopy of dilute Ga(AsN) quantum wells. AB - We use magnetotunneling spectroscopy to explore the admixing of the extended GaAs conduction band states with the localized N-impurity states in dilute GaAs(1 y)N(y) quantum wells. In our resonant tunneling diodes, electrons can tunnel into the N-induced E- and E+ subbands in a GaAs(1-y)N(y) quantum well layer, leading to resonant peaks in the current-voltage characteristics. By varying the magnetic field applied perpendicular to the current direction, we can tune an electron to tunnel into a given k state of the well; since the applied voltage tunes the energy, we can map out the form of the energy-momentum dispersion curves of E- and E+. The data reveal that for a small N content (approximately 0.1%) the E- and E+ subbands are highly nonparabolic and that the heavy effective mass E+ states have a significant Gamma-conduction band character even at k=0. PMID- 14525386 TI - Model for the voltage steps in the breakdown of the integer quantum Hall effect. AB - In samples used to maintain the U.S. resistance standard the breakdown of the dissipationless integer quantum Hall effect occurs as a series of dissipative voltage steps. A mechanism for this type of breakdown is proposed, based on the generation of magnetoexcitons when the quantum Hall fluid flows past an ionized impurity above a critical velocity. The calculated generation rate gives a voltage step height in good agreement with measurements on both electron and hole gases. We also compare this model to a hydrodynamic description of breakdown. PMID- 14525387 TI - Resonant tunneling of interacting electrons in a one-dimensional wire. AB - We consider the conductance of a one-dimensional wire interrupted by a double barrier structure allowing for a resonant level. Using the electron-electron interaction strength as a small parameter, we are able to build a nonperturbative analytical theory of the conductance valid in a broad region of temperatures and for a variety of the barrier parameters. We find that the conductance may have a nonmonotonic crossover dependence on temperature, specific for a resonant tunneling in an interacting electron system. PMID- 14525388 TI - Coulomb drag by small momentum transfer between quantum wires. AB - We demonstrate that in a wide range of temperatures Coulomb drag between two weakly coupled quantum wires is dominated by processes with a small interwire momentum transfer. Such processes, not accounted for in the conventional Luttinger liquid theory, cause drag only because the electron dispersion relation is not linear. The corresponding contribution to the drag resistance scales with temperature as T2 if the wires are identical, and as T5 if the wires are different. PMID- 14525389 TI - Definitive experimental evidence for two-band superconductivity in MgB2. AB - The superconducting-gap of MgB2 has been studied by high-resolution angle resolved photoemission spectroscopy. The results show that superconducting gaps with values of 5.5 and 2.2 meV open on the sigma band and the pi band, respectively, but both the gaps close at the bulk transition temperature, providing a definitive experimental evidence for the two-band superconductivity with strong interband pairing interaction in MgB2. The experiments validate the role of k-dependent electron-phonon coupling as the origin of multiple-gap superconductivity as well as the high transition temperature of MgB2. PMID- 14525390 TI - Two-step melting of the vortex solid in layered superconductors with random columnar pins. AB - We consider the melting of the vortex solid in highly anisotropic layered superconductors with a small concentration of random columnar pinning centers. Using large-scale numerical minimization of a free-energy functional, we find that melting of the low-temperature, nearly crystalline vortex solid (Bragg glass) into a vortex liquid occurs in two steps as the temperature increases: the Bragg glass and liquid phases are separated by an intermediate Bose glass phase. A suitably defined local melting temperature exhibits spatial variation similar to that observed in experiments. PMID- 14525391 TI - Resonant escape over an oscillating barrier in underdamped Josephson tunnel junctions. AB - The escape from a metastable state over an oscillating barrier of an underdamped Josephson tunnel junction has been experimentally investigated with oscillation frequency well separated from the plasma frequency of the junction. The resonant escape, namely, a minimum of the average escape time as a function of the oscillation frequency, was observed. For the oscillation frequency much smaller than the "resonant frequency," the average escape time is the average of the times required to cross over each of the barriers. On the other hand, for the oscillation frequency much greater than the "resonant frequency," the average escape time is that required to cross the average barrier. PMID- 14525392 TI - Peak Effect and the Phase Diagram of Moving Vortices in FexNi1-xZr2 Superconducting Glasses. AB - In the mixed state of type II superconductors, vortices penetrate the sample and form a correlated system due to the screening of supercurrents around them. Interestingly, we can study this correlated system as a function of density and driving force. The density, for instance, is controlled by the magnetic field B, whereas a current density j acts as a driving force F=j x B on all vortices. To minimize the pinning strength, we study a superconducting glass in which the depinning current is 10 to 1000 times smaller than in previous studies, which enables us to map out the complete phase diagram in this new regime. The diagram is obtained as a function of B, driving current, and temperature, and leads to a remarkable set of new results, which includes a huge peak effect, an additional reentrant depinning phase, and a driving force induced pinning phase. PMID- 14525393 TI - Novel dynamic ccaling regime in hole-doped La2CuO4. AB - Only 3% hole doping by Li is sufficient to suppress the long-range three dimensional (3D) antiferromagnetic order in La2CuO4. The spin dynamics of such a 2D spin liquid state at T<2 or 4-->3) needed to promote a ground state electron across the energy gap. The wavelength dependence of the damage threshold suggests that a primary mechanism for damage initiation in DKDP is a multiphoton process in which the order is reduced through excited defect state absorption. PMID- 14525400 TI - Strong energy dependence of the electron-phonon coupling strength on Bi(100). AB - We have studied the energy dependence of the electron-phonon coupling strength on Bi(100). A fit of the temperature-dependent surface state linewidth results in a change of the coupling parameter lambda from 0.20+/-0.02 to 0.72+/-0.05 as the binding energy of the surface state increases from 70 to 330 meV. This result cannot be reconciled with the usual interpretation of lambda as the electron phonon mass enhancement parameter at the Fermi energy. We suggest that this behavior is mainly caused by the strong energy dependence of the bulk density of states in this region. PMID- 14525401 TI - Essential Role of Couplings between Hearing Nonlinearities. AB - Hopf-type nonlinearities have been recently found to be the basic mechanism of the mammalian cochlear response. Physiology requires that these nonlinearities be coupled. By suitably implementing a biomorphic coupling scheme of cochlear nonlinearities, we obtain a simple cochlea model that faithfully reproduces measured basilar membrane response, validating the utility of the Hopf amplifier concept. Our results demonstrate that the correct coupling between nonlinearities is as important as the nonlinearities themselves. PMID- 14525402 TI - Two-dimensional colloid crystals obtained by coupling of flow and confinement. AB - This Letter describes the generation of 2D colloidal lattices in microchannels by coupling the laminar flow of dispersions of spherical colloids and geometrical confinement. We describe a nonequilibrium, convective, mechanism leading to formation of ordered 2D structures of both closed-packed hexagonal and non-closed packed rhombic symmetries. The number and types of possible lattices is determined by the ratio of the width of the channel to the diameter of the particle. The structures tend to return to a regular lattice after a defect is introduced; that is, for example, they tend to self-repair disorder induced by particle polydispersity, contaminants, and flow instabilities. The stability of different lattices is analyzed numerically for particles with different polydispersity. PMID- 14525403 TI - Comment on "Phase diagram of La2-xSrxCuO4 probed in the infrared: imprints of charge stripe excitations". PMID- 14525405 TI - Three-dimensional quasi-Tonks gas in a harmonic trap. AB - We analyze the macroscopic dynamics of a Bose gas in a harmonic trap with a superimposed two-dimensional optical lattice, assuming a weak coupling between different lattice sites. We consider the situation in which the local chemical potential at each lattice site can be considered as that provided by the Lieb Liniger solution. Because of the weak coupling between sites and the form of the chemical potential, the three-dimensional ground-state density profile and the excitation spectrum acquire remarkable properties different from both 1D and 3D gases. We call this system a quasi-Tonks gas. We discuss the range of applicability of this regime, as well as realistic experimental situations where it can be observed. PMID- 14525406 TI - Tkachenko modes of vortex lattices in rapidly rotating bose-einstein condensates. AB - We calculate the in-plane modes of the vortex lattice in a rotating Bose condensate, from the slowly rotating to mean-field quantum Hall limits. The Tkachenko mode frequency, linear in wave vector k for lattice rotational velocities Omega much smaller than the lowest sound wave frequency in a finite system, becomes quadratic in k in the opposite limit. The system also supports an inertial mode of frequency >or=2omega. The calculated frequencies are in excellent agreement with recent observations of Tkachenko modes by Phys. Rev. Lett., 91, 100402 (2003)]. PMID- 14525407 TI - Defect-suppressed atomic crystals in an optical lattice. AB - We present a coherent filtering scheme which dramatically reduces the site occupation number defects for atoms in an optical lattice by transferring a chosen number of atoms to a different internal state via adiabatic passage. With the addition of superlattices it is possible to engineer states with a specific number of atoms per site (atomic crystals), which are required for quantum computation and the realization of models from condensed matter physics, including doping and spatial patterns. The same techniques can be used to measure two-body spatial correlation functions. PMID- 14525408 TI - Time independent description of rapidly oscillating potentials. AB - The classical and quantum dynamics in a high frequency field are found to be described by an effective time independent Hamiltonian. It is calculated in a systematic expansion in the inverse of the frequency (omega) to order omega(-4). The work is an extension of the classical result for the Kapitza pendulum, which was calculated in the past to order omega(-2). The analysis makes use of an implementation of the method of separation of time scales and of a quantum gauge transformation in the framework of Floquet theory. The effective time independent Hamiltonian enables one to explore the dynamics in the presence of rapidly oscillating fields, in the framework of theories that were developed for systems with time independent Hamiltonians. The results are relevant, in particular, for exploring the dynamics of cold atoms. PMID- 14525409 TI - Robust long-distance entanglement and a loophole-free bell test with ions and photons. AB - Two trapped ions that are kilometers apart can be entangled by the joint detection of two photons, each coming from one of the ions, in a basis of entangled states. Such a detection is possible with linear optical elements. The use of two-photon interference allows entanglement distribution free of interferometric sensitivity to the path length of the photons. The present method of creating entangled ions also opens up the possibility of a loophole-free test of Bell's inequalities. PMID- 14525410 TI - Extension of the fluctuation theorem. AB - Heat fluctuations are studied in a dissipative system with both deterministic and stochastic components for a simple model: a Brownian particle dragged through water by a moving potential. An extension of the stationary state fluctuation theorem is derived. For infinite time, this reduces to the conventional fluctuation theorem only for small fluctuations; for large fluctuations, it gives a much larger ratio of the probabilities of the particle to absorb rather than supply heat. This persists for finite times and should be observable in experiments similar to a recent one carried out by Wang et al. PMID- 14525411 TI - Increasing the bandwidth of resonant gravitational antennas: the case of explorer. AB - Resonant gravitational wave detectors with an observation bandwidth of tens of hertz are a reality: the antenna Explorer, operated at CERN by the ROG Collaboration, has been upgraded with a new readout. In this new configuration, it exhibits an unprecedented useful bandwidth: in over 55 Hz about its center operating frequency of 919 Hz the spectral sensitivity is better than 10(-20) Hz( 1/2). We describe the detector and its sensitivity and discuss the foreseeable upgrades to even larger bandwidths. PMID- 14525412 TI - Zero-field dichroism in the solar chromosphere. AB - We explain the linear polarization of the Ca ii infrared triplet observed close to the edge of the solar disk. In particular, we demonstrate that the physical origin of the enigmatic polarizations of the 866.2 and 854.2 nm lines lies in the existence of atomic polarization in their metastable (2)D(3)(/2, 5/2) lower levels, which produces differential absorption of polarization components (dichroism). To this end, we have solved the problem of the generation and transfer of polarized radiation by taking fully into account all the relevant optical pumping mechanisms in multilevel atomic models. We argue that "zero field" dichroism may be of great diagnostic value in astrophysics. PMID- 14525413 TI - Parametric resonance in quantum field theory. AB - We present the first study of parametric resonance in quantum field theory from a complete next-to-leading order calculation in a 1/N expansion of the two-particle irreducible effective action, which includes scattering and memory effects. We present a complete numerical solution for an O(N)-symmetric scalar theory and provide an approximate analytic description of the nonlinear dynamics in the entire amplification range. We find that the classical resonant amplification at early times is followed by a collective amplification regime with explosive particle production in a broad momentum range, which is not accessible in a leading-order calculation. PMID- 14525414 TI - Hadronic density of states from string theory. AB - We present an exact calculation of the finite temperature partition function for the hadronic states corresponding to a Penrose-Guven limit of the Maldacena-Nunez embedding of the N=1 super Yang-Mills (SYM) into string theory. It is established that the theory exhibits a Hagedorn density of states. We propose a semiclassical string approximation to the finite temperature partition function for confining gauge theories admitting a supergravity dual, by performing an expansion around classical solutions characterized by temporal windings. This semiclassical approximation reveals a hadronic energy density of states of a Hagedorn type, with the coefficient determined by the gauge theory string tension as expected for confining theories. We argue that our proposal captures primarily information about states of pure N=1 SYM theory, given that this semiclassical approximation does not entail a projection onto states of large U(1) charge. PMID- 14525415 TI - Hunting for glueballs in electron-positron annihilation. AB - We calculate the cross section for the exclusive production of J(PC)=0(++) glueballs G0 in association with the J/psi in e(+)e(-) annihilation using the perturbative QCD factorization formalism. The required long-distance matrix element for the glueball is bounded by CUSB data from a search for resonances in radiative Upsilon decay. The cross section for e(+)e(-)-->J/psi+G0 at sqrt[s]=10.6 GeV is similar to exclusive charmonium-pair production e(+)e(-)- >J/psi+h for h=eta(c) and chi(c0), and is larger by a factor of 2 than that for h=eta(c)(2S). As the subprocesses gamma(*)-->(cc)(cc) and gamma(*)-->(cc)(gg) are of the same nominal order in perturbative QCD, it is possible that some portion of the anomalously large signal observed by Belle in e(+)e(-)-->J/psiX may actually be due to the production of charmonium-glueball J/psiG(J) pairs. PMID- 14525416 TI - Production mechanism of quark-gluon plasma in heavy-ion collisions. AB - A general scheme is proposed here to describe the production of semihard and soft quarks and gluons that form the bulk of the plasma in ultrarelativistic heavy-ion collisions. We show how to obtain the production rates in the extended phase space, including the color part, as a function of time in a consistent manner and without having to make ad hoc assumptions. All the required features-the back reaction on QCD vacuum, the non-Markovian nature of the production, and the quasi particle nature of the partons-are naturally incorporated. We illustrate the results with a realistic albeit toy model and also show how physically tenable source terms may be obtained. PMID- 14525417 TI - Nucleon-nucleon coincidence spectra in the nonmesonic weak decay of lambda hypernuclei and the gamma(n)/gamma(p) puzzle. AB - By combining a one-meson-exchange model for the LambdaN-->nN transition in finite nuclei with an intranuclear cascade code, we have obtained nucleon-nucleon (angular and energy) coincidence distributions from the nonmesonic weak decay of (5)(Lambda)He and (12)(Lambda)C hypernuclei. Although, due to the elimination of interferences, two-nucleon coincidences are expected to give a cleaner extraction (with respect to single nucleon observables) of the ratio Gamma(n)/Gamma(p) identical with Gamma(Lambdan-->nn)/Gamma(Lambdap-->np), we show that the effect of the final state interactions is still important even when applying favorable energy and angular cuts. The agreement of our results for the ratio N(nn)/N(np) between the number of nn and np emitted pairs with preliminary KEK coincidence data allows us to conclude that Gamma(n)/Gamma(p) for (5)(Lambda)He should be small and close to the value of 0.46 predicted by our meson-exchange model. PMID- 14525418 TI - Shannon entropy as an indicator of atomic avoided crossings in strong parallel magnetic and electric fields. AB - Avoided crossings are the most distinctive atomic spectroscopic features in the presence of magnetic and electric fields. We point out the role of Shannon's information entropy as an indicator or predictor of these phenomena by studying the dynamics of some excited states of hydrogen in the presence of parallel magnetic and electric fields. Moreover, in addition to the well-known energy level repulsion, it is found that Shannon's entropy manifests the informational exchange of the involved states as the magnetic field strength is varied across the narrow region where an avoided crossing occurs. PMID- 14525419 TI - Rescattering of ultralow-energy electrons for single ionization of Ne in the tunneling regime. AB - Electron emission for single ionization of Ne by 25 fs, 1.0 PW/cm(2) laser pulses at 800 nm has been investigated in a kinematically complete experiment using a "reaction microscope." Mapping the complete final state momentum space with high resolution, a distinct local minimum is observed at P(e parallel )=0, where P(e parallel ) is the electron momentum parallel to the laser polarization. Whereas tunneling theory predicts a maximum at zero momentum, our findings are in good agreement with recent semiclassical predictions which were interpreted to be due to "recollision." PMID- 14525420 TI - Hyperfine polarization and its normal gradient coefficient of (87)Rb atoms in the vicinity (approximately 10(-5) cm) of coated and uncoated Pyrex glass surfaces. AB - We have made regionally specific measurement of the hyperfine polarization of 87Rb atoms in the vicinity ( approximately 10(-5) cm) of coated and uncoated Pyrex glass surfaces. We find that the polarization near an uncoated surface decreases rapidly with decreasing distance from the surface whereas for a silicone-coated surface the polarization is independent of the distance from the surface. We have also determined the normal gradient coefficient micro (S.I) of the hyperfine polarization in uncoated cells. In a representative uncoated cell, at a Rb density 7.35 x 10(13) cm(-3) and a pump beam intensity 1.3 W/cm(2), we find micro(S.I)=24+/-7 microm(-1). PMID- 14525421 TI - Bethe logarithm and QED shift for lithium. AB - A novel finite basis set method is used to calculate the Bethe logarithm for the ground 2 (2)S(1/2) and excited 3 (2)S(1/2) states of lithium. The basis sets are constructed to span a huge range of distance scales within a single calculation, leading to well-converged values for the Bethe logarithm. The results are used to calculate an accurate value for the complete quantum electrodynamic energy shift up to order alpha(3) Ry. The calculated 3 (2)S(1/2)-2 (2)S(1/2) transition frequency for 7Li is 27 206.092 6(9) cm(-1), and the ionization potential for the 2 (2)S(1/2) state is 43 487.158 3(6) cm(-1). The 7Li-6Li isotope shift is also considered, and all the results compared with experiment. PMID- 14525422 TI - Two-loop Bethe-logarithm correction in hydrogenlike atoms. AB - We calculate the two-loop Bethe logarithm correction to atomic energy levels in hydrogenlike systems. The two-loop Bethe logarithm is a low-energy quantum electrodynamic (QED) effect involving multiple summations over virtual excited atomic states. Although much smaller in absolute magnitude than the well-known one-loop Bethe logarithm, the two-loop analog is quite significant when compared to the current experimental accuracy of the 1S-2S transition: It contributes 8.19 and -0.84 kHz for the 1S and the 2S state, respectively. The two-loop Bethe logarithm has been the largest unknown correction to the hydrogen Lamb shift to date. Together with the ongoing measurement of the proton charge radius at the Paul Scherrer Institute, its calculation will bring theoretical and experimental accuracy for the Lamb shift in atomic hydrogen to the level of 10(-7). PMID- 14525423 TI - Photoelectron spectroscopy of C(84) dianions. AB - We report the laser photoelectron spectra of doubly negatively charged C84 (D2 and D(2d)) using 532 nm and 355 nm radiation. From these spectra, values for the second electron affinity and vertical detachment energy, as well as upper and lower limits for the repulsive Coulomb barrier, are obtained. These values are discussed in the context of classical electrostatic models. The experimental spectra are compared with the accessible excited states of the C-84 product ion calculated in the framework of time dependent density functional theory. PMID- 14525424 TI - Low-energy electron diffraction and resonances in DNA and other helical macromolecules. AB - We propose a framework to calculate the intermolecular multiple elastic scattering of low-energy electrons from helical macromolecules and indicate how it affects the resonant capture cross section. Using a model of DNA, an appreciable enhancement of the elastic and resonant capture cross sections is predicted at incident energies below 15 eV. These results may qualitatively explain the observed prominence of low-energy resonances in strand breaking of plasmid DNA. PMID- 14525425 TI - Electron impact ionization of sodium Rydberg atoms below 2 eV. AB - We have observed electron impact ionization of highly excited sodium Rydberg atoms in ns and nd states, n=35-51, below E=2 eV electron kinetic energy with energy resolution 0.25 eV. Measured absolute cross sections near 0 eV range from sigma(35d) approximately 7 x 10(-10) to sigma(50d) approximately 4 x 10(-9) cm(2). The energy dependence is consistent with that of widely used binary encounter approximation cross sections, and sigma(n) follows a power law in n. The measured cross sections are 14 to 24 times larger than theoretically predicted values. This enhancement may signal the effect of large polarizabilities of high Rydberg states not yet accounted for in ionization theories. PMID- 14525426 TI - Multiband vector lattice solitons. AB - We predict multiband vector solitons in nonlinear periodic systems, using photonic lattices as a prime example. The solitons consist of two optical fields arising from different bands of the transmission spectrum, which involve both bound state and radiation mode components. PMID- 14525427 TI - Multigap discrete vector solitons. AB - We analyze nonlinear collective effects in periodic systems with multigap transmission spectra such as light in waveguide arrays or Bose-Einstein condensates in optical lattices. We reveal that the interband interactions in nonlinear periodic structures can be efficiently managed by controlling their geometry. We predict novel types of discrete vector solitons supported by nonlinear coupling between different band gaps and study their stability. PMID- 14525428 TI - Manipulation and removal of defects in spontaneous optical patterns. AB - Defects play an important role in a number of fields dealing with ordered structures. We demonstrate theoretically and experimentally the possibility of an active manipulation of defects in terms of an externally induced motion. We focus on the spontaneous formation of two-dimensional spatial structures in a nonlinear optical system, a liquid crystal light valve under single optical feedback. For a particular parameter setting, a spontaneously formed hexagonal intensity pattern contains several dislocation-type defects. A scheme based on Fourier filtering allows us to restore spatial order in a selectable part of the pattern. Starting without control, the controlled area is progressively expanded, such that defects are swept out of the pattern. PMID- 14525429 TI - Decay kinetic properties of atoms in photonic crystals with absolute gaps. AB - Decay kinetic properties of a two-level atom near the band edges of photonic crystals (PCs) with absolute gaps are studied based on the Green's function expression for the evolution operator. The local coupling strength between the photons and an atom is evaluated by an exact numerical method. It is found that the decay behavior of an excited atom can be fundamentally changed by the variation of the atomic position: Weisskopf-Wigner and non-Weisskopf-Wigner decay phenomena occur at different atomic positions in the PCs as a result of a significant difference in the local coupling strength. Our finding implies that it is possible to engineer the luminescence spectrum by controlling the atomic position. PMID- 14525430 TI - Synchronization of active mechanical oscillators by an inertial load. AB - Motivated by the operation of myogenic (self-oscillatory) insect flight muscle, we study a model consisting of a large number of identical oscillatory contractile elements joined in a chain, whose end is attached to a damped mass spring oscillator. When the inertial load is small, the serial coupling favors an antisynchronous state in which the extension of one oscillator is compensated by the contraction of another, in order to preserve the total length. However, a sufficiently massive load can synchronize the oscillators and can even induce oscillation in situations where isolated elements would be stable. The system has a complex phase diagram displaying quiescent, synchronous and antisynchronous phases, as well as an unusual asynchronous phase in which the total length of the chain oscillates at a different frequency from the individual active elements. PMID- 14525431 TI - Localized transverse bursts in inclined layer convection. AB - We investigate a novel bursting state in inclined layer thermal convection in which convection rolls exhibit intermittent, localized, transverse bursts. With increasing temperature difference, the bursts increase in duration and number while exhibiting a characteristic wave number, magnitude, and size. We propose a mechanism which describes the duration of the observed bursting intervals and compare our results to bursting processes in other systems. PMID- 14525432 TI - Phenomenology of Rayleigh-Taylor turbulence. AB - I analyze the advanced mixing regime of the Rayleigh-Taylor incompressible turbulence in the small Atwood number Boussinesq approximation. The prime focus of my phenomenological approach is to resolve the temporal behavior and the small scale spatial correlations of velocity and temperature fields inside the mixing zone, which grows as proportional, variant t(2). I show that the "5/3"-Kolmogorov scenario for velocity and temperature spectra is realized in three spatial dimensions with the viscous and dissipative scales decreasing in time, proportional, variant t(-1/4). The Bolgiano-Obukhov scenario is shown to be valid in two dimensions with the viscous and dissipative scales growing, proportional, variant t(1/8). PMID- 14525433 TI - Torque on an ideal plasma. AB - Ripples in the confining field may exert a torque on a rotating plasma. Time reversal symmetry implies that this torque should vanish for an ideal plasma. However, even in an apparently ideal plasma, singularities can give rise to a nonzero torque. This torque is evaluated for a simple configuration. Although the primary force is magnetic, an essential contribution arises from other nonlinear terms in the equations of motion. The net force is confined to the singular layer, in the direction of the ripple wave vector and related to the energy absorbed in the layer. PMID- 14525434 TI - Elastic stiffness of the nuclear-spin system in tetragonal U2D2 nuclear-ordered solid (3)He. AB - We have measured temperature dependences of sound velocity for both longitudinal and transverse sound in nuclear-ordered U2D2 solid 3He with several crystal orientations along the melting curve. The sound velocity change was proportional to T4 for all sound modes and crystal orientations and was attributed to the nuclear-spin part of the internal energy. We extracted six-independent elastic stiffness of the nuclear-spin part and obtained Gruneisen constants of the spin wave velocity for four-independent strains. Gruneisen constants for compressional strain were larger than those for shear strain. Using the multiple-spin-exchange model, we explain the anisotropy of Gruneisen constants in tetragonal symmetry. PMID- 14525435 TI - Giant axial electrostrictive deformation in carbon nanotubes. AB - An exceptionally large axial electrostrictive deformation is demonstrated in single walled carbon nanotubes using Hartree-Fock and density functional quantum mechanics simulations. Armchair and zigzag open-ended tubes and capped tubes are studied and in all of them the external electric field induced axial strains can be greater than 10% for a field strength within 1 V/A. The corresponding volumetric and gravimetric work capacities are predicted to be three and six orders higher than those of the best known ferroelectric, electrostrictive, magnetostrictive materials and elastomers, respectively. PMID- 14525436 TI - Three-particle correlations in simple liquids. AB - We use videomicroscopy to follow the phase-space trajectory of a two-dimensional colloidal model liquid and calculate three-point correlation functions from the measured particle configurations. Approaching the fluid-solid transition by increasing the strength of the pair-interaction potential, one observes the gradual formation of a crystal-like local order due to triplet correlations, while being still deep inside the fluid phase. Furthermore, we show that in a strongly interacting system the Born-Green equation can be satisfied only with the full triplet correlation function but not with three-body distribution functions obtained from superposing pair correlations (Kirkwood superposition approximation). PMID- 14525437 TI - Sliding friction with polymer brushes. AB - Using high-resolution shear force measurements, we examine in detail the frictional drag between rubbing surfaces bearing end-tethered polymeric surfactants (brushes). The drag attains a maximum on initial motion, attributed to elastic stretching of the chains, which falls by a cascade of relaxations to a value characteristic of kinetic friction. This has a very weak velocity dependence, attributed to chain moieties dragging within a self-regulating, mutual interpenetration zone. When sliding stops, the shear stress across the polymer layers decays logarithmically with time, consistent with the relaxation of a network of dangling ends. PMID- 14525438 TI - Potential-energy landscape study of the amorphous-amorphous transformation in H(2)O. AB - We study the potential energy landscape explored during a compression decompression cycle for the simple point charge extended model of water. During the cycle, the system changes from low density amorphous (LDA) ice to high density amorphous ice. After the cycle, the system does not return to the same region of the landscape, supporting the interesting possibility that more than one significantly different configuration corresponds to LDA. We find that the regions of the landscape explored during this transition have properties remarkably different from those explored in thermal equilibrium in the liquid phase. PMID- 14525439 TI - Glass formation criterion for various glass-forming systems. AB - A conceptual approach to evaluate glass-forming ability for various glass-forming systems has been proposed from a physical metallurgy point of view. It was found that the glass-forming ability for noncrystalline materials was related mainly to two factors, i.e., 1/(T(g)+T(l)) and Tx (wherein Tx is the onset crystallization temperature, T ( g) the glass transition temperature, and T(l) the liquidus temperature), and could be predicated by a unified parameter gamma defined as T(x)/(T(g)+T(l)). This approach was confirmed and validated by experimental data in various glass-forming systems including oxide glasses, cryoprotectants, and metallic glasses. PMID- 14525440 TI - Stroboscopic X-ray imaging of vibrating dislocations excited by 0.58 GHz phonons. AB - We report on direct imaging, by means of stroboscopic x-ray topography, of phonon induced dislocation vibrations. X-ray images taken from LiNbO3 crystals excited by 0.58 GHz surface acoustic waves, showed individual acoustic wave fronts as well as their distortions when crossing the dislocation line. The observed contrast is well explained by considering the dynamic deformation field of vibrating dislocation. Comparing simulated deformation maps and x-ray images permitted determination of the local velocities of vibrating dislocations and their viscosity coefficients. We found unexpectedly high velocity values (not far from the speed of sound) and extremely low viscosity coefficients, 2-3 orders of magnitude lower than previously measured in ductile materials. PMID- 14525441 TI - Does the arrhenius temperature dependence of the Johari-Goldstein relaxation persist above T(g)? AB - Dielectric spectra of the polyalcohols sorbitol and xylitol were measured under isobaric pressures up to 1.8 GPa. At elevated pressure, the separation between the alpha and beta relaxation peaks is larger than at ambient pressure, enabling the beta relaxation times to be unambiguously determined. Taking advantage of this, we show that the Arrhenius temperature dependence of the beta relaxation time does not persist for temperatures above T(g). This result, consistent with inferences drawn from dielectric relaxation measurements at ambient pressure, is obtained directly, without the usual problematic deconvolution the beta and alpha processes. PMID- 14525442 TI - "Magic" heteroepitaxial growth on vicinal surfaces. AB - The step period (Lambda) of vicinal surfaces can be used as a new parameter for the control of metallic heteroepitaxial growth. This is evidenced here in the case of Ag/Cu(211). The deposition of 1 monolayer (ML) exhibits a c(2 x 10) superstructure leading to the formation of [111] steps in the Ag adlayer in contrast with the original [100] steps for the Cu substrate. This wetting layer can be viewed as a (133) Ag plane and it will be the starting point for the epitaxial growth. The deposition of 4 ML shows that the thin Ag film results homogeneous and no twins or stacking faults are detected. Moreover, the film grows along the [133] axis which is the orientation that minimizes the misfit between Cu(211) and the Ag film. Thus, the use of a regular stepped substrate allows one to select the crystallographic orientation of the growth and seems to be a way to avoid the creation of stacking faults. PMID- 14525443 TI - (2 x 1)-(1 x 1) phase transition on Ge(001): dimer breakup and surface roughening. AB - Using low energy electron microscopy, we have investigated the (2 x 1)-(1 x 1) phase transition occurring above 925 K on Ge(001). Dimer breakup has been identified as the physical origin of this transition. A quantitative description of the dimer concentration during the transition involves configuration entropy of random monomers within the dimer matrix. The dimer formation energy amounts to 1.2+/-0.3 eV. Dimer breakup promotes reversible surface disorder by step proliferation and irreversible surface roughening above 1130 K. PMID- 14525444 TI - Unusual thermodynamic properties and nonergodicity in ferroelectric superlattices. AB - The properties of [Pb(Zr(1-x(1))Ti(x(1)))O(3)](n)/[Pb(Zr(1-x(2))Ti(x(2)))O(3)](n) superlattices, with a 2n period, are simulated using an ab initio based approach. The x(1) and x(2) compositions are chosen to be located across the morphotropic phase boundary of the corresponding disordered alloys, while the (x(1)+x(2))/2 average composition lies inside this boundary. These superlattices exhibit an unusual thermodynamic phase transition sequence, including a triclinic ground state. They also have the kind of peculiar free-energy landscape yielding nonergodicity. The effects responsible for these anomalies are discussed. PMID- 14525445 TI - Hubbard model on decorated lattices. AB - We introduce a family of lattices for which the Hubbard model and its natural extensions can be quasiexactly solved, i.e., solved for the ground and low energy states. In particular, we show rigorously that the ground state of the Hubbard model with off-site Coulomb repulsions on a decorated Kagome lattice is an ordered array of local currents. The low energy theory describing this chiral state is an S=1 / 2 XY model, where each spin degree of freedom represents the two possible chiralities of each local current. PMID- 14525446 TI - Coherent backscattering near the two-dimensional metal-insulator transition. AB - We have studied corrections to conductivity due to the coherent backscattering in low-disordered two-dimensional electron systems in silicon for a range of electron densities including the vicinity of the metal-insulator transition, where the dramatic increase of the spin susceptibility has been observed earlier. We show that the corrections, which exist deeper in the metallic phase, weaken upon approaching the transition and practically vanish at the critical density, thus suggesting that the localization is suppressed near and at the transition even in zero field. PMID- 14525447 TI - Optical probing of a fractionally charged quasihole in an incompressible liquid. AB - In photoluminescence spectroscopy of a low-mobility two-dimensional electron gas subjected to a quantizing magnetic field, we observe an anomaly around nu=1 / 3 at a very low temperature (0.1 K) and an intermediate electron density (0.9 x 10(11) cm(-2)). The anomaly is explained as due to perturbation of the incompressible liquid at the Laughlin state due to close proximity of a localized charged exciton which creates a fractionally charged quasihole in the liquid. The anomaly of approximately 2 meV can be destroyed by applying a small thermal energy of approximately 0.2 meV that is enough to close the quasihole energy gap. PMID- 14525448 TI - Direct experimental evidence for the Ruderman-Kittel-Kasuya-Yosida interaction in rare-earth metals. AB - We show that the ferromagnetic heavy rare-earth (RE) metals show a transport spin polarization at the Fermi level in the majority spin, whereas in ferromagnetic light rare earths it is in the minority spin. The sign of the polarization is in agreement with what is expected due to the Ruderman-Kittel-Kasuya-Yosida (RKKY) coupling formalism. We show that magnetotransport measurements on magnetic multilayer samples containing magnetic REs provide a unique opportunity to verify the RKKY coupling scheme in pure rare-earth metals, allowing us to probe both the sign and temperature dependence of the spin-density oscillation. PMID- 14525449 TI - Coherent versus sequential electron tunneling in quantum dots. AB - Manifestations of quantum coherence in the electronic conductance through nearly closed quantum dots in the Coulomb-blockade regime are addressed. We show that quantum coherent tunneling processes explain some puzzling statistical features of the conductance peak heights observed in recent experiments at low temperatures. We employ the constant interaction model and the random matrix theory to model the quantum dot electronic interactions and its single-particle statistical fluctuations, taking full account of the finite decay width of the quantum dot levels. PMID- 14525450 TI - Phase diagram for quantum hall bilayers at nu=1. AB - We present a phase diagram for a double quantum well bilayer electron gas in the quantum Hall regime at a total filling factor nu=1, based on exact numerical calculations of the topological Chern number matrix and the (interlayer) superfluid density. We find three phases: a quantized Hall state with pseudospin superfluidity, a quantized Hall state with pseudospin "gauge-glass" order, and a decoupled composite Fermi liquid. Comparison with experiments provides a consistent explanation of the observed quantum Hall plateau, Hall drag plateau, and vanishing Hall drag resistance, as well as the zero-bias conductance peak effect, and suggests some interesting points to pursue experimentally. PMID- 14525451 TI - Addition energies of fullerenes and carbon nanotubes as quantum dots: the role of symmetry. AB - Using density-functional theory calculations, we investigate the addition energy (AE) of quantum dots formed of fullerenes or closed single-wall carbon nanotubes. We focus on the connection between symmetry and oscillations in the AE spectrum. In the highly symmetric fullerenes the oscillation period is large because of the large level degeneracy and Hund's rule. For long nanotubes, the AE oscillation is fourfold. Adding defects destroys the spatial symmetry of the tubes, leaving only spin degeneracy; correspondingly, the fourfold behavior is destroyed, leaving an even/odd behavior which is quite robust. We use our symmetry results to explain recent experiments. PMID- 14525452 TI - Metamorphosis of the quantum Hall ferromagnet at nu=2/5. AB - We report on the dramatic evolution of the quantum Hall ferromagnet in the fractional quantum Hall regime at nu=2/5 filling. A large enhancement in the characteristic time scale gives rise to a dynamical transition into a novel quantized Hall state. The observed Hall state is determined to be a zero temperature phase distinct from the spin-polarized and spin-unpolarized nu=2/5 fractional quantum Hall states. It is characterized by a strong temperature dependence and puzzling correlation between temperature and time. PMID- 14525453 TI - Mott gap excitations and resonant inelastic x-ray scattering in doped cuprates. AB - Predictions are made for the momentum- and carrier-dependent degradation of the Mott gap upon doping in high-T(c) cuprates as would be observed in Cu K-edge resonant inelastic x-ray scattering (RIXS). The two-dimensional Hubbard model with second- and third-nearest-neighbor hopping terms has been studied by numerical exact diagonalization. Special emphasis is placed on the particle-hole asymmetry of the Mott gap excitations. We argue that the Mott gap excitations observed by RIXS are significantly influenced by the interaction between charge carriers and antiferromagnetic correlations. PMID- 14525454 TI - Specific heat of classical disordered elastic systems. AB - We study the thermodynamics of disordered elastic systems, applied to vortex lattices in the Bragg glass phase. Using the replica variational method we compute the specific heat of pinned vortons in the classical limit. We find that the contribution of disorder is positive, linear at low temperature, and exhibits a maximum. It is found to be important compared to other contributions, e.g., core electrons, mean field, and nonlinear elasticity that we evaluate. The contribution of droplets is subdominant at weak disorder in d=3. PMID- 14525455 TI - Universal spin-flip transition in itinerant antiferromagnets. AB - We reveal a universal spin-flip (SF) transition as a function of temperature in spin-density-wave (SDW) systems. At low temperatures the antiferromagnetic (AFM) polarization is parallel to the applied field and above a critical temperature the AFM polarization flips perpendicular to the field. This transition occurs in any SDW system and may be considered as a qualitative probe of the itinerant character of AFM in a given material. Our SF transition may provide an explanation to the long-standing puzzle of the SF transition observed in chromium and may be at the origin of the equally puzzling SDW-I to SDW-II transition in Bechgaard salts for which we make experimental predictions. PMID- 14525456 TI - Locally accessible information: how much can the parties gain by cooperating? AB - We investigate measurements of bipartite ensembles restricted to local operations and classical communication and find a universal Holevo-like upper bound on the locally accessible information. We analyze our bound and exhibit a class of states which saturate it. Finally, we link the bound to the problem of quantification of the nonlocality of the operations necessary to extract locally inaccessible information. PMID- 14525457 TI - Effects of charge fluctuation on two-membrane instability and fusion. AB - Membrane fusion is fundamental to diverse biological processes ranging from intercellular and intracellular transport to egg fertilization. We study the effects of coupling between membrane undulation and charge fluctuation on its fusion. We find that, at concentrations of millimolar range, multivalent cations such as calcium in solution induce a strong correlated-charge fluctuation on each membrane, leading to inversion and overcondensation of surface charges. When the charge fluctuation is cooperatively coupled to undulation, two apposing membranes undergo a dynamic instability to spontaneous growth of in-phase undulation with submicron wavelengths, thereby greatly reducing fusion barrier. PMID- 14525458 TI - Adiabatic pumping mechanism for ion motive ATPases. AB - An ion motive ATPase is a membrane protein that pumps ions across the membrane at the expense of the chemical energy of adenosine triphosphate (ATP) hydrolysis. Here we describe how an external electric field, by inducing transitions between several protein configurations, can also power this pump. The underlying mechanism may be very similar to that of a recently constructed adiabatic electron pump [Science 283, 1905 (1999)]]. PMID- 14525459 TI - Mechanisms of molecular manipulation with the scanning tunneling microscope at room temperature: chlorobenzene/si(111)-(7 x 7). AB - We report a systematic experimental investigation of the mechanism of desorption of chlorobenzene molecules from the Si(111)-(7 x 7) surface induced by the STM at room temperature. We measure the desorption probability as a function of both tunneling current and a wide range of sample bias voltages between -3 V and +4 V. The results exclude field desorption, thermally induced desorption, and mechanical tip-surface effects. They indicate that desorption is driven by the population of negative (or positive) ion resonances of the chemisorbed molecule by the tunneling electrons (or holes). Density functional calculations suggest that these resonant states are associated with the pi orbitals of the benzene ring. PMID- 14525460 TI - Mechanism for rapid self-assembly of block copolymer nanoparticles. AB - Amphiphilic block copolymers in solution spontaneously self-assemble when the solvent quality for one block is selectively decreased. We demonstrate that, for supersaturation ratio changes [d(S)/dt] over 10(5) per second from equilibrium, nanoparticles are obtained with a formation mechanism and size dependent on the jumping rate and magnitude. The threshold rate for homogeneous precipitation is determined by the induction time of a particle, equivalent to the diffusion limited fusion of copolymer chains to form a corona of overlapping soluble brushes. Via determination of the induction time with a novel confined impinging jets mixer and use of a scaling relation, the interfacial free energy of a block copolymer nanoparticle was measured for the first time. PMID- 14525461 TI - Public channel cryptography by synchronization of neural networks and chaotic maps. AB - Two different kinds of synchronization have been applied to cryptography: synchronization of chaotic maps by one common external signal and synchronization of neural networks by mutual learning. By combining these two mechanisms, where the external signal to the chaotic maps is synchronized by the nets, we construct a hybrid network which allows a secure generation of secret encryption keys over a public channel. The security with respect to attacks, recently proposed by Shamir et al., is increased by chaotic synchronization. PMID- 14525462 TI - Comment on "scaling of the interface roughness in Fe-Cr superlattices: self affine versus non-self-affine". PMID- 14525464 TI - Intermittently decreased beat-to-beat variability in congestive heart failure. PMID- 14525466 TI - Phase conjugation of a quantum-degenerate atomic fermion beam. AB - We discuss the possibility of phase conjugation of an atomic Fermi field via nonlinear wave mixing in an ultracold gas. It is shown that for a beam of fermions incident on an atomic phase-conjugate mirror, a time-reversed backward propagating fermionic beam is generated similar to the case in nonlinear optics. By adopting an operational definition of the phase, we show that it is possible to infer the presence of the phase-conjugate field by the loss of the interference pattern in an atomic interferometer. PMID- 14525467 TI - Observation of Tkachenko oscillations in rapidly rotating Bose-Einstein condensates. AB - We directly image Tkachenko waves in a vortex lattice in a dilute-gas Bose Einstein condensate. The low (sub-Hz) resonant frequencies are a consequence of the small but nonvanishing elastic shear modulus of the vortex-filled superfluid. The frequencies are measured for rotation rates as high as 98% of the centrifugal limit for the harmonically confined gas. Agreement with a hydrodynamic theory worsens with increasing rotation rate, perhaps due to the increasing fraction of the volume displaced by the vortex cores. We also observe two low-lying m=0 longitudinal modes at about 20 times higher frequency. PMID- 14525468 TI - Observation of geometric phases for mixed states using NMR interferometry. AB - Examples of geometric phases abound in many areas of physics. They offer both fundamental insights into many physical phenomena and lead to interesting practical implementations. One of them, as indicated recently, might be an inherently fault-tolerant quantum computation. This, however, requires one to deal with geometric phases in the presence of noise and interactions between different physical subsystems. Despite the wealth of literature on the subject of geometric phases very little is known about this very important case. Here we report the first experimental study of geometric phases for mixed quantum states. We show how different they are from the well-understood, noiseless, pure-state case. PMID- 14525469 TI - Constraining hadronic superfluidity with neutron star precession. AB - I show that the standard picture of the neutron star core containing coexisting neutron and proton superfluids, with the proton component forming a type II superconductor threaded by flux tubes, is inconsistent with observations of long period (approximately 1 yr) precession in isolated pulsars. I conclude that either the two superfluids coexist nowhere in the stellar core, or the core is a type I superconductor rather than type II. Either possibility would have interesting implications for neutron star cooling and theories of spin jumps (glitches). PMID- 14525470 TI - Renormalization-group evolution of the B-meson light-cone distribution amplitude. AB - An integro-differential equation governing the evolution of the leading-order B meson light-cone distribution amplitude is derived. The anomalous dimension in this equation contains a logarithm of the renormalization scale, whose coefficient is identified with the cusp anomalous dimension of Wilson loops. The exact solution of the evolution equation is obtained, from which the asymptotic behavior of the distribution amplitude is derived. These results can be used to resum Sudakov logarithms entering the hard-scattering kernels in QCD factorization theorems for exclusive B decays. PMID- 14525471 TI - An infrared renormalization group limit cycle in QCD. AB - We use effective field theories to show that small increases in the up and down quark masses would move QCD very close to the critical renormalization group trajectory for an infrared limit cycle in the three-nucleon system. We conjecture that QCD can be tuned to the critical trajectory by adjusting the quark masses independently. At the critical values of the quark masses, the binding energies of the deuteron and its spin-singlet partner would be tuned to zero and the triton would have infinitely many excited states with an accumulation point at the 3-nucleon threshold. The ratio of the binding energies of successive states would approach a universal constant that is close to 515. PMID- 14525472 TI - Proton-number fluctuation as a signal of the QCD critical end point. AB - We argue that the event-by-event fluctuation of the proton number is a meaningful and promising observable for the purpose of detecting the QCD critical end point in heavy-ion collision experiments. The long range fluctuation of the order parameter induces a characteristic correlation between protons which can be measured. The proton fluctuation also manifests itself as anomalous enhancement of charge fluctuations near the end point, which might be already seen in existing data. PMID- 14525473 TI - Dynamics of the pi(-)p-->pi(0)pi(0)n reaction for p(pi(-))<750 MeV/c. AB - Data are presented for the reaction pi(-)p-->pi(0)pi(0)n in the range from threshold to p(pi(-))=750 MeV/c. The systematics of the data and multipole analyses are examined for sensitivity to a f(0)(600) ("sigma") meson. A one-pion exchange mechanism is found to be very weak, or absent. The reaction appears to become dominated by sequential pi(0) decays through the Delta(1232) resonance as the beam momentum increases, along with substantial interference effects from several competing mechanisms. PMID- 14525474 TI - Low-energy coexisting band in 154Gd. AB - A low-energy coexisting band Jpi (Ex keV) 0(+) (1182), 2(+) (1418), 4(+) (1701) is identified in the deformed nucleus, 154Gd. Detailed gamma-ray spectroscopy following the beta decays of 154Eu (J=3), (g,m(1),m(2))154Tb (J=0,3,7) is used to establish this structure. The structure is explained in terms of the pairing and deformation degrees of freedom, a "pairing isomer," which results from the nu[505] upward arrow Nilsson intruder orbital. PMID- 14525475 TI - How to measure the spreading width for the decay of superdeformed nuclei. AB - A new expression for the branching ratio for the decay via the E1 process in the normal-deformed band of superdeformed nuclei is given within a simple two-level model. Using this expression, the spreading or tunneling width gamma (downward arrow) for superdeformed decay can be expressed entirely in terms of experimentally known quantities. We show how to determine the tunneling matrix element V from the measured value of gamma (downward arrow) and a statistical model of the energy levels. The accuracy of the two-level approximation is verified by considering the effects of the other normal-deformed states. PMID- 14525476 TI - Wave packet movements near the conical intersection between two excited potential surfaces may create observable molecular oscillations. AB - A femtosecond pump-probe experiment is performed on tetrakis(dimethylamino)ethylene. The evolution resulting from the pi-pi(*) excitation of the CC double bond corresponds to movement along a single adiabatic potential surface with deformation along several coordinates and passage near a conical intersection. Surprisingly, this movement excites the umbrella mode of the amino groups, resulting in a measurable oscillation regime. PMID- 14525477 TI - Polarization squeezing with cold atoms. AB - We study the interaction of a nearly resonant linearly polarized laser beam with a cloud of cold cesium atoms in a high finesse optical cavity. We show theoretically and experimentally that the cross-Kerr effect due to the saturation of the optical transition produces quadrature squeezing on both the mean field and the orthogonally polarized vacuum mode. An interpretation of this vacuum squeezing as polarization squeezing is given and a method for measuring quantum Stokes parameters for weak beams via a local oscillator is developed. PMID- 14525478 TI - Free-space propagation of diffuse light: theory and experiments. AB - In this Letter we present a simple and novel theoretical approach for modeling the intensity distribution from an arbitrarily shaped turbid volume in a noncontact geometry by considering diffuse light propagation in free space. This theory is validated with experiments for a diffusive volume of known geometry in a noncontact situation, both with and without the presence of an embedded absorber. The implications of this new formulation in the context of optical tomography in turbid media are discussed. PMID- 14525479 TI - Frequency-resolved optical gating of femtosecond pulses in the extreme ultraviolet. AB - Femtosecond extreme ultraviolet (XUV) pulses were fully characterized for the first time by using a newly developed cross-correlation frequency-resolved optical gating (FROG) technique in the XUV region. This method utilizes laser assisted two-photon ionization as a nonlinear optical process. Near-infrared pulses characterized by FROG were used as a reference. The amplitude and phase of XUV pulses with a pulse duration of 10 fs were found to be in good agreement with a model analysis, taking into account phase modulation by ionization, self-phase modulation, and the atomic dipole phase. PMID- 14525480 TI - Morphology and scaling of impact craters in granular media. AB - We present the results of experiments on impact craters formed by dropping a steel ball vertically into a container of small glass beads. As the energy of impact increases, we observe a progression of crater morphologies analogous to that seen in craters on the moon. We find that both the diameter and the depth of the craters are proportional to the 1/4 power of the energy. The ratio of crater diameter to rim-to-floor depth is constant for low-energy impacts, but increases at higher energy, similar to what is observed for lunar craters. PMID- 14525481 TI - Granular medium under vertical tapping: change of compaction and convection dynamics around the liftoff threshold. AB - A granular medium submitted to vertical tapping reveals simultaneously compaction and convection. The two phenomena are directly coupled and their dynamics can be quantified by a characteristic compaction time and by an estimation of the convective downhill speed along the wall. A remarkable change of behavior is observed around the liftoff acceleration threshold of the whole packing, with a drastic slowing down of both dynamics below this threshold. Above it, a collective shock wave densifies the packing at each tap, whereas, below it, cumulative localized rearrangements will compact the entire system in the long time range. PMID- 14525482 TI - Entrainment of a spatially extended nonlinear structure under selective forcing. AB - The response of a nonlinear state to a variable forcing periodic in space is studied in an extended dynamical system consisting of a liquid crystal layer driven to convection. Both the statics and the dynamics of the entrainment and the locking effects are analyzed. The dynamics of the evolution are controlled by topological singularities that allow a diffusion of the phase. The mechanisms involved are related to the role of the defects in systems undergoing spontaneous symmetry breakings. PMID- 14525483 TI - Anatomy of a bathtub vortex. AB - We present experiments and theory for the "bathtub vortex," which forms when a fluid drains out of a rotating cylindrical container through a small drain hole. The fast down-flow is found to be confined to a narrow and rapidly rotating "drainpipe" from the free surface down to the drain hole. Surrounding this drainpipe is a region with slow upward flow generated by the Ekman layer at the bottom of the container. This flow structure leads us to a theoretical model similar to one obtained earlier by Lundgren [J. Fluid Mech. 155, 381 (1985)]], but here including surface tension and Ekman upwelling, comparing favorably with our measurements. At the tip of the needlelike surface depression, we observe a bubble-forming instability at high rotation rates. PMID- 14525484 TI - Evidence of ultrashort electron bunches in laser-plasma interactions at relativistic intensities. AB - The second harmonic of the laser light (2omega(0)) is observed on the rear side of thick solid targets irradiated by a laser beam at relativistic intensities. This emission is explained by the acceleration by the laser pulse in front of the target of short bunches of electrons separated by the period (or half the period) of the laser light. When reaching the rear side of the target, these electron bunches emit coherent transition radiation at 2omega(0). The observations indicate that, in our conditions, the minimum fraction of the laser energy transferred to these electron bunches is of the order of 1%. PMID- 14525485 TI - Instability of the 2S electron bubbles. AB - The 2S electron bubble placed in liquid helium has been previously believed to be spherical. We show that the 2S bubble is morphologically unstable at pressures above -1.23 bars. The 2S state being known to be radially unstable at pressures below -1.33 bars, the result leaves only a very narrow pressure range in which it can be found in a spherical configuration. Our stability analysis indicates that the 2S bubble is unstable against perturbations proportional to any of the third spherical harmonics Y(3m). Our numerical simulations show that there exist nonspherical stable configurations, such as the ones Maris and Konstantinov predicted for the 1P, 1D, and 2P electron bubbles and confirmed experimentally for the 1P. We believe that the 2S bubbles can also be produced and that our prediction will yield itself to experimental verification. PMID- 14525486 TI - Probing vortices in 4He nanodroplets. AB - We present static and dynamical properties of linear vortices in 4He droplets obtained from density functional calculations. By comparing the adsorption properties of different atomic impurities embedded in pure droplets and in droplets where a quantized vortex has been created, we suggest that Ca atoms should be the dopant of choice to detect vortices by means of spectroscopic experiments. PMID- 14525487 TI - Thermal conductivity of liquid 3He in aerogel: a gapless superfluid. AB - We have measured the thermal conductivity of liquid 3He in 98% aerogel at ultralow temperatures. Aerogel introduces disorder on a scale comparable to the superfluid coherence length. At low pressures the liquid in the aerogel shows normal-state behavior with conductivity linear in temperature. At pressures above approximately 6 bars the onset of superfluidity suppresses the conductivity and the thermal conductivity again tends towards linear behavior in the very low temperature limit, providing strong evidence that here the liquid 3He in the aerogel is behaving as a gapless superfluid. PMID- 14525488 TI - Field-induced structural aging in glasses at ultralow temperatures. AB - In nonequilibrium experiments on the glasses Mylar and BK7, we measured the excess dielectric response after the temporary application of a strong electric bias field at millikelvin temperatures. A model recently developed describes the observed long time decays qualitatively for Mylar [Phys. Rev. Lett. 90, 105501(2003)]], but fails for BK7. In contrast, our results on both samples can be described by including an additional mechanism to the mentioned model with temperature independent decay times of the excess dielectric response. As the origin of this novel process beyond the "tunneling model" we suggest bias field induced structural rearrangements of "tunneling states" that decay by quantum mechanical tunneling. PMID- 14525489 TI - Chemically active substitutional nitrogen impurity in carbon nanotubes. AB - We investigate the nitrogen substitutional impurity in semiconducting zigzag and metallic armchair single-wall carbon nanotubes using ab initio density functional theory. At low concentrations (less than 1 at. %), the defect state in a semiconducting tube becomes spatially localized and develops a flat energy level in the band gap. Such a localized state makes the impurity site chemically and electronically active. We find that if two neighboring tubes have their impurities facing one another, an intertube covalent bond forms. This finding opens an intriguing possibility for tunnel junctions, as well as the functionalization of suitably doped carbon nanotubes by selectively forming chemical bonds with ligands at the impurity site. If the intertube bond density is high enough, a highly packed bundle of interlinked single-wall nanotubes can form. PMID- 14525490 TI - Atomic-resolution electron energy loss spectroscopy imaging in aberration corrected scanning transmission electron microscopy. AB - The "delocalization" of inelastic scattering is an important issue for the ultimate spatial resolution of innershell spectroscopy in the electron microscope. It is demonstrated in a nonlocal model for electron energy loss spectroscopy (EELS) that delocalization of scanning transmission electron microscopy (STEM) images for single, isolated atoms is primarily determined by the width of the probe, even for light atoms. We present experimental data and theoretical simulations for Ti L-shell EELS in a [100] SrTiO3 crystal showing that, in this case, delocalization is not significantly increased by dynamical propagation. Issues relating to the use of aberration correctors in the STEM geometry are discussed. PMID- 14525491 TI - Effect of pressure relaxation on the mechanisms of short-pulse laser melting. AB - The kinetics and microscopic mechanisms of laser melting of a thin metal film are investigated in a computational study that combines molecular dynamics simulations with a continuum description of the laser excitation and subsequent relaxation of the conduction band electrons. Two competing melting mechanisms, homogeneous nucleation of liquid regions inside the crystalline material and propagation of melting fronts from external surfaces, are found to be strongly affected by the dynamics of the relaxation of the laser-induced pressure. PMID- 14525492 TI - Mechanisms and rates of interstitial H2 diffusion in crystalline C60. AB - Parallel replica dynamics and minimum energy path calculations have been used to study the diffusion mechanisms of H2 in fcc C60. Isolated interstitial H2 molecules bind preferentially in the lattice octahedral (O) sites and diffuse by hopping between O and tetrahedral sites. The simulations reveal an unexpected mechanism involving an H2 molecule diffusing through an already occupied O site, creating an H2 dimer, with a lower activation barrier than diffusion into an empty O site. Kinetic Monte Carlo simulations of a lattice model based on these mechanisms indicate that events involving dimers greatly enhance the self diffusion rates of interstitial H2 in fcc C60. PMID- 14525493 TI - Resonant photon tunneling enhancement of the van der Waals friction. AB - We study the van der Waals friction between two flat metal surfaces in relative motion. For good conductors, we find that normal relative motion gives a much larger friction than for parallel relative motion. The friction may increase by many orders of magnitude when the surfaces are covered by adsorbates, or can support low-frequency surface plasmons. In this case, the friction is determined by resonant photon tunneling between adsorbate vibrational modes, or surface plasmon modes. PMID- 14525494 TI - Higher surface energy of free nanoparticles. AB - We present an accurate online method for the study of size-dependent evaporation of free nanoparticles allowing us to detect a size change of 0.1 nm. This method is applied to Ag nanoparticles. The linear relation between the onset temperature of evaporation and the inverse of the particle size verifies the Kelvin effect and predicts a surface energy of 7.2 J/m(2) for free Ag nanoparticles. The surface energy of nanoparticles is significantly higher as compared to that of the bulk and is essential for processes such as melting, coalescence, evaporation, growth, etc., of nanoparticles. PMID- 14525495 TI - Layer ordering and faulting in (GaAs)n/(AlAs)n ultrashort-period superlattices. AB - We report studies of (GaAs)(n)/(AlAs)(n) ultrashort-period superlattices using synchrotron x-ray scattering. In particular, we demonstrate that interfaces of these superlattices contain features on two different length scales: namely, random atomic mixture and ordered mesoscopic domains. Both features are asymmetric on the two interfaces (AlAs-on-GaAs and GaAs-on-AlAs) for n>2. Periodic compositional stacking faults, arising from the intrinsic nature of molecular-beam epitaxy, are found in the superlattices. In addition, the effect of growth interruption on the interfacial structure is discussed. The relevant scattering theory is developed to give excellent fits to the data. PMID- 14525496 TI - Quantum charge fluctuations and the polarizability of the single-electron box. AB - We measure the average charge on the island of a single-electron box, with an accuracy of two thousandths of an electron. Thermal fluctuations alone cannot account for the dependence of the average charge on temperature, on external potential, or on the quasiparticle density of states in the metal from which the box is formed. In contrast, we find excellent agreement between these measurements and a theory that treats the quantum fluctuations of charge perturbatively. PMID- 14525497 TI - Quantum coherence of image-potential states. AB - The quantum dynamics of the two-dimensional image-potential states in front of the Cu(100) surface is measured by scanning tunneling microscopy and spectroscopy. The dispersion relation and the momentum resolved phase-relaxation time of the first image-potential state are determined from the quantum interference patterns in the local density of states at step edges. It is demonstrated that the tip-induced Stark shift does not affect the motion of the electrons parallel to the surface. PMID- 14525498 TI - Spin liquid state in an organic Mott insulator with a triangular lattice. AB - 1H NMR and static susceptibility measurements have been performed in an organic Mott insulator with a nearly isotropic triangular lattice, kappa-(BEDT TTF)2Cu2(CN)(3), which is a model system of frustrated quantum spins. The static susceptibility is described by the spin S=1/2 antiferromagnetic triangular lattice Heisenberg model with the exchange constant J approximately 250 K. Regardless of the large magnetic interactions, the 1H NMR spectra show no indication of long-range magnetic ordering down to 32 mK, which is 4 orders of magnitude smaller than J. These results suggest that a quantum spin liquid state is realized in the close proximity of the superconducting state appearing under pressure. PMID- 14525499 TI - Driven depinning of strongly disordered media and anisotropic mean-field limits. AB - Extended systems driven through strong disorder are modeled generically using coarse-grained degrees of freedom that interact elastically in the directions parallel to the drive and slip along at least one of the directions transverse to the motion. In the limit of infinite-range elastic and viscous coupling this model has a tricritical point separating a region where the depinning is continuous, in the universality class of elastic depinning, from a region where depinning is hysteretic. Many of the collective transport models discussed in the literature are special cases of the generic model. PMID- 14525500 TI - Quantum-classical reentrant relaxation crossover in Dy2Ti2O7 spin ice. AB - We have studied spin relaxation in the spin ice compound Dy2Ti2O7 through measurements of the ac magnetic susceptibility. While the characteristic spin relaxation time (tau) is thermally activated at high temperatures, it becomes almost temperature independent below T(cross) approximately 13 K. This behavior, combined with nonmonotonic magnetic field dependence of tau, indicates that quantum tunneling dominates the relaxational process below that temperature. As the low-entropy spin ice state develops below T(ice) approximately 4 K, tau increases sharply with decreasing temperature, suggesting the emergence of a collective degree of freedom for which thermal relaxation processes again become important as the spins become strongly correlated. PMID- 14525501 TI - In situ photoemission study of the room temperature ferromagnet ZnGeP2:Mn. AB - The chemical states of the ZnGeP(2):Mn interface which shows ferromagnetism above room temperature have been studied by photoemission spectroscopy. Mn deposition on the ZnGeP2 substrate heated to 400 degrees C induced Mn substitution for Zn and then the formation of metallic Mn-Ge-P compounds. Depth profile studies have shown that Mn 3d electrons changed their character from itinerant to localized along the depth, and in the deep region, dilute divalent Mn species (<5% Mn) was observed with a coexisting metallic Fermi edge of non-Mn 3d character. The possibility of hole doping through Mn substitution for Ge and/or Zn vacancy is discussed. PMID- 14525502 TI - Wave-vector-dependent exciton exchange interaction. AB - The exchange interaction for the yellow 1S orthoexciton in Cu2O is derived up to the order K2. The resulting exchange splittings are verified experimentally using high resolution spectroscopy. In agreement with theory the fine structure shows a characteristic dependence on the direction of the wave vector. PMID- 14525503 TI - Coherent control of polariton parametric scattering in semiconductor microcavities. AB - In a pump-probe experiment, we have been able to control, with phase-locked probe pulses, the ultrafast nonlinear optical emission of a semiconductor microcavity, arising from polariton parametric amplification. This evidences the coherence of the polariton population near k=0, even for delays much longer than the pulse width. The control of a large population at k=0 is possible although the probe pulses are much weaker than the large polarization they control. With rising pump power the dynamics of the scattering get faster. Just above threshold the parametric scattering process shows unexpected long coherence times, whereas when pump power is risen the contrast decays due to a significant pump reservoir depletion. The weak pulses at normal incidence control the whole angular emission pattern of the microcavity. PMID- 14525504 TI - Midinfrared optical breakdown in transparent dielectrics. AB - Optical breakdown measurements for transparent dielectrics are reported for 1 ps laser pulses as a function of mid-IR wavelength from 4.7 to 7.8 microm. For wide gap dielectrics seed electrons are generated by tunnel ionization with subsequent avalanche ionization and laser absorption by dense plasma. For narrow-gap dielectrics tunnel ionization alone leads to dense plasma formation. PMID- 14525505 TI - Origin of the "waterfall" effect in phonon dispersion of relaxor perovskites. AB - We have undertaken an inelastic neutron scattering study of the perovskite relaxor ferroelectric Pb(Zn(1/3)Nb(2/3))O3 with 8% PbTiO3 (PZN-8%PT) in order to elucidate the origin of the previously reported unusual kink on the low frequency transverse phonon dispersion curve (known as the "waterfall effect"). We show that its position (q(wf)) depends on the choice of the Brillouin zone and that the relation of q(wf) to the size of the polar nanoregions is highly improbable. The waterfall phenomenon is explained in the framework of a simple model of coupled damped harmonic oscillators representing the acoustic and optic phonon branches. PMID- 14525506 TI - Entanglement of formation for symmetric Gaussian states. AB - We show that for a fixed amount of entanglement, two-mode squeezed states are those that maximize Einstein-Podolsky-Rosen-like correlations. We use this fact to determine the entanglement of formation for all symmetric Gaussian states corresponding to two modes. This is the first instance in which this measure has been determined for genuine continuous variable systems. PMID- 14525507 TI - Quantum computation with a one-dimensional optical lattice. AB - We present an economical dynamical control scheme to perform quantum computation on a one-dimensional optical lattice, where each atom encodes one qubit. The model is based on atom tunneling transitions between neighboring sites of the lattice. They can be activated by external laser beams resulting in a two-qubit phase gate or in an exchange interaction. A realization of the Toffoli gate is presented, which requires only a single laser pulse and no individual atom addressing. PMID- 14525508 TI - Multiparticle entanglement purification for graph states. AB - We introduce a class of multiparticle entanglement purification protocols that allow us to distill a large class of entangled states. These include cluster states, Greenberger-Horne-Zeilinger states, and various error correction codes all of which belong to the class of two-colorable graph states. We analyze these schemes under realistic conditions and observe that they are scalable; i.e., the threshold value for imperfect local operations does not depend on the number of parties for many of these states. When compared to schemes based on bipartite entanglement purification, the protocol is more efficient and the achievable quality of the purified states is larger. As an application we discuss an experimental realization of the protocol in optical lattices which allows one to purify cluster states. PMID- 14525509 TI - Sequence dependent DNA-mediated conduction. AB - We report on coherent charge transport studies in periodic Poly(dG)-Poly(dC) and aperiodic lambda-phage DNA sequences. The extent and efficiency of charge transfer is discussed as a function of sequence dependent energetics, of temperature dependent base-base couplings, and in relation with experiments. PMID- 14525510 TI - Deformation of cross-linked semiflexible polymer networks. AB - Networks of filamentous proteins play a crucial role in cell mechanics. These cytoskeletal networks, together with various cross-linking and other associated proteins largely determine the (visco)elastic response of cells. In this Letter we study a model system of cross-linked, stiff filaments in order to explore the connection between the microstructure under strain and the macroscopic response of cytoskeletal networks. We find two distinct regimes as a function primarily of cross-link density and filament rigidity: one characterized by affine deformation and one by nonaffine deformation. We characterize the crossover between these two. PMID- 14525511 TI - Elasticity of stiff polymer networks. AB - We study the elasticity of a two-dimensional random network of rigid rods ("Mikado model"). The essential features incorporated into the model are the anisotropic elasticity of the rods and the random geometry of the network. We show that there are three distinct scaling regimes, characterized by two distinct length scales on the elastic backbone. In addition to a critical rigidity percolation region and a homogeneously elastic regime we find a novel intermediate scaling regime, where the elasticity is dominated by bending deformations. PMID- 14525512 TI - Membranes with rotating motors. AB - We study collections of rotatory motors confined to two-dimensional manifolds. These systems show a nontrivial collective behavior since the rotational motion leads to a repulsive hydrodynamic interaction between motors. While for high rotation speed motors might exhibit crystalline order, they form at low speed a disordered phase where diffusion is enhanced by velocity fluctuations. These effects should be experimentally observable for motors driven by external fields and for dipolar biological motors embedded into lipid membranes in a viscoelastic solvent. PMID- 14525513 TI - Small polarons in dry DNA. AB - We report ab initio calculations for positively charged fragments of dry poly(dC) poly(dG) DNA, with up to 4 C-G pairs. We find a strong hole-lattice coupling and clear evidence for the formation of small polarons. The largest geometry distortions occur in only one or two base pairs. They involve the stretching of weak bonds within each base pair, increasing the distance of positive hydrogens, and decreasing that of negative oxygens, to the region in which the hole localizes. We obtain an energy of approximately 0.30 eV for the polaron formation, nearly independent of the chain size. From it, we can estimate an activation energy for polaron hopping of approximately 0.15 eV, consistent with the available experimental value. PMID- 14525514 TI - Activated bond-breaking processes preempt the observation of a sharp glass-glass transition in dense short-ranged attractive colloids. AB - We study-using molecular dynamics simulations-the temperature dependence of the dynamics in a dense short-ranged attractive colloidal glass to find evidence of the kinetic glass-glass transition predicted by the ideal mode coupling theory. According to the theory, the two distinct glasses are stabilized, one by excluded volume and the other by short-ranged attractive interactions. By studying the density autocorrelation functions, we discover that the short-ranged attractive glass is unstable. Indeed, activated bond-breaking processes slowly convert the attractive glass into the hard-sphere one, preempting the observation of a sharp glass-glass transition. PMID- 14525515 TI - Doppler effect of nonlinear waves and superspirals in oscillatory media. AB - Nonlinear waves emitted from a moving source are studied. A meandering spiral in a reaction-diffusion medium provides an example in which waves originate from a source exhibiting a back-and-forth movement in a radial direction. The periodic motion of the source induces a Doppler effect that causes a modulation in wavelength and amplitude of the waves ("superspiral"). Using direct simulations as well as numerical nonlinear analysis within the complex Ginzburg-Landau equation, we show that waves subject to a convective Eckhaus instability can exhibit monotonic growth or decay as well as saturation of these modulations depending on the perturbation frequency. Our findings elucidate recent experimental observations concerning superspirals and their decay to spatiotemporal chaos. PMID- 14525516 TI - Periodic droplet formation in chemically patterned microchannels. AB - Simulations show that, when a phase-separated binary AB fluid is driven to flow past chemically patterned substrates in a microchannel, the fluid exhibits unique morphological instabilities. For the pattern studied, these instabilities give rise to the simultaneous, periodic formation of monodisperse droplets of A in B and B in A. The system bifurcates between time-independent behavior and different types of regular, nondecaying oscillations in the structural characteristics. The surprisingly complex behavior is observed even in the absence of hydrodynamic interactions and arises from the interplay between the fluid flow and patterned substrate, which introduces nonlinearity into the dynamical system. PMID- 14525517 TI - Structure of extremely nanosized and confined In-O species in ordered porous materials. AB - Perturbed-angular correlation, x-ray absorption, and small-angle x-ray scattering spectroscopies were suitably combined to elucidate the local structure of highly diluted and dispersed InOx species confined in the porous of the ZSM5 zeolite. This novel approach allow us to determined the structure of extremely nanosized In-O species exchanged inside the 10-atom-ring channel of the zeolite, and to quantify the amount of In2O3 crystallites deposited onto the external zeolite surface. PMID- 14525518 TI - Tidal effects on gravity experiments with a balance. PMID- 14525520 TI - Comment on "Universality of small scale turbulence". PMID- 14525521 TI - Comment on "Identification of lattice vacancies on the two sublattices of SiC". PMID- 14525523 TI - The "ping-pong" protocol can be attacked without eavesdropping. PMID- 14525524 TI - Extracellular matrix of the developing ovarian follicle. AB - Ovaries can be considered tissues in which endocrine organs--follicles and corpora lutea--continually grow and regress. Follicles have both epithelial and stromal layers in which cell migration or movement, cell division, specialization and differentiation, and death occur. A fluid-filled antrum develops and at ovulation the epithelial cells undergo an epithelial to mesenchymal transition into luteal cells. Although growth factors and hormones are very important in some of these processes, the extracellular matrix participates in all of them. Importantly, the matrix is diverse in composition and cells rarely behave without reference to the composition and structure of the matrix. When follicles commence growing, the follicular basal lamina changes in its composition from containing all six alpha chains of collagen type IV to only alpha 1 and alpha 2. Perlecan and nidogen 1 subsequently become components of the follicular basal lamina, and there is an increase in the amount of laminin chains alpha 1, beta 2 and gamma 1, at least in cows. Late in follicular development and on atresia some follicles contain laminin alpha 2. On atresia the follicular basal lamina is not degraded as occurs at ovulation, but can be breached by cells from the thecal layer if granulosa cells no longer align it. Other matrix components are present and also change during follicular development. Versican was identified in all the follicular layers and has been found to play a key role together with inter-alpha trypsin inhibitor, tumour necrosis factor alpha-stimulated gene 6 (TSG-6) and hyaluronan in cumulus oocyte expansion and fertility. Recent studies are directed at investigating the regulation of the matrix and its function in the ovary. PMID- 14525525 TI - Glucocorticoid metabolism and reproduction: a tale of two enzymes. AB - Within potential target cells, the actions of physiological glucocorticoids (cortisol and corticosterone) are modulated by isoforms of the enzyme 11 beta hydroxysteroid dehydrogenase (11 beta HSD). To date, two isoforms of 11 beta HSD have been cloned: 11 beta HSD1 acts predominantly as an NADP(H)-dependent reductase to generate active cortisol or corticosterone, and 11 beta HSD2 is a high affinity NAD(+)-dependent enzyme that catalyses the enzymatic inactivation of glucocorticoids. Whereas the regeneration of active glucocorticoids by 11 beta HSD1 has been implicated in the cellular mechanisms of pituitary function, ovulation and parturition, the enzymatic inactivation of cortisol and corticosterone by 11 beta HSD enzymes appears to be central to the protection of gonadal steroidogenesis, prevention of intra-uterine growth retardation, and lactation. Recent evidence indicates that follicular fluid contains endogenous modulators of cortisol metabolism by 11 beta HSD1, the concentrations of which are associated with the clinical outcome of assisted conception cycles and are altered in cystic ovarian disease. In conclusion, the two cloned isoforms of 11 beta HSD fulfil diverse roles in a wide range of reproductive processes from conception to lactation. PMID- 14525526 TI - Functionality of the spindle checkpoint during the first meiotic division of mammalian oocytes. AB - The spindle checkpoint ensures accurate chromosome segregation by delaying anaphase until all chromosomes are correctly aligned on the microtubule spindle. Although this mechanism is conserved throughout eukaryotic evolution, it is unclear whether it operates during meiosis in female mammals. The results of the present study show that in mouse oocytes spindle alterations prevent both chromosome segregation and MPF (M phase promoting factor) inactivation during the first meiotic M phase. Moreover, the spindle checkpoint component budding uninhibited by benzimidazole 1 (BUB1) localizes to kinetochores and is phosphorylated until anaphase of both meiotic M phases. Both localization and phosphorylation are similar to those observed in oocytes at microtubule depolymerization. In addition, the kinetochore localization and phosphorylation of BUB1 do not depend on the MOS/.../MAPK pathway. These data indicate that the spindle checkpoint is probably active during meiotic maturation in mouse oocytes. BUB1 remains associated with kinetochores and is phosphorylated during the metaphase arrest of the second meiotic M phase, indicating that this protein may also play a role in the natural metaphase II arrest in mammalian oocytes. PMID- 14525527 TI - Pregnancy-induced changes in substance P and neurokinin 1 receptor (NK1-R) expression in the rat uterus. AB - Adrenergic nerve fibres of the mammalian uterus degenerate during pregnancy. The behaviour of peptidergic fibres, such as substance P-positive fibres and of its preferred neurokinin 1 receptor (NK1-R), is poorly studied in the pregnant rat uterus. The present study analysed the changes in substance P immunoreactivity and in the expression of NK1-R protein in the uterus of non-pregnant, pregnant (days 7, 14 and 21) and postpartum rats (days 1, 8 and 22) by immunohistology, dot blot analysis and western blot analysis. In non-pregnant rats, substance P positive fibres were localized to the myometrium; these fibres progressively disappeared during gestation and were almost absent at term (day 21). At day 22 post partum, substance P-positive fibres had recovered to numbers comparable with those in the non-pregnant uterus. Dot blot analysis revealed a significant decrease in the immunoreactivity of substance P in the uterus at mid-pregnancy (day 14) and especially at term. Expression of the NK1-R protein showed a progressive increase throughout pregnancy reaching a peak on day 1 post partum; downregulation of NK1-R protein occurred on day 8 post partum. The low and high expressions of NK1-R protein were coincident with a large number of eosinophils and almost no eosinophils in the uterus at oestrus and at term, respectively. It was concluded that substance P immunoreactivity is inversely correlated with NK1 R protein expression in the pregnant and postpartum uterus. The marked upregulation of NK1-R protein at term and after birth indicates that the NK1-R may be involved in the complex regulation of labour and postpartum physiology. However, it is likely that the NK1-protein is not involved in the recruitment of eosinophils into the uterus at oestrus. PMID- 14525528 TI - Effects of in vivo administration of epidermal growth factor (EGF) on uterine contractility, prostaglandin production and timing of parturition in rats. AB - Prostaglandins synthesized by cyclooxygenases elicit uterine contractions during labour. Nitric oxide synthases (NOS) produce nitric oxide (NO), which maintains uterine quiescence during pregnancy. Epidermal growth factor (EGF) interacts with prostaglandins and NO in many biological systems. The aim of this work was to study the effect of the in vivo administration of EGF on uterine contractility, prostaglandin production and timing of parturition in rats. EGF was injected into the uterine lumen of pregnant rats on day 20, 21 or 22 of gestation. Intra uterine administration of 500 ng EGF on day 21 of gestation delayed parturition for 18 h compared with control rats. Administration of EGF was able to: (i) reduce cyclooxygenase expression in the uterus (determined by western blot analysis) and production of prostaglandins by the uterus (evaluated by conversion of [(14)C]arachidonate to labelled prostaglandins); (ii) decrease prostaglandin concentrations in amniotic fluid (radioimmunoassay); (iii) increase NO production (evaluated by conversion of [(14)C]arginine into [(14)C]citrulline); (iv) increase serum progesterone concentrations to more than control concentrations (P<0.05; radioimmunoassay); and (v) reduce the amplitude of the uterine contractions. The overall effect was a delay in the onset of delivery. This in vivo effect raises the question of whether exogenous EGF plays a role in the initiation of parturition. PMID- 14525529 TI - Proliferation and apoptosis in bovine placentomes during pregnancy and around induced and spontaneous parturition as well as in cows retaining the fetal membranes. AB - Placental growth can be achieved by either cellular proliferation or hypertrophy. Tissue regeneration and the nutrition of the fetus via embryotrophe require high rates of cellular turnover and the so-called pre-term 'maturation' of the placenta is correlated with a reduction of maternal crypt epithelial cells. Placentomes of 45 pregnant cows were collected from an abattoir to assess the role of proliferation and apoptosis in placental physiology and pathology. Placentomes were also taken from five cows undergoing premature Caesarean section and from ten naturally calving cows immediately after the expulsion of the fetus. Five of these animals had not released the fetal membranes after 12 h. Tissue sections of placentome were assessed for the Ki-67 protein; the TUNEL procedure was performed and verified by transmission electron microscopy. The maternal crypt epithelium and the fetal chorionic epithelium had a higher percentage of Ki 67-positive cells than the stroma. The percentage of Ki-67-positive cells increased significantly during pregnancy in fetal chorionic epithelium and was significantly decreased in fetal chorionic epithelium and maternal crypt epithelium after the expulsion of the fetus in comparison with tissue from month 9 of pregnancy. The number of apoptotic cells increased significantly during pregnancy in maternal crypt epithelium, maternal stroma and fetal chorionic epithelium as detected in slaughtered animals. Significantly more apoptotic fetal chorionic epithelial cells were found in animals retaining their fetal membranes in comparison with prepartum cattle during month 9 of pregnancy, at premature section and in animals releasing the fetal membranes completely. The results strongly indicate that bovine placentomes have cell type-specific rates of cellular turnover reflecting tissue growth, embryotrophe and placental maturation. Retention of fetal membranes is characterized by a large number of fetal chorionic epithelial cells undergoing apoptosis immediately after the expulsion of the fetus. This finding indicates that incomplete maturation of placentomes plays an important role in fetal membrane retention and that massive apoptosis after the expulsion of the fetus should be the consequence of diminished blood supply to the uterus, as verified in a recent study. PMID- 14525530 TI - Influence of progesterone supplementation during the first third of pregnancy on fetal and placental growth in overnourished adolescent ewes. AB - Overnourishing adolescent ewes throughout pregnancy promotes maternal tissue synthesis at the expense of placental growth, which in turn leads to a major decrease in lamb birth weight. As maternal dietary intakes are inversely related to peripheral progesterone concentrations in these adolescent dams, it was hypothesized that sup-optimal progesterone concentrations in overnourished dams may compromise the growth of the differentiating conceptus resulting in fewer uterine caruncles being occupied and, hence, fewer placentomes formed. This hypothesis was tested by supplementing overnourished adolescent dams with exogenous progesterone during early pregnancy and determining the impact on pregnancy outcome at term. Embryos recovered from superovulated adult ewes inseminated by a single sire were transferred in singleton to the uterus of peripubertal adolescent recipients. After transfer of embryos, ewes were offered a moderate or high amount of a complete diet (n=11 per group). A further high intake group received a progesterone supplement each day from day 5 to day 55 of gestation (term=145 days) to restore circulating progesterone concentrations to moderate values throughout the first third of pregnancy (n=11). For ewes establishing pregnancies (n=7 per group), live weight gain during the first 100 days of gestation was 66+/-4, 323+/-17 and 300+/-7 g per day, body condition score at term was 2.1+/-0.05, 3.0+/-0.08 and 3.1+/-0.07 units and the duration of gestation after spontaneous delivery was 148+/-1.7, 144+/-0.8 and 143+/-0.8 days for the moderate intake, high intake and high intake plus progesterone groups, respectively. At delivery, fetal cotyledon mass (136+/-12.1 versus 57+/-8.2g, P<0.001) and lamb birth weight (5164+/-151 versus 2893+/-381 g, P<0.001) were higher in moderate intake than in high intake dams. Progesterone supplementation restored circulating concentrations to moderate values during the first third of gestation. Lamb birth weight in the high intake plus progesterone group (4150+/ 389 g) was intermediate between the high intake (P<0.02) and moderate intake (P<0.05) groups, but this change in birth weight was not associated with corresponding changes in fetal cotyledon mass (76+/-10.3 g). Moreover, the number of fetal cotyledons was similar in all three groups. Thus, progesterone did not directly affect the growth of the fetal cotyledon but may have influenced placental vascularity, blood flow or nutrient transfer capacity or alternatively the development of the embryonic inner cell mass. PMID- 14525531 TI - Intracellular calcium oscillations and activation in horse oocytes injected with stallion sperm extracts or spermatozoa. AB - In oocytes from all mammalian species studied to date, fertilization by a spermatozoon induces intracellular calcium ([Ca(2+)](i)) oscillations that are crucial for appropriate oocyte activation and embryonic development. Such patterns are species-specific and have not yet been elucidated in horses; it is also not known whether equine oocytes respond with transient [Ca(2+)](i) oscillations when fertilized or treated with parthenogenetic agents. Therefore, the aims of this study were: (i) to characterize the activity of equine sperm extracts microinjected into mouse oocytes; (ii) to ascertain in horse oocytes the [Ca(2+)](i)-releasing activity and activating capacity of equine sperm extracts corresponding to the activity present in a single stallion spermatozoon; and (iii) to determine whether equine oocytes respond with [Ca(2+)](i) transients and activation when fertilized using the intracytoplasmic sperm injection (ICSI) procedure. The results of this study indicate that equine sperm extracts are able to induce [Ca(2+)](i) oscillations, activation and embryo development in mouse oocytes. Furthermore, in horse oocytes, injection of sperm extracts induced persistent [Ca(2+)](i) oscillations that lasted for >60 min and initiated oocyte activation. Nevertheless, injection of a single stallion spermatozoon did not consistently initiate [Ca(2+)](i) oscillations in horse oocytes. It is concluded that stallion sperm extracts can efficiently induce [Ca(2+)](i) responses and parthenogenesis in horse oocytes, and can be used to elucidate the signalling mechanism of fertilization in horses. Conversely, the inconsistent [Ca(2+)](i) responses obtained with sperm injection in horse oocytes may explain, at least in part, the low developmental success obtained using ICSI in large animal species. PMID- 14525532 TI - Effects of stage of oestrous cycle and progesterone supplementation during culture on maturation of canine oocytes in vitro. AB - The aim of this study was to evaluate the effect of progesterone supplementation and stage of oestrous cycle on in vitro maturation (IVM) of canine oocytes. Oocytes were cultured in medium supplemented with 0, 2000, 4000 or 8000 ng progesterone ml(-1) (Expt 1; n=274 oocytes) or 0, 20, 200 or 2000 ng progesterone ml(-1) (Expt 2; n=789 oocytes). In Expt 3, oocytes (n=1202) were cultured in a bi phasic system of meiotic arrest followed by IVM, both in the presence of 0, 20, 200 or 2000 ng progesterone ml(-1). Rates of meiotic resumption for Expt 1 ranged from 40.0% to 58.5%; there were no significant differences among groups. In Expt 2, rate of meiotic resumption was significantly lower in the 2000 ng progesterone ml(-1) treatment (35.5%) compared with the 200 ng progesterone ml(-1) treatment (54.0%; P<0.05). There were no significant differences in rates of maturation to metaphase II among treatments in Expt 1 (1.8-8.6%) or Expt 2 (8.4-14.7%); however, oocytes collected from ovaries of bitches in oestrus and dioestrus had higher rates of maturation to metaphase II than did oocytes from bitches at pro oestrus or anoestrus (P<0.01). In Expt 3, no differences were observed in rates of maturation among treatment groups. Rates of maturation to metaphase II of oocytes from bitches in dioestrus were significantly higher than those from bitches in pro-oestrus (P<0.01). These results indicate that supplementation of culture medium with progesterone either during maturation or during meiotic arrest before maturation does not increase the rate of IVM of canine oocytes. However, stage of oestrous cycle is a key factor in the selection criteria for meiotically competent canine oocytes for use in in vitro experiments. PMID- 14525533 TI - Inhibition of in vitro maturation of equine oocytes by interleukin 1 beta via specific IL-1 receptors. AB - Interleukin 1 beta (IL-1 beta) inhibits the LH-induced resumption of meiosis of equine oocytes in vitro. The present study was performed to clarify this inhibitory effect of IL-1 beta by testing increasing concentrations of IL-1 beta, and by measuring the effect of addition of IL-1 receptor antagonist (IL-1RA) to the culture medium. The effect of IL-1 beta on epidermal growth factor (EGF) induced resumption of meiosis was also studied. Cumulus-oocyte complexes (COCs) were collected from subordinate follicles on ovaries obtained from an abattoir. In five distinct experiments, COCs were cultured for 30 h and nuclear maturation of oocytes was evaluated by DNA staining. In Expt 1, seven different media were tested: medium 1 (TCM199+BSA); medium 2 (medium 1+50 ng IL-1 beta ml(-1)); medium 3 (medium 1+eLH); and media 4, 5, 6 and 7 (medium 3 containing 0.1, 1.0, 10.0 and 50.0 ng IL-1 beta ml(-1), respectively). In Expt 2, four different media were tested: medium 1 (TCM199+BSA+eLH); medium 2 (medium 1+50 ng IL-1 beta ml(-1)); and media 3 and 4 (medium 2+IL-1RA at 50 and 100 ng ml(-1), respectively). In Expt 3, three different media were tested: medium 1 (TCM199+BSA+eLH); medium 2 (medium 1+50 ng IL-1RA ml(-1)); and medium 3 (medium 2+50 ng IL-1 beta ml(-1)). In Expt 4, four different media were tested: medium 1 (TCM199+BSA+eLH); and media 2, 3 and 4 (medium 1+IL-1RA at 50, 100 and 150 ng ml(-1), respectively). In Expt 5, three different media were tested: medium 1 (TCM199+BSA+EGF); medium 2 (medium 1+50 ng IL-1 beta ml(-1)); and medium 3 (medium 2+50 ng IL-1RA ml(-1)). In Expt 1, LH alone induced an increase in the rate of in vitro maturation (IVM) of equine oocytes (P<0.05), whereas IL-1 beta alone did not have any effect compared with medium 1. IL-1 beta (50 ng ml(-1)) significantly inhibited the eLH-induced IVM of oocytes (P<0.05) compared with medium 3. A decrease in rate of maturation was observed from a concentration of 10 ng IL-1 beta ml(-1) onwards. In Expt 2, the presence of IL-1RA in the culture medium inhibited the effect of IL-1 beta and restored the rate of oocyte maturation (P<0.05) observed in the presence of LH alone. In Expts 3 and 4 it was demonstrated that IL-1RA alone had no positive effect on the eLH-induced rate of maturation. In Expt 5, IL-1 beta inhibited the EGF-induced resumption of meiosis (P<0.05). The addition of IL-1RA inhibited this effect and restored the rate of oocyte maturation (P<0.05) observed with EGF alone. In conclusion, the present data confirm the inhibitory effect of IL-1 beta on IVM of equine oocytes induced by eLH and demonstrate its inhibitory effect on EGF-induced oocyte maturation. The rate of maturation decreased in a dose dependent way and the lowest rate of maturation was observed at 50 ng IL-1 beta ml(-1) (P<0.05). The use of IL-1RA inhibited these effects, demonstrating that the action of IL-1 beta is receptor-mediated. Moreover, the results clearly show that, in equine species, IL-1 beta is involved in the physiology of COCs by regulating resumption of meiosis. PMID- 14525534 TI - Energy metabolism and intracellular pH in boar spermatozoa. AB - The effect of energy metabolism on intracellular pH was studied in boar spermatozoa using nuclear magnetic resonance (NMR) spectroscopy and confocal microscopy with the pH-sensitive dye seminaphthorhodafluor (SNARF-1). Freshly ejaculated spermatozoa had a high adenylate energy charge (AEC=0.8), which decreased to 0.6 under aerobic conditions and to 0.2 under anaerobic conditions. Correspondingly, no ATP resonances but high AMP resonance were visible in (31)P NMR-spectra of the spermatozoa. When an artificial oxygen buffer (Fluosol) and a purpose-built air supply system were used during (31)P-NMR data acquisition, ATP resonances reappeared whereas the AMP resonance disappeared. Boar spermatozoa kept under aerobic conditions have intracellular compartments that differ markedly in pH, as demonstrated by both (31)P-NMR spectroscopy and confocal microscopy. Using confocal microscopy, the midpiece of the flagellum in which all mitochondria are located was identified as an acidic compartment (pH(i-mp) 6.7). The intracellular pH of both the head (pH(i-h)) and the long principal piece of the flagellum (pH(i-pp)) were 7.2 and, thus, only slightly below the extracellular pH (pH(e) 7.3). Storage of spermatozoa in a glucose-free medium at 15 degrees C when they are immotile slowly shifted the pH(i-mp) from 6.7 to 6.9 within 20 h, whereas pH(i-h) and pH(i-pp) remained unchanged (pH 7.1-7.2). When glucose was present in the medium, all visible compartments of the spermatozoa as well as the medium were acidified to pH 6.2 within 20 h. Under these conditions a resonance at 4.8 mg kg(-1) appeared representing glycerol 3-phosphate. PMID- 14525535 TI - Interaction of bovine granulosa and theca cells in a novel serum-free co-culture system. AB - The objective of this study was to develop a defined culture system in which bovine follicular and granulosa cells are grown in close contact with each other and with the extracellular matrix (ECM) component laminin. Granulosa and theca cells from follicles 4-6 mm in diameter were cultured on either side of laminin coated BioCoat cell culture inserts in a serum-free medium containing 10 ng insulin ml(-1) at plating densities of 10(5) and 3 x 10(5) cells per membrane side. The cells adopted a clumped arrangement, maintained steroidogenic activity for at least 7 days and demonstrated paracrine communication by increased steroidogenesis and enhanced cell survival compared with cells in mono-culture. Co-cultured theca cells secreted significantly more androstenedione compared with cells in mono-culture. Granulosa cell viability was doubled by co-culture with theca cells. Co-cultures at both cell plating densities were responsive to treatment with physiological combinations of either FSH, LH and LR3 insulin-like growth factor I (IGF-I) (treatment A) or FSH, LR3 IGF-I and androstenedione (treatment B). Significantly more androstenedione was secreted in the presence of treatment A compared with controls. In contrast, oestradiol secretion was increased only by treatment B. Progesterone secretion was unaffected by treatment and did not increase during culture. Co-cultures at the higher plating density demonstrated higher theca cell survival and better maintenance of the follicular cell phenotype. In conclusion, this novel co-culture system provides a unique model for the study of paracrine communication between ovarian somatic cells and cell-ECM interactions during follicle growth. PMID- 14525536 TI - Evaluation of the contribution of cyclooxygenase 1 and cyclooxygenase 2 to the production of PGE2 and PGF2 alpha in epithelial cells from bovine endometrium. AB - In ruminants, the production of prostaglandins by the endometrium is critical for recognition of pregnancy. In the absence of an embryonic signal, luteolytic pulses of PGF(2 alpha) are released by the uterus. In contrast, the presence of a viable conceptus reduces the production of PGF(2 alpha) relative to PGE(2) and prevents luteolysis through the release of trophoblastic interferon (IFN-tau). Initially, it was thought that epithelial and stromal endometrial cells were specialized in the production of a single type of prostaglandin. However, purified cell populations of both types of cell can produce PGF(2 alpha) and PGE(2); therefore, selective production of PGF(2 alpha) and PGE(2) must be regulated within each type of cell. Two distinct prostaglandin synthases, cyclooxygenase 1 and cyclooxygenase 2, are involved in prostaglandin production and each may catalyse the production of a different prostaglandin. This possibility was investigated in cultured epithelial cells from bovine endometrium. Cells were treated with oxytocin or arachidonic acid, and expression of cyclooxygenase 1 and cyclooxygenase 2 proteins was monitored over time and correlated with prostaglandin accumulation. Cells were also treated with increasing doses of inhibitors of cyclooxygenase 1 or cyclooxygenase 2 (non steroidal anti-inflammatory drugs; NSAIDs) with or without arachidonic acid or oxytocin: flurbiprofen (0-50 micromol l(-1)) was used as a non-selective inhibitor; valeryl salicylate (0-500 micromol l(-1)) was used as a cyclooxygenase 1 inhibitor and NS-398 (0-1 micromol l(-1)) was used as a cyclooxygenase 2 inhibitor. After stimulation with arachidonic acid or oxytocin, prostaglandin production and expression of cyclooxygenase 2 protein were increased. All inhibitors were able to block basal and stimulated prostaglandin production. These results indicate that in endometrium most, if not all, prostaglandin production is probably processed through cyclooxygenase 2. PMID- 14525537 TI - Disruption of imprinting in cloned mouse fetuses from embryonic stem cells. AB - Cloned mice typically display abnormal development, such as overgrowth of fetuses and placentae. Quantitative expression analysis of eight imprinted genes (H19, Igf2, Igf2r, Air, Peg1/Mest, Peg3, Nuronatin (Nnat) and Ndn) and an alternate transcript of Igf2 (P0) in embryonic stem cloned fetuses and placentae at days 9.5, 12.5 and 17.5 after mating was carried out by real time PCR to investigate whether epigenetic modification of imprinted genes is responsible for overgrowth of the fetus and placental hypertrophy. In addition, the methylation pattern through the bisulphite sequencing method in differentially methylated regions of H19 and Igf2r was examined in day 9.5 fetuses and placentae. The results showed clearly that the expression of H19 gene decreased in cloned fetuses at days 12.5 and 17.5 after mating and in placentae at day 17.5 after mating, and Igf2 was also repressed in fetuses at days 9.5 and 12.5 after mating and in placentae at day 17.5 after mating. In contrast, the transcription of P0, which is a placental specific transcript variant of Igf2, increased at more than four times the control in cloned placenta at day 12.5 after mating. Day 9.5 fetuses that have developed normally revealed only hypermethylated alleles in the H19 differently methylated region (DMR), and both hyper- and hypomethylated alleles in the Igf2r DMR2. These results show that inappropriate reprogramming in some imprinted genes affects the development of cloned embryos, and that aberrant P0 Igf2 transcription in particular may cause the overgrowth of cloned fetuses and placentae. PMID- 14525538 TI - Structure-function analysis of HKE4, a member of the new LIV-1 subfamily of zinc transporters. AB - The KE4 proteins are an emerging group of proteins with little known functional data. In the present study, we report the first characterization of the recombinant human KE4 protein in mammalian cells. The KE4 sequences are included in the subfamily of ZIP (Zrt-, Irt-like Proteins) zinc transporters, which we have termed LZT (LIV-1 subfamily of ZIP zinc Transporters). All these LZT sequences contain similarities to ZIP transporters, including the consensus sequence in transmembrane domain IV, which is essential for zinc transport. However, the new LZT subfamily can be separated from other ZIP transporters by the presence of a highly conserved potential metalloprotease motif (HEXPHEXGD) in transmembrane domain V. Here we report the location of HKE4 on intracellular membranes, including the endoplasmic reticulum, and its ability to increase the intracellular free zinc as measured with the zinc-specific fluorescent dye, Newport Green, in a time-, temperature- and concentration-dependent manner. This is in contrast with the zinc influx ability of another LZT protein, LIV-1, which was due to its plasma membrane location. Therefore we have added to the functionality of LZT proteins by reporting their ability to increase intracellular-free zinc, whether they are located on the plasma membrane or on intracellular membranes. This result, in combination with the crucial role that zinc plays in cell growth, emphasizes the importance of this new LZT subfamily, including the KE4 sequences, in the control of intracellular zinc homoeostasis, aberrations of which can lead to diseases such as cancer, immunological disorders and neurological dysfunction. PMID- 14525539 TI - Functional characterization of alpha-glucan,water dikinase, the starch phosphorylating enzyme. AB - GWD (alpha-glucan,water dikinase) is the enzyme that catalyses the phosphorylation of starch by a dikinase-type reaction in which the beta-phosphate of ATP is transferred to either the C-6 or the C-3 position of the glycosyl residue of amylopectin. GWD shows similarity in both sequence and reaction mechanism to bacterial PPS (pyruvate,water dikinase) and PPDK (pyruvate,phosphate dikinase). Amino acid sequence alignments identified a conserved histidine residue located in the putative phosphohistidine domain of potato GWD. Site directed mutagenesis of this histidine residue resulted in an inactive enzyme and loss of autophosphorylation. Native GWD is a homodimer and shows a strict requirement for the presence of alpha-1,6 branch points in its polyglucan substrate, and exhibits a sharp 20-fold increase in activity when the degree of polymerization is increased from 27.8 to 29.5. In spite of the high variability in the degree of starch phosphorylation, GWD proteins are ubiquitous in plants. The overall reaction mechanism of GWD is similar to that of PPS and PPDK, but the GWD family appears to have arisen after divergence of the plant kingdom. The nucleotide-binding domain of GWD exhibits a closer phylogenetic relationship to prokaryotic PPSs than to PPDKs. PMID- 14525540 TI - The prevalence of reduced zidovudine susceptibility in zidovudine-naive, antiretroviral-experienced HIV-1-infected patients. AB - OBJECTIVES: There is increasing in vitro and in vivo evidence that reduced zidovudine (ZDV) susceptibility is generated by the selective pressure conferred by other nucleoside reverse transcriptase inhibitors (NRTIs). However, the degree to which this occurs in clinical practice remains unclear. We assessed phenotypic and genotypic resistance in ZDV-naive patients with virological failure on stavudine (d4T)-containing regimens, with particular reference to potential cross resistance between d4T and ZDV. METHODS: Patients were identified from a clinical database. Treatment history was confirmed by case note evaluation and discussion with patients. Genotypic and phenotypic analyses were undertaken by Virco (Virco BVBA, Mechelen, Belgium). RESULTS: Sixty-seven drug-experienced, ZDV-naive patients who underwent a resistance test while failing a d4T-containing regimen were identified. Of these patients, 23% had received three or more NRTIs and 42% at least one non-nucleoside reverse transcriptase (RT) inhibitor; 22% had viruses with reduced d4T susceptibility (>1.8-fold resistance), and 25% had viruses with reduced ZDV susceptibility (>4-fold). The most frequently observed RT mutations were identified. A significant correlation was found between susceptibility to d4T and susceptibility to ZDV (r=0.36; P=0.003), and also between virtual resistance to d4T and that to ZDV (r=0.38; P=0.002). CONCLUSIONS: A significant minority of d4T-treated, ZDV-naive patients were found to have viruses with reduced ZDV susceptibility, with a variable association with classical ZDV resistance mutations. These data suggest that cross-resistance between d4T and ZDV may involve novel constellations of mutations. Correlations between d4T and ZDV susceptibilities and resistances further support cross-resistance between NRTIs. PMID- 14525541 TI - Matched case-control study to evaluate risk factors for hyperlactataemia in HIV patients on antiretroviral therapy. AB - BACKGROUND: Lactic acidosis is a life-threatening event during antiretroviral therapy (ART). Hyperlactataemia may be a prelude to acidosis. Our database study suggested that female gender, intercurrent illness and didanosine (ddI)-based regimens may increase risk of lactic acidosis. The aim of this matched case control study was to identify risk factors for hyperlactataemia requiring screening. METHODS: Cases were defined as patients with two consecutive lactate samples > or =3.5 mmol/L taken more than 1 week apart. Cases were matched to two controls on gender, use of ddI and total duration of therapy using a 6-month window on either side. Controls never had raised lactate >2.5 mmol/L. A conditional logistic regression analysis using the PHREG procedure in SAS (SAS Institute Inc, Cary, NC) was performed with a discreet logistic model stratified by matching variables. RESULTS: Twenty-one cases were matched to 42 controls. In the univariate model, current use of stavudine (d4T), total cholesterol >5.3 mmol/L and glucose levels > or =5.2 mmol/L gave increased likelihood of persistent hyperlactataemia. The multivariate model showed current use of d4T to be a significant independent predictor of persistent hyperlactataemia. CONCLUSIONS: The results of this case-control study indicate that, when controlling for ddI use, d4T use is an additional risk factor for hyperlactataemia. PMID- 14525542 TI - Interleukin-7 levels may predict virological response in advanced HIV-1-infected patients receiving lopinavir/ritonavir-based therapy. AB - OBJECTIVES: To examine the relationship between levels of the T-cell regulatory cytokine interleukin-7 (IL-7) and CD4 cell counts during immune reconstitution and to assess its prognostic value in advanced HIV-1-infected patients receiving lopinavir/ritonavir-based therapy. METHODS: Thirty-six HIV-1-infected adults who completed 48 weeks of follow-up visits were included in this prospective study. Patients having failed two or more antiretroviral therapy regimens were treated with lopinavir/ritonavir-based therapy. An enzyme-linked immunosorbent assay was used to determine IL-7 plasma levels, flow cytometry was used to analyse cell surface antigens, and polymerase chain reaction was used to quantify plasma HIV 1. RESULTS: Pretreatment IL-7 levels were elevated in all patients (mean 11.0 pg/mL) and were negatively correlated with CD4 cell counts and age (r=-0.59, P<0.001 and r=-0.57, P<0.001, respectively). During the course of treatment, IL-7 levels decreased by 34% while CD4 cell numbers progressively increased by 88%. Multivariate regression analysis showed that only pretreatment IL-7 levels predicted viral load at 48 weeks when controlling for baseline CD4 cell counts, viral load and patient demographics. CONCLUSIONS: These findings are consistent with regulation of T-cell recovery by IL-7, and suggest that IL-7 measurements might be used to predict virological response. PMID- 14525543 TI - Epidemiological risk factors for hypersensitivity reactions to abacavir. AB - We investigated risk factors for hypersensitivity reactions (HSR) to abacavir in a case-control study. In a multivariate analysis, white race [odds ratio (OR), 5.16; 95% confidence interval (CI), 1.16-22.97] and a higher CD8 cell count at initiation of abacavir (>850 vs. < or =850 cells: OR, 3.74; 95% CI, 1.19-11.77) were found to be significantly associated with the development of HSR. Age, gender, stage of disease, prior antiretroviral exposure and type of concurrent antiretroviral therapy were not associated with HSR. Differences in predisposition to HSR according to ethnicity and baseline CD8 cell count may be explained by the reported MHC genetic associations with HSR. PMID- 14525544 TI - Overcoming subjectivity in assessing facial lipoatrophy: is there a role for three-dimensional laser scans? AB - BACKGROUND: Prevalence and incidence rates of lipodystrophy vary widely and frequently rely upon self- and/or clinician reports. Currently no validated assessment tool for facial lipoatrophy is available. AIMS AND OBJECTIVES: To illustrate that assessment of the severity of facial lipoatrophy by patients and clinicians is subjective. To evaluate the reproducibility of facial three dimensional surface laser scans. METHODS: Twenty-three HIV-positive men were recruited from an inner London HIV outpatient clinic in September 2001. CD4 count, viral load, antiretroviral history and body mass index were recorded. Patients and clinicians independently assessed the severity of facial lipoatrophy on a four-point scale and the level agreement was measured. Seventeen of the 23 patients (73.9%) underwent two scans 1 week apart, which were then superimposed. The volume difference (mm3) and mean difference (mm) between the scans for five regions of the face were measured and compared with the self-reported grade of facial lipoatrophy. RESULTS: For each pair of clinicians (P=0.03, 0.005 and 0.0002, respectively), and for one patient-clinician pair (P=0.004), there was a significant systematic difference between the two sets of gradings of facial lipoatrophy. The level of disagreement was generally higher for patients reporting facial lipoatrophy (n=17) compared to those not reporting it (n=6). The mean volume difference and mean difference between any region were within 200 mm3 and 0.25 mm, respectively. Reproducibility was unaffected by the self-reported grade of facial lipoatrophy. CONCLUSIONS: Assessment of the severity of facial lipoatrophy by patients and clinicians is subjective. Three-dimensional facial laser scans are reproducible and may provide an objective tool for monitoring changes in facial lipoatrophy. PMID- 14525545 TI - Estimating the effect of antiretroviral treatment during HIV seroconversion: impact of confounding in observational data. AB - OBJECTIVE: To assess whether treatment with antiretroviral drugs within the first 3 months of infection with HIV affects medium-term health outcomes. DESIGN AND METHODS: Data from 20 cohorts in Europe and Australia were used Concerted Action on SeroConversion to AIDS and Death in Europe (CASCADE). Analysis was restricted to persons seroconverting in 1988-1998 who started antiretroviral treatment in the first 3 months or 1-2 years from seroconversion. The relationship between times to low CD4 count, AIDS and death and time of initiation of treatment was estimated using proportional hazards models. RESULTS: Seroconversion illness was more common in those who began antiretroviral treatment in the first 3 months (73%) than in those who started treatment within 1-2 years post-seroconversion (33%). Subjects receiving early antiretroviral treatment had times to AIDS and to CD4 counts <200 cells/microL that were intermediate between those of subjects starting treatment within 1-2 years and those of the subset of these subjects starting treatment within 1-2 years who also had a prior CD4 count of >350 cells/microL and no prior AIDS diagnosis. CONCLUSIONS: On the basis of these analyses, the effect of antiretroviral treatment initiation during HIV seroconversion is uncertain. It may result in lower rates of progression compared with starting antiretroviral treatment at 1-2 years, but the early antiretroviral treatment group had a similar or even higher incidence of low CD4 counts and AIDS events than the group who started antiretroviral treatment within 1-2 years with CD4 counts over 350 cells/microL and no prior AIDS diagnosis. Estimates of the effect of early treatment are probably confounded with a number of factors, including, in particular, reasons for treatment initiation. PMID- 14525546 TI - Saquinavir induces stable and functional expression of the multidrug transporter P-glycoprotein in human CD4 T-lymphoblastoid CEMrev cells. AB - BACKGROUND: The multidrug transporter P-glycoprotein (P-gp) is expressed in HIV-1 target cells, in a range of pharmacological barriers and in AIDS-associated tumours. P-gp substrates include HIV-1 protease inhibitors (PIs) and anticancer drugs, which are efficiently effluxed from multidrug-resistant (MDR) cells. OBJECTIVES: The aim of this study was to investigate the effect on human CD4 T lymphoblastoid CEMrev cells of saquinavir and other PIs in terms of P-gp expression and to characterize the functional and biochemical patterns of PI induced P-gp molecules. METHODS: CEMrev cells no longer expressing detectable amounts of P-gp were cultured for a prolonged period in the presence of 10 microg/mL saquinavir (CEMsaq10) and tested for P-gp expression and function. Subsequently, CEMsaq10 cells were transferred into medium containing 15 microg/mL saquinavir (CEMsaq15) and cultured for several months. These cell lines were continuously monitored for P-gp expression, function and immunochemical patterns. A similar strategy was adopted to determine whether other PIs, such as ritonavir and indinavir, were able to induce P-gp expression in CEMrev cells. RESULTS: Compared with the drug-diluent control, the exposure of CEMrev cells to 10 microg/mL saquinavir induced, in a consistent fraction of cells (45-50%), de novo expression of functioning P-gp molecules. The transfer of CEMsaq10 cells to 15 microg/mL saquinavir was associated with a dramatic increase in P-gp expression and function (85-90% of CEMsaq15 cells expressed P-gp and effluxed P-gp dye substrates). These saquinavir-induced P-gp molecules included 75-kDa molecules as well as the classical 170-kDa form of P-gp, suggesting induction of a particular isoform of P-gp termed mini-P-glycoprotein. Conversely, ritonavir and indinavir induced transient P-gp expression in a small percentage of the CEMrev cells. CONCLUSIONS: Treatment of human CD4 T-lymphoblastoid CEMrev cells with saquinavir caused over-expression of functioning P-gp molecules. This de novo acquired MDR phenotype, which differed from that induced by other PIs, was stable, as expression and activity of P-gp were observed in CEMsaq10 and CEMsaq15 cells during prolonged in vitro culturing, even in drug-free conditions. PMID- 14525547 TI - Recurrence of Pneumocystis carinii pneumonia in an HIV-infected patient: apparent selective immune reconstitution after initiation of antiretroviral therapy. AB - Although several studies have reported that it is safe to discontinue secondary Pneumocystis carinii pneumonia (PCP) prophylaxis in patients infected with HIV who experience a sustained immune response as a result of antiretroviral therapy, we describe a patient who developed recurrent PCP <3 months after discontinuing trimethoprim-sulfamethoxazole prophylaxis. He developed disease despite a sustained CD4 T-cell count above 200 cells/microL for more than 3 years while on antiretroviral therapy, as well as an apparent immune reconstitution against disseminated Mycobacterium avium complex (MAC) and Histoplasma capsulatum, for which he also discontinued therapy but without adverse effects. Thus, although increasing evidence continues to indicate that HIV-infected patients receiving combinations of antiretroviral therapies may regain specific immunity against opportunistic infections, our patient's experience suggests that this immune recovery may be selective and incomplete. PMID- 14525548 TI - Response to Clark et al. Assessing limiting factors to the acceptance of antiretroviral therapy n a large cohort of injecting drug users. HIV Med 2003;4:33-37. PMID- 14525552 TI - Testing competing path models linking the biochemical variables in red blood cells from Li+-treated bipolar patients. AB - OBJECTIVES: Red blood cells (RBCs) from Li+-treated bipolar patients have shown abnormalities in intracellular Li+ concentration ([Li+]i), Na+/Li+ exchange rates, and membrane phospholipid levels. Based on Li+-loaded RBC studies, we hypothesized that Li+-treated bipolar patients also have varied intracellular free Mg2+ concentrations ([Mg2+]f) as compared with normotensive patients. We addressed how these experimentally determined values are intercorrelated. Assuming that Li+ treatment alters these biochemical parameters, we provide hypothetical pathways based upon structural equation modeling statistics. METHODS: In RBCs from 30 Li+-treated bipolar patients, we determined [Li+]i, serum [Li+] ([Li+]e), Na+/Li+ exchange parameters, membrane phospholipid levels, [Mg2+]f, and Li+ membrane binding affinities. Comprehensive statistical analyses assessed correlations among the biochemical data. We used path analysis statistics to propose potential pathways in which the data were correlated. RESULTS: We found significant correlations within the three Na+/Li+ exchange parameters and percentage composition of the membrane phospholipids. Additional correlations existed between [Mg2+]f and Vstd, Km, or phospholipid composition, between [Li+]i and percentage of phosphatidylcholine, and between percentage of phosphatidylserine and Km. Based on these findings, we hypothesized and statistically determined the most probable pathway through which these parameters were intercorrelated. CONCLUSIONS: Significant correlations existed between the biochemical parameters that describe the cell membrane abnormality and the Li+/Mg2+ competition hypotheses. Using path analysis statistics, we identified a biochemical pathway by which Li+ may assert its cellular effects. This study serves as an illustrative example how path analysis is a valuable tool in determining the direction of a certain biochemical pathway. PMID- 14525551 TI - An overview of recent findings of the Stanley Foundation Bipolar Network (Part I). AB - AIM AND METHODS: Selected recent findings of the Stanley Foundation Bipolar Network are briefly reviewed and their clinical implications discussed. RESULTS: Daily prospective ratings on the NIMH-LCM indicate a high degree of residual depressive morbidity (three times that of hypomania or mania) despite active psychopharmacological treatment with a variety of modalities including mood stabilizers, antidepressants, and benzodiazepines, as well as antipsychotics as necessary. The rates of switching into brief to full hypomania or mania during the use of antidepressants is described, and new data suggesting the potential utility of continuing antidepressants in the small group of patients showing an initial acute and persistent response is noted. Bipolar patients with a history of major environmental adversities in childhood have a more severe course of illness and an increased incidence of suicide attempts compared with those without. Preliminary open data suggest useful antidepressant effects of the atypical antipsychotic quetiapine, while a double-blind randomized controlled study failed to show efficacy of omega-3 fatty acids (6 g of eicosapentaenoic acid compared with placebo for 4 months) in the treatment of either acute depression or rapid cycling. The high prevalence of overweight and increased incidence of antithyroid antibodies in patients with bipolar illness is highlighted. CONCLUSIONS: Together, these findings suggest a very high degree of comorbidity and treatment resistance in outpatients with bipolar illness treated in academic settings and the need to develop not only new treatment approaches, but also much earlier illness recognition, diagnosis, and intervention in an attempt to reverse or prevent this illness burden. PMID- 14525553 TI - No evidence of attentional deficits in stabilized bipolar youth relative to unipolar and control comparators. AB - OBJECTIVE: The purpose of this study was to determine the presence or absence of attentional problems and prior diagnosis of ADHD in a cohort of stabilized bipolar I relative to unipolar and normal control. METHOD: Indices of attention were obtained from bipolar (n = 44), unipolar (n = 30), and normal controls (n = 45). Measures included: Freedom from Distractibility (FD) Composite Index of the WISC III, Conners' Continuous Performance Test (CPT), Wisconsin Card Sorting Test (WCST), and a checklist measure of subjective cognitive/attentional problems (SIP AV). RESULTS: Bipolar (6.8%), unipolar (10%), and no control youth report a prior diagnosis of ADHD. No significant group or sex differences were observed on FD Composite Index, various CPT indices, or the WCST. Despite normative attentional function by objective testing, subjectively experienced cognitive problems in the clinical probands were reported. CONCLUSIONS: This cohort of well-functioning bipolar youth diagnosed on average 3-4 years prior to assessment do not possess attentional deficits based on a variety of objective tests compared to unipolar or control youth, but self report subjective difficulties in attentional/problem solving ability. In contrast to other authors, we do not find that bipolar youth have high rates of comorbid ADHD. PMID- 14525554 TI - Bipolar disorder. II: personality and age of onset. AB - OBJECTIVES: The aim of this study was to examine whether personality i.e. temperament and character interacts with age of onset in bipolar disorder. METHODS: Bipolar patients were recruited among in- and outpatients from lithium dispensaries of northern Sweden. Patients were diagnosed according to DSM-IV criteria for bipolar disorder type I and II. Temperament and Character Inventory (TCI) was used for measuring personality. TCI was administered to 100 lithium treated bipolar patients and 100 controls. RESULTS: Treatment response was significantly lower (p = 0.005) in patients with early onset compared with late onset. Family history (p = 0.013) and suicide attempts (p = 0.001) were also significantly more common in patients with early onset. Further, patients with early onset were significantly higher (p = 0.045) in the temperament factor harm avoidance (HA) than patients with late onset, but the difference was weak. Patients with early onset had more fear of uncertainty (HA2; p = 0.022) and were more shy (HA3; p = 0.030). Bipolar I patients showed similar results as those in the total bipolar group (I and II), with significantly higher HA (p = 0.019, moderate difference), HA2 (p = 0.015) and HA3 (p = 0.043) in patients with early onset compared with late onset. Bipolar II patients showed no differences between early and late age of onset but the groups are small and the results are therefore uncertain. CONCLUSIONS: Early age of onset in bipolar disorder was correlated to an increase in severity, family history, poorer treatment response and poorer prognosis. Early onset was also correlated to personality. PMID- 14525555 TI - Subsyndromal symptoms assessed in longitudinal, prospective follow-up of a cohort of patients with bipolar disorder. AB - BACKGROUND: Many patients with bipolar disorder (BD) do not regain full function following an acute illness episode, but the extent to which this impairment is the result of persistent symptoms has not been well established. This study examined factors associated with persistent subsyndromal symptoms in a well characterized group of BD patients who were prospectively followed for an average of 3 years. METHODS: Detailed life charting data from 138 patients with BD were reviewed. Patients were categorized into euthymic, subsyndromal or syndromal groups according to the clinical state during their most recent year of follow up. The three groups were then examined with respect to comorbidity, function and treatment received. RESULTS: Patients with subsyndromal symptoms had high rates of comorbid anxiety disorders, and were more likely to have increased rates of eating disorders as well. Patients with subsyndromal symptoms had lower global assessment of function (GAF) scores than euthymic patients, and had as many clinic contacts and medication trials as patients with full episodes of illness. CONCLUSIONS: Persistent subsyndromal symptoms in BD patients are associated with high rates of comorbidity that is important to recognize and treat in order to optimize mood and functioning. PMID- 14525556 TI - Life events and onset of a new phase in bipolar affective disorder. AB - BACKGROUND: There is an increasing focus on the impact of psychosocial factors and stressors on the course of bipolar affective disorder. The life event research has revealed many biases and the results are conflicting. In a prospective study we examined the relationship between life events and affective phases in a group of bipolar patients with a long duration of the disease. METHODS: A group of patients with at least three admissions to hospital for bipolar disorder was followed every 3 months for up to 3 years. At each examination an evaluation of affective phase was made according to the Hamilton Depression Scale, the Newcastle Depression Rating Scale and the Bech-Rafaelsen Mania Rating Scale. Moreover, the patients were rated according to the Paykel Life Events Scale. Their current medical treatment was noted. RESULTS: Fifty-six patients (19 men and 37 women) were included in the study. Women experienced a significantly higher number of life events than men. In 21% of the 353 examinations of women, a new phase was preceded by life events whereas this was the case only in 8% of the 152 examinations of men. In 13% of the male examinations the patients were in a manic phase and in 5% in a depressive phase. In 5% of the female examinations the patients were in a manic phase and in 15% in a depressive phase. Half of the women's depressive phases were preceded by life events, but none of the depressive phases of men. The categories of life events preceding the depressive phases presented a significant overweight of somatic ill health and conflicts in the family. CONCLUSION: We found a gender difference in the course of bipolar affective disorder, as women had a significantly higher number of depressive episodes than men and men had a higher number of manic episodes than women. In bipolar patients with long duration of disease a significant number of depressive episodes in women were preceded by negative life events. Somatic health problems and conflicts in the family were significant factors preceding new depressive phases. PMID- 14525557 TI - Intracerebroventricular administration of ouabain as a model of mania in rats. AB - BACKGROUND: Human bipolar illness is characterized by mood state- and diagnosis associated abnormalities of cellular cation distribution and transport. These include reduced sodium pump activity and expression and increased intracellular sodium. If these alterations are related to the pathophysiology of the disease, rather than secondary or ancillary abnormalities, then one would expect that modeling of these changes in vivo would produce lithium-preventable behavioral abnormalities. METHODS: Ouabain, a potent inhibitor of the sodium pump, was administered intracerebroventricularly to male rats previously fed lithium containing food or plain rat chow. Locomotion was then quantified in an open field. RESULTS: Ouabain increased locomotion 300% over baseline. Lithium pretreatment prevented the ouabain-induced hyperlocomotion response. CONCLUSION: Inhibition of central nervous system sodium pump with ouabain produces a plausible animal model of mania. This model may be useful for preclinical screening of potential mood stabilizers. PMID- 14525558 TI - A manic-depressive symptom self-report in optical scanable format. AB - INTRODUCTION: The Internal State Scale (ISS) is a self-report instrument that allows the simultaneous assessment of both manic and depressive symptoms in individuals with manic-depressive disorder. Prior work indicates that subscales are highly correlated with clinician ratings of mania and depression and provide a discriminant function that identifies individuals in manic/hypomanic, mixed, depressed, and euthymic mood states. A drawback to the ISS is that its items were developed in the visual analogue scale (VAS) format which is labor-intensive to score, particularly with repeat (e.g. daily) administration. A Likert-based format would allow quick and easy optical scanning for which scoring could be automated. METHODS: To compare discriminating properties in Likert versus VAS format we re-analyzed previously collected data and collected new data: (a) VAS based ISS scores from 86 subjects from a prior four-site study were re-analyzed by collapsing scores into 20 and then 10 Likert-based bins to assess loss of precision from collapsing scores, and (b) 24 additional subjects were administered the ISS in VAS and Likert formats to assess loss of precision due to instrument completion factors. RESULTS: Discriminant ability, including kappas and receiver operator characteristic curves, were unchanged across the two formats. Within-subjects reliability was uniformally high across formats. CONCLUSIONS: Likert-based scoring of the ISS can be used without loss of precision, thus making automated scoring of the ISS feasible. This format will be particularly useful for studies that require processing of large numbers of ISSs, such as those that collect frequent ratings over long periods of time and/or those that utilize large samples. PMID- 14525560 TI - Verbal memory in mania: effects of clinical state and task requirements. AB - OBJECTIVES: Manic patients exhibit impaired verbal learning and memory, particularly following longstanding illness. However, it is unclear whether recognition and recall performance are differentially influenced by a manic mood state. METHODS: To examine this issue, we administered the California Verbal Learning Test and symptom-rating scales to inpatients with pure or mixed mania, euthymic outpatients, and healthy comparison subjects. RESULTS: An overall performance difference was identified between groups. Manic and euthymic patients performed more poorly than healthy subjects on recall. However, manic patients performed more poorly than euthymic patients and healthy subjects on recognition. CONCLUSIONS: These results suggest that verbal retrieval deficits are stable vulnerability indicators in bipolar disorder, whereas verbal encoding deficits are manic episode indicators. The known subcortical dysfunction in this disorder may produce stable retrieval deficits while acute mood symptoms attenuate encoding during affective episodes only. PMID- 14525559 TI - Oxcarbazepine: clinical experience with hospitalized psychiatric patients. AB - BACKGROUND: Oxcarbazepine (10-keto-carbamazepine) appears to be better tolerated and simpler to use than carbamazepine. It has antimanic effects but, as its potential clinical usefulness and tolerability in broad samples of psychiatric patients remain to be tested, we reviewed both the pharmacology of oxcarbazepine and our early experience with this new agent among psychiatric inpatients. METHODS: We reviewed medical records of all inpatients given oxcarbazepine in the first 15 months of its use at McLean Hospital. Data analyzed included dosing, presenting illnesses, other medications, clinical changes, and adverse effects. RESULTS: Oxcarbazepine was given to 56 inpatients (1.3% of admissions; 31 women, 25 men) presenting with depression (n = 23), mania (n = 19), or psychosis (n = 14). The discharge daily dose for the 43 patients (76%) taking oxcarbazepine was 831 mg/day, 34% higher in men than women, and fell by 9 mg/year-of-age. Oxcarbazepine was the only putative mood-stabilizing agent given at discharge in 19 of 43 cases (44%). It was discontinued in 20% of patients for apparent inefficacy, and 4% for adverse effects. Changes in CGI and GAF scores were similarly high across illnesses, and unrelated to days of use of oxcarbazepine or its dose. CONCLUSIONS: Oxcarbazepine was well tolerated and simpler to use clinically than its precursor carbamazepine. This agent should be studied in controlled trials to test its efficacy in specific types of major psychiatric disorders, and particularly for long-term maintenance treatment in bipolar disorder. PMID- 14525562 TI - Titanic surgical waiting lists: what lies beneath? PMID- 14525561 TI - Thyrotoxicosis after complete or partial lithium withdrawal in two patients with bipolar affective disorder. AB - OBJECTIVES: To highlight and discuss thyrotoxicosis after lithium withdrawal as a potential complication of lithium therapy for bipolar disorder. CASE REPORTS: Both patients presented developed thyrotoxicosis, the first patient after stopping the lithium completely, and the second patient after a reduction in the lithium dose. CONCLUSIONS: Clinicians should be alert to the possibility of thyrotoxicosis emerging when lithium is being completely or partially withdrawn. Such withdrawal could unmask a latent hyperthyroidism. PMID- 14525563 TI - Laparoscopic staging of upper gastrointestinal malignancy. PMID- 14525564 TI - Sentinel node biopsy in the management of breast cancer. PMID- 14525565 TI - Vocal cord paralysis, synkinesis and vocal fold motion impairment. AB - Felix Semon's 'laws' of vocal cord paralysis were conceived over a century ago, based on the simple concept that abductor function of the recurrent laryngeal nerve was more vulnerable than adductor function. It is now clear that the neuromuscular pathology of laryngeal innervation is much more complex. Whether the nerve has been cut, crushed, stretched, cauterized or otherwise injured, it is seldom completely transected. There might be no detectable vocal cord movement at laryngoscopy, yet, electromyography usually shows at least some activity because of incomplete denervation and/or developing synkinesis. Electrical silence hardly ever persists forever. Disordered vocal fold movement following nerve injury appears to depend on laryngeal synkinesis with disorganized neuromuscular function caused by misdirected regeneration and aberrant reinnervation, sometimes by adjacent nerves. The severity of the injury, abnormal random reinnervation, scar tissue formation and nerve growth-stimulating and inhibiting factors influence the final position of the vocal fold. For a better understanding of neurolaryngological disorders it is no longer sufficient to think merely in terms of 'vocal cord paralysis'. PMID- 14525567 TI - Initial experience of abdominal aortic aneurysm repairs in Borneo. AB - BACKGROUND: Abdominal aortic aneurysms (AAA) repairs are routine operations with low mortality in the developed world. There are few studies on the operative management of AAA in the Asian population.This study reports the initial results from a unit with no previous experience in this surgery by a single surgeon on completion of training. METHODS: All patients with AAA repair from a prospective database between 1996 and 1999 in the south-east Asian state of Sarawak in Borneo Island were analyzed. Three groups were identified on presentation according to clinical urgency of surgery. Elective surgery was offered to all good risk patients with AAA of >or= 5 cm. All symptomatic patients were offered surgery unless contraindicated medically. RESULTS: AAA repairs were performed in 69 patients: 32 (46%)had elective repairs of asymptomatic AAA; 20 (29%) had urgent surgery for symptomatic non-ruptured AAA; and 17 (25%)had surgery for ruptured AAA. The mortality rate for elective surgery was 6%; the two deaths occurred early in the series with the subsequent 25 repairs recorded no further mortality. The mortality rates for the urgent, symptomatic non-ruptured AAA repair and ruptured AAA repair were 20% and 35%, respectively. Cardiac and respiratory complications were the main morbidities.Sixty-three patients seen during this period had no surgery; three presented and died of ruptured AAA, 34 had AAA of or= 5 cm diameter had either no consent for surgery or serious medical contraindications. CONCLUSION: This study showed that AAA can be repaired safely by highly motivated and adequately trained surgeons in a hospital with little previous experience. PMID- 14525566 TI - Is there an association between chronic lung disease and abdominal aortic aneurysm expansion? AB - BACKGROUND: Although there is evidence demonstrating an association between chronic obstructive pulmonary disease (COPD) and abdominal aortic aneurysm (AAA), it is not clear whether COPD predicts greater rates of expansion of established aneurysms. We sought such an association in a cohort of men with aneurysms detected in a population-based study of screening for aneurysms. METHODS: In addition to regular aortic ultrasound scans, 179 men with AAA underwent full lung function testing in order to identify the presence of COPD and its subgroups, emphysema and other obstructive ventilatory defects (OVD). The rate of expansion of each aneurysm was calculated and the men were divided into 'rapid expanders' (3 mm or more per year) and 'slow expanders' (less than 3 mm per year). Any association with the presence of COPD or smoking was tested using a multivariate model. RESULTS: Over a median follow-up period of 36 months the mean rate of aortic expansion for the cohort of 179 men was 2.1 mm/year. There was no significant difference in prevalence of COPD (68% overall) or having ever been a smoker (87% overall) between the rapid expanders and the slow expanders. CONCLUSIONS: Although there was a high prevalence of COPD among men with an AAA, there was no association between the rate of expansion of AAA and the presence of any form of this disease. PMID- 14525568 TI - Effect of transdermal glyceryl trinitrate and anti-inflammatory gel in infusion phlebitis. AB - BACKGROUND: Phlebitis is the commonest complication of intravenous infusion. It has been suggested that it is initiated by venoconstriction at the infusion site, hence treatment with a vasodilator may reduce its incidence. METHODS: A prospective controlled study was carried out on the effect of transdermal glyceryl trinitrate (GTN) and topical anti-inflammatory gel (non-steroidal anti inflammatory drug; NSAID) on the survival of peripheral intravenous infusion in 386 patients. RESULTS: A total of 34.9% (43 out of 123) of the infusions failed in the control group compared with 14.1% (18 out of 127) in the NSAID group (P < 0.05) and 30.8% (43 out of 136) in the GTN group (P < 0.05). In the control group 31 positive cultures were obtained. Twenty-one positive cultures were obtained in the GTN group and four cases of bacterial proliferation were observed in the NSAID group. CONCLUSIONS: Infusion phlebitis is a common problem in hospitalized patients and its incidence can be effectively reduced by NSAI gel and GTN but NSAI gel is more effective than GTN. PMID- 14525569 TI - Role of community diagnostic ultrasound examination in the diagnosis of full thickness rotator cuff tears. AB - BACKGROUND: Diagnostic ultrasound examination has become the most commonly used investigation in Australia for diagnosing rotator cuff tears. The authors felt that the results of such investigations were often inaccurate in their clinical practices. METHODS: The diagnostic ultrasound findings in 336 cases were compared to arthrography, and in 225 cases findings at surgery. RESULTS: A very poor accuracy rate of 0.38 was found for diagnostic ultrasound examination. The sensitivity was 0.24 and specificity was 0.61. CONCLUSIONS: Extreme caution is recommended in the use of diagnostic ultrasound, as currently practised in a general community setting, in diagnosing full-thickness rotator cuff tears. PMID- 14525570 TI - Parathyroid hormone venous sampling prior to reoperation for primary hyperparathyroidism. AB - BACKGROUND: The surgical cure rate for primary hyperparathyroidismis greater than 95%. For those who have recurrent or persistent disease, preoperative localization improves reoperation success rates. Selective parathyroid venous sampling (SPVS) for intact parathyroid hormone is particularly useful when non invasive localization techniques are negative or inconclusive. METHODS: We present all known cases (n = 13)between 1994 and 2002 who had venous sampling for localization a tour institution prior to reoperation for recurrent or persistent primary hyperparathyroidism. Comparison was made with non-invasive localization procedures. Results of invasive and non-invasive localization were correlated with surgical findings. RESULTS: Of the nine reoperated cases, eight had positive correlations between SPVS and operative findings and histopathology. SPVS did not reveal the parathyroid hormone source in one case with negative non-invasive localization procedures. Comparisons between SPVS,computerized tomography (CT), and parathyroid scintigraphy (MIBI)as expressed in terms of true positive (TP), false positive (FP)and false negative (FN) were: SPVS - TP88.8%, FP 0%, FN 11.1%; CT - TP22.2%, FP 22.2%, FN 55.5%; and MIBI - TP33.3%, FP 0%, FN 66.6%. At least seven of the nine operated cases have been cured; another remained normocalcaemic 2 weeks after subtotal parathyroidectomy. CONCLUSION: In our institution SPVS has proven to be a valuable tool in cases with recurrent or persistent primary hyperparathyroidism and negative non-invasive localization procedures. PMID- 14525571 TI - Laparoscopic staging of upper gastrointestinal malignancy. AB - BACKGROUND: Laparoscopic staging (LS) of upper gastrointestinal malignancy has decreased the number of non-curative laparotomies. However, as radiological techniques have improved the value of this invasive staging technique has decreased, with some units either being more selective or abandoning it altogether for certain tumour types. The aim of the present study is to prospectively evaluate the additional utility of LS of upper gastrointestinal malignancy after radiological staging with modern techniques. METHODS: One hundred and six consecutive patients assessed as having potentially curable upper gastrointestinal malignancy after radiological staging underwent LS between April 1999 and June 2001. Laparoscopic findings, outcome at laparotomy and complications were prospectively recorded. RESULTS: Laparoscopic staging detected incurable disease in 28 of the 106 patients (26%). The negative likelihood ratio was 0.36 (95% CI 0.24-0.53). Twenty-seven patients were considered incurable because of findings at laparoscopy and one on the findings of laparoscopic ultrasound. Ten patients underwent open palliative procedures and seven had non therapeutic laparotomies giving a non-curative laparotomy rate of 16%. LS was most useful for primary liver and biliary tract tumours and was least useful for colorectal liver metastases. The most frequent findings denoting incurability were the presence of liver disease (12 cases) and peritoneal metastases (nine cases). Complications occurred in three patients with one death being attributable in part to the laparoscopy. CONCLUSIONS: Laparoscopy was useful in decreasing the number of non-therapeutic laparotomies, but laparoscopic ultrasound gave little additional benefit. The utility of LS was dependent on tumour type and in particular was of marginal benefit for colorectal liver metastases. LS remains a useful staging tool but should be applied selectively. PMID- 14525572 TI - Sentinel node biopsy for breast cancer: using local results for estimation of risk to the patient. AB - BACKGROUND: Sentinel node biopsy (SNB) is being increasingly used but its place outside randomized trials has not yet been established. METHODS: The first 114 sentinel node (SN) biopsies performed for breast cancer at the Princess Alexandra Hospital from March 1999 to June 2001 are presented. In 111 cases axillary dissection was also performed, allowing the accuracy of the technique to be assessed. A standard combination of preoperative lymphoscintigraphy, intraoperative gamma probe and injection of blue dye was used in most cases. Results are discussed in relation to the risk and potential consequences of understaging. RESULTS: Where both probe and dye were used, the SN was identified in 90% of patients. A significant number of patients were treated in two stages and the technique was no less effective in patients who had SNB performed at a second operation after the primary tumour had already been removed. The interval from radioisotope injection to operation was very wide (between 2 and 22 h) and did not affect the outcome. Nodal metastases were present in 42 patients in whom an SN was found, and in 40 of these the SN was positive, giving a false negative rate of 4.8% (2/42), with the overall percentage of patients understaged being 2%. For this particular group as a whole, the increased risk of death due to systemic therapy being withheld as a consequence of understaging (if SNB alone had been employed) is estimated at less than 1/500. The risk for individuals will vary depending on other features of the particular primary tumour. CONCLUSION: For patients who elect to have the axilla staged using SNB alone, the risk and consequences of understaging need to be discussed. These risks can be estimated by allowing for the specific surgeon's false negative rate for the technique, and considering the likelihood of nodal metastases for a given tumour. There appears to be no disadvantage with performing SNB at a second operation after the primary tumour has already been removed. Clearly, for a large number of patients, SNB alone will be safe, but ideally participation in randomized trials should continue to be encouraged. PMID- 14525573 TI - Sentinel node biopsy in breast cancer: validation study and comparison of blue dye alone with triple modality localization. AB - BACKGROUND: Sentinel node biopsy (SNB) for breast cancer patients is a new technique with the potential to provide an accurate staging of the axillary nodal status while avoiding the morbidity of an axillary dissection. The objective of the present study is to validate the use of sentinel node biopsy in a New Zealand hospital and to compare the ability of patent blue dye (PB) alone with triple modality (TM) (preoperative lymphoscintigraphy, intraoperative gamma probe and intraoperative blue dye) to identify the sentinel node. METHODS: A total of 104 patients who had a palpable breast lump that was confirmed to be malignant by radiology and cytology and a clinical diagnosis of stage I or stage II breast cancer, were enrolled for SNB and randomly assigned to triple modality or blue dye alone for the localization of the sentinel node. Axillary dissection was performed after the sentinel node(s) had been removed. RESULTS: There were 63 patients in the PB group and 41 patients in the TM group. Both groups are comparable, with a similar mean age and primary tumour size. A sentinel node was identified in 57 (90%) of the PB group patients and 40 (98%) of the TM group patients. Of these 23 (37%) of the PB group and 23 (56%) in the TM group had axillary nodes positive for malignancy. There was one false negative SNB in the PB group and two false negative results in the TM group. Therefore, the PB group had an accuracy of 98% and a sensitivity of 96% compared to an accuracy of 95% and a sensitivity of 91% for the TM group. CONCLUSION: The results of the present study validate the use of SNB in suitable breast cancer patients. Identification and the accuracy of the sentinel node localization were similar between the two groups. Therefore, in hospital centres without adequate access to a nuclear medical facility, it would be feasible to conduct SNB using blue dye alone. PMID- 14525574 TI - Trauma teams in Australia: a national survey. AB - BACKGROUND: Trauma teams have been associated with improved trauma patient outcomes. The present study seeks to estimate the use of trauma teams in Australian hospitals and describe their medical composition, leadership and criteria for activation. METHODS: Australian public hospitals with more than 100 beds, an emergency department and offering surgical services were identified. A survey assessing the presence, composition and means of activation of a trauma team was mailed to the 'Director, Emergency Department' of all identified hospitals. Three months later, all hospitals were contacted by telephone to complete and verify data collection. RESULTS: Questionnaires were distributed to 130 hospitals. After exclusion of hospitals that did not receive patients with traumatic injuries, and dedicated paediatric tertiary referral centres, 111 hospitals remained for analysis. Of these, 56% had an established trauma team, while 71% of hospitals without a trauma team claimed to have insufficient doctors to form one team. Ninety-five per cent of trauma teams were potentially activated by prehospital paramedic data (field triage). For 92% of trauma teams a combination of anatomical, physiological and mechanistic criteria were required for activation. The most common methods of mobilizing a trauma team were by dispatching a common call onto individual pagers (31%) or by paging trauma team members individually (31%). Fifty-eight per cent of trauma team leaders were emergency medicine specialists/registrars, while 8% of trauma teams were led by surgeons/registrars. Consultant surgeons were members of 23% of trauma teams and 74% of trauma teams consisted of more junior members after hours. Some form of trauma audit was engaged in by 64% of hospitals. CONCLUSIONS: Trauma teams are yet to be utilized by many Australian hospitals that provide trauma care. Australian surgeons presently have limited leadership roles and membership in trauma teams. Trauma audit can be more widely adopted in Australian hospitals. PMID- 14525575 TI - Thromboprophylaxis practice patterns in hip fracture surgery patients: experience in Perth, Western Australia. AB - BACKGROUND: International guidelines recommend that all patients undergoing hip fracture surgery receive specific thromboprophylaxis. The purpose of the present study was to examine current thromboprophylaxis practice patterns in patients undergoing hip fracture surgery at Royal Perth Hospital. METHODS: A total of 129 consecutive patients admitted to Royal Perth Hospital between 4 February and 21 July 2002 for surgical repair of a fractured neck of femur, was studied. The primary outcome was the frequency, type, and duration of thromboprophylaxis use during hospitalization. RESULTS: Mean patient age was 79.4 +/- 13.4 years and 69.8% (90/129) were female. Seventy-four patients (57.8%; 95% confidence interval (CI): 48.8-66.8%) received specific thromboprophylaxis during hospitalization, including 50 patients (39.1%; 95%CI: 30.6-48.1%) who received pharmacological prophylaxis only, three (2.3%; 95%CI: 0.5-6.7%) who received mechanical prophylaxis only, and 21 (16.4%; 95%CI: 10.5-24.0%) who received both mechanical and pharmacological prophylaxis. Of those receiving pharmacological prophylaxis, 35 (49.3%; 95%CI: 37.2-61.4%) received low-molecular-weight heparin, 26 (36.6%; 95%CI: 25.5-48.9%) received low-dose unfractionated heparin, eight (11.3%; 95%CI: 5.0-21.0%) received warfarin, 35 (49.3%; 95%CI: 37.2-61.8%) received aspirin or clopidogrel, and 27 (38.0%; 95% CI: 26.8-50.3%) received combined anticoagulant and antiplatelet prophylaxis. The median duration of mechanical prophylaxis was 8 days (range: 6-12 days) and that of pharmacological prophylaxis was 12 days (range: 6-26 days). When the 32 patients already taking aspirin or warfarin at the time of admission were excluded, only 45 (46.9%; 95%CI: 36.6-57.3%) of the remaining 96 patients received specific thromboprophylaxis. CONCLUSION: Specific thromboprophylaxis remains under-utilized in patients undergoing surgery for hip fracture at Royal Perth Hospital. These data should prompt the implementation of effective strategies to improve thromboprophylaxis practice patterns in high-risk orthopaedic patients. PMID- 14525576 TI - Will MAC be back? AB - BACKGROUND: Microcystic adnexal carcinoma (MAC) was first described as a specific entity in 1982 by Goldstein et al. MAC is a very aggressive tumour, displaying locally invasive tendencies with perineural invasion. For this reason previous studies have suggested that Mohs micrographic surgery is a superior technique in establishing clear margins of excision. However, there is no evidence to suggest that standard local excision is less effective in achieving tumour clearance or a more favourable outcome. METHODS: All patients with MAC were identified from the pathology database. All patients who had undergone local excision were reviewed and included in the study. Histopathology was reviewed by dermatopathologists. RESULTS: We reviewed the outcome of 17 patients treated with local excision. Eleven were female and six were male with a mean age of 70 years, and a mean follow up of 30 months. Two patients in this series experienced recurrence after standard local excision. The mean lateral and depth margins were 7.69 mm (range 0.2-21) and 4.66 mm (range 1-14.5), respectively. Recurrence occurred in two out of three patients who had lateral clearance margins of less than 4 mm. These two patients had evidence of perineural invasion on histopathology. CONCLUSION: In the present study, standard local excision is shown to be effective in tumour clearance for MAC with no recurrences provided there are lateral margins of at least 4 mm. However, a longer period of follow up is required because of the very aggressive locally invasive nature of the tumour. PMID- 14525577 TI - Analysis of the outcomes of a visiting surgical service to small rural communities. AB - BACKGROUND: A team of visiting surgeons has provided regular clinics and day surgery to rural locations in country towns away from resident surgical centres. This format has provided continuity of care for 7 years despite a constantly changing medical workforce. The aim of the present study was to review the results of the group and to compare them against national standards and to provide a model for future outreach programmes. METHODS: All patient diagnoses, procedures and clinical outcomes were recorded prospectively. This record of activity was then collated. District hospital records and clinical notes have been rechecked for complications over a discrete 4 year period. RESULTS: There have been 7419 items of service provided, including 2676 procedures. The diagnostic grouping and subsequent day-surgery activity are consistent with the top 30 surgical separations from all Australian hospitals. These patients have been found suitable to remain in their own home environment for treatment. CONCLUSION: Experienced surgeons operating on selected patients with careful nursing care, in small country hospitals have outcomes similar to urban hospitals. PMID- 14525578 TI - Noise exposure in the orthopaedic operating theatre: a significant health hazard. AB - BACKGROUND: Surgeons working in orthopedic operating theatres are exposed to significant noise pollution due to the use of powered instruments. This may carry a risk of noise-induced hearing loss. The present study was designed to quantify the noise exposure experienced by surgeons and establish whether this breaches occupational health guidelines for workplace noise exposure. METHODS: A sound dosimeter was worn by the operating surgeon during three total hip replacements and two total knee replacements. A timed record of the procedures was kept concurrently. Noise levels experienced during each part of the procedure and total noise exposures were measured. Noise exposures were compared with occupational health guidelines. RESULTS: Noise exposure in total hip replacement averaged 4.5% (1.52-6.45%) of the allowed daily dose (average duration 77.28 min). Total knee replacement exposure was 5.74% (4.09-7.39%) of allowed daily exposure (average duration: 69.76 min). Maximum sound levels approached but did not exceed recommended limits of 110 dBA (108.3 dBA in total hip replacement and 107.6 dBA in total knee replacement). Transient peak sound levels exceeded maximums of 140 dB on multiple occasions during surgery. CONCLUSIONS: Overall total noise dose during orthopaedic surgery was acceptable but orthopaedic surgeons experience brief periods of noise exposure in excess of legislated guidelines. This constitutes a noise hazard and carries a significant, but unquantified risk for noise-induced hearing loss. PMID- 14525579 TI - Clinical categorization for elective surgery in Victoria. AB - AIM: The aim of the present paper was to assess trends in clinician's utilization of urgency categories for elective surgery. METHODS: The present paper reviews the additions to the Victorian elective surgery waiting list for hip replacement and prostatectomy as recorded by the Elective Surgery Information System database. Review of general trends in utilization over two separate 12 month periods were undertaken. RESULTS: There is inconsistency in categorization of patients referred to the waiting list for hip joint replacement and prostatectomy. An increasing trend to categorize patients as semi-urgent (category 2) in preference to non-urgent (category 3) emerged over this period (category creep). Semi-urgent cases might be competing for access within the category 2 band with less urgent cases. CONCLUSIONS: There seems to be an increasing imbalance between demand for and availability of elective surgery for lower urgency elective surgical procedures. This imbalance, characterized by lengthening waiting times, means that not all patients will receive treatment within the clinically recommended waiting times. The variable approach to categorization of urgency suggests that the process lacks objectivity and consensus. Simple clinical tools to assist prioritization are currently being evaluated in Victoria (Australia) and other countries. PMID- 14525580 TI - Parastomal hernia following minimally invasive stoma formation. AB - BACKGROUND: Minimally invasive intestinal stoma formation using a laparoscopic approach or through a trephine, is widely described in published literature. The incidence of parastomal hernia (PH) following a stoma formed without formal laparotomy is not well reported. The present review aims to assess the current data available on minimally invasive stoma formation, with particular reference to the incidence of PH. METHODS: A literature search using the Pubmed Medline database was performed, locating English language articles on minimally invasive stoma formation from 1970 to 2002. The manuscripts were searched manually for further references. RESULTS: The number of published studies describing laparoscopic stoma formation is small (263 stomas) and the follow-up studied were short (none longer than 1 year). The incidence of PH was 0-6.7%. The incidence following a trephine stoma was 6.7-12%, and the number of patients was small (118) and the follow up short (up to 12 months). CONCLUSIONS: The incidence of PH following minimally invasive stoma formation using a trephine or a laparoscopic technique remains unclear. Studies published to date are generally small and the follow up is short. A prospective randomized trial comparing minimally invasive stoma formation with stoma formation with laparotomy, is required. PMID- 14525581 TI - Role of specific amino acids in nutritional support. AB - Attempts have been made over the past decade to evaluate the role of specific amino acids in the nutritional support of surgical patients. Particular attention has been paid to the branched-chain amino acids, glutamine, and a collection of compounds that have been referred to as 'immunonutrients'. The present review provides an overview of the rationale behind the use of these nutrients and reviews information about their role in patients undergoing surgery. In general, although the use of immunonutrients enhances the nutritional status of patients, and although a number of clinical trials have claimed clinical advantages, there are still doubts about their clinical utility and concerns about their high cost. PMID- 14525582 TI - Adrenal surgery: an update. PMID- 14525583 TI - Minimally invasive parathyroidectomy for recurrent or persistent hyperparathyroidism using carbon track localization. AB - BACKGROUND: The present study documents the use of carbon tracking to localize parathyroid adenomas in three patients with persistent or recurrent parathyroid disease. METHODS: Three patients requiring second or third operations for hyperparathyroidism were operated upon after the parathyroid lesion had been localized preoperatively using a suspension of carbon particles in water. The enlarged parathyroid glands were identified by using one or more of the following: computed axial tomography, magnetic resonance imaging, ultrasound or Sestamibi nuclear scan. The lesion was then localized with ultrasound or computed axial tomography and a carbon track was inserted from the lesion to the skin, leaving a small skin tattoo as a marker for the surgeon. Each patient underwent a minimally invasive parathyroid operation. RESULTS: For each of the three patients the recurrent or persistent parathyroid adenoma was successfully identified and removed via a small incision. CONCLUSION: Minimally invasive techniques for primary hyperparathyroidism are changing our approach to parathyroid surgery. However, a minimally invasive technique is less applicable when the disease is persistent or recurrent. Although, when the recurrent or persistent parathyroid adenoma can be identified, localization and carbon tracking have proved useful in allowing the surgeon to remove the lesion via a minimally invasive technique. PMID- 14525584 TI - Control of venous bleeding during dissection near the head of the pancreas. AB - Avulsion injury to portal vein tributaries can result in worrying haemorrhage during pancreatic gastric, and colonic resection. Attempts to control these avulsion injuries can result in extension of the tear into major portal vein tributaries. Pressure on the bleeding point tends to extend the laceration towards the portal vein, which bleeds torrentially when pressure is released for suturing. A technique is described to control the bleeding from these portal vein tributaries. This technique relies on adequate visualization and control of the torn ends of the portal vein tributaries thus enabling the surgeon to precisely suture the disrupted vessel without any collateral damage. This technique has been used on a number of occasions over the last decade with excellent results. PMID- 14525585 TI - Audit and the birth of the abdomino-perineal excision for carcinoma of the rectum. AB - Sir Ernest Miles first performed his radical abdomino-perineal excision for carcinoma of the rectum in 1907. This event was the culmination of almost a decade of audit in which he carefully studied the outcomes of patients with rectal carcinoma. Miles's interpretation of his audit led, in a stepwise manner, to gradual changes in his operative technique and the eventual development of the radical abdomino-perineal operation; the operation that bears his name. The Miles operation and the principles upon which it is based, would lead to an improved outcome for patients with rectal carcinoma. PMID- 14525586 TI - Recurrent giant cell tumour of the maxilla associated with both Paget's disease and primary hyperparathyroidism. PMID- 14525587 TI - Necrotizing fasciitis: an unusual presentation for rectal carcinoma. PMID- 14525588 TI - Small bowel diverticular disease complicated by perforation. PMID- 14525591 TI - Split-liver transplantation: a review. AB - Split-liver transplantation (SLT), a procedure where one cadaver liver is divided to provide for two recipients, offers immediate expansion of the existing cadaver donor pool. To date, the principal beneficiaries of SLT have been adult/pediatric recipient pairs with excellent outcomes reported; however, the current scarcity of cadaver organs has renewed interest in expanding these techniques to include two adult recipients from one adult cadaver donor. Significant obstacles to the widespread application of SLT exist and must be resolved by the transplant community before greater utilization can be realized. This manuscript reviews the historic background, surgical techniques, current results, and obstacles impeding further application of SLT. PMID- 14525592 TI - Preemptive renal transplantation: why not? AB - Dialysis has been the long-established initial choice of treatment for persons with end-stage renal disease. Transplantation before dialysis, or preemptive renal transplantation (PRT), has been controversial because of the paucity of clinical evidence that has clarified the benefits vs. risks of this approach. However, several recent observational analyses indicate that PRT is the optimal strategy to benefit patients requiring renal replacement therapy. This current review will discuss the advantages that are associated with PRT, and will summarize studies that have investigated the impact of the timing of transplantation on outcomes. We will also discuss the utility of PRT from various perspectives, and describe the challenges ahead in expanding the opportunity of PRT for more patients. PMID- 14525593 TI - Prevention of CD40-triggered dendritic cell maturation and induction of T-cell hyporeactivity by targeting of Janus kinase 3. AB - Pharmacological targeting of Janus kinase 3 (JAK3) has been employed successfully to control allograft rejection and graft-vs.-host disease (GVHD). Recent evidence suggests that in addition to its involvement in common-gamma chain (cgamma) signaling of cytokine receptors, JAK3 is also engaged in the CD40 signaling pathway of peripheral blood monocytes. In this study, we assessed the consequences of JAK3 inhibition during CD40-induced maturation of myeloid dendritic cells (DCs), and tested the impact thereof on the induction of T-cell alloreactivity. Dendritic cells triggering through CD40 induced JAK3 activity, the expression of costimulatory molecules, production of IL-12, and potent allogeneic stimulatory capacity. In contrast, JAK3 inhibition with the rationally designed JAK3 inhibitor WHI-P-154 prevented these effects arresting the DCs at an immature level. Interestingly, DCs exposed to the JAK3-inhibitor during CD40 ligation induced a state of hyporeactivity in alloreactive T cells that was reversible upon exogenous IL-2 supplementation to secondary cultures. These results suggest that immunosuppressive therapies targeting the tyrosine kinase JAK3 may also affect the function of myeloid cells. This property of JAK3 inhibitors therefore represents a further level of interference, which together with the well-established suppression of cgamma signaling could be responsible for their clinical efficacy. PMID- 14525594 TI - Effects of combined treatment with CD25- and CD154-specific monoclonal antibodies in non-human primate allotransplantation. AB - The CD154-specific monoclonal antibody (Mab) hu5c8 greatly prolongs allograft survival in primates. The CD25-specific Mab daclizumab has not, to date, been paired with hu5c8. We evaluated the effects of hu5c8 in vitro, alone and in combination with daclizumab on rhesus-mixed lymphocyte reactions (MLRs). We then evaluated therapy with hu5c8 and daclizumab in four monkey renal allograft recipients compared with monkeys untreated or contemporaneously treated with hu5c8 alone. Lymphocyte proliferation in MLR was reduced by both daclizumab and hu5c8, and their combined effects were additive. Rejection-free allograft survival in monkeys treated with both hu5c8 and daclizumab (74-479 days) was not significantly better than animals treated with hu5c8 alone (257-587 days), and one combined therapy animal rejected while still on hu5c8 therapy, a condition not typically seen with hu5c8 monotherapy. Although daclizumab and hu5c8 are additively effective in MLR, they do not appear to be synergistic in vivo in rhesus monkeys. PMID- 14525595 TI - Development of a novel transgenic mouse for the study of interactions between CD4 and CD8 T cells during graft rejection. AB - The goal of this study was the development of a system in which the cooperative interactions between CD4 and CD8 T cells specific for defined peptides from a single minor histocompatibility antigen could be studied. A transgenic mouse strain that expresses chicken ovalbumin (Act-mOVA) on the surface of all cells in the body was produced as a source of tissues containing such an antigen. Skin grafts from Act-mOVA donors were rapidly and completely rejected by wild-type recipients, but only when both CD4 and CD8 T cells were present. CD4 T cells by themselves caused an incomplete form of rejection characterized by rapid but partial contraction of Act-mOVA grafts. CD8 T cells alone caused complete rejection of Act-mOVA skin grafts but only after a long delay. Adoptively transferred ovalbumin-specific TCR-transgenic CD4 and CD8 T cells were stimulated by Act-mOVA graft antigens and CD8 T-cell accumulation in the grafts was enhanced by specific CD4 T cells. These findings, together with the fact that the ligand for ovalbumin peptide-specific CD8 T cells can be detected in Act-mOVA tissues with an MHC-restricted antibody, make this an ideal system for the study of cooperation between CD4 and CD8 T cells. PMID- 14525596 TI - Analysis of intragraft gene and protein expression of the costimulatory molecules, CD80, CD86 and CD154, in orthotopic liver transplant recipients. AB - CD40-CD154 and/or CD28-CD80/86 costimulatory blockade induces long-term allograft survival in numerous animal models. Studies examining the expression of costimulatory molecules during acute cellular rejection (ACR) have been limited to renal and cardiac allografts. The aim of this study was to describe the relationship between intragraft costimulatory molecule expression in OLT recipients and ACR. Forty-five liver biopsies were obtained at reperfusion and day 7. Gene and protein expression of CD80, CD86 and CD154 were analyzed by RT PCR and immunohistochemistry. CD154 protein expression was present in 13 of 18 patients with a RAI score of 4, but in only two of 14 patients with a RAI score of <4. There was a strong association between the RAI score and the presence of CD80 and CD154 immunoreactivity. CD86 protein expression did not correlate with the severity of ACR. In reperfusion biopsies CD154, but not CD80 or CD86, protein expression correlated with the total ischaemic time. There was no association between expression of costimulatory molecule genes and ACR. In conclusion, we have demonstrated an association between CD154 and CD80 protein expression and ACR in orthotopic liver allografts. PMID- 14525597 TI - Ex vivo priming of naive T cells into EBV-specific Th1/Tc1 effector cells by mature autologous DC loaded with apoptotic/necrotic LCL. AB - Posttransplant lymphoproliferative disorders (PTLDs) represent life-threatening complications of bone marrow and solid organ transplantation (SOTx). These are B cell malignancies triggered by Epstein-Barr Virus (EBV) infection in chronically immunosuppressed (IS) recipients. Immunosuppressed EBV seronegative (EBV(-)) organ recipients are at highest risk of developing PTLD owing to the lack of anti EBV memory T cells to control subsequent EBV challenges. Our aim is to establish effective anti-EBV T-cell generation protocols for prevention or treatment of PTLD encountered in SOTx. We have used autologous dendritic cells (DCs) loaded with apoptotic/necrotic lymphoblastoid cell lines (LCLs) to evaluate the ability of such an approach to activate naive T cells in vitro. In EBV(-) individuals, both CD8+ and CD4+ T-cell responses were amplified by this approach, as detected by IFN-gamma ELISPOT and cytotoxicity assays. The CD8+ T cells were poly-specific anti-EBNA3 A, -LMP2 and -BMLF1, with uniform reversion to a CD45RO+/RA-phenotype, decreased CD62L expression, and up-regulation of the activation markers CD28 and CD69. Addition of rhIL-12 improved anti-viral T-cell responses and reduced the functional differences observed between EBV(+) and EBV(-) responders. In conclusion, the DC/LCL method promotes cross-presentation of EBV-associated epitopes and may serve as an effective protocol for the adoptive immunotherapy of PTLD in EBV(-) SOTx patients. PMID- 14525598 TI - Polyomavirus allograft nephropathy: sequential assessment of histologic viral load, tubulitis, and graft function following changes in immunosuppression. AB - Our initial cases of polyoma virus allograft nephropathy (PVAN) received pulse steroids due to anxiety about concomitant acute rejection triggered by the presence of tubulitis. However, our current policy is to reduce immunosuppression in all cases. The aim of this study was to determine whether clinical follow-up in these patient categories shows any differences in: (a) histologic viral load, (b) grade of tubulitis, and (c) graft function. Reduced viral load assessed within 8 weeks was seen in 4/20 (20.0%) biopsies treated initially by increased immunosuppression, compared to 15/19 (83.3%) biopsies treated with reduced immunosuppression (p = 0.001, Fisher's exact test). Yet, >70% reversal of the rise in serum creatinine occurred in only 3/19 (15.8%) and 1/19 (5.3%) patients, respectively, in these two groups. Improved tubulitis was seen in 11/20 (55%) of biopsies treated with steroids, despite the lack of beneficial effect on serum creatinine in 12/19 (63.1%) instances. In biopsies not treated with any change in immunosuppression, the serum creatinine remained stable in 1/5 (20%) and worsened in 4/5 (80%) biopsies. These data demonstrate that in biopsies with PVAN and tubulitis, reduced immunosuppression is more effective in lowering viral load than steroid therapy. Lack of parallelism between viral load, tubulitis grade, and serum creatinine illustrates a complex interplay of viral and alloimmune factors leading to graft injury. PMID- 14525599 TI - BK polyoma virus allograft nephropathy: ultrastructural features from viral cell entry to lysis. AB - BK virions must enter the host cell and target their genome to the nucleus in order to complete their life cycle. The mechanisms by which the virions accomplish these tasks are not known. In this morphological study we found that BK virions localized beneath the host cell cytoplasmic membrane in 60-70-nm, smooth (non-coated) monopinocytotic vesicles similar to, or consistent with, caveolae. In the cytoplasm, the monopinocytotic vesicles carrying virions appeared to fuse with a system of smooth, vesicles and tubules that communicated with the rough endoplasmic reticulum and was continuous with the Golgi system. Membrane-bound single virions and large tubulo-reticular complexes loaded with virions accumulated in paranuclear locations. Occasional nuclei displayed virions within the perinuclear cisterna in association to the perinuclear viral accumulations. Tubular cells with mature productive infection had large nuclei, distended by daughter virions, whereas they lacked significant numbers of cytoplasmic virions. In addition to virally induced cell necrosis, there was extensive tubular cell damage (apoptosis and necrosis) in morphologically non infected tubules. The observed ultrastructural interactions between the BK virions and host cells are remarkably similar to viral cell entry and nuclear targeting described for SV40 virus. PMID- 14525600 TI - Cardiovascular disease reduction in the outpatient kidney transplant clinic. AB - Cardiovascular disease (CVD) is an important cause of death in kidney transplant recipients. Future CVD mortality was estimated by a risk calculator in all patients (n = 439) with a functioning transplant (>6 months), followed at our center. In addition to CURRENT mortality rates, an OPTIMAL rate (adding anti hypertensive and lipid-lowering therapy in uncontrolled patients) and an HISTORIC rate (higher systolic blood pressures and the absence of statin use in our population 5 years ago) were also calculated. Overall, the predicted CURRENT CVD mortality rates are 0.82 (95% CI 0.81-0.83) of HISTORIC rates. Predicted OPTIMAL CVD mortality rates are 0.90 (95% CI 0.87-0.92) of CURRENT rates. To achieve OPTIMAL rates, a 27% increase in blood pressure and lipid-lowering drug use is required. There were few contraindications to these medications, implying that physician prescribing was the major barrier to minimizing risk. Despite OPTIMAL rates, the transplant population's relative risk is 2.3 (median, 95% CI 2.1-2.5) times higher than that in the general population. Therefore, targeted therapy to reduce CVD risk can have substantial benefit, but CVD mortality may continue to exceed that in the general population. PMID- 14525601 TI - Predictors of lung transplant survival in eurotransplant. AB - This study was undertaken to assess the influence of patient/donor and center factors on lung transplantation outcome. Outcomes of all consecutive first cadaveric lung transplants performed at 21 Eurotransplant centers in 1997-99 were analyzed. The risk-adjusted center effect on mortality was estimated. A Cox model was built including donor and recipient age and gender, primary disease, HLA mismatches, patient's residence, cold ischemic time, donor's cause of death, serum creatinine, type of lung transplant, respiratory support status, clinical condition and percentage predicted FEV1. The center effect was calculated (expressed as the standardized difference between the observed and expected survival rates), and empirical and full Bayes methods were applied to evaluate between-center differences. A total of 590 adults underwent lung transplantation. The primary disease (p=0.01), HLA-mismatches (p = 0.02), clinical condition(p < 0.0001) and the patient's respiratory support status (p = 0.05) were significantly associated with survival. After adjusting for case-mix, no between center differences could be found. An in-depth empirical Bayes analysis showed the between-center variation to be zero. Similar results were obtained from the full Bayes analysis. Based on these data, there is no scientific basis to support a hypothesis of possible association between center volume and lung survival rates. PMID- 14525602 TI - Liver transplant recipients older than 60 years have lower survival and higher incidence of malignancy. AB - Older age is not considered a contraindication for liver transplantation, but age related morbidity may be a cause of mortality. Survival and the incidence of the main post-transplant complications were assessed in 111 adult liver transplant recipients. They were divided in two groups according to their age (patients younger than 60 years, n=54; patients older than 60 years, n=57) and both groups were compared. Older patients were more frequently transplanted for hepatitis C (p= 0.03) and hepatocellular carcinoma (p= 0.05) and their liver disease was less advanced (Child-Pugh and MELD scores were significantly lower; p=0.004 and p=0.05, respectively). After transplantation, older patients had a significantly lower survival (p=0.02). Higher age was independently associated with mortality (hazard ratio for each 10-year increase: 2.1; 95% confidence interval: 1.1- 4.0; p=0.02). The incidence of de novo neoplasia and nonskin neoplasia were higher in older patients (p=0.02 and p =0.007, respectively). Malignancy was the cause of death in one patient younger than 60 years and in 12 patients older than 60 years (p =0.002). In multivariate analysis, a higher age and smoking were independently associated with a higher risk of dying of de novo neoplasia. In conclusion, older liver transplant recipients have a significantly lower survival than younger patients. Malignancy is responsible for this decreased survival. PMID- 14525603 TI - De novo non-alcoholic fatty liver disease following orthotopic liver transplantation. AB - Non-alcoholic fatty liver disease (NAFLD) is an increasingly recognized clinico pathologic entity typically associated with obesity, type II diabetes and hyperlipidemia. It has been noted to recur after orthotopic liver transplantation (OLT). We report four patients who developed de novo NAFLD within 3 months of OLT without the typical predisposing factors of diabetes mellitus or obesity. Three of the four patients underwent OLT for hepatitis C-related cirrhosis, and the other for alcoholic cirrhosis. Examination of the liver explants revealed no evidence of steatosis. No surreptitious alcohol use or a drug-induced process could be identified in these patients. Treatment of recurrent hepatitis C infection in one patient with interferon and ribavirin led to sustained suppression of the viral RNA to undetectable levels, but no improvement in histology or liver enzymes. All four patients had histologic evidence of preservation injury on the initial post-OLT biopsies, but the significance of this finding in relationship to the development of NAFLD is unknown. NAFLD can develop without any of the known predisposing conditions after transplantation, and this raises further questions about the pathogenesis of this condition. PMID- 14525604 TI - Psychological distress and adherence to the medical regimen among adolescent renal transplant recipients. AB - This longitudinal pilot study of adolescent renal transplant recipients (a) describes the prevalence of psychological distress, (b) describes the prevalence of nonadherence, and (c) explores the association between the recipient's psychological distress and his/her subsequent medical adherence. Twenty-two adolescents, aged 13-18 years, completed two interviews that were separated by approximately 12 months. Psychological distress was assessed in three domains: symptoms of depression, anxiety, and anger. Adherence was assessed in three domains: medication taking, blood work, and clinic attendance. At the initial interview, 36.4% had symptoms of depression, 36.4% endorsed anxiety, and 18.2% endorsed excessive state anger. Non-adherence rates were 13.6% for medication, 22.7% for blood work, and 50% for missed clinic. At the second interview, nonadherence with medication remained the same and the other domains decreased. Our small pilot sample, however, limited our ability to detect statistically significant changes over time. Predictive analyses demonstrated that adolescents with excessive anger were at greater risk for subsequently missing medications than adolescents without excessive anger. These findings suggest that while symptoms of depression and anxiety are observed among some adolescents with renal transplants, only anger is associated with elevated risk for nonadherence with medication. PMID- 14525605 TI - Anemia: a continuing problem following kidney transplantation. AB - Cardiovascular disease is a leading cause of death among kidney transplant recipients. Anemia, a risk factor for cardiovascular complications among patients with chronic kidney disease, has not been well characterized in kidney transplant recipients. We performed a retrospective cohort study of the prevalence of and factors associated with anemia among 240 patients who underwent kidney transplantation at our institution. The mean hematocrit (Hct) rose from 33% at 1 month after transplantation to 40% at 12 months after transplantation. The proportion of patients with Hct < 36% was 76% at transplantation and 21% and 36%, 1 year and 4 years after transplantation, respectively. Six months after transplantation, women had higher likelihood (OR = 3.61) of Hct < 36%, while higher Hct at 3 months (OR = 0.67 for 1% higher Hct) and diabetes (OR = 0.14) were associated with a lower likelihood of Hct < 36%. Similar associations were seen 12 months after transplantation. Even among patients with Hct < 30%, only 36% had iron studies, 46% received iron supplementation and 40% received recombinant human erythropoietin. Awareness of factors associated with a lower Hct may prompt better anemia screening and management, potentially improving cardiovascular outcomes among kidney transplant recipients. PMID- 14525606 TI - Old-for-old kidney allocation allows successful expansion of the donor and recipient pool. AB - Allocation of kidneys from donors older than 64 years to recipients older than 64 years was started in 1999 to improve use of older donor kidneys. Kidneys are allocated locally without HLA-matching to keep cold ischemia short. We compared survival and rejection rates in elderly patients allocated in the old-for-old program (ESP) to patients aged 60 years and older based on HLA-matching, expected ischemia and waiting time (ETKAS). The 69 ESP patients were older (67.9 +/- 2.5 vs. 63.9 +/- 2.9 years), had older donors (71.2 +/- 3.9 vs. 44.6 +/- 14.5 years) and more HLA-mismatches (4.2 +/- 1.2 vs. 1.6 +/- 1.7) than the 71 ETKAS patients, while ischemia was shorter (7.8 +/- 3.4 vs. 14.2 +/- 5.5 h). ESP and ETKAS had similar graft (1-year: 83.6% vs. 86.9%) and patient survival (85.2% vs. 89.5%). With the introduction of ESP, use of older recipients and donors rose from less than 2% to 16% and 11%, respectively. Incidence of acute rejections was significantly higher in the ESP group (1 year: 43.2% vs. 27.4%) and significantly correlated with the degree of HLA-matching. Introduction of old-for-old allocation allows successful expansion of the donor and recipient pool without affecting patient and graft survival. HLA-matching should not be ignored, as the risk of acute rejection in elderly patients is substantial. PMID- 14525607 TI - Simultaneous pancreas-kidney transplantation utilizing a common arterial conduit: early experience and potential applications. AB - Simultaneous pancreas-kidney transplantation has gained acceptance as a therapeutic modality for patients with end-stage renal disease secondary to diabetes mellitus. In some instances, performing the procedure as conventionally described with renal revascularization from the left iliac vessels and pancreatic arterial inflow from the right iliac vessels may be difficult or undesirable. We describe our experience with an alternate operative technique utilizing a single arterial conduit to vascularize both organs. We believe that this technique may be of use in certain patients undergoing simultaneous pancreas-kidney transplantation. PMID- 14525608 TI - Hepatic venous outflow reconstruction in right lobe living-donor liver graft using recipient's superficial femoral vein. AB - Venous congestion of a liver graft from a life donor is a disastrous complication with a high risk of graft failure. For safety reasons, the middle hepatic vein (MHV) is currently unanimously left with the donor. As this vessel provides major venous draining of the right anterior sector, reconstruction of significant MHV tributaries is controversial. We describe here successful venous outflow reconstruction in adult-to-adult right lobe living-donor liver transplantation (RL-LDLT) using the recipient's superficial femoral vein (SFV). Six months after transplantation, graft function and perfusion are excellent, and the patient is free of venous morbidity related to the harvest of the SFV. PMID- 14525611 TI - Creative publishing or fad: the case of transplant registry analyses. PMID- 14525613 TI - Use of surrogate outcomes for cardiovascular disease after renal transplantation. PMID- 14525614 TI - Rhabdomyolysis associated with laparoscopic live donor nephrectomy and concomitant surgery: a note of caution. PMID- 14525615 TI - Immunoadsorption with tryptophan adsorbers for successful treatment of late steroid-refractory recurrent focal glomerulosclerosis. PMID- 14525616 TI - Bilateral metastasis to the femoral head as the only manifestation of recurrence of gastric adenocarcinoma: a case report. AB - BACKGROUND: Metastases to the bone as the sole manifestation of recurrence of a gastric carcinoma are extremely rare. CASE REPORT: We herein report the case of a 63-year-old man operated a year and a half before for a poorly differentiated gastric carcinoma affecting the fundus, who developed bilateral metastasis to the femoral head as the sole manifestation of recurrence. He was treated with radiotherapy to control pain with a poor response and both femoral heads had to be eventually resected. CONCLUSIONS: We review the literature on the rare occurrence of osseous metastasis from gastric carcinoma and comment briefly on the therapeutic options for these cases. PMID- 14525617 TI - Quality of Life in elderly patients with cancer. AB - The incidence of most types of cancers is age-dependent and the progressive ageing is rapidly increasing the number of elderly people who need treatment for cancer. Elderly patients present peculiar characteristics that make the choice of the correct treatment more difficult and these patients are often undertreated. Moreover, elderly patients are largely underrepresented in cancer treatment trials, and this makes the experimental evidence on this topic even weaker. Health-related Quality of Life (QOL) has been considered as one of the hard end points for clinical cancer research, and treatment of elderly cancer patients represents a typical situation where its assessment can be particularly useful, because the expected toxicity of treatment could be relevant in the discussion of the treatment choice. However, QOL assessment in the elderly is complicated by several unresolved methodological problems (higher frequency of illiteracy, worse compliance with the questionnaires, concomitant diseases, use of instruments not validated in the aged population). Conduct of clinical trials dedicated to elderly patients is now encouraged but there are few published studies. Advanced non-small-cell lung cancer is one of the fields with the largest amount of research on QOL in elderly patients. The ELVIS study demonstrated the efficacy of single-agent chemotherapy, both in terms of QOL and of survival. The MILES study, in which combination chemotherapy was not superior than single agents, showed that baseline QOL is a strong prognostic indicator in these patients. QOL of patients with breast cancer has been another important field in clinical research over the last decades, and interest on this topic in elderly patients is growing, from loco-regional to palliative treatment. In conclusion, some steps have been done in clinical cancer research dedicated to elderly patients, and the role of QOL assessment in this setting is important. However, many methodological problems must be resolved, in order to obtain reliable and useful results. A QOL assessment could also be useful for elderly patients in clinical practice, where it could improve patient-clinician communication: a wider application of properly selected instruments should be recommended. PMID- 14525618 TI - The management of subjective quality of life by short-stay hospital patients: an exploratory study. AB - BACKGROUND: This study tested the homeostatic model of subjective quality of life in a group of 47 short stay patients as they progressed through the stages of hospitalization for surgery. METHOD: Participants completed a questionnaire measuring subjective quality of life, positive and negative affect, self-esteem, optimism and cognitive flexibility, the day prior to admission (T1), two days post-operation (T2) and one week after discharge (T3). Neuroticism and Extroversion were measured at Time 1. RESULTS: All variables remained stable across the three times, apart from positive affect, which dropped significantly post-operation but returned to its previous level post discharge. CONCLUSION: Although the homeostatic model of subjective quality of life was supported at Time 1, the analyses raise doubts about the stability of personality. This finding is consistent with recent discussions of personality. PMID- 14525619 TI - Spatial variation of Anopheles-transmitted Wuchereria bancrofti and Plasmodium falciparum infection densities in Papua New Guinea. AB - The spatial variation of Wuchereria bancrofti and Plasmodium falciparum infection densities was measured in a rural area of Papua New Guinea where they share anopheline vectors. The spatial correlation of W. bancrofti was found to reduce by half over an estimated distance of 1.7 km, much smaller than the 50 km grid used by the World Health Organization rapid mapping method. For P. falciparum, negligible spatial correlation was found. After mass treatment with anti-filarial drugs, there was negligible correlation between the changes in the densities of the two parasites. PMID- 14525620 TI - Cardiovascular diabetology in the core of a novel interleukins classification: the bad, the good and the aloof. AB - BACKGROUND: The impressive correlation between cardiovascular disease and glucose metabolism alterations has raised the likelihood that atherosclerosis and type 2 diabetes may share common antecedents. Inflammation is emerging as a conceivable etiologic mechanism for both. Interleukins are regulatory proteins with ability to accelerate or inhibit inflammatory processes. PRESENTATION OF THE HYPOTHESIS: A novel interleukins classification is described, based on their role in diabetes and atherosclerosis, hypothesizing that each interleukin (IL) acts on both diseases in the same direction--regardless if harmful, favorable or neutral. TESTING THE HYPOTHESIS: The 29 known interleukins were clustered into three groups: noxious (the "bad", 8 members), comprising IL-1, IL-2, IL-6, IL-7, IL-8, IL-15, IL-17 and IL-18; protective (the "good", 5 members), comprising IL-4, IL 10, IL-11, IL-12 and IL-13; and "aloof", comprising IL-5, IL-9, IL-14, IL-16 and IL-19 through IL-29 (15 members). Each group presented converging effects on both diseases. IL-3 was reluctant to clustering. IMPLICATIONS: These observations imply that 1) favorable effects of a given IL on either diabetes or atherosclerosis predicts similar effects on the other; 2) equally, harmful IL effects on one disease can be extrapolated to the other; and 3) absence of influence of a given IL on one of these diseases forecasts lack of effects on the other. These facts further support the unifying etiologic theory of both ailments, emphasizing the importance of a cardiovascular diabetologic approach to interleukins for future research. Pharmacologic targeting of these cytokines might provide an effective means to simultaneously control both atherosclerosis and diabetes. PMID- 14525621 TI - Critical pathways for the management of preeclampsia and severe preeclampsia in institutionalised health care settings. AB - BACKGROUND: Preeclampsia is a complex disease in which several providers should interact continuously and in a coordinated manner to provide proper health care. However, standardizing criteria to treat patients with preeclampsia is problematical and severe flaws have been observed in the management of the disease. This paper describes a set of critical pathways (CPs) designed to provide uniform criteria for clinical decision-making at different levels of care of pregnant patients with preeclampsia or severe preeclampsia. METHODS: Clinicians and researchers from different countries participated in the construction of the CPs. The CPs were developed using the following steps: a) Definition of the conceptual framework; b) Identification of potential users: primary care physicians and maternal and child health nurses in ambulatory settings; ob/gyn and intensive care physicians in secondary and tertiary care levels. c) Structural development. RESULTS: The CPs address the following care processes: 1. Screening for preeclampsia, risk assessment and classification according to the level of risk. 2. Management of preeclampsia at primary care clinics. 3. Evaluation and management of preeclampsia at secondary and tertiary care hospitals: 4. Criteria for clinical decision-making between conservative management and expedited delivery of patients with severe preeclampsia. CONCLUSION: Since preeclampsia continues to be one of the primary causes of maternal deaths and morbidity worldwide, the expected impact of these CPs is the contribution to improving health care quality in both developed and developing countries. The CPs are designed to be applied in a complex health care system, where different physicians and health providers at different levels of care should interact continuously and in a coordinated manner to provide care to all preeclamptic women. Although the CPs were developed using evidence-based criteria, they could require careful evaluation and remodelling according to each system's demands. Additionally, the CPs need to be tested in large-scale, multi level studies in order to thoroughly examine and evaluate their efficacy and effectiveness. PMID- 14525622 TI - The reporting of methods for reducing and detecting bias: an example from the WHO Misoprostol Third Stage of Labour equivalence randomised controlled trial. AB - BACKGROUND: The aim of this article is to explore ways in which selection bias and ascertainment bias can be reduced and investigated in trials, by using the example of a drug trial carried out in both developed and developing countries in hospital delivery wards. METHODS: We describe an innovative and practical design for the boxes for packing the drugs as a way of increasing the security of allocation concealment and blinding. We also assess ascertainment bias using sensitivity analyses, as some unblinding could have occurred due to a potential side effect of one of the drugs. RESULTS: The sensitivity analyses indicated that the conclusions about the relative effects of the treatments could be maintained even in the unlikely worst-case scenarios. CONCLUSIONS: Detailed description of the procedures protecting against common biases and of the assessment of ascertainment bias in this trial should allow readers to confidently appraise and interpret the results obtained. In addition, our experiences will assist others in planning trials in the future. PMID- 14525623 TI - Multifactorial experimental design and the transitivity of ratios with spotted DNA microarrays. AB - BACKGROUND: Multifactorial experimental designs using DNA microarrays are becoming increasingly common, but the extent of the transitivity of cDNA microarray expression measurements across multiple samples has yet to be explored. RESULTS: A strong correlation between direct and transitive inference for significantly differentially expressed genes is demonstrated, using subsets of a dye-swap loop design. CONCLUSIONS: In experimental design, opportunities for transitive inference should be exploited, while always ensuring that comparisons of greatest interest comprise direct hybridizations. PMID- 14525624 TI - Apparent dependence of protein evolutionary rate on number of interactions is linked to biases in protein-protein interactions data sets. AB - BACKGROUND: Several studies have suggested that proteins that interact with more partners evolve more slowly. The strength and validity of this association has been called into question. Here we investigate how biases in high-throughput protein-protein interaction studies could lead to a spurious correlation. RESULTS: We examined the correlation between evolutionary rate and the number of protein-protein interactions for sets of interactions determined by seven different high-throughput methods in Saccharomyces cerevisiae. Some methods have been shown to be biased towards counting more interactions for abundant proteins, a fact that could be important since abundant proteins are known to evolve more slowly. We show that the apparent tendency for interactive proteins to evolve more slowly varies directly with the bias towards counting more interactions for abundant proteins. Interactions studies with no bias show no correlation between evolutionary rate and the number of interactions, and the one study biased towards counting fewer interactions for abundant proteins actually suggests that interactive proteins evolve more rapidly. In all cases, controlling for protein abundance significantly decreases the observed correlation between interactions and evolutionary rate. Finally, we disprove the hypothesis that small data set size accounts for the failure of some interactions studies to show a correlation between evolutionary rate and the number of interactions. CONCLUSIONS: The only correlation supported by a careful analysis of the data is between evolutionary rate and protein abundance. The reported correlation between evolutionary rate and protein-protein interactions cannot be separated from the biases of some protein-protein interactions studies to count more interactions for abundant proteins. PMID- 14525625 TI - Probing the interface in a human co-chaperonin heptamer: residues disrupting oligomeric unfolded state identified. AB - BACKGROUND: The co-chaperonin protein 10 (cpn10) assists cpn60 in the folding of nonnative polypeptides in a wide range of organisms. All known cpn10 molecules are heptamers of seven identical subunits that are linked together by beta-strand interactions at a large and flexible interface. Unfolding of human mitochondrial cpn10 in urea results in an unfolded heptameric state whereas GuHCl additions result in unfolded monomers. To address the role of specific interface residues in the assembly of cpn10 we prepared two point-mutated variants, in each case removing a hydrophobic residue positioned at the subunit-subunit interface. RESULTS: Replacing valine-100 with a glycine (Val100Gly cpn10) results in a wild type-like protein with seven-fold symmetry although the thermodynamic stability is decreased and the unfolding processes in urea and GuHCl both result in unfolded monomers. In sharp contrast, replacing phenylalanine-8 with a glycine (Phe8Gly cpn10) results in a protein that has lost the ability to assemble. Instead, this protein exists mostly as unfolded monomers. CONCLUSIONS: We conclude that valine-100 is a residue important to adopt an oligomeric unfolded state but it does not affect the ability to assemble in the folded state. In contrast, phenylalanine-8 is required for both heptamer assembly and monomer folding and therefore this mutation results in unfolded monomers at physiological conditions. Despite the plasticity and large size of the cpn10 interface, our observations show that isolated interface residues can be crucial for both the retention of a heptameric unfolded structure and for subunit folding. PMID- 14525627 TI - Noninvasive positive-pressure ventilation:the little things do make the difference! PMID- 14525626 TI - Gammadelta T lymphocytes from cystic fibrosis patients and healthy donors are high TNF-alpha and IFN-gamma-producers in response to Pseudomonas aeruginosa. AB - BACKGROUND: Gammadelta T cells have an important immunoregulatory and effector function through cytokine release. They are involved in the responses to Gram negative bacterium and in protection of lung epithelium integrity. On the other hand, they have been implicated in airway inflammation. METHODS: The aim of the present work was to study intracytoplasmic IL-2, IL-4, IFN-gamma and TNF-alpha production by gammadelta and alphabeta T lymphocytes from cystic fibrosis patients and healthy donors in response to Pseudomonas aeruginosa (PA). Flow cytometric detection was performed after peripheral blood mononuclear cells (PBMC) culture with a cytosolic extract from PA and restimulation with phorbol ester plus ionomycine. Proliferative responses, activation markers and receptor usage of gammadelta T cells were also evaluated. RESULTS: The highest production of cytokine was of TNF-alpha and IFN-gamma, gammadelta being better producers than alphabeta. No differences were found between patients and controls. The Vgamma9delta2 subset of gammadelta T cells was preferentially expanded. CD25 and CD45RO expression by the alphabeta T subset and PBMC proliferative response to PA were defective in cystic fibrosis lymphocytes. CONCLUSION: Our results support the hypothesis that gammadelta T lymphocytes play an important role in the immune response to PA and in the chronic inflammatory lung reaction in cystic fibrosis patients. They do not confirm the involvement of a supressed Th1 cytokine response in the pathogenesis of this disease. PMID- 14525628 TI - Acute physiologic effects of nasal and full-face masks during noninvasive positive-pressure ventilation in patients with acute exacerbations of chronic obstructive pulmonary disease. AB - OBJECTIVE: To assess the efficacy of and patient tolerance for nasal and full face masks during noninvasive positive-pressure ventilation (NPPV) with patients suffering acute exacerbations of chronic obstructive pulmonary disease. SETTING: A respiratory medicine ward of a referral hospital. METHODS: Fourteen patients were randomized to 2 groups. Seven used nasal masks and 7 used full-face masks. We used a portable ventilator and recorded arterial blood gases and indices of respiratory muscle effort before and after 15 min of NPPV. Patient tolerance was scored as follows: no tolerance (mask had to be withdrawn before the study period ended) = 0 points; poor tolerance (patient complained of discomfort from the ventilation devices but nevertheless remained compliant) = 1 point; fair tolerance (patient seemed uncomfortable but did not complain) = 2 points; excellent tolerance (patient felt better than before beginning NPPV) = 3 points. RESULTS: The groups were comparable in clinical and pulmonary function variables at baseline. NPPV improved both arterial blood gases and the indices of respiratory effort, with no significant differences between the groups. During NPPV the group that used full-face mask had a greater decrease in respiratory rate, but no other differences. NPPV was well tolerated in both groups. CONCLUSIONS: In patients suffering acute exacerbations of chronic obstructive pulmonary disease NPPV improves arterial blood gases and respiratory effort indices regardless of the type of mask used. PMID- 14525629 TI - Nebulized bronchodilator formulations: unit-dose or multi-dose? AB - BACKGROUND: Nosocomial infections linked to the use of multi-dose bronchodilator nebulizer formulations have been reported in the literature. OBJECTIVE: Survey American hospital respiratory therapy services to determine practice patterns, opinions, and awareness regarding unit-dose and multi-dose bronchodilator formulations. METHODS: A quota sample targeted 4 hospital size categories (0-100 beds, 101-200 beds, 201-400 beds, and > 400 beds) using a listing of general medical/surgical hospitals from the American Hospital Association. Hospitals were contacted via telephone to identify the director of respiratory therapy services, who was invited to complete a 29-item Web-based survey of their hospital practices and their opinions about and knowledge of issues with multi-dose and unit-dose bronchodilator formulations. RESULTS: One thousand forty-seven hospitals were recruited and 409 valid surveys were completed (completion rate 39%). The reported mean +/- SD percentage of unit-dose nebulizer treatments was 80.2 +/- 26.2%. Seventy-two percent (296) of respondents indicated having a policy and procedure manual that deals specifically with nebulized bronchodilator solutions, but only 107 reported having internal monitoring guidelines for compliance with those policies and procedures. Multi-dose bottles of bronchodilator concentrate were used with multiple patients in 77% of cases, and on average 9.7 +/- 8.5 patients were treated with the same multi-dose bottle. Eighty-one percent of respondents reported that treatments from multi-dose bottles are prepared at the bedside. The length of time a multi-dose bottle was kept (after being opened) ranged from 24 hours (8%) to 1 month (11%), and only 3% of respondents reported following manufacturers' recommendations. In the respondents' opinion the chief advantage of multi-dose was cost per dose (84%), and the chief advantage of unit-dose was less risk of contamination (92%). With other factors (therapist time, cost of saline diluent for multi-dose concentrate, dose-error, and contamination) considered, 73% thought that unit-dose vials were more cost-effective. Three hundred thirty-six respondents (82%) thought that a sterile, low-volume (0.5 mL) unit-dose vial of bronchodilator concentrate would be useful, and 249 (74%) of those 336 respondents indicated that such a formulation would replace multi-dose bottles. Only 56% of respondents knew about the evidence regarding the risk of contamination with multi-dose bottles. CONCLUSIONS: Multi-dose bottles of bronchodilator solution are used in approximately 20% of nebulizer treatments, and without strict adherence to infection control procedures they are a potential source of nosocomial infection. A sterile, low-volume unit-dose vial of bronchodilator concentrate would be a useful alternative to multi-dose concentrate for modifying doses or mixing drugs in nebulizer therapy. PMID- 14525630 TI - Effect of intrapulmonary percussive ventilation on mucus clearance in duchenne muscular dystrophy patients: a preliminary report. AB - OBJECTIVE: To determine the effects of intrapulmonary percussive ventilation (IPV) on mucus clearance in tracheostomized Duchenne muscular dystrophy patients. METHODS: We studied 8 patients, 5 of whom had mucus hypersecretion (> 30 mL/d). In a randomized, cross-over study we compared assisted mucus clearance techniques with and without IPV. There were 2 treatment sequences and each patient received 5 consecutive days of each treatment sequence, delivered 3 times a day. One sequence consisted of (1) assisted mucus clearance technique (AMCT, which involves forced expiratory technique and manual assisted cough), (2) endotracheal suctioning, (3) nebulizer administration of 5 mL of 0.9% sodium chloride solution for 5 min, (4) a second AMCT session, (5) endotracheal suctioning, (6) 45 min after the end of the nebulizer treatment a third AMCT session, (7) endotracheal suctioning. The other treatment sequence was the same except that it included IPV during the 5-min nebulizer treatment. The collected secretions were weighed. Vital capacity was measured once, before the treatments. Heart rate, respiratory rate, oxyhemoglobin saturation, end-tidal carbon dioxide, airway resistance, and peak expiratory flow were measured before and at 45 min after the treatments. Mean values were compared using analysis of variance with repeated measures. RESULTS: In patients with hypersecretion the mean +/- SD weight of the collected secretions was significantly higher with IPV (6.53 +/- 4.77 g vs 4.57 +/- 3.50 g, p = 0.01). Heart rate, respiratory rate, oxyhemoglobin saturation, end-tidal carbon dioxide, airway resistance, and peak expiratory flow did not differ statistically between the 2 treatments. CONCLUSIONS: IPV is a safe airway clearance method for tracheostomized Duchenne muscular dystrophy patients, and this preliminary study suggests that IPV increases the effectiveness of assisted mucus clearance techniques. PMID- 14525631 TI - Albuterol aerosol delivered via metered-dose inhaler to intubated pediatric models of 3 ages, with 4 spacer designs. AB - OBJECTIVE: To determine the amount of albuterol, in various particle size ranges, delivered from a hydrofluoroalkane-propelled metered-dose inhaler (Airomir) in 3 models of pediatric intubation (ages 8 months, 4 years, and 16 years) using 4 types of aerosol reservoir: 3 spacers (ACE, AeroChamber HC MV, metal NebuChamber without 1-way valve) and 1 holding chamber (metal NebuChamber with 1-way valve). METHODS: Five reservoirs of each type were tested with albuterol sulfate delivered via metered-dose inhaler that delivers 100 microg of albuterol per actuation. Each reservoir was connected to an endotracheal tube (ETT) that corresponded to the given patient age (8 months = 4 French; 4 years = 5 French; 16 years = 7.5 French) and to a valved system that allowed connection of the ETT to a cascade impactor. Simulated tidal volumes representative of children of the given ages were passed through the reservoir. Both the cascade impactor and the ETT were enclosed within a 100% humidity, 37 degrees C environment. RESULTS: For the total amount of albuterol inhaled onto the impactor, and both the 1.1-4.7 microm and 1.1-3.3 microm inhaled fine-particle fractions, the NebuChamber-with valve showed significantly greater drug delivery than the NebuChamber-without valve, the AeroChamber HC MV, or the ACE (p < 0.001). Among the reservoirs without valves the NebuChamber showed significantly greater delivery than the AeroChamber HC MV or ACE (p < 0.001) for total drug deposition and for both the 1.1-4.7 microm and 1.1-3.3 microm fine-particle fractions. These results were consistent over all age groups. The AeroChamber HC MV had significantly greater delivery (total deposition) than the ACE (p < 0.001), except in the 4-year-old model. There were no significant differences between the AeroChamber HC MV and the ACE for either the 1.1-4.7 microm or the 1.1-3.3 microm fine-particle fraction. CONCLUSION: An aerosol reservoir with 1-way valve positioned between the spacer and the ETT improved the amount of inhaled albuterol 300-900%, compared to the other reservoirs. PMID- 14525632 TI - Buspirone treatment for apneustic breathing in brain stem infarct. AB - We report a case of brainstem infarction resulting in apneustic breathing, which was alleviated with buspirone. We discuss apneusis, review the literature, and speculate about the benefit of serotonin 1A receptor agonists in the treatment of apneusis and other respiratory disorders. PMID- 14525633 TI - Highlights of the Fourth World Congress on Pediatric Intensive Care. PMID- 14525634 TI - Practice of pediatric critical care medicine: results of the Future of Pediatric Education II survey of sections project. AB - OBJECTIVE: To summarize the demographics and practice patterns of the current pediatric critical care workforce and to identify the key workforce issues that may affect the delivery of pediatric critical care services in the future. DESIGN: A questionnaire designed to analyze current pediatric critical care workforce demographics and future workforce trends. SUBJECTS: Pediatric critical care physicians from the United States were identified from the American Academy of Pediatrics Critical Care Section, from a list of physicians certified in pediatric critical care medicine (PCCM) by the American Board of Pediatrics, and from a list of pediatrician members of the Society for Critical Care Medicine. INTERVENTIONS: None. MEASUREMENTS: PCCM physicians were polled regarding board certification, practice characteristics, professional activities, referral patterns, patient profiles, competition, job satisfaction, and projected retirement age. MAIN RESULTS: A total of 805 PCCM physicians completed the survey. When grouped by age, 40% of the responding PCCM physicians were younger than 40 yrs, 49% were 40 to 49 yrs old, and only 11% were 50 yrs of age or older. The younger group had a higher percentage of female pediatricians than the older groups. For all age groups, the largest proportion of time was devoted to direct patient care time in pediatric critical care. This was especially true for the youngest age group that had the largest amount of patient care time devoted to critical care (43%). Time devoted to research was also significantly higher for the younger age group, although very few respondents reported that they have >50% of their time protected for research. For all age groups, those reporting increases in referral volume and referral complexity over the previous 12 months far outnumbered those reporting decreases. The majority of respondents reported being satisfied with their career choice. In general, respondents were more likely to report that too many rather than too few PCCM physicians were currently being trained. Approximately one third of respondents (34%) planned on leaving the field of critical care medicine before retiring from medicine completely. CONCLUSIONS: PCCM physicians were increasingly women and working for >65 hrs/wk, with a good level of job satisfaction. Competition from a variety of sources seems to affect the work of PCCM physicians. The relatively small percentage of time devoted to research, however, is a finding of great concern. PMID- 14525635 TI - Influence of respiratory system impedance on volume and pressure delivered at the Y piece in ventilated infants. AB - OBJECTIVES: Tidal volume (VT) delivered to infants' airways are overestimated and pressure underestimated when measured in the ventilator and not at the Y piece. This study aimed at evaluating the influence of respiratory system impedance on expiratory VT (VTE) and pressure measurement difference. DESIGN: Prospective observational study. SETTING: Pediatric intensive care unit at a university hospital. PATIENTS: Data were collected between February 2000 and October 2001 for 30 infants (range, 1-23 months) ventilated in the pressure-controlled or volume-controlled mode. INTERVENTIONS: Measurements of VTE, pressure obtained at the same time at the Y piece and on the ventilator Servo 300, were collected in ventilated infants. Respiratory system impedance was calculated from data obtained at the Y piece. Circuit compliance was measured in vitro. VTEs were corrected for compressible volume. MEASUREMENTS AND RESULTS: VTEs were overestimated by the Servo 300 in the pressure-controlled and volume-controlled modes (from 5% to 62% of the value displayed on Servo 300). Maximal inspiratory pressures were underestimated by the Servo 300 in the pressure-controlled mode (difference from -2 to +19 cm H(2)O). Measurement difference increased with increasing respiratory system impedance. Ventilator VTE corrected for circuit compliance did not offer a sufficiently accurate estimation of VTE at the Y piece. CONCLUSIONS: VT and pressure measurements must be performed at the Y piece, especially in infants with increased respiratory system impedance (i.e., decreased respiratory system compliance or increased resistance). Correcting VTE for circuit compliance cannot replace measurement of VT at the Y piece. PMID- 14525636 TI - Response to volume resuscitation in children with severe malaria. AB - OBJECTIVES: To examine whether hypovolemia is an important cause of the acidosis in children with severe malaria. DESIGN: Prospective phase 1 study examining the safety of volume expansion using detailed hemodynamic monitoring. SETTING: High dependency unit of Kilifi District Hospital on the coast of Kenya. PATIENTS: Kenyan children admitted with clinical features of severe malaria (impaired consciousness or deep breathing) complicated by acidosis (base excess of less than -8). Three groups were considered: severe malarial anemia plus acidosis if hemoglobin of <5 mg/dL and base excess less than -8; moderate malaria acidosis if the base excess was between -8 and -15; severe malaria acidosis if the base excess was less than -15. INTERVENTIONS: Patients received between 10 and 40 mL/kg of either 0.9% normal saline or 4.5% human albumin solution. MEASUREMENTS AND MAIN RESULTS: A total of 53 children were recruited, and all had evidence of compensated shock at admission, with tachycardia, tachypnea, and prolonged capillary refill time. Mean central venous pressure (se) at admission was 2.9 cm H(2)O (0.5 cm H(2)O); in the severe malaria acidosis group, 44% had hypotension (systolic blood pressure of <80 mm Hg). Improvements of hemodynamic indices and a reduction in acidosis followed administration of either saline or albumin. By 8 hrs, mean central venous pressure had increased to 7.5 cm H(2)O (0.5 cm H(2)O, F = 34.4, p <.001) and was associated with a reduction in mean respiratory rate from 49 to 41 breaths/min (2 to 1 breaths/min, respectively; F = 7.0; p =.009), a reduction in tachycardia from 151 to 141 beats/min (5 to 3 beats/min, respectively; F = 3.4; p =.06), and a reduction in capillary refill time. No child developed evidence of the life threatening complications of pulmonary edema and increased intracranial pressure. CONCLUSIONS: Volume depletion is present at admission in the majority of children with severe malaria complicated by acidosis. Volume expansion corrects the hemodynamic abnormalities and is associated with improved organ function and reduction in acidosis. Formal trials of volume expansion are needed to determine whether volume expansion will reduce mortality. PMID- 14525637 TI - Nosocomial catheter-related bloodstream infections in a pediatric intensive care unit: risk and rates associated with various intravascular technologies. AB - OBJECTIVE: Nosocomial bloodstream infections are associated with increased patient morbidity, mortality, and hospital costs. More than 90% of these infections are related to the use of intravascular catheter devices. This study was done to assess the risk and rates of catheter related-bloodstream infections (CR-BSI) associated with different intravascular technologies in a pediatric intensive care unit population. DESIGN: Retrospective cohort study. SETTING: A 16 bed pediatric intensive care unit in a tertiary children's hospital. STUDY POPULATION: All admissions between July 1997 and December 1999 requiring placement of an intravascular access device for care were examined. Patients with CR-BSI were identified through ongoing surveillance using Centers for Disease Control/National Nosocomial Infections Surveillance System definitions for bloodstream infection. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Of the 2,728 admissions during the review period, 1,043 (38.3%) required placement of an intravascular access device. Bivariate analysis revealed that patients who required intravascular cannulae for extracorporeal life support had a 10-fold increased risk of developing a CR-BSI, and patients requiring vascular access for renal replacement therapy demonstrated a 4-fold increase in the risk of developing CR-BSI compared with the referent group. There was a significant increase in the CR-BSI rate associated with the use of more intravascular access devices per patient admission. Multivariate logistic regression identified the use of extracorporeal life support therapy and the total duration of use of intravascular access devices as significant independent predictors of CR-BSI when controlling for other predictors. CONCLUSION: The use of extracorporeal life support therapy, the presence of multiple intravascular access devices, and the total duration of intravascular access device use were associated with an increase in the rate and risk of developing CR-BSI in our pediatric intensive care unit population. Larger, prospective studies may help elucidate additional factors responsible for these observations. PMID- 14525638 TI - Ventilator-associated pneumonia in intubated children: comparison of different diagnostic methods. AB - OBJECTIVES: To compare different methods for diagnosis of ventilator-associated pneumonia in intubated children. DESIGN: Prospective epidemiologic study. SETTING: Pediatric intensive care unit of a tertiary care university hospital. PATIENTS: All consecutive pediatric intensive care unit patients <18 yrs of age with suspected ventilator-associated pneumonia. INTERVENTIONS: For all patients, the following diagnostic methods were compared: 1) clinical data using Centers for Disease Control criteria; 2) blind protected bronchoalveolar lavage, evaluating quantitative cultures, bacterial index of >5, Gram stain, and presence of intracellular bacteria; and 3) nonquantitative cultures of endotracheal secretions. The reference standard used was clinical judgment of three independent experts (Delphi method) who retrospectively established by consensus the presence of ventilator-associated pneumonia based on clinical, microbiological, and radiologic data. Concordance between each diagnostic method and the reference standard was evaluated by concordance percentage and kappa score. Validity was evaluated using sensitivity, specificity, positive predictive value, negative predictive value, and global value. RESULTS: A total of 30 patients were included in the study. According to the reference standard, ventilator-associated pneumonia occurred in 10 of 30 patients (33%). Bacterial index of >5 in bronchoalveolar secretions showed the best concordance compared with the reference standard (concordance, 83%; kappa, 0.61). Bacterial index of >5 also showed the best validity (sensitivity, 78%; specificity, 86%; positive predictive value, 70%; negative predictive value, 90%; global value, 90%). Intracellular bacteria and Gram stain from bronchoalveolar secretions were very specific (95% and 81%, respectively) but not sensitive (30% and 50%, respectively). Clinical criteria and endotracheal cultures were very sensitive (100% and 90%, respectively) but poorly specific (15% and 40%, respectively). CONCLUSION: Our data show that the most reliable diagnostic method for ventilator associated pneumonia is a bacterial index of >5, using blind protected bronchoalveolar lavage. Further studies should evaluate the validity of all these methods according to the gold standard (autopsy). PMID- 14525639 TI - Recombinant activated factor VII for life-threatening pulmonary hemorrhage after pediatric cardiac surgery. AB - OBJECTIVE: To report intractable life-threatening pulmonary hemorrhage after cardiac surgery in an infant who was treated successfully with recombinant activated factor VII (rFVIIa). DESIGN: Descriptive case report. SETTING: An 18 bed pediatric intensive care unit at a tertiary-care children's hospital. PATIENT: A 10-wk-old child with acute life-threatening pulmonary hemorrhage after cardiac surgery. INTERVENTIONS: General supportive intensive care. MEASUREMENTS AND MAIN RESULTS: Care included mechanical ventilatory support, inotropic support, and concurrent treatment with blood products (packed cells, platelet concentrates, and plasma-derived products), as well as aprotinin and desmopressin to improve hemostasis. The addition of rFVIIa resulted in complete resolution of the hemorrhage. CONCLUSIONS: rFVIIa should be considered as a possible novel therapeutic approach to be used as rescue therapy for patients presenting with massive life-threatening hemorrhage progressing into hemorrhagic shock. Further controlled trials to elucidate the safety of this treatment are warranted. PMID- 14525640 TI - Percutaneous left ventricular "vent" insertion for left heart decompression during extracorporeal membrane oxygenation. AB - OBJECTIVE: Description of a novel method of left ventricular decompression by a percutaneous technique under transthoracic echocardiographic guidance. DESIGN: Case report. SETTING: Supraregional cardiac referral center. PATIENT: PATIENT with end-stage cardiomyopathy. INTERVENTIONS: Percutaneous insertion of a modified Mullins transseptal sheath under transthoracic echocardiographic guidance. MEASUREMENTS AND MAIN RESULTS: Successful decompression of the left ventricle and subsequent orthotopic heart transplantation. CONCLUSIONS: In patients at high risk of bleeding, a percutaneous technique may be useful for left ventricular decompression. PMID- 14525641 TI - Has the time come to rename the Blalock-Taussig shunt? AB - OBJECTIVE: To describe the contribution of Vivien Thomas to the scientific and technical development of the Blalock-Taussig shunt. DESIGN: Review of the literature and of the papers of Vivien Thomas, Alfred Blalock, and Helen Thomas. Literature review was done using MEDLINE, Ovid, and Library of Congress databases for topics including Vivien Thomas, Alfred Blalock, Helen Taussig, and tetralogy of Fallot. Bibliographies of articles were also searched. SETTING: The papers of Thomas, Blalock, and Taussig were reviewed at the Alan M. Chesney Medical Archives at the Johns Hopkins Medical Institutions. PATIENTS/SUBJECTS: None. INTERVENTIONS: None. RESULTS: Vivien Thomas was a primary contributor in both the scientific basis for and the surgical technical aspect of the Blalock-Taussig shunt. He did much of the fundamental studies in Dr. Blalock's laboratory by himself. Blalock was able to do only one practice procedure in the animal model before performing the first surgery in a 15-month-old girl with tetralogy of Fallot. Thomas stood behind Blalock during the procedure to provide advice to Blalock. CONCLUSIONS: Thomas' essential contributions to the development of the Blalock-Taussig shunt remain obscured by the stature of Alfred Blalock and Helen Taussig. This oversight should be corrected. PMID- 14525642 TI - Deep sedation with propofol for children undergoing ambulatory magnetic resonance imaging of the brain: experience from a pediatric intensive care unit. AB - OBJECTIVES: Use of intravenous propofol sedation to facilitate completion of magnetic resonance imaging of the brain in children. DESIGN: Retrospective, cross sectional. SETTING: A university-affiliated pediatric intensive care unit. PATIENTS: A total of 115 children who received intravenous propofol to complete magnetic resonance imaging of the brain January 1 through December 31, 2001. INTERVENTIONS: Intravenous propofol infusion. MEASUREMENTS AND MAIN RESULTS: The mean age was 4.2 +/- 3.1 yrs, and there were 63 boys and 52 girls. Sixty-nine percent of patients belonged to ASA physical status class I, and 31% belonged to ASA class II. All studies were completed with satisfactory image quality. The total dose of propofol used to complete a magnetic resonance image of the brain was 4.3 +/- 1.7 mg/kg body weight. The mean duration of sedation induction was 4.5 +/- 3.5 mins. The mean time to recovery (from the end of the procedure) was 20 +/- 15 mins. The duration of the procedure averaged 39 +/- 20 mins, and the time to discharge from the hospital was 50 +/- 21 mins from the end of the procedure. No episodes of hypoxia, apnea, or a need for artificial airway were noted. Systolic blood pressure decreased 10% +/- 13%, but none of the patients met the criteria for hypotension. A telephone call the next day to the family did not reveal any delayed complications. CONCLUSIONS: Propofol can safely facilitate ambulatory magnetic resonance imaging of the brain in children, and it is associated with brief induction, recovery, and discharge times from the hospital. A drop in blood pressure, although mild and transient, does occur. Therefore, appropriate monitoring and preparedness for cardiorespiratory support are essential. PMID- 14525643 TI - Effect of red blood cell transfusion on oxygen consumption in the anemic pediatric patient. AB - OBJECTIVE: To compare oxygen consumption (Vo(2)) measured by indirect calorimetry before and after a packed red blood cell (PRBC) transfusion in patients with isovolemic anemia. DESIGN: Prospective, repeated-measures clinical study. SETTING: Outpatient pediatric hematology-oncology clinic. PATIENTS: A total of 17 pediatric hematology-oncology outpatients undergoing a PRBC transfusion for a hematocrit of <26%. INTERVENTIONS: Vo(2) was measured by indirect calorimetry before and after a PRBC transfusion. MEASUREMENTS AND MAIN RESULTS: Baseline hematocrit averaged 23% (15.5-25.7%), hemoglobin averaged 8.24 g/dL (5.2 g/dL-9.3 g/dL). Patients received an average of 10.3 mL/kg (2.8-17.5 mL/kg) of PRBC. After PRBC transfusion, all patients had an increase in Vo(2), with a mean increase of 35.09 mL x min(-1) x m(-2) (5-75 mL x min(-1) x m(-2)) or 19% (3.1-52%; p <.001). No significant correlation was found between the pretransfusion hematocrit or the volume of red blood cells administered and the change in Vo(2). No significant change was noted in systolic blood pressure or respiratory rate. There were 14 patients who had a decrease in heart rate after PRBC transfusion, and seven patients who demonstrated an increase in Vo(2) of <10% were compared with patients with a > or =10% change. No significant difference was found in age, height, weight, initial hematocrit, or volume of red blood cells transfused between these two groups. CONCLUSIONS: A significant increase in Vo(2) was noted after a red blood cell transfusion in pediatric patients with isovolemic anemia. These findings suggest that Vo(2) was dependent on the supply of oxygen in this subset of pediatric patients. Responding to increased oxygen delivery by increasing Vo(2) implies that these patients were functioning in a state of relative oxygen deficit and had made physiologic adaptive changes to function in this state. PMID- 14525644 TI - Changes in red blood cell integrity related to infusion pumps: a comparison of three different pump mechanisms. AB - OBJECTIVE: To study the effects of three different infusion pumps on red blood cell (RBC) integrity. SETTING: Laboratory. INTERVENTIONS: Transfusion of packed RBCs using three different pumps (syringe pump, conventional peristaltic pump, and a new volumetric pump with shuttle mechanism). Flow rate was set at 20 mL/hr and duration was 2.5 hrs, simulating a neonatal transfusion. Experiments were repeated in each pump with eight different units of packed RBCs. MEASUREMENTS AND MAIN RESULTS: Plasma hemoglobin, potassium, lactate dehydrogenase, bilirubin, and osmotic fragility were assessed before and after transfusion. There was a significant degree of hemolysis in the RBC bags before transfusion (mean +/- sd for plasma hemoglobin, 5.9 +/- 3.0 g/L; potassium, 40.0 +/- 11.8 mmol/L; lactate dehydrogenase, 64 +/- 38 units/L; total bilirubin,: 1.5 +/- 0.8 microM/L). Overall increase between samples before and after transfusion was 12% for plasma hemoglobin (p =.01, paired Student's t -test), 2% for potassium (p =.03), 20% for lactate dehydrogenase (p =.0001), and 47% for total bilirubin (p =.04). Plasma hemoglobin and lactate dehydrogenase changes were significantly influenced by the type of infusion pump (p =.006 and.01, multiple regression analysis). Regarding these two variables, the new volumetric pump was less hemolytic than the other two pumps. The degree of the transfusion-related RBC changes was also significantly and independently influenced by the storage time of the blood bags with plasma hemoglobin, bilirubin, and osmotic fragility changing less in old bags. CONCLUSIONS: The new pump's shuttle mechanism seems to be less injurious to RBCs compared with a conventional syringe and a peristaltic infusion pump. However, the clinical significance must be related to the hemolytic changes in the RBC units before they are transfused, which are a major source of the overall hemolytic load. PMID- 14525645 TI - Cardiac rescue with enoximone in volume and catecholamine refractory septic shock. AB - In December 2000 and February 2001, two children with suspected meningococcal disease were admitted to our pediatric intensive unit. Their Glasgow Meningococcal Septicaemia Prognostic score was 12 points. General treatment including antibiotics, steroids in case of meningitis, and fluid replacement, was performed. Despite appropriate volume replacement, intubation and ventilation, noradrenaline and adrenaline continuous infusions < or =1.0 microg/kg/min, and additional bolus infusions, cardiac output deteriorated within minutes in both children. Calcium and bicarbonate were given without sustained effect. Echocardiography demonstrated no pericardial effusion and shortening fraction was <10%. External cardiac massage had to be performed immediately in one case for electromechanical uncoupling. Both patients received a bolus of enoximone 2 mg/kg and 5 mg/kg body weight, respectively, followed by a continuous infusion of 20-23 microg/kg/min. Thereafter, both children had an adequate blood pressure and their shortening fraction increased to >30%. Within minutes, the catecholamine infusion could be reduced in both patients. The children completely recovered from their life-threatening situations. In patients with severe prolonged catecholamine and volume refractory endotoxin shock in Waterhouse-Friderichsen syndrome, even with electromechanical uncoupling and complete myocardial arrest, enoximone can immediately restore myocardial contractility. PMID- 14525646 TI - Descending necrotizing mediastinitis in early childhood: favorable outcome after aggressive treatment. AB - OBJECTIVE: To report a case of descending necrotizing mediastinitis (DNM), an unusual illness in childhood that develops as a complication of oropharyngeal infection. DESIGN: Descriptive case report. SETTING: Pediatric intensive care unit. PATIENT: A 19-month-old boy with a DNM that resulted from a pharyngeal abscess. INTERVENTIONS: Aggressive surgical debridement; intravenous therapy with broad-spectrum antibiotics. MEASUREMENTS AND MAIN RESULTS: Simple chest radiograph and computed tomographic scan, and routine culture were used to assess the patient. Administration of broad-spectrum antibiotics and surgical drainage resulted in clinical resolution of symptoms. CONCLUSIONS: An aggressive approach (broad-spectrum antibiotics and surgical debridement) can improve the prognosis of DNM in children. PMID- 14525647 TI - Magnesium sulfate for control of muscle rigidity and spasms and avoidance of mechanical ventilation in pediatric tetanus. AB - OBJECTIVE: To describe the use of intravenous magnesium sulfate for the control of muscle spasms and severe generalized rigidity in a child with moderate to severe tetanus without the need for prolonged deep sedation, mechanical ventilation, or neuromuscular blockade. DESIGN: Case report. SETTING: Pediatric intensive care unit in a tertiary care, university-based children's hospital. INTERVENTIONS: A continuous infusion of magnesium sulfate. MEASUREMENTS AND MAIN RESULTS: We describe a 12-yr-old child with moderate to severe tetanus who was treated with a continuous infusion of magnesium sulfate to control painful muscle spasms and severe generalized rigidity initially refractory to moderate sedation. Muscle spasms and severe generalized rigidity were improved with magnesium sulfate. No adverse effects associated with the use of magnesium sulfate were noted during the monitoring of cardiovascular and respiratory function, reflexes, and serum magnesium concentrations. CONCLUSIONS: An infusion of magnesium sulfate can be utilized to treat muscle spasms and severe generalized rigidity without the need for deep sedation, mechanical ventilation, or neuromuscular blockade. We recommend that magnesium sulfate be considered in the armamentarium of therapeutics utilized to treat muscle spasms and rigidity associated with tetanus, provided the patient's neurologic, cardiovascular, and respiratory status can be closely monitored in the pediatric intensive care unit. PMID- 14525648 TI - Fatal lactic acidosis during antiretroviral therapy. AB - OBJECTIVE: To describe the first pediatric case of fatal lactic acidosis in an antiretroviral-treated child with human immunodeficiency virus (HIV) infection. DESIGN: Case report. SETTING: Pediatric intensive care unit. PATIENTS: A patient with fatal antiretroviral therapy-associated type B lactic acidosis. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We report the case of a 5-yr old girl with HIV infection, receiving ritonavir, stavudine, and didanosine, who presented with a 10-day history of nausea and vomiting. Severe lactic acidosis was found. Her clinical condition worsened, with progressive increase in serum lactate, despite aggressive supportive therapy, including intravenous alkali and continuous arteriovenous hemodiafiltration. CONCLUSIONS: Fatal lactic acidosis is a complication of antiretroviral therapy in pediatric HIV patients, which has not been previously reported in children. Early recognition of mitochondrial dysfunction in these patients could prevent the development of fatal lactic acidosis. PMID- 14525673 TI - Can we stabilize unstable plaque? PMID- 14525674 TI - Effects of cholesterol-lowering foods versus lovastatin on serum lipids and C reactive protein. PMID- 14525675 TI - Diet and C-reactive protein. AB - C-reactive protein (CRP) is an independent predictor of cardiovascular events in healthy individuals and those with pre-existing disease. It also probably contributes to the disease process. CRP levels are higher in obese subjects and this link is almost certainly because of increased insulin resistance. Interventions that alter insulin resistance, such as weight loss, exercise, and conjugated linoleic acid, also alter CRP. Glycemic load is associated with CRP, but there have been no interventions with altered macronutrient composition. In the context of weight loss, macronutrient composition is probably not important. Alcohol lowers CRP, but the mechanism is unknown. The interaction between gender and obesity needs further work, but it appears that obesity has a greater effect on CRP levels in women. PMID- 14525676 TI - Postprandial lipemia and cardiovascular disease. AB - Postprandial lipemia, characterized by a rise in triglyceride-rich lipoproteins after eating, is a dynamic, nonsteady-state condition in which humans spend the majority of time. There are several lines of evidence suggesting that postprandial lipemia increases risk of atherogenesis. Clinical data show a correlation between postprandial lipoproteins and the presence/progression of coronary artery disease and carotid intimal thickness. Mechanistic studies demonstrate that triglyceride-rich lipoprotein remnants may have adverse effects on endothelium and can penetrate into the subendothelial space. Exchange of core lipids between postprandial lipoproteins and low-density lipoprotein (LDL)/high density lipoprotein (HDL) is increased during prolonged lipemia, resulting in small, dense LDL particles and reduced HDL cholesterol levels. Hemostatic variables, including clotting factors, platelet reactivity, and monocyte cytokine expression, may be increased during postprandial lipemia. Collectively, these data suggest that assessment and treatment of atherosclerosis should include parameters related to postprandial lipemia. PMID- 14525677 TI - Dietary fat and postprandial lipids. AB - Impaired clearance of chylomicron remnants is associated with increased risk of atherosclerosis and cardiovascular disease. An intake of 40 to 50 g of fat in a meal results in significant lipemia in healthy adults, with consecutive fat containing meals enhancing the lipemia. This would suggest that limiting fat intake to approximately 30 g on each eating occasion would minimize postprandial lipemia. Sedentary behavior and obesity independently impair the postprandial metabolism of lipids. Postprandial lipemia causes endothelial dysfunction and results in a transient increase in factor VII activated (FVIIa) concentration. Plasminogen activator inhibitor type-1 activity is associated with fasting plasma triacylglycerol concentration, but is not influenced by postprandial lipemia. Trans-18:1 acid appears to increase cholesterol ester transfer activity acutely compared with oleate. Randomized stearic acid-rich fats result in less postprandial lipemia and a lower postprandial increase in FVIIa, whereas unrandomized cocoa butter results in similar postprandial lipemia and increases in FVIIa compared with oleate. A background diet containing in excess of 3 g/d of long-chain omega-3 fatty acids decreases postprandial lipemia by stimulating lipoprotein lipase expression and decreasing very low-density lipoprotein synthesis, but a diet enriched in alpha-linolenic acid (up to 9.5 g/d) does not show these effects. Future research on diet and postprandial lipids needs to exploit newly gained knowledge on the regulation of adipocyte metabolism by adipokines and nuclear hormone receptors, particularly with regard to fat patterning and reverse cholesterol transport. PMID- 14525678 TI - The postprandial effects of dietary antioxidants in humans. AB - Traditional risk factors as measured in the fasted individual are reported to be responsible for the prediction of only half of the incident cases of cardiovascular disease. However, many complex and deleterious reactions occur in the postprandial state. The consequences of oxidative reactions occurring during this time represent major risk for fatal and nonfatal heart disease, ischemia, and stroke, and include oxidative modifications to low-density lipoproteins (LDL), decreased production and bioactivity of nitric oxide (NO) in endothelial cells, and reduced endothelial function. Supplementation with antioxidants may prevent or reduce many of these risks. Antioxidants have been shown to reduce oxidative modification to LDL cholesterol, prevent glucose auto-oxidation, improve the bioactivity of NO, and attenuate or prevent the decrease in endothelial function associated with the postprandial state. Because many nonfasting reactions represent major risk for disease, postprandial risk analysis must form a larger part of the diagnostic strategy for disease prevention in the future. PMID- 14525679 TI - The concept of apolipoprotein-defined lipoprotein families and its clinical significance. AB - Classification of plasma lipoproteins on the basis of apolipoprotein (apo) composition recognizes two lipoprotein (Lp) classes, one of which is characterized by apoA-I and the other by apoB as major protein constituents. The former lipoprotein class consists of three major subclasses referred to (according to their apolipoprotein constituents) as Lp-A-I, Lp-A-I:A-II, and Lp-A II, and the latter one of five subclasses called Lp-B, Lp-B:E, Lp-B:C, Lp-B:C:E, and Lp-A-II:B:C:D:E. As polydisperse systems of particles, the apoA-I-containing lipoproteins overlap in high-density segments and apoB- containing lipoproteins in low-density segments of the density gradient. Each subclass is characterized by a specific chemical composition and metabolic property. Normolipidemia and dyslipoproteinemias are characterized by quantitative rather than qualitative differences in the levels of apoA- and apoB-containing subclasses. Furthermore, apoA-containing subclasses seem to differ with respect to their relative antiatherogenic capacities, and apoB-containing subclasses regarding their relative atherogenic potentials. Whereas Lp-A-I may have a greater antiatherogenic capacity than other apoA-containing subclasses, the cholesterol enriched Lp-B:C appears to be the most atherogenic subclass among apoB-containing lipoprotein families. The use of pharmacologic and/or dietary interventions to treat dyslipoproteinemias has already shown that these therapeutic modalities may affect selectively individual apolipoprotein-defined lipoproteins, and thus allow the selection of individualized treatments targeted at decreasing harmful and/or increasing beneficial lipoprotein subclasses. PMID- 14525680 TI - Soy, garlic, and ginkgo biloba: their potential role in cardiovascular disease prevention and treatment. AB - There are hundreds of foods, nutrients, herbs, and botanicals that have "bioactive" constituents with potential human health benefits. Three of these are discussed in this review: soy, garlic, and ginkgo biloba. Each of these three choices involves an ingestible item composed of a complex mixture of bioactive agents. For each of these three, there is a large and growing body of research suggesting potential cardiovascular health benefits. And for each there is at least some level of disagreement or controversy. The focus of this review is on results from recent human clinical trials. PMID- 14525681 TI - A review of low-carbohydrate ketogenic diets. AB - In response to the emerging epidemic of obesity in the United States, a renewal of interest in alternative diets has occurred, especially in diets that limit carbohydrate intake. Recent research has demonstrated that low-carbohydrate ketogenic diets can lead to weight loss and favorable changes in serum triglycerides and high-density lipoprotein cholesterol. This review summarizes the physiology and recent clinical studies regarding this type of diet. PMID- 14525682 TI - The DASH diet and blood pressure. AB - High blood pressure (also called hypertension) is one of the most important and common risk factors for atherosclerotic cardiovascular disease (CVD) and other chronic diseases. National guidelines recommend that all individuals with blood pressure readings of 120/80 mm Hg or higher adopt healthy lifestyle habits, including the Dietary Approaches to Stop Hypertension (DASH) diet, to manage their blood pressure. The DASH diet, which is high in fruits, vegetables, and low fat dairy products and reduced in fat, has been shown in large, randomized, controlled trials to reduce blood pressure significantly. The DASH diet also has been shown to reduce blood cholesterol and homocysteine levels and to enhance the benefits of antihypertensive drug therapy. The DASH diet should be promoted, along with maintaining healthy weight, reducing sodium intake, increasing regular physical activity, and limiting alcohol intake, for lowering blood pressure and reducing the risk of CVD. PMID- 14525683 TI - Dietary intake of fruits and vegetables and risk of cardiovascular disease. AB - Cardiovascular disease (CVD) remains the leading cause of morbidity and mortality in the United States and worldwide. In this review, we examine the scientific evidence in support of current dietary recommendations to increase fruit and vegetable intake for CVD prevention. Available evidence indicates that persons who consume more fruits and vegetables often have lower prevalence of important risk factors for CVD, including hypertension, obesity, and type 2 diabetes mellitus. Recent large, prospective studies also show a direct inverse association between fruit and vegetable intake and the development of CVD incidents such as coronary heart disease and stroke. However, the biologic mechanisms whereby fruits and vegetables may exert their effects are not entirely clear and are likely to be multiple. Many nutrients and phytochemicals in fruits and vegetables, including fiber, potassium, and folate, could be independently or jointly responsible for the apparent reduction in CVD risk. Functional aspects of fruits and vegetables, such as their low dietary glycemic load and energy density, may also play a significant role. Although it is important to continue our quest for mechanistic insights, given the great potential for benefits already known, greater efforts and resources are needed to support dietary changes that encourage increased fruit and vegetable intake. PMID- 14525684 TI - Dietary fiber and coronary disease: does the evidence support an association? AB - Large, prospective, epidemiologic studies show a protective effect of dietary fiber against coronary heart disease (CHD) and form the basis for new recommendations from the National Academy of Science for fiber intake (38 and 25 g/d for young men and women, respectively, based on an intake of 14 g of fiber per 1000 kcal). Mechanisms by which fibers may protect against CHD include lowering blood cholesterol (soluble fibers), attenuating blood triglyceride levels (mostly soluble fibers), decreasing hypertension (all fibers), and normalizing postprandial blood glucose levels (all fibers). An important consideration in making diet recommendations to protect against CHD is that the total amount of fiber from fiber-containing foods is important, and individuals should not just be counseled to focus on soluble fiber. PMID- 14525686 TI - [Advantages and limitations of experimental animal models for the study of infectious diseases]. PMID- 14525685 TI - Assessing endothelial function as a risk factor for cardiovascular disease. AB - The vascular endothelium plays a key role in the prevention of atherosclerosis. Endothelial dysfunction is an antecedent of clinical cardiovascular disease and can be viewed as the final pathway between coronary risk factors and the development of atherosclerosis. The development of a noninvasive method of assessing endothelial health (ie, measurement of flow- mediated dilation of the brachial artery) has enabled investigators to evaluate the effects of dietary patterns on vascular function. Emerging evidence indicates that dietary fats may acutely impair endothelial function, but this effect is dependent on the type of fat and on the other nutrients consumed. Although inconclusive, studies in cohorts of modest size suggest that antioxidants, L-arginine, and folic acid may modulate endothelial function. Additional research is needed to define the impact of complex, long-term dietary patterns on the vascular endothelium. PMID- 14525687 TI - [Experimental meningitis due to a high-level cephalosporin-resistant strain of Streptococcus pneumoniae serotype 23F]. AB - INTRODUCTION: The increasing prevalence of high-level cephalosporin-resistant strains of Streptococcus pneumoniae could complicate the treatment of severe infections such as meningitis. There are still questions as to the characteristics of these strains, their ability to produce severe infection, and the inflammatory response they induce in CSF. METHODS: Using a rabbit model of meningitis, we sought to determine the pathogenicity and differences in inflammatory parameters in two serotype 23F S. pneumoniae strains with different susceptibility to betalactams. Minimal inhibitory concentrations of the two strains were as follows: strain A--PEN 4 micro g/mL, CRO/CTX 2 micro g/mL--and strain B--PEN 0.12 micro g/mL, CRO/CTX 32 micro g/mL. RESULTS: Strain A resulted in a greater incidence of secondary bacteremia and higher inflammatory parameters during the early phases of infection. Strain B caused brain edema, a more severe inflammatory response and significantly higher mortality at the end of the experiment. CONCLUSIONS: Both strains induced meningitis in the animal model. The differences in inflammatory response produced by the two strains could be related to the variations that determine the betalactam resistance level. PMID- 14525688 TI - [Bacteremia due to Pasteurella spp.: a rare process in our hospital over the last 8 years]. AB - OBJECTIVES: To review and update the epidemic and clinical knowledge concerning disseminated blood disease caused by Pasteurella species in our area. METHODS: Retrospective study of Pasteurella species bacteremia (PSB) episodes occurring in patients attended from January 1994 to December 2001 in a single tertiary hospital. RESULTS: Among the 31 clinical samples remitted to the Microbiology Laboratory in which a species of Pasteurella was identified, 5 (16%) corresponded to positive blood cultures in 5 patients. Pasteurella multocida was the predominant species, identified in 70% of all isolations and all but one positive blood culture. All the patients were adults over 50 years old and all had underlying illnesses causing comorbidity or some degree of immunocompromise, with cardiovascular and hypertensive conditions being the most frequent; only one patient had liver cirrhosis. In all cases, except one, contact or coexistence with dogs or cats was documented. The clinical presentation of PSB was non specific and only two episodes were related with a possible focal, soft-tissue origin. There were no serious complications, such as septic shock, organ failure or invasive disease (meningitis or endocarditis). All patients cured with antimicrobial treatment, although surgical debridement of infected bite wounds was required in two cases. The betalactams and other families of antibiotics showed excellent in vitro activity against the five strains of Pasteurella isolated. CONCLUSIONS: PSB occurred in adult patients having a wide range of underlying illnesses and comorbidity factors. Most of them had contact with pets, though traumatic lesions were not present in all cases. Clinical presentation did not differ from other types of severe sepsis. Susceptibility and outcome of primary treatment with penicillins and other betalactams shows that they are still appropriate therapy. More emphasis should be placed on preventive measures related to care and hygiene among individuals with pets. PMID- 14525689 TI - [Epidemic outbreak of meningitis due to Echovirus type 13 on the island of Gran Canaria (Spain)]. AB - INTRODUCTION: Since the implementation of routine vaccination against mumps, enteroviruses have become the commonest cause of viral meningitis. In Spain the most frequently isolated echoviruses are serotypes 30, 9, 6 and 4.Objectives. Determine the clinical-epidemiological characteristics of a meningitis outbreak due to echovirus type 13. METHODS: We studied 152 cases of enteroviral meningitis diagnosed in the year 2000. Viral isolation was performed on fibroblast MRC-5 and human rhabdomyosarcoma cells. The viruses were identified with monoclonal antibodies and serotyped by neutralization. RESULTS: Echovirus was isolated in 131 out of 152 CSF samples (86.2%). In 21 patients with negative CSF culture, the diagnosis was made by echovirus isolation from upper respiratory tract and/or feces samples. The cytopathic effect was seen in all cases in monolayers of rhabdomyosarcoma cells. The average age of the patients was 67 months (range: 1 350) and the male:female ratio was 2:1. Most of the patients had fever, headache and other signs of meningitis. Hospitalization was required in 52.6%. All patients were discharged without sequelae. The highest incidence was recorded from April to June. CONCLUSIONS: There had been no record of the presence of echovirus 13 in Spain in recent years until the emergence of this outbreak. Isolation of the virus from sites other than CSF is often helpful in establishing the diagnosis and patient management. Enterovirus isolates must be specifically serotyped to determine enteroviral activity in the population, which can be sporadic or epidemic. PMID- 14525690 TI - [Comparative study of three culture media for detecting group B Streptococcus colonization in pregnant women]. AB - OBJECTIVE: To compare the efficacy of three culture media to detect vaginal and rectal colonization by group B streptococci (GBS) in pregnant women. METHODS: We processed 1334 samples from 861 pregnant women: in 388 patients only vaginal swab was cultured (Group A) and in 473 vaginal and rectal swabs were cultured (Group B). Samples were inoculated on blood agar with colistin-nalidixic acid (BA-CNA), on plates with Granada medium, and on Todd-Hewitt broth (THB) with amikacin followed by subculture in BA-CNA. RESULTS: GBS was isolated in 181 samples (13.6%): 114 vaginal swabs (13.2%) and 67 rectal swabs (14.2%). Among the positive samples, GBS was isolated on BA-CNA in 60.5% of vaginal swabs and in 59.7% of rectal swabs, on Granada medium in 80.7% of vaginal swabs and in 91% of rectal swabs, and on THB in 97.4% of vaginal swabs and in 97% of rectal swabs.We detected 130 GBS carriers, 54 (13.9%) in Group A and 76 (16.1%) in Group B. The percent of carriers detected in groups A and B, respectively, was 59.3% and 75% with BA-CNA, 77.8% and 93.4% with Granada medium, and 96.3% and 97.4% with THB. CONCLUSIONS: THB was the most reliable medium for the detection of GBS. Use of Granada medium allows fast detection of about 87% of carriers. The combination of THB and an initial plate of Granada medium or BA-CNA allows detection of more than 99% of GBS carriers. PMID- 14525691 TI - [Clinical management of community-acquired pneumonia]. PMID- 14525692 TI - [Eukaryotic antibiotic peptides: a new alternative in clinical practice?]. AB - Eukaryotic antibiotic peptides are key components of innate immunity. They act as a first barrier against invading pathogens. Expectations about their clinical use have been raised as they are active against a wide range of pathogens, and unlikely to induce resistance. Both characteristics are related to their lethal mechanism, based on interaction with anionic phospholipids in the outer facet of the cytoplasmic membrane of the pathogens, leading to permeabilization. This review will especially focus on antibiotic peptides of human origin. Their mechanism of action, strategies of pathogen resistance, and role in activities other than microbicidal effect will be described. Practical applications and prospects of future use in anti-infectious therapy will also be discussed. PMID- 14525694 TI - [Recurrent urticaria of two years' duration]. PMID- 14525693 TI - [Systemic antifungal agents]. AB - Invasive fungal infection is the infectious complication with highest associated mortality. Until the 90's amphotericin B was the only drug available to treat these infections. Its spectrum of antifungal activity is excellent, but its use is associated with toxicity in many cases. The development of amphotericin B lipid formulations has resulted in a significant decrease in most of the side effects associated with this drug. Triazoles are safe and effective for treating most invasive fungal infections. Fluconazole is an excellent drug for the prevention and treatment of Candida and Cryptococcus infections, itraconazole has good activity against Candida and Aspergillus, and voriconazole has shown to be better than amphotericin B for invasive aspergillosis. Caspofungin belongs to a new group of antifungal agents, the echinocandins, which are very safe and present excellent activity against Candida and Aspergillus. PMID- 14525695 TI - [Q fever and myocarditis]. PMID- 14525697 TI - [Recent increase in vaccine failures due to Haemophilus influenzae serotype b]. PMID- 14525696 TI - [Laboratory diagnosis of parotiditis in the post-vaccination era: new strategies]. PMID- 14525698 TI - [Candida folliculitis in intravenous drug users]. PMID- 14525699 TI - [Acute recurrent bacterial meningitis secondary to epidural catheter infection]. PMID- 14525700 TI - [Contribution of the microbiology laboratory to epidemiologic surveillance]. PMID- 14525701 TI - [Evaluation of in the impact factor of journals included in the Infectious Diseases category of the Journal Citation Report (1991-2001)]. PMID- 14525702 TI - [Legionella pneumophila serogroup 1]. PMID- 14525704 TI - [Intercontinental spread of a trimethoprim-resistant strain of Shigella flexneri]. AB - INTRODUCTION: In this study we characterize a trimethoprim-resistant strain of Shigella flexneri recovered from stool samples of an international traveler after a trip to Kenya, and analyze its epidemiological relationship with a set of strains having similar characteristics from Tanzania. METHODS: Clonal relationships were studied by three techniques - plasmid profile, repetitive element (REP)-PCR and pulse-field gel electrophoresis (PFGE). The presence of type 1 integrons was studied by PCR and sequencing. The location and transferability of the detected integron was analyzed by conjugation and PCR. RESULTS: The epidemiological studies showed that all six strains studied belonged to the same clone. Furthermore, all of them carried the same gene encoding for trimethoprim resistance (dfrA7), which was located in an integron within a chromosome. CONCLUSION: Continuous epidemiological surveillance is required to control the spread of pathogenic microorganisms and the dissemination of resistance-encoding genes among geographical areas. PMID- 14525703 TI - [Community-acquired pneumonia due to Legionella pneumophila serogroup 1. Study of 97 cases]. AB - INTRODUCTION: Legionella pneumophila is the causal agent of 5% to 12% of sporadic community-acquired pneumonia cases, though rates are changing with the use of new diagnostic methods. METHODS: This is a retrospective study of all patients admitted to our hospital with community-acquired pneumonia due to Legionella pneumophila between 1997 and 2001. Diagnostic criteria included either a positive Legionella serogroup 1 urinary antigen test or seroconversion and a chest radiograph consistent with pneumonia. RESULTS: A total of 97 patients were studied. Ninety cases (92.8%) were community-acquired and 7 (7.2%) were associated with travelling. In 82 cases (84.5%) the presentation was sporadic. Seventy-five patients were smokers (77.3%). The most common symptoms were fever in 91 patients (93.8%) and cough in 67 (68.1%). In five patients (5.2%) creatine phosphokinase concentrations were over 5 times their baseline values (in two over 100 times); four of these patients presented acute renal failure. Seroconversion was observed in 23/42 patients (54.8%). There were no statistically significant differences between the administration of erythromycin or clarithromycin in monotherapy, or in combination with rifampin. Nineteen patients (19.6%) presented acute renal failure and mechanical ventilation was necessary in 22 (22.7%). Twelve patients died (12.5%). Independent prognostic factors associated with death included respiratory rate > 30 breaths/min, urea > 60 mg/dL and PaO2 < 60 mmHg. A significant linear association was found between severity scale scores and the presence of complications or mortality. CONCLUSION: The Legionella urinary antigen test permits early diagnosis and treatment of this disease. The severity scale is an indicator of complications or death. PMID- 14525706 TI - [Respiratory infections outside the hospital. DIRA study]. AB - INTRODUCTION: Most visits to the primary care center are for infection and particularly respiratory tract infection. Antimicrobial administration for these clinical processes is common and these agents are often used to treat viral infections. La Fundacion para el Estudio de la Infeccion (Foundation for the Study of Infection) designed the DIRA (Dia de la Infeccion Respiratoria del Adulto, Adult Respiratory Infection Day) Project to investigate and assess the impact of this problem. METHODS: The study design consisted of one-day cross sections with the participation of 720 physicians belonging to Primary Health Care Centers from the 17 Autonomic Regions of Spain, establishing a one-day cross section every three months for one year. Epidemiologic, clinical and therapeutic factors were evaluated. RESULTS: The total number of visits attended was 72 929, and 14 426 patients had infectious processes (43.9%). Among these, 9145 (63.4%) had a respiratory infection. The mean age of the patients was 44.6 years and 34.1% had an underlying condition. Common cold was the most frequent diagnosis. Antibiotics were prescribed in 53.2% of patients. Results were compared to those of a previous study. DISCUSSION: Infection in general and respiratory infection in particular is frequently attended in primary care. Antibiotics are widely used in our setting and self-medication is common. PMID- 14525705 TI - [Comparative in vitro activity of garenoxacin (BMS-284756). Sentry program, Spain (1999-2000)]. AB - INTRODUCTION: To evaluate the in vitro activity of the new des-fluoro quinolone, garenoxacin (BMS-284756), compared to activities of ciprofloxacin, levofloxacin, and gatifloxacin in clinical isolates recovered over 1999 and 2000 within the SENTRY antimicrobial surveillance program. METHODS: Quinolone-MICs were performed using the standard NCCLS microdilution technique in 2599 isolates recovered from Hospital Ramon y Cajal (Madrid), Virgen Macarena (Sevilla), and Bellvitge (Barcelona). RESULTS: The modal MIC range value exhibited by garenoxacin ( < or = 0.03-0.12 mg/L) for Enterobacteriaceae was similar to that of the other quinolones tested. A total of 70% of Pseudomonas aeruginosa isolates were susceptible to garenoxacin and 85% to ciprofloxacin and levofloxacin. Garenoxacin exhibited the highest activity in Staphylococcus aureus, including both methicillin-susceptible and -resistant isolates, with MIC90 values of < or = 0.03 and 2 mg/L, respectively. All Streptococcus pneumoniae isolates were susceptible to garenoxacin, regardless of their penicillin susceptibility status; in terms of MIC90, garenoxacin was 16 times more active than ciprofloxacin and levofloxacin and 4-8 times more active than gatifloxacin. All 6 ciprofloxacin-resistant S. pneumoniae strains showed garenoxacin MIC values ranging from < or = 0.03 to 0.5 mg/L. In Haemophilus influenzae and Moraxella catarrhalis, garenoxacin displayed excellent in vitro activity (MIC < or = 0.06 mg/L), similar to that of the other quinolones tested. CONCLUSIONS: Garenoxacin activity was similar to the activity of other quinolones in Enterobacteriaceae, but was lower in P. aeruginosa. Garenoxacin activity was clearly higher than that of other quinolones in gram positive isolates, including methicillin-resistant S. aureus and S. pneumoniae with reduced penicillin susceptibility. PMID- 14525707 TI - [Prevention of perinatal group B streptococcal disease. Spanish revised guidelines. The Spanish Society of Obstetrics and Gynecology, The Spanish Society of Neonatology, The Spanish Society of Infectious Diseases and Clinical Microbiology, The Spanish Society of Chemotherapy, and the The Spanish Society of Family and Community Medicine]. PMID- 14525708 TI - [Human herpesvirus type 6 and type 7 in transplant recipients]. AB - Recent years have witnessed a growing interest in the role of human herpesvirus (HHV) type 6 and type 7 as emerging pathogens or copathogens in transplant recipients. Both HHV-6 and HHV-7 belong to the beta-herpesvirus family and are closely related to another member of the family, cytomegalovirus. After the primary infection, these viruses remain latent in the human host and can reactivate after transplantation. Various clinical processes such as fever, rash, pneumonitis, encephalitis, hepatitis, and myelosuppression have been described in association with herpesvirus. Moreover, a growing body of evidence suggests that the major impact of HHV-6 and HHV-7 reactivation in transplantation is related to indirect effects, such as their association with cytomegalovirus disease, increased opportunistic infections, and graft dysfunction and rejection. The pathogenesis of HHV-6 and HHV-7 during the post-transplantation period, the methods used for their diagnosis, and the evaluation of antiviral drugs and strategies for their prevention and treatment are now the subject of extensive research. PMID- 14525709 TI - [Characteristics of antiviral drugs]. AB - This article summarizes the principal characteristics of the drugs used to treat viral infections, with the exception of human immunodeficiency virus infection. It includes antiviral agents active against herpes virus, cytomegalovirus, hepatitis B and C virus, and respiratory viruses, such as influenza and respiratory syncytial virus. Dosage according to the indication, dose adjustment in the case of renal or hepatic insufficiency, significant pharmacokinetic characteristics, and the main adverse effects and interactions are described. PMID- 14525710 TI - [Progressive lung disease in an HIV-negative patient]. PMID- 14525712 TI - [Evaluation of the ROBOBACT system for automatic processing of stool cultures]. PMID- 14525711 TI - [Recurrent pancreatitis after cholecystectomy]. PMID- 14525713 TI - [Non-specific proctitis in a patient with HIV infection]. PMID- 14525714 TI - [Acute calculous cholecystitis associated with Brucella melitensis]. PMID- 14525715 TI - [Acute Staphylococcus lugdunensis endocarditis with septic cerebral and pulmonary emboli, showing favorable evolution]. PMID- 14525716 TI - Comparing multifactorial lifestyle interventions and stress management in coronary risk reduction. AB - The aim of this study was to compare the effects of residential multifactorial cardiac rehabilitation, outpatient multifactorial rehabilitation, stress management, and standard coronary rehabilitation, on cardiac risk reduction. Out of 144 eligible male patients recently treated with percantaneous transluminal coronary angiography (PTCA), coronary artery bypass graft (CABG), or acute myocardial infarction (AMI), 132 were randomized into this study. All interventions covered a 12-month active intervention, intense during the first months and subsequently leveled out. Main assessments were performed before randomization and after the intervention. Patients offered behavioral rehabilitation showed improved self-reported healthy diet habits and exercise frequency, and higher internal locus of control. Although blood lipids, exercise capacity, body mass, anxiety, depression, and Type A scores were changed in the expected direction, no significant difference emerged between active intervention and the standard care condition. Standard care of today appears to have great potential in particular if supplemented with some kind of stress management. PMID- 14525717 TI - Perceived stress as a predictor of the self-reported new diagnosis of symptomatic CHD in older women. AB - This article describes one aspect of a prospective cohort study of 10,432 women aged between 70 and 75 years. After a 3-year period, 503 women self-reported a new diagnosis by a doctor of angina or myocardial infarction (symptomatic coronary heart disease [CHD]). Time one psychosocial variables (Duke Social Support Index, time pressure, Perceived Stress Scale, Mental Health Index, having a partner, educational attainment, and location of residence) were analyzed using univariate binary logistic regression for their ability to predict subsequent symptomatic CHD. Of these variables, the Duke Social Support Index, Perceived Stress Scale and the Mental Health Index were found to be significant predictors of symptomatic CHD diagnosis. Only the Perceived Stress Scale, however, proved to be a significant independent predictor. After controlling for time one nonpsychosocial variables, as well as the frequency of family doctor visits, perceived stress remained a significant predictor of the new diagnosis of symptomatic CHD in this cohort of older women over a 3-year period. PMID- 14525718 TI - Associations between repression, general maladjustment, body weight, and body shape in older males: the Normative Aging Study. AB - We examined relationships between repression, general maladjustment, body mass index (BMI), and waist-to-hip ratio (WHR). The participants were 1,081 healthy older men from the Normative Aging Study. Repression and General Maladjustment Scales of the Minnesota Multiphasic Personality Inventory were composite measures of personality. Repression was associated with lower BMI and WHR, and maladjustment with higher BMI and WHR. However, associations between WHR and personality dimensions were no longer significant when controlling for BMI, but associations between BMI and personality dimensions remained significant when controlling for WHR. These effects were explained by differing relationships between WHR, repression, and maladjustment for normal weight, overweight, and obese individuals. Specifically, associations between repression, maladjustment, and body shape were significant for normal weight and overweight individuals, but not for obese individuals. Health behaviors including smoking did not mediate relationships between repression, maladjustment, and body shape, but might be considered in future studies as mechanisms underlying links between personality and body shape. PMID- 14525719 TI - Associations among social support, income, and symptoms of depression in an educated sample: the UNC Alumni Heart Study. AB - It has been suggested that the inverse association between social support and depression may be stronger in persons with lower income. This study tested the support x income hypothesis in a sample of 2,472 individuals enrolled in the UNC Alumni Heart Study. The income was examined as a moderator of the relation between support and self-reported ratings of symptoms of depression. The appraisal subscale of the Interpersonal Support Evaluation Scale, household income level, and their interaction were modeled as predictors of depression ratings. The support x income interaction term was significantly associated with symptoms of depression, F (2, 2471)=4.71, p=.007. Social support was more strongly associated inversely with depression ratings in persons with lower income as compared to those with higher income. The present results extend previous work regarding the moderating effect of income to a sample of relatively high education and income level. PMID- 14525720 TI - Adverse experience in childhood as a developmental risk factor for altered immune status in adulthood. AB - Compelling evidence is now available that adverse childhood experiences are associated with adult pathology. However, understanding of the pathways and mechanisms underlying these associations is limited. Participants in the European Prospective Investigation into Cancer and Nutrition in Norfolk, UK (EPIC Norfolk), aged 40 to 80 years, provided an opportunity to investigate the hypothesis that adverse experience in childhood is associated with peripheral leukocyte count in adulthood in the context of a large-scale population-based cohort study. White blood cell counts were available from 11,367 participants and, after a mean interval of 44 months, from 11,857 at a second health check. A self-completion questionnaire that included the assessment of adverse experience during childhood was administered during the interval between health checks. Associations were observed between early adverse experiences and lymphocyte counts at both health checks. Lifestyle factors accounted for about half of this association. Caution is needed in the interpretation of these findings that require replication but they may be seen to aid understanding of the mechanisms through which early environmental exposures act. PMID- 14525721 TI - Use of hormone replacement therapy among Danish nurses at increased risk of osteoporosis. AB - Observational studies and recent randomized trials have shown that postmenopausal hormone replacement therapy (HRT) may reduce the risk of osteoporotic fractures by about 30 to 40%. In this study we used a log linear graphical model to determine whether women with a known increased risk of osteoporosis were more likely to use HRT than other women and to examine whether women at increased risk modified this risk through their lifestyle. Cox regression analysis was used to analyze if women at risk of osteoporosis used HRT longer than women not at risk. Participants were Danish female nurses who, in 1993, were between 50 and 69 years of age (N=14,865). Data were collected from postal questionnaires. We concluded that nurses with a known family history of osteoporosis more often used HRT than nurses without this risk. No other direct associations were found between biological risk factors and ever use of HRT. The presence of biological risk factors of osteoporosis was not consistently modified by a healthier lifestyle. Nurses with a low body mass index (BMI) with a known family history of osteoporosis continued to use HRT longer than nurses without these risk factors. PMID- 14525722 TI - Postnatal hypoxic-ischemic brain injury alters mechanisms mediating neuronal glucose transport. AB - We examined the effect of hypoxic ischemia and hypoxia vs. normoxia on postnatal murine brain substrate transporter concentrations and function. We detected a transient increase in the neuronal brain glucose transporter isoform (GLUT-3) in response to hypoxic ischemia after 4 h of reoxygenation. This increase was associated with no change in GLUT-1 (blood-brain barrier/glial isoform), monocarboxylate transporter isoforms 1 and 2, synapsin I (neuronal marker), or Bax (proapoptotic protein) but with a modest increase in Bcl-2 (antiapoptotic mitochondrial protein) protein concentrations. At 24 h of reoxygenation, the increase in GLUT-3 disappeared but was associated with a decline in Bcl-2 protein concentrations and the Bcl2:Bax ratio, an increase in caspase-3 enzyme activity (apoptotic effector enzyme), and extensive DNA fragmentation, which persisted later in time (48 h) only in the hippocampus. Hypoxia alone in the absence of ischemia was associated with a transient but modest increase in GLUT-3 and synapsin I protein concentrations, which did not cause significant apoptosis and/or necrosis. Assessment of glucose transporter function by 2-deoxyglucose (2 DG) uptake using two distinct techniques, namely positron emission tomography (PET) and the modified Sokoloff method, revealed a discrepancy due to glucose uptake by extracranial Harderian glands that masked the accurate detection of intracranial brain glucose uptake by PET scanning. The modified Sokoloff method assessing 2-DG uptake revealed that the transient increase in GLUT-3 was critical in protecting against a decline in brain glucose uptake. We conclude that hypoxic ischemic brain injury is associated with transient compensatory changes targeted at protecting glucose delivery to fuel cellular energy metabolism, which then may delay the processes of apoptosis and cell necrosis. PMID- 14525723 TI - Heterogeneous patterns of pH regulation in glial cells in the dorsal and ventral medulla. AB - We examined pH regulation in two chemosensitive areas of the brain, the retrotrapezoid nucleus (RTN) and the nucleus tractus solitarius (NTS), to identify the proton transporters involved in regulation of intracellular pH (pHi) in medullary glia. Transverse brain slices from young rats [postnatal day 8 (P8) to P20] were loaded with the pH-sensitive probe 2',7'-bis (2-carboxyethyl)-5,6 carboxyfluorescein after kainic acid treatment removed neurons. Cells were alkalinized when they were depolarized (extracellular K+ increased from 6.24 to 21.24 mM) in the RTN but not in the NTS. This alkaline shift was inhibited by 0.5 mM DIDS. Removal of CO2/HCO3- or Na+ from the perfusate acidified the glial cells, but the acidification after Na+ removal was greater in the RTN than in the NTS. Treatment of the slice with 5-(N-ethyl-N-isopropyl)amiloride (100 microM) in saline containing CO2/HCO3- acidified the cells in both nuclei, but the acidification was greater in the NTS. Restoration of extracellular Cl- after Cl- depletion during the control condition acidified the cells. Immunohistochemical studies of glial fibrillary acid protein demonstrated much denser staining in the RTN compared with the NTS. We conclude that there is evidence of Na+-HCO3- cotransport and Na+/H+ exchange in glia in the RTN and NTS, but the distribution of glia and the distribution of these pH-regulatory functions are not identical in the NTS and RTN. The differential strength of glial pH regulatory function in the RTN and NTS may also alter CO2 chemosensory neuronal function at these two chemosensitive sites in the brain stem. PMID- 14525724 TI - Maturation of the homeothermic response of heart rate to altered ambient temperature in developing chick hatchlings (Gallus gallus domesticus). AB - On the basis of evidence showing that instantaneous heart rate (IHR) of chick hatchlings responds to exposure to altered ambient temperature (Ta; Tazawa H, Moriya K, Tamura A, and Akiyama R. Comp Biochem Physiol A 131A: 797-803, 2002), we elucidate here the developmental timeline for the homeothermic response of HR in newly hatched chicks (days 0-7) maintained at room temperature ( approximately 24-27 degrees C). Hatchlings were exposed to Ta of 25, 35, and 25 degrees C for 1 h periods, respectively, and IHR was measured together with skin temperature (Ts) during this warming and cooling bout. Early 0-day-old (0 day) chicks responded to warming and cooling exposures with various changes in HR baseline. In newly hatched chicks (0-7 h old), HR baseline was elevated during warming (Delta126 beats/min, n = 13) and declined during cooling (-Delta94 beats/min). With progress of development on day 0, the elevation of HR baseline during warming decreased and advanced 0-day chicks tended to decrease HR baseline during warming rather than increase HR. The more developed 1- to 7-day-old chicks exhibited the expected homeothermic decrease in HR during warming. The diurnal variations of HR responses during warming and cooling on the first day of post-egg life indicate that pronounced development of thermoregulatory competence occurs during the day of hatching (day 0). The response of IHR fluctuations to altered Ta was observed in the form of low- and high-frequency oscillations. High-frequency oscillations corresponding to respiratory sinus arrhythmia developed as the hatchlings aged. There was a significant increase in the number of chicks exhibiting both low- and high-frequency oscillations that depended on age and the development of thermoregulatory competence of hatchlings. PMID- 14525725 TI - Autologous fat transplants influence compensatory white adipose tissue mass increases after lipectomy. AB - Direct tests of the hypothesized total body fat regulatory system have been accomplished by partial surgical lipectomy. This usually results in the restoration of the lipid deficit through compensatory increases in nonexcised white adipose tissue (WAT) masses of ground squirrels, laboratory rats, and mice, as well as Siberian and Syrian hamsters. We challenged this hypothesized total body fat regulatory system by testing the response of Siberian hamsters to 1) lipid deficits [lipectomy; primarily bilateral epididymal WAT (EWAT) removal], 2) lipid surfeits (addition of donor EWAT with no lipectomy), 3) no net change in lipid [EWAT or inguinal WAT (IWAT) lipectomy with the excised fat replaced to a new location (autologous)], 4) lipectomy with the same pad (EWAT lipectomy only) added from a sibling (nonautologous), and 5) sham surgeries for each treatment. Food intake generally was not affected. Body mass was not affected across all treatments. Grafts approximately 3 mo later had normal appearance both macro- and microscopically and were revascularized. The normal lipectomy-induced compensatory increases in nonexcised WAT masses surprisingly were exaggerated with autologous EWAT transplants, but not for autologous IWAT or nonautologous EWAT transplants. There was no compensatory decrease in native WAT masses with nonautologous EWAT additions. Collectively, only lipectomy triggered reparation of the lipid deficit, but the other manipulations did not, suggesting a system biased toward rectifying decreases in lipid or an inability of the hypothesized total body fat regulatory system to recognize WAT transplants. PMID- 14525726 TI - NO, but not CO, attenuates anaphylaxis-induced postsinusoidal contraction and congestion in guinea pig liver. AB - The pathophysiology of the hepatic vascular response to anaphylaxis in guinea pig is not known. We studied effects of anaphylaxis on hepatic vascular resistances and liver weight in isolated perfused livers derived from guinea pigs sensitized with ovalbumin. We also determined whether nitric oxide (NO) or carbon monoxide (CO) modulates the hepatic anaphylaxis. The livers were perfused portally and recirculatingly at constant flow with diluted blood. With the use of the double occlusion technique to estimate the hepatic sinusoidal pressure (Pdo), portal venous resistance (Rpv) and hepatic venous resistance (Rhv) were calculated. An antigen injection caused venoconstriction characterized by an increase in Rpv greater than Rhv and was accompanied by a large liver weight gain. Pretreatment with the NO synthase inhibitor NG-nitro-l-arginine methyl ester, but not the heme oxygenase inhibitor zinc protoporphyrin IX, potentiated the antigen-induced venoconstriction by increasing both Rpv and Rhv (2.2- and 1.2-fold increase, respectively). In conclusion, anaphylaxis causes both pre- and postsinusoidal constriction in isolated guinea pig livers. However, the increases in postsinusoidal resistance and Pdo cause hepatic congestion. Endogenously produced NO, but not CO, modulates these responses. PMID- 14525727 TI - Regulation of butyrate uptake in Caco-2 cells by phorbol 12-myristate 13-acetate. AB - Butyrate and the other short-chain fatty acids (SCFAs) are the most abundant anions in the colonic lumen. Also, butyrate is the preferred energy source for colonocytes and has been shown to regulate colonic electrolyte and fluid absorption. Previous studies from our group have demonstrated that the HCO(3)( )/SCFA(-) anion exchange process is one of the major mechanisms of butyrate transport across the purified human colonic apical membrane vesicles and the apical membrane of human colonic adenocarcinoma cell line Caco-2 and have suggested that it is mainly mediated via monocarboxylate transporter-1 (MCT-1) isoform. However, little is known regarding the regulation of SCFA transport by various hormones and signal transduction pathways. Therefore, the present studies were undertaken to examine whether hydrocortisone and phorbol 12-myristate 13 acetate (PMA) are involved in a possible regulation of the butyrate/anion exchange process in Caco-2 cells. The butyrate/anion exchange process was assessed by measuring a pH-driven [(14)C]butyrate uptake in Caco-2 cells. Our results demonstrated that 24-h incubation with PMA (1 microM) significantly increased [(14)C]butyrate uptake compared with incubation with 4alphaPMA (inactive form). In contrast, incubation with hydrocortisone had no significant effect on butyrate uptake in Caco-2 cells compared with vehicle (ethanol) alone. Induction of butyrate uptake by PMA appeared to be via an increase in the maximum velocity (V(max)) of the transport process with no significant changes in the K(m) of the transporter for butyrate. Parallel to the increase in the V(max) of [(14)C]butyrate uptake, the MCT-1 protein level was also increased in response to PMA incubation. Our studies demonstrated that the butyrate/anion exchange was increased in response to PMA treatment along with the induction in the level of MCT-1 expression in Caco-2 cells. PMID- 14525728 TI - Gastric ulcers reduce A-type potassium currents in rat gastric sensory ganglion neurons. AB - Voltage-dependent potassium currents are important contributors to neuron excitability and thus also to hypersensitivity after tissue insult. We hypothesized that gastric ulcers would alter K(+) current properties in primary sensory neurons. The rat stomach was surgically exposed, and a retrograde tracer (1,1'-dioctadecyl-3,3,3,3'-tetramethylindocarbocyanine methanesulfonate) was injected into multiple sites in the stomach wall. Inflammation and ulcers were produced by 10 injections of 20% acetic acid (HAc) in the gastric wall. Saline (Sal) injections served as control. Nodose or T9-10 dorsal root ganglia (DRG) cells were harvested and cultured 7 days later to record whole cell K(+) currents. Gastric sensory neurons expressed transient and sustained outward currents. Gastric inflammation significantly decreased the A-type K(+) current density in DRG and nodose neurons (Sal vs. HAc-DRG: 82.9 +/- 7.9 vs. 46.5 +/- 6.1 pA/pF; nodose: 149.2 +/- 10.9 vs. 71.4 +/- 11.8 pA/pF), whereas the sustained current was not altered. In addition, there was a significant shift in the steady state inactivation to more hyperpolarized potentials in nodose neurons (Sal vs. HAc: -76.3 +/- 1.0 vs. -83.6 +/- 2.2 mV) associated with an acceleration of inactivation kinetics. These data suggest that a reduction in K(+) currents contributes, in part, to increased neuron excitability that may lead to development of dyspeptic symptoms. PMID- 14525729 TI - TNFR1 mediates the radioprotective effects of lipopolysaccharide in the mouse intestine. AB - LPS is radioprotective in the mouse small intestine through a mechanism that includes the synthesis of cyclooxygenase-2 (COX-2) and PGE2. The goal of this study was to identify the intermediate steps in this process. We used wild-type (WT) C57BL/6 mice and knockouts for tumor necrosis factor receptors 1 and 2 (TNFR1-/-, TNFR2-/-) and recombination-activating gene 1-/- mice. Mice were given parenteral LPS and then subjected to 12 Gy total body gamma irradiation. The number of surviving intestinal crypts was assessed 3.5 days after irradiation using a clonogenic assay. Crypt cell apoptosis was assessed by histology. Parenteral administration of LPS induced COX-2 expression, PGE2 production, and radioprotection in WT and TNFR2-/- mice but not in TNFR1-/- mice. TNFR1-/- mice were radioprotected by administration of exogenous 16,16-dimethyl PGE2. Immunohistochemical studies localized TNFR1 and COX-2 expression to subeptihelial fibroblasts and villus epithelial cells. Radiation-induced apoptosis was reduced by pretreatment with LPS in WT and TNFR2-/- mice but not in TNFR1-/- mice. In the absence of LPS, crypt survival was elevated in TNFR1-/- when compared with WT mice. These findings demonstrate that TNFR1 function is required for LPS-induced radioprotection in C57BL/6 mice and define an essential role for TNFR1 function in the induction of COX-2 expression and PGE2 production in this process. The immunolocalization of TNFR1 and COX-2 expression to subepithelial fibroblasts following LPS administration suggests that this cell type plays an intermediate role in LPS-induced radioprotection in the intestine. PMID- 14525730 TI - Prevention of descending pneumonia in rts with perflubron-delivered tobramycin. AB - OBJECTIVES: Patients undergoing emergent endotracheal intubation are at increased risk for developing pneumonia. Although numerous strategies have been investigated to reduce ventilator-associated pneumonia (VAP), the incidence of VAP and its associated mortality remains high. This investigation tested the hypothesis that LiquiVent (Alliance Pharmaceutical, San Diego, CA-LV) delivered antibiotics (via spray-dried microspheres-SDM) would improve survival in a rat model of descending gram-negative pneumonia. METHODS: Wistar rats (n = 49) were randomized to receive prophylaxis with 1). nothing (controls); 2). intramuscular (IM) tobramycin, 3). intratracheal LV plus SDM shells (vehicle), 4). intratracheal LV plus SDM shells plus IM tobramycin, or 5). intratracheal LV plus SDM containing 1 mg/kg of tobramycin. All interventions were given 24 hours before a bacterial challenge with 10(8) colony-forming units of intratracheal Klebsiella pneumoniae. Mortality at ten days was the sole outcome measure. Survival in individual groups was compared with controls by Fisher's exact test with Bonferroni correction for multiple comparisons. RESULTS: All animals in the control group died of pneumonia within ten days of bacterial inoculation (0% survival). Prophylaxis with either IM tobramycin or SDM vehicle plus IM tobramycin provided no protection (0% survival). This is in sharp contrast to the cohort receiving pretreatment with tobramycin-containing SDM delivered via LV, in which 60% of the animals survived to study completion (p < 0.05). CONCLUSIONS: Prophylaxis with SDM containing antibiotics delivered in low-dose LV provided significant protection in a rat model of descending gram-negative pneumonia. These data support the hypothesis that perfluorocarbon-delivered intratracheal antimicrobials may be useful in the prevention of VAP. PMID- 14525732 TI - Mitigation of pennyroyal oil hepatotoxicity in the mouse. AB - OBJECTIVES: Pennyroyal oil ingestion has been associated with severe hepatotoxicity and death. The primary constituent, R-(+)-pulegone, is metabolized via hepatic cytochrome P450 to toxic intermediates. The purpose of this study was to assess the ability of the specific cytochrome P450 inhibitors disulfiram and cimetidine to mitigate hepatotoxicity in mice exposed to toxic levels of R-(+) pulegone. METHODS: 20-g female BALB/c mice were pretreated with either 150 mg/kg of cimetidine intraperitoneal (IP), 100 mg/kg of disulfiram IP, or both. After one hour, mice were administered 300 mg/kg of pulegone IP and were killed 24 hours later. Data were analyzed using ANOVA. Post-hoc t-tests used Bonferroni correction. RESULTS: There was a tendency for lower serum glutamate pyruvate transaminase in the disulfiram and cimetidine groups compared with the R-(+) pulegone group. The differences were significant for both the cimetidine and the combined disulfram and cimetidine groups compared with the R-(+)-pulegone group. Pretreatment with the combination of disulfiram and cimetidine most effectively mitigated R-(+)-pulegone-induced hepatotoxicity. CONCLUSIONS: Within the limitations of a pretreatment animal model, the combination of cimetidine and disulfiram significantly mitigates the effects of pennyroyal toxicity and does so more effectively than either agent alone. These data suggest that R-(+)-pulegone metabolism through CYP1A2 appears to be more important in the development of a hepatotoxic metabolite than does metabolism via CYP2E1. PMID- 14525733 TI - Development of a porcine excisional wound model. AB - OBJECTIVES: To develop a porcine model for cutaneous tangential excisional wounds. METHODS: This was a prospective, longitudinal, blinded experiment on four isoflurane-anesthetized swine. Forty standardized excisional wounds (1 square inch each) were created with an electric dermatome set at a depth of 300, 600, 750, or 900 microm. Full-thickness wounds were created with a surgical blade (n = 8 for each depth). Wounds were treated with polyurethane film. Full-thickness punch biopsies were taken after one, two, three, four, five, and 14 days for blinded histopathological evaluation. The main outcome was the percent wound reepithelialization (REP) calculated by dividing the length of neoepidermis in cross-section by the specimen's diameter (interobserver correlation = 0.99). Analysis of variance was used to compare percent REP among wound depths. This study had 80% power to detect a 33% difference in REP across groups (two-tailed alpha = 0.05). RESULTS: There were significant differences in percent REP across wound depths for days 2-5 (p < 0.001 for all days). The more superficial wounds (300 to 600 microm) were mostly reepithelialized by day 4, whereas wounds ranging in depth from 750 to 900 micro m were mostly healed by day 5. Full-thickness wounds had no evidence of REP even after 14 days. CONCLUSIONS: The authors describe an excisional wound model in swine. Rapid reepithelialization occurs within three to five days in wounds <900 microm deep. PMID- 14525734 TI - Neurologic outcome score for infants and children. AB - OBJECTIVES: To develop and validate a practical outcome instrument applicable to a broad range of neurologic deficits in children. METHODS: Reliability testing of a draft version of the Neurologic Outcome Scale for Infants and Children (NOSIC) in 100 children with a wide range of ages and levels of neurologic function was performed. After review of the reliability data by a panel of experts, the NOSIC was revised. Validity and reliability testing of the final NOSIC was performed in a new population of 157 children, 52 with cerebral palsy, motor delay, or language delay. Interrater reliability was assessed using Spearman rank correlation coefficients of two investigators' scores. NOSIC scores were correlated with scores on criterion-standard neuropsychological tests to assess validity. RESULTS: The median NOSIC score for normal children was 98, interquartile range 96-100; the median score for abnormal children was 87, interquartile range 58-96. Interrater reliability of the NOSIC scores of 84 patients rated by both raters demonstrated excellent reliability (rho = 0.77, 95% confidence interval [CI] = 0.62 to 0.88). Correlation of the NOSIC scores of the 127 patients who had neuropsychological testing with applicable criterion standards was rho = 0.63, 95% CI = 0.50 to 0.74. CONCLUSIONS: The NOSIC is a practical, reliable, valid, instrument applicable to infants and children with a broad range of neurologic deficits. It should be a useful research tool when neurologic function is an important outcome measure. PMID- 14525735 TI - Comparison of the rapid emergency medicine score and APACHE II in nonsurgical emergency department patients. AB - OBJECTIVES: To improve the Rapid Acute Physiology Score (RAPS) as a predictor of in-hospital mortality in the nonsurgical emergency department (ED) by including age and oxygen saturation, and to compare this new system, Rapid Emergency Medicine Score (REMS), with the Acute Physiology and Chronic Health Examination (APACHE II) with reference to predictive accuracy. METHODS: This was a prospective cohort study. One hundred sixty-two critically ill patients consecutively admitted to the intensive care unit (ICU) during the period of one year, and 865 nonsurgical patients presenting to an adult emergency department (ED) and admitted to a medical department of a 1200-bed university hospital during two months, were enrolled. For all entries to the ED, RAPS was calculated and developed to include noninvasive peripheral oxygen saturation and patient age (REMS), as well as laboratory tests (APACHE II). These scores were calculated for each patient. RESULTS: REMS was found to be superior to RAPS in predicting in hospital mortality both in the critically ill patients admitted to the ICU and in the total sample (area under receiver-operating characteristic curve [AUC] 0.910 +/- 0.015 for REMS compared with 0.872 +/- 0.022 for RAPS, p < 0.001). An increase of 1 point in the 26-point REMS scale was associated with an odds ratio of 1.40 for in-hospital death (95% confidence interval = 1.36 to 1.45, p < 0.0001). The more advanced APACHE II was not found to be superior to REMS (AUC: 0.901 +/- 0.015, p = 0.218). CONCLUSIONS: RAPS could be improved as a predictor of in-hospital mortality in the nonsurgical ED by including oxygen saturation and patient age to the system. This new scoring system, REMS, had the same predictive accuracy as the well-established, but more complicated, APACHE II. PMID- 14525736 TI - General competencies are intrinsic to emergency medicine training: a multicenter study. AB - OBJECTIVES: The Accreditation Council for Graduate Medical Education (ACGME) has promulgated six areas called General Competencies (GCs) that residency programs are required to evaluate. The authors sought to determine if these domains were an intrinsic part of emergency medicine (EM) residency training by using a global assessment evaluation device. METHODS: This was an observational, multicenter, cross-sectional study that compared GC acquisition between first-, second-, and third-year (EM1, EM2, and EM3) residents. Five postgraduate year (PGY) 1 to PGY 3 allopathic EM programs in Michigan participated. A global assessment form using a 1 through 9 ordinal scale with 86 scoring items was given to program directors for each resident in their programs. Analysis of variance (ANOVA) was used to compare the means between EM1, EM2, and EM3 scores. RESULTS: Five EM programs evaluated 150 residents. The GC scores were as follows: Patient Care: EM1 4.92, EM2 5.79, and EM3 6.40; Medical Knowledge: EM1 4.90, EM2 5.80, and EM3 6.46; Practice-based Learning and Improvement: EM1 4.60, EM2 5.48, and EM3 6.16; Interpersonal and Communication Skills: EM1 4.99, EM2 5.39, and EM3 6.01; Professionalism: EM1 5.43, EM2 5.68, and EM3 6.27; Systems-based Practice: EM1 4.80, EM2 5.48, and EM3 6.21. ANOVA showed statistically significant differences (p < 0.001) for all GCs. CONCLUSIONS: EM residents from several residency programs showed statistically significant progressive acquisition of the ACGME GCs using a global assessment device. This suggests that the GCs may be an intrinsic component in the training of EM residents. PMID- 14525737 TI - Focused training of emergency medicine residents in goal-directed echocardiography: a prospective study. AB - OBJECTIVES: To determine if a focused transthoracic echocardiography (TTE) training course would improve the accuracy of completion and interpretation of a goal-directed TTE by emergency medicine residents. METHODS: This was a prospective, observational, educational study of the impact of a focused training course on the change in physician performance on pre- and postcourse examinations testing competency in goal-directed TTE defined by five criteria: 1). image orientation, 2). anatomy identification, 3). chamber size grading, 4). ventricular function estimation, and 5). pericardial effusion identification. Subjects included were emergency medicine residents with between ten and 20 hours of noncardiac ultrasound didactics and between 20 and >150 proctored noncardiac ultrasound examinations. All underwent five hours of focused echocardiography didactics and one hour of proctored practical echocardiography training designed and implemented by an emergency physician ultrasound director and a cardiologist. Before the start of the training course, participants completed two examinations: 1) written 23-question test on the above concepts and 2) performance of a TTE on a healthy subject testing 16 elements that define a properly performed examination. After the training course, participants again completed both examinations. RESULTS: A total of 21 emergency medicine residents qualified for and underwent standardized testing and training. The percentage correct on the precourse written examination was 54% (95% CI = 50% to 59%), and the postcourse examination score was 76% (95% CI = 71% to 80%) (p < 0.005, paired t-test). The percentage correct on the precourse practical examination was 56% (95% CI = 51% to 60%), and the postcourse examination score was 94% (95% CI = 91% to 96%) (p < 0.005). CONCLUSIONS: A focused six-hour echocardiography training course significantly improved emergency medicine residents' percentage scores on both written and practical examinations testing essential components required for correct goal-directed TTE performance and interpretation. PMID- 14525738 TI - The overcrowded emergency department: a comparison of staff perceptions. AB - OBJECTIVES: To study perceptions of physicians and nursing staff about real-time demands and capacity of an emergency department (ED). To use ED data to calculate proposed demand ratios called Real-time Emergency Analysis of Demand Indicators (READI) scores. To compare the READI scores with ED staff perceptions of demand and capacity. METHODS: This prospective study used a computerized clinical management system to provide data about ED demand and capacity. Physicians and staff charge nurses were surveyed about perceptions of ED demand and capacity. Results were compared with mathematical READI scores, which are proposed to objectively assess ED demand. Kappa scores were used to measure intrarater reliability between the physicians' and charge nurses' assessment of demand and between the staff assessments and the READI scores. RESULTS: Kappa scores of the perception of excess demand were as follows: between two physician groups, kappa = 0.392; between one physician group and charge nurses, kappa = 0.453; and between a second physician group and charge nurses, kappa = 0.243. Comparing respondents who indicated that demand had or had not exceeded capacity, one of the READI ratios, the Bed Ratio, showed a significant difference in mean, 0.245 (95% confidence interval = 0.153 to 0.336), between groups. CONCLUSIONS: Real time data may be used to predict ED demand and resource needs. Subjective assessment of excess ED demand did not correlate between physician groups or between physicians and charge nurses. Although there was a trend toward predicting excess demand with one of the READI scores, these scores did not correlate to staff perceptions. PMID- 14525739 TI - Panic disorder and emergency services utilization. AB - OBJECTIVES: The characteristics of patients with panic disorder in emergency department (ED) patient populations are unknown. This study compares demographic information and emergency care use among patients identified as having a high likelihood of having panic disorder with that of patients who tested negative on the screening test for panic disorder. METHODS: Prospective cross-sectional study of a convenience sample of patients presenting to an urban ED. Patients were excluded if they were aged 18 years or younger, were unstable, or could not speak English or Spanish. Of 968 patients, 813 agreed to participate. Over a period of 23 days, patients were administered a Diagnostic and Statistical Manual (DSM)-IV screening questionnaire (PRIME-MD) for panic disorder along with a survey assessing their use of medical services during the prior year. RESULTS: One hundred patients (12.3%) met PRIME-MD criteria for having a high likelihood of panic disorder. Patients with Medicare were 2.84 times more likely to have a positive result on the screening test than those without insurance. Patients who had four to seven ED visits or eight or more ED visits in one year were 2.63 and 3.10 times more likely to screen positive on the PRIME-MD, respectively, compared with those who had one to three visits. Patients who activated 911 two to ten times or 11 or more times in one year were 2.02 and 4.99 times more likely to screen positive for panic disorder, respectively, compared with those who had never activated 911. CONCLUSIONS: Patients who screen positive for panic disorder use emergency medical services and ED services more frequently. In addition, the overall prevalence of screening positive for panic disorder in an ED is higher than previously reported. PMID- 14525740 TI - The emergency severity index triage algorithm version 2 is reliable and valid. AB - OBJECTIVES: Initial studies have shown improved reliability and validity of a new triage tool, the Emergency Severity Index (ESI), over conventional three-level scales at two university medical centers. After pilot implementation and validation, the ESI was revised to include pediatric and updated vital signs criteria. The goal of this study was to assess ESI version (v.) 2 reliability and validity at seven emergency departments (EDs) in three states. METHODS: In part 1, interrater reliability was assessed using weighted kappa analysis of written training cases and postimplementation by a random sampling of actual patient triages. In part 2, validity was analyzed using a prospective cohort with stratified random sampling at each site. The ESI was compared with outcomes including resource consumption, inpatient admission, ED length of stay, and 60 day all-cause mortality. RESULTS: Weighted kappa analysis of interrater reliability ranged from 0.70 to 0.80 for the written scenarios (n = 3289) and 0.69 to 0.87 for patient triages (n = 386). Outcomes for the validity cohort (n = 1042) included hospitalization rates by ESI triage level: level 1, 83%; 2, 67%; 3, 42%; 4, 8%; level 5, 4%. Sixty-day all-cause mortality by triage level was as follows: level 1, 25%; 2, 4%; 3, 2%; 4, 1%; and 5, 0%. CONCLUSIONS: ESI v. 2 triage produced reliable, valid stratification of patients across seven sites. ESI triage should be evaluated as an ED casemix identification system for uniform data collection in the United States and compared with other major ED triage methods. PMID- 14525741 TI - How patients and visitors to an urban emergency department view clinical research. AB - OBJECTIVES: To assess views about clinical research, drawing current opinion from an urban, largely minority population within the authors' emergency department (ED). METHODS: Two focus groups of ED patients and visitors were conducted. These data informed the development of a 27-item interview examining views about clinical research and knowledge of human subjects protections. RESULTS: The authors interviewed a total of 172 patients and visitors within an adult ED. Study participants reflected the diverse patient population: 38% were African American, 32% Hispanic, 25% white, and 6% other. When asked why one might choose to participate in medical research, 46% said to benefit mankind, 26% said to improve one's own health, 18% cited access to medical care, 17% said financial incentive, and 11% said curiosity. When asked why one might decline research participation, 38% cited fear, 24% cited lack of interest in research, 10% cited medical mistrust, 9% indicated not wanting to feel like a "guinea pig," 6% indicated lack of time, and 5% suggested privacy concerns. When asked about the meaning of informed consent, 32% did not know. Many respondents (26%) were unaware that they could withdraw from a study. Although the majority (96%) endorsed a statement about the potential benefit of research for themselves or their loved ones, a sizable proportion of respondents (49%) equated research subjects to "human guinea pigs." CONCLUSIONS: Although many individuals tend to view clinical research favorably, a level of medical mistrust exists. The concerns about human experimentation and the limited understanding of human subject protections underscore the need to improve informed consent. PMID- 14525742 TI - Clinicopathological conference: massive hematemesis following blunt abdominal trauma. PMID- 14525743 TI - Emergency department overcrowding: what is our response to the "new normal"? PMID- 14525744 TI - Is spirituality an emergency physician's competency? PMID- 14525745 TI - National EMS research agenda: proceedings of the implementation symposium. AB - The National EMS Research Agenda identified eight recommendations for improving the conduct of emergency medical services (EMS) research in the United States. EMS leaders from across the country attended a two-day symposium to discuss implementation of the Research Agenda recommendations. The participants suggested specific methods to move the recommendations forward. These included improving training opportunities for EMS researchers, stimulating increases in available funding sources, facilitating the integration of research into practice, and crafting alterations within the regulatory environment. Participants felt that EMS must be more broadly integrated into the public health continuum. Federal agencies, states, local governments, charitable foundations, and corporations are asked to examine their practices to increase opportunities for participation in EMS research programs at all stages of the process. PMID- 14525746 TI - Strategies of disaster response in the health care system for tropical cyclones: experience following Typhoon Nari in Taipei City. AB - Natural disasters present significant potential for injuries and death. Unlike the experience of Hurricane Andrew that destroyed a vast surface area in the rural countryside, Typhoon Nari in Taipei proved that significant damages from natural disasters also can happen to modern health care systems in urban areas. To ameliorate such damages, specific structural, nonstructural, and administrative issues must be taken into account. Such issues include the location of the health facility, design of the infrastructure, storage of equipment and machines, maintenance, medical, and nonmedical operations. Specific considerations, such as early evacuation and securing the safety of the patients before a disaster, should be emphasized. Recovery plans that determine how soon medical service can be restored to the community should also be established. Emphasis on emergency-response preparedness, mitigation procedures, and recovery efforts should all be included in a comprehensive emergency plan against the destruction caused by natural hazards. PMID- 14525747 TI - Faculty development: academic opportunities for emergency medicine faculty on education career tracks. AB - Medical school faculty members who specialize in the scholarship of teaching have unique requirements for academic advancement in universities with clinician educator series. While excellence in teaching is the cornerstone of achievement, attention to traditional academic pursuits improves the likelihood of a favorable review by the institution's promotion and tenure committee. The teaching portfolio is an effective means to document performance. Ongoing faculty development and sound mentoring relationships facilitate the academic advancement of clinician-educators. PMID- 14525748 TI - Preventive care in the emergency department: screening for domestic violence in the emergency department. AB - OBJECTIVES: The most effective methods for identification and management of domestic violence (DV) victims in health care settings are unknown. The objective of this study was to systematically review screening for DV in the emergency department (ED) to identify victims and decrease morbidity and mortality from DV. METHODS: Using the terms "domestic violence" or "partner violence," and "identification" or "screening," and "emergency," the authors searched MEDLINE, the Cochrane Database, and Emergency Medical Abstracts from 1980-2002. They selected articles studying screening tools, interventions, or determining the incidence or prevalence of DV among ED patients. The studies were analyzed using evidence-based methodology. RESULTS: Three hundred thirty-nine articles resulted from the literature search. Based on selection criteria, 45 were reviewed in detail and 17 pertained to the ED. From references of these 17 articles, three additional articles were added. Screening can be conducted using a brief verbal screen and existing ED personnel. A randomized, controlled trial did not demonstrate a difference in screening rates between experimental and control hospitals. No studies assessed the effect of ED screening for DV on morbidity or mortality of domestic violence. An ED-based advocacy program resulted in increased use of shelters and counseling. CONCLUSIONS: Because of the paucity of outcomes research evaluating ED screening and interventions, there is insufficient evidence for or against DV screening in the ED. However, because of the high burden of suffering caused by DV, health care providers should strongly consider routinely inquiring about DV as part of the history, at a minimum for all female adolescent and adult patients. PMID- 14525750 TI - Usefulness of computerized pediatric motor vehicle safety discharge instructions. AB - OBJECTIVES: To determine whether providing child motor vehicle safety recommendations on computerized discharge instructions (CDIs) were useful to parents and modified their use of child-restraint devices (CRDs). METHODS: The subjects were guardians of children seen in an urban pediatric emergency department (ED). An intervention group was given computerized ED discharge instructions that included the National Highway Traffic Safety Administration recommendations for motor vehicle CRDs. A control group was given CDIs without the recommendations. All subjects were subsequently called within four days of the ED visit and asked questions about their knowledge and use of CRDs. They also were queried if the recommendations affected their knowledge or changed their behavior. RESULTS: There were 52 subjects in the control group and 58 in the intervention group. Fifty-seven percent of the intervention group remembered reading a safety tip (p < 0.001, 95% confidence interval [CI] = 0.32 to 0.62), and 82% of that subset correctly identified it pertaining to motor vehicle safety. Forty-five percent (n = 33) of those who remembered the safety recommendation in the intervention group supported it being educational (p = 0.067, 95% CI = 0.28 to 0.64). Ten percent of the subjects in the intervention group said the CDIs changed their behavior regarding buckling-up their child, compared with 0% of the control group (p = 0.473, 95% CI = 0.05 to 0.32). CONCLUSIONS: Including CRD information on CDIs is a convenient method of educating guardians of patients about motor vehicle safety in a pediatric ED setting. The data suggest that parents find it educational and a smaller subgroup change their behaviors after receiving them. PMID- 14525749 TI - Clinically important change in the visual analog scale after adequate pain control. AB - OBJECTIVES: To define the minimum clinically important difference (MCID) for the visual analog scale (VAS) of pain severity by measuring the change in VAS associated with adequate pain control. METHODS: The authors conducted a prospective, observational study. Adult emergency department (ED) patients with acute pain (<72 hours) were eligible. Patients rated their pain severity on a 100 mm VAS on presentation and at discharge. Patients were asked if they would accept any analgesic, then if they would accept a parenteral analgesic before treatment. At discharge, they were asked whether they had received adequate pain control. RESULTS: The authors enrolled 143 patients (mean age, 36 years; 54% female). The mean decrease in VAS was -30.0 mm (95% confidence interval [CI] = -36.4 to -23.6) for the 116 of 143 (81%) patients with adequate pain control at discharge vs. 5.7 (95% CI = -11.2 to -0.3) for the 27 with inadequate pain control (p < 0.001). At discharge, the mean VAS was 31.3 mm for those with adequate pain control vs. 55.1 for those without. Mean VAS for the 114 of 143 patients who would accept any analgesics initially was 64.7 vs. 47.1 for the 29 reporting no analgesic need. Initially, 77 patients would accept parenteral analgesics (mean VAS = 72.5 mm). CONCLUSIONS: A mean reduction in VAS of 30.0 mm represents a clinically important difference in pain severity that corresponds to patients' perception of adequate pain control. Defining MCID based on adequate analgesic control rather than minimal detectable change may be more appropriate for future analgesic trials, when effective treatments for acute pain exist. PMID- 14525751 TI - Evaluation of a new high-viscosity octylcyanoacrylate tissue adhesive for laceration repair: a randomized, clinical trial. AB - OBJECTIVE: Tissue adhesives have recently been approved for skin closure. Their low viscosity may result in inadvertent migration. The authors compared the tendency of the adhesive to migrate after laceration closure with a high- or low viscosity octylcyanoacrylate (OCA). METHODS: This was a randomized, clinical trial set in university and community-based emergency departments. Participants included patients with simple traumatic lacerations. Patients were randomized to laceration closure with low- or high-viscosity OCA tissue adhesive. The outcome measured was immediate adhesive migration (interobserver agreement, kappa = 0.90). Data analysis was performed with proportions compared with chi-square and Fisher's exact tests. RESULTS: Eighty-four patients were randomized to low- (n = 42) or high- (n = 42) viscosity OCA tissue adhesive. Groups were similar in baseline patient and wound characteristics. The high-viscosity OCA was less likely to migrate than the lower-viscosity agent (21% vs. 78%, p < 0.001; odds ratio = 0.3, 95% confidence interval = 0.1 to 0.5). The proportion of patients who noted a sensation of heat during OCA application was higher in the high viscosity groups (44% vs. 26% respectively, p = 0.11); however, all such patients in both groups would use the device again. At 14 days, there were no wound infections in either group. There was one dehiscence in the high-viscosity group. CONCLUSIONS: The high-viscosity OCA tissue adhesive was less likely to migrate than the lower-viscosity device. Wound dehiscence and infection rates were acceptably low in both treatment groups. PMID- 14525752 TI - Emergency medicine subinternship: can we provide a standard clinical experience? AB - Medical students have varied experiences on the emergency medicine (EM) subinternship. Didactic curricula can be standardized. OBJECTIVES: To determine if uniformity in clinical curricula is possible by assessing whether students can see patients with certain chief complaints (CC). METHODS: Prospective interventional analysis at a public teaching hospital. Control group (CG) students saw patients of their choice and recorded encounters in logbooks. Test group (TG) students were asked to see at least one patient with: orthopedic injury (OR); asthma exacerbation (AE); acute coronary syndrome (ACS); traumatic injury (TR); laceration (LAC); or diabetic ketoacidosis (DKA). TG students prospectively recorded these patients on a separate logbook page. Logbooks were reviewed by two investigators to determine if a patient with each diagnosis was seen. Chi-square analysis tested for differences in sample proportions between TG and CG. Multivariate analyses controlled for TG, specialty choice, and gender. p < 0.05 represented statistical significance. RESULTS: One hundred fifty (88 TG; 62 CG) students participated. Differences existed between TG and CG in the proportion of students who saw a patient with each CC: OR: 93% TG, 69% CG (p < 0.0001); AE: 86% TG, 63% CG (p < 0.0008); ACS: 97% TG, 58% CG (p < 0.0001); TR: 97% TG, 58% CG (p < 0.0001); LAC: 98% TG, 89% CG (p < 0.0220); DKA: 68% TG, 47% CG (p < 0.0086). Logistic regressions explaining the probability of seeing each CC showed the variable controlling for TG was positive and significant for 5 CCs: p = 0.0013 (OR); 0.0038 (AE); 0.0001 (ACS); 0.0001 (TR); 0.0229 (DKA). No difference was found for LAC: p = 0.0570. CONCLUSIONS: Students can be directed to see patients with particular CCs. TG students saw more patients with certain CCs than CG students, p < 0.0001. This intervention can help educators provide a well-rounded, uniform clinical EM experience. PMID- 14525754 TI - Organophosphate poisoning. PMID- 14525756 TI - Emergency care transitions. PMID- 14525757 TI - Ultrasound training. PMID- 14525759 TI - The effect of human tissue factor pathway inhibitor-2 on the growth and metastasis of fibrosarcoma tumors in athymic mice. AB - Human tissue factor pathway inhibitor-2 (TFPI-2) is a matrix-associated Kunitz inhibitor that inhibits the plasmin- and trypsin-mediated activation of zymogen matrix metalloproteinases involved in tumor progression, invasion, and metastasis. To directly assess its role in tumor growth and metastasis in vivo, we stably transfected HT-1080 fibrosarcoma cells expressing either fully active wild-type human TFPI-2 (WT) or inactive R24Q TFPI-2 (QT) and examined their ability to form tumors and metastasize in athymic mice in comparison to mock transfected cells (MT). MT and QT fibrosarcoma tumors grew 2 to 3 times larger than WT tumors. Tumor metastasis was confined to the lung and was observed in 75% of mice treated with either MT or QT cells, whereas only 42% of mice treated with WT cells developed lung metastases. Real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR) analyses of each tumor group revealed 3- to 6 fold lower levels of murine vascular endothelial growth factor gene expression in WT tumors in relation to either MT or QT tumors. Comparative tumor gene expression analysis revealed that several human genes implicated in oncogenesis, invasion, metastasis, apoptosis, and angiogenesis had significantly altered levels of expression in WT tumors. Our collective data demonstrate that secretion of inhibitory TFPI-2 by a highly metastatic tumor cell markedly inhibits its growth and metastasis in vivo by regulating pericellular extracellular matrix (ECM) remodeling and angiogenesis. PMID- 14525760 TI - Bifunctional role for VEGF-induced heme oxygenase-1 in vivo: induction of angiogenesis and inhibition of leukocytic infiltration. AB - Heme-oxygenases (HOs) catalyze the conversion of heme into carbon monoxide and biliverdin. HO-1 is induced during hypoxia, ischemia/reperfusion, and inflammation, providing cytoprotection and inhibiting leukocyte migration to inflammatory sites. Although in vitro studies have suggested an additional role for HO-1 in angiogenesis, the relevance of this in vivo remains unknown. We investigated the involvement of HO-1 in angiogenesis in vitro and in vivo. Vascular endothelial growth factor (VEGF) induced prolonged HO-1 expression and activity in human endothelial cells and HO-1 inhibition abrogated VEGF-driven angiogenesis. Two murine models of angiogenesis were used: (1) angiogenesis initiated by addition of VEGF to Matrigel and (2) a lipopolysaccharide (LPS) induced model of inflammatory angiogenesis in which angiogenesis is secondary to leukocyte invasion. Pharmacologic inhibition of HO-1 induced marked leukocytic infiltration that enhanced VEGF-induced angiogenesis. However, in the presence of an anti-CD18 monoclonal antibody (mAb) to block leukocyte migration, VEGF-induced angiogenesis was significantly inhibited by HO-1 antagonists. Furthermore, in the LPS-induced model of inflammatory angiogenesis, induction of HO-1 with cobalt protoporphyrin significantly inhibited leukocyte invasion into LPS-conditioned Matrigel and thus prevented the subsequent angiogenesis. We therefore propose that during chronic inflammation HO-1 has 2 roles: first, an anti-inflammatory action inhibiting leukocyte infiltration; and second, promotion of VEGF-driven noninflammatory angiogenesis that facilitates tissue repair. PMID- 14525761 TI - Treatment of CD40 ligand deficiency by hematopoietic stem cell transplantation: a survey of the European experience, 1993-2002. AB - CD40 ligand (CD40L) deficiency causes recurrent sinopulmonary infection, Pneumocystis carinii pneumonia, and Cryptosporidium parvum infection. Approximately 40% to 50% of patients survive to the third decade: long-term survival is unclear. Hematopoietic stem cell transplantation (HSCT) is curative. We present a retrospective analysis of 38 European patients undergoing HSCT for CD40L deficiency in 8 European countries between 1993 and 2002. Donor stem cell source included 14 HLA-identical siblings, 22 unrelated donors, and 2 phenotypically matched parental stem cells (12 T-cell depleted). Of the patients, 34 engrafted and 26 (68%) survived; 3 had autologous reconstitution, 22 (58%) were cured, and 1 engrafted but has poor T-cell immune reconstitution. There were 18 evaluated patients who responded to vaccination. Of the patients, 12 (32%) died from infection-related complications, with severe cryptosporidiosis in 6. Grades 2 to 4 graft-versus-host disease (GvHD) associated with infection occurred in 6 of 12 fatal cases. HSCT cured 58% of patients, 72% of those without hepatic disease. Early T-cell function following whole marrow HSCT may limit cryptosporidial disease, but survival was similar after T-cell-depleted HSCT. Preexisting lung damage was the most important adverse risk factor. Further studies will determine optimal timing and type of HSCT. PMID- 14525762 TI - Haploinsufficiency of Runx1 results in the acceleration of mesodermal development and hemangioblast specification upon in vitro differentiation of ES cells. AB - The AML1 gene (recently renamed Runx1), which encodes the DNA-binding subunit of a transcription factor of the core binding factor (CBF) family, is required for the establishment of definitive hematopoiesis. We have previously demonstrated that Runx1 is expressed in yolk sac mesodermal cells prior to the establishment of the blood islands and in the embryoid body (EB)-derived blast-colony-forming cells (BL-CFCs), the in vitro equivalent of the hemangioblast. Analysis of Runx1 deficient embryonic stem (ES) cells demonstrated that this gene is essential for the generation of normal numbers of blast colonies, the progeny of the BL-CFCs. In the present study, we analyzed the potential of Runx1(+/-) ES cells to determine if heterozygosity at the Runx1 locus impacts early developmental events leading to the commitment of the BL-CFCs. Our results indicate that Runx1 heterozygosity leads to an acceleration of mesodermal commitment and specification to the BL-CFCs and to the hematopoietic lineages in EBs. PMID- 14525763 TI - Vascular endothelial growth factor (VEGF) induces rapid prourokinase (pro-uPA) activation on the surface of endothelial cells. AB - Vascular endothelial growth factor (VEGF) is the pivotal angiogenic growth factor activating endothelial cells to migrate, proliferate, and form capillary tubes. For an ordered endothelial cell migration, tissue invasion, and degradation of the extracellular matrix, proteolytic machinery is indispensable. Such machinery, suitable for localized proteolysis, is provided by the prourokinase-urokinase plasmin system. Prourokinase (pro-uPA), the initial component of this system, is, however, synthesized in its inactive precursor form and as such bound to its cellular receptor uPAR. Here we identify a mechanism via which VEGF(165) interacting with its receptor VEGFR-2 rapidly induces prourokinase activation that is dependent on a change in integrin affinity, activation of matrix metalloproteinase 2 (MMP-2), and pro-uPA being bound to its surface receptor uPAR. This VEGF-induced pro-uPA activation on endothelial cells is responsible for VEGF-dependent local fibrinolytic activity and might be one of the initial steps in the angiogenic process. PMID- 14525764 TI - Association of CBFA2 mutation with decreased platelet PKC-theta and impaired receptor-mediated activation of GPIIb-IIIa and pleckstrin phosphorylation: proteins regulated by CBFA2 play a role in GPIIb-IIIa activation. AB - The mechanisms by which agonists activate glycoprotein (GP) IIb-IIIa function remain unclear. We have reported data on a patient with thrombocytopenia and impaired receptor-mediated aggregation, phosphorylation of pleckstrin (a protein kinase C [PKC] substrate), and activation of the GPIIb-IIIa complex. Abnormalities in hematopoietic transcription factors have been associated with thrombocytopenia and platelet dysfunction. To define the molecular mechanisms, we amplified from patient platelet RNA exons 3 to 6 of core-binding factor A2 (CBFA2) cDNA, which encompasses the DNA-binding Runt domain; a 13-nucleotide (nt) deletion was found (796-808 nt). The gDNA revealed a heterozygous mutation (G>T) in intron 3 at the splice acceptor site for exon 4, leading to a frameshift with premature termination in the Runt domain. On immunoblotting, platelet CBFA2, PKC , albumin, and IgG were decreased, but pleckstrin, PKC-alpha, -betaI, -betaII, eta, -epsilon, -delta, and -zeta, and fibrinogen were normal. Our conclusions are that (1) CBFA2 mutation is associated with not only thrombocytopenia, but also impaired platelet protein phosphorylation and GPIIb-IIIa activation; (2) proteins regulated by CBFA2 are required for inside-out signal transduction-dependent activation of GPIIb-IIIa; and (3) we have documented the first deficiency of a human PKC isozyme (PKC-), suggesting a major role of this isozyme in platelet production and function. PMID- 14525765 TI - Activated Fps/Fes partially rescues the in vivo developmental potential of Flk1 deficient vascular progenitor cells. AB - Relatively little is known about the modulators of the vascular endothelial growth factor A (VEGF-A)/Flk1 signaling cascade. To functionally characterize this pathway, VEGF-A stimulation of endothelial cells was performed. VEGF-A mediated Flk1 activation resulted in increased translocation of the endogenous Fps/Fes cytoplasmic tyrosine kinase to the plasma membrane and increased tyrosine phosphorylation, suggesting a role for Fps/Fes in VEGF-A/Flk1 signaling events. Addition of a myristoylation consensus sequence to Fps/Fes resulted in VEGF-A independent membrane localization of Fps/Fes in endothelial cells. Expression of the activated Fps/Fes protein in Flk1-deficient embryonic stem (ES) cells rescued their contribution to the developing vascular endothelium in vivo by using ES cell-derived chimeras. Activated Fps/Fes contributed to this rescue event by restoring the migratory potential to Flk1 null progenitors, which is required for movement of hemangioblasts from the primitive streak region into the yolk sac proper. Activated Fps/Fes in the presence of Flk1 increased the number of hemangioblast colonies in vitro and increased the number of mesodermal progenitors in vivo. These results suggest that Fps/Fes may act synergistically with Flk1 to modulate hemangioblast differentiation into the endothelium. We have also demonstrated that activated Fps/Fes causes hemangioma formation in vivo, independently of Flk1, as a result of increasing vascular progenitor density. PMID- 14525766 TI - Adoptive immunotherapy with donor lymphocyte infusions after allogeneic hematopoietic cell transplantation following nonmyeloablative conditioning. AB - This study retrospectively analyzed data from 446 patients given hematopoietic cell transplants from HLA-matched related or unrelated donors after conditioning with 2 Gy total body irradiation with or without fludarabine and postgrafting immunosuppression with mycophenolate mofetil and cyclosporine following grafting. Fifty-three of 446 patients received donor lymphocyte infusion (DLI) with a median CD3 dose of 1 x 10(7) cells/kg. Their diagnoses included myelodysplastic syndrome (n = 10), acute leukemia (n = 10), chronic leukemia (n = 11), multiple myeloma (n = 9), lymphoma (n = 9), and solid tumors (n = 4). Patients received DLI for persistent disease (n = 8), disease relapse (n = 17), progressive disease (n = 12), low donor chimerism with disease (n = 11), or low chimerism with disease remission (n = 5). Seventeen of the 53 patients (32%) are alive with a median follow-up of 30 months; 5 are in complete remission (CR), 2 are in partial remission (PR), and 10 have stable or progressive disease. Nine of 53 patients (17%) developed grades II to IV acute graft-versus-host disease. Of 48 patients receiving DLI for treatment of disease, 7 achieved CR and 5 PR, with an overall response rate of 25%. Six of 16 patients who received DLI for chimerism had increases in donor chimerism leading to sustained engraftment, whereas 10 eventually rejected their grafts. In conclusion, DLI is a potential treatment strategy, with acceptable toxicity, for patients with persistent, relapsed, or progressive disease after nonmyeloablative hematopoietic cell transplantation. PMID- 14525767 TI - Hypoxic gene activation by lipopolysaccharide in macrophages: implication of hypoxia-inducible factor 1alpha. AB - Hypoxia-inducible factor 1 (HIF-1) regulates many genes induced by low oxygen conditions. The expression of important hypoxic genes such as glucose transporter 1 and vascular endothelial growth factor are increased in macrophages during wound healing and in the presence of the endotoxin, lipopolysaccharide (LPS). Recent studies have demonstrated that nonhypoxic stimuli can also activate HIF-1 in a cell-specific manner. Here, we demonstrate that in macrophages, LPS can control the activation of hypoxia-regulated genes through the HIF-1 pathway. We show that in these cells, protein expression levels of HIF-1alpha are strongly increased to levels comparable to hypoxic induction. HIF-1alpha mRNA levels are markedly increased following LPS stimulation, suggesting a transcriptional induction. In functional studies, the LPS-induced HIF-1 complex could specifically bind to the HIF-1 DNA-binding motif. Additionally, when cells were transfected with an HIF-1-specific reporter construct, LPS could strongly activate the expression of the reporter to levels that surpassed those observed after hypoxic induction. This induction was blocked by the cotransfection of a dominant-negative form of HIF-1alpha. These results indicate that the HIF-1 complex is involved in macrophage gene activation following LPS exposure and identify a novel pathway that could play a determinant role during inflammation and wound healing. PMID- 14525768 TI - Fiber-modified adenoviruses generate subgroup cross-reactive, adenovirus-specific cytotoxic T lymphocytes for therapeutic applications. AB - Adenovirus (Ad) infections are responsible for considerable morbidity and mortality, particularly in pediatric hematopoietic stem cell transplant (HSCT) recipients. To date there is no therapy. The present study was motivated by the potential for using adoptive immunotherapy as either prophylaxis or treatment for Ad infections and associated diseases. The authors have developed a protocol to reactivate Ad-specific memory T cells from peripheral blood mononuclear cells (PBMCs) using a clinical-grade adenoviral vector. Such lines contain a specific CD4 and CD8 T-cell component and are capable of recognizing and lysing target cells infected with wild-type Ad serotypes from different Ad groups. Furthermore, the frequency of Ad-specific precursors can be determined in PBMCs ex vivo and used as a means to assess changes in Ad-specific T-cell memory responses after infusion. This is the first report of a simple and reproducible method to activate and expand Ad-specific cytotoxic T lymphocytes (CTLs), which should be protective against the range of different Ad subtypes that affect transplant recipients. PMID- 14525769 TI - The MHC class II transcriptional activator (CIITA) inhibits HTLV-2 viral replication by blocking the function of the viral transactivator Tax-2. AB - The human T-cell leukemia virus type 2 (HTLV-2), an oncogenic retrovirus closely related to HTLV-1, produces a lifelong infection whose possible association to certain human diseases is still debated. Although some viral products can influence the expression and action of cellular genes, very little is known about the molecular mechanisms involved. Here we show that the AIR-1-encoded human major histocompatibility complex (MHC) class II transactivator (CIITA) strongly inhibits viral replication, but not virus entry, in human B- and T-cell susceptible targets. This effect results from CIITA inhibiting the Tax-mediated transactivation of the HTLV-2 long-term repeat. Further molecular analysis shows that the N-terminal region of CIITA encompassing the first 321 amino acids is responsible for the inhibitory effect on viral replication. This region is crucial for the transactivation of human MHC class II genes and includes the activation domain as well as domains interacting with coactivators that also are used by the viral transactivator Tax to modulate cellular functions. These results represent the first evidence that a cellular transcriptional activator, controlling the coordinate expression of the entire family of MHC class II antigen-presenting molecules, inhibits HTLV-2 viral replication by a distinct mechanism. In this new role CIITA may represent a new tool for therapeutic strategies aimed at counteracting HTLV-2 replication and spreading. PMID- 14525770 TI - Itraconazole versus fluconazole for prevention of fungal infections in patients receiving allogeneic stem cell transplants. AB - Prophylactic fluconazole prevents candidiasis; however, this drug has no activity against molds. We performed a randomized trial to determine whether prophylactic itraconazole prevents invasive mold infections (IMIs). A total of 304 patients receiving allogeneic stem cell transplants (SCT) were randomized to receive fluconazole (400 mg/d) or itraconazole (oral solution 2.5 mg/kg 3 times daily, or intravenous 200 mg daily) for 180 days after SC transplantation, or until 4 weeks after discontinuation of graft-versus-host disease (GVHD) therapy. Proven or probable invasive fungal infections (IFI) were evaluated by intent-to-treat and "on-treatment" analyses. More patients in the itraconazole arm developed hepatotoxicities, and more patients were discontinued from itraconazole because of toxicities or gastrointestinal (GI) intolerance (36% versus 16%, P <.001). Intent-to-treat analysis demonstrated no difference in the incidence of IFI during the intended study period (fluconazole 16% versus itraconazole 13%, P =.46); however, fewer patients in the itraconazole arm developed IFI on treatment (fluconazole 15% versus itraconazole 7%, P =.03). Itraconazole provided better protection against IMI (fluconazole 12% versus itraconazole 5%, P =.03), but similar protection against candidiasis (3% versus 2%, P =.69). There was no difference in overall or fungal-free survival. Itraconazole appears to prevent IMI in the subset of patients who tolerate the drug; however, toxicities and poor tolerability limit its success as prophylactic therapy. PMID- 14525771 TI - Association of CD26 with CD45RA outside lipid rafts attenuates cord blood T-cell activation. AB - CD26 is a T-cell activation antigen that contains dipeptidyl peptidase IV activity and binds adenosine deaminase. Recent work showed that specialized membrane microdomains, also known as lipid rafts, play a key role in T-cell signaling. In this study, we investigate the role of CD26 in cord blood T-cell activation and signal transduction. We demonstrated that different expression levels of CD26 were observed between cord blood T cells (CBTCs) and peripheral blood T cells (PBTCs) and that CD26(+)CD45RA(+) CBTCs were different compared with CD26(+)CD45RA(+) PBTCs. Moreover, the comitogenic effect of CD26 was not as pronounced in CBTCs as in PBTCs. We also showed that CD26 cross-linking induced less phosphorylation of T-cell receptor-signaling molecules, lymphoid T-cell protein tyrosine kinase (Lck), zeta-associated protein 70 (ZAP-70), T-cell receptor zeta (TCRzeta), and linker for activator of T cells (LAT) in CBTCs than in PBTCs. Furthermore, CD26 molecules associated with CD45RA molecules outside lipid rafts in CBTCs. Our results suggest that strong physical linkage of CD26 with CD45RA outside lipid rafts may be responsible for the attenuation of T-cell activation signaling through CD26, which may be responsible for immature immune response and the low incidence of severe graft-versus-host disease in cord blood transplantation. PMID- 14525772 TI - Incomplete restoration of colony-stimulating factor 1 (CSF-1) function in CSF-1 deficient Csf1op/Csf1op mice by transgenic expression of cell surface CSF-1. AB - The primary macrophage growth factor, colony-stimulating factor 1 (CSF-1), is expressed as a secreted glycoprotein or proteoglycan found in the circulation or as a biologically active cell surface glycoprotein (csCSF-1). To investigate the in vivo roles of csCSF-1, we created mice that exclusively express csCSF-1, in a normal tissue-specific and developmental manner, by transgenic expression of csCSF-1 in the CSF-1-deficient osteopetrotic (Csf1(op)/Csf1(op)) background. The gross defects of Csf1(op)/Csf1(op) mice, including growth retardation, failure of tooth eruption, and abnormal male and female reproductive functions were corrected. Macrophage densities in perinatal liver, bladder, sublinguinal salivary gland, kidney cortex, dermis, and synovial membrane were completely restored, whereas only partial or no restoration was achieved in adult liver, adrenal gland, kidney medulla, spleen, peritoneal cavity, and intestine. Residual osteopetrosis, significantly delayed trabecular bone resorption in the subepiphyseal region of the long bone, and incomplete correction of the hematologic abnormalities in the peripheral blood, bone marrow, and spleens of CSF-1-deficient mice were also found in mice exclusively expressing csCSF-1. These data suggest that although csCSF-1 alone is able to normalize several aspects of development in Csf1(op)/Csf1(op) mice, it cannot fully restore in vivo CSF-1 function, which requires the presence of the secreted glycoprotein and/or proteoglycan forms. PMID- 14525773 TI - Gene microarray analysis reveals interleukin-5-dependent transcriptional targets in mouse bone marrow. AB - Interleukin-5 (IL-5) is a hematopoietic differentiation factor that promotes the development of mature eosinophils from progenitors in bone marrow. We present a multifactorial microarray study documenting the transcriptional events in bone marrow of wild-type and IL-5-deficient mice at baseline and in response to infection with Schistosoma mansoni. The microarray data were analyzed by a 4-way subtractive algorithm that eliminated confounding non-IL-5-related sequelae of schistosome infection as well as alterations in gene expression among uninfected mice. Among the most prominent findings, we observed 7- to 40-fold increased expression of transcripts encoding the classic eosinophil granule proteins (eosinophil peroxidase, major basic protein, the ribonucleases) together with arachidonate-15-lipoxygenase and protease inhibitor plasminogen activator inhibitor 2 (PAI-2), in the IL-5-producing, infected wild-type mice only. This was accompanied by increased transcription of genes involved in secretory protein biosynthesis and granule-vesicle formation. Interestingly, we did not detect increased expression of genes encoding eosinophil-related chemokine receptors (CCR1, CCR3) or members of the GATA or CCAAT/enhancer binding protein (C/EBP) transcription factor families. These data suggest that the IL-5-responsive progenitors in the mouse bone marrow are already significantly committed to the eosinophil lineage and that IL-5 promotes differentiation of these committed progenitors into cells with recognizable and characteristic cytoplasmic granules and granule proteins. PMID- 14525774 TI - T-cell homeostasis in humans with thymic hypoplasia due to chromosome 22q11.2 deletion syndrome. AB - Patients with chromosome 22q11.2 deletion syndrome (DiGeorge syndrome/velocardiofacial syndrome) typically exhibit thymic hypoplasia, conotruncal cardiac defects, and hypoparathyroidism. The immunodeficiency that results from the thymic hypoplasia has been extensively described and consists primarily of T-cell lymphopenia. A curious feature of the T-cell lymphopenia is that the age-related rate of decline of T-cell numbers is slower in patients than controls. This leads to T-cell numbers in adulthood that are minimally decreased compared with controls. This suggests that homeostatic mechanisms might be acting to preserve the peripheral blood T-cell numbers in patients. We characterized changes in CD4/CD45RA and CD4/CD45RO T-cell populations in patients and controls of various ages and determined T-cell recombination excision circles and telomere length within the CD4/CD45RA population. Patients had evidence of accelerated conversion of naive to memory cells and had evidence of more extensive replicative history within the CD4/CD45RA compartment compared with controls. Oligoclonal T-cell receptor (TCR) Vbeta families and missing Vbeta families were seen more often in patients than controls. These data are consistent with homeostatic proliferation of T cells in patients with limited T-cell production due to thymic hypoplasia. PMID- 14525775 TI - Differential processing of stromal-derived factor-1alpha and stromal-derived factor-1beta explains functional diversity. AB - The chemokine stromal-derived factor-1 (SDF-1), which is constitutively expressed in most tissues as SDF-1alpha and SDF-1beta resulting from alternative gene splicing, regulates hematopoiesis, lymphocyte homing, B-lineage cell growth, and angiogenesis. Because SDF-1alpha and SDF-1beta are constitutively and ubiquitously expressed, their degradation must serve an important regulatory role. Here we show that SDF-1alpha and SDF-1beta are secreted as full-length molecules. When exposed to human serum, full-length SDF-1alpha (1-68) undergoes processing first at the COOH terminus to produce SDF-1alpha 1-67 and then at the NH2 terminus to produce SDF-1alpha 3-67. By contrast, full-length SDF-1beta (1 72) is processed only at the NH2 terminus to produce SDF-1beta 3-72. CD26/dipeptidyl peptidase is responsible for serum cleavage of SDF-1alpha and SDF 1beta at the NH2 terminus. Serum processing of SDF-1alpha at the COOH terminus, which has not been previously reported, reduces the ability of the polypeptide to bind to heparin and to cells and to stimulate B-cell proliferation and chemotaxis. The additional processing at the NH2 terminus renders both forms of SDF-1 unable to bind to heparin and to activate cells. The differential processing of SDF-1alpha and SDF-1beta provides biologic significance to the existence of 2 splice forms of the chemokine and adds a tool to precisely regulate SDF-1's biologic activity by changes in specific activity. PMID- 14525776 TI - Gene expression patterns associated with recurrent chromosomal translocations in acute lymphoblastic leukemia. AB - We obtained a global view of gene expression in both cell lines and pediatric acute lymphoblastic leukemia (ALL) samples that harbor one of several selected chromosomal abnormalities. When the cell lines were studied alone, we found that these chromosomal abnormalities were associated with the predominant variation in transcriptional programs across the set of cell lines studied. When cell lines and clinical samples were studied together, we found that each chromosomal abnormality (TEL/AML1, BCR/ABL, or MLL abnormalities) was associated with a characteristic gene expression signature that was shared by both cell lines and clinical samples. However, BCR/ABL was associated with a much more heterogeneous pattern of expression than were TEL/AML1 and MLL abnormalities. This observation has important implications for the study of BCR/ABL ALL. In addition, we systematically identified genes whose expression was associated with TEL/AML1, BCR/ABL, or MLL abnormalities in both clinical samples and cell lines. Although some of these genes have previously been described, many have not previously been reported to be associated with one of these chromosomal abnormalities. Notably, we found that the erythropoietin receptor (EPOR) is consistently highly expressed in TEL/AML1 ALL compared with BCR/ABL or MLL. PMID- 14525777 TI - Epidemiology of human parvovirus B19 in children with sickle cell disease. AB - Human parvovirus (HPV) B19 causes significant morbidity and mortality in children with sickle cell disease (SCD), but little data are published about the epidemiology of HPV B19 infection and its associated complications in this patient population. In this study, prevalence and incidence rates of HPV B19 were determined in 633 patients with SCD followed at The Children's Hospital of Philadelphia between November 1996 and December 2001. Thirty percent (30%) were HPV B19 immunoglobulin G (IgG) positive at first testing, and the 70% without evidence of past HPV B19 infection were tested annually. One hundred ten patients developed evidence of HPV B19 infection for an incidence rate of 11.3 per 100 patient years. Sixty-eight episodes of HPV B19-induced transient red cell aplasia occurred with the following clinical events: fever (89.7%), pain (61.8%), acute splenic sequestration (19.1%), and acute chest syndrome (11.8%). Pain, fever, and acute splenic sequestration were more frequent events with acute HPV B19 infections compared with acute events in uninfected patients. The results of this epidemiologic study, the largest and most comprehensive to date, justify the development of HPV B19 prevention strategies to diminish the frequent and often severe complications associated with HPV B19 infections in patients with SCD. PMID- 14525778 TI - Lymphodepletion in the ApcMin/+ mouse model of intestinal tumorigenesis. AB - Germ line mutations in the Adenomatous polyposis coli tumor suppressor gene cause a hereditary form of intestinal tumorigenesis in both mice and man. Here we show that in Apc(Min/+) mice, which carry a heterozygous germ line mutation at codon 850 of Apc, there is progressive loss of immature and mature thymocytes from approximately 80 days of age with complete regression of the thymus by 120 days. In addition, Apc(Min/+) mice show parallel depletion of splenic natural killer (NK) cells, immature B cells, and B progenitor cells in bone marrow due to complete loss of interleukin 7 (IL-7)-dependent B-cell progenitors. Using bone marrow transplantation experiments into wild-type recipients, we have shown that the capacity of transplanted Apc(Min/+) bone marrow cells for T- and B-cell development appears normal. In contrast, although the Apc(Min/+) bone marrow microenvironment supported short-term reconstitution with wild-type bone marrow, Apc(Min/+) animals that received transplants subsequently underwent lymphodepletion. Fibroblast colony-forming unit (CFU-F) colony assays revealed a significant reduction in colony-forming mesenchymal progenitor cells in the bone marrow of Apc(Min/+) mice compared with wild-type animals prior to the onset of lymphodepletion. This suggests that an altered bone marrow microenvironment may account for the selective lymphocyte depletion observed in this model of familial adenomatous polyposis. PMID- 14525779 TI - Assessment of removal of human cytomegalovirus from blood components by leukocyte depletion filters using real-time quantitative PCR. AB - To assess removal of cytomegalovirus (CMV) by leukocyte depletion (LD) filters, we developed a spiking model of latent virus using peripheral blood mononuclear cells (PBMCs) infected by coculture with CMV-infected human fibroblasts. Infected PBMCs were purified by dual magnetic column selection and then spiked into whole blood units or buffy coat pools prior to LD by filtration. CMV load and fibroblast contamination were assessed using quantitative CMV DNA real-time PCR and quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) of mRNA encoding the fibroblast-specific splice variant of prolyl-4-hydroxylase, respectively. After correcting for fibroblast-associated CMV, the mean CMV load was reduced in whole blood by LD from 7.42 x 10(2) to 1.13 copies per microliter (2.81(10)log reduction) and from 3.8 x 10(2) to 4.77 copies per microliter (1.9(10)log reduction) in platelets. These results suggest that LD by filtration reduces viral burden but does not completely remove CMV from blood components. PMID- 14525780 TI - Elevated interleukin-7 levels not sufficient to maintain T-cell homeostasis during simian immunodeficiency virus-induced disease progression. AB - Elevated levels of interleukin 7 (IL-7) have been correlated with various T-cell depletion conditions, including HIV infection, and suggested as an indicator of HIV disease progression (AIDS and death). Here, the assessment of pathogenic simian immunodeficiency virus (SIVmac239) infection in rhesus macaques demonstrated a clear association between a significant elevation in IL-7 levels and disease progression. In 5 macaques that progressed to simian AIDS and death, elevated IL-7 levels were unable to restore T-cell homeostasis. In contrast, increased IL-7 levels were followed by relatively high and stable T-cell numbers in the SIV-infected macaques with a slow-progressing phenotype. Further, studies in sooty mangabeys that do not progress to simian AIDS and that maintain stable T cell numbers despite high levels of viral replication support the importance of IL-7 and T-cell homeostasis in disease progression. These data suggest that during pathogenic SIV infection with high viral replication, elevated IL-7 levels are unable to recover T-cell homeostasis, thereby leading to disease progression. The utility of IL-7 as a potential immunotherapeutic agent to improve HIV/SIV related T-cell depletion may therefore depend on controlling the pathogenic effects of viral replication prior to the administration of IL-7. PMID- 14525781 TI - IFN-alpha promotes the rapid differentiation of monocytes from patients with chronic myeloid leukemia into activated dendritic cells tuned to undergo full maturation after LPS treatment. AB - Chronic myelogenous leukemia (CML) is a malignant myeloproliferative disease arising from the clonal expansion of a stem cell expressing the bcr/abl oncogene. CML patients frequently respond to treatment with interferon-alpha (IFN-alpha), even though the mechanisms of the response remain unclear. In the present study, we evaluated the role of IFN-alpha in differentiation and activity of monocyte derived dendritic cells (DCs) from CML patients as well as in modulation of the cell response to lipopolysaccharide (LPS). Treatment of CML monocytes with IFN alpha and granulocyte-macrophage colony-stimulating factor (GM-CSF) resulted in the rapid generation of activated DCs (CML-IFN-DCs) expressing interleukin-15 (IL 15) and the antiapoptotic bcl-2 gene. These cells were fully competent to induce IFN-gamma production by cocultured autologous T lymphocytes and expansion of CD8(+) T cells. LPS treatment of CML-IFN-DCs, but not of immature DCs generated in the presence of IL-4/GM-CSF, induced the generation of CD8(+) T cells reactive against autologous leukemic CD34(+) cells. Altogether, these results suggest that (1) the generation of highly active monocyte-derived DCs could be important for the induction of an antitumor response in IFN-treated CML patients and (2) IFN alpha can represent a valuable cytokine for the rapid generation of active monocyte-derived DCs to be utilized for vaccination strategies of CML patients. PMID- 14525782 TI - The in vitro production and characterization of neutrophils from embryonic stem cells. AB - An embryonic stem (ES) cell/OP9 coculture system for the effective production of functional neutrophils is described. A 3-step differentiation strategy was developed that uses liquid culture, enabling reliable and abundant production of neutrophils at high purity without the need of sorting for isolation of mature neutrophils. Use of the OP9 stromal cell line significantly enhances the number, percentage, and duration of differentiated neutrophils produced from embryonic stem cells. Effective and sustained differentiation of ES cells into neutrophils provides a useful model system for studying neutrophil differentiation and function and the factors that regulate them. Morphologic and functional evaluation of these ES-derived neutrophils indicates that large numbers of mature neutrophils can be produced from pluripotent ES cells in vitro. Specifically, their morphology, ability to produce superoxides, flux calcium, undergo chemotaxis in response to macrophage inflammatory protein 2 (MIP-2), stain for the granulocyte-specific marker-specific chloroacetate esterase, and contain the neutrophil-specific markers Gr-1 and the mouse neutrophil-specific antigen indicates that they are comparable with purified mouse bone marrow neutrophils. They also express gelatinase and lactoferrin granule proteins. During the differentiation of these ES-derived neutrophils, regional areas of neutrophil production can be identified that have been designated as neutrophil generating regions (NGRs). PMID- 14525783 TI - Ex vivo depletion of alloreactive cells based on CFSE dye dilution, activation antigen selection, and dendritic cell stimulation. AB - Eliminating alloreactive cells from T-cell populations would enable the transfer of immune function to patients who receive stem cell transplants. However, high efficiency depletion has proved difficult to achieve. We sought to develop ex vivo approaches for the maximal depletion of alloreactive CD4(+) T cells. Using a flow cytometric cell sorting approach after mixed lymphocyte reaction (MLR) culture, we have found that sorted CFSE(bright) (5-(and-6)-carboxyfluorescein diacetate succinmidyl ester) (nondivided) and activation antigen-negative cells are markedly depleted of alloreactivity. With HLA-mismatched peripheral blood mononuclear cell (PBMC) stimulators we have consistently attained (90%-95%) depletion of alloreactivity. Importantly, when purified matured monocyte-derived dendritic cells (DCs) are used as stimulators, a 100-fold (99%) reduction in alloreactivity was attained, resulting in abrogation of the secondary MLR. Significantly, the CFSE(bright) CD25(-) cells recovered from these cultures retained general immunoreactivity, including responses to Candida and cytomegalovirus (CMV) antigens. In addition, a CFSE-based approach was tested and found to be sufficient for graft-versus-host disease (GVHD) prevention in vivo, in a major histocompatibility complex (MHC) class II disparate murine model. This efficient approach to selectively deplete mature alloantigen-specific T cells may permit enhanced immune reconstitution without GVHD. PMID- 14525784 TI - Development of both human connective tissue-type and mucosal-type mast cells in mice from hematopoietic stem cells with identical distribution pattern to human body. AB - The transplantation of primitive human cells into sublethally irradiated immune deficient mice is the well-established in vivo system for the investigation of human hematopoietic stem cell function. Although mast cells are the progeny of hematopoietic stem cells, human mast cell development in mice that underwent human hematopoietic stem cell transplantation has not been reported. Here we report on human mast cell development after xenotransplantation of human hematopoietic stem cells into nonobese diabetic severe combined immunodeficient (NOD/SCID)/gamma(c)(null) (NOG) mice with severe combined immunodeficiency and interleukin 2 (IL-2) receptor gamma-chain allelic mutation. Supported by the murine environment, human mast cell clusters developed in mouse dermis, but they required more time than other forms of human cell reconstitution. In lung and gastric tract, mucosal-type mast cells containing tryptase but lacking chymase located on gastric mucosa and in alveoli, whereas connective tissue-type mast cells containing both tryptase and chymase located on gastric submucosa and around major airways, as in the human body. Mast cell development was also observed in lymph nodes, spleen, and peritoneal cavity but not in the peripheral blood. Xenotransplantation of human hematopoietic stem cells into NOG mice can be expected to result in a highly effective model for the investigation of human mast cell development and function in vivo. PMID- 14525785 TI - Donor immune reconstitution after liver-small bowel transplantation for multiple intestinal atresia with immunodeficiency. AB - The syndrome of multiple intestinal atresia with immunodeficiency is a rare, invariably fatal congenital disorder. At 16 months of age, a child with this syndrome underwent liver-small bowel transplantation from a 1-of-6 HLA-matched donor. He acquired full enteral tolerance and normal liver function and has never shown evidence of allograft rejection. After mild graft-versus-host disease developed, studies revealed that more than 99% of his CD3(+) lymphocytes and 50% of his CD19(+) lymphocytes were of donor origin, whereas granulocytes and monocytes remained of recipient origin. He synthesizes polyclonal immunoglobulin G (IgG), IgA, and IgM and has developed antibodies to cytomegalovirus (CMV) and parainfluenza 3. His T lymphocytes are predominately CD3(+)CD4(-)CD8(-) with T cell receptor gammadelta heterodimers and CD3(+)CD4(-)CD8(+) with CD8alphaalpha homodimers, populations consistent with an intraepithelial lymphocyte phenotypic profile. We postulate that he has engrafted a donor intestine-derived immune system and is incapable of rejecting his engrafted organs. PMID- 14525786 TI - CD44: a new means to inhibit acute myeloid leukemia cell proliferation via p27Kip1. AB - Acute myeloid leukemia (AML) is sustained by the extensive proliferation of leukemic stem and progenitor cells, which give rise to the population of leukemic blasts with defective differentiation and low proliferative capacity. We have recently shown that ligation of CD44, a cell surface molecule present on AML cells, with specific monoclonal antibodies (mAbs) inhibits their proliferation. However, its mechanism has not been investigated yet. Here, using the NB4 cell line as a model of proliferating human AML cells, and the A3D8 mAb to ligate CD44, we show for the first time that CD44 ligation stabilizes the cyclin dependent kinase inhibitor p27(Kip1) (p27) protein, resulting in increased association with cyclin E/Cdk2 complexes and inhibition of their kinase activity. Moreover, using a p27 antisense vector, we provide direct evidence that p27 is the main mediator of cell growth arrest by CD44. CD44 ligation also leads to p27 accumulation in THP-1, KG1a, and HL60 cell lines and in primary leukemic cells, suggesting that this process is general in AML. Taken together, our present results suggest that CD44 is a new and efficient means to increase the expression of p27 in AML cells. Considering that elevated expression of p27 is a factor of good prognosis in AML, these results provide a new basis for developing CD44 targeted therapy in AML. PMID- 14525787 TI - In vitro and in vivo evidence of PNH cell sensitivity to immune attack after nonmyeloablative allogeneic hematopoietic cell transplantation. AB - It has been proposed that paroxysmal nocturnal hemoglobinuria (PNH) cells may proliferate through their intrinsic resistance to immune attack. To evaluate this hypothesis, we examined the impact of alloimmune pressure on PNH and normal cells in the clinical setting of nonmyeloablative allogeneic hematopoietic cell transplantation (HCT). Five patients with severe PNH underwent HCT from an HLA matched family donor after conditioning with cyclophosphamide and fludarabine. PNH neutrophils (CD15(+)/CD66b(-)/CD16(-)) were detected in all patients at engraftment, but they subsequently declined to undetectable levels in all cases by 4 months after transplantation. To test for differences in susceptibility to immune pressure, minor histocompatibility antigen (mHa)-specific T-cell lines or clones were targeted against glycosylphosphatidylinositol (GPI)-negative and GPI positive monocyte and B-cell fractions purified by flow cytometry sorting. Equivalent amounts of interferon-gamma (IFN-gamma) were secreted following coculture with GPI-negative and GPI-positive targets. Furthermore, mHa-specific T cell lines and CD8(+) T-cell clones showed similar cytotoxicity against both GPI positive and GPI-negative B cells. Presently, all 5 patients survive without evidence of PNH 5 to 39 months after transplantation. These in vitro and in vivo studies show PNH cells can be immunologically eradicated following nonmyeloablative HCT. Relative to normal cells, no evidence for a decreased sensitivity of PNH cells to T-cell-mediated immunity was observed. PMID- 14525788 TI - Erythropoietin regulates endothelial progenitor cells. AB - Circulating bone marrow-derived endothelial progenitor cells (EPCs) promote vascular reparative processes and neoangiogenesis, and their number in peripheral blood correlates with endothelial function and cardiovascular risk. We tested the hypothesis that the cytokine erythropoietin (EPO) stimulates EPCs in humans. We studied 11 patients with renal anemia and 4 healthy subjects who received standard doses of recombinant human EPO (rhEPO). Treatment with rhEPO caused a significant mobilization of CD34(+)/CD45(+) circulating progenitor cells in peripheral blood (measured by flow cytometry), and increased the number of functionally active EPCs (measured by in vitro assay) in patients (week 2, 312% +/- 31%; week 8, 308% +/- 40%; both P <.01 versus baseline) as well as in healthy subjects (week 8, 194% +/- 15%; P <.05 versus baseline). The effect on EPCs was already observed with an rhEPO dose of about 30 IU/kg per week. Administration of rhEPO increased the number of functionally active EPCs by differentiation in vitro in a dose-dependent manner, assessed in cell culture and by tube formation assay. Furthermore, rhEPO activates the Akt protein kinase pathway in EPCs. Erythropoietin increases the number of functionally active EPCs in humans. Administration of rhEPO or EPO analogs may open new therapeutic strategies in regenerative cardiovascular medicine. PMID- 14525789 TI - Induction of caspase-dependent programmed cell death in B-cell chronic lymphocytic leukemia by anti-CD22 immunotoxins. AB - B cells of chronic lymphocytic leukemia (CLL) are long-lived in vivo, possibly because of defects in apoptosis. We investigated BL22, an immunotoxin composed of the Fv portion of an anti-CD22 antibody fused to a 38-kDa Pseudomonas exotoxin-A fragment. B cells from 22 patients with CLL were immunomagnetically enriched (96% purity) and were cultured with BL22 or an immunotoxin that does not recognize hematopoietic cells. The antileukemic activity of BL22 was correlated with CD22 expression, as determined by flow cytometry. BL22 induced caspase-9 and caspase-3 activation, poly(adenosine diphosphate [ADP]-ribose)polymerase (PARP) cleavage, DNA fragmentation, and membrane flipping. Cell death was associated with the loss of mitochondrial membrane potential and the down-regulation of Mcl-1 and X chromosomal inhibitor of apoptosis protein (XIAP). Furthermore, BL22 induced a proapoptotic 18-kDa Bax protein and conformational changes of Bax. Z-VAD.fmk abrogated apoptosis, confirming that cell death was executed by caspases. Conversely, interleukin-4, a survival factor, inhibited spontaneous death in culture but failed to prevent immunotoxin-induced apoptosis. BL22 cytotoxicity was markedly enhanced when combined with anticancer drugs including vincristine. We also investigated HA22, a newly engineered immunotoxin, in which BL22 residues are mutated to improve target binding. HA22 was more active than BL22. In conclusion, these immunotoxins induce caspase-mediated apoptosis involving mitochondrial damage. Combination with chemotherapy is expected to improve the efficacy of immunotoxin treatment. PMID- 14525790 TI - Presence of functional dendritic cells in patients chronically infected with hepatitis C virus. AB - The absence of expanded numbers of hepatitis C virus (HCV)-reactive CD8(+) T lymphocytes (CTLs) in patients chronically infected with HCV has led to the investigation of dendritic cell (DC) function in this population as a potential cause for this defect. Several studies have shown evidence for impaired monocyte derived DCs in chronically infected patients. As it is difficult to reconcile these data with the fact that patients with chronic HCV are immune competent, we re-evaluated this finding, carefully assessing phenotypic markers and functional activity of patient DCs as compared with noninfected controls. In contrast to these prior studies, DCs from 13 of 13 chronic HCV patients expressed typical maturation markers. These mature DCs were capable of priming allogeneic T lymphocytes, as well as stimulating influenza-specific memory T cells. This finding is consistent with clinical and immunologic data that the deficit in the patient's immune repertoire is HCV-specific and suggests that refined models are required for understanding the role of DCs in HCV pathogenesis. PMID- 14525791 TI - Single administration of stem cell factor, FLT-3 ligand, megakaryocyte growth and development factor, and interleukin-3 in combination soon after irradiation prevents nonhuman primates from myelosuppression: long-term follow-up of hematopoiesis. AB - Preservation of hematopoietic stem and progenitor cell survival is required for recovery from radiation-induced myelosuppression. We recently showed that short term injection of antiapoptotic cytokine combinations into mice soon after lethal gamma irradiation promoted survival. The present study investigated the hematopoietic response of cynomolgus monkeys to a single dose of stem cell factor, FLT-3 ligand, megakaryocyte growth and development factor, and interleukin-3 in combination (4F, each factor given intravenously at 50 microg/kg) administered 2 hours after 5-Gy gamma irradiation. Treated monkeys (n = 4) experienced no thrombocytopenia. Only 1 in 4 displayed a transient period of neutropenia (neutrophil [ANC] count < 0.5 x 10(9)/L), whereas all irradiated controls (n = 4) experienced neutropenia (5-12 days) and thrombocytopenia (platelet [PLT] count < 20 x 10(9)/L, 5-31 days). Treated animals exhibited an impressive 2-wave PLT response that peaked at days 8 and 22 after total body irradiation (TBI). Areas under the curve (AUC) of PLTs, ANCs, white blood cells (WBCs), and red blood cells (RBCs) between days 0 and 90 were significantly higher in treated animals than in controls. Humeral bone marrow-derived clonogenic activity was significantly spared at 24 hours and 4 days after TBI in treated monkeys. No apparent impairment of the hematopoietic status and stem cell pool, in terms of long-term culture-initiating cells (LTC-ICs) and side population (SP) cells, was observed after 15 months. These results strongly suggest that the 4F cytokine combination, as a single dose regimen, could act as an emergency treatment for nuclear accident or terrorism victims. PMID- 14525792 TI - The natural history of respiratory syncytial virus infection in cancer and transplant patients: implications for management. AB - Respiratory syncytial virus (RSV) has been reported to cause severe morbidity and mortality among cancer patients receiving chemotherapy with or without autologous peripheral blood stem cell transplantation (APBSCT). However, little is known about the natural history of this infection in these patients, and current standard practice, aerosolized ribavirin plus intravenous immunoglobulin (IVIG), is extremely expensive, difficult to use, and not supported by controlled clinical trials. The purpose of this observational study was to determine the frequency, seasonality, morbidity, and mortality of RSV infection in a group of cancer patients receiving cytotoxic chemotherapy with neither ribavirin nor IVIG treatment. During the period of October 3, 1997, through October 14, 1998, 190 cancer patients (median age, 58 years; 71 women) underwent viral nasopharyngeal washing prior to chemotherapy. Multiple myeloma (MM) accounted for most patients (147, 77%). RSV was recovered from cultures taken from 71 patients (37%) throughout the year, although more frequently during fall and winter seasons (P <.001) than spring and summer. Serious respiratory complications developed in 19 (27%) of 71 RSV-positive patients versus 24 (20%) of 119 patients whose RSV cultures were negative (P =.384). The presence of renal failure or increased lactate dehydrogenase (LDH) prior to chemotherapy and the development of mucositis were the only predictive factors for severe respiratory complications. Recovery of RSV from nasopharyngeal washings among cancer patients is common, occurs throughout the year, and does not appear to increase serious morbidity or mortality. RSV infection may not necessarily be a contraindication for APBSCT or an indication for therapy with aerosolized ribavirin and IVIG. PMID- 14525793 TI - An amino acid polymorphism in von Willebrand factor correlates with increased susceptibility to proteolysis by ADAMTS13. AB - The hypothesis that increased ADAMTS13 (a disintegrin and metalloprotease with thrombospondin repeats) activity or increased susceptibility of von Willebrand factor (VWF) to proteolysis by ADAMTS13 may underlie type I von Willebrand disease (VWD) in some patients was investigated. Plasma from 4 patients with type I VWD was cryoprecipitated. ADAMTS13 activity in the VWF-poor cryodepleted fraction was assessed by incubation with purified VWF; susceptibility to proteolysis of the VWF in the VWF-rich cryoprecipitate was assessed by incubation with a normal, group O cryodepleted plasma. ADAMTS13 activity was similar in all 4 type I VWD cryodepleted plasmas and comparable to a normal control plasma. In contrast, the VWF of one patient showed increased susceptibility to proteolysis by ADAMTS13. Investigation of additional family members indicated that increased susceptibility was heritable, but it did not track uniquely with type I VWD. Sequence analysis of VWF exon 28 indicated that increased susceptibility to proteolysis tracked with the "G" allele of the A/G polymorphism at position 24/1282, encoding the amino acid polymorphism Tyr/Cys1584 ("G" = Cys1584). A prospective study of 200 individuals yielded 2 Tyr/Cys1584 heterozygotes; for both, plasma VWF showed increased susceptibility to proteolysis. The finding that an amino acid polymorphism in VWF may influence susceptibility to ADAMTS13 has potentially significant implications in diverse areas. PMID- 14525794 TI - Regulation of superoxide anion production by NADPH oxidase in monocytes/macrophages: contributions to atherosclerosis. AB - Monocyte extravasation into the vessel wall has been shown to be a critical step in the development of atherosclerosis. Upon activation, monocytes produce a burst of superoxide anion due to activation of the NADPH oxidase enzyme complex. Monocyte-derived superoxide anion contributes to oxidant stress in inflammatory sites, is required for monocyte-mediated LDL oxidation, and alters basic cell functions such as adhesion and proliferation. We hypothesize that monocyte derived superoxide anion production contributes to atherosclerotic lesion formation. In this brief review, we summarize our current understanding of the signal transduction pathways regulating NADPH oxidase activation and related superoxide anion production in activated human monocytes. Novel pathways are identified that may serve as future targets for therapeutic intervention in this pathogenic process. The contributions of superoxide anion and NADPH oxidase to atherogenesis are discussed. Future experiments are needed to clarify the exact role of NADPH oxidase-derived superoxide anion in atherogenesis, particularly that derived from monocytes. PMID- 14525795 TI - Vascular endothelial growth factor-regulated gene expression in endothelial cells: KDR-mediated induction of Egr3 and the related nuclear receptors Nur77, Nurr1, and Nor1. AB - OBJECTIVE: The program of gene expression regulated by vascular endothelial growth factor (VEGF) remains poorly understood. The aim of this study was to identify VEGF-regulated genes in human umbilical vein endothelial cells. METHODS AND RESULTS: VEGF-regulated gene expression was analyzed by screening Affymetrix oligonucleotide arrays and quantitative, real-time, reverse transcription polymerase chain reaction. The most strongly induced genes were the NR4A nuclear receptor family members Nur77, Nurr1, and Nor1 and the zinc-finger transcription factor Egr3. VEGF also induced rapid expression of Down syndrome candidate region 1, cyclooxygenase-2, tissue factor, stanniocalcin-1, the serine/threonine kinase Cot, and eps15 homology domain-containing protein. VEGF-induced NR4A family and Egr3 expression was blocked by a KDR inhibitor, and placental growth factor and basic fibroblast growth factor weakly increased expression of these genes. Induction of NR4A genes was mediated via intracellular Ca2+, protein kinase C- and calcineurin-dependent pathways. VEGF increased protein expression of Nurr1 and Nur77 and decreased Nur77 phosphorylation at the negative regulatory site serine 351. CONCLUSIONS: VEGF induces expression of NR4A nuclear receptors and Egr3 via KDR and KDR-mediated signaling mechanisms. The genes identified here are novel candidates as key early mediators of VEGF-induced endothelial functions. PMID- 14525796 TI - Coronary vessel development: a unique form of vasculogenesis. AB - Development of the coronary vascular system is an interesting model in developmental biology with major implications for the clinical setting. Although coronary vessel development is a form of vasculogenesis followed by angiogenesis, this system uses several unique developmental processes not observed in the formation of other blood vessels. This review summarizes the literature that describes the development of the coronary system, highlighting the unique aspects of coronary vessel development. It should be noted that many of the basic mechanisms that govern vasculogenesis in other systems have not been analyzed in coronary vessel development. In addition, we present recent advances in the field that uncover the basic mechanisms regulating the generation of these blood vessels and identify areas in need of additional studies. PMID- 14525797 TI - Tissue angiotensin-converting-enzyme (ACE) deficiency leads to a reduction in oxidative stress and in atherosclerosis: studies in ACE-knockout mice type 2. AB - Background- Angiotensin II, produced by angiotensin-converting-enzyme (ACE), enhances oxidative stress and atherogenesis. In this study, we analyzed whether tissue ACE deficiency in ACE-knockout mice type-2 would affect their oxidative status. Moreover, by crossbreeding the ACE-knockout mice with atherosclerotic apolipoprotein E (apo E)-deficient (E0) mice, we questioned whether tissue ACE deficiency affects atherogenesis. METHODS AND RESULTS: ACE-deficient mice type-2 (ACE+/-) exhibited reduced serum lipid peroxidation compared with ACE+/+ mice. Peritoneal macrophages from ACE+/- mice demonstrated lower oxidative status, as exhibited by decreases of 47%, 33% 56%, and 51%, in their lipid peroxides, superoxide release, dichlorofluorescein fluorescence, and LDL oxidation, respectively, compared with ACE+/+ mice. ACE+/- mice crossbred with E0 mice, resulting in atherosclerotic mice heterozygous for ACE (ACE+/-/E0 mice), exhibited reduced lipid peroxidation, increased paraoxonase activity, and lower macrophage LDL oxidation compared with E0 and ACE+/+/E0 mice. ACE+/-/E0 mice also exhibited reduced NADPH-induced aortic superoxide ion production by 52% and a reduction of 43% in their atherosclerotic lesion size compared with E0 mice. Finally, 2 animals genotyped as homozygous-knockout for both ACE and APOE genes (ACE-/-/E0), exhibited a striking reduction of 86% in their atherosclerotic lesion area compared with E0 mice. CONCLUSIONS: Reduction of tissue ACE with the ACE-knockout mouse type-2 model inhibited oxidative stress and atherogenesis. PMID- 14525798 TI - Inhaled corticosteroids in chronic obstructive pulmonary disease and risk of death and hospitalization: time-dependent analysis. AB - Observational studies of inhaled corticosteroids in chronic obstructive pulmonary disease (COPD) have shown improved survival whereas randomized trials have not. It has been suggested that this difference may be due to immortal time bias. To investigate this further, we performed a prospective cohort study of patients with COPD, using time-dependent methods to determine whether use of inhaled corticosteroids more than 80% of the time reduced the risk of all-cause mortality and COPD exacerbations. Of 8,033 patients, 2,686 (33%) received inhaled corticosteroids. We did not find a significant reduction in mortality for average inhaled steroid use at either low (hazard ratio [HR], 0.96; 95% confidence interval [CI], 0.69-1.33) or medium/high dose (HR, 0.86; 95% CI, 0.67-1.10). Similarly, recent inhaled corticosteroid use was not associated with a reduction in mortality at low (HR, 0.80; 95% CI, 0.60-1.07) or medium/high doses (HR, 0.88; 95% CI, 0.71-1.09). There was no association between inhaled corticosteroid use and hospitalizations or exacerbations due to COPD. Patients using medium/high dose inhaled corticosteroids did not have a significantly lower risk of COPD hospitalizations (HR, 0.85; 95% CI, 0.64-1.13) or COPD exacerbations (HR, 1.13; 95% CI, 0.94-1.36). In a time-dependent study of outpatients with COPD, adherence to inhaled corticosteroid use was not associated with a decreased risk of mortality or exacerbations. PMID- 14525799 TI - Ventilatory response to brief arousal from non-rapid eye movement sleep is greater in men than in women. AB - Sleep apnea syndromes are more common in men than in women. The ventilatory response to arousal from sleep may be an important determinant of respiratory stability/instability and could contribute to this sex difference. We therefore compared changes in ventilation, end-tidal carbon dioxide (CO2), upper airway resistance, heart rate, and finger photoplethysmogram pulse wave amplitude after both spontaneous and tone-induced arousal from non-rapid eye movement sleep in 13 men and 13 women. At sleep onset, ventilation fell and both upper airway resistance and end-tidal CO2 rose, but these changes were not different between sexes. Spontaneous arousal (duration, 6.6 +/- 0.2 seconds) resulted in a biphasic ventilatory response consisting of brief hyperventilation (5 seconds) followed by prolonged hypoventilation (30-40 seconds) on resumption of sleep. The biphasic ventilatory response was greater in men than in women and did not appear to be explained by different wake-to-sleep increments in end-tidal CO2 or upper airway resistance between sexes. Peripheral vasoconstriction with arousal was also greater in men than in women. Ventilatory responses were more marked after tone induced versus spontaneous arousals and when subjects slept supine compared with the left lateral position. These results suggest that male sex and supine position are associated with greater ventilatory instability after arousal from sleep. PMID- 14525800 TI - Roles of FcgammaRIIB in nasal eosinophilia and IgE production in murine allergic rhinitis. AB - The low-affinity IgG Fc receptor, FcgammaRIIB, displays inhibitory potential in experimental models such as autoimmune diseases. However, whether this receptor is involved in the onset of allergic diseases remains unknown. This study examines the role of FcgammaRIIB in the initiation of allergic rhinitis in mice. Repeated intranasal sensitization with Schistosoma mansoni egg antigen (SEA) induced SEA-specific IgE and marked nasal eosinophilia in high-responder BALB/c mice. FcgammaRIIB gene-deficient (-/-) BALB/c mice displayed severe eosinophilia compared with that of wild-type counterparts. However, FcgammaRIIB -/- mice conversely produced less SEA-specific IgE. The production of interleukin (IL)-4 but not of IL-5 or IFN-gamma by nasal mononuclear cells was also decreased in FcgammaRIIB -/- mice, suggesting that the exacerbation of nasal eosinophila in FcgammaRIIB -/- mice is independent of the local IL-5 levels. The findings in low responder C57BL/6 mice were similar. In addition, nasal eosinophilia in FcgammaRIIB -/- mice passively sensitized with SEA was exacerbated, and conversely, specific IgE production was inhibited after a nasal challenge. These results suggest that FcgammaRIIB plays a regulatory role in the initiation of allergic rhinitis that is independent of either mouse strain or type of sensitization. PMID- 14525801 TI - A combination of oral sildenafil and beraprost ameliorates pulmonary hypertension in rats. AB - Sildenafil, an oral phosphodiesterase type-5 inhibitor, has vasodilatory effects through a cyclic guanosine 3', 5'-monophosphate-dependent mechanism, whereas beraprost, an oral prostacyclin analog, induces vasorelaxation through a cAMP dependent mechanism. We investigated whether the combination of oral sildenafil and beraprost is superior to each drug alone in the treatment of pulmonary hypertension. Rats were randomized to receive repeated administration of saline, sildenafil, beraprost, or both of these drugs twice a day for 3 weeks. Three weeks after monocrotaline (MCT) injection, there was significant development of pulmonary hypertension. The increases in right ventricular systolic pressure and ratio of right ventricular weight to body weight were significantly attenuated in the Sildenafil and Beraprost groups. Combination therapy with sildenafil and beraprost had additive effects on increases in plasma cAMP and cyclic guanosine 3', 5'-monophosphate levels, resulting in further improvement in pulmonary hemodynamics compared with treatment with each drug alone. Unlike MCT rats given saline, sildenafil, or beraprost alone, all rats treated with both drugs remained alive during 6-week follow-up. These results suggest that combination therapy with oral sildenafil and beraprost attenuates the development of MCT-induced pulmonary hypertension compared with treatment with each drug alone. PMID- 14525802 TI - The long-term psychological effects of daily sedative interruption on critically ill patients. AB - Critically ill patients often receive sedatives, which may delay liberation from mechanical ventilation and intensive care unit discharge. Daily interruption of sedatives alleviates these problems, but the impact of this practice on long-term psychological outcomes is unknown. We compared psychological outcomes of intensive care unit patients undergoing daily sedative interruption (intervention) with those without this protocol (control). Assessments using (1) the Revised Impact of Event Scale (evaluates signs of posttraumatic stress disorder [PTSD]), (2) the Medical Outcomes Study 36 item short-form health survey, (3) the State-Trait Anxiety Inventory, (4) the Beck Depression Inventory 2, (5) and the Psychosocial Adjustment to Illness score (overall quality of adjustment to current or residual effects of illness) were done by blinded observers. The intervention group had a better total Impact of Events score (11.2 vs. 27.3, p=0.02), a trend toward a lower incidence of PTSD (0% vs. 32%, p=0.06), and a trend toward a better total Psychosocial Adjustment to Illness score (46.8 vs. 54.3, p=0.08). We conclude that daily sedative interruption does not result in adverse psychological outcomes, reduces symptoms of PTSD, and may be associated with reductions in posttraumatic stress disorder. PMID- 14525803 TI - Reduced interferon gamma production and soluble CD14 levels in early life predict recurrent wheezing by 1 year of age. AB - It is unknown whether reduced production of IFNgamma in early life, before any lower respiratory tract illness, is a risk factor for recurrent wheezing in infancy. We followed 238 infants prospectively from birth to 1 year of age. At birth and at 3 months of age, IFNgamma production from polyclonally stimulated peripheral blood mononuclear cells and soluble CD14 (sCD14) levels in plasma were measured. The odds of developing recurrent wheezing (assessed by questionnaire) in the first year of life were up to 4.5 times higher for children in the lowest quartile of IFNgamma production at 3 months (p = 0.0005) and 3.2 times higher for children in the lowest quartile of sCD14 levels at birth (p = 0.004) as compared with children in the other 3 combined quartiles of IFNgamma and sCD14, respectively. Findings were confirmed in the multivariate analysis. IFNgamma production at 3 months and sCD14 levels at birth were correlated (r = 0.188, p = 0.031). Our findings from a longitudinal cohort suggest that impaired IFNgamma production at 3 months and reduced plasma-sCD14 levels at birth significantly increase the risk of developing recurrent wheezing in the first year of life. PMID- 14525804 TI - Randomized short-term trial of two autoCPAP devices versus fixed continuous positive airway pressure for the treatment of sleep apnea. AB - We evaluated the efficacy of two different continuous positive airway pressure devices with automatic mask pressure adjustment (autoCPAP) in comparison with fixed CPAP in treating obstructive sleep apnea syndrome in 29 patients. The mean (+/- SE) apnea-hypopnea index was 46 +/- 4 per hour and the Epworth score was 14.2 +/- 0.7. Patients were treated over three consecutive 1-month periods with three regimens in random order: an autoCPAP device responding to apnea-hypopnea and snoring, another autoCPAP device responding to snoring and changes in flow contour, and fixed CPAP at the 90th pressure percentile titrated by autoCPAP over 2 weeks. Allowed pressure in the autoCPAP mode was 4 to 15 cm H2O. At the end of each treatment period, symptoms, quality of life, vigilance, and nocturnal breathing disturbances were evaluated. All three treatment modalities improved symptoms, quality-of-life domains, and apnea-hypopnea index significantly and to a similar degree. Mean (+/- SE) maintenance-of-wakefulness time increased by 4.5 +/- 1.8, 6.0 +/- 1.5, and 6.1 +/- 1.4 minutes with DeVilbiss AutoAdjust LT, AutoSet T, and fixed-pressure CPAP, respectively (p<0.001 vs. baseline, p=not significant for comparisons among the three modalities). We conclude that both autoCPAP devices were equally effective as fixed-pressure CPAP in improving major outcomes during short-term therapy of sleep apnea. PMID- 14525806 TI - Increased upper airway collapsibility in children with obstructive sleep apnea during wakefulness. AB - Upper airway collapsibility (UAC) is increased in children with sleep-disordered breathing (SDB), but during wakefulness, active neural processes preserve upper airway patency, such that measurement of upper airway dynamics using acoustic pharyngometry may contribute to diagnostic accuracy in snoring children. Upper airway cross-sectional area obtained from acoustic pharyngometry measurements was assessed in 247 children referred for evaluation of suspected SDB and control subjects, before and after application of cetacaine 1% spray to the pharyngeal introitus under visual inspection. UAC was determined from the precentage change in cross-sectional area after topical anesthesia. UAC measurements were reproducible 1 week apart in both control subjects and patients with SDB (p < 0.005). A UAC less than or equal to -30% exhibited high sensitivity and specificity in identification of all children with obstructive apnea-hypopnea index greater than 5/hour total sleep time in a prospective initial sample of 54 children and in a subsequent post hoc sample of 94 snoring children. Thus, upper airway dynamic testing during wakefulness in response to a topical airway anesthetic may provide a useful clinical adjunct to the evaluation of snoring children, with more accurate identification of those children with SDB. PMID- 14525807 TI - Rho-associated protein kinase contributes to early atherosclerotic lesion formation in mice. AB - Members of the Rho family of small GTPases have been recently implicated in inflammatory signaling. We examined the effect of in vivo inhibition of Rho kinase on atherogenesis in mice. Low-density lipoprotein receptor (LDLR) knockout (KO) mice fed a cholate-free high-fat diet received daily intraperitoneal injection of saline (n=8, control group) or Y-27632 (30 mg/kg, n=9), a specific Rho kinase inhibitor. After 9 weeks, Y-27632 treatment resulted in significant in vivo inhibition of Rho kinase activity (P=0.004). Body weights, arterial blood pressures, and plasma cholesterol levels were comparable in both groups. Atherosclerotic lesion size in the aortic sinus and thoracic aorta of mice treated with Y-27632 was reduced by respectively 35% and 29% in comparison with the saline-treated animals (P=0.006 and P=0.03, respectively). This was associated with a significant reduction in T lymphocyte accumulation (P=0.035) and expression of p65 subunit of NF-kappaB within plaques (P<0.05). In vitro, treatment with Y-27632 inhibited p65 phosphorylation and degradation of IkappaBalpha in mouse peritoneal macrophages and significantly inhibited concanavalin A-induced proliferation of spleen-derived T cells (P<0.001). In conclusion, inhibition of Rho kinase significantly limits early atherosclerotic plaque development in the LDLR KO mice. This study identifies Rho kinase inhibitors as potential candidates for the treatment of atherosclerosis. PMID- 14525809 TI - Neuropeptide Y is an essential in vivo developmental regulator of cardiac ICa,L. AB - Cell culture studies demonstrate an increase in cardiac L-type Ca2+ current (ICa,L) density on sympathetic innervation in vitro and suggest the effect depends on neurally released neuropeptide Y (NPY). To determine if a similar mechanism contributes to the postnatal increase in ICa,L in vivo, we prepared isolated ventricular myocytes from neonatal and adult mice with targeted deletion of the NPY gene (Npy-/-) and matched controls (Npy+/+). Whole-cell voltage clamp demonstrates ICa,L density increases postnatally in Npy+/+ (by 56%), but is unchanged in Npy-/-. Both ICa,L density and action potential duration are significantly greater in adult Npy+/+ than Npy-/- myocytes, whereas ICa,L density is equivalent in neonatal Npy+/+ and Npy-/- myocytes. These data indicate NPY does not influence ICa,L prenatally, but the postnatal increase in ICa,L density is entirely NPY-dependent. In contrast, there is a similar postnatal negative voltage shift in the I-V relation in Npy+/+ and Npy-/-, indicating NPY does not influence the developmental change in ICa,L voltage-dependence. Immunoblot analyses and measurements of maximally activated ICa,L (in presence of forskolin or BayK 8644) show that the differences in current density between Npy+/+ and Npy /- cannot be attributed to altered Ca2+ channel alpha1C subunit protein expression. Rather, these results suggest that the in vivo NPY-dependent postnatal increase in ICa,L density in cardiac myocytes results from regulation ICa,L properties by NPY. PMID- 14525810 TI - Bone marrow monocyte lineage cells adhere on injured endothelium in a monocyte chemoattractant protein-1-dependent manner and accelerate reendothelialization as endothelial progenitor cells. AB - Peripheral blood (PB)-derived CD14+ monocytes were shown to transdifferentiate into endothelial cell (EC) lineage cells and contribute to neovascularization. We investigated whether bone marrow (BM)- or PB-derived CD34-/CD14+ cells are involved in reendothelialization after carotid balloon injury. Although neither hematopoietic nor mesenchymal stem cells were included in human BM-derived CD34 /CD14+ monocyte lineage cells (BM-MLCs), they expressed EC-specific markers (Tie2, CD31, VE-cadherin, and endoglin) to an extent identical to mature ECs. When BM-MLCs were cultured with vascular endothelial growth factors, hematopoietic markers were drastically decreased and new EC-specific markers (Flk and CD34) were induced. BM-MLCs were intra-arterially transplanted into balloon injured arteries of athymic nude rats. When BM-MLCs were activated by monocyte chemoattractant protein-1 (MCP-1) in vivo or in vitro, they adhered onto injured endothelium, differentiated into EC-like cells by losing hematopoietic markers, and inhibited neointimal hyperplasia. Ability to prevent neointimal hyperplasia was more efficient than that of BM-derived CD34+ cells. MCP-dependent adhesion was not observed in PB-derived CD34-/CD14+ monocytes. Regenerated endothelium exhibited a cobblestone appearance, blocked extravasation of dye, and induced NO dependent vasorelaxation. Basal adhesive activities on HUVECs under laminar flow and beta1-integrin expression (basal and active forms) were significantly increased in BM-MLCs compared with PB-derived monocytes. MCP-1 markedly enhanced adhesive activity of BM-MLCs (2.8-fold) on HUVECs by activating beta1-integrin conformation. Thus, BM-MLCs can function as EC progenitors that are more potent than CD34+ cells and acquire the ability to adhere on injured endothelium in a MCP-1-dependent manner, leading to reendothelialization associated with inhibition of intimal hyperplasia. This will open a novel window to MCP-1 mediated biological actions and vascular regeneration strategies by cell therapy. PMID- 14525808 TI - Paracrine regulation of angiogenesis and adipocyte differentiation during in vivo adipogenesis. AB - With an increasing incidence of obesity worldwide, rational strategies are needed to control adipogenesis. Growth of any tissue requires the formation of a functional and mature vasculature. To gain mechanistic insight into the link between active adipogenesis and angiogenesis, we developed a model to visualize noninvasively and in real time both angiogenesis and adipogenesis using intravital microscopy. Implanted murine preadipocytes induced vigorous angiogenesis and formed fat pads in a mouse dorsal skin-fold chamber. The newly formed vessels subsequently remodeled into a mature network consisting of arterioles, capillaries, and venules, whereas the preadipocytes differentiated into adipocytes as confirmed by increased aP2 expression. Inhibition of adipocyte differentiation by transfection of preadipocytes with a peroxisome proliferator activated receptor gamma dominant-negative construct not only abrogated fat tissue formation but also reduced angiogenesis. Surprisingly, inhibition of angiogenesis by vascular endothelial growth factor receptor-2 (VEGFR2) blocking antibody not only reduced angiogenesis and tissue growth but also inhibited preadipocyte differentiation. We found that part of this inhibition stems from the paracrine interaction between endothelial cells and preadipocytes and that VEGF-VEGFR2 signaling in endothelial cells, but not preadipocytes, mediates this process. These findings reveal a reciprocal regulation of adipogenesis and angiogenesis, and suggest that blockade of VEGF signaling can inhibit in vivo adipose tissue formation. The full text of this article is available online at http://www.circresaha.org. PMID- 14525811 TI - Learning from others. PMID- 14525812 TI - Should a normal thromboelastogram allow us to perform a neuraxial block? A strong word of warning. PMID- 14525813 TI - Best evidence in anesthetic practice: prevention: fondaparinux is better than enoxaparin for prevention of major venous thromboembolism after orthopedic surgery. PMID- 14525814 TI - Statistical process control methods allow the analysis and improvement of anesthesia care. AB - PURPOSE: Quality aspects of the anesthetic process are reflected in the rate of intraoperative adverse events. The purpose of this report is to illustrate how the quality of the anesthesia process can be analyzed using statistical process control methods, and exemplify how this analysis can be used for quality improvement. METHODS: We prospectively recorded anesthesia-related data from all anesthetics for five years. The data included intraoperative adverse events, which were graded into four levels, according to severity. We selected four adverse events, representing important quality and safety aspects, for statistical process control analysis. These were: inadequate regional anesthesia, difficult emergence from general anesthesia, intubation difficulties and drug errors. We analyzed the underlying process using 'p-charts' for statistical process control. RESULTS: In 65,170 anesthetics we recorded adverse events in 18.3%; mostly of lesser severity. Control charts were used to define statistically the predictable normal variation in problem rate, and then used as a basis for analysis of the selected problems with the following results: Inadequate plexus anesthesia: stable process, but unacceptably high failure rate; Difficult emergence: unstable process, because of quality improvement efforts; Intubation difficulties: stable process, rate acceptable; Medication errors: methodology not suited because of low rate of errors. CONCLUSION: By applying statistical process control methods to the analysis of adverse events, we have exemplified how this allows us to determine if a process is stable, whether an intervention is required, and if quality improvement efforts have the desired effect. PMID- 14525815 TI - Piroxicam gel, compared to EMLA cream is associated with less pain after venous cannulation in volunteers. AB - PURPOSE: To evaluate and compare the analgesic efficacy and anti-inflammatory effects of topical piroxicam gel vs eutectic mixture of local anesthetic (EMLA) cream applied to the peripheral venous cannulation site in adult volunteers. METHODS: Piroxicam gel and EMLA cream were randomly applied on the dorsum of the right and left hand of ten volunteers who acted as their own control. A venous cannula was inserted (no iv infusion) and removed after one hour. Pain scores and signs of inflammation were noted at the cannulation site up to 48 hr. RESULTS: Pain scores with piroxicam gel were higher on cannulation and on advancement of the cannula (P < 0.05). Thereafter, pain scores were significantly higher with EMLA (P < 0.05). Blanching was present at all the peripheral venous sites treated with EMLA cream. Signs of inflammation (erythema, edema) were not more frequent with EMLA than with piroxicam (P > 0.05). Induration was more frequent with EMLA at six hours. CONCLUSION: In volunteers EMLA cream is associated with less pain on cannulation and cannula advancement compared to piroxicam gel. Topical application of piroxicam gel before peripheral venous cannulation alleviates pain and, possibly, inflammation in the period subsequent to cannulation itself. PMID- 14525816 TI - Neuromuscular blockade at the larynx, the diaphragm and the corrugator supercilii muscle: a review. AB - PURPOSE: To review recent findings concerning neuromuscular blockade and monitoring at the larynx, the diaphragm, and the corrugator supercilii muscle. SOURCE: This narrative review is based on recent publications. PRINCIPAL FINDINGS: Neuromuscular blockade at the larynx and the diaphragm is less intense than at the adductor pollicis muscle; the onset and offset of neuromuscular blockade is more rapid. The corrugator supercilii muscle reflects better the time course of neuromuscular blockade of the larynx than the adductor pollicis muscle, is better suited to monitor the onset of neuromuscular blockade for intubation, and should give a better reflection of the time course and degree of neuromuscular blockade of the larynx or the diaphragm. Recovery of neuromuscular function at the end of any procedure is best reflected at the adductor pollicis muscle where neuromuscular transmission is last restored. Clinical monitoring of the larynx or the diaphragm is still limited by the absence of a simple method. Acceleromyography of the corrugator supercilii muscle is prone to artifacts that do not occur during monitoring of the adductor pollicis muscle. Phonomyography, a new method of monitoring that is currently being tested, is based on the phenomenon that muscle contraction creates low-frequency sound waves, which can be detected using special microphones to quantify neuromuscular blockade. This method seems promising because it can be easily used on all muscles of interest. CONCLUSION: Research during the last 15 years has greatly enhanced our knowledge about how muscles react differently to muscle relaxants and has enabled us to achieve better surgical conditions with safer use of muscle relaxants. Interesting technologies have been developed to reliably monitor neuromuscular blockade at the larynx and the diaphragm, but are currently restricted to research settings. Our increased understanding should help us in ongoing efforts to develop the "ideal" muscle relaxant and the "ideal" method of neuromuscular monitoring. PMID- 14525817 TI - Intravenous ropivacaine bolus is a reliable marker of intravascular injection in premedicated healthy volunteers. AB - PURPOSE: We designed the following volunteer study to determine if an intravascular bolus dose of ropivacaine could be found that would reliably produce mild symptoms of central nervous system (CNS) toxicity in sedated humans. METHODS: After Ethics Committee approval and informed consent 15 volunteers were recruited. Cardiovascular (CVS) monitoring including pulse oximetry, electrocardiogram and non-invasive blood pressure monitoring was applied. In phase 1, volunteers received in sequence placebo, 30 mg, 45 mg and 60 mg of ropivacaine as a 10-mL iv bolus over 20 sec with a two-hour rest period between each injection to allow plasma clearance of drug. Volunteers were asked to report symptoms of local anesthetic toxicity on a verbal response scale. After any dose volunteers reporting greater than three symptoms with a severity of > 3/10 for greater than three minutes were excluded from further study doses. The dose that consistently produced mild CNS toxic effects was chosen for phase 2 of the study. In phase 2, volunteers were given iv midazolam 0.03 mg*kg(-1) prior to bolus ropivacaine or placebo in a randomized double-blind crossover fashion. Volunteers were asked to report toxic symptoms and venous blood samples were obtained for ropivacaine assay. RESULTS: In phase 1, ropivacaine 60 mg was found to produce consistent mild symptoms of CNS toxicity. No volunteer experienced major CNS or CVS adverse effect during the study. After midazolam premedication all volunteers reported symptoms with bolus ropivacaine 60 mg. Mean peak ropivacaine venous concentration was 4.48 mg*L(-1). CONCLUSION: An intravascular bolus of ropivacaine 60 mg reliably produces mild CNS toxic symptoms in premedicated volunteers. PMID- 14525818 TI - The anterior combined approach via a single skin injection site allows lower limb anesthesia in supine patients. AB - PURPOSE: Lower limb anesthesia (LLA) requires the combination of, at least, three in-one and sciatic nerve (SCN) blocks. Anterior approaches are easier to perform with minimal discomfort in supine patients, specially for traumatology. Feasibility of a single needle entry combined approach is reported. CLINICAL FEATURES: The combined landmark was applied in 119 ASA I and II patients (32-68 yr) scheduled for surgery below the knee. Needle (nerve stimulation applied through a single 150-mm long b-bevelled insulated needle) was inserted at the midpoint between the two classical approaches. Thirty and 15 mL of 0.5% ropivacaine were injected close to the femoral and the SCN, respectively. During the following 45 min, the extent of sensory block and knee and ankle motor block were assessed. Landmarks were determined within 1.7 min (0.7-2.2 min). The entire procedure was performed within 4.2 min (2.9-7.1 min) from the determination of the landmark to the SCN infiltration. The three-in-one technique was successful in 89.9% while SCN was successful in 94.9%. Femoral and tibial nerves were always blocked. Blockade of the posterior cutaneous femoral nerve was observed in 78% of patients. The extent and the quality of the sensory block always allowed surgery. Additional iv sedation was needed in 32.6% of patients. Motor block (adapted Bromage's scale > 2) was observed in the femoral (98.3%), the obturator (84.8%), the tibial (97.4%) and the common peroneal (85.7%) nerve distributions. No important adverse effects were recorded. CONCLUSION: The anterior combined approach via a single needle entry represents a technically easy and reliable technique to perform LLA in the supine patient. PMID- 14525820 TI - Postdural puncture headache in a parturient with sickle cell disease: use of an epidural colloid patch. AB - PURPOSE: To report the injection of a colloid in the epidural space as an alternative to an epidural blood patch in a woman with sickle cell disease. CLINICAL FEATURES: A Cesarean delivery was performed under spinal anesthesia in a 32-yr-old woman with severe sickle cell disease and a past medical history of vaso-occlusive crisis. In the postoperative period, the patient complained of postdural puncture headache resistant to symptomatic treatment. Because there were no data concerning the safety of a blood patch in this condition, a colloid (a modified fluid gelatin heated to 37 degrees C) was injected in the epidural space instead of blood. Headaches decreased immediately after the epidural injection of the colloid and disappeared totally within 12 hr. CONCLUSION: Data concerning the safety of epidural blood patches in patients with sickle cell disease are lacking. Injection of colloids in the epidural space could be an alternative. PMID- 14525819 TI - The P-glycoprotein inhibitor quinidine decreases the threshold for bupivacaine induced, but not lidocaine-induced, convulsions in rats. AB - PURPOSE: To examine whether inhibition of P-glycoprotein (P-gp) activity by quinidine affects the central nervous system toxicity of lidocaine and racemic bupivacaine (bupivacaine). METHODS: Forty male Sprague-Dawley rats were randomly divided into four groups (n = 10). Fifteen minutes following administration of 15 mg x kg(-1) of quinidine (QL and QB groups) or saline (L and B groups), lidocaine (L and QL groups, 4 mg x kg(-1) x min(-1)) or bupivacaine (B and QB groups, 1 mg x kg(-1) x min(-1)) was infused until convulsions occurred. Concentrations of lidocaine and its primary metabolite, monoethylglycinexylidide (MEGX) and bupivacaine in plasma and in the brain at the onset of convulsions were measured by high-performance liquid chromatography. RESULTS: There were no differences in the dose of lidocaine required to induce convulsions between the L and QL groups. There were no differences in the concentrations of total (L = 17.2 +/- 3.5, QL = 16.6 +/- 2.6 micro g x mL(-1)) or unbound lidocaine (L = 7.8 +/- 2.5, QL = 7.3 +/ 2.3 micro g x mL(-1)), total (L = 1.2 +/- 0.5, QL = 1.3 +/- 0.7 micro g x mL( 1)) or unbound MEGX (L = 0.9 +/- 0.5, QL = 0.8 +/- 0.4 micro g x mL(-1)) in plasma, total lidocaine or MEGX in the brain at the onset of convulsions between the L and QL groups. The dose of bupivacaine required to induce convulsions was comparable in the B and QB groups. At the onset of convulsions, plasma concentrations of both total (B = 4.9 +/- 1.1, QB = 4.0 +/- 0.6 micro g x mL(-1), P = 0.03) and unbound bupivacaine (B = 1.4 +/- 0.6, QB = 0.9 +/- 0.2 micro g x mL(-1), P = 0.02) were significantly lower in the QB group than in the B group. There were no differences in concentration of total bupivacaine in the brain between the B and QB groups. CONCLUSION: These results suggest that quinidine inhibited P-gp activity, resulting in increased brain/plasma concentration ratio of bupivacaine, but not of lidocaine, and decreased the threshold of plasma concentration for bupivacaine-induced convulsions. PMID- 14525821 TI - Anesthesia for pericardial window in a pregnant patient with cardiac tamponade and mediastinal mass. AB - PURPOSE: To present a case report of anesthesia for pericardial window surgery for acute cardiac tamponade in a patient with an anterior mediastinal mass in late pregnancy. CLINICAL FEATURES: A 34-yr-old gravida 2, para 1 patient presented at 29 weeks gestation with dyspnea, orthopnea, chest pain, and cough. Investigations showed an anterior mediastinal mass due to Hodgkin's disease. A course of vinblastine at 31 weeks gestation resulted in symptomatic improvement but at 34 weeks gestation she developed an acute cardiac tamponade for which pericardial window drainage was required. Additional help and equipment were assembled in case of cardiopulmonary deterioration. Intra-arterial pressure and continuous fetal monitoring were established and iv access was secured in both arms and the left foot. After awake fibreoptic intubation, spontaneous ventilation was maintained. Anesthesia consisted of local anesthetic infiltration of the anterior chest wall, supplemented with fentanyl, midazolam, and ketamine. The patient remained stable, was extubated fully awake, and then monitored in an intensive care area. CONCLUSION: This patient presented with acute cardiac tamponade and an anterior mediastinal mass in late pregnancy, an unusual combination of challenges that requires a careful approach to anesthetic management. PMID- 14525822 TI - Amethocaine or ketorolac eyedrops provide inadequate analgesia in pediatric strabismus surgery. AB - PURPOSE: Corrective strabismus surgery is associated with moderate pain after surgery. Postoperative analgesia for these patients may include topical local anesthetic agents and topical non-steroidal anti-inflammatory drugs. In this prospective randomized, double-blind placebo controlled clinical trial we compared the effect of placebo to intraoperative 0.5% topical amethocaine or 0.5% topical ketorolac on pain control after strabismus surgery in children. METHODS: Following Institutional Ethics Committee approval and parental consent, we prospectively studied 51 healthy children between the ages of two and seven years who were undergoing elective bilateral recession surgery in a randomized, double blind controlled clinical trial. Children were randomized to receive either placebo (normal saline), 0.5% amethocaine or 0.5% ketorolac eye drops at the start and end of strabismus repair surgery. Pain was assessed with a modified Children's Hospital of Eastern Ontario Pain Score in the recovery room. If the pain score was greater than 6, the patient was administered a single oral dose of acetaminophen (20 mg x kg(-1)). RESULTS: The groups had similar demographic data. Duration of surgery and anesthesia, time spent in recovery room and length of hospital stay between the three groups were similar. Pain scores and analgesic requirements while in the hospital were also similar between the groups as was the time to first analgesic administration. There were no side effects observed in any of the three treatment arms. CONCLUSION: We conclude that there is no improvement in postoperative pain control after the intraoperative administration of topical 0.5% ketorolac or 0.5% amethocaine when compared to placebo in children undergoing strabismus surgery. PMID- 14525823 TI - Endotracheal intubation with a lightwand or a laryngoscope results in similar hemodynamic variations in patients with coronary artery disease. AB - PURPOSE: To asses the cardiovascular changes after either lightwand or conventional laryngoscopic endotracheal intubation (EI) in patients with coronary artery disease. METHODS: Following Institutional approval and informed consent, 80 consecutive patients undergoing elective coronary artery bypass grafting were enrolled in this prospective, controlled, single-blinded study. General anesthesia was induced with fentanyl 5 micro x kg(-1) and thiopental 5 mg x kg( 1) followed by pancuronium 0.1 mg x kg(-1). After loss-of-eyelash reflex the lungs were manually ventilated with 2% isoflurane in oxygen for five minutes. Patients were then randomly allocated to receive either the lightwand (lightwand group, n = 41) or direct-vision laryngoscopy (laryngoscopy group, n = 39). Heart rate (HR) and direct blood pressure were recorded before induction, after induction but before EI, during EI, immediately after EI and at ten-second intervals for the following five minutes. Hemodynamic management during induction was standardized. Hypotension was treated with volume replacement, ephedrine, or phenylephrine as indicated; hypertension was treated with iv nitroglycerin; tachycardia was treated with boluses of esmolol; and, bradycardia was treated with atropine or ephedrine. RESULTS: In both groups, mean arterial blood pressures and HR increased significantly after EI. There was a tendency for the lightwand group to have lower arterial blood pressures and slower HR. However, the differences between the two groups did not reach statistical significance. Requirements for drugs to control HR and mean arterial pressure were similar in both groups. CONCLUSION: In patients with coronary artery disease using a lightwand intubation technique does not modify the hemodynamic response associated with EI as compared with standard direct-vision laryngoscopy. PMID- 14525824 TI - Images in anesthesia: transesophageal echocardiogram (TEE) images of an anomalous left circumflex coronary artery. PMID- 14525825 TI - Milrinone attenuates the negative inotropic effects of landiolol in halothane anesthetized dogs. AB - BACKGROUND: Clinical use of high dose beta-blocker therapy is limited by excessive negative inotropic effects. Previous studies suggest that milrinone may be of utility in limiting the inotropic but not the chronotropic effects of beta blockers. We examined the hemodynamic effects of co-administration of a new potent selective beta(1) blocker, landiolol, and milrinone in halothane anesthetized dogs. METHODS: Eighteen adult mongrel dogs were anesthetized with 1.2 MAC halothane. Hemodynamic measurements were made at baseline, 30 min after starting the milrinone (0.5 micro g x kg(-1) x min(-1)) or normal saline infusion (n = 9 in each), then 30 min after each change in the dose of landiolol infusion. The tested doses of landiolol were 10, 100, and 1000 micro g x kg(-1) x min(-1). RESULTS: Landiolol (>/= 10 micro g x kg(-1) x min(-1)) has significant and comparable negative chronotropic effects in both groups of dogs. While it also has significant negative inotropic effects in both groups, such effects are significantly attenuated in the dogs treated with milrinone. CONCLUSION: Milrinone is effective to attenuate the negative inotropic effects of landiolol in halothane-anesthetized dogs. PMID- 14525827 TI - Inhaled nitric oxide in 2003: a review of its mechanisms of action. AB - PURPOSE: To review the pulmonary and systemic effects of endogenous nitric oxide and inhaled nitric oxide administered to patients. SOURCE: A systematic search for experimental data, human case reports, and randomized clinical trials since 1980, the year of discovery of endothelium-derived relaxing factor. PRINCIPAL FINDINGS: Nitric oxide has pulmonary and systemic effects. Inhaled nitric oxide not only causes selective pulmonary vasodilation but also results in pulmonary vasoconstriction of the vessels perfusing non-ventilated alveolae. The systemic effects of inhaled nitric oxide, which include modulation of the distribution of systemic blood flow, increase in renal output, interaction with coagulation, fibrinolysis and platelet functions, alteration of the inflammatory response, are described and the mechanisms of nitric oxide transport are explained. The possible toxicity of inhaled nitric oxide is also discussed. CONCLUSION: The multiple effects of inhaled nitric oxide support its role as a pulmonary and extra-pulmonary medication. PMID- 14525826 TI - Venous air embolism during awake craniotomy in a supine patient. AB - PURPOSE: To report a non-fatal case of intraoperative venous air embolism (VAE) during an awake craniotomy. VAE presented with unusual clinical features. CLINICAL FEATURES: VAE during an awake craniotomy has not been reported frequently. The patient we describe presented with persistent coughing followed by tachypnea, hypoxia and reduction in end-tidal CO(2) during dural opening while undergoing an awake craniotomy in the supine position. Cardiovascular variables were stable during the episode except for transient hypertension. Having ruled out airway obstruction and low cardiac output, we concluded that air embolism was the cause. The patient responded immediately to the standard treatment of air embolism and recovered without any complication. CONCLUSION: This case illustrates a VAE during an awake craniotomy and emphasizes the importance of early diagnosis in the management. PMID- 14525828 TI - Futility-of-care decisions in the treatment of moribund intensive care patients in a developing country. AB - PURPOSE: To analyze the characteristics of moribund patients in a surgical intensive care unit (ICU) and highlight the dilemmas inherent in treating such patients. METHODS: Data on all patients admitted to the surgical ICU during the period of three years from July 1999 to June 2002 were collected prospectively. Data were collected on very ill patients who died, in whom it appeared obvious that treatment could not have improved their condition and whose death could have been anticipated. The case notes were subjected to further analysis to determine the difficulties encountered in managing patients whose therapy was considered to be futile. RESULTS: Of 662 admissions, 100 (15.1%) died and 30 (4.5%) patients were treated aggressively, even after a prognosis which reflected futile treatment. The overall mean length of stay for survivors was 7.5 +/- 9.0 [standard deviation (SD)] days and that for the non-survivors was 12.8 +/- 18.1 (SD; P < 0.001). The cost incurred for the treatment of non-survivors was significantly higher than that for the surviving patients. The factors relating to the decisions to continue futile therapy were age of the patient, legal considerations, family wishes and differing opinions between treating physicians. CONCLUSION: Consideration of futility during end-of-life care did not receive adequate attention in this unit which incurred additional human and material resources. PMID- 14525829 TI - Thromboelastography to assess coagulation in the thrombocytopenic parturient. PMID- 14525830 TI - Programming errors from patient-controlled analgesia. PMID- 14525831 TI - AIR tent for airway management of SARS patients. PMID- 14525832 TI - Programming errors from patient-controlled analgesia. PMID- 14525833 TI - Programming errors from patient-controlled analgesia. PMID- 14525835 TI - Programming errors from patient-controlled analgesia. PMID- 14525836 TI - Another use for the forced air warmer. PMID- 14525837 TI - Prophylactic i.v. metaraminol during spinal anesthesia for elective Cesarean delivery. PMID- 14525838 TI - Laryngeal mask airway for preservation of the external branch of the superior laryngeal nerve during thyroid surgery. PMID- 14525839 TI - Acute pulmonary edema in relation with single coronary ostium following acoustic neuroma surgery. PMID- 14525842 TI - Identification of the epidural space: air or saline? PMID- 14525840 TI - Identification of the epidural space: air or saline? PMID- 14525843 TI - Posterior tibial nerve block in the therapeutic management of painful calcaneal spur (plantar fasciitis): a preliminary experience. PMID- 14525844 TI - The effect of right and left lateral position on PA-aO2 and PA-ETCO2 in thoracic surgery. PMID- 14525845 TI - Assessment of neuromuscular blockade at the vastus medialis. PMID- 14525848 TI - Consent to using human tissue. PMID- 14525849 TI - Blindness in children. PMID- 14525850 TI - Treatment of postoperative nausea and vomiting. PMID- 14525851 TI - Race, ethnic group, and clinical research. PMID- 14525852 TI - Small improvement seen in teenager with vCJD. PMID- 14525853 TI - NIH announces new strategy for research. PMID- 14525854 TI - NICE announces guidelines for use of rituximab. PMID- 14525858 TI - Head of German medicines body likens HRT to thalidomide. PMID- 14525859 TI - Doctors test drug to prevent cancer in high risk women. PMID- 14525864 TI - WHO and MSF appeal for funds for new meningitis vaccine. PMID- 14525861 TI - Doctors in India prescribe unapproved fertility drug. PMID- 14525865 TI - No eradication of polio without inactivated vaccine, says expert. PMID- 14525866 TI - Spending on UK health care is increasing faster than other public spending. PMID- 14525869 TI - Medicines committee felt unable to recommend use of pentosan polyphosphate in vCJD. PMID- 14525871 TI - Time trends and demography of mortality after fractured neck of femur in an English population, 1968-98: database study. AB - OBJECTIVES: To investigate time trends in mortality after admission to hospital for fractured neck of femur from 1968 to 1998, and to report on the effects of demographic factors on mortality. DESIGN: Analysis of hospital inpatient statistics for fractured neck of femur, incorporating linkage to death certificates. SETTING: Four counties in southern England. SUBJECTS: 32 590 people aged 65 years or over admitted to hospital with fractured neck of femur between 1968 and 1998. MAIN OUTCOME MEASURES: Case fatality rates at 30, 90, and 365 days after admission, and standardised mortality ratios at monthly intervals up to one year after admission. RESULTS: Case fatality rates declined between the 1960s and the early 1980s, but there was no appreciable fall thereafter. They increased sharply with increasing age: for example, fatality rates at 30 days in 1984-98 increased from 4% in men aged 64-69 years to 31% in those aged > or = 90. They were higher in men than women, and in social classes IV and V than in classes I and II. In the first month after fracture, standardised mortality ratios in women were 16 times higher, and those in men 12 times higher, than mortality in the same age group in the general population. CONCLUSIONS: The high mortality rates, and the fact that they have not fallen over the past 20 years, reinforce the need for measures to prevent osteoporosis and falls and their consequences in elderly people. Whether post-fracture mortality has fallen to an irreducible minimum, or whether further decline is possible, is unclear. PMID- 14525872 TI - Hysterectomy and sexual wellbeing: prospective observational study of vaginal hysterectomy, subtotal abdominal hysterectomy, and total abdominal hysterectomy. AB - OBJECTIVES: To compare the effects of vaginal hysterectomy, subtotal abdominal hysterectomy, and total abdominal hysterectomy on sexual wellbeing. DESIGN: Prospective observational study over six months. SETTING: 13 teaching and non teaching hospitals in the Netherlands. PARTICIPANTS: 413 women who underwent hysterectomy for benign disease other than symptomatic prolapse of the uterus and endometriosis. MAIN OUTCOME MEASURES: Reported sexual pleasure, sexual activity, and bothersome sexual problems. RESULTS: Sexual pleasure significantly improved in all patients, independent of the type of hysterectomy. The prevalence of one or more bothersome sexual problems six months after vaginal hysterectomy, subtotal abdominal hysterectomy, and total abdominal hysterectomy was 43% (38/89), 41% (31/76), and 39% (57/145), respectively (chi2 test, P = 0.88). CONCLUSION: Sexual pleasure improves after vaginal hysterectomy, subtotal abdominal hysterectomy, and total abdominal hysterectomy. The persistence and development of bothersome problems during sexual activity were similar for all three techniques. PMID- 14525873 TI - Dietary fat intake and risk of stroke in male US healthcare professionals: 14 year prospective cohort study. AB - OBJECTIVE: To examine the association between intake of total fat, specific types of fat, and cholesterol and risk of stroke in men. Design and setting Health professional follow up study with 14 year follow up. PARTICIPANTS: 43 732 men aged 40-75 years who were free from cardiovascular diseases and diabetes in 1986. MAIN OUTCOME MEASURE: Relative risk of ischaemic and haemorrhagic stroke according to intake of total fat, cholesterol, and specific types of fat. RESULTS: During the 14 year follow up 725 cases of stroke occurred, including 455 ischaemic strokes, 125 haemorrhagic stokes, and 145 strokes of unknown type. After adjustment for age, smoking, and other potential confounders, no evidence was found that the amount or type of dietary fat affects the risk of developing ischaemic or haemorrhagic stroke. Comparing the highest fifth of intake with the lowest fifth, the multivariate relative risk of ischaemic stroke was 0.91 (95% confidence interval 0.65 to 1.28; P for trend = 0.77) for total fat, 1.20 (0.84 to 1.70; P = 0.47) for animal fat, 1.07 (0.77 to 1.47; P = 0.66) for vegetable fat, 1.16 (0.81 to 1.65; P = 0.59) for saturated fat, 0.91 (0.65 to 1.28; P = 0.83) for monounsaturated fat, 0.88 (0.64 to 1.21; P = 0.25) for polyunsaturated fat, 0.87 (0.62 to 1.22; P = 0.42) for trans unsaturated fat, and 1.02 (0.75 to 1.39; P = 0.99) for dietary cholesterol. Intakes of red meats, high fat dairy products, nuts, and eggs were also not appreciably related to risk of stroke. CONCLUSIONS: These findings do not support associations between intake of total fat, cholesterol, or specific types of fat and risk of stroke in men. PMID- 14525874 TI - Family attitudes to research using samples taken at coroner's postmortem examinations: review of records. PMID- 14525875 TI - British hospitals and different versions of the Glasgow coma scale: telephone survey. PMID- 14525876 TI - Substance misuse in psychiatric inpatients: comparison of a screening questionnaire survey with case notes. PMID- 14525877 TI - Evidence for risk of bias in cluster randomised trials: review of recent trials published in three general medical journals. AB - OBJECTIVE: To examine the prevalence of a risk of bias associated with the design and conduct of cluster randomised controlled trials among a sample of recently published studies. DESIGN: Retrospective review of cluster randomised trials published in the BMJ, Lancet, and New England Journal of Medicine from January 1997 to October 2002. MAIN OUTCOME MEASURES: Prevalence of secure randomisation of clusters, identification of participants before randomisation (to avoid foreknowledge of allocation), differential recruitment between treatment arms, differential application of inclusion and exclusion criteria, and differential attrition. RESULTS: Of the 36 trials identified, 24 were published in the BMJ,11 in the Lancet, and a single trial in the New England Journal of Medicine. At the cluster level, 15 (42%) trials provided evidence for secure allocation and 25 (69%) used stratified allocation. Few trials showed evidence of imbalance at the cluster level. However, some evidence of susceptibility to risk of bias at the individual level existed in 14 (39%) studies. CONCLUSIONS: Some recently published cluster randomised trials may not have taken adequate precautions to guard against threats to the internal validity of their design. PMID- 14525878 TI - Taking simvastatin in the morning compared with in the evening: randomised controlled trial. PMID- 14525879 TI - Diagnostic impact of signs and symptoms in acute infectious conjunctivitis: systematic literature search. PMID- 14525881 TI - Basal cell carcinoma. PMID- 14525880 TI - The Ptolemy project: a scalable model for delivering health information in Africa. PMID- 14525882 TI - Assisted conception. I--General principles. PMID- 14525883 TI - Death of the teaching autopsy. PMID- 14525884 TI - Resuscitating the teaching autopsy. PMID- 14525885 TI - Obtaining consent for autopsy. PMID- 14525886 TI - "Polypill" to fight cardiovascular disease: now who's playing God? PMID- 14525887 TI - "Polypill" to fight cardiovascular disease: universal polypharmacy goes against recent beliefs in prescribing practice. PMID- 14525888 TI - "Polypill" to fight cardiovascular disease: patients before populations. PMID- 14525889 TI - "Polypill" to fight cardiovascular disease: cost effectiveness of statins for primary prevention of cardiovascular events is questionable. PMID- 14525890 TI - "Polypill" to fight cardiovascular disease: old joke has element of truth. PMID- 14525891 TI - "Polypill" to fight cardiovascular disease: interpretation of trial data is optimistic. PMID- 14525892 TI - "Polypill" to fight cardiovascular disease: birthday present was much appreciated. PMID- 14525894 TI - Do school exams predict doctors' success? Is it not a retrospective study? PMID- 14525895 TI - Do school exams predict doctors' success? Career achievements are not only measure. PMID- 14525896 TI - Research was evidence based. PMID- 14525897 TI - Smoke free hospitals: rising to the challenge. PMID- 14525898 TI - Smoke free hospitals: withdrawal from cigarettes should not be confused with withdrawal from nicotine. PMID- 14525899 TI - Dyspepsia results may not apply in primary care. PMID- 14525900 TI - Be careful when extrapolating trial data to real life. PMID- 14525901 TI - Confidentiality of patients' information must be guaranteed. PMID- 14525902 TI - Diabetes may be independent risk factor for hyperkalaemia. PMID- 14525903 TI - A career in respiratory medicine. PMID- 14525904 TI - Specialist registrars acting up as consultants. PMID- 14525905 TI - Courses, cadavers, and counsellors: reducing errors in the operating theatre. PMID- 14525907 TI - Gene therapy for central diabetes insipidus: effective antidiuresis by muscle targeted gene transfer. AB - Central diabetes insipidus, characterized by severe polyuria and polydipsia, is a disorder resulting from deficient secretion of the small neuropeptide hormone vasopressin in the neurohypophysis. The standard therapy is daily and life-long administration of vasopressin analog (desmopressin acetate), but gene therapy is potentially alternative to the conventional replacement therapy. To obtain the therapeutic neuropeptide more feasibly, we tried to express vasopressin in nonneuronal tissues using nonviral gene transfer techniques. We found that the unprocessed large precursor form, provasopressin, was predominantly produced in nonendocrine cells transfected with the wild-type vasopressin gene, because of the lack of neuroendocrine cell-specific endopeptidases. In sharp contrast, appropriately processed bioactive vasopressin can be efficiently produced even in nonendocrine cells with a modified vasopressin gene containing a ubiquitous endoprotease furin cleavage site. We also succeeded in maintaining a long-term antidiuretic effect on vasopressin-deficient (Brattleboro) rats by direct introduction of the furin-processible gene into skeletal muscle by electroporation. Altogether, our data clearly show that skeletal muscle is a useful target tissue for continuous delivery of bioactive neuropeptide. Furthermore, our strategies may be applicable to future gene therapies for central diabetes insipidus and other peptide hormone deficiencies. PMID- 14525906 TI - Osteoprotegerin expression and secretion are regulated by calcium influx through the L-type voltage-sensitive calcium channel. AB - Our previous studies showed that 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] modulates the activity of the Ca(V1.2) alpha-subunit of the L-type voltage-sensitive calcium channel (VSCC) by two temporally distinct mechanisms. First, 1,25(OH)2D3 rapidly modulates local Ca2+ permeability in the plasma membrane of the proliferating osteoblast. Second, treatment with 1,25(OH)2D3 reduces biosynthesis of Ca(V1.2) such that transcript levels are half of original levels after 24 h. Osteoprotegerin (OPG) and receptor activator of nuclear factor kappa B ligand (RANKL) provide important regulatory mechanisms for controlling osteoclastogenesis and Ca2+ homeostasis. Because they often control Ca2+ activated secretion, we investigated the possibility that L-type VSCCs might regulate basal OPG and RANKL secretion in osteoblasts. We also studied 1,25(OH)2D3 effects on OPG and RANKL expression. To address this, we measured changes in expression and secretion of OPG and RANKL in MC3T3-E1 cells and calvarial organ cultures after treatment with 1,25(OH)2D3, VSCC inhibitors, and inhibitors of Ca2+-regulated signaling. RANKL production was increased in calvarial cultures by 1,25(OH)2D3 but was essentially undetectable in the medium of MC3T3-E1 cells. In contrast, OPG secretion in both systems was significantly reduced after 24 h treatment with 1,25(OH)2D3, by inhibitors of L-type VSCCs and calmodulin-sensitive protein kinases but not by inhibitors of protein kinase A, MAPKs, or other families of VSCCs. OPG secretion was abrogated by transfection with decoy cAMP response element binding sites. Our results suggest that OPG secretion is regulated through calmodulin-sensitive protein kinase signaling that depends on the activity of the L-type VSCC and is mediated through the cAMP response element-binding protein. PMID- 14525908 TI - Intravenous 2-deoxy-D-glucose injection rapidly elevates levels of the phosphorylated forms of p44/42 mitogen-activated protein kinases (extracellularly regulated kinases 1/2) in rat hypothalamic parvicellular paraventricular neurons. AB - CRH neurons within the medial parvicellular part of the hypothalamic paraventricular nucleus (PVHmp) can respond to afferent inputs encoding stress related information by initiating peptide synthesis (signaling cascades, transcription, and translation) and/or peptide release. However, understanding these cellular events is hampered by three outstanding issues: 1) neural inputs that activate CRH neurons remain incompletely identified; 2) the identity and temporal dynamics of signaling pathways within CRH neurons are poorly understood; and 3) the precise coupling of the first two issues has not been established. Here, we report that the phosphorylated forms of p44/p42 MAPKs (pERK1/2) are rapidly detected in PVHmp cells after i.v. infusion of the antimetabolite, 2 deoxy-D-glucose (2-DG). Combined immunocytochemistry and in situ hybridization revealed that pERK1/2 immunoreactivity is detectable 10 min after 2-DG infusion not only within most PVHmp neurons containing CRH mRNA (78.6% of mean total CRH cells counted) but also in many non-CRH neurons (45.5% of mean total sampled cells). In contrast, Fos protein in the PVHmp was not detected within this time period, consistent with the known time course for its translation. Stress associated with halothane exposure also robustly elevated pERK1/2 levels in PVHmp neurons approximately 10 min after exposure. Our results implicate pERK1/2 in stress-induced activation of CRH neurosecretory cells and underscore their utility as indices of rapid cellular activation. Because 2-DG-induced activation of CRH gene transcription in these neurons requires a catecholaminergic input, our data also suggest that pERK1/2 could couple afferent catecholaminergic signals with CRH gene expression in these neurons. PMID- 14525909 TI - Regulation of Cbl-associated protein/Cbl pathway in muscle and adipose tissues of two animal models of insulin resistance. AB - The phosphatidylinositol 3-kinase-independent pathway to induce glucose transport may involve the tyrosine phosphorylation of the protooncogene c-Cbl. In the present study, we examined whether acute exposure to insulin stimulates the tyrosine phosphorylation of Cbl and its association with Cbl-associated protein (CAP) in muscle and adipose tissue of rats in vivo. We report herein that insulin induces Cbl tyrosine phosphorylation and association with CAP in adipose tissue but not in muscle. We also examined the expression and tyrosyl-phosphorylation state of Cbl and CAP/Cbl association in adipose tissue of rats submitted to prolonged fasting and in monosodium glutamate (MSG)-insulin-resistant rats. An increase in Cbl phosphorylation is observed in the fat of MSG rats, parallel with an increase in association of CAP-Cbl as well as an augment in CAP and Cbl protein expression in the adipose tissue of these animals. These events are accompanied by a decrease in insulin-stimulated insulin receptor/ insulin receptor substrate (IRS)-1 tyrosine phosphorylation and an increase in the IRS 2/phosphatidylinositol 3-kinase/Akt/Foxo1 pathway. In adipocytes of fasted rats, there is a decrease in CAP and Cbl protein expression, insulin-induced Cbl phosphorylation, and the association with CAP. In parallel, there is also a decrease in the insulin receptor/IRSs/Akt/Foxo1 pathway. Thus, insulin is able to induce Cbl tyrosine phosphorylation and its association with CAP in the adipose tissue of normal rats. In addition, our data provide evidence that the CAP-Cbl pathway may have a role in the modulation of adiposity in fasting and in MSG treated rats. PMID- 14525910 TI - Antiequine chorionic gonadotropin (eCG) antibodies generated in goats treated with eCG for the induction of ovulation modulate the luteinizing hormone and follicle-stimulating hormone bioactivities of eCG differently. AB - In dairy goats, treatments associating a progestogen and the equine chorionic gonadotropin (eCG) are the easiest way to induce and synchronize estrus and ovulation and to permit artificial insemination (AI) and/or out of season breeding. From the first treatment, the injection of eCG induces, in some females, the production of anti-eCG antibodies (Abs) that will interfere with the effectiveness of subsequent treatments. These anti-eCG Abs delay the preovulatory LH surge and the ovulation time, leading to poor fertility of the treated females. In this study, by in vitro bioassays, we show that anti-eCG Abs can positively or negatively modulate the LH and/or FSH bioactivities of eCG. Moreover, the modulation level of eCG bioactivity does not depend on the anti-eCG Ab affinity for eCG, as shown by surface plasmon resonance technology. The specificity of anti-eCG Abs tested by competitive ELISA highlighted the importance of a glycan environment in the recognition mechanism, especially the sialic acids specific to eCG. The different effects of anti-eCG Abs on eCG bioactivities could be explained by two hypotheses. First, steric hindrance preventing the interaction of eCG with its receptors would explain the inhibitory effect of some anti-eCG Abs; second, a conformational change in eCG by anti-eCG Abs could induce inhibition or potentiation of eCG bioactivities. It is significant that these modulations of eCG bioactivities by anti-eCG Abs impact mainly on the FSH bioactivity of eCG, which is essential for ovarian stimulation and subsequent fertility after treatment and AI, and to a lesser extent on LH bioactivity. PMID- 14525911 TI - Effects of 1alpha,25-dihydroxyvitamin D3 and growth hormone on apoptosis and proliferation in UMR 106 osteoblast-like cells. AB - Mechanisms maintaining a correct balance between bone-forming osteoblasts and bone-resorbing osteoclasts are essential for bone formation. Apoptosis has been proposed to play a key role in controlling osteoblast homeostasis. 1,25 dihydroxyvitamin D3 [1,25(OH)2D3] and GH, which are important regulators of bone growth and bone metabolism, also play pivotal roles in regulation of mitogenesis, differentiation, and apoptosis. We have recently shown that 1,25(OH)2D3 prolongs GH signaling via the Janus kinase 2 (JAK2)/STAT5 (signal transducer and activator of transcription 5) pathway in UMR 106 osteoblast-like cells. In the present study, we have investigated the effects of GH and 1,25(OH)2D3 on proliferation and apoptosis in UMR 106 cells. We found that 1,25(OH)2D3 and GH, separate or in combination, inhibited apoptosis. GH also had profound effects on cell cycle distribution and proliferation. In addition, pretreatment of cells with 1,25(OH)2D3 was necessary to detect GH-induced MAPK activation. We hypothesize that these hormones separately regulate the processes of apoptosis and proliferation, which may be important for maintaining osteoblast cell number. PMID- 14525912 TI - Leptin reverses the inhibitory effect of caloric restriction on longitudinal growth. AB - Caloric imbalance, particularly in critical periods of growth and development, is often the underlying cause of growth abnormalities. Serum levels of leptin are elevated in obesity and are low in malnutrition and malabsorption. The aim of the present study was to determine whether leptin integrates energy levels and growth in vivo, as shown previously in our ex vivo experiments, even in the presence of caloric restriction. In the first part of the study, mice were divided into three groups. Two groups were fed ad libitum and received leptin or vehicle only, and the third group was pair-fed with the group injected with leptin to dissociate leptin's effect on growth from its effect on food consumption. Mice given leptin had a significantly greater tibial length than untreated pair-fed animals and a similar tibial length as control mice fed ad libitum despite their lower weight. In addition, leptin significantly increased the overall size of the epiphyseal growth plate by 11%. On immunohistochemistry and in situ hybridization studies, leptin stimulated both the proliferation and differentiation of tibial growth plate chondrocytes without affecting the overall organization of the plate. There was also a marked increase in the expression and level of IGF-IR. In the second part of the study, two groups of mice were fed only 60% of their normal chow; one was injected with leptin, and the other was injected with vehicle alone. Caloric deprivation by itself reduced serum levels of IGF-I by 70% and the length of the tibia by 5%. Leptin treatment corrected the fasting-induced growth deficiency, but further reduced the level of serum IGF-I. These results indicate that leptin stimulates growth even in the presence of caloric restriction independently of peripheral IGF-I. PMID- 14525913 TI - Chronic neuropeptide Y infusion into the lateral ventricle induces sustained feeding and obesity in mice lacking either Npy1r or Npy5r expression. AB - Neuropeptide Y (NPY) is a powerful orexigenic neurotransmitter. The NPY Y1 and Y5 receptors have been implicated in mediating the appetite-stimulating activity of NPY. To further investigate the importance of these two receptors in NPY-induced hyperphagia after chronic central administration, we used mice lacking either Npy1r or Npy5r expression. NPY infusion into the lateral ventricle of wild-type mice stimulated food intake and induced obesity over a 7-d period. Fat pad weight as well as plasma insulin, leptin, and corticosterone levels were strongly increased in NPY-treated mice. In addition, NPY infusion resulted in a significant decrease in hypothalamic NPY and proopiomelanocortin expression. Interestingly, the lack of either Npy1r or Npy5r expression in knockout mice did not affect such feeding response to chronic NPY infusion. Moreover, the obesity syndrome that developed in these animals was similar to that in wild-type animals. Taken together, these data strongly suggest biological redundancies between Y1 and Y5 receptor signaling in the NPY-mediated control of food intake. PMID- 14525914 TI - Vagotomy dissociates short- and long-term controls of circulating ghrelin. AB - Plasma ghrelin levels are responsive to short- and long-term nutrient fluctuation, rapidly decreasing with food consumption and increasing with food deprivation or weight loss. We hypothesized a vagal contribution to both responses. Nutrient-related ghrelin suppression may be mediated by gastrointestinal load-related vagal afferent activity, or depend upon vagal efferent input to the foregut, where most ghrelin is produced. Similarly, the deprivation-induced ghrelin rise could require state-related vagal afferent or efferent activity. Here, we examined the role of the vagus nerve in the regulation of plasma ghrelin by sampling blood from rats with subdiaphragmatic vagotomy and from sham-operated controls over 48 h of food deprivation, and before and after gastric gavage of liquid diet. Vagotomy affected neither baseline ghrelin levels nor the suppression of ghrelin by a nutrient load. The food deprivation-induced elevation of plasma ghrelin levels ( approximately 160% of baseline), however, was completely prevented by subdiaphragmatic vagotomy. In a separate experiment, the deprivation-related rise in plasma ghrelin was substantially reduced by atropine methyl nitrate treatment, indicating that the response to fasting is driven by increased vagal efferent tone. The dissociation between nutrient load- and deprivation-related ghrelin responses indicates that the regulation of circulating ghrelin levels involves separate mechanisms operating through anatomically distinct pathways. PMID- 14525915 TI - The volume of a sexually dimorphic nucleus in the ovine medial preoptic area/anterior hypothalamus varies with sexual partner preference. AB - Sheep are one of the few animal models in which natural variations in male sexual preferences have been studied experimentally. Approximately 8% of rams exhibit sexual preferences for male partners (male-oriented rams) in contrast to most rams, which prefer female partners (female-oriented rams). We identified a cell group within the medial preoptic area/anterior hypothalamus of age-matched adult sheep that was significantly larger in adult rams than in ewes. This cell group was labeled the ovine sexually dimorphic nucleus (oSDN). In addition to a sex difference, we found that the volume of the oSDN was two times greater in female oriented rams than in male-oriented rams. The dense cluster of neurons that comprise the oSDN express cytochrome P450 aromatase. Aromatase mRNA levels in the oSDN were significantly greater in female-oriented rams than in ewes, whereas male-oriented rams exhibited intermediate levels of expression. Because the medial preoptic area/anterior hypothalamus is known to control the expression of male sexual behaviors, these results suggest that naturally occurring variations in sexual partner preferences may be related to differences in brain anatomy and capacity for estrogen synthesis. PMID- 14525916 TI - Basal and adrenocorticotropin-stimulated corticosterone in the neonatal rat exposed to hypoxia from birth: modulation by chemical sympathectomy. AB - We previously demonstrated that 7-d-old rat pups exposed to hypoxia from birth exhibit ACTH-independent increases in corticosterone associated with an increase in steroidogenic acute regulatory (StAR) and peripheral-type benzodiazepine receptor (PBR) proteins. The purpose of the present study was to determine whether this increase in corticosterone could be attenuated by chemical sympathectomy induced with guanethidine treatment. Rat pups were exposed to normoxia or hypoxia from birth and treated with vehicle or guanethidine and studied at 7 d of age. Hypoxia per se resulted in an increase in plasma corticosterone without a change in plasma ACTH. Guanethidine treatment attenuated the increase in basal corticosterone in hypoxic pups but did not attenuate ACTH stimulated corticosterone production. This effect was specific as basal and ACTH stimulated aldosterone was not affected. Guanethidine also attenuated the increase in StAR protein induced by hypoxia. Neither the effect of hypoxia nor that of guanethidine could be explained by changes in the levels of adrenal tyrosine hydroxylase, StAR, or P450scc mRNA, adrenal tyrosine hydroxylase immunohistochemistry, or adrenal catecholamine content. We conclude that chemical sympathectomy normalizes basal corticosterone levels but has no effect on ACTH stimulated corticosterone levels in 7-d-old rats exposed to hypoxia from birth. The mechanism of the effect of guanethidine to normalize hypoxia-stimulated basal corticosterone remains to be identified, although StAR protein may be an important mediator. This ACTH-independent increase in corticosterone may be a mechanism by which the neonate can increase circulating glucocorticoids necessary for survival while bypassing the hyporesponsiveness of the neonatal hypothalamic pituitary-adrenal axis. PMID- 14525917 TI - Diabetes causes decreased osteoclastogenesis, reduced bone formation, and enhanced apoptosis of osteoblastic cells in bacteria stimulated bone loss. AB - The most common cause of inflammatory bone loss is periodontal disease. After bacterial insult, inflammation induces bone resorption, which is followed by new reparative bone formation. Because diabetics have a higher incidence and more severe periodontitis, we examined mechanisms by which diabetes alters the response of bone to bacterial challenge. This was accomplished with db/db mice, which naturally develop type 2 diabetes. After inoculation of bacteria osteoclastogenesis and bone resorption was measured. Both parameters were decreased in the diabetic group. Diabetes also suppressed reparative bone formation measured histologically and by the expression of osteocalcin. The impact of diabetes on new bone formation coincided with the effect of diabetes on apoptosis of bone-lining cells. Within 5 d of bacterial challenge, apoptosis declined in the wild-type animals yet remained significantly higher in the diabetic group. Thus, diabetes may cause a net loss of bone because the suppression of bone formation is greater than the suppression of bone resorption. The uncoupling of bone formation and resorption may be due in part to prolonged apoptosis of bone lining cells. PMID- 14525918 TI - The evolution of the discipline of vascular biology: from systems physiology to molecular biology to molecular systems. PMID- 14525919 TI - Brain in the brawn: the neuronal nitric oxide synthase as a regulator of myogenic tone. PMID- 14525920 TI - A surprising new arrhythmia mechanism in heart failure. PMID- 14525921 TI - Identification of novel signaling complexes by functional proteomics. AB - The rapid development of proteomic technologies, combined with the completion of the Human Genome Map, has enabled the compiling of an unprecedented inventory of cellular proteins. Functional proteomics is an emerging field that aims to utilize the enormous amount of information provided by these proteomic technologies to understand the functions of cellular proteins. The utility of functional proteomics has been recently exploited to elucidate cellular mechanisms in numerous fields, of particular salience in the area of signal transduction. This review presents a functional proteomic approach for the study of cardiac cell signaling. It illustrates the strategies by which the subproteome of a targeted signaling system is characterized in an unbiased fashion, the manner in which the biochemical functions of this subproteome are assessed using established molecular and protein chemistry methods, and the challenges associated with these studies. PMID- 14525922 TI - The Janus faces of iNOS. PMID- 14525924 TI - Extensive exon reshuffling over evolutionary time coupled to trans-splicing in Drosophila. AB - The relative position of exons in genes can be altered only after large structural mutations. These mutations are frequently deleterious, impairing transcription, splicing, RNA stability, or protein function, as well as imposing strong inflexibility to protein evolution. Alternative cis- or trans-splicing may overcome the need for genomic structural stability, allowing genes to encode new proteins without the need to maintain a specific exon order. Trans-splicing in the Drosophila melanogaster modifier of mdg4 (mod[mdg4]) gene is the best documented example in which this process plays a major role in the maturation of mRNAs. Comparison of the genomic organization of this locus among several insect species suggests that the divergence between the lineages of the mosquito Anopheles gambiae and D. melanogaster involved an extensive exon rearrangement, requiring >11 breakpoints within the mod(mdg4) gene. The massive reorganization of the locus also included the deletion or addition of a new function as well as exon duplications. Whereas both DNA strands are sense strands in the Drosophila gene, the coding region in mosquito lays in a single strand, suggesting that trans-splicing may have originated in the Drosophila lineage and might have been the triggering factor for such a dramatic reorganization. PMID- 14525925 TI - Gene loss, protein sequence divergence, gene dispensability, expression level, and interactivity are correlated in eukaryotic evolution. AB - Lineage-specific gene loss, to a large extent, accounts for the differences in gene repertoires between genomes, particularly among eukaryotes. We derived a parsimonious scenario of gene losses for eukaryotic orthologous groups (KOGs) from seven complete eukaryotic genomes. The scenario involves substantial gene loss in fungi, nematodes, and insects. Based on this evolutionary scenario and estimates of the divergence times between major eukaryotic phyla, we introduce a numerical measure, the propensity for gene loss (PGL). We explore the connection among the propensity of a gene to be lost in evolution (PGL value), protein sequence divergence, the effect of gene knockout on fitness, the number of protein-protein interactions, and expression level for the genes in KOGs. Significant correlations between PGL and each of these variables were detected. Genes that have a lower propensity to be lost in eukaryotic evolution accumulate fewer substitutions in their protein sequences and tend to be essential for the organism viability, tend to be highly expressed, and have many interaction partners. The dependence between PGL and gene dispensability and interactivity is much stronger than that for sequence evolution rate. Thus, propensity of a gene to be lost during evolution seems to be a direct reflection of its biological importance. PMID- 14525923 TI - An evolutionary analysis of orphan genes in Drosophila. AB - Orphan genes are protein-coding regions that have no recognizable homolog in distantly related species. A substantial fraction of coding regions in any genome sequenced consists of orphan genes, but the evolutionary and functional significance of orphan genes is not understood. We present a reanalysis of the Drosophila melanogaster proteome that shows that there are still between 26% and 29% of all proteins without a significant match with noninsect sequences, and that these orphans are underrepresented in genetic screens. To analyze the characteristics of orphan genes in Drosophila, we used sequence comparisons between cDNAs retrieved from two Drosophila yakuba libraries and their corresponding D. melanogaster orthologs. We find that a cDNA library from adults yields twice as many orphan genes as such a library from embryos. The orphan genes evolve on average more than three times faster than nonorphan genes, although the width of the evolutionary rate distribution is similar for the two classes. In particular, some orphan genes show very low substitution rates that are comparable to otherwise highly conserved genes. We propose a model suggesting that orphans may be involved in the evolution of adaptive traits, and that slow evolving orphan genes may be particularly interesting candidate genes for identifying lineage-specific adaptations. PMID- 14525926 TI - Two distinct modes of microsatellite mutation processes: evidence from the complete genomic sequences of nine species. AB - We surveyed microsatellite distribution in 10 completely sequenced genomes. Using a permutation-based statistic, we assessed for all 10 genomes whether the microsatellite distribution significantly differed from expectations. Consistent with previous reports, we observed a highly significant excess of long microsatellites. Focusing on short microsatellites containing only a few repeat units, we demonstrate that this repeat class is significantly underrepresented in most genomes. This pattern was observed across different repeat types. Computer simulations indicated that neither base substitutions nor a combination of length dependent slippage and base substitutions could explain the observed pattern of microsatellite distribution. When we introduced one additional mutation process, a length-independent slippage (indel slippage) operating at repeats with few repetitions, our computer simulations captured the observed pattern of microsatellite distribution. PMID- 14525927 TI - Adaptive evolution of MRG, a neuron-specific gene family implicated in nociception. AB - The MRG gene family (also known as SNSR) belongs to the G-protein-coupled receptor (GPCR) superfamily, is expressed specifically in nociceptive neurons, and is implicated in the modulation of nociception. Here, we show that Ka/Ks (the ratio between nonsynonymous and synonymous substitution rates) displays distinct profiles along the coding regions of MRG, with peaks (Ka/Ks>1) corresponding to extracellular domains, and valleys (Ka/Ks<1) corresponding to transmembrane and cytoplasmic domains. The extracellular domains are also characterized by a significant excess of radical amino acid changes. Statistical analysis shows that positive selection is by far the most suitable model to account for the nucleotide substitution patterns in MRG. Together, these results demonstrate that the extracellular domains of the MRG receptor family, which presumably partake in ligand binding, have experienced strong positive selection. Such selection is likely directed at altering the sensitivity and/or selectivity of nociceptive neurons to aversive stimuli. Thus, our finding suggests pain perception as an aspect of the nervous system that may have experienced a surprising level of adaptive evolution. PMID- 14525928 TI - Single nucleotide variation analysis in 65 candidate genes for CNS disorders in a representative sample of the European population. AB - The detailed investigation of variation in functionally important regions of the human genome is expected to promote understanding of genetically complex diseases. We resequenced 65 candidate genes for CNS disorders in an average of 85 European individuals. The minor allele frequency (MAF), an indicator of weak purifying selection, was lowest in radical amino acid alterations, whereas similar MAF was observed for synonymous variants and conservative amino acid alterations. In noncoding sequences, variants located in CpG islands tended to have a lower MAF than those outside CpG islands. The transition/transversion ratio was increased among both synonymous and conservative variants compared with noncoding variants. Conversely, the transition/transversion ratio was lowest among radical amino acid alterations. Furthermore, among nonsynonymous variants, transversions displayed lower MAF than did transitions. This suggests that transversions are associated with functionally important amino acid alterations. By comparing our data with public SNP databases, we found that variants with lower allele frequency are underrepresented in these databases. Therefore, radical variants obtain distinctively lower database coverage. However, those variants appear to be under weak purifying selection and thus could play a role in the etiology of genetically complex diseases. PMID- 14525929 TI - Ethnic India: a genomic view, with special reference to peopling and structure. AB - We report a comprehensive statistical analysis of data on 58 DNA markers (mitochondrial [mt], Y-chromosomal, and autosomal) and sequence data of the mtHVS1 from a large number of ethnically diverse populations of India. Our results provide genomic evidence that (1) there is an underlying unity of female lineages in India, indicating that the initial number of female settlers may have been small; (2) the tribal and the caste populations are highly differentiated; (3) the Austro-Asiatic tribals are the earliest settlers in India, providing support to one anthropological hypothesis while refuting some others; (4) a major wave of humans entered India through the northeast; (5) the Tibeto-Burman tribals share considerable genetic commonalities with the Austro-Asiatic tribals, supporting the hypothesis that they may have shared a common habitat in southern China, but the two groups of tribals can be differentiated on the basis of Y chromosomal haplotypes; (6) the Dravidian tribals were possibly widespread throughout India before the arrival of the Indo-European-speaking nomads, but retreated to southern India to avoid dominance; (7) formation of populations by fission that resulted in founder and drift effects have left their imprints on the genetic structures of contemporary populations; (8) the upper castes show closer genetic affinities with Central Asian populations, although those of southern India are more distant than those of northern India; (9) historical gene flow into India has contributed to a considerable obliteration of genetic histories of contemporary populations so that there is at present no clear congruence of genetic and geographical or sociocultural affinities. PMID- 14525930 TI - DNA enrichment by allele-specific hybridization (DEASH): a novel method for haplotyping and for detecting low-frequency base substitutional variants and recombinant DNA molecules. AB - Detecting rare sequence variants in genomic DNA is central to the analysis of de novo mutation and recombination events and the detection of rare pathological mutations in mixed cell populations. Current PCR techniques suffer from noise that limits detection to variants present at a frequency of at least 10(-4)-10( 5) per cell. We now describe an alternative approach that recovers genomic DNA molecules containing a known single-nucleotide variant by hybridization selection using a biotinylated allele-specific oligonucleotide, followed by hybrid capture on streptavidin-coated paramagnetic beads and subsequent analysis by PCR. This technique of DNA enrichment by allele-specific hybridization (DEASH) is fast, effective for all tested single-nucleotide polymorphisms (SNPs), and can recover large (>10 kb) single-stranded molecules. A single round of DEASH is effective in separating haplotypes from genomic DNA and can not only readily detect and validate DNA molecules containing a single base change at a frequency of 10(-5) per cell, but can also place these changes within the context of an extended haplotype. This technique offers a new approach to the analysis of mutation and recombination, and has the potential to detect very rare de novo base substitutions. PMID- 14525931 TI - High-throughput selection of effective RNAi probes for gene silencing. AB - RNA interference (RNAi) is a process of sequence-specific posttranscriptional gene silencing mediated by double-stranded RNA. RNAi has recently emerged as a powerful genetic tool to analyze gene function in mammalian cells. The power of this method is limited however, by the uncertainty in predicting the efficacy of small interfering RNAs (siRNAs) in silencing a gene. This has imposed serious limitations not only for small-scale but also for high-throughput RNAi screening initiatives in mammalian systems. We have developed a reliable and quantitative approach for the rapid and efficient identification of the most effective siRNA against any gene. The efficacy of siRNA sequences is monitored by their ability to reduce the expression of cognate target-reporter fusions with easily quantified readouts. Finally, using micro array-based cell transfections, we demonstrate an unlimited potential of this approach in high-throughput screens for identifying effective siRNA probes for silencing genes in mammalian systems. This approach is likely to have implications in the use of RNAi as a reverse genetic tool for analyzing mammalian gene function on a genome-wide scale. PMID- 14525932 TI - RNAi microarray analysis in cultured mammalian cells. AB - RNA interference (RNAi) mediated by small interfering RNAs (siRNAs) is a powerful new tool for analyzing gene knockdown phenotypes in living mammalian cells. To facilitate large-scale, high-throughput functional genomics studies using RNAi, we have developed a microarray-based technology for highly parallel analysis. Specifically, siRNAs in a transfection matrix were first arrayed on glass slides, overlaid with a monolayer of adherent cells, incubated to allow reverse transfection, and assessed for the effects of gene silencing by digital image analysis at a single cell level. Validation experiments with HeLa cells stably expressing GFP showed spatially confined, sequence-specific, time- and dose dependent inhibition of green fluorescence for those cells growing directly on microspots containing siRNA targeting the GFP sequence. Microarray-based siRNA transfections analyzed with a custom-made quantitative image analysis system produced results that were identical to those from traditional well-based transfection, quantified by flow cytometry. Finally, to integrate experimental details, image analysis, data display, and data archiving, we developed a prototype information management system for high-throughput cell-based analyses. In summary, this RNAi microarray platform, together with ongoing efforts to develop large-scale human siRNA libraries, should facilitate genomic-scale cell based analyses of gene function. PMID- 14525933 TI - Comprehensive analysis of orthologous protein domains using the HOPS database. AB - One of the most reliable methods for protein function annotation is to transfer experimentally known functions from orthologous proteins in other organisms. Most methods for identifying orthologs operate on a subset of organisms with a completely sequenced genome, and treat proteins as single-domain units. However, it is well known that proteins are often made up of several independent domains, and there is a wealth of protein sequences from genomes that are not completely sequenced. A comprehensive set of protein domain families is found in the Pfam database. We wanted to apply orthology detection to Pfam families, but first some issues needed to be addressed. First, orthology detection becomes impractical and unreliable when too many species are included. Second, shorter domains contain less information. It is therefore important to assess the quality of the orthology assignment and avoid very short domains altogether. We present a database of orthologous protein domains in Pfam called HOPS: Hierarchical grouping of Orthologous and Paralogous Sequences. Orthology is inferred in a hierarchic system of phylogenetic subgroups using ortholog bootstrapping. To avoid the frequent errors stemming from horizontally transferred genes in bacteria, the analysis is presently limited to eukaryotic genes. The results are accessible in the graphical browser NIFAS, a Java tool originally developed for analyzing phylogenetic relations within Pfam families. The method was tested on a set of curated orthologs with experimentally verified function. In comparison to tree reconciliation with a complete species tree, our approach finds significantly more orthologs in the test set. Examples for investigating gene fusions and domain recombination using HOPS are given. PMID- 14525935 TI - In an era of scientific opportunity, are there opportunities for biomedical scientists? AB - Has the dramatic growth in the NIH budget affected the training and production of biomedical Ph.D.s? Examination of new survey data reveals surprising findings. Despite the need for an increased workforce to carry out the expansion in biomedical research, there has not been an increase in new U.S. doctorates awarded, and time-to-degree in the biomedical sciences is no longer increasing. Furthermore, both the frequency and length of postdoctoral appointments are decreasing for U.S. biomedical science recipients. There has been, however, continued growth in the number of foreign postdoctorals. Industrial employment of biomedical scientists continues to increase, but there has been only modest growth in tenured or tenure-track academic jobs. PMID- 14525936 TI - Uncoupling protein 3 as a mitochondrial fatty acid anion exporter. AB - In contrast to UCP1, the primary function of UCP3 is not the dissipation of energy. Rather, several lines of evidence suggest that UCP3 is related to cellular long-chain fatty acid homeostasis. If long-chain fatty acids enter the mitochondrial matrix in their non-esterified form, they cannot be metabolized and may exert deleterious effects. To test the feasibility that UCP3 exports fatty acid anions, we systematically interfered at distinct steps in the fatty acid metabolism pathway, thereby creating conditions in which the entry of (non esterified) fatty acids into the mitochondrial matrix is enhanced. First, reducing the cellular fatty acid binding capacity, known to increase cytosolic concentrations of non-esterified fatty acids, up-regulated UCP3 5.3-fold. Second, inhibition of mitochondrial entry of esterified long-chain fatty acids up regulated UCP3 by 1.9-fold. Third, high-fat diets, to increase mitochondrial supply of non-esterified long-chain fatty acids exceeding oxidative capacity, up regulated UCP3 twofold. However, feeding a similar amount of medium-chain fatty acids, which can be oxidized inside the mitochondrial matrix and therefore do not need to be exported from the matrix, did not affect UCP3 protein levels. These data are compatible with a physiological function of UCP3 in facilitating outward transport of long-chain fatty acid anions, which cannot be oxidized, from the mitochondrial matrix. PMID- 14525934 TI - Development of human protein reference database as an initial platform for approaching systems biology in humans. AB - Human Protein Reference Database (HPRD) is an object database that integrates a wealth of information relevant to the function of human proteins in health and disease. Data pertaining to thousands of protein-protein interactions, posttranslational modifications, enzyme/substrate relationships, disease associations, tissue expression, and subcellular localization were extracted from the literature for a nonredundant set of 2750 human proteins. Almost all the information was obtained manually by biologists who read and interpreted >300,000 published articles during the annotation process. This database, which has an intuitive query interface allowing easy access to all the features of proteins, was built by using open source technologies and will be freely available at http://www.hprd.org to the academic community. This unified bioinformatics platform will be useful in cataloging and mining the large number of proteomic interactions and alterations that will be discovered in the postgenomic era. PMID- 14525937 TI - Nitric oxide inhibits the adenovirus proteinase in vitro and viral infectivity in vivo. AB - Nitric oxide (NO) is an antiviral effector of the innate immune system, but few of the viral targets of NO have been identified. We now show that NO inhibits adenovirus replication by targeting the adenovirus proteinase (AVP). NO generated from diethylamine NONOate (DEA-NONOate) or spermine NONOate (Sp-NONOate) inhibited the AVP. Inhibition was reversible with dithiothreitol. The equilibrium dissociation constant for reversible binding to the AVP by Sp-NONOate, or Ki, was 0.47 mM, and the first-order rate constant for irreversible inhibition of the AVP by Sp-NONOate, or ki, was 0.0036 s(-1). Two hallmarks of a successful adenovirus infection were abolished by the NO donors: the appearance of E1A protein and the cleavage of cytokeratin 18 by AVP. Treatment of infectious virus by DEA-NONOate dramatically decreased viral infectivity. These data suggest that NO may be a useful antiviral agent against viruses encoding a cysteine proteinase and in particular may be an antiadenovirus agent. PMID- 14525938 TI - Expression of the small heat-shock protein Hsp16-2 in Caenorhabditis elegans is suppressed by Ginkgo biloba extract EGb 761. AB - EGb 761, a standardized extract of Ginkgo biloba leaves, has been shown to have antioxidative properties. We have previously demonstrated that EGb 761 increases stress resistance and mean life span in the model organism Caenorhabditis elegans. In this study, the molecular mechanism of EGb 761 on alleviating effects of oxidative stress is further investigated using transgenic C. elegans expressing a jellyfish green fluorescent protein (GFP)-tagged inducible small heat-shock protein gene (hsp-16-2). The expression of hsp-16-2 induced by the pro oxidant juglone and by heat shock was significantly suppressed by 86% and 33%, respectively, in the transgenic nematode fed with EGb 761. These effects of EGb 761 correlate with its ability to increase mean survival rate of the nematode in response to acute oxidative and thermal stresses, as well as to attenuate the basal levels of hydrogen peroxide in the organism. Thus, we interpret the suppression of hsp-16-2/GFP expression as an indication that EGb 761 decreases cellular stress resulting from exogenous treatments, therefore leading to a decreased transcriptional induction of the reporter transgene. These results support the hypothesis that EGb 761 augments the natural antistress system of C. elegans, thus increasing stress resistance and life span. PMID- 14525940 TI - Disruption of vascular endothelial homeostasis by tobacco smoke: impact on atherosclerosis. AB - The World Health Organization (WHO) predicts that by 2020 tobacco will become the largest single health problem worldwide and will cause an estimated 8.4 million deaths annually (http://www5.who.int/tobacco/). Although the impact of smoking on human health is well defined from the medical point of view, surprisingly little is known about the mechanisms by which tobacco smoke mediates its disastrous effects. Here, we demonstrate that tobacco smoke dramatically changes vascular endothelial cell and tissue morphology, leading to a loss of endothelial barrier function within minutes. Long-term exposure of endothelial cells to tobacco smoke extracts induces necrosis that may trigger a pro-inflammatory status of the vessel wall. Pre-incubation of the extracts without cells for 6 h at 37 degrees C led to a complete loss of activity. Further, the endothelium could be rescued by changing to fresh medium even at times when the extracts had lost their activity. Finally, we show that N-acetyl cysteine and statins inhibit the adverse tobacco smoke effects. PMID- 14525939 TI - The presence of nitrite during UVA irradiation protects from apoptosis. AB - Nitrite occurs ubiquitously in biological fluids such as blood and sweat, representing an oxidation product of nitric oxide. Nitrite has been associated with a variety of adverse effects such as mutagenicity, carcinogenesis, and toxicity. In contrast, here we demonstrate that the presence of nitrite, but not nitrate, during irradiation of endothelial cells in culture exerts a potent and concentration-dependent protection against UVA-induced apoptotic cell death. Protection is half-maximal at a concentration of 3 mM, and complete rescue is observed at 10 mM. Nitrite-mediated protection is mediated via inhibition of lipid peroxidation in a similar manner as seen with butylated hydroxytoluene, a known inhibitor of lipid peroxidation. Interestingly, nitrite-mediated protection is completely abolished by coincubation with the NO scavenger cPTIO. Using electron paramagnetic resonance (EPR) spectroscopy or Faraday modulation spectroscopy, we directly prove UVA-induced NO formation in solutions containing nitrite. In conclusion, evidence is presented that nitrite represents a protective agent against UVA-induced apoptosis due to photodecomposition of nitrite and subsequent formation of NO. PMID- 14525941 TI - Sonic hedgehog is a neuromodulator in the adult subthalamic nucleus. AB - It is well established that members of the hedgehog family are involved in tissue patterning during development. We herein show that sonic hedgehog signaling molecules are differentially regulated by dopamine depletion in the basal ganglia of adult animals and specifically that sonic hedgehog levels are reduced in an animal model of Parkinson's disease. In addition, we show that sonic hedgehog protein inhibits electrical activity in the subthalamic nucleus, a key element of basal ganglia, within minutes of application. As the subthalamic nucleus is overactive in parkinsonism, we suggest that enhancement of sonic hedgehog signaling in the subthalamic nucleus may be of therapeutic value in Parkinson's disease. PMID- 14525942 TI - Peroxisome proliferator-activated receptor delta controls muscle development and oxidative capability. AB - Peroxisome proliferator-activated receptors (PPARs) are nuclear receptors exerting several functions in development and metabolism. The physiological functions of PPARdelta remain elusive. By using a CRE-Lox recombination approach, we generated an animal model for muscle-specific PPARdelta overexpression to investigate the role of PPARdelta in this tissue. Muscle-specific PPARdelta overexpression results in a profound change in fiber composition due to hyperplasia and/or shift to more oxidative fiber and, as a consequence, leads to the increase of both enzymatic activities and genes implicated in oxidative metabolism. These changes in muscle are accompanied by a reduction of body fat mass, mainly due to a large reduction of adipose cell size. Furthermore, we demonstrate that endurance exercise promotes an accumulation of PPARdelta protein in muscle of wild-type animals. Collectively, these results suggest that PPARdelta plays an important role in muscle development and adaptive response to environmental changes, such as training exercise. They strongly support the idea that activation of PPARdelta could be beneficial in prevention of metabolic disorders, such as obesity or type 2 diabetes. PMID- 14525943 TI - Sorbitol dehydrogenase: a novel target for adjunctive protection of ischemic myocardium. AB - Sorbitol dehydrogenase (SDH) is a polyol pathway enzyme that catalyzes conversion of sorbitol to fructose. Recent studies have demonstrated that activation of aldose reductase, the first enzyme of the polyol pathway, is a key response to ischemia and that inhibition of aldose reductase reduces myocardial ischemic injury. In our efforts to understand the role of pathway in affecting metabolism under normoxic and ischemic conditions, as well as in ischemic injury in myocardium, we investigated the importance of SDH by use of a specific inhibitor (SDI), CP-470,711. SDH inhibition increased glucose oxidation, whereas palmitate oxidation remained unaffected. Global ischemia increased myocardial SDH activity by approximately 1.5 fold. The tissue lactate/pyruvate ratio, a measure of cytosolic NADH/NAD+, was reduced by SDH inhibition under both normoxic and ischemic conditions. ATP was higher in SDI hearts during ischemia and reperfusion. Creatine kinase release during reperfusion, a marker of myocardial ischemic injury, was markedly attenuated in SDH-inhibited hearts. These data indicate that myocardial SDH activation is a component of ischemic response and that interventions that inhibit SDH protect ischemic myocardium. Furthermore, these data identify SDH as a novel target for adjunctive cardioprotective interventions. PMID- 14525944 TI - The importance of propargylamine moiety in the anti-Parkinson drug rasagiline and its derivatives in MAPK-dependent amyloid precursor protein processing. AB - Rasagiline [N-propargyl-(1R)-aminoindan] a highly potent selective irreversible monoamine oxidase (MAO)-B inhibitor exerts neuroprotective and antiapoptotic effects against a variety of insults in cell cultures and in vivo and has finished its phase III clinical trials for Parkinson's disease. In the present study, we show that rasagiline (1 and 10 microM) significantly protected rat PC12 cells against beta-amyloid (Abeta1-42) toxicity. In addition, rasagiline significantly increased (approximately threefold) the secretion of the nonamyloidogenic soluble form of the amyloid precursor protein (sAPPalpha) from SH-SY5Y neuroblastoma and PC12 cells. The increase of sAPPalpha was dose dependent and was blocked by the hydroxamic acid-based metalloprotease inhibitor Ro31-9790 (100 microM), suggesting that the effect is mediated via alpha secretase activity. Rasagiline-induced sAPPalpha release was significantly reduced by the inhibitors of protein kinase C (PKC), GF109203X, and ERK mitogen activated protein kinase (MAPK) PD98059. Moreover, rasagiline dose dependently (0.1-10 microM) increased the phosphorylation of p44 and p42 MAPK, which was abolished by PD98059 (30 microM) and GF109203X (2.5 microM). By comparing the actions of rasagiline with those of its S-isomer TVP1022, which is not an MAO inhibitor, we have been able to demonstrate that MAO-B inhibition is not a prerequisite for either sAPPalpha-induced release or ERK phosphorylation. In addition, structure-activity relationship among rasagiline-related compounds suggests the crucial role of the propargyl moiety in these molecules, because propargylamine itself significantly induced the secretion of sAPPalpha and increased MAPK phosphorylation with similar potency to that of rasagiline and its derivatives. PMID- 14525945 TI - Divergent effects of GM-CSF and TGFbeta1 on bone marrow-derived macrophage arginase-1 activity, MCP-1 expression, and matrix metalloproteinase-12: a potential role during arteriogenesis. AB - Granulocyte/macrophage-colony stimulating factor (GM-CSF) and transforming growth factor (TGF)beta1 induce arteriogenesis in a nonischemic model of femoral artery ligation. Moreover, clinical trials demonstrated an improved collateralization after injection of bone marrow cells. In the present study, the expression of arteriogenic factors in bone marrow-derived macrophages (BMDM) was measured to verify the potential of these cells to influence collateral artery growth. GM-CSF induced in BMDM the expression of monocyte chemoattractive protein (MCP)-1, matrix-metalloproteinase (MMP)-12, and arginase-1-the latter also showing a remarkable increase in activity. During in vivo induced arteriogenesis, the accumulation rate of macrophages around proliferating collaterals was significantly increased. We also show that MCP-1 is found to be mainly expressed in the media of the vessel wall, MMP-12 in macrophages of the adventitia, and arginase at both locations. This study provides for the first time a comprehensive analysis of GM-CSF/TGFbeta1-regulated arteriogenic factors in BMDM and supports the hypothesis that arteriogenesis is a multistage mechanism, including monocyte/macrophage adhesion and transmigration, pro-arteriogenic cytokine expression, degradation of connective tissue, and collagen synthesis regulation. Selective modulation of these mechanisms as well as cell-based therapies supplying arteriogenic factors in vivo point toward new strategies to influence collateral artery growth. PMID- 14525946 TI - Coordinated functions of Akt/PKB and ETS1 in tubule formation. AB - We investigated the inter-relationship between two downstream effectors of vascular endothelial growth factor (VEGF), the serine threonine kinase Akt (also known as protein kinase B) and the transcription factor ETS1, during tubulogenesis. Human endothelial cell culture and the in vivo Drosophila tracheal systems are employed in comparative analysis. We show that VEGF stimulates the expression of ETS1 through a phosphatidylinositol-3-kinase (PI3K)/Akt-dependent pathway in primary endothelial cells. Activation of Akt results in vessel formation in vitro, a process that is blocked by expression of antisense ETS1. The functional relationship between ETS and Akt was then tested in the homologous tubular system in Drosophila. Contrary to expectation, ETS1 and Akt did not form a linear positive regulatory pathway in vivo. Instead, genetic analyses suggest that the Drosophila ETS1 homologue Pointed is required for cell motility per se while Drosophila Akt (Dakt1) is responsible for organized and restricted cell movement that is essential for tubule formation. Taken together, our results show that ETS1 and Akt control different aspects of cell motility that are integrated in the precise regulation of vascular tubule formation. PMID- 14525947 TI - Increases in weight of growth hormone-deficient and immunodeficient (lit/scid) dwarf mice after grafting of hGH-secreting, primary human keratinocytes. AB - Primary human keratinocytes, stably transduced with the human growth hormone (hGH) gene (under control of the retroviral LTR promoter) and selected via geneticin secreted as much as 7 microg hGH/106 cells/day. Their grafting onto immunodeficient dwarf mice (lit/scid) led to hGH levels in the circulation that did not go below 0.2-0.3 ng/ml during a 12 day period (peak value, 1.5 ng/ml at 4 h). This phenomenon was associated with a body weight increase of the grafted animals (0.060 g/animal/day) significantly higher (P<0.01) than that of controls (0.023 g/animal/day). This is the first report describing successful utilization of immunodeficient dwarf mice (lit/scid) in keratinocyte-based hGH gene therapy. PMID- 14525948 TI - Abnormal PTEN expression in portal hypertensive gastric mucosa: a key to impaired PI 3-kinase/Akt activation and delayed injury healing? AB - Phosphatase and tensin homologue deleted on chromosome ten (PTEN) is a dual specificity phosphatase that has activity toward both phosphorylated peptides and phospholipids. PTEN inhibits activation of Akt, the downstream effector of PI 3 kinase, which is integral to cell proliferation, migration, survival, and angiogenesis essential for tissue injury healing. PTEN expression and activation during injury healing remain unexplored. Portal hypertensive (PHT) gastric mucosa has impaired injury healing, but the underlying mechanisms remain unknown. We investigated whether impaired healing of injured PHT gastric mucosa is due to abnormal PTEN expression/activation that leads to decreased Akt activation. We also investigated the possible involvement of Egr-1, which regulates PTEN in some cells (e.g., fetal kidney epithelial cells), and TNF-alpha, which can induce Egr 1 expression. In PHT gastric mucosa 6 h after injury, PTEN protein levels were increased by 2.7-fold; unphosphorylated PTEN (reflecting activated PTEN) was increased by 2.4-fold; Akt phosphorylation (reflecting Akt activation) was reduced by 2-fold; and Egr-1 expression was increased by 3.3-fold vs. normal gastric mucosa. TNF-alpha neutralization reversed all of the above abnormalities in PHT gastric mucosa, reduced mucosal injury, and enhanced healing. We conclude that, in injured PHT gastric mucosa, overexpressed/activated PTEN leads to the reduced activation of the PI 3-kinase/Akt pathway that results in impaired injury healing. PMID- 14525949 TI - The IKr drug response is modulated by KCR1 in transfected cardiac and noncardiac cell lines. AB - The cardiac potassium channel encoded by the human ether-a-go-go related gene (HERG) is blocked by a diverse array of common therapeutic compounds. Even transient exposure to such agents may provoke the life-threatening cardiac arrhythmia torsades de pointes in some, but not all, individuals. Although the molecular and genetic factors predicting such wide variability in drug response remain unclear, known sequence variations within the coding region of HERG do not explain the adverse drug response in many cases. Although other proteins can modulate HERG function, no studies have identified protein partners capable of limiting the pharmacological sensitivity of HERG. Here we show that KCR1, a protein identified previously in rat cerebellum, is a plasma membrane-associated protein expressed at the RNA level in the human heart and can be immunoprecipitated with HERG. Functionally, KCR1 reduces the sensitivity of HERG to classic proarrhythmic HERG blockers (sotalol, quinidine, dofetilide) in both cardiac and noncardiac cell lines. We propose that KCR1, when coupled to HERG, may limit the sensitivity of HERG to proarrhythmic drug blockade and may be a rational target for modifying the proarrhythmic effects of otherwise clinically useful compounds. PMID- 14525950 TI - Lack of type XVIII collagen results in anterior ocular defects. AB - Mice lacking type XVIII collagen have defects in the posterior part of the eye, including delayed regression of the hyaloid vasculature and poor outgrowth of the retinal vessels. We report here that these mice also have a fragile iris and develop atrophy of the ciliary body. The irises of Col18a1-/- mice can be seen to adhere to the lens and cornea. After the pupils begin to function, the double layer of epithelial cells separates at the apical cell contacts, leading to defoliation of its posterior pigment epithelial cell layer, and extracellular material begins to accumulate in the basement membrane zones of the iris. In contrast to the iris epithelia, where no clear signs of cellular atrophy were detected, the lack of type XVIII collagen resulted in atrophy of the pigmented epithelial cells of the ciliary body, and there were also ultrastructural abnormalities in the basement membrane zones. These changes did not lead to chronically elevated intraocular pressures, however. Our results indicate that type XVIII collagen is needed for the integrity of the epithelial basement membranes of the iris and the ciliary body and that its gene should therefore be taken into account as a new potential cause of anterior segment disorders in the eye. PMID- 14525951 TI - Akt/PTEN signaling mediates estrogen-dependent proliferation of primordial germ cells in vitro. AB - Testicular tumors in humans are reported to be significantly increasing in incidence. Embryo exposure to environmental estrogens has been proposed as one of the possible underlying causes. In mice, genetic, immunological, and experimental evidence suggest that germ cell testicular tumors may derive from primordial germ cells (PGCs), the embryonic precursors of gametes. Here we show that relatively high concentrations of estrogens stimulate mouse PGC growth in vitro through the somatic cells of the gonadal ridges. Moreover, we found that estrogens stimulate the transcription of the Steel gene and the production of c-Kit ligand in gonadal somatic cells, and that this growth factor is likely to be responsible for the observed stimulation of PGC growth via an Akt/PTEN pathway. Finally, we show that estrogen stimulation of gonadal somatic cells in culture, in combination with PTEN down-regulation in PGCs and the presence of leukemia inhibitory factor in the culture medium, result in high frequency of PGC transformation in tumorigenic cells. Based on these results, we present a novel experimental in vitro model for tumorigenic germ cell transformation and identify molecular pathways likely involved in development of germ cell tumors after estrogen exposure. PMID- 14525952 TI - Functional analysis of leptin receptor activation using a Janus kinase/signal transducer and activator of transcription complementation assay. AB - The leptin receptor (LR), a member of the class I cytokine receptor family, is composed of a single subunit. Its extracellular domain consists of two so-called cytokine receptor homology domains, separated by an Ig-like domain, and two additional fibronectin type III modules. Requirements for LR activation were examined using a complementation strategy. Two LR mutants, LR-FFY-Deltabox 1 and LR-F3, deficient in Janus kinase or signal transducer and activator of transcription (STAT) activation, respectively, were only able to generate a STAT3 dependent signal when coexpressed. Based on the requirements for Janus kinase/STAT signaling, and on the lack of complementation with similar receptor constructs, but containing the extracellular domain of the homodimeric erythropoietin receptor, this observation can be explained only by higher order LR clustering. Using a panel of deletion mutants we were able to define a role for the cytokine receptor homology 1 and Ig-like domains in leptin signaling. Moreover, we demonstrate a nonredundant function for the individual receptor chains within the homomeric LR complex. Based on these data, we propose a possible model for LR clustering. PMID- 14525953 TI - Position of Pro and Ser near Glu7.32 in the extracellular loop 3 of mammalian and nonmammalian gonadotropin-releasing hormone (GnRH) receptors is a critical determinant for differential ligand selectivity for mammalian GnRH and chicken GnRH-II. AB - A Glu/Asp7.32 residue in the extracellular loop 3 of the mammalian GnRH receptor (GnRHR) is known to interact with Arg8 of mammalian GnRH (mGnRH), which may confer preferential ligand selectivity for mGnRH than for chicken GnRH-II (cGnRH II). However, some nonmammalian GnRHRs also have the Glu/Asp residue at the same position, yet respond better to cGnRH-II than mGnRH. Amino acids flanking Glu/Asp7.32 are differentially arranged such that mammalian and nonmammalian GnRHRs have an S-E/D-P motif and P-X-S/Y motif, respectively. We presumed the position of Ser7.31 or Pro7.33 of rat GnRHR as a potential determinant for ligand selectivity. Either placing Pro before Glu7.32 or placing Ser after Glu7.32 significantly decreased the sensitivity and/or efficacy for mGnRH, but slightly increased that for cGnRH-II in several mutant receptors. Among them, those with a PEV, PES, or SES motif exhibited a marked decrease in sensitivity for mGnRH such that cGnRH-II had a higher potency than mGnRH, showing a reversed preferential ligand selectivity. Chimeric mGnRHs in which positions 5, 7, and/or 8 were replaced by those of cGnRH-II revealed a greater ability to activate these mutant receptors than mGnRH, whereas they were less potent to activate wild-type rat GnRHR than mGnRH. Interestingly, a mutant bullfrog type I receptor with the SEP motif exhibited an increased sensitivity for mGnRH but a decreased sensitivity for cGnRH-II. These results indicate that the position of Pro and Ser near Glu7.32 in the extracellular loop 3 is critical for the differential ligand selectivity between mammalian and nonmammalian GnRHRs. PMID- 14525954 TI - The peroxisome proliferator-activated receptor beta/delta agonist, GW501516, regulates the expression of genes involved in lipid catabolism and energy uncoupling in skeletal muscle cells. AB - Lipid homeostasis is controlled by the peroxisome proliferator-activated receptors (PPARalpha, -beta/delta, and -gamma) that function as fatty acid dependent DNA-binding proteins that regulate lipid metabolism. In vitro and in vivo genetic and pharmacological studies have demonstrated PPARalpha regulates lipid catabolism. In contrast, PPARgamma regulates the conflicting process of lipid storage. However, relatively little is known about PPARbeta/delta in the context of target tissues, target genes, lipid homeostasis, and functional overlap with PPARalpha and -gamma. PPARbeta/delta, a very low-density lipoprotein sensor, is abundantly expressed in skeletal muscle, a major mass peripheral tissue that accounts for approximately 40% of total body weight. Skeletal muscle is a metabolically active tissue, and a primary site of glucose metabolism, fatty acid oxidation, and cholesterol efflux. Consequently, it has a significant role in insulin sensitivity, the blood-lipid profile, and lipid homeostasis. Surprisingly, the role of PPARbeta/delta in skeletal muscle has not been investigated. We utilize selective PPARalpha, -beta/delta, -gamma, and liver X receptor agonists in skeletal muscle cells to understand the functional role of PPARbeta/delta, and the complementary and/or contrasting roles of PPARs in this major mass peripheral tissue. Activation of PPARbeta/delta by GW501516 in skeletal muscle cells induces the expression of genes involved in preferential lipid utilization, beta-oxidation, cholesterol efflux, and energy uncoupling. Furthermore, we show that treatment of muscle cells with GW501516 increases apolipoprotein-A1 specific efflux of intracellular cholesterol, thus identifying this tissue as an important target of PPARbeta/delta agonists. Interestingly, fenofibrate induces genes involved in fructose uptake, and glycogen formation. In contrast, rosiglitazone-mediated activation of PPARgamma induces gene expression associated with glucose uptake, fatty acid synthesis, and lipid storage. Furthermore, we show that the PPAR-dependent reporter in the muscle carnitine palmitoyl-transferase-1 promoter is directly regulated by PPARbeta/delta, and not PPARalpha in skeletal muscle cells in a PPARgamma coactivator-1-dependent manner. This study demonstrates that PPARs have distinct roles in skeletal muscle cells with respect to the regulation of lipid, carbohydrate, and energy homeostasis. Moreover, we surmise that PPARbeta/delta agonists would increase fatty acid catabolism, cholesterol efflux, and energy expenditure in muscle, and speculate selective activators of PPARbeta/delta may have therapeutic utility in the treatment of hyperlipidemia, atherosclerosis, and obesity. PMID- 14525955 TI - Nerve growth factor restores p53 function in pituitary tumor cell lines via trkA mediated activation of phosphatidylinositol 3-kinase. AB - Two groups of prolactinomas were identified, one slowly proliferating and responsive to bromocriptine and one fast proliferating and bromocriptine resistant. Nerve growth factor (NGF) inhibits proliferation of bromocriptine resistant cells by mechanisms that are still unclear. The tumor suppressor p53 is one of the key regulators of cell proliferation and in most tumors, but not pituitary adenomas, it is inactivated by genomic mutations. Here we investigated whether in prolactinoma cell lines NGF influences cell cycle-related pathways involving p53. By using conformation-specific antibodies and immunocytochemistry we found that in bromocriptine-resistant cells p53 adopts a mutant conformation that precludes its nuclear translocation and transcriptional activity. NGF administration to these cells refolds p53 into wild-type tertiary structure, promotes its nuclear translocation, and restores its DNA-binding activity as demonstrated by the transcriptional activation of p21Cip1/WAF1 and the resulting down-regulation of different cyclins and cyclin-dependent kinase 2. Inactivation of trkA, but not of p75NTR, and wortmannin prevented NGF-induced p53 nuclear translocation. Thus, in prolactinoma cells p53 is inactivated by conformational mutation and cytoplasmic segregation. This defect is reversible because NGF reconstitutes active p53 in these cells. This effect of NGF is exclusively mediated by trkA through activation of phosphatidylinositol-3-kinase and may be related to its growth-inhibitory action. PMID- 14525956 TI - Comparative genomic analysis of the eight-membered ring cystine knot-containing bone morphogenetic protein antagonists. AB - TGF-beta family proteins with a cystine knot motif serve as ligands for diverse families of plasma membrane receptors. Bone morphogenetic protein (BMP) antagonists represent a subgroup of these proteins, some of which bind BMPs and antagonize their actions during development and morphogenesis. Availability of completed genome sequences from diverse organisms allows bioinformatic analysis of the evolution of BMP antagonists and facilitates their classification. Using a regular expression algorithm (http://BioRegEx.stanford.edu), an exhaustive search of the human genome identified all cystine knot-containing BMP antagonists. Based on the size of the cystine ring, these proteins were divided into three subfamilies: CAN (eight-membered ring), twisted gastrulation (nine-membered ring), as well as chordin and noggin (10-membered ring). The CAN family can be divided further into four subgroups based on a conserved arrangement of additional cysteine residues-gremlin and PRDC, cerberus and coco, and DAN, together with USAG-1 and sclerostin. We searched for orthologs of human BMP antagonists in the genomes of model organisms and analyzed their phylogenetic relationship. New human paralogs were identified together with the verification of orthologous relationships of known genes. We also discuss the physiological roles of the CAN subfamily of BMP antagonists and the associated genetic defects. Based on the known three-dimensional structure of key cystine knot proteins, we postulated disulfide bondings for eight-membered ring BMP antagonists to predict their potential folding and dimerization. PMID- 14525957 TI - Structural determinants for vitamin D receptor response to endocrine and xenobiotic signals. AB - The vitamin D receptor (VDR), initially identified as a nuclear receptor for 1alpha,25-dihydroxyvitamin D3 [1alpha,25(OH)2D3], regulates calcium metabolism, cellular proliferation and differentiation, immune responses, and other physiological processes. Recently, secondary bile acids such as lithocholic acid (LCA) were identified as endogenous VDR agonists. To identify structural determinants required for VDR activation by 1alpha,25(OH)2D3 and LCA, we generated VDR mutants predicted to modulate ligand response based on sequence homology to pregnane X receptor, another bile acid-responsive nuclear receptor. In both vitamin D response element activation and mammalian two-hybrid assays, we found that VDR-S278V is activated by 1alpha,25(OH)2D3 but not by LCA, whereas VDR S237M can respond to LCA but not to 1alpha,25(OH)2D3. Competitive ligand binding analysis reveals that LCA, but not 1alpha,25(OH)2D3, effectively binds to VDR S237M and both 1alpha,25(OH)2D3 and LCA bind to VDR-S278V. We propose a docking model for LCA binding to VDR that is supported by mutagenesis data. Comparative analysis of the VDR-LCA and VDR-1alpha,25(OH)2D3 structure-activity relationships should be useful in the development of bile acid-derived synthetic VDR ligands that selectively target VDR function in cancer and immune disorders without inducing adverse hypercalcemic effects. PMID- 14525958 TI - Development of Protacs to target cancer-promoting proteins for ubiquitination and degradation. AB - The proteome contains hundreds of proteins that in theory could be excellent therapeutic targets for the treatment of human diseases. However, many of these proteins are from functional classes that have never been validated as viable candidates for the development of small molecule inhibitors. Thus, to exploit fully the potential of the Human Genome Project to advance human medicine, there is a need to develop generic methods of inhibiting protein activity that do not rely on the target protein's function. We previously demonstrated that a normally stable protein, methionine aminopeptidase-2 or MetAP-2, could be artificially targeted to an Skp1-Cullin-F-box (SCF) ubiquitin ligase complex for ubiquitination and degradation through a chimeric bridging molecule or Protac (proteolysis targeting chimeric molecule). This Protac consisted of an SCF(beta TRCP)-binding phosphopeptide derived from IkappaBalpha linked to ovalicin, which covalently binds MetAP-2. In this study, we employed this approach to target two different proteins, the estrogen (ER) and androgen (AR) receptors, which have been implicated in the progression of breast and prostate cancer, respectively. We show here that an estradiol-based Protac can enforce the ubiquitination and degradation of the alpha isoform of ER in vitro, and a dihydroxytestosterone based Protac introduced into cells promotes the rapid disappearance of AR in a proteasome-dependent manner. Future improvements to this technology may yield a general approach to treat a number of human diseases, including cancer. PMID- 14525960 TI - Notch signaling in the immune system. AB - Notch signaling plays a preeminent role during development in not only regulating cell fate decisions, but it can also influence growth and survival of progenitor cells. In the immune system, Notch is required for the maintenance of hematopoietic stem cells and in directing T- versus B-lineage commitment. In this review, I will summarize some of the recent findings relating to the function of Notch in the immune system during lymphocyte development and in the generation and function of mature cells. PMID- 14525959 TI - Apoptosis-induced proteinase 3 membrane expression is independent from degranulation. AB - Proteinase 3 (PR3) and human neutrophil elastase (HNE) are serine proteinases stored in the azurophilic granules of neutrophils. In contrast to HNE, PR3 is the target of antineutrophil cytoplasm antibodies (ANCA) in Wegener's granulomatosis. The mechanisms leading to the membrane expression of PR3 and HNE are still unclear and appear to be critical to understand the pathophysiological role of ANCA. Stably transfected rat basophilic cell lines (RBL) with PR3 or HNE were used to analyze the PR3 and HNE secretion mechanisms and differentiate between them. RBL cells were lacking endogenous PR3 and HNE. They were stably transfected with HNE or PR3 or an inactive mutant of PR3 (PR3S203A). Using the calcium ionophore A23187 as a secretagogue, higher serine proteinase activity was secreted in the supernatant of RBL/HNE than in RBL/PR3. It is interesting that PR3 and PR3/S203A were also expressed at the plasma membrane, thus demonstrating that serine protease activity was not required for plasma membrane expression. In contrast, no expression of plasma membrane HNE could be detected in RBL/HNE. Apoptosis induced by etoposide was evaluated by DNA fragmentation, the presence of cytoplasmic histone-associated DNA fragments, and annexin V labeling. No membrane HNE was detected in RBL/HNE. In contrast, in RBL/PR3 and in RBL/PR3S203A, the membrane expression of PR3 and PR3S203A increased with etoposide concentrations and appeared closely related to annexin V labeling. Our data suggest that membrane PR3 originates from two distinct pools, the granular pool mobilized following degranulation or a plasma membrane pool mobilized upon apoptosis. PMID- 14525961 TI - Monoclonal LYM-1 antibody-dependent cytolysis by human neutrophils exposed to GM CSF: auto-regulation of target cell attack by cathepsin G. AB - Murine monoclonal antibody (mAb) Lym-1 is an immunoglobulin G2a specific for certain human leukocyte antigen-DR variants expressed on the surface of malignant B cells. It has been proposed for serotherapy in patients with B lymphomas. We have previously shown that mAb Lym-1 synergizes with granulocyte macrophage colony stimulating factor to promote Raji B-lymphoid cell lysis by human neutrophils via the intervention of neutrophil Fc receptors type II and D-mannose inhibitable interactions between CD11b-CD18 integrins and CD66b glycoproteins. Here, we provide evidence that the process is oxygen-independent by inference related to the release of primary granules and is regulated by cathepsin G activity. The lysis was indeed reproduced by replacing normal neutrophils with cells from three patients suffering from chronic granulomatous disease, i.e., neutrophils genetically incapable of generating oxidants. Moreover, the lysis was inhibited by the serine protease inhibitor 3,4-dichloroisocoumarin and by Z glycyl-leucyl-phenyl-chloromethyl ketone (Z-Gly-Leu-Phe-CMK), which blocks cathepsin G. Conversely, the lysis was unaffected by N-methoxysuccinyl-alanyl alanyl-prolyl-alanyl-CMK (MeOSuc-Ala-Ala-Pro-Ala-CMK; elastase inhibitor) and MeOSuc-Ala-Ala-Pro-valine (Val)-CMK, which inhibits elastase and proteinase 3. The ability of neutrophils, engaged in cytolysis, to release cathepsin G was proved by detecting this enzymatic activity spectrophotometrically and immunocytochemically. Moreover, inhibition of cathepsin G activity by concentrations of Z-Gly-Leu-Phe-CMK, incapable of affecting elastase activity, was found to reduce the release of elastase and myeloperoxidase from neutrophils under conditions similar to those used for cytolytic assays. These findings suggest that neutrophils auto-regulate their lytic efficiency by controlling the exocytosis of primary granules via their cathepsin G activity. PMID- 14525962 TI - CCL16/LEC powerfully triggers effector and antigen-presenting functions of macrophages and enhances T cell cytotoxicity. AB - The human CC chemokine CCL16, a liver-expressed chemokine, enhances the killing activity of mouse peritoneal macrophages by triggering their expression of tumor necrosis factor alpha (TNF-alpha) and Fas ligand. Macrophages also respond to CCL16 by enhancing their production of monocyte chemoattractant protein-1, regulated on activation, normal T cells expressed and secreted chemokines, and interleukin (IL)-1 beta, TNF-alpha, and IL-12. The effect of CCL16 is almost as strong as that of lipopolysaccharide and interferon-gamma, two of the best macrophage activators. Moreover, CCL16-activated macrophages overexpress membrane CD80, CD86, and CD40 costimulatory molecules and extensively phagocytose tumor cell debris. On exposure to such debris, they activate a strong, tumor-specific, cytolytic response in virgin T cells. Furthermore, cytolytic T cells generated in the presence of CCL16 display a higher cytotoxicity and activate caspase-8 in tumor target cells. This ability to activate caspase-8 depends on their overexpression of TNF-alpha and Fas ligand induced by CCL16. These data reveal a new function for CCL16 in the immune-response scenario. CCL16 significantly enhances the effector and the antigen-presenting function of macrophages and augments T cell lytic activity. PMID- 14525963 TI - Functional repertoire of dendritic cells generated in granulocyte macrophage colony stimulating factor and interferon-alpha. AB - Monocyte-derived dendritic cells (DCs) generated in granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin-4 (IL-4-DCs) are used to enhance antitumor immunity in cancer patients, although recent evidence suggests that their functional repertoire may be incomplete; in particular, IL-4-DCs appear unable to induce type 2 cytokine-producing T helper (Th) cells. To assess whether type 1 interferon (IFN) could replace IL-4 and generate DCs with a more complete repertoire, we characterized in detail DCs generated from human monocytes cultured with GM-CSF and IFN-alpha (IFN-DCs). We found that IFN-alpha induces DC differentiation more efficiently than IL-4, yielding similar numbers of DCs in a shorter time and that this differentiation persists upon removal of cytokines. Although IFN-DCs had a more mature immunophenotype than IL-4-DCs, showing higher expression of CD80, CD86, and CD83, they still preserved comparable endocytic and phagocytic capacities and responsiveness to maturation stimuli. IFN-DCs had strong antigen-presenting capacity, inducing intense proliferation of T cells to alloantigens or influenza virus. Moreover, IFN-DCs produced lower levels of IL 12p70 and higher levels of IFN-alpha, IL-4, and IL-10 than IL-4-DCs. As a consequence of this different pattern of cytokine secretion, IFN-DCs induced T cells to produce type 1 (IFN-gamma) and type 2 (IL-4 and IL-10) cytokines, and as expected, IL-4-DCs induced only Th1 differentiation. As immune responses with extreme Th1 bias are considered inadequate for the induction of optimal, systemic antitumor immunity, the ability of IFN-DCs to promote more balanced cytokine responses may suggest the advisability to consider these cells in the development of future, DC-based immunotherapy trials. PMID- 14525964 TI - The Src kinase Lyn is a negative regulator of mast cell proliferation. AB - Previous investigators have reported that deletion of the protein tyrosine kinase Lyn alters mast cell (MC) signaling responses but does not affect or reduces the cytokine-mediated proliferation of mouse bone marrow-derived MC (BMMC) precursors and of mature MC. We observed that Lyn-deficient mice have more peritoneal MC than wild-type (WT) mice. Studies to explore this unexpected result showed that Lyn(-/-) BM cells expand faster than WT cells in response to interleukin (IL)-3 and stem-cell factor over the 4-5 weeks required to produce a >95% pure population of granular, receptor with high affinity for immunoglobulin E-positive BMMC. Furthermore, differentiated Lyn(-/-) BMMC continue to proliferate more rapidly than WT BMMC and undergo less apoptosis in response to cytokine withdrawal. Additionally, Lyn(-/-) BMMC support greater IL-3-mediated phosphorylation of the prosurvival kinase, Akt, and the proliferative kinase, extracellular-regulated kinase 1/2. These results identify Lyn as a negative regulator of murine MC survival and proliferation. PMID- 14525965 TI - The interaction of human dendritic cells with yeast and germ-tube forms of Candida albicans leads to efficient fungal processing, dendritic cell maturation, and acquisition of a Th1 response-promoting function. AB - T helper cell type 1 (Th1) cell-mediated immunity plays a critical role in protection against the opportunistic pathogen Candida albicans. Virulence of the fungus is closely associated with its ability to form germ-tubes (GT), the early phase of the dimorphic transition from the commensal yeast (Y) to the more invasive hyphal (H) form. In this study, we examined the functional outcome of the interaction of Y or GT forms with human dendritic cells (DCs), professional antigen-presenting cells, which are pivotal for initiation and modulation of T cell responses. DCs phagocytosed and killed Y and GT cells with a comparable efficiency, becoming able to trigger strong proliferative responses by Candida specific, autologous T cell clones. Both fungal forms induced DC maturation, as indicated by up-regulation of CD83, CD80, CD86, CD40, and major histocompatibility complex classes I and II surface antigens. Chemokine receptors were also modulated in Candida-DCs, which showed increased CCR7/CXCR4 and decreased CCR5 expression. Y- and GT-activated DCs differed in the pattern of cytokine expression. In particular, GT cells, in common with fully differentiated H cells, induced significantly more elevated levels of interleukin (IL)-10 than Y cells. Nevertheless, Y-, GT-, or H-pulsed DCs secreted comparable amounts of IL 12p70. In addition, irrespective of the fungal form triggering DC activation, Candida-DCs acquired the ability to prime naive T lymphocytes with a defined Th1 phenotype. Overall, our findings highlight the induction of substantially similar functional patterns in human DCs encountering the different forms of growth of C. albicans, both seemingly activating the Th1-type immunity which is characteristic of the healthy human subjects, naturally immunized and protected against the fungus. PMID- 14525966 TI - Expression and regulation of antimicrobial peptides in the gastrointestinal tract. AB - The gastrointestinal (GI) tract is exposed to a wide range of microorganisms. The expression of antimicrobial peptides has been demonstrated in different regions of the GI tract, predominantly in epithelial cells, which represent the first host cells with which the microorganisms have to interact for invasion. The intestinal epithelial monolayer is complex, consisting of different cell types, and most have a limited lifespan. Of the GI antimicrobial peptides, alpha- and beta-defensins have been studied the most and are expressed by distinct types of epithelial cells. Enteric alpha-defensin expression is normally restricted to Paneth and intermediate cells in the small intestine. However, there are important differences between mice and humans in the processing of the precursor forms of enteric alpha-defensins. Parasite infection induces an increase in the number of enteric alpha-defensin-expressing Paneth and intermediate cells in the murine small intestine. In the chronically inflamed colonic mucosa, metaplastic Paneth cells (which are absent in the normal colon) also express enteric alpha defensins. Epithelial expression of beta-defensins may be constitutive or inducible by infectious and inflammatory stimuli. The production of some members of the beta-defensin family appears to be restricted to distinct parts of the GI tract. Recent studies using genetically manipulated rodents have demonstrated the likely in vivo importance of enteric antimicrobial peptides in innate host defense against microorganisms. The ability of these peptides to act as chemoattractants for cells of the innate- and adaptive-immune system may also play an important role in perpetuating chronic inflammation in the GI tract. PMID- 14525967 TI - Interferon-gamma: an overview of signals, mechanisms and functions. AB - Interferon-gamma (IFN-gamma) coordinates a diverse array of cellular programs through transcriptional regulation of immunologically relevant genes. This article reviews the current understanding of IFN-gamma ligand, receptor, signal transduction, and cellular effects with a focus on macrophage responses and to a lesser extent, responses from other cell types that influence macrophage function during infection. The current model for IFN-gamma signal transduction is discussed, as well as signal regulation and factors conferring signal specificity. Cellular effects of IFN-gamma are described, including up-regulation of pathogen recognition, antigen processing and presentation, the antiviral state, inhibition of cellular proliferation and effects on apoptosis, activation of microbicidal effector functions, immunomodulation, and leukocyte trafficking. In addition, integration of signaling and response with other cytokines and pathogen-associated molecular patterns, such as tumor necrosis factor-alpha, interleukin-4, type I IFNs, and lipopolysaccharide are discussed. PMID- 14525968 TI - Activation of A2A adenosine receptors inhibits expression of alpha 4/beta 1 integrin (very late antigen-4) on stimulated human neutrophils. AB - The alpha 4/beta 1 integrin very late antigen-4 (CD49d/CD29) is up-regulated on circulating neutrophils of septic patients. Although no individual agent mimics this effect of sepsis, we now report that following priming of human neutrophils with lipopolysaccharide or tumor necrosis factor alpha (TNF-alpha), addition of formyl-Met-Leu-Phe (fMLP) results in a "stimulated", sepsis-like, four- to fivefold rise in CD49d expression. TNF/fMLP stimulation also produced a similar increase in CD49d-mediated adhesion of neutrophils to a vascular cell adhesion molecule-1 (VCAM-1)-coated surface. Adenosine is a naturally occurring, anti inflammatory mediator released from injured or inflamed tissues. We observed that stimulated neutrophil CD49d expression was decreased by activation of A(2A) adenosine receptors (A(2A)AR) with the selective agonist 4-[3-[6-amino-9-(5 ethylcarbamoyl-3,4-dihydroxy-tetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl] cyclohexanecarboxylicacid methyl ester (ATL146e; EC(50)=6.4 nM). ATL146e (100 nM) also reduced the fraction of stimulated neutrophils that adhered to VCAM-1 from 38 +/- 6% to 27 +/- 5%. Inhibition of CD49d expression was equally inhibited by ATL146e, added before or after TNF priming, and was reversed by incubation with the A(2A)AR-selective antagonist 4-[2-[7-amino-2-(2-furyl) (1, 2, 4)triazolo(2,3 a) (1, 3, 5)triazin-5-yl-amino]ethyl]-phenol (ZM241385; 100 nM). A suboptimal ATL146e concentration (1 nM) combined with the type IV phosphodiesterase inhibitor rolipram (100 nM) synergistically decreased stimulated CD49d expression by >50%. The cyclic adenosine monophosphate (cAMP)-dependent kinase [protein kinase A (PKA)] inhibitor H-89 (10 microM) reversed the effect of ATL146e on stimulated CD49d expression. Other means of increasing cAMP in neutrophils also decreased stimulated CD49d expression. We conclude that adenosine binding to A(2A)AR counteracts stimulation of neutrophil CD49d integrin expression and neutrophil binding to VCAM-1 via a cAMP/PKA-mediated pathway. PMID- 14525969 TI - Subset-specific, uniform activation among V gamma 6/V delta 1+ gamma delta T cells elicited by inflammation. AB - The V gamma 6/V delta 1(+) cells, the second murine gamma delta T cell subset to arise in the thymus, express a nearly invariant T cell receptor (TCR), colonize select tissues, and expand preferentially in other tissues during inflammation. These cells are thought to help in regulating the inflammatory response. Until now, V gamma 6/V delta 1(+) cells have only been detectable indirectly, by expression of V gamma 6-encoding mRNA. Here, we report that 17D1, a monoclonal antibody, which detects the related epidermis-associated V gamma 5/V delta 1(+) TCR, will also bind the V gamma 6/V delta 1(+) cells if their TCR is first complexed to an anti-C delta antibody. Features of this special condition for recognition suggest the possibility that an alternate structure exists for the V gamma 6/V delta 1 TCR, which is stabilized upon binding to the anti-C delta antibody. Using the 17D1 antibody as means to track this gamma delta T cell subset by flow cytometry, we discovered that the response of V gamma 6/V delta 1(+) cells during inflammation often far exceeds that of other subsets and that the responding V gamma 6/V delta 1(+) cells display a strikingly uniform activation/memory phenotype compared with other gamma delta T cell subsets. PMID- 14525970 TI - In vitro model for hematopoietic progenitor cell homing reveals endothelial heparan sulfate proteoglycans as direct adhesive ligands. AB - Proteoglycans (PGs) play a dominant role within the bone marrow (BM), but their role in homing of transplanted hematopoietic progenitor cells (HPC) is unknown. In this study, the role of heparan sulfate (HS) PGs on BM endothelium as adhesive structures was investigated. HPC (primary CD34+ cells and cell line KG-1a) were able to bind fractionated heparin, which could be competed by highly sulfated heparin/HS-glycosaminoglycans (GAGs). Under flow conditions, HPC adhered to immobilized heparin after rolling over E-selectin. Rolling of KG-1a on BM endothelial cell (EC) line 4LHBMEC was completely E selectin-dependent. Addition of heparin/HS-GAGs, endothelial treatment with chlorate, or anti-HS all partially inhibited firm adhesion. Moreover, enzymatic removal of endothelial HS-GAGs reduced initial adhesion. Finally, HPC-bound PGs isolated from 4LHBMEC, which was largely inhibited by enzymatic HS-degradation. In summary, we identified sulfated structures on BM endothelium, most likely HSPGs, as a novel class of glycoconjugates involved in the multistep homing cascade of HPC. PMID- 14525971 TI - Analysis of interleukin-1 beta-induced cell signaling activation in rat hippocampus following exposure to gamma irradiation. Protective effect of eicosapentaenoic acid. AB - Among the many reported effects of irradiation in cells is activation of the stress-activated protein kinase, c-Jun N-terminal kinase (JNK), which has been shown to result in apoptotic cell death. The trigger that leads to JNK activation has not been identified, although, in rat hippocampus at least, irradiation induced apoptosis has been coupled with increased accumulation of reactive oxygen species (ROS). Significantly, irradiation-induced changes in hippocampus are abrogated by treatment of rats with the polyunsaturated fatty acid, eicosapentaenoic acid (EPA). A close coupling between ROS accumulation and concentration of the pro-inflammatory cytokine, interleukin-1 beta (IL-1 beta) in hippocampus has been reported, and the evidence suggests that IL-1 beta may be responsible for the enhanced ROS production. Here we set out to assess the possibility that whole body gamma-irradiation increases IL-1 beta concentration in hippocampus and to investigate the consequences of such a change. We present evidence that reveals that the irradiation-induced increase in IL-1 beta concentration in hippocampus is accompanied by increased expression of IL-1 type I receptor and IL-1 accessory protein and increased activation of IL-1 receptor activated kinase. These changes, which were coupled with increased activation of JNK and evidence of apoptotic cell death, were absent in hippocampus of rats that received EPA treatment. Significantly, EPA treatment enhanced hippocampal IL-10 concentration that was inversely correlated with IL-1 beta concentration. The data are consistent with the idea that EPA exerts anti-inflammatory and neuroprotective effects in the central nervous system. PMID- 14525972 TI - The selectivity filter may act as the agonist-activated gate in the G protein activated Kir3.1/Kir3.4 K+ channel. AB - The Kir3.1/Kir3.4 channel is activated by Gbetagamma subunits released on binding of acetylcholine to the M2 muscarinic receptor. A mechanism of channel opening, similar to that for the KcsA and Shaker K+ channels, has been suggested that involves translocation of pore lining transmembrane helices and the opening of an intracellular gate at the "bundle crossing" region. However, in the present study, we show that an extracellular gate at the selectivity filter is critical for agonist activation of the Kir3.1/Kir3.4 channel. Increasing the flexibility of the selectivity filter, by disrupting a salt bridge that lies directly behind the filter, abolished both selectivity for K+ and agonist activation of the channel. Other mutations within the filter that altered selectivity also altered agonist activation. In contrast, mutations within the filter that did not affect selectivity had little if any effect on agonist activation. Interestingly, mutation of bulky side chain phenylalanine residues at the bundle crossing also altered both agonist activation and selectivity. These results demonstrate a significant correlation between agonist activation and selectivity, which is determined by the selectivity filter, and suggests, therefore, that the selectivity filter may act as the agonist-activated gate in the Kir3.1/Kir3.4 channel. PMID- 14525973 TI - Neisserial lipooligosaccharide is a target for complement component C4b. Inner core phosphoethanolamine residues define C4b linkage specificity. AB - We identified Neisseria meningitidis lipooligosaccharide (LOS) as an acceptor for complement component C4b (C4b). Phosphoethanolamine (PEA) residues on the second heptose (HepII) residue in the LOS core structure formed amide linkages with C4b. PEA at the 6-position of HepII (6-PEA) was more efficient than 3-PEA in binding C4b. Strains bearing 6-PEA bound more C4b than strains with 3-PEA and were more susceptible to complement-mediated killing in serum bactericidal assays. Deleting 3-PEA from a strain that expressed both 3- and 6-PEA simultaneously on HepII did not decrease C4b binding. Glycose chain extension of the first heptose residue (HepI) influenced the nature of the C4b-LOS linkage. Predominantly ester C4b-LOS bonds were seen when lacto-N-neotetraose formed the terminus of the glycose chain extension of HepI with 3-PEA on HepII in the LOS core. Related LOS species with more truncated chain extensions from HepI bound C4b via amide linkages to 3-PEA on HepII. However, 6-PEA in the LOS core bound C4b even when the glycose chain from HepI bore lacto-N-neotetraose at the terminus. The C4A isoform exclusively formed amide linkages, whereas C4B bound meningococci preferentially via ester linkages. These data may serve to explain the preponderance of 3-PEA-bearing meningococci among clinical isolates, because 6-PEA enhances C4b binding that may facilitate clearance of 6-PEA-bearing strains resulting from enhanced serum killing by the classical pathway of complement. PMID- 14525974 TI - Resistance to the short term antiproliferative activity of the G-quadruplex ligand 12459 is associated with telomerase overexpression and telomere capping alteration. AB - Ligands that stabilize the telomeric G-rich single-stranded DNA overhang into G quadruplex can be considered as potential antitumor agents that block telomere replication. Ligand 12459, a potent G-quadruplex ligand that belongs to the triazine series, has been previously shown to induce both telomere shortening and apoptosis in the human A549 cell line as a function of its concentration and time exposure. We show here that A549 clones obtained after mutagenesis and selected for resistance to the short term effect of ligand 12459 frequently displayed hTERT transcript overexpression (2-6-fold). Overexpression of hTERT was also characterized in two resistant clones (JFD10 and JFD18) as an increase in telomerase activity, leading to an increase in telomere length. An increased frequency of anaphase bridges was also detected in JFD10 and JFD18, suggesting an alteration of telomere capping functions. Transfection of either hTERT or DN hTERT cDNAs into A549 cells did not confer resistance or hypersensitivity to the short term effect of ligand 12459, indicating that telomerase expression is not the main determinant of the antiproliferative effect of ligand 12459. In contrast, transfection of DN-hTERT cDNA into resistant JFD18 cells restored sensitivity to apoptotic concentrations of ligand 12459, suggesting that telomerase does participate in the resistance to this G-quadruplex ligand. This work provides evidence that telomerase activity is not the main target for the 12459 G-quadruplex ligand but that hTERT functions contribute to the resistance phenotype to this class of agents. PMID- 14525975 TI - Regulation of alpha7 integrin expression during muscle differentiation. AB - Expression of the laminin-binding alpha7 integrin is tightly regulated during myogenic differentiation, reflecting required functions that range from cell motility to formation of stable myotendinous junctions. However, the exact mechanism controlling alpha7 expression in a tissue- and differentiation-specific manner is poorly understood. This report provides evidence that alpha7 gene expression during muscle differentiation is regulated by the c-Myc transcription factor. In myoblasts, alpha7 is expressed at basal levels, but following conversion to myotubes the expression of the integrin is strongly elevated. The increased alpha7 mRNA and protein levels following myogenic differentiation are inversely correlated with c-Myc expression. Transfection of myoblasts with the c Myc transcription factor down-regulated alpha7 expression, whereas overexpression of Madmyc, a dominant-negative c-Myc chimera, induced elevated alpha7 expression. Functional analysis with site-specific deletions identified a specific double E box sequence in the upstream promoter region (-2.0 to -2.6 kb) that is responsible for c-Myc-induced suppression of alpha7 expression. DNA-protein binding assays and supershift analysis revealed that c-Myc forms a complex with this double E-box sequence. Our results suggest that the interaction of c-Myc with this promoter region is an important regulatory element controlling alpha7 integrin expression during muscle development and myotendinous junction formation. PMID- 14525976 TI - Ral GTPases regulate exocyst assembly through dual subunit interactions. AB - Ral GTPases have been implicated in the regulation of a variety of dynamic cellular processes including proliferation, oncogenic transformation, actin cytoskeletal dynamics, endocytosis, and exocytosis. Recently the Sec6/8 complex, or exocyst, a multisubunit complex facilitating post-Golgi targeting of distinct subclasses of secretory vesicles, has been identified as a bona fide Ral effector complex. Ral GTPases regulate exocyst-dependent vesicle trafficking and are required for exocyst complex assembly. Sec5, a membrane-associated exocyst subunit, has been identified as a direct target of activated Ral; however, the mechanism by which Ral can modulate exocyst assembly is unknown. Here we report that an additional component of the exocyst, Exo84, is a direct target of activated Ral. We provide evidence that mammalian exocyst components are present as distinct subcomplexes on vesicles and the plasma membrane and that Ral GTPases regulate the assembly interface of a full octameric exocyst complex through interaction with Sec5 and Exo84. PMID- 14525978 TI - Human MUC2 mucin gene is transcriptionally regulated by Cdx homeodomain proteins in gastrointestinal carcinoma cell lines. AB - In intestinal metaplasia and 30% of gastric carcinomas, MUC2 intestinal mucin and the intestine-specific transcription factors Cdx-1 and Cdx-2 are aberrantly expressed. The involvement of Cdx-1 and Cdx-2 in the intestinal development and their role in transcription of several intestinal genes support the hypothesis that Cdx-1 and/or Cdx-2 play important roles in the aberrant intestinal differentiation program of intestinal metaplasia and gastric carcinoma. To clarify the mechanisms of transcriptional regulation of the MUC2 mucin gene in gastric cells, pGL3 deletion constructs covering 2.6 kb of the human MUC2 promoter were used in transient transfection assays, enabling us to identify a relevant region for MUC2 transcription in all gastric cell lines. To evaluate the role of Cdx-1 and Cdx-2 in MUC2 transcription we performed co-transfection experiments with expression vectors encoding Cdx-1 and Cdx-2. In two of the four gastric carcinoma cell lines and in all colon carcinoma cell lines we observed transactivation of the MUC2 promoter by Cdx-2. Using gel shift assays we identified two Cdx-2 binding sites at -177/-171 and -191/-187. Only simultaneous mutation of the two sites resulted in inhibition of Cdx-2-mediated transactivation of MUC2 promoter, implying that both Cdx-2 sites are active. Finally, stable expression of Cdx-2 in a gastric cell line initially not expressing Cdx-2, led to induction of MUC2 expression. In conclusion, this work demonstrates that Cdx-2 activates the expression of MUC2 mucin gene in gastric cells, inducing an intestinal transdifferentiation phenotype that parallels what is observed both in intestinal metaplasia and some gastric carcinomas. PMID- 14525977 TI - Combining Fas mutation with interleukin-2 deficiency prevents Colitis and Lupus: implicating interleukin-2 for auto-reactive T cell expansion and Fas ligand for colon epithelial cell death. AB - Both the lpr gene defect and interleukin 2-targeted mutation (IL-2 KO) in mice are lethal. Interestingly, mice bearing both mutations live significantly longer than mice with either of the single mutant genes, approximating the life span of normal controls. They do not display the major disease phenotypes of lpr and IL-2 KO mice. Systemic autoimmune response, the accumulation of the abnormal CD4-CD8 B220+ double-negative T cells, kidney disease pathology, anemia, colon damage, and lethality are prevented. Our data indicate that IL-2 is mandatory for the expansion of auto-reactive T cells in lpr mice and that CD95 (Fas) is the critical target for the development of anemia and ulcerative colitis in IL-2 KO mice in which CD178 (FasL) on intraepithelial T cells is the major effector responsible for colon damage and lethality. PMID- 14525979 TI - Structure-function analysis of Trypanosoma brucei RNA triphosphatase and evidence for a two-metal mechanism. AB - Trypanosoma brucei RNA triphosphatase TbCet1 is a 252-amino acid polypeptide that catalyzes the first step in mRNA cap formation. By performing an alanine scan of TbCet1, we identified six amino acids that are essential for triphosphatase activity (Glu-52, Arg-127, Glu-168, Arg-186, Glu-216, and Glu-218). These results consolidate the proposal that protozoan, fungal, and Chlorella virus RNA triphosphatases belong to a single family of metal-dependent NTP phosphohydrolases with a unique tunnel active site composed of eight beta strands. Limited proteolysis of TbCet1 suggests that the hydrophilic N terminus is surface-exposed, whereas the catalytic core domain is tightly folded with the exception of a protease-sensitive loop (76WKGRRARKT84) between two of the putative tunnel strands. The catalytic domain of TbCet1 is extraordinarily thermostable. It remains active after heating for 2 h at 75 degrees C. Analysis by zonal velocity sedimentation indicates that TbCet1 is a monomeric enzyme, unlike fungal RNA triphosphatases, which are homodimers. We show that tripolyphosphate is a potent competitive inhibitor of TbCet1 (Ki 1.4 microm) that binds more avidly to the active site than the ATP substrate (Km 25 microm). We present evidence of synergistic activation of the TbCet1 triphosphatase by manganese and magnesium, consistent with a two-metal mechanism of catalysis. Our findings provide new insight to the similarities (in active site tertiary structure and catalytic mechanism) and differences (in quaternary structure and thermal stability) among the different branches of the tunnel enzyme family. PMID- 14525980 TI - Expression of a human surfactant protein C mutation associated with interstitial lung disease disrupts lung development in transgenic mice. AB - Surfactant Protein C (SP-C) is a secreted transmembrane protein that is exclusively expressed by alveolar type II epithelial cells of the lung. SP-C associates with surfactant lipids to reduce surface tension within the alveolus, maintaining lung volume at end expiration. Mutations in the gene encoding SP-C (SFTPC) have recently been linked to chronic lung disease in children and adults. The goal of this study was to determine whether a disease-linked mutation in SFTPC causes lung disease in transgenic mice. The SFTPC mutation, designated g.1728 G --> A, results in the deletion of exon4, generating a truncated form of SP-C (SP-C(Deltaexon4)). cDNA encoding SP-C(Deltaexon4) was constitutively expressed in type II epithelial cells of transgenic mice. Viable F0 transgene positive mice were not generated after two separate rounds of pronuclear injections. Histological analysis of lung tissue harvested from embryonic day 17.5 F0 transgene-positive fetuses revealed that SP-C(Deltaexon4) caused a dose dependent disruption in branching morphogenesis of the lung associated with epithelial cell cytotoxicity. Transient expression of SP-C(Deltaexon4) in isolated type II epithelial cells or HEK293 cells resulted in incomplete processing of the mutant proprotein, a dose-dependent increase in BiP transcription, trapping of the proprotein in the endoplasmic reticulum, and rapid degradation via a proteasome-dependent pathway. Taken together, these data suggest that the g.1728 G --> A mutation causes misfolding of the SP-C proprotein with subsequent induction of the unfolded protein response and endoplasmic reticulum-associated degradation pathways ultimately resulting in disrupted lung morphogenesis. PMID- 14525981 TI - Tup1-Ssn6 interacts with multiple class I histone deacetylases in vivo. AB - The Tup1-Ssn6 corepressor complex in Saccharomyces cerevisiae represses the transcription of a diverse set of genes. Chromatin is an important component of Tup1-Ssn6-mediated repression. Tup1 binds to underacetylated histone tails and requires multiple histone deacetylases (HDACs) for its repressive functions. Here, we describe physical interactions of the corepressor complex with the class I HDACs Rpd3, Hos2, and Hos1. In contrast, no in vivo interaction was observed between Tup-Ssn6 and Hda1, a class II HDAC. We demonstrate that Rpd3 interacts with both Tup1 and Ssn6. Rpd3 and Hos2 interact with Ssn6 independently of Tup1 via distinct tetratricopeptide domains within Ssn6, suggesting that these two HDACs may contact the corepressor at the same time. PMID- 14525983 TI - DACH1 inhibits transforming growth factor-beta signaling through binding Smad4. AB - The vertebrate homologues of Drosophila dachsund, DACH1 and DACH2, have been implicated as important regulatory genes in development. DACH1 plays a role in retinal and pituitary precursor cell proliferation and DACH2 plays a specific role in myogenesis. DACH proteins contain a domain (DS domain) that is conserved with the proto-oncogenes Ski and Sno. Since the Ski/Sno proto-oncogenes repress AP-1 and SMAD signaling, we hypothesized that DACH1 might play a similar cellular function. Herein, DACH1 was found to be expressed in breast cancer cell lines and to inhibit transforming growth factor-beta (TGF-beta)-induced apoptosis. DACH1 repressed TGF-beta induction of AP-1 and Smad signaling in gene reporter assays and repressed endogenous TGF-beta-responsive genes by microarray analyses. DACH1 bound to endogenous NCoR and Smad4 in cultured cells and DACH1 co-localized with NCoR in nuclear dotlike structures. NCoR enhanced DACH1 repression, and the repression of TGF-beta-induced AP-1 or Smad signaling by DACH1 required the DACH1 DS domain. The DS domain of DACH was sufficient for NCoR binding at a Smad4 binding site. Smad4 was required for DACH1 repression of Smad signaling. In Smad4 null HTB-134 cells, DACH1 inhibited the activation of SBE-4 reporter activity induced by Smad2 or Smad3 only in the presence of Smad4. DACH1 participates in the negative regulation of TGF-beta signaling by interacting with NCoR and Smad4. PMID- 14525982 TI - Reversible topological organization within a polytopic membrane protein is governed by a change in membrane phospholipid composition. AB - Once inserted, transmembrane segments of polytopic membrane proteins are generally considered stably oriented due to the large free energy barrier to topological reorientation of adjacent extramembrane domains. However, the topology and function of the polytopic membrane protein lactose permease of Escherichia coli are dependent on the membrane phospholipid composition, revealing topological dynamics of transmembrane domains after stable membrane insertion (Bogdanov, M., Heacock, P. N., and Dowhan, W. (2002) EMBO J. 21, 2107 2116). In this study, we show that the high affinity phenylalanine permease PheP shares many similarities with lactose permease. PheP assembled in a mutant of E. coli lacking phosphatidylethanolamine (PE) exhibited significantly reduced active transport function and a complete inversion in topological orientation of the N terminus and adjoining transmembrane hairpin loop compared with PheP in a PE containing strain. Introduction of PE following the assembly of PheP triggered a reorientation of the N terminus and adjacent hairpin to their native orientation associated with regain of wild-type transport function. The reversible orientation of these secondary transport proteins in response to a change in phospholipid composition might be a result of inherent conformational flexibility necessary for transport function or during protein assembly. PMID- 14525984 TI - Identification and characterization of an amphipathic leucine zipper-like motif in Escherichia coli toxin hemolysin E. Plausible role in the assembly and membrane destabilization. AB - Hemolysin E (HlyE) is a 34 kDa protein toxin, recently isolated from a pathogenic strain of Escherichia coli, which is believed to exert its toxic activity via formation of pores in the target cell membrane. With the goal of understanding the involvement of different segments of hemolysin E in the membrane interaction and assembly of the toxin, a conserved, amphipathic leucine zipper-like motif has been identified. In order to evaluate the possible structural and functional roles of this segment in HlyE, a 30-residue peptide (H-205) corresponding to the leucine zipper motif (amino acid 205-234) and two mutant peptides of the same size were synthesized and labeled by fluorescent probes at their N termini. The results show that the wild-type H-205 binds to both zwitterionic (PC/Chol) and negatively charged (PC/PG/Chol) phospholipid vesicles and also self-assemble therein. Detailed membrane-binding experiments revealed that this synthetic motif (H-205) formed large aggregates and inserted into the bilayer of only negatively charged lipid vesicles but not of zwitterionic membrane. Although both the mutants bound to zwitterionic and negatively charged lipid vesicles, neither of them inserted into the lipid bilayers nor assembled in any of these lipid vesicles. Furthermore, H-205 adopted a significant helical structure in membrane mimetic environments and induced the permeation of monovalent ions and release of entrapped calcein across the phospholipid vesicles more efficiently than the mutant peptides. The results presented here indicate that this H-205 (amino acid 205-234) segment may be an important structural element in hemolysin E, which could play a significant role in the binding and assembly of the toxin in the target cell membrane and its destabilization. PMID- 14525985 TI - Solution structure of the dimeric zinc binding domain of the chaperone ClpX. AB - ClpX (423 amino acids), a member of the Clp/Hsp100 family of molecular chaperones and the protease, ClpP, comprise a multimeric complex supporting targeted protein degradation in Escherichia coli. The ClpX sequence consists of an NH2-terminal zinc binding domain (ZBD) and a COOH-terminal ATPase domain. Earlier, we have demonstrated that the zinc binding domain forms a constitutive dimer that is essential for the degradation of some ClpX substrates such as gammaO and MuA but is not required for the degradation of other substrates such as green fluorescent protein-SsrA. In this report, we present the NMR solution structure of the zinc binding domain dimer. The monomer fold reveals that ZBD is a member of the treble clef zinc finger family, a motif known to facilitate protein-ligand, protein-DNA, and protein-protein interactions. However, the dimeric ZBD structure is not related to any protein structure in the Protein Data Bank. A trimer-of-dimers model of ZBD is presented, which might reflect the closed state of the ClpX hexamer. PMID- 14525986 TI - Cut5 is required for the binding of Atr and DNA polymerase alpha to genotoxin damaged chromatin. AB - DNA damage triggers the assembly of checkpoint signaling proteins on chromatin that activate the Chk1 signaling pathway and block S-phase progression. Here we show that genotoxin-induced Chk1 activation requires Cut5 (Mus101/TopBP1) in a process that is independent of the role of Cut5 in DNA replication. Analysis of the role of Cut5 in checkpoint activation revealed that it associated with chromatin following DNA damage in a process that required RPA. Additionally, Cut5 was required for the recruitment of Atr, DNA polymerase alpha, and Rad1 but not RPA to chromatin following DNA damage. Taken together, these results demonstrate that Cut5 plays an integral role in the recruitment and assembly of the Chk1 signaling cascade components following DNA damage. PMID- 14525987 TI - Structure, function, and regulation of a subfamily of mouse zinc transporter genes. AB - Zinc is an essential metal for all eukaryotes, and cells have evolved a complex system of proteins to maintain the precise balance of zinc uptake, intracellular storage, and efflux. In mammals, zinc uptake appears to be mediated by members of the Zrt/Irt-like protein (ZIP) superfamily of metal ion transporters. Herein, we have studied a subfamily of zip genes (zip1, zip2, and zip3) that is conserved in mice and humans. These eight-transmembrane domain proteins contain a conserved 12 amino acid signature sequence within the fourth transmembrane domain. All three of these mouse ZIP proteins function to specifically increase the uptake of zinc in transfected cultured cells, similar to the previously demonstrated functions of human ZIP1 and ZIP2 (Gaither, L. A., and Eide, D. J. (2000) J. Biol. Chem. 275, 5560-5564; Gaither, L. A., and Eide, D. J. (2001) J. Biol. Chem. 276, 22258 22264). No ZIP3 orthologs have been previously studied. Furthermore, this first systematic comparative study of the in vivo expression and dietary zinc regulation of this subfamily of zip genes revealed that 1) zip1 mRNA is abundant in many mouse tissues, whereas zip2 and zip3 mRNAs are very rare or moderately rare, respectively, and tissue-restricted in their accumulation; and 2) unlike mouse metallothionein I and zip4 mRNAs (Dufner-Beattie, J., Wang, F., Kuo, Y.-M., Gitschier, J., Eide, D., and Andrews, G. K. (2003) J. Biol. Chem. 278, 33474 33481), the abundance of zip1, zip2, and zip3 mRNAs is not regulated by dietary zinc in the intestine and visceral endoderm, tissues involved in nutrient absorption. These studies suggest that all three of these ZIP proteins may play cell-specific roles in zinc homeostasis rather than primary roles in the acquisition of dietary zinc. PMID- 14525988 TI - Closely related G-protein-coupled receptors use multiple and distinct domains on G-protein alpha-subunits for selective coupling. AB - The molecular basis of selectivity in G-protein receptor coupling has been explored by comparing the abilities of G-protein heterotrimers containing chimeric Galpha subunits, comprised of various regions of Gi1alpha, Gtalpha, and Gqalpha, to stabilize the high affinity agonist binding state of serotonin, adenosine, and muscarinic receptors. The data indicate that multiple and distinct determinants of selectivity exist for individual receptors. While the A1 adenosine receptor does not distinguish between Gi1alpha and Gtalpha sequences, the 5-HT1A and 5-HT1B serotonin and M2 muscarinic receptors can couple with Gi1 but not Gt. It is possible to distinguish domains that eliminate coupling and are defined as "critical," from those that impair coupling and are defined as "important." Domains within the N terminus, alpha4-helix, and alpha4-helix alpha4/beta6-loop of Gi1alpha are involved in 5-HT and M2 receptor interactions. Chimeric Gi1alpha/Gqalpha subunits verify the critical role of the Galpha C terminus in receptor coupling, however, the individual receptors differ in the C terminal amino acids required for coupling. Furthermore, the EC50 for interactions with Gi1 differ among the individual receptors. These results suggest that coupling selectivity ultimately involves subtle and cooperative interactions among various domains on both the G-protein and the associated receptor as well as the G-protein concentration. PMID- 14525989 TI - Inhibition of receptor-mediated endocytosis demonstrates generation of amyloid beta-protein at the cell surface. AB - Sequential cleavages of the amyloid beta-protein precursor (APP) by the beta- and gamma-secretases generate the amyloid beta-protein (A beta), which plays a central role in Alzheimer's disease. Previous work provided evidence for involvement of both the secretory and endocytic pathways in A beta generation. Here, we used HeLa cells stably expressing a tetracycline-regulated dominant negative dynamin I (dyn K44A), which selectively inhibits receptor-mediated endocytosis, and analyzed the effects on the processing of endogenous APP. Upon induction of dyn K44A, levels of mature APP rose at the cell surface, consistent with retention of APP on the plasma membrane. The alpha-secretase cleavage products of APP were increased by dyn K44A, in that alpha-APPs in medium and the C83 C-terminal stub in the membrane both rose. The beta-secretase cleavage of APP, C99, also increased modestly. The use of specific gamma-secretase inhibitors to study the accumulation of alpha- and beta-cleavage products independent of their processing by gamma-secretase confirmed that retention of APP on the plasma membrane results in increased processing by both alpha- and beta-secretases. Unexpectedly, endogenous A beta secretion was significantly increased by dyn K44A, as detected by three distinct methods: metabolic labeling, immunoprecipitation/Western blotting, and enzyme-linked immunosorbent assay. Levels of p3 (generated by sequential alpha- and gamma-cleavage) also rose. We conclude that endogenous A beta can be produced directly at the plasma membrane and that alterations in the degree of APP endocytosis may help regulate its production. Our findings are consistent with a role for the gamma-secretase complex in the processing of numerous single-transmembrane receptors at the cell surface. PMID- 14525990 TI - Role of charged residues in coupling ligand binding and channel activation in the extracellular domain of the glycine receptor. AB - The glycine receptor is a member of the ligand-gated ion channel receptor superfamily that mediates fast synaptic transmission in the brainstem and spinal cord. Following ligand binding, the receptor undergoes a conformational change that is conveyed to the transmembrane regions of the receptor resulting in the opening of the channel pore. Using the acetylcholine-binding protein structure as a template, we modeled the extracellular domain of the glycine receptor alpha1 subunit and identified the location of charged residues within loops 2 and 7 (the conserved Cys-loop). These loops have been postulated to interact with the M2-M3 linker region between the transmembrane domains 2 and 3 as part of the receptor activation mechanism. Charged residues were substituted with cysteine, resulting in a shift in the concentration-response curves to the right in each case. Covalent modification with 2-(trimethylammonium) ethyl methanethiosulfonate was demonstrated only for K143C, which was more accessible in the open state than the closed state, and resulted in a shift in the EC50 toward wild-type values. Charge reversal mutations (E53K, D57K, and D148K) also impaired channel activation, as inferred from increases in EC50 values and the conversion of taurine from an agonist to an antagonist in E53K and D57K. Thus, each of the residues Glu-53, Asp 57, Lys-143, and Asp-148 are implicated in channel gating. However, the double reverse charge mutations E53K:K276E, D57K:K276E, and D148K:K276E did not restore glycine receptor function. These results indicate that loops 2 and 7 in the extracellular domain play an important role in the mechanism of activation of the glycine receptor although not by a direct electrostatic mechanism. PMID- 14525991 TI - Mechanism and molecular determinant for regulation of rabbit transient receptor potential type 5 (TRPV5) channel by extracellular pH. AB - The transient receptor potential type 5 (TRPV5) channel is present in kidney and intestine and important for transepithelial (re)absorption of calcium in these tissues. We report that in whole-cell patch clamp recording extracellular acidification inhibited rabbit TRPV5 with apparent pKa approximately 6.55. The two extracellular loops between the fifth and sixth transmembrane segments of TRPV5 presumably form part of the outer opening of the pore and likely are important in binding and regulation by external protons. We found that mutation of glutamate 522 to glutamine (E522Q) decreased the sensitivity of the channel to extracellular acidification. Mutations of other titratable amino acids within the two extracellular loops to non-titratable amino acids had no effect on pH sensitivity. Substitutions of aspartate or other titratable amino acids for glutamate 522 conferred an increase in pH sensitivity. The pH sensitivity mediated by glutamate 522 was independent of extracellular or intracellular Mg2+. Single channel analysis revealed that extracellular acidification reduced single channel conductance as well as open probability of the wild type channel. In contrast to wild type channel, extracellular acidification did not reduce open probability for E522Q mutant. Methanethiosulfonate reagents inhibited the activity of glutamine 522 to cysteine mutant channel with a reaction rate constant approaching that with free thiols in solution, suggesting that glutamate 522 is located on the surface of the channel. These data suggest that glutamate 522 of the rabbit TRPV5 is a "pH sensor," and extracellular protons inhibit TRPV5 likely by altering conformation of the channel protein. PMID- 14525992 TI - Cloning and characterization of Xen-dorphin prohormone from Xenopus laevis: a new opioid-like prohormone distinct from proenkephalin and prodynorphin. AB - Opioid-like peptides mediate analgesia and induce behavioral effects such as tolerance and dependence by ligand-receptor-mediated mechanisms. The classical opioid prohormones can generate several bioactive peptides, and these divergent families of prohormones share a common well conserved ancestral opioid motif (Tyr Gly-Gly-Phe). Evidence from pharmacological and molecular cloning studies indicates the presence of multiple isoforms of opioid ligands and receptors that are as yet uncharacterized. To identify potential new members we used the opioid motif as an anchor sequence and isolated two distinct isoforms (Xen-dorphins A and B) of an opioid prohormone from Xenopus laevis brain cDNA library. Xen dorphin prohormones can generate multiple novel opioid ligands distinct from the known members of this family. Both isoforms are present in a wide variety of tissues including the brain. Two potential bioactive peptides, Xen-dorphin-1A and -1B, that were chemically synthesized showed opioid agonist activity in frog and rat brain membranes using a [35S]GTPgammaS assay. Initial radioligand binding experiments demonstrated that Xen-dorphin-1B binds with high affinity to opioid receptor(s) and with potential preference to the kappa-opioid receptor subtype. Cloning of the Xen-dorphin prohormone provides new evidence for the potential presence of other members in the opioid peptide superfamily. PMID- 14525993 TI - Modification of activity and specificity of haloalkane dehalogenase from Sphingomonas paucimobilis UT26 by engineering of its entrance tunnel. AB - Structural comparison of three different haloalkane dehalogenases suggested that substrate specificity of these bacterial enzymes could be significantly influenced by the size and shape of their entrance tunnels. The surface residue leucine 177 positioned at the tunnel opening of the haloalkane dehalogenase from Sphingomonas paucimobilis UT26 was selected for modification based on structural and phylogenetic analysis; the residue partially blocks the entrance tunnel, and it is the most variable pocket residue in haloalkane dehalogenase-like proteins with nine substitutions in 14 proteins. Mutant genes coding for proteins carrying all possible substitutions in position 177 were constructed by site-directed mutagenesis and heterologously expressed in Escherichia coli. In total, 15 active protein variants were obtained, suggesting a relatively high tolerance of the site for the introduction of mutations. Purified protein variants were kinetically characterized by determination of specific activities with 12 halogenated substrates and steady-state kinetic parameters with two substrates. The effect of mutation on the enzyme activities varied dramatically with the structure of the substrates, suggesting that extrapolation of one substrate to another may be misleading and that a systematic characterization of the protein variants with a number of substrates is essential. Multivariate analysis of activity data revealed that catalytic activity of mutant enzymes generally increased with the introduction of small and nonpolar amino acid in position 177. This result is consistent with the phylogenetic analysis showing that glycine and alanine are the most commonly occurring amino acids in this position among haloalkane dehalogenases. The study demonstrates the advantages of using rational engineering to develop enzymes with modified catalytic properties and substrate specificities. The strategy of using site-directed mutagenesis to modify a specific entrance tunnel residue identified by structural and phylogenetic analyses, rather than combinatorial screening, generated a high percentage of viable mutants. PMID- 14525994 TI - Spheniscins, avian beta-defensins in preserved stomach contents of the king penguin, Aptenodytes patagonicus. AB - During the last part of egg incubation in king penguins, the male can preserve undigested food in the stomach for several weeks. This ensures survival of the newly hatched chick, in cases where the return of the foraging female from the sea is delayed. In accordance with the characterization of stress-induced bacteria, we demonstrate the occurrence of strong antimicrobial activities in preserved stomach contents. We isolated and fully characterized two isoforms of a novel 38-residue antimicrobial peptide (AMP), spheniscin, belonging to the beta defensin subfamily. Spheniscin concentration was found to strongly increase during the period of food storage. Using a synthetic version of one of two spheniscin isoforms, we established that this peptide has a broad activity spectrum, affecting the growth of both pathogenic bacteria and fungi. Altogether, our data suggest that spheniscins and other, not yet identified, antimicrobial substances may play a role in the long term preservation of stored food in the stomach of king penguins. PMID- 14525995 TI - Thrombin activatable fibrinolysis inhibitor, a potential regulator of vascular inflammation. AB - The latent plasma carboxypeptidase thrombin-activable fibrinolysis inhibitor (TAFI) is activated by thrombin/thrombomodulin on the endothelial cell surface, and functions in dampening fibrinolysis. In this study, we examined the effect of activated TAFI (TAFIa) in modulating the proinflammatory functions of bradykinin, complement C5a, and thrombin-cleaved osteopontin. Hydrolysis of bradykinin and C5a and thrombin-cleaved osteopontin peptides by TAFIa was as efficient as that of plasmin-cleaved fibrin peptides, indicating that these are also good substrates for TAFIa. Plasma carboxypeptidase N, generally regarded as the physiological regulator of kinins, was much less efficient than TAFIa. TAFIa abrogated C5a-induced neutrophil activation in vitro. Jurkat cell adhesion to osteopontin was markedly enhanced by thrombin cleavage of osteopontin. This was abolished by TAFIa treatment due to the removal of the C-terminal Arg168 by TAFIa from the exposed SVVYGLR alpha 4 beta 1 integrin-binding site in thrombin-cleaved osteopontin. Thus, thrombin cleavage of osteopontin followed by TAFIa treatment may sequentially up- and down-modulate the pro-inflammatory properties of osteopontin. An engineered anticoagulant thrombin, E229K, was able to activate endogenous plasma TAFI in mice, and E229K thrombin infusion effectively blocked bradykinin-induced hypotension in wild-type, but not in TAFI-deficient, mice in vivo. Our data suggest that TAFIa may have a broad anti-inflammatory role, and its function is not restricted to fibrinolysis. PMID- 14525996 TI - Folding intermediates of the prion protein stabilized by hydrostatic pressure and low temperature. AB - Prion diseases are associated with conformational conversion of the cellular prion protein, PrPC, into a misfolded form, PrPSc. We have investigated the equilibrium unfolding of the structured domain of recombinant murine prion protein, comprising residues 121-231 (mPrP-(121-231)). The equilibrium unfolding of mPrP-(121-231) by urea monitored by intrinsic fluorescence and circular dichroism (CD) spectroscopies indicated a two-state transition, without detectable folding intermediates. The fluorescent probe 4,4'-dianilino-1,1' binaphthyl-5,5-disulfonic acid (bis-ANS) binds to native mPrP-(121-231), indicating exposure of hydrophobic domains on the protein surface. Increasing concentrations of urea (up to 4 M) caused the release of bound bis-ANS, whereas changes in intrinsic fluorescence and CD of mPrP took place only above 4 M urea. This indicates the existence of a partially unfolded conformation of mPrP, characterized by loss of bis-ANS binding and preservation of the overall structure of the protein, stabilized at low concentrations of urea. Hydrostatic pressure and low temperatures were also used to stabilize partially folded intermediates that are not detectable in the presence of chemical denaturants. Compression of mPrP to 3.5 kbar at 25 degrees C and pH 7 caused a slight decrease in intrinsic fluorescence emission and an 8-fold increase in bis-ANS fluorescence. Lowering the temperature to -9 degrees C under pressure reversed the decrease in intrinsic fluorescence and caused a marked (approximately 40 fold) increase in bis-ANS fluorescence. The increase in bis-ANS fluorescence at low temperatures was similar to that observed for mPrP at 1 atm at pH 4. These results suggest that pressure-assisted cold denaturation of mPrP stabilizes a partially folded intermediate that is qualitatively similar to the state obtained at acidic pH. Compression of mPrP in the presence of a subdenaturing concentration of urea stabilized another partially folded intermediate, and cold denaturation under these conditions led to complete unfolding of the protein. Possible implications of the existence of such partially folded intermediates in the folding of the prion protein and in the conversion to the PrPSc conformer are discussed. PMID- 14525997 TI - Recombinant expression and characterization of a novel fibronectin isoform expressed in cartilaginous tissues. AB - A novel fibronectin (FN) isoform lacking the segment from IIICS (type III connecting segment) through the I-10 module is expressed predominantly in normal cartilaginous tissues. We expressed and purified recombinant cartilage-type FN using a mammalian expression system and characterized its molecular and biological properties. Although FNs have been shown to be secreted as disulfide bonded dimers, cartilage-type FN was secreted mainly as a monomer. It was less potent than plasma-type FN in promoting cell adhesion and binding to integrin alpha5beta1, although it was more active than plasma-type FN in binding to chondroitin sulfate E. When added exogenously, cartilage-type FN was poorly assembled into the fibrillar FN matrix, mostly because of its monomeric structure. Given that cartilage is characterized by its non-fibrillar matrix with abundant chondroitin sulfate-containing proteoglycans, it is likely that cartilage-type FN has evolved to adapt itself to the non-fibrillar structure of the cartilage matrix through acquisition of a novel mechanism of alternative pre mRNA splicing. PMID- 14525998 TI - Glycation and post-translational processing of human interferon-gamma expressed in Escherichia coli. AB - Until recently, nonenzymatic glycosylation (glycation) was thought to affect the proteins of long living eukaryotes only. However, in a recent study (Mironova, R., Niwa, T., Hayashi, H., Dimitrova, R., and Ivanov, I. (2001) Mol. Microbiol. 39, 1061-1068), we have shown that glycation takes place in Escherichia coli as well. In the present study, we demonstrate that the post-translational processing (proteolysis and covalent dimerization) observed with cysteineless recombinant human interferon-gamma (rhIFN-gamma) is tightly associated with its in vivo glycation. Our results show that, at the time of isolation, rhIFN-gamma contained early (but not advanced) glycation products. Using reverse phase high performance liquid chromatography in conjunction with fluorescence measurements, enzyme linked immunosorbent assay, and mass spectrometry, we found that advanced glycation end products arose in rhIFN-gamma during storage. The latter were identified mainly in the Arg/Lys-rich C terminus of the protein, which was also the main target of proteolysis. Mass spectral analysis and N-terminal sequencing revealed four major (Arg140/Arg141, Phe137/Arg138, Met135/Leu136, and Lys131/Arg132) and two minor (Lys109/Ala110 and Arg90/Asp91) cleavage sites in this region. Tryptic peptide mapping indicated that the covalent dimers of rhIFN gamma originating during storage were formed mainly by lateral cross-linking of the monomer subunits. Antiviral assay showed that proteolysis lowered the antiviral activity of rhIFN-gamma, whereas covalent dimerization completely abolished it. PMID- 14525999 TI - DNA lesion recognition by the bacterial repair enzyme MutM. AB - MutM is a bacterial DNA glycosylase that removes the mutagenic lesion 8 oxoguanine (oxoG) from duplex DNA. The means of oxoG recognition by MutM (also known as Fpg) is of fundamental interest, in light of the vast excess of normal guanine bases present in genomic DNA. The crystal structure of a recognition competent but catalytically inactive version of MutM in complex with oxoG containing DNA reveals the structural basis for recognition. MutM binds the oxoG nucleoside in the syn glycosidic configuration and distinguishes oxoG from guanine by reading out the protonation state of the N7 atom. The segment of MutM principally responsible for oxoG recognition is a flexible loop, suggesting that conformational mobility influences lesion recognition and catalysis. Furthermore, the structure of MutM in complex with DNA containing an alternative substrate, dihydrouracil, demonstrates how MutM is able to recognize lesions other than oxoG. PMID- 14526000 TI - Cellulosomes from mesophilic bacteria. PMID- 14526001 TI - Production of high-quality particulate methane monooxygenase in high yields from Methylococcus capsulatus (bath) with a hollow-fiber membrane bioreactor. AB - In order to obtain particulate methane monooxygenase (pMMO)-enriched membranes from Methylococcus capsulatus (Bath) with high activity and in high yields, we devised a method to process cell growth in a fermentor adapted with a hollow fiber bioreactor that allows easy control and quantitative adjustment of the copper ion concentration in NMS medium over the time course of cell culture. This technical improvement in the method for culturing bacterial cells allowed us to study the effects of copper ion concentration in the growth medium on the copper content in the membranes, as well as the specific activity of the enzyme. The optimal copper concentration in the growth medium was found to be 30 to 35 micro M. Under these conditions, the pMMO is highly expressed, accounting for 80% of the total cytoplasmic membrane proteins and having a specific activity as high as 88.9 nmol of propylene oxide/min/mg of protein with NADH as the reductant. The copper stoichiometry is approximately 13 atoms per pMMO molecule. Analysis of other metal contents provided no evidence of zinc, and only traces of iron were present in the pMMO-enriched membranes. Further purification by membrane solubilization in dodecyl beta-D-maltoside followed by fractionation of the protein-detergent complexes according to molecular size by gel filtration chromatography resulted in a good yield of the pMMO-detergent complex and a high level of homogeneity. The pMMO-detergent complex isolated in this way had a molecular mass of 220 kDa and consisted of an alphabetagamma protein monomer encapsulated in a micelle consisting of ca. 240 detergent molecules. The enzyme is a copper protein containing 13.6 mol of copper/mol of pMMO and essentially no iron (ratio of copper to iron, 80:1). Both the detergent-solubilized membranes and the purified pMMO-detergent complex exhibited reasonable, if not excellent, specific activity. Finally, our ability to control the level of expression of the pMMO allowed us to clarify the sensitivity of the enzyme to NADH and duroquinol, the two common reductants used to assay the enzyme. PMID- 14526002 TI - Redefining the role of psr in beta-lactam resistance and cell autolysis of Enterococcus hirae. AB - The contribution of penicillin-binding protein 5 (PBP5) and the PBP5 synthesis repressor (Psr) to the beta-lactam resistance, growth, and cell autolysis of wild type strain ATCC 9790 and resistant strain R40 of Enterococcus hirae was investigated by disruption or substitution of the corresponding pbp5 and psr genes by Campbell-type recombination. The resulting modifications were confirmed by hybridization and PCR. The low susceptibility of E. hirae to beta-lactams was confirmed to be largely dependent on the presence of PBP5. However, against all expectations, inactivation of psr in ATCC 9790 or complementation of R40 cells with psr did not modify the susceptibility to benzylpenicillin or the growth and cell autolysis rates. These results indicated that the psr gene does not seem to be involved in the regulation of PBP5 synthesis and consequently in beta-lactam resistance or in the regulation of cell autolysis in E. hirae. PMID- 14526003 TI - Cloning and characterization of archaeal type I signal peptidase from Methanococcus voltae. AB - Archaeal protein trafficking is a poorly characterized process. While putative type I signal peptidase genes have been identified in sequenced genomes for many archaea, no biochemical data have been presented to confirm that the gene product possesses signal peptidase activity. In this study, the putative type I signal peptidase gene in Methanococcus voltae was cloned and overexpressed in Escherichia coli, the membranes of which were used as the enzyme source in an in vitro peptidase assay. A truncated, His-tagged form of the M. voltae S-layer protein was generated for use as the substrate to monitor the signal peptidase activity. With M. voltae membranes as the enzyme source, signal peptidase activity in vitro was optimal between 30 and 40 degrees C; it was dependent on a low concentration of KCl or NaCl but was effective over a broad concentration range up to 1 M. Processing of the M. voltae S-layer protein at the predicted cleavage site (confirmed by N-terminal sequencing) was demonstrated with the overexpressed archaeal gene product. Although E. coli signal peptidase was able to correctly process the signal peptide during overexpression of the M. voltae S layer protein in vivo, the contribution of the E. coli signal peptidase to cleavage of the substrate in the in vitro assay was minimal since E. coli membranes alone did not show significant activity towards the S-layer substrate in in vitro assays. In addition, when the peptidase assays were performed in 1 M NaCl (a previously reported inhibitory condition for E. coli signal peptidase I), efficient processing of the substrate was observed only when the E. coli membranes contained overexpressed M. voltae signal peptidase. This is the first proof of expressed type I signal peptidase activity from a specific archaeal gene product. PMID- 14526004 TI - Osmotic adaptation of Thermus thermophilus RQ-1: lesson from a mutant deficient in synthesis of trehalose. AB - Strains of Thermus thermophilus accumulate primarily trehalose and smaller amounts of mannosylglycerate in response to salt stress in yeast extract containing media (O. C. Nunes, C. M. Manaia, M. S. da Costa, and H. Santos, Appl. Environ. Microbiol. 61:2351-2357, 1995). A 2.4-kbp DNA fragment from T. thermophilus strain RQ-1 carrying otsA (encoding trehalose-phosphate synthase [TPS]), otsB (encoding trehalose-phosphate phosphatase [TPP]), and a short sequence of the 5' end of treS (trehalose synthase [TreS]) was cloned from a gene library. The sequences of the three genes (including treS) were amplified by PCR and sequenced, revealing that the genes were structurally linked. To understand the role of trehalose during salt stress in T. thermophilus RQ-1, we constructed a mutant, designated RQ-1M6, in which TPS (otsA) and TPP (otsB) genes were disrupted by gene replacement. Mutant RQ-1M6 accumulated trehalose and mannosylglycerate in a medium containing yeast extract and NaCl. However, growth in a defined medium (without yeast extract, known to contain trehalose) containing NaCl led to the accumulation of mannosylglycerate but not trehalose. The deletion of otsA and otsB reduced the ability to grow in defined salt containing medium, with the maximum salinity being 5% NaCl for RQ-1 and 3% NaCl for RQ-1M6. The lower salt tolerance observed in the mutant was relieved by the addition of trehalose to the growth media. In contrast to trehalose, the addition of glycine betaine, mannosylglycerate, maltose, and glucose to the growth medium did not allow the mutant to grow at higher salinities. The results presented here provide crucial evidence for the importance of the TPS/TPP pathway for the synthesis and accumulation of trehalose and the decisive contribution of this disaccharide to osmotic adaptation in T. thermophilus RQ-1. PMID- 14526005 TI - Restricted translocation across the cell wall regulates secretion of the broad range phospholipase C of Listeria monocytogenes. AB - The virulence of Listeria monocytogenes is directly related to its ability to spread from cell to cell without leaving the intracellular milieu. During cell-to cell spread, bacteria become temporarily confined to secondary vacuoles. Among the bacterial factors involved in escape from these vacuoles is a secreted broad range phospholipase C (PC-PLC), the activation of which requires processing of an N-terminal prodomain. Mpl, a secreted metalloprotease of Listeria, is involved in the proteolytic activation of PC-PLC. We previously showed that, during intracellular growth, bacteria maintain a pool of PC-PLC that is not accessible to antibodies and that is rapidly released in its active form in response to a decrease in pH. pH-regulated release of active PC-PLC is Mpl dependent. To further characterize the mechanism regulating secretion of PC-PLC, the bacterial localization of PC-PLC and Mpl was investigated. Both proteins were detected in the bacterial supernatant and lysate with no apparent changes in molecular weight. Extraction of bacteria-associated PC-PLC and Mpl required cell wall hydrolysis, but there was no indication that either protein was covalently bound to the bacterial cell wall. Results from pulse-chase experiments performed with infected macrophages indicated that the rate of synthesis of PC-PLC exceeded the rate of translocation across the bacterial cell wall and confirmed that the pool of PC-PLC associated with bacteria was efficiently activated and secreted upon acidification of the host cell cytosol. These data suggest that bacterially associated PC-PLC and Mpl localize at the cell wall-membrane interface and that translocation of PC-PLC across the bacterial cell wall is rate limiting, resulting in the formation of a bacterially associated pool of PC-PLC that would readily be accessible for activation and release into nascent secondary vacuoles. PMID- 14526006 TI - An archaeal chromosomal autonomously replicating sequence element from an extreme halophile, Halobacterium sp. strain NRC-1. AB - We report on the identification and first cloning of an autonomously replicating sequence element from the chromosome of an archaeon, the extreme halophile Halobacterium strain NRC-1. The putative replication origin was identified by association with the orc7 gene and replication ability in the host strain, demonstrated by cloning into a nonreplicating plasmid. Deletion analysis showed that sequences located up to 750 bp upstream of the orc7 gene translational start, plus the orc7 gene and 50 bp downstream, are sufficient to endow the plasmid with replication ability, as judged by expression of a plasmid-encoded mevinolin resistance selectable marker and plasmid recovery after transformation. Sequences located proximal to the two other chromosomally carried haloarchaeal orc genes (orc6 and orc8) are not able to promote efficient autonomous replication. Located within the 750-bp region upstream of orc7 is a nearly perfect inverted repeat of 31 bp, which flanks an extremely AT-rich (44%) stretch of 189 bp. The replication ability of the plasmid was lost when one copy of the inverted repeat was deleted. Additionally, the inverted repeat structure near orc7 homologs in the genomic sequences of two other halophiles, Haloarcula marismortui and Haloferax volcanii, is highly conserved. Our results indicate that, in halophilic archaea, a chromosomal origin of replication is physically linked to orc7 homologs and that this element is sufficient to promote autonomous replication. We discuss the finding of a functional haloarchaeal origin in relation to the large number of orc1-cdc6 homologs identified in the genomes of all haloarchaea to date. PMID- 14526007 TI - The sloABCR operon of Streptococcus mutans encodes an Mn and Fe transport system required for endocarditis virulence and its Mn-dependent repressor. AB - Streptococcus mutans belongs to the viridans group of oral streptococci, which is the leading cause of endocarditis in humans. The LraI family of lipoproteins in viridans group streptococci and other bacteria have been shown to function as virulence factors, adhesins, or ABC-type metal transporters. We previously reported the identification of the S. mutans LraI operon, sloABCR, which encodes components of a putative metal uptake system composed of SloA, an ATP-binding protein, SloB, an integral membrane protein, and SloC, a solute-binding lipoprotein, as well as a metal-dependent regulator, SloR. We report here the functional analysis of this operon. By Western blotting, addition of Mn to the growth medium repressed SloC expression in a wild-type strain but not in a sloR mutant. Other metals tested had little effect. Cells were also tested for aerobic growth in media stripped of metals then reconstituted with Mg and either Mn or Fe. Fe at 10 micro M supported growth of the wild-type strain but not of a sloA or sloC mutant. Mn at 0.1 micro M supported growth of the wild-type strain and sloR mutant but not of sloA or sloC mutants. The combined results suggest that the SloABC proteins transport both metals, although the SloR protein represses this system only in response to Mn. These conclusions are supported by (55)Fe uptake studies with Mn as a competitor. Finally, a sloA mutant demonstrated loss of virulence in a rat model of endocarditis, suggesting that metal transport is required for endocarditis pathogenesis. PMID- 14526008 TI - Mechanism of Pseudomonas aeruginosa RhlR transcriptional regulation of the rhlAB promoter. AB - Pseudomonas aeruginosa contains two transcription regulators (LasR and RhlR) that, when complexed with their specific autoinducers (3-oxo-dodecanoyl homoserine lactone and butanoyl-homoserine lactone, respectively) activate transcription of different virulence-associated traits. We studied the RhlR dependent transcriptional regulation of the rhlAB operon encoding rhamnosyltransferase 1, an enzyme involved in the synthesis of the surfactant monorhamnolipid, and showed that RhlR binds to a specific sequence in the rhlAB regulatory region, both in the presence and in the absence of its autoinducer. Our data suggest that in the former case it activates transcription, whereas in the latter it acts as a transcriptional repressor of this promoter. RhlR seems to repress the transcription of other quorum-sensing-regulated genes; thus, RhlR repressor activity might be of importance in the finely regulated expression of P. aeruginosa virulence-associated traits. PMID- 14526009 TI - Constitutive activation of the Escherichia coli Pho regulon upregulates rpoS translation in an Hfq-dependent fashion. AB - Regulation of the sigma factor RpoS occurs at the levels of transcription, translation, and protein stability activity, and it determines whether Escherichia coli turns on or off the stationary-phase response. To better understand the regulation of RpoS, we conducted genetic screens and found that mutations in the pst locus cause accumulation of RpoS during exponential growth. The pst locus encodes for the components of the high-affinity transport system for inorganic phosphate (P(i)), which is involved in sensing P(i) levels in the environment. When the Pst transporter is compromised (either by mutation or by P(i) starvation), the two-component system PhoBR activates the transcription of the Pho regulon, a subset of genes that encode proteins for transporting and metabolizing alternative phosphate sources. Our data show that strains carrying mutations which constitutively activate the Pho regulon have increased rpoS translation during exponential growth. This upregulation of rpoS translation is Hfq dependent, suggesting the involvement of a small regulatory RNA (sRNA). The transcription of this yet-to-be-identified sRNA is regulated by the PhoBR two component system. PMID- 14526010 TI - Transcription regulation by tandem-bound FNR at Escherichia coli promoters. AB - FNR is an Escherichia coli transcription factor that regulates the transcription of many genes in response to anaerobiosis. We have constructed a series of artificial FNR-dependent promoters, based on the melR promoter, in which a consensus FNR binding site was centered at position -41.5 relative to the transcription start site. A second consensus FNR binding site was introduced at different upstream locations, and promoter activity was assayed in vivo. FNR can activate transcription from these promoters when the upstream FNR binding site is located at many different positions. However, sharp repression is observed when the upstream-bound FNR is located near positions -85 or -95. This repression is relieved by the FNR G74C substitution mutant, previously identified as being defective in transcription repression at the yfiD promoter. A parallel series of artificial FNR-dependent promoters, carrying a consensus FNR binding site at position -61.5 and a second upstream DNA site for FNR, was also constructed. Again, promoter activity was repressed by FNR when the upstream-bound FNR was located at particular positions. PMID- 14526011 TI - Characterization of the two Mycobacterium tuberculosis recA promoters. AB - The recA gene of Mycobacterium tuberculosis is unusual in that it is expressed from two promoters, one of which, P1, is DNA damage inducible independently of LexA and RecA, while the other, P2, is regulated by LexA in the classical way (E. O. Davis, B. Springer, K. K. Gopaul, K. G. Papavinasasundaram, P. Sander, and E. C. Bottger, Mol. Microbiol. 46:791-800, 2002). In this study we characterized these two promoters in more detail. Firstly, we localized the promoter elements for each of the promoters, and in so doing we identified a mutation in each promoter which eliminates promoter activity. Interestingly, a motif with similarity to Escherichia coli sigma(70) -35 elements but located much closer to the -10 element is important for optimal expression of P1, whereas the sequence at the -35 location is not. Secondly, we found that the sequences flanking the promoters can have a profound effect on the expression level directed by each of the promoters. Finally, we examined the contribution of each of the promoters to recA expression and compared their kinetics of induction following DNA damage. PMID- 14526012 TI - Rgg coordinates virulence factor synthesis and metabolism in Streptococcus pyogenes. AB - Streptococcus pyogenes is a human-specific pathogen that relies on its host for metabolic substrates. Rgg-like proteins constitute a family of transcriptional regulators present in several gram-positive bacteria. In S. pyogenes, Rgg influences the expression of several virulence-associated proteins localized to the cell wall and extracellular environment. Secreted enzymes may degrade host macromolecules, thereby liberating metabolic substrates. To determine if Rgg regulation of exoprotein expression is associated with altered metabolism, the catabolic activities of S. pyogenes strain NZ131 (serotype M49) and an isogenic rgg mutant strain were analyzed during growth with complex and defined media. As expected, the wild-type strain preferentially used glucose and produced lactic acid during the exponential phase of growth. In contrast, the rgg mutant fermented arginine in the exponential phase of growth, even in the presence of glucose. Arginine degradation was associated with a neutral culture pH and excretion of NH(3) and ornithine. Arginine, serine, and asparagine were depleted from mutant cultures during growth. The addition of arginine and serine to culture media increased the growth yield and NH(3) production of mutant but not wild-type cultures. Addition of asparagine had no effect on the growth yield of either strain. Altered metabolism of arginine and serine in the mutant was associated with increased transcript levels of genes encoding arginine deiminase and a putative serine dehydratase. Thus, Rgg coordinates virulence factor synthesis and catabolic activity and may be important in the pathogen's adaptation to changes in the availability of metabolic substrates. PMID- 14526014 TI - The iron-responsive regulator fur is transcriptionally autoregulated and not essential in Neisseria meningitidis. AB - Fur is a well-known iron-responsive repressor of gene transcription, which is used by many bacteria to respond to the low-iron environment that pathogens encounter during infection. The fur gene in Neisseria meningitidis has been described as an essential gene that may regulate a broad array of genes. We succeeded in obtaining an N. meningitidis mutant with the fur gene knocked out and used it to undertake studies of fur-mediated iron regulation. We show that expression of both Fur and the transferrin binding protein Tbp2 is iron regulated and demonstrate that this regulation is Fur mediated for the Tbp2 protein. Footprinting analysis revealed that Fur binds to two distinct sites upstream of its coding region with different affinities and that these binding sites overlap two promoters that differentially control transcription of the fur gene in response to iron. The presence of two independently regulated fur promoters may allow meningococcus to fine-tune expression of this regulator controlling iron homeostasis, possibly during infection. PMID- 14526013 TI - DpiA binding to the replication origin of Escherichia coli plasmids and chromosomes destabilizes plasmid inheritance and induces the bacterial SOS response. AB - The dpiA and dpiB genes of Escherichia coli, which are orthologs of genes that regulate citrate uptake and utilization in Klebsiella pneumoniae, comprise a two component signal transduction system that can modulate the replication of and destabilize the inheritance of pSC101 and certain other plasmids. Here we show that perturbed replication and inheritance result from binding of the effector protein DpiA to A+T-rich replication origin sequences that resemble those in the K. pneumoniae promoter region targeted by the DpiA ortholog, CitB. Consistent with its ability to bind to A+T-rich origin sequences, overproduction of DpiA induced the SOS response in E. coli, suggesting that chromosomal DNA replication is affected. Bacteria that overexpressed DpiA showed an increased amount of DNA per cell and increased cell size-both also characteristic of the SOS response. Concurrent overexpression of the DNA replication initiation protein, DnaA, or the DNA helicase, DnaB-both of which act at A+T-rich replication origin sequences in the E. coli chromosome and DpiA-targeted plasmids-reversed SOS induction as well as plasmid destabilization by DpiA. Our finding that physical and functional interactions between DpiA and sites of replication initiation modulate DNA replication and plasmid inheritance suggests a mechanism by which environmental stimuli transmitted by these gene products can regulate chromosomal and plasmid dynamics. PMID- 14526015 TI - Regulation of expression of the 2-deoxy-D-ribose utilization regulon, deoQKPX, from Salmonella enterica serovar typhimurium. AB - Salmonella enterica, in contrast to Escherichia coli K12, can use 2-deoxy-D ribose as the sole carbon source. The genetic determinants for this capacity in S. enterica serovar Typhimurium include four genes, of which three, deoK, deoP, and deoX, constitute an operon. The fourth, deoQ, is transcribed in the opposite direction. The deoK gene encodes deoxyribokinase. In silico analyses indicated that deoP encodes a permease and deoQ encodes a regulatory protein of the deoR family. The deoX gene product showed no match to known proteins in the databases. Deletion analyses showed that both a functional deoP gene and a functional deoX gene were required for optimal utilization of deoxyribose. Using gene fusion technology, we observed that deoQ and the deoKPX operon were transcribed from divergent promoters located in the 324-bp intercistronic region between deoQ and deoK. The deoKPX promoter was 10-fold stronger than the deoQ promoter, and expression was negatively regulated by DeoQ as well as by DeoR, the repressor of the deoxynucleoside catabolism operon. Transcription of deoKPX but not of deoQ was regulated by catabolite repression. Primer extension analysis identified the transcriptional start points of both promoters and showed that induction by deoxyribose occurred at the level of transcription initiation. Gel retardation experiments with purified DeoQ illustrated that it binds independently to tandem operator sites within the deoQ and deoK promoter regions with K(d) values of 54 and 2.4 nM, respectively. PMID- 14526016 TI - A second PDZ-containing serine protease contributes to activation of the sporulation transcription factor sigmaK in Bacillus subtilis. AB - Gene expression late during the process of sporulation in Bacillus subtilis is governed by a multistep, signal transduction pathway involving the transcription factor sigma(K), which is derived by regulated proteolysis from the inactive proprotein pro-sigma(K). Processing of pro-sigma(K) is triggered by a signaling protein known as SpoIVB, a serine protease that contains a region with similarity to the PDZ family of protein-protein interaction domains. Here we report the discovery of a second PDZ-containing serine protease called CtpB that contributes to the activation of the pro-sigma(K) processing pathway. CtpB is a sporulation specific, carboxyl-terminal processing protease and shares several features with SpoIVB. We propose that CtpB acts to fine-tune the regulation of pro-sigma(K) processing, and we discuss possible models by which CtpB influences the sigma(K) activation pathway. PMID- 14526018 TI - Regulation of expression of cellulosomal cellulase and hemicellulase genes in Clostridium cellulovorans. AB - The regulation of expression of the genes encoding the cellulases and hemicellulases of Clostridium cellulovorans was studied at the mRNA level with cells grown under various culture conditions. A basic pattern of gene expression and of relative expression levels was obtained from cells grown in media containing poly-, di- or monomeric sugars. The cellulase (cbpA and engE) and hemicellulase (xynA) genes were coordinately expressed in medium containing cellobiose or cellulose. Growth in the presence of cellulose, xylan, and pectin gave rise to abundant expression of most genes (cbpA-exgS, engH, hbpA, manA, engM, engE, xynA, and/or pelA) studied. Moderate expression of cbpA, engH, manA, engE, and xynA was observed when cellobiose or fructose was used as the carbon source. Low levels of mRNA from cbpA, manA, engE, and xynA were observed with cells grown in lactose, mannose, and locust bean gum, and very little or no expression of cbpA, engH, manA, engE, and xynA was detected in glucose-, galactose-, maltose-, and sucrose-grown cells. The cbpA-exgS and engE genes were most frequently expressed under all conditions studied, whereas expression of xynA and pelA was more specifically induced at higher levels in xylan- or pectin containing medium, respectively. Expression of the genes (cbpA, hbpA, manA, engM, and engE) was not observed in the presence of most soluble di- or monosaccharides such as glucose. These results support the hypotheses that there is coordinate expression of some cellulases and hemicellulases, that a catabolite repression type of mechanism regulates cellulase expression in rapidly growing cells, and that the presence of hemicelluloses has an effect on cellulose utilization by the cell. PMID- 14526017 TI - Positive correlation between tyrosine phosphorylation of CpsD and capsular polysaccharide production in Streptococcus pneumoniae. AB - CpsA, CpsB, CpsC, and CpsD are part of a tyrosine phosphorylation regulatory system involved in modulation of capsule synthesis in Streptococcus pneumoniae and many other gram-positive and gram-negative bacteria. Using an immunoblotting technique, we observed distinct laddering patterns of S. pneumoniae capsular polysaccharides of various serotypes and found that transfer of the polymer from the membrane to the cell wall was independent of size. Deletion of cps2A, cps2B, cps2C, or cps2D in the serotype 2 strain D39 did not affect the ability to transfer capsule to the cell wall. Deletion of cps2C or cps2D, which encode two domains of an autophosphorylating tyrosine kinase, resulted in the production of only short-chain polymers. The function of Cps2A is unknown, and the polymer laddering pattern of the cps2A deletion mutants appeared similar to that of the parent, although the total amount of capsule was decreased. Loss of Cps2B, a tyrosine phosphatase and a kinase inhibitor, resulted in an increase in capsule amount and a normal ladder pattern. However, Cps2B mutants exhibited reduced virulence following intravenous inoculation of mice and were unable to colonize the nasopharynx, suggesting a diminished capacity to sense or respond to these environments. In D39 and its isogenic mutants, the amounts of capsule and tyrosine-phosphorylated Cps2D (Cps2D approximately P) correlated directly. In contrast, restoration of type 2 capsule production followed by deletion of cps2B in Rx1, a laboratory passaged D39 derivative containing multiple uncharacterized mutations, resulted in decreased capsule amounts but no alteration in Cps2D approximately P levels. Thus, a factor outside the capsule locus, which is either missing or defective in the Rx1 background, is important in the control of capsule synthesis. PMID- 14526019 TI - Spontaneously arising mutL mutators in evolving Escherichia coli populations are the result of changes in repeat length. AB - Over the course of thousands of generations of growth in a glucose-limited environment, 3 of 12 experimental populations of Escherichia coli spontaneously and independently evolved greatly increased mutation rates. In two of the populations, the mutations responsible for this increased mutation rate lie in the same region of the mismatch repair gene mutL. In this region, a 6-bp repeat is present in three copies in the gene of the wild-type ancestor of the experimental populations but is present in four copies in one of the experimental populations and two copies in the other. These in-frame mutations either add or delete the amino acid sequence LA in the MutL protein. We determined that the replacement of the wild-type sequence with either of these mutations was sufficient to increase the mutation rate of the wild-type strain to a level comparable to that of the mutator strains. Complementation of strains bearing the mutator mutations with wild-type copies of either mutL or the mismatch repair gene uvrD rescued the wild-type mutation rate. The position of the mutator mutations-in the region of MutL known as the ATP lid-suggests a possible deficiency in MutL's ATPase activity as the cause of the mutator phenotype. The similarity of the two mutator mutations (despite the independent evolutionary histories of the populations that gave rise to them) leads to a discussion of the potential adaptive role of DNA repeats. PMID- 14526021 TI - Effects of carboxy-terminal modifications and pH on binding of a Bacillus subtilis small, acid-soluble spore protein to DNA. AB - Variants of the wild-type Bacillus subtilis alpha/beta-type small, acid-soluble spore protein (SASP) SspC(wt) were designed to evaluate the contribution of C terminal residues to these proteins' affinity for DNA. SspC variants lacking one to three C-terminal residues were similar to SspC(wt) in DNA binding, but removal of six C-terminal residues greatly decreased DNA binding. In contrast, a C terminal extension of three residues increased SspC's affinity for DNA 5- to 10 fold. C-terminal and N-terminal changes that independently caused large increases in SspC-DNA binding affinity were combined and produced an additive effect on DNA binding; the affinity of the resulting variant, SspC(DeltaN11-D13K-C3), for DNA was increased >/==" BORDER="0">20-fold over that of SspC(wt). For most of the SspC variants tested, lowering the pH from 7 to 6 improved DNA binding two- to sixfold, although the opposite effect was observed with variants having additional C-terminal basic residues. In vitro, the binding of SspC(DeltaN11-D13K C3) to DNA suppressed the formation of cyclobutane-type thymine dimers and promoted the formation of the spore photoproduct upon UV irradiation to the same degree as the binding of SspC(wt). However, B. subtilis spores lacking major alpha/beta-type SASP and overexpressing SspC(DeltaN11-D13K-C3) had a 10-fold lower viability and far less UV and heat resistance than spores overexpressing SspC(wt). This apparent lack of DNA protection by SspC(DeltaN11-D13K-C3) in vivo is likely due to the twofold-lower level of this protein in spores compared to the level of SspC(wt), perhaps because of effects of SspC(DeltaN11-D13K-C3) on gene expression in the forespore during sporulation. The latter results indicate that only moderately strong binding of alpha/beta-type SASP to DNA is important to balance the potentially conflicting requirements for these proteins in DNA transcription and DNA protection during spore formation, spore dormancy, and spore germination and outgrowth. PMID- 14526020 TI - Global mutational analysis of NtrC-like activators in Myxococcus xanthus: identifying activator mutants defective for motility and fruiting body development. AB - The multicellular developmental cycle of Myxococcus xanthus requires large-scale changes in gene transcription, and recent findings indicate that NtrC-like activators play a prominent role in regulating these changes. In this study, we made insertions in 28 uncharacterized ntrC-like activator (nla) genes and found that eight of these insertions cause developmental defects. Hence, these results are consistent with the idea that M. xanthus uses a series of different NtrC-like activators during fruiting body development. Four of the eight developmental mutants we identified have motility defects. The nla1, nla19, and nla23 mutants show S-motility defects, while the nla24 mutant shows defects in both S-motility and A-motility. During development, aggregation of the nla1, nla19, and nla23 mutants is delayed slightly and the nla24 mutant shows no signs of aggregation or sporulation. The nla4, nla6, nla18, and nla28 mutants have no appreciable loss in motility, but they fail to aggregate and to sporulate normally. The nla18 mutant belongs to a special class of developmental mutants whose defects can be rescued when they are codeveloped with wild-type cells, suggesting that nla18 fails to produce a cell-cell signal required for development. The three remaining activator mutants, nla4, nla6, and nla28, appear to have complex developmental phenotypes that include deficiencies in cell-cell developmental signals. PMID- 14526022 TI - Analysis of the mobilization region of the broad-host-range IncQ-like plasmid pTC F14 and its ability to interact with a related plasmid, pTF-FC2. AB - Plasmid pTC-F14 is a 14.2-kb plasmid isolated from Acidithiobacillus caldus that has a replicon that is closely related to the promiscuous, broad-host-range IncQ family of plasmids. The region containing the mobilization genes was sequenced and encoded five Mob proteins that were related to those of the DNA processing (Dtr or Tra1) region of IncP plasmids rather than to the three-Mob-protein system of the IncQ group 1 plasmids (e.g., plasmid RSF1010 or R1162). Plasmid pTC-F14 is the second example of an IncQ family plasmid that has five mob genes, the other being pTF-FC2. The minimal region that was essential for mobilization included the mobA, mobB, and mobC genes, as well as the oriT gene. The mobD and mobE genes were nonessential, but together, they enhanced the mobilization frequency by approximately 300-fold. Mobilization of pTC-F14 between Escherichia coli strains by a chromosomally integrated RP4 plasmid was more than 3,500-fold less efficient than the mobilization of pTF-FC2. When both plasmids were coresident in the same E. coli host, pTC-F14 was mobilized at almost the same frequency as pTF-FC2. This enhanced pTC-F14 mobilization frequency was due to the presence of a combination of the pTF-FC2 mobD and mobE gene products, the functions of which are still unknown. Mob protein interaction at the oriT regions was unidirectionally plasmid specific in that a plasmid with the oriT region of pTC-F14 could be mobilized by pTF-FC2 but not vice versa. No evidence for any negative effect on the transfer of one plasmid by the related, potentially competitive plasmid was obtained. PMID- 14526023 TI - Cryo-transmission electron microscopy of frozen-hydrated sections of Escherichia coli and Pseudomonas aeruginosa. AB - High-pressure freezing of Escherichia coli K-12 and Pseudomonas aeruginosa PAO1 in the presence of cryoprotectants provided consistent vitrification of cells so that frozen-hydrated sections could be cut, providing approximately 2-nm resolution of structure. The size and shape of the bacteria, as well as their surface and cytoplasmic constituents, were nicely preserved and compared well with other published high-resolution techniques. Cells possessed a rich cytoplasm containing a diffuse dispersion of ribosomes and genetic material. Close examination of cells revealed that the periplasmic space was compressed during cryosectioning, a finding which provided supporting evidence that this space is filled by a compressible gel. Since the outer membrane and peptidoglycan layer are bonded together via lipoproteins, the space between them (although still part of the periplasmic space) was not as compacted. Even when this cryosectioning compression was taken into account, there was still substantial variability in the width of the periplasmic space. It is possible that the protoplast has some capacity to float freely within the periplasm. PMID- 14526024 TI - 2-Oxoglutarate:NADP(+) oxidoreductase in Azoarcus evansii: properties and function in electron transfer reactions in aromatic ring reduction. AB - The conversion of [(14)C]benzoyl-coenzyme A (CoA) to nonaromatic products in the denitrifying beta-proteobacterium Azoarcus evansii grown anaerobically on benzoate was investigated. With cell extracts and 2-oxoglutarate as the electron donor, benzoyl-CoA reduction occurred at a rate of 10 to 15 nmol min(-1) mg(-1). 2-Oxoglutarate could be replaced by dithionite (200% rate) and by NADPH ( approximately 10% rate); in contrast NADH did not serve as an electron donor. Anaerobic growth on aromatic compounds induced 2-oxoglutarate:acceptor oxidoreductase (KGOR), which specifically reduced NADP(+), and NADPH:acceptor oxidoreductase. KGOR was purified by a 76-fold enrichment. The enzyme had a molecular mass of 290 +/- 20 kDa and was composed of three subunits of 63 (gamma), 62 (alpha), and 37 (beta) kDa in a 1:1:1 ratio, suggesting an (alphabetagamma)(2) composition. The native enzyme contained Fe (24 mol/mol of enzyme), S (23 mol/mol), flavin adenine dinucleotide (FAD; 1.4 mol/mol), and thiamine diphosphate (0.95 mol/mol). KGOR from A. evansii was highly specific for 2-oxoglutarate as the electron donor and accepted both NADP(+) and oxidized viologens as electron acceptors; in contrast NAD(+) was not reduced. These results suggest that benzoyl-CoA reduction is coupled to the complete oxidation of the intermediate acetyl-CoA in the tricarboxylic acid cycle. Electrons generated by KGOR can be transferred to both oxidized ferredoxin and NADP(+), depending on the cellular needs. N-terminal amino acid sequence analysis revealed that the open reading frames for the three subunits of KGOR are similar to three adjacently located open reading frames in Bradyrhizobium japonicum. We suggest that these genes code for a very similar three-subunit KGOR, which may play a role in nitrogen fixation. The alpha-subunit is supposed to harbor one FAD molecule, two [4Fe-4S] clusters, and the NADPH binding site; the beta-subunit is supposed to harbor one thiamine diphosphate molecule and one further [4Fe-4S] cluster; and the gamma-subunit is supposed to harbor the CoA binding site. This is the first study of an NADP(+)-specific KGOR. A similar NADP(+)-specific pyruvate oxidoreductase, which contains all domains in one large subunit, has been reported for the mitochondrion of the protist Euglena gracilis and the apicomplexan Cryptosporidium parvum. PMID- 14526025 TI - Molecular characterization of Brucella abortus chromosome II recombination. AB - Large-scale genomic rearrangements including inversions, deletions, and duplications are significant in bacterial evolution. The recently completed Brucella melitensis 16M and Brucella suis 1330 genomes have facilitated the investigation of such events in the Brucella spp. Suppressive subtractive hybridization (SSH) was employed in identifying genomic differences between B. melitensis 16M and Brucella abortus 2308. Analysis of 45 SSH clones revealed several deletions on chromosomes of B. abortus and B. melitensis that encoded proteins of various metabolic pathways. A 640-kb inversion on chromosome II of B. abortus has been reported previously (S. Michaux Charachon, G. Bourg, E. Jumas Bilak, P. Guigue Talet, A. Allardet Servent, D. O'Callaghan, and M. Ramuz, J. Bacteriol. 179:3244-3249, 1997) and is further described in this study. One end of the inverted region is located on a deleted TATGC site between open reading frames BMEII0292 and BMEII0293. The other end inserted at a GTGTC site of the cyclic-di-GMP phosphodiesterase A (PDEA) gene (BMEII1009), dividing PDEA into two unequal DNA segments of 160 and 977 bp. As a consequence of inversion, the 160-bp segment that encodes the N-terminal region of PDEA was relocated at the opposite end of the inverted chromosomal region. The splitting of the PDEA gene most likely inactivated the function of this enzyme. A recombination mechanism responsible for this inversion is proposed. PMID- 14526026 TI - ATP-bound conformation of topoisomerase IV: a possible target for quinolones in Streptococcus pneumoniae. AB - Topoisomerase IV, a C(2)E(2) tetramer, is involved in the topological changes of DNA during replication. This enzyme is the target of antibacterial compounds, such as the coumarins, which target the ATP binding site in the ParE subunit, and the quinolones, which bind, outside the active site, to the quinolone resistance determining region (QRDR). After site-directed and random mutagenesis, we found some mutations in the ATP binding site of ParE near the dimeric interface and outside the QRDR that conferred quinolone resistance to Streptococcus pneumoniae, a bacterial pathogen. Modeling of the N-terminal, 43-kDa ParE domain of S. pneumoniae revealed that the most frequent mutations affected conserved residues, among them His43 and His103, which are involved in the hydrogen bond network supporting ATP hydrolysis, and Met31, at the dimeric interface. All mutants showed a particular phenotype of resistance to fluoroquinolones and an increase in susceptibility to novobiocin. All mutations in ParE resulted in resistance only when associated with a mutation in the QRDR of the GyrA subunit. Our models of the closed and open conformations of the active site indicate that quinolones preferentially target topoisomerase IV of S. pneumoniae in its ATP-bound closed conformation. PMID- 14526027 TI - Identification of three new genes involved in morphogenesis and antibiotic production in Streptomyces coelicolor. AB - We report the isolation and partial characterization of three new mutants of Streptomyces coelicolor that are defective in morphogenesis and antibiotic production. The genes identified by the mutations were located and cloned by using a combination of Tn5 in vitro mutagenesis, cotransformation, and genetic complementation. Mutant SE69 produces lower amounts of antibiotics than the wild type produces, produces spores only after prolonged incubation on rich media, and identifies a gene whose predicted protein product is similar to the GntR family of transcriptional regulators; also, production of aerial mycelia on both rich and poor media is significantly delayed in this mutant. Mutant SE293 is defective in morphogenesis, overproduces antibiotics on rich media, fails to grow on minimal media, and identifies a gene whose predicted protein product is similar to the TetR family of transcriptional regulators. Preliminary evidence suggests that the SE293 gene product may control a molybdopterin binding protein located immediately adjacent to it. Mutant SJ175 sporulates sooner and more abundantly than the wild type and overproduces antibiotics on rich media, and it identifies a gene whose predicted protein product contains regions of predominantly hydrophobic residues similar to those of integral membrane proteins. PMID- 14526028 TI - Interfering with different steps of protein synthesis explored by transcriptional profiling of Escherichia coli K-12. AB - Escherichia coli responses to four inhibitors that interfere with translation were monitored at the transcriptional level. A DNA microarray method provided a comprehensive view of changes in mRNA levels after exposure to these agents. Real time reverse transcriptase PCRanalysis served to verify observations made with microarrays, and a chromosomal grpE::lux operon fusion was employed to specifically monitor the heat shock response. 4-Azaleucine, a competitive inhibitor of leucyl-tRNA synthetase, surprisingly triggered the heat shock response. Administration of mupirocin, an inhibitor of isoleucyl-tRNA synthetase activity, resulted in changes reminiscent of the stringent response. Treatment with kasugamycin and puromycin (targeting ribosomal subunit association as well as its peptidyl-transferase activity) caused accumulation of mRNAs from ribosomal protein operons. Abundant biosynthetic transcripts were often significantly diminished after treatment with any of these agents. Exposure of a relA strain to mupirocin resulted in accumulation of ribosomal protein operon transcripts. However, the relA strain's response to the other inhibitors was quite similar to that of the wild-type strain. PMID- 14526029 TI - A second and unusual pucBA operon of Rhodobacter sphaeroides 2.4.1: genetics and function of the encoded polypeptides. AB - A new operon (designated the puc2BA operon) displaying a high degree of similarity to the original pucBA genes of Rhodobacter sphaeroides 2.4.1 (designated puc1) was identified and studied genetically and biochemically. The puc2B-encoded polypeptide is predicted to exhibit 94% identity with the original beta-apoprotein. The puc2A-encoded polypeptide is predicted to be much larger (263 amino acids) than the 54-amino-acid puc1A-encoded polypeptide. In the first 48 amino acids of the puc2A-encoded polypeptide there is 58% amino acid sequence identity to the original puc1A-encoded polypeptide. We found that puc2BA is expressed, and DNA sequence data suggested that puc2BA is regulated by the PpsR/AppA repressor-antirepressor and FnrL. Employing genetic and biochemical approaches, we obtained evidence that the puc2B-encoded polypeptide is able to enter into LH2 complex formation, but neither the full-length puc2A-encoded polypeptide nor its N-terminal 48-amino-acid derivative is able to enter into LH2 complex formation. Thus, the sole source of alpha-polypeptides for the LH2 complex is puc1A. The role of the puc1C-encoded polypeptide was also determined. We found that the presence of this polypeptide is essential for normal levels of transcription and translation of the puc1 operon but not for transcription and translation of the puc2 operon. Thus, the puc1C gene product appears to have both transcriptional and posttranscriptional roles in LH2 formation. Finally, the absence of any LH2 complex when puc1B was deleted in frame was surprising since we know that in the presence of functional puc2BA, approximately 30% of the LH2 complexes normally observed contain a puc2B-encoded beta-polypeptide. PMID- 14526030 TI - Changes in Escherichia coli rRNA promoter activity correlate with changes in initiating nucleoside triphosphate and guanosine 5' diphosphate 3'-diphosphate concentrations after induction of feedback control of ribosome synthesis. AB - rRNA synthesis is the rate-limiting step in ribosome synthesis in Escherichia coli. Its regulation has been described in terms of a negative-feedback control loop in which rRNA promoter activity responds to the amount of translation. The feedback nature of this control system was demonstrated previously by artificially changing ribosome synthesis rates and observing responses of rRNA promoters. However, it has not been demonstrated previously that the initiating nucleoside triphosphate (iNTP) and guanosine 5'-diphosphate 3'-diphosphate (ppGpp), the molecular effectors responsible for controlling rRNA promoters in response to changes in the nutritional environment, are responsible for altering rRNA promoter activities under these feedback conditions. Here, we show that most feedback situations result in changes in the concentrations of both the iNTP and ppGpp and that the directions of these changes are consistent with a role for these two small-molecule regulators in feedback control of rRNA synthesis. In contrast, we observed no change in the level of DNA supercoiling under the feedback conditions examined. PMID- 14526031 TI - Changes in the concentrations of guanosine 5'-diphosphate 3'-diphosphate and the initiating nucleoside triphosphate account for inhibition of rRNA transcription in fructose-1,6-diphosphate aldolase (fda) mutants. AB - Early screens for conditional lethal mutations that affected rRNA expression in Escherichia coli identified temperature-sensitive fda mutants (fda encodes the glycolytic enzyme fructose-1,6-diphosphate aldolase). It was shown that these fda(Ts) mutants were severely impaired in rRNA synthesis upon shift to the restrictive temperature, although the mechanism of inhibition was never determined. Here, we bring resolution to this long-standing question by showing that changes in the concentrations of guanosine 5'-diphosphate 3'-diphosphate and initiating nucleoside triphosphates can account for the previously observed effects of fda mutations on rRNA transcription. PMID- 14526032 TI - Mutations affecting DNA-binding activity of the MexR repressor of mexR-mexA-mexB oprM operon expression. AB - We have isolated 25 MexR mutants that retained their dimerizing ability but were unable to bind mexOP DNA. Surprisingly, 20 mutations were located in the hydrophobic core region at alpha4, W1, alpha2, alpha3, and beta2, and only 3 were in positively charged residues. These results verified that DNA binding is mediated by distinct regions of MexR and showed the importance of the hydrophobic core region of the DNA-binding domain. PMID- 14526033 TI - Reduced water availability influences the dynamics, development, and ultrastructural properties of Pseudomonas putida biofilms. AB - Pseudomonas putida strain mt-2 unsaturated biofilm formation proceeds through three distinct developmental phases, culminating in the formation of a microcolony. The form and severity of reduced water availability alter cell morphology, which influences microcolony size and ultrastructure. The dehydration (matric stress) treatments resulted in biofilms comprised of smaller cells, but they were taller and more porous and had a thicker extracellular polysaccharide layer at the air interface. In the solute stress treatments, cell filamentation occurred more frequently in the presence of high concentrations of ionic (but not nonionic) solutes, and these filamented cells drastically altered the biofilm architecture. PMID- 14526034 TI - The SKHR motif is required for biological function of the VirR response regulator from Clostridium perfringens. AB - The response regulator VirR and its cognate sensor histidine kinase, VirS, are responsible for toxin gene regulation in the human pathogen Clostridium perfringens. The C-terminal domain of VirR (VirRc) contains the functional FxRxHrS motif, which is involved in DNA binding and is conserved in many regulatory proteins. VirRc was cloned, purified, and shown by in vivo and in vitro studies to comprise an independent DNA binding domain. Random and site directed mutagenesis was used to identify further amino acids that were required for the functional integrity of the protein. Random mutagenesis identified a unique residue, Met-172, that was required for biological function. Site-directed mutagenesis of the SKHR motif (amino acids 216 to 219) revealed that these residues were also required for biological activity. Analysis of the mutated proteins indicated that they were unable to bind to the DNA target with the same efficiency as the wild-type protein. PMID- 14526035 TI - Characterization of divIVA and other genes located in the chromosomal region downstream of the dcw cluster in Streptococcus pneumoniae. AB - We analyzed the chromosome region of Streptococcus pneumoniae located downstream of the division and cell wall (dcw) cluster that contains the homolog of the Bacillus subtilis cell division gene divIVA and some genes of unknown function. Inactivation of divIVA in S. pneumoniae resulted in severe growth inhibition and defects in cell shape, nucleoid segregation, and cell division. Inactivation of the ylm genes resulted in some morphological and/or division abnormalities, depending on the inactivated gene. Transcriptional analysis revealed a relationship between these genes and the ftsA and ftsZ cell division genes, also indicating that the connection between the dcw cluster and the divIVA region is more extensive than just chromosomal position and gene organization. PMID- 14526036 TI - Nucleotide-dependent conformational changes in the sigma54-dependent activator DctD. AB - Activators of sigma(54)-RNA polymerase holoenzyme couple ATP hydrolysis to formation of an open promoter complex. DctD(Delta1-142), a truncated and constitutively active form of the sigma(54)-dependent activator DctD from Sinorhizobium meliloti, displayed an altered DNase I footprint at its binding site located upstream of the dctA promoter in the presence of ATP. The altered footprint was not observed for a mutant protein with a substitution at or near the putative arginine finger, a conserved arginine residue thought to contact the nucleotide. These data suggest that structural changes in DctD(Delta1-142) during ATP hydrolysis can be detected by alterations in the DNase I footprint of the protein and may be communicated by interactions between bound nucleotide and the arginine finger. In addition, kinetic data for changes in fluorescence energy transfer upon binding of 2'(3')-O-(N-methylanthraniloyl)-ATP (Mant-ATP) to DctD(Delta1-142) and DctD suggested that these proteins undergo multiple conformational changes following ATP binding. PMID- 14526037 TI - Metagenomic analyses of an uncultured viral community from human feces. AB - Here we present the first metagenomic analyses of an uncultured viral community from human feces, using partial shotgun sequencing. Most of the sequences were unrelated to anything previously reported. The recognizable viruses were mostly siphophages, and the community contained an estimated 1,200 viral genotypes. PMID- 14526038 TI - Risk of ischemic stroke in patients with symptomatic vertebrobasilar stenosis undergoing surgical procedures. AB - BACKGROUND AND PURPOSE: There is little information to provide an estimate for stroke risk in patients with established stenosis or occlusion in the basilar or intracranial vertebral arteries undergoing surgical procedures. The objective of this study was to determine the ischemic stroke risk in this specific patient population. METHODS: A medical records linkage system retrospectively identified patients with a diagnosis of symptomatic vertebrobasilar stenosis or occlusion matched with surgical procedures. Patients were selected if they had stenosis or occlusion of the basilar or intracranial vertebral arteries identified on vascular imaging before undergoing surgical procedures under general anesthesia. Clinical and radiographic features were reviewed, along with the nature of the surgeries and details of the perioperative management. Records were reviewed for the diagnosis of stroke occurring within 1 month of surgery. RESULTS: Thirty eight patients with a history of symptomatic vertebrobasilar ischemia underwent 50 operations under general anesthesia, and 3 had ischemic strokes in the vertebrobasilar territory immediately after surgery, a per-procedure rate of 6.0% (95% confidence interval, 1.2 to 16.6) All 3 had episodes of prolonged hypotension (systolic blood pressure <100 mm Hg for >10 minutes) during surgery. CONCLUSIONS: The risk of perioperative stroke in patients with vertebrobasilar stenosis undergoing surgery under general anesthesia is 6.0%, which is notably higher than the risk for patients with other patterns of cerebrovascular disease. PMID- 14526039 TI - Wall mechanics of the stented extracranial carotid artery. AB - BACKGROUND AND PURPOSE: Abrupt compliance changes and concomitant nonlaminar flow patterns may contribute to endothelial dysfunction and subsequent neointimal thickening. The aim of this study was to test the feasibility of wall mechanics measurement using B-mode ultrasound image analysis by dedicated software in the stented human carotid artery. METHODS: Carotid Wallstents (Schneider) were placed in the extracranial carotid arteries of 15 patients. B-mode ultrasound examination was performed with a 7.5-MHz probe on the carotid artery upstream; at the proximal, mid, and distal stent levels; downstream from the stent; and on the contralateral internal and common carotid arteries. Carotid diameter (d) and systolic diameter changes (Deltad) were measured with a dedicated image processing system (IO version 3.1, IODP), while pulse blood pressure (DeltaP) was measured. Diameter compliance (Cd) and distensibility coefficient (DC) were calculated as Cd=2Deltad/DeltaP and DC=2Deltad/DeltaP/d and compared between measurement sites. RESULTS: The evaluation could be completed in 8 of 15 patients. Compliance was significantly lower at the proximal, mid, and distal stent levels (27.77+/-1.11, 27.38+/-1.08, 27.38+/-1.09x10(-3) mm x kPa(-1)) than upstream (103.3+/-36.7x10(-3) mm x kPa(-1)), downstream (91.5+/-41.3x10(-3) mm x kPa(-1)), or on the contralateral internal (87.6+/-28x10(-3) mm x kPa(-1)) and common (149.3+/-47.6x10(-3) mm x kPa(-1)) carotid arteries. CONCLUSIONS: Stenting of the extracranial carotid artery induces a compliance mismatch between the native and the stented artery. PMID- 14526040 TI - Risk factors for death or stroke after carotid endarterectomy: observations from the Ontario Carotid Endarterectomy Registry. AB - BACKGROUND AND PURPOSE: Carotid endarterectomy is an effective method for preventing strokes if patients do not suffer adverse perioperative outcomes. The purpose of this study was to identify preoperative patient risk factors for adverse outcomes (death or nonfatal stroke) after carotid endarterectomy through the use of a large population-based registry from Ontario, Canada. METHODS: Medical records of all 6038 patients who underwent carotid endarterectomy in Ontario between January 1, 1994, and December 31, 1997, were abstracted from 34 hospitals. Patient characteristics (demographic data, past medical history, neurological symptoms, comorbidities, radiological findings) and 30-day postoperative death or stroke rates were analyzed with logistic regression analysis. RESULTS: The overall 30-day death or stroke rate after surgery was 6.0%. A history of transient ischemic attack or stroke (odds ratio [OR], 1.75; 95% confidence interval [CI], 1.39 to 2.20), atrial fibrillation (OR, 1.89; 95% CI, 1.29 to 2.76), contralateral carotid occlusion (OR, 1.72; 95% C.I., 1.25 to 2.38), congestive heart failure (OR, 1.80; 95% CI, 1.15 to 2.81), and diabetes (OR, 1.28; 95% CI, 1.01 to 1.63) were significant independent predictors for 30 day death or stroke. These 5 factors were combined into a simple risk score that can be used to stratify patients into different risk groups for complications after surgery. CONCLUSIONS: Several patient characteristics predict the development of stroke and death after carotid endarterectomy. These characteristics may help clinicians in patient counseling and contribute to studies "benchmarking" the outcomes of carotid surgery in the community setting. PMID- 14526041 TI - Editorial comment--Identifying risk factors for perioperative outcomes after carotid endarterectomy: the story continues. PMID- 14526042 TI - Biomedical research. Speeding up delivery: NIH aims to push for clinical results. PMID- 14526043 TI - European Research Council. Experts put some flesh on a bare-bones proposal. PMID- 14526044 TI - Infectious diseases. SARS experts want labs to improve safety practices. PMID- 14526045 TI - Astronomy. Smashup sends alien stars streaming near the sun. PMID- 14526047 TI - Women in science. Princeton study strikes sad but familiar chord. PMID- 14526046 TI - Ecology. 'Tragedy of the commons' author dies. PMID- 14526048 TI - Space exploration. Europe embarks on leisurely lunar odyssey. PMID- 14526049 TI - Endagered Species Act. Report says listings draw on sound science. PMID- 14526050 TI - 10 years after the SSC. Physics tries to leave the tunnel. PMID- 14526052 TI - 10 years after the SSC. Scientists are long gone, but bitter memories remain. PMID- 14526051 TI - 10 years after the SSC. Lots of reasons, but few lessons. PMID- 14526053 TI - 10 years after the SSC. The power of words: the SSC in literature. PMID- 14526054 TI - Chinese universities. An end to business as usual? PMID- 14526055 TI - Buddhism and neuroscience. Studying the well-trained mind. PMID- 14526056 TI - American Chemical Society meeting. Molecular scaffolding helps raise a crop of neurons. PMID- 14526057 TI - American Chemical Society meeting. Arraymaker speeds analyses by months. PMID- 14526058 TI - SARS, wildlife, and human health. PMID- 14526059 TI - How would a Cable Science Network work? PMID- 14526060 TI - A suggestion for the multiple author issue. PMID- 14526061 TI - A heterozygote advantage. PMID- 14526063 TI - Comment on "rapid evolution of egg size in captive salmon". PMID- 14526064 TI - Comment on "rapid evolution of egg size in captive Salmon". PMID- 14526066 TI - Medicine. The NIH Roadmap. AB - The NIH Roadmap is a set of bold initiatives aimed at accelerating medical research. These initiatives will address challenges that no single NIH institute could tackle alone, but the agency as a whole must undertake. The Roadmap identifies the most compelling opportunities in three arenas: new pathways to discovery, research teams of the future, and reengineering the clinical research enterprise. PMID- 14526068 TI - Physics. Driving the electron over the edge. AB - At low temperatures, many materials, from organics to oxides, undergo a pressure driven "Mott transition": A small change in the distance between the atoms causes the electrical resistivity of the system to change by many orders of magnitude, without a change in crystal symmetry. In his Perspective, Kotliar highlights the report by Limelette et al., who have investigated the dependence of the resistivity of a doped vanadium oxide on pressure and temperature around the Mott critical endpoint (where the Mott transition becomes continuous). A purely electronic model accounts for the major features of the experiments, but some more detailed features require the coupling between electrons and the crystal lattice to be taken into account. PMID- 14526069 TI - Chemistry. How to assemble a molecular junction. AB - Junctions between metals and molecules play an important role in molecular electronics. Advances in this field are hampered by the lack of understanding of the electronic structure of organic-metal interfaces. In his Perspective, Kummel highlights the report by Nazin et al. (3), who have used scanning tunneling microscopy (STM) to assemble a metal-molecule-metal junction. Subsequently, they employed the STM tip to probe the atomic structure and local electronic properties of the metal-molecule interface in unprecedented detail. They find evidence for strong coupling between the molecular and metal states. Such coupling affects the conductivity of metal-molecule-metal junctions. PMID- 14526067 TI - Neuroscience. Stout guards of the central nervous system. AB - Endocannabinoids have paradoxical effects on the mammalian nervous system: Sometimes they block neuronal excitability and other times they augment it. In their Perspective, Mechoulam and Lichtman discuss new work (Marsicano et al.) showing that activation of the cannabinoid receptor CB1 by the endocannabinoid anandamide protects against excitotoxic damage in a mouse model of kainic acid induced epilepsy. PMID- 14526070 TI - Chemistry. Toward vibrational mode control in catalysis. AB - Vibrational excitations of specific bonds in molecules have been used to enhance the reactivity of the molecules in direct gas-phase reactions. In his Perspective, Luntz highlights a report by Beck et al., who show that such vibrational control may also be possible for catalytic reactions at a surface. The authors demonstrate that differently excited deuterated methane molecules have different dissociation probabilities on a nickel surface, even though the energies of the different molecules are similar. PMID- 14526071 TI - Plant Biology. Hormones and the green revolution. AB - The success of the green revolution largely resulted from the creation of dwarf cultivars of wheat and rice, which had much higher yields than conventional crops. Characterization of these dwarf cultivars showed that the mutant genes were involved in either the synthesis or signaling of gibberellin, a plant growth hormone. In his Perspective, Salamini highlights new work (Multani et al.) that identifies the cause of dwarfism in agronomically important varieties of maize and sorghum. In these cases, dwarfism is caused by defective transport of another growth hormone called auxin. PMID- 14526072 TI - Genes record a prehistoric volcano eruption in the Galapagos. PMID- 14526073 TI - Loss of an MDR transporter in compact stalks of maize br2 and sorghum dw3 mutants. AB - Agriculturally advantageous reduction in plant height is usually achieved by blocking the action or production of gibberellins. Here, we describe a different dwarfing mechanism found in maize brachytic2 (br2) mutants characterized by compact lower stalk internodes. The height reduction in these plants results from the loss of a P-glycoprotein that modulates polar auxin transport in the maize stalk. The sorghum ortholog of br2 is dwarf3 (dw3), an unstable mutant of long standing commercial interest and concern. A direct duplication within the dw3 gene is responsible for its mutant nature and also for its instability, because it facilitates unequal crossing-over at the locus. PMID- 14526075 TI - Universality and critical behavior at the Mott transition. AB - We report conductivity measurements of Cr-doped V2O3 using a variable pressure technique. The critical behavior of the conductivity near the Mott insulator to metal critical endpoint is investigated in detail as a function of pressure and temperature. The critical exponents are determined, as well as the scaling function associated with the equation of state. The universal properties of a liquid-gas transition are found. This is potentially a generic description of the Mott critical endpoint in correlated electron materials. PMID- 14526074 TI - CB1 cannabinoid receptors and on-demand defense against excitotoxicity. AB - Abnormally high spiking activity can damage neurons. Signaling systems to protect neurons from the consequences of abnormal discharge activity have been postulated. We generated conditional mutant mice that lack expression of the cannabinoid receptor type 1 in principal forebrain neurons but not in adjacent inhibitory interneurons. In mutant mice,the excitotoxin kainic acid (KA) induced excessive seizures in vivo. The threshold to KA-induced neuronal excitation in vitro was severely reduced in hippocampal pyramidal neurons of mutants. KA administration rapidly raised hippocampal levels of anandamide and induced protective mechanisms in wild-type principal hippocampal neurons. These protective mechanisms could not be triggered in mutant mice. The endogenous cannabinoid system thus provides on-demand protection against acute excitotoxicity in central nervous system neurons. PMID- 14526076 TI - The anomalous Hall effect and magnetic monopoles in momentum space. AB - Efforts to find the magnetic monopole in real space have been made in cosmic rays and in particle accelerators, but there has not yet been any firm evidence for its existence because of its very heavy mass, approximately 10(16) giga-electron volts. We show that the magnetic monopole can appear in the crystal momentum space of solids in the accessible low-energy region (approximately 0.1 to 1 electron volts) in the context of the anomalous Hall effect. We report experimental results together with first-principles calculations on the ferromagnetic crystal SrRuO3 that provide evidence for the magnetic monopole in the crystal momentum space. PMID- 14526077 TI - Coherent soft x-ray generation in the water window with quasi-phase matching. AB - We demonstrate enhanced generation of coherent light in the "water window" region of the soft x-ray spectrum at 4.4 nanometers, using quasi-phase-matched frequency conversion of ultrafast laser pulses. By periodically modulating the diameter of a gas-filled hollow waveguide, the phase mismatch normally present between the laser light and the generated soft x-ray light can be partially compensated. This makes it possible to use neon gas as the nonlinear medium to coherently convert light up to the water window, illustrating that techniques of nonlinear optics can be applied effectively in the soft x-ray region of the spectrum. These results advance the prospects for compact coherent soft x-ray sources for applications in biomicroscopy and in chemical spectroscopy. PMID- 14526078 TI - Vibrational mode-specific reaction of methane on a nickel surface. AB - The dissociation of methane on a nickel catalyst is a key step in steam reforming of natural gas for hydrogen production. Despite substantial effort in both experiment and theory, there is still no atomic-scale description of this important gas-surface reaction. We report quantum state-resolved studies, using pulsed laser and molecular beam techniques, of vibrationally excited methane reacting on the nickel (100) surface. For doubly deuterated methane (CD2H2), we observed that the reaction probability with two quanta of excitation in one C-H bond was greater (by as much as a factor of 5) than with one quantum in each of two C-H bonds. These results clearly exclude the possibility of statistical models correctly describing the mechanism of this process and attest to the importance of full-dimensional calculations of the reaction dynamics. PMID- 14526079 TI - Identification of a plant nitric oxide synthase gene involved in hormonal signaling. AB - Nitric oxide (NO) serves as a signal in plants. An Arabidopsis mutant (Atnos1) was identified that had impaired NO production, organ growth, and abscisic acid induced stomatal movements. Expression of AtNOS1 with a viral promoter in Atnos1 mutant plants resulted in overproduction of NO. Purified AtNOS1 protein used the substrates arginine and nicotinamide adenine dinucleotide phosphate and was activated by Ca2+ and calmodulin-like mammalian endothelial nitric oxide synthase and neuronal nitric oxide synthase, yet it is a distinct enzyme with no sequence similarities to any mammalian isoform. Thus, AtNOS1 encodes a distinct nitric oxide synthase that regulates growth and hormonal signaling in plants. PMID- 14526080 TI - Talin binding to integrin beta tails: a final common step in integrin activation. AB - Control of integrin affinity for ligands (integrin activation) is essential for normal cell adhesion, migration, and assembly of an extracellular matrix. Integrin activation is usually mediated through the integrin beta subunit cytoplasmic tail and can be regulated by many different biochemical signaling pathways. We report that specific binding of the cytoskeletal protein talin to integrin beta subunit cytoplasmic tails leads to the conformational rearrangements of integrin extracellular domains that increase their affinity. Thus, regulated binding of talin to integrin beta tails is a final common element of cellular signaling cascades that control integrin activation. PMID- 14526081 TI - Self-assembly of proteins into designed networks. AB - A C4-symmetric tetrameric aldolase was used to produce a quadratic network consisting of the enzyme as a rigid four-way connector and stiff streptavidin rods as spacers. Each aldolase subunit was furnished with a His6 tag for oriented binding to a planar surface and two tethered biotins for binding streptavidin in an oriented manner. The networks were improved by starting with composite units and also by binding to nickel-nitrilotriacetic acid-lipid monolayers. The mesh was adjustable in 5-nanometer increments. The production of a net with switchable mesh was initiated with the use of a calcium ion-containing beta-helix spacer that denatured on calcium ion depletion. PMID- 14526082 TI - Untangling desmosomal knots with electron tomography. AB - Cell adhesion by adherens junctions and desmosomes relies on interactions between cadherin molecules. However, the molecular interfaces that define molecular specificity and that mediate adhesion remain controversial. We used electron tomography of plastic sections from neonatal mouse skin to visualize the organization of desmosomes in situ. The resulting three-dimensional maps reveal individual cadherin molecules forming discrete groups and interacting through their tips. Fitting of an x-ray crystal structure for C-cadherin to these maps is consistent with a flexible intermolecular interface mediated by an exchange of amino-terminal tryptophans. This flexibility suggests a novel mechanism for generating both cis and trans interactions and for propagating these adhesive interactions along the junction. PMID- 14526083 TI - Wild-type nonneuronal cells extend survival of SOD1 mutant motor neurons in ALS mice. AB - The most common inherited [correct] form of amyotrophic lateral sclerosis (ALS), a neurodegenerative disease affecting adult motor neurons, is caused by dominant mutations in the ubiquitously expressed Cu-Zn superoxide dismutase (SOD1). In chimeric mice that are mixtures of normal and SOD1 mutant-expressing cells, toxicity to motor neurons is shown to require damage from mutant SOD1 acting within nonneuronal cells. Normal motor neurons in SOD1 mutant chimeras develop aspects of ALS pathology. Most important, nonneuronal cells that do not express mutant SOD1 delay degeneration and significantly extend survival of mutant expressing motor neurons. PMID- 14526084 TI - Thalamic control of visceral nociception mediated by T-type Ca2+ channels. AB - Sensations from viscera, like fullness, easily become painful if the stimulus persists. Mice lacking alpha1G T-type Ca2+ channels show hyperalgesia to visceral pain. Thalamic infusion of a T-type blocker induced similar hyperalgesia in wild type mice. In response to visceral pain, the ventroposterolateral thalamic neurons evokeda surge of single spikes, which then slowly decayed as T type dependent burst spikes gradually increased. In alpha1G-deficient neurons, the single-spike response persisted without burst spikes. These results indicate that T-type Ca2+ channels underlie an antinociceptive mechanism operating in the thalamus andsupport the idea that burst firing plays a critical role in sensory gating in the thalamus. PMID- 14526085 TI - Performance monitoring by the anterior cingulate cortex during saccade countermanding. AB - Consensus is emerging that the medial frontal lobe of the brain is involved in monitoring performance, but precisely what is monitored remains unclear. A saccade-countermanding task affords an experimental dissociation of neural signals of error, reinforcement, and conflict. Single-unit activity was monitored in the anterior cingulate cortex of monkeys performing this task. Neurons that signaled errors were found, half of which responded to the omission of earned reinforcement. A further diversity of neurons signaled earned or unexpected reinforcement. No neurons signaled the form of conflict engendered by interruption of saccade preparation produced in this task. These results are consistent with the hypothesis that the anterior cingulate cortex monitors the consequences of actions. PMID- 14526086 TI - Evolutionary discrimination of mammalian conserved non-genic sequences (CNGs). AB - Analysis of the human and mouse genomes identified an abundance of conserved non genic sequences (CNGs). The significance and evolutionary depth of their conservation remain unanswered. We have quantified levels and patterns of conservation of 191 CNGs of human chromosome 21 in 14 mammalian species. We found that CNGs are significantly more conserved than protein-coding genes and noncoding RNAS (ncRNAs) within the mammalian class from primates to monotremes to marsupials. The pattern of substitutions in CNGs differed from that seen in protein-coding and ncRNA genes and resembled that of protein-binding regions. About 0.3% to 1% of the human genome corresponds to a previously unknown class of extremely constrained CNGs shared among mammals. PMID- 14526087 TI - Radar evidence for liquid surfaces on Titan. AB - Arecibo radar observations of Titan at 13-centimeter wavelength indicate that most of the echo power is in a diffusely scattered component but that a small specular component is present for about 75% of the subearth locations observed. These specular echoes have properties consistent with those expected for areas of liquid hydrocarbons. Knowledge of the areal extent and depth of any deposits of liquid hydrocarbons could strongly constrain the history of Titan's atmosphere and surface. PMID- 14526088 TI - Structure of the cytochrome b6f complex of oxygenic photosynthesis: tuning the cavity. AB - The cytochrome b6f complex provides the electronic connection between the photosystem I and photosystem II reaction centers of oxygenic photosynthesis and generates a transmembrane electrochemical proton gradient for adenosine triphosphate synthesis. A 3.0 angstrom crystal structure of the dimeric b6f complex from the thermophilic cyanobacterium Mastigocladus laminosus reveals a large quinone exchange cavity, stabilized by lipid, in which plastoquinone, a quinone-analog inhibitor, and a novel heme are bound. The core of the b6f complex is similar to the analogous respiratory cytochrome bc1 complex, but the domain arrangement outside the core and the complement of prosthetic groups are strikingly different. The motion of the Rieske iron-sulfur protein extrinsic domain, essential for electron transfer, must also be different in the b6f complex. PMID- 14526089 TI - Planetary science. The glitter of distant seas. AB - It has long been known that Saturn's largest moon, Titan, has a thick nitrogen atmosphere, which obscures the underlying surface. In his Perspective, Lorenz highlights the report by Campbell et al., who have used the giant Arecibo and Green Bank radio telescopes as a radar to probe Titan's hidden surface. The surface appears to be distinct from those of the icy satellites of Jupiter, in both brightness and polarization. The new data show sharp spikes in the reflected microwave spectrum, indicating large, smooth areas of radar-dark material. These features suggest the widespread existence of lakes or seas of liquid hydrocarbons on Titan. PMID- 14526090 TI - Quantification of bone involvement in Gaucher disease: MR imaging bone marrow burden score as an alternative to Dixon quantitative chemical shift MR imaging- initial experience. AB - PURPOSE: To develop a semiquantitative magnetic resonance (MR) imaging bone marrow burden (BMB) score with inclusion of both axial and peripheral bone marrow in Gaucher disease as an alternative to MR imaging with the Dixon quantitative chemical shift imaging (QCSI) technique. MATERIALS AND METHODS: Two experienced musculoskeletal radiologists with no experience in evaluating Gaucher disease blindly analyzed MR images of lumbar spines and femora. Interobserver and intraobserver variability were tested. In addition, the BMB score was determined as a parameter to evaluate bone marrow response to enzyme supplementation therapy. Finally, the BMB score was compared with fat fraction measurements obtained with Dixon QCSI. Differences between groups were analyzed by using the nonparametric Mann-Whitney test. A P value of less than.05 was considered to represent significance. Correlation was calculated by using two-tailed nonparametric rank correlation (Spearman rho). RESULTS: In 30 patients (mean age, 39.3 years; age range, 12-71 years) the mean fat fraction was 0.20 (range, 0.08 0.40). The BMB score range was 3-13 points. A significant correlation was found between the two observers when using BMB (rho = 0.91, P <.001). The intraobserver variation showed a significant correlation (rho = 0.99, P <.001). There was a significant correlation between BMB and QCSI (rho = -0.78, P <.001). Although BMB was less sensitive than Dixon QCSI, it showed enough sensitivity to allow detection of bone marrow response to enzyme supplementation therapy. CONCLUSION: BMB is a reproducible semiquantitative scoring system that is easy to use. It combines MR imaging of both axial and peripheral bone marrow and shows a significant correlation with QCSI. PMID- 14526091 TI - Contrast-enhanced US of microcirculation of superficially implanted tumors in rats. AB - PURPOSE: To determine the ability of contrast material-enhanced ultrasonography (US) to assess replenishment time in a rat kidney and adenocarcinoma tumor model. MATERIALS AND METHODS: Mammary adenocarcinoma cells were implanted into the subcutaneous tissues of the flank of 11 rats. Resultant tumors were imaged serially with contrast-enhanced US and compared with images of the rat kidney, a highly perfused normal organ. The US acquisition and processing methods yield images of perfused tumor regions and the times required to achieve 80% replenishment. Findings at contrast-enhanced computed tomography (CT) and light microscopy of hematoxylin-eosin-stained tumor tissue were compared. Paired Student t test was performed to compare the accuracy of US with that of histologic examination and CT in the detection of viable tumor regions. RESULTS: Replenishment of the kidney cortex microvasculature requires 1-5 seconds compared with a replenishment time of 6-14 seconds in tumors. Over the time course of tumor growth, the mean perfusion time becomes progressively longer, and a wider range of perfusion times is detected. Comparison of findings at US, CT, and histologic examination suggested that all three methods yield correlated estimates of the percentage of viable perfused tumor cells. Results of the t test suggested that the viable tumor percentages observed at US are not significantly different from those observed at CT and histologic examination (US vs CT, P =.92; US vs histologic examination, P =.94). CONCLUSION: Repeated measurements of microvascular flow rate can be accomplished in a rat animal model with a minimally invasive technique. PMID- 14526092 TI - Real-time MR imaging of aortic flow: influence of breathing on left ventricular stroke volume in chronic obstructive pulmonary disease. AB - PURPOSE: To assess real-time changes of left ventricular stroke volume (SV) in relation to the breathing pattern in healthy subjects and in patients with chronic obstructive pulmonary disease (COPD). MATERIALS AND METHODS: Real-time magnetic resonance (MR) imaging flow measurements were performed in the ascending aorta of 10 healthy volunteers and nine patients with severe COPD. Breathing maneuvers were registered with an abdominal pressure belt, which was synchronized to the electrocardiographic signal and the flow measurement. Healthy subjects performed normal breathing, deep breathing, and the Valsalva maneuver. Patients with COPD performed spontaneous breathing. Paired two-tailed Student t tests were used in healthy volunteers to assess significant SV differences between normal breathing and deep breathing or the Valsalva maneuver. The results of measurements in the patients with COPD were compared with the results during normal breathing in healthy subjects with the unpaired two-tailed Student t test. RESULTS: In healthy subjects, SV decreased during inspiration and increased during expiration (r2 = 0.78, P <.05). When compared with the SV during normal breathing, mean SV did not change during deep breathing but declined during the Valsalva maneuver (P <.05). The difference between minimal and maximal SVs (ie, SV range) increased because of deep breathing or the Valsalva maneuver (P <.05). In normal and deep breathing, velocity of SV elevation and velocity of SV decrease were equal (which resulted in a ratio of 1), whereas during the Valsalva maneuver, this value increased (P <.05). Spontaneous breathing in COPD resulted in SV changes (P <.05) similar to those observed in healthy subjects who performed the Valsalva maneuver. CONCLUSION: Real-time MR imaging of aortic flow reveals physiologic flow alterations, which are dependent on variations in the breathing pattern. PMID- 14526093 TI - Vascular calcification in ex vivo carotid specimens: precision and accuracy of measurements with multi-detector row CT. AB - PURPOSE: To test the accuracy and precision of multi-detector row computed tomography (CT)-derived measurements of vascular calcification in ex vivo human carotid endarterectomy (CEA) specimens. MATERIALS AND METHODS: Sixteen ex vivo CEA specimens were imaged with multi-detector row CT. Multi-detector row CT derived calcium scoring algorithms (ie, mineral mass and volume score) were compared with the mass and volume of ashed remnants of the CEA specimens. Bland Altman analysis was performed to assess the mean (ie, bias) and SD (ie, precision) of differences between multi-detector row CT- and ashing-derived measurements. In addition, conventional Agatston score, volume score, mineral mass, and modified Agatston score were calculated for each specimen by using a number of scanning protocols. Images were obtained at a section thickness of 1.25 mm by using different tube energy settings and tube currents. Specimens were also imaged at different section thicknesses with fixed combinations of tube energy and tube current. To compare measurement variability of scoring methods, coefficients of variation for all protocols were calculated. RESULTS: Both mean multi-detector row CT-derived mineral mass and mean ashing-derived mineral mass were 0.129 g +/- 0.173 (r = 0.99, P <.001). Mean multi-detector row CT- and ashing-derived volumes were 339.94 mm3 +/- 395.4 and 39.48 mm3 +/- 55.76, respectively (r = 0.95, P <.001). Measurement bias relative to the reference ashing values was high (2,800.0%) for volume score and low (2.58%) for mineral mass. Measurement precision was 0.6% for volume score and greater than 0.0005% for mineral mass. Mean coefficients of variation for all CT protocols were 5.0% +/- 4.2 and 4.9% +/- 4.2 for mineral mass and modified Agatston score, respectively, and 16.0% +/- 9.2 and 14.5% +/- 3.9 for conventional Agatston and volume scores, respectively (P <.001). CONCLUSION: Compared with the conventional volume score, multi-detector row CT-derived mineral mass is a less biased and more precise measurement of the mineral content of nonmoving ex vivo CEA specimens. Mineral mass and modified Agatston score are more reproducible than conventional volume and Agatston scores. PMID- 14526094 TI - Human gallbladder bile: noninvasive investigation in vivo with single-voxel 1H MR spectroscopy. AB - Proton (hydrogen 1) magnetic resonance (MR) spectroscopy was used to study model and porcine bile in vitro. The method was subsequently developed to facilitate the acquisition of in vivo 1H MR spectra from the gallbladder bile of 10 human volunteers. Signals attributable to phosphotidylcholine and conjugated bile acid protons were observed in eight of the 10 volunteers. Phosphotidylcholine concentrations were estimated, and five values (mean = 35.8 mmol/L, SD = 9.8) were within the expected range of levels in human bile. Findings in this preliminary investigation indicate that human gallbladder bile can be qualitatively and quantitatively studied noninvasively with 1H MR spectroscopy. PMID- 14526095 TI - Helical CT for nephrolithiasis and ureterolithiasis: comparison of conventional and reduced radiation-dose techniques. AB - PURPOSE: To determine the accuracy of unenhanced helical computed tomography (CT) performed at reduced milliampere-second, and therefore at a reduced patient radiation dose, by using conventional unenhanced helical CT as the standard. MATERIALS AND METHODS: Fifty patients with acute flank pain who weighed less than 200 lb (90 kg) were prospectively recruited for this study. Conventional helical CT scans were obtained with patients in the prone position by using 5-mm-thick sections, 140 kVp, 135-208 mAs (mean, 160 mAs), and a pitch of 1.5 (single detector row CT) or 0.75 (multi-detector row CT, 4 x 5-mm detector configuration). Conventional CT was immediately followed by low-dose scanning, whereby the tube current was reduced to 100 mA (mean, 76 mAs). All other technical parameters and anatomic coverage remained constant. Three independent readers who were blinded to patient identity interpreted the scans in random order. The observers noted the location, size, and number of calculi; secondary signs of obstruction; and other clinically relevant findings. High- and low-dose scans were compared by using paired t tests and the signed rank test. RESULTS: Calculi were found in 33 (66%) patients; 25 (50%) had renal calculi and 19 (38%) had an obstructing ureteral calculus. The accuracy rates (averaged over the three readers) for determining the various findings on the low-dose scan compared with the high-dose scan were as follows: nephrolithiasis, 91%; ureterolithiasis, 94%; obstruction, 91%; and normal findings, 92%. When interpretations between readers were compared, agreement rates were 90%-95% for standard-dose scans and 90%-92% for reduced-dose scans (P >.5). Uncomplicated mild diverticulitis was found in three patients. No other clinically important abnormality was identified. A reduction in the tube current to 100 mA resulted in a dose reduction of 25% for multi-detector row CT and 42% for single-detector row CT. CONCLUSION: In patients who weighed less than 200 lb, unenhanced helical CT performed at a reduced tube current of 100 mA, and therefore at a reduced patient dose, resulted in scans of high accuracy. PMID- 14526096 TI - Hypertrophic pyloric stenosis in infants: US evaluation of vascularity of the pyloric canal. AB - PURPOSE: To determine if there is increased flow to the pylorus in infants with hypertrophic pyloric stenosis (HPS) and, if so, whether the flow is localized to the muscle layer, mucosal layer, or both. MATERIALS AND METHODS: Seventy-five infants examined for clinical suspicion of HPS were prospectively recruited for the study. Color scale was standardized at 4.2-4.4 cm/sec. Color Doppler flow at ultrasonography (US) was graded as follows: Grade 1 meant no signal; grade 2, two to five flow signals; and grade 3, extensive or continuous flow. Flow to the muscle or mucosal layer was documented and confirmed with spectral analysis. Infants without HPS served as control patients. Descriptive analyses were conducted to assess the demographic data and US results. Significance was assessed with chi2 or t tests. P <.05 was considered to indicate a significant difference. RESULTS: HPS was present in 41 infants with a mean age of 5 weeks +/- 2.0 (SD). Their mean flow grade was 2.80 +/- 0.4 in muscle and 2.88 +/- 0.4 in mucosa. HPS was not present in 34 infants with a mean age of 5.9 weeks +/- 4.5. Their mean flow grade was 1.26 +/- 0.5 in muscle and 1.15 +/- 0.5 in mucosa (P <.001). There was no significant difference in flow grades when the dimensions of the pyloric muscle and mucosa were compared. There was no significant difference in age between the HPS and control patient groups. CONCLUSION: Increased flow accompanies and may conceivably represent an integral component of the changes that occur with infantile HPS. PMID- 14526097 TI - Three-dimensional volume-rendered helical CT before laparoscopic adrenalectomy. AB - Use of three-dimensional (3D) volume-rendered helical computed tomography (CT) in surgical planning before laparoscopic adrenalectomy was evaluated in a retrospective study. In 35 consecutive patients before laparoscopic adrenalectomy, 3D volume-rendered CT scans were created from helical CT scans. Videotapes that showed anterior, lateral, posterior, and posterocephalic approaches were assessed retrospectively. The relationship (not contacting, abutting, displacing, or invading) of adrenal masses to adjacent organs (diaphragm, liver, spleen, kidneys, stomach, pancreas, and vessels) was recorded and compared with findings in surgery reports. When such findings were available, they corresponded to those in the videotape. Three-dimensional volume-rendered CT successfully displayed the relationship of adrenal masses to adjacent anatomic structures and organs before laparoscopic adrenalectomy. PMID- 14526098 TI - Severe acute respiratory syndrome: relationship between radiologic and clinical parameters. AB - PURPOSE: To quantify severity of severe acute respiratory syndrome (SARS) on chest radiographs and to determine its relationship with clinical parameters. MATERIALS AND METHODS: Forty patients (mean age, 42.90 years +/- 14.01 [SD]; median age, 41.5 years; age range, 25-82 years) with clinically diagnosed SARS were evaluated. Heart rate, oxygen saturation, temperature, and alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were recorded daily. Severity of lung changes on chest radiographs was scored according to percentage of involved lung. Radiographic scores at days of admission, treatment, and maximal radiographic score were extracted for statistical analysis with clinical parameters. Time to maximal radiographic score from admission and days between onset and beginning of treatment were determined. Correlations between radiographic and clinical parameters were evaluated with Spearman rank correlation. Sex differences with respect to clinical and radiographic parameters were evaluated with Mann-Whitney test. RESULTS: Median chest radiographic scores peaked 5 days after beginning of treatment before they declined. Maximal and treatment radiographic scores were inversely related to oxygen saturation (r = 0.67, P <.001; r = -0.35, P =.03). Admission radiographic score was correlated with admission AST level (r = 0.53, P =.003); treatment radiographic score, with treatment ALT and AST levels (r = 0.43, P =.007; r = 0.42, P =.019); and time to maximal radiographic score, with AST level at maximal radiographic score (r = 0.45, P =.006), admission radiographic score (r = -0.55, P <.001), treatment radiographic score (r = -0.58, P <.001), and admission ALT and AST levels (r = 0.44, P =.007; r = -0.58, P =.001). Treatment delay was associated with AST level at maximal radiographic score (r = 0.53, P =.001), treatment radiographic score (r = 0.60, P <.001), and time to maximal radiographic score (r = -0.36, P =.02). No sex differences occurred with respect to radiographic and clinical parameters (P >.05). CONCLUSION: Severity of lung abnormalities quantified on chest radiographs correlates with clinical and laboratory parameters. PMID- 14526099 TI - Matching protein structures with fuzzy alignments. AB - Unraveling functional and ancestral relationships between proteins as well as structure-prediction procedures require powerful protein-alignment methods. A structure-alignment method is presented where the problem is mapped onto a cost function containing both fuzzy (Potts) assignment variables and atomic coordinates. The cost function is minimized by using an iterative scheme, where at each step mean field theory methods at finite "temperatures" are used for determining fuzzy assignment variables followed by exact translation and rotation of atomic coordinates weighted by their corresponding fuzzy assignment variables. The approach performs very well when compared with other methods, requires modest central processing unit consumption, and is robust with respect to choice of iteration parameters for a wide range of proteins. PMID- 14526100 TI - A multitude of genes expressed solely in meiotic or postmeiotic spermatogenic cells offers a myriad of contraceptive targets. AB - Understanding mammalian spermatozoan development and the events surrounding fertilization has grown slowly, in part because of uncertainty about the number and identity of the cellular components involved. Determination of those transcripts expressed specifically by germ cells should provide an inclusive list of probable critical proteins. Here, total mouse testis transcript profiles were trimmed of transcripts found in cultures enriched in Sertoli or interstitial cells to yield a germ cell-enriched transcript profile. Monitoring of changes of this profile in the developing testis identified 1,652 genes whose transcript abundance increased markedly coincident with the onset of meiosis. Remarkably, 351 of these genes (approximately equal to 20%) appear to be expressed only in the male germline. Germ cell-specific transcripts are much less common earlier in testis development. Further analysis of the UniGene EST database coupled with quantitative PCR indicates that approximately 4% of the mouse genome is dedicated to expression in postmeiotic male germ cells. Most or many of the protein products of these transcripts are probably retained in mature spermatozoa. Targeted disruption of 19 of these genes has indicated that a majority have roles critical for normal fertility. Thus, we find an astonishing number of genes expressed specifically by male germ cells late in development. This extensive group provides a plethora of potential targets for germ cell-directed contraception and a staggering number of candidate proteins that could be critical for fertilization. PMID- 14526101 TI - Sequential megafaunal collapse in the North Pacific Ocean: an ongoing legacy of industrial whaling? AB - Populations of seals, sea lions, and sea otters have sequentially collapsed over large areas of the northern North Pacific Ocean and southern Bering Sea during the last several decades. A bottom-up nutritional limitation mechanism induced by physical oceanographic change or competition with fisheries was long thought to be largely responsible for these declines. The current weight of evidence is more consistent with top-down forcing. Increased predation by killer whales probably drove the sea otter collapse and may have been responsible for the earlier pinniped declines as well. We propose that decimation of the great whales by post World War II industrial whaling caused the great whales' foremost natural predators, killer whales, to begin feeding more intensively on the smaller marine mammals, thus "fishing-down" this element of the marine food web. The timing of these events, information on the abundance, diet, and foraging behavior of both predators and prey, and feasibility analyses based on demographic and energetic modeling are all consistent with this hypothesis. PMID- 14526102 TI - Angiogenesis impairment in Id-deficient mice cooperates with an Hsp90 inhibitor to completely suppress HER2/neu-dependent breast tumors. AB - Id proteins bind basic helix-loop-helix transcription factors and function as dominant negative inhibitors of gene expression. Id1 and Id3 are required for the recruitment of bone marrow-derived endothelial cell precursors and tumors transplanted into Id-deficient mice demonstrate impaired angiogenesis. Mouse mammary tumor virus-neu mice were bred with Id1-/-Id3+/- mice to ascertain the role of Id1 and Id3 in mammary tumorigenesis in a more physiologically relevant model. In mammary tumors from these mice, Id1 and Id3 expression was restricted to the vascular endothelium. Id1 and Id3 deficiency did not prevent or delay tumor formation but did alter tumor phenotype. The tumors that developed in the Id-deficient mice were larger and cystic with a viable rim of tumor cells surrounding a nonviable core of cellular debris. The Hsp90 chaperone protein is required for cellular survival under condition of environmental stress and for the stability of the neu oncogene. 17-Allylamino-17-demethoxygeldanamycin, an Hsp90 inhibitor, was used to treat these mice. Whereas 17-allylamino-17 demethoxygeldanamycin only modestly delayed the growth of established mammary tumors in WT mice for Id, tumor suppression was dramatically more effective in an Id1- or Id3-deficient background. These data suggest that tumorigenesis can occur in a background of defective angiogenesis but that tumors developing in such an environment may be especially sensitive to inhibitors of neu and stress-activated survival pathways. Thus angiogenesis inhibitors in combination with inhibitors of Hsp90 function should be evaluated for the treatment of advanced breast cancer. PMID- 14526103 TI - Invasion-inhibitory antibodies inhibit proteolytic processing of apical membrane antigen 1 of Plasmodium falciparum merozoites. AB - Apical membrane antigen 1 (AMA-1) is a promising vaccine candidate for Plasmodium falciparum malaria. Antibodies against AMA-1 of P. falciparum (PfAMA-1) interrupt merozoite invasion into RBCs. Initially localized within the apical complex, PfAMA-1 is proteolytically processed and redistributed circumferentially on merozoites at about the time of their release and invasion into RBCs. An 83-kDa precursor form of PfAMA-1 is processed to 66-kDa and then to 48- and 44-kDa products. We show that, even at low concentrations, IgG antibodies against correctly folded recombinant PfAMA-1 cross-linked and trapped the 52-, 48-, and 44-kDa proteolytic products on merozoites. These products are normally shed into the culture medium. At higher concentrations antibodies inhibited invasion into RBCs and caused a reduction in the amount of 44- and 48-kDa products, both on merozoites and in the culture medium. A corresponding increase also occurred in the amount of the 66- and 52-kDa forms detected on the merozoites. These antibodies also prevented circumferential redistribution of AMA-1. In contrast, monovalent invasion-inhibitory Fab fragments caused accumulation of 66- and 52 kDa forms, with no cross-linking, trapping, or prevention of redistribution. Antibodies at low concentrations can be used as trapping agents for intermediate and soluble forms of AMA-1 and are useful for studying proteolytic processing of AMA-1. With this technique, it was confirmed that protease inhibitor chymostatin and Ca2+ chelators can inhibit the breakdown of the 66-kDa form. We propose that antibodies to AMA-1 capable of inhibiting erythrocyte invasion act by disrupting proteolytic processing of AMA-1. PMID- 14526104 TI - Sp8 is crucial for limb outgrowth and neuropore closure. AB - In this report we describe the developmental expression and function of Sp8, a member of the Sp family of zinc finger transcription factors, and provide evidence that the legless transgene insertional mutant is a hypomorphic allele of the Sp8 gene. Sp8 is expressed during embryogenesis in the forming apical ectodermal ridge (AER), restricted regions of the central nervous system, and tail bud. Targeted deletion of the Sp8 gene gives a striking phenotype, with severe truncation of both forelimbs and hindlimbs, absent tail, as well as defects in anterior and posterior neuropore closure leading to exencephaly and spina bifida. Outgrowth of the limb depends on formation of the AER, a signaling center that forms at the limb bud apex. In Sp8 mutants, the AER precursor cells are induced and initially express multiple appropriate marker genes, but expression of these genes is not maintained and progression to a mature AER is blocked. These observations indicate that Sp8 functions downstream of Wnt3, Fgf10, and Bmpr1a in the signaling cascade that mediates AER formation. PMID- 14526105 TI - Brassinosteroids promote root growth in Arabidopsis. AB - Although brassinosteroids (BRs) are known to regulate shoot growth, their role in the regulation of root growth is less clear. We show that low concentrations of BRs such as 24-epicastasterone and 24-epibrassinolide promote root elongation in Arabidopsis wild-type plants up to 50% and in BR-deficient mutants such as dwf1-6 (cbb1) and cbb3 (which is allelic to cpd) up to 150%. The growth-stimulating effect of exogenous BRs is not reduced by the auxin transport inhibitor 2,3,5 triidobenzoic acid. BR-deficient mutants show normal gravitropism, and 2,3,5 triidobenzoic acid or higher concentrations of 2,4-dichlorophenoxyacetic acid and naphtaleneacetic acid inhibit root growth in the mutants to the same extent as in wild-type plants. Simultaneous administration of 24-epibrassinolide and 2,4 dichlorophenoxyacetic acid results in largely additive effects. Exogenous gibberellins do not promote root elongation in the BR-deficient mutants, and the sensitivity to the ethylene precursor 1-aminocyclopropane-1-carboxylic acid is not altered. Thus, the root growth-stimulating effect of BRs appears to be largely independent of auxin and gibberellin action. Furthermore, we analyzed BR interactions with other phytohormones on the gene expression level. Only a limited set of auxin- and ethylene-related genes showed altered expression levels. Genes related to other phytohormones barely showed changes, providing further evidence for an autonomous stimulatory effect of BR on root growth. PMID- 14526106 TI - Vascularization, high-volume solution flow, and localized roles for enzymes of sucrose metabolism during tumorigenesis by Agrobacterium tumefaciens. AB - Vascular differentiation and epidermal disruption are associated with establishment of tumors induced by Agrobacterium tumefaciens. Here, we address the relationship of these processes to the redirection of nutrient-bearing water flow and carbohydrate delivery for tumor growth within the castor bean (Ricinus communis) host. Treatment with aminoethoxyvinyl-glycine showed that vascular differentiation and epidermal disruption were central to ethylene-dependent tumor establishment. CO2 release paralleled tumor growth, but water flow increased dramatically during the first 3 weeks. However, tumor water loss contributed little to water flow to host shoots. Tumor water loss was followed by accumulation of the osmoprotectants, sucrose (Suc) and proline, in the tumor periphery, shifting hexose-to-Suc balance in favor of sugar signals for maturation and desiccation tolerance. Concurrent activities and sites of action for enzymes of Suc metabolism changed: Vacuolar invertase predominated during initial import of Suc into the symplastic continuum, corresponding to hexose concentrations in expanding tumors. Later, Suc synthase (SuSy) and cell wall invertase rose in the tumor periphery to modulate both Suc accumulation and descending turgor for import by metabolization. Sites of abscisic acid immunolocalization correlated with both central vacuolar invertase and peripheral cell wall invertase. Vascular roles were indicated by SuSy immunolocalization in xylem parenchyma for inorganic nutrient uptake and in phloem, where resolution allowed SuSy identification in sieve elements and companion cells, which has widespread implications for SuSy function in transport. Together, data indicate key roles for ethylene-dependent vascularization and cuticular disruption in the redirection of water flow and carbohydrate transport for successful tumor establishment. PMID- 14526107 TI - eEF1A isoforms change in abundance and actin-binding activity during maize endosperm development. AB - Eukaryotic elongation factor 1A (eEF1A) appears to be a multifunctional protein because several biochemical activities have been described for this protein, in addition to its role in protein synthesis. In maize (Zea mays) endosperm, the synthesis of eEF1A is increased in o2 (opaque2) mutants, and its concentration is highly correlated with the protein-bound lysine content. To understand the basis of this relationship, we purified eEF1A isoforms from developing endosperm and investigated their accumulation and their functional and structural properties. Formation of three isoforms appears to be developmentally regulated and independent of the o2 mutation, although one isoform predominated in one high lysine o2 inbred. The purified proteins differ in their ability to bind F-actin in vitro, suggesting that they are functionally distinct. However, they share similar aminoacyl-tRNA-binding activities. Tandem mass spectrometry revealed that each isoform is composed of the four same gene products, which are modified posttranslationally by methylation and phosphorylation. The chemical differences that account for their different actin-binding activities could not be determined. PMID- 14526108 TI - p-Chlorophenoxyisobutyric acid impairs auxin response in Arabidopsis root. AB - p-Chlorophenoxyisobutyric acid (PCIB) is known as a putative antiauxin and is widely used to inhibit auxin action, although the mechanism of PCIB-mediated inhibition of auxin action is not characterized very well at the molecular level. In the present work, we showed that PCIB inhibited BA::beta-glucuronidase (GUS) expression induced by indole-3-acetic acid (IAA), 2,4-dichlorophenoxyacetic acid, and 1-naphthaleneacetic acid. PCIB also inhibited auxin-dependent DR5::GUS expression. RNA hybridization and quantitative reverse transcriptase-polymerase chain reaction analyses suggested that PCIB reduced auxin-induced accumulation of transcripts of Aux/IAA genes. In addition, PCIB relieved the reduction of GUS activity in HS::AXR3NT-GUS transgenic line in which auxin inhibits GUS activity by promoting degradation of the AXR3NT-GUS fusion protein. Physiological analysis revealed that PCIB inhibited lateral root production, gravitropic response of roots, and growth of primary roots. These results suggest that PCIB impairs auxin signaling pathway by regulating Aux/IAA protein stability and thereby affects the auxin-regulated Arabidopsis root physiology. PMID- 14526109 TI - Plasma membrane aquaporins are involved in winter embolism recovery in walnut tree. AB - In perennial plants, freeze-thaw cycles during the winter months can induce the formation of air bubbles in xylem vessels, leading to changes in their hydraulic conductivity. Refilling of embolized xylem vessels requires an osmotic force that is created by the accumulation of soluble sugars in the vessels. Low water potential leads to water movement from the parenchyma cells into the xylem vessels. The water flux gives rise to a positive pressure essential for the recovery of xylem hydraulic conductivity. We investigated the possible role of plasma membrane aquaporins in winter embolism recovery in walnut (Juglans regia). First, we established that xylem parenchyma starch is converted to sucrose in the winter months. Then, from a xylem-derived cDNA library, we isolated two PIP2 aquaporin genes (JrPIP2,1 and JrPIP2,2) that encode nearly identical proteins. The water channel activity of the JrPIP2,1 protein was demonstrated by its expression in Xenopus laevis oocytes. The expression of the two PIP2 isoforms was investigated throughout the autumn-winter period. In the winter period, high levels of PIP2 mRNA and corresponding protein occurred simultaneously with the rise in sucrose. Furthermore, immunolocalization studies in the winter period show that PIP2 aquaporins were mainly localized in vessel-associated cells, which play a major role in controlling solute flux between parenchyma cells and xylem vessels. Taken together, our data suggest that PIP2 aquaporins could play a role in water transport between xylem parenchyma cells and embolized vessels. PMID- 14526110 TI - Functional and phylogenetic analyses of a conserved regulatory program in the phloem of minor veins. AB - The minor-vein phloem of mature leaves is developmentally and physiologically distinct from the phloem in the rest of the vascular system. Phloem loading of transport sugars occurs in the minor veins, and consistent with this, galactinol synthase is expressed in the minor veins of melon (Cucumis melo) as part of the symplastic-loading mechanism that operates in this species. A galactinol synthase promoter from melon drives gene expression in the minor-vein companion cells of both transgenic tobacco (Nicotiana tabacum) and Arabidopsis. Neither of these plants use galactinol in the phloem-loading process, implying that the promoter responds to a minor-vein-specific regulatory cascade that is highly conserved across a broad range of eudicotyledons. Detailed analysis of this promoter by truncation and mutagenesis identified three closely coupled sequences that unambiguously modulate tissue specificity. These sequences cooperate in a combinatorial fashion: two promote expression throughout the vascular system of the plant, whereas the third functions to repress expression in the larger bundles. In a complementary approach, phylogenetic footprinting was used to obtain single-nucleotide resolution of conserved sites in orthologous promoters from diverse members of the Cucurbitaceae. This comparative analysis confirmed the importance of the closely coupled sites but also revealed other highly conserved sequences that may modulate promoter strength or contribute to expression patterns outside of the phloem. The conservation of this regulatory design among species that phloem load by different mechanisms supports a model for organismal development in which tissues and cell types are controlled by relatively ancient and conserved paradigms but expression of genes influencing final form and function are relatively plastic. PMID- 14526111 TI - Resorption protection. Anthocyanins facilitate nutrient recovery in autumn by shielding leaves from potentially damaging light levels. AB - The resorption protection hypothesis, which states that anthocyanins protect foliar nutrient resorption during senescence by shielding photosynthetic tissues from excess light, was tested using wild-type (WT) and anthocyanin-deficient mutants of three deciduous woody species, Cornus sericea, Vaccinium elliottii (Chapmn.), and Viburnum sargentii (Koehne). WT Betula papyrifera (Marsh) was included to compare the senescence performance of a species that does not produce anthocyanins in autumn. Plants were subjected to three environmental regimes during senescence: an outdoor treatment; a 5-d high-stress (high light and low temperature) treatment followed by transfer to a low-stress environment and a low stress treatment that served as control. In the outdoor treatment, the appearance of anthocyanins in senescing leaves of WT plants was concomitant with the development of photo-inhibition in mutant plants of all three anthocyanin producing species. In the high-stress environment, WT plants maintained higher photochemical efficiencies than mutants and were able to recover when transferred to the low-stress environment, whereas mutant leaves dropped while still green and displayed signs of irreversible photooxidative damage. Nitrogen resorption efficiencies and proficiencies of all mutants in both stressful treatments were significantly lower than the WT counterparts. B. papyrifera displayed photochemical efficiencies and nitrogen resorption performance comparable with the highest of the anthocyanin-producing species in all three senescing environments, indicating a photoprotective strategy divergent from the other species studied. These results strongly support the resorption protection hypothesis of anthocyanins in senescing leaves. PMID- 14526113 TI - Interaction of cadmium with glutathione and photosynthesis in developing leaves and chloroplasts of Phragmites australis (Cav.) Trin. ex Steudel. AB - We investigated how the presence of cadmium (Cd) at the emergence of Phragmites australis Trin. (Cav.) ex Steudel plants from rhizomes interacted with leaf and chloroplast physiological and biochemical processes. About 8.5 nmol Cd mg-1 chlorophyll was found in leaves, and 0.83 nmol Cd mg-1 chlorophyll was found in chloroplasts of plants treated with 50 microm Cd. As a result, a 30% loss of chlorophyll was measured concomitantly with a comparable percentage reduction in light-saturated photosynthesis. Rubisco content and activity were lowered by 10% and 60%, respectively. Antioxidant activity was stimulated by Cd treatment and was associated with an increase in the glutathione and pyridine pools, and with a larger pool of reduced glutathione. It is suggested that the glutathione pool and its predominance in the reduced state protected the activity of many key photosynthetic enzymes against the thiophilic binding of Cd. Chloroplast ultrastructure was not significantly altered with 50 microm treatment and the efficiency of photosystem II, measured as the fluorescence ratio Fv/Fm, remained high because F0 and Fm were proportionally decreased. In plants treated with 100 microm Cd, all effects were exacerbated, but Fv/Fm remained close to that of control leaves and the glutathione and pyridine nucleotides pools were lowered. The results suggest that glutathione exerted a direct important protective role on photosynthesis in the presence of Cd. PMID- 14526112 TI - Two lily SEPALLATA-like genes cause different effects on floral formation and floral transition in Arabidopsis. AB - Two AGL2-like MADS-box genes, Lily MADS Box Gene (LMADS) 3 and LMADS4, with extensive homology of LMADS3 to the Arabidopsis SEPALLATA3 were characterized from the lily (Lilium longiflorum). Both LMADS3 and LMADS4 mRNA were detected in the inflorescence meristem, in floral buds of different developmental stages, and in all four whorls of the flower organ. LMADS4 mRNA is also expressed in vegetative leaf and in the inflorescence stem where LMADS3 expression is absent. Transgenic Arabidopsis, which ectopically expresses LMADS3, showed novel phenotypes by significantly reducing plant size, flowering extremely early, and loss of floral determinacy. By contrast, 35S::LMADS4 transgenic plants were morphologically indistinguishable from wild-type plants. The early-flowering phenotype in 35S::LMADS3 transgenic Arabidopsis plants was correlated with the up regulation of flowering time genes FT, SUPPRESSOR OF OVEREXPRESSION OF CO 1, LUMINIDEPENDENS, and flower meristem identity genes LEAFY and APETALA1. This result was further supported by the ability of 35S::LMADS3 to rescue the late flowering phenotype in gigantea-1 (gi-1), constans-3 (co-3), and luminidependens 1 but not for ft-1 or fwa-1 mutants. The activation of these flowering time genes is, however, indirect because their expression was unaffected in plants transformed with LMADS3 fused with rat glucocorticoid receptor in the presence of both dexamethasone and cycloheximide. PMID- 14526114 TI - Cleavage of the Pseudomonas syringae type III effector AvrRpt2 requires a host factor(s) common among eukaryotes and is important for AvrRpt2 localization in the host cell. AB - Many phytopathogenic bacteria use a type III secretion system to deliver type III effector proteins into the host plant cell. The Pseudomonas syringae type III effector AvrRpt2 is cleaved at a specific site when translocated into the host cell. In this study, we first demonstrate that the factor(s) required for AvrRpt2 cleavage is present in extracts from animal and yeast cells, as well as plant cells. The cleavage factor in animal and plant cell extracts was heat labile but relatively insensitive to protease inhibitors. Second, mutational analysis of AvrRpt2 was applied to identify features important for its cleavage. In addition to two of the amino acid residues in the immediate vicinity of the cleavage site, a large part of the region C-terminal to the cleavage site was required when AvrRpt2 was cleaved in animal cell extract. Most of these features were also important when AvrRpt2 was cleaved in plant cells. Third, we investigated the effect of cleavage in interactions of AvrRpt2 with plant cells. Cleavage of AvrRpt2 appeared to be important for proper interactions with Arabidopsis cells that lack the resistance gene product corresponding to AvrRpt2, RPS2. In addition, removal of the region N-terminal to the cleavage site was important for the correct localization of the C-terminal effector region of the protein in the host cell. We speculate that the virulence function of AvrRpt2 requires removal of the N-terminal region to redirect the effector protein to a specific subcellular location in the host cell after translocation of the protein. PMID- 14526115 TI - Characterization of mannuronan C-5-epimerase genes from the brown alga Laminaria digitata. AB - Alginate is an industrially important polysaccharide obtained commercially by harvesting brown algae. The final step in alginate biosynthesis, the epimerization of beta-1,4-d-mannuronic acid to alpha-1,4-l-guluronic acid, a structural change that controls the physicochemical properties of the alginate, is catalyzed by the enzyme mannuronan C-5-epimerase. Six different cDNAs with homology to bacterial mannuronan C-5-epimerases were isolated from the brown alga Laminaria digitata (Phaeophyceae). Hydrophobic cluster analysis indicated that the proteins encoded by the L. digitata sequences have important structural similarities to the bacterial mannuronan C-5-epimerases, including conservation of the catalytic site. The expression of the C-5-epimerase genes was examined by northern-blot analysis and reverse transcriptase-polymerase chain reaction in L. digitata throughout a year. Expression was also monitored in protoplast cultures by northern and western blot, reverse transcriptase-polymerase chain reaction, and activity measurements. From both the structural comparisons and the expression pattern, it appears that the cDNAs isolated from L. digitata encode functional mannuronan C-5-epimerases. The phylogenetic relationships of the bacterial and brown algal enzymes and the inferences on the origin of alginate biosynthetic machinery are discussed. PMID- 14526116 TI - Seed 1-cysteine peroxiredoxin antioxidants are not involved in dormancy, but contribute to inhibition of germination during stress. AB - Peroxiredoxins (Prx) are thiol-dependent antioxidants containing one (1-cysteine [-Cys]) or two (2-Cys) conserved Cys residues that protect lipids, enzymes, and DNA against reactive oxygen species. In plants, the 1-Cys Prxs are highly expressed during late seed development, and the expression pattern is dormancy related in mature seeds. We have expressed the Arabidopsis 1-Cys Prx AtPER1 in Escherichia coli and show that this protein has antioxidant activity in vitro and protects E. coli in vivo against the toxic oxidant cumene hydroperoxide. Although some 1-Cys Prxs are targeted to the nucleus, a green fluorescent protein-AtPER1 fusion protein was also localized to the cytoplasm in an onion epidermis subcellular localization assay. It has been proposed that seed Prxs are involved in maintenance of dormancy and/or protect the embryo and aleurone layer surviving desiccation against damage caused by reactive oxygen species. These hypotheses were tested using transgenic Arabidopsis lines overexpressing the barley (Hordeum vulgare) 1-Cys PER1 protein and lines with reduced levels of AtPER1 due to antisensing or RNA interference. We found no correlation between Prx levels and the duration of the after-ripening period required before germination. Thus, Prxs are unlikely to contribute to maintenance of dormancy. RNA interference lines almost devoid of AtPER1 protein developed and germinated normally under standard growth room conditions. However, seeds from lines overexpressing PER1 were less inclined to germinate than wild-type seeds in the presence of NaCl, mannitol, and methyl viologen, suggesting that Prx can sense harsh environmental surroundings and play a part in the inhibition of germination under unfavorable conditions. PMID- 14526117 TI - Overexpression of the FRO2 ferric chelate reductase confers tolerance to growth on low iron and uncovers posttranscriptional control. AB - The Arabidopsis FRO2 gene encodes the low-iron-inducible ferric chelate reductase responsible for reduction of iron at the root surface. Here, we report that FRO2 and IRT1, the major transporter responsible for high-affinity iron uptake from the soil, are coordinately regulated at both the transcriptional and posttranscriptional levels. FRO2 and IRT1 are induced together following the imposition of iron starvation and are coordinately repressed following iron resupply. Steady-state mRNA levels of FRO2 and IRT1 are also coordinately regulated by zinc and cadmium. Like IRT1, FRO2 mRNA is detected in the epidermal cells of roots, consistent with its proposed role in iron uptake from the soil. FRO2 mRNA is detected at high levels in the roots and shoots of 35S-FRO2 transgenic plants. However, ferric chelate reductase activity is only elevated in the 35S-FRO2 plants under conditions of iron deficiency, indicating that FRO2 is subject to posttranscriptional regulation, as shown previously for IRT1. Finally, the 35S-FRO2 plants grow better on low iron as compared with wild-type plants, supporting the idea that reduction of ferric iron to ferrous iron is the rate limiting step in iron uptake. PMID- 14526118 TI - Pathogen-responsive expression of a putative ATP-binding cassette transporter gene conferring resistance to the diterpenoid sclareol is regulated by multiple defense signaling pathways in Arabidopsis. AB - The ATP-binding cassette (ABC) transporters are encoded by large gene families in plants. Although these proteins are potentially involved in a number of diverse plant processes, currently, very little is known about their actual functions. In this paper, through a cDNA microarray screening of anonymous cDNA clones from a subtractive library, we identified an Arabidopsis gene (AtPDR12) putatively encoding a member of the pleiotropic drug resistance (PDR) subfamily of ABC transporters. AtPDR12 displayed distinct induction profiles after inoculation of plants with compatible and incompatible fungal pathogens and treatments with salicylic acid, ethylene, or methyl jasmonate. Analysis of AtPDR12 expression in a number of Arabidopsis defense signaling mutants further revealed that salicylic acid accumulation, NPR1 function, and sensitivity to jasmonates and ethylene were all required for pathogen-responsive expression of AtPDR12. Germination assays using seeds from an AtPDR12 insertion line in the presence of sclareol resulted in lower germination rates and much stronger inhibition of root elongation in the AtPDR12 insertion line than in wild-type plants. These results suggest that AtPDR12 may be functionally related to the previously identified ABC transporters SpTUR2 and NpABC1, which transport sclareol. Our data also point to a potential role for terpenoids in the Arabidopsis defensive armory. PMID- 14526119 TI - Transport of the two natural auxins, indole-3-butyric acid and indole-3-acetic acid, in Arabidopsis. AB - Polar transport of the natural auxin indole-3-acetic acid (IAA) is important in a number of plant developmental processes. However, few studies have investigated the polar transport of other endogenous auxins, such as indole-3-butyric acid (IBA), in Arabidopsis. This study details the similarities and differences between IBA and IAA transport in several tissues of Arabidopsis. In the inflorescence axis, no significant IBA movement was detected, whereas IAA is transported in a basipetal direction from the meristem tip. In young seedlings, both IBA and IAA were transported only in a basipetal direction in the hypocotyl. In roots, both auxins moved in two distinct polarities and in specific tissues. The kinetics of IBA and IAA transport appear similar, with transport rates of 8 to 10 mm per hour. In addition, IBA transport, like IAA transport, is saturable at high concentrations of auxin, suggesting that IBA transport is protein mediated. Interestingly, IAA efflux inhibitors and mutations in genes encoding putative IAA transport proteins reduce IAA transport but do not alter IBA movement, suggesting that different auxin transport protein complexes are likely to mediate IBA and IAA transport. Finally, the physiological effects of IBA and IAA on hypocotyl elongation under several light conditions were examined and analyzed in the context of the differences in IBA and IAA transport. Together, these results present a detailed picture of IBA transport and provide the basis for a better understanding of the transport of these two endogenous auxins. PMID- 14526120 TI - Starch-branching enzyme I-deficient mutation specifically affects the structure and properties of starch in rice endosperm. AB - We have isolated a starch mutant that was deficient in starch-branching enzyme I (BEI) from the endosperm mutant stocks of rice (Oryza sativa) induced by the treatment of fertilized egg cells with N-methyl-N-nitrosourea. The deficiency of BEI in this mutant was controlled by a single recessive gene, tentatively designated as starch-branching enzyme mutant 1 (sbe1). The mutant endosperm exhibited the normal phenotype and contained the same amount of starch as the wild type. However, the mutation apparently altered the fine structure of amylopectin. The mutant amylopectin was characterized by significant decrease in both long chains with degree of polymerization (DP) > or = 37 and short chains with DP 12 to 21, marked increase in short chains with DP < or = 10 (A chains), and slight increase in intermediate chains with DP 24 to 34, suggesting that BEI specifically synthesizes B1 and B2-3 chains. The endosperm starch from the sbe1 mutant had a lower onset concentration for urea gelatinization and a lower onset temperature for thermo-gelatinization compared with the wild type, indicating that the genetic modification of amylopectin fine structure is responsible for changes in physicochemical properties of sbe1 starch. PMID- 14526121 TI - De novo synthesis of PCR templates for the development of SARS diagnostic assay. AB - A novel coronavirus was identified as the cause for severe acute respiratory syndrome (SARS). The complete sequence of SARS genome has provided an opportunity for the development of molecular diagnostic assays. To restrain further outbreak of SARS, the World Health Organization has posted several pairs of polymerase chain reaction (PCR) primers for early diagnosis and urged more research to be done on PCR protocols. Here we report a strategy for the de novo synthesis of PCR templates complimentary to the SARS virus genome, which has the advantage of working on PCR templates without concern about viral infection and also has the advantage that it can be used by those who do not have access to the SARS virus. This highly efficient and safe strategy for obtaining SARS gene fragments is useful for the development of PCR assays, as well as for the preparation of reliable positive controls for PCR testing kits. PMID- 14526123 TI - Separation of spermatogenic cells from adult transgenic mouse testes using unit gravity sedimentation. AB - During the final stage of spermatogenesis (i.e., spermiogenesis), round spermatids differentiate into mature spermatozoa. This transformation is mediated by a suite of nuclear packaging proteins. These include the transition proteins and the protamines. The two human protamines PRM1 and PRM2, and transition protein TNP2, are encoded by a single chromatin domain bounded by two regions of matrix attachment. Previous transgenic studies in our laboratory have shown that mice harboring a 40-kb segment of human chromosome 16p13.13 containing the PRM1- > PRM2-->TNP2 domain express the transgene in a haploid-specific, copy number dependent, and position-independent manner. While these results indicate that this segment of the genome is a complete structural and functional regulatory unit, the elements governing the haploid expression of this suite of genes remain to be clearly defined. The preparation of spermatogenic cells is required to begin to address this mechanism. The CELSEP (Wescor/Dupont Inc. Wilmington, DE) unit-gravity sedimentation apparatus provides a simple, efficient, and reproducible means to separate testicular germ cells at all stages along this differentiative pathway. The high quality and integrity of germ cells obtained by this means provides a valuable resource for characterizing the molecular mechanisms governing the regulation of the PRM1-->PRM2-->TNP2 domain during spermatogenesis. A discussion of the CELSEP apparatus and the application of this methodology in our laboratory are presented. PMID- 14526122 TI - Detection of mutations in RET proto-oncogene codon 634 through double tandem hybridization. AB - We developed a procedure to detect the 7 point mutations at Cys634 of the proto oncogene RET, which is responsible for medullary thyroid carcinoma (MTC). Genomic DNA was prepared from blood samples obtained from normal and MTC-affected individuals belonging to a family with a history of the disease. The RET genotype for each individual was first established by performing restriction and sequencing analyses. Single-stranded target DNA was prepared by asymmetric polymerase chain reaction (PCR) amplification of a 93-bp fragment containing Cys634. The target was annealed with pairs of prelabeled stacking oligonucleotides designed to create appropriate 7-nucleotide gaps, which served as the sites of subsequent hybridization with glass-immobilized 7-mer probes. The target-stacking oligonucleotide duplexes were hybridized with DNA chips containing a set of eight 7-mer probes designed to detect the wild-type sequence and the seven point mutations described. We tested two sets of immobilized probes containing internal or 5'-terminal codon-634 single-base variations. Both groups of probes were able to discriminatively identify the mutations. The hybridization patterns indicated that the disease in this family was due to the C634Y mutation, in accord with the original sequence analysis. The hybridization-based mutation assignment was additionally supported by determination of the control homozygous and heterozygous hybridization patterns produced with synthetic targets having the normal or codon 634 mutant sequences. The effects of mismatch type and nearest-neighbor sequences on the occurrence of false-positive (mismatched) hybridizations are discussed. PMID- 14526124 TI - Gene therapy for lung cancer. AB - Lung cancer continues to be the largest killer of Americans due to cancer. Although progress has been made, with advances in chemotherapy, the majority of patients diagnosed with lung cancer ultimately succumb to the disease. A better understanding of the molecular pathogenesis of lung cancer is demonstrating how alterations in oncogenes and tumor suppressor genes control lung cancer initiation, growth, and survival. In this article, attempts to target molecular alterations in lung cancer using gene therapy techniques are reviewed. These include introducing suicide genes into tumor cells, replacement of defective tumor suppressor genes, inactivating oncogenes, and immunotherapy-based approaches using gene therapy technology. The major barrier for these techniques continues to be the inability to specifically target tumor cells while sparing normal cells. Nonetheless, these approaches are likely to yield important biologic and clinical data which will further the progress of lung cancer treatment. PMID- 14526127 TI - The race from chronic pancreatitis to pancreatic cancer. PMID- 14526126 TI - Therapeutic inhibition of angiogenesis. AB - Angiogenesis, or the growth of new blood vessels, has, in recent years, become an area of intense scientific research. The primary reason for this has been the realization that angiogenesis plays a key role in many common pathologies, and that its inhibition could have profound implications in the treatment of these disorders. A substantial number of anti-angiogenic agents have now been identified; however, none has, as of yet, achieved widespread acceptance in the clinic. Many agents have been identified as the result of clearly defined research programs, such as the inhibitors of the vascular endothelial growth factor transmembrane tyrosine kinase receptors, but many other simply by screening. The purpose of this article is to review the wealth of information available on known anti-angiogenic agents and to assess their future potential. PMID- 14526128 TI - CFTR, PRSS1 and SPINK1 mutations in the development of pancreatitis in Brazilian patients. AB - CONTEXT: Mutations in cystic fibrosis transmembrane conductance regulator (CFTR), in cationic trypsinogen (PRSS1) and in serine protease inhibitor Kazal type 1 (SPINK1) genes have been associated with chronic pancreatitis (alcohol related, idiopathic and hereditary). However, the inheritance pattern is still not clear. PATIENTS: Eighty-two unrelated Brazilian patients with chronic pancreatitis (alcohol-related disease in 64, idiopathic disease in 16, and hereditary disease in 2). Two hundred unrelated individuals with an ethnic distribution comparable to the patients were studied as controls. MAIN OUTCOME MEASURE: Detection of mutations in CFTR, PRSS1, and SPINK1 genes. RESULTS: Mutations in the CFTR gene were found in 8 patients (9.8%) with chronic pancreatitis, 5 of them with idiopathic disease. Interestingly, the only clinical symptom in a male patient in the alcoholic group, who was a compound heterozygote (DeltaF508/R170C) for two CFTR mutations, was pancreatitis without infertility or pulmonary involvement. In the PRSS1 gene, the E79K change in exon 3 was found in one patient (1.2%) with alcohol-related chronic pancreatitis. Four different alterations were identified in the SPINK1 gene. CONCLUSIONS: Mutations in the CFTR gene represent the major cause of idiopathic chronic pancreatitis in Brazilian patients. No mutation was found in the PRSS1 gene among our patients suggesting further genetic heterogeneity for hereditary and idiopathic chronic pancreatitis. Interestingly, the most frequent SPINK1 N34S mutation was not present in patients or controls. Moreover, the -253C allele for the SPINK1 gene was significantly more frequent in patients than controls (P=0.004), suggesting that it might represent a risk factor for the development of pancreatitis in our population. PMID- 14526129 TI - Functional disturbance of biliary indocyanine green excretion in rat cerulein pancreatitis followed by endotoxemia: role of the prime and the second attack. AB - CONTEXT: Hepatic injury is considered one of the critical complications associated with acute pancreatitis. It was proposed that initial insults to the liver in the early phase of the attack have an important priming effect, and the subsequent infectious attack (e.g. infectious pancreatic necrosis, bacterial translocation episode) constitutes a second attack on the liver. OBJECTIVE: To evaluate the role of priming by induction of cerulein pancreatitis and a following second attack by endotoxemia. MAIN OUTCOME MEASURES: Plasma clearance and biliary excretion of indocyanine green (a hepatophillic hydrophobic organic anion). DESIGN: A model of acute pancreatitis in rats. SETTING: Four groups of rats: untreated control, cerulein pancreatitis, endotoxemia and endotoxemia following the induction of cerulein pancreatitis (pancreatitis + endotoxemia). RESULTS: Biliary indocyanine green excretion was significantly disturbed only in the pancreatitis + endotoxemia group. Plasma clearance (a reflection of hepatic uptake) of indocyanine green from the blood was only slightly affected in endotoxemia group. CONCLUSION: Biliary secretion is quite sensitive to this hepatic injury model. Both the preceding priming insult and the following second attack are important in the development of hepatic injury. PMID- 14526130 TI - Can roxithromycin and betamethasone induce acute pancreatitis? A case report. AB - CONTEXT: Acute pancreatitis has been reported in a few cases treated with macrolides or glucocorticoids. CASE REPORT: We report the case of a 58 year old patient who, after 2 days of treatment with roxithromycin and betamethasone, manifested acute pancreatitis. Other causes of the disease were ruled out. No re occurrence of pancreatitis was observed in a 16 month follow-up. CONCLUSION: Our case sheds new light on glucocorticoid pancreatotoxicity and confirms the role of macrolides as potential pancreatotoxic drugs. PMID- 14526125 TI - Genomic imprinting and endosperm development in flowering plants. AB - Genomic imprinting, the parent-of-origin-specific expression of genes, plays an important role in the seed development of flowering plants. As different sets of genes are imprinted and hence silenced in maternal and paternal gametophyte genomes, the contributions of the parental genomes to the offspring are not equal. Imbalance between paternally and maternally imprinted genes, for instance as a result of interploidy crosses, or in seeds in which imprinting has been manipulated, results in aberrant seed development. It is predominantly the endosperm, and not or to a far lesser extent the embryo, that is affected by such imbalance. Deviation from the normal 2m:1p ratio in the endosperm genome has a severe effect on endosperm development, and often leads to seed abortion. Molecular expression data for imprinted genes suggest that genomic imprinting takes place only in the endosperm of the developing seed. Although far from complete, a picture of how imprinting operates in flowering plants has begun to emerge. Imprinted genes on either the maternal or paternal side are marked and silenced in a process involving DNA methylation and chromatin condensation. In addition, on the maternal side, imprinted genes are most probably under control of the polycomb FIS genes. PMID- 14526131 TI - Selective serotonin reuptake inhibitors and suicide: is the evidence, as with beauty, in the eye of the beholder? PMID- 14526132 TI - Major depressive disorder with anger: a bipolar spectrum disorder? AB - BACKGROUND: Depression with anger may be more common in bipolar disorders. The aim of the study was to assess whether major depressive disorder (MDD) with anger could be included in the bipolar spectrum, by comparing it to MDD without anger and to bipolar II disorder. METHODS: Consecutive outpatients (281 bipolar II disorder and 202 MDD) presenting for major depressive episode (MDE) treatment were interviewed with the DSM-IV structured clinical interview. Clinical variables used to support the inclusion of MDD with anger in the bipolar spectrum were age of onset, many MDE recurrences, atypical features of depression, depressive mixed state (an MDE plus some concurrent hypomanic symptoms), and bipolar family history. RESULTS: Frequency of MDE with anger was 50.5% [61.2% in bipolar II, and 35.6% in MDD (z = 5.5, p = 0.0000, 95% CI 16.8-43.3%)]. Logistic regression of MDE with anger (dependent variable) versus bipolar variables showed that MDE with anger was significantly associated with all bipolar variables, apart from recurrences. MDD with anger, compared with MDD without anger, had significantly lower age of onset, more marked depressive mixed state, a bipolar family history with more cases, but comparable atypical features and Global Assessment of Functioning scores. MDD with anger, compared with bipolar II disorder, had significantly higher age of onset, less atypical features, and a bipolar family history with less cases. CONCLUSIONS: MDE with anger was common in outpatients (more in bipolar II disorder). MDD with anger may be midway between MDD without anger and bipolar II disorder, and might be included into the bipolar spectrum. However, MDD with anger does not appear to be associated with the often reported negative response to monotherapy with antidepressants. PMID- 14526133 TI - Alexithymia, depression and social support among Japanese workers. AB - BACKGROUND: A number of studies have shown that social support has a direct beneficial effect on well-being and also serves as a buffer to protect people from health problems due to excessive stress. Although preliminary studies report a positive relationship of alexithymia both with depression and reduced social support, there is no study examining whether the beneficial effect of social support on depression differs with the presence of alexithymia. METHODS: A total of 120 workers aged 19-39 completed the 20-item Toronto Alexithymia Scale (TAS 20) to measure alexithymia, the Beck Depression Inventory-II (BDI-II) to evaluate depressive symptomatology, and the Job Content Questionnaire (JCQ) to assess job strain based on Karasek's demand-control-support model. The interrelationship among TAS-20, BDI-II and 3 subscales of JCQ (job demand, control, and support) were examined. RESULTS: A significant association of depression with low support and high alexithymia was observed. Alexithymia was also associated with reduced support. Further, a statistically significant interaction between alexithymia and support in terms of their effect on depression was observed. Nonalexithymic individuals with low support showed a significantly higher depression score than those who received high support, while alexithymics did not differ in their depression score depending on the degree of support. Consistent results were obtained from the logistic regression analysis examining the odds ratio for depression by support by alexithymia; a significantly increased odds ratio for depression associated with low social support was observed only among nonalexithymics. CONCLUSIONS: Alexithymic individuals might be unable to benefit from social support because of their cognitive deficits of emotion. PMID- 14526134 TI - The use of defenses and physician health care costs: are physician health care costs lower in persons with more adaptive defense profiles? AB - BACKGROUND: The objective of the present study was to determine if persons who use more adaptive defenses have lower physician health care costs compared to those who use less adaptive defenses. METHODS: We randomly selected 667 persons from the 1995 population-based Nova Scotia Health Survey who completed a videotaped structured interview. Each interview was rated for typical defense use by the Defense-Q. We obtained physician health care costs for 3 months before and after the interview, as well as medical diagnoses and measures of psychological functioning. RESULTS: A more adaptive defense profile significantly predicted lower future physician health care costs. These results were found when controlling for other psychosocial variables, before and after controlling for previous physician health care costs, and when testing only within a physically healthy subsample. Results of secondary analyses showed that a more adaptive defense profile was positively related to a number of psychosocial variables, such as nurse's rating of competence, lack of depressive symptoms, and days at work. CONCLUSIONS: The adaptiveness of a person's defense use in managing affect is important in predicting physician health care costs as well as psychosocial functioning. PMID- 14526135 TI - Ageing styles: subjective well-being and somatic complaints in inpatients aged >/=60 years. AB - BACKGROUND: Recent gerontological research shows a high variability in the elderly population. The aim of the present study is to investigate by means of cluster analysis, as to whether different 'ageing styles' can be identified in a sample of cognitively not impaired older persons, and to assess discrepancies between self-rated and expert-rated psychological and physical health. METHOD: 243 patients aged 60 years and older in a general hospital were investigated using a clinical psychiatric interview, expert ratings, and self-report scales after extensive internal medical diagnostic evaluation. RESULTS: Five clusters, representing significantly different 'ageing styles' with regard to subjective well-being and subjective physical complaints, could be identified. Two clusters, which together constituted 57% of the sample, represented quite positive and desirable ageing styles. In two clusters (altogether 23% of the sample), there was a discrepancy between medical findings and subjective physical complaints. CONCLUSIONS: These findings support the notion of high variability in the elderly and the existence of subgroups, which show a discrepancy between subjective assessment of well-being and expert assessment of psychological and physical health and impairment. The recording of subjectively perceived physical complaints should, therefore, be part of the evaluation of older peoples' subjective well-being. PMID- 14526136 TI - Prevalence and risk factors of posttraumatic stress disorder in older adults. AB - BACKGROUND: Posttraumatic stress disorder (PTSD) has scarcely been researched in the elderly. There is no population-based information on prevalence and risk factors in older persons. Patients with PTSD are often not recognized or incorrectly diagnosed. As the disorder has great implications for the quality of life, a correct diagnosis and treatment are crucial. Increased knowledge on vulnerability factors for PTSD can facilitate diagnostic procedures and health management in the elderly. METHODS: PTSD cases were found following a two-phase sampling procedure: a random selection of 1,721 subjects were screened and in 422 subjects a psychiatric diagnostic interview was administered. Prevalence of PTSD and subthreshold PTSD were calculated. Vulnerability factors regarding demographics, physical health, personality, social factors, recent distress and adverse events in early childhood were assessed. RESULTS: 6-month prevalence of PTSD and of subthreshold PTSD was 0.9 and 13.1%, respectively. The strongest vulnerability factors for both PTSD and subthreshold PTSD were neuroticism and adverse events in early childhood. CONCLUSIONS: This is the first population based study on PTSD in older persons. With a 6-month prevalence of almost 1% the disease is not rare. Comparisons with younger populations suggest some accumulation of cases among older people reflecting the chronic risk factors, which are found in this study: neuroticism and adverse events in early childhood. PMID- 14526137 TI - Psychological assessment in cardiac rehabilitation. AB - BACKGROUND: While there has been an upsurge of interest in the psychiatric correlates of myocardial infarction, little is known about the presence of psychological distress in the setting of cardiac rehabilitation. METHODS: A consecutive series of 61 patients with recent myocardial infarction who participated in a cardiac rehabilitation program was evaluated by means of both observer-rated (DSM and DCPR) and self-rated (Psychosocial Index) methods. A follow-up of this patient population was undertaken (median = 2 years). Survival analysis was used to characterize the clinical course of patients. RESULTS: Twenty percent of patients had a DSM-IV diagnosis (in half of the cases minor depression). An additional 30% of patients presented with a DCPR cluster, such as type A behavior and irritable mood. Only high levels of self-perceived stressful life circumstances and psychological distress approached statistical significance as a psychological risk factor for cardiovascular events after myocardial infarction. CONCLUSIONS: Psychological evaluation of patients undergoing cardiac rehabilitation needs to incorporate both clinical (DSM) and subclinical (DCPR) methods of classification. Type A behavior was present in about a quarter of patients and can be studied in specific subgroups of cardiovascular patients defined by DCPR. PMID- 14526138 TI - Principles of nidotherapy in the treatment of persistent mental and personality disorders. AB - BACKGROUND: The principles of a psychological intervention for psychiatric disorders, nidotherapy, are described following its use in the treatment of patients with severe mental illness, most of whom also have personality disorders. Nidotherapy is the collaborative systematic assessment and modification of the environment to minimise the impact of any form of mental disorder on the individual or on society. It is particularly appropriate to consider for chronic and recurring disorders in which there is no prospect of short-term improvement. METHODS: An analysis of the treatment suggests its main tenets are the development of a joint consensus (collateral collocation) of environmental change needed, formulation of targets, consequent monitoring and adjustments made to test the effectiveness of intervention, and a joint structure (arbitrage solving) to adjudicate when changes are considered feasible or impractical by one party and not the other. RESULTS: Case vignettes are used to describe the main features of the treatment. Although nidotherapy includes some of the elements of cognitive, occupational and family therapy, and even social work, it is different in that it focuses on changing the environment, not the person. It is therefore much more acceptable to those patients who have repeatedly challenged the justification of conventional therapy for their conditions. CONCLUSIONS: Nidotherapy deserves further enquiry as a useful addition to existing psychological treatments. PMID- 14526139 TI - Do SSRIs increase the risk of suicide among depressives even if they are taking only placebo? PMID- 14526140 TI - Do SSRIs cause suicide? PMID- 14526142 TI - Prescribing cyclic antidepressants for vitiligo patients: which agents are superior, which are not? PMID- 14526143 TI - First steps in the assessment of cognitive-emotional organisation within the framework of Guidano's model of the self. PMID- 14526144 TI - Should patients with locally advanced, non-metastatic carcinoma of the pancreas be irradiated? AB - A small number of patients exist with carcinoma of the pancreas with an inoperable but not metastasized tumor. Prospective randomized studies defined the standard of combined radiochemotherapy during the early 1980s for these patients. Since then, new drugs have shown considerable activity and in parallel improvements in radiotherapy treatment planning and delivery have been achieved. Therefore, it is time to ask whether patients with locally advanced, inoperable pancreatic cancer without metastases should still be irradiated or not. This review summarizes the current literature on combined radiochemotherapy for locally advanced carcinoma of the pancreas. Median survival times of 10-11 months and 1-year survival rates of about 40% can be achieved with modern radiochemotherapy regimens. PMID- 14526145 TI - Synergistic activity of gamma-linolenic acid and cytotoxic drugs against pancreatic adenocarcinoma cell lines. AB - BACKGROUND: Gamma-linolenic acid (GLA) is growth inhibitory both in vitro and in vivo, at doses non-toxic to non-cancer cells. Chemotherapeutic agents have limited activity in pancreatic cancer. Interactions between GLA and cytotoxic drugs have not previously been investigated; any synergy might improve the therapeutic effect of these agents. AIM: To investigate possible interactions between GLA and 5-fluorouracil (5-FU) or gemcitabine against pancreatic cancer cell lines in vitro. METHODS: Two pancreatic cancer cell lines were exposed to GLA alone and in combination with 5-FU or gemcitabine. Residual viable biomass was measured using the MTT assay and the results analysed by the median effect method of Chou and Talalay [Adv Enzyme Regul 1984;22:27-55]. RESULTS: GLA concentrations of 3.9- 125 microg/ml had a synergistic or additive growth inhibitory effect on all tested concentrations of gemcitabine. Synergism was demonstrated between GLA and 5-FU only at concentrations of 62.5-125 microg/ml of 5-FU. CONCLUSION: GLA has a synergistic effect with gemcitabine at concentrations that correspond to in vivo therapeutic doses. GLA with 5-FU is synergistic only at a tight range of high concentrations of 5-FU. GLA lacks toxic side effects and may be useful in combination with gemcitabine. PMID- 14526146 TI - Elevated protein carbonyls as plasma markers of oxidative stress in acute pancreatitis. AB - BACKGROUND: Experimental studies have demonstrated that protein and lipid oxidation is a feature of acute pancreatitis and that antioxidant pretreatment can ameliorate the severity of the disease. Justification for a clinical trial of antioxidant therapy requires stronger evidence for oxidative stress in patients. AIMS: To determine if oxidative stress is evident in patients with acute pancreatitis on admission to hospital, if it increases after admission and if it is related to disease severity. METHODS: Measurement of plasma concentrations of protein carbonyls and malondialdehyde as markers of protein oxidation and lipid peroxidation, respectively, in a consecutive series of 85 patients with acute pancreatitis 0, 2 and 5 days after admission. RESULTS: Patients with acute pancreatitis had significantly increased concentrations of protein carbonyls in plasma on recruitment (median 27 h after the onset of symptoms) that persisted over 5 days. Protein carbonyls were higher in severe compared with mild disease (median 0.099 and 0.043 nmol/mg protein, respectively, p = 0.0016). They were higher at day 0 in patients recruited with more established pancreatitis than in those presenting early. No increases in malondialdehyde were seen. Receiver operator characteristic curve analysis demonstrated that protein carbonyls at day 0 were comparable with C-reactive protein at predicting pancreatitis severity. CONCLUSION: Our demonstration of substantial protein oxidation provides further evidence for oxidative stress in patients with severe pancreatitis. Our results suggest that there could be a window for early antioxidant intervention and that protein carbonyls could be a useful plasma marker of oxidative injury. PMID- 14526147 TI - Is late antibiotic prophylaxis effective in the prevention of secondary pancreatic infection? AB - BACKGROUND: Secondary infection of the inflamed pancreas is the principal cause of death after severe acute pancreatitis (AP). Although patients are not always managed early in the course of AP in clinical practice, prophylactic antibiotics that were used in experimental studies in rats were always initiated early after induction of pancreatitis. The effectiveness of antibiotics initiated later is unknown. AIM: The aim of this study was to compare the effectiveness of ciprofloxacin and meropenem initiated early versus later in the course of acute necrotizing pancreatitis (ANP) in rats. METHODS: 100 Sprague-Dawley rats were studied. ANP was induced in rats by intraductal injection of 3% taurocholate. Rats were divided randomly into five groups: group I rats received normal saline as a placebo, group II and IV rats received three times daily meropenem 60 mg/kg i.p. at 2 and 24 h, respectively and group III and V rats received twice daily ciprofloxacin 50 mg/kg i.p. at 2 and 24 h, respectively, after induction. At 96 h, all rats were killed for quantitative bacteriologic study. A point-scoring system of histological features was used to evaluate the severity of pancreatitis. RESULTS: Meropenem and ciprofloxacin initiated 2 h after induction of pancreatitis significantly reduced the prevalence of pancreatic infection (p < 0.001 and p < 0.04, respectively) as compared to controls. Neither of the antibiotics initiated later during the course of AP caused a significant decrease in pancreatic infection in rats (p > 0.05). Although the rats treated early infected less frequently than the rats treated later, the comparison reached statistical significance only in the meropenem group (p < 0.02). CONCLUSION: Early antibiotic treatment reduces pancreatic infection more efficiently than late antibiotic treatment in ANP in rats. PMID- 14526148 TI - The Lundh test and faecal elastase 1 determination in chronic pancreatitis: a comparative study. AB - BACKGROUND/AIM: A reduced exocrine pancreatic function supports the diagnosis of chronic pancreatitis (CP) in symptomatic patients. A sensitive test for a reduced exocrine function is decisive, especially when morphological changes are missing. The aim of this study was to compare the indirect faecal elastase 1 (FE-1) test with the direct Lundh test in patients with and without definite diagnostic imaging findings. METHODS: Eighty-nine patients with clinical signs suggesting CP or having an established diagnosis of CP had a Lundh test and an estimation of FE 1 performed. All patients underwent abdominal ultrasonography and/or computed tomography. RESULTS: A significant correlation (r = 0.70, p < 0.02) was found between FE-1 and meal-stimulated intraduodenal lipase. Using the Lundh test as reference, the predictive values of a positive and negative FE-1 test were for all patients investigated 81 and 73%, respectively. Patients with equivocal imaging findings had lower predictive values (positive predictive value 57%; negative predictive value 71%) as compared with patients with moderate or marked imaging findings (positive predictive value 84%; negative predictive value 78%). Fair to moderate chance-corrected agreement was found between Lundh test and FE-1 concentration. CONCLUSIONS: In patient with imaging findings suggesting CP, FE-1 determination is a highly sensitive test for exocrine pancreatic function, but in patients with equivocal imaging findings, the predictive power of FE-1 limits the test to serve as a reliable diagnostic tool. PMID- 14526149 TI - High prevalence of exocrine pancreatic insufficiency in diabetes mellitus. A multicenter study screening fecal elastase 1 concentrations in 1,021 diabetic patients. AB - BACKGROUND: There have been numerous reports on pancreatic exocrine dysfunction in diabetes mellitus using either direct or indirect function tests. The measurement of fecal elastase 1 concentrations (FEC) has been used as a screening tool for exocrine pancreatic disease in different patient groups indicating a high prevalence of exocrine dysfunction in diabetic populations. In this study we had the opportunity to study more than 1,000 diabetic patients to confirm recent observations in smaller populations. METHODS: FEC were measured by ELISA in 323 patients with type 1 and 697 type 2 diabetes mellitus. Subjects with a history of alcohol abuse, gastrointestinal surgery, cancer or inflammatory diseases were not included. Diabetes history and clinical data were recorded using a standard case report form. FINDINGS: 1,021 patients (334 female, 687 male; mean age 50 years; mean diabetes duration 11 years; mean age at onset of diabetes 39 years) were studied. FEC was normal (>200 microg/g) in 59.3% and severely reduced (<100 microg/g) in 22.9%. There were significant differences between type 1 and type 2 patients as well as between insulin-treated and non-insulin-treated patients. Furthermore, there were weak associations between FEC and diabetes duration, age at onset of diabetes and body mass index, respectively. INTERPRETATION: We could confirm that both type 1 and type 2 diabetic patients show pathological exocrine function in high prevalence. Exocrine insufficiency seems to be correlated to early onset of endocrine failure, long-lasting diabetes mellitus and low body mass index levels. PMID- 14526150 TI - Primary pancreatic lymphoma presenting as acute pancreatitis. AB - Primary pancreatic lymphoma is a rare disease and usually presents with nonspecific symptoms like abdominal pain, weight loss, or jaundice. Here we report a case of primary pancreatic lymphoma presenting as acute pancreatitis. PMID- 14526151 TI - A randomised clinical trial to assess the effect of total enteral and total parenteral nutritional support on metabolic, inflammatory and oxidative markers in patients with predicted severe acute pancreatitis (APACHE II > or =6). AB - BACKGROUND: Total enteral nutrition (TEN) within 48 h of admission has recently been shown to be safe and efficacious as part of the management of severe acute pancreatitis. Our aim was to ascertain the safety of immediate TEN in these patients and the effect of TEN on systemic inflammation, psychological state, oxidative stress, plasma glutamine levels and endotoxaemia. METHODS: Patients admitted with predicted severe acute pancreatitis(APACHE II score >5) were randomised to total enteral (TEN; n = 8) or total parenteral nutrition (TPN; n = 9). Measurements of systemic inflammation (C-reactive protein), fatigue (visual analogue scale), oxidative stress (plasma thiobarbituric acid-reactive substances), plasma glutamine and anti-endotoxin IgG and IgM antibody concentrations were made on admission and repeated on days 3 and 7 thereafter. Clinical progress was monitored using APACHE II score. Organ failure and complications were recorded. RESULTS: All patients tolerated the feeding regime well with few nutrition-related complications. Fatigue improved in both groups but more rapidly in the TEN group. Oxidative stress was high on admission and rose by similar amounts in both groups. Plasma glutamine concentrations did not change significantly in either group. In the TPN group, 3 patients developed respiratory failure and 3 developed non-respiratory single organ failure. There were no such complications in the TEN group. Hospital stay was shorter in the TEN group [7(4-14) vs. 10 (7-26) days; p = 0.05] as was time to passing flatus and time to opening bowels [1 (0-2) vs. 2 (1-5)days; p = 0.01]. The cost of TEN was considerably less than of TPN. CONCLUSION: Immediate institution of nutritional support in the form of TEN is safe in predicted severe acute pancreatitis. It is as safe and as efficacious as TPN and may be beneficial in the clinical course of this disease. PMID- 14526153 TI - Reversal of diabetes in the rat by injection of hematopoietic stem cells infected with recombinant adeno-associated virus containing the preproinsulin II gene. AB - AIM: To study the effect of injecting hematopoietic stem cells containing the preproinsulin gene II (rI2) via recombinant adeno-associated virus (rAAV) into normal and streptozotocin-diabetic rats. METHODS: rI2 was transfected into rat hematopoietic stem cells using rAAV vector. Stem cells were injected by intravenous route into normal and STZ-induced diabetic rats to study blood sugar and expression of rI2 in various tissues. The pLP-1 recombinant plasmid containing rI2 (vLP-1) was engineered as previously described. Bone marrow from female Wistar-Furth rats was enriched for stem cells by using plastic adherence and monoclonal antirat CD3 and CD45 RA to deplete T and B cells. The remaining cells were exposed to vLP-1 (multiplicity of infection MOI =50:1 or 100:1) for 2 h. Approximately ten million exposed stem cells were injected by intravenous route into each animal; there were four groups: normal animals at MOI 50:1 (group 1) or MOI 100:1 (group 2); group 3 animals (n = 9) were streptozotocin-induced diabetic animals at MOI 100:1. Animals that showed reversal of diabetes from group 3 were sacrificed for study of gene expression at weeks 1, 2, and 6, respectively. Control diabetic animals did not receive stem cells or virus constituted group 4. Expression of rI2 was analyzed by RT-PCR and Southern analyses. RESULTS: Despite introduction of insulin gene, groups 1 and 2 had blood sugar concentrations that remained within normal levels, while 3 of 9 animals in group 3 showed reversal of diabetes; using RT-PCR,group 1 expressed rI2 in liver, spleen, thymus, brain, and heart at week 1 only. In group 2, rI2 was seen in the thymus up to 6 weeks; in diabetic animals (group 3) rI2 was seen in liver, bone marrow, spleen, thymus, and peripheral blood lymphocytes at week 2 and in thymus and lymphocytes at week 6. CONCLUSIONS: We have shown that (1) rAAV is a useful vector for transferring rI2 into rat hematopoietic stem cells; (2) normal animals remained euglycemic after injection of stem cells containing rI2 despite identification in various tissues suggesting autoregulation, and (3) short-term reversal of diabetes was achieved in some animals by injection of stem cells containing rI2. PMID- 14526152 TI - Pro-inflammatory cytokine-induced matrix metalloproteinase-1 (MMP-1) secretion in human pancreatic periacinar myofibroblasts. AB - Matrix metalloproteinases (MMPs) are the proteases involved in the degradation of the extracellular matrix. MMP-1 is thought to be one of the key enzymes in fibrolysis, a process closely related to tissue remodeling. In the present study, we investigated MMP-1 secretion from human pancreatic periacinar myofibroblasts in response to pro-inflammatory cytokines IL-1beta and TNF-alpha. We also attempted to clarify the intracellular signaling pathways mediating the cytokine induced MMP-1 secretion. MMP-1 secretion was measured by an enzyme-linked immunosorbent assay. MMP-1 molecules were analyzed by Western blotting. MMP-1 mRNA expression was evaluated by Northern blotting. IL-1l and TNF-alpha stimulated the MMP-1 secretion in a dose- and time-dependent manner. Ninety percent of MMP-1 was secreted as inactive form (pro-MMP-1). The effects of IL 1beta and TNF-alpha were significantly inhibited by PD98059 MEK/ERK inhibitor). In contrast, SB203580 (p38 MAPK inhibitor), GF109203X (PKC inhibitor), and PDTC (NF-kappaB inhibitor) did not alter the MMP-1 secretion induced by IL-1beta and TNF-alpha. These effects were also observed at them RNA level. In conclusion, in human pancreatic periacinar myofibroblasts, MMP-1 secretion was regulated by the pro-inflammatory cytokines via the MEK/ERK cascade. Thus, human pancreatic periacinar myofibroblasts may play an important role in the remodeling of damaged pancreatic tissue in chronic pancreatitis via MMP-1 secretion. PMID- 14526154 TI - [New developments in hormone replacement therapy]. AB - Hormone replacement therapy with estrogen and progesterone does not appear to protect against Alzheimer's disease and does not improve cognitive functioning. Long-term administration of estrogen as monotherapy may be associated with an increased risk of ovarian cancer. PMID- 14526155 TI - [Gender stereotypes in gynecology and obstetrics: obstacles for young male physicians?]. AB - Up to the mid 1990s, a 'feminization' of medicine took place that was especially pronounced in the field of gynecology. The present paper looks into the reasons for this change based on the literature and results from a study conducted by the author. In 2001, two thirds of the new qualifications in gynecology/obstetrics were awarded to women. The feminist movement has led to the emancipation of women and the assertion that women should be treated by female doctors only. Parallel to this development, gender roles also underwent a change: nowadays, the ideal woman or man is 'androgynous'. Young physicians are both highly instrumental and highly expressive. The increasing similarities between the sexes exert an influence on lifestyle and biographical planning. An increasing number of young physicians no longer prioritize their profession. Male physicians with high expressiveness are often family oriented and consciously refrain from choosing to specialize in gynecology/obstetrics because of the long hours and heavy workload. On the other hand, quite a few women physicians are more instrumental and prioritize their professional commitment. If more male physicians are not attracted into gynecology/obstetrics over the next few years, a process of 'horizontal segregation' could occur, with women physicians accomplishing the patient-focused work while their male colleagues take over the surgery and scientific part of the specialty. It is the very differences between the sexes, however, and the exchange between them, which allow a medical discipline to thrive. PMID- 14526156 TI - [The learning curve in the context of the cesarean section]. AB - BACKGROUND: We created learning curves to define an objective figure according to which residents are capable of performing a cesarean section responsibly without supervision. METHODS: We established learning curves of 9 different outcome variables related to cesarean section for all trainees of obstetrics-gynecology in the years 1995-1999 (blood loss, length of stay, Apgar score, umbilical cord pH value, induction-delivery time, uterotomy-delivery time and total operation time). The change from the steep to the flat part of the curve was determined visually and defined as the end of the initial learning phase. RESULTS: 15 residents performed a total of 371 sections. Concerning the operation time and the induction-delivery time, the steep part of the curve was exceeded after 20 sections. For the 1-min Apgar score and the length of stay, only a slight change in the curve was present; for the remaining parameters, no learning process could be demonstrated. CONCLUSION: For cesarean sections of average risk, 20 procedures are adequate to provide sufficient safety for the patient. PMID- 14526157 TI - [Asymptomatic uterine rupture after two spontaneous vaginal deliveries following prior cesarean section]. AB - We present the case of an almost asymptomatic uterine rupture after two consecutive vaginal deliveries after a prior cesarean delivery (first child). Uterine rupture is rare even after a prior cesarean delivery, permitting a trial of labor after a cesarean delivery in spite of the increased risk of uterine rupture. Vaginal birth after previous cesarean delivery, however, demands a cautious approach. Appropriate recommendations have been published by the ACOG and the 'AG fOr fetomaternale Medizin' (a branch of the German Society of Obstetrics and Gynaecology). In our case diagnosis was made by an vaginal ultrasound examination 7.5 weeks after an uncomplicated vaginal delivery. An additional MRI examination did not result in substantial extra information. For that reason it will be indicated only in exceptional cases. To answer the question whether a vaginal ultrasound examination should routinely be offered after an uncomplicated vaginal delivery with a prior cesarean delivery in the history to preclude uterine rupture further studies are necessary. PMID- 14526158 TI - Efficacy of Acupuncture for the Treatment of Primary Dysmenorrhea. AB - OBJECTIVE: The aim of this study was to evaluate the effect of acupuncture (AP) in the treatment of primary dysmenorrhea (PD). METHODS: A clinical prospective, placebo-controlled trial included 57 women with PD. Of these, 30 were treated with manual AP points: Du 20 (Baihui), bilateral Li 4 (Hegu), Ren 3 (Zhongji), Ren 4 (Guanyuan), Ren 6 (Qihai), bilateral Gb 34 (Yanglingquan), bilateral Ub 23 (Shenshu), bilateral Lp 6 (Sanyinjao) and auriculoacupuncture points (Shenmen); 27 women were treated with placebo AP. AP treatments were considered successful if PD did not occur any more, medication of PD became unnecessary or PD symptoms did not occur for 2 years after the AP treatment. RESULTS: The occurrence of PD in nulliparae was statistically relevant (p < 0.001). Statistically relevant was also the decrease in medication in women to whom AP had been applied (p < 0.0001), which was not the case in the placebo group (p > 0.5). CONCLUSIONS: The success rate of AP for the treatment of PD symptoms within 1 year after the AP treatment is 93.3% in the first group and 3.7% in the placebo group. PMID- 14526159 TI - [Extrajudicial medicolegal opinions]. AB - Patients who endured complications due to a presumed or genuine medical error often request an extrajudicial medicolegal opinion. Frequently, the problems appear to originate from an insufficient (preoperative) information of the patient. These expert opinions are an instructive source of learning about possibly avoidable errors. Publishing such cases, in a completely anonymized form, should hopefully contribute to the prevention of the recurrence of certain mistakes. We present and comment on two postoperative events: (1) incisional hernias after abdominal hysterectomy and (2) a sub-diaphragmatic abscess after laparoscopic removal of a dermoid ovarian cyst with intraabdominal spillage of its content. PMID- 14526162 TI - Trinucleotide repeat instability: a hairpin curve at the crossroads of replication, recombination, and repair. AB - The trinucleotide repeats that expand to cause human disease form hairpin structures in vitro that are proposed to be the major source of their genetic instability in vivo. If a replication fork is a train speeding along a track of double-stranded DNA, the trinucleotide repeats are a hairpin curve in the track. Experiments have demonstrated that the train can become derailed at the hairpin curve, resulting in significant damage to the track. Repair of the track often results in contractions and expansions of track length. In this review we introduce the in vitro evidence for why CTG/CAG and CCG/CGG repeats are inherently unstable and discuss how experiments in model organisms have implicated the replication, recombination and repair machinery as contributors to trinucleotide repeat instability in vivo. PMID- 14526163 TI - The contribution of cis-elements to disease-associated repeat instability: clinical and experimental evidence. AB - Alterations in the length (instability) of gene-specific microsatellites and minisatellites are associated with at least 35 human diseases. This review will discuss the various cis-elements that contribute to repeat instability, primarily through examination of the most abundant disease-associated repetitive element, trinucleotide repeats. For the purpose of this review, we define cis-elements to include the sequence of the repeat units, the length and purity of the repeat tracts, the sequences flanking the repeat, as well as the surrounding epigenetic environment, including DNA methylation and chromatin structure. Gender-, tissue-, developmental- and locus-specific cis-elements in conjunction with trans-factors may facilitate instability through the processes of DNA replication, repair and/or recombination. Here we review the available human data that supports the involvement of cis-elements in repeat instability with limited reference to model systems. In diverse tissues at different developmental times and at specific loci, repetitive elements display variable levels of instability, suggesting vastly different mechanisms may be responsible for repeat instability amongst the disease loci and between various tissues. PMID- 14526164 TI - Reactivation of silenced genes and transcriptional therapy. AB - The purpose of this review is to discuss the potential role of "transcriptional therapy" to modulate the expression of target genes in order to treat monogenic as well as multifactorial disorders. In vitro and in vivo experiments with DNA demethylating and histone hyperacetylating drugs are currently performed in several laboratories on a variety of genes. In attempting to place these results into perspective, we divided the target genes into four major categories: (1) single genes with a hypermethylated CpG island; (2) single genes without a CpG island; (3) groups of genes silenced by aberrant DNA methylation; and (4) groups of genes silenced by lack of histone acetylation. We discuss the latest advances in the field of chromatin regulation and, in particular, the role of histone methylation and that of RNA interference in gene silencing. We can expect that in the future regulation of transcription will become an effective treatment for several genetic conditions. PMID- 14526166 TI - Rare fragile sites. AB - Rare folate-sensitive fragile sites are the archetypal trinucleotide repeats. Although the CAG repeat in the androgen receptor, associated with spinobulbar muscular atrophy, was the first to be published in 1991, it was the publication in the same year of the molecular basis of fragile X that focused much attention on trinucleotide repeat expansion as a mutational mechanism. A number of rare fragile sites have had their repeat elements characterised since that time. The so-called "folate-sensitive" fragile sites are likely to be all CCG repeat expansions similar to the fragile X. The folate insensitive fragile sites have more complex longer repeat elements. Only two rare fragile sites (FRAXA and FRAXE) are of unequivocal clinical significance in that they are associated with intellectual disability. PMID- 14526165 TI - Transcription defects induced by repeat expansion: fragile X syndrome, FRAXE mental retardation, progressive myoclonus epilepsy type 1, and Friedreich ataxia. AB - Fragile X mental retardation syndrome, FRAXE mental retardation, Progressive myoclonus epilepsy Type I, and Friedreich ataxia are members of a larger group of genetic disorders known as the Repeat Expansion Diseases. Unlike other members of this group, these four disorders all result from a primary defect in the initiation or elongation of transcription. In this review, we discuss current models for the relationship between the expanded repeat and the disease symptoms. PMID- 14526167 TI - The rare human fragile site 16B. AB - The rare human fragile site 16B (FRA16B) has been found to occur spontaneously. Its expression in lymphocyte cultures can also be induced or greatly enhanced by addition of chemicals which are known to bind to AT-rich DNA regions. Following optimal treatment with 150 microg/ml berenil 24 h prior to fixation, the heterozygote frequency of FRA16B is found to be about 5% in populations of European descent. Thus, FRA16B represents the most common of the rare fragile sites. Consistent with cytogenetic observations, the molecular characterization of FRA16B revealed that it is an amplified 33-base pair AT-rich minisatellite repeat. These interindividually variable, extremely large repeat expansions of 15 70 kb in size do not seem to interfere with the expression of genes essential for human development since heterozygotes and homozygotes for FRA16B are normal. PMID- 14526168 TI - Forgotten fragile sites and related phenomena. AB - Fragile sites have been recognised since 1965. Extensive molecular characterisation of them has occurred in the last decade. Yet there are a number of interesting phenomena relating to fragile sites and similar lesions on chromosomes that have not been subject to recent published research. The aim of this short review is to stimulate interest in some of these aspects of fragile sites in the hope that a more complete picture of their nature and properties will emerge. PMID- 14526169 TI - Common fragile sites. AB - Aphidicolin-induced common fragile sites are site-specific gaps or breaks seen on metaphase chromosomes after partial inhibition of DNA synthesis. These fragile sites were first recognized during the early studies of the fragile X syndrome and are induced by the same conditions of folate or thymidylate stress used to induce the fragile X site. Common fragile sites are normally stable in cultured human cells. However, following induction with replication inhibitors, they display a number of characteristics of unstable and highly recombinogenic DNA. From the many studies that have cloned and characterized fragile sites, it is now known that these sites extend over large regions, are associated with genes, exhibit late or delayed replication, and contain regions of high flexibility but are otherwise unremarkable in sequence. Studies showing that fragile sites and their associated genes are frequently deleted or rearranged in cancer cells have clearly demonstrated their importance in genome instability in tumorigenesis. Yet until recently, very little was known about the molecular mechanisms involved in their stability. Recent findings showing that the key checkpoint genes ATR and BRCA1 are critical for genome stability at fragile sites have shed new light on these mechanisms and on the biological significance of common fragile sites. PMID- 14526170 TI - WWOX, the common chromosomal fragile site, FRA16D, cancer gene. AB - Gross chromosomal rearrangements and aneuploidy are among the most common somatic genomic abnormalities that occur during cancer initiation and progression, in particular in human solid tumor carcinogenesis. The loss of large chromosomal regions as consequence of gross rearrangements (e.g. deletions, monosomies, unbalanced translocations and mitotic recombination) have been traditionally associated with the existence of tumor suppressor genes within the areas affected by the loss of genetic material. The long arm of chromosome 16 was identified as being frequently associated with structural abnormalities in multiple neoplasias, that led us to focus attention on the detailed genetic dissection of this region resulting in the cloning of the putative tumor suppressor gene, WWOX (WW domain containing Oxidoreductase). Interestingly, the WWOX gene resides in the very same region as that of the common chromosomal fragile site 16D (FRA16D). The WWOX gene encodes a protein that contains two WW domains, involved in protein-protein interactions, and a short chain dehydrogenase (SDR) domain, possibly involved in sex-steroid metabolism. We have identified the WWOX WW domain ligand as the PPXY motif confirming the biochemical activity of this domain. WWOX normally resides in the Golgi and we will demonstrate that Golgi localization requires an intact SDR. Inactivation of the WWOX gene during tumorigenesis can occur by homozygous deletions and possibly mutation, however, aberrantly spliced forms of WWOX mRNA have been observed even when one allele is still intact. The aberrantly spliced mRNAs have deletions of the exons that encode the SDR and these WWOX protein isoforms display abnormal intracellular localization to the nucleus possibly functioning as dominant negative inhibitors of full length WWOX. Thus, generation of aberrant transcripts of WWOX may represent a novel mechanism to functionally inactivate WWOX without genomic alteration of the remaining allele. In this article we will review the cloning and identification of WWOX as the target of FRA16D. In addition, we will discuss the possible biochemical functions of WWOX and present evidence that ectopic WWOX expression inhibits tumor growth. PMID- 14526172 TI - Expression of the FMR1 gene. AB - Fragile X syndrome is a trinucleotide repeat disorder in which a (CGG)n element located within the 5' untranslated region of the Fragile X Mental Retardation 1 (FMR1) gene expands to more than 200 copies (full mutation) and becomes hypermethylated. Such expansions are accompanied by the failure to produce FMR1 protein (FMRP), resulting in the fragile X phenotype. For smaller (premutation) expansions (approximately 55-200 repeats), FMR1 mRNA and FMRP levels had been assumed to be normal; however, our group and others have recently demonstrated that FMR1 mRNA levels are elevated in cells harboring premutation alleles. Moreover, mRNA levels remain elevated in fragile X males with partially methylated full mutation alleles. Finally, some fragile X males with hypermethylated, full mutation alleles continue to produce FMR1 mRNA, despite the expectation that those genes should be silent. These observations all point to a complex mechanism of expression of the FMR1 gene, one that provides a more consistent foundation for the spectrum of clinical involvement. An FMRP deficit is observed in all categories of fragile X individuals, including carriers of the premutation and partially methylated full mutation alleles. These results demonstrate that lowered FMRP levels, in the absence of methylation-coupled silencing of the FMR1 gene, are not caused by reduced transcriptional activity, but rather by a reduced efficiency of translation. PMID- 14526171 TI - Understanding fragile X syndrome: insights from animal models. AB - The fragile X mental retardation syndrome is caused by large methylated expansions of a CGG repeat in the FMR1 gene leading to the loss of expression of FMRP, an RNA-binding protein. FMRP is proposed to act as a regulator of mRNA transport or translation that plays a role in synaptic maturation and function. To study the physiological function of the FMR1 protein, mouse and Drosophila models have been developed. The loss-of-function mouse model shows slightly enlarged testes, a subtle behavioral phenotype, and discrete anomalies of dendrite spines similar to those observed in brains of patients. Studies in Drosophila indicate that FXMR plays an important role in synaptogenesis and axonal arborization, which may underlie the observed deficits in flight ability and circadian behavior of FXR mutant flies. The relevance of these studies to our understanding of fragile X syndrome is discussed. PMID- 14526173 TI - FMR2 function: insight from a mouse knockout model. AB - The FMR2 gene is dysregulated by the fragile X E triplet repeat expansion in patients with FRAXE mental retardation syndrome. A CCG triplet, located in the 5' untranslated region of the FRAXE gene undergoes expansion and methylation in these patients, eliminating detectable gene transcription. FRAXE syndrome is distinct from fragile X syndrome, a more common genetic form of mental retardation caused by expansion and methylation of a similar repeat in the FMR1 gene located 600 kb proximal to FRAXE. FRAXE syndrome is rare, and patients' phenotypes are highly variable, leading to difficulties with predicting specific FMR2 functions based on the human disease. Recently, Lilliputian(Lilli), a Drosophila FMR2 orthologue, was identified; this gene has been linked with several signal transduction pathways, including the transforming growth factor beta (TGF-beta) pathway, the Raf/MEK/MAP kinase (MAPK) pathway, and the P13K/PKB pathway. Mutation of Lilli shows defects in germinal band extension, cytoskeletal structure, cell growth, and organ development. The Lilli gene suggests possible functions for FMR2 (and related genes) in humans and mice, but cannot predict specific functions. Modeling FMR2 mutation in the mouse will be useful to understand specific functions of this gene in vertebrates. This review presents what has been learned thus far from the FMR2 knockout mouse model and suggests future studies on this model in order to compare it with the human FRAXE mental retardation disorder, Lilli mutants in Drosophila and other mouse models of genes in this family. PMID- 14526174 TI - Instability of the fragile X syndrome repeat in mice: the effect of age, diet and mutations in genes that affect DNA replication, recombination and repair proficiency. AB - Repeat expansion diseases such as fragile X syndrome (FXS) result from increases in the size of a specific tandem repeat array. In addition to large expansions, small changes in repeat number and deletions are frequently seen in FXS pedigrees. No mouse model accurately recapitulates all aspects of this instability, particularly the occurrence of large expansions. This may be due to differences between mice and humans in CIS and/or TRANS-acting factors that affect repeat stability. The identification of such factors may help reveal the expansion mechanism and allow the development of suitable animal models for these disorders. We have examined the effect of age, dietary folate, and mutations in the Werner's syndrome helicase (WRN) and TRP53 genes on FXS repeat instability in mice. WRN facilitates replication of the FXS repeat and enhances Okazaki fragment processing, thereby reducing the incidence of processes that have been suggested to lead to expansion. p53 is a protein involved in DNA damage surveillance and repair. We find two types of repeat instability in these mice, small changes in repeat number that are seen at frequencies approaching 100%, and large deletions which occur at a frequency of about 10%. The frequency of these events was independent of WRN, p53, parental age, or folate levels. The large deletions occur at the same frequency in mice homozygous and heterozygous for the repeat suggesting that they are not the result of an interallelic recombination event. In addition, no evidence of large expansions was seen. Our data thus show that the absence of repeat expansions in mice is not due to a more efficient WRN protein or p53-mediated error correction mechanism, and suggest that these proteins, or the pathways in which they are active, may not be involved in expansion in humans either. Moreover, the fact that contractions occur in the absence of expansions suggests that these processes occur by different mechanisms. PMID- 14526175 TI - Spinocerebellar ataxia type 6 and episodic ataxia type 2: differences and similarities between two allelic disorders. AB - Spinocerebellar ataxia type 6 (SCA6) is one of three allelic disorders caused by mutations of CACNA1A gene, coding for the pore-forming subunit of calcium channel type P/Q. SCA6 is associated with small expansions of a CAG repeat at the 3' end of the gene, while point mutations are responsible for its two allelic disorders (Episodic Ataxia type 2 and Familial Hemiplegic Migraine). Genetic, clinical, pathological and pathophysiological data of SCA6 patients are reviewed and compared to those of other SCAs with expanded CAG repeats as well as to those of its allelic channelopathies, with particular reference to Episodic Ataxia type 2. Overall SCA6 appears to share features with both types of disorders, and the question as to whether it belongs to polyglutamine disorders or to channelopathies remains unanswered at present. PMID- 14526176 TI - Spinocerebellar ataxia 7 (SCA7). AB - Spinocerebellar ataxia 7 (SCA7) is a progressive autosomal dominant neurodegenerative disorder characterized clinically by cerebellar ataxia associated with progressive macular dystrophy. The disease affects primarily the cerebellum and the retina, but also many other CNS structures as the disease progresses. SCA7 is caused by expansion of an unstable trinucleotide CAG repeat encoding a polyglutamine tract in the corresponding protein, ataxin-7. Normal SCA7 alleles contain 4-35 CAG repeats, whereas pathological alleles contain from 36-306 CAG repeats. SCA7 has a number of features in common with other diseases with polyglutamine expansions: (i) the appearance of clinical symptoms above a threshold number of CAG repeats (>35); (ii) a correlation between the size of the expansion and the rate of progression of the disease: the larger the repeat, the faster the progression; (iii) instability of the repeat sequence (approximately 12 CAG/transmission) that accounts for the marked anticipation of approximately 20 years/generation. The CAG repeat sequence is particularly unstable and de novo mutations can occur during paternal transmissions of intermediate size alleles (28-35 CAG repeats). This can explain the persistence of the disease in spite of the anticipation that should have resulted in its extinction. PMID- 14526177 TI - Insights into the molecular basis of polyglutamine neurodegeneration from studies of a spinocerebellar ataxia type 7 mouse model. AB - Spinocerebellar ataxia type 7 (SCA7) is one member of a growing list of neurodegenerative disorders that are all caused by CAG repeat expansions that produce disease by encoding elongated polyglutamine tracts in a variety of apparently unrelated proteins. In this review, we provide an overview of our efforts to determine the molecular basis of polyglutamine neurotoxicity in SCA7 by modeling this polyglutamine repeat disorder in mice. We discuss how our SCA7 mouse model develops a phenotype that is reminiscent of the retinal and cerebellar disease pathology seen in human patients. All of these findings are considered in the context of numerous other models of polyglutamine disease pathology in mice and other organisms, together with various other in vitro and biochemical studies. We present the competing hypotheses of polyglutamine disease pathogenesis, and explain how our studies of SCA7 brainstem and retinal degeneration using this mouse model have yielded insights into possible mechanisms and pathways of polyglutamine disease pathology. In addition to illustrating how our SCA7 mouse model has allowed us to develop and advance notions of disease pathogenesis, we propose a model of polyglutamine molecular pathology that attempts to integrate the key observations in the field. We close by describing why our SCA7 mouse model should be useful for the next phase of polyglutamine disease research--the development of therapies, and predict that this stage of experimentation will continue to rely heavily on the mouse. PMID- 14526178 TI - Molecular genetics of spinocerebellar ataxia type 8 (SCA8). AB - We previously reported that a transcribed but untranslated CTG expansion causes a novel form of ataxia, spinocerebellar ataxia type 8 (SCA8) (Koob et al., 1999). SCA8 was the first example of a dominant spinocerebellar ataxia that is not caused by the expansion of a CAG repeat translated into a polyglutamine tract. This slowly progressive form of ataxia is characterized by dramatic repeat instability and a high degree of reduced penetrance. The clinical and genetic features of the disease are discussed below. PMID- 14526179 TI - SCA10 and ATTCT repeat expansion: clinical features and molecular aspects. PMID- 14526180 TI - Why is SCA12 different from other SCAs? AB - Spinocerebellar ataxia type 12 (SCA12), now described in European-American and Asian (Indian) pedigrees, is unique among the SCAs from clinical, pathological, and molecular perspectives. Clinically, the distinguishing feature is early and prominent action tremor with variability in other signs. Pathologically, brain MRIs also suggest variability, with prominent cortical as well as cerebellar atrophy. Genetically, SCA12 is caused by a CAG repeat expansion that does not encode polyglutamine; we speculate that the mutation may affect expression of the gene PPP2R2B, which encodes a brain-specific regulatory subunit of the protein phosphatase PP2A. PMID- 14526181 TI - The hereditary spinocerebellar ataxias in Japan. AB - In Japan, multiple system atrophy (MSA) accounts for 40% of all spinocerebellar ataxias (SCAs) and hereditary disorders account for 30%. Among the latter, autosomal dominant disorders are common and recessive ataxias are rare. Although the frequency of SCA genotypes differs between geographic regions throughout Japan, SCA6, SCA3/MJD, and DRPLA are the three major disorders, while SCA7, SCA8, SCA10, SCA12, and SCA17 are infrequent or almost undetected. SCA1 predominantly occurs in the northern part of Japan. Overall, 20-40% of dominant SCAs are due to unknown mutations. From this cluster, pure cerebellar ataxias linked with the SCA4, SCA14, and SCA16 locus have been isolated. Among the recessive SCAs, patients with AVED and EAOH have been detected. However, FRDA associated with GAA repeat expansion in the frataxin gene has not been reported so far. PMID- 14526182 TI - A cerebellar tremor/ataxia syndrome among fragile X premutation carriers. AB - Fragile X syndrome is a neurodevelopmental disorder that is not known to have any progressive neurological sequelae in adulthood. However, a neurological condition involving intention tremor, ataxia, and cognitive decline has recently been identified among older male carriers of premutation alleles of the FMR1 gene. This condition is clinically distinct from fragile X syndrome and arises through a different molecular mechanism involving the same gene (FMR1). Characteristic findings on magnetic resonance imaging include cerebral and cerebellar volume loss and altered signal intensities of the middle cerebellar peduncles. A striking feature of this fragile X-associated tremor/ataxia syndrome is the presence of ubiquitin-positive neuronal and astroglial intranuclear inclusions. Unlike the CAG repeat expansion diseases, which lead to altered protein products, there is no known protein abnormality among FMR1 premutation carriers. Thus, inclusion formation may reflect a gain-of-function effect of the FMR1 mRNA or the CGG repeat itself. Finally, since this syndrome may represent one of the more common single-gene causes of tremor, ataxia, and dementia among older males, FMR1 DNA testing should be considered when evaluating adult patients with tremor/ataxia. PMID- 14526183 TI - The epilepsy, the protease inhibitor and the dodecamer: progressive myoclonus epilepsy, cystatin b and a 12-mer repeat expansion. AB - Progressive myoclonus epilepsy 1 (EPM1) or Unverricht-Lundborg disease is a human autosomal recessive neurodegenerative disorder caused by mutations in cystatin B (CSTB). The CSTB gene maps to human chromosome 21 and encodes an inhibitor of lysosomal cysteine proteases. Five point mutations have been found, two of which are seen in numerous unrelated patients. However, the main CSTB mutation in EPM1, even among patients of different ethnic origins, is an expansion of a dodecamer repeat (CCCCGCCCCGCG) in the 5' flanking area of CSTB. Most normal alleles contain either two or three repeats, while rarer normal alleles that are highly unstable contain between 12 and 17 repeats. Mutant expanded alleles have been reported to contain between 30 and 80 copies and are also highly unstable, particularly via parental transmission. There is no apparent correlation between mutant repeat length and disease phenotype. While the repeat expansion is outside the CSTB transcriptional unit, it results in a marked decrease in CSTB expression, at least in certain cell types in vitro. CSTB homozygous knockout mice show some parallels to the phenotype of human EPM1 including myoclonic seizures, development of ataxia and neuropathological changes associated with cell loss via apoptosis. Loss of CSTB function due to mutations is consistent with the observed neurodegenerative pathology and phenotype, but the functional link to the epileptic phenotype of EPM1 remains largely unknown. PMID- 14526184 TI - Spontaneous chromosome loss and colcemid resistance in lymphocytes from patients with myotonic dystrophy type 1. AB - Myotonic Dystrophy type 1 (DM1) is one of the many inherited human diseases whose molecular defect is the expansion of a trinucleotide DNA sequence. DM1 shares with fragile X syndrome (FMR1), another "unstable triplet syndrome", several molecular features not present in the remaining triplet diseases. As FMR1 is also characterised by chromosome instability at the site of the expanded triplet, lymphocytes from DM1 patients and healthy donors were cultured for micronucleus (MN) analysis, in order to verify if DM1 is also prone to chromosome instability. A FISH analysis was also carried out to detect the presence of centromeric sequences in the observed MN. The data indicate that DM1 patients present a percentage of centromere-positive MN significantly higher than controls, suggesting that chromosome loss is the main mechanism underlying the origin of the increased spontaneous instability. To further assess the proneness to instability of cells of DM1 patients, cultures from patients and controls were treated in vitro with growing concentrations of two different mutagens: colcemid, a "pure" aneugen compound whose target is tubulin, and mytomicin C, a strong clastogen. The results show that the patient group is significantly less sensitive to colcemid. These data, together with FISH analysis, suggest the presence, in DM1 patients, of an already damaged tubulin, which becomes no more sensitive to the effect of colcemid and which could be the main defect underlying the aneugenic effects in DM1. PMID- 14526185 TI - Transgenic mouse models for myotonic dystrophy type 1 (DM1). AB - The study of animal models for myotonic dystrophy type 1 (DM1) has helped us to 'de- and reconstruct' our ideas on how the highly variable multisystemic constellation of disease features can be caused by only one type of event, i.e., the expansion of a perfect (CTG)(n) repeat in the DM1 locus on 19q. Evidence is now accumulating that cell type, cell state and species dependent activities of the DNA replication/repair/recombination machinery contribute to the intergenerational and somatic behavior of the (CTG)(n) repeat at the DNA level. At the RNA level, a gain-of-function mechanism, with dominant toxic effects of (CUG)(n) repeat containing transcripts, probably has a central role in DM1 pathology. Parallel study of DM2, a closely related form of myotonic dystrophy, has revealed a similar mechanism, but also made clear that part of the attention should remain focused on a possible role for candidate loss-of-function genes from the DM1 locus itself (like DMWD, DMPK and SIX5) or elsewhere in the genome, to find explanations for clinical aspects that are unique to DM1. This review will focus on new insight regarding structure-function features of candidate genes involved in DM1 pathobiology, and on the mechanisms of expansion and disease pathology that have now partly been disclosed with the help of transgenic animal models. PMID- 14526186 TI - Transgenic mouse models of spinal and bulbar muscular atrophy (SBMA). AB - Spinal and bulbar muscular atrophy (SBMA) is a late-onset motor neuron disease characterized by proximal muscle atrophy, weakness, contraction fasciculations, and bulbar involvement. Only males develop symptoms, while female carriers usually are asymptomatic. A specific treatment for SBMA has not been established. The molecular basis of SBMA is the expansion of a trinucleotide CAG repeat, which encodes the polyglutamine (polyQ) tract, in the first exon of the androgen receptor (AR) gene. The pathologic hallmark is nuclear inclusions (NIs) containing the mutant and truncated AR with expanded polyQ in the residual motor neurons in the brainstem and spinal cord as well as in some other visceral organs. Several transgenic (Tg) mouse models have been created for studying the pathogenesis of SBMA. The Tg mouse model carrying pure 239 CAGs under human AR promoter and another model carrying truncated AR with expanded CAGs show motor impairment and nuclear NIs in spinal motor neurons. Interestingly, Tg mice carrying full-length human AR with expanded polyQ demonstrate progressive motor impairment and neurogenic pathology as well as sexual difference of phenotypes. These models recapitulate the phenotypic expression observed in SBMA. The ligand dependent nuclear localization of the mutant AR is found to be involved in the disease mechanism, and hormonal therapy is suggested to be a therapeutic approach applicable to SBMA. PMID- 14526187 TI - Oculopharyngeal muscular dystrophy: a late-onset polyalanine disease. AB - Oculopharyngeal muscular dystrophy (OPMD) is a muscle disease of late onset associated with progressive ptosis of the eyelids, dysphagia, and unique tubulofilamentous intranuclear inclusions (INIs). OPMD is usually transmitted as an autosomal dominant trait (OMIM 164300). A rarer allelic autosomal recessive form has also been observed (OMIM 257950). Both forms are caused by short (GCG)8 13 expansions in the polyadenylate-binding protein nuclear 1 gene (PABPN1) located on chromosome 14q11.1. The mutations cause the lengthening of an N terminal polyalanine domain. Both slippage and unequal recombination have been proposed as the mutation mechanisms. The size of the mutation has not yet been conclusively shown to inversely correlate with the severity of the phenotype. Mutated PABPN1 proteins have been shown to be constituents of the INIs. The INIs also contain ubiquitin, proteasome subunits, HSP 40, HSP 70, SKIP, and abundant poly(A)-mRNA. The exact mechanism responsible for polyalanine toxicity in OPMD is unknown. Various intranuclear inclusion dependent and independent mechanisms have been proposed based on the major known function of PABPN1 in polyadenylation of mRNA and its shuttling from the nucleus to the cytoplasm. OPMD is one of the few triplet-repeat diseases for which the function of the mutated gene is known. Because of the increasing number of diseases caused by polyalanine expansions and the pathological overlap with CAG/polyglutamine diseases, what pathological insight is gained by the study of OPMD could lead to a better understanding of a much larger group of developmental and degenerative diseases. PMID- 14526188 TI - Molecular analyses of Machado-Joseph disease. PMID- 14526189 TI - The use of transgenic and knock-in mice to study Huntington's disease. AB - The trinucleotide repeat disorders comprise an ever expanding list of diseases, all of which are caused by an unstable expanded trinucleotide repeat tract. Huntington's disease (HD) is a member of this family of diseases and more specifically, is a Type II trinucleotide repeat disorder. This means that the mutation in HD is an unstable expanded polyglutamine repeat tract, which is expressed at protein level. There is no cure or beneficial treatment for this fatal neurodegenerative disorder, and patients suffer from progressive motor, cognitive and psychiatric dysfunction. Recent years has seen the development of many genetic models of HD, which allow study of the early phases of disease process, at several different levels of cell function. In addition, these models are being used to investigate the potential of a variety of therapeutic agents for clinical use. Here we review these findings, and their implication for HD pathogenesis. PMID- 14526190 TI - Mechanisms of neuronal cell death in Huntington's disease. AB - Huntington's disease (HD) is a genetically dominant neurodegenerative condition caused by an unique mutation in the disease gene huntingtin. Although the Huntington protein (Htt) is ubiquitously expressed, expansion of the polyglutamine tract in Htt leads to the progressive loss of specific neuronal subpopulations in HD brains. In this article, we will summarize the current understanding on mechanisms of how mutant Htt can elicit cytotoxicity, as well as how the selective sets of neuronal cell death occur in HD brains. PMID- 14526191 TI - Long-term flexion-extension range of motion of the prodisc total disc replacement. AB - The rationale for total disc replacement is avoidance of the junctional degeneration seen after arthrodesis by preservation of segmental motion. To justify the use of disc prostheses, it is essential to document maintained range of motion (ROM) and sagittal alignment at long-term follow-up. This is a retrospective radiographic study of 42 patients who had placement of 58 first generation Prodisc prostheses at a mean follow-up of 8.7 years. Flexion-extension ROM was measured by Cobb's method. Junctional levels were evaluated for junctional degeneration. Pre- and postoperative global and segmental lordosis were measured. Prognostic patient factors predicting ROM of <2 degrees were evaluated. We observed ROM of at least 2 degrees in 66% of Prodisc prostheses at 8.7-year follow-up, although ROM was less than that reported in asymptomatic normal individuals. Mean ROM for disc prostheses with motion was 7.5 degrees at L3-L4, 6.2 degrees at L4-L5, and 4.1 degrees at L5-S1. Mean ROM for all prostheses was 3.8 degrees. The incidence of radiographic junctional degeneration was 24%, although no patients required surgery for symptomatic junctional degeneration. Mean ROM of prostheses below a degenerated junctional disc was 1.6 degrees compared with 4.7 degrees below a normal junctional disc (P < 0.035). Females were 3.5 times more likely to have ROM of <2 degrees. This is the longest published follow-up study of a lumbar disc replacement. The data show that ROM is preserved at long-term follow-up in the majority of patients. Global and segmental sagittal alignment improve after surgery. Furthermore, there is an association between ROM of disc prostheses and the development of junctional degeneration. PMID- 14526192 TI - Internal stress distribution in cervical intervertebral discs: the influence of an artificial cervical joint and simulated anterior interbody fusion. AB - There is concern that cervical interbody fusion can result in accelerated degenerative changes occurring at adjacent spinal levels. The cervical spine clearly evolved to be mobile. It would seem to be desirable for spinal surgeons to have an alternative to fusion, and spinal arthroplasty is an appealing concept. The Bristol Disc is a mechanical device comprising two articulating components that result in motion with 6 df. It has been shown to have favorable kinematics when compared with intact and fused cadaveric spines. The current study attempts to record changes in the distribution of stresses within cervical intervertebral discs adjacent to the artificial disc or a simulated fusion. The technique used to measure intradiscal stress distributions is based on earlier work by McNally and Adams on lumbar intervertebral discs. The study generated stress profiles through cervical intervertebral discs statically loaded in four different postures in addition to recording changes in intradiscal pressure within both the nucleus and the annulus during flexion. Similar stress profiles were recorded from intact specimens and those with the artificial joint inserted. The artificial joint resulted in reduced stresses in the annulus compared with spines with a simulated fusion. The study demonstrates how different testing conditions can result in researchers being confronted with paradoxical data, and the simulation of muscle forces is recommended. PMID- 14526193 TI - Prospective study evaluating total disc replacement: preliminary results. AB - For many years, there has been an interest in disc replacements. There are a few retrospective studies from Europe reporting promising results for these devices. The purpose of this study is to review the preliminary results of the SB Charite disc replacement device in a prospective study. The study group consists of a consecutive series of our first 57 patients who received the Link SB Charite device implanted through a mini-open anterior approach. The primary clinical indications for disc replacement were single-level symptomatic disc disease at L4 L5 or L5-S1 and no significant facet joint changes. The outcome data include the Visual Analog Scale (VAS) assessing pain intensity and the Oswestry Low Back Pain Disability Questionnaire. Data were collected prior to surgery and at 6 weeks, 3 months, 6 months, and 12 months after surgery. Twenty-four-month follow-up data are currently being collected. The mean VAS score improved 50.8% by the 6-week follow-up, and significant improvement was maintained throughout follow-up. The mean Oswestry score improved 38.7% at the 6-week follow-up, and further slight improvement was noted during the later follow-up periods. This study found that significant improvements were achieved by the 6-week postoperative visit in the pain and disability scores following disc replacement surgery. Significant improvement was maintained through the 12-month follow-up visit. Although these preliminary results are encouraging, long-term follow-up and a greater number of patients are needed to further evaluate the outcome of these devices. PMID- 14526194 TI - Posterior lumbar interbody fusion for degenerative disc disease using a minimally invasive B-twin expandable spinal spacer: a multicenter study. AB - Acquired degenerative disc disease causes gradual disc space collapse, concurrent discogenic or facet-induced pain, and possible compression radiculopathy. Surgical treatment aims to re-expand the intervertebral space and stabilize the involved segment in balanced alignment until fusion is complete. The prevailing methods make use of a twin cage device of predetermined size. Their implantation requires extensive exposure, entailing the sacrifice of posterior stabilizing structures. The procedure also results in significant traction on the dural sac and the cauda equina and is thereby a potential source of neurologic damage. The new expandable spinal spacer (ESS) was designed to mitigate all the shortcomings alluded to above. A prospective multicenter clinical study was conducted of 87 patients with chronic low back pain due to degenerative disc disease, treated by posterior lumbar interbody fusion (PLIF) using a newly designed ESS. The study protocol was approved by the ethics committees of all the participating institutions. The objective was to test the safety and efficacy of the device. Each participant was followed periodically for >1 postoperative year. The ongoing record included intraoperative difficulties and complications, if any, radiologic evidence of fusion and clinical outcome as scored by pre- and postoperative questionnaires pertaining to pain intensity and degree of disability. No dural lacerations or neurologic deficit occurred. There were no mechanical failures of the spacer. Radiologic study demonstrated fusion in all but one patient. Disc space height that averaged 7.53 +/- 2.42 mm before surgery increased to 10.03 +/- 2.00 mm at the time of surgery and stabilized at 9.47 +/- 2.10 mm upon final follow-up. Visual Analog Scale and Oswestry Index decreased by 60% and 58%, respectively. PLIF using the ESS achieves the same ultimate outcome as do other methods currently in use but does not share the handicaps and hazards and is more user-friendly to the surgeon. PMID- 14526195 TI - Comparison of motion restriction and trunk stiffness provided by three thoracolumbosacral orthoses (TLSOs). AB - The amounts of thoracic and lumbar spine motion restriction and passive trunk stiffness provided by three thoracolumbosacral orthoses (TLSOs) (Aspen TLSO, Boston Body Jacket, and CAMP TLSO) were compared. Ten subjects executed maximum trunk flexion, extension, and lateral bending motions. The spine motion was measured noninvasively with a thin strain gauge device (Flexducer), and passive trunk stiffness around the neutral posture was estimated from an electromyography assisted biomechanical model. No significant differences in either the restriction of motion or the amount of added passive trunk stiffness were found between the three orthoses. The subjects also did not perceive any difference in the restriction of motion but rated the Aspen TLSO significantly more comfortable than the other two orthoses. The rigid custom orthosis design may not be important for restricting the spine motion and providing passive trunk stiffness, or there may be other measures that reflect better the function of orthoses. PMID- 14526196 TI - Radiographic analysis of transforaminal lumbar interbody fusion for the treatment of adult isthmic spondylolisthesis. AB - The radiographs of 35 consecutive adult patients with isthmic spondylolisthesis who underwent a transforaminal lumbar interbody fusion (TLIF) with one or two Brantigan carbon fiber cages and pedicle screw instrumentation were evaluated. Anterolisthesis, disk space height, and slip angle were measured in preoperative and postoperative standing neutral radiographs. Anterolisthesis was reduced and disk space height was increased with the TLIF procedure. Average slip angle, however, was not significantly altered. The restoration of lordosis across the listhetic disk space correlated with a more anterior placement of the interbody cage within the disk space. The TLIF technique, performed with the Brantigan cage and pedicle screw instrumentation, appears to be able to restore disk height and reduce forward translation in patients with isthmic spondylolisthesis, but improvement in sagittal alignment is dependent upon anterior placement of the interbody device. PMID- 14526197 TI - The efficacy of anterior spine exposure by an orthopedic surgeon. AB - This retrospective study was designed to document the incidence and types of perioperative complications that occurred with anterior spinal fusion surgery performed solely by an orthopedic spine surgeon. This study is contrasted to previous studies that document complications from anterior approaches performed by an orthopedic surgeon with the assistance of a general or a vascular surgeon. Specifically, the procedures included thoracotomies, thoracolumbar retroperitoneal, and lumbosacral approaches. Our sample consisted of 450 patients who underwent anterior spinal fusion between levels T1 and S1, from 1985 to 1997. Patient and surgery characteristics included age, sex, diagnosis, levels of fusion, blood loss, operative time, hospitalization time, complications, American Society of Anesthesiologists state, assessment of risk factors, previous surgery, and surgical approach used. Average follow-up was 41.69 months, with a minimum of 12 months and a maximum of 132 months. Our results indicated that anterior procedures performed solely by our senior orthopedic surgeon had a lower incidence of complications, less blood loss, and shorter operative time than anterior procedures performed by an orthopedic surgeon and a vascular or a general surgeon. Our findings suggest that the anterior spinal exposure is a safe approach that may be performed solely by a spinal surgeon who is knowledgeable and experienced. PMID- 14526198 TI - Electrophysiologic evaluation of lumbosacral single nerve roots using compound muscle action potentials. AB - Transcutaneous electrical stimulation applied to the vertebral column produces compound muscle action potentials (CMAPs) from the leg muscles. Using this method, we evaluated the efferent pathways of the lumbosacral nerve roots. The subjects were 26 healthy volunteers and 31 patients with lumbar disc herniation (LDH). CMAP recordings were obtained from the bilateral vastus medialis, tibialis anterior, extensor digitorum brevis, and abductor hallucis muscles using low output-impedance stimulation. In normal subjects, the CMAP latency increased linearly with the distance between the stimulating electrode and the recording electrode, with little difference in latency between the left and the right sides in each subject. The CMAP amplitude was significantly lower in the patients with LDH, and the latency was also prolonged when the stimulating electrode was placed above the lesion. This technique may thus be a useful noninvasive method for assessing lumbosacral nerve root function in patients with LDH. PMID- 14526199 TI - Anterior fusion and posterior correction of severe cervical kyphosis using pedicle screw fixation in a patient with neurofibromatosis: a case report. AB - We present a 15-year-old boy with severe cervical kyphosis due to neurofibromatosis (NF), who underwent one-stage anterior fusion and posterior correction using pedicle screw fixation. The kyphosis was corrected from 72 degrees to 35 degrees. At the follow-up 16 months postoperatively, anterior fusion and posterior stabilization had been obtained without correction loss. Cervical pedicle screw fixation is a useful method for the correction of severe cervical kyphosis in patients with NF. PMID- 14526200 TI - Implications of T-cell P-glycoprotein activity during HIV-1 infection and its therapy. AB - OBJECTIVES: P-glycoprotein (P-gp) may reduce antiretroviral efficacy by decreasing disposition of HIV-1 protease inhibitors into tissues and cells. In contrast, P-gp overexpression in vitro can inhibit HIV-1 replication, and some drugs induce P-gp expression. To explore which of these mechanisms predominate in vivo, this study characterized relationships between T-cell P-gp activity and clinical parameters in HIV-infected adults. METHODS: P-gp activity was quantified in total and naive CD4+ and CD8+ T cells of HIV-infected adults by flow cytometry using the substrate dye DiOC2(3). Demographic, virologic, immunologic, and treatment factors were obtained from medical records. Factors associated with P gp activity were identified using multivariate linear regression. RESULTS: A total of 185 subjects (22% female; 34% African American) were studied, of whom 131 (71%) were receiving antiretroviral treatment. There was marked interindividual variability in P-gp activity. By multivariate analysis, higher CD4+ T-cell P-gp activity was associated with lower log10 HIV-1 RNA (P = 0.005), but not treatment or demographic factors. P-gp activity was correlated across T cell subsets. CONCLUSIONS: The inverse relationship between P-gp activity and plasma HIV-1 RNA is most consistent with an inhibitory effect on viral replication rather than drug disposition. Antiretroviral drug class did not independently predict P-gp activity. PMID- 14526201 TI - High frequency of alternative splicing of human genes participating in the HIV-1 life cycle: a model using TSG101, betaTrCP, PPIA, INI1, NAF1, and PML. AB - Alternative splicing may generate splice forms with different biologic roles or missing protein domains implicated in the interaction with HIV-1. To address this issue, 6 human genes were investigated-tumor suppressor gene 101 (TSG101), beta transducin repeats containing protein (betaTrCP), peptidyl-proly cis-trans isomerase, cyclophilin A (PPIA), integrase interactor 1 protein (INI1), Nef associated factor 1 (NAF1), and promyelacytic leukemia (PML)-involved in the viral life cycle and HIV-1 pathogenesis. All 6 genes presented alternative splicing, and a combined bioinformatic and reverse transcription polymerase chain reaction (RT-PCR) analysis identified 27 new variants for a total of 53 splice forms (an average of 9 variants per gene). The predicted frequency of the various splice forms based on expressed sequence tags (EST) analysis corresponded to the semiquantitative findings on RT-PCR analysis for the cell culture systems and for native CD4 cells investigated. Interindividual variation in the frequencies of various splice forms in CD4 T cells from blood donors was observed for INI1. Cell type-specific variation of splice pattern was observed for NAF1. Eight splice forms lacked or modified motifs implicated in the interaction with HIV-1, underscoring the potential interest of assessing alternative splicing when investigating viral cell biology and pathogenesis. PMID- 14526202 TI - Pharmacokinetics of indinavir/ritonavir (800/100 mg) in combination with efavirenz (600 mg) in HIV-1-infected subjects. AB - BACKGROUND: Addition of efavirenz (600 mg) to indinavir/ritonavir (800/100 mg) results in significant decreases in indinavir levels in healthy volunteers. This study evaluated the steady-state pharmacokinetics of indinavir/ritonavir at 800/100 mg twice daily (bid) in combination with efavirenz at 600 mg once daily (qd) in HIV-infected Thai subjects who used this nucleoside-sparing combination in The HIV Netherlands Australia Thailand Research Collaboration 009 study. METHODS: At week 4 of the study, 12-hour pharmacokinetic profiles for indinavir/ritonavir were obtained for 20 HIV-infected subjects. For efavirenz, the concentrations at 12 hours and 24 hours (Cmin) after dosing were assessed. RESULTS: All subjects (10 males and 10 females) completed the study. The geometric mean area under the concentration versus time curve, Cmin, and maximum plasma concentration of indinavir were 45.7 mg/(L. h) (95% confidence interval [CI], 39.8-52.5), 0.32 mg/L (95% CI, 0.24-0.44), and 11.1 mg/L (95% CI, 9.4 13.0), respectively. A >10-fold variation in indinavir Cmin was observed. All subjects had an indinavir Cmin that was at least comparable with the reported mean population Cmin of indinavir at 800 mg thrice daily without ritonavir (0.15 mg/L). The geometric mean concentration at 12 hours and Cmin of efavirenz were 3.1 mg/L (95% CI, 2.5-3.7) and 2.1 mg/L (95% CI, 1.6-2.6), respectively. CONCLUSIONS: Despite the known pharmacokinetic interaction between efavirenz and indinavir/ritonavir, the combination of indinavir/ritonavir at 800/100 mg bid and efavirenz at 600 mg qd results in adequate minimum concentrations of both indinavir and efavirenz for treatment-naive patients. PMID- 14526203 TI - Persistently biased T-cell receptor repertoires in HIV-1-infected combination antiretroviral therapy-treated patients despite sustained suppression of viral replication. AB - In most HIV-1-infected patients, highly active antiretroviral therapy (HAART) reduces plasma viral load to <50 copies/mL and increases CD4+ T-cell number and function. However, it is still unclear whether alterations of T-cell receptor (TCR) beta-chain variable region (BV) repertoire, tightly related to disease progression, can be fully recovered by long-term treatment with HAART. This study analyzed the evolution of both T-cell subset composition and TCRBV perturbations in chronically HIV-1-infected patients with moderate immunodeficiency during 36 months of HAART. Despite persistently suppressed HIV replication, the rate of CD4+ T-cell repopulation, after an initial burst, progressively declined throughout the study period, resulting in a mean CD4+ T-cell count at the end of follow-up that was still significantly lower in HIV patients than in HIV seronegative controls. This was seen in association with an incomplete restitution of both CD4 and CD8 TCRBV repertoire disruptions and was also demonstrated by the appearance of new TCRBV oligoclonal expansions occurring during HAART. In conclusion, these data indicate that 3 years of fully suppressive HAART may be not adequate to normalize CD4 counts and TCRBV repertoires in patients starting HAART with moderately advanced disease. PMID- 14526204 TI - Neurocognitive function in HIV-infected patients with low weight and weight loss. AB - OBJECTIVE: To determine neurocognitive function in HIV-infected subjects with low weight and weight loss. DESIGN: Cross-sectional cohort. METHODS: Baseline data from male and female participants in longitudinal treatment studies of AIDS wasting were analyzed. Fifty-seven HIV-infected women (age: 38 +/- 5 years, education level: 12.3 +/- 2.3 years) and 24 HIV-infected men (age: 37 +/- 5 years, education level: 13.5 +/- 2.9 years) with weight <90% IBW or loss of >10% of preillness weight maximum underwent standardized neurocognitive testing to measure intellectual functioning, attention, memory, and fine motor dexterity. The z scores were determined using age- and gender-specific normative data. RESULTS: Among women, IQ (87 +/- 13, z score: -0.8 +/- 0.8 SD) and executive functioning scores (-0.7 +/- 1.2 SD) were below average but within normal limits. In contrast, performance (z scores) on measures of verbal learning (-2.5 +/- 1.5 SD), visuospatial abilities (-2.5 +/- 2.0 SD), and motor coordination (-2.3 +/- 3.3 SD) fell significantly below normal limits. Among men, IQ score was 96 +/- 13 SD (z score: -0.3 +/- 0.8 SD). Performance was below average on measures of verbal learning (-1.0 +/- 1.7 SD) and visual construction (-1.6 +/- 1.7 SD) but within normal limits on executive functioning (-0.1 +/- 1.0 SD). CONCLUSIONS: HIV infected women at low weight demonstrate significantly reduced verbal learning, memory, and motor function, whereas HIV-infected men at low weight demonstrate more moderate impairment in verbal learning and other measures of neurocognitive function. PMID- 14526205 TI - Hospitalization rates differ by hepatitis C satus in an urban HIV cohort. AB - OBJECTIVE: To determine whether hepatitis C virus (HCV) infection status affected hospitalization rates, intensive care utilization rates, and discharge diagnoses between 1995 and 2000 in patients with HIV. METHODS: We conducted a prospective cohort study of 3730 HIV patients who were longitudinally followed between 1995 and 2000. All hospitalizations of these patients were classified as an opportunistic illness (OI) using the 1993 indicator diagnoses in the case definition of AIDS, complication of injection drug use (IDU) (abscess, cellulitis, osteomyelitis, bacteremia, endocarditis, and poisoning by analgesics), liver-related complication (acute and subacute necrosis of the liver, chronic liver disease and cirrhosis, liver abscess, hepatic coma, portal hypertension, hepatorenal syndrome, hepatocellular carcinoma, and gastrointestinal bleed), or other. Negative binomial regression was used to assess for risk factors for hospitalization. MAIN OUTCOME MEASURES: Inpatient hospitalization and intensive care utilization rates and discharge diagnoses. RESULTS: Nearly half (42.8%) of our cohort was infected with HCV. Between 1995 and 2000, hospitalization rates for HCV-negative patients decreased from 61.9 to 33.9 per 100 patient-years (PY) of follow-up (P = 0.007). Hospitalization rates decreased between 1995 and 1997 for HCV-positive patients from 55.4 to 43.5 per 100 PY but increased between 1997 and 2000 from 43.5 to 62.9 per 100 PY (P = 0.001). When stratified by diagnostic category, IDU-related complications increased from 13.6 to 18.4 admissions per 100 PY and liver-related complications increased from 5.4 to 26.7 admissions per 100 PY between 1995 and 2000 in HCV positive patients (P < 0.001); however, OIs remained relatively unchanged from 1995 to 2000, with 14.6 to 13.0 hospitalizations per 100 PY. In multivariate analysis, HCV infection (incidence rate ratio [IRR] = 1.75, 95% confidence interval [CI]: 1.47, 2.07), female gender (IRR = 1.56, 95% CI: 1.32, 1.85), age <37 years (IRR = 1.19, 95% CI: 1.01, 1.41), African American ethnicity (IRR = 1.30, 95% CI: 1.05, 1.61), and CD4 cell count <50 cells mm3 (IRR = 2.20, 95% CI: 1.72, 2.83) were predictive of hospitalization. CONCLUSIONS: Our data indicate that hospitalization rates decreased significantly between 1995 and 2000 for HCV negative patients but increased significantly for HCV-positive patients. Hospitalization rates for IDU- and liver-related complications increased during this time interval in coinfected patients. In the era of highly active antiretroviral therapy, HIV/HCV-coinfected patients are more likely to suffer from higher hospitalization rates, which will require more health care resources. PMID- 14526206 TI - Impact of an educational program on efficacy and adherence with a twice-daily lamivudine/zidovudine/abacavir regimen in underrepresented HIV-infected patients. AB - A 24-week open-label clinical trial was conducted in 195 HIV-infected adults commonly underrepresented in research (35% female, 71% African American, 21% Hispanic, and 20% injection drug users [IDUs]) to evaluate the effect of an HIV educational program on efficacy and adherence with a simple, compact, twice-daily triple nucleoside regimen containing a lamivudine (150 mg)/zidovudine (300 mg) combination (COM) tablet plus abacavir (ABC), 300 mg. At baseline, the patients' median plasma HIV-1 RNA level was 4.18 log10 copies/mL and the median CD4+ cell count was 379 cells/mm3. Patients were randomized 1:1 to 4 modules of the Tools for Health and Empowerment HIV education intervention plus routine counseling (EI + RC; n = 96) or to routine counseling alone (RC; n = 99). No differences between the EI + RC and RC treatment arms were observed with respect to the proportion of patients achieving plasma HIV-1 RNA levels <40 copies/mL (60% [33/55] vs. 55% [38/69]; P = 0.529) or <400 copies/mL (80% [44/55] vs. 80% [55/69]; P = 0.689) at week 24 (intent-to-treat observed analysis), increase in median CD4 cell count above baseline at week 24 (78.3 vs. 104.8 cells/mm3; P = 0.498), or mean overall adherence rates as measured by the Medication Event Monitoring System (MEMS) (70% vs. 74%). COM + ABC was generally well tolerated, and no association was observed between interruptions in treatment and the development of ABC hypersensitivity (5 suspected cases). In conclusion, in underrepresented patients, the EI used in this study did not affect the efficacy and adherence results with ABC + COM to any greater degree than did RC. PMID- 14526207 TI - Access to antiretroviral treatment, incidence of sustained therapy interruptions, and risk of clinical events according to sex: evidence from the I.Co.N.A. Study. AB - Objectives of the study were to assess the differences between sexes in the likelihood of starting antiretroviral therapy (ART), in rates of sustained discontinuation from highly active antiretroviral therapy (HAART), and in clinical progression. In a multicenter cohort study (I.Co.N.A. Study), 2323 men and 1335 women previously naive to antiretrovirals were enrolled. As of September 2002, 807 women and 1480 men started ART. The median time to starting ART was 28 weeks for women and 17 weeks for men (P = 0.0003 by log-rank test). This difference was no longer significant after adjusting for either HIV RNA (P = 0.21) or CD4 count (P = 0.28) at enrollment. Women tend to start HAART less frequently than mono/dual ART after adjusting for potential confounders (odds ratio = 0.78, 95% confidence interval [CI]: 0.60-1.01; P = 0.06). Women who started HAART were 1.4 times more likely than men (95% CI: 1.00-1.99; P = 0.05) to interrupt at least 1 drug because of toxicity. Twenty-one percent of women and 19% of men interrupted HAART altogether for more than 3 months (P = 0.3). Clinical progression was observed in 53 women (22.6%) and 137 men (23.4%; P = 0.56). Risk of developing a clinical event was found to be no different between women and men (relative hazard = 0.84, 95% CI: 0.56-1.26; P = 0.40). PMID- 14526208 TI - Impact of a patient education program on adherence to HIV medication: a randomized clinical trial. AB - Patients' knowledge of their HIV condition and its treatment, which has been recognized as a factor that influences adherence to antiretroviral therapy, can be improved through educational programs. This prospective, randomized, controlled trial compared an experimental group that participated in an educational program and a control group with standard care. The study evaluated the impact of an educational intervention on adherence to antiretroviral therapy, patients' knowledge, quality of life, and therapeutic response in patients treated with highly active antiretroviral therapy. Three hundred twenty-six patients were analyzed at inclusion. A higher level of adherence was associated with patients who were older, had higher incomes, and did not smoke. CD4 cell count and plasma viral load were correlated with adherence at entry. The educational intervention had an impact on adherence and knowledge in the experimental group at 6 months, which was maintained at 12 and 18 months. A delayed increase in adherence was observed in the control group at 12 months. No significant impact on quality of life was observed over time. The patients' health status improved in 56% of the experimental group subjects and 50% of the control subjects. However, no significant impact was shown on CD4 cell count and plasma viral load. This study shows that an educational intervention improves adherence to antiretroviral regimens and health status and suggests that it should be initiated early in therapy. PMID- 14526209 TI - Changes in HIV-related preventive behavior in the US population: data from national surveys, 1987-2002. AB - This study estimated 1987-2002 trends in preventive behaviors closely linked to HIV from several large health surveys providing the most recently available data. These behaviors include condom use, dual use of condoms with other contraceptive methods, and HIV testing. Condom use increased throughout the period for adolescents, but there is no evidence of overall increased condom use for adults after the mid-1990s. After 2000, adult condom use with primary partners was low even among those at highest risk. Dual use of condoms with other contraceptive methods was reported by a small and increasing percentage of adolescents and adults. By 2001 a high percentage of US adults reported having been tested at least once, and reproductive-age and pregnant women were tested at a greater rate than others. However, 1 in 4 pregnant women had never been tested for HIV. This review indicates that even after considerable increase in preventive behaviors, it is still possible to identify a relatively large segment of the population that is at risk for transmitting or acquiring HIV. Prevention programs serving high-risk populations need to work toward increasing safe sex practices with main partners and HIV testing among the never-tested, particularly reproductive-age women. PMID- 14526210 TI - Condom use for preventing HIV infection/AIDS in sub-Saharan Africa: a comparative multilevel analysis of Uganda and Tanzania. AB - This study explored the relationships between individual-, household-, and community-level variables and condom use to prevent HIV infection in women and men in Uganda and Tanzania using multilevel modeling. Using data from the Demographic and Health Surveys for Uganda (1995) and Tanzania (1996) as well as data collected by the MEASURE Evaluation Project at the Carolina Population Center for Tanzania (1996 and 1999), the study found higher condom use among men than women. There was also heterogeneity in condom use among different clusters for both women and men. More specifically, women and men living in clusters with higher indicators of development were more likely to use condoms to prevent HIV infection. In addition, condom use was much more prevalent in areas where health care services were nearby (0-5 km). In addition, condom use was more common among women (but not men) who lived in clusters where HIV/AIDS testing, counseling, and treatment were provided. The results further revealed that education improved condom use; however, the effect of education was considerably reduced in the models that included HIV/AIDS knowledge and cluster-level variables. The positive effect of household wealth on condom use also diminished after controlling for the effects of the knowledge and cluster-level factors. Knowledge about HIV and perceiving oneself to be at risk for contracting HIV infection improved condom use. PMID- 14526211 TI - AIDS vaccines that allow HIV-1 to infect and escape immunologic control: a mathematic analysis of mass vaccination. AB - Cytotoxic T lymphocyte (CTL)-based HIV vaccine concepts shown to reduce viremia and postpone disease but not to prevent infection in monkeys are currently in human phase 1 trials. To evaluate the potential efficacy of vaccines that cannot prevent HIV-1 to infect and escape immunologic control, we designed a mathematic model that correlates the level of viremia to both infectiousness and disease progression. We speculate that vaccinees will have a virologic set point and disease progression rates comparable to untreated HIV-1-infected individuals with the best prognosis. Our model (illustrated with R0 = 3) shows that a sexually active population can ultimately be reduced to 26% of its initial size as a result of AIDS-related mortality in the absence of treatment or vaccination. Start of vaccination when HIV-1 prevalence is still low might postpone the peak incidence of infection and the dramatic decline in population size by up to 22 years. In conclusion, CTL-based vaccines that do not prevent HIV-1 infection but do postpone the time to onset of AIDS have considerable potential to curb the spread of HIV-1 and to postpone high AIDS-related mortality on a population level. The number of long-term survivors is substantially increased only when vaccination is initiated early in an AIDS epidemic, however. PMID- 14526212 TI - Epidemiologic modeling to evaluate prevention of mother-infant HIV transmission in Ontario. AB - OBJECTIVES: To evaluate the impact of the Ontario HIV screening program to reduce mother-infant HIV transmission, this study estimated the proportion of preventable transmissions that were prevented. METHODS: Using an iterative spreadsheet model, incidences of HIV infection, AIDS, and AIDS mortality in Ontario women were estimated by exposure category. The number of HIV-infected infants born to HIV-infected mothers was then estimated from conception and abortion rates of HIV-infected women of childbearing age and surveillance data. Finally, the proportion of HIV-infected mothers who received antiretroviral prophylaxis (ARP) was assessed. RESULTS: HIV prevalence in 2001 among women of childbearing age was 1.05 per 1000. From 1984-2001, 764 infants were born to HIV infected mothers and 180 were infected. From mid-1994-2001, 214 (39%) of the estimated 544 HIV-infected mothers were diagnosed; almost all received ARP. Of 118 preventable infections among infants born in this period, 39 (33%) were prevented. In 2001, only 46% of preventable infections were prevented and 11 preventable transmissions occurred. CONCLUSIONS: HIV prevalence among women in Ontario increased >4-fold from 1990 to 2001. Fewer than half of HIV-infected mothers received ARP and preventable HIV infections continue to occur. Measures to further increase uptake of prenatal HIV screening must be instituted. PMID- 14526213 TI - Estimated number of children left motherless by AIDS in the United States, 1978 1998. AB - When a mother dies of AIDS, basic needs of her children may be left unmet. To estimate the number and characteristics of maternal AIDS orphans in the United States, demographic techniques were applied to data from several sources. From the national HIV/AIDS surveillance system, reporting delays were adjusted for the number of deaths among women aged 15-44 diagnosed with AIDS through 1998 and reported as deceased by December 1999. No fertility was assumed in the year preceding death. To the adjusted number of deaths the annual age- and race specific cumulative fertility and infant mortality rates from national vital statistics were applied. A perinatal infection rate of 25% was assumed among children born through 1994, and 10% among children born after 1994. Through 1998, 51,473 women died leaving 97,376 children motherless. Of the estimated 76,661 87,0018 uninfected children, 83% were younger than 21 years when orphaned. After increasing each year, the annual number of orphaned children younger than 21 years peaked in 1995. In 1998, between 4252-4489 uninfected youth were added to 47,863-54,025 existing orphans younger than age 21. Due to declines in AIDS deaths, the annual number of children orphaned by AIDS has declined. Nevertheless, each year thousands of youth are orphaned. PMID- 14526215 TI - Lack of severe acute respiratory syndrome in 19 AIDS patients hospitalized together. PMID- 14526214 TI - Growth, morbidity, and mortality in a cohort of institutionalized HIV-1-infected African children. AB - OBJECTIVE: As a result of the HIV epidemic in Africa, much debate exists on whether institutionalized compared with community-based care provides optimum management of infected children. Previous reports calculated 89% mortality by age 3 years among outpatients in Malawi. No similar data are available for infected children in institutionalized care. We characterized patterns of morbidity and mortality among HIV-1-infected children residing at an orphanage in Nairobi. METHODS: Medical records for 174 children followed over 5 years were reviewed. Mortality was analyzed by Kaplan-Meier methods with adjustment to account for survival in the community before admission. Anthropometric indices were calculated to include mean z scores for weight for length and length for age. Low indices reflected wasting and stunting. Opportunistic infections were documented. RESULTS: Of 174 children, 64 had died. Survival was 70% at age 3 years. Morbidity included recurrent respiratory tract infections, gastroenteritis, parotitis, and lymphoid interstitial pneumonitis. No new cases of tuberculosis disease were noted after admission. Mean z scores for length for age suggested overall stunting (z = -1.65). Wasting was not observed (z = -0.39). CONCLUSION: The optimal form of care for HIV-infected children in resource-poor settings may be the development of similar homes. Absence of tuberculosis disease in long standing residents may have contributed to improved survival. Stunting in the absence of wasting implied that growth was compromised by opportunistic infections and other cofactors. PMID- 14526216 TI - Assessment of the relative potency of emtricitabine and lamivudine. PMID- 14526217 TI - Diarrhea related to lopinavir is most often mild and not related to plasma drug levels. PMID- 14526218 TI - Increased incidence of lung neoplasms among HIV-infected men and the need for improved prevention. PMID- 14526219 TI - Short-term lipid changes associated with highly active antiretroviral therapy in naive HIV-infected patients. PMID- 14526220 TI - Health assessment of HIV-infected refugees. PMID- 14526221 TI - Overexpression of HDJ-2 protects astrocytes from ischemia-like injury and reduces redistribution of ubiquitin staining in vitro. AB - HDJ-2, a member of the HSP40 family, functions as a cochaperone to promote protein folding both by binding to unfolded polypeptides and by regulating the activity of HSP70. This study tested whether HDJ-2 overexpression alone could provide significant protection from ischemia-like injury. Primary mouse astrocyte cultures were infected with an HDJ-2 encoding retroviral vector or control retrovirus lacking HDJ-2. Expression of HDJ-2 was confirmed by immunohistochemical staining and immunoblotting. Injury paradigms to mimic ischemia were glucose deprivation (GD) for 24 hours and oxygen-glucose deprivation (OGD) for 8 hours. Cell death was determined by trypan blue exclusion and cell counting. Overexpression of HDJ-2 alone significantly reduced astrocyte injury after both GD and OGD, independent of an elevation in HSP70. To further search for the mechanism of HDJ-2 protection, cultured astrocytes allowed to recover 16 hours after 8 hours GD were labeled with a monoclonal antiubiquitin antibody that recognizes both free ubiquitin and ubiquitinated proteins. The immunolabeling pattern changed from a relatively even distribution in both nuclei and cytoplasm in control cells to heterogeneous aggregates and marked reduction of nuclear staining in most of the cells after GD. When HDJ-2 was overexpressed, the number of cells with evidence of protein aggregation was significantly reduced. Thus, blocking protein aggregation may be an important mechanism by which HDJ-2 protects cells from damage. PMID- 14526222 TI - Oxidative damage to the endoplasmic reticulum is implicated in ischemic neuronal cell death. AB - The endoplasmic reticulum (ER), which plays important roles in apoptosis, is susceptible to oxidative stress. Because reactive oxygen species (ROS) are robustly produced in the ischemic brain, ER damage by ROS may be implicated in ischemic neuronal cell death. We induced global brain ischemia on wild-type and copper/zinc superoxide dismutase (SOD1) transgenic rats and compared ER stress and neuronal damage. Phosphorylated forms of eukaryotic initiation factor 2 alpha (eIF2 alpha) and RNA-dependent protein kinase-like ER eIF2 alpha kinase (PERK), both of which play active roles in apoptosis, were increased in hippocampal CA1 neurons after ischemia but to a lesser degree in the transgenic animals. This finding, together with the finding that the transgenic animals showed decreased neuronal degeneration, indicates that oxidative ER damage is involved in ischemic neuronal cell death. To elucidate the mechanisms of ER damage by ROS, we analyzed glucose-regulated protein 78 (GRP78) binding with PERK and oxidative ER protein modification. The proteins were oxidatively modified and stagnated in the ER lumen, and GRP78 was detached from PERK by ischemia, all of which were attenuated by SOD1 overexpression. We propose that ROS attack and modify ER proteins and elicit ER stress response, which results in neuronal cell death. PMID- 14526223 TI - Monoubiquitination and cellular distribution of XIAP in neurons after traumatic brain injury. AB - XIAP is a member of the inhibitor of apoptosis (IAP) gene family that, in addition to suppressing cell death by inhibition and polyubiquitination of caspases, is involved in an increasing number of signaling cascades. Moreover, the function and regulation of XIAP in the central nervous system (CNS) is poorly understood. In this study, the authors investigated the cell-type expression, the subcellular distribution, ubiquitination of XIAP, and levels of Smac/DIABLO in the normal adult rat brain and in brains subjected to moderate traumatic brain injury (TBI). In the normal brain, XIAP was predominantly expressed in the perinuclear region of neurons. Traumatized brains showed dramatic alterations in cellular and regional expression of XIAP early after injury. Stereologic analyses of the number of XIAP-positive cells within the hippocampus of both hemispheres showed a biphasic response. Immunoprecipitation and immunoblots of extracts derived from different brain regions demonstrated that a single ubiquitin modifies XIAP. Normal cortex contained significantly higher levels of monoubiquitinated XIAP than hippocampus. TBI induced alterations in levels of monoubiquitinated XIAP that correlated with changes in XIAP distribution and immunoreactivity, suggesting that monoubiquitination of XIAP may be a regulator of XIAP location or activity. Similar levels of Smac/DIABLO were present in lysates of normal and traumatized brains. These data demonstrate for the first time a region-specific regulation of XIAP monoubiquitination in the normal adult rat brain, and after TBI, that may be a key event in the regulation of XIAP function contributing to the pathogenesis following injury. PMID- 14526224 TI - Translocation of apoptosis-inducing factor in vulnerable neurons after transient cerebral ischemia and in neuronal cultures after oxygen-glucose deprivation. AB - Loss of mitochondrial membrane integrity and the resulting release of apoptogenic factors may play a critical role in mediating hippocampal neurodegeneration after transient global ischemia. In the present study, the authors have cloned and characterized the rat cDNA encoding apoptosis-inducing factor (AIF), an intramitochondrial protein that promotes cell death in a caspase-independent manner upon release into nonmitochondrial compartments. In contrast to the expression patterns of a number of apoptosis-regulatory gene products during brain development, the expression of AIF protein increases gradually with brain maturation and peaks in adulthood. In a rat model of transient global ischemia, AIF was found to translocate from mitochondria to the nucleus in the hippocampal CA1 neurons after ischemia and to manifest a DNA-degrading activity that mimicked the purified AIF protein and was inhibitable by AIF immunodepletion. The temporal profile of AIF translocation after ischemia (24 to 72 hours) coincided with the induction of large-scale DNA fragmentation at the size of 50 kbp, a well characterized hallmark of AIF-like activity but preceded the formation of internucleosomal DNA fragmentation (72 hours), a DNA degradation associated with the terminal stage of cell death. Further, the nuclear translocation of AIF after ischemia was not blocked by inhibiting caspase-3/-7 activities, but, as shown in neuronal cultures that were challenged with transient oxygen-glucose deprivation, it can be prevented by intracellular delivery of the mitochondria-associated antiapoptotic protein Bcl-xL. The results presented here strongly suggest that mitochondrial release of AIF may be an important factor, in addition to the previously reported cytochrome c and Smac, which could contribute to the selective vulnerability of CA1 neurons to transient global ischemic injury. PMID- 14526225 TI - Reduced cerebral injury in CRH-R1 deficient mice after focal ischemia: a potential link to microglia and atrocytes that express CRH-R1. AB - Corticotropin releasing hormone (CRH) and its family of related peptides are involved in regulating physiologic responses to multiple stressors, including stroke. Although CRH has been implicated in the exacerbation of injury after stroke, the mechanism remains unclear. After ischemia, both excitotoxic damage and inflammation contribute to the pathology of stroke. CRH is known to potentiate excitotoxic damage in the brain and has been shown to modulate inflammatory responses in the periphery. Here the present authors examine the relative contribution of the two known CRH receptors, CRH-R1 and CRH-R2, to ischemic injury using CRH receptor knockout mice. These results implicate CRH-R1 as the primary mediator of ischemic injury in this mouse model of stroke. In addition, the authors examine a potential role for CRH in inflammatory injury after stroke by identifying functional CRH receptors on astrocytes and microglia, which are cells that are known to be involved in brain inflammation. By single cell PCR, the authors show that microglia and astrocytes express mRNA for both CRH-R1 and CRH-R2. However, CRH-R1 is the primary mediator of cAMP accumulation in response to CRH peptides in these cells. The authors suggest that astrocytes and microglia are cellular targets of CRH, which could serve as a link between CRH and inflammatory responses in ischemic injury via CRH-R1. PMID- 14526226 TI - Neuroprotective effects of insulin-like growth factor-binding protein ligand inhibitors in vitro and in vivo. AB - The role of brain insulin-like growth factors (IGFs) and IGF binding proteins (IGFBPs) in neuroprotection was further investigated using in vitro and in vivo models of cerebral ischemia by assessing the effects of IGF-I, IGF-II, and high affinity IGFBP ligand inhibitors (the peptide [Leu24, 59, 60, Ala31]hIGF-I (IGFBP LI) and the small molecule NBI-31772 (1-(3,4-dihydroxybenzoyl)-3-hydroxycarbonyl 6, 7-dihydroxyisoquinoline), which pharmacologically displace and elevate endogenous, bioactive IGFs from IGFBPs. Treatment with IGF-I, IGF-II, or IGFBP-LI (2 microg/mL) significantly (P < 0.05) reduced CA1 damage in organotypic hippocampal cultures resulting from 35 minutes of oxygen and glucose deprivation by 71%, 60%, and 40%, respectively. In the subtemporal middle cerebral artery occlusion (MCAO) model of focal ischemia, intracerebroventricular (icv) administration of IGF-I and IGF-II at the time of artery occlusion reduced ischemic brain damage in a dose-dependent manner, with maximum reductions in total infarct size of 37% (P < 0.01) and 38% (P < 0.01), respectively. In this model of MCAO, i.c.v. administration of NBI-31772 at the time of ischemia onset also dose-dependently reduced infarct size, and the highest dose (100 microg) significantly reduced both total (by 40%, P < 0.01) and cortical (by 43%, P < 0.05) infarct volume. In the intraluminal suture MCAO model, administration of NBI-31772 (50 microg i.c.v.) at the time of artery occlusion reduced both cortical infarct volume (by 40%, P < 0.01) and brain swelling (by 24%, P < 0.05), and it was still effective when treatment was delayed up to 3 hours after the induction of ischemia. These results further define the neuroprotective properties of IGFs and IGFBP ligand inhibitors in experimental models of cerebral ischemia. PMID- 14526227 TI - Reduction of ischemic brain damage by nitrous oxide and xenon. AB - Neuronal death after ischemia-induced brain damage depends largely upon the activation of the N-methyl-D-aspartate (NMDA) excitatory glutamate receptor that is a target for many putative neuroprotective agents. Whereas the NMDA receptors mediate ischemic brain damage, blocking them is deleterious in humans. Here, the authors investigated whether nitrous oxide or xenon, which are gaseous anesthetics with a remarkably safe clinical profile that have been recently demonstrated as effective inhibitors of the NMDA receptor, may reduce the following: (1) ischemia-induced brain damage in vivo, when given after occlusion of the middle cerebral artery (MCAO), a condition needed to make these potentially neuroprotective agents therapeutically valuable; or (2) NMDA-induced Ca2+ influx in cortical cell cultures, a major critical event involved in excitotoxic neuronal death. The authors have shown that both nitrous oxide at 75 vol% and xenon at 50 vol% reduce ischemic neuronal death in the cortex by 70% and further decrease NMDA-induced Ca2+ influx by 30%. In addition, xenon at 50%, but not nitrous oxide at 75 vol%, further decreases ischemic brain damage in the striatum (a subcortical structure that is known to be resistant to neuroprotective interventions). However, at a higher concentration (75 vol%), xenon exhibits potentially neurotoxic effects. The mechanisms of the neuroprotective and potentially neurotoxic effects of nitrous oxide and xenon, as well as the possible therapeutic implications in humans, are discussed. PMID- 14526228 TI - Transforming growth factor-beta 1-mediated neuroprotection against excitotoxic injury in vivo. AB - Ischemic preconditioning is a phenomenon that describes how a sublethal ischemic insult can induce tolerance to subsequent ischemia. This phenomenon has been observed after focal or global ischemia in different animal models. However, the hypothesis that bacterial infection might lead to neuronal tolerance to injury has not been investigated. To mimic cerebral bacterial infection, we injected bacterial lipopolysaccharide (LPS) in the right dorsal hippocampus, followed 24 hours later by an excitotoxic lesion using kainic acid in the mouse model. Quantification of lesion size after cresyl violet counterstaining revealed that LPS pretreatment afforded neuroprotection to CA3 neurons against KA challenge. To investigate the events underlying this protection, we studied the cytokine profile induced after LPS injection. Interleukin (IL)-1 beta and transforming growth factor beta 1 (TGF-beta 1) were the main cytokines expressed at 24 hours after LPS injection. Because IL-1 beta has been described as deleterious in acute injury, we decided to investigate the function of TGF-beta 1. An adenovirus expressing a constitutively active form of TGF-beta 1 was injected intracerebrally 1 week before the induction of excitotoxic lesion, and neuronal protection was observed. To confirm the neuroprotective role of TGF-beta 1, the TGF-beta 1 adenovirus was replaced by recombinant human TGF-beta 1 protein and total neuroprotection was observed. Furthermore, the antibody-mediated blocking of TGF-beta 1 action prevented the protective effect of pretreatment with LPS. We have demonstrated in vivo that the cerebral tolerance phenomenon induced by LPS pretreatment is mediated by TGF-beta 1 cytokine. PMID- 14526229 TI - Tacrolimus, a potential neuroprotective agent, ameliorates ischemic brain damage and neurologic deficits after focal cerebral ischemia in nonhuman primates. AB - Tacrolimus (FK506), an immunosuppressive drug, is known to have potent neuroprotective activity and attenuate cerebral infarction in experimental models of stroke. Here we assess the neuroprotective efficacy of tacrolimus in a nonhuman primate model of stroke, photochemically induced thrombotic occlusion of the middle cerebral artery (MCA) in cynomolgus monkeys. In the first experiment, tacrolimus (0.01, 0.032, or 0.1 mg/kg) was intravenously administered immediately after MCA occlusion, and neurologic deficits and cerebral infarction volumes were assessed 24 hours after the ischemic insult. Tacrolimus dose-dependently reduced neurologic deficits and infarction volume in the cerebral cortex, with statistically significant amelioration of neurologic deficits at 0.032 and 0.1 mg/kg and significant reduction of infarction at 0.1 mg/kg. In the second experiment, the long-term efficacy of tacrolimus on neurologic deficits and cerebral infarction was assessed. Vehicle-treated monkeys exhibited persistent and severe deficits in motor and sensory function for up to 28 days. A single intravenous bolus injection of tacrolimus (0.1 or 0.2 mg/kg) produced long lasting amelioration of neurologic deficits and significant reduction of infarction volume. In conclusion, we have provided compelling evidence that a single dose of tacrolimus not only reduces brain infarction but also ameliorates long-term neurologic deficits in a nonhuman primate model of stroke, strengthening the view that tacrolimus might be beneficial in treating stroke patients. PMID- 14526230 TI - DNA microarray analysis of hippocampal gene expression measured twelve hours after hypoxia-ischemia in the mouse. AB - Cell death from cerebral ischemia is a dynamic process. In the minutes to days after an ischemic insult, progressive changes in cellular morphology occur. Associated with these events is the regulation of competing programs of gene expression; some are protective against ischemic insult, and others contribute to delayed cell death. Many genes involved in these processes have been identified, but individually, these findings have provided only limited insight into the systems biology of cerebral ischemia. Attempts to characterize the coordinated expression of large numbers of genes in cerebral ischemia has only recently become possible. Today, DNA microarray technology provides a powerful tool for investigating parallel expression changes for thousands of genes at one time. In this study, adult mice were subjected to 30 minutes of hypoxia-ischemia (HI), and the hippocampus was examined 12 hours later for differential gene expression using a 15K high-density mouse EST array. The genomic response to HI is complex, affecting approximately 7% of the total number of ESTs examined. Assigning differentially expressed ESTs to molecular functional groups revealed that HI affects many pathways including the molecular chaperones, transcription factors, kinases, and calcium ion binding genes. A comprehensive list of regulated genes should prove valuable in advancing our understanding of the pathogenesis of cerebral ischemia. PMID- 14526231 TI - Differential effects of atrial natriuretic peptide on the brain water and sodium after experimental cortical contusion in the rat. AB - Atrial natriuretic peptide (ANP) plays an important role in the regulation of water and sodium in the body via cyclic GMP (cGMP) pathway. Although ANP has been shown to be protective in cerebral ischemia or intracerebral hemorrhage, its role in traumatic brain injury (TBI) has yet to be elucidated. We herein assessed ANP effects on brain water and sodium in TBI. Controlled cortical impact (3 mm depth, 6 m/sec) was used to induce an experimental cortical contusion in rats. Continuous administration of ANP 0.2 (n = 6) or 0.7 microg/kg/24 h (n = 6), cGMP analogue (8-Bromo-cGMP) 0.1 (n = 5) or 0.3 mg/kg/24 h (n = 5), or vehicle (n = 6) was begun 15 minutes after injury, using a mini-osmotic pump implanted into the peritoneal cavity. At 24 hours after injury, ANP significantly exacerbated brain edema in the injured hemisphere in a dose-dependent manner while it reduced brain sodium concentrations in both hemispheres. These ANP effects could be mimicked by a cGMP analogue. In the second series (n = 20), BBB integrity was assessed by evaluating the extravasation of Evans blue dye. ANP or cGMP analogue significantly worsened BBB disruption in the injured hemisphere at 24 hours after injury. These findings suggest that ANP administration exacerbates brain edema after the experimental cortical contusion in rats, possibly because of an increase in the BBB permeability via cGMP pathway, whereas it reduces brain sodium levels. PMID- 14526232 TI - Contributions of endothelial and neuronal nitric oxide synthases to cerebrovascular responses to hyperoxia. AB - Hyperoxia causes a transient decrease in CBF, followed by a later rise. The mediators of these effects are not known. We used mice lacking endothelial or neuronal nitric oxide synthase (NOS) isoforms (eNOS-/- and nNOS-/- mice) to study the roles of the NOS isoforms in mediating changes in cerebral vascular tone in response to hyperoxia. Resting regional cerebral blood flow (rCBF) did not differ between wild type (WT), eNOS-/- mice, and nNOS-/- mice. eNOS-/- mice showed decreased cerebrovascular reactivities to NG-nitro-L-arginine methyl ester (L NAME), PAPA NONOate, acetylcholine (Ach), and SOD1. In response to hyperbaric oxygen (HBO2) at 5 ATA, WT and nNOS-/- mice showed decreases in rCBF over 30 minutes, but eNOS-/- mice did not. After 60 minutes HBO2, rCBF increased more in WT mice than in eNOS-/- or nNOS-/- mice. Brain NO-metabolites (NOx) decreased in WT and eNOS-/- mice within 30 minutes of HBO2, but after 45 minutes, NOx rose above control levels, whereas they did not change in nNOS-/- mice. Brain 3NT increased during HBO2 in WT and eNOS-/- but did not change in nNOS-/- mice. These results suggest that modulation of eNOS-derived NO by HBO2 is responsible for the early vasoconstriction responses, whereas late HBO2-induced vasodilation depends upon both eNOS and nNOS. PMID- 14526233 TI - Cerebrovascular vasodilation to extraluminal acidosis occurs via combined activation of ATP-sensitive and Ca2+-activated potassium channels. AB - Albeit controversial, it has been suggested by several authors that nitric oxide (NO) serves as a permissive factor in the cerebral blood flow response to systemic hypercapnia. Potassium channels are important regulators of cerebrovascular tone and may be modulated by a basal perivascular NO level. To elucidate the functional targets of the proposed NO modulation during hypercapnia induced vasodilation, the authors performed experiments in isolated, cannulated, and pressurized rat middle cerebral arteries (MCA). Extracellular pH was reduced from 7.4 to 7.0 in the extraluminal bath to induce NO dependent vasodilation. Acidosis increased vessel diameter by 35 +/- 10%. In separate experiments, ATP sensitive potassium channels (KATP) were blocked by extraluminal application of glibenclamide (Glib), Ca2+-activated potassium channels (KCa) by tetraethylammonium (TEA), voltage-gated potassium channels (Kv) by 4 aminopyridine, and inward rectifier potassium channels (KIR) by BaCl2. Na+-K+-ATP ase was inhibited by ouabain. Application of TEA slightly constricted the arteries at pH 7.4 and slightly but significantly attenuated the vasodilation to acidosis. Inhibition of the other potassium channels or Na+-K+-ATP-ase had no effect. Combined blockade of KATP and KCa channels further reduced resting diameter, and abolished acidosis induced vasodilation. The authors conclude that mainly KCa channels are active under resting conditions. KATP and KCa channels are responsible for vasodilation to acidosis. Activity of one of these potassium channel families is sufficient for vasodilation to acidosis, and only combined inhibition completely abolishes vasodilation. During NO synthase inhibition, dilation to the KATP channel opener pinacidil or the KCa channel opener NS1619 was attenuated or abolished, respectively. The authors suggest that a basal perivascular NO level is necessary for physiologic KATP and KCa channel function in rat MCA. Future studies have to elucidate whether this NO dependent effect on KATP and KCa channel function is a principle mechanism of NO induced modulation of cerebrovascular reactivity and whether the variability of findings in the literature concerning a modulatory role of NO can be explained by different levels of vascular NO/cGMP concentrations within the cerebrovascular tree. PMID- 14526234 TI - Energy dysfunction as a predictor of outcome after moderate or severe head injury: indices of oxygen, glucose, and lactate metabolism. AB - The purpose of this study was to determine if the relationship between abnormalities in glucose, lactate, and oxygen metabolism were predictive of neurologic outcome after moderate or severe head injury, relative to other known prognostic factors. Serial assessments of the cerebral metabolic rates for glucose, lactate, and oxygen were performed using a modified Kety-Schmidt method. In total, 31 normal control subjects were studied once, and 49 TBI patients (mean age 36+/-16 years, median GCS 7) were studied five times median per patient from postinjury days 0 to 9. Univariate and multivariate analyses were performed. Univariate analysis showed that the 6-month postinjury Glasgow Outcome Scale (GOS) was most strongly associated with the mean cerebral metabolic rate of oxygen (CMRO2) (P = 0.0001), mean arterial lactate level (P = 0.0001), mean arterial glucose (P = 0.0008), mean cerebral blood flow (CBF), (P = 0.002), postresuscitation GCS (P = 0.003), and pupillary status (P = 0.004). Brain lactate uptake was observed in 44% of all metabolic studies, and 76% of patients had at least one episode of brain lactate uptake. By dichotomized GOS, patients achieving a favorable outcome (GOS 4-5) were distinguished from those with an unfavorable outcome (GOS1-3) by having a higher CMRO2 (P = 0.003), a higher rate of abnormal brain lactate uptake relative to arterial lactate levels (P = 0.04), and lesser degrees of blood-brain barrier damage based on CT findings (P = 0.03). CONCLUSIONS: During the first 6 days after moderate or severe TBI, CMRO2 and arterial lactate levels are the strongest predictors of neurologic outcome. However, the frequent occurrence of abnormal brain lactate uptake despite only moderate elevations in arterial lactate levels in the favorable outcome patients suggests the brain's ability to use lactate as a fuel may be another key outcome predictor. Future studies are needed to determine to what degree nonglycolytic energy production from alternative fuels such as lactate occurs after TBI and whether alternative fuel administration is a viable therapy for TBI patients. PMID- 14526253 TI - [Neurosurgical embryology. Part 1: Cell differentiation]. AB - In pluricellular organisms, cell differentiation helps to decrease the total amount of energy needed for life. These differentiations can be evidenced at the tissular, the cellular or the molecular levels. Cell differentiation is a progressive process achieved during embryogenesis; different steps in the program can be described. One of the explanations to account for cell differentiation is the specific expression of proteins, called transcription factors, that can control the expression of selected genes. These factors are classified according to their biochemical pattern allowing description of several families of transcription factors. One of the salient questions during embryogenesis is to understand the mechanisms involved in cell differentiation. The first event is due to asymmetry of mitosis leading to the generation of two cell lineages. This is favored by the initial ovocyte polarization. The second event is due to cell interactions (namely inductions). These inductions may be explained either by cell-cell contact (favored by cell adhesion molecules) or by secreting factors that can be either hydrophilic or lipophilic. PMID- 14526254 TI - [Neurosurgical Embryology. Part 2: Recent data on normal and pathological development of the cortex]. AB - Understanding how the cortex develops has gained very important new data thanks to both experimental and clinical studies. Experimental studies have shown that: -neurons are generated in the ventricular zone by asymmetric mitoses; --the first cortical region to differentiate is the so-called pre-plate that plays a major role in the control of neuroblasts migration; --neuroblasts arise according to an inside-outside gradient; they migrate either along the processes of radial cells or according to a new type of non radial migration; --all the cortical neurons are not generated by the dorsal telencephalon; some of these neurons derive from the basal telencephalon; --neuroblasts acquire their specificity during their way to the cortical plate. There are several genetic syndromes leading to a malformation of the cortex. Classic lissencephaly is essentially due to mutations of the LIS1 or the DCX genes. These genes code for proteins that are involved in cytoskeleton functions. Reelin is responsible for a human syndrome associating pachygyria and cerebellar hypotrophy. Subventricular heterotopia can be X- inherited and are due to a mutation of the gene coding for filamin 1. PMID- 14526255 TI - [Neurosurgical Embryology. Part 3: Molecular control of corpus callosum development]. AB - The corpus callosum is the most important cerebral commissure allowing axonal fibres to cross the midline. Corpus callosum agenesis is an important condition in man that can reveal numerous genetic syndromes. The corpus callosum develops from the commissural plate, a dorsal region of the telencephalon. Then, axons growing from pyramidal neurons of cortical layer III extend and cross the midline. In experimental models, it is possible to decipher two conditions in which the development of the corpus callosum is impaired. The first condition is characterized by an impairment of the formation of the roof of the telencephalon (the primordium of the commissural plate). This condition can be explained by an abortive induction of this region by an impairment of BMP signaling. This can generate all the forms of holoprosencephaly. Other forms are due to a defective gene coding Hesx1, a transcription factor involved in the control of telencephalic morphogenesis. Such a genetic defect can be observed in human dominant forms of septo-optic dysplasia. The second condition is explained by an impairment of the molecular control of axon growth: such is the case for the couple netrin 1 and DCC or for the adhesion molecule L1. PMID- 14526257 TI - [Endonasal endoscopic access to the base of the skull]. PMID- 14526256 TI - [Neurosurgical embryology. Part 4: What are stem-cells?]. AB - Stem-cells have been identified in the adult human brain in two zones which are the subventricular zone and the gyrus dentatus of the hippocampus. Improvement of techniques aimed to identify, to localize and to follow the lineage of these cells have been crucial to the understanding of the following processes: a) identification of cellular proliferation, b) specific immunostaining of differentiated glial and neuronal cells, c) transplantations to decipher between intrinsic stem-cell properties and influence of the environment on the fate of the cell. Furthermore, it seems that stem-cells from other sources than the brain can differentiate into neurons both in vitro and in vivo. The aim of this review is to sum up what is known about cerebral stem-cells and the challenging tools they mean for the future. PMID- 14526259 TI - [Inter-regional visioconference teaching model]. PMID- 14526258 TI - [Anti-glioma vaccination]. PMID- 14526261 TI - Cardiovascular risk in type 2 diabetics and pharmacological regulation of mealtime glucose excursions. AB - In type 2 diabetic patients mealtime glucose fluctuations are important determinants of overall glucose control and overall risk of diabetes cardiovascular complications. In fact, acute elevation of plasma glucose concentrations trigger an array of tissue response that may contribute to development of such vascular complications since it may result in a thrombophilic condition, causes endothelial dysfunction (possibly through a reduction of nitric oxide availability) and is responsible for non-enzymatic glycation and production of free- radicals with ensuing oxidative stress. To keep post-prandial glucose with narrow range, metiglinide analogues drugs have been developed. In particular, repaglinide and nateglinide seem the most useful ones. In fact, both drugs improve 1(st) phase insulin release but they do not affect the total daily amount of insulin released by the pancreas. Due to the mechanism of action and to pharmacokinetic properties, repaglinide and nateglinide allow diabetic patients to get a more tight metabolic glucose control with a contemporary reduction in the cases of severe hypoglycaemia. In conclusions, repaglinide and nateglinide are new and powerful pharmacological tools not only for achieving a better metabolic glucose control but also for preventing the development of diabetes related cardiovascular complications. PMID- 14526262 TI - Reproducibility of plasma insulin kinetics during intraperitoneal insulin treatment by programmable pumps. AB - OBJECTIVE: To study the reproducibility of plasma insulin kinetics during intraperitoneal (IP) insulin therapy using an implanted programmable pump in patients with type 1, insulin dependent, diabetes mellitus (IDDM). RESEARCH DESIGN AND METHODS: In a group of ten type 1 IDDM patients beginning chronic IP insulin treatment with an implanted pump, plasma free insulin profiles were determined from 12: 00 am to 12: 30 pm on two separate test days, one month apart. Anti-insulin antibody (AIA) levels were measured on each test day. RESULTS: From test day 1 to test day 2, no difference was observed in morning fasting free insulin levels (m +/- SD): 9.7 +/- 5.4 mU/L versus 9.8 +/- 5.3 mU/L, insulin peak values: 19.1 +/- 17 mU/L versus 20.8 +/- 9.9 mU/L, time to peak: 40 +/- 15 versus 42.8 +/- 16 minutes or post-bolus area under the plasma free insulin curve (AUC): 40.7 +/- 29 mU/L.h versus 45.5 +/- 29 mU/L.h. The intrapatient coefficient of variation was 14.4 +/- 13% for insulin peaks and 16.9 +/- 9.2% for post-bolus AUC. A significant increase in AIA levels (m +/- SD) was observed from 16.5 +/- 18% on test day 1 to 28.1 +/- 28% on test day 2. CONCLUSIONS: The reproducibility of plasma free insulin profiles is highly satisfactory during insulin delivery by the IP route using an implanted device. PMID- 14526263 TI - Influence of plasma amino acid level on vasopressin secretion. AB - OBJECTIVES: Vasopressin (VP) is known to be elevated in patients with diabetes mellitus (DM). While the influence of acute hyperglycemia has been ruled out, the mechanism or the osmotically active compound responsible for the increase in VP secretion is still not elucidated. Because the plasma level of several amino acids (AAs) is increased in DM, we evaluated whether AAs could represent an effective osmotic stimulus for VP secretion. RESEARCH DESIGN AND METHODS: In a cross-over study, eight healthy volunteers randomly received an infusion of isotonic saline (control) or mixed AA solution, i.v., at a low or a high rate (2 or 4.5 mg/min/kg BW, respectively). Plasma VP (P(VP)) was measured for two hours before and three hours during AA or control infusion. RESULTS: AA infusion induced a dose-dependent elevation in plasma AA concentration but did not alter P(VP). However, effective plasma osmolality (P(osm)) (osmolality minus urea concentration) remained unchanged because a concommittant fall in plasma sodium concentration (P(Na)), likely due to sodium-linked uptake of AA in peripheral cells, compensated for the rise in plasma AA. CONCLUSION: The stability of effective P(osm) may explain the lack of change observed in P(VP). Because sodium is a very efficient stimulus for VP secretion, it may be assumed that the fall in P(Na) occurring during AA infusion should have reduced VP secretion and thus P(VP). In this setting, the stability of P(VP) suggests that AAs induced an increase in VP secretion which counterbalanced the fall attributable to the decrease in P(Na). In conclusion, in acute experiments, AAs seem to represent an effective stimulus for VP secretion, almost equally potent as sodium. Further studies are needed to evaluate their contribution to the high P(VP) seen in the chronic setting of DM. PMID- 14526264 TI - Low rate of glucose 6-phosphate hydrolysis in liver cells is a physiological feature of non-diabetic wild sand rats (Psammomys obesus). AB - OBJECTIVE: In this study we have compared glucose metabolism and liver gluconeogenesis in wild adult desert gerbil Psammomys obesus fed with their natural halophilic plants and Wistar rats fed on a laboratory chow. Psammomys obesus is a natural model of insulin resistance when fed a rodent laboratory chow. METHODS: Basal glucose and insulin were determined in plasma of fasting animals. Hepatocyte gluconeogenesis from lactate-plus-pyruvate was investigated in perifused hepatocytes by assessing simultaneously glucose synthesis rate and intracellular oxaloacetate, phosphoenolpyruvate, 3-phosphoglycerate, fructose 6 phosphate and glucose 6-phosphate (G6P) under true steady state conditions. RESULTS: Fasting blood glucose (2.8 +/- 0.1 vs 4.8 +/- 0.4 mmol.L(- 1)) and plasma insulin concentration (129 +/- 14 vs 150 +/- 21 pmol.L(-1)) were significantly lower in Psammomys as compared to albino rats. Maximal gluconeogenic rate was also lower in Psammomys (2.3 +/- 0.3 vs 5.1 +/- 0.3 micromol x min(-1) x g dry cells(-1)). This effect was related to a slower hydrolysis of G6P. CONCLUSION: A lower G6P hydrolysis in Psammomys as compared to wistar was the main difference between the two groups of liver cells. Such feature may represent the major metabolic adaptation permitting Psammomys to survive despite its severe restrictive natural conditions. Indeed, a low G6P hydrolysis allows an insulin resistance state, with a high lipogenic activity, but associated with low blood glucose. The rise in blood glucose occurring when Psammomys are fed with exogenous carbohydrates perturbs such delicate metabolic equilibrium, resulting thus in a diabetic state because of the deleterious effect of hyperglycemia. PMID- 14526265 TI - Comparative performance of serum cystatin-c versus serum creatinine in diabetic subjects. AB - OBJECTIVE: Serum cystatin-C, a protein with constant production rate, undergoes glomerular filtration. Cystatin-C is a candidate surrogate marker, allegedly superior to serum creatinine, for estimating glomerular filtration rate, due to its high correlation with absolute measurement of the latter. The aim of this study was to assess from intra- and inter-subject variability the performance of cystatin-C in a cohort of diabetic patients spanning a wide range of kidney function and to compare it to that of serum creatinine. RESEARCH DESIGN AND METHODS: 98 consecutive diabetic in-patients (45 type 1 and 53 type 2) were included. Mean age was 53 +/- 15 years (1SD). Creatinine clearance was 98 ml/min (median; range: 16-244). We used the discriminant ratio (DR) methodology to compare the performance of serum cystatin-C vs. that of creatinine to segregate subjects according to their glomerular filtration rate. RESULTS: Serum creatinine values on day 1 and 2 were 1.10 +/- 0.76 and 1.07 +/- 0.89 mg/dl and concentrations of cystatin-C were 1.10 +/- 0.60 and 1.06 +/- 0.63 mg/L. A close linear relationship was observed between means of duplicates for creatinine and cystatin-C (Pearson product-moment correlation 0.92). DR was obtained from the ratio of the underlying between-subject to the within-subject standard deviations. DR values were 5.23 for creatinine and 8.82 for cystatin-C (P<0.0001), implying superior discriminating ability for cystatin-C. Once adjusted for attenuation, measured Pearson product-moment correlation rose from 0.92 to 0.97. The DR methodology allowed for deriving an unbiased linear regression equation between methods, with slope and intercept at 0.79 and 0.23, respectively. CONCLUSIONS: Serum cystatin-C better discriminates among a population of type 1 and 2 diabetic patients with regard to their estimated glomerular filtration rate when compared with conventional serum creatinine measurement. PMID- 14526266 TI - Tranilast attenuates vascular hypertrophy, matrix accumulation and growth factor overexpression in experimental diabetes. AB - OBJECTIVES: The growth factors transforming growth factor-B (TGF-B) and epidermal growth factor (EGF) have both been implicated in the hypertrophic structural changes in the vasculature that are characteristic features of both human and experimental diabetes. Recently, tranilast (N(3,4-dimethoxycinnamoyl)anthranilic acid), a drug used in the treatment of allergic and dermatological diseases, has also been reported to inhibit transforming growth factor-B (TGF-B)-mediated collagen formation. However, its effects on vascular hypertrophy in diabetes are unknown. The present study thus sought to determine the effects of tranilast on both TGF-B and EGF expression and mast cells in mediating the trophic vascular changes in experimental diabetes. METHODS: Vessel morphology, growth factors and collagen gene expression and matrix deposition were examined in the mesenteric arteries of control rats treated with or without tranilast, and streptozotocin induced diabetic Sprague-Dawley rats treated with or without tranilast (200 mg/kg/day) during a 3-week period. RESULTS: Compared with control animals, diabetic rats had significantly increased vessel weight, wall: lumen ratio, ECM accumulation, gene expression of TGF-B1, EGF, and both alpha1 (I) and alpha1 (IV) collagen. Tranilast treatment did not influence plasma glucose or systemic blood pressure. However, tranilast significantly reduced mesenteric weight, wall: lumen ratio and matrix deposition and also attenuated the overexpression of TGF-B1, EGF, and both alpha1 (I) and alpha1 (IV) collagen mRNA in diabetic rats. CONCLUSION: These findings indicate that tranilast ameliorates pathological vascular changes observed in experimental diabetes in association with reduced growth factor expression independent of blood glucose or systemic blood pressure. PMID- 14526267 TI - Description of microangiopathy in children with type 1 diabetes mellitus: a 19 year retrospective study. AB - BACKGROUND: The aim of the study was to estimate the outcome of patients with type 1 diabetes followed in a university hospital in the paediatric department and then in the adult diabetic department for at least 10 years. METHODS: We made a retrospective analysis of 50 patients (28 women and 22 men) with type 1 diabetes with disease duration of 19 +/- 6 years and analysed whether retinopathy and nephropathy had progressed, had remained unchanged or had improved or normalised. RESULTS: The mean age of diabetes onset was 8 +/- 4 years (1-16). Ketoacidosis revealed diabetes in 36% of the children. Mean HbA(1c) was 8.6 +/- 1.8%, and was over 8.5% in 34% of the patients. The mean age at onset of puberty (Tanner stage II) was 12 +/- 1 years in girls and 13 +/- 1 years in boys. Mean HbA(1c) was 7.9 +/- 1.2% during the year before onset of puberty and 8.7 +/- 1.1% in the following 3 years, corresponding to a 10% pubertal increase in HbA(1c). Retinopathy was seen in 50% of the patients at a mean age of 22 +/- 5 years, 15 +/- 6 years after onset of diabetes. Mean HbA(1c) was 9.7 +/- 1.6% in patients with proliferative retinopathy, 9.0 +/- 1.5% in patients with non proliferative retinopathy, and 8.1 +/- 1.3% in those without (p=0.02, proliferative versus no retinopathy, p > 0.05 non proliferative versus no retinopathy). Microalbuminuria was diagnosed in 26% of the patients. Mean HbA(1c) was 9.3 +/- 2.1% in patients with microalbuminuria versus 8.1 +/- 1.3% in those with normoalbuminuria (p=0.02). CONCLUSIONS: Glycemic control was similar in patients with non proliferative retinopathy and those without. Proliferative retinopathy and nephropathy were both related to the level of glycemic control. PMID- 14526268 TI - Acute-phase proteins among patients with type 1 diabetes. AB - OBJECTIVE: To determine whether young type 1 diabetic patients without clinical microvascular or macrovascular complications have altered levels of acute-phase proteins (AFP), alpha(1)-acid glycoprotein (AGP), C-reactive protein (CRP) and fibrinogen and whether their AFP levels are related to glycemic control. RESEARCH DESIGN AND METHODS: We studied cross-sectionally 48 type 1 diabetic outpatients (25 males) aged 19.9 +/- 9.8 years with a duration of diabetes of 5 (1-21) years, without clinical chronic complications and 66 non-diabetic subjects (26 males) aged 23.1 +/- 10.9 years. Inclusion criteria were normoalbuminuria, normal eye fundoscopy, and no evidence of cardiovascular disease or neuropathy. RESULTS: High CRP [0.23 (0.01-2.90 l) vs (0.14 (0.01-2.41l) mg/dl p=0.01] and AGP [53.5(40 78) vs 40.0(40-115) mg/dl p=0.0001] levels were found in patients with type 1 diabetes compared to nondiabetic subjects. In the pooled group studied, AGP was correlated with CRP, HbA(1c), fasting plasma glucose (FBG) and AER and CRP was correlated with HbA(1c) and AER. The correlation of AGP and CRP with AER persisted after controlling for HbA(1c) and FBG. Stepwise multiple regression with AGP as the dependent variable showed that FBG and HbA(1c) were the significant independent variables. No correlation between AFP and HBA(1c) and FBG was observed in the diabetic group. CONCLUSIONS: According to our results, AFP, a known marker of low-grade chronic inflammation, are increased in patients with type 1 diabetes probably independently of glycemic control and the presence of clinical microvascular or macrovascular disease. The influence of AFP on the development of chronic complications in patients with type 1 diabetes must be addressed in prospective studies. PMID- 14526269 TI - Effects of dipeptide administration on hypoglycaemic counterregulation in type 1 diabetes. AB - OBJECTIVES: To investigate if a dipeptide made of glutamine and alanine is able to contribute to the recovery from insulin-induced hypoglycaemia in type 1 diabetes. RESEARCH DESIGN AND METHODS: Fifteen adult type 1 patients were randomly assigned to study group (n=7): intravenous infusion of 20 g Dipeptiven in normal saline (i.e., 8 g alanine and 13 g glutamine), or control group (n=8): same infusion, normal saline only. A 150 min gradual hypoglycaemic hyperinsulinemic clamp was administered after 2 h of infusion. Counterregularory hormones, symptoms, and cognitive function (4 choice reaction test) were regularly measured during the study. RESULTS: Blood glucose and glucose infusion rates were similar in the 2 groups. Circulating levels of alanine and glutamine peaked at 90 min and remained elevated throughout the test. This was associated with significant differences in: glucagonemia 107 +/- 20 vs 58 +/- 8 pg/ml, and neuroglycopenic symptoms scores: 7 +/- 3 vs 18 +/- 13, at t 150 min, in study and control group, p<0.05. Dysautonomic symptoms, cognitive tests as well as epinephrine, norepinephrine, cortisol and growth hormone were similar between groups. CONCLUSION: Intravenous infusion of a dipeptide made of alanine and glutamine is capable to reactivate glucagon secretion during insulin-induced hypoglycaemia and to reduce hypoglycaemic symptoms. PMID- 14526270 TI - Interest of pronostic score for optimal clinical management of obese patients. AB - OBJECTIVE: The aim of this retrospective study was to identify key factors favoring weight regain. METHODS: Therefore, prognostic factors for weight loss and regain have been retrieved and identified among 186 obese patients (40 +/- 2 y and 34 +/- 0.3 kg/m(2)) through the use of a simple questionnaire which considered psychological issues such as depression, anxiety and the patient's individual capacity to follow a diet. All patients have been prescribed a low calorie diet (1200 kcal/day) being either balanced, food-combining or low-CHO. Follow-up averaged 6.4 +/- 0.3 months of which weight loss accounted for 4.4 +/- 0.1 months. RESULTS: Weight loss was similar in all 3 groups and averaged 6.0 +/- 0.2 kg. The final weight loss and weight regain results have been identical for the 3 groups. A high/bad prognostic score rating is associated with weight regain (P<0.02). Weight regain is significantly influenced by the initial weight loss speed (P<0.0001). CONCLUSIONS: The proposed prognostic score is likely to be helpful for clinicians better to profile the therapeutic approach to individualized obesity management and follow-up. Diet composition does not influence either weight loss or weight regain. PMID- 14526271 TI - Hepatic expression of the human insulin gene reduces glucose levels in vivo in diabetic mice model. AB - OBJECTIVES: The objective of these studies was to evaluate human insulin gene expression following intraliver plasmid injection in diabetic mice as a potential approach to gene therapy for insulin-dependent diabetes mellitus. METHODS: The fragment containing human proinsulin gene lacking its own promoter, was cloned into plasmids containing promoter and enhancer of cytomegalovirus or human hepatitis B virus. The resulting gene constructs were first tested in vitro using 3T3 fibroblast cell line and subsequently in vivo applying streptozotocin-induced diabetic mice. RESULTS: We found significant reduction in glucose levels in both experimental systems, giving evidence that prolonged constitutive systemic secretion of bioactive human (pro)insulin has been attained in non-neuroendocrine cell line in vitro and in mice following intra-liver plasmid injection. CONCLUSION: Our data demonstrate the reduction of glucose levels in vitro in 3T3 fibroblast cells and in vivo in diabetic mice after treatment with plasmids expressing proinsulin, giving evidence that those constructs may have certain usage also in human gene therapy of diabetes mellitus type 1. PMID- 14526272 TI - Time-action profile of the long-acting insulin analogue insulin glargine in comparison to NPH insulin in Japanese volunteers. PMID- 14526273 TI - Allergy to insulin in a woman with gestational diabetes mellitus: transient efficiency of continuous subcutaneous insulin lispro infusion. AB - We report the case of a 31-year-old pregnant woman. She required insulin for the treatment of gestational diabetes from 27 weeks of amenorrhoea to delivery. An allergy to insulin was suspected because she presented with local symptoms at insulin injection sites and a decrease in efficiency of insulin. This diagnostic was confirmed by skin-prick tests. A treatment with subcutaneous continuous lispro insulin analogue infusion was initiated with an oral antihistaminic drug without local reaction. Seven weeks after the initiation of insulin pump, local reactions reappeared. The insulin analogue lispro is not always an alternative in insulin allergy. However, in the case we report, the lack of allergy during a few weeks allowed the birth of a normal infant. PMID- 14526274 TI - Diabetes mellitus in Turkish immigrants in Sweden. AB - This study analyses the risk of self-reported diabetes mellitus among Turkish born immigrants in Sweden. Two simple random samples were used: The Swedish National Board of Health and Welfare Immigrant Survey, and the Swedish Survey of Living Conditions, both from 1996. Totally 526 Turkish immigrants, 285 men and 241 women, were compared with 2854 Swedish controls, 1425 men and 1429 women, all in ages 27-60 years. Data were analysed by sex in an age-adjusted model, and a full model also included education, employment status, BMI and country of birth (logistic regression). Among Turkish men, age-adjusted diabetes prevalence was not higher than among Swedish men, odds ratio (OR) 1.04 (95% confidence interval (CI) 0.35-3.11). Among Turkish women, age-adjusted diabetes prevalence was higher than among Swedish women, OR 3.22 (95% CI 1.36-7.64), but when also adjusting for educational level, employment status and BMI, OR was 1.22 (95% CI 0.41-3.66). We conclude, that age-adjusted presence of known diabetes was higher among Turkish born women than among Swedish women, but was explained by lower employment rate, lower educational status and a higher level of overweight and obesity. PMID- 14526277 TI - WHI trial: it's time to be critical! PMID- 14526275 TI - Home blood pressure monitoring in diabetic population. AB - Normalization of blood pressure is today as necessary as an optimal blood glucose control. These targets aim at reducing the negative consequences of hypertension which is particularly frequent and of poor prognosis for these patients. Nevertheless, the definition of the real blood pressure of these patients is uneasy because of the lack of precision of clinical blood pressure measurement. New methods have been developed to address this insufficiency. Home blood pressure monitoring is characterised by good compliance and accuracy, but patient education needs a special training and validated automated devices. This method has to be proposed to the diabetic population as soon as hypertension may be a problem. These patients will contribute to monitor their own blood pressure as they do it with blood glucose control. PMID- 14526278 TI - WHI: Now that the dust has settled: a commentary. PMID- 14526280 TI - Work hours for practicing obstetrician-gynecologists: the reality of life after residency. AB - OBJECTIVE: Recently the Accreditation Council for Graduate Medical Education placed restrictions on all residency programs that limited work hours to 80 hours per week. The objective of this study was to determine the work hours for practicing obstetrician-gynecologists in an urban center. STUDY DESIGN: A questionnaire about physician demographics and work hours was mailed to all obstetrician-gynecologists in Houston in June 2002. Chi(2) testing was used for statistical analysis. RESULTS: One hundred eighty-nine surveys were mailed. One hundred surveys (56%) were returned complete and analyzed. Sixty-two physicians reported working >80 hours per week. Physicians were more likely to work >80 hours per week if they were men or if they were >50 years old. Marital status and having children living at home did not affect work hours. CONCLUSION: Most obstetrician-gynecologists in Houston work longer hours than the number of hours that is imposed by the Accreditation Council for Graduate Medical Education resident work hour guidelines. PMID- 14526279 TI - Job satisfaction among program directors in obstetrics and gynecology: a national portrait. AB - OBJECTIVES: The purpose of this study was to determine job satisfaction among program directors in obstetrics and gynecology with the use of a validated tool and to identify specific sources of dissatisfaction that might lead to job change. STUDY DESIGN: The program director satisfaction and a global job satisfaction survey were sent to all program directors in the United States. Motivators for seeking a job change were assessed. The chi(2) test, Kruskal Wallis test, correlation analysis, and multiple linear regression were used. RESULTS: Seventy percent of 254 surveys were completed. Global job satisfaction (minimum, 4; maximum, 16) was 11.9+/-2.9; mean program director satisfaction score was 135+/-25.8 (minimum, 54; maximum, 200). Job satisfaction was highest in chairs, full professors, those whose age was >50 years, and those with >5 years of experience (P=.02) and in facets that were related to work with residents, colleagues, and patients. Dissatisfaction was highest with regard to salary, promotion opportunities, and resources. Forty-six percent of those who responded were considering a job change in 3 years; the most common reason for a job change that was cited was administrative hassles. CONCLUSION: Although job satisfaction is high among program directors, administrative hassles may lead to high rate of rapid turnover. PMID- 14526281 TI - Obtaining patient permission for student participation in obstetric-gynecologic outpatient visits: a randomized controlled trial. AB - OBJECTIVES: Our purpose was to compare a scripted verbal query with a detailed written permission slip in obtaining patient satisfaction and permission for student involvement in outpatient obstetrics-gynecologic visits. STUDY DESIGN: A prospective, randomized, controlled study was performed using a questionnaire to compare current practice to the study groups. The chi(2) test was used to calculate P values; P<.05 was considered significant. RESULTS: Patient demographics and satisfaction were similar among the three groups: 86% of controls and 79% of study groups agreed to student participation (P=.056). All preferred having the nurse ask permission (86% vs 86%) versus the physician (34% vs 25%) or the student (6% vs 3%). Permission was independent of student gender, visit purpose, or previous exposure to students. CONCLUSION: Patients want a nonphysician to ask permission for student participation independent of method of request, visit purpose, student gender, or previous experience with students. Physician or student requests for consent may unduly influence participation. PMID- 14526282 TI - What influences medical students to pursue careers in obstetrics and gynecology? AB - OBJECTIVES: The purpose of this study was to identify factors that influence medical students to choose of obstetrics/gynecology as a career specialty. STUDY DESIGN: A Web-based survey of medical students was conducted that included demographics, desire to enter the obstetrics and gynecology specialty, factors that influence selection of the obstetrics and gynecology specialty, perceptions about the obstetrics and gynecology specialty, and incidents of encouragement and discouragement toward entering the obstetrics and gynecology specialty. Analysis included parametric and nonparametric testing. RESULTS: Fifty-one percent of the students (n=205) returned the survey: 131 women (64%) and 72 men (35%); 2 respondents did not specify gender. Various factors influenced a medical student to pursue a career in obstetrics and gynecology, which revealed some specific areas of significant difference between genders, particularly in relation to lifestyle issues and practice trends. CONCLUSIONS: This study amplifies the importance of understanding the role of student gender when exploring the obstetrics and gynecology specialty as a career choice. Appreciating factors that influence decisions to enter the obstetrics and gynecology specialty provides opportunities to impact the experiences and decisions of students toward considering the obstetrics and gynecology specialty as a career choice. PMID- 14526283 TI - Enhancing resident training in abortion and contraception through hospital community partnership. AB - OBJECTIVES: The study was undertaken to assess the efficacy of a hospital community rotation in abortion and contraception. STUDY DESIGN: The program was developed and implemented by the Planned Parenthood League of Massachusetts (PPLM) and the Department of Obstetrics and Gynecology at Baystate Medical Center in November 2000. Abortion and contraceptive training is integrated into postgraduate year 3 and includes didactic presentations and 10 mandatory clinical sessions in first trimester abortion care. RESULTS: Five residents complete the program annually. Before rotation, residents report their competence in first trimester abortion procedure as 2 on a scale of 1 to 5 (1=not at all comfortable, 5=very comfortable). After rotation, the reported competency rating is 4.5 on the same scale. Residents, trainers, and staff report high satisfaction with the program. CONCLUSION: A collaborative hospital-community rotation in abortion and contraception increases residents' clinical competence in these areas and is highly acceptable to residents, faculty, and staff. PMID- 14526284 TI - A comprehensive pelvic dissection course improves obstetrics and gynecology resident proficiency in surgical anatomy. AB - OBJECTIVE: This study was undertaken to evaluate the impact of a pelvic dissection course on resident proficiency in surgical anatomy. STUDY DESIGN: Over a 1-year period, residents attended a course consisting of pretesting and posttesting, lectures, and pelvic dissection. Tests results were analyzed using paired Student t test, analysis of variance, and Kruskal-Wallis statistics. RESULTS: Of 42 residents, 24 completed all testing (study cohort). On written and practical examinations, resident scores improved a median of 42% and 29% (both P<.0001). Postgraduate year (PGY) 2 demonstrated the greatest improvement on the practical and PGY-3s demonstrated the greatest improvement on the written. Baseline written and practical results discriminated PGY level (construct validity): PGY-2=PGY-3or=20% difference in birth weight, relative to the larger twin. Fisher's exact, Mann Whitney U, and Spearman rho tests were used for statistical analysis. RESULTS: Of 130 twin pregnancies, 16 pregnancies (12.3%) had discordant birth weight. Crown-rump length disparity was correlated positively with birth weight discordance (r=0.31; P<.001). Of pregnancies with a discrepancy of 3 days discrepancy (P=.004), which resulted in a likelihood ratio of 5.9 for having discordant birth weight. CONCLUSION: Twins who are ultimately discordant at birth may exhibit differences in growth as early as 11 to 14 weeks of gestation. PMID- 14526299 TI - A randomized comparison of two regimens of misoprostol for second-trimester pregnancy termination. AB - OBJECTIVE: The purpose of this study was to compare the efficacy and side effects of two different misoprostol regimens for second-trimester pregnancy termination. STUDY DESIGN: We performed a randomized clinical trial in patients who were at 14 to 23 weeks of gestation and who were admitted for medical termination of pregnancy. All patients received 800 microg of vaginal misoprostol and were assigned randomly to 400 microg of oral misoprostol or 400 microg of vaginal misoprostol every 8 hours. Efficacy and side effects were compared. The mean induction time of the study group was compared with that of an historic control group that had received 400 microg vaginally every 12 hours. RESULTS: Forty-three women were assigned randomly, 22 women to vaginal misoprostol and 21 women to oral misoprostol. Induction time and hospital stay were slightly shorter for the oral group; however, the differences were not significant. Side effects were similar for both groups. CONCLUSION: After an initial 800 microg dose of vaginal misoprostol, a regimen of 400 microg of oral misoprostol every 8 hours is as effective as the same dose of vaginal misoprostol with no additional side effects, which provides a convenient alternative for midtrimester pregnancy termination. PMID- 14526300 TI - Vaginal birth after cesarean section: trial of labor or repeat cesarean section? A decision analysis. AB - OBJECTIVE: The risk of perinatal death associated with labor after previous cesarean section appears higher than with a repeated cesarean section. On the other hand, repeated cesarean sections are associated with increased maternal morbidity and mortality from placental pathologic conditions (previa or accreta) on subsequent pregnancies. The study was undertaken to analyze the decision for a trial of labor or a repeated cesarean section, after a prior cesarean section, with varying desire for an additional pregnancy. STUDY DESIGN: A model was formulated using a decision tree, based on the reported risks of the two approaches. Sensitivity analysis was performed over a variety of probabilities (eg, chance of uterine rupture or neonatal death, chance of rescue cesarean section, desire for an additional pregnancy) and utilities (eg, use of hysterectomy or neonatal death). RESULTS: The model favors a trial of labor if it has a chance of success of 50% or above and if the wish for additional pregnancies after a cesarean section is estimated at near 10% to 20% or above because the delayed risks from a repeated cesarean section are greater than its immediate benefit. The model was robust over a wide range of assumptions. CONCLUSION: An optimal decision for a trial of labor or a repeated cesarean section is substantially determined by the wish for future pregnancies. The default option of a repeated cesarean section is not directly applicable in populations in which family planning often extends over two children. PMID- 14526301 TI - Oral misoprostol for premature rupture of membranes at term. AB - OBJECTIVE: The study was undertaken to compare the efficacy, safety, and maternal satisfaction of oral misoprostol and intravenous oxytocin for labor induction in women with premature rupture of membranes at term. STUDY DESIGN: One hundred five women were stratified by parity and randomly assigned to oral misoprostol 75 microg every 4 hours as needed to establish labor or to intravenous oxytocin. RESULTS: The induction to vaginal delivery time with oral misoprostol was 737 (+/ 426) minutes compared with 573 (+/-318) minutes with oxytocin (P=.04). The incidence of hyperstimulation was lower in the misoprostol group (6.0% vs 27.1%, P=.005). Women were more likely to be very satisfied with their care in the misoprostol group (86.0% vs 63.4%, P=.02). CONCLUSION: In women at term with premature rupture of membranes, oral misoprostol resulted in a longer induction to vaginal delivery interval but increased maternal satisfaction and less hyperstimulation compared with intravenous oxytocin. Further research is needed to assess uncommon neonatal and maternal outcomes. PMID- 14526302 TI - Intrapartum risk factors for permanent brachial plexus injury. AB - OBJECTIVE: The purpose of this study was to compare maternal, neonatal, and second stage of labor characteristics in shoulder dystocia deliveries that result in permanent brachial plexus injury with shoulder dystocia deliveries that result in no injury. STUDY DESIGN: Our cases were culled from a database of deliveries that resulted in permanent brachial plexus injuries and matched to control cases that were taken from a database of consecutive shoulder dystocia deliveries from one hospital. Deliveries that resulted in injury were excluded from the control cases; those cases with no recorded shoulder dystocia were excluded from the cases. Matching was for birth weight (+/-250 g), parity, and diabetic status. Rates of precipitous and prolonged second stage, operative delivery, neonatal depression, and average number of shoulder dystocia maneuvers used were compared between the two groups with chi(2) test, Fisher exact test, and the Student t test; a probability value of <.05 was considered significant. RESULTS: There were 80 matched patients, of which 26 patients were nulliparous and 11 patients were diabetic. Mothers of the uninjured group were younger than those of the injured group (23.7+/-6.2 years vs 27.4+/-5.1 years, P<.001). The injured group had a significantly higher rate of 5-minute Apgar scores of <7 (13.9% vs 3.8%, P=.04). Differences in maternal weight, body mass index, height, race, gestational age, average number of maneuvers, head-to-body delivery interval, operative delivery rate, prolonged second stage rate, precipitous second stage rate, and sex were not significant between groups. The rates of precipitous second stage for both groups (28.0% injured and 35.0% uninjured) were more than triple the rates of prolonged second stage (9.5% injured and 11.3% uninjured). CONCLUSION: No characteristic of second-stage of labor predicts permanent brachial plexus injury. Precipitous second stage is the most prevalent labor abnormality that is associated with shoulder dystocia. PMID- 14526303 TI - Patterns of abnormal pudendal nerve function that are associated with postpartum fecal incontinence. AB - OBJECTIVE: The purpose of this study was to assess patterns of abnormal pudendal nerve function in women who complain of postpartum fecal incontinence. STUDY DESIGN: During a 12-month period, a cohort of 83 women underwent neurophysiologic assessment as part of an evaluation of fecal incontinence after vaginal delivery. Pudendal nerve assessment consisted of the measurement of the clitoral-anal reflex and quantitative electromyography of the external anal sphincter. Endoanal ultrasound examination and anal manometry were also performed in each patient. RESULTS: Thirty of 83 women (38%) with fecal incontinence were found to have abnormal neurophysiologic condition, among whom four identifiable patterns of abnormality emerged. Five women (17%) had evidence of pudendal nerve demylenation with a prolonged sensory threshold of the clitoral-anal reflex (>5.2 mA), although electromyography studies were normal. Eight women (27%) had abnormal electromyography results that were consistent with axonal neuropathy with or without reinervation, in whom the clitoral-anal reflex was normal. Thirteen women (43%) demonstrated a mixed demyelinating and axonal pudendal neuropathy, with evidence of reinervation. Four women (13%) had abnormal patterns of neurophysiologic condition that was not attributable directly to past obstetric trauma but to coincident medical problems. CONCLUSION: Four abnormal patterns of pudendal nerve function may be identified, three of which (demyelinating, axonal, and mixed demyelinating/axonal) can be attributed to specific past obstetric events, although a fourth radicular pattern is due to coincident medical or orthopedic problems. Assessment of pudendal nerve function is important in women with postpartum fecal incontinence because particular patterns of abnormality correlate with different symptoms and can influence treatment options. PMID- 14526304 TI - Clinical significance of the umbilical cord twist. AB - OBJECTIVE: The objective of this study was to determine the clinical significance of the umbilical cord twist direction. STUDY DESIGN: Two hundred singleton third trimester placentas with a right umbilical cord twist and 200 placentas with a left umbilical cord twist, which was determined by pathologic examination, were included. Maternal and neonatal outcomes were compared with the use of Fisher's exact and Mann Whitney U tests; a probability value of <.05 considered statistically significant. RESULTS: Placenta previa was more common in patients with a right umbilical cord twist compared with a left umbilical cord twist (6.0% vs 1.5%; P<.05). There was a trend towards an increased incidence of single umbilical artery in patients with a right umbilical cord twist (2.5% vs 0%; P=.06). The incidence of fetal demise, intrauterine growth restriction, chromosomal abnormalities, congenital anomalies, preterm delivery, infant gender, birth weight, maternal age, and parity were similar between the 2 groups. CONCLUSION: Placenta previa is associated with a right umbilical cord twist. PMID- 14526305 TI - Factors associated with adverse perinatal outcome in the Term Breech Trial. AB - BACKGROUND: In the Term Breech Trial, the risk of adverse perinatal outcome was lower with planned cesarean section versus planned vaginal birth. We undertook secondary analyses to determine factors associated with adverse perinatal outcome. STUDY DESIGN: By using multiple logistic regression analyses, we determined the effect of prelabor cesarean section, cesarean section during early labor, cesarean section during active labor versus vaginal birth, and other factors, on adverse perinatal outcome. For 1384 fetuses delivered after labor, we determined the effect of variables associated with labor on adverse perinatal outcome. RESULTS: The risk of adverse perinatal outcome was lowest with prelabor cesarean section (odds ratio [OR]=0.13) and highest with vaginal birth. For those delivered after labor, labor augmentation (P=.007), birth weight less than 2.8 kg (P=.003), and longer time between pushing and delivery (P<.001) increased the risk, whereas the presence of an experienced clinician at delivery (P=.004) reduced the risk of adverse perinatal outcome. CONCLUSION: Breech infants at term are best delivered by prelabor cesarean section. PMID- 14526306 TI - Meconium-stained amniotic fluid is associated with puerperal infections. AB - OBJECTIVE: The purpose of this study was to determine whether meconium-stained amniotic fluid is associated with puerperal infection and whether the quality of the meconium is further associated with this risk. STUDY DESIGN: We designed a retrospective cohort study of all deliveries beyond 37 weeks gestational age from 1992 to 2002 at a single community hospital. Data were collected on rates of chorioamnionitis, endomyometritis, quality of amniotic fluid, and length of labor and analyzed with bivariate and multivariate analyses. RESULTS: We found that, among the 43,200 women who were delivered at term, 18.9% of the women had meconium staining (8.8% thin, 5.5% moderate, 4.6% thick). Compared with deliveries with clear amniotic fluid, those with meconium-stained amniotic fluid had higher rates of chorioamnionitis (2.3% vs 4.1%, P<.001) and endomyometritis (1.0% vs 1.7%, P<.001). Further, the severity of meconium staining was associated with increased rates of infection. CONCLUSION: We found that the presence and severity of meconium-stained amniotic fluid is associated with puerperal infection even when being controlled for confounders. PMID- 14526307 TI - The relationship between amniotic fluid index and successful external cephalic version: a 14-year experience. AB - OBJECTIVE: This study was undertaken to assess the association between amniotic fluid index (AFI), success of external cephalic version (ECV), and obstetric outcome in patients undergoing ECV. STUDY DESIGN: A prospective observational study was performed that included all patients who had a trial of ECV from 1987 to 2000 in our center. Rates of success, fetal heart decelerations during trial at ECV, and cesarean deliveries were calculated. Groups were divided by an AFI performed immediately before ECV: AFI less than 10 cm, 10 to 15 cm, and more than 15 cm. RESULTS: In our group of 1361 patients undergoing ECV, the rate of success was related to the AFI in both parous women (49.1%, 63.5% and 72.1% [P<.001] for each AFI group, respectively) and nulliparous women (36.5%, 43.6%, and 57.3% [P<.001]). The rate of cesarean section delivery was related to AFI in nulliparous but not multiparous patients (P<.001). The rate of prolonged fetal heart rate decelerations was not significantly related to the AFI. CONCLUSION: The rate of successful ECV and cesarean section deliveries is related to the amniotic fluid volume. This information may be used to consent patient before a trial of ECV. PMID- 14526308 TI - Isolated low second-trimester maternal serum beta-human chorionic gonadotropin is not associated with adverse pregnancy outcome. AB - OBJECTIVE: We investigated whether low human chorionic gonadotropin from maternal serum screening is associated with adverse pregnancy outcome. STUDY DESIGN: Women between 15 and 20 completed weeks of gestation who had a maternal serum screen performed from June 1999 to November 2001 were studied. Cases included women with human chorionic gonadotropin values of 0.5 and <2.0 multiples of the median, and estriol values of >0.6 and <2.0 multiples of the median. Control subjects were selected randomly from the population of women with normal values for all three analytes. RESULTS: There were 146 case subjects and 292 control subjects. There was no increased risk in the study group compared with the control subjects for preterm delivery, intrauterine fetal death, low birth weight, abruptio placentae, preeclampsia, or preterm premature rupture of membranes. CONCLUSION: An isolated low human chorionic gonadotropin level from second-trimester maternal serum screening is not associated with adverse pregnancy outcome. PMID- 14526309 TI - Is adenovirus a fetal pathogen? AB - OBJECTIVES: The purpose of this study was to test the relationship between adenovirus genetic material in the amniotic fluid and adverse pregnancy outcome. STUDY DESIGN: This was a prospective, observational study of women who were referred in the second trimester of gestation for either genetic amniocentesis or evaluation of fetal malformation. A 2-mL aliquot of amniotic fluid was subjected to multiplex polymerase chain reaction for a panel of viruses that included adenovirus and human genome controls. Fetuses with an abnormal karyotype were excluded from analysis. RESULTS: The prevalence of adenovirus was similar in normal (39/652) and anomalous fetuses (23/364; chi(2) test, P=.376). There was significant seasonal variation in the prevalence in both normal and anomalous fetuses (chi(2) exact test, P<.001), but no significant difference between groups. The monthly proportion of patients who underwent amniocentesis remained constant throughout the year (mean, 8.3%; chi(2) test, P=.67). Central nervous system anomalies and echogenic liver foci were significantly more common among fetuses with positive amniotic fluid polymerase chain reaction results for adenovirus (P<.005, respectively). CONCLUSION: Adenovirus is found in a similar prevalence and seasonal variation in sonographically normal and abnormal pregnancies. Although a specific fetal presentation was not identified, echogenic liver lesions with or without hydrops and neural tube defects were significantly more common in the presence of adenovirus. The significance of these findings deserves further study. PMID- 14526310 TI - The epidemiologic incidence of congenital gastroschisis in Western Australia. AB - OBJECTIVE: The purpose of this study was to review the population incidence of congenital gastroschisis in Western Australia over the past 2 decades. STUDY DESIGN: A population-based incidence study of congenital gastroschisis from 1980 to 2001. Maternal and perinatal outcome data were collected to ascertain incidence, treatment, and outcome trends. RESULTS: One hundred twenty-two cases of gastroschisis were identified. The median maternal age was 23 years (range, 16 35 years). Women aged <20 years were at a 7.82 increased risk (95% CI, 4.34 14.08); women aged 20 to 24 years were at a 3.24 increased risk (95% CI, 1.88 5.61) for fetal gastroschisis compared with women aged >or=25 years. An incidence analysis over time indicated a significant increase of gastroschisis cases from 1 of 10,000 births during the period 1980-1990 to the current rate of 2.4 of 10,000 births (P<.001). The perinatal mortality rate was 12.7% (95% CI, 8.7-16.7) with a 9.8% stillbirth rate (95% CI, 6.3-13.3). CONCLUSION: There has been a sustained increase in the birth incidence of gastroschisis over the past decade, particularly in teenage women. A significant fetal death rate in the third trimester is observed. PMID- 14526311 TI - Prenatal cystic fibrosis screening in Mexican Americans: an economic analysis. AB - OBJECTIVE: We evaluated the cost benefit of cystic fibrosis screening in Mexican American gravid women. STUDY DESIGN: With the use of decisions analysis techniques, a cost-benefit analysis was performed. Baseline assumptions were based on published references. Sensitivity analyses were performed. RESULTS: Under the baseline assumptions, screening was not cost beneficial. Threshold analysis showed that, if the test was priced under 53.00 dollars, screening became cost beneficial. Sensitivity analysis demonstrated that lower acceptance rates of amniocentesis or termination made the screening strategy less attractive. If the test sensitivity was raised to 90%, which required testing of >60 mutations, the cost of screening would need to be <100.00 dollars for the program to be cost beneficial. CONCLUSION: Cystic fibrosis screening is not cost beneficial in Mexican American women over a wide range of assumptions. This is principally due to the poor sensitivity of the test in this population. Cultural factors, such as lower acceptance of amniocentesis and pregnancy termination of affected fetuses, further lower the cost-benefit ratio of screening. PMID- 14526313 TI - Neonatal outcome after exposure to indomethacin in utero: a retrospective case cohort study. AB - OBJECTIVE: This study was undertaken to determine the clinical outcome for neonates who were exposed to indomethacin during gestation. STUDY DESIGN: We identified 124 infants with in utero exposure to indomethacin and matched them to 124 infants whose mothers did not receive indomethacin. The two groups were matched for gestational age at birth, sex, and exposure to antenatal betamethasone. Sixty-two of the indomethacin-exposed infants were born within 48 hours of last exposure. These infants were also compared with their matched controls. RESULTS: There were no significant differences between the indomethacin exposed infants and control infants in birth weight, Apgar scores, frequency of cesarean section deliveries, and multiple gestation. The incidence of respiratory distress syndrome, need for surfactant treatment, patent ductus arteriosus, necrotizing enterocolitis, and intraventricular hemorrhage was similar between the indomethacin-exposed group and the control group. Indomethacin-exposed infants who were born within 48 hours of last exposure had similar incidence of respiratory distress syndrome but greater need for surfactant treatment (P=.02) compared with controls. All other complication rates were similar. CONCLUSION: Indomethacin exposure in our study was not associated with increased neonatal complications for infants delivered within or beyond 48 hours of last exposure. PMID- 14526312 TI - Unexplained elevated maternal serum alpha-fetoprotein and/or human chorionic gonadotropin and the risk of adverse outcomes. AB - OBJECTIVE: The study was undertaken to determine the risks of adverse obstetric outcomes in pregnant women with unexplained elevations of maternal serum alpha fetoprotein (MSAFP) and/or human chorionic gonadotropin (hCG) and to determine whether these risks vary by prepregnancy risk status. STUDY DESIGN: All women who underwent double-marker screening (MSAFP+hCG) between 1994 and 2000 and were delivered of an infant in Nova Scotia, Canada, during this period were identified from a hospital serum screening database and a provincial perinatal database. Patients with inaccurate dating, major structural anomalies, or chromosomal abnormalities were excluded. The primary outcomes studied were preeclampsia, abruptio placentae, fetal growth restriction, fetal death, and preterm birth. Women with medical or previous obstetric complications were designated high risk. Logistic regression, controlling for confounding factors, was used to estimate the relative risks (RRs) and 95% CI for elevated levels of MSAFP and/or hCG and each of the outcomes. RESULTS: Among the 14,374 women who met the study criteria, 5,789 were designated high risk. Except for abruptio placentae, unexplained elevated MSAFP or elevated hCG levels were independently associated with all the outcomes in both high- and low-risk women. Elevated screening values were associated with increased risk of abruptio placentae among low-risk women only. Particularly large RRs were seen for fetal death in both high- and low-risk women (RR=4.9, 95% CI 2.7-8.7 for elevated MSAFP or hCG in high- and low-risk women combined). CONCLUSION: Unexplained elevated levels of MSAFP and/or hCG are associated with an increased risk of most pregnancy complications. Increased antenatal surveillance of these patients is important regardless of prepregnancy risk status. PMID- 14526314 TI - Differential expression of c-fos in a mouse model of fetal alcohol syndrome. AB - OBJECTIVE: Fetal alcohol syndrome (FAS) results in stillbirth, fetal growth restriction, and mental retardation with injury attributed to oxidative stress. Our objective was to identify signal transduction pathways expressed in a model of FAS and to quantify expression of c-fos, a gene in the stress signal pathway. STUDY DESIGN: Timed, pregnant C57Bl6/J mice were injected on E8 with saline solution or alcohol. RNA was extracted from decidua and embryo 6 and 24 hours later. Microarray analysis was used to screen gene pathways. Differential gene expression was confirmed using real-time polymerase chain reaction with results presented as the ratio of c-fos concentration to that of glyceraldehyde-3 phosphate dehydrogenase (GAPDH). RESULTS: Differential gene expression between alcohol and control was noted for stress signal pathway genes including c-fos. Real-time polymerase chain reaction demonstrated that c-fos messenger RNA expression was greater in the alcohol than control decidua at 6 hours after injection (P<.01). This effect persisted at 24 hours (P<.01). There was no difference in c-fos expression in embryos whose mothers received alcohol versus control after 6 hours (P=.12) or 24 hours (P=.89). CONCLUSION: Alcohol administration during pregnancy results in differential gene expression in the stress signal pathway, particularly in c-fos. C-fos expression in the decidua increases from 6 to 24 hours after alcohol injection, but does not change in the embryo, which may contribute to alcohol-induced damage in FAS. PMID- 14526315 TI - The role of activity-dependent neuroprotective protein in a mouse model of fetal alcohol syndrome. AB - OBJECTIVE: Fetal alcohol syndrome (FAS) is the most common nongenetic cause of mental retardation. Peptides NAPVSIPQ (NAP) and SALLRSIPA (SAL), related to activity-dependent neuroprotective protein (ADNP), prevent alcohol-induced damage in a mouse model of FAS. Our objective was to characterize ADNP in this model to relate this protein to the mechanisms of damage and peptide neuroprotection. STUDY DESIGN: Timed, pregnant C57Bl6/J mice were treated on day 8. Groups were control, alcohol, peptide pretreatment, or peptide alone. Embryo and decidua were harvested at 6 and 24 hours and 10 days. To evaluate ADNP expression, real-time polymerase chain reaction was performed with results presented as the ratio of ADNP-to-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) concentration. Analysis of variance was performed for overall comparisons with P<.05 considered significant. RESULTS: At 6 hours, there was no difference in ADNP between alcohol exposed embryos compared with control embryos. At 24 hours, there was an increase in ADNP in alcohol-exposed embryos compared with controls (P<.001); these findings persisted at 10 days (P<.001). In the decidua at 6 hours, there was no difference between alcohol and control. At 24 hours, there was greater ADNP in alcohol-exposed decidua compared with controls (P<.001), which did not persist at 10 days (P=.97). Peptide pretreatment did not prevent the alcohol-induced increase in ADNP in embryo or decidua. CONCLUSION: Alcohol increased embryonic and decidual ADNP expression at 24 hours and it persisted in the embryo for 10 days. Because ADNP is a known neuroprotectant, these findings suggest that it may be released as a protective mechanism in FAS. Changes in the embryo were persistent suggesting that the embryo is more vulnerable to alcohol-induced damage than the mother. PMID- 14526316 TI - Prolonged latency after preterm premature rupture of membranes: an evaluation of histologic condition and intracranial ultrasonic abnormality in the neonate born at <28 weeks of gestation. AB - OBJECTIVE: The purpose of this study was to evaluate whether infants who were delivered at <28 weeks of gestation after prolonged latency in pregnancies that were complicated by preterm premature rupture of membranes are at increased risk of histologic chorioamnionitis and intracranial ultrasound abnormalities. STUDY DESIGN: A retrospective cohort analysis of 430 singleton infants born at <28 weeks of gestation in five hospitals (January 1991 through December 1993) with at least one of three protocol cranial scans read by a consensus committee and with placental pathologic evidence. Outcome variables were placental (histologic chorioamnionitis, fetal vasculitis) and neonatal (intraventricular hemorrhage, echolucencies, ventriculomegaly). Latency was divided into five intervals, and outcomes in the longer four intervals were compared with those in infants who were delivered at <1 hour after membrane rupture. Each outcome-latency relationship was evaluated in a logistic model that was controlled for confounders. RESULTS: Odds ratios and CIs for each latency interval that was controlled for confounders that included gestational age, maternal race, antenatal steroid and antibiotic administration, and delivery mode show a statistically significant increase in the risk of histologic chorioamnionitis and fetal vasculitis. Models for intraventricular hemorrhage, ventriculomegaly, and echolucencies failed to demonstrate significant differences with increasing latency. CONCLUSIONS: Ascending transcervical infection after preterm premature rupture of membranes is documented by the increasing odds ratios of placental inflammation. The odds of ultrasonically detectable brain abnormalities, however, did not increase with increasing latency. PMID- 14526317 TI - Duration of antibiotic therapy after preterm premature rupture of fetal membranes. AB - OBJECTIVE: This study was undertaken to compare the efficacy of 3 days versus 7 days of ampicillin in prolonging gestation for at least 7 days in women with preterm premature rupture of membranes (PPROM). STUDY DESIGN: We performed a randomized clinical trial comparing 3 days of ampicillin with 7 days ampicillin in patients with PPROM. Our primary outcome was the prolongation of pregnancy for at least 7 days. Secondary outcomes included rates of chorioamnionitis, postpartum endometritis, and neonatal morbidity and mortality. RESULTS: Forty eight patients were randomly selected. There was no statistically significant difference in the ability to achieve a 7-day latency (relative risk 0.83, 95% CI 0.51-1.38). In addition, there was no statistically significant difference in the rates of chorioamnionitis, endometritis, and our composite neonatal morbidity. CONCLUSION: In patients with PPROM, length of antibiotic therapy does not change the rate of a 7-day latency or affect the rate of chorioamnionitis, postpartum endometritis, or neonatal morbidity. PMID- 14526318 TI - Umbilical cord pH and base excess values in relation to neonatal morbidity for infants delivered preterm. AB - OBJECTIVE: Our purpose was to determine the relationship/predictive value of umbilical cord pH and base excess (BE) values to adverse neonatal outcomes for preterm infants. STUDY DESIGN: A tertiary center perinatal/neonatal database was used to obtain umbilical cord pH and BE values, adverse neonatal outcomes, and patient demographics for preterm (PT, 32-36 weeks gestational age) and very preterm (VPT, 25-32 weeks gestational age) singleton live-born infants delivered between November 1995 and March 2002. RESULTS: PT (n=1807) and VPT (n=603) groups demonstrated a significant inverse curvilinear relationship of umbilical cord pH and BE values to Apgar score <7 at 5 minutes, respiratory distress syndrome, assisted ventilation, and intraventricular hemorrhage/periventricular leukomalacia. Receiver operating characteristic area under the curve values ranged from 0.69 to 0.86 (PT) and 0.70 to 0.87 (VPT). There was little difference between umbilical vein, umbilical artery, pH, or BE in predictive value. CONCLUSION: Umbilical cord pH and BE are related to subsequent adverse outcome events for infants delivered preterm. Worsening acidosis is associated with progressively greater increases in these outcomes with no discriminatory value within or between umbilical artery and umbilical vein pH and BE. PMID- 14526319 TI - Neutrophil granule products: can they identify subclinical chorioamnionitis in patients with preterm premature rupture of membranes? AB - OBJECTIVE: Our purpose was to determine whether maternal plasma levels of neutrophil granule products are elevated in patients with chorioamnionitis after preterm premature rupture of membranes (PROM). STUDY DESIGN: Fifty-two patients between 24 and 34 weeks' gestation with preterm PROM were included. Plasma samples for defensins and lactoferrin were collected throughout latency. Fifty two control subjects between 26 and 30 weeks' gestation were recruited for baseline levels. RESULTS: Mean control defensin levels were compared with mean defensin levels on admission (668 ng/mL vs 5665 ng/mL, P<.01). Mean defensin levels on admission in patients without chorioamnionitis were compared with those of patients in whom histologic chorioamnionitis developed (520 ng/mL vs 9163 ng/mL, P<.01). The same relationships were not demonstrated for lactoferrin. With use a defensin value of 1500 ng/mL on admission, the sensitivity is 76% and specificity is 94% in predicting histologic chorioamnionitis. CONCLUSIONS: Maternal plasma levels of defensins are markers of histologic chorioamnionitis in patients after preterm PROM. PMID- 14526320 TI - Antenatal factors associated with significant birth weight discordancy in twin gestations. AB - OBJECTIVE: The purpose of this study was to evaluate factors that are associated with significant birth weight discordancy. STUDY DESIGN: As a part of an ongoing collaborative study of twins, maternal and fetal data were obtained from the medical records of twin gestations at eight medical centers. The study population was divided into groups by difference in birth weight discordancy (>or=20%, >or=25%, and >or=30%) RESULTS: Severe birth weight discordancy was associated with fetal growth deceleration by 20 to 28 weeks (adjusted odds ratio, 4.90; 95% CI, 3.15-7.64) and between 28 weeks to birth (adjusted odds ratio, 3.48; 95% CI, 1.72-7.06). Antenatal bleeding (adjusted odds ratio, 1.86; 95% CI, 1.08-3.21), preeclampsia (adjusted odds ratio, 1.70, 95% CI, 1.21-2.41), and monochorionicity (adjusted odds ratio, 2.35, 95% CI, 11.71-3.23) were also associated with birth weight discordancy. CONCLUSION: These data demonstrate the importance of the early diagnosis of placental chorionicity, because monochorionicity is associated with a 2-fold increase in birth weight discordancy in twin gestations. PMID- 14526322 TI - The impact of prenatal care on preterm births among twin gestations in the United States, 1989-2000. AB - OBJECTIVE: The purpose of this study was to determine the association between prenatal care and preterm births among twin gestations in the presence and absence of high-risk pregnancy conditions. STUDY DESIGN: Twin birth data in the United States were used to determine the association between preterm birth and prenatal care with the use of logistic regression. RESULTS: Of the 779,387 twin births, 54.7% twin births were delivered preterm. The rate was higher among black women than among white women in the presence (57.0% vs 51.2%, respectively) and absence (70.3% vs 61.6%, respectively) of prenatal care. The absence of prenatal care increased the relative risk for preterm birth by 1.24-fold among black women and by 1.22-fold among white women. Lack of prenatal care was associated with increased preterm birth rates in the presence of most high-risk conditions. CONCLUSION: Prenatal care is associated with fewer twin preterm births in the presence and absence of high-risk conditions. Increased prenatal care participation may help decrease preterm birth rates and also narrow the black white twin preterm birth disparity. PMID- 14526323 TI - Tertiary care improves the chance for vaginal delivery in women with preeclampsia. AB - OBJECTIVE: The purpose of this study was to determine whether the level of hospital care affects cesarean delivery rates for women with preeclampsia. STUDY DESIGN: We conducted a population-based cohort study using Missouri birth certificate data for 1993 through 1999. Logistic regression was used to analyze data from 13,646 nulliparous women with preeclampsia who were delivered of singleton live births. RESULTS: After adjustment was made for gestational age and birth weight, the data showed that women with preeclampsia at primary and secondary hospitals were more likely to be delivered by cesarean delivery (odds ratio, 1.37; 95% CI, 1.24,1.51; and odds ratio, 1.16; 95% CI, 1.07,1.26, respectively) than at tertiary hospitals. For women who were delivered at >or=37 weeks of gestation, cesarean delivery rates were 38.0%, 33.7%, and 30.0% for primary, secondary, and tertiary hospitals, respectively. Dysfunctional labor, cephalopelvic disproportion, and fetal distress were more commonly noted at primary and secondary hospitals (P<.001). CONCLUSION: Levels of expertise and staffing at tertiary hospitals may allow greater attempts and success with vaginal delivery among women with preeclampsia compared with primary or secondary hospitals. PMID- 14526324 TI - Maternal benefit of high-dose intravenous corticosteroid therapy for HELLP syndrome. AB - OBJECTIVE: We compared maternal outcomes for patients with HELLP (hemolysis, elevated liver enzymes, and low platelet count) syndrome treated with or without high-dose corticosteroids to ameliorate maternal disease. STUDY DESIGN: An analysis of data for patients with HELLP syndrome (platelets, or=600 IU/L; aspartate aminotransferase and/or alanine aminotransferase level, >or=70 IU/L) who were treated during the 7-year epochs before and after the clinical trials in 1992 and 1993 demonstrated maternal benefit with high-dose dexamethasone. RESULTS: Corticosteroid use increased from 16% (39/246 patients) for fetal indication from 1985 to 1991 to 90% (205/228 patients) for maternal-fetal indications from 1994 to 2000. Significantly reduced composite maternal disease from 1994 to 2000 was evidenced by improvements in laboratory parameters, disease progression to class 1 HELLP syndrome, the degree of hypertension, the need for antihypertensive therapy, the use of transfusion, and the presence of maternal morbidity (P<.05). Indices of postpartum recovery also were shortened significantly (P<.001). CONCLUSION: Routine early initiation of high-dose intravenous corticosteroids for patients with HELLP syndrome significantly lessened maternal disease, reduced maternal morbidity, and expedited recovery. PMID- 14526325 TI - The effects of a cyclo-oxygenase II inhibitor on placental artery production of thromboxane and prostacyclin. AB - OBJECTIVE: The study was undertaken to determine the effects of a cyclo-oxygenase II inhibitor on fetoplacental artery production of prostacyclin and thromboxane A(2). STUDY DESIGN: Eight placentas were obtained from normal parturients at delivery and four chorionic plate arteries were dissected from each placenta. Arteries were incubated in media alone, media plus angiotensin II (1x10(-10) mol), media plus rofecoxib (300 ng/mL), or media plus angiotensin II and rofecoxib. Serial samples were assayed for metabolites of thromboxane B(2) and prostacyclin by enzyme-linked immunosorbent assay. Results were compared by analysis of variance, and P<.05 was considered significant. RESULTS: At 24 hours, 6-keto-prostaglandin F(1alpha) levels in the rofecoxib group (1.74+/-1.39 ng/mg tissue, P<.01) and the rofecoxib plus angiotensin II group (2.15+/-1.85 ng/mg tissue, P<.01) were significantly lower than levels in the control group (4.25+/ 2.03 ng/mg tissue). Thromboxane B(2) levels were lower in the angiotensin II group (0.65+/-0.33 ng/mg tissue) than the control group (1.22+/-0.70 ng/mg tissue, P<.05). CONCLUSION: Cyclo-oxygenase II inhibition decreases the production of prostacyclin in fetoplacental arteries and alters the normal ratio of thromboxane A(2) to prostacyclin. PMID- 14526326 TI - Fetoplacental vascular tone is modified by magnesium sulfate in the preeclamptic ex vivo human placental cotyledon. AB - OBJECTIVE: The purpose of this study was to evaluate fetoplacental vascular tone and response to a vasoconstrictor in placentas of preeclamptic and normotensive pregnancies with and without the presence of magnesium sulfate. STUDY DESIGN: Two cotyledons from each placenta were selected from preeclamptic (n=8) and normotensive (n=7) pregnancies. In one cotyledon from each pair, the maternal circuit was perfused with magnesium sulfate. The fetal arteries were injected sequentially with angiotensin II (10(-10)mol and 10(-11.5) mol). Perfusion pressures and response to angiotensin II were compared, with regard to preeclampsia and exposure to magnesium sulfate. RESULTS: Perfusion pressure was higher in preeclamptic placentas, compared with normotensive placentas (30.4 mm Hg vs 24.4 mm Hg, P=.02). There was a decrease in perfusion pressure with exposure to magnesium sulfate in preeclamptic placentas (22.5 mm Hg, P<.01), but not in normotensive placentas. Fetoplacental vascular response to angiotensin II was not affected by preeclampsia or magnesium sulfate. CONCLUSION: In placentas from preeclamptic pregnancies there is increased fetoplacental perfusion pressure, which decreases with exposure to sulfate. PMID- 14526327 TI - Differential expression of TcR-CD3 zeta as evidence for altered immunoregulation in preeclamptic versus normotensive women. AB - OBJECTIVE: The study was undertaken to exhibit and quantify the difference in modulation of CD3-zeta protein (an integral component of the T-cell receptor) in preeclamptic and normotensive women. STUDY DESIGN: Serum was collected from 10 preeclamptic and 10 normotensive women at >or=37 weeks' gestation on admission. Jurkat E-61 cells were incubated with the sera (20% volume to volume) and analyzed with Western immunoblot using mouse monoclonal CD3-zeta antibody. Enhanced chemiluminescence and densitometry were used to qualitatively measure zeta expression of the cells. A de novo flow cytometry assay was developed to quantify the difference in CD3-zeta expression of these cells. Comparisons were performed by t test (P<.05 was significant). RESULTS: Preeclamptic patient sera produced a 2.4-fold increase in CD3-zeta expression than normotensive patients on Western blot (P<.01). Flow cytometry showed that preeclamptic sera had a 1.4-fold higher expression of CD3-zeta compared with normotensive patients (P<.0003). CONCLUSION: TcR/CD3-zeta expression is normally suppressed in pregnancy. Loss of this suppression occurs in preeclamptic patients, implying increased T-cell function. PMID- 14526328 TI - A prospective comparison of total protein/creatinine ratio versus 24-hour urine protein in women with suspected preeclampsia. AB - OBJECTIVE: The purpose of this study was to determine the value of the protein/creatinine ratio in prediction of 24-hour urine total protein among women with suspected preeclampsia. STUDY DESIGN: Women who were evaluated for suspected preeclampsia at >or=24 weeks of gestation were studied prospectively if there was no concurrent diagnosis of chronic hypertension, diabetes mellitus, or preexisting renal disease. A protein/creatinine ratio was obtained, which was followed by the initiation of a 24-hour urine evaluation. Positive and negative predictive values and sensitivity and specificity of the protein/creatinine ratio for significant (>or=300 mg) and severe proteinuria (>or=5000 mg) that were based on 24-hour urine total protein were calculated. RESULTS: A total of 220 women were evaluated; 43.2% of the women were black, and 80% of the women had government insurance. Mean maternal and gestational ages were 26.1 years and 36.5 weeks, respectively. Significant and severe proteinuria on 24-hour urine evaluation were identified in 76.4% and 8.2% of cases, respectively. Regression analysis of protein/creatinine ratio and 24-hour urine total protein level showed a poor correlation (r(2)=0.41). Receiver operator characteristic analysis revealed an area under the curve of 0.80, but the shoulder value of 390 mg/g carried a high false-negative rate (45.2%). With a more conservative cutoff value, a protein/creatinine ratio of >or=300 mg/g had a poor negative predictive value (47.5%), a specificity for significant proteinuria (55.8%), with a positive predictive value of 85.5%, and a sensitivity of 81%. For severe proteinuria, a protein/creatinine ratio of >or=5000 mg/g had a poor positive predictive value (61.9%) and sensitivity (72.2%), with a negative predictive value of 97.5%, and a specificity of 96.0%. CONCLUSION: Protein/creatinine ratio does not exclude adequately the presence of significant proteinuria or predict severe proteinuria and should not be used as an alternative to 24-hour total protein evaluation. PMID- 14526329 TI - Pregnancy outcome in women treated with doxorubicin for childhood cancer. AB - OBJECTIVE: Pregnancy outcome and the effect of pregnancy on cardiac function were assessed in women cured of childhood cancer. STUDY DESIGN: Forty women who received doxorubicin as part of a chemotherapy protocol for a neoplastic disorder in childhood were followed up at the same center during pregnancy and after delivery. RESULTS: Thirty-seven women (72 pregnancies) completed follow-up. Pregnancy outcome was favorable in the 29 women with fractional shortening values of >or=30% before pregnancy, and their myocardial function was sustained. In 8 women with fractional shortening of <30% before pregnancy, pregnancy outcome was less favorable; a 19% decrease in fractional shortening was observed after pregnancy, and this finding was not significant (P=.08). CONCLUSION: Pregnancy outcome in women who received doxorubicin for malignancy in childhood is generally favorable. However, those with baseline left ventricular dysfunction should be considered at increased risk for worse pregnancy outcome and further deterioration in myocardial function. PMID- 14526330 TI - Evaluation of a clinical skills orientation program for residents. AB - OBJECTIVE: Our purpose was to implement and evaluate an orientation program for residents, focusing on outpatient clinical skills. STUDY DESIGN: Eleven of 12 residents participated in a clinical skills orientation program immediately preceding the academic year. The skill stations included evaluation of abnormal uterine bleeding, intrauterine device insertion, basic infertility evaluation, endometrial and vulvar biopsies, pelvic organ prolapse quantification examination, hysterosalpingography and office hysteroscopy, ultrasound scanning, labor and delivery triage, and clinic administrative responsibilities. Before test, after test, and anonymous resident evaluations were used to evaluate the program. RESULTS: First-year residents demonstrated a statistically significant increase in posttest scores compared to pretest scores (42.5% vs 71.3%, P=.003). Only first-year resident posttest scores for the labor and delivery triage and basic infertility evaluation stations demonstrated statistically significant increases over pretest scores (14.3% vs 46.4%, P=.009; and 41.7% vs 83.3%, P=.049, respectively). Sixty-four percent of the residents rated the program as "very helpful." Most residents felt that the program was well organized and that the facilities were conducive to learning; all of the participants recommended an annual clinical orientation program. CONCLUSION: A clinical skills orientation program was well received and strongly desired by residents. First-year residents appeared to benefit the most from this orientation. PMID- 14526331 TI - The infectious origins of stillbirth. AB - OBJECTIVE: Our objective was to determine the relationship between various types of perinatal infections and stillbirths. STUDY DESIGN: By use of various textbooks on perinatal infections, multiple MEDLINE searches, and the reference list of all appropriate manuscripts, the appropriate English language literature was reviewed to define the relationship between various perinatal infections and stillbirths. RESULTS: Infection may cause stillbirth by a number of mechanisms, including direct infection, placental damage, and severe maternal illness. A large variety of organisms have been associated with stillbirth, including many bacteria, viruses, and protozoa. In developed countries, between 10% and 25% of stillbirths may be caused by an infection, whereas in developing countries, which often have much higher stillbirth rates, the contribution of infection is much greater. Ascending bacterial infection, both before and after membrane rupture, with organisms such as Escherichia coli, group B streptococci, and Ureaplasma urealyticum is usually the most common infectious cause of stillbirth. However, in areas where syphilis is very prevalent, up to half of all stillbirths may be caused by this infection alone. Malaria may be an important cause of stillbirth in women infected for the first time in pregnancy. The two most important viral causes of stillbirth are parvovirus and Coxsackie virus, although a number of other viral infections appear to be causal. Toxoplasma gondii, leptospirosis, Listeria monocytogenes, and the organisms that cause leptospirosis, Q fever, and Lyme disease have all been implicated as etiologic for stillbirth. CONCLUSION: Because infection-related stillbirth is relatively rare in developed countries, and those that do occur are caused by a wide variety of organisms, reducing this etiologic component of stillbirth much further will be difficult. However, in certain developing countries, the stillbirth rate is so high and the infection related component so great that achieving a substantial reduction in stillbirth should be possible simply by reducing maternal infections. PMID- 14526332 TI - Fetal anemia as a response to prophylactic platelet transfusion in the management of alloimmune thrombocytopenia. AB - The antenatal management of alloimmune thrombocytopenia (ATP) frequently requires fetal blood sampling and prophylactic platelet transfusion. A recent case of ATP complicated by associated red blood cell alloimmunization demonstrated severe hemolysis apparently as a result of ABO incompatibility from transfused platelets. PMID- 14526333 TI - An initial Glasgow score of 4 and Apgar scores of 9 and 9: a case report of a pregnant comatose woman. AB - A 28-year-old woman admitted to the emergency department with a Glascow Coma Scale score of 4 after a closed head injury resulting from a motor vehicle crash was found to have a positive serum beta-human chorionic gonadotropin level of 27 mIU/mL. After stabilization and treatment for her closed head injury, she remained in a comatose state for more than 240 days. At 36 to 37 weeks' gestation, she had contractions and elevations in her blood pressure. A healthy female infant was born by an operative vaginal delivery with Apgar scores of 9 and 9. PMID- 14526334 TI - Early resolution of increased nuchal translucency in a fetus with trisomy 18. AB - The normal timing for first-trimester nuchal translucency screening of aneuploidies is 10 to 14 weeks' gestation. We describe a fetus with trisomy 18 that presented at 11 weeks with increased nuchal thickness. Reevaluation at 12 and 13 weeks showed early return to normal of the increased nuchal measurement. PMID- 14526335 TI - Prenatal diagnosis of atraumatic fetal subdural hematoma. AB - We present a case of atraumatic subdural hematoma diagnosed by ultrasound at 22 weeks. A hyperechogenic round mass was identified in the posterior fossa. Further investigation with fetal brain magnetic resonance imaging confirmed the diagnosis. After medical consultation, the parents opted for pregnancy termination. The pathology report confirmed the ultrasonographic and magnetic resonance imaging diagnosis. PMID- 14526336 TI - Multiple vesical calculi and complete vaginal vault prolapse. AB - Multiple-vessel calculi in the setting of pelvic organ prolapse is rare. Long standing prolapse and bladder outlet obstruction, coupled with chronic infection are suspected to be the inciting factors in this case. To remove the calculi, an open cystolithotomy was performed at the time of an abdominal anti-incontinence procedure. PMID- 14526337 TI - Successful medical abortion of a pregnancy within a noncommunicating rudimentary uterine horn. AB - In asymptomatic patients with an early gestation located in a noncommunicating rudimentary horn, methotrexate may provide another treatment option or act as a useful adjunct to surgical intervention. We present a case of a successful pregnancy termination of a gestation located in a noncommunicating rudimentary horn with subsequent elective laparoscopic resection. PMID- 14526338 TI - A case of lupus peritonitis and cystitis after ovulation induction therapy. AB - We report a patient in whom lupus peritonitis and cystitis developed after ovulation induction therapy with human menopausal and chorionic gonadotropins followed by in vitro fertilization and embryo transfer. The lupus peritonitis and cystitis presented clinically as an acute abdomen. This disease should not be misdiagnosed as a nonspecific or infectious pelvic peritonitis, especially after oocyte retrieval. PMID- 14526339 TI - Inflammatory myoblastic tumor of the uterus and interleukin-6. AB - We report a patient with inflammatory myoblastic tumor of the uterus with constitutional symptoms. Surgical excision was curative and brought a rapid normalization in the elevated serum level of interleukin-6. We revealed an overproduction of interleukin-6 by a detection of messenger RNA by use of reverse transcriptase-polymerase chain reaction and immunohistochemistry. PMID- 14526340 TI - Bilateral hematosalpinx in a case of ectopic pregnancy: a clinical dilemma. AB - A 31-year-old woman with a positive pregnancy test and a transvaginal ultrasound scan result that was suggestive of a right tubal ectopic pregnancy underwent a laparoscopy, which showed bilateral hematosalpinx. In the presence of active bleeding and deteriorating hemodynamic status of the patient, a minilaparotomy was performed that revealed a right-sided hematosalpinx and a left-sided ectopic gestation. PMID- 14526341 TI - The treatment of Sjogren's syndrome with tibolone: a case report. AB - We report the successful treatment with tibolone of a postmenopausal woman affected by primary Sjogren's syndrome. One year after the beginning of treatment, the woman does not need artificial tears and vaginal lubricants. This is the first report of an effective pharmacologic treatment for primary Sjogren's syndrome in humans. PMID- 14526343 TI - Maternal serologic screening for toxoplasmosis. AB - Congenital toxoplasmosis is a rare, but potentially serious, problem during pregnancy. Toxoplasmosis is caused by a protozoal parasite that can be found in warm-blooded animals (including humans); dried cat feces, contaminated soil, or contaminated water; and raw or undercooked meat containing infective tissue cysts. Although cats play a role in the epidemiology of the disease, there is no statistical correlation between toxoplasmosis infection and cat ownership. Toxoplasmosis can be transmitted to the fetus in utero through transplacental transmission. Both the incidence of placental transmission and severity of congenital disease depend on gestational age at which maternal seroconversion occurs. Although transmission rates from mother to fetus tend to be low early in pregnancy, fetal disease severity is highest when the fetus is infected early in gestation. Serological tests to determine maternal seroconversion are available, but their use can pose ethical and practical dilemmas. Universal maternal screening is not currently warranted in the United States because disease prevalence is low. PMID- 14526344 TI - Ice massage for the reduction of labor pain. AB - The current study investigated the use of ice massage of the acupressure energy meridian point large intestine 4 (LI4) to reduce labor pain during contractions. LI4 is located on the medial midpoint of the first metacarpal, within 3 to 4 mm of the web of skin between the thumb and forefinger. A one-group, pretest, posttest design was chosen, which used 100-mm Visual Analog Scales (VAS) and the McGill Pain Questionnaire (MPQ) ranked numerically and verbally to measure pain levels; the pretest served as the control. Study participants were Hispanic and white Medicaid recipients who received prenatal care at a women's clinic staffed by certified nurse-midwives and obstetricians. Participants noted a pain reduction mean on the VAS of 28.22 mm on the left hand and 11.93 mm on the right hand. The postdelivery ranked MPQ dropped from number 3 (distressing) to number 2 (discomforting). The study results suggest that ice massage is a safe, noninvasive, nonpharmacological method of reducing labor pain. PMID- 14526345 TI - Is the childbirth experience improved by a birth plan? AB - From antenatal clinics in Sweden, 271 women were recruited after week 33 of pregnancy and given a questionnaire designed to assess their attitudes and feelings about the coming childbirth. Thereafter, they formulated a birth plan. The midwife in attendance at the birth was able to refer to this plan. Women who followed this program were compared with women from the same clinics who were asked to complete a questionnaire during the first postpartum week to assess their birth experience. A questionnaire at the end of pregnancy, followed by a birth plan, was not effective in improving women's experiences of childbirth. In the birth plan group, women gave significantly lower scores for the relationship to the first midwife they met during delivery, with respect to listening and paying attention to needs and desires, support, guiding, and respect. Although a birth plan did not improve the experience of childbirth in the overall group, there may be beneficial effects with regard to fear, pain, and concerns about the newborn for certain subgroups of women. PMID- 14526346 TI - Breast implants and breast cancer screening. AB - Concern about breast prostheses impairing breast cancer detection has become a priority issue. This article provides a review of the literature on the influence of implants on early detection methods of breast cancer, specifically breast self examination (BSE), clinical breast examination (CBE), and mammography. Background information on implants is presented, including indications for surgery, types of prostheses, location of placement, and associated complications. Conclusions from the literature suggest that the presence of implants may facilitate BSE and CBE, yet challenge interpretation of mammography. However, there is no evidence that these women will have a later stage diagnosis or a poorer prognosis if diagnosed with breast cancer. PMID- 14526348 TI - Conducting international research in midwifery: a workshop held at the ICM Congress, Vienna, April 2002. AB - A workshop on international research in midwifery was held at the International Confederation of Midwives (ICM) Triennial Congress in Vienna, April 2002. Thirty five participants from 12 countries took part. The participants themselves defined the agenda, and subsequent discussion addressed the following issues: international research relationships and collaboration; ethical conduct in international research in midwifery; the role of the International Confederation of Midwives in international research; and identifying topics for an international midwifery research agenda. Recommendations arising from this workshop were as follows: develop guidelines and a code of ethics for the conduct of international research in midwifery; continue to actively support research and further develop that support; support education and capacity building for research at basic and continuing education levels; and update on a regular basis the priorities identified for collaborative international studies. PMID- 14526347 TI - Accuracy of self-reported height and weight in women: an integrative review of the literature. AB - Height and weight are two of the most commonly used anthropometric measurements in clinical practice and research. Self-reported height and weight measurement is a simple, efficient, inexpensive, and non-invasive method of collecting data from large numbers of people. This integrative review of the published research examined the accuracy of self-reported height and weight measurements in women. Twenty-six studies examined the accuracy of self-reported height in 39,244 women. Twenty-one of the studies found that women overestimate height. Thirty-four studies reviewed the accuracy of self-reported weight in 57,172 women, and all 34 studies reported that women underestimated weight. Although mean variations between self-reported and measured values were small, a significant percentage of women in study groups had very large errors. Inaccurate measurements of both height and weight can cause significant inaccuracies in calculation of body mass index, which is used as a guide for identifying persons at risk for disease. These findings indicate that direct measurement of height and weight should be performed whenever possible for optimal measurements in clinical practice and clinically oriented research. PMID- 14526350 TI - Group A streptococcus and streptococcal toxic shock syndrome: a postpartum case report. AB - Streptococcal toxic shock syndrome from group A streptococcal disease is rare in the postpartum period, yet it is associated with high morbidity and mortality. Early diagnosis and treatment can lessen complications. Midwives can improve provision of postpartum care by being adequately educated about signs and symptoms of this disease, thereby enhancing their ability to recognize it and obtain the appropriate referral or collaborative medical care. A clinical presentation of postpartum streptococcal toxic shock syndrome, including provider follow-up, is presented. PMID- 14526349 TI - Translation and validation of the Breastfeeding Self-Efficacy Scale into Chinese. AB - The purpose of this methodological study was to translate the Breastfeeding Self Efficacy Scale (BSES) into Mandarin and determine the psychometric characteristics among a sample of 186 Chinese women. The BSES was translated and pilot-tested with 21 breastfeeding women. Following minor revisions to ensure content and semantic equivalence, the instrument was administered to 186 hospitalized breastfeeding women, who were then contacted again at 4 and 8 weeks postpartum to determine their infant feeding method and readminister the Edinburgh Postnatal Depression Scale. The psychometric assessment method used to validate the original BSES (English version) was replicated with the translated Chinese version. The Cronbach's alpha coefficient for the Chinese version BSES was 0.93, suggesting good reliability. Consistent with the English version, the factor analysis procedures resulted in the theorized two-factor solution and as hypothesized, the BSES was significantly correlated with the Edinburgh Postnatal Depression Scale, a theoretically related construct. Support for predictive validity was demonstrated through significant mean differences in BSES scores between breastfeeding and bottle-feeding mothers at 4 and 8 weeks postpartum. The results from this study are the first to examine the psychometric characteristics of the Chinese version BSES administered in a hospital postnatally and provide further evidence of the reliability and validity of the instrument. PMID- 14526351 TI - Resources for evidence-based practice, September/October 2003. PMID- 14526352 TI - Folic acid--what's it all about? PMID- 14526361 TI - Early-onset group B strep infection in newborns: prevention and prophylaxis Number 2, April 2003 (replaces Clinical Bulletin number 2, January 1997). PMID- 14526373 TI - From developmental disorder to heritable cancer: it's all in the BMP/TGF-beta family. AB - Transforming growth factor-beta (TGF-beta) regulates many cellular processes through complex signal-transduction pathways that have crucial roles in normal development. Disruption of these pathways can lead to a range of diseases, including cancer. Mutations in the genes that encode members of the TGF-beta pathway are involved in vascular diseases as well as gastrointestinal neoplasia. More recently, they have been implicated in Cowden syndrome, which is normally associated with mutations in the phosphatase and tensin homologue gene PTEN. Molecular studies of TGF-beta signalling are now showing why mutations in genes that encode components of this pathway result in inherited cancer and developmental diseases. PMID- 14526374 TI - Advances in Duchenne muscular dystrophy gene therapy. AB - Since the initial characterization of the genetic defect for Duchenne muscular dystrophy, much effort has been expended in attempts to develop a therapy for this devastating childhood disease. Gene therapy was the obvious answer but, initially, the dystrophin gene and its product seemed too large and complex for this approach. However, our increasing knowledge of the organization of the gene and the role of dystrophin in muscle function has indicated ways to manipulate them both. Gene therapy for Duchenne muscular dystrophy now seems to be in reach. PMID- 14526375 TI - The production of recombinant pharmaceutical proteins in plants. AB - Imagine a world in which any protein, either naturally occurring or designed by man, could be produced safely, inexpensively and in almost unlimited quantities using only simple nutrients, water and sunlight. This could one day become reality as we learn to harness the power of plants for the production of recombinant proteins on an agricultural scale. Molecular farming in plants has already proven to be a successful way of producing a range of technical proteins. The first plant-derived recombinant pharmaceutical proteins are now approaching commercial approval, and many more are expected to follow. PMID- 14526376 TI - Transgene introgression from genetically modified crops to their wild relatives. AB - Transgenes engineered into annual crops could be unintentionally introduced into the genomes of their free-living wild relatives. The fear is that these transgenes might persist in the environment and have negative ecological consequences. Are some crops or transgenic traits of more concern than others? Are there natural genetic barriers to minimize gene escape? Can the genetic transformation process be exploited to produce new barriers to gene flow? Questions abound, but luckily so do answers. PMID- 14526377 TI - The GM public debate: context and communication strategies. AB - Science communication is developing a new approach that promotes dialogue between scientists and the public. A recent example is the debate on the possible introduction of genetically modified crops into the United Kingdom. As this exercise in public engagement draws to a close, we consider the context in which this debate has taken place, and the challenges of developing such interactions between science and society. PMID- 14526378 TI - A future for transgenic livestock. AB - The techniques that are used to generate transgenic livestock are inefficient and expensive. This, coupled with the fact that most agriculturally relevant traits are complex and controlled by more than one gene, has restricted the use of transgenic technology. New methods for modifying the genome will underpin a resurgence of research using transgenic livestock. This will not only increase our understanding of basic biology in commercial species, but might also lead to the generation of animals that are more resistant to infectious disease. PMID- 14526379 TI - Intellectual property protection for plant-related inventions in Europe. AB - The economic and political issues that accompany the commercial growing of genetically modified crops, as well as the risk of transgene spread, are often top of the agenda for debate. But one important aspect is frequently overlooked- the intellectual property protection of plant-related inventions. What protection does European patent law afford to such inventions, how does it compare with the United States law and what are the consequences of the differences between them? PMID- 14526380 TI - GM crops: science, politics and communication. AB - As the public debate in Europe about genetically modified (GM) crops heats up and the trade row between the United States and the European Union over GM food escalates, what better time to examine the issues with an international group of experts (Box 1). Their views are diverse, but they all agree that we need more impartial communication, less propaganda and an effective regulatory regime that is based on a careful case-by-case consideration of GM technology. It seems that GM crops are here to stay, so let us hope that these requirements are met and that the developing nations that perhaps have the most to gain from this technology can start to reap its benefits. PMID- 14526381 TI - Antiplatelet therapy: in search of the 'magic bullet'. AB - The central importance of platelets in the development of arterial thrombosis and cardiovascular disease is well established. No other single cell type is responsible for as much morbidity and mortality as the platelet and, as a consequence, it represents a major target for therapeutic intervention. The growing awareness of the importance of platelets is reflected in the increasing number of patients receiving antiplatelet therapy, a trend that is likely to continue in the future. There are, however, significant drawbacks with existing therapies, including issues related to limited efficacy and safety. The discovery of a 'magic bullet' that selectively targets pathological thrombus formation without undermining haemostasis remains elusive, although recent progress in unravelling the molecular events regulating thrombosis has provided promising new avenues to solve this long-standing problem. PMID- 14526382 TI - Therapeutic potential of venom peptides. AB - Venomous animals have evolved a vast array of peptide toxins for prey capture and defence. These peptides are directed against a wide variety of pharmacological targets, making them an invaluable source of ligands for studying the properties of these targets in different experimental paradigms. A number of these peptides have been used in vivo for proof-of-concept studies, with several having undergone preclinical or clinical development for the treatment of pain, diabetes, multiple sclerosis and cardiovascular diseases. Here we survey the pharmacology of venom peptides and assess their therapeutic prospects. PMID- 14526384 TI - Mitochondrial toxicity of NRTI antiviral drugs: an integrated cellular perspective. AB - Highly active antiretroviral therapy (HAART) regimes based on nucleoside reverse transcriptase inhibitors (NRTIs) have revolutionized the treatment of AIDS in recent years. Although HAART can successfully suppress viral replication in the long term, it is not without significant toxicity, which can seriously compromise treatment effectiveness. A major toxicity that has been recognized for more than a decade is NRTI-related mitochondrial toxicity, which manifests as serious side effects such as hepatic failure and lactic acidosis. However, a lack of understanding of the mechanisms underlying mitochondrial toxicity has hampered efforts to develop novel drugs with better side-effect profiles. This review characterizes the pharmacological mechanisms and pathways that are involved in mitochondrial dysfunction caused by NRTIs, and suggests opportunities for future pharmacological research. PMID- 14526383 TI - HIF-1 as a target for drug development. AB - Sensing and responding to fluxes in oxygen tension is perhaps the single most important variable in physiology, and animal tissues have developed a number of essential mechanisms to cope with the stress of low physiological oxygen levels, or hypoxia. Among these coping mechanisms is the response mediated by the hypoxia inducible transcription factor, or HIF-1. HIF-1 is an essential component in changing the transcriptional repertoire of tissues as oxygen levels drop, and could prove to be a very important target for drug development, as treatments evolve for diseases, such as cancer, heart disease and stroke, in which hypoxia is a central aspect. PMID- 14526385 TI - A guide to drug discovery. Protecting your inventions: the patent system. AB - Whether you work for a multi-national pharmaceutical company, a biotech start-up or a university, a knowledge of the patent system is essential for understanding how best to protect the fruits of your research. The aim of this article is to give an overview of what a patent is, how you might get one and the rights that a patent confers. PMID- 14526386 TI - Target discovery. AB - Target discovery, which involves the identification and early validation of disease-modifying targets, is an essential first step in the drug discovery pipeline. Indeed, the drive to determine protein function has been stimulated, both in industry and academia, by the completion of the human genome project. In this article, we critically examine the strategies and methodologies used for both the identification and validation of disease-relevant proteins. In particular, we will examine the likely impact of recent technological advances, including genomics, proteomics, small interfering RNA and mouse knockout models, and conclude by speculating on future trends. PMID- 14526387 TI - Quest for the best. AB - By combining innovative science with years of massive investment, drug makers seek to turn newly discovered chemicals into revolutionary blockbuster drugs that generate billions of dollars in revenue. So every year, they collectively spend tens of billions of dollars on the high-risk pursuit of the next Prozac or Viagra. But should they? This article analyses key themes around differentiation that we have found to be common among blockbusters, and examines the implications for creating future billion-dollar drugs. PMID- 14526388 TI - The Drosophila wing differentiation factor vestigial-scalloped is required for cell proliferation and cell survival at the dorso-ventral boundary of the wing imaginal disc. AB - Links between genes involved in development, proliferation and apoptosis have been difficult to establish. In the Drosophila wing disc, the vestigial (vg) and the scalloped (sd) gene products dimerize to form a functional transcription factor. Ectopic expression of vg in other imaginal discs induces outgrowth and wing tissue specification. We investigated the role of the VG-SD dimer in proliferation and showed that vg antagonizes the effect of dacapo, the cyclin-cdk inhibitor. Moreover, ectopic vg drives cell cycle progression and in HeLa cultured cells, the VG-SD dimer induces cell proliferation per se. In Drosophila, ectopic vg induces expression of dE2F1 and its targets dRNR2 and string. In addition vg, but not dE2F1, interacts with and induces expression of dihydrofolate reductase (DHFR). Moreover, a decrease in VG or addition of aminopterin, a specific DHFR inhibitor, shift the dorso-ventral boundary cells of the disc to a cell death sensitive state that is correlated with reaper induction and DIAP1 downregulation. This indicates that vg in interaction with dE2F1 and DHFR is a critical player for both cell proliferation and cell survival in the presumptive wing margin area. PMID- 14526389 TI - Life and death decisions by E2F-1. AB - Deregulation of the transcription factor E2F-1 is a common event in most human cancers. Paradoxically, E2F-1 has been shown to have the ability to induce both cell cycle progression and programmed cell death, leading potentially to both tumour-promoting as well as tumour-suppressive effects. Although the pathway to cell cycle progression seems straightforward with a number of growth-promoting E2F target genes having been described, the pathways to apoptosis are less well defined and more complex. The discovery that E2F-1 'knockout' mice are highly tumour prone has caused a recent surge in the number of reports relating to programmed cell death. This review focuses on these recent findings, highlighting the way in which they have increased our understanding of E2F-1-induced cell death, as well as indicating the questions that remain. Insight gained as to the role of this intriguing molecule in cancer and its potential for targeted therapy will also be discussed. PMID- 14526390 TI - Wedelolactone suppresses LPS-induced caspase-11 expression by directly inhibiting the IKK complex. AB - Caspase-11 is a key regulator of proinflammatory cytokine IL-1beta maturation and pathological apoptosis. Caspase-11 is not expressed in most tissues under normal condition, but highly inducible upon pathological stimulation such as in the presence of lipopolysaccharide (LPS). Here, we describe the identification and characterization of wedelolactone, a natural compound that inhibits LPS-induced caspase-11 expression in cultured cells by inhibiting NF-kappaB-mediated transcription. We demonstrate that wedelolactone is an inhibitor of IKK, a kinase critical for activation of NF-kappaB by mediating phosphorylation and degradation of IkappaBalpha. PMID- 14526391 TI - Missense mutations in hMLH1 and hMSH2 are associated with exonic splicing enhancers. AB - There is a critical need to understand why missense mutations are deleterious. The deleterious effects of missense mutations are commonly attributed to their impact on primary amino acid sequence and protein structure. However, several recent studies have shown that some missense mutations are deleterious because they disturb cis-acting splicing elements-so-called "exonic splicing enhancers" (ESEs). It is not clear whether the ESE-related deleterious effects of missense mutations are common. We have evaluated colocalization of pathogenic missense mutations (found in affected individuals) with high-score ESE motifs in the human mismatch-repair genes hMSH2 and hMLH1. We found that pathogenic missense mutations in the hMSH2 and hMLH1 genes are located in ESE sites significantly more frequently than expected. Pathogenic missense mutations also tended to decrease ESE scores, thus leading to a higher propensity for splicing defects. In contrast, nonpathogenic missense mutations (polymorphisms found in unaffected individuals) and nonsense mutations are distributed randomly in relation to ESE sites. Comparison of the observed and expected frequencies of missense mutations in ESE sites shows that pathogenic effects of >/=20% of mutations in hMSH2 result from disruption of ESE sites and disturbed splicing. Similarly, pathogenic effects of >/=16% of missense mutations in the hMLH1 gene are ESE related. The colocalization of pathogenic missense mutations with ESE sites strongly suggests that their pathogenic effects are splicing related. PMID- 14526393 TI - Intrahepatic cholestasis after liver transplantation. AB - Cholestasis is a common sequela of liver transplantation. Although the majority of cases remain subclinical, severe cholestasis may be associated with irreversible liver damage, requiring retransplantation. Therefore, it is essential that clinicians be able to identify and treat the syndromes associated with cholestasis. In this review, we consider causes of intrahepatic cholestasis. These may be categorized by time of occurrence, namely, within 6 months of liver transplantation (early) and thereafter (late), although there may be an overlap in their causes. The causes of intrahepatic cholestasis include ischemia/reperfusion injury, bacterial infection, acute cellular rejection, cytomegalovirus infection, small-for-size graft, drugs for hepatotoxicity, intrahepatic biliary strictures, chronic rejection, hepatic artery thrombosis, ABO blood group incompatibility, and recurrent disease. The mechanisms of cholestasis in each category and the clinical presentation, diagnosis, treatment, and outcome are discussed in detail. PMID- 14526392 TI - Microduplication 22q11.2, an emerging syndrome: clinical, cytogenetic, and molecular analysis of thirteen patients. AB - Chromosome 22, particularly band 22q11.2, is predisposed to rearrangements due to misalignments of low-copy repeats (LCRs). DiGeorge/velocardiofacial syndrome (DG/VCFS) is a common disorder resulting from microdeletion within the same band. Although both deletion and duplication are expected to occur in equal proportions as reciprocal events caused by LCR-mediated rearrangements, very few microduplications have been identified. We have identified 13 cases of microduplication 22q11.2, primarily by interphase fluorescence in situ hybridization (FISH). The size of the duplications, determined by FISH probes from bacterial artificial chromosomes and P(1) artificial chromosomes, range from 3-4 Mb to 6 Mb, and the exchange points seem to involve an LCR. Molecular analysis based on 15 short tandem repeats confirmed the size of the duplications and indicated that at least 1 of 15 loci has three alleles present. The patients' phenotypes ranged from mild to severe, sharing a tendency for velopharyngeal insufficiency with DG/VCFS but having other distinctive characteristics, as well. Although the present series of patients was ascertained because of some overlapping features with DG/VCF syndromes, the microduplication of 22q11.2 appears to be a new syndrome. PMID- 14526394 TI - Poor survival after liver retransplantation: is hepatitis C to blame? AB - Data from 1990 to 1996 suggest that the prevalence of hepatitis C virus (HCV) infection in repeated orthotopic liver transplantation (re-OLT) is increasing, and patient survival may be worse. Aims of the study are to: (1) assess the prevalence of HCV in re-OLT, (2) compare survival between primary OLT and re-OLT for HCV versus non-HCV diseases, and (3) evaluate Model for End-Stage Liver Disease (MELD) scores in re-OLT. The United Network for Organ Sharing database for adult patients undergoing primary OLT or re-OLT from January 1996 to June 2002 was analyzed. Patients with malignancy or those who underwent re-OLT within 30 days of primary OLT were excluded. A total of 22,120 primary OLTs and 2,129 re OLTs were performed. HCV was noted in 9,564 primary OLTs (43.2%) and 899 re-OLTs (42.2%). Overall 1, 3, and 5-year patient survival rates were 86%, 79%, and 73% for primary OLT, but 67%, 56%, and 52% for re-OLT (P <.001). Survival rates of patients with HCV at 1, 3, and 5 years were 86%, 76%, and 68% for primary OLT and 61%, 50%, and 45% for re-OLT (P <.001). Survival was less for patients with HCV compared with those with autoimmune hepatitis (AIH) and hepatitis B for re-OLT (P <.01). However, survival after re-OLT was no different for those with HCV than for those with all other causes. MELD scores between 11 and 20 were the most common for re-OLT. A marked decreased in survival was noted in all patients who underwent re-OLT with MELD scores greater than 25. HCV prevalence in OLT has reached a plateau in recent years. Survival after re-OLT is inferior to that for primary OLT, but re-OLT survival appears to have improved. Survival after re-OLT is lower in patients with HCV compared with those with AIH and hepatitis B, but no different than for those with most other liver diseases. Survival appeared worse in patients who underwent re-OLT with a MELD score greater than 25. PMID- 14526395 TI - Retransplantation for recurrent hepatitis C in the model for end-stage liver disease era: how should we or shouldn't we? PMID- 14526396 TI - Increased risk of cholestatic hepatitis C in recipients of grafts from living versus cadaveric liver donors. AB - Histologic injury caused by recurrent hepatitis C virus (HCV) has been reported in up to 90% of HCV-infected patients who undergo liver transplantation with a cadaveric graft. However, the natural history of HCV after living donor liver transplantation (LDLT) is not well described. We performed a retrospective analysis of 68 consecutive HCV-infected adult patients: 45 recipients of cadaveric grafts (CAD) were compared with 23 LDLT patients. Elevated serum transaminases, positive HCV RNA, and liver biopsy consistent with histologic evidence of HCV defined recurrence. When comparing CAD with LDLT, both the incidence of HCV recurrence and time to recurrence were not different. The overall incidence of severe sequelae of HCV recurrence, either cholestatic hepatitis, grade III-IV inflammation, and/or HCV-induced graft failure requiring retransplantation, was also not different when comparing CAD with LDLT. However, when comparing CAD versus LDLT, no CAD patient developed cholestatic hepatitis C, compared with 17% of LDLT who developed this complication (P =.001). Thus, in this patient population, the timing and incidence of HCV recurrence were not different when comparing CAD versus LDLT, but the incidence of cholestatic hepatitis was significantly greater in patients with HCV who underwent LDLT. PMID- 14526397 TI - Living donor liver transplantation in patients with chronic hepatitis C: timing is everything. PMID- 14526398 TI - Sequence variation in the hypervariable region 1 of hepatitis C virus and posttransplantation recurrent hepatitis. AB - Hepatitis C virus (HCV) shows remarkable genetic variation in both populations and individuals, in whom it circulates as quasispecies (QS). Sequence variation within an infected host has adaptive significance and reflects the modes and intensity of selection mechanisms operating on the virus. We investigated the sequence diversity of hypervariable region 1 of HCV in liver transplant recipients and correlated it with the recurrence of hepatitis. Twenty-six patients were considered during a 2-year period; all had graft reinfection, and 14 patients developed hepatitis recurrence. Cloned sequences were obtained from sera collected before or within 1 month after orthotopic liver transplantation (OLT) and at 3 and 24 months thereafter. Sequence diversity within single sera and over consecutive samples was analyzed quantitatively by matrix comparison and phylogenetic analysis. Propagation of viral QS in the graft was markedly dependent on individual factors. Viral QS in post-OLT sera were less complex and evolved slower compared with immunocompetent subjects with chronic hepatitis. Sequence variation was greater during the first 3 months post-OLT than during the remaining period. Genetic diversity within single samples was not related to hepatitis recurrence or other clinical features. Conversely, sequence diversity over consecutive samples was reduced in patients who experienced hepatitis recurrence, in particular, in those infected with genotype 1b and with an HLA-DR mismatched graft. Selection of viral sequences was markedly impaired in liver transplant recipients and tended to be greater early after OLT. Reduced sequence turnover correlated negatively with the outcome of graft reinfection. PMID- 14526399 TI - Hepatitis C virus quasispecies: misunderstood and mistreated? PMID- 14526400 TI - Outcomes of hepatitis C- and hepatitis B core antibody-positive grafts in orthotopic liver transplantation. AB - The use of hepatitis B core antibody (HBcAb)- and hepatitis C virus antibody (HCV+) liver grafts for transplantation in selected populations has not affected patient and graft survival. We reexamined the clinical outcomes of using these HBcAb+ and HCV+ grafts at our institution, in addition to studying recipients of combined HBcAb+/HCV+ grafts. We identified 377 patients who underwent transplantation for either hepatitis B and/or hepatitis C, or received both HBcAb+ and HCV+ grafts. Patient and graft survival at 5 years posttransplantation was 73% and 71%, respectively, in the HBcAb+ grafts compared with 81% and 75% in the HBcAb- grafts (P =.65; P =.94). For HCV+ grafts, patient and graft survival at 5 years posttransplantation was 89% and 73%, respectively, compared with 69% and 59% in the HCV- grafts; (P =.22; P =.77). The 5-year patient and graft survival rate in those who received combined HBcAb+/HCV+ grafts was 74% and 69%, respectively, and there was no statistical difference compared with the HBcAb+ and HCV+ grafts (P =.76; P =.90). The 5-year patient and graft survival rate in patients who received dual HBV prophylaxis with hepatitis B immunoglobulin (HBIg) and lamivudine was 88% and 84%, respectively, which was significantly higher than for patients who received single prophylaxis or no prophylaxis (P <.01; P =.02). Our study supports previous observations that patient and graft survival is not affected with the use of HBcAb+ and HCV+ grafts, and that dual prophylaxis with HBIg and lamivudine offers substantial survival benefits. Furthermore, the use of combined HBcAb+/HCV+ grafts did not impact patient or graft survival. This provides a potential new pool of donor livers that can be used for transplantation in select patients. PMID- 14526401 TI - Positive T lymphocytotoxic cross-match in living donor liver transplantation. AB - The influence of lymphocytotoxic cross-match on survival or acute rejection in living donor liver transplantation (LDLT) has not been well examined. We analyzed 133 consecutive adult LDLT cases and assessed patient survival and acute rejection rates. Patients with a positive T lymphocytotoxic cross-match (n = 12) had a significantly higher chance of rejection within 6 weeks of LDLT (67% versus 28%, P <.001). All of the rejection episodes were successfully treated with bolus methylprednisolone therapy or anti-T cell monoclonal antibody. T lymphocytotoxic cross-match-positive grafts had no influence on patient survival (79% versus 90% at 3 years, P =.91). The results show that a positive cross-match graft should not be considered a contraindication for LDLT. PMID- 14526402 TI - Results of combined and sequential liver-kidney transplantation. AB - Experience with combined liver-kidney transplantation (L-KTx) has increased, but controversy regarding this procedure continues because the indications are not clearly defined yet. Between 1984 and 2000, 38 patients underwent simultaneous L KTx and 9 patients underwent sequential transplantation, receiving either a liver before a kidney or a kidney before a liver. Main indications for a simultaneous procedure were polycystic liver-kidney disease with cirrhosis and coincidental renal failure. The main indications for sequential procedure were cirrhosis caused by viral infection for the liver and glomerulonephritis for the kidneys. Outcomes in these patients were evaluated retrospectively. Regarding simultaneous transplantation, 28 (73.7%) long-term survivors were followed up for 0.7 to 12.5 years. Currently, 24 (63.2%) patients are alive with good liver function. Fourteen patients died; 10 patients died in the early postoperative phase because of septic complications, and most of them were cirrhotic with a poor preoperative clinical status. Currently, 2 of the surviving patients (8%) have returned to dialysis, 4 (17%) have reduced renal function, and 18 (75%) have good renal function. Five liver and 2 kidney retransplantations were performed during the follow-up. In cases of sequential grafting, patients undergoing kidney transplantation in the presence of a previously transplanted stable liver did better than those who underwent liver transplantation after kidney transplantation. When liver transplantation was performed early and electively before substantial worsening, combined L-KTx is a safe procedure offering excellent long-term palliation. PMID- 14526403 TI - Renal function improves in liver transplant recipients when switched from a calcineurin inhibitor to sirolimus. AB - Sirolimus (Rapamune; Wyeth-Ayerst, Philadelphia, PA) is a newer immunosuppressive drug with no known acute or chronic nephrotoxic effects; however, limited data are available in liver transplant recipients. We prospectively evaluated changes in renal function in liver transplant recipients after conversion from a calcineurin inhibitor to sirolimus monotherapy. We measured serial serum creatinine levels in liver transplant recipients with chronic nephrotoxicity caused by calcineurin inhibitors before and after conversion to sirolimus therapy. Estimated glomerular filtration rate (eGFR) was calculated from the Modification of Diet in Renal Disease formula. Change in eGFR over time, incidence of acute hepatocellular rejection, and adverse events while being administered sirolimus monotherapy were recorded. Mean interval between liver transplantation and initiation of sirolimus therapy was 310 weeks (range, 9 to 780 weeks). Of 21 patients included in our study, 18 patients were converted to sirolimus monotherapy and 3 patients were switched to sirolimus and low-dose steroid therapy. Patients were followed up for a mean of 66.8 +/- 38.9 (SD) weeks after conversion. Renal function improved in 71% of patients (15 of 21 patients). Median eGFR improved significantly from 34 mL/min/1.73 m2 at the time of conversion to 43 mL/min/1.73 m2 at the last follow-up (27% increase in eGFR; P = 001). Median monthly change in eGFR was from -0.25 mL/min/1.73 m2 pre-sirolimus therapy to +1.28 mL/min/1.73 m2 post-sirolimus therapy (P =.09). Adverse events were mostly mild and self-limited. Only 1 patient developed biopsy-proven acute cellular rejection, which was treated with sirolimus and mycophenolate mofetil. Two patients discontinued sirolimus therapy because of toxicity (oral ulceration, 1 patient; interstitial pneumonitis, 1 patient). Renal function improved significantly in the majority of liver transplant recipients with renal insufficiency caused by calcineurin inhibitors when converted to sirolimus therapy. Sirolimus monotherapy provided adequate immunosuppression with a low incidence of acute cellular rejection and minimal adverse events. PMID- 14526404 TI - Recurrence of primary biliary cirrhosis after liver transplantation: Histologic estimate of incidence and natural history. AB - The goals of this study were to determine the rate of recurrent primary biliary cirrhosis (PBC) after orthotopic liver transplantation (OLT) based on strict morphologic criteria and to evaluate histologic progression of recurrent PBC over time. Strict criteria for PBC recurrence were established as the presence of a florid duct lesion or destructive lymphocytic cholangitis within a dense portal infiltrate. Of the 784 OLTs performed at the Mayo Clinic during the first 12 years of the program, 100 met criteria for the PBC study group, and 35 met criteria for the control group. Strict histologic criteria for recurrent PBC were observed in 17 of 100 (17%) study patients (14 with florid duct lesion, 3 with destructive lymphocytic cholangitis within dense portal infiltrate). Mean follow up for the PBC group was 4.7 years (range, 1.0 to 13.8). Mean time to recurrence was 3.7 years (median, 3.1; range, 0.3 to 7.9). In those who met strict criteria for recurrent PBC, 2 of 17 progressed to septal fibrosis (stage 3). No florid duct lesions, destructive lymphocytic cholangitis, or septal fibrosis were observed in the control group. Other less specific morphologic features of PBC (portal infiltrates, plasma cells, dense lymphoplasmacytic infiltrates, and lymphocytic cholangitis) were also evaluated in the course of this study. Based on strict criteria, a conservative histologic estimate of the rate of recurrent PBC is 17% with a mean of 4.7 years of follow-up. When criteria for histologic recurrence are expanded to include moderate lymphocytic cholangitis with lymphoplasmacytic portal infiltrate, the recurrence rate of PBC is estimated as 26%. PMID- 14526405 TI - Generation of hepatocytes from cultured mouse embryonic stem cells. AB - Embryonic stem (ES) cells are pluripotent cells derived from the inner cell mass of fertilized blastocysts in vitro. ES cells can be induced to undergo differentiation into potentially all cell types. The aim of this study is to examine the differentiating potential of mouse ES cells into hepatocytes in the presence of retinoic acid (RA), hepatocyte growth factor (HGF), and beta-nerve growth factor (beta-NGF). RA, HGF, and beta-NGF were added to the cell culture. Hepatocyte induction was confirmed morphologically, as well as biochemically, through immunohistochemical assays of alpha1-antitrypsin (alpha1-AT) and alfafetaprotein (AFP) expression and reverse-transcriptase polymerase chain reaction tests for the presence of albumin, transthyretin, glucose 6 phosphates, hepatic nuclear factor 4, and SAPK/ERK kinase-1 (SEK1) messenger RNA, produced only by functioning hepatocytes. Fifteen days after the addition of HGF and beta NGF to the cell culture, many epithelioid cells were noticed. alpha1-AT, AFP, albumin, transthyretin, glucose 6 phosphates, hepatic nuclear factor 4, and SEK1 messenger RNA expression also was detected, indicating successful ES cell differentiation into functioning hepatocytes. However, in the presence of RA alone, only transthyretin messenger RNA was positive, whereas no other expression pertaining to functioning hepatocytes could be detected. In the presence of HGF and beta-NGF, mouse ES cells can differentiate into functioning hepatocytes, whereas RA function is limited. PMID- 14526406 TI - Making hepatocytes from stem cells: where are we? PMID- 14526407 TI - Death from donor-transmitted malignancy despite emergency liver retransplantation. AB - Transplantation of organs procured from donors with malignancies identified subsequent to implantation presents a significant dilemma regarding the optimal management strategy to simultaneously minimize the risk for cancer transmission and recipient morbidity. In this report, we present a patient who underwent orthotopic liver transplantation for hepatitis B cirrhosis. The donor had no previous history of cancer. On autopsy, enlarged mediastinal lymph nodes led to the discovery of a 1-cm lung tumor. Histological examination showed pulmonary adenocarcinoma with metastatic mediastinal disease. Despite urgent retransplantation within 7 days, the recipient developed metastatic pulmonary adenocarcinoma diagnosed 11 months after transplantation and died soon thereafter. Analysis of short tandem repeat regions of chromosomal DNA from the recipient, the 2 liver donors, and the posttransplantation tumor corroborates that the first donor was the source of the malignancy. This case of donor transmitted malignancy underscores the need for vigilance by the procuring surgeon in identifying potential malignancy during organ retrieval and use of a full autopsy on selected donors after organ procurement. PMID- 14526409 TI - Pharmacogenomics in liver transplantation. PMID- 14526410 TI - Arterial anatomy unsuitable for a right liver donation. PMID- 14526408 TI - Enhanced expression of enterocyte P-glycoprotein depresses cyclosporine bioavailability in a recipient of living donor liver transplantation. AB - We evaluated levels of intestinal expression of absorptive-barrier P-glycoprotein (PGP) and cytochrome P-450 IIIA4 (CYP3A4) and immunosuppressant therapy in a patient who underwent living donor liver transplantation (LDLT) and received a second living donor liver transplant after chronic rejection of the first. PGP and CYP3A4 expression were measured using part of a Roux-en-Y limb. After the first LDLT, the concentration-dose ratio of orally administered tacrolimus was 159.8 +/- 125.3 (average +/- SD of 32 different days), similar to the average for 46 recipients of living donor liver transplants in our hospital (161.3 +/- 88.1). However, the recipient required very large oral doses of cyclosporine (703.9 +/- 385.4 mg/d, average +/- SD of 13 different days) after the second LDLT. Although intestinal PGP level was increased markedly at the second LDLT, CYP3A4 level was decreased. In addition, levels of messenger RNA expression of several gene products related to the local inflammation, such as cyclooxygenase 2, interleukin 1beta (IL-1beta), IL-2, IL-6, IL-8, IL-10, and tumor necrosis factor-alpha, were increased. These results suggest that hepatic failure after LDLT, including chronic rejection and/or cholangitis, was accompanied by upregulation of intestinal PGP expression, which could depress the bioavailability of the immunosuppressant. PMID- 14526412 TI - Executive summary from the Intraoperative Advisory Council on Donation After Cardiac Death of the United Network for Organ Sharing: practice guidelines. AB - The number of organs recovered from asystolic donors is less than anticipated and is explained partially by negative attitudes held by health care providers. To understand the reasons for these beliefs and find solutions, the United Network for Organ Sharing under contract with the Department of Health and Human Services convened the Intraoperative Advisory Council on Donation After Cardiac Death in September, 2001. The Council found that, unlike other medical specialties, operating room health care providers were uncertain of their roles and duties in the care of donors declared dead with cardiopulmonary criteria, known as donation after cardiac death. They were reluctant to care for terminally ill patients in whom death was an expected outcome. Council members deliberated these issues, seeking solutions to repatriate operating room health care providers with the national effort to provide reliable and compassionate care to organ donors and their families. The Council requested the construction of practice guidelines, believing that the structure provided by guidelines will improve health provider confidence in donation after cardiac death and thus improve the quality of care. Physician and nonphysician health care providers from the operating room met to create the Guidelines for the United Network for Organ Sharing, which they believe will improve the quality of care of asystolic organ donors. PMID- 14526413 TI - History of the International Liver Transplantation Society. PMID- 14526415 TI - Relationship between apoptosis and E-cadherin expression in bronchial epithelium of smoking mouse. AB - To investigate whether apoptosis is associated with cell adhesion in bronchial epithelium, and whether it contributes to the kinetics of injury and repair of surface epithelia, this study was performed for E-cadherin expression by using immunohistochemistry technique and for apoptosis by TUNEL method. An animal model of smoking was used for this study. The results showed that epithelial cells with membrane anchored E-cadherin decreased remarkably at several time points during 6 months of exposure to smoke (P < 0.01) and then restored to normal level. This fluctuation was associated exclusively with the alteration in number of apoptotic cells (P < 0.01). There was no significant difference in activation of nuclear transcription factor NF-kappa B among groups (P > 0.05). All these suggested that apoptosis is associated with E-cadherin expression in bronchial epithelium of smoking mouse. PMID- 14526414 TI - Comparative study on the immunogenicity between recombinant MS-Sj26GST vaccine and recombinant BCG-Sj26GST vaccine in Schistosoma japonicum. AB - The BALB/c mice were immunized with rMS-Sj26GST and rBCG-Sj26GST vaccine in Schistosoma japonicum by subcutaneous injection. After they were immunized for 8 weeks, the eyeballs were removed to get blood and macrophages of abdominal cavity and spleen cells were harvested. The lymphocytic stimulating index (SI) was used to measure the cellular proliferating ability and NO release was used to measure the phagocytic activity of the macrophages. By using ELISA kit, the levels of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) in serum and the splenic lymphocytic cultured supernatant were detected. The results showed that after the mice were immunized with 10(6) CFU of rMS-Sj26GST and rBCG-Sj26GST vaccine separately by subcutaneous injection, proliferating ability of splenic lymphocytes in the mice showed no difference (P > 0.05), but both were significantly increased as compared with that in the control group (P < 0.05); The contents of NO in the intraperitoneal macrophages of rMS-Sj26GST vaccine group were significantly lower than in the control group (P < 0.001) and rBCG Sj26GST vaccine group (P < 0.01); The levels of serum IL-2 in the rMS-Sj26GST vaccine group were significantly increased as compared with that in the control group (P < 0.001), vector group (P < 0.01) and rBCG-Sj26GST vaccine group (P < 0.05); The contents of serum IFN-gamma in the rMS-Sj26GST vaccine group were significantly increased as compared with that in the control group (P < 0.01) and rBCG-Sj26GST vaccine group (P < 0.05), The contents of IFN-gamma in the cultured supernatant were significantly lower than those of rBCG-Sj26GST vaccine group (P < 0.001), but were significantly increased as compared with that in the control group (P < 0.01). It was indicated that both vaccines could enhance the immune response of the mice, but rMS-Sj26GST vaccine had stronger immunogenicity than rBCG-Sj26GST vaccine. PMID- 14526416 TI - Expression of macrophage inflammatory protein 1 alpha in the endothelial cells exposed to diamide. AB - In order to study whether the endothelial cells (ECs) with lipid peroxidation induced by diamide can express and secrete macrophage inflammatory protein 1 alpha (MIP-1 alpha), the expression of MIP-1 alpha protein in the cells was detected by cell enzyme-linked immunosorbent assay (ELISA) and that of MIP-1 alpha mRNA was determined by cell in situ hybridization and nuclease S1 protection assay after the ECs were exposed to different concentrations of diamide for 4 h. The chemotactic activity of MIP-1 alpha was tested by micropore filter method using modified Boyden chambers. Cell ELISA showed that the expression of MIP-1 alpha protein in endothelial cells exposed to 1 mumol/L, 5 mumol/L and 10 mumol/L diamide was 1.9-fold, 2.3-fold and 1.7-fold respectively as much as that in the control cells, which was statistically significant by analysis of variance. In situ hybridization revealed that the mRNA expression of ECs treated with 1 mumol/L, 5 mumol/L and 10 mumol/L diamide was 1.3-fold, 3.0 fold and 1.7-fold as much as that in the control group, which had statistical significance (F = 188.93, P < 0.01). The mRNA expression in 5 mumol/L dimide treated ECs, measured by nuclease S1 protection assay, was 3.4-fold as much as that in the control group (t = 8.70, P < 0.05). Chemotactic response(99.50 +/- 4.31 microns) to the culture medium conditioned by 5 mumol/L diamide treated ECs, which was stronger than that(66.47 +/- 3.25 microns) conditioned by the ECs (F = 404.31, P < 0.05), was significantly decreased (F = 192.25, P < 0.05) after adding MIP-1 alpha antibody. It suggests that diamide, a lipid peroxidation inducer, could stimulate ECs to produce high level of MIP-1 alpha, and might play an important role in atherogenesis by promoting the migration of peripheral blood monocytes into arterial intima. PMID- 14526417 TI - Inhibitory effect of recombinant endostatin on angiogenesis and tumor growth of hepatoma. AB - To study the influence of recombinant endostatin on angiogenesis and tumor growth of mice H22 hepatoma, tumor models were constructed by injecting H22 hepatoma cells into the leg muscle of mice. Recombinant endostatin was produced by gene engineering in E. coli. The recombinant protein was injected subcutaneously to treat transplanted hepatoma faraway. The weight of tumors was measured, and the changes of necrosis of tumor cells and vessel density were observed by immunohistochemistry. The results suggested that the growth of hepatoma models transplanted in the muscle of legs was suppressed by recombinant endostatin. The density of vascularity was decreased, but the necrosis of tumor cells increased. The inhibitory effect of recombinant endostatin on angiogenesis and tumor growth of hepatoma was not affected after chemotherapy. PMID- 14526418 TI - A nonradioactive method for detecting DNA-binding activity of nuclear transcription factors. AB - To determine the feasibility of a nonradioactive electrophoresis mobility shift assay for detecting nuclear transcription factor, double-stranded oligonucleotides encoding the consensus target sequence of NF-kappa B were labelled with DIG by terminal transferase. After nuclear protein stimulated with phorbol 12-myristate 13-acetate (PMA) or PMA and pyrrolidine dithiocarbamate (PDTC) electrophoresed on 8% nondenaturing poliacrylamide gel together with oligeonucleotide probe, they were electro-blotted nylon membrane positively charged. Anti-DIG-AP antibody catalyzed chemiluminescent substrate CSPD to image on X-film. The results showed that nuclear proteins binded specifically to the NF kappa B consensus sequence in the EMSA by chemiluminescent technique method and the activity of NF-kappa B in PMA group was more than that in PMA + PDTC group. It is suggested that detection of NF-kappa B by EMSA with chemiluminescent technique is feasible and simple, which can be performed in ordinary laboratories. PMID- 14526420 TI - Synthesis of CCK-8 tetrapeptide fragment by enzymatic method. AB - The enzymatic synthesis of a tetrapeptide Phac-Met-Gly-Trp-Met-OEt, a fragment of the cholecystokinin C-terminal octapeptide CCK-8, was reported. This fragment was synthesized by coupling Phac-Met-OEt with Gly-OMe, Trp-OMe and Met-OEt successively. These three steps were catalyzed by alpha-chymotrpsin, Papain and alpha-chymotrpsin respectively. The results of FAB-MS showed that all the products had the correct molecular mass. PMID- 14526419 TI - Effects of lipoteichoic acid induced delayed preconditioning on ischemia reperfusion injury in isolated rat hearts. AB - To explore the potential of lipoteichoic acid (LTA) induced cardioprotection against ischemia-reperfusion (I/R) injury in isolated rat hearts and whether endogenous nitric oxide (NO) participates in the protection, the rats were pretreated with LTA (1 mg/kg, i.p.) 24 h before the experiment, and the isolated hearts were subjected to 30 min no-flow normothermic global ischemia and 60 min reperfusion after a 20-min stabilization period by the langendorff method. Cardiac functions were evaluated at the end of stabilization, and at 30 min, 60 min of reperfusion. The amounts of MB isoenzyme of creatine kinase (CK-MB), lactate dehydrogenase(LDH) and total NO oxidation products in the coronary effluent were measured spectrophotometrically at the end of reperfusion. It was revealed that pretreatment with LTA could significantly improve the recovery of cardiac function, reduce the release of CK-MB and LDH, and increase the concentrations of NO in coronary effluent. The protective effects were abrogated by pretreatment of the rats with L-NAME. It was concluded that LTA could induce the delayed cardioprotection against I/R injury, and endogenous NO may be involved in the mechanisms. PMID- 14526421 TI - Immunophenotype and ultrastructure of B-cell lymphoproliferative disorder with cytoplasmic projection. AB - To identify the knowledge of rare lymphoproliferative disorder, the clinical and biological features of three kinds of lymphoproliferative disorders with cytoplasmic projections were compared. The clinical manifestations, ultrastructure and immunophenotype were analyzed. The results showed that hairy cell leukemia (HCL), splenic lymphoma with villous lymphocyte (SLVL) and hairy cell leukemia-variant (HCL-V) had some common characters including splenomegaly, peripheral blood and bone marrow infiltration by villous lymphocyte and B lymphocyte immunophenotype; but these three disorders had specific features respectively. It was concluded that overall analysis of clinical and laboratory features might be contributive to the differential diagnosis of these three disorders. PMID- 14526422 TI - Distribution of variant genotypes of Fc gamma receptor IIIa in healthy Chinese population of Zhengzhou City. AB - To investigate the distribution of variant genotypes of Fc gamma receptor IIIa (Fc gamma R IIIa) in healthy Chinese population of Zhengzhou city, genomic DNA was extracted from peripheral blood of healthy donators. The genotypes of Fc gamma R IIIa-158 were determined by nested polymerase chain reaction (PCR) in 137 healthy people in Zhengzhou city. The results showed that frequencies of variant genotypes FF, VV and VF were 42.3%, 48.9% and 8.8% respectively. The distribution of Fc gamma R IIIa-158 in healthy Chinese population of Zhengzhou city was polymorphic and different from that of African Americans (AA) and Caucasian Americans (CA). PMID- 14526423 TI - Mutation identification in a 5-generation pedigree with autosomal dominant retinitis pigmentosa. AB - An extended 5-generation family has been investigated in which 32 of the 111 family members were diagnosed as having retinitis pigmentosa (RP). The proband was a 58-year old male in whom night-blindness was first observed in early childhood, with almost loss of vision by 52 years of age. The symptoms observed in other family members included night-blindness, impaired vision and visual field loss. Dementia, digital abnormalities, deaf-mutism and mental retardation were variously diagnosed in a number of individuals with RP. The affected and unaffected family members were tested for mutations in a range of candidate genes. The 8 exons of three candidate genes have been analyzed by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and DNA sequencing techniques. A novel mutation was identified in the rhodopsin gene at codon 52 of exon 1 (TTC-TAC) that resulted in a substitution of Phe to Tyr. PMID- 14526424 TI - Correlation between cyclin A gene expression in adult patients with acute leukemia and drug resistance. AB - In order to investigate the relationship between the expression of cyclin A and drug resistance in adult patients with acute leukemia (AL), the mRNA expression of cyclin A, mdr1, Top II alpha, bcl-2 was detected in 64 adult patients with AL and 20 normal controls by semi-reverse transcription polymerase chain reaction (semi-RT-PCR). It was found that the cyclin A and Top II alpha mRNA expression levels in drug resistant group were significantly lower than in sensitive group (P < 0.01). Under the same experimental condition no cyclin A mRNA expression was detectable in all normal controls. The mdr1 and bcl-2 mRNA expression levels in resistant group were significantly higher than in sensitive group (P < 0.01), cyclin A and Top II alpha gene expression levels were closely correlated (rs = +0.794, P = 0.000, n = 64) in all AL patients, but cyclin A was not correlated with mdr1 and bcl-2 gene expression levels. In drug resistant group there was a negative correlation between the gene expression levels of cyclin A and mdr1 (rs = -0.337, P = 0.029). The 10 AL patients with positive lower expression of both cyclin A and Top II alpha were all resistant to drugs. Logistic regression of Binary analysis showed the correlation between the lower expression of cyclin A and drug resistance. It was concluded that lower expression of cyclin A gene might be an unfavorable prognostic factor for patients with AL, and detection of both cyclin A and Top II alpha gene expression would predict drug resistance in AL patients. PMID- 14526425 TI - Effect and comparison of sodium butyrate and trichostatin A on the proliferation/differentiation of K562. AB - In order to explore the molecular mechanisms of sodium butyrate and trichostatin A on K562 cell proliferation/differentiation, K562 cells were grown in the absence or presence of sodium butyrate or trichostatin A. The percentage of viable cells was determined by trypan blue exclusion. Differentiation was determined by nitro-blue tetrazolium (NBT) reduction and cell surface adhesion molecules analyzed by FACS. Cell cycle distribution was studied after DNA staining by propidium iodide. Cell cycle regulatory proteins were detected by Western blot and reverse transcription-polymerase chain reaction. The results showed that sodium butyrate blocked cells mainly at the G0/G1 phase of the cell cycle, whereas trichostatin A arrested the cells at G2 phase. Sodium butyrate could down-regulate the mRNA expression of cyclin D1, but not affect its protein expression; down-regulate the protein expression of cyclin D3, but not affect its mRNA expression. Trichostatin A showed similar effects on cyclin D1 and D3 as sodium butyrate. Both sodium butyrate and trichostatin A could stimulate p21 expression of K562 cells at mRNA and protein levels. It may be concluded that sodium butyrate and trichostatin A could promote the proliferation/differentiation of the K562 cells, which might be contributed to the induced expression of cyclin D3 and p21 proteins. PMID- 14526426 TI - Induction of increased intracellular calcium in astrocytes by glutamate through activating NMDA and AMPA receptors. AB - To study the effect of glutamate on the intracellular calcium signal of pure cultured rat astrocytes and the role of NMDA and AMPA receptors in the procedure, the change of calcium signal was investigated by monitoring the fluctuation of intracellular Ca2+ concentration ([Ca2+]i) on the basis of Fura-2 single cell fluorescent ratio (F345/F380). The changes in the effect of glutamate on the intracellular calcium signal were observed after blockage of NMDA and (or) AMPA receptors. It was found that L-glutamate could induce an increased [Ca2+]i in most of the cells in concentration- and time-dependent manner. D-(-)-2-amino-5 phosphonopentanoic acid (D-AP-5, a selective antagonist of the NMDA receptor) and 6-cyano-7-nitroquinoxaline-2, 3-dione (CNQX, a selective antagonist of the AMPA receptor) could abolish the effects of NMDA and AMPA respectively. The treatment of D-AP-5 and CNQX simultaneously or respectively could attenuate the effect of L glutamate at varying degrees. All these indicated that glutamate could modulate intracellular Ca2+ of pure cultured rat astrocytes through different pathways. The activation of NMDA and AMPA receptors took part in the complex mechanisms. PMID- 14526427 TI - Effect of levodopa chronic administration on behavioral changes and fos expression in basal ganglia in rat model of PD. AB - To study behavioral character and changes of neuronal activity in the basal ganglia of rat model of levodopa-induced dyskinesia, unilateral 6-hydroxydopamine lesioned rat model of Parkinson disease (PD) was treated with levodopa/benserazide twice daily for 4 weeks and the behavior observed on the 1st, 3rd, 4th, 7th, 9th, 10th, 14th, 21st and 28th day. The animals were sacrificed and immunohistochemical technique was used to measure the changes of Fos expression in the caudate putamen (CPU), globus pallidus (GP) and sensorimotor area of cerebral cortex 2 h after the last treatment. The results showed that pulsatile treatment with a subthreshold dose of levodopa gradually induced abnormal involuntary movement (AIM), including stereotypy (limb dyskinesia, axial dystonia and masticatory dyskinesia) towards the side contralateral to the dopamine-denervated striatum and increased contraversive rotation. The motor pattern of each subtype was highly stereotypic across individual rats, and the proportion of each subtype was not consistent among individual rats. Fos positive nuclei in the CPU and GP were increased by levodopa acute administration, and more remarkably in the CPU, but not in the cerebral cortex. After repeated levodopa treatment. Fos positive nuclei were reduced remarkably in the CPU, but were increased in the GP and cerebral cortex. It was concluded that the neural mechanisms underlying levodopa induced AIM in rat model of PD was very similar to those seen in levodopa-induced dyskinesia (LID) in PD patients and MPTP-lesioned monkeys, and increased striatopallidal neuronal activity might be involved in occurrence of LID. PMID- 14526428 TI - Relationship between levels of serum C-reactive protein, leucocyte count and carotid plaque in patients with ischemic stroke. AB - In order to study the relationship between serum C-reactive protein (CRP) levels, leukocyte count and carotid plaque in patients with ischemic stroke, carotid duplex examination was performed by high-definition imaging (HDI) 5000 triplex system. Serum CRP was measured by nephelometry within 72 h after index ischemic stroke. A lesion was considered a plaque in the presence of a maximum intimal medial wall thickness (IMT) 1.2 mm. Results of carotid ultrasonography were divided into two groups: M1, normal (IMT < 1.2 mm) and M2, abnormal (IMT > or = 1.2 mm). The results showed that the mean age of M2 was significantly older than that of M1 (69.7 +/- 10.4 versus 62.5 +/- 9.6, P = 0.001). The patients with hypertension and diabetes mellitus (78%, 35% respectively) in M2 were significantly more than those (52%, 18% respectively) in M1 (P < 0.01, P < 0.05). There were 32 (65%) patients with elevated CRP levels in M2, but 33 (46%) patients with elevated CRP levels in M1, with the difference being significant between the two groups (P < 0.05). The levels of serum glucose and leukocyte count (8.1 +/- 5.5, 10.3 +/- 4.0, respectively) in abnormal CRP group were significantly higher than that of normal CRP group (6.4 +/- 2.8, 8.7 +/- 3.4) (P < 0.05, P < 0.05); elevated CRP levels was found in 42 (62%) patients with territory infarction and 23 (43%) patients with lacunar infarction respectively, with the difference being significant between these two groups (P < 0.05). It was concluded that the elevation of CRP levels was an significant clinical index for carotid plaque in patients with acute cerebral infarction. PMID- 14526429 TI - Effect of technetium-99 conjugated with methylene diphosphonate on IgM-RF, IgG-RF and IgA-RF. AB - To explore the effect of technetium-99 conjugated with methylene diphosphonate (99 TcMDP) on IgM-RF, IgG-RF and IgA-RF (RFs), 47 cases were selected for study, including 33 patients with rheumatoid arthritis (RA) and 15 patients with joint pain/arthritis. After 99Tc-MDP for drips model being given to the patients by intravenous drip 0.2 g daily for 5 days, the injection A and B models of 99Tc-MDP were used to the patients by intravenous injection one set daily for 10 days, that was one course of treatment. The next course started after 10 days. Each case used it from 2 to 4 courses of treatment. The RFs in serum were determined by the method of enzyme-linked immunoabsorption assay (ELISA) before and after 2 and 4 courses of treatment. In the patients with RA, the concentrations of IgM-RF were 296.2 +/- 108.4 IU/ml, 189.5 +/- 92.3 IU/ml and 107.8 +/- 72.5 IU/ml; the concentrations of IgG-RF were 325.6 +/- 126.2 IU/ml, 209.7 +/- 98.2 IU/ml and 160.2 +/- 80.8 IU/ml; the concentrations of IgA-RF were 330.4 +/- 136.3 IU/ml, 210.7 +/- 89.2 IU/ml and 148.8 +/- 72.2 IU/ml before and after 2 and 4 courses of treatment, respectively. The concentrations of the above RFs were significantly lower after 2 and 4 courses than those before treatment (P < 0.05 and P < 0.01). There was no significant difference in RFs concentrations in the patients with joint pain/arthritis before and after use of 99Tc-MDP. In the patients with positive RFs before treatment, the RFs concentrations were decreased significantly after 2 and 4 courses of treatment (P < 0.05 and P < 0.01). There was no obvious change of RFs concentrations in the patients with negative RFs after treatment of 99Tc-MDP. It was concluded that 99 Tc-MDP could obviously reduce the abnormally high concentrations of RFs, but not influence the normal RFs, which indicated that 99Tc-MDP has an important effect on controlling the activities of RA. PMID- 14526430 TI - Effects of mycophenolate mofetil on renal interstitial fibrosis after Unilateral ureteral obstruction in rats. AB - To investigate the effects of mycophenolate mofetil (MMF) on the process of renal interstitial fibrosis, unilateral ureteral obstruction (UUO) model was established in rats. Twenty Sprague-Dawley rats underwent UUO and received vehicle (n = 10) or MMF (20 mg.kg-1.d-1, by daily gastric gavage, n = 10) during a period of 5 days following surgery, and the additional 10 rats were served as sham-operated group. The rats were killed 5 days after surgery. Immunohistochemistry was performed on renal tissue for proliferating cell nuclear antigen (PCNA), alpha-smooth muscle actin (alpha-SMA) and type I and III collagen (col I, col III). Histological studies were also done by MASSON staining. Five days after surgery, proliferating cells in tubules, interstitium as well as interstitial myofibroblast (MyoF) infiltration and interstitial col I, col III deposition were all significantly reduced by MMF treatment. MMF also alleviated the histological changes of UUO rats. These results suggested that the reduction of interstitial MyoF infiltration may be an important event by which MMF prevents renal injury caused by UUO and MMF could be used to limit the progression of renal fibrosis. PMID- 14526431 TI - Expression of HO-1 in chronic renal insufficiency. Rat kidney and implication. AB - The expression, activity and clinical implication of heme oxygenase-1 (HO-1) in the chronic renal insufficiency (CRI) rat kidney and its mechanism were investigated. The 5/6 nephrectomized rats were assigned to sham operation group, CRI group and Hemin group. At the 8th week after second operation, blood pressure, urinary protein, serum creatinine (Scr) and BUN were measured. Renal pathologic changes were observed. The activity of HO and contents of erythropoietin (EPO) in serum and renal tissue were determined. Immunohistochemistry was used to detect the expression and distribution of HO-1 in the CRI rat kidney. As compared with CRI group, the urinary protein, blood pressure, Scr and BUN in Hemin group were reduced significantly (P < 0.05). The glomerular mesangial proliferation, inflammatory cellular infiltration of renal interstitium and interstitial fibrosis were ameliorated significantly. Immunohistochemistry and measurement of HO-1 activity revealed that the expression and activity of HO-1 was decreased in renal tissues and increased in serum in CRI group as compared with normal rats. HO-1 distributed mainly in tubular epithelial cells. The EPO contents in Hemin group were significantly higher than in CRI group. Through up-regulating the EPO level in serum and renal tissues, HO-1 retards the progression of CRI. PMID- 14526432 TI - Role of adrenomedullin in congenital heart disease associated with pulmonary hypertension. AB - The changes of adrenomedullin (ADM), endothelin-1 (ET-1) and nitric oxide (NO) levels before and after operation in congenital heart disease (CHD) associated with pulmonary hypertension (PH) were observed in order to investigate their role in CHD with PH and their clinical significance. The CHD patients were divided into 3 groups according to pulmonary artery systolic pressure (PASP): Non-PH group: PASP < or = 30 mmHg (n = 11); mild-PH group: PASP 31-49 mmHg (n = 10); moderate or severe-PH group: PASP > or = 50 mmHg (n = 12). The control group consisted of 15 health children. Plasma ADM, ET-1 and NO levels were determined by radioimmunoassay and colorimetry methods. The correlation between ADM and ET 1, NO, PASP was analyzed. The changes in plasma ADM, ET-1 and plasma NO on the 7th day after operation among the groups were compared. The results showed that plasma ADM levels in non-PH group were significantly higher than that in control group (P < 0.05), but there was no significant difference in ET-1 and NO levels between the two groups (P > 0.05). ADM and ET-1 levels in mild-PH group were significantly elevated as compared with those in non-PH group (both P < 0.05), but NO levels were decreased (P < 0.05). ADM and ET-1 levels in moderate or severe-PH groups were increased as compared with those in mild-PH group (both P < 0.01), but NO level significantly declined (P < 0.05). On the 7th day after operation, plasma ADM and ET-1 levels in PH group were significantly decreased (P < 0.05, P < 0.01) as compared with those before operation, but there was no significant difference in NO levels (P > 0.05). But NO levels in non-PH group were significantly increased (P < 0.05). Plasma ADM levels in CHD were positively correlated with PASP and ET-1 (r = 0.77, P < 0.01; r = 0.82, P < 0.01), negatively correlated with NO (r = -0.56, P < 0.05). It was concluded that during the progression of PH in the cases of CHD, plasma ADM, ET-1 and NO might play an important role in the development of PH. The increased ADM may represent a compensatory mechanism. It can interact with NO and ET-1 to regulate pulmonary circulation in the pathophysiology of PH with CHD. ADM may be involved in the defence mechanism against further increase of pulmonary arterial pressure. ADM could be used as a reliable indicator of the severity of CHD associated PH. PMID- 14526433 TI - Results of invasive electrophysiologic evaluation in 268 patients with unexplained syncope. AB - In order to assess the diagnostic value of invasive electrophysiologic study (EPS) in the patients with unexplained syncope, the electrophysiologic findings of 268 patients with unexplained syncope despite a complete clinical evaluation were analyzed. Results showed positive EPS finding was 38% in total patients and 50% in the patients aged > 70 years. With increasing age, the diagnostic yield of EPS also increased. No significant differences of complication rate were found among the different age groups. It was concluded that EPS have high diagnostic value in the patients with unexplained syncope. Its complications are few and mild. EPS may be recommended in elderly patients with unexplained syncope. PMID- 14526434 TI - Expression of angiogenic factors in hepatocellular carcinoma after transcatheter arterial chemoembolization. AB - In order to investigate the changes of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) expression in residual hepatocellular carcinoma (HCC) after transcatheter arterial chemoembolization (TACE), the expression levels of VEGF and bFGF expression in specimens surgically removed from 48 HCC patients were detected by immunohistochemical methods, and staining intensity of VEGF and bFGF was assessed by a computer-assisted image-analyzer. Among the 48 patients, 25 underwent partial hepatectomy alone (single operating group), and 23 were subjected to second stage surgical resection after TACE (TACE group). The results showed that the average absorbance value (A) of VEGF was higher in TACE group than that in single operating group (0.152 +/- 0.021 vs 0.131 +/- 0.012, P < 0.01). The Average A of bF-GF in TACE group was 0.127 +/- 0.023, higher than in single operating group (0.111 +/- 0.016, P < 0.05). These results suggested that TACE of HCC can up-regulate the expression of VEGF and bFGF in HCC tissues possibly due to anoxia and ischemia. PMID- 14526435 TI - Effects of estradiol and tamoxifen on proliferation of human breast cancer cells and human endometrial cells. AB - The effects of estradiol and tamoxifen on the proliferation of estrogen receptor positive cells and the relationship between the tamoxifen tolerance and cell origin were investigated. The tissues of human endometrium and breast cancer were randomly selected following dissection for primary cell culture. After the breast cancer cells and endometrial cells were treated with 1 x 10(-8) mol/L estradiol and/or 1 x 10(-6) tamoxifen, 3H-labelled thymine nucleotide was used to trace the kinetics of cell proliferation. There was no significant difference in the inhibition on the human endometrial cells between tamoxifen-treated group (6.3%) and control group (6.4%), but tamoxifen could significantly inhibit the proliferation of the human breast cancer cells (45.84%) as compared with control group (52.72%). Moreover, tamoxifen could significantly stimulate the proliferation of tamoxifen resistant breast cancer cells (9.64%) as compared with control group (6.32%). Estradiol could significantly stimulate the proliferation of all the three kinds of cells as compare with control group. The combined use of estradiol and tamoxifen could inhibit the proliferation of the endometrial cells and breast cancer cells as compared with estradiol used alone, but on the tamoxifen resistant breast cancer cells, they could more significantly stimulate the proliferation than E2. It was concluded that E2 could stimulate the proliferation of these three kinds of cells. However, the inhibitive effects of tamoxifen on the proliferation of these cells were dependent on the estradiol. PMID- 14526436 TI - Effect of estrogen on telomerase activity in human breast cancer cells. AB - To investigate the effects of estrogen (E2) on telomerase activity and its mechanism in human breast cancer cells, estrogen receptor positive MCF-7 cells were treated with different concentrations of E2. Telomerase activity was measured by using TRAP-ELISA method, the cell cycle phases analyzed by using flow cytometry, and the expression of Cyclin D1 detected by using immunohistochemistry method. The results showed that telomerase activity levels were increased in MCF 7 cells treated with 10(-8) mol/L E2 during the observed period (P < 0.05), and E2 increased telomerase activity levels in a dose-dependent manner(10(-10)-10(-8) mol/L); Simultaneously, the cell cycle phases of MCF-7 cells treated with 10(-8) mol/L E2 were changed significantly: G0/G1 phase decreased from 60.52% to 50.93%. S phase increased from 29.03% to 30.83%; However, the expression of Cyclin D1 was decreased. It was concluded that estrogen can upregulate telomerase activity of MCF-7 cells, and the effect can be blocked by antiestrogen tamoxifen. Its mechanism may be closely associated with modulation of cell cycle phases. PMID- 14526437 TI - Expression of mouse telomerase catalytic subunit mTERT gene in testis of SD rats and its significance. AB - To study the expression of mTERT gene in the testis of SD rats and its significance, in situ hybridization (ISH) techniques were used to detect the expression of telomerase gene mTERT mRNA in the testis of SD rats. The expression of mTERT was detectable in different-age male SD rats testis. There was a positive correlation between the expression of mTERT and the location of germ cells (spermatogonia, spermatocyte, spermatid). In Sertoli cells, leydig cell and spermatozoa, telomerase mTERT was not detected. Type A spermatogonia expressed the highest level of telomerase mTERT mRNA. Our results suggest that the expression of mTERT gene in the testis of SD rats is of lifetime and coincide with the telomerase activity. PMID- 14526438 TI - Expression of beta-human chorionic gonadotropin genes in renal cell cancer and benign renal disease tissues. AB - To study the expression of beta-human chorionic gonadotropin (beta hCG) genes in renal cell carcinomas (RCC) and benign renal disease tissues, nested reverse transcription-polymerase chain reaction (RT-PCR) and restriction endonuclease analysis were employed to detect the expression of beta hCG genes in 44 cases of RCC tissues and 24 cases of benign renal disease tissues. It was found that 52% RCC samples revealed positive for beta hCG mRNA expression. Positive rate in advanced stage and poorly differentiated RCC was higher, but there was no significant difference. The positive rate of beta hCG mRNA expression was 54% in 24 cases of benign renal tissues, including 3 cases out of 6 polycystic kidneys, 7 cases out of 13 renal atrophies, 2 cases out of 2 oncocytomas and 1 case out of 2 pyonephrotic kidneys. beta 7 was most frequently transcribed subtype gene independent on the histology. These findings suggested beta hCG gene transcription is not only involved in RCC but also in benign renal diseases. PMID- 14526439 TI - Analysis on pathogenesis of 50 cases of bladder proliferative lesions. AB - In order to study the pathogenesis, clinical and pathological characteristics of proliferative lesions of the bladder, 50 cases of proliferative lesions of the bladder from 150 patients with complaints of frequency, urgency, hematuria and dysuria were subjected to cystoscopic biopsy of the suspicious foci in the bladder. In combination with the symptoms, urine and urodynamics, the relationship of proliferative lesions of the bladder to the inflammation and obstruction of the lower urinary tract was analyzed. Of the 50 cases of proliferative bladder lesions, 44 cases (88%) had lower urinary tract infection and 29 (58%) lower urinary tract obstruction. The patients with lower urinary tract obstruction were all complicated with infection. Three cases were associated with transitional cell carcinoma. Malignant cells were detected in 1 case by urinary cytologic examination. Proliferative lesions of the bladder, especially those without other obvious mucosa changes under cystoscopy, are common histological variants of urothelium in the patients with chronic inflammation and obstruction of the lower urinary tract. Chronic inflammation and obstruction of the lower urinary tract might be the causes for proliferative lesions of the bladder. It is suggested that different treatments should be applied according to the scope and histological type of the proliferative lesions. PMID- 14526441 TI - Anterior lumbar intervertebral fusion with artificial bone in place of autologous bone. AB - The feasibility of anterior lumbar intervertebral fusion with artificial bone in place of autogenous bone was investigated. Porous hydroxyapatite (HA)/ZrO2 ceramics loading bone morphogenetic protein (BMP) were implanted after removal of lumbar vertebral disc in rabbits. The adjacent intervertebral discs were also removed by the same way and autogenous illic bone was implanted. SEM observation and biomechanical test were carried out. Compound bone had a bit lower osteoinductive activity than autogenous bone by SEM (Osteoinductive activity of artificial bone in 12 weeks was the same as that of autogenous bone in 9 weeks). Biomechanical test revealed that compound bone had lower anti-pull strength than autogenous bone (P < 0.001), but there was no significant difference in anti-pull strength between compound bone at 12th week and autogenous bone at 9th week (P > 0.05). It was concluded that compound bone could be applied for anterior spinal fusion, especially for those patients who can't use autogenous bone. PMID- 14526440 TI - Experimental study of vascular endothelial growth factor gene therapy for avascular necrosis of the femoral head. AB - To explore a new method for the therapy of the avascular necrosis of the femoral head, the recombinant plasmid pCD-hVEGF165 was mixed with collagen and was implanted in the necrotic femoral head. The expression of vascular endothelial growth factor (VEGF) was detected by RNA dot hybridization and immunohistochemical method. The repair of the femoral head was observed by histological method. The results showed that the expression of VEGF was detectable in the femoral head treated with VEGF gene. Angiogenesis in these femoral heads was more abundant than the control. Bone repairing was augmented in the femoral head treated with VEGF gene. The results suggest that angiogenesis in bone tissue could be augmented by gene transfection of VEGF and bone repairing would be accelerated accordingly. PMID- 14526442 TI - Expression of TGF-beta in region of bone defect repaired by collagen/nano-beta tricalcium phosphate composite artificial bone. AB - The distribution and function of transforming growth factor-beta (TGF-beta) in the region of bone defect repaired by collagen/nano-beta-tricalcium phosphate composite artificial bone (Co/N-TCP) and the ability of Co/N-TCP recruiting osteoblasts to precipitate the repair of bone defect were investigated. Twenty four domestic rabbits were operated on bilateral cranial bone to create an experimental bone defect of 8.0 mm in diameter through the whole bone. On the left, Co/N-TCP was implanted as experimental group, but on the right, Co/TCP was implanted as control group. At 2nd, 4th, 8th, 12th week after operation, all animals were sacrificed and the implanted materials with surrounding bone were taken out. Immunohistochemical staining was performed for TGF-beta assay by avidin-biotin complex method (SABC). Simultaneously, TGF-beta was quantitatively analyzed by HPIAS-1000 imaging analysis system. The immunohistochemical staining for TGF-beta revealed that osteoblasts and immature osteocytes highly expressed TGF-beta. Diffused TGF-beta positive staining particles appeared in the mesenchymal and fibrous-tissue. There was no significant difference in the TGF beta positive staining between two groups in the medial region to original osseous beds at different time points (P > 0.05). However, in distal original osseous bed of the defected region, the positive expression of TGF-beta in the Co/N-TCP group was significantly stronger than in the control group (P < 0.05 or 0.01). The Co/N-TCP has good bioactivities and ability of stimulating and conducting TGF-beta to aggregate and precipitate the healing of bone defect. PMID- 14526443 TI - Effect of hypertension on hearing function, LDH and ChE of the cochlea in older rats. AB - The relationship between the hypertension and the aging process of hearing organ was investigated. Twenty Wistar 3-month old rats and 20 Wistar 12-month old rats, 20 spontaneously hypertensive rat stroke-prone (SHRSP) 3-month old rats and 20 SHRSP 12-month old rats free of middle ear infections as observed under otomicroscopy, with normal tympanic membrane and auricle reflex, were selected to be divided into two experimental groups and two control groups respectively. The tail artery blood pressure was measured non-invasively. The threshold of auditory brain-stem response (ABR) was measured by Spirit evoked potential meter. The LDH and ChE staining in the inner ear was performed and the optical density was analyzed by the HPIAS analysis system. The results showed that there was no difference in the ABR thresholds, the activities of LDH and ChE between Wistar 3 month old group and SHRSP 3-month old group (P > 0.05). The mean value of ABR threshold and the activities of LDH and ChE in the Wistar 12-month old group at relevant sections were significantly greater than those in the two 3-month old groups (P < 0.05), whereas the mean value of ABR threshold and the activities of LDH and ChE in the SHRSP 12-month old group at relevant sections were significantly higher than those in the 3-month old control group (P < 0.01). It was concluded that presbycusis existed in the Wistar 12-month old group rats. The glycogenosis and the abnormal secretion of neural transmitter were discerned after hypertension. All the above factors may worsen the aging of the hearing system. PMID- 14526444 TI - Expression of HSP70 in peripheral lymphocytes of the patients with allergic rhinitis. AB - The expression levels of heat shock protein 70 (HSP70) from peripheral lymphocytes of the patients with allergic rhinitis (AR) and the clinical implication were investigated. In the morning, 3 ml of fasting venous blood was taken out. The lymphocytes were isolated by using Ficoll-Hypaque and the expression of HSP70 in the lymphocytes was detected by using Western blot. In the AR patients the HSP70 level (41.49 +/- 15.77 integrated optical density, IOD) were significantly higher than that in the control group (23.89 +/- 10.13 IOD, P < 0.05). Western blot demonstrated that HSP70 bands in AR patients were more intensive than those in the control group. It was concluded that the elevated HSP70 level in peripheral lymphocytes of the AR patients might contribute to the development of AR. PMID- 14526445 TI - Effect of catecholamine on central serous chorioretinopathy. AB - The role of catecholamine (CA) in the pathogenesis and development of macular edema of central serous chorioretinopathy (CSC) was studied, and its relations with visual acuity were investigated. Plasma concentrations of epinephrine (E) and norepinephrine (NE) were determined in 30 consecutive eyes with CSC. Central macular thickness analysis was done by RTA and all the data were compared with normal eyes and analyzed with SAS software package. Plasma concentrations of E and NE were increased to (569 +/- 123) ng/L and (721 +/- 104) ng/L respectively in active CSC patients, significantly higher than those in normal subjects (P < 0.01), and decreased to normal in convalescent stage. RTA analysis revealed that the retinal thickness of CSC patients was increased at active and recovery stage as compared with normal subjects; and the plasma concentration of E was significantly correlated with central macular thickness (t = 2.173, P < 0.05). Also, central macular thickness measured by RTA was significantly correlated with the visual acuity (r = -0.8046, P < 0.001) in CSC eyes. RTA analysis might be useful to quantitatively detect and evaluate prognosis in CSC patients. The plasma concentration of E, which was highly correlated with macular edema, might play an important role in the early damage and the pathogenesis of CSC. PMID- 14526446 TI - The lymphoblastoid cell lines of recurrence condyloma acuminatum patients produce lower level of tumor necrosis factor stimulated with LPS. AB - To study the mechanism of Condyloma acuminatum (CA) recurrence, and the association of CA recurrence with the ability of the host derived lymphoblastoid cell lines (LCL) stimulated by LPS to produce tumor necrosis factor (TNF), EBV transformed B LCL were used as TNF producing cells. The ability of LCL stimulated by LPS to produce TNF was measured by bioassay. The results showed that the LCL from CA patients (including recurrent and non-recurrent CA patients) produced similar level of TNF stimulated by LPS to that of normal controls (29.54% +/- 11.28% vs 34.31% +/- 11.46%, P = 0.1498). The LCL of CA recurrent patients produced significantly lower amount of TNF than that of non-recurrent CA patients (23.72% +/- 7.41% vs 37.33% +/- 11.10%, P = 0.0032). Compared with the normal controls, CA recurrent patients showed a decreased ability to produce TNF (23.72% +/- 7.41 vs 34.31% +/- 11.46, P = 0.0054), whereas CA non recurrent patients had the similar ability to the controls (37.33% +/- 11.10 vs 34.31% +/- 11.46, P = 0.4914). It was concluded that the onset of CA was not relevant to the individual's ability to produce TNF. But the recurrence of CA was associated with the ability to produce TNF. It was also indicated that the TNF involved cellular immunity might play an important role in the clearance of the residual HPV by the host after treatment. PMID- 14526448 TI - Extranodal imaging manifestations of non-Hodgkin's lymphoma. AB - A series of imaging features of extranodal, multi-systemic involvements in Non Hodgkin's lymphoma (NHL) were investigated. The clinical data and imaging findings of 16 patients with pathologically proved NHL were retrospectively analyzed. The related literatures were reviewed. Of the 16 cases of NHL, skeletal involvement was found in 4, nasal cavity and nasal sinuses were involved in 4, too. Lesion in the thorax was seen in 3 patients, hepatic involvement occurred in one case, cerebral ventricle was affected in 3 cases, mesentery was involved in one case. Even though extranodal involvement of NHL exhibited extremely variable patterns, there were some relatively typical imaging findings. Emphasized in this report were the relatively specific imaging manifestations of different systems, which may mimic infectious or other neoplasms of different sites. The importance of imaging studies lies in the availability for diagnosis, staging and follow-up of NHL. Combined with the clinical and other related information, the diagnostic accuracy can be further improved, thus, providing reliable evidence in guiding clinical management. PMID- 14526447 TI - Therapeutic effect of propofol in the treatment of endotoxin-induced shock in rats. AB - To assess the potential therapeutic effect of propofol in the treatment of endotoxemia, 76 rats were randomly assigned to 5 groups: control group(A), endotoxemic group(B), pre-treatment group(C), simultaneous treatment group(D) and post-treatment group(E). Five h after endotoxin injection, PO2, pH, MAP, plasma concentrations of Nitrite/nitrate (NO2-/NO3-) and mortality rates were assessed in each group. After the rats were sacrificed, lung tissue was sampled to measure myeloperoxidase (MPO) activity and tumor necrosis factor (TNF)-alpha contents. It was found that endotoxin injection produced progressive hypotension, metabolic acidosis, and a large increase in the plasma NO2-/NO3- concentrations and increased mortality rates in 5 h. Endotoxin injection significantly increased MPO activity and TNF-alpha contents in lung tissue (P < 0.01 or P < 0.05). These changes response to endotoxin were significantly attenuated in the groups B, C and D. But these beneficial effects were blunted in the group E. The results suggest that propofol administration may offer advantages in endotoxemia. PMID- 14526449 TI - [Telemetric system for measuring the pressure of the tongue on the palate]. AB - The development of the upper jaw during growth is influenced by the function and position of the tongue and perioral soft tissues, and the pressures exerted by them. Accurate determination of the forces exerted by the tongue would provide relevant information about this influence. To date, our ability to obtain continuous recordings of the tongue pressure during certain functions is limited. In this paper, an easy-to-employ and accurate telemetric system for such functional measurements is presented. The system, consisting of four piezoresistive pressure sensors, a microcontroller, a telemetric module and batteries, is integrated within a removable orthodontic plate and transmits the measured data out of the oral cavity to a receiver. PMID- 14526450 TI - [Telemanipulatory application of a magnetic coupler on the beating heart with the daVinci surgical system]. AB - The construction of a coronary anastomosis on the beating heart under totally endoscopic conditions is technically demanding. In this study the potential benefits of an endoscopic magnetic vascular coupler (MVP, Ventrica, Inc, Fremont, CA) designed to facilitate construction of a coronary anastomosis with the help of the daVinci telemanipulator (Intuitive Surgical Inc., Sunnyvale, CA) were evaluated in a totally endoscopic coronary arterial bypass (TECAB) operation on the beating heart in eight dogs. The telemanipulated instruments were used to guide and place the endoscopic MVP-application platform (prototype). All animals underwent angiography, and gross inspection of the anastomotic site was done after excision of the hearts. The procedure was accomplished in 169 minutes (155 190). With the exception of one premature deployment, all MVP-anastomoses were accomplished in 3 minutes (1-28). The following adverse events were encountered: Bleeding from the right ventricle caused by occlusion tape (1), anastomotic leakage upon reperfusion requiring repair stitches (2), anastomotic occlusion due to a thrombus (1). All but one animal that died on reperfusion despite a patent graft and anastomosis, survived the procedure. Overall patency was 7 out of 8. The combination of telemanipulator technology allowing increased manipulation dexterity in a total endoscopic environment and the effective and time saving magnetic technique for anastomotic coupling has the potential to facilitate TECAB on the beating heart. PMID- 14526451 TI - Ensemble averaging and multiple statistical testing of EMG activities of cyclically repeated body motions. A tool for muscle function analysis in experimental and clinical orthopaedics. AB - Coupled with suitable computerized signal recording and processing methods surface electromyography can be a powerful tool for the analysis of muscle activity in specific body movements. It can be used for this purpose in experimental and in clinical diagnostic orthopaedics as well as in physiotherapy. We describe in this paper a motion analysis system comprising this feature. It has been employed for the diagnosis of the basic angular kinematics and muscle function in human gait and other cyclically repeatable movements of the human locomotive system. Changes in the temporal characteristics of the movements and the muscle activity due to changed physical or experimental conditions can be systematically investigated this way. Such changes can be the result of surgical and/or conservative orthopaedic therapy, a long term physiotherapeutic program, or modified walking conditions as in experimental orthopaedics. They are displayed and validated by signal ensemble averaging and subsequent multiple statistical testing (e.g. by a suitably adapted Bonferroni criterion). The efficiency of the system is demonstrated by an exemplary gait analysis of selected kinematic and muscular effects caused by an experimental simulation of a leg length inequality. PMID- 14526452 TI - Influence of different culture solutions on osteoblastic differentiation in cord blood and bone marrow derived progenitor cells. AB - Mesenchymal progenitor cells derived from cord blood (unrestringated somatic stem cells, USSC) and bone marrow (mesenchymal stem cells, MSC) are able to differentiate under defined culture conditions into at least bone, cartilage, adipose and muscle cells in vitro. The culture media and other in vitro conditions influence the osteogenic differentiation potency of both cell types. To increase and expand the number of osteoblasts in vitro an optimization of culture conditions is required. The aim of this study was to evaluate different culture media toward their osteogenic promoting capacity on human USSCs and MSCs in vitro. Immunohistochemical stainings against osteonectin (ON), osteopontin (OP) served as markers for an osteoblastic differentiation. Cellular morphology was analysed by light microscopy technique. We found significant differences between bone marrow and cord blood derived stem cells towards an osteoblastic differentiation. Considering the number of osteoblasts MesenCult seems to have advantages in bone marrow progenitor cells, whereas low glucose DMEM and HAMS-F12 promoted an osteoblastic differentiation in cord blood derived cells more than other tested media. PMID- 14526453 TI - [Cost-benefit analysis in patients with sleep-related breathing disorders - diagnosis and ncpap therapy during medical rehabilitation]. AB - In a multi-centre study, 745 patients undergoing internal medical rehabilitation (for hypertension, coronary heart disease, gastrointestinal and respiratory diseases) were investigated. The health economic benefit was evaluated during the 3 weeks of medical rehabilitation, during which a sleep-medical diagnostic work up and treatment were applied. Ambulatory screening for sleep-related breathing disorders was carried out in all patients. In positive cases (Apnoea-Hypopnoea Index > or = 10) transfer to our sleep lab was recommended. 103 patients were found to be positive, of whom 47 attended the lab; 23 of these accepted nCPAP therapy, while 24 did not. The costs of the additional diagnosis and treatment were considered incremental costs--and the benefit identified as the decrease in days off work as revealed by a comparison of the year before with the year after rehabilitation. Days off work decreased by 38.4 days in the treated group, and increased by 25.4 days in the untreated group. The results were extrapolated to all patients in internal medical rehabilitation in Germany, and a cost-benefit analysis showed that the benefit of expanding the additional investigation to cover all patients would far exceed the incremental costs in the first year after rehabilitation by 58.26 Mio [symbol: see text] and in the second year by 81.15 Mio. [symbol: see text]. PMID- 14526455 TI - [Two classes of medicine: what are the consequences of treatment advances with new recombinant proteins?]. PMID- 14526454 TI - [Design, construction and modularity of pressure-fit acetabular cups]. AB - To enable a comparison of different pressfit acetabular cups objective criteria are essential. The aim of this study is to describe the design features of this type of cup and to analyse currently available cups. 30 implants were systematically measured and analysed. The mean surface roughness (Ra) was determined and configurations established with the light section technique. For further evaluation the cups were transversely sectioned. The cups are made of pure titanium, titanium alloy or polyethylene coated with titanium. Five implants take the form of monoblocks. The configuration is predominately (n = 25) flattened spherical. The size of eight cups corresponds to the outer diameter, 19 cups have a larger outer diameter (overdimensioning), 3 cups have a smaller outer diameter (underdimensioning). The magnitude of overdimensioning is, on average, 1.9%. 9 cups are provided with plugs, hollow cylinders, fins or rings as outer stabilizers. Surface roughness achieved with corundum blasting is 6.8 microns. Titanium porous-coated implants have a surface roughness of 21-32 microns. 24 cups have polyethylene inserts, most of which are snap-fixed with equatorial lips. For 16 cups, full-ceramic inserts are available. 4 cups have a metal insert. Titanium implants with structured or HAC-coated surfaces have become the accepted standard for cementless acetabular cup implantation. Together with ceramic, metal, or modified polyethylene inserts they meet the requirement for permanent osteo-integrative stability. PMID- 14526456 TI - [The pathologic basis: pathogenesis of rheumatoid arthritis]. PMID- 14526457 TI - [Kineret, Enbrel, Remicade and company. Recombinant drugs in rheumatoid arthritis]. PMID- 14526458 TI - [Problems with drug stability. Galenic formulation of recombinant drugs]. PMID- 14526459 TI - [An important step toward success. The safety profile of recombinant antirheumatics]. PMID- 14526460 TI - [Assessment of the studies. Important recombinant agents in treatment of rheumatoid arthritis]. PMID- 14526461 TI - [When do we use which drugs? Procedures for the treatment of rheumatoid arthritis]. PMID- 14526462 TI - [Questions and answers. Rheumatic patients under treatment with recombinant drugs]. PMID- 14526463 TI - [Cost effectiveness of pharmaceutical innovations. Health economic aspects of rheumatoid arthritis treatment with TNF-alpha antagonists]. PMID- 14526464 TI - [Antimicrobial susceptibility testing of bacteria isolated from animals: methods for in-vitro susceptibility testing and their suitability with regard to the generation of the most useful data for therapeutic applications]. AB - In-vitro susceptibility testing provides valuable informations for choosing the most suitable antimicrobial agent for the control of bacterial infections in animals. Different diffusion and dilution methods, as conducted according to various approved performance standards, can be used to determine the in-vitro susceptibility of bacterial pathogens. In the present article, problems are discussed which arise from the use of different methods and the difficulty to interpret such results. While most approved performance standards were designed for testing of bacteria from human sources, the NCCLS document M31-A2 exclusively focusses on susceptibility testing of bacteria isolated from animals and--in contrast to all other standards--includes veterinary specific breakpoints for a number of antimicrobial agents used in veterinary medicine. Therefore, performance of in-vitro susceptibility testing of veterinary pathogens should follow the recommendations given in the NCCLS document M31-A2. The microdilution method is recommended as the method of choice for susceptibility testing. The result of a microdilution test is given as the minimum inhibitory concentration (MIC). This value provides a quantitative result which precisely indicates the degree of susceptibility of the tested bacterial strain and in return gives the veterinarian a clear guidance whether therapeutic intervention with the antibiotic in question will be successful. PMID- 14526465 TI - Evaluation of the oral immunisation of wild boar against classical swine fever in Baden-Wurttemberg. AB - The oral immunisation of wild boar against classical swine fever (CSF) in Baden Wurttemberg is described and evaluated. The bait vaccine based on the CSF virus (CSFV) strain "C" proved to be safe in wild boar of all age classes. The modified immunisation procedure consisting of three double vaccinations per year was very effective. CSFV was not detected beyond the second immunisation campaign. The average rate of seropositive wild boar diagnosed over all immunisation periods was 49.2%. The seroprevalence rate increased significantly during the first year of immunisation and reached its maximum after the third vaccination period with 72% antibody positive animals. The higher percentage of seropositive young boars in this field trial compared to the seroprevalence rates in this age class in other field trials in Germany may be attributed to the new vaccination scheme. Factors that may be responsible for the decreased herd immunity after the fourth or sixth immunisation period are discussed. PMID- 14526466 TI - [Canine brucellosis--a case report]. AB - Brucella (B.) canis was isolated from ejaculate of a 4-year old Korthals-Griffon male dog after occurrence of epididymitis and orchitis. Despite several trials of therapy with different antibiotics relapes occurred, with B. canis being isolated from ejaculate, blood and urine samples, respectively. Bacteriological examinations were added by serological testing over a period of about 1.5 years. During the study SAT serum titre steadily dropped from 1:200 to 1:50. By CFT, B. canis antibodies were detectable at the beginning with a titre of 1:320 and to the end of the study with titres between 1:80 and 1:160. PMID- 14526467 TI - [Establishment and use of infectious bacterial artificial chromosome (BAC) DNA clones of animal herpesviruses]. AB - Since the first description of the cloning und mutagenesis of a herpesviral genome in 1997, a number of herpesviral DNA's have been cloned as bacterial artificial chromosomes (BACs). Herpesvirus BAC's are established by introduction of a mini F origin of replication into the viral genome, which can as such be maintained und manipulated in Escherichia coli as a single copy extrachromosomal DNA. Herpesvirus DNA's cloned as BAC's are accessible to the DNA repair und mutagenesis apparatus encoded by E. coli, which can be exploited for the generation und subsequent analysis of virus mutants. One of the advantages of BAC cloning und mutagenesis is that it is independent of virus growth in cultured cells, thereby minimizing the risk of compensatory mutations. Here we summarize BAC's that have been established for Herpesviruses of livestock, among them Bovine Herpesvirus Type 1 (BHV-1), Equine Herpesvirus Type 1 (EHV-1), Marek's Disease Virus (MDV), Pseudorabies Virus (PRV). Special emphasis is put on the exploitation of herpesviral BAC's for the analysis of virus replication und vaccine developments. PMID- 14526468 TI - [Avian pathogenic Escherichia coli (APEC)]. AB - Infections with avian pathogenic Escherichia coli (APEC) cause colibacillosis, an acute and mostly systemic disease resulting in significant economic losses in poultry industry worldwide. Avian colibacillosis is a complex syndrome characterized by multiple organ lesions with airsacculitis and associated pericarditis, perihepatitis and peritonitis being most typical. Environmental factors as well as the constitution of poultry or initial viral infections influence the outcome of APEC-infections. However, several challenge experiments in chickens proofed the role of virulent APEC strains as the single aetiological agent. Currently serotypes O1:K1, O2:K1 and O78:K80 are recognized as the most prevalent, however the number of published serotypes is increasing. In addition, single APEC isolates vary profoundly in virulence, and knowledge about the molecular basis of this variability is still scarce. Known virulence factors of APEC are adhesins (F1- and P-fimbriae), iron acquisition systems (aerobactin and yersiniabactin), hemolysins (hemolysinE and temperaturesensitive hemagglutinin), resistance to the bactericidal effects of serum and phagocytosis (outer membrane protein, iss protein, lipopolysaccharide, K/1)-capsule and colilcin production) as well as toxins and cytotoxins (heat stable toxin, cyto-/verotoxin and flagella toxin). Esperimental studies have shown that the respiratory tract, principally the gas-exchange region of the lung and the interstitium of the air sacs are the most important sites of entry for avian pathogenic E. coli. APEC strains adhere to the epithelial cells of air sacs presumably through F1-fimbriae. After colonization and multiplication the bacteria enter the bloodstream, and the temperature-sensitive hemagglutinin (tsh) seems to be important int his step. After invading the bloodstream APEC cause a septicemia resulting in massive lesins in multiple internal organs and in sudden death of the birds. The ability of the bacteria to acquire iron and the resistance to the bactericidal effects of serum, predominantly conferred by the increased serum survival (iss)--protein, enables APEC to multiply quickly in their hosts. Iss is regarded a specific genetic marker for avian pathogenic E. colistrains. A critical review of the literature published so far on APEC reveals, that these pathotypes are not defined appropriately. This findings urge investigations on the population structure of APEC, enabling the establishment of appropriate diagnostic tools and avoiding the obsolete use of serotyping for APEC diagnosis. So far more than 20 APEC strains have been investigated in animal experiments, explaining contrary published results. Thus, the lack of knowledge in pathogenicity and in immunity of APEC infections urges further experimental studies. As APEC share not only identical serotypes with human pathogens but also specific virulence factors, their zoonotic potential is under consideration. PMID- 14526469 TI - Isolation of Vibrio vulnificus and atypical Vibrio from surface water of the Baltic Sea in Germany. AB - From 1995 to 1997 several defined species like V. alginolyticus, V. anguillarum, V. cholerae (non O1 and non O139), V. mimicus, V. parahaemolyticus and V. vulnificus were isolated during a survey to determine the presence of V. vulnificus in the brackish water of the Baltic Sea in Germany. Moreover atypical Vibrio isolates were detected. Four isolates belonging to a group of atypical Vibrio and possibly representing a new species in the genus Vibrio were characterized in detail. All four strains were isolated from surface costal waters. Based on 16S rDNA sequence analysis they showed the highest relatedness to the species V. navarrensis and V. vulnificus. The strains did not harbor the species specific hemolysin gene vvhA from V. vulnificus as shown by PCR and hybridization experiments. Moreover, they differed in at least two biochemical parameters tested from the hitherto described Vibrio species. All these strains induced hemolysis on washed blood agar dishes and showed phase variations on Luria Bertani agar dishes. Because of the similarity to the eel pathogen V. vulnificus, we infected eels with one of the four atypical strains (CH-291), but no pathogenicity for eels could be detected. Furthermore, Vero cell tests with supernatants of bacterial cultures did not reveal secreted Vero cell cytotoxic compounds. This indicates a nonpathogenic nature of these strains. PMID- 14526470 TI - [Phenotypic and genotypic methods for epidemiological typing of veterinary important bacterial pathogens of the genera Staphylococcus, Salmonella, and Pasteurella]. AB - Molecular typing methods are capable of providing detailed strain characteristics which are commonly far beyond the capacities of phenotypic typing methods. Such molecular-based characteristics have proved to be very helpful in epidemiological studies of bacterial pathogens. The primary criteria that all typing methods should fulfill include (1) the typeability of the strains in question, (2) the reproducibility of the results, and (3) a high discriminatory power. In general, molecular typing methods can be differentiated with regard to their use in methods that can be applied to virtually all bacteria (e.g. plasmid profiling, ribotyping, macrorestriction analysis) and methods which can only be used for typing of certain bacterial genera or species (e.g. IS200 typing of certain Salmonella enterica subsp. enterica serovars, or coa-PCR of coagulase-positive staphylococci). In the present review, various phenotypic and molecular methods for the epidemiological typing of bacteria of the genera Staphylococcus, Salmonella, and Pasteurella are described and their advantages/disadvantages- also with regard to the fulfillment of the above-mentioned primary criteria--are critically assessed. PMID- 14526471 TI - Serosurvey for antibodies against Salmonella species in free-ranging moose (Alces alces) from Norway. AB - An indirect ELISA was developed as a tool for surveillance of antibodies against Salmonella sp. in free-ranging moose (Alces alces) in Norway. Serum samples from 303 clinically healthy moose sampled between 1993-2000 were examined. Anti Salmonella antibodies were detected in samples from 6 individuals (1.98%). This is the first evidence of Salmonella-seropositive free-ranging moose. Possible sources and transmission routes of Salmonella comprising environment, wildlife and man are discussed. PMID- 14526472 TI - Investigation on the clinical efficacy and safety of meloxicam (Metacam) in the treatment of non-infectious locomotor disorders in pigs. AB - The clinical efficacy and safety of meloxicam (Metacam 20 mg/ml) in the treatment of non-infectious locomotor disorders in pigs was investigated in a randomised double-blind, placebo controlled, multi-centre field study. A total of 220 pigs were examined, 211 pigs were suitable for evaluation. Treatment was performed on Day 1 with meloxicam (0.4 mg meloxicam/kg) or placebo by intramuscular injection. If necessary, treatment was optionally repeated on Day 2. Clinical examinations were conducted daily from Day 1 (immediately prior to initiation of therapy) to Day 4. The primary parameter, mean "Clinical Lameness Score" (CLS, a sum of the scores of "Lameness at Rest" and "Lameness at Walk"; range 2 to 11) improved from 6.8 and 6.3 on Day 1, to 3.5 and 4.7 on Day 4 in the meloxicam and placebo groups respectively (p < 0.001). At the final examination mean changes from baseline for CLS (Day 1) were 3.25 for meloxicam treated animals and 1.7 for placebo treated animals (p < 0.001). Behaviour score and feed intake improved during the study period with statistically significant differences in favour of meloxicam at all time points after initiation of therapy. Significantly fewer pigs received a second treatment in the meloxicam group than in the placebo group, 46% versus 73% (p < 0.001). A 'very good' or 'good' clinical efficacy assessment was recorded in 83% of the meloxicam cases compared to 42% of the placebo controls at the final examination (p < 0.001). No adverse events were reported due to the use of meloxicam. Furthermore safety of meloxicam in pregnant sows was demonstrated. It is concluded that intramuscular injection of meloxicam (Metacam) at a dosage of 0.4 mg/kg is efficacious and safe for the treatment of non-infectious locomotor disorders in pigs. PMID- 14526473 TI - [A comparison of corneal sensitivity between healthy cats and cats with corneal sequestra]. AB - In order to establish reference values for corneal sensitivity in ophthalmologically healthy persians (n = 40) and domestic short hair cats (n = 60) a prospective study was conducted. Furthermore corneal sensitivity in 48 cats with a corneal sequestrum was measured. Corneal sensitivity was recorded with the help of the aesthesiometer according to Cochet and Bonnet in five different corneal locations (central, nasal, dorsal, temporal, and ventral). The sensitivity for the central corneal region was recorded as amounting to 3.58 +/- 0.56 cm in ophthalmologically healthy domestic short hair cats and to 2.97 +/- 0.58 cm in healthy persian cats. The sensitivity of the central corneal area of a cat with a corneal sequester only amounts to 2.03 +/- 0.53 cm. Between the diseased and the healthy eyes no statistical difference could be demonstrated for any of the measured corneal locations. The sensitivity of the peripheral corneal locations is significantly lower than that of the central corneal region in all three groups examined. PMID- 14526474 TI - [Methodical influence on selected parameters of the acid-base equilibrium in urine samples from dairy cows]. AB - Estimation of net acid base excretion (NABE) in urine samples of dairy cows has often been reported in the veterinary literature. NABE has been proved to be meaningful in estimation of disturbances in the cows acid base equilibrium. In order to examine the meaningfulness of this method different factors with possible influences on NABE were tested. Estimation of simple and fractionated NABE lead to comparable results. Whereas implementation of fractionated NABE is linked with higher expense, simple NABE gives information at lower expense. After four days storing of urine samples in the refrigerator the concentration of NH4+ increased significantly. In view of using this parameter to estimate the acid metabolic situation this may lead to false information. Therefore urine samples should be examined at least within three days after collecting and refrigerated storing. Sediment distribution significantly influenced all urine parameters. Above all the shift of the values into direction of the reference limit may result in the assessment of individual cows as risk group. Homogeneous mixing of the urine samples must be carried out before NSBA-determination. A higher air portion in the sample containers lead to a significant increase of the pH in urine samples. To prevent influences of the air on urine samples, sample containers should be filled completely. PMID- 14526475 TI - [Computertomographic characteristics of primary brain tumors in dogs and cats]. AB - Since computed tomography (CT) was introduced to veterinary medicine in the beginnings of the 80s, the CT of the skull has mainly been used for the diagnosis of brain tumors. The present study includes 18 dogs and five cats, all patients of the Clinic of Small Animals of the School of Veterinary Medicine Hannover. After a CT of their skull was made, a primary intracranial neoplasm was confirmed by a pathomorphological and -histogical examination post mortem. Four of the five cats had a meningioma, one cat an astrocytoma. With five cases the meningioma also predominated in the dogs, followed by primary lymphoma with four cases. An astrocytoma, an oligodendroglioma and a choroid plexus papilloma were diagnosed respectively in two dogs. A pituitary gland tumor, a germ cell tumor and a medulloblastoma represented individual cases. Based on the pathomorphologically and -histologically confirmed diagnosis, characteristics of these primary brain neoplasms can be found in their CT image. However, changes are not pathognomonic for a certain type of tumor; a definitive diagnosis cannot be made from the CT image. PMID- 14526476 TI - [Treatment of condylomata acuminata with the neodymium laser]. AB - The effectiveness of neodymium laser therapy was presented in the treatment of urogenital condyloma acuminata in 55 patients (51 men and 4 women). The patients were followed up 14 days, 4 weeks, 3 months and 6 months after the laser treatment. The main localization of condyloma acuminata was foreskin and glans penis. The cure rate after the 1st session of treatment was 76%, 8.5% of patients required 3 sessions of the laser therapy. After 6 months 7 patients discontinued the follow-up, 89.6% (43 patients) were cured, 2 had the scars of foreskin and 3 suffered from recurrences. Neodymium laser treatment of urogenital condyloma is a safe treatment and no major side effects were encountered. PMID- 14526477 TI - [Occurrence of risk factors for ischemic heart disease among women--health service workers]. AB - Ischaemic Heart Disease (IHD) is still an essential health problem in many countries. In Poland it is one of the leading causes of cardiac deaths, while in western countries and in the USA mortality rate due to heart diseases decreases. The prevention of coronary vessels diseases is a long and a hard process. There is a need to start an early prophylactic action. The programs of primary and secondary prevention should be directed to as many people as possible. In individual strategy any action should be based on the assessment of the degree of complete risk with regard to every single person. Directives and standards elaborated by the Committee of Prophylaxis are helpful. In this research the group of 105 health service female workers were prepared to carry out further preventive searches towards the IHD danger. The following factors were taken into account: the level of total cholesterol (LDL, HDL and triglycerides level), hypertension, cigarette smoking, overweight and obesity, diabetes. The factors mentioned above defined as global risk elements were examined among three age groups: 35-39 years, 40-45 years, 46-50 years. Coronary hazard was established in accordance with the Card of Prophylaxis IHD of Polish Cardiological Society. The main purpose of this research was to identify the IHD risk factors as well as to evaluate its danger among women. PMID- 14526478 TI - [EEG examination in patients with systemic lupus erythematosus]. AB - We examined a group of 83 patients (81 females and 2 males, aged 19-77) diagnosed as systemic lupus erythematosus (SLE), with the mean duration of the disease of seven years. All patients underwent neurological examination and EEG. We found pathological EEG in 41 cases. The most frequent abnormalities were paroxysmal focal changes (22 subjects) and abnormalities in background activity (19 cases). In the studied group only six patients had epileptic attacks so it is an evident disproportion between the number of patients with epilepsy and the number of paroxysmal changes in EEG. These changes need a special control because of the possibility of epileptic evolution in the future. Patients without neurological deficits also had abnormalities in EEG suggesting the presence of subclinical involvement of central nervous system in SLE. In our group we had much lower incidence of epilepsy in SLE than described in literature. It is probably connected with improving diagnostics and therapy of SLE. PMID- 14526479 TI - [Preliminary results of monotherapy with thalidomide in recurrent and treatment resistant cases of multiple myeloma]. AB - Thalidomide is an antiangiogenic drug, but its mechanism of action is not well known and demands further studies. The recent literature suggests that thalidomide is an effective drug in multiple myeloma patients. The objective of the study was to estimate the efficacy of thalidomide monotherapy in the treatment of refractory and relapsed cases of multiple myeloma. We treated with thalidomide 17 patients (12 males, 5 females), average age 51 (range 42-73 years), mean time since diagnosis to the start of thalidomide treatment was 24 months (range 5-48). All patients revealed the features of progressive disease. The mean number of prior chemotherapy schemes was 2. Three out of 17 patients received high dose chemotherapy followed by autologous stem cell transplantation. Thalidomide was administered as monotherapy at a dose of 200 mg (n = 8), 300 mg (n = 1) and 400 mg (n = 8). The mean time of drug intake was 3 months (1-12). The criteria of clinical response were decline of paraprotein at least 25%, 50% and 75% in comparison to value before the treatment. In 5 cases (33%) 25% reduction of paraprotein was observed, 1 patient achieved 50% decline. In the responder group a tendency to decrease marrow plasmocytosis, total serum protein, beta M-2 and LDH was noticed. The good tolerance of the drug, especially in lower doses, and lack of myelosuppression effect allows to expect, that the combination of thalidomide with other cytostatic drugs will improve the efficacy in patients with refractory or relapsed myeloma. PMID- 14526480 TI - [Satisfaction of need for health services among pensioners with reformed health care]. AB - The objective of this study was to determine the demand for health services and to assess the level of satisfaction of these needs in the new, reformed health care system with reference to pensioners and old age pensioners. The survey was carried out over the period of 4 months among patients undergoing a cure in the Municipal Sanatorium in Naleczow. The group consisted of people with chronic diseases, among which cardiovascular system diseases were the leading ones. All the respondents were pension services users. The study results show that patients at older age, chronically ill or disabled, limited the realization of their health needs and did not begin proper treatment, due to difficult access to health services and bad financial situation. The changes introduced in the functioning of health system during the reform period did not trigger the improvement of medical care in the population of pensioners. PMID- 14526481 TI - [Breast neoplasm in men and women--prognosis comparison]. AB - Male breast cancer is a rare disease, therefore our knowledge of the disease is limited. AIM: Comparison of prognosis of male breast cancer and not-otherwise specified (NOS) invasive ductal female breast cancer and invasive lobular female breast cancer. MATERIAL AND METHODS: From 1.01.1997 to 31.03.2000 in Clinical Department of Surgical Oncology, Medical University of Lodz, 3862 patients were operated on breast cancer. There were 24 males (0.6%) and 3838 females (99.4%). Full clinical and pathological data concerning 22 males were collected; these males composed a studied group. Females with invasive lobular breast cancer (65 women) and with invasive ductal NOS breast cancer (474 women) operated on in our Department from 1977 to 1982 composed comparison groups. P-value of = 0.05 was considered as statistically significant. RESULTS: Comparison of survival between males with breast cancer and females with invasive ductal NOS breast cancer: Gehan-Wilcoxon test P = 0.914; Mantel-Cox test P = 0.825; log-rank test P = 0.865; Peto-Peto test P = 0.901. Comparison of survival between males with breast cancer and females with invasive lobular breast cancer: Gehan-Wilcoxon test P = 0.443; Mantel-Cox test P = 0.305; log-rank test P = 0.413; Peto-Peto test P = 0.421. CONCLUSION: Prognosis of male breast cancer is not statistically different from that of female invasive ductal NOS and invasive lobular breast cancer. PMID- 14526482 TI - [Clinical evaluation of hypospadias treatment outcome based on materials from the Pediatric Surgery ward]. AB - The results of surgical treatment of penile hypospadias in 73 boys using D. Browne and Mathieu methods were discussed in the paper. The frequency of urethral fistula formation was used as the criteria of effectiveness. All of the patients were operated on in the Pediatric Surgery Ward in Bydgoszcz. PMID- 14526483 TI - [Electronystagmographic examinations in workers exposed to metallic mercury vapors]. AB - In order to evaluate the vestibular organ electronystagmography was performed in 20 workers employed in the mercury cathode method of potassium chloride electrolysis, and exposed to metallic mercury vapours at air concentrations 0.035 0.05 mg/m3. In 15 (75%) workers a central impairment of the vestibular organ was shown, whereas in 50 healthy controls an abnormal electronystagmogram was found only in 9 (18%) people. Electronystagmography seems to be a useful tool in the assessment of early signs of the central nervous system dysfunction caused by metallic mercury. PMID- 14526484 TI - Pulmonary compensatory indexes in children and adolescents with idiopathic scoliosis I degree. AB - Spirometric and plethysmographic examinations were performed in two groups of children. Group I: 23 children with scoliosis I degree by the Cobb (spinal curvature 17.2 degree) which comprised 12 girls and 11 boys at the age of 8-16 years (the mean age was 11.4 +/- 2.4 yr). The Control Group consisted of 20 children--13 girls and 7 boys at the age of 8-16 years (the mean age was 13.77 +/ 1.72 yr). The measurements were performed using Body-plethysmograph with pneumotachograph Jaeger. Total lung capacity (TLC), forced vital capacity (FVC), forced expiratory volume in 1 sec (FEV1), residual volume (RV), intrathoracic gas volume (ITGV) and at the same time curve indexes, dependence of maximum expiratory flow lung volume, MEF75%VC, MEF50%VC, MEF25%VC were determined. Maximum expiratory flows in relation to vital capacity and total lung capacity were defined as so called compensatory indexes. Slightly decreased values of total lung capacity (TLC) and statistically significantly decreased intrathoracic gas volume (ITGV) as well as residual volume (RV) were found in children with scoliosis in early stage. Forced vital capacity (FVC) and forced expiratory volume in 1 sec (FEV1) were statistically significantly higher in comparison to the control group. Compensatory index MEF50%VC/FVC in children in the early stage of scoliosis was statistically significantly decreased in comparison to the control group (p < 0.02). PMID- 14526485 TI - [Ototoxic drugs. I. Aminoglycoside antibiotics]. AB - The fact that some drugs can affect the hearing function is well known. All aminoglycoside antibiotics have the potential to induce severe and irreversible ototoxicity in the cochlea and vestibular organ. The mechanism causing aminoglycoside-induced hearing loss is still not well established. Nevertheless, today aminoglycosides are the most commonly used antibiotics world wide thanks to their high efficacy and low cost. In this review pharmacological and toxicological properties of aminoglycoside antibiotics are described as well as risk factors and pathophysiology of aminoglycosides ototoxicity. Current hypotheses concerning the mechanisms of aminoglycoside-induced ototoxicity are also discussed. PMID- 14526486 TI - [Stenosis of the carotid arteries]. AB - Atherosclerotic stenosis of carotid arteries is present in approximately 9% of patients above 60 years of age. The stenosis may be silent or may cause cerebral ischaemia--transient or permanent, including severe stroke leading to death. It is estimated that 20-25% of all strokes is caused by carotid artery stenosis in their extracranial course. Ultrasound technique with Doppler recording is an efficient method of carotid arteries stenoses assessment. Asymptomatic carotid artery stenosis treated with acetylsalicylic acid 325 mg/day bears 6-11% risk of stroke in 5 years follow-up. The risk of cerebral ischaemia is significantly higher in persons with symptomatic stenoses. The risk of TIA or stroke recurrence reaches 30-45%. The treatment of carotid arteries stenoses comprises risk factors management and revascularisation procedures. Surgical treatment decreases the risk of stroke by 17%. Surgical endarterectomy perceived as the gold standard of interventional treatment may be efficiently replaced with percutaneous angioplasty with concomitant intravascular stent implantation. PMID- 14526487 TI - [The management of Helicobacter pylori infection in adults--statement of the Polish Working Group and the Maastricht Consensus 2-2000]. AB - Current standards in the management of Helicobacter pylori infection according to the guidelines by European (Maastricht 2-2000 Consensus Report) and Polish Study Group (IX Congress of Polish Gastroenterology Association) were presented in this paper. The comparison analysis was performed as well as the diagnostic methods were described. The indications for Helicobacter pylori infection treatment especially in gastric and duodenal ulcer disease (also complicated by bleeding), gastric precancerous conditions, functional dyspepsia, gastroesophageal reflux disease, non-steroidal anti-inflammatory therapy were discussed. The acceptable schedules of the first and second-line therapy as well as the problem of drug resistance were presented. The special attention was drown to differences between the two sets of guidelines with respect to latest scientific reports. PMID- 14526488 TI - [Treatment of infections associated with Clostridium difficile]. AB - Cases of antibiotic-associated diarrhoea (AAD) associated with toxigenic C. difficile strains more frequently are described recently. In Polish studies diagnosis of AAD or PMC (pseudomembranous colitis) was confirmed by isolation of C. difficile strains in 52% of suspected cases. Strains of C. difficile were isolated also from samples taken from hospital environment (10.5% positive samples). Problem of C. difficile-associated infection is more frequently observed in Poland also because of uncontrolled using of wide spectrum antibiotics. These antibiotics destroy intestinal microflora--"colonization resistance factor". Cases of AAD or PMC were described even after treatment of patients by vancomycin or metronidazol--antibiotics of choice for treatment of C. difficile-associated diseases. Other risk factors as: age, long term hospitalization, previous surgery, colonoscopy are also taken into account, when C. difficile-associated infections are suspected. Different treatment methods of C. difficile-associated infections are reviewed here and discussed in light of recent publications. PMID- 14526489 TI - [Polymorphism of beta 2-adrenergic receptors]. AB - The human beta 2-adrenoceptor is a member of the seven-transmembrane family of receptors. It is expressed in many cell types such as airway smooth muscle cells, neutrophils, eosinophiles, alveolar macrophages and airway epithelial cells. The beta 2-adrenoceptor agonists are the most important group of bronchodilator drugs used in the treatment of asthma. They are classified by their selectivity, affinity for the receptor, potency and pharmacological efficacy. The gene encoding the beta 2-adrenergic receptor is highly polymorphic in the human population. Polymorphism affecting amino acids 16, 27 and 164 are the most common and they have been shown to correlate with some clinical features of asthma, including airways reactivity. They can modulate the behaviour of the beta 2 receptor, altering ligand binding and the characteristics of down-regulation following agonist exposure. The homozygous glycine-16 (Arg-->Gly) variant of the beta 2-adrenoceptor is known to predispose to agonist-induced down-regulation and desensitization, and may play a role in the pathogenesis of asthma severity. The polymorphism at position 27 (Gln-->Glu) is associated with decreased airway responsiveness. The polymorphic variant 164 (Thr-->Ile) shows impaired agonist binding and decreased adenylyl cyclase activity. No convincing evidence has been presented demonstrating a linking of asthma per se with this receptor polymorphism. PMID- 14526490 TI - [Decompensation of brain stem function with sudden death in Arnold-Chiari anomaly]. AB - The authors presented the clinical and anatomopathological description of Arnold Chiari anomaly at 25 years old man, which caused cerebellar manifestation and sudden death. At the base of anatomopathological study the first type of Arnold Chiari anomaly with dislocation of cerebellum tonsils into foramen magnum was recognized. PMID- 14526491 TI - [Progressive myositis ossificans--case report]. AB - Myositis Ossificans Progressiva (MOP) is a rare hereditary condition with ectopic ossifying. This case report demonstrates clinical picture of MOP and possibilities of diagnosis. PMID- 14526492 TI - [Coexistence of Crouzon syndrome with ventricular septal defect]. AB - The authors present the features of Crouzon syndrome and describe presently 17 year-old girl suffering from this syndrome coexisting with congenital heart malformation (ventricular septal defect, successfully surgically treated). We were not able to find any description of such congenital malformations coexistence in the accessible literature. PMID- 14526493 TI - [Epileptic disease and fitness for specific jobs involving driving motor vehicles]. AB - OBJECTIVE: Analysis of the problems the occupational health physician meets in health surveillance of workers with a clinical history of epilepsy, in order to assess fitness for specific jobs requiring driving of motor vehicles. METHODS: A health protocol was used which required acquisition of all information related to job features, work organization, past and current clinical-functional status of epilepsy and any treatment in progress. SUBJECTS: The cases of four workers with clinical history of epilepsy were examined who were judged to be representative both of the most frequent forms of epileptic disease and of jobs requiring driving of motor vehicles. RESULTS: Application of the chosen health protocol allowed us to make balanced assessments of fitness for the different specific jobs performed by the four workers. DISCUSSION: With the use of this protocol it was possible to protect the health and the safety of both the workers concerned and their fellow workers and also, as far as possible, to respect the occupational competence of such workers. We also addressed the problems related to anti-epileptic drug consumption and in itinere road accidents. Lastly, the different medical-legal aspects of the problem were further investigated, analysing national and international legislation with respect to epilepsy and fitness for driving motor vehicles. PMID- 14526494 TI - Occupational risk factors for the sporadic form of Creutzfeldt-Jakob disease. AB - Some case reports among European farmers and a few case-control studies suggested the hypothesis of an increased risk of the sporadic form of CJD (sCJD) associated with livestock farming or work as a butcher. Also, the discovery of the possibility of transmission of the disease via blood or by contact following corneal or dura madre transplant suggested that health occupations might also run higher sCJD risks. However, a meta-analysis of three case-control studies and a multicentre European study did not find any positive association between sCJD and health-related jobs or occupational contact with livestock, such as cattle and sheep, or animal products. To explore possible occupational risk factors for Creutzfeldt-Jakob disease (CJD), we used a publicly available US database including about 6 million deaths in 24 states during 1984-95. Cases were 636 deaths (300 men and 336 women) with CJD (ICD-9 code 046.1) as the underlying cause of death. Controls were 3,180 deaths randomly selected from among those who died from all other diseases except those affecting the central nervous system. CJD cases represented a wide variety of occupations (159) and industries (147). Among occupations and industries, for which previous reports suggested potential exposure to a transmissible spongiform encephalopathy (TSE) agent, the OR for CJD was significantly increased among butchers (OR = 6.8, 95% C.I. 1.5, 30.1, based on 4 cases and 3 controls), and persons working in offices of physicians (OR = 4.6, 95% C.I. 1.2, 17.6 based on 5 cases and 4 controls). Nine other occupations and seven other industries, for which no previous suggestion existed in the literature, also showed significant associations. Overall, our results suggest that occupational exposures are not an important source of sCJD infection. However, as the excess among butchers and some workers in health occupations was consistent with previous reports, more indepth research is warranted to address the hypothesis. PMID- 14526495 TI - Detection methods of the drug-addiction and alcoholism treatment programme of the Spanish National Railway Company (RENFE). AB - BACKGROUND: Social problems in enterprises need to be approached by all groups involved in order to achieve a satisfactory result. With regard to Occupational Health and, specifically, drug-addiction, the groups concerned are Occupational Health Services, Trade Unionists and Managers. OBJECTIVES: To analyse the involvement of the social groups (Occupational Health Service, Management and Trade Unions) in relation to the detection of workers suffering from drug addiction and/or alcoholism. METHODS: During the first eight years of implementation (1990-1997) of the Drug-Addiction and Alcoholism Treatment Programme of the Spanish National Railway Company (RENFE), a total of 477 workers consuming alcohol and drugs were detected and treated under the Programme (1.1% prevalence), 465 men (97.5%) and 12 women (2.5%), with an average age of 39.8 years. RESULTS: The subjects were detected by occupational physicians (55.4%), occupational psychologists (2.9%), managers (11.3%), trade unionists (19.5%), via self-referral (9.4%) and by other means (1.5%). CONCLUSIONS: The results demonstrate the efficiency and the profitability to the company of prevention and treatment interventions and the need for the coordination of managers, trade unions and occupational health services. PMID- 14526496 TI - [Techniques for administering inhalation anesthetic agents, professional exposure, and early neurobehavioral effects]. AB - BACKGROUND: Occupational exposure to high concentrations of anaesthetic gases can cause neurobehavioral effects in operating room personnel. The measures taken to reduce waste gas exposures, including the installation of active scavenging devices and airconditioning systems, are not effective, so that the NIOSH recommendations for maximum exposure are currently unattainable in practice. OBJECTIVES: The aim of the present study was to measure operating room pollution and neurobehavioral functions in a group of anaesthesiologists during open-system and low-flow anaesthesia. METHODS: Environmental concentrations of N2O and isoflurance were measured by an infrared gas analyzer (Bruel & Kjaer) in open system and in low flow anaesthesia. Under the same stress condition, but with different exposure levels to anaesthetic gases, psychomotor vigilance and response speed were evaluated four times with the Reaction Time Test at the beginning and at the end of the first weekday shift and at the beginning and at the end of the last weekday shift. Exclusion criteria were considered excessive alcohol and coffee intake and use of CNS medication. RESULTS: Concentrations of N2O and isoflurane in the operating room were 4.83 ppm and 0.4 ppm respectively, which are lower compared with open systems: 301 ppm and 11.1 ppm respectively. The mean of the Reaction Time was significantly higher (p < 0.01) during work with the open system compared to work in low flow at the end of the first weekday shift and at the end of the last weekday shift. CONCLUSIONS: Low-flow anaesthesia appears to be effective in reducing waste gas exposure: lower flows produced lower values and protect the integrity of neurobehavioral functions. PMID- 14526498 TI - Haemorrhagic pericardial effusion in an asbestos worker. AB - BACKGROUND: Occupational exposure to asbestos may cause pleural and lung disorders and, less frequently, diseases of the peritoneum and pericardium. An exceedingly small number of cases of benign pericardial effusion have been described so far in the medical literature. OBJECTIVES: To report a rare case of haemorrhagic pericardial effusion caused by occupational asbestos exposure in a patient with pre-existent aortic regurgitation, bilateral pleural plaques and no signs of interstitial lung involvement due to asbestosis. METHODS: A thorough clinical and instrumental evaluation (laboratory tests, tuberculin skin test, chest X-rays, transthoracic and transesophageal echocardiography, contrast coronary and aortic angiography, a histological examination of pericardial and pleural surgical specimens) was performed to examine all the known causes of pericardial effusion. RESULTS: The tests performed did not demonstrate any specific cause of pericardial effusion. Surgical assessment three months later, during an aortic valve replacement, showed no signs of aortic dissection or intraparietal hematoma. A nine-year follow up did not reveal any reoccurrence of pericardial effusion. CONCLUSIONS: Asbestos related pericardial effusion is rarely described in the medical literature but must be considered in patients with previous occupational asbestos exposure. There are no specific clinical or pathological aspects indicative of this etiology and the diagnosis remains one of exclusion. A thorough occupational history should be obtained in patients with pericardial effusion of unknown etiology. PMID- 14526497 TI - [Prevalence of extrinsic allergic alveolitis in cattle breeders from the province of Reggio Emilia]. AB - BACKGROUND AND OBJECTIVES: Several new cases of Extrinsic Allergic Alveolitis or Farmer's Lung (FL) in farm workers were reported to Occupational Health Services in the province of Reggio Emilia (Italy). This prompted the Public Health Department to study the prevalence of the disease among milk-cow breeders involved in Parmigiano-Reggiano cheese production: who are the biggest hay users. METHODS: A suitable questionnaire was sent to 1875 farmers in three of the six districts of the province. Half of them (935: 841 males, 94 females) answered; further contacts and medical history research revealed 33 case of "likely FL". Twenty-three (2 females) (10 "missing"), underwent pulmonary function tests, chest X-rays, precipitins tests against Saccharopolyspora Rectivirgula and other fungal antigens and (22 farmers) bronchoalveolar lavage (BAL). RESULTS: According to the "Societa Italiana di Medicina del Lavoro e di Igiene Industriale" diagnostic standards, we found 20 subjects suffering from FL among farmers collecting hay in large cylindrical (round) bales, dried on field (2.6%) and among others still using small (traditional), prismatic bales (0.5%). CONCLUSIONS: The prevalence on the whole exposed population (6000-9000 people) was estimated between 1.5% and 3.0% (90-270 people); no difference was found in FL prevalence between flat and hilly or mountain areas; the method of collecting hay in big "round" bales, dried on field, seems to produce higher frequencies of FL cases if compared with the traditional ones (more frequent in mountain areas). The new hay packing methods, using forced air driers, are suggested as a possible solution. PMID- 14526499 TI - [Protocol for health surveillance of workers exposed to repetitive arm movements]. AB - BACKGROUND: Repetitive movements of the upper limbs at work can cause the onset of musculo-skeletal disorders and therefore an adequate planning of health surveillance is needed. A Work Group on this problem was constituted in Brescia, Italy, following the great interest raised by recent scientific meetings on this topic. OBJECTIVES: The aim of the Group was to prepare a proposal for a health surveillance protocol for the use of Occupational Health Physicians. RESULTS: Health surveillance for the prevention of upper limb disorders must be based on the degree of risk. The risk assessment procedure should be based on the methodology currently available for ergonomic analysis and should also consider the frequency of upper limb disorders in the exposed workers. In case of moderate risk, it is necessary to identify hyper-susceptible individuals, in order to reduce exposure to repetitive movements by means of an adequate task fitness evaluation and suitable health education programmes. In situations of medium-to high risk, a specific programme of health surveillance must be planned in order to identify early disorders and prevent the onset of more severe damage using task fitness evaluation and rehabilitation therapies. The appropriate diagnostic procedure is indicated for this purpose and a classification is proposed to divide the upper limb disorders into two stages, according to the clinical picture: a first acute-subacute stage, which is potentially reversible, and a chronic-subchronic stage, which is non-reversible. Legal reports are required according to the stage of the disease identified and must be supported by an adequate risk assessment. PMID- 14526500 TI - Racial/ethnic diversification in metropolitan and nonmetropolitan population change in the United States: implications for health care provision in rural America. AB - The diversification of the rural population of the United States provides substantial challenges to the current and to future health care systems in rural areas. Because of a variety of historical, discriminatory, and other factors, minority populations have had lower levels of access to health care in rural as well as urban areas and higher rates of both mortality and morbidity than nonminority populations. Although minority health issues have often been seen as primarily urban issues, this article demonstrates that minority population growth has become a major component of total population growth in rural areas in the past several decades (accounting for nearly 62% of the net growth in the nonmetropolitan population of the United States in the 1980s and for nearly 42% in the 1990s), that future US population growth is likely to be largely a product of minority population growth (nearly 89% of US net population growth from 2000 to 2100 is projected to be due to minority population growth), and that the incidence of diseases and disorders in the US population will come to increasingly involve minority populations (by 2050 roughly 43% of all disease/disorder incidences would involve minority population members). The growth of younger minority populations with disproportionately impoverished socioeconomic characteristics will pose challenges for rural areas and health care systems, which also are likely to face health issues created by disproportionately older populations. PMID- 14526501 TI - Quality improvement in critical access hospitals: addressing immunizations prior to discharge. AB - CONTEXT: Many patients hospitalized in critical access hospitals (CAHs) are at high risk for influenza and pneumonia. These hospitalizations may represent a missed opportunity to address immunizations. Addressing these missed immunizations could provide an opportunity for CAHs to gain practical experience in data-driven quality improvement. PURPOSE: To improve documentation and delivery of influenza and pneumococcal immunizations prior to hospital discharge and provide CAHs with quality improvement experience. METHODS: We recruited 17 CAHs in Kansas to participate in a rapid-cycle quality improvement project to address inpatient immunizations. Each hospital identified patient discharges on a monthly basis and abstracted medical records to see if the patient's immunization status had been assessed and if patients had been vaccinated prior to discharge. FINDINGS: Documentation of influenza immunization status improved from 17% of admissions at baseline to 62% at follow-up (P < 0.001). Documentation of pneumococcal immunization status increased from 36% at baseline to 51% at follow up (P < 0.001). Documentation of immunizations was significantly higher among the 8 hospitals that developed standard charting forms for recording immunization status (P < 0.01). Despite improved documentation of immunization status, at remeasurement only 3.4% received an influenza vaccination and 1.3% received a pneumococcal vaccination prior to discharge. CONCLUSIONS: Critical access hospitals can effectively participate in quality improvement activities, but increased involvement of medical staff or standing immunization orders may be needed to improve actual vaccine administration prior to discharge. PMID- 14526502 TI - "Bottoming out?" Among rural African American women who use cocaine. AB - CONTEXT: Studies have explored the spiraling-down effect of losing individual, familial, and social resources among African Americans who use drugs, but there is a gap in knowledge about this "bottoming-out" phenomenon among rural African American women. PURPOSE: The study was conducted to better understand the phenomenon of bottoming out among rural African American women who use cocaine. METHODS: Using an ethnographic approach, researchers drew on multiple qualitative and quantitative data collection methods. Data derived from qualitative interviews, field notes, and demographic profiles describe the phenomenon or lack thereof of bottoming out among 25 southeastern rural African American women who use cocaine. Data collection took place in a rural county of north Florida with a population between 11,000 and 15,000. Twenty-five African American females 18 years or older who used either powder or crack cocaine at the time of enrollment and resided in the rural county participated in the study. FINDINGS: Respondents used 6 major strategies that delayed or prevented them from bottoming out: (1) taking advantage of their social environment and community ties, (2) utilizing various sources of income, (3) accessing family resources, (4) maintaining some degree of discipline over spending for drugs, (5) maintaining routine drug use locations, and (6) renting cheap housing and/or house pooling. CONCLUSION: Unlike studies of similar populations in urban settings, most respondents continued to work after numerous years of drug use, tended not to lose legal custody of their children, maintained a support system of nonusers as well as users over time, and securred food, clothing, and shelter for themselves and in many instances their children using legal or illegal means. PMID- 14526503 TI - A method for identifying places in rural America at risk of not being able to support adequate health services. AB - CONTEXT: Public policymakers and their advisers struggle with the problem of specifying criteria by which health care providers in rural areas are eligible for special consideration in payment policies and for special grant programs. A means of designating places can provide a basis for assistance and can help target public resources for any providers who deliver services in those places. PURPOSE: This paper provides the details underlying a place-based approach to identifying rural areas that are at risk for not being able to provide requisite health services. METHODS: A population size criterion is utilized first to eliminate metropolitan areas and other large agglomerations from consideration. Any territory not included in a place of 3500 or more people, including a 25-mile buffer around that place, is a priori considered to be at risk. All places, including buffers, that have populations between 3500 and 100,000 are further analyzed using population compositional data and principal components analysis. FINDINGS: In 10 states and 24 bordering states selected for developing and testing the method, there were 1907 block groups outside the boundaries of any place with a population of at least 3500. In addition, the analysis suggested that 66 out of 236 places and buffers with populations between 3500 and 100,000 also should be classified as vulnerable. CONCLUSIONS: The results are discussed in regard to how a place-based approach can advance the study of rural health needs. By focusing on the needs of the people residing in a defined area, as determined from the aggregate characteristics of the population, a model is generated that can be used to predict special circumstances confronting any service provider. The public policy implications of the findings are also considered. Special payment policies could be written on the basis of place instead of provider characteristics, and grant programs providing technical assistance could be targeted to places of greatest need. PMID- 14526504 TI - Attitudes of family physicians in Washington state toward physician-assisted suicide. AB - CONTEXT: The topic of physician-assisted suicide is difficult and controversial. With recent laws allowing physicians to assist in a terminally ill patient's suicide under certain circumstances, the debate concerning the appropriate and ethical role for physicians has intensified. PURPOSE: This paper utilizes data from a 1997 survey of family physicians (FPs) in Washington State to test two hypotheses: (1) older respondents will indicate greater opposition to physician assisted suicide than their younger colleagues, and (2) male and rural physicians will have more negative attitudes toward physician-assisted suicide than their female and urban counterparts. METHODS: A questionnaire administered to all active FPs obtained a 68% response rate, with 1074 respondents found to be eligible in this study. A ZIP code system based on generalist Health Service Areas was used to designate those practicing in rural versus urban areas. FINDINGS: One-fourth of the respondents overall indicated support for physician assisted suicide. When asked whether this practice should be legalized, 39% said yes, 44% said no, and 18% indicated that they did not know. Fifty-eight percent of the study sample reported that they would not include physician-assisted suicide in their practices even if it were legal. Responses disaggregated by age groups closely paralleled the group overall. There was a significant pattern of opposition on the part of rural male respondents compared to urban female respondents. Even among those reporting support for physician-assisted suicide, many expressed reluctance about including it in their practices. CONCLUSIONS: These findings highlight the systematic differences in FP attitudes toward one aspect of health care by gender, rural-urban practice location, and other factors. PMID- 14526505 TI - Rural-urban differences in ductal carcinoma in situ as a proxy for mammography use over time. AB - CONTEXT: With emphasis on increasing use of mammography, the rate of ductal carcinoma in situ (DCIS) of the breast has increased dramatically in the United States starting in the early 1980s. It is unclear if rural and urban women have experienced similar increases. PURPOSE: To describe differences in incidence of DCIS between rural and urban women 50 to 69 years of age over time. This may be a proxy indicator of mammography use for the early detection of breast cancer. METHODS: The study population consisted of women 50 to 69 years of age who were diagnosed with DCIS during 1973-1997 and resided in Iowa, Utah, or New Mexico. Data from the Surveillance, Epidemiology, and End Results (SEER) programs in these states were used in the analysis. Location of the woman's residence was classified into urban when she lived in a county considered a Metropolitan Statistical Area, while she was considered rural if she resided elsewhere at the time of her diagnosis. FINDINGS: Increases in rates were different between rural (annual percentage change [APC]: 15.1) and urban women (APC: 34.4). During the 1990s among urban women, the rate of DCIS was still increasing (APC: 3.8), while there was no increase among rural women (APC: 0.2). Differences between rural and urban women within these 3 states were also identified. CONCLUSIONS: The data suggest differences in utilization of mammography between both populations; the increase was lower and started later for rural women. Possible reasons for such differences over time are discussed. PMID- 14526506 TI - Diagnosed mental and physical health conditions in the United States nursing home population: differences between urban and rural facilities. AB - CONTEXT: There has been limited examination of the differences in health characteristics of the rural long-term care population. Recognizing these differences will allow policymakers to improve access to long-term care services in rural communities. PURPOSE: To determine whether differences in likelihood of diagnosis exist between urban and rural nursing home residents for 8 common medical conditions: 4 mental health conditions (depression, anxiety, Alzheimer's, and non-Alzheimer's dementia) and 4 physical health conditions (cancer, emphysema/chronic obstructive pulmonary disease, heart disease, and stroke/transient ischemic attack). METHODS: We used multivariate logistic regression to examine data derived from the 1996 Nursing Home Component of the Medical Expenditure Panel Survey, a multistage stratified probability sample of 815 nursing homes and 5899 residents, representing 3.1 million individuals in the United States who spent 1 or more nights in nursing homes during 1996. FINDINGS: Residents in rural homes were less likely to be diagnosed with depression compared to those in homes in large metropolitan areas, and residents in homes in small metropolitan areas were less likely to have cancer than those in large metropolitan areas. Diagnostic status between urban and rural residents was comparable for the other 6 conditions. CONCLUSIONS: Further research is necessary to determine whether and why depression is inadequately diagnosed in rural nursing homes and to ascertain which types of cancer are responsible for the observed differential. Such research is particularly important for elderly nursing home residents who are more likely to suffer from chronic conditions that require significant medical supervision. PMID- 14526507 TI - The rural physician workforce in Florida: a survey of US- and foreign-born primary care physicians. AB - CONTEXT: To meet the challenge of primary care needs in rural areas, continuing assessment of the demographics, training, and future work plans of practicing primary care physicians is needed. PURPOSE: This study's goal was to assess key characteristics of primary care physicians practicing in rural, suburban, and urban communities in Florida. METHODS: Surveys were mailed to all of Florida's rural primary care physicians (n = 399) and a 10% sampling (n = 1236) of urban and suburban primary care physicians. FINDINGS: Responses from 1000 physicians (272 rural, 385 urban, 343 suburban) showed that rural physicians were more likely to have been raised in a rural area, foreign-born and trained, a National Health Service Corps member, or a J-1 visa waiver program participant. Rural physicians were more likely to have been exposed to rural medical practice or living in a rural environment during their medical school and residency training. Factors such as rural upbringing and medical school training did not predict future rural practice with foreign-born physicians. Overall, future plans for practice did not seem to differ between rural, urban, and suburban physicians. CONCLUSIONS: Recruiting and retaining doctors in rural areas can be best supported through a mission-driven selection of medical students with subsequent training in medical school and residency in rural health issues. National programs such as the National Health Service Corps and the J-1 visa waiver program also play important roles in rural physician selection and should be taken into account when planning for future rural health care needs. PMID- 14526508 TI - Emergency medical services in rural areas: the supporting role of state EMS agencies. AB - CONTEXT: EMS is an integral part of health care and is especially important in less densely populated areas. What is known about EMS in rural areas is limited because of fragmentation in the system and rudimentary data collection efforts. PURPOSE: The goal of this study is to identify important issues faced by rural EMS systems and describe the support of rural EMS providers by state EMS agencies. METHODS: A telephone survey of state EMS directors (response rate 95.7%) asked questions regarding issues in medical direction, programs, and initiatives by state EMS agencies that target rural and volunteer EMS providers, integration initiatives, and anticipated effects of the new Medicare fee schedule. FINDINGS: Medical direction in rural EMS was identified as a major issue for a majority of states. Integration in EMS is seen as a possible solution but does not occur very commonly. The survey found substantial variation in the state approach to EMS issues. Less than a third of the states in the study have a statewide EMS plan. State EMS agencies address rural EMS provider needs in a limited manner. EMS state agencies focus on regulation and funding of EMS providers, with only approximately a third providing technical assistance to EMS providers. CONCLUSIONS: The range in approaches to EMS issues at the state level will need to be taken into account in formulating national EMS policy. The limited provision of technical assistance leaves a void that may be addressed by other agencies and organizations in some states. In the absence of major federal funding initiatives, the development of EMS has become a state and local issue. A new national initiative may help address EMS issues and stimulate the development of EMS as a system beyond its current fragmented state. PMID- 14526509 TI - Rural and small-town attitudes about alcohol use during pregnancy: a community and provider sample. AB - CONTEXT: While there has been considerable research on prenatal alcohol use, there have been limited studies focused on women in rural and small-town environments. PURPOSE: This 2-part study examines gender differences in attitudes and perceived barriers to intervention in large community sample of persons living in rural and small-town environments in Kentucky (n = 3,346). The study also examines rural/small-town prenatal service providers' perceptions of barriers to assessment and intervention with pregnant substance abusers (n = 138). METHODS: Surveys were administered to a convenience sample of employees and customers from 16 rural and small-town community outlets. There were 1503 males (45%) and 1843 females (55%) ranging in age from under 18 years old to over 66 years old. Surveys also were mailed to prenatal providers in county health departments of the 13-county study area, with 138 of 149 responding. FINDINGS: Overall results of the community sample suggest that neither males nor females were knowledgeable about the harmful effects of alcohol use during pregnancy. Results also indicate substantial gender differences in alcohol attitudes, knowledge, and perceived barriers. Further, prenatal care providers identified several barriers in assessment and treatment of pregnant women with alcohol use problems in rural and small-town communities, including lack of knowledge and comfort with assessment as well as a lack of available and accessible treatment for referrals. PMID- 14526510 TI - Promoting pneumococcal immunizations among rural Medicare beneficiaries using multiple strategies. AB - CONTEXT: Vaccine-preventable diseases among adults are major contributing causes of morbidity and mortality in the United States. However, adult immunizations continue to be underutilized in both urban and rural areas. PURPOSE: To evaluate the effectiveness of a community-wide education campaign and mailed reminders promoting pneumococcal immunizations to rural Medicare beneficiaries. METHODS: We implemented a community-wide education campaign, and mailed reminders were sent to Medicare beneficiaries in 1 media market in Montana to increase pneumococcal immunizations. In a second distinct media market, mailed reminders only were sent to beneficiaries. FINDINGS: The proportion of respondents aged 65 years and older aware of pneumococcal immunizations increased significantly from baseline to follow-up among respondents both in the education-plus-reminder (63% to 78%, P = 0.04) and the reminder-only (64% to 74%, P = 0.05) markets. Overall from 1998 to 1999, there was a 3.7-percentage-point increase in pneumococcal immunization claims for Medicare beneficiaries in the education-plus-reminder market and a 1.5 percentage-point increase in the reminder-only market. Medicare beneficiaries sent reminders in the education-plus-reminder market compared to those in the reminder-only market were more likely to have a claim for pneumococcal immunization in 1999 (odds ratio 1.18, 95% confidence interval 1.08 to 1.28). The results suggest that these quality improvement strategies (community education plus reminders and reminders alone) modestly increased pneumococcal immunization awareness and pneumococcal immunization among rural adults. Mailed reminder exposure was associated with an increased prevalence of pneumococcal immunizations between 1998 and 1999 and was augmented somewhat by the education campaign. PMID- 14526512 TI - Southern Rural Access Program: an overview. AB - Rural residents experience significant disparities in health status and access to care. These disparities and access barriers are particularly prevalent in rural communities in the South. The Southern Rural Access Program, a national program of the Robert Wood Johnson Foundation, was designed as a long-term effort to improve access to basic health care in 8 of the most underserved states in the country. The program was launched in 1998 with 3 goals: (1) to increase the supply of providers in underserved areas, (2) to strengthen the health care infrastructure, and (3) to build capacity at the state and community level to solve problems. The first 3-year phase of the program made $13.8 million available to communities in the 8 target states, and a January 2002 reauthorization of the program will make an additional $18.9 million available in the next 4 years. This article will provide an overview of the Southern Rural Access Program, focusing on the development and evolution of the program during its first 3-year phase. The article will also highlight some of the refinements that the foundation has made during the 2002-2006 second phase of the program. PMID- 14526511 TI - Children's health insurance status, access to and utilization of health services, and unmet health needs in a rural Alabama school system. AB - PURPOSE: This study examines the relationship between children's health insurance status and utilization of health services, establishment of a medical home, and unmet health needs over a 3-year period (1996-1998) in a rural Alabama K-12 school system. METHODS: As part of a children's health insurance outreach program, questionnaires were administered to parents of 754 children regarding health and health care access. In addition, noninvasive head-to-toe physical assessments of children were conducted on-site at 4 schools. FINDINGS: A relationship between health care utilization and insurance status was observed. Results found that insured children had 1.183 (P < .0115) times the number of medical visits as uninsured children. Among uninsured children, the time since last dental visit was 1.6 (P < .001) times longer than that of insured children. Also, insured children were 5.21 times more likely than uninsured (P < .0001) to report having a medical home. No significant differences between insured and uninsured children were found regarding unmet health needs as measured by referrals made after the children's physical assessments. CONCLUSIONS: Child health coverage is an important determining factor in the ability of families to access and utilize health care services. These findings have implications for populations in similar rural communities across the nation. PMID- 14526513 TI - Use of program logic models in the Southern Rural Access Program evaluation. AB - The Southern Rural Access Program (SRAP) evaluation team used program logic models to clarify grantees' activities, objectives, and timelines. This information was used to benchmark data from grantees' progress reports to assess the program's successes. This article presents a brief background on the use of program logic models--essentially charts or diagrams specifying a program's planned activities, objectives, and goals--for evaluating and managing a program. It discusses the structure of the logic models chosen for the SRAP and how the model concept was introduced to the grantees to promote acceptance and use of the models. The article describes how the models helped clarify the program's objectives and helped lead agencies plan and manage the many program initiatives and subcontractors in their states. Models also provided a framework for grantees to report their progress to the National Program Office and evaluators and promoted the evaluators' visibility and acceptance by the grantees. Program logics, however, increased grantees' reporting requirements and demanded substantial time of the evaluators. Program logic models, on balance, proved their merit in the SRAP through their contributions to its management and evaluation and by providing a better understanding of the program's initiatives, successes, and potential impact. PMID- 14526514 TI - A description of the Southern Rural Access Program's practice management strategies. AB - CONTEXT: Many state, federal, and foundation resources have been invested in improving the recruitment of primary care providers to rural communities. The Southern Rural Access Program of the Robert Wood Johnson Foundation (RWJF) has provided varying levels of support to several southern states to assist with retention of those providers. PURPOSE: This study describes the strategies that 6 states used to develop and implement practice management technical assistance services for rural health care providers. METHODS: Practice managers in each of the 6 states were surveyed regarding how their service was structured, what types of entities were eligible, and the nature of the technical assistance offered. Information regarding what types of entities used the service, characteristics of the practices, and the number of practices served was also collected. FINDINGS: The survey results showed that almost half (46%) of all practices assisted were private stand-alone physician practices, with overall practice assessments being the practice management service rendered most often. Although the type of organisational home for the technical assistance services varied by state, overall states employed an average of 1.67 full-time equivalent practice managers (0.81 full-time equivalent supported by RWJF) and received an average of $136,055 per state from the RWJF for the 2-year period beginning April 2002 for practice management support. CONCLUSIONS: Overall, the study found that the type of organizational home did not appear to affect the type of technical assistance services offered. However, the type of orgnizational home did appear to affect what types of providers used the service, with trade associations assisting their members or constituents at least half the time. PMID- 14526515 TI - Rural and urban physicians' perceptions regarding the role and practice of the nurse practitioner, physician assistant, and certified nurse midwife. AB - CONTEXT: There is a dearth of literature citing the differences in rural and urban physicians' perceptions of the role and practice of nurse practitioners, physician assistants, and certified nurse midwives (nonphysician providers). PURPOSE: The purpose of this study was to investigate and compare differences, if any, between rural and urban primary care physicians' perceptions of the role and practice of nonphysician providers. RESULTS: Despite a 15.55% response rate using a mail-out survey in South Carolina, data from 681 rural and urban primary care physicians indicated that they perceived that nonphysician providers possess the necessary skills and knowledge to provide primary care to patients, are an asset to a physician's practice, free the physician's time to handle more critically ill patients, and increase revenue for the practice, but increase the risk of patient care mistakes and a physician's time in administrative duties. Urban physicians' mean scores were higher for perceiving that nonphysician providers are able to see as many patients in a given day as a physician but experience impediments in the delivery of patient care. CONCLUSIONS: Results will be used to clarify physicians' perceptions regarding the role and practice of nonphysician providers to reduce impediments to patient care access. PMID- 14526516 TI - A comparative assessment of West Virginia's financial incentive programs for rural physicians. AB - CONTEXT: Financial incentive programs are increasingly being used as a strategy to recruit physicians to underserved rural areas. Critical evaluation of state supported programs is often lacking but is necessary to determine their efficacy and to improve outcomes. PURPOSE: The purpose of this study was to assess 4 service-contingent programs in West Virginia, a state with critical physician shortages. METHODS: Survey instruments were developed to evaluate the effectiveness of these programs and to document the practice environments and career paths of obligated allopathic and osteopathic physicians compared with a control group of nonobligated rural practitioners. Data were also collected from physicians who were recipients of multiple incentive programs and from obligated physicians who had defaulted. FINDINGS: Responses from more than 60% of surveyed physicians indicated that the typical respondent was a married white male who was a midcareer family practice physician. Obligated physicians were more likely than nonobligated physicians to have graduated from a West Virginia medical school and residency program, to be influenced by financial factors in their career decisions, to provide care to uninsured patients, and to work in offices that offered sliding fee scales. Both groups of physicians demonstrated similar retention patterns, reported a high degree of job satisfaction, and expressed a need for more practice management training. CONCLUSIONS: Although these financial incentive programs were found to be effective in recruiting primary care physicians to medically underserved areas of the state, the financial support of these programs was found to be too modest, and improved marketing of the programs was indicated. PMID- 14526517 TI - Recruitment of rural health care providers: a regional recruiter strategy. AB - CONTEXT: Access to care in rural areas is a major problem. Despite more than 20% of the US population residing in these areas, only 9% of physicians practice there. Extensive research has documented multiple issues that affect where physicians decide to locate and maintain practices. Creative strategies have been used to influence these recruitment and retention decisions. An emerging strategy, borne out of the Robert Wood Johnson Foundation's Southern Rural Access Program (SRAP), effectively uses a targeted regional approach to assist rural communities and health care facilities in assessing health care needs and recruiting primary care providers. PURPOSE: This article examines the issues surrounding recruitment and retention of primary care providers to rural areas and describes the experiences of the regional recruitment strategy in several states and in particular in the Mississippi Delta region of Arkansas. METHODS: A case study approach is used to examine the targeted regional recruiter strategy in the Mississippi Delta region of Arkansas. FINDINGS: The regional recruiter strategy, which combines traditional recruitment efforts with community development activities, has been successful in recruiting health care providers to rural communities. The cost-effective strategy can be easily replicated in other rural states. CONCLUSIONS: Community factors affect provider decisions on practice locations. Addressing community factors in recruitment efforts through community development activities may increase their success. PMID- 14526518 TI - A community development approach to rural recruitment. AB - Programs designed to empower rural communities for health care provider recruitment have usually focused on the health care sector without aggressively addressing broader community development issues. The Recruitable Community Project (RCP) in West Virginia includes community education on recruiting and also assessments of and recommendations to rural communities on broad-based community development, aiming to enhance communities' recruiting potential. The project provides multidisciplinary university-based planning assistance programs for small communities, involving collaborative community visits. The project also uses a project manager as a "community encourager" who participates in community education and in the formulation of sustained community recruiting efforts. From August 1999 through August 2001, 7 underserved rural communities completed the RCP organizational processes and hosted planning assistance teams. Members of community recruitment boards gave high marks to the RCP process, its planning assistance teams, and its usefulness in establishing community ties to state and academic agencies. Since working with the RCP, the 7 communities have recruited 27 providers, success possibly stimulated by their RCP involvement (data current as of September 2002). This model of community training and development to empower rural communities to better recruit health professionals shows early promise. This model could be broadened to include more collaboration of community development and health science disciplines programs for recruitment and retention efforts. PMID- 14526519 TI - The Southern Rural Access Program and Alabama's Rural Health Leaders Pipeline: a partnership to develop needed minority health care professionals. AB - Rural Health Leaders Pipeline programs are intended to increase the number of youth interested in and pursuing health professions in rural communities. This paper presents 2 complementary approaches to Rural Health Leaders Pipeline programs. Two different organizations in Alabama recruit students from 18 specified counties. One organization is a rural, community-based program with college freshmen and upperclassmen from rural communities. Students shadow health professionals for 6 weeks, attend classes, visit medical schools, complete and present health projects, and receive support from online tutors. The second organization is a university-based program that supplements an existing 11th grade-medical school rural medicine pipeline with 10 minority students from rural communities who have graduated from high school and plan to enter college as premedical students in the following academic year. Students participate in classes, tutorials, seminars, and other activities. Students earn college credits during the 7-week program, maintain contact with program staff during the school year, and by performance and interest can continue in this pipeline program for a total of 4 consecutive summers, culminating in application to medical school. Each organization provides stipends for students. Early experiences have been positive, although Rural Health Leaders Pipeline programs are expensive and require long-term commitments. PMID- 14526521 TI - Connecting our resources: Louisiana's approach to community health network development. AB - Louisiana's rural community health systems are in crisis because of pressures fueled by the rising costs of health care, sustained poor health status, state budget shortfalls and changes in priorities, and a sliding rural economy. The development of community health networks is providing new infrastructure and capacity for communities to reprioritize, formulate innovative partnerships, and leverage new resources. Successful elements of Louisiana's network development experience include community commitment to engage in study and action; the availability of capable and motivated technical assistance; an approach that involves open-engagement, community-driven decision-making; and data-driven problem definition, prioritization, and solutions. Louisiana's experiences illustrate the benefits of developing networks along with, or as a result of, a community health plan. When a community owns its health improvement plan, it is more likely to support the new network as a structure for implementation. Broad scale participation is also a principle of success. When social service agencies are included along with health agencies, more comprehensive strategies result, and they bring additional resources, resulting in more holistic solutions. The cases of 2 networks are presented as illustrations. One involves the facilitation of a community planning process for an existing network. The plan helped to expand the network's community connections and support and provided the content for a successful application for a Health Resources and Services Administration Community Access Program grant. In the second case, a new network was developed, and it leveraged federal funds from the federal Office of Rural Health Policy's Network Development Grant Program. PMID- 14526520 TI - Transforming the delivery of rural health care in Georgia: state partnership strategy for developing rural health networks. AB - Since 1996, 19 networks covering 74 of the 117 rural counties in Georgia have emerged. This grassroots transformation of rural health care occurred through a series of partnerships launched by state government officials. These partnerships brought together national and state organizations to pool resources for investment in an evolving long-term strategy to develop rural health care networks. The strategy leveraged resources from partners, resulting in greater impact. Change was triggered and accelerated using an intensive, flexible technical assistance effort amplified by developmental grants to communities. These grants were made available for structural and organizational change in the community that would eventually lead to improved access and health status. Georgia's strategy for developing rural health networks consisted of 3 elements: a clear state vision and mission; investment partnerships; and proactive, flexible technical assistance. Retrospectively, it seems that the transformation occurred as a result of 5 phases of investment by state government and its partners. The first 2 phases involved data gathering as well as the provision of technical assistance to individual communities. The next 3 phases moved network development to a larger scale by working with multiple counties to create regional networks. The 5 phases represent increasing knowledge about and commitment to the vision of access to care and improved health status for rural populations. PMID- 14526522 TI - The Arkansas River Valley Rural Health Cooperative: building a three-pronged approach to improved health and health care. AB - This paper describes the Arkansas River Valley Rural Health Cooperative (ARVRHC), one of the Arkansas networks jump-started with support from the Southern Rural Access Program (SRAP). The initial goal of the network was to develop a subsidized health insurance program to provide affordable medical services for the uninsured population (23%) in the 3-county service area. When planning efforts called for the network to address broader needs, the ARVRHC crafted a more comprehensive 3-pronged program model consisting of 3 interrelated programs: (1) the Health Care Access Program (HCAP), (2) the Health Education and Disease Management Program (HE&DMP), and 3) the Information and Assistance Program (I&AP). The HCAP is designed to address the financial barriers to access through a community-based health plan. The HE&DMP focuses on improving the health of individuals through education, counseling, and preventive care. The I&AP links low-income families to existing public assistance programs (e.g., Medicaid) and social support services. The Prescription Drug Assistance Program is one of the I&AP programs that helps individuals without prescription coverage obtain drugs at no cost. A key lesson learned is the importance of combining technical assistance with funding. The ARVRHC has been successful in leveraging funding, having received over $1.7 million in grant funds since 1999. A critical challenge facing the network today is the need for ongoing subsidy funding. Proposed legislation for a federal demonstration of the HCAP and similar programs would enable full implementation and evaluation of this model. PMID- 14526523 TI - Improving access to capital for health care infrastructure: the experience of the Southern Rural Access Program's revolving loan fund. AB - Lack of access to affordable capital is a formidable barrier that compromises rural health care infrastructure development in poor rural areas. Commercial lending institutions are often limited in their ability to respond to those needs due to traditional lending criteria: creditworthiness, equity, management ability, experiences, and cash flow or profits. In the Southern Rural Access Program, a development model more frequently used in other sectors has been successfully applied to health care to help clear these hurdles. This paper describes the 5 operational loan funds in Arkansas, Louisiana, Mississippi, South Carolina, and West Virginia receiving support from the Southern Rural Access Program. Two models of loan funds have evolved: those led by health agencies and those led by community development finance institutions whose mission is rural economic development. This paper outlines major distinctive features of these 2 approaches and describes major implementation challenges these loan funds face. Key accomplishments are high-lighted, including the ability to leverage additional resources from state, federal, philanthropic, and private sources through these funds. These loan fund programs provide models for other states interested in improving access to capital to help build the rural health care infrastructure while making health care more economically viable through integration with other community development initiatives. PMID- 14526524 TI - Getting from here to there: evaluating West Virginia's rural nonemergency medical transportation program. AB - With funding from the 21st Century Challenge Fund, the West Virginia Rural Health Access Program created Transportation for Health, a demonstration project for rural nonemergency medical transportation. The project was implemented in 3 sites around the state, building on existing transportation systems--specifically, a multicounty transit authority, a joint senior center/transit system, and a senior services center. An evaluation of the project was undertaken to answer 3 major questions: (1) Did the project reach the population of people who need transportation assistance? (2) Are users of the transportation project satisfied with the service? (3) Is the program sustainable? Preliminary results from survey data indicate that the answers to questions 1 and 2 are affirmative. A break-even analysis of all 3 sites begins to identify programmatic and policy issues that challenge the likelihood of financial sustainability, including salary expenses, unreimbursed mileage, and reliance on Medicaid reimbursement. PMID- 14526525 TI - Smile Alabama! Initiative: interim results from a program to increase children's access to dental care. AB - Alabama faced an oral health crisis, with decreasing dental provider participation and increasing enrollment of Medicaid-eligible children. In response, the Smile Alabama! initiative was designed to improve oral health care services for Medicaid-eligible children by increasing the number of participating dentists by 15% and the number of children receiving dental care annually by 5% by January 31, 2004. The initiative is composed of 4 specific components: claims processing, dental reimbursement, provider education and recruitment, and recipient education. Specific interventions were implemented for each component. From fiscal year 1999 to fiscal year 2002, enrollment of targeted Medicaid children increased 32.7%. During this same period, the number of participating dental providers in the Alabama Medicaid dental program increased by 127 providers, a 38.7% increase. The number of children receiving dental services increased from 82,600 in fiscal year 1999 to 130,208 in fiscal year 2002, a 57.1% total increase, with a 4.8% increase in the annual dental visit rate. The experience suggests that access to oral health care services can be improved through a multidimensional, strategically planned dental outreach initiative in spite of dramatic increases in Medicaid enrollment. PMID- 14526526 TI - Lessons learned in phase I of the Southern Rural Access Program. AB - The Robert Wood Johnson Foundation's Southern Rural Access Program has been an important investment of philanthropic funds to augment resources and improve health care access in underserved rural communities. The program's first phase has taught important lessons about building capacity in rural health care. This article uses a variety of data to document the program's major accomplishments and most significant challenges to date. The program's revolving loan fund efforts are promising. The program has also played a catalytic role in stimulating rural health network development in the South and has helped stimulate partnerships with Southern philanthropies and multiple local, state, and federal agencies. Challenges have included the broad geographic and programmatic focus of the initiative as well as changing and often difficult state policy environments. Additional challenges include maintaining interagency coordination over time and managing staff and lead agency turnover. Overall, the experience suggests that a concentrated regional approach has merit. PMID- 14526527 TI - Physician practice groups--the effect of Clackamas. PMID- 14526529 TI - DoctorQuality analyzes impact of reducing restraints in medical organizations. AB - In recent years, many hospitals and some state agencies with healthcare oversight have called for the creation of restraint-free environments. But can these changes lead to an actual increase in falls? A new web-based system that uses real-time data is providing a picture of the impact of reduced restraint use. PMID- 14526528 TI - Electronic health records: turning research into decision support for quality care. AB - Much has been written and discussed about low clinical and decision-support tools can help providers deliver quality care to their patients. But there are theories -and then there are realities--about what will actually work in the real world and how useful they will be. When incorporating decision-support tools into electronic health records, the results may not always be predictable. PMID- 14526530 TI - New physician recognition program for cardiovascular care announced. AB - Physicians and medical groups that provide quality care to patients with heart disease or stroke now can be recognized under a program launched last month by the National Committee for Quality Assurance and the American Heart Association/American Stroke Association. PMID- 14526531 TI - [Copy number of ribosomal operons in prokaryotes and its effect on phylogenic analyses]. AB - Different aspects of the presence of multiple copies of ribosomal operons in prokaryotic genomes are reviewed. Structure of prokaryotic ribosomal operons is briefly described. The available data are summarized regarding the copy number of ribosomal genes in various prokaryotic genomes, the degree of polymorphism of their individual copies, physiological and evolutionary aspects of the presence of the multiple copies of ribosomal genes. The review also considers the influence of the presence of multiple copies of ribosomal genes on the results of identification of prokaryotic isolates and of the studies of prokaryotic diversity in environmental samples based on phylogenetic analysis of 16S rRNA gene sequences. PMID- 14526532 TI - [The alternative oxidase of Yarrowia lipolytica mitochondria is unable to compete with the cytochrome pathway for electrons]. AB - The activity of the cyanide-resistant alternative oxidase (pathway) of Y. lipolytica mitochondria was studied as a function of the activity of the major, cyanide-sensitive, cytochrome pathway. The contribution of the alternative oxidase to the total respiration of mitochondria was evaluated by measuring the rate of oxygen consumption in the presence of cyanide (an inhibitor of the cytochrome pathway). The potential activity of the cytochrome pathway was evaluated spectrophotometrically, by measuring the oxidation rate of cytochrome c by ferricyanide, which accepts electrons from complex III (cytochrome c) of this pathway. The oxidation of succinate by mitochondria in the presence of ferricyanide and cyanide was accompanied by oxygen consumption due to the transfer of electrons through the alternative pathway. The subsequent addition of ADP or FCCP (an uncoupler of oxidative phosphorylation in the cytochrome pathway) completely inhibited the consumption of oxygen by the mitochondria. Under these conditions, the inhibition of the alternative pathway by benzohydroxamic acid failed to affect the transfer of electrons from cytochrome c to ferricyanide. Benzohydroxamic acid did not influence the rate of ferricyanide reduction by the cytochrome pathway occurring in controlled state 4, nor could it change the phosphorylation quotient ATP/O upon the oxidation of various substrates. These findings indicate that the alternative pathway is unable to compete with the cytochrome respiratory chain for electrons. The alternative pathway transfers only electrons that are superfluous for the cytochrome chain. PMID- 14526533 TI - [Central metabolism in Acinetobacter sp. grown on ethanol]. AB - The ethanol-grown cells of the mutant Acinetobacter sp. strain 1NG, incapable of producing exopolysaccharides, were analyzed for the activity of enzymes of the tricarboxylic acid (TCA) cycle and some biosynthetic pathways. In spite of the presence of both key enzymes (isocitrate lyase and malate synthase) of the glyoxylate cycle, these cells also contained all enzymes of the TCA cycle, which presumably serves biosynthetic functions. This was evident from the high activity of isocitrate dehydrogenase and glutamate dehydrogenase and the low activity of 2 oxoglutarate dehydrogenase. Pyruvate was formed in the reaction catalyzed by oxaloacetate decarboxylase, whereas phosphoenolpyruvate (PEP) was synthesized by the two key enzymes (PEP carboxykinase and PEP synthase) of gluconeogenesis. The proportion between these enzymes was different in the exponential and the stationary growth phases. The addition of the C4-dicarboxylic acid fumarate to the ethanol-containing growth medium led to a 1.5- to 2-fold increase in the activity of enzymes of the glyoxylate cycle, as well as of fumarate hydratase, malate dehydrogenase, PEP synthase, and PEP carboxykinase (the activity of the latter enzyme increased by more than 7.5 times). The data obtained can be used to improve the biotechnology of production of the microbial exopolysaccharide ethapolan on C2-substrates. PMID- 14526534 TI - [The activity of xylose reductase and xylitol dehydrogenase in yeasts]. AB - The activity and the cofactor specificity of xylose reductase and xylitol dehydrogenase were studied in extracts of yeasts from the genera Candida, Kluyveromyces, Pachysolen, Pichia, and Torulopsis grown under microaerobic conditions. It was found that xylitol dehydrogenase in all of the yeast species studied is specific for NAD+; xylose reductase in the xylitol-producing species C. didensiae, C. intermediae, C. parapsilosis, C. silvanorum, C. tropicalis, Kl. fragilis, Kl. marxianus, P. guillermondii, and T. molishiama is specific for NADPH; and xylose reductase in the ethanol-producing species P. stipitis, C. shehatae, and Pa. tannophilus is specific for both NADPH and NADH. PMID- 14526536 TI - [The absence of direct relationship between the ability of yeasts to grow at elevated temperatures and their survival after lethal heat shock]. AB - The study of the growth of the yeasts Rhodotorula rubra, Saccharomyces cerevisiae, and Debaryomyces vanriji at elevated temperatures and their survival after transient lethal heat shock showed that the ability of these yeasts to grow at supraoptimal temperatures (i.e., their thermoresistance) and their ability to tolerate lethal heat shocks (i.e., their thermotolerance) are determined by different mechanisms. The thermotolerance of the yeasts is suggested to be mainly determined by the division rate of cells before their exposure to heat shock. PMID- 14526535 TI - [Biochemical characterization of the yeast Yarrowia lipolytica overproducing carboxylic acids from ethanol. Nitrogen metabolism enzymes]. AB - A comparative assay of nitrogen metabolism enzymes in the Yarrowia lipolytica mutant N1 grown under conditions promoting the overproduction of either alpha ketoglutaric acid (KGA) or citric acid showed that the overproduction of KGA correlates with an increase in the activities of the NAD- and NADP-linked glutamate dehydrogenase, glutamic-pyruvic transaminase, and glutamic-oxaloacetic transaminase reactions. These reactions are likely to be responsible for the overproduction of KGA by this mutant. In contrast, the overproduction of citric acid correlated with a decline in the activities of the NAD- and NADP-linked glutamate dehydrogenases and with an increase in the activities of glutamine synthetase and glutamate synthase. PMID- 14526537 TI - [The extracellular protein of Luteococcus japonicus subsp casei reactivates cells inactivated by ultraviolet radiation or heating]. AB - The culture liquid of Luteococcus japonicus subsp. casei was found to be able to reactivate cells of this bacterium inactivated by UV irradiation or heat shock. The antistress activity of the culture liquid was due to the presence of an extracellular exometabolite of a protein nature with a molecular mass of more than 10 kDa. When the bacterium was grown in nutrient broth or glucose-containing mineral medium, the antistress protein was secreted by cells in the logarithmic growth phase. The reactivating effect of the antistress protein was inversely proportional to the survival rate of stressed cells. PMID- 14526538 TI - [Photodynamic damage of yeast subcellular fraction induced by elevated levels of endogenous protoporphyrin IX]. AB - The 2,2'-dipyridyl-induced accumulation of protoporphyrin IX in Saccharomyces cerevisiae cells was shown to be accompanied by the photoinhibition of cell respiration and the enhancement of the photoinduced permeability of plasma membranes to the fluorescent dye primuline. The visible-light illumination (at 400-600 nm) of the mitochondria and plasma membranes isolated from yeast cells with a high level of endogenous protoporphyrin IX intensified lipid peroxidation in these subcellular organelles. Comparative studies showed that the rad 52 mutant cells, which are deficient in the postreplicative recombinational DNA repair system, are considerably more sensitive to the inactivating action of visible light than are the wild-type cells and the rad 3 mutant cells, which are deficient in the excision DNA repair system. The contribution of photodynamic damage to the yeast subcellular organelles to the lethal photodynamic effect is discussed. PMID- 14526539 TI - [Leptospirillum-like bacteria and their role in pyrite oxidation]. AB - Two strains of Leptospirillum-like bacteria isolated from dumps of Alaverdi and Akhtala sulfide ore deposits in Armenia were studied. The optimum and maximum temperatures for the growth of both strains were 37 and 40 degrees C, respectively. The pH optimum was 2.0-2.3. Bacterial growth and ferrous iron oxidation were inhibited by yeast extract. The pyrite-leaching activity of the Leptospirillum-like bacteria under mesophilic conditions was close to that of Acidithiobacillus ferroxidans and exceeded by 2.0-2.7 times the activity of these moderately thermophilic bacteria at 37 degrees C. The leaching of pyrite by Leptospirillum-like bacteria increased in the presence of sulfur-oxidizing bacteria, particularly, in their association with a thermotolerant sulfur oxidizing bacterium. PMID- 14526540 TI - [The extracellular proteases of the phytopathogenic bacterium Xanthomonas campestris]. AB - The culture liquids of three Xanthomonas campestris pv. campestris strains were found to possess proteolytic activity. The culture liquid of strain B-611 with the highest proteolytic activity was fractionated by salting-out with ammonium sulfate, gel filtration, and ion-exchange chromatography. The electrophoretic analysis of active fractions showed the presence of two proteases in the culture liquid of strain B-611, the major of which being serine protease. The treatment of cabbage seedlings with the proteases augmented the activity of peroxidase in the cabbage roots by 28%. PMID- 14526541 TI - [Zygote formation in Blakeslea trispora: morphological peculiarities and relationship with carotenoid synthesis]. AB - Changes associated with zygospore formation in the mucorous fungus Blakeslea trispora were studied. Zygospores are dormant cells with thickened cell walls and large central lipid vacuoles containing large amounts of lycopene. We established for the first time that B. trispora gametangia of different sexes differ in their carotenoid content and revealed that zygote formation involves a novel structure that consists of densely intertwined hyphae. Using inhibitory analysis (blocking beta-carotene synthesis with diphenylamine and 2-amino-6-methylpyridine), we showed that suppression of carotene production results in the inhibition of zygote formation. Hence, we established a manifest dependence of zygote formation on beta-carotene synthesis. PMID- 14526542 TI - [The cell wall teichoic acids of streptomycetes from the "Streptomyces cyaneus" cluster]. AB - The cell wall anionic polymers of the 13 species of the "Streptomyces cyaneus" cluster have a similar structure and contain beta-glucosylated 1,5-poly(ribitol phosphate) and 1,3-poly(glycerol phosphate). In the degree of glucosylation of the ribitol phosphate units of their teichoic acids, the cluster members can be divided into two groups. The streptomycetes of the first group (S. afghaniensis, S. janthinus, S. purpurascens, S. roseoviolaceus, and S. violatus) are characterized by a very similar structure of their cell walls, completely glucosylated 1,5-poly(ribitol phosphate) chains, and a high degree of DNA homology (67-88%). The cell wall teichoic acids of the second group (S. azureus, S. bellus, S. caelestis, S. coeruleorubidus, S. curacoi, and S. violarus) differ in the degree of beta-glucosylation of their 1,5-poly(ribitol phosphate) chains and have a lower level of DNA homology (54-76%). Two streptomycetes of the cluster (S. cyaneus and S. hawaiiensis) are genetically distant from the other cluster members but have the same composition and structure of the cell wall teichoic acids as the second-group streptomycetes. The data obtained confirm the genetic relatedness of the "S. cyaneus" cluster members and suggest that the structure of the cell wall teichoic acids may serve as one of the taxonomic criteria of the species-level status of streptomycetes. PMID- 14526543 TI - [Changes in the lipid composition of Mucor hiemalis sporangiospores related to the age of the spore-forming culture]. AB - Analysis of sporangiospore lipids of the fungus Mucor hiemalis F-1156 showed that alterations occur in the content of fatty acids and individual classes of lipids during long-term cultivation (for about 20 days). The changes in the lipid composition related to the age of the spore-forming mycelium suggest an important role of sporangiospore lipids in spore germination and in further development of the spherical cells formed in this processes. The M. hiemalis F-1156 sporangiospores with a lipid pool exhausted during long-term cultivation can give rise to both mycelial and yeastlike growth. PMID- 14526544 TI - [Effect of culture conditions on growth and adhesion of Bacillus licheniformis]. AB - Adhesion to glass of actively growing cells of the thermophilic Bacillus licheniformis, isolated from the Medyaginskaya test borehole (Yaroslavl' oblast), was studied. The reversible adhesion (RA) manifests itself in a decline of cell density (short of cell lysis) in the liquid culture over the first 20-40 min of growth followed by normal exponential growth. The RA is minimal under favorable growth conditions but increases when cells are transferred to a new medium, especially one with a pH, temperature, salinity, or concentration of Ca2+ ions nonoptimal for the given species. Under unfavorable growth conditions, the adhesion becomes irreversible. The obtained data suggest that RA represents an adaptation mechanism important for population survival. PMID- 14526545 TI - [Comparative study of microbial communities from cultured and natural population of the mussel Mytilus trossulus in Peter the Great bay]. AB - The 525 strains of heterotrophic bacteria isolated from natural and cultured populations of the mussel Mytilus trossulus and the surrounding seawater were identified to a genus level on the basis of phenotypic analysis and the fatty acid composition of cell lipids. Gram-negative isolates were dominated by six genera of the family Enterobacteriaceae and by the genera Pseudoalteromonas, Vibrio, Photobacterium, Cytophaga/Flavobacterium/Bacteroides, Pseudomonas, and Moraxella, Gram-positive isolates were mainly represented by the genus Streptomyces. The taxonomic compositions of natural and cultured populations of the mussel M. trossulus in Peter the Great Bay were similar. PMID- 14526546 TI - [Microbial sulfate reduction in sediments of the coastal zone and littoral of the Kandalaksha bay of the White sea]. AB - Microbiological and biogeochemical investigations of the coastal zone and the littoral of the Kandalaksha Bay of the White Sea were carried out. The material for investigations was obtained in the series of expeditions of the Institute of Microbiology, Russian Academy of Sciences, in August 1999, 2000, 2001, and in March 2003. The studies were conducted on the littoral and in the water area of the Kandalaksha Preserve, the Moscow University Belomorsk Biological Station, and the Zoological Institute Biological Station, Russian Academy of Sciences, Sediment sampling on the littoral was carried out in the typical microlandscapes differing in the sediment properties and macrobenthos distribution. The maximal sulfate reduction rate (SRR) was shown for the shallow part of the Chemorechenskaya Bay (up to 2550 micrograms S/(dm3 day)) and in the Bab'ye More Bay (up to 3191 micrograms S/(dm3 day)). During the winter season, at a temperature of -0.5-0.5 degrees C, the SRR in the sediments of the Kartesh Bay was 7.9-13 micrograms S/(dm3 day). In the widest limits, the SRR values varied in the sediment cores sampled on the littoral. The minimal values (11 mu]g S/(dm3 day)) were obtained in the core samples on the silt-sandy littoral. The littoral finely dispersed sediments rich in organic matter were characterized by high SRR values (524-1413 micrograms S/(dm3 day)). The maximal SRR values were shown for the sediments present within the stretch of decomposing macrophytes, in local pits at the lower littoral waterline, and in the mouth of a freshwater stream (51 159 mg S/(dm3 day)). A sharp difference in the level of H2S production in the type microlandscapes was shown. The average hydrogen sulfide production in finely dispersed sediments constituted 125 mg S/(m2 day); in stormy discharge deposits, 1950 mg S/(m2 day); in depressions under stones and in silted pits, 4300 mg S/(m2 day). A calculation made with regard to the area of microlandscapes with increased productivity shows that the daily H2S production per 1 km2 of the littoral (August) is 60.8 to 202 kg S/(km2 day), while the organic carbon consumption for sulfate reduction per 1 km2 of the littoral is 46 to 152 kg C(org)/(km2 day). PMID- 14526548 TI - [The quantity and structure of the root-associated microbial complexes of two greenhouse rose cultivars]. AB - The study of the root-associated microbial complexes of affected and healthy rose plants of two cultivars (Grand gala and Royal velvet) grown in a greenhouse showed that the biomass of eukaryotic microorganisms in the rhizoplane and rhizosphere of healthy rose plants and in the surrounding soil was considerably lower than in the same loci of affected plants. In contrast, the biomass of root associated prokaryotic microorganisms was higher in the case of healthy than in the case of affected rose plants. The root-associated bacterial complexes of both affected and healthy rose plants were dominated by the genera Arthrobacter, Rhodococcus, and Myxobacterium and did not contain phytopathogenic bacteria. The root-associated fungal complex of healthy roses was dominated by fungi of the genus Trichoderma, whereas that of the affected rose plants was dominated by the species Aureobasidium microstictum. The affected cane cuttings and cankers occurring on affected canes were found to contain Coniothyrium fuckelii (the causal fungus of rose stem canker) and sclerotia of Botrytis cinerea (the causal fungus of gray rot). The micromycete complex of healthy rose plants was not so diverse as was the micromycete complex of affected rose plants. PMID- 14526547 TI - [Methanogenic sarcina from an anaerobic microbial community degrading p-toluene sulfonate]. AB - The methanogenic strain MM isolated from an anaerobic microbial community degrading p-toluene sulfonate showed optimal values of temperature and pH for growth equal to 37 degrees C and 6.3-6.9, respectively. The doubling times of the isolate grown on methanol, acetate, and methylamines under the optimal conditions were 8.8, 19.1, and 10.3-28.1 h, respectively. The growth of strain MM was observed only when the cultivation medium contained casamino acids or p-toluene sulfonate. The G + C content of the DNA of the isolate was 40.3 mol%. This, together with DNA-DNA hybridization data, allowed the new isolate to be identified as a strain of the species Methanosarcina mazei. The new isolate differed from the known representatives of this species in that it was resistant to alkylbenzene sulfonates and able to demethylate p-toluene sulfonate when grown on acetate. PMID- 14526549 TI - [Variety-specific actinomycete complexes associated with barley roots in soddy podzolic soil]. AB - The root-colonizing actinomycete complexes of genotypically different barley plants grown in soddy podzolic soil were found to contain streptomycetes of the sections Cinereus (series Chromogenes, Achromogenes, and Aureus) and Roseus (series Fradiae), dominant being streptomycetes of the section Cinereus ser. Chromogenes. The abundance and diversity of soil streptomycetes in the barley rhizoplane increased in the order: var. 999-93 < var. Kumir < var. Novichok < var. 889-93. Experiments revealed functional specificity in the root-associated actinomycete complexes of different barley varieties. The actinomycete complex colonizing the barley var. 999-93 roots was distinguished by a wide range of utilizable root exudate metabolites and a low occurrence rate of antagonistic species. PMID- 14526550 TI - [Morphology and dynamics of biofilms formed by Bacillus subtilis transgenic strain]. PMID- 14526551 TI - Ethics and medical information. PMID- 14526552 TI - Benign positional paroxysmal vertigo: videonystagmographic study using rotatory test. AB - Benign Peripheral Paroxysmal Vertigo is a disease of the posterior labyrinth caused by endolymphatic debris, provoking vertigo with some movements of the head. Diagnosis is usually made by finding the positional nystagmus with appropriate manoeuvres. Spontaneous resolution is frequent and in these cases diagnosis is only probable and suspected from anamnesis. Aim of the present investigation was to establish more evaluation parameters in the study of Benign Peripheral Paroxysmal Vertigo. A series of 97 selected patients presenting Benign Peripheral Paroxysmal Vertigo, have been submitted to sinusoidal kinetic test. Patients have been studied during the acute phase of the condition and after recovery. Vestibulo-oculomotor reflex has been sought by stimulating the horizontal and vertical canals. Kinetic stimulus consisted in sinusoidal rotation at 0.12 Hz and 0.05 Hz. Evaluation parameters comprised preponderance, gain and phase of provoked nystagmus, recorded by means of an Ulmer videonystagmograph. Using this same technique of stimulation, 20 normal volunteers were studied in order to establish normal values for reference. Values obtained in the patient population of patients have been compared, by Student t test, with values obtained in the same cured patients and with those in normal subjects. In the patients with Benign Peripheral Paroxysmal Vertigo of the lateral canal a nystagmus preponderance toward the healthy side was observed, as well as an increase in the phase lead, also in the canals not affected by the condition. In cured patients, disappearance of the preponderance and persistence of the phase abnormalities are observed. These results suggest a multicanal pathogenesis of Benign Peripheral Paroxysmal Vertigo. PMID- 14526553 TI - Comparison between two perception tests in patients with severe and profoundly severe prelingual sensori-neural deafness. AB - Patients with severe and profoundly severe bilateral sensorineural prelingual deafness constitute a group of particular interest in the organization of the National Health Service; every patient must, in fact, follow a prosthetic rehabilitative-educational programme lasting many years and organized under different areas to compensate for his/her communicative difficulties, especially with regard to the speech canal. No reliable data providing details of the efficacy and efficiency of any of these points is available. A critical point in the rehabilitation process is that of auditory perception training. Of the few auditory perception tests presently available in Italian, the following were examined: namely, EARS (Evaluation of Auditory Responses to Speech) battery, on the one hand, and the Italian version of the ESP (Early Speech Perception), GASP (Glendonald Auditory Screening Procedure). NU-CHIPS (Northwestern University Children's Perception of Speech) and WIPI (Word Intelligibility by Picture Identification) tests on the other. A group of 10 patients presenting severe and profoundly severe bilateral sensori-neural prelingual deafness received the two tests at the beginning and after six months of auditory perception rehabilitation. The findings emerging from the two different test sessions were analysed and compared. The EARS battery was seen to have enabled even very early and highly developed stages of auditory perception to be detected in comparison with the other battery, which, however, was more accurate in evaluating the ability to discriminate and identify words on the basis of their spectral characteristics. The Authors propose the combined use of the two test batteries to evaluate the efficacy and efficiency of auditory perception training in patients with severe and profoundly severe bilateral sensori-neural prelingual deafness. PMID- 14526554 TI - Meniere's disease and gentamicin: preliminary results using the minimum effective dose and integrated therapy. AB - Treatment of Meniere's disease is aimed at restoring a normal quality of life and preserving residual hearing, in view of the increasing frequency with which the contralateral ear is affected. Conventional medical treatment (diuretics + vasoactive drugs) leads to cure in a large percentage of patients (75-95%). In intractable cases, transtympanic (intratympanic) aminoglycoside therapy, associated with various techniques, is becoming widespread as an alternative to surgery. Progressive reduction of the dose introduced into the middle ear did not prevent the onset of anacusis in several patients; the variable, unpredictable permeability of the round window membrane, the object of fundamental studies in the past, explains this complication. The Author has used gentamicin transtympanically in Menierians since 1978, and has treated a total of 105 patients. He first prescribed transtympanic gentamicin therapy that did not follow, but was integrated with conventional medical treatment in 22 intractable Menierians, by instilling the minimum effective dose, to reduce the risk of hearing impairment. Preliminary results, related to stage of disease, may be summarised as follows: improvement in the quality of life, as evaluated by the American Academy of Ophthalmology & Otolaryngology Committee on Hearing and Equilibrium questionnaire (14 patients--63.63%--at point 1 and 8-36.36%--at point 2); disappearance of vertigo in 15 patients (68.18%); a minor vertigo attack in 3 and two minor attacks in 3 others not affecting quality of life; persistence of occasional unsteadiness in one patient. Hearing remained unchanged in 15 patients, improved slightly in 3 cases and worsened slightly in 2; decreased sensitivity to high tones was observed in 2 patients at the first insertion of gentamicin. According to the Author, employing integrated therapy and using the minimum effective dose of gentamicin, the risk of damage to the cochlear structures may be reduced, although not excluded, while restoring a good quality of life, even when repeat instillation is necessary. PMID- 14526555 TI - Development of communication and speech skills after cochlear implant in a sign language child. AB - In selecting patients to undergo cochlear implant, a pre-existing use of sign language gives rise to two problems that have been widely debated in the literature. First, the caution shown toward the candidacy of patients using this mode of communication, since it is considered a possible element of interference in the acquisition of speech. Secondly, refusal of the cochlear implant procedure, on the part of the deaf community, on the grounds both of cultural identity and of it being more "natural" for a deaf person to use an unimpaired visual channel rather than an impaired hearing channel. In order to establish whether knowledge of sign language does, indeed, affect speech production negatively and evaluate which mode of communication, oral or gestual, is preferred, the present investigation was carried out on a preverbal deaf child who had undergone cochlear implant at about 7 years of age and has always used both languages. His verbal skills were evaluated in the precochlear implant stage, then at 6 and 12 months after, together with the changes in his use of sign language and in the relationship between the two modes. Results, besides observing the presence of linguistic evolution at each level examined and already evident at 6 months, also documented a progressive reduction in the spontaneous use of sign language. In conclusion, the present experience revealed no temporal or qualitative differences in post-cochlear implant evolution of speech skills, in comparison with that observed in patients with an exclusively aural-oral approach. Furthermore, the increased use of the hearing pathway, made possible by cochlear implant, determined a spontaneous choice of verbal language as the most natural and economic mode of communication. PMID- 14526556 TI - Localization of the atrial natriuretic factor in the human inferior turbinates. An immunohistochemical study. AB - In man, the architecture of the turbinates is able to modify some of the physiochemical characteristics of the air inhaled. These modifications depend on the nervous system and on the action of neurotransmitters such as vasoactive intestinal peptide. Substance P, calcitonin gene-related peptide and other neuropeptides. As atrial natriuretic factor has been detected in the trachea and lung, the present immunohistochemical study was carried out to establish the presence and localisation of the atrial natriuretic factor on the inferior turbinates of the human being. The findings show atrial natriuretic factor to be present in the serous epithelial cells and in some cells of the tonaca propria near the sinusoids and the arteriovenous shunts and the acinar cells of the glands. Atrial natriuretic factor, therefore, could play a part in the stratification of mucus on the luminal surface and also regulate the blood flow of the capillaries, modifying, in this way, the physiochemical features of the air inhaled. PMID- 14526557 TI - Morphological study of development and functional activity of palatine tonsils in embryonic age. AB - Palatine tonsils play an important role in the development of the immune system, being the first organ in the lymph system which analyses and reacts to antigenic stimulation. In this study, the peritonsillar area of Waldeyer's ring was investigated in 88 normal human embryos which were examined histologically and immunohistochemically. The progressive development of palatine tonsils during embryonic life is discussed. The first appearance of tonsils is in about the 14th 15th week followed by a parallel development of B- and T-cell regions which accounts for the high incidence of non mucosa-associated lymphoid tissue lymphomas among all tonsillar lymphomas and the higher incidence of T-cell lymphomas, in comparison to the mucosa-associated lymphoid tissue of the digestive system. The way in which the human body develops the palatine tonsils quickly and prepares them to react to the first antigenic stimulation, are discussed. PMID- 14526558 TI - Iliac crest free flap in oromandibular reconstruction. 13 cases study. AB - Oromandibular reconstruction following oncological resection is particularly challenging. In this past decade, great improvements have been made, in functional and aesthetic terms, following the introduction of composite free flaps. By means of this reconstructive technique, it is, in fact, possible to restore mandibular continuity, and, consequently, the morphology of the lower third of the face; rehabilitate mastication through osteointegrated implants, and optimise the reconstruction of soft tissue, employing the fascio-cutaneous or muscular portion of these flaps. Between January 1995 and January 2001, 70 oromandibular reconstructions employing osseous free flaps were performed in the Department of Maxillo-Facial Surgery of the Ospedale Maggiore in Parma, and in 13 of these the iliac crest free flap was used. Personal experience is described regarding the indications and use of this flap. PMID- 14526560 TI - Perilymphatic gusher in stapedectomy: demonstration of a fistula of internal auditory canal. AB - Gusher is a very rare phenomenon, generally associated with congenital stapes fixation or otosclerosis in adult age, that may present during stapedectomy. A sudden perilymph flow occurs following platinotomy, due to congenital malformation (abnormally wide cochlear aqueduct or internal auditory canal fistula), that causes an abnormal connection between subarachnoid and perilymphatic spaces. This report deals with a case of bilateral gusher, occurring during stapedectomy, caused by an osseous fistula between bottom of internal auditory canal and the osseous labyrinth later observed at computed tomography scan. The usefulness of a radiologic examination is stressed for a correct therapeutic approach in the even contralateral ear stapedectomy. PMID- 14526559 TI - Centripetal approach to the facial nerve in parotid surgery: personal experience. AB - There is particular interest in parotid surgery on account of the close relationship between the gland and the extrapetrous facial nerve. The seventh cranial nerve is generally located by means of an anterograde or proximal surgical identification technique aimed at identifying the facial nerve at its point of exit from the stylomastoid canal. There are very few reports in the literature on retrograde or centripetal identification techniques, which may be adapted to the morphology of the neoformation limiting surgical access, in order to isolate the nerve from its peripheral rami. The present report deals with personal clinical experience, describing a technique for retrograde detection of the facial nerve. Between 1990 and 2001, 313 parotid surgery procedures were performed at the ORL Clinic of the Universita del Piemonte Orientale in Novara. In 308/313 cases, corresponding to 98% of the operations, the technique chosen for the identification of the extrapetrous facial nerve was anterograde, proximal or centrifugal; in 5 cases alone, retrograde or centripetal exploration of the orbicular branch was undertaken, on account of difficulty in locating the main trunk, due to the presence of a post-inflammatory fibrosis in three patients and a stylomastoid emergency, arising from a malignant neoformation, in the other two. The decision to resort to the identification of the orbicular nerve of the eye is supported by the regular course and adequate size of this facial branch in its peripheral area, which enable it to be easily located. PMID- 14526561 TI - [Medical liability in day-surgery practice in otorhinolaryngology]. PMID- 14526562 TI - [TNM -- classification of malignant tumors (VI edition -- 2002). Innovations in the classification of head and neck neoplasms]. PMID- 14526563 TI - [Do not underestimate infarcts without ST elevation. STEMI or NSTEMI--that is the (infarct) question]. PMID- 14526564 TI - [Educate patients about symptoms! Women with myocardial infarct]. PMID- 14526565 TI - [Guidelines compliant therapy of acute infarct. Primary PTCA is now the first choice]. PMID- 14526566 TI - [Children often suffer unnecessary pain. A fever suppository after surgery is not enough!]. PMID- 14526567 TI - [Subjected to scientific evaluation. What is the value of alternative medicine?]. PMID- 14526568 TI - [On an airplane the day after PTCA? With favorable outcome, there are no contraindications]. PMID- 14526569 TI - [Amalgam, exhaust gases, atomic power--or psychological factors. What is behind environmental anxiety in Germans?]. PMID- 14526570 TI - [Sick building syndrome or fungal allergy? When houses cause illness]. AB - In modern societies, the sick building syndrome (SBS) is a very common building related complex of unspecific symptoms affecting groups of persons. Most frequently, complains include irritation of the eyes and respiratory tract that are believed to be related to negative ambient factors at the workplace. The etiology is multifactorial. In persons showing typical anxiety about the environment, SBS may also be considered a variant of a somatoform disorder. SBS must be clearly differentiated from building-related illness. Diagnostic measures and therapeutic implications are discussed. PMID- 14526571 TI - [Multiple chemical sensitivity. Is the patient suffering as a result of environmental pollutants or psychological problems?]. AB - Multiple chemical sensitivity (MCS) poses a medical challenge. Proposed etiologies are as numerous as they are contradictory, direct and indirect costs are high, and patient suffering considerable. In the absence of objective diagnostic criteria, estimation of its prevalence is difficult. Nevertheless, establishment of the diagnosis is frequently strikingly uncritical. We support an holistic approach that gives consideration both to psychological and physical aspects, as well as taking account of the high level of comorbidity, and we warn against "over-diagnosis". Therapeutical approaches should consider carefully the risk of avoidance and social withdrawal. PMID- 14526572 TI - [Ill due to amalgam? 10 rules for managing the symptomatic patient]. AB - Over the past two decades, mercury released by amalgam fillings has been held responsible for a number of mental and somatic health complaints. However, a systematic relation between increased mercury levels and the severity of the reported symptoms has never been demonstrated in any of the present well controlled multidisciplinary studies. These studies, however, have found a high prevalence of mental disorders, especially somatization syndromes, among patients with self-diagnosed "amalgam illness". Additionally, our own studies indicate that amalgam anxiety is often merely one aspect of a general environmental anxiety. Overall, the present findings suggest a psychological etiology for amalgam-related complaints. Our psychosomatic model of "amalgam illness" integrates external factors, individual predispositions and specific processes of perception, awareness, evaluation and attribution. Practical management strategies for primary care physicians can be derived from this model. PMID- 14526573 TI - [Acute dyspnea in childhood. Croup or foreign body--bronchiolitis or asthma?]. AB - The diagnosis and treatment of acute respiratory distress or dyspnea in children needs to consider the age-related major causes, but also the rare causes. In neonates, congenital abnormalities, immaturity of the lungs, perinatal complications and infections are major causes, in infants bronchiolitis/obstructive bronchitis and croup, while in older children and adolescents acute asthmatic attacks and pneumonia predominate. Of critical importance is a clear diagnostic classification, assessment of severity, securement of adequate oxygenation and a carefully taken decision for hospital or ambulatory treatment. Where indicated, less invasive therapeutic options (inhaled bronchodilators, rectal or oral steroids) are to be preferred over more invasive (i.m., i.v.) alternative measures. PMID- 14526574 TI - [3 basic symptoms: affective--somatic--cognitive. Recognizing the faces of depression]. PMID- 14526575 TI - [Antihypertensive drugs for dementia?]. PMID- 14526576 TI - [Helicobacter eradication in gastritis and ulcers. Causal therapy, prevention of stomach cancer]. PMID- 14526577 TI - [Helicobacter pylori eradication in ulcer patients. Quality of life improves, costs decline]. PMID- 14526578 TI - [Double profit for hypertensive patients. Blood pressure decreases--frequency of coitus increases]. PMID- 14526579 TI - [Less pain after surgical intervention. Starting analgesia already before surgery]. PMID- 14526580 TI - [Thiazide analogs. Also can be used in renal failure]. PMID- 14526581 TI - [No reimbursement for non-prescription medicine. Expensive savings plan?]. PMID- 14526582 TI - [Lipidocolloid dressing can be changed without wound damage. No adhesion to the wound]. PMID- 14526584 TI - [Antibodies, antihypertensive drugs, antioxidants. Can they prevent Alzheimer disease?]. PMID- 14526583 TI - [Fasting hyperglycemia and postprandial glucose peaks. Diabetes therapy mustact on 2 sides]. PMID- 14526585 TI - [Most hypertensive patients need drug combination treatment. "First line therapy is in second place"]. PMID- 14526586 TI - [New protease inhibitor against HIV. Viral load decreases--without lipid increase]. PMID- 14526587 TI - [Acute back pain. Two-thirds are functional complaints]. PMID- 14526588 TI - [Dyspnea. When is it pulmonary hypertension?]. PMID- 14526589 TI - [How does it work? Skeletal scintigraphy]. PMID- 14526590 TI - [Diagnosis at a glance. What clouds the patient's vision?]. PMID- 14526591 TI - Female offenders in a changing society. PMID- 14526592 TI - Female repeat offenders negotiating identity. AB - This study examines the process of identity negotiation for Israeli female ex convicts who were separated for extensive periods of time from their children and eventually lost custody over them. The content analysis of in-depth interviews reveals that these women were able to reconstruct their biographies and retrospectively account for their crimes and drug addiction in terms of the sexual, physical, and economic abuse they had endured and by appeal to higher loyalties, their children who they had to provide for. However, when having to account for their fallings as mothers, all biographical reconstruction, external blame, and accusation collapsed. Looking at themselves through their children's eyes, female offenders were simply unable to renegotiate the imputed identity of incompetent mother. They could neither confront their children's anger nor explain to them why they had abandoned them. Permanently alienated from the center of motherhood, these women were doomed to an existential chaos. PMID- 14526593 TI - The effectiveness of relapse prevention with offenders: a meta-analysis. AB - Although relapse prevention models have been applied within offender treatment, there has been little controlled outcome research evaluating their effectiveness. This meta-analysis of 40 tests of relapse prevention treatment revealed moderate mean reductions in recidivism (0.15), and certain elements of the relapse prevention model (i.e., training significant others in the program model and identifying the offense chain) yielded stronger effects than others (i.e., provision of booster/aftercare sessions and developing coping skills). Further analyses revealed that the clinically relevant and psychologically informed principles of risk, need, and general responsivity yielded the strongest reductions in recidivism. The implications for future research and treatment are discussed. PMID- 14526594 TI - Familial functioning as a support system for adolescents' postdetention success. AB - Youths transitioning out of detention are particularly in need of support systems that can divert them from recidivism. This article examines the relationship between incarcerated youths' sense of their families' functioning and their perceived likelihood for postdetention success. Despite the control of background variables, results reveal that delinquents who indicated stronger familial functioning also reported a greater likelihood of future success. Although interventions that focus on improving the supportive capacity of families have shown great promise, treatment drop-out rates remain high. To explore incarcerated youths' view of family focused interventions, respondents rated the potential utility of family and nonfamily focused intervention options. A majority of respondents reported that family focused interventions would be helpful. A group of respondents, overrepresented by adolescents with low family functioning, reported that this type of intervention would not be helpful. These results are discussed in terms of their impact on offering family focused interventions for juveniles. PMID- 14526595 TI - An exploration of childhood victimization and family-of-origin characteristics of sexual offenders against children. AB - A comparison of the childhood victimization experiences and family-of-origin characteristics of 33 sexual offenders against children, 66 violent offenders, and 25 nonviolent offenders participating in assessment or treatment at a forensic division of a Canadian psychiatric hospital was undertaken using a semistructured interview designed specifically for this study. Overall, 31.5% of the sample reported contact sexual abuse (i.e., oral, vaginal, or anal) by age 14. Sexual offenders against children reported significantly more sexual abuse than both violent and nonviolent offenders and were more likely to have been sexually propositioned and exposed to. Sexual offenders against children were also more likely to report physical discipline as the primary type of discipline in the home. Analyses of the variables concerning characteristics of parents, siblings, and significant others also indicated significant differences between groups. Implications of these findings on the intergenerational hypothesis are discussed as well as their relevance for treatment. PMID- 14526597 TI - Eastern Turkistan Islamic Movement: a case study of a new terrorist organization in China. AB - The Eastern Turkistan Islamic Movement (ETIM) was designated a terrorist organization by Afghanistan, Kyrgyzstan, China, the United States, and the United Nations in 2002. However, no systematic studies have been published on the new terrorist organization in Xinjiang, China. Using a case-study approach and interviews, this article attempts to provide information in terms of its historical evaluation, related religious and ethnic issues, organizational agenda, activities, and role in the current international terrorist network. This article argues that better international cooperation and the improvement of social and religious policies will help curtail activities of the ETIM. PMID- 14526596 TI - Treating impulsivity, irritability, and aggression of antisocial personality disorder with quetiapine. AB - There are few effective treatments of antisocial personality disorder (APD). Preliminary work suggests that the atypical antipsychotic quetiapine can decrease irritability, impulsivity, and aggressiveness. Data were collected from 4 patients with APD who were referred to a maximum-security inpatient psychiatric facility for pretrial evaluation and were treated with quetiapine. Quetiapine was effective in these patients as was indicated by a decrease in symptoms such as impulsivity, hostility, aggressiveness, irritability, and rage reactions. Typical dosage was 600 to 800 mg per day. Patients attributed their willingness to comply with quetiapine treatment to both the effectiveness of the drug and its favorable adverse-event profile. Quetiapine was successfully combined with mood stabilizers, particularly gabapentin, in patients with prominent affective instability. Quetiapine has demonstrated efficacy in aggression, impulsivity, and irritability and has proved to be an effective medication in these patients with APD. In addition, its favorable adverse-event profile makes patients willing to comply. PMID- 14526598 TI - Comparability of the MMPI-2 F(p) and F scales and the SIRS in clinical use with suspected malingerers. AB - Relationships among Structured Interview of Reported Symptoms (SIRS) scores and Minnesota Multiphasic Personality Inventory--2 (MMPI-2) F(p) and F scores were examined for 63 suspected malingerers evaluated at either of two psychiatric facilities. Despite differences between facilities in terms of seriousness of subjects' offenses, mean scores on the malingering tests were similar. Cutting scores for F(p) and F resulting in substantial correspondence between these scales and the SIRS were derived. Use of the cut score for F(p) proposed by Arbisi and Ben-Porath (1995) resulted in less agreement with the SIRS than did a lower cut score. No substantial difference between F(p) and F in each scale's overall agreement with the SIRS was observed. A principal components analysis of the SIRS primary scales produced two factors, interpreted as Overreporting of Symptoms and Implausible Symptoms. F(p) was observed to correlate significantly with Implausible Symptoms but not with Overreporting of Symptoms; F was significantly correlated with both factors. PMID- 14526599 TI - A program analysis of Esuba: helping turn abuse around for inmates. AB - This investigation is an empirical evaluation of Esuba, an abuse awareness program implemented in the Florida correctional system. Research has indicated a link between violent and abusive behavior and criminal activity. This study specifically addressed whether the Esuba program had an impact on offenders' self esteem, stability of self, faith in people, and sensitivity to criticism as measured by the Rosenberg Self-Esteem Scale. The results from the application of the Esuba program to an incarcerated offender population are encouraging. The results suggest that the Esuba program does appear to significantly increase offenders' emotional self-preservation. PMID- 14526600 TI - [On monitoring chemicals]. AB - The authors necessitate biologic monitoring to be conducted along with chemical monitoring of environmental objects. Accomplishment of biologic monitoring and interpretation of its results are associated with difficulties. Being a valuable addition to chemical monitoring, biologic monitoring is a total chemical dose incorporated by a biologic object. PMID- 14526601 TI - [On groups maximal allowable concentration for nickel and its inorganic compounds]. AB - The article deals with setting a unified group Maximal Allowable Concentration for inorganic nickel compounds that pollute ambient air in hydro- and pyrometallurgic industries. PMID- 14526602 TI - [Occupational risk and its prophylaxis for female workers engaged in radio electronic instrument industry]. AB - Hygienic evaluation of work conditions and health state of women engaged into radioelectronic instrument-making industry helped to define occupational hazards determining occupational risk that includes criteria for reproductive disorders. PMID- 14526603 TI - [Hygienic evaluation of work condition and health status of female telephone operators in information services]. AB - The authors conducted hygienic evaluation of work conditions for female telephone operators of inquiry office. The article deals with characteristics of health state according to periodic medical examinations and with evaluation of the workers' reproductivity according to poll data. The analysis covered influence of sensory and emotional load on health state of female telephone operators. PMID- 14526604 TI - [Using pulse measurements for physiological regulation of work intensity]. AB - The article covers materials of physiologic evaluation of work intensity through pulse measurements. The authors stress drawbacks of present norms for average shift pulse rate. To evaluate work intensity, the researchers suggest normal value of pulse rate change at the 7th work hour vs at the 2nd hour. PMID- 14526605 TI - [Hygienic evaluation of work conditions at IR-lasers production]. AB - Work conditions at IR-lasers production are associated with exposure to a complex of hazards including laser irradiation, psychoemotional strain, visual strain. PMID- 14526606 TI - [Ecological and hygienic studies of electromagnetic irradiation of navigation safety system in Eastern area of the Finnish Gulf]. AB - The authors evaluated electromagnetic situation near radiotechnical emitters on 8 objects of regional navigation safety system located in St. Petersburg Seaport Administration. The results proved that exploitation of the radiotechnical means does not result in electromagnetic irradiation that exceeds maximal allowable levels. PMID- 14526608 TI - [Preventing hand disorders in metallic hulls assemble workers]. PMID- 14526607 TI - [Joint and vertebral pain in workers of aluminum production]. PMID- 14526609 TI - [Features of somatic diseases in electric welders with chronic manganese intoxication]. PMID- 14526610 TI - [Diagnostic and prognostic value of serum manganese levels for chronic manganese intoxication in electric welders]. PMID- 14526611 TI - [Mechanism underlying the influence of infrasound on humans]. PMID- 14526612 TI - [Comet or snoop?]. PMID- 14526613 TI - [Fusion proteins in HIV-AIDS treatment]. PMID- 14526614 TI - [Radiosynovectomy. Nuclear medical treatment for inflammatory rheumatic joint diseases]. PMID- 14526615 TI - [Boswellic acids (the active agent in incense) for the treatment of chronic inflammatory diseases]. PMID- 14526616 TI - [Inflammation in mouth and throat]. PMID- 14526617 TI - [Not every TIA is primarily vascular]. AB - Three patients, two women aged 72 and 45 years, and a man aged 80 years, presented with transient neurological deficits due to a brain tumour, a glioblastoma multiforme and two meningiomas respectively. A fourth patient, an 84 year-old man, had a transient ischaemic attack (TIA) with a meningioma as an incidental finding. The first woman had normal CT findings, but MRI revealed the neoplasm. Symptoms included motor loss, sensory disturbances, dysphasia and dysarthria, lasting from 30 seconds up to 10 minutes. The first two patients had surgery; the first one later died when the tumour recurred. The other two patients still exhibit a spontaneous recovery. Of all patients with a clinical presentation of a TIA, 0.4-1% harbour a brain tumour. Clinical symptoms do not distinguish 'transient tumour attacks' from TIAs with a primarily vascular origin. Transient tumour attacks are mainly seen with meningiomas, and to a lesser extent with high-grade gliomas. Changes in intracranial pressure leading to focal ischaemia may explain the occurrence of this phenomenon. A part from intracerebral tumours, non-vascular entities mimicking TIAs can also be seen with demyelinating processes, metabolic disturbances, epilepsy or migraine. Brain imaging is always required in patients with transient neurological deficits. A CT scan may provide false-negative results and in case of doubt, MRI is the preferred diagnostic tool. PMID- 14526618 TI - [The termination of life in the Netherlands and in Europe]. AB - Medical decisions concerning the end of life, including the termination of life, are taken not only in the Netherlands but in other European countries as well. In some of these countries, the social pressure to reform legislation is increasing. The World Medical Association remains strongly opposed to any involvement of physicians in bringing about people's death, but the rulings of the European Court of Human Rights so far do not prohibit national laws to regulate euthanasia. Whilst in other countries there is debate on whether permissive legislation should be adopted, in the Netherlands the emphasis is now on the development of instruments aimed at the quality and transparency of medical practice as well as on the improvement of the notification procedure, reducing the gap between medical and juridical reality. PMID- 14526619 TI - [Monitoring and treatment of patients with shock in the intensive care unit]. AB - The controversy surrounding the use of the pulmonary artery catheter, has stimulated research into alternative methods of haemodynamic monitoring. As yet, however, no new gold standard has emerged. In the future, interest in haemodynamic monitoring is likely to focus more on tissue perfusion and metabolism instead of central circulation. Important causes of shock in the ICU, apart from acute blood loss, are sepsis and acute heart failure. Septic shock results from vasodilatation and myocardial dysfunction. Early initiation of aggressive fluid resuscitation, if necessary accompanied by vasoactive and inotropic agents, improves survival. In addition, low dose corticosteroids have a positive impact on mortality. In the treatment of patients with acute heart failure, phosphodiesterase III-inhibitors are becoming part of standard therapy in addition to beta-adrenoceptor agonists, especially in patients who take beta blockers. PMID- 14526620 TI - [Clinical reasoning and decision making in practice. A 41-year old with periodic fever of unknown origin]. AB - A 41-year-old man presented with unexplained bleeding from the right tonsil. He subsequently developed periodic fever, cervical lymphadenopathy and hepatosplenomegaly. Despite extensive bacteriological, serological and radiographic investigations for infectious disease, rheumatic disease and malignancy no diagnosis was made. Although the fever pattern was very suggestive of Pel-Ebstein fever--commonly associated with lymphoproliferative disease- multiple biopsies of lymph nodes, bone marrow, tonsils and liver all proved negative. Empirical glucocorticoid therapy gave some temporary improvement lasting for a month. Splenectomy or splenic biopsy was not carried out because of the risk of excessive bleeding. Eventually the patient died of multi-organ failure and sepsis. At autopsy, a T-cell lymphoma with an unusual phenotype and focal involvement of bone marrow, liver and spleen was found. Clinicians are sometimes faced with the dilemma of whether to perform multiple, invasive and possibly harmful diagnostic tests or to start empirical therapy. Empirical therapy may only be started if the diagnosis has been made on strong clinical grounds and, if this is not the case, only after further diagnostic tests. The question of whether a potentially harmful diagnostic test is justified depends on the clinical course, the sensitivity and specificity of the test and the therapeutic possibilities. PMID- 14526621 TI - [Diagnostic image (156). A man with a hematoma behind the ear. Mastoid bone fracture]. AB - After a fall against a concrete ridge, a 65-year-old man was seen at the first aid department with a haematoma behind his right ear (Battle's sign) and blood loss from the external meatus. A CT scan showed a basal skull fracture located in the right mastoid bone. PMID- 14526622 TI - [Acquired von Willebrand syndrome]. AB - Two patients, a man aged 69 and a woman aged 64, were diagnosed with Von Willebrand syndrome caused by monoclonal gammopathy. The man, who was admitted for hip surgery, had a history of long episodes of epistaxis. The patient was treated with immunoglobulin and the hip operation was carried out with no complications. The woman suffered from haemorrhagic diathesis. She was advised that should she undergo an invasive procedure then treatment with a prophylactic with intravenous immunoglobulin or Von-Willebrand factor (VWF)/factor-VIII concentrates must be administered. Acquired Von-Willebrand syndrome is a rare condition with an estimated prevalence of 0.04-0.13%. It is linked to a large number of underlying diseases such as paraproteinaemia, multiple myeloma (Kahler's disease), myeloproliferative disease, lymphoproliferative disease, auto immune disease, solid tumours and hypothyroidism. Recognition depends on a careful case-history and identification of the underlying disease. For its diagnosis VWF antigen. VWF propeptide, activated partial thromboplastin time and factor VIII are of importance. Technically, it is difficult to show the presence of VWF antibodies as it concerns a heterogeneous group of antibodies. There are two pillars of treatment: symptomatic treatment of the bleeding tendencies using desmopressin, VWF-concentrate or intravenous gammaglobulin, and treatment of the underlying disease. The latter form of treatment can lead to acquired Von Willebrand-syndrome disappearing altogether. PMID- 14526623 TI - [Issues involved in Dutch general practitioners' failure to request autopsy: report from three focus groups]. PMID- 14526624 TI - [Issues involved in Dutch general practitioners' failure to request autopsy: report from three focus groups]. PMID- 14526625 TI - [Issues involved in Dutch general practitioners' failure to request autopsy: report from three focus groups]. PMID- 14526626 TI - [A case of fraud in a neurological pharmaceutical clinical trial]. PMID- 14526627 TI - [Neurosyphilis in psychiatry: surprising underlying disorder in 3 patients]. PMID- 14526628 TI - [Complications of liver cirrhosis: portal hypertension, gastroesophageal varices and ascites]. AB - Patients with cirrhosis of the liver are at high risk of a large variety of complications. Especially the development of portal hypertension, followed by gastroesophageal varicosis and ascites are potentially life threatening problems. In the treatment of gastroesophageal varicosis primary prophylaxis to prevent a first bleeding episode, acute therapy for bleeding varices, and secondary prophylaxis to prevent patients from rebleeding have to be considered. While treating patients with ascites the high frequency of side effects induced by diuretics has to be taken into account. In addition, the diagnosis of spontaneous bacterial peritonitis must not be missed. Hepatorenal syndrome, a typical complication of advanced cirrhosis is especially difficult to treat and is considered an indication for liver transplantation. PMID- 14526629 TI - [Imaging in gastroenterology--what is new?]. AB - In the last years there are a lot of developments in endoscopical, radiological and sonographical imaging techniques for example virtual colonoscopy, capsule endoscopy and contrast-enhanced sonography. The MR- or CT-colonography has a high sensitivity of about 90% in the detection of polyps > 1 cm. But what there is to be said against virtual colonoscopy as a routine examination is the need for bowel cleansing with an oral lavage and the high costs due to a long examination and evaluation time. The capsule endoscopy is a very interesting new imaging technique for the diagnostic evaluation of small bowel diseases. The capsule endoscopy is superior to push-enteroscopy, the gold-standard until now. But it is not clear, whether there is change in therapeutical procedures due to the results of the capsule endoscopy and whether the high costs for the examination are justified. The contrast-enhanced sonography improves especially the detection and characterization of solid liver lesions and some CT- and MR-examination will be unnecessary in the future. PMID- 14526630 TI - [Cutaneous neoplasms]. AB - The skin is the organ most commonly affected by malignancies. Various cancers of the skin show a dramatic increase in incidence over the last decades. Epithelial skin tumors are most frequently, e.g., basal cell carcinoma and the squamous cell carcinoma with its precursors, the actinic keratoses. Melanoma, which is extremely difficult to treat in advanced tumor stages, is dreaded. Besides that, there are other epithelial malignant diseases, e.g. Morbus Bowen and adnexal tumors originating from the skin appendices. Mesenchymal malignant neoplasias such as Morbus Kaposi, angiosarcomas and other dermal sarcomas, are rare. Since the majority of malignant neoplasms is removable and curable by a simple surgical intervention, the knowledge of the different skin tumors is essential for non dermatologist. PMID- 14526631 TI - [Cutaneous manifestations of internal diseases: purpura, livedo, pyoderma gangrenosum]. AB - Dermatology and internal medicine have in common that many systemic diseases manifest with skin symptoms that are easily accessible for both diagnostic and investigative procedures. Even if modern practice of medicine tends towards organ specific particularity of specialists, dermatology traditionally strives for interdisciplinary communication. Cutaneous manifestations of internal diseases present in various forms. In a syndrome, a number of symptoms present in an identical pattern, and pattern recognition is diagnostic. Vice versa, a distinct skin sign can be a clue to various internal disorders, that have to be considered in the differential diagnosis. Purpura and livedo due to either inflammatory (immune complex mediated or associated with ANCA) or vasoocclusive microthrombotic vascular disease are presented as examples. Finally, subtile variations in key symptoms of the skin, such as of pyoderma gangrenosum, may give a clue to a specific underlying disease, such as rheumatoid arthritis, inflammatory bowel disease, monoclonal gammopathy, or hematologic disease, as exemplified by the variants of pyoderma gangrenosum. PMID- 14526632 TI - [Hair loss]. AB - Hair loss includes excessive shedding of hair (effluvium) and the alopecias, in which there is a decrease in the amount of hair. Both may occur diffusely, or in a circumscribed manner. The localization and pattern of circumscribed hair loss may give a clue to its cause. On the basis of morphologic criteria the alopecias are further classified into non-scarring and scarring types. Non-scarring alopecias (diffuse alopecia, androgenetic alopecia, alopecia areata) are essentially hair cycling disorders, while scarring alopecias result from irreversible destruction of the hair follicles with recognizable loss of follicular osita. Scalp biopsy should be an early step in the evaluation of any case of alopecia, in which scarring is suspected. Too little scalp hair is not a vital problem, but represents a major health care challenge. Only recently the psychological impact of hair loss has been appreciated by the medical community, though throughout history, too little hair has been a concern to mankind, and the object of cosmetic interest and quackery. Recent advances of the medical sciences have led to a better understanding of the underlying pathogenic processes and opened the venue to effective pharmacotherapy (minoxidil, finasteride) and technologies (autologous hair transplantation) for the treatment of the most common disorders. Together with the availability of such treatments, high technical standards for evaluating their efficacy have been developed and have become mandatory for appraisal of any hair growth promoting agent, both in clinical studies and in the individual patient. PMID- 14526633 TI - Limping in childhood. AB - Limp is an abnormal gait which can be caused by pain, muscular dysfunction or deformity. Limp is never normal and the cause should be established. There is a long list of possible diagnoses. A complete history and thorough physical examination are the most helpful tools in sorting out the various causes of limping. Laboratory tests and imaging studies should be based on findings in the history and physical examination. Certain causes of limping must be diagnosed at the first visit. Sometimes the cause of limping cannot be determined, and after exclusion of any serious disease, the child should be observed. Many of them will have spontaneous resolution without treatment. PMID- 14526634 TI - Vascularised pisiform bone graft. Indications, technique and long-term results. AB - The authors report their experience with the use of a vascularised pisiform bone graft based on the dorsal branch of the ulnar artery to provide osseous support and an efficient vascular aid especially in non-unions of the carpal navicular bone with avascular necrosis of its proximal pole and in stage III Kienbock's disease. The pisiform can replace the proximal pole of the navicular bone in totality. When implanted into the lunate, it stops its collapse and helps to revascularize it. The authors present the results achieved in 14 patients (12 male, 2 female), of which eight had a follow-up longer than 5 years The technique appears as an interesting alternative to carpectomies and partial intracarpal fusions which are usually proposed in advanced cases of these conditions. PMID- 14526635 TI - Carpal coalition: a rare coincidence with hand deficiencies. AB - Three cases in which congenital carpal fusion was associated with formation deficiencies of the hand are presented. In two patients with a lunate-triquetral and a capitate-hamate-trapezoid coalition, respectively, an ulnar deficient hand was also diagnosed. A subdivision of the first type of ulnar club hand deformities is proposed. Other skeletal anomalies included duplication of a phalanx and pseudoepiphysis of a metacarpal, respectively. In the latter case hypoplasia of the flexor tendons to the fingers, excluding the thumb, was also noted. In the third case, a proximal capitate-hamate coalition coexisted with aplasia of the flexor tendons to the middle finger. PMID- 14526637 TI - Influence of operation duration on perioperative morbidity in revision total hip arthroplasty. AB - This retrospective study investigated the impact of patient and procedure-related parameters on the complication rate following revision total hip arthroplasty. Complications included vessel and nerve damage, periprosthetic femoral fracture, wound infection, wound bleeding, prosthesis dislocations, thromboembolism, cardiac and pulmonary complications, and death. The influence of operation duration, gender, revision status, ASA classification, and type of fixation of the primary implant on the perioperative morbidity was investigated in a sample of 60 revision procedures (cemented stems, cemented or cementless cups). Odds ratio [OR] and 95% confidence interval [CI] were estimated with multiple regression models. Perioperative morbidity was significantly correlated to operation duration (OR = 1.03; CI: 1.00-1.05), but not to age (OR = 1.01; CI: 0.93-1.09), gender (OR = 2.66; CI: 0.50-14.05), revision status (OR = 2.34; CI: 0.54-10.05), ASA classification (OR = 1.24; CI: 0.30-5.18), or type of fixation of the primary implant (OR = 2.49; CI: 0.47-13.17) Duration of the revision operation appeared as a predictive parameter for perioperative morbidity in revision total hip arthroplasty in our study group. PMID- 14526636 TI - The value of external fixation for unstable pelvic ring injuries. AB - Unstable pelvic ring injuries are severe injuries, with high morbidity and mortality rates. Since 1994, the authors have been using external fixation to treat these injuries. They present the results achieved in a series of 22 cases of unstable pelvic ring injuries admitted to their institution between 1994 and 2001. External fixation was performed in all cases with pins inserted between the anterior superior and the anterior inferior iliac spine. Posterior injuries were treated operatively in four cases and non-operatively in others. The average age of the patients was 32 years (range: 22 to 56 years). According to Tile's classification, there were 10 type B and 12 type C injuries. Functional evaluation using Majeed's score at the time of review showed a mean score of 75.6 (range: 12-100). The pain score was 22.7/30. Radiological results were good in horizontal injuries, with satisfactory reduction of the symphyseal disruption. In vertical shear injuries however, skeletal traction alone did not achieve accurate posterior reduction in four cases, in which internal fixation was required. Anteroinferior placement of external fixation pins appears as a safe technique with a potential for increased stability of fixation. Associated posterior injuries must be treated with direct internal fixation in cases with vertical instability in which heavy skeletal traction has failed to achieve reduction. PMID- 14526638 TI - Does harvesting the medial third of the patellar tendon cause lateral shift of the patella after ACL reconstruction? AB - Many centers do not use the medial third of the patellar tendon as a graft for ACL reconstruction due to the apprehension that there may be post harvest maltracking of the patella towards the lateral side. We undertook a prospective study to evaluate patellar alignment in 30 patients in whom ACL reconstruction was done using the medial third of the patellar tendon. Q-angle assessment was the rough clinical parameter, and the Merchant view, Laurin views, and CT scans were used for radiological assessment. The assessment was done preoperatively and at six months post surgery, and the contralateral normal knees were used as controls. We observed no significant changes in the congruence angle (pre-op 13.86 degrees, postop -12 degrees) or the lateral patellofemoral angles in the pre-operative and postoperative knees as compared to the controls. CT scans were more sensitive for evaluating the congruence angles, and picked up an insignificant abnormal lateralisation of the patella in four cases. None of these patients however had any postoperative symptoms of misalignment. Although medial third graft harvesting did have a 13% change in patellar alignment, this was not statistically significant. Post harvesting local problems are inherent to the procedure, and are independent of the site of harvesting, as they were also seen in cases with no evidence of misalignment. PMID- 14526640 TI - Medium to long-term functional outcome of patients after chemonucleolysis. AB - Chemonucleolysis is an established modality in the treatment of lumbar disc prolapse and has been widely used for over 39 years since its introduction by Lyman Smith in 1963. We report the medium to long-term functional outcome of patients who had chemonucleolysis for single level disc prolapse. One hundred and twelve patients were reviewed retrospectively with a mean follow-up of 9.5 years. The Oswestry Disability Index questionnaire was used to estimate the functional outcome of chemonucleolysis. An excellent or good response occurred in 79 patients (70.5%) while 12 patients (10.7%) showed moderate response with minimal disability. Treatment failed in 21 patients (18.5%) who showed poor response and 12 of these 21 patients went for surgery within a mean period of 6 months. One patient had surgery at a different level than chemonucleolysis. There was only one incident of procedure termination because of epidural contrast leak. There was no case of anaphylaxis or discitis. We concluded on the basis of our results that in carefully selected patients, chemonucleolysis is a safe and effective treatment modality for lumbar disc herniation with good medium to long-term functional outcome. PMID- 14526639 TI - Isolated meniscal repair in the avascular area. AB - The authors present the results of a prospective cohort study carried out in young patients, to evaluate the outcome of meniscal repair in the avascular region of the meniscus. Forty five patients underwent either open (23 patients) or arthroscopic repair (22 patients) between 1982 and 1992. Seventeen men and eight women were included in the present study; their ages at the time of admission ranged from 16 to 27 years. They all presented with isolated longitudinal tears in the central avascular region of the meniscus. No patient with any kind of concomitant lesion was included in this series. The stitches were usually passed through the injured area and the synovium, and the knots were tightened outside the capsule. Twenty five patients of the total series, were subjected to a clinical examination in 1993, 9 to 54 months after meniscal suture. The Lysholm Knee Rating Score and the Tegner Activity Test were used. During 1999, these patients responded to a written questionnaire, based on the above mentioned rating scales. Among the 25 patients, there were 4 recurrences (16%), 2 fair results (8%) and one partial healing (4%). Eighteen (72%) patients, among which eight were treated using the arthroscopic technique, have shown favourable results, without any symptoms from the tibiofemoral joint, based on clinical examination and on the Lysholm Rating Scale and the Tegner Activity Test. When confronted with meniscal tears where there is doubt regarding the vascularity of the injured area, surgical repair in the avascular zone appears to provide a reasonable probability of healing, with good clinical outcome. PMID- 14526641 TI - Results of surgical treatment of osteoid osteoma of the spine. AB - Ten percent of all osteoid osteomas are located in the spine; they represent the most common cause of painful scoliosis in adolescents. The recommended treatment is surgical excision of the nidus. Nine patients with osteoid osteoma of the spine underwent surgical treatment between 1996 and 2002. Complete nidus removal was performed in all patients and the diagnosis was made by frozen sections intraoperatively. The follow-up period was 27 months. Scoliosis was present in five of the cases (55.5%). All deformities improved during the postoperative follow-up period. All nine patients with the diagnosis of osteoid osteoma who had operations had histological confirmation. All patients had immediate and complete relief of pain after surgery. There were no complications and no local recurrences after a follow-up of two years. In the authors' opinion, removal of the nidus by open surgical technique remains a valid method of treatment for osteoid osteoma of the spine. PMID- 14526642 TI - Implementation of a CT-based navigation system in two-stage reimplantation for infected total knee arthroplasty. AB - Femorotibial alignment is an important factor affecting patient outcome after total knee arthroplasty (TKA). It was the aim of this study to report our first results using a CT-based navigation system in two-stage revision surgery for infected TKA. Two patients with chronic deep infection after primary TKA underwent two-stage revision arthroplasty with temporary articulating cement spacers followed by prosthesis re-implantation using a CT-based navigation system. Postoperative radiographs showed accurate alignment of the femoral and tibial components. CT-based navigation systems offer the opportunity for preoperative planning and accurate intra-operative navigation of cutting blocks. They can be considered of value for primary TKA but even more so for two-stage revision arthroplasty. PMID- 14526643 TI - Kienbock's disease in a 12-year-old girl. AB - A case of avascular necrosis of the lunate in a 12-year-old girl is described. Conservative treatment resulted in a clinically normal wrist at three years' follow-up, despite obvious radiological changes. PMID- 14526644 TI - Giant-cell reparative granuloma of the tibia. AB - Giant-cell reparative granuloma (GCRG) occurs in the jaw, temporal bone, and short tubular bones of the hands and feet. Although GCRG can affect long bones, only small numbers of such cases have been sporadically reported. This report describes a giant-cell reparative granuloma in the proximal tibia in a 60-year old woman, describes features of GCRG in long bones and reviews the literature. A 60-year-old female patient was referred to us with complaints of moderately tender swelling of the right leg. Whole-body scintigraphic scanning was performed, which incidentally also disclosed a distal femoral lesion. The patient was admitted for surgery and incisional biopsies were performed on both lesions. Pathology analysis of the specimen from the tibia showed new bone lamellae encircled by osteoblasts and multinucleated giant cells which were more numerous in the haemorrhagic regions of the stroma; the latter displayed fibroblasts, histiocytes and inflammatory cells. The specimen from the femoral lesion showed typical features of a benign enchondroma. The patient was readmitted for surgery. The femoral enchondroma was curetted and the cavity was packed with bone graft. The tibial GCRG was treated with marginal resection, autogenous and allogenous bone grafting and intramedullary nailing. Follow-up examination after two years showed no clinical or radiological evidence of a recurrence. Although GCRG is uncommon, it should be considered whenever a lucent, expansile, and possibly destructive lesion of a long bone is encountered. It should be distinguished from true giant cell tumours occurring in the same locations because they have different biologic behaviours. PMID- 14526645 TI - Tumoral calcinosis with unusual presentation. A case report. AB - A case of multiple para-articular calcific masses involving the elbow, hand, back, knee and the feet was encountered. Pathology after surgical excision of all the masses revealed tumoral calcinosis. The unusual feature was the huge size of the lumbar and knee masses, and the distal location of the swellings in the hand and the foot; there was no disturbance of calcium and phosphorus metabolism, which was an unusual feature of the condition in this patient. PMID- 14526646 TI - Non-traumatic shoulder instability in an athletic patient with a periglenoidal cyst and a glenoid labral tear. AB - A 34-year-old patient presented to our outpatient clinic with the chief complaint of shoulder instability, without any history of trauma. Physical examination revealed a painful apprehension test at 60 degrees, 90 degrees and 120 degrees but no objective sign of shoulder instability or hyperlaxity. MRI-scan showed a cyst over the anterior inferior glenoid rim. Arthroscopic findings were an enlarged capsule, a positive drive-through sign, a SLAP I lesion and a sublabral cyst at the anterior-inferior labrum. Detachment of the anterior labrum could be detected with a probe. The cyst's membrane was resected using a whisker shaver. The capsule and the anterior labrum were refixated with a suture anchor. Following capsular shrinking, there was no further laxity and the drive-through sign was diminished. After three months there was full range of active and passive motion. The patient had no subjective instability sensations. MRI showed no residuum of the cyst. Juxta-articular cysts are a known entity in large joints. There are different types of periarticular cysts. A ganglion cyst of the shoulder associated with glenohumeral instability has, to our knowledge, only been described twice. Our case suggests that mere excision of a juxtaglenoidal ganglion is not sufficient; reconstruction of the labrum must be performed to restore stability of the shoulder. PMID- 14526647 TI - Distal femoral cortical irregularity in a 13-year old boy. A case report. AB - Distal femoral cortical irregularity (DFCI) is a benign entity presenting with either an irregular appearance or a focal radiolucency within the posterior cortex of the distal femur. Atypical cases should be differentiated from malignant lesions. The typical location of the lesion at the attachment of the head of the medial gastrocnemius muscle can be visualised using different imaging techniques. We report a case of a DFCI in a 13-year old boy. The diagnosis was based on CT-imaging. Two years later, no radiological abnormalities could be observed. A review of the literature is presented. PMID- 14526648 TI - Pseudoaneurysm complicating knee arthroscopy. A case report. AB - Pseudoaneurysm formation is a rare complication after knee arthroscopy. Most pseudoaneurysms complicating knee arthroscopy develop on the popliteal and geniculate arteries. We report a case of a pseudoaneurysm developed on an arterial branch to the medial gastrocnemius muscle, as a complication of an arthroscopic meniscectomy in a 66-year-old patient. The patient was treated successfully by resection of the pseudoaneurysm and ligation of the injured artery. PMID- 14526649 TI - Achema. PMID- 14526650 TI - [Therapeutic compliance: new frontier in the efficiency of treatment. The example of nitroglycerin transdermal systems]. PMID- 14526651 TI - Effect of Lourdes water on water pH. AB - In samples of normal table water, the level of pH is significantly lower than that of reference samples, when an aliquot of water originating from the springs close to the sanctuary to the Virgin Mary at Lourdes at a final dilution rate of 1:400,000 parts has been added to the sample of water tested. The differences as compared to the reference waters, that is those samples to which the aliquot of Lourdes water were not added, were tested by analysis of the variance and were found to be highly significant. The measurements were conducted on approximately 600 samples. It was demonstrated that the phenomenon observed could not be attributed to chemical species present in the Lourdes water which can justify the lowering of the pH in relation to the dilution rate with which the work was performed. PMID- 14526652 TI - Synthesis and bactericidal properties of ethyl 4-sulfonamido pyrimidine 5 carboxylate derivatives. AB - The paper presents synthesis of ethyl 6-methyl-4-arylamine-4-sulfonamide-2-phenyl 5-carboxypyrimidine derivatives and the results of microbiological tests of new derivatives performed on selected bacterial strains. PMID- 14526653 TI - Comparison of the compliance of pharmacy patients. AB - This issue is an attempt to be compared the compliance, respectively non compliance among pharmacy patients between the years 1998 and 2001. The authors make an attempt to elucidate the factors that influence the non-compliance of the patients in order to improve it and to increase the rate of compliance among them. A questionnaire is applied for assessment the state of compliance and non compliance among the pharmacy patients. The results of the analysis of the received data show that the level of compliance was 69% in 1998 and has decreased to 53% in 2001. The main reasons that influence the non-adherence of the patients are shown. PMID- 14526654 TI - Development of an ophthalmic formulation containing ciprofloxacin-hydroxypropyl-b cyclodextrin complex. AB - Ophthalmic formulations of ciprofloxacin were formulated using inclusion complexes of ciprofloxacin HCl (CFLX HCl) and hydroxypropyl-beta-cyclodextrin (HP beta-CD). The complexes were prepared by the method of freeze drying and evaluated by phase solubility studies, differential scanning calorimetry, powder X-ray diffractiometry, fourier transform infrared spectroscopy and scanning electron microscopy. The ophthalmic solutions formulated using the complexed drug exhibited better stability, biological activity and ocular tolerance in comparison to another ophthalmic solution formulated using uncomplexed drug and also in comparison to a marketed preparation. PMID- 14526655 TI - Determination of cyproterone acetate in plasma samples by high-performance liquid chromatography. AB - A rapid and sensitive high-performance liquid chromatographic method for the determination of cyproterone acetate in human plasma has been developed. The chromatographic separation was performed on an analytical mbondapak C8 column (125 yen 4.6 mm, i.d) with an isocratic mobile phase consisting of methanol-water (62.38 v/v). Using ultra violet detection at 282 nm, the detection limit for cyproterone acetate in plasma was 10 ng/ml. The calibration curve was linear over the concentration range 50-160. 0 ng/ml. Cyproterone was isolated from plasma by liquid-liquid extraction and the recovery was about 90% for plasma. The inter-day and intra-day assay coefficients of variation were found to be less than 10%. PMID- 14526656 TI - Preparation and antibacterial evaluation of some nitrone derivatives with Ge(IV) and Sn(IV) chloride complexes. AB - New complexes of the general formula MCl3.L where L1 = [N-p-chlorophenyl)-alpha (2-xanthatonaphthol)nitrone, L2 = [N-(p-anisole)-alpha-(2 xanthatonaphthol)nitrone]; M = Ge(IV) and Sn(IV) have been synthesized and characterized by elemental analyses, i.r. and molar conductances. The antibacterial activities of these ligands and their complexes have been studied against six species of bacteria in vitro at concentration ranging from 1-10 micrograms/ml. No activity were observed for the both ligands while a remarkable activities were exhibited by some complexes of both ions Ge(IV) and Sn(IV) against S. aureus, B. subtilis and P. mirabilis even more than the effect of antibacterial used in this study. PMID- 14526657 TI - Per-oral extended-release bioadhesive tablet formulation of verapamil HCl. AB - The objective of this study was to formulate a hydrogel-forming bioadhesive drug delivery system for oral administration of verapamil HCl (VP). This system is a non-distintegrating gastro-retentive tablet to allow continuous slow release of VP in the stomach medium where it is more soluble. Different formulate of VP tablets were prepared by compression using various proportions of hydroxypropyl cellulose (HPC) and carbopol 934p (CP). The effect of polymer concentration on the release profile, water uptake and in vitro bioadhesion was studied. Five formulae (F3-F7) exhibited slow release profiles. Formulations F6 (40% HPC and 30% CP) and F7 (40% HPC and 40% CP) had the slowest. They showed 63.6 and 52.2% drug release, respectively, after 12 hours. The kinetic analysis of the release data demonstrated that the bigbest linearity were achieved when data fitted in Higuchi equation rather than zero or first order equations. They had n values close to 0.5 that confirming their Higuchi diffusion. F3 showed the highest swelling index (101.2%), however, the detachment force was intermediate (1.427 N/cm2). Formulae (F4-F7) showed relatively strong in-vitro bioadhesive force. They had detachment forces higher than 1 N/cm2. In general, a delay in drug release and an increase in the in-vitro bioadhesion was seen with the increase in both polymer concentrations. The in vitro data revealed that Formulae F4-F7 served the dual purpose of bioadhesion and sustained release. PMID- 14526658 TI - Synthesis and biological evaluation of some new substituted naphthoquinones. AB - Two novel series of 1,4-naphthoquinone derivatives have been synthesized namely; N-ethoxycarbonyl-2-ethoxycarbonyloxy-3- alkyl-1,4-naphthoquinon-1-substituted phenylhydrazones 3a-f and 2-chlorocetyloxy-3-alkyl-1,4-naphthoquinone-1 substituted phenylhydrazones 4a-d. The antimicrobial activity as well as anticancer activity of these compounds have been evaluated. The acute toxicity of the active compounds was determined. PMID- 14526659 TI - [Incidence and risk indicators of thromboembolism during the acute phase of single chamber ventricular pacing]. AB - BACKGROUND: The incidence of thromboembolism may be higher in single chamber ventricular pacing than in physiological pacing. However, the incidence of thromboembolism during the acute phase of single chamber ventricular pacing (within 14 days) is not known. OBJECTIVES: The incidence and the risk indicators of thromboembolism were investigated during the acute phase of single chamber ventricular pacing. METHODS: Fifty-five consecutive patients (22 males, 33 females, mean age 67 +/- 9 years) who required VVI pacemakers due to brady-tachy sick sinus syndrome (42 patients) or brady atrial fibrillation (13 patients) in the period from April 1975 to December 2000 were retrospectively reviewed. The patients were divided into groups with thromboembolism and without thromboembolism, and the risk indicators for thromboembolism were analyzed. RESULTS: Seven patients (13%) suffered systemic thromboembolism. Three patients had thromboembolic events during temporary ventricular pacing, and four patients had thromboembolic events just after permanent VVI pacemaker implantation. The following risk indicators were identified in the patients with thromboembolism: hyperlipidemia, hypertension, organic heart disease (p < 0.05, respectively), and diabetes mellitus (p < 0.0005). CONCLUSIONS: Patients with brady-tachy sick sinus syndrome and brady atrial fibrillation have a significant risk of thromboembolism during the acute phase of single chamber ventricular pacing. Effective anticoagulation is needed in these patients. PMID- 14526660 TI - [Relationship between duration of arrhythmia and subsequent preventive effect of disopyramide after cardioversion in patients with symptomatic paroxysmal and persistent atrial fibrillation]. AB - OBJECTIVES: The relationship between the duration of arrhythmia and the subsequent long-term efficacy of disopyramide in preventing atrial fibrillation was investigated in patients with symptomatic paroxysmal and persistent atrial fibrillation. METHODS: A total of 60 patients (39 men, 21 women, mean age 65 +/- 11 years) were given disopyramide (300 mg/day) after electrical and pharmacological cardioversion based on American Heart Association Task Force on Practice Guidelines. The patients were divided into two types based on the duration of atrial fibrillation: conversion within 48 hr (group A, n = 35) and more than 48 hr (group B, n = 25) after the episode. Mean follow-up period was 47.1 +/- 28.7 months. RESULTS: Patient characteristics showed no statistically significant difference between groups A and B. The actuarial rates of maintenance of sinus rhythm at 1, 3, 6, 12, 18 and 24 months were 88.6%, 77.1%, 57.1%, 48.6%, 42.9% and 37.1%, respectively, in group A, and 72.0%, 44.0%, 28.0%, 16.0%, 12.0% and 8.0%, respectively, in group B. There was a significant difference in the rate at 24 months between groups A and B (p < 0.05). The periods for maintenance of sinus rhythm in groups A and B were 20.9 +/- 3.9 and 6.7 +/- 2.1 months, respectively, with a significant difference between groups A and B (p < 0.01). CONCLUSIONS: The efficacy of disopyramide in preventing the recurrence of atrial fibrillation varies with the duration of the previous episode. These results demonstrate that it is important to convert to normal sinus rhythm earlier to prevent the recurrence of atrial fibrillation in the long term. PMID- 14526661 TI - [Relationship between insulin resistance and oxidative stress in vivo]. AB - OBJECTIVES: The relationship between oxidative stress in vivo and insulin resistance was examined. METHODS: This study included 87 patients, 46 males and 41 females (mean age 63 +/- 10 years), without coronary artery disease. The homeostasis assessment insulin resistance (HOMA-IR) (fasting blood sugar x fasting immunoreactive insulin/405), a marker for insulin resistance, was measured. The patients were divided into three groups: the noninsulin resistance group (N-IR group) without diabetes mellitus (DM) and with fasting blood glucose level of 126 mg/dl and HOMA-IR < or = 1.73 (n = 44), the insulin resistance group (IR group) without diabetes mellitus and with fasting blood glucose level of 126 mg/dl and HOMA-IR > 1.73 (n = 29), and the DM group (type 2 diabetes mellitus) (n = 14). Urinary 8-iso-prostaglandin F2 alpha (U-8-iso-PGF2 alpha) excretion was measured as a marker of in vivo oxidative stress. RESULTS: There were significantly more obese patients in the IR group than in the N-IR group (62% vs 25%, p = 0.001), and the remnant-like particle cholesterol level was significantly higher in the IR group than in the N-IR group (7.6 +/- 5.2 vs 4.6 +/- 1.5 mg/dl, p < 0.01). Patients in the IR group had a significantly larger number of coronary risk factors. U-8-iso-PGF2 alpha excretion was significantly higher in the IR group and DM groups (201 +/- 86, 191 +/- 136 vs 129 +/- 50 pg/mg. Cr, p < 0.0001, p = 0.01), and there was a significantly positive correlation between the number of coronary risk factors, fasting blood sugar and U-8-iso-PGF2 alpha concentration (correlation coefficient = 0.32, 0.37, p = 0.002, p = 0.0003). Multiple regression analysis showed that remnant-like particle cholesterol, fasting blood sugar and insulin resistance were independent factors for U-8-iso-PGF2 alpha concentration (p < 0.0001, p = 0.0007, p = 0.02). CONCLUSIONS: Insulin resistance, remnant lipoprotein and hyperglyceridemia are deeply involved in oxidative stress in vivo. PMID- 14526662 TI - [Prosthetic valve endocarditis caused by Streptococcus constellatus infection complicated with perivalvular abscess: serial observation by transesophageal echocardiography: a case report]. AB - A 61-year-old man was admitted to an associated hospital because of fever. He had undergone aortic valve and mitral valve replacement 6 years ago, because of rheumatic aortic valve stenosis, and mitral valve stenosis and regurgitation. He had prosthetic valve endocarditis caused by a rare Streptococcus constellatus infection complicated by multiple organ failure and systemic embolism. We considered that surgical treatment was difficult, and continued antibiotic treatment. The inflammatory reaction and fever improved. Prosthetic valve endocarditis is often difficult to identify and treat. Streptococcus constellatus infection is characterized by destruction and formation of abscess. We followed up the patient by transesophageal echocardiography, and observed the course of change of the paravalvular abscess around the aortic valve from echogenic to echolucent. PMID- 14526663 TI - [Infective endocarditis with a huge mitral vegetation related to atopic dermatitis and high serum level of infection-related antiphospholipid antibody: a case report]. AB - A 24-year-old woman with atopic dermatitis was admitted to our hospital with fever. Echocardiography showed a huge vegetation attached to the posterior mitral commissure without mitral valve dysfunction. Blood culture identified methicillin sensitive Staphylococcus aureus. The serum level of antiphospholipid antibody was elevated. A splenic infarction occurred on the second hospital day. Surgery to resect the residual mobile vegetation was performed uneventfully on the 6th hospital day. The postoperative course was uneventful, and the patient was discharged after 4 weeks of antibiotic therapy. Preservation of the mitral valve is rare in the face of virulent Staphylococcus infection and the presence of a huge mobile vegetation. These findings were apparently related to the high serum level of infection-related antiphospholipid antibody and atopic dermatitis. PMID- 14526664 TI - [A 46-year-old woman with faintness and ejection systolic murmur]. PMID- 14526665 TI - Assessment of the Community-Based Surveillance System in Ghana and its role in dracunculiasis eradication. PMID- 14526666 TI - Dracunculiasis eradication: case definition, surveillance and performance indicators. PMID- 14526667 TI - Beyond the suspicious thyroid fine needle aspirate. A review. AB - Fine needle aspiration (FNA) is currently the best diagnostic tool for thyroid nodules. However, the cytologic category of indeterminate or suspicious lesion, which is found in 10-15% of cases, remains a challenge. Since neither clinical presentation nor intraoperative frozen section is often helpful in differentiating these lesions and since surgical procedures for benign and malignant lesions differ, there is a clear need to develop ancillary tests. In this review we identify 12 potential markers of thyroid malignancy that have been examined in thyroid cytologic samples. Although many of these markers hold promise as adjuncts to FNA cytology, multicenter studies have often shown limitations in the predictive value of these assays due to lack of specificity, sensitivity or both. The recent development, however, of tissue microarray techniques to validate promising new markers suggests that improvements in the approach to indeterminate thyroid FNA samples may soon be at hand. This review presents a summary of the issues facing the development of a clinically useful diagnostic test in the differential diagnosis of thyroid nodules. PMID- 14526668 TI - Pancreatic fine needle aspiration. A comparison of computed tomographic and endoscopic ultrasonographic guidance. AB - OBJECTIVE: To compare the sensitivity and specificity of pancreatic fine needle aspiration (FNA) with computed tomography (CT) and endoscopic ultrasound (EUS) guidance. STUDY DESIGN: A 6-year retrospective review included 137 pancreatic FNAs (CT 51, magnetic resonance imaging 2 or EUS 84). Clinical or histologic follow-up was available for 110 patients. RESULTS: Positive (POS), suspicious (SUS), atypical, negative and unsatisfactory (UNSAT) results represented 25.5%, 7.3%, 7.3%, 40.1% and 19.1% of reported diagnoses, respectively. In detecting malignancy, the sensitivities of POS/SUS results were 71% and 42% for CT and EUS, respectively, with negative predictive value (NPV) of 41% and 45%. Also, 35% of UNSAT results were found to have malignancy. However, EUS was used on more difficult lesions, including 82% of small, cystic ones and 75% of small, solid ones. CONCLUSION: Although the apparent performance of percutaneous CT-guided biopsies surpasses that of EUS, this is due to the choice of endoscopy for more difficult lesions. The specificity and negative predictive value of EUS and CT are similar. PMID- 14526669 TI - Clinicopathologic analysis of fine needle aspiration cytology of the thyroid. A review of 1,613 cases and correlation with histopathologic diagnoses. AB - OBJECTIVE: To evaluate the accuracy of fine needle aspiration (FNA) of thyroid lesions at our institution and to ascertain its usefulness in determining the therapeutic approach. STUDY DESIGN: The authors reviewed the results of 1,613 cases of FNA cytology of thyroid nodules performed from 1999 to 2001 at the Department of Pathology, Chonnam National University Hospital. Cytologic diagnoses were compared with histologic diagnoses in 207 cases that included both FNA and thyroid surgery. RESULTS: The sensitivity for the detection of neoplasms (carcinoma and follicular adenoma) was 78.4% and the specificity 98.2%. A false positive diagnosis was made in 1 case (1.8%) and false negative ones in 28 cases (21.5%). The diagnostic accuracy was 84.4%, with a positive predictive value of 99.0% and negative predictive value of 66.3%. The predictive value of a cytologic diagnosis was 100% in papillary carcinoma. CONCLUSION: FNA is a useful test in determining the therapeutic approach of thyroid lesions. PMID- 14526670 TI - Accuracy of cytologic interpretation of pancreatic neoplasms by fine needle aspiration and pancreatic duct brushings. AB - OBJECTIVE: To investigate the accuracy of fine needle aspiration (FNA) specimens and pancreatic duct brushings in the detection of pancreatic lesions and to compare the results with follow-up biopsy and/or surgical interpretation. STUDY DESIGN: We reviewed a total of 57 specimens (37/20), 37 FNA specimens and 20 pancreatic duct brushings, from 45 patients treated at Froedtert Memorial Lutheran Hospital, affiliated with the Medical College of Wisconsin, Milwaukee, over a 4-year period. The FNA and brushing samples were categorized as follows: positive for malignancy (21/3 = 24), suspicious for malignancy (8/7 = 15) and atypical (8/10 = 18). The results were then correlated with the tissue diagnosis. RESULTS: The 24 cytologic samples positive for malignancy included 23 (20/3) pancreatic ductal carcinoma (CA) and 1 (1/0) neuroendocrine CA; in the suspicious category, 11 (6/5) were pancreatic ductal CA; 2 (0/2) mucinous neoplasms and (2/0) neuroendocrine neoplasms; in the atypical category; 2 (2/0) suggestive of mucinous neoplasia, 1 (1/0) suggestive of serous neoplasia and 9 (2/7) favor reactive; and 6 (3/3) without further categorization. Tissue diagnoses were available in 26 cases: 12 (10/2) cases positive for malignancy, 8 (5/3) suspicious for malignancy and 6 (5/1) atypical. The 12 cytologically positive cases confirmed by histology showed 10 ductal CA, 1 neuroendocrine CA and 1 negative. All 8 cases (100%) suspicious for malignancy revealed positive results, including 5 ductal CA, 1 neuroendocrine neoplasm, 1 mucinous cystic neoplasm and 1 lymphoma. Of the 6 atypical lesions, 1 showed ductal CA, 2 mucinous cystic neoplasm and 3 chronic pancreatitis. CONCLUSION: Pancreatic FNA and duct brushings [table: see text] are accurate methods in identifying pancreatic lesions, particularly ductal CA. Accuracy can be improved in the case of mucinous and other lesions with adequate cellularity of the smear and recognizing the limitations of brush samples in the case of mucinous cystic lesions. False negative results may occur in cases of poor representation of malignant cells or poor sampling. PMID- 14526671 TI - Fine needle aspiration cytology diagnosis of lymphoproliferative disease of the breast. AB - OBJECTIVE: To evaluate the role of fine needle aspiration cytology (FNAC) in the diagnosis of lymphoproliferative disease (LPD) of the breast. STUDY DESIGN: Over a period of 20 years (January 1982-December 2001), 13 diagnosed and/or suspected cases of LPD of the breast on FNAC were retrieved and reviewed from the files of the Cytology Laboratory, Department of Pathology, All India Institute of Medical Sciences. For each case, both May-Grunwald-Giemsa- and Papanicolaou-stained smears were reviewed along with hematoxylin and eosin-stained tissue sections and immunohistochemistry, when available. RESULTS: Of the 13 cases, 1 aspirate was from a male breast and the rest were from female. Only 12 cases with documented histology were included in the study. Five of the 12 cases were diagnosed on FNAC as high grade lymphoma, 2 as low grade lymphoma, 2 as poorly differentiated malignant tumor/lymphoma, and 1 each as Hodgkin's lymphoma, acute myeloid leukemia (AML) deposit and immature lymphoid cells, ?leukemia/lymphoma deposit. The tumors manifested mostly as an unilateral mass (10 cases), with 2 cases presenting with bilateral breast lumps. The lymph nodes were involved in 8 cases. Histologically, 2 of the 12 cases were poorly differentiated malignant tumors. Of the remaining 10 cases, 8 were documented as non-Hodgkin's lymphoma and 1 each as Hodgkin's lymphoma, mixed cellularity and AML. CONCLUSION: FNAC is an inexpensive but highly useful diagnostic tool to distinguish between primary lymphoma and carcinoma of the breast. This helps with clinical management in avoiding unnecessary surgical procedures. PMID- 14526672 TI - Cytomorphologic differentiation of Hodgkin's lymphoma and Ki-1+ anaplastic large cell lymphoma in fine needle aspirates. AB - OBJECTIVE: To cytomorphologically differentiate Hodgkin's lymphoma (HL) from Ki 1+ anaplastic large cell lymphoma (ALCL) in fine needle aspirates. STUDY DESIGN: We blindly reviewed 63 fine needle aspiration (FNA) smears from histologically and immunophenotypically proven cases of ALCL (n = 15) and HL (n = 48). The smears were reviewed for the following criteria: (1) estimated percentages of abnormal cells, (2) pattern of the smears (polymorphous vs. dimorphous), and (3) presence or absence of multilobated cells. RESULTS: All cases were phenotyped by immunohistochemistry for CD3, CD15, CD20, CD30 and CD45, with flow cytometric immunophenotpyping in 41 cases. Flow cytometric phenotyping was not successful in any of the cases. The smears were polymorphous in all 15 cases of ALCL and in 1 case of HL (2%). The percentage of abnormal cells ranged from 10% to 90% in cases of ALCL (median, 30%) whereas it ranged from 1% to 25% in HL (median 3%; P = .0003). Three cases of HL showed abnormal cells constituting > or = 20% of the smears. They were all grade 2 disease. Multilobated cells were identified in 14 of the 15 cases of ALCL (93%) and in 3 of the 48 cases of HL (6.25%; P = .0008). CONCLUSION: Our findings indicate that the differentiation of ALCL from HL can be achieved in FNA smears through identification of abnormal cells representing > 30% of the population, a spectrum of abnormal cells and the presence of multilobated nuclei. Rare cases of grade 2 HL may be difficult to differentiate from ALCL. PMID- 14526673 TI - Significance of extensive hyperkeratosis on cervical/vaginal smears. AB - OBJECTIVE: To assess the significance of reporting hyperkeratosis on cervical/vaginal (CV) smears. STUDY DESIGN: Cases diagnosed with extensive hyperkeratosis (E-HK) and without prior or concurrent history of neoplasia, squamous intraepithelial lesion or atypical squamous cells of undetermined significance (ASCUS) were retrieved from our files for the period January 1994 August 2001. E-HK is defined in our practice as patches of anucleated squames with irregular, angulated edges present in at least 5 low-power (10 x eyepiece and 10 x objective) fields on a conventional CV smear. On liquid-based preparations, we use 3 low-power fields. Only cases with a follow-up CV smear and/or cervical biopsy (CB) were selected. RESULTS: Among 328 cases of E-HK, 138 patients met the study selection criteria. Eighty-one cases had negative CV smears and/or CB, 17 (12.3%) patients had persistent E-HK, and a subsequent diagnosis of ASCUS or higher was made in 40 patients (28.9%). Among the 40 cases with subsequent abnormalities, 13 (9.4%) were diagnosed with ASCUS, 24 (17.4%) with HPV or dysplasia, and 3 (2.1%) with malignancy. CONCLUSION: While isolated, anucleate squames may have no clinical importance in patient management, E-HK can be a significant marker of underlying neoplastic disease. This should be kept in mind as one decides how to report CV cytology based on 2001 Bethesda System recommendations. PMID- 14526674 TI - Performance of a new, liquid-based cervical screening technique in the clinical setting of a large French laboratory. AB - OBJECTIVE: To compare the performance of liquid-based cytology with the CYTO screen System (SEROA) with that of conventional smears through a secondary analysis of a large database covering the activity of an independent French laboratory during the period 1998-2002. STUDY DESIGN: The study was performed with a retrospective, comparative, historical design on 2 subgroups of women having been screened by gynecologists who switched from conventional smears to the CYTO-screen System in the period 1998-2002. The first cohort population consisted of women who had at least 4 subsequent screening tests over the period with half conventional and half with the CYTO-screen System. A control group consisted of smears collected by gynecologists who fully maintained activity with a conventional method over the same period. The second group consisted of women who had their first screening test performed over the study period by gynecologists who modified their technique. Specimen adequacy, endocervical cell content and epithelial cell abnormality detection rates were compared between the groups. RESULTS: As compared with the conventional smear, the CYTO-screen System showed a reduction in unsatisfactory reports, especially in the second group of first-screened (0.14% versus 1.3%, P < .0001). The rate of atypical squamous cells of undetermined significance increased significantly after the switch to the CYTO-screen System (2.5% versus 1.3%, P = .004) and in the second group of first-screened women (2.05% versus 1.4%, P = .0014), with higher histologic confirmation in both situations. There was a non-significant increase in the detection rates of low and high grade squamous intraepithelial lesions after the switch to the CYTO-screen System and in the second group of first-screened women. CONCLUSION: The CYTO-screen System gives higher-quality specimens and has a higher detection rate for squamous intraepithelial lesions, but that rate was significant only for atypical squamous cells of undetermined significance. PMID- 14526675 TI - Value of histiocyte detection in Pap smears for predicting endometrial pathology. An institutional experience. AB - OBJECTIVE: To determine the clinical implications of the finding of histiocytes in Pap smears in 1 patient population. STUDY DESIGN: The medical records and Pap smears which the presence of histiocytes was mentioned in the diagnosis between August 1996 and August 2001 were reviewed in conjunction with follow-up surgical findings. The positive predictive value (PPV) for significant endometrial pathology for the isolated finding of histiocytes on Pap smear was determined. RESULTS: Of the 238,225 women screened over a 60-month period, 325 were reported to have histiocytes in their Pap smears. Of them, 238 (73.2%) had subsequent endometrial sampling, hysterectomy or both, and follow-up Pap smears. Two hundred seven smears (87%) failed to disclose endometrial pathology. Thirty-one cases (13%) resulted in significant histopathologic findings, including 12 uterine malignancies, 8 endocervical polyps, 7 endometrial polyps, 2 submucosal leiomyomata, 1 simple hyperplasia without atypia and 1 case of tamoxifen-related changes. Upon review of the clinical records, 58% (18/31) of those patients had other significant clinical and/or cytologic findings. Five of the 18 patients (27.8%) had associated postmenopausal bleeding, 11 had additional abnormal Pap smear findings (atypical glandular cells, 6/18, or 33.3%; endometrial cells, 5/18, or 27.8%), and another 2 had both postmenopausal bleeding and atypical glandular cells (2/18, or 11.1%). The PPV for significant uterine pathology for women with the isolated finding of histiocytes on a Pap smear was 5.5% and 60% with additional clinical and/or Pap smear findings. The PPV for endometrial cancer was 1.3% in women with the isolated finding of histiocytes on a Pap smear but 20% for women with histiocytes and additional clinical/or Pap smear findings. CONCLUSION: Based on the findings of this study and recently published data, we conclude that the isolated finding of increased histiocytes in the absence of postmenopausal bleeding, endometrial cells or atypical glandular cells on a Pap smear is a poor indicator of uterine disease. PMID- 14526677 TI - Detecting mycobacteria in Romanowsky-stained cytologic smears. A case report. AB - BACKGROUND: Cytologic features of mycobacterial infections are granulomatous inflammation with or without caseous necrosis and the demonstration of acid-fast bacilli with special stains. However, immuno-compromised patients might not mount the expected response. CASE: A routinely used Romanowsky (Leishman) stain was used for the presumptive diagnosis of mycobacterial infection in a 30-year-old man with AIDS. The mycobacteria were identified as inclusions, described as "negative images," in the cytoplasm of macrophages in smears of bone marrow aspirate. They were then confirmed to be acid-fast bacilli with Ziehl-Neelsen stain. CONCLUSION: Negative images of mycobacteria may be seen in Romanowsky stained cytologic smears from patients with immunodeficiency. This is a rapid and cost-effective way of detecting the mycobacteria before more specific results are available. Such a search should be undertaken routinely in all patients suspected to have such infections. PMID- 14526676 TI - Imprint cytology of sentinel lymph nodes in breast cancer. Experience with rapid, intraoperative diagnosis and primary screening by cytotechnologists. AB - OBJECTIVE: To evaluate the intraoperative imprint diagnoses of smears from sentinel lymph nodes that had been primary screened by cytotechnologists and to assess the most important causes of false negative (FN) imprint diagnoses. STUDY DESIGN: Material consisted of 429 imprints from sentinel lymph nodes in 211 breast cancer patients that were sent for frozen section examination over 13 months. RESULTS: The mean number of imprints/lymph nodes per patient was 2.02. The mean screening time per imprint was 3.6 minutes. Sixty-six sentinel nodes (16%) from 51 women (24%) were metastatic. Imprints and/or frozen sections were positive in 54 nodes (82%). Imprints were positive in 38 nodes, representing 70% of intraoperative positive nodes and 58% of the total number of positive nodes. Twenty-six of 28 (93%) FN imprints were due to suboptimal sampling. Four of 9 FN macrometastases did not contain diagnostic or suspicious cells/cell groups even on rescreening, whereas a few, and then only 1 diagnostic group were identified in 2/9. There were no false positives. CONCLUSION: Primary screening by experienced cytotechnologists is both rapid and reliable and enabled the diagnosing pathologist to concentrate on the frozen section. The major cause of false negative imprints is sampling, even in macrometastases. PMID- 14526678 TI - Coccidioidomycosis osteomyelitis masquerading as a bone tumor. A report of 2 cases. AB - BACKGROUND: Coccidioidomycosis involving the lung is common in endemic areas. However, it is unusual for it to present as a solitary bone lesion. CASES: Two cases of Coccidioides immitis osteomyelitis clinically and radiographically mimicked a primary bone tumor. Fine needle aspiration (FNA) biopsy of these lytic bone lesions yielded diagnostic material. Aspirated material in 1 case showed a suppurative, granulomatous inflammation and scattered spherules with refractile walls, some containing endospores consistent with coccidioidomycosis. Smears in the other case showed acute inflammation and necrosis, and rare spherules were identified with the Gomori-mathenamine-silver stain. C immitis infection was confirmed by culture in both cases. CONCLUSION: Although rare, coccidioidomycosis can involve the bone and mimic a primary bone tumor. FNA biopsy is helpful in differentiating between inflammatory and neoplastic processes involving bone by acquiring material for cytologic studies and cultures. PMID- 14526679 TI - Ultrasound-guided fine needle aspiration cytodiagnosis of leiomyosarcoma metastatic to the breast. A case report. AB - BACKGROUND: Leiomyosarcomas are characterized by slow growth and late metastasis and most often involve the lung, liver and subcutaneous tissue. Metastasis to the breast is rare, with only four cases published in the English-language literature to date. Because of its rarity and the prolonged time interval between the diagnosis of the primary tumor and involvement of distant sites, accurate diagnosis of these tumors can be challenging. Evaluation of the cytomorphologic features of the tumor, ancillary immunocytochemical staining and detailed medical history of the patient are essential to making a correct diagnosis. CASE: A case of leiomyosarcoma metastatic to the breast occurred in a 60-year-old woman whose primary uterine leiomyosarcoma had been resected 10 years before the diagnosis of the metastasis. Fine needle aspiration performed with ultrasound guidance yielded a moderately cellular specimen composed of minimally pleomorphic spindle cells with smooth, blunt-ended nuclei arranged in an interlacing pattern. Immunocytochemical staining for muscle-specific actin confirmed the smooth muscle origin of the neoplasm. CONCLUSION: The results of cytomorphologic analysis and immunocytochemical staining performed after fine needle aspiration and the patient's history of primary uterine leiomyosarcoma allowed us to correctly diagnose metastasis to the breast and avoided inappropriate management of the metastasis as a primary tumor. PMID- 14526680 TI - Plasmacytoma with involvement of the urinary bladder. Report of a case diagnosed by urine cytology. AB - BACKGROUND: Plasmacytoma of the bladder is a rare but important entity. We report a case of plasmacytoma of the bladder that was diagnosed by urinary cytology. CASE: A 71-year-old male with a history of multiple myeloma presented in renal failure. Renal ultrasound revealed right-sided, moderate hydronephrosis with a 4 x 4-cm, posterolateral, obstructing mass. Magnetic resonance imaging demonstrated a bladder mass involving the bladder base, right lateral wall and dome with extension into the perivesical tissues on the right. The mass showed a moderate degree of enhancement following intravenous gadolinium administration. Urine cytology was performed to evaluate for bladder carcinoma or other malignancies besides plasmacytoma. The specimen was signed out as multiple myeloma of the bladder. Cystoscopy and biopsy were subsequently performed on the bladder mass. The diagnosis of plasmocytoma was made, confirming the urine cytology diagnosis. CONCLUSION: Urinary cytology can be a diagnostic tool for plasmocytoma involving the bladder. PMID- 14526681 TI - Adenoid cystic carcinoma of the external auditory canal. A case report with diagnosis by fine needle aspiration. AB - BACKGROUND: Although the cytologic findings of adenoid cystic carcinoma have been reported frequently, fine needle aspiration diagnosis of adenoid cystic carcinoma of the external auditory canal has not been reported before in the English language literature. CASE: A 70-year-old female presented with a mass occluding the external auditory canal. Fine needle aspiration cytology was performed. The smears showed hyaline globules surrounded by tumor cells. The tumor cells had uniform, round, hyperchromatic nuclei, visible nucleoli and little cytoplasm. Based on these cytomorphologic features, diagnosis of adenoid cystic carcinoma was rendered and then confirmed by histopathologic examination. CONCLUSION: This is the first cytologic report of adenoid cystic carcinoma of the external auditory canal, a rare site for this tumor. PMID- 14526682 TI - Acantholytic cells exfoliated from pemphigus vulgaris of the uterine cervix. A case report. AB - BACKGROUND: Pemphigus vulgaris of the uterine cervix is rare and almost always associated with cutaneous or mucosal lesions elsewhere on the body. Without a history of pemphigus, acantholytic cells in cervical smears may be misdiagnosed as malignant ones. CASE: A 52-year-old female presented with a vaginal discharge, and a routine cervical smear was collected for cytology. The smear displayed atypical cells lying singly and in loose clusters, having vesicular nuclei, a thin nuclear membrane, prominent nucleoli and well-defined cytoplasmic margins. These were labeled atypical glandular cells of undetermined significance, and colposcopic examination and biopsy were advised. On colposcopy vesicular lesions and erosions were noticed on the cervix. The biopsy revealed typical intact as well as denuded suprabasal bullae of pemphigus vulgaris. On reevaluation of the cytologic smear, the cytomorphologic features correlated well with the acantholytic cells of pemphigus. Thorough reexamination of the patient revealed 2 tiny vesicles on the oral mucosa that, on biopsy, confirmed the diagnosis of pemphigus vulgaris. CONCLUSION: Cytopathologists should be aware of the typical cytomorphologic features of pemphigus vulgaris and, in an appropriate clinical setting, should be able to diagnose or at least suspect this entity in even rare sites like the cervix. A false positive diagnosis of malignancy can be avoided if the cytologic findings are judiciously correlated with the history and with clinical and colposcopic examinations. PMID- 14526683 TI - alpha-Fetoprotein-producing liver carcinomas of primary extrahepatic origin. AB - BACKGROUND: alpha-Fetoprotein (AFP)-producing carcinomas, hepatoid or otherwise, are increasingly being recognized at extrahepatic sites. Some of them not only mimic hepatocellular carcinomas (HCCs) in having a proclivity for vascular permeation and distant metastases but also exhibit identical morphology and immunoreactivity for alpha-1-antitrypsin and HepPar1. beta-Human chorionic gonadotropin (hCG) is also detected. This would create diagnostic problems in hepatic fine needle aspiration biopsies (FNABs) from patients with elevated serum AFP. Apart from HCC, its variants and germ cell tumors, one must consider metastatic AFP-producing carcinomas. CASES: A man with gastric adenocarcinoma had a liver mass. Hepatic FNAB revealed an AFP-producing adenocarcinoma. The gastric tumor was positive for AFP, polyclonal carcinoembryonic antigen, HepPar1, CK19, hCG and synaptophysin. A woman with endocervical adenocarcinoma had multiple liver nodules. FNAB revealed an AFP-producing, undifferentiated carcinoma. The cervix showed a large cell neuroendocrine carcinoma coexisting with an adenocarcinoma in situ. The large cells were positive for synaptophysin, AFP, hCG and AE1/3. The glands showed diffuse HepPar1 and focal synaptophysin expression. CONCLUSION: A wide histologic spectrum of extrahepatic carcinomas can produce AFP and other peptide hormones. The true AFP status probably is not recognized at the first presentation. Such carcinomas, whether hepatoid or not, behave aggressively. Their recognition at the initial presentation is crucial to early and appropriate therapy. These entities add a new dimension to the challenges of FNAB diagnosis. PMID- 14526685 TI - Cellularity of liquid-based, thin-layer cervical cytology. PMID- 14526684 TI - Anaplastic large cell lymphoma presenting as a pleural effusion and mimicking primary effusion lymphoma. A report of 2 cases. AB - BACKGROUND: Systemic anaplastic large cell lymphoma (ALCL) is predominantly a nodal disease, but extranodal involvement can occur during the disease course or as the primary presentation. We report two rare cases of ALCL presenting with a pleural effusion, mimicking primary effusion lymphoma (PEL). CASES: Two patients, a 47-year-old woman and an 81-year-old man, presented with a pleural effusion for investigation. The pleural fluid contained abundant, large, lymphoid cells with marked nuclear atypia. These neoplastic cells strongly expressed CD30 and EMA and showed a T-cell phenotype (CD3+CD45RO+ for case 1 and CD4+ for case 2). Case 1, in addition, showed ALK1 expression. The tumor cells in both cases were negative for human herpes virus type 8 (HHV8) and Epstein-Barr virus (EBV). ALCL shows overlapping cytologic features with PEL, but the T-cell phenotype, ALK1 expression in case 1, lack of association with HHV8 and EBV, HIV seronegativity and subsequent discovery of nodal disease in case 2 were all in favor of ALCL over PEL. CONCLUSION: In rare cases a pleural effusion is the presenting feature of ALCL, and distinction from PEL depends on correlation with clinical findings, detailed immunophenotyping and study of the status of HHV8 and EBV. PMID- 14526686 TI - Endometriosis in the differential diagnosis of abdominal wall masses. PMID- 14526687 TI - Cytologic imprints in intraoperative consultations for gastric heterotopic pancreas. PMID- 14526688 TI - Granulocytic sarcoma presenting as a breast mass. PMID- 14526689 TI - Population-based Pap screening in Cameroon. PMID- 14526690 TI - "Daisy" cells in peritoneal washings. PMID- 14526691 TI - Investigation of an outbreak of Legionnaires' disease in a hospital under construction: Ontario, September-October 2002. PMID- 14526692 TI - Prevention of pertussis in adolescents and adults. PMID- 14526693 TI - Supplementary statement on conjugate meningococcal vaccines. PMID- 14526694 TI - Permanent junctional reciprocating tachycardia: an incessant tachycardia in children. AB - Permanent junctional reciprocating tachycardia is an infrequent form of reentrant supraventricular tachycardia but is the commonest incessant tachycardia in childhood. The persistent nature of the tachycardia can lead to heart failure secondary to left ventricular dysfunction and the arrhythmia is often refractory to drug control. STUDY OBJECTIVE: To review the main clinical features and therapeutic options currently available for this arrhythmia. DESIGN: Retrospective study. PATIENTS: A group of 5 children with a diagnosis of permanent junctional reciprocating tachycardia. RESULTS: Age at presentation ranged between 14 days and 12 years. Three were asymptomatic. The first echocardiogram displayed left ventricular dilation and decreased shortening fraction in one child. Pharmacological therapy was initially successful in all. During follow-up (0.2-4.5 years) the arrhythmia became resistant to medication in one case and radiofrequency ablation was carried out. CONCLUSIONS: Permanent junctional reciprocating tachycardia has a wide range of clinical presentations. Pharmacological therapy is recommended, but radiofrequency ablation is the definitive treatment and only depends on patient age. PMID- 14526695 TI - Electrical disconnection versus anatomical isolation of the pulmonary veins: two sides of the same treatment for atrial fibrillation? AB - OBJECTIVE: There are currently two techniques for percutaneous electrical isolation of the pulmonary veins (PV): anatomical isolation and electrical disconnection. The aim of the present study was to assess the continuity and circumferential extension of the radiofrequency applications necessary for PV electrical disconnection in order to evaluate the differences between this technique and anatomical isolation. METHODS: We studied 9 patients with paroxysmal atrial fibrillation who underwent PV electrical disconnection. The electrophysiologic study was performed with a decapolar circular catheter (Lasso) introduced in the PV ostia and a mapping and ablation catheter (CARTO). Ablation was performed following the activation sequence of PV potentials. Using the CARTO system we evaluated the number of applications, their distribution in quadrants, and the maximum distance between contiguous and opposite RF applications. We assessed the number of veins with circumferential applications. The number of applications and maximum distance between contiguous applications were compared to the maximum distance between opposite applications. RESULT: A total of 26 PV were isolated, including 9 right superior, 5 right inferior, 6 left superior and 6 left inferior. On average more than 10 applications were necessary, with a heterogeneous distribution. In 80.8% of the PVs the maximum distance between contiguous application was less than 1 cm and maximum distance between opposite application were highest in the right superior PV and lowest in the right inferior PV. The applications were circumferential in 80.8% of the disconnections. The number of RF applications was significantly higher in PVs with greater distances between opposite applications (correlation coefficient 0.51; p=0.008). No relation was found between maximum distances in contiguous and opposite applications (correlation coefficient r=0.13; p=NS). CONCLUSION: Electrical disconnection was achieved in the great majority of cases with circumferential applications, similarly to anatomical isolation. However, the heterogeneous distribution of the applications implies the existence of areas that need a greater number of applications and that can only be identified during electrophysiologic study. Therefore, rather than two aspects of the same treatment for atrial fibrillation, electrical disconnection and anatomical isolation complement each other in the achievement of effective PV isolation. PMID- 14526696 TI - Contrast echocardiography in segmental analysis and intraventricular gradient quantification in hypertrophic cardiomyopathy. AB - INTRODUCTION: Hypertrophic cardiomyopathy (HCM) is a hereditary disorder characterized by ventricular hypertrophy, diastolic dysfunction and hyperdynamic left ventricular systolic function. This excessive contraction is sometimes associated with significant intraventricular pressure gradients. These gradients are dynamic and therefore vary at different times. Echocardiography can identify and quantify the functional and morphologic changes characteristic of the disease. Ultrasound contrast agents (UCAs) are indicated in patients with poor transthoracic image quality, enabling better visualization of the endocardial border. These agents also strengthen the Doppler signal, which enables better quantification of the transvalvular and intraventricular gradients. In HCM, definition of the endocardial/blood interface and visualization of the myocardial structure, as well as quantification of intraventricular gradients, are fundamental to the study of the pathology. OBJECTIVES: The objective of this study was to evaluate the clinical utility of new UCAs in morphologic study (segmental analysis) and quantification of maximum intraventricular gradients (IVG Max) in HCM, as well as the feasibility and interest of determining mean gradients (IVG Med) in HCM. METHODS: Thirty-four patients with clinical and echocardiographic diagnosis of HCM were studied. Baseline IVG was considered significant when over 30 mmHg. Left ventricular morphology and IVG quantification were assessed before (study A) and after (study B) UCA injection. Maximum (Max) and mean (Med) values of delta IVG were calculated. Endocardial border definition of ventricular segments was analyzed in studies A and B and the percentage of ventricular segments that were completely visualized throughout the cardiac cycle was established. RESULTS: The mean values of delta IVG Max for studies A and B were 51 +/- 31 mmHg and 61 +/- 32 mmHg, p = NS. The mean values of delta IVG Med were 26 +/- 16 mmHg in study A and 31 +/- 17 mmHg in study B, p = NS. The correlation between delta IVG Max and Med in study A was r2 = 0.74, p < 0.01, while in study B it rose to a value of r2 = 0.82, p < 0.01. Segmental analysis: In all segments studied the visualization percentage was higher after UCA injection, with a statistically significant difference in all lateral and anterior wall segments. DISCUSSION AND CONCLUSIONS: HCM is usually evaluated in a non-invasive way by echocardiography. There are no references to systematic use of UCAs in HCM patients. The value of determining the maximum gradient in HCM is generally accepted, but the importance of the mean gradient is not known. In this work, UCAs improved the Doppler signal without distorting values. In HCM, values measured using UCAs have a better correlation, with a smaller discrepancy between Max and Med gradients. This study suggests that mean gradient determination enables better characterization of the dynamic variability of the gradients because there is a correlation between Max and Med gradients. The real importance of mean gradients is not yet established, so further studies are necessary. In conclusion. UCAs are very useful in morphological assessment. The interest of UCAs in determining intraventricular gradients and the value of mean gradients in HCM are not clearly demonstrated in this study. PMID- 14526697 TI - Congenital absence of left pericardium. AB - The authors present a clinical case of right chamber dilatation in a 34-year-old patient with no symptoms, investigation of which led to a diagnosis of congenital absence of left pericardium, established by magnetic resonance imaging (MRI). They also present a review on absence of the pericardium, and discuss associated congenital abnormalities, the clinical presentations and findings in diagnostic tests, with particular emphasis on MRI. PMID- 14526698 TI - Chagas' disease and the involvement of the autonomic nervous system. AB - Chagas' disease is a major endemic disease in Latin America and a great cause for concern due to its high incidence: it afflicts 16 to 18 million individuals and places over 90 million people at risk of infection. At present, five mechanisms can be proposed to explain the pathogenesis of chronic Chagas cardiopathy: 1. direct lesion of the tissue by Trypanosoma cruzi; 2. dysfunction of the autonomic nervous system (neurogenic concept); 3. microvascular disease; 4. immunologic reaction; 5. alterations in the extracellular matrix. The neurogenic concept is the most attractive explanation for the pathogenesis of chronic Chagas cardiopathy through the involvement of the autonomic nervous system, an issue that has been prominent ever since Chagas first initiated research in the field. Koberle, in his pioneering studies on the role of the autonomic nervous system in Chagas patients in the 1950s, adopted the technique of neuron counts, whereby he registered a reduction in parasympathetic nerve cells, and thus considered Chagas cardiopathy a "parasympathetic reduction" with predominance of the sympathetic. In the 1960s, systematic studies on autonomic function, organized by Professor Dalmo Amorim, were initiated in the School of Medicine in Ribeirao Preto. Several aspects of cardiac autonomic control were later described independently by teams in Brazil (Ribeirao Preto and Brasilia), Argentina (Cordoba) and Venezuela (Merida). In general, the studies performed in Ribeirao Preto by Amorim and Marin Neto and in Brasilia by Junqueira Jr. reflected the functional involvement of the parasympathetic system, while the studies performed in Cordoba were linked with the view of cardiovascular sympathetic dysfunction. In Brazil, the involvement of the sympathetic system, with relation to the functional aspect of sympathetic denervation, is well characterized by Marin Neto through the assessment of heart rate using the tilt test in both Chagas and control groups. Further evidence of autonomic nervous system dysfunction in Chagas' disease as a factor modulating complex ventricular arrhythmias was demonstrated by Pedrosa (RJ), who reported on a specific group of chronic Chagas patients with complex ventricular arrhythmias and dilated cardiopathy. In this study, when serum from chronic Chagas patients showing neither complex ventricular arrhythmias nor ventricular dilation was inoculated in isolated rabbit hearts, it produced no harmful effect in the conduction system, in contrast to what was observed in the conduction system of rabbits inoculated with serum from the Chagas patients group with complex ventricular arrhythmias and ventricular dilatation. These facts confirm Carlos Chagas as the pioneer in postulating involvement of the autonomic nervous system in Chagas' disease, and provide an important opportunity to understand ventricular involvement in chronic Chagas cardiopathy. PMID- 14526699 TI - Incidence and prevalence of hypercholesterolemia in Portugal: a systematic review. Part III. AB - INTRODUCTION: Over 50% of the mortality and disability caused by ischemic heart disease and stroke could be avoided by controlling individual risk factors (hypertension, hypercholesterolemia, smoking and obesity). The ignorance of the extent and impact of hypercholesterolemia (HC) in Portugal prompted us to undertake this systematic review of the prevalence and incidence of hyperlipidemia in Portugal, based exclusively on work published nationally. METHODS: We included every study published in the country that could provide data on the prevalence or incidence of hypercholesterolemia. The search strategy included an electronic search of national and international biomedical databases (Medline, Index Revistas Medicas Portuguesas, the Portuguese Directorate-General for Health and the Portuguese National School of Public Health), screening of reference lists from the individual studies, and personal contacts with institutions, groups and authors (such as the Portuguese Foundation of Cardiology). The data--recorded in electronic format--was collected independently by two of the authors (JC and MB), with consensus achieved with a third (AVC) when there were differences in the study coding. RESULTS: We could only identify one paper on the incidence of HC in Portugal, which gave an incidence of 559 new cases per 100,000 inhabitants, increasing with age up to 54 years for men and 64 years for women. There was a higher incidence in men than in women up to the age of 54, but at more advanced ages this relationship was reversed. Prevalence studies on HC included 53,445 individuals overall, with sample size lower than 1000 in most of the individual studies. The most frequently used criterion for HC was > 200 mg/dL, with the mean level being higher in most studies. We calculated the mean prevalence for several cut-off points (> or = 190 mg/dL: 63.8%; > or = 200 mg/dL: 56.7%; > or = 240 mg/dL; 31.7%; > or 250 mg/d; 21%). Due to the heterogeneity of the data, these results are to be interpreted with caution, even though they are consistent across individual studies. The prevalence of hypertriglyceridemia (HTG) was highly variable across studies. Most of them used a baseline criterion of 150 mg/dL (11-26%) or > or = 200 mg/dL (8.9-33.9%). CONCLUSIONS: Despite the obvious heterogeneity and a certain paucity of studies, we found consistent results concerning the prevalence of hypercholesterolemia in Portugal, and we can state that this constitutes a serious health problem in our country. The prevalence of hypercholesterolemia was higher than that of arterial hypertension in our population. The results of this study should serve as a basis for a commitment to action from the health authorities and health professionals as well as patients. PMID- 14526700 TI - Obstruction to the pulmonary venous flow in an infant. PMID- 14526701 TI - Unstable angina--when to proceed to percutaneous coronary intervention. PMID- 14526702 TI - Percutaneous coronary intervention in cardiogenic shock. PMID- 14526703 TI - Percutaneous coronary intervention in acute myocardial infarction. PMID- 14526704 TI - Update in percutaneous coronary interventions. PMID- 14526705 TI - Left-handedness: in support of the ten-percenters. PMID- 14526706 TI - The spirit of sacred community. PMID- 14526707 TI - Mind-body medicine for stress management and opportunities at NCCAM. PMID- 14526708 TI - An approach to the nutritional management of autism. PMID- 14526709 TI - Acupuncture: on what should its evidence base be based? PMID- 14526710 TI - Chinese medicine and cancer care. PMID- 14526711 TI - Developing methods for acupuncture research: rationale for and design of a pilot study evaluating the efficacy of acupuncture for chronic low back pain. AB - Rigorously evaluating acupuncture and other complementary and alternative medicine (CAM) therapies presents researches with many challenges. The failure to satisfactorily address these challenges has resulted in poorly designed studies, which yield findings that are difficult to interpret. Despite the publication of more than 10 randomized trials evaluating acupuncture as a treatment for chronic low back pain, the efficacy and effectiveness of acupuncture for this common problem remain unclear. We discuss the rationale for and design of a five-arm randomized controlled pilot clinical trial that addresses the major methodological shortcomings of previous studies (e.g., poorly justified treatment and control groups and lack of masking) and that lays the groundwork for a full scale trial evaluating acupuncture as a treatment for chronic low back pain. Although development and pilot testing of these design features required substantial time and resources, we believe that this investment in rigorous groundwork is essential to ensure that full-scale trials evaluating CAM treatments produce valid and interpretable results. PMID- 14526712 TI - Effects of total glycosides from Baishouwu on human breast and prostate cancer cell proliferation. AB - Baishouwu (Cynanchum Auriculalum Royle ex Wight), a traditional Chinese herbal medicine, is used as a traditional anti-aging remedy in China. Previous studies suggested Baishouwu has multiple pharmacological actions. In this study, the total glycosides extracted from Baishouwu (CA/TG) were evaluated for its potential anti-tumor activity and mechanism on several human breast and prostate cancer cell lines. In [3H] thymidine incorporation assay. CA/TG inhibited the proliferation of cancer cells in a dose-dependent fashion. It also displayed anti androgen activity on androgen sensitive cell line assessed by reporter gene assay. The morphological examination of CA/TG treated cells revealed overall shrinkage of nucleus, chromatin condensation and small nuclear fragments by DAPI staining, the characteristic pattern of DNA ladders were observed in cells exposed to CA/TG, indicating the involvement of apoptosis in its action. These results suggested that the CA/TG has preferred cytotoxicity against the cancer cells, and the action was due to its ability of preventing cell proliferation and inducing apoptosis. PMID- 14526713 TI - Efficacy of extract of Pelargonium sidoides in children with acute non-group A beta-hemolytic streptococcus tonsillopharyngitis: a randomized, double-blind, placebo-controlled trial. AB - BACKGROUND: Clinical trial data suggest that antibiotics are not indicated for the treatment of acute non-group A beta hemolytic strep (non-GABHS) tonsillopharyngitis. Nevertheless patients are symptomatic and effective alternatives for its treatment are needed that have been evaluated in clinical trials. OBJECTIVE: To confirm that treatment with an extract of Pelargonium sidoides (EPs 7630) is superior to placebo for the treatment of non-GABHS tonsillopharyngitis in children. DESIGN: Randomized, double-blind, placebo controlled trial. SETTING: Six study sites in 4 pediatric and ENT primary care outpatient clinics. PATIENTS: One hundred forty-three children aged 6-10 years with non-GABHS tonsillopharyngitis present < or = 48 h, a negative rapid strep screen, a Tonsillopharyngitis Severity Score (TSS) > or = 8 points, and informed consent. INTERVENTION: EPs 7630 or placebo (20 drops tid) for 6 days. MEASUREMENT: The primary outcome criterion was the decrease of the TSS from baseline (day 0) to day 4. RESULTS: The decrease of the TSS from baseline (day 0) to day 4 was 7.1 +/- 2.1 points under EPs 7630 (n = 73), and 2.5 +/- 3.6 points under placebo (n = 70). The covariate adjusted decrease was 7.0 +/- 2.4 points under EPs 7630, and 2.9 +/- 2.4 points under placebo. The 95% RCI for the difference between the groups was [2.7; 4.9] demonstrating a significant difference in efficacy of EPs 7630 compared to placebo (P < 0.0001). Adverse events (AEs) occurred in 15/143 patients (EPs 7630: 4/73 patient, placebo: 44/70) and were not related to the investigational medication. CONCLUSIONS: EPs 7630 was superior compared to placebo for the treatment of acute non-GABHS tonsillopharyngitis in children. Treatment with EPs 7630 reduced the severity of symptoms and shortened the duration of illness by at least 2 days. PMID- 14526714 TI - Treatment for seasonal allergic rhinitis by Chinese herbal medicine: a randomized placebo controlled trial. AB - CONTEXT: Chinese herbal medicine (CHM) is widely used to treat seasonal allergic rhinitis (SAR), however, evidence of efficacy is lacking. OBJECTIVE: To evaluate the efficacy of a Chinese herbal formulation for the treatment of SAR. DESIGN: Randomized, double blind, placebo controlled trial. SETTING: RMIT Chinese Medicine Clinic. PATIENTS: 55 patients with seasonal allergic rhinitis (active 28, placebo 27). INTERVENTIONS: CHM extract capsule (containing 18 herbs) or placebo, given daily for 8 weeks. MAIN OUTCOME MEASURES: The primary measure of efficacy were changes in severity of nasal and non-nasal symptoms using a Five Point Scale (FPS) measured by both patients and the practitioner. The secondary measure was the change in score for the domains measured in the Rhinoconjunctivitis and Rhinitis Quality of Life Questionnaire (RQLQ) assessed by patients. RESULTS: Forty-nine patients completed the study (active 24, placebo 25). After eight weeks, the severity of nasal symptoms and non-nasal symptoms were significantly less in the active treatment group than in the control group, both for measurements made by patients and those by the practitioner. Comparison of active and placebo treatment groups RQLQ scores also indicated significant beneficial effects of treatment (end point Section 1: P < 0.05; Section 2: P < 0.01). Intention-to-treat analyses of categorical items showed moderate to marked improvement rates were 60.7% and 29.6% for active and placebo respectively. Eleven patients reported mild adverse events including 1 withdrawn from the trial. CONCLUSIONS: This CHM formulation appears to offer symptomatic relief and improvement of quality of life for some patients with seasonal allergic rhinitis. PMID- 14526716 TI - Charlotte Eliopoulos, RN, MPH, PhD, ND. Holistic nursing and spiritual healing. Interview by Bonnie Horrigan. PMID- 14526715 TI - Visit time as a framework for reimbursement: time spent with chiropractors and acupuncturists. AB - CONTEXT: Visit length is an important component of physician payment systems. As use and insurance coverage of complementary and alternative medicine (CAM) increases, equitable reimbursement strategies for CAM providers will be needed. Little information is available about the time CAM providers spend with patients. OBJECTIVE: To describe the length of visit and determinants of visit time to chiropractors and acupuncturists. DESIGN: Survey of CAM practitioners that collected information on providers (e.g., socio-demographics, practice patterns, training) and professional visit encounters (e.g., patient demographics, payment source, problem acuity, visit length, procedures performed). SUBJECTS: Random sample of 130 licensed chiropractors and 133 licensed acupuncturists surveyed in 1998-1999, each from one Western and one Northeastern US state. MEASURES: Patient, provider, and visit factors associated with visit duration were analyzed using weighted linear regression. RESULTS: Data were available on 2550 chiropractic and 2561 acupuncture visits. Mean chiropractic visit length was 21.5 minutes (SE = 0.8). Mean acupuncture visit length was 56.6 minutes (SE = 0.7) In both professions, new patient visits took longer, while visits for wellness or chronic problems were shorter. Preventive counseling by the chiropractor increased visit time, as did use of manual (vs. instrument) spinal manipulation, soft tissue techniques and physiotherapeutics. Acupuncture visits were shorter when specialized needling techniques (e.g. auricular or scalp acupuncture) were used, and longer with other adjunctive Asian therapies (e.g., cupping or magnets). Self-paid acupuncture visits were longer, as were visits with acupuncturists who had less practice experience or fewer years of training. CONCLUSIONS: For both chiropractic and acupuncture, certain visit factors, provider characteristics, and procedures increase visit length, many of which parallel those observed in conventional medical settings. Thus, a similar time based payment scheme may be a reasonable starting point for developing methods for reimbursing CAM providers. PMID- 14526717 TI - Cochrane for CAM providers: evidence for action. PMID- 14526718 TI - The Conemaugh Health System Integrative Health Center. PMID- 14526719 TI - Effect of herbal therapy on chronic herpes virus infections. PMID- 14526720 TI - Amici curiae and the public interest in medical law cases. PMID- 14526721 TI - Access to infertility treatment: lessons from Patrick's story. PMID- 14526722 TI - Falls and fall-related injuries: far-reaching implications. PMID- 14526723 TI - PD v Harvey and Chen [2003] NSWSC 487. PMID- 14526724 TI - Quintavalle v human fertilisation and embryology authority [2003] EWCA Civ 667. PMID- 14526725 TI - Polygraph testing for deception in Australia: effective aid to crime investigation and adjudication? AB - Polygraph testing--or the monitoring and analysing of selected physiological measures of an individual who is being interviewed, for the purpose of detecting deception--is controversial in Australia. Considerable mythology surrounds this method of detecting deception. Embedded in popular perceptions of crime fighting and utilised in high-profile criminal cases in Western Australia, Victoria and Queensland, polygraph testing is also explicitly prohibited from being used in crime investigation in New South Wales. Unlike in the United States, polygraph testing has not hitherto routinely been used by government departments and authorities as a preemployment screening tool, but is increasingly being offered in the private sector in Australia. This article examines the current scope of polygraph testing in Australia, describes different approaches to testing, briefly reviews recent information relating to validity and evaluates State legislation prohibiting the use of polygraph testing for specified purposes in New South Wales. Consequent to the continuing controversy regarding the accuracy of polygraph testing in detecting deception (and, conversely, truth-telling), it is argued that the emerging use of polygraph testing is problematic and common law principles rendering the results of such testing inadmissible in court do not constitute sufficient safeguard against inappropriate and intrusive testing. Future research should identify specific polygraph testing techniques and consider the most socially beneficial way of regulating this emerging area of practice. PMID- 14526726 TI - Perceptions of responsibility for clinical risk management: evidence from orthopaedics practitioners, practice managers and patients in an Australian capital city. AB - The article describes a study of the perceptions of three groups--patients, orthopaedic surgeons and the surgeons' practice managers--concerning three types of legal risk associated with the duty of care: failure to follow up, failure to warn and failure to diagnose. The study found there is cause for concern about doctors' follow-up and documentation of patient care. Doctors may be unaware of the Australian courts' propensity to emphasise practitioner responsibility rather than patient autonomy. A further important result is the considerable disparity between the surgeons' views and the views of their practice managers about the duty of care. The article draws out implications for improved risk awareness and suggests further research. PMID- 14526727 TI - The law and practice associated with advance directives in Canada and Australia: similarities, differences and debates. AB - This article is a summary of research that investigated the Canadian and Australian legislative framework associated with advance directives in health care. The research focused on the context in which older people are encouraged to use advance directives. These are directions about refusal of medical treatment given in advance of incompetence. An advance directive may be given in a written document (living will) expressing one's wishes, by appointing another person (proxy) to make the decisions, or as a combination of the two. A lack of consistency and clarity about the terminology was found in both countries. This could be a barrier for older people to express their wishes in advance. Several confusing issues were also identified with the legislation related to advance directives. There appears to be a move towards appointing a substitute decision maker, but with significant differences across the Australian States and in Canadian Provinces. The "conversation" about future decisions emerged as an important theme, together with an emphasis on the process of "advance care planning" replacing the focus on advance directive forms. PMID- 14526728 TI - Private thoughts of public representatives: assisted death, voluntary euthanasia and politicians. AB - Assisted death and voluntary euthanasia have received significant and sustained media attention in recent years. High-profile cases of people seeking assistance to end their lives have raised, at least in the popular press, debate about whether individuals should be able to seek such assistance at a time when they consider their suffering to be unbearable or their quality of life unsatisfactory. Other recent developments include a number of attempts to legislate on the issue by the minor parties in Australia and the successful enactment of legislation in a few overseas jurisdictions. However, despite all of the recent attention that has focused on assisted death and voluntary euthanasia, a discussion of the adequacy of existing laws has not made it onto the political agenda of any of the Australian State or Territory governments. This is in spite of the fact that the private views of the majority of our elected Members of Parliament may be supportive of reform. This article explores the role of politicians' views and, as a case study, considers the opinions expressed by a number of Queensland Members of Parliament. In light of the views of these politicians and those of members of the public, as well as considerations arising from current medical practice, the article argues that there is a need for open political debate on assisted death and voluntary euthanasia. The article also suggests ways that such a debate may be achieved while minimising any political impact on governments that are prepared at least to consider this issue. PMID- 14526729 TI - The use of medical examinations for employment purposes. AB - This article examines a number of legal issues which arise where medical examinations or tests are used in the employment context, either to test the suitability of a prospective employee in pre-employment situations or to ascertain the fitness of existing employees. Employer justifications for seeking medical information usually relate to attempts to comply with health and safety legislation and to reduce workers compensation costs. This article discusses the legal obligations involved in pre-employment medical testing and, in particular, employee obligations to provide correct information in relation to their health to a prospective employer. It also notes the consequences for employees of providing false information in relation to workers compensation claims and dismissal cases. The article notes the growing trend of requiring drug and alcohol testing for existing employees, particularly in the mining industry. The article concludes by noting that employers need to take care in seeking medical information, and that it should not simply be done as a mechanical, matter-of course exercise. Failure to observe anti-discrimination laws may result in the employer facing allegations of misuse of medical information and claims for damages for discrimination. PMID- 14526732 TI - Exploiting genetic knowledge: the double helix 50 years on. Conference report from the United Kingdom. PMID- 14526730 TI - Diminished responsibility: the difference it makes. AB - Should there be a defence of diminished responsibility after the mandatory life sentence for murder has been abolished? This is the main question posed in this article, on the basis of a review of the central features of the defence and its surrounding forensic context. The views of the defence taken by the Law Reform Commissions of New South Wales and New Zealand are then compared. In conclusion, it is argued that opinions about the continuing validity of the defence of diminished responsibility are bound to be influenced by wider positions taken on the value of rules, as opposed to discretion, in the criminal law. PMID- 14526731 TI - Proof of cancer causation and expert evidence: bringing science to the law and the law to science. AB - This article discusses the medico-scientific and the legal views of cancer causation and how these two approaches impact on expert evidence. Cancer cause lends itself well to an exploration of the critical issues which surround its proof and the role of expert evidence in this proof. The article does not seek to identify or to resolve all the controversies or inconsistencies in the area. Rather, it sets up a basic framework for the general presentation and testing of expert medico-scientific evidence in litigation related to cancer causation. Specifically, it seeks to identify the technical questions of law and medical science regarding which medico-scientific disciplines can be relevant to proving cancer causation, and who the relevant expert witnesses to achieve this task would be. Emerging areas of cancer causation are then examined in light of this framework. PMID- 14526734 TI - Data pave the way to quality. PMID- 14526735 TI - Design solutions. Transforming Bethesda Lutheran Homes, Watertown, Wis. PMID- 14526733 TI - States tackle fragmented MR/DD systems. PMID- 14526736 TI - Achieving excellence in leadership. PMID- 14526738 TI - A challenge for you. PMID- 14526737 TI - When QA documents are privileged. PMID- 14526739 TI - Coding controversy. PMID- 14526740 TI - Change attitudes; save hospitals. PMID- 14526742 TI - Understanding when to use the new patient E/M codes. PMID- 14526741 TI - Spanning the global surgical package. PMID- 14526743 TI - Why did that idea flop? PMID- 14526744 TI - Creating a vital burn out-proof practice. PMID- 14526745 TI - Help with the HIPAA transactions and code sets standards. PMID- 14526748 TI - Prescription for progress. PMID- 14526747 TI - 8 ways to improve your relationships. PMID- 14526746 TI - Making decisions at the point of care: sore throat. PMID- 14526749 TI - The obesity pandemic. Is it beyond control? PMID- 14526751 TI - Polycystic ovaries: treatment strategies. PMID- 14526752 TI - Do we really know vitamin and mineral requirements for infants and children? AB - Infants and young children require vitamins and minerals not only to replace losses through metabolic turnover, but also to increase body reserves as they grow. For infants up to six months of age it is generally assumed that breast milk provides adequate intakes, and average breast milk composition provides the basis of reference intakes for this age group, and the basis for infant formula. It is questionable to what extent the micronutrient content of breast milk reflects the infant's requirements as opposed to the mother's nutritional status. For obvious ethical reasons there have been very few (if any) experiments to determine the micronutrient requirements of infants and children, although studies of intakes in areas where deficiency is common provide an estimate of minimum requirements in some cases. Different national and international authorities have approached the problem of estimating reference intakes for infants and young children in four main ways: linear interpolation between adequate intakes estimated from breast milk composition and the experimentally determined requirements of young adults; extrapolation forwards from breast milk composition and backwards from the requirements of young adults on the basis of metabolic body weight (body weight0.75), corrected by growth factors; extrapolation backwards from the requirements of young adults on the basis of the energy density of the diet; and by factorial calculation. Despite this, there is considerable concordance between the figures published by different authorities as a result of rounding off to avoid spurious precision. PMID- 14526750 TI - Highlights of the Health and Social Care (Community Health and Standards) Bill. PMID- 14526753 TI - Advances in the nutrition of preterm infants. AB - The greatly improved survival rate of infants born both preterm and low birth weight (LBW) has led to the subsequent growth and development of these infants becoming an important focus for research. Preterm infants begin life with, or acquire as a result of their prematurity, greater morbidity than term born babies, growth deficits, an increased risk of developmental delay and an increased risk of later adult diseases compared with appropriate for gestational age (AGA) term born babies. Research in recent decades has confirmed that there are marked differences in the nutritional requirements of preterm LBW infants compared with their AGA term born counterparts, both in the neonatal period and probably for all of infancy. In addition to the increased requirement for energy and protein, preterm LBW infants demonstrate a greatly increased requirement for some of the mineral elements, particularly iron, zinc and calcium, when compared with the needs of term AGA infants. In the UK, feeding practices for preterm infants in neonatal units and throughout infancy after hospital discharge are variable and many questions remain as to the optimal nutritional regimen for preterm LBW infants (and for subgroups of these infants) at different stages of infancy. There is some concern that the 2002 World Health Organization recommendations on infant feeding may be applied to all infants, including preterm infants, without consideration of their special nutritional needs, which may further compromise their growth and development. A brief resume of the work of prominent researchers in the field of preterm infant nutrition in the UK, notably Lucas, Cooke and Fewtrell, is included in the review, together with information from papers published by the authors of the review. The review concludes with a summary of the generally accepted recommendations on feeding preterm LBW infants after hospital discharge and information on some practical help available to the parents of these children and to health workers in the field. PMID- 14526754 TI - Some aspects of the public health importance of measurement of growth. AB - Growth is a fundamental process of life which in complex animals is influenced by genetic constitution, nurture and nutrition. Increase in size is accompanied by process of development and both growth and development are closely linked with time--the age of the individual. This paper considers ways in which simple measurements of different aspects of the growth of children have contributed, and are continuing to contribute, to our understanding of the origins of disease. The results of these measurements can help those involved in the care of children to identify individuals at risk of disturbances of growth and the possibility of the development of diseases associated with them. It can also help instigate preventative measures that may help children to develop more fruitful and longer disease-free lives. PMID- 14526755 TI - Is breast best? Is early solid feeding harmful? AB - The health benefits of breastfeeding for babies and mothers have long been recognised and it is now globally recommended that it be continued exclusively for six months. Although there are few controlled trials to support this recommendation, the most important advantage is less morbidity from gastrointestinal infection in developing countries. There is also evidence that respiratory tract infections and atopic dermatitis is reduced, and the maternal risk of breast cancer decreases, particularly with a longer duration of breastfeeding and a high parity. There is little to suggest that exclusive breastfeeding for six months adversely affects infant growth, nutritional status or infant feeding skills, but more studies are needed. Equally, there is no evidence that introduction of solids from 17 weeks is harmful in developed countries. However, in the UK breastfeeding prevalence is low and solids are introduced early for the majority of infants and much can be done to positively encourage and support all mothers to continue breastfeeding for a longer period. PMID- 14526756 TI - Estimation of the impact on children's health of environmental tobacco smoke in England and Wales. AB - In this paper, the population attributable risk (PAR), a measure of the excess risk of disease associated with a risk factor, is calculated for some of the common adverse health effects that have been associated with exposure of children to environmental tobacco smoke (ETS): childhood lower respiratory illness, chronic middle ear disease, asthma and sudden infant death syndrome (SIDS). Published data on both risk estimates and the percentage of children exposed to ETS in the home (prevalence of ETS) have been utilised. The percentage of childhood lower respiratory illness and middle ear disease typically attributable to ETS from either parent smoking ranged from 9% for asthma prevalence and for referral for glue ear, to 25% for hospital admission for lower respiratory illness. Where data were available to calculate PARs separately for mother only smoking and father only smoking, the PARs were generally larger for mothers only smoking, due mainly to higher odds ratios for mothers only smoking. The PAR for SIDS attributable to ETS from mother only smoking was 11%. Although based on a small number of studies, the PAR for SIDS attributable to smoking of fathers only was similar to that attributable to the smoking of mothers only, largely due to the higher prevalence of households where only the father smokes. This study has shown that the impact of ETS on childhood illness can be considerable, emphasising the importance of the need to develop effective strategies for reducing the risk of ETS exposure in the home and elsewhere. PMID- 14526757 TI - Lessons to be learned: a case study approach. Chronic intussusception in childhood. AB - Chronic intussusception as a cause of persistent abdominal pain in children is often an overlooked diagnosis. Here we present an eight-year-old boy, who at the age of three years had an acute intussusception reduced hydrostatically with barium and who subsequently had been extensively investigated both in Wales and in Switzerland, for persistent colicky abdominal pain. He was found to have chronic intussusception, with a Meckel's diverticulum being the cause of his symptoms. PMID- 14526759 TI - [German version of the Strength and Difficulties Questionnaire (SDQ-German)- overview and evaluation of initial validation and normative results]. AB - The Strengths and Difficulties Questionnaire (SDQ) is a short behavioural screening questionnaire which can be completed in about five minutes by parents and teachers of 4- to 16-year-olds or as self-report by 11- to 16-year-olds. The English original has already been fully evaluated and is widely used in research and clinical practice. The instrument was translated into German in 1997, and several evaluative studies have since been completed. The present paper gives an overview of this novel instrument and summarizes the normative and validation studies to date. A normative study on a field sample of 930 children demonstrated that the distributions of raw scores in the German parent SDQ closely resemble those found in the English version, while a factor analysis of the German data yielded a pattern of loadings which convincingly replicated the original scale structure. Initial validation studies showed that the parent-, teacher-, and self completed SDQ-Deu correlates well with the considerably longer German versions of the Child Behavior Checklist (CBCL) and respective teacher and self-report derivatives (TRF, YSR). Both parent-rated instruments are equally able to distinguish between a community and a clinic sample, and between subgroups with and without specified categories of disorders within a clinic sample. After discussing possible uses of the SDQ-Deu as well as similarities and differences to other scales, we conclude that the German SDQ is just as useful and valid an instrument for many clinical and research purposes as the English original. PMID- 14526758 TI - [Development and evaluation of a psychoeducational parent group training program for families with autistic children]. AB - In the introduction of this study, the concept of "psychoeducational parent training" is analyzed, and an outline of the research literature on "parent training in families with autistic children" is presented. Based on these findings a psychoeducational group parent training, which focused on (1) addressing issues of nature, etiology, treatment and family-related consequences of autism (2) teaching child management and education skills was developed, and the training outcomes were evaluated within a 3-months-follow-up design. A total of 24 parents of 23 autistic children (mean age: 9 yrs.) participated in the center-based training program which was performed in three small groups in different areas of Germany in 3 one-day sessions succeeding in 1-month-intervals. The outcome variables included: (a) group training assessments by parents; they evaluated quality of trainer variables, curriculum, and group atmosphere using a questionnaire format of bipolar rating scale-items (b) parental 3-months-follow up assessments of the effects of group training on parent-child interactions and family adaptation, using a questionnaire based on rating scales, and a semi structured questionnaire on training-related child and parent behaviors in the family. The parent training resulted in (a) a high degree of parent satisfaction with the training format and (b) positive effects on daily parent-child interactions from the perspective of parents. These findings provide some evidence for both clinical and social validity of the parent training procedure examined in this study. PMID- 14526761 TI - [Planning and implementation of evaluation studies in child and adolescent psychiatry]. AB - An introduction into planning and implementing evaluation studies in child and adolescent psychiatry is presented. After the presentation of theoretical aspects of evaluation studies, the necessity of these studies is elucidated. Further, a conceptualisation of the dimensions of a treatment programme and some criteria for the selection of outcome criteria are offered. In addition to symptomatology, other criteria such as family functioning and quality of life are proposed in this context. Furthermore, it is recommended to ascertain data already at the time of the patient's registration. The influence of drop-out rate on the study result makes it necessary to closely consider any missing data. These and other problems lead to the conclusion that a sophisticated and complex evaluation methodology is required to assess the performance of a child and adolescent treatment programme validly. Finally, the design of the Hamburg evaluation study is presented, which is suitable for the implementation of some of the mentioned aspects. PMID- 14526762 TI - Johnson & Johnson campaign helping to reduce nursing shortage. PMID- 14526760 TI - [Factors influencing the course and duration of inpatient child and adolescent psychiatric treatment: between empiricism and clinical reality]. AB - In the last two decades considerable changes influenced the scope of inpatient treatment in child and adolescent psychiatry. Proceeding from a literature review dilemmas between available research data and clinical practice will be pointed out. Proposals will be made to take into account the complex developmental processes, the individuality and the social context by psychic impaired children and adolescents requiring hospitalisation. This could improve the transfer of research findings into the clinical practice. It will be argued against a confusion of economical interests with research findings. PMID- 14526763 TI - Impact of random and fixed (optimal) sampling approach on the Bayesian estimation of clearance. AB - BACKGROUND AND OBJECTIVE: The population pharmacokinetic approach is based on sparse sampling. In sparse sampling approaches, the selection of the time point(s) is very critical for the prediction of pharmacokinetic parameters. Several investigators have shown that the predictive performance of the Bayesian approach is influenced by the initial estimates of pharmacokinetic parameters as well as the time of blood sampling. The objective of this study is to evaluate the impact of random and fixed sparse sampling approach on the Bayesian estimation of clearance. METHODS: Three drugs were selected for this study. Two sparse sampling methods (random or fixed) using Bayesian approach were used to assess clearance in healthy subjects following a single oral dose. The initial estimates of the model parameters and inter- and intra-subject variabilities were obtained from the previous pharmacokinetic studies conducted in healthy volunteers. The predicted clearance values using sparse sampling (1, 2 or 3 blood samples per subject) were compared with the clearance values obtained by extensive sampling. RESULTS AND CONCLUSION: The results indicated that both random and fixed sampling approaches, irrespective of number of blood samples, can be used to estimate mean population as well as post hoc predicted individual clearance (Bayesian) with accuracy. However, the precision of the prediction of clearance was found to be better with fixed rather random blood sampling approach. PMID- 14526765 TI - The roles of fatalism, self-confidence, and intellectual resources in the disablement process in older adults. AB - In this study, the authors examined the relations between 3 psychological variables-fatalism, self-confidence, and intellectual resources-and the subsequent development of illness and disability 20 years later in an adult sample. Results indicated that greater fatalism, assessed in 1974, predicted greater difficulty in everyday cognitive tasks as well as illness in 1994. Higher self-confidence in 1974 was associated with lesser degrees of cognitive and fine motor difficulty in 1994. Greater intellectual resources in 1974 (a combination of intellectual flexibility and education) predicted less cognitive and gross motor difficulty as well as lesser degrees of illness in 1994. Some of these relations were stronger for older than for middle-aged individuals. Results are discussed in the context of models of the disablement process. PMID- 14526766 TI - Comment on: Tani M, Sakai P, Kondo H. Endoscopic mucosal resection of superficial cancer in the stomach using the cap technique. Endoscopy 3003;35:348 355. PMID- 14526764 TI - Mitochondrial DNA depletion and respiratory chain enzyme deficiencies are present in peripheral blood mononuclear cells of HIV-infected patients with HAART-related lipodystrophy. AB - The main objective of the present study was to ascertain if mitochondrial DNA (mtDNA) depletion as reported in HIV-infected patients with highly active antiretroviral therapy (HAART)-related lipodystrophy (LD) implies any degree of mitochondrial respiratory chain (MRC) dysfunction. For this purpose, we evaluated HIV patients on different HAART schedules with LD (group A; n=12) and on HAART but without LD (group B; n=12), and untreated HIV-infected patients as controls (group C; n=24). mtDNA content was determined on peripheral blood mononuclear cells (PBMCs) with a real-time PCR method. Complex II, III and IV activities of the MRC were simultaneously measured spectrophotometrically, as were spontaneous and stimulated oxygen consumption by PBMCs. Compared to controls (group C, 100%), patients with LD (group A) showed a decreased mtDNA content (54%, P<0.001), which was associated with a decline in complex III (62%, P<0.05) and IV activity (69%, P<0.05) (both complexes partially encoded by mtDNA), but not in complex II activity (exclusively encoded by nuclear DNA). Patients in group B showed a similar pattern of mitochondrial dysfunction but to a lesser extent and without statistical significance. Respiratory activities in both treated groups (A and B) did not differ in comparison with controls. We conclude that mtDNA depletion occurring during HAART is associated with deficiencies in MRC complexes partially encoded by mtDNA, which are detectable by PBMCs. Presented in 'Late Breakers and Hot Topics' session at 6th International Congress on Drug Therapy in HIV Infection, Glasgow, UK, 17-21 November 2002. PMID- 14526767 TI - Dalteparin compared with an oral anticoagulant for thromboprophylaxis in patients with cancer. PMID- 14526768 TI - Dalteparin compared with an oral anticoagulant for thromboprophylaxis in patients with cancer. PMID- 14526769 TI - Dalteparin compared with an oral anticoagulant for thromboprophylaxis in patients with cancer. PMID- 14526770 TI - Cardiovascular genomics. PMID- 14526771 TI - Cardiac transplantation in an HIV-1-infected patient. PMID- 14526772 TI - Master in medical science with major in neuroradiology. PMID- 14526773 TI - Response to article by Zetterstrom et al. PMID- 14526774 TI - Omission of interferon alpha information in Nelson at al.'s article. PMID- 14526775 TI - United Nations panel warns of slow progress against HIV/AIDS. Current levels of funding and programme implementation insufficient to meet "greatest challenge of our time". PMID- 14526776 TI - Can cities be designed to fight obesity? Urban planners and health experts work to get people up and about. PMID- 14526777 TI - WHO targets measles and hepatitis B in Western Pacific. PMID- 14526778 TI - Dutch government announces large cuts in health spending. PMID- 14526779 TI - Concern over mercury pollution in India. PMID- 14526780 TI - More cash for health care in China. PMID- 14526781 TI - Drug-resistant strains of HIV increase sharply in Japan. Health officials step up efforts to improve diagnosis and ensure adherence to treatment. PMID- 14526782 TI - Bleeding disorders. Von Willebrand factor. PMID- 14526783 TI - [The Vilnius Gynecological Society in years 1925-1939]. AB - In this paper, the history of the Vilnius Gynecological Society is presented. The Society was founded in autumn 1925 and was active until September 1939. The origin of the organisation, its beginnings, development and rules of functioning are presented here. The complete list of members of the Society (1925 - 1939) is enclosed. PMID- 14526784 TI - [The life of Ludwik Rydygier in a village of Grabowo]. AB - There have been a few biography notes concerning Prof. Ludwik Rydygier's personal life, as well as professional record. Most of them has concentrated on study period and following professional activity. On the other hand, there is very little information concerning his childhood and first years of his elementary education. The name of Grabowo has been quite recently found as location connected with the great surgeon. Rydygier's family had been living there about 22 years. Grabowo was a place of birth and baptism of the six of the surgeon's younger brothers and sister. From Grabowo Ludwik Rydygier was leaving to schools and for the university. PMID- 14526785 TI - ["Decemberal" crisis in Polish Society of History of Medicine]. AB - In December 1970 in Warsaw the meeting of the Polish Society of History of Medicine delegates was performed. The Author who was the Lower Silesian section delegate described the course of this meeting with his personal point of view. The programme's main point was the report of former Board and the election of new one. The discussion during the meeting concentrated on the polemic between former chairman Prof. Stanislaw Konopka and his successor Doc. Stanislaw Szpilczynski from Wroclaw. The very fierce discussion was moderated by the meeting's president prof. Kazimierz Lejman from Krakow. The Author calls the attention to the accidental coincidence of the date of the meeting and the political crisis in Poland, caused by the strikes in the Balticseaside. PMID- 14526786 TI - Meningogoccal meningitis. PMID- 14526787 TI - A simple but ambitious plan: 20 million dollars to jump start the Iraqi health system. PMID- 14526788 TI - World Health Assembly adopts historic tobacco control pact. PMID- 14526789 TI - Has the history of Canadian medicine come of age? A personal view. AB - This short "think piece" interrogates the present state of medical history in Canada. What, I ask, are the past accomplishments and future prospects of this new genre of historical writing in Canada? Has Canadian medical history come of age? And what do we mean by asking this question? My answer is that while there is much to celebrate in the sterling individual works in the field in the past 20 years, the field more generally has yet to confront more difficult questions of identity, theory and methodology that, in my view, any young subfield needs to tackle to survive in the presently overcrowded academic marketplace of approaches and ideas. PMID- 14526790 TI - Tribute to Charles Roland. PMID- 14526791 TI - [Ahern, historian of medicine]. AB - Michel Joseph Ahern began the historical study of the medical profession in Quebec in 1910, with the publication of a series of biographies entitled Notes pour servir a l'histoire de la medecine dans le Bas-Canada depuis la fondation de Quebec jusqu'au commencement du XIXe siecle in the Bulletin medical de Quebec. After his death, his son George used his extant notes to continue the publication of these texts and a subsequent one-volume compilation with the same title. Despite several weaknesses, this work remains an indispensable source for researchers interested in the medical history of Quebec. Prepared in the context of several commemorations at the beginning of the 20th Century such as the 300th anniversary of the founding of Quebec City, Ahern's texts were tributes to the founders of medicine in Quebec, irrespective of their status; consequently, this work is not a pantheon of great names in medicine in the period. Doctor Ahern strove for historical integrity, and consulted archives, historians, and the learned men o f his time. PMID- 14526793 TI - [The history of health in Quebec: relationships and specificities]. AB - The remarkable development of the history of health in Quebec during the last two decades occurred at the junction of two main historiographical movements: the international resurgence of the history of health, and the metamorphosis of the historian's gaze toward Quebec. The author examines specific features issued from this junction. He shows that many historians challenged the "retard du Quebec" thesis. Studying, as they did, the relations between health service providers, they unveiled their respective interests and conflicts, which led to the invalidation of the idea of a monolithic backwards society. PMID- 14526792 TI - How much gin in the tonic? The problems of writing a provincial medical history,. AB - This paper describes the problems and resources involved in writing a Canadian provincial medical history, (Manitoba Medicine: A Brief History). The first decision was whether it should be a scholarly or a popular history; The authors' background, and the realities of publishing dictated the latter. Resources available were local and easily accessible: archives and records, the Manitoba medical journals, a series of local medical journals (almost continuous for a century), and the Manitoba medical biographies, books variable in length, and content, but relating to a wide variety of physicians. Such a paper leads to a question- "Is local history merely trivial?" The answer to such a question is "no." PMID- 14526795 TI - Harvard symposium on the clinical efficacy and hemostatic mechanism of action of poly-N-acetyl glucosamine. PMID- 14526796 TI - WHO steps up fight against poliomyelitis. PMID- 14526797 TI - AstraZeneca postpones discontinuation of Mysoline. PMID- 14526798 TI - Detection of cognitive impairment: the final frontier. PMID- 14526799 TI - Mapping "region" in Canadian medical history: the case of British Columbia. AB - The notion of "region" can be a valuable analytical tool in the writing of Canadian medical history. This article explores themes in the history of British Columbia that link medicine and regional development. Employing a historiographical sweep from the colonial period to the 1970s, the author considers doctors and imperialism, medical practice and the economy, and the relationship between metropolis and periphery in shaping medical institutions and medical culture in the western province. The intent of the piece is to stimulate thought about the potential of introducing the sense of place into regional medical history in Canada. PMID- 14526800 TI - Portraits of people with mental disorders in English Canadian history. AB - This paper will discuss the ways in which academic and non-academic historians in English Canada have portrayed people with mental disorders. The phrase "people with mental disorders" refers to individuals who were considered mentally disturbed by their contemporaries and/or by historians. This includes discussion of people who were in mental institutions as well as people who were never confined. Citations are from both detailed studies and fleeting references on this topic. Areas considered include portraits of people with mental disorders as being anonymous, passive, active, named, dangerous, untrustworthy; they also include the problem of stereotypes in the writing of this history. PMID- 14526801 TI - Early settlements in Newfoundland and the scourge of scurvy. AB - Two important archeological investigations currently underway in the Avalon Peninsula of Newfoundland, at Cupids (established 1610) and Ferryland (1621), help to focus questions on health and health care in early seventeenth-century English settlements. This review looks at one aspect only, the problem of scurvy and what treatments were tried. In adding to our understanding of early colonial settlements and on how new medicinal plants were "discovered," the Newfoundland story reveals a considerable interest in local plants as substitutes for scurvy grass. Suggestions are offered on how they may have been chosen and the rationales behind their use. PMID- 14526802 TI - "Take up thy bed and walk": Aimee Semple McPherson and faith-healing. AB - Aimee Semple McPherson was an important factor in the spread of Pentecostalism in the first half of the 20th century. Although she always identified herself as Canadian, she built her temple in Los Angeles, from which she hoped to evangelize the world. Before settling down in 1923, however, she experienced unnerving success as a faith-healer. Never comfortable with the role, not least of all because she was committed to saving souls, not bodies, Sister McPherson attempted during her secluded final decade to devise a theory of faith-healing. Although still celebrated by her followers as a prophet and a healer, she had expressed discomfort in her lifetime with both these roles. Physically, she ran herself into the ground and died at the age of 54 from a drug overdose. PMID- 14526803 TI - Poisoning the spindle: serendipity and discovery of the anti-tumor properties of the Vinca alkaloids. AB - In 1995, Canadian scientists Robert Noble and Charles Beer were inducted into the Canadian Medical Hall of Fame for their 1950s "discovery" of Vinblastine. Their "chance" finding of an anticancer drug in the leaves of the periwinkle plant (Vinca rosea, Linn.), is used to explore the historical issue of discovery, accidental discovery, and priority. The elements of the discovery are reconstructed through the oral testimony of key players and their published and unpublished records. Several "unsung heroes" played key roles in this project and reasons for their relative invisibility will be presented. Special attention is paid to the relationship between the small Canadian academic group working at UWO and the large pharmaceutical company (Eli Lilly) engaged in similar research at the same time. PMID- 14526805 TI - "Lively combat": Kathleen Ellis and the Canadian Nurses Association's lobby during the Second World War. AB - Although it has become unfashionable in recent years to extol the achievements of those who have become known as the "elite" in Canadian nursing, it is nonetheless true that a number of early Canadian nurse leaders contributed significantly to the advancement of their profession. One such individual was Kathleen Wilhelmina Ellis. This article analyzes her impact as Emergency Nursing Advisor for the Canadian Courses Association (CNA) during World War II. The previously unreported success of the CNA in capitalizing on the "crisis" of world War II to counter plans of the Canadian Hospital Council to increase pupil nurse enrollments beyond hospitals' clinical teaching and supervision capabilities provides the context for analysis. From 1942 to 1946, the CNA administered a $774,000 grant from the Federal Government to improve teaching in hospital and university schools of nursing, provide bursaries for graduate nurses, and recruit increased numbers of qualified applicants to hospital nurse training programs. The success of the CNA in securing and administering these funds signalled its political acumen as national representative of Canadian nurses and nursing. Kathleen Ellis' contribution as Emergency Nursing Advisor for the CNA was important to the success of the CNA's lobby during World War II. PMID- 14526806 TI - The other branch of medicine: an historiography of veterinary medicine from a Canadian perspective. AB - Animal (veterinary) medicine, that other branch of medical healing, forms a part of medical history and there are themes in common between human and veterinary medical historiography. Some of these themes include "great docs, great diseases," struggles with professionalization, institutional stories, general chronological overviews, development of medical disciplines, and the role of women. The "patient's view" in veterinary historiography is somewhat different. Veterinary history includes the theme of "great animals" but this is not, in the medical historical sense, a "patient's" view. Much work in this area of Canadian history remains to be accomplished. PMID- 14526804 TI - The sometimes uncertain world of Canadian obstetrics, 1900-1950. AB - All specialties in medicine in the first half of the twentieth century partook of the prestige of medical science and their practitioners looked to the past as a gauge to how far their specialty had progressed. Obstetrics was no different. The introductions to most obstetric textbooks and articles contained almost mandatory praise for the advances made. The writers acknowledged the art of obstetrics and, at times, gave it equal rhetorical standing, but in emphasis, the "science" of obstetrics dominated. Yet underlying the bravado was an unease that not all was well with the field. Several issues contributed to the sense of anxiety-the deficiencies in obstetrical education, its status vis-a-vis other medical specialties, the reality of practice, including the remuneration received, and the perception of competition from "unqualified" individuals. Such concerns are important for historians of childbirth to recognize. PMID- 14526808 TI - The archives of the Canadian Dermatology Association. AB - This article describes some details of the archival material of the Canadian Dermatology Association (CDA). Covering its 75 years of existence, these archives consist of seven items: (1) 38 volumes of bound archives; (2) tri-annual issues since 1975 of the CDA Journal/Bulletin (a socio-political historical newsletter); (3) members' files containing extensive biographical material; (4) a collection of 150 books and other pamphlets; (5) since 1995, the scientific peer-reviewed official journal of the CDA (Journal of Cutaneous Medicine and Surgery); (6) a sporadic collection of photographs, meeting tape recordings, and protocols of patient presentations at the annual meetings; and (7) a miscellany of historical articles on Canadian dermatologists, dermatology published in older medical Journals, and two books published separately. PMID- 14526807 TI - The casebook, the daybook, and the diary as sources in medical historiography. AB - Casebooks, daybooks and diaries were among a number of literary forms commonly used by physicians in the nineteenth century to record the details of their medical cases. Because they constitute primary sources on medical practice in that era they have value to the medical historian. To illustrate this, the writings of three Canadian physicians' -John Mackieson, Jonathan Woolverton, and James Langstaff- are discussed, together with a consideration of some of the principles relating to the study of their manuscripts, particularly in relation to the study of the history of practice in rural areas. PMID- 14526809 TI - History of the Canadian Society for the History of Medicine. PMID- 14526810 TI - James Rachels, ethicist, 62; ignited euthanasia debate. PMID- 14526812 TI - When a doctor stumbles on a family secret. PMID- 14526811 TI - Emergency rooms get eased rules on patient care. PMID- 14526814 TI - Future perfect? Britain is in the vanguard of genetics research, and the government wants to put this to practical use. PMID- 14526813 TI - Senate makes a curb on abortion likely. PMID- 14526815 TI - To sell pricey drug, Lilly fuels a debate over rationing. PMID- 14526816 TI - State to give liver donors safeguards in transplants. PMID- 14526817 TI - Science journals tighten rules for disclosure of financial ties. PMID- 14526818 TI - Son's wish to die, and mother's help, stir French debate. PMID- 14526819 TI - Naval center halts research during inquiry about ethics. PMID- 14526822 TI - Do you have misteaks [sic] in your resume? PMID- 14526820 TI - British women lose court fight over possessing frozen embryos. PMID- 14526821 TI - The Resuscitation greats. J D Herholdt and C G Rafn: two unsung heroes from Denmark. PMID- 14526823 TI - Measured and modeled radiometric quantities in coastal waters: toward a closure. AB - Accurate radiative transfer modeling in the coupled atmosphere-sea system is increasing in importance for the development of advanced remote-sensing applications. Aiming to quantify the uncertainties in the modeling of coastal water radiometric quantities, we performed a closure experiment to intercompare theoretical and experimental data as a function of wavelength lambda and water depth z. Specifically, the study focused on above-water downward irradiance E(d)(lambda, 0+) and in-water spectral profiles of upward nadir radiance L(u)(lambda, z), upward irradiance E(u)(lambda, z), downward irradiance E(d)(lambda, z), the E(u)(lambda, z)/L(u)(lambda, z) ratio (the nadir Q factor), and the E(u)(lambda, z)/E(d)(lambda, z) ratio (the irradiance reflectance). The theoretical data were produced with the finite-element method radiative transfer code ingesting in situ atmospheric and marine inherent optical properties. The experimental data were taken from a comprehensive coastal shallow-water data set collected in the northern Adriatic Sea. Under various measurement conditions, differences between theoretical and experimental data for the above-water E(d)(lambda, 0+) and subsurface E(d)(lambda, 0-) as well as for the in-water profiles of the nadir Q factor were generally less than 15%. In contrast, the in water profiles of L(u)(lambda, z), E(d)(lambda, z), E(u)(lambda, z) and of the irradiance reflectance exhibited larger differences [to approximately 60% for L(u)(lambda, z) and E(u)(lambda, z), 30% for E(d)(lambda, z), and 50% for the irradiance reflectance]. These differences showed a high sensitivity to experimental uncertainties in a few input quantities used for the simulations: the seawater absorption coefficient; the hydrosol phase function backscattering probability; and, mainly for clear water, the bottom reflectance. PMID- 14526825 TI - Detection and false-alarm probabilities for laser radars that use Geiger-mode detectors. AB - For a direct-detection laser radar that uses a Geiger-mode detector, theory shows that the single-pulse detection probability is reduced by a factor exp(-K), where K is the mean number of primary electrons created by noise in the interval t between detector turn-on and arrival of laser photons reflected from the target. The corresponding false-alarm probability is at least 1 - exp(-K). For fixed-rate noise, one can improve the detection and false-alarm probabilities by reducing t. Moreover, when background-light noise is significant and dominates dark-current noise and when the laser signal is of the order of ten photoelectrons or more, the probabilities can be improved by reducing the amount of light falling on the detector, even if the laser signal is reduced by the same factor as the background light is. Additional analytical calculations show that identifying coincidences in data from as few as three pulses canreduce the false-alarm probability by orders of magnitude and, for some conditions, can also improve the detection probability. PMID- 14526824 TI - Asymptotic optical depths in source-free ocean waters. AB - The depth dependence for which the downward diffuse attenuation coefficient, the upward-to-downward plane irradiance ratio, the vertically upward radiance-to downward plane irradiance ratio, and the mean cosine of the radiance depend negligibly on the surface incident illumination have been examined. The depths at which these coefficients approach to within a specified percent of their asymptotic values depends significantly on the characteristics of the incident illumination and on the inherent optical properties of the water. This information is useful when solving inverse ocean optics problems with a method for which the radiance is assumed to be approximately in the asymptotic regime. PMID- 14526826 TI - Design, implementation, and characterization of a folded spot-array generator for a modulator-based free-space optical interconnect. AB - A folded structured light generator is presented. This spot generator is to be used in a modulator-based free-space optical interconnect. Three cascaded diffractive optical elements produce 4 x 8 clusters on an 800 microm x 1600 microm pitch, in which each cluster is a 4 x 4 array of 13.1-microm-radius spots on a 90-microm pitch. The folded configuration is more compact than an existing linear spot-array generator and replaces 14 optical surfaces with eight surfaces. Details of the optical design, sensitivity analysis, alignment techniques, assembly, and test results are presented. PMID- 14526827 TI - In-line liquid-crystal microcell wave plates and their application for high speed, reset-free polarization mode dispersion compensation in 40-Gbit/s systems. AB - We describe the design, fabrication, and performance of a high-speed, continuously tunable, and reset-free polarization controller based on nematic liquid-crystal (NLC) microcell wave plates fabricated directly between the tips of optical fibers. This controller utilizes a pulsed driving scheme and optimized NLC materials to achieve a stepwise switching speed of 1 deg/micros, for arbitrary rotation angles with moderately low voltages. This compact microcell design requires no bulk optical components and has the potential to have low insertion loss. We describe the performance of these devices when implemented in polarization mode dispersion compensators for 40 Gbit/s systems. The good optical properties and the nonmechanical, high-speed, and low-power operation suggest that this type of device might be considered for some applications in dynamic compensation of polarization mode dispersion, polarization analysis, polarization division demultiplexing, and polarization scrambling in high-speed optical communication systems. PMID- 14526828 TI - Fiber-optic cavity sensing of hydrogen diffusion. AB - A novel type of fiber-optic cavity sensor for hydrogen diffusion into and out of fibers is presented. The sensor is an implementation of a cavity ringdown scheme in a silica-based single-mode fiber that has been exposed to gaseous hydrogen at normal pressure. The measured ringdown times during the H2 diffusion show good agreement with a theoretical diffusion model. This model allows the determination of the diffusion coefficient of hydrogen in silica, resulting in D = (3.02 +/- 0.07) x 10(-15) m2/s at 30 degrees C. PMID- 14526830 TI - Assessment of the accuracy of snow surface direct beam spectral albedo under a variety of overcast skies derived by a reciprocal approach through radiative transfer simulation. AB - With radiative transfer simulations it is suggested that stable estimates of the highly anisotropic direct beam spectral albedo of snow surface can be derived reciprocally under a variety of overcast skies. An accuracy of +/- 0.008 is achieved over a solar zenith angle range of theta0 < or = 74 degrees for visible wavelengths and up to theta0 < or = 63 degrees at the near-infrared wavelength lambda = 862 nm. This new method helps expand the database of snow surface albedo for the polar regions where direct measurement of clear-sky surface albedo is limited to large theta0's only. The enhancement will assist in the validation of snow surface albedo models and improve the representation of polar surface albedo in global circulation models. PMID- 14526829 TI - Information theoretical optimization for optical range sensors. AB - Most of the known optical range sensors require a large amount of two-dimensional raw data from which the three-dimensional (3D) data are decoded and so are associated with considerable cost. The cost arises from expensive hardware as well as from the time necessary to acquire the images. We will address the question of how one can acquire maximum shape information with a minimum amount of image raw data, in terms of information theory. It is shown that one can greatly reduce the amount of raw data needed by proper optical redundancy reduction. Through these considerations, a 3D sensor is introduced, which needs only a single color (red-green-blue) raw image and still delivers data with only approximately 2-microm longitudinal measurement uncertainty. PMID- 14526831 TI - Theory of quarter-wave-stack dielectric mirrors used in a thin fabry-perot filter. AB - I present a new derivation of the analytic form for the phase shift near resonance and the optical penetration length upon reflection from a distributed dielectric mirror consisting of a quarter-wave stack. The requirement of proper termination to achieve high reflectivity is suspended to investigate large optical penetration depths. Separate equations, derived for N and N + 1/2 layer pairs, are convenient for the design of tunable Fabry-Perot filters with a specified tuning range. The analysis is also applicable to distributed Bragg reflectors, vertical-cavity surface-emitting lasers, and resonant photodiodes. I show that the penetration length can sharply reduce the overly broad free spectral range of an ultrathin Fabry-Perot filter that might be useful in applications such as tunable wavelength filters for wavelength division multiplexing applications. The results also demonstrate regimes of zero dispersion and of superluminal reflection in the dielectric mirrors, which are of particular interest in photonic bandgap structures. PMID- 14526832 TI - Fabrication of two-dimensional photonic crystals with controlled defects by use of multiple exposures and direct write. AB - We have developed an approach for relatively rapid and easy fabrication of large area two-dimensional (2-D) photonic crystal structures with controlled defects in the lattice. The technique is based on the combination of two lithographic steps in UV-sensitive SU-8 photoresist. First, multiple exposures of interference fringes are used in combination with precise rotation of the sample to define a 2 D lattice of holes. Second, a strongly focused UV laser beam is used to define line-defect waveguides by localized exposure in the recorded but not yet developed lattice from the first step. After development, the mask is transferred into a GaAs substrate with dry etching in chemically assisted ion-beam etching. PMID- 14526833 TI - Two-wavelength, passive self-injection-controlled operation of diode-pumped cw Yb doped crystal lasers. AB - We demonstrate and investigate a peculiar mode of cw Yb3+-doped crystal laser operation when two emissions, at two independently tunable wavelengths, are simultaneously produced. Both emissions are generated from a single pumped volume and take place in either a single beam or spatially separated beams. The laser employs original two-channel cavities that use a passive self-injection-locking (PSIL) control to reduce intracavity loss. The advantages of the application of the PSIL technique and some limitations are shown. The conditions for two wavelength multimode operation of the cw quasi-three-level diode-pumped Yb3+ lasers and the peculiarity of such an operation are carried out both theoretically and experimentally. The results reported are based on the example of a Yb3+:GGG laser but similar results are also obtained with a Yb3+:YAG laser. The laser operates in the 1023-1033-nm (1030-1040-nm) range with a total output power of 0.4 W. A two-wavelength, single longitudinal mode generation is also obtained. PMID- 14526834 TI - Linewidth reduction and wavelength tuning of an erbium-doped fiber laser by use of a single-mode-selected and side-mode-suppressed Fabry-Perot laser diode. AB - A single-mode and highly side-mode-suppressed 1.55-microm Fabry-Perot laser diode (FPLD) is achieved by feedback injection with an erbium-doped fiber laser (EDFL). For selection of the strongest longitudinal mode from the gain spectrum of the FPLD for lasing in the EDFL, the FPLD is operated at just below the threshold condition and is feedback injected by 0.02% of the EDFL output power. The lasing mode and center wavelength of the proposed single-mode FPLD source are decided by cross-correlated gain profiles of the EDFL and the FPLD; however, the effect of FPLD injection modes is found to be more pronounced. The optimized lasing linewidth (system limitation) and side-mode suppression ratio of 0.01 nm and > 49 dB are obtained, which are far better than those of a FPLD at free-running condition. The worst linewidths at 3- and 10-dB decay are observed to be at approximately 0.016 and 0.05 nm, respectively. Linear wavelength tuning of as much as 4.5 nm (from 1558.7 to 1563.2 nm) by adjustment of the temperature of the FPLD from 10 degrees C to 40 degrees C at just below threshold is reported. The wavelength-tuning slope is approximately 0.14 nm/degrees C under temperature accuracy of 0.1 degrees C. PMID- 14526835 TI - Laser-induced damage in deuterated potassium dihydrogen phosphate. AB - Laser-induced pinpoint bulk damage of deuterated potassium dihydrogen phosphate at 351 nm is shown to depend on the propagation direction relative to the crystallographic axes and on growth temperature in addition to the previously reported dependence on continuous filtration. Pulse-length scaling is also consistent with earlier reports. The leading hypothesis for the cause of pinpoint damage is absorbing nanoparticle impurities, and our results are consistent with but not conclusive for that model. Advances in technology have led to greatly improved damage resistance. PMID- 14526836 TI - Diode-pumped passively mode-locked multiwatt Nd:GdVO4 laser with a saturable Bragg reflector. AB - We report a first demonstration, to our knowledge, of a cw passively mode-locked Nd:GdVO4 laser (k = 1063 nm). A relaxed saturable Bragg reflector was used. The laser generates pulses of 9.2 ps at a repetition rate of 119 MHz. As much as 5.4 W of average power was realized with a slope efficiency of 25.7%. PMID- 14526837 TI - High-power 457-nm light source by frequency doubling of an amplified diode laser. AB - We demonstrate the generation of 150-mW blue coherent single-mode radiation at 457 nm in a compact and inexpensive setup. The light is generated by frequency doubling the radiation of a master oscillator power amplifier (MOPA) system in an enhancement cavity with a potassium niobate (KNbO3) crystal. The MOPA consists of a 914-nm single-mode diode laser and a broad-area diode laser (BAL) as the amplifier. The BAL is a multimode laser with a specified wavelength of 938 nm. Sufficient gain at 914 nm is obtained by antireflection coating the BAL front surface and by cooling it to -10 degrees C. PMID- 14526838 TI - Extremely uniform angular distributions of the three-dimensional emission spectra of photonic quantum ring lasers. AB - We have observed the emission spectra of three-dimensional Rayleigh-Fabry-Perot cavity whispering-gallery modes from photonic quantum ring lasers that give rise to uniform angular mode spacing in space. The observed angular distribution (0 degrees approximately 30 degrees) of the emission modes is linear with respect to the mode number. For example, the average angular mode spacing is 0.95 degrees with a standard deviation of 0.098 degrees for a 20-microm-diameter laser, whose potential angle-resolving power is promising for angle-sensing applications in the future. PMID- 14526839 TI - Tuning of a Ti3+:sapphire laser by an electro-optic beam deflection method. AB - A wavelength-switching method for tuning a self-injection-seeded Ti3+:sapphire laser that uses an electro-optic beam deflection technique is reported. A LiNbO3 prism was employed in a tuning arm of the dual-cavity Ti3+:sapphire laser, and wavelength tuning of approximately 94 pm was attained by altering the deflection angle with the application of an electric field of 10 kV/cm to the prism. The spectral characteristics of the output laser were mainly determined by the diffraction grating in the dual-cavity laser, and the electro-optic prism just behaved as a light-beam deflector for the wavelength tuning purpose. This configuration can allow a simple tuning approach where fast and stable electronic wavelength switching is required in a narrow tuning range, on an order between a few tens of picometers to nanometers, without involving any mechanical movement. PMID- 14526840 TI - Doppler-induced unidirectional operation of a continuous-wave Raman ring laser in H2. AB - We demonstrate stable operation of a diode-pumped cw Raman ring laser in diatomic hydrogen gas. Doppler-induced asymmetry between the the forward and the backward Raman gains leads to inherent unidirectional operation in the forward direction without intracavity optical elements. Use of the ring-cavity geometry dilutes the deleterious effects of thermal lensing and significantly reduces optical feedback to the pump laser. PMID- 14526842 TI - Wideband numerical modeling for vertical-cavity surface-emitting laser Bragg reflectors. AB - We present a novel, to our knowledge, scheme for vertical-cavity surface-emitting laser Bragg reflectors modeling in wideband systems, which notably improves accuracy and numerical efficiency. These Bragg reflectors are characterized as digital filters and then are approximated by use of autoregressive moving-average models. The features of the models are analyzed in terms of the predicted error in the frequency domain, keeping it below 0.1% over a 85-nm range. The numerical structure allows the Bragg reflectors to be integrated accurately into dynamic models of tunable lasers and amplifiers. PMID- 14526841 TI - Wavelength-tunable optical short-pulse generation by mutual pulse injection seeding of two gain-switched Fabry-Perot laser diodes. AB - A simple and robust system is presented to generate wavelength-tunable optical short pulses by use of two gain-switched Fabry-Perot laser diodes in a mutual pulse injection-seeding scheme. The operating wavelength of the optical pulses is flexibly selected by adjustment of a tunable filter, and its intensity is enhanced with an erbium-doped fiber amplifier. The side-mode suppression ratio achieved by the system is larger than 26 dB over a wavelength region of 25 nm and higher than 31 dB within an 18-nm wavelength. PMID- 14526845 TI - Morphology-dependent resonances of nearly spherical particles in random orientation. AB - We use precise T-matrix calculations for prolate and oblate spheroids, Chebyshev particles, and spheres cut by a plane to study the evolution of Lorenz-Mie morphology-dependent resonances (MDRs) with increasing asphericity of nearly spherical particles in random orientation. We show that, in the case of spheroids and Chebyshev particles, the deformation of a sphere by as little as one hundredth of a wavelength essentially annihilates supernarrow MDRs, whereas significantly larger asphericities are needed to suppress broader resonance features. The MDR position and profile are also affected when the deviation of the particle shape is increased from that of a perfect sphere. In the case of a sphere cut by a plane, the supernarrow MDRs are much more resistant to an increase in asphericity and do not change their position and profile. These findings are consistent with the widely accepted physical interpretation of the Lorenz-Mie MDRs. PMID- 14526844 TI - Effect of surface degradation on optical performance of potassium dihydrogen phosphate optics. AB - We have measured scatter produced by roughening of bare potassium dihydrogen phosphate (KDP) surfaces and by surface degradation (etch pits) that develop beneath a porous solgel coating on KDP after exposure to ambient relative humidity. The etch pits that form on coated KDP scatter incident light into strongly anisotropic angular distributions characteristic of the defect size and shape. The total integrated scatter (TIS) can be as high as 9% for a crystal with etch pits as compared with 0.05% for the as-manufactured crystal. The amount of TIS correlates with the area obscured by defects as measured by optical microscopy. PMID- 14526843 TI - Surface aerosol radiative forcing derived from collocated ground-based radiometric observations during PRIDE, SAFARI, and ACE-Asia. AB - An approach is presented to estimate the surface aerosol radiative forcing by use of collocated cloud-screened narrowband spectral and thermal-offset-corrected radiometric observations during the Puerto Rico Dust Experiment 2000, South African Fire Atmosphere Research Initiative (SAFARI) 2000, and Aerosol Characterization Experiment-Asia 2001. We show that aerosol optical depths from the Multiple-Filter Rotating Shadowband Radiometer data match closely with those from the Cimel sunphotometer data for two SAFARI-2000 dates. The observed aerosol radiative forcings were interpreted on the basis of results from the Fu-Liou radiative transfer model, and, in some cases, cross checked with satellite derived forcing parameters. Values of the aerosol radiative forcing and forcing efficiency, which quantifies the sensitivity of the surface fluxes to the aerosol optical depth, were generated on the basis of a differential technique for all three campaigns, and their scientific significance is discussed. PMID- 14526846 TI - Polarized infrared emissivity from dew droplets on a surface. AB - We model the infrared polarization emissivity from spherical particles on a plane surface. The emissivity and polarization is primarily a function of the density of particles multiplied by their cross-sectional area. The presence of particles tends to reduce the polarization. As the emission angle increases from near normal incidence, the polarization tends to pass through a maximum, followed by a minimum and a final sharp rise at near grazing angles. The mechanism for this structure is the shadowing of different portions of the spherical particles by other particles. PMID- 14526847 TI - Internally excited acoustic resonator for photoacoustic trace detection. AB - The quantum-cascade laser can be used as an infrared source for a small portable photoacoustic trace gas detector. The device that we describe uses a quantum cascade laser without collimating optics mounted inside an acoustic resonator. The laser is positioned in the center of a longitudinal resonator at a pressure antinode and emits radiation along the length of the resonator exciting an axially symmetric longitudinal acoustic mode of an open-ended cylindrical resonator. Experiments are reported with an 8-microm, quasi-cw-modulated, room temperature laser used to detect N2O. PMID- 14526848 TI - Excitation saturation in two-photon fluorescence correlation spectroscopy. AB - Fluorescence correlation spectroscopy (FCS) has become a powerful and sensitive research tool for the study of molecular dynamics at the single-molecule level. Because photophysical dynamics often dramatically influence FCS measurements, the role of various photophysical processes in FCS measurements must be understood to accurately interpret FCS data. We describe the role of excitation saturation in two-photon fluorescence correlation measurements. We introduce a physical model that characterizes the effects of excitation saturation on the size and shape of the two-photon fluorescence observation volume and derive a new analytical expression for fluorescence correlation functions that includes the influence of saturation. With this model, we can accurately describe both the temporal decay and the amplitude of measured fluorescence correlation functions over a wide range of illumination powers. PMID- 14526849 TI - Time-resolved laser-induced incandescence of soot: the influence of experimental factors and microphysical mechanisms. AB - We present a data set for testing models of time-resolved laser-induced incandescence of soot. Measurements were made in a laminar ethene diffusion flame over a wide range of laser fluences at 532 nm. The laser was seeded to provide a smooth temporal profile, and the beam was spatially filtered and imaged into the flame to provide a homogeneous spatial profile. The particle incandescence was imaged onto a fast photodiode. The measurements are compared with the standard Melton model [Appl. Opt. 23,2201 (1984)] and with a new model that incorporates physical mechanisms not included in the Melton model. PMID- 14526850 TI - Nonlinear refraction and saturable absorption in Au:BaTiO3 composite films. AB - Composite thin films Au:BaTiO3, comprising nanometer-sized gold particles embedded in barium titanate matrices, were synthesized on MgO (100) substrates with the pulsed laser deposition technique. The nanostructure of the films and the size distributions of the Au particles were analyzed by high-resolution transmission electron microscopy. Crystal lattice fringes from the Au nanocrystals and the BaTiO3 matrices were observed. The nonlinear optical properties of the Au:BaTiO3 films were measured with the z-scan method at a wavelength of 532 nm, which was closed to the surface plasmon resonance of nanoscale Au particles. The features of the closed-aperture z-scan transmittance curves were affected by the ratio, which increased greatly at a high metal concentration, of the imaginary part to the real part of the third-order nonlinear susceptibility chi(3). PMID- 14526851 TI - Femtosecond optical parametric generation of noncollinear phase matching for a biaxial crystal. AB - In an optical parametric generation of a femtosecond pulse for a biaxial crystal, the interaction of three waves can be used as a model of noncollinear phase matching in which the group velocities of the interacting pulses are suitably linked to each other. For satisfaction of group-velocity matching, the tunable parametric generation of femtosecond pulses must use noncollinear phase matching. We consider three conditions of group-velocity matching for femtosecond pulses. Signal and idler pulses can be obtained when the coupled-wave equations, including the group-velocity mismatch and group-velocity dispersion effects, are solved. A Fourier method is an effective method for solving the equations, and from the solution of the equations the relation between duration of output pulses and wavelengths can be obtained. In a comparison of collinear and noncollinear matches, when the latter is group-velocity matched, the duration of its outpulses are smaller, and the outpulses can be continually tuned from the visible to the mid-infrared. PMID- 14526852 TI - What is residency? PMID- 14526853 TI - Rethinking the way we manage medications. Using pharmacists in community family practice. PMID- 14526854 TI - Weight loss, not drugs. PMID- 14526855 TI - Learning the lessons of the HRT fiasco. PMID- 14526856 TI - Influencing public policy. PMID- 14526858 TI - Trimethoprim-sulfonamide combination therapy in early pregnancy. AB - QUESTION: One of my patients presented with bacteriuria early in her pregnancy. Urine culture was positive for Escherichia coli. I would like to prescribe a trimethoprim-sulfamethoxazole combination because it worked well for her in the past. What is known about the safety of this medication during early pregnancy? ANSWER: Evidence-based studies report an association between trimethoprim sulfonamide combinations in early pregnancy and several major malformations, such as neural tube defects and cardiovascular defects. If clinically possible, physicians are advised to use alternative antimicrobial medications for treatment of urinary tract infections during early pregnancy. PMID- 14526857 TI - Returning to residency. PMID- 14526859 TI - Dermacase. Scleredema adultorum, or Bushke disease. PMID- 14526861 TI - Circumcision and cervical cancer. Is there a link? PMID- 14526860 TI - Practice tips. Real-time digital photography. Adjunct to medical consultation. PMID- 14526862 TI - Preventing running injuries. Practical approach for family doctors. AB - OBJECTIVE: To present a practical approach for preventing running injuries. QUALITY OF EVIDENCE: Much of the research on running injuries is in the form of expert opinion and comparison trials. Recent systematic reviews have summarized research in orthotics, stretching before running, and interventions to prevent soft tissue injuries. MAIN MESSAGE: The most common factors implicated in running injuries are errors in training methods, inappropriate training surfaces and running shoes, malalignment of the leg, and muscle weakness and inflexibility. Runners can reduce risk of injury by using established training programs that gradually increase distance or time of running and provide appropriate rest. Orthoses and heel lifts can correct malalignments of the leg. Running shoes appropriate for runners' foot types should be selected. Lower-extremity strength and flexibility programs should be added to training. Select appropriate surfaces for training and introduce changes gradually. CONCLUSION: Prevention addresses factors proven to cause running injuries. Unfortunately, injury is often the first sign of fault in running programs, so patients should be taught to recognize early symptoms of injury. PMID- 14526863 TI - Paying more than lip service to lip lesions. AB - OBJECTIVE: To review the epidemiology, etiology, diagnosis, management, and prognosis of the most common, potentially lethal, lip lesions: leukoplakia, actinic cheilitis, and squamous cell carcinoma (SCC). QUALITY OF EVIDENCE MEDLINE: was searched from 1966 to 2002 for English-language articles on prevalence of lip lesions. No articles for a family physician audience were found. MEDLINE was searched again using the terms "leukoplakia," "actinic cheilitis," and "squamous cell carcinoma." Randomized, controlled trials were selected; non-blinded trials, population-based studies, and systematic reviews were also used. MAIN MESSAGE: Leukoplakia, actinic cheilitis, and SCC of the lips are relatively common presentations that can cause substantial morbidity and, more rarely, mortality. Any abnormality of the lips can be an embarrassment. Because of the seriousness and frequency of lip disease, it is important to look for, diagnose, and treat lip lesions to prevent morbidity and mortality and also to maintain social acceptance and self-esteem. CONCLUSION: Knowledge of leukoplakia, actinic cheilitis, and SCC of the lips will aid family physicians in diagnosing and managing these lesions and in preventing associated morbidity and mortality. PMID- 14526864 TI - Physician do not heal thyself. Survey of personal health practices among medical residents. AB - OBJECTIVE: To assess how many residents follow the recommendation that physicians have a personal family physician and where residents seek medical attention when needed. DESIGN: Hand-delivered survey. SETTING Residency training programs at Queen's University. PARTICIPANTS: Of 215 residents with a central mailbox, 122 responded (response rate 57%). MAIN OUTCOME MEASURES: Health status, usual access to health care, having a personal family physician, and response to two scenarios. RESULTS: More than a third (38%) of residents have a local family physician, yet 25% of those with chronic illnesses and 40% of those who use prescription medications regularly do not. Many rely on colleagues; 41% have received prescriptions from or written prescriptions for their colleagues. Residents with local family physicians are more likely to seek appropriate medical attention for physical problems. Residents do not recognize or seek treatment for mental health problems. Knowledge, time, and accessibility were considered barriers to adequate health care. CONCLUSION: Many residents do not have good access to comprehensive, confidential, and objective medical care. They rely on colleagues, and they ignore mental health problems. Lack of time and access, and attitudes about the importance of having a family physician are important barriers. PMID- 14526865 TI - Choosing family medicine. What influences medical students? AB - OBJECTIVE: To explore factors that influence senior medical students to pursue careers in family medicine. DESIGN: Qualitative study using semistructured interviews. SETTING: University of Western Ontario (UWO) in London. PARTICIPANTS: Eleven of 29 graduating UWO medical students matched to Canadian family medicine residency programs beginning in July 2001. METHOD: Eleven semistructured interviews were conducted with a maximum variation sample of medical students. Interviews were transcribed and reviewed independently, and a constant comparative approach was used by the team to analyze the data. MAIN FINDINGS: Family physician mentors were an important influence on participants' decisions to pursue careers in family medicine. Participants followed one of three pathways to selecting family medicine: from an early decision to pursue family medicine, from initial uncertainty about career choice, or from an early decision to specialize and a change of mind. CONCLUSION: The perception of a wide scope of practice attracts candidates to family medicine. Having more family medicine role models early in medical school might encourage more medical students to select careers in family medicine. PMID- 14526868 TI - Improving competence in maternity care. Women's Advanced Maternity Fellowship for Rural Practitioners. PMID- 14526867 TI - Clinical calculators for hand-held computers. PMID- 14526866 TI - Keeping family physicians in rural practice. Solutions favoured by rural physicians and family medicine residents. AB - OBJECTIVE: To determine how family medicine residents and practising rural physicians rate possible solutions for recruiting and sustaining physicians in rural practice. DESIGN: Cross-sectional mailed survey. SETTING: Rural family practices and family medicine residency programs in Ontario. PARTICIPANTS: Two hundred seventy-six physicians and 210 residents. MAIN OUTCOME MEASURES Ratings of proposed solutions on a 4-point scale from "very unimportant" to "very important". RESULTS: Rural family physicians rated funding for learner-driven continuing medical education (CME) and limiting on-call duty to 1 night in 5 as the most important education and practice solutions, respectively. Residents rated an alternate payment plan to include time off for attending and teaching CME and comprehensive payment plans with a guaranteed income for locums as the most important education and practice solutions, respectively. CONCLUSION: Residents and physicians rated solutions very similarly. A comprehensive package of the highest-rated solutions could help recruit and sustain physicians in rural practice because the solutions were developed by experts on rural practice and rated by family medicine residents and practising rural physicians. PMID- 14526870 TI - Measuring psychological stress. Concept, model, and measurement instrument in primary care research. PMID- 14526871 TI - Locum tenens. Having a win-win experience. PMID- 14526869 TI - Relishing the moment. Having a baby in residency. PMID- 14526872 TI - Family medicine's future: in our hands. PMID- 14526873 TI - The future of health social work. AB - The practice of social work in health care is at a critical juncture, and laces an uncertain future. The authors provide an overview of the challenges facing social work practice within the health care setting, as well as recommendations for enhancing social work practice and education. Challenges discussed include economic factors, demographic changes, and technological advances influencing the practice of social work in health care. The need for a proactive stance among social work professionals and educators is promoted. The proposed changes are intended to stimulate discussion and an exchange of ideas needed to maintain Social Work's relevance and integrity in the evolving health care delivery system. PMID- 14526875 TI - Are they grieving? A qualitative analysis examining grief in caregivers of individuals with Alzheimer's disease. AB - Although there is an abundance of caregiving literature, there is a paucity of research that examines the grief reactions of caregivers of individuals with Alzheimer's disease. Qualitative research methodology was utilized to identify themes about the grief reactions expressed by caregivers. Results found that although some caregivers reported that they were not grieving, there was still a strong sense of loss in their descriptions of their caregiving experience. Additionally, for the caregivers who reported that they were grieving, their grief was characterized by multiple losses that accumulated during the disease process. PMID- 14526874 TI - Effect of gender on the social and psychological adjustment of cancer patients. AB - The purpose of this article is to examine gender differences in social and psychological adjustment among cancer patients. The social adjustment and psychological distress of 49 patients (34 women and 15 men) undergoing active medical care (chemotherapy and radiation) were assessed. Socio-demographic and medical parameters were also examined to account for differences in adjustment. Three questionnaires were used: A personal information questionnaire; a Psychosocial Adjustment to Illness Scale (PAIS-SR) (Derogatis & Lopez, 1983); and a psychological distress one (BSI) (Derogatis & Spencer, 1982). Significant differences were found between men and women patients in several dimensions of the social adjustment, psychological distress and medical variables. Possible explanations, recommendations for further research and clinical intervention are suggested. PMID- 14526876 TI - Psychological adaptation and distress among HIV+ Latina women: adaptation to HIV in a Mexican American cultural context. AB - In order to examine the relative importance of general individual orientations (mastery and self-esteem) and specific coping styles with regard to psychological distress among women with HIV, data are examined from a pilot study involving in depth face-to-face interviews with 32 HIV+ Latinas receiving care at 4 clinics serving the South Texas population. Interviews were conducted to provide preliminary psychometric information on coping and distress instruments in this predominantly Mexican American population as well as to examine psychosocial factors related to individual adjustment to HIV among Latina females. Refusal rates were low in this study (approximately 10%) and measurement instruments generally had acceptable internal consistency reliability. Results of exploratory multiple regression analyses suggest that self-esteem and mastery may be more salient predictors of depression and anxiety symptoms than are specific coping strategies. PMID- 14526878 TI - European Society of Medical Oncology (ESMO) joins the palliative care community. PMID- 14526877 TI - Achieving equity in referrals for renal transplant evaluations with African American patients: the role of nephrology social workers. AB - Research over the last 12 years has repeatedly shown that African-American ESRD patients are less likely than white patients to be referred for a transplant evaluation. This article takes an exploratory look at issues surrounding this racial disparity. Among the issues focused on are: (a) the multiple ramifications from this racial disparity; (b) neglected factors which may be militating against the equitable referral of African-American patients; and (c) the vulnerabilities of this patient population. The final focus is on intervention strategies for nephrology social workers and how they might help to facilitate a greater equity in this area of health care with African-American patients. PMID- 14526879 TI - High prevalence of non-albicans yeasts and detection of anti-fungal resistance in the oral flora of patients with advanced cancer. AB - Oral fungal infections frequently develop in individuals with advanced cancer. This study examined the oral mycological flora of 207 patients receiving palliative care for advanced malignant disease. Demographic details and a clinical history were documented from each participant. A tongue swab was collected and cultured on CHROMAgar Candida (CHROMAgar Paris, France). All yeasts were identified by germ tube test, API ID 32C profiles and, for Candida dubliniensis, by species-specific PCR. Susceptibility to fluconazole and itraconazole was determined by a broth microdilution assay according to the National Committee for Clinical Laboratory Standards (NCCLS). At time of sampling, 54 (26%) of the 207 subjects had clinical evidence of a fungal infection and yeasts were isolated from 139 (67%) individuals. In total, 194 yeasts were isolated, of which 95 (49%) were Candida albicans. There was a high prevalence of Candidia glabrata (47 isolates) of which 34 (72%) were resistant to both fluconazole and itraconazole. All nine isolates of C. dubliniensis recovered were susceptible to both azoles. No relationship was established between anti fungal usage in the preceding three months and the presence of azole resistant yeasts. This study of patients with advanced cancer has demonstrated a high incidence of oral colonization with non-C. albicans yeasts, many of which had reduced susceptibility to fluconazole and itraconazole. The role of improved oral care regimes and novel anti-fungal drugs merits further attention, to reduce the occurrence of fungal infection in these patients. PMID- 14526881 TI - Attitudes of patients and staff to research in a specialist palliative care unit. AB - Many randomized trials in specialist palliative care (SPC) have failed to recruit sufficient numbers of patients. A questionnaire was designed to assess the attitudes of inpatients of a SPC unit to taking part in research. Only 50% of patients were considered physically and mentally fit for interview. Forty patients and 13 nurses were asked to indicate their willingness to take part, or for their patients to take part, in research in general and then in three hypothetical trials. Reasons for their responses were analysed. The patients were generally agreeable to trial participation (66% 'quite happy' or 'very happy'). The nurses were strongly in favour of research participation for the same group of patients (92% 'quite happy' or 'very happy'). The most invasive study involving venepuncture and random drug allocation had the least favourable response (46% of patients and 54% of nurses 'quite happy' or 'very happy'). A trial of reflexology attracted 77% of patients, while 62% were happy to take part in a study involving mood assessment and interview. Factors deterring willingness to participate included the need for record keeping by the patient and concern about potential emotional strain. The nurses correctly identified some of the factors deterring patients, but often their willingness for trial participation did not match that of the patient. Although there is support for research among the small number of potential trial entrants in specialist palliative care units, their limited physical and emotional reserves make careful attention to appropriate trial design essential to the success of future studies. PMID- 14526880 TI - Palliative care research protocols: a special case for ethical review? AB - Between October 2001 and May 2002 the Chairperson and Vice-Chairperson of each Multicentre Research Ethics Committee (MREC) in England, Wales and Scotland took part in a semi-structured interview to ascertain the attitudes of MRECs to palliative care research. Interviews were transcribed and analysed using a grounded theory approach. Most respondents said each protocol was reviewed on its own merits, according to broad ethical principles, but were equivocal as to whether palliative care protocols posed particular or different challenges compared to those from other specialties. Respondents said they reviewed only a small number of palliative care protocols, and that they were less experienced with some of the study methods utilized, particularly qualitative designs. Four main themes emerged from the analysis. Respondents expressed concerns about the protocol itself--in regard to safeguarding the principles of autonomy and justice. There were concerns about how the research would be carried out, especially the protection of patients and the influence and input of the researcher in the process. The third theme concerned the impact of the research on the participant, particularly intrusion, potential distress and the existence of support mechanisms. Fourthly, respondents identified patient groups receiving palliative care (children, the elderly, bereaved families, patients in intensive therapy units, and those from ethnic groupings), who they considered might be particularly vulnerable. PMID- 14526883 TI - Symptoms in advanced cancer: relationship to endogenous cortisol levels. AB - We investigated a possible relationship between levels of endogenous cortisol and severity of different symptoms in patients with advanced cancer. Twenty-three patients with predominantly gastrointestinal cancer, recruited in a palliative care unit, entered the study. Urinary free cortisol (UFC) was measured together with demographic data, blood parameters, tumour burden, concurrent illness, medication, nutritional status and quality of life. Significant positive correlations were found between levels of endogenous cortisol and appetite loss, fatigue and nausea/vomiting. The findings support the view of a chronic stress condition in advanced cancer. Interaction between cytokines and the hypothalamic pituitary-adrenal (HPA) axis may also be important in the interpretation of the results. PMID- 14526882 TI - Hospital based palliative care teams improve the symptoms of cancer patients. AB - Despite the increase in hospital palliative care teams, there is little research into their impact on symptom control in patients. A nonequivalent control group design, using a quota sample, investigated 100 cancer patients who had been admitted to hospital for symptom control. Fifty patients received hospital palliative care team intervention compared with 50 patients receiving traditional care. Data was collected using the Palliative Care Assessment (PACA) symptom assessment tool on three occasions. Both groups showed a statistically significant improvement in their symptoms. This significance failed to meet the criterion of one point on a four point scale and therefore results have to be interpreted with caution. However the intervention group had a greater improvement in all their symptoms, particularly for the pain and anorexia for which there were no differences between the groups on the initial assessment, there was a statistically greater improvement for the intervention group (P < 0.001). Consecutive patients with cancer admitted to hospital for symptom control during this study improved, but those patients who received specialist palliative care had a significantly greater improvement in their symptoms. PMID- 14526884 TI - Assessing the independent contribution to quality of life from anxiety and depression in patients with advanced cancer. AB - BACKGROUND: The aim of palliative care services is to ensure the best quality of life for patients and their carers. Depression is common amongst palliative care patients and has been shown to adversely affect quality of life. This study aimed to examine the independent contribution of depression to quality of life. OBJECTIVE: To investigate the hypothesis that a) illness severity, pain, anxiety and depression are all associated with impaired health-related quality of life and b) once the effects of illness severity have been adjusted for, there is further independent contribution to quality of life from anxiety and depression. METHOD: Consecutive patients with advanced cancer under the care of palliative care services were screened. Sixty-eight patients were evaluated for levels of anxiety and depression, severity of illness, pain severity and health-related quality of life. RESULTS: Thirty-three males and 35 females were recruited and had an age range of 41-93 years (median 71). Seventeen (25%) of patients were anxious [anxiety score > or = 11 on the Hospital Anxiety and Depression Scale (HADS)], 15 (22%) were depressed (HADS depression score > or = 11). After controlling for the effects of pain and illness severity, anxiety and depression were independently associated with global health status, emotional and cognitive functioning, and fatigue. Anxiety further contributed significantly towards social functioning, nausea and vomiting. CONCLUSIONS: This study has confirmed that pain, anxiety and depression were associated with impaired quality of life. Anxiety and depression contributed independently towards various dimensions of quality of life. Longitudinal studies are required to examine the direction of the causal association between pain and depression in patients receiving palliative care. PMID- 14526885 TI - Palliative care for patients with end-stage renal failure: reflections from Central Australia. AB - This paper explores palliative care issues for Australian Aboriginal patients with end-stage renal failure. Renal disease is epidemic amongst Aboriginal Australians. The Central Australian Palliative Care Service, based in the remote town of Alice Springs, sees a high proportion of renal patients, with different demographic characteristics to those seen in urban palliative care services. A case history illustrates distinctive concerns for local service provision, including communication issues, cultural requirements for maximizing quality of life, and the need to provide care in remote locations. Palliation for end-stage renal failure (ESRF) patients should be based on standard principles, but modified in accordance with local practical requirements and community needs. PMID- 14526889 TI - Oral medicine. PMID- 14526886 TI - Difficulties encountered by nurses in the care of terminally ill cancer patients in general hospitals in Japan. AB - The aim of this study was to investigate the difficulties encountered by nurses who have cared for terminally ill cancer patients at general hospitals. To collect data, a survey by questionnaire was self-administered. The respondents were 375 nurses and the response rate was 70.2%. Factor analysis was conducted on 80 items related to the difficulties encountered by nurses who have cared for terminally ill cancer patients to allow reasonable item reduction and to explore better domains. Two items were excluded and the results revealed eight underlying domains: 'Communication with patients and families', 'Knowledge and skill of nurses', 'Treatment and informed consent (IC)', 'Personal issues', 'Collaboration as a team including patients and families', 'Environment and system', 'Collaboration among nurses', and 'Near-death issues'. Cronbach's alpha coefficients for each domain ranged from 0.77 to 0.93. The results showed that nurses working at general hospitals have experienced a high degree of difficulty overall while caring for the dying, particularly with communication with patients and families. It was concluded that this study was useful in determining the specific areas where nursing education and research should be focused. PMID- 14526887 TI - Psychosocial morbidity associated with patterns of family functioning in palliative care: baseline data from the Family Focused Grief Therapy controlled trial. AB - Family Focused Grief Therapy (FFGT) is a new model of brief intervention, which is commenced during palliative care for those families shown to be at high risk of poor adaptation, and continued preventively into bereavement with the aim of improving family functioning and reducing the morbid consequences of grief. In this paper, baseline data on 81 families (363 individuals) selected by screening from a palliative care population are explored to confirm our previously reported observation that high levels of psychosocial morbidity are positively associated with worsening family dysfunction. The Family Relationships Index (FRI) was used for screening and the Family Assessment Device (FAD) as an independent family outcome measure. The Beck Depression Inventory (BDI), Brief Symptom Inventory (BSI) and Social Adjustment Scale (SAS) were the psychosocial measures. Families were classified according to their functioning based on the FRI. To allow for correlated family data, statistical analyses employed the generalized estimating equation (GEE) method, controlling for gender and depression (BDI). Screening of 257 families (701 individuals) revealed 74 (29%) well-functioning families and 183 (71%) at some risk of morbid outcome. Of the latter, 81 (44%) gave informed consent to enter a randomized controlled trial of FFGT. Patients had a mean age of 57 years, 51% were male and they suffered from cancer, with a median length of illness from diagnosis to death of 25 months. In accordance with the FFGT model, their family types were identified as Intermediate 51%, Sullen 26% and Hostile 23%. These were significantly associated with steadily increasing levels of distress (BSI) and poor social adjustment (SAS). The FAD confirmed the concurrent accuracy of the FRI. As significantly greater levels of psychosocial morbidity were present in families whose functioning as a group was poorer, support was generated for a clinical approach that screens for families rather than individuals at high risk. The predictive validity of the FRI as a screening measure was confirmed. Overall, these baseline data point to the importance of a family-centred model of care. PMID- 14526888 TI - The use of zoledronic acid in the management of metastatic bone disease and hypercalcaemia. AB - Zoledronic acid is a potent, third generation, nitrogen-containing bisphosphonate, licensed for the management of skeletal metastases and hypercalcaemia of malignancy, both of which cause considerable morbidity. In the preclinical setting, zoledronic acid has demonstrated superior potency regarding inhibition of osteolysis and reduction of hypercalcaemia as compared with other bisphosphonates. Clinical trials have indicated that zoledronic acid is superior to pamidronate in suppressing osteolysis and in reducing hypercalcaemia of malignancy. Its main mechanism of action is induction of osteoclast apoptosis through inhibition of the mevalonate pathway. Zoledronic acid has also demonstrated direct anti-tumour activity both in vitro and in animal models, suggesting it may be of benefit in preventing the formation of bone metastases. Clinical trials are in progress, assessing the benefit of zoledronic acid in the adjuvant setting in both breast and prostate cancer. PMID- 14526890 TI - Strategies for recruiting patients into randomized trials of palliative care. PMID- 14526891 TI - A trial that failed because of poor recruitment. PMID- 14526892 TI - How do patients with primary brain tumours die? PMID- 14526894 TI - Single or shared rooms for hospice patients? PMID- 14526893 TI - Swallowing capsules. PMID- 14526895 TI - Allied health legislation redux. PMID- 14526897 TI - The relationship between self-efficacy and student physician assistant clinical performance. AB - The purpose of this study was to investigate the predictive relationship between student self-efficacy beliefs, achievement expectations, perceived outcome values, and subsequent clinical performance. The study investigated whether measures of self-efficacy, measures of achievement expectancy, or measures of perceived task value (relevance) correlated significantly with physician assistant clinical performance; also, it investigated whether utilizing both noncognitive criteria and cognitive criteria will improve upon the predictability of physician assistant clinical performance. The independent variables include grade point average, previous health care experience, self-efficacy, values, and outcome expectations. The research population comprised a group of approximately 300 students attending the 14 accredited programs within Pennsylvania who, at the time of the surveys, were beginning the clinical year of their professional education. Data collection methods included surveys and quantitative data sources. The data were subjected to multivariate statistical methods including multiple regression and discriminate analysis. The analysis included both correlation and regression to predict clinical performance from the noncognitive and cognitive variables. The results reveal that self-efficacy, and not just academic performance, is a significant predictor of a student's clinical performance. Utilizing both noncognitive and cognitive variables can improve upon the predictability of the student physician assistant's clinical performance. PMID- 14526896 TI - Correlates of work exhaustion for medical technologists. AB - In a sample of 196 medical technologists followed over a 4-year period, this study investigated if work-related demand and resource variables were related to subsequent work exhaustion. As hypothesized, increased levels of perceived work interference with family and task load and lower organizational support were related to higher subsequent work exhaustion. Distributive justice, as an intervening variable, had direct and partially mediating effects on work exhaustion. Distributive justice partially mediated the effects of work interfering with family and organizational support on work exhaustion. Distributive justice also mediated the impact of procedural justice on work exhaustion. Study limitations and future research issues are discussed. PMID- 14526898 TI - Elements of instruction in allied health: do faculty and students value the same things? AB - The purposes of this study were to determine the importance placed on 15 elements of instruction in allied health by students and faculty and to identify similarities and differences in students' and faculty's views. A total of 145 students and 55 faculty in the College of Allied Health at the University of Oklahoma participated in an on-line survey. Focus groups subsequently were conducted to discuss areas of agreement and disagreement between students and faculty. Four common elements were contained in the top-five list of both groups: (1) the teacher's ability to relate course material to clinical situations, (2) the teacher's communication skills, (3) the teacher's preparation and organization, and (4) the teacher's knowledge of the subject. Five of the 15 items were significantly different: (1) the teachers' friendliness and respect for students, (2) the teacher's ability to motivate students, (3) the teacher's ability to challenge students intellectually, (4) the teacher's encouragement of discussion, and (5) the teacher's ability to use a number of teaching techniques. Results of this study will further understanding between allied health faculty and students concerning similarities and differences in instructional values. PMID- 14526899 TI - Patterns of safer sex practices among allied health students at historically black colleges and universities. AB - With high human immunodeficiency virus (HIV) and STD transmission rates among African American young adults, there is a need to study the patterns of risk behaviors among this group for effective HIV prevention strategies. Many studies point to the problem of what is termed the knowledge-behavior gap, as a contributing factor for high transmission rates. In this phenomenon, the risks for HIV transmission are known, yet this knowledge does not translate into safer sex practices. It is unclear if this phenomenon applies to all young adults. This study examined sexual behaviors, risks for HIV infection, condom use, and HIV testing practices among allied health students. A sample of 614 respondents voluntarily completed questionnaires. The respondents were from seven historically black colleges and universities and four predominantly African American physician assistant programs. The significant findings reflect that this sample of mature and informed allied health students engaged in safer sex practices. There was a higher rate of condom use among African Americans compared with other racial and ethnic groups and significant differences for gender, race, and antibody testing. The outcomes of the study have implications for using allied health students as role models in designing effective prevention programs on college campuses and in African American communities to address knowledge behavior gap issues. PMID- 14526900 TI - Model for development of outcome assessment surveys for allied health educational programs. AB - This article describes a model for survey development used by the Department of Physical Therapy at Saint Louis University to assess the extent to which the program prepares students for a career as physical therapists. From a review of departmental documents including mission and philosophy statements and curriculum goals, eight performance outcomes (e.g., competence as a general practitioner, leadership, ethics, attitude for service, etc.) and two program characteristics (curricuum and program atmosphere) were determined. External documents including accreditation criteria for graduate performance were also reviewed and found to be consistent with the identified characteristics. Based on these findings, items for three surveys were developed and assessed. A 1-year alumni survey solicited feedback on the graduates' perceptions of their educational preparedness for clinical practice. The graduates' employers then were surveyed for an assessment of the graduates' job performance. A 3-year alumni survey followed up to reassess the graduates' perceptions of their educational preparedness and to track the alumni's activities since graduation. Other programs in physical therapy and other health care disciplines can use this model to develop valid survey instruments to assess their program's effectiveness. PMID- 14526902 TI - Cultural competence of faculty and students in a school of allied health. AB - The School of Allied Health Sciences (SAHS) at East Carolina University administered a cultural competence survey to assess baseline competence for comparison with future, longitudinal assessments. This cultural competence assessment of students and faculty offers a comparison groupfor other schools of allied health that conduct similar studies. The faculty response rate was 59% with 35 completed surveys, and the student response rate was 28% with 151 respondents. The SAHS response rate for the faculty and students combined was 32% (N = 186). The allied health students consistently had higher mean scores than the faculty on knowledge of communities, personal involvement, and total score, although the differences were not statistically significant. This difference may be a reflection of the greater, although still relatively low, ethnic/cultural diversity among the students (82% white) compared with the faculty (97% white). The results are limited in generalizability to other schools of allied health with a predominately white female population. PMID- 14526901 TI - Copyright in the digital classroom. AB - Copyright law was developed to protect the rights of authors. Many educators are aware of copyright law, but they may not be aware of how it applies to them or of the law's scope in regards to computer technology. As the computer is used increasingly to disseminate information, teaching professionals also must have knowledge of the applications of the law to this developing technology. This article reviews the laws defining copyright and fair use. It examines case law regarding copyright, computers, and the Internet. Recommendations for using computers to disseminate class materials to students are discussed. PMID- 14526906 TI - Dichotenone-a and -b: two new enones from the marine brown alga Dictyota dichotoma (Hudson) Lamour. AB - Marine brown alga Dictyota dichotoma (Dictyotaceae) collected from Karachi coast of Arabian Sea yielded two new (1 and 2) enones (dolastane-diterpenoids) named: dichotenone-A and -B along with an olide (loliolide, 3) as a new source. Their structures have been characterized with the aid of 2D-NMR spectroscopic techniques. PMID- 14526903 TI - Using memory strategies to improve 24-hour dietary recalls among older adults. AB - Dietitians commonly use 24-hour recall to obtain estimates of the typical food intakes for a population. This study was designed to determine whether using an encoding strategy or support at the time of retrieval improves older adults' 24 hour recall of food items and amounts consumed. The study included 17 older adults ranging from 74 to 91 years of age. Lunch was served to these individuals, and 24-hour dietary recall and recognition of food items consumed at the meal were conducted the following day. Portion sizes for five food items served at the meal, as well as memory tests for recall and recognition and a visual perceptual test, were also administered. Among older adults, use of an encoding strategy did not enhance memory of food items and amounts consumed. However, when the adults were provided support with a visual recognition aid at the time of food item recall, individuals remembered more food items but not amounts consumed. Performance on the Wechsler Memory Scale I and II, Verbal Paired Associates, was found to be a predictor of subjects' performance on 24-hour recall. PMID- 14526904 TI - Physical therapy on trial: the rationale, organization, and impact of a mock trial on physical therapy students' attitudes toward and confidence in research. AB - A learning activity, debate with a mock trial format, provides opportunities for physical therapy students to develop skill and confidence in learning to evaluate research critically, to formulate opinions regarding the credibility of the available evidence for a therapeutic approach to patient care, and to substantiate their choice of treatments with persuasive arguments and analytic rebuttals. The purposes of this study were to (1) describe the mock trial learning activity within a professional physical therapy curriculum and (2) determine whether the mock trial positively influenced students' attitudes toward research and confidence in their ability to interpret and apply research findings to clinical practice. A structured questionnaire was distributed to the 48 students at key points during the academic program to measure the overall effects of the physical therapy curriculum and the specific effects of the mock trial learning activity. There was an increase (p < 0.05) in the students' confidence in their ability to interpret and apply research after participating in the mock trial. The overall responses to the modified Attitudes-to-Research scale indicated that the physical therapy students generally held positive and optimistic attitudes toward research. These attitudes were maintained over the entire experience within the curriculum. If attitudes predict behavior, these future practitioners are more likely to be physical therapists who critically appraise and use evidence to determine practice. PMID- 14526905 TI - Converting a bachelor of health sciences course on communication skills to a CD ROM format. PMID- 14526907 TI - Biopreparation of (-)-(1S,3R,4S,6S)-6-hydroxymenthol and (-)-(1S,3R,4S)-1 hydroxymenthol from 1-menthol by Rhizoctonia solani AG-1-IA and IB. AB - Microbial transformation of 1-menthol (1) by six isolates of soil-borne plant pathogenic fungi Rhizoctonia solani AG-1-IA (Rs24, Joichi-2 and RRG97-1) and AG-1 IB (TR22, R147 and 110.4) as a biocatalyst was investigated. Twenty one days precultivation of Rhizoctonia solani AG-1-IA Rs24 and AG-1-IB 110.4 showed excellent yield (98.5-98.6%) of (-)-(1S,3R,4S,6S)-6-hydroxymenthol (2) and (-) (1S,3R,4S)-1-hydroxymenthol (3) from 1. PMID- 14526910 TI - Two new benzopyran derivatives from Mallotus apelta. AB - From the leaves of Mallotus apelta two new benzopyran compounds (1,2) were obtained and their structures were determined using spectroscopic methods. PMID- 14526911 TI - Albiziahexoside: a potential source of bioactive saponin from the leaves of Albizzia lebbeck. AB - Albiziahexoside (1), a new hexaglycosylated saponin, was isolated from leaves of Albizzia lebbeck. Saponin 1, which is an analog of cytotoxic albiziatrioside A (2), did not show cytotoxicity. However, 1 is a potential source of 2 and related bioactive saponins for medicinal use because leaves, which can be regenerated after collection, contain 1 in substantial quantity. PMID- 14526912 TI - Insecticidal compounds against Drosophila melanogaster from Cornus officinalis Sieb. et Zucc. AB - Dimethyl malate (1) and 5-hydroxymethyl furfural (2) were isolated as insecticidal compounds by bioassay-guided fractionation from MeOH extract of the fruits of Cornus officinalis Sieb. et Zucc. Insecticidal activity against larvae of D. melanogaster was demonstrated: 1 and 2 gave the LC50 value of 6.15 and 11.8 micromol/mL of diet concentration, respectively. Acute toxicity against adults of D. melanogaster, 1 and 2 had the insecticidal activity, with the LD50 value of 21.5 and 34.0 microg/adult. PMID- 14526908 TI - Stereospecific hydroxylation of (+)-cedrol by using human skin microbial flora Staphylococcus epidermidis. AB - Microbial transformation of (+)-cedrol was investigated by using Staphylococcus epidermidis and found that stereospecific hydroxylation of (+)-cedrol occurred at the C-3 position to form (+)-(3S)-3-hydroxycedrol. PMID- 14526914 TI - Substituent effects in the BF3-mediated acylation of phenols with cinnamic acids. AB - 2',4'-Dihydroxy-4-nitrochalcone was obtained by BF3 etherate acylation of resorcinol with 4-nitrocinnamic acid. Instead, when p-methoxycinnamic or p coumaric acids were used, the main product of the reaction was the coumarin umbelliferone, while no chalcone was present. These effects are commented in relation to the nature of the substituent present on the cinnamic acid moiety. PMID- 14526909 TI - Suppressive effect of the SOS-inducing activity of chemical mutagen by citric acid esters from Prunus mume Sieb. Et Zucc. using the Salmonella typhimurium TA1535/pSK1002 umu test. AB - A methanol extract from Prunus mume Sieb. Et Zucc. showed a suppressive effect of the SOS-inducing activity on the mutagen 3-amino-1,4-dimethyl-SH-pyrido[4,3 b]indole(Trp-P-1) in the Salmonella typhimurium TA1535/pSK1002 umu test. The methanol extract was re-extracted with hexane, dichloromethane, ethyl acetate, butanol and water. The dichloromethane and ethyl acetate fractions showed suppressive effect. Suppressive compounds were isolated by silica gel column chromatography and identified as trimethyl citrate (1) and dimethyl citrate (2) by GC-MS, IR and 1H and 13C-NMR spectroscopy. Compounds 1 and 2 suppressed 51 and 39% of the SOS-inducing activity at a concentration of 2.0 micromol/mL. PMID- 14526913 TI - Oviposition-stimulatory activity against Ostrinia zealis by essential oil of root part from Cirsium japonicum DC. AB - The essential oil components from root part of Cirsium japonicum and oviposition stimulatory activity against Ostrinia zealis by root essential oil were investigated. The main component of fresh root essential oil was aplotaxene (75.14%) and of dried root stock was found to be palmitic acid (14.42%). The root oil was significantly more active than the control against O. zealis. PMID- 14526915 TI - Tertiary amines promoted synthesis of symmetrical 1,3-disubstituted ureas. AB - Symmetrical 1,3-disubstituted ureas and cyclic ureas have been synthesized from corresponding isocyanates and diisocyanates by a new versatile, simple and quick method in the presence of tertiary amines at room temperature. The method under investigation has several advantages over the existing techniques, as it is simple to carry out; does not require complicated equipment; work-up is simple, as it does not use expensive chemicals. Moreover, the yields are almost quantitative. It has a potential for commercial applications in pharmaceutical chemistry. PMID- 14526916 TI - Isolation and structure determination of a benzofuran and a bis-nor-isoprenoid from Aspergillus niger grown on the water soluble fraction of Morinda citrifolia Linn. leaves. AB - The leaves of Morinda citrifolia, Linn. afforded a new benzofuran and a bis-nor isoprenoid, blumenol C, hitherto unreported from this source. The structures of these have been elucidated as 5-benzofuran carboxylic acid-6-formyl methyl ester (1) and 4-(3'(R)-hydroxybutyl)-3,5,5, trimethyl-cyclohex-2-en-1-one (2) respectively through spectroscopic studies. The NMR data (including 1D, 2D techniques) and stereochemistry at C-3' of Compound 2 is also being reported for the first time. PMID- 14526917 TI - Isolation and structural elucidation of 5-formyl-2,3-dihydroisocoumarin from Centaurium erythraea aerial parts. AB - An isocoumarin was isolated from the aerial parts of Centauriwn erythraea Rafn. (Gentianaceae). Its structure was established by spectroscopic means as 5-formyl 2,3-dihydroisocoumarin. PMID- 14526921 TI - Radiological protection philosophy for the 21st century. AB - The International Commission on Radiological Protection (ICRP) has stated that its recommendations will be reviewed at least every 10-15 years. It is now some 13 years since the main commission released, for comment, a draft of what was to become the 1990 Recommendations of the ICRP. These have become the basis for international basic safety standards and have been adopted in almost all countries that have radiological protection legislation. Therefore, the ICRP has been stimulating discussion, during the last three years, on the best way of expressing protection philosophy for the next publication of its recommendations, which it hopes will be by 2005. It is now beginning to prepare a draft of these recommendations, with a view to distributing an early version for comment, even though the background work is incomplete. PMID- 14526922 TI - The role of the ICRP in radiation protection--a view from industry. AB - It is the objective of this paper to discuss some aspects concerning the role and importance of the ICRP. Here, this is done with a background of practical radiation protection in industry. The author organises and controls radiation protection for a worldwide operating company, for which efficiently realised radiation safety is as relevant for its workplaces as for its products and services. According to the author's subjective observation, the ICRP has a decreasing importance in operational radiation protection. However, there are growing demands on the ICRP as it is the only basis for internationally compatible regulations and standards. It is the merit of the ICRP that an international comparison of legal protection systems and concepts should give a much more homogeneous picture than that for any other safety and protection issue. The most valuable asset of the ICRP is its credibility as a scientific authority solely committed to the effective protection of people. But its success also brings with it an obligation: there is an increasing need for more effective communication to non-experts. This and other expectations for the future are briefly discussed. PMID- 14526918 TI - A new anthraquinone from Gladiolus gandavensis. AB - A new anthraquinone, 1,6,7-trihydroxy-3-methoxyanthraquinone, along with three known compounds methyl trans-p-hydroxycinnamate, eugenin and 1,3,6-trihydroxy-8 methylanthraquinone, were isolated from the subterranean rhizomes of Gladiolus gandavensis Van Houtt. Their structures were established on the basis of spectroscopic data including 2D NMR techniques and chemical methods. PMID- 14526919 TI - A comparative study of essential oils of Cymbopogon citratus and some members of the genus Citrus. AB - Steam distilled oils of some species of the genus Citrus and Cymbopogon citratus were analyzed by GC-MS. It is observed that citral b was the most common constituent of the oils, which could be a good inhibitor of beta-glucuronidase as all the tested essential oils showed significant inhibition of beta glucuronidase. IC50 values of a mixture of citral a and b also proved the hypothesis. The same oils also exhibited positive response against tested microbes. PMID- 14526920 TI - Evaluation of the antiviral and antimicrobial activities of triterpenes isolated from Euphorbia segetalis. AB - A phytochemical reinvestigation of the whole plant of Euphorbia segetalis yielded five tetracyclic triterpenes: 3beta-hydroxy-cycloart-25-en-24-one (1), cycloart 25-ene-3beta,24-diol (2), cycloart-23-ene-3beta,25-diol (3), lanosta-7,9(11),24 trien-3beta-ol (4) and lanosta-7,9(11),24(31)-trien-3beta-ol (5). beta-acetoxy cycloart-25-en-24-one (1a) and glutinol (6), lupenone (7), dammaranodienol (9), cycloartenol acetate (10), 24-methylenecycloartanol acetate (11) and beta sitosterol (12), isolated previously, were evaluated for their antiviral activities against Herpes simplex virus (HSV) and African swine fever virus (ASFV). Lupenone exhibited strong viral plaque inhibitory effect against HSV-1 and HSV-2. The in vitro antifungal and antibacterial activities of la, cycloart 23-ene-3beta,25-diol, 3-acetate (3a) and 6-12 were also investigated. PMID- 14526923 TI - Practical experience of the application of ICRP models in internal dose assessment. AB - The introduction of the Ionising Radiations Regulations 1999 in the UK, which came into force on 1 January 2000, led to significant changes in internal dose assessment. Before this date, assessments were based on the methodology from ICRP Publication 26 and, in general, made use of simple models such as those detailed in ICRP Publication 30. However, the introduction of the new Regulations required the use of ICRP Publication 60 methodology, and, at the same time, the latest ICRP biokinetic models were introduced. Many of these newer models were considerably more complex than the ones they replaced. In particular, the use of 'recycling', where activity is constantly recirculated between different organs, meant that the models could not simply be implemented by use of the Skrable formula, as detailed in ICRP Publication 30. This paper outlines two aspects of the application of these latest ICRP models. First, the problems encountered during implementation of these models are detailed, and secondly, it covers the practical experience of using the resulting computer programs for internal dose assessment. PMID- 14526924 TI - Target cells in internal dosimetry. AB - Data related to radium induced bone sarcomas in humans are used as a method of defining target cells on bone surfaces and in the bone marrow. The differential distribution of radiation induced bone sarcoma types, with a high ratio of non bone producing, mainly fibroblastic tumours, challenges the ICRP concept that the bone lining cells are target cells. Multipotential mesenchymal stem cells are located within the range of alpha particles and are the most likely target cells for the fibroblastic type of bone sarcoma. The histogenesis of bone sarcomas after irradiation with alpha emitters shows that their final histopathology is not dependent on a single target cell. Each target cell has a microenvironment, which has to be regarded as a synergistic morpho-functional tissue unit. For this the concept of 'histion', a term used in general pathology, is proposed. Interactions between target cells that have been hit by alpha particles, leading to lethal, mutational or transformation events with all components of a 'histion', will prove critical to understanding the pathogenesis of both deterministic and stochastic late effects. PMID- 14526925 TI - A new model for the human alimentary tract: the work of a Committee 2 task group. AB - The absorbed doses received from ingested radionuclides are currently calculated using the method outlined in ICRP Publication 30. It has been recognised by the ICRP that some of the assumptions used to calculate absorbed doses should be reviewed. A new physiologically based model has been developed, applicable to intakes of radionuclides in food and liquids by children and adults. All parts of the alimentary tract are included, additional sites for absorption and retention are considered and the morphometric and transit parameter values are reviewed in coordination with the ICRP Reference Man revision. Features of the new model are explained and preliminary dose estimates are presented. PMID- 14526926 TI - Doses and risks from the ingestion of Dounreay fuel fragments. AB - The radiological implications of ingestion of nuclear fuel fragments present in the marine environment around Dounreay have been reassessed by using the Monte Carlo code MCNP to obtain improved estimates of the doses to target cells in the walls of the lower large intestine resulting from the passage of a fragment. The approach takes account of the reduction in dose due to attenuation within the intestinal wall and self-absorption of radiation in the fuel fragment itself. In addition, dose is calculated on the basis of a realistic estimate of the anatomical volume of the lumen, rather than being based on the average mass of the contents, as in the current ICRP model. Our best estimates of doses from the ingestion of the largest Dounreay particles are at least a factor of 30 lower than those predicted using the current ICRP model. The new ICRP model will address the issues raised here and provide improved estimates of dose. PMID- 14526927 TI - Uranium gastrointestinal absorption: the f1 factor in humans. AB - The present investigation was undertaken by the Department of Health, Canada, to determine the most appropriate value to use for uranium gastrointestinal absorption (f1) in setting the guideline for drinking water. Fifty participants, free from medical problems, were recruited from two communities: a rural area where drinking water, supplied from drilled wells, contained elevated levels of uranium and an urban area where the water supplied by the municipal water system contained < 1 microg l(-1). Uranium intake through food, drinking water and other beverages was monitored using the duplicate diet approach. Intake and excretion were measured by inductively coupled plasma-mass spectrometry (ICP-MS) in samples collected concurrently from the same individuals over a 3 d period. The range of f1 values was between 0.001 and 0.06, with a median of 0.009. These values were independent of gender, age, duration of exposure, daily total uranium intake and allocation of intake between food and water. Consistent with the recommendation of ICRP Publication 69, 78% were below 0.02. PMID- 14526928 TI - Intake and excretion of 99Tc in a common sea alga, hijiki (Hizikia fusiforme), as ingested in the daily diet--a measurement in an adult male. AB - Ingestion and excretion of environmental 99Tc was measured in an adult male who ingested 'hijiki' food in his diet that contained a sub-bequerel activity per kg, and which was prepared from a marine alga, Hizikia fusiforme, or hijiki in Japanese, traditionally eaten in the Japanese diet. Harvested hijiki was boiled, dried and cooked. This was eaten at each meal five times over 2 d. All faeces and 24-h urine samples were collected during the 3 d before and subsequent 11 d starting from the day when the intake began. The ingestion intake was measured by analysing the dried hijiki material. 99Tc was determined by using liquid-liquid extraction and low-background beta ray spectrometry. The excreta were also analysed for minerals by atomic absorption spectrometry. The apparent absorption rate in the gastrointestinal tract for 99Tc under these conditions was calculated to be approximately 0.5. The present result from a single subject indicates the need for further studies. PMID- 14526929 TI - Bio-kinetics of radon ingested from drinking water. AB - Previous studies have identified the stomach as the most significant organ for the dose from ingested radon. An important factor in dosimetric modelling is the rate of radon loss from the stomach. In the present study, two subjects who ingested radon-rich water were measured using a NaI(Tl) detector fixed over the stomach. The counting rates for 214Pb and 214Bi peak regions were plotted as a function of time after ingestion. These data were interpreted using a compartment model that expressed biokinetics of radon and its progeny. The model was fitted to the experimental data by changing biokinetic parameters such as the rate of radon loss from the stomach. Previous models for dosimetric purposes often assumed that the half-time for radon loss from the stomach is below 20 min. The present results, however, suggest that a part of radon stayed longer in the stomach than expected in the previous models. PMID- 14526930 TI - Practical application of the ICRP Human Respiratory Tract Model. AB - The ICRP Publication 66 Human Respiratory Tract Model (HRTM) has been applied to calculate dose coefficients and bioassay functions using default values of parameters relating to the material and the subjects. The ICRP Task Group on Internal Dosimetry (INDOS) has developed a guidance document on application of the HRTM in situations where using specific information can improve dose assessments. INDOS is now revising the worker exposure documents (ICRP Publications 68 and 78). Application of the HRTM requires a review of the lung-to blood absorption characteristics of inhaled radionuclides. Where appropriate, compound-specific absorption parameter values will be derived, and other compounds will be assigned to default Types using current information. Although no major changes to the HRTM are envisaged, this revision provides an opportunity for some refining and updating in the light of experience and new information. PMID- 14526931 TI - Stochastic model of ultrafine particle deposition and clearance in the human respiratory tract. AB - Deposition and clearance of insoluble ultrafine particles, ranging from 1 to 100 nm, were simulated by stochastic models using Monte Carlo methods. Brownian motion is the dominant mode of deposition in human airways. The additional effects of convective diffusion in bifurcations and axial diffusion (convective mixing) primarily affect particle transport and deposition of particles in the 1 10 nm range. Regarding total deposition, the effects of both convective mechanisms are practically compensated by the concomitant effect of molecular radial diffusion (Brownian motion). During the first hours following inhalation, 1 nm particles are predicted to be cleared much faster than particles in the size range from 10 to 100 nm, with a retained fraction of about 80% after 24 h. For 1 10 nm particles, extracellular transfer to blood is the most likely mode of clearance, while uptake and subsequent accumulation in epithelial cells are assumed to be the preferential mechanisms for 10-100 nm particles. PMID- 14526933 TI - Modifying the ICRP 66 dosimetry model based on results obtained from Mayak plutonium workers. AB - Results obtained in a study of the microscopic distribution of plutonium in the lungs of deceased Pu workers from the Mayak Production Association showed that the long-term retention of Pu was greater than predicted by the current ICRP 66 respiratory tract dosimetry model (HRTM). These data were therefore applied to the HRTM by modifying selected parameters, namely the transfer rate of Pu from the transformed state compartment and the fraction of Pu that transfers to the bound state compartment. Invoking the latter compartment into the modelling allowed a better representation of the long-term Pu retention as well as providing a convenient means of describing the workplace-specific characteristics of the different Pu aerosols found in the Mayak plant. In particular, the present model describes a significantly greater long-term retention of Pu nitrate aerosols in the lung compared with the Type M default. PMID- 14526932 TI - Distribution of plutonium particles in the lungs of Mayak workers. AB - Lung tissues from workers at the Mayak Production Association were examined to determine the distribution of plutonium (Pu) activity in various lung compartments. Stereological sampling methods and autoradiography were used. Pu particles were identified by microscopic examination of autoradiographs and localised in one of six normal anatomic sites and two sites of fibrosis (parenchymal, non-parenchymal). Particle activity was determined by counting the number of tracks emanating from the particles. Over 50% of the Pu activity was localised in sites of fibrosis, which had significantly higher than average activity for the lung. Over 40% of the activity was in lung parenchyma. Activity in the bronchovascular interstitium was significantly lower than average. These results support the hypothesis that Pu activity is not uniformly distributed in the lung, with long-term retained particles concentrated in scars of the lung. The results may significantly affect estimates of dose from inhaled Pu. PMID- 14526935 TI - Assessment of intakes of mixtures of radionuclides. AB - Australia has several uranium mines and a large number of mineral sand mines, with associated processing facilities. Exposures resulting from these mining and processing operations usually involve intakes of mixtures of radionuclides. This work describes the development of a suite of first order, linear compartment models, based on the ICRP Publication 66 respiratory tract model, and an analytical solution to the decay equations, for assessing the consequences of such intakes. The computer programs based on these models directly compute excretion, organ retention and organ and whole-body doses for intakes of either single radionuclides or any mixture of radionuclides belonging to the same radioactive decay chain. The intake can be via inhalation, ingestion or injection, and can be acute, chronic or of limited duration. The starting concentration and degree of secular (dis)equilibrium can be specified for each radionuclide. No assumptions need to be made about the relative magnitudes of the radioactive half-lives of the different nuclides. PMID- 14526934 TI - Comparison of predicted with observed biokinetics of inhaled plutonium nitrate and gadolinium oxide in humans. AB - The absorption kinetics to blood of plutonium and gadolinium after inhalation as nitrate and oxide in humans and animals has been studied. For each material, values describing the time dependence of absorption were derived from the studies in animals and used with the ICRP human respiratory tract model to predict lung retention and cumulative amounts to blood for the volunteers inhaling the same materials. Comparison with the observed behaviour in the volunteers suggests that absorption of plutonium and gadolinium is reasonably species independent, and that data obtained from animal studies can be used to assess their biokinetic behaviour in humans. PMID- 14526936 TI - Mechanistic interpretation of the slow bronchial clearance phase. AB - A slow bronchial clearance phase, describing the removal of insoluble particles from the sol layer, has recently been incorporated into the ICRP human respiratory tract model. A striking feature of that clearance phase is the size dependence of the slowly cleared fraction. In the theoretical approach presented here, it is suggested that the slow bronchial clearance phase includes three main mechanisms: (1) re-transfer of particles onto the gel layer after a certain time delay, (2) intra- or intercellular transfer of particles through the airway epithelium and subsequent transport to the blood or lymph nodes, and (3) uptake of particles by airway macrophages. The experimentally observed inverse relationship between the slow clearance fraction and geometric particle size is interpreted as an increasing capture probability in the sol layer and transepithelial transport with decreasing particle diameter. PMID- 14526939 TI - Biokinetics and assessment of intake of thorium dioxide. AB - The aim of this work was to investigate the biokinetics of thorium dioxide in animals for the purpose of assessing intakes of the compound by workers and the resulting doses. The results imply that measurements of the decay products in the chest or extrapolations from urine analysis data are unlikely to be of value for doses below 20 mSv. Even higher doses should be interpreted with caution as a consequence of uncertainties in particle size distribution and variations in dietary excretion. PMID- 14526937 TI - Comparison of absorption after inhalation and instillation of uranium octoxide. AB - Values for the absorption parameters were compared after inhalation or intratracheal instillation of 1.5 microm mass median aerodynamic diameter (MMAD) 233U3O8 particles into the lungs of HMT strain rats. The two sets of parameter values were similar, as were the calculated dose coefficients and predicted biokinetics for workers. Hence the inhalation and instillation techniques can probably both be used to generate values of the absorption parameters for U3O8. PMID- 14526938 TI - Optimising monitoring regimens for inhaled uranium oxides. AB - This paper provides guidance on the most appropriate monitoring procedures and intervals, the likely uncertainties in the assessment of intake and recommendations on appropriate investigation levels for repeated exposures to uranium trioxide, octoxide and dioxide of natural composition. PMID- 14526940 TI - Implications of human nasal clearance studies for the interpretation of nose blow and bioassay sample measurements. AB - The Smith-Etherington extrathoracic (ET) clearance model has been developed to describe clearance from the human nasal passage as observed for 1.5, 3 and 6 microm monodisperse particles inhaled by participants while sitting or at light exercise. Model parameter values have been determined for 22 sets of nasal clearance data from which typical nasal clearance parameter values have been derived. In this investigation, the faecal excretion of 5 microm activity median aerodynamic diameter (AMAD) Type S 239Pu was calculated for three sets of model parameter values, i.e. typical values and those determined for two experiments that respectively exhibited particularly rapid and slow clearance. The study has shown that a greater fraction of material deposited in ET is cleared to the GI tract, and that this clearance takes place over a longer period than assumed by the Human Respiratory Tract Model for Radiological Protection. The study has also shown that deliberate nose blow sampling might potentially be developed to determine the order of magnitude of intakes. PMID- 14526941 TI - Assessment of the internal effective dose of workers independent of inhaled particle size (II). AB - For 60Co (Type S) and 54Mn (Type M), for which the whole-body content can be measured, it is possible to minimise the errors of the estimated effective doses caused by uncertainty of the activity median aerodynamic diameter (AMAD) by assuming the AMAD to be 5 microm and by measuring the body content on day 5 after inhalation. For the radionuclides to be measured in the lung content, e.g. 239Pu (Type S), it may be necessary to estimate the AMAD, because the lung burdens on any day are not always proportional to the whole-body content which reflects the effective doses. There is no problem in assuming the AMAD to be 5 microm for external counting and for urinalysis of Type F compounds, because of the rapid absorption of such compounds into the blood and the same biokinetics. The breathing rate is assumed to be 1.2 m3 h(-1). PMID- 14526942 TI - Simulation of deposition and clearance of inhaled particles in central human airways. AB - In this study local distributions of deposited inhaled particles such as radon progenies in realistic human airway bifurcation models of bronchial generations one to six are computed for different geometries, inlet flow profiles, flow rates and particle sizes with computational fluid particle dynamics methods. The movement of the mucus layer in the large central human airways is also simulated by computational fluid dynamic techniques. There is experimental evidence that bronchogenic carcinomas mainly originate at the central zone of the large airway bifurcations, where primary hot-spots of deposition have been found. However, current lung deposition models do not take into consideration the inhomogeneity of deposition within the airways. The inhomogeneous movement of the mucus layer may strongly influence the effect of primary deposition. On the basis of our results, both the deposition and the clearance patterns are highly non-uniform, especially in the vicinity of the carinal ridge of the bifurcations. PMID- 14526943 TI - Two case studies of highly insoluble plutonium inhalation with implications for bioassay. AB - Two well characterised Pu inhalation cases show some remarkable similarities between substantially different types of Pu oxide. The circumstances of exposure, therapy, bioassay data, chemical solubility studies and dosimetry associated with these cases suggest that highly insoluble Pu may be more common than previously thought, and can pose significant challenges to bioassay programmes. PMID- 14526945 TI - Deposition and clearance for radon progeny in the human respiratory tract. AB - In vivo counting of 214Pb was conducted to estimate the deposition and retention of radon progeny in the human respiratory tract. Two volunteer subjects were exposed to high radon concentrations. After the exposures, activity deposited in the extrathoracic (ET) region for each subject was measured using a NaI(Tl) detector. According to the International Commission on Radiological Protection (ICRP) model, a reference value for particle transport rate from ET2 to the GI tract is 100 d(-1) (half-time, 10 min). The effective half-time of 214Pb deposited in the ET region was calculated for pure nose and mouth breathers, using the ICRP reference transport rate. While the measured half-times for nose breathers were generally consistent with the calculated values, those for mouth breathers were significantly larger than the calculated values. The results indicated that the particle transport rate from ET2 to the GI tract was much smaller than the reference value in the ICRP model. PMID- 14526944 TI - Sizing alpha emitting particles of aged plutonium on personal air sampler filters using CR-39 autoradiography. AB - Methods have been developed to assess the size distribution of alpha emitting particles of reactor fuel of known composition captured on air sampler filters. The sizes of uranium oxide and plutonium oxide particles were determined using a system based on CR-39 solid-state nuclear track detectors. The CR-39 plastic was exposed to the deposited particles across a 400 microm airgap. The exposed CR-39 was chemically etched to reveal clusters of tracks radially dispersed from central points. The number and location of the tracks were determined using an optical microscope with an XY motorised table and image analysis software. The sample mounting arrangement allowed individual particles to be simultaneously viewed with their respective track cluster. The predicted diameters correlated with the actual particle diameters, as measured using the optical microscope. The efficacy of the technique was demonstrated with particles of natural uranium oxide (natUO2) of known size, ranging from 4 to 150 microm in diameter. Two personal air sampler (PAS) filters contaminated with actinide particles were placed against CR-39 and estimated to have size distributions of 0.8 and 1.0 microm activity median aerodynamic diameter (AMAD). PMID- 14526946 TI - Does mean lung dose calculated after inhalation of alpha emitters actually reflect the risk of induction of malignant lung tumours? AB - A comparison of incidence of lung tumours in rats after inhalation exposure to aerosols containing alpha emitters which have different physico-chemical properties has been performed. Aerosols of radon and progeny, uranium ore dust, NpO2, PuO2 or Cm2O3 were considered for intercomparison with similar or different particle sizes. Dose-effect relationships for the frequency of malignant lung tumours appear linear up to a few Gy and then become infralinear at higher doses delivered to the lungs. The initial slope of the curves reflects the risk of induction of a lung tumour. The highest slopes of incidence were observed for radon and uranium ore dust (about 70 and 20% Gy(-1) respectively) for which the most homogeneous alpha dose distribution to the lungs is expected. In a general trend, increasing the alpha-activity of deposited particles (higher specific activity of constituent radioisotopes or larger particle size) decreases the risk. The comparison of the reported data shows that the risk per Gy at 'low doses' could vary over more than one order of magnitude depending on the physico chemical properties of the aerosols. PMID- 14526947 TI - In vivo measurement of Pu dissolution parameters of MOX aerosols and related uncertainties in the values of the dose per unit intake. AB - The aim of this study was to compare dissolution parameter values for Pu from industrial MOX with different Pu contents. For this purpose, preliminary results obtained after inhalation exposure of rats to MOX containing 2.5% Pu are reported and compared to those obtained previously with MOX containing 5% Pu. Dissolution parameter values appear to increase when the amount of Pu decreases. Rapid fractions, f(r), of 4 x 10(-3) (s.d. = 2 x 10(-3)) and 1 x 10(-3) (s.d. = 6 x 10( 4)) and slow dissolution rates, s(s) of 2 x 10(-4) d(-1) (standard deviation, sigma = 5 x 10(-5)) and 5 x 10(-5) d(-1) (sigma = 1 x 10(-5)) were derived for MOX containing 2.5 and 5% of Pu, respectively. Simulations were performed to assess uncertainties on dose due to experimental errors. The relative standard deviations of the dose per unit intake (DPUI) due to f(r) (4-8%), are far less than those due to s(s) (about 20%), which is the main parameter altering the dose. Although quite different dissolution parameter values were derived, similar DPUIs were obtained for MOX aerosols containing 2.5 and 5% Pu which appear close to that for default Type S values. PMID- 14526948 TI - Assessment of exposure to depleted uranium. AB - In most circumstances, measurement of uranium excreted in urine at known times after exposure is potentially the most sensitive method for determining the amount of depleted uranium (DU) incorporated. The problems associated with this approach are that natural uranium is always present in urine because of the ingestion of natural uranium in food and drink, and that the uncertainties in the intakes as assessed from excretion measurements can be quite large, because many assumptions concerning the exposure characteristics (time pattern of exposure, route of intake, chemical form, solubility, biokinetics within the body) must be made. Applying currently available methods and instruments for the measurement of uranium in urine samples, DU incorporations of levels relevant with respect to potential health hazards can be detected reliably, even a long time after exposure. PMID- 14526949 TI - Characterisation and dissolution of depleted uranium aerosols produced during impacts of kinetic energy penetrators against a tank. AB - Aerosols produced during impacts of depleted uranium (DU) penetrators against the glacis (sloping armour) and the turret of a tank were sampled. The concentration and size distribution were determined. Activity median aerodynamic diameters were 1 microm (geometric standard deviation, sigma(g) = 3.7) and 2 microm (sigma(g) = 2.5), respectively, for glacis and turret. The mean air concentration was 120 Bq m(-3), i.e. 8.5 mg m(-3) of DU. Filters analysed by scanning electron microscopy (SEM) and X ray diffraction showed two types of particles (fine particles and large molten particles) composed mainly of a mixture of uranium and aluminium. The uranium oxides were mostly U3O8, UO2.25 and probably UO3.01 and a mixed compound of U and Al. The kinetics of dissolution in three media (HCO3-, HCl and Gamble's solution) were determined using in-vitro tests. The slow dissolution rates were respectively slow, and intermediate between slow and moderate, and the rapid dissolution fractions were mostly intermediate between moderate and fast. According to the in-vitro results for Gamble's solution, and based on a hypothetical single acute inhalation of 90 Bq, effective doses integrated up to 1 y after incorporation were 0.54 and 0.56 mSv, respectively, for aerosols from glacis and turret. In comparison, the ICRP limits are 20 mSv y(-1) for workers and 1 mSv y(-1) for members of the public. A kidney concentration of approximately 0.1 microg U g(-1) was predicted and should not, in this case, lead to kidney damage. PMID- 14526950 TI - Measuring aerosols generated inside armoured vehicles perforated by depleted uranium ammunition. AB - In response to questions raised after the Gulf War about the health significance of exposure to depleted uranium (DU), the US Department of Defense initiated a study designed to provide an improved scientific basis for assessment of possible health effects on soldiers in vehicles struck by these munitions. As part of this study, a series of DU penetrators were fired at an Abrams tank and a Bradley fighting vehicle, and the aerosols generated by vehicle perforation were collected and characterised. A robust sampling system was designed to collect aerosols in this difficult environment and monitor continuously the sampler flow rates. The aerosol samplers selected for these tests included filter cassettes, cascade impactors, a five-stage cyclone and a moving filter. Sampler redundancy was an integral part of the sampling system to offset losses from fragment damage. Wipe surveys and deposition trays collected removable deposited particulate matter. Interior aerosols were analysed for uranium concentration and particle size distribution as a function of time. They were also analysed for uranium oxide phases, particle morphology and dissolution in vitro. These data, currently under independent peer review, will provide input for future prospective and retrospective dose and health risk assessments of inhaled or ingested DU aerosols. This paper briefly discusses the target vehicles, firing trajectories, aerosol samplers and instrumentation control systems, and the types of analyses conducted on the samples. PMID- 14526951 TI - Excretion of depleted uranium by Gulf War veterans. AB - During the Persian Gulf War, in 1991, approximately 100 US military personnel had potential intakes of depleted uranium (DU), including shrapnel wounds. In 1993, the US government initiated a follow-up study of 33 Gulf War veterans who had been exposed to DU, many of whom contained embedded fragments of DU shrapnel in their bodies. The veterans underwent medical evaluation, whole-body counting, and urinalysis for uranium by kinetic phosphorescence analysis (KPA). Data are available from seven individuals who exceeded the detection limit for whole-body counting and also had elevated urinary uranium. Urinary excretion rates, in microg U g(-1) creatinine, were determined in 1997 and 1999. The body contents, in mg DU, were determined in 1997; it is assumed there were no significant decreases in total body content in the interim. For the 1997 data, the mean fractional excretion was (2.4 +/- 2.8) x 10(-5) g(-1) creatinine, and for the 1999 data, the mean was (1.1 +/- 0.6) x 10(-5) g(-1) creatinine. However, these means are not significantly different, nor is there any correlation of excretion rate with body content. Thus, human data available to date do not provide any basis for determining the effects of particle surface area, composition and solubility, and biological processes such as encapsulation, on the excretion rate. PMID- 14526952 TI - Anomalies between radiological and chemical limits for uranium after inhalation by workers and the public. AB - Exposure limits for workers and the public are based on both chemical toxicity and radiation dose. As a consequence of the different procedures used in their calculation they are incompatible, and adherence to one limit may result in a serious breach of the other. This paper explores the background to these limits, the problems posed by their application and proposes how best to achieve compliance with both limits. PMID- 14526953 TI - The United States Transuranium and Uranium Registries as sources for actinide dosimetry and bioeffects. AB - The United States Transuranium and Uranium Registries (USTUR) has analysed tissues collected at autopsies of over 300 former radiation workers from actinide processing sites throughout the US, in addition to collecting the medical and radiation exposure histories of those workers. These data are included in a large USTUR database and they are available to research scientists throughout the world, either as public records or through collaborative projects with the USTUR. The USTUR also operates the National Human Radiobiological Tissue Repository (NHRTR), in which portions of tissue samples collected at autopsy are kept. These samples, frozen at -70 degrees C, may be used for molecular studies of the effects of radiation. Medical and radiation exposure histories of the tissue donors are available, as are the results of radiochemical analyses of adjacent portions of the samples. These materials are available to researchers who have collaborative agreements with the USTUR, which can be established by accessing USTUR staff members through the website, http://www.ustur.wsu.edu. PMID- 14526954 TI - Structure of a physiologically based biokinetic model for use in 14C and organically bound tritium dosimetry. AB - Physiologically based biokinetic (PBBK) dosimetry models for beta emitters such as 3H and 14C must include rapid turnover compartments which, while they may be minor in terms of dose commitment, can dominate bioassay measurements at early times after intake. In this paper, a consistent PBBK model structure will be described for use in dose assessments for organic 14C and organically bound tritium (OBT), and also for 14CO2, based on the literature of human carbon metabolism, and on direct measurements of human excretion. CO3/HCO3- is a central compartment in carbon metabolism. The 14CO2 biokinetic model described in ICRP Publication 80 for the calculation of dose coefficients was found to omit early components of excretion necessary for the accurate interpretation of bioassay results. Recommendations on the requirements on dosimetry models for intakes of 14C and OBT are made. PMID- 14526956 TI - Validation and verification of the ICRP biokinetic model of 32P: the criticality accident at Tokai-Mura, Japan. AB - Regrettably, a criticality accident occurred at a uranium conversion facility in Tokai-mura, Ibaraki, Japan, on 30 September 1999. Radioactivities of 32P in urine, blood and bone samples of the victims, who were severely exposed to neutrons, were measured. 32P was induced in their whole bodies at the moment of the first nuclear release by the reaction 31P (n, gamma) 32P and 32S (n, p) 32P. A realistic biokinetic model was assumed, as the exchange of 32P between the extracellular fluid compartment and the soft tissue compartment occurs only through the intracellular compartment, and the model was used for preliminary calculations. Some acute excretion of 32P, caused by decomposition or elution of tissues which occurred at the time of the accident, may have happened in the victims' bodies in the first few days. The working hypotheses in the present work should initiate renewed discussion of 32P biokinetics. PMID- 14526955 TI - A generic biokinetic model for predicting the behaviour of the lanthanide elements in the human body. AB - Information on the biokinetics of the 15 elements of the lanthanide series, 57La to 71Lu, is too sparse to permit individual development of meaningful biokinetic models to describe the behaviour of each of the elements in humans. The lanthanides show a regular gradation in chemical properties across the series, and animal studies indicate that this is reflected in regular differences in their deposition in tissues such as the liver and skeleton. These regular differences in chemical and biological behaviour have been utilised to construct a generic lanthanide biokinetic model and to define element-specific parameters for each element in the series. This report describes the use of the available biokinetic data for humans and animals to derive the parameters for each of the elements. PMID- 14526958 TI - Scientific basis for the development of biokinetic models for radionuclide contaminated wounds. AB - Radionuclide-contaminated wounds are of radiological concern because the wound provides a portal of entry of the radionuclide to the systemic circulation, and can also be a tissue at risk if sufficient dose is deposited at the wound site. Accordingly, a scientific committee established jointly by the US National Council on Radiation Protection and the International Commission on Radiological Protection has been developing an approach to describing the biokinetics of radionuclides deposited in wounds and calculating dose to the wound site. This paper focuses on the analyses, performed principally using experimental animal data, that have led to the development of a biokinetic model for deposited soluble radionuclides as well as more insoluble forms, such as colloids, particles and fragments. The available data for injected soluble materials have provided a basis for categorising 48 different elements (from Be to Cm and representing all of the chemical groups, except halogens and noble gases) into four distinct retention groups. In general, the data are adequate for developing a mechanistically based biokinetic model, whose application is exemplified for soluble radionuclides. PMID- 14526957 TI - Stable tracer investigations in humans for assessing the biokinetics of ruthenium and zirconium radionuclides. AB - The interest in the biokinetics of ruthenium and zirconium in humans is justified by the potential radiological risk represented by their radionuclides. Only a few data related to the biokinetics of ruthenium and zirconium in humans are available and, accordingly, the biokinetic models currently recommended by the ICRP for these elements are mainly based on data from animal experiments. The use of stable isotopes as tracers, coupled with a proper analytical technique (nuclear activation analysis with protons) for their determination in biological samples, represents an ethically acceptable methodology for biokinetic investigations, being free from any radiation risk for the volunteer subjects. In this work, the results obtained in eight biokinetic investigations for ruthenium, conducted on a total of three healthy volunteers, and six for zirconium, performed on a total of three subjects, are presented and compared to the predictions of the ICRP models. PMID- 14526959 TI - Application of new imaging and calculation techniques to activity and dose assessment in the case of a 106Ru contaminated wound. AB - The aim of this paper is to describe the dosimetric evaluation of a point contamination that occurred in a laboratory during the examination of an irradiated sample. The incident led to point contamination of the operator's finger due to the presence of mainly 106Ru, with its progeny, 106Rh. The paper reports on the activity and dose assessment, performed using several methods. The measured activity was obtained using a conventional device based on a germanium detector and confirmed using software developed at IRSN, based on reconstruction of voxel phantom associated with the Monte Carlo N-Particle code (MCNP) for in vivo measurement. Two dose assessment calculations were performed using both analytical and Monte Carlo methods, applying the same approach as for activity assessment based on the personal computational phantom of the finger. The results are compared, followed by a discussion on the suitability of the tools described in this study. PMID- 14526960 TI - A biokinetic model for predicting the retention of 3H in the human body after intakes of tritiated water. AB - Scarce published data on the long-term excretion of tritiated water from the human body have been re-evaluated in order to develop a biokinetic model describing the retention in the human body of 3H from tritiated water (HTO) that could be used for both prospective and retrospective radiation protection. A three-component exponential function is proposed to describe the elimination of 3H from HTO with biological half-times of 10 d (99.00%), 40 d (0.98%) and 350 d (0.02%) respectively. The model predicts a committed effective dose of 1.7 x 10( 11) Sv Bq(-1), comparable with that of the current ICRP Publication 56 and 72 models, and estimates the retention of 3H to within a factor of about 2 of the measured values up to 40 d after intake and about 5 at times longer than 100 d. The derivation of the model and the uncertainties associated with the various parameters are discussed. PMID- 14526961 TI - The effect of state of health on organ distribution and excretion of systemic plutonium in the Mayak workers. AB - The extrapulmonary distribution of plutonium in 20 organs (excluding the respiratory tract) was studied in workers who chronically inhaled plutonium at the radiochemical plants of the Mayak Production Association (Ozyorsk, Russia). The data were obtained by radiochemical analysis of soft tissue and bones samples collected at autopsy of 591 workers. The systemic plutonium distribution was determined in healthy individuals as well as in those with health impairment, specifically for those with liver diseases. Twenty-five years after the beginning of inhalation, systemic fractions in the liver and skeleton of individuals who were healthy at the time of death approximate the ratio 45%:45% proposed in the International Commission on Radiological Protection (ICRP) Publication 30. Pathological processes in the liver, accompanied by fatty dystrophy of hepatocytes, increased plutonium clearance from the liver. There was a considerable shift of the plutonium from the liver to the skeleton in individuals who died from liver disease. The average fractions of systemic plutonium in the liver and skeleton of those individuals were 14% and 78% respectively, which did not correspond to ICRP models, indicating a significant effect of disease conditions. Plutonium that was not redistributed was excreted. The urinary excretion rate of plutonium also correlated with state of health. The observed excretion as a fraction of systemic content was 1.64 x 10(-5) d(-1) for individuals in good health and 2.34 x 10(-5) d(-1) for individuals with various chronic diseases. The current models do not account for the influence of different pathological processes in the body on plutonium distribution and retention in systemic organs. This could have significant consequences for dosimetry calculations and risk estimations. PMID- 14526963 TI - Sensitivity analysis techniques applied to a revised model of molybdenum biokinetics. AB - A revised model of molybdenum biokinetics in humans was recently developed on the basis of experimental data gathered in specific investigations conducted with stable tracers. The model can be used for radiation protection purposes, and it is also a suitable working tool for designing new investigations aimed at further improvements to the model. For the latter goal, a sensitivity analysis was performed in order to determine the most significant model parameters in relation to output measurements performed in studies of molybdenum metabolism. A typical sensitivity analysis approach was adopted, considering the effects in variation of model parameters on the time courses of model outputs such as urinary excretion and blood clearance. A recent new sensitivity technique was considered too, based on the calculation of the so-called generalised sensitivity functions. This combines the sensitivities of the model output with respect to model parameters (as in the typical sensitivity analysis method), with the sensitivities of parameter estimates with respect to changes in model outputs. The results obtained in this analysis suggests that data collected in the first 7 h are critical for the definition of the process of blood clearance and related parameters, whereas reliable information at later times is required for a proper characterisation of urinary excretion. PMID- 14526962 TI - Validation of biokinetic models of Cs and I using their daily dietary intakes and organ contents in an adult Indian population. AB - The daily dietary intakes and total contents of Cs and I in skeletal muscle and thyroid, respectively, for an average Indian adult were estimated by determining their concentrations in the total cooked diet and tissue samples using instrumental neutron activation analysis. These data were then used to predict the biological half-lives of Cs and I and their radioactive counterparts for the Reference Indian Man. The predicted biological half-lives of Cs and I for Reference Indian Man were found to be comparable with the reported values for the ICRP Reference Man. PMID- 14526964 TI - Study of uranium transfer across the blood-brain barrier. AB - Uranium is a heavy metal which, following accidental exposure, may potentially be deposited in human tissues and target organs, the kidneys and bones. A few published studies have described the distribution of this element after chronic exposure and one of them has demonstrated an accumulation in the brain. In the present study, using inductively coupled plasma mass spectrometry (ICP-MS) for the quantification of uranium, uranium transfer across the blood-brain barrier (BBB) has been assessed using the in situ brain perfusion technique in the rat. For this purpose, a physiological buffered bicarbonate saline at pH 7.4 containing natural uranium at a given concentration was perfused. After checking the integrity of the BBB during the perfusion, the background measurement of uranium in control rats without uranium in the perfusate was determined. The quantity of uranium in the exposed rat hemisphere, which appeared to be significantly higher than that in the control rats, was measured. Finally, the possible transfer of the perfused uranium not only in the vascular space but also in the brain parenchyma is discussed. PMID- 14526965 TI - Uncertainties on the committed equivalent dose to the thyroid as a function of age for different iodine isotopes. AB - This study compares uncertainties of equivalent doses after internal contamination by 125I, 129I or 131I. Uncertainties were calculated using reported distributions of physiological parameters and Monte Carlo simulation. In adults, uncertainties increase from 131I to 125I and 129I with 1% of the population receiving 3.9, 4.0 and 7.2 times the median dose for the respective isotopes. In newborns, these values were 7.5, 12.3 and 19.0 for 131I, 125I and 129I respectively. The ratio of the beta dose delivered to the epithelium versus a homogeneous distributed dose was estimated for different iodine concentrations in colloid, epithelium and interstitium. In adults, for 131I, about 40% of the beta dose was delivered to the epithelial cells, whereas this fraction varied depending on the concentration for 125I and 129I, i.e. 20-30% at a relative epithelial concentration of 20% and 7-14% at a concentration of 3%. Small variations were observed depending on age. PMID- 14526966 TI - Infant doses from the transfer of radionuclides in mothers' milk. AB - Assessments of potential internal exposures of the child following radionuclide intakes by the mother require consideration of transfers during lactation as well as during pregnancy. Current ICRP work on internal dosimetry includes the estimation of radiation doses to newborn infants from radionuclides ingested in mothers' milk. Infant doses will be calculated for maternal intakes by ingestion or inhalation of the radionuclides, radioisotopes of 31 elements, for which fetal dose coefficients have been published. In this paper, modelling approaches are examined, concentrating on models developed for iodine, caesium, polonium, alkaline earth elements and the actinides. Comparisons of model predictions show maximum overall transfer to milk following maternal ingestion during lactation of about 30% of ingested activity for 131I, 20% for 45Ca and 137Cs, 10% for 90Sr, 1% for 210Po and low values of less than 0.01% for 239Pu and 241Am. The corresponding infant doses from milk consumption are estimated in preliminary calculations to be about two to three times the adult dose for 45Ca and 131I, 70 80% of the adult dose for 90Sr, about 40% for 137Cs, 20% for 210Po, and <0.1% for 239Pu and 241Am. Infant doses from radionuclides in breast milk are compared with doses to the offspring resulting from in utero exposures during pregnancy. PMID- 14526967 TI - Dose coefficients for the embryo and fetus following intakes of radionuclides by the mother. AB - The International Commission on Radiological Protection has recently issued Publication 88, giving dose coefficients for the embryo, fetus and newborn child from intakes of selected radionuclides of 31 elements by the mother, either before or during pregnancy. The biokinetic models used for calculating these doses were based upon the available human data and the results of animal experiments. This paper summarises the approach used for the development of biokinetic and dosimetric models. It also compares the estimates of dose received by the offspring with those received by the reference adult. The main findings are that, in general, doses to the offspring are similar to or lower than those to the reference adult. For a few radionuclides, however, the dose to the offspring can exceed that to the adult. The reasons for these variations in comparative doses are examined. PMID- 14526968 TI - Dosimetry of radioiodine for embryo and fetus. AB - This paper discusses the biokinetic and dosimetric models adopted in ICRP Publication 88 for the evaluation of fetal doses resulting from maternal intakes of radioiodine. The biokinetic model is used to simulate the behaviour of iodine in both the mother and the fetus. Such simulations provide the basis for the estimation of the dose to the embryo and determine the distribution of maternal iodine at the beginning of the fetal period. The model considers iodine to accumulate in the fetal thyroid from the 11th week. The dose to the fetus delivered following birth is evaluated with the biokinetic and dosimetric models described in ICRP Publication 67. Although a substantial fraction of the emitted energy of electrons and photons is less than 10 keV, conventionally assumed to be non-penetrating radiation, these emissions can escape the small fetal thyroid. Absorbed fractions for both self-dose and crossfire were evaluated for the requirements of radioiodine dosimetry in ICRP Publication 88. PMID- 14526969 TI - Is radiation protection for the unborn child guaranteed by radiation protection for female workers? AB - ICRP Publication 88 recommends doses to the embryo and fetus from intakes of radionuclides by the mother for various intake scenarios. Mainly by answering the question 'Is radiation protection for the unborn child guaranteed by radiation protection for female workers?' it has been assessed if the intake scenarios given in ICRP Publication 88 are adequate for radiation protection purposes. This is generally the case, but the consideration of an additional chronic intake scenario for early pregnancy would be helpful. It is demonstrated that following chronic intake by inhalation, for most radionuclides radiation protection for (female) workers is also adequate for protection of the unborn child, considered as a member of the public. However, there are a number of radionuclides for which possible intakes in routine operations should be more restricted (up to 1% of the annual limits on intake for workers in the case of nickel isotopes) to ensure radiation protection for the unborn child. PMID- 14526970 TI - Transfer of alkaline earth elements in mothers' milk and doses from 45Ca, 90Sr and 226Ra. AB - An international programme of work is currently under way to develop methods for calculating doses to infants from ingestion of radionuclides present in mothers' milk. This paper considers the special case of the alkaline earth elements. Models have been developed for 45Ca, 90Sr and 226Ra and the sensitivity of results to various changes in parameter values is discussed. A complication when calculating doses from intakes of radium is that the International Commission on Radiological Protection has previously recommended that doses from decay products of radium should be calculated using element-specific biokinetic models (so called independent biokinetics). An extension of this method to the models for breastfeeding is proposed. Preliminary estimates of the doses received by the infant for a number of maternal intake scenarios show that doses to the infant can exceed the corresponding adult dose, such as for 45Ca (ratio = 3.1) while, in other cases such as 90Sr, the infant dose can be a significant fraction of the adult dose. PMID- 14526971 TI - Some aspects of the fetal doses given in ICRP Publication 88. AB - The International Commission on Radiological Protection (ICRP) has recently published dose coefficients (dose per unit intake, Sv Bq(-1)) for the offspring of women exposed to radionuclides during or before pregnancy. These dose estimates include in utero doses to the embryo and fetus and doses delivered postnatally to the newborn child from radionuclides retained at birth. This paper considers the effect on doses of the time of radionuclide intake and examines the proportion of dose delivered in utero and postnatally for different radionuclides. Methods used to calculate doses to the fetal skeleton are compared. For many radionuclides, doses are greatest for intakes early in pregnancy but important exceptions, for which doses are greatest for intakes later in pregnancy, are iodine isotopes and isotopes of the alkaline earth elements, including strontium. While radionuclides such as 131I deliver dose largely in utero, even for intakes late in pregnancy, others such as 239Pu deliver dose largely postnatally, even for intakes early during pregnancy. For alpha emitters deposited in the skeleton, the assumption made is of uniform distribution of the radionuclide and of target cells for leukaemia and bone cancer in utero; that is, the developing bone structure is not considered. However, for beta emitters, the bone structure was considered. Both approaches can be regarded as reasonably conservative, given uncertainties in particular in the location of the target cells and the rapid growth and remodelling of the skeleton at this stage of development. PMID- 14526972 TI - Biokinetics of radiotellurium in rats. AB - Radiotellurium is present in the environment primarily due to its release during nuclear reactor accidents. Little is known of tellurium metabolism in juveniles, although the element is relatively abundant and has a number of industrial uses. A biokinetic study of radiotellurium in rats was performed using gamma-ray counting. Wistar strain rats were used to determine the uptake of H2(123m)TeO3 by the whole-body retention of juvenile rats and the conceptus in relation to its gestational stages, by measurements in the placenta, fetal membranes, fetal fluid and fetus. The whole-body retention of 123mTe in juvenile rats was higher than that of adult rats. The relative concentration in the placenta and fetal membranes was higher than in the fetus. No activity was observed in the fetal fluid. These results indicate that the placenta and fetal membranes play significant roles as barriers to the transfer of 123mTe into the fetus. The ratio, relative concentration in fetus/relative concentration in mother (C(F)/C(M)), was calculated. The C(F)/C(M) ratio was dependent on the stage of gestation and ranged from 0.2 to 0.5. A little 123mTe was transferred to the suckling rats through the mother's milk when the isotope was administered intravenously to the mother. PMID- 14526973 TI - Transfer of 14C to prenatal and neonatal rats from their mothers exposed to 14C compounds by ingestion. AB - The transfer of 14C through placenta or milk was investigated and the radiation dose to fetal and newborn rats was estimated. Female rats at different gestational stages or after delivery were exposed to 14C in the form of sodium bicarbonate, thymidine and lysine by a single ingestion. Radioactivity in maternal tissues and conceptuses (placenta, fetal membrane and fetus) and in the newborn was determined at various times after ingestion. After exposure to these 14C compounds, there was no significant difference between the 14C concentration in the fetus and that in the maternal tissues, suggesting that the placenta has no effect in preventing or accelerating the placental transfer of 14C. The concentration and content of 14C in the fetus and newborn were, however, dependent on the chemical form of 14C and on the prenatal or neonatal stage at the time of ingestion. The result of the dose estimation showed that 14C-lysine gave significantly higher prenatal and neonatal doses than 14C-sodium bicarbonate or 14C-thymidine. PMID- 14526974 TI - The ICRP working party on bioassay interpretation. AB - In recent years there have been many developments in modelling the behaviour of radionuclides in the human body. The current generation of models is designed to be more 'realistic' than the previous generation of simple compartment models. The International Commission on Radiological Protection (ICRP) uses these models to produce dose coefficients and recognises that there is a need to give more guidance on how these models can be used to interpret bioassay data. A working party has been set up to address the issue. This paper describes some of the problems, some approaches to solving the problems and the progress of the ICRP working party. PMID- 14526975 TI - Control of the risk of exposure to alpha emitting radionuclides in French nuclear power plants: example of Cattenom. AB - Control of the risk of internal exposure of EDF PWR plant maintenance workers by alpha-emitting radioactive elements is based on identification and quantification of the contamination of the systems. In 2001, an experiment carried out at Cattenom Power Plant during a unit outage in the presence of a leaking fuel cladding, based on measurement of alpha-emitting radioactive elements, made it possible to determine a realistic particle resuspension coefficient. A resuspension coefficient of 10(-6) m(-1) was adopted for operational radiological protection. An appropriate monitoring system for workers was set in place in collaboration with the occupational medicine and radiological protection department. It was based on prior estimation of the level of alpha contamination, and confirmed by swipe measurements, atmospheric surveillance by monitors, and collective analysis by nose blow samples from workers selected on the basis of their workstations, as well as supplementary individual measurements (monitoring of faeces). This surveillance made it possible to validate an appropriate work area monitoring system. PMID- 14526977 TI - NORM workers: a challenge for internal dosimetry programmes. AB - The mining, milling and processing of uranium and thorium bearing minerals may result in radiation doses to workers. The control of occupational exposures from these natural sources of radiation imposes a challenge to regulators and radiation protection advisers. A survey pilot programme, which included six mines in Brazil and a monazite plant, was established, consisting of the collection and analysis of concentrations of uranium, thorium and polonium in urine, faeces and air samples. Results from workers were compared to background data from their families living in the same area and from residents from the population of Rio de Janeiro. Positive exposure results were identified among the coal miners, the niobium miners and the monazite sand workers. Difficulties in the application of internal dosimetry programmes are discussed in relation to the control of NORM workers. PMID- 14526976 TI - Database of calculated values of retention and excretion for members of the public following acute intake of radionuclides. AB - Intakes of radionuclides can be estimated from measurements of radioactivity in the whole body or in specific organs or in excreta by comparing them with predicted retention or excretion data calculated using standard biokinetic models. For occupational exposure monitoring, data are presented by ICRP for 29 radionuclides in Publication 78 (1997) and by the authors for 42 radionuclides as electronic look-up tables in Microsoft Excel. In the present work, values of retention and excretion were computed for selected radionuclides inhaled or ingested by members of the public. Graphs were constructed from the computed results showing the predicted monitoring data as functions of time following acute intakes of radionuclides. A graphical database was assembled on the Web site http//www.nirs.go.jp/RPD/ to provide a tool for the interpretation of bioassay measurements. PMID- 14526978 TI - Anomalously high excretion of Pu in urine following inhalation of plutonium nitrate? AB - A study of the biokinetics of inhaled plutonium nitrate in two volunteers has been carried out. Low doses (approximately 80 microSv) were achievable because tracers of high isotopic purity were used: 237Pu (measurable by X ray spectrometry) and 244Pu (measurable by accelerator mass spectrometry). Lung retention, amount in blood, uptake to the liver and skeleton, and urinary and faecal excretion were measured. The measured urinary excretion rates are about a factor of three higher than those predicted from urine excretion data measured following intravenous injection of plutonium to the same volunteers. If similar biokinetic behaviour occurs in workers exposed to plutonium nitrate, intakes by inhalation and corresponding committed doses assessed by urine bioassay could be consistently overestimated by a similar factor. PMID- 14526979 TI - Assessment of occupational doses from internal contamination with 241Am. AB - A group of workers with occupational intakes of 241Am, which occurred a long time ago, has been followed for some time. Results of in vivo measurement and bioassay of excreta are compared with the values predicted by the ICRP Publication 78 model. The observed skeletal content is, as a rule, higher than the predicted one. The ratio of excreted activity in urine to that in faeces is in very good agreement with the model prediction. Another group of workers from a waste management department, who were internally contaminated in July 2001, has also been followed. In some cases, there is quite a large difference in calculated intake between excretion by urine and that by faeces. The contaminant was presumably the same as that in the group of workers with old intakes, but its physical and chemical form could be influenced by a fixating lacquer used to prevent the spread of contamination. PMID- 14526981 TI - The application of Bayesian techniques in the interpretation of bioassay data. AB - The inverse problem of internal dosimetry is naturally posed as a problem of Bayesian inference. The Bayesian approach is of practical importance in three areas: (1) avoiding false positives in the detection of rare events, (2) the calculation of uncertainties, and (3) the calculation of multiple intakes, all of which are important for internal dosimetry. In this paper, the Bayesian approach to the interpretation of measurements is first reviewed using a simple conceptual example. Then, a simple 239Pu case using IMBA expert is discussed, and finally a current cutting-edge example is discussed involving real 238Pu data calculated with a Markov Chain Monte Carlo algorithm and with exact calculation of poisson likelihood functions. PMID- 14526980 TI - Internal dose assessment in radiation accidents. AB - Although numerous models have been developed for occupational and medical internal dosimetry, they may not be applicable to an accident situation. Published dose coefficients relate effective dose to intake, but if acute deterministic effects are possible, effective dose is not a useful parameter. Consequently, dose rates to the organs of interest need to be computed from first principles. Standard bioassay methods may be used to assess body contents, but, again, the standard models for bioassay interpretation may not be applicable because of the circumstances of the accident and the prompt initiation of decorporation therapy. Examples of modifications to the standard methodologies include adjustment of biological half-times under therapy, such as in the Goiania accident, and the same effect, complicated by continued input from contaminated wounds, in the Hanford 241Am accident. PMID- 14526982 TI - Eliminating bias in routine bioassay when there is an unknown time of intake. AB - Routine bioassay programmes sometimes find evidence of an unsuspected intake. If there were no workplace indicators of exposure or intake, it is necessary to assume a value for the time of intake. Under these circumstances, the International Commission on Radiological Protection (ICRP) continues to recommend using the midpoint of the interval between routine bioassay measurements (ICRP Publication 78, paragraph 106). The assumption of T/2 as the time of intake, where T is the interval between bioassay measurements, represents the expectation value of the time of intake, (t), assuming uniform probability of an intake at any given time. This assumption results in a modest bias, of the expectation value of the intake, (I), that would have been received by a population of workers who had uniform probability over time of intake. This underestimation leads to a negative or positive bias in dose estimates derived in this fashion. The bias is characterised for realistic, routine urinalysis programs for Pu, U and 3H, as well as for in vivo measurements of 125I, 131I and 137Cs. Simple numerical methods are presented for correcting the bias. The bias is greatest for radionuclides whose half-lives are short with respect to the interval between bioassay measurements. Since the primary concern is estimating intake rather than time, the assumed time of intake should be chosen as t(I) rather than T/2. The ICRP should consider revising some of the tables in its Publication 78 to reflect this. PMID- 14526983 TI - Review of methods and computer codes for interpretation of bioassay data. AB - Internal dose determination is an essential component of individual monitoring programmes for workers or members of the public exposed to radionuclides, and methods and computer programs are required for dose assessment. A recent international European Radiation Dosimetry Group (EURADOS) intercomparison has shown unacceptably large ranges in the results assessment. An ICRP working party has been initiated to consider what guidance ICRP can give on the use of models and interpret bioassay data in terms of intake/dose. In this field, six codes for bioassay data interpretation, which implement the current ICRP publication 78 biokinetic models, have been reviewed against several criteria with different levels of importance: minor criteria such as the practical use of the code and the graphical capabilities, and major criteria such as the choice of available parameters, peculiarities of data fitting and interpretation, the choice of biokinetic models and the use of uncertainties. All these criteria were assessed using one artificial set of data and two examples extracted from the previous international EURADOS intercomparison. PMID- 14526984 TI - OMINEX: survey of internal dose monitoring programmes for radiation workers. AB - Monitoring of the workforce in the nuclear industries is carried out primarily in order to demonstrate compliance with European Union (EU) legislation and the Basic Safety Standards for the protection of the health of workers against the dangers arising from ionizing radiation recommended by the International Atomic Energy Agency (IAEA). There is, however, no common strategy for internal dose monitoring programmes currently in use in the EU countries. Surveys have been carried out in which organisations were asked to provide information on the design of their internal dose monitoring programmes and on the costs of these programmes. Information was requested from both EU countries and Associated States. Databases for storage and reporting of all gained information were constructed, and results from the surveys have been compiled. This work was carried out within the EC 5th Framework Programme project, OMINEX, which aims to provide advice and guidance on designing and implementing internal dose monitoring programmes in the workplace in such a way that best use is made of available resources, while minimising costs. PMID- 14526985 TI - Incorporation measurements in Switzerland: the approved internal dosimetry service at PSI. AB - The Swiss radiation protection regulation requires approval of external and internal dosimetry laboratories. The Swiss Personal Dosimetry Ordinance specifies the terms of approval, the survey methods and the standard interpretation to assess the committed effective dose. Specific information is given for 25 radionuclides concerning metabolism, methods of measurement, survey interval and interpretation of the results. On successful completion of a technical audit by an external expert, the dosimetry service of PSI was approved for internal dosimetry by the Swiss authorities by 1 January 2001. The scope of approval includes five different measuring methods for 30 radionuclides. PMID- 14526986 TI - Guidance on individual monitoring programmes for radioisotopic techniques in molecular and cellular biology. AB - The radioisotope techniques used in molecular and cellular biology involve external and internal irradiation risk. The personal dosemeter may be a reasonable indicator for external irradiation. However, it is necessary to control the possible internal contamination associated with the development of these techniques. The aim of this project is to analyse the most usual techniques and to establish programmes of internal monitoring for specific radionuclides (32P, 35S, 14C, 3H, 125I and 131I). To elaborate these programmes it was necessary to analyse the radioisotope techniques. Two models have been applied (NRPB and IAEA) to the more significant techniques, according to the physical and chemical nature of the radionuclides, their potential importance in occupational exposure and the possible injury to the genetic material of the cell. The results allowed the identification of the techniques with possible risk of internal contamination. It was necessary to identify groups of workers that require individual monitoring. The risk groups have been established among the professionals exposed, according to different parameters: the general characteristics of receptor, the radionuclides used (the same user can work with one, two or three radionuclides at the same time) and the results of the models applied. Also a control group was established. The study of possible intakes in these groups has been made by urinalysis and whole-body counter. The theoretical results are coherent with the experimental results. They have allowed guidance to individual monitoring to be proposed. Basically, the document shows: (1) the analysis of the radiosotopic techniques, taking into account the special containment equipment; (2) the establishment of the need of individual monitoring; and (3) the required frequency of measurements in a routine programme. PMID- 14526987 TI - A computational study of the urinary excretion rates for 339Pu using new ICRP internal dosimetry models. AB - A computational study of the urinary excretion rates for 239Pu has been carried out using a methodology which involved the solution of a complete compartmental model describing the biokinetic behaviour of inhaled plutonium aerosols in the human body. The methodology, after proper validation, was applied to investigate the dependence of urinary excretion rates for 239Pu on the transfer rates given in the complete compartmental model. For this purpose, the default values of the transfer/absorption rates were modified by factors of 2 and 4 and urinary excretion rates were computed on 1, 10, 100, 1000 and 10,000 d post-intake. The percentage variations in the urinary excretion rates as a result of the modified transfer rates were computed for exposures to Type M and S aerosols of 239Pu. These results facilitated the identification of parameters significantly affecting the short-term and long-term urinary excretion rates. In addition, time variations of the predicted ratios of 239Pu activity in daily urine to that in blood (excretion ratios) were studied for the three biokinetic models of plutonium: the ICRP 67 model, the modified ICRP 67 model with the compartment STI to urinary bladder removed and Luciani and Polig's model. All the computational results are presented and discussed in this paper. PMID- 14526988 TI - Feasibility studies for assessing internal exposure to 233U. AB - The potential internal occupational exposure encountered as a consequence of the 232Th-233U fuel cycle are likely to arise predominantly from the inhalation of 232Th, 233U and (232Th + 233U) compounds of absorption Types M and S. In the past, although direct and indirect methods for assessments of internal exposure to 232Th and its daughters were developed, standardised and employed, no such attempts have been made with regard to 233U and 233U + 232Th. Therefore, feasibility studies for assessing internal exposure to 233U have been conducted using three methods: urine bioassay, in vivo counting and measurement of thoron gas in the exhaled breath of a worker. This paper describes details of these studies and discusses the results obtained. PMID- 14526989 TI - An intake of americium oxide powder: implications for biokinetic models for americium. AB - A worker inhaled 241AmO2 powder. Air sampling showed low activities but a nose blow revealed 92 Bq. Results from faecal sampling and lung and whole-body monitoring indicated an intake of about 200 Bq, but urine sampling, though commencing only 1 d after intake, showed below-threshold activities (< 0.2 mBq). This conflicts with predictions based on the ICRP Publication 67 biokinetic model for americium and the ICRP Publication 66 model for the human respiratory tract, if default lung parameters are used. PMID- 14526990 TI - A survey on uncertainty in bioassay measurements carried out within the OMINEX project. AB - One of the topics covered by the OMINEX (Optimisation of Monitoring for Internal Exposure) project was to look at the bioassay measurements performed in the European laboratories. Questionnaires on bioassay measurements by alpha spectrometry or inductively coupled plasma-mass spectrometry (ICP-MS) were sent to European laboratories. The main objective was to collect information on analytical procedures and on the uncertainties associated with bioassay results. Alpha spectrometry is the technique most often used for the determination of alpha emitters, such as actinides in biological samples. ICP-MS is not used much as yet for routine measurements of actinides in biological samples, but is very sensitive for measurements of long-lived radionuclides. The different parameters influencing the uncertainties on the results and the minimum detectable amount have been investigated. Optimum conditions for achieving low result uncertainties and high sensitivity are given for alpha spectrometry and ICP-MS. PMID- 14526991 TI - Determination of 238U, 234U, 232Th, 228Th, 228Ra, 226Ra and 210Pb concentration in excreta samples of inhabitants of a high natural background area. AB - The high concentration of uranium and thorium in certain Brazilian areas provides an opportunity to evaluate the radiation exposure due to intake of radionuclides by the populations that live and work in areas with a high natural radiation background. Buena, where this study was conducted, is a small village on the coast in the northern part of Rio de Janeiro State, characterised by the presence of a large deposit of monazite sand. In this paper, the concentrations of 238U, 234U, 232Th, 228Th, 228Ra, 226Ra and 210Pb in faecal samples from inhabitants of this area were determined by a sequential analytical method. The results of the average concentrations in faeces of inhabitants of Buena are 9.4 +/- 3.4 mBq g(ash)(-1) for 238U, 9.2 +/- 4.0 mBq g(ash)(-1) for 234U, 7.0 +/- 4.2 mBq g(ash)( 1) for 232Th, 256.1 +/- 134.6 mBq g(ash)(-1) for 228Th, 335.5 +/- 192.8 mBq g(ash)(-1) for 228Ra, 156.6 +/- 74.1 mBq g(ash)(-1) for 226Ra and 66.7 +/- 17.7 mBq g(ash)(-1) for 210Pb. The results were compared with background concentrations from faecal samples from individuals living in Rio de Janeiro City. For most of the radionuclides analysed, the average concentration in faeces from inhabitants of the high natural radiation background was higher than the concentration found in Rio de Janeiro, considered a 'normal' background area. PMID- 14526992 TI - Uncertainty analysis of the urinary excretion of plutonium. AB - A quantitative estimate of the uncertainty of the urinary excretion of plutonium predicted by available biokinetic models is provided. Urinary excretion is primarily considered here because the monitoring of internal contamination of plutonium mainly relies on measurements of activity in urine samples. A previous paper has identified the most significant transfer rates for urinary plutonium excretion following an acute intake. That analysis is used here as a screening method to reduce the number of model parameters to be considered. A log-normal distribution was assumed for the probability distribution of the model parameters. The spread of the values, represented by the geometric standard deviation (GSD), is explicitly calculated, as few indications of the range of variation of systemic transfer rates are available. Different values for the GSD were considered. Assuming a certain GSD for all the systemic rate constants, random values of the rates were generated (by means of a Monte Carlo simulation with a Latin hypercube sampling scheme) and the resulting predictions of urine bioassay measurements were calculated. The comparison of the mean and variance of the predictions with the available data from several studies performed on different subjects provides information about the GSD of model parameters that represents the intersubject variation of transfer parameters. PMID- 14526993 TI - Towards a model for the dynamic transfer of tritium and carbon in mammals. AB - Available data have been analysed to test the hypothesis that both 3H and 14C transfer in mammals can be accounted for by an understanding of metabolism. Data obtained from various 14C and 3H experiments with rats and sheep have been analysed to assess the multi-component retention function of various organs and identify any relationship between half-times and component contribution. Our hypothesis was that component half-times for 14C and 3H are similar after intakes of organic compounds. Similarities in the tritium and carbon dynamics between rat and sheep were observed supporting the hypothesis. For fast and slow components of muscle half-time, allometric relationships have been derived. The results obtained could be used in the development of a human biokinetic model. PMID- 14526994 TI - Minimum detectable dose as a measure of bioassay programme capability. AB - This paper suggests that minimum detectable dose (MDD) be used to describe the capability of bioassay programmes for which intakes are expected to be rare. This allows expression of the capability in units that correspond directly to primary dose limits. The concept uses the well established analytical statistic minimum detectable amount (MDA) as the starting point, and assumes MDA detection at a prescribed time post-intake. The resulting dose can then be used as an indication of the adequacy or capability of the programme for demonstrating compliance with the performance criteria. MDDs can be readily tabulated or plotted to demonstrate the effectiveness of different types of monitoring programmes. The inclusion of cost factors for bioassay measurements can allow optimisation. PMID- 14526995 TI - Uncertainty analysis in the task of individual monitoring data. AB - Assessment of internal doses is an essential component of individual monitoring programmes for workers and consists of two stages: individual monitoring measurements and interpretation of the monitoring data in terms of annual intake and/or annual internal dose. The overall uncertainty in assessed dose is a combination of the uncertainties in these stages. An algorithm and a computer code were developed for estimating the uncertainty in the assessment of internal dose in the task of individual monitoring data interpretation. Two main influencing factors are analysed in this paper: the unknown time of the exposure and variability of bioassay measurements. The aim of this analysis is to show that the algorithm is applicable in designing an individual monitoring programme for workers so as to guarantee that the individual dose calculated from individual monitoring measurements does not exceed a required limit with a certain confidence probability. PMID- 14526996 TI - 'Dose per unit content' functions: a robust tool for the interpretation of bioassay data. AB - The purposes of this study were to investigate the influence of the consequences of the lack of primary bioassay information and to elaborate approaches which could improve the reliability of dose assessments. The aggregated time-dependent functions 'dose per unit organ (excretion) content' z(t) have been proposed in this study as a convenient and reliable tool for bioassay. The analysis of the variation of z with changes of AMAD has demonstrated the existence of areas of the relative invariance of z, which permits the selection of one (reference) function z for the whole area of stability. Within the framework of such an approach an arbitrary set of bioassay data can be approximated by the linear combination F(t) = sum(i) E(i)/z(t - tau(i)), where F(t) = function of time t, which approximates the observed bioassay time trend; tau(i) = time shift of the acute intake i; E(i) = effective dose, associated with the acute intake i (the two last parameters are results of the approximation procedure). PMID- 14526997 TI - Application of Monte Carlo calculations to calibration of anthropomorphic phantoms used for activity assessment of actinides in lungs. AB - This paper reports on a new utility for development of computational phantoms for Monte Carlo calculations and data analysis for in vivo measurements of radionuclides deposited in tissues. The individual parameters of each worker can be acquired for an exact geometric representation of his or her anatomy, which is particularly important for low-energy gamma ray emitting sources such as thorium, uranium, plutonium and other actinides. The software discussed here enables automatic creation of an MCNP input data file based on computed tomography (CT) scanning data. The utility was first tested for low- and medium-energy actinide emitters on Livermore phantoms, the mannequins generally used for lung counting, in order to compare the results of simulation and measurement. From these results, the utility's ability to study uncertainties in in vivo calibration were investigated. Calculations and comparison with the experimental data are presented and discussed in this paper. PMID- 14526998 TI - Use of group monitoring data in lung dose estimation for intakes of uranium. AB - This paper describes an internal dosimetry program developed for Canadian uranium processing facilities. The recently adopted recommendations of ICRP Publication 60 have made it extremely difficult to detect intakes of insoluble forms of natural uranium by in vivo methods (lung counting). A type S intake of UO2 corresponding to a 20 mSv effective dose has a lung burden that is a factor of 2 3 lower than the MDA, 6 months after the intake occurred. A new methodology, approved in principle by the Canadian Nuclear Safety Commission, has been designed to overcome this problem by summing sequential, but separate, lung counts for an individual, or summing lung counts from a group of workers performing similar tasks. This summing technique effectively increases the counting time and, therefore, reduces the MDA to a value below the dose limit. Doses will be assigned to the individual or work group based on the average lung burden. PMID- 14526999 TI - Internal dosimetry of uranium isotopes using Bayesian inference methods. AB - A group of personnel at Los Alamos National Laboratory is routinely monitored for the presence of uranium isotopes by urine bioassay. Samples are analysed by alpha spectroscopy, and the results are examined for evidence of an intake of uranium. Because the measurement uncertainties are often comparable to the quantities of material we wish to detect, statistical considerations are crucial for the proper interpretation of the data. The problem is further complicated by the significant, but highly non-uniform, presence of uranium in local drinking water and, in some cases, food supply. Software originally developed for internal dosimetry of plutonium has been adapted to the problem of uranium dosimetry. The software uses an unfolding algorithm to calculate an approximate Bayesian solution to the problem of characterising any intakes which may have occurred, given the history of urine bioassay results for each individual in the monitored population. The program uses biokinetic models from ICRP Publications 68 and later, and a prior probability distribution derived empirically from the body of uranium bioassay data collected at Los Alamos over the operating history of the laboratory. For each individual, the software creates a posterior probability distribution of intake quantity and solubility type as a function of time. From this distribution, estimates are made of the cumulative committed dose (CEDE) to each individual. Results of the method are compared with those obtained using an earlier classical (non-Bayesian) algorithm for uranium dosimetry. We also discuss the problem of distinguishing occupational intakes from intake of environmental uranium, within a Bayesian framework. PMID- 14527000 TI - Blood tritium level as an estimate of soft tissue dose. AB - To test the blood tritium level as an estimate of free water tritium (FWT) and organically bound tritium (OBT) in human soft tissues, a tritiated amino acid mixture (TAA) and tritiated water (HTO) were injected or orally administered to mice. Relatively high tritium retention in the blood was found after TAA injection. This observation was explained by a low rate of clearance of non volatile tritium components. The dose estimated for the soft tissues after TAA injection was larger than that estimated on the basis of tritium concentration in urine by a factor of 1.8-2.3. In both HTO and TAA injections, the blood tritium level gave a conservative estimate of the dose to soft tissues including active bone marrow. The accumulated dose to soft tissues after oral administration tended to be lower than that for intraperitoneal injection. PMID- 14527001 TI - IMBA Expert: internal dosimetry made simple. AB - In 1997, a collaboration between British Nuclear Fuels plc (BNFL), Westlakes Research Institute and NRPB started, with the aim of producing IMBA (Integrated Modules for Bioassay Analysis), a suite of software modules that implement the new ICRP models for estimation of intakes and doses. This was partly in response to new UK regulations, and partly due to the requirement for a unified approach in estimating intakes and doses from bioassay measurements within the UK. Over the past 5 years, the IMBA modules have been developed further, have gone through extensive quality assurance, and are now used for routine dose assessment by approved dosimetry services throughout the UK. More recently, interest in the IMBA methodology has been shown by the United States Department of Energy (USDOE), and in 2001 an ambitious project to develop a software package (IMBA Expert USDOE Edition) which would meet the requirements of all of the major USDOE sites began. Interest in IMBA Expert is now being expressed in many other countries. The aim of this paper is to outline the origin and evolution of the IMBA modules (the past); to describe the full capabilities of the current IMBA Expert system (the present) and to indicate possible future directions in terms of capabilities and availability (the future). PMID- 14527002 TI - Lessons learned from interlaboratory comparisons of bioassay data interpretation. AB - When a set of bioassay data is given to two different dosimetrists, it is likely that these data will be interpreted differently, that different methods and dosimetric models will be applied and therefore different numerical values will be obtained. Thus, it is important for laboratories dealing with internal dosimetry to undergo performance testing procedures such as interlaboratory comparisons of bioassay data interpretation. Several intercomparison exercises have already been organised at national and international levels. The largest one so far was the 3rd European Intercomparison Exercise on Internal Dose Assessment, which has been organised in the framework of the EULEP/EURADOS Action Group, 'Derivation of parameter values for application to the new model of the human respiratory tract for occupational exposure'. The most important lesson learned from these intercomparison exercises was the need to develop agreed guidelines for internal dose evaluation procedures to promote harmonisation of assessments between organisations and countries. PMID- 14527004 TI - Requirements for authorisation of internal dosimetry services. AB - In order to ensure that a facility is in compliance with the occupational exposure requirements established by regulatory authorities, the measurements and dose assessments specified in the individual monitoring programme need to be reliable. There are two important questions that shall be addressed here: one is how the licensed facilities can demonstrate to their workers and regulatory bodies compliance with the regulatory limits and the reliability of the results of the individual monitoring programmes; the other concerns the mechanisms used to demonstrate to a facility in another country the reliability of the measurement results of an individual monitoring bioassay programme. The accreditation of the bioassay laboratory, according to ISO/IEC 17025, shall be the basic requirement for obtaining the authorisation granted by the national regulatory authority. For the second question, such confidence can be achieved through International Laboratory Accreditation Cooperation (ILAC). PMID- 14527003 TI - Present and future activities of the IAEA on internal dosimetry: lessons learned from international intercomparisons. AB - The International Atomic Energy Agency (IAEA) conducts safety activities to support the assessment of occupational exposure due to intakes of radionuclides; a comprehensive set of safety documents will soon be completed. In recent years, extensive improvements in measurement techniques, phantoms and computational tools have been made. Thus, it is important for laboratories involved in internal dosimetry to undergo performance testing procedures to demonstrate the correctness of the methods applied and also to determine the consistency of their results with those obtained by other laboratories. Several intercomparisons were organised, and they revealed significant differences among laboratories in their approaches, methods and assumptions, and consequently in their results. This paper presents the current and future IAEA activities in support of assessment of occupational exposure due to intakes of radionuclides in the IAEA Member States, as well as the lessons learned from several intercomparison exercises in the last 5 years. PMID- 14527005 TI - Italian intercomparison exercise on internal dose assessment. AB - In 2001, the Radiation Protection Institute of ENEA promoted an Italian intercomparison exercise on internal dose assessment addressed to the qualified experts in radiation protection, following the coming into force in Italian law of the EURATOM 96/29 Directive. Five case studies of occupational exposure related to the Italian situation are used. The considered radioisotopes are: 60Co, 89Sr, 125I, 131I, and 222Rn + NORM (238U-235U-232Th). Data related to WBC, thyroid and urine excretion measurements, as well as radionuclide air concentration in the workplace, are provided to the participants. The results related to medical, industrial and Rn occupational exposure are well represented as means of log-normal distributions with values of the geometric standard deviation less than 2. A wider spread of results is present for the evaluation of occupational exposure to NORM. PMID- 14527006 TI - PROCORAD's international intercomparisons highlight the evolution of techniques used to determine uranium in urine. AB - Inter-laboratory tests are a means of assessing the analytical coherence of medical laboratories. In radiotoxicology, this kind of exercise makes it possible to keep up with laboratory know-how and with the evolution and relative performances of analytical techniques. The intercomparison exercises organised by PROCORAD (Association for the Promotion of Quality Controls in Radiotoxicological Bioassays) provide an opportunity to compare and contrast radiochemistry and metrology for the in vitro analysis of urine and faeces. For uranium compounds, the development of new techniques such as inductively coupled plasma-mass spectrometry (ICP-MS), alpha spectrometry, and laser spectrofluorimetry makes it possible to compare the effectiveness of these protocols with respect to radiation protection monitoring, both in routine and special situations. Detection limits, flexibility, repeatability, reproducibility and isotopic quantification are the criteria considered in this study. The authors present the evolution of performances for the analysis of uranium in urine over the years. However, the goal of the laboratories taking part in these annual exercises is not only to check the accuracy of their results but also to have analytical discussions and the opportunity to exchange experiences that will enrich the group's general competence. PMID- 14527007 TI - Developments in internal monitoring techniques. AB - In an effort to increase accuracy and speed, improve detection limits and reduce uncertainties in internal dosimetry, laboratories have developed improved or new internal monitoring techniques in both in vivo measurements and bioassay analyses. Most of these techniques have not yet entered routine monitoring programmes. This paper intends to summarise these new techniques, show their potential improvements compared to the currently employed monitoring routines and discuss the main aspects of the EC-funded IDEA project, which aims at a comprehensive assessment of these techniques and the enhancements necessary to bring them to broader acceptance in the routine monitoring community. PMID- 14527008 TI - The reduction of limits of detection in in vivo counting of low-energy photon emitters by optimising the shape and size of detectors. AB - This paper compares three ways to reduce the detection limits of in vivo measurements by using passive techniques: the use of shielded rooms, the use of underground laboratories and the adjustment of the detector's dimensions to the examined energy. This study indicates that the efficiency of the detector is not the critical parameter for achieving sensitive measurements in the low-energy range and is secondary to optimisation of the background level. However, reduction of the background has a limited impact on the sensitivity of counting due to 40K in the body. This study also shows that the advantage of using deep underground laboratories could be replaced by detectors with properly designed shape and size and used at ground level in normally shielded rooms to reach the necessary limits of detection for in vivo assessment of low-energy photon emitters. PMID- 14527009 TI - Practical system to detect and assess consequences of radioactivity in a wound. AB - Any wound received within the controlled areas of Dounreay could potentially be contaminated with plutonium and/or uranium isotopes. Wounding bypasses the body's natural barriers to foreign objects, such as the skin and the lining of the alimentary canal. Therefore a relatively low intake can potentially result in a high committed effective dose. The likelihood of contamination of a wound with either plutonium or uranium is low. The decision-making process for dealing with wounds has been continually reviewed and updated by the approved dosimetry service (ADS) at Dounreay. Each wound is considered on an individual basis by the ADS in order to decide what follow-up actions, if any, are necessary. The aim of the process is to maximise the probability that a significant intake is detected, while targeting resources in an appropriate manner. PMID- 14527010 TI - Application of a Ge semi-conductor detector to whole-body counter. AB - To calibrate a whole-body counter, it is necessary to find a determination method for peak efficiencies of detectors used in the whole-body counter. For this purpose, peak efficiencies of a Ge semiconductor detector for point and volume sources were evaluated in the photon energy range 60-1836 keV by Monte Carlo simulation and experiment. It was found that the calculated peak efficiency curves as a function of energy without modelling of the actual sensitive region of the detector are similar in shape to those measured. The calculated peak efficiencies of the detector having an apparent dead layer (1 mm) were also found to agree with the experimental values (deviations from -10-24%). Consequently, the simulation method for peak efficiencies was validated. In addition, an optimum design for a whole-body counter with Ge semiconductor detectors was examined by simulation. This simulation provides a method to determine an optimum arrangement of detectors in a whole-body counter, offering a uniform response to various 137Cs distributions in a human body. PMID- 14527011 TI - A feasibility study for transportable 241Am-in-lung and 241Am-in-nose-blow monitoring systems for use following a weapons accident. AB - In a nuclear weapon accident involving fire or conventional explosion, most of the radiation dose received by people in the immediate vicinity would result from inhalation of 239Pu. This is accompanied by the nuclide 241Am, which is much easier to determine by external counting because of the 60 keV gamma ray emission. In the event of an accident, a priority would be to identify any people who have had intakes of 239Pu which were so large that decorporation therapy should be considered. Direct measurement of lung content provides the most rapid and convenient method for assessing intakes by inhalation. A transportable system has been considered as this could be deployed close to the site of the accident and would allow rapid measurements to be made. The feasibility of a transportable 241Am-in-nose-blow and nasal swab measurement system has also been considered. This would be used to help select people for 241Am-in-lung measurements. PMID- 14527012 TI - Sensitivity of a low energy Ge detector system for in vivo monitoring in the framework of ICRP 78 applications. AB - In in vivo detection of internal contamination by actinides the minimum detectable activities (MDAs) correspond to significant doses, so the sensitivity of the detection system is the key to establishing adequate individual monitoring programmes for internal exposure to these radionuclides. The whole body counting (WBC) faculty at CIEMAT uses a low-energy Ge detector system with different available counting geometries to estimate the retention of actinides in the lungs and evaluate 125I in thyroid and 241Am in bone (skull and knee). A study of the factors and uncertainties involved in estimations of MDA is presented for lung and thyroid monitoring. The dependence of detection limits on counting efficiency in the measurement of low-energy emitters in the lungs has been carefully studied, carrying out a comparison among different biometric equations obtained by ultrasound techniques for estimations of chest wall thickness. Dosimetric implications of the estimated MDAs are taken into account in the framework of ICRP 78 application and considering Spanish regulations. The main interest in lung measurements is for the assessment of occupational exposure. This work confirms the low-energy Ge detector system to be an adequate in vivo technique for the routine monitoring of internal exposure to most insoluble uranium compounds (detection of 3% enriched uranium in lungs), and also to be useful in special monitoring programmes or in the case of incidents when the detection of 241Am is required. PMID- 14527013 TI - Performance of IRD-WBC HPGe detection system for low energy photon emitters in lungs. AB - The Whole Body Counter Facility (WBC) of IRD-CNEN in Brazil has been operating since 1986. The first system installed to perform in vivo measurements of low energy photon emitters radionuclides used Phoswich detectors. In 1998, the WBC unit was upgraded by the installation of an array of four low energy high purity germanium detectors. The performance and suitability of the detection system for lung measurements were evaluated by comparison with the annual dose limits and the detection limits obtained for 238U, 235U, 226Ra and 241Am. This evaluation determined whether the in vivo measurements are adequate. In order to compare the dose limit of 20 mSv y(-1), recommended by the International Commission on Radiological Protection (ICRP), with the in vivo monitoring technique, the minimum detectable intake (MDI) was calculated using the appropriate biokinetic models described in the ICRP Publications. The results were obtained for a single intake through inhalation. The AMAD considered was 5 microm. PMID- 14527014 TI - In vivo measurement technique of 40K as an indicator of the amount of skeletal muscle tissue in the human body. AB - Potassium is an essential element for human metabolism. It is present in all living cells, predominantly in the skeletal muscle tissue. The energy of the 40K photon and its uniform distribution within the human body allow its in vivo measurement. Subjects of both sexes were monitored at the whole-body counter of the IRD facility for the evaluation of 40K body burden, being divided into two groups: (1) subjects who do not exercise routinely, and (2) subjects who do exercise routinely. The average values found for potassium mass and potassium concentration in group 1 were 99 +/- 17 g of K and 1.3 +/- 0.2 g of K kg(-1) of body mass, respectively, and in group 2 the average values found for potassium mass and potassium concentration were 118 +/- 33 g of K and 1.6 +/- 0.2 g of K kg(-1) of body mass, respectively. The comparison between average values for potassium mass and concentration shows a significant statistical difference. PMID- 14527015 TI - Current status of internal dosimetry in Lithuania. AB - After Lithuania regained independence, the legal basis for existing radiation protection was modified radically according to the IAEA, ICRP recommendations and the requirements of legislation of the European Community. The legal basis for internal dosimetry and a functioning system of assessment of exposure to intake of radionuclides have been created in the Radiation Protection Centre (regulatory authority in radiation protection). Direct and indirect measurements of concentrations of radionuclides are used for the assessment of internal doses of workers and the public. PMID- 14527016 TI - The medical management of unintentional radionuclide intakes. AB - As a general medical problem, radionuclide intakes that may cause significant health effects are uncommon events. In preparing to manage a radionuclide accident, planning is the key. The medical aspects of such an accident are only one part of the management, and a professional team approach is required. Specific priorities and sequencing are necessary in medically managing a radionuclide intake. As soon as is reasonably practical, promptly remove the victim(s) from further radionuclide, radiation field, or chemical exposure. Life and limb-saving medical aid takes precedence over ionising radiation concerns in nearly all cases. Next are the prevention and/or minimisation of internal intake of radionuclides and evaluation and control of external radionuclide contamination, followed by institution of treatment to minimise the retained radionuclide. Communication with the accident victim, and his or her family, and public affairs/media issues are important. Finally, follow-up treatment for internal intakes that may cause delayed health effects is given. PMID- 14527017 TI - Successful DTPA therapy in the case of 239Pu penetration via injured skin exposed to nitric acid. AB - This paper presents results of the radiological study and DTPA therapy for a worker exposed to a plutonium nitrate solution. Plutonium levels were measured in excreta, blood, plasma and wound for several weeks. Plutonium renal clearance ranged from 110-190 ml min(-1) to 3-4 ml min(-1) at different stages of chelation therapy. Plutonium absorption into blood from the injured skin amounted to 4.3%. As a result of intensive therapy, 96% of absorbed plutonium was successfully excreted. PMID- 14527018 TI - Octadentate hydroxypyridinonate (HOPO) ligands for plutonium (i.v.): pharmacokinetics and oral efficacy. AB - Linear octadentate spermine based 3,4,3-LI(1,2-HOPO) and the mixed ligand, 3,4,3 LI(1,2-Me-3,2-HOPO), are the most effective agents for decorporation of Pu prepared so far; they are effective at low dosage, orally active, and of low toxicity at effective injected dosage. Their pharmacological properties are favourable for in vivo Pu chelation--penetration of extracellular water, useful residence in the circulation, substantial hepato-biliary excretion, low but useful GI absorption, and transitory residence in the kidneys. Reductions of body Pu were significant, compared with controls, when oral administration to normally fed mice (30 or 100 micromol kg(-1)) was delayed as long as 24 h after i.v. Pu injection. The HOPO ligands (10-100 micromol kg(-1)) or CaNa3-DTPA (100 or 300 micromol kg(-1)) were given orally to normally fed mice starting at 4 h after an i.v. Pu injection and continued 5 d per week for 3 weeks. 3,4,3-LI(1,2-HOPO) (100 micromol kg(-1)) reduced Pu in skeleton, liver, and body, to 44 +/- 9, 18 +/- 8, and 38 +/- 7% of controls, respectively, reductions significantly greater than with the mixed HOPO ligand or with three times more CaNa3-DTPA. PMID- 14527019 TI - An analysis of a puncture wound case with medical intervention. AB - A worker noted a small wound to his thumb when leaving a work site that was undergoing decontamination because of past operations with plutonium (Pu) and americium (Am). Direct surveys of the wound site confirmed the presence of contamination. The chelating agent Ca-DTPA was administered via a nebuliser within an hour after discovery of the wound. External measurements were made of the wound site and wound dressings; 24-h urinary excretion data were collected periodically and the Pu and Am urine content was determined. Zn-DTPA was administered on three occasions. The ICRP Pu systemic model was modified to consider the enhanced urinary excretion following administration of the chelating agents. The analysis indicated that the wound resulted in an initial deposition of 400 Bq 238Pu, 2240 Bq (239/240)Pu and 1060 Bq 241Am. About 70% of the initial wound activity was removed by surgical procedures and less than 1% of the wound activity was removed by chelation therapy. This paper compares the observed urinary excretion data with that indicated by a simulation of the kinetics of the transfer from the wound site and the kinetics of the chelating agent and Pu. PMID- 14527020 TI - Improved chelation therapy of intramuscularly deposited thorium by CaDTPA in the rat. AB - Comparative studies on the translocation and retention of intramuscularly (i.m.) injected thorium nitrate (234Th 46 ng + 232Th 5 microg per rat) in solutions of citrate, CaDTPA or citrate + CaDTPA in rats have been conducted. Results showed that only thorium in mixed-ligand solution was entirely translocated from the muscle, with the greatest part being excreted from the body. In this case, the whole-body retention of thorium decreased to 16% of the injected radioactivity within 2 d, 13% being retained in the skeleton. Studies on the decorporation of 234Th + 232Th nitrates from a rat wound simulated with i.m. injection have also been carried out. The greatest translocation of thorium and its excretion was achieved with a single local injection of the mixed-ligand (citrate + CaDTPA) solution when compared with those of citrate or CaDTPA alone. The efficiency of mixed-ligand treatment decreased with its delay. On day 2 post-therapy, the whole body content of thorium decreased to 30, 37 and 55% of injected radioactivity when the local treatment started immediately, postponed to 1 h or 24 h, after i.m. injection of thorium, respectively. In control rats without treatment, there was only a slight decrease in the content of thorium in the whole body. PMID- 14527021 TI - Effect of DTPA on the nephrotoxicity induced by uranium in the rat. AB - The only treatment proposed after human contamination with MOX (mixed oxide of uranium and plutonium) is diethylenetriaminepentaacetic acid (DTPA), because plutonium is considered to be the major risk. However, both DTPA and uranium are nephrotoxic at high dosages and DTPA has been shown to increase in vitro the cytotoxicity induced by uranium on cultured epithelial tubular cells. This work aimed to test this effect in vivo. Rats were injected with subtoxic (57 microg kg(-1)) to toxic (639 microg kg(-1)) amounts of uranium as nitrate at 0 h, they received two DTPA injections (30 micromol kg(-1)) at 2 min and 24 h and were euthanased at 48 h. The nephrotoxic effects were evaluated by measurement of the body weight gain, food and water intake, measurement of biochemical parameters in urine and blood, and histological examination of one kidney. The main result was that DTPA did not increase the nephrotoxicity induced by uranium in the range of concentrations tested, which was inconsistent with the in vitro results. PMID- 14527022 TI - Competitive binding of Pu and Am with bone mineral and novel chelating agents. AB - Effective direct removal of actinides such as Pu and Am from bone in vivo has not been accomplished to date, even with the strong chelating agents CaNa3DTPA or ZnNa3DTPA. This study, using an established in vitro system, compared removal of Pu and Am bound to bone mineral by ZnNa3DTPA and 10 chelating agents designed specifically to sequester actinides, including Pu and Am. Ligands tested were tetra, hexa, and octadentate, with linear or branched backbones containing sulfocatechol [CAM(S)], hydroxycatechol [CAM(C)], hydroxipyridinone (1,2-HOPO, Me 3,2-HOPO), or hydroxamate functional groups. The wide range of Pu and Am removal exhibited by the test ligands generally agreed with their metal coordination and chemical properties. The most effective agents for Pu (100 microM concentration, 24-48 h contact) are all octadentate as follows: 3,4,3-LICAM(S) (54% unbound); 3,4,3-LICAM(C) (6.2%); 3,4,3-LI(1,2-HOPO) (3.8%); H(2,2)-(Me-3,2-HOPO) (2.2%) and DFO-(1,2-HOPO) (1.8%). The other ligands removed less than 1% of the bound Pu; and ZnNa3DTPA removed only 0.086%. The most effective ligands for Am removal (100 microM, 24-48 h contact) are as follows: octadentate H(2,2)-(Me-3,2-HOPO) (21% unbound); 3,4,3-LI(1,2-HOPO) (14.5%) and 3,4,3-LICAM(C) (5.9%); hexadentate TREN (Me-3,2-HOPO) and TREN-(1,2-HOPO) (9.6%); and tetradentate 5-LIO(Me-3,2-HOPO) (5.2%). Am removal by ZnNa3DTPA was about 1.4%. Among the ligands presently considered for possible human use, only 3,4,3-LI(1,2-HOPO) removed potentially useful amounts of both Pu and Am from bone mineral. PMID- 14527023 TI - Comparative decorporation efficacy of 3,4,3-LIHOPO, 4,4,4-LIHOPO and DTPA after contamination of rats with soluble forms of 238Pu and 233U. AB - The aim of this study was to compare the efficacies of DTPA, 3,4,3-LIHOPO and a newly synthesised molecule, 4,4,4-LIHOPO, in removing 233U and 238Pu after internal contamination by soluble forms of those nuclides. For this purpose, intravenous injections of DTPA (30 micromol kg(-1)) or 3,4,3-LIHOPO or 4,4,4 LIHOPO at dosages of 0.3 or 30 micromol kg(-1) were performed 1, 6 and 24 h after contamination of rats by intravenously injected 238Pu citrate and 1 h after intravenous injection of 233U nitrate. Actinide content in the main retention organs and cumulated excretion were measured 48 h after contamination. These experiments show similar decorporation efficacies of 4,4,4-LIHOPO and 3,4,3 LIHOPO for Pu, which are much higher than that of DTPA. At a dosage of 0.3 micromol kg(-1), the two LIHOPO analogues were as efficient as DTPA at a dosage of 30 micromol kg(-1). After U contamination, a 20% decorporation efficacy was obtained for either 3,4,3-LIHOPO or 4,4,4-LIHOPO at a dosage of 30 micromol kg( 1). PMID- 14527024 TI - The application of voxel phantoms to the internal dosimetry of radionuclides. AB - Extensive calculations of specific absorbed fractions (SAFs) for monoenergetic photon sources were performed using a Monte Carlo photon transport code together with seven male and female adult voxel models based on computed tomographic data of real persons. These models offer greater realism with respect to organ topology than the mathematical phantoms commonly used in the past. Due to individual anatomical differences, large variations in photon SAFs between the voxel models were found that can amount to orders of magnitude for very low photon energies. However, in many cases, the larger differences were found between MIRD-type and voxel models, since the inter-organ distances tend to be larger in the MIRD-type phantoms than in reality, due to over-simplification of organ shapes. Furthermore, organ absorbed doses per incorporated activity were evaluated for two selected radiopharmaceuticals. Although a method was found to largely eliminate the influence of organ mass on SAFs for organ self-absorption, the absorbed dose coefficients varied by several tens of per cent between the individual voxel models, thus indicating a significant influence of individual photon SAFs for organ cross-fire on organ absorbed dose. Again, 43% of the MIRD organ dose values were outside the range of doses spanned by the voxel models. Effective dose showed a variation of only up to 26% between the single voxel models for the radiopharmaceuticals considered. PMID- 14527025 TI - Voxel phantoms and Monte Carlo methods applied to in vivo measurements for simultaneous 241Am contamination in four body regions. AB - A Monte Carlo program, Visual Monte Carlo (VMC) in vivo, was written to simulate photon transport through an anthropomorphic phantom and to detect radiation emitted from the phantom. VMC in vivo uses a voxel phantom provided by Yale University and may be used to calibrate in vivo systems. This paper shows the application of VMC in vivo to the measurement of 241Am deposited simultaneously in the thoracic region, the bones, the liver and in the rest of the body. The percentages of 241Am in the four body regions were calculated using the biokinetic models established by the ICRP, for a single intake via inhalation. The four regions of the voxel phantom were then 'contaminated' in accordance with the calculated percentages. The calibration factor of the in vivo system was then obtained. This procedure was repeated for the radionuclide distributions obtained 5, 30, 120, 240 and 360 days after intake. VMC in vivo was also used to calculate the calibration factor of the in vivo system in which the radionuclide was assumed to be deposited only in the lung, as is normally done. The activities calculated with the radionuclide distributed in the four body regions as a factor of time, and the activities calculated with the radionuclide deposited in the lung only are compared. PMID- 14527026 TI - Monte Carlo simulations: a useful tool to extend in vivo calibrations and explore alternative approaches. AB - Designing and testing new equipment can be an expensive and time consuming process or the desired performance characteristics may preclude its construction due to technological shortcomings. Cost may also prevent other types of scenario being tested. An alternative is to use Monte Carlo simulations to make the investigations. This paper exemplifies how Monte Carlo code calculations can be used to fill the gap by describing two investigations: (1) the possible self attenuation of homogeneously distributed natural uranium in a lung phantom; and (2) the effect of activity deposited in the ribs on the activity estimate from a lung count. PMID- 14527027 TI - Evaluation of specific absorbed fractions in voxel phantoms using Monte Carlo simulation. AB - There is a need to calculate specific absorbed fractions (SAFs) in voxel phantoms for internal dosimetry. For this purpose, an EGS4 user code for calculating SAFs using voxel phantoms was developed on the basis of an existing EGS4 user code for external dosimetry (UCPIXEL). In the developed code, the transport of photons, electrons and positrons in voxel phantoms can be simulated, particularly the transport simulations of secondary electrons in voxel phantoms. The evaluated SAFs for the GSF 'Child' voxel phantom using the developed code were found to be in good agreement with the GSF evaluated data. In addition, SAFs in adult voxel phantoms developed at JAERI were evaluated using the developed code and were compared with several published data. It was found that SAFs for organ self absorption depend on the organ masses and would be affected by differences in the structure of the human body. PMID- 14527028 TI - Development and assessment of a new radioactive decay database used for dosimetry calculation. AB - The present paper discusses the technical issues and a scheme for their resolution before undertaking the development of a new radioactive decay database to succeed ICRP38. The scheme consists of the following procedures. (1) The consistency of nuclear structure data files used as input to the computer code EDISTR is established by referring to the latest nuclear parameters and by comparing the computed energies of emitted radiations with total decay energy. (2) A method for calculating detailed spectra of X rays and Auger electrons is incorported into EDISTR to enhance the treatment of atomic relaxation processes initiated by electron vacancies from internal conversion and electron capture. (3) Quality assurance of the compiled data is undertaken by comparisons with experimental data and other decay databases prepared from different computer codes and libraries. JAERI and ORNL are cooperating to implement the scheme in order to assemble a comprehensive radioactive decay database including over 1000 nuclides. PMID- 14527029 TI - Reference values for basic human anatomical and physiological characteristics for use in radiation protection. AB - A new publication prepared by the ICRP Task Group on Reference Man, Basic anatomical and physiological data for use in radiological protection: reference values, is focused on those human characteristics that are important for dosimetric calculations. Moving from the past emphasis on a Reference Man, the new report presents a series of reference values for both male and female subjects of six different ages--newborn, 1, 5, 10, 15 y, and adult. In selecting reference values, the task group has used data on Western Europeans and North Americans because these populations have been well studied with respect to anatomy, body composition and physiology. When appropriate, comparisons are made between the chosen reference values and data from several Asian populations. The reference values for height and body mass are higher than those reported for various Asian populations. However, the reported masses of individual organs and tissues, particularly for China and Japan, are similar to the reference values. PMID- 14527030 TI - Developments in the internal dosimetry of radiopharmaceuticals. AB - Various radionuclides are used in nuclear medicine in different diagnostic and therapeutic procedures. Recently, interest has grown in therapeutic agents for some interesting applications in nuclear medicine. Internal dose models and methods in use for many years are well established, and can give radiation doses to stylised models representing reference individuals. Kinetic analyses need to be carefully planned, and dose conversion factors that are most similar to the subject in question should be chosen, which can then be tailored somewhat to be more patient-specific. Internal dose calculations, however, are currently not relevant in patient management in internal emitter therapy, as they are not sufficiently accurate or detailed to guide clinical decision-making, and as calculated doses have historically not been well correlated with observed effects on tissues. Great strides are being made at many centres regarding the use of patient image data to construct individualised voxel-based models for more detailed and patient-specific dose calculations, and new findings are encouraging regarding improvement of internal dose models to provide better correlations of dose and effect. These recent advances make it likely that the relevance will soon change to be more similar to that of external beam treatment planning. PMID- 14527031 TI - Assessments for high dose radionuclide therapy treatment planning. AB - Advances in the biotechnology of cell specific targeting of cancer and the increased number of clinical trials involving treatment of cancer patients with radiolabelled antibodies, peptides and similar delivery vehicles have led to an increase in the number of high dose radionuclide therapy procedures. Optimised radionuclide therapy for cancer treatment is based on the concept of absorbed dose to the dose limiting normal organ or tissue. The limiting normal tissue is often the red marrow, but it may sometimes be the lungs, liver, intestinal tract or kidneys. Appropriate treatment planning requires assessment of radiation dose to several internal organs and tissues, and usually involves biodistribution studies in the patient using a tracer amount of radionuclide bound to the targeting agent and imaged at sequential timepoints using a planar gamma camera. Time-activity curves are developed from the imaging data for the major organ tissues of concern, for the whole body and sometimes for selected tumours. Patient specific factors often require that dose estimates be customised for each patient. In the United States, the Food and Drug Administration regulates the experimental use of investigational new drugs and requires 'reasonable calculation of radiation absorbed dose to the whole body and to critical organs' using the methods prescribed by the Medical Internal Radiation Dose (MIRD) Committee of the Society of Nuclear Medicine. Review of high dose studies shows that some are conducted with minimal dosimetry, that the marrow dose is difficult to establish and is subject to large uncertainties. Despite the general availability of software, internal dosimetry methods often seem to be inconsistent from one clinical centre to another. PMID- 14527032 TI - A generic model for 11C labelled radiopharmaceuticals for imaging receptors in the human brain. AB - A large number of radiopharmaceuticals labelled with 11C (half-time 0.340 h) are being developed for positron emission tomographic studies of different types of receptor in the human brain. For most of these agents, the available biokinetic data are insufficient to construct realistic compound-specific biokinetic models for calculating the internal radiation dose delivered to persons undergoing investigation. A generic model for brain receptor substances that predicts the internal dose with sufficient accuracy for general radiation protection purposes has, therefore, been developed. Biokinetic data for 13 11C-radiopharmaceuticals used clinically for imaging different brain receptors indicate that, despite differences in chemical structure, their uptake and retention in the human brain and other tissues are broadly similar. The proposed model assumes instantaneous deposition of 5% of the injected radioactivity in the brain, with the remaining radioactivity being rapidly and uniformly distributed throughout all other tissues. Elimination from all tissues is assumed to occur with a half-time of 2 h. It is further assumed that 75% of the injected 11C is excreted in the urine, and 25% via the gall bladder, with a half-time of 2 h. This model yields an effective dose of 4.5 x 10(-3) mSv MBq(-1), with doses of 3.2 x 10(-2), 1.7 x 10( 2), 8.7 x 10(-3), 5.2 x 10(-3), and 3.8 x 10(-3) mGy MBq(-1) to the urinary bladder, gall bladder, kidneys, brain and ovaries, respectively. These doses are well within the range of those reported using compound-specific models for the radiopharmaceutals studied. PMID- 14527033 TI - Contributions of short-lived radioiodines to thyroid doses received by evacuees from the Chernobyl area estimated using early in vivo activity measurements. AB - A series of in vivo gamma spectrometric measurements of 65 people evacuated from Pripyat 1.5 days after the Chernobyl Nuclear Power Plant Unit 4 explosion was performed in St Petersburg, Russia, as early as 30 April 1986. The historical spectra and interviews were recently processed and the results used for thyroid dose estimation. Activities of 131I in thyroid and 132Te in lungs were determined easily; for estimation of 132I and 133I activities in thyroid, sophisticated methods of spectral processing were developed. According to thyroid measurement data, the mean ratio of 133I/131I activities (at the time of the accident) inhaled by residents of Pripyat was 2.0. The mean ratio of thyroid dose from 133I inhalation to that caused by 131I amounts to 0.3, which confirms the accuracy of dose estimates based on the evolution of the Chernobyl accident. The mean ratio of 132I activity in thyroid to that of 132Te in lungs was assessed from the human measurement data to be 0.2, which is in reasonable agreement with the metabolic properties of these radionuclides. The mean ratio of thyroid dose from 132I originating from 132Te deposited in lungs to the dose caused by 131I was 0.13 +/- 0.02 for Pripyat residents who did not take KI pills and 0.9 +/- 0.1 for persons who took KI pills. Thus, the contribution of short-lived radioiodines to total thyroid dose of Pripyat residents, which was on average 30% for persons who did not use stable iodine prophylaxis, and about 50% for persons who took KI pills on 26-27 April, should be accounted for in the assessment of thyroid health effects. PMID- 14527035 TI - Assessment of doses to the offspring of the Techa River cohort due to intakes of radionuclides by the mother. AB - The Techa River was contaminated as a result of radioactive releases by the Mayak plutonium production facility in 1949-1956. The residents of riverside communities were exposed to internal irradiation from radionuclides ingested mainly with river water, and also to external gamma irradiation resulting from shoreline and flood-plain contamination. The most important role in population exposure was played by (89,90)Sr and 137Cs. The persons born after the onset of the contamination have been identified as the 'Techa River Offspring Cohort' (TROC). The TROC has the potential to provide direct data on health effects in progeny that resulted from exposure of a general population to chronic radiation. This report describes the results of the calculation of fetal doses due to intakes of radionuclides by their mothers. Particular attention has been given to fetal dose from 90Sr because this nuclide is the most significant in terms of population dose for the Techa River. The comparison of the fetal bone marrow doses evaluated using different approaches proposed in the literature has shown a large dispersal in dose values. The main cause of this is the difference in model assumptions simplifying some developmental aspects of fetal haematopoiesis and bone formation. This paper presents an analysis of these basic assumptions that could be useful for further improvements in fetal dosimetry. PMID- 14527034 TI - Uncertainties in thyroid dose reconstruction after Chernobyl. AB - The most significant impact of the Chernobyl accident of 26 April 1986 is the increased incidence of thyroid cancer among Belarusians, Russians and Ukrainians who were exposed as children to radioiodines in fallout resulting from atmospheric releases. The US National Cancer Institute (NCI), in cooperation with the ministries of health of Belarus and Ukraine, is involved in epidemiological studies of thyroid diseases related to the accident. Individual thyroid doses, as well as uncertainties, have been estimated for the members of the cohort studies (approximately 13,000 Ukrainians and 12,000 Belarusians). The cohort subjects, who were selected from the large group of children whose thyroids were monitored for gamma radiation within a few weeks of the accident, provided personal information on their residence history and dietary habits during interviews. Thyroid dose estimates range from 1 mGy to more than 20 Gy. The uncertainties are found to be approximately log-normally distributed, with geometric standard deviations ranging from 1.6 to 5.0. The medians of the geometric standard deviations are 1.7 for the Ukrainian subjects and 2.1 for the Belarusian subjects. The major sources of uncertainty in the thyroid dose estimates are found to be those related to the thyroid mass of the subject and to the estimation of the thyroidal content of 131I at the time of thyroid monitoring. PMID- 14527036 TI - Estimation of human exposure to natural radionuclides using in vivo skull measurements. AB - In a preliminary study, in vivo skull measurements and in vitro urine measurements of 210Pb and nulU have been performed to find out the individual, chronic exposure to waterborne natural radionuclides of a small group of Finnish people. For their domestic water, the studied individuals use water from drilled wells containing elevated concentrations of natural uranium and its daughter nuclides ((234,235,238)U, 222Rn, (226,228)Ra, 210Po, 210Pb). Enhanced 210Pb and 235U activities were observed in several people. A positive correlation is observed between the U concentration in urine (microg d(-1)) and the number of counts (cpm) in the gamma ray energy peaks originating from the decay of 235U and 234Th respectively. Calibration of the detector set-up and the determination of background sources are in progress. PMID- 14527037 TI - Improvements in the biokinetic model for strontium with allowance for age and gender differences in bone mineral metabolism. AB - An age- and gender-dependent biokinetic model for strontium was developed based on the study of a population living along the Techa River exposed to effluents from the Mayak Production Association (MPA). To estimate the parameters of a new model (Techa biokinetic model, TBM) many data sets have been assembled: our whole body counter data on long-term retention of 90Sr in humans, data from studies during the period of global fallout, data resulting from deliberate injection of strontium radionuclides and non-radiological data regarding bone formation and resorption, mineral content of the body, etc. The model was developed using the basic structure of the ICRP biokinetic model for strontium, but new age- and gender-specific parameters were derived. This paper discusses the approaches applied to develop the new model. PMID- 14527038 TI - 131I content in the human thyroid estimated from direct measurements of the inhabitants of Russian areas contaminated due to the Chernobyl accident. AB - The method of processing and the results of measurements of 131I content in the thyroids of Russian people performed in May-June 1986 are presented. The contribution of radiation from Cs radionuclides in the human body was taken into account in the processing of measurement data with an SRP-68-01 device. The greatest individual 131I content was found in the thyroids of inhabitants of the Bryansk region, up to 250-350 kBq, and in the Tula and Orel regions, up to 100 kBq. The average 131I thyroid activity in the middle of May 1986 reached 80 kBq for inhabitants of some settlements in the Bryansk region, 5-8 kBq in the Tula region and 5 kBq in the Orel region. PMID- 14527039 TI - RBDATA-EULEP: providing information to improve internal dosimetry. AB - The overall aim of the concerted action RBDATA-EULEP is to provide information to improve the assessments of intakes of radionuclides and of the resulting doses. This involves a review of the behaviour of radionuclides following intake, and the transfer of expertise on methodology by organising small training workshops. The main activity is the development of an electronic database, effectively an annotated bibliography, but the electronic format used facilitates extension, updating and information retrieval. It consists of linked tables of references and experiments, with details and comments on the materials, procedures and results. By June 2002 it contained information on 524 inhalation, 282 ingestion and 164 injection experiments from 391 references. It will be extended, and Internet access provided. Prospective users include groups developing standards for internal dosimetry, scientists conducting research on radionuclide biokinetics and health physicists assessing the consequences of accidental intakes. PMID- 14527040 TI - Biokinetics of radionuclides and treatment of accidental intakes. AB - This paper describes the objectives and reviews the progress of EULEP Working Party 5, convened under the auspices of the European Union's Fifth Framework Programme, to 'cluster' two EU-supported contracts, Biokinetics and Dosimetry of Internal Contamination (BIODOS (EU Contract FIS5-1999-00214)) and Radionuclide Biokinetics Database (EULEP) (RBDATA-EULEP (Concerted Action Contract FIS5-1999 00218), and two non-EU funded projects, Biokinetics of Radionuclides in Human Volunteers (RNHV (non-EU funded project)) and Treatment of Accidental Intakes of Radionuclides (TAIR (part funded by EULEP)). PMID- 14527041 TI - Design and implementation of monitoring programmes for internal exposure (project OMINEX). AB - The costs of monitoring for internal exposure in the workplace are usually significantly greater than the equivalent costs for external exposure. Therefore, there is a need to ensure that resources are employed with maximum effectiveness. The EC-funded OMINEX (optimisation of monitoring for internal exposure) project is developing methods for optimising the design and implementation of internal exposure monitoring programmes. Current monitoring programmes are being critically reviewed, the major sources of uncertainty in assessed internal dose investigated, and guidance formulated on factors such as programme design, choice of method/techniques, monitoring intervals, and monitoring frequency. OMINEX will promote a common, harmonised approach to the design and implementation of internal dose monitoring programmes throughout the EU. PMID- 14527042 TI - Guidance on internal dose assessments from monitoring data (project IDEAS). AB - Several international inter-comparison exercises on intake and internal dose assessments from monitoring data led to the conclusion that the results calculated by different participants varied significantly, mainly due to the broad variety of methods and assumptions applied in the assessment procedure. Based on these experiences, the need of harmonisation of the procedures has been formulated as an EU research project under the 5th Framework Programme, with the aim of developing general guidelines for standardising assessments of intakes and internal doses. In the IDEAS project, eight institutions from seven European countries are participating, also using inputs from internal dosimetry professionals from across Europe to ensure broad consensus in the outcome of the project. To ensure that the guidelines are applicable to a wide range of practical situations, the first step will be to compile a database on well documented cases of internal contamination. In parallel, an improved version of existing software will be developed and distributed to the partners for further use. Many cases from the database will be evaluated independently by more partners using the same software and the results will be discussed and the draft guidelines prepared. The guidelines will then be revised and refined on the basis of the experiences and discussions of two workshops, and an intercomparison exercise organised in the frame of the project which will be open to all internal dosimetry professionals. PMID- 14527043 TI - Internal dosimetry: enhancements in application, update on the IDEA project. AB - Efforts in many internal dosimetry laboratories to increase the accuracy and speed of measurements, to improve detection limits and to reduce uncertainties have resulted in improved or new internal monitoring techniques, both in in vivo measurement and in bioassay analysis. The EC-funded IDEA project (internal dosimetry--enhancements in application) aims to investigate why most of these techniques have not yet entered routine monitoring programmes (a summary of these new techniques is given here, showing their potential improvements compared with the currently employed monitoring routines). The project further aims at a comprehensive assessment of these techniques and the enhancements necessary to bring them to broader acceptance with those performing routine monitoring. PMID- 14527044 TI - Harmonisation (legal, dosimetric, quality aspects) of individual monitoring, and integration of monitoring for external and internal exposures (EURADOS Working Group). AB - The EURADOS Working Group II on 'Harmonisation of individual monitoring' consists of experts from almost all EU Member States and Newly Associated States (NAS), involved in tasks related to the assessment of doses for internal and external radiation. The final objective is to achieve harmonisation in individual monitoring for occupational exposures. Sub-group 2 activities are focused on investigating how the results from personal dosemeters for external radiation and workplace monitoring and from monitoring for internal exposure can be combined into a complete and consistent system of individual monitoring. Three questionnaires were prepared, covering 'Individual monitoring of external radiation' (Questionnaire 1), 'Internal exposure' (Questionnaire 2) and 'Natural sources of radiation at the workplace' (Questionnaire 3). With the agreement of a 'contact-person', selected in each country, the distribution of the three EURADOS 2002 questionnaires was carried out by e-mail among the dosimetry facilities of 28 European countries. The preliminary results of these actions are presented here. PMID- 14527046 TI - Breast cancer knowledge, beliefs, and screening behaviors among low-income, elderly black women. AB - This cross-sectional study explored whether there are age-specific differences in breast cancer-related knowledge, beliefs, and screening behaviors among low income, elderly black women. Data were collected at senior citizen facilities from 214 black women aged 65 and older. Differences in knowledge, beliefs, and screening practices across three age groups were assessed by chi-square tests. Logistic regression modeling was used to determine the effect of these factors on compliance with American Cancer Society (ACS) screening guidelines. Age was inversely associated with knowledge and screening practices. The youngest group (65-74) was about twice as likely as the oldest group (85 and over) to correctly recognize breast cancer risk factors. About 50% of the oldest women compared to about 20% of the youngest women believed their risk for breast cancer was nil. The oldest group was also least likely to have had a mammogram or clinical breast examination within the past year, as recommended by the ACS. Our results suggest that educating elderly women, especially those 85 and over, about breast cancer and screening may lead to higher compliance with ACS recommendations. PMID- 14527045 TI - Ethnic disparities in type 2 diabetes: pathophysiology and implications for prevention and management. AB - Based on 2000 census data, ethnic minorities constitute approximately 25% of the overall population of the United States, and the population of minority groups has been increasing at a faster rate than the general U.S. population. Compared with caucasians, persons from minority ethnic groups suffer disproportionately from type 2 diabetes and its long-term complications. Acute complications of diabetes occur with varying frequencies in the different demographic groups, but there are indications that the rate of hospitalization for diabetic ketoacidosis and nonketotic coma may be higher among certain minority populations. Genetic and lifestyle factors likely account for the increased prevalence of type 2 diabetes among ethnic minorities. However, the increase in morbidity and mortality from diabetes may be the result, in part, of socioeconomic factors. Pathophysiologically, several studies have documented a higher prevalence of insulin resistance in minority groups, even after correction for obesity and lifestyle factors. These findings underscore the need for a more aggressive approach to diabetes management in high-risk populations. Behavioral and pharmacologic interventions that reduce insulin resistance have profound beneficial effects in African-American patients and subjects with diabetes from other ethnic groups. Indeed, much of the ethnic difference in morbidity from diabetic complications disappears when caucasians and non-caucasians are treated to identical degrees of glycemic control. PMID- 14527047 TI - Colorectal cancer screening among African Americans: the importance of physician recommendation. AB - INTRODUCTION: African Americans have higher colorectal cancer incidence and mortality rates than whites. They are also more likely to be diagnosed with late stage disease and less likely to survive for at least five years after diagnosis. Lack of adherence to colorectal cancer screening recommendations has previously been found to be associated with lower income, lower educational level, and racial/ethnic minority status. METHODS: One hundred-fifty African-American patients (aged 50-79 years) of an inner city hospital, were surveyed by mail and telephone in early 2002. Seventy-six patients completed the survey, and data from 74 surveys were analyzed. RESULTS: Approximately one-half (55%) of the respondents reported having received a fecal occult blood test (FOBT) in the last 12 months, sigmoidoscopy in the last five years, or colonoscopy in the last 10 years. Thirty-nine percent of the survey participants reported never having received a physician recommendation for FOBT, 60% reported never having received a recommendation for sigmoidoscopy, and 57% reported never having received a recommendation for colonoscopy. Previous physician recommendation was strongly associated (p < 0.001) with levels of FOBT, sigmoidoscopy, and colonoscopy use. DISCUSSION: Future studies should examine factors that influence primary care physicians' decision-making about ordering colorectal cancer screening tests, as well as patients' decision-making regarding adherence to physician recommendations. PMID- 14527048 TI - Reduced cholesterol levels in African-American adults with sickle cell disease. AB - In a recent study of boys and girls with sickle cell disease (SCD) in Nigeria, a common finding with this genetic hematologic disorder was a marked reduction in total cholesterol. Epidemiologic studies have identified a relation between low serum levels of total cholesterol (< 130 mg/dL) and increased mortality from all causes. We were interested in knowing if hypocholesterolemia was present in African-American adults with SCD. We therefore compared the plasma lipid profiles of the 16 men and 20 women with SCD who received care at the University of Texas Medical Branch at Galveston between 1996 and 2001 with those of 2,415 gender matched African Americans who were seen at the same hospital but who did not have SCD. The age-adjusted mean total cholesterol concentrations of the SCD males and females were 147 +/- 42 mg/dL and 179 +/- 36 mg/dL, compared to male and female control values of 200 +/- 75 mg/dL and 216 +/- 61 mg/dL, respectively. These differences between SCD subjects and controls were statistically significant (p < 0.001). The LDL-cholesterol levels of the men and women with SCD (68 +/- 28 mg/dl and 95 +/- 33 mg/dl, respectively) were also significantly reduced relative to the controls (121 +/- 58 mg/dl and 128 +/- 54 mg/dl, respectively, p = 0.001). The triglyceride levels of the men with SCD were much reduced relative to the male controls (102 +/- 34 mg/dL versus 194 +/- 215 mg/dL, p = 0.02), but were not different between the SCD females and their control group. The HDL-cholesterol levels of the SCD subjects and the controls were not different. These results indicate that total cholesterol and LDL-cholesterol concentrations are significantly reduced in adult men and women with SCD in the United States, and should heighten interest in the implication that low levels of cholesterol might exacerbate the medical problems inherent in this genetic disease. PMID- 14527049 TI - Lower prevalence of intraventricular block in African-American patients compared with Caucasian patients: an electrocardiographic study II. AB - BACKGROUND: Electrocardiographic (ECG) differences occur between African-American and white persons. METHODS: Intraventricular conduction abnormalities of ECGs of 2,123 African-American and white hospital patients ages 20-99 years were studied in a consecutive manner. RESULTS: Intraventricular conduction abnormalities develop later in life and are less common in African-American patients, compared with white patients. The prevalence of conduction abnormalities increases with advancing age in both races. Left- and right ventricular conduction abnormalities begin to rise at age 50 for white patients but begin to rise at age 70 for African-American patients. The prevalence of left ventricular conduction abnormalities peaks in the ninth decade of life in both races but declines in both races in the tenth decade of life. The prevalence of right ventricular conduction abnormalities gradually increases and peaks in the tenth decade of life in both races. CONCLUSIONS: The prevalence of intraventricular block is significantly less in African-American patients, compared with white patients- occurring in 8.6% of African-American patients and in 15.2% of white patients. The prevalence of intraventricular block is lowest in African-American women at 6.5% and highest in white men at 16.8%. PMID- 14527050 TI - Inadequate follow-up for abnormal Pap smears in an urban population. AB - PURPOSE: To determine the factors associated with inadequate follow-up for abnormal Pap smears among a cohort of Boston women from urban academic clinics. METHODS: Subjects were women > 18 years with abnormal cervical cytology between February 1999 and April 2000. Inadequate follow-up was defined as lack of subsequent cervical cytology or pathology specimen within four months of the initial abnormal specimen for high-grade lesions or within 7 months for low-grade lesions. RESULTS: Of the 423 subjects, the mean age was 33 years. Sixty percent were black, 23% Hispanic, 15% white, 2% Asian. The population was largely uninsured or publically insured. The overall inadequate follow-up rate was 38%. In bivariate analysis, age was a significant risk factor; 46% of women ages 18-29 had inadequate follow-up (p < 0.01). In multivariate analysis, women aged 18-29 years were more likely than women 50 years and older to have inadequate follow-up (OR 2.7, 95% CI 1.1-6.4), as were women with Medicaid insurance compared with private insurance (OR 1.9, 95% CI 1.01-3.5). After 12 months, 26% of women with abnormal Pap smears still had not received follow-up. CONCLUSIONS: In a predominantly urban minority population, the overall rate of inadequate follow-up for abnormal Pap smears was high at 38%. Programs to address follow-up of abnormal cervical cytology should focus on minority populations, especially younger and all low-income women. PMID- 14527051 TI - Health care providers' training, perceptions, and practices regarding stress and health outcomes. AB - In order to assess health care providers' training, perceptions, and practices regarding stress and health outcomes, a survey was administered to primary care providers in the outpatient medical clinics of a southeastern urban hospital serving a predominantly African-American indigent population. One-hundred-fifty one of 210 providers (72%) responded. Forty-two percent of respondents reported receiving no instruction regarding stress and health outcomes during their medical/professional education. While 90% believed stress management was "very" or "somewhat" effective in improving health outcomes, 45% "rarely" or "never" discussed stress management with their patients. Respondents were twice as likely to believe that counseling patients about smoking, nutrition, or exercise was more important than counseling them about stress. Seventy-six percent lacked confidence in their ability to counsel patients about stress. The majority of respondents (57%) "rarely" or "never" practiced stress reduction techniques themselves. Belief in the importance of stress counseling, its effectiveness in improving health, and confidence in one's ability to teach relaxation techniques were all related to the probability that providers would counsel patients regarding stress. There is a need for curriculum reform that emphasizes new knowledge about stress and disease, new skills in stress reduction, and more positive beliefs about mind/body medicine and its integration into the existing health care structure. PMID- 14527052 TI - Overlooked role of African-American males' hypermasculinity in the epidemic of unintended pregnancies and HIV/AIDS cases with young African-American women. AB - This article looks at multiple lines of converging evidence relevant to the 72% unintended pregnancy rate, and recently emerged heterosexually-based HIV/AIDS epidemic with young African-American women. Evidence recently reveals a convergence of these epidemics, in a vulnerable subpopulation segment of African American women. Overlooked, as a unique contributing factor in these epidemics is the hypermasculine behaviors of African-American males. Among the risky behaviors linked with this hypermasculinity are a greater tendency with African-American males to have more multiple sexual partners, and a stronger aversion to condom use than other male ethnic groups. As a contributing factor in these epidemics, African-American males' hypermasculinity has several implications for intervention strategies to reduce the epidemics, which are discussed. PMID- 14527053 TI - Utilization of health care resources by HIV-infected white, African-American, and Hispanic men in the era before highly active antiretroviral therapy. AB - In the highly active antiretroviral therapy (HAART) era, U.S. African-American and Hispanic patients with HIV use HAART less, but emergency and inpatient services more, than white patients. We evaluated whether these patterns existed in the pre-HAART era. Data from prospective cohort studies of 462 male Veterans Affairs patients and 1,309 male patients from the AIDS Costs and Services Utilization Survey were combined. Resource utilization of white, African American, and Hispanic men was compared. Compared to whites, African Americans were more likely to visit the emergency department and less likely to have mental health, home health, and dental visits; had fewer outpatient and substance abuse treatment visits; and had more inpatient nights. Hispanics were less likely to have mental health and home health visits, and had more inpatient nights. Whites used prescription drugs more than African Americans or Hispanics, but antiretrovirals were equally used. Lower access to HAART for African-American and Hispanic patients is a new phenomenon, not a continuation of pre-HAART patterns, while the undesirable patterns of emergency and outpatient provider resource utilization in the HAART era are a continuation of pre-HAART patterns. Undesirable resource utilization patterns by African-American and Hispanic populations need urgent attention. PMID- 14527054 TI - Right-sided chest-lead abnormalities on EKG in acute pulmonary embolism. PMID- 14527055 TI - Beginning of AIDS. PMID- 14527057 TI - Cardiac diphtheria in a previously immunized individual. AB - A previously healthy 19-year-old Asian female without significant past medical history presented to the emergency room complaining of a sore throat, difficulty in swallowing, fever, swollen neck, malaise, and myalgia for three to four days. The patient was initially seen at an outside hospital, evaluated by an ear, nose, and throat physician (ENT), and was found to have desquamative pharyngitis. The patient was transferred to our hospital after she continued to experience progressively worsening shortness of breath and went into acute respiratory distress. The patient was found to have laryngeal edema on exam with greenish black, necrotic-looking tissue extending to the hypopharynx, nasopharynx, and oropharynx. A culture was taken. ENT was consulted for tracheostomy placement. The patient refused to have tracheostomy placed. She went into severe respiratory distress and required urgent tracheostomy. A cardiac consult was obtained. A 2D echocardiogram performed one day after admission revealed an ejection fraction (EF) of 10-20%, normal left ventricular cavity size, normal wall thickness, and severe global systolic dysfunction. There was mild to moderate mitral regurgitation and trace tricuspid regurgitation. The inferior vena cava was dilated and a 1 cm x 1.5 cm questionable mass or thrombus was seen. The patient's throat culture was positive for diphtheria. The CDC was contacted, and the patient was treated with antitoxin with prompt resolution of cardiac symptoms. A repeat echo done five days post-treatment showed improved EF of 65%, normal left ventricular thickness and function, with no clot visualized. She was treated with ceftriaxone and flagyl for ocular motor neuritis, otitis media, and strep. pneumonia with gradual improvement. These were all secondary to the diphtheria toxins, however, the patient continues to be followed as an outpatient by ENT for ongoing problems with swallowing, speech, and trach management. PMID- 14527056 TI - HCV in sickle cell disease. AB - The sickle cell gene is common in the U.S. In fact 8% of African Americans are healthy carriers of the sickle cell trait (HbAS). People who are homozygous (HbSS) have severe disease. They have life-long anemia, chronic hemolysis, and also have at times hematological crises, which can worsen the anemia. Many patients require chronic transfusions and as a result, substantial proportions of sickle cell patients are at high risk for infection with blood-borne diseases such as Hepatitis C Virus infection (HCV). The HCV antibody positivity is directly related to the number of transfusions given, and on average the prevalence rate in transfused patients is more than 10%. It is known that the combination of iron overload and HCV can lead to a more rapidly progressive liver disease. The treatment of HCV in sickle cell patients poses a challenge to clinicians. A novel approach described by some is the pre-treatment of these patients with hydroxyurea to increase the fetal hemoglobin, therefore decreasing the severity of Ribavirin-related hemolysis. Treatment with Peg-interferon alone has not been used to treat HCV in sickle cell patients, but in the setting of controlled clinical trials it would be feasible. This review explores the impact of HCV in sickle cell patients and the possible therapeutic options available to them. PMID- 14527058 TI - National Museum of Dentistry exhibition: the future is now! African Americans in dentistry. AB - Inspired by recently published NDA II: The Story of America's Second National Dental Association and sponsored jointly by the National Dental Association Foundation and the Colgate-Palmolive Company, an historical exhibit on dentistry in the African-American community was one of the celebrations for the Golden Anniversary of the American Academy of the History of Dentistry. This exhibit premiered on Sept. 27, 2002 in the National Museum of Dentistry located on the medical/dental campus of the University of Maryland in Baltimore. The Museum recently became an affiliate of the Smithsonian Institution. Contents of the exhibit were photographs, charts, artifacts, memorabilia, etc. These materials presented an overview of African-American activities in dental education, research, patient care, general practice, dental specialities, military service, and public health. Also included were inter-racial relationships, socioeconomic developments, and participation in civil rights endeavors that played a major role in changing out-dated accepted customs. The exhibit's purpose was to celebrate dentistry's ministrations as a health professional among African Americans in particular and the nation at large over the past two centuries. Respect for and progress of black dentists paralleled that of black physicians who were instrumental in including dentist and pharmacists as equal members in the National Medical Association since the latter's inception in 1895. PMID- 14527059 TI - State-of-the-art imaging of acute pancreatitis. AB - The purpose of this essay is to focus on the characteristic imaging features, as revealed by a variety of radiological modalities, which allow for an accurate diagnosis and staging of acute pancreatitis. In addition, emphasis will be made on the role of imaging in providing early prognostic information about the outcome of the disease. PMID- 14527060 TI - Struma ovarii: US and CT findings. AB - We describe the ultrasound and CT findings in a case of struma ovarii. Ultrasound showed a multicystic mass with a well-vascularized solid part. CT demonstrated a multilocular cystic mass with calcifications and solid, enhancing components. To prevent radical surgery, struma ovarii should be included in the differential diagnosis when a mixed, multilocular, teratoma-like tumour of the ovary shows a well-vascularized, enhancing, central solid component. PMID- 14527061 TI - Colour duplex sonographic and multislice spiral CT angiographic diagnosis of ulnar artery aneurysm in hypothenar hammer syndrome. AB - We report a case of hypothenar hammer syndrome secondary to a thrombosed aneurysm of the ulnar artery in a 36-year-old patient presenting with pain due to compression of the ulnar nerve in the Guyon's canal. The initial diagnosis was made by colour Duplex sonography but complete assessment of the thrombosed aneurysm was performed by multislice spiral CT angiography (msCTa). This case illustrates the potential of msCT as an alternative to conventional arteriography. PMID- 14527062 TI - Volume rendering in clinical practice. a pictorial review. AB - Several post-processing techniques are currently available to the radiologist to optimize the review of the scan data acquired by multidetector CT. This is sometimes necessary as, when reviewing a volumetric data acquisition only as transaxial CT images, all the gathered information is not always displayed. Among all the current available post-processing possibilities, volume rendering is one of the most powerful techniques due to its various parameters and powerful segmentation capabilities. It is nevertheless the most complex technique, requiring a higher degree of training and experience from the radiologist to generate the desired result. Aim of this paper is to present the reader a pictorial review of the usefulness of volume rendering as a clinical tool, with emphasis on CT angiography and skeletal pathology. PMID- 14527063 TI - Sequences and techniques in spinal MR imaging. AB - The optimal protocol in spinal MR imaging is not evident. Sagittal T2 weighted FSE, sagittal T1 weighted SE and axial T2 weighted FSE sequences are widely accepted for imaging patients with sciatica and/or lumbar pain. Because of the limited amount of CSF compared to the lumbar spine, the choice of sequences is much more complex in the study of the cervical spine. Sagittal T2 FSE, sagittal T1 SE and axial 2D GE images are suggested in routine cervical spine imaging. To assess the bone marrow, a STIR sequence can be added to this protocol on both lumbar and cervical spine examinations. The 2D GE produces an acceptable image quality to differentiate between the disc and bony protrusions. The use of FLAIR for imaging spinal cord lesions remains controversial in the literature. PMID- 14527064 TI - Pattern recognition of degenerative disorders in the lumbar spine. Guidelines to MR image interpretation. AB - Interpreting MR images of the lumbar spine remains a formidable diagnostic challenge, especially in patients with degenerative changes. What are the pertinent findings? Which abnormalities are clinically relevant? Which changes should be viewed as normal aging? The purpose of this presentation is to present a standardized 7-step approach towards interpreting MR images of the degenerative lumbar spine. PMID- 14527065 TI - Lumbosacral interspinous ligament rupture associated with acute intrinsic spinal muscle degeneration. AB - OBJECTIVE: To demonstrate lumbosacral interspinous ligament rupture, with or without related paraspinal muscle degeneration. SUBJECTS AND METHODS: This study consisted of a prospective imaging analysis of 100 consecutive MRI studies in adult patients (mean age: 56 years) presenting with low back pain. Alterations from the normal in the inter- and perispinal structures of the spine and perispinal soft tissues (e.g. spinal ligaments, perispinal muscles) were sought based upon studies on young volunteers without low back pain (n = 10; mean age: 23 years). RESULTS: Compared to the group without low back pain, many index cases (n = 71: 71%) demonstrated hyperintensity (i.e., sprain or frank ligamentous rupture) of the interspinous ligament(s) on T2-weighted, fat suppressed MRI studies at one (n = 20/71:28%) or multiple (n = 51/71:72%) levels. Associated paraspinal muscle (e.g., interspinalis, multifidus muscles) degeneration was observed in a minority of cases overall (n = 7: 7%), but was only seen in association with cases also demonstrating interspinous ligament degeneration/rupture (n = 7/71:10%). CONCLUSION: Lumbosacral interspinous ligament sprain or frank rupture, as well as related acute-subacute autotraumatic paraspinal muscle rupture/degeneration, may be overlooked by many observers if fat suppressed, T2-weighted MRI is not acquired. These musculoligamentous alterations are on occasion the only abnormalities recognized on MRI of the lumbosacral spine and may theoretically be sources of low back morbidity that potentially may respond to specific therapy. Because this study was an observational one, based solely upon medical imaging, future research must focus upon the correlation of the relevance of these findings with an age-matched asymptomatic control group and longitudinal clinicoradiologic therapeutic trials. PMID- 14527066 TI - Cervical spine fractures and soft tissue injuries. AB - Spinal cord injury is the major cause of quadriplegia and disability. Plain radiographs have a low sensitivity for identifying traumatic cervical spine lesions. Therefore trauma victims with plain films negative for cervical injury but with a high clinical suspicion of injury, or positive for cervical injury should undergo CT or MR for a more definitive evaluation of the cervical spine. Besides the higher sensitivity than plain radiography in detecting fractures, CT is also able to show soft-tissue abnormalities. MR is the definitive modality in assessing cervical soft-tissue injuries, especially in the evaluation of the spinal cord, intervertebral discs, and ligaments. It also allows differentiate spinal cord hemorrhage and edema, which may have a prognostic value. The role of medical imaging in the evaluation of whiplash injuries remains to be determined. PMID- 14527068 TI - Interrupted right pulmonary artery. PMID- 14527067 TI - Ultrasound and computed tomography: spin-offs of the world wars. AB - Important losses to the ships of the allied troops by the attacks of the German submarines during World War I led researchers to find specific detecting devices as a means of defence. In 1880 Pierre Curie and his brother, discovered the production of ultrasound waves. Langevin, their student, applied this invention to the localisation of boats. At the end of WWI, research and results ended up being forgotten, but gained attention again with the sonar when WWII loomed on the horizon. At the end of the war, a former military medical doctor, G. Ludwig (US Navy), tried to localize gallstones with a left-over sonar apparatus. This definitely led to firm conclusions. Other researchers in several countries contributed to refining this new imaging technique which is nowadays widely applied. During WWII, the American and British army developed considerable research in the field of the calculator (computer) to speed up deciphering the secret codes. Coupling the principles of tomography discovered during WWI with the computing capability of the calculators developed during WWII, computerized axial tomography could be obtained. This new technology, which is used daily, probably is one of the greatest acquisitions of the 20th century in the field of medical imaging. PMID- 14527069 TI - Spontaneous splenic capsule rupture complicating infectious mononucleosis. PMID- 14527070 TI - Imaging of congenital abnormalities of the skeleton. PMID- 14527071 TI - How I remember Bill. PMID- 14527072 TI - Principles from evolutionary biology and psychology will further strengthen tenets and profession. PMID- 14527073 TI - Reflections of a third-year medical student. PMID- 14527074 TI - Are we allowing the mature development of competency in medical students and residents? PMID- 14527075 TI - Questions raised regarding fund reporting. PMID- 14527076 TI - Osteopathic manipulative treatment in the emergency department for patients with acute ankle injuries. AB - STUDY OBJECTIVE: The purpose of this study was to evaluate the efficacy of osteopathic manipulative treatment (OMT) as administered in the emergency department (ED) for the treatment of patients with acute ankle injuries. METHODS: Patients aged 18 years and older with unilateral ankle sprains were randomly assigned either to an OMT study group or a control group. Independent outcome variables included edema, range of motion (ROM), and pain. Both groups received the current standard of care for ankle sprains and were instructed to return for a follow-up examination. Patients in the OMT study group also received one session of OMT from an osteopathic physician. RESULTS: Patients in the OMT study group had a statistically significant (F = 5.92, P = .02) improvement in edema and pain and a trend toward increased ROM immediately following intervention with OMT. Although at follow-up both study groups demonstrated significant improvement, patients in the OMT study group had a statistically significant improvement in ROM when compared with patients in the control group. CONCLUSIONS: Data clearly demonstrate that a single session of OMT in the ED can have a significant effect in the management of acute ankle injuries. PMID- 14527077 TI - Effect of topical nasal corticosteroids on patients with chronic fatigue syndrome and rhinitis. AB - BACKGROUND: Chronic fatigue syndrome (CFS) is a disabling illness of persistent fatigue. Recent studies have shown that patients with CFS have an increased prevalence of nonallergic rhinitis. Inflammation of the nasal passages due to allergic rhinitis can cause nasal congestion resulting in an increased number of sleep disturbances and daytime fatigue. While topical nasal corticosteroids have been shown to alleviate nasal obstruction effectively in patients with rhinitis who do not have CFS, it is unknown whether topical nasal corticosteroids will reduce CFS symptoms. STUDY OBJECTIVE: The purpose of this study is to determine whether topical nasal corticosteroids will reduce daytime sleepiness in patients with CFS and rhinitis. METHODS: Twenty-eight of 31 subjects with rhinitis and a diagnosis of CFS completed the double-blind, randomized, placebo-controlled trial. Two subjects failed screening, and 3 subjects withdrew from the study prior to its completion. Subjects were randomized according to Balaam's crossover design, and one of the following interventions was used for each group in the study: 8-week treatment with a topical nasal corticosteroid, 8-week treatment with a placebo saline spray, 4-week treatment with a topical nasal corticosteroid followed by a 4-week treatment with a placebo saline spray, or a 4-week treatment with a placebo saline spray followed by a 4-week treatment with a topical nasal corticosteroid. Data focusing on rhinitis symptoms, severity of chronic fatigue symptoms, and quality of life were gathered at biweekly office visits and with daily diaries. RESULTS: The results indicated that daytime sleepiness was reduced when patients with rhinitis and CFS were treated with topical nasal corticosteroids. The severity of associated CFS symptoms, specifically fatigue, muscle pain, postexertional fatigue, and daily activity, did not improve with treatment. CONCLUSION: Treating the symptoms of rhinitis in patients with CFS does not appear to alleviate daytime fatigue or associated nasal, musculoskeletal, or cognitive complaints. Therefore, it is unlikely that aggressive treatment of such symptoms with topical nasal corticosteroids will provide significant benefit to patients with CFS who do not have allergic rhinitis. These results indicate that the nonallergic rhinitis seen in patients with CFS may arise from a mechanism other than chronic inflammation. PMID- 14527078 TI - Development of the Attitudes Toward Osteopathic Principles and Practice Scale (ATOPPS): preliminary results. AB - Little empirical work has been done to examine how osteopathic medical students' attitudes toward osteopathic principles and practice (OPP) develop and evolve over the course of their medical education. A major obstacle to conducting this research is the absence of reliable and sensitive instruments to measure students' attitudes toward OPP. The purpose of this project is to develop a sensitive and reliable instrument to measure students' attitudes toward OPP. Face to-face and telephone interviews were conducted with osteopathic medical students, osteopathic manipulative medicine (OMM) residents, OMM undergraduate fellows, and three board-certified OMM specialists. These interviews were summarized in a 39-item instrument administered to 127 students at the completion of their core OMM rotation at the University of North Texas Health Science Center at Fort Worth-Texas College of Osteopathic Medicine. Factor analysis of student responses to the 39 candidate items yielded two interpretable factors. Factor 1 contained 24 items and accounted for 33% of the item response variance, and factor 2 contained four items and accounted for 5.6% of the item response variance. Based on these results, 14 of the original 39 statements were eliminated and the smaller second factor was dropped. Factor 1 contained items reflecting both positive and negative attitudes toward the application of OPP in patient care, the importance of OPP in medical education, and professional distinctiveness. One of the original 39 candidate items was returned to factor 1 because it was judged by the investigators to be consistent with the underlying construct of the scale and helped balance the number of forward-scored and reversed-scored items in the final instrument. Two internal consistency estimates of reliability were computed for the revised 25-item Attitudes Toward Osteopathic Principles and Practice Scale (ATOPPS): the Spearman-Brown unequal-length corrected coefficient alpha and the split-half reliability coefficients. Estimates for the split-half coefficients were .89 for part 1 and .87 for part 2. The Spearman-Brown coefficient alpha was .93, indicating substantial internal consistency. The 25-item ATOPPS seems to reflect a continuum of positive and negative attitudes toward OPP. This preliminary report documents reliability for the 25-item ATOPPS. With continued support for its construct validity, ATOPPS provides investigators with a reliable tool to assess the development of attitudes toward OPP. PMID- 14527079 TI - Productivity outcomes for recent grants and fellowships awarded by the American Osteopathic Association Bureau of Research. AB - The objective of the present study was to evaluate productivity outcome measures for recent research grants and fellowships awarded through the American Osteopathic Association (AOA) Bureau of Research. Recipients of grants and fellowships that were awarded between 1995 and 2001 were contacted by mail, e mail, or telephone and asked to provide information about publications, resulting grant awards, advances in clinical care, or other notable products that were generated from their projects. For grants funded between 1995 and 1998, 76% of principal investigators reported a notable product from their study. By contrast, for grants funded between 1999 and 2001, only 31% reported a notable outcome. This difference most likely can be attributed to the lag time between the awarding of a grant and actual completion of the project, the processing of the data, and the publication of the results. Several recipients of 1999-2001 grants were optimistic about eventually generating a notable product. Most (79%) of the 1995-2001 fellows met the requirements for successful completion of their project. Many fellows exceeded the minimal requirement by publishing their results, continuing research activity, attracting extramural grant monies, or entering an academic position. It appeared that a much larger proportion of osteopathic fellows went on to academic careers than their counterparts who did not have fellowship training. From 1995 to 2001, the AOA Bureau of Research awarded dollars 3,072,140 in research grants and fellowships. To date, these awards have helped the recipients bring an additional dollars 5,659,329 of extramural funds for research at osteopathic institutions. The Bureau of Research grant and fellowship programs have been successful both scientifically and in terms of financial outcomes. PMID- 14527080 TI - Mechanisms of liver injury: an overview. AB - Liver cirrhosis, an end-result of a wide variety of the liver diseases, is a world wide health problem. Because of its unique organ system, i.e., portal blood supply, bile formation and enterohepatic circulation, drug metabolism system, and sinusoidal lining cells such as Kupffer, endothelial and stellate cells, the liver is a target of a variety of hepatotoxic insults. Current data suggest that hepatocyte apoptosis is an essential feature contributing to liver injury in a wide range of acute and chronic liver diseases. With an improved understanding of the pathophysiological role of apoptosis in liver diseases, we are now entering an era where regulation of liver cell apoptosis is becoming a therapeutic possibility. Inhibition of hepatocyte apoptosis using a variety of different strategies may be therapeutically beneficial in liver injuries, such as alcoholic hepatitis, non-alcoholic steatohepatitis (NASH), viral hepatitis, and cholestatic liver diseases. Considering the link between hepatocyte apoptosis and liver fibrosis, inhibition of hepatocyte apoptosis may also be an anti-fibrotic therapeutic strategy. Moreover, selective induction of apoptosis of activated stellate cells would be a unique approach to induce the resolution the phase of liver fibrosis. These concepts merit further clinical and basic investigation. PMID- 14527081 TI - Death receptor activation-induced hepatocyte apoptosis and liver injury. AB - The TNFalpha receptor super-family consists of several members sharing a sequence homology in a unique function domain, the death domain, which is located in the intracellular portion of the receptor. These so-called death receptors, including Fas, TNF-R1 and TRAIL-R1/TRAIL-R2, are expressed on hepatocytes. When stimulated by their ligands, FasL, TNFalpha or TRAIL, respectively, the death receptors can activate multiple death domain-initiated apoptosis programs, including both extrinsic and intrinsic pathways. A cascade of caspases is activated, which cleave proteins important for the cell structure and function. Activation of the intrinsic pathway also leads to mitochondrial release of several apoptotic proteins and mitochondrial dysfunction, which kill the cell through both caspase dependent and caspase-independent mechanisms. Death receptor-induced hepatocyte apoptosis contributes to the development of a number of liver diseases, including viral hepatitis, inflammatory hepatitis, Wilson's disease, alcoholic liver disease, endotoxiemia-induced liver failure and ischemia/reperfusion-induced liver damage. This article comprehensively reviews the mechanisms of induction and regulation of death receptor-initiated apoptosis in hepatocytes, examines how these molecular events affect our understanding of the pathogenesis of these diseases and further discusses the potential therapeutic application of the knowledge. We hope we can provide a cohesive and integrated perspective on the many aspects of these complicated processes. PMID- 14527082 TI - CYP2E1: biochemistry, toxicology, regulation and function in ethanol-induced liver injury. AB - Ethanol-induced oxidative stress appears to play a major role in mechanisms by which ethanol causes liver injury. Many pathways have been suggested to contribute to the ability of ethanol to induce a state of oxidative stress. One central pathway appears to be the induction of the CYP2E1 form of cytochrome P450 enzymes by ethanol. CYP2E1 is of interest because of its ability to metabolize and activate many toxicological substrates, including ethanol, to more reactive, toxic products. Levels of CYP2E1 are elevated under a variety of physiological and pathophysiological conditions, and after acute and chronic alcohol treatment. CYP2E1 is also an effective generator of reactive oxygen species such as the superoxide anion radical and hydrogen peroxide, and in the presence of iron catalysts, produces powerful oxidants such as the hydroxyl radical. This Review Article summarizes some of the biochemical and toxicological properties of CYP2E1, and briefly describes the use of HepG2 cell lines developed to constitutively express the human CYP2E1 in assessing the actions of CYP2E1. Regulation of CYP2E1 is quite complex and will be briefly reviewed. Possible therapeutic implications for treatment of alcoholic liver injury by inhibition of CYP2E1 or CYP2E1-dependent oxidative stress will be discussed, followed by some future directions which may help to understand the actions of CYP2E1 and its role in alcoholic liver injury. PMID- 14527083 TI - Role of nitric oxide in liver injury. AB - The complex role of nitric oxide (NO) in the liver can be explained by its patterns of regulation and unique biochemical properties. With a broad range of direct and indirect molecular targets, NO acts as an inhibitor or agonist of cell signaling events. In the liver, constitutively generated NO maintains the hepatic microcirculation and endothelial integrity, while inducible NO synthase (iNOS) governed NO production can be either beneficial or detrimental. For instance, NO potentiates the hepatic oxidative injury in warm ischemia/reperfusion, while iNOS expression protects against hepatic apoptotic cell death seen in models of sepsis and hepatitis. Anti-apoptotic actions are either cyclic nucleotide dependent or independent, including the expression of heat shock proteins, prevention of mitochondrial dysfunction, and inhibition of caspase activity by S-nitrosation. Whether NO protects or injures is probably determined by the type of insult, the abundance of reactive oxygen species (ROS), the source and amount of NO production and the cellular redox status of liver. Through the use of pharmacological NO donors or NOS gene transfer in conjunction with genetically altered knockout animals, the physiological and pathophysiological roles of NO in liver function can be explored in more detail. The purpose of this paper is to review the current understanding of the role of NO in liver injury. PMID- 14527084 TI - Role of the mitochondrial permeability transition in apoptotic and necrotic death after ischemia/reperfusion injury to hepatocytes. AB - Reperfusion of ATP-depleted tissues after warm or cold ischemia causes pH dependent necrotic and apoptotic cell death. In hepatocytes and other cell types as well, the mechanism underlying this reperfusion-induced cell death involves onset of the mitochondrial permeability transition (MPT). Opening of permeability transition (PT) pores in the mitochondrial inner membrane initiates the MPT, an event blocked by cyclosporin A (CsA) and pH less than 7.4. Thus, both acidotic pH and CsA prevent MPT-dependent reperfusion injury. Glycine also blocks reperfusion induced necrosis but acts downstream of PT pore opening by stabilizing the plasma membrane. After the MPT, ATP availability from glycolysis or other source determines whether cell injury after reperfusion progresses to ATP depletion dependent necrosis or ATP-requiring apoptosis. Thus, apoptosis and necrosis after reperfusion share a common pathway, the MPT. Cell injury progressing to either necrosis or apoptosis by shared pathways can be more aptly termed necrapoptosis. PMID- 14527085 TI - Mechanisms of viral hepatitis induced liver injury. AB - Among seven human hepatitis viruses (A to E, G and TT virus), hepatitis B (HBV) and C (HCV) viruses are able to persist in the host for years and principally contribute to the establishment of chronic hepatitis. During the course of persistent infection, continuous intrahepatic inflammation maintains a cycle of liver cell destruction and regeneration that often terminates in hepatocellular carcinoma (HCC). While the expression and retention of viral proteins in hepatocytes may influence the severity and progression of liver disease, the mechanisms of liver injury in viral hepatistis are defined to be due not to the direct cytopathic effects of viruses, but to the host immune response to viral proteins expressed by infected hepatocytes. In the process of liver injury, hepatocellular death (apoptosis) induced by the proapoptotic molecules of T cells activated following antigen recognition triggers a cascade of antigen nonspecific effector systems and causes necroinflammatory disease. Accordingly, the regulation of the immune response, e.g., via the cell death pathways, in chronically infected patients should prevent the development of HCC. PMID- 14527086 TI - Cytokine-induced inflammatory liver injuries. AB - IL-18 is a pleiotropic cytokine and is produced by various types of cells including activated macrophages, particularly Kupffer cells. IL-18 has potential to activate inflammatory responses through induction of IFN-gamma production in collaboration with IL-12. Somewhat paradoxically, IL-18 also has the capacity to induce allergic responses via induction of IL-4 production by T helper cells and to activate mast cells and basophils to release atopic effector molecules such as histamine. Indeed, IL-18 is involved in inflammatory tissue injuries, such as Crohn's disease and atherosclerosis, and also in hyper IgE and atopic dermatitis. IL-18 is particularly important for induction of experimental liver diseases. Endotoxin-induced liver injury or Fas ligand-induced hepatitis is caused by endogenous IL-18 in mice. Moreover, patients with liver diseases such as fulminant hepatitis, liver cirrhosis due to hepatitis virus infection and primary biliary cirrhosis show elevation of serum levels of IL-18, that correlates with the corresponding disease severity. Therefore, endogenous IL-18 plays a major role in induction of some types of liver injuries in mice and human. NKT cells that express both T cell receptor and NK cell marker are abundant in the liver of mice and human. Recent studies have revealed that NKT cells participate in some types of liver injuries, such as concanavalin A-induced T cell-mediated hepatitis and malaria hepatitis. In this review article, we focus on IL-18-involving liver damages and NKT-cell-mediated liver injuries. PMID- 14527087 TI - Peroxisome proliferator-activated receptors, fatty acid oxidation, steatohepatitis and hepatocarcinogenesis. AB - Fatty acids are metabolized in the liver by beta-oxidation in mitochondria and peroxisomes and by omega-oxidation in microsomes. Peroxisomal beta-oxidation is responsible for the metabolism of very long chain fatty acids and mitochondrial beta-oxidation is responsible for the oxidation of short, medium and long chain fatty acids. Very long chain fatty acids are also metabolized by the cytochrome P450 CYP4A omega-oxidation system to dicarboxylic acids. Both peroxisomal beta oxidation and microsomal omega-oxidation lead to the generation of H2O2. The genes encoding peroxisomal, microsomal and some mitochondrial fatty acid metabolizing enzymes in the liver are transcriptionally regulated by peroxisome proliferator-activated receptor alpha (PPAR alpha). Sustained activation of PPAR alpha by peroxisome proliferators has been shown to induce hepatocellular carcinomas in rats and mice. The peroxisome proliferator-induced carcinogenic effect has been attributed to transcriptional activation of PPAR alpha regulated genes and the resulting excessive generation of H2O2. Evidence from mice lacking fatty acyl-CoA oxidase (AOX), PPAR alpha and PPAR alpha/AOX has confirmed the role of PPAR alpha in the development of hepatocellular carcinomas. In addition, mice lacking AOX developed steatohepatitis and provided clues regarding the molecular mechanism responsible for steatosis and steatohepatitis and the role of unmetabolized AOX substrates in the activation of PPAR alpha. PMID- 14527088 TI - Mechanisms of human hepatocarcinogenesis. AB - The major risk factors and etiological agents responsible for development of hepatocellular carcinoma in humans have been identified and characterized. Among these are chronic infection with hepatitis B virus or hepatitis C virus, exposure to aflatoxin B1, and cirrhosis of any etiology (including alcoholic cirrhosis and cirrhosis associated with genetic liver diseases). Both chronic hepatitis and cirrhosis represent major preneoplastic conditions of the liver as the majority of hepatocellular carcinomas arise in these pathological settings. Hepatocarcinogenesis represents a linear and progressive process in which successively more aberrant monoclonal populations of hepatocytes evolve. Regenerative hepatocytes in focal lesions in the inflamed liver (chronic hepatitis or cirrhosis) give rise to hyperplastic hepatocyte nodules, and these progress to dysplastic nodules, which are thought to be the direct precursor of hepatocellular carcinoma. In most cases, the neoplastic transformation of hepatocytes results from accumulation of genetic damage during the repetitive cellular proliferation that occurs in the injured liver in response to paracrine growth factor and cytokine stimulation. Hepatocellular carcinomas exhibit numerous genetic abnormalities (including chromosomal deletions, rearrangements, aneuploidy, gene amplifications, and mutations), as well as epigenetic alterations (including modulation of DNA methylation). These genetic and epigenetic alterations combine to activate positive mediators of cellular proliferation (including cellular proto-oncogenes and their mitogenic signaling pathways) and inactivate negative mediators of cellular proliferation (including tumor suppressor genes), resulting in cells with autonomous growth potential. However, hepatocellular carcinomas exhibit a high degree of genetic heterogeneity, suggesting that multiple molecular pathways may be involved in the genesis of subsets of hepatocellular neoplasms. Continued investigation of the mechanisms of hepatocarcinogenesis will refine our current understanding of the molecular and cellular basis for neoplastic transformation in liver, enabling the development of effective strategies for prevention and/or more effective treatment of hepatocellular carcinoma. PMID- 14527089 TI - Disposition of quercetin and kaempferol in human following an oral administration of Ginkgo biloba extract tablets. AB - Ten adult volunteers with an average age 28 years were given a single oral dose of six tablets of Ginkgo biloba extract. Quercetin and kaempferol in different period of human urine were determined by using RP-HPLC. The results showed the elimination rate constant k and the absorption rate constant ka of quercetin were slightly more than that of kaempferol; and the absorption half-life (t(1/2a)), the elimination half-life (t(1/2)) and t(max) of quercetin were less than that of kaempferol, the differences were, however, not statistically significant. The mean values of ka were 0.61 h(-1) and 0.55 h(-1), t(1/2a) 1.51 h and 1.56 h, k 0.37 h(-1) and 0.30 h(-1), t(1/2) 2.17 h and 2.76 h, T(max) 2.30 h and 2.68 h for quercetin and kaempferol, respectively, which mean absorption and elimination of quercetin and kaempferol are 0.17% and 0.22%, respectively. Quercetin and kaempferol are excreted in the human urine mainly as glucuronides. PMID- 14527090 TI - Effect of Tamarindus indica. L on the bioavailability of ibuprofen in healthy human volunteers. AB - The influence of Tamarindus indica L fruit extract incorporated in a traditional meal on the bioavailability of Ibuprofen tablets 400 mg dose when given concurrently was studied in 6 healthy human volunteers. There was a statistically significant increase in the plasma levels of Ibuprofen and its metabolites hydroxy-ibuprofen and carboxy-ibuprofen respectively, when the meal containing Tamarindus indica fruit extract was administered with the ibuprofen tablets than when taken under fasting state or with the meal without the fruit extract. The C(max), AUC(0-6 hr) and Ka for ibuprofen increased from 38 +/- 0.70 microg/ml to 42 +/- 0.98 microg/ml (p > 0.05); and 28.03 +/- 2.40 microg/ml x hr to 56.51 +/- 0.16 microg/ml x hr (p < 0.05) and 1.048 +/- 0.02hr(-1) to 2.781 +/- 0.11 hr(-1) (p < 0.05) respectively. There was no change in the t(max) (120.00 +/- 0.43m) but there was a decrease in the k(el) from 0.63 +/- 0.20 hr(-1) to 0.46 +/- 0.11 hr( 1) (p<0.05). Similarly the C(max), AUC(0-6 h) and Ka for hydroxy-ibuprofen rose from 43 +/- 0.76 microg/ml to 45 +/- 0.16 microg/ml (p < 0.05); 39.04 +/- 2.30 microg/ml x hr to 59.49 +/- 2.39 microg/ml.hr in (p < 0.05) and 1.498 +/- 0.79hr( 1) to 3.442 +/- 0.23 hr(-1) (p < 0.05) respectively; while the C(max), AUC(0-6 h) and Ka for carboxy-ibuprofen rose from 48 +/- 0.7 microg/ml to 51 +/- 0.16 microg/ml (p < 0.05); 41.972 +/- 0.68 microg/ml x hr to 63.948 +/- 0.12 microg/ml x hr (p < 0.05) and 1.649 +/- 0.08 hr(-1) to 4.187 +/- 0.42 hr(-1) (p < 0.05) respectively. The study has indicated that Tamarindus indica L. fruit extract significantly increased the bioavailability of Ibuprofen. PMID- 14527091 TI - The effect of four different types of food on the bioavailability of cefaclor. AB - This randomized, open-label, balanced, five-treatment, five-period, five sequence, single-dose and crossover pharmacokinetic study assessed the effect of different types of food on the bioavailability of cefaclor in 18 healthy male volunteers. A single dose of cefaclor, 250-mg capsule was administered at five occasions: after overnight fasting, after two vegetarian (high-fat and low-fat) diets and two non-vegetarian (high-fat and low-fat) diets. Serial blood samples were collected upto 8 h post dose. Serum cefaclor concentrations were determined by a validated HPLC method. AUC values were not significantly affected by food intake, but the T(max) was prolonged and C(max) was decreased, depending on the type of meal. The non-vegetarian diets affected the rate of absorption of cefaclor more than the vegetarian diets. The least decrease in C(max) was produced by low-fat vegetarian diet, while the maximum decrease was produced by high-fat non-vegetarian diet. The results of this study indicate that while the rate of absorption of cefaclor is significantly decreased, the extent of absorption and the rate of elimination are not significantly decreased in the presence of food. As compared to high-fat non-vegetarian diet, the time above MIC50 concentration was significantly increased by low-fat vegetarian diet. The implications of these findings for the large vegetarian Indian population are considerable. PMID- 14527092 TI - Comparison of stepwise and simultaneous estimations of population pharmacokinetics and pharmacodynamics of TS-943. AB - The prediction performances of population pharmacokinetic-pharmacodynamic analysis of the two methods (a stepwise and a simultaneous estimations) were evaluated with respect to their accuracies and precisions. A study was designed to investigate the safety and efficacy of TS-943 by a 4 hours constant infusion in 36 healthy male subjects. Population analysis was performed using pharmacokinetic and pharmacodynamic models with NONMEM. The mean of the prediction error (MPE) and the root mean squared error (RMSE) served as a measure of accuracy and precision. In addition, a bootstrap validation was also performed. The results indicate that those population pharmacokinetic pharmacodynamic parameters for the two methods were comparable. The results of simultaneous estimations are similar to those obtained using a stepwise estimation. The mean parameter estimates obtained with the additional 200 bootstrap replicates of data were within 15% of those obtained with the final model in both methods. The present results demonstrated that the accuracy of pharmacodynamic evaluations using a stepwise end a simultaneous estimations was comparable. PMID- 14527093 TI - Absorption, distribution, metabolism and excretion of the cholecystokinin-1 antagonist dexloxiglumide in the rat. AB - Single oral doses of 14C-dexloxiglumide were rapidly and extensively absorbed in rats, and eliminated more slowly by females than by males. The respective half lives were about 4.9 and 2.1 h. Following single intravenous doses, dexloxiglumide was characterised as a drug having a low clearance (6.01 and about 1.96 ml/min/kg in males and females respectively), a moderate volume of distribution (Vss, 0.98 and about 1.1 L/kg in males and females respectively) and a high systemic availability. It was extensively bound to plasma proteins (97%). Dexloxiglumide is mainly cleared by the liver. Its renal clearance was minor. In only the liver and gastrointestinal tract, were concentrations of 14C generally greater than those in plasma. Peak 14C concentrations generally occurred at 1-2 h in males and at 2-4 h in females. Tissue 14C concentrations then declined by severalfold during 24 h although still present in most tissues at 24 h but only in a few tissues (such as the liver and gastrointestinal tract) at 168 h. Decline of 14C was less rapid in the tissues of females than in those of males. Single intravenous or oral doses were mainly excreted in the faeces (87-92%), mostly during 24 h and more slowly from females than from males. Urines contained less than 11% dose. Mean recoveries during 7 days when 14C was not detectable in the carcass except in one female rat ranged between 93-101%. Biliary excretion of 14C was prominent (84-91% dose during 24 h) in the disposition of 14C which was also subjected to facile enterohepatic circulation (74% dose). Metabolite profiles in plasma and selected tissues differed. In the former, unchanged dexloxiglumide was the major component whereas in the latter, a polar component was dominant. Urine, bile and faeces contained several 14C-components amongst which unchanged dexloxiglumide was the most important (eg. up to 63% dose in bile). LC-MS/MS showed that dexloxiglumide was metabolised mainly by hydroxylation in the N-(3 methoxypropyl)pentyl sidechain and by O-demethylation followed by subsequent oxidation of the resulting alcohol to a carboxylic acid. PMID- 14527094 TI - Reproducibility over time of the urinary diclofenac/4'-OH diclofenac ratio among different CYP2C9 genotypes. AB - Diclofenac has been used for the evaluation of CYP2C9 activity in vitro as well as in vivo with varying results. The present study was aimed at evaluating the reproducibility of the urinary diclofenac/4'-OH diclofenac ratio among different CYP2C9 genotypes in healthy volunteers. The study of CYP2C9 genotypes in the family of a CYP2C9*3/*3 subject is also reported. The urinary diclofenac/4'-OH diclofenac ratio was determined on two occasions within a period of 9-12 months, and was found to be correlated (r = 0.83, p < 0.05). The mean (+/- SD) of diclofenac/4'-OH diclofenac ratio was 1.5 times higher among subjects carrying CYP2C9*3 allele (CYP2C9*1/*3 and CYP2C9*2/*3 genotypes) (0.91 +/- 0.28), compared to CYP2C9*1/*1 subjects (0.60 +/- 0.11). The results show that the urinary diclofenac/4'-OH diclofenac ratio might be used to study CYP2C9 in humans. The data agree with previous studies showing that the CYP2C9*3 allelic variant seems to cause a decreased CYP2C9 hydroxylation capacity. PMID- 14527095 TI - Induction of propranolol metabolism in isolated rats hepatocytes treated by di(2 ethylhexyl) phthalate (DEHP) and mono(2-ethylhexyl) phthalate (MEHP). AB - Blood lines of polyvinyl chloride (PVC) for hemodialysis usually contain di(2 ethylhexyl) phthalate (DEHP) as a plasticizer. Previous studies show that 1 mg/kg of this plasticizer can leach into the blood during one dialysis session. It is rapidly metabolized in the liver. Mono(2-ehtylhexyl) phthalate (MEHP), its main metabolite can be detected as well. After oral administration to rodents, both compounds caused a variety of adverse biological effects such as testicular atrophy, peroxisome proliferation and hepatic peroxisomal enzyme induction. Male wistar rats were treated intraperitoneally by DEHP and MEHP using twice the dose of that involved in human exposure during a dialysis session. Propranolol metabolism by hepatocytes was investigated after fresh isolation from treated and untreated rats by means of reverse phase HPLC. The choice of propranolol as a substrate was made because of its rather quick liver metabolisation. Phenobarbital was chosen in the study as a reference of enzymatic inducer to evaluate the inducing effect of DEHP and MEHP. Propranolol was metabolized by the hepatocytes of both treated and untreated rats. Hepatocytes isolated from rats treated by phenobarbital, MEHP and DEHP were shown to have a higher speed constant of metabolism indicating a rapid metabolism of propranolol. Under these conditions, in fact, propranolol metabolisation was found to be respectively 6, 2.7, 2 times faster than the propranolol metabolisation of untreated rats. The hypothesis that DEHP and MEHP are enzymatic inducers, particularly cytochrome P450 (CYP) inducers of the xenobiotics metabolism on the intact liver after IP administration has become been found to be valid. The results obtained in this study confirm the value of isolated hepatocytes as an in vivo drug metabolism predictive model. PMID- 14527096 TI - Metabolic assessment in liver microsomes by co-activating cytochrome P450s and UDP-glycosyltransferases. AB - A "dual-activity" microsomal system in which both CYPs and UGTs were active was evaluated for studies of metabolic stability and in-vitro metabolite profiling. In this "dual-activity" system, alamethicin, a pore-forming peptide, was used to activate UGTs in human liver microsomes without affecting CYP activity. Interference studies indicated that CYP cofactors had little effect on UGT surrogate activity as measured by glucuronidation of acetaminophen and trifluoperazine. Further, UGT cofactor, UDPGA (< 2 mM), did not inhibit the marker activity of five major CYPs including 1A2, 2C9, 2C19, 2D6 and 3A4, suggesting that both oxidation and glucuronidation can be co-activated in microsomes. In a comparison study, compounds with significant glucuronidation showed distinct stability profiles in the "dual-activity" system, compared to the conventional microsomal incubation in which only CYPs were active. For compounds with minor or no glucuronidation, the metabolic stability remained similar between the "dual-activity" system and the conventional microsomal incubation. The feasibility of this "dual-activity" system utilized for metabolite profiling was also investigated using tramadol as a model drug. It was found that oxidative metabolites of tramadol generated in the "dual-activity" system matched those detected in the conventional microsomal incubation. However, tramadol glucuronide was observed in the "dual-activity" system but not in the conventional micromosal incubation. Results clearly suggest that the "dual-activity" system is a valuable in vitro model for metabolism studies in drug discovery. PMID- 14527097 TI - The design of pharmacokinetic studies to support drug discovery: the selection of the optimum number of animals for a study. AB - Using historic data from development and pre-development compounds, the variability in plasma concentration (including C(max)) and AUC that would be observed when only 1 or 2 animals per time point are used instead of the normal three has been studied. The results indicate that for pharmacokinetic studies carried out during the early discovery phase the number of animals per time point can be reduced to 2 without significantly affecting the quality of the data. Such an approach is not only more efficient, but it conserves valuable compound and reduces by one third the number of animals employed on the study. PMID- 14527099 TI - Modifications of clonidine binding to rabbit liver protein under the influence of non-steroid-anti-inflammatory drugs in vitro. AB - This study was designed to investigate the binding of clonidine to liver protein as well as the possible interactions with non-steroid anti-inflammatory drugs (NSAIDs) during the binding process in the rabbit. The binding of clonidine to slices (S) and homogenized slices (H) was estimated by a radioisotopic method following incubation with a mixture of cold and 3H-labelled clonidine in Ringer solution at 37 degrees C for 360 min. The binding of clonidine was assessed in the absence and presence of the following NSAIDs: flurbiprofen, ketoprofen, ibuprofen and acetylsalicylic acid. The results showed that the percentage of clonidine binding did not differ between intact and homogenized slices. The addition of all NSAIDs but ibuprofen, significantly decreased the protein binding of clonidine both in intact and homogenized liver slices. This finding could be attributed to the different affinity of ibuprofen for liver protein compared to the remaining NSAID's which may arise from a number of chemical properties including its dual Pka values. PMID- 14527098 TI - Modulatory influence of tin-protoporphyrin on gossypol-induced alterations of heme oxygenase activity in male Wistar rats. AB - Gossypol--a male contraceptive is toxic and causes anorexia, reduction in body weight, hypokalemia etc. It prevents liberation of oxygen from oxyhemoglobin and has hemolytic effect on erythrocytes and leads to microcytic hypochromic anemia. SnPP has been shown to either competitively suppress or to significantly ameliorate a variety of naturally occuring or experimentally induced forms of jaundice in animals and man by inhibiting heme degradation. In this paper novel tissue-dependent response to differential dosing regimen of gossypol and gossypol in association with Sn-protoporphyrin (SnPP) is described. Gossypol was found to be a stimulator of heme oxygenase activity in the liver and kidney to varying degrees. This tissue response contrasted with that of the spleen, where gossypol decreased the activity of the enzyme. The increase in enzymatic activity was accompanied by a decline in the total microsomal protein content on gossypol administration. The gossypol mediated an increase of heme oxygenase activity, elevated bilirubin levels leading to hyperbilirubinemia. The stimulatory effect of gossypol was counteracted to a considerable extent when SnPP was simultaneously administered. Hence, we envision the importance of combined rather than single exposures in defining the realms of toxicology of these and other related drugs. We further envisage the existence of important gossypol-heme interactions in the regulation of heme metabolism. PMID- 14527100 TI - The occupation of healthcare management: relating core competencies to growth as a distinct profession. AB - There are many questions regarding the education of the healthcare manager. Some would argue that rather than focus on specific issues of core competencies and licensing, the discussion should be directed toward the successful development of healthcare management as a distinct profession. Two elements in defining a profession, education and legal restraints, are particularly relevant to healthcare management. It would be extremely difficult, if not impossible, for healthcare management to achieve recognition as a distinct profession in the traditional sense. There is no unique body of knowledge on which the profession is based, and no mechanism for credentialing to restrict practice. The organizational framework, however, does exist to better position the MHA-prepared graduate as the preferred candidate to manage healthcare organizations. A professional society needs to be identified as the representative society for the generalist healthcare manager. Core competencies should articulate the integration of academics and practice preparation. To accomplish the latter, academic accreditation requirements relating to faculty credentials should encourage practice-based credentials in addition to the current exclusive focus on academic credentials. PMID- 14527101 TI - Revisiting Bloom's taxonomy for ethics and other educational domains. AB - In the process of developing competency-based health services administration education, the Ethics Faculty Forum Co-Chairs from the Association of University Programs in Health Administration (AUPHA) were asked not only to identify their domains and competencies, but also to review six other faculty fora research outcomes. This article was written by the Ethics Faculty Forum Co-Chairs in response to the AUPHA request. Reviewing the work of the original six faculty fora using Bloom's taxonomy, we found that the fora focused mainly on the cognitive objectives and generally did not consider the affective objectives. The intent of this paper is to help those who teach healthcare ethics refine their current courses to include both cognitive and affective objectives. The paper pursues five objectives: 1. review of Bloom's taxonomy as a framework for creating course objectives in both the cognitive and affective domains; 2. present fora research and their domain outcomes; 3. present an overview of healthcare ethics literature; 4. provide a demonstration of healthcare ethics competencies in both the cognitive and affective domains; and 5. present possible directions for healthcare ethics and other educational domain research. PMID- 14527102 TI - Rx for the staffing crisis: bold partnerships not band-aids. PMID- 14527103 TI - In search of a course design and teaching methods to improve critical thinking skills. AB - This article describes a unique course design to teach critical thinking skills to students in formal academic degree programs preparing to become healthcare administrators. The course and its experimental design were motivated by several factors. First, intelligent and competent problem solving and decision making, most would agree, are directly dependent on critical thinking proficiency. Therefore, elevating students' critical-thinking competence will inherently improve their problem-solving and decision-making ability. Second, problem solving and decision making consumes the majority of an administrator's time. And finally, a review of AUPHA's catalog descriptions of healthcare graduate curriculums listrelatively few critical-thinking courses compared to the time administrators devote to it. PMID- 14527104 TI - Validation of data collection for the HEDIS performance measure on chlamydia screening in an MCO. AB - OBJECTIVE: To determine the validity of calculating the chlamydia Health Plan Employer Data and Information Set (HEDIS) measure using administrative data available in a mixed-model managed care organization (MCO). STUDY DESIGN: Retrospective cohort study. METHODS: A review of International Classification of Diseases, Ninth Revision (ICD-9), Current Procedural Termin-ology (CPT), Healthcare Common Procedure Coding System (HCPCS), and National Drug Code codes and electronic laboratory files in 1998 and a medical chart review to validate sexual activity and chlamydia testing codes specified by the National Committee for Quality Assurance (NCQA) in 1999 for the chlamydia HEDIS 2000 measure. RESULTS: Fewer than 25% of female enrollees with laboratory evidence of a chlamydia test had a CPT code for chlamydia testing as specified by the NCQA. Non pathogen-specific test codes instead of NCQA-specified codes were used in 1998 to code chlamydia tests. By incorporating electronic laboratory data into the automated claims-generating process, all chlamydia tests performed at staff-model clinics were coded. Use of pharmacy dispensing data to identify contraceptive prescriptions increased the proportion of enrollees classified as sexually active by 4% to 5% vs documentation of sexual activity using ICD-9, CPT, and HCPCS codes only. CONCLUSIONS: The MCO quality assurance specialists examining chlamydia testing rates under HEDIS may want to evaluate chlamydia testing coding practices in their MCOs to determine whether simple changes in coding practices may present a more accurate picture of actual testing practices. The proportion of female enrollees classified as sexually active using different data available in the staff and network models varied only slightly. PMID- 14527105 TI - Improvements in access to care for HIV and AIDS in a statewide Medicaid managed care system. AB - BACKGROUND: Some experimental Medicaid managed care systems have expanded eligibility criteria for chronically ill persons, but these systems' impact on access to care remains unknown. OBJECTIVE: To determine whether initiating a statewide Medicaid managed care system (TennCare) guaranteeing universal access for persons living with HIV or AIDS (PLWHs) increased their enrollment in public sector insurance. DESIGN, SETTING, AND PARTICIPANTS: A retrospective longitudinal descriptive analysis of trends in population characteristics during the study period was performed. The study population included all PLWHs in Tennessee (1992 1997) identified by the State Health Department. These data linked with Medicaid/TennCare enrollment files identified percentages of Tennessee's HIV/AIDS population enrolled in Medicaid (1993) or TennCare (1994-1997) and eligi bility/demographics changes during program initiation. MAIN OUTCOME MEASURE: Annual percentage of PLWHs enrolled in Medicaid/TennCare. RESULTS: Absolute numbers of PLWHs served by Medicaid/TennCare increased 475% from 1992 (n = 593) to 1997 (n = 2818). Similar increases in Tennessee's overall HIV-positive population occurred. Percentages of PLWHs enrolled in Medicaid/TennCare increased (1993 to 1997): HIV (28% to 34%) and AIDS (32% to 44%). The largest percentage of PLWHs added to the program were uninsured/uninsurable. CONCLUSIONS: Absolute numbers of PLWHs covered by Medicaid/TennCare substantially increased. Percentages of PLWHs covered increased more modestly, partly owing to large increases in overall numbers of HIV-positive Tennesseans during the study period. Increases in coverage were greatest for the AIDS population. Tennessee's broad expansion of eligibility for PLWHs resulted in improved access, but did not result in enrollment of most PLWHs. States contemplating similar Medicaid expansions should not expect all PLWHs to crowd into public sector insurance programs. PMID- 14527107 TI - The effect of HMO competition on gatekeeping, usual source of care, and evaluations of physician thoroughness. AB - OBJECTIVES: To examine the effects of HMO enrollment and HMO competition on evaluations of physician thoroughness through their effects on gatekeeping and having a usual source of care and to determine whether the effects of HMO competition spill over to individuals not enrolled in HMOs and whether these effects differ in those enrolled vs not enrolled in HMOs. STUDY SAMPLE: A nationally representative sample of 27 441 adults from the household component of the Community Tracking Study-Round 1 (July, 1996, through July, 1997). STUDY DESIGN: A retrospective econometric analysis of Community Tracking Study data merged with measures of HMO competition. METHODS: Gatekeeping was regressed on HMO enrollment, HMO competition, and control variables using ordered logistic regression. Usual source of care was regressed on gatekeeping, HMO enrollment, HMO competition, and control variables using logistic regression. Evaluation of physician thoroughness was regressed on gatekeeping, usual source of care, HMO enrollment, HMO competition, and control variables using multivariate regression. RESULTS: HMO competition increases use of gatekeeping and gatekeeping increases having a usual source of care for all individuals. For HMO enrollees, HMO competition increases having a usual source of care, whereas for those not in HMOs, it decreases having a usual source of care. For all individuals, having a usual source of care increases evaluation of physician thoroughness. For those in HMOs, gatekeeping decreases evaluation of physician thoroughness. CONCLUSIONS: For HMO enrollees, the overall effect of HMO competition is to increase evaluations of physician thoroughness. For those not in HMOs, although there are HMO competition spillover effects, they are offsetting, resulting in no overall effect of HMO competition on evaluations of physician thoroughness. PMID- 14527106 TI - Effects of protocol-driven care versus usual outpatient clinic care on survival rates in patients with type 2 diabetes. AB - OBJECTIVE: To determine whether clinical outcomes in patients with type 2 diabetes were improved by protocol-driven care in a Diabetes Centre compared with usual outpatient care. STUDY DESIGN: Descriptive analysis of a prospective cohort. PATIENTS AND METHODS: During a median 7-year observational period, 91 patients with type 2 diabetes and no cardiovascular or renal complications were monitored by a nurse and a diabetologist in a clinical trial setting according to a structured protocol. Another 81 patients with comparable clinical characteristics were monitored by generalists at the medical clinic in the same hospital. Clinical end points, defined as death and cardiovascular and renal events, were evaluated in 1997 by review of case records. RESULTS: Patients receiving structured care had lower mortality (relative risk [RR] = 0.21; 95% confidence interval [CI] = 0.07, 0.65; P = .006) than the usual-care group, as well as a lower incidence of combined clinical end points (RR = 0.43; 95% CI = 0.22, 0.84; P = .01). In the usual-care group, patients who had no monitoring of glycosylated hemoglobin or plasma lipid levels during the entire observational period (8.6%) had a 14.6-fold (P < .01) and 15.7-fold (P < .01) increased risk of death and combined clinical end points, respectively, compared with those who had at least one measurement (60.5%). CONCLUSION: Management by protocol-driven care model improved survival and clinical outcomes in patients with type 2 diabetes. Definitive studies are required to confirm these findings and compare the cost effectiveness of these care models. PMID- 14527109 TI - A case study on rheumatoid arthritis. AB - At a time when many managed care organizations increasingly shift costs to patients through tiered formularies and widening copay differentials, biologic agents represent a significant clinical and financial challenge unlikely to be managed optimally with tiered formularies and greater patient cost sharing. The information discussed in this article is intended for healthcare professionals involved with rheumatoid arthritis therapy, including but not limited to physicians in both the inpatient and outpatient setting, and for other managed care professionals, including medical directors, pharmacy directors, long-term care decision makers, nurses, pharmacists, and case managers. PMID- 14527108 TI - The public's opinions of physicians: do perceived choice and exercised choice matter? AB - OBJECTIVE: To assess whether the public's opinions of physicians are affected by one's perceived ability to choose providers and by past experience with switching providers or health plans. STUDY DESIGN: Cross-sectional survey of the American general public in 1998. METHODS: Multivariate quantitative analyses of opinions according to respondents' perceived ability to choose providers, history of exercising choice of providers or plans, and health status. Exploratory factor analyses resulted in 4 scales of opinions: trusting personal physician, positive opinions of physicians as a group, negative opinions of physicians as a group, and concerns about the influence of managed care on personal physicians' practice patterns. RESULTS: Although 75% (879/1172) of the sample perceived that they could use any physician they wanted, only 20% had ever switched health providers or plans owing to dissatisfaction. Perceived lack of choice is associated with lower levels of trust in personal physicians and with higher levels of concern about the influence of managed care on personal physicians. Individuals who have not switched in the past are more positive about physicians as a group and are less concerned about the influence of managed care. Health status affected opinions significantly. Educational attainment is negatively associated with opinions. CONCLUSIONS: The public's opinions of physicians are significantly associated with one's perceived ability to choose any provider, past experience with switching health providers or plans, health status, and education. Ensuring consumer choice of providers and targeting those who have switched in the past and the sick could potentially improve the public's opinions. Meaningful provider quality information is necessary to ensure choices that can enhance consumer welfare. PMID- 14527110 TI - The functional repertoire of the hand and its application to assessment. AB - Functional assessment of the hand typically involves evaluation of proficiency using a test battery rather than identifying the range of hand skills available to a client. Establishing a clients hand function by reference to the movement repertoire of his or her fully-functioning hand offers numerous benefits for clinicians and researchers. A functional repertoire model of the hand is proposed as a framework for guiding assessment and therapy. The model identifies four main components relating to hand function: personal constraints, hand roles, hand actions, and task parameters. The model provides a common language for assessment, program development, and research across populations. Application here is to hand-injured clients, but the framework is equally valuable for hand function assessment of developmentally and physically challenged individuals, persons with neuromuscular disorders, or geriatric populations. PMID- 14527111 TI - Outcomes in hand rehabilitation using occupational therapy services. AB - OBJECTIVE: The purpose of this study was to measure functional outcomes after outpatient occupational therapy for clients who had upper-extremity injury and surgery or both. METHODS: A sample of 33 clients referred to occupational therapy outpatient intervention was recruited from five clinics in Ohio. The Canadian Occupational Performance Measure (COPM) was used to guide the occupational therapy sessions and to measure outcomes. The Disability of Arm, Shoulder, and Hand (DASH) questionnaire and the Short Form 36 (SF-36) were also administered to the clients pre and post 6 to 8 weeks of hand rehabilitation services. Two to three months after discharge, the clients responded by the telephone to the Community Integration Questionnaire. RESULTS: The clients received a mean of 13 hours of outpatient occupational therapy services and received no other rehabilitation service. Functional performance gains following 6 to 8 weeks of services were significant (COPM; t(32) = -11.5, p < .001; t(32) = -11.31, p < .001; DASH; t(32) = 9.22, p < .001; t(32) = -9.02, p < .001). Effect sizes on the COPM and DASH ranged between 1.43 and 2.52. Progress in the clients' goals was moderately correlated to progress in functional measures. CONCLUSION: Clients with upper-extremity injury or surgery made strong, positive gains in functional measures following client-centered occupational therapy services. The COPM was the most sensitive to client change, followed by the DASH, and then the SF-36. PMID- 14527112 TI - Weighted wrist cuffs for tremor reduction during eating in adults with static brain lesions. AB - OBJECTIVE: This study examined whether weighting the forearm during feeding decreased tremors and increased functional feeding in adults with intention tremor caused by static brain lesions. METHOD: Five individuals with various diagnoses, ages 30-81, were videotaped during 8 or 16 meal sessions, alternating treatment and control conditions within each meal. In this single-case design, treatment consisted of application of a weighted fabric wrist cuff and the baseline (control) condition employed an identical cuff with the weights removed. Dependent variables studied were time to acquire and deliver a bite, grams of food eaten, number of times food was spilled, number of times a compensatory technique was used, participant self-rating, and investigator rating of the severity of the tremor. RESULTS: All five participants demonstrated improvement during treatment in one or more of the dependent variables. t Tests of the means of baseline and treatment half-sessions incorporating conservative control of Type I error revealed the following statistically significant improvements under the weighted condition: Participants 3, 4, and 5 took less time to acquire a bite; Participants 4 and 5 made fewer spills; Participants 3 and 5 showed a diminished tremor. There were no statistically significant decreases in function on any variable for any participants during the weighted condition. CONCLUSION: The application of weight to the wrist of a person with upper-extremity tremor is accompanied by some functional improvement in self-feeding for some individuals. The size of benefit seems to be sensitive to the amount of weight used. PMID- 14527114 TI - Spirituality as experienced by occupational therapists in practice. AB - Despite considerable literature describing the potential place of spirituality in occupational therapy, surveys repeatedly demonstrate that therapists are uncomfortable with this concept in practice. To gain a better understanding of how spirituality might inform practice, we interviewed eight occupational therapists who stated that they considered spirituality while working with patients. Participants defined spirituality as one's beliefs about the world and one's place in it and how one lives out these beliefs, through reflection and conscious actions. Four themes of consideration of spirituality in practice emerged. In the first, addressing religious concerns, therapists dealt with patients' religious questions and issues. In the second, addressing suffering, therapists assisted patients to deal with their feelings related to loss and pain, attempted to relieve patients' distress and helped patients move towards increased functioning. In the third, encouraging the self, therapists worked to assist patients to acknowledge their own worth and to use their unique gifts and interests. In the fourth, growing as a person, therapists themselves were transformed as a result of the therapeutic encounter. This study represents an early attempt to determine what occupational therapists who consider spirituality actually do in practice. Future research should go deeper into the experience of such therapists, to gain a richer understanding of the phenomenon. PMID- 14527113 TI - The immediate and short-term effects of a wrist extension orthosis on upper extremity kinematics and range of shoulder motion. AB - OBJECTIVE: The purpose of this study was to investigate upper-extremity kinematics and range of shoulder motion while wearing a wrist extension orthosis compared to the free hand at initial testing and after a week's wear. METHOD: A convenience sample of healthy women, ages 20-50 years, performed a stacking task and a pouring task, freehanded and splinted, at two times, 1 week apart in this counterbalanced, repeated measures design. A Motion Analysis 3-D system measured quality of movement and range of motion variables. A 2 by 2 analysis of variance (splinted vs. freehanded by time of testing) was used to analyze each variable. RESULTS: In both tasks, participants tended to move more slowly (p = 0.005) with less direct movements (p < .001) when wearing the orthosis. In stacking, splinted movements were also less smooth than when free-handed (p < .0001). When splinted both tasks required more shoulder abduction (p < .0001) and pouring required more shoulder flexion (p < .0001). Participants demonstrated several improvements in quality of movement and amount of shoulder motion used after a weeks' orthotic wear, indicating some adaptation to the orthosis with use. CONCLUSION: In this study, a wrist extension orthosis interfered with the quality of upper-extremity movement and required more range of shoulder movement compared to the free hand. Future research is needed to determine whether persons with upper-extremity conditions (i.e., arthritis, carpal tunnel syndrome) respond similarly to wrist orthoses. Therapists should consider discussing with patients how their wrist splint could affect their hand performance and help patients learn techniques to reduce stress on their proximal joints. PMID- 14527115 TI - Classroom seating for children with attention deficit hyperactivity disorder: therapy balls versus chairs. AB - OBJECTIVE: The purpose of this study was to investigate the effects of therapy balls as seating on in-seat behavior and legible word productivity of students with attention deficit hyperactivity disorder (ADHD). Additionally, social validity was assessed to evaluate teacher and student opinions regarding the intervention. METHOD: A single subject, A-B-A-B interrupted time series design was employed across 3 students (2 males, 1 female) with ADHD. The study was conducted in a 4th grade inclusive classroom during daily language arts. During phases 1 and 3, the 3 participants and all other class members sat on chairs (in seat on chair); during phases 2 and 4, everyone sat on therapy balls (in-seat on ball). Dependent variables were in-seat behavior and legible word productivity. Data were graphed and visually analyzed for differences between phases. RESULTS: Results demonstrated increases in in-seat behavior and legible word productivity for the students with ADHD when seated on therapy balls. Social validity findings indicated that generally the teacher and students preferred therapy balls. CONCLUSION: This study provides evidence that use of therapy balls for students with ADHD may facilitate in-seat behavior and legible word productivity. PMID- 14527117 TI - Fine motor activities in Head Start and kindergarten classrooms. AB - OBJECTIVE: The purpose of this study was to describe and compare the fine motor activities in Head Start and kindergarten classrooms in order to open a dialogue between the two contexts about the fine motor activities children in preschool will face in kindergarten. METHODS: Children in 10 Head Start and 10 kindergarten classrooms were observed for 1 day each. Time spent in activities was categorized into four groups: fine motor activities with no academic purpose, fine motor activities with academic purpose, academic activities with no fine motor component, and nonacademic activities with no fine motor component. Percentages of time were calculated and t tests were used for comparison between contexts. RESULTS: Children in Head Start spent 27%-46% (mean of 37%) of the in-class day in all fine motor activities whereas children in kindergarten spent 36%-66% (mean of 46%). For children in kindergarten, 42% of total fine motor activity time involved paper and pencil activities. In contrast, children in Head Start spent 10% of total fine motor activity time in paper and pencil activities. CONCLUSION: The higher mean percent of time spent in fine motor activities in kindergarten classrooms suggests a developmentally appropriate increase in fine motor demands. The percent of paper and pencil activity time the children engaged in substantially increased from Head Start to kindergarten. The findings describe a difference between the two environments, informing Head Start of the fine motor demands their graduates face in kindergarten. PMID- 14527116 TI - Occupational therapy effects on visual-motor skills in preschool children. AB - OBJECTIVE: The purpose of this study was to evaluate the assumption that preschool children who receive occupational therapy will demonstrate significant improvement in their visual-motor skills as measured on the Developmental Test of Visual-Motor Integration (VMI) and the two supplemental Visual Perception and Motor Coordination tests. METHOD: Preschool children with developmental delays (n = 12) received occupational therapy a minimum of one individual 30-minute session, and one group 30-minute session per week for 1 school year. Their performance was compared to that of two control groups; preschool students without disabilities who received occupational therapy (n = 16) for one 30-minute group session per week and students without disabilities (n = 15) who received no occupational therapy. The VMI and two supplemental tests were administered three times to each student, at the beginning, middle, and end of school year. RESULTS: Planned comparison tests showed that students with developmental delays demonstrated statistically significant improvement in visual-motor skills and developed skills at a rate faster than expected when compared to typically developing peers on the VMI. The effect size for preschool students without disabilities who received occupational therapy exceeded the effect size for the VMI and Visual Perception supplemental test for the preschool students without disabilities who received no therapy, although, the difference in the post-test performance of these two groups was not statistically significant. DISCUSSION: The results of this study demonstrate that intervention, including occupational therapy, can effectively improve visual-motor skills in preschool-aged children. PMID- 14527118 TI - Daily time use as a measure of community adjustment for persons served by assertive community treatment teams. AB - PURPOSE: The purpose of the study was to examine daily time use of clients of Assertive Community Treatment (ACT) as a measure of their community adjustment and well-being. The actual daily time use of ACT clients in the four categories of personal care, productivity, leisure, and sleep were compared to the data for Canadian population norms. METHOD: Daily time use data were collected from 27 adult clients from two Assertive Community Treatment Teams in southeastern Ontario using recall time diaries of two weekdays. The data were coded using the Statistics Canada (1999) coding scheme. Descriptive statistics were used to determine time in the major categories of time use and z scores were used to compare the study sample to the adult Canadian population. The percentages of time spent in specific subcategories of activity were also compared. RESULTS: The results indicated an imbalance in occupation with time use dominated by leisure and sleep activities. Study participants spent significantly more time in passive leisure compared to active leisure and socialization. CONCLUSION: The activity patterns of ACT clients were not consistent with those associated with community adjustment, health, and well-being. Occupational therapists working in ACT are in a good position to contribute to the literature regarding occupational performance and mental illness and to lead ACT teams in discussions and practices that may promote health through activity. PMID- 14527119 TI - A comparison of consultative model and direct-indirect intervention with preschoolers. AB - OBJECTIVE: The purpose of this study was to compare the effectiveness of a consultative model of intervention with that of direct-indirect intervention for meeting goals of preschool students with mild motor delays. METHOD: Two occupational therapists provided consultation to teams working with 11 children 1 day a week for all 40 weeks of the school year. A different occupational therapist provided direct-indirect treatment to another 9 children for a full week every 3 weeks for the 40 weeks of the school year. Data for both groups used in this study were kept for the last 4 months at the end of the school year or as the children appeared to be nearing completion of their goals of the study. All therapists met with their teams twice a month to plan and review progress. All parents carried out activities with their children at home. Goal Attainment Scaling was used to examine progress. RESULTS: No statistically significant differences were found between the two types of intervention (t = .359; df = 18; p = .724). CONCLUSION: The study suggests that a consultative model and a direct indirect model are equally effective in meeting objectives for preschoolers with relatively mild motor impairments. PMID- 14527120 TI - Adult norms for a commercially available Nine Hole Peg Test for finger dexterity. AB - The Nine Hole Peg Test is commonly used by occupational therapists as a simple, quick assessment for finger dexterity. The purpose of this study was to evaluate the interrater and test-retest reliability of the commercially available Smith & Nephew Rehabilitation Division version of the Nine Hole Peg Test, and to establish new adult norms for the Nine Hole Peg Test for finger dexterity utilizing this particular version. Two of the researchers established interrater and test-retest reliability by evaluating 25 occupational therapy student volunteers. Seven hundred and three subjects, ranging in age from 21 to 71+ years, were tested to establish norms, using the standard protocol. Results showed high interrater reliability and only moderate test-retest reliability. Scores obtained by using the commercially available version were not statistically different from previously published norms (Mathiowetz, Weber, Kashman, & Volland, 1985). This study supports the original norms and further assists occupational therapists to evaluate dexterity accurately. PMID- 14527121 TI - Interrater and test-retest reliability of the Movement Assessment Battery for Chinese preschool children. AB - OBJECTIVE: The aim of this paper was to check on the reliability of the Movement Assessment Battery for Children (Movement ABC) in preparation for its standardization in Hong Kong. Interrater and test-retest reliability are reported for Age Band One, designed for use with children ages four to six. METHOD: Interrater reliability of the Movement ABC was estimated using two trained observers with 79 children. Test-retest reliability was assessed using 75 children who were tested twice by the same tester over a 2- to 3-week interval. RESULTS. Agreement between testers was good with a mean intraclass correlation of 0.96 across items. A value of 0.77 was obtained for test-retest reliability. CONCLUSION: These results support the use of this component of the Movement ABC in Hong Kong. PMID- 14527122 TI - Self-image, adolescence, and disability. AB - The purpose of the present study was to examine the self-image of a group of young persons with cerebral palsy. The respondents (3 girls and 4 boys, ages 12 to 17 years) were interviewed and asked to complete a personality inventory. The results from the personality inventory revealed that the respondents viewed themselves in a very positive manner and rated markedly higher than norm groups. This positive view corresponded well with the findings of the interview. The following conclusions are made: Further studies on self-image and the psychosocial development of adolescents with disabilities should (a) focus on the social interaction outside their immediate families, and (b) continue to use and develop methods where these adolescents can give voice to their own experiences and opinions. PMID- 14527123 TI - Toward culturally relevant epistemologies in occupational therapy. PMID- 14527124 TI - The role of professional expertise in evidence-based occupational therapy. PMID- 14527125 TI - Integration of universal design of instruction in occupational therapy professional education: responding to student diversity. AB - Application of the seven principles of universal design of instruction in occupational therapy education is intended to meet the diverse learning needs of students and provide full access to educational experiences without disadvantaging some students (Bowe, 2000; Burgstahler, 2001; McGuire & Scott, 2002). The intended outcome is success for all students. Further, universal design of instruction is congruent with the occupational therapy core value of mutual respect, whereby... We treat people fairly and equitably. We acknowledge and support others regardless of differences. We appreciate their qualities, capabilities and contributions (American Occupational Therapy Association, 2003). While the benefits of universal design of instruction in occupational therapy curricula appear to be extensive, a caveat must be included. Use of any instructional design tool is legitimized when grounded in research. Though literature in multiple disciplines is expanding related to universal design of instruction, we need research specific to occupational therapy education. PMID- 14527126 TI - Inuit health in Greenland: a population survey of life style and disease in Greenland and among Inuit living in Denmark. AB - During 1997-2001 a population survey was carried out amongst Greenland Inuit living in Denmark and West Greenland (Nuuk, Sisimiut, Qasigiannguit and four villages in Uummannaq municipality). Data collection comprised an interview, a questionnaire, clinical examinations and sampling of biological specimens (blood, urine, subcutaneous fat tissue). The clinical examinations included anthropometric measurements, an oral glucose tolerance test, ECG, ultrasound of thyroid gland and carotid arteries, a skin prick test, and lung function. The data collection areas in Greenland ranged from the westernized capital of Nuuk (pop. app. 13,000) to small fishing and hunting villages (pop. app. 250). A total of 4,162 persons aged 18+ participated in the study; clinical examinations were performed on 2,056 of these, 739 from Denmark and 1317 from Greenland. Some of the above mentioned procedures were performed on a subset of the participants. The participation rate was 62%. We provide an overview of the background of the study and a detailed description of the methods employed for the data collection. A set of standard tables are provided for the indigenous population of Greenland. These cover statistics for selected variables by gender and ten-year age groups. PMID- 14527127 TI - Hypothalamic neuronal networks and feeding-related peptides involved in the regulation of feeding. AB - The hypothalamus is a region of the brain that plays a critical role in feeding regulation. It has been revealed by various physiological experiments that the feeding-regulating center is confined to the ventromedial hypothalamus, lateral hypothalamus (LH) and arcuate nucleus (ARC). Many kinds of neurons in these areas of the hypothalamus express factors such as melanin-concentrating hormone (MCH), neuropeptide Y (NPY), proopiomelanocortin (POMC), orexin (OX) and ghrelin, which have been implicated in feeding regulation. In tissues of the periphery, two critical factors involved in feeding regulation, leptin and ghrelin, have been identified. Both hormone peptides are secreted mainly from adipose and stomach tissue, respectively, and are considered to function via their receptors mainly through several hypothalamic nuclei that play important roles in the regulation of appetite. The present review looks mainly at the functional significance of feeding-regulation factors, such as those described above, and the humoral and neuronal interactions among these compounds in the hypothalamus by drawing on published reports of morphological and physiological analyses. Immunohistochemical and in situ hybridization experiments indicate that both leptin and ghrelin receptors are distributed in the hypothalamus and that there are reciprocal interactions between MCH and OX neurons in the LH. Morphological and physiological studies on single living cells isolated from fresh rat hypothalamus or with receptor agonist and antagonist combined with immunohistochemisry clearly demonstrate that both leptin and OX reciprocally regulate NPY- and POMC-containing neurons in the ARC and that ghrelin may regulate feeding status independently through direct OX and NPY pathways. In this way, cross-talking systems in the hypothalamus play a role in determining feeding states. PMID- 14527128 TI - Three gonadotropin-releasing hormone neuronal groups with special reference to teleosts. AB - Gonadotropin-releasing hormone (GnRH) is a decapeptide, which has been isolated from the hypothalamus as a releasing hormone of gonadotropins from the pituitary. However, subsequent morphological studies have demonstrated the presence of multiple GnRH neuronal groups outside the hypothalamus and preoptic area. In most vertebrate lineages studied to date, GnRH neuronal groups are present along the terminal nerve and in the midbrain tegmentum, in addition to a population in the preoptico-hypothalamic areas. The presence of GnRH fibers in extrahypothalamic areas has also been demonstrated, indicating a significance for GnRH neurons in functions other than those that are purely hypophysiotropic. Among vertebrate lineages, GnRH neurons have been most extensively studied in teleost fish through morphological, electrophysiological, behavioral and molecular approaches. To date, studies on differential roles of GnRH neuronal groups have been mostly restricted to teleosts. In the present review, the anatomy and functions of each GnRH neuronal group are reconsidered, based mainly on knowledge from teleosts. Recent findings in teleosts indicate that the preoptico-hypothalamic GnRH neurons are hypophysiotropic and that GnRH neurons of the terminal nerve and midbrain tegmentum regulate neural activities in various regions, including extrahypothalamic areas. The latter populations presumably serve as neuromodulatory systems to control aspects of neural functions such as reproductive behavior. Similar functional differentiation may be generalized to other vertebrate lineages as well. PMID- 14527129 TI - Poly lactic acid--caprolactone copolymer tube with a denatured skeletal muscle segment inside as a guide for peripheral nerve regeneration: a morphological and electrophysiological evaluation of the regenerated nerves. AB - A biodegradable copolymer of poly L-lactic acid and epsilon-caprolactone (PLAC) was manufactured into a tube, in which a denatured skeletal muscle segment was placed longitudinally. This model tube was implanted as a guide to promote nerve regeneration across a 5 cm gap in the rabbit sciatic nerve. Five months after implantation, good nerve regeneration was found throughout the graft and in the distal host nerve. The population (29.6/16 x 10(2) microm(2)) of regenerated nerves in the graft was higher than that of the contralateral normal sciatic nerve (18.0/16 x 10(2) microm(2)). Regenerated nerve fibers extended to the distal host nerve. The number of myelinated fibers was 13.7/16 x 10(2) microm(2) at a level 1.5 cm from the distal suture. The diameters (below 2 microm) of most regenerated myelinated (nerves in the graft and in the distal host nerve were much smaller than those (6-8 microm) of normal nerves. Electrophysiological evaluation showed that the hindlimb muscle (gastrocnemius) was innervated by motor nerves in all animals 5 months after implantation. These results indicate that the PLAC tube with a denatured muscle segment inside provided good conditions for nerve fiber regrowth. The PLAC tube is thought to protect the denatured muscle segment from rapid dissociation in the host tissue. PMID- 14527130 TI - Histological investigation of rabbit ligamentum flavum with special reference to differences in spinal levels. AB - The structure of the ligamentum flavum has yet to be fully elucidated and no studies have investigated fine structural differences at different spinal levels in any animals. The aim of the present study was to clarify structural differences in the ligamentum flavum at different spinal levels (cervical: C3/4 and C5/6; upper thoracic: T2/3; lower thoracic: T9/10; lumbar: L3/4) using light and electron microscopy of rabbit specimens. Light microscopy using resorcin fuchsin staining revealed that the distribution of elastic fibers was diffuse in the cervical and upper thoracic regions, but was generally dense in the lower thoracic and lumbar regions. Electron microscopy demonstrated that the cervical and upper thoracic regions were rich in collagen fibers. Conversely, the lower thoracic and lumbar regions were rich in elastic fibers. Quantitative image analyses displayed thick elastic fibers in the lower thoracic and lumbar regions, with high area ratios. Radiographic examinations revealed that ranges of motion were large at the cervical region, but small at the lower thoracic and lumbar regions. These findings suggest that structure of the ligamentum flavum varies at different spinal levels with respect to differences in motion. PMID- 14527131 TI - Three-dimensional architecture of the intrinsic tongue muscles, particularly the longitudinal muscle, by the chemical-maceration method. AB - Muscle bundles of the transverse and vertical muscles of the tongue become flat when they enter the longitudinal muscle layers of the tongue, where they form a tunnel-like structure that surrounds the longitudinal muscle of the tongue. However, the three-dimensional architecture of longitudinal muscle fibers of the tongue has not been clarified. In the present study, we evaluated the function of the intrinsic muscles of the tongue by studying the three-dimensional architecture of the longitudinal muscle. Muscle bundles of the longitudinal muscle of the anterior part of a rabbit's tongue were exposed by the chemical maceration and modified chemical-maceration methods and examined by scanning electron microscopy. In the longitudinal muscle of the tongue, muscle bundles running in the anteroposterior direction were arranged at regular intervals. These muscle bundles bifurcated or ramified at a sharp angle at each level from the superficial layer to the deep layer and joined or fused with adjacent muscle bundles. In addition, these ramified muscle bundles ran obliquely into shallower or deeper layers of the muscle, as well as in the same plane. Consequently, the longitudinal muscle of the tongue as a whole had a three-dimensional mesh-like structure. The transverse and vertical muscles of the tongue entered this mesh like structure of muscle bundles of the longitudinal muscle as flat muscle bundles. The transverse and vertical muscles showed no ramification in the center of the tongue, where there is no longitudinal muscle. These results suggest that the three intrinsic muscles of the tongue are interlaced with one another and are bound tightly in the longitudinal muscle. This structure may enable the dorsum of the tongue to harden for pressing food during mastication and shifting the food posteriorly for swallowing. PMID- 14527132 TI - Superficial brachial artery crossing over the ulnar and median nerves from posterior to anterior: embryological significance. AB - A rare variation of the superficial brachial artery was found in the right arm of an 82-year-old male cadaver in student dissection practice. In this case, the axillary artery passed normally between the medial and lateral roots of the median nerve and then bifurcated into a large superficial brachial artery and the deep brachial artery (A. brachialis profunda). The superficial brachial artery passed medially to the ulnar nerve and then crossed over to take a lateral course to the median nerve at the midpoint of the upper arm. It finally divided into the radial and ulnar arteries at the cubital fossa. The deep brachial artery terminated as the inferior ulnar collateral artery. These findings indicate that the present variant was a well-developed medial type of the superficial brachial artery that gave off the ulnar and radial arteries. The anatomical and embryological significance of the findings are discussed. PMID- 14527133 TI - Preliminary report on the correlations among pineal concretions, prostatic calculi and age in human adult males. AB - By using quantitative image analysis of soft X-ray photographs on the bulk of extracted pineal glands and prostates, we made a preliminary investigation into the correlations among pineal concretions (% by mass), prostatic calculi (% by mass) and age (years) in 40 human adult males, ranging in age from 31 to 95 years (mean (+/-SD) 69.9 +/- 15.2 years), who died and underwent the routine dissection course. The mass concentrations of pineal concretions and prostatic calculi were 17.68 +/- 13.56% (range 0-51.34%) and 0.93 +/- 1.31% (range 0-5.82%), respectively. There was no correlation between the mass concentration of pineal concretions and aging (r = 0.03; P < 1.0). There was no correlation between mass concentration of prostatic calculi and aging (r = 0.28; P < 0.5). No pineal concretions and no prostatic calculi were observed in seven and 10 cases, respectively; in addition, in one case, neither-concretions nor calculi were seen. From such data and from the previously reported suggestion on the counteracting functions between the pineal gland and prostate, a negative correlation between the mass concentrations of pineal concretions and prostatic calculi was expected. This was certainly obtained, but the correlation was low (r = -0.39; P < 0.05). Such a low correlation and no correlations between the concentrations of pineal concretions and aging or between prostatic calculi and aging may have been caused by the examination of relatively older humans. Therefore, further investigations using a number of pair samples collected from males including younger age generations will be necessary. PMID- 14527134 TI - Abnormal anterior belly of the digastric muscle: a proposal for the classification of abnormalities. AB - We recognized an abnormal anterior belly of the digastric muscle in an 83-year old male cadaver. Three muscle bundles were observed on the left anterior belly: (i) attached to the left digastric fossa; (ii) attached to the right digastric fossa; and (ii) attached to the raphe of the mylohyoid muscle. Four muscle bundles were recognized on the right anterior belly: (i) attached to raphe of the mylohyoid muscle; (ii, iii) attached to the exterior surface on the base of the mandible from the raphe of the mylohyoid muscle; and (iv) attached to the interior surface on the base of the mandible from the raphe of the mylohyoid muscle. The raphe of the mylohyoid muscle was curved significantly to right and the four abnormal bundles found on the right anterior belly (see above) were attached to its curved point. PMID- 14527135 TI - Case studies from morning report: librarians' role in helping residents find evidence-based clinical information. AB - In primary care specialties, Morning Report is a traditional vehicle for expanding medical residents' training in diagnosis and treatment. At one academic medical center, residents and faculty in the Department of Family and Community Medicine use case-based teaching, centered around planning and reviewing patient management, to review intriguing cases from patient encounters in the department's hospital service. Seeking to improve the level of evidence-based information exchanged at Morning Report, department leaders invited reference librarians from the health sciences library to attend weekly Morning Report. The librarians saw this as an opportunity not only to improve residents' information seeking skills, but also to improve librarians' teaching skills and understanding of the needs of users in clinical settings. This paper describes the evolution of librarians' involvement in Morning Report, examples of the kinds of contributions librarians have made in this setting, and changes made in Morning Report sessions to facilitate this activity. PMID- 14527136 TI - Making sense of liver enzymes, liver function tests, and hepatobiliary disorders at the reference desk. AB - This paper discusses concepts and terminology of some aspects of the laboratory diagnosis of liver (hepatic) disease and biliary tract disease (hepatobiliary disease) as it relates to medical reference work. Details of anatomic, biochemical, and pathologic processes are not discussed. Knowledge of the specific terminology involved in this area may help to ensure a good approach to developing prudent strategies for database searching of the medical literature and therefore is reviewed. MeSH and EMBASE thesauri terms are discussed and textword synonyms are presented that provide tools for thorough searching techniques. Commonly used medical jargon for this area is also explained. Examples of specific search strategies are illustrated. PMID- 14527137 TI - Complementary and alternative medicine as represented in the HIV/AIDS body of knowledge: a bibliometric analysis. AB - Although the HIV/AIDS epidemic began more than twenty years ago, there still is no cure for the disease and no vaccine to prevent infection. As with the general population, individuals with HIV/AIDS have sought care using a variety of traditional and nontraditional approaches. The popularity of complementary and alternative interventions among the HIV/AIDS community continues. To understand better the distribution of the HIV/AIDS body of knowledge concerning complementary and alternative medicine (CAM), this study sought to (1) examine the literature specific to the use of complementary and alternative medicine where HIV/AIDS is concerned to determine publication patterns; (2) determine the degree of overlap among bibliographic citation databases that index the literature concerning the use of CAM practices in treating HIV/AIDS; and (3) facilitate access to this body of literature. PMID- 14527138 TI - Natural Standard--new integrative medicine database. AB - Natural Standard is a new and extremely easy-to-use database consisting of comprehensive evidence-based medicine monographs, and also news brief's on the subject of complementary/alternative medicine therapies. It does not yet contain a huge amount of material, but it is updated frequently, and new information is continuously added. The focus is on the therapeutic "bottom line" of treatment safety and effectiveness. Dosing, Precautions/Adverse Effects, Pregnancy/Lactation Data, and Interactions information is all clearly displayed along with a "Review of the Evidence" table and a discussion summarizing the overall therapeutic value of the treatment modality. PMID- 14527139 TI - Non-governmental health statistics on the World Wide Web. AB - Health statistics are an important decision-making resource for professionals and consumers alike. Professional use of statistics is wide ranging and often includes legislative decision making, medical research, grant writing, and population analysis. Locating needed or even helpful statistics to make these decisions or analyses can seem overwhelming. Organizations advocating for the treatment or assessment for particular diseases and health conditions are a prime resource for needed statistics. This column describes links and provides summaries to organizations that offer statistical resources for many of the leading causes of disability and death; it is the second of two columns devoted to finding health statistics on the Web. A helpful search strategy is presented which includes the utilization of evaluative criteria to seek legitimate and credible information. PMID- 14527140 TI - The Maine electronic document delivery project: a cooperative project of Maine hospital libraries and the NN/LM New England Region. AB - The article discusses an Ariel document delivery project with seven Maine hospital libraries, the University of Massachusetts Medical School Library (UMass), and the New England Regional office of the National Network of Libraries of Medicine. Funding was awarded to six network members to purchase equipment or Ariel software. UMass served as a document provider. During the test, libraries received documents from UMass via Ariel or via the Web as a PDF document. This form of document delivery was faster than standard service with better quality of delivered articles. The column describes the project and outlines possible future steps. PMID- 14527141 TI - Integration of information literacy into a revised medical school curriculum. AB - The Keck School of Medicine of the University of Southern California's newly revised Year I-II curriculum integrates information literacy components based on the AAMC's medical informatics objectives throughout the two-year continuum. Students are required to participate in various components taught by librarians from the Norris Medical Library including (1) an information skills workshop, (2) a yearlong literature search project, and (3) an evidence-based medicine resources workshop. Critical appraisal of literature also is included in the curriculum to ensure that the students are familiar with the various research study designs. Librarians also provide resources to assist students in researching learning issues related to cases presented in the curriculum. To encourage the faculty mentors to serve as good models of information users, librarians offer brief training sessions on information resources to faculty. PMID- 14527143 TI - Amination and [2,3]-sigmatropic rearrangement of propargylic sulfides using a ketomalonate-derived oxaziridine: synthesis of N-allenylsulfenimides. AB - Amination of propargylic sulfides with a ketomalonate-derived oxaziridine under metal free conditions gives N-Boc-N-allenylsulfenimides via [2,3]-sigmatropic rearrangement. PMID- 14527142 TI - Preparation of 3-acetoacetylaminobenzo[b]furan derivatives with cysteinyl leukotriene receptor 2 antagonistic activity. AB - Novel 3-acetoacetylaminobenzo[b]furan derivatives with a modified triene system at the 3-position were prepared through acylation of the 3-aminobenzo[b]furans with 5-methylisoxazole-4-carboxylic acid chloride followed by basic cleavage of the isoxazole ring and several of these compounds showed moderate cysteinyl leukotriene receptor 2 antagonistic activity. PMID- 14527144 TI - Alkali metal ion complexes of functionalised calixarenes--competition between pendent arm and anion bonding to sodium. AB - Determination of the crystal structure of the acetonitrile inclusate of the complex formed between sodium trifluoromethanesulfonate (triflate, CF3SO3-) and the narrow-rim functionalised calix[4]arene, 5,11,17,23-tetra-tert-butyl-25,27 di(phenylmethoxy)-26,28-di(2'-methoxyethoxy)calix[4]arene, has shown, somewhat unexpectedly, that the diether pendent arms do not chelate the sodium cation, although coordination of all four phenolic oxygen atoms does draw the calixarene into a nearly symmetrical cone form, consistent with conclusions drawn earlier from solution 1H NMR data. Crystals of C64H80O6.NaO3S.CF3.CH3CN obtained from acetonitrile solvent are monoclinic, C2/c, a structure determination at 'low' temperature (153 K) resolving several difficulties encountered in earlier attempts to analyse data acquired at approximately 295 K, and indicative of an interesting temperature dependence of substituent and anion orientations. PMID- 14527145 TI - Efficient synthesis of glycosylated phenazine natural products and analogs with DISAL (methyl 3,5-dinitrosalicylate) glycosyl donors. AB - Inspired by the occurrence and function of phenazines in natural products, new glycosylated analogs were designed and synthesized. DISAL (methyl 3,5 dinitrosalicylate) glycosyl donors were used in an efficient and easily-handled glycosylation protocol compatible with combinatorial chemistry. Benzoylated D glucose, D-galactose and L-quinovose DISAL glycosyl donors were synthesized in high yields and used under mild conditions to glycosylate methyl saphenate and 2 hydroxyphenazine. The glycosides were screened for biological activity and one compound showed inhibitory activity towards topoisomerase II. PMID- 14527146 TI - The Co(I) induced methylmalonyl-succinyl rearrangement in a model for the coenzyme B12 dependent methylmalonyl-CoA mutase. AB - The rearrangement of 2-bromomethyl-2-methylmonothiomalonates to succinyl derivatives was found to take place in quantitative yields in the presence of one molar equivalent of Co(I) generated by the reduction of heptamethyl Co(II)yrinate perchlorate with NaBH4 or electrochemically. The chiral thiomalonate gave racemic succinate. PMID- 14527147 TI - Conversion of 2-deoxy-D-ribose into 2-amino-5-(2-deoxy-beta-D ribofuranosyl)pyridine, 2'-deoxypseudouridine, and other C-(2' deoxyribonucleosides). AB - The synthesis of 2-amino-5-(2-deoxy-beta-D-ribofuranosyl)pyridine 2a, 2-amino-5 (2-deoxy-alpha-D-ribofuranosyl)-pyridine 23, 2-amino-5-(2-deoxy-beta-D ribofuranosyl)-3-methylpyridine 2b, 2-amino-5-(2-deoxy-alpha-D-ribofuranosyl)-3 methylpyridine 29 and 5-(2-deoxy-beta-D-ribofuranosyl)-2,4-dioxopyrimidine [2' deoxypseudouridine] 30a is described. These C-nucleosides are prepared either from 2-deoxy-3,5-O-(1,1,3,3-tetraisopropyldisiloxan-1,3-diyl)-D-ribofuranose 15 or from 2-deoxy-3,5-O-(1,1,3,3-tetraisopropyldisiloxan-1,3-diyl)-D-ribono-1,4 lactone 16, which are themselves prepared from 2-deoxy-D-ribose 13. The sugar derivatives are first allowed to react with the appropriate 5-lithio-pyridine or 5-lithio-pyrimidine derivatives, which are prepared from 5-bromo-2 (dibenzylamino)pyridine 12a, 5-bromo-2-[bis(4-methoxybenzyl)amino]pyridine 12b, 5 bromo-2-dibenzylamino-3-methylpyridine 25 and 5-bromo-2,4-bis(4 methoxybenzyloxy)pyrimidine 33. The products from the reactions between the lithio-derivatives and the lactol 15 are cyclized under Mitsunobu conditions; the products from the reactions between the lithio-derivatives and the lactone 16 are first reduced with L-Selectride before cyclization, also under Mitsunobu conditions. In all cases, the beta-anomers of the protected C-nucleosides are the predominant products. Finally, the separation of the alpha- and beta-anomers and the removal of all of the protecting groups are described. PMID- 14527148 TI - Rapid and flexible synthesis of 1-deoxy-D-xylulose-5-phosphate, the substrate for 1-deoxy-D-xylulose-5-phosphate reductoisomerase. AB - 1-Deoxy-D-xylulose-5-phosphate (DXP) is a key intermediate in the non-mevalonate pathway to terpenoids in bacteria, and it is the substrate for the enzyme 1-deoxy D-xylulose-5-phosphate reductoisomerase (DXP-R). In order to study the mechanism of DXP-R, we required a flexible synthesis of the substrate which would allow the incorporation of isotopic labels, and the variation of the two stereocentres. Thus 1,4-dihydroxypent-2-yne was selectively reduced to give the E-olefin, and selective phosphorylation of the primary alcohol followed by oxidation of the secondary alcohol gave a substrate suitable for dihydroxylation. Dihydroxylation using stoichiometric OsO4 in the presence of chiral ligands gave protected DXP in high ee. Final hydrogenolysis gave DXP in quantitative yield and high purity. DXP R was produced by rapid cloning of the dxr gene from Escherichia coli through controlled expression and ion exchange chromatography. The synthetic DXP was fully active in enzyme assays catalysed by recombinant DXP-R. PMID- 14527150 TI - Fluorescence studies on nyctinasty which suggest the existence of genus-specific receptors for leaf-movement factor. AB - Periodic leaf-movement of legumes is called nyctinasty and has been known since the age of Alexander the Great. We found that nyctinasty is controlled by a periodic change of the internal concentration of leaf-opening and leaf-closing substances in the plant body. Now, we have developed novel fluorescent probes (1) based on the structure of cis-p-coumaroylagmatine (3), which was isolated as a leaf-opening substance of Albizzia juribrissin Durazz. Binding experiments using probe 1 showed that Albizza plants have receptors for a leaf-opening substance in their motor cells. By using probes 1 we then found that genus-specific receptors are involved in nyctinasty. PMID- 14527149 TI - Functional and structural characterization of ovine ornithine transcarbamoylase. AB - Ornithine transcarbamoylase from ovine liver has been purified to homogeneity. Like all anabolic OTCs, the ovine enzyme is a trimer, constituted by identical subunits of 34 kDa. Sequence analysis of the 54 N-terminal residues of ovine OTC shows a high degree of homology with the human enzyme. The optimum pH and the Michaelis constants for the catalytic reaction were determined. The ovine enzyme is the most thermostable one among mammals OTCs, its critical temperature being 6 degrees C higher than those measured for the other enzymes. The enzyme has been crystallised and the structure determined at 3.5 A resolution. Crystals belong to the cubic P4(3)32 space group, with a = b = c = 184.7 A and a solvent content of about 80%. There is no evidence of any ligand in the active site cavity, indicating that the crystals contain an unliganded or T state of the enzyme. The unliganded OTCase enzyme adopts a trimeric structure which, in the crystal, presents a three-fold axis coincident with the crystallographic one. The conformation of each monomer in the trimer is quite similar to that of the liganded human protein, with the exception of a few loops, directly interacting with the substrate(s), which are able to induce a rearrangement of the quaternary organisation of the trimer, that accounts for the cooperative behaviour of the enzyme following the binding of the substrates. PMID- 14527151 TI - An enantioselective synthesis of the C(33)-C(37) fragment of Amphotericin B. AB - An enantioselective synthesis of the C(33)-C(37) tripropionate fragment of Amphotericin B has been developed in only 6 steps. PMID- 14527152 TI - Reductive decomplexation of pi-allyltricarbonyliron lactone complexes using sodium naphthalenide as a route to stereodefined 1,7-diols and 2,3-diene-1,7 diols. AB - Treatment of pi-allyltricarbonyliron lactone complexes, that contain an adjacent leaving group, with lithium naphthalenide causes decomplexation to acyclic dienols in excellent yield and without any stereochemical scrambling of the allylic centre. When an endo complex is employed (E,E)-geometry prevails with good selectivity whereas (Z,E)-geometry dominates in the case of exo complexes. A mechanism consistent with the observed stereo- and regiochemistry is proposed. PMID- 14527153 TI - 1,5-asymmetric induction of chirality using pi-allyltricarbonyliron lactone complexes: highly diastereoselective synthesis of alpha-functionalised carbonyl compounds. AB - Silyl enol ethers derived from ketone functionalised rho-allyltricarbonyliron lactone complexes undergo highly diastereoselective carbon-fluorine and carbon oxygen bond formation reactions with excellent control at the alpha-stereogenic centre. PMID- 14527154 TI - Reappraising the formation of Jaffe's Base: studies of the treatment of imidazolidine-2-thione with mild oxidising agents. AB - Throughout reports detailing the preparation of Jaffe's Base, which date as far back as 1894, its identity and the nature of an intermediate prior to its formation have been constant points of conjecture. This report presents firm evidence that the oxidation of imidazolidine-2-thione, SCN(H)C2H4N(H), with potassium triiodide yields the 'thiol' condensation product 2,2'-bis(4,5-dihydro 1H-imidazolidine)disulfide (NC2H4N(H)CS)2 (1), which under mild conditions undergoes partial self condensation to yield 1-(4,5-dihydro-1H-imidazolidin-2 yl)imidazolidine-2-thione, Jaffe's Base--SCN(CNC2H4N(H))C2H4NH, (2). Crystallisation of 1 from acetone results in the unexpected formation of heterobicyclic 3-methyl-5,6-dihydroimidazolidin[2,1-b]thiazole, 3. The conversion of 1 to 3 has been studied by 1H NMR spectroscopy, which suggests the concurrent formation of 2 as an unreactive by-product. The solid state structures of 2, its HBr salt (2.HBr) and 3.(HI.I2), the latter being the isolated form of 3 under the conditions employed, have been determined using XRD methods. PMID- 14527155 TI - First synthesis of etidronate partial amides starting from PCl3. AB - Methods for the preparation of mixed tetra-amide esters 1 and 2, the partial amide ester 3, and tri- and P,P-diamides 4 and 5 from monophosphorus spieces 12, 8 and 9, respectively, were developed. Compounds 8 and 9 were obtained from phosphorus trichloride via MeOPCl2, which was treated with 2 eq. and 4 eq. of piperidine, followed by water or acetyl chloride, respectively. Tetrasubstituted amide bisphosphonates 1 and 2 were selectively dealkylated with lithium or silyl halide to achieve target compounds 3-5. Piperidine was found to be a good desilylation reagent. Quantum mechanical calculations illustrate why derivative 2 was produced in low yield. The usefulness of compounds 1, 3 and 4 as prodrugs of etidronate was determined in aqueous buffer and human serum. PMID- 14527156 TI - Imidazolylidene carbene ligated palladium catalysis of the Heck reaction in the presence of air. AB - Five 1,3-disubstituted imidazolium salts were synthesized. Their Heck reaction activities were evaluated. A convenient, efficient and high yielding procedure based on these compounds for the arylation of olefins was developed. Heck reactions mediated by these palladium-N-heterocyclic carbene complexes were conducted under air and even in the presence of several common oxidants. PMID- 14527157 TI - Cu(II)-nitroxyl radicals as catalytic galactose oxidase mimics. AB - Results from Hammett correlation studies and primary kinetic isotope effects for the CuCl-TEMPO catalysed aerobic benzyl alcohol oxidations are inconsistent with an oxoammonium based mechanism. We postulate a copper-mediated dehydrogenation mechanism, in which TEMPO regenerates the active Cu(II)-species. This mechanism is analogous to that observed for Galactose Oxidase and mimics thereof. PMID- 14527158 TI - Towards more chemically robust polymer-supported chiral catalysts: alpha,alpha diphenyl-L-prolinol based catalysts for the reduction of prochiral ketones with borane. AB - alpha,alpha-Diphenyl-L-prolinol derivatives with para-bromo substituents in either one or both of the phenyl rings are easily bound to crosslinked polystyrene beads containing phenylboronic acid residues by Suzuki couplings. By using extended reaction periods boronic acid residues that do not take part in the couplings are simply lost by hydrolysis. The polymer-supported (PS) alpha,alpha-diphenyl-L-prolinols were used to catalyse reductions of several prochiral ketones with borane in tetrahydrofuran at 22 degrees C. The expected secondary alcohols were obtained in high chemical yields and ees were generally in the range 79-97%. One PS catalyst was recycled 14 times without loss of stereochemical performance. PMID- 14527159 TI - Molecular screening on a compact disc. AB - A method is described to screen the recognition between small molecule ligands and biomolecules using a conventional compact disc (CD) player. A procedure was developed to attach ligands to the reading face of a CD by activating the terminus of polycarbonate, a common polymer composite within the reading face of a CD. Terminal residues of the polycarbonate surface 1 were phosphorylated by reaction with ethyl-(N,N)-diisopropylamine-buffered dichloro-(N,N) diisopropylaminophosphate to yield surface-bound chlorophosphate 2. Ligands containing a primary alcohol were condensed with 2 providing a polycarbonate capped with phosphodiester linked ligands 3-6. Displays were generated on the surface of a CD by printing tracks of ligands 3-6 on the disc with an inkjet printer. Using this method, discs were created with entire assemblies of ligand molecules distributed into separate blocks. A molecular array was developed by assembling collections of these blocks to correlate with the CDROM-XA formatted data stored within the digital layer of the disc. Regions of the disc containing a given ligand or set of ligands was marked by its spatial position using the tracking and header information. Recognition between surface expressed ligands and biomolecules was screened by an error determination routine. PMID- 14527160 TI - An NMR and X-ray study of the structure of the azo coupling product of 4 dimethylaminopent-3-en-2-one and benzenediazonium-tetrafluoroborate. AB - 4-Dimethylaminopent-3-en-2-one reacts with two molecules of benzenediazonium tetrafluoroborate to give compound 1. The structure of this compound was determined by means of X-ray analysis of its crystal and 1H, 13C and 15N NMR spectra of its solution in CDCl3. The molecule of this compound contains one azo group and one hydrazone group. The substance exists, both in crystal form and in solutions of concentrations above 0.1 mol l(-1), in the form of a dimer, in which the pair of molecules are bound by two hydrogen bonds N-H...N. On diluting the solution, the dimers decompose, the two forms being in an equilibrium that is rapid on the NMR time scale. PMID- 14527161 TI - Perisynaptic barrier of proteoglycans in the mature brain and spinal cord. AB - Cell bodies and their dendrites of motor neurons, motor-related neurons, and certain other subsets of neurons such as GABAergic interneurons in the mature brain and spinal cord possess intensely negatively charged perineuronal or perisynaptic nets of proteoglycans which are linked to the nerve cell surface glycoproteins. These perineuronal nets of proteoglycans are digested by chondroitinase ABC, hyaluronidase, or collagenase, but not by endo-alpha-N acetylgalactosaminidase, which is reactive to the nerve cell surface glycoproteins. Aggrecan, versican, neurocan, and brevican are members of a family of chondroitin sulfate proteoglycans that bind to hyaluronan. Neurocan- or brevican-deficient mice showed a regionally heterogeneous composition of proteoglycans in perineuronal nets. Aggrecan glycoforms contribute to the molecular heterogeneity of the perineuronal nets. Proteoglycans such as phosphacan are included in matrix-associated proteoglycans. The extracellular matrix glycoprotein tenascin-R is accumulated in the perineuronal nets. The perineuronal proteoglycans are produced by associated satellite astrocytes just before weaning, while the nerve cell surface glycoproteins are produced by the associated nerve cells at earlier stages after birth. The perineuronal proteoglycans may entrap the tissue fluid and form a perineuronal gel layer which protects the synapses as a "perisynaptic barrier". Degradation of the perineuronal proteoglycans or perisynaptic barrier by treatment with chondroitinase ABC or hyaluronidase reactivates the neuronal plasticity or promotes the functional recovery of a severed nervous system. Another set of perineuronal nets occurs, which are intensely positively charged and contain guanidino compounds. It is considered that these intensely positively charged nets are intermingled with the intensely negatively charged ones of proteoglycans. PMID- 14527162 TI - Retrograde labeling of mouse spinal descending tracts by a recombinant adenovirus. AB - The present study tested whether a gene-transfer based upon the retrograde axonal transport of the lacZ adenovirus is effective in the spinal descending tracts of the adult mouse. A small volume of a replication-defective recombinant adenovirus encoding E. coli beta-galactosidase was injected into the upper lumbar cord, and, seven days later, the mice were transcardially perfused by a fixative solution. X gal staining of coronal or sagittal sections of the spinal cord and the brain revealed that many sites of origin for rubrospinal, vestibulospinal, and reticulospinal tracts were retrogradely labeled, whereas few of the corticospinal tract neurons were retrogradely labeled. Ependymal cells surrounding the central canal of the spinal cord, which were located far from the injection site, showed a high expression of beta-galactosidase activity. Motoneurons around the injection site were strongly stained by X-gal staining, and their axons in the ventral root were anterogradely labeled. Afferent fibers in the dorsal root were labeled by the transganglionic transport of beta-galactosidase. To examine the efficacy of the uptake and retrograde transport of HRP and adenovirus, we injected a mixed solution of 10% HRP and recombinant adenovirus. The number of HRP-labeled corticospinal neurons overwhelmed the number of X-gal stained ones, while the numbers of HRP-labeled rubrospinal and subcoeruleus-spinal neurons were smaller in comparison with the numbers of beta-galactosidase-positive counterparts. The present study revealed that the origins for the spinal descending tracts except for corticospinal neurons could be efficiently gene transferred by the retrograde infection of a recombinant adenovirus. Such a difference in efficacy of retrograde infection among the spinal descending tracts is practically important when an adenovirus-mediated gene transfer is designed to treat certain neurological diseases affecting the spinal descending tracts. PMID- 14527164 TI - The effects of sex steroids on the formation of gap junctions between folliculo stellate cells; a study in castrated male rats and ovariectomized female rats. AB - We investigated the relationship between gap junction formation and the sex steroids testosterone, progesterone and 17beta-estradiol in the anterior pituitary glands of castrated male rats and ovariectomized female rats. Male and female 30-day-old Wistar-Imamichi strain rats were castrated or ovariectomized, and 30 days later they were subcutaneously injected with the above sex steroids. They were divided into six groups according to the injected materials: sesame oil (control), testosterone, progesterone, 17beta-estradiol, testosterone with 17beta estradiol, and progesterone with 17beta-estradiol. Five rats from each group were sacrificed 1, 2, 3, 4 and 5 days after the injections, and the anterior pituitary glands were prepared for observation by transmission electron microscopy. We quantified the number of follicles and gap junctions and calculated the rate of occurrence of gap junctions as the ratio of the number of gap junctions existing between folliculo-stellate cells per intersected follicle profile in electron photomicrographs. The administration of testosterone to castrated male rats increased the rate of gap junctions between folliculo-stellate cells; however, progesterone and 17beta-estradiol did not affect the formation of gap junctions. The administration of progesterone to ovariectomized female rats increased the rate of gap junctions between folliculo-stellate cells; this progesterone effect was prevented by the simultaneous administration of 17beta-estradiol, which by itself did not affect the rate of gap junctions between folliculo-stellate cells. These observations indicate that the formation of gap junctions within the anterior pituitary gland is regulated differently by sex steroids in castrated male and ovariectomized female rats. PMID- 14527163 TI - Chondromodulin-I expression in rat articular cartilage. AB - The localization and expression of chondromodulin-I (ChM-I), an angiogenesis inhibitor, in the rat articular cartilage during maturation from 2 to 10 weeks of age were examined by immunohistochemistry, Western blot analysis and ribonuclease protection assay, and the results were compared with those in the epiphyseal cartilage. ChM-I was found to be diffusely immunostained in the inter-territorial space of the cartilage matrix from the intermediate to the deep layers at the immature stage. As the articular cartilage matured, the immunoreactivity was localized around the hypertrophic chondrocytes in the deep layer and the immunoreactivity became weak after maturation. In contrast, the ChM-I immunoreactivity was intense in the epiphyseal cartilage at all ages examined. ChM-I was detected by Western blotting as a broad band or occasionally as a cluster of multiple bands (approximately 25 kDa) in both the articular and the epiphyseal cartilage. The intensity of the bands decreased gradually with age in the articular cartilage, but was unchanged in the epiphyseal cartilage at all ages. Ribonuclease protection assay revealed that ChM-I mRNA also decreased gradually with age in the articular cartilage in parallel with the maturation of the articular cartilage, while no decrease in ChM-I mRNA was found in the epiphyseal cartilage. The expression of ChM-I mRNA in the articular cartilage was less than that in the epiphyseal cartilage at all ages. The decrease in amount of ChM-I in the mature articular cartilage suggests that ChM-I plays a more important role in the maintenance of avascularity in the immature articular cartilage than in the mature one. The avascular condition may be preserved by angiogenic inhibitors or mechanisms other than ChM-I in the mature articular cartilage. PMID- 14527165 TI - Pathways for movement of fluid and cells from hepatic sinusoids to the portal lymphatic vessels and subcapsular region in rat livers. AB - It has long been a mystery how fluid and migrating cells in the hepatic sinusoids reach lymphatic vessels in the portal tract. Here we describe previously-unknown channels that connect the space of Disse with the portal tract in the rat liver. Transmission electron microscopy was performed on livers injected with either horseradish peroxidase (HRP) or lipopolysaccharide, and scanning electron microscopy was carried out on livers macerated with KOH. Transmission electron microscopy revealed the presence of channels with collagen fibers traversing the limiting plate. A tracer study showed that HRP was in the channels as well as along inlet venules. Dendritic cells in the hepatic sinusoids or between hepatocytes of the limiting plate were also observed extending their pseudopodia through the channels in the limiting plate to the interstitial space of the portal tract. Scanning electron microscopy further showed that many channels (1 3microm in diameter) penetrated through the limiting plate independently of blood vessels and connected the space of Disse with the interstitial space of the portal tract. In addition, the portal tract possessed prelymphatic vessels that were lined with fibroblast-like cells and frequently contained dendritic cells. The initial segment of the portal lymphatic vessels opened to the interstitial tissue space. These results indicate that fluid and dendritic cells in the hepatic sinusoids probably pass through both the space of Disse and the channels traversing the limiting plate, enter the interstitial space of the portal tracts, and finally move from the prelymphatic vessels to the portal lymphatic vessels. PMID- 14527166 TI - Expression of leptin receptor (Ob-R) isoforms and signal transducers and activators of transcription (STATs) mRNAs in the mouse taste buds. AB - Leptin is a hormone that regulates food intake, energy expenditure and body weight. Our previous studies have demonstrated that the taste organ is a new peripheral target for leptin in mice. Leptin selectively inhibits the responses of taste nerves and receptor cells to sweet substances without affecting responses to sour, salty, and bitter substances. Still, there is no convincing evidence for the existence of leptin receptors (Ob-Rs) in taste receptor cells, especially the functional isoform Ob-Rb. We investigated the expression of 5 different Ob-R isoforms (a-e) and 6 STAT (signal transducers and activators of transcription) members in mouse taste cells. STATs are considered to be involved in the leptin signaling via Ob-Rb. Semiquantitative RT-PCR analysis showed that Ob-Rb was expressed in the taste buds of the fungiform and circumvallate papillae, but not so clearly in the surrounding epithelial tissue. The expression pattern among the three different tissues was similar to that of the taste cell specific G-protein, alpha-gustducin. The other Ob-R isoforms were widely detected in either the taste papillae or the epithelial tissue. Among 6 STAT members, STAT3 showed the highest relative abundance of mRNA in the taste buds. Consistently, in situ hybridization analysis showed that while Ob-Rb and STAT3 signals were detected in some taste bud cells, the signals were not clearly observed in the epithelial tissue cells. In conclusion, the present study provides evidence of the existence of the leptin receptor, Ob-Rb, and STAT3 in the mouse taste bud cells. This finding further confirms the involvement of leptin in the control of taste sensitivities to sweet substances in mice. PMID- 14527167 TI - Fluid and cellular pathways of rat lymph nodes in relation to lymphatic labyrinths and Aquaporin-1 expression. AB - The aim of the present study was to examine the organization of lymph fluid and cellular pathways and distribution of the membrane water channel Aquaporin-1 (AQP 1) in rat lymph nodes. Lymph fluid and cellular pathways within lymph nodes were examined by fluorescent protein tracer/confocal microscopy and by scanning electron microscopy (SEM), While the distribution of AQP-1 was studied immunohistochemically. Tracer studies showed the subcapsular sinuses continued directly at the hilum or via the intermediate sinuses to the medullary sinuses, and lymphatic labyrinths originating with blind-ends in the deep cortex drained into medullary sinuses. Afferent lymph tracers were also observed in node cortex interstitium. By SEM, lymphatic labyrinths appeared densely filled with lymphocytes and had few intraluminal sinus reticular cells, while medullary sinuses possessed well-developed networks of sinus reticular cells. The presence of many lymphocytes wedged in the walls of the lymphatic labyrinth suggested that lymphocytes migrate between the node parenchyma and lymphatic labyrinths. AQP-1 was distributed on the membrane of lymphatic endothelium and reticular cells as well as on both luminal and abluminal cell membranes of high endothelial venules (HEVs). Our SEM findings support the concept that lymphocytes migrate from the node parenchyma into lymphatic labyrinths in the deep cortex. The nodal distribution of AQP-1 plus the presence of a polarized distribution of ion pumps and/or ion channels in the HEV endothelium hypothesized in our discussion could explain the mechanism of the reported lymph-to-plasma fluid flux in lymph nodes and also facilitate the entry of afferent lymph antigens into the node cortex interstitium. PMID- 14527168 TI - Structure of the rat subcutaneous connective tissue in relation to its sliding mechanism. AB - Mammalian skin can extensively slide over most parts of the body. To study the mechanism of this mobility of the skin, the structure of the subcutaneous connective tissue was examined by light microscopy. The subcutaneous connective tissue was observed to be composed of multiple layers of thin collagen sheets containing elastic fibers. These piled-up collagen sheets were loosely interconnected with each other, while the outer and inner sheets were respectively anchored to the dermis and epimysium by elastic fibers. Collagen fibers in each sheet were variable in diameter and oriented in different directions to form a thin, loose meshwork under conditions without mechanical stretching. When a weak shear force was loaded between the skin and the underlying abdominal muscles, each collagen sheet slid considerably, resulting in a stretching of the elastic fibers which anchor these sheets. When a further shear force was loaded, collagen fibers in each sheet seemed to align in a more parallel manner to the direction of the tension. With the reduction or removal of the force, the arrangement of collagen fibers in each sheet was reversed and the collagen sheets returned to their original shapes and positions, probably with the stabilizing effect of elastic fibers. Blood vessels and nerves in the subcutaneous connective tissue ran in tortuous routes in planes parallel to the unloaded skin, which seemed very adaptable for the movement of collagen sheets. These findings indicate that the subcutaneous connective tissue is extensively mobile due to the presence of multilayered collagen sheets which are maintained by elastic fibers. PMID- 14527169 TI - The organization of the lamina muscularis mucosae in the human esophagus. AB - The structural organization of the lamina muscularis mucosae of the human esophagus was studied by light microscopy and scanning electron microscopy (SEM). The organization of the lamina muscularis mucosae varied considerably among the cervical, the thoracic, and the abdominal part of the esophagus. In the cervical part, the lamina muscularis mucosae was not well developed and only islets of the smooth muscle bundles were scattered within the connective tissue. In the thoracic part, the lamina muscularis mucosae consisted of several layers of smooth muscle bundles, individual muscle cells of which ran in a longitudinal direction. In the abdominal esophagus near the cardia, the muscular bundles in the lamina muscularis mucosae ran in various directions forming a reticular configuration. The differences in density and arrangement of the lamina muscularis mucosae are discussed in relation to the swallowing of food and submucosal invasion of esophageal cancer. PMID- 14527171 TI - Tyloxapol attenuates the pathologic effects of endotoxin in rabbits and mortality following cecal ligation and puncture in rats by blockade of endotoxin receptor ligand interactions. AB - We have previously demonstrated that the detergent Tyloxapol is effective in preventing reactions to endotoxin. We studied the effects of Tyloxapol on the morbidity and mortality from endotoxemia in rabbits and on the mortality in rats with sepsis. The effects of Tyloxapol on endotoxin binding and macrophage activation were studied in the macrophage cell line RAW264.7 and CHO cells expressing CD14. Isolated human leukocytes were used to study the effects of Tyloxapol on immune reactions, leukocyte motility, and phagocytosis. Intravenous Tyloxapol (200 mg/kg), given prior to or at the time of endotoxin infusion protected rabbits from developing shock. In rats with peritoneal sepsis, a lipid rich diet and Tyloxapol given at the time of induction of peritonitis protected them from septic death. In vitro, Tyloxapol blocked the binding of endotoxin to murine macrophages and CHO cells expressing CD14, activation of macrophages, and also some antigen-antibody immune reactions (mediated by CD2, CD4, CD22, HLA-DR). Tyloxapol may prevent the reaction to endotoxin by desensitizing endotoxin recognizing receptors. PMID- 14527170 TI - Impairment of leukocyte trafficking in a murine pleuritis model by IL-4 and IL 10. AB - We have characterized leukocyte migration to the pleural cavity in a methylated BSA (mBSA)-induced model of murine delayed-type hypersensitivity and evaluated the ability of IL-4 and IL-10 to modulate this response. Neutrophils, macrophages, T cells, and dendritic cells migrated to the pleural cavity in a time-dependent fashion following direct intrapleural antigen challenge, with neutrophils comprising the majority of exudate leukocytes in the cavity within the first 24 h and the number of mononuclear cells increasing at later times. Real-time quantitative PCR analysis of infiltrating leukocytes revealed a marked elevation of steady-state mRNA levels of IL-1beta and TNFalpha and the chemokines KC, MIP-2, CXCL9, CXCL10, CXCL11, CCL2, CCL3, and CCL4 at 6 h postchallenge, which diminished over time. In contrast, gammaIFN mRNA levels were maximal at 24 h and CCL5 expression was sustained throughout 72 h. ELISA analysis of pleural exudate fluid revealed significant elevations of KC and CCL2 protein levels at 6 h postantigen challenge and a peak increase in gammaIFN protein at 24 h, confirming our mRNA observations. Administration of recombinant murine IL-4 or IL 10 prior to challenge significantly blocked cell trafficking to the pleural cavity as well as peak levels of exudate gammaIFN, with IL-4 being more potent in impairing these responses. IL-4 administration also increased the proportion of naive T cells in the pleural cavity, as judged by CD62L and CD45RB expression. These results indicate that this in vivo model demonstrates a pattern of events associated with Th1-mediated leukocyte trafficking and underscore the potential utility of this in vivo model for evaluating therapeutic inhibitors of leukocyte trafficking. PMID- 14527172 TI - Interaction of B7RP-1 with ICOS negatively regulates antigen presentation by B cells. AB - Stimulation of T cells through the T cell receptor is insufficient for optimal T cell activation. A second activation signal is necessary, being usually provided by the costimulatory molecule CD28. Recently, additional costimulatory pathways have been identified, including inducible costimulator (ICOS) and its ligand B7RP 1. We have examined the role of the B7RP-1/ICOS costimulatory pathway on antigen presentation by B cells, using the I-Ak and I-Ek-positive CH27 B cell line and several different T cell lines. We found that CH27 expressed B7RP-1 and PD-L1 whereas the T cell lines expressed ICOS and PD-1. In the presence of HEL, the T cell hybridomas C10 and 3A9 released IL-2, which is indicative of antigen specific T cell activation by the CH27 cells. Unexpectedly, blocking antibodies for B7RP-1 and ICOS enhanced the IL-2 response in both T cells. As expected, an increase in the production of IL-2 was seen when blocking antibodies for PD-1 were used. Blocking with antibodies for I-Ak, CD28, B7.1, and B7.2 lead to a decrease in IL-2 production. Additionally we tested a Th1 and a Th2 T cell clone. Blockade of B7RP-1/ICOS lead to an increased IFN-gamma response in Th1 cells (A.E7) and an increased IL-4 response in Th2 cells (D10.G4.1). Intracellular staining also showed an increase in cytokine production when the B7RP-1/ICOS pathway was blocked. In conclusion, the B7RP-1/ICOS pathway is negatively regulating T cell activation by B cells and may play a role similar to that of the PD-L1/PD-1 pathway. PMID- 14527173 TI - Murine macrophages cultured with IL-4 acquire a phenotype similar to that of epithelioid cells from granulomatous inflammation. AB - Epithelioid cells (ECs) found in granulomas are thought to derive from mononuclear phagocytes. Although GM-CSF and/or IL-4 are known to promote cell differentiation their role in the development of ECs has never been demonstrated. Here we showed that mouse macrophages treated exclusively with recombinant IL-4 (rIL-4) differentiate into epithelioid-like cells. Macrophages cultivated with rIL-4 presented a fried-egg shape, and ultrastructural studies revealed membrane interdigitations, cytoplasmic vesicles, prominent Golgi complex, and rough endoplasmic reticulum. Compared with controls, rIL-4 treated cells displayed increased expression of MHC class II molecules and of Migration Inhibitory Factor Related Protein-14. Whereas mannose receptor-mediated phagocytosis was increased, Fcgamma-receptor mediated phagocytosis and the production of nitric oxide were decreased in treated cultures. All these features overlap those reported for ECs from granulomatous lesions. In conclusion, treatment of mouse peritoneal macrophages with rIL-4 drives their in vitro differentiation to an epithelioid phenotype and provides a tool to investigate the biology of ECs. PMID- 14527174 TI - Nitric oxide modulates MCP-1 expression in endothelial cells: implications for the pathogenesis of pulmonary granulomatous vasculitis. AB - Monocyte chemoattractant protein-1 (MCP-1) is a pivotal mediator of angiocentric granuloma formation in glucan-induced pulmonary granulomatous vasculitis. Based on the rationale that mononuclear phagocytes retrieved from granulomas are rich sources of nitric oxide (NO) and that the recruitment of mononuclear phagocytes into lesions abates as granuloma formation slows, we tested the hypothesis that MCP-1 gene expression is regulated by a NO-sensitive mechanism. Preexposure of endothelial cell (EC) monolayers to NO donor compounds markedly reduced cytokine induced MCP-1 expression and cytosolic-to-nuclear translocation of nuclear factor kappa B (NF-kappaB), reversed fluctuations in endothelial reduced glutathione (GSH) pools but did not affect cGMP concentrations. The lungs of mice bearing targeted disruptions of the inducible nitric oxide synthase (iNOS) gene exhibited significantly higher concentrations of MCP-1 following glucan infusion than did those of wild-type mice. Cumulatively, these data suggest that NO suppresses MCP 1 expression by blunting the redox changes associated with cytokine-induced EC activation. PMID- 14527175 TI - Differential activation of signal transduction pathways mediating oxidative burst by chicken heterophils in response to stimulation with lipopolysaccharide and lipoteichoic acid. AB - Toll-like receptors (TLRs) have been previously shown to mediate oxidative burst in chicken heterophils. This study was conducted to begin to map the molecular pathways that regulate TLR-mediated oxidative burst. Peripheral blood heterophils from neonatal chicks were isolated and exposed to known inhibitors of signal transduction pathways for either 20 min (genistein, verapamil, or chelerythrine) or 120 min (pertussis toxin) at 39 degrees C. The cells were then stimulated for 30 min at 39 degrees C with Salmonella enteritidis lipopolysaccharide (LPS) or Staphylococcus aureus lipoteichoic acid (LTA). The heterophil oxidative burst was then quantitated by luminol-dependent chemiluminescence (LDCL). Genistein (a tyrosine kinase inhibitor), verapamil (a calcium channel blocker), chelerythrine (a protein kinase C inhibitor), and pertussis toxin (a G-protein inhibitor) significantly reduced LPS-stimulated oxidative burst in chicken heterophils by 34, 50, 63, and 51%, respectively. Although genistein had a statistically significant effect on reducing LPS-stimulated LDCL biologically it seems to play only a minor role within the oxidative burst pathway. Heterophils stimulated with the gram-positive TLR agonist, LTA, activated a different signal transduction pathway since chelerythrine was the only inhibitor that significantly reduced (72%) LTA-stimulated oxidative burst. These findings demonstrate that distinct signal transduction pathways differentially regulate the stimulation of oxidative burst in avian heterophils. Pertussis toxin-sensitive, protein kinase C dependent, Ca(++)-dependent G proteins appear to regulate oxidative burst of avian heterophils stimulated with gram-negative agonist LPS; whereas, a protein kinase C-dependent signal transduction pathway plays the major role activating the oxidative burst of avian heterophils stimulated with gram-positive agonists. The distinct differences in the response of heterophils to these two agonists illustrate the specificity of TLRs to pathogen-associated molecular patterns (PAMP)s. PMID- 14527176 TI - Rhein inhibits interleukin-1 beta-induced activation of MEK/ERK pathway and DNA binding of NF-kappa B and AP-1 in chondrocytes cultured in hypoxia: a potential mechanism for its disease-modifying effect in osteoarthritis. AB - In the present report, we show that bovine articular chondrocytes cultured in low oxygen tension, i.e. in conditions mimicking their hypoxic in vivo environment, respond to IL-1beta (10 ng/mL) by an increased DNA binding activity of NF-kappaB and AP-1 transcription factors. Incubation of the cells with 10(-5) M rhein for 24 h was found to reduce this activity, particularly in the case of AP-1. Mitogen activated kinases (ERK-1 and ERK-2) were activated by exposure of the chondrocytes to 1-h treatment with IL-1beta. This effect was greater in hypoxia (3% O2) than in normoxia (21% O2). Rhein was capable of reducing the IL-1beta stimulated ERK1/ERK2 pathway whatever the tension of oxygen present in the environment. The level of c-jun protein, an element of AP-1 complex, was increased by exposure of the chondrocytes to IL-1beta after 2, 6, and 24 h. Addition of rhein at 10(-5) M for 24 h did not reduce the c-jun protein amount. The mRNA steady-state levels of collagen type II (COL2A1) and aggrecan core protein were found to be significantly increased by a 24-h treatment with 10(-5) M rhein. This stimulating effect was also observed in the presence of IL-1beta, suggesting that the drug could prevent or reduce the IL-1beta-induced inhibition of extracellular matrix synthesis. IL-1-induced collagenase (MMPI) expression was significantly decreased by rhein in the same conditions. In conclusion, rhein can effectively inhibit the IL-1-activated MAPK pathway and the binding of NF-kappaB and AP-1 transcription factors, two key factors involved in the expression of several proinflammatory genes by chondrocytes. In addition, the drug can reduce the procatabolic effect of the cytokine, by reducing the MMPI synthesis, and enhance the synthesis of matrix components, such as type II collagen and aggrecan. These results may explain the antiosteoarthritic properties of rhein and its disease-modifying effects on OA cartilage, in spite of absence of inhibition at prostaglandin level. PMID- 14527177 TI - Complete Freund's adjuvant promotes the increases of IFN-gamma and nitric oxide in suppressing chronic arthritis induced by pristane. AB - The aim of this study was to determine therapeutic effects of complete Freund's adjuvant (CFA) on the progression and relapsing of pristine-induced arthritis (PIA) and investigate the mechanism involved. Chronic relapsing arthritis was induced by pristine in LEW rats. After onset of arthritis, rats were intradermally injected CFA and rats in control group were injected the same volume of PBS. Arthritis was monitored visually, and joint pathology was examined histologically. Cytokine mRNA expression in inguinal lymph nodes was assessed by RT-PCR. The levels of nitric oxide (NO) in serum were measured by colorimetric assay. The results showed that CFA significantly suppressed the progression and relapsing of PIA. Relapsing rate of PIA in CFA-treated group was 12.5% and it was 85.7% in PBS-control group (P < 0.005). CFA markedly inhibited the infiltration of inflammatory cells and cartilage damage in the joints of CFA-treated rats and promoted the increases of IFN-y mRNA and NO levels. The present study provided an implication that adjuvant therapy may be a new strategy for the treatments of rheumatoid arthritis (RA) and other chronic inflammatory diseases. PMID- 14527178 TI - Serum ferritin increases in hemorrhaged rats that develop acute lung injury: effect of an iron-deficient diet. AB - For unknown reasons, serum ferritin levels increase in patients at risk for and with acute lung injury (ALI). To improve understanding of the relationship between serum ferritin alterations and the development of ALI, we investigated the effect of iron deficiency on the serum ferritin response of rats subjected to hemorrhage. We found that rats fed an iron-deficient diet for 6 weeks had decreased hemoglobin, hematocrit, liver total iron, liver total iron-binding capacity, and liver ferritin concentrations but the same serum ferritin concentrations as rats fed a control diet. Following hemorrhage, serum ferritin concentrations increased rapidly and progressively in rats fed a control diet. Along with increases in serum ferritin concentrations, control diet rats subjected to hemorrhage also had increased lung lavage leukocyte numbers, lung myeloperoxidase activities (lung inflammation), and lung lavage protein concentrations (lung leak) compared to control diet fed rats subjected to sham treatment. By comparison, the serum ferritin concentrations, lung inflammation, and lung leak of hemorrhaged rats fed an iron-deficient diet were decreased compared to hemorrhaged rats fed a control diet. These findings indicate that serum ferritin concentrations increase and acute lung injury develops following hemorrhage in rats fed a control, but not an iron-deficient, diet. A relatively brief exposure to an iron-deficient diet reduces hemorrhage-induced ALI. PMID- 14527179 TI - Some antiepileptic compounds impair learning by rats in a Morris water maze. AB - In the present experiments, we investigated the effects of several commonly employed antiepileptic drugs on the performance of adult rats in a Morris water maze task. We found that phenytoin treatment produced the most deleterious performance impairments across all days of training, and that these performance deficits are not likely due to any general sensorimotor impairments. Carbamazepine had milder, but detectable negative effects, as carbamazepine treated animals exhibited initial acquisition deficits, but rapidly achieved escape levels comparable to controls. In marked contrast, valproate and ethosuximide had no detectable effects on learning in the water maze. These results parallel previous findings in rats treated with these compounds and tested in an instrumental learning task, and are in general agreement with the human clinical literature. To the extent that one might wish to minimize learning deficits associated with maintenance on antiepileptic drugs, phenytoin is definitely not the treatment of choice, while valproate or ethosuximide are apparently much less disruptive. PMID- 14527180 TI - Nicotine vs. ethanol discrimination: extinction and spontaneous recovery of responding. AB - Studies regarding extinction and spontaneous recovery of the discriminative stimulus effects of drugs are limited. Eight rats were initially trained to discriminate nicotine (0.4 mg/kg) vs. ethanol (800 mg/kg). For four rats, itraperitaneal (IP) administrations of nicotine fifteen minutes prior to fifteen minute training sessions served as a discriminative stimulus (SD) for predicting food-reinforced lever pressing (VI-1 min). On other sessions ethanol functioned in predicting nonreinforcement (SA). The stimulus roles of the drugs were counterbalanced for the remaining four rats. SA and SD sessions alternated quasi randomly with two daily sessions at 1000 and 1400 hours. Discriminative control was not disrupted following ten extinction sessions under a non-drug/saline condition, but was disrupted following extinction sessions under the original training drugs. Instances of spontaneous recovery (SR) occurred throughout extinction under the drug condtions. There was no evidence for SR two weeks following extinction, but partial recovery four weeks following the final extinction phase. Contextual status (context renewal) had neither a restorative or disruptive impact on extinguished or discriminated responding, respectively. These results support and extend the limited number of other studies by demonstrating extinction and spontaneous recovery of responding discriminated by two distinct drugs. Some theoretical interpretations regarding history effects and training in the context of drug discrimination are entertained. PMID- 14527181 TI - Nicotine produces a within-subject enhancement of contextual fear conditioning in C57BL/6 mice independent of sex. AB - Nicotine enhances learning including contextual fear conditioning. The present study extends previous work on nicotine and conditioned fear to examine the nature of nicotine's enhancement of contextual fear conditioning and sex differences in contextual fear conditioning in C57BL/6 mice using a within subjects design. Mice were trained by pairing of an auditory stimulus of 80 dB, 6 cps train of broad-band clicks conditioned stimulus (CS) with a 2 sec., 0.35 mA shock unconditioned stimulus (US). Twenty-four hours later mice were tested for freezing in the original context, and one hour later mice were retested in the same context. A 0.5 mg/kg dose of nicotine was given either for three conditions: (1) before training, testing, and retesting; (2) before training and retesting; and (3) before retesting only. The use of a within-subjects design allowed for testing if nicotine would produce state-dependent deficits in contextual fear conditioning. Nicotine did enhance contextual fear conditioning in the groups that received nicotine for both training and testing. Nicotine, however, did not alter freezing when given on training but not testing or testing but not training. No sex differences, however, existed for conditioning or for nicotine's effects on conditioning. These results suggest that nicotine enhanced acquisition and retrieval processes but did not produce state-dependent deficits when administered just for training or just for testing. PMID- 14527183 TI - The S-R information stream: where's the filter? AB - Pavlovian conditioning procedures, in which events such as tastes, lights, and sounds become predictors of food, water, and shocks, have been used for studying the role of the information filter in the selection of conditioned responses. Different models posit the filter at different locations in the S-R stream, but most models suggest either a pre-encoding filter, in which much information is discarded at an early stage of processing, or a post-encoding filter, in which the information is stored but not subsequently expressed in behavior. A selective review of the literature on cue-competition effects reveals a plethora of phenomena that support a post-encoding, but not a pre-encoding filter in Pavlovian processes. PMID- 14527184 TI - Perspectives on military medicine. PMID- 14527182 TI - Cannabinoid agonists and antagonists modulate lithium-induced conditioned gaping in rats. AB - Considerable evidence indicates that conditioned gaping in rats reflects nausea in this species that does not vomit. A series of experiments evaluated the potential of psychoactive cannabinoid agonists, delta-9-THC and HU-210, and non psychoactive cannabinoids, Cannabidiol (CBD) and its dimethylheptyl homolog (CBD dmh), to interfere with the establishment and the expression of conditioned gaping in rats. All agents attenuated both the establishment and the expression of conditioned gaping. Furthermore, the CB1 antagonist, SR-141716, reversed the suppressive effect of HU-210 on conditioned gaping. Finally, SR-141716 potentiated lithium-induced conditioned gaping, suggesting that the endogenous cannabinoid system plays a role in the control of nausea. PMID- 14527185 TI - Anthrax: lessons learned from the U.S. Capitol experience. PMID- 14527186 TI - Surgical simulation: a clinical perspective. PMID- 14527187 TI - Assessment and outcomes in medical education. PMID- 14527188 TI - Opportunities for certification in military medicine. PMID- 14527189 TI - The future of medicine in the U.S. Marine Corps. PMID- 14527190 TI - The United States Army Medical Department Center and School: supporting soldiers into the 21st century. PMID- 14527191 TI - The future of medicine in the United States Air Force. PMID- 14527192 TI - Military unique curriculum: identifying and prioritizing content. AB - We have identified and prioritized a series of objectives that warrant inclusion in the continuum of military medical education. Although participants in the 16th Conference on Military Medicine also discussed whether each objective should be taught at the medical student, resident, or staff physician level, to a large extent this distinction is not helpful, since many, if not most, of these topic areas would likely require incorporation at each of these three levels to achieve the desired level of competence in staff physicians. Incorporation of new curricular elements poses a significant challenge, since it is already difficult to fit the existing curriculum into the available time. It is not reasonable to consider increasing the number of lecture hours. Therefore, it is probable that some elements of the existing curriculum will need to be pared down or eliminated to incorporate new material. In the past, when new material has been added to the existing curriculum, such as when the pathogenesis of human immunodeficiency virus was added, it has generally been done at the individual teacher or at most departmental level. Although this approach has the advantage of having a subject matter expert decide how best to insert new material within the fabric of the existing curriculum, there are a couple of problems with widespread use of this approach. First, some of these new objectives may not fit clearly within an existing course curriculum or department's educational mission. Second, such an approach may not provide the degree of coordination that is necessary to ensure that a new curricular item is adequately covered in all respects, and it may result in unnecessary overlap in instruction when different professors incorporate similar elements. Therefore, the prioritization of newer curricular items, as has been done during this conference, may serve as a useful guide in this process. However, a corresponding effort is needed to identify and prioritize existing curricular components. Decisions need to be made regarding what, within the existing curriculum, should be cut out or reduced to make room for some or all of these new objectives. Finally, a decision needs to be made regarding which of these identified objectives should be inserted and when and within which courses they should be taught. These are important decisions that should not be left to the whims of chance. As the responsible recipients of Uniformed Services University and Health Professions Scholarship Program medical school graduates, the services' Surgeons General need to take an active role in helping to make these decisions. The outcome of these decisions will have a direct impact on the capabilities that these future junior medical officers will bring to the patients they will care for and to the line commanders that they will support. PMID- 14527193 TI - Optimal methods of learning for military medical education. PMID- 14527194 TI - Measuring outcomes for military medical education. PMID- 14527195 TI - Certification in military medicine. PMID- 14527196 TI - A study of Y-chromosome microsatellite variation in sub-Saharan Africa: a comparison between F(ST) and R(ST) genetic distances. AB - Seven Y-chromosome microsatellite loci (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, and DYS393) were analyzed in three populations from sub-Saharan Africa: the Bamileke and Ewondo populations from Cameroon and the Hutu from Rwanda. Complete typing was obtained for 112 individuals, and a total of 53 different haplotypes was observed. The single-locus gene diversity, averaged across populations, ranges from 0.100 for the DYS392 locus to 0.610 for the DYS389I locus. The haplotype diversity ranges from 0.832 (Ewondo) to 0.965 (Hutu), with an intermediate value of 0.918 in the Bamileke. The diversity among Bamileke, Ewondo, Hutu, and other sub-Saharan populations selected from the literature was analyzed using both a classical (F(ST)) and a stepwise-based (R(ST)) genetic distance method. The pattern of interpopulational diversity based on F(ST) was congruent with anthropological knowledge, while that based on R(ST) revealed unexpected and unconvincing population affinities. From a practical point of view, our study indicates that Y-chromosome microsatellite data may provide useful information for analyses of interpopulational diversity among sub-Saharan populations if an adequate number of loci and individuals along with an appropriate genetic distance method are used. On a theoretical ground, we propose that the lesser performance of R(ST) compared to F(ST) could be explained by the important role played by genetic drift in shaping the relationships among examined populations. PMID- 14527197 TI - Genetic and linguistic affinities between human populations in Eurasia and West Africa. AB - This study examines the relationship between genetic distance and linguistic affiliation for five regional sets of populations from Eurasia and West Africa. Human genetic and linguistic diversity have been proposed to be generally correlated, either through a direct link, whereby linguistic and genetic affiliations reflect the same past population processes, or an indirect one, where the evolution of the two types of diversity is independent but conditioned by the same geographical factors. By controlling for proximity, indirect correlations due to common geography are eliminated, and any residual relationships found are likely to reflect common linguistic-genetic processes. Clear relationships between genetic distances and linguistic relatedness are detectable in Europe and East and Central Asia, but not in the Middle East, Southeast Asia, or West Africa. We suggest that linguistic and genetic affiliations will only be correlated under specific conditions, such as where there have been large-scale demic diffusions in the last few thousand years, and relative sedentism in the subsequent period. PMID- 14527198 TI - Epidemic cycles in agricultural populations: a cross-cultural study. AB - A cross-cultural analysis of mortality patterns is of interest to biological anthropologists and genetic epidemiologists. In this paper, we examine four agricultural populations from Costa Rica, Hungary, and the United States in order to determine if they suffered from a cyclical distribution of epidemics. When possible, we look at the mortality time series of adults and children separately. Of the 2 series, only 2 show significant epidemic cycles. Both are in the Hungarian groups and both affect subadults. Otherwise, the Costa Rica, U.S., and adult series of the Hungarian groups do not show any periodicity of mortality peaks. Our results indicate that epidemic cycles are not as ubiquitous in small agricultural groups as the literature would suggest. PMID- 14527199 TI - Hypersensitive PCR, ancient human mtDNA, and contamination. AB - When highly efficient polymerase was used with high cycle numbers (50-60), strong amplifications were observed, but negative controls were also unexpectedly amplified in a study of ancient human mtDNA from 2000-year-old skeletons. The results of a series of tests revealed that the hypersensitive polymerase chain reaction (PCR) generated by higher cycles and the presence of contaminant DNA (though at extremely low levels) should be responsible for the amplification of negative controls. We suggest that PCR sensitivity be optimized to take advantage of highly efficient polymerase and at the same time prevent "background DNA" from becoming "contaminant DNA" and obscuring the analysis of authentic ancient DNA. We propose the use of multiple positive controls when amplifying ancient human mtDNA samples to indicate the sensitivity of individual PCR amplifications and to monitor the contamination levels of modern human DNA. This study provides some suggestions as to how to amplify and analyze ancient human mtDNA when unavoidable and extremely tiny amounts of modern human DNA exist. PMID- 14527200 TI - New data on the world distribution of paraoxonase (PON1 Gln 192 --> Arg) gene frequencies. AB - The physiological role of human paraoxonase (PON), a serum enzyme that hydrolyzes organophosphate insecticides and nerve agents, is not clear. Of the three genes in the paraoxonase gene family, PON1 shows a polymorphism, Gln 192 --> Arg, governed by two common alleles named *Q and *R. These determine two different isoforms associated, respectively, with lower and higher activity towards paraoxon, a toxic metabolic product of the insecticide parathion. The *R allele has often been found associated with an increased risk of coronary heart disease. As human populations tend towards greater exposure to environmental changes, including changes in dietary habits and contact with insecticides or other toxic substances, health risks will change as well. In studying the prevention of these newly emerging risks, it could be important to know the distribution of the two alleles in the various world populations. In this paper we report on the genotype and allele frequencies of this polymorphism in different populations, most of which have never been examined for this polymorphism. Samples were taken from mainland Italy, Sardinia, Ethiopia, Benin, and Ecuador. The *R allele frequencies for the samples were: 0.313, 0.248, 0.408, 0.612, and 0.789, respectively. The data show a large variability in allele frequencies, and, in particular, that PON1 allele distribution depends on membership to different geographic populations. PMID- 14527201 TI - HLA genes in the Chuvashian population from European Russia: admixture of Central European and Mediterranean populations. AB - HLA alleles have been determined for the first time in individuals from the Chuvashian population by DNA typing and sequencing. HLA-A, -B, -DR, and -DQ allele frequencies and extended haplotypes have also been determined, and the results compared to those for Central Europeans, Siberians and other Asians, Caucasians, Middle Easterners, and Mediterranean peoples. Genetic distances, neighbor-joining dendrograms, and correspondence analysis have been performed. Present-day Chuvash speak an Altaic-Turkic language and are genetically related to Caucasians (Georgians), Mediterraneans, and Middle Easterners, and not only to Central or Northern Europeans; Chuvash contain little indications of Central Asian-Altaic gene flow. Thus, present-day Chuvash who speak an Altaic-Turkic language are probably more closely related to ancient Mesopotamian-Hittites and northern European populations than to central Asia-Altaic people. PMID- 14527202 TI - CFTR molecular analysis reveals infrequent allele frequencies in nine cystic fibrosis patients from Sao Paulo State, Brazil. AB - Although cystic fibrosis (CF) is the most common autosomal recessive disorder in whites, it is thought to be relatively rare or, alternatively, underdiagnosed in Latin America. In Brazil, different groups have shown a DF508 mutation frequency ranging from 30.7% to 50.8%. Such variation may be explained by the ethnic differences observed in this country, which is genetically very heterogeneous. We describe the molecular analysis for 32 mutations of the CFTR gene in nine unrelated patients with cystic fibrosis from Sao Paulo State, Brazil. The main observation of this study was the absence of 30 out of the 32 mutations in 12 alleles among these patients. Except for mutations DF508 and N1303K, no other mutation could be detected in any of the studied patients. In one of two alleles, a DF508 mutation was detected in four patients (22% of the total sample) and an N1303K mutation was detected in two patients (11% of the total sample). One patient was a compound heterozygote for DF508/N1303K. Although the sample studied here was small, it may be possible that our patients have infrequent alleles once these can occur at higher frequencies in selected populations and also show relevant regional differences. Additional investigations in a larger sample are currently in progress in our laboratory to confirm our results, and further studies are still needed to determine the frequencies of CF gene mutations in different regions and ethnic groups in the Brazilian population. PMID- 14527203 TI - Association of vitamin D receptor polymorphisms with osteoporosis in mexican postmenopausal women. AB - It has been reported that Vitamin D receptor polymorphisms are associated with osteoporosis, particularly those demonstrated by the BsmI and FokI restriction enzymes. Herein we report the results of a case-control study performed in postmenopausal Mexican women. We studied 65 osteoporotic women (< or = -2.5 SD bone mineral density [BMD] of young normal females) and 57 controls (over 90% > or = -1.5 SD BMD of young normal females. Restriction enzymes BsmI and FokI were used to identify polymorphisms. Odds ratios and their 95% confidence intervals were calculated, and analysis was performed controlling for age as a covariate. The BsmI genotypes revealed a higher frequency of the bb genotype in cases than in controls, contradicting much of the literature that suggests this genotype protects females against osteoporosis. Regarding the FokI genotypes, we were unable to confirm that the FF genotype has a protective effect against osteoporosis. The inconsistencies found in the literature and the results obtained in the present work suggest to us that other genetic and nongenetic factors are involved in the occurrence of osteoporosis, confounding the results of the possible association of osteoporosis and VDR polymorphisms. PMID- 14527204 TI - Y-chromosome DNA haplotype XI in Eastern Europe. AB - Haplotype XI frequencies at the Y-chromosome-specific DNA polymorphism (p49-TaqI) were reported in 639 males originating from 13 different geographic locations in Eastern Europe, where haplotype XI represents the major haplotype. The highest frequencies were obtained from Ukraine (44%), Russia (43.9%), and Hungary (40.7%). Percentages of haplotype XI geographic distribution show a gradient of decreasing frequency from these areas of higher percentages toward southeastern and more western countries in Europe. PMID- 14527205 TI - Response to Brimacombe et al., birth order and its association with the onset of chronic fatigue syndrome. PMID- 14527206 TI - Spiers memorial lecture. The quantum chemistry of d- and f-element complexes: from an approximate existence to functional happiness. AB - The field of modern quantum inorganic chemistry is just over 50 years old, dating back to 1951, when quantitative LCAO molecular orbital theory was developed and ferrocene was discovered. This Lecture provides a survey of the development of the field through about 1980, which has led to its current state. The application of modern quantum chemical techniques are illustrated via two disparate examples from the authors' research group. First, the recent discovery of uranium-noble gas bonds is discussed including the synergy between the theoretical and experimental investigations of this phenomenon. New theoretical results using coupled-cluster [CCSD(T)] methodology is contrasted to the original scalar relativistic density functional theory results. Second, new applications of time dependent density functional theory to the rich photochemistry of a dinuclear organometallic complex, (eta5-C5H5)2Fe2(mu-CO)2(CO)2, are discussed. PMID- 14527207 TI - Some structural and electronic properties of MX3 (M = Ln, Sc, Y, Ti+, Zr+, Hf+; X = H, Me, Hal, NH2) from DFT calculations. AB - The geometry of trivalent homoleptic d0 MX3 (X = H, Me, Hal, NH2) complexes for the entire lanthanide family, neutral group 3 and cationic group 4 metal center complexes have been studied with DFT(B3PW91) calculations. The geometrical parameters are in good agreement with the available experimental data. The degree of pyramidalization of the metal is discussed. The hydride and alkyl complexes are strongly pyramidal. In the case of the halide, a pyramidal structure is preferred for fluoride and the systems become increasingly planar with heavier halides. The geometrical trends with X are similar for group 3, group 4 and lanthanide complexes. However group 3 complexes are almost planar, group 4 strongly pyramidal and lanthanide intermediate. The lanthanide contraction is reproduced. A natural bond orbital (NBO) charge analysis is used to rationalize the results. This highlights the similarities and differences in the M-X bonding in the three families of complexes. In all cases, the pyramidalization is related to the participation of the valence d orbitals in the M-X bonds but the M-X bond is mostly ionic in lanthanide and considerably more covalent for the d transition metals. The hydride and alkyl complexes, which have more covalent character than the halide complexes are more pyramidal. In the case of the halide complexes, the fluoride complexes, in which there is the least population of the M d orbitals, are found to be more pyramidal because the increasing covalency with heavier halide stabilizes the planar structure through dpi-ppi interaction. The electronic metal d-p transition of the free ion is shown to be a good indicator of the pyramidalization at M although these values should only be used qualitatively. The strong ionic character of the Ln-X bond gives a rationale for the more important elongation of the beta Si-C bond in La(CH(SiMe3)2)3 than in La(N(SiMe3)2)3. The elongation is shown to be in part due to the negative hyperconjugation of the lone pair used for the Ln-ligand bonds in the beta bonds. PMID- 14527208 TI - On the nature of the short Pt-Tl bonds in model compounds [H5Pt-TlHn]n-. AB - RHF, DFT and MP2 calculations are reported for the compounds [H5Pt-TlHn]n-, n = 0 2. These serve as analogues for the experimentally known [(NC)5Pt-Tl(CN)n](n-) species. The very short bond between platinum and thallium is discussed. PMID- 14527209 TI - The ground state and electronic spectrum of CUO: a mystery. AB - Results are presented from a theoretical study of the lower electronic states of the CUO molecule. Multiconfigurational wave functions have been used with dynamic correlation added using second order perturbation theory. Extended basis sets have been used, which for uranium were contracted including scalar relativistic effects. Spin orbit interaction has been included using the state-interaction approach. The results predict that the ground state of linear CUO is phi2 with the closed shell sigma(+)0 state 0.5 eV higher in energy. This is in agreement with matrix isolation spectroscopy, which predicts phi2 as the ground state when the matrix contains noble gas atoms heavier than Ne. In an Ne matrix, the experiments indicate, however, that CUO is in the sigma(+)0 state. The change of ground state due to the change of the matrix surrounding CUO cannot be explained by the results obtained in this work and remains a mystery. PMID- 14527210 TI - On the reaction of a uranium atom with a nitrogen molecule: a theoretical attempt. AB - An attempt has been made to study the reaction between a uranium atom and a nitrogen molecule theoretically using multiconfigurational wave functions. The C2v part of the reaction surface has been computed for several electronic states of various spin mulltiplicities. The system proceeds from a neutral uranium atom in its (5f)3(6d)(7s)2, 5L ground state to the linear molecule NUN, which has a 1sigma(+)g ground state and uranium in a formal U(VI) oxidation state. The effect of spin-orbit coupling has been estimated at crucial points along the reaction. These preliminary results shows that the system proceeds from a quintet state for U + N2, via a triplet transition state to the final closed shell molecule. An eventual energy barrier for the insertion reaction is caused by the spin orbit coupling energy. PMID- 14527211 TI - Theoretical studies of organometallic complexes of uranium involving nitrogen ligands using density functional approaches. AB - Density functional calculations are used to investigate the structure and bonding in several unusual cyclopentadienyl complexes of uranium with nitrogen-containing ligands. The U(VI) imido complex Cp2U(NPh)2 and the U(IV) amido complex Cp2(NHPh)2 are examined and the important orbitals involved in the U-N bonds are analyzed. The recently synthesized 22-electron U(IV) hydrazonato complex U(IV) Cp*2U(Me-N-N=CR2)2 is explored from the standpoint of an expanded valence shell, and the differences between the structures and thermochemistries of U(IV) and Zr(IV) complexes are probed. PMID- 14527212 TI - A theoretical study of the insertion barrier of MAO methylaluminoxane)-activated, Cp2ZrMe2-catalyzed ethylene polymerization: further evidence for the structural assignment of active and dormant species. AB - Density functional theory has been used to calculate the ethylene insertion barrier into the Zr-C bonds of the [Cp2ZrMe](+)[AlMe3Me(A-MAO)]- and [Cp2ZrMe](+)[Me(A-MAO)]- ion pairs where (AlOMe)6 was used to model the active forms of MAO (A-MAO). The results support the proposal that the former is the active and the latter the dormant species in MAO-activated olefin polymerization. For the first insertion, with the active species the trans approach has a lower barrier to insertion than the cis approach in solution, due to the larger ion pair separation. It is likely that the separated ion pairs recombine between subsequent insertions. For the second insertion, we have considered frontside, backside and combined insertion mechanisms showing that the total barrier to insertion is approximately the same in all cases ranging from 13.8 kcal mol(-1) 14.8 kcal mol(-1). For the frontside and combined insertion this barrier is a result of the rotation of the propyl chain: for the backside mechanism it is stems from the ethylene insertion barrier. Comparison of the results for the ion pair and for the naked cation show that in the former case olefin complexation is an endothermic process whereas for the latter it is exothermic. In general, the large ion pair separation prevents the orientation of the anion from exerting much influence on the geometry or energy of the cation. However, when structures with alpha-agostic interactions are considered the ion pair separation is small enough so that the orientation has some influence on the reaction mechanism. For the second insertion, dissociated transition states were found to have lower barriers than associated ones. PMID- 14527213 TI - Theoretical studies on C-heteroatom bond formation via reductive elimination from group 10 M(PH3)2(CH3)(X) species (X = CH3, NH2, OH, SH) and the determination of metal-X bond strengths using density functional theory. AB - Density functional calculations have been used to investigate C-C, C-N and C-O bond forming reactions via reductive elimination from Group 10 cis M(PH3)2(CH3)(X) species (X = C-I3, NH2, OH). Both direct reaction from the four coordinate species and a three-coordinate mechanism involving initial PH3 loss have been considered. For the four-coordinate pathway the ease of reductive elimination to give M(PH3)2 and CH3-X follows the trend M = Pd < Pt < Ni. The reaction of the cis-M(PH3)2(CH3)(NH2) species is promoted by the formation of methylamine adducts. Non-planar transition states are located and the C heteroatom bond forming processes are characterised by migration of CH3 onto the cis-heteroatom ligand. For a given ligand, X, activation energies follow the trend M = Ni < Pd < Pt. Formation of the three-coordinate M(PH3)(CH3)(X) species is promoted by a labilisation of the cis-PH3 ligand in the four-coordinate reactants when X = NH2 or OH. For the three-coordinate pathway the energy change for reductive elimination to give M(PH3) and CH3-X again follows the trend M = Pd < Pt < Ni and in all cases the initial product is an M(PH3)(XCH3) adduct. The three-coordinate transition states again involve migration of the CH3 ligand onto the cis-X ligand and for X = NH2 or OH activation energies follow the trend Ni > Pd < Pt. For a given metal activation energies in both the four- and three coordinate pathways increase along the series CH3 < NH2 < OH. These trends in activation energy can be rationalised in terms of the strength of M-CH3/M-X bonding as long as the extent of geometrical distortion required to obtain the transition state geometry is taken into account. Further calculations on cis Pd(PH3)2(CH3)(SH) suggest that the more common experimental observation of C(sp3) S compared to C(sp3)-O reductive elimination arises from the greater kinetic accessibility of the former process rather than an intrinsic thermodynamic preference for C-S bond formation. By comparison, the calculations indicate that C(sp3)-N reductive elimination should be feasible from Ni and Pd systems. DF calculations are shown to reproduce the relative homolytic bond strengths determined experimentally for Pt-X bonds. In the cis-M(PH3)2(CH3)(X) systems the M-CH3 homolytic bond strength increases down the group while for M-NH2 and M-OH bonds the trend is M = N approximately equal to Pd < Pt. M-NH2 and M-OH are considerably stronger than M-CH3 bonds and the presence of a heteroatom ligand serves to weaken M-CH3 bonds even further. PMID- 14527214 TI - Modelling spin-forbidden reactions: recombination of carbon monoxide with iron tetracarbonyl. AB - New density functional theory and ab initio computations on the [Fe(CO)5] system are reported. Careful exploration of basis set and correlation effects leads to "best" values for the difference in energy deltaE(1,3) between ground state 3[Fe(CO)4] and the singlet excited state of ca. 8 kcal mol(-1), and for the bond dissociation energy BDE(3) of [Fe(CO)5] with respect to ground state fragments 3[Fe(CO)4] + CO of ca. 40 kcal mol(-1). A modified form of the B3PW91 functional is used to explore the potential energy surface for the spin-forbidden recombination reaction of CO with 3[Fe(CO)4]. A Cs-symmetric minimum energy crossing point (MECP) between the reactant (triplet) and product (singlet) potential energy surfaces is found, lying 0.43 kcal mol(-1) above the reactants. The rate coefficient for recombination is computed using a non-adiabatic form of transition state theory, in which the MECP is treated as the critical point in the reaction. Semi-quantitative agreement with experiment is obtained: the predicted rate coefficient, 8.8 x 10(-15) cm3 molecule(-1) s(-1), is only six times smaller than the experimental rate. This is the first computation from first principles of a rate coefficient for a spin-forbidden reaction of a transition metal compound. PMID- 14527215 TI - Modelling inorganic solids and their interfaces: a combined approach of atomistic and electronic structure simulation techniques. AB - We are seeking to combine the reliability of the structures and energies obtained from quantum mechanical methods with the insights given by larger scale simulations, which are better able to search configurational space. We will discuss our recent work using quantum mechanical methods, based on DFT, which have been applied to the study of a number of solids. Al2O3, CeO2, MnO2 and CaCO3, and compare these with results using atomistic simulation where the forces between atoms are modelled using interatomic potentials. The results show that such quantum methods can be used successfully to screen the different potential models and where necessary, provide sufficient data to allow us to re-consider the potential models. In addition, we show examples where the quantum based methods can give further insights into the reactivity, particularly of surfaces. However, it still remains computationally expensive to search all possible configurations and by using the atomistic simulations to search through different configurations we can identify new structures which can be verified with the quantum based simulations. PMID- 14527216 TI - Multipoles and interaction potentials in ionic materials from planewave-DFT calculations. AB - Oxide potentials which transfer well between different materials have to account explicitly for many-body contributions to the interaction potentials between the ions. These include dipole and quadrupole polarization effects and the compression and deformation of an oxide ion by its immediate coordination environment. Such complex potentials necessarily involve many parameters. We examine how the results of ab initio electronic structure calculations, based upon planewave DFT methods, on general configurations of ions derived from simulations at finite temperature, may be used to parameterize an "aspherical ion method" (AIM) potential (A. J. Rowley, P. Jemmer, M. Wilson and P. A. Madden, J Chem. Phys., 1998, 108, 10209). Dipoles and quadrupoles on the individual ions are obtained via a transformation of the Kohn Sham orbitals to localized orbitals on each ion, which enables a distorted charge density for each ion to be obtained. The dipoles and quadrupoles appearing in polarization parts of the AIM potential are fit to those obtained from the ab initio ionic charge densities obtained in this way. The remaining parts of the potential, describing short range repulsive interactions between ions with compressed and deformed charge densities, are fit to the ab initio forces and the stress tensor. By using a sufficiently large and varied set of configurations on which to carry out this optimization, an excellent transferable potential is obtained. PMID- 14527217 TI - Understanding the interface between oxides and metals. AB - Computational methods based on DFT are applied to modelling structure and bonding in oxide supported metals. We examined three systems of contemporary interest: Cu/ZnO, Pd,Pt/ZrO2 and bimetallic transition metal clusters on silica supports. PMID- 14527218 TI - A theoretical study of isomerism in doped aluminum MAl12 and MAl12X12 clusters with 40 and 50 valence electrons. AB - Systematic density functional B3LYP calculations with the 6-31G* and 6-311+G* basis sets have been employed in order to investigate the structure, vibrational frequencies, relative energies, vertical ionization potentials, and magnetic shielding constants of endohedral and exohedral isomers in two related families of doped aluminum MAl12 and alane MAl12H12 clusters with 40 and 50 valence electrons, respectively. Isomerization barriers have been also determined. Trends in these properties with changing heteroatom M in various series have been followed. The similarities and differences between the aluminides and alanes as well as between the alanes MAl12H12 and related boranes MB12H12 and gallanes MGa12H12 have been scrutinized. PMID- 14527219 TI - Towards an understanding of the bonding in polyoxometalates through bond order and bond energy analysis. AB - The molecular and electronic structures of transition metal complexes, [MOCl5]n- (n = 2 for M = V,Nb,Ta and n = 1 for Mo,W) and mixed-metal polyoxometalates, [M'M5O19]3-V,Nb,Ta, M = Mo,W) containing a single terminal oxo group on each metal, and of complexes of the uranyl ion [UO2]2+, [UO2(H2O)5]2+ and [UO2Cl4]2-, have been calculated using density functional methods. The calculated structures of the complexes are in good agreement with available experimental parameters. For the mixed-metal hexametalates, for which no crystallographic data is available, the calculations predict a small tetragonal compression of the clusters with only minor structural changes compared to the parent molybdate and tungstate. The metal oxygen bonding in these anions has been probed using Mayer Mulliken, bond energy and atoms in molecule analyses (AIM). These methods provide a consistent description of the bonding in polyoxometalates. The terminal bonds between transition metal or uranium and oxygen atoms have large sigma and pi components with the pi contributions exceeding the sigma bonding. The transition metals utilize their d orbitals almost exclusively to bond to oxygen whilst uranium uses both its 5f and 6d orbitals. Oxygen atom charges increase and covalency indexes decrease with coordination number, with a marked separation of these terms according to the oxygen atom type. The total valency and AIM energies of the oxygen atoms are predicted to be almost constant for all types of oxygen site. The constancy of the bonding power of the oxygen atoms appears to be an important factor in determining the gross structures and details of the bonding in polyoxometalates. The Mayer Mulliken approach provides direct characterization of the bonding power of atoms and the extent of the interaction between pairs of atoms that is consistent with the results of the considerably more computationally demanding bond energy and AIM approaches. PMID- 14527220 TI - Ligand field spectroscopy of Cu(II) and Ag(II) complexes in the gas phase: theory and experiment. AB - Ligand field spectra have been recorded in the gas phase for the two series of complexes containing either Cu(II) or Ag(II) in association with pyridine. Where comparisons are possible, the gas phase spectra match those recorded in the condensed phase; however, for Ag(II) systems the results differ in interpretation. The Ag(II) data are attributed to a ligand-to-metal charge transfer process, and the Cu(II) data (spectral region and extinction coefficient) match the characteristics of a d-d transition. A detailed theoretical analysis of two complexes. [Cu(py)4]2+ and [Ag(py)4]2+ provides evidence of a minimum energy, D4h structure and two less stable D2h and D2d structures within approximately 60 kJ mol(-1). From these structures it is possible to identify a range of optically and vibronically allowed transitions that could contribute to spectra observed in the gas phase. In the case of calculations on [Ag(py)4]2+ there is strong evidence of an electronic transition that would account for the observation of charge transfer in the experiments. Less detailed calculations on [Cu(py)6]2+ and [Ag(py)6]2+ show structural evidence of extensive Jahn Teller distortion. Taken together with incremental binding energies calculated for complexes containing between two and six pyridine molecules, these results show that the level of theory adopted is capable of providing a semi-quantitative understanding of the experimental data. PMID- 14527221 TI - Quantum chemical studies of redox-active enzymes. AB - In applications on mechanisms for metalloenzymes, the hybrid density functional method B3LYP has been used in most cases. The present knowledge about the accuracy of this method on transition metal complexes is described. In comparison to ab initio methods like CASPT2 and CCSD(T), B3LYP has generally been shown to perform quite well. However, there is one exception and this is for the type of copper dimer which is found in enzymes like hemocyanin, catechol oxidase and tyrosinase. Large deviations have been found between CASPT2 and B3LYP and also between B3LYP and experiments on model complexes. This situation is carefully investigated. The accuracy of B3LYP for the activation of O2 in enzymes is also analyzed by comparisons to experiments. For the barrier of O-O bond cleavage B3LYP appears to behave quite satisfactorily, while for the binding of O2 to the metal there are deviations compared to experiment. The question whether this is due to the B3LYP method or to the chemical models used is addressed. PMID- 14527222 TI - Theoretical investigations of Ferredoxin I: the possible role of internal water molecules on the coupled electron proton transfer reaction. AB - The electron-coupled proton transfer reaction involving the [3Fe-4S] cluster in Ferredoxin I is studied by ab initio calculations and molecular dynamics simulations. The charge distributions of the [3Fe-4S] cluster are calculated with density functional theory (B3LYP) and used in the dynamics simulations. Structural differences between the oxidized and reduced clusters in the absence and presence of the protein are calculated to examine the hypothesis that an entatic state is involved in the reaction. The possible role of internal water molecules in the proton transfer process is explored. It is shown that water molecules are dynamically stable near the [3Fe-4S] cluster for tens of ps. Calculations for the native protein and a mutant D15N in which the Asp15 is replaced by a Glu15 are compared. It is found that water is less likely to escape from the region around the [3Fe-4S] cluster in the case of the D15N mutant than in the native protein. This finding could explain, in part, the lower proton transfer rate constant experimentally observed for the mutant if a water molecule were involved in transferring the proton from D15 to the [3Fe-4S] cluster. PMID- 14527223 TI - The rule breaking Cr2(CO)10. A 17 electron Cr system or a Cr=Cr double bond? AB - Density functional theory (DFT) has been used to investigate the conformations and thermochemistry on the singlet and triplet potential energy surfaces (PES) of Cr2(CO)10. The global minimum energy structure for the lowest singlet state of C2h symmetry is consistent with a model of two interacting Cr(CO)5 fragments in which one carbonyl in each fragment acts as an asymmetric four-electron donor bridging carbonyl, with chromium-chromium distances of 2.93 A (B3LYP) or 2.83 A (BP86). Avoiding a Cr...Cr bond by incorporating four-electron donor CO groups in this way allows each chromium atom in singlet Cr(CO)10 to attain the favored 18 electron configuration by using, in a simple picture of the bonding, only the six octahedral sp3d2 hybrids. The dissociation energy to two Cr(CO)5 fragments or to Cr(CO)6 + Cr(CO)4 fragments is predicted to be 10 kcal mol(-1). The lowest triplet state of Cr2(CO)10 is predicted to lie approximately 10 kcal mol(-1) above the singlet global minimum. In the case of triplet Cr2(CO)10 the lowest energy minima were found to be of C2 and C2h symmetry, with similar energies. The chromium-chromium distances in triplet Cr2(CO)10 were found to be shorter than those in the corresponding singlet structures, namely 2.81 (B3LYP) or 2.68 A (BP86) suggesting a sigma + 2(1/2) pi Cr=Cr double bond similar to the O=O bond in O2 or the Fe=Fe bond in the experimentally observed triplet state (Me5C5)2Fe2(mu-CO)3. PMID- 14527224 TI - Dinitrogen activation in sterically-hindered three-coordinate transition metal complexes. AB - Dinuclear metal systems based on sterically-hindered, three-coordinate transition metal complexes of the type ML3 where the ancillary ligands L comprise bulky organic substituents, hold great promise synthetically for the activation and scission of small, multiply-bonded molecules such as N2, NO and N2O. In this study we have employed density functional methods to identify the metal/ligand combinations which achieve optimum activation and/or cleavage of N2. Strong pi donor ligands such as NH2 and OH are found to produce the greatest level of activation based on N-N bond lengths in the intermediate dimer complex, L3Mo(mu N2)MoL3, whereas systems containing the weak or non-pi donor ligands NH3, PH3, OH2 and SH2 are found to be thermodynamically unfavourable for N2 activation. In the case of the Mo-NH2 and W-NH2 systems, a fragment bonding analysis reveals that the orientation of the amide ligands around the metal is important in determining both the spin state and the extent of dinitrogen activation in the intermediate dimer. For both systems, an intermediate dimer structure where one of the NH2 ligands on each metal is rotated 90 degrees relative to the other ligands, is more activated than the structure in which the NH2 ligands are trigonally disposed around the metals. The level of activation is found to be very sensitive to the electronic configuration of the metal with d3 metal ions delivering the best activation along any one transition series. In particular, strong activation or cleavage of N2 was calculated for the third row d3 metals systems involving Ta(II), W(III) and Re(IV), with the level of activation decreasing as the nuclear charge on the metal increases. This trend in activation reflects the size of the valence 5d orbitals and consequently, the capacity of the metal to back donate into the dinitrogen pi* orbitals. PMID- 14527225 TI - The full topology of the Laplacian of the electron density: scrutinising a physical basis for the VSEPR model. AB - Within the framework of quantum chemical topology (QCT) the function L(r), which equals the negative of the Laplacian of the electron density, has been proposed before as a physical basis for the valence shell electron pair repulsion (VSEPR) model. The availability of a new algorithm to integrate property densities over the basins of L(r) enabled a re-evaluation of this physical basis. We optimised a set of nine molecules at B3LYP/6-311+G(2d,p) level and partitioned the corresponding L(r) function for each molecule into basins. For the first time we visualise these basins in L(r), by directly showing their boundaries. We identify the basins in L(r) with the domains of the VSEPR model. Observations drawn from the populations and volumes of L-basins are contrasted with the three subsidiary VSEPR postulates. We find unexpectedly small populations, nearer to one than to two, for non-hydrogen cores and bonding domains, and populations much larger than two for non-bonding domains. We conclude that non-bonding or lone pairs have larger domains than bonding pairs in the same valence shell, in accordance with VSEPR. We also confirm that double and triple bond domains are larger than single bond domains. However we cannot substantiate the effect of the electronegativity of central atom or ligand on the volume of bonding domains. In summary, the full topology of L(r) supports two out of three subsidiary VSEPR postulates. PMID- 14527226 TI - Energy decomposition analysis of the chemical bond in main group and transition metal compounds. AB - The nature of the chemical bond in the main group diborane(4) compounds X2B-BX2 (X = H,F,Cl,Br,I) and in the Fischer- and Schrock-type transition metal carbene and carbyne complexes and heavier homologues (CO)5W-CH2, (CO)5W-E(OH)2, Cl4W-EH2, Cl(CO)4W-EH and Cl3W-EH (E = C,Si,Ge,Sn,Pb) have been investigated with an energy decomposition analysis (EDA). The results give a deep insight into the nature of the chemical interactions. The EDA results can be used as a bridge between the heuristic models of experimental chemists which have been proven as useful ordering schemes for experimental observations and the physical mechanism which leads to a chemical bond. At the time the data give a well defined qantitative answer to the questions about the strength of the covalent and electrostatic interactions and about the contributions of sigma and pi electrons to the covalent bond. PMID- 14527227 TI - Ligand field and density functional descriptions of the d-states and bonding in transition metal complexes. AB - The d-orbital energy sequences for low symmetry transition metal complexes derived from Kohn-Sham density functional theory and ligand field theory are different due to each model's treatment of interelectron repulsion. The implications for providing a unified description of the underlying metal-ligand bonding are analysed and illustrated using conventional and time-dependent DFT. Previous detailed spectroscopic studies have established the d orbital sequence in planar coordination complexes containing pi-donor halide ligands as dx2-y2 >> dxy > dxz, dyz > dz2 while for a sigma-only system like [Pd(NH3)4]2+, ligand field approaches like the angular overlap model (AOM) or cellular ligand field (CLF) model predict dxz, dyz, and dxy, should be degenerate. However, the energies of the Kohn Sham (KS) 'd' orbitals of [PdCl4]2- and [Pd(NH3)4]2+ place dxy below the dxz/dyz pair. Direct use of the KS orbital eigenvalues in AOM or CLF analyses would imply both ligands are pi acceptors. This result is independent of the choice of functional or whether the calculation is carried out in vacuo or in a polarised continuum representing solvation by water. The origin of the difference between the KS and LFT d orbital sequences derives from their treatments of d-d interelectron repulsion. KS orbitals include interelectron repulsion contributions while LFT d orbitals do not. For a low-spin d8 complex, DFT gives less d-d interelectron repulsion in the xy plane leading to a lowering of dxy relative to dxy/dyz. This differential effect can be reversed qualitatively by progressively removing electron density from dz2 and placing it in dx2-y2. When about 0.6 electrons is rearranged, E(dxy) > E(dxz/dyz) for [PdCl4]2- and the dpi orbitals for [Pd(NH3)4]2+ are virtually degenerate. The wider ramifications of these interelectron repulsion effects are discussed for other symmetries. Excited d state energies for [PdCl4]2- are computed using both time dependent density functional theory (TDDFT) and determinant energies. The latter give the experimental sequence 1A2g < 1Eg < 1B1g while TDDFT gives 1Eg < 1A2g < 1B1g. Both give transition energies up to 30% lower than observed. A DFT analysis of the bonding energies in [PdCl4]2- indicates that the Pd-Cl pi bonding in the molecular plane is about 33% weaker than the out-of-plane pi interaction due to non-zero overlap between ligand orbitals. The normal LFT assumption of linear ligation may not always be valid. PMID- 14527228 TI - Are current DFT methods sufficiently reliable for real-world molecular systems? AB - Whereas density functional theory (DFT) based computations are the main tools in contemporary computational chemistry, there is an issue of chemical accuracy and reliability which, one the one hand, is recognised but, on the other hand, is not always sufficiently dealt with when it comes to application. Examples are given where DFT apparently fails. It is suggested that an effort needs to be undertaken to thoroughly investigate the limitations of current DFT calculations. PMID- 14527229 TI - The use of methods involving semi-empirical molecular orbital theory to study the structure and reactivity of transition metal complexes. AB - The electronic structure of molecular systems containing transition metal atoms is traditionally studied using methods based on density functional theory (DFT). Although such an approach has been quite successful, the treatment of large systems, be they transition metal complexes, bioinorganic molecules or the solid state, is still extremely computationally demanding at this level, and may not be practical for many systems of interest. In this paper we describe how semi empirical MO methods can be used to overcome these computational bottlenecks, yet still provide predictions of the necessary accuracy. We describe strategies to achieve this by focussing on: (i) obtaining appropriate parameters for transition metal atoms via a genetic algorithm, to be used within a parallelised implementation of neglect of differential diatomic overlap (NDDO) methods, and (ii) the use of multilevel treatments which involve DFT and semi-empirical methods to describe different regions of the molecule. Here we show by reference to histidine and porphyrin complexes, the importance of a correct partitioning of the organic substrate. We illustrate the potential of such a dual level approach by reporting preliminary results showing the catalytic role of the enzyme, dimethyl sulfoxide reductase. PMID- 14527230 TI - Some critical issues in the application of quantum mechanics/molecular mechanics methods to the study of transition metal complexes. AB - The application of quantum mechanics/molecular mechanics (QM/MM) methods in transition metal chemistry is growing steadily. It becomes therefore appropriate to assess the importance of a number of technical issues associated to their implementation. This work presents the discussion of several of these issues, including the eventual need for conformational searches, the choice of the MM force field and the possibility of its tuning. The examples presented here prove that a proper handling of these technical aspects can lead to an improvement in the efficiency and quality of QM/MM calculations. PMID- 14527231 TI - Escalating health care spending: is it desirable or inevitable? AB - This study analyzes changing trends in U.S. health spending and concludes that although the long-term growth trend has been a good predictor of future spending, periodic differences in the growth trend are important. Of particular concern is the rapid acceleration in health spending beginning in 1998. If left unchecked, the current growth rate will result in almost 24 percent of GDP spent on health by 2011. The authors question whether such unconstrained spending levels are either desirable or inevitable, and they offer a guide to how the United States might develop a long-term cost-containment strategy that is both effective and sustainable. PMID- 14527232 TI - Should we be worried about high real medical spending growth in the United States? AB - This paper argues that increases in medical spending growth are not necessarily causes for alarm or reasons for strong action. Especially in the private sector, increases in employers' payments for benefits should often be offset with smaller raises; whether employees' cost shares should be increased depends on a comparison of benefits and costs. Problems are more severe for public-sector spending, for the uninsured, and for those responding to distorted incentives. PMID- 14527233 TI - Should public policy seek to control the growth of health care spending? AB - Debate about whether to restrain health care spending is intensifying. Much of the debate revolves around efficient research allocation. The possible impact of cost control on the advance of medical technology merits more attention. Public and private efforts to slow growth of health spending have not enjoyed much success. Most recently, managed care failed because administrators lacked political legitimacy. Politically established budget limits can hold down spending, but the United States seems unwilling to adopt them; even if it were, the technology to administer them rationally does not exist. In this situation, efforts to hold down cost growth carry grave risks. PMID- 14527234 TI - Confronting the barriers to chronic care management in Medicare. AB - This paper examines the ability of the current Medicare program--both traditional fee-for-service and risk-based contracting--to address the needs of beneficiaries with chronic conditions, who represent almost 80 percent of program enrollment. Grounded in indemnity insurance principles, including concerns about "moral hazard," the traditional Medicare program faces difficulty evolving to support of a chronic care model of health care practice. Although capitation may be the most desirable platform to support provision of care to beneficiaries with chronic conditions, the current structural limitations and problems faced in the Medicare+Choice program limit capitation's use at this time. PMID- 14527235 TI - Health spending projections for 2002-2012. AB - We forecast a slowdown in national health spending growth in 2002 and 2003, reflecting slower projected Medicare and private personal health spending growth. These factors outweigh higher projected Medicaid spending growth, caused by weak labor markets, and an expectation of continued high private health insurance premium inflation related to the underwriting cycle. Over the entire projection period, national health spending growth is still expected to outpace economic growth. The result is that the health share of gross domestic product is projected to increase from 14.1 percent in 2001 to 17.7 percent in 2012. PMID- 14527236 TI - How much medical care do the uninsured use, and who pays for it? AB - With the number of uninsured people exceeding forty-one million in 2001, insuring the uninsured is again a major policy issue. This analysis establishes benchmarks for the inevitable debate over the cost of expanding coverage: How much is being spent on care for the uninsured, and where does the money come from? This information is essential for assessing how much new money will be required for expanded coverage, how much can be reallocated from existing sources, and how a new financing system would redistribute the burden of subsidizing care for the uninsured from private to public sources. PMID- 14527237 TI - Consumers' reports on the health effects of direct-to-consumer drug advertising. AB - We conducted a national telephone survey about health care experiences associated with direct-to-consumer advertising (DTCA) of prescription drugs. Among the 35 percent of our sample who had a physician visit during which DTCA was discussed, 25 percent received a new diagnosis, of which 43 percent were considered high priority according to authoritative sources. More than half also reported actions taken by their physician other than prescribing the advertised drug. Despite concerns about DTCA's negative consequences, we found no differences in health effects between patients who took advertised drugs and those who took other prescription drugs. PMID- 14527238 TI - Pharmaceutical promotion: don't throw the baby out with the bathwater. AB - Spending on prescription drugs and promotion by the pharmaceutical industry grew substantially during the past ten years. Does the greater exposure offered by promotion fill a needed educational gap, or does it merely promote inappropriate use? This paper uses two recent studies to explore this question, presenting a framework in which the impact of promotion depends upon the level of evidence and consensus on drug use. PMID- 14527239 TI - Advertising and prescription drugs: promotion, education, and the public's health. AB - The data presented by Joel Weissman and colleagues and by Robert Dubois do not justify the conclusions that the effects of pharmaceutical promotion are beneficial. Among consumers of direct-to-consumer advertising (Weissman and colleagues), those heavily influenced by such promotion were no more likely than others were to have new conditions diagnosed or confirmed and were much less likely to have laboratory studies ordered or lifestyle changes recommended. A second study (Dubois) arguing that drug advertising improves the appropriateness of prescribing relies on unconvincing ecological arguments. A greater presence of noncommercial, public health-oriented communication would make a more useful and cost-effective impact on the nation's health. PMID- 14527240 TI - Pharmaceutical promotion, advertising, and consumers. AB - Proponents of drug promotion and advertising claim that it is informative and educational; opponents are concerned that the information conveyed encourages inappropriate and unnecessary use. Health Affairs papers by Joel Weissman and colleagues and by Robert Dubois provide some validation for the views of both sides of this debate but do not allow us to draw definitive conclusions about key issues involving inappropriate use of expensive medications and their substitution for cheaper medications that are just as effective. The extent to which consumers have been protected from the rising cost of pharmaceuticals further muddles the picture. However, new insurance benefit designs that threaten to shift more costs to consumers may fuel demand for more comprehensive and balanced information. PMID- 14527241 TI - Two advertisements for TV drug ads. AB - The paper by Joel Weissman and colleagues addresses the increasingly important topic of the effects of direct-to-consumer advertising (DTCA) by pharmaceutical companies. The authors claim that their results should be reassuring to "those concerned about potential adverse health care consequences of DTCA". However, the study and analysis of the data are marred by several flaws that diminish the importance and relevance of the findings, including weakness in design, overgenerous interpretations, and failure to address key questions. Rather than informing the debate, the study amounts to little more than an advertisement for drug advertisements. PMID- 14527242 TI - What do we know about direct-to-consumer advertising of prescription drugs? AB - Two papers, by Joel Weissman and colleagues and by Robert Dubois, add to our limited knowledge of the effects of direct-to-consumer (DTC) advertising of prescription drugs. Their results reinforce the largely positive findings from consumer surveys, while adding valuable new data and insights. These suggest that DTC ads probably improve patients' health outcomes and do not tend to lead to inappropriate prescribing. DTC advertising is emerging as a positive force in health care markets, consistent with what is known about the effects of advertising in many other markets. PMID- 14527243 TI - Examining the FDA's oversight of direct-to-consumer advertising. AB - Our analysis examined the effects of the Food and Drug Administration's (FDA's) 1997 draft guidance regarding advertisements for prescription drugs broadcast directly to consumers. We found that although direct-to-consumer (DTC) advertising spending by pharmaceutical companies has increased, more than 80 percent of their promotional spending is directed to physicians. DTC advertising appears to increase the use of prescription drugs among consumers. The FDA's oversight has not prevented companies from making misleading claims in subsequent advertisements, and a recent policy change has lengthened the FDA's review process, raising the possibility that some misleading campaigns could run their course before review. PMID- 14527244 TI - Thinking outside the box: a conversation with John Breaux. Interview by Gail R. Wilensky. PMID- 14527245 TI - The Breaux Plan: a radically centrist approach to a new health care system. AB - Sen. John Breaux (D-LA) proposes a new universal health insurance scheme that takes the best from both the private and the public sectors. Government would use its strength as regulator to set minimum standards and offer a fallback coverage system; states would organize their own insurance pools and arrange coverage for the most vulnerable. The private sector would play a critical role to ensure that the new system was based on principles of competition; employers would have the option of covering their workers much as they do now; and individual coverage would still be available for those who desire and can afford it. For the first time, Americans would be held accountable for obtaining health insurance and would be assisted in doing so. PMID- 14527246 TI - Hospital tiers in health insurance: balancing consumer choice with financial incentives. AB - Variations in efficiency and market power are generating wide variations in the prices charged by hospitals to health insurance plans. Insurers are developing new network structures that expose the consumer to some of the cost differences, to encourage but not mandate differential use of the more economical facilities. The three leading designs include hospital "tiers" within a single broad network, multiple-network products, and the replacement of copayments by coinsurance in HMO as well as PPO products. This paper describes the new network designs and evaluates the challenges they face in influencing consumers' behavior, incorporating information on clinical quality, and supporting medical education and uncompensated care. PMID- 14527247 TI - Tiered hospital networks. AB - As a result of rising health care costs, health plans are experimenting with insurance products that shift greater financial responsibility for medical care to consumers and create incentives for consumers to consider cost differences when choosing among providers. Based on an October 2002 roundtable discussion, this paper discusses insurance product trends, particularly tiered hospital networks. Issues addressed include these product features' potential to reduce system costs, the effect on the hospital-health plan relationship, consumers' ability to consider cost and quality in decision making, and financial barriers to care for the chronically ill. PMID- 14527248 TI - Hospital tiering: how will it play in Peoria? AB - Cost-sharing strategies such as hospital tiering will require consumers to make cost-benefit decisions where they have little experience. This responsibility may be further challenged by prevailing consumer perspectives: that health insurance is an open-ended service benefit; that medical treatment decisions should not be influenced by costs; and that consumers are not responsible for the current cost crisis. Although there are steps providers can take to prepare consumers for their new role in cost sharing, health care leaders need to begin moving from a consumer-driven to a citizen-driven approach. PMID- 14527249 TI - The erosion of health insurance: the unintended consequences of tiered products by health plans. AB - When properly structured, consumer-driven health care may provide gains to both patients and the delivery system. However, the current approach by health plans could result in real harm to patients and to an already fragile health care delivery system. While health plans are presenting tiered products as a necessary mechanism to control rising hospital expenditures, this paper explores the real drivers of the rising cost of health care, including utilization, increased demand for advanced medication, and new technology. Left unchecked, such benefit designs could have dangerous public policy implications and consequences, including the further erosion of the basic tenets of health insurance. PMID- 14527250 TI - Improving the quality of Medicaid personal assistance through consumer direction. AB - As states seek to improve home and community-based services for people with disabilities, many are incorporating consumer-directed supportive services into their Medicaid programs. The national Cash and Counseling Demonstration uses a randomized design to compare an innovative model of consumer direction with the traditional agency-directed approach. This paper presents findings from the first demonstration program to be implemented, in Arkansas. Our survey of 1,739 elderly and nonelderly adults showed that relative to agency-directed services, Cash and Counseling greatly improved satisfaction and reduced most unmet needs. Moreover, contrary to some concerns, it did not adversely affect participants' health and safety. PMID- 14527251 TI - Can managed care and competition control Medicare costs? AB - Medicare+Choice (M+C) was conceived to bring managed care and competitive forces to bear on Medicare. Ultimately, M+C could not thrive under the conditions of the marketplace and the Balanced Budget Act of 1997. Here I review what went wrong and the lessons from the experience, concluding that M+C is a tool, not a strategy. While managed care in a multiple-choice environment may have the potential to generate limited savings, promoting managed care and competition alone will not preempt the need for a debate on Medicare's obligations and how to finance them. PMID- 14527252 TI - Health insurance and spending among cancer patients. AB - Over 1.3 million new cancer cases are diagnosed each year. While most cancer patients are older and covered by Medicare, our analysis indicates that 10 percent of cancer patients under age sixty-five are uninsured and that 20 percent of Hispanic cancer patients under age sixty-five are uninsured. We find substantial differences in cancer spending by insurance status; uninsured patients under age sixty-five spent 57 percent as much over a six-month period as privately insured patients spent for their cancer care. We present evidence to show that spending differences are due in part or completely to differences in use, which suggests that raising coverage rates will improve cancer treatment. PMID- 14527253 TI - Creating consensus on coverage choices. AB - The framework for reaching near-universal coverage outlined in this paper combines tax credits for private insurance and public program expansions. It illustrates how a series of incremental steps could be phased in to achieve near universal coverage. Hallmarks include creation of a Congressional Health Plan; use of the income tax system to provide tax credits and enroll uninsured people; creation of a state Family Health Insurance Program open to everyone below 150 percent of poverty; and creation of a Medicare Part E, open to the disabled and uninsured older adults. The paper provides coverage and cost estimates and identifies potential sources of revenue to finance coverage. PMID- 14527254 TI - Reforming health insurance: realistic options and hard choices. AB - The persistently high number of people reporting that they have no health insurance has precipitated a number of new comprehensive proposals to extend coverage to most Americans. Such proposals must find solutions to fundamental problems that have thus far eluded policymakers, including the role of government regulation, how much to spend, and who should pay. The experience of the 1990s suggests that an effective policy would offer more choices of health plans through the private market, limit unnecessary government regulation, and provide appropriate subsidies to individuals. PMID- 14527255 TI - A relevant universal coverage proposal. AB - Karen Davis and Cathy Schoen offer a strategic vision for universal coverage that attempts to move beyond ideological battles that have stifled progress. However, I believe that there are a few specific shortcomings with the proposal's logic that could thwart political consensus. I review some of these shortcomings and make suggestions for incremental technical improvements. In particular, I suggest that future versions of the proposal consider administering the tax credits as a "passthrough" from the government through employers to individuals. I also believe that reform proposals should address more directly the issues of provider accountability and patient information needs. PMID- 14527256 TI - National medical spending attributable to overweight and obesity: how much, and who's paying? AB - We use a regression framework and nationally representative data to compute aggregate overweight- and obesity-attributable medical spending for the United States and for select payers. Combined, such expenditures accounted for 9.1 percent of total annual U.S. medical expenditures in 1998 and may have been as high as dollar 78.5 billion (dollar 92.6 billion in 2002 dollars). Medicare and Medicaid finance approximately half of these costs. PMID- 14527257 TI - Use of subsidies to low-income people for coverage through small employers. AB - If tax credits or other public subsidies are made available only for health insurance that is not employment-based, serious erosion of employer coverage could result. To prevent this, public subsidies targeted to low-income workers and families could be applied in ways that broaden employer coverage for low income workers and their families by encouraging small employers with largely low wage workforces to offer and partially fund health coverage for their workers. To accomplish this, such employers--very few of which now provide health coverage- must be allowed to contribute much less than normally required in the commercial market. PMID- 14527258 TI - Employment-based health insurance is failing: now what? AB - Employment-based health insurance is failing. Costs are out of control. Employers have no effective strategy to deal with this. They must think strategically about fundamental change. This analysis explains how employers' purchasing policies contribute to rising costs and block growth of economical care. Single-source managed care is ineffective, and effective managed care cannot be a single source. Employers should create exchanges through which they can offer employees wide, responsible, individual, multiple choices among health care delivery systems and create serious competition based on value for money. Recently introduced technology can assist this process. PMID- 14527259 TI - Covering the uninsured: how much would it cost? AB - To provide benchmarks for evaluating the costs of alternative proposals to provide insurance coverage for the uninsured, this study presents two sets of cost estimates derived from medical spending patterns of lower- or middle-income people with private insurance plans and those of people with public insurance coverage during 1996-1998. The analysis suggests that the uninsured would use dollar 33.9-dollar 68.7 billion (in 2001 dollars) in additional medical care if they were fully insured. An increase in medical spending of this range would increase total health care spending by 3-6 percent and would raise health care's share of GDP by less than one percentage point. PMID- 14527260 TI - Tracking health care costs: trends stabilize but remain high in 2002. AB - Health care spending per privately insured person increased 9.6 percent in 2002, a slight reduction from the 10 percent increase in 2001. This is the first time in five years that the spending trend did not accelerate. Nonetheless, health care spending grew nearly four times faster than the U.S. economy grew in 2002. Growth in hospital spending accounted for the largest portion of the overall increase (51 percent) for the second straight year. Moreover, hospital price inflation--which accelerated significantly in 2002--accounted for a larger share of hospital spending growth in 2002 than in 2001. Premium increases accelerated again in 2003, despite 2002's slight deceleration of the overall spending trend. PMID- 14527261 TI - Managing care: utilization review in action at two capitated medical groups. AB - Despite widespread concern about denials of coverage by managed care organizations, little empirical information exists on the profile and outcomes of utilization review decisions. This study examines the outcomes of nearly a half million coverage requests in two large medical groups that contract with health plans to deliver care and conduct utilization review. We found much higher denial rates than those previously reported. Denials were particularly common for emergency care and durable medical equipment. Retrospective requests were nearly four times more likely than prospective requests were to be denied, and when prospective requests were denied, it was more likely because the service fell outside the scope of covered benefits than because it was not medically necessary. PMID- 14527262 TI - AHCPR and the changing politics of health services research. AB - The Agency for Health Care Policy and Research has had a turbulent history. Created with little opposition in 1989, it narrowly escaped being eliminated in 1995, only to be reauthorized (with a new mandate and name--the Agency for Healthcare Research and Quality, or AHRQ) with overwhelming support in 1999. In focusing on budgetary history, this paper sheds light on why health services research (HSR) has difficulty obtaining funding from a government that is willing to spend vast sums on basic biomedical research. The paper argues that three strategies--bureaucratic, marketing, and constituency building--that advocates adopted in the late 1980s made HSR more visible and consequential and were responsible for AHCPR's budgetary successes as well as its near-demise. PMID- 14527263 TI - The more things change...: the federal government's role in the evaluative sciences. AB - The unfortunate political history of the Agency for Health Care Policy and Research (AHCPR) illustrates the risks to the agencies attempting to evaluate the common practices of medicine and reform clinical decision making to take account of patients' preferences. The evaluative sciences have yet to regain the congressional attention they had when Senators George Mitchell and David Durenberger championed their cause. But the fundamental problems remain, and they are getting worse. Sooner or later Congress will need to revisit the debate over where in the federal government the evaluative sciences should find their base, and questions concerning the role of the National Institutes of Health (NIH) will be raised once again, as they were at the time of AHCPR's founding. PMID- 14527264 TI - An insider's perspective on the near-death experience of AHCPR. AB - The story of AHCPR's struggle with Congress to remain a federal agency is an example of the longer struggle health services research (HSR) has had to hold a priority in the federal budget. But from another perspective, the growth of HSR has been substantial, albeit an up-and-down experience. Its survival during the turbulent years of 1995-1996 is attributed to a fundamental restructuring of its program priorities and building a new base of support from major health care associations and leaders. A fortunate sequence of events also contributed to the reversal of what could have been a cataclysmic occurrence for the field of HSR. PMID- 14527265 TI - Back to the future. AB - The paper by Brad Gray and colleagues summarizes a decade of challenge, growth, and evolution within what is now called the Agency for Healthcare Research and Quality (AHRQ) and the field of health services research, and it gives new depth to the old saying, "May you live in interesting times". Their assessment of the significance of the agency's reauthorization and description of continued challenges for AHRQ and the field are insightful. This commentary focuses on continued maturation of AHRQ's mission and focus, recent achievements, new external factors, and emerging policy dilemmas that AHRQ is uniquely poised to address. PMID- 14527266 TI - Coding of auditory space. AB - Behavioral, anatomical, and physiological approaches can be integrated in the study of sound localization in barn owls. Space representation in owls provides a useful example for discussion of place and ensemble coding. Selectivity for space is broad and ambiguous in low-order neurons. Parallel pathways for binaural cues and for different frequency bands converge on high-order space-specific neurons, which encode space more precisely. An ensemble of broadly tuned place-coding neurons may converge on a single high-order neuron to create an improved labeled line. Thus, the two coding schemes are not alternate methods. Owls can localize sounds by using either the isomorphic map of auditory space in the midbrain or forebrain neural networks in which space is not mapped. PMID- 14527267 TI - Novel neural modulators. AB - The discovery that nitric oxide (NO) is produced by neurons and regulates synaptic activity has challenged the definition of a neurotransmitter. NO is not stored in synaptic vesicles and does not act at conventional receptors on the surface of adjacent neurons. The toxic gases carbon monoxide (CO) and hydrogen sulfide (H2S) are also produced by neurons and modulate synaptic activity. D serine synthesis and release by astrocytes as an endogenous ligand for the "glycine" site of N-methyl D-aspartate (NMDA) receptors defy the concept that a neurotransmitter must be synthesized by neurons. We review the properties of these "atypical" neural modulators. PMID- 14527268 TI - The neurobiology of visual-saccadic decision making. AB - Over the past two decades significant progress has been made toward understanding the neural basis of primate decision making, the biological process that combines sensory data with stored information to select and execute behavioral responses. The most striking progress in this area has been made in studies of visual saccadic decision making, a system that is becoming a model for understanding decision making in general. In this system, theoretical models of efficient decision making developed in the social sciences are beginning to be used to describe the computations the brain must perform when it connects sensation and action. Guided in part by these economic models, neurophysiologists have been able to describe neuronal activity recorded from the brains of awake-behaving primates during actual decision making. These recent studies have examined the neural basis of decisions, ranging from those made in predictable sensorimotor tasks to those unpredictable decisions made when animals are engaged in strategic conflict. All of these experiments seem to describe a surprisingly well integrated set of physiological mechanisms that can account for a broad range of behavioral phenomena. This review presents many of these recent studies within the emerging neuroeconomic framework for understanding primate decision making. PMID- 14527269 TI - Generating the cerebral cortical area map. AB - The view that the cortical primordium is initially patterned in similar ways to the rest of the embryo has been a conceptual breakthrough. We now have a new starting point for understanding how the cortical area map is established and how maps may change and evolve. Here we review findings that signaling molecules secreted from distinct cortical signaling centers establish positional information in the cortical primordium and regulate regional growth. In other embryonic systems, positional signals would regulate the patterned expression of transcription factors, leading, in a gene regulatory cascade, to the patterned differentiation of the tissue. We discuss candidate transcription factors with respect to such a model of cortical patterning. Finally, embryonic structures interact to pattern one another. We review data suggesting that the thalamus and cortex are patterned independently then interact to generate the final cortical area map. PMID- 14527270 TI - The biology of epilepsy genes. AB - Mutations in over 70 genes now define biological pathways leading to epilepsy, an episodic dysrhythmia of the cerebral cortex marked by abnormal network synchronization. Some of the inherited errors destabilize neuronal signaling by inflicting primary disorders of membrane excitability and synaptic transmission, whereas others do so indirectly by perturbing critical control points that balance the developmental assembly of inhibitory and excitatory circuits. The genetic diversity is now sufficient to discern short- and long-range functional convergence of epileptogenic molecular pathways, reducing the broad spectrum of primary molecular defects to a few common processes regulating cortical synchronization. Synaptic inhibition appears to be the most frequent target; however, each gene mutation retains unique phenotypic features. This review selects exemplary members of several gene families to illustrate principal categories of the disease and trace the biological pathways to epileptogenesis in the developing brain. PMID- 14527271 TI - Progress toward understanding the genetic and biochemical mechanisms of inherited photoreceptor degenerations. AB - More than 80 genes associated with human photoreceptor degenerations have been identified. Attention must now turn toward defining the mechanisms that lead to photoreceptor death, which occurs years to decades after the birth of the cells. Consequently, this review focuses on topics that offer insights into such mechanisms, including the one-hit or constant risk model of photoreceptor death; topological patterns of photoreceptor degeneration; mutations in ubiquitously expressed splicing factor genes associated only with photoreceptor degeneration; disorders of the retinal pigment epithelium; modifier genes; and global gene expression analysis of the retina, which will greatly increase our understanding of the downstream events that occur in response to a mutation. PMID- 14527272 TI - Cell biology of the presynaptic terminal. AB - The chemical synapse is a specialized intercellular junction that operates nearly autonomously to allow rapid, specific, and local communication between neurons. Focusing our attention on the presynaptic terminal, we review the current understanding of how synaptic morphology is maintained and then the mechanisms in synaptic vesicle exocytosis and recycling. PMID- 14527273 TI - Gathering no moss. AB - I never imagined that I would be asked to write an autobiography in a microbiology tome. For that matter, little did I think that I would consider microbiology the most intriguing subject in the life sciences and the only field I wanted to study. My formal scientific training was in chemistry. This is a recounting of my conversion and the opportunities I have had to work in the microbial sciences with some of the major figures (and characters) during a period of marvelous intensity and productivity. I want to recognize and thank my many distinguished colleagues for the ways in which they have helped me to experience a fruitful and stimulating life as a microbiologist. PMID- 14527274 TI - Molecular pathogenicity of the oral opportunistic pathogen Actinobacillus actinomycetemcomitans. AB - Periodontitis is mankind's most common chronic inflammatory disease. One severe form of periodontitis is localized aggressive periodontitis (LAP), a condition to which individuals of African origin demonstrate an increased susceptibility. The main causative organism of this disease is Actinobacillus actinomycetemcomitans. A member of the Pasteurellaceae, A. actinomycetemcomitans produces a number of interesting putative virulence factors including (a) an RTX leukotoxin that targets only neutrophils and monocytes and whose action is influenced by a novel type IV secretion system involved in bacterial adhesion; (b) the newly discovered toxin, cytolethal distending toxin (CDT); and (c) a secreted chaperonin 60 with potent leukocyte-activating and bone resorbing activities. This organism also produces a plethora of proteins able to inhibit eukaryotic cell cycle progression and proteins and peptides that can induce distinct forms of proinflammatory cytokine networks. A range of other proteins interacting with the host is currently being uncovered. In addition to these secreted factors, A. actinomycetemcomitans is invasive with an unusual mechanism for entering, and traveling within, eukaryotic cells. This review focuses on recent advances in our understanding of the molecular and cellular pathogenicity of this fascinating oral bacterium. PMID- 14527276 TI - On the trail of a cereal killer: Exploring the biology of Magnaporthe grisea. AB - The blast fungus Magnaporthe grisea causes a serious disease on a wide variety of grasses including rice, wheat, and barley. Rice blast is the most serious disease of cultivated rice and therefore poses a threat to the world's most important food security crop. Here, I review recent progress toward understanding the molecular biology of plant infection by M. grisea, which involves development of a specialized cell, the appressorium. This dome-shaped cell generates enormous turgor pressure and physical force, allowing the fungus to breach the host cuticle and invade plant tissue. The review also considers the role of avirulence genes in M. grisea and the mechanisms by which resistant rice cultivars are able to perceive the fungus and defend themselves. Finally, the likely mechanisms that promote genetic diversity in M. grisea and our current understanding of the population structure of the blast fungus are evaluated. PMID- 14527277 TI - Bacterial membrane lipids: where do we stand? AB - Phospholipids play multiple roles in bacterial cells. These are the establishment of the permeability barrier, provision of the environment for many enzyme and transporter proteins, and they influence membrane-related processes such as protein export and DNA replication. The lipid synthetic pathway also provides precursors for protein modification and for the synthesis of other molecules. This review concentrates on the phospholipid synthetic pathway and discusses recent data on the synthesis and function of phospholipids mainly in the bacterium Escherichia coli. PMID- 14527275 TI - Assembly dynamics of the bacterial cell division protein FTSZ: poised at the edge of stability. AB - FtsZ is a prokaryotic tubulin homolog that assembles into a ring at the future site of cell division. The resulting "Z ring" forms the framework for the division apparatus, and its assembly is regulated throughout the bacterial cell cycle. A highly dynamic structure, the Z ring exhibits continual subunit turnover and the ability to rapidly assemble, disassemble, and, under certain circumstances, relocalize. These in vivo properties are ultimately due to FtsZ's capacity for guanosine triphosphate (GTP)-dependent, reversible polymerization. FtsZ polymer stability appears to be fine-tuned such that subtle changes in its assembly kinetics result in large changes in the Z ring structure. Thus, regulatory proteins that modulate FtsZ's assembly dynamics can cause the ring to rapidly remodel in response to developmental and environmental cues. PMID- 14527278 TI - Spatial and temporal control of differentiation and cell cycle progression in Caulobacter crescentus. AB - The dimorphic and intrinsically asymmetric bacterium Caulobacter crescentus has become an important model organism to study the bacterial cell cycle, cell polarity, and polar differentiation. A multifaceted regulatory network orchestrates the precise coordination between the development of polar organelles and the cell cycle. One master response regulator, CtrA, directly controls the initiation of chromosome replication as well as several aspects of polar morphogenesis and cell division. CtrA activity is temporally and spatially regulated by multiple partially redundant control mechanisms, such as transcription, phosphorylation, and targeted proteolysis. A multicomponent signal transduction network upstream CtrA, containing histidine kinases CckA, PleC, DivJ, and DivL and the essential response regulator DivK, contributes to the control of CtrA activity in response to cell cycle and developmental cues. An intriguing feature of this signaling network is the dynamic cell cycle-dependent polar localization of its components, which is believed to have a novel regulatory function. PMID- 14527279 TI - Bacterial motility on a surface: many ways to a common goal. AB - When free-living bacteria colonize biotic or abiotic surfaces, the resultant changes in physiology and morphology have important consequences on their growth, development, and survival. Surface motility, biofilm formation, fruiting body development, and host invasion are some of the manifestations of functional responses to surface colonization. Bacteria may sense the growth surface either directly through physical contact or indirectly by sensing the proximity of fellow bacteria. Extracellular signals that elicit new gene expression include autoinducers, amino acids, peptides, proteins, and carbohydrates. This review focuses mainly on surface motility and makes comparisons to features shared by other surface phenomenon. PMID- 14527280 TI - Transposable elements in filamentous fungi. AB - The past 10 years have been productive in the characterization of fungal transposable elements (TEs). All eukaryotic TEs described are found including an extraordinary prevalence of active members of the pogo family. The role of TEs in mutation and genome organization is well documented, leading to significant advances in our perception of the mechanisms underlying genetic changes in these organisms. TE-mediated changes, associated with transposition and recombination, provide a broad range of genetic variation, which is useful for natural populations in their adaptation to environmental constraints, especially for those lacking the sexual stage. Interestingly, some fungal species have evolved distinct silencing mechanisms that are regarded as host defense systems against TEs. The examination of forces acting on the evolutionary dynamics of TEs should provide important insights into the interactions between TEs and the fungal genome. Another issue of major significance is the practical applications of TEs in gene tagging and population analysis, which will undoubtedly facilitate research in systematic biology and functional genomics. PMID- 14527281 TI - Bacteriophage-induced modifications of host RNA polymerase. AB - Bacteriophages have developed an impressive array of ingenious mechanisms to modify bacterial host RNA polymerase to make it serve viral needs. In this review we summarize the current knowledge about two types of host RNA polymerase modifications induced by double-stranded DNA phages: covalent modifications and modifications through RNA polymerase-binding proteins. We interpret the biochemical and genetic data within the framework of a structure-function model of bacterial RNA polymerase and viral biology. PMID- 14527282 TI - Vaccinia virus motility. AB - Vaccinia virus (VV), the virus smallpox vaccine, replicates in the cytoplasm of infected cells. The intracellular movement of this large virus would be inefficient without specific transport mechanisms; therefore, VV uses microtubules for movement during both entry and egress. In addition, the dissemination of virus from infected cells to adjacent cells is promoted by the polymerization of actin beneath cell surface virions to drive virus particles away from the cell. Last, the roles of different VV particles in virus movement within and between hosts are discussed. PMID- 14527283 TI - Measles virus 1998-2002: progress and controversy. AB - Despite the extensive media exposure that viruses such as West Nile, Norwalk, and Ebola have received lately, and the emerging threat that old pathogens may reappear as new agents of terrorism, measles virus (MV) persists as one of the leading causes of death by infectious agents worldwide, approaching the annual mortality rate of human immunodeficiency virus (HIV)-1. For most MV victims, fatality is indirect: Virus-induced transient immunosuppression predisposes the individual to opportunistic infections that, left untreated, can result in mortality. In rare cases, MV may also cause progressive neurodegenerative disease. During the past five years (1998-2002), development of animal models and the application of reverse genetics and immunological assays have collectively contributed to major progress in our understanding of MV biology and pathogenesis. Nevertheless, questions and controversies remain that are the basis for future research. In this review, major advances and current debates are discussed, including MV receptor usage, the cellular basis of immunosuppression, the suspected role of MV in "nonviral" diseases such as multiple sclerosis and Paget's disease, and the controversy surrounding MV vaccine safety. PMID- 14527284 TI - The uncultured microbial majority. AB - Since the delineation of 12 bacterial phyla by comparative phylogenetic analyses of 16S ribosomal RNA in 1987 knowledge of microbial diversity has expanded dramatically owing to the sequencing of ribosomal RNA genes cloned from environmental DNA. Currently, only 26 of the approximately 52 identifiable major lineages, or phyla, within the domain Bacteria have cultivated representatives. Evidence from field studies indicates that many of the uncultivated phyla are found in diverse habitats, and some are extraordinarily abundant. In some important environments, including seawater, freshwater, and soil, many biologically and geochemically important organisms are at best only remotely related to any strain that has been characterized by phenotype or by genome sequencing. Genome sequence information that would allow ribosomal RNA gene trees to be related to broader patterns in microbial genome evolution is scant, and therefore microbial diversity remains largely unexplored territory. PMID- 14527285 TI - Pathways of oxidative damage. AB - The phenomenon of oxygen toxicity is universal, but only recently have we begun to understand its basis in molecular terms. Redox enzymes are notoriously nonspecific, transferring electrons to any good acceptor with which they make electronic contact. This poses a problem for aerobic organisms, since molecular oxygen is small enough to penetrate all but the most shielded active sites of redox enzymes. Adventitious electron transfers to oxygen create superoxide and hydrogen peroxide, which are partially reduced species that can oxidize biomolecules with which oxygen itself reacts poorly. This review attempts to present our still-incomplete understanding of how reactive oxygen species are formed inside cells and the mechanisms by which they damage specific target molecules. The vulnerability of cells to oxidation lies at the root of obligate anaerobiosis, spontaneous mutagenesis, and the use of oxidative stress as a biological weapon. PMID- 14527286 TI - Gene organization: selection, selfishness, and serendipity. AB - The apparati behind the replication, transcription, and translation of prokaryotic and eukaryotic genes are quite different. Yet in both classes of organisms, genes may be organized in their respective chromosomes in similar ways by virtue of similarly acting selective forces. In addition, some gene organizations reflect biology unique to each class of organisms. Levels of organization are more complex than those of the simple operon. Multiple transcription units may be organized into larger units, local control regions may act over large chromosomal regions in eukaryotic chromosomes, and cis-acting genes may control the expression of downstream genes in all classes of organisms. All these mechanisms lead to genomes being far more organized, in both prokaryotes and eukaryotes, than hitherto imagined. PMID- 14527287 TI - Multiple sigma subunits and the partitioning of bacterial transcription space. AB - Promoter recognition in eubacteria is carried out by the initiation factor sigma, which binds RNA polymerase and initiates transcription. Cells have one housekeeping factor and a variable number of alternative sigma factors that possess different promoter-recognition properties. The cell can choose from its repertoire of sigmas to alter its transcriptional program in response to stress. Recent structural information illuminates the process of initiation and also shows that the two key sigma domains are structurally conserved, even among diverse family members. We use the sigma repertoire of Escherichia coli, Bacillus subtilis, Streptomyces coelicolor, and cyanobacteria to illustrate the different strategies utilized to organize transcriptional space using multiple sigma factors. PMID- 14527288 TI - Natural selection and the emergence of a mutation phenotype: an update of the evolutionary synthesis considering mechanisms that affect genome variation. AB - Most descriptions of evolution assume that all mutations are completely random with respect to their potential effects on survival. However, much like other phenotypic variations that affect the survival of the descendants, intrinsic variations in the probability, type, and location of genetic change can feel the pressure of natural selection. From site-specific recombination to changes in polymerase fidelity and repair of DNA damage, an organism's gene products affect what genetic changes occur in its genome. Through the action of natural selection on these gene products, potentially favorable mutations can become more probable than random. With examples from variation in bacterial surface proteins to the vertebrate immune response, it is clear that a great deal of genetic change is better than "random" with respect to its potential effect on survival. Indeed, some potentially useful mutations are so probable that they can be viewed as being encoded implicitly in the genome. An updated evolutionary theory includes emergence, under selective pressure, of genomic information that affects the probability of different classes of mutation, with consequences for genome survival. PMID- 14527289 TI - Archeal DNA replication: eukaryal proteins in a bacterial context. AB - Genome sequences of a number of archaea have revealed an apparent paradox in the phylogenies of the bacteria, archaea, and eukarya, as well as an intriguing set of problems to be resolved in the study of DNA replication. The archaea, long thought to be bacteria, are not only different enough to merit their own domain but also appear to be an interesting mosaic of bacterial, eukaryal, and unique features. Most archaeal proteins participating in DNA replication are more similar in sequence to those found in eukarya than to analogous replication proteins in bacteria. However, archaea have only a subset of the eukaryal replication machinery, apparently needing fewer polypeptides and structurally simpler complexes. The archaeal replication apparatus also contains features not found in other organisms owing, in part, to the broad range of environmental conditions, some extreme, in which members of this domain thrive. In this review the current knowledge of the mechanisms governing DNA replication in archaea is summarized and the similarities and differences of those of bacteria and eukarya are highlighted. PMID- 14527290 TI - Molecular genetics of Mycobacterium tuberculosis pathogenesis. AB - Tuberculosis (TB) has afflicted humankind throughout history. Approximately one third of the world's population is currently infected with Mycobacterium tuberculosis and nearly two million people die of TB annually. Although much has been learned about the structure of the tubercle bacillus, the epidemiology of TB, the physiological and immunological responses of the host to infection, and the physiology of M. tuberculosis in laboratory broth cultures, much of the basic biology of M. tuberculosis in its natural setting (the infected human) remains to be elucidated. Within the past decade, there have been remarkable advances in the development of genetic and molecular biological tools with which to study M. tuberculosis. This review discusses the approaches that have been employed and the progress that has been made in discovering how M. tuberculosis has achieved its prowess as a successful pathogen. PMID- 14527291 TI - The bacterial RecA protein as a motor protein. AB - The bacterial RecA protein plays a central role in the repair of stalled replication forks, double-strand break repair, general recombination, induction of the SOS response, and SOS mutagenesis. The major activity of RecA in DNA metabolism is the promotion of DNA strand exchange reactions. RecA is the prototype for a ubiquitous family of proteins but exhibits a few activities that some of its eukaryotic, archaeal, and viral homologs appear to lack. In particular, the bacterial RecA protein possesses an apparent motor function that is not evident in the reactions promoted by the eukaryotic Rad51 protein. This motor may be needed only in a subset of the DNA metabolism contexts in which RecA protein functions. Models for the coupling of DNA strand exchange to ATP hydrolysis are examined. PMID- 14527292 TI - DNA mismatch repair: molecular mechanisms and biological function. AB - DNA mismatch repair (MMR) guards the integrity of the genome in virtually all cells. It contributes about 1000-fold to the overall fidelity of replication and targets mispaired bases that arise through replication errors, during homologous recombination, and as a result of DNA damage. Cells deficient in MMR have a mutator phenotype in which the rate of spontaneous mutation is greatly elevated, and they frequently exhibit microsatellite instability at mono- and dinucleotide repeats. The importance of MMR in mutation avoidance is highlighted by the finding that defects in MMR predispose individuals to hereditary nonpolyposis colorectal cancer. In addition to its role in postreplication repair, the MMR machinery serves to police homologous recombination events and acts as a barrier to genetic exchange between species. PMID- 14527294 TI - The secret lives of the pathogenic mycobacteria. AB - Pathogenic mycobacteria, including the causative agents of tuberculosis and leprosy, are responsible for considerable morbidity and mortality worldwide. A hallmark of these pathogens is their tendency to establish chronic infections that produce similar pathologies in a variety of hosts. During infection, mycobacteria reside in macrophages and induce the formation of granulomas, organized immune complexes of differentiated macrophages, lymphocytes, and other cells. This review summarizes our understanding of Mycobacterium-host cell interactions, the bacterial-granuloma interface, and mechanisms of bacterial virulence and persistence. In addition, we highlight current controversies and unanswered questions in these areas. PMID- 14527295 TI - Bacterial biofilms: an emerging link to disease pathogenesis. AB - The role of biofilms in the pathogenesis of some chronic human infections is now widely accepted. However, the criteria used to determine whether a given infection is caused by biofilms remain unclear. In this chapter we discuss three infections that are caused by biofilms--infectious kidney stones, bacterial endocarditis, and cystic fibrosis lung infections--and focus on the role of the biofilm in disease pathogenesis. Biofilms are also important as environmental reservoirs for pathogens, and the biofilm growth mode may provide organisms with survival advantages in natural environments and increase their virulence. The consequences of pathogens living in environmental biofilms and an analysis of some specific environmental biofilm systems are presented. PMID- 14527293 TI - Kaposi's sarcoma-associated herpesvirus immunoevasion and tumorigenesis: two sides of the same coin? AB - Kaposi's sarcoma-associated herpesvirus (KSHV) [or human herpesvirus 8 (HHV-8)] is the most frequent cause of malignancy among AIDS patients. KSHV and related herpesviruses have extensively pirated cellular cDNAs from the host genome, providing a unique opportunity to examine the range of viral mechanisms for controlling cell proliferation. Many of the viral regulatory homologs encode proteins that directly inhibit host adaptive and innate immunity. Other viral proteins target retinoblastoma protein and p53 control of tumor suppressor pathways, which also play key effector roles in intracellular immune responses. The immune evasion strategies employed by KSHV, by targeting tumor suppressor pathways activated during immune system signaling, may lead to inadvertent cell proliferation and tumorigenesis in susceptible hosts. PMID- 14527296 TI - Race, ancestry, and genes: implications for defining disease risk. AB - Geneticists are interested in finding genes associated with disease. Because of widespread health disparities, race is a variable that is often said to be relevant in this context. The idea is that members of a preconceived "race" share common ancestry that may include genetic risk factors. Human variation has been shaped by the long-term processes of population history, and population samples that reflect that history carry statistical information about shared genetic variation or "ancestry." But race is an elusive concept and a term difficult even to define rigorously. Unfortunately, these problems are neither new nor related to recent genetic knowledge. Race is also one of the most politically charged subjects in American life because its associated sociocultural component has notoriously led to categorical treatment that has been misleading and politically misused. There are ways in which the concept of race (whether or not the term is used) can be a legitimate tool in the search for disease-associated genes. But in that context race reflects deeply confounded cultural as well as biological factors, and a careful distinction must be made between race as a statistical risk factor and causal genetic variables. PMID- 14527297 TI - Gene annotation: prediction and testing. AB - Fifty years after the publication of DNA structure, the whole human genome sequence will be officially finished. This achievement marks the beginning of the task to catalogue every human gene and identify each of their function expression patterns. Currently, researchers estimate that there are about 30,000 human genes and approximately 70% of these can be automatically predicted using a combination of ab initio and similarity-based programs. However, to experimentally investigate every gene's function, the research community requires a high-quality annotation of alternative splicing, pseudogenes, and promoter regions that can only be provided by manual intervention. Manual curation of the human genome will be a long-term project as experimental data are continually produced to confirm or refine the predictions, and new features such as noncoding RNAs and enhancers have not been fully identified. Such a highly curated human gene-set made publicly available will be a great asset for the experimental community and for future comparative genome projects. PMID- 14527298 TI - The Drosophila melanogaster genome. AB - Drosophila's importance as a model organism made it an obvious choice to be among the first genomes sequenced, and the Release 1 sequence of the euchromatic portion of the genome was published in March 2000. This accomplishment demonstrated that a whole genome shotgun (WGS) strategy could produce a reliable metazoan genome sequence. Despite the attention to sequencing methods, the nucleotide sequence is just the starting point for genome-wide analyses; at a minimum, the genome sequence must be interpreted using expressed sequence tag (EST) and complementary DNA (cDNA) evidence and computational tools to identify genes and predict the structures of their RNA and protein products. The functions of these products and the manner in which their expression and activities are controlled must then be assessed-a much more challenging task with no clear endpoint that requires a wide variety of experimental and computational methods. We first review the current state of the Drosophila melanogaster genome sequence and its structural annotation and then briefly summarize some promising approaches that are being taken to achieve an initial functional annotation. PMID- 14527299 TI - Forensics and mitochondrial DNA: applications, debates, and foundations. AB - Debate on the validity and reliability of scientific methods often arises in the courtroom. When the government (i.e., the prosecution) is the proponent of evidence, the defense is obliged to challenge its admissibility. Regardless, those who seek to use DNA typing methodologies to analyze forensic biological evidence have a responsibility to understand the technology and its applications so a proper foundation(s) for its use can be laid. Mitochondrial DNA (mtDNA), an extranuclear genome, has certain features that make it desirable for forensics, namely, high copy number, lack of recombination, and matrilineal inheritance. mtDNA typing has become routine in forensic biology and is used to analyze old bones, teeth, hair shafts, and other biological samples where nuclear DNA content is low. To evaluate results obtained by sequencing the two hypervariable regions of the control region of the human mtDNA genome, one must consider the genetically related issues of nomenclature, reference population databases, heteroplasmy, paternal leakage, recombination, and, of course, interpretation of results. We describe the approaches, the impact some issues may have on interpretation of mtDNA analyses, and some issues raised in the courtroom. PMID- 14527300 TI - Creationism and intelligent design. AB - Creationism, the rejection of evolution in favor of supernatural design, comes in many varieties besides the common young-earth Genesis version. Creationist attacks on science education have been evolving in the last few years through the alliance of different varieties. Instead of calls to teach "creation science," one now finds lobbying for "intelligent design" (ID). Guided by the Discovery Institute's "Wedge strategy," the ID movement aims to overturn evolution and what it sees as a pernicious materialist worldview and to renew a theistic foundation to Western culture, in which human beings are recognized as being created in the image of God. Common ID arguments involving scientific naturalism, "irreducible complexity," "complex specified information," and "icons of evolution," have been thoroughly examined and refuted. Nevertheless, from Kansas to Ohio to the U.S. Congress, ID continues lobbying to teach the controversy, and scientists need to be ready to defend good evolution education. PMID- 14527301 TI - Peroxisome biogenesis disorders. AB - The peroxisome biogenesis disorders (PBDs) comprise 12 autosomal recessive complementation groups (CGs). The multisystem clinical phenotype varies widely in severity and results from disturbances in both development and metabolic homeostasis. Progress over the last several years has lead to identification of the genes responsible for all of these disorders and to a much improved understanding of the biogenesis and function of the peroxisome. Increasing availability of mouse models for these disorders offers hope for a better understanding of their pathophysiology and for development of therapies that might especially benefit patients at the milder end of the clinical phenotype. PMID- 14527302 TI - Sequence divergence, functional constraint, and selection in protein evolution. AB - The genome sequences of multiple species has enabled functional inferences from comparative genomics. A primary objective is to infer biological functions from the conservation of homologous DNA sequences between species. A second, more difficult, objective is to understand what functional DNA sequences have changed over time and are responsible for species' phenotypic differences. The neutral theory of molecular evolution provides a theoretical framework in which both objectives can be explicitly tested. Development of statistical tests within this framework has provided insight into the evolutionary forces that constrain and in some cases change DNA sequences and the resulting patterns that emerge. In this article, we review recent work on how functional constraint and changes in protein function are inferred from protein polymorphism and divergence data. We relate these studies to our understanding of the neutral theory and adaptive evolution. PMID- 14527303 TI - Molecular pathogenesis of pancreatic cancer. AB - Pancreatic cancer is an almost universally fatal disease, with a five-year survival rate of 5%. Research into both sporadic and inherited forms of pancreatic cancer has yielded tremendous advances in the understanding of this disease at the molecular level. Elucidating genetic alterations in pancreatic cancer has identified various abnormalities ranging from gross chromosomal abnormalities to point mutations, many of which influence the development and progression of pancreatic cancer. Identifying precursor lesions within pancreatic ducts has led to the formulation of a progression model of pancreatic cancer and subsequent identification of early- and late-stage changes leading to invasive cancer. Ultimately, understanding the genetic events underlying the development of pancreatic cancer may serve as a useful adjunct in the screening and treatment of patients suffering from, or at risk for, pancreatic cancer. PMID- 14527304 TI - The inherited basis of diabetes mellitus: implications for the genetic analysis of complex traits. AB - Diabetes encompasses a heterogeneous group of diseases, each with a substantial genetic component. We review the division of diabetes into different subtypes based on clinical phenotype, the fruitful pursuit of genes underlying monogenic forms of the disease, the successes and drawbacks of whole-genome linkage scans in type 1 and type 2 diabetes, and the recent identification of several diabetes genes by large association studies. We use the lessons learned from this extensive body of evidence to illustrate general implications for the genetic analysis of complex traits. PMID- 14527305 TI - Patterns of human genetic diversity: implications for human evolutionary history and disease. AB - Since the completion of the human genome sequencing project, the discovery and characterization of human genetic variation is a principal focus for future research. Comparative studies across ethnically diverse human populations and across human and nonhuman primate species is important for reconstructing human evolutionary history and for understanding the genetic basis of human disease. In this review, we summarize data on patterns of human genetic diversity and the evolutionary forces (mutation, genetic drift, migration, and selection) that have shaped these patterns of variation across both human populations and the genome. African population samples typically have higher levels of genetic diversity, a complex population substructure, and low levels of linkage disequilibrium (LD) relative to non-African populations. We discuss these differences and their implications for mapping disease genes and for understanding how population and genomic diversity have been important in the evolution, differentiation, and adaptation of humans. PMID- 14527306 TI - Human nonsyndromic sensorineural deafness. AB - Given the unique biological requirements of sound transduction and the selective advantage conferred upon a species capable of sensitive sound detection, it is not surprising that up to 1% of the approximately 30,000 or more human genes are necessary for hearing. There are hundreds of monogenic disorders for which hearing loss is one manifestation of a syndrome or the only disorder and therefore is nonsyndromic. Herein we review the supporting evidence for identifying over 30 genes for dominantly and recessively inherited, nonsyndromic, sensorineural deafness. The state of knowledge concerning their biological roles is discussed in the context of the controversies within an evolving understanding of the intricate molecular machinery of the inner ear. PMID- 14527307 TI - Enzyme therapy for lysosomal storage disease: principles, practice, and prospects. AB - Over the past three decades, enzyme therapy for lysosomal storage diseases has moved from an academic pursuit to direct delivery of effective clinical care for affected patients and families. This success is based on understanding the complexities of lysosomal biogenesis, lysosomal hydrolase sorting and hydrolytic requirements, and the target sites of pathology of these diseases. This article reviews these concepts and their application to the treatment of affected patients with Gaucher disease, Fabry disease, and mucopolysaccharidosis I. The principles, progress, and practice in these diseases provide prototypes for expansion of enzyme therapy to a growing set of these diseases. PMID- 14527308 TI - Nonsyndromic seizure disorders: epilepsy and the use of the internet to advance research. AB - The progress in understanding the genetics of nonsyndromic epilepsy is the direct result of dramatic advances made by the Human Genome Project. The development of thousands of precisely mapped genetic markers and the nearly complete sequencing of the entire human genome in 2001 allowed genetic researchers in epilepsy to identify many loci and genes as causal in inherited idiopathic epilepsy. This substantial increase in information has required the development of accurate and online bioinformatic databases. Only the Internet can enable such large amounts of precise DNA sequence information to be transferred to researchers. Along with the construction of these databases has been the development of efficient search algorithms for specific DNA sequences and genetic information. This article summarizes the effect that this burst of new genomic information has had on research aimed at discovering the underlying genetic factors for nonsyndromic epilepsy. Many of the web sites important to epilepsy gene discovery are listed and discussed in this article, including sites with extensive information on genetic markers, genetic analysis, gene sequence, gene expression, gene mutations, and DNA sequence variation. Continued acquisition of information on naturally occurring DNA sequence variants will greatly help research directed towards understanding the genetic susceptibility of the common, nonsyndromic epilepsies and will lead to the promise of personalized medicine. PMID- 14527309 TI - The genetics of narcolepsy. AB - Human narcolepsy is a genetically complex disorder. Family studies indicate a 20 40 times increased risk of narcolepsy in first-degree relatives and twin studies suggest that nongenetic factors also play a role. The tight association between narcolepsy-cataplexy and the HLA allele DQB1*0602 suggests that narcolepsy has an autoimmune etiology. In recent years, extensive genetic studies in animals, using positional cloning in dogs and gene knockouts in mice, have identified abnormalities in hypothalamic hypocretin (orexin) neurotransmission as key to narcolepsy pathophysiology. Though most patients with narcolepsy-cataplexy are hypocretin deficient, mutations or polymorphisms in hypocretin-related genes are extremely rare. It is anticipated that susceptibility genes that are independent of HLA and impinge on the hypocretin neurotransmitter system are isolated in human narcolepsy. PMID- 14527310 TI - Structural, functional, and molecular MR imaging of the microvasculature. AB - Magnetic resonance imaging (MRI) is widely applied for functional imaging of the microcirculation and for functional and structural studies of the microvasculature. The interest in the capabilities of MRI in noninvasively monitoring changes in vascular structure and function expanded over the past years, with specific efforts directed toward the development of novel imaging methods for quantification of angiogenesis. Molecular imaging approaches hold promise for further expansion of the ability to characterize the microvasculature. Exciting applications for MRI are emerging in the study of the biology of microvessels and in the evaluation of potential pharmaceutical modulators of vascular function and development, and preclinical MRI tools can serve for the design of mechanism-of-action-based noninvasive clinical methods for monitoring response to therapy. The aim of this review is to provide a current snapshot of recent developments in this rapidly evolving field. PMID- 14527311 TI - Temporal dynamics of brain anatomy. AB - The brain changes profoundly in structure and function during development and as a result of diseases such as the dementias, schizophrenia, multiple sclerosis, and tumor growth. Strategies to measure, map, and visualize these brain changes are of immense value in basic and clinical neuroscience. Algorithms that map brain change with sufficient spatial and temporal sensitivity can also assess drugs that aim to decelerate or arrest these changes. In neuroscience studies, these tools can reveal subtle brain changes in adolescence and old age and link these changes with measurable differences in brain function and cognition. Early detection of brain change in patients at risk for dementia; tumor recurrence; or relapsing-remitting conditions, such as multiple sclerosis, is also vital for optimizing therapy. We review a variety of mathematical and computational approaches to detect structural brain change with unprecedented sensitivity, both spatially and temporally. The resulting four-dimensional (4-D) maps of brain anatomy are warehoused in population-based brain atlases. Here, statistical tools compare brain changes across subjects and across populations, adjusting for complex differences in brain structure. Brain changes in an individual can be compared with a normative database comprised of subjects matched for age, gender, and other demographic factors. These dynamic brain maps offer key biological markers for understanding disease progression and testing therapeutic response. The early detection of disease-related brain changes is also critical for possible pre-emptive intervention before the ravages of disease have set in. PMID- 14527312 TI - Modeling total heart function. AB - Computational models of the electrical and mechanical function of the heart are reviewed. These models attempt to explain the integrated function of the heart in terms of ventricular anatomy, the structure and material properties of myocardial tissue, the membrane ion channels, and calcium handling and myofilament mechanics of cardiac myocytes. The models have established the computational framework for linking the structure and function of cardiac cells and tissue to the integrated behavior of the intact heart, but many more aspects of physiological function, including metabolic and signal transduction pathways, need to be included before significant progress can be made in understanding many disease processes. PMID- 14527313 TI - The engineering of gene regulatory networks. AB - The rapid accumulation of genetic information and advancement of experimental techniques have opened a new frontier in biomedical engineering. With the availability of well-characterized components from natural gene networks, the stage has been set for the engineering of artificial gene regulatory networks with sophisticated computational and functional capabilities. In these efforts, the ability to construct, analyze, and interpret qualitative and quantitative models is becoming increasingly important. In this review, we consider the current state of gene network engineering from a combined experimental and modeling perspective. We discuss how networks with increased complexity are being constructed from simple modular components and how quantitative deterministic and stochastic modeling of these modules may provide the foundation for accurate in silico representations of gene regulatory network function in vivo. PMID- 14527314 TI - Engineered nanomaterials for biophotonics applications: improving sensing, imaging, and therapeutics. AB - Advances in chemistry and physics are providing an expanding array of nanostructured materials with unique and powerful optical properties. These nanomaterials provide a new set of tools that are available to biomedical engineers, biologists, and medical scientists who seek new tools as biosensors and probes of biological fluids, cells, and tissue chemistry and function. Nanomaterials are also being used to develop optically controlled devices for applications such as modulated drug delivery as well as optical therapeutics. This review discusses applications that have been successfully demonstrated using nanomaterials including semiconductor nanocrystals, gold nanoparticles, gold nanoshells, and silver plasmon resonant particles. PMID- 14527315 TI - Neural tissue engineering: strategies for repair and regeneration. AB - Nerve regeneration is a complex biological phenomenon. In the peripheral nervous system, nerves can regenerate on their own if injuries are small. Larger injuries must be surgically treated, typically with nerve grafts harvested from elsewhere in the body. Spinal cord injury is more complicated, as there are factors in the body that inhibit repair. Unfortunately, a solution to completely repair spinal cord injury has not been found. Thus, bioengineering strategies for the peripheral nervous system are focused on alternatives to the nerve graft, whereas efforts for spinal cord injury are focused on creating a permissive environment for regeneration. Fortunately, recent advances in neuroscience, cell culture, genetic techniques, and biomaterials provide optimism for new treatments for nerve injuries. This article reviews the nervous system physiology, the factors that are critical for nerve repair, and the current approaches that are being explored to aid peripheral nerve regeneration and spinal cord repair. PMID- 14527316 TI - Metabolic engineering: advances in modeling and intervention in health and disease. AB - The field of metabolic engineering encompasses a powerful set of tools that can be divided into (a) methods to model complex metabolic pathways and (b) techniques to manipulate these pathways for a desired metabolic outcome. These tools have recently seen increased utility in the medical arena, and this paper aims to review significant accomplishments made using these approaches. The modeling of metabolic pathways has been applied to better understand disease state physiology in a variety of cellar, subcellular, and organ systems, including the liver, heart, mitochondria, and cancerous cells. Metabolic pathway engineering has been used to generate cells with novel biochemical functions for therapeutic use, and specific examples are provided in the areas of glycosylation engineering and dopamine-replacement therapy. In order to document the potential of applying both metabolic modeling and pathway manipulation, we describe pertinent advances in the field of diabetes research. Undoubtedly, as the field of metabolic engineering matures and is applied to a wider array of problems, new advances and therapeutic strategies will follow. PMID- 14527317 TI - Biomonitoring with wireless communications. AB - Wireless biomonitoring, first used in human beings for fetal heart-rate monitoring more than 30 years ago, has now become a technology for remote sensing of patients' activity, blood pulse pressure, oxygen saturation, internal pressures, orthopedic device loading, and gastrointestinal endoscopy. Technical advances in miniaturization and wireless communications have enabled development of monitoring devices that can be made available for general use by individuals/patients and caregivers. New methods for short-range wireless communications not encumbered by radio spectrum restrictions (e.g., ultra wideband) will enable applications of wireless monitoring without interference in ambulatory subjects, in home care, and in hospitals. PMID- 14527319 TI - Time-reversal acoustics in biomedical engineering. AB - Time reversal is a very powerful method for focusing through complex and heterogeneous media and shows very promising results in biomedical applications. In this paper, we review some of the main applications investigated during the past decade. An iterative implementation of the time-reversal process allows tracking gallstones in real time during lithotripsy treatments. In this application domain, a smart exploitation of the reverberations in solid waveguides permits the focusing of high-amplitude ultrasonic shock waves with a small number of transducers. Finally, because time reversal is able to correct the strong distortions induced by the skull bone on ultrasonic propagation, this adaptive focusing technique is very promising for ultrasonic hyperthermia brain therapy. PMID- 14527318 TI - The tissue engineeting puzzle: a molecular perspective. AB - The inability of biomaterial scaffolds to functionally integrate into surrounding tissue is one of the major roadblocks to developing new biomaterials and tissue engineering scaffolds. Despite considerable advances, current approaches to engineering cell-surface interactions fall short in mimicking the complexity of signals through which surrounding tissue regulates cell behavior. Cells adhere and interact with their extracellular environment via integrins, and their ability to activate associated downstream signaling pathways depends on the character of adhesion complexes formed between cells and their extracellular matrix. In particular, alpha5beta1 and alphavbeta3 integrins are central to regulating downstream events, including cell survival and cell-cycle progression. In contrast to previous findings that alphavbeta3 integrins promote angiogenesis, recent evidence argues that alphavbeta3 integrins may act as negative regulators of proangiogenic integrins such as alpha5beta1. This suggests that fibronectin is critical for scaffold vascularization because it is the only mammalian adhesion protein that binds and activates alpha5beta1 integrins. Cells are furthermore capable of stretching fibronectin matrices such that the protein partially unfolds, and recent computational simulations provide structural models of how mechanical stretching affects fibronectin function. We propose a model whereby excessive tension generated by cells in contact to biomaterials may in fact render fibronectin fibrils nonangiogenic and potentially inhibit vascularization. The model could explain why current biomaterials independent of their surface chemistries and textures fail to vascularize. PMID- 14527320 TI - With the help of giants. AB - A childhood fascination with animals, plants, and insects was aided and abetted by many giants, beginning with my parents. The Bronx High School of Science and the City College of New York (CCNY) made a solid and priceless grounding in chemistry and biology available free of charge. Abe Mazur at CCNY revealed the wonders of biochemistry and illustrated that it was possible to pursue these wonders while being paid to do so. He also directed me to Duke University Medical School for PhD work under the tutelage of Phil Handler. With the exception of a sabbatical year at Harvard with Frank Westheimer, my entire career has been spent at Duke serving under three fine and supportive chairmen: Handler, Hill, and Raetz. The premier discoveries to emanate from my laboratory have been the sulfite oxidase, the several superoxide dismutases, the manganese catalase, and the catalase/peroxidase. Many other topics piqued my interest and resulted in ~ 400 publications. Herein I have recounted some of the circumstances surrounding that work and named a few of the people involved. The first 20 years I worked happily at the bench and the next 35 years just as happily facilitating the work of younger people. It has been so rewarding that I wish for nothing more than to be allowed to keep at it. PMID- 14527321 TI - Function and structure of complex II of the respiratory chain. AB - Complex II is the only membrane-bound component of the Krebs cycle and in addition functions as a member of the electron transport chain in mitochondria and in many bacteria. A recent X-ray structural solution of members of the complex II family of proteins has provided important insights into their function. One feature of the complex II structures is a linear electron transport chain that extends from the flavin and iron-sulfur redox cofactors in the membrane extrinsic domain to the quinone and b heme cofactors in the membrane domain. Exciting recent developments in relation to disease in humans and the formation of reactive oxygen species by complex II point to its overall importance in cellular physiology. PMID- 14527322 TI - Protein-lipid interplay in fusion and fission of biological membranes. AB - Disparate biological processes involve fusion of two membranes into one and fission of one membrane into two. To formulate the possible job description for the proteins that mediate remodeling of biological membranes, we analyze the energy price of disruption and bending of membrane lipid bilayers at the different stages of bilayer fusion. The phenomenology and the pathways of the well-characterized reactions of biological remodeling, such as fusion mediated by influenza hemagglutinin, are compared with those studied for protein-free bilayers. We briefly consider some proteins involved in fusion and fission, and the dependence of remodeling on the lipid composition of the membranes. The specific hypothetical mechanisms by which the proteins can lower the energy price of the bilayer rearrangement are discussed in light of the experimental data and the requirements imposed by the elastic properties of the bilayer. PMID- 14527323 TI - The many faces of vitamin B12: catalysis by cobalamin-dependent enzymes. AB - Vitamin B12 is a complex organometallic cofactor associated with three subfamilies of enzymes: the adenosylcobalamin-dependent isomerases, the methylcobalamin-dependent methyltransferases, and the dehalogenases. Different chemical aspects of the cofactor are exploited during catalysis by the isomerases and the methyltransferases. Thus, the cobalt-carbon bond ruptures homolytically in the isomerases, whereas it is cleaved heterolytically in the methyltransferases. The reaction mechanism of the dehalogenases, the most recently discovered class of B12 enzymes, is poorly understood. Over the past decade our understanding of the reaction mechanisms of B12 enzymes has been greatly enhanced by the availability of large amounts of enzyme that have afforded detailed structure-function studies, and these recent advances are the subject of this review. PMID- 14527324 TI - Covalent trapping of protein-DNA complexes. AB - High-resolution structural studies of protein-DNA complexes have proven to be an invaluable means of understanding the diverse functions of proteins that manage the genome. Most of the structures determined to date represent proteins bound noncovalently to various DNA sequences or structures. Although noncovalent complexation is often adequate to study the structures of proteins that have robust, specific interactions with DNA, it is poorly suited to the study of transient intermediates in enzyme-catalyzed DNA processing reactions or of complexes that exist in multiple equilibrating forms. In recent years, strategies developed for the covalent trapping of protein-DNA complexes have begun to show promise as a window into an otherwise inaccessible world of structure. PMID- 14527325 TI - The dynamics of chromosome organization and gene regulation. AB - With the sequence of the human genome now complete, studies must focus on how the genome is functionally organized within the confines of the cell nucleus and the dynamic interplay between the genome and its regulatory factors to effectively control gene expression and silencing. In this review I describe our current state of knowledge with regard to the organization of chromosomes within the nucleus and the positioning of active versus inactive genes. In addition, I discuss studies on the dynamics of chromosomes and specific genetic loci within living cells and its relationship to gene activity and the cell cycle. Furthermore, our current understanding of the distribution and dynamics of RNA polymerase II transcription factors is discussed in relation to chromosomal loci and other nuclear domains. PMID- 14527326 TI - Dynamics of cell surface molecules during T cell recognition. AB - Recognition of foreign antigens by T lymphocytes is a very important component of vertebrate immunity-vital to the clearance of pathogenic organisms and particular viruses and necessary, indirectly, for the production of high affinity antibodies. T cell recognition is mediated by the systematic scanning of cell surfaces by T cells, which collectively express many antigen receptors. When the appropriate antigenic peptide bound to a molecule of the major histocompatibility complex is found-even in minute quantities-a series of elaborate cell-surface molecule and internal rearrangements take place. The sequence of events and the development of techniques required to observe these events have significantly enhanced our understanding of T cell recognition and may find application in other systems of transient cell:cell interactions as well. PMID- 14527327 TI - Proteomics. AB - Fueled by ever-growing DNA sequence information, proteomics-the large scale analysis of proteins-has become one of the most important disciplines for characterizing gene function, for building functional linkages between protein molecules, and for providing insight into the mechanisms of biological processes in a high-throughput mode. It is now possible to examine the expression of more than 1000 proteins using mass spectrometry technology coupled with various separation methods. High-throughput yeast two-hybrid approaches and analysis of protein complexes using affinity tag purification have yielded valuable protein protein interaction maps. Large-scale protein tagging and subcellular localization projects have provided considerable information about protein function. Finally, recent developments in protein microarray technology provide a versatile tool to study protein-protein, protein-nucleic acid, protein-lipid, enzyme-substrate, and protein-drug interactions. Other types of microarrays, though not fully developed, also show great potential in diagnostics, protein profiling, and drug identification and validation. This review discusses high throughput technologies for proteome analysis and their applications. Also discussed are the approaches used for the integrated analysis of the voluminous sets of data generated by proteome analysis conducted on a global scale. PMID- 14527328 TI - The structural basis of large ribosomal subunit function. AB - The ribosome crystal structures published in the past two years have revolutionized our understanding of ribonucleoprotein structure, and more specifically, the structural basis of the peptide bonding forming activity of the ribosome. This review concentrates on the crystallographic developments that made it possible to solve these structures. It also discusses the information obtained from these structures about the three-dimensional architecture of the large ribosomal subunit, the mechanism by which it facilitates peptide bond formation, and the way antibiotics inhibit large subunit function. The work reviewed, taken as a whole, proves beyond doubt that the ribosome is an RNA enzyme, as had long been surmised on the basis of less conclusive evidence. PMID- 14527329 TI - Molecular basis of Pto-mediated resistance to bacterial speck disease in tomato. AB - The Pto gene in tomato confers gene-for-gene resistance to Pseudomonas syringae pv. tomato, the causative agent of bacterial speck disease. Pto was first introgressed from a wild species of tomato into cultivated tomato varieties over 60 years ago and is now widely used to control speck disease. Cloning of the Pto gene revealed that it encodes a cytoplasmically localized serine-threonine protein kinase. The molecular basis of gene-for-gene recognition in this pathosystem is the direct physical interaction of the Pto kinase with either of two Pseudomonas effector proteins, AvrPto and AvrPtoB. Upon recognition of AvrPto or AvrPtoB, the Pto kinase acts in concert with Prf, a leucine-rich repeat containing protein, to activate multiple signal transduction pathways. There has been much progress in understanding the evolutionary origin of the Pto gene, structural details about how the Pto kinase interacts with AvrPto and AvrPtoB, signaling steps downstream of Pto, and defense responses activated by the Pto pathway. Future work on this model system will focus on how the interaction of the Pto kinase with bacterial effector proteins activates signal transduction, defining the specific role of signaling components, and ultimately, determining which host defense responses are most responsible for inhibiting growth of the pathogen and suppressing symptoms of bacterial speck disease. PMID- 14527330 TI - Parasitic nematode interactions with mammals and plants. AB - Parasitic nematodes that infect humans, animals, and plants cause serious diseases that are deleterious to human health and agricultural productivity. Chemical and biological control methods have reduced the impact of these parasites. However, surviving environmental stages lead to persistent reinfection of host species. In addition, development of resistance to nematicides and anthelmintics by these parasites and reduced availability of some nematicides, for environmental protection, pose significant obstacles for current and future prospects of effective parasite control. Due to marked differences in host species, research on animal and plant parasitic nematodes often proceeds independently. Despite the differences between animals and plants, basic cellular properties are shared among these host organisms. Some common properties may be important for mechanisms [homologous or convergent (homoplastic)] by which nematodes successfully infect these diverse hosts or by which animal and plant hosts resist infections by these pathogens. Here we compare host/parasite interactions between plant parasitic nematodes (PPN) and animal parasitic nematodes, with an emphasis on mammalian hosts (MPN). Similarities and differences are considered in the context of progress on molecular dissection of these interactions. A comprehensive coverage is not possible in the space allotted. Instead, an illustrative approach is used to establish examples that, it is hoped, exemplify the value of the comparative approach. PMID- 14527331 TI - Advances in molecular-based diagnostics in meeting crop biosecurity and phytosanitary issues. AB - Awareness of crop biosecurity and phytosanitation has been heightened since 9/11 and the unresolved anthrax releases in October 2001. Crops are highly vulnerable to accidental or deliberate introductions of crop pathogens from outside U.S. borders. Strategic thinking about protection against deliberate or accidental release of a plant pathogen is an urgent priority. Rapid detection will be the key to success. This review summarizes recent progress in the development of rapid real-time PCR protocols and evaluates their effectiveness in a proposed nationwide network of diagnostic laboratories that will facilitate rapid diagnostics and improved communication. PMID- 14527332 TI - Development of alternative strategies for management of soilborne pathogens currently controlled with methyl bromide. AB - The current standard treatment for management of soilborne pests in some high value crop production systems is preplant fumigation with mixtures of methyl bromide and chloropicrin. With the impending phase-out of methyl bromide, the agricultural industries that rely on soil fumigation face the need for development of alternative pest management strategies. To maintain farm productivity, immediate term research has focused on evaluation of alternative fumigants, modification of current crop production practices to accommodate their use, and improvement of application technologies to reduce the environmental effects of fumigant applications. Longer-term research goals have focused on developing a more integrated approach for pest management that incorporates the use of cultural practices to reduce pathogen pressure, host resistance to disease, and biological approaches for stimulating plant growth and control of root diseases. PMID- 14527333 TI - Patterns of pesticide use in California and the implications for strategies for reduction of pesticides. AB - We used the California Pesticide Use Reports to study use of fungicides, bactericides, fumigants, and selected insecticides, primarily for vegetable, fruit, and nut production in California from 1993 to 2000. There were no obvious trends in decreased use of most compounds used to treat plant disease. However, growers have rapidly adopted recently introduced "conventional" compounds. There is very limited use of microbial biocontrol agents to control plant disease and no indication of an increase. We used case studies to explore the potential of different strategies to reduce pesticide use or risk. There have been reductions in use of organophosphate insecticides, largely by substitution with pyrethroids. Theoretically, replacement of "calendar spray" pesticide programs with "environmentally driven" programs could reduce pesticide use in years with lower disease pressure, but this assumes that the majority of growers currently use a "calendar spray" program and that growers who use less than recommended by an environmentally driven program would not increase their use. PMID- 14527334 TI - Ecology and epidemiology of benyviruses and plasmodiophorid vectors. AB - Beet necrotic yellow vein virus (BNYVV) and Beet soilborne mosaic virus (BSBMV) are members of the genus Benyvirus, and Burdock mottle virus (BdMV) is a tentative member. BNYVV and BSBMV are vectored by the plasmodiophorid Polymyxa betae, which has a worldwide distribution. Polymyxa betae is morphologically indistinguishable from P. graminis, but recent molecular studies support separation of the two species. The geographic distribution of BNYVV is also worldwide, but BSBMV has been identified only in the United States. In Europe and Japan, several genotypic strains of BNYVV have been identified, and those with a fifth RNA appear to be more aggressive. No thorough survey of genotypic variability of BNYVV or BSBMV has been conducted in the United States. However, both viruses are widespread and frequently found in the same field, infecting the same beet plant. The implications of this close proximity, with regard to disease incidence and severity, and for recombination, are uncertain. Recent technological advances that permit improved detection and quantification of these viruses and their vector offer tremendous research opportunities. PMID- 14527335 TI - Establishment of biotrophy by parasitic fungi and reprogramming of host cells for disease resistance. AB - Parasitic biotrophs such as mildews and rusts evolved specific mechanisms that keep host cells alive during infection. These fungi appear to absorb nutrients mainly by proton-symport-driven transporter proteins that reside in specialized feeding structures. Accumulating evidence suggests that biotrophic fungi both suppress induction of plant defense responses in physical proximity to infection sites and induce specific host genes for the establishment of biotrophy. The peculiarities of biotrophic pathogenesis likely reflect diverse types of plant disease-resistance responses. The cloning of race-specific resistance genes to powdery mildew infection and of genes required for their function provides first insights into molecular mechanisms enabling the host to recognize mildew effector components and suggests candidate mechanisms of resistance signaling. Resistance to powdery mildew fungi that result from mutations in host genes promises to shed light on mechanisms that are required for the establishment of disease susceptibility. PMID- 14527336 TI - Nutrition in the perioperative patient. AB - The association of malnutrition with surgical morbidity and mortality is well recognized. The question of whether this relationship is causal or simply an association in sick patients has been hotly debated. The field of nutrition support has grown out of the belief that correcting malnutrition will modify associated risks for poor outcome. It has been easier to substantiate this belief in some clinical situations than in others. The evidence for nutrition support during the perioperative period is reviewed and recommendations are made about where nutrition support is most useful and where it may be counterproductive. Some of the important unanswered questions about perioperative nutrition support are raised. PMID- 14527337 TI - Protein tyrosine phosphatase 1B participates in the down-regulation of erythropoietin receptor signalling. AB - Erythropoietin (EPO) is the principal hormone regulating the proliferation of erythroid precursors and their differentiation into erythrocytes. Binding of ligand to the cell-surface EPO-R (EPO receptor) induces dimerization and JAK2 (Janus kinase 2)-mediated tyrosine phosphorylation of the receptor. Less than 1% of the EPO-Rs are displayed on the cell surface; most of the receptor molecules are retained in intracellular compartments, including the ER (endoplasmic reticulum). Using pervanadate (PV) as a potent tool to inhibit cellular PTPs (protein tyrosine phosphatases), we demonstrated previously the accumulation of mature (endoglycosidase H-resistant) tyrosine-phosphorylated EPO-R [Cohen, Altaratz, Zick, Klingmuller and Neumann (1997) Biochem. J. 327, 391-397]. In the present study, we investigated the participation of the ER-associated PTP1B in the dephosphorylation of intracellular EPO-R. We demonstrate tyrosine phosphorylation of EPO-R in BOSC-23T cells co-expressing EPO-R and the 'substrate trapping' mutant form of PTP1B, PTP1B D181A (referred to as PTP1BD). In vivo interaction between EPO-R and PTP1B suggested that PTP1B dephosphorylates the EPO R intracellularly. Endoglycosidase H resistance of tyrosine-phosphorylated EPO-R in cells expressing PTP1BD suggested that mature EPO-R is dephosphorylated by PTP1B. Stimulation with EPO of cells co-expressing EPO-R and either PTP1BD or PTP1B resulted in an increase or decrease respectively in phosphotyrosine EPO-R. We thus suggest that PTP1B dephosphorylates EPO-stimulated EPO-R and participates in the down-regulation cascade of EPO-mediated signal transduction. PMID- 14527338 TI - Differential sorting of tyrosine kinases and phosphotyrosine phosphatases acting on band 3 during vesiculation of human erythrocytes. AB - One of the most intensively studied post-translational modifications of erythrocyte proteins is the phosphorylation of tyrosine residues of band 3, which is strictly regulated in vivo by PTKs (protein-tyrosine kinases) and PTPs (protein-phosphotyrosine phosphatases). Two PTKs (p72(syk) and p56/53(lyn)) and two PTP activities (PTP1B and SHPTP-2) have been immunologically identified so far in mature human erythrocytes. We have shown previously that band 3 undergoes tyrosine phosphorylation upon a decrease in cell volume, as occurs when erythrocytes treated with Ca(2+)/Ca(2+) ionophore (A23187) lose KCl and release microvesicles. Similar levels of band 3 tyrosine phosphorylation in vesicles and in the parent cells are induced by this treatment. However, we have found that tyrosine phosphorylation of band 3 in vesicles is more stable than in whole erythrocytes. Examination of how the identified PTPs and PTKs are partitioned between the vesicles and the remnant cells during vesiculation reveals that PTP1B, unlike the PTKs, is retained entirely in the parent cell compartment. Since a tight association between PTP1B and band 3 has been documented previously, we have investigated the partitioning of PTP1B and band 3 between the membrane and the membrane-skeletal fractions prepared from resting or Ca(2+)/A23187-treated cells. Our results rule out the possibility that the preferential retention of PTP1B within the cell was due to an increase in the amount of membrane-skeleton-associated band 3 (and of PTP1B) during the release of spectrin-free vesicles, suggesting a more complex modality of interaction of PTP1B with band 3 in the erythrocyte membrane. Analysis of erythrocytes of different cell ages revealed that PTP1B, unlike the other enzymes examined, was quantitatively conserved during erythrocyte aging. This suggests important roles for the down-regulation of tyrosine phosphorylation of band 3 in erythrocyte physiology, and for vesiculation as a mechanism of human erythrocyte senescence. PMID- 14527339 TI - Interleukin-8 binds to syndecan-2 on human endothelial cells. AB - Application of reverse transcription-PCR to total RNA prepared from TNF-alpha (tumour necrosis factor-alpha)-stimulated HUVECs (human umbilical vein endothelial cells) revealed that the syndecan-2 mRNA was up-regulated by this inflammatory stimulus. By immunoprecipitation using an anti-syndecan-2 antibody on TNF-alpha-stimulated HUVEC lysates, inflammation-induced interleukin-8 was found to be an interaction partner of this HS (heparan sulphate) proteoglycan, but not of any other syndecan on these cells. The glycosylated [Syn2(ect)(+HS)] and non-glycosylated [Syn2(ect)(-HS)] forms of Syn2(ect) (the syndecan-2 ectodomain) were purified from a stably transfected human cell line and from a bacterial expression system respectively. By CD spectroscopy, Syn2(ect) was found to adopt an all-beta secondary structure. The dissociation constant of Syn2(ect)(+HS) with respect to interleukin-8 binding was determined by isothermal fluorescence titrations to be 23 nM. Despite its lack of HS chains, Syn2(ect)( HS) exhibited significant binding to the chemokine, with a K (d) of >1 microM. Thus, in addition to glycosaminoglycan binding, protein-protein contacts might also contribute to the chemokine-proteoglycan interaction. PMID- 14527340 TI - Involvement of fatty acid pathways and cortical interaction of the pronuclear complex in Caenorhabditis elegans embryonic polarity. AB - BACKGROUND: Cell polarity is essential for many decisions made during development. While investigation of polarity-specific factors has yielded great insights into the polarization process, little is known on how these polarity specific factors link to the basic cellular mechanisms that function in non polarity aspects of the cell. To better understand the mechanisms that establish embryonic polarity, we investigated genes required for polarity in the one-cell C. elegans embryo that are also required for other non-polarity functions. This has led to the identification of the Pod-class of mutants that are characterized by osmosensitive embryos and defects in anterior-posterior polarity. RESULTS: Mutation in either of two loci of this class, emb-8 and pod-2, disrupts embryonic polarization and results in osmotically-sensitive embryos. Loss of emb-8, a previously uncharacterized polarity gene, causes mislocalization of PAR-3 and PAR 2 that molecularly mark the anterior and posterior cortices. emb-8 encodes NADPH cytochrome P450 reductase, a protein supplying electrons to cytochrome P450 family enzymes, some of which catalyze fatty acid modifications. Cloning of the previously characterized polarity gene pod-2 reveals it encodes acetyl-CoA carboxylase, an enzyme that catalyzes the first step in de novo fatty acid synthesis. Depletion of fatty acid synthase, the next enzyme in the biosynthetic pathway, by RNA-interference (RNAi) also causes similar loss of one-cell polarity. Furthermore, pod-2 polarity defects can be rescued by addition of exogenous fatty acids. By following the behavior of the pronucleus in emb-8 and pod-2 mutant embryos, we demonstrate that loss of polarity correlates with impaired interaction between the pronucleus-centrosome complex and the posterior cortex. CONCLUSIONS: The characterization of emb-8 and pod-2 mutant embryos suggests that the pronucleus-centrosome complex interaction with the cortex plays a direct role in establishing polarity and that fatty acid pathways are important for this polarizing event. PMID- 14527341 TI - A strategy for detection of known and unknown SNP using a minimum number of oligonucleotides applicable in the clinical settings. AB - Detection of unknown single nucleotide polymorphism (SNP) relies on large scale sequencing expeditions of genomic fragments or complex high-throughput chip technology. We describe a simplified strategy for fluorimetric detection of known and unknown SNP by proportional hybridization to oligonucleotide arrays based on optimization of the established principle of signal loss or gain that requires a drastically reduced number of matched or mismatched probes. The array consists of two sets of 18-mer oligonucleotide probes. One set includes overlapping oligos with 4-nucleotide tiling representing an arbitrarily selected "consensus" sequence (consensus-oligos), the other includes oligos specific for known SNP within the same genomic region (variant-oligos). Fluorescence-labeled DNA amplified from a homozygous source identical to the consensus represents the reference target and is co-hybridized with a differentially-labeled test sample. Lack of hybridization of the test sample to consensus- with simultaneous hybridization to variant-oligos designates a known allele. Lack of hybridization to consensus- and variant-oligos indicates a new allele. Detection of unknown variants in heterozygous samples depends upon fluorimetric analysis of signal intensity based on the principle that homozygous samples generate twice the amount of signal. This method can identify unknown SNP in heterozygous conditions with a sensitivity of 82% and specificity of 90%. This strategy should dramatically increase the efficiency of SNP detection throughout the human genome and will decrease the cost and complexity of applying genomic wide analysis in the context of clinical trials. PMID- 14527342 TI - Cross-reactivity between HLA-A2-restricted FLU-M1:58-66 and HIV p17 GAG:77-85 epitopes in HIV-infected and uninfected individuals. AB - BACKGROUND: The matrix protein of the influenza A virus and the matrix and capsid proteins of the human immunodeficiency virus (HIV) share striking structural similarities which may have evolutionary and biological significance. These similarities led us to hypothesize the existence of cross-reactivity between HLA A2-restricted FLU-M1:58-66 and HIV-1 p17 GAG:77-85 epitopes. METHODS: The hypothesis that these two epitopes are cross-reactive was tested by determining the presence and extent of FLU/GAG immune cross-reactivity in lymphocytes from HIV-seropositive and seronegative HLA-A2+ donors by cytotoxicity assays and tetramer analyses. Moreover, the molecular basis for FLU/GAG cross-reactivity in HIV-seropositive and seronegative donors was studied by comparing lymphocyte derived cDNA sequences corresponding to the TCR-beta variable regions, in order to determine whether stimulation of lymphocytes with either peptide results in the expansion of identical T-cell clonotypes. RESULTS: Here, we report evidence of cross-reactivity between FLU-M1:58-66 and HIV-1 p17 GAG:77-85 epitopes following in vitro stimulation of PBMC derived from either HIV-seropositive or seronegative HLA-A2+ donors as determined by cytotoxicity assays, tetramer analyses, and molecular clonotyping. CONCLUSION: These results suggest that immunity to the matrix protein of the influenza virus may drive a specific immune response to an HLA-A2-restricted HIV gag epitope in HIV-infected and uninfected donors vaccinated against influenza. PMID- 14527343 TI - Integration of Translational Research in the European Organization for Research and Treatment of Cancer Research (EORTC) Clinical Trial Cooperative Group Mechanisms. AB - The landscape for cancer research is profoundly different today from that only one decade ago. Basic science is moving rapidly and biotechnological revolutions in molecular targeting and immunology have completely modified the opportunities and concepts for cancer treatment. In contrast to the recent past where cytotoxic molecules were screened in the laboratory and then tested in early clinical studies with toxicity as endpoint instead of the often poorly defined mechanism for its potential anti-tumor effect, we now have entered the age of molecular therapeutics, rationally designed to target "strategic" checkpoints that underlie the malignant phenotype.Translational research in early clinical trials (Phase I and II) is an integral aspect of the development of the new generation of cancer drugs as it is necessary to implement radically different early phase clinical trial design and to validate new biological end-points if the full potential of these new agents is to be realized. The "proof of principle with mechanistic analysis" strategy will allow optimisation of therapy from the beginning, and provide important feedback to pre-clinical drug developers. Translational research is also essential in late (phase III) clinical trials in defining different patient populations that may benefit to differing degrees from new treatments, and thus provide further insight and refine clinical practice in a more and more patient-tailored approach. In this editorial we will discuss the integration of Translational Research in the Organization for Research and Treatment of Cancer (EORTC). PMID- 14527344 TI - Translational Medicine: A two-way road. AB - The purpose of translational research is to test, in humans, novel therapeutic strategies developed through experimentation. Translational research should be regarded as a two-way road: Bench to Bedside and Bedside to Bench. However, Bedside to Bench efforts have regrettably been limited because the scientific aspects are poorly understood by full time clinicians and the difficulty of dealing with humans poorly appreciated by basic scientists. Translational research would be most useful to the scientific community at large if journals would foster specific interest for the publication of ex vivo human observation. The review process for such work should be assigned to clinical scientists competent not only in the intricacies of molecular or cell biology but also intimate with the reality of Internal Review Boards, ethics committees, Governmental Regulatory Agencies and most importantly the humane aspects of dealing with sick individuals and their families. This approach may focus both basic and clinical scientists and those struggling to fill the gap between them on the effective treatment of diseases affecting women, men and children making translational research more than an interesting concept. PMID- 14527345 TI - A functional genomic analysis of cell morphology using RNA interference. AB - BACKGROUND: The diversity of metazoan cell shapes is influenced by the dynamic cytoskeletal network. With the advent of RNA-interference (RNAi) technology, it is now possible to screen systematically for genes controlling specific cell biological processes, including those required to generate distinct morphologies. RESULTS: We adapted existing RNAi technology in Drosophila cell culture for use in high-throughput screens to enable a comprehensive genetic dissection of cell morphogenesis. To identify genes responsible for the characteristic shape of two morphologically distinct cell lines, we performed RNAi screens in each line with a set of double-stranded RNAs (dsRNAs) targeting 994 predicted cell shape regulators. Using automated fluorescence microscopy to visualize actin filaments, microtubules and DNA, we detected morphological phenotypes for 160 genes, one third of which have not been previously characterized in vivo. Genes with similar phenotypes corresponded to known components of pathways controlling cytoskeletal organization and cell shape, leading us to propose similar functions for previously uncharacterized genes. Furthermore, we were able to uncover genes acting within a specific pathway using a co-RNAi screen to identify dsRNA suppressors of a cell shape change induced by Pten dsRNA. CONCLUSIONS: Using RNAi, we identified genes that influence cytoskeletal organization and morphology in two distinct cell types. Some genes exhibited similar RNAi phenotypes in both cell types, while others appeared to have cell-type-specific functions, in part reflecting the different mechanisms used to generate a round or a flat cell morphology. PMID- 14527346 TI - Functional genomics of cell morphology using RNA interference: pick your style, broad or deep. AB - Several new studies have used RNA interference to screen for protein functions affecting cell shape, mitosis and cytokinesis of Drosophila cells in culture. One broad survey of nearly 1,000 proteins and three studies focused on cytoskeletal and motor proteins have identified key proteins essential for these processes in animal cells. PMID- 14527348 TI - An evaluation of completeness of tuberculosis notification in the United Kingdom. AB - BACKGROUND: There has been a resurgence of tuberculosis worldwide, mainly in developing countries but also affecting the United Kingdom (UK), and other Western countries. The control of tuberculosis is dependent on early identification of cases and timely notification to public health departments to ensure appropriate treatment of cases and screening of contacts. Tuberculosis is compulsorily notifiable in the UK, and the doctor making or suspecting the diagnosis is legally responsible for notification. There is evidence of under reporting of tuberculosis. This has implications for the control of tuberculosis as a disproportionate number of people who become infected are the most vulnerable in society, and are less likely to be identified and notified to the public health system. These include the poor, the homeless, refugees and ethnic minorities. METHOD: This study was a critical literature review on completeness of tuberculosis notification within the UK National Health Service (NHS) context. The review also identified data sources associated with reporting completeness and assessed whether studies corrected for undercount using capture-recapture (CR) methodology. Studies were included if they assessed completeness of tuberculosis notification quantitatively. The outcome measure used was notification completeness expressed between 0% and 100% of a defined denominator, or in numbers not notified where the denominator was unknown. RESULTS: Seven studies that met the inclusion and exclusion criteria were identified through electronic and manual search of published and unpublished literature. One study used CR methodology. Analysis of the seven studies showed that undernotification varied from 7% to 27% in studies that had a denominator; and 38%-49% extra cases were identified in studies which examined specific data sources like pathology reports or prescriptions for anti-tuberculosis drugs. Cases notified were more likely to have positive microbiology than cases not notified which were more likely to have positive histopathology or be surgical in-patients. Collation of prescription data of two or more anti-tuberculosis drugs increases case ascertainment of tuberculosis. CONCLUSION: The reporting of tuberculosis is incomplete in the UK, although notification is a statutory requirement. Undernotification leads to an underestimation of the disease burden and hinders implementation of appropriate prevention and control strategies. The notification system needs to be strengthened to include education and training of all sub specialities involved in diagnosis and treatment of tuberculosis. PMID- 14527347 TI - Independence of circadian entrainment state and responses to melatonin in male Siberian hamsters. AB - BACKGROUND: Seasonal fluctuations in physiology and behavior depend on the duration of nocturnal melatonin secretion programmed by the circadian system. A melatonin signal of a given duration, however, can elicit different responses depending on whether an animal was previously exposed to longer or shorter photoperiod signals (i.e., its photoperiodic history). This report examined in male Siberian hamsters which of two aspects of photoperiod history--prior melatonin exposure or entrainment state of the circadian system - is critical for generating contingent responses to a common photoperiodic signal. RESULTS: In Experiment #1, daily melatonin infusions of 5 or 10 h duration stimulated or inhibited gonadal growth, respectively, but had no effect on entrainment of the locomotor activity rhythm to long or short daylengths, thereby demonstrating that melatonin history and entrainment status could be experimentally dissociated. These manipulations were repeated in Experiment #2, and animals were subsequently exposed to a 12 week regimen of naturalistic melatonin signals shown in previous experiments to reveal photoperiodic history effects. Gonadal responses differed as a function of prior melatonin exposure but were unaffected by the circadian entrainment state. Experiment #3 demonstrated that a new photoperiodic history could be imparted during four weeks of exposure to long photoperiods. This effect, moreover, was blocked in animals treated concurrently with constant release melatonin capsules that obscured the endogenous melatonin signal: Following removal of the implants, the gonadal response depended not on the immediately antecedent circadian entrainment state, but on the more remote photoperiodic conditions prior to the melatonin implant. CONCLUSIONS: The interpretation of photoperiodic signals as a function of prior conditions depends specifically on the history of melatonin exposure. The photoperiodic regulation of circadian entrainment state contributes minimally to the interpretation of melatonin signals. PMID- 14527349 TI - Dynamic changes in blood cytokine levels as clinical indicators in severe acute respiratory syndrome. AB - OBJECTIVE: To investigate the dynamic changes observed in serum levels of interleukins (ILs), tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta1 (TGF-beta1) in severe acute respiratory syndrome (SARS) patients. METHODS: Sixty-one cases of SARS with positive antibodies to SARS coronavirus (SARS-CoV) were classified into the following categories: initial stage (3 - 7 days), peak stage (8 - 14 days), and remission and recovery stage (15 - 27 days). Forty-four healthy individuals were used as controls. Serum levels of ILs, TNF-alpha and TGF-beta 1 were measured in all subjects. Serum antibodies to SARS-CoV were detected only in SARS cases. RESULTS: The mean concentration of serum IL-6 in SARS patients did not differ from that in the control group in initial and peak stages, but became significantly higher in remission and recovery stage compared with the control group, initial and peak stages (P < 0.01). The mean concentration of serum IL-8 in SARS patients did not differ from that of the control group in initial stage, but was significantly higher than control group in peak stage and remission and recovery stage (P < 0.05). And it was more significantly higher in remission and recovery stage than in peak stage (P < 0.01). The mean concentrations of IL-16 and TNF-alpha in SARS patients were higher than those of the control group for every length of the clinical courses investigated, and were especially high in remission and recovery stage (P < 0.01). SARS patients experienced higher concentration of serum IL-13 compared with the controls in initial stage (P < 0.01), but returned to normal levels in peak stage and in remission and recovery stage. The mean concentration of serum IL-18 in SARS patients was significantly lower than that of the control group during all clinical courses (P < 0.05). The mean concentration of serum TGF beta1 in SARS patients was higher than that of the control group during all clinical courses. Although TGF-beta1 in serum decreased in remission and recovery stage in SARS patients, the average was still higher than that of the control group (P < 0.01). CONCLUSIONS: Most proinflammatory cytokines and TGF-beta 1 were elevated during the early phase of SARS, which may be associated with lung infiltration and proliferation. Concurrently, the mean concentration of serum IL 13 decreased gradually, and the mean concentration of serum IL-18 level in SARS patients was lower than that of the control group during all the courses of SARS, suggesting that the immune state of the patients with SARS was obviously abnormal. Observing the dynamic changes in blood cytokine levels can provide a scientific basis to assess pathogenesis and efficacy of clinical treatment of SARS. PMID- 14527350 TI - Severe acute respiratory syndrome-associated coronavirus genotype and its characterization. AB - OBJECTIVE: To study the severe acute respiratory syndrome (SARS)-associated coronavirus genotype and its characteristics. METHODS: A SARS-associated coronavirus isolate named ZJ01 was obtained from throat swab samples taken from a patient in Hangzhou, Zhejing province. The complete genome sequence of ZJ01 consisted of 29,715 bp (GenBank accession: AY297028, version: gi: 30910859). Seventeen SARS-associated coronavirus genome sequences in GenBank were compared to analyze the common sequence variations and the probability of co-occurrence of multiple polymorphisms or mutations. Phylogenetic analysis of those sequences was done. RESULTS: By bioinformatics processing and analysis, the 5 loci nucleotides at ZJ01 genome were found being T, T, G, T and T, respectively. Compared with other SARS-associated coronavirus genomes in the GenBank database, an A/G mutation was detected besides the other 4 mutation loci (C:G:C:C/T:T:T:T) involved in this genetic signature. Therefore a new definition was put forward according to the 5 mutation loci. SARS-associated coronavirus strains would be grouped into two genotypes (C:G:A:C:C/T:T:G:T:T), and abbreviated as SARS coronavirus C genotype and T genotype. On the basis of this new definition, the ZJ01 isolate belongs to SARS-associated coronavirus T genotype, first discovered and reported in mainland China. Phylogenetic analysis of the spike protein gene fragments of these SARS-associated coronavirus strains showed that the GZ01 isolate was phylogenetically distinct from other isolates, and compared with groups F1 and F2 of the T genotype, the isolates of BJ01 and CUHK-W1 were more closely related to the GZ01 isolate. It was interesting to find that two (A/G and C/T) of the five mutation loci occurred in the spike protein gene, which caused changes of Asp to Gly and Thr to Ile in the protein, respectively. CONCLUSION: Attention should be paid to whether these genotype and mutation patterns are related to the virus's biological activities,epidemic characteristics and host clinical symptoms. PMID- 14527351 TI - Ventilation of wards and nosocomial outbreak of severe acute respiratory syndrome among healthcare workers. AB - OBJECTIVE: To identify valid measures for preventing outbreaks of severe acute respiratory syndrome (SARS) among protected healthcare workers in isolation units. METHODS: Architectural factors, admitted SARS cases and infection of healthcare workers in different isolation wards between January 30 and March 30, 2003 were analyzed. RESULTS: Four types of isolation wards were analyzed, including the ward where the thirty-first bed was located on the twelfth floor, the laminar flow ward in the Intensive Care Unit where the tenth bed was located on the fifteenth floor, the ward where the twenty-seventh bed was located on the thirteenth floor of the Lingnan Building, and thirty wards on the fourteenth to eighteenth floors of the Zhongshan Building. The ratios (m(2)/m(3)) of the area of the ventilation windows to the volume of the rooms were 0, 0, 1:95 and 1:40, respectively. Numbers of SARS cases in the wards mentioned above were 1, 1, 1 and 96, respectively. Total times of hospitalization were 43, 168, 110 and 1272 hours, respectively. The infection rates of the healthcare workers in the areas mentioned above were 73.2%, 32.1%, 27.5% and 1.7%, respectively. The difference in the infection rates was of statistical significance. CONCLUSIONS: Isolating SARS cases in wards with good ventilation could reduce the viral load of the ward and might be the key to preventing outbreaks of SARS among healthcare workers along with strict personal protection measures in isolation units. PMID- 14527352 TI - Chromosomal changes detected by fluorescence in situ hybridization in patients with acute lymphoblastic leukemia. AB - OBJECTIVES: To investigate patients with acute lymphoblastic leukemia (ALL) for TEL/AML1 fusion, BCR/ABL fusion, MLL gene rearrangements, and numerical changes of chromosomes 4, 10, 17 and 21 by fluorescence in situ hybridization (FISH) and to determine the relationship and the significance of those findings. METHODS: Fifty-one American patients (34 men and 17 women) were included in this study. Of them there were 41 patients with pro-B cell type ALL, 9 with B cell type ALL and 1 with T cell type ALL. Chromosome metaphases of each sample were prepared according to standard protocols. Fluorescence in situ hybridization was performed using commercially available DNA probes, including whole chromosome painting probes, locus specific probes, specific chromosome centromere probes and dual color/multiple color translocation fusion probes. The digital image analysis was carried out using Cytovision and Quips FISH programs. RESULTS: An overall incidence of chromosomal anomalies, including t (9;22), MLL gene rearrangements, t (12;21), and numerical chromosomal anomalies of chromosomes 4, 10, 17 and 21 was found in 33 patients (65%). Thirty-one of them were pediatric patients and two adults. The t (12;21) was the commonest chromosomal anomaly detected in this population; 14 out of the 45 pediatric patients (31%) were positive for TEL/AML1 fusion, among which three had an additional derivative 21 [t (12;21)], four had a deletion of 12p and two had an extra copy of chromosome 21. All 14 patients with positive TEL/AML1 fusion had ALL pre-B cell or B-cell lineage according to standard immunotyping. The percentage of cells with fusion signals ranged from 20% to 80%. All fourteen patients positive for TEL/AML1 gene fusion were mosaic. Three out of the 14 patients positive for the TEL/AML1 gene fusion were originally reported to be culture failures and none of the remaining eleven samples had been found to have chromosome 12 abnormalities by conventional cytogenetic techniques. All pediatric patients with pre-T or T cell lineage and the six adults were negative for TEL/AML1 fusion. One patient had double Philadelphia chromosomes, three had a rearrangement or a deletion of the MLL gene, one had t (4;11) and two had a deletion of the MLL. One of the patients with an MLL deletion also had a large ring of chromosome 21, and r (21) was caused by AML1 gene tandemly duplicated at least five times. The second case with the MLL deletion was also unique, the patient had a t (12;21) as well. A total of 20 patients had numerical changes (gain or loss) of chromosomes 4, 10, 17 and 21. Eight patients were found to have trisomies of three or four different chromosomes. Interestingly, seven of these patients did not have TEL/AML1, BCR/ABL or the MLL gene rearrangement; one did have the TEL/AML1 gene fusion. Eleven patients with pro-B cell or B cell type ALL (9 children with ALL, 2 adults with ALL) had numerical changes of chromosome 21 (gain 1 or 2 chromosome 21), among them, 10 patients had no structural alteration of chromosome 21, and one was combined by t (12; 21). Four patients had a monosomy of chromosome 17 and three out of these patients with monosomy 17 also had a fusion signal of TEL/AML1. CONCLUSIONS: FISH plays an important role in detecting chromosome changes, especially in some cryptic chromosome translocations and patients with culture failures. This study found a trend towards a division between patients who had structural changes such as t (12;21) or a ring chromosome 21 and those who had numerical changes of chromosome 21 as well as the patients with TEL/AML1 fusion and patients with the coexistence of numerical chromosomal changes of chromosomes 4, 10 and 17. In our opinion there are two separate mechanisms which lead to the development or progression of leukemia. PMID- 14527353 TI - Prevalence of antimicrobial resistance of Streptococcus pneumoniae in Chinese children: four hospitals surveillance. AB - OBJECTIVE: To investigate the nasal carriage of antibiotic-resistant pneumococci in children of < 5 years old in the following four cities, Beijing, Shanghai, Guangzhou and Xi'an. METHODS: A total of 647 pneumococci strains were isolated and detected. Minimal inhibition concentrations (MICs) of antibiotics were determined by E-test. Disk diffusion test was used for the measurement of antimicrobial susceptibility. RESULTS: Prevalence of penicillin non-susceptible Streptococcus pneumoniae in the four cities was 41%, with Guangzhou (60.8%) ranking first, followed by Xi'an (45%), Shanghai (37%) and Beijing (25.9%). The majority of penicillin non-susceptibility isolates (23.9% - 53.8%) had a low level of resistance (MIC 0.64 - 1.5 microg/ml). The most sensitive antimicrobials in terms of percentage of susceptible organisms were amoxicillin-clavulanic acid (99.4%), followed by ceftriaxone (92.1%); cefurxime and cefaclor were slightly more sensitive than penicillin with susceptibility of 74.8% and 77.9%. Erythromycin, tetracycline and TMP-SMZ were highly resistant (83.6%, 82.1% and 76.2% respectively). Among erythromycin resistant isolates, 100% were resistant to azithromycin, 98.6% to clarithromycin, 97.2% to roxithromycin and spiramycin, and 96.6% to clindamycin. 97.2% (141/145) were typical of the macrolides lincosamides-streptogramons B (MLSB) resistance phenotype, and 2.8% (4/145) were M phenotype. The group of PRSP was with significantly higher rates of non susceptibility for ceftriaxone (18.4%), cefurxime (58.6%), cefaclor (53.4%), compared with the group of PEN-S (0.5%, 1.8% and 0.2%, respectively) and the rate of multi-drug resistance in the isolates of PRSP group (92.9%) was significantly higher than that of PEN-S group (59.2%). CONCLUSION: The rates of penicillin and multi-drug resistance among isolates of pneumococci carried nasally in are high children and the high prevalence of multi-drug resistance in the Chinese population may be becoming one of the most serious problems in this century. PMID- 14527354 TI - A possible receptor for beta 2 glycoprotein I on the membrane of hepatoma cell line smmc7721. AB - OBJECTIVES: To study the interaction of beta-2-glycoprotein I (beta 2GPI) with the membrane of hepatocytes and determine whether beta 2GPI participates in HBV infection. METHODS: Ligand blotting, fluorescence microscopy, and fluorescence activated cell sorter (FACS) analysis were used to detect the specific interaction of beta 2GPI with the hepatoma cell line smmc7721, the gastric carcinoma cell line SGC7901, and the lymphoma cell line HL-60. RESULTS: A specific 40 kDa beta 2GPI band was observed by ligand blotting in the case of smmc7721 cells. No such band was observed in SGC7901 or HL-60 cells. Fluorescence microscopy also revealed specific binding of FITC-beta 2GPI to smmc7721 cells, but neither to SGC7901 nor HL-60 cells. FACS analysis demonstrated that the binding rate of FITC-beta 2GPI to smmc7721 cells was significantly higher than these in SGC7901 and HL-60 cells (P < 0.01). The binding rate to smmc7721 cells did not increase with increasing amounts of FITC-beta 2GPI. CONCLUSIONS: There is a specific beta 2GPI-binding protein on the membrane of hepatoma cells in cell line smmc7721 which as the beta 2GPI receptor may participate in HBV infection of hepatocytes. PMID- 14527355 TI - Clinical and histological characteristics of chronic hepatitis B with negative hepatitis B e-antigen. AB - OBJECTIVE: To study the clinical and histological features of chronic hepatitis B (CHB) with negative hepatitis B e-antigen (HBeAg). METHODS: A total of 743 in patients with chronic hepatitis B were recruited into the study and divided into two groups according to the HBeAg status. The correlation among alanine transaminase (ALT) levels, hepatitis B virus (HBV) DNA semiquantification, and the liver histopathological data were detected. RESULTS: Of the 743 successive in patients, 267 (35.9%) were HBeAg-negative. The HBDAG-negative group had significantly lower serologic HBV DNA levels (63.0% of < 100 pg/ml) vs HBeAg positive (42.6%, P < 0.001), while more sever inflammation (58.1% of inflammatory scores of histological activity index (HAIinf > or = 9) vs HBeAg-positive group (46.0%, P < 0.001) and severe fibrosis (45.3% of fibrosis scores of histological activity index (HAIfib > or = 3) vs HBeAg-positive group (27.9%, P < 0.001) of liver histology. In HBeAg-positive patients, increasing ALI levels were significantly associated with high inflammation and fibrosis scores and low HBV DNA levels. However, it was not the case in the HBeAg-negative cases. In HBeAg positive patients, 91.3% of them had HAIinf > or = 9 and 65.7% had HAIfib > or = 3 with HBV DNA > 100 pg/ml, while 8.2% of them had HAIinf > or = 9 and 12.3% had HAIfib > or = 3 with HBV DNA < 20 pg/ml, indicating an obverse correlation between HBV DNA levels and histology scores. CONCLUSIONS: As regards clinical and histological background, the chronic HBeAg-negative hepatitis B is a different subpopulation from the HBeAg-positive counterpart. PMID- 14527356 TI - Preliminary research on the co-infection of human immunodeficiency virus and hepatitis virus in intravenous drug users. AB - OBJECTIVE: To confirm the close relationship of high co-infection rate between HIV and hepatitis virus in intravenous drug users (i.v.DUs). METHODS: Anti-HIV, HBV and HCV were detected by ELISA in the serum from 35 scattered and 15 massed i.v.DUs. PCR and RT-PCR were performed to confirm the infection of HIV, HBV, HCV, HGV and TTV among the 15 massed intravenous drug abusers. RESULTS: Among the 50 i.v.DUs, the positive rates of anti-HCV1 HBsAg, anti-HBe and anti-HBc were 92% (46/50), 12% (6/50), 10% (5/50) and 66% (33/50), respectively. In the samples of HBsAg positive, their HBeAg was also positive. Although the positive rate of serum markers was different in the massed i.v.DUs compared to the scattered i.v.DUs, no significant difference was shown. In the cases of massed i.v.DUs, the positive rates of HIV DNA, HBV-DNA, HCV-RNA, HGV-RNA, and TTV-DNA were 100% (15/15), 26.6% (4/15), 53.3% (8/15), 33.3% (5/15) and 26.6% (4/15), respectively. Among the 15 massed intravenous drug users, one was infected with HIV, HBV, HCV, HGV and TTV; two were infected with HIV, HBV, HCV and HGV; three were infected only with HIV; and the remaining had other forms of co-infection. CONCLUSION: The co-infection rate of HIV, HBV, HCV, HGV and TTV in intravenous drug users is very high. PMID- 14527357 TI - Apoptosis in Raji cell line induced by influenza A virus. AB - OBJECTIVE: To study the apoptotic effects of influenza A virus on the Raji cell line. METHODS: Cultured Raji cells were infected with influenza A virus at a multiplicity of infection (m.o.i) of 20 and the effects of apoptosis were detected at different time points post infection using the following methods: electron microscope, DNA agarose gel electrophoresis, PI stained flow cytometry (FCM) and Annexin-V FITC/PI stained FCM. RESULTS: Raji cells infected with influenza A virus showed changes of morphology apoptosis, DNA agarose electrophoresis also demonstrated a ladder-like pattern of DNA fragments in a time-dependent manner. PI stained FCM showed "apoptosis peak" and FITC/PI stained FCM showed apoptotic cells. Quantitative analysis indicated that the percentage of apoptotic Raji cells increased after infection, and cycloheximide (CHX), an eukaryotic transcription inhibitor, could effectively inhibit the apoptotic effects of influenza A virus in vitro. CONCLUSIONS: Influenza A virus can induce apoptosis in Raji cell line suggesting that it may lead to a potential method for tumor therapy. PMID- 14527358 TI - Transesophageal cardioversion of atrial flutter and atrial fibrillation using an electric balloon electrode system. AB - OBJECTIVE: To determine the feasibility and efficiency of terminating atrial flutter (AFL) and atrial fibrillation (AF) using synchronous low-energy shocks delivered through a novel transesophageal electric balloon electrode system. METHODS: By using a novel electric balloon electrode system, we attempted 91 transesophageal cardioversions in 52 patients, to treat 53 episodes of AFL and 38 episodes of AF. RESULTS: Of the 40 patients of AFL that failed to respond to drug therapy, 37 (92.5%) were successfully countershocked to sinus rhythm by transesophageal cardioversion, with a mean energy of (22.70 +/- 4.50) J (20 - 30 J). Of the 19 patients in AF, transesophageal cardioversion was successful in 16 (84.2%) cases, requiring a mean delivered energy of (17.38 +/- 8.58) J (3 - 30 J). There were no complications such as heart block or ventricular fibrillation, and no evidence of esophageal injury. CONCLUSIONS: Transesophageal cardioversion using an electric balloon electrode system is an effective and feasible method for the treatment of AFL and AF. It requires low energy and no anesthesia, leads to less trauma, and shows a high cardioversion success rate that may prove valuable in the management of tachyarrhythmias. PMID- 14527359 TI - Identification of genes expressed during myocardial development. AB - OBJECTIVE: To identify genes expressed in the fetal heart that are potentially important for myocardial development and cardiomyocyte proliferation. METHODS: mRNAs from fetal (29 weeks) and adult cardiomyocytes were use for suppression subtractive hybridization (SSH). Both forward (fetal as tester) and reverse (adult as driver) subtractions were performed. Clones confirmed by dot-blot analysis to be differentially expressed were sequenced and analyzed. RESULTS: Differential expressions were detected for 39 out of 96 (41%) clones on forward subtraction and 24 out of 80 (30%) clones on reverse. For fetal dominating genes, 28 clones matched to 10 known genes (COL1A2, COL3A1, endomucin, HBG1, HBG2, PCBP2, LOC51144, TGFBI, vinculin and PND), 9 clones to 5 cDNAs of unknown functions (accession AK021715, AF085867, AB040948, AB051460 and AB051512) and 2 clones had homology to hEST sequences. For the reverse subtraction, all clones showed homology to mitochondrial transcripts. CONCLUSIONS: We successfully applied SSH to detect those genes differentially expressed in fetal cardiac myocytes, some of which have not been shown relative to myocardial development. PMID- 14527360 TI - Mutation analysis of potassium channel genes KCNQ1 and KCNH2 in patients with long QT syndrome. AB - OBJECTIVE: To determine mutations of two common potassium channel subunit genes KCNQ1, KCNH2 causing long QT syndrome (LQTS) in the Chinese. METHODS: Thirty-one Chinese LQTS pedigrees were characterized for mutations in the two LQTS genes, KCNQ1 and KCNH2, by sequencing. RESULTS: Two novel KCNQ1 mutations, S277L in the S5 domain and G306V in the channel pore, and two novel KCNH2 mutations, L413P in the transmembrane domain S1 and L559H in the transmembrane domain S5 were identified. The triggering factors for cardiac events developed in these mutation carriers included physical exercise and excitation. Mutation L413P in KCNH2 was associated with the notched T wave on ECGs. Mutation L559H in KCNH2 was associated with the typical bifid T wave on ECGs. Mutation S277L in KCNQ1 was associated with a high-amplitude T wave and G306V was associated with a low amplitude T wave. Two likely polymorphisms, IVS11 + 18C > T in KCNQ1 and L520V in KCNH2 were also identified in two LQTS patients. CONCLUSIONS: The mutation rates for both KCNQ1 (6.4%) and KCNH2 (6.4%) are lower in the Chinese population than those from North America or Europe. PMID- 14527361 TI - Expression of platelet-endothelial cell adhesion molecule-1 in human umbilical vein endothelial cells by exposure to advanced glycosylation end products and inflammatory mediators. AB - OBJECTIVE: To determine whether advanced glycosylation end products modified bovine serum albumin (AGEs-BSA) affects endothelial cell lateral junction protein, platelet-endothelial cell adhesion molecule-1 (PECAM-1) in the presence or absence of inflammatory mediators. METHODS: Cultured human umbilical vein endothelial cells (HUVECs) were exposed to AGEs-BSA for 6, 12, 24, and 36 hours, and exposed to AGEs-BSA glycosylated with different concentrations of glucose, tumor necrosis factor-alpha (TNF-alpha), interferon (IFN-gamma), TNF-alpha + IFN gamma and AGEs-BSA + TNF-alpha for 24 hours, respectively. Expression of PECAM-1 mRNA was measured by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) with beta-actin as an internal standard, and sequencing of RT PCR products was performed to confirm the specificity of amplification for PECAM 1 gene. The endothelial cell surface expression of PECAM-1 was determined by flow cytometry (FCM). RESULTS: There were no significant changes in the expression of PECAM-1 mRNA and protein when the cells were exposed to AGEs-BSA with different concentrations or periods (P > 0.05). However, PECAM-1 expression was reduced in the cells treated with TNF-alpha, IFN-gamma, TNF-alpha + IFN-gamma and AGEs-BSA + TNF-alpha. The level of PECAM-1 treated with AGEs-BSA + TNF-alpha was lower than that of TNF-alpha treated alone (P < 0.01). CONCLUSIONS: AGEs-BSA had no effect on the expression of PECAM-1 mRNA and protein in cultured HUVEC. With the presence of inflammatory mediator TNF-alpha, AGEs-BSA decreased the level of PECAM-1, which might reduce the adhesion interaction between adjacent endothelial cells, enhance the permeability of endothelial cells, and might be implicated in the endothelial dysfunction and pathogenesis of atherosclerosis in patients with diabetes mellitus. The significance of this phenomenon in intracellular signal transduction remains to be determined. PMID- 14527362 TI - Effects of different therapies on coronary artery disease. AB - OBJECTIVE: To analyse the effects of different therapies on coronary artery disease (CAD). METHODS: A total of 1055 patients who suffered from CAD diagnosed by coronary angiography were divided into three groups, namely pure drug therapy, percutaneous coronary intervention (PCI) and coronary artery bypass graft (CABG) groups. Follow up was carried out from March to May in 2001, and the major adverse cardiac events (MACEs) including death, no-lethal myocardial infarction (MI) and revascularization were observed. In long-term observation, angina reoccurred, and their improvement was evaluated. The short-term period was defined as the duration of 30 days after discharge, and the long term period was defined as the duration from 30 days after discharge. RESULTS: In the long-term period, the recurrences of angina both in PCI group and CABG group were lower than pure drug group (P 0.018, 0.002 respectively). No differences about long term endpoint events were observed among these three groups (P > 0.05). Forty-two patients suffering from left main coronary disease were intervened by the three therapies, and there was no death or MI both in PCI and CABG groups, three patients died and suffered from AMI in pure drug therapy group (P = 0.015). In the short-term period, mortality in CABG group (5.77%) was higher than those in the other two groups (1.91% for PCI, and 1.40% for medical therapy, P = 0.002), and no obvious difference observed in the latter two groups. No significance was concluded about the recent MI among this three groups (P = 0.357). There were no differences on revascularization in these three groups. CONCLUSIONS: Percutaneous coronary interventions can not only reduce the attack of angina but also improve the life quality of patients, however it can not improve the long-term existence but left main CAD. PMID- 14527363 TI - Inhibitory effect of tea polyphenols on renal cell apoptosis in rat test subjects suffering from cyclosporine-induced chronic nephrotoxicity. AB - OBJECTIVE: To investigate the inhibitory effect of tea polyphenols on renal cell apoptosis in rat test subjects suffering from cyclosporine A (CsA)-induced chronic nephrotoxicity. METHODS: Four groups of rats with CsA-induced chronic nephrotoxicity were respectively treated with vehicle olive oil, tea polyphenols, CsA and tea polyphenols plus CsA. At the end of the 28th day of treatment, 24 hours urine and blood samples were obtained, and the animals were then sacrificed. The serum and urine samples were analysed for creatinine clearance, and kidney tissue was used for pathologic analysis of renal tubular injury and interstitial fibrosis. The TUNEL assay, apoptosis-related enzyme caspase-3 mRNA detected by RT-PCR, and its enzymatic activity were analysed for the possible detections of cell apoptosis. RESULTS: CsA-treated rats displayed increased apoptosis of the tubular and interstitial cells, in comparison with vehicle treated controls (18.3 +/- 4.6 vs 4.8 +/- 1.3 cells/mm(2), P < 0.05). In comparison with animals treated by CsA, animals treated with CsA plus tea polyphenols demonstrated significantly improved levels of creatinine clearance (0.12 +/- 0.03 vs 0.22 +/- 0.02 ml.min(-1).100 g(-1) body weight, P < 0.05), tubular injury (2.29 +/- 0.43 vs 1.42 +/- 0.26, P < 0.05), and interstitial fibrosis (2.83 +/- 0.20 vs 1.46 +/- 0.19, P < 0.05), and showed a statistically significant decrease in tubular and interstitial cell apoptosis (18.3 +/- 4.6 vs 7.7 +/- 2.1 cells/mm(2), P < 0.05). The expression of caspase-3 mRNA and caspase 3 activity was significantly higher in the CsA-treated group than that of the CsA plus tea polyphenols (TP)-treated group (P < 0.05). CONCLUSION: These results suggested that tea polyphenols significantly inhibits apoptosis of the tubular and interstitial cells in rats with cyclosporine-induced chronic nephrotoxicity, and that tea polyphenols may be useful to prevent CsA-associated kidney toxicity. PMID- 14527364 TI - Immortalization of human articular chondrocytes and induction of their phenotype. AB - OBJECTIVE: To immortalize human articular chondrocytes (HACs) using gene transfection and to maintain stable phenotype of transformed HACs after induction. METHODS: HACs were transfected with the retroviral vector pLXSN encoding human papillomavirus 16E7 (HPV16E7), and the transformed clones were sorted and proliferated. Karyotype analysis, clone forming tests and nude mice tumor forming tests were applied to check the characteristics of the transformation. Type II collagen of transformed chondrocytes was inducted with free serum medium (FSM) supplemented with nutridoma-sp and ascorbate. RESULTS: Immortalized HACs were isolated with fifty passages achieved. The HPV16E7 transformed cells were confirmed to be benign. Induction of FSM with nutridoma-sp and ascorbate promoted type II collagen of transformed chondrocytes to the high levels of normal chondrocytes. CONCLUSION: HACs transformed with HPV16E7 survive for long periods in vitro, and type II collagen can maintain stability after induction. PMID- 14527365 TI - Effects of dexamethasone on proliferation, differentiation and apoptosis of adult human osteoblasts in vitro. AB - OBJECTIVE: To observe the effects of dexamethasone on proliferation, differentiation and apoptosis of adult human osteoblasts in vitro. METHODS: Iliac trabecular bone specimens were obtained from adult patients undergoing necessary surgery. After the bone pieces were digested with collagenase-trypsin, osteoblasts were released and incubated at 37 degrees C in a relative humidity of 95% and 5% CO2. Then, the cells were purified, and their passages were given DMEM F12 and fetal bovine serum medium. Subsequently, 10(-8) mol/L dexamethasone was added into the culture medium to incubate the osteoblasts for three days, and the cells from control groups were incubated without any drugs. All cells were observed continually with phase contrast microscope and transmission electron microscope. Finally, apoptosis was detected by the use of terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) and biochemical indices, alkaline phosphatase (ALP) and osteocalcin (OCN) were used to determine the effects of dexamethasone on proliferation, differentiation and apoptosis of adult osteoblasts in vitro. RESULTS: In the adult osteoblasts obtained by collagenase-trypsin digestion, it achieved high survival, stable biochemical indices and excellent purification. Under the condition of dexamethasone 10(-8) mol/L and osteoblasts 10,000/ml, there was significant promotion of ALP and OCN secretion without cell apoptosis. CONCLUSIONS: Dexamethasone has a significant effect on the proliferation and differentiation of adult osteoblasts in vitro without apoptosis, and dexamethasone at the suggested concentration can be used as positive control in drug studies for osteoporosis treatment. PMID- 14527366 TI - Effects of subconvulsive electrical stimulation to the hippocampus on emotionality and spatial learning and memory in rats. AB - OBJECTIVE: To observe the effects of repeated subconvulsive electrical stimuli to the hippocampus on the emotional behavior and spatial learning and memory ability in rats. METHODS: One hundred and eight male Wistar rats were randomized into 3 groups. Animals in group SE (n = 42) were given subconvulsive electrical stimulation to the hippocampus through a constant pulsating current of 100 mu A with an intratrain frequency of 25 Hz, pulse duration of 1 millisecond, train duration of 10 seconds and interstimulus interval of 7 minutes, 8 times a day, for 5 days. In the electrode control group or CE group (n = 33), animals were implanted with an electrode in the hippocampus, but were not stimulated. Group NC (n = 33) animals received no electrode or any stimulation. The emotional behavior of experimental rats was examined by activity in an unfamiliar open field and resistance to capture from the open field, while the spatial learning and memory ability was measured during training in a Morris water maze. RESULTS: The stimulated rats tested 1 month after the last round of stimulation displayed substantial decreases in open field activity (scale: 10.4 +/- 2.3, P < 0.05) and increases in resistance to capture (scale: 2.85 +/- 0.56, P < 0.01). The amount of time for rats in group SE to find the platform (latency) as a measurement for spatial bias was prolonged (29 +/- 7) seconds after 15 trials in the water maze, P < 0.05). The experimental rats swam aimlessly in all four pool quadrants during the probe trial in the Morris water maze. CONCLUSIONS: Following repeated subconvulsive electrical stimuli to the hippocampus, rats displayed long-lasting significant abnormalities in emotional behavior, increased anxiety and defensiveness, enhanced ease to and delayed habituation to startlement, transitory spatial learning and memory disorder, which parallels many of the symptoms in posttraumatic stress disorder patients. PMID- 14527367 TI - Inhibition of lovastatin on proliferation and expression of proinflammatory cytokines in cultured human glomerular mesangial cells. AB - OBJECTIVE: To study the effects and mechanism of lovastatin on cell proliferation and expression of proinflammatory cytokines in cultured human glomerular mesangial cells. METHODS: The influence of lovastatin on HMC proliferation was evaluated with 3H-thymidine incorporation. mRNA expression of proinflammatory cytokines (IL-1 beta, IL-6, TNF-alpha, and MCP-1) and activation of NF-kappa B of HMC were measured using Reverse transcription-polymerase chain reaction (RT-PCR) and electrophoretic mobility shift assay (EMSA) respectively. RESULTS: Lovastatin was found to have inhibitory effects on human mesangial cell (HMC) proliferation and lipopolysaccharide (LPS)-mediated human mesangine cell HMC mRNA expression of proinflammatory cytokines via activation of NF-kappa B. The effect of lovstatin on HMC could be prevented when the mevalonate and farnesol were added to the culture. CONCLUSION: Lovastatin may decrease HMC proliferation and production of proinflammatory cytokines through the inhibition of NF-kappa B activation. This provided experimental evidence for further evaluation of the renal protective effect of HRI, suggesting that it may be a potent agent for prevention of progressive renal diseases aside from its lipid-lowering effect. PMID- 14527368 TI - Expression of aquaporin-1 in the human peritoneum and the effect of peritoneal dialysis on its expression. AB - OBJECTIVE: To investigate the expression of aquaporin-1 (AQP1) in the human peritoneum and to evaluate the effect of peritoneal dialysis (PD) on its expression. METHODS: Peritoneal biopsies were obtained from normal subjects (n = 10), uremic nondialysis patients (n = 12) at catheter insertion and PD patients (n = 10) at the time of catheter removal, reinsertion or renal transplantation. Western blot, immuno-histochemical staining and reverse transcript-polymerase chain reaction (RT-PCR) techniques were used to investigate AQP1 expression. RESULTS: All peritoneal samples expressed AQP1 at both mRNA and protein levels. Western blot revealed a major band at 28 kD as well as more diffuse bands between 35 and 50 kD. The 28 kD band represents the nonglycosylated form of the protein while the 35 - 50 kD bands correspond to glycosylated AQP1. Immunohistochemical staining found the positive deposits were distributed in the mesothelial cells, endothelial cells of capillaries, venules and small veins, whereas no signal was detected in the arterioles. Semi-quantitative analysis showed that AQP1 expression was remarkably stable in all samples, whatever their origin (P > 0.05). CONCLUSIONS: Our findings suggested that AQP1 is the molecular counterpart of an ultra small pore during PD. Secondly, the peritoneal mesothelial cell might also be involved in peritoneal transcellular water transport. As regards whether or not the structural or distributional alterations of AQP1 in the peritoneum may be more obviously expressed during PD, further study is needed. PMID- 14527369 TI - The relationship between heart rate variability and prostaglandin E2 in patients with renal insufficiencies. AB - OBJECTIVE: To investigate the relationship between heart rate variability (HRV) and prostaglandin E2 (PGE2) in patients with renal insufficiencies. METHODS: HRV blood and 24-hour urine prostaglandin E2 (PGE2) detection were detected in the following 4 groups of people: group A was a control group comprised of 20 normal individuals; group B had 20 patients with renal disease but exhibiting normal renal function; group C contained 20 patients with renal disease and compensatory renal function; group D had 20 patients demonstrating renal insufficiencies. The indices standard deviation of all NN intervals (SDNN), index of standard deviation of the averages of NN intervals (SDANN), mean of the standard deviation of all NN intervals performed on all 5-minute segments of the entire recording (SDNNindex), square root of the mean of the sum of the squares of differences between adjacent NN intervals (rMSSD) and NN50 count divided by the total number of all NN intervals (PNN50) were used to evaluate HRV, blood and 24-hour urine were determined by specific radioimmunoassay (RIA). RESULTS: PGE2 in blood and urine and HRV exactly within 24 hours in patients with renal insufficiencies, negatively correlated with worsening damage to renal function. There was a slight or moderate correlation between blood and urine PGE2 and the SDNNindex, SDANNindex, SDNN, rMSSD and PNN50 indices (P < 0.05). CONCLUSIONS: HRV and cardiac autonomic regulatory functions are decreased in the patients with renal insufficiencies, while lower levels of PGE2 may be a related factor. PMID- 14527370 TI - Pathological demography of native patients in a nephrology center in China. AB - OBJECTIVE: To analysis the pathological demography in Chinese patients undergoing renal biopsy from our nephrology center. METHODS: Between January 1979 and October 2000 in Jinling Hospital, Nanjing, China, 10,002 attempts of percutaneous renal were performed in patients with renal disease from 33 provinces of China. The pathological classifications were made according to the WHO criteria of 1982 for renal pathology or the modified WHO criteria of 1995 by a panel of pathologists and nephrologists during routine clinical-pathological rounds. The pathological demography between those specimens collected from 1979 - 1989 and those from 1990 - 1999 was compared. RESULTS: The mean age of the 10,002 subjects undergoing renal biopsy was 31.4 +/- 13.0 years (ranging from 1 to 78 years), with a male to female ratio of 1.3:1; for the 592 renal transplant recipients, the mean age was 37.5 +/- 9.1 years (ranging from 16 to 66 years), with a male to female ratio of 2.36:1. Primary glomerular diseases (PGD) accounted for 71% of the total patients undergoing renal biopsies, secondary glomerular nephritis (SGN) 23%, tubular-interstitial diseases 3.2%, unclassified renal diseases 1.3%, hereditary and congenital renal diseases 1.0%, end stage renal diseases 0.96%, and recently realized or rare renal diseases 0.15%. IgA nephropathy (IgAN) was the most frequent pathological pattern (40%) of PGD, followed by mesangial proliferative lesion (MsPL) (30%), membranous nephropathy (MN) (10%), and focal segmental glomerulosclerosis (FSGS) (6%). Lupus nephritis (LN) was the most pathology common seen (74%) in SGN. During the 22 years of the study period, there was a steady increase in patients with SGN discovered during pathological evaluation of renal disorders. A rise in prevalence was found in IgA nephropathy, MN (both P < 0.001), crescentic glomerulonephritis (P < 0.0001), anti-GBM disease, and hemolytic-uremic syndrome/thrombotic thrombocytopenic purpura related renal damages (both P < 0.001). There was a decrease in endocapillary proliferative glomerulonephritis (P < 0.001) and IgM nephropathy (IgMN) (P < 0.01) from 1990 - 1999 as compared to 1979 - 1989. Infrequent renal pathological entities were also diagnosed in this group, including Niemann Pick disease, Fabry's disease, POEMS syndrome, and lipoprotein glomerulonephropathy. CONCLUSIONS: This is the largest series of renal biopsy data in China, and therefore may reflect the demographic picture of renal diseases in this country. Changes in prevalence of renal pathological entities were reflected in this group of patients over the last 22 years. In primary glomerular diseases, IgA nephropathy is still the most frequently observed pathological pattern. In SGN, LN appeared the most often. Increased prevalence was found in anti-GBM nephritis and HUS/TTP. PMID- 14527371 TI - Shenfu injection reduces toxicity of bupivacaine in rats. AB - OBJECTIVE: To investigate the effects of injecting Shenfu, an extract of traditional Chinese herbal medicines, on the central nervous system (CNS) and the cardiac toxicity of bupivacaine in rats. METHODS: Sixteen male Sprague-Dawley rats, weighing form 280 to 320 g, were randomly assigned to two groups (n = 8 in each group). Animals in the control group received a saline injection 10 ml/kg while animals in the Shenfu group received an injection of Shenfu 10 ml/kg intraperitoneally 30 minutes before intravenous infusion of bupivacaine. Lead II of an electrocardiogram (EEG) was continuously monitored after 3 needles were inserted into the skin of both forelimbs and the left hind-leg of each rat. The femoral artery was cannulated for measurement of arterial blood pressure and blood sampling. The femoral vein was cannulated for the infusion of bupivacaine. After baseline measurement (arterial blood pressure, heart rate and arterial blood gas), 0.5% bupivacaine was infused intravenously at a rate of 2 mg.kg( 1).min(-1) to all animals until asystole occurred. The time of bupivacaine induced convulsions, arrhythmia and asystole were determined. The dose of bupivacaine was then calculated at the corresponding time point. RESULTS: The doses of bupivacaine that induced convulsions, arrhythmia and cardiac arrest were remarkably larger in Shenfu injection-treated animals than in saline-treated rats [convulsions, (10.5 +/- 1.9) mg/kg vs (7.2 +/- 1.5) mg/kg; arrhythmia (10.5 +/- 2.0) mg/kg vs (7.2 +/- 1.9) mg/kg; asystole, (32.8 +/- 8.5) mg/kg vs (25.0 +/- 5.0) mg/kg; P = 0.006, 0.009 and 0.044, respectively]. CONCLUSION: The Shenfu injection is able to reduce the toxicity of bupivacaine to CNS and cardiac system in rats. PMID- 14527372 TI - Comparison of remifentanil and fentanyl in patients undergoing modified radical mastectomy or total hysterectomy. AB - OBJECTIVE: To compare the efficacy and safety of remifentanil and fentanyl in patients undergoing a modified radical mastectomy or total hysterectomy. METHODS: Fifty-four patients were evenly randomised into remifentanil group and fentanyl group. Anesthesia was induced by propofol (1 - 2 mg/kg) and either remifentanil (2 microg/kg) or fentanyl (2.5 microg/kg), and was maintained with inhalation of nitrous oxide in oxygen (2:1) and continuous infusion of either remifentanil (0.2 microg.kg(-1).min(-1)) or fentanyl (0.03 microg.kg(-1).min(-1)). RESULTS: The number of patients exhibiting light anesthesia responses in the remifentanil group during intubation and the maintenance of anesthesia was significantly less than that in the fentanyl group. Both systolic and diastolic blood pressures in the fentanyl group were significantly higher than those in the remifentanil group during intubation, skin incision, maintenance of anesthesia and extubation. The time to opening eyes on command and the time for extubation after surgery was comparable between the two groups. More patients in the remifentanil group (25 patients) required bolus injection of morphine for postoperative pain relief than those in the fentanyl group (5 patients, P < 0.05). There was no significant difference between the two groups in terms of side effects. CONCLUSIONS: Under the condition of this study protocol, the anesthetic and analgesic effects of remifentanil are more potent than those of fentanyl. Remifentanil can offer superior intraoperative hemodynamic stability in comparison with fentanyl, and has no compromising recovery from anesthesia. PMID- 14527373 TI - Changing constituents of genitourinary cancer in recent 50 years in Beijing. AB - OBJECTIVE: To assess the changes in constituents of genitourinary cancer inpatients in Beijing following the growing economy of China. METHODS: Urological inpatients treated from January 1951 to December 2000 at the Institute of Urology, Peking University were studied retrospectively. RESULTS: In the period of about 50 years 28 474 urological inpatients were reviewed. Cancer patients in 1951-1965 and 1966-2000 comprised 19.5% and 28.2%, respectively [corrected]. Since the 1970s the incidence rates of renal cancer and prostate cancer increased dramatically from 10.4% to 28.7% and from 3.3% to 13.4%, respectively. CONCLUSION: The introduction of new diagnostic instruments, the expansion of the aged population and a high fat diet are considered to be responsible for the change of constituents of genitourinary cancer inpatients. PMID- 14527374 TI - Overexpression of the promyelocytic leukemia gene suppresses growth of human bladder cancer cells by inducing G1 cell cycle arrest and apoptosis. AB - OBJECTIVES: To examine the anti-oncogenic effects of promyelocytic leukemia (PML) on bladder cancer and to explore its molecular mechanisms of growth suppression. METHODS: Wild-type PML was transfected into bladder cancer cells (5637 cell) and expressed in a replication-deficient adenovirus-mediated gene delivery system and introduced into human bladder cancer cells (5637 cell) in vitro and in vivo. The effect and mechanisms of the PML gene in cell growth, clonogenicity, and tumorigenicity of bladder cancer cells were studied using in vitro and in vivo growth assays, soft agar colony-forming assay, cell cycle analysis, apoptosis assay and in vivo tumorigenicity assay. RESULTS: Overexpression of PML in 5637 cells significantly reduced their growth rate and clonogenicity on soft agar. PML suppressed bladder cancer cell growth by inducing G1 cell cycle arrest and apoptosis. Adenovirus-mediated PML (Ad-PML) significantly suppressed the tumorigenicity and growth of bladder cancer cells. Intratumoral injection of Ad PML into tumors induced by 5637 cells dramatically suppressed their growth. CONCLUSIONS: The results indicated that overexpression of PML protein may promote efficient growth inhibition of human bladder cancer cells by inducing G1 cell cycle arrest and apoptosis, and adenovirus-mediated PML (Ad-PML) expression efficiently suppresses human bladder cancer growth. PMID- 14527375 TI - Extracellular matrix synthesis and ultrastructural changes of degenerative disc cells transfected by Ad/CMV-hTGF-beta 1. AB - OBJECTIVE: To determine whether the synthesis of proteoglycan, collagen and associated ultrastructure are related to the adenovirus-mediated gene transferred to adult degenerative cells. METHODS: Adenovirus/cytomegalovirus human transforming growth factor-beta 1 (Ad/CMV-hTGF-beta 1) was used to transfect degenerative cells. Antonopulos method, Miamine method and transmission electron microscopy were conducted to study the synthesis of proteoglycan, collagen, and ultrastructure, respectively. Cell cultures were established from the nucleus pulpous and annulus fibrosus tissues, which were taken from surgery. RESULTS: Nucleus pulpous and annulus fibrosus cells were efficiently transduced by the adenoviral vector carrying hTGF-beta 1 gene. The synthesis of proteoglycan and collagen increased compared with the control group (P < 0.05). The metabolism of cells was slightly improved. No significant toxic effects were found. CONCLUSIONS: Expression of hTGF-beta 1 gene is efficient to accelerates proteoglycan synthesis and thus accelerates the improvement of collagen. The function and structure of degenerative cells are improved. Ad/CMV-hTGF-beta 1 may be suitable for treating disc degeneration. PMID- 14527376 TI - Management of tonsillar squamous cell carcinoma. AB - OBJECTIVE: To discuss treatment options for tonsillar squamous cell carcinoma. METHODS: A total of 108 patients with biopsy-proven tonsillar squamous cell carcinoma, treated between 1984 and 2000, were reviewed, including 82 men and 26 women, with ages ranging from 19 to 70 years. Treatments consisted of either radiotherapy and surgery reserved as salvage treatment (Salvage Surgery, 83 patients), or planned surgery with preoperative radiation (Planned Surgery, 25 patients). Radiotherapy was delivered primarily in a dosage of 60 - 70 Gy for Salvage Surgery patients and 40 - 50 Gy for Planned Surgery patients. Both salvage and planned surgeries were radical, with resection of the lateral oropharyngeal wall, segmental resection of the mandible and neck dissection. The pectoralis major myocutaneous flaps were used to repair surgical defects. RESULTS: The percentages of radical surgery used in the Salvage Surgery and Planned Surgery groups were 24.1% (20/83) and 88.0% (22/25), respectively (P = 0.000). The local recurrence rates were 28.9% (24/83) and 20.0% (5/25) in the Salvage Surgery and Planned Surgery groups, respectively (P = 0.378). The neck recurrence rates were 9.6% (8/83) and 8.0% (2/25) in the Salvage Surgery and Planned Surgery groups respectively (P = 0.804). The 5-year survival rates were 59.3% and 55.3% in the Salvage Surgery and Planned Surgery groups, respectively (P = 0.7056). CONCLUSIONS: Although the two treatments had a similar survival rate, Salvage Surgery avoided 60% commando operations compared with the Planned Surgery group, which benefits to recovery of oral functions. Primary radiotherapy is recommended as the treatment of choice for tonsillar squamous cell carcinoma. After radical radiotherapy, salvage surgery should be undertaken in the case of tumor remnants or recurrences. PMID- 14527377 TI - Expression of C5aR (CD88) of synoviocytes isolated from patients with rheumatoid arthritis and osteoarthritis. AB - OBJECTIVE: To investigate the expression of anaphylatoxin receptor C5aR (CD88) in synoviocytes from patients with rheumatoid arthritis (RA) and osteoarthritis (OA). METHODS: The expression of C5aR was assessed in synoviocytes isolated from 27 RA and 12 OA patients using reverse transcription-polymerase chain reactions (RT-PCR), flow cytometry, and immunofluorescence analysis. The effects of C5a on the release of tumor necrosis factor alpha (TNF alpha) from synoviocytes were assayed using enzyme-linked immunosorbent assays (ELISA). RESULTS: C5aR mRNA was detected in 24 of 27 samples from RA patients, and 10 of 12 samples from OA patients. Flow cytometric analysis and immunofluorescence study demonstrated the cell surface expression of C5aR in a significant proportion of synoviocytes from both RA and OA patients, and the level of C5aR expression in synoviocytes was significantly correlated with joint swelling, erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) in RA patients. Finally, interaction of C5aR with its ligand C5a was shown to enhance lipopolysaccharide (LPS)-induced TNF alpha release from synoviocytes. CONCLUSIONS: The expression of C5aR in synoviocytes from inflammatory joint diseases and also the induction of TNF alpha release in activated synoviocytes by the interaction of C5a and C5aR suggest that the C5a/C5aR system may play an important role in joint inflammation process. PMID- 14527378 TI - Effects of estradiol on proliferation and metabolism of rabbit mandibular condylar cartilage cells in vitro. AB - OBJECTIVE: To investigate the effects in vitro of 17 beta-estradiol (E2) on the proliferation and metabolism of rabbit mandibular condylar cartilage cells. METHODS: Chondrocytes were derived from neonatal rabbit mandibular condylar cartilage using a modified enzyme method. 17 beta-estradiol was added to the culture medium in a variety of concentrations. Cell growth and DNA, collagen, and proteoglycan synthesis were used as indicators of proliferation and differentiation of condylar chondrocytes. These were measured by cell number, 3H proline and 35S-incorporation, respectively. RESULTS: E2 increased cell number and 3H-thymidine incorporation at 10(-8) to 10(-10) mol/L, and 10(-8) to 10(-11) mol/L in a dose-dependent manner, peaking at 10(-8) mol/L and 10(-9) mol/L, respectively. However, further increase in the concentration of estradiol caused inhibition of both cell number and 3H-thymidine incorporation, and this was significant at 10(-6) mol/L. The effect of E2 on proteoglycan synthesis was similar; the maximum stimulating effect was at 10(-8) mol/L, and inhibition was significant at 10(-6) mol/L. There was no obvious stimulatory effect of E2 on 3H thymidine incorporation observed. CONCLUSIONS: Estradiol affects condylar chondrocyte cell growth, DNA, and proteoglycan synthesis in a biphasic manner depending on its concentration. This indicates that estrogen may be important in the proliferation and differentiation of mandibular condylar chondrocytes, and could be relevant to some aspects of certain temporomandibular joint diseases by modulating the function of the chondrocytes. PMID- 14527379 TI - Low dose hormone therapy in reproductive endocrinology in China. PMID- 14527380 TI - Fungal infection in organ transplant patients. AB - PURPOSE: To review the characteristics and evolution of the fungal spectrum, and the risk factors causing fungal infection, and to make progress in diagnosing fungal infection after organ transplantation. DATA SOURCES: An English-language literature search (MEDLINE 1990 - 2000) and bibliographic review of textbooks and review articles. STUDY SELECTION: Twenty-three articles were selected from the literature that specifically addressed the stated purpose. RESULTS: Fungal infections in organ transplant patients were generally divided into two types: (1) disseminated primary or reactivation infection with one of the geographically restricted systemic mycoses; (2) opportunistic infection by fungal species that rarely cause invasive infection in normal hosts. The risk factors of fungal infection after a transplant can be evaluated and predicted according to the organ recipient's conditions before, during and after the transplant. Progress in early diagnostic methods during the past 10 years has mainly revolved around two aspects, culture and non-culture. CONCLUSIONS: It is important to undertake a systemic evaluation on the condition of the organ recipient before, during and after a transplant; should any risk factor for fungal infection be suspected, diagnosis should be made as early as possible by employing mycological techniques including culture and non-culture methods. PMID- 14527381 TI - Effect of continuous positive airway pressure treatment on elderly Chinese patients with obstructive sleep apnea in the prethrombotic state. AB - OBJECTIVES: To characterize the prethrombotic state (PTS) in elderly Chinese patients with obstructive sleep apnea hypopnea syndrome (OSAHS) and the effect of nasal continuous positive airway pressure (nCPAP) ventilation on their PTS. METHODS: Forty-one elderly patients with moderate and severe OSAHS were enrolled into the OSAHS group and underwent nCPAP treatment. Their blood samples were drawn at 6:00 am and 4:00 pm before and during nCPAP treatment, respectively, to test hemocrit, platelet aggregation (PAG), whole blood viscosity (WBV), plasma fibrinogen (fng), prothrombin time (PT) and activated partial thromboplastin time (APTT). All blood factors were also tested in a control group consisting of 32 healthy elderly Chinese with neither OSAHS nor cerebrocardiac vascular disease. RESULTS: In the OSAHS group there was a significantly higher hemocrit, WBV, fng, and a significantly shorter PT and APTT at 6:00 am compared to 4:00 pm before nCPAP treatment, while there was no significant difference among all blood test factors between 6:00 am and 4:00 pm on day 30 of the nCPAP treatment. In the OSAHS group, the hemocrit, WBV, PAG and plasma fng were significantly lower and the PT and APTT were significantly longer at 6:00 am on day 30 of the nCPAP treatment compared to 6:00 am before the nCPAP treatment. A significantly lower hemocrit, but a much longer PT and APTT were observed at 4:00 pm on day 30 of the treatment, compared with 4:00 pm before the treatment. No significant difference among the blood test factors was found between 6:00 am and 4:00 pm blood in the control group or between the control and OSAHS groups after 30 days of nCPAP treatment. CONCLUSION: In elderly Chinese OSAHS patients, PTS could be effectively eliminated by nCPAP treatment. PMID- 14527382 TI - Plants have SOX: the structure of sulfite oxidase from Arabidopsis thaliana. PMID- 14527383 TI - Folding pyrin into the family. PMID- 14527384 TI - Leucine in the sky with diamonds. PMID- 14527385 TI - An atomic model of a plant reovirus: rice dwarf virus. PMID- 14527386 TI - Unraveling the replication machine from negative-stranded RNA viruses. PMID- 14527387 TI - Exploring actin five steps at a time. PMID- 14527388 TI - NMR structure of the apoptosis- and inflammation-related NALP1 pyrin domain. AB - Signaling in apoptosis and inflammation is often mediated by proteins of the death domain superfamily in the Fas/FADD/Caspase-8 or the Apaf-1/Caspase-9 pathways. This superfamily currently comprises the death domain (DD), death effector domain (DED), caspase recruitment domain (CARD), and pyrin domain (PYD) subfamilies. The PYD subfamily is most abundant, but three-dimensional structures are only available for the subfamilies DD, DED, and CARD, which have an antiparallel arrangement of six alpha helices as common fold. This paper presents the NMR structure of PYD of NALP1, a protein that is involved in the innate immune response and is a component of the inflammasome. The structure of NALP1 PYD differs from all other known death domain superfamily structures in that the third alpha helix is replaced by a flexibly disordered loop. This unique feature appears to relate to the molecular basis of familial Mediterranean fever (FMF), a genetic disease caused by single-point mutations. PMID- 14527389 TI - Molecular recognition of paxillin LD motifs by the focal adhesion targeting domain. AB - Focal adhesions (FAs) are large submembrane signaling complexes formed at sites of cellular attachment to the extracellular matrix. The interaction of LD motifs with their targets plays an important role in the assembly of FAs. We have determined the molecular basis for the recognition of two paxillin LD motifs by the FA targeting (FAT) domain of FA kinase using a combination of X-ray crystallography, solution NMR, and homology modeling. The four-helix FAT domain displays two LD binding sites on opposite sites of the molecule that bind LD peptides in a helical conformation. Threading studies suggest that the LD interacting domain of p95PKL shares a common four-helical core with the FAT domain and the tail of vinculin, defining a structural family of LD motif binding modules. PMID- 14527390 TI - Crystal structure of the borna disease virus nucleoprotein. AB - Borna disease virus (BDV) causes an infection of the central nervous system in a wide range of vertebrates, which can fatally progress to an immune-mediated disease, called Borna disease. BDV is a member of the Mononegavirales, which also includes the highly infectious measles and Ebola viruses. The viral nucleoproteins are central to transcription, replication, and packaging of the RNA genome. We present the X-ray structure of the BDV nucleoprotein determined at 1.76 A resolution. The structure reveals a novel fold, organized into two distinct domains, and an assembly into a planar homotetramer. Surface potential calculations strongly support an RNA binding model with the RNA wrapping around the outside of the tetramer, although a positively charged central channel in the tetramer could fit single-stranded RNA in an alternative binding mode. This first structure of an RNA virus nucleoprotein provides a paradigmatic model for RNA packaging and replication of single-stranded RNA viruses. PMID- 14527391 TI - The atomic structure of rice dwarf virus reveals the self-assembly mechanism of component proteins. AB - Rice dwarf virus (RDV), the causal agent of rice dwarf disease, is a member of the genus Phytoreovirus in the family Reoviridae. RDV is a double-shelled virus with a molecular mass of approximately 70 million Dalton. This virus is widely prevalent and is one of the viruses that cause the most economic damage in many Asian countries. The atomic structure of RDV was determined at 3.5 A resolution by X-ray crystallography. The double-shelled structure consists of two different proteins, the core protein P3 and the outer shell protein P8. The atomic structure shows structural and electrostatic complementarities between both homologous (P3-P3 and P8-P8) and heterologous (P3-P8) interactions, as well as overall conformational changes found in P3-P3 dimer caused by the insertion of amino-terminal loop regions of one of the P3 protein into the other. These interactions suggest how the 900 protein components are built into a higher ordered virus core structure. PMID- 14527392 TI - Structural basis for selective recognition of pneumococcal cell wall by modular endolysin from phage Cp-1. AB - Pneumococcal bacteriophage-encoded lysins are modular choline binding proteins that have been shown to act as enzymatic antimicrobial agents (enzybiotics) against streptococcal infections. Here we present the crystal structures of the free and choline bound states of the Cpl-1 lysin, encoded by the pneumococcal phage Cp-1. While the catalytic module displays an irregular (beta/alpha)(5)beta(3) barrel, the cell wall-anchoring module is formed by six similar choline binding repeats (ChBrs), arranged into two different structural regions: a left-handed superhelical domain configuring two choline binding sites, and a beta sheet domain that contributes in bringing together the whole structure. Crystallographic and site-directed mutagenesis studies allow us to propose a general catalytic mechanism for the whole glycoside hydrolase family 25. Our work provides the first complete structure of a member of the large family of choline binding proteins and reveals that ChBrs are versatile elements able to tune the evolution and specificity of the pneumococcal surface proteins. PMID- 14527393 TI - The crystal structure of plant sulfite oxidase provides insights into sulfite oxidation in plants and animals. AB - The molybdenum cofactor (Moco) containing sulfite oxidase (SO) from Arabidopsis thaliana has recently been identified and biochemically characterized. The enzyme is found in peroxisomes and believed to detoxify excess sulfite that is produced during sulfur assimilation, or due to air pollution. Plant SO (PSO) is homodimeric and homologous to animal SO, but contains only a single Moco domain without an additional redox center. Here, we present the first crystal structure of a plant Moco enzyme, the apo-state of Arabidopsis SO at 2.6 A resolution. The overall fold and coordination of the Moco are similar to chicken SO (CSO). Comparisons of conserved surface residues and the charge distribution in PSO and CSO reveal major differences near the entrance to both active sites reflecting different electron acceptors. Arg374 has been identified as an important substrate binding residue due to its conformational change when compared to the sulfate bound structure of CSO. PMID- 14527394 TI - Structural basis for the dual thymidine and thymidylate kinase activity of herpes thymidine kinases. AB - Crystal structures of equine herpesvirus type-4 thymidine kinase (EHV4-TK) in complex with (i). thymidine and ADP, (ii). thymidine and SO(4) and the bisubstrate analogs, (iii). TP(4)A, and (iv). TP(5)A have been solved. Additionally, the structure of herpes simplex virus type-1 thymidine kinase (HSV1 TK) in complex with TP(5)A has been determined. These are the first structures of nucleoside kinases revealing conformational transitions upon binding of bisubstrate analogs. The structural basis for the dual thymidine and thymidylate kinase activity of these TKs is elucidated. While the active sites of HSV1-TK and EHV4-TK resemble one another, notable differences are observed in the Lid regions and in the way the enzymes bind the base of the phosphoryl-acceptor. The latter difference could partly explain the higher activity of EHV4-TK toward the prodrug ganciclovir. PMID- 14527395 TI - Structural plasticity of functional actin: pictures of actin binding protein and polymer interfaces. AB - Actin is one of the most conserved and versatile proteins capable of forming homopolymers and interacting with numerous other proteins in the cell. We performed an alanine mutagenesis scan covering the entire beta-actin molecule. Somewhat surprisingly, the majority of the mutants were capable of reaching a stable conformation. We tested the ability of these mutants to bind to various actin binding proteins, thereby mapping different interfaces with actin. Additionally, we tested their ability to copolymerize with alpha-actin in order to localize regions in actin that contact neighboring protomers in the filament. Hereby, we could discriminate between two existing models for filamentous actin and our data strongly support the right-handed double-stranded helix model. We present data corroborating this model in vivo. Mutants defective in copolymerization do not colocalize with the actin cytoskeleton and some impair its normal function, thereby disturbing cell shape. PMID- 14527396 TI - Structure and interactions of NCAM Ig1-2-3 suggest a novel zipper mechanism for homophilic adhesion. AB - The neural cell adhesion molecule, NCAM, mediates Ca(2+)-independent cell-cell and cell-substratum adhesion via homophilic (NCAM-NCAM) and heterophilic (NCAM non-NCAM molecules) binding. NCAM plays a key role in neural development, regeneration, and synaptic plasticity, including learning and memory consolidation. The crystal structure of a fragment comprising the three N terminal Ig modules of rat NCAM has been determined to 2.0 A resolution. Based on crystallographic data and biological experiments we present a novel model for NCAM homophilic binding. The Ig1 and Ig2 modules mediate dimerization of NCAM molecules situated on the same cell surface (cis interactions), whereas the Ig3 module mediates interactions between NCAM molecules expressed on the surface of opposing cells (trans interactions) through simultaneous binding to the Ig1 and Ig2 modules. This arrangement results in two perpendicular zippers forming a double zipper-like NCAM adhesion complex. PMID- 14527397 TI - A closed compact structure of native Ca(2+)-calmodulin. AB - Calmodulin has been a subject of intense scrutiny since its discovery because of its unusual properties in regulating the functions of about 100 diverse target enzymes and structural proteins. The original and to date only crystal conformation of native eukaryotic Ca(2+)-calmodulin (Ca(2+)-CaM) is a very extended molecule with two widely separated globular domains linked by an exposed long helix. Here we report the 1.7 A X-ray structure of a new native Ca(2+)-CaM that is in a compact ellipsoidal conformation and shows a sharp bend in the linker helix and a more contracted N-terminal domain. This conformation may offer advantages for recognition of kinase-type calmodulin targets or small organic molecule drugs. PMID- 14527398 TI - Synaptonemal complexities and commonalities. PMID- 14527399 TI - Soaking up RNAi. PMID- 14527400 TI - Zero tolerance for nonsense: nonsense-mediated mRNA decay uses multiple degradation pathways. PMID- 14527401 TI - Keeping RNA and DNA apart during transcription. PMID- 14527402 TI - An open-and-shut case? Recent insights into the activation of EGF/ErbB receptors. AB - Recent crystallographic studies have provided significant new insight into how receptor tyrosine kinases from the EGF receptor or ErbB family are regulated by their growth factor ligands. EGF receptor dimerization is mediated by a unique dimerization arm, which becomes exposed only after a dramatic domain rearrangement is promoted by growth factor binding. ErbB2, a family member that has no ligand, has its dimerization arm constitutively exposed, and this explains several of its unique properties. We outline a mechanistic view of ErbB receptor homo- and heterodimerization, which suggests new approaches for interfering with these processes when they are implicated in human cancers. PMID- 14527403 TI - Essential role of E2-25K/Hip-2 in mediating amyloid-beta neurotoxicity. AB - The ubiquitin/proteasome system has been proposed to play an important role in Alzheimer's disease (AD) pathogenesis. However, the critical factor(s) modulating both amyloid-beta peptide (Abeta) neurotoxicity and ubiquitin/proteasome system in AD are not known. We report the isolation of an unusual ubiquitin-conjugating enzyme, E2-25K/Hip-2, as a mediator of Abeta toxicity. The expression of E2 25K/Hip-2 was upregulated in the neurons exposed to Abeta(1-42) in vivo and in culture. Enzymatic activity of E2-25K/Hip-2 was required for both Abeta(1-42) neurotoxicity and inhibition of proteasome activity. E2-25K/Hip-2 functioned upstream of apoptosis signal-regulating kinase 1 (ASK1) and c-Jun N-terminal kinase (JNK) in Abeta(1-42) toxicity. Further, the ubiquitin mutant, UBB+1, a potent inhibitor of the proteasome which is found in Alzheimer's brains, was colocalized and functionally interacted with E2-25K/Hip-2 in mediating neurotoxicity. These results suggest that E2-25K/Hip-2 is a crucial factor in regulating Abeta neurotoxicity and could play a role in the pathogenesis of Alzheimer's disease. PMID- 14527404 TI - The group II chaperonin TRiC protects proteolytic intermediates from degradation in the MHC class I antigen processing pathway. AB - MHC class I molecules present precisely cleaved peptides of intracellular proteins on the cell surface. For most antigenic precursors, presentation requires transport of peptide fragments into the ER, but the nature of the cytoplasmic peptides and their chaperones is obscure. By tracking proteolytic intermediates in living cells, we show that intracellular proteolysis yields a mixture of antigenic peptides containing only N-terminal flanking residues for ER transport. Some of these peptides were bound to the group II chaperonin TRiC and were protected from degradation. Destabilization of TRiC by RNA interference inhibited the expression of peptide-loaded MHC I molecules on the cell surface. Thus, the TRiC chaperonin serves a function in protecting proteolytic intermediates in the MHC I antigen processing pathway. PMID- 14527405 TI - Convergent mechanisms for recognition of divergent cytokines by the shared signaling receptor gp130. AB - Gp130 is a shared cell-surface signaling receptor for at least ten different hematopoietic cytokines, but the basis of its degenerate recognition properties is unknown. We have determined the crystal structure of human leukemia inhibitory factor (LIF) bound to the cytokine binding region (CHR) of gp130 at 2.5 A resolution. Strikingly, we find that the shared binding site on gp130 has an entirely rigid core, while the LIF binding interface diverges sharply in structure and chemistry from that of other gp130 ligands. Dissection of the LIF gp130 interface, along with comparative studies of other gp130 cytokines, reveal that gp130 has evolved a "thermodynamic plasticity" that is relatively insensitive to ligand structure, to enable crossreactivity. These observations reveal a novel and alternative mechanism for degenerate recognition from that of structural plasticity. PMID- 14527406 TI - HIV-1 Vif blocks the antiviral activity of APOBEC3G by impairing both its translation and intracellular stability. AB - The human immunodeficiency virus type 1 (HIV-1) relies on Vif (viral infectivity factor) to overcome the potent antiviral function of APOBEC3G (apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like 3G, also known as CEM15). Using an APOBEC3G-specific antiserum, we now show that Vif prevents virion incorporation of endogenous APOBEC3G by effectively depleting the intracellular levels of this enzyme in HIV-1-infected T cells. Vif achieves this depletion by both impairing the translation of APOBEC3G mRNA and accelerating the posttranslational degradation of the APOBEC3G protein by the 26S proteasome. Vif physically interacts with APOBEC3G, and expression of Vif alone in the absence of other HIV-1 proteins is sufficient to cause depletion of APOBEC3G. These findings highlight how the bimodal translational and posttranslational inhibitory effects of Vif on APOBEC3G combine to markedly suppress the expression of this potent antiviral enzyme in virally infected cells, thereby effectively curtailing the incorporation of APOBEC3G into newly formed HIV-1 virions. PMID- 14527407 TI - Retroviral insertional mutagenesis identifies a small protein required for synthesis of diphthamide, the target of bacterial ADP-ribosylating toxins. AB - Retroviral insertional mutagenesis was used to produce a mutant Chinese hamster ovary cell line that is completely resistant to several different bacterial ADP ribosylating toxins. The gene responsible for toxin resistance, termed diphtheria toxin (DT) and Pseudomonas exotoxin A (ETA) sensitivity required gene 1 (DESR1), encodes two small protein isoforms of 82 and 57 residues. DESR1 is evolutionally conserved and ubiquitously expressed. Only the longer isoform is functional because the mutant cell line can be complemented by transfection with the long but not the short isoform. We demonstrate that DESR1 is required for the first step in the posttranslational modification of elongation factor-2 at His(715) that yields diphthamide, the target site for ADP ribosylation by DT and ETA. KTI11, the analog of DESR1 in yeast, which was originally identified as a gene regulating the sensitivity of yeast to zymocin, is also required for diphthamide biosynthesis, implicating DESR1/KTI11 in multiple biological processes. PMID- 14527408 TI - Conserved structural motifs in intracellular trafficking pathways: structure of the gammaCOP appendage domain. AB - The formation of coated vesicles is a fundamental step in many intracellular trafficking pathways. COPI and clathrin represent two important and distinct sets of vesicle coating machinery, involved primarily in mediating intra-Golgi and endocytic transport, respectively. Here we identify an important functional region at the carboxyl terminus of the gamma subunit of the COPI complex (gammaCOP) and describe the X-ray crystal structure of this domain at 2.3 A resolution. This domain of gammaCOP exhibits unexpected structural similarity to the carboxyl-terminal appendage domains of the alpha and beta subunits of the AP2 adaptor proteins, integral components of clathrin-coated vesicles. The remarkable structural conservation exhibited by the gammaCOP appendage domain, coupled with functional data and primary sequence analysis, supports a model of COPI function with significant structural and mechanistic parallels to vesicular transport by the clathrin/AP2 system. PMID- 14527409 TI - Identification of modulators of TRAIL-induced apoptosis via RNAi-based phenotypic screening. AB - New opportunities in mammalian functional genomics are emerging through the combination of high throughput technology and methods that allow manipulation of gene expression in living cells. Here we describe the application of an RNAi based forward genomics approach toward understanding the biology and mechanism of TRAIL-induced apoptosis. TRAIL is a TNF superfamily member that induces selective cytotoxicity of tumor cells when bound to its cognate receptors. In addition to detecting well-characterized genes in the apoptosis pathway, we uncover several modulators including DOBI, a gene required for progression of the apoptotic signal through the intrinsic mitochondrial cell death pathway, and MIRSA, a gene that acts to limit TRAIL-induced apoptosis. Moreover, our data suggest a role for MYC and the WNT pathway in maintaining susceptibility to TRAIL. Collectively, these observations offer several insights on how TRAIL mediates the selective killing of tumor cells and demonstrate the utility of large-scale RNAi screens in mammalian cells. PMID- 14527410 TI - pRB contains an E2F1-specific binding domain that allows E2F1-induced apoptosis to be regulated separately from other E2F activities. AB - The interaction between pRB and E2F is critical for control of the cell cycle and apoptosis. Here we report that pRB contains two distinct E2F binding sites. The previously identified E2F binding site on pRB is necessary for stable association with E2Fs on DNA. A second E2F interaction site is located entirely within the C terminal domain of pRB and is specific for E2F1. E2F1/pRB complexes formed through this site have low affinity for DNA, but the interaction is sufficient for pRB to regulate E2F1-induced apoptosis, and E2F1 loses the ability to interact with this site following DNA damage. These results show that pRB interacts with individual E2F proteins in different ways and suggest that pRB's regulation of E2F1-induced apoptosis is physically separable from its transcriptional control of other E2F proteins. PMID- 14527411 TI - A tweezers-like motion of the ATP-binding cassette dimer in an ABC transport cycle. AB - The ATPase components of ATP binding cassette (ABC) transporters power the transporters by binding and hydrolyzing ATP. Major conformational changes of an ATPase are revealed by crystal structures of MalK, the ATPase subunit of the maltose transporter from Escherichia coli, in three different dimeric configurations. While other nucleotide binding domains or subunits display low affinity for each other in the absence of the transmembrane segments, the MalK dimer is stabilized through interactions of the additional C-terminal domains. In the two nucleotide-free structures, the N-terminal nucleotide binding domains are separated to differing degrees, and the dimer is maintained through contacts of the C-terminal regulatory domains. In the ATP-bound form, the nucleotide binding domains make contact and two ATPs lie buried along the dimer interface. The two nucleotide binding domains of the dimer open and close like a pair of tweezers, suggesting a regulatory mechanism for ATPase activity that may be tightly coupled to translocation. PMID- 14527412 TI - Cytoskeletal activation of a checkpoint kinase. AB - The assembly of cytoskeletal structures is coupled to other cellular processes. We have studied the molecular mechanism by which assembly of the yeast septin cytoskeleton is monitored and coordinated with cell cycle progression by analyzing a key regulatory protein kinase, Hsl1, that becomes activated only when the septin cytoskeleton is properly assembled. We first identified a regulatory region of Hsl1 that physically associates with the kinase domain and found that it performs an autoinhibitory function both in vivo and in vitro. Several septin binding domains lie near and overlap the inhibitory domain; these are important for Hsl1 function, and binding of two septins, Cdc11 and Cdc12, relieves the autoinhibition imposed by the kinase inhibitory domain in vitro. Our results suggest that binding to multiple septins activates Hsl1 kinase activity, thereby promoting cell cycle progression. The high conservation of Hsl1 indicates that similar mechanisms may monitor cytoskeletal organization in other eukaryotes. PMID- 14527413 TI - Nonsense-mediated mRNA decay in mammalian cells involves decapping, deadenylating, and exonucleolytic activities. AB - Nonsense-mediated mRNA decay (NMD) is a mechanism by which cells recognize and degrade mRNAs that prematurely terminate translation. To date, the polarity and enzymology of NMD in mammalian cells is unknown. We show here that downregulating the Dcp2 decapping protein or the PM/Scl100 component of the exosome (1) significantly increases the abundance of steady-state nonsense-containing but not nonsense-free mRNAs, and (2) significantly slows the decay rate of transiently induced nonsense-containing but not nonsense-free mRNA. Downregulating poly(A) ribonuclease (PARN) also increases the abundance of nonsense-containing mRNAs. Furthermore, NMD factors Upf1, Upf2, and Upf3X coimmunopurify with the decapping enzyme Dcp2, the putative 5'-->3' exonuclease Rat1, the proven 5'-->3' exonuclease Xrn1, exosomal components PM/Scl100, Rrp4, and Rrp41, and PARN. From these and other data, we conclude that NMD in mammalian cells degrades mRNAs from both 5' and 3' ends by recruiting decapping and 5'-->3' exonuclease activities as well as deadenylating and 3'-->5' exonuclease activities. PMID- 14527414 TI - An RNA-modifying enzyme that governs both the codon and amino acid specificities of isoleucine tRNA. AB - The AUA codon-specific isoleucine tRNA (tRNA(Ile)) in eubacteria has the posttranscriptionally modified nucleoside lysidine (L) at the wobble position of the anticodon (position 34). This modification is a lysine-containing cytidine derivative that converts both the codon specificity of tRNA(Ile) from AUG to AUA and its amino acid specificity from methionine to isoleucine. We identified an essential gene (tilS; tRNA(Ile)-lysidine synthetase) that is responsible for lysidine formation in both Bacillus subtilis and Escherichia coli. The recombinant enzyme complexed specifically with tRNA(Ile) and synthesized L by utilizing ATP and lysine as substrates. The lysidine synthesis of this enzyme was shown to directly convert the amino acid specificity of tRNA(Ile) from methionine to isoleucine in vitro. Partial inactivation of tilS in vivo resulted in an AUA codon-dependent translational defect, which supports the notion that TilS is an RNA-modifying enzyme that plays a critical role in the accurate decoding of genetic information. PMID- 14527415 TI - A minimal RNA polymerase III transcription system from human cells reveals positive and negative regulatory roles for CK2. AB - In higher eukaryotes, RNA polymerase (pol) III is known to use different transcription factors to recognize three basic types of promoters, but in no case have these transcription factors been completely defined. We show that a highly purified pol III complex combined with the recombinant transcription factors SNAP(c), TBP, Brf2, and Bdp1 directs multiple rounds of transcription initiation and termination from the human U6 promoter. The pol III complex contains traces of CK2, and CK2 associates with the U6 promoter region in vivo. Transcription requires CK2 phosphorylation of the pol III complex. In contrast, CK2 phosphorylation of TBP, Brf2, and Bdp1 combined is inhibitory. The results define a minimum core machinery, the ultimate target of regulatory mechanisms, capable of directing all steps of the transcription process-initiation, elongation, and termination-by a metazoan RNA polymerase, and suggest positive and negative regulatory roles for CK2 in transcription by pol III. PMID- 14527416 TI - Cotranscriptionally formed DNA:RNA hybrids mediate transcription elongation impairment and transcription-associated recombination. AB - Genetic instability, a phenomenon relevant for developmentally regulated processes, cancer, and inherited disorders, can be induced by transcription. However, the mechanisms of transcription-associated genetic instability are not yet understood. Analysis of S. cerevisiae mutants of THO/TREX, a conserved eukaryotic protein complex functioning at the interface of transcription and mRNA metabolism, has provided evidence that transcription elongation impairment can cause hyperrecombination. Here we show, using hpr1Delta mutants, that the nascent mRNA can diminish transcription elongation efficiency and promote recombination. If during transcription the nascent mRNA is self-cleaved by a hammerhead ribozyme, the transcription-defect and hyperrecombination phenotypes of hpr1Delta cells are suppressed. Abolishment of hyperrecombination by overexpression of RNase H1 and molecular detection of DNA:RNA hybrids indicate that these are formed cotranscriptionally in hpr1Delta cells. These data support a model to explain the connection between recombination, transcription, and mRNA metabolism and provide a new perspective to understanding transcription-associated recombination. PMID- 14527417 TI - N-CoR mediates DNA methylation-dependent repression through a methyl CpG binding protein Kaiso. AB - The identification and characterization of molecular mechanisms utilized by cells to mediate transcriptional repression at methylated loci are fundamental to understanding the biological consequences of DNA methylation. Here we demonstrate that Kaiso, a methyl CpG binding protein belonging to the BTB/POZ family of transcription factors, is a component of the human N-CoR complex. In vitro, the Kaiso/N-CoR complex binds specific CpG-rich sequences in a methylation-dependent manner. In vivo, Kaiso targets the N-CoR complex to the MTA2 gene promoter in a methylation-dependent manner. Importantly, we demonstrate that Kaiso is required for transcriptional repression of the methylated MTA2 locus. Furthermore, this repression also requires a functional N-CoR deacetylase complex, which brings about histone hypoacetylation and methylation of H3 lysine 9 to the MTA2 locus. Thus, our data demonstrate a critical role for a methyl CpG binding protein in mediating DNA methylation-dependent repression and reveal the mechanism by which it represses transcription. PMID- 14527418 TI - Active localization of the retinoblastoma protein in chromatin and its response to S phase DNA damage. AB - The Rb protein suppresses development of an abnormal state of endoreduplication arising after S phase DNA damage. In diploid, S phase cells, the activity of protein phosphatase 2A (PP2A) licenses the stable association of un(der)phosphorylated Rb with chromatin. After damage, chromatin-associated pRb is attracted to certain chromosomal replication initiation sites in the order in which they normally fire. Like S phase DNA damage in Rb(-/-) cells, specific interruption of PP2A function in irradiated, S phase wt cells also elicited a state of endoreduplication. Thus, PP2A normally licenses the recruitment of Rb to chromatin sites in S phase from which, after DNA damage, it relocalizes to selected replication control sites and suppresses abnormal, postdamage rereplicative activity. PMID- 14527419 TI - The endogenous Mus81-Eme1 complex resolves Holliday junctions by a nick and counternick mechanism. AB - Functional studies strongly suggest that the Mus81-Eme1 complex resolves Holliday junctions (HJs) in fission yeast, but in vitro it preferentially cleaves flexible three-way branched structures that model replication forks or 3' flaps. Here we report that a nicked HJ is the preferred substrate of endogenous and recombinant Mus81-Eme1. Cleavage occurs specifically on the strand that opposes the nick, resulting in resolution of the structure into linear duplex products. Resolving cuts made by the endogenous Mus81-Eme1 complex on an intact HJ are quasi simultaneous, indicating that Mus81-Eme1 resolves HJs by a nick and counternick mechanism, with a large rate enhancement of the second cut arising from the flexible nature of the nicked HJ intermediate. Recombinant Mus81-Eme1 is ineffective at making the first cut. We also report that HJs accumulate in a DNA polymerase alpha mutant that lacks Mus81, providing further evidence that the Mus81-Eme1 complex targets HJs in vivo. PMID- 14527420 TI - Generating crossovers by resolution of nicked Holliday junctions: a role for Mus81-Eme1 in meiosis. AB - The double Holliday junction (dHJ) is generally regarded to be a key intermediate of meiotic recombination, whose resolution is critical for the formation of crossover recombinants. In fission yeast, the Mus81-Eme1 endonuclease has been implicated in resolving dHJs. Consistent with this role, we show that Mus81-Eme1 is required for generating meiotic crossovers. However, purified Mus81-Eme1 prefers to cleave junctions that mimic those formed during the transition from double-strand break to dHJ. Crucially, these junctions are cleaved by Mus81-Eme1 in precisely the right orientation to guarantee the formation of a crossover every time. These data demonstrate how crossovers could arise without forming or resolving dHJs using an enzyme that is widely conserved amongst eukaryotes. PMID- 14527421 TI - The architecture of the gammadelta resolvase crossover site synaptic complex revealed by using constrained DNA substrates. AB - Activated mutants of the serine recombinase, gammadelta resolvase, form a simplified recombinogenic synaptic complex containing a tetramer of resolvase and two crossover sites. We have probed the architecture of this complex by measuring the efficiency of recombination of a series of constrained DNA substrates (with phased recombination sites separated by an IHF-induced U-turn); this serves as a direct report on the topology of a productive synapse. Our data show that in the active complex, the catalytic domains from two resolvase dimers form a central core, while the DNA binding domains and the DNA lie on the outside. In addition, the crossover sites cross one another to form a local positive node. The implications of our data for the mechanism of strand exchange and the process of resolvase activation are discussed. PMID- 14527422 TI - BTB/POZ domain proteins are putative substrate adaptors for cullin 3 ubiquitin ligases. AB - Cullins (CULs) are subunits of a prominent class of RING ubiquitin ligases. Whereas the subunits and substrates of CUL1-associated SCF complexes and CUL2 ubiquitin ligases are well established, they are largely unknown for other cullin family members. We show here that S. pombe CUL3 (Pcu3p) forms a complex with the RING protein Pip1p and all three BTB/POZ domain proteins encoded in the fission yeast genome. The integrity of the BTB/POZ domain, which shows similarity to the cullin binding proteins SKP1 and elongin C, is required for this interaction. Whereas Btb1p and Btb2p are stable proteins, Btb3p is ubiquitylated and degraded in a Pcu3p-dependent manner. Btb3p degradation requires its binding to a conserved N-terminal region of Pcu3p that precisely maps to the equivalent SKP1/F box adaptor binding domain of CUL1. We propose that the BTB/POZ domain defines a recognition motif for the assembly of substrate-specific RING/cullin 3/BTB ubiquitin ligase complexes. PMID- 14527425 TI - Polyglutamines stop traffic: axonal transport as a common target in neurodegenerative diseases. AB - In this issue of Neuron, two papers provide evidence that polyglutamine (polyQ) proteins disrupt axonal transport. Gunawardena et al. show that normal levels of huntingtin are required for proper axonal transport in Drosophila. In addition, expression of expanded polyQ proteins disrupts axonal transport in larval neurons. Szebenyi and colleagues find that polyQ proteins directly inhibit fast axonal transport using axoplasm from the squid giant axon and suggest that axonal transport defects may be a common feature of polyQ disease pathogenesis. PMID- 14527426 TI - Intracellular Ca2+ release and ischemic axon injury: the Trojan horse is back. AB - Ischemic injury of cells in the central nervous system is typically set in motion by influx of extracellular Ca(2+). In this issue of Neuron, Stys and colleagues propose that ischemic injury in spinal cord axons is partly the result of ryanodine receptor-mediated release of Ca(2+) from the endoplasmic reticulum (ER), a site of intracellular Ca(2+) storage. PMID- 14527427 TI - Downstream of guidance receptors: entering the baroque period of axon guidance signaling. AB - The Roundabout ("Robo") family of transmembrane proteins are the receptors and mediators of the repellent axon guidance signal Slit. However, the molecular mechanisms by which Robo signaling leads to growth cone or neuron repulsion are still poorly understood. A study by Fan et al. in this issue of Neuron expands the repertoire of Robo pathway components and stimulates a new look at axon guidance signaling in general. PMID- 14527428 TI - How to make a relationship last: release sites with different levels of commitment. AB - Stimulus-induced changes in synaptic strength endure for periods ranging from minutes to days. In this issue of Neuron, Kim et al. show that two distinct types of synaptic change-synaptic vesicle movement into previously empty varicosities and formation of new varicosities-may determine intermediate- versus long-term synaptic facilitation. PMID- 14527429 TI - Somitogenesis: breaking new boundaries. AB - Segmentation is a fundamental process in vertebrate embryogenesis, and one of the earliest manifestations of segmental patterning is the generation of transient, serially repeated blocks of mesodermal cells known as somites. Disruption of the normal segmentation process in humans leads to vertebral abnormalities such as spondylocostal dysostosis. In this minireview, we discuss recent advances in the dynamic molecular and cellular mechanisms governing segmentation. PMID- 14527430 TI - Charybdotoxin binding in the I(Ks) pore demonstrates two MinK subunits in each channel complex. AB - I(Ks) voltage-gated K(+) channels contain four pore-forming KCNQ1 subunits and MinK accessory subunits in a number that has been controversial. Here, I(Ks) channels assembled naturally by monomer subunits are compared to those with linked subunits that force defined stoichiometries. Two strategies that exploit charybdotoxin (CTX)-sensitive subunit variants are applied. First, CTX on rate, off rate, and equilibrium affinity are found to be the same for channels of monomers and those with a fixed 2:4 MinK:KCNQ1 valence. Second, 3H-CTX and an antibody are used to directly quantify channels and MinK subunits, respectively, showing 1.97 +/- 0.07 MinK per I(Ks) channel. Additional MinK subunits do not enter channels of monomeric subunits or those with fixed 2:4 valence. We conclude that two MinK subunits are necessary, sufficient, and the norm in I(Ks) channels. This stoichiometry is expected for other K(+) channels that contain MinK or MinK related peptides (MiRPs). PMID- 14527431 TI - Disruption of axonal transport by loss of huntingtin or expression of pathogenic polyQ proteins in Drosophila. AB - We tested whether proteins implicated in Huntington's and other polyglutamine (polyQ) expansion diseases can cause axonal transport defects. Reduction of Drosophila huntingtin and expression of proteins containing pathogenic polyQ repeats disrupt axonal transport. Pathogenic polyQ proteins accumulate in axonal and nuclear inclusions, titrate soluble motor proteins, and cause neuronal apoptosis and organismal death. Expression of a cytoplasmic polyQ repeat protein causes adult retinal degeneration, axonal blockages in larval neurons, and larval lethality, but not neuronal apoptosis or nuclear inclusions. A nuclear polyQ repeat protein induces neuronal apoptosis and larval lethality but no axonal blockages. We suggest that pathogenic polyQ proteins cause neuronal dysfunction and organismal death by two non-mutually exclusive mechanisms. One mechanism requires nuclear accumulation and induces apoptosis; the other interferes with axonal transport. Thus, disruption of axonal transport by pathogenic polyQ proteins could contribute to early neuropathology in Huntington's and other polyQ expansion diseases. PMID- 14527432 TI - Neuropathogenic forms of huntingtin and androgen receptor inhibit fast axonal transport. AB - Huntington's and Kennedy's disease are autosomal dominant neurodegenerative diseases caused by pathogenic expansion of polyglutamine tracts. Expansion of glutamine repeats must in some way confer a gain of pathological function that disrupts an essential cellular process and leads to loss of affected neurons. Association of huntingtin with vesicular structures raised the possibility that axonal transport might be altered. Here we show that polypeptides containing expanded polyglutamine tracts, but not normal N-terminal huntingtin or androgen receptor, directly inhibit both fast axonal transport in isolated axoplasm and elongation of neuritic processes in intact cells. Effects were greater with truncated polypeptides and occurred without detectable morphological aggregates. PMID- 14527433 TI - Depolarization-induced Ca2+ release in ischemic spinal cord white matter involves L-type Ca2+ channel activation of ryanodine receptors. AB - The mechanisms of Ca(2+) release from intracellular stores in CNS white matter remain undefined. In rat dorsal columns, electrophysiological recordings showed that in vitro ischemia caused severe injury, which persisted after removal of extracellular Ca(2+); Ca(2+) imaging confirmed that an axoplasmic Ca(2+) rise persisted in Ca(2+)-free perfusate. However, depletion of Ca(2+) stores or reduction of ischemic depolarization (low Na(+), TTX) were protective, but only in Ca(2+)-free bath. Ryanodine or blockers of L-type Ca(2+) channel voltage sensors (nimodipine, diltiazem, but not Cd(2+)) were also protective in zero Ca(2+), but their effects were not additive with ryanodine. Immunoprecipitation revealed an association between L-type Ca(2+) channels and RyRs, and immunohistochemistry confirmed colocalization of Ca(2+) channels and RyR clusters on axons. Similar to "excitation-contraction coupling" in skeletal muscle, these results indicate a functional coupling whereby depolarization sensed by L-type Ca(2+) channels activates RyRs, thus releasing damaging amounts of Ca(2+) under pathological conditions in white matter. PMID- 14527434 TI - Opposing FGF and retinoid pathways control ventral neural pattern, neuronal differentiation, and segmentation during body axis extension. AB - Vertebrate body axis extension involves progressive generation and subsequent differentiation of new cells derived from a caudal stem zone; however, molecular mechanisms that preserve caudal progenitors and coordinate differentiation are poorly understood. FGF maintains caudal progenitors and its attenuation is required for neuronal and mesodermal differentiation and to position segment boundaries. Furthermore, somitic mesoderm promotes neuronal differentiation in part by downregulating Fgf8. Here we identify retinoic acid (RA) as this somitic signal and show that retinoid and FGF pathways have opposing actions. FGF is a general repressor of differentiation, including ventral neural patterning, while RA attenuates Fgf8 in neuroepithelium and paraxial mesoderm, where it controls somite boundary position. RA is further required for neuronal differentiation and expression of key ventral neural patterning genes. Our data demonstrate that FGF and RA pathways are mutually inhibitory and suggest that their opposing actions provide a global mechanism that controls differentiation during axis extension. PMID- 14527435 TI - A requirement for retinoic acid-mediated transcriptional activation in ventral neural patterning and motor neuron specification. AB - The specification of neuronal fates in the ventral spinal cord depends on the regulation of homeodomain (HD) and basic-helix-loop-helix (bHLH) proteins by Sonic hedgehog (Shh). Most of these transcription factors function as repressors, leaving unresolved the link between inductive signaling pathways and transcriptional activators involved in ventral neuronal specification. We show here that retinoid signaling and the activator functions of retinoid receptors are required to pattern the expression of HD and bHLH proteins and to specify motor neuron identity. We also show that fibroblast growth factors (FGFs) repress progenitor HD protein expression, implying that evasion of FGF signaling and exposure to retinoid and Shh signals are obligate steps in the emergence of ventral neural pattern. Moreover, joint exposure of neural progenitors to retinoids and FGFs suffices to induce motor neuron differentiation in a Shh independent manner. PMID- 14527436 TI - Retinoid receptor signaling in postmitotic motor neurons regulates rostrocaudal positional identity and axonal projection pattern. AB - The identity of motor neurons diverges markedly at different rostrocaudal levels of the spinal cord, but the signals that specify their fate remain poorly defined. We show that retinoid receptor activation in newly generated spinal motor neurons has a crucial role in specifying motor neuron columnar subtypes. Blockade of retinoid receptor signaling in brachial motor neurons inhibits lateral motor column differentiation and converts many of these neurons to thoracic columnar subtypes. Conversely, expression of a constitutively active retinoid receptor derivative impairs the differentiation of thoracic motor neuron columnar subtypes. These findings provide evidence for a regionally restricted role for retinoid signaling in the postmitotic specification of motor neuron columnar identity. PMID- 14527437 TI - Slit stimulation recruits Dock and Pak to the roundabout receptor and increases Rac activity to regulate axon repulsion at the CNS midline. AB - Drosophila Roundabout (Robo) is the founding member of a conserved family of repulsive axon guidance receptors that respond to secreted Slit proteins. Here we present evidence that the SH3-SH2 adaptor protein Dreadlocks (Dock), the p21 activated serine-threonine kinase (Pak), and the Rac1/Rac2/Mtl small GTPases can function during Robo repulsion. Loss-of-function and genetic interaction experiments suggest that limiting the function of Dock, Pak, or Rac partially disrupts Robo repulsion. In addition, Dock can directly bind to Robo's cytoplasmic domain, and the association of Dock and Robo is enhanced by stimulation with Slit. Furthermore, Slit stimulation can recruit a complex of Dock and Pak to the Robo receptor and trigger an increase in Rac1 activity. These results provide a direct physical link between the Robo receptor and an important cytoskeletal regulatory protein complex and suggest that Rac can function in both attractive and repulsive axon guidance. PMID- 14527438 TI - Carbon monoxide neurotransmission activated by CK2 phosphorylation of heme oxygenase-2. AB - Carbon monoxide (CO) is a putative gaseous neurotransmitter that lacks vesicular storage and must be synthesized rapidly following neuronal depolarization. We show that the biosynthetic enzyme for CO, heme oxygenase-2 (HO2), is activated during neuronal stimulation by phosphorylation by CK2 (formerly casein kinase 2). Phorbol ester treatment of hippocampal cultures results in the phosphorylation and activation of HO2 by CK2, implicating protein kinase C (PKC) in CK2 stimulation. Odorant treatment of olfactory receptor neurons augments HO2 phosphorylation and activity as well as cyclic guanosine monophosphate (cGMP) levels, with all of these effects selectively blocked by CK2 inhibitors. Likewise, CO-mediated nonadrenergic, noncholinergic (NANC) relaxation of the internal anal sphincter requires CK2 activity. Our findings provide a molecular mechanism for the rapid neuronal activation of CO biosynthesis, as required for a gaseous neurotransmitter. PMID- 14527439 TI - Hypocretin (orexin) induces calcium transients in single spines postsynaptic to identified thalamocortical boutons in prefrontal slice. AB - In vivo, thalamocortical axons are susceptible to the generation of terminal spikes which antidromically promote bursting in the thalamus. Although neurotransmitters could elicit such ectopic action potentials at thalamocortical boutons, this hypothesis has never been confirmed. Prefrontal cortex is the cortical area most implicated in arousal and is innervated by thalamic neurons that are unusual since they burst rhythmically during waking. We show that a neurotransmitter critical for alertness, hypocretin (orexin), directly excites prefrontal thalamocortical synapses in acute slice. This TTX-sensitive activation of thalamic axons was demonstrated electrophysiologically and by two-photon sampling of calcium transients at single spines in apposition to thalamic boutons anterogradely labeled in vivo. Spines receiving these long-range projections constituted a unique population in terms of the presynaptic excitatory action of hypocretin. By this mechanism, the hypocretin projection to prefrontal cortex may play a larger role in prefrontal or "executive" aspects of alertness and attention than previously anticipated. PMID- 14527440 TI - Presynaptic activation of silent synapses and growth of new synapses contribute to intermediate and long-term facilitation in Aplysia. AB - The time course and functional significance of the structural changes associated with long-term facilitation of Aplysia sensory to motor neuron synaptic connections in culture were examined by time-lapse confocal imaging of individual sensory neuron varicosities labeled with three different fluorescent markers: the whole-cell marker Alexa-594 and two presynaptic marker proteins-synaptophysin eGFP to monitor changes in synaptic vesicle distribution and synapto-PHluorin to monitor active transmitter release sites. Repeated pulses of serotonin induce two temporally, morphologically, and molecularly distinct presynaptic changes: (1) a rapid activation of silent presynaptic terminals by filling of preexisting empty varicosities with synaptic vesicles, which parallels intermediate-term facilitation, is completed within 3-6 hr and requires translation but not transcription and (2) a slower generation of new functional varicosities which occurs between 12-18 hr and requires transcription and translation. Enrichment of empty varicosities with synaptophysin accounts for 32% of the newly activated synapses at 24 hr, whereas newly formed varicosities account for 68%. PMID- 14527444 TI - pH dependence of uranyl retention in a quartz/solution system: an XPS study. AB - We have investigated the pH dependence of U(VI) retention in quartz/10(-4) M uranyl solution systems, under conditions favoring formation of polynuclear aqueous species and of colloids of amorphous schoepite as U(VI) solubility limiting phases. X-ray photoelectron spectroscopy was used to gain insights into the coordination environments of sorbed/precipitated uranyl ions in the centrifuged quartz samples. The U4f XPS spectra made it possible to identify unambiguously the presence of two uranyl components. A high binding energy component, whose relative proportion increases with pH, exhibits the U4f lines characteristic of a reference synthetic metaschoepite. Such a high binding energy component is interpreted as a component having a U(VI) oxide hydrate character, either as polynuclear surface oligomers and/or as amorphous schoepite-like (surface) precipitates. Its pH dependence suggests that a binding of polynuclear species at quartz surfaces and/or a formation of amorphous schoepite-like (surface) precipitates is favored when the proportion of aqueous polynuclear species increases. A second surface component exhibits binding energies for the U4f core levels at values significantly lower (DeltaE(b)=1.2 eV) than for metaschoepite, evidencing uranyl ions in a distinct coordination environment. Such a low binding energy component may be attributed to monomeric uranyl surface complexes on the basis of published EXAFS data. Such a hypothesis is supported by a major contribution of the low binding energy component to the U4f XPS spectra of reference samples for uranyl sorbed on quartz from very acidic 10(-3) M uranyl solutions where UO(2)(2+) ions predominate. PMID- 14527442 TI - Dynamics of population code for working memory in the prefrontal cortex. AB - Some neurons (delay cells) in the prefrontal cortex elevate their activities throughout the time period during which the animal is required to remember past events and prepare future behavior, suggesting that working memory is mediated by continuous neural activity. It is unknown, however, how working memory is represented within a population of prefrontal cortical neurons. We recorded from neuronal ensembles in the prefrontal cortex as rats learned a new delayed alternation task. Ensemble activities changed in parallel with behavioral learning so that they increasingly allowed correct decoding of previous and future goal choices. In well-trained rats, considerable decoding was possible based on only a few neurons and after removing continuously active delay cells. These results show that neural activity in the prefrontal cortex changes dynamically during new task learning so that working memory is robustly represented and that working memory can be mediated by sequential activation of different neural populations. PMID- 14527441 TI - Functional organization of adult motor cortex is dependent upon continued protein synthesis. AB - The functional organization of adult cerebral cortex is characterized by the presence of highly ordered sensory and motor maps. Despite their archetypical organization, the maps maintain the capacity to rapidly reorganize, suggesting that the neural circuitry underlying cortical representations is inherently plastic. Here we show that the circuitry supporting motor maps is dependent upon continued protein synthesis. Injections of two different protein synthesis inhibitors into adult rat forelimb motor cortex caused an immediate and enduring loss of movement representations. The disappearance of the motor map was accompanied by a significant reduction in synapse number, synapse size, and cortical field potentials and caused skilled forelimb movement impairments. Further, motor skill training led to a reappearance of movement representations. We propose that the circuitry of adult motor cortex is perpetually labile and requires continued protein synthesis in order to maintain its functional organization. PMID- 14527445 TI - The influence of chain length and electrolyte on the adsorption kinetics of cationic surfactants at the silica-aqueous solution interface. AB - The equilibrium and kinetic aspects of the adsorption of alkyltrimethylammonium surfactants at the silica-aqueous solution interface have been investigated using optical reflectometry. The effect of added electrolyte, the length of the hydrocarbon chain, and of the counter- and co-ions has been elucidated. Increasing the length of the surfactant hydrocarbon chain results in the adsorption isotherm being displaced to lower concentrations. The adsorption kinetics indicate that above the cmc micelles are adsorbing directly to the surface and that as the chain length increases the hydrophobicity of the surfactant has a greater influence on the adsoption kinetics. While the addition of 10 mM KBr increases the CTAB maximal surface excess, there is no corresponding increase for the addition of 10 mM KCl to the CTAC system. This is attributed to the decreased binding efficiency of the chloride ion relative to the bromide ion. Variations in the co-ion species (Li, Na, K) have little effect on the adsorption rate and surface excess of CTAC up to a bulk electrolyte concentration of 10 mM. However, the rate of adsorption is increased in the presence of electrolyte. Slow secondary adsorption is seen over a range of concentrations for CTAC in the absence of electrolyte and importantly in the presence of LiCl; the origin of this slow adsorption is attributed to a structural barrier to adsorption. PMID- 14527446 TI - Kinetics of dodecanoic acid adsorption from caustic solution by activated carbon. AB - This study examines the influences of adsorbent porosity and surface chemistry and of carbon dosage on dodecanoic acid adsorption kinetics from aqueous and 2 M NaOH solutions as batch adsorption processes. Both adsorbents are steam-activated carbons prepared from either coconut or coal precursors. Prior to use the adsorbents were washed in deionized water or 2 M NaOH. Mass transfer coefficients and effective overall diffusion coefficients indicate a minor contribution from adsorbent porosity. In contrast, high surface oxygen content impedes transport to and into the adsorbent structure. Carbon dosage shows a proportional increase in transport coefficients with increasing mass; these coefficients are constant when normalized per unit mass. Neither water nor NaOH treatment of the adsorbents has a significant influence on dodecanoic acid adsorption kinetics. Molecular and Knudsen diffusion coefficients are defined to demonstrate that the overall effective diffusion coefficient values and the diffusion process are controlled by surface diffusion. PMID- 14527447 TI - Nonionic organic solute sorption onto two organobentonites as a function of organic-carbon content. AB - Sorption of three nonionic organic solutes (benzene, trichloroethene, and 1,2 dichlorobenzene) to hexadecyltrimethylammonium bentonite (HDTMA bentonite) and benzyltriethylammonium bentonite (BTEA bentonite) was measured as a function of total organic-carbon content at quaternary ammonium cation loadings ranging from 30 to 100% of the clay's cation-exchange capacity. Sorption of all three solutes to HDTMA bentonite was linear and sorption of all three solutes by the HDTMA bentonite increased as the organic-carbon content of the clay increased. 1,2 Dichlorobenzene sorbed most strongly to HDTMA bentonite, followed by benzene and TCE. The stronger sorption of benzene to HDTMA bentonite compared to TCE was unexpected based on a partition mechanism of sorption and consideration of solute solubility. LogK(oc) values for all three solutes increased with organic-carbon content. This suggests that the increased organic-carbon content alone may not explain the observed increase in sorption capacity. Sorption of the three solutes to BTEA bentonite was nonlinear and solute sorption increased with decreasing organic-carbon content, with a peak in the magnitude of solute sorption occurring at an organic-carbon content corresponding to 50% of CEC. Below 50% of CEC, sorption of all three solutes to BTEA bentonite decreased with decreasing organic carbon content. Surface area measurements indicate that the surface area of both organobentonites generally decreased with increasing organic-carbon content. Since nonionic organic solute sorption to BTEA bentonite occurs by adsorption, the reduced sorption is likely caused by the reduction in surface area corresponding to increased organic-cation loading. PMID- 14527448 TI - Competitive adsorption of Ca2+ and Zn(II) ions at monodispersed SiO2/electrolyte solution interface. AB - A study of competitive adsorption of Ca(2+) and Zn(II) ions at the monodispersed SiO(2)/electrolyte solution interface is presented. Influence of ionic strength, pH, and presence of other ions on adsorption of Ca(2+) and Zn(II) in the mentioned system are investigated. zeta potential, surface charge density, adsorption density, pH(50%), and DeltapH(10-90%) parameters for different concentrations of carrying electrolyte and adsorbed ions are also presented. A high concentration of zinc ions shifts the adsorption edge of Ca(2+) ions adsorbed from solutions with a low initial concentration at the SiO(2)/NaClO(4) solution interface to the higher pH values. This effect disappears with a concentration increase of calcium ions. The presence of Ca(2+) ions in the system slightly affects the adsorption of zinc ions on SiO(2), shifting the adsorption edge toward lower pH values and thereby increasing the adsorption slope. PMID- 14527449 TI - Sorption of Pu(VI) onto TiO2. AB - The sorption of Pu(VI) onto TiO(2) was studied as a function of pH (2-10) and Pu concentration (10(-8)-10(-4) M) under an N(2) atmosphere, in 0.016 and 0.1 M NaClO(4). A batch-wise method was used, in which pH was measured in separate experimental containers after removal of a sample to determine the amount of Pu that had been sorbed. As Pu is radioactive, it was used as a tracer and measured by liquid scintillation counting. No ionic strength dependence was discerned, which was taken as an indication of inner sphere complex formation. In the interval of pH 2-7 the system could be described by the formation of two positively charged surface complexes using a 1-pK Stern model. Sorption of the plutonyl ion (PuO(2)(2+)) and the first hydrolysis species (PuO(2)(OH)(+)) was estimated using FITEQL to logK(1)=6.9 and logK(2)=1.4, respectively. PMID- 14527450 TI - Properties of amphoteric polyurethane waterborne dispersions. II. Macromolecular self-assembly behavior. AB - Amphoteric polyurethane (APU) samples used in this paper were composed of hydrophobic soft segments and pendent -COOH and -CH(2)N(CH(3))(2) groups on the hard segments, which present the properties of both amphoteric polyelectrolytes and amphiphilic block copolymers. APU macromolecules can self-assemble into micelles in acidic and basic aqueous media by hydrophobic/hydrophilic interaction. The self-assembly behavior of APU in acidic and basic media was studied by transmission electron microscopy and light scattering methods. The spherical and hollow micelles of APU were observed respectively in acidic and basic aqueous media. The results indicate that the size and size distribution of APU self-assembly micelles largely depend on the ratio of -COOH to CH(2)N(CH(3))(2) groups, density of ionizable groups, concentration of APU, and types of acid and base in the media. PMID- 14527451 TI - Microencapsulation of oil with poly(styrene-N,N-dimethylaminoethyl methacrylate) by SPG emulsification technique: effects of conversion and composition of oil phase. AB - Microcapsules with narrow size distribution, in which hexadecane (HD) was used as a oily core and poly(styrene-co-N,N-dimethylaminoethyl methacrylate) [P(St DMAEMA)] as a wall, were prepared by a Shirasu porous glass (SPG) emulsification technique and a subsequent suspension polymerization process. That is, a mixture of St monomer, DMAEMA monomer, HD, and N,N'-azobis(2,4-dimethylvaleronitrile) initiator was permeated through the uniform pores of an SPG membrane into a continuous phase containing a poly(vinylpyrrolidone), sodium lauryl sulfate, and sodium nitrite water-soluble inhibitor by applying a pressure, to form uniform droplets. Then, the droplets were polymerized at 70 degrees C. It was found that HD was encapsulated completely only when conversion was quite high, irrespective of whether a DMAEMA hydrophilic monomer was incorporated into the polymer. As the amount of HD increased, HD was more easily encapsulated by the polymer. In order to clarify the reason for such unique behavior, a simulation was carried out, taking the St monomer partition in the HD phase and PSt wall phase into the consideration. It was found that the main reason HD could not be encapsulated completely by PSt when conversion was lower was that the interfacial tension of the HD phase with water and the PSt phase with water got closer. As a result, both HD and PSt can come in contact with the water phase. PMID- 14527452 TI - Thermally responsive polymeric micellar nanoparticles self-assembled from cholesteryl end-capped random poly(N-isopropylacrylamide-co-N,N dimethylacrylamide): synthesis, temperature-sensitivity, and morphologies. AB - Cholesteryl end-capped thermally responsive amphiphilic polymers with two different hydrophobic/hydrophilic chain-length ratios were synthesized from the hydroxyl-terminated random poly(N-isopropylacrylamide-co-N,N-dimethylacrylamide) and cholesteryl chloroformate. The hydroxyl-terminated precursor polymers with narrow molecular weight distributions were synthesized by free-radical polymerization using 2-hydroxyethanethiol as a chain-transfer agent. The aqueous solutions of the cholesteryl end-capped copolymers exhibited reversible phase transitions at temperatures slightly above human body temperature, with the lower critical solution temperature values being 37.7 and 38.2 degrees C, respectively. The critical micelle concentration values of the two cholesteryl end-capped polymers were 9 and 25 mg/L, respectively. Polymeric micellar nanoparticles were prepared from the amphiphilic polymers using a dialysis method as well as a direct dissolution method. Transmission electron microscope studies showed that the micellar nanoparticles existed in different morphologies, including spherical, star-like, and cuboid shapes. Pyrene as a model hydrophobic compound could be readily encapsulated in these polymeric nanoparticles, at loading levels of 1.0 and 0.8 mg/g for the two cholesteryl end-capped polymers, respectively. The temperature sensitivity and unusual morphology of these novel polymeric nanoparticles would make an interesting drug delivery system. PMID- 14527453 TI - Steric interactions between finite objects and end-grafted polymer chains: a two dimensional mean-field analysis. AB - We examine the usefulness of a two-dimensional self-consistent mean-field theory for predicting polymer-induced forces between a finite object and a surface. The predictions are compared with scaling results for the compression of a single, end-grafted chain by a disk and for the compression of a brush with an athermal wall. For the former, the mean-field predictions agree with scaling theory and, in addition, provide the necessary prefactor in the scaling expressions for the Helmholtz potential and force of compression. For the brush, the mean-field results agree with the Alexander-de Gennes scaling result at moderate compressions and also provide an analytical expression for the force without any unknown parameters. We also use the mean-field theory to examine the interaction of a model bacterium with a planar substrate and show that steric interactions due to a single polymer chain alone can dominate van der Waals attraction under typical practical conditions and thus prevent bacterial adhesion to the surface. PMID- 14527454 TI - Calcium modulates the mechanical properties of anionic phospholipid membranes. AB - Using micropipette aspiration and fluorescence techniques, we have studied the material properties of charged lipid vesicles in calcium solutions. Vesicles were composed of phosphatidylglycerol (PG)/phosphatidylcholine (PC) or phosphatidic acid (PA)/PC mixtures. For the case of PG/PC membranes, we measure no effect of anionic lipid fraction on elasticity but a monotonic decrease up to 20% for tension required to induce membrane failure. Both of these observations are rationalized by a model we have developed to describe membrane electrostatic interactions in a two-component salt solution and the resulting changes in membrane properties. Critical tensions measured for PA/PC membranes, on the other hand, did not depend on anionic lipid fraction and were uniformly approximately 35% lower than PG/PC vesicles. This is likely due to a lateral phase separation in the membrane. By combining mechanical properties with fluorescence observations we propose that the PA-rich phase separates into small unconnected domains. PMID- 14527455 TI - Synthesis of nanosized BaSO4 and CaCO3 particles with a membrane reactor: effects of additives on particles. AB - Nanosized BaSO(4) particles, about 15 nm in size, were synthesized successfully by a membrane reactor at the aids of additives, in which Na(2)SO(4) solutions were added into BaCl(2) solutions gradually through the micropores of ultrafiltration membranes to control the saturation ratio, subsequently the nucleation and growth rates. The effects of additives species, additives concentrations, and membranes molecular weight cut-off (MWCO) on the particle morphology, along with the formation processes of particles, were investigated. CaCO(3) nanoparticles of 30-60 nm in size were also prepared by the reactor. The results revealed that the addition of methyl alcohol, ethanol etc. favor the synthesis of nanoparticles with small size. The particles size decreases with the increase in ethanol concentrations. With the increase in membrane MWCO, the products tend from nanoparticles towards aggregates. PMID- 14527456 TI - Flocculation of hematite particles by a comparatively large rigid polysaccharide: schizophyllan. AB - We studied the flocculation kinetics and structure of hematite aggregates induced by a large rigid extracellular polysaccharide, schizophyllan. Transmission electron microscopy (TEM), atomic force microscopy (AFM), photon correlation spectroscopy (PCS), and static light scattering (SLS) were used to characterize hematite particles, schizophyllan chains, and their flocs, to follow the time evolution of floc sizes, and to determine floc fractal dimensions. A maximum flocculation rate was found at a certain schizophyllan/hematite ratio. The maximum rate was considerably smaller than the rate of diffusion-limited aggregation (DLA) of hematite particles induced by simple electrolytes. To interpret the experimental results and to reveal various factors affecting the optimal dosage, Monte Carlo simulations were performed on the flocculation of small colloidal particles by relatively long, monodisperse linear polymers. The existence of the maximum flocculation rate was confirmed by computer simulation. However, a higher optimal dosage of schizophyllan was obtained in the experiments. The difference in the optimal dosage can be attributed mostly to the higher adsorption affinity of the hematite on schizophyllan aggregates present in the initial solution and the presence of a large fraction of free polymer chains which do not participate in the flocculation process. Both experiments and computer simulations demonstrated the fractal nature of the schizophyllan hematite flocs. The fractal dimensions of the flocs at the optimal dosage were determined. A higher fractal dimension was obtained from experiments than from computer simulations, suggesting a reconstruction of the floc structure. Finally, a two-stage flocculation mechanism for hematite particles in the presence of a relatively long schizophyllan polymer was proposed. In the first flocculation stage, the hematite particles are preferentially adsorbed onto the schizophyllan aggregates in solution. The second stage consists of the association of these reactive entities with each other and also with naked chains to form fractal flocs by a bridging mechanism, where the hematite particles play the role of ligands. PMID- 14527457 TI - Synchrotron SAXS/WAXD and rheological studies of clay suspensions in silicone fluid. AB - Suspensions of two commercial smectite clays, montmorillonite KSF and montmorillonite K10, in a low-viscosity silicone oil (Dow Corning 245 Fluid) were studied by simultaneous synchrotron small-angle X-ray scattering (SAXS)/wide angle X-ray diffraction (WAXD) techniques and rheological measurements. In the 0.5% (w/v) KSF clay suspension and two K10 clay suspensions (0.5% and 1.0%), WAXD profiles below 2theta=10.0 degrees did not display any characteristic reflection peaks associated with the chosen montmorillonite clays, while corresponding SAXS profiles exhibited distinct scattering maxima, indicating that both clays were delaminated by the silicone oil. In spite of the large increase in viscosity, the clay suspensions exhibited no gel characteristics. Dynamic rheological experiments indicated that the clay/silicone oil suspensions exhibited the behavior of viscoelasticity, which could be influenced by the type and the concentration of the clay. For the K10 clay suspensions, the frequency-dependent loss modulus (G") was greater in magnitude than the storage modulus (G') in the concentration range from 0.5 to 12.0%. The increase in the clay concentration shifted the crossover point between G' and G" into the accessible frequency range, indicating that the system became more elastic. In contrast, the KSF clay suspension exhibited lower G' and G" values, indicating a weaker viscoelastic response. The larger viscoelasticity response in the K10 clay suspension may be due to the acid treatment generating a higher concentration of silanol groups on the clay surface. PMID- 14527458 TI - Size, volume fraction, and nucleation of Stober silica nanoparticles. AB - 29Si NMR, small-angle X-ray scattering (SAXS), and dynamic light scattering (DLS) are used to monitor the synthesis of silica nanoparticles from the base-catalyzed hydrolysis of TEOS in methanol and ethanol. The reactions are conducted at a [TEOS] =0.5 M, low concentrations of ammonia ([NH(3)] =0.01-0.1 M), and [H(2)O] =1.1-4.4 M to resolve the initial size of the first nuclei and to follow their structural evolution. It is found that after an induction period where there is a buildup of singly hydrolyzed monomer, the first nuclei are fractal and open in structure. Interestingly, the nuclei are twice as large in ethanol (R(g) approximately 8 nm) as those in methanol (R(g) approximately 4 nm). The data suggest that the difference in primary particle size is possibly caused by a higher supersaturation ratio of the singly hydrolyzed monomer in methanol than in ethanol if it is assumed that the surface energy of the first nuclei is the same in methanol and ethanol. The particle number concentration and the volume fraction of the silica particles are calculated independently from the SAXS, DLS, and 29Si NMR results. Finally, the rate of nucleation is obtained from the particle number concentrations. PMID- 14527459 TI - Supersaturation control in aragonite synthesis using sparingly soluble calcium sulfate as reactants. AB - Sparingly soluble calcium salts were studied as reactants in the synthesis of needle-like precipitated calcium carbonate (PCC). The morphology and aspect ratio of the PCC particles were characterized with SEM. Polymorphs and crystal size were characterized using X-ray diffraction. The counterions of the sparingly soluble salts influenced the growth kinetics of PCC as well as the polymorphism and morphology of product particles. Either chrysanthemum-like or needle-like aragonite can be synthesized from calcium sulfate and sodium carbonate depending on the supersaturation and synthesis conditions. Low concentration and slow addition rate of sodium carbonate solution were favorable to the formation of aragonite. Addition of sodium sulfate to the reaction system (calcium chloride and sodium carbonate) promoted the formation of aragonite and decreased the crystal size of aragonite due to the decrease of supersaturation and adsorption of sulfate ion. Too much added sodium sulfate, however, did not further increase the aragonite fraction. An optimal temperature for the formation of aragonite was found to be ca. 60 degrees C. Slow dissolution kinetics of sparingly dissoluble salt also is very important for controlling PCC polymorphism and morphology. PMID- 14527460 TI - Magnitude and direction of thermal diffusion of colloidal particles measured by thermal field-flow fractionation. AB - In this paper we provide experimental evidence showing that various types of submicrometer-sized particles (latexes, inorganic, and metallic), suspended in either aqueous or nonaqueous carrier liquids to which a temperature gradient dT/dx is applied, experience a force in the direction opposite to that of dT/dx. This behavior is similar to that of small particles such as soot, aerosols, and small bubbles suspended in stagnant gases across which temperature gradients are applied, a phenomenon known as "thermophoresis in gases." We report the use of a thermal field-flow fractionation (ThFFF) apparatus in two different configurations to establish the direction of particle motion subject to a temperature gradient. The first approach employed the conventional horizontal ThFFF channel orientation. In this case, small electrical potentials were applied across the narrow channel thickness either to augment or to act in opposition to the applied thermal gradient, depending on whether the accumulation wall was maintained at a positive or negative potential relative to the depletion wall. Thus, by observing the changes in the retention behavior of surface-charged latices or silica particles with changes in potential difference across the channel thickness, we were able to ascertain the direction of migration of the particles in the thermal gradient. The second approach involved the use of a ThFFF column oriented vertically in an implementation of a technique known as thermogravitational FFF. In this approach, the convective flow along the channel length (due to density gradients associated with the temperature gradient) couples with the thermal diffusion effect across the channel thickness to result in a combined particle retention mechanism. A retarded upward migration rate is indicative of accumulation of particles at the cold wall, while enhanced upward migration would indicate a hot-wall accumulation. From the results of our investigations, we conclude that submicrometer-sized particles suspended in either aqueous or nonaqueous carrier liquids and subjected to a temperature gradient migrate from the hot wall toward the cold wall of a ThFFF channel. PMID- 14527461 TI - Size dependence of second-order optical nonlinearity of CdS nanoparticles studied by hyper-Rayleigh scattering. AB - The second-order optical nonlinearity of CdS nanoparticles with different mean diameters of 28.0, 30.0, 31.5, 50.0, and 91.0 A was studied by the incoherent hyper-Rayleigh scattering technique. Results show that the first-order hyperpolarizability beta value per CdS particle decreases with decreasing size from 91.0 to 31.5 A; however, as CdS particle size further decreases, this trend is reversed and the beta value increases. Substantially, the normalized value of the first-order hyperpolarizability per CdS formula unit, beta(0), exhibits systematic enhancement with decreasing size. This is interpreted in terms of a so called surface contribution mechanism. The two aspects of quantum size effects, the size dependence of optical band-gap and oscillator strength, cannot be adopted to explain the size dependence of the second-order optical nonlinearity of CdS nanoparticles. PMID- 14527462 TI - Studies on thermal dehydration of hydrated chromic oxide. AB - Hydrated chromic oxide was prepared by the reduction of potassium dichromate in an autoclave using sucrose. The thermal dehydration of the parent gel in different atmospheres was studied comparatively using the techniques of thermal gravimetry, differential thermal analysis, X-ray diffraction analysis, FT-IR spectroscopy, and gas adsorption. The results of thermogravimetric and differential thermal analysis show that the temperature at which the glow phenomenon occurs is dependent on the gaseous atmosphere in the heat-treatment process and increases in the following order: air, nitrogen, and carbon dioxide. When heated in CO(2), the parent gel is transformed into crystalline alpha Cr(2)O(3) going through the phase of amorphous Cr(2)O(3). In air, however, it is decomposed directly into crystalline alpha-Cr(2)O(3) without going through the amorphous Cr(2)O(3) phase. It is also found that microcrystallites of alpha Cr(2)O(3) have been formed before the glow phenomenon occurs at 390 degrees C in air and at 590 degrees C in CO(2), respectively. The mechanism for thermal dehydration in air and CO(2) is discussed. PMID- 14527463 TI - Structural characteristics of modified activated carbons and adsorption of explosives. AB - Several series of activated carbons prepared by catalytic and noncatalytic gasification and subsequent deposition of pyrocarbon by pyrolysis of methylene chloride or n-amyl alcohol were studied by FTIR, chromatography, and adsorption methods using nitrogen and probe organics (explosives). The relationships between the textural characteristics of carbon samples and the recovery rates (eta) of explosives on solid-phase extraction (SPE) using different solvents for their elution after adsorption were analyzed using experimental and quantum chemical calculation results. The eta values for nitrate esters, cyclic nitroamines, and nitroaromatics only partially correlate with different adsorbent parameters (characterizing microporosity, mesoporosity, pore size distributions, etc.), polarity of eluting solvents, or characteristics of probe molecules, since there are many factors strongly affecting the recovery rates. Some of the synthesized carbons provide higher eta values than those for such commercial adsorbents as Hypercarb and Envicarb. PMID- 14527464 TI - Electrochemical studies on surfactant-modified cellulose acetate membrane. AB - The possibility of electrochemical modification of cellulose acetate membrane upon immobilization of the anionic surfactant (SDS) has been explored on the basis of membrane potential studies. Surface tension measurements with and without cellulose acetate membrane were carried out to ascertain the extent of immobilization of the surfactant. Cellulose acetate membrane practically does not exhibit any ion selectivity. However, modified membrane exhibits cation selectivity which varies with concentration of the surfactant till its critical micelle concentration is reached. An attempt has also been made to demonstrate correspondence between the immobilized surfactant and the permselectivity of the membrane. PMID- 14527465 TI - Dynamic viscoelastic properties of spread monostearin monolayer in the presence of glycine. AB - The monostearin monolayer at the air-aqueous interface is more expanded in presence of glycine and at higher temperature from both the surface pressure-area per molecule (pi-A) isotherms and static elasticity-surface pressure (E(s)-pi) curves. The dilational viscoelastic properties of monostearin monolayer spread on the subphase of glycine solution have been determined by the dynamic oscillation method and discussed as a function of surface pressure, temperature, and frequency. At the frequency of 50 mHz, the monostearin monolayer on pure water shows negative dilational viscosity and is viscoelastic at some surface pressures, while the monostearin monolayer in the presence of glycine is nearly elastic over a wide range of surface pressure, especially at 25 degrees C. Both positive and negative loss angle tangent can be deduced as a function of surface pressure. The negative dilational viscosity can be attributed to the phase transitions induced by the propagation of the surface waves during the dynamic oscillation. It can be convinced that the interactions between monostearin and glycine play an important role in the formation and rheological behavior of the monolayer. On the other hand, temperature has effect on the dilational elasticity and the dilational viscosity of the monostearin monolayer in different extents. Furthermore, at the surface pressure of 20 mN/m, the monostearin monolayer on the glycine solution at 18 degrees C is essentially elastic at lower frequency (<100 MHz) and shows viscoelastic behavior at higher frequency. These phenomena should be associated with the complicated monolayer structure and structural reorganization due to the interactions between monostearin and glycine in presence of glycine. PMID- 14527466 TI - Effect of solid walls on spontaneous wave formation at water/oil interfaces. AB - The regulation of spontaneous waves at water/oil interfaces was investigated, focusing on effects of materials and sizes of containers. Trimethylstearylammonium chloride was dissolved in an aqueous phase. Nitrobenzene with potassium iodide and iodine was used as an organic phase. Rotation of interfacial waves with almost triangular shape was observed only in containers made of glass. The nature of interfacial waves is sensitive to container size. There was no interfacial wave in PFA (Teflon) containers. However, when a glass plate was soaked vertically to the interface, oscillation of contact angles of water/oil interfaces to glass plates was observed. The oscillation generated wave propagation along the plate. Dynamic interfacial tension was measured by Wilhelmy method and the pendant drop technique. Results with the Wilhelmy method in small glass containers exhibited spontaneous oscillation. However, oscillations in dynamic interfacial tension were not observed for other cases, i.e., the Wilhelmy method for large glass containers, for PFA containers, and for the pendant drop technique. It was concluded that all nonlinear behavior such as wave generation and apparent tension oscillation could be attributed to the effect of the sidewalls of container on the adsorption/desorption kinetics of the surfactant. We propose a possible scenario which can explain all of the qualitative features of the present experimental findings. PMID- 14527467 TI - Enthalpy of hydrophobic hydration separately estimated for each -CH2-, -CF2-, and -OCH2CH2- unit component of amphiphiles and of related compounds. AB - The enthalpy of hydrophobic hydration, DeltaH(HH), was separately estimated for each apolar segment of amphiphiles and of related compounds. The specimens analyzed in this study were the homologs and analogs of sodium alkyl sulfate, alkyltrimethylammonium bromide, ammonium perfluoroalkylcarboxylate, and polyethylene glycols. The DeltaH(HH) was calculated from the adiabatic compressibility beta of aqueous solution systematically measured for each homologs at varying concentrations and temperatures. Those data reported in the previous paper were utilized. The DeltaH(HH) estimated for each -CH(2)-, -CF(2)-, and -OCH(2)CH(2)- segments in both hydrophobic and hydrophilic parts of the amphiphiles was found to change depending on its locations in the molecule. The DeltaH(HH) for the first -CH(2)- unit which is adjacent to the head sulfate group of H(CH(2))(n)SO(4)Na was found to be negligibly small; however, it increases gradually as the position of the unit moves apart from the anionic head group and attains as large as -26.5 kcal/mol for the eighth -CH(2)-. The change of DeltaH(HH) of -CF(2)- unit in F(CF(2))(n)COONH(4) shows the similar tendency to H(CH(2))(n)SO(4)Na. The value of DeltaH(HH) is as small as -8 kcal/mol for the first -CF(2)-, however, it increases to -16 kcal/mol for the fifth -CF(2)-, which is 1.5 times larger than that of the fifth -CH(2)- of alkyl sulfate. Contrary to this, all the DeltaH(HH) of the methylenes of cationic surfactant ranging from the second to the eighth -CH(2)- units of H(CH(2))(n)N(CH(3))(3)Br take a similar value around -8 kcal/mol of -CH(2)- irrespective of their positions. The first CH(2)- unit which is equivalent to that of quaternary ammonium [HCH(2)](4)NBr shows slightly higher value of -9.0 kcal/mol of the first -CH(2)- than those for the second to eighth -CH(2)-. This seems to suggest that the cationic core nitrogen has an enhancing effect on hydrophobic hydration. The hydration enthalpy DeltaH(OEH) of each oxyethylene (OE) unit of polyethylene glycol was also estimated for H(OCH(2)CH(2))(p)OH (p=1-4) homologues to see the roles of ether oxygen and hydroxyl groups in hydrophobic hydration. The value of DeltaH(OEH) showed its maximum -13.1 kcal/mol for the OE unit located at the middle of three. The other two units at both ends were found to take the value of -9.9 kcal/mol, smaller than that at the center. This could suggest that the hydrophobic hydration of the ethylene segment is largely enhanced by the neighboring ether oxygens. PMID- 14527468 TI - Feasibility of surfactant-free supported emulsion liquid membrane extraction. AB - Supported emulsion liquid membrane (SELM) is an effective means of conducting liquid-liquid extraction. SELM extraction is particularly attractive for separation tasks in the microgravity environment where density difference between the solvent and the internal phase of the emulsion is inconsequential and a stable dispersion can be maintained without surfactant. In this research, dispersed two-phase flow in SELM extraction is modeled using the Lagrangian method. The results show that the SELM extraction process in the microgravity environment can be simulated on earth by matching the density of the solvent and the stripping phase. The feasibility of surfactant-free SELM (SFSELM) extraction is assessed by studying the coalescence behavior of the internal phase in the absence of the surfactant. Although the contacting area between the solvent and the internal phase in SFSELM extraction is significantly less than the area provided by regular emulsion due to drop coalescence, it is comparable to the area provided by a typical hollow-fiber membrane. Thus, the stripping process is highly unlikely to become the rate-limiting step in SFSELM extraction. SFSELM remains an effective way of achieving simultaneous extraction and stripping and is able to eliminate the equilibrium limitation in the typical solvent extraction processes. The SFSELM design is similar to the supported liquid membrane design in some aspects. PMID- 14527469 TI - Stable water-oil emulsions from the new insoluble surfactant sodium 5-(1 dodecylaminocarbonyl) picolinate. AB - The new compound sodium 5-(1-dodecylaminocarbonyl) picolinate (NaDPA) has been prepared and found to have the unusual surfactant property of forming stable water-oil (w/o) emulsions in the presence of water and a variety of organic liquids. A study of droplet size (diameters=0.1-1.0 mm) as a function of the amount of surfactant used shows that a near constant coverage of surfactant molecules exists at the water-oil interface and that the interface is made up of multiple layers. The structure of NaDPA, with its multiple hydrogen-bonding possibilities, probably contributes to the observed stability of the emulsions. The emulsions are made up of droplets that are stable for months and can be filled with dyes, buffers, and solutions of high ionic strength. PMID- 14527470 TI - Effects of a bolaamphiphile on the structure of phosphatidylcholine liposomes. AB - This paper describes the morphological characterization of aqueous dispersions of PC amphiphiles and bolaamphiphile AEC by microscopy, the liposomal membrane fluidity, and the zeta potential. Results indicate that the bolaamphiphile AEC can be included within conventional egg-PC liposome bilayers. This behavior could be due to their preference for the stretched conformation within the PC membranes. PMID- 14527471 TI - A dynamic loading method for controlling on-chip microfluidic sample injection. AB - A new technique for controlling discrete sample injection in straight-cross microfluidic chips is presented here. This technique involves a three-part process with a dynamic loading step in between the steady-state loading step and the dispensing step. During the intermediate step, sample is pumped into the intersection and into the three connecting channels. The key features of this technique are the ability to dynamically control the sample size and the ability to inject well-defined samples at the original sample concentration. Injections of these samples with lengths varying from 2 channel widths (100 microm) to 20 channel widths (millimeter-sized) are demonstrated. The sample concentration profiles obtained are compared with those of focused and less-focused pinched valve injections. In applications such as high-speed capillary zone electrophoresis, this technique can provide an increase in signal with a small increase in sample length. This technique is especially applicable to many large sample applications in which the offset twin-T microchip has been previously employed. PMID- 14527472 TI - Cadmium sulfide nanocrystals via two-step hydrothermal process in microemulsions: synthesis and characterization. AB - CdS nanocrystals with an average diameter of 16 nm were synthesized in the CTAB/n C(5)H(11)OH/n-C(6)H(14)/water quaternary microemulsions by a two-step hydrothermal process at 90 and 130 degrees C. The reaction of carbamide and carbon disulfide was employed as the sulfur source for the preparation of CdS nanocrystals. The resulting crystals were characterized with powder X-ray diffraction, transmission electron microscopy, UV-vis absorption spectroscopy, and photoluminescence spectroscopy. A unique core/shell structure of CdS nanocrystals was suggested for the explanation of the interesting phenomenon. PMID- 14527473 TI - Viscosity of concentrated colloidal suspensions: comparison of bidisperse models. AB - In this paper, recent advances in the study of rheological behavior of concentrated bimodal suspensions are briefly reviewed. The predictive models are divided into two categories, namely, the effective volume fraction (or hard sphere scaling) approach and the separation of contributions approach. Predictions of both approaches are compared with experimental data of electrostatically and sterically stabilized suspensions. It is shown that the predictions of both hard sphere scaling and the scaling method of Zaman and Moudgil (J. Colloid Interface Sci. 212 (1999) 167) to separate the contributions of fine and coarse particles are in good agreement with the experimentally observed results. The approach by Dames, Morrison, Wilenbacher (Rheol. Acta 40 (2001) 434) to separate the hard-sphere and non-hard-sphere contributions is investigated using the aqueous silica and polystyrene suspensions respectively. A good agreement is shown for aqueous silica suspensions. However, significant differences between the predictions and experimental data are found for the sterically stabilized polystyrene suspensions, suggesting a more generalized expression is needed. As an attempt to classify the models on the viscosity of colloidal suspensions, the present study will provide guidelines for interpretation of experimental results and for the development of more comprehensive predictive methodologies for polydispersed colloidal dispersions. PMID- 14527474 TI - Charge variance of ionic micelles: a significant parameter to account for and to predict changes in degrees of micelle ionization and in aggregation numbers. AB - In the context of the thermodynamics of self-assembling systems, which predicts that both the number-average number of aggregation, N(n), of the ionic surfactants and the number-average degree of ionic dissociation of the micelles, alpha(n), depend on the concentrations of micellized surfactant and counterion in the bulk, it is shown that a key parameter is the ratio between sigma(Q), the standard deviation of the micelle charge distribution of the micelles, and sigma(N), that of the aggregation numbers. In the case of hexadecyltrimethylammonium acetate-acetic acid (1:1) solutions, this ratio is calculated as ca. 0.1 from conductivity and aggregation number data. PMID- 14527475 TI - Developing the solution analogue of the Toth adsorption isotherm equation. AB - The well-known Toth adsorption isotherm equation developed formerly for adsorption of vapors is converted into its solution analogue. It is shown that this equation can be successfully applied to the description of adsorption data of organics on activated carbons. PMID- 14527476 TI - Formation of tubules and giant vesicles from large multilamellar vesicles. AB - This study reports an observation of submicrometer multilamellar vesicles (MLVs) prepared by simply freeze-thawing a phospholipid dispersion at full hydration that transformed into giant vesicles (GVs) and tubules (TUs) when confined between microscope glass slides. Cover slide cleaning and surface treatment did not hamper the formation of GVs or TUs. However, when small unilamellar vesicles (SUV) were prepared or when MLVs were not confined but rather freely moved between the glass slides or when the phospholipid was in its gel phase, neither GVs nor TUs were observed. Altogether, our results suggested that MLVs would play a role as a lipid reservoir and that the liquid flow between the glass slides induces the peeling of the external bilayers, yielding the formation of tubules and giant unilamellar vesicles. PMID- 14527479 TI - Medical comorbidities in the treatment of epilepsy. AB - The treatment of epilepsy extends far beyond seizure control. Many comorbidities have a significant impact on the medical management and quality of life of patients with epilepsy. In this review, we examine interactions between epilepsy and some common medical conditions. Psychiatric disorders with a high prevalence in epilepsy include mood disorders, anxiety disorders, and psychosis. Depression is common, psychosis occurs both in direct relation to seizures and interictally, and suicide rates are increased. Changes in sexual function and reduced fertility and marriage rates are described, including a discussion of polycystic ovary syndrome, which is increased in women with epilepsy. The effects of other chronic medical comorbid conditions are reviewed, including the effects of antiepileptic medications on bone health and the impact of renal insufficiency on pharmacological therapy of epilepsy. PMID- 14527480 TI - Epilepsy and migraine. AB - Migraine and epilepsy are both chronic disorders characterized by recurrent neurologic attacks variously accompanied by headache as well as by gastrointestinal, autonomic, and psychologic features. Migraine and epilepsy are linked by their symptom profiles, comorbidity, and treatment. The presence of one disorder increases the likelihood that the other is also present. In addition, a number of migraine syndromes can be confused with epilepsy. The diagnosis and treatment of each disorder must take into account the potential presence of the other. Herein, we review the relationship between migraine and epilepsy. We discuss the diagnosis of migraine using the International Headache Society criteria, emphasizing the variants of migraine most frequently mistaken for epilepsy. We summarize the epidemiologic evidence that migraine and epilepsy are associated and discuss specific interrelationships between migraine and epilepsy. PMID- 14527481 TI - Epilepsy and cognition. AB - Patients with epilepsy are more prone to cognitive and behavioral deficits. Epilepsy per se may induce or exacerbate an underlying cognitive impairment, a variety of factors contribute to such deficits, i.e., underlying neuropathology, seizure type, age of onset, psychosocial problems, and treatment side effects. Epilepsy treatment may offset the cognitive and behavioral impairments by stopping or decreasing the seizures, but it may also induce untoward effects on cognition and behavior. The neurocognitive burden of epilepsy may even start through in utero exposure to medications. Epilepsy surgery can also induce certain cognitive deficits, although in most cases this can be minimized. Clinicians should consider cognitive side effect profiles of antiepileptic medications, particularly in extreme age groups. While no effective treatments are available for cognitive and behavioral impairments in epilepsy, comprehensive pretreatment evaluation and meticulous selection of antiepileptic drugs or surgical approach may minimize such untoward effects. PMID- 14527482 TI - Epilepsy and sleep disturbance. AB - Sleep disturbance is common in epilepsy, the nature of sleep disturbances in epilepsy is diverse, and the etiologies are complex. Evidence suggests that having epilepsy and the occurrence of seizures, as well as some AEDs, are associated with significant sleep disruption. The occurrence of seizures can have profound effects on sleep architecture lasting much longer than the postictal period. Persistent daytime drowsiness in patients with epilepsy is not always due to the side effects of some AEDs and may be independently linked with sleep fragmentation. Significant sleep disruption in epilepsy has been associated with impaired quality of life and impaired seizure control. All aspects of sleep medicine are important in the management of epilepsy and are confounded by the occurrence of seizures, the location of seizures, and the beneficial and detrimental effects of AEDs. Sleep should be proactively evaluated, and sleep disturbances should be treated as part of the total care of patients with epilepsy. PMID- 14527483 TI - From the Epilepsy Foundation: restoring the Pediatric Rule. PMID- 14527484 TI - Epilepsy: disease, illness, or disorder? PMID- 14527485 TI - Translating monotherapy trials into clinical practice: a look into the abyss. AB - To be approved for monotherapy by regulatory authorities, new antiepileptic drugs (AEDs) must first be tested in well-controlled studies in refractory patients (conversion to monotherapy trials) or in patients with newly diagnosed epilepsy. However, the applicability of the information obtained in these trials to day-to day clinical practice is limited. Clinical trials in newly diagnosed patients, particularly those allowing dose flexibility, offer more useful information, but a close scrutiny of methodological details is required to avoid misinterpretation of the findings. In many instances, the neurologist has a drug with a label, but lacks critical information on optimal titration rates, optimal target and maintenance dosages, response rates in populations with different epilepsy syndromes, different age ranges and comorbidities, and long-term safety data. Such information becomes available only through general clinical experience, well designed phase IV studies, and postmarketing surveillance. PMID- 14527486 TI - Musicogenic epilepsy and epileptic music: a seizure's song. PMID- 14527487 TI - Schizophrenias and epilepsies: why? when? how? AB - Detailed studies of the association between the epilepsies and the schizophrenias extend over 40 years. Recent studies are abundant and make fertile use of new technologies. However, the pathological changes described in schizophrenias are quite varied. Studies fail to recognize that "epilepsies" and "schizophrenias" have varied in definition over time, and have always been weak categories with which to do science. Now that it is possible to measure deficits in brain structure, it would be better to see what behavioral problems are associated with specific cerebral pathology. It would be wise to be very precise in describing the behaviors and the nature and timing of their emergence rather than using terms such as psychoses. Schizophrenias have neurological, neuropsychological, and behavioral antecedents in childhood. Those associated with later epilepsies show biases, persistent in many studies over a number of years, toward relative excess of females, left temporal lobe structural deficits, and non-right handedness. Schizophrenia should now be a predictable eventuality in certain people with epilepsies, an important factor in medical and surgical treatment. PMID- 14527488 TI - Parental anxiety and quality of life in children with epilepsy. AB - Parental beliefs and attitudes concerning epilepsy may significantly impact adjustment and quality of life for both the child and family. The purpose of the present study was to examine the relationship between parental anxiety and quality of life in pediatric patients with ongoing epilepsy. Subjects were parents (n=200) of children between the ages of 6 and 16 years who had been diagnosed and treated for epilepsy for at least 1 year. Parents were given quality of life and anxiety questionnaires during the child's clinic visit. A stepwise regression analysis suggested that severity of comorbid conditions, parental anxiety, seizure control, and number of medications were significantly associated with quality of life for these children. Parents with increased anxiety whose children had poorly controlled seizures and a comorbid disability were found to have diminished quality of life. PMID- 14527489 TI - Postictal serum levels of antiepileptic drugs for detection of noncompliance. AB - Medication noncompliance (NC) is thought to be a major cause of insufficient seizure control. In an explorative study we investigated whether postictal serum levels (PISLs) of antiepileptic drugs (AEDs) are a reliable indicator of NC. Young adults with epilepsy on a stable AED regimen were asked to contact medical service as soon as possible when a seizure occurred to obtain serum levels of their AEDs. PISLs were compared with the mean value of two routine serum levels of the same medication. PISLs lower than 50% of the individual reference value were regarded as an indicator of NC. PISL samples in 61 seizures of 52 patients treated mainly with carbamazepine, valproic acid, or lamotrigine were evaluated. A drop in serum levels >50% indicating NC was noted in 44.3% of the seizures. Determination of PISLs seems to be a simple and useful method for detecting or ruling out irregular intake patterns as a cause of "breakthrough" seizures. PMID- 14527490 TI - A study of nonepileptic seizures in an Indian population. AB - OBJECTIVE: The goal of this study was to evaluate the background and the clinical profile of nonepileptic seizures (NESs) confirmed by short-term video encephalography (ST-VEEG) recording in an Indian population. METHODS: Seventy-one patients with NESs were enrolled. A complete history was taken and the recorded event was reviewed to define the ictal events. Patients were divided into two groups, Group 1 with a paucity of movements and Group 2 with an excess of movement, and results were analyzed. RESULTS: The mean age was 22.9 (9.6); there were 15 males (21.1%) and 56 females (78.9%). Twenty-four patients (33.8%) were receiving antiepileptic drugs. The majority of the patients (42, 59.1%) were students. All patients were amnesic for the event and were unresponsive during the event. The other characteristics were similar events in 98.6%, hyperventilation in 58 (81.7%). Forty-two patients (59.2%) were in Group 1 and 29 patients (40.8%) were in Group 2. CONCLUSION: NES is a disease of the young and can affect the student or professional. A wrong diagnosis can result in inappropriate treatment. Awareness of this entity is critical to ensuring prompt diagnosis and early intervention. PMID- 14527491 TI - Saline infusion: a diagnostic and therapeutic tool in nonepileptic attacks? AB - OBJECTIVE: Nonepileptic attacks (NEAs) pose diagnostic and therapeutic uncertainties. Prognosis is poor. Activation procedures like saline infusion have been used for diagnostic purposes. METHOD: We reviewed 66 consecutive patients with a probable diagnosis of NEA. During the EEG an attempt was made to induce an attack with verbal suggestion and saline infusion. Patients were followed up with a postal or telephone questionnaire 4.5 years after the EEG. RESULTS: Saline infusion with verbal suggestion induced in 41 of 66 patients a NEA, i.e., a positive test. Thirty-one patients were followed up. Attacks were reduced in 20 patients, of whom 12 were attack-free. More patients were attack-free at follow up after a positive (7 of 16) in comparison to a negative (5 of 15) test. CONCLUSION: A provocative test can be diagnostic in some patients with NEA. An early and firm diagnosis may lead to appropriate treatment and better outcome. PMID- 14527492 TI - Evaluation of physical exercise habits in Brazilian patients with epilepsy. AB - In this study we present data from a survey that aimed to assess the physical activity of a sample of adult outpatients with epilepsy. One hundred adult outpatients of both sexes with epilepsy answered a survey addressing exercise habits. Fifty-eight males and forty-two females participated in this study. The mean age of onset of seizures was 18.6 years and the mean duration of epilepsy was 16.1 years. Sixty patients had controlled or rare seizures, 8 infrequent seizures, 17 frequent seizures, and 11 very frequent seizures. Eighty-six had partial epilepsy and only 3 had abnormal neurological examinations. Of the total, 51 engaged in physical activity, 85 did not believe that sports precipitate seizures, and 15 were forbidden by their physicians to engage in physical activities. Moreover, 14 were cautioned against participation in sports by their relatives and friends. Eight-four patients had never experienced seizures during physical exercise, 36 believed that physical activity has a positive influence on treatment, and only 1 related injuries associated with seizures. Forty-five are afraid of having seizures during exercise because the seizures might attract the attention of others and they would make fools of themselves. Our data show that although most of our patients do not regularly engage in physical activity, they believe that it might improve medical treatment. PMID- 14527493 TI - Stress and epilepsy: a patient perception survey. AB - A questionnaire was administered to patients in the Montefiore Medical Center outpatient epilepsy department to assess perceptions about stress, seizures, and stress reduction. Eighty-nine patients completed the questionnaire. Overall, 64% of patients reported the belief that stress increased the frequency of their seizures. This belief was not significantly associated with gender, age, location of care, epilepsy classification, or seizure control, but was significantly associated with a shorter duration of epilepsy (P=0.04). Thirty-two percent of subjects had tried stress reduction modalities for epilepsy. Of those who had not, 53% were willing to try, and this willingness was significantly associated with a greater number of seizures in the prior 2 months (P=0.006) and the belief that stress was associated with seizures (P=0.04). Both major and minor stressors were reported with equal frequency. The majority of our study population believe that stress and seizures are related, and are willing to try stress reduction techniques for seizure control. We believe that these findings indicate a need for a prospective study to evaluate the role of stress and stress reduction as an additional potential therapeutic modality for epilepsy. PMID- 14527494 TI - The influence of comorbid depression on quality of life for people with epilepsy. AB - The impact of depression on health-related quality of life (HRQOL) for people with epilepsy was evaluated using a postal survey that assessed HRQOL, depression, and seizure severity (N=501). QOLIE-89 scores were significantly reduced by comorbid depression (all P<0.0001) for all types of seizures. People with predominantly major, partial, and minor seizures had significantly poorer HRQOL if they had mild-moderate or major depression (all P<0.0001). People with any type of recent seizure had significantly poorer HRQOL in all domains than people who were seizure-free for 3 months. Differences between groups by depression category were clinically significant (>12 points). Depression (r= 0.72), seizure bothersomeness (r=-0.54), seizure severity (r=-0.37), and days disabled with seizures (r=-0.65) were significantly correlated with poorer HRQOL in all domains than found for nondepressed patients (all subscales P<0.0001). Clinical depression is significantly associated with poorer HRQOL among people with all types of seizures. PMID- 14527495 TI - Attention, memory, and behavioral adjustment in children with frontal lobe epilepsy. AB - To explore whether attention, memory, and behavior would be more affected in children with frontal lobe epilepsy than in children with other types of epilepsy, we compared 16 children with frontal lobe epilepsy (FLE), 8 with temporal lobe epilepsy (TLE), and 8 with generalized absence (GEA) seizures on the Performance Speed (PS) and Freedom of Distraction (FD) indices of the WISC III, the Continuous Performance Test (CPT), the California Verbal Learning Test (CVLT), and Rey-Osterrieth Complex Figure (ROCF). Parents completed Achenbach's Child Behavior Check List. Children with FLE scored significantly lower than the other two groups on the PS and CPT. On the CVLT they made more intrusion errors and were more prone to interference. Furthermore, they had more difficulties copying and recalling the ROCF. Behavior profiles revealed greater attention problems in this group. This may put children with FLE at greater risk of developing school problems than children with TLE and GEA. PMID- 14527496 TI - Aggressive behavior of epilepsy patients in the course of levetiracetam add-on therapy: report of 33 mild to severe cases. AB - Levetiracetam (LEV) was shown to be very efficacious and well tolerated as add-on therapy for refractory epilepsy. Here we report 33 patients with longstanding histories of epilepsy who experienced aggressive episodes during LEV therapy. This corresponds to 3.5% of LEV-treated patients as compared with less than 1% of patients not on LEV. Among these cases, 24 showed only moderate, partly transient irritability, with 10 patients requiring reduction or discontinuation of LEV. More strikingly, 9 patients displayed severe symptoms of aggression with physical violence and, in 2 cases, the need for psychiatric emergency treatment. One patient developed additional psychotic symptoms. We suggest that, specifically in patients with a previous history of aggression, behavioral tolerability of LEV should be carefully monitored. PMID- 14527497 TI - A past psychiatric history may be a risk factor for topiramate-related psychiatric and cognitive adverse events. AB - OBJECTIVES: Topiramate (TPM) is a new antiepileptic drug (AED) that has been found to be associated with a high prevalence of cognitive adverse events (CAEs). The prevalence of psychiatric adverse events (PAEs) has yet to be established. The purpose of this study was to determine the prevalence of PAEs related to TPM when used in polytherapy regimens in a large cohort of adult patients with epilepsy, to identify any association between the occurrences of CAEs and PAEs and to identify predictors of PAEs and CAEs. METHODS: Investigators from 16 epilepsy centers (PADS group) prospectively obtained postmarketing safety and efficacy data on 596 patients aged 16 years and older. All data were recorded on standardized data retrieval forms, completed at the initial visit, while follow up data were obtained every 6 months or at the time of discontinuation. RESULTS: PAEs were identified in 75 (12.6%) patients: 30 (5%) experienced symptoms of depression and 34 (5.7%) of aggressive behavior and irritability, while 9 patients experienced symptoms of psychosis (1.5%). CAEs were reported by 247 (41.5%) patients. There was a significant association between the occurrences of CAEs and PAEs. A past psychiatric history was a predictor of CAEs, while older age and past psychiatric history were predictors of PAEs. CONCLUSIONS: The use of TPM in polytherapy regimens can cause PAEs and CAEs and their occurrence is significantly correlated. Patients with a past psychiatric history may be at a higher risk for experiencing PAEs and CAEs. PMID- 14527498 TI - Treadmill injuries in patients with epilepsy. PMID- 14527499 TI - Pre- and postoperative intracarotid amytal procedure: an assessment of validity. AB - The intracarotid amytal procedure (IAP) was used twice to assess the suitability of three male patients for two successive neurosurgical procedures to relieve intractable temporal lobe epilepsy. First an amygdalohippocampectomy was performed, then further tissue was removed in a temporal lobe resection because their seizures had failed to remit. Repetition of the IAP following amygdalohippocampectomy when there was a known excision allowed inferences to be made regarding its validity in assessing lateralization of language functioning, memory functioning, and lateralization of seizure focus. The procedure was found to be reliable in assessing both language dominance and adequacy of memory functioning of the hemisphere contralateral to the lesion site. The procedure's third function of lesion lateralization was valid for identifying the known neurosurgical lesion. However, it was less successful in corroborating the lateralization of seizure focus before amygdalohippocampectomy. Differences in cognitive outcome between the two neurosurgical procedures are discussed. PMID- 14527500 TI - Tourette's syndrome following temporal lobectomy for seizure control. AB - Gilles de la Tourette's syndrome (TS) is a neurobehavioral disorder characterized by multiple motor and vocal tics, occurring longer than a year and causing marked distress along with social and occupational impairments in level of functioning. It can be accompanied by obsessive-compulsive behavior and attention deficit disorder. This report discusses the case of a young woman with a simple motor tic disorder and intractable seizures who, after right temporal lobectomy for medically intractable epilepsy, developed TS with complex motor and vocal tics, severe obsessive-compulsive disorder, and paranoia. This neurobehavioral complication has not to our knowledge been previously reported after epilepsy surgery. PMID- 14527501 TI - Transient postoperative prosopagnosia. AB - A 23-year-old right-handed woman developed isolated transient prosopagnosia following surgical resection of a right posterior temporal seizure focus. At 18 years of age she had developed secondarily generalized tonic-clonic seizures. Preoperative neuropsychological evaluation was normal, and neurological examination revealed only a left superior quadrant achromatopsia. MRI revealed a circumscribed lesion in the right inferolateral temporo-occipital junction. Following surgery she was agitated for 36 hours, and afterward, when her attention and orientation improved, she was unable to recognize familiar faces. She could, however, recognize familiar voices. Her prosopagnosia resolved over the next 6-7 days. This case demonstrates that isolated prosopagnosia can occur in patients with lesions restricted to the right inferior posterior temporal anterior occipital region. The temporary nature of the prosopagnosia may result from postsurgical tissue injury, including focal cerebral edema, with compensation by ipsilateral or contralateral areas. PMID- 14527503 TI - Laterality and psychopathology circa 1976. PMID- 14527502 TI - Landau-Kleffner syndrome responsive to levetiracetam. AB - A 5-year-old girl with Landau-Kleffner syndrome is discussed. The child began having seizures at age 4 associated with language deterioration despite anticonvulsant therapy. With levetiracetam monotherapy to a dose of 60 mg/kg/day and discontinuation of carbamazepine and valproic acid, her language has improved and seizures are controlled. Levetiracetam should be considered as therapy for Landau-Kleffner syndrome. PMID- 14527504 TI - Lateralized temporal-limbic dysfunction and psychopathology. 1976. PMID- 14527505 TI - Sedation caused by primidone may exacerbate dementia. PMID- 14527506 TI - Fasting and its biblical use related to epilepsy and demonic possession. PMID- 14527507 TI - Neonatal exposure to propylthiouracil induces a shift in lymphoid cell sub populations in the developing postnatal male rat spleen and thymus. AB - Evidence of the connections between the immune system and the thyroid axis is increasingly strong; however, much of the data are focused on immune effects of altered thyroid status in adults or rodents with congenital defects of the pituitary/thyroid axis. The object of the present study was to determine the effects of PTU-induced hypothyroidism on the developing immune system of the rat by focussing on both the spleen and thymus gland. Male Sprague-Dawley rat pups were exposed to PTU through maternal milk by giving the mothers 0.02% PTU in their drinking water starting on the pups' day of birth until day 24 (d24), shortly before weaning on d28. Animals were sampled on days 14, 22, 30, and 91. The mean body weight was decreased in the PTU-treated animals on days 14, 22, and 30. The mean spleen and thymic weights and cellularity were all decreased in the PTU-treated animals on d22 and d30. PTU exposure increased the proportion of NK cells in the spleen on days 14, 22, and 30. The proportion of T-cells was increased on days 22 and 30 with a particular increase in the CD4+ T-cells, resulting in an increase in the ratio of helper T-cells to suppressor/cytotoxic T cells at d22. PTU also decreased the proportion of splenic B-cells at days 14, 22, and 30 which could explain the increased proportion of both NK and T-cells during these sampling periods. PTU treatment decreased the lytic ability of NK cells at d22, but no functional differences were observed at days 14, 30, 91, despite the increased proportion of NK cells in PTU-exposed animals at days 14, 22, and 30. PTU exposure also increased the proportion of CD4+CD8- cells in the thymus on d22 and caused an increase in both the CD4+CD8- and CD4-CD8+ populations on d30. These data suggest that the effects of temporary, PTU-induced hypothyroidism on the cell populations in the spleen partially result from transient changes in thymic T-cell development, including a shift towards increased CD4+CD8- cells. The data also suggest that temporary hypothyroidism early in development decreases B-cell development in a transient fashion. Temporary hypothyroidism induced from birth to the latter stages of the weaning period induced transitory effects on the spleen, thymus, and immune cell sub populations--all of which recovered to normal values when the animals matured. PMID- 14527508 TI - Tolerance induced by anti-CD3 immunotoxin plus 15-deoxyspergualin associates with donor-specific indirect pathway unresponsiveness. AB - Peritransplant treatment with anti-CD3 immunotoxin plus deoxyspergualin induces tolerance to kidney allografts in most rhesus macaque recipients. Tolerant recipients maintain normal function for years without evidence of chronic rejection. Indirect alloantigen presentation is implicated in chronic rejection. Accordingly, we determined if anti-CD3 immunotoxin plus deoxyspergualin induced rejection-free tolerance associates with suppression of anti-donor indirect pathway responses. Tolerant recipients exhibited an early decrease in direct anti donor responses with recovery to baseline levels by 3 years posttransplantation. In contrast, tolerant monkeys were unresponsive to donor antigens presented by the indirect pathway. Recipients that rejected their allografts retained vigorous direct and indirect anti-donor responses. Therefore, following temporary donor specific hyporesponsiveness, direct responses recover in tolerant recipients >1.5 years after transplantation. However, tolerant recipients tested at 1.9-4 years posttransplant are specifically unresponsive to donor antigens presented by the indirect pathway. Thus, the rejection-free state of tolerant recipients may depend on mechanisms regulating indirect pathway responsiveness. PMID- 14527509 TI - Early activation events differentiate the reactivity of two T-cell families to Staphylococcus enterotoxin A. AB - Analysis of early activation events in two SEA responsive T-cell families demonstrated that low doses of SEA induced CD4+Vbeta22 T-cells to down-regulate their TCR and express CD69, considerably earlier than CD4+Vbeta5 T-cells. The rapid down-regulation of Vbeta22 TCR led to its proliferation, whereas even a 10 fold higher dose of toxin induced only a partial down-regulation of Vbeta5 TCR. Stimulation with SEA induced a significantly higher percentage of Vbeta22 T-cells to produce IFN-gamma compared to Vbeta5 T-cells. SEAF47A, a mutant of SEA, known to have a lower binding affinity for the MHC class II molecule, failed to activate Vbeta5 T-cells whereas Vbeta22 T-cell activation was slightly decreased. Hence, early activation events highlighted the differential requirements of T cell families to respond to SEA. PMID- 14527510 TI - Prostaglandin E2 inhibits TNF production in murine bone marrow-derived dendritic cells. AB - Exposure to pathogens induces dendritic cells to release inflammatory cytokines and chemokines. The inflammatory response is controlled by endogenous agents such as anti-inflammatory cytokines, glucocorticoids, anti-inflammatory neuropeptides, and lipid mediators. This study is the first report on the inhibition by prostaglandin E2 (PGE2) of TNF release from bone marrow-derived dendritic cells stimulated with lipopolysaccharide (LPS), a TLR4 ligand, or peptidoglycan, a TLR2 ligand. The inhibition of TNF occurs at both mRNA and protein level. The inhibitory effect of PGE2 is mediated by the EP2 and EP4 receptors, and involves both PKA signaling and mediation by DC-derived IL-10. Intraperitoneal administration of PGE2 together with LPS results in a reduction in serum TNF and intracellular TNF in peritoneal exudate cells, compared to LPS alone. In addition, administration of PGE2 in vivo reduces the numbers of CD11c+ DCc that accumulate in the peritoneal cavity in response to LPS. The various implications of the PGE2-induced reduction in TNF are discussed. PMID- 14527511 TI - IL-2 and IL-15 manifest opposing effects on activation of nuclear factor of activated T cells. AB - IL-2 and IL-15 are cytokines involved in T cell activation and death. Their non shared receptors, IL-2Ralpha and IL-15Ralpha, are important in the homeostasis of lymphocytes as evidenced by gene deletion studies. How these cytokine/receptor systems affect T cell antigen receptor signaling pathways is poorly understood. Here, we show that the IL-2 and IL-15 cytokine/receptor alpha systems regulate activation of nuclear factor of activated T cells (NF-AT) in opposing ways. IL 15Ralpha increased while IL-2Ralpha decreased basal NF-AT activation status in a Jurkat transient transfection model. The effect of each of the alpha chain receptors on NF-AT activation was further opposed by addition of the respective cytokine. These effects were inhibited by anti-cytokine and anti-cytokine receptor reagents as well as by inhibitors of TCR signaling. These results suggest a novel pathway of cytokine action to regulate T cell signaling, activation, death, and homeostasis. PMID- 14527513 TI - Creation by mutagenesis of a mammalian Ca(2+) channel beta subunit that confers praziquantel sensitivity to a mammalian Ca(2+) channel. AB - Voltage-gated Ca(2+) channel beta subunits are important modulators of the pore forming alpha(1) subunit. We have cloned two schistosome beta subunits that confer sensitivity to the antischistosomal drug praziquantel (PZQ) to an otherwise insensitive mammalian alpha(1) subunit. The primary site of beta subunit interaction with alpha(1) subunits is the beta interaction domain (BID). The BID contains two conserved serines (225, 235 in rat beta2a) that constitute consensus sites for protein kinase C phosphorylation. However, these serines are absent in these schistosome beta subunits. Here we show that the capability to confer PZQ sensitivity can be created in the rat beta2a subunit by eliminating both serines in the BID. These results are consistent with, and should help our understanding of, the selective toxicity of PZQ. PMID- 14527512 TI - 4-1BB cross-linking enhances the survival and cell cycle progression of CD4 T lymphocytes. AB - 4-1BB, a T cell co-stimulatory receptor, prolongs the survival and multiplication of CD4 T cells. Cross-linking 4-1BB stimulated expression of the anti-apoptotic genes bcl-XL and bcl-2, as well as of cyclins D2 and E, and inhibited expression of the cyclin-dependent kinase (cdk) inhibitor p27kip1. Ova-activated CD4 T cells of 4-1BB-deficient/DO11.10 TCR transgenic mice survived less well and underwent less expansion than cells of wild type DO11.10 TCR transgenic mice. These findings demonstrate that 4-1BB is a co-stimulatory molecule for CD4 T cell survival and expansion in vivo. PMID- 14527514 TI - In vitro selection of high-affinity nucleic acid ligands to parasite target molecules. AB - The logic of using nucleic acids as pharmaceutical reagents is in part based on their capacity to interact with high affinity and specificity with other biological components. Considerable progress has been made over the past 10 years in the development of nucleic acid-based drug molecules using a variety of different technologies. One approach is a combinatorial technology that involves an iterative Darwinian-type in vitro evolution process, which has been termed SELEX for 'systematic evolution of ligands by exponential enrichment'. The procedure is a highly efficient method of identifying rare ligands from combinatorial nucleic acid libraries of very high complexity. It allows the selection of nucleic acid molecules with desired functions and it has been instrumental in the identification of a number of synthetic DNA and RNA molecules, so-called aptamers that recognise ligands of different chemical origin. The method is fast, it does not require special equipment and the selected aptamers typically bind their target with high affinity and high specificity. Here we summarise the recent examples of the SELEX technique within the context of identifying high-affinity ligands against parasite target molecules. PMID- 14527515 TI - Cleavage of trypanosome surface glycoproteins by alkaline trypsin-like enzyme(s) in the midgut of Glossina morsitans. AB - EP and GPEET procyclin, the major surface glycoproteins of procyclic forms of Trypanosoma brucei, are truncated by proteases in the midgut of the tsetse fly Glossina morsitans morsitans. We show that soluble extracts from the midguts of teneral flies contain trypsin-like enzymes that cleave the N-terminal domains from living culture-derived parasites. The same extract shows little activity against a variant surface glycoprotein on living bloodstream form T. brucei (MITat 1.2) and none against glutamic acid/alanine-rich protein, a major surface glycoprotein of Trypanosoma congolense insect forms although both these proteins contain potential trypsin cleavage sites. Gel filtration of tsetse midgut extract revealed three peaks of tryptic activity against procyclins. Trypsin alone would be sufficient to account for the cleavage of GPEET at a single arginine residue in the fly. In contrast, the processing of EP at multiple sites would require additional enzymes that might only be induced or activated during feeding or infection. Unexpectedly, the pH optima for both the procyclin cleavage reaction and digestion of the trypsin-specific synthetic substrate Chromozym-TRY were extremely alkaline (pH 10). Direct measurements were made of the pH within different compartments of the tsetse digestive tract. We conclude that the gut pH of teneral flies, from the proventriculus to the hindgut, is alkaline, in contradiction to previous measurements indicating that it was mildly acidic. When tsetse flies were analysed 48 h after their first bloodmeal, a pH gradient from the proventriculus (pH 10.6+/-0.6) to the posterior midgut (pH 7.9+/-0.4) was observed. PMID- 14527516 TI - The development of the macrogamete and oocyst wall in Eimeria maxima: immuno light and electron microscopy. AB - We have identified, and followed the development of three macrogamete organelles involved in the formation of the oocyst wall of Eimeria maxima. The first were small lucent vacuoles that cross-reacted with antibodies to the apple domains of the Toxoplasma gondii microneme protein 4. They appeared early in development and were secreted during macrogamete maturation to form an outer veil and were termed veil forming bodies. The second were the wall forming bodies type 1, large, electron dense vacuoles that stained positively only with antibodies raised to an enriched preparation of the native forms of 56 (gam56), 82 (gam82) and 230 kDa (gam230) gametocyte antigens (termed anti-APGA). The third were the wall forming bodies type 2, which appeared before the wall forming bodies type 1 but remain enclosed within the rough endoplasmic reticulum and stained positively with antibodies raised to recombinant versions of gam56 (anti-gam56), gam82 (anti gam82) and gam230 (anti-gam230) plus anti-APGA. At the initiation of oocyst wall formation, the anti-T. gondii microneme protein 4 positive outer veil detached from the surface. The outer layer of the oocyst wall was formed by the release of the contents of wall forming bodies type 1 at the surface to form an electron dense, anti-APGA positive layer. The wall forming bodies type 2 appeared, subsequently, to give rise to the electron lucent inner layer. Thus, oocyst wall formation in E. maxima represents a sequential release of the contents of the veil forming bodies, wall forming bodies types 1 and 2 and this may be controlled at the level of the rough endoplasmic reticulum/Golgi body. PMID- 14527517 TI - Characterisation of alpha-1 giardin: an immunodominant Giardia lamblia annexin with glycosaminoglycan-binding activity. AB - Alpha-1 giardin is an immunodominant protein in the intestinal protozoan parasite Giardia lamblia. The Triage((R)) parasite panel, used to detect copro-antigens in stool from giardiasis patients, reacts with an epitope between amino acids 160 and 200 in alpha-1 giardin. This region of the protein is also highly immunogenic during human infections. Alpha-1 giardin is related to annexins and like many other annexins it was shown to be plasma membrane associated. Immunoelectron and immunofluorescence microscopy revealed that some alpha-1 giardin are displayed on the surface of recently excysted cells. Recombinant alpha-1 giardin displayed a Ca(2+)-dependent binding to glycosaminoglycans (GAGs), in particular heparan sulphate, a common GAG in the intestinal tract. Recombinant alpha-1 giardin bound to thin sections of human small intestine, a binding which could be inhibited by adding increasing concentrations of sulphated sugars. A surface associated trypsin activated Giardia lectin (taglin) has been suggested to be important for G. lamblia attachment. In this study we show that a monoclonal antibody that inhibits taglin recognises alpha-1 and alpha-2 giardin. Thus, alpha-1 giardin is a highly immunoreactive GAG-binding protein, which may play a key role in the parasite-host interaction. Our results further show a conserved function of annexins from lower to higher eukaryotes. PMID- 14527518 TI - Parasite community structure within and across host populations of a marine pelagic fish: how repeatable is it? AB - The geographical variation in parasite community structure among populations of the same host species remains one of the least understood aspects of parasite community ecology. Why are parasite communities clearly structured in some host populations, and randomly assembled in others? Here, we address this fundamental question using data on the metazoan parasite communities of different host size classes of four distinct populations of a small pelagic fish, the Argentine anchovy, Engraulis anchoita, from the South West Atlantic. Within each fish sample, fish length was correlated with both the total intensity of parasites and species richness among infracommunities. More importantly, average fish length correlated with mean infracommunity richness and mean total intensity across the fish samples, indicating that the characteristics of parasite assemblages in a fish population are strongly influenced by the size of its fish in relation to those in other populations. Nested subset patterns were observed in about half of the fish samples. This means that the presence or absence of parasite species among fish individuals is often not random; however, no repeatability of nestedness among component communities was observed. Average fish length did not influence directly the likelihood that a parasite assemblage was significantly nested. However, variables influenced by average fish length, namely mean infracommunity richness and mean total intensity, determine the probability that a nested hierarchy will be observed; host size may thus indirectly affect parasite community structure either itself or via its influence on host movement and feeding patterns. To some extent, this apparent link may be due to the sensitivity of nestedness analyses to the proportion of presence in a presence/absence matrix; this in itself is a biological feature of the parasite community, however, which is associated with mean host length. PMID- 14527519 TI - Infection of human and bovine epithelial cells with Cryptosporidium andersoni induces apoptosis and disrupts tight junctional ZO-1: effects of epidermal growth factor. AB - The effects of Cryptosporidium andersoni on human or bovine epithelia are poorly defined. Epidermal growth factor inhibits colonisation of the gastrointestinal epithelium with bacteria and the enteric protozoan parasite Giardia lamblia. This study characterised whether C. andersoni infects human or bovine epithelial cells in vitro, assessed its impact on apoptosis and tight junctional Zonula-Occludens 1, and determined whether these effects may be altered by epidermal growth factor. Monolayers of human colonic CaCo(2) cells, SCBN (non-malignant small intestinal epithelial cells), and Madin Darby bovine kidney epithelial cell lines (MDBK and NBL-1) were grown to confluency in Dulbecco's Modified Eagle Medium. Monolayers were assigned to one of three experimental groups-(1) control: exposed to culture medium alone; (2) untreated: exposed to 10(3) live C. andersoni oocysts or (3) epidermal growth factor-treated: apically pre-treated with recombinant human epidermal growth factor and then exposed to Cryptosporidium. Oocyst viability, infection with Cryptosporidium, apoptosis, and integrity of tight junctional Zonula-Occludens-1 were assessed. In addition, live Cryptosporidium oocysts were incubated with epidermal growth factor to assess whether epidermal growth factor had cryptosporicidial activity. Cryptosporidium andersoni oocysts infected all human and bovine monolayers, increased nuclear fragmentation, and disrupted Zonula-Occludens-1. Apical epidermal growth factor significantly reduced infection with C. andersoni in all cell lines and inhibited the Cryptosporidium-induced apoptosis and disruption of Zonula-Occludens-1. Epidermal growth factor did not affect oocyst viability. PMID- 14527520 TI - Intradermal inoculations of low doses of Leishmania major and Leishmania amazonensis metacyclic promastigotes induce different immunoparasitic processes and status of protection in BALB/c mice. AB - In order to simulate the natural long term parasitisms which may occur in mammals infected with Leishmania, cutaneous leishmaniases due to Leishmania major or Leishmania amazonensis were induced using a model based on the inoculation of 10 1000 metacyclic promastigotes into the ear dermis of BALB/c mice. The final outcome of these parasitisms was dependent upon the number of inoculated parasites. Only some of the mice inoculated with ten parasites displayed cutaneous lesions, whereas most mice infected with 100 metacyclics and all mice infected with 1000 metacyclics developed progressive lesions. We found, using the latter experimental conditions, that the onset of the pathology was associated with: (a) parasite multiplication in the inoculation site and the draining lymph node correlating with an increase of the lymph node cell number, especially in L. major-infected mice; and (b) the detection of lymph node cells, at least in part CD4(+) T lymphocytes, able to produce high levels of interferon-gamma, interleukin (IL)-4, IL-10 and IL-13. Thereafter, mice infected by L. major harboured few parasites in the ear and had a 100-fold reduction in lymph node parasite load between 23 and 40 weeks post-inoculation. In contrast, the parasite loads of L. amazonensis-infected mice remained stable in the ear and increased in nodes during the same period of time. Only L. major-infected mice that exhibited cutaneous lesions in the primary site were resistant to the re-inoculation of 1000 metacyclic promastigotes, whereas all L. amazonensis-primary infected mice remained susceptible to a second homologous challenge. These results are the first to document that a status of resistance to re-infection, referred to concomitant immunity, is coupled to the development of primary progressive lesions in L. major-infected BALB/c mice. Such a protective status is absent in L. amazonensis-infected BALB/c mice. PMID- 14527522 TI - The mitochondrial genome of Strongyloides stercoralis (Nematoda) - idiosyncratic gene order and evolutionary implications. AB - The complete mitochondrial genome sequence of the parasitic nematode Strongyloides stercoralis was determined, and its organisation and structure compared with other nematodes for which complete mitochondrial sequence data were available. The mitochondrial genome of S. stercoralis is 13,758 bp in size and contains 36 genes (all transcribed in the clockwise direction) but lacks the atp8 gene. This genome has a high T content (55.9%) and a low C content (8.3%). Corresponding to this T content, there are 16 (poly-T) tracts of >/=12 Ts distributed across the genome. In protein-coding genes, the T bias is greatest (76.4%) at the third codon position compared with the first and second codon positions. Also, the C content is higher at the first (9.3%) and second (13.4%) codon positions than at the third (2%) position. These nucleotide biases have a significant effect on predicted codon usage patterns and, hence, on amino acid compositions of the mitochondrial proteins. Interestingly, six of the 12 protein coding genes are predicted to employ a unique initiation codon (TTT), which has not yet been reported for any other animal mitochondrial genome. The secondary structures predicted for the 22 transfer RNA (trn) genes and the two ribosomal RNA (rrn) genes are similar to those of other nematodes. In contrast, the gene arrangement in the mitochondrial genome of S. stercoralis is different from all other nematodes studied to date, revealing only a limited number of shared gene boundaries (atp6-nad2 and cox2-rrnL). Evolutionary analyses of mitochondrial nucleotide and amino acid sequence data sets for S. stercoralis and seven other nematodes demonstrate that the mitochondrial genome provides a rich source of phylogenetically informative characters. In conclusion, the S. stercoralis mitochondrial genome, with its unique gene order and characteristics, should provide a resource for comparative mitochondrial genomics and systematics studies of parasitic nematodes. PMID- 14527521 TI - Identification and molecular characterisation of DNA damaging agent induced expression of Plasmodium falciparum recombination protein PfRad51. AB - Rad51 protein, the eukaryotic homologue of Escherichia coli RecA protein plays a pivotal role in recombinational repair mechanism. We have identified a new homologue of Rad51 from the apicomplexan parasite Plasmodium falciparum, designated PfRad51. The PfRad51 gene codes for a 351 amino acid polypeptide with a predicted molecular mass of 38720, and shares 66-75% sequence identity within the catalytic region with Rad51 from human, yeast and other protozoan parasites such as Trypanosoma and Leishmania. The expression of PfRad51 transcript as well as protein in the intra-erythrocytic in vitro culture of P. flalciparum was found to be up-regulated in response to the DNA damaging agent methyl methanesulfonate, suggesting its functional involvement in recombinational repair process. PfRad51 is the first apicomplexan gene identified that codes for a recombination protein, and it offers an excellent model for studying DNA damage inducible gene expression in such parasites. PMID- 14527523 TI - The protozoan parasite, Theileria annulata, induces a distinct acute phase protein response in cattle that is associated with pathology. AB - Acute phase proteins (APP) are synthesised in the liver in response to the systemic presence of high levels of pro-inflammatory cytokines. Bacteria are considered to be strong inducers of APP whereas viruses are weak or non-inducers of APP. Very few reports have been published on APP induction by parasites. Here, we report that the tick-borne protozoan parasite of cattle, Theileria annulata, induced an atypical acute phase response in cattle. Following experimental infection, serum amyloid A (SAA) appeared first, followed by a rise in alpha(1) acid glycoprotein (alpha(1)AGP) in all animals, whereas haptoglobin, which is a major APP in cattle, only appeared in some of the animals, and generally at a low level. All three APP only became elevated around or after the appearance of schizonts in draining lymph nodes and after the first observed temperature rise. Increased alpha(1)AGP levels coincided with the appearance of piroplasms. The production of SAA and alpha(1)AGP correlated strongly with each other, and also with some clinical measures of disease severity including the time to fever, development of leucopaenia, parasitaemia and mortality. These results are consistent with the hypothesis that T. annulata causes severe pathology in susceptible cattle by inducing high levels of pro-inflammatory cytokines. PMID- 14527524 TI - Alterations in hexose, amino acid and peptide transporter expression in intestinal epithelial cells during Nippostrongylus brasiliensis infection in the rat. AB - Infection with the nematode Nippostrongylus brasiliensis induces various types of cytological alterations in the intestinal villus epithelium. The aim of this study was to analyse the expression of hexose, peptide and amino acid transporters in the small intestinal epithelium after infection. Brown-Norway rats were infected with 2000 N. brasiliensis L3 larvae and villus epithelial cells were isolated at various time points after infection. Expression of hexose transporters Na(+)/glucose cotransporter SGLT1 and glucose transporter GLUT-1, -2 and -5, a peptide transporter (PepT1) and an amino acid transporter (LAT2) was examined by reverse transcription-PCR, Western blotting or immunohistochemistry. Semi-quantitative reverse transcription-PCR studies of separated jejunal epithelial cells showed that expression levels of GLUT5, PepT1 and LAT2 were significantly decreased 7 and 14 days after infection, while these changes were not observed in the ileal epithelium. Although the apical surface glucose transporter SGLT1 showed no significant alteration in mRNA expression, Western blotting analyses of jejunal epithelial cell lysate showed a marked decrease. Contrary to SGLT1, GLUT5, PepT1 and LAT2, expression of GLUT1, which is essential in maintaining high rates of glucose influx, was significantly up-regulated in the jejunal epithelium 7 and 14 days after infection in reverse transcription-PCR as in Western blotting analyses. Immunohistochemical studies showed that GLUT1 immunoreactivity was localised to the basolateral membrane of intestinal epithelial cells 7 days after infection. These results show that N. brasiliensis infection results in an increase in GLUT1 and a decrease in various hexose, amino acid and peptide transporter expression in jejunal epithelial cells. Up regulation of GLUT1 might be a compensatory response in injured epithelial cells. PMID- 14527525 TI - A PCR-ELISA for the identification of cyathostomin fourth-stage larvae from clinical cases of larval cyathostominosis. AB - We report the use of six oligoprobes designed from intergenic spacer region sequences to identify fourth-stage larvae (L4) of the tribe Cyathostominae. Oligoprobes were designed for identification of the following species: Cylicocyclus ashworthi, Cylicocyclus nassatus, Cylicocyclus insigne, Cyathostomum catinatum, Cylicostephanus goldi, and Cylicostephanus longibursatus. A seventh probe was designed as a positive control to identify all these members of the Cyathostominae. The intergenic spacer region was amplified by PCR using conserved primers. Initially, three oligoprobes were used in Southern blot analysis. To facilitate high-throughput identification, these and a further four oligoprobes were developed for use in a PCR-ELISA. All probes were validated for their ability to detect cyathostomin PCR products in the PCR-ELISA, using DNA from morphologically identified adult parasites. Initially, 712 L4 were isolated from the diarrhoeic faeces from horses (n=17) with clinical larval cyathostominosis. PCR products from 522 of these L4 were subjected to analysis, with 413 L4 being identified as one of the aforementioned species. With reference to individual species analysis, 28.5% of the 522 L4 were identified as C. longibursatus, 25.7% as C. nassatus, 15.9% as C. ashworthi, 7.3% as C. goldi and 1.7% as C. catinatum. No L4 were identified as being C. insigne species. When L4 within faeces from individual horses were compared, no sample was found to comprise parasites of one species. The least number of species identified in a single sample was two. This study suggests that clinical larval cyathostominosis is predominantly caused by mixed-species infections. PMID- 14527526 TI - Effectiveness of an empirically supported treatment for social phobia in the field. AB - This study examined the effectiveness of individual exposure combined with cognitive restructuring for social phobia in a clinical setting as well as the influence of sample restriction criteria on the effect size. Participants were 217 unselected patients with a primary diagnosis of social phobia who were treated by 57 therapists in four outpatient clinics of the Christoph-Dornier Foundation of Clinical Psychology in Germany. Results 6 weeks after the end of therapy showed highly significant reductions in social phobic fears and avoidance as well as in general anxiety and symptoms of depression. However, patients who dropped out during therapy reported a significantly higher degree of depression. Results did not differ between the four outpatient clinics and are comparable with the average effect-sizes reported by meta-analytic studies of controlled efficacy research, using selected patients. Also, restricting the sample according to the selection criteria often applied in research settings did not result in higher effect sizes for the applied outcome measures. We conclude that individual cognitive behavioural therapy for social phobia can be transported from research settings to the field of mental health. PMID- 14527527 TI - Treatment of pharmacotherapy-refractory posttraumatic stress disorder among Cambodian refugees: a pilot study of combination treatment with cognitive behavior therapy vs sertraline alone. AB - Cambodian refugees with posttraumatic stress disorder (PTSD) represent a cohort in severe need of treatment, but little information is available to guide treatment choices. We selected a sample of pharmacotherapy-refractory individuals to test the efficacy of combination treatment with sertraline and cognitive behavior therapy (CBT) for treating PTSD. Participants in this pilot study were ten Khmer-speaking women who had been at a mean age of 22-26 years during the Pol Pot period (1975-1979). These patients were randomly assigned to either sertraline alone or combined treatment. We found that combined treatment offered additional benefit in the range of medium to large effect sizes for PTSD and associated symptoms. Our findings indicate that substantial gains can be achieved by adding CBT to pharmacotherapy for PTSD, and that a program of CBT emphasizing information, exposure, and cognitive-restructuring can be successfully modified for Khmer-speaking refugees. PMID- 14527528 TI - Fear information and the development of fears during childhood: effects on implicit fear responses and behavioural avoidance. AB - Field, Argyris and Knowles (Behav Res Ther 39 (2001) 1259), and Field, Hamilton, Knowles and Plews (Behav Res Thera 41 (2003) 113) have developed a prospective paradigm for testing Rachman's (Behav Res Ther 15 (1977) 375) proposition that fear information is important in the development of fears and phobias in children. Despite this paradigm being an advance on retrospective reports, the research so far has been restricted to self-reported fear beliefs measured after the information is given. This gives rise to two possible shortcomings: (1) the effects could simply reflect demand characteristics resulting from children conforming to the experimental demands, and (2) although fear information changes beliefs, this might not translate into the behavioural change that would be expected if this information has a powerful effect relevant to the development of pathological fear. This paper describes an experiment that attempts to address these concerns by improving Field et al.'s (2001, 2003) basic paradigm but with the addition of two measures: (1) a behavioural measure of avoidance, and (2) an implicit attitude task that should not be susceptible to deliberate attempts to conform to experimental demands. The result showed that negative and positive information have dramatic, and opposite, effects on self-reported fear beliefs, behavioural avoidance and implicit attitudes. There were no effects of gender on any of these results. This study fully supports Rachman's model and suggests that past work does not merely reflect demand characteristics and that fear information increases behavioural avoidance as well as fear beliefs. PMID- 14527529 TI - Pain coping and social support as predictors of long-term functional disability and pain in early rheumatoid arthritis. AB - Pain-related avoidance factors and social resources, as assessed by pain coping and social support, are supposed to have lasting effects on functional disability and pain in chronic pain disorders. As a follow-up to a prospective study demonstrating short-term effects after one year (Behaviour Research and Therapy, 36, 179-193, 1998), the role of pain coping and social support at the time of diagnosis was investigated in relationship to the long-term course of functional disability and pain after three and five years in 78 patients with rheumatoid arthritis (RA), taking into account personality characteristics of neuroticism and extraversion, clinical status and use of medication. In line with findings at the one-year follow-up, results showed that more passive pain coping predicted functional disability at the three-year, but not the five-year follow-up. In addition, low levels of social support at the time of diagnosis consistently predicted both functional disability and pain at the three and five-year follow ups. Results indicate that pain coping and social support, assessed very early in the disease process, can affect long-term functional disability and pain in RA, and suggest that early interventions focusing on pain-related avoidance factors and social resources for patients at risk may beneficially influence long-term outcomes in RA. PMID- 14527530 TI - Anxiety and selective attention in obsessive-compulsive disorder. AB - Recently, there has been increasing evidence for information-processing deficits in individuals with obsessive-compulsive disorder (OCD). While impairments in selective attention have been identified to be central to the symptomatology of OCD, the role that situational anxiety plays in attentional processes has not been fully explored. Previous research findings were limited to tasks containing anxiety-relevant materials, only permitting for the evaluation of the impact of anxiety on simultaneous cognitive processing. Furthermore, it has not yet been determined whether the impact of anxiety is limited to selective attention or is indicative of a more general cognitive impairment. This study was designed to examine the role that situational anxiety plays in selective attention impairments. OCD participants and controls were presented with an anxiety producing statement and a neutral statement, followed by the Stroop Task. Results indicated that situational anxiety plays a significant role in the performance of tasks that require selective attention in OCD. A significant deterioration was detected in performance on selective attention tasks for the OCD participants after confronting anxiety-provoking scenarios, as compared to neutral scenarios. Anxiety did not impair performance on simple reading tasks. Possible explanations are discussed. PMID- 14527531 TI - Attentional bias to threat in social phobia: facilitated processing of threat or difficulty disengaging attention from threat? AB - There is a growing body of research pointing to the possibility that anxious individuals may have difficulty disengaging their attention from threat-relevant information when this information is task irrelevant (e.g., Amir, N., & Elias, J. (2002). Allocation of attention to threat in social phobia: difficulty in disengaging from task irrelevant cues, Manuscript under review; The Quarterly J. Expo. Psycho. 54A (2001) 665). In the current paper, we report a direct test of this hypothesis in individuals with social phobia. Participants performed a variation of the Posner paradigm (Quart. J. Exp. Psycho. 32 (1980) 3). Social threat, neutral, or positive words cued one of two locations on the computer screen. After the cue disappeared, participants had to detect a probe ("(*)") that appeared in one of the two locations. On some trials the cue was valid (i.e., the probe appeared in the same location as the cue). On other trials the cue was invalid (the probe appeared in a different location than the cue). Yet, on other trials, no cue was presented. All participants were slower in detecting probes following invalid cues than probes following valid cues. Furthermore, individuals with social phobia showed significantly longer response latencies when detecting invalidly cued targets than did controls, but only when the probe followed a social threat word. These results suggest that individuals with social phobia may have difficulty disengaging their attention from socially threatening material. PMID- 14527532 TI - Active-imaginal exposure: examination of a new behavioral treatment for cynophobia (dog phobia). AB - The aims of this study were to investigate exposure-based treatments for cynophobia (dog phobia) and to test a newly developed hybrid imaginal exposure treatment that we have named active imaginal exposure. The treatment introduces an in vivo coping component to imaginal exposure whereby the patient physically performs coping responses to an imagined feared stimulus. Eighty-two participants meeting DSM-IV criteria for specific phobia (animal subtype) were randomly assigned to one of three 30-min. treatments: (a) active-imaginal exposure (AI), (b) imaginal exposure alone (IE), or (c) graduated in vivo exposure (IV). Participants completed a behavioral approach test at pre, post, and four-week follow-up. Significant pre- to posttreatment improvement was observed in all three treatment conditions. Response rates at posttreatment were 51.9, 62.1, and 73.1% for the IE, AI, and IV groups respectively. Likewise, effect sizes at posttreatment were 0.76, 1.41, and 1.55 for the IE, AI, and IV groups respectively. Although in the predicted direction, the between group differences were not significant. A similar pattern of results was observed at follow-up. Further, safety behavior utilization during treatment was associated with less improvement--particularly in the two imaginal treatment conditions. Exposure treatments of dog phobia appear feasible and effective in reducing phobic fear and avoidance associated with dog phobia. Furthermore, preliminary evidence suggests that our active-imaginal exposure treatment may be a viable alternative to in vivo exposure. PMID- 14527533 TI - Social anxiety and interpersonal perception: a social relations model analysis. AB - Cognitive models of social phobia posit that an individual's negative beliefs about the way he or she is perceived by others (metaperceptions) are a core feature of the disorder. The social relations model () was used to analyze interpersonal perception data collected following unstructured social interactions in 62 socially anxious (SA) and 62 not socially anxious (NSA) individuals. Using this model, the interpersonal perceptions were analyzed to evaluate whether pathological levels of social anxiety are associated with self perceptions, metaperceptions, and perceptions from others. SA participants saw themselves negatively and believed others saw them negatively. Although seen as more nervous by others, SA participants were not seen as less likeable. A mediational model demonstrated that the negative metaperceptions of SA individuals were more a function of their own self-perceptions than the negative perceptions of others. These findings were not attributable to depressive symptoms. Implications for theory and treatment of social phobia are discussed. PMID- 14527536 TI - Neuronal mechanisms for detection of motion in the field of view. AB - The visual system cannot rely only upon information from the retina to perceive object motion because identical retinal stimulations can be evoked by the movement of objects in the field of view as well as by the movements of retinal images self-evoked by eye movements. We clearly distinguish the two situations, perceiving object motion in the first case and stationarity in the second. The present work deals with the neuronal mechanisms that are likely involved in the detection of real motion. In monkeys, cells that are able to distinguish real from self-induced motion (real-motion cells) are distributed in several cortical areas of the dorsal visual stream. We suggest that the activity of these cells is responsible for motion perception, and hypothesize that these cells are the elements of a cortical network representing an internal map of a stable visual world. Supporting this view are the facts that: (i) the same cortical regions in humans are activated in brain imaging studies during perception of object motion; and (ii) lesions of these same regions produce selective impairments in motion detection, so that patients interpret any retinal image motion as object motion, even when they result from her/his eye movements. Among the areas of the dorsal visual stream rich in real-motion cells, V3A and V6, likely involved in the fast form and motion analyses needed for visual guidance of action, could use real motion signals to orient the animal's attention towards moving objects, and/or to help grasping them. Areas MT/V5, MST and 7a, known to be involved in the control of pursuit eye movements and in the analysis of visual signals evoked by slow ocular movements, could use real-motion signals to give a proper evaluation of motion during pursuits. PMID- 14527537 TI - Cells in monkey STS responsive to articulated body motions and consequent static posture: a case of implied motion? AB - We show that populations of visually responsive cells in the anterior part of the superior temporal sulcus (STSa) of the macaque monkey code for the sight of both specific articulated body actions and the consequent articulated static body postures. We define articulated actions as actions where one body part (e.g. a limb or head) moves with respect to the remainder of the body which remains static; conversely non-articulated actions are actions where the equivalent body parts do not move with respect to each other but move as one. Similarly, articulated static body postures contain a torsion or rotation between parts, while non-articulated postures do not. Cells were tested with the sight of articulated and non-articulated actions followed by the resultant articulated or non-articulated static body postures. In addition, the static body postures that formed the start and end of the actions were tested in isolation. The cells studied did not respond to the sight of non-articulated static posture, which formed the starting-point of the action, but responded vigorously to the articulated static posture that formed the end-point of the action. Other static postures resembling the articulated end-point posture, but which were in a more relaxed muscular state (i.e. non-articulated), did not evoke responses. The cells did not respond to body actions that were less often associated with the effective static articulated postures. Our results suggest that the cells' responses were related to the implied action rather than the static posture per se. We propose that the neural representations in STSa for actual biological motion may also extend to biological motion implied from static postures. These representations could play a role in producing the activity in the medial temporal/medial superior temporal (V5(MT)/MST) areas reported in fMRI studies when subjects view still photographs of people in action. PMID- 14527538 TI - Response latencies of neurons in visual areas MT and MST of monkeys with striate cortex lesions. AB - Cortical area, MT (middle temporal area) is specialized for the visual analysis of stimulus motion in the brain. It has been suggested [Brain 118 (1995) 1375] that motion signals reach area MT via two dissociable routes, namely a 'direct' route which bypasses primary visual cortex (area, striate cortex (V1)) and is specialized for processing 'fast' motion (defined as faster than 6 degrees/s) with a relatively short latency, and an 'indirect' route via area V1 for processing 'slow' motion (slower than 6 degrees/s) with a relatively long latency. We tested this proposal by measuring the effects of unilateral V1 lesions on the magnitudes and latencies of responses to fast- and slow-motion (depicted by random dot kinematograms (RDK) ) of single neurons in areas MT and medial superior temporal area (MST) of anaesthetized macaque monkeys. In the unlesioned hemisphere contralateral to a V1 lesion, response magnitudes and latencies of MT neurons were similar to those previously reported from MT neurons in normal monkeys, and there was no significant association between slow movement and long response latency (>100 ms), or between fast movement and short latency (< or =100 ms). V1 lesions led to diminished response magnitudes and increased latencies in area MT of the lesioned hemisphere, but did not selectively abolish MT responses to slow moving stimuli, or abolish long-latency responses to either slow- or fast-moving stimuli. Response magnitudes and latencies in area MST, which receives visual inputs directly from area MT and is also specialized for visual analysis of motion, were unaffected by V1 lesions (though we have shown elsewhere that directionally-selective responses in both areas were impaired by V1 lesions). Overall, the results are incompatible with the hypothesis that there are dissociable routes to MT specialized for processing separately fast and slow motion. PMID- 14527539 TI - Similarities and differences in motion processing between the human and macaque brain: evidence from fMRI. AB - The present report reviews a series of functional magnetic resonance imaging (fMRI) activation studies conducted in parallel in awake monkeys and humans using the same motion stimuli in both species. These studies reveal that motion stimuli engage largely similar cortical regions in the two species. These common regions include MT/V5 and its satellites, of which FST contributes more to the human motion complex than is generally assumed in human imaging. These results also establish a direct link between selectivity of MT/V5 neurons for speed gradients and functional activation of human MT/V5 by three-dimensional (3D) structure from motion stimuli. On the other hand, striking functional differences also emerged: in humans V3A and several regions in the intraparietal sulcus (IPS) are much more motion sensitive than their simian counterparts. PMID- 14527540 TI - Normal and anomalous development of visual motion processing: motion coherence and 'dorsal-stream vulnerability'. AB - Directional motion processing is a pervasive and functionally important feature of the visual system. Behavioural and VEP studies indicate that it appears as a cortical function after about 7 weeks of age, with global processing, motion based segmentation, and the use of motion in complex perceptual tasks emerging shortly afterwards. A distinct, subcortical motion system controls optokinetic nystagmus (OKN) from birth, showing characteristic monocular asymmetries which disappear as binocular cortical function takes over in normal development. Asymmetries in cortical responses are linked to this interaction in a way that is not yet fully understood. Beyond infancy, a range of developmental disorders show a deficit of global motion compared to global form processing which we argue reflects a general 'dorsal-stream vulnerability'. PMID- 14527541 TI - Visual motion sensitivity and reading. AB - Reading is more difficult than speaking because an arbitrary set of visual symbols must be rapidly identified, ordered and translated into the sounds they represent. Many poor readers have particular problems with the rapid visual processing required for these tasks because they have a mild impairment of the visual magnocellular system. This deficit has been demonstrated using neuropathological, evoked potential, functional magnetic resonance imaging and psychophysical techniques. The sensitivity of the M-system in both good and bad readers correlates with their orthographic abilities, suggesting that the M system plays an important part in their development. This role is probably to mediate steady direction of visual attention and eye fixations on words. Thus many children with reading difficulties have unsteady eye control and this causes the letters they are trying to read to appear to move around, so that they cannot tell what order they are meant to be in. Therefore, boosting M-performance using yellow filters, or training eye fixation, can improve reading performance very significantly. Several genetic linkage studies have associated reading difficulties with the MHC control region on the short arm of chromosome 6. This system has recently been shown to help regulate the differentiation of M-cells. This association could also explain the high incidence of autoimmune conditions in poor readers. Other chromosomal sites are associated with the metabolism of polyunsaturated fatty acids (PUFAs) as found in fish oils, and this could explain why PUFA supplements can improve reading. PMID- 14527542 TI - The effect of expectation on facilitation of colour/form conjunction tasks by TMS over area V5. AB - In an earlier paper, we reported task-specific impairments and improvements caused by applying TMS over cortical visual area V5 [Proceedings of the Royal Society of London B 265 (1998) 537]. The phenomenon is further investigated in the present study using two of the previous tasks: a motion/form conjunction in which TMS impaired performance and a colour/form conjunction in which performance was enhanced with TMS. In the earlier experiment, subjects were presented with blocks of trials of one task type perhaps allowing some of the observed effects to arise from knowing the type of stimulus to be discriminated. When blocks of trials consisted of randomly mixed moving/form and colour/form conjunction tasks, TMS over V5 still impaired target-present responses for the moving/form conjunction, but the facilitation seen for colour/form conjunction target-present responses disappeared. We suggest that the competitive inhibition postulated between visual movement areas and colour areas in the brain, in our previous paper, are subject to expectation or knowledge of forthcoming stimulus type. PMID- 14527543 TI - Modulation of moving phosphene thresholds by transcranial direct current stimulation of V1 in human. AB - Small moving sensations, so-called moving phosphenes are perceived, when V5, a visual area important for visual motion analysis, is stimulated by transcranial magnetic stimulation (TMS). However, it is still a matter of debate if only V5 takes part in movement perception or other visual areas are also involved in this process. In this study we tested the involvement of V1 in the perception of moving phosphenes by applying transcranial direct current stimulation (tDCS) to this area. tDCS is a non-invasive stimulation technique known to modulate cortical excitability in a polarity-specific manner. Moving and stationary phosphene thresholds (PT) were measured by TMS before, immediately after and 10, 20 and 30 min after the end of 10 min cathodal and anodal tDCS in nine healthy subjects. Reduced PTs were detected immediately and 10 min after the end of anodal tDCS while cathodal stimulation resulted in an opposite effect. Our results show that the excitability shifts induced by V1 stimulation can modulate moving phosphene perception. tDCS elicits transient, but yet reversible effects, thus presenting a promising tool for neuroplasticity research. PMID- 14527544 TI - Perception of biological motion in parietal patients. AB - Three unilateral parietal patients were tested on their perception of biological motion, a special case of form-from-motion. Two patients had the lesion in the right, and one in the left parietal area. All patients could easily perform a classical form-from-motion task [Neuron 32 (2001) 985], but they were severely impaired in a visual search task using biological motion sequences. In particular, the left parietal patient showed a more severe loss. He was unable to identify even a single item. Overall our patients seemed to perform differently from the classical motion-blind patients described in the literature [Visual Cognition 3 (1996) 363; Eur. J. Neurol. 9 (2002) 463; Visual Neurosci. 5 (1990) 353] whose lesions included the visual cortical area V5. Since our patients' low level motion mechanisms are preserved, we suggest that the perception of biological motion relies on a high-level description of dynamic patterns [Cognition 80 (2001) 47], a mechanism that is impaired in parietal lobe patients. We discuss our results at the light of the recent theories suggesting that biological motion is performed by visual associative areas outside the classical motion pathways and that it is an active process dependent on attentional resources [Cognition 80 (2001) 47]. PMID- 14527545 TI - Can spatial and temporal motion integration compensate for deficits in local motion mechanisms? AB - We studied the motion perception of a patient, AMG, who had a lesion in the left occipital lobe centered on visual areas V3 and V3A, with involvement of underlying white matter. As shown by a variety of psychophysical tests involving her perception of motion, the patient was impaired at motion discriminations that involved the detection of small displacements of random-dot displays, including local speed discrimination. However, she was unimpaired on tests that required spatial and temporal integration of moving displays, such as motion coherence. The results indicate that she had a specific impairment of the computation of local but not global motion and that she could not integrate motion information across different spatial scales. Such a specific impairment has not been reported before. PMID- 14527546 TI - Attentional capture by colour and motion in cerebral achromatopsia. AB - Cerebral achromatopsia is a rare condition in which damage to the ventromedial occipital area of the cortex results in the loss of colour experience. Nevertheless, cortically colour-blind patients can still use wavelength variation to perceive form and motion. In a series of six experiments we examined whether colour could also direct exogenous attention in an achromatopsic observer. We employed the colour singleton paradigm, the phi motion effect, and the correspondence process to assess attentional modulation. Although colour singletons failed to capture attention, a motion signal, based solely on chromatic information, was able to direct attention in the patient. We then show that the effect is abolished when the chromatic contours of stimuli are masked with simultaneous luminance contrast. We argue that the motion effect is dependent on chromatic contrast mediated via intact colour-opponent mechanisms. The results are taken as further evidence for the processing of wavelength variation in achromatopsia despite the absence of colour experience. PMID- 14527547 TI - Multisensory contributions to the perception of motion. AB - The ability to process motion is crucial for coherent perception and action. While the majority of studies have focused on the unimodal factors that influence motion perception (see, for example, the other chapters in this Special Issue), some researchers have also investigated the extent to which information presented in one sensory modality can affect the perception of motion for stimuli presented in another modality. Although early studies often gave rise to mixed results, the development of increasingly sophisticated psychophysical paradigms are now enabling researchers to determine the spatiotemporal constraints on multisensory interactions in the perception of motion. Recent findings indicate that these interactions stand over-and-above the multisensory interactions documented previously for static stimuli, such as the oft-cited 'ventriloquism' effect. Neuroimaging and neuropsychological studies are also beginning to elucidate the network of neural structures responsible for the processing of motion information in the different sensory modalities, an important first step that will ultimately lead to the determination of the neural substrates underlying these multisensory contributions to motion perception. PMID- 14527549 TI - Solution-phase synthesis of novel delta2-isoxazoline libraries via 1,3-dipolar cycloaddition and their antifungal properties. AB - The synthesis of novel imidazolyl substituted delta2-isoxazoline libraries are currently of high interest. We report here in the full details of a study leading to the synthesis and antifungal activities of 3-(-2-butyl-4-chloro-1H-imidazolyl) substituted delta2-isoxazolines. The solution phase synthesis of the title compounds was accomplished via 1,3-dipolar cycloaddition of in situ generated nitryl oxides from aldoximes with mono substituted alkenes to obtain the compound libraries contain an imidazole functionality in addition to the isoxazoline rings. The newly synthesized compounds when tested in vitro in solid agar culture exerted a potent antifungal activity against Aspergillus flavus, Fusarium moniliforme and Botrydiplodia theobromae also MIC values were determined. The title 5-substituted-3-imidazolyl-delta2-isoxazoline compounds represent a novel class of potent antifungal agents. PMID- 14527550 TI - Anti-tumor activity of the farnesyl-protein transferase inhibitors arteminolides, isolated from Artemisa. AB - Members of the Artemisia genus are important medicinal plants found throughout the world. Arteminolides A-D (1-4), isolated from the aerial parts of Artemisia, have an inhibitory activity on farnesyl-protein transferase (FPTase; EC 2.5.1.29) in in vitro assay. This study was carried out with the purpose of validating anti tumor effects of the compounds in human tumor cells and mouse xenograft model. The arteminolides inhibited tumor cell growth in a dose-dependent manner. Furthermore, arteminolide C (3) blocked in vivo growth of human colon and lung tumor xenograft without the loss of body weight in nude mice. PMID- 14527551 TI - One-electron reduction characteristics of N(3)-substituted 5-fluorodeoxyuridines synthesized as radiation-activated prodrugs. AB - We designed and synthesized N(3)-substituted 5-fluorodeoxyuridines as radiation activated prodrugs of the antitumor agent, 5-fluorodeoxyuridine (5-FdUrd). A series of 5-FdUrd derivatives possessing a 2-oxoalkyl group at the N(3)-position released 5-FdUrd in good yield via one-electron reduction initiated by hypoxic irradiation. Cytotoxicity of the 5-FdUrd derivative possessing the 2 oxocyclopentyl group (3d) was low, but was enhanced by hypoxic irradiation resulting in 5-FdUrd release. PMID- 14527552 TI - 8-quinolinamines and their pro prodrug conjugates as potent blood-schizontocidal antimalarial agents. AB - Synthesis and antimalarial activities of N8-(4-amino-1-methylbutyl)-5-alkoxy-4 ethyl-6-methoxy-8-quinolinamines (5) and their pro prodrug analogues (6-7) prepared by covalently linking 5 to the redox-sensitive (8) and esterase sensitive (9) linkers through the amide linkage are reported. The most effective 8-quinolinamines [5c (R=C5H11) and 5f (R=C8H17)] have exhibited in vitro and in vivo biological efficacy superior to that of the standard drug chloroquine against both drug-sensitive and drug-resistant malaria strains. Analogues 6-7 were evaluated for in vivo blood-schizontocidal activity as potential pro prodrug models for the primary amino group containing 8-quinolinamines (5). The most effective pro prodrug analogue (6c) has displayed promising activities against drug-sensitive and drug-resistant strains of Plasmodia in vivo. PMID- 14527553 TI - Piericidins C5 and C6: new 4-pyridinol compounds produced by Streptomyces sp. and Nocardioides sp. AB - Piericidins C5 (1) and C6 (2), two new members of the piericidin family, were isolated from a Streptomyces sp. and a Nocardioides sp., together with known piericidins C1 (3), C2 (4), C3 (5), C4 (6), D1 (7), and A3 (8). The structures were determined on the basis of their spectroscopic data. Both new compounds inhibited cell division of fertilized starfish (Asterina pectinifera) eggs at the minimum inhibitory concentration of 0.09 microg/mL. PMID- 14527554 TI - Antifungal diterpenoids of Pseudolarix kaempferi, and their structure-activity relationship study. AB - The in vitro antifungal activities of 19 structurally diversified analogues of pseudolaric acids tested against the major pathogenic fungus Candida albicans has led to the establishment of a very clear structure-activity relationship of pseudolaric acids derivatives. Pseudolaric acid A was first found to be a potent antifungal component comparable with pseudolaric acid B. Among the tested 19 diterpenoids, pseudolaric acids A2 (1), B2 (3), B3 (4) and methyl pseudolarate A2 (2) are new isolates of the root bark of Pseudolarix kaempferi, and their structures were elucidated mainly by 2D-NMR techniques and chemical methods. Compounds 12-19 were first semi-synthesized by efficient routines from pseudolaric acid B. PMID- 14527555 TI - The 3D-QSAR study of antitumor arylsulfonylimidazolidinone derivatives by CoMFA and CoMSIA. AB - Three-dimensional quantitative structure-activity relationship (3D-QSAR) studies for a series of arylsulfonylimidazolidinone derivatives having antitumor activity were conducted using comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA). The in vitro cytotoxicity against human lung carcinoma (A549) exhibited a strong correlation with steric and electrostatic factors of the molecules. Four different types of models have been built using CoMFA and CoMSIA method with AM1 charge or Gasteiger-Huckel charge. By comparison of the statistical results of these models, model I obtained by CoMFA study with AM1 showed the best predictability of the antitumor activities based on the cross-validated value (0.642), conventional r2 (0.981), standard error of estimate (0.106) and PRESS value (0.170). PMID- 14527556 TI - Synthesis and cytotoxic activity of some new azapyranoxanthenone aminoderivatives. AB - A series of novel azapyranoxanthenones, bearing structural similarity to the acridone alkaloid acronycine have been designed and synthesized. Their in vitro cytotoxicities against the murine L1210 leukemia and the human solid tumor HT-29 cell lines have been investigated. The new derivatives exhibited interesting cytotoxic activity and were more potent than the parent compound. PMID- 14527558 TI - Synthesis and antiparasitic activity of albendazole and mebendazole analogues. AB - Albendazole (Abz) and Mebendazole (Mbz) analogues have been synthesized and in vitro tested against the protozoa Giardia lamblia, Trichomonas vaginalis and the helminths Trichinella spiralis and Caenorhabditis elegans. Results indicate that compounds 4a, 4b (Abz analogues), 12b and 20 (Mbz analogues) are as active as antiprotozoal agents as Metronidazole against G. lamblia. Compound 9 was 58 times more active than Abz against T. vaginalis. Compounds 8 and 4a also shown high activity against this protozoan. Compounds 4b and 5a were as active as Abz. None of the Mbz analogues showed activity against T. vaginalis. The anthelmintic activity presented by these compounds was poor. PMID- 14527557 TI - A system of protein target sequences for anti-RNA-viral chemotherapy by a vitamin B6-derived zinc-chelating trioxa-adamantane-triol. AB - The synthesis of the structurally unusual heterotricyclic compound 1-[3-hydroxy-5 (hydroxymethyl)-2-methyl-4-pyridinyl]-2,8,9-trioxaadamantane-3,5,7-triol (trivially named bananin, BN) from pyridoxylidenephloroglucinol and a theoretical prospect on possible biological activities of BN are presented in this report. Pyridoxylidenephloroglucinol is synthesized by Knoevenagel condensation of the vitamin B6 aldehyde pyridoxal with phloroglucinol. Pyridoxylidenephloroglucinol rearranges to light-yellow (4'RS)-1',4'-dihydrobananin by refluxing in 5M hydrochloric acid. Air oxidation subsequently forms BN in the heat which immediately yields orange-yellow (4'RS)-4'-chloro-1',4'-dihydrobananin by 1,4 addition of hydrogen chloride. This intermediate could be isolated but, interestingly, not a BN hydrochloride. Brown BN is finally achieved by base catalyzed elimination of hydrogen chloride from (4'RS)-4'-chloro-1',4' dihydrobananin. Regarding possible biological activities, it was demonstrated that BN acts as zinc (Zn2+) chelator. Therefore, a target of interest could be the human immunodeficiency virus type 1 (HIV-1) zinc finger HIV-1 RNA-binding nucleocapsid protein p7 (NCp7). Through suggested zinc ejection from HIV-1 genomic RNA psi-element-binding and HIV-1-RNA-duplex packaging NCp7 by BN, thus rendering NCp7 functionally obsolete, it is deduced that HIV-1 replication and effective infectious virion encapsidation could be inhibited by BN. Furthermore, theoretical and structural considerations propose that BN is converted into bananin 5'-monophosphate (BNP) by the cell type-ubiquitous human enzyme pyridoxal kinase (EC 2.7.1.35). Together with the putative antilentiviral retinoid vitamin A-vitamin B6 conjugate analogue B6RA (Kesel, A. J. Biochem. Biophys. Res. Comm. 2003, 300, 793), BNP is postulated to serve as effector in a system of protein target sequences RX(D/E) of RNA virus components. Human immunodeficiency Retroviridae (HIVs) could possibly be influenced by B6RA and BNP. In addition, candidate targets of B6RA and BNP could be adsorption, transcription and/or viral RNA replication of an interestingly wide RNA virus selection including Picornaviridae (poliovirus, human coxsackievirus, hepatitis A virus), Flaviviridae (yellow fever virus, Dengue virus, West Nile virus, Kunjin virus, St. Louis encephalitis virus, hepatitis C virus), Togaviridae (rubella virus), Coronaviridae (human coronavirus, human SARS-associated coronavirus), Rhabdoviridae (rabies virus), Paramyxoviridae (human parainfluenza virus, measles virus, human respiratory syncytial virus), Filoviridae (Marburg virus, Ebola virus), Bornaviridae (Borna disease virus), Bunyaviridae (Hantaan virus), Arenaviridae (Lassa virus), and Reoviridae (human rotavirus). The postulated scope of 'metabolically trapped' BNP might resemble the antiviral spectrum of the RNA-viral virustatic ribavirin. PMID- 14527559 TI - Design, synthesis and glutathione peroxidase-like properties of ovothiol-derived diselenides. AB - Eleven imidazole diselenides derived from the naturally occurring family of antioxidants, the ovothiols, were synthetised by cyclisation of selenoamides with trimethylsilyltrifluoromethanesulfonate or refluxing of cyanoamines in a selenium/sodium borohydride mixture. These compounds were assayed for their glutathione peroxidase-like (GSH Px-like) activity and their capacity to be reduced by glutathione. The most active molecules of the series were 4 times more potent in the GSH Px-like test than the widely known reference compound, ebselen. This catalytic activity was mediated by the formation of the antioxidant selenol intermediate upon partial but significant exchange reaction with glutathione. PMID- 14527560 TI - Inhibition of bovine plasma amine oxidase by 1,4-diamino-2-butenes and -2 butynes. AB - Bovine plasma amine oxidase (BPAO) was previously shown to be irreversibly inhibited by propargylamine and 2-chloroallylamine. 1,4-Diamine versions of these two compounds are here shown to be highly potent inactivators, with IC50 values near 20 microM. Mono-N-alkylation or N,N-dialkylation greatly lowered the inactivation potency in every case, whereas the mono-N-acyl derivatives were also weaker inhibitors and enzyme activity was recoverable. The finding that the bis primary amines 1,4-diamino-2-butyne (a known potent inhibitor of diamine oxidases) and Z-2-chloro-1,4-diamino-2-butene are potent inactivators of BPAO is suggestive of unexpected similarities between plasma amine oxidase and the diamine oxidases and implies that it may be unwise to attempt to develop selective inhibitors of diamine oxidase using a diamine construct. PMID- 14527561 TI - Design of EGFR kinase inhibitors: a ligand-based approach and its confirmation with structure-based studies. AB - Three-dimensional quantitative structure-activity relationship (3D-QSAR) models were developed for 100 anilinoquinazolines, inhibiting epidermal growth factor receptor (EGFR) kinase. The studies included molecular field analysis (MFA) and receptor surface analysis (RSA). The cross-validated r2 (r2cv) values are 0.81 and 0.79 for MFA and RSA, respectively. The predictive ability of these models was validated by 28 test set molecules. The results of the best QSAR model were further compared with structure-based investigations using docking studies with the crystal structure of EGFR kinase domain. The results helped to understand the nature of substituents at the 6- and 7-positions, thereby providing new guidelines for the design of novel inhibitors. PMID- 14527562 TI - Thio-derived disulfides as potent inhibitors of botulinum neurotoxin type B: implications for zinc interaction. AB - Botulinum neurotoxin type B causes the inhibition of acetylcholine release at the neuromuscular junction resulting in a flaccid paralysis designated botulism. This occurs through the cleavage of synaptobrevin, an intracellular critical component of neurotransmitter exocytosis, by the zinc-metallopeptidase activity of the smallest subunit of the toxin. Blocking the proteolytic activity may present an attractive approach to treat botulism as to date there is no efficient specific drug therapy available. We have therefore recently described a series of beta amino-thiol derived pseudotripeptides able of inhibiting the toxin at low (10(-8) M) concentration. In this study, binding characteristics of the protein's active site are explored through various structural modifications of the thiol functionality which was supposed to be a key structural constituent for effective zinc-ion chelation. Surprisingly, sulfanyl-derivatives such as symmetric disulfides were shown to be better inhibitors than their thiol-counterparts, the most potent compound displaying a Ki value of 3.4 nM. PMID- 14527563 TI - On how the conformation of biliverdins influences their reduction to bilirubins: a biological and molecular modeling study. AB - The cyclic 2,18-bridged biliverdin (2) is excreted in rat bile without reduction to the corresponding bilirubin. Conformational analysis, employing an optimized Monte Carlo method and a mixed Monte Carlo/stochastic dynamics, reveals that biliverdin IXalpha (1) and the cyclic analogue 2 adopt 'lock washer' conformations, stabilized by the presence of intramolecular hydrogen bonds between N23...H22N and, to a lesser extent, between N23...H24N. Although 2 is very similar in overall shape to 1, the former adopts a 'locked lock washer' conformation unable to undergo fluctuations, thus possibly hampering a proper recognition by biliverdin reductase. PMID- 14527564 TI - Antisense phosphorothioate oligodeoxyribonucleotide targeted against ICAM-1: synthetic and biological characterization of a process-related impurity formed during oligonucleotide synthesis. AB - A phosphorothioate-linked oligonucleotide bearing a 3'-terminal phosphorothioate monoester has been synthesized and characterized. This oligonucleotide has been identified as a process-related impurity formed during synthesis of ISIS 2302. Biological properties of the compound have been determined. Based on these data, it can be concluded that this species (3'-TPT) has biological properties similar to parent drug. PMID- 14527565 TI - Changes in brain activation patterns associated with learning of Korean words by Japanese: an fMRI study. AB - We used functional magnetic resonance imaging (fMRI) to explore the change in brain activation associated with the learning of Korean words written in Han-gul characters (K-words) by young Japanese at two stages. Subjects were 12 right handed native Japanese without previous knowledge of Korean words and characters. On the first day they were taught the pronunciation and meaning of 20 K-words. Then, after the first fMRI session (on day 2), they were given a set of 20 cards with the words and corresponding photographs. They also received a tape and were instructed to memorize the 20 K-words by studying them every day until the day of the second fMRI session (day 16). During the fMRI sessions, 20 Japanese words written in kana syllabograms (J-words) and the 20 previously presented K-words, as well as 20 new K-words (Kn-words) were presented visually for silent reading. The first J-word reading, relative to the first K-word reading, showed activation in the left angular gyrus. K-word reading relative to J-word reading during both sessions showed activation in occipital regions. Within these activated areas, session by condition interaction was found only in the left angular gyrus. The interaction between session and condition resulted from the fact that the differences in blood oxygenation-level-dependent signals between K-words and J words and between Kn-words and J-words were significantly greater in the first session than in the second session. From the results, we concluded that patterns of brain activation changed as the memory of the 20 K-words became fixed through daily practice and that reading of both Korean words and Japanese syllabograms engaged the left angular gyrus. PMID- 14527566 TI - The cerebellum and olfaction in the aging brain: a functional magnetic resonance imaging study. AB - The present study investigated activation of distinct cerebellar regions as a result of olfactory stimulation in healthy young and elderly adults using functional magnetic resonance imaging (fMRI). Ten young and 10 elderly adults were imaged using a 1.5-TMR scanner. The odorant amyl acetate was delivered in 12 s on, 40-s off cycles. Throughout the scan participants responded with a button press at first detection of each stimulus interval followed by a second button press upon odorant extinction. Images were processed with AFNI software. Elderly participants showed significantly decreased cerebellar activation in both the superior semilunar lobule (Crus I) and the inferior semilunar lobule (Crus II), two of three previously identified regions of interest for odor processing, compared to young adults. Interestingly, both groups showed similar levels of activation in the third region of interest, the posterior quadrangular lobule (VI), although the elderly showed more variability than the young did. Previous research identifying this area to be involved in attention may reflect the possibility that elderly adults engaged in olfactory tasks may show more variability than young participants in the degree of attentional demands needed for these tasks as a result of decreased olfactory abilities. PMID- 14527567 TI - A longitudinal study of brain morphometrics using quantitative magnetic resonance imaging and difference image analysis. AB - Serial quantitative magnetic resonance imaging (MRI) allows the detection of subtle volumetric changes in brain volume. We used serial volumetry and voxel based difference image analysis to quantify and characterize longitudinal changes in the hippocampus, cerebellum, and neocortex in younger and middle-age individuals. Paired volumetric MRI brain scans 3.5 years apart were performed on 90 healthy subjects 14 to 77 years old. Quantitative assessment of registered images included hippocampal volumetry, cerebellar volumetry, and automatically determined regional brain volumes. Longitudinal volume changes in three age epochs (<35, 35-54, >54 years) were compared and neocortical changes beyond regions of interest were visualized using filtered difference images. Cross sectional analysis revealed a significant association between age and reduction in all brain volumes except hippocampal volume. Changes in normalized hippocampal and white matter volume were significantly different among the three groups. Individual analysis revealed 5 subjects with significant longitudinal volume changes lying outside the normative range. Difference image analysis showed global involutional changes in the >54 age group. Our findings suggest that cross sectional observations in intracranial volume, cerebellar volume, and gray matter volume are likely to reflect uniform rates of volume loss or secular changes. Accelerated brain atrophy was seen from the age of 35-54 and increased rates of hippocampal atrophy from the age of 54. Our findings emphasize the importance of controlling for age effects when studying pathological brain changes over a wide age range. PMID- 14527568 TI - Dynamical components analysis of fMRI data through kernel PCA. AB - In parallel with standard model-based methods for the analysis of fMRI data, exploratory methods--such as PCA, ICA, and clustering--have been developed to give an account of the dataset with minimal priors: no assumption is made on the data content itself, but the data structure is assumed to show some properties (decorrelation, independence) that allow for the detection of structures of interest. In this paper, we present an alternative that tries to take into account some relevant knowledge for the analysis of the dataset, e.g., the experimental paradigm, while keeping the flexibility of exploratory methods: we use a prior temporal modeling of the data that characterizes each voxel time course. Two implementations are proposed: one based on the General Linear Model, the other one on more flexible short-term predictors, whose complexity is controlled by a Minimum Description Length approach. However, our main concern here is the construction of a multivariate model; the latter is performed with the help of a kernel PCA method that builds a redundant representation of the data through the nonlinearity of the kernel. This allows for a refinement in the description of the (temporal) patterns of interest. In particular, this helps in the characterization of subtle variations in the response to different experimental conditions. We illustrate the usefulness of nonlinearity through the analysis of a synthetic dataset and show on a real dataset how it helps to interpret the experimental results. PMID- 14527569 TI - The Cerefy Neuroradiology Atlas: a Talairach-Tournoux atlas-based tool for analysis of neuroimages available over the internet. AB - The article introduces an atlas-assisted method and a tool called the Cerefy Neuroradiology Atlas (CNA), available over the Internet for neuroradiology and human brain mapping. The CNA contains an enhanced, extended, and fully segmented and labeled electronic version of the Talairach-Tournoux brain atlas, including parcelated gyri and Brodmann's areas. To our best knowledge, this is the first online, publicly available application with the Talairach-Tournoux atlas. The process of atlas-assisted neuroimage analysis is done in five steps: image data loading, Talairach landmark setting, atlas normalization, image data exploration and analysis, and result saving. Neuroimage analysis is supported by a near-real time, atlas-to-data warping based on the Talairach transformation. The CNA runs on multiple platforms; is able to process simultaneously multiple anatomical and functional data sets; and provides functions for a rapid atlas-to-data registration, interactive structure labeling and annotating, and mensuration. It is also empowered with several unique features, including interactive atlas warping facilitating fine tuning of atlas-to-data fit, navigation on the triplanar formed by the image data and the atlas, multiple-images-in-one display with interactive atlas-anatomy-function blending, multiple label display, and saving of labeled and annotated image data. The CNA is useful for fast atlas assisted analysis of neuroimage data sets. It increases accuracy and reduces time in localization analysis of activation regions; facilitates to communicate the information on the interpreted scans from the neuroradiologist to other clinicians and medical students; increases the neuroradiologist's confidence in terms of anatomy and spatial relationships; and serves as a user-friendly, public domain tool for neuroeducation. At present, more than 700 users from five continents have subscribed to the CNA. PMID- 14527570 TI - Cholinergic enhancement modulates neural correlates of selective attention and emotional processing. AB - Neocortical cholinergic afferents are proposed to influence both selective attention and emotional processing. In a study of healthy adults we used event related fMRI while orthogonally manipulating attention and emotionality to examine regions showing effects of cholinergic modulation by the anticholinesterase physostigmine. Either face or house pictures appeared at task relevant locations, with the alternative picture type at irrelevant locations. Faces had either neutral or fearful expressions. Physostigmine increased relative activity within the anterior fusiform gyrus for faces at attended, versus unattended, locations, but decreased relative activity within the posterolateral occipital cortex for houses in attended, versus unattended, locations. A similar pattern of regional differences in the effect of physostigmine on cue-evoked responses was also present in the absence of stimuli. Cholinergic enhancement augmented the relative neuronal response within the middle fusiform gyrus to fearful faces, whether at attended or unattended locations. By contrast, physostigmine influenced responses in the orbitofrontal, intraparietal and cingulate cortices to fearful faces when faces occupied task-irrelevant locations. These findings suggest that acetylcholine may modulate both selective attention and emotional processes through independent, region-specific effects within the extrastriate cortex. Furthermore, cholinergic inputs to the frontoparietal cortex may influence the allocation of attention to emotional information. PMID- 14527571 TI - Brain changes after learning to read and play music. AB - Musically naive participants were scanned before and after a period of 15 weeks during which they were taught to read music and play the keyboard. When participants played melodies from musical notation after training, activation was seen in a cluster of voxels within the bilateral superior parietal cortex. A subset of these voxels were activated in a second experiment in which musical notation was present, but irrelevant for task performance. These activations suggest that music reading involves the automatic sensorimotor translation of a spatial code (written music) into a series of motor responses (keypresses). PMID- 14527572 TI - Common and distinct neural responses during direct and incidental processing of multiple facial emotions. AB - Whether common or distinct neural systems underpin perception of different emotions and the degree to which these systems are automatically engaged during emotional perception are unresolved. We performed an event-related fMRI experiment in which subjects viewed morphed emotional faces displaying low or high intensities of disgust, fear, happiness, or sadness under two task conditions. The amygdala and fusiform cortex responded to high-intensity expressions of all emotions, independent of task. Right superior temporal sulcus showed an additive effect of the emotion-directed task and high-intensity emotion. Ventromedial prefrontal and somatosensory cortices, regions implicated in providing representations of somatic states, showed enhanced activity during explicit emotional judgments. We failed to find predicted differences between emotions. The results suggest that amygdala contributes to task-independent perceptual processing of a range of emotions. We interpret ventromedial prefrontal and somatosensory cortex activations as evidence that these regions contribute to explicit emotion processing through linking emotion perception with representations of somatic states previously engendered by emotions. PMID- 14527573 TI - Attentional modulation of oscillatory activity in human visual cortex. AB - The effects of attentional modulation on activity within the human visual cortex were investigated using magnetoencephalography. Chromatic sinusoidal stimuli were used to evoke activity from the occipital cortex, with attention directed either toward or away from the stimulus using a bar-orientation judgment task. For five observers, global magnetic field power was plotted as a function of time from stimulus onset. The major peak of each function occurred at about 120 ms latency and was well modeled by a current dipole near the calcarine sulcus. Independent component analysis (ICA) on the non-averaged data for each observer also revealed one component of calcarine origin, the location of which matched that of the dipolar source determined from the averaged data. For two observers, ICA revealed a second component near the parieto-occipital sulcus. Although no effects of attention were evident using standard averaging procedures, time-varying spectral analyses of single trials revealed that the main effect of attention was to alter the level of oscillatory activity. Most notably, a sustained increase in alpha band (7-12 Hz) activity of both calcarine and parieto-occipital origin was evident. In addition, calcarine activity in the range of 13-21 Hz was enhanced, while calcarine activity in the range of 5-6 Hz was reduced. Our results are consistent with the hypothesis that attentional modulation affects neural processing within the calcarine and parieto-occipital cortex by altering the amplitude of alpha-band activity and other natural brain rhythms. PMID- 14527574 TI - Use of concurrent pupil dilation assessment to inform interpretation and analysis of fMRI data. AB - Potential contributions of concurrently acquired pupil dilation data to functional magnetic resonance imaging (fMRI) experiments were examined. Sixteen healthy participants completed a working memory task (digit sorting) during measurement of pupil dilation outside the fMRI environment and during concurrent 3T fMRI assessment. Pupil dilation increased parametrically with task difficulty inside and outside the scanner, on a similar time course, suggesting that task demand was similar in both environments. The time course of pupil dilation during digit sorting was similar to the time course of the fMRI signal in the middle frontal gyrus, suggesting that middle-frontal gyrus activity indexed the engagement working memory processes. Incorporating individual differences in pupil dilation improved the sensitivity and specificity of general linear modeling analyses of activity in the middle frontal gyrus, above and beyond standard analytic techniques. Results suggest concurrent pupil dilation during fMRI assessment can help to (1) specify whether task demand is the same inside and outside the fMRI environment, (2) resolve the extent to which fMRI signals reflect different aspects of event-related designs, and (3) explain variation in fMRI data due to individual differences in information processing. PMID- 14527575 TI - Learned material content and acquisition level modulate cerebral reactivation during posttraining rapid-eye-movements sleep. AB - We have previously shown that several brain areas are activated both during sequence learning at wake and during subsequent rapid-eye-movements (REM) sleep (Nat. Neurosci. 3 (2000) 831-836), suggesting that REM sleep participates in the reprocessing of recent memory traces in humans. However, the nature of the reprocessed information remains open. Here, we show that regional cerebral reactivation during posttraining REM sleep is not merely related to the acquisition of basic visuomotor skills during prior practice of the serial reaction time task, but rather to the implicit acquisition of the probabilistic rules that defined stimulus sequences. Moreover, functional connections between the reactivated cuneus and the striatum--the latter being critical for implicit sequence learning--are reinforced during REM sleep after practice on a probabilistic rather than on a random sequence of stimuli. Our results therefore support the hypothesis that REM sleep is deeply involved in the reprocessing and optimization of the high-order information contained in the material to be learned. In addition, we show that the level of acquisition of probabilistic rules attained prior to sleep is correlated to the increase in regional cerebral blood flow during subsequent REM sleep. This suggests that posttraining cerebral reactivation is modulated by the strength of the memory traces developed during the learning episode. Our data provide the first experimental evidence for a link between behavioral performance and cerebral reactivation during REM sleep. PMID- 14527576 TI - Neuroimaging evidence for cortical involvement in the preparation and in the act of swallowing. AB - This study employed whole head magnetoencephalography and synthetic aperture magnetometry to investigate the cortical topography of the preparation and the execution of volitional and reflexive water swallowing and of a simple tongue movement. Concerning movement execution, activation of the mid-lateral primary sensorimotor cortex was strongly lateralized to the left during volitional water swallowing, less strongly lateralized to the left during reflexive water swallowing, and not lateralized at all during tongue movement. In contrast, the preparation for both volitional water swallowing and tongue movement showed a bilateral activation of the primary sensorimotor cortex. No activation was seen prior to reflexive water swallowing. Activation of the left insula and frontal operculum was observed only during both the preparation and the execution of volitional water swallowing. These new findings suggest a left hemispheric dominance for the cortical control of swallowing in humans. PMID- 14527577 TI - Correlates of alpha rhythm in functional magnetic resonance imaging and near infrared spectroscopy. AB - We used simultaneous electroencephalogram-functional magnetic resonance imaging (EEG-fMRI) and EEG-near infrared spectroscopy (NIRS) to investigate whether changes of the posterior EEG alpha rhythm are correlated with changes in local cerebral blood oxygenation. Cross-correlation analysis of slowly fluctuating, spontaneous rhythms in the EEG and the fMRI signal revealed an inverse relationship between alpha activity and the fMRI-blood oxygen level dependent signal in the occipital cortex. The NIRS-EEG measurements demonstrated a positive cross-correlation in occipital cortex between alpha activity and concentration changes of deoxygenated hemoglobin, which peaked at a relative shift of about 8 s. Our data suggest that alpha activity in the occipital cortex is associated with metabolic deactivation. Mapping of spontaneously synchronizing distributed neuronal networks is thus shown to be feasible. PMID- 14527578 TI - Automatic processing of grammar in the human brain as revealed by the mismatch negativity. AB - The Mismatch Negativity (MMN), a neurophysiological indicator of cognitive processing, was used to investigate grammatical processes in the absence of focused attention to language. Subjects instructed to watch a silent video film and to ignore speech stimuli heard grammatical and ungrammatical spoken word strings that were physically identical up to a divergence point where they differed between each other by a minimal acoustic event, the presence or the absence of a final -s sound. The sentence we come was presented as a rare deviant stimulus against the background of frequently occurring ungrammatical strings, and, in a different experiment, the ungrammatical string *we comes was the deviant in the reverse design. To control for effects related to differences between the critical words, come and comes, control conditions were used in which the same words were presented out of linguistic context. At 100-150 ms after the divergence point, the ungrammatical deviant stimulus elicited a larger MMN than the correct sentence at left-anterior recording sites. This difference was not seen under the out-of-context conditions. In the time range 100-400 ms after stimulus divergence, a spatiotemporal pattern of grammatically related effects was documented by statistically significant interactions of the word and context variables. Minimum-Norm Current Estimates of the cortical sources of the grammaticality effects revealed a main source in the left frontal cortex. We use a neurobiological model of serial order processing to provide a tentative explanation for the data. PMID- 14527579 TI - Auditory what, where, and when: a sensory somatotopy in lateral premotor cortex. AB - Recent findings have demonstrated that attention to visual events engages the lateral premotor cortex even in the absence of motor planning. Here, we used functional magnetic resonance imaging to explore acoustically triggered activations within the lateral premotor cortex. Temporal (when), object-related (what), and spatial (where) auditory patterns were to be monitored for violations in a serial prediction task. As a result, we found a modality-dependent modulation for auditory events within the inferior ventrolateral premotor cortex, an area engaged in vocal plans. In addition, however, auditory activations were distributed within the entire premotor cortex depending on which stimulus property was attended to. Attention to where patterns was found to engage fields for gaze and reaching (dorsolateral premotor cortex), what patterns to engage fields for hand movements (superior ventrolateral premotor cortex), and when patterns to engage fields for vocal plans. Together, the findings confirm the idea of a sensory somatotopy in lateral premotor cortex, according to which a perceptual pattern triggers representations within that motor effector which would be most appropriate to generate it as an action effect. PMID- 14527580 TI - A multitrial analysis for revealing significant corticocortical networks in magnetoencephalography and electroencephalography. AB - We present an MEG/EEG framework to reveal statistically significant brain areas engaged in the same cognitive process across trials without resort to averaging procedures. The variability of neuronal responses is assumed to take place only in the reconstructed time series of cortical sources and not in their positions. This hypothesis allows the use of the surrogate data method to detect recurrently active brain areas across trials adjusted with any cortically constrained focal MEEG inverse solution. Results obtained from synthetic data show that considering several trials enhances the accuracy of the source localisation. We apply this approach on MEG data recorded during a simple visual stimulation. The considered stimulus is frequency tagged in order to reveal the neural network correlated to its perception using phase synchronisation analysis. The results show consistent patterns of distributed synchronous networks centred on occipital areas. PMID- 14527581 TI - Bright spots: correlations of gray matter volume with IQ in a normal pediatric population. AB - The localization of brain areas related to cognitive functions has yet to be thoroughly explored in children. We therefore set out to apply volumetric, voxel based, and structural connectivity analyses to magnetic resonance images from a large sample of healthy children. We could confirm a strong correlation of whole brain gray matter volume and the individual intelligence quotient; however, this correlation only developed with age in our sample, in that it was not present in the younger children. With the application of an optimized protocol for voxel based morphometry, the anterior cingulate was shown to be directly correlated with a measure of human intelligence. Furthermore, an analysis of structural connectivity identified gray matter volume in several distinct brain areas to be related to cognitive functions. The implications of our findings for normal development, pathological processes, and our understanding of cognition are discussed and related to previous findings. PMID- 14527582 TI - Preparatory visuo-motor cortical network of the contingent negative variation estimated by current density. AB - The present report studied the intracerebral current density of the contingent negative variation (CNV) during a visuo-manual task using the gap paradigm. The CNV is usually obtained during preparatory periods for perception and action. In this experiment right-hand responses were required. The CNV potential was obtained during the preparatory period from electrodes placed at 58 scalp sites. The CNV showed an early and a late phase. Scalp voltage and source current density maps showed that the early phase was focused on frontal midline sites. The late phase had two foci, one overlying the primary motor cortex and one over occipital sites. When analyzed by low-resolution tomography, the early phase of the CNV showed activations in the supplementary motor area (SMA), the anterior cingulate cortex (ACC), and some posterior areas. The late phase had anterior activations in the left prefrontal cortex, middle frontal cortex, primary motor cortex, ACC, and SMA; and several posterior activations including those in the medial occipital cortex, middle inferior occipital cortex, posterior cingulate cortex, and temporal and parietal areas. Results from the activated areas and their temporal dynamics during the preparatory period suggest that the ACC and the SMA areas recruit the action- and perception-related areas needed to process the expected subsequent imperative task. PMID- 14527583 TI - Human motor cortex activity during mental rotation. AB - The functional role of human premotor and primary motor cortex during mental rotation has been studied using functional MRI at 3 T. Fourteen young, male subjects performed a mental rotation task in which they had to decide whether two visually presented cubes could be identical. Exploratory Fuzzy Cluster Analysis was applied to identify brain regions with stimulus-related time courses. This revealed one dominant cluster which included the parietal cortex, premotor cortex, and dorsolateral prefrontal cortex that showed signal enhancement during the whole stimulus presentation period, reflecting cognitive processing. A second cluster, encompassing the contralateral primary motor cortex, showed activation exclusively after the button press response. This clear separation was possible in 3 subjects only, however. Based on these exploratory results, the hypothesis that primary motor cortex activity was related to button pressing only was tested using a parametric approach via a random-effects group analysis over all 14 subjects in SPM99. The results confirmed that the stimulus response via button pressing causes activation in the primary motor cortex and supplementary motor area while parietal cortex and mesial regions rostral to the supplementary motor area are recruited for the actual mental rotation process. PMID- 14527584 TI - A mathematical approach to the temporal stationarity of background noise in MEG/EEG measurements. AB - The general spatiotemporal covariance matrix of the background noise in MEG/EEG signals is huge. To reduce the dimensionality of this matrix it is modeled as a Kronecker product of a spatial and a temporal covariance matrix. When the number of time samples is larger than, say, J = 500, the iterative Maximum Likelihood estimation of these two matrices is still too time-consuming to be useful on a routine basis. In this study we looked for methods to circumvent this computationally expensive procedure by using a parametric model with subject dependent parameters. Such a model would additionally help with interpreting MEG/EEG signals. For the spatial covariance, models have been derived already and it has been shown that measured MEG/EEG signals can be understood spatially as random processes, generated by random dipoles. The temporal covariance, however, has not been modeled yet, therefore we studied the temporal covariance matrix in several subjects. For all subjects the temporal covariance shows an alpha oscillation and vanishes for large time lag. This gives rise to a temporal noise model consisting of two components: alpha activity and additional random noise. The alpha activity is modeled as randomly occurring waves with random phase and the covariance of the additional noise decreases exponentially with lag. This model requires only six parameters instead of 12 J(J + 1). Theoretically, this model is stationary but in practice the stationarity of the matrix is highly influenced by the baseline correction. It appears that very good agreement between the data and the parametric model can be obtained when the baseline correction window is taken into account properly. This finding implies that the background noise is in principle a stationary process and that nonstationarities are mainly caused by the nature of the preprocessing method. When analyzing events at a fixed sample after the stimulus (e.g., the SEF N20 response) one can take advantage of this nonstationarity by optimizing the baseline window to obtain a low noise variance at this particular sample. PMID- 14527585 TI - Semantic and episodic memory of music are subserved by distinct neural networks. AB - Numerous functional imaging studies have shown that retrieval from semantic and episodic memory is subserved by distinct neural networks. However, these results were essentially obtained with verbal and visuospatial material. The aim of this work was to determine the neural substrates underlying the semantic and episodic components of music using familiar and nonfamiliar melodic tunes. To study musical semantic memory, we designed a task in which the instruction was to judge whether or not the musical extract was felt as "familiar." To study musical episodic memory, we constructed two delayed recognition tasks, one containing only familiar and the other only nonfamiliar items. For each recognition task, half of the extracts (targets) were presented in the prior semantic task. The episodic and semantic tasks were to be contrasted by a comparison to two perceptive control tasks and to one another. Cerebral blood flow was assessed by means of the oxygen-15-labeled water injection method, using high-resolution PET. Distinct patterns of activations were found. First, regarding the episodic memory condition, bilateral activations of the middle and superior frontal gyri and precuneus (more prominent on the right side) were observed. Second, the semantic memory condition disclosed extensive activations in the medial and orbital frontal cortex bilaterally, the left angular gyrus, and predominantly the left anterior part of the middle temporal gyri. The findings from this study are discussed in light of the available neuropsychological data obtained in brain damaged subjects and functional neuroimaging studies. PMID- 14527586 TI - Functional brain imaging of olfactory processing in monkeys. AB - As a step toward bridging the gap between human and animal studies of olfactory brain systems, we report results from an fMRI study of olfaction in squirrel monkeys. High-resolution fMRI images at 3 T with 1.25 x 1.25 x 1.2 mm(3) voxels were obtained covering the whole brain using an 8-cm-diameter birdcage coil and a gradient-echo spiral pulse sequence. Data were acquired from six sedated adult males using a standard block design. All fMRI data were spatially normalized to a common template and analyzed at the individual and group levels with statistical parametric and nonparametric methods. Robust odorant-induced activations were detected in several brain regions previously implicated in conscious human olfactory processing, including the orbitofrontal cortex, cerebellum, and piriform cortex. Consistent with human data, no stimulus intensity effects were observed in any of these regions. Average signal changes in these regions exceeded 0.6%, more than three times the expected signal change based on human fMRI studies of olfaction adjusting for differences in voxel size. These results demonstrate the feasibility of studying olfaction in sedated monkeys with imaging techniques commonly used at 3 T in humans and help promote direct comparisons between humans and nonhuman primates. Our findings, for example, provide novel support for the hypothesis that the cerebellum is involved in sensory acquisition. More broadly, this study suggests that olfactory processing in sedated monkeys and nonsedated humans shares similar neural substrates both within and beyond the primary olfactory system. PMID- 14527587 TI - Spectrotemporal features of the auditory cortex: the activation in response to dynamic ripples. AB - Functional MRI was used to investigate the characteristics of the human cerebral response to dynamic ripples. Dynamic ripples are sound stimuli containing regular spectrotemporal modulations, which are of major importance in speech processing; however, in contrast to speech, dynamic ripples can be characterized fully by a limited number of parameters. Extensive activation consisting of multiple separate regions was found bilaterally in the auditory cortex, particularly along the Heschl's gyri. This agrees with the presence of a structural cortical subdivision into functional fields. The level and the extent of activation were measured and correlated highly (R(2) = 0.97). Both measures depended strongly on the spectral density, temporal frequency, and amplitude of the modulations and matched the perceptual discernibility of the spectrotemporal modulations. The largest responses occurred for parameter values near the optimal human sensitivity. The drift direction of the modulations did not influence the activation. No quantitative differences were found between the two hemispheres. Average brain activation levels proved to be separable with regard to the spectral density and temporal frequency of the modulations. Topographic mappings of the modulation density and frequency onto the cortical surface were shown, approximately in posterolateral-to-anteromedial and lateral-to-medial directions, respectively. Posterolateral regions were most sensitive to spectrotemporal features at a scale similar to phonemes. Anteromedial regions, however, were also relatively sensitive to smaller scale acoustic features. This spatially dependent sensitivity suggests a functional topographic and hierarchical organization of the auditory cortex. PMID- 14527588 TI - Diffusion-weighted magnetic resonance imaging fibre tracking using a front evolution algorithm. AB - A novel technique is presented for estimating white matter connectivity in vivo using diffusion-weighted magnetic resonance imaging. The concept of a fibre orientation density function (ODF) is described, which characterises the uncertainty in the orientation of the underlying white matter fibres, given the set of diffusion-weighted signal intensities at the point of interest. The proposed algorithm is based on the evolution of a front from a seed region, using the information provided by the fibre ODF. Each point reached by the front is assigned an index of connectivity with the seed region. The algorithm was used to track various major white matter fibre pathways in two data sets acquired on the same healthy adult volunteer over separate occasions. Example tracks are shown to illustrate some of the properties of the algorithm, such as robustness to noise and branching capability. Finally, the dependence of the algorithm on the model used to derive the fibre ODF is discussed. PMID- 14527589 TI - Functional reorganization of brain in children affected with congenital hemiplegia: fMRI study. AB - Using functional magnetic resonance imaging, the brain activation related to unilateral sequential finger-to-thumb opposition was studied in six children with a right congenital hemiplegia of cortical origin. They were compared to six age matched controls. In the control group, movements with either hand asymmetrically activated the sensorimotor cortex and premotor areas in both cerebral hemispheres with a typical contralateral predominance. By contrast, paretic finger movements activated both hemispheres in the hemiplegic patients, with a strong ipsilateral predominance favoring the undamaged hemisphere. The activation induced by nonparetic finger movements was restricted to the contralateral undamaged hemisphere. Furthermore, the level of activation in the undamaged cortex was partly related to residual finger dexterity, according to covariance analysis. These activation patterns indicate an adaptive reorganization of the cortical motor networks in this group of patients, with a prominent involvement of the undamaged hemisphere in the control of finger movements with either hand. PMID- 14527590 TI - Brain activity during automatic semantic priming revealed by event-related functional magnetic resonance imaging. AB - Semantic priming occurs when a subject is faster in recognising a target word when it is preceded by a related word compared to an unrelated word. The effect is attributed to automatic or controlled processing mechanisms elicited by short or long interstimulus intervals (ISIs) between primes and targets. We employed event-related functional magnetic resonance imaging (fMRI) to investigate blood oxygen level dependent (BOLD) responses associated with automatic semantic priming using an experimental design identical to that used in standard behavioural priming tasks. Prime-target semantic strength was manipulated by using lexical ambiguity primes (e.g., bank) and target words related to dominant or subordinate meaning of the ambiguity. Subjects made speeded lexical decisions (word/nonword) on dominant related, subordinate related, and unrelated word pairs presented randomly with a short ISI. The major finding was a pattern of reduced activity in middle temporal and inferior prefrontal regions for dominant versus unrelated and subordinate versus unrelated comparisons, respectively. These findings are consistent with both a dual process model of semantic priming and recent repetition priming data that suggest that reductions in BOLD responses represent neural priming associated with automatic semantic activation and implicate the left middle temporal cortex and inferior prefrontal cortex in more automatic aspects of semantic processing. PMID- 14527591 TI - Shannon entropy applied to the analysis of event-related fMRI time series. AB - Event-related functional magnetic resonance imaging (ER-fMRI) refers to the blood oxygen level-dependent (BOLD) signal in response to a short stimulus followed by a long period of rest. These paradigms have become more popular in the last few years due to some advantages over standard block techniques. Most of the analysis of the time series generated in such exams is based on a model of specific hemodynamic response function. In this paper we propose a new method for the analysis of ER-fMRI based in a specific aspect of information theory: the entropy of a signal using the Shannon formulation, which makes no assumption about the shape of the response. The results show the ability to discriminate between activated and resting cerebral regions for motor and visual stimuli. Moreover, the results of simulated data show a more stable pattern of the method, if compared to typical algorithms, when the signal to noise ratio decreases. PMID- 14527592 TI - Effect of head orientation on gaze processing in fusiform gyrus and superior temporal sulcus. AB - We used functional MRI with an event-related design to dissociate the brain activation in the fusiform gyrus (FG) and posterior superior temporal sulcus (STS) for multiple face and gaze orientations. The event-related design allowed for concurrent behavioral analysis, which revealed a significant effect of both head and gaze orientation on the speed of gaze processing, with the face and gaze forward condition showing the fastest reaction times. In conjunction with this behavioral finding, the FG responded with the greatest activation to face and gaze forward, perhaps reflecting the unambiguous social salience of congruent face and gaze directed toward the viewer. Random effects analysis showed greater activation in both the FG and posterior STS when the subjects viewed a direct face compared to an angled face, regardless of gaze direction. Additionally, the FG showed greater activation for forward gaze compared to angled gaze, but only when the face was forward. Together, these findings suggest that head orientation has a significant effect on gaze processing and these effects are manifest not only in the STS, but also the FG. PMID- 14527593 TI - Automated detection of gray matter malformations using optimized voxel-based morphometry: a systematic approach. AB - Malformations of cortical development (MCD) are a recognized cause of epilepsy. Their special significance lies in the fact that, once detected and delineated, they are amenable to surgical removal. However, diagnosis from high-resolution MRI is still difficult, time-consuming, and highly dependent on individual expertise. We have recently proposed a simple procedure to detect cortical dysplasias, using automated procedures available within SPM99 (Wellcome Department, University College London, UK). Here, we aimed to systematically determine the best combination of processing parameters, using an optimized voxel based morphometry approach. We included 20 patients with a known MCD and compared them to a normal database of 53 healthy, age- and gender-matched controls. The approaches taken during spatial normalization and a number of other parameters were systematically altered in order to find the best combination of parameters. Overall, 99 different approaches were evaluated in different ways. As far as possible, automatic processing and evaluation steps were used. With the number of candidate regions for MCD limited to five per patient, the best approaches resulted in the correct identification of up to 16 of 20 malformations. However, a number of approaches failed to perform well. The reasons for these failures and the implications this has for other studies are discussed. We conclude that voxel based morphometry is able to detect cortical malformations with a high degree of accuracy. However, specific problems seem to arise when using an optimized protocol for voxel-based morphometry, indicating that this protocol may not be optimal for all voxel-based studies on brain morphology. Our approach, involving systematic alterations of parameters and evaluation, may be useful for other studies. PMID- 14527594 TI - A right hemispheric frontocerebellar network for time discrimination of several hundreds of milliseconds. AB - Debate still surrounds the nature of the role of the dorsolateral prefrontal gyrus (DLPFC) in time perception. This region is frequently associated with working memory and is thus implicated as a so-called "accumulator" within a hypothesized internal clock model. However, we hypothesized that this region may have a more primary role in time perception. To test this hypothesis we used functional magnetic resonance imaging (fMRI) to examine the neural correlates of relatively pure time perception with a temporal discrimination task where intervals of 1 s had to be discriminated from those of 1.3, 1.4, and 1.5 s. Time perception in this particular time domain within the "perceived present" has not previously been investigated using fMRI. By using relatively short time periods to be discriminated and also contrasting activation with an order judgment task, we aimed to minimize the confounding aspects of sustained attention and working memory. In a group of 20 healthy right-handed adult males, neural activation associated with time discrimination was found in a predominantly right hemispheric network of right dorsolateral and inferior prefrontal cortices, right supplementary motor area, and left cerebellum. We conclude that right DLPFC, rather than having a purely working memory function, might be more centrally involved in time perception than previously thought. PMID- 14527595 TI - Right inferior prefrontal cortex mediates response inhibition while mesial prefrontal cortex is responsible for error detection. AB - Inhibitory control and error detection are among the highest evolved human self monitoring functions. Attempts in functional neuroimaging to effectively isolate inhibitory motor control from other cognitive functions have met with limited success. Different brain regions in inferior, mesial, and dorsolateral prefrontal cortices and parietal and temporal lobes have been related to inhibitory control in go/no-go and stop tasks. The widespread activation reflects the fact that the designs used so far have comeasured additional noninhibitory cognitive functions such as selective attention, response competition, decision making, target detection, and inhibition failure. Here we use rapid, mixed trial, event-related functional magnetic resonance imaging to correlate brain activation with an extremely difficult situation of inhibitory control in a challenging stop task that controls for noninhibitory functions. The difficulty of the stop task, requiring withholding of a triggered motor response, was assured by an algorithm that adjusted the task individually so that each subject only succeeded on half of all stop trials, failing on the other half. This design allowed to elegantly separate brain activation related to successful motor response inhibition and to inhibition failure or error detection. Brain activation correlating with successful inhibitory control in 20 healthy volunteers could be isolated in right inferior prefrontal cortex. Failure to inhibit was associated with activation in mesial frontopolar and bilateral inferior parietal cortices, presumably reflecting an attention network for error detection. PMID- 14527596 TI - Dynamic programming generation of boundaries of local coordinatized submanifolds in the neocortex: application to the planum temporale. AB - Dynamic programming is used to define boundaries of cortical submanifolds with focus on the planum temporale (PT) of the superior temporal gyrus (STG), which has been implicated in a variety of neuropsychiatric disorders. To this end, automated methods are used to generate the PT manifold from 10 high-resolution MRI subvolumes ROI masks encompassing the STG. A procedure to define the subvolume ROI masks from original MRI brain scans is developed. Bayesian segmentation is then used to segment the subvolumes into cerebrospinal fluid, gray matter (GM), and white matter (WM). 3D isocontouring using the intensity value at which there is equal probability of GM and WM is used to reconstruct the triangulated graph representing the STG cortical surface, enabling principal curvature at each point on the graph to be computed. Dynamic programming is used to delineate the PT manifold by tracking principal curves from the retro-insular end of the Heschl's gyrus (HG) to the STG, along the posterior STG up to the start of the ramus and back to the retro-insular end of the HG. A coordinate system is then defined on the PT manifold. The origin is defined by the retro insular end of the HG and the y-axis passes through the point on the posterior STG where the ramus begins. Automated labeling of GM in the STG is robust with L(1) distances between Bayesian and manual segmentation in the range 0.001-0.12 (n = 20). PT reconstruction is also robust with 90% of the vertices of the reconstructed PT within about 1 voxel (n = 20) from semiautomated contours. Finally, the reliability index (based on interrater intraclass correlation) for the surface area derived from repeated reconstructions is 0.96 for the left PT and 0.94 for the right PT, thus demonstrating the robustness of dynamic programming in defining a coordinate system on the PT. It provides a method with potential significance in the study of neuropsychiatric disorders. PMID- 14527597 TI - FMRI investigation of cross-cultural music comprehension. AB - The popular view of music as a "universal" language ignores the privileged position of the cultural insider in comprehending musical information unique to their own tradition. The purpose of this study was to test the hypothesis that listeners would demonstrate different neural activity in response to culturally familiar and unfamiliar music and that those differences may be affected by the extent of subjects' formal musical training. Just as familiar languages have been shown to use distinct brain processes, we hypothesized that an analogous difference might be found in music and that it may depend in part on subjects' formal musical knowledge. Using fMRI we compared the activation patterns of professional musicians and untrained controls reared in the United States as they listened to music from their culture (Western) and from an unfamiliar culture (Chinese). No overall differences in activation were observed for either subject group in response to the two musical styles, although there were differences in recall performance based on style and there were activation differences based on training. Trained listeners demonstrated additional activation in the right STG for both musics and in the right and left midfrontal regions for Western music and Chinese music, respectively. Our findings indicate that listening to culturally different musics may activate similar neural resources but with dissimilar results in recall performance. PMID- 14527598 TI - A voxel-based morphometric MRI study in female patients with borderline personality disorder. AB - Subtle prefrontal and limbic structural abnormalities have been reported in borderline personality disorder (BPD). In order to further validate the previously reported findings and to more precisely describe the nature of the structural change we performed a voxel-based morphometric (VBM) study in patients with BPD. Twenty female patients with BPD and 21 female healthy controls were investigated. High-resolution 3-D datasets were acquired and analyzed following an optimized protocol of VBM in the framework of statistical parametric mapping (SPM99). Gray matter volume loss was found in the left amygdala. No other differences in gray or white matter volume or density were found anywhere else in the brain. Our findings support the hypothesis that temporolimbic abnormalities play a role in the pathophysiology of BPD. Prefrontal structural alterations in BPD were not observed in this study. PMID- 14527599 TI - A voxel-based morphometric study of nondemented adults with Down Syndrome. AB - Previous structural brain imaging studies of Down Syndrome (DS) have offered important insights into the underlying morphometric aberrations associated with the condition. These previous studies have relied almost exclusively on classic region-of-interest (ROI)-based morphometry, a method in which a finite number of anatomical structures must be defined and delineated a priori. Here we use the fully automated voxel-based morphometry (VBM) approach on 19 nondemented individuals with DS and 11 age-matched controls in order to provide a full-brain assessment of DS morphology. Foci of statistically significant (P < 0.05, corrected for multiple comparisons) reductions in gray matter (GM) tissue were observed in the cerebellum, cingulate gyrus, left medial frontal lobe, right middle/superior temporal gyrus, and the left CA2/CA3 region of the hippocampus. Significant decreases in white matter (WM) tissue were noted throughout the inferior brainstem. Foci of statistically significant (P < 0.05, corrected for multiple comparisons) increases in GM tissue were observed in a superior/caudal portion of the brainstem and left parahippocampal gyrus. Significant increases in WM tissue were noted bilaterally in the parahippocampal gyrus. We also noted significant increases in cerebral spinal fluid in regions suggesting enlarged lateral ventricles in the DS group. While these results are generally consistent with prior ROI-based imaging studies of nondemented DS individuals, the present findings provide additional understanding of the three-dimensional topography of DS morphology throughout the brain. The consistency of these findings with prior imaging reports demonstrates the utility of the VBM technique for investigating the neuroanatomy of DS. PMID- 14527600 TI - The preparation and readiness for voluntary movement: a high-field event-related fMRI study of the Bereitschafts-BOLD response. AB - Activity within motor areas of the cortex begins to increase 1 to 2 s prior to voluntary self-initiated movement (termed the Bereitschaftspotential or readiness potential). There has been much speculation and debate over the precise source of this early premovement activity as it is important for understanding the roles of higher order motor areas in the preparation and readiness for voluntary movement. In this study, we use high-field (3-T) event-related fMRI with high temporal sampling (partial brain volumes every 250 ms) to specifically examine hemodynamic response time courses during the preparation, readiness, and execution of purely self-initiated voluntary movement. Five right-handed healthy volunteers performed a rapid sequential finger-to-thumb movement performed at self-determined times (12-15 trials). Functional images for each trial were temporally aligned and the averaged time series for each subject was iteratively correlated with a canonical hemodynamic response function progressively shifted in time. This analysis method identified areas of activation without constraining hemodynamic response timing. All subjects showed activation within frontal mesial areas, including supplementary motor area (SMA) and cingulate motor areas, as well as activation in left primary sensorimotor areas. The time courses of hemodynamic responses showed a great deal of variability in shape and timing between subjects; however, four subjects clearly showed earlier relative hemodynamic responses within SMA/cingulate motor areas compared with left primary motor areas. These results provide further evidence that the SMA and cingulate motor areas are major contributors to early stage premovement activity and play an important role in the preparation and readiness for voluntary movement. PMID- 14527601 TI - High b value diffusion-weighted imaging is more sensitive to white matter degeneration in Alzheimer's disease. AB - It has been reported that diffusion-weighted imaging (DWI) can detect white matter degeneration in the Alzheimer's disease (AD) brain. We hypothesized that imaging of the slow diffusion component using high b value DWI is more sensitive to AD-related white matter degeneration than is conventional DWI, and therefore we studied the effects of high b value on lesion-to-normal contrast and contrast to-noise ratio (CNR). Seven AD patients and seven age-matched normal subjects were studied with full-tensor DWI at three different b values (1000, 2000, and 4000 s/mm(2)) without changing echo time or diffusion time, and the mean diffusivities in the parietal and occipital regions were measured. Statistical analyses revealed that use of higher b values significantly improves both lesion to-normal contrast and CNR. We concluded that high b value DWI is more sensitive to AD-related white matter degeneration than is conventional DWI. PMID- 14527602 TI - Adolescent immaturity in attention-related brain engagement to emotional facial expressions. AB - Selective attention, particularly during the processing of emotionally evocative events, is a crucial component of adolescent development. We used functional magnetic resonance imagining (fMRI) with adolescents and adults to examine developmental differences in activation in a paradigm that involved selective attention during the viewing of emotionally engaging face stimuli. We evaluated developmental differences in neural activation for three comparisons: (1) directing attention to subjective responses to fearful facial expressions relative to directing attention to a nonemotional aspect (nose width) of fearful faces, (2) viewing fearful relative to neutral faces while attending to a nonemotional aspect of the face, and (3) viewing fearful relative to neutral faces while attention was unconstrained (passive viewing). The comparison of activation across attention tasks revealed greater activation in the orbital frontal cortex in adults than in adolescents. Conversely, when subjects attended to a nonemotional feature, fearful relative to neutral faces influenced activation in the anterior cingulate more in adolescents than in adults. When attention was unconstrained, adolescents relative to adults showed greater activation in the anterior cingulate, bilateral orbitofrontal cortex, and right amygdala in response to the fearful relative to neutral faces. These findings suggest that adults show greater modulation of activity in relevant brain structures based on attentional demands, whereas adolescents exhibit greater modulation based on emotional content. PMID- 14527603 TI - Temporal integration of sequential auditory events: silent period in sound pattern activates human planum temporale. AB - Temporal integration is a fundamental process that the brain carries out to construct coherent percepts from serial sensory events. This process critically depends on the formation of memory traces reconciling past with present events and is particularly important in the auditory domain where sensory information is received both serially and in parallel. It has been suggested that buffers for transient auditory memory traces reside in the auditory cortex. However, previous studies investigating "echoic memory" did not distinguish between brain response to novel auditory stimulus characteristics on the level of basic sound processing and a higher level involving matching of present with stored information. Here we used functional magnetic resonance imaging in combination with a regular pattern of sounds repeated every 100 ms and deviant interspersed stimuli of 100-ms duration, which were either brief presentations of louder sounds or brief periods of silence, to probe the formation of auditory memory traces. To avoid interaction with scanner noise, the auditory stimulation sequence was implemented into the image acquisition scheme. Compared to increased loudness events, silent periods produced specific neural activation in the right planum temporale and temporoparietal junction. Our findings suggest that this area posterior to the auditory cortex plays a critical role in integrating sequential auditory events and is involved in the formation of short-term auditory memory traces. This function of the planum temporale appears to be fundamental in the segregation of simultaneous sound sources. PMID- 14527604 TI - Predominant ventromedial frontopolar metabolic impairment in frontotemporal dementia. AB - In a multicenter study, FDG-PET images in a population of 29 patients with frontotemporal dementia (FTD) were compared to controls with similar age from each center. A conjunction analysis led to identification of the ventromedial frontopolar cortex as the single region affected in each and every FTD patients. This precise regional metabolic impairment should be integrated with recent neuropsychological researches, such as those showing that the ventromedial frontal cortex is critically involved in decision-making processes based on personal experience, feelings of rightness or social knowledge, processes that are characteristically impaired in FTD. PMID- 14527605 TI - Left-hemisphere dominance in early nociceptive processing in the human parasylvian cortex. AB - Pain perception comprises sensory and emotional dimensions. While the emotional experience is thought to be represented in the right hemisphere, we here report a left-hemisphere dominance for the early sensory component of pain perception using brain electrical source analysis of laser-evoked potentials. Ten right handed subjects underwent several series of laser radiant heat stimuli to pairs of parallel lines on the dorsum of the left or right hand. Stimulus location and intensity were randomised independently. The sensory-discriminative aspects of pain were emphasised by asking the subjects to perform either a spatial or an intensity discrimination task and were contrasted with active distraction by mental arithmetics. Pain ratings obtained after each of the laser stimulus series revealed an analgesic effect of distraction (27%, P < 0.001). Four equivalent dipole sources were active in the latency range of 100-200 ms (bilateral operculoinsular cortex, midcingulate gyrus, postcentral gyrus). The sources in the operculoinsular cortex exhibited (a) the shortest peak latency (155 +/- 6 ms), (b) the most pronounced enhancement during spatial and intensity discrimination tasks compared to active distraction (43%, P < 0.001), and (c) a significantly stronger source activity in the left hemisphere independent of stimulation side (23%, P < 0.05). The distribution of these sources extended into the dorsal insula. The postcentral source had the longest peak latency (180 +/- 7 ms); its source strength was task-dependent (25%, P = 0.051) but exhibited no hemisphere dominance. The midcingulate source had an intermediate peak latency (169 +/- 7 ms). Its source strength was modulated by tasks, but this modulation was significant only in the latency range >200 ms (46%, P < 0.001). These findings suggest a dominant role of the left frontal operculum and adjacent dorsal insula in the early sensory-discriminative dimensions of pain processing. This region has been proposed to be the cortical projection target of nociceptive pathways from the spinal cord to the ventroposteroinferior and ventromedial (its posterior part: VMpo) thalamic nuclei. PMID- 14527606 TI - Region growing method for the analysis of functional MRI data. AB - Existing analytical techniques for functional magnetic resonance imaging (fMRI) data always need some specific assumptions on the time series. In this article, we present a new approach for fMRI activation detection, which can be implemented without any assumptions on the time series. Our method is based on a region growing method, which is very popular for image segmentation. A comparison of performance on fMRI activation detection is made between the proposed method and the deconvolution method and the fuzzy clustering method with receiver operating characteristic (ROC) methodology. In addition, we examine the effectiveness and usefulness of our method on real experimental data. Experimental results show that our method outperforms over the deconvolution method and the fuzzy clustering method on a number of aspects. These results suggest that our region growing method can serve as a reliable analysis of fMRI data. PMID- 14527607 TI - A disturbance of nonlinear interdependence in scalp EEG of subjects with first episode schizophrenia. AB - It has been proposed that schizophrenia arises through a disturbance of coupling between large-scale cortical systems. This "disconnection hypothesis" is tested by applying a measure of dynamical interdependence to scalp EEG data. EEG data were collected from 40 subjects with a first episode of schizophrenia and 40 matched healthy controls. An algorithm for the detection of dynamical interdependence was applied to six pairs of bipolar electrodes in each subject. The topographic organization of the interdependence was calculated and served as the principle measure of cortical integration. The rate of occurrence of dynamical interdependence did not statistically differ between subject groups at any of the sites. However, the topography across the scalp was significantly different between the two groups. Specifically, nonlinear interdependence tended to occur in larger concurrent "clusters" across the scalp in schizophrenia than in the healthy subjects. This disturbance was reflected most strongly in left intrahemispheric coupling and did not differ significantly according to symptomatology. Medication dose and subject arousal were not observed to be confounding factors. The study of dynamical interdependence in scalp EEG data does not support a straightforward interpretation of the disconnection hypothesis that there is a decrease in the strength of functional coupling between adjacent cortical regions. Rather, it suggests a dysregulation in the organization of dynamical interactions across supraregional brain systems. PMID- 14527608 TI - Differences in the hemodynamic response to event-related motor and visual paradigms as measured by near-infrared spectroscopy. AB - Several current brain imaging techniques rest on the assumption of a tight coupling between neural activity and hemodynamic response. The nature of this neurovascular coupling, however, is not completely understood. There is some evidence for a decoupling of these processes at the onset of neural activity, which manifests itself as a momentary increase in the relative concentration of deoxyhemoglobin (HbR). The existence of this early component of the hemodynamic response function, however, is controversial, as it is inconsistently found. Near infrared spectroscopy (NIRS) allows quantification of levels of oxyhemoglobin (HbO(2)) and HbR during task performance in humans. We acquired NIRS data during performance of simple motor and visual tasks, using rapid-presentation event related paradigms. Our results demonstrate that rapid, event-related NIRS can provide robust estimates of the hemodynamic response without artifacts due to low frequency signal components, unlike data from blocked designs. In both the motor and visual data the onset of the increase in HbO(2) occurs before HbR decreases, and there is a poststimulus undershoot. Our results also show that total blood volume (HbT) drops before HbO(2) and undershoots baseline, raising a new issue for neurovascular models. We did not find early deoxygenation in the motor data using physiologically plausible values for the differential pathlength factor, but did find one in the visual data. We suggest that this difference, which is consistent with functional magnetic resonance imaging (fMRI) data, may be attributable to different capillary transit times in these cortices. PMID- 14527609 TI - Improving tissue segmentation of human brain MRI through preprocessing by the Gegenbauer reconstruction method. AB - The Gegenbauer image reconstruction method, previously shown to improve the quality of magnetic resonance images, is utilized in this study as a segmentation preprocessing step. It is demonstrated that, for all simulated and real magnetic resonance images used in this study, the Gegenbauer reconstruction method improves the accuracy of segmentation. Although it is more desirable to use the k space data for the Gegenbauer reconstruction method, only information acquired from MR images is necessary for the reconstruction, making the procedure completely self-contained and viable for all human brain segmentation algorithms. PMID- 14527610 TI - Activation of the human posterior parietal and temporoparietal cortices during audiotactile interaction. AB - We recorded cortical-evoked responses with a whole-scalp neuromagnetometer to study human brain dynamics associated with audiotactile interaction. The subjects received unilateral auditory (A) or tactile (T) stimuli, or both stimuli simultaneously (AT), alternating to the left and right side. Responses to AT stimuli differed significantly from the algebraic sum of responses to A and T stimuli (A + T) at 75-85 and 105-130 ms and indicated suppressive audiotactile interaction. Source modeling revealed that the earlier interaction occurred in the contralateral posterior parietal cortex and the later interaction in the contralateral parietal opercula between the SII cortex and the auditory cortex. The interaction was significantly stronger in the left than the right hemisphere. In most subjects, AT responses were far more similar to T than to A responses, suggesting suppression of auditory processing during the spatially and temporally concordant audiotactile stimuli in which the tactile component was subjectively more salient. PMID- 14527611 TI - Characterization of sexual dimorphism in the human corpus callosum. AB - Despite decades of research, there is still no agreement over the presence of gender-based morphologic differences in the human corpus callosum. We approached the problem using a highly precise computational technique for shape comparison. Starting with a prospectively acquired sample of cranial MRIs of healthy volunteers (age ranges 18-84), the variations of individual callosa are quantified with respect to a reference callosum shape in the form of Jacobian determinant maps derived from the geometric transformations that map the reference callosum into anatomic alignment with the subject callosa. Voxelwise t tests performed over the determinant values demonstrated that females had a larger splenium than males (P < 0.001 uncorrected for multiple comparisons) while males possessed a larger genu (P < 0.001). In addition, pointwise Pearson plots using age as a correlate showed a different pattern of age-related changes in male and female callosa, with female splenia tending to expand more with age, while the male genu tended to contract. Our results demonstrate significant morphologic differences in the corpus callosum between genders and a possible sex difference in the neuro-developmental cycle. PMID- 14527612 TI - The functional magnetic resonance imaging hemodynamic response to faces remains stable until the ninth decade. AB - The effects of aging on blood oxygen level dependent signal changes and the hemodynamic response (HDR) remain controversial. Using functional magnetic resonance (MR) imaging, we examined the HDR properties and activated voxel counts in striate and extrastriate cortex in 18 healthy elderly subjects in response to a simple visual paradigm. Subjects of equal number and gender were prospectively separated into groups from the seventh, eighth, and ninth decades. Activation data were compared with those of 6 healthy subjects aged 30-39 under the same conditions. We found no systematic difference in HDR amplitude, shape, or latency across these groups. However, increasing age over 60 was associated with a significant decline in activated voxel counts, relative to the young controls. The results are discussed in comparison with previously published studies and in the context of the effects of aging on MR signal change. While robust activation can be produced in the striate and extrastriate cortices until the end of the ninth decade, caution should be exercised when comparing data from subjects in different decades. As functional magnetic resonance imaging is increasingly being used to examine patients with stroke and dementia, these results emphasize the importance of careful selection and age matching of control subjects when comparing with a patient population affected by disease processes associated with aging. PMID- 14527613 TI - Structural (operational) synchrony of EEG alpha activity during an auditory memory task. AB - Memory paradigms are often used in psycho-physiological experiments in order to understand the neural basis underlying cognitive processes. One of the fundamental problems encountered in memory research is how specific and complementary cortical structures interact with each other during episodic encoding and retrieval. A key aspect of the research described below was estimating the coupling of rapid transition processes (in terms of EEG description) which occur in separate cortical areas rather than estimating the routine phase-frequency synchrony in terms of correlation and coherency. It is assumed that these rapid transition processes in the EEG amplitude correspond to the "switching on/off" of brain elemental operations. By making a quantitative estimate of the EEG structural synchrony of alpha-band power between different EEG channels, it was shown that short-term memory has the emergent property of a multiregional neuronal network, and is not the product of strictly hierarchical processing based on convergence through association regions. Moreover, it was demonstrated that the dynamic temporal structure of alpha activity is strongly correlated to the dynamic structure of working memory. PMID- 14527614 TI - The effect of acetazolamide on the changes of cerebral blood flow and oxygen metabolism during visual stimulation. AB - Acetazolamide, a carbonic anhydrase inhibitor, has an anticonvulsant effect which may result from a decrease in the efficacy of synaptic transmission due to a decrease of pH. Our previous study showed that acetazolamide induced a significant increase in global and regional cerebral blood flow (CBF), but caused no significant change in the cerebral metabolic rate of oxygen (CMRO(2)). To investigate the effect of acetazolamide on the responses of CBF and CMRO(2) during neural stimulation, we used positron emission tomography to measure CBF and CMRO(2) in six normal volunteers at the fixation-only baseline visual state and during visual stimulation before and after administration of 1 g of acetazolamide. Visual stimulation induced a significant increase in CBF (33%) in the visual cortex compared with baseline values, but caused no significant change in CMRO(2), while no significant change in global CBF or CMRO(2) was found. During visual stimulation after acetazolamide administration, both global and visual cortical CBF and CMRO(2) showed similar changes compared with the respective baseline values (37 and 65% increases in CBF and 8 and 16% decreases in CMRO(2), respectively). When corrected by the global values, the magnitudes of the CBF and CMRO(2) responses to visual stimulation after acetazolamide administration were less than those before (20% vs 38% in CBF and -9% vs 3% in CMRO(2)). Considering our previous observation that the effect of acetazolamide was similar throughout cerebral cortical regions, we suggest that acetazolamide decreases the responses of both CBF and CMRO(2) during visual stimulation, which indicates that this drug may affect neuronal excitability. PMID- 14527615 TI - Repeated premotor rTMS leads to cumulative plastic changes of motor cortex excitability in humans. AB - We explored the aftereffects of two premotor 1 Hz rTMS sessions on motor cortex excitability in healthy humans. In experiment 1, 11 healthy right-handed volunteers received 20-min submotor threshold 1 Hz rTMS trains over the left premotor cortex on 2 consecutive days. Left motor cortex excitability was determined at baseline, immediately after, 30, 60, 120 min, and 24 h after each rTMS session. We measured motor thresholds, amplitudes of motor-evoked potentials, silent periods, and paired-pulse excitability at interstimulus intervals (ISI) of 3-7, 10, and 15 ms. In experiment 2, 5 volunteers received two identical rTMS trains on Days 1 and 7. Measurements were carried out on Day 1 (first rTMS train), Day 2, and Day 7 (second rTMS train). In experiment 1 there was a selective increase of paired pulse facilitation at an ISI of 7 ms after rTMS lasting for less than 30 min on Day 1. This effect was also present after rTMS on Day 2. However, it persisted for at least 2 h. In experiment 2 the same extra facilitation was induced by rTMS on Days 1 and 7 but not on Day 2. It lasted for less than 30 min on both Day 1 and Day 7. We conclude that 1 Hz premotor rTMS leads to cumulative plastic changes of intrinsic motor cortex excitability when repeated within 24 h but not after 1 week, implying the formation of memory after the first rTMS train lasting more than a day but less than a week. PMID- 14527616 TI - Dynamics of cognitive processing in the human hippocampus by neuromagnetic and neurochemical assessments. AB - Various features of cognitive processing have been studied using event-related electric potentials and magnetoencephalography (MEG), as well as neuroimaging. Magnetic resonance spectroscopy (MRS) is a unique, noninvasive approach of measuring cellular metabolism that reflects the static metabolic state. The present study is the first to show noninvasively the dynamic neurophysiological and metabolic changes that occur during cognitive processing in vivo in the human hippocampus, as measured by MEG and spin-echo dynamic (1)H-MRS time-locked to the onset of the stimulus. The stimuli consisted of unpleasant and pleasant pictures of faces of human or primate babies. The event-related synchronization of theta activity and levels of creatine/phosphocreatine and choline-containing compounds relative to the respective level in the resting condition increased significantly, more in the right hippocampus than in the left, during the target discrimination task and also more in the right hippocampus in response to the unpleasant target picture than the pleasant one. These results suggest that excitatory postsynaptic metabolism in the hippocampus, especially in the right hippocampus, is involved in discriminative and cognitive processing of emotional information. This newly devised method combining event-related MEG with MRS can be used to noninvasively elucidate the dynamic features of neurophysiology and neurochemical metabolism and represents a promising approach toward improving our understanding of brain pathophysiology. PMID- 14527617 TI - Equivalence of cognitive processes in brain imaging and behavioral studies: evidence from task switching. AB - A growing number of studies on the higher-order cognitive functions of the human brain use brain-imaging techniques, such as functional magnetic resonance imaging (fMRI). For the validity and generality of fMRI results, it is important that the relevant cognitive processes are equivalent to those functioning in typical settings used in behavioral research. This equivalence could be, for example, endangered by different spatial frames of reference when lying in the scanner. In the present study, we tested whether the cognitive processes, as reflected in behavioral data in brain-imaging settings, are indeed functionally equivalent to those reflected in "purely" behavioral settings. To this end, we used a task switching paradigm with a spatial component, increasing the likelihood to find effects of experimental setting. We compared the data of three different groups that only differed in testing environments (real, operating fMRI vs simulated fMRI vs standard behavioral with upright position of participants) but used otherwise strictly equivalent experimental conditions. Of importance for our validation purposes, unlike previous studies, we included a group with a behavioral setting, and we tested whether we would replicate a nontrivial, complex three-way interaction across all three groups. We replicated the predicted complex data pattern in all groups, suggesting functional equivalence of the underlying cognitive processes. We also found strongly increased reaction time (RT) levels in the two fMRI groups. We attribute this increase to unspecific distracting factors affecting late motor processes and discuss potential methodological implications of this increased baseline RT in the scanner. PMID- 14527618 TI - Selective enhancement of functional connectivity in the left prefrontal cortex during sentence processing. AB - We present the results of correlation analyses for identifying temporally correlated activations between multiple regions of interest. We focused on functional connectivity for two regions in the prefrontal cortex: the left inferior frontal gyrus (L. F3t/F3O) and the left precentral sulcus (L. PrCS). Temporal correlations of functional magnetic resonance imaging signals were separately examined during a sentence comprehension task and a lexical decision task, thereby averaging data throughout all voxels within a region of interest used as a reference region. We found that the reciprocal connectivity between L. F3t/F3O and L. PrCS was significantly enhanced during sentence processing, but not during lexico-semantic processing, which was confirmed under both auditory and visual conditions. Furthermore, significantly correlated regions were mostly concentrated in the left prefrontal cortex during the sentence task. These results demonstrate that the functional connectivity within the left prefrontal cortex is selectively enhanced for processing sentences, which may subserve the use of syntactic information for integrating lexico-semantic information. PMID- 14527619 TI - Minipig negative slow wave demonstrates target/nontarget differences in P300 paradigm. AB - The negative slow wave (NSW) is a late component of the event-related potential (ERP) in man modulated like the P300 by the stimulus, the task, and the response demand. Aiming at the development of a minipig model of schizophrenia, we investigated scalp ERPs in an auditory P300 paradigm in six Gottingen minipigs. Before training, we observed no difference between target and nontarget NSW. After training, target NSW amplitude was increased 50% compared to nontarget. A P350 was recognized, but the finding of a lack of target/nontarget difference is not conclusive. PMID- 14527620 TI - A heuristic for the degrees of freedom of statistics based on multiple variance parameters. AB - In neuroimaging, data are often modeled using general linear models. Here, we focus on GLMs with error covariances which are modeled as a linear combination of multiple variance/covariance components. Each of these components is weighted by one variance parameter. In many analyses variance parameters are estimated using restricted maximum likelihood (ReML). Most classical approaches assume the error covariance matrix can be factorized into a single variance parameter and a nonspherical correlation matrix. In this context, the F test based on a single variance parameter, with a suitable correction to the degrees of freedom, is the standard inference tool. This correction can also be adapted to models with multiple variance parameters. However, this extension overlooks the uncertainty about the variance parameter estimates and P values tend to be underestimated. Here, we show how one can overcome this problem to render the F test more exact. This issue is important, because serial correlations in fMRI time series are generally modeled using multiple variance parameters. Another application is to hierarchical linear models, which are used for modeling multisubject data. To illustrate our approach, we apply it to some typical modeling scenarios in fMRI data analysis. PMID- 14527621 TI - Inferior central sulcus: variations of anatomy and function on the example of the motor tongue area. AB - We wanted to define the position of the primary motor tongue area (MTA) by using functional magnetic resonance imaging (fMRI) to display the MTA in relation to the inferolateral segment of the central sulcus (CS). The anatomy of the inferolateral segment was analyzed in 24 healthy subjects, using the magnetization prepared rapid acquisition gradient echo sequence. The position of the MTA was defined in 11 subjects by using fMRI to identify the sites of maximal activation for each subject in relation to that subject's own CS. The MTA was then displayed in three orthogonal planes, and in lateral surface reformations. The inferolateral segment displayed two distinct curves in 33 of 48 (69%) hemispheres, three curves in 6%, and four curves in 23%. Significant paradigm correlated activations were found in every hemisphere. Thereof 89 local maxima were determined, with 84 (94%) located in the region of the CS. Sixty-seven (80%) of the 84 CS activations lay along the two lowest curves of the CS. In 86% of cases, activations situated in the middle and deep portion of the anterior bank of the CS were encompassed within the three axial sections centered on the cella media of the lateral ventricles. In conclusion, the variability of the inferior CS segment precluded the assignment of the MTA to a specific anatomic configuration. However, the position of the MTA could be approximated by the intersection between the CS and the three axial planes through, just above, and just below the cella media of the lateral ventricles. PMID- 14527622 TI - Nitric oxide-an endogenous contrast agent contributing to "the elusive initial dip?". PMID- 14527624 TI - Origin and diversity of human physiological adaptability. PMID- 14527625 TI - Metabolic correlates of hominid brain evolution. AB - Large brain sizes in humans have important metabolic consequences as humans expend a relatively larger proportion of their resting energy budget on brain metabolism than other primates or non-primate mammals. The high costs of large human brains are supported, in part, by diets that are relatively rich in energy and other nutrients. Among living primates, the relative proportion of metabolic energy allocated to the brain is positively correlated with dietary quality. Humans fall at the positive end of this relationship, having both a very high quality diet and a large brain size. Greater encephalization also appears to have consequences for aspects of body composition. Comparative primate data indicate that humans are 'under-muscled', having relatively lower levels of skeletal muscle than other primate species of similar size. Conversely, levels of body fatness are relatively high in humans, particularly in infancy. These greater levels of body fatness and reduced levels of muscle mass allow human infants to accommodate the growth of their large brains in two important ways: (1) by having a ready supply of stored energy to 'feed the brain', when intake is limited and (2) by reducing the total energy costs of the rest of the body. Paleontological evidence indicates that the rapid brain evolution observed with the emergence of Homo erectus at approximately 1.8 million years ago was likely associated with important changes in diet and body composition. PMID- 14527626 TI - Survival of the fattest: fat babies were the key to evolution of the large human brain. AB - In the past 2 million years, the hominid lineage leading to modern humans evolved significantly larger and more sophisticated brains than other primates. We propose that the modern human brain was a product of having first evolved fat babies. Hence, the fattest (infants) became, mentally, the fittest adults. Human babies have brains and body fat each contributing to 11-14% of body weight, a situation which appears to be unique amongst terrestrial animals. Body fat in human babies provides three forms of insurance for brain development that are not available to other land-based species: (1) a large fuel store in the form of fatty acids in triglycerides; (2) the fatty acid precursors to ketone bodies which are key substrates for brain lipid synthesis; and (3) a store of long chain polyunsaturated fatty acids, particularly docosahexaenoic acid, needed for normal brain development. The triple combination of high fuel demands, inability to import cholesterol or saturated fatty acids, and dependence on docosahexaenoic acid puts the mammalian brain in a uniquely difficult situation compared with other organs and makes its expansion in early humans all the more remarkable. We believe that fresh- and salt-water shorelines provided a uniquely rich, abundant and accessible food supply, and the only viable environment for evolving both body fat and larger brains in human infants. PMID- 14527627 TI - Differential resource utilization by extant great apes and australopithecines: towards solving the C4 conundrum. AB - Morphological and biogeochemical evidence suggest that australopithecines had diets markedly different from those of extant great apes. Stable carbon isotope analysis, for example, has shown that significant amounts of the carbon consumed by australopithecines were derived from C(4) photosynthesis in plants. This means that australopithecines were eating large quantities of C(4) plants such as tropical grasses and sedges, or were eating animals that were themselves eating C(4) plants. In contrast, there is no evidence that modern apes consume appreciable amounts of any of these foods, even in the most arid extents of their ranges where these foods are most prevalent. Environmental reconstructions of early australopithecine environments overlap with modern chimpanzee habitats. This, in conjunction with the stable isotope evidence, suggests that australopithecines and great apes, even in similar environments, would utilize available resources differently. Thus, the desire or capacity to use C(4) foods may be a basal character of our lineage. We do not know, however, which of the nutritionally disparate C(4) foods were utilized by hominids. Here we discuss which C(4) resources were most likely consumed by australopithecines, as well as the potential nutritional, physiological, and social consequences of eating these foods. PMID- 14527628 TI - 'Cooking as a biological trait'. AB - No human foragers have been recorded as living without cooking, and people who choose a 'raw-foodist' life-style experience low energy and impaired reproductive function. This suggests that cooking may be obligatory for humans. The possibility that cooking is obligatory is supported by calculations suggesting that a diet of raw food could not supply sufficient calories for a normal hunter gatherer lifestyle. In particular, many plant foods are too fiber-rich when raw, while most raw meat appears too tough to allow easy chewing. If cooking is indeed obligatory for humans but not for other apes, this means that human biology must have adapted to the ingestion of cooked food (i.e. food that is tender and low in fiber) in ways that no longer allow efficient processing of raw foods. Cooking has been practiced for ample time to allow the evolution of such adaptations. Digestive adaptations have not been investigated in detail but may include small teeth, small hind-guts, large small intestines, a fast gut passage rate, and possibly reduced ability to detoxify. The adoption of cooking can also be expected to have had far-reaching effects on such aspects of human biology as life-history, social behavior, and evolutionary psychology. Since dietary adaptations are central to understanding species evolution, cooking appears to have been a key feature of the environment of human evolutionary adaptedness. Further investigation is therefore needed of the ways in which human digestive physiology is constrained by the need for food of relatively high caloric density compared to other great apes. PMID- 14527629 TI - Micronutrient intakes of wild primates: are humans different? AB - Low micronutrient intake is implicated in a diversity of human health problems, ranging from problems associated with food insufficiency to those associated with food over-consumption. Humans are members of the order primates, suborder anthropoidea, and are most closely related to the great apes. Humans and apes are remarkably similar biologically. In the wild, apes and monkeys consume diets composed largely of plant foods, primarily the fruits and leaves of tropical forest trees and vines. Considerable evidence indicates that the ancestral line giving rise to humans (Homo spp.) was likewise strongly herbivorous (plant eating). The wild plant parts consumed by apes and monkeys show moderate to high levels of many minerals and vitamins. The estimated daily intake of specific minerals, vitamin C and some other vitamins by wild primates is often quite high in comparison to intake levels of these same micronutrients recommended for humans. Are the high micronutrient intakes of wild primates simply a non functional, unavoidable by-product of their strongly plant-based diets or might they actually be serving important as yet undetermined immunological or other beneficial functions? A better understanding of the basis for this apparent difference between humans and wild primates could help to clarify the range and proportions of micronutrients best suited for optimal human development, health and longevity. PMID- 14527630 TI - Dietary diversity, child nutrition and health in contemporary African communities. AB - Many infants in sub-Saharan Africa (SSA) begin to receive cereal-based supplemental feeds well before the age (6 months) recommended for the introduction of 'safe and nutritionally adequate' complementary foods, or in rarer instances, do not receive these until the second year. The diets offered are monotonous and bulky, and rarely cover the shortfall left by breast milk in providing the energy and nutrients required to support rapid growth, build nutrient stores and assure resistance to infection. The pattern of growth and prevalence of malnutrition observed from birth through the first 5 years in SSA are suggestive of the nutrient inadequacies of the diet and the experience of infection. However, it is difficult to link poor growth and specific nutrient deficiencies in epidemiological studies because multiple nutrients are required for growth and deficiencies usually involve several nutrients. Moreover, accurate measurement of nutrient intakes is no small challenge. In this regard, qualitative and easier-to-measure characteristics of diet which are associated with nutrient adequacy could serve as alternative determinant factors in studies looking at causes of malnutrition. Dietary diversity is proposed as a candidate indicator of food security and predictor of nutritional status, but there is need for further research to standardize definitions and methodology before it can be applied widely. PMID- 14527631 TI - Rapid morphological change in living humans: implications for modern human origins. AB - Human body size and body proportions are interpreted as markers of ethnicity, 'race,' adaptation to temperature, nutritional history and socioeconomic status. Some studies emphasize only one of these indicators and other studies consider combinations of indicators. To better understand the biocultural nature of human size and proportions a new study of the growth of Maya-American youngsters was undertaken in 1999 and 2000. One purpose of this research is to assess changes in body proportion between Maya growing up in the US and Maya growing up in Guatemala. Height and sitting height of 6-12-year-old boys and girls (n=360) were measured and the sitting height ratio [sitting height/height]x100, a measure of proportion, was calculated. These data are compared with a sample of Maya of the same ages living in Guatemala and measured in 1998 (n=1297). Maya-American children are currently 10.24 cm taller, on average, and have a significantly lower sitting height ratio, (i.e. relatively longer legs, averaging 7.02 cm longer) than the Guatemala Maya. Maya-American children have body proportions more like those of white children in the US than like Maya children in Guatemala. Improvements in the environment for growth, in terms of nutrition and health, seem to explain both the trends in greater stature and relatively longer legs for the Maya-Americans. These findings are applied to the problem of modern human origins as assessed from fossil skeletons. It has been proposed that heat adapted, relatively long-legged Homo sapiens from Africa replaced the cold adapted, relatively short-legged Homo neandertalensis of the Levant and Europe [J Hum Evol 32 (1997a) 423]. Skeletal samples of Maya adults from rural Guatemala have body proportions similar to adult Neandertals and to skeletal samples from Europe with evidence of nutritional and disease stress. Just as nutrition and health status explains the differences in the body proportions of living Maya children, these factors, along with adaptation to climate, may also explain much of the differences between the Neandertal and African hominid samples. PMID- 14527632 TI - Testing evolutionary hypotheses about human biological adaptation using cross cultural comparison. AB - Physiological data from a range of human populations living in different environments can provide valuable information for testing evolutionary hypotheses about human adaptation. By taking into account the effects of population history, phylogenetic comparative methods can help us determine whether variation results from selection due to particular environmental variables. These selective forces could even be due to cultural traits-which means that gene-culture co-evolution may be occurring. In this paper, we outline two examples of the use of these approaches to test adaptive hypotheses that explain global variation in two physiological traits: the first is lactose digestion capacity in adults, and the second is population sex-ratio at birth. We show that lower than average sex ratio at birth is associated with high fertility, and argue that global variation in sex ratio at birth has evolved as a response to the high physiological costs of producing boys in high fertility populations. PMID- 14527633 TI - Hyperinsulinemic diseases of civilization: more than just Syndrome X. AB - Compensatory hyperinsulinemia stemming from peripheral insulin resistance is a well-recognized metabolic disturbance that is at the root cause of diseases and maladies of Syndrome X (hypertension, type 2 diabetes, dyslipidemia, coronary artery disease, obesity, abnormal glucose tolerance). Abnormalities of fibrinolysis and hyperuricemia also appear to be members of the cluster of illnesses comprising Syndrome X. Insulin is a well-established growth-promoting hormone, and recent evidence indicates that hyperinsulinemia causes a shift in a number of endocrine pathways that may favor unregulated tissue growth leading to additional illnesses. Specifically, hyperinsulinemia elevates serum concentrations of free insulin-like growth factor-1 (IGF-1) and androgens, while simultaneously reducing insulin-like growth factor-binding protein 3 (IGFBP-3) and sex hormone-binding globulin (SHBG). Since IGFBP-3 is a ligand for the nuclear retinoid X receptor alpha, insulin-mediated reductions in IGFBP-3 may also influence transcription of anti-proliferative genes normally activated by the body's endogenous retinoids. These endocrine shifts alter cellular proliferation and growth in a variety of tissues, the clinical course of which may promote acne, early menarche, certain epithelial cell carcinomas, increased stature, myopia, cutaneous papillomas (skin tags), acanthosis nigricans, polycystic ovary syndrome (PCOS) and male vertex balding. Consequently, these illnesses and conditions may, in part, have hyperinsulinemia at their root cause and therefore should be classified among the diseases of Syndrome X. PMID- 14527634 TI - Evolution of dietary antioxidants. AB - Oxygen is vital for most organisms but, paradoxically, damages key biological sites. Oxygenic threat is met by antioxidants that evolved in parallel with our oxygenic atmosphere. Plants employ antioxidants to defend their structures against reactive oxygen species (ROS; oxidants) produced during photosynthesis. The human body is exposed to these same oxidants, and we have also evolved an effective antioxidant system. However, this is not infallible. ROS breach defences, oxidative damage ensues, accumulates with age, and causes a variety of pathological changes. Plant-based, antioxidant-rich foods traditionally formed the major part of the human diet, and plant-based dietary antioxidants are hypothesized to have an important role in maintaining human health. This hypothesis is logical in evolutionary terms, especially when we consider the relatively hypoxic environment in which humans may have evolved. In this paper, the human diet is discussed briefly in terms of its evolutionary development, different strategies of antioxidant defence are outlined, and evolution of dietary antioxidants is discussed from the perspectives of plant need and our current dietary requirements. Finally, possibilities in regard to dietary antioxidants, evolution, and human health are presented, and an evolutionary cost benefit analysis is presented in relation to why we lost the ability to make ascorbic acid (vitamin C) although we retained an absolute requirement for it. PMID- 14527635 TI - Fish consumption, blood docosahexaenoic acid and chronic diseases in Chinese rural populations. AB - The Chinese traditional diet is low in fat. However, there is regional variability in the amount, type of fat consumed and the pattern of chronic diseases. An epidemiological survey of 65 rural counties in China (6500 subjects) was conducted in the 1980s. We have re-examined the red blood cell fatty acid and antioxidant composition, with fish consumption. Fish consumption correlated significantly with the levels of docosahexaenoic acid (DHA) in red blood cells (RBC) (r=0.640, P<0.001), selenium (r=0.467, P<0.001) and glutathione peroxidase (r=0.333, P<0.01) in plasma. The proportion of DHA in RBC was inversely associated with total plasma triglyceride concentrations. A strong inverse correlation between DHA in RBC and cardiovascular disease (CVD) was found. The strongest correlation was the combination of DHA and oleic acid. RBC docosahexaenoic acid itself also correlated negatively and significantly with most chronic diseases and appeared to be more protective than either eicosapentaenoic or the omega3 docosapenataenoic acids. These results demonstrate the protective nature of fish consumption and DHA, found in high fat Western diets, operates at a low level of fat. This finding suggests the protective effect of fish consumption as validated by red cell DHA is universal. The protective effect is, therefore, most likely to be due to the fundamental properties of docosahexaenoic acid in cell function. PMID- 14527636 TI - The Garden of Eden--plant based diets, the genetic drive to conserve cholesterol and its implications for heart disease in the 21st century. AB - It is likely that plant food consumption throughout much of human evolution shaped the dietary requirements of contemporary humans. Diets would have been high in dietary fiber, vegetable protein, plant sterols and associated phytochemicals, and low in saturated and trans-fatty acids and other substrates for cholesterol biosynthesis. To meet the body's needs for cholesterol, we believe genetic differences and polymorphisms were conserved by evolution, which tended to raise serum cholesterol levels. As a result modern man, with a radically different diet and lifestyle, especially in middle age, is now recommended to take medications to lower cholesterol and reduce the risk of cardiovascular disease. Experimental introduction of high intakes of viscous fibers, vegetable proteins and plant sterols in the form of a possible Myocene diet of leafy vegetables, fruit and nuts, lowered serum LDL-cholesterol in healthy volunteers by over 30%, equivalent to first generation statins, the standard cholesterol-lowering medications. Furthermore, supplementation of a modern therapeutic diet in hyperlipidemic subjects with the same components taken as oat, barley and psyllium for viscous fibers, soy and almonds for vegetable proteins and plant sterol-enriched margarine produced similar reductions in LDL cholesterol as the Myocene-like diet and reduced the majority of subjects' blood lipids concentrations into the normal range. We conclude that reintroduction of plant food components, which would have been present in large quantities in the plant based diets eaten throughout most of human evolution into modern diets can correct the lipid abnormalities associated with contemporary eating patterns and reduce the need for pharmacological interventions. PMID- 14527637 TI - An evolutionary perspective on human physical activity: implications for health. AB - At present, human genes and human lives are incongruent, especially in affluent Western nations. When our current genome was originally selected, daily physical exertion was obligatory; our biochemistry and physiology are designed to function optimally in such circumstances. However, today's mechanized, technologically oriented conditions allow and even promote an unprecedentedly sedentary lifestyle. Many important health problems are affected by this imbalance, including atherosclerosis, obesity, age-related fractures and diabetes, among others. Most physicians recognize that regular exercise is a critical component of effective health promotion regimens, but there is substantial disagreement about details, most importantly volume: how much daily caloric expenditure, as physical activity, is desirable. Because epidemiology-based recommendations vary, often confusing and alienating the health-conscious public, an independent estimate, arising from a separate scientific discipline, is desirable, at least for purposes of triangulation. The retrojected level of ancestral physical activity might meet this need. The best available such reconstruction suggests that the World Health Organization's recommendation, a physical activity level of 1.75 ( approximately 2.1 MJ (490 kcal)/d), most closely approximates the Paleolithic standard, that for which our genetic makeup was originally selected. PMID- 14527638 TI - Kenyan dominance in distance running. AB - Critical physiological factors for performance in running are maximal oxygen consumption (VO(2max)), fractional VO(2max) utilization and running economy. While Kenyan and Caucasian elite runners are able to reach very high, but similar maximal oxygen uptake levels, the VO(2max) of black South African elite runners seems to be slightly lower. Moreover, the studies of black and white South African runners indicate that the former are able to sustain the highest fraction of VO(2max) during long distance running. Results on adolescent Kenyan and Caucasian boys show that these boys are running at a similar percentage of VO(2max) during competition. Kenyan elite runners, however, appear to be able to run at a high % of VO(2max) which must then have been achieved by training. A lower energy cost of running has been demonstrated in Kenyan elite runners and in untrained adolescent Kenyan boys compared to their Caucasian counterparts. In agreement with this are the results from studies on black South African elite runners who have shown similar low energy costs during running as the Kenyan elite runners. The good running economy cannot be explained by differences in muscle fibre type as they are the same in Kenyan and Caucasian runners. The same is true when comparing untrained adolescent Kenyan boys with their Caucasian counterparts. A difference exists in BMI and body shape, and the Kenyans long, slender legs could be advantageous when running as the energy cost when running is a function of leg mass. Studies comparing the response to training of Kenyans and Caucasians have shown similar trainability with respect to VO(2max), running economy and oxidative enzymes. Taken all these data together it appears that running at a high fractional VO(2max) and having a good running economy may be the primary factors favouring the good performance of endurance athletes rather than them having a higher VO(2max) than other elite runners. In addition to having the proper genes to shape their bodies and thereby contributing to a good running economy, the Kenyan elite runners have trained effectively and used their potential to be in the upper range both in regard to VO(2max) and to a high utilization of this capacity during endurance running. PMID- 14527639 TI - What makes an endurance athlete world-class? Not simply a physiological conundrum. AB - Inter-individual variation in endurance performance capacity is a characteristic, not only of the general population, but also in trained athletes. The ability of sport scientists to predict which athletes amongst an elite group will become world-class is limited. We do not fully understand the interactions between biological factors, training, recovery and competitive performance. Assessment methods and interpretation of results do not take into account the facts that most research is not done on elite athletes and performances of world-class endurance athletes cannot be attributed to aerobic capacity alone. Many lines of evidence suggest that there is a limit to adaptation in aerobic capacity. Recent advances in molecular biology and genetics should be harnessed by exercise biologists in conjunction with previously used physiological, histological and biochemical techniques to study elite athletes and their responses to different training and recovery regimens. Technological advances should be harnessed to study world-class athletes to determine optimal training and competition strategies. In summary, it is likely that multiple factors are essential contributors to world-class endurance performance and that it is only by using a multidisciplinary approach that we will come closer to solving the conundrum: 'What makes an endurance athlete world class?' PMID- 14527640 TI - The search for genotypes that underlie human performance phenotypes. AB - For a species spread throughout the world, humans are remarkably invariant; yet there has always been more interest in the slight differences between individuals than in the great commonality. This is especially true in athletic endeavours, where nearly immeasurable differences in performance can separate the winner from the rest of the competitors. There is little doubt that performance is influenced by environment, as the effects of diet and training on athletic ability have long been known, if not completely understood; however, the contribution of an individual's genetic make-up is less clear. The dominance of particular nationalities, ethnic groups, or families in various sporting events is often perceived as evidence that heritage (biological or cultural), plays a role in the development of athletic skills. Further complicating the issue are the interactions between genetic background and environment, as both of these fundamental arbiters of development rarely act independently. Despite the complexity of the problem, numerous researchers have attempted to elucidate the effects of genetic background on physical performance and, more recently, to identify the specific genetic variants that contribute to performance. This article reviews some of these studies with a focus on the methodologies employed. PMID- 14527641 TI - Diversity in and adaptation to breath-hold diving in humans. AB - Several features of potential adaptation to breath-hold diving in diving populations and extreme divers are reviewed. Thermal adaptation consists of an improvement in cold tolerance, as witnessed by a decrease in critical water temperature, and implies an elevation of the shivering threshold associated with a greater body insulation. This is indicative of either a strong peripheral vasoconstriction or a more effective countercurrent heat exchange. Respiratory adaptation consists of a blunted ventilatory response to carbon dioxide and an enlargement of lung volumes. Finally, the occurrence of a diving response has been demonstrated. An extreme peripheral vasoconstriction is associated with a dramatic increase in arterial blood pressure. The consequent stimulation of arterial baroreceptors causes an extreme drop of heart rate. Bradycardia is not compensated by a higher stroke volume, with consequent decrease in cardiac output. This decrease, however, is not such as to undermine perfusion to vital organs. Redistribution of blood flow occurs, and some organs such as skeletal muscle may become unperfused, as indicated by the high blood lactate concentrations at low metabolic rate. It is not possible to state, however, whether these changes reflect genetic adaptations or an adaptive response to a prolonged environmental stress. PMID- 14527642 TI - Patterns of control of maximum metabolic rate in humans. AB - In this analysis, four performance phenotypes were used to compare mechanisms of control of aerobic maximum metabolic rate (MMR): (i) untrained sedentary (US) subjects, as a reference group against which to compare (ii) power trained (PT), (iii) endurance trained (ET) and (iv) high altitude adapted native (HA) subject groups. Sprinters represented the PT group; long distance runners illustrated the ET group; and Quechuas represented the HA group. Numerous recent studies have identified contributors to control on both the adenosine triphosphate (ATP) supply side and the ATP demand side of ATP turnover. Control coefficients or c(i) values were defined as fractional change in MMR/fractional change in the capacity of any given step in ATP turnover. From the best available evidence it appears that at MMR all five of the major steps in energy delivery (namely, ventilation, pulmonary diffusion, cardiac output, tissue capillary - mitochondrial O(2) transfer, and aerobic cell metabolism per se) approach an upper functional ceiling, with control strength being distributed amongst the various O(2) flux steps. On the energy demand side, the situation is somewhat simplified since at MMR approximately 90% of O(2)-based ATP synthesis is used for actomyosin (AM) and Ca(2+) ATPases; at MMR these two ATP demand rates also appear to be near an upper functional ceiling. In consequence, at MMR the control contributions or c(i) values are rather evenly divided amongst all seven major steps in ATP supply and ATP demand pathways right to the point of fatigue. Relative to US (the reference group), in PT subjects at MMR control strength shifts towards O(2) delivery steps (ventilation, pulmonary diffusion and cardiac output). In contrast in ET and HA subjects at MMR control shifts towards the energy demand steps (AM and Ca(2+) ATPases), and more control strength is focussed on tissue level ATP supply and ATP demand. One obvious advantage of the ET and HA control pattern is improved metabolite homeostasis. Another possibility is that, with some reserve capacity in the O(2) delivery steps and control focussed on ATP turnover at the tissue level, nature has designed the ideal 'endurance machine'. PMID- 14527643 TI - Hypothalamic-pituitary-adrenal axis responses to stress in subjects with 3,4 methylenedioxy-methamphetamine ('ecstasy') use history: correlation with dopamine receptor sensitivity. AB - Fifteen 3,4-methylenedioxymethamphetamine (MDMA, 'ecstasy') users who did not have other drug dependencies or prolonged alcohol abuse and 15 control subjects were studied. All the subjects were exposed to the same psychosocial stressor (Stroop Color-Word Interference Task, public speaking and mental arithmetic in front of an audience) 3 weeks after MDMA discontinuation. Plasma concentrations of adrenocorticotropic hormone (ACTH) and cortisol were measured immediately before the tests began and at their end, 30 min later. Growth hormone (GH) responses to the dopaminergic agonist bromocriptine and psychometric measures (Tridimensional Personality Questionnaire, Minnesota Multiphasic Personality Inventory, Buss-Durkee Hostility Inventory) were also obtained 4 weeks after MDMA discontinuation for the same subjects. ACTH and cortisol basal levels were significantly higher in ecstasy users than in control subjects. In contrast, ACTH and cortisol responses to stress were significantly blunted in MDMA users. The sensitivity of dopamine D2 receptors, reflected by GH responses to bromocriptine challenge, was reduced in MDMA users compared with controls. The responsiveness of the hypothalamic-pituitary-adrenal (HPA) axis (ACTH and cortisol delta peaks) correlated directly with GH areas under curves in response to bromocriptine, and inversely with psychometric measures of aggressiveness and novelty seeking. No correlation was found between hormonal measures and the extent of MDMA exposure. Reduced D2 receptor sensitivity, HPA basal hyperactivation and reduced responsiveness to stress may represent a complex neuroendocrine dysfunction associated with MDMA use. The present findings do not exclude the possibility that dopamine dysfunction partly predated MDMA exposure. PMID- 14527644 TI - Vulnerability to 35% CO2 of panic disorder patients with a history of respiratory disorders. AB - Patients with panic disorder often report a history of respiratory pathology. Furthermore, panic disorder patients are vulnerable to CO2 challenges. The increased CO2 vulnerability displayed by panic disorder patients may be related to lifetime respiratory pathology. We examined whether panic disorder patients with a history of respiratory disorders are more vulnerable to a 35% CO2 challenge than those without such a history. Ninety-six patients with panic disorder were interviewed about their lifetime respiratory status (asthma, bronchitis and various other respiratory conditions) and underwent the challenge. Immediately before and after the CO2 inhalation, the patients filled out the Visual Analogue Scale for Anxiety (VAS-A) and the Panic Symptom List (PSL). We found no differences between the two panic disorder groups on anxiety (VAS-A), panic symptoms (PSL) or panic attacks after the CO2 challenge. Our results suggest that having a PD is an important factor in CO2 vulnerability independent of a history of respiratory disorders. PMID- 14527645 TI - Automatic processing of verbal emotion stimuli in schizophrenia. AB - In the present study, automatic processing of verbal emotion stimuli was investigated as a function of affective symptoms, emotional state and trait characteristics of chronic schizophrenia patients. A sequential pronunciation priming task was administered to 30 schizophrenia patients with a flat affect expression, 30 schizophrenia patients suffering from anhedonia, 28 schizophrenia patients not suffering from anhedonia or flat affect, and 30 healthy subjects to assess affective and semantic priming effects. The Scale for the Assessment of Negative Symptoms (Andreasen, British Journal of Psychiatry, 1989, 155, 53-58) was applied to evaluate the flat affect and anhedonia, and to categorize patients into groups. Schizophrenia patients did not differ from healthy subjects in affective and semantic priming scores. However, affective priming based on positive primes was inversely correlated with negative state and trait affectivity, and positively correlated with trait joy in the patient sample. These results support the view that a decrement in automatic processing facilitation of positive valence might play a role in the development of negative emotions. The symptoms of flat affect and anhedonia do not appear to be associated with impairments in the automatic processing of verbal emotional material in schizophrenia. PMID- 14527646 TI - Executive function assessed by memory updating and random generation in schizotypal individuals. AB - Individuals scoring high (N=32) and low (N=27) on the unusual experiences (UnEx) scale of the Oxford-Liverpool Inventory of Feelings and Experiences (O-LIFE) were selected from a large sample (N=265) of normal volunteer undergraduates. The high and low-UnEx groups were compared on two tasks, random generation and memory updating, which target executive functions that inhibit prepotent responses and update current information. The groups differed only on the R measure of random generation that assesses inequality in the relative frequencies of response alternatives, a result attributed to superstitious behaviour rather than to executive deficit. The results suggest that the executive impairments previously observed in high schizotypal individuals using the Wisconsin Card Sorting Test and other measures are selective rather than global. PMID- 14527647 TI - A brief smell identification test discriminates between deficit and non-deficit schizophrenia. AB - Evidence is accumulating that smell identification deficits (SID) and social dysfunction in schizophrenia may share a common pathophysiology. While most schizophrenia studies utilize the lengthy 40-item University of Pennsylvania Smell Identification Test (UPSIT) to assess smell identification ability, a brief 12-item smell identification test (B-SIT) has recently been constructed as a culturally neutral substitute for the UPSIT. By selecting the 12 items of the UPSIT from which the B-SIT was originally derived, we constructed a proxy for the B-SIT and compared the performance of 83 patients with schizophrenia to 69 normal subjects. We examined select properties of the B-SIT proxy in relation to the UPSIT to determine its efficacy for use in psychiatric populations. We considered the sensitivity of the B-SIT proxy and evaluated a cutoff score for identifying deficit syndrome schizophrenia (DS). The UPSIT and B-SIT proxy were significantly related in the patients (n=83, r=0.85, P=0.01) and in comparison subjects (n=69, r=0.83, P=0.01), and both measures similarly distinguished DS from non-deficit syndrome (non-DS) patients. The results of this study support the utility of the B-SIT for schizophrenia research and highlight the robustness of the relationship between SID and social dysfunction in schizophrenia. PMID- 14527648 TI - Portable mood mapping: the validity and reliability of analog scale displays for mood assessment via hand-held computer. AB - The long-term natural time course of mood change remains poorly understood, and improved methods that assay multiple mood symptoms quickly and reliably are crucial to further progress. This study describes the reliability and validity of the new visual analog scale (VAS) display method for a recently developed 19-item VAS-based mood questionnaire, the VMQ, administered via hand-held computer (HHC). The effect of the smaller HHC screen size on accuracy and precision of VAS completion was investigated in 28 subjects using 4- and 10-cm paper-based VASs to indicate six specified dates within the year. The influence of digital vs. paper medium was then tested in 39 subjects who completed the same task, using 10-cm paper and 4-cm HHC-based VASs. Test-retest reliability was evaluated in 29 subjects who completed the questionnaire on a HHC twice, 10 min apart. Since the HHC presents VMQ scales with text anchor orientation set randomly, we also considered whether subjects might inadvertently transpose responses on the HHC. We found that reducing VAS size produced no significant loss of response precision or accuracy in subject response. Moreover, there was no significant loss of accuracy or precision between 10-cm paper and 4-cm HHC-based versions of the VAS. HHC-based items also demonstrated excellent test-retest reliability, with excellent values of Cronbach's alpha. The transposition error rate was negligible (0.27%). Our study provides initial evidence that the HHC-based VAS display used in the VMQ is a reliable and valid tool for comprehensive collection of analog mood scale data. PMID- 14527649 TI - Correlation of subjective and objective sleep measurements at different stages of the treatment of depression. AB - Studies of the correlation of subjective and objective sleep measures in depressed patients have produced mixed results so far. Further, they were carried out in sleep laboratories and tended to obtain one-off assessments, thus not taking into account the effect of treatment. We investigated forty (40) patients over the course of 8-week treatment of depression with either paroxetine or nefazodone. We used home polysomnography at baseline, nights 3 and 10, and week 8 of treatment, with extensive assessments of subjective sleep, the morning after each sleep recording. The patients were able to judge accurately their total sleep time and sleep onset latency, both before and during treatment. However, they were inaccurate in estimating the number of times they woke up during the night. Sleep satisfaction correlated negatively with Stage 1 sleep at baseline. Sleep quality was represented by a combination of subjective parameters measuring the ease of initiation and maintenance of sleep, and it appeared to derive from slow wave sleep and sleep continuity as seen in polysomnography. The partial discrepancy between subjective and objective measures suggests that a cognitive element is combined with the biological element to produce the sleep problems reported by depressed patients. PMID- 14527650 TI - First-night effect in the chronic fatigue syndrome. AB - Since the magnitude of the first-night effect has been shown to be a function of medical conditions and of settings in which polysomnographies are performed, it is essential to evaluate the habituation phenomenon in each case in order to determine the optimal recording methodology. A first-night effect was evidenced in certain cases of chronic fatigue syndrome, but not in others. To clarify this issue, a large group of patients with chronic fatigue syndrome who had no primary sleep disorders were selected and recorded for two consecutive nights in a hospital sleep unit. Several parameters, frequently associated with the first night effect, were found to be influenced by the recording methodology: Total Sleep Time, Sleep Efficiency, Sleep Efficiency minus Sleep Onset, Sleep Onset Latency, Wake Time, Slow Wave Sleep, Rapid Eye Movement Sleep, Rapid Eye Movement Sleep Latency and Number of Sleep Cycles. Bland and Altman plots determined that the difference scores between the nights included a systematic bias linked to the order of recordings (first-night effect). Factorial analysis grouped the difference scores into three factors. No significant difference was observed between patients with generalized anxiety comorbidity and those with no psychiatric comorbidity, or between those with and without psychiatric comorbidity. Chronic fatigue syndrome must thus be added on the list of conditions where a clinically significant habituation effect takes place. PMID- 14527651 TI - Detection of anti-Borna Disease Virus (BDV) antibodies from patients with schizophrenia and mood disorders in Japan. AB - The relationship between infection with the Borna Disease Virus (BDV) and the clinical symptoms of schizophrenia and mood disorders (DMS-IV) was investigated. Western blotting techniques were used to examine anti-p10-BDV antibodies in serum from 32 patients with schizophrenia and 33 patients with mood disorders in Japan. The results showed that 1 out of 25 controls (4.0%), 7 out of 32 patients with schizophrenia (21.9%) and 9 out of 33 patients with mood disorders (27.3%) were positive for anti-BDV-p10 antibodies. Compared with levels of anti-BDV-p10 antibodies in controls, the production of anti-BDV-p10 antibodies failed to show a statistically significant relationship with schizophrenia but did show a significant relationship with mood disorder. The subgroup of schizophrenia patients with positive syndromes had a non-significantly higher frequency of anti BDV-p10 antibodies than the subgroup of patients with negative syndromes. Similarly, the production of anti-BDV-p10 antibodies was non-significantly higher among patients with the unipolar subtype of mood disorder than in those with the bipolar subtype. PMID- 14527652 TI - Smoking habits in Japanese patients with schizophrenia. AB - Studies from North America and Western Europe have observed a marked increase of smoking in schizophrenia. This preliminary study investigated smoking habits in Japanese patients with schizophrenia (n=137). The prevalence of smokers (34%) was not higher than in the general Japanese population (37%). Variables associated with smoking were also different from those reported in the Western literature. Different cultural backgrounds may underlie the differing results in Western and Japanese populations. PMID- 14527653 TI - DNA repair by bacterial AlkB proteins. AB - The Escherichia coli AlkB protein is an iron- and 2-oxoglutarate-dependent oxygenase, repairing 1-methyladenine and 3-methylcytosine lesions in DNA. AlkB homologues are present in a number of bacterial species, and some bacteria have two different AlkB proteins. AlkB also repairs lesions in RNA, and the biological significance of RNA repair is discussed. PMID- 14527654 TI - Conidiation induction in Penicillium. AB - Asexual spores or conidia are dispersive propagules produced as an alternative to vegetative growth by a diverse group of filamentous fungi. The cellular development programmes which govern conidiation have been intensely studied in the last few decades, although important gaps stand in the way of our understanding of this phenomenon, namely in the areas of the environmental sensing mechanisms and signal transduction pathways. The aim of this review is to summarize the current advances in conidiation induction in the genus Penicillium, and to put them into context with the state of our knowledge stemming from work in widely studied fungal model systems. PMID- 14527655 TI - A role for bacteriophage T4 rI gene function in the control of phage development during pseudolysogeny and in slowly growing host cells. AB - Although most studies on bacteriophages have been performed under laboratory conditions that are optimal for host cell growth, in nature, bacteria and bacteriophages coexist in different habitats. Here, by using different growth rates in carbon-limited chemostats, we investigated the development of phage T4 in its host Escherichia coli. Our results strongly suggest that T4 can form pseudolysogens not only when bacterial growth is completely inhibited, but also in growing host cells. The rI gene, previously known to be indispensable for lysis inhibition, seems to play an important role in optimization of phage development in slowly growing cells as well as during establishment and maintenance of pseudolysogeny. PMID- 14527656 TI - DNA sequence and analysis of the pYVa127/90 virulence plasmid of Yersinia enterocolitica strain A127/90. AB - The complete nucleotide sequence and organization of the Yersinia enterocolitica strain A127/90 serotype O:8 biotype 1B virulence plasmid (pYVa127/90) were determined. In contrast to other biotype 1B strains, usually isolated in the USA, strain A127/90 was isolated in Japan. The organization of the genes carried by the 66591-bp pYVa127/90 plasmid is globally similar to that of 67720-bp pYVe8081, the virulence plasmid from 8081, an American isolate of serotype O:8 biotype 1B. Differences are noticeable in the amino acid sequences of the SycH chaperone, the YopM cytotoxin and the plague-protective antigen LcrV. Interestingly, pYVa127/90 contains the entire coding sequence of the YlpA lipoprotein, which is missing from pYVe8081. In addition, we identified five potential new genes. Several vestigial insertion sequence elements appeared to be different between the two plasmids. These observations suggest that both pYV plasmids are from a common ancestor but may have evolved independently following the conquest of two distant ecological niches. PMID- 14527657 TI - DNA array analysis of gene expression in response to UV irradiation in Escherichia coli. AB - The capacity of DNA macroarrays that contain all 4290 predicted open reading frames of the E. coli K12 genome was evaluated by measuring changes in gene expression in response to irradiation by ultraviolet light (UV). UV and other DNA damaging agents are known to trigger the induction of the SOS response. This is a coordinated increase in the level of expression of a set of approximately 30 unlinked genes, the SOS genes, negatively regulated by the LexA repressor. The analysis was performed on a set of isogenic strains with mutations that affect expression of genes of the SOS system: (i) the lexA+ strain, in which the SOS system can be induced after DNA damage, (ii) lexAind- mutants in which the SOS system cannot be induced, and (iii) lexAdef mutants in which the SOS system is induced constitutively. We found that a large set of genes appeared to be either upregulated or downregulated following UV irradiation. Among the genes which appeared to be upregulated in a LexA-dependent manner, we correctly identified 9 out of 27 SOS genes printed on the arrays and one gene containing a LexA binding site. One gene, dnaN, encoding the beta subunit of DNA polymerase III holoenzyme, was identified as an upregulated gene in a LexA-independent manner. Our results were compared to those of similar studies previously published. Although the SOS response as a whole could not be illustrated by using DNA arrays, the data suggest that regulation of some SOS genes might be more complex than previously thought. PMID- 14527658 TI - Changes in Escherichia coli transcriptome during acclimatization at low temperature. AB - Upon cold shock Escherichia coli transiently stops growing and adapts to the new temperature (acclimatization phase). The major physiological effects of cold temperature are a decrease in membrane fluidity and the stabilization of secondary structures of RNA and DNA, which may affect the efficiencies of translation, transcription, and replication. Specific proteins are transiently induced in the acclimatization phase. mRNA stabilization and increased translatability play a major role in this phenomenon. Polynucleotide phosphorylase (PNPase) is one of the cold-induced proteins and is essential for E. coli growth at low temperatures. We investigated the global changes in mRNA abundance during cold adaptation both in wild type E. coli MG1655 and in a PNPase deficient mutant. We observed a twofold or greater variation in the relative mRNA abundance of 20 genes upon cold shock, notably the cold-inducible subset of csp genes and genes not previously associated with cold shock response, among these, the extracytoplasmic stress response regulators rpoE and rseA, and eight genes with unknown function. Interestingly, we found that PNPase both negatively and positively modulated the transcript abundance of some of these genes, thus suggesting a complex role of PNPase in controlling cold adaptation. PMID- 14527659 TI - Direct detection of leptospiral material in human postmortem samples. AB - Leptospiral culture, direct immunofluorescence, and the polymerase chain reaction (PCR) were used to detect leptospiral material in postmortem specimens collected from eight patients who died of leptospirosis. Diagnosis of leptospiral infection was based on clinical summary (premortem) and confirmed by serological analysis and/or culture of leptospires. Leptospiral culture was the least sensitive technique, yielding two isolates (3%) from 65 samples. Both isolates were from the aqueous humour and cerebrospinal fluid of the same patient. Direct immunofluorescence was of intermediate sensitivity for detection of leptospires, confirming the presence of leptospires in 11% (2 of 18) of tissue samples from three patients. PCR analysis was the most sensitive technique for detection of leptospiral material in tissue samples, being positive in 20% (11 of 56) of samples from eight patients. Both samples (cerebellum and liver) positive by immunofluorescence were also positive by PCR. The sensitivity of the PCR assay was 1-10 leptospires ml(-1) sample, and the assay was specific for Leptospira pathogenic species. Multi-system involvement was indicated based on successful amplification of leptospiral DNA from more than one tissue sample, which corroborated with the clinical and pathologic findings. The results suggest that in acute and/or fatal leptospirosis, the pathogenesis of the pathologic features are related to the presence of the organisms in the tissues. In conclusion, PCR combined with serology appears to be a useful tool for diagnosis of leptospirosis and may be invaluable in epidemiological studies. PMID- 14527660 TI - Identification of Klebsiella oxytoca using a specific PCR assay targeting the polygalacturonase pehX gene. AB - Bacteria of the genus Klebsiella are important opportunistic pathogens responsible for nosocomial infections that are increasingly resistant to antimicrobial agents. Distinctive identification of the species K. oxytoca, K. pneumoniae, K. planticola, K. ornithinolytica and K. terrigena is difficult based on phenotypic tests and misidentifications are frequent in routine clinical microbiology. We developed a specific method to discriminate K. oxytoca from the other species of the genus Klebsiella, based on the PCR amplification of the polygalacturonase (pehX) gene. A PCR amplicon of 344 bp was obtained in all 35 K. oxytoca strains tested, but in none of the 29 K. pneumoniae, 12 K. planticola/K. ornithinolytica and 7 K. terrigena strains tested. The test was also negative for polygalacturonate-degrading species of the genus Erwinia. Analysis of 24 strains designated as K. pneumoniae from international collections (NCTC, PZH) revealed previous misidentification of six K. oxytoca strains. Key biochemical tests fully confirmed the pehX PCR results. The new K. oxytoca identification assay should be useful for both clinical and ecological monitoring of K. oxytoca strains, as well as for controlling the previous identification of collection strains. PMID- 14527661 TI - Diversity of polyketide synthase gene sequences in Aspergillus species. AB - Fungal polyketide synthases are responsible for the biosynthesis of several mycotoxins and other secondary metabolites. The aim of our work was to investigate the diversity of polyketide synthases in Aspergillus species using two approaches: PCR amplification using oligonucleotide primers, and bioinformatics. Ketosynthase domain probes amplified DNA fragments of about 700 bp in each examined isolate. Sequences of these domains were aligned and analyzed by phylogenetic methods. The ketosynthase domain sequences were highly diverse indicating that they most probably represent polyketide synthases responsible for different functions. A. albertensis and A. niger ketosynthase domain sequences clustered together with sequences of genes required for pigment biosynthesis (wA) in A. nidulans and P. patulum, while the ketosynthase domain sequence of A. muricatus was most closely related to an A. parasiticus wA type domain sequence, and those of the A. ochraceus isolates formed a distinct clade on the tree. These sequences were highly homologous to an A. terreus naphthopyrone synthase gene. An Aspergillus fumigatus genomic database was also searched for ketosynthase domain sequences, which have been included in the phylogenetic analysis. Altogether 14 putative ketosynthase domain sequences were identified. Clustering of the ketosynthase domain sequences correlated well with the type of metabolites produced by the corresponding polyketide synthases. At least 8 clusters with putative ketosynthase domain sequences of unknown function have been identified. Further studies are in progress to clarify the role of some of the identified polyketide synthase genes. PMID- 14527663 TI - The gelsolin family of actin regulatory proteins: modular structures, versatile functions. AB - This issue of FEBS Letters includes two manuscripts describing structural studies of gelsolin, the best-characterized member of a superfamily of actin binding proteins that sever, cap, and in some cases nucleate and bundle actin filaments. The manuscripts by Narayan et al. and Irobi et al. provide snapshots of gelsolin domains activated by calcium and in complex with the actin monomer, revealing new insights into the remarkable actin regulatory activities of this versatile protein. These studies build upon nearly a quarter of a century of research on gelsolin's effects on actin dynamics and its role in normal and diseased cells. In the following minireview, we summarize the structural studies that have provided insights into gelsolin's severing and capping activities and look to the future of work on this remarkable molecule. PMID- 14527664 TI - Activation in isolation: exposure of the actin-binding site in the C-terminal half of gelsolin does not require actin. AB - Gelsolin requires activation to carry out its severing and capping activities on F-actin. Here, we present the structure of the isolated C-terminal half of gelsolin (G4-G6) at 2.0 A resolution in the presence of Ca(2+) ions. This structure completes a triptych of the states of activation of G4-G6 that illuminates its role in the function of gelsolin. Activated G4-G6 displays an open conformation, with the actin-binding site on G4 fully exposed and all three type-2 Ca(2+) sites occupied. Neither actin nor the type-l Ca(2+), which normally is sandwiched between actin and G4, is required to achieve this conformation. PMID- 14527665 TI - From the first to the second domain of gelsolin: a common path on the surface of actin? AB - We present the 2.6 A resolution crystal structure of a complex formed between G actin and gelsolin fragment Met25-Gln160 (G1+). The structure differs from those of other gelsolin domain 1 (G1) complexes in that an additional six amino acid residues from the crucial linker region into gelsolin domain 2 (G2) are visible and are attached securely to the surface of actin. The linker segment extends away from G1 up the face of actin in a direction that infers G2 will bind along the same long-pitch helical strand as the actin bound to G1. This is consistent with a mechanism whereby G2 attaches gelsolin to the side of a filament and then directs G1 toward a position where it would disrupt actin-actin contacts. Alignment of the sequence of the structurally important residues within the G1-G2 linker with those of WH2 (WASp homology domain 2) domain protein family members (e.g. WASp (Wiscott-Aldridge syndrome protein) and thymosin beta4) suggests that the opposing activities of filament assembly and disassembly may exploit a common patch on the surface of actin. PMID- 14527666 TI - Is there an evolutionary relationship between WARP (von Willebrand factor A domain-related protein) and the FACIT and FACIT-like collagens? AB - We suggest that there is an evolutionary relationship between von Willebrand factor A-domain-related protein (WARP), and the fibril-associated collagen with interrupted triple helix (FACIT) and FACIT-like subfamilies of collagens. Data from a comparison of amino acid sequences, domain organisation and chromosomal location are consistent with the hypothesis that WARP and these collagens share a common collagen ancestor. In support of this is the observation that the WARP 3' coding region is GC-rich suggesting that this may represent the remnant of a triple helix protein domain which WARP has 'lost' during evolution. PMID- 14527667 TI - D-Amino acids and D-Tyr-tRNA(Tyr) deacylase: stereospecificity of the translation machine revisited. AB - Until 30 years ago, it had been considered that D-amino acids were excluded from living systems except for D-amino acids in the cell wall of microorganisms. However, D-amino acids, in the form of free amino acids, peptides and proteins, were recently detected in various living organisms from bacteria to mammals. The extensive distribution of bio-functional D-amino acids challenges the current concept of protein synthesis: more attention should be paid to the stereospecificity of the translation machine. Besides aminoacyl-tRNA synthetases, elongation factor Tu and some other mechanisms, D-Tyr-tRNA(Tyr) deacylases provide a novel checkpoint since they specifically recycle misaminoacylated D-Tyr tRNA(Tyr) and some other D-aminoacyl-tRNAs. Their unique structure represents a new class of tRNA-dependent hydrolase. These unexpected findings have far reaching implications for our understanding of protein synthesis and its origin. PMID- 14527668 TI - The diversity of globin-coupled sensors. AB - The recently discovered globin-coupled sensors (GCSs) are heme-containing two domain transducers distinct from the PAS domain superfamily. We have identified an additional 22 GCSs with varying multi-domain C-terminal transmitters through a search of the complete and incomplete microbial genome datasets. The GCS superfamily is composed of two major subfamilies: the aerotactic and gene regulators. We postulate the existence of protoglobin in Archaea as the predecessor to the chimeric GCS. PMID- 14527669 TI - Ghrelin, a widespread hormone: insights into molecular and cellular regulation of its expression and mechanism of action. AB - The recent discovery of ghrelin, the endogenous ligand for the growth hormone secretagogue receptor, is the result of almost 25 years of research by many groups all around the world, and represents a milestone in our understanding of growth hormone secretion and energy homeostasis. This minireview is focused on recent studies on ghrelin, pointing out the cellular and molecular mechanisms involved in the gene expression of ghrelin since recent studies have unequivocally shown that ghrelin biological activity is dependent on a peculiar post-translational processing. Major interest in this peptide derived from the fact that, in addition to other effects, it is involved in the regulation of energy balance by inducing weight gain and reducing fat utilization. These activities are likely mediated by a CNS network of cells that is also modulated by other hormones such as leptin. Ghrelin has emerged as a premeal initiation factor that informs CNS about the status of the energy balance. The development of ghrelin analogs, agonists and antagonists, appears as a suitable approach for possible therapeutic intervention in a variety of disease states linked to alterations in body weight homeostasis. PMID- 14527670 TI - Rapidly maturing red fluorescent protein variants with strongly enhanced brightness in bacteria. AB - A rapidly maturing variant of the red fluorescent protein DsRed was optimized for bacterial expression by random mutagenesis. The brightest variant contains six mutations, two of which (S4T and a silent mutation in codon 2) explain most of the fluorescence enhancement. The novel variants are expressed at 9-60-fold higher levels in Escherichia coli compared to DsRed.T3, but are not superior fluorophores on a per molecule basis. In contrast to previously available DsRed variants, DsRed.T3_S4T is sufficiently bright to monitor Salmonella gene expression in infected animals using flow cytometry. However, no fluorescence enhancement was observed in Leishmania or HeLa cells, indicating that these novel variants are specifically useful for bacteria. PMID- 14527671 TI - Functional involvement of cathepsin W in the cytotoxic activity of NK-92 cells. AB - Human cathepsin W (lymphopain) is a papain-like cysteine protease of unknown function that is specifically expressed in natural killer (NK) cells and to a lesser extent in cytotoxic T cells (CTL). In order to analyze the functional importance of cathepsin W for the cytotoxic process, we investigated NK-92 cells that have an NK cell-like phenotype and express cathepsin W. NK-92 cells possess strong cytotoxic activity against Jurkat and K562 cells. The cytotoxic activity of NK-92 cells against K562 was decreased in the presence of antisense phosphorothioate oligonucleotides against the cathepsin W-cDNA. Western blot analysis showed that the impaired cytotoxic activity of NK-92 cells was accompanied by reduced amounts of cathepsin W in the antisense-treated cells. In addition, co-cultivation experiments between NK-92 and K562 cells revealed a time dependent decrease of cathepsin W by Western blot and immunofluorescence analysis during the cytotoxic attack, whereas CD56 expression of NK-92 cells was not affected. During cytotoxic attack, cathepsin W was neither targeted to K562 cells or other subcellular compartments, as shown by immunofluorescence analysis. The decrease of cathepsin W protein was associated with stable cathepsin W transcript levels. Control experiments using HT-29 cells, which are resistant against NK-92 mediated cytotoxicity, showed no change of cathepsin W expression, implying that the decrease of cathepsin W in the NK-92/K562 assay is linked to the cytotoxic process. Although the exact function of cathepsin W with respect to its enzymatic activity and its site of action still needs to be elucidated, our data demonstrate for the first time that cathepsin W is important for cellular cytotoxicity mediated by NK cells. PMID- 14527672 TI - Placental leucine aminopeptidase/oxytocinase gene regulation by activator protein 2 in BeWo cell model of human trophoblast differentiation. AB - Placental leucine aminopeptidase (P-LAP) is located preferentially in syncytiotrophoblasts in human placenta. Here we investigated P-LAP expression and the regulatory mechanisms in BeWo choriocarcinoma cells with forskolin (FSK) induced differentiation. Morphologically differentiated cells revealed enhanced P LAP staining. FSK significantly increased P-LAP activity and mRNA. Deletion or mutation of activator protein-2 (AP-2) binding site in the footprint-3 (-216 to 172) of P-LAP promoter abrogated the stimulatory effects of FSK on luciferase activity of the construct -216/+49. In AP-2-deficient Hep-G2 cells, FSK failed to stimulate luciferase activity of the construct -216/+49. Among the isoforms, BeWo expressed AP-2alpha and AP-2gamma, while FSK increased only AP-2alpha. These results suggest differentiation-dependent P-LAP expression in trophoblasts, which involves increased AP-2alpha binding. PMID- 14527673 TI - Changes in Hsp90 expression determine the effects of cyclosporine A on the NO pathway in rat myocardium. AB - Cyclosporine A (CsA) is associated with the development of cardiovascular toxicity in transplant patients but can exert myocardial protection against ischemia/reperfusion damages. We examined in a rat model of chronic CsA administration whether subtle variations in the NO pathway could account for these opposite effects. CsA treatment rapidly led to an increase in myocardial Hsp90 expression promoting the recruitment of Akt and calcineurin, thereby promoting eNOS activation through Ser1177 phosphorylation and Thr495 dephosphorylation, respectively. This was associated with an increase in myocardial VEGF expression and led to anti-apoptotic effects in isolated cardiac myocytes. Upon longer CsA exposure, cardiac toxicity developed, as documented by the infiltration of connective tissue and the increase in iNOS expression. These later effects were associated with a dramatic decrease in the abundance and scaffold function of Hsp90, thereby unraveling the key role of Hsp90 in governing CsA effects. PMID- 14527674 TI - Platelet-derived growth factor induces p21/WAF1 promoter in vascular smooth muscle cells via activation of an Sp1 site. AB - Many studies suggested that cyclin-dependent kinase inhibitor (CDKI) p21 acts as a universal inhibitor of cyclin/CDK catalytic activity. This protein has also been shown to be a component of active cyclin/CDK complexes. In addition, it has recently been suggested that p21 serves as an assembly factor in platelet-derived growth factor (PDGF)-stimulated vascular smooth muscle cells (VSMC). However, little is known concerning the molecular mechanisms by which PDGF induces p21 gene expression in VSMC. In this report we demonstrate that PDGF induces the p21 expression at both the mRNA and protein levels. This increase in p21 gene expression was due to activation of the p21 promoter by PDGF. Through both deletion and mutation analysis of the p21 promoter, we defined a 10-bp sequence that is required for the activation of the p21 promoter by PDGF. In addition, gel shift and supershift assays demonstrated that this PDGF-responsive element binds specifically to the transcription factor Sp1. These results demonstrate that Sp1 mediates PDGF-induced p21 gene expression in VSMC. Moreover, immunoblot and immunoprecipitation analysis showed that the level of hyperphosphorylated retinoblastoma protein (Rb) is increased and the protein is physically associated with Sp1 in PDGF-treated cells, indicating that phosphorylated Rb may play a role in regulating Sp1 to activate p21 expression. PMID- 14527675 TI - Distal side tryptophan, tyrosine and methionine in catalase-peroxidases are covalently linked in solution. AB - Distal side tryptophan and tyrosine have been shown to be essential in the catalase but not the peroxidase activity of bifunctional catalase-peroxidases (KatGs). Recently published crystal structures suggest that both residues could be part of a novel adduct including in addition a conserved methionine. A mass spectrometric analysis of the tryptic peptides from recombinant wild-type Synechocystis KatG and the variants Trp122Phe, Tyr249Phe and Met275Ile confirms that this novel adduct really exists in solution and thus may be common to all KatGs. Exchange of either Trp122 or Tyr249 prevents cross-linking, whereas exchange of Met275 still allowed bond formation between Trp122 and Tyr249. It is proposed that the covalent bond between Trp and Tyr may form before that between Tyr and Met. The findings are discussed with respect to the mechanism of cross linking and its role in KatG catalysis. PMID- 14527676 TI - Artemisinin inhibits inducible nitric oxide synthase and nuclear factor NF-kB activation. AB - Artemisinin is a natural product used as an alternative drug in the treatment of severe and multidrug-resistant malaria. In the present work we show that artemisinin shares with other sesquiterpene lactones the ability to inhibit the activation of the nuclear factor NF-kB: by this mechanism, artemisinin, as well as parthenolide, inhibits nitric oxide synthesis in cytokine-stimulated human astrocytoma T67 cells. These results suggest that artemisinin, in addition to its antiparasitic properties, could also exert a therapeutic effect on neurological complications of malaria. PMID- 14527677 TI - Enhanced in vivo delivery of antisense oligonucleotides to restore dystrophin expression in adult mdx mouse muscle. AB - The use of antisense oligonucleotides (AOs) to induce exon skipping leading to generation of an in-frame dystrophin protein product could be of benefit in around 70% of Duchenne muscular dystrophy patients. We describe the use of hyaluronidase enhanced electrotransfer to deliver uncomplexed 2'-O-methyl modified phosphorothioate AO to adult dystrophic mouse muscle, resulting in dystrophin expression in 20-30% of fibres in tibialis anterior muscle after a single injection. Although expression was transient, many of the corrected fibres initially showed levels of dystrophin expression well above the 20% of endogenous previously shown to be necessary for phenotypic correction of the dystrophic phenotype. PMID- 14527678 TI - Structure-based mutational analyses in FGF7 identify new residues involved in specific interaction with FGFR2IIIb. AB - Receptor binding specificity is an essential element in regulating the diverse activities of fibroblast growth factors (FGFs). FGF7 is ideal to study how this specificity is conferred at the structural level, as it interacts exclusively with one isoform of the FGF-receptor (FGFR) family, known as FGFR2IIIb. Previous mutational analysis suggested the importance of the beta4/beta5 loop of FGF7 in specific receptor recognition. Here a theoretical model of FGFR2IIIb/FGF7 complex showed that this loop interacts with the FGFR2IIIb unique exon. In addition, the model revealed new residues that either directly interact with the FGFR2IIIb unique exon (Asp63, Leu142) or facilitate this interaction (Arg65). Mutations in these residues reduced both receptor binding affinity and biological activity of FGF7. Altogether, these results provide the basis for understanding how receptor binding specificity of FGF7 is conferred at the structural level. PMID- 14527679 TI - Adhesion forces measured between a calcium blocker drug and its receptor in living cells using atomic force microscope. AB - The adhesion force between the tip of an atomic force microscope cantilever derivatized with nimodipine (a calcium blocker, from the dihydropyridine class, currently used in clinical medicine for hypertension) and living cells of Saccharomyces cerevisiae (unicellular eukaryotes which portray ultrastructural features characteristic of higher eukaryotic cells) was measured. This methodology allowed us to locate (and visualize) pores on the cell surface which may be responsible for calcium transportation in the living cells. The interaction of the cantilever derivatized with the calcium blocker and a pore, which can be a calcium channel, is more intense than a non-derivatized cantilever and the pore. Outside the pore (on the rest of cell surface), a derivatized or a non-derivatized cantilever has the same pattern of adhesion force. The information obtained with this method is very important for the design of new, more potent and less toxic drugs for pharmacological use. PMID- 14527680 TI - How often does the myristoylated N-terminal latch of c-Abl come off? AB - The myristoylated N-terminal latching to the C-terminal lobe of c-Abl was recently demonstrated to be an important regulatory element for the kinase, playing a role similar to that of the tyrosine-phosphorylated C-terminal tail of c-Src. A potential mechanism for activating c-Abl is the dissociation of the myristoylated N-terminal latch. How often does this latch spontaneously come off? A recent theoretical model along with the experimental results of Superti-Furga, Kuriyan, and co-workers suggests that the equilibrium fraction of c-Abl in which the myristoylated N-terminal is unlatched is approximately 0.5%. PMID- 14527681 TI - Negatively charged residues adjacent to IFM motif in the DIII-DIV linker of hNa(V)1.4 differentially affect slow inactivation. AB - The effects on slow inactivation (SI) of charge substitutions, neutralizations, and reversals were studied for the negatively charged residues D1309 and EE1314,15 surrounding the IFM motif in the DIII-DIV cytoplasmic linker - the putative fast inactivation particle - of human skeletal muscle voltage-gated sodium channel (hNa(V)1.4). Changing aspartate (D) at position 1309 to glutamate (E) (substitution) did not strongly affect SI, whereas charge neutralization to glutamine (Q) and charge reversal to arginine (R) right-shifted the midpoint of the steady-state SI curve. Charge neutralization (D-->Q) at position 1309 also reduced the apparent valence associated with SI. Glutamates (E) at positions 1314 and 1315 were similarly mutated. Charge reversal (EE-->RR) right-shifted the steady-state SI curve and both reversal and substitution (EE-->DD) reduced its apparent valence. Charge neutralization (EE-->QQ) and reversal decreased the maximum probability of SI. These mutations also had differential effects on the rate of SI onset and recovery. These results suggest that charged residues in the DIII-DIV linker may interact with structures that control SI. PMID- 14527682 TI - A transient tobacco expression system coupled to MALDI-TOF-MS allows validation of the impact of differential targeting on structure and activity of a recombinant therapeutic glycoprotein produced in plants. AB - Tobacco-based transient expression was employed to elucidate the impact of differential targeting to subcellular compartments on activity and quality of gastric lipase as a model for the production of recombinant glycoproteins in plants. Overall N-linked glycan structures of recombinant lipase were analyzed and for the first time sugar structures of its four individual N-glycosylation sites were determined in situ by matrix-assisted laser desorption/ionization time of-flight mass spectrometry (MALDI-TOF-MS) on a trypsin digest without isolation or deglycosylation of the peptides. Three glycosylation sites contain both complex-type N-glycans and high-mannose-type structures, the fourth is exclusively linked to high-mannose glycans. Although the overall pattern of glycan structures is influenced by the targeting, our results show that the type of glycans found linked to a given Asn residue is largely influenced by the physico-chemical environment of the site. The transient tobacco system combined with MALDI-TOF-MS appears to be a useful tool for the evaluation of glycoprotein production in plants. PMID- 14527683 TI - Drug-sensitivity pattern analysis for study of functional relationship between gene products. AB - We have developed a method that we call 'drug-sensitivity pattern analysis', or DSPA, for analysis of protein function. Cells are transfected with cDNA of the test molecule, followed by analysis of the sensitivity of the transfected cells to multiple growth-inhibitory drugs. If two cDNA products have similar functions, their transfected cells should show similar drug-sensitivity patterns. The cDNAs of some signaling molecules were transfected into NIH3T3 or Ha-ras-transformed NIH3T3 (ras-NIH) cells and stable transfectants, which expressed high amounts of the gene product, were isolated. Chemosensitivity of the transfected clone was compared with the parental cells by the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide method using more than 40 drugs. The chemosensitivity changes caused by the transfected gene were calculated and expressed numerically as 'drug chemosensitivity index' (DCI). When the DCI values were analyzed by regression analysis, a significant positive relationship between IkappaBalpha superrepressor and dominant-negative IKKbeta and an inverse relationship between p53 and Mdm2 were consistent with previous reports. Thus, the DSPA method is useful for identifying functional similarities between gene products. PMID- 14527684 TI - Activation of CEA-CAM-1-mediated cell adhesion via CD98: involvement of PKCdelta. AB - CD98 is a multifunctional protein involved in amino acid transport and regulation of integrin-mediated cell adhesion. Herein, we demonstrated that CD98 stimulation by anti-CD98 antibodies induced CEA-CAM-1-mediated cell adhesion in BaF3 cells expressing CEA-CAM-1, and suggest that this might be responsible for compact clumping of F9 embryonic carcinoma cells by CD98 stimulation. CEA-CAM-1 was co immunoprecipitated by anti-CD98 antibody. CD98 stimulation induced the translocation of cytoplasmic protein kinase Cdelta (PKCdelta) to the cell adhesion sites, and rottlerin that inhibited the PKCdelta translocation abolished the cell aggregation without affecting integrin activation. The results suggested that CD98 stimulation could activate CEA-CAM-1-mediated cell adhesion independently of integrins. PMID- 14527685 TI - A prokaryotic alternative oxidase present in the bacterium Novosphingobium aromaticivorans. AB - The alternative oxidase (AOX) is a terminal oxidase present in the respiratory chain of all plants as well as some yeasts and trypanosomes, but has not previously been found in a prokaryote. We have identified an AOX homologue in Novosphingobium aromaticivorans, the first AOX found in a prokaryote. We have cloned the gene for the N. aromaticivorans AOX and showed it to have a terminal oxidase activity when heterologously expressed in Escherichia coli. We have also shown that this novel AOX is expressed in N. aromaticivorans cells, and that its expression level is greatly influenced by the oxygen level and carbon source of the growth media. PMID- 14527686 TI - Occurrence and neuroendocrine role of D-aspartic acid and N-methyl-D-aspartic acid in Ciona intestinalis. AB - Probes for the occurrence of endogenous D-aspartic acid (D-Asp) and N-methyl-D aspartic acid (NMDA) in the neural complex and gonads of a protochordate, the ascidian Ciona intestinalis, have confirmed the presence of these two excitatory amino acids and their involvement in hormonal activity. A hormonal pathway similar to that which occurs in vertebrates has been discovered. In the cerebral ganglion D-Asp is synthesized from L-Asp by an aspartate racemase. Then, D-Asp is transferred through the blood stream into the neural gland where it gives rise to NMDA by means of an NMDA synthase. NMDA, in turn, passes from the neuronal gland into the gonads where it induces the synthesis and release of a gonadotropin releasing hormone (GnRH). The GnRH in turn modulates the release and synthesis of testosterone and progesterone in the gonads, which are implicated in reproduction. PMID- 14527687 TI - Cationic lipids and cationic ligands induce DNA helix denaturation: detection of single stranded regions by KMnO4 probing. AB - Cationic lipids and cationic polymers are widely used in gene delivery. Using 1,2 dioleoyl-3-trimethylammonium-propane (DOTAP) as a cationic lipid, we have investigated the stability of the DNA in DOTAP:DNA complexes by probing with potassium permanganate (KMnO4). Interestingly, thymidines followed by a purine showed higher susceptibility to cationic ligand-mediated melting. Similar studies performed with other water-soluble cationic ligands such as polylysine, protamine sulfate and polyethyleneimine also demonstrated melting of the DNA but with variations. Small cations such as spermine and spermidine and a cationic detergent, cetyl trimethylammonium bromide, also rendered the DNA susceptible to modification by KMnO4. The data presented here provide direct proof for melting of DNA upon interaction with cationic lipids. Structural changes subsequent to binding of cationic lipids/ligands to DNA may lead to instability and formation of DNA bubbles in double-stranded DNA. PMID- 14527688 TI - High-throughput screening of structural proteomics targets using NMR. AB - We applied a high-throughput strategy for the screening of targets for structural proteomics of Xanthomonas axonopodis pv citri. This strategy is based on the rapid (1)H-(15)N HSQC NMR analysis of bacterial lysates containing selectively (15)N-labelled heterologous proteins. Our analysis permitted us to classify the 19 soluble candidates in terms of 'foldedness', that is, the extent to which they present a well-folded solution structure, as reflected by the quality of their NMR spectra. This classification allowed us to define a priority list to be used as a guide to select protein candidates for further structural studies. PMID- 14527689 TI - Adenovirus 2 E1B-55K protein relieves p53-mediated transcriptional repression of the survivin and MAP4 promoters. AB - It is well established that adenovirus E1B-55K protein functions as an inhibitor of the tumor suppressor protein p53 by binding and inactivating p53 as a transcriptional activator protein. Here we show that the adenovirus 2 E1B-55K protein also blocks p53 as a transcriptional repressor protein of the survivin and the MAP4 promoters. The repression is dependent on the ability of E1B-55K to bind to p53 and is enhanced by coexpression of the adenovirus E4orf6 protein. Overexpression of the transcriptional corepressor protein Sin3A partially relieves the inhibitory effect of E1B-55K, suggesting that E1B-55K blocks p53 functions by interfering with the Sin3 complex. PMID- 14527690 TI - Boric acid reversibly inhibits the second step of pre-mRNA splicing. AB - Several approaches have been used to identify the factors involved in mRNA splicing. None of them, however, comprises a straightforward reversible method for inhibiting the second step of splicing using an external reagent other than a chelator. This investigation demonstrates that the addition of boric acid to an in vitro pre-mRNA splicing reaction causes a dose-dependent reversible inhibition effect on the second step of splicing. The mechanism of action does not involve chelation of several metal ions; hindrance of 3' splice-site; or binding to hSlu7. This study presents a novel method for specific reversible inhibition of the second step of pre-mRNA splicing. PMID- 14527691 TI - Effective detection of remote homologues by searching in sequence dataset of a protein domain fold. AB - Profile matching methods are commonly used in searches in protein sequence databases to detect evolutionary relationships. We describe here a sensitive protocol, which detects remote similarities by searching in a specialized database of sequences belonging to a fold. We have assessed this protocol by exploring the relationships we detect among sequences known to belong to specific folds. We find that searches within sequences adopting a fold are more effective in detecting remote similarities and evolutionary connections than searches in a database of all sequences. We also discuss the implications of using this strategy to link sequence and structure space. PMID- 14527692 TI - Computational analysis of hydrogen bonds in protein-RNA complexes for interaction patterns. AB - Structural analysis of protein-RNA complexes is labor-intensive, yet provides insight into the interaction patterns between a protein and RNA. As the number of protein-RNA complex structures reported has increased substantially in the last few years, a systematic method is required for automatically identifying interaction patterns. This paper presents a computational analysis of the hydrogen bonds in the most representative set of protein-RNA complexes. The analysis revealed several interesting interaction patterns. (1) While residues in the beta-sheets favored unpaired nucleotides, residues in the helices showed no preference and residues in turns favored paired nucleotides. (2) The backbone hydrogen bonds were more dominant than the base hydrogen bonds in the paired nucleotides, but the reverse was observed in the unpaired nucleotides. (3) The protein-RNA complexes contained more paired nucleotides than unpaired nucleotides, but the unpaired nucleotides were observed more frequently interacting with the proteins. And (4) Arg-U, Thr-A, Lys-A, and Asn-U were the most frequently observed pairs. The interaction patterns discovered from the analysis will provide us with useful information in predicting the structure of the RNA binding protein and the structure of the protein binding RNA. PMID- 14527693 TI - Differential localization of lipid phosphate phosphatases 1 and 3 to cell surface subdomains in polarized MDCK cells. AB - Lipid phosphate phosphatases (LPPs) are integral membrane proteins with six transmembrane domains that act as ecto-enzymes dephosphorylating a variety of extracellular lipid phosphates. Using polarized MDCK cells stably expressing human LPP1 and LPP3, we found that LPP1 was located exclusively at the apical surface whereas LPP3 was distributed mostly in the basolateral subdomain. We identified a novel apical sorting signal at the N-terminus of LPP1 composed of F(2)DKTRL(7). In the case of LPP3, a dityrosine motif present in the second cytoplasmic portion was identified as basolateral targeting signal. Our work shows that LPP1 and LPP3 are equipped with distinct sorting signals that cause them to differentially localize to the apical vs. the basolateral subdomain, respectively. PMID- 14527694 TI - Enhanced gene silencing by the application of multiple specific small interfering RNAs. AB - Small interfering RNA duplexes (siRNA) induce gene silencing in various eukaryotic cells, although usually in an incomplete manner. Using chemically synthesized siRNAs targeting the HIV-1 co-receptor CXCR4 or the apoptosis inducing Fas-ligand (FasL), co-transfection of cells with two or more siRNA duplexes targeting different sites on the same mRNA resulted in an enhanced gene silencing compared with each single siRNA. This was shown in the down-regulation of protein and mRNA expression, and functionally in the inhibition of CXCR4 mediated HIV infection and of FasL-mediated cell apoptosis. Transfection efficiency determined for the FasL-specific siRNAs was dose-dependent and varied among the siRNAs tested, but was not the main reason for the enhanced gene silencing. PMID- 14527695 TI - Acyl-coenzyme A organizes laterally in membranes and is recognized specifically by acyl-coenzyme A binding protein. AB - Long chain acyl-coenzyme A (acyl-CoA) is a biochemically important amphiphilic molecule that is known to partition strongly into membranes by insertion of the acyl chain. At present, microscopically resolved evidence is lacking on how acyl CoA influences and organizes laterally in membranes. By atomic force microscopy (AFM) imaging of membranes exposed to acyl-CoA in microM concentrations, it is shown that aggregate formation takes place within the membrane upon long-time exposure. It is known that acyl-CoA is bound by acyl-CoA binding protein (ACBP) with high affinity and specificity and that ACBP may bind and desorb membrane bound acyl-CoA via a partly unknown mechanism. Following incubation with acyl CoA, it is shown that ACBP is able to reverse the formation of acyl-CoA aggregates and to associate peripherally with acyl-CoA on the membrane surface. Our microscopic results point to the role of ACBP as an intermembrane transporter of acyl-CoA and demonstrate the ability of AFM to reveal the remodelling of membranes by surfactants and proteins. PMID- 14527696 TI - Molecular assembly and subcellular distribution of ATP-sensitive potassium channel proteins in rat hearts. AB - Cardiac ATP-sensitive K(+) (K(ATP)) channels are proposed to contribute to cardio protection and ischemic preconditioning. Although mRNAs for all subunits of K(ATP) channels (Kir6.0 and sulfonylurea receptors SURs) were detected in hearts, subcellular localization of their proteins and the subunit combination are not well elucidated. We address these questions in rat hearts, using anti-peptide antibodies raised against each subunit. By immunoblot analysis, all of the subunits were detected in microsomal fractions including sarcolemmal membranes, while they were not detected in mitochondrial fractions at all. Immunoprecipitation and sucrose gradient sedimentation of the digitonin solubilized microsomes indicated that Kir6.2 exclusively assembled with SUR2A. The molecular mass of the Kir6.2-SUR2A complex estimated by sucrose sedimentation was 1150 kDa, significantly larger than the calculated value for (Kir6.2)(4) (SUR2A)(4), suggesting a potential formation of micellar complex with digitonin but no indication of hybrid channel formation under the conditions. These findings provide additional information on the structural and functional relationships of cardiac K(ATP) channel proteins involving subcellular localization and roles for cardioprotection and ischemic preconditioning. PMID- 14527698 TI - Hyperalgesia versus response bias in fibromyalgia. PMID- 14527699 TI - Causes of pain in degenerative bone and joint disease: a lesson from vertebroplasty. AB - Pain in degenerative bone and joint disease is usually attributed to sensitized nociceptors in inflamed periarticular soft tissues. Here we draw attention to the potential contribution of intrinsic bone innervation. The structure and innervation of articular bone ends is analogous to that of teeth. Although some dental pain derives from inflamed periodontal soft tissue, a more important source is the dentine and root canal. By analogy, pain on weight bearing in osteoarthritis and related conditions may be due to compressive forces applied to the innervation of subchondral bone exposed by erosion of the overlying cartilage. Pain relief obtained by injecting acrylic cement into the bone interior during percutaneous vertebroplasty is consistent with this concept. The development of a new family of pain relief options based on "marrow canal treatment" may be a realistic possibility. PMID- 14527700 TI - Silver nitrate cauterization: characterization of a new model of corneal inflammation and hyperalgesia in rat. AB - Chemical cauterization of the central cornea with silver nitrate was assessed as a superficial injury model of tissue sensitization accompanying acute inflammation. Adult male Sprague-Dawley rats were anesthetized with halothane gas, and the centers of their right corneas treated with a silver nitrate applicator stick (75% silver nitrate, 25% potassium nitrate) to produce a discrete lesion 1 mm in diameter. Edema of the corneal stroma and elevated immune cell counts became significant 4 h after cauterization, and were still evident after 48 h. Behavioral sensitization to chemical stimuli was determined by counting the number of blinks following application of 1 microM capsaicin directly to the corneal surface. A significant increase in stimulus-induced blinking was evident 2 h after cauterization. Chemical sensitization peaked at 6 h, and was no longer significant at 12 h. We conclude that silver nitrate cauterization produces acute corneal inflammation and hyperalgesia, and may prove a useful model for the study of primary afferent nociceptors. PMID- 14527701 TI - Increased pain sensitivity in fibromyalgia: effects of stimulus type and mode of presentation. AB - Fibromyalgia (FM) is defined in part by sensitivity to blunt pressure. Pressure pain sensitivity in FM is evaluated typically by the use of 'ascending' testing methods such as tender point counts or dolorimetry, which can be influenced by response bias of both the subject and examiner. Methods that present stimuli in a random, unpredictable fashion might minimize the influence of these factors. In this study, we compared the results of ascending and random assessments of both pressure and thermal pain sensitivities in 43 FM patients and 28 age- and gender matched controls. Even though FM is defined on the basis of pressure sensitivity, this group was also more sensitive to heat stimuli, presented in either ascending or random paradigms. In both the patient and control groups, the pain ratings to painful sensations evoked by both thermal and pressure stimuli were significantly greater in the random, compared with the ascending method. The number of subjects classified as 'expectant' because they rated pain higher in ascending than random paradigms was similar for FM and control groups. Both patients and controls exhibited a similar degree of sensitization to pressure and thermal stimuli. The increased sensitivity to both pressure and thermal stimuli for threshold and suprathreshold stimuli in FM patients is consistent with central augmentation of pain processing. PMID- 14527702 TI - Preoperative prediction of severe postoperative pain. AB - We developed and validated a prediction rule for the occurrence of early postoperative severe pain in surgical inpatients, using predictors that can be easily documented in a preoperative setting. A cohort of surgical inpatients (n=1416) undergoing various procedures except cardiac surgery and intracranial neurosurgery in a University Hospital were studied. Preoperatively the following predictors were collected: age, gender, type of scheduled surgery, expected incision size, blood pressure, heart rate, Quetelet index, the presence and severity of preoperative pain, health-related quality of life the (SF-36), Spielberger's State-Trait Anxiety Inventory (STAI) and the Amsterdam Preoperative Anxiety and Information Scale (APAIS). The outcome was the presence of severe postoperative pain (defined as Numeric Rating Scale > or =8) within the first hour postoperatively. Multivariate logistic regression in combination with bootstrapping techniques (as a method for internal validation) was used to derive a stable prediction model. Independent predictors of severe postoperative pain were younger age, female gender, level of preoperative pain, incision size and type of surgery. The area under the receiver operator characteristic (ROC) curve was 0.71 (95% CI: 0.68-0.74). Adding APAIS scores (measures of preoperative anxiety and need for information), but not STAI, provided a slightly better model (ROC area 0.73). The reliability of this extended model was good (Hosmer and Lemeshow test p-value 0.78). We have demonstrated that severe postoperative pain early after awakening from general anesthesia can be predicted with a scoring rule, using a small set of variables that can be easily obtained from all patients at the preoperative visit. Before this internally validated preoperative prediction rule can be applied in clinical practice to support anticipatory pain management, external validation in other clinical settings is necessary. PMID- 14527703 TI - Effects of local pressure and vibration on muscle pain from eccentric exercise and hypertonic saline. AB - In human subjects the triceps surae of one leg was exercised eccentrically by asking subjects to walk backwards on an inclined treadmill. Before the exercise controlled local pressure, applied to the muscle with an electromagnet, produced mild soreness, which was reduced when the pressure was combined with vibration. When delayed-onset muscle soreness (DOMS) had set in, 24-48 h after the exercise, vibration increased pain from local pressure. Vibrating at different frequencies suggested 80 Hz as the optimal frequency. During 2-h testing post-exercise, evidence of a change in character of the effects of vibration was first detected at 6 h. It persisted up to 72 h post-exercise. When muscle pain was generated in an unexercised triceps by injection of hypertonic (5%) saline, controlled local pressure applied to the sore area increased pain levels by 32% while pressure plus vibration reduced this to 11%. In a subject with DOMS, local pressure again increased pain from saline by 32% but combining it with vibration increased pain further by an additional 20%. The effect of vibration on DOMS could be abolished with a large nerve fibre block applied to the sciatic nerve. It is concluded that the vibration effects are the result of stimulation of large-diameter mechanoreceptive afferents in the muscle which, it is speculated, play a role in generating DOMS. PMID- 14527704 TI - The parents' postoperative pain measure: replication and extension to 2-6-year old children. AB - Pain assessment is a difficult task for parents at home following children's surgery. The purpose of the present study was to confirm the psychometric properties of a behavioural measure of postoperative pain developed to assist parents with pain assessment in children aged 7-12 years following day surgery. The study also examined the reliability and validity of the measure with children aged 2-6 years. Participants were 51 parents of children aged 7-12 years and 107 parents of children aged 2-6 years. For the 2 days following surgery, parents completed a pain diary that included global ratings of their children's pain and the 15-item Parents' Postoperative Pain Measure (PPPM). The older children provided self-reports of their pain intensity. The PPPM items showed good internal consistency on the two postoperative days for both samples (alpha's=0.81 0.88) and scores on the PPPM were highly correlated with children's (for the older children) and parents' (for the young children) global ratings of pain (r's=0.53-0.72). As global pain ratings decreased from Days 1 to 2, so did scores on the PPPM. Scores on the PPPM were successful in discriminating between children who had undergone low/moderate and high pain surgeries. The results of this study provide evidence of the reliability and validity of the PPPM as a measure of postoperative pain among children aged 2 through to 12 years. PMID- 14527705 TI - BOLD effects in different areas of the cerebral cortex during painful mechanical stimulation. AB - The contribution of four cortical areas (S1, S2, insular cortex and gyrus cinguli) to pain processing was assessed by functional magnetic resonance imaging (fMRI). Phasic (mechanical impact) and tonic stimuli (squeezing) were applied to the back of a finger, both at two different strengths. Stimuli were adjusted to inflict weak and strong pain sensations. It had been shown before that stronger noxious mechanical stimuli induce a weaker input from myelinated mechanoreceptors, but a more vigorous input from nociceptive primary afferents, and vice versa. Sizes of activation clusters and percent increase of the blood oxygenation level dependent (BOLD) signal during activation were compared in the areas of interest. Phasic stimulus patterns were more closely reflected in the time course of the MR signal in S1, S2 and the cingulate than tonic patterns, since the tonic stimuli tended to induce slow MR signal increase also during the resting periods which is in parallel to the persisting character of the tonic pain sensations. In S1 only the contralateral side was activated in most cases, and the more painful stimuli did not induce greater BOLD responses compared to the less painful stimuli in this area. Paradoxically, more painful stimuli produced smaller activation clusters in S1, both in tonic and phasic stimulus trials. In contralateral S2 more painful phasic stimuli induced significantly stronger BOLD responses than the weaker stimuli. The responses to tonic stimuli did not differentiate painfulness and were significantly smaller than the phasic. Activation clusters in this area were also smaller for tonic stimuli. In the gyrus cinguli more painful phasic stimuli induced stronger BOLD responses, but no difference was seen between tonic stimulation of different strength. Though the insular cortex was often bilaterally activated, no significant differences between stimulus quality or intensity were found. Our results provide evidence for a contribution of the S2 projection area and of the cingulate cortex to the processing of the intensity dimension of phasic mechanical pain. Such evidence was not found for the S1 area, which probably receives dominant input from non nociceptive mechanoreceptors. PMID- 14527706 TI - Effect of morphine sulphate eye drops on hyperalgesia in the rat cornea. AB - In addition to their traditional role in centrally mediated analgesia, opiate compounds produce significant effects when administered peripherally. Using a recently characterized model of acute chemical injury to the rat cornea, we assessed the effects of morphine sulphate eye drops on corneal inflammation and hyperalgesia. Topical application of a 5 microM morphine sulphate eye drop preparation attenuated capsaicin-induced blinking in a concentration-dependent manner. However, morphine had no effect on capsaicin-induced blinking when applied to healthy, non-inflamed rat cornea. In addition, 5 microM morphine given every 2 h following cauterization retarded the development of both stromal edema and the infiltration of immune cells. Both the analgesic and anti-inflammatory effects of morphine were prevented by the opioid receptor antagonists naloxone, CTAP, and naltrindole. We conclude that morphine acts on mu and delta opioid receptors located in the rat cornea to attenuate inflammation and hyperalgesia. PMID- 14527707 TI - Inflammation and hyperalgesia induced by nerve injury in the rat: a key role of mast cells. AB - Inflammatory cells and their mediators are known to contribute to neuropathic pain following nerve injury. Mast cells play a key role in non-neural models of inflammation and we propose that mast cells and their mediators (in particular histamine) are important in the development of neuropathic pain. In rats, where the sciatic nerve was partially ligated, we showed that stabilisation of mast cells with sodium cromoglycate reduced the recruitment of neutrophils and monocytes to the injured nerve and suppressed the development of hyperalgesia. Treatment with histamine receptor antagonists suppressed the development of hyperalgesia following nerve injury and alleviated hyperalgesia once it was established. These results suggest that mast cell mediators such as histamine released within hours of nerve injury contribute to the recruitment of leukocytes and the development of hyperalgesia. PMID- 14527708 TI - Analgesic and placebo effects of thalamic stimulation. AB - Numerous clinical studies have reported successful relief of chronic pain with sensory thalamic stimulation. However, even with the extensive use of sensory thalamic stimulation as a clinical tool in the relief of chronic pain, the results are still inconsistent. This discrepancy could probably be explained by the fact that the majority of these studies are case reports or retrospective analyses, which have often used imprecise pain measurements that do not allow a rigorous statistical evaluation of pain relief. None of these studies measured the effect of stimulation on clinical pain for longer than a few hours per day, which is an important aspect considering that clinical pain can vary over time. Moreover, placebo controls are seldom included. In the current study, we measured patients' pain perception at home over a 2-week period, both during days of normal stimulation of the sensory thalamus and during days without stimulation. Patients also came to the laboratory to assess the effects of thalamic and placebo stimulation on clinical pain, experimental heat pain, innocuous air puff and visual stimulation. A potential relation between the perceived paresthesia and analgesic efficacy during thalamic and placebo stimulation was also explored. We found that thalamic stimulation significantly affected clinical and experimental pain perception, but that an important placebo component also exists. On the other hand, neither thalamic nor placebo stimulation affected air puff and visual ratings, suggesting that the effect applies specifically to pain and hence is not caused by a general change in attention. The level of paresthesia elicited during the placebo manipulation was also directly correlated with the degree of placebo pain relief. These results suggest that thalamic stimulation produces a small but significant reduction in pain perception, but that a significant placebo effect also exists. PMID- 14527709 TI - Role of nerve growth factor in the trinitrobenzene sulfonic acid-induced colonic hypersensitivity. AB - The majority of patients with digestive disorders display visceral pain. In these troubles, visceral pain threshold is decreased, demonstrating visceral hypersensitivity. There is growing evidence that nerve growth factor (NGF) may function as a mediator of persistent pain states. This hypothesis was tested in a model of colonic hypersensitivity measured by isobaric distension in conscious rats. This study was designed to evaluate (1) the effect of exogenous NGF on colonic pain threshold, (2) the involvement of NGF in trinitrobenzene sulfonic acid (TNBS)-induced colonic hypersensitivity, by testing an anti-NGF antibody, and (3) finally the involvement of sensory nerves on NGF and TNBS effects using rats treated neonatally with capsaicin. Intra-peritoneal injection of NGF (0.1 100 ng/rat) decreased in a dose-related manner colonic pain threshold in naive rats. This effect was reversed by anti-NGF antibody (1/2000; 2 ml/kg). TNBS induced colonic hypersensitivity was also reversed by anti-NGF antibody (1/2000; 2 ml/kg): 37.7 +/- 1.7 and 17.6 +/- 0.7 mmHg (p<0.01) for anti-NGF antibody- and vehicle-treated group, respectively. Neonatal capsaicin pre-treatment inhibited NGF- and TNBS-induced decrease in colonic pain threshold: 49.4 +/- 5.3 versus 22.3 +/- 1.6 mmHg (p<0.01) for capsaicin versus vehicle in NGF-treated rats and 39.6 +/- 3.3 versus 18.0 +/- 1.0 mm Hg (p<0.001) for capsaicin versus vehicle in TNBS-treated rats. These data suggest that the action of NGF on sensory neurons contributes to the development of visceral hypersensitivity and that anti-NGF strategy may be of some therapeutic benefits in digestive sensory disorders. PMID- 14527710 TI - QTc interval prolongation associated with intravenous methadone. AB - Numerous medications prolong the rate-corrected QT (QTc) interval and induce arrhythmias by blocking ionic current through cardiac potassium channels composed of subunits expressed by the human ether-a-go-go-related gene (HERG). Recent reports suggest that high doses of methadone cause torsades de pointes. To date, no controlled study has described an association between methadone and QTc prolongation. The only commercial formulation of parenteral methadone available in the United States contains the preservative chlorobutanol. The objectives of this study are to determine: (1) whether the administration of intravenous (i.v.) methadone causes QTc prolongation in humans; (2) whether methadone and/or chlorobutanol block cardiac HERG potassium currents (IHERG) in vitro. Over 20 months, we identified every inpatient with at least one electrocardiogram (ECG) performed on i.v. methadone. For each patient, we measured QTc intervals for every available ECG performed on and off i.v. methadone. Concurrent methadone doses were also recorded. Similar data were collected for a separate group of inpatients treated with i.v. morphine. In a separate set of experiments IHERG was evaluated in transfected human embryonic kidney cells exposed to increasing concentrations of methadone, chlorobutanol, and the two in combination. Mean difference (+/- standard error) per patient in QTc intervals on and off methadone was 41.7 (+/- 7.8)ms, p<0.0001. Mean difference in QTc intervals on and off morphine was 9.0 (+/- 6.1)ms, p=0.15. The approximately linear relationship between QTc measurements and log-dose of methadone was significant (p<0.0001). Methadone and chlorobutanol independently block IHERG in a concentration dependent manner with IC50 values of 20 +/- 2 microM and 4.4 +/- 0.3 mM, respectively. Chlorobutanol potentiates methadone's ability to block IHERG. Methadone in combination with chlorobutanol is associated with QTc interval prolongation. Our data strongly suggest that methadone in combination with chlorobutanol is associated with QTc interval prolongation. PMID- 14527711 TI - Comment on Sterling, M., et al., Motor system dysfunction following whiplash injury, Pain 103 (2003) 65-73. PMID- 14527713 TI - A POLYCOMB group gene of rice (Oryza sativa L. subspecies indica), OsiEZ1, codes for a nuclear-localized protein expressed preferentially in young seedlings and during reproductive development. AB - The SET domains are conserved amino acid sequences present in chromosomal proteins that contribute to the epigenetic control of gene expression by altering regional organization of the chromatin structure. The SET domain proteins are divided into four subgroups as categorized by their Drosophila members; enhancer of zeste (E(Z)), trithorax (TRX), absent small or homeotic 1 (ASH1) and supressor of variegation (SU(VAR)3-9). Homologs of all four classes have been characterized in yeast, mammals and plants. We report here the isolation and characterization of rice (Oryza sativa L. subspecies indica) cDNA, OsiEZ1, as a monocot member of this family. The OsiEZ1 cDNA is 3133 bp long with an ORF of 2799 bp, and the predicted amino acid sequence (895 residues) corresponds to a protein of ca. 98 kDa. All the characteristic domains known to be conserved in E(Z) homologs (subgroup I) of SET domain containing proteins are present in OsiEZ1. In the rice genome, a 7499 bp long OsiEZ1 sequence is split into 17 exons interrupted by 16 introns. Southern analysis indicates that OsiEZ1 is represented as single copy in the rice genome. Expression studies revealed that the OsiEZ1 transcript level was highest in rice flowers, almost undetectable in developing seeds of 1-2 days post fertilization but increased significantly in young seeds of 3-5 days post fertilization. The OsiEZ1 transcript was barely detectable in mature zygotic embryos, but its levels were significantly higher in callus derived from rice scutellum, somatic embryos and young seedlings. The OsiEZ1/GUS recombinant protein was confined to the nucleus in living cells of particle-bombarded onion peels. The expression of OsiEZ1 complemented a set1Delta Saccharomyces cerevisiae mutant that is impaired in telomeric silencing. We suggest that the nuclear localized OsiEZ1 has a role in regulating various aspects of plant development, and this control is most likely brought about by repressing the activity of downstream regulatory genes. PMID- 14527714 TI - Identification and characterization of a new human type 9 cGMP-specific phosphodiesterase splice variant (PDE9A5). Differential tissue distribution and subcellular localization of PDE9A variants. AB - Previously, four splice variants of human cGMP-specific phosphodiesterase (PDE) 9A (PDEs 9A1, 9A2, 9A3 and 9A4) have been identified. In this study, we have cloned a cDNA representing a new human PDE9A variant (PDE9A5). PDE9A5 encodes a protein of 492 amino acids, smaller than PDEs 9A1 and 9A2 but larger than PDEs 9A3 and 9A4. The exon structure of PDE9A5 is different from those of PDEs 9A1, 9A2, 9A3 and 9A4 in that, of the 20 exons of PDE9A gene, it lacks exons 2 and 5. PDE9A5 has been characterized in comparison with PDE9A1, the longest PDE9A variant. PDEs 9A5 and 9A1 have similar enzymatic properties. They both have a high affinity for cGMP with similar Km values (0.39 and 0.25 microM, respectively), although they have slightly different Vmax values (2.55 and 0.96 micromol/min/mg, respectively). They exhibit very similar divalent metal ion dependency and inhibitor sensitivity. Real-time quantitative PCR analysis shows that PDEs 9A5 and 9A1 exhibit differential tissue distribution. They are highly expressed in immune tissues (spleen, lymph node and thymus) and are more abundant in T cells than in B cells, neutrophils and monocytes. When transiently expressed in HEK293 cells, PDEs 9A5 and 9A1 proteins exhibit differential subcellular localization. PDE9A5 localizes exclusively in the cytoplasm, whereas PDE9A1 localizes in the nucleus only. The nuclear localization of PDE9A1 is dependent on a unique pat7 motif. By Western blot analysis, native PDE9A1 is detectable in the nucleus but not in the cytoplasm of T cells. Thus, to our knowledge, PDE9A1 is the only PDE isoform found to localize exclusively in the nucleus. We speculate that the physiological role of the PDE9A diversity may be imparting cGMP metabolizing ability to specific cellular compartments in appropriate tissues. PMID- 14527716 TI - A novel gene encoding a coiled-coil mitochondrial protein located at the telomeric end of the human MHC Class III region. AB - The human Major Histocompatibility Complex (MHC) Class III region, which lies in between the MHC Class I and Class II regions on chromosome 6p21.3, contains approximately 60 genes with diverse functions. Using bioinformatics analyses, we identified a novel open reading frame (ORF) in this region, telomeric of BAT1, which we called Mitochondrial Coiled-Coil Domain 1 (MCCD1). The expression of the predicted ORF in a number of human tissues was confirmed by RT-PCR analysis. An orthologue of the MCCD1 gene was identified in the swine MHC in an analogous position, adjacent to pig BAT1. The overall sequence identity between the human and pig MCCD1 proteins is only 65.9%, but their C-terminal domains are highly conserved, showing 92% identity over 53 residues. The MCCD1 gene encodes a short polypeptide (119 amino acids) which contains a putative coiled-coil domain at its highly conserved C terminus and a predicted mitochondrial localisation signal at its N terminus. Transient expression in mammalian cells of MCCD1 fused at its C terminus to either EGFP or the T7-epitope tag showed that this protein is indeed targeted to mitochondria. Finally, we characterised the polymorphism in this gene using denaturing high-performance liquid chromatography (DHPLC) analysis and found that the MCCD1 gene is highly polymorphic containing an average of 1 single nucleotide polymorphism (SNP) every 99 bp. Interestingly, MCCD1 contains four SNPs within the coding region, three of which cause nonsynonymous and nonconservative changes in the amino acid sequence. PMID- 14527719 TI - Genomic DNA analysis of genes encoding (hemi-)cellulolytic enzymes of the anaerobic fungus Piromyces sp. E2. AB - Anaerobic fungi contain more than one copy of genes encoding (hemi-)cellulases in their genome. The arrangement of these genes on the chromosomes was not known. A genomic DNA (gDNA) library of Piromyces sp. E2 was screened with different probes specific for (hemi-)cellulolytic enzymes. This screening resulted in three gDNA clones with genes encoding glycoside hydrolase enzymes of families 1 (beta glucosidase), 6 (exoglucanase) and 26 (mannanase). Each clone contained two or more genes of the same family. Comparison of the gene copies on a clone revealed that they were highly homologous, and in addition, 54-75% of the substitutions was synonymous. One of the mannanase genes contained an intron. PCR with selected primers resulted in a gDNA clone with a new representative (cel9B) of glycoside hydrolase family 9 (endoglucanase). Comparison with cel9A revealed that cel9B had 67% homology on the nucleotide level. Furthermore, three introns were present. All results of this paper taken together provided evidence for duplications of (hemi-)cellulolytic genes, which resulted in clusters of almost identical genes arranged head-to-tail on the genome. In contrast to other eukaryotes, this phenomenon appears frequently in anaerobic fungi. PMID- 14527717 TI - Identification of the Drosophila progenitor of mammalian Kruppel-like factors 6 and 7 and a determinant of fly development. AB - The Kruppel-like transcription factors (KLFs) represent a family of 15 different zinc finger proteins of the C(2)H(2) type that are involved in vertebrate development and which control cell proliferation, growth and differentiation. Structural-functional considerations have segregated KLF6 and KLF7 into a phylogenetically distinct group. Here we report the identification of Luna, the Drosophila progenitor of the mammalian KLF6/KLF7 group. This conclusion is based on the near sequence identity, as well as the comparable location of the DNA binding domains and nuclear localization signals of the insect and mammalian proteins. The homology extends to the composition and function of the amino terminal segment of Luna which, similarly to the mammalian counterparts, stimulates transcription in a reporter gene assay. We also present preliminary in vivo evidence of Luna involvement in embryonic development and cell differentiation. First, luna RNA interference and luna overexpression during early Drosophila embryogenesis leads to developmental arrest at different embryonic stages. Second, targeted perturbation of luna expression in the forming compound eye interferes with terminal cell differentiation, but not cell specification. We therefore propose that Luna is a novel transcriptional determinant of Drosophila development. PMID- 14527718 TI - Functional characterization of two flap endonuclease-1 homologues in rice. AB - Flap endonuclease-1 (FEN-1) is an important enzyme involved in DNA replication and repair. Previously, we isolated and characterized a complementary DNA (cDNA) from rice (Oryza sativa) encoding a protein which shows homology with the eukaryotic flap endonuclease-1 (FEN-1). In this report, we found that rice (O. sativa L. cv. Nipponbare) possessed two FEN-1 homologues designated as OsFEN-1a and OsFEN-1b. The OsFEN-1a and OsFEN-1b genes were mapped to chromosome 5 and 3, respectively. Both genes contained 17 exons and 16 introns. Alignment of OsFEN-1a protein with OsFEN-1b protein showed a high degree of sequence similarity, particularly around the N and I domains. Northern hybridization and in situ hybridization analysis demonstrated preferential expression of OsFEN-1a and OsFEN 1b in proliferating tissues such as the shoot apical meristem or young leaves. The levels of OsFEN-1a and OsFEN-1b expression were significantly reduced when cell proliferation was temporarily halted by the removal of sucrose from the growth medium. When the growth-halted cells began to regrow following the addition of sucrose to the medium, both OsFEN-1a and OsFEN-1b were again expressed at high level. These results suggested that OsFEN-1a and OsFEN-1b are required for cell proliferation. Functional complementation assay suggested that OsFEN-1a cDNA had the ability to complement Saccharomyces cerevisiae rad27 null mutant. On the other hand, OsFEN-1b cDNA could not complement the rad27 mutant. The roles of OsFEN-1a and OsFEN-1b in plant DNA replication and repair are discussed. PMID- 14527720 TI - The human MJD gene: genomic structure and functional characterization of the promoter region. AB - Machado-Joseph disease (MJD) is a progressive neurodegenerative disorder caused by expansion of a CAG motif within the translated region of the human MJD (hMJD) gene which has been mapped to chromosome 14q. In this study, the hMJD gene was identified in two overlapping bacterial artificial chromosome (BAC) clones and contained 11 exons resulting in a 6.14 kb transcript. The 5'-flanking region of the hMJD gene included a TATA-less promoter with GC-rich regions, a CCAAT box and multiple potential SP1 binding sites. Luciferase reporter assays performed in neuronal and non-neuronal human cell lines demonstrated a core promoter within the 200 bp region immediately upstream of the putative transcriptional start site (-89 according to the start codon). DNA-protein interactions defined by electrophoretic mobility shift assays (EMSA) revealed specific binding of nuclear proteins to the putative core promoter region. PMID- 14527715 TI - Towards a catalogue of genes transcribed in multiple tissues of Mytilus galloprovincialis. AB - Mussels are among the most studied marine organisms because of their ecological role, economic importance and long-standing use in coastal pollution biomonitoring. Despite the bulk of data derived from traditional investigation areas, only limited knowledge is available on mussel genes and their expression in ordinary or stressing conditions. We present here the first data obtained by production and sequencing of 3' end-specific expressed sequence tags (3'ESTs) from multiple tissues of putatively unstressed mussels (Mytilus galloprovincialis). A total of 524 clusters (98 virtual consensuses and 426 singletons) derived from 829 reliable 3'ESTs were searched in the non-redundant (nr) sequence databases (National Centre for Biotechnology Information (Bethesda, USA), NCBI). Most contigs (59%) showed poor or no similarity, thus revealing unknown mussel genes, other contigs recognized some of the few available records of M. galloprovincialis (e.g. actin, cytochrome oxidase III and twitchin) and a number of significant similarities with mitochondrial (mt) or nuclear genes from other organisms were found. Actually, a variety of ribosomal as well as motility- and adhesion-related genes and some genes potentially involved in stress responses (e.g. myticin A, heat shock proteins, methallothionein) were putatively identified. Uniquely, mitochondrial transcripts were primarily represented by a cluster of 123 3'ESTs (1296 base pairs (bp) of the mussel 16S rRNA). In our knowledge, these results provide the first systematic production and annotation of ESTs in M. galloprovincialis. They also represent the first report of a wider project, based on the strategy of 3'EST identification from normal and stressed mussels and intended to define a mussel complementary DNA (cDNA) microarray for genome-wide transcription studies. PMID- 14527721 TI - Shadoo, a new protein highly conserved from fish to mammals and with similarity to prion protein. AB - We report evidence from cDNA isolation and expression analysis as well as analyses of genome, expressed sequence tag (EST), cDNA and expression databases for a new gene named SPRN (shadow of prion protein). SPRN comprises two exons, with the open reading frame (ORF) contained within exon 2, and codes for a protein of 130-150 amino acids named Shadoo (Japanese shadow), predicted to be extracellular and GPI-anchored. The SPRN gene was found in fish (zebrafish, Fugu) and mammals (mouse, rat, human). Conservation of order and transcription orientation of two proximal genes between fishes and mammals strongly indicates gene orthology. Sequence comparison shows: a highly conserved N-terminal signal sequence; Arg-rich basic region containing up to six tetrarepeats of consensus XXRG (where X is G, A or S); a hydrophobic region of 20 residues with strong homology to PrP; a less conserved C-terminal domain containing a conserved glycosylation motif; and a C-terminal peptide predicted to be a signal sequence for glycophosphotidylinositol (GPI)-anchor attachment. Fish Shadoos (Sho) show well conserved sequences (identity 54%) over 106 amino acids (zebrafish length), and conservation among the mammalian sequences is very high (identity 81-96%). The fish and mammalian sequences are also well conserved, particularly for zebrafish, to beyond the end of the hydrophobic sequence (identity 41-53%, 78 amino acids, zebrafish length). The overall structure appears closely related to prion proteins (PrPs), although the C-terminal domains of Shos are quite different from those of PrPs, for which conformational changes in mammals are implicated in disease. The structural similarity is particularly interesting given recent reports of three new genes with similarities to PrPs found in Fugu (PrP-like, PrP-461/stPrP-1 and stPrP-2) and other fish, but for which direct evolution to higher vertebrate PrPs is unlikely and for which no other mammalian homologues have been found. Database information indicates expression of SPRN in embryo, brain and retina of mouse and rat, hippocampus of human, and in embryo and retina of zebrafish, and we directly confirmed a strikingly specific expression of the mammalian (human, mouse, rat) transcripts in whole brain. This result together with some common structural features led to the suggestive hypothesis of a possible functional link between mammalian PrP and Sho proteins. PMID- 14527722 TI - Mapping and sequencing of acetylcholinesterase genes from the platyhelminth blood fluke Schistosoma. AB - Acetylcholinesterase (AChE) on the surface of the parasitic blood fluke Schistosoma is the likely target for schistosomicidal anticholinesterases. Determination of the molecular structure of this drug target is key for the development of improved anticholinesterase drugs and potentially a novel vaccine. We have recently cloned the cDNA encoding the AChE from the human parasite Schistosoma haematobium and succeeded in expressing functional recombinant protein. We now describe the cloning and molecular characterisation of homologues from two other schistosome species-Schistosoma mansoni and Schistosoma bovis, which are important parasites of man and cattle, respectively, but which differ in their sensitivity to the therapeutic anticholinesterase metrifonate. Comparison of the deduced amino acid sequences revealed that the AChE from all three species posses a high degree of identity, with conservation of all of the residues known to be important for substrate binding and catalytic activity. Also conserved is a unique C-terminal domain which is unusual in that it lacks the consensus for GPI modification, even though the native protein is considered to be GPI-anchored. We have also established the AChE gene structures for all three species and cloned the complete gene for S. haematobium AChE. The gene structure is relatively complex, comprising nine coding exons; the location of the splice sites is identical in all three species, but the size of the introns varies considerably. The two C-terminal splicing sites that are conserved in all species are also present in Schistosoma, but a third C-terminal conserved splicing site which is located 11-13 amino acids upstream of the histidine of the catalytic triad in all invertebrate AChE genes characterised to date is absent. We discuss our findings in the context of the molecular phylogeny of the AChE genes and the potential application to the control of schistosomiasis. PMID- 14527723 TI - Identification and characterization of C3orf6, a new conserved human gene mapping to chromosome 3q28. AB - This study reports the characterization of a novel human gene, chromosome 3 open reading frame 6 (C3orf6), mapped to chromosome 3q28, within the critical region of hereditary spastic paraplegia SPG14 locus. Based on computational "spliced" EST alignment and RT-PCR, two C3orf6 transcript variants were identified. The longer C3orf6 transcript contains a 1449-nt ORF, encoding a protein of 482 aa, while the shorter variant contains a 921-nt ORF, encoding for a protein of 306 aa. C3orf6 gene is organised on 12 exons and the shorter transcript comes from an alternative splicing event skipping exon 6. The two mRNA are differentially expressed in brain and in several other human tissues with a predominant level for the shorter transcript. By database analysis, EST assembling and RT-PCR, we identified the transcripts of mouse and rat C3orf6 orthologous genes. The involvement of C3orf6 in the spastic paraplegia was investigated by sequencing all coding exons and flanking sequences in the SPG14 family, excluding the presence of causative mutations. PMID- 14527724 TI - A novel third gonadotropin-releasing hormone receptor in the medaka Oryzias latipes: evolutionary and functional implications. AB - Gonadotropin-releasing hormone (GnRH) plays pivotal roles in the regulation of vertebrate reproduction through binding to its specific membrane receptor. Within the past few years, substantial evidence has accumulated that more than one GnRH receptor (GnRH-R) is expressed in individual vertebrate species. Two GnRH-Rs, termed GnRH-R1 and GnRH-R2, have been identified in a teleost, the medaka Oryzias latipes. Here we describe the identification and characterization of a novel third member of GnRH-R, designated GnRH-R3, in the medaka. GnRH-R3 share high sequence homology (77% amino acid identity in the transmembrane domain) with GnRH R1. Phylogenetic analysis and genetic mapping demonstrated that both GnRH-R1 and GnRH-R3 were orthologous to the type 2 GnRH-R in primates and that these two medaka receptors were duplicates resulting from the genome-wide duplication within the teleost lineage. GnRH-R3, however, contained three introns, whereas GnRH-R1 had only two. Moreover, unlike GnRH-R1, GnRH-R3 exhibited an approximately equal selectivity for two of three native GnRH forms in the medaka, chicken-II-type GnRH (cGnRH-II) and salmon-type GnRH (sGnRH), and a less sensitivity for the other form, medaka-type GnRH. GnRH-R3 was found to be expressed throughout the brain, and thus appeared to mediate the neuromodulatory functions of both cGnRH-II and sGnRH. These data identify GnRH-R3 as a new member of GnRH-R that arose in a recent genome duplication but has distinctive genomic structure and functional characteristic. PMID- 14527725 TI - The Fem1c genes: conserved members of the Fem1 gene family in vertebrates. AB - The fem-1 gene of Caenorhabditis elegans functions in a signaling pathway that controls sex determination. Homologs of fem-1 in mammals have been characterized, consisting of two family members, Fem1a and Fem1b. We report here on Fem1c, a third member of the Fem1 gene family, in three vertebrate species: human, mouse, and zebrafish. The proteins encoded by these Fem1c genes share >99% amino acid identity between human and mouse, 79% amino acid identity between mouse and zebrafish, and end with a C-terminal Arginine residue, which distinguishes them from other FEM-1 proteins reported thus far. The human and mouse Fem1c coding regions show conservation of intron-exon structure and expression pattern in adult tissues. Human FEM1C maps to 5q22, mouse Fem1c maps to chromosome 18, and zebrafish fem1c maps to Linkage Group 8. The Fem1c genes in vertebrates may play a conserved role in the development and/or physiologic function of these organisms. PMID- 14527726 TI - cDNA-AFLP analysis of inducible gene expression in rice seminal root tips under a water deficit. AB - The seminal roots of an upland rice variety, Azucena, showed accelerated elongation in response to a water deficit. The elongation of cortical cells in the elongation zone is rapidly stimulated within 16 h by the water deficit. cDNA AFLP analysis was used to examine gene expression in seminal root tips at four time points (4, 16, 48 and 72 h) during the water deficit. One hundred and six unique genes induced by the water deficit were obtained. The expression patterns of these genes were confirmed by Northern blot analysis based on 21 selected genes representing different patterns. The 106 upregulated genes were composed of 60 genes of known function, 28 genes of unknown function and 18 novel genes. Sixty genes of known functions were involved in transport facilitation, metabolism and energy, stress- and defense-related proteins, cellular organization and cell-wall biogenesis, signal transduction, expression regulator and transposable element, suggesting that seminal root tips undergo a complex adaptive process in response to the water deficit. Expression of 22 genes reached a maximum within 16 h of water deficit treatment. These included aquaporin (PIP2a), 9-cis-epoxycarotenoid dioxygenase (NCED1) and a negative regulator of gibberellin signal transduction (SPY); eight other genes participated in cell wall loosening or vesicle traffic. PMID- 14527727 TI - Distribution of the MCS4 RNA genes in mycoplasmas belonging to the Mycoplasma mycoides cluster. AB - MCS4 RNA is one of the small stable RNAs found in Mycoplasma capricolum subsp. capricolum type strain California kid. This RNA has a sequence similarity to that of eukaryotic U6 snRNA. There are two genes encoding MCS4 RNA, designated mcs4a and mcs4b, in the genome. Homologous sequences of these genes were not found in databases of other bacterial sequences. We searched for MCS4 RNA and its genes in other bacteria by PCR and hybridization techniques. The results strongly suggested that this RNA exists only in a limited species of mycoplasmas belonging to the Mycoplasma mycoides cluster. PMID- 14527729 TI - Differential expression of presenilin-alpha and -beta (PSalpha and PSbeta) in Xenopus laevis: embryonic phosphorylation of PSalpha. AB - Mutations in genes encoding the highly homologous proteins, presenilin-1 and -2 (PS1 and PS2), are linked to the development of early-onset Alzheimer's disease. On the other hand, presenilins are known to play a critical role(s) in cell fate decisions during embryonic development in Caenorhabditis elegans. The messenger RNAs (mRNAs) of amphibian presenilin homologues PSalpha and PSbeta are most abundantly synthesized in the brain and the ovary, but are differentially degraded upon oocyte maturation and at the midblastula transition (MBT), respectively. In this study, we examined the spatiotemporal distribution of PSalpha and PSbeta proteins and their post-translational modification. The results were essentially consistent with the mRNA data and revealed moreover that PSalpha was present exclusively as processed molecules in the early embryos, while PSbeta was present mainly as unprocessed molecules (90%). Furthermore, the C-terminal fragment (CTF) of PSalpha was phosphorylated upon oocyte maturation and dephosphorylated at MBT, while no phosphorylation of the PSbeta CTF was detectable. Human PS1 CTF exogenously injected was also phosphorylated in Xenopus oocytes induced to mature in vitro by progesterone treatment. Two phosphorylation loci were mapped at Thr(320) and Ser(334) in the hydrophilic loop region of PSalpha. Our results suggest that PS1 and PS2 may play different roles under physiological conditions despite their high structural similarity. PMID- 14527728 TI - Overexpression of CYP4G19 associated with a pyrethroid-resistant strain of the German cockroach, Blattella germanica (L.). AB - A new cytochrome P450 gene, CYP4G19, was identified and isolated as a differentially expressed gene between insecticide susceptible ACY and resistant Apyr-R German cockroach strains using PCR-selected subtractive hybridization and cDNA array techniques. The cDNA sequence of CYP4G19 has an open ready frame of 1638 nucleotides encoding a putative protein of 546 amino acid residues. Sequence analysis shows that CYP4G19 putative protein contains (1) a highly hydrophobic N terminus, (2) a P450 protein signature motif, FXXGXRXCXG, known to be an important ligand for heme binding, and (3) an important characteristic motif, EVDTFMFEGHDTT, for the family 4. Northern blot analysis indicated that levels of CYP4G19 expression were low in eggs, nymphs, and adults of the susceptible ACY strain with a similar expression pattern. The expression of CYP4G19 in the resistant Apyr-R strain was developmentally regulated, with very low expression in eggs, increasing in nymphs, and reaching a maximum in both female and male adults. Comparison of CYP4G19 expression between ACY and Apyr-R strains indicated that there was no difference in their eggs, but expression was higher ( approximately 1.7-fold) in the nymphs and much higher ( approximately 5-fold) in the male and female adults of the Apyr-R strain. Levels of CYP4G19 mRNA were readily detectable in head+thorax tissues and increased ( approximately 5-fold) in the abdomens of the ACY strain. In the Apyr-R strain, however, levels of CYP4G19 mRNA were relatively low in head+thorax tissues and were about 7-fold increase in the abdomen. Although expression patterns of CYP4G19 in head+thorax and abdomen tissues were similar (i.e. lower in head+thorax tissues and higher in abdomen tissues) in both the ACY and Apyr-R strains, the overexpression was more evident in the Apyr-R strain in both head+thorax and abdomen tissues than in the ACY strain. PMID- 14527730 TI - Protein BmP0 from the silkworm Bombyx mori can be assembled and is functional in the Saccharomyces cerevisiae ribosomal stalk in the absence of the acidic P1 and P2 proteins. AB - The DNA complementary to RNA (cDNA) of the ribosomal stalk protein BmP0 of the silkworm Bombyx mori was isolated from a cDNA library and was subsequently expressed in the conditional P0-null mutant Saccharomyces cerevisiae D67dGP0, whose ribosomes also lack the other stalk components, proteins P1/P2. The transformed strain was able to grow under restrictive conditions, indicating that in the absence of the P1/P2 proteins BmP0 can bind to the yeast ribosomes and complement the lack of the endogenous YP0 protein. In addition, the binding capacity of the B. mori ribosomal stalk components to the ribosomal particle was studied by means of high salt treatment of purified ribosomes. The BmP0 protein retained its binding to the ribosome, suggesting a stable association with the rRNA, in contrast to the acidic proteins BmP1 and BmP2, which were easily released. The results clearly indicate that, as opposed to recent in vitro results, BmP0 does not require the presence of P1/P2 proteins in order to bind to the ribosome. PMID- 14527731 TI - Human poly(ADP-ribose) glycohydrolase (PARG) gene and the common promoter sequence it shares with inner mitochondrial membrane translocase 23 (TIM23). AB - Poly(ADP-ribosyl)ation is a posttranslational protein modification mediated by members of the poly(ADP-ribose) polymerase (PARP) family. The ADP-ribose polymers, synthesized by the diverse PARP enzymes by cleavage of NAD(+), are involved in the regulation of multiple cellular functions. At present, only a single enzyme, poly (ADP-ribose) glycohydrolase (PARG), has been identified to catalyze ADP-ribose polymer hydrolysis in the cell causing a rapid turnover of the biopolymer which may ultimately result in lethal depletion of cellular NAD(+) pools. In this study, we describe the construction of the first human PARG cDNA clone by reverse transcription of CF3 human fibroblast RNA. Using the NCBI "Genome BLAST" program, the human PARG gene was mapped to chromosome 10 (10q11.23) in agreement to earlier results obtained by in situ hybridization. In vitro coupled transcription and translation of the cDNA yielded several specific bands in the range of 111-85 kDa, indicating possible usage of alternative translation initiation sites. The gene structure was characterized by further detailed computational analyses. The open reading frame consists of 18 exons and 17 introns with exons 9 to 14 forming the catalytic center of the enzyme and exons 1 to 3 encoding the putative regulatory domain. We show that the human PARG gene shares a 470-bp common promoter region with the inner mitochondrial membrane translocase 23 (TIM23). The human bidirectional promoter region was cloned and expression studies in transiently transfected HEK293 cells was performed using an EGFP-luciferase reporter fusion gene (GFL) to quantify transcription activation in both directions. The activity of the promoter was found to be 3.7 fold higher for TIM23 than for PARG, indicating that the two genes are expressed at different levels, although coregulation of the two genes remains an interesting possibility. PMID- 14527732 TI - Frameshift mutation events in beta-glucosidases. AB - Compensated frameshift mutation is a modification of the reading frame of a gene that takes place by way of various molecular events. It appears to be a widespread event that is only observed when homologous amino acid and nucleodotide sequences are compared. To identify these mutation events, the sequence analysis rationale was based on the search for short regions that would have much lower degrees of conservation in protein, but not in DNA, in well conserved beta-glucosidase families. We have restricted our study to a seed set of sequences of O-glycoside hydrolase families 1 and 3. We found compensated frameshift mutation in the family of 1 beta-glucosidases for the Erwinia herbicola, Cellulomonas fimi, and (non-cyanogenic) Trifolium repens gene sequences, and in the family of 3 beta-glucosidases for the Clostridium thermocellum and Clostridium stercorarium gene sequences. By computational treatment, the observed mutation events in the gene frameshifting sub-sequence have been neutralised. Each nucleotide insertion must be eliminated and each nucleotide deletion must be substituted by the symbol N (any nucleotide). When the frameshifting fragments of the amino acid sequences were substituted by the computationally neutralised subsequences, the beta-glucosidase alignments were improved. We also discuss the structural implications of the compensated frameshift mutations events. PMID- 14527733 TI - Association of HLA class I alleles with psoriasis vulgaris in southeastern Chinese Hans. AB - BACKGROUND: Predisposing genetic factors in psoriasis include associations with human leukocyte antigen (HLA). Accumulative evidence has shown that certain HLA at class I locus, especially HLA-Cw6, are associated closely with psoriasis. OBJECTIVE: To evaluate the association of HLA class I alleles with susceptibility to psoriasis in the southeastern Chinese Han population. METHODS: We performed genotype for HLA-A, -B and -C loci in 166 patients with psoriasis vulgaris by means of polymerase chain reaction with sequence-specific primers technique. The distribution of HLA allelic frequencies was further analyzed according to age of onset, i.e. under 35-y and beyond 35-y groups. These data were compared with the healthy controls of 204 unrelated Hans. RESULTS: The frequencies of HLA-A*26 (24.7% vs. 13.1%, OR=2.36, Pc<0.01), -B*13 (27.2% vs. 14.8%, OR=2.34, Pc<0.01), B*27 (12.2% vs. 4.0%, OR=3.49, Pc<0.01) and -Cw*0602 (17.9% vs. 5.3%, OR=4.20, Pc<0.001) were significantly increased in psoriasis patients, whereas HLA-Cw*0304 frequency (4.9% vs. 13.4%, OR=0.32, Pc<0.01) was highly decreased, when compared to the controls. HLA-A*26-B*27-Cw*0602 was identified as a high-risk haplotype of HLA class I in developing psoriasis in the test. HLA-Cw*0602 was found to be strongly associated with the early-onset psoriasis (age of onset <35 y). CONCLUSION: This study demonstrated the positive associations of HLA class I markers with psoriasis vulgaris, of which HLA-Cw*0602 was the strongest susceptibility determinant for development of early-onset psoriasis, in the southeastern Chinese Han population. PMID- 14527734 TI - Downregulation of TGFbeta isoforms and their receptors contributes to keratinocyte hyperproliferation in psoriasis vulgaris. AB - BACKGROUND: Psoriasis vulgaris is a chronic inflammatory disorder characterized by epidermal hyperproliferation. Transforming growth factor beta (TGFbetas) have a major antiproliferative action in epidermis. OBJECTIVE: We evaluated the distribution and levels of expression of TGFbeta isoforms and their receptors in psoriatic versus normal skin with the goal of discovering potential alterations in TGFbeta signal transduction associated with psoriasis. METHODS: Expression of TGFbeta isoforms and their receptors was analyzed in normal and psoriatic skin using immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR) techniques. Furthermore, DNA synthesis was measured in normal keratinocytes transfected with a dominant-negative TGFbeta receptor II (TbetaRII) vector that eliminated most of the cytoplasmic TbetaRII domain. RESULTS: Marked elevations in DNA synthesis, as assessed by BrdU incorporation and proliferating cell nuclear antigen (PCNA) immunoreactivity, were confirmed in psoriatic epithelial cells. Using immunohistochemistry and RT-PCR analysis, expression of TGFbeta2 and 3 was diminished in the psoriatic epidermis as compared with those observed in normal skin. With respect to TGFbeta receptors, expression of TbetaRI and II was markedly decreased in the psoriatic epidermis. In addition, levels of Smad2 mRNA were also decreased in psoriatic skin. Transfection of normal keratinocytes with the dominant-negative TbetaRII vector significantly elevated DNA synthesis as compared with keratincoytes transfected with control vector (under condition of TGFbeta addition), suggesting that the dominant-negative TbetaRII mutant inhibits the antiproliferative effects of TGFbeta. CONCLUSION: The present investigation strongly suggest that the TGFbeta signaling pathway is downregulated in psoriatic skin and this situation leads to abnormal cell proliferation due to a functional decrease in growth regulation. PMID- 14527735 TI - Incidence of cytokeratin 20 expressed cells in primitive follicular structure and secondary neoplastic proliferations of nevus sebaceus. AB - BACKGROUND: Nevus sebaceus of Jadassohn (NSJ) is a complex hamartoma in which various kinds of secondary neoplastic or hyperplastic proliferations such as primitive follicular structure (PFS) may arise. Recently, cytokeratin (CK) 20 expressed cells in PFSs and trichoblastomas of NSJs were demonstrated. OBJECTIVE: To clarify the nature and relationship among these proliferations, those in other secondary neoplasms and precise CK distributions in such lesional epidermis are examined. METHODS: 49 cases of NSJ were analyzed clinicopathologically and immunohistochemically using ten monoclonal antibodies against involucrin and CKs. RESULTS: 15 of 49 cases possessed PFSs, and six cases, of which five were associated with PFSs, possessed seven lesions of secondary neoplasms: four BCC, two syringocystadenoma papilliferum (SAP) and one trichilemmoma. The age distribution of 15 patients with PFSs was significantly higher than that of the other 34 patients without the PFSs. The six patients with secondary neoplasms were statistically situated in a higher age range than the ten cases with the PFSs alone. Although the CK expression in lesional epidermis of NSJs were similar to that of normal epidermis, CK20 expressed cells were scattered in the PFSs of 11 cases among the 15 NSJs, but not in the other four cases. Three of the four cases were associated with BCC in which CK20 was not expressed. CONCLUSIONS: From the statistical analysis, we presumed that the existence of the PFSs indicates potency to develop secondary neoplasms. Moreover, the absence of CK20 expressed cells (Merkel cells) in the structures was speculated to predict the development of BCC in the future. PMID- 14527736 TI - Effective treatment of pruritus in atopic dermatitis using H1 antihistamines (second-generation antihistamines): changes in blood histamine and tryptase levels. AB - BACKGROUND: Atopic dermatitis (AD) is a common chronic inflammatory and allergic skin disease that almost always begins in childhood and follows a course of remittance and flare-up. AD is characterized by intense pruritus and itchiness that can be triggered by an interplay of genetic, immunologic and environmental factors. Of the mediators, histamine is one of the most potent inducers of pruritus. Serum tryptase, which is also a mediator, may be used to examine allergic disease as well. The development of minimal sedation H1 antihistamines (second-generation antihistamines) has revolutionized treatment of allergic diseases. OBJECTIVE: The present study examines the efficacy of second-generation antihistamines in relieving pruritus due to AD. In addition, the relationship between AD pruritus and antihistamine therapy was analyzed by measuring the blood histamine and tryptase levels. METHODS: Thirty-two AD patients were recruited and underwent second-generation antihistamine therapy for 2 weeks. Seventeen received combined topical corticosteroid treatment (Group 1) and the other 15 did not receive steroid treatment (Group 2). The Severity Index and Pruritus Score were assessed as an AD clinical activity index and compared with baseline data. RESULTS: Both the Severity Index and Pruritus Score improved significantly in Group 1 (P<0.001, P<0.05). Group 2 demonstrated a significant improvement in Pruritus Score (P<0.05), but not in the Severity Index. Plasma histamine levels were significantly higher in AD at baseline compared with healthy controls. CONCLUSION: Following antihistamine therapy, these levels decreased significantly in both AD groups (P<0.05). There was a significant correlation between baseline blood histamine and typtase levels. However, this correlation was not evident following treatment. This may reflect insufficient detection capabilities of the measuring assay. The present results suggest that second-generation antihistamine therapy provides an effective clinical treatment for AD, with a notable improvement in pruritus. Furthermore, antihistamine therapy reduced plasma histamine levels in AD patients. These findings further suggest that high blood histamine and tryptase levels in AD patients contribute to the pathogenesis of this disorder, including the onset of pruritus. PMID- 14527737 TI - Up-regulation of interleukin-13 receptor alpha1 on human keratinocytes in the skin of psoriasis and atopic dermatitis. AB - BACKGROUND: Interleukin (IL)-13 is a pleiotropic cytokine, which shares many biological functions with IL-4. The receptor subunits of IL-13 consist of IL 4Ralpha, IL-13Ralpha1 and IL-13Ralpha2. The regulatory mechanisms of the IL 13Ralpha expression in the keratinocytes of certain skin disease have not been known. OBJECTIVE: To clear the roles of IL-13 and the regulatory mechanisms of its receptor in atopic dermatitis (AD) and psoriasis. METHOD: The expression of IL-13Ralpha1 in the skin of AD and psoriasis was investigated by immunohistochemistry. The regulation of IL-13Ralpha mRNA in the skin and human primary keratinocyte (HPK) was investigated by quantitative PCR. The secretion of IL-6 and RANTES from HPK was measured by ELISA. RESULTS: The expression of IL 13Ralpha1 was more prominent on the suprabasal keratinocytes in the skin of AD and striking increase of staining was observed on all layers of keratinocyte in the skin of psoriasis. The mRNA of IL-13Ralpha1, but not of IL-13Ralpha2 was overexpressed in both skin of AD and psoriasis. In vitro experiment using HPK demonstrated that IFN-gamma, IL-13 but not IL-4 could up-regulate the mRNA expression of IL-13Ralpha1. In contrast, IL-13Ralpha2 mRNA expression was up regulated by IFN-gamma plus IL-4. Furthermore, the stimulation of HPK with IFN gamma plus IL-13 and/or IL-4 resulted in significant enhancement of IL-6 and RANTES secretion. CONCLUSION: These findings indicate that IL-4 and IL-13 have different regulatory effects on the expression of IL-13Ralpha1 and alpha2, and the overexpression of IL-13Ralpha1 may play some roles in the pathogenesis of chronic stage of AD or psoriasis. PMID- 14527738 TI - Species-identification of dermatophytes Trichophyton, Microsporum and Epidermophyton by PCR and PCR-RFLP targeting of the DNA topoisomerase II genes. AB - BACKGROUND: We have focused on the DNA topoisomerase II genes of pathogenic fungi and have previously applied polymerase chain reaction (PCR)-based identification of several species including the some of the major dermatophyte species. OBJECTIVE: To identify the dermatophytes (18 species) to a species level by PCR and PCR-restriction fragment length polymorphism (RFLP) techniques, without determining the nucleotide sequence. METHODS: The genomic DNAs of the dermatophytes (ten species of Trichophyton, seven species of Microsporum, and Epidermaphyton floccosum) were amplified by PCR using a common primer set (dPsD1) for the dermatophytes, followed by nested PCR using other primer sets (dPsD2, PsT and PsME) that contained primers specific for the DNA topoisomerase II genes of the dermatophytes. PCRs using PsT and PsME were used for the species identification of Trichophyton, Microsporum and E. floccosum. The PCR products generated by dPsD2 were digested with restriction enzymes (Hinc II, Hinf, Afl II and PflM I), and the restriction profiles were analyzed. RESULTS: Of the eighteen species of dermatophytes, five species (T. rubrum, T. violaceum, M. canis, M. gypseum and E. floccosum) were specifically identified by the PCR using PsT and PsME to the species level, and the remaining species were identified by the unique restriction profiles for each species in the PCR-RFLP analysis, except that the restriction profile of T. mentagrophytes var. interdigitale was identical to that of T. mentagrophytes var. quinckeanum. CONCLUSION: PCR and PCR RFLP techniques targeting the DNA topoisomerase II gene are simple and rapid, and quite useful as tools for the identification of dermatophytes to the species level. PMID- 14527739 TI - Neuropeptides concentrations in the skin of a murine (NC/Nga mice) model of atopic dermatitis. AB - BACKGROUND: It has been reported that the expression of neuropeptides (NPs), and the density and structure of peripheral nerves in atopic dermatitis (AD) are different from those in normal skin. OBJECTIVE: We investigated the role of NPs, in the development of AD with quantitative study of substance P (SP) and calcitonin gene-related peptide (CGRP) in the skin of AD-model mice. METHODS: We measured the NPs in the skin of mice (NC/Nga as AD-model mice, BALB/c and C57BL/6 as control) by enzyme-linked immunosorbentassay (ELISA). Peripheral nerve fibers and SP in the skin were stained by immunohistochemical staining, using anti PGP9.5 antibody and anti-SP antibody. RESULTS: Under conventional condition, SP concentration in AD-like skin lesions of NC/Nga mice was higher than that in non affected skin of the same mice. Under specific pathogen-free condition, SP concentration in the skin of NC/Nga mice was higher than that in the skin of BALB/c and C57BL/6 mice. In contrast, CGRP concentration in the skin lesions was lower than that in non-affected skin of NC/Nga mice. SP was detected not only in the nerve fibers in the dermis but also in mast cells in the inflammatory areas. CONCLUSIONS: The skin of NC/Nga mice contains more SP congenitally, and environmental factors may aggravate this abnormal condition. We hypothesize that increase of SP accompanied with a decrease of CGRP in the skin may play important roles in the pathogenesis and development of AD. PMID- 14527740 TI - A unique point mutation in the NSDHL gene in a Japanese patient with CHILD syndrome. PMID- 14527741 TI - Serum platelet-activating factor acetylhydrolase activity in patients with atopic dermatitis. PMID- 14527742 TI - Detection of Sporothrix schenckii chitin synthase 1 (CHS1) gene in biopsy specimens from human patients with sporotrichosis. PMID- 14527743 TI - No associations of CTLA-4 and ICAM-1 polymorphisms with psoriasis in the Korean population. PMID- 14527744 TI - Mathematical modeling of human granulopoiesis: the possible importance of regulated apoptosis. AB - Steady state human granulopoiesis was modeled by a convection-reaction differential equation of the Rubinow type for the bone marrow granulocyte precursors and an ordinary differential equation for the blood granulocytes. Measured values reported from several laboratories were used as sources for the model proliferation, maturation, and mobilization rates. Due to the large variability in the measured input data, four alternative models were constructed initially, each one with a specific combination of proliferation rate and maturation rate. They were all able to produce output values for the bone marrow neutrophil count and turnover rate close to accepted data, but neither of them could reproduce good values for the differential fractions of the neutrophil precursor stages. The model output was especially sensitive to changes in transit time in the mitotic relative to the postmitotic precursor compartments. When the net proliferation rate was modeled to optimize the bone marrow differential fractions according to published data, the total bone marrow neutrophil count would not fit with published data. However, a composite model optimizing differential fractions, bone marrow neutrophil count, and turnover rate yielded plausible output values and a reduced proliferation rate in the myelocyte stage. This result opens for a possibly substantial apoptosis rate at the myelocyte stage in accordance with results from earlier investigators. However, the result was based on a special choice of precursor transit times, taken from the literature. More precise data concerning granulocyte precursor cycle times, transit times, and differential fractions would radically improve the model's ability to clarify the role of apoptosis during granulocyte production and storage. PMID- 14527745 TI - A mechanistic mathematical model of temozolomide myelosuppression in children with high-grade gliomas. AB - Temozolomide (TMZ) is currently being evaluated for the treatment of high-grade gliomas in children. Myelosuppression (the suppression of bone marrow activity) is the dose-limiting toxicity for TMZ in adults and children. Empirical methods (i.e. relations between the percent change in absolute neutrophil count (ANC) and the area under the plasma concentration curve (AUC) of TMZ or its active metabolite MTIC) showed poor results when attempting to describe myelosuppression from serial data derived during TMZ therapy in a Phase II study of children with high-grade glioma. Therefore, to improve our understanding of the myelosuppressive effects of TMZ and MTIC in children we developed a mechanistic mathematical model. The model describes the progression of neutrophils from their production in the bone marrow to their release in the plasma. Included in the model are the feedback effects of granulocyte colony stimulating factor (G-CSF), which stimulates neutrophil production when there is a decrease in circulating neutrophils. The model is fit to serial ANC measurements obtained after TMZ dosing and it is able to explain, among other things, the lag in ANC reduction following a dose of TMZ, the ANC nadir, and the 'rebound effect' observed where the ANC recovers to levels greater than that observed pre-TMZ dose. This model will be useful for the prospective design of clinical trials of TMZ in children with cancer. PMID- 14527746 TI - A cylindrical model for studying subendocardial ischaemia in the left ventricle. AB - In this paper a mathematical model of a left ventricle with a cylindrical geometry is presented with the aim of gaining a better understanding of the relationship between subendocardial ischaemia and ST depression. The model is formulated as an infinite cylinder and takes into account the full bidomain nature of cardiac tissue, as well as fibre rotation. A detailed solution method (based on Fourier series, Fourier transforms and a one dimensional finite difference scheme) for the governing equations for electric potential in the tissue and the blood is also presented. The model presented is used to study the effect increasing subendocardial ischaemia has on the epicardial potential distribution as well as the effects of changing the bidomain conductivity values. The epicardial potential distributions obtained with this cylindrical geometry are compared with results obtained using a previously published slab model. Results of the simulations presented show that the morphologies of the epicardial potential distributions are similar between the two geometries, with the main difference being that the cylindrical model predicts slightly higher potentials. PMID- 14527747 TI - Non-linear, non-invasive method for seizure anticipation in focal epilepsy. AB - In this paper we discuss an approach, using methods of non-linear time series analysis applied to scalp electrode recordings, which is able to distinguish between epochs temporally distant from and just prior to, the onset of a seizure in patients with temporal lobe epilepsy. The method involves a comparison of recordings taken from electrodes adjacent to and remote from the site of ictal onset. In particular, we define a non-linear quantity which we call 'marginal predictability'. This quantity is computed using data from remote and from adjacent electrodes. We find that the difference between the marginal predictabilities computed for the remote and adjacent electrodes decreases several tens of minutes prior to seizure onset, compared to its value interictally. PMID- 14527748 TI - An exactly solvable model of population dynamics with density-dependent migrations and the Allee effect. AB - We consider a single-species model of population dynamics allowing for migrations and the Allee effect. Two types of migration are taken into account: one caused by environmental factors (e.g., a passive transport with the wind or water current) and the other associated with biological mechanisms. While the first type is apparently density-independent, the speed of migration in the second one can depend on the population density. Mathematically, this model consists of a non-linear partial differential equation of advection-diffusion-reaction type. Using an appropriate change of variables, we obtain an exact solution of the equation describing propagation of travelling population fronts. We show that, depending on parameter values and thus on the relative intensity of density dependent and density-independent factors, the direction of the propagation can be different thus describing either species invasion or species retreat. PMID- 14527749 TI - Mathematical analysis of a model describing evolution of an asexual population in a changing environment. AB - We investigate a mathematical model for an asexual population with non overlapping (discrete) generations, that exists in a changing environment. Sexual populations are also briefly discussed at the end of the paper. It is assumed that selection occurs on the value of a single polygenic trait, which is controlled by a finite number of loci with discrete-effect alleles. The environmental change results in a moving fitness optimum, causing the trait to be subject to a combination of stabilising and directional selection. This model is different from that investigated by Waxman and Peck [Genetics 153 (1999) 1041] where overlapping generations and continuous effect alleles were considered. In this paper, we consider non-overlapping generations and discrete effect alleles. However in [Genetics 153 (1999) 1041] and the present work, there is the same pattern of environmental change, namely a constant rate of change of the optimum. From [Genetics 153 (1999) 1041], no rigorous theoretical conclusion can be drawn about the form of the solutions as t grows large. Numerical work carried out in [Genetics 153 (1999) 1041] suggests that the solution is a lagged travelling wave solution, but no mathematical proof exists for the continuous model. Only partial results, regarding existence of travelling wave solutions and perturbed solutions, have been established (see [Nonlin. Anal. 53 (2003) 683; An integral equation describing an asexual population in a changing environment, Preprint]). For the discrete case of this paper, under the assumption that the ratio between the unit of genotypic value and the speed of environment change is a rational number, we are able to give rigorous proof of the following conclusion: the population follows the environmental change with a small lag behind, moreover, the lag is represented using a calculable quantity. PMID- 14527750 TI - The use of the palliative prognostic score in patients with diagnoses other than cancer. PMID- 14527751 TI - Re: Use of IM narcotic injections in hospitalized patients. PMID- 14527753 TI - Can we really consider quality of life as an outcome of palliative care? PMID- 14527756 TI - Re: High dose opioids in pediatric palliative care. PMID- 14527757 TI - Quality of life and psychosocial issues in ventilated patients with amyotrophic lateral sclerosis and their caregivers. AB - Non-invasive ventilation (NIV) is an efficient palliative measure for symptoms of chronic hypoventilation in patients with amyotrophic lateral sclerosis (ALS), and can also lengthen survival. A subset of ALS patients undergoes tracheostomy ventilation (TV) for life prolongation. We investigated the quality of life (QOL) and psychosocial situation of 52 home ventilated ALS patients and their caregivers. The battery included sociodemographic, generic, and disease-specific variables, as well as the Profile of Mood States and the Munich Quality of Life Dimensions List. Data were compared between the NIV (n=32) and the TV (n=21) groups. Mean ventilation time was 14 months for NIV and 35 months for TV. Eighty one percent of TV patients had been tracheotomized without informed consent. The data show a good overall QOL for both NIV and TV patients, but a very high burden of care for TV caregivers, 30% of whom rated their own QOL lower than their patient's QOL. Sexuality was an important issue. Thus, any assessment of QOL in a home palliative care situation should include the primary caregivers. PMID- 14527758 TI - Respiratory tract secretions in the dying patient: a retrospective study. AB - Respiratory tract secretions (RTS), the sound created by poorly-cleared mucous in the hypopharynx or bronchial tree, can be alarming for dying patients, relatives and staff. Increased knowledge into the etiology of RTS and its response to treatment is needed to improve future management. We studied retrospectively the data from 202 patients who died on a 30-bed specialist palliative care unit during a one-year period. These patients were observed every four hours during the dying phase. RTS was treated with hyoscine hydrobromide. Ninety-nine patients (49%) developed RTS. The median time from onset of RTS till death was 16 hours. Fifty-nine patients could have their treatment response assessed. Of these, 30.5% responded within four hours, 33.9% after four hours, and 35.5% died with RTS. Increasing the dose for nonresponders had no significant effect. Significant risk factors for developing RTS were found to be prolonged dying phase, primary lung cancer and male gender. PMID- 14527759 TI - Pain and its relation to cognitive function and depressive symptoms: a Swedish population study of 70-year-old men and women. AB - The aim of this study was to investigate the prevalence of pain and its characteristics, and to examine the association of pain with cognitive function and depressive symptoms, in a representative sample of 70-year-old men and women. Data were collected within the gerontological and geriatric population studies in Goteborg, Sweden (H-70). A sample of 124 men and 117 women living in the community took part in the study. A questionnaire was applied which included four different aspects of pain experience: prevalence, frequency of episodes of pain, duration and number of locations. In close connection to this, depressive symptoms were assessed using the Center for Epidemiological Studies Depression Scale. The prevalence of pain during the last 14 days was higher in women (79%; n=91) than in men (53%; n=65) (P<0.001). Women (68%; n=78) also reported pain that had lasted for >6 months to a greater extent than men (38%; n=46) (P<0.001). The frequency of episodes of pain was also higher among women, 64% (n=74) reporting daily pain or pain several days during the last 14 days while 37% of the men (n=45) did so (P<0.001). Women (33%, n=38) also reported pain experience from >/=3 locations more often than men (11%; n=13) (P<0.001). On the other hand, the association between depressive symptoms and pain experience was more evident in men than in women. Women were taking significantly more antidepressants compared to men (P<0.03). The results show that pain is common in 70-year-old people and especially in women. However, associations between depressive symptoms and the four aspects of pain experience were more pronounced among men. PMID- 14527760 TI - Neuropsychological effects of long-term opioid use in chronic pain patients. AB - Opioids are thought to worsen the performance of psychomotor tasks due to their sedating and mental-clouding effects. As a result, some safety regulations currently restrict the use of opioids when driving or using heavy equipment. We investigated the psychomotor effects of long-term opioid use in 144 patients with low back pain. All subjects were administered two neuropsychological tests (Digit Symbol and Trail Making Test-B) before being prescribed opioids for pain; tests were re-administered at 90- and 180-day intervals. Test scores significantly improved while subjects were taking opioids for pain, which suggests that long term use of oxycodone with acetaminophen or transdermal fentanyl does not significantly impair cognitive ability or psychomotor function. PMID- 14527761 TI - Care for the caregivers: a review of self-report instruments developed to measure the burden, needs, and quality of life of informal caregivers. AB - Significant demands are being placed on the informal caregivers of chronically ill patients, including those suffering from cancer. Health care professionals need to be aware of these demands, and they need effective tools to assess the impact these demands place on the caregivers. Over the past 25 years, researchers have developed self-report instruments to assess informal caregivers. These instruments assess various aspects of the caregiving experience, including caregiver burden, needs, and quality of life. The purpose of this review was to identify and critically evaluate these instruments. MEDLINE and PUBMED were searched from 1966 to 2002. After an extensive literature search and review, and utilizing specific inclusion criteria, 28 instruments were identified and evaluated in terms of their development, content, and psychometric properties. In addition, a history of the construct and measurement development in the areas of caregiver burden, needs, and quality of life are discussed. Although some further development and refinement of instruments could benefit the field, depending on the questions researchers or clinicians seek to pursue, there are many proven tools available for their use. Future research needs to use these instruments to assess the effectiveness of interventions aimed at improving the care of the caregivers. PMID- 14527767 TI - Proceedings of the International Symposium Hot Topics in Urinary Tract Infections, part 1. 25 January 2003, Budapest, Hungary. PMID- 14527768 TI - Virulence factors of uropathogenic Escherichia coli. AB - Virulence factors of Escherichia coli are of two main types; those produced on the surface of the cell and those produced within the cell and then exported to the site of action. Those on the surface include different sorts of fimbriae that have a role in adhesion to the surface of host cells but may also have additional roles such as tissue invasion, biofilm formation or cytokine induction. The activities of cell wall components are discussed and several exported virulence factors are described that have anti host cell activities. Others virulence factors enable the bacteria to grow in an environment of iron restriction. PMID- 14527769 TI - Pathogenesis and management of bacterial urinary tract infections in adult patients with diabetes mellitus. AB - Urinary tract infections (UTIs) are more common and tend to have a more complicated course in patients with diabetes mellitus (DM). The mechanisms, which potentially contribute to the increased prevalence of both asymptomatic and symptomatic bacteriuriai in these patients are defects in the local urinary cytokine secretions and an increased adherence of the microorganisms to the uroepithelial cells. The need for treatment of asymptomatic bacteriuria remains controversial. No evidence is available on the optimal treatment of acute cystitis and pyelonephritis in patients with DM. Because of the frequent (asymptomatic) upper tract involvement and the possible serious complications, many experts recommend a 7-14-day oral antimicrobial regimen for bacterial cystitis in these patients, with an antimicrobial agent that achieves high levels both in the urine and in urinary tract tissues. Current data suggest that shorter regimens will lead to failure also in uncomplicated UTI in women. The recommended treatment of acute pyelonephritis does not differ from that in nondiabetic patients. Clinical trials specifically dealing with the treatment of UTIs in diabetic patients, comparing the optimal duration and choice of antimicrobial agent, are needed. Besides that, new approaches to preventive strategies must prove their value in this specific patient group. PMID- 14527770 TI - Asymptomatic bacteriuria. Clinical significance and management. AB - The clinical significance and management of asymptomatic bacteriuria (ASB) differs according to different groups of patients. ASB requires antibiotic treatment in pregnant women, children aged 5-6 years and prior to invasive genitourinary procedures. However, there is a consensus that ASB in the elderly, healthy school girls and young women, diabetic women and patients with indwelling catheters or intermittent catheterization has no clinical significance and antibiotic prescription is not indicated. PMID- 14527771 TI - Prevalence and antimicrobial susceptibility of pathogens in uncomplicated cystitis in Europe. The ECO.SENS study. AB - The ECO.SENS study investigated the prevalence and antimicrobial susceptibility of pathogens causing community-acquired acute uncomplicated urinary tract infections (UTIs) in 4734 women aged 18-65 years presenting with symptoms of acute UTI, at 252 community healthcare centres in 16 countries in Europe plus Canada. Resistance in Escherichia coli occurred most frequently to ampicillin (30%) and sulphonamides (29%), followed by trimethoprim (15%), trimethoprim/sulphamethoxazole (14%) and nalidixic acid (5%) but was low to co amoxiclav, mecillinam, cefadroxil, nitrofurantoin, fosfomycin, gentamicin and ciprofloxacin, all at<3%. Consumption of antibiotics in 1997 varied more than 4 fold within Europe and from 9 to 37 DDD/1000 inhabitants/day, the consumption being highest in Southern Europe. The consumption of broad-spectrum penicillins correlated with resistance to ampicillin and there was a clear correlation between quinolone consumption and resistance to ciprofloxacin and nalidixic acid. The 4-fold difference in antibiotic consumption within Europe and the correlation to resistance emphasises the importance of controlling antibiotic usage. PMID- 14527772 TI - In vitro activity of fosfomycin against gram-negative urinary pathogens and the biological cost of fosfomycin resistance. AB - The aim of this study was to reassess the activity of fosfomycin against recently isolated uropathogens circulating in Italy and to evaluate the effect of fosfomycin resistance on the expression of several virulence traits using the rare mutant strains. In vitro activity of fosfomycin was evaluated using 441 Gram negative organisms isolated from patients with uncomplicated urinary tract infections (UTIs). Fosfomycin was the most active antibiotic against Escherichia coli (99% susceptibility). The activity against Proteus mirabilis was more potent than that of co-trimoxazole and nitrofurantoin (87.5, 67 and 0% susceptibility, respectively). The other microorganisms, accounting for about 7% of all pathogens tested, showed variable susceptibilities to fosfomycin. Compared with susceptible strains, fosfomycin-resistant mutants showed a reduced rate of growth and were impaired in their ability to adhere to uroepithelial cells and to urinary catheters. They were also more resistant to UV irradiation and to phage T7 and showed diminished rates of colicin synthesis and transfer of plasmids. PMID- 14527773 TI - Urinary tract infections in South Croatia: aetiology and antimicrobial resistance. AB - The aim of this study was to determine the distribution of uropathogens isolated from outpatients living in South Croatia and the in vitro susceptibility of these organisms to antimicrobial agents. Of 5080 enrolled uropathogen isolates, 82.28% isolates were Gram-negative, the most frequent isolates being Escherichia coli (62.62%), enterococci (10.18%), Proteus mirabilis (5.31%), Streptococcus agalactiae (3.84%), Staphylococcus spp. (3.70%), Pseudomonas spp. (3.46%), Klebsiella spp. (2.38%). The E. coli resistance rate was 42.17% to amoxycillin, 20.59% to trimethoprim-sulphamethoxazole and 6.09% to norfloxacin. Almost all Klebsiella spp. isolates were resistant to amoxycillin and the resistance rate to trimethoprim-sulphamethoxazole was over 20%, and 14.15% to the fluoroquinolones. A high methicillin-resistance rate was found among S. aureus (61.22%) and coagulase negative staphylococci (41.48). PMID- 14527774 TI - Fluoroquinolones and resistance in the treatment of uncomplicated urinary tract infection. AB - Acute uncomplicated cystitis is one of the most common problems for which young women seek medical attention. Most of these infections are caused by Escherichia coli which are susceptible to many oral antimicrobials, although resistance is increasing to some of the commonly used agents, especially trimethoprim/sulphamethoxazole (TMP/SMX). In women with risk factors for infection with resistant bacteria, or in the setting of a high prevalence of TMP/SMX resistance, a fluoroquinolone or nitrofurantoin should be considered for empirical treatment. Use of nitrofurantoin does not share cross-resistance with more commonly prescribed antimicrobials and its more widespread use is justified from a public health perspective as a fluoroquinolone-sparing agent. beta-lactams and fosfomycin should be considered second-line agents for empirical treatment of cystitis. For acute uncomplicated pyelonephritis, fluoroquinolones are superior to TMP/SMX for empirical therapy due to the relatively high prevalence of TMP/SMX resistance among uropathogens causing pyelonephritis. TMP/SMX, effective for patients with mild to moderate disease, is an appropriate drug if the uropathogen is known to be susceptible. It is reasonable to use a 7-10 day oral fluoroquinolone regimen for outpatient management of mild to moderate pyelonephritis in the setting of a susceptible causative pathogen and rapid clinical response to therapy. Most women with acute uncomplicated pyelonephritis are now managed safely and effectively as outpatients. PMID- 14527775 TI - Pivmecillinam--therapy of choice for lower urinary tract infection. AB - Pivmecillinam is the pro-drug of mecillinam, a beta-lactam antibiotic with a novel site of action and with specific and high activity against Gram-negative organisms such as Escherichia coli and other Enterobacteriaceae. Since its introduction, it has been widely used for the treatment of acute lower urinary tract infections (UTI), primarily in the Nordic countries. In contrast to the increasing resistance of urinary pathogens to other beta-lactams particularly ampicillin/amoxycillin and to other UTI antibiotics such as trimethoprim and trimethoprim/sulphamethoxazole (TMP/SMX), the level of resistance has remained on a low level. Less than 2% of E. coli community isolates are resistant to mecillinam. This paper reviews the clinical data on pivmecillinam with a special focus on the safety aspects. A large number of studies from the 70s to 80s have proven the clinical efficacy and safety of pivmecillinam for empirical treatment of acute cystitis. More recent studies confirm short-term treatment with pivmecillinam results in clinical and bacteriological cure rates similar to those obtained with other UTI agents. Both clinical studies in pregnant women with UTI and large epidemiological studies have confirmed the safety of pivmecillinam used in pregnancy. In the Nordic countries pivmecillinam has been the most widely used agent for treatment of UTI in pregnancy for many years. Ecological aspects of antibiotic treatment are important both with regard to adverse effects and development of resistance due to disturbance of the normal micro flora. Studies have shown that pivmecillinam has a very minor impact on the normal oropharyngeal, intestinal and skin microflora. The clinical implications of this are a low frequency of diarrhoea and Candida vaginitis as confirmed in the clinical studies. The high and increasing level of resistance among E. coli to currently recommended first-line agents for acute cystitis requires a re evaluation of treatment guidelines. With the low resistance, its proven efficacy and favourable safety profile, pivmecillinam is a suitable first-line agent for empirical treatment of acute cystitis. PMID- 14527776 TI - Why fosfomycin trometamol as first line therapy for uncomplicated UTI? AB - Uncomplicated urinary tract infection (UTI) is one of the most common conditions requiring diagnostic and therapeutic intervention. The aetiology and the treatment of these infectious diseases have changed little during last years of the 'antibiotic era'. Escherichia coli is the most prevalent uropathogen (85 >90%) and treatment is aimed at eradicating the infection using shorter regimes that typically may employ a 3-day course with once-a-day dosing of a selected drug or a single dose of a particular efficacious antibiotic. Antibiotic resistance to commonly used agents, such as trimethoprim and ampicillin, often now exceeds 30-50%, while fosfomycin trometamol, despite many years of usage, continues to be characterized by an extremely low incidence of E. coli resistant strains (about 1%) worldwide. Many factors may have contributed to preserve fosfomycin trometamol antibacterial activity including single dose usage limited to urinary infections, very high and sustained urinary concentrations that rapidly kill bacteria reducing the opportunity for mutant selection. In addition there is no animal feed that contains the drug, resistance is most commonly acquired by chromosomal mutations that do not spread easily and the biological cost of these genetic modifications is high. To these parameters fosfomycin trometamol adds excellent tolerability and safety. Although nowadays, microbial resistance limits available resources and some drugs can no longer be recommended as reliable agents, fosfomycin trometamol, because of its properties, remains a drug of choice for the eradication of uncomplicated UTI. PMID- 14527777 TI - Short term therapy for uncomplicated urinary tract infection today. Clinical outcome upholds the theories. AB - The clinical and bacteriological activity of fosfomycin trometamol (FT) has been compared with several other antibiotics in the treatment of uncomplicated urinary tract infections. A single dose of FT had activity comparable with a 5-day course of trimethoprim in a trial where the causative organism and its sensitivity were unknown. In another trial FT showed better long term eradication compared with a 5-day course of cephalexin and other studies suggested a single dose of FT was comparable with a 7-day course of nitrofurantoin or norfloxacin. There were few important side effects with fosfomycin therapy and it was considered a safe and effective first line treatment in uncomplicated urinary tract infection. PMID- 14527778 TI - Febrile urinary tract infection in men. AB - Febrile urinary tract infection (UTI) in adult men has received little attention in the literature, partly due to the low incidence compared with women. In a multidisciplinary project, infectious disease specialists, urologists and microbiologists studied the host-parasite interaction in men from Gothenburg with community-acquired febrile UTI. The results are discussed in the light of other similar studies and related to findings in women. Contrary to the traditional view, men and women with febrile UTI have very similar infecting bacterial species, host predispositions and treatment results. However, virulence expression differs in the infecting Escherichia coli in both sexes, besides the overt anatomical distinction including having a prostate. PMID- 14527779 TI - Effect of fosfomycin alone and in combination with N-acetylcysteine on E. coli biofilms. AB - Four slime-producing uropathogenic Escherichia coli strains were used to investigate the activity of fosfomycin and N-acetylcysteine (NAC) against biofilms developed on 96-well polystyrene tissue culture plates. Biofilms aged, respectively, 5 (initial) and 48 h (mature) and two fosfomycin concentrations (128 and 2000 mg/l) were used. The effect of various levels (0.007-8 mg/ml) of NAC alone and in combination with fosfomycin on the formation or disruption of biofilms was assessed. Following exposure to the drugs, the percentage of residual slime relative to the control, ranged from 62.5-100 to 26.2-64.1% in the presence of 0.007 and 8 mg/ml of NAC. After treatment of pre-formed biofilms with NAC at the highest concentrations used, the remaining exopolysaccharide matrix was reduced to 25-68% of the amount found with the untreated control. Exposure to fosfomycin at 2000 mg/l reduced biofilms 40-57 and 41-49% for the initial and mature forms, respectively. Fosfomycin was more active at 2000 mg/l combined with NAC 2 mg/ml. Under these conditions initial and mature biofilms were reduced 66 80 and 60-73%, respectively. NAC, when used in combination, enhanced fosfomycin bactericidal activity producing a 99-99.9% reduction in viable cells. Fosfomycin and NAC at concentrations achievable in urine displayed a synergistic effect promoting both the formation of biofilms and reduction of sessile cell viability. PMID- 14527780 TI - High sucrose concentration protects E. coli against high pressure inactivation but not against high pressure sensitization to the lactoperoxidase system. AB - The inactivation of Escherichia coli by high hydrostatic pressure treatment at up to 550 MPa and 20 degrees C was studied in potassium phosphate buffer containing high concentrations of sucrose. E. coli strain MG1655 was pressure-sensitive in the absence of sucrose, but became highly pressure resistant in the presence of 10% to 50% (w/v) sucrose. The pressure resistance of E. coli strain LMM1010, a previously described derivative of MG1655 that is pressure resistant in the absence of sucrose, was further increased in the presence of sucrose, to a similar level as for strain MG1655 in the presence of sucrose. When cell suspensions of either strain were stored after pressure treatment for 24 h at 20 degrees C, a further reduction of the plate counts indicative of pressure induced sublethal injury was observed, that was positively correlated with pressure intensity and negatively with sucrose concentration. Addition of the lactoperoxidase system to the cell suspensions strongly enhanced high pressure inactivation of E. coli at high sucrose concentrations. Using a pressure intensity of only 250 MPa, both E. coli strains were sensitized for the lactoperoxidase system in up to 30% (w/v) sucrose, resulting in at least 10(6) fold inactivation within 24 h or less after pressure treatment. For comparison, a pressure treatment at 250 MPa in the absence of the lactoperoxidase system did not cause any inactivation of either strain even in the absence of sucrose. At sucrose concentrations above 30% (w/v), no or very little inactivation occurred even in the presence of the lactoperoxidase system. PMID- 14527781 TI - The potential of vancomycin-resistant enterococci to persist in fermented and pasteurised meat products. AB - Experiments with 148 isolates of vancomycin-resistant enterococci (VRE) were performed to assess their potential to persist and grow in fermented sausages and pasteurised meat products. All strains were meat isolates and Van-type A, except a single VanC1 strain. In total, 143 strains of Enterococcus faecium were involved. Eight selected strains were examined for their potential to grow at high salt and nitrite levels and at reduced pH. The same isolates were used in experiments with fermented sausages. All available strains were subjected to heating tests in meat suspensions with added curing ingredients. All but one of the eight tested isolates grew at pH 4.0 in tryptone soya broth (TSB). With the combination of 8% w/w NaCl, 400 ppm NaNO2 and 0.5% w/w glucose in the meat suspension, all isolates grew at 37 degrees C, whereas none grew at 7 degrees C even after 56 days. With the addition of 10% w/w NaCl, 200 ppm NaNO2 and 0.5% w/w glucose, still one E. faecium isolate grew at 37 degrees C, although very slowly. Overall, the strains tolerated high salt and nitrite concentrations and reduced pH very well, even beyond levels applied in the regular production of fermented and/or pasteurised meat products. The tested strains could be isolated after the fermentation and further ripening of "boerenmetworst" and "snijworst". Overall, their colony counts decreased on average about 1 log-unit over a period of 60 days after batter manufacture. All 148 isolates demonstrated a relatively weak thermal resistance compared to results for selected vancomycin-sensitive enterococci strains reported in the literature and to results collected under identical experimental conditions in this laboratory. None of the strains (log inoculation level about 5-6 ml(-1) for each isolate) could be cultured after heating at 70 degrees C for 10 min. PMID- 14527782 TI - Utilization of various starter cultures in the production of Amasi, a Zimbabwean naturally fermented raw milk product. AB - Fermented milk was prepared from unpasteurised milk using natural fermentation (R), back-slopping (B) and by addition of two different starter cultures (C1 and DL). The numbers of Escherichia coli, coliforms, lactic acid bacteria (LAB) and the changes in pH, carbohydrates, organic acids and volatile compounds were recorded during 48-h fermentation. After 48-h fermentation, the highest numbers of E. coli were found in R and B fermentations and the lowest in the DL fermentation. The DL culture reduced the pH faster than the other starter cultures. The DL and C1 had higher levels of LAB in the beginning of the fermentation than the other two. Galactose and lactic acid increased fastest in the DL and C1 fermentation, and R was slowest. The highest levels of succinate, ethanol and malty compounds were found in the R and B fermentations. Lower levels of LAB in the first part of the fermentations, but higher number of E. coli could explain the increased levels of succinate, ethanol and malty compounds. PMID- 14527783 TI - Effect of sodium chloride on growth and bacteriocin production by Lactobacillus amylovorus DCE 471. AB - A kinetic investigation of the effect of sodium chloride on cell growth of Lactobacillus amylovorus DCE 471 and amylovorin L471 production was carried out through in vitro experiments using a temperature and pH prevailing during sourdough fermentations. Sodium chloride interfered both with cell growth and bacteriocin production. Biomass formation and amylovorin L471 production decreased in the presence of increasing salt concentrations. Maximum bacteriocin activities were observed after the addition of 10 g l(-1) of NaCl, while the maximum specific growth rate reached an optimum at 5 g l(-1) of NaCl. High salt concentrations (20-40 g l(-1)) resulted in biphasic fermentation profiles. Based on these results, incorporation of 5-10 g l(-1) of sodium chloride in the water phase of type II sourdough preparations might be beneficial to enhance bacterial growth and amylovorin L471 production, and so contribute to the competitiveness of the strain in a sourdough environment. PMID- 14527785 TI - Behaviour of log-phase Escherichia coli at temperatures near the minimum for growth. AB - The behaviour of cold-adapted, log-phase Escherichia coli in broth cultures incubated at temperatures between 7 and 15 degrees C was examined by determinations of numbers of colonies recovered on plate count agar (PCA); absorbance at 600 nm (A600); cell lengths from photomicrographs; and cell size distributions by flow cytometry. Cultures incubated between 7 and 10 degrees C were evaluated for 8 days or until A600 values approached 1.0. Cultures incubated at > or =12 degrees C were subcultured to maintain them in the log phase for up to 8 days. Numbers of colonies recovered declined when cultures were incubated at 7 degrees C, but increased when cultures were incubated at higher temperatures. However, A600 values increased during incubation at all temperatures. The mean lengths of cells doubled during incubation at 7 degrees C for 8 days, but remained constant during incubation at 10 degrees C for 1.25 days. Forward angle light scatter (FALS) measurements obtained by flow cytometry indicated that the mean length of cells increased at < or = 8 degrees C, but not at 10 degrees C. A reference value at the 90th percentile of FALS measurements on day 0 was used to determine changes in the distribution of the lengths of cells. About 80% or 17% of the cells were above the reference value after 5 days of incubation at 7 degrees C or 1.25 days of incubation at 10 degrees C, respectively. Cultures that were maintained in the log phase at 12 degrees C became increasingly heterogeneous in cell size after 2 days, but cultures that were maintained at 13 degrees C remained constant in cell size for 8 days. The observations have implications for the prediction of mesophile proliferation at temperatures that approach their minima for growth. PMID- 14527784 TI - Biogenic amines in dry fermented sausages: a review. AB - Biogenic amines are compounds commonly present in living organisms in which they are responsible for many essential functions. They can be naturally present in many foods such as fruits and vegetables, meat, fish, chocolate and milk, but they can also be produced in high amounts by microorganisms through the activity of amino acid decarboxylases. Excessive consumption of these amines can be of health concern because their not equilibrate assumption in human organism can generate different degrees of diseases determined by their action on nervous, gastric and intestinal systems and blood pressure. High microbial counts, which characterise fermented foods, often unavoidably lead to considerable accumulation of biogenic amines, especially tyramine, 2-phenylethylamine, tryptamine, cadaverine, putrescine and histamine. However, great fluctuations of amine content are reported in the same type of product. These differences depend on many variables: the quali-quantitative composition of microbial microflora, the chemico-physical variables, the hygienic procedure adopted during production, and the availability of precursors. Dry fermented sausages are worldwide diffused fermented meat products that can be a source of biogenic amines. Even in the absence of specific rules and regulations regarding the presence of these compounds in sausages and other fermented products, an increasing attention is given to biogenic amines, especially in relation to the higher number of consumers with enhanced sensitivity to biogenic amines determined by the inhibition of the action of amino oxidases, the enzymes involved in the detoxification of these substances. The aim of this paper is to give an overview on the presence of these compounds in dry fermented sausages and to discuss the most important factors influencing their accumulation. These include process and implicit factors as well as the role of starter and nonstarter microflora growing in the different steps of sausage production. Moreover, the role of microorganisms with amino oxidase activity as starter cultures to control or reduce the accumulation of biogenic amines during ripening and storage of sausages is discussed. PMID- 14527786 TI - Black Aspergillus species as ochratoxin A producers in Portuguese wine grapes. AB - To evaluate the incidence of fungi producing ochratoxin A (OA) in Portuguese wine grapes, a survey was conducted in 11 vineyards, from four winemaking regions each with distinct climatic conditions. From setting to the harvesting period, a total of 1,650 berries were sampled by plating methods. Out of 370 aspergilli and 301 Penicillium strains isolated, 14% of the aspergilli were OA-producing strains. None of the penicillia were OA-producing strains. The black aspergilli were predominant (90%). All Aspergillus strains were tested in vitro for OA production and all were preserved in the Micoteca da Universidade do Minho (MUM) culture collection. Most of the Aspergillus carbonarius (97%) and 4% of the Aspergillus niger aggregate strains were OA producers. Almost all ochratoxigenic strains were isolated at harvest time, mainly in the regions with a Mediterranean climate. In the vineyards sampled, the percentage of colonized berries with ochratoxigenic strains was up to 38%. The vineyards from the region with Atlantic influences, with high rainfall, exhibited the lowest occurrence of Aspergillus and ochratoxigenic strains, 0% to 10% and 0% to 2% colonized berries, respectively. Data obtained here supports the hypothesis that A. carbonarius and occasionally A. niger, are the main producers of OA in grapes. In this study, the highest incidence of these fungi occurred in vineyards with a Mediterranean climate. PMID- 14527787 TI - Spoilage fungi and their mycotoxins in commercially marketed chestnuts. AB - A nationwide survey was carried out to assess mould spoilage of Castanea sativa nuts sold in Canadian grocery stores in 1998-99. Morphological and cultural characters, along with secondary metabolite profiles derived from thin-layer chromatography, were used to sort and identify fungi cultured from nut tissue. Three mycotoxigenic fungi dominated (Penicillium crustosum, Penicillium glabrum/spinulosum and Penicillium discolor) and were isolated at frequencies of 67.1%, 18.6% and 17.7%, respectively, from a total sample size of 350 nuts. Another mycotoxin producer, Aspergillus ochraceus was also isolated, but at a much lower frequency. HPLC and diode array detection were used to confirm the suspected presence of the mycotoxins penitrem A, chaetoglobosin A and C, emodin and ochratoxin A in extracts prepared from naturally infected nut tissue. To the best of our knowledge, this is the first time emodin has been found in a naturally contaminated food source. PMID- 14527788 TI - Variability of aflatoxin and cyclopiazonic acid production by Aspergillus section flavi from different substrates in Argentina. AB - Aspergillus section flavi strains isolated from peanuts, wheat and soybean grown in Argentina were screened for aflatoxins (type B and G) and cyclopiazonic acid (CPA) production. Aspergillus flavus was the predominant species in all substrates, although there was almost the same proportion of A. flavus and Aspergillus parasiticus in peanuts. Aspergillus nomius was not found. Incidence of aflatoxigenic A. flavus strains was higher in peanuts (69%) than in wheat (13%) or soybeans (5%) while the ratio of CPA producers A. flavus isolated from all substrates was very high (94% in peanuts, 93% in wheat and 73% in soybeans). Isolates of A. flavus able to produce simultaneously aflatoxins type B and CPA were detected in all substrates, suggesting the possibility of co-occurrence of these toxins. Almost all isolates of A. parasiticus resulted aflatoxins (type B and G) producers but did not produce CPA. Five of sixty-seven strains isolated from peanuts showed an unusual pattern of mycotoxin production (aflatoxins type B and G simultaneously with CPA). These strains also produced numerous small sclerotia like S strains of A. flavus detected in cottonseed in Arizona and in soils of Thailand and West Africa. The atypical strains are not widely distributed in Argentina and were found uniquely in peanuts. PMID- 14527789 TI - Viability of Saccharomyces cerevisiae cells exposed to low-amperage electrolysis as assessed by staining procedure and ATP content. AB - Assessment of yeast viability by plate counts, ATP determination and FUN-1 viability staining was performed to study sublethal injury of Saccharomyces cerevisiae cells submitted to low-amperage electrolysis. Lethal effects of electrolysis were confirmed by all methods, demonstrated by the decrease in viable counts observed during electrolysis. FUN-1 viability staining and ATP determination appeared to demonstrate higher survivors than plate counts. To study possible recovery of certain yeast cells damaged by electrolysis thus rendering them nonculturable, yeast suspensions were stored in phosphate buffer at 4 and 20 degrees C. Increase in viable counts and ATP content of treated yeast cells was observed during storage at 20 degrees C, whereas viable counts of treated and control yeast cells were shown to decrease during storage at 4 degrees C. The increase in the number of viable cells appeared to be the result of repair of damaged cells rather than regrowth of few cells remaining culturable. The lethal efficacy of electrolysis might be overestimated by plate counts. Further experiments must be done to evaluate the lethal efficacy of electrolysis on microorganisms in real conditions encountered in food products. PMID- 14527790 TI - Detection of Salmonella spp. in tropical seafood by polymerase chain reaction. AB - The incidence of Salmonella spp. in tropical seafood was studied using standard microbiological techniques and polymerase chain reaction (PCR). Six of 20 finfish (30%), 4 of 20 clams (20%) and 1 of 20 shrimp (5%) were positive by culture techniques and by PCR. In a comparative study of different selective enrichment broths and selective plating media, more than one enrichment broth and selective agar were found to be necessary for efficient detection of Salmonella from seafood. Selenite cystine broth (SCB) was found to be more efficient compared to tetrathionate broth (TTB) while both bismuth sulfite agar (BSA) and hektoen enteric agar (HEA) were equally effective as selective plating media for fish. In the case of clams, HEA was found to be more effective. The presence of Salmonella spp. could be detected by PCR amplification of DNA extracted directly from the enrichment broths. In two cases, enrichment broths that were positive by PCR did not yield Salmonella by conventional methods. PMID- 14527793 TI - Inactivation of microbes using ultrasound: a review. AB - Alternative methods for pasteurization and sterilization are gaining importance, due to increased consumer demand for new methods of food processing that have a reduced impact on nutritional content and overall food quality. Ultrasound processing or sonication is one of the alternative technologies that has shown promise in the food industry. Sonication alone is not very effective in killing bacteria in food; however, the use of ultrasound coupled with pressure and/or heat is promising. Thermosonic (heat plus sonication), manosonic (pressure plus sonication), and manothermosonic (heat and pressure plus sonication) treatments are likely the best methods to inactivate microbes, as they are more energy efficient and effective in killing microorganisms. Ultrasonic processing is still in its infancy and requires a great deal of future research in order to develop the technology on an industrial scale, and to more fully elucidate the effect of ultrasound on the properties of foods. PMID- 14527794 TI - Dynamic computer simulation of Clostridium perfringens growth in cooked ground beef. AB - The objective of this study was to develop a computer simulation algorithm to dynamically estimate and predict the growth of Clostridium perfringens in cooked ground beef. The computational algorithm was based on the implicit form of the Gompertz model, the growth kinetics of C. perfringens in cooked ground beef, and the fourth-order Runge-Kutta numerical method. This algorithm was validated using a cocktail of three strains of C. perfringens spores grown under isothermal, square-waved, linear cooling, and exponential cooling temperature profiles. In general, the results of computer simulation matched closely with the experimental data with the absolute errors less than 0.5 log(10) CFU/g. This method may be a useful tool for the food industry, regulatory agencies, distributors, and retailers to predict the effect of temperature abuse on the microbial safety of C. perfringens and other foodborne pathogens in processed meat products. PMID- 14527795 TI - Evaluation of high hydrostatic pressure sensitivity of Staphylococcus aureus and Escherichia coli O157:H7 by differential scanning calorimetry. AB - Differential scanning calorimetry (DSC) was used to evaluate the relative high hydrostatic pressure (HHP) resistances of bacterial strains from Staphylococcus aureus and Escherichia coli O157:H7 in vivo. The total apparent enthalpy change and thermal stability were two DSC parameters used to compare bacterial strains of untreated control and pressure-treated bacteria. DSC thermograms indicated that ribosomal denaturation appears to be a major factor in cell death by both thermal and high pressure treatments. However, the analysis of calorimetric data for control samples as well as pressure-treated samples clearly showed that the sensitivities of bacteria to various physical stresses can be different. While S. aureus 765 had a relatively higher resistance to thermal treatment in comparison to S. aureus 485, S. aureus 485 was determined to be more resistant to pressure than S. aureus 765. This information can be utilized in the design of processes specific to targeting certain cellular components by using different physical stresses. PMID- 14527796 TI - The microbial ecology of processing equipment in different fish industries analysis of the microflora during processing and following cleaning and disinfection. AB - The microflora adhering to the processing equipment during production and after cleaning and disinfecting procedures was identified in four different processing plants. A total of 1009 microorganisms was isolated from various-agar plates and identified. A stepwise procedure using simple phenotypic tests was used to identify the isolates and proved a fast way to group a large collection of microorganisms. Pseudomonas, Neisseriaceae, Enterobactericeae, Coryneform, Acinetobacter and lactic acid bacteria dominated the microflora of cold-smoked salmon plants, whereas the microflora in a plant processing semi-preserved herring consisted of Pseudomonas, Alcaligenes and Enterobactericeae. Psychrobacter, Staphylococcus and yeasts were found in a caviar processing plant. Overall, many microorganisms that are often isolated from fish were also isolated from the fish processing plants. However, some selection depending on processing parameters occurred, since halo- and osmo-tolerant organisms dominated in the caviar processing. After cleaning and disinfection, yeasts, Pseudomonas, Neisseriaceae and Alcaligenes remained in smokehouses, yeasts and Pseudomonas in the herring plant and Pseudomonas, Staphylococcus and yeasts in the caviar plant. The dominant adhering organisms after cleaning and disinfection were pseudomonads and yeasts independently of the microflora during processing. Knowledge of the adhering microflora is essential in the Good Hygienic Practises programme of food processing plants, as the development and design of improved cleaning and disinfecting procedures should target the microorganisms persisting and potentially contaminating the product. PMID- 14527797 TI - Efficacy of sorbates on the control of the growth of Eurotium species in bakery products with near neutral pH. AB - The effects of sorbic acid and potassium sorbate on growth of different Eurotium isolates when added to a bakery product analogue were tested under different environmental conditions. Water activity of the products was adjusted to values in the range of 0.75-0.90, and storage temperatures were in the range of 15-30 degrees C. Preservatives were added in concentrations ranging from 0.025% to 0.2%. It was observed that 0.025% and 0.05% concentrations always enhanced the isolates growth, while 0.1% had little preservative effect. Finally, even the highest concentration (0.2%) was not suitable as it only controlled fungal growth under certain water activity and temperature levels. It was concluded that these weak-acid preservatives are not useful when added to bakery products with near to neutral pH. PMID- 14527798 TI - Phytase activity in sourdough lactic acid bacteria: purification and characterization of a phytase from Lactobacillus sanfranciscensis CB1. AB - The phytase activity of 12 species of sourdough lactic acid bacteria was screened. It was intracellular only, largely distributed among the species and strains of Lactobacillus sanfranciscensis possessed the highest levels of activity. A monomeric ca. 50-kDa phytase was purified to homogeneity from L. sanfranciscensis CB1 by three chromatographic steps. L. sanfranciscensis CB1 exhibited the highest hydrolysing activity on Na-phytate after reaching the stationary phase of growth (ca. 12 h). Cells cultivated in the presence of maltose and fructose showed an increase of the phytase activity of ca. 35% with respect to the other carbon sources used. The phytase was optimally active at pH 4.0 and 45 degrees C. The enzyme was strongly inhibited by 2 mM of phenylmethylsulfonyl fluoride (PMSF), and 2 mM Hg(2+) and Fe(2+). It had a pI of ca. 5.0. The substrate specificity was dependent on the type of phosphate ester; a very low activity was detected on alpha-D-glucose-1-phosphate and D-fructose-6- and 1,6-phosphate, while the highest hydrolysis was found towards adenosine-5' tri-, di- and mono-phosphate. Compared to these substrates, the activity on Na phytate was also relevant. The enzyme was thermo-stable after exposure to 70 degrees C for 30 min; the D value calculated at 80 degrees C was ca. 10 min. As shown by the Central Composite Design (CCD) applied to study the individual and interactive effects of pH, temperature and NaCl, acidic conditions and elevated temperatures were indispensable for the enzyme adaptation to high NaCl concentrations. L. sanfranciscensis CB1 cells or the correspondent cytoplasmic extract were used to ferment a sourdough for 8 h at 37 degrees C; a marked decreased (64-74%) of the Na-phytate concentration was found compared with the unstarted dough. The sourdough started with L. sanfranciscensis CB1 cells was re used for several times and the phytase activity was maintained to a considerable level. PMID- 14527799 TI - Comparison of the BAX System with a multiplex PCR method for simultaneous detection and identification of Campylobacter jejuni and Campylobacter coli in environmental samples. AB - The Campylobacter detection is performed by conventional culture methods and the identification of Campylobacter jejuni and Campylobacter coli is principally based on the hippurate hydrolysis test. The two major drawbacks of this biochemical test for species identification include the inconsistency of the results and the presence of atypical strains, which can lead to the misidentification of an isolate. As an alternative, multiplex polymerase chain reaction (mPCR) protocols for the simultaneous detection and identification of different Campylobacter species have been developed. This study examined the performances of an experimental BAX System assay for the C. jejuni and C. coli identification in comparison to a multiplex PCR protocol recently published. The samples tested were represented by 106 environmental swabs collected on Teflon strips and tables, stainless steel saws, hooks and trays, ceramic floors and walls, as well as equipment surfaces, located in a swine (N=50) and a poultry (N=56) slaughterhouse. The highest Campylobacter detection rate was obtained after 48 h of enrichment by using both the PCR procedures. After 24 h, the BAX System provides a more rapid and accurate Campylobacter detection and identification assay than the multiplex PCR. Except for two samples, all the broths where Campylobacter cells were detected after 24 or 48 h of enrichment, with at least one of the PCR protocols, gave Campylobacter colonies using the culture method. PMID- 14527800 TI - An evaluation of rapid methods for detecting Escherichia coli O157 on beef carcasses. AB - Numbers of Escherichia coli O157 in food may be low and sensitive techniques are therefore needed for its detection. The objectives of this study were to use carcass meat samples artificially inoculated with various strains of E. coli O157 to compare the sensitivity of enrichment in three different media and to compare immunomagnetic separation followed by culture of magnetic beads to cefixime tellurite sorbitol MacConkey agar with three immunoassays for the detection of E. coli O157 in the enrichment cultures. Duplicate 250, 25 and 2-3 CFU of each of 16 strains of E. coli O157 added to 25-g samples of beef carcass meat were used to compare the sensitivity of (1) enrichment in supplemented tryptone soya broth (sTSB), Reveal 8-h and Reveal 20-h media, and (2) immunomagnetic separation and culture to cefixime tellurite sorbitol MacConkey agar (IMS/CT-SMAC) with Reveal, VIP and STAT immunoassays for detecting the organism. An initial inoculum of 250 CFU/25 g meat was detected in all 32 samples by IMS/CT-SMAC performed on all enrichment media and by Reveal performed on Reveal 8-h and Reveal 20-h media, but in only 30, 19 and 9 samples by Reveal, VIP and STAT, respectively, performed on sTSB medium. An initial inoculum of 25 CFU/25 g meat was detected in 28, 32 and 30 of 32 samples by IMS/CT-SMAC performed on sTSB, Reveal 8-h and Reveal 20-h media, respectively, and in 32 and 30 samples by Reveal performed on Reveal 8-h and Reveal 20-h media, but in only 22, 11 and 2 samples by Reveal, VIP and STAT, respectively, performed on sTSB medium. An initial inoculum of 2-3 CFU/25 g meat was detected in 25, 31 and 28 of 32 samples by IMS/CT-SMAC performed on sTSB, Reveal 8-h and Reveal 20-h media, respectively, and in 25 and 23 samples by Reveal performed on Reveal 8-h and Reveal 20-h media, but in only 14, 1 and 0 samples by Reveal, VIP and STAT, respectively, performed on sTSB medium. PMID- 14527801 TI - Effect of potentially probiotic lactobacilli on faecal enzyme activity in minipigs on a high-fat, high-cholesterol diet-a preliminary in vivo trial. AB - Minipigs were fed a "Western-style", high-cholesterol diet for a baseline period, followed by the diet containing a mixture of three Lactobacillus strains with potential probiotic features, after which a normal pig diet was followed. The faecal enzyme activity for beta-glucuronidase and azoreductase, which are commonly considered as markers for procarcinogenic activity, was significantly reduced during the 5 weeks of "probiotic" supplementation. During the period of Lactobacillus administration, the cell counts for total anaerobes increased, whereas the total number of aerobes showed no change. PMID- 14527802 TI - Identification of proteins induced at low pH in Lactococcus lactis. AB - The Gram-positive bacterium Lactococcus lactis is of major importance to the dairy industry due to its conversion of lactose to lactic acid leading to the acidification of milk. To investigate which proteins are induced when L. lactis is exposed to conditions of low pH, we used two-dimensional gel electrophoresis to follow how protein expression changes with the degree of acidification. We found that reducing the pH of the growth medium with hydrochloric acid induced the synthesis of a small subset of proteins. The majority of these proteins were induced both after a minor (pH 5.5) and a major (pH 4.5) reduction in pH. Among the most strongly induced proteins, we identified the oxidative stress proteins superoxide dismutase and alkylhydroperoxidase as well as the autoinducer synthesis protein, LuxS. We also observed a differential induction of heat shock proteins by low pH as members of the CtsR regulon, ClpE and ClpP were induced at both pH 5.5 and 4.5, while HrcA-regulated chaperones, GroEL, GroES, DnaK and GrpE were induced only at pH 4.5. In addition, we identified two proteins repressed by low pH that proved to be the L. lactis HPr protein of the phosphoenolpyruvate sugar phosphotransferase system and the trigger factor known to participate in the folding of newly synthesized polypeptides. PMID- 14527803 TI - Efficacy of chlorine and calcinated calcium treatment of alfalfa seeds and sprouts to eliminate Salmonella. AB - The efficacy of a 20,000 ppm calcium hypochlorite treatment of alfalfa seeds artificially contaminated with Salmonella was studied. Salmonella populations reached >7.0 log on sprouts grown from seeds artificially contaminated with Salmonella and then treated with 20,000 ppm Ca(OCl)(2). The efficacy of spray application of chlorine (100 ppm) to eliminate Salmonella during germination and growth of alfalfa was assessed. Alfalfa seed artificially contaminated with Salmonella was treated at germination, on day 2 or day 4, or for the duration of the growth period. Spray application of 100 ppm chlorine at germination, day 2, or day 4 of growth was minimally effective resulting in approximately a 0.5-log decrease in population of Salmonella. Treatment on each of the 4 days of growth reduced populations of Salmonella by only 1.5 log. Combined treatment of seeds with 20,000 ppm Ca(OCl)(2) and followed by 100 ppm chlorine or calcinated calcium during germination and sprout growth did not eliminate Salmonella. PMID- 14527804 TI - Repeated spinal flexion modulates the flexion-relaxation phenomenon. AB - OBJECTIVES: To determine if repeated spinal flexion and loading modulate the deactivation of lumbar muscles near full flexion (flexion-relaxation). DESIGN: Repeated measures experimental study of the effect of repetitive trunk flexion and added mass on the flexion-relaxation phenomenon. BACKGROUND: Repeated flexion causes muscular fatigue, creep of passive tissues and diminished protective reflexes. However, flexion-relaxation has not been studied in repeated trunk flexion, and could be related to the increased risk of low-back disorders. METHODS: Thirty healthy young subjects performed 100 trunk flexion movements between standing and full flexion. Erector spinae electromyography and lumbar spine flexion were measured during cycles 1-10 (no load), 11-20 (performed holding a mass in the hands), 81-90 (mass in the hands) and 91-100 (no load). The spinal flexion angle at myoelectric silence and full flexion were extracted from each movement cycle. RESULTS: Twenty-three of the 30 subjects showed flexion relaxation throughout the repeated trunk flexion. The flexion-relaxation and maximum flexion angles increased at the end of the experiment; the flexion relaxation angle relative to the maximum flexion angle also increased. This effect depended on the load condition; the flexion-relaxation and maximum flexion angles showed a greater increase in the unloaded than loaded condition. CONCLUSIONS: The flexion-relaxation phenomenon was changed due to repeated trunk flexion. The increases in flexion-relaxation angle likely involve changes to the neuromuscular control system. RELEVANCE: The deactivation of the erector muscles near full flexion occurs at a greater spinal flexion angle and a greater proportion of maximum spinal flexion following repeated spinal flexion. This may be related to the increased risk of injury associated with repeated flexion. PMID- 14527805 TI - Biomechanical responses of the intervertebral joints to static and vibrational loading: a finite element study. AB - OBJECTIVE: This study was performed to investigate the time-dependent responses of the intervertebral joint to static and vibrational loads. DESIGN: A poroelastic finite element model was established to analyse the fluid flow, stress distribution and deformation of the intervertebral disc. BACKGROUND: Long term exposure to whole body vibration is highly associated with disc degeneration and low back pain. It is hypothesized that moderate vibrational loading may increase the efficiency of fluid and nutritional transport of the intervertebral disc while prolonged static and excessive vibrational loading may have deleterious effect. METHODS: A three-dimensional finite element model was established using the actual geometry of the L4-L5 lumber motion segment. Nonlinear poroelastic properties were assigned to the intervertebral disc and cancellous bone. Static and vibrational loads were applied and the responses of fluid flow and stress distributions were analysed. RESULTS: The finite element model showed that the loads carried by the annulus and the facets increased with time under static loading. The fluid flow and deformation of the intervertebral disc were dependent on the loading frequency. CONCLUSION: Vibration loading may be able to enhance disc fluid exchange via the fluid pumping mechanism. RELEVANCE: The predicted responses implied that vibrational motion may be important in facilitating fluid and metabolic transport of the intervertebral disc. PMID- 14527806 TI - Estimation of spinal loading in vertical vibrations by numerical simulation. AB - OBJECTIVE: This paper describes the prediction of spinal forces in car occupants during vertical vibrations using a numerical multi-body occupant model. BACKGROUND: An increasing part of the population is exposed to whole body vibrations in vehicles. In literature, vertical vibrations and low back pain are often related to each other. The cause of these low back pains is not well understood. A numerical human model, predicting intervertebral forces, can help to understand the mechanics of the human spine during vertical vibrations. METHODS: Numerical human and seat models have been used. Human model responses have been validated for vertical vibrations (rigid and standard car seat condition): simulated and experimental seat-to-human frequency response functions have been compared. The spinal shear and compressive forces have been investigated with the model. RESULTS: The human model seat-to-pelvis and seat-to T1 frequency response functions in the rigid seat condition and all seat-to-human frequency response functions in the standard car seat condition approach the experimental results reasonably. The lumbar and the lower thoracic spine are subjected to the largest shear and compressive forces. CONCLUSIONS: The human model responses correlate reasonable with the volunteer responses. The predicted spinal forces could be used as a basis for derivation of hypothetical mechanisms and better understanding of low back pain disorders. RELEVANCE: In order to solve the problem of whole body vibration related injuries, knowledge about the interaction between human spinal vertebrae in vertical vibrations is required. This interaction cannot be measured in volunteer experiments. This paper describes the application of a numerical human model for prediction of spinal forces, that could be used as a basis for derivation of hypotheses regarding low back pain disorders. PMID- 14527807 TI - The responses of leg and trunk muscles to sudden unloading of the hands: implications for balance and spine stability. AB - OBJECTIVE: To examine the anticipatory and responsive actions of leg and trunk muscles, and their role in whole-body and spine control in situations of sudden unloading of the hands in the sagittal plane. DESIGN: EMG and force plate measures were used to determine the baseline, anticipatory responses and post unloading responses of selected trunk and leg muscles under different conditions of unload timing knowledge. BACKGROUND: Postural muscles have been observed to increase activation in anticipation of a known loading situation to decrease the overall effect of an impulsive load delivered to the spine. It is thought that this increased activation places the spine in a more stable state, thereby reducing the likelihood of injury. Comparisons have not been made previously of the responses of postural muscles to unloading conditions where the certainty of unload timing is varied. METHODS: Eleven male subjects, holding a 6.8 kg load in the hands, were subjected to three different unloading conditions: (1) voluntary load drop; (2) known timing of load release; (3) unknown timing of load release. Anterior-posterior center of pressure data, as well as EMG activity on 8 right side muscles, were collected for 10 trials in each condition. RESULTS: Anterior posterior center of pressure responses were significantly different (P<0.05) between each of the three conditions. Lumbar erector spinae and thoracic erector spinae significantly decreased anticipatory activity as knowledge of the unload timing increased. Five of the eight monitored muscles demonstrated significantly decreased response levels as knowledge of the timing of unloading increased. CONCLUSIONS: When an unload is self-triggered, preparatory adjustments can be made which reduce the overall postural perturbation to the body, and the spine in particular, while minimizing the responsive activity of trunk muscles. RELEVANCE: Spinal instability has been identified as a risk factor for low back injury during trunk loading. This study demonstrates that, in situations of sudden unloading, knowledge of the timing of the unloading may lead to anticipatory actions of postural muscles which actually decrease spinal stability, thereby increasing the risk of injury were an unexpected perturbation to occur. PMID- 14527808 TI - Force and time recording for validating the sacroiliac distraction test. AB - OBJECTIVE: The present aim was to investigate whether applied force and time interval of force exposure during the sacroiliac distraction test could discriminate subjects with sacroiliitis pain from healthy subjects. DESIGN: The study was a methodological one using force plates sensing three orthogonal forces. BACKGROUND: Studies evaluating the validity of sacroiliac joint tests show conflicting results. To our knowledge, no previous studies have evaluated force recording combined with pain reports. METHODS: Eleven subjects with sacroiliitis and 11 healthy subjects were examined with distraction test once by each of three physiotherapists. The subjects indicated any pain by pressing a button giving a mark in the data collection. RESULTS: The magnitude of the total force vector was significantly smaller (P<0.05) and the time interval was significantly shorter (P<0.05) in the sacroiliitis group, although almost 20 s could elapse before pain was provoked. The sensitivity of the test was 0.55 calculated for all physiotherapists involved, and varied between them, range 0.55 0.82. The specificity was 1.0. CONCLUSIONS: The results support the advantage of force and time recording during the distraction test as a technique for evaluating pain. Further investigation is needed on whether the velocity of force application, the ability to maintain the same pressure and the bilateral distribution of force on the sacroiliac joints influence the sensitivity. RELEVANCE: There is a need for objective evaluation methods in manual tests that are easy to perform. Recording force and its time interval may be a step towards standardising pain-provocation sacroiliac joint tests. PMID- 14527809 TI - Global 3D head-trunk kinematics during cervical spine manipulation at different levels. AB - OBJECTIVE: Determination of the three-dimensional kinematics of the head relative to the upper trunk obtained during a manipulation applied on two different cervical levels and on both sides. DESIGN: Descriptive study performed on 14 asymptomatic volunteers. The range of motion was measured by a 3D electrogoniometer during manipulation executed by the same practitioner. BACKGROUND: Spinal manipulative therapy is a common treatment approach in patients suffering from some spinal disorders. Complications exist; they are thought to be related to the force applied by the practitioner and the range of spinal motion obtained during the manipulation. Yet, little is known about cervical spine motion during manipulation. METHODS: Three dimensional electrogoniometry using a 6 degree-of-freedom spatial linkage fixed between the head and the upper trunk was used to record the pattern of motion and the amplitudes obtained during a manipulation on two cervical levels (C3 and C5) and on left and right sides. On single practitioner applied the same technique to all subjects in a seated position. RESULTS: The side and the spinal level manipulated did not influence 3D ranges of motion. The mean ranges of motion obtained were 30 degrees axial rotation, 46 degrees lateral bending and 2 degrees flexion. A significant difference of the flexion-extension range existed between manipulations with and without audible release. Axial rotation and lateral bending ranges were correlated. Except for lateral bending which was close to active range, the motion ranges obtained during manipulation were well below active range of motion reported in literature. CONCLUSIONS: The results of this study suggest that for the kind of manipulation applied, maximal amplitude between head and trunk does not exceed physiological active range of motion. The amplitude for rotation, which is generally assumed to involve greatest risks for negative side effects, is significantly lower than during active motion. As the study was performed with one practitioner, this result may only be generalized with care. RELEVANCE: In spinal manipulative therapy, extreme range of motion as the result of the forces applied is generally believed to represent a major risk for negative side effects, especially with regard to the cervical spine. With a multiple component technique, amplitudes between head and upper trunk were shown not to differ significantly with regard to the side nor to the spinal level. Recorded ranges of motion did not exceed those reported for active motion in literature. PMID- 14527810 TI - Postural adjustments to voluntary head motions during standing are modified following stroke. AB - OBJECTIVE: To compare postural adjustments to voluntary head motions during standing between stroke and healthy subjects. DESIGN: Quasi-experimental design involving two groups: stroke patients (n=8) and age-matched healthy controls (n=5). BACKGROUND: Massive sensory stimulation from the visual, vestibular and neck proprioceptive systems is conveyed during a voluntary head motion. It remains to be investigated how stroke patients integrate this sensory information and generate appropriate postural adjustments to maintain standing balance. METHODS: Eight stroke patients and five healthy age-matched subjects performed fast head motions (up, down, right, left, or none) in random sequence during standing. Body kinematics in 3-D, horizontal displacement of the center of pressure and the body center of mass, as well as bilateral vertical ground reaction forces and activation of tibialis anterior, medial gastrocnemius, vastus lateralis and semitendinosus muscles, were acquired and analyzed. RESULTS: Stroke patients moved their heads at lower velocities while maintaining similar range of motion as compared to healthy subjects. Stroke patients showed larger difference in center of pressure and center of mass excursions and larger root-mean-square values for center of pressure and center of mass displacement and velocity during all directions of head motions. Stroke patients presented disrupted loading patterns to avoid weight bearing of the paretic limb, in contrast to the typical and direction-specific loading patterns observed in healthy subjects during head motions. CONCLUSIONS: Stroke patients manifest altered postural adjustments to voluntary head motions during standing. It suggests that sensorimotor integration processes for stance and balance are impaired by a stroke. RELEVANCE: Head motions are expected to produce little displacement in body center of mass, as the mass of the head is relatively small as compared to the rest of the body. However, the complex information channeled from vestibular, visual and neck proprioceptive afferents must be processed by the central nervous system in relation to the motor commands, such that head motions can be executed smoothly and rapidly while maintaining standing balance. The altered postural adjustments to head motions observed in stroke patients suggest an underlying impairment in sensorimotor integration. PMID- 14527811 TI - Evaluation of shoulder proprioception following muscle fatigue. AB - OBJECTIVE: To investigate the effect of shoulder muscle fatigue on glenohumeral proprioception. DESIGN: A repeated proprioception test was performed. BACKGROUND: The role of conditioning and fatigue in sport injuries remains controversial. It has been hypothesized that proprioceptive information plays an important role in joint stabilization and that muscle fatigue may alter proprioceptive ability. However, the effect of shoulder muscle fatigue on glenohumeral proprioception is still controversial. METHODS: Eleven normal subjects (mean age 27.3 years) participated in this study. Proprioception tests (on the dominant shoulder) were performed in which proprioception of the active reproduced and passive reproduced shoulder position was measured using an isokinetic dynamometer and a proprioception testing apparatus, respectively. The speed of active repositioning was at 2 deg/s and passive repositioning was at 0.5 deg/s. The mean value of maximum voluntary contraction and the number of repetitions for muscle fatigue were recorded. Post-fatigue proprioception test was started within 3 min after muscle fatigue. RESULTS: There was no significant difference of shoulder proprioception between pre- and post-fatigue determinations of passive repositioning in shoulder internal rotation, passive repositioning in external rotation and active repositioning in internal rotation. There was a significant difference between pre- and post-fatigue determination of active repositioning in external shoulder rotation (mean degrees: 2.57 (SD 1.02) vs. 4.96 (SD 1.73), P<0.05). CONCLUSION: Shoulder proprioception in active repositioning in external rotation was major affected by muscle mechanoreceptors in the presence of muscle fatigue. RELEVANCE: This study revealed that the shoulder proprioception after muscle fatigue in active repositioning in shoulder external rotation was affected more sensitively by the muscle mechanoreceptors than the joint mechanoreceptors. Increasing resistance of muscle fatigue would increase the shoulder proprioceptive ability. PMID- 14527812 TI - Joint torques and dynamic joint stiffness in elderly and young men during stepping down. AB - OBJECTIVE: To compare the joint torque pattern and dynamic joint stiffness at the knee and ankle in elderly and young men during stepping down. BACKGROUND: Adequate joint stiffness is critical during the single support phase to control forward and downward body momentum. DESIGN: Six active elderly men (mean 67.7) and six young men (mean 23.6) of similar body mass and height, were filmed stepping down from one force platform to another. Repeated trials were undertaken at three different step heights (200, 250, and 300 mm). METHOD: Joint torques were determined for the ankle and knee of the support limb throughout the single support phase. The gradient of the joint torque-angle graph was calculated to define dynamic joint stiffness of the ankle and knee in two phases; (I) from initiation of movement until heel-off of the supporting limb, and (II) from heel off of the supporting limb to contra-limb touch down. RESULTS: Maximum ankle torque values were lower in the elderly and occurred at a larger dorsiflexion angle (P<0.05). Knee torque patterns were similar in both groups. Phase I ankle stiffness was significantly less in the elderly (4.0-5.2 Nm/ degrees ) at all step heights compared to the young (7.6 - 8.7 Nm/ degrees ). In both groups ankle stiffness in Phase II increased with step height, while knee joint stiffness decreased. CONCLUSIONS: The different torque pattern and lower dynamic ankle stiffness in the elderly, particularly for Phase I, suggested an altered control strategy. These findings highlight the importance of dynamic ankle joint stiffness in stepping down. RELEVANCE: Understanding how the elderly step down may be important in developing strategies to prevent falls. PMID- 14527813 TI - Fixation of trochanteric slide osteotomies: a biomechanical study. AB - OBJECTIVE: (1) Determine the effect of a compressive force on the stability of trochanteric slide osteotomies repaired with a cable repair system or a suturing technique. (2) Develop an approach to surgical decision making for trochanteric repair. DESIGN: Muscle forces acting on the greater trochanter were experimentally modeled by the application of shear and compressive loads to osteotomized greater trochanters. A repeated measures design was used to compare suture and cable fixation. BACKGROUND: The use of cables and wires for trochanteric repair has been associated with a high incidence of acetabular loosening and trochanteric bursitis. With trochanteric slide osteotomies, the vastus lateralis remains attached to the trochanter, which results in a compressive force being generated across the osteotomy and relatively small shear forces. The use of less rigid fixation techniques for trochanteric repair, such as sutures, may reduce the complications of cables and wires. METHODS: Seven cadaveric femora with trochanteric osteotomies were tested sequentially after repair with a cable system and with a suturing technique. A cyclic shear load of constant amplitude was applied while a compressive load was decreased in a stepwise fashion. Migration and cyclic motion of the trochanter were measured, and the coefficient of friction was also determined. RESULTS: Cyclic motions of the trochanter in both superior and anterior directions were generally less than 0.5 mm and were not significantly different between the cables and sutures at high compressive loads. At low compressive loads, cyclic motion was significantly lower with the cable system. CONCLUSIONS: Compression across the trochanteric slide osteotomy has a significant effect on stability. Cyclic motion of the trochanter is similar for both suture or cable repair of a trochanteric slide with good preservation of soft tissue attachments. RELEVANCE: Based on theoretical and experimental evidence, repair of trochanteric slide osteotomies with a suture technique may be a viable alternative to the use of cables and wires in selected cases. PMID- 14527814 TI - The strength of different fixation techniques for bicondylar tibial plateau fractures--a biomechanical study. AB - OBJECTIVE: To compare the mechanical strength of fixation of bicondylar tibial plateau fractures using internal and external fixation techniques. DESIGN: A randomised laboratory testing of fixation methods currently in clinical use. BACKGROUND: To reduce the high complication rate associated with the double plating, many surgeons have started to use less invasive techniques. However the literature comparing the mechanical performance of these methods is still scarce. METHODS: Bicondylar tibial plateau fractures were simulated on Sawbones tibiae and fixed with one of the five fixation methods: dual plating, a two-ring hybrid fixator with inter-fragmentary screws, a ring-bar hybrid fixator with inter fragmentary screws, lateral plate and medial monolateral external fixator, lateral plate and medial inter-fragmentary screws. Following statistical power calculations, seven tibiae were used for each fixation method. The specimens were tested in cyclical compression to failure. The vertical subsidence in the medial or lateral plateau was measured using extensometers. RESULTS: In all cases the mode of failure was consistent with collapse occurring in the medial plateau. There was no significant difference in the failure load between dual plating and the two-ring hybrid fixator (4218 N and 4184 N respectively; P=0.28, t-test). Failure was seen at significantly lower loads with the other fixation systems. CONCLUSION: Weight-bearing mobilisation of the patient may be undertaken earlier with more confidence by using the double plating or two-ring hybrid fixator rather than other less strong techniques. The choice on which of these two methods to use may depend on tissue viability and surgeon preference. PMID- 14527815 TI - Muscle activation pattern and gait biomechanics after total knee replacement. AB - OBJECTIVE: The aim of this study is to evaluate residual muscle function abnormalities after total knee replacement, with respect to gait kinematics and kinetics. DESIGN: Longitudinal study on a follow-up of up to two years. BACKGROUND: Gait usually presents an excellent improvement after total knee replacement. Nevertheless, some kinematics and kinetics abnormalities persist even after a long period of time and they might have implications in long-term prosthesis failure. Additionally, lower limb muscle activity has not been sufficiently studied in the past directly by means of dynamic EMG. METHODS: Nine patients who had a posterior cruciate sparing total knee replacement design were evaluated by means of clinical assessment and gait analysis at the end of rehabilitation trials at six, twelve and twenty four months. EMG from trunk and lower limb muscles was registered and elaborated through a statistical detector for the on-off timing. RESULTS: Gait analysis showed a slow gait, with a "stiff knee gait pattern" and prolonged muscular co-contractions during stance. CONCLUSIONS: Knee kinematics and kinetics abnormalities during loading acceptance after total knee replacement are associated with co-contractions in muscular activation pattern. RELEVANCE: Gait pattern at two years after total knee replacement is deemed unphysiological, potentially dangerous for the implant duration in time and responsible for residual disability in patients. Muscular behavior during daily activities provides insight into the integration of the prosthetic biomechanics within the muscular-skeletal system. This information is of relevance to improve prosthetic design, rehabilitation programs and knee motor performance. PMID- 14527816 TI - Aligning anatomical structure from spiral X-ray computed tomography with plantar pressure data. AB - OBJECTIVE: To determine the reliability of identifying the location of each metatarsal head from spiral X-ray computed tomography on a matrix type pressure sensor recording. DESIGN: Experimental; test-retest and within subject two condition comparison. BACKGROUND: Plantar pressure data quantify the amount of pressure and the general location of pressure. Specific anatomical structures associated with the site of pressure can only be determined by registering radiological data to the pressure sensor. METHODS: Eighteen subjects, nine individuals without diabetes mellitus and nine individuals with diabetes mellitus, peripheral neuropathy, and a history of plantar foot ulcers, participated in plantar pressure testing and spiral X-ray computed tomography scanning of their foot. A registration technique was developed to align spiral X ray computed tomography data with pressure sensor data. RESULTS: When mapping the metatarsal head locations to the pressure sensor 48 of 90 metatarsal head locations were identical between repeated test occasions (53.3%), 40 of 90 metatarsal head locations were one pixel (5.08 mm) different between repeated test occasions (44.4%), and 2 of 90 were two pixels different (2.2%). Ninety eight percent of repeated measures replicated exactly or varied by one pixel. CONCLUSION: Anatomical data from spiral X-ray computed tomography scanning can be reliably co-registered with pressure data. RELEVANCE: The ability to match bony anatomy to pressure data allows better understanding of structural factors contributing to peak pressure, provides precise information for three-dimensional modeling of the foot, and can improve orthotic fabrication and modification aimed at reducing pressure on the bottom of the foot. PMID- 14527817 TI - Antihyperlipidemic effects of different molecular weight sulfated polysaccharides from Ulva pertusa (Chlorophyta). AB - Ulvan, a sulfated polysaccharide from Ulva pertusa, was degraded to yield two low molecular weight fractions U1 and U2. The molecular weights of ulvan and its fractions were determined and varied from 151.6 to 28.2 kDa. They were fed to rats on a hypercholesterolemic diet for 21 days to evaluate and compare the antihyperlipidemic actions. Ulvan-based diet significantly lowered the levels of serum total cholesterol (-45.2%, P<0.05) and low density lipoprotein cholesterol (LDL-cholesterol, -54.1%, P<0.05). While U1- and U2-based diets significantly elevated the levels of serum high density lipoprotein cholesterol (HDL cholesterol, +22.0% for U1, not significant; +61.0% for U2; P<0.05) and reduced triglyceride (TG, -82.4% for U1, -77.7% for U2; P<0.05) in rats as compared to control diet. In addition, consumptions of various ulvans significantly increased fecal bile acid excrement. The results indicated that ulvans with different molecular weights exhibited diverse effects on lipid metabolism. The high molecular weight ulvan was effective in serum total and LDL-cholesterol, whereas low molecular weight fractions were in TG and HDL-cholesterol. The fractions were considered to be more beneficial to hyperlipidemia associated with diabetes over ulvan. PMID- 14527818 TI - Antidiabetic effect of Gymnema montanum leaves: effect on lipid peroxidation induced oxidative stress in experimental diabetes. AB - Gymnema montanum is widely used in ancient medicine for the ailment of various diseases. Oral administration of 200 mg kg(-1) (body weight) BW of the alcoholic extract of the leaf for 3 weeks resulted in a significant reduction in blood glucose and an increase in plasma insulin, whereas the effect of 50 and 100 mg kg(-1) BW was not significant. The alcoholic extract also resulted in decreased free radical formation in plasma of diabetic rats. Thus, this study shows that Gymnema montanum leaf extract (GLEt) possess antihyperglycemic and antiperoxidative effect. The decrease in lipid peroxides and increase in reduced glutathione (GSH), ascorbic acid (Vitamin C) and alpha-tocopherol (Vitamin E) clearly show the antioxidant properties of GLEt. The effect of GLEt was most prominently seen in the case of animals given 200 mg kg(-1) BW. In addition, the results suggest that GLEt was highly effective than the reference drug glibenclamide. PMID- 14527819 TI - Infarct size limiting effect of apstatin alone and in combination with enalapril, lisinopril and ramipril in rats with experimental myocardial infarction. AB - Bradykinin is a potent vasoactive peptide that is known to elicit a number of biological responses. A number of peptidases have been identified to possess kininase activity, the inhibition of which increases the availability and effectiveness of kinins. We wished to determine the cardioprotective actions of an aminopeptidase P inhibitor, apstatin alone and in combination with enalapril/lisinopril/ramipril in an in vivo rat model of acute ischemia (30 min) and reperfusion (4 h). Myocardial infarction was produced by occlusion of the left anterior descending coronary artery for 30 min followed by 4 h of reperfusion. Infarct size was measured by using the staining agent 2,3,5 triphenyl tetrazolium chloride (TTC). Lipid peroxide levels in serum and in heart tissue were estimated spectrophotometrically. A lead II electrocardiogram was monitored at various intervals throughout the experiment. Infarct size was reduced to a greater extent with apstatin and with combined inhibition it was further reduced. Infarct size reduction obtained with the combined inhibition came to normal with the prior administration of B2 bradykinin antagonist HOE140 suggests the involvement of bradykinin in the cardioprotective actions of apstatin. PMID- 14527820 TI - Biochemical changes on the cardioprotective effect of nicorandil and amlodipine during experimental myocardial infarction in rats. AB - The synergistic protective effect of nicorandil (KATP channel opener) and amlodipine (calcium channel blocker) on mitochondrial respiration and mitochondrial lipid contents were examined on isoproterenol-induced myocardial infarction in rats. The rats given isoproterenol (150 mg kg(-1) daily, i.p.) for 2 days showed significant changes in mitochondrial respiration and mitochondrial lipid profile levels. Pretreatment with nicorandil (2.5 mg kg(-1) daily, p.o.) and amlodipine (5.0 mg kg(-1) daily, p.o.) for 3 days significantly prevented these alterations and restored the mitochondrial respiration and mitochondrial lipid contents to near normal. Histopathological observations were also in correlation with the biochemical parameters. These findings indicate the synergistic protective effect of nicorandil and amlodipine on mitochondrial respiration and its membrane integrity during isoproterenol-induced cardiac damage. PMID- 14527821 TI - Apoptosis induced by simvastatin in rat vascular smooth muscle cell through Ca2+ calpain and caspase-3 dependent pathway. AB - Several studies have shown that simvastatin induces apoptosis in a variety of cell lines including vascular smooth muscle cells (VSMCs), but the exactly mechanisms involved in it is not very clear. The aim of this study was to investigate the mechanisms and signal pathways involved in apoptosis induced by simvastatin. When exposed to 30 microM simvastatin, [Ca2+]i in VSMCs increased with time and reached to 336 +/- 52 nM at 6 h, more than four-fold of control (P<0.01, n=5). Verapamil (80 microM), a membrane voltage-dependent Ca2+ channel blocker, attenuated simvastatin-induced augmentation of free calcium concentration from 336 +/- 52 nM to 144 +/- 34 nM (P<0.01). After being exposed to 30 microM simvastatin for 8 h, calpain activity markedly increased (P<0.05, n=4) and reached to more than three-fold of control at 12 h (P<0.01). Caspase-3 was also activated by simvastatin after 12 h. Verapamil and PD150606, a cell permeable selective calpain inhibitor, significantly inhibited simvastatin induced augmentation of calpain activity and blocked caspase-3 activation, respectively. Furthermore, 80 microM verapamil and 100 microM PD150606 decreased simvastatin-induced apoptosis rate from 24.2 +/- 1.7% to 7.9 +/- 0.6% (P<0.01, n=4) and 9.5 +/- 1.9% (P<0.01), respectively and also prevented simvastatin induced DNA laddering. In conclusion, we indicated that simvastatin increases cytosolic free calcium concentration mainly through calcium influx from extracellular solution and then induces apoptosis by activating caspase-3 via calcium-dependent protease calpain. PMID- 14527822 TI - Effect of thiamine on 3H-MPP+ uptake by Caco-2 cells. AB - Recent studies on the intestinal uptake of the organic cation 1-methyl-4 phenylpyridinium (MPP+) showed that transport of this compound occurs through human extraneuronal monoamine transporter (hEMT). Moreover, it was recently described that alkaline phosphatase (ALP), an ecto-phosphatase anchored to the plasma membrane and able to dephosphorylate extracellular substrates or cell surface proteins, is directly or indirectly involved in the modulation of MPP+ uptake by Caco-2 cells. The present study investigated a putative modulation of MPP+ intestinal apical uptake and ecto-ALP activity by thiamine (T+) and thiamine pyrophosphate (TPP, a T+ dietary precursor). For this purpose, we used Caco-2 cells, an enterocyte-like cell line derived from a human colonic adenocarcinoma, as an intestinal model. Ecto-ALP activity and N-[methyl-3H]-4-phenylpyridinium acetate (3H-MPP+) uptake were evaluated in intact Caco-2 cells. T+ and TPP were able to increase ecto-ALP activity, with an equal potency, and to decrease 3H MPP+ apical uptake, with a similar potency. The effects of both compounds on ecto ALP activity and 3H-MPP+ uptake were concentration-dependent. The results suggest that the effect of T+ and TPP on ecto-ALP activity may lead to inhibition of the intestinal absorption of other organic cations present in the diet. Another important conclusion is that the intestinal absorption of T+ may occur through hEMT, in Caco-2 cells. PMID- 14527823 TI - Prophylaxis against lipopolysaccharide-induced liver injuries by lipoic acid in rats. AB - Lipopolysaccharide (LPS) is a major cell wall molecule of Gram-negative bacteria known to stimulate the synthesis and secretion of several metabolites, such as reactive oxygen species, from phagocytes that play an important role in the pathogenesis of tissue injuries. In this study, the prophylactic effect of the antioxidant lipoic acid was evaluated in an animal acute organ injury model. Animals were pre-treated intraperitoneally with lipoic acid (50 mg kg(-1) body weight) or saline; 3 h later, pretreated animals were challenged intravenously with LPS (Escherichia coli 0111:B4, 1.0 mg kg(-1) body weight) or saline and killed 21 h later. Saline-pretreated animals challenged with LPS were extensively damaged in the liver, as evidenced by an increase in plasma alanine and aspartate aminotransferase activities. Also, LPS injection to saline-pretreated animals resulted in significant increases in plasma tumour necrosis factor-alpha (TNFalpha) and nitric oxide (NO) concentrations, suggestive of activation of the proinflammatory response. The LPS challenge to saline-pretreated animals also increased hepatic myeloperoxidase activity as well as protease and chloramine levels, suggestive of neutrophil infiltration and activation of the inflammatory response. In addition, the involvement of oxidative stress was evident, because a significant increase in lipid peroxidation was observed in the livers of saline pretreated animals challenged with LPS. The administration of lipoic acid prior to LPS challenge resulted in a significant alleviation of liver injuries, evidenced by a general reversal of the altered biochemical indices toward normal among treated animals. These results indicate that lipoic acid may serve as a potentially effective prophylactic pharmacological agent in alleviating LPS induced tissue injuries. PMID- 14527824 TI - Effect of Praval bhasma (Coral calx), a natural source of rich calcium on bone mineralization in rats. AB - Praval bhasma (PB; Coral calx) is a natural source of rich calcium widely used in traditional system of Indian medicine as a supplement in the treatment of variety of bone metabolic disorders associated with calcium deficiency. The present study was designed to investigate the inhibitory effects of Praval bhasma on the progress of bone loss induced by ovariectomy and concurrent calcium deficiency (CD-OVX). Twenty-four female rats were ovariectomized, 12 sham operated, divided into three groups of 12 each, fed on low calcium diet (0.04% Ca) and treated either with vehicle or Praval bhasma (65 mg/kg body weight, twice a day) for 16 weeks. Compared to sham rats, CD-OVX animals showed an increase in urinary excretion of calcium (Ca) and phosphorus (P), decreased femoral weight and density which were significantly reversed in Praval bhasma treated animals. Measurement of cortical bone morphometric indices by CT-scanning technique showed increased medullary width and cross-sectional area, decreased periosteal area (PA), combined cortical thickness (CCT) and cortical area (CA)/periosteal area in CD-OVX animals compared to sham and PB-treated group. Scanning electron microscopy (SEM) study revealed porous and erosive appearance of the distal femur at the epiphysis and reduced Ca:P ratio in CD-OVX animals compared to sham and PB treated group. Ash weight, percent ash, ash Ca and ash P levels were lower in CD OVX animals than in sham or PB group. Histological examination of decalcified femurs showed narrowed and disappearance of trabeculae and widened medullary spaces and decreased impact strength as measured by impact test in CD-OVX animals compared to sham and PB-treated group. The present study concludes that Praval bhasma is effective in the prevention of calcium and estrogen deficient bone loss and justify the continuing use of this ayurvedic preparation in traditional system of Indian medicine for management of bone metabolic disorders such as osteoporosis. PMID- 14527825 TI - Comparative study of flavonoids in experimental models of inflammation. AB - The anti-inflammatory activities of flavonols (quercetin, rutin and morin) and flavanones (hesperetin and hesperidin) were investigated in animal models of acute and chronic inflammation. Rutin was only effective in the chronic process, principally in adjuvant arthritis. On neurogenic inflammation induced by xylene, only the flavanones were effective; besides, these compounds were the most effective on subchronic process. The most important compound in reducing paw oedema induced by carrageenan was quercetin. PMID- 14527826 TI - Changes in norepinephrine and epinephrine concentrations in adrenal gland of the rats submitted to acute immobilization stress. AB - Catecholamines act as neurotransmitters and hormones. Studies conducted to understand the synthesis and metabolism of these monoamines during stress have been the main concern of many authors. This work proposes to investigate the time course of changes in epinephrine and norepinephrine concentration in adrenal gland obtained from rats submitted to acute immobilization stress. The results of the present study indicate that acute immobilization stress during 5 and 15 min did not provoke changes in epinephrine and norepinephrine concentrations in adrenal gland in relation to the control group. Such results are justified due to the short time of the stress, showing that the stress did not provoke physiological alteration. The epinephrine and norepinephrine concentrations in adrenal gland increased significantly after the immobilization session in stressed groups during 30 and 50 min as compared to control group. This increase probably is due to the emotional component of the immobilization stress. In this way, we suggested that the immobilization stress provoke increase in the biosynthesis of catecholamines in the adrenal gland from rats. However, the results shows that a maximum increase is reached at 30 min of immobilization stress and then a decrement of catecholamines levels starts at 50 min of the experimental design. This decline in catecholamines level may be consequence of adaptation to stress situations, an increase of the activity of the uptake systems and/or metabolization of catecholamines. In conclusion, these results suggest an effective participation of the adrenal glands to maintain the homeostasis of organism to the stressful conditions. PMID- 14527828 TI - Relationship between activation of microsomal glutathione S-transferase and metabolism behavior of chlorambucil. AB - Chlorambucil (4-[p-[bis [2-chloroethyl] amino] phenyl]-butanoic acid; CMB) is a bifunctional alkylating agent that exhibits acquired drug resistance upon repeated dosing in human. This compound reacts with glutathione (gamma glutamylcysteinylglycine, GSH) both non-enzymatically and enzymatically in the presence of glutathione S-transferases (GSTs) to produce several GSH conjugates. The formation of GSH conjugates of this class of alkylating agents may play a role in the development of acquired drug resistance. In this study, the effect of CMB on microsomal GST (mGST) activity and the catalytic effect of mGST on the formation of GSH conjugates of CMB were investigated. mGST was activated by CMB in a concentration- and time-dependent manner. NEM failed to reactivate the CMB pretreated mGST. This suggests that mGST could be activated by CMB, possibly via reacting with the single cysteine (Cys49) in mGST. The resulting conjugates of GSH and CMB were characterized by a combination of lipid chromatography/electrospray ionization mass spectrometry (LC/ESI/MS). mGST presented a strong catalytic effect on the reaction as determined by the increase of the diglutathionyl derivative, 4-[p-[bis[2-S-glutathionylethyl] amino]phenyl] butanoic acid (CMBSG2), monoglutathionyl derivatives, 4-[p-[N-2-chloroethyl, N-2 S-glutathionylethyl] amino]phenyl]-butanoic acid (CMBSG) and 4-[p-[N-2-S glutathionylethyl, N-2-hydroxyethyl] amino]phenyl]-butanoic acid (CMBSGOH) and the decrease of CMB. mGST which was stimulated approximately 2.5-fold with CMB had a stronger catalytic effect. These results suggest the potential role of the activation of mGST in the CMB inactivation and in the development of acquired drug resistance. PMID- 14527827 TI - Pharmacological actions of a novel and selective dopamine D3 receptor antagonist, KCH-1110. AB - 1-(2-ethoxy-phenyl)-4-[3-(3-thiophen-2-yl-isoxazolin-5-yl)-propyl]-piperazine (KCH-1110), has a high affinity for human dopamine D3 (hD3) receptor (Ki=1.28 nM) with about 90-fold selectivity over the human dopamine D2L (hD2L) receptor. Antipsychotic or antidopaminergic activity of KCH-1110 was investigated in the models for the positive symptoms of schizophrenia, apomorphine-induced climbing and cocaine-induced hyperlocomotion, in mice. Intraperitoneal (i.p.) or oral (p.o.) administration of KCH-1110 potently inhibited the apomorphine-induced cage climbing without any rotarod ataxia in mice. Cocaine-induced hyperactivity was also antagonised by KCH-1110. In addition, KCH-1110 attenuated the hypothermia induced by a selective dopamine D3 agonist, 7-OH-DPAT in mice. KCH-1110 did not induce catalepsy in mice, but at much higher doses only a slight catalepsy response was shown. Although high doses of KCH-1110 significantly enhanced serum prolactin secretion in rats, low dose of KCH-1110 did not increase prolactin levels in rats. The present studies, therefore, suggest that KCH-1110 is a potent and relatively selective dopamine D3 receptor antagonist with antipsychotic actions. PMID- 14527829 TI - Protective effect of arabic gum against acetaminophen-induced hepatotoxicity in mice. AB - Overdose of acetaminophen, a widely used analgesic drug, can result in severe hepatotoxicity and is often fatal. This study was undertaken to examine the effects of arabic gum (AG), which is commonly used in processed foods, on acetaminophen-induced hepatotoxicity in mice. Mice were given arabic gum orally (100 g l(-1)) 5 days before a hepatotoxic dose of acetaminophen (500 mg kg(-1)) intraperitoneally. Arabic gum administration dramatically reduced acetaminophen induced hepatotoxicity as evidenced by reduced serum alanine (ALT) and aspartate aminotransferase (AST) activities. Acetaminophen-induced hepatic lipid peroxidation was reduced significantly by arabic gum pretreatment. The protection offered by arabic gum does not appear to be caused by a decrease in the formation of toxic acetaminophen metabolites, which consumes glutathione, because arabic gum did not alter acetaminophen-induced hepatic glutathione depletion. Acetaminophen increased nitric oxide synthesis as measured by serum nitrate plus nitrite at 4 and 6 h after administration and arabic gum pretreatment significantly reduced their formation. In conclusion, arabic gum is effective in protecting mice against acetaminophen-induced hepatotoxicity. This protection may involve the reduction of oxidative stress. PMID- 14527830 TI - The effect of manganese chloride on gentamicin-induced nephrotoxicity in rats. AB - The aim of this study was to investigate the effects of manganese chloride on gentamicin-induced nephrotoxicity in rats. Thirty-six adult Wistar Albino rats were divided into six equal groups. They were injected with gentamicin sulfate (100 mg kg(-1) per day i.p.) and manganese chloride (2 or 20 mg kg(-1) per day i.p.) and gentamicin together with manganese chloride for 6 days. The animals were killed 24 h after the last injection. Nephrotoxicity was biochemically and histopathologically evaluated. The concentrations of creatinine, urea, sodium and potassium in plasma, malondialdehyde (MDA) and reduced glutathione (GSH) levels, glutathione peroxidase (GSH-Px) and catalase (CAT) activities in kidney tissue were determined. Administration of gentamicin to rats induced a marked renal failure, characterized with a significant increase in plasma creatinine and urea concentrations. A significant increase in kidney MDA and a decrease in GSH concentrations were observed in gentamicin-treated rats. No change was observed in the activities of GSH-Px and CAT in rats treated with gentamicin alone. Administration of the low dose of manganese (Mn2+) produced amelioration in biochemical indices of nephrotoxicity in plasma and kidney tissue when compared to gentamicin group. The histological signs of renal proximal tubules followed a similar pattern. The high dose of Mn2+ (20 mg kg(-1)) caused an opposite effect on nephrotoxicity induced by gentamicin, causing exacerbation in the tubular necrosis. The results suggest that low dose of Mn2+ may have an antioxidant effect in kidneys of gentamicin administrated rats, but its high doses had no beneficial effect. PMID- 14527831 TI - Bioavailability of granulocyte colony-stimulating factor administered enterally to suckling mice. AB - The developing fetal and neonatal gastrointestinal (GI) tract is influenced by many growth factors, including epidermal growth factor (EGF), insulin-like growth factor (IGF), transforming growth factor (TGF), and erythropoietin (Epo). Granulocyte colony-stimulating factor (G-CSF), typically regarded as a hematopoietic growth factor, might also be included because it exists in high concentrations in amniotic fluid, colostrum, and human milk, and because granulocyte CSF receptors (G-CSF-R) are abundantly expressed on the villous enterocytes of the developing intestine. As a first step toward understanding whether the effects of G-CSF on the GI tract were local or systemic, we sought to determine whether recombinant human G-CSF (rhG-CSF) administered enterally to suckling mice, is absorbed into the circulation, and if so, whether the G-CSF-R is essential for this absorption. We enterally administered rhG-CSF to suckling mice, selecting a daily dose based on the amount of G-CSF normally swallowed by the fetus and neonate (3 ng), or in other mice, a dose of G-CSF 100 times larger (300 ng). Pups were tested at either 5-7 days of age, or at 14-16 days of age. C57BL/6 x 129SvJ mice were used. Some mice had a targeted null mutation in the G CSF-R gene, producing a non-functional G-CSF-R protein. At intervals following the enteral G-CSF dosing, G-CSF concentrations in plasma were measured by specific ELISA. The bioavailability of G-CSF was invariably <1%, regardless of the dose of rhG-CSF given, the age of the pups, or whether they had a functional G-CSF-R. After enteral administration of rhG-CSF to suckling mice, only minimal quantities of G-CSF are absorbed into the circulation, and the G-CSF-R is not essential for this absorption. PMID- 14527832 TI - Lithium dose prediction based on 24 hours single dose levels: a prospective evaluation. AB - The authors present the results of the utilization of a pharmacokinetic prediction test for lithium posology. Based on a single point (plasma lithium determination 24 h after a single dose) such a test aims to adapt the posology as soon as the second day of treatment rather than after one week as clinicians must wait for a steady state to be achieved. Built on the previous work of Perry, the test targeted the plasma lithium level at 0.8 +/- 0.1 mmol l(-1). Thirty-one patients took part in the study. There were two drop-out cases and the results were available for 29 patients: among them, 51% had their plasma level in the targeted zone. Although there was no control group, the prediction test often allowed us to use a higher dose than the usual fixed dose whose amount is limited by the risk of overdosing for the slower metabolizers. PMID- 14527833 TI - Plasma pooling methodology as a faster and cheaper tool to evaluate bioequivalence of rifampicin component of FDCs of antitubercular drugs. AB - Rifampicin is one of the most important first line drugs used in fixed dose combinations (FDCs) of antitubercular drugs. The chances of reduced bioavailability of rifampicin from FDCs necessitated its evaluation against standard formulations of individual drugs in bioequivalence studies. This study was undertaken to evaluate the importance of plasma pooling methodology as a rapid and cheaper tool to evaluate bioequivalence of rifampicin component of FDCs of antitubercular drugs. Plasma samples of volunteers obtained from bioequivalence studies were pooled according to volunteer and time-wise pooling. Area under the plasma concentration versus time curves (AUCs) of rifampicin was reduced in the pooled plasma samples when compared to the individual samples. However, the ratio of AUCs of FDCs to that of separate formulations was found to be comparable for both the individual samples and pooled samples data. Concentration-time profiles of rifampicin obtained after time-wise pooling were subjected to non-compartmental analysis for determination of pharmacokinetic parameters. Whereas bioequivalence estimates were determined using individual AUC values obtained from volunteer-wise pooling. Results indicated the possibility of using plasma pooling methodology in bioequivalence estimation of rifampicin to simplify and accelerate the registration process. PMID- 14527834 TI - Embracing the local: enriching scientific research, education, and outreach on the Texas-Mexico border through a participatory action research partnership. AB - Cameron Park, Texas, is a colonia (an isolated, unincorporated rural settlement without municipal improvements) on the Texas-Mexico border in the Lower Rio Grande Valley, in Cameron County near Brownsville, Texas. Cameron Park has a population of 5,961 residents, 99.3% of whom are Hispanic. The annual median income is 16,934 US dollars, about one-half of the state median. Fifty-eight percent of families generally and 68% of those with children younger than 5 years have incomes below poverty level. Cameron Park resides geographically in a region where agriculture has been, and continues to be, a dominant industry, a fact consistent with the intensive use of pesticides and increased potential for air, water, and ground contamination. The practice of good environmental health is extremely difficult under these conditions. In 1999 the Texas A&M University Center for Housing and Urban Development's Colonias Program and the Center for Environmental and Rural Health teamed up to create an environmental health education and outreach program called the Cameron Park Project (CPP). The CPP focused on how to reduce potential environmental exposures associated with human illness by providing residents with scientifically sound information on positive health practices and how to deal with environmental hazards. In this article we discuss the research methodology used in the CPP, a methodology specifically chosen to address four challenges presented by colonias to conducting valid and reliable research. PMID- 14527835 TI - Risk management and precaution: insights on the cautious use of evidence. AB - Risk management, done well, should be inherently precautionary. Adopting an appropriate degree of precaution with respect to feared health and environmental hazards is fundamental to risk management. The real problem is in deciding how precautionary to be in the face of inevitable uncertainties, demanding that we understand the equally inevitable false positives and false negatives from screening evidence. We consider a framework for detection and judgment of evidence of well-characterized hazards, using the concepts of sensitivity, specificity, positive predictive value, and negative predictive value that are well established for medical diagnosis. Our confidence in predicting the likelihood of a true danger inevitably will be poor for rare hazards because of the predominance of false positives; failing to detect a true danger is less likely because false negatives must be rarer than the danger itself. Because most controversial environmental hazards arise infrequently, this truth poses a dilemma for risk management. PMID- 14527836 TI - Historical pesticide exposure in California using pesticide use reports and land use surveys: an assessment of misclassification error and bias. AB - We used California's Pesticide Use Report (PUR) and land-use survey data to conduct a simulation study evaluating the potential consequences of misclassifying residential exposure from proximity to agricultural pesticide application in health effect studies. We developed a geographic model linking the PUR with crop location data from land-use surveys to assess the impact of exposure misclassification from simpler exposure models based solely on PUR or land-use data. We simulated the random selection of population controls recruited into a hypothetical case-control study within an agricultural region. Using residential parcel data, we derived annual exposure prevalences, sensitivity, and specificity for five pesticides and relied on the PUR plus land-use model as the "gold standard." Based on these estimates, we calculated the attenuation of prespecified true odds ratios (ORs), assuming nondifferential exposure misclassification. True ORs were severely attenuated a) when residential exposure status was based on a larger geographic area yielding higher sensitivity but low specificity for exposure, in contrast to relying on a smaller area and increasing specificity; b) for less frequently applied pesticides; and c) with increasing mobility of residents among the study population. Considerable effect estimate attenuation also occurred when we used residential distance to crops as a proxy for pesticide exposure. Finally, exposure classifications based on annual instead of seasonal summaries of PUR resulted in highly attenuated ORs, especially during seasons when applications of specific pesticides were unlikely to occur. These results underscore the importance of increasing the spatiotemporal resolution of pesticide exposure models to minimize misclassification. PMID- 14527837 TI - Antibiotics in dust originating from a pig-fattening farm: a new source of health hazard for farmers? AB - Pig-house dust originates from feed, bedding, feces, and the animals themselves. If the animals receive drugs such as antibiotics, residues of these substances may occur in manure, in the air, or on surfaces of the respective animal house. In a retrospective study, we investigated dust samples collected during two decades from the same piggery for the occurrence of various antibiotics. In 90% of these samples, we detected up to five different antibiotics, including tylosin, various tetracyclines, sulfamethazine, and chloramphenicol, in total amounts up to 12.5 mg/kg dust. High dust exposure in animal confinement buildings is believed to be a respiratory health hazard because of the high content of microorganisms, endotoxins, and allergens. Further risks may arise from the inhalation of dust contaminated with a cocktail of antibiotics. Apart from that, our data provide first evidence for a new route of entry for veterinary drugs in the environment. PMID- 14527838 TI - Measurement of brevetoxin levels by radioimmunoassay of blood collection cards after acute, long-term, and low-dose exposure in mice. AB - We developed a radioimmunoassay (RIA) using a sheep anti-brevetoxin antiserum to evaluate detection of brevetoxin on blood collection cards from mice treated with the brevetoxin congener PbTx-3. The RIA has high affinity for PbTx-3 [half maximal effective concentration (EC(50)) +/- SE = 1.2 +/- 0.2 nM; n = 10] and recognizes both type 1 and type 2 brevetoxins, but not ciguatoxin. Direct comparison of the RIA with a radiolabeled [(3)H]-PbTx-3 receptor-binding assay (RBA) revealed excellent sensitivity, congener selectivity, and minimal interference from blood matrix. We first analyzed blood samples from an acute time course exposure, using a maximal nonlethal dose [180 microg/kg body weight (bw)] for 0.5, 1, 2, 4, and 24 hr. Mean blood brevetoxin levels were 36 nM at 30 min and stayed above 20 nM during the 1-4 hr time points. We next analyzed blood brevetoxin levels after longer exposure (0.5, 1, 2, 3, 4, or 7 days). Mean blood brevetoxin levels were 26.0 nM at 0.5 days, decreased to 8.2 nM at 1.0 day, and maintained a significant level (p < 0.05) of 1.3 nM at day 2. We next determined the lowest measurable dose using increasing concentrations of PbTx-3 (10-300 micro g/kg bw). Analysis of the blood samples at 60 min revealed a linear relationship between administered and internal doses (r(2) = 0.993). All doses of brevetoxin administered were detectable at 1 hr, with significant levels found for the lowest administered dose of 10 micro g/kg bw--a dose that was 10-fold lower than the lowest observable effect level. This RIA provides an optimal first tier detection of brevetoxin from blood collection cards and, used in combination with the RBA and liquid chromatography-mass spectrometry, should provide a complete panel of methods to biomonitor brevetoxin exposure. PMID- 14527839 TI - Age- and sex-dependent distribution of persistent organochlorine pollutants in urban foxes. AB - The colonization of urban and suburban habitats by red foxes (Vulpes vulpes) provides a novel sentinel species to monitor the spread of anthropogenic pollutants in densely populated human settlements. Here, red foxes were collected in the municipal territory of Zurich, Switzerland, and their perirenal adipose tissue was examined for persistent organochlorine residues. This pilot study revealed an unexpected pattern of contamination by polychlorinated biphenyls (PCBs), with significantly higher levels of the predominant congeners PCB-138, PCB-153, and PCB-180 in juvenile foxes relative to adult animals. Further data analysis demonstrated that the observed difference was attributable to an age dependent reduction of PCB concentrations in females, whereas male foxes retained approximately the same PCB burden throughout their life span. A similar sex related bias between population members has been observed, primarily in marine mammals. Interestingly, the reduction of organochlorine contents with progressive age is reminiscent of human studies, where an extensive maternal transfer of xenobiotics to the offspring has been shown to result in increased exposure levels of infants relative to adults. To our knowledge, this is the first example of an urban wildlife species that faithfully reflects the dynamic distribution of toxic contaminants in the corresponding human population. Suburban and urban foxes occupy habitats in close proximity to humans, depend on anthropogenic food supplies, are relatively long-lived and readily available for sampling, can be easily aged and sexed, have a limited home range, and, therefore, meet several important requirements to serve as a surrogate species for the assessment of toxic health hazards. PMID- 14527840 TI - Associations of renal function with polymorphisms in the delta-aminolevulinic acid dehydratase, vitamin D receptor, and nitric oxide synthase genes in Korean lead workers. AB - We analyzed data from 798 lead workers to determine whether polymorphisms in the genes encoding delta-aminolevulinic acid dehydratase (ALAD), endothelial nitric oxide synthase (eNOS), and the vitamin D receptor (VDR) were associated with or modified relations of lead exposure and dose measures with renal outcomes. Lead exposure was assessed with job duration, blood lead, dimercaptosuccinic acid (DMSA)-chelatable lead, and tibia lead. Renal function was assessed with blood urea nitrogen (BUN), serum creatinine, measured creatinine clearance, calculated creatinine clearance and urinary N-acetyl-beta-D-glucosaminidase (NAG), and retinol-binding protein. Mean (+/- SD) tibia lead, blood lead, and DMSA chelatable lead levels were 37.2 +/- 40.4 microg/g bone mineral, 32.0 +/- 15.0 microg/dL, and 767.8 +/- 862.1 microg/g creatinine, respectively. After adjustment, participants with the ALAD(2) allele had lower mean serum creatinine and higher calculated creatinine clearance. We observed effect modification by ALAD on associations between blood lead and/or DMSA-chelatable lead and three renal outcomes. Among those with the ALAD(1-2) genotype, higher lead measures were associated with lower BUN and serum creatinine and higher calculated creatinine clearance. Participants with the eNOS variant allele were found to have higher measured creatinine clearance and BUN. In participants with the Asp allele, longer duration working with lead was associated with higher serum creatinine and lower calculated creatinine clearance and NAG; all were significantly different from relations in those with the Glu/Glu genotype except NAG (p = 0.08). No significant differences were seen in renal outcomes by VDR genotype, nor was consistent effect modification observed. The ALAD findings could be explained by lead-induced hyperfiltration. PMID- 14527841 TI - Hearing loss in workers exposed to carbon disulfide and noise. AB - Simultaneous exposure to carbon disulfide and noise may have a combined effect on hearing impairment. In this study we investigated hearing loss in 131 men with exposure to noise [80-91 A-weighted decibels; dB(A)] and CS(2) (1.6-20.1 ppm) in a viscose rayon plant. These men were compared with 105 men in the adhesive tape and electronic industries who were exposed to noise only and with 110 men employed in the administrative office of the rayon plant who were exposed to low noise and no CS(2). We conducted interviews to obtain sociodemographic information and exposure assessments, and we performed physical examinations, including hearing tests. Results showed that the prevalence of hearing loss of > 25 dB hearing loss (dBHL) in rayon workers (67.9%) was much higher than that in administrative workers (23.6%) and in the adhesive tape and electronic industrial workers (32.4%). Hearing loss occurred mainly for speech frequencies of 0.5, 1, and 2 kHz. When the CS(2) exposure was measured by the product of CS(2 )exposure level and employment years, the adjusted odds ratios of hearing loss of > 25 dBHL in rayon workers, compared with administrative workers, were 3.8 [95% confidence interval (CI), 1.5-9.4] for those with the exposure of 37-214 year-ppm, 14.2 (95% CI, 4.4-45.9) with 215-453 year-ppm exposure, and 70.3 (95% CI, 8.7-569.7) with exposure of > 453 year-ppm. The study suggests that CS(2) exposure enhances human hearing loss in a noisy environment and mainly affects hearing in lower frequencies. PMID- 14527843 TI - Comparable measures of cognitive function in human infants and laboratory animals to identify environmental health risks to children. AB - The importance of including neurodevelopmental end points in environmental studies is clear. A validated measure of cognitive function in human infants that also has a homologous or parallel test in laboratory animal studies will provide a valuable approach for large-scale studies. Such a comparable test will allow researchers to observe the effect of environmental neurotoxicants in animals and relate those findings to humans. In this article, we present the results of a review of post-1990, peer-reviewed literature and current research examining measures of cognitive function that can be applied to both human infants (0-12 months old) and laboratory animals. We begin with a discussion of the definition of cognitive function and important considerations in cross-species research. We then describe identified comparable measures, providing a description of the test in human infants and animal subjects. Available information on test reliability, validity, and population norms, as well as test limitations and constraints, is also presented. PMID- 14527842 TI - Measurement of offline exhaled nitric oxide in a study of community exposure to air pollution. AB - As part of a large panel study in Seattle, Washington, we measured levels of exhaled nitric oxide (eNO) in children's homes and fixed-site particulate matter with aerodynamic diameters of 2.5 micro m or less (PM(2.5)) outside and inside the homes as well as personal PM(2.5) during winter and spring sessions of 2000 2001. Nineteen subjects 6-13 years of age participated; 9 of the 19 were on inhaled corticosteroid (ICS) therapy. Exhaled breath measurements were collected offline into a Mylar balloon for up to 10 consecutive days. Mean eNO values were 19.1 (SD +/- 11.4) ppb in winter sessions and 12.5 +/- 6.6 ppb in spring sessions. Fixed-site PM(2.5) mean concentrations were 10.1 +/- 5.7 microg/m(3) outside homes and 13.3 +/- 1.4 inside homes; the personal PM(2.5) mean was 13.4 +/- 3.2 microg/m(3). We used a linear mixed-effects model with random intercept and an interaction term for medications to test for within-subject-within-session associations between eNO and various PM(2.5) values. We found a 10 microg/m(3) increase in PM(2.5) from the outdoor, indoor, personal, and central-site measurements that was associated with increases in eNO in all subjects at lag day zero. The effect was 4.3 ppb [95% confidence interval (CI), 1.4-7.29] with the outdoor monitor, 4.2 ppb (95% CI, 1.02-7.4) for the indoor monitor, 4.5 ppb (95% CI, 1.02-7.9) with the personal monitor, and 3.8 ppb (95% CI, 1.2-6.4) for the central monitors. The interaction term for medication category (ICS users vs. nonusers) was significant in all analyses. These findings suggest that eNO can be used as an assessment tool in epidemiologic studies of health effects of air pollution. PMID- 14527844 TI - Cumulative organophosphate pesticide exposure and risk assessment among pregnant women living in an agricultural community: a case study from the CHAMACOS cohort. AB - Approximately 230,000 kg of organophosphate (OP) pesticides are applied annually in California's Salinas Valley. These activities have raised concerns about exposures to area residents. We collected three spot urine samples from pregnant women (between 1999 and 2001) enrolled in CHAMACOS (Center for the Health Assessment of Mothers and Children of Salinas), a longitudinal birth cohort study, and analyzed them for six dialkyl phosphate metabolites. We used urine from 446 pregnant women to estimate OP pesticide doses with two deterministic steady-state modeling methods: method 1, which assumed the metabolites were attributable entirely to a single diethyl or dimethyl OP pesticide; and method 2, which adapted U.S. Environmental Protection Agency (U.S. EPA) draft guidelines for cumulative risk assessment to estimate dose from a mixture of OP pesticides that share a common mechanism of toxicity. We used pesticide use reporting data for the Salinas Valley to approximate the mixture to which the women were exposed. Based on average OP pesticide dose estimates that assumed exposure to a single OP pesticide (method 1), between 0% and 36.1% of study participants' doses failed to attain a margin of exposure (MOE) of 100 relative to the U.S. EPA oral benchmark dose(10) (BMD(10)), depending on the assumption made about the parent compound. These BMD(10) values are doses expected to produce a 10% reduction in brain cholinesterase activity compared with background response in rats. Given the participants' average cumulative OP pesticide dose estimates (method 2) and regardless of the index chemical selected, we found that 14.8% of the doses failed to attain an MOE of 100 relative to the BMD(10) of the selected index. An uncertainty analysis of the pesticide mixture parameter, which is extrapolated from pesticide application data for the study area and not directly quantified for each individual, suggests that this point estimate could range from 1 to 34%. In future analyses, we will use pesticide-specific urinary metabolites, when available, to evaluate cumulative OP pesticide exposures. PMID- 14527845 TI - Integrated pest management in an urban community: a successful partnership for prevention. AB - Pesticides, applied in large quantities in urban communities to control cockroaches, pose potential threats to health, especially to children, who have proportionately greater exposures and unique, developmentally determined vulnerabilities. Integrated pest management (IPM) relies on nonchemical tools- cleaning of food residues, removal of potential nutrients, and sealing cracks and crevices. Least toxic pesticides are used sparingly. To evaluate IPM's effectiveness, the Mount Sinai Children's Environmental Health and Disease Prevention Research Center, in partnership with two community health centers in East Harlem, New York City (NY, USA), undertook a prospective intervention trial. Families (n = 131) enrolled when mothers came to the centers for prenatal care. Household cockroach infestation was measured by glue traps at baseline and 6 months afterward. The intervention group received individually tailored IPM education, repairs, least-toxic pest control application, and supplies, with biweekly pest monitoring for 2 months and monthly for 4 months. The control group, residing in East Harlem and demographically and socioeconomically similar to the intervention group, received an injury prevention intervention. The proportion of intervention households with cockroaches declined significantly after 6 months (from 80.5 to 39.0%). Control group levels were essentially unchanged (from 78.1 to 81.3%). The cost, including repairs, of individually tailored IPM was equal to or lower than traditional chemically based pest control. These findings demonstrate that individually tailored IPM can be successful and cost-effective in an urban community. PMID- 14527846 TI - Acute occupational disinfectant-related illness among youth, 1993-1998. AB - Working youths face many safety and health risks. Among these risks are those posed by disinfectant exposures. In this study we describe acute occupational disinfectant-related illness among youth. Data on U.S. children younger than 18 years with acute occupational disinfectant-related illnesses between 1993 and 1998 were collected from the Toxic Exposure Surveillance System and from the California Department of Pesticide Regulation. We analyzed data from persons with exposures who met the case definition for acute occupational disinfectant-related illness. The case definition required onset of new adverse health effects that were both temporally related to a disinfectant exposure and consistent with the known toxicology of the disinfectant. We calculated incidence rates of acute occupational disinfectant-related illness among youths 15-17 years old and incidence rate ratios to compare these rates with those of adults 25-44 years old. We found 307 children with disinfectant-related illnesses. The average annual incidence rate was 16.8/billion hours worked with a relative risk compared with adults of 4.14 (95% confidence interval, 3.66-4.68). Most illnesses were of mild severity (78%). There were no fatalities. Hypochlorites (e.g., bleach) were responsible for 45% of the illnesses. Among the 206 cases where the responsible disinfectant's U.S. Environmental Protection Agency toxicity category was known, 80% were in category I (highest toxicity level). These findings suggest the need for greater efforts to prevent adolescent acute occupational disinfectant-related illness. This may require strengthening regulations and enforcement as well as increased educational efforts directed at employers, youths, parents, school officials, and physicians. Better mechanisms for reporting and tracking chemical illnesses among working adolescents are also needed. PMID- 14527847 TI - Time trends of persistent organic pollutants and heavy metals in umbilical cord blood of Inuit infants born in Nunavik (Quebec, Canada) between 1994 and 2001. AB - Inuit inhabitants of Nunavik (northern Quebec, Canada) consume great quantities of marine food and are therefore exposed to high doses of food chain contaminants. In this study, we report the time trends of persistent organic pollutants, mercury, and lead in umbilical cord blood of infants from three communities of the east coast of Hudson Bay in Nunavik. We analyzed 251 cord blood samples collected from 1994 through 2001 for polychlorinated biphenyls (PCBs), dichlorodiphenyl trichloroethane (DDT), dichlorodiphenyl dichloroethylene (DDE), hexachlorobenzene (HCB), chlordanes, lead, and mercury. Using an exponential model, we found strongly significant decreasing trends for PCBs (7.9% per year, p < 0.001), DDE (9.1% per year, p < 0.001), DDT (8.2% per year, p < 0.001), and HCB (6.6% per year, p < 0.01). No significant trends were detected for chlordanes. A significant reduction of lead and mercury concentrations was found, but there was no clear linear or exponential trend. The decreases observed could be explained by a decrease in food contamination, by changes in dietary habits, or, most likely, by a combination of both. PMID- 14527848 TI - Vitamin D receptor Fok1 polymorphism and blood lead concentration in children. AB - Variation in blood lead concentration is caused by a complex interaction of environmental, social, nutritional, and genetic factors. We evaluated the association between blood lead concentration and a vitamin D receptor (VDR) gene polymorphism. Environmental samples and blood were analyzed for lead, nutritional and behavioral factors were assessed, and VDR -Fok1 genotype was determined in 245 children. We found a significant interaction between floor dust lead and genotype on blood lead concentration. For every 1 microg/ft(2) increase in floor dust, children with VDR -FF genotype had a 1.1% increase in blood lead [95% confidence interval (CI), 0.69-1.5], VDR -Ff, 0.53% increase (95% CI, 0.1-0.92), and VDR -ff, 3.8% increase (95% CI, 1.2-6.3); however, at floor dust levels < 10 microg/ft(2), children with VDR -ff had the lowest blood lead concentrations. These data suggest that VDR -Fok1 is an effect modifier of the relationship of floor dust lead exposure and blood lead concentration. PMID- 14527849 TI - Prenatal PCB exposure, the corpus callosum, and response inhibition. AB - The present study reports the association between prenatal exposure to polychlorinated biphenyls (PCBs), the corpus callosum, and response inhibition in children who are 4.5 years old. Children (n = 189) enrolled in the Oswego study were tested using a continuous performance test. We measured (square millimeters) the splenium of the corpus callosum, a pathway implicated in the regulation of response inhibition, using magnetic resonance imaging. Results indicated a dose dependent association between cord blood PCBs and errors of commission. Splenium size but not other brain areas predicted errors of commission (r(2) = 0.20), with smaller size associated with more errors of commission. There was an interaction between splenium size and PCB exposure. The smaller the splenium, the larger the association between PCBs and errors of commission. If the association between PCBs and response inhibition is indeed causal, then children with suboptimal development of the splenium are particularly vulnerable to these effects. These data await replication. PMID- 14527850 TI - Assessing the effects of endocrine disruptors in the National Children's Study. AB - Children are uniquely vulnerable to toxic chemicals in the environment. Among the environmental toxicants to which children are at risk of exposure are endocrine disruptors (EDs)--chemicals that have the capacity to interfere with hormonal signaling systems. EDs may alter feedback loops in the brain, pituitary, gonads, thyroid, and other components of the endocrine system. They can affect development. Effects of EDs have been described in wildlife populations, in animals exposed experimentally, and to a more limited extent in humans. Mechanisms of action of EDs are increasingly being elucidated, and genetic polymorphisms that convey differential susceptibility to EDs are beginning to be explored. It is hypothesized that in utero and early childhood exposures to EDs may be responsible, at least in part, for decreases in semen quality; increasing incidence of congenital malformations of the reproductive organs, such as hypospadias; increasing incidence of testicular cancer; and acceleration of onset of puberty in females. The National Children's Study (NCS) will provide a unique opportunity to test the validity of these hypotheses in the context of a large prospective multi-year epidemiologic investigation. It will be essential in the NCS to assess exposures to a range of putative natural and synthetic EDs, to assess outcomes possibly due to ED exposure, to examine the potential interplay between EDs and genetic polymorphisms, and to seek links between ED exposures in early life and endocrine, reproductive, neurobehavioral, and other outcomes throughout the life span. PMID- 14527851 TI - Exposure assessment for endocrine disruptors: some considerations in the design of studies. AB - In studies designed to evaluate exposure-response relationships in children's development from conception through puberty, multiple factors that affect the generation of meaningful exposure metrics must be considered. These factors include multiple routes of exposure; the timing, frequency, and duration of exposure; need for qualitative and quantitative data; sample collection and storage protocols; and the selection and documentation of analytic methods. The methods for exposure data collection and analysis must be sufficiently robust to accommodate the a priori hypotheses to be tested, as well as hypotheses generated from the data. A number of issues that must be considered in study design are summarized here. PMID- 14527853 TI - Obesity in the 21st century. PMID- 14527854 TI - Correspondence: preformed biomarkers in produce inflate human organophosphate exposure assessments. PMID- 14527856 TI - Pesticide exposures and children's risk tradeoffs. PMID- 14527852 TI - An approach to assessment of endocrine disruption in the National Children's Study. AB - In this article we consider the importance of assessing endocrine disruption in a large new cohort that has been proposed, the National Children's Study (NCS). We briefly review evidence that endocrine disruption is a potentially important hypothesis for human studies and weigh the need to assess endocrine disruption in the NCS. We note the salient features of earlier, similar cohort studies that serve as reference points for the design of the NCS. Finally, we discuss features of the NCS that would allow or enhance assessment of endocrine disruption, even if endocrine disruption were not a primary hypothesis motivating the study. At this time, the evidence supporting endocrine disruption in humans with background level exposures is not strong. Thus, a compelling rationale for the NCS will probably need to be based on core hypotheses that focus on other issues. Nonetheless, if properly designed, the NCS could serve as an excellent resource for investigating future hypotheses regarding endocrine disruption. PMID- 14527857 TI - Corn and corn-derived products: sources of endocrine disruptors. PMID- 14527860 TI - Obesity: a weighty issue for children. PMID- 14527861 TI - Adjusting for youth: updated cancer risk guidelines. PMID- 14527862 TI - Dedicated outdoor air systems: rx for sick buildings. PMID- 14527864 TI - Matching health messages to information-processing styles: need for cognition and mammography utilization. AB - The use of tailored health communications has become a favored technique for persuading individuals to engage in health behaviors, such as screening mammography. This experiment examined the impact of tailoring persuasive health communications to one aspect of individuals' information-processing styles, that of the need for cognition (NFC), the enjoyment of thinking deeply about issues. To determine whether messages matched to an individual's NFC are more influential than mismatched messages, 602 women who called the Cancer Information Service (CIS) of the National Cancer Institute were asked to participate in an experiment at the end of their service call. They were assigned randomly to receive 1 of 2 phone messages promoting mammography use and a similarly tailored pamphlet 1 month later. Messages matched to an individual's NFC were better at motivating mammography 6 months later among high-NFC women. After controlling for prior mammography utilization, age, worry, intentions, perceived norms, suggestions to get a mammogram, and marital status, the interaction between participant NFC and message type also approached statistical significance. The differential influence of these brief, tailored communications diminished after 12 months, however. PMID- 14527865 TI - Correspondence among patients' self-reports, chart records, and audio/videotapes of medical visits. AB - This pilot study examined the covariation of patients' self-reports of instrumental and affective aspects of communication during physician-patient visits with 2 other sources of data: medical chart records and audio/videotapes. Participants were 17 community-based (nonuniversity) primary-care physicians and 77 of their patients, ages 50 to 80. Patients were interviewed by telephone within 1 week after their medical visits. Thirty-five of these visits were audio- and videotaped. Patients were asked to report on their receipt of specific cancer screening in the previous 2 years, the occurrence of instrumental communication events during the visit (e.g., recommendations), their affect, and their visit experiences and communication with their physicians. Results showed (a) noteworthy disagreements between patients' self-reports and medical charts regarding cancer screening; (b) better agreement of patients' self-reports with videotape records than with chart records regarding physicians' recommendations; (c) accurate recognition of patients' self-reported affect, communication, and visit experiences by third-party raters of both audiotapes and videotapes; and (d) similar correlations of audio- and videotape ratings with patients' self reports as well as substantial correlations between audio and video ratings. The implications of these findings are discussed, and recommendations are made for future research. PMID- 14527866 TI - Silence and cancer: why do families and patients fail to communicate? AB - This study examined the phenomenon of avoidance of family communication about cancer. Thirty-seven Stage III or IV lung cancer patients and 40 caregivers, including 24 primary and 16 secondary caregivers, were interviewed; a total of 26 families were studied. The interviews were audiotaped and transcribed. Analysis of the interviews indicated that two thirds of the families (65%) experienced communication problems. The avoidance of family communication was associated with several underlying thought processes: avoidance of psychological distress; desire for "mutual protection;" and belief in positive thinking. Family communication was further hindered by the increasing difficulty of issues inherent to late stage cancer. The adverse impact of communication avoidance and the implications of our findings are discussed. PMID- 14527867 TI - Predictors of communication quality: the patient, provider, and nurse call center triad. AB - The purpose of this study was to examine characteristics of communication quality in a managed care context in which a nurse call center was used as the patient point of entry to the health system. The study sought to determine the level of communication quality among patients, health care providers, and nurses in the call center. Having measured the timeliness, accuracy, usefulness, and quantity of communication variables, a reasonable picture of communication quality emerged. The study explored patient differences in their perception of communication quality due to socioeconomic status, needs, experience, age, and various other factors. Likewise, providers who differ in training, specialty, and experience should have different perceptions and expectations of communication through nurse call center interaction. PMID- 14527868 TI - Are social norms campaigns really magic bullets? assessing the effects of students' misperceptions on drinking behavior. AB - In an attempt to curb excessive drinking on college campuses, many universities have turned to "social norms" marketing campaigns. Despite widespread acceptance among health educators, empirical results are clouded by measurement problems. This study, based on a random sample of 550 students, examined the effects of misperceptions of friends' and typical college students' drinking on one's drinking behavior. Results indicate that drinking behavior is positively related to perceptions of friends' drinking as suggested by the theory of planned behavior, which emphasizes subjective as opposed to social norms. PMID- 14527869 TI - Which adolescents are most receptive to tobacco industry marketing? implications for counter-advertising campaigns. AB - This study sought to identify adolescents most receptive to tobacco advertising based on individual differences in novelty-seeking personality and other key variables. Confidential self-report surveys were completed by 1,071 high school freshmen at 5 public high schools. The survey included validated measures of novelty-seeking personality, smoking habits, peer and family smoking, and tobacco advertising receptivity. Multiple logistic regression analysis was used to evaluate the independent associations of these variables and demographics with receptivity to tobacco advertising. Of the ninth graders, 44% had moderate to high levels of advertising receptivity and 54% had minimal to low levels of receptivity. Higher levels of receptivity were associated with ever smoking (odds ratio [OR] = 2.59, confidence interval [CI] = 1.99-3.39) and novelty-seeking personality (OR = 2.14, CI = 1.57-2.93). The association of novelty-seeking personality and tobacco advertising receptivity was most pronounced among adolescents who had never had a puff of a cigarette. Counter-advertising messages should consider individual differences in novelty-seeking, because novelty seekers may be most receptive to tobacco industry promotional campaigns. PMID- 14527870 TI - Experiences of physicians who frequently use e-mail with patients. AB - Despite its potential, the use of e-mail for physician-patient communication has not been widely adopted. Our purpose was to survey the experiences of physicians who are early adopters of the technology. Physicians, identified through a professional Internet information portal, completed a survey, including an assessment of satisfaction with using e-mail with patients. We identified 204 physicians who reported using e-mail with patients on a daily basis. Average age of the respondents was 49 years, 82% were male, and 35% were primary-care physicians. Among the 204 frequent users, commonly reported e-mail topics were new, nonurgent symptoms, and questions about lab results. Despite their daily use, 25% were not satisfied with physician-patient e-mail. The most important reasons for using e-mail with patients among those who were satisfied were "time saving" (33%) and "helps deliver better care" (28%) compared with "patient requested" (80%) among those who were not satisfied (p <.01). Dissatisfied physicians reported concerns about time demands, medicolegal risks, and ability of patients to use e-mail appropriately. Although the majority of these "vanguard" physicians reported benefits, some did not recommend that colleagues adopt this new technology. Increasing integration into practice to enhance time saving aspects and improve patient education might lead to more sustained use of this promising communication tool. PMID- 14527871 TI - The serotonin precursor 5-hydroxytryptophan delays neuromuscular disease in murine familial amyotrophic lateral sclerosis. AB - INTRODUCTION: Reduction in the levels of whole-blood serotonin is a common feature of Down syndrome (DS) individuals and transgenic mice overexpressing wild type SOD1. Administration of the metabolic precursor 5-hydroxytryptophan (5-HTP) leads to reversal of both serotonin deficits and hypotonia in humans. The effect of 5-HTP treatment on the progression of motor neuron disease in mutant SOD1 mice was examined. METHODS: Pre-disease transgenic SOD1 G93A mice and wild-type littermates were systemically administered 5-HTP thrice weekly (0, 5 or 50 mg/kg). Animal weights, locomotor function and survival were recorded weekly. Plasma serotonin levels were measured post-mortem. RESULTS: Treatment with 5-HTP significantly delayed hindlimb weakness and mortality in SOD1 G93A mice in a dose dependent manner. Wild-type mice were not adversely affected by 5-HTP administration. Baseline serotonin levels did not differ between wild-type and ALS mice. Blood platelet serotonin levels increased proportionally with dose. CONCLUSIONS: Increased blood serotonin by administration of 5-HTP in SOD1 G93A mice led to improved locomotor function and survival. A role for serotonin metabolism in mice with elevated SOD1 expression and motor neuron disease is suggested by these studies. PMID- 14527872 TI - Stable expression and characterization of human PN1 and PN3 sodium channels. AB - Nociceptive transduction in inflammatory and neuropathic pain involves peripherally expressed voltage-gated sodium channels, such as tetrodotoxin (TTX) sensitive PN1 and TTX-resistant PN3. We generated recombinant cell lines stably expressing the human PN1 and PN3 sodium channels in Chinese hamster ovary (CHO) cells using inducible expression vectors. The PN1 and PN3 cDNAs were isolated from human adrenal gland and heart poly(A)+ RNAs, respectively. The recombinant human PN1 currents exhibited rapid activation and inactivation kinetics and were blocked by TTX with a half-maximal inhibitory concentration (IC50) of 32.6 nM. The human PN3 channel expressed in stable transfectants showed TTX-resistant inward currents with slow inactivation kinetics. The IC50 value for TTX was 73.3 microM. The voltage-dependence of activation of the PN3 channel was shifted to the depolarizing direction, compared to that of the PN1 channel. Lidocaine and mexiletine exhibited tonic and use-dependent block of PN1 and PN3 channels. The PN1 channel was more susceptible to inhibition by mexiletine than PN3. These results suggest that stable transfectants expressing the human PN1 and PN3 sodium channels will be useful tools to define subtype selectivity for sodium channel blockers. PMID- 14527873 TI - Proteinous amino acids in muscle cytosol of rats' heart after exercise and hypoxia. AB - Levels of 19 proteinous amino acids and of total free amino acids were assayed by gas-liquid chromatography in cytosols of rat atrial and ventricular heart muscle cardiomyocytes. These amino acids were assayed after the rats had been exposed to either exercise (swimming) or hypoxia (hypobaric pressure of 686 hectoPascals). Out of the total free amino acids levels of arginine, glutamine and cysteine in atrial and ventricular cardiac muscle cytosols of control rats were the highest of all amino acids assayed. The control levels of all other amino acids assayed in atrial or ventricular cardiac muscles ranged from 0.1% to 10.6% of the total free amino acids in the control rats. Physical stress (exercise and hypoxia) significantly reduced the total amount of cytosolic free amino acids in both heart muscles. While hypoxia decreased the levels of arginine in both heart muscles, exercise abolished the level of cysteine in the atrial heart muscle. Decrease in arginine levels, and elimination of cysteine from the heart's atrial muscle after physical stress, may be attributed to its utilization of nitric oxide and to its synthesis of atriopeptin and/or endothelin during stress. No change was recorded in either experimental group in the level of glutamine in heart muscle cytosol. Exercise and hypoxia affect, in different modes, the levels of all other amino acids assayed, except for tryptophan, tyrosine, and histidine, which are precursors of endogenous neurotransmitters. The impact of proteinous amino acids on some bodily functions is discussed. PMID- 14527874 TI - A bifunctional alkylating nitrogen mustard agent that utilizes barbituric acid as carrier drug with the potential for crossing the brain-blood barrier. AB - Barbituric acid is the parent compound of a large family of hypnotic barbiturates. A nitrogen mustard (N-mustard) group (-CH2CH2N[CH2CH2Cl]2) was placed onto the two nitrogen atoms at positions 1 and 3 of the pyrimidine ring. This N-mustard agent is a solid at 25 degrees C, stable at -10 degrees C for >10 weeks, and soluble in aqueous solvent at 37 degrees C and 25 degrees C. The partition coefficients miLog P and CLog P were calculated to be -0.93 and -1.441 for barbituric acid. The miLog P and CLog P for the N-mustard agent were 1.82 and 2.707, respectively. The N-mustard substituents significantly increased solubility in lipid by-layers. The N-mustard agent alkylated a nucleophilic primary amine (p-chloroaniline) at physiological conditions of pH 7.4 and 37 degrees C. Aliquots of reaction mixtures were withdrawn at known time periods to react with fluorescamine for determination of unreacted p-chloroaniline and calculation of rate constants. The alkylation of the primary amine was second order with rate = k2[Nu]2, (Nu is nucleophile) and rate constant k2 = 0.01358 L/(mole.min). The molecular dipole of barbituric acid and the N-mustard agent was calculated by SPARTAN software (wavefunction, Irvine, CA) to be 0.681 and 2.153 Debye, respectively. The brain/blood partition coefficient (Log BB) of the N mustard agent was -0.399. Values of molecular polar surface area (TPSA) for barbituric acid and the N-mustard agent was 75.27 and 64.17, respectively. TPSA values indicate an expected intestinal absorbance to be 79% and 90%, respectively. The N-mustard agent showed zero violations of the Rule of 5, indicating good bioavailability. PMID- 14527875 TI - Properties of Arg389-beta1-adrenoceptor-Gsalpha fusion proteins: comparison with Gly389-beta1-adrenoceptor-Gsalpha fusion proteins. AB - The human beta1-adrenoceptor (beta1AR) exists in several isoforms and activates adenylyl cyclase (AC) via Gs-proteins. The Arg389-isoform of the beta1AR (beta1AR R389) expressed in CHW cells is much more efficient than the Gly389 isoform of the beta1AR (beta1AR-G389) at stabilizing the ternary complex and activating AC (Mason et al. 1999). The beta1AR-G389 fused to the Gsalpha splice variants GsalphaL or GsalphaS is efficient at stabilizing the ternary complex and activating AC (Wenzel-Seifert et al. 2002). Here, we show that beta1AR-R389 Gsalpha fusion proteins and beta1AR-G389-Gsalpha fusion proteins are similarly efficient at stabilizing the ternary complex and activating AC. In terms of agonist efficacies and agonist potencies in the [35S]guanosine 5'-O-(3 thiotriphosphate) binding assay, beta1AR-R389-Gsalpha fusion proteins and beta1AR G389-Gsalpha fusion proteins are similar, too. Our present data fit to an increasing number of clinical studies that failed to detect physiology- or pathology-related functional differences between beta1AR-R389 and beta1AR-G389. PMID- 14527876 TI - The effects of flumazenil on the antinociceptive actions of morphine in rats. AB - The 8-fluoro-5,6-dihydro-5-methyl-6-oxo-4H-imidazol[1,5-a][1,4]benzodiazepine-3 carboxylic acid ethyl ester (Flumazenil)-morphine interaction on analgesia (acute pain model, tail-flick test) was tested after intraperitoneal (IP) and intrathecal (IT) routes of administration in female rats. Analgesia was enhanced by the concurrent administration of Flumazenil with morphine (IP), in a dose related way. Flumazenil alone (IP) did not produce analgesia. In contrast, morphine analgesia was not enhanced by Flumazenil by the IT route. These data demonstrate that Flumazenil enhances morphine-mediated antinociception by mechanisms that are likely to involve benzodiazepine receptors at sites other than the spinal cord. PMID- 14527877 TI - Combined effects of Cordyceps sinensis and methotrexate on hematogenic lung metastasis in mice. AB - We investigated antitumor effects of water extracts of Cordyceps sinensis (WECS). WECS (100 microg/ml) induced apoptosis of B16 melanoma cells after 48 h exposure in vitro as determined by both the TUNEL (TdT-mediated dUTP-biotin nick end labeling) method and the detection of a DNA ladder. In vivo, combined treatment with WECS and methotrexate (MTX) in mice intravenously inoculated with B16 melanoma cells was conducted. Although MTX caused a significant and severe decrease in body weight compared with that in control mice starting 16 days after the start of administration, the mice given both MTX and WECS did not show a significant decrease in body weight. The mean survival time (days) of the control mice, MTX-treated mice (15 mg/kg/day, s.c.), and WECS-treated mice (200 mg/kg/day, p.o.) was 25.0 +/- 0.3, 25.6 +/- 1.3, and 25.7 +/- 1.0 (mean +/- S.E.M. of 6-7 mice), respectively. On the other hand, mean survival time (days) of mice given the combination of MTX and WECS was 28.2 +/- 0.7, significantly longer than the control value. WECS might be beneficial in the prevention of tumor metastasis as an adjuvant agent in cancer chemotherapy. PMID- 14527878 TI - Effects of (+/-)-idazoxan alone and in combination with L-DOPA methyl ester in MPTP-induced hemiparkinsonian monkeys. AB - The effects of a combination of the alpha2-adrenergic receptor antagonist (+/-) idazoxan with L-DOPA methyl ester were examined in three of four female adult monkeys (Macaca nemestrina) rendered hemiparkinsonian with 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine (MPTP). (+/-)-Idazoxan (0.16, 0.63, and 1.0 mg/kg) was given i.m. 10 min before L-DOPA methyl ester (12.5 mg/kg). (+/-)-Idazoxan reduced the maximum peak of contralateral circling elicited during the first hour following injection of L-DOPA methyl ester, but prolonged the duration of the circling response up to 50% (p < 0.05). The data support a role for alpha2 adrenergic receptor mechanisms in modulating the effects of L-DOPA on nigrostriatal dopamine function in the MPTP monkey model of hemiparkinsonism. PMID- 14527881 TI - The extraordinary value of solitude in the surgeon's life. PMID- 14527879 TI - Biogenic amines in striatum of rats that had been treated with ethanol, and their brains later stored in different temperatures. AB - The purpose of this study was to investigate how ethanol pretreatment and storage temperatures of brain striatum affect levels of biogenic amines in this tissue. Adult Wistar male rats were injected with 25% ethanol (5.0 g/kg i.p.) while the control rats were administered i.p. with the same volume of saline. Two hours later the rats were decapitated, their brains removed, and the striatum separated. Each striatum was divided into three parts: one part was immediately frozen on dry ice and kept at -70 degrees C; a second fragment was kept in a household refrigerator (+4 degrees C); and the third fragment was kept at +22 degrees C. Twenty-four hours later, levels of DA, DOPAC, HVA, 3-MT, 5-HT, and 5 HIAA in the striatum were assayed by HPLC/ED. Immediately after decapitation; ethanol levels were assayed in the serum of ethanol-pretreated and saline pretreated rats using gas chromatography. Our results indicate that levels of striatal DA, DOPAC, and HVA in saline-pretreated rats decreased significantly when the storage temperature of the striatum was raised from -70 degrees C, through +4 degrees C, to +22 degrees C, while levels of striatal 5-HT and 5-HIAA remained constant within the temperature range tested and levels of 3-MT fluctuated. In ethanol-pretreated rats, striatal levels of DOPAC, HVA, and 5-HIAA were increased in all three storage temperatures, while levels of DA, 5-HT, and 3 MT were decreased in those temperatures. Those decreases were most profound in striatal samples kept at +22 degrees C. We conclude that concern about possible interactions between drugs and biogenic amines should be exercised. PMID- 14527882 TI - Classics of modern surgery: the unknown man of Alexis Carrel-- father of transplantation. PMID- 14527884 TI - Effect of titanium surface texture on the cell-biomaterial interface. AB - To improve the in vivo performance of engineered implants, this study examines the independent effects of surface chemistry and topography on fibroblast morphology and density in vitro. Titanium (Ti) was sputter-coated onto smooth and microtextured polyethylene terephthalate (PET). Test specimens were evaluated in 24-h, fibroblast cultures and assessed with scanning electron microscopy (SEM) for cellular morphology and density. Fibroblast density increased, as the Ti film thickness increased. The fibroblasts exhibited contact guidance on the textured test specimens. The greatest cellular density was found on the Ti-coated, textured test specimens. In conclusion, Ti and surface texture appeared to strongly influence fibroblast density and morphology as compared to PET and smooth surfaces. PMID- 14527883 TI - The attenuation of hemorrhage-induced liver injury by exogenous nitric oxide, L arginine, and inhibition of inducible nitric oxide synthase. AB - We investigated the role of nitric oxide (NO) in its ability to reduce liver injury in an animal model of hemorrhagic shock (HS). Ninety-six Sprague-Dawley rats weighing 250 to 300 g were divided in 6 groups (n = 16 per group) that included treatment at the beginning of resuscitation with normal saline (groups 1, 3) sodium nitroprusside (NP) (0.5 mg/kg) (groups 2, 4) L-arginine (300 mg/kg) (group 5), and L-N6-(1-iminoethyl) lysine (L-NIL, 40 mg/kg) (group 6). The experimental model of HS consisted of the withdrawal of 3 mL blood per 100 g in a 15-min period, tail amputation (75%), and drug administration at 30 min. This was followed by fluid resuscitation (FR) with lactated Ringer's (LR) solution to reach a mean arterial pressure (MAP) of 40 mm Hg, then a hospital phase of 60 min with hemostasis and FR with LR solution to reach a MAP of 70 mm Hg with a 3-day observation phase. NP, L-Arginine, and L-NIL significantly reduced fluid requirements for resuscitation (p =.0001) as well as significantly increased MAP after resuscitation from hemorrhage. We also observed an improved statistically significant difference (p =.001) in tests demonstrating less hepatic injury and histology damage. The mRNA expression of cytokines in the liver (interleukin [IL] 1alpha, IL-beta1, tumor necrosis factor [TNF]beta, IL-3, IL-4, IL-5, IL-6, IL-10, TNFalpha, IL-2, interferon [IFN]gamma) was reduced by NP treatment, L-arginine, and L-NIL. These data suggest that excess NO mediates hemorrhage-induced liver injury and that the suppression of inducible nitric oxide synthase (iNOS) generated NO bioavailability with the NO donor sodium nitroprusside may reduce the pathophysiologic consequences of severe hemorrhage. This effect could be possibly related to the scavenging of to superoxide radicals (O2-) or the blockade of the deleterious effects of TNF and other inflammatory cytokines. The protective action noted with L-arginine cannot be fully explained within the context of this article, although it could be most likely associated with the supplementation of eNOS-generated NO. PMID- 14527885 TI - Hyperoxic condition prevents bacterial translocation and elevation of plasma microorganism components during hemorrhagic shock. AB - To evaluate the influence of hyperoxic conditions on bacterial translocation (BT) and microorganism components during hemorrhagic shock, rats were divided into a group breathing 100% oxygen and a group breathing room air. The groups were then subjected to hemorrhagic shock. Systemic blood and mesenteric lymph nodes were cultured for BT, and systemic plasma concentrations of microorganism components were measured by the silkworm larvae plasma (SLP) test and the endotoxin test. Hyperoxic conditions prevented both BT and plasma SLP-reactive substance (peptidoglycan and beta-glucan) elevation during hemorrhagic shock. Our findings suggest that hyperoxic treatment might improve host conditions during hemorrhagic shock. PMID- 14527886 TI - Prostaglandins E1 and E2 analogues ameliorate mucosal injury secondary to distal colonic obstruction. AB - After colonic obstruction, increased intraluminal pressure may impair blood circulation and cause mucosal injury that sometimes progresses to perforation. This experimental study aims to evaluated effects of prostaglandin E (PGE) analogues in order to prevent mucosal injury secondary to distal colonic obstruction. This study was carried out on 40 male Wistar rats equally divided into four groups: group 1, control; group 2, colonic obstruction by ligation of the sigmoid colon; group 3, obstruction and administration of PGE1; group 4, obstruction and administration of PGE 2. Mucosal structural changes were assessed and scored by histopathological examination at 24 h postoperatively. The thickness of the mucosal layer of the cecal wall was measured as 179.3 and 122 microm (p <.001) in the control and obstruction alone group, respectively. Mucosal thickness was preserved by PGE1 (170 microm) and PGE2 (157.3 microm) administration. The mean damage score was 0.73 in the control and 3.3 (p <.001) in the obstruction alone group. This score was significantly lower in obstruction groups treated with both PGE1 and PGE2, 1.13 and 1.26 respectively (p <.001). Both PGE1 and PGE2 (PGE1 better than PGE2) ameliorate mucosal injury during distal colonic obstruction. We concluded that the administration of PGE analogues in case of mechanical obstruction of the left colon was beneficial for preserving the structure of the mucosal layer. PMID- 14527887 TI - Electrophysiologic identification of the location of the colonic pacemakers in humans: further study. AB - We recently defined the sites of four colonic pacemakers that appear to generate the electric waves assumed to be responsible for the colonic motility. We hypothesized that a dysfunction of one or more of these pacemakers might interfere with the generation of electric waves and the colonic motility. This hypothesis was investigated in the current communication. The tests were performed during the repair of huge incisional hernia of 8 subjects (5 F, 3 M; mean age 42.8 +/- 3.3 SD years). Two electrodes were applied to each of the terminal ileum (TI), cecum (C), and ascending (AC), transverse (TC), descending (DC), and sigmoid (SC) colon. The electric activity of the TI and the various colonic segments was recorded using surface silver-silver chloride electrodes applied to the colon. The site of change of the wave variables between the TI and the C and between the different other colonic segments was determined by changing the position of the electrodes placed over the segments to be examined. Presumably, the sites where the wave variables changed represent the potential location of the pacemakers. We anesthetized these sites individually by injection of 2% Xylocaine, and then recorded the electric activity after 20 min in all the subjects and after 2 h in only 5 subjects. Electric waves in the form of pacesetter and action potentials were recorded from the TI and the colon. The sites of potential pacemakers could be defined at the ileocecal and cecocolonic junctions, at the mid third of the TC, and at the colosigmoid junction. Anesthetization of the cecal pole resulted in disappearance of the cecal electric waves, with persistence of the waves from the other colon segments. Anesthetization of the cecocolonic junction eliminated the electric waves of the AC and the right half of the TC, while the waves in the rest of the colon persisted. The remaining two pacemaker sites produced similar results when anesthetized. The electric waves reappeared after the anesthetic effect had waned. Thus, the colon possesses at least four pacemakers that appear to mediate the colonic motor activity. Individual pacemaker block by anesthetization effected disappearance of electric waves in the relative colonic segment, which reappeared after waning of the anesthetic effect. The disappearance of these waves upon pacemaker anesthetization supports a relationship between the pacemakers at the anesthetized site and the electric waves. The electric waves seem to be generated by these pacemakers. We suggest that colonic inertia, segmental or total, results from the dysfunction of one or more pacemakers, and that an artificial pacemaker could be applied for the treatment of such conditions. These suggestions need to be further studied. PMID- 14527888 TI - Prothrombotic disorders in patients with mesenteric vein thrombosis. AB - Mesenteric vein thrombosis (MVT) is uncommon condition. The purpose of this study was to assess prevalence of prothrombotic disorders in these patients. Eleven patients with MVT were screened for protein C, protein S, and antithrombin III deficiencies. Gene analysis was performed by polymerase chain reaction. A prothrombotic disorder was detected in 9 (81.8%) patients. Factor V Leiden, methylenetetrahydrofolate reductase TT677, and prothrombin G20210A mutations were found in 2 (18.2%), 2 (18.2%), and 5 patients (45.4%), respectively. Protein S, protein C, and antithrombin III deficiencies were present in 1, 1, and 2 patients, respectively. Four patients (36.3%) had combined defects. Thus, prothrombotic disorders may have a causative role in the pathogenesis of MVT, and these patients must be screened for these disorders. PMID- 14527890 TI - Immortalization of epididymal epithelium in transgenic mice expressing simian virus 40 T antigen: characterization of cell lines and regulation of the polyoma enhancer activator 3. AB - In the present study epididymal epithelium was immortalized in transgenic mice by expressing simian virus 40 T antigen under a 5.0-kb mouse glutathione peroxidase 5 promoter (GPX5-Tag1). Epididymal tumorigenesis was associated with an increase in c-Myc expression, and a marked decrease in B-Myc expression, with a 500-fold lower level in the GPX5-Tag1 caput epididymis compared with wild-type caput. Furthermore, B-Myc was undetectable in the immortalized corpus and cauda epididymis. Hence, it is possible that the normally high B-Myc expression in the epididymis is one of the factors contributing to the highly resistant nature of epididymis toward immortalization. Morphologically different epithelial cell lines were generated from the immortalized epididymides, and the cells expressed several genes typical for epididymal epithelium, such as mouse epididymal 1, mouse epididymal protein 9, androgen and estrogen receptors, anion exchangers 2 and 4, retinoic acid receptor alpha, and polyoma enhancer activator 3 (PEA3). This indicated the differentiated status of the cells and their usefulness for analyzing epididymal gene expression in vitro. As PEA3 is considered to be one of the transcription factors responsible for epididymal gene expression, we further studied its regulation in epididymal cells in vitro. The data showed that PEA3 mRNA expression is regulated in the epididymis via protein kinase A and ERK signaling cascades. Inhibiting protein kinase A resulted in up-regulation and inhibiting ERK resulted in down-regulation of PEA3 mRNA, whereas no significant effect on PEA3 expression was found by modulating the protein kinase C, stress activated p38, phosphoinositol 3-kinase and p70 S6 kinase cascades. PMID- 14527891 TI - Do human myeloma cells directly produce basic FGF? PMID- 14527892 TI - Hypereosinophilic syndrome with elevated serum tryptase versus systemic mast cell disease associated with eosinophilia: 2 distinct entities? PMID- 14527893 TI - Divergence from the germ-line sequence in unmutated chronic lymphocytic leukemia is due to somatic mutation rather than polymorphisms. PMID- 14527894 TI - Clinical research to standard of care: when does the transition occur? PMID- 14527895 TI - Management of peritoneal carcinomatosis: is an answer at hand? PMID- 14527896 TI - Colorectal cancer prevention studies: the importance of defining disease risk. PMID- 14527897 TI - Beyond p53. PMID- 14527898 TI - Gallbladder cancer gene hypermethylation: genetics or environment? PMID- 14527899 TI - Long-term disease-free survival after resection of hepatocellular carcinoma: both tumor behavior and surgeon's performance are important determinants. PMID- 14527900 TI - Quality assurance: the value of data and the will to improve. PMID- 14527901 TI - Recent insights into angiogenesis, apoptosis, invasion, and metastasis in colorectal carcinoma. AB - The numerous studies profiling mechanisms in colorectal carcinoma have implicated multiple pathways in the malignant progression of a colorectal epithelial cell. Such pathways as aberrations in the cell cycle, deviation from apoptosis, neovascularization of tumors, and invasion and metastasis of malignant epithelial cells have been shown to occur in the progression of a normal epithelial cell to an adenoma and carcinoma. Today, we continue to search for communications or connections between these pathways as we try to get a more global picture of the events responsible for the adenoma-carcinoma sequence. This review focuses on the latest developments of three well-characterized pathways implicated in colorectal carcinoma: angiogenesis, apoptosis, and invasion and metastasis. We will attempt to highlight clinical correlates, when available, with some of the more interesting molecules. PMID- 14527902 TI - Overcoming antiapoptotic responses to promote chemosensitivity in metastatic colorectal cancer to the liver. AB - BACKGROUND: Metastatic colon cancer is highly resistant to chemotherapy. A variety of mechanisms by which cancer cells resist chemotherapy have been described including enhanced export of drug from cancer cells and alterations in drug metabolism. In addition, the response of cancer cells to genotoxic therapies may be diminished by acquired defects in either the response mechanisms to DNA damage or cell cycle regulatory pathways. Recently, attention has focused on mechanisms that are activated by treatment exposure and subsequently promote resistance by rescuing cancer cells from apoptosis. The objective of this review is to examine the role of antiapoptotic mechanisms of chemotherapy resistance and to determine the potential utility of therapeutic strategies that target these mechanisms. METHODS: To accomplish the objectives, a brief overview of mechanisms of chemotherapy resistance is provided. The concept of inducible chemotherapy resistance is introduced by examination of a specific antiapoptotic mechanism, mediated by the transcription factor, nuclear factor kappa B (NF-kappa B). The ability to use inhibitors of NF-kappa B to promote chemosensitivity is examined in vitro and in vivo. RESULTS: Inhibition of chemotherapy-induced NF-kappa B activation enhances apoptosis and augments chemotherapy sensitivity. CONCLUSIONS: NF-kappa B inhibition may overcome cancer cell defense against apoptosis. Molecular therapies that target this resistance mechanism may be useful adjuncts to conventional chemotherapy. PMID- 14527903 TI - Intraperitoneal chemohyperthermia using a closed abdominal procedure and cytoreductive surgery for the treatment of peritoneal carcinomatosis: morbidity and mortality analysis of 216 consecutive procedures. AB - BACKGROUND: Peritoneal carcinomatosis has been regarded as a lethal clinical entity. Recently, aggressive treatments combining intraperitoneal chemohyperthermia (IPCH) with cytoreductive surgery have resulted in long-term survival in selected patients. The aim of this trial was to analyze the mortality and morbidity of 216 consecutive treatments of peritoneal carcinomatosis by IPCH by using a closed abdominal procedure combined with cytoreductive surgery. METHODS: Between February 1989 and August 2001, 207 patients who underwent 216 IPCH procedures using a closed abdominal procedure with mitomycin C, cisplatin, or both were prospectively studied. RESULTS: The postoperative mortality and morbidity rates were 3.2% and 24.5%, respectively. The most frequent complications were digestive fistula (6.5%) and hematological toxicity (4.6%). Morbidity was statistically linked with the carcinomatosis stage (P =.016), the duration of surgery (P =.005), and the number of resections and peritonectomy procedures (P =.042). Duration of surgery and carcinomatosis stage were the most common predictors of morbidity. CONCLUSIONS: The frequency of complications after IPCH and cytoreductive surgery was mainly associated with the carcinomatosis stage and the extent of the surgical procedure. The IPCH closed abdominal procedure has shown an acceptable frequency of adverse events. PMID- 14527904 TI - Adenoma recurrences after resection of colorectal carcinoma: results from the Southwest Oncology Group 9041 calcium chemoprevention pilot study. AB - BACKGROUND: Colorectal adenomas are the usual precursors to carcinoma in sporadic and hereditary colorectal cancers (CRC). METHODS: A total of 220 CRC patients (stages 0, I, and II) were randomized prospectively in a double-blind pilot study of calcium chemoprevention by using recurrent colorectal adenomas as a surrogate end point. This trial is still in progress, and we report the preliminary findings on adenoma recurrence rates. RESULTS: Synchronous adenomas were present in 60% of patients, and cancer confined in a polyp was present in 23% of patients. The overall cumulative adenoma recurrence rate was 31% (19% in the first year, 29% for 2 years, and 35% for 3 years). The recurrence rates were greater for patients with synchronous adenomas: 38% at 3 years (P =.01). Lower stage was associated with higher adenoma recurrence rates (P =.04). Factors including age, sex, site of primary cancer, and whether the cancer was confined to a polyp were not significantly associated with differences in adenoma recurrence rates. CONCLUSIONS: The substantial adenoma recurrence rate in patients resected of CRC justifies colonoscopic surveillance on a periodic basis. Patients with higher rates of adenoma recurrences, such as CRC with synchronous adenomas, are ideal subjects for chemoprevention trials. PMID- 14527905 TI - Intratumoral heterogeneity in microsatellite alterations in BRCA1 and PTEN regions in sporadic colorectal cancer. AB - BACKGROUND: Chromosome regions 17q21 (BRCA1) and 10q23 (PTEN) have been found deleted in colorectal cancer. METHODS: We studied the frequency of loss of heterozygosity (LOH) in these 2 regions in 214 patients with only 1 sample per tumor and in 100 patients with several samples per tumor. Three microsatellite markers of each region were used for the LOH test. The polymerase chain reaction product was electrophoresed in 8% polyacrylamide gels, and band intensity was shown by silver staining. RESULTS: The proportions of LOH in the two regions were 38.4% for 17q21 and 30.8% for 10q23 in the group of 214 and were 47.7% for 17q21 and 34.7% for 10q23 in the group of 100. We found a high correlation between the LOH in both regions (P <.001), where 81% of LOH in 10q23 region was matched by concomitant LOH in 17q21. In the group of tumors with several samples (group of 100), 39% and 68% did not present LOH in the 17q21 and 10q23 regions, respectively, in all of their tumor samples. However, in the 20 patients with LOH in both regions in the group of 100 (several samples per tumor), all samples with LOH in 10q23 also had LOH in 17q21, whereas not all samples with LOH in 17q21 had LOH in 10q23. CONCLUSIONS: These results show that colorectal cancer is highly heterogeneous, at least for these tumors markers, and suggest a sequential acquisition pattern of these anomalies during tumor growth, in which changes in 17q21 could occur before those in 10q23. PMID- 14527906 TI - Progression of gene hypermethylation in gallstone disease leading to gallbladder cancer. AB - BACKGROUND: Aberrant methylation of tumor-suppressor genes is associated with a loss of gene function that can afford selective growth advantages to sporadic neoplastic cells arising during gallbladder inflammation. METHODS: Fifty-four gallbladder neoplasms were selected from tumor banks in the United States and Chile. Each of the neoplasms was subjected to methylation-specific polymerase chain reaction to detect promoter methylation associated with six candidate tumor suppressor genes (p16, APC, methylguanine methyltransferase, hMLH1, retinoic acid receptor beta-2, and p73) implicated in multiple human cancer types. RESULTS: Aberrant methylation of any of the six candidate tumor-suppressor genes was detected in 72% of the gallbladder neoplasms, 28% of the cases of chronic cholecystitis, and in only 1 of the 15 normal gallbladder controls. The four most commonly methylated genes in the gallbladder cancers were p16 (56%), p73 (28%), APC (27%), and hMLH1 (14%). Significant differences in gene methylation were discovered between US gallbladder cancers and those from Chile, where gallbladder cancer is one of the leading causes of cancer-related deaths. APC methylation was present in 42% of the US cases but in only 14% of the Chilean tumors (P =.028). p73 methylation was common among the Chilean cancers (40%) compared with those from the United States (13%; P =.034). CONCLUSIONS: The acquisition of hypermethylation at multiple tumor-suppressor gene-promoter sites may contribute to tumor formation and progression within the chronically inflamed gallbladder. The apparent differences in methylation patterns among the Chilean and US gallbladder cases may indicate a unique biology associated with this cancer in different parts of the world. PMID- 14527907 TI - Comparison of linear array endoscopic ultrasound and helical computed tomography for the staging of periampullary malignancies. AB - BACKGROUND: The purpose of this study was to compare linear array endoscopic ultrasound (EUS) and helical computed tomography (CT) scan in the preoperative local staging evaluation of patients with periampullary tumors. METHODS: Patients evaluated with EUS and CT for suspected periampullary malignancies from 1996 to 2000 were analyzed. Surgical/pathology staging results were the reference standard. RESULTS: Forty-eight patients (28 men and 20 women; mean age, 62 +/- 4.9 years; range, 18-90 years) were identified. Malignancy was histologically confirmed in 44 patients. Parameters evaluated included tumor size, lymph node metastases, and major vascular invasion. EUS was significantly more sensitive (100%), specific (75%), and accurate (98%) than helical CT (68%, 50%, and 67%, respectively) for evaluation of the periampullary mass (P <.05). In addition, EUS detected regional lymph node metastases in more patients than helical CT. Sensitivity, specificity, and accuracy of EUS were 61%, 100%, and 84%, in comparison to 33%, 92%, and 68%, respectively, with CT. Major vascular involvement was noted in 9 of 44 patients. EUS correctly identified vascular involvement in 100% compared with 45% with CT (P <.05). CONCLUSIONS: Linear array EUS was consistently superior to helical CT in the preoperative local staging of periampullary malignancies. PMID- 14527908 TI - Follow-up surveillance for recurrence after curative gastric cancer surgery lacks survival benefit. AB - BACKGROUND: Although routine follow-up to detect asymptomatic recurrence after surgery for gastric cancer is recommended, the effect of such reassessment on survival has not been evaluated. METHODS: Clinical records of patients developing recurrent disease after potentially curative resection between 1985 and 1996 were retrieved. Among these patients, 197 were in our follow-up program. We analyzed survival in these patients according to the presence or absence of cancer-related symptoms when recurrent disease was diagnosed. RESULTS: Of all patients with recurrent disease, 50% were diagnosed within 1 year and 75% within 2 years of surgery. Asymptomatic recurrence, detected in 88 patients (45%), frequently represented distant metastasis. Although early detection significantly improved survival after detection of recurrent disease, disease-free survival for this subset was shorter. Thus, no significant difference in overall survival was observed. CONCLUSIONS: Early detection of asymptomatic gastric cancer recurrence did not improve overall survival of patients with recurrence after curative resection. Until development of more effective treatment for this disease, close follow-up may offer no survival benefit. PMID- 14527909 TI - Lymph node retrieval and assessment in stage II colorectal cancer: a population based study. AB - BACKGROUND: Adjuvant chemotherapy for patients with stage III (node-positive) colorectal cancer (CRC) reduces mortality by one third. Retrieval of an inadequate number of lymph nodes in the surgical specimen may result in incorrectly designating some patients as stage II (node negative), and consequently, such patients may not be offered appropriate chemotherapy. Recent National Cancer Institute guidelines suggest that a minimum of 12 nodes should be examined to ensure accurate staging. METHODS: This population-based study identified stage II (T3N0 and T4N0) CRC cases by using CRC pathology reports (1997-2000) from the Ontario Cancer Registry. Patients aged 19 to 75 years were identified, and demographic, surgical, pathologic, and hospital data were extracted. Factors relating to the number of lymph nodes assessed were examined. RESULTS: A total of 8848 CRC cases were reviewed, and 1789 stage II cases were identified. Seventy-three percent of cases were designated as node negative on the basis of assessment of <12 lymph nodes. Multivariate analysis showed that age, tumor size, specimen length, use of a pathology template, and academic status of the hospital were significant predictors of the number of lymph nodes assessed. CONCLUSIONS: A subset of patients with CRC in Ontario were assigned stage II disease on the basis of examination of relatively few lymph nodes. PMID- 14527910 TI - Telomerase inhibition using azidothymidine in the HT-29 colon cancer cell line. AB - BACKGROUND: We investigated the effects of telomerase inhibition by using the reverse transcriptase inhibitor azidothymidine (AZT) in the human colorectal cancer cell line HT-29 in the presence and absence of 5-fluorouracil (5-FU). METHODS: HT-29 cells were cultured in the presence of AZT. Telomerase activity was measured by using the telomerase repeat amplification protocol. Telomere length was determined by Southern analysis. The colorimetric microtiter assay was performed to determine the cytotoxic effects of AZT, alone and in combination with 5-FU. RESULTS: The presence of 3'-azido-3'-deoxythymidine triphosphate (AZT TP) effectively inhibited telomerase extracted from HT-29 cells. HT-29 cells cultured with 125 microM of AZT underwent fewer total population doublings over 91 days. Southern analysis revealed that telomere attrition occurred within this period. Exposure to 125 microM of AZT resulted in slightly reduced viability (10%) of HT-29 cells. However, the presence of AZT increased 5-FU cytotoxicity, suggesting that the effects of these two drugs are synergistic. CONCLUSIONS: The data are consistent with telomerase inhibition having growth-inhibitory effects in addition to those predicted to accompany loss of telomere function. Further studies using specific small-molecule inhibitors will confirm whether the growth inhibitory and 5-FU-sensitivity effects seen here are a direct result of telomerase inhibition. PMID- 14527911 TI - Predictors of long-term disease-free survival after resection of hepatocellular carcinoma: two decades of experience at Chang Gung Memorial Hospital. AB - BACKGROUND: Hepatocellular carcinoma (HCC) is a common disease in Taiwan. A high incidence of recurrence after hepatectomy is the most important cause of unsatisfactory results after resection of HCC. This study aimed to determine the clinicopathologic factors for predicting survival >5 years without recurrence in HCC patients treated with hepatectomy. METHODS: The clinical features of 46 patients with surgically resected HCC who survived >5 years without recurrence (group A) were reviewed. Also, comparison was made with the clinical features and factors influencing the outcome of 1046 cases of resected HCC who survived <5 years both with and without recurrence and for >5 years with recurrence (group B). RESULTS: Of 1092 cases of surgically resected HCC, 46 (4.5%) survived >5 years without recurrence. Univariate analysis revealed that absence of vascular invasion, satellite lesions, low histological grading of HCC, a lower rate of underlying liver cirrhosis, wider resection margins, and an uneventful postoperative course were frequent in group A patients compared with group B patients. Moreover, multivariate stepwise logistic regression analysis identified an absence of satellite lesions and an uneventful postoperative course as the two main independently significant predictors of HCC patients undergoing hepatectomy with long-term disease-free survival. The 1-, 3-, 5-, and 10-year survival rates of group A patients were 100%, 100%, 100%, and 84.1%, whereas those of group B patients were 73.5%, 47.9%, 29.0%, and 21.2%, respectively. CONCLUSIONS: An absence of satellite lesions and an uneventful postoperative course are the two main independent predictors for long-term disease-free survival in HCC patients undergoing hepatic resection. PMID- 14527912 TI - Pancreatic resection for metastatic renal cell carcinoma: presentation, treatment, and outcome. AB - BACKGROUND: The diverse natural history of renal cell carcinoma (RCC) includes metastases to the pancreas, a very unusual site for distant spread of other cancers. Considering the relatively indolent behavior of some cases of metastatic RCC, pancreatic resection is offered to select patients. METHODS: We reviewed the records of patients at three affiliated university hospital centers who had prior nephrectomy for RCC and subsequent pancreatic resection of metastases. RESULTS: Fourteen patients--9 women and 5 men with a median age of 63.8 years--underwent a total of 15 pancreatic resections for metastatic RCC. Nine (64%) had solitary metastases. The median interval from nephrectomy to diagnosis of pancreatic metastases was 83 months. The median size of metastases was 4.6 cm. There was one perioperative death. Pancreatic recurrence occurred in five patients (36%), and one patient underwent repeat resection. At a median follow-up of 32 months, seven patients (50%) are alive without evidence of disease, and four patients (28%) are alive with recurrent disease. CONCLUSIONS: Resection of pancreatic metastases from RCC is associated with long-term survival and should be considered for patients in whom complete resection is possible. PMID- 14527913 TI - Intra-arterial infusion chemotherapy with angiotensin-II for locally advanced and nonresectable pancreatic adenocarcinoma: further evaluation and prognostic implications. AB - BACKGROUND: For locally advanced and nonresectable cancer of the pancreas, we performed intra-arterial infusion chemotherapy with angiotensin-II (AT-II). In our preliminary report, this treatment resulted in a median of 14 months of survival without objective adverse effects. This study was designed to clarify the prognostic factor in this chemotherapy by using a larger number of cases. METHODS: For 32 patients, intra-arterial chemotherapy was performed: 1 or 2 catheters were intraoperatively placed into the pancreas-supplying arteries. The tissue blood flow and its change by AT-II infusion were determined. For intra arterial chemotherapy, a mixture of methotrexate (50 or 100 mg/m(2)) and AT-II (.4 microg/kg/hour) was repeatedly infused from the catheter, mainly at our outpatient clinic. RESULTS: With our intra-arterial chemotherapy, the median survival period was 13 months. The median survival period was 19 months in patients without coexisting pancreatitis but was only 9 months in those with it (P =.0003). The presence or absence of coexisting fibrosis in the neighboring uninvolved pancreas offered the only prognostic indicator. The blood flow in cancerous tissue was increased during AT-II infusion, and this was characteristic in the patients whose neighboring uninvolved pancreas had normal parenchyma (nonatrophic) or higher blood flow before AT-II infusion. CONCLUSIONS: Because the AT-II infusion played a role in shifting the blood flow from the surrounding uninvolved pancreas to the cancer tissues, we can speculate that cancer tissues might have thereby received a higher dose of anticancer drugs if the surrounding uninvolved pancreas had been nonfibrotic and more rich in tissue blood flow. PMID- 14527914 TI - Should the hunt for internal mammary chain sentinel nodes begin? An evaluation of 150 breast cancer patients. AB - BACKGROUND: The aim of this study was to determine the visualization rate, identification rate, and clinical implications of biopsy of sentinel nodes in the internal mammary chain (IMC) in patients with breast cancer. METHODS: From January 1999 to December 2002, 691 sentinel node procedures were performed. Preoperative lymphoscintigraphy was performed after injection of (99m)Tc-labeled nanocolloid into the tumor (.2 mL; 115 MBq; 3.1 mCi). The sentinel node was surgically identified with the aid of patent blue dye and a gamma ray detection probe. RESULTS: The sentinel node in the IMC could be harvested in 130 (87%) of the 150 patients in whom it was visualized on the images and contained metastases in 22 (17%) of these 130 cases. In nine patients (7%), the IMC sentinel node was tumor positive, whereas the axilla was tumor-free. Stage migration was seen in all patients with a tumor-positive IMC sentinel node (17%). There was a change of management in 38 (29%) of the 130 patients: institution or omission of radiotherapy to the IMC, adjuvant systemic therapy, or omission of an axillary lymph node dissection. CONCLUSIONS: Pursuit of IMC sentinel nodes improves the staging of patients with breast cancer and enables treatment to be better adjusted to the needs of the individual patient. PMID- 14527915 TI - Subareolar injection may be more accurate than other techniques for sentinel lymph node biopsy in breast cancer. AB - BACKGROUND: The aims of this study were to compare peritumoral injection of (99m)Tc-labeled albumin and subdermal injection of blue dye with subareolar (SA) injection of blue dye alone in terms of success of the sentinel lymph node identification rate, false negative (FN) rate, overall accuracy, and sensitivity of the two procedures. METHODS: From January 1999 to October 2002, 155 patients with localized breast cancer were treated. Patients were subdivided into two groups. In patients in group 1 (n = 115; January 1999 to December 2001), lymphoscintigraphy together with injection of vital dye was performed. In patients in group 2 (n = 40; January 2002 to October 2002), SA injection of blue dye alone was performed. RESULTS: In patients in group 1, the overall successful identification rate was 94.8%. The success rate of identifying a sentinel lymph node by a combination of the two techniques was 95%. With blue dye alone, the successful identification rate was 94.6% in patients in group 1 (subdermal) and 97.5% in group 2 (SA). The FN rate was 9% in group 1 and 0% in group 2. The overall accuracy of lymphatic mapping was 97% in group 1 and 100% in group 2. Sensitivity was 91% in group 1 and 100% in group 2. CONCLUSIONS: This study of dye-only injection into the SA plexus demonstrates a high sentinel node identification rate, absent FN rate, and rapid learning curve. On the basis of these findings, we propose that injections into the SA lymphatic plexus are the optimal way to perform dye-only lymphatic mapping of the breast. PMID- 14527916 TI - Nipple aspirate fluid expression of urokinase-type plasminogen activator, plasminogen activator inhibitor-1, and urokinase-type plasminogen activator receptor predicts breast cancer diagnosis and advanced disease. AB - BACKGROUND: Tumor expression of urokinase-type plasminogen activator (uPA), plasminogen activator inhibitor-1 (PAI-1), and uPA receptor (uPAR) are breast cancer prognostic factors. Less is known about their usefulness in breast cancer diagnosis. Nipple aspirate fluid (NAF) is secreted into the breast duct and collected noninvasively, making it potentially useful both in breast cancer diagnosis and prognosis. We determined the association of uPA, PAI-1, and uPAR levels in NAF with breast cancer (1) detection and (2) advanced disease. METHODS: A total of 88 NAF specimens were collected from women with or without breast cancer, and uPA, PAI-1, and uPAR expression were measured by enzyme-linked immunosorbent assay. RESULTS: uPA and uPAR were independent predictors of cancer presence; uPAR was also an independent predictor of advanced disease stage. Higher PAI-1 expression in breast cancer that was found with univariate analysis was not observed after logistic regression was applied. CONCLUSIONS: NAF evaluation of uPA, uPAR, and, perhaps, PAI-1 (significant only in univariate analysis) may provide useful breast cancer diagnostic and prognostic information. PMID- 14527917 TI - Prospective analysis of accuracy of positron emission tomography, computed tomography, and endoscopic ultrasonography in staging of adenocarcinoma of the esophagus and the esophagogastric junction. AB - BACKGROUND: Exact preoperative staging of esophageal cancer is essential for accurate prognosis and selection of appropriate treatment modalities. METHODS: Forty-two patients with adenocarcinoma of the esophagus or the esophagogastric junction suitable for radical esophageal resection were staged with positron emission tomography (PET), spiral computed tomography (CT), and endoscopic ultrasonography (EUS). RESULTS: Diagnostic sensitivity for the primary tumor was 83% for PET and 67% for CT; for local peritumoral lymph node metastasis, it was 37% for PET and 89% for EUS; and for distant metastasis, it was 47% for PET and 33% for CT. Diagnostic specificity for local lymph node metastasis was 100% with PET and 54% with EUS, and for distant metastasis, it was 89% for PET and 96% for CT. Accuracy for locoregional lymph node metastasis was 63% for PET, 66% for CT, and 75% for EUS, and for distant metastasis, it was 74% with PET and 74% with CT. Of the 10 patients who were considered inoperable during surgery, PET identified 7 and CT 4. The false-negative diagnoses of stage IV disease in PET were peritoneal carcinomatosis in two patients, abdominal para-aortic cancer growth in one, metastatic lymph nodes by the celiac artery in four, and metastases in the pancreas in one. PET showed false-positive lymph nodes at the jugulum in three patients. CONCLUSIONS: The diagnostic value of PET in the staging of adenocarcinoma of the esophagus and the esophagogastric junction is limited because of low accuracy in staging of paratumoral and distant lymph nodes. PET does, however, seem to detect organ metastases better than CT. PMID- 14527918 TI - Buttock soft tissue sarcoma: clinical features, treatment, and prognosis. AB - BACKGROUND: Primary buttock soft tissue sarcomas in adults are common entities that have been infrequently reported (three clinical series and isolated case reports). We present our experience of buttock sarcomas to better characterize and define the natural history of this condition. METHODS: Buttock tumors occurring in adults (>16 years) between January 1990 and January 2002 were identified from the Royal Marsden Hospital's Sarcoma Unit prospective database. RESULTS: Seventy-three buttock sarcomas were evaluated and treated at the Royal Marsden Hospital during this period. Liposarcoma (n = 19), leiomyosarcoma (n = 13), and synovial sarcoma (n = 9) were the most frequent histological types. There were 8 T1 and 61 T2 tumors, and size was not available in 4 patients. Most tumors (n = 64) were located deep to the deep fascia. There were 15 grade 1, 20 grade 2, and 37 grade 3 tumors, and grade was not available in 1 patient. There were 29 tumors contained within the gluteus maximus. Wide excision was performed in 50 patients. Local recurrence and distant metastasis occurred in 15 and 35 patients with a median time of 18 and 8 months, respectively. The rate of local recurrence at 2 years was 20.9% (SE, 6.8%). The 2-year overall and disease-free survival rates were 64.1% (SE, 6.7%) and 48.5% (SE, 6.4%), respectively. CONCLUSIONS: Buttock sarcomas present special surgical difficulties because of proximity of the sciatic nerve and the ability of tumors at this site to extend into the pelvis and perineum. Size and grade of the tumor were independent predictors for disease-free and overall survival. PMID- 14527919 TI - Potential prophylactic measures against postoperative immunosuppression: could they reduce recurrence rates in oncological patients? AB - BACKGROUND: Removing the primary tumor is indispensable for eliminating the major pool of metastasizing cells, but the surgical procedure itself is suspected of promoting metastases. This adverse effect is attributed to several mechanisms acting in synergy, including mechanical release of tumor cells, enhanced angiogenesis, secretion of growth factors, and immunosuppression. Here we provide new insights into mechanisms of postoperative immunosuppression and assess the assumptions underlying the hypothesis that, by suppressing cell-mediated immunity (CMI), surgery may render the patient vulnerable to metastases that otherwise could have been controlled. METHODS: An extensive review of relevant articles in English identified by using the MEDLINE database and cross-referencing. RESULTS: Current literature suggests that (1) CMI can control minimal residual disease, especially if surgery is performed early; (2) major surgery transiently but markedly suppresses CMI through multiple mechanisms now better understood; (3) surgical stress promotes experimental metastasis through immunosuppression, but the clinical evidence remains indirect because of ethical limitations. CONCLUSIONS: Minimizing postoperative immunosuppression seems feasible, may limit recurrence, and should be introduced into the broader array of considerations when planning oncological surgeries. In the short run, physicians could try to avoid immunosuppressive anesthetic approaches, inadvertent hypothermia, excessive blood transfusions, and untended postoperative pain. When feasible, minimally invasive surgery should be considered. In the long run, clinical trials should evaluate prophylactic measures, including perioperative immunostimulation and several antagonists to cytokines and hormones specified herein. PMID- 14527921 TI - Predicting nonsentinel lymph node involvement in stage I/II melanoma. PMID- 14527922 TI - Breast conservation for T1-2 breast carcinomas. PMID- 14527924 TI - Airway epithelial integrity is protected by a long-acting beta2-adrenergic receptor agonist. AB - Airway epithelial integrity may be impaired by bacterial exoproducts, which are able to degrade tight junction-associated proteins such as zonula occludens 1 (ZO 1). We have investigated the protective effect of salmeterol, a long-acting beta(2)-adrenergic agonist, on Pseudomonas aeruginosa-induced alteration of the epithelial junctional barrier. We demonstrate in human airway epithelial cells (HAEC) that salmeterol induces a time-dependent increase in ZO-1 protein, although no significant change in ZO-1 transcripts was observed. When HAEC cultures were exposed to P. aeruginosa (PAO1) supernatants, apical expression of ZO-1 protein was maintained in salmeterol-pretreated HAEC cultures, whereas it disappeared after PAO1 exposure in cultures not pretreated with salmeterol. Western blot experiments showed that the 220-kD ZO-1 protein was decreased after PAO1 incubation but was still present in salmeterol-pretreated HAEC extracts. The functional activity of ZO-1 protein was monitored by measuring transepithelial resistance and analyzing the diffusion of a low molecular weight tracer through the intercellular spaces. After PAO1 incubation, the epithelial integrity of HAEC was impaired, as shown by a decrease in transepithelial resistance and increased paracellular permeability, but was not significantly altered after salmeterol preincubation. These results demonstrate that salmeterol may contribute to the protection of the airway epithelium barrier against bacterial virulence factors. PMID- 14527925 TI - p53 N-terminal Ser-15 approximately P and Ser-20 approximately P levels in squamous cell lung cancer after radio/chemotherapy. AB - Functional regulation of p53 protein, a critical regulator of cell cycle and apoptosis, was investigated in fiberoptic bronchoscopy biopsy samples taken from 23 patients suffering from recurrent squamous cell lung cancer by analyzing the expression and phosphorylation status of the p53 at Ser15 and Ser20 before and after treatment with radiotherapy/cisplatin/vinorelbine. Poly(ADP-ribose) levels as a marker of cellular DNA damage, expression of wild-type and mutated p53 protein, and Ki-67 expression as a marker of proliferation was also determined. Median p53 expression increased (61% increase) after therapy. p53 phosphorylated on Ser20 was also increased by approximately 57% in radiotherapy/chemotherapy patients, and these changes correlated with Ki-67 proliferation and with elevated (by 69%; P < 0.01) poly(ADP-ribose) levels. Our data indicate that apart from changes in p53 quantity, post-translational phosphorylation/dephosphorylation mediated alterations, especially at Ser20 may play a role in p53 stabilization and, therefore, in antiproliferative activity of drugs inducing DNA damage and apoptosis. PMID- 14527926 TI - Resident murine alveolar and peritoneal macrophages differ in adhesion of apoptotic thymocytes. AB - Apoptotic cells must be cleared efficiently by macrophages (Mo) to prevent autoimmunity, yet their ingestion impairs Mo microbicidal function. The principal murine resident lung phagocyte, the alveolar Mo (AMo), is specifically deficient at apoptotic cell ingestion, both in vitro and in vivo, compared with resident peritoneal Mo (PMo). To further characterize this deficiency, we assayed static adhesion in vitro using apoptotic thymocytes and resident AMo and PMo from normal C57BL/6 mice. Adhesion of apoptotic thymocytes by both types of Mo was rapid, specific, and cold-sensitive. Antibody against the receptor tyrosine kinase MerTK (Tyro12) blocked phagocytosis but not adhesion in both types of Mo. Surfactant protein A increased adhesion and phagocytosis by AMo, but not to the levels seen using PMo. Adhesion was largely cation-independent for PMo and calcium-dependent for AMo. Adhesion was not inhibited in either Mo type by mAbs against beta1 or beta3 integrins or scavenger receptor I/II (CD204), but AMo adhesion was inhibited by specific mAbs against CD11c/CD18. Thus, resident murine tissue Mo from different tissues depend on qualitatively disparate receptor systems to bind apoptotic cells. The decreased capacity of murine AMo to ingest apoptotic cells is only partially explained by reduced initial adhesion. PMID- 14527927 TI - Glucocorticoid inhibition of granulocyte macrophage-colony-stimulating factor from T cells is independent of control by nuclear factor-kappaB and conserved lymphokine element 0. AB - Release of granulocyte macrophage-colony-stimulating factor (GM-CSF) from T cells is important in the differentiation, maturation, and survival of inflammatory cells. Here the induction of GM-CSF expression from T cells was dependent on transcription and translation and was prevented by dexamethasone. In primary human CD3(+) T cells, up to 3.3 kb of human GM-CSF promoter was strongly activated by PMA + PHA. Mutations in either the -85/-76 nuclear factor (NF) kappaB site or the activator protein-1 region in the -54/-31 conserved lymphokine element 0 (CLE0) site substantially reduced promoter activity. Both GM-CSF promoter and NF-kappaB-dependent constructs were unresponsive to dexamethasone whereas the release of GM-CSF was potently repressed. Analysis of GM-CSF mRNA and protein expression at various time points and the effect of adding dexamethasone after the stimulus revealed the existence of potent mechanisms of inhibition acting at a translational level. The expression of tristetraproline and HuR, proteins that bind the AU-rich element in the GM-CSF 3'-untranslated region was unaffected by dexamethasone and overall AU-rich element binding activity was unaltered. Taken together our data support an important role for the NF-kappaB and CLE0 sites in the transcriptional control of GM-CSF expression in primary human T cells and suggest that post-transcriptional/translational mechanisms are key mediators of glucocorticoid-dependent repression. PMID- 14527928 TI - Post-BMT lung injury occurs independently of the expression of CCL2 or its receptor, CCR2, on host cells. AB - Idiopathic pneumonia syndrome (IPS) is a significant cause of mortality post-bone marrow transplant (BMT) in humans. In our murine model, lethal pre-BMT conditioning and allogeneic T cells result in the recruitment of host antigen presenting cells (APC) and donor T cells into the lung post-BMT concomitant with development of severe lung dysfunction. CCL2 induction is found in bronchoalveolar lavage fluid (BALF) before host monocyte influx. The major receptor for CCL2 is CCR2 present on monocytes; this interaction can play a crucial role in monocyte recruitment in inflammation. To determine whether blockade of the CCL2/CCR2 pathway could hinder host monocyte influx, lethally conditioned wild-type (WT), CCL2(-/-), or CCR2(-/-) mice were transplanted with allogeneic marrow and spleen cells. WT and (-/-) recipients exhibited equivalent lung dysfunction post-BMT. The frequencies of host macrophages as well as donor CD4(+) and CD8(+) T cells in lungs post-BMT did not differ between WT and (-/-) recipients. However, the T cell dependency of the host CD11b(+) major histocompatibility complex class II(+) cell influx was lost in CCR2(-/-) recipients. In CCR2(-/-) mice, this influx was accompanied by elevated levels of CCL20. Post-BMT BALF and sera of (-/-) mice did not reveal any decrease in cytokines or chemokines compared with WT mice. CCL2(-/-) mice had a deficiency of CCL2 in their BALF and sera post-BMT, confirming our hypothesis that CCL2 is predominantly host derived. Therefore, IPS can occur independently of host expression of CCL2 or CCR2, and compensatory mechanisms exist for regulating APC recruitment into the lung during the early post-BMT period. PMID- 14527929 TI - DNA synthesis and Bcl-2 expression during development of mucous cell metaplasia in airway epithelium of rats exposed to LPS. AB - Exposure of pulmonary airways to environmental toxins and allergens may cause proliferation of airway epithelial cells and mucous cell metaplasia (MCM); however, it is unclear to what extent proliferating cells differentiate into mucus-storing cells and contribute to MCM. Our previous studies demonstrated that Bcl-2, an inhibitor of apoptosis with cell cycle regulatory functions, is expressed in metaplastic mucous cells. The purpose of the present study was to investigate the number of metaplastic mucous cells that are derived from proliferating epithelial cells and whether Bcl-2 has a role in cell cycle entry in these cells. Rats were intratracheally instilled with 100 microg of LPS from Pseudomonas aeruginosa in 500 microl of saline, and proliferating airway cells were labeled with bromodeoxyuridine (BrdU) by implanting a subcutaneous osmotic pump 24 h before instillation. The volume of stored mucosubstance and the number of mucous cells were increased 10- and 3-fold, respectively, from 24-48 h after instillation. The number of total epithelial cells per millimeter of basal lamina increased, and the number of serous cells per millimeter of basal lamina decreased during this time. Approximately 50% of Alcian blue-periodic acid Schiff stained mucous cells were labeled with BrdU at 48 h after instillation, suggesting that one-half of the secretory cells were derived from proliferating cells. Furthermore, 50% of the Bcl-2-positive mucous cells were BrdU negative and therefore derived from nonproliferating, preexisting cells. Our findings demonstrate that preexisting and proliferating cells differentiate into mucous cells and compose LPS-induced metaplasia and that Bcl-2 does not have cell cycle regulatory function in these cells. PMID- 14527930 TI - Mitochondrial cytochrome c release is a key event in hyperoxia-induced lung injury: protection by cyclosporin A. AB - Hyperoxia is known to induce extensive alveolar cell death by still poorly defined mechanisms. In this study, the mitochondria-dependent cell death pathway was explored during hyperoxia-induced lung injury in mice. We observed a progressive release of cytochrome c from the mitochondria into the cytosol of alveolar cells. This release was accompanied by the translocation of the proapoptotic protein Bax from cytosol to mitochondria without detectable activation of caspase-3. As cytochrome c release can be induced by mitochondrial membrane alteration and permeability transition (MPT), mice were treated with cyclosporin A, which specifically inhibits MPT. Cyclosporin A treatment prevented mitochondrial release of cytochrome c during hyperoxia and concomitantly preserved mitochondria from extensive swelling and crista disorganization, as assessed by electron microscopy analysis of alveolar epithelial cells. These morphological and biochemical observations correlated with decreased lung tissue damage, as evaluated by morphological score and lung weight. In conclusion, mitochondrial damage and cytochrome c release are important linked events in hyperoxia-induced lung injury and can be efficiently blocked by cyclosporin A. PMID- 14527931 TI - Continuous subcutaneous injection reduces polymorphonuclear leukocyte activation by granulocyte colony-stimulating factor. AB - The use of granulocyte colony stimulating factor (G-CSF) for recovery from neutropenia has been established; however, acute lung injury due to G-CSF-induced polymorphonuclear leukocyte (PMN) activation is a serious complication. This study was designed to compare the activation of PMN with single bolus administration and continuous administration of G-CSF. Healthy volunteers (age 33.8 +/- 1.4 yr; n = 6) received a single bolus injection of 50 microm/m2 of G CSF (SI; n = 6) or continuous subcutaneous injection of 50 microm/m2 of G-CSF for 24 h (CI; n = 6) and were followed for 48 h. Circulating leukocyte counts, markers of activation on PMN, and circulating levels of G-CSF, IL-6, and PMN elastase were measured. SI rapidly increased serum G-CSF levels, which peaked at 4 h, whereas CI gradually increased G-CSF levels, which remained at a steady level from 8 to 24 h. SI caused a rapid decrease in PMN counts at 0.5 h followed by sustained increase to peak at 12 h. CI gradually increased PMN counts, which peaked at 24 h, but the peak values were not significantly different between the groups. SI-induced activation of PMN, which was characterized by increased expression of CD11b, decreased expression of L-selectin, and increased F-actin content, led to increases in serum IL-6 and PMN elastase level. Such changes were all attenuated with CI (P < 0.05). We conclude that continuous subcutaneous injection of G-CSF resulted in a marrow response similar to that to a single injection but yielded reduced PMN activation. PMID- 14527932 TI - Environmental oxygen tension affects phenotype in cultured bone marrow-derived macrophages. AB - This study tested the hypothesis that the unique phenotype of alveolar macrophages (AM) is maintained through adaptation to the relatively high oxygen partial pressure (P(O2)) of the lung, through modification of redox-sensitive transcription factors. BALB/c mouse bone marrow-derived macrophages (BMC) were differentiated under different P(O2) and compared functionally to AM and peritoneal macrophages (PM). BMC differentiated in normoxia (P(O2) 140 Torr, BMC(high)) were similar to AM in having low phagocytic and antigen presenting cell (APC) activities. However, BMC grown in low oxygen tension as found in other tissues (<40 Torr, BMC(low)) were better phagocytes and APCs, similar to PM. BMC(high) were more oxidative intracellularly than BMC(low), based on oxidation of dichlorofluorescein and higher glutathione disulfide/glutathione (GSH) ratios, despite having more GSH. Finally, lipopolysaccharide-induced nuclear factor kappaB translocation, measured by laser scanning cytometry, was reduced in BMC(high) and AM, compared with BMC(low) and PM, respectively. These data suggest that regulation of the AM phenotype may occur, at least in part, via inhibition of NF-kappaB by the unique redox environment. PMID- 14527933 TI - Regulation of MUC5AC mucin secretion and airway surface liquid metabolism by IL 1beta in human bronchial epithelia. AB - Mucociliary transport in the airways significantly depends on the liquid and mucin components of the airway surface liquid (ASL). The regulation of ASL water and mucin content during pathological conditions is not well understood. We hypothesized that airway epithelial mucin production and liquid transport are regulated in response to inflammatory stimuli and tested this hypothesis by investigating the effects of the pleiotropic, early-response cytokine, IL-1beta, on cultured primary human bronchial epithelial and second-passage, normal human tracheo-bronchial epithelial (NHTBE) cell cultures. Fully differentiated NHTBE cultures secreted two major airway mucins, MUC5AC and MUC5B. IL-1beta, in a dose- and time-dependent manner, increased the secretion of MUC5AC, but not MUC5B. MUC5AC mRNA levels were only transiently increased at 1 and 4 h after the start of IL-1beta treatment and returned to control levels thereafter, even though MUC5AC mucin production remained elevated for at least 72 h. Synchronous with elevated MUC5AC secretion, ASL volume increased, its percentage of solid was reduced, and the pH/[HCO(3)(-)] of the ASL was elevated. ASL volume changes reflected altered ion transport, including an upregulation of Cl(-) secretory currents (via CFTR and Ca(2+)-activated Cl(-) conductance) and an inhibition of epithelial sodium channel (ENaC)-mediated absorptive Na(+) currents. IL-1beta increased CFTR mRNA levels without affecting those for ENaC subunits. The synchronous regulation of ASL mucin and liquid metabolism triggered by IL-1beta may be an important defense mechanism of the airway epithelium to enhance mucociliary clearance during airway inflammation. PMID- 14527934 TI - Polymorphism in gene coding for ACE determines different development of myocardial fibrosis in rats. AB - In humans, the effect of angiotensin-converting enzyme (ACE) gene polymorphisms in cardiovascular disease is still controversial. In the rat, a microsatellite marker in the ACE gene allows differentiation of the ACE gene polymorphism among strains with different ACE levels. We tested the hypothesis that this ACE gene polymorphism determines the extent of cardiac fibrosis induced by isoproterenol (Iso) in the rat. We used a male F(2) generation (homozygous LL and BB ACE genotypes determined by polymerase chain reaction) derived from two rat strains [Brown-Norway (BB) and Lewis (LL)] that differ with respect to their plasma ACE activities. For induction of left ventricular (LV) hypertrophy (LVH) and cardiac fibrosis, rats were infused with Iso (5 mg x kg(-1) x day(-1)) or saline (control) for 10 days and euthanized at day 1 after the last injection. The interstitial collagen volumetric fraction (ICVF), collagen I, and fibronectin content, but not collagen III content, were significantly higher in the homozygous BB rats than in homozygous LL rats. Differences in metalloprotease (MMP)-9, but not in MMP-2 activities as well as in cardiac cell proliferation, were also detected between LL and BB rats treated with Iso. LV ACE activity was higher in BB rats than LL rats and correlated with ICVF (r = 0.61, P < 0.002). No changes were observed in plasma ACE activities, ANG II plasma or LV levels, plasma renin activity, and ACE and ANG II type 1 receptor (AT1R) mRNA levels in the LV of rats with the two different ACE polymorphisms. Iso induced a similar degree of LVH [assessed by an increase in LV weight 100 per body weight, LV-to right ventricle (RV) ratio, and LV protein content] in LL and BB rats. We concluded that rats in the F(2) generation with high plasma ACE activity developed more fibrosis but to a similar degree of LVH compared with rats with low plasma ACE activity. PMID- 14527935 TI - Effect of high-salt diet on NO release and superoxide production in rat aorta. AB - Sprague-Dawley rats were fed either a high-salt (HS) diet (4.0% NaCl) or a low salt (LS) diet (0.4% NaCl) for 3 days. Nitric oxide (NO) and superoxide production were assessed in the thoracic aorta by evaluating the fluorescence signal intensity from 4,5-diaminofluorescein (DAF-2DA) and dihydroethidine, respectively. Methacholine caused increased NO release in the aortas from rats on a LS but not HS diet. The SOD mimetic tempol restored methacholine-induced NO release in aortas from rats on a HS diet. Methacholine also caused superoxide production in the aortas of rats on a HS diet but not in the aortas of rats on a LS diet. Tempol and N(G)-monomethyl-l-arginine eliminated methacholine-induced superoxide production in the aortas of rats on a HS diet. Aortic rings from rats on the HS diet showed impaired methacholine-induced relaxation, which was improved by tempol. Tempol alone caused a NO-dependent relaxation of norepinephrine-precontracted aortas that was significantly greater in the aortas of rats on the HS diet than in vessels from rats on the LS diet. These data suggest that a HS diet impairs endothelium-dependent relaxation via reduced NO levels and increased superoxide production. PMID- 14527936 TI - A transmural pressure gradient induces mechanical and biological adaptive responses in endothelial cells. AB - A sudden increase in the transmural pressure gradient across endothelial monolayers reduces hydraulic conductivity (L(p)), a phenomenon known as the sealing effect. To further characterize this endothelial adaptive response, we measured bovine aortic endothelial cell (BAEC) permeability to albumin and 70-kDa dextran, L(p), and the solvent-drag reflection coefficients (sigma) during the sealing process. The diffusional permeability coefficients for albumin (1.33 +/- 0.18 x 10(-6) cm/s) and dextran (0.60 +/- 0.16 x 10(-6) cm/s) were measured before pressure application. The effective permeabilities (measured when solvent drag contributes to solute transport) of albumin and dextran (P(ealb) and P(edex)) were measured after the application of a 10 cmH(2)O pressure gradient; during the first 2 h of pressure application, P(ealb), P(edex), and L(p) were significantly reduced by 2.0 +/- 0.3-, 2.1 +/- 0.3-, and 3.7 +/- 0.3-fold, respectively. Immunostaining of the tight junction (TJ) protein zonula occludens 1 (ZO-1) was significantly increased at cell-cell contacts after the application of transmural pressure. Cytochalasin D treatment significantly elevated transport but did not inhibit the adaptive response, whereas colchicine treatment had no effect on diffusive permeability but inhibited the adaptive response. Neither cytoskeletal inhibitor altered sigma despite significantly elevating both L(p) and effective permeability. Our data suggest that BAECs actively adapt to elevated transmural pressure by mobilizing ZO-1 to intercellular junctions via microtubules. A mechanical (passive) component of the sealing effect appears to reduce the size of a small pore system that allows the transport of water but not dextran or albumin. Furthermore, the structures of the TJ determine transport rates but do not define the selectivity of the monolayer to solutes (sigma). PMID- 14527937 TI - P2 receptor-mediated Ca2+ transients in rat cerebral artery smooth muscle cells. AB - Significant Ca(2+) release was previously noted with the activation of L-type Ca(2+) current in rat superior cerebral artery smooth muscle cells. Here we examined whether the P(2X) current that is partly carried by Ca(2+) also triggers Ca(2+) release in this preparation. Application of P(2X) agonists evoked membrane currents and concomitant Ca(2+) transients in whole cell voltage-clamped single cells. The expected increase in intracellular Ca(2+) concentration ([Ca(2+)](i)) was calculated from the time-integrated P(2X) current by assuming Ca(2+) is the only charge carrier. The measured increase in [Ca(2+)](i) was plotted as a function of the expected increase in [Ca(2+)](i), and Ca(2+)-buffering power was obtained as a reciprocal of the linear fit to this relationship. Both ryanodine, a Ca(2+)-induced Ca(2+)-release inhibitor, and cADP ribose, a putative activator of Ca(2+)-induced Ca(2+) release, had no significant effects on Ca(2+)-buffering power. These results suggest that Ca(2+) influx through P(2X) receptors does not trigger significant Ca(2+) release. We then examined whether P(2X) responses influence the subsequent P(2Y) response. P(2Y) responses were characterized by measuring the rate of [Ca(2+)](i) increase obtained as the slope of the linear regression to the rising phase of the Ca(2+) transient. During simultaneous application of the P(2X) and P(2Y) agonist, the rate of [Ca(2+)](i) increase was facilitated or suppressed depending on the size of the P(2X) receptor-mediated [Ca(2+)](i) increase. Membrane depolarization close to the Ca(2+) equilibrium potential significantly promoted the rate of [Ca(2+)](i) increase. Our results suggest that the [Ca(2+)](i) increase and membrane depolarization caused by the P(2X) current may regulate the subsequent P(2Y) response. PMID- 14527938 TI - Contractile reserve but not tension is reduced in monocrotaline-induced right ventricular hypertrophy. AB - The objective of this study was to evaluate the role of right ventricular hypertrophy on developed tension (F(dev)) and contractile reserve of rat papillary muscle by using a model of monocrotaline (Mct)-induced pulmonary hypertension. Calcium handling and the influence of bicarbonate (HCO(3)(-)) were also addressed with the use of two different buffers (HCO(3)(-) and HEPES). Wistar rats were injected with either Mct (40 mg/kg sc) or vehicle control (Con). Isometrically contracting right ventricular papillary muscles were studied at 80% of the length of maximal developed force. Contractile reserve (1 - F(dev)/F(max)) was calculated from F(dev) and maximal tension (F(max)). Calcium recirculation was determined with postextrasystolic potentiation. Both groups of muscles were superfused with either HCO(3)(-) (Con-B and Mct-B, both n = 6) or HEPES (Con-H and Mct-H, both n = 6) buffer. With hypertrophy, contractions were slower but F(dev) was not changed. However, F(max) was decreased (P < 0.05). With HCO(3)(-), F(max) decreased from 23.8 +/- 6.5 mN.mm(-2) in Con-B, to 13.7 +/- 3.3 mN.mm(-2) in Mct-B. With HEPES, it decreased from 16.3 +/- 3.5 mN.mm(-2) (n = 6, Con-H) to 8.3 +/- 1.6 mN.mm(-2) (Mct-H). Contractile reserve during hypertrophy was therefore also decreased (P < 0.05). With HCO(3)(-), it decreased from 0.73 +/- 0.03 (Con-B) to 0.55 +/- 0.04 (Mct-B). With HEPES, it decreased (P < 0.001) from 0.64 +/- 0.07 (Con-H) to 0.19 +/- 0.06 (Mct-H). The recirculation fraction decreased (P < 0.05) from 0.59 +/- 0.04 in Con-B to 0.44 +/- 0.04 in Mct-B. We conclude that contractile reserve and recirculation fraction are impaired during hypertrophy, with a stronger effect under HEPES than HCO(3)(-) superfusion. PMID- 14527939 TI - Selective elimination of I(K,slow1) in mouse ventricular myocytes expressing a dominant negative Kv1.5alpha subunit. AB - Although previous studies have revealed a role for the voltage-gated K+ channel alpha-subunit Kv1.5 (KCNA5) in the generation of the 4-aminopyridine (4-AP) sensitive component of delayed rectification in mouse ventricles (IK,slow1), the phenotypic consequences of manipulating IK,slow1 expression in vivo in different (mouse) models are distinct. In these experiments, point mutations were introduced in the pore region of Kv1.5 to change the tryptophan (W) at position 461 to phenylalanine (F) to produce a nonconducting subunit, Kv1.5W461F, that is shown to function as a Kv1 subfamily-specific dominant negative (Kv1.5DN). With the use of the alpha-myosin heavy chain promoter to direct cardiac-specific expression, three lines of Kv1.5DN-expressing (C57BL6) transgenic mice were generated and characterized. Electrophysiological recordings from Kv1.5-DN expressing left ventricular myocytes revealed that the micromolar 4-AP sensitive IK,slow1 is selectively eliminated. The attenuation of IK,slow1 is accompanied by increased ventricular action potential durations and marked QT prolongation. In contrast to previous findings in mice expressing a truncated (DN) Kv1.1 transgene; however, no electrical remodeling is evident in Kv1.5DN-expressing ventricular myocytes, and the (Kv1.5DN-induced) elimination of IK,slow1 does not result in spontaneous ventricular arrhythmias. PMID- 14527940 TI - Phenotypic differences in transient outward K+ current of human and canine ventricular myocytes: insights into molecular composition of ventricular Ito. AB - The Ca(2+)-independent transient outward K(+) current (I(to)) plays an important electrophysiological role in normal and diseased hearts. However, its contribution to ventricular repolarization remains controversial because of differences in its phenotypic expression and function across species. The dog, a frequently used model of human cardiac disease, exhibits altered functional expression of I(to). To better understand the relevance of electrical remodeling in dogs to humans, we studied the phenotypic differences in ventricular I(to) of both species with electrophysiological, pharmacological, and protein-chemical techniques. Several notable distinctions were elucidated, including slower current decay, more rapid recovery from inactivation, and a depolarizing shift of steady-state inactivation in human vs. canine I(to). Whereas recovery from inactivation of human I(to) followed a monoexponential time course, canine I(to) recovered with biexponential kinetics. Pharmacological sensitivity to flecainide was markedly greater in human than canine I(to), and exposure to oxidative stress did not alter the inactivation kinetics of I(to) in either species. Western blot analysis revealed immunoreactive bands specific for Kv4.3, Kv1.4, and Kv channel interacting protein (KChIP)2 in dog and human, but with notable differences in band sizes across species. We report for the first time major variations in phenotypic properties of human and canine ventricular I(to) despite the presence of the same subunit proteins in both species. These data suggest that differences in electrophysiological and pharmacological properties of I(to) between humans and dogs are not caused by differential expression of the K channel subunit genes thought to encode I(to), but rather may arise from differences in molecular structure and/or posttranslational modification of these subunits. PMID- 14527941 TI - Effects of electroporation on optically recorded transmembrane potential responses to high-intensity electrical shocks. AB - The outcome of defibrillation shocks is determined by the nonlinear transmembrane potential (DeltaVm) response induced by a strong external electrical field in cardiac cells. We investigated the contribution of electroporation to DeltaVm transients during high-intensity shocks using optical mapping. Rectangular and ramp stimuli (10-20 ms) of different polarities and intensities were applied to the rabbit heart epicardium during the plateau phase of the action potential (AP). DeltaVm were optically recorded under a custom 6-mm-diameter electrode using a voltage-sensitive dye. A gradual increase of cathodal and well as anodal stimulus strength was associated with 1) saturation and subsequent reduction of DeltaVm; 2) postshock diastolic resting potential (RP) elevation; and 3) postshock AP amplitude (APA) reduction. Weak stimuli induced a monotonic DeltaVm response and did not affect the RP level. Strong shocks produced a nonmonotonic DeltaVm response and caused RP elevation and a reduction of postshock APA. The maximum positive and maximum negative DeltaVm were recorded at 170 +/- 20 mA/cm2 for cathodal stimuli and at 240 +/- 30 mA/cm2 for anodal stimuli, respectively (means +/- SE, n = 8, P = 0.003). RP elevation reached 10% of APA at a stimulus strength of 320 +/- 40 mA/cm2 for both polarities. Strong ramp stimuli (20 ms, 600 mA/cm2) induced a nonmonotonic DeltaVm response, reaching the same largest positive and negative values as for rectangular shocks. The transition from monotonic to nonmonotonic morphology correlates with RP elevation and APA reduction, which is consistent with cell membrane electroporation. Strong shocks resulted in propidium iodide uptake, suggesting sarcolemma electroporation. In conclusion, electroporation is a likely explanation of the saturation and nonmonotonic nature of cellular responses reported for strong electric stimuli. PMID- 14527942 TI - Hemodynamics of orthostatic intolerance: implications for gender differences. AB - Women have a greater incidence of orthostatic intolerance than men. We hypothesized that this difference is related to hemodynamic effects on regulation of cardiac filling rather than to reduced responsiveness of vascular resistance during orthostatic stress. We constructed Frank-Starling curves from pulmonary capillary wedge pressure (PCWP), stroke volume (SV), and stroke index (SI) during lower body negative pressure (LBNP) and saline infusion in 10 healthy young women and 13 men. Orthostatic tolerance was determined by progressive LBNP to presyncope. LBNP tolerance was significantly lower in women than in men (626.8 +/ 55.0 vs. 927.7 +/- 53.0 mmHg x min, P < 0.01). Women had steeper maximal slopes of Starling curves than men whether expressed as SV (12.5 +/- 2.0 vs. 7.1 +/- 1.5 ml/mmHg, P < 0.05) or normalized as SI (6.31 +/- 0.8 vs. 4.29 +/- 0.6 ml.m-2.mmHg 1, P < 0.05). During progressive LBNP, PCWP dropped quickly at low levels, and reached a plateau at high levels of LBNP near presyncope in all subjects. SV was 35% and SI was 29% lower in women at presyncope (both P < 0.05). Coincident with the smaller SV, women had higher heart rates but similar mean arterial pressures compared with men at presyncope. Vascular resistance and plasma norepinephrine concentration were similar between genders. We conclude that lower orthostatic tolerance in women is associated with decreased cardiac filling rather than reduced responsiveness of vascular resistance during orthostatic challenges. Thus cardiac mechanics and Frank-Starling relationship may be important mechanisms underlying the gender difference in orthostatic tolerance. PMID- 14527943 TI - Aspects, mechanism, and biological relevance of mitochondrial protein nitration sustained by mitochondrial nitric oxide synthase. AB - The goal of this study was to explore the occurrence of nitrated proteins in mitochondria given that these organelles are endowed with a mitochondrial nitric oxide (NO.-) synthase and considering the important role that mitochondria have in energy metabolism. Our hypothesis is that nitration of proteins constitutes a posttranslational modification by which NO.- exhibits long-term effects above and beyond those bioregulatory ones mediated through the interaction with cytochrome c oxidase. Our studies are aimed at understanding the mechanisms underlying the nitration of proteins in mitochondria and the biological significance of such a process in the cellular milieu. On promoting a sustained NO.- production by mitochondria, we investigated various aspects of protein nitration. Among them, the localization of nitrated proteins in mitochondrial subfractions, the identification of nitrated proteins through proteomic approaches, the characterization of affected pathways, and depiction of a target sequence. The biological relevance was analyzed by considering the turnover of native and nitrated proteins. In this regard, mitochondrial dysfunction, ensuing nitrative stress, may be envisioned as the result of accumulation of nitrated proteins, resulting from an overproduction of endogenous NO.- (this study), a failure in the proteolytic system to catabolize modified proteins, or a combination of both. Finally, this study allows one to gain understanding on the mechanism and nitrating species underlying mitochondrial protein nitration. PMID- 14527944 TI - Wolframin expression induces novel ion channel activity in endoplasmic reticulum membranes and increases intracellular calcium. AB - Wolfram syndrome is an autosomal recessive neuro-degenerative disorder associated with juvenile onset non-autoimmune diabetes mellitus and progressive optic atrophy. The disease has been attributed to mutations in the WFS1 gene, which codes for a protein predicted to possess 9-10 transmembrane segments. Little is known concerning the function of the WFS1 protein (wolframin). Endoglycosidase H digestion, immunocytochemistry, and subcellular fractionation studies all indicated that wolframin is localized to the endoplasmic reticulum in rat brain hippocampus and rat pancreatic islet beta-cells, and after ectopic expression in Xenopus oocytes. Reconstitution of wolframin from oocyte membranes into planar lipid bilayers demonstrated that the protein induced a large cation-selective ion channel that was blocked by Mg2+ or Ca2+. Inositol triphosphate was capable of activating channels in the fused bilayers that were similar to channel components induced by wolframin expression. Expression of wolframin also increased cytosolic calcium levels in oocytes. Wolframin thus appears to be important in the regulation of intracellular Ca2+ homeostasis. Disruption of this function may place cells at risk to suffer inappropriate death decisions, thus accounting for the progressive beta-cell loss and neuronal degeneration associated with the disease. PMID- 14527945 TI - Control of the Bacillus subtilis antiterminator protein GlcT by phosphorylation. Elucidation of the phosphorylation chain leading to inactivation of GlcT. AB - Bacillus subtilis transports glucose by the phosphotransferase system (PTS). The genes for this system are encoded in the ptsGHI operon, which is induced by glucose and depends on a termination/antitermination mechanism involving a riboswitch and the RNA-binding antitermination protein GlcT. In the absence of glucose, GlcT is inactive, and a terminator is formed in the leader region of the ptsG mRNA. If glucose is present, GlcT can bind to its RNA target and prevent transcription termination. The GlcT protein is composed of three domains, an N terminal RNA binding domain and two PTS regulation domains, PTS regulation domain (PRD) I and PRD-II. In this work, we demonstrate that GlcT can be phosphorylated by two PTS proteins, HPr and the glucose-specific enzyme II (EIIGlc). HPr dependent phosphorylation occurs on PRD-II and has a slight stimulatory effect on GlcT activity. In contrast, EIIGlc phosphorylates the PRD-I of GlcT, and this phosphorylation inactivates GlcT. This latter phosphorylation event links the availability of glucose to the expression of the ptsGHI operon via the phosphorylation state of EIIGlc and GlcT. This is the first in vitro demonstration of a direct phosphorylation of an antiterminator of the BglG family by the corresponding PTS permease. PMID- 14527946 TI - Key residues responsible for acyl carrier protein and beta-ketoacyl-acyl carrier protein reductase (FabG) interaction. AB - Fatty acid synthesis in bacteria is catalyzed by a set of individual enzymes collectively known as type II fatty-acid synthase. Each enzyme interacts with acyl carrier protein (ACP), which shuttles the pathway intermediates between the proteins. The type II enzymes do not possess primary sequence similarity that defines a common ACP-binding site, but rather are hypothesized to possess an electropositive/hydrophobic surface feature that interacts with the electronegative/hydrophobic residues along helix alpha2 of ACP (Zhang, Y.-M., Marrakchi, H., White, S. W., and Rock, C. O. (2003) J. Lipid Res. 44, 1-10). We tested this hypothesis by mutating two surface residues, Arg-129 and Arg-172, located in a hydrophobic patch adjacent to the active site entrance on beta ketoacyl-ACP reductase (FabG). Enzymatic analysis showed that the mutant enzymes were compromised in their ability to utilize ACP thioester substrates but were fully active in assays with a substrate analog. Direct binding assays and competitive inhibition experiments showed that the FabG mutant proteins had reduced affinities for ACP. Chemical shift perturbation protein NMR experiments showed that FabG-ACP interactions occurred along the length of ACP helix alpha2 and extended into the adjacent loop-2 region to involve Ile-54. These data confirm a role for the highly conserved electronegative/hydrophobic residues along ACP helix alpha2 in recognizing a constellation of Arg residues embedded in a hydrophobic patch on the surface of its partner enzymes, and reveal a role for the loop-2 region in the conformational change associated with ACP binding. The specific FabG-ACP interactions involve the most conserved ACP residues, which accounts for the ability of ACPs and the type II proteins from different species to function interchangeably. PMID- 14527947 TI - Oral fibroblast expression of wound-inducible transcript 3.0 (wit3.0) accelerates the collagen gel contraction in vitro. AB - Wounds of the oral mucosa show faster closure with less scar formation than skin wounds in other areas. A differentially expressed cDNA, wound-inducible transcript 3.0 (wit3.0), was isolated from oral mucosal wound in rats (Sukotjo, C., Abanmy, A. A., Ogawa, T., and Nishimura, I. (2002) J. Dent. Res. 81, 229 235). The purpose of this study was to characterize the wit3.0 gene structure and the function of its deduced peptide. Human and rat genome databases revealed that the gene for wit3.0 was located in human chromosome 12p11.23 and rat chromosome 4q44. Its human and rat gene structures were well conserved, composed of 7 exons spread over 20 kb. Exon 5 was alternatively spliced generating two transcripts encoding deduced peptides of 215 and 253 amino acids (wit3.0 alpha and wit3.0 beta, respectively). The protein families data base of alignments (Pfam) analysis suggested the wit3.0 peptide sequence shared similarity with a portion of the myosin II coiled-coil domain consensus sequence. Fibroblasts isolated from the rat oral wound up-regulated wit3.0 expression and exhibited greater ability to contract collagen gel in vitro than fibroblasts isolated from untreated oral mucosa/gingiva. NIH3T3 and rat oral fibroblasts transfected with expression vector containing the coding sequences of wit3.0 alpha or wit3.0 beta increased in vitro collagen gel contraction. When treated with TGF beta-1, NIH3T3 fibroblast expression of wit3.0 showed no significant change, whereas alpha smooth muscle actin was increased in a dose-dependent manner. These data suggest that there may be a novel wound healing pathway involving wit3.0 underlying the favorable early wound closure characteristics of oral mucosa. PMID- 14527948 TI - Lipase-selective functional domains of perilipin A differentially regulate constitutive and protein kinase A-stimulated lipolysis. AB - Perilipin (Peri) A is a lipid droplet-associated phosphoprotein that acts dually as a suppressor of basal (constitutive) lipolysis and as an enhancer of cyclic AMP-dependent protein kinase (PKA)-stimulated lipolysis by both hormone-sensitive lipase (HSL) and non-HSL(s). To identify domains of Peri A that mediate these multiple actions, we introduced adenoviruses expressing truncated or mutated Peri A and HSL into NIH 3T3 fibroblasts lacking endogenous perilipins and HSL but overexpressing acyl-CoA synthetase 1 and fatty acid transporter 1. We identified two lipase-selective functional domains: 1) Peri A (amino acids 1-300), which inhibits basal lipolysis and promotes PKA-stimulated lipolysis by HSL, and 2) Peri A (amino acids 301-517), which inhibits basal lipolysis by non-HSL and promotes PKA-stimulated lipolysis by both HSL and non-HSL. PKA site mutagenesis revealed that PKA-stimulated lipolysis by HSL requires phosphorylation of one or more sites within Peri 1-300 (Ser81, Ser222, and Ser276). PKA-stimulated lipolysis by non-HSL additionally requires phosphorylation of one or more PKA sites within Peri 301-517 (Ser433, Ser492, and Ser517). Peri 301-517 promoted PKA stimulated lipolysis by HSL yet did not block HSL-mediated basal lipolysis, indicating that an additional region(s) within Peri 301-517 promotes hormone stimulated lipolysis by HSL. These results suggest a model of Peri A function in which 1) lipase-specific "barrier" domains block basal lipolysis by HSL and non HSL, 2) differential PKA site phosphorylation allows PKA-stimulated lipolysis by HSL and non-HSL, respectively, and 3) additional domains within Peri A further facilitate PKA-stimulated lipolysis, again with lipase selectivity. PMID- 14527949 TI - Cell surface expression of GluR5 kainate receptors is regulated by an endoplasmic reticulum retention signal. AB - Kainate receptors (KARs) are mediators of excitatory neurotransmission in the mammalian central nervous system, and their efficient targeting and trafficking is critical for normal synaptic function. A key step in the delivery of KARs to the neuronal plasma membrane is the exit of newly assembled receptors from the endoplasmic reticulum (ER). Here we report the identification of a novel ER retention signal in the alternatively spliced C-terminal domain of the GluR5-2b subunit, which controls receptor trafficking in both heterologous cells and neurons. The ER retention motif consists of a critical arginine (Arg-896) and surrounding amino acids, disruption of which promotes ER exit and surface expression of the receptors, as well as altering their physiological properties. The Arg-896-mediated ER retention of GluR5 is regulated by a mutation that mimics phosphorylation of Thr-898, but not by PDZ interactions. Furthermore, two positively charged residues (Arg-900 and Lys-901) in the C terminus were also found to regulate ER export of the receptors. Taken together, our results identify novel trafficking signals in the C-terminal domain of GluR5-2b and demonstrate that alternative splicing is an important mechanism regulating KAR function. PMID- 14527950 TI - Generation of the beta-amyloid peptide and the amyloid precursor protein C terminal fragment gamma are potentiated by FE65L1. AB - Members of the FE65 family of adaptor proteins, FE65, FE65L1, and FE65L2, bind the C-terminal region of the amyloid precursor protein (APP). Overexpression of FE65 and FE65L1 was previously reported to increase the levels of alpha-secretase derived APP (APPs alpha). Increased beta-amyloid (A beta) generation was also observed in cells showing the FE65-dependent increase in APPs alpha. To understand the mechanism for the observed increase in both A beta and APPs alpha given that alpha-secretase cleavage of a single APP molecule precludes A beta generation, we examined the effects of FE65L1 overexpression on APP C-terminal fragments (APP CTFs). Our data show that FE65L1 potentiates gamma-secretase processing of APP CTFs, including the amyloidogenic CTF C99, accounting for the ability of FE65L1 to increase generation of APP C-terminal domain and A beta 40. The FE65L1 modulation of these processing events requires binding of FE65L1 to APP and APP CTFs and is not because of a direct effect on gamma-secretase activity, because Notch intracellular domain generation is not altered by FE65L1. Furthermore, enhanced APP CTF processing can be detected in early endosome vesicles but not in endoplasmic reticulum or Golgi membranes, suggesting that the effects of FE65L1 occur at or near the plasma membrane. Finally, although FE65L1 increases APP C-terminal domain production, it does not mediate the APP-dependent transcriptional activation observed with FE65. PMID- 14527951 TI - FoxO3a transcriptional regulation of Bim controls apoptosis in paclitaxel-treated breast cancer cell lines. AB - Paclitaxel is used to treat breast cancers, but the mechanisms by which it induces apoptosis are poorly understood. Consequently, we have studied the role of the FoxO transcription factors in determining cellular response to paclitaxel. Western blotting revealed that in a panel of nine breast cancer cell lines expression of FoxO1a and FoxO3a correlated with the expression of the pro apoptotic FoxO target Bim, which was associated with paclitaxel-induced apoptosis. In MCF-7 cells, which were paclitaxel-sensitive, the already high basal levels of FoxO3a and Bim protein increased dramatically after drug treatment, as did Bim mRNA, which correlated with apoptosis induction. This was not observed in MDA-231 cells, which expressed low levels of FoxOs and Bim. Gene reporter experiments demonstrated that in MCF-7 cells maximal induction of Bim promoter was dependent on a FoxO binding site, suggesting that FoxO3a is responsible for the transcriptional up-regulation of Bim. Gene silencing experiments showed that small interference RNA (siRNA) specific for FoxO3a reduced the levels of FoxO3a and Bim protein as well as inhibited apoptosis in paclitaxel-treated MCF-7 cells. Furthermore, siRNA specific for Bim reduced the levels of Bim protein and inhibited apoptosis in paclitaxel-treated MCF-7 cells. This is the first demonstration that up-regulation of FoxO3a by paclitaxel can result in increased levels of Bim mRNA and protein, which can be a direct cause of apoptosis in breast cancer cells. PMID- 14527952 TI - Modification of promyelocytic leukemia zinc finger protein (PLZF) by SUMO-1 conjugation regulates its transcriptional repressor activity. AB - Promyelocytic leukemia zinc finger (PLZF) protein is a sequence-specific DNA binding protein that represses the transcriptional activity of target genes such as those for cyclin A and the interleukin-3 receptor alpha chain. The PLZF gene becomes fused to the retinoic acid receptor alpha gene as a result of the t(11, 17)(q23;q21) chromosomal translocation that is associated with acute promyelocytic leukemia. We now show that endogenous PLZF in human promyelocytic leukemia HL-60 cells is modified by conjugation with SUMO-1 (small ubiquitin related modifier-1) and that PLZF colocalizes with SUMO-1 in the nucleus of transfected human embryonic kidney 293T cells. Site-directed mutagenesis identified lysine 242 in the RD2 domain of human PLZF as the sumoylation site. A luciferase reporter gene assay suggested that SUMO-1 modification of this residue is required for transcriptional repression by PLZF, and an electrophoretic mobility shift assay showed that this modification increases the DNA binding activity of PLZF. PLZF-mediated regulation of the cell cycle and transcriptional repression of the cyclin A2 gene were also dependent on sumoylation of PLZF on lysine 242. These results demonstrate that PLZF is modified by SUMO-1 conjugation and that this modification regulates the biological functions of PLZF. PMID- 14527953 TI - Substrate activation of insulin-degrading enzyme (insulysin). A potential target for drug development. AB - The rate of the insulin-degrading enzyme (IDE)-catalyzed hydrolysis of the fluorogenic substrate 2-aminobenzoyl-GGFLRKHGQ-ethylenediamine-2,4-dinitrophenyl is increased 2-7-fold by other peptide substrates but not by peptide non substrates. This increased rate is attributed to a decrease in Km with little effect on Vmax. An approximately 2.5-fold increase in the rate of amyloid beta peptide hydrolysis is produced by dynorphin B-9. However, with insulin as substrate, dynorphin B-9 is inhibitory. Immunoprecipitation of differentially tagged IDE and gel filtration analysis were used to show that IDE exists as a mixture of dimers and tetramers. The equilibrium between dimer and tetramer is concentration-dependent, with the dimer the more active form. Bradykinin shifted the equilibrium toward dimer. Activation of substrate hydrolysis is not seen with a mixed dimer of IDE containing one active subunit and one subunit that is catalytically inactive and deficient in substrate binding. On the other hand, a mixed dimer containing one active subunit and one subunit that is catalytically inactive but binds substrate with normal affinity is activated by peptides. These findings suggest that peptides bind to one subunit of IDE and induce a conformational change that shifts the equilibrium to the more active dimer as well as activates the adjacent subunit. The selective activation of IDE toward amyloid beta peptide relative to insulin suggests the potential for development of compounds that increase IDE activity toward amyloid beta peptide as a therapeutic intervention for the treatment of Alzheimer's disease. PMID- 14527954 TI - Overexpression of L-isoaspartate O-methyltransferase in Escherichia coli increases heat shock survival by a mechanism independent of methyltransferase activity. AB - Over time and under stressing conditions proteins are susceptible to a variety of spontaneous covalent modifications. One of the more commonly occurring types of protein damage is deamidation; the conversion of asparagines into aspartyls and isoaspartyls. The physiological significance of isoaspartyl formation is emphasized by the presence of the conserved enzyme L-isoaspartyl O methyltransferase (PIMT), whose physiological function appears to be in preventing the accumulation of deamidated proteins. Seemingly consistent with a repair function, overexpression of PIMT in Drosophila melanogaster extends lifespan under conditions expected to contribute to protein damage. Based on structural information and sequence homology we have created mutants of residues proposed to be involved in co-factor binding in Escherichia coli PIMT. Both mutants retain S-adenosyl L-methionine binding capabilities but demonstrate dramatically reduced kinetic capabilities, perhaps suggestive of catalytic roles beyond co-factor binding. As anticipated, overexpression of the wild type enzyme in E. coli results in bacteria with increased tolerance to thermal stress. Surprisingly, even greater levels of heat tolerance were observed with overexpression of the inactive PIMT mutants. The increased survival capabilities observed with overexpression of PIMT in E. coli, and possibly in Drosophila, are not due to increased isoaspartyl repair capabilities but rather a temperature independent induction of the heat shock system as a result of overexpression of a misfolding-prone protein. An alternate hypothesis as to the physiological substrate and function of L-isoaspartyl methyltransferase is proposed. PMID- 14527955 TI - Farnesoid X receptor activates transcription of the phospholipid pump MDR3. AB - The human multidrug resistance gene MDR3 encodes a P-glycoprotein that belongs to the ATP-binding cassette transporter family (ABCB4). MDR3 is a critical trans locator for phospholipids across canalicular membranes of hepatocytes, evidenced by the fact that human MDR3 deficiencies result in progressive familial intrahepatic cholestasis type III. It has been reported previously that MDR3 expression is modulated by hormones, cellular stress, and xenobiotics. Here we show that the MDR3 gene is trans-activated by the farnesoid X receptor (FXR) via a direct binding of FXR/retinoid X receptor alpha heterodimers to a highly conserved inverted repeat element (a FXR response element) at the distal promoter (-1970 to -1958). In FXR trans-activation assays, both the endogenous FXR agonist chenodeoxycholate and the synthetic agonist GW4064 activated the MDR3 promoter. Deletion or mutation of this inverted repeat element abolished FXR-mediated MDR3 promoter activation. Consistent with these data, MDR3 mRNA was significantly induced by both chenodeoxycholate and GW4064 in primary human hepatocytes in time and dose-dependent fashions. In conclusion, we demonstrate that MDR3 expression is directly up-regulated by FXR. These results, together with the previous report that the bile salt export pump is a direct FXR target, suggest that FXR coordinately controls secretion of bile salts and phospholipids. Results of this study further support the notion that FXR is a master regulator of lipid metabolism. PMID- 14527956 TI - The human phosphatidylinositol phosphatase SAC1 interacts with the coatomer I complex. AB - The Saccharomyces cerevisiae SAC1 gene encodes an integral membrane protein of the endoplasmic reticulum (ER) and the Golgi apparatus. Yeast SAC1 mutants display a wide array of phenotypes including inositol auxotrophy, cold sensitivity, secretory defects, disturbed ATP transport into the ER, or suppression of actin gene mutations. At present, it is not clear how these phenotypes relate to the finding that SAC1 displays polyphosphoinositide phosphatase activity. Moreover, it is still an open question whether SAC1 functions similarly in mammalian cells, since some phenotypes are yeast-specific. Potential protein interaction partners and, connected to that, possible regulatory circuits have not been described. Therefore, we have cloned human SAC1 (hSAC1), show that it behaves similar to ySac1p in terms of substrate specificity, demonstrate that the endogenous protein localizes to the ER and Golgi, and identify for the first time members of the coatomer I (COPI) complex as interaction partners of hSAC1. Mutation of a putative COPI interaction motif (KXKXX) at its C terminus abolishes interaction with COPI and causes accumulation of hSAC1 in the Golgi. In addition, we generated a catalytically inactive mutant, demonstrate that its lipid binding capacity is unaltered, and show that it accumulates in the Golgi, incapable of interacting with the COPI complex despite the presence of the KXKXX motif. These results open the possibility that the enzymatic function of hSAC1 provides a switch for accessibility of the COPI interaction motif. PMID- 14527957 TI - Efficient targeting of conserved cryptic epitopes of infectious agents by single domain antibodies. African trypanosomes as paradigm. AB - Antigen variation is a successful defense system adopted by several infectious agents to evade the host immune response. The principle of this defense strategy in the African trypanosome paradigm involves a dense packing of variant surface glycoproteins (VSG) exposing only highly variable and immuno-dominant epitopes to the immune system, whereas conserved epitopes become inaccessible for large molecules. Reducing the size of binders that target the conserved, less immunogenic, cryptic VSG epitopes forms an obvious solution to combat these parasites. This goal was achieved by introducing dromedary Heavy-chain antibodies. We found that only these unique antibodies recognize epitopes common to multiple VSG classes. After phage display of their antigen-binding repertoire, we isolated a single domain antibody fragment with high specificity for the conserved Asn-linked carbohydrate of VSG. In sharp contrast to labeled concanavalin-A that stains only the flagellar pocket where carbohydrates are accessible because of less dense VSG packing, the single domain binder stains the entire surface of viable parasites, irrespective of the VSG type expressed. This corroborates the idea that small antibody fragments, but not larger lectins or conventional antibody fragments, are able to penetrate the dense VSG coat to target their epitope. The diagnostic potential of this fluorescently labeled binder was proven by the direct, selective, and sensitive detection of parasites in blood smears. The employment of this binder as a molecular recognition unit in immuno-toxins designed for trypanosomosis therapy becomes feasible as well. This was illustrated by the specific trypanolysis induced by an antibody::beta lactamase fusion activating a prodrug. PMID- 14527958 TI - Transcriptional activation of mouse mast cell Protease-7 by activin and transforming growth factor-beta is inhibited by microphthalmia-associated transcription factor. AB - Previous studies have revealed that activin A and transforming growth factor beta1 (TGF-beta1) induced migration and morphological changes toward differentiation in bone marrow-derived cultured mast cell progenitors (BMCMCs). Here we show up-regulation of mouse mast cell protease-7 (mMCP-7), which is expressed in differentiated mast cells, by activin A and TGF-beta1 in BMCMCs, and the molecular mechanism of the gene induction of mmcp-7. Smad3, a signal mediator of the activin/TGF-beta pathway, transcriptionally activated mmcp-7. Microphthalmia-associated transcription factor (MITF), a tissue-specific transcription factor predominantly expressed in mast cells, melanocytes, and heart and skeletal muscle, inhibited Smad3-mediated mmcp-7 transcription. MITF associated with Smad3, and the C terminus of MITF and the MH1 and linker region of Smad3 were required for this association. Complex formation between Smad3 and MITF was neither necessary nor sufficient for the inhibition of Smad3 signaling by MITF. MITF inhibited the transcriptional activation induced by the MH2 domain of Smad3. In addition, MITF-truncated N-terminal amino acids could associate with Smad3 but did not inhibit Smad3-mediated transcription. The level of Smad3 was decreased by co-expression of MITF but not of dominant-negative MITF, which resulted from proteasomal protein degradation. The changes in the level of Smad3 protein were paralleled by those in Smad3-mediated signaling activity. These findings suggest that MITF negatively regulates Smad-dependent activin/TGF-beta signaling in a tissue-specific manner. PMID- 14527959 TI - Induction of cIAP-2 in human colon cancer cells through PKC delta/NF-kappa B. AB - Activation of protein kinase C (PKC) prevents apoptosis in certain cells; however, the mechanisms are largely unknown. Inhibitors of apoptosis (IAP) family members, including NAIP, cIAP-1, cIAP-2, XIAP/hILP, survivin, and BRUCE, block apoptosis by binding and potently inhibiting caspases. Activation of NF-kappa B contributes to cIAP-2 induction; however, the cellular mechanisms regulating cIAP 2 expression have not been entirely defined. In this study, we examined the role of the PKC and NF-kappa B pathways in the regulation of cIAP-2 in human colon cancers. We found that cIAP-2 mRNA levels were markedly increased in human colon cancer cells by treatment with the phorbol ester, phorbol-12-myristate-13-acetate (PMA), or bryostatin 1. Inhibitors of the Ca2+-independent, novel PKC isoforms, but not inhibitors of MAPK, PI3-kinase, or PKA, blocked PMA-stimulated cIAP-2 mRNA expression, suggesting a role of PKC in PMA-mediated cIAP-2 induction. Pretreatment with the PKC delta-selective inhibitor rottlerin or transfection with an antisense PKC delta oligonucleotide inhibited PMA-induced cIAP-2 expression, whereas cotransfection with a PKC delta plasmid induced cIAP-2 promoter activity, which, taken together, identifies a role for PKC delta in cIAP 2 induction. Treatment with the proteasome inhibitor, MG132 or inhibitors of NF kappa B (e.g. PDTC and gliotoxin), decreased PMA-induced up-regulation of cIAP-2. PMA-induced NF-kappa B activation was blocked by either GF109203x, MG132, PDTC, or gliotoxin. Moreover, overexpression of PKC delta-induced cIAP-2 promoter activity and increased NF-kappa B transactivation, suggesting regulation of cIAP 2 expression by a PKC delta/NF-kappa B pathway. In conclusion, our findings demonstrate a role for a PKC/NF-kappa B-dependent pathway in the regulation of cIAP-2 expression in human colon cancer cells. These data suggest a novel mechanism for the anti-apoptotic function mediated by the PKC delta/NF-kappa B/cIAP-2 pathway in certain cancers. PMID- 14527960 TI - cPKC-dependent sequestration of membrane-recycling components in a subset of recycling endosomes. AB - In addition to the classical role of protein kinase C (PKC) as a mediator of transmembrane signals initiated at the plasma membrane, there is also significant evidence to suggest that a more sustained PKC activity is necessary for a variety of long term cellular responses. To date, the subcellular localization of PKC during sustained activation has not been extensively studied. We report here that long term activation of PKC (1 h) leads to the selective translocation of classical PKC isoenzymes, alpha and betaII, to a juxtanuclear compartment. Juxtanuclear translocation of PKC required an intact C1 and C2 domain, and occurred in a microtubule-dependent manner. This juxtanuclear compartment was localized close to the Golgi complex but displayed no overlap with Golgi markers, and was resistant to dispersal with Golgi disrupting agents, brefeldin A and nocodazole. Further characterization revealed that PKCalpha and betaII translocated to a compartment that colocalized with the small GTPase, rab11, which is a marker for the subset of recycling endosomes concentrated around the microtubule-organizing center/centrosome. Analysis of the functional consequence of cPKC translocation on membrane recycling demonstrated a cPKC-dependent sequestration of transferrin, a marker of membrane recycling, in the cPKC compartment. These results identify a novel site for cPKC translocation and define a novel function for the sustained activation of PKCalpha and betaII in regulation of recycling components. PMID- 14527961 TI - C3b2-IgG complexes retain dimeric C3 fragments at all levels of inactivation. AB - C3b2-IgG complexes are formed during complement activation in serum by attachment of two C3b molecules (the proteolytically activated form of C3) to one IgG heavy chain (IgG HC) via ester bonds. Because of the presence of two C3b molecules, these complexes are very efficient activators of the alternative complement pathway. Likewise, dimeric C3b is known to enhance complement receptor 1 dependent phagocytosis, and dimeric C3d (the smallest thioester-containing fragment of C3) linked to a protein antigen facilitates CR2-dependent B-cell proliferation. Because the efficiency of all these interactions depends on the number of C3 fragments, we investigated whether C3b2-IgG complexes retained dimeric structure upon physiological inactivation. We used two-dimensional SDS PAGE and Western blot to study the arrangement of the C3b molecules by analyzing the fragmentation pattern after cleavage of the ester bonds. Upon inactivation with factors H and I, a 185-kDa band was generated under reducing conditions. It released IgG HC and the 65-kDa fragment of C3b alpha' chain after hydrolysis of the ester bonds with hydroxylamine. The two C3b molecules were not 65-kDa-to-40 kDa linked, because neither ester-bonded 65 kDa HC nor 65 kDa-40 kDa fragments were observed, nor was a 40-kDa peptide released after hydroxylamine cleavage. Factor I and CR1 cleaved the C3b2-IgG molecule to its final physiological product, C3dg2-IgG, which migrated as a 133-kDa fragment in reduced form. This fragment released exclusively C3dg (the final physiological product of C3b inactivation by factor I) and IgG HC. C3dg2-HC appeared as a double band on SDS PAGE only at low gel porosity, suggesting the presence of two conformers of the same composition. Our results suggest that, upon physiological inactivation, C3b2 IgG complexes retain dimeric inactivated C3b and C3dg, which allows bivalent binding to the corresponding complement receptors. PMID- 14527962 TI - Assessment of skeletal muscle mass in men with spinal cord injury using dual energy X-ray absorptiometry and magnetic resonance imaging. AB - The purpose of this study was to determine whether the proportion of skeletal muscle in the fat-free soft tissue mass (FFST) is the same in men with spinal cord injury (SCI) and able-bodied controls. Skeletal muscle mass and FFST of the midthigh were determined by using magnetic resonance imaging and dual-energy X ray absorptiometry, respectively, in men with long-term (>2 yr) complete SCI (n = 8) and able-bodied controls of similar age, height, and weight (n = 8). Muscle mass (1.36 +/- 0.77 vs. 2.44 +/- 0.47 kg) and FFST (1.70 +/- 0.94 vs. 2.73 +/- 0.80 kg) were lower in the SCI group than in the controls (P < 0.05), but the lower ratio of muscle to FFST in the SCI group (0.80 +/- 0.09 vs. 0.91 +/- 0.10, P < 0.05) suggested that they had a lower proportion of muscle in the FFST than in controls. This notion was supported by analysis of covariance, in that the mean muscle adjusted to the mean FFST of the groups combined was lower in the SCI group. Despite the lower proportion of muscle in the FFST of the SCI group, the relation between muscle and FFST was strong in the SCI group (r = 0.99) and controls (r = 0.96). The findings suggest a disproportionate loss of muscle in the paralyzed thighs after SCI relative to other nonfat constituents, which may be accurately estimated in men with long-term SCI by dual-energy X-ray absorptiometry if the lower proportion of muscle in the FFST (approximately 15%) is taken into account. PMID- 14527963 TI - Reduction in heat-induced gastrointestinal hyperpermeability in rats by bovine colostrum and goat milk powders. AB - Male Sprague-Dawley rats were assigned to one of three dietary groups [standard diet (Cont; n = 8), standard diet plus bovine colostrum powder (BColost 1.7 g/kg; n = 8), or goat milk powder (GMilk 1.7 g/kg; n = 8)] to determine the ability of these supplements to reduce gastrointestinal hyperpermeability induced by heat. Raising core body temperature of rats to 41.5 degrees C increased transfer of (51)Cr-EDTA from gut into blood 34-fold relative to the ambient temperature value (P < 0.05) in the Cont group of rats, indicative of increased gastrointestinal permeability. Significantly less (P < 0.01) (51)Cr-EDTA was transferred into the blood of rats in either the BColost (27% of Cont) or GMilk group (10% of Cont) after heating, showing that prior supplementation with either bovine colostrum or goat milk powder significantly reduced the impact of heat stress on gastrointestinal permeability. The changes in the BColost group were not significantly different than those of the GMilk group. The potential mechanism of the protective effect of bovine colostrum and goat milk powders may involve modulation of tight junction permeability, because both powders were able to maintain transepithelial resistance in Madin Darby canine kidney cells challenged with EGTA compared with cells maintained in media only. The results show that bovine colostrum powder can partially alleviate the effects of hyperthermia on gastrointestinal permeability in the intact animal. Moreover, goat milk powder was equally as effective as bovine colostrum powder, and both may be of benefit in other situations where gastrointestinal barrier function is compromised. PMID- 14527965 TI - On the reflex coactivation of ankle flexor and extensor muscles induced by a sudden drop of support surface during walking in humans. AB - Recent studies have revealed that the stretch reflex responses of both ankle flexor and extensor muscles are coaugmented in the early stance phase of human walking, suggesting that these coaugmented reflex responses contribute to secure foot stabilization around the heel strike. To test whether the reflex responses mediated by the stretch reflex pathway are actually induced in both the ankle flexor and extensor muscles when the supportive surface is suddenly destabilized, we investigated the electromyographic (EMG) responses induced after a sudden drop of the supportive surface at the early stance phase of human walking. While subjects walked on a walkway, the specially designed movable supportive surface was unexpectedly dropped 10 mm during the early stance phase. The results showed that short-latency reflex EMG responses after the impact of the drop (<50 ms) were consistently observed in both the ankle flexor and extensor muscles in the perturbed leg. Of particular interest was that a distinct response appeared in the tibialis anterior muscle, although this muscle showed little background EMG activity during the stance phase. These results indicated that the reflex activities in the ankle muscles certainly acted when the supportive surface was unexpectedly destabilized just after the heel strike during walking. These reflex responses were most probably mediated by the facilitated stretch reflex pathways of the ankle muscles at the early stance phase and were suggested to be relevant to secure stabilization around the ankle joint during human walking. PMID- 14527964 TI - Blockade of the sympathetic nervous system degrades ligament in a rat MCL model. AB - We hypothesize that blockade of the sympathetic nervous system degrades ligament. We tested this hypothesis in a rat medial collateral ligament (MCL) model. Fifteen animals were treated for 10 days with the sympathetic chemotoxin guanethidine using osmotic pumps, whereas 15 control rats received pumps containing saline. A reduction in plasma concentrations of norepinephrine in the guanethidine rats indicated a significant decrease in sympathetic nerve activity. Vasoactive intestinal peptide and neuropeptide Y were decreased in MCLs from guanethidine animals, as quantified by radioimmunoassays. Tissue vascularity was substantially increased in guanethidine MCLs, whereas mechanical properties were significantly decreased. Proteases, such as matrix metalloproteinases (MMP) and cysteine proteases, play a major role in ligament degradation. The proteases MMP 13, cathepsin K, and tartrate-resistant acid phosphatase (TRAP) have collagenolytic activity and have been shown in rat ligament tissues. To determine whether the degradation seen in this study was due to protease activity, we determined the expression of these enzymes in control and treated MCLs. Real-time quantitative PCR revealed that guanethidine treatment increased expression of MMP 13 and cathepsin K mRNAs, although overall expression levels of MMP-13 and TRAP were relatively low. Histology also identified increases in TRAP and cathepsin K, but not MMP-13, in guanethidine-treated tissues. Results support our hypothesis that blockade of the sympathetic nervous system substantially degrades ligament. PMID- 14527966 TI - Regulation of the angiotensin-converting enzyme activity by a time-course hypoxia in the carotid body. AB - Chronic hypoxia activates a local angiotensin-generating system in the carotid body. Here, we test the hypothesis that the activity of the critical enzyme for this system, angiotensin-converting enzyme (ACE), in the carotid body is subject to regulation by a time-course hypoxia. Results from the carotid body assays showed that ACE activity was markedly increased under the hypoxic stress of 7-, 14-, 21-, and 28-day exposures. The changes in ACE activity of 7-day (15.00 vs. 30.95 x 10(-5) nmol.microg(-1).min(-1)), 14-day (8.73 vs. 30.25 x 10(-5) nmol.microg(-1).min(-1)), and 21-day (11.41 vs. 31.83 x 10(-5) nmol.microg( 1).min(-1)) hypoxia treatments were enhanced significantly. However, ACE activity in 28-day (13.18 vs. 24.53 x 10(-5) nmol.microg(-1).min(-1)) hypoxia treatment was observed to increase insignificantly when compared with results in the respective control groups. Captopril inhibited all rises in ACE activity in both the control and experimental groups. Results clearly indicate an activation of the enzymatic activity of ACE, the critical enzyme for determining the conversion of angiotensin I into the physiologically active angiotensin II, by chronic hypoxia in the carotid body. An increase in the ACE activity may increase the local production of angiotensin II in the carotid body and thus its agonist action at the AT1 receptor. This may be important in the modulation of cardiopulmonary adaptation in the hypoxic ventilatory response as well as for electrolyte and water homeostasis during chronic hypoxia. PMID- 14527967 TI - Coordination of intrinsic and extrinsic tongue muscles during spontaneous breathing in the rat. AB - The muscular-hydrostat model of tongue function proposes a constant interaction of extrinsic (external bony attachment, insertion into base of tongue) and intrinsic (origin and insertion within the tongue) tongue muscles in all tongue movements (Kier WM and Smith KK. Zool J Linn Soc 83: 207-324, 1985). Yet, research that examines the respiratory-related effects of tongue function in mammals continues to focus almost exclusively on the respiratory control and function of the extrinsic tongue protrusor muscle, the genioglossus muscle. The respiratory control and function of the intrinsic tongue muscles are unknown. Our purpose was to determine whether intrinsic tongue muscles have a respiration related activity pattern and whether intrinsic tongue muscles are coactivated with extrinsic tongue muscles in response to respiratory-related sensory stimuli. Esophageal pressure and electromyographic (EMG) activity of an extrinsic tongue muscle (hyoglossus), an intrinsic tongue muscle (superior longitudinal), and an external intercostal muscle were studied in anesthetized, tracheotomized, spontaneously breathing rats. Mean inspiratory EMG activity was compared at five levels of inspired CO2. Intrinsic tongue muscles were often quiescent during eupnea but active during hypercapnia, whereas extrinsic tongue muscles were active in both eupnea and hypercapnia. During hypercapnia, the activities of the airway muscles were largely coincident, although the onset of extrinsic muscle activity generally preceded the onset of intrinsic muscle activation. Our findings provide evidence, in an in vivo rodent preparation, of respiratory modulation of motoneurons supplying intrinsic tongue muscles. Distinctions noted between intrinsic and extrinsic activities could be due to differences in motoneuron properties or the central, respiration-related control of each motoneuron population. PMID- 14527969 TI - Influence of tendon slack on electromechanical delay in the human medial gastrocnemius in vivo. AB - The purpose of this study was to clarify the influence of muscle-tendon complex stretch on electromechanical delay (EMD) in terms of the extent of tendon slack in the human medial gastrocnemius (MG). EMD and MG tendon length were measured at each of five ankle joint angles (-30, -20, -10, 0, and 5 degrees : positive values for dorsiflexion) using percutaneous electrical stimulation and ultrasonography, respectively. The extent of MG tendon slack was calculated as MG tendon length shortening, standardized with MG tendon slack length obtained at the joint angle (-16 degrees +/- 5 degrees ) where the passive ankle joint torque was zero. EMD at -30 degrees (19.2 +/-2.2 ms) and -20 degrees (17.2 +/- 1.3 ms) was significantly greater than that at -10 degrees (16.0 +/-2.3 ms), 0 degrees (15.0 +/-1.4 ms), and 5 degrees (14.8 +/-1.4 ms), and at 0 and 5 degrees, respectively. The relative EMD, normalized with the maximal EMD for each subject, decreased dependent on the extent of decrease in MG tendon slack. There were no significant differences in EMD among the joint angles (-10, 0, and 5 degrees ) where MG tendon slack was taken up. These results suggest that the extent of tendon slack is an important factor for determining EMD. PMID- 14527968 TI - Development of REM sleep drive and clinical implications. AB - Rapid eye movement (REM) sleep in the human declines from approximately 50% of total sleep time ( approximately 8 h) in the newborn to approximately 15% of total sleep time (approximately 1 h) in the adult, and this decrease takes place mainly between birth and the end of puberty. We hypothesize that without this developmental decrease in REM sleep drive, lifelong increases in REM sleep drive may ensue. In the rat, the developmental decrease in REM sleep occurs 10-30 days after birth, declining from >70% of total sleep time in the newborn to the adult level of approximately 15% of sleep time during this period. Rats at 12-21 days of age were anesthetized with ketamine and decapitated, and brain stem slices were cut for intracellular recordings. We found that excitatory responses of pedunculopontine nucleus (PPN) neurons to N-methyl-D-aspartic acid decrease, while responses to kainic acid increase, over this critical period. During this developmental period, inhibitory responses to serotonergic type 1 agonists increase but responses to serotonergic type 2 agonists do not change. The results suggest that as PPN neurons develop, they are increasingly activated by kainic acid and increasingly inhibited by serotonergic type 1 receptors. These processes may be related to the developmental decrease in REM sleep. Developmental disturbances in each of these systems could induce differential increases in REM sleep drive, accounting for the postpubertal onset of a number of different disorders manifesting increases in REM sleep drive. Examination of modulation by PPN projections to ascending and descending targets revealed the presence of common signals modulating ascending arousal-related functions and descending postural/locomotor-related functions. PMID- 14527970 TI - Comparative impact of guidelines, clinical data, and decision support on prescribing decisions: an interactive web experiment with simulated cases. AB - OBJECTIVE: The aim of this study was to compare the clinical impact of computerized decision support with and without electronic access to clinical guidelines and laboratory data on antibiotic prescribing decisions. DESIGN: A crossover trial was conducted of four levels of computerized decision support-no support, antibiotic guidelines, laboratory reports, and laboratory reports plus a decision support system (DSS), randomly allocated to eight simulated clinical cases accessed by the Web. MEASUREMENTS: Rate of intervention adoption was measured by frequency of accessing information support, cost of use was measured by time taken to complete each case, and effectiveness of decision was measured by correctness of and self-reported confidence in individual prescribing decisions. Clinical impact score was measured by adoption rate and decision effectiveness. RESULTS: Thirty-one intensive care and infectious disease specialist physicians (ICPs and IDPs) participated in the study. Ventilator associated pneumonia treatment guidelines were used in 24 (39%) of the 62 case scenarios for which they were available, microbiology reports in 36 (58%), and the DSS in 37 (60%). The use of all forms of information support did not affect clinicians' confidence in their decisions. Their use of the DSS plus microbiology report improved the agreement of decisions with those of an expert panel from 65% to 97% (p=0.0002), or to 67% (p=0.002) when antibiotic guidelines only were accessed. Significantly fewer IDPs than ICPs accessed information support in making treatment decisions. On average, it took 245 seconds to make a decision using the DSS compared with 113 seconds for unaided prescribing (p<0.001). The DSS plus microbiology reports had the highest clinical impact score (0.58), greater than that of electronic guidelines (0.26) and electronic laboratory reports (0.45). CONCLUSION: When used, computer-based decision support significantly improved decision quality. In measuring the impact of decision support systems, both their effectiveness in improving decisions and their likely rate of adoption in the clinical environment need to be considered. Clinicians chose to use antibiotic guidelines for one third and microbiology reports or the DSS for about two thirds of cases when they were available to assist their prescribing decisions. PMID- 14527971 TI - A methodology and implementation for annotating digital images for context appropriate use in an academic health care environment. AB - Use of digital medical images has become common over the last several years, coincident with the release of inexpensive, mega-pixel quality digital cameras and the transition to digital radiology operation by hospitals. One problem that clinicians, medical educators, and basic scientists encounter when handling images is the difficulty of using business and graphic arts commercial-off-the shelf (COTS) software in multicontext authoring and interactive teaching environments. The authors investigated and developed software-supported methodologies to help clinicians, medical educators, and basic scientists become more efficient and effective in their digital imaging environments. The software that the authors developed provides the ability to annotate images based on a multispecialty methodology for annotation and visual knowledge representation. This annotation methodology is designed by consensus, with contributions from the authors and physicians, medical educators, and basic scientists in the Departments of Radiology, Neurobiology and Anatomy, Dermatology, and Ophthalmology at the University of Utah. The annotation methodology functions as a foundation for creating, using, reusing, and extending dynamic annotations in a context-appropriate, interactive digital environment. The annotation methodology supports the authoring process as well as output and presentation mechanisms. The annotation methodology is the foundation for a Windows implementation that allows annotated elements to be represented as structured eXtensible Markup Language and stored separate from the image(s). PMID- 14527972 TI - Development of a Web-based event reporting system in an academic environment. AB - In pursuit of a strategy for patient safety and error reduction, The Ohio State University Health System developed and implemented a standardized voluntary event reporting system. The Web-based application is user friendly as well as context sensitive and encompasses a broad range of errors, events, and near misses. A full organizational transformation was required to effectively implement the system, which involved process reengineering for event entry and for postentry automated workflows. This system serves as the foundation for efficient and consistent reporting processes, which are essential for encouraging a culture of commitment to patient safety. PMID- 14527973 TI - Data quality of general practice electronic health records: the impact of a program of assessments, feedback, and training. AB - OBJECTIVE: The aim of this study was to investigate the impact of a program of repeated assessments, feedback, and training on the quality of coded clinical data in general practice. DESIGN: A prospective uncontrolled intervention study was conducted in a general practice research network. MEASUREMENTS: Percentage of recorded consultations with a coded problem title and percentage of patients receiving a specific drug (e.g., tamoxifen) who had the relevant morbidity code (e.g., breast cancer) were calculated. Annual period prevalence of 12 selected morbidities was compared with parallel data derived from the fourth National Study of Morbidity Statistics from General Practice (MSGP4). RESULTS: The first two measures showed variation between practices at baseline, but on repeat assessments all practices improved or maintained their levels of coding. The period prevalence figures also were variable, but over time rates increased to levels comparable with, or above, MSGP4 rates. Practices were able to provide time and resources for feedback and training sessions. CONCLUSION: A program of repeated assessments, feedback, and training appears to improve data quality in a range of practices. The program is likely to be generalizable to other practices but needs a trained support team to implement it that has implications for cost and resources. PMID- 14527974 TI - Human factors barriers to the effective use of ten HIV clinical reminders. AB - OBJECTIVE: Substantial variations in adherence to guidelines for human immunodeficiency virus (HIV) care have been documented. To evaluate their effectiveness in improving quality of care, ten computerized clinical reminders (CRs) were implemented at two pilot and eight study sites. The aim of this study was to identify human factors barriers to the use of these CRs. DESIGN: Observational study was conducted of CRs in use at eight outpatient clinics for one day each and semistructured interviews were conducted with physicians, pharmacists, nurses, and case managers. MEASUREMENTS: Detailed handwritten field notes of interpretations and actions using the CRs and responses to interview questions were used for measurement. RESULTS: Barriers present at more than one site were (1) workload during patient visits (8 of 8 sites), (2) time to document when a CR was not clinically relevant (8 of 8 sites), (3) inapplicability of the CR due to context-specific reasons (9 of 26 patients), (4) limited training on how to use the CR software for rotating staff (5 of 8 sites) and permanent staff (3 of 8 sites), (5) perceived reduction of quality of provider-patient interaction (3 of 23 permanent staff), and (6) the decision to use paper forms to enable review of resident physician orders prior to order entry (2 of 8 sites). CONCLUSION: Six human factors barriers to the use of HIV CRs were identified. Reducing these barriers has the potential to increase use of the CRs and thereby improve the quality of HIV care. PMID- 14527975 TI - A conceptual framework for evaluating outpatient electronic prescribing systems based on their functional capabilities. AB - OBJECTIVE: Electronic prescribing (e-prescribing) may substantially improve health care quality and efficiency, but the available systems are complex and their heterogeneity makes comparing and evaluating them a challenge. The authors aimed to develop a conceptual framework for anticipating the effects of alternative designs for outpatient e-prescribing systems. DESIGN: Based on a literature review and on telephone interviews with e-prescribing vendors, the authors identified distinct e-prescribing functional capabilities and developed a conceptual framework for evaluating e-prescribing systems' potential effects based on their capabilities. Analyses of two commercial e-prescribing systems are presented as examples of applying the conceptual framework. MEASUREMENTS: Major e prescribing functional capabilities identified and the availability of evidence to support their specific effects. RESULTS: The proposed framework for evaluating e-prescribing systems is organized using a process model of medication management. Fourteen e-prescribing functional capabilities are identified within the model. Evidence is identified to support eight specific effects for six of the functional capabilities. The evidence also shows that a functional capability with generally positive effects can be implemented in a way that creates unintended hazards. Applying the framework involves identifying an e-prescribing system's functional capabilities within the process model and then assessing the effects that could be expected from each capability in the proposed clinical environment. CONCLUSION: The proposed conceptual framework supports the integration of available evidence in considering the full range of effects from e prescribing design alternatives. More research is needed into the effects of specific e-prescribing functional alternatives. Until more is known, e prescribing initiatives should include provisions to monitor for unintended hazards. PMID- 14527976 TI - PalmCIS: a wireless handheld application for satisfying clinician information needs. AB - Wireless handheld technology provides new ways to deliver and present information. As with any technology, its unique features must be taken into consideration and its applications designed accordingly. In the clinical setting, availability of needed information can be crucial during the decision-making process. Preliminary studies performed at New York Presbyterian Hospital (NYPH) determined that there are inadequate access to information and ineffective communication among clinicians (potential proximal causes of medical errors). In response to these findings, the authors have been developing extensions to their Web-based clinical information system including PalmCIS, an application that provides access to needed patient information via a wireless personal digital assistant (PDA). The focus was on achieving end-to-end security and developing a highly usable system. This report discusses the motivation behind PalmCIS, design and development of the system, and future directions. PMID- 14527977 TI - The InterMed approach to sharable computer-interpretable guidelines: a review. AB - InterMed is a collaboration among research groups from Stanford, Harvard, and Columbia Universities. The primary goal of InterMed has been to develop a sharable language that could serve as a standard for modeling computer interpretable guidelines (CIGs). This language, called GuideLine Interchange Format (GLIF), has been developed in a collaborative manner and in an open process that has welcomed input from the larger community. The goals and experiences of the InterMed project and lessons that the authors have learned may contribute to the work of other researchers who are developing medical knowledge based tools. The lessons described include (1) a work process for multi institutional research and development that considers different viewpoints, (2) an evolutionary lifecycle process for developing medical knowledge representation formats, (3) the role of cognitive methodology to evaluate and assist in the evolutionary development process, (4) development of an architecture and (5) design principles for sharable medical knowledge representation formats, and (6) a process for standardization of a CIG modeling language. PMID- 14527980 TI - Challenges and opportunities for primary care evaluation. PMID- 14527978 TI - Take note(s): differential EHR satisfaction with two implementations under one roof. AB - OBJECTIVE: The aim of this study was to rigorously evaluate perceived differences in satisfaction with an electronic health record (EHR) between residents of two medical specialties who share the same health record, practice location, administration, and information technology support. DESIGN: A cross-sectional survey was used comparing user satisfaction between pediatrics residents and internal medicine residents in an academic practice. MEASUREMENTS: The survey was designed to measure baseline demographic characteristics, attitudes toward computers, general satisfaction with an EHR, and perceived practicality of use, variation from familiar practice, organizational support, and impact on delivery of care. RESULTS: Medicine subjects were similar to pediatrics subjects in baseline demographic characteristics. Satisfaction with the EHR implementation was very high for both sets of subjects, but internal medicine residents were significantly less likely to be satisfied with the EHR implementation (relative risk [RR]=0.84, 95% confidence interval [CI]=0.73-0.98) and considerably less likely to believe that their colleagues were satisfied with it (RR=0.56, 95% CI=0.41-0.77). The only surveyed characteristic independently predicting satisfaction was medical specialty (p=0.04). Medicine subjects were less likely to believe template-based documentation improved their efficiency (RR=0.64, 95% CI=0.46-0.88). They were significantly more likely to believe the system had been designed to improve billing (RR=1.50, 95% CI=1.05-2.04) and not to improve patient care (RR=0.61, 95% CI=0.44-0.85). CONCLUSION: The authors found a difference in satisfaction between internal medicine and pediatrics users of an EHR. Although many potential factors that influence satisfaction were similar between subjects in the two specialties, differences in previous experience may have influenced the results. Medicine residents had more previous experience with a different EHR implementation, which they may have perceived as superior to the one involved in this study. Pediatric residents had more previous experience with structured data entry prior to EHR implementation and more preventive care patient encounters for which structured data entry may be well suited. Since successful implementations generally require satisfied users, understanding what factors affect satisfaction can improve chances of a system's success. PMID- 14527981 TI - Improving professional competence--the way ahead? AB - Globally, there has been considerable interest from the public, medical profession, and governments in ensuring that doctors are fit for practice. Appropriate accountability for the profession has become an important contemporary issue. With doubts emerging on the effectiveness of professionally led regulation, a variety of different mechanisms have been devised to ensure that patients receive appropriate and good quality care. This paper reviews regulation of the medical profession and indicates the way in which doctors in the UK are held accountable for their practice. PMID- 14527982 TI - Conceptual frameworks for health systems performance: a quest for effectiveness, quality, and improvement. AB - ISSUES: Countries and international organizations have recently renewed their interest in how health systems perform. This has led to the development of performance indicators for monitoring, assessing, and managing health systems to achieve effectiveness, equity, efficiency, and quality. Although the indicators populate conceptual frameworks, it is often not very clear just what the underlying concepts might be or how effectiveness is conceptualized and measured. Furthermore, there is a gap in the knowledge of how the resultant performance data are used to stimulate improvement and to ensure health care quality. ADDRESSING THE ISSUES: This paper therefore explores, individually, the conceptual bases, effectiveness and its indicators, as well as the quality improvement dynamics of the performance frameworks of the UK, Canada, Australia, US, World Health Organization, and Organisation for Economic Co-operation and Development. RESULTS: We see that they all conceive health and health system performance in one or more supportive frameworks, but differ in concepts and operations. Effectiveness often implies, nationally, the achievement of high quality outcomes of care, or internationally, the efficient achievement of system objectives, or both. Its indicators are therefore mainly outcome and, less so, process measures. The frameworks are linked to a combination of tools and initiatives to stimulate and manage performance and quality improvement. CONCLUSIONS: These dynamics may ensure the proper environment for these conceptual frameworks where, alongside objectives such as equity and efficiency, effectiveness (therefore, quality) becomes the core of health systems performance. PMID- 14527983 TI - Predictors of variation in office visit interval assignment. AB - OBJECTIVE: Despite the important influence of ambulatory appointment revisit intervals (RVI) on access to care, physicians receive no formal training in this area and research indicates that there is significant practice variation. Our objective was to examine whether predictors of RVI assignment that we had assessed using vignettes were also significant in the actual patient care setting. DATA SOURCES AND STUDY DESIGN: A cross-sectional survey of 59 internal medicine residents collected at the end of office visits for patients with hypertension or diabetes. Two hundred and twenty-eight patients seen in 1997 for continuity care in two academic clinics in New Orleans, Louisiana. DATA COLLECTION: The main outcome was RVI in weeks. We assessed the relationship between physician, visit, and patient-level covariates, and RVI assignment in univariate and multivariate analyses using hierarchical linear models. PRINCIPAL FINDINGS: The mean RVI was 12.4 weeks (range 1-42 weeks) and was similar for patients with diabetes and hypertension. The final model accounted for 35.7% of the variance in RVI assignment and included: perceptions of the patient's systolic blood pressure, disease stability, and compliance; comorbidity, physician age, sex, and identity; and changing therapy for the primary diagnosis. The identity of the physician was the largest contributor to the variance, accounting for 14.7%. CONCLUSIONS: Intrinsic characteristics of physicians and their subjective interpretations of their patients' disease stability are the most important determinants of ambulatory RVI assignment. Intervening to reduce this variation in practice is challenging because limited research is currently available on the optimum RVI for patients with chronic illnesses such as diabetes and hypertension. PMID- 14527984 TI - A reliability study of an instrument for measuring general practitioner consultation skills: the LIV-MAAS scale. AB - OBJECTIVE: To evaluate the reliability of a new tool, the LIV-MAAS, in assessing consultation competence in UK general practice. DESIGN: These were pilot studies, with small numbers of participants. Videoed general practitioner (GP) consultations were analysed by trained lay and professional raters, using the LIV MAAS. The inter-rater reliabilities were assessed. Four videos were assessed by five raters in a pilot study. After this, 71 consultations from eight doctors were assessed by sets of three raters. MAIN MEASURES: Inter-rater reliabilities and inter-consultation reliabilities. RESULTS: For the pilot study, the estimated inter-rater reliability ranged from 0.69 (one rater) to 0.91 (five raters). For the main study, the estimated inter-rater reliability for the LIV-MAAS checklist using two raters was 0.71, and using three raters it was 0.78. Mean differences in reliability within each series of nine consultations were 0.20 (three raters) and 0.42 (two raters). CONCLUSIONS: As a measure of 'consultation competence', administered by trained raters (medical or lay) to real GP consultations, the LIV MAAS instrument shows adequate reliability and stability but would benefit from considerable shortening. Further development of the LIV-MAAS and testing with larger samples are required. PMID- 14527986 TI - Effects of state surveillance on new post-hospitalization benzodiazepine use. AB - BACKGROUND: Benzodiazepines (BZD) effectively treat anxiety and insomnia accompanying major health events, including hospitalizations. Prescribing regulations to decrease BZD misuse may negatively impact therapeutic uses. OBJECTIVE: To assess the impact of a Triplicate Prescription Program (TPP) on initiation of post-hospitalization BZD prescribing, both overall and among cardiac and cancer patients in the United States. DESIGN: Interrupted time-series of post-hospitalization BZD dispensing events to enrollees in the US Medicaid program in the states of New York (intervention group) and New Jersey (control group), before and after implementation of a TPP. STUDY PARTICIPANTS: Community dwelling Medicaid enrollees in New York State (n = 67 962) and New Jersey (n = 71 701), hospitalized between 1 January 1988 and 30 November 1990. INTERVENTION: The New York State TPP, implemented on 1 January 1989, requires physicians to prescribe BZD on triplicate prescription forms for state surveillance. OUTCOME MEASURES: Rates and duration of new post-hospitalization use of BZD and substitute medications. RESULTS: Overall, a sudden and sustained 63.5% decrease [95% confidence interval (CI) -58.6% to -68.3%] in new post-hospitalization BZD dispensing-from a baseline rate of 44 discharges with BZD dispensing per 1000 discharges per month-followed the TPP in New York State, without discontinuity in the control state. Patients hospitalized for acute ischemic cardiac events experienced a 72.5% reduction (95% CI -55.5% to -89.4%), and cancer patients a 69.4% reduction (95% CI -36.7% to -100.0%). The TPP did not preferentially reduce BZD use lasting >2 months. Increased substitute use did not offset reductions in BZD use. CONCLUSIONS: By decreasing new short-term post-hospitalization BZD use, the New York State TPP also had unintended effects. PMID- 14527985 TI - Readmissions and the quality of care in patients hospitalized with heart failure. AB - OBJECTIVES: Clinical practice guidelines based on the results of randomized clinical trials recommend that patients with heart failure due to left ventricular systolic dysfunction (LVSD) be treated with angiotensin-converting enzyme inhibitors (ACEI) at doses shown to reduce mortality and readmission. This study examined the relationship between ACEI use at discharge and readmission among patients with heart failure due to LVSD. METHODS AND RESULTS: Data were abstracted from the medical records of 2943 randomly selected patients hospitalized for heart failure in 50 hospitals. The outcome of interest was the number of readmissions occurring up to 21 months after discharge. Six-hundred and eleven patients were eligible for analysis. Compared with patients discharged at a recommended ACEI dose, patients not prescribed an ACEI at discharge had an adjusted rate ratio of readmission (RR) of 1.74 [95% confidence interval (CI) 1.22-2.48], while patients prescribed an ACEI at less than a recommended dose had an RR of 1.24 (95% CI 0.91-1.69) (P = 0.005 for the trend). CONCLUSION: Our results show that ACEI use at discharge in patients with LVSD is associated with decreased rate of readmission. These findings suggest that compliance with the ACEI prescribing recommendations listed in clinical practice guidelines for patients with heart failure due to LVSD confers benefit. PMID- 14527987 TI - Patient-reported quality of life before, compared with after a DRG intervention. AB - OBJECTIVE: To evaluate the effects of the introduction of a payment system based on Diagnosis-Related Groups (DRG) on orthopedic patients' perceptions of quality of life after discharge. DESIGN: A prospective longitudinal study performed between 1992 and 1996. SETTING: Two surgical clinics in the County of Gavleborg, Sweden. STUDY PARTICIPANTS: The selection of patients was defined by surgical treatments/diagnoses, and by geographical area. Out of the 40 consecutively recruited patients per year from each hospital, >85% (n = 145 + 140 = 285) completed a questionnaire 1 week post-discharge, and >75% (n = 128 + 127 = 255) did so at a 6-week follow-up. MAIN OUTCOME MEASURE: The Swedish Health-Related Quality of Life Survey (SWED-QUAL) was used to assess quality of life. Straight factoral analyses of variance (ANOVAs) were performed to compare years of assessment in the same hospital/district. RESULTS: The results demonstrated no pattern of significant pre-post differences in patient-reported quality of life. In all years, most quality of life ratings were lower at 1 week compared with 6 weeks after discharge. These differences were probably the consequence of recently undergone surgery. CONCLUSION: The overall conclusion is that changes implemented in health care, including the DRGs, had no demonstrable impact on patient ratings of quality of life. PMID- 14527988 TI - Lack of clear channels of communication in patient transfer between care facilities leads to fragmentation in care. PMID- 14527989 TI - The use of statistical process control methods in monitoring clinical performance. PMID- 14527992 TI - CT and MR colonography: scanning techniques, postprocessing, and emphasis on polyp detection. AB - In the last decade, computed tomographic (CT) and magnetic resonance (MR) colonography, two new cross-sectional techniques for imaging of the colon, emerged. Both techniques show promising initial results in the detection of polyps equal to or greater than 1 cm in diameter in symptomatic patients. Imaging protocols are still mostly under development and prone to change. Both CT and MR colonography generate a large number of source images, which have to be read carefully for filling defects and, if intravenous contrast material is used, enhancing lesions. An important postprocessing technique is multiplanar reformatting, which allows the viewer to see potential lesions in an orientation other than that of the source images. Virtual endoscopy, a volume rendering technique that generates images from within the colon lumen, is used for problem solving. CT and MR colonography have potential advantages over colonoscopy and double-contrast barium enema examination: multiplanar capabilities, detection of enhancing lesions that make the distinction between fecal residue and true lesion possible, and ante- and retrograde virtual colonoscopy. Currently, a number of studies suggest that patients have a preference for CT colonography over colonoscopy. Patients consider bowel cleansing the most uncomfortable part of any colon examination; hence, from the acceptance point of view, fecal tagging techniques are promising. Before CT and MR colonography can be implemented in daily practice, they must show approximately the same accuracy as colonoscopy for polyp detection in both symptomatic and screening patients. PMID- 14527993 TI - Highly specific zinc finger proteins obtained by directed domain shuffling and cell-based selection. AB - Engineered Cys2His2 zinc finger proteins (ZFPs) can mediate regulation of endogenous gene expression in mammalian cells. Ideally, all zinc fingers in an engineered multifinger protein should be optimized concurrently because cooperative and context-dependent contacts can affect DNA recognition. However, the simultaneous selection of key contacts in even three fingers from fully randomized libraries would require the consideration of >10(24) possible combinations. To address this challenge, we have developed a novel strategy that utilizes directed domain shuffling and rapid cell-based selections. Unlike previously described methods, our strategy is amenable to scale-up and does not sacrifice combinatorial diversity. Using this approach, we have successfully isolated multifinger proteins with improved in vitro and in vivo function. Our results demonstrate that both DNA binding affinity and specificity are important for cellular function and also provide a general approach for optimizing multidomain proteins. PMID- 14527994 TI - A genomic perspective on nutrient provisioning by bacterial symbionts of insects. AB - Many animals show intimate interactions with bacterial symbionts that provision hosts with limiting nutrients. The best studied such association is that between aphids and Buchnera aphidicola, which produces essential amino acids that are rare in the phloem sap diet. Genomic studies of Buchnera have provided a new means for inferring metabolic capabilities of the symbionts and their likely contributions to hosts. Despite evolutionary reduction of genome size, involving loss of most ancestral genes, Buchnera retains capabilities for biosynthesis of all essential amino acids. In contrast, most genes duplicating amino acid biosynthetic capabilities of hosts have been eliminated. In Buchnera of many aphids, genes for biosynthesis of leucine and tryptophan have been transferred from the chromosome to distinctive plasmids, a feature interpreted as a mechanism for overproducing these amino acids through gene amplification. However, the extent of plasmid-associated amplification varies between and within species, and plasmid-borne genes are sometimes fewer in number than single copy genes on the (polyploid) main chromosome. This supports the broader interpretation of the plasmid location as a means of achieving regulatory control of gene copy number and/or transcription. Buchnera genomes have eliminated most regulatory sequences, raising the question of the extent to which gene expression is moderated in response to changing demands imposed by host nutrition or other factors. Microarray analyses of the Buchnera transcriptome reveal only slight changes in expression of nutrition-related genes in response to shifts in host diet, with responses less dramatic than those observed for the related nonsymbiotic species, Escherichia coli. PMID- 14527995 TI - Distribution of NF-kappaB-binding sites across human chromosome 22. AB - We have mapped the chromosomal binding site distribution of a transcription factor in human cells. The NF-kappaB family of transcription factors plays an essential role in regulating the induction of genes involved in several physiological processes, including apoptosis, immunity, and inflammation. The binding sites of the NF-kappaB family member p65 were determined by using chromatin immunoprecipitation and a genomic microarray of human chromosome 22 DNA. Sites of binding were observed along the entire chromosome in both coding and noncoding regions, with an enrichment at the 5' end of genes. Strikingly, a significant proportion of binding was seen in intronic regions, demonstrating that transcription factor binding is not restricted to promoter regions. NF kappaB binding was also found at genes whose expression was regulated by tumor necrosis factor alpha, a known inducer of NF-kappaB-dependent gene expression, as well as adjacent to genes whose expression is not affected by tumor necrosis factor alpha. Many of these latter genes are either known to be activated by NF kappaB under other conditions or are consistent with NF-kappaB's role in the immune and apoptotic responses. Our results suggest that binding is not restricted to promoter regions and that NF-kappaB binding occurs at a significant number of genes whose expression is not altered, thereby suggesting that binding alone is not sufficient for gene activation. PMID- 14527996 TI - Mechanism of nucleotide incorporation opposite a thymine-thymine dimer by yeast DNA polymerase eta. AB - DNA polymerase eta (Poleta) has the unique ability to replicate through UV-light induced cyclobutane pyrimidine dimers. Here we use pre-steady-state kinetic analyses to examine the mechanism of nucleotide incorporation opposite a cis-syn thymine-thymine (TT) dimer and an identical nondamaged sequence by yeast Poleta. Poleta displayed "burst" kinetics for nucleotide incorporation opposite both the damaged and nondamaged templates. Although a slight decrease occurred in the affinity (Kd) of nucleotide binding opposite the TT dimer relative to the nondamaged template, the rate (kpol) of nucleotide incorporation was the same whether the template was damaged or nondamaged. These results strongly support a mechanism in which the nucleotide is directly inserted opposite the TT dimer by using its intrinsic base-pairing ability without any hindrance from the distorted geometry of the lesion. PMID- 14527997 TI - SecA2-dependent secretion of autolytic enzymes promotes Listeria monocytogenes pathogenesis. AB - Pathogenic bacteria secrete proteins that promote invasion of host tissues and resistance to immune responses. However, secretion mechanisms that contribute to the enormous morbidity and mortality of Gram-positive bacteria are largely undefined. An auxiliary protein secretion system (SecA2) has recently emerged in Listeria monocytogenes and eight other Gram-positive pathogens. Here, a proteomics approach identified seventeen SecA2-dependent secreted and surface proteins of L. monocytogenes, the two most abundant of which [the p60 and N acetylmuramidase (NamA) autolysins] hydrolyze bacterial peptidoglycan (PGN) and contribute to host colonization. SecA2-deficient (DeltaSecA2) bacteria were rapidly cleared after systemic infection of murine hosts, and in cultured cells showed reduced cell-cell spread. p60 or NamA deficiencies (Deltap60 and DeltaNamA) caused intermediate reductions in bacterial virulence in vivo, yet showed no defect for infection of cultured cells. Restoration of virulence in Deltap60 bacteria required full-length p60 with an intact catalytic domain, suggesting that PGN hydrolysis by p60 is crucial for L. monocytogenes virulence. Coordinated PGN hydrolysis by p60 and NamA activities is predicted to generate a muramyl glycopeptide, glucosaminylmuramyl dipeptide (GMDP), which is known to modify host inflammatory responses. Thus, SecA2-dependent secretion may promote release of muramyl peptides that subvert host pattern recognition. PMID- 14527998 TI - Gene expression profiling in Werner syndrome closely resembles that of normal aging. AB - Werner syndrome (WS) is a premature aging disorder, displaying defects in DNA replication, recombination, repair, and transcription. It has been hypothesized that several WS phenotypes are secondary consequences of aberrant gene expression and that a transcription defect may be crucial to the development of the syndrome. We used cDNA microarrays to characterize the expression of 6,912 genes and ESTs across a panel of 15 primary human fibroblast cell lines derived from young donors, old donors, and WS patients. Of the analyzed genes, 6.3% displayed significant differences in expression when either WS or old donor cells were compared with young donor cells. This result demonstrates that the WS transcription defect is specific to certain genes. Transcription alterations in WS were strikingly similar to those in normal aging: 91% of annotated genes displayed similar expression changes in WS and in normal aging, 3% were unique to WS, and 6% were unique to normal aging. We propose that a defect in the transcription of the genes as identified in this study could produce many of the complex clinical features of WS. The remarkable similarity between WS and normal aging suggests that WS causes the acceleration of a normal aging mechanism. This finding supports the use of WS as an aging model and implies that the transcription alterations common to WS and normal aging represent general events in the aging process. PMID- 14527999 TI - Microtubule destabilization and nuclear entry are sequential steps leading to toxicity in Huntington's disease. AB - There has been a longstanding debate regarding the role of proteolysis in Huntington's disease. The toxic peptide theory posits that N-terminal cleavage fragments of mutant Huntington's disease protein [mutant huntingtin (mhtt)] enter the nucleus to cause transcriptional dysfunction. However, recent data suggest a second model in which proteolysis of full-length mhtt is inhibited. Importantly, the two competing theories differ with respect to subcellular distribution of mhtt at initiation of toxicity: nuclear if cleaved and cytoplasmic in the absence of cleavage. Using quantitative single-cell analysis and time-lapse imaging, we show here that transcriptional dysfunction is "downstream" of cytoplasmic dysfunction. Primary and reversible toxic events involve destabilization of microtubules mediated by full-length mhtt before cleavage. Restoration of microtubule structure by taxol inhibits nuclear entry and increases cell survival. PMID- 14528001 TI - Rates of anterior tooth wear in Middle Pleistocene hominins from Sima de los Huesos (Sierra de Atapuerca, Spain). AB - This study presents quantitative data on the rates of anterior tooth wear in a Pleistocene human population. The data were obtained for the hominin sample of the Sima de los Huesos site in Atapuerca, Spain. The fossil record belongs to a minimum of 28 individuals of the same biological population, assigned to the species Homo heidelbergensis. We have estimated the original and the preserved crown height of the mandibular incisors (I1 and I2) of 11 individuals, whose age at death can be ascertained from the mineralization stage and tooth eruption. Results provide a range of 0.276-0.348 and 0.288-0.360 mm per year for the mean wear rate of the mandibular I1 and I2, respectively, in individuals approximately 16-18 years old. These data suggest that incisors' crowns would be totally worn out toward the fifth decade of life. Thus, we expect the life expectancy of this population to be seriously limited. These data, which could be contrasted with results obtained on hominins at other sites, could be of interest for estimating the death age of adult individuals. PMID- 14528000 TI - Functional map and domain structure of MET, the product of the c-met protooncogene and receptor for hepatocyte growth factor/scatter factor. AB - Little is known about the large ectodomain of MET, the product of the c-met protooncogene and receptor for hepatocyte growth factor/scatter factor (HGF/SF). Here, we establish by deletion mutagenesis that the HGF/SF and heparin-binding sites of MET are contained within a large N-terminal domain spanning the alpha chain (amino acids 25-307) and the first 212 amino acids of the beta-chain (amino acids 308-519). Neither the cystine-rich domain (amino acids 520-561) nor the C terminal half of MET (amino acids 562-932) bind HGF/SF or heparin directly. The MET ectodomain, which behaves as a rod-shaped monomer with a large Stokes radius in solution, binds HGF/SF in the absence or presence of heparin, and forms a stable HGF/SF-heparin-MET complex with 1:1:1 stoichiometry. We also show that the ligand-binding domain adopts a beta-propeller fold, which is similar to the N terminal domain of alphaV integrin, and that the C-terminal half contains four Ig domains (amino acids 563-654, 657-738, 742-836, and 839-924) of the unusual structural E set, which could be modeled on bacterial enzymes. Our studies provide 3D models and a functional map of the MET ectodomain. They have broad implications for structure-function of the MET receptor and the related semaphorin and plexin proteins. PMID- 14528002 TI - Rapid PCR amplification of minimal enediyne polyketide synthase cassettes leads to a predictive familial classification model. AB - A universal PCR method for the rapid amplification of minimal enediyne polyketide synthase (PKS) genes and the application of this methodology to clone remaining prototypical genes from producers of structurally determined enediynes in both family types are presented. A phylogenetic analysis of the new pool of bona fide enediyne PKS genes, consisting of three from 9-membered producers (neocarzinostatin, C1027, and maduropeptin) and three from 10-membered producers (calicheamicin, dynemicin, and esperamicin), reveals a clear genotypic distinction between the two structural families from which to form a predictive model. The results from this study support the postulation that the minimal enediyne PKS helps define the structural divergence of the enediyne core and provides the key tools for generating enediyne hybrid genes/molecular scaffolds; by using the model, a classification is also provided for the unknown enediyne PKS genes previously identified via genome scanning. PMID- 14528003 TI - Single-cell analysis of covalently closed circular DNA copy numbers in a hepadnavirus-infected liver. AB - Hepatitis B virus (hepadnavirus) infections are maintained by the presence of a small and regulated number of episomal viral genomes [covalently closed circular DNA (cccDNA)] in the nuclei of infected cells. Although a number of studies have measured the mean copy number of cccDNA molecules in hepadnaviral-infected cells, the distribution of individual copy numbers have not been reported. Using a PCR based assay, we examined the number of cccDNA molecules of the duck hepatitis B virus in single nuclei isolated from the liver of a chronically infected duck over the course of 131 days of infection. Nuclei were isolated from frozen serial biopsies and individually deposited into PCR microplates by flow sorting. Each nucleus was assayed by nested PCR for cccDNA and for cellular IFN-alpha genes as an internal control. We found that 90% of the nuclei assayed contained between 1 and 17 cccDNA molecules, with the remaining 10% containing more (90% confidence), and that differences in the mean number of copies and distribution of copy numbers occurred within the same animal at different times postinfection. Overall, the data suggest (i) that the number of cccDNA molecules per cell may fluctuate over time, and (ii) that, according to these fluctuations, a substantial fraction of cells may contain only one or a few copies. We infer from the results that infected hepatocytes express virus at different levels and that during cell division it is possible to segregate cells containing no cccDNA. PMID- 14528004 TI - Biomolecular hydration: from water dynamics to hydrodynamics. AB - Thermally driven rotational and translational diffusion of proteins and other biomolecules is governed by frictional coupling to their solvent environment. Prediction of this coupling from biomolecular structures is a longstanding biophysical problem, which cannot be solved without knowledge of water dynamics in an interfacial region comparable to the dry protein in volume. Efficient algorithms have been developed for solving the hydrodynamic equations of motion for atomic-resolution biomolecular models, but experimental diffusion coefficients can be reproduced only by postulating hundreds of rigidly bound water molecules. This static picture of biomolecular hydration is fundamentally inconsistent with magnetic relaxation dispersion experiments and molecular dynamics simulations, which both reveal a highly dynamic interface where rotation and exchange of nearly all water molecules are several orders of magnitude faster than biomolecular diffusion. Here, we resolve this paradox by means of a dynamic hydration model that explicitly links protein hydrodynamics to hydration dynamics. With the aid of this model, bona fide structure-based predictions of global biomolecular dynamics become possible, as demonstrated here for a set of 16 proteins for which accurate experimental rotational diffusion coefficients are available. PMID- 14528005 TI - Disruption of PPT2 in mice causes an unusual lysosomal storage disorder with neurovisceral features. AB - The palmitoyl protein thioesterase-2 (PPT2) gene encodes a lysosomal thioesterase homologous to PPT1, which is the enzyme defective in the human disorder called infantile neuronal ceroid lipofuscinosis. In this article, we report that PPT2 deficiency in mice causes an unusual form of neuronal ceroid lipofuscinosis with striking visceral manifestations. All PPT2-deficient mice displayed a neurodegenerative phenotype with spasticity and ataxia by 15 mo. The bone marrow was infiltrated by brightly autofluorescent macrophages and multinucleated giant cells, but interestingly, the macrophages did not have the typical appearance of foam cells commonly associated with other lysosomal storage diseases. Marked splenomegaly caused by extramedullary hematopoiesis was observed. The pancreas was grossly orange to brown as a result of massive storage of lipofuscin pigments in the exocrine (but not islet) cells. Electron microscopy showed that the storage material consisted of multilamellar membrane profiles ("zebra bodies"). In summary, PPT2 deficiency in mice manifests as a neurodegenerative disorder with visceral features. Although PPT2 deficiency has not been described in humans, manifestations would be predicted to include neurodegeneration with bone marrow histiocytosis, visceromegaly, brown pancreas, and linkage to chromosome 6p21.3 in affected families. PMID- 14528006 TI - Coupled prediction of protein secondary and tertiary structure. AB - The strong coupling between secondary and tertiary structure formation in protein folding is neglected in most structure prediction methods. In this work we investigate the extent to which nonlocal interactions in predicted tertiary structures can be used to improve secondary structure prediction. The architecture of a neural network for secondary structure prediction that utilizes multiple sequence alignments was extended to accept low-resolution nonlocal tertiary structure information as an additional input. By using this modified network, together with tertiary structure information from native structures, the Q3-prediction accuracy is increased by 7-10% on average and by up to 35% in individual cases for independent test data. By using tertiary structure information from models generated with the ROSETTA de novo tertiary structure prediction method, the Q3-prediction accuracy is improved by 4-5% on average for small and medium-sized single-domain proteins. Analysis of proteins with particularly large improvements in secondary structure prediction using tertiary structure information provides insight into the feedback from tertiary to secondary structure. PMID- 14528008 TI - Bias and error in estimates of equilibrium free-energy differences from nonequilibrium measurements. AB - In 1997, Jarzynski proved a remarkable equality that allows one to compute the equilibrium free-energy difference DeltaF between two states from the probability distribution of the nonequilibrium work W done on the system to switch between the states, e-DeltaF/kappaT =e-W/kappaT [Jarzynski, C. (1997) Phys. Rev. Lett. 87, 2690-2693]. The Jarzynski equality provides a powerful free-energy difference estimator from a set of N irreversible experiments and is closely related to free energy perturbation, a common computational technique for estimating free-energy differences. Despite the many applications of the Jarzynski estimator, its behavior is only poorly understood. In this article we derive the large N limit for the Jarzynski estimator bias, variance, and mean square error that is correct for arbitrary perturbations. We then analyze the properties of the Jarzynski estimator for all N when the probability distribution of work values is Gaussian, as occurs, for example, in the near-equilibrium regime. This allows us to quantitatively compare it to two other free-energy difference estimators: the mean work estimator and the fluctuation-dissipation theorem estimator. We show that, for near-equilibrium switching, the Jarzynski estimator is always superior to the mean work estimator and is even superior to the fluctuation-dissipation estimator for small N. The Jarzynski-estimator bias is shown to be the dominant source of error in many cases. Our expression for the bias is used to develop a bias-corrected Jarzynski free-energy difference estimator in the near-equilibrium regime. PMID- 14528009 TI - Information exchange and the robustness of organizational networks. AB - The dynamics of information exchange is an important but understudied aspect of collective communication, coordination, and problem solving in a wide range of distributed systems, both physical (e.g., the Internet) and social (e.g., business firms). In this paper, we introduce a model of organizational networks according to which links are added incrementally to a hierarchical backbone and test the resulting networks under variable conditions of information exchange. Our main result is the identification of a class of multiscale networks that reduce, over a wide range of environments, the likelihood that individual nodes will suffer congestion-related failure and that the network as a whole will disintegrate when failures do occur. We call this dual robustness property of multiscale networks "ultrarobustness." Furthermore, we find that multiscale networks attain most of their robustness with surprisingly few link additions, suggesting that ultrarobust organizational networks can be generated in an efficient and scalable manner. Our results are directly relevant to the relief of congestion in communication networks and also more broadly to activities, like distributed problem solving, that require individuals to exchange information in an unpredictable manner. PMID- 14528007 TI - Protective immunity against Helicobacter is characterized by a unique transcriptional signature. AB - Immunization with a whole-cell sonicate vaccine of Helicobacter felis in conjunction with cholera toxin as a mucosal adjuvant induces long-term protective immunity in a majority of laboratory mice. We have combined gene expression profiling and immunohistochemical analysis on a set of immunized animals to better understand the mechanism of protection. The stomachs of protected animals exhibited a strikingly different transcriptional profile compared with those of nonprotected or control mice, indicating that vaccination targets the appropriate site and leaves a molecular signature. Among the genes whose up-regulation is significantly correlated with protection are a number of adipocyte-specific factors. These include the fat-cell-specific cytokines adipsin, resistin, and adiponectin and the adipocyte surface marker CD36. Interestingly, potentially protective T and B lymphocytes can be found embedded in the adipose tissue surrounding protected stomachs but never in control or unprotected stomachs. Adipsin-specific immunohistochemical staining of protected stomach sections further revealed molecular cross-talk between adjacent lymphoid and adipose cell populations. We propose a mechanism of protection that involves the effector responses of either or both lymphocyte subclasses as well as the previously unappreciated paracrine functions of adipose tissue surrounding the resident lymphocytes. PMID- 14528010 TI - Slx1-Slx4 are subunits of a structure-specific endonuclease that maintains ribosomal DNA in fission yeast. AB - In most eukaryotes, genes encoding ribosomal RNAs (rDNA) are clustered in long tandem head-to-tail repeats. Studies of Saccharomyces cerevisiae have indicated that rDNA copy number is maintained through recombination events associated with site-specific blockage of replication forks (RFs). Here, we describe two Schizosaccharomyces pombe proteins, homologs of S. cerevisiae Slx1 and Slx4, as subunits of a novel type of endonuclease that maintains rDNA copy number. The Slx1-Slx4-dependent endonuclease introduces single-strand cuts in duplex DNA on the 3' side of junctions with single-strand DNA. Deletion of Slx1 or Rqh1 RecQ like DNA helicase provokes rDNA contraction, whereas simultaneous elimination of Slx1-Slx4 endonuclease and Rqh1 is lethal. Slx1 associates with chromatin at two foci characteristic of the two rDNA repeat loci in S. pombe. We propose a model in which the Slx1-Slx4 complex is involved in the control of the expansion and contraction of the rDNA loci by initiating recombination events at stalled RFs. PMID- 14528011 TI - Telomerase RNA accumulates in Cajal bodies in human cancer cells. AB - Telomerase synthesizes telomeric DNA repeats at the ends of eukaryotic chromosomes. The RNA component of the enzyme (hTR) provides the template for telomere synthesis, which is catalyzed by telomerase reverse transcriptase (hTERT). Little is known regarding the subcellular localization of hTR and hTERT and the pathway by which telomerase is assembled. Here we report the first glimpse of the detailed subcellular localization of endogenous hTR in human cells, which we obtained by fluorescence in situ hybridization (FISH). Our studies have revealed a distinctive hTR localization pattern in cancer cells. We have found that hTR accumulates within intranuclear foci called Cajal bodies in all typical tumor-derived cell lines examined (in which telomerase is active), but not in primary or ALT cells (where little or no hTERT is present). Accumulation of hTR in the Cajal bodies of primary cells is induced when hTERT is ectopically expressed. Moreover, we report that hTERT is also found in Cajal bodies. Our data suggest that Cajal bodies are involved in the assembly and/or function of human telomerase. PMID- 14528012 TI - The chromokinesin, KLP3A, dives mitotic spindle pole separation during prometaphase and anaphase and facilitates chromatid motility. AB - Mitosis requires the concerted activities of multiple microtubule (MT)-based motor proteins. Here we examined the contribution of the chromokinesin, KLP3A, to mitotic spindle morphogenesis and chromosome movements in Drosophila embryos and cultured S2 cells. By immunofluorescence, KLP3A associates with nonfibrous punctae that concentrate in nuclei and display MT-dependent associations with spindles. These punctae concentrate in indistinct domains associated with chromosomes and central spindles and form distinct bands associated with telophase midbodies. The functional disruption of KLP3A by antibodies or dominant negative proteins in embryos, or by RNA interference (RNAi) in S2 cells, does not block mitosis but produces defects in mitotic spindles. Time-lapse confocal observations of mitosis in living embryos reveal that KLP3A inhibition disrupts the organization of interpolar (ip) MTs and produces short spindles. Kinetic analysis suggests that KLP3A contributes to spindle pole separation during the prometaphase-to-metaphase transition (when it antagonizes Ncd) and anaphase B, to normal rates of chromatid motility during anaphase A, and to the proper spacing of daughter nuclei during telophase. We propose that KLP3A acts on MTs associated with chromosome arms and the central spindle to organize ipMT bundles, to drive spindle pole separation and to facilitate chromatid motility. PMID- 14528013 TI - Scavenger receptor BI (SR-BI) clustered on microvillar extensions suggests that this plasma membrane domain is a way station for cholesterol trafficking between cells and high-density lipoprotein. AB - Receptor-mediated trafficking of cholesterol between lipoproteins and cells is a fundamental biological process at the organismal and cellular levels. In contrast to the well-studied pathway of LDL receptor-mediated endocytosis, little is known about the trafficking of high-density lipoprotein (HDL) cholesterol by the HDL receptor, scavenger receptor BI (SR-BI). SR-BI mediates HDL cholesteryl ester uptake in a process in which HDL lipids are selectively transferred to the cell membrane without the uptake and degradation of the HDL particle. We report here the cell surface locale where the trafficking of HDL cholesterol occurs. Fluorescence confocal microscopy showed SR-BI in patches and small extensions of the cell surface that were distinct from sites of caveolin-1 expression. Electron microscopy showed SR-BI in patches or clusters primarily on microvillar extensions of the plasma membrane. The organization of SR-BI in this manner suggests that this microvillar domain is a way station for cholesterol trafficking between HDL and cells. The types of phospholipids in this domain are unknown, but SR-BI is not strongly associated with classical membrane rafts rich in detergent-resistant saturated phospholipids. We speculate that SR-BI is in a more fluid membrane domain that will favor rapid cholesterol flux between the membrane and HDL. PMID- 14528014 TI - The adaptor protein ARH escorts megalin to and through endosomes. AB - Megalin is an endocytic receptor that binds multiple ligands and is essential for many physiological processes such as brain development and uptake of proteins by the kidney tubule, yolk sac, and thyroid. The cytoplasmic tail of megalin contains two FXNPXY motifs. Autosomal recessive hypercholesterolemia (ARH) is an adaptor protein that binds to the FXNPXY motif of the low-density lipoprotein receptor as well as clathrin and AP-2. We found that ARH also binds to the first FXNPXY motif of megalin in two-hybrid, pull-down and coimmunoprecipitation assays. ARH colocalizes with megalin in clathrin coated pits and in recycling endosomes in the Golgi region. When cells are treated with nocodazole, the recycling endosomes containing megalin and ARH disperse. On internalization of megalin, ARH and megalin are first seen in clathrin coated pits followed by sequential localization in early endosomes and tubular recycling endosomes in the pericentriolar region followed by their reappearance at the cell surface. Expression of ARH in Madin-Darby canine kidney cells expressing megalin mini receptors enhances megalin-mediated uptake of 125I-lactoferrin, a megalin ligand. These results show that ARH facilitates endocytosis of megalin, escorts megalin along its endocytic route and raise the possibility that transport through the endosomal system is selective and requires interaction with specific adaptor proteins. PMID- 14528015 TI - Synaptophysin I controls the targeting of VAMP2/synaptobrevin II to synaptic vesicles. AB - Synaptic vesicle (SV) proteins are synthesized at the level of the cell body and transported down the axon in membrane precursors of SVs. To investigate the mechanisms underlying sorting of proteins to SVs, fluorescent chimeras of vesicle associated membrane protein (VAMP) 2, its highly homologous isoform VAMP1 and synaptotagmin I (SytI) were expressed in hippocampal neurons in culture. Interestingly, the proteins displayed a diffuse component of distribution along the axon. In addition, VAMP2 was found to travel in vesicles that constitutively fuse with the plasma membrane. Coexpression of VAMP2 with synaptophysin I (SypI), a major resident of SVs, restored the correct sorting of VAMP2 to SVs. The effect of SypI on VAMP2 sorting was dose dependent, being reversed by increasing VAMP2 expression levels, and highly specific, because the sorting of the SV proteins VAMP1 and SytI was not affected by SypI. The cytoplasmic domain of VAMP2 was found to be necessary for both the formation of VAMP2-SypI hetero-dimers and for VAMP2 sorting to SVs. These data support a role for SypI in directing the correct sorting of VAMP2 in neurons and demonstrate that a direct interaction between the two proteins is required for SypI in order to exert its effect. PMID- 14528016 TI - Dynamic and regulated association of caveolin with lipid bodies: modulation of lipid body motility and function by a dominant negative mutant. AB - Caveolins are a crucial component of caveolae but have also been localized to the Golgi complex, and, under some experimental conditions, to lipid bodies (LBs). The physiological relevance and dynamics of LB association remain unclear. We now show that endogenous caveolin-1 and caveolin-2 redistribute to LBs in lipid loaded A431 and FRT cells. Association with LBs is regulated and reversible; removal of fatty acids causes caveolin to rapidly leave the lipid body. We also show by subcellular fractionation, light and electron microscopy that during the first hours of liver regeneration, caveolins show a dramatic redistribution from the cell surface to the newly formed LBs. At later stages of the regeneration process (when LBs are still abundant), the levels of caveolins in LBs decrease dramatically. As a model system to study association of caveolins with LBs we have used brefeldin A (BFA). BFA causes rapid redistribution of endogenous caveolins to LBs and this association was reversed upon BFA washout. Finally, we have used a dominant negative LB-associated caveolin mutant (cavDGV) to study LB formation and to examine its effect on LB function. We now show that the cavDGV mutant inhibits microtubule-dependent LB motility and blocks the reversal of lipid accumulation in LBs. PMID- 14528018 TI - Vacuole size control: regulation of PtdIns(3,5)P2 levels by the vacuole associated Vac14-Fig4 complex, a PtdIns(3,5)P2-specific phosphatase. AB - In the budding yeast Saccharomyces cerevisiae, phosphatidylinositol 3,5 bisphosphate (PtdIns(3,5)P2) is synthesized by a single phosphatidylinositol 3 phosphate 5-kinase, Fab1. Cells deficient in PtdIns(3,5)P2 synthesis exhibit a grossly enlarged vacuole morphology, whereas increased levels of PtdIns(3,5)P2 provokes the formation of multiple small vacuoles, suggesting a specific role for PtdIns(3,5)P2 in vacuole size control. Genetic studies have indicated that Fab1 kinase is positively regulated by Vac7 and Vac14; deletion of either gene results in ablation of PtdIns(3,5)P2 synthesis and the formation of a grossly enlarged vacuole. More recently, a suppressor of vac7Delta mutants was identified and shown to encode a putative phosphoinositide phosphatase, Fig4. We demonstrate that Fig4 is a magnesium-activated PtdIns(3,5)P2-selective phosphoinositide phosphatase in vitro. Analysis of a Fig4-GFP fusion protein revealed that the Fig4 phosphatase is localized to the limiting membrane of the vacuole. Surprisingly, in the absence of Vac14, Fig4-GFP no longer localizes to the vacuole. However, Fig4-GFP remains localized to the grossly enlarged vacuoles of vac7 deletion mutants. Consistent with these observations, we found that Fig4 physically associates with Vac14 in a common membrane-associated complex. Our studies indicate that Vac14 both positively regulates Fab1 kinase activity and directs the localization/activation of the Fig4 PtdIns(3,5)P2 phosphatase. PMID- 14528017 TI - Endocytosis of epithelial apical junctional proteins by a clathrin-mediated pathway into a unique storage compartment. AB - The adherens junction (AJ) and tight junction (TJ) are key regulators of epithelial polarity and barrier function. Loss of epithelial phenotype is accompanied by endocytosis of AJs and TJs via unknown mechanisms. Using a model of calcium depletion, we defined the pathway of internalization of AJ and TJ proteins (E-cadherin, p120 and beta-catenins, occludin, JAM-1, claudins 1 and 4, and ZO-1) in T84 epithelial cells. Proteinase protection assay and immunocytochemistry revealed orchestrated internalization of AJs and TJs into a subapical cytoplasmic compartment. Disruption of caveolae/lipid rafts did not prevent endocytosis, nor did caveolin-1 colocalize with internalized junctional proteins. Furthermore, AJ and TJ proteins did not colocalize with the macropinocytosis marker dextran. Inhibitors of clathrin-mediated endocytosis blocked internalization of AJs and TJs, and junctional proteins colocalized with clathrin and alpha-adaptin. AJ and TJ proteins were observed to enter early endosomes followed by movement to organelles that stained with syntaxin-4 but not with markers of late and recycling endosomes, lysosomes, or Golgi. These results indicate that endocytosis of junctional proteins is a clathrin-mediated process leading into a unique storage compartment. Such mechanisms may mediate the disruption of intercellular contacts during normal tissue remodeling and in pathology. PMID- 14528019 TI - Roles of phosphoinositides and of Spo14p (phospholipase D)-generated phosphatidic acid during yeast sporulation. AB - During yeast sporulation, internal membrane synthesis ensures that each haploid nucleus is packaged into a spore. Prospore membrane formation requires Spo14p, a phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2]-stimulated phospholipase D (PLD), which hydrolyzes phosphatidylcholine (PtdCho) to phosphatidic acid (PtdOH) and choline. We found that both meiosis and spore formation also require the phosphatidylinositol (PtdIns)/PtdCho transport protein Sec14p. Specific ablation of the PtdIns transport activity of Sec14p was sufficient to impair spore formation but not meiosis. Overexpression of Pik1p, a PtdIns 4-kinase, suppressed the sec14-1 meiosis and spore formation defects; conversely, pik1-ts diploids failed to undergo meiosis and spore formation. The PtdIns(4)P 5-kinase, Mss4p, also is essential for spore formation. Use of phosphoinositide-specific GFP-PH domain reporters confirmed that PtdIns(4,5)P2 is enriched in prospore membranes. sec14, pik1, and mss4 mutants displayed decreased Spo14p PLD activity, whereas absence of Spo14p did not affect phosphoinositide levels in vivo, suggesting that formation of PtdIns(4,5)P2 is important for Spo14p activity. Spo14p-generated PtdOH appears to have an essential role in sporulation, because treatment of cells with 1-butanol, which supports Spo14p-catalyzed PtdCho breakdown but leads to production of Cho and Ptd-butanol, blocks spore formation at concentrations where the inert isomer, 2-butanol, has little effect. Thus, rather than a role for PtdOH in stimulating PtdIns(4,5)P2 formation, our findings indicate that during sporulation, Spo14p-mediated PtdOH production functions downstream of Sec14p-, Pik1p-, and Mss4p-dependent PtdIns(4,5)P2 synthesis. PMID- 14528020 TI - Direct binding of the ligand PSG17 to CD9 requires a CD9 site essential for sperm egg fusion. AB - The function currently attributed to tetraspanins is to organize molecular complexes in the plasma membrane by using multiple cis-interactions. Additionally, the tetraspanin CD9 may be a receptor that binds the soluble ligand PSG17, a member of the immunoglobulin superfamily (IgSF)/CEA subfamily. However, previous data are also consistent with the PSG17 receptor being a CD9 cis associated protein. In the current study, CD9 extracellular loop (EC2) specifically bound to PSG17-coated beads, indicating a direct interaction between the two proteins. However, CD9-EC2 did not bind to PSG17-coated beads if the CD9 EC2 had the mutation SFQ (173-175) to AAA, a previously studied mutation in egg CD9 that abolishes sperm-egg fusion. Also, PSG17 bound to 293 T cells transfected with wild-type CD9 but not the mutant CD9. By immunofluorescence, PSG17 bound to wild-type eggs but not to CD9 null eggs. The presence of approximately 2 microM recombinant PSG17 produced a significant and reversible inhibition (60-80%) of sperm-egg fusion. Thus, we conclude that CD9 is a receptor for PSG17 and when the PSG17 binding site is mutated or occupied, sperm-egg fusion is impaired. These findings suggest that egg CD9 may function in gamete fusion by binding to a sperm IgSF/CEA subfamily member and such proteins have previously been identified on sperm. PMID- 14528021 TI - Tyrosine phosphatase epsilon is a positive regulator of osteoclast function in vitro and in vivo. AB - Protein tyrosine phosphorylation is a major regulator of bone metabolism. Tyrosine phosphatases participate in regulating phosphorylation, but roles of specific phosphatases in bone metabolism are largely unknown. We demonstrate that young (<12 weeks) female mice lacking tyrosine phosphatase epsilon (PTPepsilon) exhibit increased trabecular bone mass due to cell-specific defects in osteoclast function. These defects are manifested in vivo as reduced association of osteoclasts with bone and as reduced serum concentration of C-terminal collagen telopeptides, specific products of osteoclast-mediated bone degradation. Osteoclast-like cells are generated readily from PTPepsilon-deficient bone-marrow precursors. However, cultures of these cells contain few mature, polarized cells and perform poorly in bone resorption assays in vitro. Podosomes, structures by which osteoclasts adhere to matrix, are disorganized and tend to form large clusters in these cells, suggesting that lack of PTPepsilon adversely affects podosomal arrangement in the final stages of osteoclast polarization. The gender and age specificities of the bone phenotype suggest that it is modulated by hormonal status, despite normal serum levels of estrogen and progesterone in affected mice. Stimulation of bone resorption by RANKL and, surprisingly, Src activity and Pyk2 phosphorylation are normal in PTPepsilon-deficient osteoclasts, indicating that loss of PTPepsilon does not cause widespread disruption of these signaling pathways. These results establish PTPepsilon as a phosphatase required for optimal structure, subcellular organization, and function of osteoclasts in vivo and in vitro. PMID- 14528022 TI - Targeting of a tropomyosin isoform to short microfilaments associated with the Golgi complex. AB - A growing body of evidence suggests that the Golgi complex contains an actin based filament system. We have previously reported that one or more isoforms from the tropomyosin gene Tm5NM (also known as gamma-Tm), but not from either the alpha- or beta-Tm genes, are associated with Golgi-derived vesicles (Heimann et al., (1999). J. Biol. Chem. 274, 10743-10750). We now show that Tm5NM-2 is sorted specifically to the Golgi complex, whereas Tm5NM-1, which differs by a single alternatively spliced internal exon, is incorporated into stress fibers. Tm5NM-2 is localized to the Golgi complex consistently throughout the G1 phase of the cell cycle and it associates with Golgi membranes in a brefeldin A-sensitive and cytochalasin D-resistant manner. An actin antibody, which preferentially reacts with the ends of microfilaments, newly reveals a population of short actin filaments associated with the Golgi complex and particularly with Golgi-derived vesicles. Tm5NM-2 is also found on these short microfilaments. We conclude that an alternative splice choice can restrict the sorting of a tropomyosin isoform to short actin filaments associated with Golgi-derived vesicles. Our evidence points to a role for these Golgi-associated microfilaments in vesicle budding at the level of the Golgi complex. PMID- 14528023 TI - A novel cytoplasmic tail MXXXL motif mediates the internalization of prostate specific membrane antigen. AB - Prostate-specific membrane antigen (PSMA) is a transmembrane protein expressed at high levels in prostate cancer and in tumor-associated neovasculature. In this study, we report that PSMA is internalized via a clathrin-dependent endocytic mechanism and that internalization of PSMA is mediated by the five N-terminal amino acids (MWNLL) present in its cytoplasmic tail. Deletion of the cytoplasmic tail abolished PSMA internalization. Mutagenesis of N-terminal amino acid residues at position 2, 3, or 4 to alanine did not affect internalization of PSMA, whereas mutation of amino acid residues 1 or 5 to alanine strongly inhibited internalization. Using a chimeric protein composed of Tac antigen, the alpha-chain of interleukin 2-receptor, fused to the first five amino acids of PSMA (Tac-MWNLL), we found that this sequence is sufficient for PSMA internalization. In addition, inclusion of additional alanines into the MWNLL sequence either in the Tac chimera or the full-length PSMA strongly inhibited internalization. From these results, we suggest that a novel MXXXL motif in the cytoplasmic tail mediates PSMA internalization. We also show that dominant negative micro2 of the adaptor protein (AP)-2 complex strongly inhibits the internalization of PSMA, indicating that AP-2 is involved in the internalization of PSMA mediated by the MXXXL motif. PMID- 14528024 TI - Klotho, a gene related to a syndrome resembling human premature aging, functions in a negative regulatory circuit of vitamin D endocrine system. AB - The klotho gene encodes a novel type I membrane protein of beta-glycosidase family and is expressed principally in distal tubule cells of the kidney and choroid plexus in the brain. These mutants displayed abnormal calcium and phosphorus homeostasis together with increased serum 1,25-(OH)2D. In kl-/- mice at the age of 3 wk, elevated levels of serum calcium (10.9 +/- 0.31 mg/dl vs. 10.0 +/- 0.048 mg/dl in wild-type mice), phosphorus (14.7 +/- 1.1 mg/dl vs. 9.7 +/- 1.5 mg/dl in wild type) and most notably, 1,25-(OH)2D (403 +/- 99.7 mg/dl vs. 88.0 +/- 34.0 mg/dl in wild type) were observed. Reduction of serum 1,25-(OH)2D concentrations by dietary restriction resulted in alleviation of most of the phenotypes, suggesting that they are downstream events resulting from elevated 1,25-(OH)2D. We searched for the signals that lead to up-regulation of vitamin D activating enzymes. We examined the response of 1alpha-hydroxylase gene expression to calcium regulating hormones, such as PTH, calcitonin, and 1,25 (OH)2D3. These pathways were intact in klotho null mutant mice, suggesting the existence of alternate regulatory circuits. We also found that the administration of 1,25-(OH)2D3 induced the expression of klotho in the kidney. These observations suggest that klotho may participate in a negative regulatory circuit of the vitamin D endocrine system, through the regulation of 1alpha-hydroxylase gene expression. PMID- 14528026 TI - Contribution of ionic currents to magnetoencephalography (MEG) and electroencephalography (EEG) signals generated by guinea-pig CA3 slices. AB - A mathematical model was used to analyse the contributions of different types of ionic currents in the pyramidal cells of longitudinal CA3 slices to the magnetic fields and field potentials generated by this preparation. Murakami et al. recently showed that a model based on the work of Traub et al. provides a quantitatively accurate account of the basic features of three types of empirical data (magnetic fields outside the slice, extracellular field potentials within the slice and intracellular potentials within the pyramidal neurons) elicited by stimulations of the soma and apical dendrites. This model was used in the present study to compute the net current dipole moment (Q) due to each of the different voltage- and ligand-gated channels in the cells in the presence of fast GABAA inhibition. These values of Q are proportional to the magnetic field and electrical potential far away from the slice. The intrinsic conductances were found to be more important than the synaptic conductances in determining the shape and magnitude of Q. Among the intrinsic conductances, the sodium (gNa) and delayed-rectifier potassium (gK(DR)) channels were found to produce sharp spikes. The high-threshold calcium channel (gCa) and C-type potassium channel (gK(C)) primarily determined the overall current waveforms. The roles of gCa and gK(C) were independent of small perturbations in these channel densities in the apical and basal dendrites. A combination of gNa, gK(DR), gCa, and gK(C) accounted for most of the evoked responses, except for later slow components, which were primarily due to synaptic channels. PMID- 14528025 TI - Developmental aspects of spinal locomotor function: insights from using the in vitro mouse spinal cord preparation. AB - Over the last five years, rapid advances have been made in our understanding of the location, function, and recently, organization of the central pattern generator (CPG) for locomotion. In the mammal, the use of the neonatal rat has largely contributed to these advances. Additionally, the use of the in vitro mouse spinal cord preparation is becoming more common, catalysed in part by the potential for the use of genetic approaches to study locomotor function. Although tempting, it is necessary to resist the a priori assumption that the organization of the spinal CPG is identical in the rat and mouse. This review will describe the development of locomotor-like behaviour in the mouse from embryonic day 12 to postnatal day 14. While there are still many gaps in our knowledge, compared with the rat, the in vitro mouse appears to follow a qualitatively similar course of locomotor development. The emphasis in this review is the use or potential use of the mouse as a complement to existing data using the neonatal rat preparation. PMID- 14528028 TI - Exercise-induced changes in plasma composition increase erythrocyte Na+,K+ ATPase, but not Na+-K+-2Cl- cotransporter, activity to stimulate net and unidirectional K+ transport in humans. AB - We tested the hypothesis that exercise-induced changes in plasma composition result in peak stimulation of erythrocyte unidirectional K+ (JK,in) and net K+ (JK,net) transport within the first 120 s. In experimental series 1 (7 men; 2 women), plasma [K+] was continuously measured in vitro (37 degrees C) after the addition of red blood cells (RBCs) obtained from rested subjects (resting RBCs) into an exercise-simulated plasma (ESP; increased plasma osmolality, [K+], [H+], [lactate] and [adrenaline] (epinephrine)), and JK,net calculated. In experimental series 2 (7 men; 4 women), resting RBCs were incubated in true exercise plasma (TEP) obtained after two 30 s bouts of high intensity leg cycling exercise to determine JK,net and JK,in (via RBC 86Rb accumulation). JK,net of resting RBCs increased from 0.9 +/- 28.7 in resting plasma to 285 +/- 164 mmol (l RBCs)-1 h-1 in ESP and to 178 +/- 60 mmol (l RBCs)-1 h-1 after 10 s in TEP. Both JK,net and JK,in peaked within 10 s of incubation and decreased rapidly during the initial 120 s. The use of inhibitors for the Na+,K+-ATPase (ouabain) and the Na+-K+-2Cl- cotransporter (NKCC; bumetanide) indicated that rapid increases in JK,in and JK,net upon incubation of resting RBCs in TEP were due primarily to increased Na+,K+-ATPase activity; the NKCC appeared to be involved only when the Na+,K+ ATPase was blocked. It is concluded that RBCs rapidly increase JK,in and JK,net in response to exercise-induced changes in plasma composition. PMID- 14528029 TI - On the selective advantage of suicide. PMID- 14528027 TI - Denervation of carotid baro- and chemoreceptors in humans. AB - Experimental denervation in animals has shown that carotid baro- and chemoreceptors play an eminent role in maintaining blood pressure and blood gas homeostasis. Denervation of carotid sinus baro- and chemoreceptors in humans may occur as a complication of invasive interventions on the neck or after experimental surgical treatment in asthma. In this topical review, the short- and long-term effects of carotid baro- and chemoreceptor denervation on the control of circulation and ventilation in humans are discussed. Carotid baroreceptor denervation in humans causes a persistent decrease in vagal and sympathetic baroreflex sensitivity and an increase in blood pressure variability; however, carotid denervation does not lead to chronic hypertension. Therefore, although carotid baroreceptors contribute to short-term blood pressure control, other receptors are able to maintain normal chronic blood pressure levels in the absence of carotid baroreceptors. Conversely, carotid chemoreceptor denervation leads to permanent abolition of normocapnic ventilatory responses to hypoxia and reduced ventilatory responses to hypercapnia. PMID- 14528030 TI - Cell senescence: an evaluation of replicative senescence in culture as a model for cell aging in situ. PMID- 14528032 TI - Effect of DNA repair on aging of transgenic Drosophila melanogaster: I. mei-41 locus. AB - Aging appears to be increased by diminished DNA repair. To study this relationship between aging and DNA repair, we measured the life span of Drosophila melanogaster males in the absence of mei-41 excision repair and transgenic flies with 1 or 2 extra copies of the mei-41 wild-type gene. Life span was significantly reduced in the absence of repair and was significantly increased by an extra dose of excision repair. However, these changes in life span with alterations in DNA repair were not large. PMID- 14528033 TI - Effect of DL-alpha-lipoic acid on the status of lipid peroxidation and lipids in aged rats. AB - The effect of dextro and levo (DL)-alpha-lipoic acid on lipid peroxidation and lipids has been evaluated in plasma, liver, and kidney of young and aged rats. The levels of thiobarbituric acid reactive substances (TBARS) and lipids were considerably higher in aged rats compared with younger controls. DL-alpha-lipoic acid (100 mg/kg body wt/day) was administered intraperitoneally for 7 and 14 days. Supplementation of lipoic acid in aged rats prevents the elevated levels of TBARS and lipids. From our observations, we conclude that lipoic acid is very effective in normalizing age-related alterations in lipids, and it can be implemented in the aged to minimize age-associated disorders where free radicals are the major cause. PMID- 14528034 TI - Effect of aging on rat tissue peptidase activities. AB - The process of aging is known to involve alterations in the activity of peptidases and proteases. However, the precise changes in the activity of many peptidases in aged tissues have not yet been fully characterized, and both decreases and increases in both peptidase activity and peptide levels have been reported to occur during the aging process. In the present study, we measured the activity of several peptidases in selected tissues (brain cortex, brain stem, liver, kidney, heart, and lung) of the young adult (3 months old) and aged (18 months old and 22 months old) rat. The activities of prolyl endopeptidase, pyroglutamyl peptidase I, puromycin sensitive aminopeptidase, and aminopeptidase N were assayed using beta-naphthylamine aminoacidic derivatives as substrates. The activity of the soluble fractions of prolyl endopeptidase was found to be reduced in the lungs of aged animals, while reduced activity of soluble pyroglutamyl peptidase I and also aminopeptidase N was measured in the aged kidney and heart, respectively. In contrast, increased activity of particulate prolyl endopeptidase was measured in the brain stem of older animals. Since most of these changes can be correlated with known alterations in the levels of peptides controlled by each enzyme, the results of the present study indicate that the studied peptidases may play an important role in regulating tissue peptide levels during aging. PMID- 14528035 TI - Inhibited angiogenesis in aging: a role for TIMP-2. AB - Factors responsible for age-associated impairment of angiogenesis are poorly understood. We observed that in aged mice, new fibrovascular tissue within subcutaneous polyvinyl alcohol sponges expressed more tissue inhibitor of metalloproteinases (TIMP)-2 than did corresponding tissue from young mice. In complementary studies in vitro, we utilized young and aged human microvascular endothelial cell lines (hmEC36 and hmEC90, respectively) and compared their morphogenetic capacity within three-dimensional collagen. HmEC90 exhibited poor formation of tubular, capillary-like structures in vitro, diminished expression of active matrix metalloproteinase (MMP)-2, and similar or lesser amounts of MT1 MMP relative to hmEC36. Correspondingly, the MMP inhibitor GM6001 decreased tubulogenesis by hmEC36 to levels observed for hmEC90. In vitro, hmEC90 expressed similar quantities of TIMP-1, but more TIMP-2 than did hmEC36. Accordingly, purified TIMP-2 inhibited tubulogenesis by hmEC36. Collectively, our studies indicate that elevated levels of TIMP-2 modulate decreased angiogenesis in aged tissues, most likely via TIMP-2-mediated inhibition of MMP-2 and MT1-MMP. PMID- 14528036 TI - Geriatricians, continuous quality improvement, and improved care for older persons. PMID- 14528037 TI - Patient safety in geriatrics: a call for action. AB - Patient safety has become a major public health concern following the publication of the landmark report, To Err Is Human, by the Institute of Medicine in 1999. This report, along with a subsequent report, Crossing the Quality Chasm, recommended the design of a safer health care system by integrating well established safety methods to avert medical errors. However, neither patient safety report specifically addressed the implications of safety for elderly patients. This article examines those implications by describing the association between aging and medical errors, identifying geriatric syndromes as medical errors, and focusing on six recommendations that will improve the safety of geriatric care. These six recommendations include the detection and reporting of geriatric syndromes, identifying system failures when geriatric syndromes occur, establishing dedicated geriatric units, improving the continuity of care, reducing adverse drug events, and improving geriatric training programs. PMID- 14528038 TI - Future history: medical informatics in geriatrics. AB - With deference to Isaac Asimov's The Foundation, which is the inspiration for this series, we briefly describe the "present history" of medical informatics (the application of information technology in medicine) in geriatrics, and then project a "future history" of this same endeavor. The older patient often has multiple acute and chronic problems that require management by a variety of medical professionals in a variety of settings. Proper care necessitates efficient gathering, integration, and management of information by each professional in each setting. As medical informatics evolves, we project that barriers to information exchange (both between providers and between providers and patients) will continue to decrease while the quality and relevance of exchanged information will continue to increase. The nexus of care will be the electronic medical record (EMR), which will shed its current paper chart metaphor and adopt an industrial process metaphor based on tasks and tolerances or goals. The multidisciplinary management of geriatric patients will strike a new balance: doctors, nurses, allied health professionals, family, and patients will all participate in the management of the patient's care. The EMR will coordinate data from a variety of novel sources, including wearable sensors monitoring physiologic parameters, falls, diet, ambulation, and medication compliance. The highly organized data in the EMR will allow explicit decision support for computer-facilitated, evidence-based care; will empower midlevel providers and patients with an increased role in the care plan; and will promote the realignment of care from hospitals/clinics to the patient's home. PMID- 14528039 TI - A consensus statement on nonemergent medical transportation services for older persons. PMID- 14528040 TI - Visual attention and driving behaviors among community-living older persons. AB - BACKGROUND: Older drivers have higher rates of crashes per mile driven compared with most other drivers, and these crashes result in greater morbidity and mortality. Various aspects of cognition, particularly visual attention, have been linked with crash risk among older individuals. The current study was designed to specify those cognitive variables associated with specific on-road driving behaviors in a sample of older, nonclinic-referred individuals. METHODS: 35 community-residing active drivers aged 72 years and older (M = 80) underwent a standardized, on-road driving evaluation involving parking lot maneuvers, and urban, suburban, and highway driving. They were also administered tests of visual attention, executive function, visuospatial cognition, and memory. RESULTS: Driving score was significantly correlated with visual attention, visual memory, and executive function. Visual attention was associated with 25 of 36 driving behaviors, including those involving scanning the environment, interaction with traffic or pedestrians, and distance judgments. Executive function and visual memory were associated with fewer maneuvers, most of which were a subset of maneuvers that correlated with visual attention. CONCLUSIONS: Visual attention, a cognitive function involving search, selection, and switching, plays an important role in driving risk among older drivers. In the current study, key driving maneuvers involving interaction with other vehicles/pedestrians, such as yielding right of way and negotiating safe turns or merges, have the greatest association with visual attention. Specification of both the cognitive risk factors and their impact on problematic driving maneuvers may provide guidelines for developing targeted interventions to reduce risk among older adults. PMID- 14528041 TI - Management of behavioral symptoms in progressive degenerative dementias. AB - Management of behavioral symptoms of Alzheimer's disease and other progressive degenerative dementias poses continuous challenge to both family and professional caregivers. Behavioral symptoms are complex in nature and require that caregivers understand their presumed causes and intervene appropriately using validated caregiving techniques. Unfortunately, confusing terminology hampers improvement in management techniques. This review attempts to clarify terminology and specifically the behavioral symptoms "agitation" and "resistiveness to care" that require different management techniques. Several conceptual frameworks for behavioral symptoms of dementia are presented. These frameworks include behavioral models, a psychiatric model, and a comprehensive model that combines both behavioral and psychiatric strategies. Using precise terminology consistently and providing care based on a conceptual framework will facilitate the education of caregivers in appropriate techniques for management of behavioral symptoms of dementias. PMID- 14528042 TI - Influence of regular proprioceptive and bioenergetic physical activities on balance control in elderly women. AB - BACKGROUND: Balance disorders increase considerably with age due to a decrease in posture regulation quality, and are accompanied by a higher risk of falling. Conversely, physical activities have been shown to improve the quality of postural control in elderly individuals and decrease the number of falls. The aim of this study was to evaluate the impact of two types of exercise on the visual afferent and on the different parameters of static balance regulation. METHODS: Static postural control was evaluated in 44 healthy women aged over 60 years. Among them, 15 regularly practiced proprioceptive physical activities (Group I), 12 regularly practiced bioenergetic physical activities (Group II), and 18 controls walked on a regular basis (Group III). RESULTS: Group I participants displayed lower sway path and area values, whereas Group III participants displayed the highest, both in eyes-open and eyes-closed conditions. Group II participants displayed intermediate values, close to those of Group I in the eyes open condition and those of Group III in the eyes-closed condition. Visual afferent contribution was more pronounced for Group II and III participants than for Group I participants. CONCLUSIONS: Proprioceptive exercise appears to have the best impact on balance regulation and precision. Besides, even if bioenergetic activity improves postural control in simple postural tasks, more difficult postural tasks show that this type of activity does not develop a neurosensorial proprioceptive input threshold as well, probably on account of the higher contribution of visual afferent. PMID- 14528043 TI - Angiotensin I-converting enzyme gene polymorphism in two ethnic groups living in Brazil's southern region: association with age. AB - BACKGROUND: Several studies have been published on the association between ACE (angiotensin I-converting enzyme) polymorphism and longevity. However, the results are controversial. METHODS: We analyzed the association between ACE polymorphism and age in two different ethnic populations: a population originating from different European countries (Gaucha Population, GP) and a population originating from Japan (JP). Both populations live in Brazil's southern region. RESULTS: We determined the ACE genotype in 834 persons aged 10 104 years. The ACE genotype and allele frequencies differed between the two populations, with the D allele being more frequent in GP than JP. We found an association between the DD genotype and D allele and age in the GP group only. The ACE polymorphism-age association occurred at age >60 years in the GP population with decreasing II frequency. CONCLUSION: We cannot dismiss the possibility of the association between ACE polymorphism and age involving linkage disequilibrium, since the nature of this phenomenon is still controversial. From our studies, it appears that there is a correlation between age, ethnicity, and ACE polymorphism. More of such studies are warranted, as further investigation in this area will have a high clinical relevance to cardiovascular disease and aging research. PMID- 14528044 TI - Prevalence of multivessel coronary artery disease in patients with diabetes mellitus plus hypothyroidism, in patients with diabetes mellitus without hypothyroidism, and in patients with no diabetes mellitus or hypothyroidism. AB - BACKGROUND: We report the prevalence of significant multivessel coronary artery disease (CAD) (>50% stenosis) in patients with diabetes plus hypothyroidism, in patients with diabetes without hypothyroidism, and in patients without diabetes or hypothyroidism who had significant CAD documented by coronary angiography. METHODS: We performed a retrospective analysis in 100 patients selected randomly with significant CAD documented by coronary angiography to investigate the prevalence of significant multivessel CAD in patients with diabetes, in patients with hypothyroidism, and in patients without diabetes or hypothyroidism. RESULTS: Significant 3-4-vessel CAD was present in 9 of 10 patients (90%), mean age 73 +/- 11 years, with diabetes plus hypothyroidism, in 10 of 25 patients (40%), mean age 63 +/- 9 years, with diabetes without hypothyroidism, and in 10 of 65 patients (15%), mean age 68 +/- 12 years, without diabetes or hypothyroidism (p <.001 comparing diabetes plus hypothyroidism with no diabetes or hypothyroidism; p <.01 comparing diabetes plus hypothyroidism with diabetes without hypothyroidism; and p <.02 comparing diabetes without hypothyroidism with no diabetes or hypothyroidism). Significant 2-4-vessel CAD was present in 10 of 10 patients (100%) with diabetes plus hypothyroidism, in 20 of 25 patients (80%) with diabetes without hypothyroidism, and in 37 of 65 patients (57%) with no diabetes or hypothyroidism (p <.01 comparing diabetes plus hypothyroidism with no diabetes or hypothyroidism; and p <.05 comparing diabetes without hypothyroidism with no diabetes or hypothyroidism). CONCLUSIONS: In patients with diabetes mellitus plus hypothyroidism, the prevalence of 3-vessel or 4-vessel CAD was significantly higher than in patients with diabetes mellitus without hypothyroidism or in patients with no diabetes mellitus or hypothyroidism. In patients with diabetes mellitus without hypothyroidism, the prevalence of 3-vessel or 4-vessel CAD was significantly higher than in patients without diabetes mellitus or hypothyroidism. PMID- 14528045 TI - Association of abnormal serum lipids in elderly persons with atherosclerotic vascular disease and dementia, atherosclerotic vascular disease without dementia, dementia without atherosclerotic vascular disease, and no dementia or atherosclerotic vascular disease. AB - BACKGROUND: Observational data have shown that the use of statins was associated with a lower prevalence of vascular dementia and of Alzheimer's disease. METHODS: We investigated in an academic nursing home the association of dyslipidemia with atherosclerotic vascular disease (group 1), atherosclerotic vascular disease without dementia (group 2), dementia without atherosclerotic vascular disease (group 3), and no dementia or atherosclerotic vascular disease (group 4). RESULTS: Increased serum low-density lipoprotein (LDL) cholesterol was present in 36 of 50 group 1 patients (72%), in 34 of 50 group 2 patients (68%), in 34 of 50 group 3 patients (68%), and in 18 of 50 group 4 patients (36%) (p =.0003 for 1 versus 4; p = 0.001 for 2 versus 4 and 3 versus 4). Decreased serum high-density lipoprotein (HDL) cholesterol was present in 34 of 50 group 1 patients (68%), in 35 of 50 group 2 patients (70%), in 32 of 50 group 3 patients (64%), and in 16 of 50 group 4 patients (32%) (p =.0003 for 1 versus 4; p =.0001 for 2 versus 4; p =.001 for 3 versus 4). Hypertriglyceridemia was not significantly different in the 4 groups. CONCLUSIONS: The prevalence of increased serum LDL cholesterol and decreased serum HDL cholesterol were significantly higher in elderly patients with atherosclerotic vascular disease plus dementia, atherosclerotic vascular disease without dementia, and dementia without atherosclerotic vascular disease than in patients with no dementia or atherosclerotic vascular disease. PMID- 14528046 TI - Older women with osteoporosis have increased postural sway and weaker quadriceps strength than counterparts with normal bone mass: overlooked determinants of fracture risk? AB - BACKGROUND: Fracture risk is determined by both bone health and fall risk. Evidence suggests that older women with osteoporosis may have a greater risk of falling compared with their age-matched counterparts without osteoporosis ( 1). To determine whether fall risk screening should be a routine part of medical assessment in older women with osteoporosis, a comparison of fall risk between those with osteoporosis and healthy age-matched counterparts is needed. The purpose of this study was to compare 3 established fall risk factors between these 2 groups of women. METHODS: 42 women between the ages of 64 and 75 years old participated in this study. 21 women with osteoporosis were matched by age and current physical activity level to 21 women without osteoporosis. The performance on 3 fall risk factors (quadriceps strength, balance, and functional mobility) was compared between the 2 groups using multivariate analysis of variance. The level of significance was set at p T, G590-->A and G857-->A). The NAT2 phenotype was evaluated with dapsone as a test substrate using high-pressure liquid chromatography. Statistical analysis was performed using the chi(2) test. Phenotype and genotype were distributed as follows: (1) of the healthy subjects, 60% were rapid acetylators (RA) and 40% were slow acetylators (SA); 10% of the RA and 15% of the SA were homozygous, 50% of the RA and 25% of the SA were heterozygous; (2) of the patients, 55% were RA, 40% were SA and 5% were intermediate acetylators (IA); 10% of the RA and 10% of the SA were homozygous, 45% of the RA and 35% of the SA were heterozygous. No significant statistical difference was found between the two groups for genotypes (p = 0.75) or phenotypes (p = 0.60). The phenotyping and genotyping results of healthy subjects were comparable to those found in previous studies. The absence of a significant statistical difference between healthy subjects and atopic dermatitis patients is in contrast to the results of previous studies. Some authors considered that allergic patients are mostly SAs. This could be explained by the fact that we only considered patients suffering from atopic dermatitis whereas, in other studies, patients suffered from different (one or several associated) allergic diseases. NAT2 polymorphism does not differ between patients suffering from atopic dermatitis and healthy subjects. The importance attributed to the SA status, which was previously considered a predisposing factor for allergic diseases such as atopic dermatitis, should be reviewed. PMID- 14528064 TI - Role of isopropyl myristate, isopropyl alcohol and a combination of both in hydrocortisone permeation across the human stratum corneum. AB - The influence of isopropyl myristate (IPM), isopropyl alcohol (IPA) and a combination of both was studied in view of hydrocortisone (HC) permeation across the human stratum corneum (SC). IPM, IPA and their combination were incorporated into water-containing hydrophilic ointment (WHS), and the resulting effects on HC permeation and on HC accumulation in human SC were investigated as well as the influence of these substances on the microstructure of the SC. Differential scanning calorimetry as well as wide- and small-angle X-ray diffraction show that IPM incorporation into SC results in densely packed bilayer lipids and a loss of order of the corneocyte-bonded lipids. Both effects result in a decreased diffusion coefficient of HC in SC and thus in a decreased permeation rate compared to that of HC from WHS. On the other hand, IPA fluidizes and disrupts the bilayer structure of the intercellular lipids. These effects, concomitant with an increased amount of dissolved HC within the ointment, increase the permeation rate of HC across SC. The combination of both ingredients effects a stronger fluidization and disruption of intercellular lipids than with IPA alone. Therefore, the permeation rate of HC across SC is higher than with IPA alone. Consequently, the IPM and IPA combination acts synergistically on the microstructure of SC. PMID- 14528065 TI - Epinastine inhibits eosinophil chemotaxis and adhesion molecules in atopic dermatitis. AB - PURPOSE: To investigate the effects of epinastine on eosinophil chemotaxis and changes in eosinophil adhesion molecules induced by epinastine and three other antiallergic agents, using eosinophils of atopic dermatitis (AD) patients. RESULTS: Epinastine reduced eosinophil chemotaxis toward eotaxin when the eosinophils had been prestimulated with interleukin (IL)-5, but given alone it did not alter eosinophil chemotaxis toward IL-5. CD11b expression was inhibited when peripheral blood was prestimulated with IL-5, but eosinophil adhesion molecule expression was not altered. CONCLUSIONS: Epinastine suppresses allergic inflammation not only through its strong antihistamine and antimediator effects, but also by inhibiting eosinophilic chemotaxis and the expression of adhesion molecules involved in chemotaxis, especially CD11b. PMID- 14528066 TI - Cutaneous hand and foot toxicity associated with cancer chemotherapy. PMID- 14528067 TI - Waldeyer's ring lymphomas: treatment results and prognostic factors. AB - Optimal management of patients with localized Waldeyer's ring (WR) lymphoma remains controversial due to the lack of randomized studies and heterogenous grouping of most reported series. In this retrospective study, we have evaluated the possible prognostic factors and treatment outcome of WR non-Hodgkin's lymphoma. Between December 1993 and February 2000, 32 patients with WR lymphoma, stage I (11 patients) and stage II (21 patients) were treated. There were 17 male patients and 15 female patients with a median age of 47 years. The distribution among different anatomical sites were as follows: tonsils in 16 (50%), nasopharynx in 10 (31%), base of tongue in 6 (19%). According to Working Formulation, 10 had high-grade, 17 intermediate grade, 3 low-grade, and 2 had unclassified lymphomas. Combined chemotherapy and radiotherapy was the primary modality of therapy for intermediate or high-grade lymphoma. Radiotherapy alone was employed only in low-grade WR lymphomas. Chemotherapy was median 6 courses of CHOP (cyclophosphamide, doxorubicin (Adriamycin), vincristine, and prednisolone) in 26 patients and CEOP (cyclophosphamide, doxorubicin, etoposide, and prednisone). Radiotherapy volume was involved field and the median dose was 40 Gy. Median follow-up is 40 months (ranged from 6-82 months). Overall survival and disease-free survival (DFS) rates at 3 years are 100% and 92%, respectively. Two patients developed recurrence, both salvaged with further chemotherapy. Only one patient died because of other reasons. International Prognostic Index score (2) is found to be an important prognostic factor for DFS. The other significant prognostic factors for DFS are performance status and serum levels of alkaline phosphatase and lactate dehydrogenase. Our results suggest that combined chemotherapy and involved field radiotherapy is appropriate treatment for stage I II WR lymphoma. International Prognostic Index is the strongest predictor for DFS. PMID- 14528068 TI - Weekly carboplatin and paclitaxel in elderly non-small-cell lung cancer patients (>or=65 years of age): a phase II North Central Cancer Treatment Group study. AB - The purpose of this study was to determine the tumor response rate and toxicity profile of low-dose weekly carboplatin and paclitaxel in advanced non-small-cell lung cancer (SCLC) patients 65 or more years of age. Forty-nine patients 65 years of age or more with advanced non-SCLC with a median age of 73 years (range: 65 85) and an Eastern Cooperative Oncology Group performance status of 0, 1, or 2 in 31%, 47%, and 22% of patients, respectively, were treated and evaluated. Patients received carboplatin (AUC = 2) and paclitaxel 50 mg/m2 on days 1, 8, and 15 of a 4-week cycle. The overall confirmed tumor response rate was 14% (95% CI: 4.7%, 32.5%) with no complete responses. The 1-year survival rate was 31% (95% CI: 20%, 48%). There was one treatment-related death, and there were two grade IV allergic reactions to chemotherapy. No other grade IV or V treatment-related toxicities were observed. There were only three episodes of grade III myelosuppression. Low dose weekly carboplatin and paclitaxel, as prescribed in this trial, provides modest activity in the treatment of advanced non-SCLC patients 65 or more years of age. However, the relatively mild toxicity profile observed in this trial suggests that this regimen might remain an option for patients at increased risk for myelosuppression or with a poor performance status. PMID- 14528069 TI - Hematopoietic protection by dexamethasone or granulocyte-macrophage colony stimulating factor (GM-CSF) in patients treated with carboplatin and ifosfamide. AB - Based on preclinical studies, the authors undertook a pilot study to determine the hematologic and biologic effects of pretreatment with dexamethasone (Dex) or granulocyte-macrophage colony-stimulating factor (GM-CSF) in patients receiving carboplatin and ifosfamide. Patients (n = 28) with metastatic solid tumors were randomized to receive pretreatment with Dex or GM-CSF or no pretreatment prior to courses 1 or 2 of carboplatin and ifosfamide. No alteration in dose of chemotherapy was allowed between course 1 and 2. Alterations of hematologic and nonhematologic toxicity and selected biologic parameters were compared between courses 1 and 2. Patients without any pretreatment demonstrated worsening hematologic toxicity in course 2 compared to course 1. In contrast, Dex pretreatment reduced hematopoietic toxicity and improved the absolute granulocyte count (AGC) and platelet count recovery times. For example, course 1 versus course 2 (with Dex pretreatment): AGC nadir (mm3) 153 versus 549 (p = 0.07), days AGC <500/mm3 7.8 versus 4.0 (p = 0.10), days to AGC recovery >1,500/mm3, 26 versus 22 (p = 0.034). Overall comparison between all five cohorts by analyses of variance demonstrated that intervention with Dex improved multiple hematopoietic toxicities, including AGC nadir (p = 0.015), and recovery times to AGC >1,500/mm3 (p = 0.07) and platelet count to >100,000/mm3 (p = 0.05). GM-CSF pretreatment did not worsen hematopoietic parameters after course 2 compared to course 1. Expected biologic effects of Dex and GM-CSF treatment were observed. Patients demonstrated an overall response rate of 32%, 1 complete response, and 8 partial responses. In patients with cancer, pretreatment with Dex or GM-CSF may significantly decrease the hematopoietic toxicity of chemotherapeutic agents. PMID- 14528071 TI - Algorithm for the diagnosis of HER-2/neu status in breast-infiltrating carcinomas. AB - Determination of HER-2/neu overexpression/amplification is becoming increasingly important. The aim of this study is to elaborate an algorithm for the diagnosis of Her-2/neu status in breast-infiltrating carcinomas. Three hundred five breast infiltrating carcinomas were selected to determine HER-2/neu overexpression by immunohistochemistry with two different methods: the monoclonal antibody CB11 and the HercepTest. Fluorescence in situ hybridization (FISH) was performed in a subgroup of those cases. Time-consuming and reagent costs were calculated for each of the procedures. HER-2/neu overexpression was found in 16% and 33% of the tumors with the monoclonal antibody (mAb) CB11 and with the HercepTest, respectively. There were 50 cases with immunohistochemical discordant results; most of them were HercepTest score 2+/mAb CB11 negative (37/50). Of those cases, only 27% presented gene amplification. The algorithm consisted of testing all the specimens with the mAb CB11 for selection of positive cases, negative cases are confirmed with the HercepTest, and FISH is performed only in those cases with immunohistochemical discordant results. The algorithm rate of HER-2/neu positivity in our series was 22%. Time and costs are reduced by 60% and 41%, respectively, compared to FISH. The use of the algorithm is feasible, accurate, and cost-effective in relation to FISH. PMID- 14528070 TI - Retrospective analysis of safety and efficacy of low-dose docetaxel 60 mg/m2 in advanced non-small cell lung cancer patients previously treated with platinum based chemotherapy. AB - The efficacy and toxicity of low-dose docetaxel (60 mg/m2) were evaluated in patients with relapsed non-small-cell lung cancer (NSCLC) after platinum containing chemotherapy. Docetaxel 60 mg/m2 was infused during 1 hour with no routine premedication, with courses repeated at 3-week intervals. Twenty-seven patients were analyzed retrospectively. The median age was 56 years (range, 32 72); 22 patients (81.5%) had adenocarcinoma, 26 (96.3%) had stage IV disease, and 23 (85.2%) were Eastern Cooperative Oncology Group performance status 0 to 1. Five patients (18.5%) had a partial response. Median progression-free survival time for all patients was 1.9 months, and median survival time was 9.4 months. The predominant toxicity was neutropenia, which was grade III or IV in 63% of patients. No neutropenic fever was observed. Other hematologic toxicities were mild (all grade II). Thus, low-dose docetaxel (60 mg/m2) yielded a response rate comparable to that achieved with moderate- to high-dose docetaxel (75-100 mg/m2) as second-line chemotherapy in platinum-pretreated NSCLC, and had less toxicity. Further investigation of the optimal docetaxel dose as second-line chemotherapy in NSCLC is warranted. PMID- 14528072 TI - Angiosarcoma of the breast occurring soon after lumpectomy and breast irradiation for infiltrating ductal carcinoma: a case report. AB - Angiosarcoma of the breast is a rare second malignancy after breast irradiation for carcinoma. As with most radiotherapy-associated malignancies, the usual latent period after radiotherapy is many years. The authors present a case of angiosarcoma appearing in the breast 14 months after breast irradiation after lumpectomy and axillary node dissection for infiltrating ductal carcinoma. PMID- 14528073 TI - Benign metastasizing leiomyomatosis: case report and review. AB - The authors report an interesting case of a minimally symptomatic 23-year-old African American woman who was found to have extensive diffuse reticulonodular opacities of the lungs on a routine chest radiograph. She had a hysterectomy 5 years previously for multiple leiomyomas of the uterus. She had no history of any prior exposure to dusts or toxins. Collagen vasculitides and bacterial, mycobacterial, and fungal infectious causes were excluded through standard testing, and a bronchoscopic lung biopsy was nonspecific. An open lung biopsy revealed multiple nodules of proliferating smooth muscle cells intermixed with irregular areas of epithelial-lined spaces. Histologically, the muscle cells appear benign with a very low mitotic rate, and the pathologic findings were consistent with benign metastasizing leiomyomatosis (BML). Staining for estrogen and progesterone receptors, actin, and c-kit were performed. This case and the review of the medical literature support the concept that BML originates from an antecedent leiomyoma of the uterus in virtually all cases with rare exceptions. It appears that tumor metastasizes to lungs or other extrauterine tissues via hematogenous spread. However, the origin of the tumor remains controversial. BML is a rare entity, with only a handful of reports in the medical literature. The authors report an interesting case of BML in a 23-year-old patient who, to their knowledge, is the youngest such patient described and who, at 13 years, has the longest period of clinical follow-up. In this article, the authors review the pathogenesis, cytogenetics, histologic markers, and management options of this rare entity. PMID- 14528075 TI - Small bowel adenocarcinoma in crohn disease patient complicated by microangiopathic hemolytic anemia. PMID- 14528074 TI - Docetaxel as neoadjuvant chemotherapy in patients with advanced cervical carcinoma. AB - The purpose of this study was to evaluate the efficacy and toxicity of docetaxel as single-agent neoadjuvant chemotherapy in locoregionally advanced cervical carcinoma. Between April 1998 and August 2000, 38 untreated patients with International Federation of Gynecology and Obstetrics stages IIB to IVA were entered onto this study. The median age was 44 years (range: 25-66 years). Stages: IIB 22 patients, IIIB 15 patients, and IVA 1 pt. Treatment consisted of docetaxel 100 mg/m2 IV infusion during 1 hour. Standard premedication with dexamethasone, diphenhydramine, and ranitidine was used. Cycles were repeated every 3 weeks for three courses, followed by radical surgery when it was judged appropriate, or definitive radiotherapy. Both staging and response assessment were performed by a multidisciplinary team. 106 cycles of therapy were administered; all patients were evaluable for TX, whereas 35 were evaluable for response (3 patients refused further treatment after the first cycle of therapy). Complete response (CR): 1 patient (3%); partial response: 11 patients (31%), for an overall objective response rate of 34% (95% CI: 15-53%); no change (NC): 16 patients (46%); and progressive disease: 7 patients (20%). Six patients (17%) underwent surgery and a pathologic CR was confirmed in 1 of them. The median time to treatment failure and the median survival have not been reached yet. The limiting toxicity was leukopenia in 25 patients (69%) (G1-G2: 14 patients, G3: 10 patients, and G4: 1 patient). Neutropenia: 28 patients (78%) (G1-G2: 10 patients, G3: 8 and G4: 10). Myalgias: 17 patients (47%) (G1-G2: 15 patients and G3: 2 patients). Emesis: 21 patients (55%) (G1-G2: 19 patients and G3: 2 patients). Alopecia G3: 13 patients (36%); rash cutaneous 26 patients (68%) (G1-G2: 22 patients and G3: 4 patients). There were no hypersensitivity reactions or fluid retention syndrome. The received dose intensity was 91% of that projected. Docetaxel is an active drug against advanced cervical carcinoma with moderate toxicity. Further evaluation in association with other agents is clearly justified. PMID- 14528076 TI - Angiosarcoma developing in a breast after conservation treatment for breast cancer. PMID- 14528077 TI - Renal medullary carcinoma: case report and review of the literature. AB - Renal medullary carcinoma is a recently recognized epithelial malignant tumor arising in the renal parenchyma. The tumor is almost exclusive to young black patients with the sickle cell trait. Most patients present with metastatic disease and have a poor prognosis. An Hispanic woman with renal medullary carcinoma who initially responded to chemotherapy with methotrexate, vinblastine, doxorubicin, and cisplatin and survived for 12 months is presented. The clinical, histologic, and radiologic features of this tumor are described, and chemotherapeutic regimens used in this disease are detailed. Treatment modalities have proved largely unsuccessful in the setting of advanced disease. Given the shared demographic, clinical, and radiographic features of these patients, awareness and early diagnosis may prove essential in improving survival. PMID- 14528078 TI - Aerosolized granulocyte macrophage colony-stimulating factor (GM-CSF) therapy in metastatic cancer. AB - A recent phase I study of aerosolized granulocyte macrophage colony-stimulating factor (GM-CSF) in patients with malignant metastases to the lungs demonstrated excellent tolerance and possible efficacy. This therapy was offered to other patients who refused "standard" treatment or when no effective therapy was available. Forty-five patients were treated; 40 had pulmonary metastases. Aerosolized GM-CSF (250 microg/dose) was administered twice a day using a 1 week on, 1 week off schedule. The mean interval between diagnosis and therapy was 32 months. Twenty-four patients had disease stabilization or partial regression. The mean duration of benefit was 10 months. This benefit was noted in 8 of 13 with a sarcoma, 6 of 14 with melanoma, and 5 of 12 with renal cell carcinoma. Eighteen patients reported mostly self-limiting toxicities. The frequency of certain melanoma-specific T lymphocytes in 1 patient with stable disease was found to have increased 10-fold after therapy. Aerosolized GM-CSF appears to have limited but promising efficacy in treatment of pulmonary metastatic disease. In one patient, we have evidence of upregulation of melanoma-specific cytotoxic T-cells. Further study is warranted to understand the impact of this therapy on the natural history of metastatic cancer. PMID- 14528079 TI - Tumor dimension and prognosis in surgically treated lung cancer: for intentional limited resection. AB - Tumors with a maximum dimension of 3 cm are categorized as T1, whereas those greater than 3 cm are T2 by TNM classification. Some physicians suggest that early-stage peripheral lung cancer should have a maximum tumor diameter of 2 cm and that limited surgery (segmentectomy without lymph node dissection) is acceptable for the patients. In this study, the relationship between the tumor dimension and prognosis was analyzed in 207 patients with surgically treated primary non-small-cell lung cancer (SCLC). The 5-year survival rate of those with tumors 3 cm or less and without lymph node (LN) metastases was 86%, which was significantly higher than that of those with tumors more than 3 cm and without hilar and mediastinal LN metastases (65%) (p < 0.05). However, 33% of the patients with tumors 3 cm or less had LN metastases, and the 5-year survival rate did not differ between those with tumors 3 cm or less (60%) and those with tumors more than 3 cm (54%). Twenty-eight percent of patients with tumors 2 cm or less had LN metastases, and the 5-year survival rate of the patients with tumors 2 cm or less was 62%. The 5-year survival rate of those with tumors 2 cm or less and without LN metastases was 88%. Forty-six patients with tumors 2 cm or less included 5 cases with an intrapulmonary metastasis in the same lobe (11%). In conclusion, a size of 3 cm is an appropriate boundary as the T factor. Because those with tumors 2 cm or less have a relatively high percentage of LN metastases, intraoperative frozen sections of LN should be considered for those undergoing limited surgery for primary non-SCLCs 2 cm or less. Intrapulmonary metastases also should be considered for those undergoing limited surgery even if the maximum dimension of the primary tumor is less than 2 cm. PMID- 14528080 TI - Favorable outcome of preoperative low dose chemoradiotherapy against muscle invasive bladder cancer. AB - To investigate the feasibility of preoperative low-dose chemoradiotherapy, 50 patients with localized muscle-invasive bladder cancer (T2-T4) were treated with concurrent cisplatin (100 mg/body x 2 courses) and pelvic irradiation (40 Gy). Among 20 patients (40%) who achieved clinical complete regression, 11 with solitary tumor underwent partial cystectomy because of advanced age, poor condition of the patients, or a reluctance to have radical surgery. Radical cystectomy was carried out in the remaining 39 cases (complete regression 9, partial regression 30). Pathologic T0 response (no residual tumor) was achieved in 18 (36%) of all the cases. Median follow-up was 19 months (range 2-59 months). Estimated 3-year disease-free survival was 75% for all patients and 100% for T0 responders. Local recurrence (2 patients) or distant metastasis (6 patients) developed in 8 of 32 patients with pathologic persistent tumor. All of the 11 patients (9 with T0 response and 2 with a small residual tumor nest) who underwent partial cystectomy were recurrence free for observation periods up to 59 months. Low-dose chemoradiotherapy is as effective as other neoadjuvant protocols and is applicable to high-risk patients. The bladder can be preserved by partial cystectomy in selected cases with clinical complete regression. PMID- 14528081 TI - Endocavitary radiation therapy for rectal adenocarcinoma: 10-year results. AB - Local control, survival, and toxicity in patients treated with endocavitary radiation therapy for rectal cancer were evaluated. Thirty-five patients received a total of 20 to 155 Gy in 1 to 5 fractions with 50 kV x-rays through a treatment proctoscope. Twenty-nine of the 35 patients were treated with curative intent. Median follow-up was 102 months. Local control was achieved in 23 of the 29 patients treated curatively and in 3 of the 6 treated palliatively. Local control for patients treated curatively was 76% at 10 years. No local failures occurred after 21 months. For patients treated curatively, survival was 65% at 5 years and 42% at 10 years. Toxicity within 90 days after treatment was observed in 77% of the patients. Toxicity occurring more than 90 days after treatment was observed in 80%, but only 1 patient needed a colostomy, which was for a perforation after the biopsy of a benign ulcer. In conclusion, radiation therapy resulted in a local control rate of 76% at 10 years in curatively treated patients. Although most patients experience toxicity from this treatment, loss of sphincter function is rare. PMID- 14528082 TI - Marriage and fertility in long-term survivors of high grade osteosarcoma. AB - One hundred fifteen patients with high grade osteosarcoma were treated at our hospital between 1976 and 1995. Of these, 45 patients still surviving and disease free for more than 5 years since the conclusion of treatment were enrolled as the subjects. The marriage proportion (number of married persons/number of all persons) and fertility proportion (number of persons having an offspring/number of married persons) of these 45 patients were investigated. The marriage proportion of male patients was 20.8% (5/24), whereas that of female patients was 76.2% (16/21). The marriage proportion in male patients was dramatically lower than that in female patients (p = 0.0003). The marriage proportion in male patients was significantly lower than that in brothers of the patients (p = 0.0223). In female patients, the treatment of osteosarcoma had no influence on marriage, whereas in male patients it had a great influence on marriage. The fertility proportion in married patients was 76.2% (16/21) and that in siblings of the patients was 81.0% (17/21). A total of 18 offspring were born to married patients. These offspring had no birth defects or congenital anomalies. The chemotherapy used in the treatment of osteosarcoma had little influence on the fertility of patients or the health of their offspring. PMID- 14528083 TI - Granulosa cell tumor of the ovary: retrospective analysis of 45 cases. AB - Adult granulosa cell tumors of the ovary are rare neoplasms, accounting for less than 5% of all ovarian malignancies. In addition to the tumor stage, residual disease, patient age, tumor size, extent of surgery, and also some histologic factors have been reported to be of prognostic importance. Tumor registries were screened for all patients treated between 1979 and 1998 for ovarian tumors at the University of Istanbul. There were 952 ovarian carcinomas, of which 47 were granulosa cell tumors. All charts were reviewed, and the clinical data were extracted. Prognostic factors and treatment results were evaluated retrospectively. The median follow-up was 84 (range: 6-141 months) months. According to univariate analysis, there were only two significant factors for overall survival (OS): stage and presence of residual disease. The OS of the 23 patients with early stage (mean, 122 months; median, unreached) was significantly (p = 0.0001) better than the OS of the 22 patients with advanced stage (mean, 34 months; median, 21 months). A significant difference (p = 0.0004) in OS was also observed between patients with residual (mean, 42 months; median, 21 months) and nonresidual (mean, 108 months; median, unreached) disease. In a multivariate analysis, only stage remained statistically significant (p = 0.0001). The overall 5-year survival rate was 55% and median survival after recurrence was 21 months. Despite the small number of patients, the study showed that stage and macroscopic residual disease are significant prognostic factors. The benefit of chemotherapy and radiotherapy remains controversial. PMID- 14528084 TI - Report from the Radiation Oncology Committee of the Southwest Oncology Group (SWOG): Research Objectives Workshop 2003. AB - To achieve the ultimate goal of cancer treatment, which is 100% cancer control with negligible toxicity, the therapeutic window must be enlarged, allowing for higher doses of beneficial treatments with reduced toxicity. The advent of image- and metabolism-guided therapy offers the best opportunity to date for combining modern radiation targeting and imaging techniques. Indeed, for the first time, it is reasonable to locally target metastatic disease with the goal of sterilization. Combining these focal radiation techniques with novel targeted antiproliferative agents and full-dose classic cytotoxic chemotherapy will become more effective as we learn to use these compounds in a less systemically toxic manner and as radiation fields become more defined. In addition, increasing numbers of biologic modifiers of normal tissue response are becoming available, and they suggest great promise for decreasing the normal tissue toxicity resulting from both radiation and chemotherapy treatments. Thus, radiation metastectomy for gross metastases, used together with systemic control of micrometastatic disease, may yield improved survival rates. This hypothesis is ready for testing in cancers of the breast, prostate, colon, and in sarcomas. Enlarging the therapeutic window is a major goal that would allow for an increasingly favorable therapeutic gain. PMID- 14528085 TI - Intermittent androgen deprivation for patients with recurrent/metastatic prostate cancer. AB - This study was designed to assess the duration of response to intermittent androgen deprivation therapy (IAD) in patients with recurrent and/or metastatic prostate cancer. Between January 1993 and March 2000, 74 patients with recurrent and/or metastatic prostate cancer had IAD with either luteinizing hormone releasing hormone agonist (LHRH) or an LHRH with an oral antiandrogen. Forty-one patients were treated for an increasing prostate-specific antigen (PSA) level after primary local treatment. Of the remaining 33 patients, 17 patients were treated for metastases (9 for bone metastases, 8 for lymph nodes metastases, and 16 for local recurrence). Patients who had undergone IAD completed between 1 and 6 cycles. A cycle was defined as the period during which the patient was actively taking the hormone medication. Seventy-four patients completed the first cycle, 49 completed the second cycle, and 23 completed the third cycle. The pattern of PSA changes with each cycle, the length of each cycle, and the time interval between successive cycles were studied. The time to progression (defined as an increasing PSA level on two consecutive measurements or radiologic evidence of progression of disease while the patient was on androgen deprivation) was also studied. The median PSA before the IAD was 11.4 ng/mL (range 0.12-378). The median PSA nadir at the end of each cycle increased progressively (0.1 ng/mL after the first cycle to 3.3 ng/mL after the fifth cycle). The time interval between the cycles progressively decreased from 9.5 months between the first and second cycles to 6 months between the third and fourth cycles. The 4-year actuarial androgen-independent free survival was 71%. For the subgroups of patients treated for biochemical failure, locoregional recurrence, and bone metastases, the 4-year actuarial progression-free survival rates were 80%, 67%, and 45% respectively (P = 0.018). The median time of 18 months to progression in patients with bone metastases is similar to that reported with continuous hormonal therapy. In patients with biochemical failure, the median time to progression (more than 5 years) suggests that the IAD approach may be a viable option for this group of patients. PMID- 14528087 TI - Impact of intraoperative edema during transperineal permanent prostate brachytherapy on computer-optimized and preimplant planning techniques. AB - The purpose of this study was to prospectively evaluate intraoperative prostatic edema during prostate brachytherapy with real-time ultrasound imaging and assess its impact upon the postimplant dosimetry of computer-optimized intraoperatively planned patients. Fifty consecutive patients with early-stage favorable risk adenocarcinoma of the prostate underwent transperineal ultrasound-guided I125 brachytherapy. Ultrasound volume studies of the prostate were performed immediately before and after placement of brachytherapy needles in the operating room. Twenty-five patients underwent intraoperative computer-optimized treatment planning using a genetic algorithm. Twenty-five patients underwent preimplant ultrasound studies for preimplant treatment planning. Postimplant dosimetry was performed on computed tomography scans obtained after the implant. Statistical analysis was performed taking into account patient age, preneedle volume, increase in intraoperative edema, use of hormonal therapy, type of isotope, number of needles or seeds used, and seed activity. For the intraoperatively planned patients, a median increase of 30% in intraoperative volume was found for the entire group. No correlation between the extent of intraoperative edema and %D90 (percentage of prescribed dose that covers 90% of the target volume) was found. None of the other analyzed variables correlated with %D90. Patients whose treatment was planned preoperatively experienced a median increase of 18.4% in target volume. A negative correlation between the amount of edema and the %D90 was found to be statistically significant (-0.55, P = 0.0047). All patients who underwent prostate brachytherapy experienced intraoperative prostatic edema. When planned intraoperatively, the amount of edema had no impact on the %D90. This may be because of the ability of intraoperative computer-optimized treatment planning to account for edema related to the procedure. Preplanned patients who encountered a greater degree of intraoperative edema had less %D90 target coverage. PMID- 14528086 TI - Radioresistance in oral squamous cell carcinoma with p53 DNA contact mutation. AB - Reliable variables to predict the radiosensitivity of each tumor have not been identified. Recent studies have demonstrated that specific regions of mutations within the core domain of p53 protein correlate with responses to chemotherapy and radiotherapy in some tumor types. In this study, we evaluated the relationship between specific p53 mutations and radiosensitivity in 49 patients with oral squamous cell carcinomas (SCCs) who underwent preoperative radiotherapy. Exons 5 through 8 of the p53 gene were examined by polymerase chain reaction-single-strand conformation polymorphism and direct sequencing. We detected p53 mutations in 27 (55.1%) cases. DNA contact mutations were detected in 11 (40.7%) of these 27 cases in L3 loop, loop-sheet-helix motif, and zinc binding residues. Tumors containing p53 DNA contact mutations had significantly poorer responses to radiation than the other tumors, although no statistically significant difference between tumors with and without p53 mutations was found. These data indicate that DNA contact mutation of p53 could be a useful marker to predict the radioresistance of oral SCCs. PMID- 14528088 TI - Number and orientation of beams in inversely planned intensity-modulated radiotherapy of the female breast and the parasternal lymph nodes. AB - Intensity-modulated radiotherapy (IMRT) provides better sparing of normal tissue. We evaluated the optimum beam configuration for IMRT based on inverse treatment planning in adjuvant radiotherapy for breast cancer in a case of left-sided tumor. In addition to radiotherapy planning with the conventional technique of tangential wedged 6-MV photon beams and an oblique 15-MeV electron beam, we performed inversely planned IMRT with the step-and-shoot-technique. Dose calculation was carried out using the treatment planning system Virtuos with the inverse optimization module KonRad adapted to it. IMRT plans were generated for 2 to 16 beams. The results were compared with conventional techniques. For a maximum treatment time of 20 minutes, it is shown that IMRT with 12 modulated photon beams and 7 intensity steps is best suited for treatment in the presented case. Compared with a conventional technique with photons combined with electrons, dose conformality and homogeneity of the planning target volume was increased. The mean heart dose was reduced from 9.1 Gy to 6.1 Gy. The volume of heart irradiated with a dose higher than 30 Gy was reduced from 7.6% to 1.9%, and the volume of the left lung from 13.6% to 11.5% as well. Inverse optimization for IMRT with multiple beams is feasible in the adjuvant treatment of breast cancer. Because of the reduction of the high-dose area of a substantial cardiac volume, it is superior to conventional techniques in cases where the parasternal lymph nodes should be integrated into the target volume. Here, a clinical advantage might be detectable. PMID- 14528089 TI - Patient-reported toxicity correlated to dose-volume histograms of the rectum in radiotherapy of the prostate. AB - We studied 73 patients treated with 3-dimensional conformal radiotherapy for prostate cancer to determine whether there is a correlation between dose per volume to either the whole rectum, rectal wall, rectal surface, or anal canal and the development of late rectal complications measured with prostate-specific quality-of-life (QOL) questionnaires. Given doses were 66.6 to 72 Gy. The prostate cancer modules used were the UCLA-Prostate Cancer Index module (UCLA PCI) (5 questions), the Functional Assessment of Cancer Therapy-Prostate module (FACT-P) (1 question), and the European Organization for Research and Treatment of Cancer prostate cancer module (EORTC QLQ-PR25) (4 questions). A Spearman correlation analysis between the total toxicity score and the dose-volume histograms (DVHs) was performed. All statistical tests were 2-sided. Sixty-five (89%) patients returned the questionnaire, and 18 (28%) underwent endoscopy for rectal bleeding. We found that only patients who had had an endoscopy showed a correlation between rectal toxicity and dose per volume, as compared with the other patients who showed none. Correlation between rectal toxicity and dose per volume for all 4 structures was stronger for higher doses. For 70 Gy, all contours, except the anal canal, showed a significant dose-volume correlation. Our results indicate that only in cases of pronounced rectal toxicity is there a dose-volume correlation, especially for doses of 70 Gy or more. DVHs of the whole rectum, wall, or surface, but not the anal canal, are all equivalent in predicting late rectal toxicity. PMID- 14528090 TI - Enhancement of radiosensitivity in head and neck cancer cells by ZD1839 ('IRESSA'), a selective epidermal growth factor receptor tyrosine kinase inhibitor. AB - Overexpression of epidermal growth factor receptor (EGFR) is frequently observed in many solid tumor types, including head and neck squamous cell carcinomas (HNSCC). Recent laboratory experiments have demonstrated that high EGFR levels correlate with increased tumor resistance to radiation. This study investigated the relationship between EGFR expression levels and radiosensitivity in 5 HNSCC cell lines (HSC2, HSC3, HSC4, SCC25, and Ca9-22) and whether treatment with ZD1839 ('Iressa'), a selective EGFR-tyrosine kinase inhibitor (TKI), would improve tumor cell response to radiotherapy. ZD1839 suppressed the growth of HNSCC cell lines in a dose- and time-dependent manner. Radiosensitivity of these HNSCC cell lines, assessed by a clonogenic survival assay, differed greatly and the expression of EGFR varied. EGFR expression levels (EGFR numbers/cell) correlated with increased tumor resistance to radiation (f[x]= 4.54 X, R2 = 0.715; f[x]: EGFR numbers/cell, X: radiosensitivity; D10). Following exposure of the HNSCC cells to 1.0 microM ZD1839 and radiation (0-10 Gy), greater than additive growth inhibitory effects were observed. These results suggest that ZD1839 could enhance tumor radiosensitivity and inhibit tumor growth after radiation, indicating that this combination could have clinical potential in the treatment of patients with head and neck cancer. PMID- 14528091 TI - Referred otalgia in head and neck cancer: a unifying schema. AB - Pain referred to the ear is a commonly encountered clinical event, and the differential diagnoses that must be considered for pain in a normal ear are numerous. For physicians involved in the treatment of patients with referred ear pain, especially those involved in the care of patients with head and neck malignancies, a basic understanding of the mechanisms involved to produce this phenomenon is required. Several sources offer figures outlining the neuroanatomic basis of nonotogenic ear pain. On occasion, there has been omission of various components in this referred otalgia pathway, however. The authors propose a unified schema and outline potential areas of "nervous system error" giving rise to pain in a clinically normal ear. PMID- 14528092 TI - Estimation of tumor volume in cervical cancer by magnetic resonance imaging. AB - In 32 patients with stage 1 or 2 cervical cancer, preoperative magnetic resonance imaging (MRI) images were compared with corresponding linear measurements made on fresh histopathology specimens. Their clinical tumor diameters recorded as a part of Federation of International Gynecologists and Obstetricians (FIGO) staging were also correlated with the MRI-derived tumor volumes. The locations of neoplastic lesions within the cervix and uterus were identified accurately by MRI as verified in subsequent histopathology examinations. The examination under anesthesia (EUA) diameter (tumor size) was not related to the MRI-derived tumor diameter. Pathologic tumor diameter correlated well with the corresponding diameter in T2-weighted MRI. Tumor volume as measured by MRI was an accurate representation of the local extent of the disease and can be used as an objective measure of cervical cancer at the primary site. Substituting MRI-derived volume in place of clinical tumor diameter in the FIGO staging system will help refine its prognostic significance in patients with both operable and nonoperable cervical cancer. PMID- 14528093 TI - Influence of oxygen on the radiosensitivity of human glioma cell lines. AB - We have investigated the influence of hypoxia on the radiosensitivity of 4 early passage tumor cell lines that were established from malignant glioma patients at our Institute. These cell lines were M006, M059J (a highly radiosensitive line), M059K (a radioresistant line derived from the same biopsy as M059J), and M010b. The GM637 human fibroblast cell line was used as a normal control. The oxygen enhancement ratios (OERs) for these cell lines, determined using a clonogenic survival assay, were approximately 3.6 (GM637), approximately 3.7 (M006), approximately 2.5 (M010b), approximately 2.1 (M059K), and approximately 3.5 (M059J). The broad range of OERs for these glioma lines was not related to cellular glutathione levels or to differences in intrinsic cellular radiosensitivity. Because studies with rodent cell lines indicate that defects in certain DNA repair genes, including ERCC1, can greatly influence cellular OERs, and because several such repair genes, including ERCC1, localize to a region of chromosome 19q that is close to a common deletion in human glioma, we reasoned that such deletions might contribute to the diverse OERs of these tumor cell lines. However, measurements of ERCC1 protein levels using immunofluorescence staining or Western blotting, of ERCC1 mRNA levels using Northern blotting, and of functional nucleotide excision repair capability using the UV/adenovirus reactivation assay, failed to indicate any deficit in these activities. Thus, although the effect of hypoxia on the radiosensitivity of different human glioma cell lines can vary widely, the mechanism of this effect remains unknown. The potential implications of this finding for radiation therapy, and especially for hypoxia imaging-guided intensity-modulated radiation therapy (IMRT) treatment planning, are discussed. PMID- 14528094 TI - Extracapsular radiation dose distribution after permanent prostate brachytherapy. AB - The physical extent of the radiation therapy dose delivered to the periprostatic region was evaluated in 26 consecutive patients with low-risk prostate cancer (prostate-specific antigen or=5.0 mm for all slices evaluated. The utilization of preplanning periprostatic margins is strongly correlated with the administration of prescription radiation doses to the periprostatic region. The extent of the postimplant periprostatic margin (6.5 +/- 1.8 mm) satisfies the preplanning margin criterion of >5 mm. PMID- 14528095 TI - The challenge and cost of patient safety. PMID- 14528096 TI - Learning from failure. PMID- 14528097 TI - Extramedullary leukemia in children with newly diagnosed acute myeloid leukemia: a report from the Children's Cancer Group. AB - OBJECTIVES: To describe features of patients with acute myeloid leukemia presenting with extramedullary leukemic tumors (EML). METHODS: Among 1,832 patients entered on Children's Cancer Group's chemotherapy trials with acute myeloid leukemia, 199 patients had EML, defined as any leukemic collection outside the bone marrow cavity. Three patient groups were denoted: group 1 (n=109) with EML involving skin (with or without other sites of EML), group 2 (n=90) with EML in sites other than skin, and group 3 (n=1,633) without EML. RESULTS: The incidence of EML was 10.9%. Group 1 patients tended to be younger, had higher white blood cell counts, were more often CNS positive, had FAB M4 or M5 subtypes, and possessed more abnormalities of chromosome 11 than group 3 patients. Group 2 patients were younger, more often had the FAB M2 subtype, and had a higher incidence of t(8;21)(q22;q22) abnormality than group 3, but had similar white blood cell counts and incidence of CNS positivity at diagnosis. For group 1 the 5-year event-free survival was 26%, significantly worse than for group 3 at 29%. Event-free survival was better for group 2 patients (5-year estimate 46%), which remained a favorable prognostic factor by multivariate analysis. The authors retrospectively determined whether 118 (59%) of the EML patients received localized radiotherapy to the site of EML: 42 did and 76 did not. There were no differences in estimated event-free survival between patients who did and did not receive radiotherapy. CONCLUSIONS: Non-skin (group 2) EML appeared to be an independent favorable prognostic factor. Localized radiotherapy to the site of EML at the end of induction chemotherapy did not improve outcome. PMID- 14528098 TI - Targeted radiotherapy with submyeloablative doses of 131I-MIBG is effective for disease palliation in highly refractory neuroblastoma. AB - PURPOSE: Treatment of refractory neuroblastoma remains a significant clinical problem. Targeted radiotherapy with 131I-MIBG has demonstrated antitumor activity in heavily pretreated neuroblastoma patients with recurrent disease. Response rates may be correlated with total radionuclide dose per kilogram body weight delivered, but higher dose levels are associated with protracted grade 4 hematologic toxicity. The optimal method for using single-agent 131I-MIBG for patients with relapsed high-risk neuroblastoma has not been defined. This study was designed to retrospectively determine the clinical response to 131I-MIBG therapy at submyeloablative doses in patients with refractory neuroblastoma and to describe the toxicities. PATIENTS AND METHODS: A retrospective chart review of 20 patients with neuroblastoma treated with 131I-MIBG at the Children's Hospital of Philadelphia from 1988 to 2000 was performed. Demographic data, 131I-MIBG dose delivered, toxicities, and clinical responses were reviewed. RESULTS: A median dose of 9.5 mCi/kg of 131I-MIBG was delivered in 32 courses to 20 patients. Three patients were treated in first complete response, and the remaining 17 patients for residual and/or progressive disease. The objective response rate to the first therapy was 31%, and the remaining patients achieved disease stabilization. In addition, 9 of 11 patients with pain at study entry had significant improvement. Disease response was not correlated with 131I-MIBG dose delivered. No unanticipated toxicities were observed. CONCLUSIONS: Submyeloablative-dose 131I MIBG is an effective and relatively nontoxic method for neuroblastoma disease palliation. Most patients show subjective improvement in pain and/or performance status. Increased availability and experience with 131I-MIBG therapy would benefit a large number of children with end-stage neuroblastoma and no realistic hope for cure. PMID- 14528099 TI - Intensive induction chemotherapy and delayed irradiation in the management of parameningeal rhabdomyosarcoma. AB - PURPOSE: To evaluate local control, event-free survival, and overall survival for patients with parameningeal (PM) rhabdomyosarcoma (RMS) treated with intensive chemotherapy and delayed irradiation. PATIENTS AND METHODS: Thirteen consecutive patients with PM RMS were treated with an institutional protocol from 1992 to 1998 at the University of Washington/Children's Hospital and Regional Medical Center and Deaconess Medical Center. Patients received intensive chemotherapy consisting of vincristine, doxorubicin, cyclophosphamide, ifosfamide, and etoposide prior to radiotherapy. Irradiation was delayed, in contrast to current Intergroup Rhabdomyosarcoma Study Group (IRSG) recommendations. RESULTS: Median follow-up was 39 months. Eleven patients had high-risk features, including five with intracranial extension. All patients responded to the intensive chemotherapy, with 38% exhibiting a complete response and the remaining 62% a partial response. Radiation was administered a median of 21 weeks from initiation of chemotherapy. The Kaplan-Meier estimate of 5-year local control was 92%, with event-free survival and overall survival rates of 83%. CONCLUSIONS: With intensive induction chemotherapy, delayed irradiation for PM RMS does not compromise local control. Event-free survival and overall survival rates compare favorably with recently IRSG trials employing early irradiation. Delaying irradiation allows for intensification of chemotherapy and could permit response based radiation volume and/or dose modifications, which could decrease treatment related morbidity. PMID- 14528100 TI - Analysis of polymorphisms of the vitamin D receptor, estrogen receptor, and collagen Ialpha1 genes and their relationship with height in children with bone cancer. AB - PURPOSE: The authors' objectives were to compare height at diagnosis of children with bone tumors with that of Spanish reference children; to analyze the frequency of the genotypes for the polymorphisms of the vitamin D receptor (VDR), estrogen receptor (ER), and collagen Ialpha1 (COLIalpha1) genes in patients and in healthy controls; and to test the relationship between the genetic markers and height. PATIENTS AND METHODS: Height and weight at diagnosis were measured in 58 osteosarcoma and 36 Ewing sarcoma patients and compared with standards published for Spanish reference children according to sex and age. For the molecular analysis, genetic polymorphisms of the VDR (Fok I, Apa I, and TaqI), ER (Pvu II and XbaI), and COLIalpha1 (Msc I) genes were characterized in 72 osteosarcoma and 53 Ewing sarcomas and in a group of 143 healthy matched children. RESULTS: Osteosarcoma and Ewing sarcoma patients were significantly taller than Spanish reference children. Osteosarcoma patients showed a significantly higher frequency of the Ff genotype for the Fok I polymorphism (VDR gene) than the control group. The odds ratio for this genotype was 1.78, with an increased relative risk of 78% for heterozygous Ff carriers. Among Ewing sarcoma patients, this same genotype was significantly associated with lower height than homozygotes (FF or ff). CONCLUSIONS: Children with bone cancer are significantly taller than the reference population, which may be influenced by the genotype for the Fok I polymorphism of the VDR gene. PMID- 14528101 TI - Clinical trials in pediatric cancer: parental perspectives on informed consent. AB - To better understand parental perceptions of the informed consent process in pediatric oncology clinical trials, 20 parents of newly diagnosed children at two pediatric cancer centers described their perceptions in a semi-structured interview. They recalled well the diagnosis, the general treatment plan, and the statistics of survival and/or cure, but the research nature of the clinical trials, particularly randomization, was not well understood. However, despite the need to assimilate a great deal of information, time pressure to make decisions, and reportedly high levels of distress during the discussions, parents expressed general satisfaction with the informed consent discussions with their pediatric oncology providers. However, half to two thirds of parents felt there had been inadequate discussion of alternatives to the proposed treatment and of the research nature of the protocol. While further study of the informed consent process should be conducted in larger, representative samples, the findings from this pilot study suggest that a goal of future informed consent interventions should be to improve parents' understanding of the research aspects of treatment. It is critical to parents' ability to provide informed consent that they feel satisfied that they know alternatives to proposed treatment and that they understand the randomization of treatments, which is the gold standard of clinical trials in pediatric oncology. PMID- 14528102 TI - Deletional mutation of the external domain of the human granulocyte colony stimulating factor receptor in a patient with severe chronic neutropenia refractory to granulocyte colony-stimulating factor. AB - Severe chronic neutropenia (SCN) is characterized by a profound neutropenia, which mostly presents during the neonatal period. The precise genetic basis of SCN remains elusive. Acquired somatic mutations involving the carboxy-terminus of the G-CSF receptor (G-CSFR) have been found, often in association with myelodysplastic syndrome. The authors describe a girl with SCN who did not respond to pharmacologic doses of filgrastim. Genetic analysis of bone marrow and germline cells revealed a 182-bp deletion in the extracellular domain of the G CSFR. Co-precipitation studies showed an association between the wild-type and mutant G-CSFR, confirmed by their co-localization by confocal microscopy. Coexpression of the mutant receptor inhibited the wild-type response in Ba/F3 cells. These findings establish a novel constitutional defect in the G-CSFR that supports a partial dominant negative mechanism for receptor dysfunction in SCN. PMID- 14528103 TI - Low-dose versus high-dose immunoglobulin for primary treatment of acute immune thrombocytopenic purpura in children: results of a prospective, randomized single center trial. AB - PURPOSE: To investigate the efficacy and side effects of two different intravenous immunoglobulin (IVIG) dose regimens for the initial treatment of childhood acute immune thrombocytopenic purpura (ITP). METHODS: Thirty-four consecutive patients with a clinical diagnosis of acute ITP and a platelet count below 20x10(9)/L were randomized to receive either 1 g/kg body weight (n=17; group A) or 0.3 g/kg body weight (n=17; group B) IVIG per day for 2 consecutive days (total dose 2 g/kg and 0.6 g/kg). RESULTS: Fifteen of the 17 patients (88.2%) in group A and 13 of the 17 patients (76.5%) in group B achieved a platelet count of more than 20x10(9)/L within 72 hours. The increase in platelet counts on day 2 and 3 was more pronounced in the high-dose group. Two patients in the high-dose group and four in the low-dose group were non-responders. Chronic disease occurred in three patients receiving 2 g/kg IVIG and in five patients receiving 0.6 g/kg IVIG. Side effects of IVIG administration were more common in the high-dose group. CONCLUSIONS: The present study showed that platelet counts increased more rapidly after high-dose IVIG administration within the first 72 hours, although a platelet count of more than 20x10(9)/L can be achieved also with low-dose IVIG in most children with acute ITP. For patients with very low platelet counts, doses higher than 0.6 g/kg seem, therefore, to be more effective. PMID- 14528104 TI - Continuous veno-venous hemofiltration may improve survival from acute respiratory distress syndrome after bone marrow transplantation or chemotherapy. AB - PURPOSE: Acute respiratory distress syndrome (ARDS) may result from immunologic activity triggered by irradiation and/or chemotherapy. Hemofiltration removes plasma water and soluble components below 25 kilodaltons. The authors hypothesized that early hemofiltration might attenuate the inflammatory component of ARDS, resulting in increased survival in immunocompromised children and young adults. METHODS: Ten children (6 bone marrow transplantation, 3 chemotherapy, 1 lymphoma/hemophagocytosis) with ARDS (Pao2/Fio2 94 +/- 37 torr) received early continuous veno-venous hemodiafiltration as adjunctive therapy for respiratory failure, regardless of renal function. Six children had normal urine output and initial serum creatinine (range 0.1-1.2 mg/dL); four had renal insufficiency (initial creatinine 1.7-2.4 mg/dL). Hemofiltration was instituted coincident with intubation. Respiratory failure was precipitated by Enterobacter sepsis in two patients and by Aspergillus in one. RESULTS: Hemodiafiltration was performed for 13 +/- 9 days. A high rate of clearance was achieved (52 +/- 17 mL/min/1.73 m2). Duration of mechanical ventilation was 14 +/- 9 days. Nine of the 10 children were successfully extubated; 8 survived. CONCLUSIONS: Early hemofiltration may improve survival from ARDS following bone marrow transplantation or chemotherapy. Possible mechanisms include strict fluid balance, immunomodulation through filtration of inflammatory constituents, and immunomodulation through intensive extracellular water exchange that delivers biochemicals to organs of metabolism as well as the hemofilter. PMID- 14528105 TI - Skip metastasis in osteosarcoma. AB - Skip metastasis is rare in high-grade osteosarcoma and indicates a very poor prognosis. The authors describe two patients who had high-grade osteosarcoma and skip metastasis. MRI detected the skip lesion in both patients, and both lesions were confirmed by histology. The patients were treated with adjuvant chemotherapy and radical surgical resection. Both are alive and free of disease 3 and 6 years from diagnosis. PMID- 14528106 TI - Favorable response to treatment of a child with T-cell-rich large B-cell lymphoma presenting with liver failure. AB - The authors describe the successful management of a child with T-cell-rich large B-cell lymphoma (TCRBCL) involving the lymph nodes and liver, causing severe hepatic dysfunction. After immunohistochemical confirmation of the diagnosis, the patient was treated initially with low-dose, non-hepatotoxic chemotherapy and irradiation to the porta hepatis. Chemotherapy was gradually escalated to intensified B cell-lymphoma treatment regimens (CHOP, CYVE, COPAdM) as liver function improved. Despite prompt and notable response in the lymph nodes, the intrahepatic nodular lesions persisted radiologically. Liver function tests normalized during the course of treatment. However, radiologic studies of the liver showed abnormal regeneration with a predominant left lobe overshadowing a tiny right lobe. The patient remains in remission more than a year after completion of treatment. Judicious use of chemotherapeutic agents with or without local radiotherapy may influence the outcome and survival in children with TCRBCL involving the liver and causing abnormal hepatic function. PMID- 14528107 TI - Anemia and hepatosplenomegaly as presenting features in a child with rickets and secondary myelofibrosis. AB - Anemia and hepatosplenomegaly are common reasons for referring a child to a pediatric hematologist or oncologist. Among the many causes for these findings is severe rickets, which has been shown to be associated with secondary myelofibrosis and myeloid metaplasia. The authors present the case of an infant with severe rickets and secondary myelofibrosis and review the differential diagnosis of hepatosplenomegaly from the viewpoint of the pediatric hematologist/oncologist. PMID- 14528108 TI - Radiation therapy in the treatment of Gorham disease. AB - An adolescent male with Noonan syndrome presented with a nonhealing fracture of the proximal right humerus. Over a 7-month period there was progressive loss of bone in this region, resulting in a flail arm at 9 months. Radiographic review was consistent with Gorham disease. In addition, there was significant bleeding in the soft tissues and pain. Radiation was delivered. Seven years passed, until the patient re-presented with right hemithorax near collapse secondary to chylothorax. A chest tube was placed with temporary relief, but significant effusion remained. Radiation was again administered, and by the end of therapy the chest tube was removed. The effusion has not recollected at last follow-up, which is now 6 months. The use of radiation in the treatment of Gorham disease has been demonstrated to have excellent palliative ability. PMID- 14528110 TI - Significance of bone marrow examination in the diagnostic process of paraspinal mass in children: a case report. AB - Spinal cord compression is a rare but serious complication of non-Hodgkin lymphoma or leukemia. Biopsy of the mass with or without laminectomy would be necessary for diagnosis in patients presenting with isolated paraspinal mass in the absence of other overt clinical symptoms or findings or laboratory abnormalities that would specifically suggest leukemia or lymphoma. The authors describe a 6-year-old girl with symptoms of spinal cord compression due to a paraspinal mass who had undergone surgical intervention; she was later shown to have acute B-cell lymphoblastic leukemia. A bone marrow aspiration should be considered in such patients to avoid unnecessary surgical intervention. PMID- 14528109 TI - Misinterpretation of a Calvert-derived formula leading to carboplatin overdose in two children. AB - Carboplatin is currently recommended to be dosed according to renal function. In adults, dosing by the Calvert formula is based on evidence that carboplatin clearance closely parallels glomerular filtration rate. Several studies have attempted to validate the Calvert formula and its derivations in pediatrics, but no final consensus has been achieved. As a result, different versions of the original formula exist in the pediatric literature. Other factors may also contribute to confusion when applying the formula to young patients, including the manner in which renal function is measured and reported. We describe how misinterpretation of the Calvert formula resulted in carboplatin overdosing in 2 pediatric patients with high-risk neuroblastoma undergoing peripheral blood stem cell transplantation. Measures to avoid such errors have been instituted. PMID- 14528111 TI - Fludarabine and once-daily intravenous busulfan for allogeneic bone marrow transplantation for Chediak-Higashi syndrome. AB - An HLA-identical sibling bone marrow transplant was done for a patient with Chediak-Higashi syndrome. The preparative regimen included intravenous fludarabine (40 mg/m2/dx4) and busulfan (130 mg/m2/dx4). Busulfan was given once daily. Pharmacokinetic studies showed the area under the concentration-time curve of the once-daily intravenous busulfan was similar to that seen with the total daily dose administered with an every-6-hourly regimen. Toxicity was minimal. Myeloid engraftment occurred on day +17 and donor chimerism was complete. Fludarabine and once-daily intravenous busulfan is well tolerated and is adequate for engraftment of sibling transplant in Chediak-Higashi syndrome. PMID- 14528112 TI - Fatal fulminant hypereosinophilic syndrome with a petechial rash in a 16-year-old girl. AB - Peripheral blood eosinophilia and eosinophilic tissue infiltration characterize the hypereosinophilic syndrome, which if untreated can be fatal. Its manifestations are protean. Often a diagnosis of exclusion, it presents diagnostic challenges. A fatal case of hypereosinophilic syndrome in a 16-year old girl with petechiae, edema, urticaria, and diffuse erythema unfolded over 2 weeks. The histopathology of the petechiae demonstrated an eosinophilic and mixed cell interstitial and perivascular infiltrate and microthrombi in vessels. There was no evidence of T-cell clonality. Despite treatment with cyclosporine and methylprednisolone, she died. The age group; fulminant, fatal course without neoplasia; striking eosinophilia; and petechial rash are notable. PMID- 14528113 TI - Simplified method to assay total plasma peroxidase activity and ferriheme products in sickle cell anemia, with initial results in assessing clinical severity in a trial with citrulline therapy. AB - A method using dianisidine is described to measure promptly the total plasma peroxidase (POX) activity of methemoglobin and other ferrihemes. Methemoglobin (35 mg/dL) is used as POX standard. Three-minute POX activities and total POX concentrations measured by a classic benzidine method were compared in a three patient trial with citrulline supplementation. High POX values became progressively lower. In two of the patients, 3-minute POX activities were reduced more than total concentrations. Oral citrulline reduced harmful plasma ferriheme levels. Free ferriheme also exhibited POX activity. POX levels may be useful to judge severity in sickle cell anemia and to monitor therapeutic efficacy. PMID- 14528114 TI - Cold agglutinin disease after hepatitis A infection in a child. PMID- 14528115 TI - The numbers of our lives. PMID- 14528116 TI - The management of severe malaria. PMID- 14528117 TI - Central venous catheters in the pediatric intensive care unit: access versus infection. PMID- 14528118 TI - Ventilator-associated pneumonia in intubated and mechanically ventilated children: challenges in diagnosis. PMID- 14528124 TI - Breathing lessons: basics of oxygen therapy. PMID- 14528123 TI - Understanding gastroesophageal reflux disease. PMID- 14528125 TI - Mandatory nurse/patient ratios: a good idea or not? PMID- 14528126 TI - Patient education series. Low back pain. PMID- 14528127 TI - Salary survey. PMID- 14528128 TI - Assessing fetal heart sounds. PMID- 14528129 TI - Low-country lessons. PMID- 14528130 TI - Infusing without infecting. PMID- 14528131 TI - Acute coronary syndrome. PMID- 14528132 TI - The political and policy implications of a rating system of community capacity to improve population health. AB - This article discusses the political and policy implications of a potential rating system of community capacity for health improvement. Evidence suggests that such a rating system will have little effect in the current health care environment unless it is directly related to the health care dollar. Community change agents and stakeholders, including the growing population of older adults, will perceive a rating system to have value only if the defined purpose meets their interests and needs, and if the rating system results prove useful for decisions that affect the bottom line in working on issues like health improvement and capacity. PMID- 14528133 TI - Developing community indicators to promote the health and well-being of older people. AB - As the US population ages and the number of older people who are "aging in place" increases, communities will face new opportunities and challenges in responding to this population's desires and needs. Qualitative research was conducted to inform the development of a model of an "elder-friendly community" and a set of indicators to measure and help improve community capacity to promote the health and well-being of older residents. Focus groups were conducted in four US cities with younger and older adults and community leaders who identified attributes that make a community a good place in which to grow old. The group interactions were videotaped, transcribed, and analyzed to identify common themes. Results were remarkably similar across sites. Participants said that a community could be considered elder-friendly if it helped older residents continue active participation in the community, sustain their independence, and reduce the risk of isolation. A model of an elder-friendly community, along with corresponding indicators, was created on the basis of focus group results. These tools are being tested in 10 pilot communities to determine their usefulness for measuring older people's health and well-being, prioritizing aging issues, and stimulating and informing action strategies to improve community "elder-friendliness." PMID- 14528134 TI - Comparison of Health Buddy with traditional approaches to heart failure management. AB - The purpose of this pilot study was to (a) determine the feasibility of providing a heart failure disease management program through an in-home telehealth communication device (Health Buddy) and (b) compare the effectiveness of the Health Buddy with traditional home management strategies (telephonic, home visit) in achieving selected patient outcomes (self-efficacy, functional status, depression, and health-related quality of life). Ninety participants completed the study through 2 months. Thirty percent of participants were either eliminated prior to or withdrawn after enrollment from the study based on Health Buddy issues. A mixed model ANOVA revealed those who received telephonic disease management experienced decreased confidence in their ability to manage their heart failure whereas all other groups experienced increased confidence. Further ANOVA analyses indicated improvement over time with no group differences for functional status, depression, or health-related quality of life. These findings suggest that delivering a disease management program through a telehealth communication device is feasible and may be as effective as traditional methods. PMID- 14528135 TI - HIV/AIDS case profile of African Americans: guidelines for ethnic-specific health promotion, education, and risk reduction activities for African Americans. AB - There are discrepancies in health care services for the poor and ethnic minorities in the United States. Within the past decade widespread concerns regarding the need to reform the nation's health care services, including the problem of human immunodeficiency virus/acquired immune syndrome (HIV/AIDS) among African Americans has continued. These inequalities have been the cornerstone of the U.S. Healthy People 2010 national priority objectives. The objectives focus on health and social outcomes such as low quality of life and mortality rates, poverty, lack of accessibility to and appropriateness of care, and the prevalence of certain degenerative conditions and infectious diseases. The dearth of preventive health services for the high-risk groups, particularly children, adolescents, young adults, and older African American adults undermines early intervention efforts, including prompt HIV/AIDS identification and diagnosis, prevention education, health promotion, effective substance abuse treatment, and counseling services. This work reviews the magnitude of HIV/AIDS among African Americans between 1996 and 1999 by race/ethnicity, gender, and age groups. It also addresses the major factors responsible for the continued upward trend in the distribution and rate of infectiousness of HIV/AIDS among African Americans. The study recommends and discusses culturally sensitive and ethnic-specific intervention strategies for the prevention and control of HIV/AIDS among African Americans. PMID- 14528136 TI - Outreach to multiethnic, multicultural, and multilingual women for breast cancer and cervical cancer education and screening: a model using professional and volunteer staffing. AB - Reaching low-income women from diverse ethnic and cultural communities for the purpose of breast and cervical cancer education and screening is a challenge. This article describes an approach used by Center for Healthy Aging in Santa Monica, California, to implement three projects aimed at Hispanic and African American women to encourage early intervention behaviors. Hispanic and African American volunteers and professional staff work together to establish trusting relationships, join forces with community leaders and develop collaborative approaches to achieve the goal of screening about 1,000 underserved women annually and providing education for an additional 5,000 women. Incorporated in the article are lessons learned about working with this population, the communities in which they live and the use of volunteers for these and other projects. PMID- 14528137 TI - Early intervention screening for family caregivers of older relatives in primary care practices: establishing a community health service alliance in rural America. AB - The Maine Primary Partners in Caregiving project provides a prime example of how disparate community health, social service, and higher education institutions can build a successful rural service alliance for the purposes of screening for family members experiencing stress during the provision of care to impaired older relatives. Community primary care practices are featured as prime sites for the early identification of elder caregivers experiencing stress and burden. Initial project results and implementation challenges as well as recommended strategies for nurturing such community partnerships are presented. PMID- 14528138 TI - Collaborative community-based research and innovation. AB - This work describes the results and lessons learned from a community-academic partnership to research the effect of training on the capacity of rural home care aides to care for older persons with dementia. The research study increased the aides' knowledge of Alzheimer's and related disorders and, for one group, the aides' satisfaction. The authors used content analysis to identify barriers and facilitators of success of the research partnership. These factors are discussed and placed within a framework of innovation concepts. PMID- 14528139 TI - An international approach to community health care for older adults. AB - Worldwide population ageing, concomitant increases in disability rates, and changes in family health care systems require an examination of current service delivery to optimize use of societal resources in the future. This article examines a community health care approach suggested by research conducted by the World Health Organization Kobe Centre for Health Development (WKC). The WKC approach, which uses a cross-national perspective, envisions a community health care system that integrates health and social services and spans health promotion, primary care, and long-term care. Prototypical approaches for organizing community health care include communal, marketplace, case management, and managed care. The ramifications of these approaches are examined from the perspectives of the older adult, the family, and formal service providers. PMID- 14528147 TI - [Normal spirometric values]. PMID- 14528148 TI - [Medical demography: the respiratory physicians among others]. PMID- 14528149 TI - [Lung volume reduction surgery for emphysema: still many unsolved physiological issues]. PMID- 14528150 TI - [Severe asthma: can we do better?]. PMID- 14528151 TI - [New tables for the compensation of employment-related respiratory diseases]. PMID- 14528152 TI - ["Inhalotherapy" in the province of Quebec]. PMID- 14528153 TI - [High dose rate brachytherapy: a potentially curative treatment for small invasive T1N0 endobronchial carcinoma and carcinoma in situ]. AB - INTRODUCTION: High dose rate brachytherapy (HDR-BT) is an option for treatment of small invasive endobronchial carcinoma and carcinoma in situ. This retrospective study describes the results of 33 consecutive patients treated with curative intent and followed up for more than one year. METHODS: Between July 1994 and October 1999, 35 tumours were treated with HDR-BT alone using a standard protocol delivering 6 fractions of 5 Gy delivered across 1cm from the catheters over 3 to 6 weeks. In 31 patients, surgical treatment was ruled out because of histology (in situ carcinoma), history of pneumonectomy or significant co-morbidity. Two patients were treated for positive resection margins following pneumonectomy. All the tumours were Tis or T1 N0. RESULTS: The locations of tumours were: trachea - 2, main bronchus - 5, lobar - 20, and segmental - 8. Only one catheter was needed in 15 cases, 2 in 13 cases, 3 in 6 cases and 4 in 1 case. The median follow-up was 17 months (range, 5-53 months). The recurrence-free rate was 94.3% at 2 months and 86.2% at 6 months after the treatment. 15 patients (45.5%) developed local recurrence at a median time of 9 months. Three patients developed metastases. The 1-year and 2-year overall survival rate were 71.4% and 53.8% respectively and specific survival rates were 69.4% and 59%. One patient developed an acute pneumothorax, and late complications included 6 infections and 12 bronchial stenoses. There were no episodes of haemoptysis nor lethal complications. CONCLUSIONS: With strict selection criteria, HDR-BT can be a curative treatment for early invasive or in situ endobronchial carcinoma without serious toxicity. It is therefore a good alternative treatment with curative intent for inoperable patients. PMID- 14528154 TI - [Vital capacity and peak expiratory flow rates in a North-African population aged 60 years and over: influence of anthropometric data and parity]. AB - INTRODUCTION: The aims of this study were to calculate predictive equations for respiratory variables (FVC, FEV1, FEF25%-75%, and PEFR) in elderly north-African people and to study the effect of parity on the respiratory function. METHODS: FVC and forced expiratory flows were measured in a "healthy" sample of 186 subjects aged 60 years and over (66% women). The measurements were made in health centres using portable spirometers. Predictive equations were determined by the technique of multiple linear regression using standing height and age as independent variables. The effect of parity on ventilatory function was studied by comparing two groups defined by a parity greater or less than 4. RESULTS: The respiratory variables measured in this elderly north-african population were significantly different from those established references ranges. CONCLUSIONS: High parity is associated with a significant reduction in peak flow rate. PMID- 14528155 TI - [Demography of lung specialists in France]. AB - INTRODUCTION: This article analyses the medical demography data in France, particularly the situation concerning lung specialists. METHODS: Study of the published data, particularly institutional reports (Ministry of Health, Sickness Insurance). RESULTS: The demography is characterised by important differences between specialties, regions, and salaried and private practice, as well as the effects of ageing and the increasing numbers of female practitioners. Lung specialists are, on average, younger and more evenly distributed between the different types of practice. Nevertheless they risk experiencing the same difficulties as other specialties in responding to the needs of the French population. The principle proposals of the public authorities concern the restriction of numbers, the regulation of established specialists and new methods of access to the speciality. CONCLUSIONS: Medical demography has a major role in the future of the French health care system. PMID- 14528156 TI - [Physiological effects of lung reduction surgery: the contribution of an animal model of emphysema]. AB - INTRODUCTION: Lung volume reduction surgery (LVRS) is a technique largely used for palliation of symptoms in selected patients with emphysema. Despite some encouraging early results, it is often still regarded as an experimental technique due to the significant associated mortality and unpredictable results. STATE OF ART: The aim of this study was to explore the pathophysiological basis of benefit from LVRS when applied to a hamster model of emphysema induced by elastase and thus allow better selection criteria of patients being considered for this treatment. PERSPECTIVES: Despite a positive effect on pulmonary elastic recoil pressures, LVRS did not, unlike its effects in humans, improve airway obstruction in the emphysematous hamsters. Differences in chest wall mechanics may explain these contrasting findings. Adaptation of the intrinsic and cellular properties of the diaphragm that are seen in emphysema are preserved after LVRS. CONCLUSIONS: Despite differences with emphysema in humans, this hamster model supports the importance of the residual volume/total lung capacity ratio when selecting patients with emphysema for LVRS. The positive outcome of this intervention on the dynamics of the diaphragm is not compromised by modifications undergone by the diaphragmic myocytes. PMID- 14528157 TI - [Psychosocial support for pulmonary rehabilitation in patients with Chronic Obstructive Pulmonary Disease]. AB - INTRODUCTION: This paper focuses on recent studies examining psychosocial support provided in pulmonary rehabilitation programmes for patients with Chronic Obstructive Pulmonary Disease (COPD). STATE OF ART: The literature shows the impact that psychological variables have on COPD consequences and on compliance with treatment. These parameters influenced rehabilitation outcomes. Psychosocial support, such as group discussions, reduces incapacity (dyspnoea control) and handicap situations (reductions in anxiety and depression, improvements in coping strategies, autonomy and social participation). PERSPECTIVES: Further studies are needed to optimise psychosocial input particularly by combining it with education sessions. CONCLUSIONS: Psychosocial support is nowadays recommended as part of pulmonary rehabilitation programmes for patients with stable COPD. It complements the other components of exercise, breathing techniques and health education. PMID- 14528158 TI - [Sleep apnea syndrome in the elderly]. AB - INTRODUCTION: Sleep apnoea/hypopnoea syndrome (SAHS) is distinct in the elderly as age-related co-morbidity may be aggravated by sleep-related breathing disorders or may mask typical clinical symptoms. STATE OF ART: The SAHS prevalence in the elderly varies from one study to another, with an average rate of 25% that might increase according to the importance of associated pathologies. Clinical symptoms tend to be the neurological manifestations of excessive daytime sleepiness and cognitive deterioration. PERSPECTIVES: Treatment of SAHS should include weight reduction strategies if obesity is present. There is no role for surgery in this age group. On the other hand, adjustable oral appliances can be used depending on the severity of the SAHS, but nasal continuous positive airway pressure (CPAP) remains the most effective therapy available. Acceptance of and compliance with CPAP treatment is as good in this age group as in younger patients, with remarkable effectiveness in terms of improvement in daytime sleepiness and cognitive function. However, the effectiveness of treatment for SAHS is related to the level of symptoms present and this should be taken into account when making a therapeutic decision. CONCLUSIONS: These results fully justify treatment of SAHS in the elderly, but the decision to treat should be governed by the level of clinical symptoms from SAHS. PMID- 14528159 TI - [Immune responses in broncho-pulmonary cancer]. AB - INTRODUCTION: The anti-tumoral immune response is essentially cellular and numerous studies have been conducted with the aim of augmenting it from the therapeutic aspect. STATE OF ART: Activation of the immune system has been reported in the early stages of bronchial carcinoma. On the other hand numerous studies have demonstrated a degree of immuno-suppression in patients with advanced cancer prior to any immuno-suppressant treatment, particularly an alteration of cell mediated immunity. The presence of white cells, infiltrating the tumour is an indication of an active host response to the tumour. Several para-neoplastic syndromes associated with the presence of auto-antibodies have been described, particularly in small cell bronchial carcinoma. Other antibodies not associated with para-neoplastic syndromes have also been found, principally directed against the tumour suppressant gene P53. Non-specific active immuno therapy (BCG, interferons, interleukins.), passive immuno-therapy (monoclonal antibodies) and adoptive immuno-therapy have been studied for many years without encouraging results. Specific active immuno-therapy (anti-tumour vaccines and gene therapy) is the route being most actively explored at present. PERSPECTIVES: Correct targeting of patients likely to benefit from immuno-therapy, specificity and effectiveness of the agents, the expression of transfected genes and the risk of infection and dissemination during clinical trials are all problems currently under investigation. CONCLUSIONS: Manipulations of the immune response have been and are being undertaken with the aim of improving its effectiveness. Anti-tumour vaccines and gene therapy are the routes most studied. PMID- 14528160 TI - [Health-related quality of life measurement: a critical reading guide]. AB - INTRODUCTION: Quality of life is rarely assessed in the pulmonary function laboratory. Quality of life, or at least some of its domains, should be directly measured. STATE OF ART: Quality-of-life questionnaires developed according to established protocols can be very useful in clinical practice. In clinical studies, the choice of a questionnaire depends on its function (evaluative or discriminative) which should be supported by its measurement properties (validity, reliability, responsiveness). PERSPECTIVES AND CONCLUSIONS: The methodology of critical appraisal of an article about quality of life proposed in this reading guide are in keeping with the principles of evidence-based medicine. PMID- 14528161 TI - [Early nasal ventilation in amyotrophic lateral sclerosis: impact on survival and quality of life (the VNP-SLA study)]. PMID- 14528162 TI - [What is a confidence interval?]. PMID- 14528163 TI - [What does "power of the study" mean? How to compute the power of a study? How to compute the number of subjects needed for a trial?]. PMID- 14528164 TI - [Intra-cavity aspergilloma complicating necrotising sarcoid granulomatosis]. AB - INTRODUCTION: Necrotising sarcoid granulomatosis is a rare clinical entity, characterised by the development of nodular pulmonary opacities, the diagnosis if which is based on the identification of characteristic pathological lesions. CASE REPORT: A 44-years-old male smoker presented with cough and fever. With the exception of a few basal crepitations clinical examination was normal. The chest x-ray showed multiple areas of alveolar consolidation scattered throughout both lung fields, some of which were cavitated. The thoracic CT scan confirmed the presence of cavitating nodules and hilar and mediastinal lymphadenopathy. Samples for mycobacteria were negative. A search for anti-neutrophil cytoplasmic antibodies was negative. Fibreoptic bronchoscopy was normal but transbronchial biopsies revealed giant cells. Lung biopsies taken by video thoracoscopy showed non-caseating granulomata with areas of fibrinoid necrosis and perivascular infiltration with lymphocytes, histology characteristic of necrotising sarcoid granulomatosis. Resolution occurred without treatment. The parenchymal lesions regressed leaving residual cavities. Four years later, following an episode of haemoptysis, the patient was found to have a squamous cell carcinoma (T1N0M0) and aspergillomas in the pulmonary cavities. CONCLUSIONS: Our observation demonstrates that the cavities of necrotising sarcoid granulomatosis may be complicated by aspergillomas. PMID- 14528165 TI - ["Post traumatic" leptospirosis complicated by acute respiratory insufficiency]. AB - INTRODUCTION: Leptospirosis is a rare cause of alveolar haemorrhage. The diagnosis is often delayed particularly when the mode of infection is atypical. These serious complications require prompt antibiotic treatment. CASE REPORT: A 21-year-old man was involved in a road accident and found lying unconscious in a roadside ditch containing stagnant water. Ten days later he presented with bilateral interstitial pneumonia and rapidly increasing hypoxaemia associated with cholestasis and liver cell necrosis. Broncho-alveolar lavage revealed alveolar haemorrhage. There was satisfactory resolution following antibiotic therapy. CONCLUSIONS: The diagnosis of leptospirosis was considered initially despite negative serology (Martin and Petit) and confirmed by sero-conversion 20 days after the onset of symptoms. PMID- 14528166 TI - [Costo-vertebral hydatid disease: the role of MRI]. AB - INTRODUCTION: Hydatid disease of bone is very rare accounting for 0.5% - 3% of all localisations. CASE REPORT: We report the case of a woman of 30 years who presented with posterior chest pain. The chest radiograph showed an extra pulmonary mass associated with a lytic rib lesion. Thoracic ultrasound revealed a multiloculated soft tissue and fluid mass. CT scan showed a fluid filled costo vertebral cyst. MRI identified extension into the spinal canal. Pathological examination of the surgically resected material confirmed the diagnosis. CONCLUSIONS: This observation emphasises the role of MRI in assessing the extent of hydatid cysts of the posterior chest wall and in particular invasion of the spinal canal. PMID- 14528168 TI - [Constrictive pericarditis due to asbestos exposure]. AB - INTRODUCTION: Constrictive pericarditis is a rare complication of asbestos exposure and few cases have been reported in the literature. CASE REPORT: We report two cases of constrictive pericarditis in subjects previously exposed to asbestos. The first case, a 62 years old man, had occupational asbestos exposure whilst working for seven years in an electric plant 23 years before the diagnosis. The second case, a 76 years old man, had worked 21 years as a lagger up until 20 years before. The initial presentation in both cases was of sub-acute right heart failure. Both underwent pericardectomy which revealed pericardial thickening due to collagen fibrosis. Both patients died, one and five years respectively after surgery. Eight other cases of pericardial effusion and/or thickening, some with calcification, have been reported in association with previous asbestos exposure. Most of these cases had coexisting pleural lesions. CONCLUSIONS: As the prognosis is guarded (three of the eight reported cases died), making an early diagnosis is desirable. PMID- 14528167 TI - [Amyloidosis and pleural calcification]. AB - INTRODUCTION: Pulmonary diseases during the course of generalised amyloidosis are principally represented by tracheobronchial involvement and diffuse parenchymal localizations. CASE REPORT: The authors report the case of a 66-year-old woman presenting with pleural amyloidosis in the context of generalised amyloidosis. Thoracoscopy performed in the investigation of recurrent transudative pleural effusions found evidence of an inflamed parietal pleura with areas of calcification. Pleural biopsies confirmed amyloid infiltration pleural ossification. Talc pleurodesis was performed. CONCLUSIONS: The authors' conclusion is that, meeting an unexplained pleural effusion even transudative during the course of a generalised amyloidosis, the thoracoscopy is the diagnosis key test as it allows moreover a pleural pleurodesis to be performed. PMID- 14528169 TI - [Familial diffuse interstitial fibrosis of prolonged onset worsening during pregnancy]. AB - INTRODUCTION: Familial idiopathic interstitial pulmonary fibrosis is rare. In this case report the diagnosis was confirmed histologically in four members of the same family. CASE REPORT: A woman whose father and two paternal uncles had developed pulmonary fibrosis was hospitalised from birth on account of delayed growth and dyspnoea. At the age of one year an increase in dyspnoea and the development of hypoxaemia and diffuse interstitial shadowing led to a surgical lung biopsy. The histological diagnosis was idiopathic interstitial fibrosis. Immunosuppressive treatment for one year led to clinical improvement with relief of the hypoxaemia but persistence of the interstitial shadowing. A pneumothorax at the age of 15 required pleurectomy. The clinical state remained stable with a restrictive ventilatory defect up to the age of 26 when respiratory insufficiency developed in the course of pregnancy. The outcome following delivery was severe respiratory failure complicated by pulmonary arterial hypertension leading to death the following year. CONCLUSIONS: This case is distinguished by a histological diagnosis in four members of a family of whom one was an infant, the prolonged stabilisation after immunosuppressive therapy and the possible role of pregnancy in the progression. PMID- 14528170 TI - [The discovery of spirometry: back across the Channel]. PMID- 14528171 TI - [Bilateral pneumothorax: a unique observation]. PMID- 14528172 TI - [Pulmonary function tests for the assessment and monitoring of asthma in children above 3 years of age]. PMID- 14528174 TI - Overcoming obstacles to weight loss. What to do when the scale won't budge. PMID- 14528175 TI - Testing for human papillomavirus during a Pap test. PMID- 14528176 TI - Potent antioxidant improves diabetic nerve damage. PMID- 14528177 TI - Quick benefits seen for women who quit smoking. PMID- 14528178 TI - Low-carbohydrate diets. What are the pros and cons? PMID- 14528181 TI - Intense health anxiety. Preoccupation with illness. PMID- 14528179 TI - Endocarditis. When infection attacks your heart. PMID- 14528182 TI - I've heard there's a nail polish for nail fungus. Does it work? PMID- 14528183 TI - The flu vaccine has been available for four decades and yet the death rate from the flu remains high. Why? PMID- 14528184 TI - How often should I have a skin examination? PMID- 14528185 TI - Mayo Clinic office visit: issues in adult immunization. PMID- 14528186 TI - Patient experiences with atrial fibrillation and treatment with implantable atrial defibrillation therapy. AB - BACKGROUND: Patient perspectives about their illness experiences, symptoms, and treatment are essential aspects of quality of life and provide direction for patient and provider decision making regarding innovative therapies such as implantable devices for arrhythmia. PURPOSE: The purpose of this qualitative study was to describe: 1) the experience of patients living with symptomatic, drug-refractory atrial fibrillation (AF) and 2) patient experiences and acceptance of treatment with the implantable cardioverter defibrillator (ICD) with atrial therapies (ICD-AT) including ventricular and atrial defibrillation therapy. PARTICIPANTS: Subjects were 3 women and 8 men, 35 to 80 years of age, who received the Medtronic Jewel AF 7250 ICD-AT as therapy for recurrent, drug refractory AF, had a history of AF for 3 to 20 years and had experienced multiple treatment modalities including frequent external cardioversion in an effort to control their AF. METHODS: A semi-structured interview addressed experiences of symptoms and prior treatment for AF and experiences, concerns, and perceived benefits of the ICD-AT. Interviews were recorded and transcribed verbatim. Narratives were coded and categorized using Atlas Ti(R) software. Qualitative interpretive analysis methods were used to identify key themes. RESULTS: Before ICD-AT, patient themes focused on AF that was: 1) misdiagnosed, minimized, and poorly treated; 2) distressful because of frequent and intense AF symptoms (fatigue, dizziness, shortness of breath, and anxiety) before ICD-AT; 3) limiting of activities of daily living; 4) associated with distress from enduring previous treatment; and 5) associated with the continuous pursuit of successful treatment and maintenance of normalcy. Decision making regarding ICD-AT therapy included weighing symptom or treatment distress versus anticipated risks or benefits, hope for better outcomes, and lack of options. After ICD-AT, themes included positive perceptions of the device because of AF symptom relief, ability to resume normalcy, and medication tolerance; incorporation of shock experiences into life routines; and patient suggestions regarding preparation and social support. IMPLICATIONS: Symptoms of AF have a major negative impact on overall quality of life. Treatment with the ICD-AT confers a sense of security and reduced symptom distress. Greater provider attention to patient preparation and facilitating social support are important for future ICD-AT patient care. PMID- 14528187 TI - Posttraumatic stress disorder in first-time myocardial infarction patients. AB - OBJECTIVES: The objectives of this study were to investigate the prevalence of posttraumatic stress disorder in patients with a first myocardial infarction compared with a random sample of healthy controls and to determine variables associated with the disorder. DESIGN: A questionnaire was distributed to 112 consecutive patients 4 to 6 weeks after infarction and to 115 healthy controls selected randomly from the general population. Objective clinical measures were obtained from the patients' medical records. RESULTS: Twenty-five (22%) patients qualified for a diagnosis of posttraumatic stress disorder (PTSD) compared with 8 (7%) controls with patients being more than a three-fold (OR: 3.84; 95% CI: 1.65 to 8.94) risk of having the disorder. When adjusting for other variables, the risk was reduced to above a two-fold risk (OR: 2.71; 95% CI: 0.99-7.41). In patients and controls, depression and neuroticism were associated with a diagnosis of PTSD adjusting for other variables. In patients, anxiety was associated with a diagnosis of PTSD adjusting for other variables. Left ventricular ejection fraction and symptoms of angina pectoris were not related to a diagnosis of PTSD in the patient group. CONCLUSIONS: Given that previous research has shown that persons with PTSD are at increased risk of cardiovascular diseases, cardiac patients with the disorder may be at a higher risk of recurrent cardiac events. Although longitudinal studies are needed to confirm such a relationship, this disorder should not be overlooked because of its potential role in reinfarctions and mortality. PMID- 14528188 TI - The coronary artery bypass experience: gender differences. AB - OBJECTIVE: The primary purpose of this study was to examine differences between women and men on physical, social and psychological domains of health quality of life before, 1 month after, and 3 months after coronary artery bypass surgery. DESIGN: A prospective, longitudinal design was used. SETTING: The study was conducted at a Midwestern, 500-bed community hospital with an ongoing cardiothoracic surgical program. PARTICIPANTS: Forty pairs of women and men matched on age within 5 years and body surface area within 0.1m 2. INSTRUMENTS: The physical, social and psychological domains of health quality of life were assessed using the following instruments: Ferrans and Powers Quality of Life Index, Specific Activity Scale, Symptom Scale, Profile of Mood States, Overall Health Rating Index, and Personal Resource Questionnaire. RESULTS: Both women and men improved on physical and psychological measures following coronary artery bypass surgery. Compared with men, women reported more shortness of breath and depression and lower ratings of activity, vigor, and overall health. Measures of social support yielded little information. CONCLUSIONS: Despite matching for age and body surface area, women did not have as favorable an outcome after surgery as men. Continued research needs to further examine the interaction of physical outcomes and depression in women after coronary artery bypass surgery. PMID- 14528189 TI - Gender differences in the health related quality of life of older adults with heart failure. AB - OBJECTIVE: The objective of this study was to explore whether gender differences exist in health-related quality of life for older adult men and women with heart failure. DESIGN: A secondary data analysis was conducted on an existing dataset of heart failure patients. The original study used an exploratory, correlational longitudinal design. SUBJECTS: 138 subjects (69 men and 69 women) with heart failure completed both data points of the study. RESULTS: Men and women were similar on the number and types of heart failure-related symptoms they experienced. Twenty-seven percent of the subjects could be classified as depressed. The men had better physical functioning than the women did with gender making a greater contribution to physical functioning than age or marital status. Twenty-three percent of the subjects were rehospitalized 4 to 6 weeks after the initial hospital discharge. CONCLUSIONS: This study showed that the health related quality of life for older men and women with heart failure is significantly impacted by heart failure. Older men and women have similar heart failure symptoms, depression, and rehospitalization following a hospital admission for heart failure. Both older men and older women with heart failure have impairments in their physical functioning with older women more physically impaired than older men. PMID- 14528190 TI - The lived experience of survivors of prolonged mechanical ventilation: a phenomenological study. AB - BACKGROUND: Successful liberation from prolonged mechanical ventilation (PMV) is a challenging phenomenon. Whereas many physiological factors have been linked to successful PMV liberation, the psychosocial components are not well delineated. OBJECTIVES: This article serves to describe the experience of patients who survived PMV and to identify salient factors that contributed to successful liberation. METHODS: A phenomenological approach was used to explore the lived experience of seven PMV survivors. RESULTS: Six mutually exclusive themes emerged from the participants' descriptions to create a structural description of the lived experience. CONCLUSIONS: Survivors credited their own self-determination and the expertise and care of health care professionals with their ability to be successfully liberated from prolonged mechanical ventilation. Although surviving PMV was described as frightening and traumatic, comfort and resolve were derived from family members, religion, prayer, and angelic encounters. These findings are useful in providing direction for critical care clinical practice and future research. PMID- 14528191 TI - Are self-reports of breathing effort and breathing distress stable and valid measures among persons with asthma, persons with COPD, and healthy persons? AB - BACKGROUND: Breathing is a subjective experience that includes physical sensations, such as effort to breathe, and an affective element, such as breathing distress. OBJECTIVE: The overall purpose of this investigation was to evaluate whether measurement of the physical sensations (breathing effort) and affective response to these sensations of breathing (breathing distress) are consistent and valid. DESIGN: A longitudinal repeated measures design was used to evaluate a 2-week daily breathing with a sub-sample (n = 43) who also recorded their daily breathing during 4 weeks. SUBJECTS: Age-matched, stable subjects (n = 92) with an average age of 62 were evaluated. The sample consisted of 32 subjects with chronic obstructive pulmonary disease, 27 subjects with asthma, and 33 healthy subjects. MEASURES: Visual Analogue Scales for breathing effort (VAS-E) and breathing distress (VAS-D) were scored daily. RESULTS: The VAS-E and VAS-D mean, highest, and lowest scores were found to be stable over time in the sub sample and a significant difference (F = 2.56, P <.05) between VAS-E and VAS-D was found. Differences were found in mean and highest VAS-E and VAS-D by group, with the COPD group reporting the highest values. CONCLUSIONS: This investigation provided initial evidence of the stability and validity of daily VAS-E and VAS-D measures and preliminary support for the use of daily VAS logs to evaluate differences in breathing effort and breathing distress. PMID- 14528192 TI - Tachycardia in Ebstein's anomaly. PMID- 14528193 TI - Management of anxiety after acute myocardial infarction. PMID- 14528201 TI - Guidelines for conscious sedation and monitoring during gastrointestinal endoscopy. AB - This is one of a series of statements discussing the utilization of GI endoscopy in common clinical situations. The Standards of Practice Committee of the American Society for Gastrointestinal Endoscopy prepared this text. In preparing this guideline, a MEDLINE literature search was performed, and additional references were obtained from the bibliographies of the identified articles and from recommendations of expert consultants. When little or no data exist from well-designed prospective trials, emphasis is given to results from large series and reports from recognized experts. Guidelines for appropriate utilization of endoscopy are based on a critical review of the available data and expert consensus. Further controlled clinical studies are needed to clarify aspects of this statement, and revision may be necessary as new data appear. Clinical consideration may justify a course of action at variance to these recommendations. PMID- 14528202 TI - Monopolar coagulation versus conventional endoscopic treatment for high-risk peptic ulcer bleeding: a prospective, randomized study. AB - BACKGROUND: Severe upper GI hemorrhage from an actively bleeding peptic ulcer is sometimes difficult to treat by conventional endoscopic means, such as multipolar electrocoagulation, heat probe coagulation, and injection therapy. It was hypothesized that monopolar coagulation with a "hot biopsy" forceps may be more effective in such cases. METHODS: A prospective, randomized trial was performed to assess the safety and short-term efficacy of monopolar coagulation versus conventional treatment (combination heat probe coagulation and injection therapy) in the treatment of patients with various types of actively bleeding or high-risk ulcers. RESULTS: Primary hemostasis was achieved in 54 of 56 actively bleeding patients in the monopolar coagulation group. In contrast, primary hemostasis was achieved in 33 of 43 patients in the control group (p = 0.003), with 8 of the remaining 10 being treated successfully by crossover monopolar coagulation. During 30-day follow-up, bleeding recurred in 6 patients in the conventional therapy group versus none in the monopolar coagulation group (p = 0.012). Post endoscopy blood transfusion requirements were lower in the monopolar coagulation group. Mortality and surgery rates were zero in both groups. There was no procedure-related complication. CONCLUSIONS: This study demonstrated improved primary hemostasis and a reduced short-term rate of recurrent bleeding for patients with actively bleeding and high-risk non-bleeding peptic ulcers treated by monopolar coagulation. The complication rate associated with monopolar coagulation was low. PMID- 14528203 TI - Acute small bowel bleeding: a distinct entity with significantly different economic implications compared with GI bleeding from other locations. AB - BACKGROUND: Historically, acute lower intestinal bleeding has incorporated small bowel with colonic sources. This potentially obscures the unique characteristics of small bowel bleeding, which are eclipsed by the attributes of the much more common colonic bleeding. Separating acute lower intestinal bleeding into small bowel and colonic sources may delineate characteristics of each, thereby making it possible to determine whether clinical outcomes vary by anatomic level of bleeding. METHODS: A total of 29 consecutive patients (15 women, 14 men; age 68.6 +/-2.4 years) with acute small bowel bleeding were compared with two other groups, each with 29 consecutive patients, with either acute colonic bleeding or acute upper GI bleeding. Clinical presentation, outcomes, and resource utilization for small bowel bleeding were compared with similar parameters for acute colonic bleeding and upper GI bleeding. RESULTS: Although the clinical presentation did not always distinguish the 3 groups, resource utilization was significantly higher in the small bowel bleeding group. The latter group required a higher number of diagnostic procedures (p < 0.001) and blood transfusions (p < 0.001), remained in hospital longer (p < 0.05), and had a higher cost of hospitalization (p < 0.001) compared with the colonic bleeding and upper GI bleeding groups. The mortality rate for patients with small bowel bleeding was 10%. Although none of the patients with upper GI bleeding and only 14% of those with colonic bleeding required greater than 3 diagnostic procedures, 79% of patients with small bowel bleeding required 4 procedures for diagnostic localization (p < 0.0001). CONCLUSIONS: Small bowel bleeding ("mid-intestinal bleeding") is a distinct clinical entity with significantly worse outcomes compared with colonic bleeding and upper GI bleeding. The focus of the investigation should be directed to the small bowel, with enteroscopy or capsule endoscopy, when 3 investigative procedures fail to localize recurrent overt GI bleeding. PMID- 14528204 TI - Prognostic factors for recurrence of bile duct stones after endoscopic treatment by sphincter dilation. AB - BACKGROUND: The long-term outcome for patients after endoscopic sphincter of Oddi dilation is poorly documented. This study investigates the recurrence rate for bile duct stones in patients followed for 1 year or more after endoscopic sphincter dilation and stone extraction, and assessed prognostic factors associated with recurrence of ductal calculi. METHODS: A total of 169 patients with bile duct stones were treated with endoscopic sphincter dilation between July 1998 and August 2001. Follow-up studies consisted of periodic biochemical tests and out-patient evaluations with endoscopic retrograde cholangiography or magnetic resonance cholangiography performed when follow-up exceeded 1 year. Putative risk factors for stone recurrence included gender, age, stone size and number, associated peripapillary diverticulum, gallbladder status, color of bile duct stones, and bile duct diameter. Statistical analysis consisted of both a Kaplan-Meier estimation and a multivariate Cox regression model. RESULTS: Complete stone clearance was achieved in 162 (95.8%) patients, of whom 151 were followed (13 patients died from unrelated disorders). Mean follow-up was 23 months. Stone recurrence was documented in 13 patients. Patients with dilated bile duct or peripapillary diverticulum were at high risk for recurrence. CONCLUSIONS: The interval between treatment of bile duct stones by endoscopic sphincter dilation and the recurrence of biliary calculi is relatively short. Bile duct size and peripapillary diverticula are risk factors for early recurrence. PMID- 14528205 TI - Determination of sphincter of Oddi dysfunction in patients with prior normal manometry. AB - BACKGROUND: Assessment of sphincter of Oddi motility by manometry is limited to a finite time period, and the presence of a motor disorder that is intermittent may not be documented. This study evaluated the frequency of sphincter of Oddi dysfunction in persistently symptomatic patients with previously normal sphincter of Oddi manometry studies. METHODS: A total of 177 patients underwent ERCP for suspected sphincter of Oddi dysfunction and had a normal sphincter of Oddi manometry study (both biliary and pancreatic) over a 5-year period (1996-2001). All patients referred for a second ERCP with sphincter of Oddi manometry for evaluation of persistent symptoms were included in this study. RESULTS: Of the 177 patients, 12 (mean age 37.6 years, range 19-59 years) met criteria for inclusion. The mean time interval between the first and second ERCP was 337 days (range 43-792 days). Sphincter of Oddi dysfunction was diagnosed at a second sphincter of Oddi manometry in 5 (42%) patients; 4 had pancreatic sphincter hypertension; one had elevation of both pancreatic and biliary sphincter pressures. All 5 patients underwent endoscopic sphincter ablation therapy; 4 were symptom-free on follow-up at, respectively, 26, 40, 48, and 72 months; one patient had persistent symptoms from pancreatic sphincter restenosis and required multiple endoscopic interventions. Five of the 12 (42%) patients with normal manometric studies were found to have pancreatographic changes of chronic pancreatitis; the two remaining patients had a normal ERCP and manometry. CONCLUSIONS: A single negative manometry study does not rule out sphincter of Oddi dysfunction. Repeat ERCP with manometry may be warranted for patients with persistent symptoms in whom the clinical suspicion for sphincter of Oddi dysfunction remains high. PMID- 14528206 TI - Perceptions of gastroenterology fellows regarding ERCP competency and training. AB - BACKGROUND: The adequacy of ERCP training in the United States may be suboptimal because many training programs do not provide fellows with the exposure to the procedures necessary to achieve competence. METHODS: A short survey questionnaire, which assesses the training program, the personal ERCP experience, the perceptions regarding training adequacy, and the post-training practice plans, was sent to all fellows graduating from gastroenterology training programs. RESULTS: Graduating fellows performed a median of 140 ERCPs and 35 sphincterotomies during training, with an associated median comfort level for independently performing sphincterotomy of 7.5 on a scale of 1 to 10. The median estimated success rate for independent free cannulation was 75%. Based on nonparametric correlation and regression analysis, 180 ERCPs would be necessary to achieve a free cannulation rate of 80% and 69 sphincterotomies to achieve a comfort level of 8 on a scale of 1 to 10. Thirty-six percent of fellows achieved the number of procedures and cannulation success determined by this study to indicate procedural competence. Sixty-four percent of fellows did not achieve procedural competence and 33% reported inadequate ERCP training. Nevertheless, 91% of fellows expected to perform unsupervised ERCP after training. CONCLUSIONS: The results of this study are consistent with those of previously published studies demonstrating that 160 to 200 ERCP procedures are necessary to achieve competence to perform ERCP. The majority of graduating fellows do not achieve an acceptable success rate during training, yet still intend to perform ERCP after training. PMID- 14528207 TI - A predictive model for length of Barrett's esophagus with hiatal hernia length and duration of esophageal acid exposure. AB - BACKGROUND: A significant correlation between the duration and height of esophageal acid exposure and the length of Barrett's mucosa has been demonstrated. The aims of this study were to determine if there is a correlation between hiatal hernia length and Barrett's esophagus length, and to develop a predictive model for Barrett's esophagus length by using hiatal hernia length and duration of esophageal acid exposure. METHODS: Consecutive patients with Barrett's esophagus diagnosed endoscopically were enrolled in the study. Barrett's esophagus was defined by the presence of intestinal metaplasia in biopsy specimens obtained from salmon-colored mucosa extending into the esophagus. Barrett's mucosa 3 cm or greater in length was considered long-segment Barrett's esophagus; and less than 3 cm long was considered short-segment Barrett's esophagus. Hiatal hernia was considered present if the esophagogastric junction was displaced 1 cm or more proximal to the diaphragmatic hiatus. RESULTS: Twenty-four men (mean age 66.1 +/-2.4 [SE]) with Barrett's esophagus were included in this study. Mean Barrett's length was 4.1 +/-0.7 cm. The Pearson correlation coefficient between hiatal hernia length and Barrett's esophagus length was 0.62 (p < 0.01). Similarly, there was a significant correlation between esophageal acid exposure and Barrett's length (r = 0.62; p < 0.01). Multiple linear regression analysis revealed that hiatal hernia length and duration of esophageal acid exposure were associated significantly with length of Barrett's mucosa (R(2) = 0.54; p < 0.001). A regression equation was developed expressing mean Barrett's length (cm) = 0.79 + (0.68) hernia length (cm) + (0.075) duration of esophageal acid exposure (% time pH < 4). CONCLUSIONS: The length of Barrett's mucosa correlated with the length of hiatal hernia. A predictive model for Barrett's length by using hiatal hernia length and duration of esophageal acid exposure was developed. This suggested that these two pathophysiologic factors are good predictors of the length of Barrett's mucosa. PMID- 14528208 TI - Use of large-diameter metallic stents to seal traumatic nonmalignant perforations of the esophagus. AB - BACKGROUND: Surgery for traumatic, non-malignant perforation of the esophagus in patients presenting more than 24 hours after its occurrence carries a high morbidity and mortality. Covered metallic stents have been used to effectively seal perforations in individual patients with Boerhaave's syndrome. METHODS: Eleven consecutive patients presented with esophageal perforation that was caused by Boerhaave's syndrome (n = 5), resection of an epiphrenic diverticulum (n = 2), rigid esophagoscopy (n = 2), extended gastric resection (n = 1), or pneumatic dilation for achalasia (n = 1). A large diameter Flamingo Wallstent (proximal/distal diameters, 30/20 mm) (7 patients) or a large diameter Ultraflex stent (proximal/distal diameters, 28/23 mm) (4 patients) was placed. Pleural cavities were drained with thoracostomy drains, and antibiotics were administered. RESULTS: The median time from perforation to stent insertion was 60 hours (range, 24 hours to 28 days). The perforation was totally sealed in 10 of 11 patients. Two patients underwent esophageal resection because of incomplete sealing of the perforation or incomplete drainage of the pleural cavity and mediastinum. The other 9 patients recovered uneventfully and resumed a normal diet within 7 to 18 days. In 7 patients, the stents were retrieved endoscopically after a median of 7 weeks (range, 6 to 14 weeks), whereas two patients refused to have the stent retrieved (in one, the stent migrated into the stomach; the other patient died 6 months after stent placement from an unrelated cause). CONCLUSIONS: Traumatic perforation of the esophagus can be treated successfully with large diameter metallic stents, together with adequate drainage of the thoracic cavity. PMID- 14528209 TI - Diagnosis of benign cysts of the mediastinum: the role and risks of EUS and FNA. AB - BACKGROUND: Benign mediastinal cysts, which account for approximately 20% of mediastinal masses, may be diagnostic challenges. Information regarding the use of EUS and EUS-guided FNA in this setting is limited. The aim of this study was to demonstrate the value and potential risks of EUS and EUS-FNA in the diagnosis of mediastinal foregut cysts. METHODS: The EUS database of a single tertiary referral center was reviewed for the diagnosis of benign mediastinal cysts. Twenty patients were identified who underwent 23 EUS examinations for suspected mediastinal cysts (n = 4), for follow-up of a known cyst (n = 3), or for a mediastinal mass of unknown origin (n = 16). RESULTS: In 19 patients, the definite diagnosis of a mediastinal cyst was established by EUS. Twelve cysts appeared anechoic, 6 were hypoechoic, and one anechoic cyst contained small echoic foci. CT (n = 17) or magnetic resonance imaging (n = 1) was performed in 18 cases; only 4 of these were diagnostic of a cyst. In 3 cases, the cyst contents were aspirated by EUS-FNA. In a fourth case, a solid-appearing duplication cyst, misdiagnosed by EUS, was sampled with FNA and core biopsy. This patient developed severe sepsis secondary to mediastinitis 4 days later. Thoracotomy revealed an infected bronchogenic cyst. CONCLUSIONS: EUS provides a minimally invasive approach to the diagnosis of benign mediastinal cysts and may be more accurate than CT or other imaging modalities. Aspiration of suspected cysts should be undertaken with caution, given the risk of infection. PMID- 14528210 TI - Acute GI bleeding in the setting of supratherapeutic international normalized ratio in patients taking warfarin: endoscopic diagnosis, clinical management, and outcomes. AB - BACKGROUND: Acute GI bleeding is a life-threatening complication of warfarin therapy. Acute GI bleeding in patients with an international normalized ratio of 4.0 or greater (supratherapeutic) is often attributed to trivial mucosal lesions. The aim of the study was to determine the frequency of potentially significant lesions that would warrant endoscopy in this setting. METHODS: A retrospective review was conducted of patients treated with warfarin who were admitted to a single Veterans Affairs hospital from 1996 to 2000 with acute GI bleeding. Endoscopic findings, clinical management, and outcomes are reviewed for patients with a supratherapeutic international normalized ratio (>or=4.0) and compared with patients with an international normalized ratio in the therapeutic range (2.0-3.9). RESULTS: Fifty-five patients with an international normalized ratio of 4.0 or greater (mean 8.4 [3.9]) and 43 patients with an international normalized ratio between 2.0 and 3.9 (mean 2.9 [0.6]) were hospitalized with acute GI bleeding. Thirty-seven patients (67%) with a supratherapeutic international normalized ratio and GI bleeding underwent upper endoscopy. Of these, 81.1% had positive findings, 18.9% had peptic ulcer disease, and 7.2% required endoscopic treatment. Thirty-eight percent of the patients with a supratherapeutic international normalized ratio underwent lower endoscopy; of these, 57.1% had abnormal findings and 9.5% required endoscopic treatment. Four patients (7.3%) in the supratherapeutic international normalized ratio group died during the index hospitalization. When patients with GI bleeding and a therapeutic international normalized ratio were compared with those with a supratherapeutic international normalized ratio, there were no significant differences between the two groups with regard to days of hospitalization, units of blood transfused, frequency of recurrent bleeding, need for surgery, or in-hospital deaths. CONCLUSIONS: The high frequency of clinically significant lesions in patients taking warfarin with an international normalized ratio in the supratherapeutic range and acute GI bleeding supports a role for endoscopic evaluation. PMID- 14528211 TI - Long-term outcomes for patients with post-liver transplant anastomotic biliary strictures treated by endoscopic stent placement. AB - BACKGROUND: Biliary stricture is one of the most common complications of liver transplantation. A number of treatment options are available, but a standard approach has not been established. METHODS: A total of 25 patients with post liver transplantation anastomotic strictures were treated endoscopically by stent placement. Long-term outcomes (bile duct patency, morbidity, and mortality) were reviewed retrospectively. RESULTS: Placement of a stent was attempted in 25 patients with anastomotic stricture. Successful stent placement with stricture resolution at the time of stent removal was noted in 22 patients (technical success 88%). In those 22 patients, long-term success (mean follow-up after all stents removed, 54 months) was observed in 20 patients (90%) and partial success in two (10%). Long term, failure did not occur in any patient. There was no procedure- or disease-related mortally. Three mild episodes of cholangitis occurred during the period while the stents were in place, in relation to 79 endoscopic interventions for a procedure-related complication rate of 3.7%. CONCLUSIONS: The long-term outcome for patients with post-liver transplantation biliary anastomotic strictures treated with endoscopic stent placement is excellent, with no therapy- or disease-associated mortality and minimal morbidity. PMID- 14528212 TI - Pretreatment laparoscopic appearance of the liver can predict response to combination therapy with interferon alpha 2B and ribavirin in chronic hepatitis C. AB - BACKGROUND: The aim of this study was to determine whether the pretreatment laparoscopic appearance of the liver is an additional predictor of response to combination therapy with interferon and ribavirin in patients with hepatitis C. METHODS: A retrospective review was performed of 112 patients (61 men, 51 women [ratio 1.3:1]; mean age 50 [10] years, range 15-73 years) with untreated hepatitis C, without other causes of liver disease, who underwent diagnostic laparoscopy before combination therapy with interferon and ribavirin for at least 24 weeks with a 24-week post-treatment follow-up. Fifty-nine were white, 37 Hispanic, and 16 African American. Patients were divided into responders and non responders based on viral clearance. Demographics, genotype, pre-therapy hepatitis C virus RNA, histopathologic, and laparoscopic appearances were analyzed. RESULTS: Ninety-three patients (83%) had genotype 1 with a mean pretreatment hepatitis C virus RNA of 3.2 (2.8) million copies/mL. Thirty-seven (33%) had laparoscopic evidence of cirrhosis, whereas, only 30 (26.4%) had cirrhosis by histopathologic criteria. Patients were treated with interferon and ribavirin (mean dose 10.6 [2.5] mg/kg) for a mean duration of 37.7 (11.4) weeks, depending on response and genotype. A sustained response was observed in 26 (23%) patients; in 12 (11%), there was only a biochemical response (biochemical responder), while 59 (53%) and 15 (13%) were classified, respectively, as non responders and relapsers. Logistic regression analysis revealed that pretreatment laparoscopic appearance (p = 0.034) and genotype (p = 0.002) were significant predictive factors; that is, a lesser extent of fibrosis at laparoscopy and genotypes other than 1 were predictive of a sustained response to combination therapy. CONCLUSIONS: Pretreatment laparoscopic appearance alone and genotype are significant predictors of a sustained response to combination therapy in patients with hepatitis C. Laparoscopy and biopsy are complementary for the diagnosis of cirrhosis in hepatitis C. PMID- 14528213 TI - Diagnostic and therapeutic ERCP in hepatic hydatid disease. AB - BACKGROUND: This retrospective study evaluated the use of diagnostic and therapeutic ERCP in pre- and postoperative patients with hepatic hydatid disease. METHODS: For 8 years, ERCP was performed in 39 patients with hepatic echinococcal disease. Indications in the preoperative group of patients (n = 19) included a cholestatic enzyme profile in all cases; jaundice or acute cholangitis also was present in, respectively, 14 and 7 cases. In the postoperative group (n = 20), indications for ERCP included persistent external biliary fistula after surgery in 10 patients, jaundice in 8, acute cholangitis in 7, and right upper quadrant pain in 2 patients. RESULTS: ERCP findings in the preoperative group included cystobiliary fistula (9 patients), external compression of the hepatic biliary system (5), hydatid vesicles and/or membranes within the biliary tract (3), intrahepatic duct stricture (1), and a normal cholangiogram (4). The most common ERCP finding in the postoperative group was external biliary fistula (10 patients); other findings consisted of hydatid cyst material within the bile duct (4), bile duct stenosis (2), cystobiliary fistula and hydatid cyst material in the bile duct (1), cystobiliary fistula (1), hydatid membranes in the gallbladder (1), extrinsic compression to bile ducts (1), and a normal cholangiogram (1). In the preoperative group, endoscopic sphincterotomy was performed in 11 patients, with balloon catheter extraction in 2; complete resolution of findings was achieved in 10 cases. In the postoperative group, sphincterotomy (with balloon or basket extraction as needed) was performed in 19 patients, stents were placed in 2 patients, 1 patient underwent balloon dilatation, and 1 had nasobiliary drainage; there was complete resolution of the findings in 14 of the 20 patients. CONCLUSIONS: ERCP and related therapeutic maneuvers are safe and valuable in the pre- and postoperative management of patients with hepatic hydatid disease. PMID- 14528215 TI - Acute GI bleeding: upper and lower, but is there a "middle kingdom"? PMID- 14528214 TI - Diagnosis and management of hereditary non-polyposis colon cancer. PMID- 14528216 TI - "Trained in ERCP". PMID- 14528217 TI - Pancreatic duct-portal vein fistula. PMID- 14528218 TI - Gastric herniation through PEG site. PMID- 14528219 TI - Esophageal foreign body. PMID- 14528220 TI - Pneumatosis cystoides intestinalis. PMID- 14528221 TI - Giant jejunal stone on ERCP. PMID- 14528222 TI - Sclerosing cholangitis caused by old shrapnel injury. PMID- 14528223 TI - Enteral self-expandable stents. PMID- 14528224 TI - Mini-laparoscopically guided percutaneous gastrostomy and jejunostomy. AB - BACKGROUND: Percutaneous endoscopic tube placement can be problematic under certain circumstances: absence of transillumination of the abdominal wall, percutaneous jejunostomy in patients with a PEG tube and recurrent aspiration, enteral feeding access after gastrectomy, and obstruction of the upper GI tract. As an alternative in these problematic situations, a technique was developed for placing feeding tubes under visual control by using mini-laparoscopy. METHODS: Placement of a feeding tube with mini-laparoscopy with the patient under conscious sedation was considered for 17 patients in whom standard PEG placement was impossible. Techniques used were the following: combined mini laparoscopy/endoscopy for placement of a percutaneous gastrostomy or jejunostomy, and mini-laparoscopic-guided direct tube placement in cases of obstruction of the upper GI tract. OBSERVATIONS: In 13 patients, mini-laparoscopic-assisted tube placement was successful. In 4 patients, adhesions or peritoneal carcinomatosis prevented laparoscopic visualization of the stomach or small bowel. The combined mini-laparoscopic/endoscopic approach allowed a successful insertion of gastric tubes in 6 patients and jejunal tubes in 4 patients. Direct insertion of a percutaneous endoscopic jejunostomy tube without enteroscopy was feasible in all 3 patients with obstruction of the upper GI tract. No complication occurred. CONCLUSIONS: Mini-laparoscopy-assisted tube placement is a simple and safe alternative when endoscopic percutaneous tube placement is problematic or not feasible. PMID- 14528225 TI - Pancreatic duct stent insertion for functional smoldering pancreatitis. AB - BACKGROUND: Some patients with smoldering pancreatitis in the absence of necrosis, pseudocyst, and ductal disruption experience unremitting abdominal pain caused by persistent pancreatic inflammation. Experience with the use of pancreatic duct stents in this patient population was reviewed. PATIENTS AND METHODS: Data for 11 patients with smoldering pancreatitis who underwent ERCP with pancreatic duct stent placement were reviewed retrospectively. All patients had severe, daily pain that worsened with ingestion of food, had required narcotic analgesics for control of pain, had lost weight, and had persistently elevated serum levels of pancreatic enzymes as well as pancreatic inflammatory changes on CT (without necrosis or pseudocyst). Six patients were being treated with parenteral nutrition. OBSERVATIONS: The mean duration of symptoms from the onset of pancreatitis until pancreatic duct stent insertion was 74 days (range 14 151 days). Stents were placed for a mean of 7 weeks (range 2-19 weeks). Pancreatic stent placement provided permanent pain relief in 10 (91%) patients within a mean of 9 days (range 3-20 days); one patient had persistent symptoms requiring celiac plexus blockade after 5 months. Parenteral nutrition and treatment with narcotic agents were discontinued for 10 patients within a mean of 15 days (range 7-39 days) after pancreatic duct insertion. CONCLUSIONS: Smoldering pancreatitis may result from functional obstruction, possibly caused by edema or spasm, of the papillary orifice. Insertion of a stent into the pancreatic duct alleviates pain, enables early resumption of oral intake of food, and facilitates pancreatic duct drainage. It also may help to prevent complications arising from persistent pancreatic inflammation. PMID- 14528226 TI - Malacoplakia of the stomach: case report and review. PMID- 14528227 TI - Bile duct obstruction caused by an eosinophilic papillary pseudotumor. PMID- 14528228 TI - Colonic actinomycosis presenting as diffuse colitis and protein-losing enteropathy: a case report. PMID- 14528229 TI - Development of specialized columnar epithelium in the esophagus of two patients with esophageal strictures caused by corrosive ingestion. PMID- 14528230 TI - Successful treatment of esophagojejunal disconnection after total gastrectomy by insertion of a covered self-expandable esophageal metallic stent. PMID- 14528231 TI - EUS-guided trucut biopsy for diagnosis of an esophageal stromal tumor: case report. PMID- 14528232 TI - Intraoperative transjejunal ERCP: case reports. PMID- 14528233 TI - Endoscopic microwave lithotripsy of a biliary stone lodged at a choledochoduodenal anastomosis. PMID- 14528234 TI - Endoscopic incision for esophageal intramural hematoma after injection sclerotherapy: case report. PMID- 14528235 TI - Large colonic lipoma with mucosal ulceration mimicking carcinoma. PMID- 14528236 TI - Safety of sodium phosphate for colonoscopy. PMID- 14528238 TI - Botulinum toxin for spastic GI disorders. PMID- 14528239 TI - Ligate and let go. PMID- 14528241 TI - Scanning laser ophthalmoscope-evoked multifocal ERG (SLO-mfERG) in patients with macular holes and normal individuals. AB - AIMS: A scanning laser ophthalmoscope (SLO) has been used for multifocal electroretinography (mf ERG) measurements under simultaneous fundus monitoring. The aim of this study was to prove if the SLO-mfERG measurement reflects reliably the clinically registered underlying disease, and to demonstrate the importance of its main advantage, fixation monitoring. METHODS: In all, 10 patients with macular hole stage II/III were included in the study, and 19 normal individuals served as the control group. The mf ERG device was combined with an SLO, which was used both as a stimulus and trigger unit as well as a fundus-monitoring system. Monitoring of the fundus was guaranteed by an infrared laser (780 nm). The stimulus matrix consisted of 61 hexagonal elements, covering 24 degrees of the posterior pole. We examined both, patients with macular holes and healthy individuals. RESULTS: Compared to normal controls, patients with a macular hole (Gass stage III) showed a significant decrease in response density in the centre of the stimulus array, which correlated well with the morphological alteration observed by clinical examination. However, variation of response density of the central hexagonal area has been proved to be high. CONCLUSIONS: SLO-mfERG is a feasible and reliable new technique to investigate macular function under simultaneous fundus control. The main advantage is that control of fixation can be used in order to obtain more reliable results that correlate well with visible fundus abnormalities such as in patients with macular holes. However, further investigations have to be performed in order to overcome sufficiently the problem of fixation instability. PMID- 14528242 TI - Clinical features associated with survival of patients with lymphoma of the ocular adnexa. AB - PURPOSE: Although systemic or eyelid involvement by ocular adnexal lymphoma carries a worse prognosis, there have been few reports of the outcome in relation to clinical presentation. The outcome of malignant ocular adnexal lymphoma was, therefore, related to presenting clinical symptoms and signs. DESIGN AND METHODS: A retrospective, noncomparative case-note review of 326 patients treated in the Orbital Clinic at Moorfields Eye Hospital. The associations between presenting symptoms or signs and three outcome measures (v.i.) were assessed by univariate and multiple variable regression together with Kaplan-Meier analysis. MAIN OUTCOME MEASURES: (i) Presence of extra-orbital disease at the time of presentation; (ii) development of systemic lymphoma after new presentation with solely ocular adnexal disease; and (iii) death attributable to widespread lymphoma. RESULTS: Presentation with disseminated disease was rarer with over 1 year's ophthalmic symptoms (odds ratio (OR) 0.7; 95% CI 0.5-0.9) and much more frequent with bilateral adnexal disease (OR 5.8; 95% CI 3.0-11.2). With solely adnexal disease at presentation, subsequent extra-orbital lymphoma was more frequent and earlier with lacrimal gland disease (as compared to those without; hazard ratio (HR) 1.9; 95% CI 1.2-4.5) or with eyelid disease (compared to those without; HR 2.4; 95% CI 1.2-4.5), or with bilateral disease (compared to unilateral disease; HR 2.6; 95% CI 1.4-5.2). Prior or concurrent systemic disease was the most significant predictive factor for lymphoma-related death (HR 6.8; 95% CI 4.3-10.9), but tumour-related death was also commoner and earlier with bilateral disease (HR 2.4; 95% CI 1.4-4.0) or where a relative afferent papillary defect was present (HR 2.8; 95% CI 1.6-4.9). Similarly, the rate of tumour related death was slightly less where symptoms had been present for more than a year (HR 0.8; 95% CI 0.7-1.0) and slightly greater in the elderly (HR 1.03; 95% CI 1.01-1.05). Conjunctival lymphoma had the lowest rate of extra-orbital spread and lymphoma-related death, the rate of these two events being sequentially greater for patients with predominantly deep orbital lymphoma, lacrimal gland lymphoma, or eyelid lymphoma. CONCLUSION: These data suggest that presenting symptoms and signs of patients with ocular adnexal lymphoma are significantly associated with the risk of systemic disease at orbital presentation, the rate of subsequent spread, and the rate of lymphoma-related death. PMID- 14528243 TI - Ocular rubber bullet injuries. AB - PURPOSE: To report the type and severity of ocular and orbital injuries caused by rubber bullets. METHODS: A total of 42 consecutive patients seen over a 3-month period with ocular and orbital rubber bullet injuries were assessed clinically and radiographically within 1 day of injury, and the findings were recorded. Clinical outcomes following treatment were also recorded up to 6 months postinjury. RESULTS: A total of 90% of the patients were male. The mean age of patients was 25.0 years (4-60). Of the patients, 54% had lid or skin lacerations, 40% hyphaema, 38% ruptured globe, 33% orbital fracture, 26% retinal damage, and 21% retained rubber bullet in or around the orbit. At follow-up, 53% of the patients had a visual acuity of less than 6/60, 7% less than 6/18 to 6/60, and 40% 6/18 or better. CONCLUSIONS: The term 'rubber bullet' is misleading. 'Rubber bullets' cause a wide variety of ocular and periocular injuries. Orbital fractures are common. The tissues of the orbit are easily penetrated. If the globe is hit, it is rarely salvageable. PMID- 14528244 TI - Repeatability and reproducibility of the BVI ultrasonic Pachymeter. AB - PURPOSE: To evaluate the interobserver and intraobserver reliability of a commercially available, portable, ultrasonic pachymeter. METHODS: For the interobserver variability study, 42 healthy subjects underwent repeated ultrasonic pachymetry under topical anaesthesia performed by two observers. For the intraobserver study, another 30 further healthy subjects underwent repeated pachymetry by one of the observers. Agreement was analysed by means of Bland Altman plots and by determination of the intraclass correlation coefficient. RESULTS: For the interobserver variability study, the mean measurement difference between observers was 0.7 microm (95% CI -0.8-2.2 microm) and the intraclass correlation coefficient was 0.9958 (95% CI 0.9922-0.9977). The Bland-Altman plot showed narrow limits of agreement with respect to central corneal thickness (CCT). For the intraobserver variability study, the mean difference between the repeated measurements was 0.9 microm (95% CI -3.1-1.3 microm). The intraclass correlation coefficient was 0.9934 (95% CI 0.9863-0.9969). A Bland-Altman plot again showed narrow limits of agreement with respect to CCT. The mean CCT for 72 subjects was 538 microm (95% CI 528-545 microm). CONCLUSIONS: Measurements of CCT using the BVI Pocket Pachymeter were repeatable and had excellent interobserver reliability. Measurement variation amounted to less than 0.2% assuming a mean CCT of 538 microm. PMID- 14528246 TI - Ciliary effusion complicates blunt ocular trauma. PMID- 14528245 TI - Intraoperative 5-fluorouracil application during primary trabeculectomy in Nigeria: a comparative study. AB - PURPOSE: To investigate the effect on intraocular pressure (IOP) of intraoperative 5-fluorouracil (5-FU) application during primary trabeculectomy in a West African population. METHODS: Retrospective case note search of operating theatre records for primary trabeculectomy between 1996 and 2000 in an eye hospital in Lagos, Nigeria. For statistical analysis, patients were divided into group that received intraoperative 5-FU (50 mg/ml for 5 min) and a control group that received no intraoperative antimetabolites. Patients with a follow-up of less than 6 months were excluded. For life-table analysis, only one eye from each patient was included to maintain independent analysis. RESULTS: A total of 154 eyes of 101 patients were included in the study. Average follow-up was 17 +/- 2.18 (95% confidence interval) months. No significant difference was found between the groups for age, sex, preoperative medication, and follow-ups. Comparison of survival curves by the log-rank test showed that the probability of maintaining an IOP of 20 mm Hg or less at 18 months was 76% in the 5-FU group and 79% in the controls (P= 0.55). However, the probability of maintaining an IOP 14 mmHg or less was significantly greater in the 5-FU group (64%) than in the controls (39%, P=0.018). Visual acuity loss of more than two Snellen-chart lines was observed in six eyes (7.9%) of the 5-FU group and in four eyes (5.1%) of the control group and this difference was not significant (P=0.49, Fisher exact test). Complication rates were similar in each group. CONCLUSIONS: In this population, intraoperative 5-FU application during trabeculectomy appears to improve probability of maintaining an intraocular pressure of 14 mmHg or less with no additional deleterious effects on the eye. PMID- 14528247 TI - Limbal stem cell deficiency: a clinical chameleon. PMID- 14528248 TI - Accelerated growth of a primary orbital schwannoma during pregnancy. PMID- 14528249 TI - Scedosporium apiospermum keratomycosis with secondary endophthalmitis. PMID- 14528250 TI - Primary ductal adenocarcinoma of the lacrimal gland in a patient with neurofibromatosis. PMID- 14528251 TI - Confocal microscopy in bee sting corneal injury. PMID- 14528254 TI - Beyond the horizon. PMID- 14528255 TI - Virtual laboratories in the life sciences. A new blueprint for reorganizing research at the European level. PMID- 14528256 TI - Wish fulfillment and its discontents. PMID- 14528257 TI - Fierce creatures. Zoonoses, diseases that jump from animals to humans, are a growing health problem around the world. Understanding their causes and their effects on humans have therefore become an important topic for global public health. PMID- 14528258 TI - What doesn't kill you makes you stronger. A new model for risk assessment may not only revolutionize the field of toxicology, but also have vast implications for risk assessment. PMID- 14528259 TI - Unhealthy competition. Advances in biomedical research provide new drugs and technologies in medicine, but also new ways for athletes to illegally enhance their performance. PMID- 14528262 TI - Subtleties among subtilases. The structural biology of Kex2 and furin-related prohormone convertases. PMID- 14528263 TI - The ribosome and YidC. New insights into the biogenesis of Escherichia coli inner membrane proteins. AB - In the bacterium Escherichia coli, inner membrane proteins (IMPs) are generally targeted through the signal recognition particle pathway to the Sec translocon, which is capable of both linear transport into the periplasm and lateral transport into the lipid bilayer. Lateral transport seems to be assisted by the IMP YidC. In this article, we discuss recent observations that point to a key role for the ribosome in IMP targeting and to the diverse roles of YidC in IMP assembly. PMID- 14528264 TI - Regulating histone acetyltransferases and deacetylases. AB - Histone acetyltransferases and histone deacetylases regulate the acetylation of histones and transcription factors, and in doing so have major roles in the control of cell fate. Many recent results have indicated that their function is strictly regulated in cells through the modulation of their levels, activity and availability for interaction with specific transcription factors. In this review, we present the various molecular mechanisms that bring about this tight regulation and discuss how these regulatory events influence cellular responses to environmental changes. PMID- 14528265 TI - Finding the right organelle. Targeting signals in mitochondrial outer-membrane proteins. AB - The mitochondrial outer membrane contains a diverse set of proteins that includes enzymes, components of the preprotein translocation machinery, pore-forming proteins, regulators of programmed cell death, and those that control the morphology of the organelle. All these proteins, like the vast majority of mitochondrial proteins, are encoded in the nucleus, so they are synthesized in the cytosol and contain signals that are essential for their subsequent import into mitochondria. This review summarizes our current knowledge of the signals that target mitochondrial outer-membrane proteins to their correct intracellular location. In addition, the mechanisms by which these signals are decoded by the mitochondria are discussed. PMID- 14528266 TI - Adenovirus protein IX sequesters host-cell promyelocytic leukaemia protein and contributes to efficient viral proliferation. AB - The product of adenovirus type 5 (Ad5) gene IX, protein IX (pIX), is a multifunctional protein that stabilizes the viral capsid and has transcriptional activity. We show that pIX also contributes to the Ad5-induced reorganization of the host-cell nuclear ultrastructure: pIX induces the formation of specific and dynamic nuclear inclusions, and the host promyelocytic leukaemia (PML) protein, which is the main structural organizer of PML bodies, is stably relocated and confined within the pIX-induced inclusions late in infection. Our results suggest that Ad5 has evolved a unique strategy that leads to the sustained neutralization of PML bodies throughout infection, thereby ensuring optimal viral proliferation. PMID- 14528267 TI - Exclusion of exogenous phosphatidylinositol-3,4,5-trisphosphate from neutrophil polarizing pseudopodia: stabilization of the uropod and cell polarity. AB - Although there is accumulating evidence that the generation and localization of phosphatidylinositol-3,4,5-trisphosphate (PtdIns(3,4,5)P(3)) have important functions in neutrophil polarization and chemotaxis, the mechanism of this linkage has yet to be established. Here, using exogenous fluorescent PtdIns(3,4,5)P(3) introduced into the inner leaflet of the neutrophil plasma membrane by a cationic carrier, we show that: first, PtdIns(3,4,5)P(3) uniformly delivered to the neutrophil plasma membrane is excluded from newly forming pseudopodia; second, PtdIns(3,4,5)P(3) translocates to and is immobilized at the pole opposite a stable polarizing pseudopod; third, asymmetric delivery of PtdIns(3,4,5)P(3) to the neutrophil triggers the generation of polarizing pseudopodia at the opposite pole; and finally, PtdIns(3,4,5)P(3) triggers repetitive Ca(2+) signals, the onset of which precedes morphological polarization. These data suggest that translocation and immobilization of PtdIns(3,4,5)P(3) or a 3,x-phosphorylated metabolite in the uropod functions as an important polarization cue that defines neutrophil polarity and stabilizes the generation of pseudopodia at the opposite pole. PMID- 14528270 TI - Genetic and epigenetic alterations as hallmarks of the intricate road to cancer. AB - Despite the clonal origin of most tumors, their tremendous heterogeneity suggests that cancer progression springs from the combined forces of both genetic and epigenetic events, which produce variant clonal populations, together with the selective pressures of the microenvironment, which promote growth and, perhaps, dissemination of variants with a specific set of characteristics. Although the importance of genetic mutations in cancer has long been recognized, the role of epigenetic events has been suggested more recently. This review focuses on the genetic and epigenetic molecular mechanisms involved in cancer onset and progression, and discusses the possibility of new strategies in the development of anticancer treatments. PMID- 14528271 TI - Cancer epigenetics. AB - Aberrant DNA methylation of the promoter region is a key mechanism for inactivation of genes that suppress tumorigenesis. Genes that are involved in every step of tumor formation can be silenced by this mechanism. Inhibitors of DNA methylation, such as 5-azadeoxycytidine (5AZA), can reverse this epigenetic event suggesting a potential use in cancer therapy. The structure of chromatin can also play an important role with respect to the regulation of gene expression. Chromatin containing hypoacetylated lysines in histones has a compact structure that is repressive for transcription. Inhibitors of histone deacetylase (HDAC) can convert chromatin to an open structure and activate certain genes that inhibit tumor growth. These HDAC inhibitors also have potential in cancer therapy. A 'cross-talk' between DNA methylation and histone deacetylation can occur and work in concert to silence gene expression. The molecular mechanism involves the attachment of a methylated CpG binding protein (MBP) to the methylated promoters and its recruitment of HDAC to form a complex that suppresses transcription. These two epigenetic modifications represent an interesting target for therapeutic intervention using 5AZA and HDAC inhibitors. These agents in combination have been shown to produce a synergistic reactivation of tumor suppressor genes and an enhanced antineoplastic effect against tumor cells, and should be investigated as a novel form of epigenetic therapy for cancer. PMID- 14528272 TI - Epigenetic targets for immune intervention in human malignancies. AB - Emerging evidences suggest that epigenetic events associated with tumor development and progression, such as deregulated methylation of CpG dinucleotides and aberrant histone acetylation, may impair the immunogenic potential of cancer cells. In fact, DNA hypermethylation and/or histone deacetylation contribute to the absent or downregulated expression of different components of the 'tumor recognition complex' (i.e., HLA class I antigens, cancer/testis antigens and accessory/costimulatory molecules) in solid and hemopoietic human malignancies. However, pharmacologic agents that induce DNA hypomethylation or inhibit histone deacetylation can modify these epigenetic phenomena, restoring the defective expression of selected components of the 'tumor recognition complex' in cancer cells. These antigenic modifications positively modulate the immunogenicity and the immune recognition of cancer cells, making epigenetic drugs attractive agents to design new combined chemoimmunotherapeutic strategies for the treatment of cancer patients. PMID- 14528273 TI - Epigenetic targets in hematopoietic malignancies. AB - Frequent genetic alterations in hematopoietic neoplasias (chromosomal translocations, point mutations, etc.) have provided biologic targets for the development of effective novel therapies. A rapidly increasing body of knowledge provides evidence also for multiple epigenetic alterations in these disorders, which can complement or even precede genetic aberrations. Gene inactivation ('silencing') of tumor suppressor and growth inhibitory genes (e.g. the cyclin dependent kinase inhibitors p16, p15, p21) is frequently mediated by DNA methylation of gene promoters. The acetylation state of histones (functionally linked to the DNA methylation state by the methylcytosine binding protein 2, recruiting histone deacetylases) provides a second major epigenetic silencing mechanism. Therapeutic reversal strategies are being developed for acute leukemias, myelodysplastic syndromes and malignant lymphomas. Since the discovery of the DNA methyltransferase (Dnmt) inhibitory activity of two azanucleosides (5 azacytidine, 5-aza-2'-deoxycytidine/decitabine) even at doses with minimal nonhematologic toxicity, both have been clinically studied in several myeloid neoplasias, particularly in elderly patients unable to tolerate aggressive treatment. Further development of agents counteracting aberrant methylation is directed at more targeted approaches, for example, antisense molecules against Dnmts. Histone deacetylases (HDACs) can be inhibited by numerous compounds (sodium phenylbutyrate, valproic acid, novel compounds such as depsipeptide), which have entered the clinical arena in similar indications as Dnmt inhibitors. Impressive effects of HDAC inhibition in acute promyelocytic leukemia models (PML/RARA expression) translate the finding of HDAC recruitment by this chimeric transcription factor to its target genes. The recent discovery of recruitment by PML/RARA also of Dnmt activity to the retinoic acid receptor-beta promoter makes it an interesting candidate for Dnmt inhibitors. Studies combining a 're expressor' strategy with inhibitors of Dnmts and HDACs are underway. Thus, resensitization to biological agents such as retinoids, colony-stimulating factors and other differentiation inducers may be envisioned. PMID- 14528274 TI - Advantages and limitations of microarray technology in human cancer. AB - Cancer is a highly variable disease with multiple heterogeneous genetic and epigenetic changes. Functional studies are essential to understanding the complexity and polymorphisms of cancer. The final deciphering of the complete human genome, together with the improvement of high throughput technologies, is causing a fundamental transformation in cancer research. Microarray is a new powerful tool for studying the molecular basis of interactions on a scale that is impossible using conventional analysis. This technique makes it possible to examine the expression of thousands of genes simultaneously. This technology promises to lead to improvements in developing rational approaches to therapy as well as to improvements in cancer diagnosis and prognosis, assuring its entry into clinical practice in specialist centers and hospitals within the next few years. Predicting who will develop cancer and how this disease will behave and respond to therapy after diagnosis will be one of the potential benefits of this technology within the next decade. In this review, we highlight some of the recent developments and results in microarray technology in cancer research, discuss potentially problematic areas associated with it, describe the eventual use of microarray technology for clinical applications and comment on future trends and issues. PMID- 14528275 TI - Emerging roles of DNA tumor viruses in cell proliferation: new insights into genomic instability. AB - The small DNA virus proteins E1A and E1B from human Adenovirus, E6 and E7 from human papillomavirus, and large T and small T antigens from SV40, are multifaceted molecular tools that can carry out an impressive number of tasks in the host cell. These viral factors, collectively termed 'oncoproteins' for their ability to induce cancer, can be viewed as paradigmatic oncogenic factors which can disrupt checkpoint controls at multiple levels--they interfere with both 'gatekeeper' cellular functions, including major control pathways of cell cycle and apoptosis, and with 'caretaker' functions, thereby inducing mitotic abnormalities and increasing genomic instability. Both E1A and E7 have been recently found to interact physically with the Ran GTPase. This interaction is key in uncoupling the centrosome cycle from the cell cycle, highlighting a direct link between viral infection and the induction of genomic instability. Further expanding our current knowledge in this field will be crucial to elucidate viral strategies leading to cellular transformation and cancer progression, as well as design novel preventive or therapeutic approaches to human cancer. PMID- 14528276 TI - Viruses and cancer: lessons from the human polyomavirus, JCV. AB - The possible role of eucaryotic viruses in the development of cancer has been the subject of intense investigation during the past 50 years. Thus far, a strong link between some RNA and DNA viruses and various cancers in humans has been established and the transforming activity of several of the viruses in cell culture and their oncogenecity in experimental animals has been well documented. Perhaps, one of the most common themes among the oncogenic viruses rests in the ability of one or more of the viral proteins to deregulate pathways involved in the control of cell proliferation. For example, inactivation of tumor suppressors through their association with viral transforming proteins, and/or impairment of signal transduction pathways upon viral infection and expression of viral proteins are among the key biological events that can either trigger and/or contribute to the process of cancer. In recent years, more attention has been paid to human polyomaviruses, particularly JC virus (JCV), which infects greater than 80% of the human population, due to the ability of this virus to induce a fatal demyelinating disease in the brain, its presence in various tumors of central nervous system (CNS) and non-CNS origin, and the oncogenic potential of this virus in several laboratory animal models. Here, we will focus our attention on JCV and describe several pathways employed by the virus to contribute to and/or accelerate cancer development. PMID- 14528277 TI - Axis of evil: molecular mechanisms of cancer metastasis. AB - Although the genetic basis of tumorigenesis may vary greatly between different cancer types, the cellular and molecular steps required for metastasis are similar for all cancer cells. Not surprisingly, the molecular mechanisms that propel invasive growth and metastasis are also found in embryonic development, and to a less perpetual extent, in adult tissue repair processes. It is increasingly apparent that the stromal microenvironment, in which neoplastic cells develop, profoundly influences many steps of cancer progression, including the ability of tumor cells to metastasize. In carcinomas, the influences of the microenvironment are mediated, in large part, by bidirectional interactions (adhesion, survival, proteolysis, migration, immune escape mechanisms lymph /angiogenesis, and homing on target organs) between epithelial tumor cells and neighboring stromal cells, such as fibroblasts as well as endothelial and immune cells. In this review, we summarize recent advances in understanding the molecular mechanisms that govern this frequently lethal metastatic progression along an axis from primary tumor to regional lymph nodes to distant organ sites. Affected proteins include growth factor signaling molecules, chemokines, cell cell adhesion molecules (cadherins, integrins) as well as extracellular proteases (matrix metalloproteinases). We then discuss promising new therapeutic approaches targeting the microenvironment. We note, however, that there is still too little knowledge of how the many events are coordinated and integrated by the cancer cell, with conspiratorial help by the stromal component of the host. Before drug development can proceed with a legitimate chance of success, significant gaps in basic knowledge need to be filled. PMID- 14528278 TI - Ocular neovascularization: a valuable model system. AB - There is no unique formula for angiogenesis. Instead there is a large group of potential participating proteins that interact in complex ways. Depending upon the surrounding cell types and the relative expression levels of angiogenesis related proteins, the 'angiogenesis cascade' can vary. Therefore, it is valuable to study and compare the role of proteins in several well-characterized vascular beds. The eye provides a useful model system, because it contains several vascular beds sandwiched between avascular tissue. This allows for unequivocal identification and quantitation of new vessels. Retina-specific promoters combined with inducible promoter systems provide a means to regulate the expression of proteins of interest. As a relatively isolated compartment, the eye also provides advantages for gene transfer. By gaining insight regarding the molecular signals involved in various types of ocular angiogenesis, general concepts can emerge that may apply to other settings, including tumor angiogenesis. One concept that has emerged is that despite participation of multiple stimulatory factors for ocular neovascularization, VEGF plays an essential role and interruption of VEGF signaling is an important therapeutic strategy. Another concept is that while most studies have focused on prevention of ocular neovascularization, regression of new vessels is desirable and is achievable with at least three agents, combretastatin A-4 phosphate, pigment epithelium-derived factor, and angiopoietin-2. Finally, endostatin and angiostatin, which have been sources of controversy because of inconsistent results in tumor models, have been shown to have good efficacy when delivered by gene transfer in models of ocular neovascularization. These results provide leads for new ocular treatments and perspective for evaluation of studies of neovascularization in extraocular tissues. PMID- 14528279 TI - Molecular basis of angiogenesis and cancer. AB - Angiogenesis is a term that describes the formation of new capillaries from a pre existing vasculature. This process is very important in physiologic conditions because it helps healing injured tissues, and in female populations it helps forming the placenta after fertilization and reconstructs the inside layer of the uterus after menstruation. Angiogenesis is the result of an intricate balance between proangiogenic and antiangiogenic factors and is now very well recognized as a powerful control point in tumor development. In this particular environment, the fine modulation among proangiogenic and antiangiogenic factors is disrupted, leading to inappropriate vessels growth. In this review, we discuss the molecular basis of angiogenesis during tumor growth and we also illustrate some of the molecules that are involved in this angiogenic switch. PMID- 14528280 TI - Endoglin (CD105): a powerful therapeutic target on tumor-associated angiogenetic blood vessels. AB - Among surface molecules expressed on endothelial cells, endoglin (CD105) is emerging as a prime vascular target for antiangiogenetic cancer therapy. CD105 is a cell membrane glycoprotein mainly expressed on endothelial cells and overexpressed on tumor-associated vascular endothelium, which functions as an accessory component of the transforming growth factor -beta receptor complex and is involved in vascular development and remodelling. Quantification of intratumoral microvessel density by CD105 staining and of circulating soluble CD105 has been suggested to have prognostic significance in selected neoplasias. In addition, the potential usefulness of CD105 in tumor imaging and antiangiogenetic therapy has been well documented utilizing different animal models. PMID- 14528281 TI - Targeted therapy of solid malignancies via HLA class II antigens: a new biotherapeutic approach? AB - Intracellular signals, delivered in professional antigen-presenting cells following the engagement of major histocompatibility complex (MHC) class II molecules, activate a variety of cellular functions that also contribute to efficient antigen presentation. As far as human malignancies, the signaling ability of human leukocyte antigens (HLA) class II molecules is a rather well characterized event in hematologic tumors; in contrast, very limited evidences are available in solid neoplasias of different histotypes that may constitutively express HLA class II antigens. Among solid malignancies, a significant proportion of human cutaneous melanomas have been shown to express HLA class II molecules, and cutaneous melanoma undoubtedly represents a 'model disease' to investigate tumor immunobiology, to unveil the molecular basis underlying the interactions between neoplastic cells and host's immune system, and ultimately to set up new bio-immunotherapeutic approaches. Upcoming preclinical evidences unveil a signaling potential of HLA-DR antigens expressed on melanoma cells, and suggest for the clinical implication of HLA class II molecules as novel therapeutic targets. Therefore, in this review, we will focus on the emerging role of HLA class II antigens as intracellular signal transducing elements in neoplastic cells of the melanocytic lineage, emphasizing their foreseeable role in targeted therapy of human melanoma and potentially of HLA class II antigens-positive tumors of different histology. PMID- 14528282 TI - Biologic and therapeutic role of HER2 in cancer. AB - Overexpression of the human epidermal growth factor-2 (HER2) oncogene in human breast carcinomas has been associated with a more aggressive course of disease. The reason for this association is still unclear, although it has been suggested to rest in increased proliferation, vessel formation, and/or invasiveness. Alternatively, prognosis may not be directly related to the presence of the oncoprotein on the cell membrane, but instead to the breast carcinoma subset identified by HER2 overexpression and characterized by a peculiar gene expression profile. HER2 has also been associated with sensitivity to anthracyclins and resistance to endocrine therapy, suggesting that tyrosine kinase receptor and hormone receptor pathways represent two major proliferation pathways exclusively active in breast carcinomas, one sensitive to chemotherapeutic drugs and the other to antiestrogens. HER2 currently represents one of the most appropriate targets for specific therapy. Indeed, trastuzumab, a monoclonal antibody directed against the extracellular domain of HER2, is therapeutically active in HER2 positive breast carcinomas. However, a consistent number of HER2-positive tumors is not responsive to HER2-driven therapy, indicating the need for a better understanding of the mechanism of action of this new biological drug in vivo. While preclinical studies suggest antibody-dependent cell cytotoxicity as the major mechanism, determination of NK activity at the time of treatment remains mandatory, especially in patients treated with immunosuppressive drugs. The efficacy of prophylactic vaccination has been fully demonstrated in preclinical models, whereas ongoing studies of active immunotherapy using a variety of vaccination regimens against HER2 in tumor-bearing mice and patients have met with only moderate success. PMID- 14528283 TI - The future of antisense therapy: combination with anticancer treatments. AB - The current direction in cancer research is rational drug design, which is based on the evidence that transformed cells are characterized by alterations of genes devoted to the regulation of both cell proliferation and apoptosis. A variety of approaches have been carried out to develop new agents selective for cancer cells. Among these, antisense oligonucleotides (ASOs) are one of such class of new agents able to inhibit specifically the synthesis of a particular cancer associated protein by binding to protein-encoding RNA, thereby preventing RNA function. In the past decade, several ASOs have been developed and tested in preclinical and clinical studies. Many have shown convincing in vitro reduction in target gene expression and promising activity against a wide variety of tumors. However, because of the multigenic alterations of tumors, the use of ASOs as single agents does not seem to be effective in the treatment of malignancies. Antisense therapy that interferes with signaling pathways involved in cell proliferation and apoptosis are particularly promising in combination with conventional anticancer treatment. An overview of the progress of ASOs used in combination therapy is provided. PMID- 14528284 TI - Growth factor receptors as therapeutic targets: strategies to inhibit the insulin like growth factor I receptor. AB - Neoplastic transformation is often related to abnormal activation of growth factor receptors and their signaling pathways. The concept of targeting specific tumorigenic receptors and/or signaling molecules has been validated by the development and successful clinical application of drugs acting against the epidermal growth factor receptor 2 (HER2/neu, Erb2), the epidermal growth factor receptor 1 (EGFR, HER1), the Brc-Abl kinase, the platelet-derived growth factor receptor, and c-kit. This review will focus on the next promising therapeutic target, the insulin-like growth factor I receptor (IGF-IR). IGF-IR has been implicated in a number of neoplastic diseases, including several common carcinomas. From a pharmaceutical standpoint, of particular importance is that IGF-IR appears to be required for many transforming agents (genetic, viral, chemical) to act, but is not obligatory for the function of normal adult cells. The tumorigenic potential of IGF-IR is mediated through its antiapoptotic and transforming signaling, and in some cases through induction of prometastatic pathways. Preclinical studies demonstrated that downregulation of IGF-IR reversed the neoplastic phenotype and sensitized cells to antitumor treatments. The strategies to block IGF-IR function employed anti-IGF-IR antibodies, small molecule inhibitors of the IGF-IR tyrosine kinase, antisense oligodeoxynucleotides and antisense RNA, small inhibitory RNA, triple helix, dominant-negative mutants, and various compounds reducing ligand availability. The experience with these strategies combined with the knowledge gained with current anti-growth factor receptor drugs should streamline the development of anti-IGF-IR therapeutics. PMID- 14528286 TI - Small-molecule cyclin-dependent kinase modulators. AB - Aberrations in cell cycle progression occur in the majority of human malignancies. The main pathway affected is the retinoblastoma (Rb) pathway. The tumor suppressor gene Rb is an important component in the G(1)/S transition and its function is abnormal in most human neoplasms. Loss in Rb function occurs by the hyperactivation of the cyclin-dependent kinases (cdk's). Therefore, modulation of cdk's may have an important use for the therapy and prevention of human neoplasms. Efforts to obtain small-molecule cdk modulators yielded two classes of modulators: direct and indirect modulators. Direct cdk modulators are small molecules that specifically target the ATP binding site of cdk's. Examples for this group include flavopiridol, roscovitine and BMS-387032. In contrast, indirect cdk modulators affect cdk function due to modulation of upstream pathways required for cdk activation. Some examples include perifosine, lovastatin, and UCN-01. The first example of a direct small-molecule cdk modulator tested in the clinic, flavopiridol, is a pan-cdk inhibitor that not only promotes cell cycle arrest but also halts transcriptional elongation, promotes apoptosis, induces differentiation, and has antiangiogenic properties. Clinical trials with this agent were performed with at least three different schedules of administration: 1-, 24- and 72-h infusions. The main toxicities for infusions >/=24-h are secretory diarrhea and proinflammatory syndrome. In addition, patients receiving shorter infusions have nausea/vomiting and neutropenia. A phase II trial of patients with advanced non-small-cell lung carcinoma using the 72-h infusion every 2 weeks was recently completed. The median overall survival for the 20 patients who received treatment was 7.5 months, a survival similar to that obtained in a randomized trial of four chemotherapy regimens containing platinum analogues in combination with taxanes or gemcitabine, or with gefitinib, a recently approved EGFR inhibitor for the treatment of advanced lung cancer. Based on these encouraging results, a phase III trial comparing standard combination chemotherapy versus combination chemotherapy plus flavopiridol is currently under investigation. The second example of direct small-molecule cdk modulator tested in clinical trials is UCN 01 (7-hydroxystaurosporine). UCN-01 has interesting preclinical features: it inhibits Ca(2+)-dependent PKCs, promotes apoptosis, arrests cell cycle progression at G(1)/S, and abrogates checkpoints upon DNA damage. The first phase I trial of UCN-01 demonstrated a very prolonged half-life. Based on this novel feature, UCN-01 is administered as a 72-h continuous infusion every 4 weeks (in second and subsequent cycles UCN-01 is administered as a 36-h infusion). Other shorter schedules (i.e. 3 h) are being tested. Dose-limiting toxicities include nausea/vomiting, hypoxemia, and insulin-resistant hyperglycemia. Combination trials with cisplatin and other DNA-damaging agents are being tested. Recently, phase I trials with two novel small-molecule cdk modulators, BMS 387032 and R Roscovitine (CYC202), have commenced with good tolerability. In summary, novel small-molecule cdk modulators are being tested in the clinic with interesting results. Although these small molecules are directed towards a very prevalent cause of carcinogenesis, we need to test them in advanced clinical trials to determine the future of this class of agents for the prevention and therapy of human malignancies. PMID- 14528285 TI - Notch signaling as a therapeutic target in cancer: a new approach to the development of cell fate modifying agents. AB - Notch signaling controls cell fate decisions including during development and stem cell renewal and differentiation in many postnatal tissues. Increasing evidence suggests that the Notch signaling network is frequently deregulated in human malignancies and that genetic or pharmacological manipulation of Notch signaling is a novel potential strategy for the treatment of human neoplasms. This review article summarizes the most recent preclinical and clinical evidence linking Notch signaling to cancer, delineates questions that remain unanswered and explores potential biopharmacological strategies to manipulate Notch signaling in vivo. PMID- 14528287 TI - Pharmacogenomics: road to anticancer therapeutics nirvana? AB - Interindividual differences in the toxicity and response to anticancer therapies are currently observed for essentially all available treatment regimens. Such 'unpredictable' drug responses are particularly dangerous in the context of anticancer agents that have narrow therapeutic indices. Pharmacogenomics attempts to elucidate the inherited basis of interindividual differences in drug response, with the eventual goal of minimizing such variability through the use of 'individualized' treatments. There are several emerging examples of genetic polymorphisms of drug-metabolizing enzymes, DNA repair genes and drug targets that have been shown to influence the toxicity and efficacy of anticancer treatment. This review discusses the role of genetic variants of UGT1A1, TS and EGFR to exemplify the potential impact of phramacogenomics on the field of anticancer therapeutics. PMID- 14528288 TI - Treatment of non-small-cell lung cancer: state of the art and development of new biologic agents. AB - Conventional treatment of non-small-cell lung cancer (NSCLC) has apparently reached a plateau of effectiveness in improving the survival of NSCLC patients. Although neoadjuvant and adjuvant therapies in early stages are under investigation and some progress has been achieved in the management of locally advanced and advanced disease, treatment outcomes for NSCLC are still to be considered dismal. The majority of patients affected from NSCLC experience metastatic disease and optimization of chemotherapy is unlikely to produce further substantial survival improvement, with symptom relief and quality of life still being the primary goal of treatment. Based on this background, clinical investigation of novel treatment strategies is mandatory. As our understanding of tumor cell biology has increased and several molecular targets for NSCLC have been identified, a number of new biologic agents have been developed. Targeted therapy describes treatment strategies that focus on cell signaling and other biologic pathways involved in tumorigenesis. Several targeted agents have been introduced in clinical trials in NSCLC, the majority in advanced disease, and some phase III studies have already produced definitive results. Currently, the minority of these new agents offer promise of improved outcomes and negative results are more common to be reported than positive ones. However, important lessons can be learned from this first generation of clinical trials that should be considered the first step of clinical research in this field. PMID- 14528289 TI - Pathologic aspects of AIDS malignancies. AB - Since the emergence of the HIV pandemic, a close association between HIV infection and the development of a selected group of cancers has been acknowledged. The introduction of highly active antiretroviral therapy, however, has had a dramatic impact on the incidences of several AIDS-defining malignancies. This suggests the possibility of a direct and indirect role of HIV in HIV-related tumor genesis. The aim of this paper is to review the pathology of AIDS-related malignancies, taking into account the pathogenetic mechanisms and their potential for improving the treatment of these tumors. PMID- 14528290 TI - Therapeutic approaches to AIDS-related malignancies. AB - The introduction of highly active antiretroviral therapy (HAART) has changed dramatically the landscape of HIV disease. Deaths from AIDS-related diseases have been reduced by 75% since protease inhibitor therapy and combination antiretroviral therapy came into use in late 1995. While KS is declining, the situation for non-Hodgkin's lymphoma is more complex with a reduced incidence of primary central nervous system lymphoma, but a relatively stability in the number of patients developing systemic NHL. AIDS related NHL appears not to be markedly decreased by the introduction of HAART and it is the greatest therapeutic challenge in the area of AIDS oncology. The emphasis has now shifted to cure while maintaining vigilance regarding the unique vulnerability of HIV-infected hosts. Furthermore, also for the prolongation of the survival expectancy of these patients, other non AIDS-defining tumors, such as Hodgkin's disease, anal and head and neck, lung and testicular cancer, and melanoma have been recently reported with increased frequency in patients with HIV infection. PMID- 14528291 TI - Visualization of membrane protein domains by cryo-electron microscopy of dengue virus. AB - Improved technology for reconstructing cryo-electron microscopy (cryo-EM) images has now made it possible to determine secondary structural features of membrane proteins in enveloped viruses. The structure of mature dengue virus particles was determined to a resolution of 9.5 A by cryo-EM and image reconstruction techniques, establishing the secondary structural disposition of the 180 envelope (E) and 180 membrane (M) proteins in the lipid envelope. The alpha-helical 'stem' regions of the E molecules, as well as part of the N-terminal section of the M proteins, are buried in the outer leaflet of the viral membrane. The 'anchor' regions of E and the M proteins each form antiparallel E-E and M-M transmembrane alpha-helices, leaving their C termini on the exterior of the viral membrane, consistent with the predicted topology of the unprocessed polyprotein. This is one of only a few determinations of the disposition of transmembrane proteins in situ and shows that the nucleocapsid core and envelope proteins do not have a direct interaction in the mature virus. PMID- 14528292 TI - Covariation of backbone motion throughout a small protein domain. AB - The synchronization (correlation) of conformational fluctuations in folded proteins may influence the rates of enzyme catalysis and ligand binding as well as the stabilities of native proteins and their complexes. However, experimental detection of correlated motions remains difficult. Herein, we present an analysis of the covariation of NMR-derived backbone dynamical parameters among a family of ten mutants of a small protein. Both the spatial restriction and the time scales of backbone motions exhibit a higher degree of covariation than would be expected if the internal motions of each group were independent, providing experimental support for correlated dynamics. Application of this approach to other proteins may reveal dynamical correlations that influence catalysis, ligand-binding and/or protein stability. PMID- 14528293 TI - The structure and binding mode of interleukin-18. AB - Interleukin-18 (IL-18), a cytokine formerly known as interferon-gamma- (IFN-gamma ) inducing factor, has pleiotropic immunoregulatory functions, including augmentation of IFN-gamma production, Fas-mediated cytotoxicity and developmental regulation of T-lymphocyte helper type I. We determined the solution structure of IL-18 as a first step toward understanding its receptor activation mechanism. It folds into a beta-trefoil structure that resembles that of IL-1. Extensive mutagenesis revealed the presence of three sites that are important for receptor activation: two serve as binding sites for IL-18 receptor alpha (IL-18Ralpha), located at positions similar to those of IL-1 for IL-1 receptor type I (IL-1RI), whereas the third site may be involved in IL-18 receptor beta (IL-18Rbeta) binding. The structure and mutagenesis data provide a basis for understanding the IL-18-induced heterodimerization of receptor subunits, which is necessary for receptor activation. PMID- 14528294 TI - Structural and redox plasticity in the heterodimeric periplasmic nitrate reductase. AB - The structure of the respiratory nitrate reductase (NapAB) from Rhodobacter sphaeroides, the periplasmic heterodimeric enzyme responsible for the first step in the denitrification process, has been determined at a resolution of 3.2 A. The di-heme electron transfer small subunit NapB binds to the large subunit with heme II in close proximity to the [4Fe-4S] cluster of NapA. A total of 57 residues at the N- and C-terminal extremities of NapB adopt an extended conformation, embracing the NapA subunit and largely contributing to the total area of 5,900 A(2) buried in the complex. Complex formation was studied further by measuring the variation of the redox potentials of all the cofactors upon binding. The marked effects observed are interpreted in light of the three-dimensional structure and depict a plasticity that contributes to an efficient electron transfer in the complex from the heme I of NapB to the molybdenum catalytic site of NapA. PMID- 14528295 TI - The structure of BtuB with bound colicin E3 R-domain implies a translocon. AB - Cellular import of colicin E3 is initiated by the Escherichia coli outer membrane cobalamin transporter, BtuB. The 135-residue 100-A coiled-coil receptor-binding domain (R135) of colicin E3 forms a 1:1 complex with BtuB whose structure at a resolution of 2.75 A is reported. Binding of R135 to the BtuB extracellular surface (DeltaG(o) = -12 kcal mol(-1)) is mediated by 27 residues of R135 near the coiled-coil apex. Formation of the R135-BtuB complex results in unfolding of R135 N- and C-terminal ends, inferred to be important for unfolding of the colicin T-domain. Small conformational changes occur in the BtuB cork and barrel domains but are insufficient to form a translocation channel. The absence of a channel and the peripheral binding of R135 imply that BtuB serves to bind the colicin, and that the coiled-coil delivers the colicin to a neighboring outer membrane protein for translocation, thus forming a colicin translocon. The translocator was concluded to be OmpF from the occlusion of OmpF channels by colicin E3. PMID- 14528296 TI - The giant electrorheological effect in suspensions of nanoparticles. AB - Electrorheology (ER) denotes the control of a material's flow properties (rheology) through an electric field. We have fabricated electrorheological suspensions of coated nanoparticles that show electrically controllable liquid solid transitions. The solid state can reach a yield strength of 130 kPa, breaking the theoretical upper bound on conventional ER static yield stress that is derived on the general assumption that the dielectric and conductive responses of the component materials are linear. In this giant electrorheological (GER) effect, the static yield stress displays near-linear dependence on the electric field, in contrast to the quadratic variation usually observed. Our GER suspensions show low current density over a wide temperature range of 10-120 degrees C, with a reversible response time of <10 ms. Finite-element simulations, based on the model of saturation surface polarization in the contact regions of neighbouring particles, yield predictions in excellent agreement with experiment. PMID- 14528297 TI - Ex vivo identification, isolation and analysis of tumor-cytolytic T cells. AB - We isolated pure, viable populations of tumor-cytolytic T cells directly from patient blood samples using flow cytometric quantification of the surface mobilization of CD107a-an integral membrane protein in cytolytic granules-as a marker for degranulation after tumor stimulation. We show that tumor-cytolytic T cells are indeed elicited in patients after cancer vaccination, and that tumor reactivity is strongly correlated with efficient T-cell recognition of peptide bearing targets. We combined CD107a mobilization with peptide-major histocompatibility complex (P-MHC) tetramer staining to directly correlate antigen specificity and cytolytic ability on a single-cell level. This showed that tumor-cytolytic T cells with high recognition efficiency represent only a minority of peptide-specific T cells elicited in patients after heteroclitic peptide vaccination. We were also able to expand these cells to high numbers ex vivo while maintaining their cytolytic potential. These techniques will be useful not only for immune monitoring of cancer vaccine trials, but also for adoptive cellular immunotherapy after ex vivo expansion. The ability to rapidly identify and isolate tumor-cytolytic T cells would be very useful in cancer immunotherapy. PMID- 14528298 TI - G13 is an essential mediator of platelet activation in hemostasis and thrombosis. AB - Platelet activation at sites of vascular injury is essential for primary hemostasis, but also underlies arterial thrombosis leading to myocardial infarction or stroke. Platelet activators such as adenosine diphosphate, thrombin or thromboxane A(2) (TXA(2)) activate receptors that are coupled to heterotrimeric G proteins. Activation of platelets through these receptors involves signaling through G(q), G(i) and G(z) (refs. 4-6). However, the role and relative importance of G(12) and G(13), which are activated by various platelet stimuli, are unclear. Here we show that lack of Galpha(13), but not Galpha(12), severely reduced the potency of thrombin, TXA(2) and collagen to induce platelet shape changes and aggregation in vitro. These defects were accompanied by reduced activation of RhoA and inability to form stable platelet thrombi under high shear stress ex vivo. Galpha(13) deficiency in platelets resulted in a severe defect in primary hemostasis and complete protection against arterial thrombosis in vivo. We conclude that G(13)-mediated signaling processes are required for normal hemostasis and thrombosis and may serve as a new target for antiplatelet drugs. PMID- 14528299 TI - Multiple actions of systemic artemin in experimental neuropathy. AB - The clinical management of neuropathic pain is particularly challenging. Current therapies for neuropathic pain modulate nerve impulse propagation or synaptic transmission; these therapies are of limited benefit and have undesirable side effects. Injuries to peripheral nerves result in a host of pathophysiological changes associated with the sustained expression of abnormal pain. Here we show that systemic, intermittent administration of artemin produces dose- and time related reversal of nerve injury-induced pain behavior, together with partial to complete normalization of multiple morphological and neurochemical features of the injury state. These effects of artemin were sustained for at least 28 days. Higher doses of artemin than those completely reversing experimental neuropathic pain did not elicit sensory or motor abnormalities. Our results indicate that the behavioral symptoms of neuropathic pain states can be treated successfully, and that partial to complete reversal of associated morphological and neurochemical changes is achievable with artemin. PMID- 14528300 TI - The antiretroviral enzyme APOBEC3G is degraded by the proteasome in response to HIV-1 Vif. AB - The human protein apolipoprotein B mRNA-editing enzyme-catalytic polypeptide-like 3G (APOBEC3G), also known as CEM-15, mediates a newly described form of innate resistance to retroviral infection by catalyzing the deamination of deoxycytidine to deoxyuridine in viral cDNA replication intermediates. Because DNA deamination takes place after virus entry into target cells, APOBEC3G function is dependent on its association with the viral nucleoprotein complexes that synthesize cDNA and must therefore be incorporated into virions as they assemble in infected cells. Here we show that the HIV-1 virion infectivity factor (Vif) protein protects the virus from APOBEC3G-mediated inactivation by preventing its incorporation into progeny virions, thus allowing the ensuing infection to proceed without DNA deamination. In addition to helping exclude APOBEC3G from nascent virions, Vif also removes APOBEC3G from virus-producing cells by inducing its ubiquitination and subsequent degradation by the proteasome. Our findings indicate that pharmacologic strategies aimed at stabilizing APOBEC3G in HIV-1 infected cells should be explored as potential HIV/AIDS therapeutics. PMID- 14528301 TI - HIV-1 Vif protein binds the editing enzyme APOBEC3G and induces its degradation. AB - The viral infectivity factor (Vif) encoded by HIV-1 neutralizes a potent antiviral pathway that occurs in human T lymphocytes and several leukemic T-cell lines termed nonpermissive, but not in other cells termed permissive. In the absence of Vif, this antiviral pathway efficiently inactivates HIV-1. It was recently reported that APOBEC3G (also known as CEM-15), a cytidine deaminase nucleic acid-editing enzyme, confers this antiviral phenotype on permissive cells. Here we describe evidence that Vif binds APOBEC3G and induces its rapid degradation, thus eliminating it from cells and preventing its incorporation into HIV-1 virions. Studies of Vif mutants imply that it contains two domains, one that binds APOBEC3G and another with a conserved SLQ(Y/F)LA motif that mediates APOBEC3G degradation by a proteasome-dependent pathway. These results provide promising approaches for drug discovery. PMID- 14528302 TI - A domain of Foxn1 required for crosstalk-dependent thymic epithelial cell differentiation. AB - Thymic epithelial cells (TECs) are required for T cell maturation within the thymus. In the nude (Foxn1(nu/nu)) mouse, TECs fail to differentiate. We have generated a hypomorphic allele called Foxn1(Delta), from which an N-terminal domain was deleted. The phenotype was thymus specific, identifying a tissue specific activity for this domain. Foxn1(Delta/Delta) mice showed abnormal thymic architecture, lacking cortical and medullary domains. In contrast to thymi in mice with the null allele, the Foxn1(Delta/Delta) thymus promoted T cell development, but with specific defects at both the double-negative and double positive stages. Thus, initiation and progression of TEC differentiation are genetically separable functions of Foxn1, and the N-terminal domain is required for crosstalk-dependent TEC differentiation. PMID- 14528303 TI - Leukocyte functional antigen 1 lowers T cell activation thresholds and signaling through cytohesin-1 and Jun-activating binding protein 1. AB - Leukocyte functional antigen 1 (LFA-1), with intercellular adhesion molecule ligands, mediates T cell adhesion, but the signaling pathways and functional effects imparted by LFA-1 are unclear. Here, intracellular phosphoprotein staining with 13-dimensional flow cytometry showed that LFA-1 activation induced phosphorylation of the beta(2) integrin chain and release of Jun-activating binding protein 1 (JAB-1), and mediated signaling of kinase Erk1/2 through cytohesin-1. Dominant negatives of both JAB-1 and cytohesin-1 inhibited interleukin 2 production and impaired T helper type 1 differentiation. LFA-1 stimulation lowered the threshold of T cell activation. Thus, LFA-1 signaling contributes to T cell activation and effects T cell differentiation. PMID- 14528304 TI - Enhancement of CIITA transcriptional function by ubiquitin. AB - Although increasing evidence indicates that there is a direct link between ubiquitination and mono-ubiquitination and transcription in yeast, this link has not been demonstrated in higher eukaryotes. Here we show that the major histocompatibility complex (MHC) class II transactivator (CIITA), which is required for expression of genes encoding MHC class II molecules, is ubiquitinated. This ubiquitination enhanced the association of CIITA with both MHC class II transcription factors and the MHC class II promoter, resulting in an increase in transactivation function and in the expression of MHC class II mRNA. The degree of CIITA ubiquitination was controlled by histone acetylases (HATs) and deacetylases (HDACs), indicating that the crucial cellular processes mediated by these enzymes are linked to regulate transcription. Thus, ubiquitin positively regulates a mammalian coactivator by enhancing its assembly at the promoter. PMID- 14528305 TI - Polymorphism for a 1.6-Mb deletion of the human Y chromosome persists through balance between recurrent mutation and haploid selection. AB - Many human Y-chromosomal deletions are thought to severely impair reproductive fitness, which precludes their transmission to the next generation and thus ensures their rarity in the population. Here we report a 1.6-Mb deletion that persists over generations and is sufficiently common to be considered a polymorphism. We hypothesized that this deletion might affect spermatogenesis because it removes almost half of the Y chromosome's AZFc region, a gene-rich segment that is critical for sperm production. An association study established that this deletion, called gr/gr, is a significant risk factor for spermatogenic failure. The gr/gr deletion has far lower penetrance with respect to spermatogenic failure than previously characterized Y-chromosomal deletions; it is often transmitted from father to son. By studying the distribution of gr/gr deleted chromosomes across the branches of the Y chromosome's genealogical tree, we determined that this deletion arose independently at least 14 times in human history. We suggest that the existence of this deletion as a polymorphism reflects a balance between haploid selection, which culls gr/gr-deleted Y chromosomes from the population, and homologous recombination, which continues to generate new gr/gr deletions. PMID- 14528306 TI - The microRNA-producing enzyme Dicer1 is essential for zebrafish development. AB - MicroRNAs (miRNAs) are produced by the Dicer1 enzyme; the role of Dicer1 in vertebrate development is unknown. Here we report target-selected inactivation of the dicer1 gene in zebrafish. We observed an initial build-up of miRNA levels, produced by maternal Dicer1, in homozygous dicer1 mutants, but miRNA accumulation stopped after a few days. This resulted in developmental arrest around day 10. These results indicate that miRNA-producing Dicer1 is essential for vertebrate development. PMID- 14528307 TI - Dicer is essential for mouse development. AB - To address the biological function of RNA interference (RNAi)-related pathways in mammals, we disrupted the gene Dicer1 in mice. Loss of Dicer1 lead to lethality early in development, with Dicer1-null embryos depleted of stem cells. Coupled with our inability to generate viable Dicer1-null embryonic stem (ES) cells, this suggests a role for Dicer, and, by implication, the RNAi machinery, in maintaining the stem cell population during early mouse development. PMID- 14528308 TI - Beta-catenin is critical for dendritic morphogenesis. AB - Regulated growth and arborization of dendritic processes are critical to the formation of functional neuronal networks. Here we identify beta-catenin as a critical mediator of dendritic morphogenesis. We found that increasing the intracellular levels of beta-catenin and other members of the cadherin/catenin complex, namely N-cadherin and alphaN-catenin, enhances dendritic arborization in rat hippocampal neurons, an effect that does not require Wnt/beta-catenin dependent transcription. Conversely, proteins that sequester beta-catenin decreased dendritic branch tip number and total dendritic branch length. Enhancement of dendritic growth elicited by depolarization requires beta-catenin and increased Wnt release. These results identify Wnt/beta-catenin signaling as an important mediator of dendritic development and suggest that the intracellular level of the cadherin/catenin complex is a limiting factor during critical stages of dendritic morphogenesis. PMID- 14528309 TI - Dendritic stability in the adult olfactory bulb. AB - In many regions of the adult mammalian brain, pronounced changes in synaptic input caused by lesions or severe sensory deprivation induce marked sprouting or retraction of neuronal dendrites. In the adult olfactory bulb, adult neurogenesis produces less pronounced, but continuously ongoing synapse turnover. To test the structural stability of adult dendrites in this context, we used two-photon microscopy to image dendrites of mitral and tufted (M/T) cells over prolonged periods in adult mice. Although pharmacologically increased activity could elicit morphological changes, under natural conditions such as ongoing neurogenesis, an odor-enriched environment or olfactory-based learning, M/T cell dendrites remained highly stable. Thus, in a context of ongoing adult synaptogenesis, dendritic stability could serve as a structural scaffold to maintain the organization of local circuits. PMID- 14528310 TI - PI3 kinase enhancer-Homer complex couples mGluRI to PI3 kinase, preventing neuronal apoptosis. AB - Phosphoinositide 3 kinase enhancer (PIKE) is a recently identified nuclear GTPase that activates nuclear phosphoinositide 3-kinase (PI3 kinase). We have identified, cloned and characterized a new form of PIKE, designated PIKE-L, which, unlike the nuclear PIKE-S, localizes to both the cytoplasm and the nucleus. We demonstrate physiologic binding of PIKE-L to Homer, an adaptor protein known to link metabotropic glutamate receptors to multiple intracellular targets including the inositol 1,4,5-trisphosphate receptor (IP3R). We show that activation of group I metabotropic glutamate receptors (mGluRIs) enhances formation of an mGluRI-Homer-PIKE-L complex, leading to activation of PI3 kinase activity and prevention of neuronal apoptosis. Our findings indicate that this complex mediates the well-known ability of agonists of mGluRI to prevent neuronal apoptosis. PMID- 14528311 TI - Production of PtdInsP3 at endomembranes is triggered by receptor endocytosis. AB - Phosphatidylinositol-3,4,5-trisphosphate (PtdInsP(3)) regulates diverse cellular functions, including cell proliferation and apoptosis, and has roles in the progression of diabetes and cancer. However, little is known about its production. Here, we describe fluorescent indicators for PtdInsP(3) that allow a spatio-temporal examination of PtdInsP(3) production in single living cells. After ligand stimulation, PtdInsP(3) levels increased to a larger extent at the endomembranes (that is, the endoplasmic reticulum and the Golgi) than at the plasma membrane. This increase was found to originate from in situ production at the endomembranes, a process stimulated directly by receptor tyrosine kinases endocytosed from the plasma membrane to the endomembranes. The demonstration of PtdInsP(3) production through receptor endocytosis addresses a long-standing question about how signalling pathways downstream of PtdInsP(3) are activated at intracellular compartments remote from the plasma membrane. PMID- 14528312 TI - Targeting of protein ubiquitination by BTB-Cullin 3-Roc1 ubiquitin ligases. AB - The concentrations and functions of many cellular proteins are regulated by the ubiquitin pathway. Cullin family proteins bind with the RING-finger protein Roc1 to recruit the ubiquitin-conjugating enzyme (E2) to the ubiquitin ligase complex (E3). Cul1 and Cul7, but not other cullins, bind to an adaptor protein, Skp1. Cul1 associates with one of many F-box proteins through Skp1 to assemble various SCF-Roc1 E3 ligases that each selectively ubiquitinate one or more specific substrates. Here, we show that Cul3, but not other cullins, binds directly to multiple BTB domains through a conserved amino-terminal domain. In vitro, Cul3 promoted ubiquitination of Caenorhabditis elegans MEI-1, a katanin-like protein whose degradation requires the function of both Cul3 and BTB protein MEL-26. We suggest that in vivo there exists a potentially large number of BCR3 (BTB-Cul3 Roc1) E3 ubiquitin ligases. PMID- 14528313 TI - Enzymatic synthesis of antithrombin III-binding heparan sulfate pentasaccharide. AB - Heparan sulfate (HS) proteoglycans are crucial to numerous biological processes and pathological conditions, but to date only a few HS structures have been synthesized and characterized with regard to structure-function relationships. Because HS proteoglycans are highly diverse in structure, there are substantial limitations on their synthesis by classical chemical means, and thus new methods to rapidly assemble bioactive HS structures are needed. Here we report the biosynthesis of bioactive HS oligosaccharides using an engineered set of cloned enzymes that mimics the Golgi apparatus in vitro. We rapidly and efficiently assembled the antithrombin III-binding pentasaccharide in just 6 steps, in contrast to the approximately 60 steps needed for its chemical synthesis, with an overall yield at least twofold greater and a completion time at least 100 times faster than for the chemical process. PMID- 14528314 TI - The genome sequence of the entomopathogenic bacterium Photorhabdus luminescens. AB - Photorhabdus luminescens is a symbiont of nematodes and a broad-spectrum insect pathogen. The complete genome sequence of strain TT01 is 5,688,987 base pairs (bp) long and contains 4,839 predicted protein-coding genes. Strikingly, it encodes a large number of adhesins, toxins, hemolysins, proteases and lipases, and contains a wide array of antibiotic synthesizing genes. These proteins are likely to play a role in the elimination of competitors, host colonization, invasion and bioconversion of the insect cadaver, making P. luminescens a promising model for the study of symbiosis and host-pathogen interactions. Comparison with the genomes of related bacteria reveals the acquisition of virulence factors by extensive horizontal transfer and provides clues about the evolution of an insect pathogen. Moreover, newly identified insecticidal proteins may be effective alternatives for the control of insect pests. PMID- 14528315 TI - Targeting cytokines to inflammation sites. AB - To increase the half-life of a cytokine and target its activation specifically to disease sites, we have engineered a latent cytokine using the latency-associated protein (LAP) of transforming growth factor-beta 1 (TGF-beta 1) fused via a matrix metalloproteinase (MMP) cleavage site to interferon (IFN)-beta at either its N or C terminus. The configuration LAP-MMP-IFN-beta resembles native TGF-beta and lacks biological activity until cleaved by MMPs, whereas the configuration IFN-beta-MMP-LAP is active. LAP provides for a disulfide-linked shell hindering interaction of the cytokine with its cellular receptors, conferring a very long half-life of 55 h in vivo. Mutations of the disulfide bonds in LAP abolish this latency. Samples of cerebrospinal fluid (CSF) or synovial fluid from patients with inflammatory diseases specifically activate the latent cytokine, whereas serum samples do not. Intramuscular injection in arthritic mice of plasmid DNA encoding these constructs demonstrated a greater therapeutic effect of the latent as compared to the active forms. PMID- 14528317 TI - Optimized lentiviral vector production and purification procedure prevents immune response after transduction of mouse brain. AB - HIV-derived lentiviral vectors are efficient vehicula to deliver genes into the brain and hold great promise for future gene therapy of neurodegenerative disorders. However, administration of the current vector preparations in mouse brain was found to induce a systemic immune response to vector proteins and a modest inflammation in the brain. Moreover, serum antibodies from vector-treated animals were capable of partially neutralizing lentiviral vector-mediated transduction in cell culture. To avoid this unexpected immune reaction, we have optimized new vector production and purification protocols. Purification by sucrose gradient ultracentrifugation abolished the immune response, but vector titers also decreased substantially. Lentiviral vector production in the absence of serum in the cell culture medium equally reduced immunogenicity without affecting transduction efficiency. These results have important implications for future clinical use of lentiviral vectors, and for the use of lentiviral vectors to create animal models for neurodegenerative diseases that have an important neuroinflammatory component. PMID- 14528318 TI - Human adenovirus type 35: nucleotide sequence and vector development. AB - In this report, we describe the complete 34,794 base pair genomic sequence of the human adenovirus serotype 35 (Ad35) Holden strain. The viral genome exhibits a compact organization similar to other adenoviral serotypes, with overlapping genes on both strands. In all, 47 open reading frames (ORFs) were identified, including early (E1, 2, 3, 4) and late (L1, 2, 3, 4, 5) regions conserved among the adenoviridae family. In addition, 14 ORFs were identified that do not encode known adenoviral genes. Comparison of the predicted translational products of the conserved genes with those of other adenoviruses revealed that Ad35 has high homology to Ad7, Ad3, Ad21, Ad17, and simian Ads25. Based on the complete Ad35 DNA sequence, E3-, E1-, and E1/E3-deleted Ad35-based vector systems were developed. An HEK293-derived cell line was established for the propagation of the E1-deleted Ad35 vector, avoiding the emergence of replication-competent adenovirus. Moreover, production of the E1-deleted recombinant Ad35 vector was achieved by transient transduction of a plasmid encoding the Ad35 E1B gene in HEK293 cells. Testing showed that the Ad35-based vector efficiently infects both human and rhesus macaque dendritic cells. Our novel Ad35-based vectors and their corresponding packaging cell lines will provide a versatile and powerful system for DNA-based vaccine development and gene therapy applications. PMID- 14528319 TI - Adenoviral delivery of soluble VEGF receptor 1 (sFlt-1) abrogates disease activity in murine collagen-induced arthritis. AB - The angiogenic factor VEGF promotes synovitis and bone erosion in rheumatoid arthritis (RA). Previously, we have demonstrated that VEGF expression correlates with disease severity in RA patients and in murine collagen-induced arthritis (CIA). In this study, we adopted an adenoviral gene delivery system expressing soluble VEGF receptor 1 (sFlt-1) to further study the role of VEGF in CIA. Arthritis was induced in DBA/1 mice by injection of bovine collagen. Adenoviruses expressing human soluble VEGF receptor 1 (AdvsFlt-1), or without transgene (Adv0), were injected intravenously on the first day of arthritis. We found that disease severity and paw swelling were significantly suppressed in mice receiving AdvsFlt-1, when compared to untreated or Adv0-treated mice. Expression of sFlt-1 peaked 24 h after injection, with protein detectable in the liver, synovial issue and serum, but rapidly decreased by 72 h. The effect of sFlt-1 expression on signs of disease was paralleled by reduced joint destruction and decreased expression of the vascular marker von Willebrand factor. In summary, adenoviral delivery of human soluble VEGF receptor type 1 significantly suppressed disease activity in CIA. The actions of AdvsFlt-1 are likely to be mediated by reduced synovial neovascularization, and these results support the concept that VEGF blockade may be an effective therapeutic adjunct for the treatment of RA. PMID- 14528320 TI - Adenovirus-mediated transfer of the PTEN gene inhibits human colorectal cancer growth in vitro and in vivo. AB - The tumor-suppressor gene PTEN encodes a multifunctional phosphatase that is mutated in a variety of human cancers. PTEN inhibits the phosphatidylinositol 3 kinase pathway and downstream functions, including activation of Akt/protein kinase B (PKB), cell survival, and cell proliferation in tumor cells carrying mutant- or deletion-type PTEN. In such tumor cells, enforced expression of PTEN decreases cell proliferation through cell-cycle arrest at G1 phase accompanied, in some cases, by induction of apoptosis. More recently, the tumor-suppressive effect of PTEN has been reported in ovarian and thyroid tumors that are wild type for PTEN. In the present study, we examined the tumor-suppressive effect of PTEN in human colorectal cancer cells that are wild type for PTEN. Adenoviral-mediated transfer of PTEN (Ad-PTEN) suppressed cell growth and induced apoptosis significantly in colorectal cancer cells (DLD-1, HT29, and SW480) carrying wtPTEN than in normal colon fibroblast cells (CCD-18Co) carrying wtPTEN. This suppression was induced through downregulation of the Akt/PKB pathway, dephosphorylation of focal adhesion kinase (FAK) and mitogen-activated protein kinase (MAPK) and cell-cycle arrest at the G2/M phase, but not the G1 phase. Furthermore, treatment of human colorectal tumor xenografts (HT-29, and SW480) with Ad-PTEN resulted in significant (P=0.01) suppression of tumor growth. These results indicate that Ad-PTEN exerts its tumor-suppressive effect on colorectal cancer cells through inhibition of cell-cycle progression and induction of cell death. Thus Ad-PTEN may be a potential therapeutic for treatment of colorectal cancers. PMID- 14528321 TI - Ocular transfer of retinal glial cells transduced ex vivo with adenovirus expressing viral IL-10 or CTLA4-Ig inhibits experimental autoimmune uveoretinitis. AB - Gene transfer using immunomodulatory molecules is a promising tool for in vivo regulation of immune responses. Experimental autoimmune uveitis (EAU), which serves as a model for human ocular inflammation, is induced by systemic immunization with autoantigens, but its expression is restricted to the eye. Previously, we reported protection of rodents against EAU by intravenous or/and periocular injection of vIL-10-expressing adenovirus. Here, the expression of vIL 10 was targeted into the rat Lewis eye, by intravitreal injection of either the free virus or ex vivo transfected retinal Muller glial cells (RMG-vIL-10). As shown using GFP-expressing adenovirus, a longer expression of transgene was observed in the eye after transfer of transfected syngeneic RMG cells than was seen after injection of free virus. Intravitreal injection of RMG-vIL-10 led to significant decrease in ocular pathological manifestations, compared to control RMG cells. This was observed when cells were injected simultaneously with autoantigen, but also after a delayed administration of transfected cells. Finally, injection of RMG cells transfected with adenovirus expressing CTLA4 had a strongly protective effect. In conclusion, inhibition of antigen presentation at the site of expression of the autoimmune disorders represents an attractive alternative to treat ocular inflammation, and the transfer of ex vivo genetically modified cells provides a promising method to target the factor of interest into the eye. PMID- 14528322 TI - Conditionally replicative adenovirus expressing a targeting adapter molecule exhibits enhanced oncolytic potency on CAR-deficient tumors. AB - Conditionally replicative adenoviruses (CRAds) are potentially useful agents for anticancer virotherapy approaches. However, lack of coxsackievirus and adenovirus receptor (CAR) expression on many primary tumor cells limits the oncolytic potency of CRAds. This makes the concept of targeting, that is, redirecting infection via CAR-independent entry pathways, relevant for CRAd development. Bispecific adapter molecules constitute highly versatile means for adenovirus targeting. Here, we constructed a CRAd with the Delta24 E1A mutation that produces a bispecific single-chain antibody directed towards the adenovirus fiber knob and the epidermal growth factor receptor (EGFR). This EGFR-targeted CRAd exhibited increased infection efficiency and oncolytic replication on CAR deficient cancer cells and augmented lateral spread in CAR-deficient 3-D tumor spheroids in vitro. When compared to its parent control with native tropism, the new CRAd exhibited similar cytotoxicity on CAR-positive cancer cells, but up to 1000-fold enhanced oncolytic potency on CAR-deficient, EGFR-positive cancer cells. In addition, EGFR-targeted CRAd killed primary human CAR-deficient brain tumor specimens that were refractory to the parent control virus. We conclude, therefore, that CRAds expressing bispecific targeting adapter molecules are promising agents for cancer treatment. Their use is likely to result in enhanced oncolytic replication in cancerous tissues and thus in more effective tumor regression. PMID- 14528324 TI - The World Summit on Social Development, health, and Latin America. PMID- 14528323 TI - Downregulation of TNF-alpha and VEGF expression by Sp1 decoy oligodeoxynucleotides in mouse melanoma tumor. AB - Melanoma tumor growth and progression are highly dependent on adequate blood supply through angiogenesis. Since several genes involved in angiogenesis revealed potential binding sites for the transcription factor Sp1, we have examined the effects of local inoculation of Sp1 decoy oligodeoxynucleotides (ODNs) on the growth of transplanted murine melanoma tumors and the expression of VEGF and TNF-alpha within these tumors. Treatment with Sp1 decoy ODNs, but not their mutated form, led to a significant increase (P=0.041) of the tumor necrotic area, as evaluated morphometrically. Tumor necrosis was associated with a significant decrease of microvascular density (P=0.012) and relative vascular area (P=0.026), as determined by counting CD34-positive vascular structures within the tumor microenvironment of Sp1 decoy ODNs and control ODN-treated tumors. RT-PCR experiments showed a strong decrease in the levels of VEGF188 and VEGF164 isoforms and a moderate decrease of TNF-alpha in Sp1 decoy-treated tumors. Taken together, our results indicate that Sp1 decoy ODNs may inhibit angiogenesis by affecting the gene expression of key players in angiogenesis such as TNF-alpha and VEGF. These findings indicate that Sp1 decoy ODNs may be a potential new therapeutic tool in antiangiogenic therapy. PMID- 14528325 TI - [The inadequacy of threshold values for preventing lead poisoning in Brazil]. AB - This article reviews studies performed since 1975 on the occurrence of central and peripheral neurological manifestations in low-level lead exposure. The review shows that in many workers exposed to lead who present indications of both biological effects and exposure below the limits established by the Brazilian laws, abnormalities are found in peripheral nerve conduction velocity and also in several central nervous system functions. The study thus suggests that the threshold values used in Brazil to confirm lead poisoning should be revised. PMID- 14528326 TI - [Arterial hypertension in Ilha do Governador, Rio de Janeiro, Brazil: I. Methodology]. AB - The aim of this paper (first of a series of two) is to present the main methodological aspects used in a cross-sectional study whose proposals were to establish the prevalence of arterial hypertension in adults in Ilha do Governador (a district of the city of Rio de Janeiro, Brazil), and to examine its relationship to other risk factors, including those for cardiovascular diseases, so as to establish its control status. Sampling design (of clusters in strata, according to mean household income) is described in detail, in addition to measurement methods, and training of observers. Indicators of performance in field work and quality control are also presented as results of the application of the metodology of study. PMID- 14528327 TI - [Health policy and legislation concerning traditional indigenous medicine in Mexico]. AB - Various Mexican federal and state government agencies dealing with indigenous affairs are trying to establish control over traditional medicine and by recognizing it achieve cost decreases for health care directed towards Indian groups, bolstering it as a first-care level. The result is that the community absorbs the costs of medical care, leading to a reduction in the number of patients seeking government-run health care services. Traditional indigenous healers and their organizations consider the law a two-edged sword. On the one hand it provides them with legal measures for their protection, but on the other, they perceive it as a dangerous threat to their culture and practices. This has led them to seek legal advice, training, and active participation in the drafting process for such legislation. The result of this project is that they have been able to learn about the law, discuss their demands, and work out a proposal which is included in this document. PMID- 14528328 TI - [Diagnosis of health conditions in a pan-mining community in the Tapajos River Basin, Itaituba, Par , Brazil, 1992]. AB - The purpose of this article is to describe health conditions in a pan-mining community, in particular the interaction of mercury contamination with other health problems. This study was carried out in a panning mine located on Rato Creek, a tributary of the Tapajo s River Basin in the municipality of Itaituba, State of Par The study population consisted of 223 individuals. An epidemiological questionnaire was applied, medical treatment was given, and biological specimens were collected. Testing of fecal samples showed that 96.1% of the individuals had parasites and 66.4% were anemic. Prevalence of hepatitis B virus was 85.0%, and 11 individuals (6.0%) were virus carriers (HBsAg positive). Of 186 patients tested for malaria, 65 individuals (35.0%) had the infection, of whom 34 (52.3%) were asymptomatic. Prevalence of syphillis was 41.6%. Urine mercury levels were measured in 173 individuals. Some 16 (9.2%) of those tested had mercury levels between 10 and 19 ug/l, while 9 patients (5.2%) had levels above 20 ug/l. An overview of general health conditions showed a combination of problems in individuals, who lacked adequate diagnosis, in addition to a high level of self-medication, given the absence of health facilities. In addition, these individuals are threatened by critical levels of mercury contamination. In conclusion, this article gives a general overview of health conditions in these pan-mining communities, demonstrating the multicausal nature of health conditions in the Amazon region, a fact which should be considered when planning appropriate strategies for clinical treatment. PMID- 14528329 TI - [Brazilian physician Nina Rodrigues: analysis of a scientific career]. AB - One of the most productive ways of investigating the history of Brazilian medical practice at the turn of this century is to focus on the fruitful and ambiguous careers of those physicians that played an important role in the changes affecting the medical field at that time. This paper aims at analyzing the scientific career of physician-anthropologist Raimundo Nina Rodrigues, taking as its conceptual basis Pierrre Bourdieu's notion of "scientific field ". The paper's point of departure is the notion that the medical field is a space of confrontation that is structurally determined by previous conflicts in which physicians try to monopolize claims to scientific authority and competence. Nina Rodrigues' medical profile appears in this context as a precise indicator of the process of specialization and competition that took place during that period. PMID- 14528330 TI - [Linear growth retardation and classroom performance of children from the city of Osasco, Sao Paulo, Brazil]. AB - The objective of the present study was to investigate the significance of linear childhood growth retardation in relation to classroom performance. It began with a Height Census carried out in the 1989 school year, involving children attending the first grade of all public and private schools in Osasco (Greater S o Paulo Metropolitan Area, Brazil), which identified the presence of growth retardation. Using a prospective study, classroom performance was evaluated throughout the school year in 170 children entering school and characterized by the height-for age indice below -2 z scores (NCHS/OMS reference population) and in 205 children entering school and characterized by height-for-age above -1 z score. Classroom performance of stunted school children was lower than that of students without growth retardation. The study indicated that increased risk of school failure of those students remained the same, even after adjustment for possible confounding variables (present nutritional status and socioeconomic variables). PMID- 14528331 TI - [Anthropometric profile of children aged 0 to 5 years in the city of Niteroi, Rio de Janeiro, Brazil]. AB - The Niteroi-based Nutritional and Food Surveillance System (SISVAN Niteroi, RJ, Brazil) performed an anthropometric survey which included 2,248 children from zero to 60 months of age in September 1992, during a mass vaccination day program. The purpose was to determine the nutritional status of children from that city. The indexes used were Weight for Age (WA), Weight for Height (WH), and Height for Age (HA), and the reference population was that of the NCHS (National Center for Health Statistics, USA), adopted and recommended by the WHO. The indexes were analyzed using the classifications by Gomez and Z-Scores. Prevalence of undernutrition was calculated on the basis of the proportion of children with Z-scores under -2.00, using HAZ to stabilish chronic forms and WHZ for acute ones. A data base was built using Epi Info, version 5.0 1b, and a special program from CDC (Centers for Disease Control) was used to calculate anthropometric indexes. The results showed as much as 20.1 % of undernutrition, including all forms from the Gomez classification. In the city as a whole, 6.8% had HAZ under 2.00. The results indicated a serious situation of chronic undernutrition that demands continuous nutritional surveillance in Niteroi, and especially in the city's Eastern District. PMID- 14528332 TI - [Environmental and social determinants in schistosomiasis mansoni in Ravena, Minas Gerais, Brazil]. AB - This study identified the role of biological and social determinants in the transmission of schistosomiasis mansoni in Ravena, Minas Gerais, Brazil, in 1980. This data was used to characterize the clinical and epidemiological profiles of the endemic desease in the population, allowing for the determination of the efficacy of the potable water supply and the specific treatment of those infected with S. mansoni. The district contains three locations, Ravenopolis, Ravena and Lavapes, where the prevalence of the endemic disease was, 20.1%, 42.6% and 63.9%, respectively. The prevalence in the district was statistically higher in men. The age brackets that displayed differences by gender were 10-14 and 15-19 years. Severity of infection was statistically different among individuals within the 10 14 year bracket in ali three locations, and in the 15-19 year bracket among individuals from Ravenopolis and Ravena. The hepatointestinal form was associated with age, and individuals under 15 years of age presented risk of infection 8.85 times higher than adults. Multivariable analysis of the factors involved in transmission of the disease showed that Lavapes was independently associated with infection. In that area, poor sanitary conditions and the proximity of houses to streams infested with S. marsoni cercariae facilitated infection of neighborhood women while performing domestic activities, as well as men digging sand from the streams for construction. These results show the focal nature of transmission of the endemic requiring specific intervention for effective control of disease. PMID- 14528333 TI - [Triatoma brasiliensis Neiva, 1911 (Hemiptera, Reduviidae): observations on dynamics of feeding behavior in an artificial ecotope]. AB - In order to study the behavior of triatomine bugs under artificial ecotopic conditions, an experimental, double-walled poultry coop was built. Its inside walls were made of sun-dried clay blocks which could be broken apart for better access to the insects hiding in their crevices. The coop was subdivided into two main areas of different sizes using wire netting and a wall made of perforated bricks. The chickens, the only food source available for the triatomines, were kept in the larger, rectangular area, further subdivided diagonally. The triatomines were released in the smaller area and were allowed to migrate to the larger one though the holes in the bricks. Information about migration, survival, and hiding places of fourth-stage nymphs of Triatoma brasiliensis was thus gathered. At 30-day intervals, over a period of six months, the inner walls of the house were taken apart and inspected. The nymphs were marked with different colors according to the month and place of capture. It was found that 88.5% of the nymphs were captured near the roost and found engorged. After having taken their first blood meal, they would remain in the vicinity of their food source. The results obtained are considered epidemiologically relevant, and particularly so as pertaining to the control of the vectors of Chagas disease. PMID- 14528334 TI - [Risk factors for unfavorable birth weight in areas surrounding Guadalajara, Mexico]. AB - This study seeks to identify the incidence of unfavorable birth weight (UBW) -< 3000 grams, the factors associated with this condition, and the probability that a child has an UBW in the presence or absence of identified risk factors, in Tlaquepaque and Tonal , outlying areas of Greater Metropolitan Guadalajara, Mexico. A sample of live-born infants in 1991, children of mothers covered by the Mexican Institute of Social Security were selected from the study area; through multistage probabilistic sampling, random selection was made of Family Medical Units within the study areas, and of physicians' offices within those selected units; finally, all the liveborn infants in 1991 from these selected physicians' offices were studied: a total of 141 newborns were studied Mothers of the newborns chosen were interviewed; a questionnaire with different biomedical, socioeconomic, and demographic items was applied by social workers specially trained for the purpose. Logistic regression models were used lo estimate odds ratios (OR), with 95% confidence intervals (CI). The probability that a child would have UBW in the presence or absence of identified risk factors were also calculated. The results show that 22% of the infants studied were born weighing less than 3000 grams; four risk factors were statistically associated with UBW: mother's age of 35 or older (OR=18.47, CL 1.86-83.54); mother worked outside the home (OR= 3.14, C1:1.15-8.59); mother's pre-pregnancy low weight (OR= 5.04, CL1.04-24.47); and late detection of pregnancy (OR=2.64, CI: 1.02-6.84). In the presence of all the risk factors identified, there is a very high probability (0.97) that a child be born with birthweight less than 3000 g, and in the absence of these factors the probability is reduced substantially (0.04). The findings indicate the magnitude of the problem studied, but also the possibility of health services acting in a timely fashion, since the identified risk factors make it possible lo predict, with relative certainty, the birth of a child weighing less than 3000 g. PMID- 14528335 TI - [Women, health, and labor in Brazil: challenges for new action]. AB - Despite the remarkable rise in women's participation in the labor market in Brazil, its consequences on health are still virtually unknown. This study aims to identify theoretical and methodological problems in the relationship between labor and women's health from a gender perspective. Characteristics of women's occupational placement are described and analyzed as resulting from their role in social reproduction. The study examines the development of several conciliatory strategies between paid work and housework which are discussed as potential determinants of health problems and support the need for a critical reappraisal of theoretical and methodological strategies to reach a better understanding of the complexity and specificities of women's living and working conditions. The author also stresses the role of women's recent participation in the trade union movements in defense of health, body rights, and women's issues in the workplace, as well as the need for a new framework embodied in the women's social movement. The study thus points to the challenge to produce knowledge on this subject in order to unveil the uniqueness of the national scenario marked by unemployment, informal jobs, low salaries, weak trade unions and other civil organizations, and traditional domestic and marriage relationships. PMID- 14528336 TI - [Comments on models for studying human rabies outbreaks from bats]. AB - This paper analyzes which of the possible public health models could contribute to an understanding of human rabies outbreaks transmitted by bats. First, we present a short review of the most common pertinent models. Second, we present an investigation of a human rabies outbreak transmitted by bats based on the socio epidemiological model. The same outbreak is analyzed again using the complex system theory as a theoretical framework, as both a theoretical-practical exercise seeking an approximation to this theory and a model to study this problem. PMID- 14528337 TI - [Studies on women in three public health journals: a bibliographical review]. AB - The purpose of this study is determine how the First National Conference on Women's Health Rights, held in Brasilia in 1986, was reflected in three main Brazilian public health journals. The articles were published in Revista de Saude Publica (Faculdade de Saude Publica, Universidade de Sao Paulo), Saude em Debate (Centro Brasileiro de Estudos de Saude) and Cadernos de Saude Publica (Escola Nacional de Saude Publica, Fundacao Oswaldo Cruz), from 1986 to 1993. PMID- 14528338 TI - [Solid waste and nosocomial waste: an ethical discussion]. AB - This paper deals with issues pertaining to solid waste. It discusses problems resulting from both the life style of modern civilization and relationships between the First and Third Worlds, as well as prospects for the environment if a new ethic of solidarity among human beings and respect for life is not established Beginning with this overview, the article deals with specific issues of hospital waste and the interference and impositions of First-World "culture" in the systems for handling, processing, and disposing of hospital waste in Brazil. The article concludes by analyzing whether or not such nosocomial waste should be considered hazardous. PMID- 14528339 TI - [Visceral leishmaniasis in the State of Maranhao, Brazil: evolution of an epidemic]. AB - The authors provide a brief report on the historical evolution of visceral leishmaniasis in the State of Maranhao, Brazil, evaluating possible factors for growth of the disease in the State and control measures by the Brazilian Ministry of Health to integrate health services into the maintenance of control programs. PMID- 14528340 TI - [Hepatosplenic forms of schistosomiasis mansoni in the interior of Northeastern Brazil]. AB - Field studies on the morbidity of schistosomiasis mansoni were carried out in some rural endemic areas of Northeastern Brazil during the first semester of 1991. These cross-sectional studies were limited to the assessment of the gradient of clinical forms of the disease. The entire population present in a house-to-house survey was examined. Results have shown a reduction in the prevalence rates of the more severe form of schistosomiasis, the hepatosplenic form, when compared with previous similar studies made in the same areas during the 1960s and 1970s. Changes in prevalence rates of the severe clinical forms from the younger to the older age groups were observed. PMID- 14528341 TI - On the controversy about the efficacy of the antimeningococcal B vaccine: methodological pitfalls. AB - The present communication analyses methodological problems related to evaluating the protection given by the anti-meningococcal B vaccine to children under four years of age in a case-control study carried out in Sao Paulo, Brazil. Detection of a hospital selection bias, among other disturbing features of the study, suggests that the efficacy of the vaccine was underestimated. Altogether, data presented support a wider use of the Cuban-produced vaccine to reduce morbidity and mortality from serogroup B meningococcal disease. PMID- 14528346 TI - [Not Available]. PMID- 14528347 TI - [Rethinking the challenges of Ulysses and Faust: health, the individual and history]. AB - Concentrating on two historical synecdoches, Ulysses and Faust, this article takes up the etymology of the Latin term salute as a unit lying somewhere between "existential needs" and "especially human needs", leading to the challenge of satisfying the need for "conservation of life" and at the same time to "surpass it, go beyond it". Both meanings are present in Ulysses attitude of not succumbing to the siren s melody and in Faust s desire to rise above everyday life. Some aspects of the Marxist conception of the philosophy of history and Althusser s Structuralist Marxism are criticized. Also, in light of Marxist Hellerian theory, the article analyzes the hypothesis that the target of the final reports of the VIII National Health Conference was "particular man" and not the individual, since health is treated only as "an existential need", and does not envisage the generic human. As a theoretical challenge, the text, in search of the unfolding individual, finally recommends the construction of "epistemological sutures" between nature and society, everyday life and universality, and the young and old Marx. PMID- 14528348 TI - [Debate on the paper by Melo-Filho]. PMID- 14528349 TI - [Debate on the paper by Melo-Filho]. PMID- 14528350 TI - [Debate on the paper by Melo-Filho]. PMID- 14528352 TI - [Debate on the paper by Melo-Filho]. PMID- 14528351 TI - [Debate on the paper by Melo-Filho]. PMID- 14528353 TI - [Debate on the paper by Melo-Filho]. PMID- 14528355 TI - [Social and political actors in the formulation of health reform in Spain]. AB - An analysis of the participation of social and political actors in the process of formulating health policy allows one to understand the specific characteristics of the organization and operation of a health system. This study analyzes the drafting process for the General Health Act (LGS) in Spain with the purpose of establishing the relationship between social, political, and economic actors in both the formulation of the Act itself and the organization of the Spanish Health System. A case study was carried out from 1982 through 1986. Documentary parliamentary data, the medical press, national magazines and journals, and press reports by political, social, and public health actors were analyzed. The first version of the General Health Act presented by the Spanish Socialist Workers' Party (PSOE) proposed a health system with funding and public administration aimed at achieving universal health coverage, integrated care, community participation, and health education. This proposal was submitted to a complex negotiating process with business groups, unions, and health professionals. The General Health Act as finally approved excludes the principles of equity and incorporates private interests in health: "free choice of doctor and hospital", public funding and private administration of the health system, and the establishment of Social Security as the core of the entire health system. PMID- 14528356 TI - [Urban schistosomiasis and social and epidemiological heterogeneity in Campinas, Sao Paulo, Brazil]. AB - This study deals with the relationship between schistosomiasis and urbanization, particularly the urbanization process experienced by cities in dependent capitalist countries, with large groups of people in precarious living conditions. Six hundred and ten notified autochthonous cases in Campinas during the period from 1970 to 1990 are analyzed through notification files with regard to patients socio-economic conditions. Autochthonous cases of schistosomiasis were demonstrated as having a more heterogeneous geographical distribution within the urban area than could be inferred from previous studies that correlate the occurrence of transmission with precarious sanitary conditions. PMID- 14528357 TI - [Work in medicine: some issues concerning professional autonomy]. AB - The author analyzes several issues pertaining to medical practice as related to the technical autonomy characterizing professional activity in this field. She points to the coexistente of a scientific side and an empirical/pragmatic side of medicine, the latter known as the medical art. The article also discusses the complexity of medical command of technological rationality which presides over the practical use of scientific knowledge. The author defines work in medicine as a "tensive structure" and examines its consequences for self-representation by physicians, their daily work, and themselves as agents of professional labor and subjects of action. PMID- 14528358 TI - [The role of stress in exhaustion among female workers]. AB - The purpose of this paper has the purpose to compare profiles for exhaustion between different groups of working women, showing the importance of stress as the result of gender conditions. In the research projects reviewed dealing with work processes and related diseases, a close relation was, found between stress and health problems. Some of them demonstrate that domestic and job demands lead to continuous effort and insufficient replacement of working capacity. In addition, since women cannot meet both demanding tasks, they develop persistent anxiety, and the most immediate repercussions are fatigue and stress-related disease. PMID- 14528359 TI - [District allocation and utilization of health care services in Pau da Lima, Salvador, Bahia, Brazil]. AB - This cross-sectional study is aimed at the identification of patterns in the utilization of health care services in Pau da Lima, a neighborhood in the city of Salvador, Bahia. In 1992, a household survey was carried out with 384 families selected through a random cluster sampling design. In each household, the family head or a surrogate informant was asked to answer a questionnaire about their use of any type of health care in the last month. From the total of 1,887 individuals, 236 reported at least one visit to health facilities during the referent period This means an estimated prevalence of health services utilization of 12.5%. With regard to the type of services, 25.9% of all visits were made to public sector facilities. Attendants were mainly women (73.7%) from 15 to 29 years of age. The main reason reported for the choice of the health facility was availability (63.7%), and the major reason for seeking health care was disease related conditions (75.7%), rather than preventive procedures. These findings may result from the still unfinished process of distritalizacao(district allocation of services), which is evident in the lack of health services' infrastructure or coverage, as well as in persistent organizational problems. These issues may lead the population living in the respective district to seek health care outside the assigned catchment area. The authors discuss these findings in the context of the expansion of the private sector in the health area and the financial collapse of the public budget in Brazil. PMID- 14528360 TI - [Prevalence of hypovitaminosis a in children of peripheral districts of Campinas Sao Paulo, Brazil]. AB - The prevalence of hypovitaminosis A among children of the peripheral districts of the city of Campinas, S o Paulo, Brazil, was estimated by determining serum retinol levels by High Performance Liquid Chromatography (HPLC) in a sample of 131 children aged between three and ten years, between April 1991 and February 1992. A prevalence of 17.6% and retinol concentrations in the range of 0.35 to 0.70 micromol/L were found (CI=11.1-24.1; 95%), indicating the existence of public health risk Ophthalmological examinations, however, failed to detect any cases of xerophthalmy. Additional characterization of the sample was obtained from 341 children. The per capita income of the average household was surprisingly high for low-income areas. According to FAO-WHO standards, food consumption was adequate only for protein (133.96%). Adequacy levels were low for energy (87.76%) and particularly for vitamin A (66.13%) and iron (42.14%). Height for-age and weight-for-height anthropometric indices revealed that many children were located below -1 standard deviation. PMID- 14528361 TI - [The decision to abort: the process and feelings involved]. AB - In Brazil, induced abortion is considered a crime in the majority of cases, the result being that there is little official data on the subject. Little is known about the conditions under which abortions are induced This research was designed to shed light on the characteristics of the women who had had an abortion and to study the reasons why and conditions under which it occurred. The sample consisted of all employees (7,359) and students (2,231) in a university program in S o Paulo who were mailed a self-administered survey. Accompanying the questionnaire was a letter and self-addressed stamped envelope. 27% of the employees and 42% of the students returned the questionnaires. Of these, 1,314 employees and 138 students had had at least one pregnancy. The results presented in this study show that 465 of the employees and students ar some point had thought of having an abortion. They were divided into two groups: those who had had an abortion and those who had nos. The objective was to analyze the association of some characteristics of the women with their decision to have/not have an abortion and how they felt when faced with this decision. The proportion of women who had had an abortion was significantly lower among married women than singles. A larger percentage of women who had talked with a friend and/or husband/partner/boyfriend had decided to have an abortion than those who had talked to a parent or had not talked to anyone. More women who said they were not prepared to raise/educate a child had had an abortion as compared to those giving other reasons. Almost half of the women undergoing an abortion said that they felt bad emotionally and physically afterwards. Among those who had not had an abortion, almost two-fifths reported that they felt good, were happy, relieved, and did not regret their decision. The conclusion drawn from the population studied was that emotional and social factors played a significant role in the decision-making process for women considering an abortion. PMID- 14528362 TI - [A popular concept of schistosomiasis mansoni: modes of transmission and prevention in the perspective of gender differences]. AB - This is a study on popular ideas, beliefs and practices concerning the transmission and prevention of parasitic diseases, specifically schistosomiasis mansoni. The author explored the existence of a popular belief system within the health care system, combining elements from biomedicine and folk medicine for the understanding and management of the health/disease phenomenon. The study describes the differences in perception of morbidity and gender among the interviewees. Finally, the study presents popular discourse on parasitic diseases and discusses difficulties with control measures. It considers the contradiction between cultural values and habits and conventional control measures, critically assessing the lack of precision in health education measures under government prevention programs. PMID- 14528363 TI - [Internal validity in cross-sectional studies: comments based on an investigation on the association between socioeconomic factors and schistosomiasis]. AB - The purpose of this paper was to make some comments on validity based on an investigation developed in an urban area in the Greater Metropolitan Area of Recife, Brazil, the objective of which was to study the association between socioeconomic factors and schistosomiasis. It discusses the distortion in the odds ratio that may be introduced either by the way individuals are selected for the study or by the way the information is collected. The article describes the mechanisms used by the authors to avoid bias when planning or conducting the investigation and considerations were made on how these types of bias could have influenced the results. PMID- 14528364 TI - [Malaria in Brazil: trends in the last ten years]. AB - There have been major changes in the epidemiological patterns for malaria in Brazil in the last ten years. One of the most important features is the tendency towards focal distribution in the Amazon region, instead of spreading transmission zones. This new epidemiological situation has led to changes in the control theory based on the occurrence of focal transmission of the infection. The importance of diversified control measures has thus increased, taking into account the characteristics of each transmission focus. The article comments on new proposals for control presented at the Ministerial Conference in Amsterdam. PMID- 14528365 TI - [Vaccines: a public affair?] [corrected]. AB - In spite of the expectation that the development of vaccines would help in the control of some of the main transmissible diseases in Brazil, which are responsible for a large share of endemic illness in the country, efficient and safe vaccines against no less serious contagious diseases are available, yet underutilized. The reason is simple: some vaccines are not included in the National Immunization Program (PNI) and are this not the object of government campaigns. Despite the competence with which the National Health Foundation has acted-through the PNI-in reducing prevalence of diseases such as diphteria, whooping cough, measles, and polio, lack of information and prejudice make several vaccines inaccessible to a major share of the Brazilian population. Since the government-through its vaccination campaigns-exerts the most important influence on public opinion in this area, a vaccine procedure which is not the object of such campaigns will obviously not become part of popular culture. The result of this lack of information is that the population fails to utilize vaccines against such diseases as rubella, mumps, and hepatitis B or against Haemophilus influenza type b and pneumococcus infections. We therefore argue for information on the existence and availability of these vaccines, so that not only the part of the population that has access to private medicine can benefit, but mainly so that pressure can be brought to bear on the government for all public health care services to supply them to the neediest portion of the population. We suggest that strategies for the socialization of immunoprophylatics should be defined with participation by all segments of society, incorporating measures such as the gradual expansion of the National Immunization Program, coverage of vaccine costs by health insurance policies, and granting of fiscal incentives to companies that vaccinate their employees and families. Finally, we propose a struggle against the prejudice surrounding participation by private initiative in collective health actions, as well as defense of the interaction between private and public sectors in all aspects of health throughout Brazil. PMID- 14528366 TI - [Aids in children: observations on vertical transmission]. AB - This paper seeks to review the epidemiological aspects of the acquired immunodeficiency syndrome (AIDS) in children and the vertical transmission of the human immunodeficiency virus (HIV). The modes of mother-to-infant transmission are discussed, including physiopathological aspects of HIV infection in utero, during, and/or after delivery. PMID- 14528373 TI - [Guidelines for the indications of intracoronary ultrasonography in clinical practice]. PMID- 14528375 TI - Noncholesterol-lowering effects of statins. AB - Statins, 3-hydroxy-3-methylglutaryl coenzyme-A (HMG-CoA) reductase inhibitors, inhibit the rate-limiting enzyme in cholesterol synthesis and lead to a significant reduction of plasma lipid concentrations. As a clear correlation exists between serum cholesterol and cardiovascular risk, statins have become increasingly important in current preventive medicine. Studies prompted by the extraordinary benefits afforded by these drugs have reported minimal changes in the vasculature of hypercholesterolemic patients when compared with clinical benefits and have led to further investigations to determine the underlying reasons for these clinical benefits. The purpose of this review is to present the wide array of systems that HMG-CoA reductase inhibitors are known to influence, which range from adverse events due to coronary artery disease, stroke risk, platelet function, endothelial function, and inflammatory effects to intracellular signaling pathways that control vascular cell migration, proliferation, and differentiation. PMID- 14528376 TI - Carotid artery stenting: current status and future directions. AB - Carotid endarterectomy has been the mainstay of therapy for carotid occlusive disease in stroke prevention. Recent advances in endovascular technology has made carotid angioplasty and stenting a therapeutic modality in the treatment of carotid occlusive disease. The enthusiasm for this minimally invasive carotid intervention is hampered by the possibility of stent-related cerebral embolization, which can result in neurologic complications. Although current available data indicate that the results regarding success and complication rates are similar between carotid stenting and endarterectomy, numerous prospective clinical trials are currently underway to evaluate the efficacy of carotid artery stenting using various forms of cerebral protection devices. This article reviews the current status of carotid stenting, including recent clinical studies and ongoing prospective trials. Strategies relating to cerebral embolization protection are also discussed. PMID- 14528377 TI - Clinical benefit of carotid endarterectomy based on duplex ultrasonography. AB - Carotid endarterectomy has been shown to significantly reduce the risk of stroke caused by carotid artery stenosis in selected patients. Limiting the morbidity and costs of this process without increasing the risks should further improve the benefits of this procedure. Results were prospectively collected from 123 consecutive carotid endarterectomies performed at a community teaching hospital. All patients underwent duplex ultrasonography for preoperative evaluation. Catheter angiography was used on a selective basis. Preferential use of regional anesthetic and selective use of the intensive care unit were applied. The mortality, morbidity, complications, and costs were then compared for the group receiving only preoperative duplex ultrasonography with those undergoing catheter angiography preoperatively. Age, comorbid risk factors, indications for carotid endarterectomy, and incidence of stroke were similar in both patient groups. The rates of mortality, morbidity, and stroke for carotid endarterectomy were low (mortality 0%, morbidity 6.5%, stroke 0.8%). For preoperative evaluation all patients underwent duplex ultrasonography (100%) and 28 (23%) underwent preoperative catheter angiography in addition to duplex ultrasonography. The complication rate associated with catheter angiography was 6/28 (21%). Complications included groin hematoma (7%), pseudoaneurysm (3.6%), bradycardia (7%), and unstable angina (3.6%). Costs for duplex ultrasonography averaged 165 US dollars and additional costs incurred by the use of catheter angiography averaged 4,200 US dollars. Intraoperative assessment of the carotid endarterectomy site did not change based on the use of preoperative catheter angiography. Morbidity, mortality, and stroke rates were the same for the 2 groups. The preoperative use of duplex ultrasonography for the sole evaluation in carotid endarterectomy is well established. The use of preoperative catheter angiography is still preferred by a subset of surgeons. The use of catheter angiography is associated with significant morbidity and additional costs when compared to performing carotid endarterectomy based solely on preoperative duplex ultrasonography. The added costs and morbidity of angiography increase the societal cost of this procedure without significant clinical improvement in patient outcome. PMID- 14528378 TI - "Fast-track" abdominal aortic aneurysm repair. AB - The "open" repair of abdominal aortic aneurysm (AAA) continues to evolve with incorporation of less invasive methods for surgical exposure and the use of patient care pathways for shorter hospital stays. In a consecutive series of 30 patients with infrarenal AAA, a "fast-track" hospital care pathway was implemented that included the following: AAA exposure via a limited (10-15 cm) retroperitoneal incision, use of self-retaining retractor and special vascular clamps/instruments, and prosthetic graft endoaneurysmorrhaphy. Excellent anatomic exposure for graft implantation was achieved with an average operative time of 175 minutes. Use of oral metoclopromide and patient-controlled epidural analgesia resulted in patient ambulation and oral diet on postoperative day 1. Average length of hospital stay was 3.6 days (range: 3-7 days), and no patient required readmission for AAA repair-related or gastrointestinal problems. One patient died (30-day mortality rate of 3.3%) caused by delayed recognition of a splenic injury, and 1 patient sustained an intraoperative ureter injury that was repaired and stented. Although stent-graft exclusion for AAA repair has become popular, the major advantages of endovascular therapy, such as shorter ICU and hospital stays, earlier dietary feeding, and reduction in postoperative morbidity, can also be achieved by using minimal incision exposure for AAA interposition grafting combined with traditional hand-sewn vascular anastomoses. "Fast-track" AAA repair is applicable to the majority of patients with infrarenal AAAs, and vascular surgeons can easily master the technique of "limited" incision retroperitoneal exposure. This approach avoids the concerns of endovascular stent graft durability and the mandatory vascular imaging follow-up to identify endoleak development and AAA enlargement. PMID- 14528379 TI - Upper arm polytetrafluoroethylene grafts for dialysis access. Analysis of two different graft sizes: 6 mm and 6-8 mm. AB - The purpose of this retrospective study was to analyze 2 sizes of expanded polytetrafluoroethylene (PTFE) upper arm grafts for dialysis: 8 millimeters, tapered to 6 mm at the arterial side, and 6 millimeters. All upper arm PTFE grafts (Gore-Tex(R)) were performed between January 1981 and April 1997. Patient characteristics, complication rate, and patency rates were compared for both kind of grafts. Five hundred and seven PTFE grafts were analyzed (183 6-mm grafts and 324 6- to 8-mm grafts). Early failure was found in 5 grafts (2.7%) in 6-mm grafts, and in 5 grafts (1.5%) in 8-mm grafts (not significant). Steal syndrome was found in 1 patient (0.5%) of the 6-mm group, and in 11 (3.4%) of the 8-mm grafts (p=0.085). The rate of late complications requiring surgical repair was 0.56 episode per graft-year in the 6-mm grafts group, and 0.33 in the 8-mm grafts (p<0.001). Primary patency rates of 6-mm grafts were 72%, 33%, and 19% at 1, 3, and 5 years; and secondary patency rates were 86%, 68%, 56%, and 44% at 1, 3, 5, and 6 years, respectively. In the 8-mm grafts group, primary patency rates were 77%, 52%, and 39% at 1, 3, and 5 years; and secondary patency rates were 92%, 84%, 73%, and 66% at 1, 3, 5, and 6 years, respectively. Comparison of patency rates of 6-mm and 8-mm grafts were statistically significant (p<0.001) for both primary and secondary curves. However, secondary survival curves were similar for both kind of grafts in a subpopulation of diabetic patients. The authors conclude that the 8-mm graft, tapered to 6 mm at the arterial side, is a dialysis graft with fewer complications and a better patency rate than grafts of 6 mm placed in the same anatomical position, at least in a population of nondiabetic patients. Steal syndrome was observed in some cases of diabetic and older patients with a large-bore graft. Thus, this kind of prosthesis should be avoided in this population. On the other hand, this is not a prospective, randomized study made with any intention for comparison. Therefore, the aforementioned conclusions must be cautiously considered. PMID- 14528380 TI - The effects of glucose-insulin-potassium solution and BN 52021 in intestinal ischemia-reperfusion injury. AB - The objective of this study was to investigate effects of glucose-insulin potassium (GIK) solution and BN 52021, a platelet-activating factor antagonist, on intestinal ischemia-reperfusion injury. Fifty male Sprague-Dawley rats (200 225 g) were divided into 5 groups each containing 10 rats; group SO, sham operation group; group I, mesenteric ischemia group (for 30 minutes); group R, ischemia plus reperfusion (for 60 minutes); group BR, ischemia-reperfusion plus BN 52021; group GR, ischemia-reperfusion plus GIK solution. Samples for malondialdehyde (MDA) and ileum (for mucosal injury score) were obtained. The mucosal injury scores of group R were significantly higher than those of group I (4 +/-0.20 and 3 +/-0.16, respectively, p<0.0001). The scores of groups BR and GR were significantly lower than those of group R (p<0.0001 and p<0.0001, respectively). When it was compared with the injuries in BR and GR groups, similar results were obtained in both groups (p=0.190). Mean MDA levels of group R were significantly higher than those of group I, BR and GR (131.33 +/-3.99 nmol/g, 93.74 +/-3.22 nmol/g, 104.81 +/-2.56 and 100.34 +/-5.30, respectively, p<0.0001). MDA levels of group BR and GR were significantly lower than those of group I (p<0.0001 and p=0.003, respectively). These observations suggest that treatment with GIK solution and BN 52021 before reperfusion and during reperfusion period may be useful in decreasing intestinal reperfusion injury. PMID- 14528381 TI - Noninfectious saccular abdominal aortic aneurysm--a report of two cases. AB - Saccular aortic aneurysms are eccentrically shaped and are commonly thought to be the consequence of infection of the aorta. However, saccular-type aneurysms can be found independent of any clinical or subclinical infection and can be associated with a penetrating aortic ulcer. Moreover, penetrating aortic ulcers have been described in the thoracic aorta, but there is a paucity of any documentation of this clinical and pathologic entity occurring in the abdominal aorta. The authors describe 2 cases of symptomatic, penetrating atherosclerotic ulcers of the abdominal aorta causing saccular, noninfectious abdominal aortic aneurysms. PMID- 14528382 TI - Diagnostic options in assessing abdominal aortic endograft infection--a case report. AB - Endovascular treatment of aortic aneurysms has gained widespread popularity in recent years. Stent grafts have emerged as another option in the surgeon's armamentarium in the treatment of aneurysmal disease. The infectivity of endovascular grafts and therapy for associated graft infections is unknown. Aortic graft infections have the potential for disastrous complications. This report presents a 72-year-old woman with persistent fever and an infected aortic stent graft in the early postoperative period. PMID- 14528383 TI - 18-fluorodeoxyglucose positron emission tomography as a novel imaging tool for the diagnosis of aortoenteric fistula and aortic graft infection--a case report. AB - The diagnosis of aortic graft infection and aortoenteric fistula can be difficult to establish using conventional radiographic imaging modalities. Positron emission tomography (PET) imaging with 18-fluorodeoxyglucose (FDG) can rapidly provide anatomically clear images and define areas of inflammation with increased glucose metabolism. In this report the authors present a case of aortoenteric fistula diagnosed by FDG-PET. Early diagnosis led to rapid surgical intervention with graft removal and extraanatomic bypass. These encouraging results warrant larger controlled studies to evaluate the utility of FDG-PET in the diagnosis of prosthetic aortic graft infection. PMID- 14528384 TI - Treatment of a celiac artery aneurysm with endovascular stent grafting--a case report. AB - As a result of more sophisticated and more commonly performed investigative procedures, aneurysms of the visceral abdominal vasculature, including celiac artery aneurysms, are increasingly recognized. Traditional therapy for visceral artery aneurysms has been limited to open aneurysmectomy or aneurysmorrhaphy to prevent catastrophic aneurysmal rupture. However, these procedures are associated with significant postoperative morbidity and mortality despite technical successes. High complication rates are likely related to poor preoperative conditions among the patient population typically presenting with these visceral artery aneurysms. This report introduces an alternative therapy for visceral artery aneurysms and highlights the potential for catheter-based interventions. This case report depicts a 61-year-old morbidly obese woman diagnosed with a 10 centimeter celiac artery aneurysm during investigation of upper abdominal pain. Given the patient's poor medical condition, punctuated by hemodynamic instability, open operation was avoided, and percutaneous embolization was not feasible owing to a large aneurysm neck. Therefore, inflow to the celiac artery aneurysm was excluded by placing a modular stent graft component within the abdominal aorta at the celiac artery orifice. During the intervening 12 months since stent graft deployment, the aneurysm sac diameter has steadily decreased, as determined by serial computed tomography scans. This report underscores the potential for catheter-based techniques to offer new therapeutic options for patients with visceral artery aneurysms. Careful individualization is required given the highly variable size, location, and character of such lesions. PMID- 14528386 TI - Evaluation of post-application rheological changes in cosmetics using a novel measuring device: relationship to sensory evaluation. PMID- 14528385 TI - Pseudoaneurysm of the aortic arch after aortosubclavian bypass treated with endoluminal stent grafting--a case report. AB - This paper describes a case of aortic arch pseudoaneurysm treated with stent graft 2 years after aortosubclavian bypass repair of a subclavian artery aneurysm. An 84-year-old man presented with back pain. Two years before, he had had a left subclavian artery aneurysm repaired with aortosubclavian bypass. Upon examination by computed tomography (CT) scan and angiography he was found to have a bovine arch configuration, a 7-centimeter pseudoaneurysm arising from the stump f the native subclavian artery, a patent aortosubclavian bypass, and a left hemothorax. A 37 mm by 10 mm Gore Excluder thoracic graft was introduced into the right femoral artery cutdown and deployed across the arch, excluding the pseudoaneurysm and preserving the brachiocephalic vessels. Follow up CT scan at 1 year shows exclusion of the pseudoaneurysm. The patient continues to do well 1 year after implantation without evidence of endoleak. In the presence of unusual anatomical characteristics, endoluminal stent graft repair can be successfully performed across the aortic arch. PMID- 14528388 TI - Influence of internal structures of hair fiber on hair appearance. III. Generation of light-scattering factors in hair cuticles and the influence on hair shine. AB - The effects of thermal treatments on hair fiber induced by blow-drying have been investigated. It was found that the hair shows whitish and powdery appearance after heat drying, especially when dark hair is rapidly dried from a wet condition. For all kinds of hair, the appearance of numerous glittering speckles was confirmed on the cuticle surface by optical microscopic observations. SEM images of hair transverse and longitudinal sections with glittering speckles revealed that the splitting of cuticle layers generated by blow-drying occurred not only at the outermost parts of cuticle cells but also at the inner parts of the cellular interfaces. The release and uptake of moisture through fiber surfaces induces deformation of cuticle cells, probably because of anisotropic swelling or drying of the cells. The cuticles with glittering speckles are found to be fragile and are easily damaged in combination with other mechanical stresses such as combing force. Furthermore, the authors have found an efficient system for both improving hair shine and preventing cuticle damage caused by the blow-drying/combing process. PMID- 14528387 TI - Optical properties of hair: effect of treatments on luster as quantified by image analysis. AB - Image analysis has been employed to measure the luster of hair simulated by light reflected from a curved hair tress. Hair samples (up to four) were mounted side by-side in a special sample holder in the form of a cylinder and illuminated by a uniform beam of white light. Digital images of hair tresses were captured with a high-resolution camera and were analyzed by scanning across highlighted and dark areas of the resultant image using image analysis software with developed macros. Plots, similar to goniophotometric scattering curves, were used to calculate luster values according to previously published work by Nickerson, Stamm, and Reich-Robbins. Both the Stamm and Reich-Robbins approaches were found to give similar results, while the Nickerson gloss parameter exhibited less sensitivity to hair modification with cosmetic ingredients. The procedure was employed to assess the luster of natural white, light blonde, light brown, medium brown, and dark brown hair, and revealed an increase in luster indices in proportion to an increase in fiber pigmentation. Cosmetic oils such as phenyl trimethicone, amodimethicone, and castor oil were also found to increase the luster of hair as a result of the change in contrast between specular and diffuse reflection. Styling resins such as butyl ester of PVM/MA copolymer, vinyl caprolactam/PVP/dimethylaminoethyl methacrylate copolymer, and isobutylene/ethylmaleimide/hydroxyethylmaleimide copolymer were shown to increase hair gloss by a similar mechanism, as evidenced by calculated higher values of the Stamm and Reich-Robbins luster parameters. An effect of hair dulling by deposition of micronized ZnO at various concentrations, as well as by synthetic sebum, is also discussed. PMID- 14528389 TI - Scalp hair length. II. Estimating the percentages of adults in the USA and larger populations by hair length. AB - Scalp hair length assessments by anatomical site, previously made in Florida theme parks on adults, are related to anatomical measurements to obtain estimates of free-hanging hair lengths in centimeters. A plot of the natural logarithm of the percent population versus these hair lengths provides a straight line and an equation that permits the estimation of the numbers of persons in the USA and larger populations with hair lengths up to 183 cm (just beyond ankle-length). Data were also collected via a literature search for even longer hair lengths (ankle-length or longer) to provide an equation to estimate the numbers of persons with exceptionally long hair. A comparative plot of these two equations suggests that "normal" anagen periods may be considerably longer than current estimates in the literature. PMID- 14528390 TI - Chemical and photo-oxidative hair damage studied by dye diffusion and electrophoresis. AB - Microspectrophotometric and electrophoretic methods were used to characterize and quantify the effects of primary damage to hair from chemical and photochemical oxidative processes. The diffusion of molecules proceeding from the fiber surface to the center of untreated and modified (by chemical and photochemical oxidative processes) hair fibers was mapped by fluorescence microscopy and quantified by calculating diffusion coefficients of a fluorescent molecule. In addition, an electrophoretic separation technique, namely, SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis), was used not only to substantiate the results obtained in the microfluorometric study, but also to show how the main classes of proteins of unaltered hair are modified by cosmetic chemical treatments, light exposure, and combinations of these two processes. UV microspectrophotometry is an alternate analytical method to evaluate photo oxidative damage in hair, and supports the results obtained by microfluorometry. PMID- 14528391 TI - New insights into the physicochemical effects of ammonia/peroxide bleaching of hair and Sepia melanins. AB - Chemically unaltered melanosomes from black hair were isolated using a mild enzymatic procedure reported by Novellino et al. involving sequential treatment of a homogenized hair sample with different protease enzymes. Time-dependent fluorescence studies show, under identical conditions, that the rate of bleaching upon NH3/H2O2 treatment of hair melanosomes is twice that of Sepia melanosomes. The structure and morphology of hair melanosomes are compared to Sepia eumelanin using ESEM and TEM imaging studies. Black hair melanosomes are aggregates of rice shaped ellipsoidal particles (0.8-1.0 microm in length and 0.2-0.6 microm in width) surrounded by an amorphous material suspected to be made of non proteinacious materials. Sepia eumelanin aggregates are larger (2-5 microm) particles with a "doughnut" shape comprised of 100-150-nm spherical particles. Time-dependent TEM imaging studies of ammonia-treated (pH 10) hair melanosomes showed an initial breakdown of melanosomal aggregates followed by rupture of the melanosomal membrane, releasing melanin nanoparticles and leaving a ghost membrane behind. After prolonged treatment with aqueous NH3, a total loss of characteristic melanosome morphology was observed leading to an amorphous material. By contrast, Sepia melanosomes under identical conditions of ammonia treatment did not show such changes, probably due to different surface properties and aggregation behavior. Sodium hydroxide or sodium carbonate at identical pH did not show similar changes to ammonia, suggesting that the changes are not merely due to alkaline pH, but, rather, are specific to ammonia. Co-treatment with ammonia and peroxide induced a faster disintegration of the melanosomes, resulting in a complete dissolution and discoloration of melanin in 30 minutes. The data suggest that ammonia helps to release melanin nanoparticles out of melanosomes, making them more susceptible to oxidative attack by H2O2. PMID- 14528392 TI - The preservative efficacy testing method for powdered eye shadows. AB - Preservative efficacy testing is based on a sample inoculation using a microbial suspension with a determined amount of colony-forming units (CFU). After that, the number of survivors is investigated by periodic evaluations, and the results are compared with specifications. When liquid cosmetics are evaluated, it is easy to obtain homogeneity between the inoculum and the sample, but for a powder sample it cannot be guaranteed. In this context, freeze-dried microorganisms could be used to help the homogenization. In this research, the preservative efficacy is evaluated using a powdered eye shadow. The microorganisms used were Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, and Aspergillus niger. The challenge tests were performed in samples with (P) or without (NP) added preservative. The methods used to evaluate the results were the ones described in the official compendia and in the cosmetics guides of international associations, also using linear regression in calculating the D-value. The results showed that it is possible to use freeze-dried microorganisms instead of suspension to evaluate the preservative efficacy of cosmetic solids. The microorganism stability was verified by the determination of the microbial load and the minimum inhibitory concentration after freeze-drying and during the following six months. PMID- 14528393 TI - Optimization of in vitro skin permeation by lactic acid from gel formulations. AB - The purpose of this study was to evaluate the effect of the concentration as well as the vehicle's pH on in vitro skin permeation by lactic acid from gel formulations, using an optimization technique. Nine gels containing 3%, 6%, and 9% (w/w) lactic acid in three different phosphate buffers, with pH values of 2.8, 3.8, and 4.8, were prepared and were applied in modified Franz diffusion cells. The pH of the vehicles and the lactic acid concentrations were used to create a mathematical model that correlates these factors with the cumulative amount of lactic acid permeated through human cadaver epidermis. For this purpose, the optimization technique 3(2) was applied. It was found that the correlation of the above factors can be adequately described with a polynomial equation, which can be used for predicting the cumulative amounts of lactic acid permeated. The results indicated that as the lactic acid's concentration increased, the cumulative amount permeated also increased after 24 h at all pH values. Moreover, the amount of lactic acid permeated decreased as the pH of the gels was increased. The greater amount permeated at all time intervals (6 h, 9 h, 12 h, and 24 h) was obtained when the concentration of lactic acid was 9% and the pH of the gel formulation was 2.8. PMID- 14528395 TI - A case of pregnancy complicated by the development of a tubo-ovarian abscess following in vitro fertilization and embryo transfer. AB - Tubo-ovarian abscess is an uncommon complication in pregnant women. In this report, we present a patient who developed a tubo-ovarian abscess during pregnancy following in vitro fertilization and embryo transfer (IVF-ET). Despite treatment with intravenous antibiotics she delivered at 22 weeks of gestation. After delivery, she underwent a left salpingo-oophorectomy. Review of the literature revealed only 26 reported cases of tubo-ovarian or pelvic abscess during pregnancy. Pelvic inflammatory disease, previous laparotomy, and structural genital anomalies are known risk factors for pelvic abscess during pregnancy. Pelvic abscess resulting as a complication of vaginal oocyte retrieval has been reported. Therefore, although enabling women with organic pelvic disease such as endometriosis and hydrosalpinx to achieve pregnancy, assisted reproductive techniques may potentially result in pelvic infection during pregnancy. This case suggests that a preconception evaluation and treatment for such conditions should be considered for women undergoing treatment for infertility. PMID- 14528396 TI - Indicative markers of cell proliferation and apoptosis during the perinatal period. AB - Tissue polypeptide-specific antigen (TPS), an indicative marker of cell proliferation and soluble Fas (sFas), an antiapoptotic molecule were determined in neonatal serum (day 1-N1 and day 4-N4 of life), compared with maternal serum (MS) and umbilical cord serum (UC) to study changes of these markers during the perinatal period. Serum TPS and sFas concentrations were measured in 33 healthy, termed neonates, their mothers and 25 healthy nonpregnant controls (CS), age matched to the mothers. TPS serum concentrations were significantly elevated in N1 and N4 as compared to CS (p < 0.0001 and p < 0.0003), increasing significantly from UC to N1 (p < 0.0001) and decreasing thereafter in N4 (p < 0.0002). MS serum concentrations, being significantly higher than CS (p < 0.0001), UC (p < 0.0001) and N4 (p < 0.003), but lower than N1 (p < 0.02) were strongly depended on the mode of delivery (p < 0.001). Serum concentrations of sFas, being lower in UC than in MS or CS (p < 0.0001), increased significantly in N4 samples (p < 0.01). A strong correlation was found between sFas serum concentrations in N1 and N4 (r = 0.65; p < 0.001). Our results demonstrate significant perinatal changes in TPS and sFas serum concentrations, possibly indicating gradual decrease of proliferation and apoptosis in early postnatal life. PMID- 14528397 TI - Brachial plexus paresis associated with fetal neck compression from forceps. AB - Instrumental vaginal deliveries have been associated with higher risks of brachial plexus injuries. The proposed mechanisms involve the indirect association of instrumental deliveries with shoulder dystocia and nerve stretch injuries secondary to rotations of 90 degrees or more. We present a brachial plexus paresis resulting from direct compression of the forceps blade in the fetal neck. A term infant was delivered by a low Kielland forceps rotation. No shoulder dystocia was noted. The immediate neonatal exam revealed an Erb's palsy and an ipsilateral bruise in the lateral aspect of the neck. The paresis resolved during the first day of life. Direct cervical compression of the fetal neck by forceps in procedures involving rotations of the presentation may result in brachial plexus injuries. PMID- 14528398 TI - Antenatal imaging of a near-term fetus with primary acalvaria. AB - We report a near-term fetus with primary acalvaria, an extremely rare malformation characterized by the complete absence of the membranous neurocranium. Sonographic findings were considered consistent with this condition, which was confirmed by magnetic resonance imaging. Spontaneous vaginal delivery resulted in a fresh stillbirth, with an intact cranial sac. PMID- 14528399 TI - Echocardiographic assessment of preload conditions does not help at the neonatal intensive care unit. AB - To determine the value of noninvasive assessment of right ventricular preload in neonates, a prospective unblinded study was performed. Thirty-seven neonates without heart disease (median birth weight 1390 g, range 900 to 4400) were studied at the neonatal intensive care unit, comparing directly measured central venous pressure (CVP) and two-dimensional echocardiographic measurement of the maximum and minimum diameter of the inferior vena cava and calculated vena cava index (VCI). CVP was higher in conventionally ventilated and high-frequency oscillatory ventilated neonates than in those breathing spontaneously ( p < 0.0001). VCI in high-frequency oscillatory ventilated patients was lower (5 +/- 4) than in spontaneously breathing (56 +/- 19) and conventionally ventilated (49 +/- 19) (p = 0.002) neonates. CVP and VCI were inversely correlated in spontaneously breathing (r = -0.631), but not in conventionally and high frequency oscillatory ventilated patients. VCI does not predict CVP in ventilated premature neonates, the correlation is limited only to spontaneously breathing infants. PMID- 14528400 TI - Obstetric outcomes of human immunodeficiency virus-1-serodiscordant couples following in vitro fertilization with intracytoplasmic sperm injection. AB - The objective of this article is to report obstetric outcomes of human immunodeficiency virus-1 (HIV-1)-serodiscordant couples who underwent in vitro fertilization and embryo transfer (IVF-ET) with intracytoplasmic sperm injection (ICSI) at a tertiary care center. We reviewed the outcomes of seronegative women after IVF-ET with ICSI from January 1, 1997 to June 1, 2002. Serodiscordant couples (n = 25) successfully conceived 27 pregnancies delivering 40 neonates (16 singletons, 9 twins, and 2 triplets). The mean gestational age at delivery was 37 0/7 weeks +/- 3 6/7 weeks (range 26 0/7 to 41 2/7 weeks). The mean birth weight was 2646 g +/- 952 g (range 678 to 4396 g). The cesarean section rate was 70%. Preterm delivery (<37 weeks) occurred in 7 pregnancies, and low birth weight (<2500 g) was observed in 8 pregnancies. There were no HIV-1 seroconversions detected at delivery. One hundred percent of the mothers and offspring were beyond 3 months postpartum and remained seronegative. IVF-ET with ICSI seems safe and effective for serodiscordant couples. Obstetric outcomes are favorable, and HIV-1 infection risk is limited. PMID- 14528401 TI - Risk factors predicting vasopressor use after patent ductus arteriosus ligation. AB - Ligation of the patent ductus arteriosus (PDA) is sometimes complicated by postoperative hypotension requiring vasopressor(s). It is unclear which infants are at risk for this complication. We conducted a retrospective and prospective cohort study to identify risk factors predicting vasopressor use after PDA ligation. Our patients were infants < 37 weeks of gestation who underwent PDA ligation. The primary outcome was the use of vasopressor(s) within 72 hours after PDA ligation, defined as beginning vasopressor(s) or increasing the dose of vasopressor(s). Thirty-two of 100 (32%) study infants required vasopressor(s) after PDA ligation. Infants who had lower birth weights, lower gestational ages, higher ventilator support, or whose mothers had received antenatal steroids had a higher risk of vasopressor use. Infants who required vasopressor(s) were at increased risk of dying before 36 weeks postmenstrual age. Lower birth weight, lower gestational age, and higher respiratory support define a high-risk subgroup of patients in whom the prophylactic administration of vasopressor(s) could be studied. PMID- 14528402 TI - A population-based study of the effects of birth weight on early developmental delay or disability in children. AB - Improving medical treatment of extremely low-birth-weight infants over the last 20 to 30 years resulted in increased survival rates. The developmental sequela of salvaged infants is of great interest to perinatologists. The primary purposes of the current study were to assess the effect of birth weight (BW) on developmental delay or disability (DDD) in the first three years of life and determine whether there is a BW threshold below which all infants should be evaluated to determine if intervention services for children with DDD should be received. Three statewide databases were merged: 1998 Birth Vital Statistics; 1997-1998 Medicaid eligibility files; and 1998-2001 Children's Medical Services' Early Intervention Program (CMS-EIP) data. Infants who died within the first year of life and plural births were excluded. The final dataset consisted of 170,874 records. A child was determined to have a DDD if a developmental delay, or an established condition, such as sensory impairment, genetic, metabolic, neurological, or severe attachment disorders, was diagnosed through a multidisciplinary evaluation. Logistic regression models were used to relate BW to DDD, controlling for sociodemographic, behavioral, and perinatal variables. Adjusted odds ratios (OR) were calculated to describe the effects of BW on DDD. There was a significant effect of BW on DDD (Adjusted OR &equals 97.50, 40.01, 15.84, 3.29, 1.39, 1.00, 1.52 for BW categories 450-749, 750-999, 1000-1499, 1500- 2499, 2500-2999, 3000 4749, 4750-6050 g, respectively). In these categories, 70%, 56%, 36%, 11%, 4%, 3%, and 6% of surviving singleton infants, respectively, suffered a DDD in their first 3 years of life. Four medical, five sociodemographic, and two behavioral factors were significant in addition to BW. An equation for predicting the probability of DDD given these factors was obtained, and its use exemplified. BW is strongly associated with DDD. Over 60% of infants weighing < 1000 g and nearly half (46%) of those weighing < 1500 g at birth are diagnosed with a DDD before 3 years of age. The probability of DDD for a specific infant also varies by sociodemographic, other perinatal, and behavioral factors. The results of this paper suggest that all surviving infants of BW < 1000 g, and perhaps < 1500 g, should be automatically referred for evaluation. PMID- 14528403 TI - Are perinatal risk factors helpful in predicting and optimizing treatment strategies for transient hypothyroxinemia in very-low-birth-weight infants? AB - Transient hypothyroxinemia is common in premature infants and has been associated with intraventricular hemorrhage (IVH), periventricular leukomalacia (PVL), poor neurodevelopmental outcomes, and mortality. Recent trials have failed to show that supplemental thyroid hormone improves overall neurodevelopmental outcome. The objective of this article is too determine perinatal risk factors for transient hypothyroxinemia (TH). We studied a cohort of infants born between July 1993 and July 2000 who were less than 1500 g and who received a newborn screening for thyroid function ( n = 932). Total serum thyroxine (T(4)) was collected routinely on the fifth day of life. T (4) was correlated with gestational age (R = 0.59, p < 0.01). After controlling for potential confounding variables, gestational age, dopamine, and mechanical ventilation were found to be independently associated with low T (4) (overall model: r(2) = 0.41, p < 0.01). Number needed to treat (NNT) analysis showed treating all infants less than 27 weeks would lead to treating 6.3 infants for every one with a subsequent T(4) < 5 microg/dL. By combining gestational age and need for dopamine support, NNT = 2.4 for every one infant with subsequent T(4) < 5 microg/dL. Low gestational age, mechanical ventilation, and need for dopamine were associated with low T(4) levels and may be helpful in optimizing treatment strategies for TH. PMID- 14528404 TI - [Empirically validated therapists instead of empirically validated psychotherapy?]. PMID- 14528405 TI - [Psychosocial screening and selection of candidates for liver transplantation]. AB - 281 consecutive candidates for liver transplantation were referred to our consultation-liaison service for psychiatric evaluation over a 4-year period. For these candidates a semistructured psychiatric interview according to the codes listed in the DSM-III-R and the Transplant Evaluation Rating Scale (TERS) classifying patients' level of adjustment in psychosocial functioning were completed. Our study showed a prevalence rate of psychiatric disorders of 65.8 % among candidates. The categories of alcohol abuse (27.8 %) and alcohol dependence (11.7 %) were the most frequent psychiatric diagnoses. The mean TERS summary score of the whole sample was 33.38 +/- 7,31. 68.4 % of the sample were considered good candidates, 18.1 % of the candidates were identified borderline, but acceptable under some conditions, and 13.5 % of the referrals were rated poor liver transplant candidates. The TERS summary scores discriminated well between the three rating categories of candidate quality. Apart from a careful assessment of psychiatric morbidity including substance abuse disorders, the C-L psychiatrist should always rate psychosocial functioning referring to coping, compliance, social support and lifestyle factors among liver transplant candidates. The TERS therefore may be used routinely. PMID- 14528406 TI - [The relevance of zen-buddhism for dialectic-behavioral therapy]. AB - Dialectic-Behavioral Therapy is a specific psychotherapeutic approach to answer the needs of patients with Borderline Personality Disorder. It uses concepts and techniques of Cognitive Behavioral Therapy and of Humanistic Psychotherapies. For a deeper understanding, it is necessary to include also its Zen-Buddhistic background. The experience of Zen-meditation and the basic philosophy of Zen Buddhism will be explained. In the context of the historical relation between Zen Buddhism and Psychotherapy, the position of the DBT will be specified. Finally it will be demonstrated how Zen-Buddhism inspired the practice of DBT and what kinds of problems arise when a modern psychotherapy uses the concept of a premodern conception of the world and human existence. PMID- 14528407 TI - [Effectiveness of outpatient treatment for alcoholism - impact of personality disorders on course of treatment]. AB - Personality disorders (PS) frequently exist as comorbid disorder in alcoholism. However, the impact of comorbid personality disorder on the treatment and course of alcoholism still remains unclear. On that background, the present study on the efficacy of an intensive out-patient therapy for alcohol-dependents investigated the influence of comorbid personality disorder on the relapse and dropout rate during the treatment phase in 102 patients. Personality disorders were assessed with the SCID-II (DSM-IV). On a descriptive level results indicate that comorbidity with negativistic personality disorder and Cluster B PS may adversely affect the course of treatment. Statistical analyses, however, revealed that the observed differences in the type or number of personality disorders between patients with or without relapse resp. between patients with or without dropout failed to reach statistical significance. Thus, we conclude that the relevance of comorbid personality disorder on the course of alcoholism may be overestimated. The present results indicate that patients with comorbid personality disorder can be successfully integrated into a high-structured outpatient therapy. PMID- 14528408 TI - [Improving medical students' interactive competence]. AB - We report a new curriculum for improving medical students' interactive competence. The curriculum is based on the method of "psychotherapeutic plan analysis" by F. Caspar in combination with elements of systemic supervision and problem-based learning. For evaluation, participants (fifth year medical students, n = 50) were asked to fill in a questionnaire adapted to the contents of the course. Comparison was made possible by a control group (n = 72) teached in a traditional manner. Evaluation showed significant improvement of the students' self awareness, perception of their patients and sensibility for patients' needs. The students expanded their capability to reflect own actions and to work in teams. Initially expressed anxiety concerning contact with patients was reduced significantly during the course. In three and twelve months follow-ups, students still considered their personal achievement to be significantly high. Participants of the curriculum rated their courses higher with regard to didactic issues, improvement of interactive competences and personal relevance, while control courses were seen to be more relevant to prepare for exams. PMID- 14528409 TI - [New bridges for gaps in psychotherapeutic service provision by the internet: hopes, challenges and a solution]. AB - Modern communication technologies such as internet, e-mail or SMS open up new possibilities for the optimization of the psychotherapeutic service provision. However, their use places new challenges. Ethical and legal requirements must be considered, technical and communicative aspects must be integrated into supplying concepts. This paper presents central requirements for the use of the Internet and a possible solution by the example of the project "Internet-Brucke" (internet bridge). In this project internet chat groups are tested as an aftercare program following inpatient treatment in a specialized hospital for psychotherapy and psychosomatic medicine. It is discussed, in what respect the solutions fit the specific requirements of this project and what can be learned for further applications. PMID- 14528410 TI - [The University of Rhode Island Change Assessment Scale (URICA)]. AB - The present paper describes a German version of the University of Rhode Island Change Assessment Scale (URICA) called Fragebogen zu Erfassung der Veranderungsbereitschaft (FEVER). The URICA is a questionnaire designed to measure the stages of change. This instrument is well suited for complex problem behavior, because it yields scores for each stage of change for each individual instead of classifying individuals into a single stage. It is based on Prochaska's transtheoretical model of change. This model recognizes that most psychological disorders are characterized by repetitive problem behaviors that are resistant to modification, and that people do not make a simple black or white decision to change their behavior, rather it holds that people change their behavior gradually across different stages. The model has been successfully applied in different areas of health care to help preventive and treatment interventions adjust to clients' readiness for change. The FEVER was used and tested in a cross-sectional study of 129 patients with an eating disorder. It turned out to be an instrument that was easily understood by patients and that yielded the factor structure of the original version. The internal consistency of the scales was good or very good. Means and standard deviations of the scales were comparable to those found using the original instrument. In addition, there was evidence for the predictive and concordant validity of the instrument. We conclude that the FEVER is a valid instrument to assess the motivational dimension of therapeutic change of complex problem behavior. PMID- 14528411 TI - [Did the Diagnostic Profile in Psychosomatically Examined Patients Seen by Consultation/Liaison-Services (CL Services) of Berlin Hospitals Change?]. AB - The aim of this study was to find out whether the diagnostic profile for patients seen in the psychosomatic Consultation-Liaison (CL) services in three Berlin hospitals in 1994 and 2001changed. All referrals to the psychosomatic CL service within 3 months in 1994 (N = 229) and within 3 months in 2001 (N = 196) were recorded and documented. It could be shown that the referred patients of the three Berlin hospitals more frequently had only mentally caused symptoms (without organ diseases) in the first data recording (1994) and more frequently organic diseases with concomitant mental comorbidity in the second data recording (2001). If this finding reflects a general tendency, it requires an adjustment in the competence of psychosomatic CL services and health policies that ensure the inclusion of mental aspects in hospital care. PMID- 14528412 TI - [The adult attachment projective (AAP) - psychometric properties and new reserach results]. AB - George, West and Pettem developed a new measure, the Adult Attachment Projective (AAP) to assess attachment representation in adults. The AAP is comprised of a set of eight drawings, one neutral scene and seven scenes of attachment situations. Although the pictures were drawn as projective stimuli, the method of administration combines projective and interview techniques in the form of a semi structured interview. In this paper the coding procedure and attachment classifications of the AAP will shortly be described. The current results on reliability and convergent validity are reported. Developmental studies examining correlates of attachment during the preschool-age years, as well as adult attachment classification and foster mothers' perceptions of their relationship with their at risk foster children give first evidence for the predicitve validity of the AAP. The results of a recent study with dysthymic women using the AAP add to the increasing number of studies that have identified an association between preoccupied attachment and depression. This study encourages the use of the AAP in a broader clinical context. The modified application of the AAP using functional MRI opens a new approach to assess neural correlates of attachment representation in patients with a Borderline Personality Disorder compared to controls, and other clinical groups. PMID- 14528413 TI - [Quantitative CTG appraisal sub partu with a new CTG score: diagnostic significance of the parameters of the acid-base balance in umbilical blood?]. AB - BACKGROUND: As a role, the severity of "CTG pathology" in acute or chronic fetal hypoxia could only be registered but not quantified exactly by visual examination. For this reason, the term "CTG pathology" has not been unequivocally defined. The objective of the present study was to describe the dependence of fetal heart rate phenomena on the parameters of the acid-base balance quantitatively and to subject the latter to statistical analysis. METHODS: The last 120 minutes of a total of 411 interference-free cardiotocograms directly registered intra partum (HP, 2 cm/min) were evaluated in terms of the visual appearance of CTG: the frequency level and the fluctuation of every single minute and the deceleration-area sum were considered for 30 CTG minutes per fetus. A new CTG score was conceived which envisages six points for the mean frequency according to specific definitions, three points for the deceleration-area and further six points for the fluctuation (long-term variability). The heart rate data and clinical factors were analyzed off-line using parametric and non parametric test procedures with special programs (FORTRAN) on an IBM system (6150). Besides the pH values in the umbilical arterial and venous blood, direct measurements of blood gases (pO(2), pCO(2)) were also available in 113 cases, so that the base excess (mmol/l) and the oxygen saturation (%) could be calculated according to known algorithms. RESULTS: For the last 30 minutes ante partum, the CTG score correlates highly significantly (P << 10(-4)) with the actual pH value in the umbilical arterial blood (r = -0.6002). The correlation with the base excess is better (r = 0.642, P << 10(-4)), whereas the correlation with the directly measured pCO (2) is weaker (r = 0.587). Such strict correlations could not be demonstrated for oxygen saturation in the umbilical arterial or umbilical venous blood (r = -0.258, P = 0.0057 and r = -0.308, P = 0.0009). CONCLUSIONS: The good correlations of the variables of the fetal acid-base balance with the CTG phenomena registered with the score can be used for clinical purposes. However, analysis of the CTG with the naked eye is not practically feasible for the obstetricians; it must therefore be done per computer. Quantitative CTG analysis has created the perspective of a promising new method for fetal monitoring sub partu. PMID- 14528414 TI - [Evaluation of the reproducibility of Doppler ultrasonographic measurements in obstetrics]. AB - BACKGROUND: Are obstetric doppler ultrasonographic measurements of the fetal and maternal flow parameters reproducible? PATIENTS AND METHODS: For internal quality management, doppler ultrasonographic measurements were performed on 81 patients (random screening sample) with an Acuson Sequoia Ultrasound at the Universitatsfrauenklinik Marburg. Successively two experienced investigators measured the umbilical artery, medial cerebral artery, and the uterine arteries. The correlation between the measurements of the two investigators was presented in a spread chart. In order to exclude systematic differences between the measurements, linear regression was analyzed and the distance to the abscissa was calculated. A relative divergence of more than 20% was determined as a non corresponding measurement. RESULTS: The distance of the linear regression to the abscissa was calculated for the four vessels: umbilical artery 0.4 (95% CI; 0.124 - 0.486), medial cerebral artery 0.9 (95% CI; 0.534 - 1.264), right uterine artery 0.2 (95% CI; 0.124 - 0.305), and left uterine artery 0.2 (95% CI; 0.121 - 0.317). Concerning the four arteries, a divergence of more than 20% between the two investigators was found: umbilical artery 16%, medial cerebral artery 42%, right uterine artery 28%, and left uterine artery 37%. CONCLUSIONS: Because the interobserver variability was surprisingly high and acceptable correlation could be stated only for the umbilical artery, internal quality standards are essential. Routine use of different devices should be made after careful consideration only, especially if clinical decisions are to be based on them. PMID- 14528415 TI - [Circadian blood pressure rhythm in preeclampsia as a predictor of maternal and obstetrical outcome]. AB - BACKGROUND: Preeclampsia is a pregnancy-induced multiorganic disease. The incidence is 5 - 7 % in all pregnant women. To assess the value of circadian blood pressure rhythm in preeclamptic women, 24-hour blood pressure monitoring might be useful. The purpose of this study was to determine the predictive value of circadian blood pressure rhythm for the maternal and obstetrical outcome in patients with mild preeclampsia. PATIENTS AND METHODS: Sixty-six patients with mild preeclampsia underwent 24-hour blood pressure monitoring between 29 and 39 weeks gestation. Twenty-eight patients with normal circadian blood pressure rhythm were compared with thirty-eight patients with suspended circadian blood rhythm. RESULTS: There was no difference in age, body mass index, gravidity, parity, miscarriage, or fetal outcome between the two groups. The rate of secondary caesarean section was 39.5 % in the suspended circadian blood pressure group and 21.4 % in the normal circadian blood pressure group, respectively (P = 0.03). CONCLUSIONS: In patients with mild preeclampsia, the circadian blood pressure rhythm appears to be unsuitable as a screening test. Further research is required for more etiological and pathophysiological insights into the development of clinically useful tools with predictive value. PMID- 14528416 TI - [Why are babies born at night at increased risk of early neonatal mortality?]. AB - BACKGROUND: Increased perinatal and neonatal mortality rates have been previously reported in night-time births compared with births during the day. This effect has been attributed to decreased quality of medical care during the night. However, alternative explanations exist such as decreased birth-weight of night births. The objective of this study was to further investigate this relationship. MATERIALS AND METHODS: Data from 590,332 low risk births (singleton births, > or = 2500 g birth-weight, no major congenital anomaly) were obtained from the perinatal birth register of Hesse, Germany, 1990-2000. Outcome was defined as either death during labour or within 7 days of life. Night-time births were defined as births between 9.00 p.m. and 6.59 a.m., otherwise day-time births were assumed. Subgroup analyses and logistic regression analyses were performed to assess whether the excess mortality of night-births might be explained by other factors. RESULTS: Mortality rates were increased in night-time births (RR = 1.26; 95% CI = 0.94-1.70). This relationship was more pronounced in spontaneous births (RR = 1.58; 95% CI = 0.96-2.61) and emergency cesarean sections (RR = 1.76; 95% CI = 1.10-2.82). Significance persisted after adjusting for numerous potential confounders. CONCLUSION: Our results confirm an increased mortality risk for night-time births which could not be explained by other accessible risk factors. This suggests that the increased risk at night might be attributable to a reduced availability to provide appropriate medical care in delivery units at night. DISCUSSION: As mainly the presence of staff is decreased during the night, introduction of better designed shifts can be expected to reduce neonatal mortality. PMID- 14528417 TI - [Mortality and morbidity of preterm infants--a synopsis of the german neonatal census 2001]. AB - BACKGROUND: The German Neonatal Census provides epidemiological data in the field of perinatology particularly concerning the rates of preterm birth and the mortality and morbidity of preterm infants. MATERIALS: All but two inquiries in Germany in 2001 were analyzed, covering 87% of all live births. RESULTS: The transfer rate from obstetric departments to children's hospitals/neonatological wards was 14.6% of which 87.4% were singletons, 11.5% were twins and 1.1% were triplets or higher order births. 1.3% of all admitted patients were born before 32 weeks gestational age and the mortality rate was 9.1%. After 28 weeks gestational age the mortality rate was lower for twins than for singletons, after 32 weeks gestational age it was lower for triplets as well. In 17.4% of all patients born before 28 weeks gestational age a high grade intracranial hemorrhage (B III and IV) was found and in 6.2% a periventricular leukomalacia was seen. 15.8% of all surviving preterm infants before 28 weeks developed ROP 3 + >. CONCLUSIONS: A synopsis of all neonatal censuses is an appropriate tool to study morbidity and mortality in preterm infants. Comparing the results with published data of large cohorts make the presented results reasonable although documentation seems to need some improvement in several fields. PMID- 14528418 TI - [Maternal risks for newborn macrosomia, incidence of a shoulder dystocia and of damages of the plexus brachialis]. AB - PATIENTS RESPECTIVELY AND METHODS: In the HEPE 619 242 births have been analysed (1990 - 2000) to calculate the incidence of a birthweight between 4000 and 4499 g and of a weight > or = 4500 g in relationship to maternal obesity, high maternal weight gain and of a duration of pregnancy more than 298 days. RESULTS: The risk of a macrosomia > or = 4500 g is in cases of obesity 3.4 times higher, in cases of obesity and prolongation of pregnancy 6.6 times higher and in the presence of all 3 risk factors 10 times higher. Data of the Frauenklinik Wiesbaden (HSK) (n = 6075 births) complete the results, because a correlation between macrosomia, shoulder dystocia and a damage to the plexus brachialis has been found. The incidence of a damage to the plexus brachialis is in case of a shoulder dystocia and a birth weight of < or = 4000 g 6.3%, at a birth weight between 4000-4499 g 25% and in newborns with a weight > or = 4500 g 40%. CONCLUSIONS: Because of the low sensitivity (60%) of the ultrasonic weight measurement the 3 maternal risk factors--if they exist--of a shoulder dystocia and of a damage of the plexus brachialis should be discussed with the pregnant woman to help her about the decision of an alternative cesarean section. Still one third of the newborns weigh more than 4000 g if all 3 maternal risk factors exist. PMID- 14528419 TI - [Hepatitis and pregnancy--a short review]. AB - Viral hepatitis is one of the most serious infections in pregnant women. There are now at least five hepatitis viruses. This educational review describes the various types of hepatitis, clinical manifestations, their impact on mother and fetus, prevention and treatment options, risks of perinatal transmission and differential diagnosis. Several open questions regarding vaccination as well as perinatal transmission have been addressed. PMID- 14528420 TI - Advances in adult living donor liver transplantation. PMID- 14528421 TI - Selection of donors for living donor liver transplantation. PMID- 14528422 TI - Psychosocial assessment of adult living liver donors. PMID- 14528423 TI - Financial impact of adult living donation. PMID- 14528424 TI - Evaluation of the donor liver for living donor liver transplantation. PMID- 14528425 TI - Surgical techniques used in adult living donor liver transplantation. PMID- 14528426 TI - Hepatic regeneration in living donor liver transplantation. PMID- 14528427 TI - Donor and recipient outcomes after adult living donor liver transplantation. PMID- 14528428 TI - Early and late complications in the right-lobe adult living donor. PMID- 14528429 TI - Early and late complications in the recipient of an adult living donor liver. PMID- 14528430 TI - The impact of the model for end-stage liver disease on recipient selection for adult living liver donation. PMID- 14528431 TI - Role of adult living liver donation in patients with hepatocellular cancer. PMID- 14528432 TI - Role of adult living donor liver transplantation in patients with hepatitis C. PMID- 14528433 TI - Regulation and oversight of adult living donor liver transplantation. PMID- 14528434 TI - Future of adult living donor liver transplantation. PMID- 14528436 TI - Sonographic evaluation of the anterior liver surface in chronic liver diseases using a 7.5-MHz annular-array transducer: correlation with laparoscopic and histopathologic findings. AB - PURPOSE: We studied the sonographic appearance of the anterior liver surface using an ultrasound scanner equipped with a 7.5-MHz annular-array transducer to determine the accuracy of this imaging modality in monitoring the course of chronic liver diseases. METHODS: We prospectively evaluated patterns of the liver surface in the sonograms of 77 consecutive patients with chronic liver diseases who had undergone sonographic examination with a 7.5-MHz annular-array transducer and a 3.75-MHz convex-array transducer over a 2-year period and compared these findings with those of laparoscopy (using previously described categories) and histopathology. RESULTS: Histopathologically confirmed disease prevalences for inactive chronic hepatitis, active chronic hepatitis, liver cirrhosis, and others were 10% (8/77), 56% (43/77), 29% (22/77), and 5% (4/77), respectively. The sonographic appearance of the liver surface with the 3.75-MHz transducer was classified as either a regular or an irregular pattern. The regular pattern corresponded to 69% (34/50) of the cases in laparoscopic category 200 or 300 and the irregular pattern with 85% (23/27) of the cases in category 400 or 500. The sonographic appearance of the liver surface with the 7.5-MHz transducer was classified as regular, unevenly irregular, diffusely irregular, or nodular. These 4 patterns detected 75% (24/32), 78% (14/18), 52% (12/23), and 75% (3/4) of the cases of laparoscopic categories 200, 300, 400, and 500, respectively. In a comparison of the sonographic patterns of the liver surface with the differential histopathologic findings, the regular sonographic pattern corresponded to 88% (7/8) of the cases of inactive chronic hepatitis, the unevenly irregular pattern with 35% (15/43) of the cases of active chronic hepatitis, and the diffusely irregular and nodular patterns (considered as 1 group) with 68% (15/22) of the cases of liver cirrhosis. CONCLUSIONS: Our results suggest that sonographic evaluation of the liver surface with a 7.5-MHz annular-array transducer using this classification provides detailed information on the evolution of chronic liver diseases that correlates with the laparoscopic and histopathologic findings and thus is a useful noninvasive method for monitoring the disease course to cirrhosis. PMID- 14528437 TI - Transvaginal sonographic appearance of thrombosed uterine arteries after uterine artery embolization: the "white snake" sign. AB - PURPOSE: The aim of this prospective study was to describe the appearance of thrombosed uterine arteries on transvaginal sonography performed after uterine artery embolization (UAE) and to assess the prognostic value of the "white snake" sign with regard to symptomatic outcome at 12 months. METHODS: Patients who underwent UAE from January 1, 1999, to July 31, 2000, for the treatment of symptomatic leiomyomas were included in the study. Transvaginal sonography was performed before and at 3, 6, and 12 months after UAE. Patients graded the severity of their symptoms on a scale from 1 to 5, with 1 being the least and 5 the most severe, before and at 12 months after the procedure. The Wilcoxon rank sum test was used to determine correlations between severity of symptoms and presence of the white snake sign; a p value of less than 0.05 was considered significant. RESULTS: During the study period, UAE was performed in 19 patients with a mean age of 41 years (range, 32-48 years). UAE was technically successful in all patients. Eighteen patients (95%) reported symptomatic improvement at 12 months: 8 patients (42%) by 4 severity-scale points, 5(26%) by 3 points, and 5 (26%) by 2 points. The 1 patient who did not experience improvement had undergone a hysterectomy at 4 months after the UAE. At the 3-month follow-up, transvaginal sonography demonstrated a tortuous echogenic structure in the adnexa (the white snake sign) in all patients; the finding was still apparent in 10 patients at 6 months but in only 2 patients at 12 months. A direct correlation was found between persistence of the white snake sign and the degree of symptomatic improvement at 6 months (p=0.04) but not at 12 months (p=0.08). CONCLUSIONS: After UAE, a thrombosed uterine artery appears on transvaginal sonography as an echogenic tortuous structure in the adnexa. Persistence of this white snake sign at 6 months after UAE may suggest a more favorable symptomatic outcome. PMID- 14528438 TI - Diagnosis of arterial occlusive disease of the upper extremities: comparison of color duplex sonography and angiography. AB - PURPOSE: The aim of this study was to assess the abilities of color duplex sonography (CDUS) to detect and characterize arterial occlusive disease of the upper extremities. METHODS: We prospectively compared the results of CDUS with those of intra-arterial digital subtraction angiography, which were considered definitive, in patients with symptomatic arterial occlusive disease of the upper extremities. In each extremity, we visualized 9 arterial segments, which were each evaluated for stenoses and occlusions. Each segment was categorized on each imaging modality as not significantly narrowed (narrowed by <50%), significantly narrowed (narrowed by > or =50% but <100%), or occluded (100%). RESULTS: We examined a total of 578 segments in 57 patients (34 men and 23 women) with a mean age of 50 years (range, 20-74 years). CDUS had a sensitivity, a specificity, a positive predictive value, a negative predictive value, and an accuracy of 98%, 99%, 97%, 99.5%, and 99%, respectively, for detecting occluded lesions and 79%, 100%, 100%, 99%, and 99%, respectively, for detecting hemodynamically significantly stenotic lesions. Notably, the sensitivity of CDUS for diagnosing significantly stenotic lesions (79%) was lower than that for diagnosing occlusive disease (98%). CONCLUSIONS: With high sensitivity and accuracy rates, CDUS is a reliable screening method for detecting arterial occlusive disease of the upper extremities. This modality efficiently provides anatomic and hemodynamic data that are useful in cases of such disease. PMID- 14528439 TI - Real-time three-dimensional echocardiography versus two-dimensional echocardiography in the diagnosis of left ventricular apical thrombi: preliminary findings. AB - PURPOSE: Real-time 3-dimensional transthoracic echocardiography (RT-3D-TE) with real-time volume rendering (RTVR) offers multiple simultaneous views and spatial definition of intracardiac structures superior to that attainable by 2 dimensional transthoracic echocardiography (2D-TE). We hypothesized that RT-3D-TE would therefore improve identification of left ventricular apical thrombi (LVT). METHODS: Patients were referred to our echocardiography laboratory over an 8 month period. Those diagnosed with a "suspicious" or "definite" LVT on the basis of 2D-TE images underwent RT-3D-TE on the same day. All 2D-TE, RT-3D-TE, and RTVR images were reviewed by 2 independent observers. RT-3D-TE findings were considered positive for LVT if LVT was visualized in both B-scan (apical orthogonal) and C-scan (short axis, with and without tilting angle) planes and on RTVR images, nondiagnostic (or suspicious) for LVT if it was not visualized in all planes, and negative for LVT if it was not visualized in any plane. RESULTS: Thirty patients (19 men and 11 women) with a mean age (+/- standard deviation) of 52 +/- 13 years were enrolled. The interobserver agreement coefficient was 63% for 2D-TE interpretations of LVT and 93% for RT-3D-TE interpretations of LVT (p<0.05). The final interpretations by RT-3D-TE with RTVR were positive for LVT in 16 patients (53%), suspicious for LVT in 4 patients (13%), and negative for LVT in 10 patients (30%). CONCLUSIONS: RT-3D-TE with RTVR offers dramatically clearer echocardiographic findings than does 2D-TE. In particular, RT-3D-TE is able to provide a clear diagnosis of LVT/non-LVT when 2D-TE images are merely suggestive of the disorder. Therefore, RT-3D-TE with RTVR, which is a clinically feasible alternative to 2-dimensional echocardiography, has great potential to positively affect the diagnosis, follow-up, and care of patients with suspected LVT. PMID- 14528440 TI - Echogenic fetal lung masses: comparison of prenatal sonographic and postnatal CT findings. AB - PURPOSE: The aim of this retrospective study was to compare changes in size and appearance of fetal lung masses detected on prenatal sonography with those on postnatal CT. PATIENTS AND METHODS: Sixteen fetuses with echogenic lung masses had undergone serial sonography in utero as well as postnatal CT scanning. Changes in size of the masses were determined by comparing initial and follow-up sonograms. The appearance of the lesions on postnatal CT scans was classified as consolidation, cavity, ground-glass opacity, or atelectasis. All sonographic and CT findings were analyzed by 2 experienced radiologists who were unaware of previous imaging results. RESULTS: The initial mean size of the fetal lung masses on prenatal sonography was 31 (anteroposterior dimension)x25(width)x36(craniocaudal length) mm. Follow-up serial sonography revealed complete regression of 10 masses, no change in 4, partial regression of 1, and increased size in 1. On postnatal chest CT, the mean mass size was 32x22x28 mm; 6 lesions showed no size change from that on the initial sonograms, 6 showed partial regression, and 4 showed an increase in size. Of the 10 cases with apparently complete regression on prenatal sonography, all showed positive findings on postnatal CT. CONCLUSIONS: The discrepancy between prenatal sonographic and postnatal CT findings supports the assumption that regression of lung masses during serial prenatal sonographic follow-up does not always indicate their complete regression. PMID- 14528441 TI - Secretory carcinoma of the breast. AB - Secretory carcinoma of the breast is a rare but histologically distinct variant of invasive ductal carcinoma that has an indolent growth pattern and a more favorable prognosis than that of typical ductal carcinoma. We present the imaging and histopathologic findings of a case of this tumor that initially manifested as a palpable mass in the left breast of a 31-year-old woman. It appeared sonographically as a round, well-circumscribed but microlobulated hypoechoic solid mass. Secretory carcinoma of the breast may resemble other well circumscribed breast carcinomas as well as some benign masses, including fibroadenomas. Breast imagers should be aware of this rare tumor's nonspecific imaging features and clinical implications when making a differential diagnosis of solid breast masses. PMID- 14528442 TI - Cavernous hemangioma of the breast: mammographic and sonographic findings and follow-up in a patient receiving hormone-replacement therapy. AB - We report the case of a 78-year-old woman who had been receiving hormone replacement therapy (HRT) for 6 years and had a 4-month history of a painless nodule in the 9 o'clock position in her right breast. Mammography performed 4 years previously had shown a 4-mm bilobed, ovoid, well-defined nodule in that location; mammography performed 1 year previously had shown that the nodule had increased to 6 mm. We performed mammographic and sonographic examinations, which revealed a 10-mm ovoid nodule in the same 9 o'clock position in the right breast. The imaging findings appeared to indicate benignity, but because of the increasing size of the nodule, we undertook an ultrasound-guided large-core needle biopsy. The histopathologic diagnosis was typical cavernous hemangioma. It was not excised, but HRT was discontinued. Follow-up mammography and sonography 8 months later showed that the nodule had decreased to 6 mm. We believe that the HRT played a contributory role in the increasing size of this patient's cavernous hemangioma. The use of ultrasound-guided large-core needle biopsy is reliable enough to ascertain the benignity of such masses and can thus avoid, if it is clinically appropriate, the need for their surgical removal. PMID- 14528443 TI - Mirror-image artifact mimicking epidural hematoma: usefulness of power Doppler sonography. AB - During sonographic examination, a mirror-like interface can scatter the ultrasound beam, producing mirror-image artifacts. We describe a case of bilateral parietal cephalhematoma in the parietal region of a 2-day-old neonate that mimicked bilateral epidural hematoma on routine sonographic examination. Forceps had been used during delivery, and the neonate had been experiencing episodes of apnea since birth. Gray-scale coronal sonograms showed anechoic structures resembling epidural hematomas in the left and right parietal regions. However, color and power Doppler sonography showed normal vascular findings, thus permitting identification of the mirror-image artifact. PMID- 14528444 TI - Treatment of a radial artery pseudoaneurysm with ultrasound-guided percutaneous thrombin injection in a patient with Behcet's syndrome. AB - A 57-year-old man with Behcet's syndrome and recurrent deep vein thrombosis of the lower limbs presented with a painful, pulsating mass on the volar aspect of the radial edge of his left wrist. One month before this visit, he had had venous blood drawn from the same site. Using color Doppler sonography, we diagnosed an iatrogenic pseudoaneurysm of the left radial artery, which was then treated with an ultrasound-guided percutaneous injection of thrombin. A follow-up examination 6 months after the treatment revealed complete resolution of the pseudoaneurysm. To our knowledge, this is the first case report to demonstrate the use of this technique for thrombosis of a pseudoaneurysm in a patient with Behcet's syndrome. We believe that the safety, efficiency, speed, and minimal invasiveness of this procedure make it feasible for use as a treatment for peripheral pseudoaneurysms in such patients. PMID- 14528445 TI - Isolated cardiomegaly in the second trimester as an early sign of fetal hydrops due to intracranial arteriovenous malformation. AB - We report the case of a 27-year-old pregnant woman in whom isolated mild fetal cardiomegaly, diagnosed prenatally on sonographic examination at 22 weeks' menstrual age, was the first sign of development of an arteriovenous malformation of the vein of Galen. The arteriovenous malformation was visualized on sonographic examination at 29 weeks' menstrual age; prenatal MRI at 32 weeks confirmed the diagnosis. At 34 weeks' menstrual age, repeat sonographic examination demonstrated polyhydramnios, cardiomegaly, and generalized hydrops with ascites and pleural effusion. After vaginal delivery, the male neonate died of respiratory distress. We present the findings of prenatal gray-scale, color Doppler, and power Doppler sonography; MRI; and postmortem examination. PMID- 14528446 TI - Parcellation of cortical afferents to three distinct sectors in the parahippocampal gyrus of the rhesus monkey: an anatomical and neurophysiological study. AB - The posterior parahippocampal gyrus (PHG) of the rhesus monkey (Macaca mulatta) is comprised of three distinct cortical areas based on cytoarchitecture, connectivity, and neurophysiological response properties. Fluorescent retrograde tracers placed in each PHG area demonstrated unique patterns of cortical afferent input to areas TH, TL, and TF. Area TF receives inputs from the multimodal cortices of the superior temporal sulcus including areas PGa, TPO, and MST, from the visuospatial parietal area PG-Opt, and from visual areas V3A and dorsal V4. Area TL receives afferents from the inferotemporal region including visual areas TE1 and TE2 as well as from areas TEa, IPa, and FST in the lower bank and depth of the superior temporal sulcus. In contrast, the input to area TH is from the rostral part of superior temporal gyrus, including the auditory association areas TS1-3, and from the middle sector of area TPO in the superior temporal sulcus. Frontal and cingulate areas also project to the PHG in largely differential patterns. To further investigate this a correlative electrophysiological study of the three PHG areas resulted in a confirmation of these differential cortical inputs such that visually responsive neurons were found in areas TF and TL, auditory responsive neurons or bimodal auditory/visual-responsive neurons in area TH, and somatosensory-responsive neurons at the TF/TL border. Since each PHG area also receives differential hippocampal input, these data suggest that the processing of unimodal or multimodal information may be related to memory processing functions that are largely segregated within areas TH, TL, and TF. PMID- 14528447 TI - Organization and development of corticocortical associative neurons expressing the orphan nuclear receptor Nurr1. AB - The developmental mechanism that contributes to the highly organized axonal connections within the cerebral cortex is not well understood. This is partly due to the lack of molecular markers specifically expressed in corticocortical associative neurons during the period of circuit formation. We have shown previously that latexin, a carboxypeptidase A inhibitor, is expressed in intrahemispheric corticocortical neurons from the second postnatal week in the rat (Arimatsu et al. [1999] Cereb. Cortex 9:569-576). In the present study, we first demonstrate in the adult rat that the orphan nuclear receptor Nurr1 is coexpressed in latexin-expressing neurons located in layer V, sublayer VIa, and the white matter of the lateral sector of the neocortex, and also in latexin negative early born neurons in sublayer VIb of the entire neocortex. Virtually all Nurr1-expressing neurons exhibit immunoreactivity for phosphate-activated glutaminase but not for gamma-aminobutyric acid, suggesting that they are glutamatergic-excitatory neurons. By combining Nurr1 immunohistochemistry and 5 bromo-2'-deoxyuridine-birthdating, we then show that Nurr1 is expressed in (early born) subplate neurons and (later born) presumptive latexin-expressing neurons from embryonic day 18 onward. Finally, by combination of Nurr1 immunohistochemistry and retrograde tracing, we show that Nurr1-expressing neurons, including those in sublayer VIb, contribute predominantly to long-range intrahemispheric corticocortical projections. These results raise the possibility that Nurr1 plays a role in the establishment and maintenance of normal corticocortical circuitry and function. PMID- 14528448 TI - Development of the larval nervous system of the gastropod Ilyanassa obsoleta. AB - Gastropods have been well studied in terms of early cell cleavage patterns and the neural basis of adult behaviors; however, much less is known about neural development in this taxon. Here we reveal a relatively sophisticated larval nervous system in a well-studied gastropod, Ilyanassa obsoleta. The present study employed immunocytochemical and histofluorescent techniques combined with confocal microscopy to examine the development of cells containing monoamines (serotonin and catecholamine), neuropeptides (FMRFamide and leu-enkephalin related peptides), and a substance(s) reactive to antibodies raised against dopamine beta-hydroxylase. Neurons were first observed in the apical organ and posterior regions during the embryonic trochophore stage. During later embryonic development neurons appeared in peripheral regions such as the foot, velum, and mantle and in the developing ganglia destined to become the adult central nervous system. In subsequent free-swimming veliger stages the larval nervous system became increasingly elaborate and by late larval stages there existed approximately 26-28 apical cells, 80-100 neurons in the central ganglia, and 200 300 peripherally located neurons. During metamorphosis some populations of neurons in the apical organ and in the periphery disappeared, while others were incorporated into the juvenile nervous system. Comparisons of neural elements in other molluscan larvae reveal several similarities such as comparable arrangements of cells in the apical organ and patterns of peripheral cells. This investigation reveals the most extensive larval nervous system described in any mollusc to date and information from this study will be useful for future experimental studies determining the role of larval neurons and investigations of the cellular and molecular mechanisms governing neural development in this taxon. PMID- 14528449 TI - Expression of tryptophan 5-hydroxylase gene during sea urchin neurogenesis and role of serotonergic nervous system in larval behavior. AB - Tryptophan 5-hydroxylase (TPH) is the rate-limiting enzyme in the biosynthesis of serotonin. cDNA cloning of TPH was carried out, and the occurrence of spatiotemporal transcription of TPH message was examined in larvae of the sea urchin, Hemicentrotus pulcherrimus (HpTPH), with in situ hybridization by using the tyramide signal amplification (TSA) technique and Northern hybridization. Based on deduced amino acids sequence of HpTPH, phylogenetically sea urchin locates at the closest position to vertebrates among invertebrates, and HpTPH had common conserved sequences in a catalytic domain. Initiation of HpTPH transcription occurred at the late gastrula stage exclusively in serotonin cells of apical ganglion (SAG) that was composed of a cluster of HpTPH-positive cells and the negative cells in between. In situ hybridization showed that the mRNA expression pattern was similar to the immunohistochemical localization of serotonin cells reported before (Bisgrove and Burke [1986] Dev. Growth Differ. 28:557-569; Yaguchi et al. [2000] Dev. Growth Differ. 42:479-488). p Chlorophenylalanine (CPA), an irreversible inhibitor of TPH activity, considerably decreased serotonin content in the serotonin cells, whereas the HpTPH expression pattern and timing, and the extension of neurofibers from SAG cells were apparently unaffected, suggesting CPA exclusively perturbed synthesis of serotonin but not nervous system organization. CPA-treated larvae did not swim, despite the occurrence of ciliary beating in culture chamber, suggesting that proper serotonin synthesis is necessary for normal swimming of the larvae. PMID- 14528451 TI - Morphological analysis of disabled-1-immunoreactive amacrine cells in the guinea pig retina. AB - Disabled-1 (Dab1) is an adapter molecule in a signaling pathway, stimulated by reelin, that controls cell positioning in the developing brain. It localizes to selected neurons in the nervous system, including the retina, and Dab1-like immunoreactivity is present in AII amacrine cells in the mouse retina. This study was conducted to characterize Dab1-labeled cells in the guinea pig retina in detail using immunocytochemistry, quantitative analysis, and electron microscopy. Dab1 immunoreactivity is present in a class of amacrine cell bodies located in the inner nuclear layer adjacent to the inner plexiform layer (IPL). These cells give rise to processes that ramify the entire depth of the IPL. Double-labeling experiments demonstrated that these amacrine cells make contacts with the axon terminals of rod bipolar cells and that their processes make contacts with each other via connexin 36 in sublamina b of the IPL. In addition, all Dab1-labeled amacrine cells showed glycine transporter 1 immunoreactivity, indicating that they are glycinergic. The density of Dab1-labeled AII amacrine cells decreased from about 3,750 cells/mm(2) in the central retina to 1,725 cells/mm(2) in the peripheral retina. Dab1-labeled amacrine cells receive synaptic inputs from the axon terminals of rod bipolar cells in stratum 5 of the IPL. From these morphological features, Dab1-labeled cells of the guinea pig retina resemble the AII amacrine cells described in other mammalian species. Thus, the rod pathway of the guinea pig retina follows the general mammalian scheme and Dab1 antisera can be used to identify AII amacrine cells in the mammalian retina. PMID- 14528452 TI - Estrogen-receptor-beta distribution in the human hypothalamus: similarities and differences with ER alpha distribution. AB - This study reports the first systematic rostrocaudal distribution of estrogen receptor beta immunoreactivity (ER beta-ir) in the human hypothalamus and adjacent areas in five males and five females between 20-39 years of age and compares its distribution to previously reported ER alpha in the same patients. ER beta-ir was generally observed more frequently in the cytoplasm than in the nucleus and appeared to be stronger in women. Basket-like fiber stainings, suggestive for ER beta-ir in synaptic terminals, were additionally observed in various areas. Men showed more robust nuclear ER beta-ir than women in the medial part of the bed nucleus of the stria terminalis, paraventricular and paratenial nucleus of the thalamus, while less intense, but more nuclear, ER beta-ir appeared to be present in, e.g., the BSTc, sexually dimorphic nucleus of the medial preoptic area, diagonal band of Broca and ventromedial nucleus. Women revealed more nuclear ER beta-ir than men of a low to intermediate level, e.g., in the suprachiasmatic, supraoptic, paraventricular, infundibular, and medial mamillary nucleus. These data indicate potential sex differences in ER beta expression. ER beta-ir expression patterns in subjects with abnormal hormone levels suggests that there may be sex differences in ER beta-ir that are "activational" rather than "organizational" in nature. Similarities, differences, potential functional, and clinical implications of the observed ER alpha and ER beta distributions are discussed in relation to reproduction, autonomic-function, mood, cognition, and neuroprotection in health and disease. PMID- 14528453 TI - Collateralization of climbing and mossy fibers projecting to the nodulus and flocculus of the rat cerebellum. AB - Collateralization of mossy and climbing fibers was investigated using cortical injections of cholera toxin b-subunit in the rat vestibulocerebellum. Injections were characterized by their retrograde labeling within the inferior olive. Collateral labeling was plotted using color-coded density profiles of the whole cerebellar cortex. Injections in the medial part of the nodulus resulted in olivary labeling that was restricted to the rostral part of the dorsal cap. Climbing fiber collaterals were found in medial and lateral nodular zones as well as in the ventral paraflocculus and adjacent flocculus. Injections in the intermediate part of the nodulus resulted in olivary labeling of the beta subnucleus but could also involve the ventrolateral outgrowth. In the latter case, climbing fiber collaterals were found in the two floccular zones and in a small region in the lateral-most part of crus I. All nodular injections showed a bilaterally symmetric distribution of collateral mossy fiber rosettes that was mostly confined to the vestibulocerebellum and originated predominantly from the vestibular nuclei. Injections in the flocculus labeled the caudal part of the dorsal cap and/or the ventrolateral outgrowth. Mossy fiber rosettes were observed throughout the vestibulocerebellum but also included other regions of the cerebellar cortex in a bilaterally symmetric pattern corresponding with a more widespread precerebellar origin. Climbing fibers originating in the rostral dorsal cap, labeled from an injection in the ventral paraflocculus, collateralize to a medial and lateral zone in the nodulus. Climbing fiber collaterals were usually accompanied by subjacent labeling of mossy fiber rosettes. These results demonstrate that some nodular and floccular zones are related and, at least partially, share a common input. PMID- 14528450 TI - Intercellular interactions in the mammalian olfactory nerve. AB - The small, unmyelinated axons of olfactory sensory neurons project to the olfactory bulb in densely packed fascicles, an arrangement conducive to axo axonal interactions. We recently demonstrated ephaptic interactions between these axons in the olfactory nerve layer, the layer of the olfactory bulb in which the axon fascicles interweave and rearrange extensively. In the present study, we hypothesized that the axons, which express connexins, may have another mode of communication: gap junctions. Previous transmission electron microscopy (TEM) studies have failed to demonstrate such junctions. However, the definitive method for detecting gap junctions, freeze fracture, has not been used to examine the interaxonal connections of the olfactory nerve layer. Here, we apply a combined approach of TEM and freeze fracture to determine if gap junctions are present between the olfactory axons. Gap junctions involving olfactory axons were not found. However, by freeze fracture, P faces of both the axons and ensheathing cells (glia that surround the axon fascicles) contained distinctive linear arrays of particles, aligned along the small columns of extracellular space. In axons, few intramembranous particles were present outside of these arrays. Multi-helix proteins, including ion channels and connexin hemichannels, have been shown to be visible as particles by freeze fracture. This suggests that the proteins important for signal transmission are confined to the linear arrays. Such an arrangement would facilitate ephaptic transmission, calcium waves, current oscillations, and paracrine communication and may be important for olfactory neural code processing. PMID- 14528454 TI - Temperature elevation after vertebroplasty with polymethyl-methacrylate in the goat spine. AB - Although the general complication rate for vertebroplasty is low, neural compression and thermal damage have been related to the use of polymethyl methacrylate cement. This study focuses on the risk of thermal tissue damage after vertebroplasty. In this study, cavities were created by a transpedicular approach in the vertebral bodies (L3, L4, and L5) of four milk goats, and polymethyl-methacrylate cement was injected. In three locations (the bone-cement interface, the epidural space, and the disc space) the temperature was measured in regular intervals after injection of the cement. The mean injected volume was 0.8 ml, which accounted for a 22% volume fraction. The mean peak temperature of the bone-cement interface was 44.6 degrees C, while the maximum temperature at the epidural space and disc space was 37.0 and 37.5 degrees C, respectively. The local temperature measured after in vivo vertebroplasty did not reach values that are known to cause tissue necrosis. PMID- 14528455 TI - Three-dimensional hyaluronic acid grafts promote healing and reduce scar formation in skin incision wounds. AB - Hyaluronic acid (HA) has been found to play important roles in tissue regeneration and wound-healing processes. Fetal tissue with a high concentration of HA heals rapidly without scarring. The present study employed HA formed into three-dimensional strands with or without keratinocytes to treat full-thickness skin incision wounds in rats. Wound closure rates of HA strand grafts both with and without keratinocytes were substantially enhanced. The closure times of both HA grafts were less than 1 day (average 16 h), about 1/7 that of the contralateral control incisions (114 h, p <.01). Average wound areas after 10 days were HA-only graft: 0.151 mm2 +/- 0.035; HA + cell grafts: 0.143 mm2 +/- 0.036 and controls: 14.434 mm2 +/- 1.175, experimental areas were 1% of the controls (p < 0.01). Transforming growth factor (TGF) beta1 measured by immunostaining was remarkably reduced in HA-treated wounds compared to the controls. In conclusion, HA grafts appeared to produce a fetal-like environment with reduced TGF-beta1, which is known to be elevated in incipient scars. The HA strands with or without cultured cells may potentially improve clinical wound healing as well as reduce scar formation. PMID- 14528456 TI - A new model of implant-related osteomyelitis in rats. AB - Infection related to osteosynthesis often has dramatic consequences for the patient. Prolonged hospitalization with systemic antibiotic therapy, several revision procedures, possible amputation, and even death may occur. To investigate the pathology of infection in orthopedic surgery, a new rat model of implant related osteomyelitis was developed. Three different concentrations (10(6), 10(3), and 10(2) colony-forming units (CFU)/10 microl) of Staphylococcus aureus were inoculated into the tibial medullary cavity with simultaneous insertion of a titanium Kirschner wire. Controls received phosphate-buffered saline (PBS). Each group consisted of 10 animals. Animals were followed for 4 weeks until sacrifice. X-rays of the tibiae were taken weekly, blood counts were analyzed, and body temperature and weight were determined. After sacrifice, infection was evaluated by histological and microbiological investigations. All animals inoculated with Staph. aureus in either concentration developed microbiological, histological, and radiological signs of osteomyelitis in correlation to the amount of inoculated bacteria. X-rays clearly revealed osseous destruction after 14 days with progression of osteomyelitis during the following weeks. CFU/g bone and bone weight after sacrifice showed dependence on the amount of inoculated CFU. The histological results confirmed the radiological findings. No significant changes in blood counts, body weight, and body temperature between the groups could be observed. The results demonstrate that it is possible to develop a model of implant-related osteomyelitis in rats with dependence on the amount of inoculated bacteria. No other promoters of infection besides intramedullary insertion of titanium Kirschner wires were used in this model. PMID- 14528457 TI - An animal evaluation of a paste of chitosan glutamate and hydroxyapatite as a synthetic bone graft material. AB - The objective of this study was to develop a synthetic bone graft in a paste form. Reported here are the results of the evaluation of a paste of chitosan glutamate (Protosan) and hydroxyapatite (referred to as a paste) used in a critical size defect model in rats. Eight-millimeter--diameter cranial defects were made in rat calvaria following a protocol approved by the animal review committee. Five groups were studied: (1) empty control, (2) defect filled with paste only, (3) defect filled with the paste containing bone-marrow aspirate, (4) defect filled with paste containing BMP-2, and (5) defect filled with paste containing osteoblasts cultured from bone-marrow aspirate. The sacrifice intervals were 9 and 18 weeks. Calvaria containing the defect were harvested, and the bone mineral density (BMD) was determined by dual energy X-ray absorptiometry. Push-out strength measurements were also performed. The BMD values of empty control were significantly lower than those of other groups at both 9 and 18 weeks. The mechanical properties, that is, push-out strengths and area under the push-out load and displacement were not significantly different between the samples. Histological examination of Goldner-trichromestained undecalcified sections showed the presence of mineralized bone spicules in the defect areas that were more prominent in those filled with paste and osteoblasts cultured from bone-marrow aspirate. Hence, this study demonstrated that the paste of chitosan glutamate and hydroxyapatite-containing osteoblasts cultured from bone-marrow aspirate would be an effective material to repair bone defects. PMID- 14528458 TI - Paraffin spheres as porogen to fabricate poly(L-lactic acid) scaffolds with improved cytocompatibility for cartilage tissue engineering. AB - Three-dimensional poly(L-lactic acid) (PLLA) scaffolds with high porosity and pore size ranging from 150 to 700 microm were conveniently prepared with paraffin spheres used as porogen. PLLA/1,4-dioxane solution containing a given amount of paraffin spheres was frozen at -25 degrees C to obtain a solidified mixture, followed with freeze drying and subsequent leaching with hexane to remove the 1,4 dioxane and paraffin spheres, respectively. The fabricated PLLA scaffolds were highly porous with evenly distributed and interconnected pores. The microstructures of the PLLA scaffolds as a function of paraffin-sphere size, paraffin-sphere dosage, and PLLA concentration were characterized by confocal laser scanning microscopy (CLSM) and scanning-electronic microscopy (SEM). To improve the cytocompatibility of the bioinert material, a hybrid PLLA scaffold containing Type I collagen was prepared by pressing the collagen solution into the scaffold under reduced pressure. The amounts of the collagen introduced in the scaffolds were detected by ninhydrin method. The distribution of the collagen in the scaffolds was studied with CLSM. Finally, in vitro cell culture was performed by injecting a chondrocyte suspension into the scaffolds. The results showed that the chondrocytes were more evenly distributed and more spread out in the collagen-modified PLLA scaffolds than in the unmodified ones. PMID- 14528459 TI - Composite surgical sutures with bioactive glass coating. AB - A processing method was developed to coat polyglactin 910 (Vicryl) sutures with bioactive glass powder (45S5 Bioglass). High reproducibility and homogeneity of the coating in terms of microstructure and thickness along the suture length were achieved. Bioglass-coated sutures exhibited a high level of chemical reactivity in simulated body fluid (SBF), indicating their bioactive behavior. This was evident by the prompt formation of hydroxyapatite (HA) crystals on the surface after only 7 days of immersion in SBF. These crystals grew to form a thick HA layer (15 microm thickness) after 3 weeks in SBF. The tensile strength of the sutures was tested before and after immersion in SBF in order to assess the effect of the bioactive glass coating on suture degradation. The tensile strength of composite sutures was lower than that of as-received Vicryl sutures, 385 and 467 MPa, respectively. However, after 28 days of immersion in SBF the residual tensile strengths of coated and uncoated sutures were similar (83 and 88 MPa, respectively), indicating no negative effect of the HA layer formation on the suture strength. The effect of bioactive glass coating on the polymer degradation is discussed. The developed bioactive sutures represent interesting materials for applications in wound healing, fabrication of fibrous three-dimensional scaffolds for tissue engineering, and reinforcement elements for calcium-phosphate temporary implants. PMID- 14528460 TI - Influence of dental biofilm on release of mercury from amalgam exposed to carbamide peroxide. AB - Tooth bleaching is a popular procedure in modern aesthetic dentistry. Bleaching agents may affect amalgam restorations by altering the release of mercury. The aim of this study was to explore the effect of biofilm-coated amalgam restorations on the release of mercury in the presence of carbamide peroxide. Samples of SDI and Valliant amalgams were submerged for either 14 days or 7 months in buffered KCl after which they were coated with saliva, bacteria, and polysaccharides. The samples were exposed to 10% carbamide peroxide (CP) for 24 h. The amount of mercury released was examined for 120 h. Results showed that most of mercury release occurred within the first 24 h, after which the release rate decreased sharply. After 120 h the release of mercury from the tested samples was minimal and similar to the control group. The presence of biofilm coating on the amalgam samples did not induce the release of mercury but tended to reduce mercury release into the surrounding environment. CP induces the release of mercury from amalgam samples. However, the presence of biofilm did not prevent large amounts of mercury release from amalgam coated with biofilms and exposed to CP. This study indicates that dental biofilm may retard the release of mercury from amalgam restorations. PMID- 14528461 TI - Phase evolution during the formation of stoichiometric hydroxyapatite at 37.4 degrees C. AB - The mechanism of formation of stoichiometric hydroxyapatite in aqueous solution by the acid-base reaction of the precursor calcium phosphates tetracalcium phosphate [Ca4(PO4)2O, TetCP] and anhydrous dicalcium phosphate (CaHPO4, DCPA) was investigated. Phase evolution during the formation of hydroxyapatite was studied, and could be correlated with the kinetics of reaction. Initial dissolution of precursors is followed by hydroxyapatite nucleation and growth. Consumption of DCPA precedes that of TetCP and results in the formation of nonstoichiometric hydroxyapatite. As a result of continued TetCP dissolution, the pH of the solution increases towards a pH at which hydroxyapatite and Ca(OH)2 co exist in equilibrium. Hydroxyapatite formation takes place in two stages: initially formed, nonstoichiometric hydroxyapatite subsequently reacts with TetCP and is converted to stoichiometric hydroxyapatite. PMID- 14528462 TI - Clinical and simulator wear study of alumina ceramic THR to 17 years and beyond. AB - Three THAs with cementless monolithic alumina ceramic sockets and cementless Co alloy stems were retrieved because of aseptic loosening after 17 and 24 years. At revision heads and cups were marked for orientation. Maps were drawn of wear patterns with the use of light microscopy and surveyed by SEM. In a simulator experiment 28-mm-diameter alumina heads and liners were used. The cups were mounted inverted in a hip simulator and run with calf serum as the lubricant. The hip loads were 2 kN maximum and a 1-Hz frequency for 20 million cycles. Wear severity was classified into five grades. In retrieved implants, SEM analysis showed that the main wear zones (MWZ) had Grade 4 wear. The peripheral wear zones (PWZ) showed grain pull-out regions (Grade 5 wear). These corresponded to neck socket impingement and head-acetabular cup separation. Gray was due to transferred CoCr particles from the stem. In the simulator study, the MWZ had only localized areas of grain pull out surrounded by polished surface regions (Grade 4 wear) at 20 million cycles; stripe wear was not seen. The alumina ceramic bearings proved excellent up to 22 years in simulator studies and clinical studies. However, microseparation kinematics would be necessary in the simulator to duplicate the more peripheral wear zones. PMID- 14528463 TI - Host protection against deliberate bacterial contamination of an extracellular matrix bioscaffold versus Dacron mesh in a dog model of orthopedic soft tissue repair. AB - The resistance of two biomaterials, one synthetic and one biologic in origin, to deliberate bacterial infection was compared in a dog model of orthopedic soft tissue reconstruction. Twenty-four adult female dogs were randomly divided into two equal groups and a 2.0-cm-round full-thickness defect was created on the lateral surface of the stifle joint, leaving only the synovium and skin intact. The defect was surgically repaired with either Dacron trade mark mesh or a porcine derived extracellular matrix (ECM) scaffold material. The repair site was inoculated with 1 x 10(8) Staphylococcus aureus at the time of surgery and the dogs were survived for 28 days. Results showed a chronic pyogranulomatous inflammatory response at the Dacron trade mark implant sites versus a constructive tissue-remodeling response without residual inflammation at the ECM implant site. Three dogs in the group receiving the Dacron trade mark mesh were treated with Keflex trade mark (500 mg bid x 7 days) for signs of septicemia. A quantitative bacterial count of the implant sites at the time of sacrifice showed 6.52 x 10(5) +/- 1.2 x 10(6) and 6.5 x 10(2) +/- 1.8 x 10(3) bacteria per gram of tissue for the Dacron trade mark and ECM scaffold sites, respectively (P <.03). The ECM implant material was more resistant than the synthetic implant material to persistent infection following deliberate bacterial contamination and the ECM scaffold supported constructive tissue remodeling. PMID- 14528464 TI - Osteointegration of biomimetic apatite coating applied onto dense and porous metal implants in femurs of goats. AB - Biomimetic calcium phosphate (Ca-P) coatings were applied onto dense titanium alloy (Ti6Al4V) and porous tantalum (Ta) cylinders by immersion into simulated body fluid at 37 degrees C and then at 50 degrees C for 24 h. As a result, a homogeneous bone-like carbonated apatitic (BCA) coating, 30 microm thick was deposited on the entire surface of the dense and porous implants. Noncoated and BCA-coated implants were press-fit implanted in the femoral diaphysis of 14 adult female goats. Bone contact was measured after implantation for 6, 12, and 24 weeks, and investigated by histology and backscattered electron microscopy (BSEM). After 6 weeks, bone contact of the BCA-coated Ti6Al4V implants was about 50%. After 12 and 24 weeks, bone contact was lower in comparison with the 6-week implantations at, respectively 24 and 39%. Regarding the BCA-coated porous Ta implants, bone contacts were 17, 30, and 18% after 6, 12, and 24 weeks, respectively. However, bone contact was always found significantly higher for BCA coated dense Ti6Al4V and porous Ta cylinders than the corresponding noncoated implants. The results of this study show that the BCA coating enhances the bone integration as compared to the noncoated implants. PMID- 14528466 TI - Ultrasound in assessment of fetal growth disorders: is there a role for subcutaneous measurements? PMID- 14528465 TI - Polydimethylsiloxane versus polytetrafluoroethylene for vocal fold medialization: histologic evaluation in a rabbit model. AB - The objective is to study the tissue reaction of the paralyzed vocal cord in response to the injection of particulate plastics in a rabbit model. Forty-five New Zealand rabbits with surgical vocal-fold paralysis were used in the study. Histologic reactions of the larynx and the regional lymph nodes were analyzed by a single blinded pathologist at 6 weeks and 6 months after a vocal-cord injection of Teflon or of silicone elastomer. Macroscopic studies of the liver, lungs, spleen, kidney, and brain were performed. The histological study showed a greater proportion of chronic granulomatous inflammation in animals injected with silicone than in those injected with Teflon. The immunohistochemical study showed a higher degree of phagocytosis of Teflon particles than of the silicone particles. The silicone group presented a more severe fibrous reaction than the Teflon group, but the difference was not significant. No migration particles were found. It is concluded that silicone, having a greater viscosity than Teflon because of the size of its particles, induces more fibrosis and a larger proportion of foreign giant cells in the host. Due to this histological reaction, silicone particles present greater anchorage and stability. PMID- 14528467 TI - Terminology of three-dimensional and four-dimensional ultrasound imaging of the fetal heart and other moving body parts. PMID- 14528468 TI - Venous Doppler ultrasound in 146 fetuses with congenital heart disease. AB - OBJECTIVE: To assess the influence of isolated congenital heart disease (CHD) on fetal venous Doppler blood flow velocity waveforms. METHODS: Doppler flow velocimetry was performed in the inferior vena cava and ductus venosus in 146 consecutive fetuses with antenatally diagnosed CHD. Gestational age ranged between 19 and 39 weeks. Fetuses with isolated CHD without non-immune hydrops fetalis (NIHF) (Group A, n = 89) were separated from seven fetuses showing isolated CHD with NIHF (Group B) and 50 cases complicated by chromosomal or other extracardiac malformations, intrauterine growth restriction or non-cardiogenic NIHF (Group C). The control group comprised 109 healthy fetuses of uncomplicated pregnancies. Individual peak velocity index for veins (PVIV) measurements were converted into their Z-scores (delta values) for statistical analysis. RESULTS: There was no statistical difference between fetuses with isolated CHD (Group A) and controls, for the delta PVIV of neither the ductus venosus nor the inferior vena cava. Statistical analysis revealed significant differences between non isolated CHD fetuses (Group C) and controls for both vessels. However, in a separate analysis of isolated right heart malformations compared with the remaining isolated heart malformations (Groups A and B), a significant difference was observed for the ductus venosus, but not the inferior vena cava. There was an overall survival of 62%. In Group A, 58% of fetuses survived despite increased PVIV and 22% of fetuses with normal venous Doppler had an adverse outcome. All fetuses with cardiogenic NIHF (Group B) died. CONCLUSION: Doppler studies of the ductus venosus and inferior vena cava in fetuses with isolated CHD do not present sufficient alterations to be a reliable marker for screening purposes for CHD in mid-second- and third-trimester fetuses. Furthermore, venous Doppler did not predict fetal outcome in cases of isolated CHD. Abnormal venous Doppler results were mainly attributable to myocardial dysfunction and also to severe right heart obstruction even in the absence of congestive heart failure. Therefore, venous Doppler studies are clinically helpful in indirectly monitoring cardiac function in fetuses with cardiac malformations. PMID- 14528469 TI - Retrograde aortic isthmus net blood flow and human fetal cardiac function in placental insufficiency. AB - OBJECTIVE: Retrograde aortic isthmus (AoI) net blood flow has been associated with diminished oxygen delivery to cerebral circulation. This study was designed to characterize the cardiac function in human fetuses with retrograde AoI net blood flow in pregnancies complicated by placental insufficiency. METHODS: The control group comprised 43 fetuses in uncomplicated pregnancies. Study groups consisted of fetuses with placental insufficiency, and either antegrade (Group 1; n = 18) or retrograde (Group 2; n = 11) AoI net blood flow. Volume blood flows (Q) of left (LVCO) and right (RVCO) ventricles, ductus arteriosus (Q(DA)), pulmonary arterial bed (Q(P)) and foramen ovale (Q(FO)) were calculated and their proportions (%) of combined cardiac output (CCO) were determined. Ventricular ejection forces were calculated. Blood velocity waveforms of the mitral (MV) and tricuspid (TV) valves were obtained. The proportion of left ventricular isovolumetric relaxation time (IRT%) of the cardiac cycle, and index of myocardial performance (IMP) were calculated. RESULTS: In Group 1, Q(DA)% was increased (P < 0.05) and Q(P)% decreased (P < 0.05) compared with the control group, and Q(FO)% was greater (P < 0.01) compared with the control group and Group 2. In Group 2, the distribution of CCO did not differ from that of the control group. Ventricular ejection forces were similar among the groups. In Group 2, the MV early filling/atrial contraction time-velocity integral ratio was greater (P < 0.05) compared with those of the control group and Group 1. In Groups 1 and 2, IRT% and IMP were increased (P < 0.001) compared with the control group. CONCLUSIONS: In placental insufficiency, fetuses with antegrade AoI net blood flow show a shift in RVCO from the pulmonary to the systemic circulation, and Q(FO) makes up the majority of LVCO. Fetuses with retrograde AoI net blood flow fail to demonstrate these changes, suggesting a relative drop in the oxygen content of the blood entering the left ventricle. PMID- 14528470 TI - The role of the three vessels and trachea view (3VT) in the diagnosis of congenital heart defects. AB - OBJECTIVE: To analyse the anomalies seen at the three vessels and trachea (3VT) view in fetuses with antenatally diagnosed congenital heart disease (CHD), specifically in defects involving the upper mediastinum, including the outflow tracts and/or arteries. METHODS: Seventy-four consecutive fetuses of singleton pregnancies with antenatally diagnosed structural heart defects were initially included in the study. We selected those in which there was an anomaly seen on the 3VT view and classified them into groups: abnormalities detected on the upper mediastinum (abnormal vessel size, alignment, arrangement and number), abnormal location of the transverse aortic arch in relation to the trachea, and abnormal functional behavior of the arterial vessels (reversed flow and turbulent flow) detected using color Doppler. The abnormalities detected were confirmed postnatally by a pediatric cardiologist, echocardiography and/or autopsy. RESULTS: Thirty-four of the 74 fetuses showed an abnormal 3VT view that could be classified into one of the groups. We classified the lesions according to the most evident abnormal finding during the examination. Gestational age ranged from 17 to 39 (mean, 30) weeks. Abnormal vessel size was found in 20 of the 74 fetuses, abnormal vessel alignment was found in two fetuses, and abnormal vessel arrangement in three fetuses. Two vessels instead of three were found in five fetuses and four vessels were found in two fetuses. Abnormal visualization of the trachea, between the pulmonary artery and the transverse arch, occurred in one fetus with isolated right aortic arch. We found reversed flow in the transverse aortic arch in three fetuses with aortic atresia and in two fetuses with severe coarctation of the aorta. Reversed flow in the pulmonary trunk was seen in a fetus with pulmonary atresia and double outlet right ventricle. Turbulent flow was found in two fetuses with outflow tract stenosis. CONCLUSION: Our study, descriptive in nature, demonstrates that the 3VT view is a reliable method with which to determine abnormalities in the upper mediastinum. It proved to be efficient in identifying an important group of critical heart defects involving the outflow tracts and the aortic arch. It also provided important clues to the diagnosis of minor anomalies considered as markers of other CHD. We believe that use of the 3VT view should become routine in screening for CHD. PMID- 14528471 TI - Fetal loss following ultrasound diagnosis of a live fetus at 6-10 weeks of gestation. AB - OBJECTIVE: To examine prospectively the value of demographic characteristics and ultrasound findings in the prediction of subsequent fetal loss in pregnancies with live fetuses at 6-10 weeks of gestation. METHODS: Transvaginal ultrasound examination was performed in 866 pregnancies at 6-10 weeks of gestation. The relation of demographic data and ultrasound findings at the time of the initial assessment to subsequent fetal loss was examined. RESULTS: In the 668 singleton pregnancies with live fetuses and complete follow-up there were 50 (7.5%) fetal losses. The incidence of fetal loss increased significantly with maternal age and decreased with gestation. In the pregnancies resulting in fetal loss, compared to those with live births, the incidence of vaginal bleeding and cigarette smoking was higher, the fetal heart rate was significantly lower and the gestation sac diameter was smaller but the yolk sac diameter was not significantly different. CONCLUSION: In pregnancies with a live fetus at 6-10 weeks' gestation the rate of subsequent fetal loss is related to maternal age, gestation, cigarette smoking, history of vaginal bleeding and the ultrasound findings of small gestation sac diameter and fetal bradycardia, relative to crown-rump length. PMID- 14528472 TI - First-trimester uterine artery blood flow and birth weight. AB - OBJECTIVES: To determine reference values for first-trimester uterine artery resistance index (RI) in healthy pregnant women with uncomplicated pregnancies and to investigate the relationship between uterine artery Doppler indices and birth weight. METHODS: This was a cross-sectional study of 265 consecutive pregnant women attending routine ultrasound examination at 11-14 weeks' gestation. Both uterine arteries were identified using color Doppler ultrasound and the RI was measured. The presence or absence of an early diastolic notch was also noted. Pregnancy outcomes were obtained from the delivery suite database and birth weight was expressed as Z-scores. RESULTS: The 5(th), 50(th) and 95(th) centiles for uterine artery RI between 11 and 14 weeks' gestation were 0.53, 0.71 and 0.85, respectively. Complete pregnancy outcome data were available for 246 fetuses. There was a significant negative correlation between birth-weight Z scores and first-trimester uterine artery mean RI (r = - 0.219, P = 0.001). The difference in birth weight between fetuses with absent and those with bilateral diastolic notches was also significant (P < 0.001). Multiple regression analysis demonstrated that both mean RI (standardized regression coefficient = - 0.14, P = 0.039) and uterine artery notching (standardized regression coefficient = - 0.17, P = 0.017) contributed independently towards the prediction of birth-weight Z score. CONCLUSIONS: A significant negative correlation exists between birth weight and first-trimester uterine artery Doppler parameters, a reliable and non invasive method of examining uteroplacental perfusion. The value of first trimester uterine artery Doppler as a prognostic screening tool, either in isolation or in conjunction with maternal biochemistry, remains to be determined. PMID- 14528473 TI - Fetal blood sampling in twin pregnancies. AB - OBJECTIVE: To evaluate the role of fetal blood sampling for prenatal diagnosis in twin pregnancies. METHODS: This was a retrospective study of 84 twin pregnancies that had undergone fetal blood sampling between the years 1977 and 2000. These results were compared with those from a similar study on amniocentesis and chorionic villus sampling. RESULTS: Miscarriage occurred in 3.6% of cases after fetal blood sampling. The total fetal loss rate was 13.6% and the fetal loss that could be attributed to the procedure was 8.2%. The procedure-related fetal loss rate was dependent on the indication for fetal blood sampling. The rates of preterm delivery at 28 weeks and neonatal mortality were 16.9%, 28.3%, 10.9%, 3.1% and 7.7%, respectively. CONCLUSION: Fetal blood sampling in twin pregnancies carries higher risks for the fetus than do amniocentesis and chorionic villus sampling. However, it remains useful in evaluating the fetal condition in special circumstances such as hydrops and infection. PMID- 14528474 TI - Spatio-temporal image correlation (STIC): new technology for evaluation of the fetal heart. AB - Spatio-temporal image correlation (STIC) is a new approach for clinical assessment of the fetal heart. It offers an easy to use technique to acquire data from the fetal heart and to aid in visualization with both two-dimensional and three-dimensional (3D) cine sequences. The acquisition is performed in two steps: first, images are acquired by a single, automatic volume sweep. Second, the system analyzes the image data according to their spatial and temporal domain and processes an online dynamic 3D image sequence that is displayed in a multiplanar reformatted cross-sectional display and/or a surface rendered display. The examiner can navigate within the heart, re-slice, and produce all of the standard image planes necessary for a comprehensive diagnosis. The advantages of STIC for use in evaluation of the fetal heart are as follows: the technique delivers a temporal resolution which corresponds to a B-mode frame rate of approximately 80 frames/s; it provides the examiner with an unlimited number of images for review; it allows for correlation between image planes that are perpendicular to the main image acquisition plane; it may shorten the evaluation time when complex heart defects are suspected; it enables the reconstruction of a 3D rendered image that contains depth and volume which may provide additional information that is not available from the thin multiplanar image slices (e.g. for pulmonary veins, septal thickness); it lends itself to storage and review of volume data by the examiner or by experts at a remote site; it provides the examiner with the ability to review all images in a looped cine sequence. PMID- 14528475 TI - Spatio-temporal image correlation (STIC): a new tool for the prenatal screening of congenital heart defects. AB - OBJECTIVE: To assess the feasibility and capability of STIC acquisition, performed by a general obstetrician performing antenatal ultrasound, to visualize fetal cardiac structures in women undergoing routine obstetric ultrasound examination, in order to obtain information to confirm normality of the fetal heart during intrauterine life. METHODS: This was a prospective study of one hundred fetuses with echocardiographically confirmed normal hearts and no extracardiac anomalies with gestational ages ranging between 18 and 37 weeks. A general obstetrician was invited to acquire the STIC volumes. The four-chamber view was obtained as a starting point. A standard 7.5-s acquisition time and 30 degrees angle of acquisition were used and the resulting STIC was stored for later offline analysis by a fetal echocardiologist. For each patient, the stored STIC data were first evaluated by sweeping from the initial acquisition plane, in the caudal direction and then cranially, zooming, slowing or stopping the cardiac motion to visualize views and structures. If a structure or view was rated as inadequate or not identifiable, a multiplanar three-dimensional (3D) examination of the STIC volume was taken in order to try to visualize it adequately. The rates obtained using just the STIC sweeps were compared independently, and then the 3D multiplanar study was added. RESULTS: STIC acquisition was possible in all cases. The mean time required for STIC acquisition was 7.5 min. A complete cardiac examination according to the set criteria was achieved in 94.2% (95% confidence interval (CI), 90-99) of cases. We obtained a 94.2% success rate of visualizing different structures and views of the fetal heart using the STIC sweep alone (95% CI, 90-99) and 96.2% adding 3D multiplanar examination (95% CI, 92-100). CONCLUSION: STIC acquisition of the fetal heart is feasible with high success rates in visualization of the principal connections. The STIC data acquired by a general obstetrician can subsequently be used by a fetal echocardiologist for prenatal confirmation of normal cardiac structure or exclusion of major cardiac malformations. PMID- 14528476 TI - Impaired dynamic cerebral autoregulation in eclampsia. AB - Eclampsia is frequently associated with brain edema, cerebral infarction or hemorrhage. Its underlying cerebrovascular pathophysiology is still poorly understood. We examined cerebral autoregulation by a non-invasive multimodal assessment in a 28-year-old primaparous woman with postpartal eclampsia. Transcranial Doppler sonography showed considerably increased cerebral blood flow velocity (CBFV) of all basal cerebral vessels. Magnetic resonance imaging demonstrated multifocal vasogenic brain edema. Using transfer function analysis, a severely decreased phase shift between respiratory-induced 0.1-Hz oscillations of arterial blood pressure and CBFV was observed, indicating substantial disturbance of dynamic cerebral autoregulation (DCA). In contrast, CO(2) vasomotor reactivity of the right middle cerebral artery was only slightly reduced. We therefore assume that the cerebral arteriolar dysfunction in eclampsia leads primarily to an impairment of the autoregulatory mechanism that is followed by different degrees of arteriolar vasodilation. Because of its probably high sensitivity to hemodynamic disturbances, assessment of DCA might be of great value in early pre-eclampsia for risk prediction of cerebral arteriopathy and eclampsia. PMID- 14528477 TI - Prenatal diagnosis of ranula at 21 weeks of gestation. AB - A fetal sublingual cystic lesion was diagnosed by routine prenatal ultrasonography at 21 weeks of gestation and followed up until term in a tertiary care center. Fetal growth was normal as was the amniotic fluid volume. Ex utero intrapartum treatment was performed and the cyst was aspirated to allow breathing and swallowing during planned Cesarean section. The cyst was totally excised when the newborn was 27 days of age and histological examination revealed a mucous cyst of the mouth floor. PMID- 14528478 TI - Bony fragments in the uterus: an association with secondary subfertility. AB - Bone within the uterine cavity is an unusual finding in women with secondary subfertility, and is usually associated with a past history of termination of pregnancy. The etiology is unknown, but theories include retained fetal bone and osseous metaplasia of endometrial tissue. We describe the cases of three subfertility patients, all with a history of surgical termination of pregnancy. Each patient underwent a hysteroscopy after highly echogenic foci were seen in the uterus on transvaginal ultrasound examination. During hysteroscopy, several coral-like bony fragments were seen and removed by sharp curettage. On histological examination, these fragments were found to be mature, necrotic bone. This case report confirms the importance of routine baseline evaluation of the endometrium in subfertile women with a history of termination of pregnancy. PMID- 14528479 TI - Uterine intramural bone after mid-trimester termination of pregnancy may not affect fertility: a case report. AB - We present a case of a 27-year-old asymptomatic woman, gravida 2 para 0 abortus 2, diagnosed with uterine intramural fetal bone 30 days after a mid-trimester termination of pregnancy (TOP) by dilatation and evacuation (D&E). On ultrasound part of a fetal spine was seen within the right lateral isthmocervical area, adjacent to the descending branch of the uterine artery. Within 4 months after TOP the patient conceived again. This case illustrates the risk of myometrial penetration during mid-trimester TOP by D&E. Removal of intramural bony fragments may not be needed in an asymptomatic patient, as their presence does not seem to compromise fertility. PMID- 14528480 TI - Acardiac twin: a systematic review of minimally invasive treatment modalities. AB - This review of the literature aimed to determine pregnancy outcomes after minimally invasive treatment for occluding vascular supply to acardiac twins. A MEDLINE search was performed until 2002 using the terms 'acardiac', 'acardius', 'twin reversed arterial perfusion (TRAP) sequence' and 'chorangiopagus parasiticus' and 207 articles published in the English-language literature were identified. These articles and the corresponding cited references in English were reviewed. We identified 32 reports involving 74 cases of acardiac twin treated by minimally invasive techniques. Seventy one cases were included for analysis including 40 treated by cord occlusion and 31 by intrafetal ablation. Cord occlusion was first attempted by embolization (n = 5), cord ligation (n = 15), laser coagulation (n = 10), bipolar diathermy (n = 7) and monopolar diathermy (n = 3). Intrafetal ablation was performed by alcohol (n = 5), monopolar diathermy (n = 9), interstitial laser (n = 4) and radiofrequency (n = 13). The overall median gestational ages at treatment and delivery were 21 (range, 14-28) weeks and 36 (range, 19-42) weeks, respectively, with a median treatment-delivery interval of 13 (range, 0-25) weeks. The overall pump twin survival rate was 76% (n = 54). Intrafetal ablation was associated with later median gestational age at delivery (37 vs. 32 weeks, P = 0.04) and higher median treatment-delivery interval (16 vs. 9.5 weeks, P = 0.02) compared with cord occlusion techniques. It was also associated with a lower technical failure rate (13% vs. 35%, P = 0.03), lower rate of premature delivery or rupture of membranes before 32 weeks (23% vs. 58%, P = 0.003) and higher rate of clinical success (77% vs. 50%, P = 0.02) than cord occlusion techniques. There were no statistically significant differences in terms of outcome between the subgroups of fetoscopic- or ultrasound-guided cord occlusion techniques. This review suggests that intrafetal ablation is the treatment of choice for acardiac twins because it is simpler, safer and more effective when compared with the cord occlusion techniques. PMID- 14528481 TI - The conservative management of early pregnancy complications: a review of the literature. AB - Early pregnancy complications include miscarriage, ectopic pregnancies, adnexal masses and pregnancies of unknown location. In this review, we evaluate the role of conservative management in these complications. We also evaluate the role of transvaginal sonography for diagnosis, treatment and follow up. When managing women with early pregnancy complications, it is important to recognise the normal sonographic milestones in the first trimester. Understanding the pattern of serum human chorionic gonadotropin change in early normal pregnancy and the correlation between low serum progesterone levels and the spontaneous resolution of a pregnancy are also important concepts. Guidelines for assessing suitability for conservative management should be strictly governed by evidence based protocols. Women can then be offered expectant or medical treatment as appropriate. PMID- 14528482 TI - Ultrasound visualization of fetal membrane detachment at the uterine cervix: the 'moon sign'. PMID- 14528483 TI - The 'daughter cyst sign': a sonographic clue to the diagnosis of fetal ovarian cyst. PMID- 14528484 TI - Three-dimensional transvaginal ultrasound provides clearer delineation of myometrial invasion in a patient with endometrial cancer and uterine leiomyoma. PMID- 14528485 TI - Congenital high airway obstruction syndrome successfully managed with ex-utero intrapartum treatment. PMID- 14528487 TI - Oxidation and degradation products of papaverine. Part II[1]: investigations on the photochemical degradation of papaverine solutions. AB - The structure of the final degradation product formed in papaverine solutions in either water or chloroform was found to be a 2, 3, 9, 10-tetramethoxy-12-oxo-12H indolo [2, 1-a] isoquinolinylium salt (a dibenzo [b, g] pyrrocolonium derivative). Its formation from papaverine oxidation products that is papaverinol, papaveraldine, and papaverine-N-oxide chloroform solutions under the influence of UV light, was investigated and possible reaction pathways are discussed. PMID- 14528489 TI - Geometrically constrained analogues of N-benzylindolylglyoxylylamides: [1, 2, 4]triazino[4, 3-a]benzimidazol-4(10H)-one derivatives as potential new ligands at the benzodiazepine receptor. AB - A series of 3-benzylamino-and 3-arylalkylaminocarbonyl [1, 2, 4]triazino [4, 3 a]benzimidazoles 1-12 were synthesized and biologically assayed as geometrically constrained analogues of N-benzylindolylglyoxylylamides II, which are high affinity ligands at the benzodiazepine receptor (BzR). The intermediate 3 ethoxycarbonyl [1, 2, 4]triazino [4, 3-a]benzimidazol-4(10H)-one 14 and its N(10) methyl analogue 15 closely related to 3-alkoxycarbonyl-beta-carbolines I were also investigated. The title compounds exhibited a lower affinity compared with the corresponding indolylglyoxylylamide derivatives II. Attempts were made to rationalize these results taking into account the possible tautomeric equilibria involving these ligands. PMID- 14528488 TI - Investigations of new pyridazinone derivatives for the synthesis of potent analgesic and anti-inflammatory compounds with cyclooxygenase inhibitory activity. AB - In this study we describe the synthesis of two novel 4-phenyl-and 4-(2 chlorophenyl)-6-(5-chloro-2-oxo-3H-benzoxazol-7-yl)-3(2H)-pyridazinone derivatives (compounds 8a and b) and their testing as inhibitors of cyclooxygenases (COX-1 and COX-2). Both compounds inhibited COX-1 (by 59 % and 61 % for compounds 8a and 8b respectively and COX-2 (by 37 % and 28 % for compounds 8a and 8b respectively) at a concentration of 10 microM. Furthermore, we tested the analgesic and anti-inflammatory activities of the synthesized compounds in vivo by using the p-benzoquinone-induced writhing test and the carrageenan induced hind paw edema model, respectively. Compounds 8a and 8b showed potent analgesic and anti-inflammatory activities without causing gastric lesions in the tested animals. PMID- 14528490 TI - Synthesis and evaluation of pharmacological activities of 3, 5-dialkyl 1, 4 dihydro-2,6-dimethyl-4-nitroimidazole-3, 5-pyridine dicarboxylates. AB - New analogues of nifedipine, in which the 2-nitrophenyl group at position 4 is replaced by a 1-methyl-5-nitro-2-imidazolyl substituent, were synthesized. The symmetrical dialkyl 1, 4-dihydro-2, 6-dimethyl-4-(1-methyl-5-nitro-2-imidazolyl) 3, 5-pyridinedicarboxylates were prepared by a classical Hantzsch condensation. The asymmetrical analogues were synthesized using a procedure reported by Iwanami that involved the condensation of alkylacetoacetate with methyl-, ethyl- or isopropyl3-aminocrotonate and 1-methyl-5-nitroimidazole-2-carboxaldehyde. Calcium channel antagonist activities were determined in vitro using a guinea pig ileum longitudinal smooth muscle (GPILSM)assay. Many compounds exhibited superior, or equipotent, calcium antagonist activity (IC(50) = 10(-10) to 10(-13) M range) relative to the reference drug nifedipine (IC(50) = 1.09 +/- 0.12 x 10(-11) M). Antinociceptive effects of some compounds were evaluated by the mouse tail-flick assay in vivo. Results demonstrate that some of the compounds were active as an antinociceptive. PMID- 14528491 TI - Synthesis and effects on the COX-1 and COX-2 activity in human whole blood ex vivo of derivatives containing the [1]benzothienol-[3, 2-d]pyrimidin-4-one heterocyclic system. AB - Methyl and phenyl derivatives containing the [1]Benzothieno [3, 2-d]pyrimidin-4 one system have been synthesized and tested as inhibitors of COX-1 and COX-2 activities in human whole blood (HWB) ex vivo; all compounds turned out to be weak inhibitors of COX-1 activity, as deduced from the TXB(2) (thromboxane B) generation; the acid phenyl derivative 11 b was an interesting inhibitor of COX-2 activity, as deduced from the PGE(2) (prostaglandine E) generation. PMID- 14528493 TI - Development of new methodologies for the mass spectrometry study of bioorganic macromolecules. AB - In recent years, mass spectrometry has been increasingly used for the analysis of various macromolecules of biological, biomedical, and biochemical interest. This increase has been made possible by two key developments: the advent of electrospray ionization (ESI) and matrix-assisted laser desorption ionization (MALDI) sources. The two new techniques produce a significant increase in mass range and in sensitivity that led to the development of new applications and of new analyzer designs, software, and robotics. This review, apart from the description of the status of mass spectrometry in the analysis of bioorganic macromolecules, is mainly devoted to the illustration of the more recent promising techniques and on their possible future evolution. PMID- 14528494 TI - Gas-phase ion thermochemistry in organometallic systems. AB - This review represents a general overview on the determination of bond energies in gas-phase organometallic systems. The paper focuses on articles published in the last 7 years, whose main scope was to experimentally measure metal ion-ligand binding energies. Therefore, studies of reactivity and characterization of ion complexes are not included, even if some of them display bond energies among the data measured. Bond energies are presented according to the metal ion complexed, in increasing order of atomic number. Periodic trends of binding energies and correlations among the various ligand are discussed. A brief summary of the most used techniques specifically devoted to gather bond energies is also provided. PMID- 14528495 TI - Non-specific, on-probe cleanup methods for MALDI-MS samples. AB - High concentrations of contaminants such as salts and surfactants are often present in biological samples to solubilize or stabilize analytes such as proteins. Unfortunately, the presence of those contaminants often precludes direct analysis by MALDI-MS. Selective adsorption of analytes directly on modified MALDI probes, followed by rinsing to remove contaminants, overcomes this problem. This review focuses on various modifications of MALDI probes to allow the adsorption of proteins and DNA, even in a large excess of salt or surfactant. Interfaces deposited on the MALDI probes to adsorb analytes include films of commercial polymers, thin layers of matrix crystals, self-assembled monolayers, and ultrathin polymer films. Hydrophobic and ionic interactions both effect analyte adsorption on those interfaces, and patterned interfaces allow the concentration and purification of analyte molecules. PMID- 14528497 TI - Another DMARD option in rheumatoid arthritis? PMID- 14528498 TI - Clinical research into alternative and complementary therapies: how do we tell if the glass is half empty or half full? PMID- 14528499 TI - Rheumatic disease in Han Chinese. What have we learned from 19 years of epidemiological study? PMID- 14528500 TI - Where there's smoke there's fire: the silicone breast implant controversy continues to flicker: a new disease that needs to be defined. PMID- 14528501 TI - Unified theory of the origins of erosive arthritis: conditioning as a protective/directing mechanism? AB - OBJECTIVE: To validate the western Tennessee River limits of the originally described rheumatoid arthritis (RA) catchment area and to assess the possibility that absence of tuberculosis allowed the original development of RA. The hypothesis that RA was related to tuberculosis was once a driving force in treatment approach. RA initially was very limited in geographic distribution, in contrast to tuberculosis. Classical tubercular lesions were not observed in the rheumatoid catchment area in ancient times. Similarities between clinical and radiologic manifestations of spondyloarthropathy (SpA) and adjuvant arthritis raised the possibility of a potential conditioning role for occurrence of nonrheumatoid erosive arthritis. METHODS: Skeletal samples from ancient RA catchment and non-catchment areas were compared for frequency of tubercular relatable pathologies. RESULTS: Tubercular-relatable osseous pathologies were found only outside the rheumatoid catchment area (p < 0.0001). The original RA catchment area was confirmed not to extend beyond the western portion of the Tennessee River. CONCLUSION: There is an inverse relationship between occurrence of tuberculosis and RA in the Archaic and Early Woodland periods of North America. The virtually universal presence of tuberculosis in contiguous Amerindian populations contrasts dramatically with its absence in the ancient catchment area for RA. Conversely, SpA and tuberculosis do occur in the same populations. Tuberculosis may represent a conditioning agent for development of SpA, but at least potentially provides protection against development of RA. PMID- 14528502 TI - Risk factors for incident self-reported arthritis in a 20 year followup of the Alameda County Study Cohort. AB - OBJECTIVE: This longitudinal study examined the following variables as possible risk factors for self-reported arthritis: age, sex, race, body mass index (BMI), depressive symptoms, leisure-time physical activity, cigarette use, alcohol, hypertension, diabetes mellitus, education, income, and hard physical work. METHODS: Altogether, 1149 women and 964 men from the Alameda County Study Cohort without self-reported arthritis in 1974 were assessed for incident self-reported arthritis in 1994. RESULTS: In a multivariate model, the following variables were associated with increased odds of incident arthritis: increasing age (age 45-49, odds ratio 2.00, 95% confidence interval 1.40-2.85; age 50+, OR 3.13, 95% CI 2.32 4.22), BMI for women only (4th quintile, OR 1.65, 95% CI 1.05-2.60; 5th quintile, OR 1.88, 95% CI 1.19-2.95), female sex (OR 1.48, 95% CI 1.20-1.83), and >/= 5 depressive symptoms (OR 1.53, 95% CI 1.12-2.10). Leisure-time physical activity in the highest quartile was protective (OR 0.69, 95% CI 0.51-0.95). All other factors were not associated with arthritis. CONCLUSION: This study indicates that depressive symptoms, as well as age, sex, and BMI, are independent risk factors for arthritis. This is the first longitudinal population based study to examine and establish that prior depressive symptoms are a risk factor for arthritis. PMID- 14528503 TI - Should tetracycline treatment be used more extensively for rheumatoid arthritis? Metaanalysis demonstrates clinical benefit with reduction in disease activity. AB - OBJECTIVE: To compare the effectiveness of tetracycline antibiotics versus control (placebo or conventional treatment) in rheumatoid arthritis (RA) for the reduction of disease activity as defined by American College of Rheumatology criteria. METHODS: We searched Medline (1966-February 2002), Embase (1980 February 2002), and the Cochrane Controlled Trials Register (Issue 1, 2002 Cochrane Library). Reference lists of published trials were searched by hand for further identification of published reports and presentations at scientific meetings. Randomized controlled trials comparing tetracyclines to control (placebo or conventional disease modifying antirheumatic therapy) were selected for inclusion if at least one of the following outcomes was reported: tender joint count (TJC), swollen joint count, patient pain score by visual analog scale, patient global assessment of disease activity, physician global assessment of disease activity, eosinophil sedimentation rate (ESR) and C-reactive protein (CRP), joint space narrowing and erosions, adverse events, and quality of life as measured by the Health Assessment Questionnaire. Subjects were required to have RA as defined by the 1987 ARA criteria. RESULTS: Ten randomized controlled trials including 535 individuals were reviewed. Only 3 trials were considered high quality; elements of bias could not be excluded in the remainder. Tetracyclines, when administered for > or = 3 months, were associated with a significant reduction in disease activity in RA as follows: for TJC, standardized mean difference (SMD) = -0.39, 95% CI -0.74, -0.05; and for acute phase reactants, ESR, SMD = -8.96, 95% CI -14.51, -3.42. The treatment effect was more marked in the subgroup of patients with disease duration < 1 year who were seropositive. There was no absolute increased risk of adverse events associated with tetracyclines: absolute risk difference = 0.10, 95% confidence interval (CI) 0.01, 0.21. No beneficial effect was seen on radiological progression of disease: for erosions, SMD = 0.17, 95% CI -0.29, 0.64. In addition, subgroup analysis excluding trials with doxycycline showed that minocycline alone had a greater effect on reduction of disease activity: for TJC, SMD = -0.69, 95% CI -0.89, 0.49; and for ESR, SMD = -10.14, 95% CI -14.72, -5.57. CONCLUSION: Tetracyclines, in particular minocycline, were associated with a clinically significant improvement in disease activity in RA with no absolute increased risk of side effects. Unfortunately, the information available was inadequate to allow a detailed analysis of individual side effects in the studies. Further research is warranted to compare these agents to newer disease modifying drugs for comparable safety, efficacy, and cost-effectiveness. PMID- 14528504 TI - Intravenous human recombinant tumor necrosis factor receptor p55-Fc IgG1 fusion protein, Ro 45-2081 (lenercept): results of a dose-finding study in rheumatoid arthritis. AB - OBJECTIVE: To determine the optimal dose regimen of intravenous (IV) Ro 45-2081 (lenercept), a tumor necrosis factor receptor p55-Fc IgG1 fusion protein, in patients with active rheumatoid arthritis (RA) METHODS: In a double-blind, placebo-controlled, parallel-group, multicenter trial, adult patients with long standing active RA stabilized on conventional therapy were randomly assigned to receive 3 IV infusions, one every 4 weeks, of one of the following: (a) placebo, (b) lenercept 0.01 mg/kg (maximum 1 mg), (c) lenercept 0.05 mg/kg (maximum 5 mg), (d) lenercept 0.2 mg/kg (maximum 20 mg), or (e) lenercept 0.5 mg/kg (maximum 50 mg). The material utilized in the study had a lower relative bioavailability [lower area under the time-concentration curve (AUC) per mg infused] than that used in a recent similar trial. Efficacy variables included change from baseline in number of swollen joints and tender joints, scores on physician and patient assessments of disease activity, and patient assessment of pain. RESULTS: Patients treated with lenercept exhibited improvement as early as one day after the first IV infusion. The treatment benefit, however, was modest, maximized by 2 weeks and then diminished or vanished as non-neutralizing anti-lenercept antibody concentrations increased. The majority of adverse experiences were mild or moderate and not considered related to study drug. CONCLUSION: Our results showed that lenercept administered by IV infusion every 4 weeks is well tolerated, but only transiently effective in patients with long-standing RA, likely due to both the low relative bioavailability of the material used in the study and the formation of non-neutralizing anti-lenercept antibodies. PMID- 14528505 TI - Preterm deliveries in women with systemic lupus erythematosus. AB - OBJECTIVE: To compare the clinical, laboratory, and demographic variables of women in our clinic with systemic lupus erythematosus (SLE) who have had a pregnancy resulting in a live birth and identify any correlations with either term or preterm delivery. METHODS: Pregnancies in women with SLE from 1999 to 2001 were retrospectively reviewed. We recorded demographic data, disease activity (SLE Disease Activity Index, SLEDAI), obstetric history, prednisone dosage, other medications taken during pregnancy, history of renal disease, and autoantibody status [including antinuclear antibody, anti-DNA, anticardiolipin IgG (aCL), and lupus anticoagulant (LAC)]. Preterm delivery was defined as gestational age at delivery < 37 weeks. We performed a literature survey using PubMed and the key words SLE, pregnancy, and outcome. RESULTS: Of the 72 pregnancies, 28 (38.9%) resulted in preterm deliveries. There were no significant differences in any demographic or disease variables measured comparing term versus preterm delivery groups. More women in the preterm group were taking > or = 10 mg/day prednisone during their pregnancy (50.0% vs 22.2%; p = 0.028), and the mean dose was significantly higher than the term group taking > or = 10 mg/day (24.8 vs 16.7 mg/day; p = 0.047). There was a higher prevalence of women with aCL IgG in the preterm group (p = 0.023). The mean weeks gestation was shorter for women positive for aCL IgG compared to the group negative for aCL (34.9 +/- 4.4 vs 37.5 +/- 3.2 weeks, respectively; p = 0.032). There was no difference in second trimester disease activity between the term and preterm groups (33.3% and 36.4% of each group had a SLEDAI of 0). However, significantly more women in the term group received no medication during their pregnancies compared to women in the preterm group (20.0% vs 0.0%; p = 0.031). CONCLUSION: The rates of preterm deliveries, premature rupture of membranes, intrauterine growth restriction, and aPL in SLE pregnancies vary considerably in published reports, most of which are retrospective analyses. Our rates closely approximate the median values for all measures. We found preterm deliveries to be associated with disease activity (as determined by the use of any medication throughout pregnancy vs no medication, and prednisone dose > or = 10 mg/day) and the presence of aCL IgG but not LAC. Our results suggest that inactive disease rather than controlled disease at the onset of pregnancy may be the determining factor in extending SLE pregnancies to full term, thereby decreasing maternal and fetal morbidity. PMID- 14528506 TI - Subclinical disease activity in systemic lupus erythematosus: immunoinflammatory markers do not normalize in clinical remission. AB - OBJECTIVE: To correlate various laboratory measures in systemic lupus erythematosus (SLE) with the British Isles Lupus Assessment Group (BILAG) disease activity index, to search for organ-specific laboratory patterns and to compare with a control population. METHODS: A cohort of 57 Caucasian outpatients fulfilling the American College of Rheumatology criteria for SLE and a control population of 17 patients admitted for coronarography were examined. Disease activity was assessed with BILAG index. Plasma samples were investigated for sCD44, interleukin 6 (IL-6), IL-10, IL-12, tumor necrosis factor-a (TNF-a), soluble TNF receptor-55 (sTNFR-55), sTNFR-75, IL-1-receptor antagonist, soluble intercellular adhesion molecule (sICAM), soluble vascular cell adhesion molecule (sVCAM), E-selectin, and neopterin as well as for C3, C4, dsDNA, and other conventional indicators. RESULTS: Thirty-nine patients had inactive disease (total BILAG score < or = 5), 18 patients had active SLE. Surprisingly, except for C-reactive protein (p < 0.001), no statistically significant difference of the laboratory indicators was found between patients with active and those with inactive SLE. However, there was a significant difference between SLE patients and controls for sTNFR-75 (p < 0.008). We found significant correlations between laboratory markers and some BILAG organ system scores, such as between IL-1ra and the musculoskeletal score (p < 0.003) and between sTNFR-55/sTNFR-75 and renal BILAG (p < 0.001, p < 0.004, respectively). Significant nonparametric correlations were revealed between C3 and C4 (p < 0.0001), and between sTNFR-75 and dsDNA, neopterin, sVCAM, sICAM and sTNFR-55 concentrations (p < 0.0001 for all), and between sTNFR-75 and IL-1ra (p < 0.006). CONCLUSION: Patients with SLE in clinical remission show ongoing systemic immunoinflammatory activity measured with a variety of cytokines, adhesion molecules, and other inflammatory markers. This indicates that laboratory measures may provide qualitatively different additional information to validated disease activity indexes such as the BILAG. Different laboratory markers correlate with disease activity in different organ systems. This suggests differences in pathogenic mechanisms in SLE depending on the organ system involved. PMID- 14528507 TI - Correlation of prolactin serum concentrations with clinical activity and remission in patients with systemic lupus erythematosus. Effect of conventional treatment. AB - OBJECTIVE: The role of prolactin (PRL) in the pathogenesis of systemic lupus erythematosus (SLE) is controversial. The effect of conventional treatment (steroids, antimalarials, immunosuppressor drugs) on PRL concentrations is unclear. We investigated correlation of PRL levels with lupus activity in patients at entry and after 6 months of conventional treatment. METHODS: We studied 43 female patients with active SLE, who were divided in 2 groups; Group 1: 16 patients with minor organ involvement (cutaneous and articular involvement), and Group 2: 27 patients with major organ involvement (glomerulonephritis). Controls were 36 healthy individuals. PRL levels were determined by an immunoradiometric assay at entry and after 6 months of treatment. PRL levels were correlated with SLE Disease Activity Index (SLEDAI) score. RESULTS: Mild hyperprolactinemia (HPRL, 20-40 ng/ml) was found in 30/43 (69.7%) SLE patients. After 6 months of treatment a reduction in PRL levels was found in both groups: Group 1: 24.3 +/- 10.8 to 16.96 +/- 10.87 ng/ml (p < 0.001); and Group 2: 23.6 +/- 5.7 to 12.07 +/- 11.13 ng/ml (p < 0.001). The SLEDAI score also decreased after treatment: Group 1: 16.5 +/- 5.9 to 2.1 +/- 1.3 (p < 0.001); Group 2: 16.8 +/- 5.4 to 1.6 +/- 1.4 (p < 0.001). At entry and after treatment, a significant correlation between PRL levels and SLEDAI score was found in all patients (r = 0.4946, p = 0.0007, and r = 0.9086, p = 0.0001, respectively). CONCLUSION: HPRL was associated with SLE disease activity. Conventional immunosuppressive therapy decreased PRL levels in direct correlation with decreased SLE activity. This finding emphasizes that PRL may play a role in the pathogenesis and clinical expression of SLE. PMID- 14528508 TI - Cross-sectional evaluation of YKL-40 serum concentrations in patients with systemic sclerosis. Relationship with clinical and serological aspects of disease. AB - OBJECTIVE: To investigate the behavior of serum YKL-40 in a cohort of patients with systemic sclerosis (SSc). METHODS: Forty SSc patients (35 women, 5 men) were investigated for serum YKL-40, soluble interleukin 2 receptor alpha (sIL-2Ra; by ELISA), von Willebrand factor (vWF; ELISA), and aminoterminal propeptide of type III procollagen (PIIINP; radioimmunoassay) concentrations. Erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were evaluated. Skin and organ system involvement were scored according to the Medsger organ/system severity scale. RESULTS: Serum YKL-40 in SSc patients (mean 132.9 ng/ml; median 75.5, 95% CI 87.8-175) was significantly higher than in controls (mean 66.6 ng/ml; median 52, 95% CI 54.6-78.6). Fourteen patients had levels > 135 ng/ml (cutoff value) with a mean of 264.7 +/- 160 ng/ml. Serum YKL-40 values were found to be more frequently increased in patients with arthralgias/arthritis (18 cases) than in patients without such features (4/22) (p = 0.021). Significant differences were found comparing serum YKL-40 concentrations in the patients with or without joint involvement (median 138 vs 57.5 ng/ml, respectively; p = 0.007). Serum YKL-40 levels correlated with the joint involvement severity score (p = 0.018) and sIL-2Ra levels (p < 0.001). No differences were found with any therapeutic regimen. CONCLUSION: This preliminary study shows that YKL-40 serum levels are increased in SSc and that they are correlated with sIL-2Ra and joint involvement, suggesting a relationship with cartilage and/or fibroblast activity. PMID- 14528509 TI - D6S439 microsatellite identifies a new susceptibility region for primary Sjogren's syndrome. AB - OBJECTIVE: To examine genetic variations in the region surrounding loci of the major histocompatibility complex, and to investigate the probable location of a new candidate region on the short arm of chromosome 6 predisposing to primary Sjogren's syndrome (SS). METHODS: We conducted an association study and positional candidate gene approach by microsatellite analysis. Five polymorphic microsatellite markers, D6S273, D6S439, D6S1645, D6S291, and DS61019, spanning the region 6p21.3, and establishing particular landmarks to discriminate between the human leukocyte antigen class II and tumor necrosis factor-a loci, were genotyped by polymerase chain reaction technique. RESULTS: A total of 64 patients with primary SS and 120 matched controls were examined. There was no genetic stratification among cases and controls. Genotype distribution analysis disclosed a significantly higher number of homozygotes for D6S439 locus in patients than in controls [odds ratio (OR): 3, 95% confidence interval (CI): 1.46-6.14, p = 0.004]. Confirmation of this homozygosity was established by the gene correlation intra-locus test (Fis value = +0.233, p = 0.0007). Allele D6S439*274 was associated to disease (OR: 3, 95% CI: 1.35-6.65, p = 0.006, pc = 0.04). Among patients, no significant linkage disequilibrium (LD) value was found between the studied microsatellites and TAP, HLA-DRB1, or HLA-DQB1 loci. In controls, there was LD between D6S1645 and D6S291 loci. CONCLUSION: Our results indicate that D6S439 microsatellite defines a new susceptibility region for primary SS, independent of LD with TAP and HLA DQ/DR. These findings might imply that a gene surrounding this location is causally related to the disease. PMID- 14528510 TI - Prevalence of antibodies against alpha-fodrin in Sjogren's syndrome: comparison of 2 sets of classification criteria. AB - OBJECTIVE: To compare the prevalence of antibodies against alpha-fodrin in Sjogren's syndrome (SS) classified according to San Diego and European Community Study Group (ESG) criteria. METHODS: The prevalence and mean concentrations of IgA and IgG autoantibodies against alpha-fodrin were determined and compared in patients with SS classified either according to San Diego criteria or to criteria of the ESG by ELISA. RESULTS: IgA antibodies against alpha-fodrin were detected in 88% and IgG antibodies against alpha-fodrin in 64% and either of these antibodies in 93% of 85 patients classified according to San Diego criteria. Antibodies against Ro were present in 38% of these sera. IgA antibodies against alpha-fodrin were detected in 61%, IgG antibodies against alpha-fodrin in 51%, and any of these antibodies in 73% of 51 patients classified according to the ESG criteria. The mean concentrations of both IgA and IgG antibodies against alpha fodrin that seem to correlate with disease activity were higher in patients classified according to the San Diego criteria. CONCLUSION: Antibodies against alpha-fodrin are detectable in almost all sera obtained from patients with SS classified according to the San Diego criteria and are significantly more prevalent than antibodies against Ro. The lower prevalence of the autoantibodies in patients classified according to the ESG criteria reflects the lower specificity of these criteria for SS. PMID- 14528511 TI - Vascular endothelial growth factor gene polymorphisms in giant cell arteritis. AB - OBJECTIVE: To examine potential associations of vascular endothelial growth factor (VEGF) gene polymorphisms with giant cell arteritis (GCA) and disease expression, in particular in patients with and without ischemic complications. METHODS: We enrolled 92 consecutive patients with biopsy-proven GCA residing in Reggio Emilia, Italy. Two hundred healthy blood donors from the same geographic area were selected as controls. All the GCA patients and controls were genotyped by polymerase chain reaction and allele-specific oligonucleotide techniques for 936 C/T and 634 C/G mutations and for an 18 bp insertion/deletion (I/D) polymorphism in the VEGF promoter region. In vitro release of VEGF by peripheral blood mononuclear cells (PBMC) was investigated by ELISA in controls homozygous for the polymorphisms studied. RESULTS: The carriage rates of the alleles I and C634 were significantly more frequent in GCA patients than in controls (p = 0.025, OR 1.9, 95% CI 1.1-3.1 and p = 0.015, OR 2.1, 95% CI 1.1-3.6, respectively). The distribution of allele T936 was similar in GCA patients and controls. No significant differences in the distribution of the polymorphisms studied were observed in patients with ischemic manifestations compared to those without ischemic manifestations. Lipopolysaccharide (LPS)-stimulated VEGF production by PBMC from controls was higher in II homozygous compared to DD homozygous patients. CONCLUSION: Our data indicate that carriers of C634 and I alleles are associated with susceptibility to developing GCA. PMID- 14528512 TI - Temporal arteritis associated with systemic necrotizing vasculitis. AB - OBJECTIVE: To evaluate the clinical and laboratory characteristics of patients with systemic vasculitis associated with temporal artery involvement. METHODS: From a cohort of 120 patients fulfilling American College of Rheumatology criteria for temporal arteritis, we retrospectively identified 7 patients with systemic necrotizing vasculitis associated with histological temporal arteritis. RESULTS: Among the 7 patients, 2 had classic polyarteritis nodosa, one had unclassified systemic vasculitis, one had Wegener's granulomatosis (WG), and 3 had microscopic polyangiitis. The mean age of the patients was 70.2 years, and cranial symptoms revealed the disease in all but one patient. Temporal arteritis was generally associated with extracephalic manifestations suggestive of systemic vasculitis. Antineutrophilic cytoplasmic antibodies were positive in 3 of the 4 patients with small vessel vasculitis. Pathologically, the main temporal artery was involved in all but one patient, with inflammatory infiltrate of vasa vasorum and adventitia associated in 5 with small tributary involvement. Fibrinoid necrosis was rare, observed in 2 specimens; 2 patients with unclassified systemic vasculitis and WG had a classic giant cell arteritis (GCA) histologic pattern. Only one patient had exclusive involvement of small vessels, surrounding the spared main temporal artery. Muscle biopsies showed histopathological evidence of vasculitis in 2 patients, skin biopsy in one, and vein biopsy in the other. CONCLUSION: Temporal artery involvement in systemic necrotizing vasculitis was generally associated with extracranial clinical features suggestive of systemic vasculitis. Temporal artery biopsy is a simple tool for diagnosis of vasculitis, but the histopathological findings do not always discriminate between necrotizing vasculitis and classic GCA. PMID- 14528513 TI - Contrast medium in power Doppler ultrasound for assessment of synovial vascularity: comparison with arthroscopy. AB - OBJECTIVE: To evaluate the reliability of contrast-unenhanced power Doppler (CUPD) and contrast-enhanced power Doppler (CEPD) ultrasound (US) assessment of synovial vascularity of knee joint synovitis by prospective comparison with the "gold standard," arthroscopy. METHODS: A total of 18 knees of 17 patients with refractory rheumatoid and psoriatic knee joint synovitis were examined by US. Recognition of PD synovial vessel flow and its spatial arrangement in relation to the pannus/cartilage interface (P/CI) or fluid/synovium interface (F/SI) were studied by CUPD- and CEPD-US after a single intravenous bolus of galactosel palmitic acid (Levovist). Arthroscopy video recordings were reanalyzed by computer image analysis to assess synovial vascular marking. CUPD and CEPD flow signal scores were compared with each other and with corresponding vascular marking scores. Using villous vascular marking as reference, CUPD and CEPD sensitivity and specificity were measured. Interobserver variability was evaluated. RESULTS: Compared with the unenhanced PD method, contrast administration increased the PD flow signal score in 13/18 knees (72.2%), allowing increased detection of F/SI PD flow signal configuration (p < 0.018) and of the coexistence of P/CI and F/SI PD imaging (p < 0.0078). With arthroscopy as reference, contrast-enhanced PD was found to be more useful than the unenhanced method, showing more reproducible PD signal scores (p = 0.05 vs p = nonsignificant), as well as higher sensitivity (80% vs 30%), but lower specificity (62% vs 87%), in the recognition of increased vascularity of synovial villi. Interobserver agreement was 100%. CONCLUSION: The prospective comparison with arthroscopy showed the reliability of the CEPD method in synovial vessel recognition and its potential clinical usefulness in assessment of knee joint synovitis. PMID- 14528514 TI - Duplex sonography of the temporal and occipital artery in the diagnosis of temporal arteritis. A prospective study. AB - OBJECTIVE: Evaluation of the diagnostic contribution of color coded duplex sonography (CCDS) of the superficial temporal (STA) and the occipital artery (OCCA) in biopsy-controlled patients suspected of having temporal arteritis (TA). METHODS: Prospective study in 67 patients suspected of having TA who underwent CCDS of the STA in all cases and the occipital arteries if involvement of the OCCA was suspected clinically. The final diagnosis, based on biopsy results in 67 cases and standard criteria, were compared to the ultrasonographic findings to determine their diagnostic contribution. RESULTS: TA was diagnosed in 40 patients, other diseases in 27 patients. In the STA periarterial hypoechogenic tissue, the so-called halo, halo and stenoses, and occlusions were found in 83% of TA patients and 11% of patients with other diseases. In the OCCA, these abnormalities were found in 65% of TA patients and in no patient with other diseases. Taking both STA and OCCA together, halo, stenosis, and widespread abnormalities were found in patients with TA, but not in patients with other diseases. CONCLUSION: CCDS of the STA and OCCA clearly contributes to the diagnosis of TA, with a high rate of perivascular hypoechogenic abnormalities (so called halos) and stenosis and a low rate of these abnormalities in the control patients. However, CCDS cannot differentiate between inflammatory and degenerative artery disease and has spatial resolution limitations. PMID- 14528515 TI - Maternal age and family history are risk factors for ankylosing spondylitis. AB - OBJECTIVE: To investigate prevalence and gender distribution in parents of children with ankylosing spondylitis (AS). METHODS: Family history of AS (parents, uncles, and aunts), maternal age at delivery, and consecutive pregnancy number were assessed in the relatives of 40 Mexican Mestizo patients with definite AS (New York Criteria). RESULTS: We evaluated the family history of AS in 34 families of 40 AS patients; 12 with none, 4 with a paternal history (4 healthy fathers with a brother with AS) (odds ratio, OR, 1.37, p = 0.75), 15 with a maternal history of AS, (15 healthy mothers with a brother with AS) (OR 1.4, p = 0.55), and 3 with both lines (OR 0.84, p = 0.92). In these families AS was more frequent in males (29%) than in females (10%), OR 3.40 (95% confidence interval, CI: 1.43-8.29, p = 0.003). Juvenile onset was more common in the offspring of mothers with family history (72%) (OR 13.0, 95% CI: 1.68-147.48, p = 0.009). The number of first-born children with AS (18%) was similar to the later-born children (23%) (OR 1.37, 95% CI: 0.38-5.31, p = 0.78). The frequency of AS increased when the maternal age at delivery was < or = 30 years (OR 0.20, 95% CI: 0.04-0.75, p = 0.01). CONCLUSION: In Mexican Mestizo patients, there is no correlation between the risk for AS and the gender of the affected parent. However we found an association between juvenile onset and maternal family history with an increased incidence in patients with younger mothers. PMID- 14528516 TI - Factors associated with body function and disability in patients with ankylosing spondylitis: a cross-sectional study. AB - OBJECTIVE: To determine for patients with ankylosing spondylitis (AS) which factors are associated with disability and restricted body function. METHODS: In 1538 patients with AS, occiput-to-wall distance, chest expansion, cervical rotation, finger-floor distance, and modified Schober test were measured cross sectionally, and for each measure the patient's respective percentile was calculated. The mean of the 5 percentiles was summarized in the mobility restriction score (MRS). In addition, patients answered questions on disease progression and completed the Bath AS Functional Index (BASFI) questionnaire. All data were coded into 40 variables, used as independent variables in multiple regression analyses to identify factors associated with BASFI and MRS. RESULTS: Finger-to-floor distance (beta positive, +), cervical rotation (-), time since first AS symptoms (-), age (+), height loss (+), maximum height (-), delay in diagnosis (+), hip replacement (+), regular practice of sports (-), chest expansion (-), sex (worse BASFI in females), exposure to cold and dampness (+), and regular participation in AS group physical exercises (+) were significantly associated with worse BASFI. Significantly associated factors for worse MRS were height loss (+), sex (higher MRS in males), active inflammation of the cervical region (+), age (+), maximum height (-), active inflammation of the hip region (+), involvement of shoulders (+), time since first AS symptoms (+), urethritis ( ), regular practice of sports (-), involvement of feet (-), and hip replacement (+). The models explain 47% of the variance observed for both the BASFI and MRS. CONCLUSION: The amount of variance explained for both BASFI and MRS is rather high. The results apply primarily to groups of patients, but are insufficient to guide clinical decisions in individual patients. These findings contribute to our understanding of factors influencing disability and restriction in body function in AS. PMID- 14528517 TI - Differential effects of FK506 and methotrexate on inflammatory cytokine levels in rat adjuvant-induced arthritis. AB - OBJECTIVE: To investigate the effects of prophylactic and therapeutic treatments with FK506 (tacrolimus), an immunosuppressive drug that specifically inhibits T cell activation, and methotrexate (MTX) on inflammatory cytokines, tumor necrosis factor (TNF)-a, interleukin (IL)-1beta, and IL-6 levels in rat adjuvant-induced arthritis (AIA). METHODS: AIA was induced in female Lewis rats. Arthritis was assessed by hindpaw swelling. TNF-a, IL-1beta, and IL-6 levels in paw extracts were determined by ELISA. To assess the effects on cytokine levels, rats were treated prophylactically with FK506 (3 mg/kg) or MTX (0.1 mg/kg) from day 1 to day 17, and therapeutically with FK506 (5 mg/kg) or MTX (1 mg/kg) from day 15 to day 17 (3-day treatment) or day 15 to 20 (6-day treatment) by oral administration. RESULTS: TNF-a, IL-1beta, and IL-6 levels in paw tissue were found to significantly increase between day 15 and day 21 after adjuvant injection, when the arthritis was in a developed stage. Prophylactic treatment with FK506 and MTX suppressed arthritis and reduced the levels of those inflammatory cytokines. FK506 caused a marked reduction of TNF-a and IL-1beta levels in paw tissue even in short-term (3-day) therapeutic treatment. It reduced all levels of TNF-a, IL-1beta, and IL-6 in paws in 6-day therapeutic treatment. In contrast, therapeutic treatment with MTX affected neither TNF-a or IL-6 levels in paws. MTX reduced IL-1beta levels only in the 6-day treatment. CONCLUSION: FK506 is more effective than MTX in reducing elevated levels of inflammatory cytokines TNF-a, IL-1beta, and IL-6 in established stages of AIA. Our findings suggest that inhibition of T cell activation results in a rapid reduction of inflammatory cytokine levels even after the arthritis is established in AIA. PMID- 14528518 TI - Racial differences in health care utilization among patients with osteoarthritis. AB - OBJECTIVE: Research has identified racial variations in certain aspects of osteoarthritis (OA) related medical care. We compared health services utilization between African American and white veteran outpatients with OA. METHODS: Subjects were 1612 white and 861 African American patients receiving medical care for OA at the Durham VAMC, Durham, NC, USA. Two major components of OA related medical care were examined during a one-year period: physician visits and use of analgesic and antiinflammatory medications. RESULTS: There were no racial differences in overall frequency of OA related physician visits or visits to rheumatologists. About 86% of both African American and white patients were prescribed some analgesic or antiinflammatory medication. There were, however, racial differences in the use of specific drug classes. African Americans were more likely to be prescribed nonselective nonsteroidal antiinflammatory drugs (69% vs 60%), but less likely to be prescribed COX-2 inhibitors (4% vs 7%) and narcotic analgesics (33% vs 40%) than whites (all p < 0.05). African Americans also had a shorter annual mean days' supply for several common medications, including acetaminophen, acetaminophen combined with codeine, and acetaminophen combined with oxycodone (all p < 0.05). CONCLUSION: African Americans and white veterans with OA did not differ substantially in their use of physician services. However, within this equal access health care system that requires minimal co payments for medications, there were racial differences in prescription medication use. These differences may have implications for both quality of pain relief and risk of side effects. PMID- 14528519 TI - Expression of the cartilage derived anti-angiogenic factor chondromodulin-I decreases in the early stage of experimental osteoarthritis. AB - OBJECTIVE: Chondromodulin-I (ChM-I), a cartilage derived anti-angiogenic factor, has been shown to regulate the vascular invasion during endochondral bone formation. We evaluated the expression and localization of ChM-I in articular cartilage during the progression of osteoarthritis (OA) in the rat, and correlated ChM-I expression with the increase in vascular invasion into OA articular cartilage. METHODS: Expression of ChM-I, type II collagen, basic fibroblast growth factor, vascular endothelial growth factor (VEGF), and matrix metalloproteinases MMP-9 and MMP-13 were examined in articular cartilage of intact growing and adult rats and in the surgically induced OA model using in situ hybridization, Western blot analysis, and immunohistochemistry. Co immunostaining for ChM-I and CD-31 was performed to localize ChM-I and neovascularization in articular cartilage at advanced stage of OA. RESULTS: Abundant expression of ChM-I protein was detected in avascular regions of the developing and adult healthy articular cartilage. In early OA, ChM-I expression decreased in the superficial zone of articular cartilage, while levels of proteoglycan and type II collagen were comparable to control. In advanced OA, ChM I expression was reduced in all zones of articular cartilage, and the number of VEGF-expressing cells was increased. Immunohistochemical studies showed that vascular invasion occurred in proximity to chondrocytes with high expression of pro-angiogenic markers, and decreased expression of ChM-I. CONCLUSION: High expression of ChM-I was detected in articular cartilage of growing and normal adult joints, implicating its role in the maintenance of avascularity of intact articular cartilage. Expression of ChM-I decreased, while expression of VEGF and other pro-angiogenic factors increased, in OA cartilage. These findings suggest the loss of ChM-I from articular cartilage might be responsible in part for promoting blood vessel invasion into the cartilage during progression of OA. PMID- 14528520 TI - Measurement of articular cartilage surface irregularity in rat knee contracture. AB - OBJECTIVE: To design novel quantitative methods to evaluate the irregularity of articular cartilage surface; and to apply these methods for assessment of surface irregularity in a rat knee contracture model. METHODS: A total of 117 rat knees were either immobilized or sham-operated and harvested after 2, 4, 8, 16, or 32 weeks, and 11 knees were not operated. Standardized histologic sections were digitized and the contours of femoral and tibial cartilage surfaces were delineated. The rates of change in cartilage contour were calculated. Rate of change above a defined threshold constituted surface irregularity. RESULTS: In non-operated knees, cartilage surface irregularity in femur and tibia amounted to 3.1 +/- 0.5%. Immobilized knees showed significantly more irregularities than the sham-operated knees at all time points (2 weeks: 5.3 +/- 0.6% vs 3.1 +/- 0.4%; 4 weeks: 10.5 +/- 0.9% vs 4.4 +/- 0.9%; 8 weeks: 12.0 +/- 1.8% vs 4.9 +/- 0.2%; 16 weeks: 13.7 +/- 2.0% vs 4.9 +/- 0.4%; and 32 weeks: 13.8 +/- 1.4% vs 3.4 +/- 0.6%; all p < 0.05). No difference was observed between sham-operated and non operated knees. Increasing duration of immobilization in weeks (t) significantly correlated with more surface irregularity, described by the logarithmic formula: % irregularity = 6.6 + 2.1 ln (t), (F = 59.3, p < 0.001). This formula showed that irregularity progressed rapidly after immobilization and plateaued after 8 weeks. CONCLUSION: We designed methods to quantify cartilage surface irregularity and applied them to a contracture model. Cartilage surface irregularities appeared after 2 weeks of immobilization and progressed rapidly to plateau after 8 weeks. Combined with microscopic magnetic resonance imaging, this measurement of cartilage surface irregularity may constitute a sensitive tool to detect cartilage degeneration clinically. PMID- 14528521 TI - Rates of serious gastrointestinal events from low dose use of acetylsalicylic acid, acetaminophen, and ibuprofen in patients with osteoarthritis and rheumatoid arthritis. AB - OBJECTIVE: The frequency of serious gastrointestinal (GI) complications has been quantitated with chronic high doses of nonsteroidal antiinflammatory drugs (NSAID), but risk at lower dosages remains unknown. We examined the prevalence of serious GI events in patients taking aspirin (ASA), acetaminophen (APAP), or ibuprofen (IBU), focusing on low or intermittent use. METHODS: We studied 5692 patients with rheumatoid arthritis (RA) and 3124 patients with osteoarthritis (OA) from 12 databank centers, with 36,262 patient-years of observation, who had taken one of 3 study analgesics, and examined the frequency of serious GI events requiring hospitalization. RESULTS: Treatment groups were of similar ages and severity. As lower doses of study analgesics were taken, serious GI events tended to be less prevalent. In patients taking a study drug alone, without other analgesics or corticosteroids, only one event occurred in over 900 patient-years of exposure, roughly equivalent to background. Rates of GI events while taking APAP with other concurrent therapy or corticosteroids were higher (p < 0.05) than for the other 2 analgesics. In over-the-counter (OTC) doses, there were no significant differences in GI toxicity among analgesics. RA patients tended to have higher rates than OA patients. The rate of GI events was highly dependent on concurrent therapy, increasing 2 to 6-fold in patients taking other analgesics or corticosteroids. Propensity scores for serious GI events were similar across drugs. CONCLUSION: OTC use of ASA, IBU, or APAP carries little risk of serious GI toxicity for most persons. Most serious problems encountered were in higher-risk patients. Given the low rates of events, at low or intermittent dosage without concurrent treatment, these 3 analgesics cannot be distinguished from each other or from background rates of serious GI toxicity. PMID- 14528522 TI - The risk of Stevens-Johnson syndrome and toxic epidermal necrolysis associated with nonsteroidal antiinflammatory drugs: a multinational perspective. AB - OBJECTIVE: To quantify the risk of the severe cutaneous adverse reactions Stevens Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) associated with use of nonsteroidal antiinflammatory drugs (NSAID). METHODS: Three large data sources were analyzed: an international case-control study on severe cutaneous reactions (SCAR Study), a population based registry in Germany, and the US Food and Drug Administration (FDA) spontaneous reporting system. RESULTS: In the international case-control study, the oxicams were associated with the greatest increase in risk of SJS and TEN (relative risk 34, 95% confidence interval 11-105). When the risk for only recently initiated use was compared to that for longterm use of these agents (> 8 weeks), the relative risk of SJS and TEN associated with oxicams was significantly increased (p < 0.05). German data registry confirm these findings. The incidence of spontaneous US reports of SJS and TEN (per 1,000,000 visits with a prescription) for diflusinal, sulindac, oxaprozin, and etodolac were not significantly lower than that of piroxicam (p > 0.05, all comparisons). CONCLUSION: Although the absolute risks of SJS and TEN associated with NSAID use are low, these risks should be considered in monitoring patients who recently began therapy. In 3 independent databases, oxicams have higher risk of SJS and TEN than other NSAID widely used on the 2 continents. The FDA spontaneous reporting systems suggest some NSAID not widely used in Europe may have risks comparable to the oxicams. PMID- 14528523 TI - Predicting the risk of gastrointestinal bleeding due to nonsteroidal antiinflammatory drugs: NSAID electronic assessment of risk. AB - OBJECTIVE: To validate that, using patient demographics and other risk factors readily obtained from computerized databases, one can predict a priori the risk for developing a nonsteroidal antiinflammatory drug (NSAID) associated gastrointestinal (GI) bleed prior to exposing patients to therapy. METHODS: We conducted a retrospective cohort analysis using computer-stored information from a large group-model health maintenance organization. All patients who received one or more prescriptions for a single NSAID over a 9 month period were eligible. Historical and risk factor data was obtained for age, sex, prior GI bleeds, use of GI medications, prednisone use, and use of disease modifying antirheumatic drugs (DMARD). We tested a model (eSCORE) that is based on a previous risk stratification method. The primary outcome was a hospital admission for a GI bleed (GI event). RESULTS: A total of 303,211 NSAID patient-users met eligibility requirements. Serious GI events occurred in 302 patients, for a rate of 0.68% (0.68 events per 100 patient-years' exposure). All the risk factors except DMARD use were associated with a significant increase in the GI event rate. Higher eSCORE points were associated with increased GI event rates. CONCLUSION: The study supports the concept that the rate of GI events can be predicted by a defined set of easily assessed patient criteria using the eSCORE. Stratification of patients by risk score can guide the physician to appropriate therapeutic options, with the potential of protecting patients at greatest risk for a GI event. PMID- 14528524 TI - Prevalence of rheumatic symptoms, rheumatoid arthritis, ankylosing spondylitis, and gout in Shanghai, China: a COPCORD study. AB - OBJECTIVE: To carry out a cross-sectional survey on prevalence of musculoskeletal symptoms, rheumatoid arthritis (RA), ankylosing spondylitis (AS), and gout. METHODS: In Shanghai, 4 communities comprising 7603 inhabitants over 15 years of age in an urban population were randomly selected from 13 communities. Interviews were conducted from September 1997 to March 1998 by trained physicians using the COPCORD Core Questionnaire. Physical and radiographic examinations and serologic tests were carried out when required to classify categories of rheumatic diseases. The diagnoses of RA, systemic lupus erythematosus (SLE), and gout were based on American Rheumatism Association criteria. The diagnosis of AS strictly followed the modified New York criteria of 1984. Crude prevalence rates were standardized according to a standard Chinese population for age and sex structure. RESULTS: A total of 6584 adults (3394 women, 3190 men) were interviewed, and response rate was 86.6%. The age and sex standardized prevalence rate of rheumatic symptoms at any site amounted to 13.3% (95% CI 12.5-14.1%). Symptoms occurred more frequently in the following sites: knee 7.0% (95% CI 6.4 7.6%), lower back 5.6% (95% CI 5.0-6.2%), shoulder 4.7% (95% CI 4.2-5.2%), and neck 2.4% (95% CI 2.0-2.8%). Women complained of rheumatic symptoms more frequently than men. The standardized rates of RA, AS, gout, symptomatic knee osteoarthritis, and soft tissue rheumatism were 0.28% (95% CI 0.15-0.41%), 0.11% (95% CI 0.03-0.19%), 0.22% (95% CI 0.11-0.33%), 4.1% (95% CI 3.6-4.6%), and 3.4% (95% CI 3.0-3.8%), respectively. Two cases of SLE, one case of dermatomyositis, and one case of systemic sclerosis were found. CONCLUSION: Compared with rates in European and Western countries the prevalence rates of RA, AS, and gout are low in Shanghai, China, although the prevalence rates of rheumatic symptoms are high. PMID- 14528525 TI - Prevalence of the rheumatic diseases in urban Vietnam: a WHO-ILAR COPCORD study. AB - OBJECTIVE: To determine the prevalence rates of musculoskeletal disorders in an urban Vietnamese population. METHODS: The Community Oriented Program for Control of Rheumatic Disease (COPCORD) Stage I study was carried out in 16 groups in the Trung Liet Commune, Dong Da District, Hanoi City, Vietnam. Phase 1: the WHO ILAR COPCORD Core Questionnaire was applied by primary health care workers to 2119 urban subjects aged 16 years and over. Phase 2: 276 positive responders who had musculoskeletal complaints were interviewed by nurses and examined one week later. Phase 3: 261 positive responders in phase II were examined by 3 rheumatologists and 38% of these subjects required radiographic and blood tests to classify rheumatic disease categories. RESULTS: The response rates were 94.4%, 86.2%, and 94.6% in phases 1, 2, and 3, respectively. The prevalence of musculoskeletal pain was 14.9%. The most common musculoskeletal complaints were knee pain 18.2%, low back pain 11.2%, and soft tissue disorder 15.4%. Functional disability was reported in 6.04% of the survey population. The prevalence of rheumatic diseases was OA 4.1%, rheumatoid arthritis 0.28%, osteoporosis 0.47%, connective tissue disease 0.09%, and gout 0.14%. CONCLUSION: The prevalence of musculoskeletal pain in 2119 adults in an urban population in Vietnam was 14.5%, and osteoarthritis was the most commonly found arthritis. PMID- 14528526 TI - The efficacy of mindfulness meditation plus Qigong movement therapy in the treatment of fibromyalgia: a randomized controlled trial. AB - OBJECTIVE: To test the short and longterm benefits of an 8 week mind-body intervention that combined training in mindfulness meditation with Qigong movement therapy for individuals with fibromyalgia syndrome (FM). METHODS: A total of 128 individuals with FM were randomly assigned to the mind-body training program or an education support group that served as the control. Outcome measures were pain, disability (Fibromyalgia Impact Questionnaire), depression, myalgic score (number and severity of tender points), 6 minute walk time, and coping strategies, which were assessed at baseline and at 8, 16, and 24 weeks. RESULTS: Both groups registered statistically significant improvements across time for the Fibromyalgia Impact Questionnaire, Total Myalgic Score, Pain, and Depression, and no improvement in the number of feet traversed in the 6 minute walk. However, there was no difference in either the rate or magnitude of these changes between the mind-body training group and the education control group. Salutary changes occurring by the eighth week (which corresponded to the end of the mind-body and education control group sessions) were largely maintained by both groups throughout the 6 month followup period. CONCLUSION: While both groups showed improvement on a number of outcome variables, there was no evidence that the multimodal mind-body intervention for FM was superior to education and support as a treatment option. Additional randomized controlled trials are needed before interventions of this kind can be recommended for treatment of FM. PMID- 14528527 TI - Rupture of silicone gel breast implants and symptoms of pain and fatigue. AB - OBJECTIVE: To compare symptoms of women with silicone gel breast implants and women with chronic fatigue syndrome (CFS), and to study the effect of rupture of the silicone implant. METHODS: Five hundred readers of the Dutch silicone breast implant support group magazine were asked to respond if they had been informed by the surgeon about the silicone implant status at operation, and to answer questions about symptoms of CFS. Their complaints were compared with those of 100 female patients with CFS and 40 female controls. RESULTS: The questionnaires were returned by 319 women. Of these, 227 had symptoms of debilitating chronic fatigue. The patterns of symptoms differed from those in patients with CFS. An analysis of the relation between integrity of the implants and the symptoms could be carried out in 176 women, and 74% of these latter women reported ruptured implants. Significantly more women with ruptured implants than those with intact implants had debilitating chronic fatigue (75% vs 51%), postexertional malaise > 24 h (77% vs 51%), impaired short term memory (58% vs 38%), and multi-joint pain (77% vs 60%). CONCLUSION: Women with silicone breast implants often report severe pain and chronic fatigue. Rupture of the implant is associated with an increase in symptoms of pain and chronic fatigue. PMID- 14528528 TI - A comparison of the EuroQol-5D and the Health Utilities Index mark 3 in patients with rheumatic disease. AB - OBJECTIVE: To compare the performance of 2 commonly used utility-based health related quality of life (HRQoL) instruments [the EuroQol-5D (EQ-5D) and Health Utilities Index mark 3 (HUI3)] in patients with rheumatic disease. METHODS: Consecutive outpatients with rheumatic diseases were interviewed twice within 2 weeks using a standard questionnaire containing the EQ-5D, HUI3, and the Medical Outcome Study Short-Form 36 Health Survey (SF-36, used to categorize health status) and assessing clinical and demographic characteristics. EQ-5D and HUI3 utility scores were compared and their construct validity and test-retest reliability were examined by comparing these scores in groups differing in health status and using intraclass correlation coefficients (ICC), respectively. RESULTS: EQ-5D and HUI3 utility scores in 114 patients differentiated well between varying health states; e.g., patients with higher SF-36 vitality scores had better EQ-5D and HUI3 utility scores (mean: 0.79 for both instruments) than patients with lower vitality scores (mean: 0.68 and 0.69, respectively) (p < 0.01 for both comparisons). ICC values for the EQ-5D and HUI3 were 0.64 and 0.75, respectively (n = 90, median interval: 7 days). EQ-5D and HUI3 utility scores were similar (mean +/- SD: 0.75 +/- 0.21 vs 0.76 +/- 0.17, p = 0.647, paired t test) and showed moderate correlation (Spearman's r: 0.45, p < 0.001). Differences were present in patients' responses to these 2 instruments: e.g., 12 patients reporting no problems with mobility (EQ-5D item) reported different levels of disability with ambulation (HUI3 item). CONCLUSIONS: The EQ-5D and HUI3 performed equally well in measuring utility-based HRQoL in patients with rheumatic disease, although they measured slightly different, though related, dimensions of health. PMID- 14528529 TI - Incidence of juvenile idiopathic arthritis in the Nordic countries. A population based study with special reference to the validity of the ILAR and EULAR criteria. AB - OBJECTIVE: To find the incidence of juvenile arthritis according to the ILAR and EULAR criteria within defined areas in the Nordic countries, and to study the validity of the ILAR and EULAR criteria from this perspective. METHOD: A longitudinal, prospective, population based study with patients enrolled according to the ILAR and EULAR criteria. Twenty doctors in Iceland, Norway, Sweden, Denmark, and Finland collected data from the incidence cases within their catchment areas over a period of 1.5 years, beginning July 1, 1997. Clinical and serological data from the first year of the disease were collected. RESULTS: In the whole group of 315 patients, the incidence rate was 15 per 100,000 children/year (95% CI 13-17) according to the ILAR criteria, varying from 7 (1 13) in Iceland, 19 (7-31) and 23 (10-36) from 2 different regions in Norway, and 9 (5-12) and 16 (9-23) from 2 different areas in Denmark, to 15 (12-18) in Sweden and 21/100,000/year (15-26) in the Helsinki region in Finland. An early peak in distribution for age of onset was found in girls but not in boys. The number of antinuclear antibody (ANA) positive children in the whole group, made up of children who had undergone at least one analyzed ANA test, was 123/315 (39%). Girls were ANA positive in 83/197 (42%) and boys in 40/118 (34%). Uveitis developed in 27/315 (8.6%) children during the first 6 months of the disease. CONCLUSION: Incidence rates of juvenile arthritis for areas within the Nordic countries were in accord with previous data. The ILAR criteria present slightly higher incidence rates, with a shorter disease duration for inclusion, compared to the EULAR criteria. Patients in one subgroup in either of the criteria sets do not necessarily belong to the expected subgroup in the other set of criteria; e.g., for juvenile ankylosing spondylitis (EULAR) and enthesitis related arthritis (ILAR). Our epidemiological findings are a reminder to be aware of possible new subgroups in children with juvenile arthritis. PMID- 14528530 TI - Jaw claudication in primary amyloidosis: unusual presentation of a rare disease. AB - We describe two patients with temporal artery biopsy-proven amyloidosis presenting with symptoms of jaw claudication, visual disturbance, and proximal muscle stiffness suggestive of giant cell arteritis (GCA) and polymyalgia rheumatica. At the onset of disease, neither patient had other characteristic symptoms to suggest primary amyloid. We point out similarities between GCA and primary amyloid that can lead to confusion in diagnosis. PMID- 14528531 TI - Anti-tumor necrosis factor-alpha therapy-induced vasculitis: case series. AB - As experience with anti-tumor necrosis factor (TNF-alpha) therapy increases, there has been the expected emergence of reports on uncommon side effects. Large clinical trials identified the development of autoantibodies and postmarketing surveillance has identified problems including tuberculosis. There have been several case reports of drug-induced systemic lupus erythematosus. We describe eight patients with rheumatoid arthritis treated with anti-TNF therapies who developed presumed vasculitis, with different pathophysiologic causes. We discuss the literature and potential causal mechanisms, including disease activity, the role of autoantibodies, and shifts in T cell responses. PMID- 14528532 TI - Azithromycin-induced leukocytoclastic vasculitis. PMID- 14528533 TI - Human immunodeficiency virus-1 and rheumatoid arthritis: a puzzling association. PMID- 14528534 TI - Antibody response to influenza immunization in patients with systemic lupus erythematosus. PMID- 14528536 TI - Abstracts of the 5th Congress of Toxicology in Developing Countries. Guilin, China, November 10-13, 2003. PMID- 14528535 TI - Autonomic involvement in Sjogren's syndrome. PMID- 14528538 TI - Revisiting the approach to treatment of long-term illness: from the acute to the chronic state. A need for educational and managerial skills for long-term follow up. AB - The initial training of physicians and nurses is in the acute medical system, whether dealing with diagnosis or treatment of crises. This professional activity has gradually shaped the professional identity and is based on direct control, on avoiding risks and using therapeutic algorithms. When healthcare providers have to face chronic diseases and long-term follow-up strategies, this initial identity may often be counter-producing. This article describes the differences between the acute and chronic dimensions of diseases and treatments. Chronicity imposes on the healthcare provider a totally different way of functioning where he treats indirectly and should help the patient to manage the disease. Medical training has not put sufficient emphasis on the difference between those two approaches to disease. There is an urgent need for specific training in the strategies of management of long-term diseases. PMID- 14528539 TI - The Brief Medication Questionnaire: a tool for screening patient adherence and barriers to adherence. AB - Self-report tools for monitoring adherence can be useful in identifying patients who need assistance with their medications, assessing patient concerns, and evaluating new programs. The aim of this study is to test the validity of the Brief Medication Questionnaire (BMQ), a new self-report tool for screening adherence and barriers to adherence. The tool includes a 5-item Regimen Screen that asks patients how they took each medication in the past week, a 2-item Belief Screen that asks about drug effects and bothersome features, and a 2-item Recall Screen about potential difficulties remembering. Validity was assessed in 20 patients using the Medication Events Monitoring System (MEMS). Results varied by type of non-adherence, with the Regimen and Belief Screens having 80-100% sensitivity for "repeat" non-adherence and the Recall Screen having 90% sensitivity for "sporadic" non-adherence. The BMQ appears more sensitive than existing tools and may be useful in identifying and diagnosing adherence problems. PMID- 14528540 TI - The impact of tailored interventions on a community health center population. AB - We conducted a 4-year randomized study in a community health center that serves primarily low income Blacks in Durham, North Carolina. Patients (1318 at baseline) were assigned randomly to one of three study groups: provider prompting intervention alone, provider prompting and tailored print materials or the previous group and tailored telephone counseling. The purpose of the study was to determine whether increasingly intensive, tailored print and telephone interventions also were increasingly effective in promoting adherence to mammograms, Pap tests and overall cancer screening compliance. Thus, the combination of tailored print interventions (print and telephone) should have been more effective than the provider prompting intervention alone, or the print intervention and prompting combination. This is one of the few studies to examine a measure of overall cancer screening compliance and to assess the benefit of combinations of tailored interventions in promoting adherence to cancer screening. Patients gave extremely high ratings to the interventions. At the bivariate level, we found a significant effect of the most intensive group (provider prompting intervention, tailored print communications and tailored telephone counseling) on Pap test compliance (P = 0.05) and borderline significance at the multivariate level (P = 0.06) as well on overall screening compliance (P = 0.06). There was not a significant effect on mammography, probably because a majority of the patients were receiving regular mammograms. We also found some important subgroup differences. For example, a larger proportion of women reported Pap tests in the tailored print and counseling group when they believed the materials were 'meant for me.' These results show that a combination of tailored interventions may have potential for reaching the women who have too often been labeled the 'hard to reach.' PMID- 14528541 TI - Does arthritis influence perceived ability to fulfill a parenting role? Perceptions of mothers, fathers and grandparents. AB - The presence of a painful, disabling chronic disease may have implications for perceived ability to fulfill a parenting role. The purpose of this research was to examine the realities of parenting from the perspectives of mothers, fathers and grandparents with arthritis using a combination of methods: a cross sectional survey and in-depth focus group discussions. There was consensus that pain, fatigue and restricted physical functioning combined to interfere with the parenting role. Overall, approximately 35% of the sample had experienced difficulties attributed to arthritis. A gender difference emerged with women reporting more difficulties in relation to caring for babies and toddlers, whereas men reported more problems as children grew older. Key themes concerned: physical limitations; practical and caring issues; social factors; emotional response; hereditary risks and safety issues. Perceived inability to fulfill parenting roles resulted in feelings of frustration, guilt, anger and depression. A number of positive outcomes were mentioned including children's increased awareness of the needs of others. Limitations of the methodological approach adopted are discussed. PMID- 14528542 TI - Norms and prenorms on prenatal diagnosis: new ways to deal with morality in counseling. AB - This paper consists of two parts. In the first part, we report the results of an empirical study on Dutch women's decisions about prenatal diagnosis. We were interested in the norms these women used as guidance. One clear norm was found that was shared by everyone: everybody should decide for herself. Despite this clear norm, almost all women expressed difficulty with the choice, often regretted the fact that they had to make it and sometimes even expressed disgust about having to choose. These findings indicate that the choice is often experienced as a burden, which we think should be taken more seriously. In the second part of the paper, we present a way to do so. The core of our proposal is to pay more attention in counseling to the moral character of choices about prenatal diagnosis. We discuss difficulties and possibilities in that direction. PMID- 14528543 TI - Educating patient educators: enhancing instructional effectiveness in physical therapy for low back pain patients. AB - The objective of this research project was to study the effectiveness of a training program for the enhancement of patient education skills in physical therapy. In this paper the improvement of five of these skills is tested. These skills are aimed at a better monitoring of adherence problems during the treatment and at enhancing self-efficacy of the patient after treatment. In order to test the effectiveness of the program, complete treatments of 19 physiotherapists have been assessed before (1142 sessions, 130 patients) and after (775 sessions, 88 patients) the training program. Information on the instructions and solutions given to the patients was obtained with a registration form, completed after each session by the physiotherapist. The patient's perception of the effectiveness and feasibility of instructions was obtained from questionnaires, completed by the patient on three occasions. After the training only a minority of the trained skills appeared to be improved. All in all, the training program was not very effective. More effort is needed to develop training programs aimed at promoting patients' self-efficacy as well as measurement instruments to assess the effects of such programs. PMID- 14528544 TI - The effects of providing personalized dietary feedback. A semi-computerized approach. AB - There not only seems to be a trend for people to underestimate the dietary risks that they face, but it appears that this underestimation may be related to the difficulties they encounter when trying to assess their own dietary intake. A study (n = 118) examining the effects of providing people with information about their own dietary fat intake on their attitudes towards dietary change and their subsequent fat consumption is described. Participants in a group receiving feedback about their fat intake did not decrease their consumption of fat more than did those in a control group. No effects on subsequent fat consumption were observed in a group who had higher than average levels of fat consumption, higher perceived fat consumption than actual fat consumption and who received feedback information about their fat consumption. The results are discussed in relation to their implications for health promotion strategies that focus on the motivational effects of providing people with information about their fat consumption. PMID- 14528545 TI - Communication teaching and assessment in medical education: an international consensus statement. Netherlands Institute of Primary Health Care. AB - The importance of communication between doctors and patients has been well established, and there is growing acceptance of the need to teach and assess communication skills in medical schools. Faculty meeting at a consensus workshop during the International Conference on Teaching Communication in Medicine (Oxford, July 1996) generated a series of recommendations for developing and implementing teaching and assessment programmes. The points were refined in subsequent discussions with other interested groups, and endorsed in their current form by a workshop of teachers attending the Communication in Health Care Conference organized by NIVEL, the Netherlands Institute of Primary Health Care (Amsterdam, June 1998). While focused on medical schools, the eight recommendations highlighted in this consensus statement are also relevant to both graduate and continuing medical education programmes: (1) teaching and assessment should be based on a broad view of communication in medicine; (2) communication skills teaching and clinical teaching should be consistent and complementary; (3) teaching should define, and help students achieve, patient-centred communication tasks; (4) communication teaching and assessment should foster personal and professional growth; (5) there should be a planned and coherent framework for communication skills teaching; (6) students' ability to achieve communication tasks should be assessed directly; (7) communication skills teaching and assessment programmes should be evaluated; (8) faculty development should be supported and adequately resourced. PMID- 14528547 TI - Psychosocial factors in the etiology of breast cancer: review of a popular link. AB - Breast cancer is the most frequently occurring type of cancer in women in the western world. The etiology of a large proportion of breast cancers is still unexplained, and the possibility that psychosocial factors could play a role is not ruled out. Already in pre-Christian times, it was assumed that psychological factors might play a significant role in the development of breast cancer. However, studies have failed to produce conclusive results. There is still a lack of knowledge on the relationship between breast cancer development and psychosocial factors such as stressful life events, coping styles, depression, and the ability to express emotions. The results of this review show that there is not enough evidence that psychosocial factors like 'ways of coping' or 'non expression of negative emotions', play a significant role in the etiology of breast cancer. PMID- 14528548 TI - Psychosocial correlates of cancer relapse and survival: a literature review. AB - This article reviews literature on the psychosocial correlates of cancer relapse and survival from 1979 through 1995. The factors studied were structured according to a theoretical model of coping with cancer. Reviewed studies have shown that factors most frequently evaluated were depression, anxiety, hopelessness/helplessness, hostility, marital status and social involvement. Mainly inconsistent results were found. The strongest evidence for a relationship between psychosocial variables and prognosis was found for social involvement/social support; in 7 of 15 studies a positive relationship was demonstrated, while no negative associations were found. Coping styles e.g., fighting spirit and stoic acceptance, and severe/stressful life events were found to have no conclusive influence on the length of survival. Important determinants of the coping model, such as uncertainty and information given by the specialist were not studied as possible predictors of survival and/or relapse free period. Among the factors that showed no correlation at all was multidimensional health locus of control. For the inconsistent findings, a considerable number of methodologic shortcomings with respect to study design, sample size, measure and statistical analysis are enumerated. PMID- 14528549 TI - Evaluation of an educational programme for the early detection of cancer. AB - Avoiding patient's and doctor's delay is important for the detection of cancer. In order to study the possibilities for shortening the delay, without causing anxiety, an educational programme for early detection of cancer (EPEDC) was worked out, aimed to be evaluated at the community level. A community with 77,100 inhabitants, was informed about cancer symptoms in a letter. Participants who observed the cancer symptoms, described in the letter, were invited to visit the health centres, where they were interviewed and examined according to a specially designed schedule. Guidelines for taking care of these participants were also worked out. Fifteen previously unknown cancers were detected. By means of a telephone interview and a questionnaire the reactions to the EPEDC were studied. The results indicate that it is possible to inform and educate the population about cancer symptoms without causing anxiety on condition that there is an organisation which can be contacted without delay by subjects with potential cancer symptoms. PMID- 14528550 TI - South Asian womens' views on the causes of breast cancer: images and explanations. AB - Using ethnoscience methods, interviews with 50 South Asian women living in Western Canada were conducted and analyzed to explore explanations and images of breast cancer. Embedded in the women's stories of breast cancer were distinctive, often vivid and fear-provoking images of abnormal growth. Explanations about the causes of breast cancer involved five domains of belief. The first domain was of a physical nature and centred on damage to the breast. A second domain of explanations, 'can catch it,' focused on the way this disease could be spread to others. Other women attributed breast cancer to the ways women could 'bring it upon yourself,' often linking a negative lifestyle with the development of cancer. Many women attributed cancer to being 'in the hands of others,' explaining the cancer was caused by careless words, curses or divine power. Finally, breast cancer was seen as something that could be passed down in the family. The taxonomy developed in this study provides a useful framework for understanding the explanations that might underlie women's health-seeking behaviours and for developing culturally suitable counseling strategies. PMID- 14528551 TI - Information and patient participation in screening for prostate cancer. AB - The purpose of this study was to determine if providing men with information about screening for prostate cancer would enable them to assume a more active role in decision making with their family physician, and lower levels of anxiety and decisional conflict. Men were recruited from one family medical clinic in Winnipeg, Manitoba. One hundred men scheduled for a periodic health examination (PHE) were randomly assigned to receive verbal and written information either prior to the PHE, or following the second interview. Men completed measures of preferred decisional role and anxiety prior to the PHE; and assumed decisional role, decisional conflict, and anxiety post PHE. Results demonstrated that men who received the information prior to the PHE assumed a significantly more active role in making a screening decision, and had lower levels of decisional conflict post PHE. The two groups did not differ with regard to levels of state anxiety. Providing men with information enables them to make informed screening decisions with their family physicians. PMID- 14528552 TI - Chronic and acute stress as predictors of relapse in primary breast cancer patients. AB - In this paper we present data on predictors of early relapse (at three and a half years post-surgery) in 44 patients with primary breast carcinoma. We expected that the degree to which receiving the diagnosis of malignancy would act as an acute stressor would allow a prediction of early relapse. Acute stress is measured by means of changes in psychological, neurochemical and immunological indicators of stress before and after the communication of the diagnosis. The results indicate that the assumption does not hold. Chronic stress, however, appears to be a strong predictor of early relapse. The measure of chronic stress is based on a self-report questionnaire of health complaints in the year before the diagnosis of breast cancer. PMID- 14528553 TI - Preferences for different sources of information about cancer. AB - A wide range of cancer information resources is available to patients and their relatives, who are increasingly acting as independent learners. In order to examine preferences for different types of information and to establish the appropriateness of providing a Web-based cancer information resource (CancerHelp UK) a questionnaire survey was undertaken. Of 258 questionnaires distributed to patients attending for oncology outpatient or general practice appointments, 205 (80%) were suitable for analysis. 74% (95% confidence interval 68% to 80%) of participants wanted as much information as possible. All participants cited people most frequently as a future source of cancer information. All participants frequently cited television as a source of both general and cancer-specific information. Despite a lack of computer experience patients were significantly more likely than other groups to say that they would like to use the Web-based resource. It is suggested that such resources are likely to be increasingly well used, particularly by cancer patients, given the growth in health information on the Web and developments in delivery of digital information into the home via television. PMID- 14528555 TI - Coping with cancer. A personal odyssey. AB - Building on personal experience, the psychological impact of cancer is reviewed. The subspecialty of psycho-oncology is a recent development within oncology, reflecting the increased interest in behavioral, psychological, and social factors in cancer prevention, treatment, and in improving the quality of life of patients with cancer at all stages through counseling and educational programs. Early identification of patients who are coping poorly is important for compliance with treatment and control of distress. It is equally important to recognize and diagnose common psychological disorders, primarily anxiety and depressive symptoms, which can occur. Referral for counseling to a mental health professional familiar with the care of patients with cancer may be indicated. My successful confrontation with cancer led to emotional growth. For those who cannot be sanguine about their experience, there may still be ways to reduce its, at times, devastating psychological impact. Hopefully, the days in which cancer patients had to suffer alone, in secrecy, and in fearful silence, are disappearing. PMID- 14528554 TI - Offering a choice between two adjuvant chemotherapy regimens: a pilot study to develop a decision aid for women with breast cancer. AB - BACKGROUND: The primary objective of this study was to develop a decision aid which would encourage and assist patients to become involved in treatment decision making, and help clinicians to objectively educate patients about the benefits and risks of adjuvant chemotherapy for breast cancer. A secondary objective was to investigate the factors influencing this treatment decision making process for women when choosing between adriamycin and cyclophosphamide (AC) versus cyclophosphamide, methotrexate and 5-fluorouracil (CMF) chemotherapy. METHODS: An educational visual instrument called a Decision Board was developed consisting of written and graphical material. The Decision Board displays general information about chemotherapy and detailed information about each chemotherapy regimen, including the schedule and side effects, and was presented to patients with a scripted standardized oral explanation. The instrument was evaluated in 46 premenopausal women newly diagnosed with node-positive breast cancer. Following presentation of the board, the patients were given a take home version to review and asked to return 1-2 weeks later with a decision. During the second visit each patient was asked to complete a questionnaire regarding demographics, learning and comprehension, treatment preference, and factors influencing their decision. RESULTS: Recall of information was acceptable (> or = 80%). The Decision Board was found helpful by all, but the level of difficulty with decision making was variable. Out of 46 women, 23 women chose AC, 21 chose CMF, and two chose no treatment. The major factors affecting treatment preference were related to the impact on quality of life, the length of therapy, and the side effects, in particular, vomiting and alopecia. CONCLUSIONS: The Decision Board appears to be a valuable educational tool that enables patients to become well-informed and directly involved in their treatment decisions. PMID- 14528556 TI - HIV/AIDS: new educational and counseling strategies. PMID- 14528557 TI - Assessing the application of HIV and AIDS related education and counselling on the Internet. AB - This study reviewed Internet technological capabilities for counselling and several appropriate counselling models and assessed the application of HIV/AIDS related counselling on the Internet. Sixteen health professionals in HIV/AIDS related face-to-face counselling positions were interviewed: their HIV/AIDS service background was limited to Texas-certified HIV pre/post test counsellors. Texas-licensed counsellors in HIV/AIDS service field and HIV/AIDS case managers and social workers. Duration of interviews ranged from 30 min to 60 min and were recorded on audio cassette for review and analysis. Responses were generated using an editing style of the long-interview process. Edited responses were then analyzed for content and themes. Four major themes evolved from interview responses: counsellor-client relationship, target population, ethics and operation. Major concerns included the lack of visual and verbal cues during interaction, problems of accessibility by the neediest, confidentiality, impersonal experience and increased client separation/isolation. Greater benefits may be attained by targeting the younger segment of the population and other health professionals. A majority of respondents indicated support for additional development of Internet-based HIV/AIDS related counselling. PMID- 14528558 TI - Audio-taping the oncology treatment consultation: a literature review. AB - An increased amount of research has been conducted to evaluate interventions for improving the quality of communication between cancer patients and health care providers. One of these interventions involves providing patients with audiotapes of their consultations with oncologists. Given that effective patient-physician communication has been linked to beneficial health outcomes, an examination of studies that have evaluated the effects of audiotape provision appears warranted. This article provides a critical review of this literature. The audiotape intervention has been examined in uncontrolled studies and randomized trials, and the primary outcome variables have included psychological well-being, information recall, and patient satisfaction. The empirical literature is unclear as to the efficacy of providing patients with taped recordings of cancer consultations. Overall, the findings suggest that the majority of patients benefit from receiving the audiotape, but the utility of this intervention in improving patient-physician communication requires further examination. Replication studies and well-controlled experimental designs applied to a variety of health care providers in diverse oncology settings are needed to confirm the validity of the empirical findings to date, and to facilitate further development of interventions aimed at enhancing patient-physician communication. PMID- 14528559 TI - Targeted prevention for people with HIV/AIDS: feasible and desirable? AB - People with HIV/AIDS are rarely chosen as a target group for prevention activities. In this paper we look at empirical and theoretical evidence for the feasibility and desirability of directing preventive interventions at HIV positives. Research data on the behaviour and motivation of HIV-positives suggests that the differences between HIV-positives and HIV-negatives and those who are unaware of their HIV-status are not large. However, specific determinants of behaviour, such as responsibility for others or the risk of superinfection, have seldom been measured. Effective interventions targeting at HIV-positives and focussing on prevention are lacking. Fear of increased stigmatization has been used as an argument against focussing prevention activities at HIV-positives. Theoretically that argument is probably not correct: positive coping with HIV may invite positive reactions. The conclusion is that HIV-positives should be chosen as a special target group for additional planned preventive interventions. Because people need to be aware of their HIV status, testing and treatment sites are adequate settings. Effective interventions should be developed on the basis of theory and evidence about the specific determinants of risk behaviour of HIV positives: protecting oneself for superinfection and protecting one's partner. PMID- 14528560 TI - Family-oriented care during pediatric inter-hospital transport. AB - We evaluated the family-oriented care and counseling provided by the BC Children's Hospital Transport Team paramedies. One hundred families were asked to rate: (1) how they would like paramedies to communicate with them under ideal conditions; and (2) what they actually experienced during their child's transport. There were no significant differences in parents' preferences under ideal circumstances and what they actually experienced in five of nine behavioural areas studied. Nineteen paramedies also rated their use of elements of family-oriented communication and the strategies they would recommend other teams use when interacting with families. The elements that paramedies reported using most frequently were rated highly by parents and produced positive feelings in the majority of families. The study indicates that effective counseling can be achieved in 'critical', time-limited situations but improvements can be made; and it supports the value of appropriate selection and training of transport team personnel in family-oriented critical care. PMID- 14528561 TI - A survey of elderly diabetic patients attending a community clinic in Hong Kong. AB - A cross-sectional survey was conducted on Chinese diabetic patients attending a community clinic in Hong Kong. A questionnaire, physical examination and blood tests were administered. Of 155 adults with diabetes mellitus recruited, the mean age was 63 years (SD 10.3), 77.4% were female and 41.9% illiterate. Half the patients achieved adequate knowledge scores. Knowledge was found to be associated with education level (OR 4.3, 95% CI 1.8, 10.0), longer disease duration (OR 3.2, 95% CI 1.4, 7.4) and complications (OR 0.3, 95% CI 0.1, 0.8). No correlation was found between knowledge and attitude or practice. Attitude was found to be associated with practice (OR 1.2, 95% CI 1.0, 1.3) and male gender (OR 2.9, 95% CI 1.1, 7.5); practice was associated with hypertension (0.5, 95% CI 0.2, 0.9). Older less educated Chinese diabetic patients can be educated about diabetes management, but knowledge does not always translate into practice. PMID- 14528562 TI - AIDS knowledge and attitudes of pupils attending urban high schools in Israel. AB - The present study compared AIDS knowledge, attitude and sources of information of 1724 students in three different high schools (academic, vocational and religious) in Holon. Forty-two percent of the students are anxious concerning the possibility of AIDS contamination; they expressed a willingness to be tested for HIV and AIDS. Pupils attending the academic school proved to have the highest knowledge concerning AIDS topics; those in the vocational school placed second; and those in the religious school came last. The knowledge increased with age (p < 0.001). Results indicated that misconceptions were still found in a second decade of that epidemic concerning the following areas: etiology, ways of exposure; symptomatology; and prognosis. Their intolerant attitudes reflect social anxiety and vulnerability to AIDS. The most liberal and comprehensive attitudes toward AIDS patients were presented by pupils attending the academic school and after that came pupils from the vocational school, while pupils in the religious school presented the most conservative and intolerant attitudes. The sources of pupils' information primarily included: the media--TV (92.3%): newspapers (87.6%); and school (66.1%) occupied the third place. It is remarkable that physicians and nurses at school and outside clinics were placed in the seventh (and last) place, as only 25.3% mentioned them at all. Only one third of the pupils agreed to participate as volunteers in medical and rehabilitation centers which help AIDS patients. PMID- 14528563 TI - What are the eating cognitions of children whose chronic diseases do and do not require attention to diet? AB - Nutrition therapy is essential to the management of several chronic diseases affecting adolescents. Unfortunately, dietary impositions may evoke pathological eating-related cognitions. This pilot study examined eating- and weight-related cognitions of 55 adolescents attending a summer camp for youngsters with a heterogeneous variety of chronic diseases. Campers completed the Eating Cognitions Questionnaire and the Situational Obstacles to Dietary Adherence questionnaire. Campers whose medical regimens included special dietary recommendations (n = 24) were expected to have more cognitions associated with eating disorders than those without dietary restrictions (n = 31). Girls were significantly more concerned than boys about weight and restrictive dietary rules. Cognitions of youngsters with special dietary recommendations showed greater flexibility, less eating pathology and less weight concern than campers without recommended diets. Contrary to previous reports, dietary restrictions were associated with more appropriate eating cognitions, suggesting that adolescents with chronic illnesses requiring nutrition therapy may not necessarily bear greater risk of eating disorders. PMID- 14528564 TI - Adherence to HAART: impressions from the 12th world AIDS conference, Geneva, June 28-July 3, 1998. PMID- 14528566 TI - Parenting adolescents with ADHD: evaluation of a psychoeducation group. AB - ADHD and disruptive behaviour disorders are the most common reasons for referrals of adolescents to children's mental health services. Psychosocial treatments for parents and adolescents, though routinely recommended, have not been well evaluated. This paper reports an evaluation of a clinic based, parent skills training and education group for parents of adolescents with ADHD. The parent education group is a manual based program which provides information about ADHD and training in problem solving and parenting skills. Using a pre-post design, we collected evaluation data from 163 parents of 103 adolescents before and after participation in the group. The outcome measures were two parent self-report measures, the conflict behaviour questionnaire, the issues checklist and an investigator-designed evaluation questionnaire. There was a statistically significant decrease in frequency and intensity of self-reported parent adolescent conflict and in parent-reported problem behaviour. Parent reports of problem behaviour were not related to kin relationship or child age. There was a trend for parents of daughters to report more problem behaviours and more conflict in the parent-adolescent relationship. Participants reported positive effects on their parenting skills and confidence. They tended to evaluate the program as very helpful. PMID- 14528567 TI - Avoiding unwanted pregnancy--the role of communication, information and knowledge in the use of contraception among young Norwegian women. AB - Despite relatively easy access to contraceptives, a liberal attitude to pregnancies out of wedlock and a widespread family planning education in schools, too a high number of adolescent women in Oslo, the capital of Norway, become pregnant and resort to abortions. The aim of this study was to identify some new entry points to sexuality, contraceptive, abortion and post-abortion counselling. Using a qualitative approach, we interviewed 102 young women in Oslo who were seeking either an abortion or contraceptives. The study demonstrated that contraceptive awareness is good, but that there are different levels of consistency in contraceptive use among women, and that even women with few partners and fairly good contraceptive compliance sometimes experience unplanned pregnancy. The study reviews some issues of importance where communication with young women could be improved. These issues include better formal information about OCs, increased information on emergency contraception, better condom promotion, and an attempt to involve better informed adults, including paramedical professional counselling. In addition, contraceptive prescriptions should accord to the type of behaviour and the relationships the young women have. PMID- 14528568 TI - Change in self-efficacy during cardiac rehabilitation and the role of perceived overprotectiveness. AB - Self-efficacy (SE) is an important outcome following cardiac rehabilitation (CR) when claiming benefits to patients and improving existing programs. This study evaluated change in SE during 6 weeks of multidisciplinary CR with either a high or low-frequency exercise training program. The role of overprotectiveness of the spouse, as it potentially counteracts improvement in SE, was examined. Coronary patients (n = 114) were randomized into both types of program. Overprotectiveness (patient perception) was assessed prior to rehabilitation. Self-efficacy in the domains controlling symptoms (SE-CS) and maintaining function (SE-MF), were assessed prior to and immediately after rehabilitation. Three findings pertain to program improvement: (1) As predicted, the low-frequency program enhanced SE-CS more than the high-frequency program, suggesting that experiencing success in daily activities and active engagement of the patient seem more decisive factors in improving SE than the frequency of exercise. (2) Changes in SE in both programs fell short of clinical meaning, suggesting the need to use self-efficacy theory more vigorously. (3) Overprotectiveness significantly predicted adverse change in SE in the high-frequency program, suggesting the need to include counseling for cardiac couples in CR when applicable. PMID- 14528569 TI - Development of a brief test to measure functional health literacy. AB - We describe the development of an abbreviated version of the Test of Functional Health Literacy in Adults (TOFHLA) to measure patients' ability to read and understand health-related materials. The TOFHLA was reduced from 17 Numeracy items and 3 prose passages to 4 Numeracy items and 2 prose passages (S-TOFHLA). The maximum time for administration was reduced from 22 minutes to 12. In a group of 211 patients given the S-TOFHLA, Cronbach's alpha was 0.68 for the 4 Numeracy items and 0.97 for the 36 items in the 2 prose passages. The correlation (Spearman) between the S-TOFHLA and the Rapid Estimate of Adult Literacy in Medicine (REALM) was 0.80, although there were important disagreements between the two tests. The S-TOFHLA is a practical measure of functional health literacy with good reliability and validity that can be used by health educators to identify individuals who require special assistance to achieve learning goals. PMID- 14528570 TI - The attitude of young adults with chronic disease or handicaps towards enforced treatment and euthanasia. AB - One hundred fifty-nine young adults in the 17-18 year old age group suffering from chronic disease or handicap participated in this study which examined their attitudes towards enforced treatment and euthanasia in terminal patients. The comparison group included 120 healthy high school pupils of the same age group. A total of 42.6% of the chronic patients thought that enforced treatment was justified even if the patient didn't understand its importance and didn't want treatment (in comparison with 23.3% of the healthy pupils who felt the same way). Of the chronic patients, 54.4% thought that euthanasia was justified in consenting terminal patients (in comparison with 74.2% of the healthy pupils who felt the same way; P < 0.01). This may be explained by the feeling of total dependency of chronic patients upon medication and treatment. On the other hand, they may be opposed to euthanasia because of their own personal hope that a cure would be found for their severe and chronic condition. Both groups studied believed that physicians should always consider the subjective suffering of the patient and his family, as well as the short and long term prognosis when deciding about therapy. PMID- 14528571 TI - Perceptions of health care professionals and patients about the risk of HIV transmission through oral sex: a qualitative study. AB - We explored health care professionals' beliefs and methods for counseling patients about risks for transmitting human immunodeficiency virus (HIV) through oral sex and HIV-seropositive patients' beliefs and practices regarding oral sex behavior. Health care professionals used divergent counseling strategies, avoided specific recommendations, and expressed ambivalence about recommending barrier protection for oral sex. Patients expressed differing beliefs about oral sex risk; the majority have engaged in unprotected oral sex since diagnosis. Few professionals or patients mentioned oral sex risk for other sexually transmitted diseases (STDs), which can be cofactors for HIV. Although scientific evidence indicates a potential risk for transmitting HIV via oral sex, and patients in our sample want more information about this risk, the health care professionals we interviewed have adopted a largely noncommittal approach to communicating potential risk to patients. Health care professionals should consider discussing more specifically with patients the oral sex risk for transmission of HIV and other sexually transmitted diseases. PMID- 14528572 TI - A new approach for patient education: beyond constructivism. AB - Over the last ten years, research developed in science education, especially at the University of Geneva, has demonstrated that education must be centered on interactions between the learner's conceptions and a systemic educational environment. Transposed to a medical knowledge, this new model, originally conceived by the authors and called the 'allosteric learning model', offers new ways of considering patient education. After criticizing the 'grand theories' of learning, mainly the latest ones called constructivist models, the authors suggest a new set of micromodels designed to explain thoroughly the functioning of the patient's thought process (questions, frame of references, semantic network,...) and his understanding of medical information on his own disease. For health care providers, these models also offer a series of new pedagogical approaches both efficient and original to regulate the act of education based on confrontation, mobilisation, integration, etc. PMID- 14528574 TI - [Nursing becomes a profession]. PMID- 14528573 TI - Encouraging out-patients to make the most of their first hospital appointment: to what extent can a written prompt help patients get the information they want? AB - A randomised controlled study in which a written prompt was sent to new patients to help them make the most of their consultation was conducted amongst patients referred to the dermatology, gynaecology and orthopaedic out-patients clinics at the Royal Free Hospital, London. The impact of the help card on patients' expectations, preparation for and experience of their out-patient consultation are discussed. A help card and letter were sent to a random sample of patients before their appointment to encourage them to prepare and prioritize questions to ask the doctor at the consultation. After their consultation, patients were sent a postal questionnaire to complete at home. Analysis of the questionnaires provided quantitative and qualitative data about patients' information requirements and whether they were fulfilled. The results highlight the difficulties out-patients have in asking questions and discussing topics fully at their initial consultation, even when they have thought of questions in advance. Half the patients who were sent a help card said they got more out of their consultation as a result, yet few statistically significant differences between the help card group and the other patients were found. PMID- 14528575 TI - [Up and down--the experience of women with the rheumatic illness fibromyalgia]. AB - Chronic pain, like that inflicted by fibromyalgia, is not much known, unpredictable and can emerge everywhere in the body. Yet 2 to 4% of the population suffer from this disease, of which 73 to 88% are women. In fact fibromyalgia is more wide spread than the more known rheumatoid arthritis. This research work presents the experience of women in their struggle against this incurable disease and the progress made in the fight against it. The Chronic Illness Trajectory by Corbin and Strauss (1992) is used as a theoretical framework. Six women, over fifty years of age and suffering from fibromyalgia for over ten years, were interviewed. The interviews were audiotaped, transcribed verbatim and analysed using the grounded theory methodology. The instability during disease development, referred to in this paper as "up and down", is an important phenomenon. Along with the pain and "defencelessness" goes the feeling of "not being taken seriously". Results have proven that an interdisciplinary approach is the best way to help those that cope with fibromyalgia. PMID- 14528576 TI - [Nursing without and with essential oils: a controlled study of patients in an acute rheumatologic department]. AB - Before introducing essential oils in nursing in a whole clinic, a controlled pilot study was performed. Within the study frame, forty voluntary patients of a rheumatology ward in the City Hospital Triemli (Zurich, Switzerland) had each an indicated nursing intervention without and with essential oils. The main criteria of the nursing activities were physical contact between patient and nurse; no medication besides the prescribed ones, same procedure with and without essential oils. Data recorded immediately before and 30 to 45 minutes after nursing comprised the general well-being of patients in their own and their nurse's ratings, the patient's rating of his/her state with respect to all study nursing indications, and the nurse's self-rating with respect to her mental well-being. At discharge patients commented on their in-patient stay as well as on nursing with essential oils. The general well-being of study participants was comparable before either type of nursing intervention with the exception of the aspect of well-being due to which the intervention was done: ratings of these intervention specific aspects were much worse in relation to nursing with than without essential oils. However, specific well-being significantly improved only due to nursing with but not without essential oils. The larger the intervention-specific effect of nursing with essential oils, the significantly more positive were the comments on nursing with essential oils at the patients' discharge. INTERPRETATION: Nursing with essential oils has specific effects, but does not improve the general well-being. Nursing with essential oils is more efficacious than when oils are not applied. PMID- 14528577 TI - [Effects of kinesthetic mobilization in comparison with standard mobilization on respiratory function after coronary artery bypass and other relevant factors]. AB - Presently no relevant nursing data exist in Germany on the effect of mobilization according to kinaesthetic principles on the physiological system, on locomotion and the sensory system. The effect of two different versions of mobilization was measured in a unicentric, open, randomized study with two parallel groups from December 1999 to October 2000 in the intermediate ICU for patients after heart surgery at Ulm University Hospital. The aim was, to show superiority of kinaesthetic mobilization, compared to empirically developed standard mobilization on patients after aortal coronary bypass surgery, with reference to respiratory function. The two protocols of mobilization were carefully coordinated with precise plans of action and timing. A total of 104 subjects were randomized to the two groups. In the kinaesthetic mobilization group (n = 52) median age was 67.5 years, ten (19.2%) were female and 42 (80.8%) were male. In the standard mobilization group (n = 52) median age was 69 years, eight (15.4%) were female and 44 (84.6%) were male. The main outcome variable was the respiratory minute volume 30 minutes after the second mobilization minus the respiratory minute volume prior to the second mobilization. The median change in respiratory minute volume was 0.4 ltr/min. (-5.1 to 3) in the group with kinaesthetic mobilization and median 0.3 ltr/min (-6.0 to +9.1) in the standard mobilization group. Superiority of kinaesthetic mobilization compared to standard mobilization could not be proven (p = 0.38). Also in the secondary outcome variables the kinaesthetic mobilization showed no superiority over standard mobilization. The respiratory minute volume data within points of measure show that subjects in the kinaesthetic group have less extreme results (> 12 ltr.). In the area < 5 ltr. there is a similar tendency. This may point out that regulation between movement of body, respiration and circulatory system occurs more continuous in kinaesthetic mobilization than the standard mobilization. A similar tendency was seen in the secondary outcomes variables. Subjectively nurses noted that kinaesthetic mobilization was less strenuous than standard mobilization, even though subjects in the kinaesthetic group showed more mobility restriction (37%) than in the standard group (15%). From before to after the first mobilization the percentage of subjects needing less pain medication was higher in the kinaesthetic group (35%), compared to the standard group (19%). There were no serious adverse events in either group. The concept of kinaesthetic mobilization showed no negative effect on subjects compared to empirically developed standard mobilization. There was no diagnosis of pneumonia and none of instabile sternum on any of the subjects in this study. Also, there was no negative effect on the sternum due to the light support arms gave to sitting up and rotating movements in the area of thorax and pelvis on subjects in the kinaesthetic group. The subjects early trust in their own varied movement and the possible effects of kinaesthetic mobilization in later phases of recuperation was not explored in this study. PMID- 14528578 TI - [Voluntary helpers in hourly home care of patients with dementia. Intermediate term results of a prospective study and results of a retrospective survey]. AB - Care of home-based dementia patients by trained part-time volunteer "helpers" is seen as a future-oriented, easily accessible and financially affordable means of relieving the burden on family carers. Qualitative interviews with 30 former or currently active helpers were conducted, and information gathered about their past experience. The additional, prospective part of the study, which is as yet uncompleted, has so far provided data on 33 individuals with regard to their motivation in wishing to become involved in this type of voluntary relief work. The most common characteristics of this group are previous experience in the care of old and sick people, either as private citizens or in a work situation, and previous periods engaged in voluntary work. Many volunteers had had what they perceived as unpleasant experiences when caring for dementia patients. Such experiences do not lead to the abandoning of the voluntary activity so long as an experienced professional is readily available to give support and advice when there are problems. If the kind of voluntary relief work described here is to succeed in its aims then, in addition to the professional support, volunteers will require thorough preparation for their task. PMID- 14528579 TI - [Effects of health policy mandates on ambulatory care in Germany]. AB - For the last few years health politicians in Germany have passed a number of health care reform plans which have the aim to increase efficiency, effectiveness, transparency and quality in the area of health care services in order to reduce the costs of health care supply. A strategy is a shift from hospital to home. Recently passed health care measures follow this paradigm. In politics and sciences the consequences of these measures are mainly discussed for the doctors. On the basis of an analysis of Anglo-Saxon literature this article discusses the effects of health political measures on home care and asks which are the chances and possible developments for home care. The results show that home care can benefit from the developments in health care. Home care can take a central function and part in the process of change in the German health care system. Home care can take on tasks in the area of coordination of home health care and cooperation of multidisciplinary teams. Home health care must develop into a modern service profession, which takes an active part in the process of reorganisation of the German health care system. PMID- 14528580 TI - [Evidenced-based nursing: knowledge, performance and learning in nursing care]. AB - As part of the nursing process, nurses choose an academically secured intervention in response to a patient's problem, implement it and evaluate it. "Evidence based" describes a learning and working method which has the advantage of being applied in real, professional problem situations. Not only the learner but also the professional nurse can improve their competencies for professional acting. Following the problem- and motivation orientated approach of Roth (1983), the implicit learning steps of this method can be illustrated. The implementation of evidence-based nursing can raise high expectations. However, there are also critical aspects connected to it that need further exploration. PMID- 14528581 TI - Patient care through pregnancy termination. PMID- 14528583 TI - Exploring infertility & endometriosis. PMID- 14528582 TI - AWHONN then & now. PMID- 14528584 TI - Oral insulin doesn't prevent type 1 diabetes. PMID- 14528585 TI - U.S. birth rate hits record low. PMID- 14528586 TI - Donor immune cells attack metastatic breast cancer. PMID- 14528587 TI - Nurses in Washington D.C. PMID- 14528588 TI - Keeping up with the Women's Health Initiative. PMID- 14528589 TI - Irritable bowel syndrome in women. Exploring diagnoses & treatment options. PMID- 14528590 TI - Prescribing oral contraceptives. Focusing on each woman as an individual case. PMID- 14528591 TI - Ka'nistenhsera. Teiakotihsnie's. A Native community rekindles the tradition of breastfeeding. PMID- 14528592 TI - Violence against pregnant women. These two screening tools may prove valuable in identifying women at risk. PMID- 14528593 TI - Patient page. Preventing premature birth. PMID- 14528594 TI - Helping pregnant women kick the habit. PMID- 14528595 TI - How a hospital room becomes a window of opportunity. PMID- 14528596 TI - A pilot study of the elder abuse questionnaire. AB - This descriptive study's purpose was to pilot a questionnaire with sub-scales in a population of 40 family members listed as power of attorney (POA) for residents and 40 employees of a long term care facility in rural southeastern Kansas. The non-random convenience sample was 49 participants resulting in a 61.25% participation rate. The framework for the study was Betty Neuman Systems Model. Neuman's variables are related to the physiological, psychological, socio cultural, developmental, and spiritual stages of the multidimensional person (Neuman, 1982). The 25 items on the Elder Abuse Questionnaire (EAQ) were derived from these variables. PMID- 14528597 TI - Promoting pain management in Kansas: a collaborative effort. PMID- 14528598 TI - Code of ethics for nurses with interpretive statements. PMID- 14528599 TI - The testosterone connection. PMID- 14528600 TI - [The chemo-protector effects of tea and its components]. AB - Tea has been consumed worldwide since ancient times to maintain and improve health. Its main active components are a type of polyphenols known as flavonoids, which include catechins and theaflavins. Several epidemiological studies suggest that the consumption of green tea could prevent cancer development in humans. Likewise, animal studies have shown that green tea consumption may inhibit the development of prostate and breast cancer. It has been shown that, through several mechanisms, tea polyphenols present antioxidant and anticarcinogenic activities, thus affording several health benefits. It is important to better characterize tea components, to study their bio-availability and bio transformation in vivo and to conduct clinical studies of its main active compounds. PMID- 14528601 TI - [Iron metabolism: current concepts of an essential micronutrient]. AB - Iron is an essential micronutrient involved in multiple biochemical and physiological process. In this review we discuss the most relevant aspect of its metabolism in order to reach a better comprehension of the relevant roll that this micronutrient plays in human health. PMID- 14528602 TI - [Dietary evaluation of pregnant adolescents during first, second and third trimester]. AB - In order to identify risk of inadequate intake of calories and nutrients during pregnancy, 75 pregnant adolescents (14 to 18 years old) apparently healthy, from low socioeconomic level, in their first prenatal control (12 weeks) were studied by two 24 hour recalls and a food frequency questionnaire at each trimester. Nutritional status was assessed by body mass index (pregestation weight/height) and classified according to American Medicine Institute reference. Paired t-test, frequency distribution and ANOVA were used for statistical analysis. Low weight was found in 34.6% of adolescent at the first visit and 5.3% were overweight. Even though, caloric intake was below recommendation, significant increases were observed between first and second trimester for energy, carbohydrates, niacin and zinc (p < 0.001) and for fat, proteins, riboflavin, thiamin, vitamin C, calcium and iron (p < 0.05). From second to third trimester, differences (p < 0.05) were significant only for vitamin A. Between first and third trimester, differences were significant (p < 0.05) for energy, proteins, carbohydrates, niacin, riboflavin, zinc and for thiamin, vitamin A and calcium (p < 0.05). A high proportion of pregnant adolescent did not reach recommendations for energy, folate, calcium and zinc. Food intake pattern did not change significantly among trimesters. Mean total weight gain was 9.2 kg and 0.412 +/- 0.4 g/week. Newborn's mean weight was 3.221 +/- 418 grams. It is concluded that adolescents are at high nutritional risk and deficiency of dietary intake should be followed. Attention should be addressed from the preconceptional period to postpartum in order to improve maternal and fetal conditions. PMID- 14528603 TI - [Profile of nutritional risk in pregnant adolescents]. AB - A sample of 215 adolescents (16.9 +/- 0.7 y; 13-18 y) from a low socioeconomic level of Valencia, Venezuela, at their first trimester of pregnancy were studied in order to identify nutritional risk. Socioeconomic, anthropometrical, hematological and dietary characteristic were assessed. Pregestational weight, actual weight, height, arm circumference (AC), triceps skin fold (TS), fat and muscle area (FA/MA) and pregestational body mass index (PBMI) were determined. Hemoglobin (Hb) and ferritin were measured by colorimetric and ELISA methods. Two 24 hour recalls were obtained. According to age, two groups were created. Pregnant adolescents were grouped by age: Group 1, 13 to 15 years old and Group 2, 16 to 18 years old. Nutritional risk was defined as: Gynecological age lower than 4 years since menarche. PBMI < 19.8 kg/m2, height below 10th Percentile of reference, Hb below 11 g/dL and ferritin < 12 micrograms/L. Almost all the adolescents (96.3%) were in poverty. 83.3% were single, 83.3% were at elementary school before pregnancy but 84.1% were attending house chores at the time of exam. There were significant differences (p < 0.001) for gynecological age and menarche age, but not for anthropometrical, hematological and dietary variables when Group 1 was compared to group 2. 35.3% of the adolescents had a gynecological age lower than 4 years, 23.3% were below 10th percentile for height, 36.3% had a PBMI lower than 19.8 kg/m2, 26.5% were below 10th percentile for arm circumference, anemia was present in 13.7% and low levels of ferritin in 18.4% of the adolescents. Calorie intake was below recommendation in 87.3% and adequacy for vitamin A and C, calcium and zinc were below 2/3 of recommendation in 36.3%. 25.9%, 88.7% and 73.5% of the adolescents respectively. Studied adolescents had a high prevalence of risk factors for nutritional deficiencies (short age, poverty, low weight, anemia and deficient intake). Early assessment of this conditions allows to identify the risk, to establish interventions and to monitor pregnancy evolution. PMID- 14528604 TI - [Nutritional status of folate, vitamin B12 and iron in pregnant adolescents]. AB - Pregnancy in adolescence increases nutritional risk, due to higher demand of nutrients for maternal and fetal growth. This study was aimed to evaluate folate, vitamin B12 and iron status of pregnant adolescents at first trimester of pregnancy. A cross sectional, descriptive study was performed in 122 pregnant adolescents from Valencia, Carabobo state, 1997. Serum and erythrocyte folate and serum vitamin B12 was determined by radioassay; serum ferritin by enzimoimmunoassay; hemoglobin were performed by semi-automated method. Statistical analysis included standard deviation and frequencies. For serum folate 1.7% was found in negative balance and 19.0% at marginal status. For erythrocyte folate, 5.8% was deficient and 1.7% marginal. For serum vitamin B12, 8.3% was deficient and 13.2% marginal. Iron deficiency was found in 19.0% of the adolescents. Prevalence of anemia was of 13.1%, being iron deficiency the main cause. There was high nutritional risk regarding iron status, although iron intakes exceeded the recommendations, but only a small proportion was bioavailable. Prevalence of anemia was lower than reported by other studies and iron deficiency was higher than folic acid and vitamin B12 deficiencies. Pregnant adolescents are at a high biological and nutritional risk. PMID- 14528605 TI - [Plasma amino acids profile of healthy pregnant adolescents in Maracaibo, Venezuela]. AB - One hundred female adolescents (13-18 y) were clinical and anthropometrically studied to select only those with adequate nutrition. Most adolescents belonged to IV socio-economic stratum families (worker class). Height, weight, age, body mass index and medial arm circumference were used as anthropometric parameters. After screening, only 41 non pregnant girls (control) and 42 pregnant girls with adequate nutrition were selected to analyze plasma amino acids. Fasting peripheral venous blood was drawn, and plasma amino acids were analyzed by HPLC. Amino acid concentrations were expressed as umol/L +/- SE. SAS/STAT program was used for statistical analysis. Amino acid values of control adolescent group were found in ranges reported by other investigators, with slight variations, mostly in diminution, presumably due to nutritional, metabolic or genetic conditions of people living in tropical regions. In pregnant healthy adolescents, distributed according to gestational age: < 32 weeks (n = 30) and > 32 weeks (n = 12), a diminution of total molar plasma amino acids was found, by comparing with control values. Ten amino acids (Pro, Gly, Gln, Arg, Ser, Orn, Tau, Leu, Thr and Val) appeared significantively diminished throughout gestation, being Gly. Gln and Arg most affected since earlier weeks. During the 2nd period. Thr and Val increased their grade of affectation; whereas some amino acids values (Orn, Pro and Tau) tended to recuperate. Several of affected amino acids are gluconegoenic, thus, they could be utilized to supply the energy required by the pregnant adolescent against her double stress: the fetus development and her own development. The plasma amino acid values reported in both, healthy non pregnant and pregnant adolescents, could be taken as regional referential profile of plasma amino acids in this poblational group for further research on adolescent and fetal--maternal malnutrition. PMID- 14528606 TI - Comparison of serum concentration and dietary intake of alpha-tocopherol in a sample of urban and rural Costa Rican adolescents. AB - Results from several studies have suggested that vitamin E intake could inhibit the progression of atherosclerotic lesions. Therefore, this study was designed to evaluate Costa Rican adolescents' serum alpha-tocopherol levels and their correlation with Body Mass Index (BMI) and the dietary intake of vitamin E and saturated and polyunsaturated fat. Ninety-five healthy, non-smoking adolescents and non-vitamin supplement users (aged 13-18) from urban and rural areas in San Jose, Costa Rica, were included in this study. Serum levels of lipid adjusted alpha-tocopherol were significantly higher in rural adolescents compared with urban youngsters (4.192 mumol/mmol +/- 0.831 and 3.486 mumol/mmol +/- 0.996, respectively). Likewise, reported mean daily 1000 kcal adjusted-vitamin E intake was higher in rural adolescents than in urban youngsters (9.2 +/- 3.7 mg. 16.0 +/ 8.7 mg). An important correlation was observed between intake of energy from saturated fat and adjusted-alpha-tocopherol serum levels (r = 0.430). Contrariwise, they correlated poorly with dietary alpha-tocopherol (r = 0.273), suggesting that serum is not a good biomarket of intake for alpha-tocopherol. Additionally, our results showed a negative relationship between BMI and adjusted serum vitamin E levels (beta = 0.189; CI 95% -0.153, -0.013). The analysis of vitamin E intake showed that over 25% of adolescents have inadequate intake of the nutrient. This study suggests that nutritionists and pediatricians should encourage alpha-tocopherol intake and monitor the food sources of the nutrient and the adolescents' body weight as part of the strategies aimed at developing a healthy lifestyle. PMID- 14528607 TI - [Comparison of results from two dietary interview methods]. AB - The objective of this study was to compare the food consumption and nutrient intakes obtained through a 24-h dietary recall (DR) and a consumption tendency interview (CTI). Both instruments DR and CTI were applied simultaneously in groups of 264 schoolchildren and 272 adults. Information on age, sex, literacy, occupation, and head of family's social security system was also obtained. Body weight and height were measured and the body mass index BMI was calculated. Food portions and nutrient intakes were described and compared by using means, standard deviation. ANOVA, and product moment Pearson' correlation. No significant differences were detected between both methods with respect to mean consumption of cereals, fruits, meats and sugars. Dairy products and oil consumption were found to be larger according to CTI: 0.6 +/- 1.7 y 0.3 +/- 0.9 portions, respectively (p < 0.01). In terms of nutrient intakes, there were no differences in beta-carotene, vitamin A, vitamin C, iron, and zinc. Mean differences of energy and selected nutrients were lower than 10% of total intake, except by omega-3 fatty acid, cholesterol, vitamin E, and folate. Correlation coefficient between both methods was < 0.4 for most foods and nutrient intakes. Correlation was slightly higher in adults regarding cereals dairy and sugar (approximately 0.5). Both methods DR and CTI produce similar results when applied at the population level, but can be significant different on individuals. The suitability of each method will depend on the objectives of the study. PMID- 14528608 TI - [Effect of dietary calcium during pregnancy and lactation on zinc levels in blood and bone, in rats]. AB - The effect of dietary calcium (Ca) level on maternal zinc (Zn) nutritional status was studied. Female Wistar rats, weighing 250-350 g, were fed during pregnancy and lactation with an experimental diet containing/100 g different levels of calcium: 0.2 g (low calcium: LCa), 0.6 g (normal calcium: NCa) or 0.9 (high calcium: HCa). Maternal blood samples were drawn from the tail at delivery and at the end of lactation. Laboratory determinations were: Zn in whole blood (WB) at delivery and weaning; Zn (ZnF) and Ca (CaF) in the ashed femur at weaning. The results (mean +/- SEM) were: ZnWB (microgram/ml) at delivery and weaning: LCa: 8.73 +/- 1.05; 12.8 +/- 2.02; NCa: 3.49 +/- 0.19; 3.73 +/- 0.37; HCa: 3.21 +/- 0.19; 3.85 +/- 0.27. CaF (mg/100 mg): LCa: 19.2 +/- 0.8; NCa: 21.4 +/- 0.6; HCa: 20.4 +/- 1.1. ZnF (microgram/100 mg): LCa: 30.2 +/- 0.9; NCa: 24.1 +/- 0.3; HCa: 24.1 +/- 0.9. ZnWB was significantly higher in LCa (p < 0.0001) regarding NCa and Hca. ZnF showed an increase and CaF a decrease in LCa regarding NCa and HCa (p < 0.0001). There were no significant differences in ZnWB, ZnF and CaF between NCa and HCa These results show that: there was no detrimental effect when dietary Ca content was increased by 50% above the normal requirements of the rat.; low dietary Ca during pregnancy and lactation produced an increase of Zn utilization, reflected in maternal blood Zn and in ZnF content. PMID- 14528609 TI - [Occurrence of Listeria monocytogenes in pre-sliced vacuum-packaged]. AB - There is scarce information in Brazil and other South American countries about the occurrence of Listeria monocytogenes in food, mainly refrigerated ready-to eat products. The consumption of sliced vacuum-packaged meat products has increased in the last few years. Nevertheless, a complete assessment of the risk associated with L. monocytogenes in these products is still necessary. Because of the production and storage characteristics of these products, they can be considered potential vehicles for L. monocytogenes to humans, mainly immunocompromised, elderly, and pregnant women. The objectives of this study was to evaluate the population of L. monocytogenes in salami, a ready-to-eat meat product with extended shelf life, acquired in retail stores in Sao Paulo-Brazil. The three-tube most probable number technique was used and the methodology was that from Health Canada. Strains were biochemically identified and serotyped. Among the 45 samples, 3 (6.7%) harboured 9.2 MPN/g of L. monocytogenes and the others < 0.3 MPN/g. All the strains belonged to serotypes 1/2a and 1/2b, the most frequent serotypes found in food everywhere. Even being low, the population of L. monocytogenes found in this product could be a cause of concern to public health authorities as it can pose a threat to population at risk. This contamination highlights the importance of implementing systems like HACCP to assure safe products to consumers. PMID- 14528610 TI - [Physicochemical changes in starch during corn alkaline-cooking in varieties with different kernel hardness]. AB - Changes in starch characteristics caused by alkaline-cooking in two corn genotypes (hard and soft) were studied in the present work. Color, pH, pasting properties (Brabender viscoamylograph), water sorption, solubility, swelling power and thermal analysis (Differential scanning calorimetry) were determined in starches previously extracted, from raw and alkaline-cooked hard and soft corns. A reduction in maximum viscosity peak, temperature at the endothermic peak and swelling power at 90 degrees C, and an increase in solubility was observed in starches obtained from both hard and soft corn after alkaline-cooking. Starch from hard corn showed an increment in initial pasting temperature after alkaline cooking. Alkaline-cooking induced higher modifications in hard than in soft corn starch, probably as a result of its endosperm type and grain hardness, although optimum cooking times were used for each corn genotype. PMID- 14528611 TI - [Production and partial characterization of beta-galactosidase from Kluyveromyces lactis grown in deproteinized whey]. AB - The purpose of this work was to optimize the beta-galactosidase production by Kluyveromyces lactis, applying the Surface Response Methodology (SRM) and using deproteinized whey as fermentation medium. An Orthogonal Central Compound Design (OCCD) was used without repetition, with four factors: temperature, pH, agitation speed and fermentation time. Then, enzyme activity (U/ml) as response variable was used. Thirty trials in twenty-five treatments, with six repetitions at the central point, were carried out, in a New Brunswick Bioflo 2000 fermentor with a volume of 2 liters. The deproteinized whey obtained by thermocoagulation was chemically analyzed. The results were: moisture 93.83%, total solids 6.17%, protein 0.44%, lactose 4.85%, acidity 0.43% and pH 4.58. The best conditions in the enzyme production were: temperature 30.3 degrees C, pH 4.68, agitation speed 191 r.p.m. and fermentation time 18.5 h. with an enzyme production of 8.3 U/ml. The degree of purification obtained was 7.4 times and the yield was 50.8%. The purified enzyme had an optimum temperature of 60 degrees C and a pH of 6.2. This work shows that the yeast Kluyveromyces lactis grown in deproteinized whey is able to produce the enzyme beta-galactosidase and SRM can be used in the fermentology processes, specifically in determining the best suitable operation conditions. PMID- 14528613 TI - [Development of an optimized formulation of damask marmalade with low energy level using Taguchi methodology]. AB - The goal of this present study was the development of an optimized formula of damask marmalade low in calories applying Taguchi methodology to improve the quality of this product. The selection of this methodology lies on the fact that in real life conditions the result of an experiment frequently depends on the influence of several variables, therefore, one expedite way to solve this problem is utilizing factorial desings. The influence of acid, thickener, sweetener and aroma additives, as well as time of cooking, and possible interactions among some of them, were studied trying to get the best combination of these factors to optimize the sensorial quality of an experimental formulation of dietetic damask marmalade. An orthogonal array L8 (2(7)) was applied in this experience, as well as level average analysis was carried out according Taguchi methodology to determine the suitable working levels of the design factors previously choiced, to achieve a desirable product quality. A sensory trained panel was utilized to analyze the marmalade samples using a composite scoring test with a descriptive acuantitative scale ranging from 1 = Bad, 5 = Good. It was demonstrated that the design factors sugar/aspartame, pectin and damask aroma had a significant effect (p < 0.05) on the sensory quality of the marmalade with 82% of contribution on the response. The optimal combination result to be: citric acid 0.2%; pectin 1%; 30 g sugar/16 mg aspartame/100 g, damask aroma 0.5 ml/100 g, time of cooking 5 minutes. Regarding chemical composition, the most important results turned out to be the decrease in carbohydrate content compaired with traditional marmalade with a reduction of 56% in coloric value and also the amount of dietary fiber greater than similar commercial products. Assays of storage stability were carried out on marmalade samples submitted to different temperatures held in plastic bags of different density. Non percetible sensorial, microbiological and chemical changes were detected after 90 days of storage under controlled conditions. PMID- 14528612 TI - [Elaboration of "cotija" type cheese made of whole milk and chickpea (Cicer arietinum L.) mixture]. AB - The objective [corrected] of this work was to elaborate "cotija" type cheese prepared with whole milk and chickpea (Cicer arietinum L.), inoculated with S. Thermophilus and to compare its physicochemical, microbiological and sensorial characteristics with a commercial cheese. Two mixture were selected 70:30 and 80:20 (whole milk:chickpea paste). Both mixture were ground, pasteurized and inoculated with. S. Thermophilus, to develop taste and aroma. Two rennet concentrations (1:10,000 and 2:10,000), lactic acid and 50% CaCl2 solution were used in both products. Results showed that cheese made with 70:30 mixture and 1:10,000 rennet concentration gave a weak curde and 12% of total solids were lost; in the same way as 70:30 mixture and 2:10,000 rennet concentration. Nevertheless, cheese made with 80:20 mixture and 10,000 or 2:10,000 rennet concentration showed hard consistency of its curd. Addition of 10% sodium chloride solution to he 80:20 mixture cheese, made better whey drain, and a "cotija" type cheese with similar sensory characteristics as a whole milk cheese, was obtained. Chemical and physical analysis of the extended "cotija" type cheese showed 14.3 +/- 0.42 and 15.5 +/- 0.21 g/100 g of protein and lipid content respectively. Microbiologically, the extended "cotija" type cheese is pathogenic microorganisms free for human consumption. Sensory evaluation of the chickpea "extended" cheese showed a 80% acceptance while the acceptance of the whole milk commercial cheese was 90%. PMID- 14528614 TI - Two-group experimental designs. PMID- 14528615 TI - Evaluation of individually tailored interventions on exercise adherence. AB - This study's purpose was to test the effects of a reversal theory-driven individualized exercise prescription on exercise consistency, weight, percentage body fat, and exercise motivation for a group of faculty, students, and staff at a southwestern university. Participants were randomly assigned to 5-week education plus monitoring or monitoring-only treatment groups. The hypothesis was that participants in the education plus monitoring group would have more consistent exercisers, higher exercise motivation scores, and greater overall weight loss and decrease in percentage of body fat than would those in the monitoring-only group. Consistent exercisers will score higher than inconsistent exercisers on exercise motivation scores. More participants were exercising at recommended levels in the education plus monitoring group than in the monitoring only group after the study intervention. Consistent exercisers had significantly higher motivation scores than did inconsistent exercisers. Thus, individualized exercise prescriptions using reversal theory can be beneficial in promoting a consistent exercise program. PMID- 14528616 TI - Communicating end-of-life preferences. AB - The purpose of this survey was to explore how adults communicate their end-of life preferences. Face-to-face interviews were conducted with 119 community dwelling adults who had previously engaged in conversations about their end-of life preferences. Factors that made it easier to initiate the discussion included having personal experience with illness or death (24.4%), being straightforward (24.4%), or having someone else facilitate the discussion (11.8%). Most described vague end-of-life preferences such as not wanting any machines (41.2%) or heroics (34.5%). Although 22.7% reported using a living will to make their preferences clear, only 5.9% mentioned repeating or reinforcing their preferences. In all, 21% had discussed their end-of-life preferences with their physicians. These findings show discussions about end-of-life preferences frequently lack the clarity and detail needed by significant others and health care providers to honor the preferences. Routine dialogue with health care providers and significant others about end-of-life preferences might provide greater clarity and comfort. PMID- 14528617 TI - The substance of a support group. AB - This ethnographic research constitutes a study of the culture of a family-run advocacy organization for families of children and youth with mental health needs. Data collection includes 703 pages of interview transcripts, observations, field notes, and archival material collected by the principle investigator during 2.5 years of participant observation with this support group. This article provides the context for the study and describes the support group's cultural ideologies and cultural forms that are a concrete expression of those ideologies: There are four major categories of cultural forms as follows: symbols, language, narratives or stories, and practices. Manifestations of each form are discussed and described. The proliferation of support and advocacy groups in the mental health arena promises a rich source of understanding of the needs of families in distress as well as inspiration for new and practical interventions. PMID- 14528618 TI - Clinical reasoning in experienced nurses. AB - As an essential component of nursing practice, clinical reasoning is used to assimilate information, analyze data, and make decisions regarding patient care. Little is known about the reasoning strategies of experienced nurses who are not yet experts. This qualitative descriptive study explored the cognitive strategies used by experienced nurses as they considered assessment findings of assigned patients. To date, few studies of nurses' clinical reasoning have been conducted in a practice setting during actual patient care. A small group research design was employed using the think-aloud (TA) method with protocol analysis. A total of 15 experienced nurses were asked to "think aloud" about patient assessment findings. Data were audiotaped, transcribed, and analyzed using the three steps of protocol analysis. The results suggest that experienced nurses used a conceptual language to reason about assessment findings and used heuristics to reason more quickly and efficiently. PMID- 14528619 TI - Reworking professional nursing identity. AB - In spite of professional socialization through nursing education programs, new graduates experience stress as they become working professionals. This grounded theory study explores experienced nurses' perceptions of how they became professional. The central problem for nurses was dissonance between expectations and experiences; they addressed this through an iterative, three-stage process of reworking professional identity. The stages of this process are assuming adequacy, realizing practice, and developing a reputation. Iterations of this process occur as new discrepancies are noticed, enhanced awareness dawns, practice changes, learning is undertaken, or experienced nurses become relative novices in another work area. Nurses move through stages more quickly and at different levels with each iteration. Three contextual factors influence the process: expectation; perception of the status accorded by others to nursing; and supportiveness by acceptance, assistance, and advocacy from others in the workplace. These findings expand knowledge about professional socialization and how nurses themselves understand developing professional identity. PMID- 14528620 TI - Explanations for improvement in both experimental and control groups. AB - A true experimental design with random assignment to groups protects against false causal inferences that could be made when both the treatment and control groups change because of factors such as testing effects, reactivity, contamination, maturation, history, and other measurement effects. The occurrence of these phenomena, however, provides interesting information about factors affecting health care attitudes, knowledge, and behavior change, which can interfere with a nursing study's ability to demonstrate an experimental effect. In this article, we discuss these design threats, illustrate them with examples from recent health research, and suggest strategies for decreasing them in clinical nursing studies. PMID- 14528621 TI - The evolutionary roots of our environmental problems: toward a Darwinian ecology. AB - It is widely acknowledged that we need to stabilize population growth and reduce our environmental impact; however, there is little consensus about how we might achieve these changes. Here I show how evolutionary analyses of human behavior provide important, though generally ignored, insights into our environmental problems. First, I review increasing evidence that Homo sapiens has a long history of causing ecological problems. This means that, contrary to popular belief, our species' capacity for ecological destruction is not simply due to "Western" culture. Second, I provide an overview of how evolutionary research can help to understand why humans are ecologically destructive, including the reasons why people often overpopulate, overconsume, exhaust common-pool resources, discount the future, and respond maladaptively to modern environmental hazards. Evolutionary approaches not only explain our darker sides, they also provide insights into why people cherish plants and animals and often support environmental and conservation efforts (e.g., Wilson's "biophilia hypothesis"). Third, I show how evolutionary analyses of human behavior offer practical implications for environmental policy, education, and activism. I suggest that education is necessary but insufficient because people also need incentives. Individual incentives are likely to be the most effective, but these include much more than narrow economic interests (e.g., they include one's reputation in society). Moralizing and other forms of social pressure used by environmentalists to bring about change appear to be effective, but this idea needs more research. Finally, I suggest that integrating evolutionary perspectives into the environmental sciences will help to break down the artificial barriers that continue to divide the biological and social sciences, which unfortunately obstruct our ability to understand ourselves and effectively address our environmental problems. PMID- 14528623 TI - International Nurses Day 2003. PMID- 14528622 TI - Snake bioacoustics: toward a richer understanding of the behavioral ecology of snakes. AB - Snakes are frequently described in both popular and technical literature as either deaf or able to perceive only groundborne vibrations. Physiological studies have shown that snakes are actually most sensitive to airborne vibrations. Snakes are able to detect both airborne and groundborne vibrations using their body surface (termed somatic hearing) as well as from their inner ears. The central auditory pathways for these two modes of "hearing" remain unknown. Recent experimental evidence has shown that snakes can respond behaviorally to both airborne and groundborne vibrations. The ability of snakes to contextualize the sounds and respond with consistent predatory or defensive behaviors suggests that auditory stimuli may play a larger role in the behavioral ecology of snakes than was previously realized. Snakes produce sounds in a variety of ways, and there appear to be multiple acoustic Batesian mimicry complexes among snakes. Analyses of the proclivity for sound production and the acoustics of the sounds produced within a habitat or phylogeny specific context may provide insights into the behavioral ecology of snakes. The relatively low information content in the sounds produced by snakes suggests that these sounds are not suitable for intraspecific communication. Nevertheless, given the diversity of habitats in which snakes are found, and their dual auditory pathways, some form of intraspecific acoustic communication may exist in some species. PMID- 14528624 TI - Developing leadership abilities in nursing. PMID- 14528625 TI - Independent geriatric nurse practitioner. PMID- 14528626 TI - Counselling in family welfare. PMID- 14528627 TI - Dr Kelly's death and the subsequent political tailspin has blotted out almost every other conceivable element of public debate. What is happening to the summer discussion about health? PMID- 14528628 TI - Data protection problems. PMID- 14528629 TI - Exchange rates. PMID- 14528631 TI - On the front line. PMID- 14528630 TI - Regional accents. The RCN policy unit compares NHS plans across the UK. AB - All three NHS plans that form the modernization agenda can be characterised by three themes: Centralization Partnership Culture change. Despite the absence of a Northern Ireland plan, the issues and themes that emerge in the other three plans are prevalent in Northern Ireland too. CENTRALISATION: The tendency to centralize has led to a greater involvement of health ministers, rather than civil servants, in the day-to-day running of the health services, but ministers are preparing to hand over operational control of new policies. PARTNERSHIP: Partnership underpins many of the new structural arrangements, with an emphasis on increased inter professional working and education, but merged health and social care structures create funding tensions. CULTURE CHANGE: Culture change focuses on creating patient-centred care and dismantling the power of the health professions, while the curbing of professional autonomy is central to enhancing and improving patients' overall experience of the NHS. The cumulative effect of these developments is likely to lead to increasing debate about the future funding, provision and accountability of the NHS as regional and country differences continue to develop. PMID- 14528632 TI - Pick of the crop. PMID- 14528633 TI - Exploring Alexis' model: Part two. Valuing resources. PMID- 14528634 TI - Managing mistakes and challenges. PMID- 14528636 TI - Taking it to the top. PMID- 14528637 TI - True professionals. Interview by Frances Pickersgill. PMID- 14528638 TI - The price is right. PMID- 14528639 TI - Speaking for ourselves. PMID- 14528640 TI - Opening the door to healthcare assistants and support workers. PMID- 14528641 TI - Nursing older adults with mental health problems: therapeutic interventions--Part 1. PMID- 14528642 TI - Myocardial infarction and older people. AB - Nurses have a central role to play in the care of older people who have suffered a myocardial infarction. This article points out that such patients may not present with typical symptoms so it is important to be aware of signs that indicate problems. It suggests that following diagnosis it is vital that older people are offered evidence-based treatment and that secondary prevention and rehabilitation are considered. PMID- 14528643 TI - Assessing pulse in older people. PMID- 14528644 TI - A better deal for neonatal care at last. AB - All those concerned with the care of premature and sick newborn babies have been calling for a review of neonatal services for almost a decade. Now, at last, the long-awaited Department of Health (DoH) review of neonatal intensive care services in England has been published. PMID- 14528645 TI - New guidelines on infant feeding in the first 12 months of life. AB - Paediatric dietitian Judy More discusses several recent directives and guidelines on infant feeding in the first year of life. These include recommendations that babies should be fed exclusively on breast milk for their first six months and that soy formulae should normally be avoided for babies under six months old. There is also specific new advice on restricting salt intake for infants. PMID- 14528646 TI - Paediatric group position statement on breast feeding and weaning on to solid foods. PMID- 14528647 TI - Paediatric group position statement on the use of soya protein for infants. AB - Breast feeding should be strongly encouraged as providing the safest, most nutritionally adequate form of feeding for most infants. Dietitians should discourage the use of soya protein in children with atopy or cow's milk allergy in the first six months of life to avoid sensitisation to soya protein and exposure to phytoestrogens while organ systems remain at their most vulnerable. This would include soy infant formula and soya products such as desserts etc. When a soy-based infant formula is used, parents should be informed of current findings relating to phytoestrogens and health and on the clinical need for soy formula. Any parent choosing to refuse soya for their infant should be supported in their decision. More research into the long-term effects of phytoestrogen exposure in infants is needed and into whether any adverse effects are dose related. This position statement will be updated as further evidence becomes available. PMID- 14528648 TI - Health and fitness series--4. Getting children to develop a healthy relationship with food. AB - The author, a dietitian, discusses the trend in the UK towards children developing unhealthy relationships with food. This is manifested as overeating leading to overweight or obesity, or a desire, even among younger children, to follow slimming diets that can adversely affect growth and development. Rather than food restriction, a healthy relationship with food involves young people being able to self-regulate their intake and enjoy all the components of a balanced diet. The author emphasises the importance of mothers as role models in helping their children develop a healthy relationship with food. Physical activity should be encouraged, preferably for the whole family. PMID- 14528649 TI - Managing gastro-oesophageal reflux in infants and children. AB - Dr Beattie describes the different types of gastrooesophageal reflux and the investigations that may be needed. Management in infants and children is discussed, from simple measures through to more complex medications and, in a minority of cases, surgery. Mild reflux occurs in 50% of all babies and most cases are managed by health visitors and general practitioners. Simple strategies, including feeding advice and reassurance about the natural history, are usually sufficient. Feed thickeners or an anti-reflux milk help in selected cases. There is a need, however, for careful clinical assessment of cases and consideration of the differential diagnosis. It is important that resistant or difficult cases are assessed by a paediatrician with an interest in reflux. PMID- 14528650 TI - Screening at school entry: vision, hearing and growth. AB - Tam Fry of the Child Growth Foundation criticises the new recommendations for screening at school entry in the 2003 edition of Health for all Children. He argues that proposals to reduce preschool surveillance make the school entry screening even more important for detecting deviations from the normal, especially in vision, hearing and growth. This surveillance needs to be rigorously performed, yet a shortage of school nurses and optometrists makes this less likely to happen. The author argues that even though there may be insufficient evidence for retaining current preschool screening programmes, there is also insufficient evidence to show that severely curtailing them will not cause harm. PMID- 14528652 TI - Hospitals nationwide brace for Oct. 16 HIPAA transactions conversion. PMID- 14528651 TI - Constipation in children: avoiding hospital admissions by the use of a specialist community nurse. AB - OBJECTIVE: To review paediatric admissions with a primary diagnosis of constipation to see whether some could have been managed in the community instead. METHOD: A review of the medical notes of all patients admitted with a primary diagnosis of constipation to the children's ward of a district general hospital over a 12-month period. RESULTS: Of 41 admissions (19 girls and 22 boys, age range 6 weeks to 12 years), the average length of stay was less than two nights. CONCLUSION: The short duration of hospital stay implies rapid improvement. It is likely that many of these children could have been managed in the community if suitable resources, such as a community nurse specialising in constipation, were available. PMID- 14528653 TI - Hospital CEO indicted under the anti-kickback statute for physician recruiting activities. PMID- 14528654 TI - Get into the game. PMID- 14528655 TI - [Smoking and preoperative fasting--are there evidence-based guidelines?]. AB - Over the last years several clinical studies have modified the guidelines for preoperative fasting to reduce the risk of pulmonary aspiration. In most western countries the following guidelines are accepted: for clear liquids 2 hours, breast feeding 4 hours, small meals and breast milk substitutes 6 hours, heavy meals 8 hours. Since preoperative smoking is acknowledged as a risk factor, it should be ceased in most clinics 6 hours before induction of anaesthesia, as well. Smoking, however, does not increase the risk of pulmonary aspiration, as is often maintained, but increases the risk of postoperative pulmonary complications. To reduce the risk of perioperative pulmonary complications, cessation of smoking is necessary 8 weeks before operation. Stopping smoking only a few days before operation and anaesthesia even tends to increase the risk of pulmonary complications. Regarding cardiac complications, cessation of smoking 12 hours before anaesthesia is sufficient to reduce the incidence of cardiac ischaemia. PMID- 14528656 TI - [Mesenteric traction syndrome during the operation of aneurysms of the abdominal aorta--histamine release and prophylaxis with antihistaminics]. AB - Mesenteric traction syndrome occurs during abdominal surgery and is described as sudden tachycardia, hypotension and flush. Among other etiological factors, eventeration or mesenteric traction of the small intestine may cause histamine release from mesenteric mast cells. Therefore, our hypothesis was that mesenteric traction syndrome could be positively influenced by prophylactic administration of H1- and and H2-antihistamines. Seventeen male patients (ASA groups III-V, 48 78 years old) were investigated in a randomised double blind study during elective abdominal aortic aneurysm (AAA) repair; which, in our opinion, is one of the most standardised surgical procedures. Eight patients had pre-anaesthetic prophylaxis with 0.1 mg/kg BW dimetindene (H1-receptor antagonist) plus 5 mg/kg BW cimetidine (H2-receptor antagonist) diluted with 100 ml 0.9% NaCl, while 9 patients received a placebo (100 ml 0.9% NaCl). Anaesthesia and invasive haemodynamic monitoring were standardised in all patients. Haemodynamic parameters, plasma histamine concentrations and clinical symptoms were determined one min after skin incision (HS), and 5 and 20 min after mesenteric traction (5' EV and 20' EV). Statistical analyses were performed using the Student's t-test, the Mann-Whitney-U-test for continuous data and Chi2-test for incidences. The incidence of histamine release was 55.5% (5/9) in the placebo group vs. 37.5% (3/8) in the antihistamine group (p > 0.05, Chi2-test). Plasma histamine levels (mean +/- SD) were higher in the placebo group than in the antihistamine group at 5 and 20 min after mesenteric traction, but there was no statistical significance. Arrhythmias were significantly more frequent in the placebo group (6 times) than in the antihistamine group (none) (p = 0.005 Chi2-test). Systolic blood pressure was not statistically different between the groups (e.g. 5 min after mesenteric traction, mean +/- SD; placebo 111 +/- 20 mm Hg vs. antihistamines 119 +/- 35 mm Hg). In the placebo group, however, the haemodynamics only stabilised 5 min after mesenteric traction when anaesthetic gas concentration was repeatedly reduced and vasopressor/volume administration was increased (placebo group = 20 times vs. antihistamine group = 8 times (p = 0.001, Chi2-test). From these results we conclude that prophylactic administration of antihistamines reduces in particular the incidence of arrhythmias and the number of stabilising measures during mesenteric traction. Prophylaxis with H1- and H2-antihistamines may therefore be of perioperative benefit and should be considered in AAA surgery. PMID- 14528657 TI - [Continuous biochemical tissue monitoring during acute hypoxia]. AB - Oxygen deficiency during critical illness is known to cause profound changes in cellular metabolism with subsequent organ dysfunction. Clinical treatment in these patients is focussed on rapid reoxygenation to avoid a prolonged impaired synthesis of cellular high-energy phosphates (ATP). The effect of this therapeutical intervention on the level of the cell, however, has not yet been objectivized. The aim of the present experimental study was to biochemically monitor different tissues during hypoxia and reoxygenation using in vivo microdialysis. Eighteen adult male CD-rats (412-469 g; Ivanovas, Kisslegg, Germany) were normoventilated under general anaesthesia (FiO2 = 0.21). Ten were then subjected to a period of hypoxia (FiO2 = 0.1, 40 min) and reoxygenated with FiO2 = 0.21, while eight control animals were continuously ventilated with FiO2 = 0.21. In addition to invasive haemodynamic monitoring, biochemical tissue monitoring was performed using CMA 20 microdialysis probes, which were inserted into the muscle (m), subcutaneous space (s), liver (l) and peritoneal cave (p) with semicontinuous analyses of lactate and pyruvate at intervals of 15 minutes. Hypoxia induced a significant decrease in mean arterial pressure compared to the control group (p < 0.05). At the same time significant increases in blood lactate (12.3 + 4.1 mmol/l (hypoxia) vs. 1.5 +/- 0.3 mmol/l (control); p < 0.05) and in negative base excess (17.3 + 7 mmol/l (hypoxia) vs. 2.6 + 1.8 mmol/l (control), p < 0.05) occurred. Compared to unchanged levels in the control animals, the interstitital lacate/pyruvate ratio in the investigation group rose to significantly higher values (455 + 199% of baseline (m), 468 + 148% (p), 770 + 218% (l) and 855 + 432% (s) (p < 0.05). An immediate return to the baseline values after the start of reoxygenation was noted in the L/P ratio during the observation period. Using microdialysis, it was possible to objectify the effect of oxygen deficiency and restoration on tissue metabolism. Regarding clinical and preclinical practice, microdialysis monitoring should be performed to include biochemical cellular effects as an additional target for therapeutical interventions. PMID- 14528658 TI - Beyond point and shoot: children's developing understanding of photographs as spatial and expressive representations. PMID- 14528659 TI - Probing the adaptive significance of children's behavior and relationships in the school context: a child by environment perspective. PMID- 14528660 TI - The role of letter names in the acquisition of literacy. PMID- 14528661 TI - Early understandings of emotion, morality, and self: developing a working model. PMID- 14528662 TI - Working memory in infancy. PMID- 14528663 TI - The development of a differentiated sense of the past and the future. PMID- 14528664 TI - The development of cognitive flexibility and language abilities. PMID- 14528665 TI - A bio-social-cognitive approach to understanding and promoting the outcomes of children with medical and physical disorders. PMID- 14528666 TI - Expanding our view of context: the bio-ecological environment and development. PMID- 14528667 TI - Pathways to early literacy: the complex interplay of child, family, and sociocultural factors. PMID- 14528668 TI - Some lobbying tactics have pols seeing red. PMID- 14528669 TI - Consumer-directed health care: too good to be true? PMID- 14528670 TI - Pharmacy benefit starts to reflect push for consumer-directed care. PMID- 14528671 TI - Should benefit to individuals outweigh good for groups? PMID- 14528672 TI - This isn't the first attempt to shift cost to employees. PMID- 14528673 TI - Balancing the health care scorecard. PMID- 14528674 TI - Too early to write obituary for patients' bill of rights. PMID- 14528675 TI - MC strategy: create competition in expanding biotech field. PMID- 14528676 TI - [Investigation on occurrence of lycium barbarum pests and its natural enemies at Ningxia]. AB - OBJECTIVE: To provide the scientific basis for controlling Lycium barbarum pests. METHODS: The kinds and number of the pests and its natural enemies were investigated at fixed plants. RESULT: There were 38 species of Lycium barbarum pests and 23 species of natural enemies, the occurrence of them were closely related. CONCLUSION: Natural enemies can control Aphis and Paratrioza effectively. PMID- 14528677 TI - [The effects of culture ways on the callus growth and saponin content of Panax quinquefolius]. AB - The study was focused on the effects of solid culture and suspension culture on the callus growth and saponin content of callus from the buds of Panax quinquefolius L. The results showed that the growth carves were all "S" shape under two cultural ways. Compared with the solid culture, the whole course of suspension cultrure was advanced five days, and the dry weight of callus was higher than that under solid culture. The content of the saponin, amine acid and starch also. The studies showed that it was possible to produce saponin qinsenosides by large-scale of Panax quinquefolius L. PMID- 14528678 TI - [Comparative study on content of chlorogenic acid in Lonicera japonica and L. macranthoides]. AB - OBJECTIVE: To exploit and utilize reasonably the abundant natural resources by appraising the medicinal purposes of Lonicera macranthoides Hands-Mazz. through the content of the useful chemical constitutents chlorogenic acid. METHOD: The content of chlorogenic acid in the flowers of L. macranthoides Hands-Mazz in Human and L. japonica Thunb. in Henan and Shandong was compared by HPLC. RESULT: The contents of chlorogenic acid were 4.00% and 4.52% in L. macrathoides from Longhui and Xinning County of Hunan respectively. The contents were 2.20% and 2.46% in L. japonica from Shangdong and Henan respectively. CONCLUSION: The content of chlorogenic acid in L. macranthoides was higher than that in Lonicera japonica. PMID- 14528679 TI - [Microscopical identification of roots and rhizome of six species of plants from viola in the region of Nanjing]. AB - Microscopical identification of 6 species of plants from Viola in Nanjing was made, and the distinguished microscopical characteristics of them were found. The key was also listed on, which can provide scientific data for identification and exploitation of Viola plants. PMID- 14528680 TI - [Inhibiting effect of ethanol extract from Achyranthes bidentata on A beta 42 aggregation]. AB - OBJECTIVE: To observe inhibiting effect of ethanol extract from Achyranthes bidentata (Nx-E) on A beta 42 on (beta-amyloid protein 42) aggregation and fibril formation. METHODS: A beta 42 and Nx-E were incubated in 37 degrees C, 5%, CO2 incubator first, fibril formation was monitored by thioflavine-T fluorescence and was observed with transmission electron microscopy. RESULTS: At 7 days, 1 microgram.microL-1 A beta 42 fluorescence intensity was significantly increased, and much amyloid-like fibril was observed. A beta 42 co-incubated with 15 and 30 micrograms.microL-1 Nx-E for 7 days, their fluorescence intensities were remarkably reduced, few fibrils but an amorphous material were often observed. CONCLUSIONS: After 1 microgram.microL-1 A beta 42 was incubated alone up to 7 days, A beta 42 aggregated and amyloid-like fibril formed. 15 and 30 micrograms.microL-1 Nx-E can prevent A beta 42 from polymerizing to some extent. PMID- 14528682 TI - [Pharmacodynamics study of Fuyuan oral liqiuid on enhancing intestinal function]. AB - OBJECTIVE: To observe the effect of Fuyuan oral liquid (FYOL) on mice and rats intestinal function. METHOD: The effect of FYOL on the carbo-dust drive distance in mice whose intestinal function is normal or weakened by atropine was determined. The effect of FYOL on rats intestional movement curves and mesenteric microcirculation was determined. The effect of FYOL on the inflammatory exudation due to aceti acid in mice was determined. RESULT: FYOL increased mice intestinal drive distance regardless of function normal or weakened by atropine. FYOL increased rats intestional peristalsis intensity but not fequency and ameliorates mesenteric microcirculation. FYOL inhibited mice inflammatory exudation by acetic acid. CONCLUSION: FYOL can accelerate intestinal peristalsis and ameliorate mesenteric microcirculation and anti-inflammation. PMID- 14528681 TI - [Protective effects of tanshinone IIA on injured primary cultured rat hepatocytes induced by CCl4]. AB - OBJECTIVE: To investigate the effect of tanshinone IIA on carbon tetrachloridlde (CCl4) intoxicated primary cultured rat hepatocytes. METHODS: The rat hepatocytes were separated by perfusion with collagenase solution. Tanshinone IIA was added to the cultured medium after incubating the rat hepatocytes for 36 hours. At the same time, CCl4 was added to induce hepatocytes injury for 1 h. The levels of ALT, LDH, NO, SOD, MDA and cell viability after cultivation for 3 h were determined respectively. RESULT: Tanshinone IIA significantly improved cell viability, inhibited the decrease of SOD activity and the increase of ALT, LDH, MDA, NO levels CONCLUSION: Tanshinone IIA protected effectively the primary cultured rat hepatocytes against CCl4-induced injury. PMID- 14528683 TI - [Pharmacodynamics study on "tao hua zhi xie granule"]. AB - The anthraco-propulsion and the drug induced mice diarrhea models were used to observe the intestine propulsion and the anti-diarrhea effect of "Tao Hua Zhi Xie Granule" (THZXG), and the anti-inflammation effect of THZXG was studied. The results showed that THZXG could obviously reduce the incidence and frequency of the mice diarrhea induced by Folium Sennae and castor-oil, and propelling movement of mice small intestine after hypodermic injection of neostigmine. The actions were acted in a dose-dependent manner. 11.70 and 17.55 mg/kg THZXG(ig) could also inhibit the increased permeability of intraperitoneal capillary induced by acetic acid in mice. PMID- 14528684 TI - [Studies on antiendotoxin action of total saponins from radix Bupleuri]. AB - OBJECTIVE: To study antiendotoxin activities of total saponins from Radix Bupleuri. METHOD: The antiendotoxin activities were demonstrated by Limulus Amebocyte Lysate test (LAL test) in vitro, the fever model induced by endotoxin in rabbits and the model induced by LPS in BCG-primed mice. RESULTS: Total saponins from Radix Bupleuri still had antiendotoxin effect even when it (50 mg.ml-1) was diluted to 32 times by LAL test in vitro. The fever experiment results showed the average rising temperature of BCS + LPS, group, model group and BCS group was 0.75 degrees C, -0.25 degrees C and -0.37 degrees C respectively. In the experiment of the poison induced by LPS, fatal rate of model group was 90.0%, that of BCS + LPS group was 13.3%, there was a significant difference (P < 0.01). CONCLUSION: The result showed that the total saponins in Bupleuri had the strongest antiendotoxin action. PMID- 14528686 TI - [Herbalogical study on tianxiong]. AB - According to textual study of herbal drugs and modern scientific research on Tianxiong, the long biennial root tuber losing reproductive capacity of Aconitum carmichaeli Debx. is regarded as the orthodox medicinal herbs through the ages. PMID- 14528685 TI - [Studies on supercritical CO2 fluid extraction for ginkgolic acids in the epicarp of Ginkgo biloba]. AB - The technology of supercritical CO2 fluid extraction for ginkgolic acids in the epicarp of Ginkgo biloba L. was studied. The effect of pressure, temperature and extraction time on the yield of the ginkgolic acids was explored. The optimum conditions for supercritical CO2 fluid extraction was 30 MPa, 45 degrees C, extraction time 6 h and the flow rate of CO2 2L/min. The ginkgolic acids were determined by HPLC. Supercritical CO2 fluid extraction exceled the traditional solvent extraction in high yield, high purification and easy operation. PMID- 14528687 TI - [Review in the studies on tannins activity of cancer prevention and anticancer]. AB - This paper reviewed the biological activities of tannins in cancer prevention and anticancer, and mainly discussed related mechanisms. The results suggest that tannins, whether total tannins or pure tannin compound, have remarkable activity in cancer prevention and anticancer. It has wealthy foreground for developing new cancer prevention agents and/or new anticancer drugs screening among tannin compounds. PMID- 14528688 TI - [Observation of blooming habits of Panax notoginseng]. AB - This paper reported the whole process of blooming habits of Panax notoginseng and pollens scattering regularities in Wenshan, Yunnan province. It can provide reference for the pollenation of hybridization. PMID- 14528689 TI - [Resources of medicinal pteridophyta in Fengyang Mountain and Baishanzu Nature Reservation]. AB - Natural environment of Fengyang Mountain and Baishanzu Nature Reservation is superiority. The vegetation is preserved well and the plant resources is rich. There are 79 species medicinal pteridophyta in this area. According to potency, it can be divided into 16 types, such as medicines for relieving exterior syndrome, medicines for clearing away heat, antirheumatic, medicines for inducing diuresis, medicines for expelling the parasites, hemostasis medicines, medicines for activating blood circulation to remove blood stasis, medicines for restoring vital energy, medicines for calming the liver to stop the wind, etc. PMID- 14528690 TI - [The computer image processing and cluster analysis on Fructus Cnidii from different habitats]. AB - OBJECTIVE: To study the variation of shape of Fructus Cnidii from different habitats. METHODS: The methods of computer image processing and cluster analysis were adopted. RESULTS: The shape of Fructus Cnidii from different habitats varied significantly. CONCLUSION: According to the results of computer image processing and cluster analysis, Fructus Cnidii can be classified into three types. PMID- 14528691 TI - [Studies on fingerprint spectrum of Ephedra sinica from Gansu province]. AB - OBJECTIVE: To set up the fingerprint spectrum of Ephedra sinica from Gansu province and its extract for controlling the quality. METHODS: HPLC with UV detector was used to analyze the patterns of Ephedra sinica from Gansu province. RESULTS: The fingerprint spectrum of Ephedra sinica from Gansu province was set up. Furthermore, the fingerprint spectrum showed an excellent correlation, and all the parameters of the main co-peaks met the requirements for State Drug Administration's technical regulations on the herb. CONCLUSION: The studies are useful for the quality control of Ephedra sinica from Gansu province. PMID- 14528692 TI - [Study on effect of drug concentration of "Bushen huayu jiedu fang" drug serum on drug-resistance lung cancer cells with serum pharmacology]. AB - To explore Rh123 ion concentration effect of "Bushen huayu jiedu fang" (BSHYJD) on drug-resistance lung cancer cells, tumor-bearing animal drug-serum pharmacology and flow cytometry method were utilitied. In contrast with normal animal drug serum, BSHYJD drug-serum of tumor-bearing animal would markedly increase Rh123 concentration in drug-resistance lung cancer cells(P < 0.05), but which hasn't marked discrepancy in contrast with normal animal drug-serum(P > 0.05), that indicated BSHYJD drug-serum has increased drug concentration in drug resistance lung cancer cells. PMID- 14528693 TI - [Effects of Peristrophe roxburghiana on blood pressure. NO and ET in renal hypertensive rats]. AB - OBJECTIVE: To observe the effects of Peristrophe roxbrughiana (HSX) on blood pressure, NO, No-synthase and ET in the serum and NO in urine in renal hypertensive rats. METHODS: The 2K1C(2 kidney 1 clap) hypertensive model rats were used and drugs had been given by ig. for 4 weeks. The blood pressure was measured at the end of the each week. At the end of 4th week, the urine was collected and the concentration of NO in the serum or urine, NO-synthease and ET in the serum were measured. RESULTS: HSX (H or L dosage) by ig. for 1 week can significantly lower the blood pressure, increase the levels of NO and NO-synthase in the serum and NO in the urine, reduce the concentration of ET in the serum. The effects have lasted for 4 weeks. CONCLUSION: HSX played an important role in the renal hypertensive rats. PMID- 14528694 TI - [Blood-activating and anti-inflammatory actions of Polygala fallax]. AB - OBJECTIVE: To observe the blood-activating and anti-inflammatory actions of Polygala fallax Hesml. (PFH) in this paper. METHODS: A series of experiments were performed to show the blood-activating and anti-inflammatory actions of PFH, including acute blood-stagnation model in rats induced by dextran of high molecular mass, observation on microcirculation in mice, platelet aggregation in vitro in rabbits, acute inflammation in mice induced by xylene or histamine, and PC-DTH. RESULTS: PFH could remarkably decrease the whole blood relative viscosity. PFH could significantly dilate the micro-artery and micro-vein, increase the opening number of capillary in mice. However, PFH had no obvious effect on platelet aggregation induced by ADP in rabbits. PFH could significantly inhibit the increase of capillary permeability induced by histamine in mice, and decrease the mice ear-swelling induced by xylene. PFH could remarkably inhibit the induction-phase and attack-phase of PC-DTH in mice. CONCLUSION: The results suggested that PFH could improve the hemorrheologic character in blood-stagnation state, invigorate the micro-circulation, and inhibit inflammation process. PMID- 14528695 TI - [Antitumor effect of alcohol extract from Sesamum indicum flower on S180 and H22 experimental tumor]. AB - OBJECTIVE: To investigate the antineoplasm effect of extract from Sesamum indicum L. flower. METHOD: Observing the effects of alcohol extract from Sesamum indicum flower on tumor growth in sarcoma 180 (S180) and Heps 22 (H22) tumorigenic mouse, and on weight of immune organs. RESULT: 6, 3, 1.5 g/kg extract showed inhibiting effect on tumor growth obviously, and had not distinct effect on weight of thymus and spleen in mice sarcoma 180 and Heps 22. CONCLUSION: The alcohol extract from Sesamum indicum flower showed obvious antitumor effect. PMID- 14528696 TI - [Study on stability of beta-cyclodextrin to chlorogenic acid]. AB - To investigate the stabilizing effect of chlorogenic acid against beta cyclodextrin, beta-cyclodextrin inclusion compound of chlorogenic acid was prepared by liquid phase method, and affirmed by UV and different thermal analysis. Stability in solution was contrasted between chlorogenic acid and its inclusion complex with beta-cyclodextrin, and the stability of the latter was increased. PMID- 14528697 TI - [Study on extraction technology of soyasaponins from residual of bean ware]. AB - OBJECTIVE: To find out the optimum extraction technology of soyasaponins from residual of bean ware. METHODS: The optimum extraction conditions were investigated by the orthogonal design, and the content of soyasaponins was determined by UV-spectro-pho-tometry. RESULTS: The optimum extraction technology was A3B1C1, that is adding 7 times and 6 times amount of 70% alcohol and refluxing for two times and each time for 1.0 h. CONCLUSION: The selected technology showed higher yield of soyasaponins, good stability and high efficient. PMID- 14528698 TI - [Advances in study on the nature products of anti-myocardial ischemia]. AB - Ischemic heart disease (IHD) is a kind of severe cardiovascular disease. It has become one of the main reasons resulting in people's death. So it is very pressing to explore some active substances with anti-myocardial ischemia physiological activity. It is important to develop some safe and effective drugs. In this paper we review the advance in study on the source, chemical ingredient and pharmacological action of the anti-myocardial ischemia of thirty kinds of nature products. PMID- 14528700 TI - Complementary therapy and cancer: decision making by patients and their physicians setting a research agenda. AB - An invitational meeting, entitled Complementary and Alternative Therapy: Decision Making by Cancer Patients and Their Physicians, brought together Canadian health care providers and researchers who had expertise in patient-physician communication with those who were knowledgeable about complementary therapy and cancer. The aim was to build on the existing knowledge base in both fields in order to determine the unanswered questions, the most important questions, and what methods can be applied for answering these questions. The interdisciplinary group employed a step-wise collaborative process to develop a suggested research agenda regarding decision making by physicians and their cancer patients regarding complementary therapy. The four themes identified are establishment of a registry for complementary therapy usage for cancer care; communication; outcomes measurement; and models of integration. It is hoped that these themes will be considered worthy of support by funding agencies and worthy of investigation by researchers. PMID- 14528701 TI - Complementary therapies and cancer care: an overview. AB - The use of complementary therapies by patients with cancer is increasingly common, despite limited evidence for their efficacy and safety. The widespread use of complementary therapies has major implications for research and practice. In this paper, we provide an overview of the current state of knowledge regarding issues related to the use of complementary therapies by patients with cancer. So far, complementary therapies have not been defined very well, which makes it difficult to assess the precise extent of their use. The difference in philosophy underlying conventional and complementary treatments appears to contribute to the attractiveness of complementary therapies. Conventional medical practitioners are beginning to recognize the importance of many of the elements of this philosophy, which is evidenced in the patient-centered model. Explaining how patients with cancer make the decision to use complementary therapies has been the subject of many studies. However, as yet, no formal theory of decision making has been developed. Communication between patients and physicians about complementary therapies is an important part of the patients' decision-making process. Many gaps can be identified in knowledge of complementary therapy use by patients with cancer. Not only is there a need to increase knowledge by conducting more research and improving the research infrastructure, but attention should also be paid to information dissemination, education, and planning and development of health policies. PMID- 14528702 TI - Complementary therapies: evaluating their effectiveness in cancer. AB - The use of complementary therapies is common among cancer patients. However, a major concern is that very few of these therapies have been appropriately evaluated and, thus, little is known about their safety and efficacy. The gold standard for evaluating cancer treatments is the randomized controlled trial (RCT). However, there are several issues inherent to the nature and practice of complementary therapies that interfere with the straightforward use of RCTs. Alternative approaches are often highly individualized and attempt to respond to patients' needs. They are often holistic, taking into account many facets of a patient's life. Placebo effects and the role of the provider are frequently recognized as an important part of treatment. Outcomes of complementary therapies are often subjective, rather than being more objective outcomes, such as increased survival time. Although it is important to evaluate complementary therapies, it is mandatory that studies be sensitive to these issues and that existing research methods be adjusted and modified for this purpose. PMID- 14528703 TI - Counseling cancer patients about herbal medicine. AB - More than half of all cancer patients now use some form of complementary/alternative medicine, yet the majority of these patients do not disclose this use to their physicians. Health care practitioners need to educate themselves about the complementary/alternative medicine products their patients are using. Eight herbal products (astragalus, essiac, Asian ginseng, Siberian ginseng, green tea, garlic, Hoxsey formula and iscador) commonly used by cancer patients are reviewed here and a list of recommended reference texts is provided. In addition, health care providers are encouraged to initiate discussions about complementary/alternative products and therapies with their patients so that they may help them make safe and informed decisions about these products. Not knowing what patients are taking is definitely a less desirable option. PMID- 14528704 TI - Using the patient-centered method to achieve excellence in care for women with breast cancer. AB - The diagnosis of breast cancer remains one of the most threatening and emotionally debilitating diagnoses given to patients. Breast cancer elicits fears of loss of ability, control, hope, and at worst--loss of life. Patients and their families, devastated by the diagnosis of cancer, often feel unsupported or dismissed by their health care providers. Using the example of breast cancer, this paper describes how the application of the patient-centred clinical method can assist physicians in caring for patients diagnosed with breast cancer. Composite cases and quotes drawn from qualitative research studies with breast cancer patients are used to illustrate the application of the patient-centred clinical method. PMID- 14528705 TI - Decision making related to complementary therapies: a process of regaining control. AB - The process by which women with breast cancer make decisions related to using complementary therapies was investigated using grounded theory. Open-ended interviews were conducted with 16 women receiving orthodox treatment from across the breast cancer trajectory and who used a wide range of complementary therapies. Constant comparative analysis revealed that decision making regarding complementary therapies involved three interconnected sequential phases: (1) Getting something in place: covering all the bases, (2) hand-picking complementary therapies that fit: getting a personalized regimen in place and (3) living with the security of complementary therapies: fine-tuning a regimen to live with. Factors that influenced the decision-making process, both positively and negatively, were identified. Decision making related to the use of complementary therapies was perceived as a means of regaining control during experiences of uncertainty associated with breast cancer; the outcome of which was the maintenance of hope. Implications for supporting women's decision-making efforts regarding complementary therapies are presented. PMID- 14528706 TI - Beyond convention: describing complementary therapy use by women living with breast cancer. AB - Using a descriptive survey design, 52 women living with breast cancer were interviewed to explore their use of complementary therapy and the relationships between complementary therapy use and key demographic variables and health beliefs. Sixty-seven percent of the women reported complementary therapy use, with meditation/relaxation therapies, vitamins and spiritual healing being the three most frequently reported treatments. Women using complementary therapies were more likely to have completed post-secondary education than women using only conventional medical treatment (chi 2 = 7.1, P = 0.008). Preferred decisional role was found to be significantly associated with the use of complementary therapies (chi 2 = 11.7, P = 0.003); women using complementary therapies preferred a more active/collaborative role in treatment decisions than women using only conventional medical treatment. No significant associations were found between complementary therapy use and beliefs about cause of cancer, treatments, satisfaction with health care providers, and perceived quality of life. The findings point to the pervasiveness of complementary therapy use by women living with breast cancer and contradict past research which has supported a distinct demographic profile of complementary therapy users and associated belief system. PMID- 14528707 TI - Issues that influence patient/physician discussion of complementary therapies. AB - This article explores what patients expect from physicians when discussing complementary therapies. Counsellors at four cancer centres, under the auspices of the British Columbia Cancer Agency, discuss their clinical observations in working with oncology patients. They present perspectives of factors that influence the patient/physician discussion of complementary therapies. Some of the common themes include desire for collaboration and control, need for understanding treatment information, maintenance of hope, and trust of the physician. PMID- 14528708 TI - Should we refuse requests for complementary therapies? AB - Health care providers sometimes receive requests from patients or families for complementary therapies. When should they accede to such requests, and when should they refuse them? This is not a problem peculiar to complementary therapies. It arises whenever patients or families request conventional therapies which health care professionals judge problematic in the circumstances. What follows is a set of rationalized recommendations about when health care professionals should and should not honour requests for therapies, whether they be conventional or complementary. PMID- 14528709 TI - Clinical expertise important. PMID- 14528710 TI - The emerging roles of the practice nurse. PMID- 14528711 TI - [Computer tomography study of the pterygopalatine fossa and its communications]. AB - Pterygopalatine fossa represents very important topographic area, which is connected, by numerous foramina and fissures, to the other extracranial and intracranial areas. This gives to it huge anatomic and clinical significance. In these papers, through the method of computed tomography in axial and coronal projections, we have presented pterygopalatine fossa and the communications that it can establish through foramen rotundum (the middle cranial fossa), inferior orbital fissure (orbital cavity), sphenopalatine foramen (nasal cavity), greater palatine foramen and lesser palatine foramina (oral cavity), canalis pterygoideusa (nasal part of throat) and pterygopalatine fissure (infratemporal fossa). Knowledge on pterygopalatine fossa, and its communications, too, is necessary for adequate evaluation of the infective and tumourous processes, by which it might be primarily or secondarily taken. PMID- 14528712 TI - [Rehabilitation of war injuries of the upper extremities with peripheral nerve lesions at the and Praxis Clinic for physical medicine and rehabilitation in Sarajevo]. AB - The war injuries are the most frequent multiple, with the difficult distructions of the tissue and the lesions of the peripheral nerves. By the injuries, the lesions of the nerv system represent the delicate problem for the physical medicine and the rehabilitation during the siege of Sarajevo (1992-1995), in the injuries with the lesions of the peripheral nerves, if they are not treated in the frame of the multiple las urgent, they are postponed the operative treatments longer than three months. This is from the aspect of the successfullness, prognostically and therapeutically, the limiting moment. The successfullness of the operative treatment of the lesions of the peripheral nerves significantly depends on that whether the operation was performed incide tree months. The more difficult consequences in these injuries can appear if simultaneously with the taking care of war injuries does not perform adequately surgical treatment adn the in time physical therapy trough the sufficient long time period. On the four year sample was analyzed the success fullness of the rehabilitation of the injuries of the upper extremities with the lesions of the peripheral nerves. Clinically, uniform, we valorized the success fullness of the treatment of all the patients by the marks from 0.5. In the complete sample the excellent success of the rehabilitation we confirmed in more than a half of patients. The better results show the group of the operated patients, in which is performed neuropathia or neurolisis. The capability of the regeneration of the injured peripheral nerves offers the real possibility for extraordinary recovery also in the most difficult injuries, then the preventions of the significant number of the consequences. PMID- 14528713 TI - [Bile duct injuries in laparoscopic cholecystectomy]. AB - The aim this study was to analyse our management bile duct injuries following laparoscopic cholecystectomy. PATIENTS AND METHODS: This prospective retrospective study presents six cases of major bile duct injuries that occurred in our first 1000 laparoscopic cholecystectomies. Neuhaus et al. classification was used in this study. RESULTS: Bile duct injuries following laparoscopic cholecystectomy were identified at 6 patients, on 2 patients the lesions were identified during laparoscopic cholecystectomy. Four types of lesions were identified: 2 patients with peripheral bile leak from the cystic duct (A2), 1 patient with the complete occlusionn of the common bile duct (B2), 1 patient with lateral injury of the common bile duct(C2) and one with the right hepatic duct injury (C1), 1 patient with transection of the common bile duct (D1). A total of 6 patients with biliary injuries following laparoscopic cholecystectomy were treated at our institution: a hepaticojejunostomy was performed in 2 patients, the sutures with the use of a T-tube was performed in 2 patients, occlusion the cystic duct was performed in 2 patients. One patient after primary hepaticojejunostomy required secondary hepaticojejunostomy because of the late stricture of the anastomosis. At this writing, all the patients are well and without problems after 1 to 4 years of follow-up evaluation. CONCLUSION: The small lateral injuries of the common bile duct and peripheral leakages can be treated endoscopically, the extended lateral injuries and complete bile duct transection require surgical repair. The treatment plan must be individualized for every patient, depending on the injury type and condition of the patient. PMID- 14528714 TI - [Differences in the postoperative course and treatment in patients after laparoscopic and standard cholecystectomy]. AB - The laparoscopic surgical technique differs significantly from the classical operative technique especially concerning the post-operative treatment of the patient. Introducing a new operative technique called for an adjustment of the patients' treatment on the ward. The laparoscopic operative technique has a number of advantages in comparison with the classical operative technique. The main advantage is less damage done to the tissue, which reduces the possibility of post-operative complications. Furthermore, the rehabilitation of the patient is faster. Patients experience far less pain and the recovery period is shorter. Consecutively, this means fewer days in hospital, fewer days on sick leave, a faster recovery and returning to work sooner. Patients operated on using the laparoscopic operative technique are mobilised sooner, they start their nutrition sooner, the set of laboratory tests that are conducted is different, the time of post-operative stay at the clinic is shorter. In this paper we have analysed only operations of the holecyst over a period of one year, comparing two different operative methods. Our objective was to compare the time of post-operative mobilisation of the patient as well as the time when they start taking in liquids and food. Especially emphasised in the paper are operations with complications that call for a different treatment. In the period under analysis we did 728 holecystectomies, of which 114 were done laparoscopically. We paid special attention to the analysis of data concerning the post-operative mobilisation of the patient and the start of nutrition. The results we obtained testify in favour of the significantly shorter post-operative period without peroral nutrition following a laparoscopic holecystectomy. The period when the patient is mobilised is also different and should be given special attention. Based on this, we can conclude that at the Abdominal Surgery Clinic in Sarajevo, over the course of one year, the period of post-operative mobilisation of the patient after laparoscopic holecyctectomy has been shorter in comparison with the classical operative method. PMID- 14528715 TI - [Induced abortion: pro and con]. AB - Induced abortion like a method of birth control is the most unpopular method but it is a choice of great deal women especially in our environment. In connection with very loud demands for sharpened the low of pregnancy interrupting, many authors analyse methods, complications and risk groups of women, its acceptability like a method of family planning. At the end they give conclusion with some concrete suggestions and the aim to reduce the number of induced abortions. PMID- 14528716 TI - [Ischemic insult in the anterior and posterior cerebral circulation]. AB - In the everyday practice among clinical and etiological classifications for ischemic stroke, the terms strokes in the anterior and posterior cerebral circulation are also in use. The aim of this study was to analyze the frequency of ischemic strokes in the anterior and posterior circulation, their age and sex distribution, risk factors and hospital mortality. In the study it was analyzed 1772 patients with acute ischemic stroke hospitalized at the Department of Neurology Tuzla, Bosnia and Herzegovina, between January 1st 1996 and December 31st 2000. The mean age was 65.5 years (+9.9), 942 (55%) were females. Ischemic strokes for all patients were confirmed with computed tomography, while other data were collected from the standard patients' history charts. Anterior circulation stroke (ACS) had 1408 patients (81.8%), the rest of 314 (18.2%) had posterior circulation stroke (PCS). In the both types females were slightly overrepresented: 784 (56%) in ACS, and 158 (50.5%) in PCS. Moreover, females were significantly older than males: 67 (+9.8) versus 64 (+10) years in ACS (p < 0.001), 67.5 (+10.3) versus 63.5 (+9.2) in PCS (p < 0.001). Hypertension was the major risk factor occurring in 67% patients with ACS and 71 with PCS; heart diseases 54% in the both types, and diabetes in 23% patients with ACS and 20% with PCS. The cortical ischemic lesion was verified in 46% patients with ACS, 41% with PCS; subcortical in 12.5% and 14.5%; and lacunar in 41.5% and 44.5%, respectively. Hospital mortality was 30% (430 patients) for ACS, and 32% (101 patients) for PCS. Hospital mortality was considerably higher among females: 33% versus 28% for ACS (p = 0.03), 38% versus 27% for PCS (p = 0.03). On the basis of our study we can conclude that ischemic strokes in the anterior cerebral circulation are 4/5 of all ischemic strokes at the Department of Neurology Tuzla. Both, anterior and posterior circulation strokes are more frequent in females, witches were in average older than males. The cortical lesion dominated in ACS, and lacunar in PCS. The distribution of risk factors (hypertension, heart diseases, and diabetes) is similar for ACS and PCS, and hospital mortality also; however, mortality rate is higher in females. PMID- 14528717 TI - [Perinatal mortality at the Gynecology-Obstetrical Clinic of the Clinical Center in Sarajevo in 2001]. AB - Work results of the gynecology-obstetrics clinic Sarajevo maternity ward have been shown. Number of deliveries reached 3289, which iz significantly less then in the year 200. Prematurity reached 6.6%. Cesarian Sections were carried out in 19.3% cases and this represents a remarkable increase in comparison with the previous year 2000, when it reached 16.8%. Perinatal mortality iz 11.1@1000. Perinatal mortality shows a decrease in comparison with the period before the war. Late fetal mortality has not been significantly changed. Neonatal mortality shows an apparent decrease. Total perinatal mortality of mature newborns iz 6.9@1000 and has a constant tendency to decrease, reaching 10.5@1000 in 1991. Clinical presentation of deceased newborns predominantly presents with respiratory distress syndrome. PMID- 14528718 TI - [Ultrasound in the diagnosis and differential diagnosis of acute appendicitis in childhood]. AB - Acute appendicitis represents a leading cause of an abdominal pain in children which requires urgent surgical treatment. AIM OF THE STUDY: To investigate into value of ultrasound (US) in diagnostics and differential-diagnostics of acute appendicitis in children. PATIENTS AND METHODS: A prospective study during a two year period, was conducted to analyse abdominal US findings in 95 patients, 3-16 years of age who had acute abdominal pain. In all patients we were checking for clinical symptoms of an acute abdomen: pain, nausea, vomiting, fever, frequent urination, diarrhea, duration of symptoms and count of leukocytes. US findings were compared with pathohistological results We have used Toshiba Sonolayer SAL 77 ultrasound machine with a convex 3.75 MHz probe and a linear 8 MHz probe. RESULTS: In 50 patients sonographic diagnoses were confirmed by surgery .36 patients had other pathological conditions that lead to an acute abdomen. Nine findings were false-negative. The study results reveal that the US scanning enables not only reliable diagnosis of acute appendicitis, but also reliable differential-diagnosis of other pathological conditions and diseases. CONCLUSION: US should be the imaging method of choice in diagnostic evaluation of little patients with acute abdominal pain. This method represents a great help in differential diagnosis not only to eliminate acute appendicitis but also to confirm accuracy of the diagnosis. PMID- 14528719 TI - Suicidal thoughts related to psychoactive substance abuse among adolescents. AB - The enquiry of suicide is of particular importance. According to WHO statistics suicide in people aged from 15 to 24 years has shown the greatest increase of relative mortality. In the group of suicidal adolescents, depression, behavioural disorders, abuse of different substances and personality deviations are frequent factors in presence. The aim of our investigation was to determine the correlation between psychoactive substance abuse and the occurrence of suicidal thoughts in adolescents. The specific "Q 2000" test was used to question 600 adolescents, from Tuzla and Sarajevo Cantons, with the same gender and age distribution and rural-urban ambient. The more prevailing suicidal idea occurrence was observed in cannabis abusers (50.0%) and alcohol abusers (36.6%) in comparison to non-abusers regardless gender and/or living ambient (settlement types) (19.5% and 17.6% per each, x2 = 17.184 p = 0.00001). The augmentation in number of suicidal ideas was not observed in tobacco smokers. PMID- 14528720 TI - Types of transference studied in group of schizophrenic patients. AB - In order to find out what within the patient-therapist relationship can influence outcome, transference- countratransference patterns have been studied in the framework of psychodynamic treatment setting. The sample consisted of 40 female schizophrenic patients. All patients were interviewed by SADS-L, and diagnosed according to DSM III R rules including the course of illness. Level of social functioning was assessed on SADS-L scale. The transference/countratransference pattern was assessed by independent observer on the basis of especially designed questionnaire in active phase of illness and in the remission phase. The study found out that some transference patterns are associated with certain phase of illness, course of illness and level of social functioning. Neurotic transference pattern was associated with course of illness-remission. We do not know to which extent it is, or is it at all, related with schizophrenic's premorbid functioning or to the schizophrenic disease itself. PMID- 14528721 TI - [Chronic occupational mercury exposure in renal damage in workers in the chlorine alkali electrolysis industry]. AB - The authors investigated renal damage in 46 chlorine-alkaly plant workers (mean age was 38.8 +/- 5.7 years) under conditions of continued occupational exposure to metallic mercury vapour. The mercury unexposed control group consisted of 32 workers who works in the plant area. Significantly low of serum globulin level was found in exposed evaluated group compared with control subjects (P < 0.001). The serum globulin level was in correlation with urine mercury level (P < 0.001). Analyses of urine chemistry indicated that exposed workers had cell death produces in sediment urine as the most common signs (P < 0.001). The proteinuria was found in 4 out 32 and high level of gamma-glutamyl-transpeptidase in 8 out 32 exposed workers to high mercury level workers. Additionally, disuria and ejaculatory pain as symptoms occurred without evidence of urological disease. Mercury induced nephropathy usually associated with proteinuria, but is not with renal insufficiency. PMID- 14528722 TI - [Dental foci as important factors in the etiology of iridocyclitis]. AB - Dentodenic focuses participate in huge percentage in etiology of iridocyclitis. Beside other causes which have this eye illness like complication, such as sinusitis, diabetes mellitus, rheumatic disease, most mentioned pathological professes are these. In ophthalmology cabinet in Fojnica, during year 2002, 127 patients with dyagnosis of iridocyclitis were examined and treated, aged 9-71 both sex. After sitting of diagnosis of iridocyclitis, we want further insetting of etiological diagnosis. After complete diagnostic searches dentogenic focus was cause for this eye illness at 49 patients. After stomatological therapies such as extraction of tooth and/or conservative treatment, after short exacerbation of symptoms, immediate improvement was indicated and normalization of eye was visible, of course with adequate ophthalmological therapy. PMID- 14528723 TI - [Retroperitoneal ganglioneurinoma]. AB - We present a case of 20-years-old female with nonspecific symptoms (intermittent abdominal pains) and blood and urine tests showed the changes connected with infection of the urinal tract. The patient was sent to further diagnostic imaging methods, because the repeated attacks of uroinfection after the applied antibiotic therapy. After the US we performed CT and MRI that indicated the existence of retroperitoneal mass which dislocated the surrounding structures (left kidney and the urether) but was not infiltrated them. US were initial method and CT and MRI have done as complementary methods providing the exact picture of the mass. After surgical treatment, ganglioneurinoma was histological diagnosed. PMID- 14528724 TI - [Intracoronary radiotherapy]. AB - Intervenient cardiology is significantly more represented in attempts to solve problems of heart coronary artery disease in comparison to aortocoronary bypass procedures. Resthenoses of coronary artery blood vessel after PTCA, or PTCA and stent placement are frequent. The main causes are: neointimal hyperplasia, late contraction, and remodelling. The application of brachitherapy offers the possibility to prevent resthenosis especially stent resthenosis. Our report encompasses eight patients diseased from coronary artery occlusionn. Half of there patients had a stent resthenosis. The average degree of blood vessel occlusionn before the intervenient manipulation was 78%, and after treatment 25% 30%. Early results after recanalisation of coronary blood vessel are thus fat satisfactory, but results are to be followed carefully. PMID- 14528725 TI - [Effects of hip arthroplasty followed by by inpatient rehabilitation on physical function and quality of life]. AB - The aim of the present study was to investigate the effects of hip arthroplasty followed by an inpatient rehabilitation. Moreover, the relationships among functional status, quality of life and satisfaction with life or health status were examined. Patients were assessed before hip arthroplasty, at the start and at the end of the inpatient rehabilitation. Functional status was measured by using the WOMAC questionnaire and quality of life with the Medical Outcome Survey Short Form 36 (SF-36). Satisfaction was determined with a specific questionnaire (FLZ). Hip arthroplasty followed by an inpatient rehabilitation resulted in significant and clinically highly important improvements of functional status, quality of life and satisfaction with health and life. PMID- 14528726 TI - [Wegener's granulomatosis -- clinical presentation and diagnosis in the elderly: 6 case reports]. AB - We describe four females and two males between 63 and 81 years old, who have been treated at our department for Wegener Granulomatosis. Thereby we focus on the clinical presentation and radiological signs, and respectively to our speciality we highlight the pulmonary aspects of the disease. Referring to these six case reports we discuss the diagnosis and the clinical course of Wegener Granulomatosis in the elderly. PMID- 14528727 TI - [Massive unilateral hydrothorax as clinical manifestation of decompensated liver cirrhosis without ascites]. AB - The case of a 56-year-old man with a massive unilateral rightsided pleural effusion is described. The origin of the pleural effusion is a decompensated liver cirrhosis without ascites. The pathogenetic mechanisms and treatment are discussed. PMID- 14528728 TI - [An 80-year-old patient with known polymyalgia, worsening of general condition and weight loss]. PMID- 14528729 TI - [No cold coffee--medical effects of coffee as hotly discussed formerly as today]. PMID- 14528730 TI - Oklahoma legislature faces budget crisis affecting health care. PMID- 14528731 TI - Legislature Approves Tort Reform Measure. PMID- 14528732 TI - Barriers to information technology in healthcare. PMID- 14528733 TI - Fitch ratings. Health care special report. AB - Fitch Ratings continues to believe that the current and projected personnel shortages, especially among registered nurses (RNs). present one of the greatest operating challenges for health care providers nationwide. As noted in Fitch Research on "Health Care Staffing Shortage," dated June 21, 2001 (available on Fitch's web site at www.fitchratings. com), staffing is an area that is creating enormous expense inflation for health care providers and presenting one of the biggest areas of uncertainty in assessing an organization's credit quality. Shortages are becoming increasingly widespread, affecting health care providers in urban and rural areas regardless of size. Since health care is a labor intensive business, salary and benefit costs are key determinants of profitability. Fitch believes the attention given to the nursing shortage and potential looming crisis has created a sense of urgency, with many hospitals developing innovative strategies for work force development. In addition, federal, state, and local governments are creating programs and/or providing financial support to help alleviate shortages. Yet, the benefits of many of these initiatives may not be realized for many years, and Fitch expects health care providers to continue experiencing inflating salary and benefit expenses with growing temporary staffing usage and competitive pressure to increase overall compensation. Fitch believes managing labor costs is critical to achieving profitability, especially as the ability to increase revenue diminishes. Fitch believes the nursing shortage will continue to represent an area of extreme expense pressure for health care providers for years to come. Imbalanced supply and demand, which is expected to worsen as retiring nurses outstrip replacements, should create financial strain for many providers in an industry that is gearing up to meet the aging population's anticipated demand for more services. Fitch plans to closely monitor management strategies for dealing with vacancy and turnover rates, as well as trends in agency spending, to determine financial implications among its rated portfolio. PMID- 14528734 TI - Why nurses leave nursing: another viewpoint. PMID- 14528735 TI - New pain standards changing patterns of care in the U.S. PMID- 14528737 TI - The accelerated baccalaureate option for nursing education. Shortages, strategies, and solutions. PMID- 14528738 TI - Associate degree nursing programs. Response to the nursing shortage. PMID- 14528739 TI - Workforce diversity in hospitals. AB - With an increasingly diverse population and staff, leaders of organizations must play an active role in diversity management. Nurse leaders (n = 231) responded to a survey concerning diversity initiatives, outcomes, and issues related to staff and patients. The findings suggest that most respondents have not made diversity management an important issue. Few have goals related to diversity with indices to measure outcomes. The majority of the respondents did not report that diversity in their staffs or patients resulted in management or patient care issues. The authors suggest that managers revisit the policies, procedures, and practices related to their diversity management initiatives. PMID- 14528740 TI - Ethics and human rights issues experienced by nurses in leadership roles. AB - The aims of this study were to identify (1) the ethics and human rights issues experienced by nurses in leadership roles (NLs); (2) how frequently these issue occurred in the NLs'practices; and (3) how disturbed the NLs were by the issues. Dillman's Total Design Method (1978) for mailed surveys guided the study design. Data analysis was performed on 470 questionnaires from New England RNs in nursing leadership roles. The most frequently experienced ethics and human rights issues during the previous 12 months were (1) protecting patient right and human dignity; (2) respecting or not respecting informed consent to treatment; (3) use or nonuse of physical or chemical restraints; (4) providing care with possible risks to the RN's health; (5) following or not following advance directives; and (6) staffing patterns that limit patient access to nursing care. The most disturbing ethics and human rights issues experienced by the NLs were staffing patterns that limited patient access to nursing care, prolonging the dying process with inappropriate measures, working with unethical, incompetent, or impaired colleagues, implementing managed care policies that threaten quality of care, not considering quality of the patient's life, and caring for patients and families who are uninformed or misinformed about treatment, prognosis, or medical alternatives. Nearly 39% of the NLs reported experiencing ethics and human rights issues one to four times a week or more, and more than 90% handled their most recent ethics issue by discussing it with nursing peers. Study findings have implications for ethics education and resource support for nurses in leadership roles, and for further research on how NLs handle ethics and human rights issues in the workplace. PMID- 14528741 TI - A qualitative study of barriers and facilitators of the charge nurse role. AB - This qualitative study examines the barriers and facilitators to the role of charge nurse at a medical center. The authors interviewed 42 nursing personnel, including charge nurses (12), head nurses (10), staff nurses (11), and supervisory personnel (9). A total of 24 barriers and facilitators in three categories were identified: personal (11), interpersonal (8), and organizational (5). These factors could be used to inform developmental coaching and educational programs for charge nurses. PMID- 14528743 TI - Priorities for sleep research for older adults. PMID- 14528744 TI - The history of advance directives. A literature review. PMID- 14528742 TI - An interview with Mary Elizabeth Carnegie. Interview by Sandra Beth Lewenson. PMID- 14528745 TI - Smoking and aging. Clinical implications. Part I: Health and consequence. AB - Smoking is the number one preventable cause of disability and mortality in older adults. In the past few decades there has been an enhanced focus on smoking behaviors and smoking cessation, however, the older smoker has been excluded. Even though nonsmoking status can provide older smokers with a chance for increased quality as well as quantity of life, they are asked less often to quit, given fewer resources, and provided less guidance than younger smokers. There is limited knowledge about how to design and deliver interventions for smoking cessation among older adults. The explanation for the absence of an empirical and clinical focus on smoking cessation for the older adult is in part because of the belief in myths rather than reality. These beliefs about the health consequences of smoking behaviors and the benefits of smoking cessation for older smokers are held by older adults and their health care providers. However, the truth is that older adults can stop smoking, and the benefits are vast. PMID- 14528747 TI - Nursing needs of hospitalized older adults. Consumer and nurse perceptions. AB - The proportion of older adults is increasing in Australia, and the proportion of older adults requiring medical care is expected to increase in the future. At the same time, budget restrictions are a reality for Australia's health system. Increasing need and decreasing resources suggest the need to focus on the quality aspect of treatment and care for older adults. Little research has been conducted in the area of perceived nursing needs of elderly patients during hospitalization. This is an important area of research because it is increasingly recognized that elderly patients have specialized needs and are the major consumers of health care. Even less research has compared patient and carer perceptions with those of nursing staff. This article is a literature review and an investigation of the quality of care elderly patients receive, and of patient and nurse perceptions of the importance of various nursing activities. Quality of care is reviewed in terms of perceptions of nursing care priorities and elderly patients' satisfaction with the quality of nursing care they receive. Research examining nurses' perceptions related to why they are unable to consistently provide quality nursing care to all elderly patients is also reviewed. By identifying the nursing needs of elderly patients and educating nursing staff about these needs, professional practice can be guided and improvements in quality of care, patient satisfaction, and patient outcomes may occur. PMID- 14528746 TI - Exercise training program for older adults. Incentives and disincentives for participation. AB - The purpose of this study was to describe the incentives and disincentives for participating in an exercise training program for community-dwelling individuals 78 and older. One hundred forty-seven women and 62 men participated in this descriptive study, and ages ranged from 78 to 95 years (mean 82.7 +/- 4.1). Individuals were interviewed by phone about their decision to participate in an exercise program. Qualitative responses were analyzed using content analysis. Three main incentive themes emerged: Health Benefits, Psychological Benefits, and Positive Program Features. Disincentive themes included Competing Commitments, Doubts about Exercise, and Negative Program Features. To better tailor an exercise program to the needs of this older age group, nurses working with older adults should keep in mind the lifestyle, age-specific obligations, and set of values affecting this cohort's participation. PMID- 14528748 TI - Exploring professional caregivers' perceptions. Balancing self-care with care for patients with Alzheimer's disease. AB - With the are hundreds of studies about caregiver burden related to family caregivers that exist, little has been written about caregiver burden as it pertains to professional caregivers. The purpose of this study was to explore professional caregivers' perception and meaning associated with their caring for patients with Alzheimer's disease (AD). Twenty-two professional caregivers were interviewed. Content analysis was used to clarify respondents' interview data. Several themes emerged from the data related to beliefs about choosing and maintaining AD caregiving role, beliefs about self-efficacy, commitment and self satisfaction, nature of family interaction of individuals with AD, grieving responses, and ability to maintain self-care through social support as well as solitary time. Implications include AD professional burden measurement; study related to recruitment, orientation, and staffing patterns; and effect on the quality of care provided to patients and their families. PMID- 14528749 TI - The research process. Using a "recipe" to increase successful outcomes. PMID- 14528750 TI - NT survey highlights equipment shortages. PMID- 14528751 TI - Should school nurses take the lead in sex education? PMID- 14528752 TI - Equip us to care. PMID- 14528753 TI - The pros and cons of hormone replacement therapy. AB - Hormone replacement therapy (HRT) is used by over one million women in the UK. It was introduced 70 years ago and during this period many studies have produced evidence on both its harmful and beneficial effects. This article explains HRT and discusses the variety of other preparations available to control the effects of the menopause. The risks and benefits of HRT are gathered from current evidence. PMID- 14528754 TI - What you need to know about ... schizophrenia. PMID- 14528755 TI - Barium enema. PMID- 14528756 TI - The causes and treatments of phantom limb pain. AB - Almost all patients who undergo amputation suffer 'phantom sensations', a sensory perception of the missing limb, possibly caused by a neural imprint or memory of the limb within the brain. The patient may experience abnormal kinaesthetic sensation, such as the feeling that the limb is in an abnormal position. Perceived changes in length, size or temperature of the limb are also common. Some of these people will go on to develop phantom limb pain (PLP). Appropriate aggressive pain management is required immediately postamputation in an attempt to avoid chronic PLP. PMID- 14528757 TI - Brief intervention: reducing the repetition of deliberate self-harm. AB - Deliberate self-poisoning is one of the commonest reasons for hospital admission in the UK and people who deliberately self-harm are often difficult to engage. Solution-focused brief therapy could help these people as it has the potential to be effective after one session. The liaison psychiatry team at St Luke's Hospital, Middlesbrough, undertook a research project to evaluate the use of SFBT with patients who presented following an act of deliberate self-harm. Of the 40 patients seen only one repeated self-harm within the week of presentation. PMID- 14528758 TI - 'Volunteering is a great way to learn'. PMID- 14528759 TI - The gift of life after death. PMID- 14528760 TI - Screening to reduce HIV transmission from mother to baby. AB - From a public health perspective, targeting pregnant women for screening provides an ideal opportunity to prevent the spread of human immunodeficiency virus (HIV). Midwives are ideally placed to recommend and offer antenatal screening as part of the National Universal HIV Screening Policy (Department of Health, 1999). The rationale behind antenatal screening is to reduce transmission of HIV from an infected mother to her baby, a process that can occur during pregnancy, labour and through breastfeeding. PMID- 14528761 TI - What is genital chlamydia? AB - Genital chlamydia is a sexually transmitted infection (STI) caused by the intracellular bacterium Chlamydia trachomatis. Infection occurs during vaginal, oral or anal sex, or other genital contact with a partner who has chlamydia. Newborn babies can also acquire the infection during birth. This can lead to conjunctivitis or severe chest infection. PMID- 14528762 TI - The nurse role in implementing the national sexual health strategy. AB - The rising incidence of sexually transmitted infections (STIs) in England and a recognition that looking after our sexual health is essential for long-term well being, prompted the government to unveil its first sexual health and HIV strategy (Department of Health, 2001). The government's aim is to modernize sexual health and HIV services and help primary care trusts (PCTs) to address the rise in STIs and HIV, and to reduce unintended pregnancies. PMID- 14528763 TI - The sexual health needs of people with learning disabilities. AB - The sexual health of people with learning disabilities raises important management and practice issues for health services, and should be examined in the context of the current policy emphasis on advocacy, person-centred services and social inclusion (Department of Health, 2001). People with learning disabilities may have limited access to mainstream health services, and sexual health and genitourinary medicine (GUM) services are no exception (DoH, 2001; 1998). They are often excluded from society, either because they are 'segregated' within specialist support services in the community or because they live in isolation with carers, and health and social care models do not always join up locally to meet their needs. PMID- 14528764 TI - How to ... interview effectively. PMID- 14528765 TI - Evolution or extinction? Which will we choose? PMID- 14528766 TI - Do your pediatric patients have chronic health conditions? Medical history responses may not be accurate. AB - A review was conducted of a series of studies that raised questions regarding the accuracy of: 1) reports on the national prevalence rates of chronic disabling conditions in children, and 2) parental reports to individual practitioners on the health condition of their children. Practitioners are alerted to the seriousness of these realities. PMID- 14528767 TI - Behavioral evaluation during dental care in children ages 0 to 3 years. AB - PURPOSE: Many studies concluded that the behavior of babies complicated dental care because of child immaturity or the difficulty of establishing acceptable communication and comprehension. METHODS: The records of 696 babies were randomly selected from the baby clinic of Aracatuba in Brazil. Patient age ranged from 0 to 36 months. They were divided into 6 groups according to age: Group I-0 to 6 months; Group II-7 to 12 months; Group III-13 to 18 months; Group IV-19 to 24 months; Group V-25 to 30 months; Group VI-31 to 36 months. The behavior of the child was evaluated upon entrance in the dental office and during the first 4 clinical appointments with a clinical exam and oral physiotherapy. The baby was classified as collaborator (C) or noncollaborator (NC). Statistical analysis was performed using Pearson's chi-square method (P < .05). RESULTS: The percentage of NC for Groups II, III, IV, and V (66%), was significantly higher than for groups I (30%) and VI (50%). CONCLUSIONS: Babies from 0 to 6 months showed a collaborative behavior; babies from 7 to 30 months showed noncollaborative behavior; and babies from 31 to 36 months showed no statistically significant difference between the percentage of C and NC. PMID- 14528768 TI - Attitudes of Saudi parents toward behavior management techniques in pediatric dentistry. AB - PURPOSE: The purpose of this study was to investigate the attitudes of the Saudi parents toward the different behavior management techniques (BMT) used in pediatric dentistry and assess the factors affecting such attitudes. METHODS: Two videotapes, each containing 10 BMT, were produced, one with and the other without explanation and objectives of each technique. Parents (N = 133) watched the videotapes as individuals or in groups and rated the acceptability of each BMT. RESULTS: The results indicated that generally Saudi parents readily accept most BMT. Informed parents reported higher acceptance levels of BMT than the uninformed parents (F = 23.975, P = .0001). Group viewers reported less acceptance levels than individual viewers (F = 46.940, P = .001). The tell-show do, positive reinforcement, and distraction were the most readily acceptable techniques, while parent separation, physical restraint, hand over mouth, and voice control were the least accepted techniques. CONCLUSIONS: Generally Saudi parents accepted most of the BMT. Explanation enhanced their level of acceptance. PMID- 14528769 TI - Mutans streptococcus counts following treatment for early childhood caries. AB - PURPOSE: The purpose of the study was to assess mutans streptococci (MS) count in children who were treated for early childhood caries (ECC) in a follow-up examination. METHODS: Forty-four children who were treated for ECC in the pediatric dentistry clinic of the Hebrew University-Hadassah School of Dental Medicine were given paraffin film to chew. A blade containing children's saliva was placed inside a test tube containing MS selective medium to allow bacteria growth. The kits were transferred immediately to the laboratory and incubated for 48 hours in a 37 degrees C incubator. After incubation, bacterial growth was counted by the amount of colonies formed. RESULTS: Children who were examined closer to the end of the dental treatment demonstrated significantly lower MS counts. In addition, children who had higher birth weight also demonstrated lower bacterial counts. Significantly more parents of children with the high MS group than in the low MS group count reported that oral hygiene instructions greatly reduced children's plaque levels. All the children who were born preterm had high MS count, while 77% of children who were born at term demonstrated high MS count. CONCLUSIONS: Children who had dental treatment due to ECC may have high MS count in the future and are at risk for caries attack. Factors that may be associated with high MS counts are low birth weight and mother's education. PMID- 14528770 TI - Comparison of diagnostic methods for dental caries. AB - PURPOSE: The purpose of this study was to compare results of 4 different methods of diagnosing dental caries in 12-year-old children by evaluating their influence on the DMFS index and analyzing the correlation of the radiographic exam with the clinical exam. METHODS: The tools used were: 1) dental mirror; 2) dental mirror and dental explorer, both performed in an outdoor setting under natural light; 3) dental mirror; 4) dental mirror and dental explorer, both performed in a traditional dental setting under artificial light. The radiographic exam was carried out after the clinical exam. RESULTS: The dental explorer may be omitted from the clinical exam in epidemiological surveys and in clinical exams performed in dental offices. Methods 3) and 4) performed better in diagnosing dental caries than methods 1) and 2), which underestimated decay. CONCLUSIONS: In the traditional dental setting, the process of dental cleaning and drying as well as artificial light, improved the diagnosis of dental caries. The radiographic exam used for epidemiological surveys, in relation to method 2), improved the clinical exam by 4%. PMID- 14528771 TI - Influence of dental care for infants on caries prevalence: a comparative study. AB - PURPOSE: The influence of early dental care on the prevalence of dental caries was determined in children ages 35 to 40 months divided into 2 groups of 160 children each, with 1 group participating in a dental care program from the first year of life to the time of the study, while the other group did not receive any dental care. METHODS: The clinical conditions considered for the evaluation were: sound teeth, enamel caries without cavitation, enamel caries with cavitation, and dentinal caries. The proportion and chi-square tests were used for statistical analysis with the level of significance set at 5%. RESULTS: A significant difference regarding the presence of dental caries, especially in the number of children presenting enamel caries with cavitation (P < .0001), was observed between the 2 groups. The number of children with enamel caries without cavitation and dentinal caries was similar for the 2 groups. CONCLUSIONS: Considering the aspect of dental caries prevention, the authors concluded that children in the age range of 3 to 4 years who received early dental care showed better oral conditions. PMID- 14528772 TI - Caries prevalence in northwest Michigan migrant children. AB - PURPOSE: The purpose of this study was to describe the prevalence of dental caries in a sample of Hispanic-American migrant workers' children and compare the results with an earlier Northwest Michigan migrant study to assess if migrant children have benefited from the global caries decline. METHODS: A non probability convenience sample of migrant farm worker children was obtained from migrant schools in Northwest Michigan during the summer of 1999. Field dental equipment was used to examine caries, fluorosis, and urgent dental needs by 1 examiner. Statistical analysis was performed using SAS software with the student t-test used to test for differences between the samples. Intra-examiner reliability was high for caries at the surface level (kappa = 0.94). RESULTS: The sample consisted of 211 children with 95% ranging in ages from 5 to 13. Mean caries levels were as follows: DMFT = 1.1 (SD 1.6); DMFS = 1.6 (SD 2.5); dft = 3.0 (SD 2.9); dfs = 6.3 (SD 6.7) Fifty percent of children with at least 1 permanent tooth (189) were caries free, with 17% (34) having DMFT values greater than or equal to 3. The large number of caries-free children displayed a skewed distribution of caries where a small proportion of the population harbored the majority of the disease. A statistically significant decrease in caries was evident in permanent teeth (P = .0001) between 1999 and 1982 samples, but no change was realized in primary teeth (P = .7). CONCLUSIONS: Results indicated that the caries decline has extended to this long-neglected migrant group, but only in the permanent dentition. Caries remained moderate to high in the primary dentition and the caries burden was carried by a relatively small segment of the population. Dental treatment programs should be expanded to include preschool children who appear to be at the highest risk for caries. PMID- 14528774 TI - Comparative study of microleakage of a pit and fissure sealant placed after preparation by Er:YAG laser in permanent molars. AB - PURPOSE: The aim of this study was to assess the microleakage of a pit and fissure sealant on enamel treated with a laser, with and without etching. METHODS: Sixty non-carious extracted molars were randomly assigned to 2 groups. For both groups, in the mesial halves, the fissures were widened mechanically with a bur and etched for 15 seconds. In group 1, the distal half was prepared with a laser alone; in group 2, a laser was followed by etching. Then the sealant was applied on all teeth. RESULTS: Laser alone showed the highest number of specimens with microleakage (63%) (P < .02) and the highest mean of microleakage (0.76 mm vs 0.12 mm, 0.17 mm, and 0.18 mm; P < .01). CONCLUSIONS: No significant difference was noted between the 2 types of enamel preparation when etching was performed. Laser irradiation did not eliminate the need for etching the enamel surface before applying the sealant. PMID- 14528773 TI - The assessment of pain sensation during local anesthesia using a computerized local anesthesia (Wand) and a conventional syringe. AB - PURPOSE: The purpose of this study was to compare the behavior reaction of children who received local anesthesia with a conventional syringe injection and a computerized device (Wand). METHODS: One hundred and two children ages 3 to 10 years were selected for this study. In 1 group there were 55 children between the ages of 3 to 5 years old, and in the other group there were 47 children ages 6 to 10 years old. They all needed at least 2 clinical sessions of operative procedures, preceded by a local anesthetic injection, 1 on either side of the same jaw. The local anesthesia was delivered using either the Wand or the traditional syringe. A random crossover design was used so that each child served as his/her own control. RESULTS: There were 25 girls and 30 boys in group A (mean 4.1 +/- 0.6 years), and 26 boys and 21 girls in group B (mean 7.2 +/- 1.3 years). Most of the children had a good reaction to both techniques of local anesthesia. No significant difference in either group was found between boys and girls. The children's reactions to injection in the mandible or the maxilla with the Wand or the conventional technique, regarding crying, facial expression, hands, legs and torso movements were similar with no statistically significant difference. There was no statistical difference when the maxillary infiltration was delivered to 1 or multiple teeth. No significant difference was found when the Wand was delivered during the first or second visit. CONCLUSIONS: The results suggested there was no difference in the pain behavior of children during the administration of local anesthesia with a conventional injection or a computerized device when the operator was an experienced pediatric dentist. This was true for maxillary infiltration and mandibular block. For other techniques, such as palatal injection and periodontal ligament injection, more studies should be conducted. PMID- 14528775 TI - Effect of dentinal tubule orientation on the microtensile bond strength to primary dentin. AB - Direction of dentin tubules is an important factor that affects bond strength of contemporary adhesive materials. Because conservative proximal cavity preparations in primary teeth contain more dentin tubules running oblique or parallel to the cavity walls, this study was conducted to evaluate the influence of tubule orientation on the bond strength to primary dentin. A polyacid-modified resin composite material was used as the test material. A microtensile test was utilized to enable evaluation of bond strengths in three different tubule orientations (perpendicular, oblique, parallel) following aging of the resindentin bonds by long-term immersion of the test samples in water. Samples bonded parallel to the dentin tubules showed the highest microtensile bond strength, followed by the oblique and perpendicular groups. Statistical analysis of the data showed no significant difference between the bond strength of parallel and oblique groups. Independent of the tubule orientation, all samples revealed cohesive fracture within dentin. PMID- 14528776 TI - Bond strengths of Dyract AP compomer material to dentin of permanent and primary molars: phosphoric acid versus non-rinse conditioner. AB - PURPOSE: The purpose of this in vitro study was to determine the shear bond strengths (SBS) of Non-Rinse Conditioner (NRC, Dentsply) combined with Prime & Bond NT (PBNT, Dentsply), a 1-bottle adhesive. The null hypothesis tested was that the use of NRC with PBNT would not result in SBS different from those obtained with conventional phosphoric acid (PA) etching and bonding application to permanent and primary dentin. METHODS: Extracted human third molars and primary molars were mounted length-wise in acrylic resin. The occlusal surfaces were ground to expose a flat dentin surface, and then polished to 600 grit silicon carbide paper to create smear layers similar to those created with high speed burs. The teeth were randomly assigned to 4 groups (N = 10) according to the etchant/conditioner (PA vs NRC) and dentin (permanent vs primary) used: (Group I: permanent dentin, PA, PBNT; Group II: primary dentin, PA, PBNT; Group III: permanent dentin, NRC, PBNT; Group IV: primary dentin, NRC, PBNT). Specimens were then secured in a split mold, having a 5 mm diameter opening and a polyacid modified resin composite (Dyract AP, Dentsply) were inserted and light cured incrementally onto the treated dentin surfaces. All specimens were stored in water for 24 hours prior to shear strength testing using a Franell testing machine at a crosshead speed of 0.8 mm/minute. RESULTS: The mean dentin SBS values (MPa) for the groups were: Group I (13.32 +/- 6.6); Group II (15.21 +/- 5.25); Group III (8.87 +/- 3.12); and Group IV (7.42 +/- 2.98). Analysis of variance and Duncan's multiple range tests indicated significant differences among groups at P < 0.05. CONCLUSIONS: In general, the SBS were remarkably greater in the 2 groups etched with PA in comparison with the 2 groups conditioned with NRC. However, the type of dentin tissue did not influence SBS. PMID- 14528777 TI - A study on the dental disease of the handicapped. AB - Opinions vary as to what extent handicapped children differ in oral health from children without handicapping conditions. The present study was a comprehensive study and evaluation of oral health comparing 267 handicapped and 128 nonhandicapped persons with regard to caries rate, occlusion, and periodontal condition. The dft, dfs, and DMFT indices of handicapped according to age were significantly lower than those of nonhandicapped subjects. The prevalence of gingivitis in handicapped persons increased with age and was significantly higher than that reported for nonhandicapped subjects. The handicapped group had a significantly higher percentage of Class II and Class III occlusion than that observed in the comparison group. PMID- 14528778 TI - Orthodontic treatment need of mentally handicapped children in Ibadan, Nigeria, according to the dental aesthetic index. AB - PURPOSE: Proper well-organized orthodontic treatment planning for special needs patients quantifies such needs with a valid screening method to determine priority treatment. The aim of this study was to determine the orthodontic treatment needs of mentally handicapped children in Ibadan, Nigeria. METHODS: The sample consisted of 124 mentally handicapped children, 68 boys and 56 girls who were 6 to 18 years old with mean age of 14.5 +/- 1.5 years, drawn from 7 special centers/schools in the city. RESULTS: Less than 42% had normal or minor malocclusions needing slight or no treatment; 17% had definite malocclusions with treatment elective; 9% had severe malocclusions and treatment highly desirable; 32% had very severe or handicapping malocclusions with treatment considered mandatory. Significant gender differences were found in the mean dental aesthetic index (DAI) scores (P < .05) and significant association noted (P < .05) between severe mental retardation and DAI scores deserving treatment. CONCLUSIONS: There were considerable orthodontic treatment needs among the study population with a reasonable percentage deserving treatment in publicly funded programs. PMID- 14528779 TI - Dental crowding in primary dentition and its relationship to arch and crown dimensions. AB - PURPOSE: The purpose of this study was to compare dental arch size, mesiodistal and buccolingual crown widths, and crown shapes between 2 groups of children, 1 with anterior crowding in both dental arches and another with anterior spacing in both dental arches. METHODS: Sixty-one sets (27 crowded and 34 spaced arches) of dental casts of normal occlusions were examined. RESULTS: For both lower and upper arches, crowded arches had statistically significantly smaller arch widths than did the spaced arches. The buccolingual width of the upper second primary molar of the crowded arches was statistically significantly larger than that of the spaced arches. However, there were little significant differences between crowded and spaced arches in arch length, mesiodistal crown width, and crown shape. CONCLUSIONS: It can be concluded that the factor that determines whether a primary dental arch is crowded is the absolute width of the arch. PMID- 14528780 TI - Systemic manifestations during eruption of primary teeth in infants. AB - PURPOSE: The purpose of this study was to describe the most frequent objective clinical manifestations found during the eruption of primary teeth and to assess the correlation between those symptoms and the eruption of various groups of teeth. METHODS: The study was conducted in the Clinica Infantil Colsubsidio in Bogota, Colombia. Five hundred and eighty-five children participated in the study. One-hundred and forty-five children ages 4 to 36 months who had at least 1 erupting tooth were included in the study group. The other 357 children served as a control group. Parents were asked to complete a short questionnaire and children were then checked by one of the authors. Chi-square analysis was performed to analyze information obtained for the 2 groups. Level of significance was set at P < .05. RESULTS: There were 81 girls (56%) and 64 boys (44%) in the study group, and 183 girls (54%) and 157 boys (46%) in the control group. The most frequent clinical manifestations were: drooling (15%), diarrhea (13%) and drooling-diarrhea (8%). The presence of fever and fever-diarrhea was shown in a lower percentage (8%). In the study group, boys demonstrated a higher prevalence of diarrhea than girls (P < .05). No statistical significance regarding other clinical manifestations and gender were observed. In the control group, 93% of the children did not show any clinical manifestation. CONCLUSIONS: An association has been shown between general objective manifestations like drooling (the most prevalent), fever and diarrhea, and the eruption of primary teeth. Most manifestations appeared during the eruption of the primary incisors. PMID- 14528781 TI - Orthodontic treatment of a patient with an unusual impaction of a central incisor: case report. AB - This report documented an unusual case with impaction of a right permanent central incisor. The purpose of this work was to illustrate how the original lingual lever arm moved the impacted central incisor in the dental arch. The authors used a modified lingual arch made of .036 inch stainless steel wire soldered to bands on the second primary molars. The lingual arch caused some minor inconveniences, but the discomfort was slight. Patient compliance was not a problem because a fixed appliance was used in the primary dentition. PMID- 14528782 TI - Management of occlusal and developmental disturbances resulting from an ankylosed maxillary second primary molar: case report. AB - This case report presented the orthodontic management of an ankylosed primary molar. Trauma to the maxillary right second primary molar resulted in the infraocclusion of the affected tooth as well as the mesial tipping of the adjacent first permanent molar and displacement of the permanent successor. After extraction of the ankylosed primary molar, orthodontic therapy was performed to upright the adjacent permanent molar and to gain the lost space of its permanent successor. By the application of orthodontic mechanics, the maxillary right permanent first molar was uprighted and the maxillary right permanent second premolar was brought to its proper position. PMID- 14528783 TI - Stemming the tide: cognitive aging theories and their implications for continuing education in the health professions. AB - As demographic drift among health care providers mimics that of the larger population, it becomes increasingly clear that theory pertaining to the impact of aging on cognitive processing should inform the continuing education efforts designed for health care professionals. The purpose of this article is to offer a critical review of the major theories in this area and outline a sample of the implications that can be derived from these views. Research articles examining the relationship between age and physician performance were identified using MEDLINE, PsychLit, and ERIC. In addition, the psychology literature on age related changes in cognitive processing was reviewed. Evidence from the medical education literature and psychological theory suggest the importance of increased environmental supports, decreased time demands, and peer review programs as barriers against the impact of aging. The implications of these findings include the potential to tailor continuing education (and physician remediation) efforts toward the age-related abilities/deficiencies of individual physicians. PMID- 14528784 TI - Building on abilities. AB - This article reviews generalizations about effective continuing education that pertain to cognitive aging and memory and to a broader range of influences related to adult learning and development and to the context of professional practice. Features of excellent continuing education include attention to diversity, objectives, motivation, active learning, feedback, and application. The learning abilities and capacities of most professionals are sufficient for them to continue to enhance their proficiencies throughout their careers. Cognitive aging is one influence among various psychological characteristics. Effective continuing professional education should address career span learning and performance, including emphasis on abilities and not just disabilities. PMID- 14528785 TI - Impact studies in continuing education for health professions: update. AB - INTRODUCTION: This article critiques the questions asked and methods used in research syntheses in continuing education (CE) in the health professions, summarizes the findings of the syntheses, and makes recommendations for future CE research and practice. METHODS: We identified 15 research syntheses published after 1993 in which primary CE studies were reviewed and the performance (behavior) of health professionals and/or patient health outcomes were examined. RESULTS: The syntheses were categorized by the research questions they asked using a wave metaphor. Wave One (n = 3) syntheses sought to establish a general descriptive causal connection between CE and impact variables, asking, "Is CE effective, and for what outcomes?" Wave Two syntheses (n = 12) sought to explain the relationship between CE and impacts by identifying causal moderators, asking, "What kinds of CE are effective?" DISCUSSION: Wave One findings confirm previous research that CE can improve knowledge, skills, attitudes, behavior, and patient health outcomes. Wave Two syntheses show that CE, which is ongoing, interactive, contextually relevant, and based on needs assessment, can improve knowledge, skills, attitudes, behavior, and health care outcomes. The most important implication of the present review is that there are differential impacts of CE programs, and the reasons for those impacts cannot be fully understood unless the context of the program is considered. PMID- 14528786 TI - Using a quasi-experimental research design to assess knowledge in continuing medical education programs. AB - INTRODUCTION: The objectives of continuing medical education (CME) programs include knowledge acquisition, skill development, clinical reasoning and decision making, and health care outcomes. We conducted a year-long medical education research study in which knowledge acquisition in our CME programs was assessed. METHOD: A randomized separate-sample pretest/past-test design, a quasi experimental technique, was used. Nine CME programs with a sufficient number of participants were identified a priori. Knowledge acquisition was compared between the control group and the intervention group for the nine individual programs and for the combined programs. RESULTS: A total of 667 physicians, nurses, and other health professionals participated. Significant gain in knowledge was found for six programs: Perinatology, Pain Management, Fertility Care 2, Pediatrics, Colorectal Diseases, and Alzheimer's Disease (each p < .001). Also, the intervention group differed from the control group when the nine programs were combined (p < .001), with an effect size of .84. DISCUSSION: The use of sound quasi-experimental research methodology (separate-sample pretest/post-test design), the inclusion of a representative sample of CME programs, and the analysis of nearly 700 subjects led us to have confidence in concluding that our CME participants acquired a meaningful amount of new knowledge. PMID- 14528787 TI - Continuing medical education: what delivery format do physicians prefer? AB - BACKGROUND: Although physicians are in a unique position to prevent life threatening outcomes by counseling patients to stop smoking, many of them miss the opportunity to intervene in their patients' use of tobacco. Nicotine Dependence Across the Lifespan was developed as a continuing medical education (CME) program to teach and encourage physicians to deliver effective tobacco prevention and tobacco cessation counseling. METHODS: This CME program was offered to Texas physicians, free of charge, in three formats: live lectures, videotapes, and World Wide Web-based training. The program targeted physicians in four rural areas of Texas (San Angelo, Harlingen, Tyler, and Lubbock), where high smoking rates are prevalent and the number of professional smoking cessation services is low. We examined the sociodemographic characteristics of the participating physicians, factors in their decisions to participate in the program, and the extent to which their reported CME format preferences were associated with age, gender, race, profession, and location. RESULTS: The four factors identified--professional development, cost, personal control, and convenience/complexity--explained 76.9% of the variance describing the reasons physicians participated in the CME offering. The physicians' preferred CME format was live lectures; based on responses, this did not differ across age, gender, race/ethnicity, and location. DISCUSSION: Live lecture continues to be a preferred format of CME for physicians in four rural areas of Texas, yet research continues to show that lecture results in only the lowest level of behavioral change. PMID- 14528788 TI - Postgraduate educational program for primary care physicians in remote areas in Lebanon. AB - INTRODUCTION: Continuing medical education (CME) is a requirement in many developed countries. Lebanon lacks such a rule; hence, the dictum "once a doctor always a doctor" holds. This article describes a pioneering postgraduate educational program for primary care physicians in remote areas of Lebanon. METHOD: The Lebanese Society of Family Medicine introduced a 2-year structured CME program to four remote Lebanese areas. Thirteen family physicians provided 33 activities to 1,073 primary care physicians tackling 22 subjects chosen from a list of 53 common clinical problems approved by community medical leaders. Each attendee was requested to complete an evaluation form at the end of each session. RESULTS: The activities were attended by 1,073 physicians, 914 of whom filled out the evaluation forms (85.2% response rate). The analysis of the response of the attendees revealed that 65% of the attendees completely agreed that they benefited from the activities, 68% completely agreed that the presentations were clear, 86% thought that the methods used were adequate, 57% agreed completely that the presenters were prepared, and 69% replied that enough time was available for interaction. DISCUSSION: The CME programs were conducted with minimal costs. They were well received by attendees. It is recommended that the Lebanese health authorities make CME a requirement to promote the knowledge and behavior of primary care physicians and improve health. PMID- 14528789 TI - Unanticipated learning outcomes associated with commitment to change in continuing medical education. AB - INTRODUCTION: Educator-derived, predetermined instructional objectives are integral to the traditional instructional model and form the linkage between instructional design and postinstruction evaluation. The traditional model does not consider unanticipated learning outcomes. We explored the contribution of learner-identified desired outcomes compared with learner outcomes that were not named in the instructional design. METHOD: This study was conducted at a short course in pediatrics in which 43 physicians, advanced practice nurses, nurses, and physician assistants voluntarily self-identified committed- to changes (CTCs). We compared these CTC predicates with the predetermined instructional objectives that had been published in advance in the conference brochure and syllabus. CTCs whose predicates described the same features as the instructional objectives were considered to be anticipated learning outcomes. CTCs lacking correspondence with instructional objectives were considered to represent unanticipated learning outcomes. RESULTS: Of the 157 CTCs, 68% were anticipated learning outcomes because their predicates could be linked to the instructional objectives. The remaining 32% of CTCs did not correspond to any of the instructional objectives and thus represented unanticipated learning outcomes. DISCUSSION: These findings demonstrate that evaluations based on instructional objectives, although valuable, are incomplete because educational activities may also stimulate many unanticipated learning outcomes. Continuing medical education planners can gain a fuller assessment of the effect of their educational endeavors by including predetermined instructional objectives and encouraging the constructivist practice of recognizing unanticipated learning. PMID- 14528790 TI - Assessment of clinical skills in medical practice. AB - The introduction of a clinical skills examination (CSE) to Step 2 of the U.S. Medical Licensing Examination (USMLE) has focused attention on the design and delivery of large-scale standardized tests of clinical skills and raised the question of the appropriateness of evaluation of these competencies across the span of a physician's career. This initiative coincides with growing pressure to periodically assess the continued competence of physicians in practice. The USMLE CSE is designed to certify that candidates have the basic clinical skills required for the safe and effective practice of medicine in the supervised environment of postgraduate training. These include history taking, physical examination, effective communication with patients and other members of the health care team, and clear and accurate documentation of diagnostic impressions and plans for further assessment. The USMLE CSE does not assess procedural skills. As physicians progress through training and enter practice, both knowledge base and requisite technical skills become more diverse. A variety of indirect and direct measures are available for evaluating physicians, but, at present, no single method permits high-stake inferences about clinical skills. Systematic and standardized assessments make a contribution to comprehensive evaluations, but they retain an element of assessing capacity rather than authentic performance in practice. Much work is needed to identify the optimal combination of methods to be employed in support of programs to ensure maintenance of competence of practicing physicians. PMID- 14528791 TI - Quality. Pay for good play. PMID- 14528792 TI - Insurance. Comp complaints. PMID- 14528793 TI - Patient safety. X marks the spot. PMID- 14528794 TI - The data page. Medicaid cuts threaten kids coverage. PMID- 14528795 TI - Information technology. Help is on the way. PMID- 14528796 TI - Information technology. Integrating IT. PMID- 14528797 TI - Construction. Class is now in session. PMID- 14528798 TI - HIPAA. All systems go. PMID- 14528799 TI - Insurers reign. AB - Double-digit premium increases, an easing of runaway cost growth and less competition have led to record profits and climbing stock prices for the nation's insurance companies. While the immediate outlook is rosy, there are hints that a backlash may be developing among providers and consumers disgruntled over the high premiums, and among policy-makers alarmed at the climb in the number of uninsured Americans, which is estimated at 41 million now and is expected by all forecasters to continue to surge. PMID- 14528800 TI - Delicate balance: 2003 salary survey. AB - Hospital boards are exercising more scrutiny than ever on executive pay, at a time when the disparity between salaries in health care and general industry has finally disappeared. PMID- 14528801 TI - Motor city blues. AB - With its safety net hospital in peril, Detroit had a choice: fundamentally reform its public health system or watch it collapse. Cities nationwide face a similar dilemma. PMID- 14528802 TI - How health care won its first Baldrige. AB - SSM Health Care last year won the nation's most prestigious award for business quality. How it did so provides valuable lessons for all hospitals striving for excellence. PMID- 14528803 TI - Quest for quality prize. AB - Abington (Pa.) Memorial Hospital won this year's American Hospital Quest for Quality Prize for its patient safety program, which aims to head off harm when errors occur. PMID- 14528804 TI - Executive dialogue: The excellent manager. AB - In an H&HN roundtable, health care human resources executives and consultants examine characteristics of excellent managers and how to identify them in the hiring process. PMID- 14528805 TI - Standing still. Medical advances are infused into a system that has changed little since the turn of the 20th century. PMID- 14528806 TI - The engineering of an artery. AB - Tissue engineering is seen by some to be a high-risk area that has yet to deliver any clinical success. This article addresses one particular area of high risk, the tissue-engineered artery, and discusses some of technical and clinical factors that are involved. PMID- 14528807 TI - The functionality and cost advantages of high-performance polymers. AB - Acetals remain extremely important for medical devices, particularly in gears, springs and other mechanisms, although going forward with progressively lower emission targets is likely to require a combination of low-emission grades and tighter processing controls. Nylon and PBT materials have a continued importance in achieving the combination of mechanical performance, biocompatibility (a range of grades are available that have been tested successfully against USP 23 Class VI) and sterilisation performance (dependent on grade and type of sterilisation). Materials such as liquid crystal polymer are progressively more important for their barrier properties, high temperature performance and all-round sterilisation performance. Polycarbonates and cyclic olefin copolymers continue to find new applications, often where clarity is important; transparent Nylons and other olefinic materials are also valuable in this area. With the continuing advances in raw materials and polymer processes, careful choices can produce some worthy advances in device technology, although utilising the technologies effectively still depends on working forwards from the user/patient need and desired functionality. Whether considering developing a new device using plastics, or reconsidering further development of an existing device, engineering polymers can provide the key to something better. PMID- 14528808 TI - Combining local drug delivery and implantable medical devices. AB - The targeted, controlled local delivery of a drug by an implant is the optimum method of treating a range of conditions. This review of product developments discusses potential future device applications and progress with novel techniques employing drug delivery matrices and coating methods. PMID- 14528809 TI - Classification and characteristics of polymeric controlled-release systems. AB - The great importance of controlled release for the delivery of a variety of bioactive agents is evidenced by an increasing number of patents and commercial products that employ this technology. This article reviews the different controlled-release systems and their operating mechanisms. PMID- 14528810 TI - Selecting adhesives. AB - It can be seen, therefore, that attaining optimum performance and durability from a medical device is possible if the design of the joints in the device are carefully planned in conjunction with an informed selection of an adhesive. PMID- 14528811 TI - Wrist-wearable medical devices: technologies and applications. AB - Standard HDP technologies such as COB can be involved early in the development stage. They help to form the development process and to meet size requirements without exhausting the development budget. Higher HDP production costs are negligible compared with the total development, field test and regulatory approval costs. Using HDP for miniaturisation, it is possible to make applications of previously lab-scale equipment for WMDs and for implants. PMID- 14528812 TI - Managing changes during a clinical investigation, Part I. AB - During the course of a clinical investigation, changes may need to be made to the device, the manufacturing process, the investigational plan or other aspects of the investigation. Part I of this article discusses the United States requirements for managing these types of changes. Part II will discuss European requirements and the need for developing a standard operating procedure to help ensure consistent compliance. PMID- 14528813 TI - Human tissue-engineered products. AB - Exciting developments are taking place that present a complex challenge for regulations and standards. PMID- 14528814 TI - How effective is your safety culture? AB - Safety cultures are nothing new. How a company implements its management systems and determines who will drive them will ultimately affect its success. PMID- 14528815 TI - Two lessons in compliance. PMID- 14528816 TI - New lab ventilation standard. ANSI/AIHA Z9.5-2003 affects every lab. PMID- 14528817 TI - The seven potentially deadly sins of safety. PMID- 14528818 TI - Selecting for safety. PMID- 14528819 TI - Standards and trends in the glove industry. PMID- 14528820 TI - Hand protection for electrical safety. PMID- 14528821 TI - Good foot protection revolves around common sense. PMID- 14528822 TI - When a good work shoe isn't enough. PMID- 14528823 TI - Responding to 'dirty bombs'. PMID- 14528824 TI - Behind the HAZMAT regs. PMID- 14528825 TI - Building corporate castles, Part 3. PMID- 14528826 TI - Integrated safety programs: from the top down to the bottom line. PMID- 14528827 TI - Playing your cards right. PMID- 14528828 TI - In defense of incentives & recognition. PMID- 14528829 TI - Standing for comfort. PMID- 14528830 TI - Performing effective ergonomic evaluations. PMID- 14528831 TI - Wipe out slips, trips, and falls. PMID- 14528832 TI - NRR = not really relevant? PMID- 14528833 TI - Getting the most out of forklift alarms. PMID- 14528834 TI - Evaluating multimedia. PMID- 14528835 TI - New security requirements for HAZMAT transportation. PMID- 14528836 TI - Keep plugging. PMID- 14528837 TI - Occupational asthma in health care professionals. PMID- 14528838 TI - Hewlett-Packard cuts ergonomics risks. PMID- 14528840 TI - [Health-related quality of life of the Hungarian population]. AB - AIMS: The objective of this study was to measure the health-related quality of life of the Hungarian general population. The quality of life survey was part of the National Health Survey (Orszagos Lakossagi Egeszsegfelmeres, OLEF) conducted in year 2000 on a representative sample of 5503 members of the general population. RESULTS: Showed that, among the five health-related quality of life dimensions measured by the EQ-5D questionnaire, pain/discomfort had the highest (38%) prevalence. More than 60% of the population over 60 years of age suffer from pain or discomfort. In international comparison, anxiety/depression also had high prevalence in Hungary when compared to other developed countries. More than third of the Hungarian adult population today is depressed or suffer from anxiety. The analysis also highlighted the important association between socio economic factors and health-related quality of life. According to the applied model, older age, being female, lower education, and lower income are the main determinants of worse health-related quality of life. When quality of life is measured on a 0-100 scale, each additional life year is associated with a decrease of 0.4 units of quality of life, being female is associated with 3 units lower quality of life. Each 10,000 Ft increase in mean household income improves quality of life with 0.5 unit. People with higher education have 5 units higher quality of life than those below medium level education. CONCLUSION: Findings of this study provide important baseline data for future burden of illness, health economic, and health policy analyses in Hungary. PMID- 14528839 TI - [Alcohol-induced gastrointestinal diseases]. AB - Alcohol induced gastrointestinal diseases are common and significant, and may lead to early death. The annual death caused by alcoholic liver disease and pancreatitis in Hungary is up to 8000. Metabolites, mainly acetaldehyde and free radicals are responsible for the injury. Although the alcohol itself is not carcinogenic, some maligancies are more common among alcoholics. AIM AND METHODS: In this review the hepatic and extrahepatic metabolism of alcohol, the epidemiology, pathomechanism, clinical signs of the alcohol induced organ damages and the treatment options are summarized. RESULTS: Type IV alcoholdehydrogenase in the stomach has a role in the first pass metabolism. The liver is the main place of the metabolism. If the amount of alcohol exceeds the metabolising capacity, the toxic substances cause lipidperoxidation, membrane and organ injury. Alcohol metabolism is different in the pancreas, the activity of fatty acid ethanol ester synthase activity is more pronounced. In alcoholics the leukoplakia, oropharingeal carcinoma, oesophagitis, Mallory-Weiss syndrome, liver cirrhosis related oesophageal variceal bleeding, chronic gastritis, liver diseases, pancreatitis, bowel motility disorders, malabsorption and colorectal carcinoma are more frequent. The fatty liver remains reversible for a long. Following steatohepatitis, fibrosis, cirrhosis and liver carcinoma may develop. Despite growing knowledge of the role of endotoxins, cytokines, nutritional, immunological and genetic factors, it is still unknown why the given disease will develop in a patient, and there is no parameter for determining the point of irreversibility of the alterations. The possibilities for medical treatment are limited, since some patients do not cooperate properly, and on the other hand, the drugs and measurements can control only a part of the whole process (antioxidants, anti-inflammatory drugs, monoclonal anti-cytokine antibodies), and are appropriate only for the treatment of complications as ascites, portal hypertension, oesophageal varices, portosystemic encephalopathy, malabsortion, infections, vitamin deficiency. CONCLUSIONS: Therefore the prevention and abstinence are very important. The task of the general practitioner and the role of the cooperation among the patient, the physician, the family and the community are very important. PMID- 14528841 TI - [Therapeutic safety of antimycotics in the treatment of systemic fungal infections]. AB - The authors summarize the most important data of antimycotics used in the praxis. The indications of these drugs as well as the absorption, the antimycotic mechanism, the side-effects and drug-interactions of antifungal agents are discussed. The pathomechanism of amphotericin B, flucytosine, fluconasole, ketoconasole, itraconasole and the possibilities of their clinical administrations, further their side-effects and interactions are demonstrated in details. The most beneficial characteristics and more important interactions of these drugs are shown in tables for easier application in the everyday therapy. PMID- 14528842 TI - [Practical aspects of Doppler sonography in obstetrics]. AB - Doppler sonography has become a principal diagnostic method of antenatal care in Hungary during the past decade. Consequently, the perinatal outcome of high-risk pregnancies has significantly improved. Evaluation of the fetal circulation facilitates clinical diagnosis, antenatal and subnatal fetal surveillance, arrangement of functional tests and optimal timing of delivery. Advanced clinical efficacy is based on identical practical aspects and standardized routine. Based on their clinical experience and the published scientific data authors propose practical guidelines for Doppler evaluation of the uteroplacental and the fetal arterial and venous circulation. Summary of the blood flow velocity waveform analysis, and the steps of useful application offer consistent clinical comprehension and proper diagnosis. PMID- 14528843 TI - [Treatment of atrial fibrillation and flutter]. PMID- 14528844 TI - [Treatment of atopic dermatitis]. PMID- 14528845 TI - [175th anniversary of the birth of Henry Dunant]. PMID- 14528846 TI - Where is the holistic safety net for our children? PMID- 14528847 TI - Cultural constructions of health and illness. Recent cultural changes toward a holistic approach. AB - Nurses have defined their domain as holistic with a focus of promoting health. Cultural conceptualizations of health and illness have been changing toward a more complex understanding enriched by perspectives from many disciplines. Health, a more holistic concept than disease, has invited expanded cognitive constructions that accommodate an understanding of the biology, environment, social position, and the role of the mind, culture, spirituality, race, and sex. These have changed our metaphors of the body and expanded the focus to the health illness experience. These areas are discussed with implications for nursing. PMID- 14528848 TI - Restoring the patient's voice. The therapeutics of illness narratives. AB - Reflective and insightful autobiographical accounts of illness not only illuminate fundamental disruptions in selfhood and continuity of life that accompany illness, but authors of such accounts also maintain that narration is an important way to make sense of an illness episode, to restore personhood and connectedness, and to reclaim the illness experience from the medical meta narrative. That witnessing and helping to order illness narratives can be a caring/healing nursing practice modality with significant healing potential is supported both by narrative theory and by nursing's theoretical and philosophical legacy. The challenge for the nurse guided by narrative ideas is to give primacy to the patient's voice, to listen for meaning rather than for facts, and to provide a relationship enabling the evolution of the patient's story. PMID- 14528849 TI - Women aging with childhood onset disability. A holistic approach using the life course paradigm. AB - Women with childhood onset disabilities (CODs) are living longer and it is time for holistic nurses to focus on understanding such women from a life course perspective. As women with CODs live into later adulthood, nurses must be prepared to anticipate and assist as needed. Without studies designed to understand the life course of these women, nursing can neither gain a clear understanding of the problems they face with aging nor help prevent the high number of social inequities, secondary conditions, and comorbidities experienced. To begin nursing's exploration of the problems faced by women with CODs, the relevant historical changes and literature associated with aging with a COD are reviewed. Next, the life course paradigm is critically analyzed in relation to the lives of these women. In conclusion, it is posited that the life course paradigm provides the most useful guide for a holistic understanding of women with CODs. PMID- 14528850 TI - Holistic healing for women with breast cancer through a mind, body, and spirit self-empowerment program. AB - This article reports results of an integrated mind-body-spirit self-empowerment program for breast cancer survivors. Fifty-one women at various stages of breast cancer completed a series of eclectic lessons offered in a support group format. The program followed an integrated and cumulative lesson plan that progressively and systematically introduced multiple strategies for creating a balance among mental, emotional, spiritual, and physical health. The program's goals were to enable participants to experience a reduction in distress, improve perceived quality of life, reach a deeper sense of meaning and purpose in life, and experience a greater sense of perceived wellness. Self-assessments were obtained on four well-documented measures relating to both pre- and postprogram participation. Differences in pre- and postscores showed statistically significant improvement and large estimated effect sizes on all four measures. Participants' written comments provide examples of the scope and benefits of the program. PMID- 14528851 TI - Psychosocial antecedents of hostility in persons with coronary heart disease. AB - Although it is known that hostility precedes coronary heart disease (CHD), little is known about factors that influence the development and progression of hostile characteristics. The relations among hostility, self-esteem, self-concept, and psychosocial residual were conceptualized within the modeling and role-modeling theoretical framework and examined in a sample of 85 persons with CHD. There were significant associations between all variables. Regression analyses revealed that self-esteem, mistrust residual, isolation residual, and self-concept contributed significantly, accounting for 31% of the variation in hostility scores. These findings provide support for the belief that the development of hostility in persons with CHD is related to beliefs and attitudes about the self and others. Persons with self-esteem need deficits, and a subsequent build up of negative psychosocial residual, have poor self-concepts. This poor self-concept is hostile in nature and reflects a mistrust of others and a deep sense of isolation. PMID- 14528852 TI - Caring presence. Delineation of a concept for holistic nursing. AB - Interest in presence within a caring relationship has gained renewed attention in the past decade. For five decades, presence has been implicit and explicit in nursing theory. Recognizing caring presence as important to nursing, the 13th Annual Caring Research Conference in 1992 spotlighted the topic. Yet few publications appeared in nursing literature between the years 1980 to 1991. Since that time, interest in and literature about the topic has greatly increased. The elusive nature of the phenomenon, however, makes it difficult to define. This article presents a review of literature of presence and suggests a working definition that could be used as the basis for research. Placing presence within the context of caring provides a way for the holistic nurse to offer caring healing ways of being and relating. PMID- 14528853 TI - Perspectives. PMID- 14528854 TI - Intraoral acrylic splints for maxillofacial fracture repair. AB - Maxillofacial fractures in dogs and cats occur secondary to vehicular trauma, falls, kicks, gunshots, and fights with other animals. Pathologic mandibular fracture may occur secondary to periodontal disease, neoplasia, and metabolic diseases. The primary objective for repair of maxillofacial fractures in small animals is return to normal function. Therefore, it is necessary to maintain occlusal alignment while providing adequate stability for bony union. Basic principles of maxillofacial fracture repair include anatomic reduction and restoration of occlusion, application of a stable fixation to neutralize negative forces on the fracture, gentle handling of soft tissues, avoidance of iatrogenic dental trauma, extraction of diseased teeth within the fracture line, and minimizing excessive soft tissue elevation. This review article will describe the application of intraoral acrylic splints for maxillofacial fracture repair. PMID- 14528855 TI - Compound odontoma in three dogs. AB - Three young, female dogs were operated for compound odontoma. All tumors were considered stage III with treatment consisting of partial mandibulectomy or maxillectomy. Microscopic examination of the resected tissue confirmed the diagnosis. Relatively aggressive, resective surgery resulted in prolonged tumor free intervals. PMID- 14528856 TI - Dental implant replacement of the mandibular first molar tooth in a dog. AB - A new dental implant system was used to replace the mandibular right first molar tooth in an eleven-month-old male/intact, utility trained German shepherd dog. The permanent mandibular right first molar tooth had been extracted as treatment for an extensive carious lesion when the dog was 9-months of age. There were no complications associated with placement of the dental implant. However, peri implant osteomyelitis occurred secondary to a traumatic oral wound 6-months following implant placement. The 17-month post-operative examination indicated that the implant system used in this case could be maintained in a working dog that uses extreme bite forces. However, periodontal inflammation and vertical bone loss exposing the implant fixtures were noted during oral examination. Further clinical applications are required to determine if the periodontal inflammation and vertical bone loss noted in this case were complications associated with the implant, maturity of bone at the time of implant fixture placement, general biting/chewing forces placed on carnassial teeth, or the oral trauma that occurred 6-months following implant placement. PMID- 14528857 TI - Management of rostral mandibular fracture including lateral luxation of a mandibular canine tooth in a dog. PMID- 14528858 TI - The owner-animal-environment triad in the treatment of canine periodontal disease. AB - In a perfect world, all veterinarians and veterinary dental technicians would understand periodontal disease as well as the dental specialist. They would all be able to recognize the early signs of periodontal disease and recommend treatment to prevent its progression. The owners would have the financial resources, time, and desire to maintain their pet's oral health. The dogs would all be calm and compliant with home care and have no particular anesthetic risks. Unfortunately, this is not a perfect world. Some veterinarians do not understand periodontal disease any better that I understand cosmic string theory. Some owners have limited financial resources and are not particularly committed to their pet's oral health. Some animals will not tolerate any type or manner of home care. Given that animals, their mouths, and their owners come in an infinite variety of shapes and sizes (figuratively and literally), how can we talk about the treatment of periodontal disease as if it is a single condition with a single treatment, or even only a few treatment options? Each owner, animal, and its environment must be assessed on an individual basis to develop a treatment plan that is reasonable and attainable based on the unique circumstances of each case. So, what should be the goal when treating periodontal disease? Is it the preservation of all teeth at all costs? Is it the preservation of important teeth if the costs can be kept reasonable? I would suggest that the overriding goal of periodontal treatment should be the elimination and prevention of oral infection and oral pain. In the domestic environment, dogs have no real need to defend territory. They have no need to prehend and kill live prey animals. In short, the domesticated pet dog does not need teeth at all. This may seem like an odd statement for a veterinary dentist to make, but I feel quite strongly that a dog is far better off having no teeth than having bad teeth. My preference is that a dog should have a full set of healthy, functional teeth, but preserving bad teeth in the face of a poor or questionable prognosis serves no positive purpose. PMID- 14528859 TI - Intraoral acrylic splint application. PMID- 14528860 TI - The commercialization of education. Are we guilty as charged? PMID- 14528861 TI - Take time to teach thoughtfully. PMID- 14528862 TI - Communication skills for end-of-life nursing care. Teaching strategies from the ELNEC curriculum. AB - The End of Life Nursing Education Consortium (ELNEC) is a train-the-trainer educational program that helps nursing faculty integrate care of the dying patient and the patient's family into the nursing curriculum. This article presents pedagogical techniques derived from the ELNEC teaching module on communication and key content areas for developing communication skills in end-of life care. Competent end-of-life nursing care begins with an understanding of communication techniques and their potential impact on care. PMID- 14528863 TI - Incorporating concepts of end-of-life care into a psychiatric nursing course. AB - Providing quality care to patients who are at the end of life is recognized as an important aspect of health care. Nurse educators are in a unique position to influence future care by imparting essential information on this topic to students during their basic nursing education. The author offers support for the suggestion that select end-of-life concepts be included in psychiatric-mental health nursing courses and presents suggestions for incorporating such concepts into psychiatric nursing clinical learning experiences. PMID- 14528864 TI - Multidisciplinary, interdisciplinary, and transdisciplinary Educational models and nursing education. AB - In an effort to prepare health care professionals for the team-based work environment that exists in health care delivery systems today, some nursing faculty may consider collaborative, team-taught courses that integrate faculty and students from various disciplines. To assist nursing faculty in making an informed decision about integrated curriculum development and course implementation, multidisciplinary, interdisciplinary, and transdisciplinary educational teams are defined. Examples are offered that reflect these three integrated educational team models. Finally, the benefits and potential problem areas that result from team initiatives are briefly reviewed. PMID- 14528865 TI - Collaborative learning experiences for nursing students in environmental health. AB - Recognition that the environment is a determinant of health is deeply rooted in the heritage of nursing. Environmental hazards and their effects on health rarely have simple solutions that can be managed by one discipline alone. Environmental health issues are complex in nature and the science is developing rapidly. Primary prevention strategies often involve the participation of professionals from fields other than nursing, and interdisciplinary, collaborative efforts are usually required. This article describes learning experiences about local environmental health issues, several of which were designed to initiate and foster collaboration between baccalaureate nursing students and graduate students in public health. PMID- 14528866 TI - Caring as an imperative for nursing education. AB - Caring is a universal need that is an important component in the delivery of nursing care. Nurse educators face the challenge of teaching the value of caring as a necessary part of nursing. Watson's theoretical framework, which focuses on interpersonal and transpersonal processes in human care, presents an effective model in understanding the concept of caring (1). Krathwohl, Bloom, and Masia's model of affective domain provides a taxonomy of affective competencies that guides the process of value development (2). Both models enhance our understanding of caring and provide the theoretical foundation for integrating caring into nursing education. This article describes an associate degree program's effort to thread the concept of caring across its curriculum. It includes an overview of the steps used for integrating caring in individual clinical courses and emphasizes teaching/learning and assessment strategies used in the educational process. PMID- 14528867 TI - Take a chance on nursing mentorships. Enhance leadership with this win-win strategy. AB - Mentoring in nursing traditionally occurs at the master's or doctoral level, but the mutual career enhancement gained by such an alliance can be experienced at all stages of a nursing career. This article presents e-mail examples to illustrate antecedent relationship factors and the progression of phases in a mentorship between a clinically experienced RN and a doctorally prepared nursing professor. Application of this concept from a business model reveals parallels to nursing and blends perspectives from both disciplines. Also discussed are perceptual and tangible outcomes of mentorships and strategies to foster this positive approach. PMID- 14528868 TI - [Important recognition of reviewed published articles in international fields]. PMID- 14528869 TI - Healthcare needs of older prisoners are neglected. PMID- 14528870 TI - The policy position within Irish nursing. PMID- 14528871 TI - Intravenous fentanyl for cancer pain: a "fast titration" protocol for the emergency room. AB - Patients with cancer sometimes are admitted to the emergency room due to severe pain. Despite the fact that morphine's hydrophilicity can delay its peak effects after intravenous administration up to 30 minutes, it is still the most commonly used opioid during cancer pain emergencies. Fentanyl is a synthetic, lipophilic opioid, more potent than morphine, and achieves peak effects after intravenous administration in 5 minutes. According to our observations, intravenous fentanyl could be safely used in the emergency room to treat patients who need fast titration of an opioid to control their pain. In our study, fentanyl was employed in a four-step protocol to treat patients admitted to our palliative care emergency room due to severe pain, regardless of the previous use of morphine at home. Titration with intravenous fentanyl was successfully employed in 18/18 (100%) of patients, with an average time for pain control at about 11 minutes, and without relevant adverse effects. We conclude that intravenous fentanyl could be safely used for severe cancer pain when rapid titration is being considered. PMID- 14528872 TI - Bibliography. Current world literature. Lymphoma. PMID- 14528873 TI - Bibliography. Current world literature. Cancer in AIDS. PMID- 14528874 TI - [Manuscript language and agreement concerns]. PMID- 14528875 TI - Severe acute respiratory syndrome (SARS) in Hong Kong. AB - Severe acute respiratory syndrome (SARS) is a recently recognized and highly contagious pneumonic illness, caused by a novel coronavirus. While developments in diagnostic, clinical and other aspects of SARS research are well underway, there is still great difficulty for frontline clinicians as validated rapid diagnostic tests or effective treatment regimens are lacking. This article attempts to summarize some of the recent developments in this newly recognized condition from the Asia Pacific perspective. PMID- 14528876 TI - The effect of parental smoking on lung function and development during infancy. AB - While the adverse effects of parental smoking on respiratory health during childhood are well recognized, its potential impact on early lung development is less clear. This review summarizes current evidence on the effect of parental smoking on lung function during infancy. It is difficult to separate the effects of pre- and postnatal exposure, since the majority of mothers who smoke in pregnancy (currently around 30% worldwide) continue to do so thereafter. Nevertheless, measurements undertaken prior to any postnatal exposure have consistently demonstrated significant changes in tidal flow patterns in infants whose mothers smoked in pregnancy. While there is, as yet, no convincing evidence from studies in human infants that smoking during pregnancy is associated with increased airway responsiveness at birth, many studies have demonstrated a reduction in forced expiratory flows (on average by 20%) in infants exposed to parental smoking. While maternal smoking during pregnancy remains the most significant source of such exposure and is likely to be responsible for diminished airway function in early life, continuing postnatal tobacco smoke exposure will increase the risk of respiratory infections, the combination of both being responsible for the two- to fourfold increased risk of wheezing illnesses observed during the first year of life in infants whose parents smoke. These findings emphasize the need to keep infants in a smoke-free environment both before and after birth, not least because of growing awareness that airway function in later life is largely determined by that during foetal development and early infancy. PMID- 14528877 TI - History of tobacco and health. AB - Tobacco comes from plants that are native to the Americas around Peru and Ecuador,where it has been found since prehistoric times. It was brought back to Europe by early explorers where it was adopted by society and re-exported to the rest of the world as European colonization took place. Smoking tobacco in pipes of one sort or other gave way to handmade and then manufactured cigarettes, especially during the First World War. Smoking rates increased dramatically during the 20th century in developed countries until recently and rates are still increasing in underdeveloped countries. An epidemic of smoking-related diseases has followed the prevalence of smoking. Scientific knowledge of the harmful effects of active tobacco smoking has accumulated during the past 60 years since early descriptions of the increasing prevalence of lung cancer. The first epidemiological studies showing an association between smoking and lung cancer were published in 1950. In 1990 the US Surgeon General concluded that smoking was the most extensively documented cause of disease ever investigated but governments worldwide have been ambivalent and slow in taking action to reduce smoking. Tobacco smoking is now agreed to be a major cause of a vast number of diseases and other adverse effects. Since the 1980s passive smoking including exposure in utero has also been implicated as a significant cause of numerous diseases. In response, the tobacco industry has managed to forestall and prevent efforts to control this major health problem. PMID- 14528879 TI - The doc is out. Tufts-NEMC looks at nonphysicians to fill CEO slot. PMID- 14528878 TI - Bronchopulmonary afferent nerves. AB - Vagal afferent nerves are the primary communication pathways between the bronchopulmonary system and the central nervous system. Input from airway afferent nerves to the CNS is integrated in the brainstem and ultimately leads to sensations and various reflex outputs. Afferent nerves innervating the airways can be classified into various distinct phenotypes. However, there is no single classification scheme that takes all features, including conduction velocity, cell body diameter, ganglionic origin, and stimuli to which they respond (modality) into account. At present, bronchopulmonary afferent nerves are typically considered to belong to one of three general categories, namely C fibres, rapidly adapting stretch receptors (RARs), and slowly adapting stretch receptors (SARs). As our understanding of bronchopulmonary afferent nerves continues to deepen, we are likely to see more sophisticated classification schemes emerge. It is clear that the function of afferent fibres can be substantively influenced by airway inflammation and remodelling. The perturbations and perversions of afferent nerve function that occur during these states almost certainly contributes to many of the signs and symptoms of inflammatory airway disease. A more lucid characterization of bronchopulmonary afferent nerves, and a better understanding of the mechanisms by which these nerves influence pulmonary physiology during health and disease anticipates future research. PMID- 14528880 TI - Cutaneous and pulmonary sarcoidosis in a Hong Kong Chinese woman with silicone breast prostheses. AB - Sarcoidosis is rarely reported in Hong Kong. We report cutaneous and pulmonary sarcoidosis in a 54-year-old Chinese woman, who presented with papular lesions over the face and neck. She had silicone breast augmentation surgery 4 years earlier. Skin biopsy revealed granulomatous inflammation and anti-tuberculosis treatment was started empirically but stopped 2 months later owing to a poor response. A right supraclavicular lymph node was aspirated and revealed granulomatous inflammation. The CXR was normal initially but subsequently showed diffuse reticulonodular opacities and a small right-sided pleural effusion. High resolution CT of the thorax showed mediastinal lymphadenopathy and diffuse perilymphatic nodular opacities consistent with sarcoidosis. Sputum mycobacterial culture was negative. Fibreoptic bronchoscopy showed no endobronchial lesion but the transbronchial biopsy showed granulomatous inflammation with no evidence of infection, malignancy or foreign body. Pulmonary function tests were normal except for impairment of transfer factor. One year later, most of the cutaneous lesions had healed spontaneously. The CXR showed partial improvement of the right pleural opacification but little change in the lung field. The features of sarcoidosis and its association with silicone are reviewed. PMID- 14528881 TI - Lights out. Hospitals operate as usual during power outages. PMID- 14528882 TI - First European case of serotype A MATa Cryptococcus neoformans infection. PMID- 14528883 TI - Trans-fatty acid composition in diets: what should dietetics professionals know? PMID- 14528884 TI - Results of the Home Food Safety--It's in Your Hands 2002 survey: comparisons to the 1999 benchmark survey and Health People 2010 food safety behaviors objective. PMID- 14528885 TI - A case of pulmonary veno-occlusive disease associated with systemic sclerosis. AB - The case of a patient with pulmonary veno-occlusive disease associated with systemic sclerosis is reported. The patient presented with progressive dyspnoea. Echocardiography and cardiac catheterization study demonstrated right-sided heart failure. The CXR suggested pulmonary hypertension and interstitial pulmonary oedema. We suspected pulmonary veno-occlusive disease based on radiological and haemodynamic findings. Treatment with prednisolone resulted in a reduction in pulmonary arterial pressure and CXR findings improved 2 months later, but no further effect was observed. The patient died 7 months later and at autopsy the lungs showed prominent thickening of the interlobular septa and small branches of pulmonary veins showed intimal thickening. PMID- 14528886 TI - Endobronchial cryptococcosis in AIDS. AB - Although cryptococcosis is a common thoracic complication among patients with AIDS, endobronchial abnormalities have rarely been reported. A 45-year-old man presented with a productive cough, fever, and headache. His CD4+ cell count was 7/mm3 and testing for antibodies to HIV-1 was positive. Radiological examination revealed consolidation in the left lung, including cavitation. Bronchoscopic examination demonstrated white, slightly raised, plaque-like lesions in the trachea and left bronchi. Histopathological examination of endobronchial biopsy specimens revealed granulation tissue with abundant encapsulated yeast in the tissue. Cryptococcus neoformans was cultured from the bronchial lavage specimen. Cryptococcal infection should be included in the differential diagnosis of endobronchial abnormalities in patients with AIDS. PMID- 14528887 TI - A case of completely resolved pneumatocoeles in desquamative interstitial pneumonia. AB - Desquamative interstitial pneumonia (DIP), also known as alveolar macrophage pneumonia (AMP), represents a subset of idiopathic interstitial pneumonia that responds better to steroids and has a more favourable prognosis than usual interstitial pneumonia. Recently, we encountered a case of DIP with the formation of multiple pulmonary cysts during corticosteroid maintenance treatment. After the introduction of cyclophosphamide, the cysts gradually disappeared. This complete resolution is believed to have resulted from the clearance of check valve-like bronchiolar obstructions that may be another interesting terminal airway pathology in DIP. PMID- 14528888 TI - Antinuclear antibody positive pleural effusion in a patient with tuberculosis. AB - A patient with tuberculosis presented with a pleural effusion that was highly positive for antinuclear antibody (ANA). The pleural fluid autoimmune profile was positive for ANA IgG at a titre of 1 : 1280. Antibodies to double-stranded DNA were not detected in the pleural fluid or in serum. The serum autoimmune profile was positive for ANA IgG at 1 : 160 and IgM at 1 : 40. Pleural fluid was positive on culture for Mycobacterium tuberculosis after 8 weeks. Pleural biopsy for histology showed chronic inflammation and culture revealed no growth. The pleural fluid resolved with the anti-tuberculous treatment, and signs and symptoms of systemic lupus erythematosus or malignancy did not occur, which suggests that tuberculous pleural effusion is one of the causes of high ANA in pleural fluid. PMID- 14528889 TI - Management of idiopathic pulmonary haemosiderosis in pregnancy: report of two cases. AB - Idiopathic pulmonary haemosiderosis (IPH) is a rare cause of alveolar haemorrhage. Its management in pregnancy is complicated and little clinical data exist on outcomes or treatment. Two patients with IPH, one with a known diagnosis prior to pregnancy (patient A) and one presenting de novo during pregnancy (patient B) are reported. Both subjects became maximally symptomatic in the third trimester and were treated with corticosteroids. Despite steroid treatment patient A had continued symptoms and was treated with azathioprine. Both patients delivered healthy female infants at 32 weeks and 38 weeks gestation, respectively. Patient B's course was complicated by the development of fatal invasive pulmonary aspergillosis 4 weeks postpartum. With improved care, more patients with IPH survive to childbearing age. Patients become most symptomatic late in pregnancy, underscoring the need for careful third trimester monitoring. These cases demonstrate that steroids are safe and that azathioprine can be used as additional therapy. When considering treatment options it is important to weigh the benefits of these medications against the potential for maternal and foetal harm. PMID- 14528890 TI - Fatal pneumonia caused by Burkholderia cepacia 9 months after resection of aspergilloma. AB - A 69-year-old man developed an episode of severe community-acquired pneumonia 9 months after resection of aspergilloma. Although Aspergillus fumigatus was also isolated in the pleural cavity, it did not invade the remaining lung parenchyma. The patient developed progressive bilateral pneumonia leading to death from respiratory failure. Burkholderia cepacia was considered as prime pathogen, as it was repeatedly cultured from sputum and tracheal secretions, as well as the autopsy lung. B. cepacia is resistant to most antibiotics, and seldom causes pneumonia in patients without cystic fibrosis or chronic granulomatous disease. The precise reason that this apparently immunocompetent patient developed B. cepacia pneumonia remains unknown. PMID- 14528892 TI - True lumen collapse assessed by epiaortic echocardiography in aortic dissection. PMID- 14528891 TI - Assessment of stent-grafting for aortic dissection using multislice computed tomography. PMID- 14528893 TI - [Control or trust? Nothing is more important than trust]. PMID- 14528894 TI - Independence and separability of volume and mass in the size-weight illusion. AB - Numerous size-weight illusion models were classified in the present article according to general recognition theory (Ashby & Townsend, 1986), wherein the illusion results from a lack of perceptual separability, perceptual independence, decisional separability, or a combination of the three. These options were tested in two experiments in which a feature-complete factorial design and multidimensional signal detection analysis were used (Kadlec & Townsend, 1992a, 1992b). With haptic touch alone, the illusion was associated with a lack of perceptual and decisional separability. When the participant viewed the stimulus in his or her hand, the illusion was associated only with a lack of decisional separability. Visual input appeared to improve the discrimination of mass, leaving only the response bias due to expectation. PMID- 14528895 TI - An assessment of response bias for the same-different task: implications for the single-interval task. AB - Two auditory amplitude discrimination experiments were conducted using the same different experimental design. Observer bias was manipulated, in the first experiment, by varying payoff matrices and, in the second, by varying prior probabilities of signal presentation. Five levels of bias manipulation and four levels of difficulty were employed in each experiment. Each observer received all combinations of bias manipulation and difficulty, but with only one of these combinations within each block of trials. Nine indices of bias were assessed by simultaneously fitting isosensitivity and isobias functions to the data and by fitting isobias functions only. Although none of the indices tested provided an exceptionally good fit to the data, two indices stood out from the rest. These were c*sd and ci, indices with isobias contours similar in shape to those for the c index derived from the yes-no task. PMID- 14528896 TI - Spatial distortions induced by multiple visual landmarks: how local distortions combine to produce complex distortion patterns. AB - Visual landmarks introduce systematic distortions into spatial short-term memory for single target positions, the exact form of the distortion depending on the spatial layout of the landmarks. In two experiments, we investigated how the combined effect of two landmarks can be predicted from the effects of individual landmarks. Participants used a mouse cursor to reproduce the positions of briefly presented targets in the context of one, the other, or both landmarks. We found that distortions near a landmark are independent of whether another landmark is present, so that remembered space is partitioned into regions dominated by single landmarks. Interestingly, the display midpoint behaves like a "virtual landmark," with its own pattern of distortion. Results are inconsistent with current models of spatial memory distortions but suggest that attentional processes can lead to enhanced fidelity of salient regions in topographical neural networks while also introducing some spatial biases. PMID- 14528897 TI - Compound self-motion perception induced by two kinds of optical motion. AB - Two kinds of flow patterns consisting of random dots were presented simultaneously to subjects to investigate whether or not two kinds of vection occur simultaneously. One pattern induces vertical linear self-translation, whereas the other induces self-rotation around a vertical axis (when either pattern is presented alone). Three sets of conditions were tested. The first condition was one in which random dots moved in a summed direction of both flow vectors. In the second condition, both flow patterns were simply overlaid, whereas in the third condition, the two kinds of flow patterns were overlaid with a depth separation produced by binocular disparity. The subjects perceived both kinds of vection simultaneously in directions opposite to those of the corresponding flow components under the first condition, whereas either vection occurred mainly under the second condition. Under the third condition, both of the flows induced each kind of vection simultaneously, despite there being no physical vector summation of dot motion. The background flow induced vection in a direction opposite to the flow direction, whereas the foreground flow induced vection in the same direction as the flow direction. These results show that induced self-translation and induced self-rotation can occur simultaneously in two ways. PMID- 14528898 TI - Attentional focus, processing load, and Stroop interference. AB - Although the effects of attentional focus and perceptual load on selective attention when targets and distractors are distinct objects that occupy separate locations are well known, there has been little examination of their role when both relevant and irrelevant information pertains to the same object. In four experiments, participants were shown Stroop color words or strings of letters in a task of speeded color identification. When the participants' attentional focus was manipulated via cue validity or precue size, greater Stroop interference was observed when the attentional focus was narrow than when it was broad. However, when the participants were induced to adopt a comparable attentional focus in a dual-task paradigm, the differential Stroop interference was eliminated. Furthermore, contrary to the prediction of the perceptual load hypothesis, different levels of processing load did not lead to differential Stroop interference. These results emphasize the importance of stimulus structure in understanding distractor processing. They indicate that when relevant and irrelevant information pertains to the same object, narrowing attentional focus increases distractor processing, and perceptual load has a negligible effect on the extent of distractor processing. PMID- 14528899 TI - Crossmodal links between audition and touch in covert endogenous spatial attention. AB - We report three experiments designed to investigate the nature of any crossmodal links between audition and touch in sustained endogenous covert spatial attention, using the orthogonal spatial cuing paradigm. Participants discriminated the elevation (up vs. down) of auditory and tactile targets presented to either the left or the right of fixation. In Experiment 1, targets were expected on a particular side in just one modality; the results demonstrated that the participants could spatially shift their attention independently in both audition and touch. Experiment 2 demonstrated that when the participants were informed that targets were more likely to be on one side for both modalities, elevation judgments were faster on that side in both audition and touch. The participants were also able to "split" their auditory and tactile attention, albeit at some cost, when targets in the two modalities were expected on opposite sides. Similar results were also reported in Experiment 3 when participants adopted a crossed-hands posture, thus revealing that crossmodal links in audiotactile attention operate on a representation of space that is updated following posture change. These results are discussed in relation to previous findings regarding crossmodal links in audiovisual and visuotactile covert spatial attentional orienting. PMID- 14528900 TI - Sensory mediation of stimulus-driven attentional capture in multiple-cue displays. AB - Three location-cuing experiments were conducted in order to examine the stimulus driven control of attentional capture in multiple-cue displays. These displays consisted of one to four simultaneously presented direct location cues. The results indicated that direct location cuing can produce cue effects that are mediated, in part, by nonattentional processing that occurs simultaneously at multiple locations. When single cues were presented in isolation, however, the resulting cue effect appeared to be due to a combination of sensory processing and attentional capture by the cue. This suggests that the faster responses produced by direct cues may be associated with two different components: an attention-related component that can be modulated by goal-driven factors and a nonattentional component that occurs in parallel at multiple direct-cue locations and is minimally affected by goal-driven factors. PMID- 14528901 TI - Goal-driven modulation of stimulus-driven attentional capture in multiple-cue displays. AB - Six location-cuing experiments were conducted to examine the goal-driven control of attentional capture in multiple-cue displays. In most of the experiments, the cue display consisted of the simultaneous presentation of a red direct cue that was highly predictive of the target location (the unique cue) and three gray direct cues (the standard cues) that were not predictive of the location. The results indicated that although target responses were faster at all cued locations relative to uncued locations, they were significantly faster at the unique-cue location than at the standard-cue locations. Other results suggest that the faster responses produced by direct cues may be associated with two different components: an attention-related component that can be modulated by goal-driven factors and a nonattentional component that occurs in parallel at multiple direct-cue locations and is minimally affected by the same goal-driven factors. PMID- 14528902 TI - Coping with phonological assimilation in speech perception: evidence for early compensation. AB - The pronunciation of the same word may vary considerably as a consequence of its context. The Dutch word tuin (English, garden) may be pronounced tuim if followed by bank (English, bench), but not if followed by stoel (English, chair). In a series of four experiments, we examined how Dutch listeners cope with this context sensitivity in their native language. A first word identification experiment showed that the perception of a word-final nasal depends on the subsequent context. Viable assimilations, but not unviable assimilations, were often confused perceptually with canonical word forms in a word identification task. Two control experiments ruled out the possibility that this effect was caused by perceptual masking or was influenced by lexical top-down effects. A passive-listening study in which electrophysiological measurements were used showed that only unviable, but not viable, phonological changes elicited a significant mismatch negativity. The results indicate that phonological assimilations are dealt with by an early prelexical mechanism. PMID- 14528903 TI - The role of spatial working memory in inhibition of return: evidence from divided attention tasks. AB - Inhibition of return (IOR) refers to a bias against returning attention to a location that has been recently attended. In the present experiments, we examined the role of working memory in IOR by introducing secondary tasks (in the temporal interval between the cue and the target) that involved a working memory component. When the secondary task was nonspatial in nature (monitoring odd digits or adding digits), IOR was present, although overall reaction times were greater in the presence of the secondary task. When the task involved a spatial working memory load (remembering the directionality of arrows or the orientation of objects), IOR was eliminated. However, when the participants had incentive to process the directionality of an arrow but did not have to use any memory system, IOR persisted at peripheral locations. Overall, the results suggest that IOR is partially mediated by a spatial working memory system. PMID- 14528904 TI - What is "marked" in visual marking? Evidence for effects of configuration in preview search. AB - Visual search for a conjunction target is facilitated when distractor sets are segmented over time: the preview benefit Watson and Humphreys (1997) suggested that this benefit involved inhibition of old items (visual marking, VM). We investigated whether the preview benefit is sensitive to the configuration of the old distractors. Old distractors changed their location prior to the occurrence of the new items, while also either changing or maintaining their configuration. Configuration changes disrupted search. The results are consistent with object based VM, which is sensitive to the configuration of old stimuli. PMID- 14528905 TI - Clinical mass spectrometry in neuroscience. Proteomics and peptidomics. AB - In this review we discuss the merits and drawbacks with the use of proteomic and peptidomic strategies for identification of proteins and peptides in their multidimensional interactions in complex biological processes. The progress in proteomics and peptidomics during the last years offer us new challenges to study changes in the protein and peptide synthesis. These strategies also offer new tools to follow post-translational modifications and other disturbed chemical processes that may be indicative of pathophysiological alteration(s). Furthermore these techniques can contribute to improvements in the diagnosis and therapy of neurodegenerative diseases, such as Alzheimer's disease, and psychiatric diseases, as depression and post traumatic stress disorders. We also consider different practical aspects of the applications of mass spectrometry in clinical neuroscience, illustrated by example from our laboratories. The new proteomic and peptidomic strategies will further enable the progress for clinical neuroscience research. PMID- 14528906 TI - The human pituitary proteome: the characterization of differentially expressed proteins in an adenoma compared to a control. AB - In order to clarify the basic molecular mechanisms that participate in the formation of human pituitary macroadenomas, this study, for the first time, describes the comparative proteomics between a pituitary adenoma tissue and a control tissue. A vertical, two-dimensional polyacrylamide gel electrophoresis system and PDQuest image analysis software were used to provide a high level of between-gel reproducibility and electrophoretic separation to accurately locate each differentially expressed protein. Mass spectrometry (MALDI-TOF and LC-ESI-Q IT) and protein databases were used to characterize each differentially expressed protein. A total of 137 differential gel spots (37 increased spot volumes, 39 decreased, 19 new and 42 lost) were found when we compared an adenoma proteome to a control proteome. Seventy-one spots (20 increased, 27 decreased, 13 new, 11 lost), representing 39 differentially regulated proteins, were identified. Five differentially regulated proteins (prolactin, cellular retinoic acid-binding protein II, G-protein beta subunit 3, secretagogin and calreticulin) were also validated with results from a comparative transcriptomics study of pituitary adenomas and controls. The functional characteristics of these differentially expressed proteins provide a differential proteomic profile between a pituitary adenoma and a control. PMID- 14528907 TI - Quantitative peptidomics of pituitary glands from mice deficient in copper transport. AB - We previously described a method of quantitating levels of peptides in Cpe(fat)/Cpe(fat) mice using affinity chromatography to isolate peptide processing intermediates and differential isotopic labeling/mass spectrometry. In the present study, we compared two different isotopic labels, acetic anhydride and succinic anhydride for detection and quantitation of peptides in wild type mice. As previously found for acetic anhydride, succinic anhydride efficiently labels all primary amines in various peptides. Of these two reagents, succinic anhydride provides better resolution between the heavy and light peaks of the labelled peptides due to a greater mass difference between the deuterated (heavy) and non-deuterated (light) form of this label (4 Da for succinate, 3 Da for acetate). Using succinic anhydride labeling, the accuracy of measuring 1:1 and 1:2 ratios of peptides in pituitary extracts was within 5% of the theoretical value for most peptides. The accuracy with succinic anhydride is comparable to the accuracy of acetic anhydride and more peptides could be detected and quantitated with succinic anhydride. The two labels were then used to examine pituitary peptides in mice with a defect in copper transport (Atp7a mice) vs wild type mice. Using succinic anhydride, 13 peptides could be detected, 12 of which matched the theoretical mass of known pituitary peptides. Five of the six peptides which contain C-terminal amide groups were significantly decreased in the Atp7a mice relative to wild type mice, whereas only one non-amidated peptide was significantly decreased in Atp7a mice. With acetic anhydride, only five peptides could be quantitated. The three peptides which contain C-terminal amide groups were decreased approximately 30% in the Atp7a mice. The selective decrease in amidated peptides in Atp7a mice is consistent with the copper-requirement of the enzyme that forms C-terminal amides. PMID- 14528908 TI - Analysis of protein induction in the CNS of SJL mice with experimental allergic encephalomyelitis by proteomic screening and immunohistochemistry. AB - Experimental allergic encephalomyelitis (EAE) is an autoimmune disease characterized by demyelination and inflammatory infiltrates in the CNS, and it is an animal model of multiple sclerosis. Piperonyl butoxide (PBO) suppresses disease in EAE mice, and it exhibits a dual effect on cytochrome P450s that manifests in a transient inhibitory phase followed by induction. In order to identify the expression of proteins associated with EAE, a proteomic screening was performed on hindbrain microsomes from control + vehicle, control + PBO, EAE + vehicle, and EAE + PBO female mice. Glucose regulated protein 94 (Grp94) and coagulation factor VIII were among the proteins identified in EAE + vehicle and EAE + PBO mice. Immunohistochemical staining of Grp94 was present in some neurons and oligodendrocytes in hindbrain sections from control animals, and in some cells within inflammatory infiltrates in EAE animals. Since Grp94 (also known as Gp96) can partake in antigen presentation and induction of proinflammatory cytokine expression, its presence in these cells suggests that it may play a role in the pathogenesis of EAE. Coagulation factor VIII is carried and protected by von Willebrand factor. Immunohistochemical staining of von Willebrand factor revealed its presence in some vessels within hindbrain sections from control animals. In EAE animals, the number of labeled vessels was significantly increased, and extracellular granular deposits were observed around labeled vessels indicating that the breakdown of the blood-brain barrier that occurs in EAE permitted its extravasation into the CNS. Additional proteins were identified in the different groups of mice by proteomic screening, but confirmation of their expression profile awaits investigations by independent measures. PMID- 14528911 TI - Proteomic analysis of oxidatively modified proteins in Alzheimer's disease brain: insights into neurodegeneration. AB - Identification of oxidized proteins in Alzheimer's disease (AD) brain is hypothesized to lead to new insights into mechanisms of neurodegeneration and synapse loss in this dementing disorder that are associated with oxidative stress. Previous studies had shown increased oxidation of proteins in AD brain, but identifying those particular proteins that were specifically oxidized using standard immunochemical methods is a daunting task when one considers how many proteins there are in brain. To address this issue, proteomics has been used to identify specifically modified proteins in AD brain. This review outlines the nature of proteomics, the proteins identified in AD brain that are specifically oxidatively modified, and provides rational consequences related to neurodegeneration and synapse loss as sequelae to loss of function, due to oxidation and consistent with the known pathological and biochemical alteration in AD brain. The use of proteomics to learn about disease mechanisms is still embryonic, but the emerging techniques of proteomics represent a promising means to elucidate mechanisms of disease at the protein level. There are limitations to proteomics, and these, too, are discussed. PMID- 14528910 TI - Deranged hypothetical proteins Rik protein, Nit protein 2 and mitochondrial inner membrane protein, Mitofilin, in fetal Down syndrome brain. AB - Down syndrome (DS) is the most common genetic disorder with mental retardation and a host of deranged proteins has already been described. Protein hunting leads to rapid accumulation of aberrant proteins and proteomics methods not only allow unambiguous identification of proteins, they are also a powerful tools to identify new or predicted proteins. We applied two-dimensional gel electrophoresis with in-gel digestion of proteins and subsequent MALDI-TOF mass spectrometrical identification and quantification of spots using specific software on cortical brain samples from 7 controls and 7 samples from fetal DS at the early second trimester. Nine hypothetical proteins were identified: three of them (4833418L03Rik protein Q9D614, mitochondrial inner membrane protein Q16891 and Nit protein 2 Q8WUF0) were significantly and about doublefold reduced in fetal DS brain. Hypothetical proteins CGI 99, FLJ10463, 70 kDa WD-repeat tumor rejection antigen homolog, KSRP, Hypothetical protein 49.6 kDa and Elongin A were comparable between groups. Domain analysis of deranged structures revealed a t_SNARE domain for the Rik protein, indicating involvement of this protein in the exocytotic-synaptic machinery impaired in DS, a CN hydrolase domain for Nit protein 2, possibly reflecting aberrant nitrilase-related metabolism and handling and an inner mitochondrial protein, extending knowledge on the mitochondrial deficit in in fetal DS early in life. PMID- 14528912 TI - CU(II)-catalyzed oxidation of Alzheimer's disease beta-amyloid peptide and related sequences: remarkably different selectivities of neurotoxic betaAP1-40 and non-toxic betaAP40-1. AB - We investigated the CuII-catalyzed oxidation of beta-amyloid peptides betaAP10-20 and betaAP40-1 by tandem mass spectrometry and compared oxidation yields and selectivities to those for betaAP1-16, betaAP1-28 and betaAP1-40, which were obtained earlier (26). While betaAP1-16, betaAP1-28 and betaAP1-40 showed an almost exclusive oxidation of His residues to 2-oxo-histidine, the selectivity pattern is changed for betaAP10-20,which shows oxidation of His but also hydroxylation of Tyr and Phe. In contrast to betaAP1-40, the reverse sequence betaAP40-1 shows a strong selectivity for the hydroxylation of Tyr31 while only negligible His oxidation is observed at early time points. These selectivity patterns show the importance of the geometry of the metal-binding site for peptide/protein oxidation. The significantly different characteristic of betaAP1 40 and betaAP40-1 with regard to metal catalyzed processes may be related to the differences in the neurotoxic properties of these sequences. PMID- 14528913 TI - Metal binding to brain-specific metallothionein-3 studied by electrospray ionization mass spectrometry. AB - Metallothionein-3 (MT-3) is a brain-specific isoform of metallothioneins, which is down-regulated in Alzheimer's disease (AD), inhibits the growth of neurons in vitro, and differs from common MTs also in gene regulation. To elucidate the differences in structure and function between MT-3 and common MTs, Zn2+ and Cd2+ binding to MT-3 and MT-1 were studied using electrospray ionization time of flight mass spectrometry (ESI TOF MS) at pH values between 7.5 and 2.7. The metal binding properties of MT-3 differ considerably from those of MT-1. After reconstitution with a metal excess, metallated MT-3 exists as a mixture of Zn7MT 3 (or Cd7MT-3, respectively) and several metalloforms with stoichiometries below and above seven. In contrast, MT-1 exists as a single Zn7MT-1 (or Cd7MT-1). Lowering of pH leads to a stepwise release of metals from metallated MT-3, first from extra sites, then from the 3-metal cluster and finally from the 4-metal cluster. At acidic pH values the 4-metal cluster of MT-3 is slightly more stable than that of MT-1. The results demonstrate higher structural plasticity, dynamics and metal binding capacity of MT-3 than of MT-1, which makes MT-3 suitable as a zinc buffer-transfer molecule in zinc-enriched neurons functioning at conditions of fluctuating zinc concentrations. PMID- 14528909 TI - Decreased brain levels of Lupus La protein and increased U5 small ribonucleoprotein-specific 40 kDa protein in fetal Down syndrome. AB - RNA-binding proteins have important role in the post-transcriptional regulation of gene expression. They are involved in events such as mRNA processing, transport, stability and translation. Studies in different species indicate that mutants with defect in RNA-binding proteins are defective in cell growth and differentiation. Expression of various RNA-binding proteins in prenatal life was analyzed by the highly sensitive two-dimensional electrophoresis coupled to matrix-assisted laser desorption ionization mass spectroscopy. No apparent change was obtained in levels of heterogeneous nuclear ribonucleoproteins (A3, C1-C2, L and M), nucleolin, polyadenylate binding protein-1, nuclear factor associated with double stranded RNA-2 and RNA-binding motif protein-4 between control and Down syndrome fetuses. By contrast, U5 small nuclear ribonucleoprotein-specific 40 kDa protein (p < 0.05) and Lupus La protein (p < 0.01) were significantly elevated and reduced, respectively in fetal DS. As a conclusion we can say U5 small nuclear ribonucleoprotein-specific 40 kDa protein appears to play important role in spliceosome assembly and disassembly, whereas La protein is involved in small nuclear riboncleoprotein complex biogenesis and transfer RNA maturation. Aberrant expression of these proteins points to the fact that dysregulation of the splicing and translation processes is apparent early in prenatal life, and may contribute to the defective growth and differentiation in Down syndrome. PMID- 14528914 TI - Mass spectrometry in the study of lysosomal storage disorders. AB - Lysosomal storage disorders represent a group of over 45 distinct genetic diseases, each one resulting from a deficiency of a particular lysosomal protein or, in a few cases, from non-lysosomal proteins that are involved in lysosomal biogenesis. A common biochemical feature of this group of disorders is the accumulation within lysosomes of undegraded or partially degraded substrates that are normally degraded within, and transported out of the lysosome. The particular substrates stored and the site(s) of storage vary with disease type and enzyme/protein deficiency. The nature of the substrate can be used to group the disorders into broad categories including the mucopolysaccharidoses, lipidoses, glycogenoses and oligosaccharidoses. These categories show many clinical similarities within groups as well as significant similarities between groups. For most lysosomal storage disorders the relationship between the stored substrates (type, amount and location) and the disease pathology is not well understood. The use of mass spectrometry and in particular tandem mass spectrometry provides a powerful tool for the investigation of stored substrates in this group of disorders. In this review we will describe the use of mass spectrometry for the analysis of stored substrates. We will discuss progress in the field, limitations of current methods, and summarise issues relating to the diagnosis and treatment of some of the more prevalent lysosomal storage disorders. PMID- 14528915 TI - Sphingolipid profile in the CNS of the twitcher (globoid cell leukodystrophy) mouse: a lipidomics approach. AB - Globoid cell leukodystrophy (Krabbe disease) is caused by mutations in galactosylceramidase, a lysosomal enzyme that acts to digest galactosylceramide, a glycolipid concentrated in myelin, and psychosine (galactosylsphingosine). Globoid cell leukodystrophy has been identified in many species including humans and twitcher mice. Several studies on human tissue have examined the lipid profile in this disease by gas, liquid or thin layer chromatography. Electrospray ionization tandem mass spectrometry combined with reverse phase HPLC has become a powerful alternative strategy, used here to compare the sphingolipid profile of pons/medulla tissue from twitcher mice with control tissue. In this lipidomics LC MS approach, we scanned for precursors of m/z 264 to obtain a semi-quantitative profile of ceramides and galactosylceramides. Sphingosine-1-phosphate, C18:0 ceramide, C22:0 ceramide and C24:0 ceramide levels were reduced in the pons/medulla of twitcher mice compared to levels in control mice at 31 and 35-37 days of age. The levels of C22:0 and C24:0 galactosylceramide were similar between twitcher and control specimens and there was a trend toward reduced levels of C24:1 galactosylceramide and C24:1 hydroxy-galactosylceramide in twitcher specimens. Psychosine, C 16:0 ceramide and C 18:0 galactosylceramide levels were increased in the CNS of twitcher mice compared to levels in control mice. These data indicate that there is a trend toward decreased levels of long chain fatty acids and increased levels of shorter chain fatty acids in galactosylceramides and ceramides from twitcher mice compared with control mice, and such changes may be due to demyelination characteristic of acute pathology. PMID- 14528916 TI - Metabolomic profiling of sphingolipids in human glioma cell lines by liquid chromatography tandem mass spectrometry. AB - Sphingolipids participate in membrane structure and signaling in neuronal cells, and an emerging strategy for control of gliomas is to inhibit growth and/or induce apoptosis using ceramide and ceramide analogs. Nonetheless, some sphingolipids (ceramides and sphingosine) induce and others (sphingosine 1 phosphate) inhibit apoptosis; therefore, when testing putative anti-cancer agents, it is critical to obtain precise knowledge of the types and quantities of not only the test compounds, but also their effects on endogenous species. Combination of liquid chromatography and tandem mass spectrometry affords a "metabolomic" profile of all of the intermediates of ceramide biosynthesis (3 ketosphinganine, sphinganine and dihydroceramides) and the direct products of ceramide metabolism (sphingomyelins and monohexosylceramides as well as sphingosine and sphingosine 1-phosphate). This method has been applied to four human glioma cell lines (LN18, LN229, LN319 and T98G), and differences in the amounts and types of sphingolipids were found. For example, LN229 and LN319 have approximately twice the sphingosine 1-phosphate of LN18 and T98G; LN229 and LN319 have more monohexosylceramides than lactosylceramides, whereas the opposite is the case for LN18 and T98G; and the fatty acyl chain distributions of the sphingolipids differ among the cell lines. The ability to obtain this type of "metabolomic" profile allows studies of how anti-cancer agents (especially sphingolipids and sphingolipid analogs) affect the amounts of these bioactive species, and may lead to a better understanding of the abnormal phenotypes of gliomas. PMID- 14528917 TI - Characterization of a recombinant molecule covalently indistinguishable from human cerebroside-sulfate activator protein (CSAct or Saposin B). AB - Humans deficient in the cerebroside-sulfate activator protein (CSAct or Saposin B) are unable to catabolize sulfatide and other glycosphingolipids leading to their accumulation and neurodegenerative disease. Clinically this usually manifests as a form of metachromatic leukodystrophy (MLD). CSAct is a small water soluble glycoprotein that apparently functions in the lysosome to solubilize sulfatide and other lipids enabling their interaction with soluble lysosomal hydrolases. CSAct activity can be measured in vitro by assay of its ability to activate sulfatide-sulfate hydrolysis by arylsulfatase A or ex vivo by its ability to functionally complement CSAct deficient fibroblast cell lines derived from MLD patients. A recombinant form of CSAct has been expressed in E. coli and processed in vitro to a form covalently indistinguishable from deglycosylated human CSAct isolated from human urine. Size-exclusion chromatography in combination with multi-angle laser-light scattering (SEC-MALLS) measurements demonstrate that both native and recombinant forms of the molecule behave as a dimer in the pH range 7.0-4.5. The CSAct activity assay showed that both recombinant and deglycosylated human urine CSAct efficiently activated sulfatide sulfate hydrolysis and provided functional complementation of CSAct-deficient cells. However, a D21N mutant form of recombinant CSAct could not functionally complement these cells despite full activity in the in vitro assay. It is concluded that while glycosylation is unnecessary for in vitro and ex vivo activity of CSAct, modification of the native N21 is necessary to prevent loss of ex vivo activity, possibly via protection from degradation. PMID- 14528918 TI - Specificity and potential mechanism of sulfatide deficiency in Alzheimer's disease: an electrospray ionization mass spectrometric study. AB - Recently, we have demonstrated that sulfatide content was substantially depleted in post-mortem brain samples from subjects with very mild Alzheimer's disease (AD) relative to age-matched controls. However, it is unknown if the observed sulfatide deficiency is AD-specific and what mechanism(s) lead to this depletion. By exploiting the advantages of electrospray ionization mass spectrometry techniques, we examined the specificity and a potential mechanism of sulfatide deficiency in AD in the study. In contrast to the sulfatide depletion observed in AD, it was found that the sulfatide content in post-mortem brain samples from subjects with Parkinson's disease and dementia with Lewy bodies was either higher than or comparable to that observed from controls, respectively, suggesting that sulfatide deficiency is likely specific to AD. Examination of lipid alterations in cultured embryonic rat brain oligodendrocytes treated with amyloid-beta peptide demonstrated that there was no alteration in sulfatide content up to a 24 hr interval after amyloid-beta addition/treatment. However, there were significant decreases in plasmenylethanolamine and increases in sphingomyelin content in the same study. These findings suggest that sulfatide deficiency in AD is unlikely mediated directly by amyloid-beta peptide accumulation. Thus, these results illustrate the specificity of sulfatide deficiency in AD and exclude amyloid-beta accumulation as a factor directly contributing to sulfatide deficiency in AD. PMID- 14528919 TI - 13C enrichment of extracellular neurotransmitter glutamate in rat brain--combined mass spectrometry and NMR studies of neurotransmitter turnover and uptake into glia in vivo. AB - 13C-enrichment analysis of glutamate in the extracellular fluid (GLU(ECF): 2-3 microM) by gas-chromatography/mass-spectrometry (GCMS) was combined with in vivo NMR observation of whole-brain GLU (approximately10 mM) to study neurotransmitter uptake. Brain GLU C5 was 13C-enriched by intravenous [2,5-13C]glucose infusion. GLU(ECF) was collected by microdialysis from the cortico-striatal region of awake rats. The 13C-enrichment of basal dialysate GLU C5 during 0.75-1.25 hr of infusion was 0.263 +/- 0.01, very close to the enrichment of whole-brain GLU C5. The result strongly suggests that dialysate GLU consists predominantly of neurotransmitter GLU. For selective 13C-enrichment of neurotransmitter GLU, the whole-brain 13C-enrichment was followed by [12C]glucose infusion to chase 13C from the small glial GLU pool. This leaves [5-13C]GLU mainly in the large neuronal metabolic pool and the vesicular neurotransmitter pool. The uptake of synaptic [5-13C]GLU(ECF) into glia and metabolism to glutamine (GLN) were monitored in vivo by NMR observation of [5-13C,15N]GLN formed during 15NH4Ac infusion. The rate of GLN synthesis, derived from neurotransmitter GLU(ECF) (which provided 80-90% of the substrate) was 6.4 +/- 0.44 micromol/g/hr. Hence, the observed rate represents a reasonable estimate for the rate of glial uptake of GLU(ECF), a process that is crucial for protecting the brain from GLU excitotoxicity. PMID- 14528920 TI - Simultaneous gas chromatographic-mass spectrometric determination of dopamine, norsalsolinol and salsolinol enantiomers in brain samples of a large human collective. AB - Using a solid-phase extraction procedure, an enantioselective derivatization and a gas chromatographic-mass spectrometric method, the levels of dopamine (DA) and of the dopamine-derived tetrahydroisoquinoline alkaloids (R)/(S)-salsolinol (SAL) and norsalsolinol (NorSAL) were determined in human brain samples. A complex pre analytical synthesis of reference substances as well as deuterated internal standards allowed the standardized and reproducible analysis. In this study, to our best knowledge for the first time, the regional distribution of (R)-SAL and (S)-SAL, as well as NorSAL is examined systematically in a large collective of human brain samples obtained by autopsy. The material comprises 91 brains and 8 standardized specimens in each case. Anatomical concentration differences and no ubiquitous occurence were encountered. Significant amounts of (R)-SAL, (S)-SAL and NorSAL were only found in dopamine-rich areas of the basal ganglia, whereas in other regions of the brain no tetrahydroisoquinolines were detected. These findings suggest that the concentration of the substrate dopamine may determine the alkaloid level during in vivo formation. In our opinion, non-enzymatic formation of SAL via the Pictet-Spengler reaction reveals both the SAL enantiomers. An additional enzymatic synthesis of only (R)-SAL could explain the predominant occurrence of this enantiomer. Especially in the nucleus caudatus, the concentrations of DA, SAL and NorSAL decreased significantly with rising age, which may be consistent with apoptotic effects of ageing. Our data can serve as reference for other studies in humans concerning the etiology of alcoholism or other neurodegenerative diseases with the involvement of tetrahydroisoquinolines. PMID- 14528922 TI - Irradiation stability of silicon photodiodes for extreme-ultraviolet radiation. AB - Photodiodes are used as easy-to-operate detectors in the extreme-ultraviolet spectral range. At the Physikalisch-Technische Bundesanstalt photodiodes are calibrated with an uncertainty of spectral responsivity of 0.3% or less. Stable photodiodes are a prerequisite for the dissemination of these high-accuracy calibrations to customers. Silicon photodiodes with different top layers were exposed to intense extreme-ultraviolet irradiation. Diodes coated with diamondlike carbon or TiSiN proved to be stable within a few percent up to a radiant exposure of 100 kJ/cm2. The changes in responsivity could be explained as being due to carbon contamination and to changes in the internal charge collection efficiency. In ultrahigh vacuum, no indication of oxidation was found. PMID- 14528923 TI - Three-dimensional image construction by curved surface scratches. AB - Scratches on the surface of transparent or nontransparent media reflect, refract, or diffract incident light. Under parallel illumination each length element of a scratch produces a fan beam (in the absence of diffuse scattering). Looking at a curved scratch, the right and left eyes are hit by different fan beams. Thus each eye sees a separate light spot on the scratch, which is the origin of the fan beam. In certain circumstances these spots can be stereoscopically combined by both eyes and only one light spot in space is seen. Three-dimensional images can be created by a large number of such spots originating from circular or semicircular scratches. These scratches can easily be produced on the surface of Plexiglas or other materials by using a compass. Some experiments and the theory of the so-called scratchograms are given. A comparison with holographic images is made. PMID- 14528921 TI - Immunocytochemical and molecular data guide peptide identification by mass spectrometry: orcokinin and orcomyotropin-related peptides in the stomatogastric nervous system of several crustacean species. AB - In order to identify new orcokinin and orcomyotropin-related peptides in crustaceans, molecular and immunocytochemical data were combined with matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). In the crayfish Procambarus clarkii, four orcokinins and an orcomyotropin related peptide are present on the precursor. Because these peptides are highly conserved, we assumed that other species have an identical number of peptides. To identify the peptides, immunocytochemistry was used to localize the regions of the stomatogastric nervous system in which orcokinins are predominantly present. One of the regions predominantly containing orcokinins was a previously undescribed olive-shaped neuropil region within the commissural ganglia of the lobsters Homarus americanus and Homarus gammarus. MALDI-TOF MS on these regions identified peptide masses that always occur together with the known orcokinins. Seven peptide ions occurred together in the peptide massspectra of the lobsters. Mass spectrometric fragmentation by MALDI-MS post-source decay (PSD) and electrospray ionization quadrupole time-of-flight mass spectrometry (ESI Q-TOF MS) collision-induced dissociation (CID) were used in the identification of six of these masses, either as orcokinins or as orcomyotropin-related peptides and revealed three hitherto unknown peptide variants, two of which are [His13] orcokinin ([M+H]+ = 1540.8 Da) and an orcomyotropin-related peptide FDAFTTGFGHN ([M+H]+ = 1213.5 Da). The mass of the third previously unknown orcokinin variant corresponded to that of an identified orcokinin, but PSD fragmentation did not support the suggested amino acid sequence. CID analysis allowed partial de novo sequencing of this peptide. In the crab Cancer pagurus, five orcokinins and an orcomyotropin-related peptide were unambigously identified, including the previously unknown peptide variant [Ser9-Val13]-orcokinin ([M+H]+ = 1532.8 Da). PMID- 14528924 TI - Sensitivity errors in interferometric deformation metrology. AB - Interferometric measurement techniques such as holographic interferometry and electronic speckle-pattern interferometry are valuable for measuring the deformation of objects. Conventional theoretical models of deformation measurement assume collimated illumination and telecentric imaging, which are usually only practical for small objects. Large objects often require divergent illumination, for which the models are valid only when the object is planar, and then only in the paraxial region. We present an analysis and discussion of the three-dimensional systematic sensitivity errors for both in-plane and out-of plane interferometer configurations, where it is shown that the errors can be significant. A dimensionless approach is adopted to make the analysis generic and hence scalable to a system of any size. PMID- 14528925 TI - Quasi-achromatic laser Doppler anemometry systems based on a diffractive beam splitter. AB - We propose a new beam-splitter system that makes it possible to use nonstabilized laser diodes for laser Doppler anemometry (LDA) systems by making the system wavelength independent. The beam splitter consists of two linear diffraction gratings that produce two parallel beams with a beam spacing that is wavelength dependent. This ensures passive wavelength compensation for the fringe spacing in the measurement volume. One can choose the distance between the two parallel beams by changing the distance between the two gratings, whereas the distance to the measurement volume can be designed by choice of a condensing lens with the proper focal length. This means that the system can be designed to have a desired fringe spacing in the measurement volume. The gratings are implemented as surface relief holograms in photoresist, which makes it possible to mass produce the beam splitter system at low cost through replication of the structure. The method for passive wavelength compensation for the fringe spacing is demonstrated both theoretically and experimentally. PMID- 14528926 TI - Realization of a multichannel integrated Young interferometer chemical sensor. AB - We report on the design, realization, and characterization of a four-channel integrated optical Young interferometer device that enables simultaneous and independent monitoring of three binding processes. The generated interference pattern is recorded by a CCD camera and analyzed with a fast-Fourier-transform algorithm. We present a thorough theoretical analysis of such a device. The realized device is tested by monitoring glucose solutions that induce well defined phase changes between output channels. The simultaneous measurement of three different glucose concentrations shows the multipurpose feature of such devices. The observed errors, caused by the mismatching of spatial frequencies of individual interference patterns with those determined from the CCD camera, are reduced with different reduction schemes. The phase resolution for different pairs of channels was approximately 1 x 10(-4) fringes, which corresponds to a refractive-index resolution of approximately 8.5 x 10(-8). The measured sensitivity coefficient of the phase change versus refractive-index change of approximately 1.22 x 10(3) x 2pi agrees well with the calculated coefficient of approximately 1.20 x 10(3) x 2pi. PMID- 14528927 TI - Measurement by multidirectional interferometers of the position and orientation of a positioning stage. AB - A multidirectional interferometer system is developed to determine the position and orientation of a stage moved in a two-dimensional (2-D) space. In this system four corner-cube prisms are mounted on the moving stage, and four laser beams are incident on the corner cubes in different directions. Moving distances in the observed directions are measured by laser interferometers. The position and orientation of the stage are calculated from the moving distances of the corner cubes. Some experiments are done on the 2-D moving stage with four interferometers, and measurement errors are estimated from redundant data. The estimated accuracy is higher than 0.2 microm for translation and 0.3 x 10(-3) deg for rotation for a measurement range of 0.5 mm and 0.5 deg. PMID- 14528928 TI - Increasing the range of unambiguity in step-height measurement with multiple wavelength interferometry--application to absolute long gauge block measurement. AB - An instrument for step-height measurement by multiple-wavelength interferometry is described. The addition of a 1152-nm wavelength to a multiple-wavelength scheme applying wavelengths of 633, 612, and 543 nm relaxes the tolerance range of the required preliminary measurement to +/- 140 microm, if the total uncertainty in the fringe fraction measurement can be kept below 2%. For larger fringe fraction measurement uncertainty, numerical simulations show that the integer number of interference orders can still be determined unambiguously if the range in the preliminary knowledge of the length has been correspondingly reduced. The interferometer instrument is described, and experimental data are presented in the context of long gauge block calibration at the National Research Council of Canada. PMID- 14528929 TI - Design criteria and optical characteristics of one-dimensional photonic crystals based on periodically grooved silicon. AB - Photonic bandgap (PBG) regions have been calculated for periodically grooved Si structures, acting as a one-dimensional photonic crystal. The wavelength range of the PBG as a function of the ratio (DSi/A) is presented, where DSi is the width of the Si walls and A is the grooved silicon lattice constant. The influence of the parameter DSi, the refractive index of the space between the Si walls and the number of structure periods, m, on the forming of PBG regions is discussed. A good correlation between the calculated and the experimentally observed PBG regions is obtained. PMID- 14528930 TI - Dichroic ultraviolet light filters. AB - With the intention to produce dichroic filters for use in photoluminescent systems that rely on polarized UV light, we synthesized a number of linear, dichroic dyes, which absorb mainly in the near-UV range of the electromagnetic spectrum. These dyes were designed for compatibility with common thermoplastic polymers such as linear low-density poly(ethylene), poly(ethylene terephthalate), and polyamide-12. Films of these host polymers that consisted of 0.2% by weight of various dichroic UV dyes were produced by common melt-processing schemes. Uniaxial drawing of these films yielded highly dichroic UV filters with dichroic ratios in absorption that in some cases exceeded 100. The fact that these free standing films display little or no coloration and are environmentally stable makes them useful for various applications that involve generation of polarized UV light. PMID- 14528932 TI - Lens based on the use of left-handed materials. AB - A new lens that is a modification of the Veselago lens is proposed. It consists of a slab of the left-handed material embedded into a regular material. The materials for the new lens should have the same refractive index, unlike that of the Veselago lens in which the materials should in addition have the same impedance. Therefore the new lens should be easier to manufacture. As the Veselago lens, the new lens might be useful for the three dimensional imaging. In contrast to the Veselago lens the new lens has multiple foci, and it may image an object that is located at any large distance from the slab. PMID- 14528931 TI - Three-dimensional imaging of microspheres with confocal and conventional polarization microscopes. AB - We experimentally studied the three-dimensional imaging of the microspheres by using confocal and conventional scanning polarization microscopes. Because of the field amplitude averaging effect of the confocal system, the polarization of the detected signals is mainly parallel to the initial polarization. As a result, the signal intensity from the microspheres in the confocal polarization microscope with a crossed analyzer was found to be weaker than that in the conventional system. Based on a vector approach that takes the polarization into account and on the image formations of the two systems, theoretical expressions are given that agree well with the experimental results. PMID- 14528933 TI - Wet-etch figuring for precision optical contouring. AB - Wet-etch figuring utilizes free surface flows driven by surface tension gradients (the Marangoni effect) to confine and stabilize the size and shape of an etchant droplet attached to the underside of a glass surface. This droplet, or wetted zone, is translated on the surface, etching where it contacts and leaving behind no residue, to facilitate an etching-based small-tool figuring process that is free of mechanical and thermal stresses. The optic needs no backing plate, and its back side is free for inspection by optical means. When transmissive optics is figured, the optical thickness between the front and the rear surfaces of the optic is measured interferometrically and used in real time to control the local dwell time of the etchant zone. This truly closed-loop figuring process is robust, environmentally insensitive, and fully automated. It is particularly suited for figuring patterns such as phase plates, corrective elements, and optical flats on very thin (<< 1-mm) substrates that are difficult to figure with traditional abrasive polishing methods. PMID- 14528934 TI - Optical resonances in periodic surface arrays of metallic patches. AB - The transmission of light along the surface normal through an air-quartz-glass interface covered with a periodic array of thin, rectangular gold patches has been studied over the visible to infrared range. The various structures that are observed can be qualitatively understood as arising from standing-wave resonances set by the size and surroundings of the metal patches. A method-of-moments calculational scheme provides simulations in good quantitative agreement with the data. It is shown how the standing-wave picture provides a useful conceptual framework to understand and exploit such systems. PMID- 14528935 TI - Polarization control by use of the electro-optic effect in periodically poled lithium niobate. AB - We propose a method to control the polarization of light by the electro-optic effect in periodically poled lithium niobate. A single integrated chip of Z-cut lithium niobate having two sections is used. The first section is not periodically poled, whereas the second section is. With an electric field applied along the Z axis of the first section and another electric field applied along the Y axis of the second section, light with an arbitrary elliptical polarization can be converted into a fixed linearly polarized state. PMID- 14528936 TI - Theoretical model of the polarization properties of the retinal nerve fiber layer in reflection. AB - In several optical technologies for glaucoma diagnosis, polarized light is used to assess the retinal nerve fiber layer (RNFL) of the eye. For better understanding of the polarization properties of the RNFL, it was modeled as a thick birefringent slab containing parallel light-scattering cylinders, and the Mueller matrix for reflectance was derived. The model predicts that (1) the RNFL reflectance has weak intrinsic diattenuation; (2) the diattenuation spectrum depends strongly on the relative refractive indices of the cylinders; (3) both scattering and birefringence contribute to retardation; and (4) the RNFL reflectance generally preserves polarization, but depolarization may be detectable for thick RNFL at short wavelengths. PMID- 14528938 TI - Spatial pattern separation of chemicals and frequency-independent components by terahertz spectroscopic imaging. AB - We separated the component spatial patterns of frequency-dependent absorption in chemicals and frequency-independent components such as plastic, paper, and measurement noise in terahertz (THz) spectroscopic images, using known spectral curves. Our measurement system, which uses a widely tunable coherent THz-wave parametric oscillator source, can image at a specific frequency in the range 1-2 THz. The component patterns of chemicals can easily be extracted by use of the frequency-independent components. This method could be successfully used for nondestructive inspection for the detection of illegal drugs and devices of bioterrorism concealed, e.g., inside mail and packages. PMID- 14528937 TI - Diattenuation and polarization preservation of retinal nerve fiber layer reflectance. AB - The diattenuation spectrum of the retinal nerve fiber layer (RNFL) reflectance has been predicted to depend strongly on the relative refractive index (m) of light-scattering cylinders. To constrain the values of m, diattenuation of the RNFL reflectance of isolated rat retina was measured with a multispectral imaging micropolarimeter. The RNFL reflection has very weak intrinsic diattenuation at all wavelengths (400-830 nm), which rejects all values of m > or = 1.03. Degree of polarization (DOP) for reflection from the RNFL was also measured. DOP was close to unity at all wavelengths, which indicates that the RNFL is a polarization-preserving reflector. PMID- 14528939 TI - Design and performance of capping layers for extreme-ultraviolet multilayer mirrors. AB - Multilayer lifetime has emerged as one of the major issues for the commercialization of extreme-ultraviolet lithography (EUVL). We describe the performance of an oxidation-resistant capping layer of Ru atop multilayers that results in a reflectivity above 69% at 13.2 nm, which is suitable for EUVL projection optics and has been tested with accelerated electron-beam and extreme ultraviolet (EUV) light in a water-vapor environment. Based on accelerated exposure results, we calculated multilayer lifetimes for all reflective mirrors in a typical commercial EUVL tool and concluded that Ru-capped multilayers have approximately 40x longer lifetimes than Si-capped multilayers, which translates to 3 months to many years, depending on the mirror dose. PMID- 14528940 TI - Efficient characterization of regional mesenteric blood flow by use of laser speckle imaging. AB - We present a noninvasive full-field method--laser speckle imaging (LSI)--for measuring the regional mesenteric blood flow without scanning. A system of LSI was designed and validated in a model experiment. Dynamics of regional blood flow in the rat mesentery under the influence of noradrenaline were monitored by this method. Spatial and temporal characteristics of the mesenteric blood-flow response were achieved with high resolution. These suggested that LSI might provide a new approach to microcirculation studies. PMID- 14528941 TI - Optical forces for noninvasive cellular analysis. AB - A novel, noninvasive measurement technique for quantitative cellular analysis is presented that utilizes the forces generated by an optical beam to evaluate the physical properties of live cells in suspension. In this analysis, a focused, near-infrared laser line with a high cross-sectional intensity gradient is rapidly scanned across a field of cells, and the interaction of those cells with the beam is monitored. The response of each cell to the laser depends on its size, structure, morphology, composition, and surface membrane properties; therefore, with this technique, cell populations of different type, treatment, or biological state can be compared. To demonstrate the utility of this cell analysis platform, we evaluated the early stages of apoptosis induced in the U937 cancer cell line by the drug camptothecin and compared the results with established reference assays. Measurements on our platform show detection of cellular changes earlier than either of the fluorescence-based Annexin V or caspase assays. Because no labeling or additional cell processing is required and because accurate assays can be performed with a small number of cells, this measurement technique may find suitable applications in cell research, medical diagnostics, and drug discovery. PMID- 14528942 TI - Interferometric optical sensor for measuring glucose concentration. AB - With a specially designed probe, the phase difference between s andp polarization of light reflected under surface-plasmon resonance is measured by use of a common path heterodyne interferometer. For specific ratios of phase difference to glucose concentration, the glucose concentration can be estimated as a function of the measured phase data. A prototype was set up to demonstrate the feasibility of this sensor, which was experimentally tested in the range 40-500 mg/dl with a small quantity of solution and had a measurement resolution of 1.41 mg/dl at 25 degrees C. PMID- 14528943 TI - Measurement and analysis of light distribution in intralipid-10% at 650 nm. AB - A method for measuring the distributions of light in the biological tissue phantom Intralipid by use of an optical fiber is presented and measurements of distributions of light in Intralipid-10% suspensions at 650 nm are described. This approach is complementary to that in which an optical fiber with an isotropic tip detects the distribution of light in tissue phantoms. The characteristics of the distance-dependent intensity of scattering light in different directions were revealed by the experimental results; the effects of the optical parameters and of the radius of the incident beam on the distributions of light in tissue phantoms were given; and the experimental results were analyzed by the diffusion theory. These studies will help in further understanding of the scattering characteristics of tissue. PMID- 14528944 TI - Evaluation of optical properties of highly scattering media by moments of distributions of times of flight of photons. AB - A novel method for the determination of the optical properties of tissue from time-domain measurements is presented. The data analysis is based on the evaluation of the first moment and the second centralized moment, i.e., the mean time of flight and the variance of the measured distribution of times of flight (DTOF) of photons injected by short (picosecond) laser pulses. Analytical expressions are derived for calculation of absorption and of reduced scattering coefficients from these moments by application of diffusion theory for infinite and semi-infinite homogeneous media. The proposed method was tested on experimental data obtained with phantoms, and results for absorption and reduced scattering coefficients obtained by the proposed method are compared with those obtained by fitting of the same data with analytical solutions of the diffusion equation. Furthermore, the accuracy of the moment analysis was investigated for a range of integration limits of the DTOF. The moment analysis may serve as a comparatively fast method for evaluating optical properties with sufficient accuracy and can be used, e.g., for on-line monitoring of optical properties of biological tissue. PMID- 14528945 TI - It is necessary to validate each individual IMRT treatment plan before delivery. For the proposition. PMID- 14528946 TI - It is necessary to validate each individual IMRT treatment plan before delivery. Against the proposition. PMID- 14528947 TI - A new algorithm for the registration of portal images to planning images in the verification of radiotherapy, as validated in prostate treatments. AB - The manual verification of a radiotherapy treatment, where a portal image is matched onto a planning image, is very time consuming and subject to inter- and intraobserver variability. Therefore, a fully automatic matching procedure (image registration) is required. Existing automatic matching algorithms are confounded, however, by irrelevant information in the portal images (i.e., air in the intestines). Therefore, we have developed a new method, which is an extension of chamfer matching and uses, apart from the distance to the nearest edge, additional information on the correspondence of the gradient angle and magnitude of the edges, making the method less sensitive to confounding information in the images. To validate the automatic matching procedure in clinical practice, we applied the new method on 157 images of 29 randomly selected patients treated for carcinoma of the prostate. Three experts manually matched these images in consensus. Subsequently, the same observers assessed the results of the automatic registration. When regular chamfer matching is used for the fully automatic matching procedure, only 5% of the image pairs could be matched correctly, whereas the new method successfully registered 80% by using additional information on the angle of the edges. From the results of the validation study it can be concluded that a significant reduction in workload for the physicians and technicians can be achieved with this method. PMID- 14528948 TI - Effect of saccharide additives on response of ferrous-agarose-xylenol orange radiotherapy gel dosimeters. AB - Glucose, sucrose, starch, and locust bean gum have been used as additives to the ferrous-agarose-xylenol orange (FAX) gel dosimeter. The saccharide enhanced dosimeters were found to have a higher dose sensitivity over a standard FAX gel as measured by both optical density change and magnetic resonance imaging (MRI). With optical density measurement, OD-dose sensitivity increases were up to 55% for glucose, 122% for sucrose and 43% for starch, while locust bean gum did not give a consistent response. With MRI, R1-dose sensitivity increases were up to 178% with sucrose addition. The FAX gel with sucrose was studied in greatest detail. The OD-dose sensitivity dependence on cooling rate was reduced for the sucrose FAX gel over the standard FAX gel, which has significant implications for uniform dose sensitivity in large gel phantoms. The thermal oxidation rate in the sucrose FAX gel was up to 2.3 times higher than in the standard gel. The OD-dose sensitivity of oxygenated sucrose FAX gels was 4.3 times greater than standard FAX gels, while continued enhancement in OD-dose sensitivity with increased sucrose concentrations beyond 2.0 g/l was found only for the oxygenated sucrose FAX gels. Both the molar absorption coefficient of the ferric ion-xylenol orange complex at 543 nm and gel pH were not affected by the presence of sucrose, with the implication that the higher OD-dose sensitivity of gels with saccharides is due to increased chain reaction production of ferric ions. PMID- 14528949 TI - Dual focal-spot imaging for phase extraction in phase-contrast radiography. AB - The purpose of this study was to evaluate dual focal spot imaging as a method for extracting the phase component from a phase-contrast radiography image. All measurements were performed using a microfocus tungsten-target x-ray tube with an adjustable focal-spot size (0.01 mm to 0.045 mm). For each object, high resolution digital radiographs were obtained with two different focal spot sizes to produce matched image pairs in which all other geometric variables as well as total exposure and tube kVp were held constant. For each image pair, a phase extraction was performed using pixel-wise division. The phase-extracted image resulted in an image similar to the standard image processing tool commonly referred to as "unsharp masking" but with the additional edge-enhancement produced by phase-contrast effects. The phase-extracted image illustrates the differences between the two images whose imaging parameters differ only in focal spot size. The resulting image shows effects from both phase contrast as well as geometric unsharpness. In weakly attenuating materials the phase-contrast effect predominates, while in strongly attenuating materials the phase effects are so small that they are not detectable. The phase-extracted image in the strongly attenuating object reflects differences in geometric unsharpness. The degree of phase extraction depends strongly on the size of the smallest focal spot used. This technique of dual-focal spot phase-contrast radiography provides a simple technique for phase-component (edge) extraction in phase-contrast radiography. In strongly attenuating materials the phase-component is overwhelmed by differences in geometric unsharpness. In these cases the technique provides a form of unsharp masking which also accentuates the edges. Thus, the two effects are complimentary and may be useful in the detection of small objects. PMID- 14528950 TI - On the use of C-arm fluoroscopy for treatment planning in high dose rate brachytherapy. AB - Treatment planning for brachytherapy requires the acquisition of geometrical information of the implant applicator and the patient anatomy. This is typically done using a simulator or a computed tomography scanner. In this study, we present a different method by which orthogonal images from a C-arm fluoroscopic machine is used for high dose rate brachytherapy treatment planning. A typical C arm is not isocentric, and it does not have the mechanical accuracy of a simulator. One solution is to place a reconstruction box with fiducial markers around the patient. However, with the limited clearance of the C-arm this method is very cumbersome to use, and is not suitable for all patients and implant sites. A different approach is adopted in our study. First, the C-arm movements are limited to three directions only between the two orthogonal images: the C orbital rotation, the vertical column, and the horizontal arm directions. The amounts of the two linear movements and the geometric parameters of the C-arm orbit are used to calculate the location of the crossing point of the two beams and thus the magnification factors of the two images. Second, the fluoroscopic images from the C-arm workstation are transferred in DICOM format to the planning computer through a local area network. Distortions in the fluoroscopic images, with its major component the "pincushion" effect, are numerically removed using a software program developed in house, which employs a seven-parameter polynomial filter. The overall reconstruction accuracy using this method is found to be 2 mm. This filmless process reduces the overall time needed for treatment planning, and greatly improves the workflow for high dose rate brachytherapy procedures. Since its commissioning nearly three years ago, this system has been used extensively at our institution for endobronchial, intracavitary, and interstitial brachytherapy planning with satisfactory results. PMID- 14528952 TI - ROC analysis for assessment of lesion detection performance in 3D PET: influence of reconstruction algorithms. AB - Image quality in positron emission tomography (PET) can be assessed with physical parameters, as spatial resolution and signal-to-noise ratio, or using psychophysical approaches, which include the observer performance and the considered task (ROC analysis). For PET in oncology, such a task is the detection of hot lesions. The aim of the present study was to assess the lesion detection performance due to adequate modeling of the scanner and the measurement process in the image reconstruction process. We compared the standard OSEM software of the manufacturer with a sophisticated fully 3D iterative reconstruction technique (USC MAP). A rectangular phantom with 6 oblique line sources in a homogeneous background (2.6 kBq/ml 18F) was imaged dynamically with an ECAT EXACT HR+ scanner in 3D mode. Reconstructed activity contrasts varied between 15 and 0, as the line sources were filled with 11C (3.2 MBq/ml). Measured attenuation and standard randoms, dead time, and scatter corrections of the manufacturer were employed. For the ROC analysis, a software tool presented a cut-out of the phantom (15 x 15 pixels) to two observers. These cut-outs were rated (5 classes) and the area Az under the ROC curve was determined as a measure of detection performance. The improvement for Az with USC MAP compared to the OSEM reconstructions ranged between 0.02 and 0.23 for signal-to-noise ratios of the background between 2.8 and 3.1 and lesion contrast between 2.1 and 4.2. This study demonstrates that adequate modeling of the measurement process in the reconstruction algorithm improves the detection of small hot lesions markedly. PMID- 14528951 TI - A stereotactic method for the three-dimensional registration of multi-modality biologic images in animals: NMR, PET, histology, and autoradiography. AB - The objective of this work was to develop and then validate a stereotactic fiduciary marker system for tumor xenografts in rodents which could be used to co register magnetic resonance imaging (MRI), PET, tissue histology, autoradiography, and measurements from physiologic probes. A Teflon fiduciary template has been designed which allows the precise insertion of small hollow Teflon rods (0.71 mm diameter) into a tumor. These rods can be visualized by MRI and PET as well as by histology and autoradiography on tissue sections. The methodology has been applied and tested on a rigid phantom, on tissue phantom material, and finally on tumor bearing mice. Image registration has been performed between the MRI and PET images for the rigid Teflon phantom and among MRI, digitized microscopy images of tissue histology, and autoradiograms for both tissue phantom and tumor-bearing mice. A registration accuracy, expressed as the average Euclidean distance between the centers of three fiduciary markers among the registered image sets, of 0.2 +/- 0.06 mm was achieved between MRI and microPET image sets of a rigid Teflon phantom. The fiduciary template allows digitized tissue sections to be co-registered with three-dimensional MRI images with an average accuracy of 0.21 and 0.25 mm for the tissue phantoms and tumor xenografts, respectively. Between histology and autoradiograms, it was 0.19 and 0.21 mm for tissue phantoms and tumor xenografts, respectively. The fiduciary marker system provides a coordinate system with which to correlate information from multiple image types, on a voxel-by-voxel basis, with sub-millimeter accuracy--even among imaging modalities with widely disparate spatial resolution and in the absence of identifiable anatomic landmarks. PMID- 14528953 TI - A toolbox for intensity modulated radiation therapy optimization. AB - We have designed a toolbox that provides an environment for testing radiotherapy optimization techniques, objective functions, and constraints. A set of three dimensional (3D) pencil beam dose distributions have been computed for a cylindrical phantom. The 6 MV pencil beams were computed using a superposition based dose engine commissioned for an Elekta SL20 linear accelerator. Due to the cylindrical symmetry of the phantom, the pencil beam dose distributions for any arbitrary beam angle can be determined by simply rotating the pencil beam data sets. Thus, the full accuracy is maintained without the need for additional dose calculations or large data storage requirements. In addition to the pencil beam data sets, tools are included for (1) rotating the pencil beams, (2) calculating the beam's eye view, (3) drawing structures, (4) writing the pencil beam dose data out to the optimizer, and (5) visualizing the optimized results. The pencil beam data sets and the corresponding tools are available for download at http://medschool.umaryland.edu/departments/radiationoncology/pencilbeam/. With this toolbox, researchers will have the ability to rapidly test new optimization techniques and formulations for intensity modulated radiation therapy and 3D conformal radiotherapy. PMID- 14528954 TI - Accuracy of a simple method for deriving the presampled modulation transfer function of a digital radiographic system from an edge image. AB - Several methods for accurately deriving the presampled modulation transfer function (MTF) of a pixelated detector from the image of a slightly slanted edge have been described in the literature. In this paper we report on a simple variant of the edge method that produces sufficiently accurate MTF values for frequencies up to the Nyquist frequency limit of the detector with little effort in edge alignment and computation. The oversampled ESF is constructed in a very simple manner by rearranging the pixel data of N consecutive lines corresponding to a lateral shift of the edge by one pixel. A regular subsampling pitch is assumed for the oversampled ESF, which is given by the original pixel sampling distance divided by the integer number N. This allows the original data to be used for further computational analysis (differentiation and Fourier transform) without data preprocessing. Since the number of lines leading to an edge shift by one pixel generally is a fractional number rather than an integer, a systematic error may be introduced into the presampled MTF. Simulations and theoretical investigations show that this error is proportional to 1/N and increases with spatial frequency. For all frequencies up to the Nyquist limit, the relative error delta MTF/MTF is smaller than 1/(2N). It can thus be kept below a given threshold by suitably selecting N, which furnishes a certain maximum edge angle. The method is especially useful for applications where the presampled MTF is needed only for frequencies up to the Nyquist frequency limit, such as the determination of the detective quantum efficiency (DQE). PMID- 14528955 TI - From physical dose constraints to equivalent uniform dose constraints in inverse radiotherapy planning. AB - Optimization algorithms in inverse radiotherapy planning need information about the desired dose distribution. Usually the planner defines physical dose constraints for each structure of the treatment plan, either in form of minimum and maximum doses or as dose-volume constraints. The concept of equivalent uniform dose (EUD) was designed to describe dose distributions with a higher clinical relevance. In this paper, we present a method to consider the EUD as an optimization constraint by using the method of projections onto convex sets (POCS). In each iteration of the optimization loop, for the actual dose distribution of an organ that violates an EUD constraint a new dose distribution is calculated that satisfies the EUD constraint, leading to voxel-based physical dose constraints. The new dose distribution is found by projecting the current one onto the convex set of all dose distributions fulfilling the EUD constraint. The algorithm is easy to integrate into existing inverse planning systems, and it allows the planner to choose between physical and EUD constraints separately for each structure. A clinical case of a head and neck tumor is optimized using three different sets of constraints: physical constraints for all structures, physical constraints for the target and EUD constraints for the organs at risk, and EUD constraints for all structures. The results show that the POCS method converges stable and given EUD constraints are reached closely. PMID- 14528956 TI - Analytic modeling of breast elastography. AB - The elastic moduli of tumors change during their pathological evolution. Elastographic imaging has potential for detecting and characterizing cancers by mapping the stiffness distribution in tissues. In this paper a micromechanics based analytical method was developed to detect the location, size, and elastic modulus of a tumor mass embedded in a symmetric two-dimensional breast tissue. A closed-form solution for the strain elastograms (forward problem) was derived. A computational algorithm for the inverse problem was developed for the detection, localization, and characterization of a heterogeneous mass embedded in a breast tissue. Numerical examples were presented to evaluate the proposed method's performance. The detectability of a tumor mass was estimated with respect to lesion location, size, and modulus contrast ratio. It was shown that the micromechanics theory provides a powerful tool for the diagnosis of breast cancer. PMID- 14528957 TI - Improved artificial neural networks in prediction of malignancy of lesions in contrast-enhanced MR-mammography. AB - The aim of this study was to evaluate the capability of improved artificial neural networks (ANN) and additional novel training methods in distinguishing between benign and malignant breast lesions in contrast-enhanced magnetic resonance-mammography (MRM). A total of 604 histologically proven cases of contrast-enhanced lesions of the female breast at MRI were analyzed. Morphological, dynamic and clinical parameters were collected and stored in a database. The data set was divided into several groups using random or experimental methods [Training & Testing (T&T) algorithm] to train and test different ANNs. An additional novel computer program for input variable selection was applied. Sensitivity and specificity were calculated and compared with a statistical method and an expert radiologist. After optimization of the distribution of cases among the training and testing sets by the T & T algorithm and the reduction of input variables by the Input Selection procedure a highly sophisticated ANN achieved a sensitivity of 93.6% and a specificity of 91.9% in predicting malignancy of lesions within an independent prediction sample set. The best statistical method reached a sensitivity of 90.5% and a specificity of 68.9%. An expert radiologist performed better than the statistical method but worse than the ANN (sensitivity 92.1%, specificity 85.6%). Features extracted out of dynamic contrast-enhanced MRM and additional clinical data can be successfully analyzed by advanced ANNs. The quality of the resulting network strongly depends on the training methods, which are improved by the use of novel training tools. The best results of an improved ANN outperform expert radiologists. PMID- 14528958 TI - Beam orientation optimization for IMRT by a hybrid method of the genetic algorithm and the simulated dynamics. AB - We have developed a new method for beam orientation optimization in intensity modulated radiation therapy (IMRT). The problem of beam orientation optimization in IMRT is solved by a decoupled two-step iterative process: (1) optimization of the intensity profiles for given beam configurations; (2) selection of optimal beam configurations based on the ranking by an objective function score for the results of the intensity profile optimization. The simulated dynamics algorithm is used for the intensity profile optimization. This algorithm enforces both the hard constraints and dose-volume constraints. A genetic algorithm is used to select beam orientation configurations. The method has been tested for both a simulated and clinical case, and the results show that beam orientation optimization significantly improved IMRT plans within a time period that is clinically acceptable. The results also show the dependence of the optimal orientation configurations on the prescribed constraints. In addition, beam orientation optimization by the method described here can provide multiple plans with similar dose distributions. This degeneracy characteristic can be exploited to our advantage in introducing additional planning objectives, e.g., the smoothness of intensity profiles, for the selection of the optimal plan among the degenerate configurations for treatment delivery. PMID- 14528959 TI - Are dose calculations during dose optimization in brachytherapy necessary? AB - We consider the problem of anatomy based dose optimization in brachytherapy. A calculation method for some objective functions and their derivatives is proposed which significantly reduces the number of required operations. The optimization in some cases, ignoring a preprocessing step, is independent of the number of sampling points. The idea is that some of the objectives and their derivatives used for dose optimization do not require the explicit calculation of dose values. Dose optimization with the new modified computation method for the objectives and derivatives is, depending on the number of sampling points, up to 100 times faster than the conventional method with dose calculation. PMID- 14528960 TI - Three-dimensional dose reconstruction of breast cancer treatment using portal imaging. AB - In this study, we present an algorithm for three-dimensional (3-D) dose reconstruction using portal images obtained with an electronic portal imaging device (EPID). For this purpose an algorithm for 2-D dose reconstruction, which was previously developed in our institution, was adapted. The external contour of the patient was used to correct for absorption of primary photons, but the presence of inhomogeneities was not taken into account. The accuracy of the algorithm was determined by irradiating two anthropomorphic breast phantoms with 6 MV photons. The dose values derived from portal images were compared with results from 3-D dose calculations, which, in turn, were verified with data obtained with an ionization chamber and film dosimetry. It was found that the application of contour information significantly improves the accuracy of 2-D dose reconstruction. If the total dose at the isocenter plane resulting from all treatment beams is reconstructed, the average deviation from the planned dose is 0.1%+/-1.7% (1 SD). If contour information is not available, the differences increase up to +/-20% for the individual beams. In that case, the dose can only be reconstructed with reasonable accuracy when (nearly) opposing beams are used. The average deviation of the 3-D reconstructed dose from the planned dose in the irradiated volume is 1.4%+/-5.4% (1 SD). If the irradiated volume is enclosed by planes less than 5 cm distant from the isocenter plane, then the average deviation is only 0.5%+/-3.4% (1 SD). It can be concluded that the proposed algorithm for a 3-D dose reconstruction allows a determination of the dose at the isocenter plane and the dose-volume histogram with an accuracy acceptable for an independent verification of the treatment. PMID- 14528962 TI - High precision film dosimetry with GAFCHROMIC films for quality assurance especially when using small fields. AB - Treatment units for radiosurgery, brachytherapy, implementation of seeds, and IMRT generate small high dose regions together with steep dose gradients of up to 30%-50% per mm. Such devices are used to treat small complex-shaped lesions, often located close to critical structures, by superimposing several single high dose regions. In order to test and verify these treatment techniques, to perform quality assurance tasks and to simulate treatment conditions as well as to collect input data for treatment planning, a GAFCHROMIC film based dosimetry system for measuring two-dimensional (2-D) and three-dimensional (3-D) dose distributions was developed. The nearly tissue-equivalent radiochromic GAFCHROMIC film was used to measure dose distributions. A drum scanner was investigated and modified. The spectral emission of the light source and the filters together with the efficiency of the CCD filters for the red color were matched and balanced with the absorption spectra of the film. Models based on refined studies have been developed to characterize theoretically the physics of film exposure and to calibrate the film. Mathematical descriptions are given to calculate optical densities from spectral data. The effect of darkening has been investigated and is described with a mathematical model. The influence of the scan temperature has been observed and described. In order to cope with the problem of individual film inhomogeneities, a double irradiation technique is introduced and implemented that yields dose accuracies as good as 2%-3%. Special software routines have been implemented for evaluating and handling the film data. PMID- 14528961 TI - Detection of prostate cancer by integration of line-scan diffusion, T2-mapping and T2-weighted magnetic resonance imaging; a multichannel statistical classifier. AB - A multichannel statistical classifier for detecting prostate cancer was developed and validated by combining information from three different magnetic resonance (MR) methodologies: T2-weighted, T2-mapping, and line scan diffusion imaging (LSDI). From these MR sequences, four different sets of image intensities were obtained: T2-weighted (T2W) from T2-weighted imaging, Apparent Diffusion Coefficient (ADC) from LSDI, and proton density (PD) and T2 (T2 Map) from T2 mapping imaging. Manually segmented tumor labels from a radiologist, which were validated by biopsy results, served as tumor "ground truth." Textural features were extracted from the images using co-occurrence matrix (CM) and discrete cosine transform (DCT). Anatomical location of voxels was described by a cylindrical coordinate system. A statistical jack-knife approach was used to evaluate our classifiers. Single-channel maximum likelihood (ML) classifiers were based on 1 of the 4 basic image intensities. Our multichannel classifiers: support vector machine (SVM) and Fisher linear discriminant (FLD), utilized five different sets of derived features. Each classifier generated a summary statistical map that indicated tumor likelihood in the peripheral zone (PZ) of the prostate gland. To assess classifier accuracy, the average areas under the receiver operator characteristic (ROC) curves over all subjects were compared. Our best FLD classifier achieved an average ROC area of 0.839(+/-0.064), and our best SVM classifier achieved an average ROC area of 0.761(+/-0.043). The T2W ML classifier, our best single-channel classifier, only achieved an average ROC area of 0.599(+/-0.146). Compared to the best single-channel ML classifier, our best multichannel FLD and SVM classifiers have statistically superior ROC performance (P=0.0003 and 0.0017, respectively) from pairwise two-sided t-test. By integrating the information from multiple images and capturing the textural and anatomical features in tumor areas, summary statistical maps can potentially aid in image-guided prostate biopsy and assist in guiding and controlling delivery of localized therapy under image guidance. PMID- 14528963 TI - Quantitative imaging of bromine-76 and yttrium-86 with PET: a method for the removal of spurious activity introduced by cascade gamma rays. AB - Positron Emission Tomography of bromine-76 and yttrium-86 results in the detection of coincident events that are not strictly associated with annihilation photon pairs. Instead, these coincidences occur because prompt gamma rays emitted by these nuclides result in cascades of photons that are emitted within the timing window of the PET scanner. Pairs of detected photons from these cascades are not angularly correlated and therefore contain little information regarding the location of their source. Furthermore, these coincidences are not removed by correction procedures (e.g., randoms, scatter) routinely applied to PET data. If left uncorrected, the cascade coincidences will result in spurious apparent activity within the PET images. A correction, applied within projection space, that removes the cascade coincidence signal from septa-in (i.e., two-dimensional) datasets is proposed and tested on phantom data. PMID- 14528964 TI - Dosimetric characteristics of a low-kV intra-operative x-ray source: implications for use in a clinical trial for treatment of low-risk breast cancer. AB - The dosimetric characteristics of a low-kV x-ray device for performing intra operative irradiations, the Intrabeam (Photoelectron Corporation, Lexington, MA), are examined. Two dosimetric models are considered--an analytical model considering only the primary x-ray beam, and a Monte Carlo model utilizing the EGSnrc code and a spherical simulation geometry. Both models prove reliable for verifying measured dose distributions for the device. The Monte Carlo model is necessary for examining spectral variations and the influence of inhomogeneities. The predictions of the Monte Carlo model are utilized to examine points of consideration for a multi-center clinical trial using the Intrabeam in the intra operative, single fraction post-resection treatment of low-risk breast cancer. Predicted differences in radiological equivalence of breast tissue and water suggest a 3-5% under-dose of breast tissue (in a 50 kV beam) when dose fall-off data in water is used. A substantial dose enhancement in bone (i.e., ribs) adjacent to the treatment site is predicted though, based on published clinical data for radiation-induced rib fracture, it is concluded that induction of radiation-induced rib fracture would not pose a significant risk. Dose-volume changes with size of the treatment area (defined by the size of the resection volume) are examined indicating large variations in dose-volume characteristics across the range of possible "target" volumes. PMID- 14528965 TI - Analysis of various beamlet sizes for IMRT with 6 MV photons. AB - Application of intensity modulated radiation therapy (IMRT) using multileaf collimation often requires the use of small beamlets to optimize the delivered radiation distribution. Small-beam dose distribution measurements were compared to dose distributions calculated using a commercial treatment planning system that models its data acquired using measurements from relatively large fields. We wanted to evaluate only the penumbra, percent depth-dose (PDD) and output model, so we avoided dose distribution features caused by rounded leaf ends and interleaf leakage by making measurements using the secondary collimators. We used a validated radiochromic film dosimetry system to measure high-resolution dose distributions of 6 MV photon beams. A commercial treatment planning system using the finite size pencil beam (FSPB) dose calculation algorithm was commissioned using measured central axis outputs from 4.0x4.0 to 40.0x40.0 cm2 beams and radiographic-film profile measurements of a 4.0x4.0 cm2 beam at twice the depth of maximum dose (dmax). Calculated dose distributions for square fields of 0.5x0.5 cm2, and 1.0x1.0 cm2, to 6.0x6.0 cm2, in 1.0x1.0 cm2, increments were compared against radiochromic film measurements taken with the film oriented parallel to the beam central axis in a water equivalent phantom. The PDD of the smaller field sizes exhibited behavior typical of small fields, namely a decrease in dmax with decreasing field size. The FSPB accurately modeled the depth-dose and central axis output for depths deeper than the nominal dmax of 1.5 cm plus 0.5 cm. The dose distribution in the build-up and penumbra regions was not accurately modeled for depths less than 2 cm, especially for the fields of 2.0x2.0 cm2 and smaller. Using the gamma function with 2 mm and 2% criteria, the dose model was shown to accurately predict the penumbra. While for single small beams the compared dose distributions passed the gamma function criteria, the clinical appropriateness of these criteria is not clear for a composite IMRT plan. Further investigation of the cumulative impact of the observed dose discrepancies is warranted. We speculate that the observed differences in the penumbra regions arise from some energy dependent artifact in the radiographic film profiles used for commissioning. In the future, radiochromic film based commissioning might provide a more accurate data set for dose modeling. PMID- 14528966 TI - Quantitative computerized analysis of diffuse lung disease in high-resolution computed tomography. AB - An automated computerized scheme has been developed for the detection and characterization of diffuse lung diseases on high-resolution computed tomography (HRCT) images. Our database consisted of 315 HRCT images selected from 105 patients, which included normal and abnormal slices related to six different patterns, i.e., ground-glass opacities, reticular and linear opacities, nodular opacities, honeycombing, emphysematous change, and consolidation. The areas that included specific diffuse patterns in 315 HRCT images were marked by three radiologists independently on the CRT monitor in the same manner as they commonly describe in their radiologic reports. The areas with a specific pattern, which three radiologists marked independently and consistently as the same patterns, were used as "gold standard" for specific abnormal opacities in this study. The lungs were first segmented from the background in each slice by use of a morphological filter and a thresholding technique, and then divided into many contiguous regions of interest (ROIs) with a 32x32 matrix. Six physical measures which were determined in each ROI included the mean and the standard deviation of the CT value, air density components, nodular components, line components, and multilocular components. Artificial neural networks (ANNs) were employed for distinguishing between seven different patterns which included normals and six patterns associated with diffuse lung disease. The sensitivity of this computerized method for a detection of the six abnormal patterns in each ROI was 99.2% (122/123) for ground-glass opacities, 100% (15/15) for reticular and linear opacities, 88.0% (132/150) for nodular opacities, 100% (98/98) for honeycombing, 95.8% (369/385) for emphysematous change, and 100% (43/43) for consolidation. The specificity in detecting a normal ROI was 88.1% (940/1067). This computerized method may be useful in assisting radiologists in their assessment of diffuse lung disease in HRCT images. PMID- 14528967 TI - Evaluation of the gamma dose distribution comparison method. AB - The gamma tool was developed to quantitatively compare dose distributions, either measured or calculated. Before computing gamma, the dose and distance scales of the two distributions, referred to as evaluated and reference, are renormalized by dose and distance criteria, respectively. The renormalization allows the dose distribution comparison to be conducted simultaneously along dose and distance axes. The gamma quantity, calculated independently for each reference point, is the minimum distance in the renormalized multidimensional space between the evaluated distribution and the reference point. The gamma quantity degenerates to the dose-difference and distance-to-agreement tests in shallow and very steep dose gradient regions, respectively. Since being introduced, the gamma quantity has been used by investigators to evaluate dose calculation algorithms, and compare dosimetry measurements. This manuscript examines the gamma distribution behavior in two dimensions and evaluates the gamma distribution in the presence of data noise. Noise in the evaluated distribution causes the gamma distribution to be underestimated relative to the no-noise, condition. Noise in the reference distribution adds noise in the gamma distribution in proportion to the normalized dose noise. In typical clinical use, the fraction of points that exceed 3% and 3 mm can be extensive, so we typically use 5% and 2-3 mm in clinical evaluations. For clinical cases, the calculation time is typically 5 minutes for a 1 x 1 mm2 interpolated resolution on an 800 MHz Pentium 4 for a 14.1 x 15.2 cm2 radiographic film. PMID- 14528968 TI - A reconstruction algorithm for coherent scatter computed tomography based on filtered back-projection. AB - Coherent scatter computed tomography (CSCT) is a reconstructive x-ray imaging technique that yields the spatially resolved coherent-scatter form factor of the investigated object. Reconstruction from coherently scattered x-rays is commonly done using algebraic reconstruction techniques (ART). In this paper, we propose an alternative approach based on filtered back-projection. For the first time, a three-dimensional (3D) filtered back-projection technique using curved 3D back projection lines is applied to two-dimensional coherent scatter projection data. The proposed algorithm is tested with simulated projection data as well as with projection data acquired with a demonstrator setup similar to a multi-line CT scanner geometry. While yielding comparable image quality as ART reconstruction, the modified 3D filtered back-projection algorithm is about two orders of magnitude faster. In contrast to iterative reconstruction schemes, it has the advantage that subfield-of-view reconstruction becomes feasible. This allows a selective reconstruction of the coherent-scatter form factor for a region of interest. The proposed modified 3D filtered back-projection algorithm is a powerful reconstruction technique to be implemented in a CSCT scanning system. This method gives coherent scatter CT the potential of becoming a competitive modality for medical imaging or nondestructive testing. PMID- 14528969 TI - Re-evaluation of the dose to the cyst wall in P-32 radiocolloid treatments of cystic brain tumors using the dose-point-kernel and Monte Carlo methods. AB - Intracavity instillation of beta-emitting colloid pharmaceuticals is a common technique used to treat cystic brain tumors. Most of the dosimetric calculations that have been reported in the literature for this problem are based on empirical formulas derived by Loevinger. Concentration of P-32 radiolabeled solution for the delivery of a prescribed dose (200 Gy to the cyst wall) has been published previously using this formalism in what we refer to as a standard nomogram. The calculations using the Loevinger formulas for calculating the P-32 activity necessary to achieve 200 Gy at the cyst wall is re-evaluated and compared to numerically computed results based on full Monte Carlo simulations (EGSnrc) and the dose-point-kernel (DPK) integration method. For cyst diameters greater than 1 cm, the new calculations agree well with previously published results (the standard nomogram) to within a few percents. However, for cyst diameters of less than 1 cm, it is shown that the standard nomogram results underestimate the therapeutic activity by a factor of approximately 3 for very small diameters (approximately 0.2 cm). New tables based on our calculations are presented and the sources of discrepancies are identified. It is concluded that the new set of data based on our calculations should replace the standard nomogram to administer accurately the target dose to the cyst wall for the smaller diameter cysts (< 1 cm). PMID- 14528970 TI - Dosimetry for radiocolloid therapy of cystic craniopharyngiomas. AB - The dosimetry for radiocolloid therapy of cystic craniopharyngiomas is investigated. Analytical calculations based on the Loevinger and the Berger formulas for electrons and photons, respectively, are compared with Monte Carlo simulations. The role of the material of which the colloid introduced inside the craniopharyngioma is made of as well as that forming the cyst wall is analyzed. It is found that the analytical approaches provide a very good description of the simulated data in the conditions where they can be applied (i.e., in the case of a uniform and infinite homogeneous medium). However, the consideration of the different materials and interfaces produces a strong reduction of the dose delivered to the cyst wall in relation to that predicted by the Loevinger and the Berger formulas. PMID- 14528972 TI - Monte Carlo calculated TG-43 dosimetry parameters for the SeedLink 125Iodine brachytherapy system. AB - The SeedLink brachytherapy system is comprised from an assembly of I-Plant 3500 interstitial brachytherapy seeds and bioresorbable spacers joined together by a 6 mm-long titanium sleeve centered over each seed. This device is designed to maintain specified spacing between seeds during treatment thereby decreasing implant preparation time and reducing radionuclide migration within the prostate and periprostatic region. Reliable clinical treatment and planning applications necessitate accurate dosimetric data for source evaluation, therefore the authors report the results of a Monte Carlo study designed to calculate the AAPM Task Group Report No. 43 dosimetric parameters for the SeedLink brachytherapy source and compare these values against previously published Monte Carlo study results of the I-Plant 3500 brachytherapy seed. For this investigation, a total of 1 x 10(9) source photon histories were processed for each set of in-water and in-air calculations using the MCNP4C2 Monte Carlo radiation transport code (RSICC). Statistically, the radial dose function, g(r), and the dose-rate constant, lambda, were identical to the values calculated previously for the Model 3500 with the dose-rate constant evaluated to be lambda = 1.000+/-0.026 cGyh(-1) U( 1). The titanium sleeve used in SeedLink to bind together Model 3500 seeds and spacers resulted in slightly greater dosimetric anisotropy as exhibited in the anisotropy function, F(r, theta), the anisotropy factor, phi(an) (r), and the anisotropy constant, phi(an), which was calculated to be phi(an) = 0.91 +/- 0.01, or roughly 2% lower than the value calculated previously for the Model 3500. These results indicate that the radiological characteristics of the SeedLink dosimetry system are comparable to those obtained for previously characterized single seeds such as the Implant Sciences Model 3500 I-Plant seed. PMID- 14528971 TI - A quasiexact reconstruction algorithm for helical CT using a 3-Pi acquisition. AB - Recently, an exact reconstruction method for helical CT was published by A. Katsevich. The algorithm is of the filtered backprojection type and is, therefore, computationally efficient. Moreover, during backprojection, only data are used which correspond to an illumination interval of 180 degrees as seen from the object-point. We propose a new reconstruction method, which is applicable to data obtained with a 3-Pi acquisition [IEEE Trans. Med. Imaging 19, 848-863 (2000)]. The method uses the same filter types as the Katsevich algorithm, but the directions and the number of the filter lines are chosen differently. For the derivation of the new algorithm, we analyze the relationship of the Katsevich method and radon inversion. A certain radon plane can intersect with the backprojection interval related to a 3-Pi acquisition either once, three, or five times. In analogy to the definition of quasiexactness introduced by Kudo et al. for a 1-Pi acquisition, we use the term quasiexactness for algorithms on a 3-Pi acquisition, if radon planes with one or three intersections within the backprojection interval are treated correctly. Using the results on the relationship with radon inversion, we can prove that our algorithm is quasiexact in this sense. We use simulation results in order to demonstrate that the algorithm yields excellent image quality. PMID- 14528973 TI - Modeling the instantaneous dose rate dependence of radiation diode detectors. AB - The sensitivity S of Si diodes generally increases with an increase in the instantaneous dose rate r of the radiation beam from clinical linear accelerators. A theoretical model is established to understand the physical origin of this dependence. During a radiation exposure, a portion of the excess minority carriers (electrons or holes) generated in the diode is captured by the R-G (recombination-generation) centers and is recombined with the majority carriers. The captured portion depends on the excess minority-carrier concentration delta p (proportional to r), the R-G center concentration N(t), and the minority-carrier capture cross-sections (sigma(n) for electrons and sigma(p) for holes) by the R-G center. When r increases, the R-G center concentration may not be sufficient to keep the recombination portion constant, which leads to an increase in diode sensitivity because a larger fraction of the charge will be collected. Larger majority-carrier concentration increases the recombination probability of the excess minority carriers and thus decreases the r dependence. The ratio of minority-carrier capture cross-sections, sigma(p)/sigma(n), influences the magnitude of the r dependence and also differentiates the r dependence between n-type and p-type diodes. A number of different circumstances can occur in diodes. When sigma(p) > sigma(n), such as for the dominant R-G center generated by electron radiation, the sensitivity is more dependent on r in an n-type diode than in a p-type diode if all the other device parameters are the same. When sigma(p) < sigma(n), the sensitivity is then more dependent on r in a p-type diode than in an n-type diode. The condition of sigma(p) < sigma(n) can occur when R-G centers with this property are generated by the foundry die process. A diode could have very small r dependence due to large R-G center concentration, generated by heavy platinum doping or radiation accumulated dose. Experimental data are compared with theory. PMID- 14528974 TI - Dose correlation for thoracic motion in radiation therapy of breast cancer. AB - This work investigates the dose correlation for deformed objects due to thoracic motion for radiotherapy treatment of breast cancer. An analytical model has been developed to reconstruct patient anatomy based on the assumption that the body will expand or compress proportionally during respiration. The patient geometry at any phase during a breathing pattern is reconstructed using the CT data taken at the inspiration and expiration phases and the breathing level which can be related to the measured chest wall motion. A correlation between the voxels in the inspiration (or expiration) geometry and the voxels in the reconstructed geometry at any phase of the breathing pattern is established so that the dose can be accumulated during a treatment. The method has been implemented for treatment planning dose calculation by interfacing with a Monte Carlo code. The patient geometry files for different phases of the breathing pattern are generated and the three-dimensional dose data are obtained from the Monte Carlo simulations. The final dose distribution is reconstructed from the dose data at different breathing phases based on patient's breathing pattern associated with chest wall movements. PMID- 14528975 TI - Virtual 3D IVUS vessel model for intravascular brachytherapy planning. I. 3D segmentation, reconstruction, and visualization of coronary artery architecture and orientation. AB - Intravascular brachytherapy (IVB) can significantly reduce the risk of restenosis after interventional treatment of stenotic arteries, if planned and applied correctly. To facilitate computer-based IVB planning, a three-dimensional vessel model has been derived from information on coronary artery segments acquired by intravascular ultrasound (IVUS) and biplane angiography. Part I describes the approach of model construction and presents possibilities of visualization. The vessel model is represented by a voxel volume. Polygonal information about the vessel wall structure is derived by segmentation from a sequence of IVUS images automatically acquired ECG gated during pull back of the IVUS transducer. To detect horizontal, vertical, and radial contours, modified Canny-Edge and Shen Castan filters are applied on Cartesian and polar coordinate representations of the IVUS tomograms as edge detectors. The spatial course of the vessel wall layers is traced in reconstructed longitudinal IVUS scans. By resampling the sequence of IVUS frames the voxel volume is obtained. For this purpose the frames are properly located in space and augmented with additional intermediate frames generated by interpolation. Their spatial location and orientation is derived from biplane X-ray angiography which is performed simultaneously. For resampling, two approaches are proposed: insertion of the vertices of the rectangular goal grid into the cells of a deformed hexahedral mesh derived from the IVUS sequence, and insertion of the vertices of the hexahedral mesh into the cells of the rectangular grid. Finally, the vessel model is visualized by methods of combined volume and polygon rendering. The segmentation process is verified as being in good agreement with results obtained by manual contour tracing with a commercial system. Our approach of construction of the vessel model has been implemented into an interactive software system, 3D IVUS-View, serving as the basis of a future system for intracoronary brachytherapy treatment planning being currently under development (Part II). PMID- 14528976 TI - Monte Carlo calculation of effective attenuation coefficients for various compensator materials. AB - Effective attenuation coefficients for 6, 8, and 15 MV photon beams were derived and studied for various compensator materials for square beams with side lengths of 0.5, 1.0, 2.0, 3.0, and 5.0 cm. Calculations were based on depth dose data in water obtained from EGS4 based DOSXYZ Monte Carlo simulations. Depth dose data were calculated using different compensator materials as attenuators of variable thickness. The absorbed dose varied exponentially as a function of absorber thickness at any depth in water on the beam axis for all materials. The effective attenuation coefficient data were compared with measurements for wax, aluminum and brass with values from the literature. Theoretical narrow beam linear attenuation coefficients were calculated and compared with the Monte Carlo data. The effective attenuation coefficient data for all materials were parametrized as functions of field size and depth in water. The effective attenuation coefficient was also parametrized as a function of atomic number. It was found that the effective attenuation coefficients calculated from the DOSXYZ data using a simple source model correspond to measured data for wax, aluminum and brass and published data for lead. PMID- 14528977 TI - MLC dosimetric characteristics for small field and IMRT applications. AB - The objective of this work was to measure the performance characteristics of a double-focus multileaf collimator (MLC) for intensity modulated radiation therapy (IMRT), specifically the variation in penumbra and leakage for narrow fields as a function of field position over a 20x27 cm space available for segmented MLC IMRT. Measurements were made with 6 MV x rays through a MLC containing 29 leaf pairs (27 pairs of 1 cm width), and EDR2 film at 10 cm depth in solid water at 100 cm SAD. Films were digitized with 0.17 mm resolution and converted to dose. Interleaf and intraleaf transmission were measured along 11 vertical profile locations. Leaf-end transmission was measured along horizontal profiles for each of 9 different leaf abutments, traveling over a 20 cm range. In-plane penumbra measurements were made through a single leaf retracted, for 7 different leaves. Cross-plane penumbra (leaf-end) measurements were made for all 27 leaf pairs, where the 1 cm field width was placed in 11 different off-axis positions (20 cm range). Interleaf leakage (range 1.0%-1.5%), intraleaf transmission (range 0.6% 0.8%), and leaf-end transmission (range 0.8%-2.7%) were consistent for all leaf pairs at a given abutment position. The penumbra for these 1-cm-wide fields was measured to be 0.36 cm+/-0.03 cm for 99% of the measurements. In conclusion, the penumbra and leakage of the double-focus MLC were remarkably consistent for the range of leaf positions studied, producing dosimetric characteristics that are well suited for IMRT segments where opposing leaf pairs are often separated by 10 mm or less. PMID- 14528978 TI - IMRT delivery verification using a spiral phantom. AB - In this paper we report on the testing and verification of a system for IMRT delivery quality assurance that uses a cylindrical solid water phantom with a spiral trajectory for radiographic film placement. This spiral film technique provides more complete dosimetric verification of the entire IMRT treatment than perpendicular film methods, since it samples a three-dimensional dose subspace rather than using measurements at only one or two depths. As an example, the complete analysis of the predicted and measured spiral films is described for an intracranial IMRT treatment case. The results of this analysis are compared to those of a single field perpendicular film technique that is typically used for IMRT QA. The comparison demonstrates that both methods result in a dosimetric error within a clinical tolerance of 5%, however the spiral phantom QA technique provides a more complete dosimetric verification while being less time consuming. To independently verify the dosimetry obtained with the spiral film, the same IMRT treatment was delivered to a similar phantom in which LiF thermoluminescent dosimeters were arranged along the spiral trajectory. The maximum difference between the predicted and measured TLD data for the 1.8 Gy fraction was 0.06 Gy for a TLD located in a high dose gradient region. This further validates the ability of the spiral phantom QA process to accurately verify delivery of an IMRT plan. PMID- 14528979 TI - The emperor's new isodose curves. PMID- 14528980 TI - Barn raising: give kids a smile... every day. PMID- 14528981 TI - Facing a challenge. PMID- 14528982 TI - Caring for the elderly. PMID- 14528983 TI - Fluoride and dermatitis. PMID- 14528984 TI - The whole patient. PMID- 14528985 TI - Oral cancer test. PMID- 14528986 TI - Novel treatments. PMID- 14528987 TI - 'Patch-and-mend'. PMID- 14528988 TI - Bleaching sensitivity. PMID- 14528989 TI - Do you support the ADA's racketeering lawsuit against major insurance carriers? PMID- 14528990 TI - Tooth loss and dietary intake. AB - BACKGROUND: Several studies have reported that impaired dentition status is associated with poor nutritional intake. However, most of these studies are cross sectional and thus are unable to clarify the temporal sequence. METHODS: We assessed the longitudinal relation between tooth loss and changes in consumption of fruits and vegetables and of nutrients important for general health among 31,813 eligible male health professionals. RESULTS: Subjects who lost five or more teeth had a significantly smaller reduction in consumption of dietary cholesterol and vitamin B12, greater reduction in consumption of polyunsaturated fat and smaller increase in consumption of dietary fiber and whole fruit than did subjects who had lost no teeth. Men who had lost teeth also were more likely to stop eating apples, pears and raw carrots. CONCLUSIONS: The results support the temporal association between tooth loss and detrimental changes in dietary intakes, which could contribute to increased risk of developing chronic diseases. PRACTICE IMPLICATIONS: Dietary evaluation and recommendations can be incorporated into dental visits to provide a greater benefit to patients. PMID- 14528991 TI - A pilot study of risk-based prevention in private practice. AB - BACKGROUND: Risk-based prevention is a means of ensuring that patients receive preventive treatment appropriate for their risk of disease. While straightforward, its application in private practice has not been examined. METHODS: Volunteer clinicians in 15 offices participated in a six-month pilot study to test methods for a larger, risk-based prevention demonstration study operated by a dental insurer. Concomitant with oral examinations for patients of this insurer, clinicians identified patients at elevated risk of developing dental caries and periodontitis. For these patients, the reasons for elevated risk (risk indicators), as well as planned preventive treatment in response to that risk, were recorded and transmitted to the insurer via the claim form. RESULTS: The clinicians identified relatively small percentages of patients as being at high risk of developing caries (4 percent) and periodontitis (7 percent), with little variation across the 15 offices. Larger proportions of patients were identified as being at moderate risk of developing caries (29 percent) and periodontitis (30 percent), with more extensive variation across offices. In general, patients classified as being at elevated risk had received more disease-related treatment than patients at low risk before the classification, which provided some validation for the accuracy of risk assessment. CONCLUSIONS: The results of this pilot study suggest that formal, risk-based prevention can be accomplished in dental offices. Clinicians' reported risk assignments and indicators, together with their planned preventive treatments, demonstrate a good understanding of risk-based prevention. CLINICAL IMPLICATIONS: Researchers may need to clarify the criteria used to assess moderate risk of developing dental caries, and clinicians may need to emphasize greater use of fluorides and more frequent recall visits for adults at elevated risk of developing dental caries. PMID- 14528992 TI - Two-year clinical evaluation of a packable resin-based composite. AB - BACKGROUND: Packable resin-based composites were introduced in 1998, but few clinical studies have been conducted to evaluate them. The authors conducted a clinical study to determine the two-year performance of SureFil (Dentsply DeTrey GmbH, Konstanz, Germany) packable posterior resin-based composite in Class I and II restorations. METHODS: An operator (L.S.T.) restored 55 cavities in 36 patients (16 Class I restorations and 39 Class II restorations). After cavity preparation, she etched the enamel with 34 percent phosphoric acid, applied Prime & Bond NT (Dentsply DeTrey GmbH) to dentin and etched enamel for 20 seconds and then cured it for 20 seconds. She restored the cavity using 3- to 5-millimeter increments of SureFil. Independent examiners assessed the restorations after placement and at six months, one year and two years for color matching, marginal discoloration, marginal adaptation, secondary caries, surface texture, anatomical form and postoperative sensitivity, using the Ryge criteria. RESULTS: The authors assessed the changes in the parameters during the two-year period using a software program with Friedman test analysis with a Bonferroni adjustment at significant level of P = .05. At baseline, 31 restorations were graded as Bravo for color match. At the six-month and one-year recall periods (n = 55), 53 restorations remained unchanged. Two restorations from the same patient fell out after one month. After two years (n = 50), there were five Bravos for surface staining and three for marginal adaptation (P < .05). CONCLUSION: After two years of clinical service, SureFil packable resin-based composite had a success rate of 96 percent, and the authors considered it successful in Class I and II restorations. CLINICAL IMPLICATIONS: SureFil packable resin-based composite can be successful in clinical situations with limited-sized cavities and proper application of restorative techniques. PMID- 14528993 TI - Depth of cure and microleakage with high-intensity and ramped resin-based composite curing lights. AB - BACKGROUND: The authors conducted a study to determine whether high-intensity curing lights in high and ramped intensity modes affect microleakage of resin based composite restorations and whether different types of resin-based composites meet American National Standards Institute/American Dental Association Specification no. 27 (1993): 7.7 for depth of cure when polymerized using these lights. METHODS: The authors compared five high-intensity lights, three plasma arc lights and two quartz-tungsten-halogen lights in their regular and ramped intensity modes with a quartz-tungsten-halogen 40-second light. The parameters tested were microleakage one month after bonding and curing depth for different resin-based composite types. The authors measured curing depth using a scratch test. RESULTS: Light curing with Optilux 501 (Kerr/Demetron, Orange, Calif.) for 10 seconds and ADT Power PAC (American Dental Technologies, Corpus Christi, Texas) for 10 seconds resulted in higher microleakage values than light curing with other lights (P < .05). The microhybrid resin-based composite was the only material that met the specification when light cured with all of the lights tested. The flowable resin-based composite did not meet the specification when light cured with all lights tested. Microhybrid resin-based composite had the greatest depth of cure, and flowable resin-based composite had the least depth of cure. CONCLUSIONS: Microhybrid resin-based composite microleakage is affected by some light-curing modes. Different categories of resin-based composites are cured to different depths using high-intensity lights. CLINICAL IMPLICATIONS: Light curing with some high-intensity lights compared with halogen lights may result in higher microleakage values. Use caution when light curing flowable resin-based composite with the high-intensity lights. Place increments less than 2 millimeters in depth when using this material. PMID- 14528994 TI - Adult hemopoietic stem cell transplantation. AB - BACKGROUND: Hemopoietic stem cell transplantation, or HSCT, is an important tool in modern cancer treatment. Refinement of transplantation techniques and supportive care has resulted in increased posttransplantation survival rates. Dental care is a key supportive element in both pretransplantation and posttransplantation care of this patient population. METHODS: The authors provide an overview of HSCT transplantation, emphasizing the oral complications and required supportive dental care. CONCLUSIONS: It is critical that transplantation candidates undergo dental screenings and be treated adequately before transplantation, that their care be closely managed during the transplantation process, and that they be given dental support as soon as their recovery permits. Dentists should consult with the patient's oncologist or primary health provider to identify the appropriate timing and intensity of dental support. CLINICAL IMPLICATIONS: Because of improved transplantation survival rates, more patients may seek supportive outpatient dental care after transplantation, which requires special management considerations. Dental professionals need to be knowledgeable about modern HSCT. PMID- 14528995 TI - Orthodontic treatment with a series of removable appliances. AB - BACKGROUND: In the United States, the demand for straight white teeth has never been more important to patients. Crowded, poorly aligned teeth are not esthetically pleasing and are difficult to keep clean. However, until recently, the process of straightening the teeth typically has involved appliances involving bands, brackets and wires that also can be difficult to clean. The desire for a cosmetic solution to misaligned teeth has led to an increase in the number of patients seeking veneers, crowns and other laboratory-fabricated cosmetic restorations. Some clinicians are not aware that there are other ways to align teeth without either significant enamel reduction or conspicuous fixed orthodontic appliances. DESCRIPTION OF TECHNIQUE: An alternative method of treatment, involving a series of clear removable appliances, circumvents this shortcoming and enables tooth alignment while avoiding deterioration in the cosmetics of the smile during treatment. This article describes a method of treatment, Invisalign (Align Technology, Santa Clara, Calif.), that clinicians can use in conjunction with 3-D computer models to accomplish the esthetic and oral hygiene objectives during and after treatment. CASE DESCRIPTIONS: The authors present three case reports, all involving a chief concern of crowding. The first case involved treatment of both arches by interproximal reduction, alignment of teeth and leveling the curve of Spee. The second case involved treatment of both arches by proclination, expansion and minor interproximal reduction. The final case shows relief of lower-arch crowding via lower-incisor extraction. CLINICAL IMPLICATIONS: In addition to satisfying the patient's chief concern of desiring straight teeth, this method of treatment satisfies additional objectives of esthetic treatment and esthetic results with significant oral hygiene benefits. Clinicians can address a patient's chief concern effectively without requiring an inventory of appliances (such as bands, brackets, wires and instruments). Furthermore, the use of 3-D computer models can give clinicians additional information to make more complete decisions regarding treatment. PMID- 14528996 TI - Manual versus powered toothbrushes: the Cochrane review. AB - BACKGROUND: In a report released in January 2003, The Cochrane Collaboration Oral Health Group provided a systematic review of the effectiveness of powered versus manual toothbrushes, which generated much interest in the popular press. METHODS: The Cochrane researchers developed and implemented search strategies for the Cochrane Oral Health Group's Trial Register, the Cochrane Central Register of Controlled Trials, MEDLINE and the Cumulative Index to Nursing and Allied Health Literature. They contacted manufacturers for additional information. Trials were selected if they met the following criteria: compared power versus manual toothbrushes, had a randomized design, drew participants from a general population without disabilities, provided data regarding plaque and gingivitis, and were at least 28 days in length. Reviewers evaluated only studies published in 2001 or earlier. Six reviewers from the Cochrane study independently extracted information in duplicate. Indexes for plaque and gingivitis were expressed as standardized values for data distillation. Data distillation was accomplished using a meta-analysis, with a mean difference between powered and manual toothbrushes serving as the measure of effectiveness. RESULTS: The searches identified 354 trials, only 29 of which met the inclusion criteria. These trials involved about 2,500 subjects and provided data for meta-analysis. The results indicated that only the rotating oscillating toothbrush consistently provided a statistically significant, although modest, clinical benefit over manual toothbrushes in reducing plaque and gingivitis. CONCLUSIONS AND CLINICAL IMPLICATIONS: Some powered toothbrushes with a rotation-oscillation action achieve a significant, but modest, reduction in plaque and gingivitis compared with manual toothbrushes. PMID- 14528997 TI - Physician group agrees to settle price-fixing charges. Underscores hazards of collectively negotiating fees. PMID- 14528998 TI - The confusing array of tooth-colored crowns. AB - The venerable PFM crown or fixed prosthesis still dominates the tooth-colored restoration market. However, use of PFMs is declining slightly, as the many new all-ceramic and resin-based composite crowns and fixed-prosthesis products flood the market. Several situations may indicate the use of materials other than PFM. They include patients requiring a high level of esthetic acceptability, patients with proven or perceived allergies to the metals used in dentistry and bruxing or clenching patients with metal allergies or desire to eliminate metal from their mouths. PFM restorations have been proven during 40 years of successful use. They provide acceptable esthetics for most situations, minimal fracture during service, proven ability to serve in multiple-unit situations and excellent fit, and the profession has detailed knowledge of these restorations' advantages, disadvantages and physical characteristics. PFM restorations have only a few well known negative characteristics. Be cautious as you elect to move from the reliability and positive history of PFM to relatively unknown types of restorations. PMID- 14528999 TI - Nitrile gloves. PMID- 14529000 TI - Setting your practice goals. AB - Goal setting is a powerful business technique. Far more than just a motivational concept, it is an outcome of strategic planning. Doctors who set goals are usually more clearly focused on where they want the practice to go and when they want to get there. PMID- 14529001 TI - A legal perspective on antibiotic prophylaxis. PMID- 14529002 TI - For the dental patient. The facts about bottled water. PMID- 14529003 TI - Physiology of spinal anaesthesia and practical suggestions for successful spinal anaesthesia. AB - There are numerous physiological effects of spinal anaesthesia. This chapter focuses on the physiological effects that are of clinical relevance to the anaesthesiologist, and provides suggestions for successful management of this simple and popular technique. The mechanisms and clinical significance of spinal anaesthesia-induced hypotension, bradycardia and cardiac arrest are reviewed. The increasing popularity of ambulatory spinal anaesthesia requires knowledge that long-acting local anaesthetics, such as bupivacaine, impair the ability to void far longer than short-acting local anaesthetics, such as lidocaine. The importance of thermoregulation during spinal anaesthesia, and the clinical consequences of spinal-anaesthesia-induced hypothermia are reviewed. Effects of spinal anaesthesia on ventilatory mechanics are also highlighted. Lastly, the sedative and minimum-alveolar-concentration-sparing effects of spinal anaesthesia are discussed to reinforce the need for the judicious use of sedation in the perioperative setting. PMID- 14529004 TI - Local anaesthetics and additives for spinal anaesthesia--characteristics and factors influencing the spread and duration of the block. AB - Different characteristics of patients and local anaesthetic formulations will influence the spread of spinal anaesthesia. The predictability of the spread of spinal anaesthesia can be improved by altering both baricity of the solution, and the position of the patient during the intrathecal local anaesthetic injection. The role of adrenaline and clonidine in prolonging the block and associated side effects is discussed. The role of opioids added to local anaesthetic solutions is discussed from a cost/benefit point of view. PMID- 14529005 TI - Spinal anaesthesia for spinal surgery. AB - Spinal anaesthesia for spinal surgery is becoming increasingly more popular because this anaesthetic technique allows the patient to self-position and avoid neurological injury that may occur with prone positioning under general anaesthesia. Spinal anaesthesia reduces intraoperative surgical blood loss, improves perioperative haemodynamic stability and reduces pain in the immediate postoperative period. This leads to a reduced need for analgesics and a reduction in the incidence of nausea and vomiting in the postoperative setting. Spinal anaesthesia for lumbar spine surgery also decreases the incidence of lower extremity thrombo-embolic complications and does not increase the occurrence of problems with micturition. These benefits increase the patient's satisfaction, and they expedite discharge of the patient from the hospital. Combination anaesthetic techniques, using both subarachnoid and epidural dosing schemes, may be beneficial for improving postoperative pain control and add further to the benefit of spinal anaesthesia for lumbar spine surgical procedures. PMID- 14529006 TI - Spinal anaesthesia for outpatient surgery. AB - Spinal anaesthesia in the outpatient is characterized by rapid onset and offset, easy administration, minimal expense, and minimal side effects or complications. Spinal anaesthesia offers advantages for outpatient lower extremity, perineal, and many abdominal and gynaecological procedures. Development of small-gauge, pencil-point needles are responsible for the success of outpatient spinal anaesthesia with acceptable rates (0-2%) of postdural puncture headache (PDPH). Compared with peripheral nerve blocks, spinal anaesthesia has a more predictable offset. There are many possible choices of local anaesthetics for outpatient spinal anaesthesia. These include lidocaine, prilocaine, mepivacaine and small doses of bupivacaine. Meperidine has local anaesthetic properties in addition to its opiate properties. It has been used as the sole intrathecal agent for spinal anaesthesia but has no real advantages over lidocaine. Mepivacaine and lidocaine have each been associated with transient neurological symptoms (TNS) following intrathecal administration. This has stimulated development of alternative agents, including combinations of local anaesthetics and opioids. Lidocaine remains the most useful agent for outpatient spinal anaesthesia. For longer procedures, mepivacaine is an excellent spinal anaesthetic agent. Attention to technique, reduction of dose and addition of fentanyl to lidocaine result in effective spinal anaesthesia with rapid recovery and a low incidence of significant side effects or complications. PMID- 14529007 TI - The combined spinal--epidural technique. AB - In recent years, the use of regional anaesthesia techniques for surgery, obstetrics and post operative pain management have increased in popularity. The combined spinal-epidural (CSE) technique has attained widespread popularity for patients undergoing major surgery below the umbilicus who may require prolonged and effective postoperative analgesia. The CSE technique is now well established in several institutions. This chapter includes the clinical experience, advantages and potential problems, and discusses future perspectives of the CSE technique. PMID- 14529008 TI - Spinal anaesthesia in paediatrics. AB - The use of spinal anaesthesia in children has been primarily limited to situations in which general anaesthesia was considered to pose an excessive risk. The ex-premature infant and the neurologically impaired child account for the majority of spinal anaesthetics used today. Spinal anaesthesia, compared with general anaesthesia, in the ex-premature infant undergoing inguinal hernia repair has decreased postoperative respiratory complications (e.g. apnoeic events, prolonged mechanical ventilation). Hyperbaric tetracaine and bupivacaine solutions are the local anaesthetics of choice. Haemodynamic stability is well preserved in neonates having spinal anaesthesia. Advances in spinal needle design have decreased the incidence of postdural puncture headache (PDPH). Catastrophic events have occurred with neuraxial techniques. Care must be taken in evaluating the relative risks of anaesthetic approaches in infants and children. PMID- 14529009 TI - Spinal anaesthesia for obstetrics. AB - For a long time, epidural anaesthesia has been considered the method of choice for Caesarean delivery. The increased incidence of hypotension by the rapid onset of sympathetic blockade under spinal anaesthesia has been associated with a decline in uteroplacental blood flow and significant fetal acidosis, which may compromise neonatal well-being. Nevertheless, a decrease in fetal pH has not been shown to reduce neonatal Apgar or neurobehavioural assessment scores. Maternal blood pressure can be preserved with little side effects with low doses of vasopressors. On the other hand, spinal anaesthesia conveys significant advantages over epidural anaesthesia such as the simplicity of its use and the speed of onset, which allows neuraxial anaesthesia in urgent Caesarean sections and thus reduces the necessity for general anaesthesia. The small doses of local anaesthetics required to perform spinal anaesthesia reduce the risks of systemic toxicity to zero. Spinal anaesthesia is now considered the method of choice for urgent Caesarean section. The use of intrathecal opioids has profoundly changed the quality of spinal anaesthesia, with improved analgesia, a reduction in local anaesthetic requirements and shorter duration of motor blockade. Preliminary studies indicate that spinal anaesthesia may be safely performed in patients with severe pre-eclampsia, in whom spinal anaesthesia was previously considered contraindicated. PMID- 14529010 TI - Continuous spinal anaesthesia: what's new and what's not. AB - Continuous spinal anaesthesia combines the advantages of single-dose spinal anaesthesia, rapid onset and a high degree of success, with those of a continuous technique. The introduction of micro-catheters invigorated interest in the technique and allowed its expansion to additional populations and surgical procedures. However, multiple cases of cauda equina syndrome associated with micro-catheters and (primarily) hyperbaric lidocaine solution led to withdrawal of micro-catheters from the US market, casting doubt over the safety of continuous spinal anaesthesia as a whole. A decade after these events it is possible to look back at the experience with continuous spinal anaesthesia for operative anaesthesia and postoperative analgesia and to compare it with the available alternatives. From this perspective, continuous spinal anaesthesia remains a useful and safe technique. Future research should focus on the comparison of continuous spinal anaesthesia with the combined spinal/epidural technique and the use of newer spinal agents. PMID- 14529011 TI - Intrathecal neurolytic blocks for the relief of cancer pain. AB - Intrathecal neurolytic blocks for the treatment of chronic pain were first described by Dogliotti in 1931. Since then, many authors have described the intrathecal injection of various neurolytic substances for the treatment of oncologic pain. In recent years, alcohol and phenol have been the substances most commonly used for this purpose. There are no controlled studies, so the literature consists of observations, reports and book chapters reflecting the opinions of experienced clinicians. This chapter describes the indications, contraindications, potential complications, and expected benefits of intrathecal injection of alcohol and phenol in the treatment of cancer pain. Four cases of cancer patients whose intractable pain was treated by the authors using intrathecal neurolysis are presented. Pertinent literature is reviewed. In this account, the emphasis is on proper selection of patients and techniques. PMID- 14529012 TI - Intrathecal morphine for chronic benign pain. AB - In the treatment of chronic benign pain, the administration of an intrathecal opioid produces a potent analgesia without interfering with the motor and sensory functions of the lower extremities. An intrathecal opioid should be considered only when pain control with conventional oral and systemic administration is inadequate or is associated with unmanageable side effects. A trial period and a psychological evaluation are mandatory prior to implantation of a permanent device. About 40% of the patients need surgical revision for various complications. Hormonal changes may influence sexual behaviour. Catheter granulomas can form with high concentrations of morphine. Adjuvant drugs such as bupivacaine, clonidine and ketamine might be necessary to deal with the development of tolerance to morphine. The sophistication of available technology for intrathecal infusion today far exceeds our knowledge of the potential neurological effects of this treatment modality. PMID- 14529014 TI - Post-dural puncture headache: pathophysiology, prevention and treatment. AB - Post-dural puncture headache (PDPHA) has been a vexing problem for patients undergoing dural puncture for spinal anaesthesia, as a complication of epidural anaesthesia, and after diagnostic lumbar puncture since Bier reported the first case in 1898. This Chapter discusses the pathophysiology of low-pressure headache resulting from leakage of cerebrospinal fluid (CSF) from the subarachnoid to the epidural spaces. Clinical and laboratory research over the last 30 years has shown that use of small-gauge needles, particularly of the pencil-point design, is associated with a lower risk of PDPHA than traditional cutting point needle tips (Quincke-point needles). A careful history can rule out other causes of headache. A positional component of headache is the sine qua non of PDPHA. In high-risk patients (e.g. age < 50 years, post-partum, large-gauge-needle puncture), patients should be offered early (within 24-48 h of dural puncture) epidural blood patch. The optimum volume of blood has been shown to be 12-20 ml for adult patients. Complications of autologous epidural blood patch are rare. PMID- 14529013 TI - Spinal anaesthesia and the use of anticoagulants. AB - This chapter addresses the increasing incidence of spinal haematoma after central neuraxis anaesthesia in patients receiving drugs that affect coagulation. Administration of low-molecular-weight heparins in the perioperative period is highlighted because these drugs remain the 'gold standard' for prophylaxis against deep-vein thrombosis. The performance of spinal anaesthesia in patients already receiving antiplatelet drugs is discussed--as well as special warnings in such a setting. In addition, issues such as those concerning the administration of unfractionated heparin, anti-vitamin K drugs or new antiplatelet and antithrombotic medications are addressed. Finally, specific recommendations regarding each class of drug are defined in order to avoid the occurrence of a rare but catastrophic event such as spinal haematoma. PMID- 14529015 TI - Neurotoxicity of intrathecal local anaesthetics and transient neurological symptoms. AB - Local anaesthetics have been placed in the intrathecal space for approximately 100 years. Currently used intrathecal local anaesthetics appear to be relatively benign on the basis of the low incidence of permanent neurological deficits. In large retrospective surveys of 4000-10 000 patients, the incidence of persistent neurological sequelae after subarachnoid anaesthesia varies between 0.01 and 0.7%. Since its introduction in 1948, hyperbaric 5% lidocaine has been used for millions of spinal anaesthetics. The predictable onset and limited duration of action have made lidocaine one of the most popular spinal anaesthetics currently available. Concern about the use of spinal lidocaine began in 1991 with published reports of cauda equina syndrome after continuous spinal anaesthesia. In 1993, Schneider published a case report of four patients undergoing spinal anaesthesia who postoperatively experienced aching and pain in the buttocks and lower extremities. This chapter reviews the neurotoxicity of spinal local anaesthetics, as well as the incidence, possible aetiology, and treatment of transient neurological symptoms after lidocaine spinal anaesthesia. PMID- 14529016 TI - Ion/molecule reactions performed in a miniature cylindrical ion trap mass spectrometer. AB - A recently constructed miniature mass spectrometer, based on a cylindrical ion trap (CIT) mass analyzer, is used to perform ion/molecule reactions in order to improve selectivity for in situ analysis of explosives and chemical warfare agent simulants. Six different reactions are explored, including several of the Eberlin reaction type (M. N. Eberlin and R. G. Cooks, Org. Mass Spectrom., 1993, 28, 679 687) as well as novel gas-phase Meerwein reactions. The reactions include (1) Eberlin transacetalization of the benzoyl, 2,2-dimethyloximinium, and 2,2 dimethylthiooximinium cations with 2,2-dimethyl-1,3-dioxolane to form 2-phenyl 1,3-dioxolanylium cations, 2,2-dimethylamine-1,3-dioxolanylium cations and the 2,2-dimethylamin-1,3-oxathiolanylium cations, respectively; (2) Eberlin reaction of the phosphonium ion CH3P(O)OCH3+, formed from the chemical warfare agent simulant dimethyl methylphosphonate (DMMP), with 1,4-dioxane to yield the 1,3,2 dioxaphospholanium ion, a new characteristic reaction for phosphate ester detection; (3) the novel Meerwein reaction of the ion CH3P(O)OCH3+ with propylene sulfide forming 1,3,2-oxathionylphospholanium ion; (4) the Meerwein reaction of the benzoyl cation with propylene oxide and propylene sulfide to form 4-methyl-2 phenyl-1,3-dioxolane and its thio analog, respectively; (5) ketalization of the benzoyl cation with ethylene glycol to form the 2-phenyl-1,3-dioxolanylium cation; (6) addition/NO2 elimination involving benzonitrile radical cation in reaction with nitrobenzene to form an arylated nitrile, a diagnostic reaction for explosives detection and (7) simple methanol addition to the C7H7+ ion, formed by NO2 loss from the molecular ion of p-nitrotoluene to form an intact adduct. Evidence is provided that these reactions occur to give the products described and their potential analytical utility is discussed. PMID- 14529017 TI - Solid phase micro-extraction in a miniature ion trap mass spectrometer. AB - Fiber introduction mass spectrometry (FIMS), a variation of solid-phase microextraction (SPME) and membrane introduction mass spectrometry (MIMS), is employed with a miniature mass spectrometer. The inlet system, constructed of commercially available vacuum parts, allows the direct introduction of the SPME needle vacuum chamber into the mass spectrometer. Thermal desorption of the analyte from the poly(dimethylsiloxane) (PDMS) coated fiber was achieved with a built in nichrome heater, followed by electron ionization of the analytes internal to the cylindrical ion trap (CIT). The system has been tested with several volatile organic compounds (VOC) in air and to analyze the headspace over aqueous solutions, with limits of detection in the low ppb range. The signal rise (10-90%) and fall (90-10%) times for the system ranged from 0.1 to 1 s (rise) and 1.2 to 6 s (fall) using heated desorption. In addition, this method has been applied to quantitation of toluene in benzene, toluene, xylene (BTX) mixtures in water and gasoline. This simple and rapid analysis method, coupled to a portable mass spectrometer, has been shown to provide a robust, simple, rapid, reproducible, accurate and sensitive (low ppb range) fieldable approach to the effective in situ analysis of VOC in various matrices. PMID- 14529018 TI - Micro sequential injection: automated insulin derivatization and separation using a lab-on-valve capillary electrophoresis system. AB - Automated sampling and fluorogenic derivatization of islet proteins (insulin, proinsulin, c-peptide) are separated and analyzed by a novel lab-on-valve capillary electrophoresis (LOV-CE) system. This fully integrated device is based on a micro sequential injection instrument that uses a lab-on-valve manifold to integrate capillary electrophoresis. The lab-on-valve manifold is used to perform all microfluidic tasks such as sampling, fluorogenic labeling, and CE capillary rejuvenation providing a very reliable system for reproducible CE separations. Fluorescence detection was coupled to an epiluminescence fluorescence microscope using a customized capillary positioning plate. This customized plate incorporated two fused-silica fiber optic probes that allow for simultaneous absorbance and fluorescence detection, extending the utility of this device. Derivatization conditions with respect to the sequence of addition, timing, injection position, and volumes were optimized through iterative series of experiments that are executed automatically by software control. Reproducibility in fluorogenic labeling was tested with repetitive injections of 3.45 mM insulin, yielding 1.3% RSD for peak area, 0.5% RSD for electromigration time, and 2.8% RSD for peak height. Fluorescence detection demonstrated a linear dynamic range of 3.43 to 6.87 microM for insulin (r2 = 0.99999), 0.39 to 1.96 pM for proinsulin (r2 = 0.99195) and 260 to 781 nM for c-peptide (r2 = 0.99983). By including hydrodynamic flushing immediately after the detection of the last analyte, the sampling frequency for islet protein analysis was increased. Finally, an in vitro insulin assay using rat pancreatic islet excretions was tested using this lab-on valve capillary electrophoresis system. PMID- 14529019 TI - Determination of the ascorbate content of photographic developer solutions using miniaturised isotachophoresis on a planar chip. AB - The use of miniaturised isotachophoresis, performed on a planar poly(methyl methacrylate) device with integrated platinum conductivity electrodes, for the analysis of the ascorbate content of photographic developer solutions has been investigated. An electrolyte system has been developed which enabled the analysis to be made without interference from any of the other components in the developer solution, a number of which were present at significantly higher concentrations than that of the ascorbate ions. Using this system, the ascorbate content of the developer solutions could be analysed in under 6 min. The limit of detection for ascorbate using miniaturised isotachophoresis was calculated to be 0.011 mmol dm( 3). PMID- 14529020 TI - PDMS-based microfluidic device with multi-height structures fabricated by single step photolithography using printed circuit board as masters. AB - We have developed a method for fabricating microfluidic devices with multi-height structures using single step photolithography. The whole fabrication process is executed by conventional printed circuit board (PCB) technology without the need of having access to clean room facilities. Specifically designed "windows" and "rims" architectures were printed on films that were used as photomasks. Different levels of protruding features on the PCB master were produced by exposing a photomask followed by chemical wet etching. Poly(dimethylsiloxane) (PDMS) was then moulded against the positive relief master to generate microfluidic structures. In this report, we described the fabrication of a microfluidic device featured with a multi-height "sandbag" structure for particle entrapment and peripheral microchannels. Controlled immobilization of biological cells and immunocytochemcial staining assays were performed to demonstrate the applicability of the microfluidic device for cellular analysis. The integrity of the microdevice remained stable under applied pressure, indicating the robustness of the elastic PDMS structures for analytical operation. The simple microfabrication process requires only low-cost materials and minimal specialized equipment and can reproducibly produce mask lines of about 20 microm in width, which is sufficient for most microfluidic applications. PMID- 14529021 TI - Water-soluble cyclodextrin polymers for enhanced relative recovery of hydrophobic analytes during microdialysis sampling. AB - Microdialysis relative recovery (RR) enhancement using different water-soluble, epichlorohydrin-based cyclodextrin polymers (CD-EPS) was studied in vitro for different analytes, amitryptiline, carbamazepine, hydroquinone, ibuprofen, and 4 nitrophenol. When compared to the native CDs (alpha, beta, and gamma) on a per mole basis, the CD-EPS enhanced microdialysis RR was either statistically greater or the same. beta-CD-EPS was more highly retained than native beta-CD by a 20 000 Da molecular weight cutoff (MWCO) polycarbonate membrane, but showed no statistical difference for loss across a 100 000 Da MWCO polyethersulfone membrane (PES). When the same weight percent of beta-CD or beta-CD-EPS was included in the microdialysis perfusion fluid, the beta-CD-EPS produced a higher microdialysis RR than native beta-CD for all analytes across the PES membrane. However, enhancements for the PC membrane were statistically insignificant when beta-CD and beta-CD-EPS were compared on a per mole basis. These results suggest that CD-EPS may be used as effective enhancement agents during microdialysis sampling and for some membranes provide the additional advantage of being retained more than native CDs. PMID- 14529022 TI - A universal temperature controlled membrane interface for the analysis of volatile and semi-volatile organic compounds. AB - A universal temperature controlled membrane interface (TCMI) has been constructed for hollow-fibre membranes. The membrane temperature is controllable in the range -70 to 250 degrees C using an electric heater and a flow of cooled nitrogen or helium gas. Volatile and semi-volatile organic compounds may be detected either by continuous diffusion across the membrane or by in-membrane pre-concentration followed by thermal desorption into the detector. The TCMI interface is demonstrated in combination with mass spectrometry and GC-MS, for the determination of VOCs and SVOCs in aqueous and air samples and for the on-line monitoring of a bioreactor. PMID- 14529023 TI - Bead injection ELISA for the determination of antibodies implicated in type 1 diabetes mellitus. AB - This work introduces a novel analytical method for the detection and study of GAD65 autoantibodies, which have been implicated in the onset of type 1 diabetes. There is a clinical need for a rapid and automated assay for GAD65 autoantibodies. Therefore, this method was designed to exploit the advantages of bead injection (BI) analysis for enzyme-linked immunosorbent assays (ELISA). BI ELISA is a microscale technique that uses enzyme labeled secondary antibodies to detect the capture of target antibodies on immobilized antigen in the flow cell of the lab-on-valve (LOV) manifold. A detection limit of 20 ng mL(-1) of GAD65 monoclonal antibody 144 compares favorably with the sensitivity and precision of a standard ELISA currently employed to detect GAD65 autoantibodies. Compared to the standard ELISA protocol, BI ELISA offers a significantly reduced assay time and complete automation of solution handling and detection. PMID- 14529024 TI - Determination of erythrocyte deformability and its correlation to cellular ATP release using microbore tubing with diameters that approximate resistance vessels in vivo. AB - A novel method is described for measuring the deformability of red blood cells (RBCs) in tubing whose diameters approximate forces encountered in vivo. Here, RBCs from rabbits are loaded into a 50 cm section of 75 microm id microbore tubing and connected to a syringe pump. This section of tubing is then connected to a 15 cm section of 25 microm id tubing. As buffer is pumped through the flow system, the RBCs are evacuated from both sections of tubing. However, the inability of the RBCs to move freely through the 25 mirom id section of tubing results in a buildup of cells at the inlet of this portion of tubing. The continued force output by the syringe pump results in a deformation of the RBCs until all of the cells are eventually evacuated from the flow system. It was found that a measurement of the time required to reach half of the maximum pressure (1/2 P(max)) may be used as an indicator of the RBC deformability. For a given sample, a simple buffer results in less time to reach 1/2 P(max) (6.9 +/- 0.2 s) than deformable RBCs (21.6 +/- 0.8 s). To verify that the increased amount of time to reach 1/2 P(max) is indeed due to the RBCs, various hematocrits of an RBC sample were investigated and, as expected, it was found that a 12% RBC hematocrit had a higher 1/2 P(max) value (26.0 s +/- 2.2 s) when compared to a 7% hematocrit (19.1 +/- 0.3 s). In addition, RBCs chemically stiffened with glutaraldehyde were shown to be 25% less deformable than normal RBCs. Finally, a study was performed to examine the relationship between RBC deformability and ATP release and it was found that ATP release increased as a function of RBC deformability. This method greatly simplifies deformability measurements, employing only a syringe pump and microbore tubing, and may lead to a more complete understanding of the physiological significance of erythrocyte deformability. PMID- 14529025 TI - Effects of DNA mismatches on binding affinity and kinetics of polymerase-DNA complexes as revealed by surface plasmon resonance biosensor. AB - In this study, surface plasmon resonance (SPR) biosensor techniques were used to obtain quantitative information on the kinetics of the DNA and polymerase I (Klenow fragment) interaction. DNA duplexes containing different base compositions at the binding site were immobilized on the SPR sensor surface via biotin-streptavidin chemistry and the subsequent binding of the polymerase was measured in real time. Various kinetic models were tested and a translocation model was shown to provide the best fit for the binding and dissociation profiles. The results revealed that the enzyme binds to DNA at both the polymerase and the exonuclease domains with different association and dissociation rates as well as affinity constants, depending on the presence of mismatches near the primer 3'-end. Introduction of unpaired bases increases the DNA binding affinity towards the exonuclease domain and promotes the translocation of DNA from the polymerase site to the exonuclease site. The results also demonstrated that SPR biosensors may be used as a sensitive technique for studying molecular recognition events such as single-base discrimination involved in protein-DNA interaction. PMID- 14529026 TI - Hypersonic evanescent waves generated with a planar spiral coil. AB - A planar spiral coil has been used to induce hypersonic evanescent waves in a quartz substrate with the unique ability to focus the acoustic wave down onto the chemical recognition layer. These special sensing conditions were achieved by investigating the application of a radio frequency current to a coaxial waveguide and spiral coil, so that wideband repeating electrical resonance conditions could be established over the MHz to GHz frequency range. At a selected operating frequency of 1.09 GHz, the evanescent wave depth of a quartz crystal hypersonic resonance is reduced to 17 nm, minimising unwanted coupling to the bulk fluid. Verification of the validity of the hypersonic resonance was carried out by characterising the system electrically and acoustically: Impedance calculations of the combined coil and coaxial waveguide demonstrated an excellent fit to the measured data, although above 400 MHz a transition zone was identified where unwanted impedance is parasitic of the coil influence efficiency, so the signal to-noise ratio is reduced from 3000 to 300. Acoustic quartz crystal resonances at intervals of precisely 13.2138 MHz spacing, from the 6.6 MHz ultrasonic range and onto the desired hypersonic range above 1 GHz, were incrementally detected. Q factor measurements demonstrated that reductions in energy lost from the resonator to the fluid interface were consistent with the anticipated shrinkage of the evanescent wave with increasing operating frequency. Amplitude and frequency reduction in contact with a glucose solution was demonstrated at 1.09 GHz. The complex physical conditions arising at the solid-liquid interface under hypersonic entrainment are discussed with respect to acceleration induced slippage, rupture, longitudinal and shear radiation and multiphase relaxation affects. PMID- 14529027 TI - Rapid method for the preparation of a robust optical pH sensor. AB - A simple and rapid method for the preparation of a fluorescence-based optical pH sensor is described. The sensor is based on excitation ratiometric detection of a methacryloyl-modified analog of the well-known fluorescent pH indicator dye, 8 hydroxy-1,3,6-pyrene trisulfonic acid (HPTS). The modified dye, 6-methacryloyl-8 hydroxy-1,3-pyrene disulfonic acid (MA-HPDS), is similar in structure and function to HPTS. However, unlike HPTS, the presence of the methacrylate moiety allows MA-HPDS to participate as a comonomer in a free radical polymerization reaction. Covalent immobilization by this method is simple, as the preparation of the modified dye is accomplished in a single reaction step and its subsequent reaction with a comonomer results in simultaneous immobilization and purification. Sensors were prepared by copolymerization of the MA-HPDS with poly(ethylene glycol) diacrylate. Minimal leaching of the immobilized dye was observed from the HPDS-PEG matrix. The copolymer of HPDS and PEG-DA is fully autoclavable. The sensor is useful over the pH range of 6-9, with excellent reproducibility. Ionic strength effects on the apparent pKa of the immobilized dye are small and predictable. PMID- 14529028 TI - Selective silver ion transfer voltammetry at the polarised liquid/liquid interface. AB - Transfer of silver ions across the water/1,2-dichloroethane interface was studied by cyclic voltammetry (CV). In the absence of added neutral ionophore, Ag+ transferred across the interface when the organic phase contained either tetraphenylborate or tetrakis(4-chloro)phenylborate anions, but this transfer was not possible in the presence of organic phase hexafluorophosphate or perchlorate anions. The ion transfer processes observed were independent of the nature of the organic phase cation. The CV in the presence of tetraphenylborate exhibited a shape consistent with an ion transfer followed by chemical reaction; the rate constant for the following chemical reaction was 0.016 s(-1). In the presence of tetrakis(4-chloro)phenylborate, a return peak equivalent in magnitude to the forward peak was observed, indicative of a simple ion transfer reaction uncomplicated by accompanying chemical reactions. The selectivity of the transfer was assessed with respect to other metal cations: no transfers for copper, cadmium, lead, bismuth, cobalt, nickel, palladium or zinc were observed. The selectivity of the transfer suggests this can form the basis of a selective voltammetric methodology for the determination of silver ions. PMID- 14529030 TI - Elimination of interference information by a new hybrid algorithm for quantitative calibration of near infrared spectra. AB - A new hybrid algorithm is proposed to eliminate the interference information for multivariate calibration of near-infrared (NIR) spectra that includes noise, background and systemic spectral variation irrelevant to concentration. The method consists of two parts: approximate derivative based on continuous wavelet transform (CWT) and orthogonal signal correction (OSC). After the approximate derivative calculated by CWT, OSC was performed. It was successfully applied to real complex NIR spectral data to eliminate the interference information. Correction for the interference of NIR spectra resulted in a substantial improvement in the predicted precision, and a more concise calibration model was obtained. The proposed procedure also compared favourably with several pretreatment methods, and the new method appears to provide a high-performance pretreatment tool for multivariate calibration of NIR spectra. In addition, the strategy proposed here can be applied to various other spectral data for quantitative purposes as well. PMID- 14529029 TI - Classification of commercial apple beverages using a minimum set of mid-IR wavenumbers selected by Procrustes rotation. AB - When infrared spectral data are used in classification and/or multivariate regression methods there can be problems related to both chemical understanding and computation speed due to the large number of wavenumbers in each spectrum. Here, it is shown that the Procrustes rotation technique can be used to select a minimum set of spectral variables (wavenumbers) to perform classification and regression. Procrustes rotation was coupled to several multivariate methods as PLS, SIMCA and potential curves (a maximum likelihood classification method). The practical problem of implementing a screening methodology for classifying apple juice-based beverages according to their contents of "pure" apple juice was addressed using attenuated total reflectance, mid-IR spectroscopy. It is found that two of the original wavenumbers are almost as good predictors as all the 176 initial ones. PMID- 14529031 TI - Determination of total sulfur in diesel fuel employing NIR spectroscopy and multivariate calibration. AB - A method for sulfur determination in diesel fuel employing near infrared spectroscopy, variable selection and multivariate calibration is described. The performances of principal component regression (PCR) and partial least square (PLS) chemometric methods were compared with those shown by multiple linear regression (MLR), performed after variable selection based on the genetic algorithm (GA) or the successive projection algorithm (SPA). Ninety seven diesel samples were divided into three sets (41 for calibration, 30 for internal validation and 26 for external validation), each of them covering the full range of sulfur concentrations (from 0.07 to 0.33% w/w). Transflectance measurements were performed from 850 to 1800 nm. Although principal component analysis identified the presence of three groups, PLS, PCR and MLR provided models whose predicting capabilities were independent of the diesel type. Calibration with PLS and PCR employing all the 454 wavelengths provided root mean square errors of prediction (RMSEP) of 0.036% and 0.043% for the validation set, respectively. The use of GA and SPA for variable selection provided calibration models based on 19 and 9 wavelengths, with a RMSEP of 0.031% (PLS-GA), 0.022% (MLR-SPA) and 0.034% (MLR-GA). As the ASTM 4294 method allows a reproducibility of 0.05%, it can be concluded that a method based on NIR spectroscopy and multivariate calibration can be employed for the determination of sulfur in diesel fuels. Furthermore, the selection of variables can provide more robust calibration models and SPA provided more parsimonious models than GA. PMID- 14529032 TI - Advance directives: burden or benefit? PMID- 14529033 TI - Interdisciplinary teamwork in palliative care and hospice settings. PMID- 14529034 TI - Reducing obstacles to rural hospice care. PMID- 14529036 TI - End-of-life care education in United States pharmacy schools. AB - Hospice and palliative care have undergone dramatic changes in the past 30 years. Educational initiatives and certification programs for physicians (American Board of Hospice and Palliative Medicine) and nurses (National Board for Certification of Hospice and Palliative Nurses) have further delineated this area of practice as distinct from geriatrics, neurology, anesthesiology, or oncology. As other professions assess their own practices of hospice and end-of-life (EOL) care education in their respective schools and colleges, the pharmacy profession must also ensure that its future graduates are prepared to adequately participate in this type of care. This was a descriptive study in which all accredited schools and colleges of pharmacy in the US were queried regarding their level of curricular commitment to EOL care. Eighty-three questionnaires were mailed, and 60 schools responded (72 percent). Four primary informational items regarding EOL and palliative care education were targeted, including availability of didactic teaching, specialization of pharmacy faculty, availability and type of clerkships, and method of instruction. Sixty-two percent of respondents indicated EOL care education was provided didactically (3.89 +/- 1.91 lecture hours per year). Fifty-eight percent of respondents indicated that EOL care experiential clerkships were available (4.97 +/- 1.25 weeks in duration). These data indicate that over half of US pharmacy students receive some exposure to EOL care education. PMID- 14529035 TI - Guidelines for breakthrough pain dosing. PMID- 14529037 TI - The business of palliative medicine--Part 3: The development of a palliative medicine program in an academic medical center. AB - Palliative medicine is the total continuing care of patients with cancer. Most resources for cancer care focus on curative attempts while often ignoring the symptoms created by the disease and its treatment. Attempts at curative treatment of the malignancy must be coupled with pain and symptom relief psychosocial and spiritual care, and support for the patient and family extending from the time of diagnosis through the bereavement period. To accomplish this important goal, we must establish comprehensive palliative medicine programs in cancer centers throughout the world. These programs must include education, research, and patient care and must work through an interdisciplinary team. The Cleveland Clinic Foundation palliative medicine program (PMP) is composed of a primary inpatient service, consult service, outpatient clinic, hospice homecare, and cancer homecare services. In this article, we describe the structure and development of the program and suggest future avenues for growth. PMID- 14529038 TI - Advance care planning among residents in long-term care. AB - This study was conducted to determine whether two types of advance directives exist for individuals residing in long-term care facilities. Findings were based on data from the Medical Expenditure Panel Study-Nursing Home Component (MEPS NHC), a survey using a two-stage stratified probability sample of nursing homes and residents to produce valid national estimates of the nursing home population in the United States. The two types of advance directives included basic, i.e., living will or do-not-resuscitate (DNR) order, and progressive (do-not hospitalize order or orders restricting feeding, medication, or other treatment). Approximately 59 percent of long-term care residents had a basic advance directive, 9 percent have a progressive directive, and 60 percent have some type of directive. Logistic regression results indicate that the factors associated with the likelihood of each type of directive differ considerably, and only two variables (African American ethnicity and less time in the facility) were associated with a reduced likelihood of having either type of directive. Our results indicate that the two proposed types of advance directives are distinct with regard to the variables predicting each. PMID- 14529039 TI - Satisfaction of families using end-of-life care: current successes and challenges in the hospice industry. AB - This study examined the satisfaction of family members with the end-of-life care their loved ones received. Data were collected from 1,839 individuals receiving care from 17 different care agencies nationwide. Although family satisfaction with hospice care was generally quite high. situational factors played a role. The timing of the referral was critical, with families rating services lower almost across the board when the referral to hospice was deemed "too late." Additionally, families expressed greater satisfaction when the patient's care was overseen by the hospice director, rather than a personal physician. Each of these findings has important implications for physicians, patients, and families as they begin to plan for end-of-life care. PMID- 14529040 TI - An analysis of provider attitudes toward end-of-life decision-making. AB - The explosion of technological and biomedical interventions over the past half century appears to have enhanced the medical profession's ability to prolong life at a faster pace than society's ability to develop comprehensive bioethical laws governing end-of-life decisions. This study was conducted to determine if there was a correlation between educational and occupational backgrounds and participants' perceptions of end-of-life care. Study participants came from various areas and levels of healthcare at a large federal healthcare facility and included nurses, doctors, pharmacists, dietitians, administrators, and others. Participants completed a questionnaire providing both sociodemographic information and perceptions of end-of-life care. Respondents were classified by type of education or background (technical, professional, or medical) and by level of involvement with patient care (direct or indirect). While the results showed a general consensus about the importance of respecting patients' end-of life wishes, there were differences among respondents in regard to specific issues. PMID- 14529041 TI - Toward evidence-based prescribing at end of life: a comparative review of temazepam and zolpidem for the treatment of insomnia. AB - A comparative review of temazepam and zolpidem use in managing insomnia in the hospice patient was undertaken to determine whether treatment with temazepam is a more cost-effective approach for this patient population. A MEDLINE search was conducted to identify pertinent literature, including clinical trials and reviews that involved temazepam or zolpidem. Published data was used as background information and provided in the discussion. This retrospective analysis, conducted from June 2002 through November 2002, focused on the prescribing patterns of temazepam and zolpidem in our hospice practice setting. We examined the reasons for discontinuation of each agent, along with the frequency of therapeutic change from temazepam to zolpidem. The top 10 ICD-9 codes associated with each treatment modality were investigated to determine any prescribing patterns. A total of 4,752 participants were prescribed either temazepam or zolpidem during this six-month period. Of the 4,065 patients prescribed temazepam 9.9 percent had the agent discontinued, whereas, 13.0 percent of those taking zolpidem (n = 687) terminated therapy. Reasons for discontinuation included change in dose, incomplete efficacy, change in patient status, adverse drug reaction, cultural/social issues and "other." Analyses of prescribing patterns and the reasons for termination of each drug therapy were completed and compared with results found in the primary literature. Due to the limited financial resources available for hospice care, our goal is to provide the most clinically appropriate and cost-effective agents for hospice patients. With the lack of data pertaining to the hospice patient, physicians often are faced with challenges in deciding the most appropriate therapy. They may prefer one agent over another based on current medical opinion rather than sound clinical evidence. After review of the primary literature and the prescribing patterns in our setting, there is currently no evidence in our patient population to support that zolpidem is superior to benzodiazepines for the treatment of insomnia. PMID- 14529042 TI - Treatable complications of cancer patients referred to an in-patient hospice. AB - This paper illustrates the importance of accurate diagnoses and treatments of complications in terminally ill cancer patients. The paper reports on five hospice in-patients who completely recovered from life-threatening complications; three of them had been incorrectly labeled as "imminently dying" by the referring physicians. The paper concludes that it would be beneficial for patients to receive examinations and a trial of medical treatment in their continuing treatment settings. PMID- 14529043 TI - Questions, answers, and the bridge of language. PMID- 14529044 TI - Ultrastructural correlates of synapse withdrawal at axotomized neuromuscular junctions in mutant and transgenic mice expressing the Wld gene. AB - We carried out an ultrastructural analysis of axotomized synaptic terminals in Wld(s) and Ube4b/Nmnat (Wld) transgenic mice, in which severed distal axons are protected from Wallerian degeneration. Previous studies have suggested that axotomy in juvenile (< 2 months) Wld mice induced a progressive nerve terminal withdrawal from motor endplates. In this study we confirm that axotomy-induced terminal withdrawal occurs in the absence of all major ultrastructural characteristics of Wallerian degeneration. Pre- and post-synaptic membranes showed no signs of disruption or fragmentation, synaptic vesicle densities remained at pre-axotomy levels, the numbers of synaptic vesicles clustered towards presynaptic active zones did not diminish, and mitochondria retained their membranes and cristae. However, motor nerve terminal ultrastructure was measurably different following axotomy in Wld transgenic 4836 line mice, which strongly express Wld protein: axotomized presynaptic terminals were retained, but many were significantly depleted of synaptic vesicles. These findings suggest that the Wld gene interacts with the mechanisms regulating transmitter release and vesicle recycling. PMID- 14529045 TI - Age-associated plasticity in the intrinsic innervation of the ovine rumen. AB - The rumen of adult sheep functions as a large fermentation chamber. In the newborn suckling ruminant, the rumen is bypassed and milk enters the abomasum directly. It was the aim of our study to investigate whether the transmitter content of intrinsic nerves changes with the developmental stage. The neurochemical code of myenteric neurons in the rumen from suckling lambs, fattened lambs and adult sheep was determined by using quadruple immunohistochemistry against choline-acetyltransferase (ChAT), nitric oxide synthase (NOS), substance P (SP) and vasoactive intestinal peptide (VIP). Three neurochemically distinct subpopulations were identified within the rumen. They expressed the code ChAT/-, ChAT/SP and NOS/VIP. The number of ChAT/SP neurons did not change during development. It was 62% in the newborn lamb and remained stable in fattened lambs (63%) and adult sheep (63%). By contrast, the number of ChAT/- neurons decreased significantly from 20% in suckling lambs to 11% and 7% in fattened lambs and adult sheep, respectively. Simultaneously, the proportion of NOS/VIP neurons increased from 16% in suckling lambs to 29% in adult sheep. The increase in the proportion of NOS/VIP immunoreactive neurons indicates an adaptation to large volumes of ingesta at the beginning of roughage intake and rumination. We conclude that the age-associated changes in neurochemical code of myenteric neurons in the forestomach are related to the adaption of the rumen to different functional properties during development. PMID- 14529046 TI - Zebrin II compartmentation of the cerebellum in a basal insectivore, the Madagascan hedgehog tenrec Echinops telfairi. AB - The mammalian cerebellum is histologically uniform. However, underlying the simple laminar architecture is a complex arrangement of parasagittal stripes and transverse zones that can be revealed by the expression of zebrin II/aldolase C. The cerebellar cortex of rodents, for example, is organized into four transverse zones: anterior, central, posterior and nodular. Within the anterior and posterior zones, parasagittal stripes of Purkinje cells expressing zebrin II alternate with those that do not. Zonal boundaries appear to be independent of cerebellar lobulation. To explore this model further, and to broaden our understanding of the evolution of cerebellar patterning, zebrin II expression has been studied in the cerebellum of the Madagascan hedgehog tenrec (Echinops telfairi), a basal insectivore with a lissiform cerebellum with only five lobules. Zebrin II expression in the tenrec reveals an array of four transverse zones as in rodents, two with homogeneous zebrin II expression, two further subdivided into stripes, that closely resembles the expression pattern described in other mammals. We conclude that a zone-and-stripe organization may be a common feature of the mammalian cerebellar vermis and hemispheres, and that zonal boundaries and cerebellar lobules and fissures form independently. PMID- 14529048 TI - Pectoral fin development in the Baikalian viviparous golomyankas (Comephoridae; Cottoidei), with a remark on eggs and embryos of Comephorus baicalensis (Pallas). AB - Measurements of pectoral fin (PF) growth in relation to standard body length (SL) for Comephorus dybowskii (SL: 14-126 mm) and C. baicalensis (SL: 22-173 mm) were made and compared with corresponding data obtained for 13 other Baikalian cottoid species (sculpins). The allometry for the PF/SL relationships (Y = a + bX) is clearly biphasic. Expressed in percentages, these values for the larval period of C. dybowskii [Yp = -28.886 + (2.419X)] are much higher than for C. baicalensis [Yp = -11.904 + (1.233X)]. The postnatal period of development of the golomyanka up to 30-35 mm SL is considered to be the larval stage. In the postlarval period, growth of pectoral fin and the body in C. dybowskii are directly proportional [Yp' = 48.92 + (-0.009X)] whereas in C. baicalensis they rise slightly [Yp' = 30.966 + (0.0505X)]. Absence of the air bladder characterizes all Baikalian cottoid species, including the golomyankas. Various species of the golomyanka differ in the ways of obtaining neutral buoyancy. It is supposed that large pectoral fins guarantee behavioural buoyancy in small species such as C. dybowskii, whereas in C. baicalensis, obtaining much larger body size but lower PFp/SL index, the buoyancy is gained mainly due to the accumulation of large amount of lipids. This paper also documents the ovary and advanced embryos of C. baicalensis. PMID- 14529047 TI - Segmentation in staged human embryos: the occipitocervical region revisited. AB - The first seven somites, the rhombomeres, and the pharyngeal arches were reassessed in 145 serially sectioned human embryos of stages 9-23, 22 of which were controlled by precise graphic reconstructions. Segmentation begins in the neuromeres, somites and aortic arches at stage 9. The following new observations are presented. (1) The first somite in the human, unlike that of the chick, is neither reduced in size nor different in structure, and it possesses sclerotome, somitocoel and dermatomyotome. (2) Somites 1-4, unlike those of the chick, are related to rhombomere 8 (rather than 7 and 8) and are caudal to pharyngeal arch 4 (rather than in line with 3 and 4). (3) Occipital segment 4 resembles a developing vertebra more than do segments 1-3. (4) The development of the basioccipital resembles that of the first two cervical vertebrae in that medial and lateral components arise in a manner that differs from that in the rest of the vertebral column. (5) The two groups of somites, occipital 1-4 and cervical 5 7, each form a median skeletal mass. (6) An 'S-shaped head/trunk interface', described for the chick and unjustifiably for the mouse, was not found because it is not compatible with the topographical development of the otic primordium and somite 1, between which neural crest migrates without hindrance in mammals. (7) Occipital segmentation and related features are documented by photomicrographs and graphic interpretations for the first time in the human. It is confirmed that the first somite, unlike that of the chick, is separated from the otic primordium by a distance, although the otic anlage undergoes a relative shift caudally. The important, although frequently neglected, distinction between lateral and medial components is emphasized. Laterally, sclerotomes 3 and 4 delineate the hypoglossal foramen, 4 gives rise to the exoccipital and participates in the occipital condyle, 5 forms the posterior arch of the atlas and 6 provides the neural arch of the axis, which is greater in height than the arches of the other cervical vertebrae. Medially, the perinotochord and migrated sclerotomic cells give rise to the basioccipital as well as to the vertebral centra, including the tripartite column of the axis. Registration between (1) the somites and (2) the occipital and cervical medial segments becomes interrupted by the special development of the axis, the three components of which come to occupy the height of only 2 1/2 segments. PMID- 14529049 TI - In vivo time-course of the angiogenic response induced by multiple myeloma plasma cells in the chick embryo chorioallantoic membrane. AB - In this study, we set out to make a fine characterization of the angiogenic response induced by plasma cells obtained from patients with active-multiple myeloma (MM), in comparison with cells obtained from patients with non-active MM and benign lesions such as monoclonal gammopathy of undetermined significance (MGUS), in the chick embryo chorioallantoic membrane (CAM) assay. To achieve this we investigated the time-course of the angiogenic response induced by gelatin sponges soaked in the cell suspensions and implanted on the CAM surface from day 8 to day 12 of incubation by evaluating the number of vessels, of the vessel bifurcation and the intervascular distance at 24, 48, 72 and 96 h after the implants. The results show that plasma cell suspensions obtained from patients with active MM induce a vasoproliferative response that was significantly higher than that induced by cell suspensions obtained from patients with non-active MM or with MGUS, which is also a function of the day of implantation. In fact, implants made from day 8 to day 10 induce a strong angiogenic response, whereas those made from day 11 to day 12 do not. This finding might depend on the fact that CAM endothelium exhibits an intrinsically high mitotic rate until day 10. Thereafter, the endothelial mitotic index declines rapidly, and consequently cell suspensions implanted on the CAM of successively older embryos are not able to induce a vasoproliferative response in parallel with the reduced rates of growth of the CAM's endothelial cells. PMID- 14529051 TI - Estimation of nuclear volume as an indicator of maturation of glial precursor cells in the developing rat spinal cord: a stereological approach. AB - Studies on nuclear volume have shown that it is an indication of the state of differentiation of cells. This study provides evidence indicating increasing nuclear volume during cell maturation. Using unbiased stereological techniques, nuclear volume of both proliferating and non-proliferating glial cells was analysed in the developing spinal cord. Proliferating glial precursor cells were identified using a 5-bromo-2'-deoxyuridine (BrdU) incorporation assay. The nuclear volume of BrdU-labelled cells and unlabelled cells was determined in both periventricular regions and the white matter of the cord at different embryonic ages. In the periventricular region BrdU-labelled nuclei were smaller than unlabelled nuclei at all ages examined. These labelled cells represent dividing undifferentiated progenitors. The unlabelled neighbouring cells with larger nuclei represent a more differentiated population. In the white matter BrdU labelled nuclei were of similar volume to the unlabelled nuclei. Both of these groups represent glial precursor cells that have migrated from deeper regions and are at similar stages of differentiation, perhaps with different proliferative potential. These findings indicate that the nuclear volume of early glial cells increases as these cells migrate and differentiate. PMID- 14529050 TI - Immunohistochemistry of the canine vomeronasal organ. AB - The canine's olfactory acuity is legendary, but neither its main olfactory system nor its vomeronasal system has been described in much detail. We used immunohistochemistry on paraffin-embedded sections of male and female adult dog vomeronasal organ (VNO) to characterize the expression of proteins known to be expressed in the VNO of several other mammals. Basal cell bodies were more apparent in each section than in rodent VNO and expressed immunoreactivity to anticytokeratin and antiepidermal growth factor receptor antibodies. The thin layer of neurone cell bodies in the sensory epithelium and axon fascicles in the lamina propria expressed immunoreactivity to neurone cell adhesion molecule, neurone-specific beta tubulin and protein gene product 9.5. Some neurones expressed growth-associated protein 43 (GAP43): and a number of those also expressed neurone-specific beta tubulin-immunoreactivity. Some axon fascicles were double labelled for those two proteins. The G-protein alpha subunits Gi and Go, involved in the signal transduction pathway, showed immunoreactivity in the sensory cell layer. Our results demonstrate that the canine vomeronasal organ contains a population of cells that expresses several neuronal markers. Furthermore, GAP43 immunoreactivity suggests that the sensory epithelium is neurogenic in adult dogs. PMID- 14529052 TI - Prejudice and perspective. PMID- 14529053 TI - Autoimmune hepatitis caused by Brucella: causal or a mere association? PMID- 14529054 TI - Comparison of once-daily mometasone furoate versus once-daily budesonide in patients with moderate persistent asthma. AB - We conducted this study to compare the efficacy and safety of once-daily mometasone furoate (MF) administered by dry powder inhaler (DPI) with once-daily budesonide (BUD)-DPI and placebo in patients with moderate persistent asthma previously using twice-daily inhaled corticosteroids. A total of 262 patients (> or = 12 years of age) with moderate persistent asthma were randomised to once daily morning treatment with MF-DPI 440 microg (metered dose), BUD-DPI 400 microg (metered dose), or placebo in an eight-week, multicentre, placebo-controlled, double-blind, double-dummy study. The primary efficacy variable was percent change in FEV1 from baseline to endpoint (last evaluable visit). At endpoint, the percent change in FEV1 was significantly greater (p < 0.01) following treatment with MF-DPI 440 microg (8.9%) than with both BUD-DPI 400 microg (2.1%) and placebo (-3.9%). Secondary efficacy variables, including morning and evening peak expiratory flow rates, albuterol use, percentage of asthma symptom-free days, and physician-evaluated response to therapy were also significantly improved at endpoint in the MF-DPI group compared with both the placebo and BUD-DPI groups (p < 0.05). Both active treatments were well tolerated. In conclusion, once-daily treatment in the morning with MF-DPI 440 microg significantly improved pulmonary function and asthma control compared with morning administration of BUD-DPI 400 microg and placebo. PMID- 14529056 TI - The value of coagulation profiles in epistaxis management. AB - A prospective pilot study was conducted to investigate the role and relevance of clotting profiles in the management of epistaxis patients. Forty-eight per cent of the study patients had abnormal clotting profiles. Half of the abnormal clotting profiles could have been predicted from a routine history, but half could not. The coagulation screens on this group were repeated and 5% of the initial study group were found to have a persistent abnormality. The authors suggest that although coagulation screens will not predict the potential management problems associated with the epistaxis patient, they remain an important part of the clinical investigation. An epistaxis may be the only indication of an underlying coagulation disorder. The diagnosis of such a disorder following an epistaxis may lead to the treatment of an otherwise unknown condition and prevention of more serious sequelae. PMID- 14529055 TI - Patients with migraine prefer zolmitriptan orally disintegrating tablet to sumatriptan conventional oral tablet. AB - The Zolmitriptan Evaluation versus Sumatriptan Trial (ZEST) assessed patient preference for 2.5 mg zolmitriptan orally disintegrating tablet (ODT) or 50 mg sumatriptan conventional tablet in 218 patients with significant migraine disability. Significantly more patients preferred zolmitriptan ODT to sumatriptan conventional tablet (60.1% vs 39.9%; p = 0.0130). In terms of efficacy, significantly more patients considered zolmitriptan ODT to be an effective migraine treatment than sumatriptan conventional tablet (77% vs 63%; p = 0.0063). When asked about specific formulation attributes, significantly more patients selected zolmitriptan ODT as the least disruptive therapy (83.6% vs 16.4%), the easiest to take (85.5% vs 14.5%), the most convenient to take (86.1% vs 13.9%), and the one which enabled them to maintain an active lifestyle (65.5% vs 34.5%), compared with the sumatriptan conventional tablet (all comparisons p < 0.001). Zolmitriptan ODT is a convenient and beneficial alternative to conventional tablets and is preferred to sumatriptan conventional tablets by migraineurs. PMID- 14529057 TI - Reusable versus disposable instruments in tonsillectomy: a comparative study of outcomes. AB - Tonsillectomy, traditionally performed using reusable instruments (RIs), has recently been performed using disposable instruments (DIs), to prevent the theoretical risk of spreading new variant Creutzfeldt-Jakob disease. Following this, there have been concerns of increased risk of complications, especially haemorrhage. We audited our hospital's complication rates of 100 tonsillectomies performed using RI and 100 using DIs. We analysed the overall complication rates and the postoperative haemorrhage rates. and compared them with the broader groups of RIs versus DIs. We also compared patients who did not have diathermy with those who did have diathermy. The overall complication rate was 6% with RIs and 33% with DIs and the haemorrhage rate was 1% and 12%, respectively. With DIs, these rates were significantly higher in the diathermy subgroup. We believe this is the first study to look at the complications due to DIs. PMID- 14529058 TI - Sympathetic blockade and amitriptyline in the treatment of reflex sympathetic dystrophy. AB - We prospectively investigated the outcome of a combination therapy of oral amitriptyline and sympathetic ganglion blockade on 10 patients suffering from reflex sympathetic dystrophy of the upper extremity for at least three months. The efficacy of the treatment was evaluated by clinical examination, pain ratings on the visual analogue scale (VAS) and grip strength measurements using the Jamar dynamometer. The results were statistically analysed with Wilcoxon signed-ranks test for comparison of the before and after treatment grip strength measurements and with paired t-test for comparison of the mean of initial and consecutive pain ratings on the VAS. Values of p < 0.01 were considered to be statistically significant. Combination therapy proved beneficial in this particular patient population, which, as far as previously documented studies are concerned, would otherwise respond less favourably to a treatment consisting solely of sympathetic blockade. PMID- 14529059 TI - Superior mesenteric arterial embolism: a retrospective study of local thrombolytic treatment with urokinase in West China. AB - Embolism of the superior mesenteric artery (SMA) is a rare disorder. To explore the selective criteria of local thrombolysis, evaluate its efficacy and discuss the therapeutic protocol of SMA embolism, a retrospective case control study was undertaken. Sixteen cases were divided into two groups: nine cases (group A) from May 1995 to April 1999 were given traditional treatment, while seven cases (group B) from November 1999 to June 2002 received comprehensive therapy including local thrombolysis. The local thrombolytic treatment was performed without procedure related complications and the embolus was successfully lysed in four patients. The time between admission and diagnosis (or treatment) was shorter in group B than in group A (2.3 +/- 1.2 vs 12.3 +/- 9.2 hr, p = 0.013). Gangrenous bowel segments in group A were much longer than in group B (159.4 +/- 87.7 vs 45.7 +/- 61.6 cm, p = 0.009). However, mortality between the two groups was the same (p = 0.282, OR = 0.32, 95% CI = 0.039, 2.618) perhaps because of the small sample size. Angiography could shorten the duration from the onset of SMA embolism to therapy and certainly lowers the risk of mortality. Local fibrinolytic infusion may be an effective alternative to embolectomy in patients with SMA embolism but without intestinal infarction. PMID- 14529060 TI - Fever of unknown origin: a report from China of 208 cases. AB - Fever of unknown origin (FUO) is a clinical dilemma in western countries and in China. To investigate the causes and prognosis of FUO, 208 patients with FUO admitted to a large university hospital in China were investigated. The final diagnoses established in 158 cases (75.96%) were: infectious disease in 66 cases (31.73%), collagen vascular disease in 46 patients (22.11%), neoplasm in 35 cases (16.83%), and other disease in 11 patients (5.29%). In 66 cases with infectious disease, tuberculosis, septicaemia and typhoid fever were the principal causes. SLE and adult Still's disease were the most important causes among collagen vascular disease. Lymphoma and malignant histiocytosis were mostly associated with FUO among neoplasms. In 50 cases (24.04%), the cause of fever was not found. On discharge from hospital, fever had subsided in 133 cases (63.94%), and had persisted in 63 cases (30.29%); 12 patients (5.87%) died. In China, infectious disease, collagen vascular disease and neoplasm are the main causes of FUO. While most patients recover, there are some differences in the distribution of causes between the West and China, and there are relatively more deaths than in previous reports. PMID- 14529061 TI - Clinical trials of sildenafil citrate (Viagra) demonstrate no increase in risk of myocardial infarction and cardiovascular death compared with placebo. AB - We pooled data regarding myocardial infarction (MI) and cardiovascular death from more than 120 clinical trials of sildenafil citrate (Viagra) conducted from 1993 to 2001. During placebo-controlled trials, the rate of MI or cardiovascular death was 0.91 (95% CI: 0.52-1.48) per 100 person-years (PY) of follow-up among sildenafil-treated patients compared with 0.84 (95% CI: 0.39-1.60) per 100 PY of follow-up among placebo-treated patients. The relative risk of MI or cardiovascular death was 1.08 (95% CI: 0.45-2.77) for sildenafil compared with placebo (p = 0.88). During open-label studies, the rate of MI or cardiovascular death was 0.56 (95% CI: 0.44-0.72) per 100 PY of follow-up. This analysis showed that the rates of MI and cardiovascular death were low and comparable between men treated with sildenafil and those treated with placebo. The use of sildenafil was not associated with an increase in the risk of MI or cardiovascular death. PMID- 14529062 TI - The potential value of erectile dysfunction inquiry and management. AB - Erectile dysfunction (ED) is a serious condition that becomes more common as men age. Many older men, however, report satisfactory erectile capacity and enjoy satisfying sexual relationships. Physicians have been slow to discuss ED with patients even in the presence of multiple risk factors. New information provides strong reasons for ED inquiry and management in the primary care physician's office. The presence of ED can reveal as yet undiscovered neurovascular and psychological disorders including diabetes, hypertension, dyslipidaemia, depression and anxiety as well as early neuromuscular disorders. By inquiring about ED, physicians can better decrease iatrogenic sexual dysfunction caused by certain commonly used medications. The successful management of ED, made much easier by the development of phosphodiesterase type 5 inhibitors, has additional potential benefits including improvement of quality of life for both the patient and his partner; decreasing the symptoms of depression in depressed men who also have ED; improving relationships, a significant factor related to good health; and enhancing overall patient health. Other potential values for the physician include a better clinician-patient and increased physician work satisfaction. Primary care physicians need to recognise the value of ED inquiry and management and integrate these activities into practice. PMID- 14529063 TI - Peginterferon alfa-2a (40KD) (Pegasys) for the treatment of patients with chronic hepatitis C. AB - Compared with conventional interferon alfa, peginterferon alfa-2a (40KD) has improved pharmacokinetics, provides sustained therapeutic plasma levels, and can be administered once weekly. In randomised, multinational trials, peginterferon alfa-2a (40KD) 180 microg once weekly was significantly more effective than three times weekly interferon alfa-2a in patients with chronic hepatitis C, including patients with cirrhosis. Peginterferon alfa-2a (40KD) and ribavirin 1000/1200 mg/day for 48 weeks produced significantly higher sustained responses than three times weekly interferon alfa-2b and ribavirin 1000/1200 mg/day in patients with chronic hepatitis C including those with HCV genotype 1, genotypes 2/3 and those with high or low viral loads at baseline. The drug is well tolerated when given alone or in combination with ribavirin. Health-related quality of life was significantly less impaired during treatment with peginterferon alfa-2a (40KD) than interferon alfa-2a in randomised trials. Peginterferon alfa-2a (40KD) is widely approved for use in patients with chronic hepatitis C. PMID- 14529064 TI - Levetiracetam: a new therapeutic option for refractory epilepsy. AB - Levetiracetam (LEV) is the most recently licensed antiepileptic drug (AED) for adjunctive therapy of partial seizures. Its mechanism of action is uncertain but it exhibits a unique profile of anticonvulsant activity in models of chronic epilepsy. Three randomised, double-blind, placebo-controlled trials enrolling 904 patients with refractory partial epilepsy have demonstrated the efficacy of LEV as adjunctive therapy, with a responder rate (> or = 50% reduction in seizure frequency) of 28-41%. Long-term efficacy studies suggest retention rates of 60% after one year, with 13% of patients seizure-free for six months of the study and 8% seizure-free for one year. Adverse effects of LEV, including somnolence, lethargy and dizziness, were generally mild and the frequency of incidents was not significantly different between the active treatment and placebo groups in clinical trials. LEV has no clinically significant pharmacokinetic interactions (PKI) with other AEDs, or with commonly prescribed medications. Preliminary data suggest that LEV has efficacy in primary generalised epilepsy and further randomised trials are under way. The combination of potent antiepileptic properties with a relatively mild adverse effect profile makes LEV an attractive adjunctive therapy for partial seizures. PMID- 14529066 TI - The impact of implementing the Ottawa ankle rules on ankle radiography requests in A&E. AB - This audit was set up to quantify the effect of implementing the Ottawa ankle rules in a district general hospital that relies on both medical and nursing radiography requests. Data were collected prospectively on 207 patients who presented with an acute ankle injury between August 2001 and February 2002. The department's activity was recorded before and after a period of teaching on the Ottawa ankle rules. Before teaching, 71% of patients with an acute ankle injury were sent for radiography; teaching reduced this figure to 56% (p < 0.05). Auditing the activity of our department enabled us to observe a significant decrease in the number of patients sent for ankle radiography following acute ankle injury. This correlates well with research in other settings. The difficulties of rationalising radiology investigations are discussed. PMID- 14529065 TI - Kounis syndrome secondary to allergic reaction following shellfish ingestion. AB - Two cases of allergic angina and allergic myocardial infarction (Kounis syndrome) secondary to shellfish ingestion are described. The patients had pre-existing quiescent coronary artery disease (type II variant of the syndrome) and the allergic reaction following eating shellfish seemed to have triggered the development of an acute myocardial infarction. The clinical implications are also discussed. PMID- 14529068 TI - Hyperkalaemia in a patient with Graves' disease. AB - A case of hyporeninaemic hypoaldosteronism in a young girl with treated Graves' disease presenting as hyperkalaemia is described. The possible autoimmune link of these two conditions is discussed. PMID- 14529067 TI - Carotid artery invasion by squamous cell carcinoma of the upper aerodigestive tract: the predictive value of CT imaging. AB - Carotid artery invasion by squamous cell carcinoma of the upper aerodigestive tract is a poor prognostic indicator and a relative indicator of tumour inoperability. A number of imaging modalities are available for detecting such invasion, but, because of its availability, computerised tomography (CT) is the usual modality. We have analysed the CT scans of 196 patients referred to the department with an upper aerodigestive tract squamous cell carcinoma. Comparing the scans with the operative findings indicates that CT scans significantly overestimate carotid artery invasion. PMID- 14529069 TI - Gluteal compartment syndrome misdiagnosed as deep vein thrombosis. AB - Gluteal compartment syndrome usually occurs secondary to prolonged immobility and recumbency following alcohol and drug abuse. The resultant pain and swelling in the lower limb may be misdiagnosed as deep vein thrombosis. Once compartment syndrome is diagnosed, urgent decompression fasciotomy is required. The outcome is poor if the diagnosis is missed or delayed. We report such a case. PMID- 14529070 TI - An unusual cause of carpal tunnel syndrome. AB - Tuberculosis is an uncommon cause of carpal tunnel syndrome. We report a patient with systemic lupus erythematosus who developed hand numbness. Ultrasonic, operative and microbiological findings supported the diagnosis of median nerve compression secondary to a tuberculous abscess. The symptoms resolved with surgical excision and antituberculous chemotherapy. PMID- 14529071 TI - Intrathyroidal thyroglossal duct cyst presenting as a thyroid nodule. AB - A 50-year-old woman presented with a lateral neck swelling, clinically indistinguishable from solitary thyroid nodule. A right hemithyroidectomy was performed and microscopy revealed an intrathyroidal thyroglossal cyst. Intrathyroidal thyroglossal cyst should be considered in the differential diagnosis of a thyroid nodule. PMID- 14529072 TI - Autoimmune hepatitis triggered by Brucella infection or doxycycline or both. AB - Autoimmune hepatitis is a disorder of unknown aetiology in which progressive destruction of the hepatic parenchyma occurs, often progressing to cirrhosis. Hepatitis A, Ebstein-Barr virus and measles virus have been identified as triggers for autoimmune hepatitis in susceptible individuals. There are also reports about herbal medicine and minocycline. A case with autoimmune hepatitis triggered by Brucella infection or doxycycline, or both, is presented. An 11-year old female patient treated with six weeks of doxycycline and three weeks of streptomycine for brucellosis presented with histologically proven autoimmune hepatitis (AH) and responded to corticosteroid treatment. Since neither brucellosis nor doxcycyline as triggering factors for AH have been described so far, these two entities are discussed and the literature reviewed. PMID- 14529073 TI - Treatment of toxic epidermal necrolysis with intravenous immunoglobulin. AB - Toxic epidermal necrolysis (TEN) is a rare severe reaction of the skin resulting in full thickness damage to the epidermis. The condition has significant morbidity as a result of dehydration, protein loss, thermoregulatory difficulties, and renal, lung, liver and heart failure. The mortality rate approaches 30%, most commonly from bacterial sepsis. Management of this condition is cessation of the suspected causative agent and supportive care on a burns or intensive care unit. There have been recent reports of treatment using intravenous immunoglobulin (IVIG) therapy, though its efficacy is yet to be established. It has been proposed that IVIG inhibits the Fas-FasL mediated apoptosis of keratinocytes affected by TEN. We describe a case of extensive drug induced TEN in a 33-year-old woman who showed rapid improvement with IVIG therapy at a dose of 0.75 g/kg/day given for four consecutive days. PMID- 14529074 TI - Plummer-Vinson syndrome: a report of three cases. AB - Plummer-Vinson syndrome is characterised by dysphagia, anaemia, glossitis and oesophageal web. We report our findings in three patients with membranes in the upper oesophagus. All patients underwent endoscopic dilatation and iron replacement therapy, with good results. We review the literature of this syndrome. PMID- 14529075 TI - A cross-cultural comparison of British and Japanese lay theories of schizophrenia. AB - Nearly 300 matched British and Japanese participants completed an equivalent three-part questionnaire in their native language. The questionnaire covered general beliefs or conceptions about schizophrenia, causal explanations for the aetiology of schizophrenia, as well as the role of hospitals in particular, and society in general, in helping schizophrenics recover from their illness. It was predicted that the Japanese, who have more taboos about mental illness than the British, would see schizophrenics as more difficult, dangerous and "morally insane". Factor analysis of each of the three parts of the questionnaire yielded a clearly interpretable structure. The British were more concerned with the rights of schizophrenics and believed them to be less dangerous and abnormal than did the Japanese. The Japanese favoured sociological (stress) explanations more than the British for the cause of schizophrenia. Whereas the Japanese saw micro- and macro-society change as the best way to help schizophrenics, the British stressed individual care and consideration as more relevant. PMID- 14529077 TI - The ethnic density effect: results from a national community survey of England and Wales. AB - Using data from a community survey of 5196 ethnic minority and 2867 white respondents, together with data on local group concentration from the 1991 Census, the hypothesis was tested that ethnic group concentration is associated with lower levels of reported psychiatric symptoms. The hypothesis was broadly confirmed, both for within- and between-group differences. However, the effect was found to be modest in size and in one group, the Pakistani sample, was reversed. The findings are inconsistent with an explanation based on selection or drift. Linguistic factors contributed to, but did not explain the effects. Evidence on victimisation and mutual support suggests that social causation, in the form of reduced exposure to direct prejudice and increased social support, is a likely cause of the effect. PMID- 14529076 TI - Leaving the asylum: a psychodynamic observation study of a move from a long stay psychiatric ward to the community. AB - Regular observations were made of a group of residents and staff on a long stay psychiatric ward and in the early months following their move to the community. During the period before the move, staff and residents colluded to prevent residents' active involvement in the preparations. Immediately following the move, staff seemed unsure of their roles and value, and residents expressed continued uncertainty about their identity as psychiatric patients versus members of the community. The results were interpreted in terms of the threat that transition from hospital to community represents to the defensive social structures developed by each group to cope with the anxieties inherent in their position. For the residents, this threat is to their already ambiguous identities as neither "mad" nor "normal". For the staff, it means greater intimacy with the residents and raises expectations of improvement that may not be fulfilled. PMID- 14529078 TI - A comparison of the quality of life of severely mentally ill people in UK & German samples. AB - The improvement of the quality of life of people with a severe mental illness is a key policy objective and an important outcome for clinical services. Drawing on cases assessed using the Lancashire Quality of Life Profile and its German translation (The Berliner Lebensqualitatprofil), this paper explores the relationship between personal characteristics, objective well being, subjective well being and overall well being. These variables are compared in two large data sets of people with severe mental illness, one from the UK (n = 1279) and the other from Germany (n = 386). The comparison shows that UK cases have significantly lower subjective well-being in almost all life domains (except safety, living situation and employment). UK cases reported slightly but not significantly higher levels of satisfaction with employment but German cases are more often employed than their UK counterparts. The German samples reported substantially better subjective well-being ratings for health, finances, family, leisure and social life. Exploration of the predictors of overall well-being shows that in both countries depression has the effect of reducing subjective well-being scores, except in relation to work (both samples), religion (UK), finance and safety (Germany). Regression analysis confirms that age, depression and objective circumstances make a small contribution to overall well-being but that subjective ratings in individual life domains make the major contribution. The most important individual predictors of overall well-being for the two samples combined include being a victim of crime, depression and satisfaction with leisure, work, health and mental health, family, living situation, finance and social contacts. Factor analysis indicates that the variance in global well being explained in both samples combined is 36% (31% in the German samples and 38% in the UK sample). PMID- 14529079 TI - Housing needs of consumers of mental health services in rural New South Wales, Australia. AB - A survey of the housing needs of 101 people with mentally illness treated by the Central Western Area Mental Health Service were compared with data from the 1994 Australian Housing Study (Australian Bureau of Statistics). The results indicate that people with mental illness want housing similar to Australians in general. However, unemployment or very low incomes may affect their ability to realise their housing choices. Despite very low incomes most in the survey felt their rent was affordable. Preferences for housing types and factors relating to housing choice are described. The results are discussed in relation to the importance of housing in maintaining mental health. PMID- 14529080 TI - Stress in farmers. AB - This study aims to document the nature and extent of occupational stress in farming. A postal survey of farmers in the South West of England, was undertaken and results indicate high levels of occupational stress in farming families. 35% of respondents scored positively on the General Health Questionnaire (GHQ) with female respondents showing significantly higher scores than males. A significant proportion of respondents also showed elevated levels of anxiety and depression as measured by the Hospital Anxiety and Depression Scale (HAD). Indices of psychological distress were correlated with ill health and family problems though the most common sources of perceived stress were coping with new legislation, the amount of paper work and media criticism. People who reported a greater number of stressors scored higher on all 3 measure of psychological distress. PMID- 14529081 TI - Evaluation of postpartum depression using the Edinburgh postnatal depression scale in evaluation of postpartum depression in a rural community in India. PMID- 14529082 TI - Mosquito burden and impact on the poor: measures and costs for personal protection in some communities in Thailand. AB - To gauge the extent of mosquito problems and their impact on local people in Thailand, a simple questionnaire was designed consisting of 6 questions with multiple choices to be answered in 4 different communities in Thailand in 1998 and 1999. Mosquito biting activity was noted often by respondents. They reported that mosquitoes bit both night and day, and that the insects were abundant both in the dry and the rainy seasons. In all 4 communities, a large proportion of the residents used bed nets, mosquito coils, and aerosol sprays for personal protection; vaporizing mats and repellents were used sparingly. The cost of such measures amounted to dollars 4 to dollars 25 per year per household. For most of the residents, this represented a substantial proportion of their income, and was proportionally greater than the average cost of organized mosquito control in developed countries. This suggests that instituting organized local vector control programs would be cheaper and more effective than the individual use of personal protectants that do not reduce mosquito numbers. An assessment of the available products stocked in neighborhood stores and supermarkets for personal protection was made. A variety of insecticidal aerosols, mosquito coils, liquid sprays, vaporizing mats, and vaporizing liquids was stocked. This ample supply of household insecticides lends support to the preferred methods of protection reported by the respondents. The active ingredients in most of the formulations were synthetic pyrethroids, although a few contained dichlorvos, propoxur, and a few other compounds. Mosquito coils, the most preferred products used by the poor, were evaluated for efficacy, and were found to provide a reduction of 72 96% in landing-biting rates in controlled experiments. PMID- 14529083 TI - False detection of Plasmodium falciparum sporozoites in Anopheles marshallii group mosquitoes. AB - A study was conducted to determine the role of members of the Anopheles funestus group in malaria transmission in the Mpumalanga Province, in the northeastern region of South Africa. Female anopheline mosquitoes were collected between January 1996 and November 1997 by means of human landing catches and tested for salivary gland Plasmodium falciparum infections by means of the enzyme-linked immunosorbent assay (ELISA) method with PF2A10 antibodies. Infection rates from April and May 1997 collections were 3.73% and 19.4%, respectively. None of the nonimmune collectors became infected with malaria. The ELISA-positive mosquitoes were tested with a polymerase chain reaction (PCR) malaria detection assay based on sequence variation present in the small subunit ribosomal RNA gene. Only 1.09% of ELISA-positive mosquitoes were PCR-positive for malaria. Initially, all mosquitoes were assumed to belong to the An. funestus group but subsequent molecular taxonomy showed this assumption to be false. The use of a single-strand conformation polymorphism (SSCP) assay revealed only 1 member of the An. funestus group, An. rivulorum. All other specimens produced banding patterns not seen before. Those samples were identified morphologically as An. demeilloni and An. marshallii s.l. These 2 species are not recognized malaria vectors and thus it is possible that the ELISA results are misleading. PMID- 14529084 TI - Mosquito vector control and biology in Latin America--an eleventh symposium. AB - The 11th annual Latin American symposium presented by the American Mosquito Control Association (AMCA) was held as part of the 67th Annual Meeting in Dallas, TX, in February 2001. The principal objective, as for the previous 10 symposia, was to promote participation in the AMCA by vector control specialists, public health workers, and academicians from Latin America. This publication includes summaries of 45 presentations that were given orally in Spanish or presented as posters by participants from 8 countries in Latin America. Topics addressed in the symposium included results from chemical and biological control programs and studies; studies of insecticide resistance; and population genetics, molecular, ecological, and behavioral studies of vectors of dengue (Aedes aegypti), malaria (Anopheles albimanus and Anopheles aquasalis), leishmaniasis (Lutzomyia), murine typhus, and Chagas' disease (Triatoma). Related topics included biology and control of Rhodnius, scorpions, Loxosceles spp., Chironomus plumosus, and Musca domestica. PMID- 14529085 TI - Ecological distribution of mosquito larvae of the Anopheles punctulatus group on Niolam (Lihir) Island, Papua New Guinea. AB - We surveyed the larval habitats of members of the Anopheles punctulatus group of mosquitoes on Niolam (Lihir) Island, Papua New Guinea. Identification of this group was undertaken by polymerase chain reaction-restriction fragment length polymorphism analysis of the amplified internal transcribed spacer unit 2 of rDNA, because morphologic separation of member species is unreliable. The most widespread malaria vector species and their most common larval habitats were identified to aid source-reduction programs for malaria control. The most ubiquitous species was An. punctulatus, followed by An. farauti no. 2. then An. farauti s.s. Anopheles punctulatus has increased relative to An. farauti s.l. since the start of development projects on Lihir Island. The most common larval habitats were shallow temporary pools with clay substrate and with plants or floatage. These habitats, mostly encountered alongside poorly drained roads, may be increased by development projects. PMID- 14529086 TI - Characterization of male and female wingbeat frequencies in the Anopheles quadrimaculatus complex in Mississippi. AB - Classifying individual mosquitoes of the Anopheles quadrimaculatus species complex to species has always been time consuming and complex, involving genetic analysis and profiling. We characterized the wingbeat frequencies of the 3 species of the Anopheles quadrimaculatus complex (An. quadrimaculatus, An. smaragdinus, and An. maverlius) that occur in Mississippi to determine if this character could successfully distinguish between individuals of the 3 species. Wingbeats of females varied from 320 to 480 beats/sec, but no significant differences in wingbeat frequencies were found among females from 3 populations of the 3 species. Wingbeats of males were higher than those of females (500-770 beats/sec), and the mean wingbeat frequency of An. maverlius was significantly different from the means of An. quadrimaculatus and An. smaragdinus, although overlap occurred at the individual level. Such overlap precludes use of wingbeat frequencies as an identification mechanism, and indicates that, at least for the An. quadrimaculatus complex, wingbeat frequencies are not involved in mate recognition. PMID- 14529087 TI - Laboratory and field evaluations of the insect repellent 3535 (ethyl butylacetylaminopropionate) and deet against mosquito vectors in Thailand. AB - The insect repellents 3535 (ethyl butylacetylaminopropionate or IR3535) and deet (N,N-diethyl-3-methylbenzamide) were prepared as 20% solutions in absolute ethanol and evaluated for repellency against many mosquito species in Thailand under laboratory and field conditions using human subjects. In the laboratory, 0.1 ml was applied per 30-cm2 of exposed area on a volunteer's forearm (0.66-0.67 mg active ingredient [AI]/ cm2), whereas in the field, volunteers' legs (from knee to ankle, with a surface area of about 712-782 cm2) were treated with 3 ml per exposed area (0.76-0.84 mg AI/cm2). In the laboratory, both IR3535 and deet showed equal repellency (P > 0.05) for 9.8 and 9.7 h against Aedes aegypti, for 13.7 and 12.7 h against Culex quinquefasciatus, and for 14.8 and 14.5 h against Cx. tritaeniorhynchus, respectively. Anopheles dirus was significantly less sensitive to IR3535 than to deet (P < 0.05), with a mean protection time of 3.8 and 5.8 h, respectively. Under field conditions, both IR3535 and deet provided a high degree of protection against various mosquito vectors ranging from 94 to 100% during the test periods. Both repellents provided a high level of protection for at least 8 h against Ae. albopictus and for at least 5 h against Cx. gelidus, Cx. tritaeniorhynchus, Cx. quinquefasciatus, Mansonia dives, Ma. uniformis, Ma. annulata, Ma. annulifera, Anopheles minimus, and An. maculatus. This study clearly documents the potential of IR3535 for use as a topical treatment against a wide range of mosquito species belonging to several genera. PMID- 14529088 TI - Light, carbon dioxide, and octenol-baited mosquito trap and host-seeking activity evaluations for mosquitoes in a malarious area of the Republic of Korea. AB - Two field trials for commercially available and experimental mosquito traps variously baited with light, carbon dioxide, octenol, or combinations of these were evaluated in a malarious area at Paekyeon-Ri near Tongil-Chon (village) and Camp Greaves, Paju County, Kyonggi Province, Republic of Korea. The host-seeking activity for common mosquito species was determined using hourly aspirator collections from a human- and propane lantern-baited Shannon trap. The total number of mosquitoes and number of each species captured during the test were compared using 8 x 8 and 5 x 5 Latin square designs based on trap location. Significant differences were observed for the total number of mosquitoes collected in the 8 x 8 test, such that counterflow geometry (CFG) with CO2 > or = CFG with CO2 and octenol > or = Shannon trap > or = Mosquito Magnet with octenol > American Biophysics Corporation (ABC) light trap with light, CO2 (500 ml/min), and octenol > or = ABC light trap with light and dry ice > or = ABC light trap with light and CO2 > ABC light trap with light only. A concurrent 5 x 5 test found significant differences in trap catch, where Mosquito Magnet with octenol > New Jersey light trap > or = EPAR Mosquito Killer with CO2 > or = ABC light trap with light and dry ice > Centers for Disease Control (CDC) light trap (manufactured by John W. Hock) with light and octenol. Significant differences in trap catch were noted for several species including: Aedes vexans, Anopheles sinensis, An. yatsushiroensis, An. lesteri, Culex pipiens, and Cx. orientalis. Traps baited with octenol captured significantly fewer Cx. pipiens than those not baited with octenol. Likewise, no Cx. orientalis were captured in octenol-baited traps. Host-seeking activity showed a similar bimodal pattern for all species captured. Results from these field trap evaluations can significantly enhance surveillance efforts. Significantly greater numbers of mosquitoes were captured with mosquito traps using counterflow technology (e.g., Mosquito Magnet and CFG traps) when compared to standard light and carbon dioxide-baited traps. Additionally, field evaluations demonstrate that various traps can be utilized for isolation and detection of arboviruses and other pathogens. PMID- 14529089 TI - Effect of some animal feeds and oviposition substrates on Aedes oviposition in ovitraps in Cairns, Australia. AB - Animal feed pellets containing lupin seed or alfalfa were added to ovitraps set in Cairns, Australia. Although they collected fewer Aedes eggs than Centers for Disease Control enhanced ovitraps, they did outperform tap water alone. A wooden tongue depressor collected comparable number of Aedes eggs as a Masonite board and seed germination paper. PMID- 14529090 TI - Update on the biology of Triatoma dimidiata Latreille (Hemiptera: Reduviidae) under laboratory conditions. AB - Feeding time, postfeeding defecation delay, and life cycle for each stage of a cohort of recently colonized Mexican Triatoma dimidiata were evaluated, and results were compared to existing published information on this species. Seventy five nymphs (41.7%) completed a cycle with an average time from N-I to adult of 142 +/- 64 days. The average span in days for each stage was 20.2 for N-I, 17.9 for N-II, 10.1 for N-III, 43.6 for N-IV, and 55.1 for N-V. First-stage nymphs had the highest mean feeding time (25 min) and the longest postfeeding defecation delay (45 min). Differences among biological data from previous studies and the present study confirm the importance of conducting research on the behavior of the indigenous triatomine species from various countries. PMID- 14529091 TI - Mechanisms of metastasis as related to receptor tyrosine kinases in small-cell lung cancer. AB - Small-cell lung cancer (SCLC) is an aggressive, malignant neoplasm with a 5-year survival of less than 10%. This poor survival rate is related to a high propensity for recurrence and a high rate of metastases. Metastases initially occur in the lymph nodes and thereafter in other organs such as the lung itself, liver, adrenal glands, brain, bone, and bone marrow. The mechanisms of metastases have been better understood recently and are described in this review. Receptor tyrosine kinases (RTKs) have been identified as important therapeutic targets in non-small-cell lung cancer (such as the EGF-receptor). We have begun to identify RTKs in SCLC and have shown that c-Kit and c-Met are expressed and functional in SCLC. RTKs have also been shown to be important in the metastasis of cancer cells. The roles of RTKs in the mechanism of metastasis are detailed in this review, with special emphasis on downstream signal transduction from RTK signaling. PMID- 14529093 TI - c-Myc-induced genomic instability. AB - c-myc is one of a small family of proto-oncogenes that do not require mutation to contribute to neoplastic transformation. Instead, the deregulated expression of the oncoprotein at even modest levels is sufficient to initiate this process. The transforming activity of c-Myc is generally thought to lie in its ability to modulate the expression of a series of genes, among them certain proliferation promoting genes. In reality, c-Myc is a multifunctional protein that also affects the stability of the genome. In this review, we summarize the growing evidence that deregulated c-myc expression generates genomic instability by initiating gene amplification (both intra- and extra-chromosomally), gene rearrangements, and karyotypic instability. Cancer is a disease of impaired genomic stability, to which c-Myc contributes during its initiation and progression through the induction of genomic instability in critical genes. Myc thus acts as a structural modifier of the genome and as a promoter of neoplastic transformation. PMID- 14529092 TI - Binding of alpha5 monoclonal antibody to cell surface alpha5beta1 integrin modulates MMP-2 and MMP-7 activity in B16F10 melanoma cells. AB - The integrin multigene family, comprising at least 21 distinct heterodimeric complexes, includes receptors for all major extracellular proteins such as fibronectin, laminin, vitronectin, and collagens. These receptors play important roles in various physiological processes. Matrixmetallo-proteinases (MMPs) are a family of zinc-dependent endopeptidases that degrade the extracellular matrix (ECM) components in physiological conditions. MMPs can degrade all structural components of the ECM, which is one of the critical aspects of tumor cell invasion. The role of matrilysin (MMP-7) in the progression of various carcinomas was recently addressed. Evidence of interplay between integrin function and ECM matrix integrity has come from experiments showing that integrin-ECM contacts can regulate expression and function of MMPs. In this communication, we report on an interesting interrelationship between cell surface alpha5beta1 integrin (fibronectin receptor) and MMPs (MMP-2 and MMP-7). RESULTS: The zymographic analysis showed that ligation of cell surface alpha5beta1 integrin by alpha5 monoclonal antibody leads to the expression and activation of MMP-2 and MMP-7 in B16F10 melanoma cells. The immunoblot confirmed the tyrosine phosphorylation of FAK in B16F10 cells grown in presence of alpha5 monoclonal antibody, indicating the transduction of signal via FAK. The immunocytochemical study demonstrated that alpha5beta1 integrin stimulation causes rearrangement of actin fibers and its accumulation in focal adhesion sites. CONCLUSION: alpha5beta1 integrin induced expression and activation of MMP-2 and -7 indicates the role of tumor cell surface integrin receptor in the modulation of MMPs and, thereby, the invasive property of tumor cells. PMID- 14529094 TI - Human immunodeficiency virus type 1 pharmacogenomics in clinical practice: relevance of HIV-1 drug resistance testing (Part 1). AB - Throughout most of the past century, physicians could offer patients no treatments for infections caused by viruses. The experience with treatment of infection by human immunodeficiency virus (HIV) has changed the way healthcare workers deal with viral infections and has triggered a growing rate of discovery and use of antiviral agents, the first fruits of the expanding genomics revolution. HIV treatment also provides an informative paradigm for pharmacogenomics because control of infection and its consequences is limited by the development of viral drug resistance and by host factors. This report summarizes studies published to date on the significance of testing of HIV-1 resistance to antiretroviral drugs. The only Food and Drug Administration approved kit for HIV drug resistance testing by genotypic sequencing is commercially available through Visible Genetics, Inc. Genotyping sequencing alone is most likely an adequate test to assist in the therapeutic decision-making process for previous regimen failure, for treatment-naive patients in areas of high prevalence of transmitted resistant virus, and for pregnant women. However, in exceptional cases of highly complex mutation patterns and extensive cross resistance, it may be useful to obtain a phenotype test, because that result may more easily identify drugs to which virus is least resistant. There are no published clinical trials results on the usefulness of the so-called virtual phenotype over genotypic sequencing alone. Not only has the paradigm of viral pharmacogenomics in the form of HIV genotypic sequencing been useful in treating other viral diseases, but it is also important to the real-life implementation of the growing discipline ofgenomics or molecular medicine. The application of this paradigm to the thousands of potential therapeutic targets that have become available through the various human genome projects will certainly gradually change the landscape of diagnosis and management of many diseases, including cancer. PMID- 14529095 TI - Benefits of swimming in asthma: effect of a session of swimming lessons on symptoms and PFTs with review of the literature. AB - A study involving eight children with moderate persistent asthma was undertaken to determine whether standard swimming lessons improved symptoms and pulmonary function tests (PFTs) in asthmatic children. Five children ages 7-12 years old with moderate persistent asthma were randomized to a swimming lesson group (5- to 6-week session) and three to a control group. Both groups completed pre- and poststudy period PFTs and symptom questionnaires. Swimming lessons did not produce a significant change in asthma symptoms or PFTs. Review of previous literature found that swimming has been shown to have definite benefits in improving cardiorespiratory fitness in asthmatic children. Swimming has been shown to be less asthmogenic than other forms of exercise. Some studies have also shown improvement in asthma symptoms in children participating in exercise programs. PMID- 14529096 TI - Prevalence and correlates of asthma in the Puerto Rican population: Behavioral Risk Factor Surveillance System, 2000. AB - The 2000 Behavioral Risk Factor Surveillance System (BRFSS) showed that Puerto Rico had the highest self-reported prevalence of asthma. Our objective was to estimate the self-reported prevalence of asthma among different population subgroups and determine its correlates in Puerto Rican adults as reported by the BRFSS. The BRFSS data gathered during 2000 were analyzed. To determine factors associated with self-reported prevalence of asthma, a simple unconditional logistic regression model was employed; then, to estimate adjusted weighted prevalence odds ratios, a multiple unconditional logistic regression model was used. The self-reported weighted prevalence of ever having asthma among Puerto Rican adults was 15.9% (14.8%-16.9%). Asthma prevalence was significantly higher in the following population subgroups: females (18.8%), educational attainment > 12 years (18.4%), having health coverage (16.3%), and obesity (21.0%). Asthma prevalence did not differ among age groups, region of residence, annual income, smoking at least 100 cigarettes in entire life, and physical activity. Almost half (45.6%) of asthmatics reported having children affected with the condition. The prevalence of asthma in any children of the interviewed was 33.2%, 51.3% were receiving treatment, and 30.6% and 24.3% reported having one to three visits to emergency departments and hospital admissions, respectively, resulting from asthma last year. Based on the logistic regression model, the following factors were significantly associated with asthma: sex, high educational attainment, health coverage and obesity. Consistent with previous studies in Puerto Ricans living in the mainland, a higher than expected prevalence of asthma was observed. The possibility of a genetic-environment interaction deserves further investigation. PMID- 14529097 TI - Evaluation of parental preference for the treatment of asthmatic children aged 6 to 11 years with oral montelukast or inhaled cromolyn: a randomized, open-label, crossover study. AB - The objective of this study was to evaluate parental preference for the treatment of asthmatic children with oral montelukast sodium or inhaled cromolyn sodium. Additionally, we wanted to compare the two drugs in terms of patient preference for treatment, patient and parent satisfaction with treatment, frequency of inhaled albuterol use, adherence to treatment, and safety. This was a 12-week randomized, open-label, crossover study conducted in 42 primary care and asthma/allergy specialty centers in the United States. Three hundred thirty-three asthmatic patients, ages 6 to 11 years, who had a forced expiratory volume in 1 second (FEV1) of 60%-85% (inclusive) of predicted value with > or = 12% reversibility after administration of an inhaled beta-agonist and who used albuterol on at least 7 of the last 14 days of the run-in period. After a 2- to 3 week run-in period, patients were randomized either to 4 weeks of montelukast (5 mg chewable tablet once daily) followed by a 2-week washout period, then 4 weeks of cromolyn (two puffs 4 times daily from a metered-dose inhaler) or to the reverse sequence. More parents preferred montelukast (87%) than cromolyn (12%; p < 0.001). More patients preferred montelukast (82%) than cromolyn (17%; p < 0.001). Daily albuterol use (puffs/day) was reduced by 38% during montelukast therapy vs. 23% during cromolyn therapy. Seventy-eight percent of patients reported being highly adherent to montelukast therapy compared with 42% to cromolyn therapy (p < 0.001). Fewer patients receiving montelukast discontinued because of asthma exacerbation (1.0% vs. 5.0%, respectively), and fewer patients reported worsening asthma while receiving montelukast (3.5% vs. 7.5%, p = 0.036). Parents' and patients' preference, parents' and patients' satisfaction, and patients' adherence to therapy were all significantly better with oral montelukast compared with inhaled cromolyn. Beta-agonist use was decreased when taking montelukast, which was safe and well-tolerated. PMID- 14529098 TI - Efficacy response of inhaled HFA-albuterol delivered via the breath-actuated Autohaler inhalation device is comparable to dose in patients with asthma. AB - Handling difficulties, such as poor coordination of actuation and inhalation, are common in patients using press and breathe (P&Bs) metered-dose inhalers to administer asthma medication. Although spacers can help overcome some difficulties, the cumbersome nature of these devices often detracts from their use for the administration of rescue medications, where portability is important. This randomized, placebo-controlled, multicenter, crossover study investigated the efficacy, dose-response and safety of HFA-albuterol delivered via a breath actuated Autohaler inhalation device in comparison with the same medication delivered using a conventional P&B device. In total, 39 patients received six study treatments in a random sequence at clinic visits separated by 2-7 days: 2 puffs from a HFA-placebo Autohaler; 1, 2, or 4 puffs from a HFA-albuterol Autohaler; I or 2 puffs from a HFA-albuterol P&B. Both active inhalers delivered 90 microg albuterol base equivalent/actuation from the actuator. The change from baseline in forced expiratory volume in 1 s (FEV1) and the area under the FEV1 curve (FEV1 AUC) were significantly greater than placebo for all active treatment groups (p < or = 0.01) and were suggestive of a dose response for each inhaler. Examination of the pooled slope of the dose responses for the Autohaler and P&B using Finney's Parallel Line Bioassay Methodology found a highly statistically significant relationship indicating the equivalence of the two inhalers on both parameters (p < or = 0.002). The relative potency of the two inhalers was 0.8 (95% CI: 0.47, 1.46) for the mean change from baseline in FEV1 and 0.9 (95% CI, 0.56, 1.48) for the change from baseline in FEV1 AUC. There was also a trend toward an increase in the mean percentage change from baseline in FEV1 as the number of puffs increased for both inhalers. Furthermore, there were no significant differences between the treatment groups with regard to time to onset of bronchodilator effect and the duration of effect was significantly greater than placebo (p < or = 0.01) in each of the active groups. Adverse events were generally mild to moderate in nature and were of similar incidence (< or = 18% of patients) in each group. This study demonstrates a dose-response for HFA albuterol on bronchodilation using both the Autohaler and P&B devices and illustrates that, in patients with good coordination of inhalation with actuation, the efficacy and safety of the two inhalers is similar at equivalent doses. PMID- 14529099 TI - Traditional oriental herbal medicine, Bakumondo-to, suppresses vagal neuro effector transmission in guinea pig trachea. AB - OBJECTIVE: Bakumondo-to (Maimendong tang) is a traditional oriental herbal medicine that has been used as an antitussive agent. We previously demonstrated that Bakumondo-to attenuates airway hyperresponsiveness induced by ozone. However, the mechanism(s) responsible for this effect remains unclear. In the present study, we examined the mechanism whereby Bakumondo-to inhibits ozone induced airway hyperresponsiveness. First, we examined the effect of Bakumondo-to on prostanoids production, which are key mediators to airway hyperresponsiveness after ozone exposure. Second, we studied its effects on the vagal neuroeffector transmission, because vagal nerve is likely to play an important role in airway hyperresponsiveness after ozone. METHODS: We measured the effects of Bakumondo-to on the concentrations of prostanoids in bronchoalveolar lavage fluid before and after ozone. We evaluated the effects of Bakumondo-to on the contraction of guinea pig tracheal smooth muscle evoked by electrical field stimulation (EFS) or the exogenous application of acetylcholine (ACh). Isometric tension of tracheal strips was measured in the presence of indomethacin (10(-6) M) and of guanethidine (10(-6) M). RESULTS: Ozone caused significant increase in prostaglandin E2 (PGE2) and thromboxane B2 (TXB2); however, Bakumondo-to did not affect the increase in these prostanoids. Bakumondo-to (0.01 mg/mL-1 mg/mL) significantly suppressed the contraction evoked by EFS, but did not affect the ACh-evoked contraction, indicating that Bakumondo-to suppressed tracheal smooth muscle contraction pre-junctionally. CONCLUSION: These results suggest that the mechanism by which Bakumondo-to inhibits airway hyperresponsiveness depends on inhibiting the release of acetylcholine from vagal nerve terminals. PMID- 14529100 TI - Formoterol delivered via the dry powder Aerolizer inhaler versus albuterol MDI and placebo in mild-to-moderate asthma: a randomized, double-blind, double-dummy trial. AB - The objectives of this study were to compare the efficacy and tolerability of twice-daily formoterol dry powder 12 microg and 24 microg (Foradil) delivered via Aerolizer inhaler with four times daily albuterol (salbutamol) 180 microg delivered via metered dose inhaler (MDI) and placebo. A total of 554 adolescents and adults (ages 12-75 years) with mild-to-moderate asthma were randomized to this 12-week, multicenter, double-blind, double-dummy, placebo-controlled, parallel-group study. Twelve-hour spirometry measurements were taken at weeks 0, 4, 8, and 12. A total of 484 patients completed the study (122, 116, 127, and 119 given formoterol 12 microg, formoterol 24 microg, albuterol, and placebo, respectively). For the primary efficacy variable, the forced expiratory volume in 1 second (FEV1), both formoterol 12 microg and 24 microg were statistically superior to placebo at all time points on all test days (p < or = 0.017) and to albuterol at most time points on all test days (p < or = 0.001). The onset of improvement in FEV1 was rapid, with 15% increase within 5 min in 57%, 71%, and 65% of formoterol 12 microg, formoterol 24 microg, and albuterol patients, respectively. Formoterol was also superior to placebo and albuterol in terms of secondary efficacy variables: FEV1 area under the curve, percentage of predicted FEV1, forced vital capacity and forced expiratory flow, asthma symptom scores, and peak expiratory flows. In conclusion, both formoterol doses were superior to placebo in all lung function measurements. Overall, compared with albuterol, both formoterol doses produced superior bronchodilation. Formoterol and albuterol were safe and well-tolerated. PMID- 14529101 TI - Adrenal function as assessed by low-dose adrenocorticotropin hormone test before and after switching from inhaled beclomethasone dipropionate to inhaled fluticasone propionate. AB - Low-dose adrenocorticotropin hormone (ACTH) tests (0.5 microg/L 73 m2) were done before and after switching from inhaled beclomethasone dipropionate to inhaled fluticasone propionate in 12 patients 33-77 years old who had mild-to-severe asthma to compare the effects of these drugs on adrenal function. Low-dose ACTH tests were performed after the subjects had received inhaled beclomethasone dipropionate (200-900 microg/day) for at least 12 wk. Treatment was then switched to inhaled fluticasone propionate (200-600 microg/day) for at least 12 wk, and a second low-dose ACTH test was done. Pulmonary function was assessed on the basis of peak expiratory flow rate (PEFR, % of predicted value). After switching treatment, the daily dose of inhaled corticosteroid decreased by about 40%. Basal serum cortisol and ACTH levels were similar with both treatments. The adrenal response, as assessed by incremental rise in the serum cortisol level (peak minus basal) after ACTH challenge, improved significantly (5.6-7.9 microg/dL, p < 0.01) after switching to fluticasone. All three patients who had lower serum cortisol levels during beclomethasone treatment than during fluticasone treatment showed improvement in both the peak cortisol level and the incremental rise in cortisol. Mean morning and evening PEFRs significantly increased after switching from beclomethasone to fluticasone (morning: 71.2 to 76.0%, p < 0.01; evening: 67.3 to 72.1%, both p < 0.05). The diurnal variation of PEFR significantly decreased from 10.9% to 8.3% after switching treatment (p < 0.01). We conclude that switching from beclomethasone to fluticasone reduces the risk of adrenal dysfunction associated with inhaled steroids and improves pulmonary function. PMID- 14529102 TI - Evaluation of an education program for elementary school children with asthma. AB - To evaluate the effectiveness of a comprehensive asthma management education program for 7- to 12-year-old children with asthma, entitled Roaring Adventures of Puff (RAP), 18 elementary schools in Edmonton were randomized to intervention and control groups. Participating in the program were 76 students with asthma in the intervention schools and 86 in the control schools. Children in the intervention schools had statistically significant improvements in unscheduled doctor visits, missed school days, moderate-to-severe parent rating of severity, severity of shortness of breath, limitations in the kind of play, and correct use of medications. Unscheduled doctor visits and missed school days were the only significant improvements in the control group; however, improvements were about half that of the intervention group. The results showed that a comprehensive, school-based asthma education program is feasible and improves outcomes. PMID- 14529103 TI - Prevalence of childhood asthma and associated morbidity in Los Angeles County: impacts of race/ethnicity and income. AB - Racial/ethnic and socioeconomic disparities in the prevalence of childhood asthma have been reported nationally but few population-based studies in local and regional settings have been reported. To assess variation in the prevalence of childhood asthma and associated morbidity across race/ethnic and income groups in the Los Angeles County population, we analyzed data on a random sample of 6004 children (< or = 17 years old) enrolled in a countywide health survey from September 1999 through April 2000. The prevalence of childhood asthma was highest in blacks (15.8%), intermediate in whites (7.3%) and Asians (6.0%), and lowest in Latinos (3.9%; p < 0.001). These differences persisted after controlling for income, measures of health care access, and other covariates. Asthma prevalence was inversely related to income in all racial/ethnic groups except Latinos from Spanish-speaking households. Among children with asthma, blacks and Latinos were more likely than whites to report asthma-related limitations in physical activity and need for urgent medical services. These findings indicate marked disparities in asthma prevalence and related morbidity in this large urban child population and highlight the importance of efforts to identify high-risk subpopulations for focused prevention and treatment interventions. PMID- 14529104 TI - Risk for asthma in 1-year-old infants residing in New York City high-risk neighborhoods. PMID- 14529105 TI - Prevalence of asthma and related symptoms in Sivas, Central Anatolia. AB - BACKGROUND: In our clinical experience, asthma is an important health problem in our region, but we did not know its prevalence. OBJECTIVE: The main objective of our study was to assess the prevalence of asthma in Sivas, a city in Central Anatolia. METHODS: A cross-sectional study was performed using a screening questionnaire adopted from European Community Respiratory Health Survey (ECRHS). A total of 5448 adults of both genders between 20 and 107 yr of age living in Sivas, a city in the central region of Anatolia in Turkey, participated in the survey. Of the cohort, 2691 were men (49.4%) and 2757 (50.6%) were women. RESULTS: The mean age was 38.2 yr (SD = 12.7 yr), almost half of the study population was at or younger than 40 yr of age. The prevalence of wheezing in the last 12 months, diagnosis of asthma, asthma attack in the last 12 months, and use of asthma medicine were 20.9%, 4.5%, 4.9%, and 3.4%, respectively. Awakening with chest tightness, with shortness of breath, or with cough were reported as 14.2%, 14.8%, and 22.7%, respectively. The prevalence of respiratory symptoms related to asthma was statistically higher in women than that of men (p = 0.000, OR 1.346, 95% CI: 1.228-1.475). Those who had asthma/allergic symptoms in their family members were significantly younger than others (p: 0.001). CONCLUSIONS: The results of this study demonstrate that symptoms suggestive of asthma are quite common and constitute a major health problem in Sivas, Turkey. This study also showed that, despite a high rate of reported symptoms exists, the rate of diagnosis and treatment of asthma is low among the adult population in Sivas. PMID- 14529106 TI - Elevated asthma and indoor environmental exposures among Puerto Rican children of East Harlem. AB - OBJECTIVE: East Harlem in New York City, a community with a large Puerto Rican population, has among the highest rates of asthma hospitalizations and mortality in the United States, but it is not known if the high rates are related to the ethnic composition, environmental or community factors, or if the higher rates reflect differentials in access to appropriate asthma care. A survey was conducted to: (a) estimate the prevalence of current asthma by ethnicity among school-age children, (b) assess indoor environmental risk factors for childhood asthma, and (c) assess health care utilization and school absences associated with childhood asthma. DESIGN: A cross-sectional survey of parents of elementary school children, using a self-administered questionnaire with a 12-month recall on asthma symptoms based on the International Study of Asthma and Allergies in Childhood. SETTING: Two public elementary schools in East Harlem (n = 1615 students 5-12 years of age). RESULTS: Among the 1319 respondents (response rate 82%), the prevalence for current asthma (doctor or nurse diagnosis at any time plus wheezing in the past 12 months) was 23%. Puerto Rican children had a prevalence of 35%. Puerto Rican children reported both higher symptomatic frequencies and higher rates of physician diagnosis. Living in a home where cockroaches, rats, or mice had been seen in the past month and with a dust enhancing heating system also was associated with having asthma, regardless of ethnicity. Compared with other children with asthma, Puerto Rican children with asthma were more likely to live in homes where rats or mice had been seen in the past month. Regardless of ethnicity, children with more frequent, more severe asthma symptoms and incomplete asthma action plans were more likely to have visited the emergency department in the past year. Puerto Rican children were more likely to have missed school because of their asthma in the past year. CONCLUSION: The prevalence of current asthma was significantly higher among Puerto Ricans, who had higher symptomatic frequency and greater diagnosis rates. Although all children with asthma in the East Harlem study appear to be sensitive to selected indoor environmental risk factors, only Puerto Rican children with asthma appear to be sensitive to the presence of rodents in their buildings. However, their higher school absence rate suggests problems with routine asthma management that could be addressed by improved medical management, programs to help parents manage their children's asthma, or school staff assistance with medications. PMID- 14529107 TI - Exposure of children to environmental tobacco smoke (ETS) and its association with respiratory ailments. AB - OBJECTIVES: To assess the impact of domestic passive smoking (cigarettes or narghile) on the development of respiratory ailments among children ages 10-15 years. METHODS: Students were recruited from five private schools in Beirut, and information on demographic, in-home smoking, and students' respiratory tract illnesses (cough, wheezing, runny nose, or nasal congestion) were collected from each participant. RESULTS: Of 625 students surveyed, 438 (70.1%) had at least one individual smoking at home. Compared with the nonexposed group, the odds ratio of having respiratory illness for children exposed to narghile or cigarette smoke were 2.3 (95% Confidence Interval [CI] 1.1-5.1) and 3.2 (95% CI 1.9-5.4), respectively. CONCLUSIONS: In addition to the well-established effect of smoking, domestic passive smoking of the misconceived "innocuous" habitual smoking device, narghile, is associated with significant respiratory health ailments. PMID- 14529108 TI - Factors influencing parent reports on quality of life for children with asthma. AB - Children and parents often differ in their perceptions of a complex disease such as asthma. This ancillary study of children with mild to moderate asthma that was conducted at four of the eight clinics in the Childhood Asthma Management Program had two aims: (1) to relate quality of life to asthma symptoms, sociodemographic characteristics, child psychosocial adjustment and family social support and (2) to relate agreement between child- and parent-reported quality of life to child age. For this study participants completed the Caregiver's Pediatric Asthma Quality of Life Questionnaire (C-PAQLQ), Pediatric Asthma Quality of Life Questionnaire, and a battery of psychosocial health outcomes questionnaires at the 12-month follow-up. The average asthma symptom score from diary cards for the 2 weeks before the visit was also collected. Parent-reported family burden was the strongest correlate of C-PAQLQ scores. Although the average asthma symptom score was not associated with parent-reported quality of life, the study had low power to detect a relationship because of the few episodes of asthma reported during the assessment period. The correlation between child and parent reported quality of life improved with increasing age of the child, suggesting increased child-parent agreement about quality of life outcomes was associated with increasing age of the child. Family characteristics, such as degree of cohesion among family members, are not associated with quality of life ratings. PMID- 14529109 TI - Perceiving prosody from the face and voice: distinguishing statements from echoic questions in English. AB - We examined the processing of potential auditory and visual cues that differentiate statements from echoic questions. In Experiment 1, four natural speech statement-question pairs were identified by participants, and then analyzed to determine which characteristics were ecologically valid. These characteristics were tested in subsequent experiments to determine if they were also functionally valid. In Experiment 2, the characteristics of the most discriminable utterance pair were successfully extended to the other utterance pairs. For Experiment 3, an auditory continuum (varying in F0, amplitude, duration) was crossed with a visual continuum (varying in eyebrow raise, head tilt), using synthetic speech and a computer-animated head. Participants judged five levels along each of these two speech continua between a prototypical statement and prototypical question, in an expanded factorial design. Experiments 4 and 5 were unable to appreciably enhance the weak visual effect relative to the strong auditory effect (from Experiment 3). Overall, we found that both auditory and visual cues reliably conveyed statement and question intonation, were successfully synthesized, and generalized to other utterances. However, the weak visual effect relative to the robustly strong auditory effect precluded optimal integration and conclusive examination of information processing through model fitting. PMID- 14529111 TI - Effects of pitch accent position, type, and status on focus projection. AB - This paper examines predictions made by two theories of the relationship between pitch accent and focus. The empirical evidence presented suggests that listeners are sensitive to a variety of factors that may affect the focus projection ability of pitch accents, that is the ability of a pitch accent on one word to mark focus on a larger constituent. The findings suggest that listeners' interpretation of focus structure is most sensitive to the presence or absence of a pitch accent on a focused constituent and the deaccenting of following unfocused material (pitch accent position). Preliminary evidence suggests that the status of a pitch accent as nuclear or prenuclear may also affect listeners' interpretations, though to a lesser extent than accent position. Finally, the results show that focus projection is affected only minimally, if at all, by the type of pitch accent (at least for the two accent types compared (H* vs. L + H*)). PMID- 14529110 TI - Reanalysis of clause boundaries in Japanese as a constraint-driven process. AB - When processing the initial segment of a sentence, readers may favor an interpretation that will turn out to be incorrect as more words are read. In these cases, a reanalysis process is necessary in order to correct the mental representation built up to that point. It has been previously proposed that readers obey a minimum change restriction, as they prefer to change the mental representation as little as possible. The present paper reports two experiments in Japanese suggesting that a minimal change restriction is unnecessary to characterize reanalysis. It is proposed instead that the present data and previous observations are more naturally explained by a constraint-driven model in which revisions are performed only when required by parsing constraints. PMID- 14529112 TI - Who is my neighbour? PMID- 14529113 TI - Undertaking the role of patient advocate: a longitudinal study of nursing students. AB - Patient advocacy has been claimed as a new role for professional nurses and many codes of ethics for nurses state that they act as patient advocates. Nursing education is faced with the challenge of preparing nurses for this role. In this article we describe the results of a study that considered the tendencies of a cohort of nursing students at the Kocaeli University School of Nursing to act as advocates and to respect patients' rights, and how their capacities to do so changed (or not) as a result of their nursing education. This longitudinal study used a questionnaire consisting of 10 statements relating to patient care. It was performed both at the start (1998) and at the end (2002) of the nursing training. At the beginning of their course 77 students participated; in the study. After four years, only 55 students participated, the reason for this drop in number being unknown. The questions asked nurses if patients should have: the right to receive health care; the right to participate in the decision-making process about their treatment; the right always to be told the truth; and the right to have access to their own medical records. They were also asked: if quality of life should be a criterion for discontinuing treatment; if patients have the right to die and the right to refuse treatment; if patients should be assisted to die or helped to undergo active euthanasia; and if severely disabled newborn babies should be allowed to die. The student nurses demonstrated considerable insight into contemporary nursing issues and were ready to act as patient advocates. Professional responsibility demands that good nurses advocate strongly for patients' choices. PMID- 14529114 TI - A study of the ethical sensitivity of physicians in Turkey. AB - In order to prepare bioethics and clinical ethics courses for clinicians in Turkey, we needed to know the attitudes of physicians when placed in ethically difficult care situations. We presented four cases to 207 physicians who are members of the Physicians' Association in Kocaeli, Turkey. Depending on the decisions they made in each case, we determined whether they were aware of the ethical aspects of the cases and the principles they chose as a basis for their decisions. We aimed to gain information about their ethical tendencies and moral sensitivity. A small number of physicians stated that they would 'show respect for a patient's living will' in the first case, but more stated that they would 'let the patient refuse the treatment' in the second. In the third case, where medical confidentiality was the significant ethical issue, most of the physicians said that they would act in order to maintain confidentiality. For the last case, more than half the physicians chose to 'tell the truth' to the patient. The paternalism shown in the doctors' decisions on the first two cases was no longer observed in those made for the last two cases. We concluded that the physicians who participated in our study have low sensitivity to living wills (or advance directives) and patients refusing treatment. However, when issues of medical confidentiality and truth-telling are concerned, they take care to protect the autonomy of the individual an are relatively more aware of the ethical aspects of these cases. PMID- 14529115 TI - Nurses' and midwives' views on approaches to hymen examination. AB - Premarital sexual relations are unacceptable for women within Turkish society's understanding and perception of honour. If there is any suspicion about virginity, young girls are forced to undergo hymen examination against their will, which frequently results in attemped suicide. The most frequent cause of suicide in young Turkish girls is hymen examination. Nurses and midwives are always involve in this procedure. The purpose of this study was to determine the views of and approaches to hymen examination by nurses and midwives. Of those who participated in this study, 80.2% had been present during a hymen examination, 40.4% indicated that virginity had a special significance in the society owing to social pressures, and 37.5% indicated that they agreed with the view that the idea of virginity puts restraints on a woman's sexual life, whereas sexual activity is a physiological need. They also indicated their opposition to hymen examination imposed without the individual's consent. They agreed that stopping this practice depended more on the social structural changes required than on legal measures. PMID- 14529116 TI - The death of a patient with AIDS in Turkey: thoughts on the ethical dimensions. AB - A Turkish patient with AIDS attempted to commit suicide. Turkey is one of the countries where AIDS education in society and for health personnel has started rather late. This article documents what this patient, his sister and his friends, who helped him to survive for a short while, experienced in the hospital environment. This is a real case history and should be considered from various aspects because suicide was attempted by a person with AIDS who was near the terminal stage. The patient's friends made a remarkable effort to help him to survive. They were unsure about what to do because they had not respected his wishes and they experienced deep alienation because of the extremely negative attitude of the medical staff. PMID- 14529117 TI - Ethical issues in adolescent consent for research. AB - Different opinions are expressed in the literature regarding when children and adolescents can start to make decisions to participate in research and give informed consent. Nurses are frequently involved in research, either as investigators or caregivers, and must therefore have a thorough understanding of consent and related issues. In this article the issues are explored from a Canadian perspective. The argument is put forward that adolescents may be capable of a greater involvement in the research consent process than is the norm. Increasing adolescents' involvement in research has the potential to enhance their growing autonomy and capabilities. Adolescents appreciate being treated with respect and dignity by adults. This can be achieved in an environment in which protection from harm does not also mean prevention from decision making. The use of empowering processes by nurses to enhance adolescent involvement will provide benefit to adolescents in their transition to adult levels of responsibility. PMID- 14529118 TI - The use of physical restraints for patients suffering from dementia. AB - This study reviews the ethical dilemmas of nursing staff about using restraints on patients suffering from dementia in two types of health care settings in Israel: internal medicine wards of three general hospitals; and psychogeriatric wards of three nursing homes. The nurses' level of knowledge about the Patient's Rights Law, the Israeli Code of Ethics, and the guidelines on restraints was analysed. The purposes of restraints were defined as beneficial to: (1) the patient; (2) other patients; or (3) the institution. The concept was evaluated in a realistic situation (expressing views of daily practice) and in an idealistic situation (expressing personal and professional beliefs and values). It was shown that nurses in internal medicine wards of general hospitals agreed more with the use of restraints than those in psychogeriatric wards in nursing homes. Differences were more pronounced when restraints were beneficial to the institution. In addition, nurses working in psychogeriatric wards of nursing homes had more knowledge about the guidelines on restraints and were less inclined than their counterparts to agree with the use of restraints for the benefit of other patients or the institution. PMID- 14529120 TI - Nurses' ethical conflicts in performance of utilization reviews. AB - This article describes the ethical conflicts that a sample of US nurse utilization reviewers faced in their work, and also each nurse's self-reported ethical orientation that was used to resolve the dilemmas. Data were collected from a sample of 97 registered nurses who were working at least 20 hours per week as utilization reviewers. Respondents were recruited from three managed care organizations that conduct utilization reviews in a large midwestern city. A cross-sectional survey design was used to collect demographic data and to ask closed-response, short-answer and open-ended questions. Ethical conflicts reported by nurses were similar across utilization review settings and many were justice orientated. Self-reported ethical orientations were similar across organizations, with beneficence dominating. Implications of these findings are discussed. PMID- 14529121 TI - An interview with Saadet Ulker. PMID- 14529119 TI - Whistleblowing and boundary violations: exposing a colleague in the forensic milieu. AB - The purpose of this article is to examine the phenomenon of whistleblowing as it relates to a reconstructed case study of an erotic boundary violation that emerged from a clinical situation in forensic psychiatric nursing practice. The unique features of this case are illustrated with the help of a model for decision making. Although the ramifications of exposing a colleague are many, it is argued that, in this particular case, it was morally and ethically the right thing to do. PMID- 14529122 TI - Is ethics necessary in medicine? PMID- 14529123 TI - Latest developments in medical ethics in Turkey. PMID- 14529124 TI - Urine sulfated and nonsulfated bile acids as a diagnostic test for liver disease in cats. PMID- 14529125 TI - The oomycetes--fungi with teeth and flippers? PMID- 14529126 TI - A review of methods for assessment of the rate of gastric emptying in the dog and cat: 1898-2002. AB - Gastric emptying is the process by which food is delivered to the small intestine at a rate and in a form that optimizes intestinal absorption of nutrients. The rate of gastric emptying is subject to alteration by physiological, pharmacological, and pathological conditions. Gastric emptying of solids is of greater clinical significance because disordered gastric emptying rarely is detectable in the liquid phase. Imaging techniques have the disadvantage of requiring restraint of the animal and access to expensive equipment. Radiographic methods require administration of test meals that are not similar to food. Scintigraphy is the gold standard method for assessment of gastric emptying but requires administration of a radioisotope. Magnetic resonance imaging has not yet been applied for assessment of gastric emptying in small animals. Ultrasonography is a potentially useful, but subjective, method for assessment of gastric emptying in dogs. Gastric tracer methods require insertion of gastric or intestinal cannulae and are rarely applied outside of the research laboratory. The paracetamol absorption test has been applied for assessment of liquid phase gastric emptying in the dog, but requires IV cannulation. The gastric emptying breath test is a noninvasive method for assessment of gastric emptying that has been applied in dogs and cats. This method can be carried out away from the veterinary hospital, but the effects of physiological and pathological abnormalities on the test are not known. Advances in technology will facilitate the development of reliable methods for assessment of gastric emptying in small animals. PMID- 14529127 TI - Persistent urinary tract infections and reinfections in 100 dogs (1989-1999). AB - A retrospective study was performed of 100 dogs with persistent urinary tract infections (UTIs) or reinfections presenting to the North Carolina State University (Raleigh, NC) Veterinary Teaching Hospital between 1989 and 1999. Criteria for selection included > or = 2 positive urine cultures within a 6-month period. Signalment, presence of predisposing disorders, urinalysis and urine culture results, and treatment strategies were extracted from the medical records. Dogs were a median age of 7 years when the UTI was 1st diagnosed. Dogs younger than 3 and older than 10 years were at increased and decreased risks, respectively, for reinfections or persistent UTIs. Spayed females were more common in the UTI population. More than half of the dogs were asymptomatic for a UTI at 1st presentation. Urine sediment examinations identified hematuria, pyuria, and bacteriuria in 47, 72, and 85% of the samples, respectively. The most commonly isolated organisms were Escherichia coli and Streptococcus/Enterococcus spp.; multiple isolates also were common. Of the isolates, 29.5% were resistant to achievable serum concentrations of all antibiotics commonly prescribed for PO administration. Dogs with abnormal micturition were more likely to have infections by organisms resistant to commonly prescribed antibiotics. Potentially predisposing disorders were identified in 71 dogs. A correction of these disorders was accomplished in 35% of these 71 dogs. Dogs given standard antibiotic therapy without addressing predisposing disorders experienced poor control of their UTIs; 74.5% of these dogs had an apparent disease-free interval (ADFI) of < 8 weeks. By comparison, dogs in which predisposing disorders were corrected or those that were treated with low-dose, long-term antibiotic regimens subjectively had better control. PMID- 14529128 TI - Intermittent bolus injection versus continuous infusion of furosemide in normal adult greyhound dogs. AB - Several studies in human subjects have demonstrated greater diuresis with constant rate infusion (CRI) furosemide than intermittent bolus (IB) furosemide. This study was conducted to compare the diuretic efficacy of the same total dose of IB furosemide and CRI furosemide in 6 healthy, adult Greyhound dogs in a randomized crossover design with a 2-week washout period between treatments. For IB administration, dogs received 3 mg/kg at 0 and 4 hours. For CRI administration, dogs received a 0.66 mg/kg loading dose followed by 0.66 mg/kg/h over 8 hours. The same volume of fluid was administered for both methods. Urine output was quantified hourly. Urine electrolyte concentrations, urine specific gravity (USG), packed cell volume (PCV), total protein (TP), serum electrolyte concentrations, total carbon dioxide (TCO2), serum creatinine (sCr), and blood urea nitrogen (BUN) were determined every 2 hours. Urine production and water intake were greater (P < or = 0.05) for CRI than IB. Urine sodium and calcium losses were greater (P < 0.05) and urine potassium loss was less (P = 0.03) for CRI than IB, but there was no evidence of a difference between methods for urine magnesium and chloride losses. Serum chloride concentration was less (P < 0.001), sCr concentration greater (P = 0.04). TP greater (P = 0.01), and PCV greater (P = 0.003) for CRI than IB. No differences in USG, TCO2, BUN, or serum potassium, sodium, and magnesium concentrations were detected between methods. The same total dose of CRI furosemide resulted in more diuresis, natriuresis, and calciuresis and less kaliuresis than IB furosemide in these normal Greyhound dogs over 8 hours, suggesting that furosemide is a more effective diuretic when administered by CRI than by IB. PMID- 14529129 TI - Clinicopathologic findings associated with Lagenidium sp. infection in 6 dogs: initial description of an emerging oomycosis. AB - An oomycotic pathogen in the genus Lagenidium was isolated from tissues obtained from 6 dogs with progressive cutaneous disease. Initial clinical findings in 5 dogs included multifocal cutaneous lesions, subcutaneous lesions, or both associated with regional lymphadenopathy: the 6th dog initially was presented for evaluation of mandibular lymphadenopathy. Cutaneous lesions were ulcerated, exudative regions (often with necrosis and draining tracts) or multiple firm dermal or subcutaneous nodules. Two dogs subsequently developed hemoabdomen from great vessel rupture and died acutely. Four dogs were euthanized because of progression of subcutaneous lesions or lymphadenopathy. On postmortem examination, regional granulomatous lymphadenitis was found in all 6 dogs, great vessel invasion in 3 dogs, pulmonary lesions in 2 dogs. ureteral obstruction in 1 dog, mediastinal lymphadenitis in 1 dog, and hilar lymphadenitis with invasion of the distal esophagus and trachea in 1 dog. Histologically, lesions were similar to those associated with pythiosis and zygomycosis and were characterized by severe eosinophilic granulomatous inflammation (often with numerous large multinucleated giant cells) centered around broad (7-25 micro), infrequently septate hyphae. Immunoblot analysis of the serologic response of 4 dogs to a soluble mycelial extract of Lagenidium giganteum indicated that each dog's serum recognized at least 10 different antigens of L. giganteum. Culture of infected tissues yielded rapid growth of colorless to white submerged colonies. Microscopically, mature hyphae in culture were broad (25-40 micro), segmented, and occasionally branching and produced motile laterally biflagellate zoospores in water culture. This report is the 1st description of infection caused by an oomycete other than Pythium insidiosum in any mammalian species. PMID- 14529130 TI - Clinical findings in 40 dogs with hypersensitivity associated with administration of potentiated sulfonamides. AB - The purpose of this study was to summarize the clinical findings in 40 dogs with systemic hypersensitivity reactions associated with the administration of potentiated sulfonamides. Dogs ranged from 6 months to 14 years of age, with a mean of 5.7 +/- 3.2 years. Spayed female dogs were overrepresented (24 of 40, or 60% of the dogs), as were Samoyeds (3 of 40; 8%) and Miniature Schnauzers (5 of 40; 13%). Mean dosages of potentiated sulfonamides were 47.0 +/- 14.9 mg/kg/d (range, 23.4-81.4 mg/kg/d). The time from the 1st administration of the drug to the onset of the clinical signs of hypersensitivity ranged from 5 to 36 days, with a mean of 12.1 +/- 5.9 days. There was no relationship between either the dosage or type of sulfonamide given and the time to the onset of the clinical signs. Fever was the most common clinical sign observed (55% of the dogs); thrombocytopenia was 2nd (54%), and hepatopathy (28%) was 3rd. Neutropenia, keratoconjunctivitis sicca (KCS), hemolytic anemia. arthropathy, uveitis, skin and mucocutaneous lesions, proteinuria, facial palsy, suspected meningitis, hypothyroidism, pancreatitis, facial edema, and pneumonitis were also observed in some patients. Of 39 dogs with adequate follow-up, 30 (77%) recovered, whereas 8 (21%) either died or were euthanized, and 1 recovered clinically but had persistent increases in alanine aminotransferase (ALT) activity. Dogs with hepatopathy generally had a poorer prognosis (46% recovery) than dogs without hepatopathy (89% recovery; P = .0035). Sixty-three percent of the dogs with thrombocytopenia recovered, compared to 90% of the dogs without thrombocytopenia (P = .042). Recovery was not associated with sex, age, breed, or type of sulfonamide administered. PMID- 14529131 TI - M-mode echocardiographic ratio indices in normal dogs, cats, and horses: a novel quantitative method. AB - A novel method for quantitative echocardiographic interpretations is introduced based on the calculation of ratio indices in which each raw M-mode measurement is divided by the aortic root dimension (Ao). "Aorta-based" indices were calculated with the animal's measured aortic root dimension (Ao(m)) as the length standard. Conversely, "weight-based" indices employed an idealized estimate of aortic dimension (Ao(w)) with a weighted least squares linear regression against the cube root of body weight (Ao(w) = kW(1/3)). Use of these indices circumvented undesirable statistical characteristics inherent in linear regression of echocardiographic dimensions against body weight and, to a lesser extent, body surface area. Compared with the regressions, ratio indices resulted in substantial refinement of the predictive range for each M-mode measurement in dogs, particularly with decreasing body size. Weight-based indices outperformed aorta-based indices in this regard. To refine the predictive range, neither type of index was clearly advantageous in cats compared with the simple average method typically employed for that species. Several of the raw M-mode measurements, however, were correlated with body weight in cats and horses, indicating the need for an appropriate correction for body size in these species. The ratio index method was suitable for this purpose. Summary statistics derived from normal dogs (n = 53), cats (n = 32), and horses (n = 17) are presented for each index, including novel clinical indices calculated from area ratios. The latter were designed to represent body size-adjusted lett ventricular stroke area (ie, volume overload) and myocardial wall area (ie, hypertrophy). PMID- 14529132 TI - Noninvasive transthoracic temporary cardiac pacing in dogs. AB - Temporary cardiac pacing is used in the emergency treatment of life-threatening bradyarrhythmias and for the support of heart rate and blood pressure of patients with sick sinus syndrome or high-grade atrioventricular (AV) block undergoing general anesthesia, typically for permanent pacemaker implantation. We retrospectively evaluated the safety and efficacy of a noninvasive transthoracic external cardiac pacing system in 42 dogs treated for bradyarrhythmias. Optimal placement of the patch electrodes on the skin of the thorax was initially established on 2 anesthetized normal dogs. The optimal electrode placement was determined to be on the right and left hemithoraces, directly over the heart. Afterward, by means of this electrode placement all 42 dogs treated for bradyarrhythmias in this study were successfully paced with the noninvasive transthoracic system. Dogs ranged in age from 1 to 15 years and weighed between 3.2 and 40 kg. Miniature Schnauzers, German Shepherds, and mixed breeds were most common in the study population. Indications for noninvasive transthoracic pacing included emergency treatment of hemodynamically unstable 3rd-degree AV block (2 dogs): support of heart rate during general anesthesia for permanent pacemaker implantation or lead-wire adjustment (38 dogs): and support of heart rate during general anesthesia for ophthalmologic surgery in dogs with sick sinus syndrome (2 dogs). Complications included pain and skeletal muscle stimulation, which required general anesthesia. We conclude that the noninvasive transthoracic pacing system evaluated is satisfactory for clinical veterinary use. PMID- 14529133 TI - Cisplatin and doxorubicin toxicosis in dogs with osteosarcoma. AB - Toxicosis associated with doxorubicin and cisplatin administration starting either 2 or 10 days after limb amputation for osteosarcoma was examined retrospectively in dogs. The purpose was to determine whether dosage and timing of chemotherapy affected rates of toxicosis after administration of the 1st treatment. Records of 100 dogs with appendicular osteosarcoma without evidence of metastases or concurrent disease were examined. Dogs received chemotherapy with doxorubicin and cisplatin every 3 weeks for 3 treatments starting 2 days (n = 51) or 10 days (n = 49) after amputation. The dosage of cisplatin was 60 mg/m2 and was given with 6-hour saline diuresis and butorphanol. Doxorubicin was given at 12.5-25 mg/ml during fluid administration. Hematologic data were collected before and weekly after treatment. Client interviews were conducted to assess gastrointestinal toxicosis during the interval between treatments. The reported toxicoses were graded on a scale of 0 to 4. Dogs receiving 25 mg/m2 of doxorubicin experienced greater rates of grade 4 toxicity (67%; n = 6) than dogs in groups receiving 12.5-20 mg/m2 of doxorubicin (< or = 25%; n = 94, P = .03). Dogs in the Day 2 group experienced greater rates (35%) of grade 4 toxicity than dogs in the Day 10 group (12%, P = .007). We concluded that chemotherapy administered 2 days after surgery produced an unacceptable level of toxicoses. except at greatly reduced dosages, and that even with a delay of treatment, 25 mg/m2 of doxorubicin, when given in combination with cisplatin at 60 mg/m2, was too toxic for routine use. PMID- 14529134 TI - Hemostatic changes in dogs with naturally occurring sepsis. AB - Sepsis is a frequent source of morbidity and mortality in critically ill patients. The goal of this case control study was to measure hemostatic changes in dogs with naturally occurring sepsis. Blood was collected within 24 hours of admission from 20 dogs that fulfilled the criteria for sepsis. Sepsis was defined as histologic or microbiological confirmation of infection and 2 or more of the following criteria: hypo- or hyperthermia, tachycardia, tachypnea, or leukopenia, leukocytosis, or > 3% bands. Culture and sensitivities were performed on appropriate samples from all septic dogs. Twenty-eight control dogs were enrolled on the basis of normal results of physical examination, CBC, serum biochemistry, and coagulation profile. Plasma samples were analyzed for prothrombin time (PT), partial thromboplastin time (PTT), fibrin(ogen) degradation products (FDP), D dimer (DD) concentrations, antithrombin (AT) activity, and protein C (PC) activity. Data were compared between groups by chi-square or independent t-tests. PC (P < .001) and AT (P < .001) activities were significantly lower in dogs with sepsis compared to controls. Dogs with sepsis had significantly higher PT (P = .007), PTT (P = .005), D-dimer (P = .005), and FDP (P = .001) compared to controls. Platelet counts were not significantly different between groups. Ten of the 20 septic dogs (50%) died, but no association was identified between any of the measured variables and outcome. These findings are consistent with previous studies in animals with experimentally induced disease and in clinical studies of humans. On the basis of these results, further investigation of the role of AT and PC in canine sepsis is warranted. PMID- 14529135 TI - Decreased platelet function in Cavalier King Charles Spaniels with mitral valve regurgitation. AB - With aggregometry, increased platelet activity has been reported in Cavalier King Charles Spaniels (CKCS) without mitral regurgitation (MR). In contrast, dogs with MR have been found to have decreased platelet activity. The purpose of this study was to test an easy bedside test of platelet function (the Platelet Function Analyzer [PFA-100]) to see if it could detect an increase in platelet activity in CKCS without MR and a decrease in platelet activity in CKCS with MR. This study included 101 clinically healthy dogs > 1 year of age: 15 control dogs of different breeds and 86 CKCS. None of the dogs received medication or had a history of bleeding. The PFA-100 evaluates platelet function in anticoagulated whole blood under high shear stress. Results are given as closure times (CT): the time it takes before a platelet plug occludes a hole in a membrane coated by agonists. The CT with collagen and adenosine-diphosphate as agonists was similar in control dogs (median 62 seconds; interquartile interval 55-66 seconds) and CKCS with no or minimal MR (55; 52-64 seconds). The CT was higher in CKCS with mild MR (regurgitant jet occupying 15-50% of the left atrial area) (75; 60-84 seconds; P = .0007) and in CKCS with moderate to severe MR (jet > 50%) (87: 66 102 seconds; P < .0001). CKCS with mild, moderate, and severe, clinically inapparent MR have decreased platelet function. The previous finding of increased platelet reactivity in nonthrombocytopenic CKCS without MR could not be reproduced with the PFA-100 device. PMID- 14529136 TI - Biologic behavior and prognostic factors for mast cell tumors of the canine muzzle: 24 cases (1990-2001). AB - The medical records of 24 dogs with histologically confirmed mast cell tumors (MCT) of the muzzle were retrospectively evaluated to determine their biologic behavior and prognostic factors. Information on signalment, tumor grade and stage, treatment methods, and pattern of and time to failure and death was obtained from the medical record. Twenty-three dogs were treated with combinations of radiotherapy, surgery, and chemotherapy; 1 dog received no treatment. There were 2 Grade 1, 15 Grade 11, and 7 Grade III tumors. Tumors were stage 0 (n = 8), stage 1 (5), stage 2 (6), stage 3 (4), and stage 4 (1). Mean and median survival times of treated dogs were 36 and 30 months, respectively. Prognostic factors affecting survival time included tumor grade and presence of metastasis at diagnosis. Dogs with Grade I and II tumors survived longer than dogs with Grade III tumors. Variables, including sex, age, gross versus microscopic disease, and treatment type were not found to affect survival. Local control rate was 75% at 1 year and 50% at 3 years. Tumor grade was the only variable found to affect local control. Dogs with Grade I tumors had longer disease-free intervals than those with Grade II tumors, and dogs with Grade II tumors had longer disease-free intervals than dogs with Grade III tumors. Eight of 9 dogs dying of MCT had local or regional disease progression. Muzzle MCT a rebiologically aggressive tumors with higher regional metastatic rates than previously reported for MCT in other sites. PMID- 14529137 TI - Effect of dietary starch, fat, and bicarbonate content on exercise responses and serum creatine kinase activity in equine recurrent exertional rhabdomyolysis. AB - To determine the effect of dietary starch, bicarbonate, and fat content on metabolic responses and serum creatine kinase (CK) activity in exercising Thoroughbreds with recurrent exertional rhabdomyolysis (RER), 5 RER horses were fed 3 isocaloric diets (28.8 Mcal/d [120.5 MJ/d]) for 3 weeks in a crossover design and exercised for 30 minutes on a treadmill 5 days/wk. On the last day of each diet, an incremental standardized exercise test (SET) was performed. The starch diet contained 40% digestible energy (DE) as starch and 5% as fat: the bicarbonate-starch diet was identical but was supplemented with sodium bicarbonate (4.2% of the pellet): and the fat diet provided 7% DE as starch and 20% as fat. Serum CK activity before the SET was similar among the diets. Serum CK activity (log transformed) after submaximal exercise differed dramatically among the diets and was greatest on the bicarbonate-starch diet (6.51 +/- 1.5) and lowest on the fat diet (5.71 +/- 0.6). Appreciable differences were observed in the severity of RER among individual horses. Postexercise plasma pH, bicarbonate concentration, and lactate concentration did not differ among the diets. Resting heart rates before the SET were markedly lower on the fat diet than on the starch diet. Muscle lactate and glycogen concentrations before and after the SET did not differ markedly among the diets. A high-fat, low-starch diet results in dramatically lower postexercise CK activity in severely affected RER horses than does a low-fat, high-starch diet without measurably altering muscle lactate and glycogen concentrations. Dietary bicarbonate supplementation at the concentration administered in this study did not prevent increased serum CK activity on a high-starch diet. PMID- 14529138 TI - Botulism in foals less than 6 months of age: 30 cases (1989-2002). AB - Botulism has been recognized as a clinical entity in foals since the 1960s. Also known as "Shaker foal" disease, the toxicoinfectious form of botulism affects foals, with the highest incidence in the United States seen in Kentucky and the mid-Atlantic region. The disease is characterized by progressive muscular weakness caused by the action of botulism neurotoxin at cholinergic neuromuscular junctions. Increased number of episodes and duration of recumbency, muscular trembling, and dysphagia are seen in affected foals. Left untreated, the disease can be rapidly fatal, with death occuring secondary to respiratory muscle paralysis within 24 to 72 hours of the onset of clinical signs. Very mildly affected foals can survive with minimal treatment Despite advances made in treatment of these foals, including administration of botulism antitoxin early in the course of the disease, there is still an impression that the disease carries a high mortality rate. The purpose of this study was to evaluate outcome in 30 foals <6 months of age diagnosed with botulism between 1989 and 2002 at the George D. Widener Large Animal Hospital, New Bolton Center. Two foals were euthanized for economic reasons early in the disease course, and I died while being treated. Survival of treated cases was greater than 96%. Approximately 50% of the cases required oxygen therapy, whereas 30% required mechanical ventilation. All foals, excepting 1 mildly affected foal, received botulism antitoxin. Mean duration of hospitalization was 14 days. With appropriate treatment, foals with botulism have a high survival rate. PMID- 14529139 TI - Mechanical ventilation in foals with botulism: 9 cases (1989-2002). AB - "Shaker foal" disease, toxicoinfectious botulism of foals, was 1st described as a clinical entity in 1967. The reported mortality rate was 90%, with death occurring within 24-72 hours of the onset of the characteristic clinical signs. The mortality rate decreased when equine-origin botulism antitoxin became available; however, a certain percentage of foals continued to die of respiratory failure. Mechanical ventilation is an important part of the treatment of infant botulism and is essential to the survival of many affected infants. We report a retrospective study of 9 foals with toxicoinfectious botulism where early mechanical ventilation was employed as part of the treatment. Foals receiving mechanical ventilation were progressively acidemic and had increased PaCO2 tensions before mechanical ventilation. These arterial blood gas abnormalities were ameliorated with mechanical ventilation. One foal was euthanized for economic reasons; survival in treated foals was 87.5%. Mechanical ventilation of foals with botulism and respiratory failure appears to be an effective therapy. PMID- 14529140 TI - First pacemaker in a dog: a historical note. PMID- 14529141 TI - Transvenous electrical cardioversion in equine atrial fibrillation: technique and successful treatment of 3 horses. PMID- 14529142 TI - Thiamine deficiency in a dog: clinical, clinicopathologic, and magnetic resonance imaging findings. PMID- 14529143 TI - Short-term compatibility of furosemide with crystalloid solutions. AB - Parenteral veterinary furosemide is a 50-mg/mL solution with a pH of 8.0-9.3. The purpose of this study was to determine whether a commonly used veterinary formulation of 50 mg/mL of furosemide solution could be diluted in vitro without precipitation. Furosemide 50 mg/mL was diluted to concentrations of 10 and 5 mg/mL with 5% dextrose in water (D5W), 0.9% saline, lactated Ringer solution (LRS), and sterile water. Acidic sterile water and basic sterile water solutions were made by the addition of hydrochloric acid and sodium hydroxide, respectively, for use as controls to assess the effect of pH extremes for each concentration. After furosemide dilution, the final pH of each sample was measured, and samples were grossly and microscopically examined for clarity and crystal formation immediately and 1, 3, 5, and 8 hours after dilution. Gross precipitation and microscopic crystals were immediately observed in the acidic controls. Solutions of 5 mg/mL in LRS and 0.9% saline became slightly cloudy immediately, but no crystals were observed microscopically for 8 hours. Solutions of 10 mg/mL in D5W, 0.9% saline, LRS, and sterile water and solutions of 5 mg/mL in D5W and sterile water and the basic control were grossly clear, and no microscopic crystals were observed for 8 hours. On the basis of the results obtained in this in vitro investigation, this veterinary formulation of furosemide 50 mg/mL can be diluted without precipitation to a concentration of 10 mg/mL with D5W, 0.9% saline, LRS, or sterile water and to 5 mg/mL with D5W or sterile water and held for 8 hours. PMID- 14529144 TI - Photodynamic therapy for the treatment of intranasal tumors in 3 dogs and 1 cat. AB - Three dogs and 1 cat with intranasal tumors were treated with pyropheophorbide-a hexyl ether-based photodynamic therapy (PDT). PDT was well tolerated by all the animals, and no adverse effects from photosensitizer injection, such as cutaneous photosensitization, were observed. Facial swelling was observed in all animals after each PDT treatment but resolved spontaneously within 72 hours after treatment. All animals had a decrease in severity of epistaxis, frequency of sneezing, and amount of nasal discharge after PDT. Clinical signs were controlled for variable time, although long-term responses were comparable with radiation therapy in 2 animals. This small case series demonstrates another application for PDT in veterinary medicine. On the basis of these findings. further studies are warranted to define the role of PDT in the management of intranasal tumors in dogs and cats. PMID- 14529145 TI - Myelopathy in Holstein x Gir calves in Brazil. AB - This case report contains clinical and pathologic features of a degenerative myelopathy in Holstein X Gir crossbred calves in Brazil. The bilateral and symmetrical spinal cord white matter lesions were interpreted as a primary axonopathy that may be of the dying-back type. PMID- 14529146 TI - Treatment with vitamin D3 and/or vitamin K2 for postmenopausal osteoporosis. AB - It is established in Japan that treatment with 1alpha-hydroxyvitamin D3 (alfacalcidol) slightly reduces bone turnover, sustains lumbar bone mineral density (BMD), and prevents osteoporotic vertebral fractures in postmenopausal women with osteoporosis, while vitamin K2 (menatetrenone) enhances gamma carboxylation of bone glutamic acid residues and secretion of osteocalcin, sustains lumbar BMD, and prevents osteoporotic fractures in patients with osteoporosis. Available evidence suggests that the effect of vitamin K2 on mineralization by human periosteal osteoblasts is enhanced in the presence of 1,25 dihydroxyvitamin D3 in vitro. The effect of vitamin K2 on BMD in ovariectomized rats is affected by the plasma 25-hydroxyvitamin D3 level in vivo, and is significant only when rats are fed a diet containing vitamin D3. Based on this line of evidence, combined treatment with alfacalcidol and menatetrenone for osteoporosis is surmised to be more effective than treatment with menatetrenone alone, and may have anabolic effects on osteoporotic bone. This combined treatment may increase bone formation as well as bone resorption over the mild anti-resorptive effect of alfacalcidol itself, and shows the greatest effect on lumbar BMD or the incidence of vertebral fractures in studies in which the mean age and years since menopause of subjects were low and the degree of osteoporosis was mild. It may be effective for mild postmenopausal osteoporosis in which age related deterioration of trabecular bone properties remains below the threshold for vertebral fractures, even if bone resorption is increased and trabecular bone has deteriorated. PMID- 14529147 TI - Cell choice for bioartificial livers. AB - It is unlikely that human hepatocytes can be isolated on a scale sufficient to treat more than a fraction of the patients who need bioartificial liver (BAL) treatment. The use of animal cells results in the concerns related to the transmission of infectious pathogens and immunologic and physiologic incompatibilities between the donor and humans. Human embryonic stem cells and bone marrow multipotent adult progenitor cells have received great attention as a possible source for BALs. The use of tightly regulated clonal hepatocyte cell lines would be attractive. Such cell lines grow economically in tissue culture and provide the advantage of uniformity, sterility, and freedom of pathogens. In this paper, the authors review the choice of cells for BALs and discuss our reversible immortalization system of human liver cells using a retroviral transfer of immortalizing genes and subsequent Cre/loxP-mediated site-specific recombination. PMID- 14529148 TI - Gingipains as candidate antigens for Porphyromonas gingivalis vaccine. AB - Porphyromonas gingivalis (P. gingivalis), a gram-negative anaerobe, is involved in the pathogenesis of periodontal disease, and is found frequently in the subgingival flora in patients with periodontitis. This organism possesses a variety of virulence factors including lipopolysaccharide, capsular material, fimbriae and proteases (enzymes). Among the P. gingivalis antigens, enzymes such as Arginine-specific gingipains (RgpA, RgpB) and lysine-specific gingipain (Kgp) have been studied for their ability to induce biologically significant antibodies. This review summarizes recent information on the gingipains and their possible application in the development of an anti-P. gingivalis vaccine. PMID- 14529149 TI - Current consensus on the diagnosis and treatment of H. pylori-associated gastroduodenal disease. AB - Helicobacter pylori (H. pylori) is a spiral shaped bacterium that resides in the stomach mucosa. Isolation of H. pylori from the stomach mucosa changed the erstwhile widely held belief that the stomach contains no bacteria and is actually sterile. Once H. pylori is safely ensconced in the mucus, it is able to neutralize the acid in the stomach by elaborating an enzyme called urease. Urease converts urea, of which there is an abundant supply in the stomach (derived from saliva and the gastric juice), into bicarbonate and ammonia, which are strong bases. These bases form a cloud of acid-neutralizing chemicals in the vicinity of the organisms, protecting them from the acid in the stomach. This urea hydrolysis reaction is utilized for the diagnosis of H. pylori infection in the urea breath test (UBT) and the rapid urease test (RUT). In Japan, both invasive tests, such as bacterial culture, histopathology and RUT, and non-invasive tests such as UBT and serology are conducted for the diagnosis of H. pylori infection. For confirming the results of eradication therapy, UBT is considered to be the most sensitive and specific. In order to treat H. pylori infection, a new one-week triple therapy regimen (lansoprazole or omeprazole + amoxicillin + clarithromycin) has been approved for use in patients with peptic ulcer disease in Japan. As for H. pylori eradication in the case of other diseases in which the bacterium has been implicated (e.g., chronic atrophic gastritis, gastric MALT lymphoma, gastric cancer, non-ulcer dyspepsia, chronic urticaria, idiopathic thrombocytopenic purpura (ITP)), further basic and clinical investigation is required. PMID- 14529150 TI - Molecular mechanism of chicken ovalbumin upstream promoter-transcription factor (COUP-TF) actions. AB - Chicken ovalbumin upstream promoter-transcription factors (COUP-TFs) are one of the most characterized orphan receptors of the steroid/thyroid hormone receptor superfamily. COUP-TFs play important roles in the regulation of organogenesis, neurogenesis, and cellular differentiation during embryonic development. COUP-TFs were generally considered to be repressors of transcription, however, there are growing evidences that COUP-TFs can function as transcription activators. Here we will review the molecular mechanism of COUP-TFs as repressors and activators. Also, we will review the known biological function of COUP-TFI during development and differentiation. PMID- 14529151 TI - Current results on the use of imatinib mesylate in patients with relapsed Philadelphia chromosome positive leukemia after allogeneic or syngeneic hematopoietic stem cell transplantation. AB - Here, we describe a patient diagnosed with chronic myelogenous leukemia who relapsed after matched unrelated donor SCT. The patient was treated with imatinib mesylate and donor lymphocyte infusions, and achieved a complete molecular remission. Additionally, safety and efficacy of imatinib mesylate in a total of 134 patients from 8 centers who underwent allogeneic or syngeneic stem cell transplantation (SCT) and had a relapse of Philadelphia chromosome positive leukemia was reviewed. Data was compiled from abstracts accepted as oral or poster presentations at the ASH (American Society of Hematology) 2001 and EBMT (European Group for Blood and Marrow Transplantation) 2001 & 2002 meetings and additionally literature published on this patient group. Efficacy of imatinib therapy was assessed by morphology, cytogenetic analysis, and determination of donor chimerism. In the evaluable population, hematologic and cytogenetic responses were observed in 66% and 60% of the patients, respectively. Fifty-one of 114 (45%) patients achieved a complete cytogenetic response. No response or progress of disease was noted in 22 out of evaluable 91 patients. The observation period was limited to a maximum of 28 months. A significant improvement in donor chimerism was frequently observed. Only five cases of significant GVHD were reported. Preliminary results show that imatinib mesylate has the potential to positively influence the ratio of donor and recipient cells without inducing a high incidence of severe GVHD. The data suggest that earlier start of imatinib mesylate prior to hematologic relapse in minimum residual disease (MRD) positive patients is a promising treatment concept. PMID- 14529152 TI - Angiographic manifestations and operative findings with 70 cases of hemifacial spasm: relation of common trunk anomalies. AB - The relations between angiographic manifestations and operative findings of hemifacial spasm were studied in 70 cases between 1988 and 2001. Vertebral angiography was performed, and Towne, straight AP, and lateral projections were routinely examined. The dominant anterior inferior cerebellar artery (AICA) directly compressed the facial nerve root exit zone in 26 cases, the dominant posterior inferior cerebellar artery (PICA) in 20, the AICA in 13, the PICA in 2, and the vertebral artery (VA) in 9. Compression by multiple vessels was observed in 11 cases. Anatomical variations of the affected AICA and PICA were classified into 3 groups according to their origins and distributions of blood supply: normal distribution of AICA and PICA in 18%, common trunk anomaly with dominant AICA (basilar artery origin) in 48% and common trunk anomaly with dominant PICA (vertebral artery origin) in 34%. Analyses of the angiograms revealed significantly increased numbers of common trunk anomalies compared with cases with normal angiograms. In 18 of the 20 cases of unilateral common trunk anomalies, facial nerves were compressed by the dominant artery. Preoperative vertebral angiograms may clarify the offending vessels and their sites in most hemifacial spasm cases, thus increasing the safety of surgical interventions. PMID- 14529153 TI - Functional genetic dissection of nuclear receptor signalling in obesity, diabetes and liver regeneration using spatiotemporally controlled somatic mutagenesis in the mouse. AB - The mouse is an excellent animal model for defining human diseases. The null allele mutations (knockouts, KO) have already provided valuable information about their functions, but have also revealed major complications and difficulties: (1) an early embryonic lethality, (2) temporal effect (developmental stage or adult stage), (3) functional redundancy and (4) spatio-effect (cell-autonomous or non autonomous). To overcome these limitations, spatio-temporally controlled somatic mutagenesis, Cre-ER(T)/LoxP system, was established. The nuclear receptors (NRs) play central roles in development, organogenesis, metabolism and energy homeostasis through their ability to transduce hormonal signals into modulation of gene activity. Obesity, excess energy storage in adipose tissue, has a strong link to diabetes. Among NRs, retinoid X receptor alpha (RXRalpha)-peroxisome proliferator-activated receptor gamma (PPARgamma) heterodimers can mediate adipocyte differentiation and obesity which has been demonstrated with in vitro cell culture systems and RXR- and PPARgamma-specific ligand studies. Therefore an adipocyte-specific temporally controlled somatic mutagenesis system was established and analysed. Furthermore, the functional roles of NRs to control liver regeneration were also studied with similar system in hepatocytes. PMID- 14529154 TI - Mitral valve replacement for children with a small annulus using ATS open pivot prosthesis. AB - OBJECTIVES: The present study was conducted to review our experience of mitral valve replacement (MVR) in small children with the ATS open pivot prosthesis. METHODS: Between January 1999 and March 2002, 14 patients (7 males and 7 females) underwent a total of 15 ATS MVRs; 16AP in 9, 18AP in 3, and 20AP in 3 occasions. The mean age and body weight at operation were 1.1 +/- 0.7 (range 0.2 to 2.7) years and 6.8 +/- 2.6 (range 2.8 to 12.5) kg, respectively. RESULTS: There were 2 early and 2 late deaths, each non-valve related. Excluding one patient, the orifice size of the prosthesis was larger than the predicted normal size of the mitral valve for the age. The postoperative Doppler echocardiogram demonstrated normal maximal flow velocity across the prosthesis. There was no morbidity among survivors except for the patient who developed a stuck and immobile valve disc 10.2 months after the operation and required re-replacement. CONCLUSIONS: Our experience shows that MVR with the ATS open pivot prosthesis is a promising surgical option for small children. PMID- 14529155 TI - The outcome and criteria for mitral valve surgery in patients with ischemic mitral regurgitation. AB - OBJECTIVES: Mitral valve surgery for the correction of mitral regurgitation (MR) in patients with ischemic heart disease has been associated with poor prognosis. The criteria for selecting an appropriate surgical procedure are not clearly defined. The objectives of this study were to clarify the criteria for mitral valve manipulation and the outcome in patients with ischemic MR. METHODS: Twenty patients with ischemic MR were proposed for surgery. Ten of them (group A) had grade II MR and underwent isolated coronary artery bypass grafting (CABG). The remaining 10 patients with grade III or more MR underwent both CABG and mitral valve repair (group B). Postoperative left ventricular function and outcome were compared. RESULTS: Preoperative left ventricular end-systolic volume index (LVESVI) in group A was significantly smaller, while preoperative ejection fraction (EF) was almost equal between the groups. EF demonstrated a significant improvement in group A postoperatively, but no changes in group B. Two of group B died following surgery. The other patients were successfully treated with surgery which diminished MR. Congestive heart failure (CHF) developed in 2 patients of group B during follow-up and in none of group A. The 5-year freedom from CHF and the 5-year survival rate in group A were significantly greater than those in group B. CONCLUSIONS: Deterioration of mitral valve function, which necessitated mitral valve repair, was more frequent in the impaired and enlarged hearts. Although mitral repair was beneficial for diminishing MR and New York Heart Association, postoperative course in patients with ischemic MR depended on the preoperative LVESVI. PMID- 14529156 TI - A new sterilization technique with balloon-tube thoracostomy for thoracic empyema. AB - OBJECTIVE: Failure or prolongation of treatment for refractory thoracic empyema by the current chest-tube drainage technique is often due to sterilization difficulties. Insufficient sterilization prolongs hospitalization, and is often associated with life-threatening complications and/or additional invasive surgical procedures. A new chest-tube sterilization technique aimed at making it less invasive and shortening the therapy is proposed. METHODS: Following pretreatment for complications including loculation, bronchopleural fistula, or corticated lung, a double-lumen trocar catheter was introduced at the bottom of the empyemic cavity through the lateral chest wall. Then, a Foley balloon urethra catheter was inserted and attached just inside the anterior chest wall at the top of the cavity for the evacuation of intrathoracic air. After irrigation of the cavity with distilled water once or twice, the cavity was completely filled with a bactericidal solution which was left in place for 30-60 minutes, followed by an antibiotic solution for more than 20 hours. RESULTS: Among the five treated post lobectomy or pneumonectomy cases, sterilization was obtained after only one treatment in four cases and after two courses in the other. Catheterization duration from the initial treatment was 2-13 days. Neither recurrence nor treatment-related major complications were observed. CONCLUSIONS: This balloon tube thoracostomy technique is simple, minimally invasive and cost-effective, due to shortening of the treatment time with minimal manpower and equipment requirements. It is thus a promising therapeutic approach to thoracic empyema and has the potential for application to other intrathoracic disorders. PMID- 14529157 TI - Modified stentless porcine valve enhances accelerated cuspal calcification in the juvenile sheep model. AB - OBJECTIVE: Stent mounting of any bioprosthesis, induces a loss of mobility and reduces the effective valve orifice. By contrast, for stentless procedures, the higher surgical technicality remains a major obstacle for many surgeons. In an attempt to facilitate the insertion of the stentless porcine aortic valve (Toronto SPV), we tried to alter the design by lowering the invasive profile at the depth of the sinuses on both coronary sites. This could theoretically facilitate the implant of the modified stentless valve with an easygoing single layer suture at the challenging subcoronary level and make it more attractive for every surgeon. METHODS: Modifications of the standard model were done by lowering the profile at the depth of the sinuses on both coronary sites, whether by plication or excision of the protruding porcine aortic wall at the nadir of each coronary sinus. Nine juvenile sheep underwent implantation of stentless porcine aortic valves in pulmonary position: 3 standard Toronto SPV, 3 plicated Toronto SPV and 3 excised Toronto SPV. In each series, valves were explanted after 3 months. Valves were analyzed. RESULTS: The cusps of standard Toronto SPV were perfectly functioning and pliable, without visible calcification after three months. The calcium content of the cusps was less than those in the plicated and excised Toronto SPV (2.4 +/- 0.7 microg/mg versus 10.8 +/- 5.9 and 6.7 +/- 3.4 microg/mg). In the plicated and excised valves, calcification of the cusp was more pronounced in the commissural region (3.9 +/- 1.9, 29.0 +/- 16.7, 13.8 +/- 9.5 microg/mg in the standard, plicated and excised Toronto SPV, respectively). On the other hand, the aortic wall from the plicated Toronto SPV had more calcium than that from the other groups (53.6 +/- 6.3, 41.2 +/- 7.1, 45.2 +/- 7.4 microg/mg in the plicated, standard and excised Toronto SPV, respectively). CONCLUSIONS: The modification of stentless porcine valve enhanced accelerated cuspal calcification in the commissural region. It accentuated that the correct implantation technique for stentless procedures is extremely important in order to prevent early degeneration. PMID- 14529158 TI - Thymic atypical carcinoid with Cushing's syndrome manifesting in the terminal stage. AB - A 56-year-old man with recurrent thymic atypical carcinoid presented with Cushing's syndrome only in the terminal stage. Eighteen months after the resection of an anterior mediastinal mass, he presented with hypernatremia and hypokalemia with high serum levels of adrenocorticotropic hormone and cortisol, and died two months later. An autopsy demonstrated multiple metastases of the tumor to the mediastinum, lungs, heart, liver, kidneys, bone, pituitary gland, and ocular muscles. The adrenocorticotropic hormone level of the tumor tissue was 26,000 pg/g. PMID- 14529159 TI - Pathological lesions causing pulmonary hypertension after closure of a ventricular septal defect. AB - A 15-year-old boy with a ventricular septal defect, pulmonary hypertension, Down's syndrome, and extremely thickened media (ETM) of the small pulmonary arteries died of heart failure and pulmonary hypertension 13 years after intracardiac repair. Microscopic examination of lung specimens collected prior to the intracardiac repair and at the time of autopsy revealed that the ETM had remained unchanged and that the arteries connected to the vessels with ETM had become severely thickened. The present case shows that even a small percentage of arteries with ETM can cause pulmonary hypertension, and illustrates one of the mechanisms of how pulmonary hypertension can fail to be resolved after intracardiac repair. PMID- 14529160 TI - Solitary fibrous tumor of the pleura with hemothorax at the thoracic apex. AB - A 31-year-old man was referred to our hospital with a diagnosis of a left lung mass and substantial pleural effusion. 1,300 mL of blood was drained. Chest computed tomography (CT) and magnetic resonance imaging (MRI) revealed a hypervascular, round shaped mass, 9 cm in diameter occupying the left thoracic apex. The mass was broad-based, arising from the area of antero-lateral parietal pleura at the level of the thoracic apex and the first rib. The tumor was successfully resected via median sternotomy and left first intercostal thoracotomy without either claviculectomy or claviculotomy. For the complex surgical access and the tumor's hypervascularity, a careful operative procedure was needed to avoid massive bleeding. The tumor was diagnosed microscopically and immunohistochemically as a solitary fibrous tumor of the pleura and did not meet the criteria for malignancy. PMID- 14529161 TI - Emergent surgery for 3 aged patients who refused elective operation for thoracic aortic aneurysm. AB - This report describes 3 aged patients undergoing emergent surgery who refused elective operation for a thoracic aortic aneurysm because of freedom from symptoms attributable to the aneurysm at the time of presentation. A 77-year-old woman with a thoracoabdominal aneurysm 57 mm in diameter at presentation had recurrent hemoptysis 12 months later. A 78-year-old man with a saccular type distal arch aneurysm 64 mm in diameter at presentation was transported with shock and hemothorax 27 months later. Another 82-year-old man with a saccular type distal arch aneurysm 60 mm in diameter at presentation was admitted with severe chest and back pain 36 months later. All of them underwent tube graft replacements of the aneurysm urgently and were discharged on foot. Aged patients with life-threatening events should not be denied surgical intervention because of excessive operative mortality and morbidity, even if they had previously refused elective surgery. PMID- 14529162 TI - Hemangioma in the anterior mediastinum. AB - Anterior mediastinal hemangiomas are very rare neoplasms in mediastinal tumors. A 58-year-old woman was revealed to have a mass measuring 4 x 3 cm in size in the anterior mediastinum with calcification on computed tomography. It was initially suspected to be a thymoma. We performed tumor extirpation in November 1998. The tumor was close to the thymus and slightly adhered to the superior vena cava, ascending aorta and right phrenic nerve, however, it did not invade any surrounding organs. Histopathologically, it was diagnosed to be a venous type hemangioma composed of vessels covered by smooth muscle and a cavernous type hemangioma composed of dilated vessels covered by one layer of endothelial cells. PMID- 14529163 TI - A pseudoaneurysm after repair of thoraco-abdominal aortic aneurysm. AB - We report a 63-year-old female patient developing a pseudoaneurysm three years after patch-plasty of an aneurysm located at the distal thoracic aorta. Redo operation was performed including total replacement of the descending thoracic aorta with reimplantation of distal intercostal arteries using small-caliber interposition grafts. This case presentation underlines the ineffectiveness of patch repair for the treatment of aortic aneurysms. Furthermore, the surgical technique of reimplantation of intercostal arteries using a separate graft is discussed. PMID- 14529164 TI - A peptic ulcer in a reconstructed gastric tube perforating the thoracic aorta after esophageal replacement. AB - The patient was a 64-year-old man who had received subtotal esophagectomy for esophageal cancer reconstructed by a gastric tube via the thoracic route. He was referred to our hospital in a state of shock, with hematemesis. The bleeding site was not detected with studies of emergent endoscopy and angiography. Massive hemorrhage persisting, an immediate emergent operation was performed. However we were not able to locate the site of bleeding during the operation, and unfortunately we failed to save him. Autopsy showed a peptic ulcer in the gastric pedicle had perforated the thoracic aorta. Ulcers occurring in the gastric pedicle are accompanied by a risk of development of perforation of adjacent important visceras. PMID- 14529166 TI - Familial spontaneous pneumothorax in three generations and its HLA. AB - We experienced a case of familial spontaneous pneumothorax in three generations. Six of 13 family members had episodes of spontaneous pneumothorax. It is well established that there are some diseases associated with human leukocyte antigen (HLA). We performed HLA phenotyping for HLA of A, B and C. In our study, we detected the HLA haplotype A2, B61 in three of 4 who had episodes of spontaneous pneumothorax. The HLA haplotype A2, B70 were also detected in three of 4 who had episodes. This suggests that familial spontaneous pneumothorax might have hereditary factors. PMID- 14529167 TI - Repeat surgery for hemolysis 6 years after replacement of the ascending aorta for acute aortic dissection. AB - A 71-year-old woman underwent replacement of the ascending aorta for Type A aortic dissection. After 6 years, she suddenly developed severe hemolytic anemia, and a second operation for replacement of the ascending aorta was performed. Her hemolysis was thought to occur as follows: the proximal ascending aorta of the graft might have gradually expanded until it compressed the graft. The severe hemolysis was thought to be attributable to disturbance of blood flow by a jet of blood at the site of constriction or the reversed inner felt. Such a case as this is very unusual in that the second operation for hemolytic anemia occurred 6 years after the first surgery. PMID- 14529165 TI - Asymptomatic cor triatriatum incidentally revealed in an elderly patient with mitral stenosis. AB - A case of mitral stenosis accompanied by asymptomatic cor triatriatum, underwent surgical correction, including excision of the diaphragm in the left atrium and mitral valve replacement with a 27 mm St. Jude Medical mechanical valve. A preoperative transthoracic echocardiogram disclosed a linear structure in the left atrium which was suspicious for cor triatriatum, which was confirmed by computed tomography (CT scan) and cardiac catheterization. Cardiac catheterization revealed: 1) mitral valve stenosis of 0.9 cm2 of estimated mitral area, 2) division of the left atrium into two chambers by a diaphragm, that might have multiple ostia; blood flow from the common pulmonary venous chamber to the true left atrium seemed to be unrestricted, and all four pulmonary veins opened into the common pulmonary venous chamber, 3) there were no other apparent cardiac anomalies believed to coexist with cor triatriatum. Preoperative examinations is important for determination of operative strategies. PMID- 14529168 TI - Successful one-stage surgical removal of intravenous uterine leiomyomatosis with right heart extension. AB - We report a 49-year-old woman with intravenous and intracardiac extension of uterine leiomyomatosis. After subtotal hysterectomy and salpingo-oophorectomy, an intravenous and intracardiac tumor was removed under normothermic cardiopulmonary bypass without cardiac arrest. Postoperatively, occlusion of the artery supplying the remaining uterine cervical stump was performed by the catheter coiling technique. More than 2 years after surgery, the patient is well without recurrence. PMID- 14529170 TI - Right lung cancer with right aortic arch. AB - We report here a case of right lung cancer with right aortic arch. A 67-year-old woman with right aortic arch was found to have a 3-cm squamous cell carcinoma arising in the right middle lobe. Right middle lobectomy with lymphadenectomy was performed via right thoracotomy. Although a unique operative view required particular attention in identifying nodes in the mediastinum, we were able to perform the operation without specific techniques. PMID- 14529169 TI - Mobilization of the posterior leaflet of the mitral valve for resection of a left ventricular tumor producing carbohydrate antigen 19-9. AB - We report an extremely rare case of a left ventricular tumor producing carbohydrate antigen 19-9 (CA19-9). A 43-year-old man with an episode of right hemiplegia underwent an emergent operation for a left ventricular tumor just beneath the posterior leaflet of the mitral valve. The tumor was successfully resected by mobilization of the posterior leaflet of the mitral valve. The resected specimen was immunoreactive for CA19-9. PMID- 14529171 TI - Aortic root replacement with a stentless bioprosthesis in a patient with giant cell arteritis. AB - A 66-year-old woman was diagnosed with severe aortic regurgitation. Blood studies constantly showed positive C-reactive protein. Aortic valve replacement using a mechanical valve was carried out. Four months after the operation, echocardiogram showed aortic regurgitation due to paravalvular leakage. Reoperation was performed using a stentless aortic root bioprosthesis. The pathological specimen from the aortic wall was consistent with giant cell arteritis. The patient was treated with prednisone with dramatic improvement of systemic symptoms. PMID- 14529172 TI - Medical students' views on thoracic surgery residency programs in a Japanese medical school. AB - There has been a decline in the number of medical students applying for thoracic surgery training programs. We obtained knowledge of medical students' views on thoracic surgery residency programs. After completion of thoracic surgery clerkship, 17 students were asked to fill out questionnaires on first-year thoracic surgery residency programs. The majority of students considered thoracic surgery to be held in high regard by the general public, and felt that the salary was sufficient. However, only one student chose a thoracic surgery training program. The main reason for not applying for thoracic surgery residency was lifestyle issues. The factors in determining career choice included quality of education and work hours. Medical students are likely to select specialties other than thoracic surgery. Since the main factor influencing medical students' career is the quality of education in a residency program, efforts should be made to improve the quality of education. PMID- 14529173 TI - Surgical treatment of traumatic thoracic aorta rupture: a 7-year experience. PMID- 14529174 TI - Opinion: the species problem, can we achieve a universal concept? AB - One of the so called 'species problems' is that no universal concept exists. There is a tendency among microbiologists to criticize the hitherto devised concept. It is considered by some researchers as being too conservative and not suitable to be compared with those for eukaryotes. However, such problem is not only restricted to prokaryotes, but among other taxonomies comparisons seem to be impossible. As it is argued, the underlying cause to this problem is the reductionistic and monistic use of taxonomy. Analyzing the more than 22 devised concepts it seems possible to achieve a universal species concept. However, this might not be pragmatic. For the time being, a pluralistic sense of the species concept might be accepted, and one will have to recognize that any comparison among different taxonomies will be difficult. PMID- 14529175 TI - Improved fluorescence in situ hybridization of individual microbial cells using polynucleotide probes: the network hypothesis. AB - Fluorescence in situ hybridizations using polynucleotide transcript probes (poly FISH) usually exhibit a ring-shaped halo or corona-like fluorescence signal, whereas hybridizations with oligonucleotide probes (oligo-FISH) result in a uniform and evenly distributed fluorescence throughout the cell. The superiority of poly-FISH in comparison to oligo-FISH regarding the signal intensity and the detection of cells with a low ribosome content suggested a further investigation of the possibilities of polynucleotide probes. Poly-FISH has previously only been described for bacterial cells. In the present study it could also be successfully applied to several yeast species. In addition to that the properties of polynucleotide probes were analyzed by using varying probe lengths and concentrations. This led to the formulation of a hypothesis to explain the characteristic "halo" signal observed with polynucleotide probes. This "network hypothesis" suggests the formation of a network of probes based on the secondary structure of the single stranded RNA probes. Due to the limited permeability of the cell envelope, only part of the probe is linked to its intracellular target site, while the remaining part is located-outside the cell and can form a network by hybridizing with single stranded probes, resulting in the ring-shaped fluorescence signal around the cell. The hypothesis was supported by a number of control experiments including in silico and in vitro analysis of the secondary structure of the probes and hybridizations with probes of defined secondary structures. The network concept provides a new basis for a wider application of poly-FISH. PMID- 14529176 TI - Production of acylated homoserine lactones by different serotypes of Vibrio anguillarum both in culture and during infection of rainbow trout. AB - Onehundred and forty-eight out of onehundred and fifty strains of Vibrio anguillarum isolated from vibriosis in Danish marine aquaculture produced bacterial communication signals, acylated homoserine lactones, eliciting a response in the Agrobacterium tumefaciens (pZLR4) monitoring system. One strain, a serotype O4, induced a strong response in the Chromobacterium violaceum (CV026) monitoring system. Profiles of AHLs determined by TLC separation revealed the presence of at least four AHLs and a compound similar to N-3-oxo-decanoyl homoserine lactone (3-oxo-C10-HSL) was present in all strains. The production rate of the presumed 3-oxo-C10-HSL followed the growth rate of V. anguillarum whereas the production rate of a small AHL (Rf value of 0.74) increased faster than the growth rate of V. anguillarum indicating autoinduction. AHLs were produced by all serotypes (O1 to O10) and by non-typable strains. During infection with V. anguillarum, AHLs could be extracted from liver, kidney and muscle of rainbow trout and AHLs were detected both in vitro and in vivo when cell numbers reached 10(7) per ml or gram. Preliminary investigations of interactions between AHLs and the fish immune system were carried out determining oxidative burst of fish macrophages exposed to 3-oxo-C10-HSL. No activation or suppression of the superoxide anion production in the head kidney macrophages was seen when treated with the AHL compound in concentrations of 1 nM-10 microM. Our data show that AHLs are produced by almost all V. anguillarum strains and that no clear pattern relating AHL production to disease or virulence appear. PMID- 14529177 TI - Cloning, sequence analysis and expression of the F1F0-ATPase beta-subunit from wine lactic acid bacteria. AB - The nucleotide sequences of the genes encoding the F1F0-ATPase beta-subunit from Oenococcus oeni, Leuconostoc mesenteroides subsp. mesenteroides, Pediococcus damnosus, Pediococcus parvulus, Lactobacillus brevis and Lactobacillus hilgardii were determined. Their deduced amino acid sequences showed homology values of 79 98%. Data from the alignment and ATPase tree indicated that O. oeni and L. mesenteroides subsp. mesenteroides formed a group well-separated from P. damnosus and P. parvulus and from the group comprises L. brevis and L. hilgardii. The N terminus of the F1F0-ATPase beta-subunit of O. oeni contains a stretch of additional 38 amino acid residues. The catalytic site of the ATPase beta-subunit of the investigated strains is characterized by the two conserved motifs GGAGVGKT and GERTRE. The amplified atpD coding sequences were inserted into the pCRT7/CT TOPO vector using TA-cloning strategy and transformed in Escherichia coli. SDS PAGE and Western blot analyses confirmed that O. oeni has an ATPase beta-subunit protein which is larger in size than the corresponding molecules from the investigated strains. PMID- 14529179 TI - Tsukamurella spumae sp. nov., a novel actinomycete associated with foaming in activated sludge plants. AB - A polyphasic taxonomic study was undertaken to establish the taxonomic position of six representative strains isolated from activated sewage sludge foam. The organisms were found to have chemical and morphological properties consistent with their assignment to the genus Tsukamurella. DNA:DNA relatedness studies showed that five out of the six isolates formed a distinct genomic species, the remaining strain was most closely associated with this taxon. The five isolates had a unique phenotypic profile that served to distinguish them from representatives of the validly described species of Tsukamurella. The combination of the genotypic and phenotypic data indicated that the five strains should be classified as a new species in the genus Tsukamurella. The name proposed for this taxon is Tsukamurella spumae, the type strain is N1171T (= DSM 44.113T = NCIMB 13947T). It was also shown that some of the reference strains were misclassified as Tsukamurella paurometabola. PMID- 14529178 TI - Planococcus rifietensis sp. nov, isolated from algal mat collected from a sulfurous spring in Campania (Italy). AB - The taxomony of strain M8, isolated from algal mat formed at the origin of a sulfurous spring in Rifieto (Savignano Irpino, Campania, Italy), was investigated in a polyphasic approach. The morphological, physiological and genetic characteristics were compared with of Planococcus and Planomicrobium species. The isolate grew optimally at pH 9.0, 1.8 M NaCl at 37 degrees C. The cells were Gram positive cocci that form pairs, tetrads and aggregates of several cells. The isolate was aerobic/microaerophilic and accumulated glycine-betaine, as a major osmolyte, with minor components glutamate and an unknown compound. M8 was able to hydrolyse X-Glc (5-bromo-4-chloro-3-indoyl beta-d-glucopyranoside). The polar lipid profile consisted of phosphatidylglycerol and diphosphatidylglycerol as major components, and phosphocholine as a minor compound. MK8 was the only quinone found and the fatty acid composition was dominated by branched acids, mainly aiC15:0. The G+C content of DNA was 47.9% and its phylogenetic position was established by 16S rRNA gene sequencing as a member of the genus Planococcus. The DNA/DNA similarity of M8 to the type species Planococcus citreus was less than 55%. For this reason and for physiological and chemotaxonomic features, it is proposed to create a new species Planococcus rifietensis sp. nov. PMID- 14529180 TI - Tepidimonas aquatica sp. nov., a new slightly thermophilic beta-proteobacterium isolated from a hot water tank. AB - A bacterial isolate, with an optimum growth temperature of about 50 degrees C, was recovered from a domestic hot water tank in Coimbra. Phylogenetic analysis using 16S rRNA gene sequence indicated that strain CLN-1T is a member of the beta Proteobacteria and represents a new species of the genus Tepidimonas. The major fatty acids of strain CLN-1T are 16:0, 17:0 cyclo and 16:1 omega7c. Ubiquinone 8 is the major respiratory quinone, the major polar lipids are phosphatidylethanolamine, and phosphatidylglycerol. The new isolate is aerobic and facultatively chemolithoheterotrophic. Thiosulfate and tetrathionate are oxidized to sulfate in the presence of a metabolizable carbon source. Strain CLN 1T grows on amino acids and organic acids, but this organism does not assimilate carbohydrates. Glycerol is the only polyol assimilated. Resinic acids, namely abietic acid, dehydroabietic acid and isopimaric acid are not degraded. On the basis of the phylogenetic analyses, physiological and biochemical characteristics, we propose that strain CLN-1T represents a new species for which we offer the name Tepidimonas aquatica. PMID- 14529181 TI - Methylophaga natronica sp. nov., a new alkaliphilic and moderately halophilic, restricted-facultatively methylotrophic bacterium from soda lake of the Southern Transbaikal region. AB - A new, moderately haloalkaliphilic and restricted-facultatively methylotrophic bacterium (strain Bur2T) with the ribulose monophosphate pathway of carbon assimilation is described. The isolate, which utilizes methanol, methylamine and fructose, is an aerobic, Gram-negative, asporogenous, motile short rod multiplying by binary fission. It is auxotrophic for vitamin B12, and requires NaHCO3 or NaCl for growth in alkaline medium. Cellular fatty acids profile consists primarily of straight-chain saturated C16:0, unsaturated C16:1 and C18:1 acids. The major ubiquinone is Q-8. The dominant phospholipids are phosphatidylethanolamine and phosphatidylglycerol. Diphosphatidylglycerol is also present. Optimal growth conditions are 25-29 degrees C, pH 8.5-9.0 and 2-3% (w/v) NaCl. Cells accumulate ectoine and glutamate as the main osmoprotectants. The G + C content of the DNA is 45.0 mol%. Based on 16S rDNA sequence analysis and DNA DNA relatedness (25-35%) with type strains of marine and soda lake methylobacteria belonging to the genus Methylophaga, the novel isolate was classified as a new species of this genus and named Methylophaga natronica (VKM B 2288T). PMID- 14529183 TI - Molecular characterisation of the species of the genus Zygosaccharomyces. AB - The restriction fragments polymorphisms of the mitochondrial DNA and the PCR fragment that comprised the internal transcribes spacers and the 5.8S rRNA gene, together with the electrophoretic karyotypes of 40 strains from the 10 species of the genus Zygosaccharomyces, including the new species Z. lentus were examined. The RFLP's of the ITS-5.8S region showed a specific restriction pattern for each species, including the new species Z. lentus. The only exception were the species Z. cidri and Z. fermentati that produced identical restriction profiles. The electrophoretic chromosome patterns confirmed the differences between the species of this genus, including the phylogenetic closest species Z. cidri and Z. fermentati. They present few chromosomes ranging from 3 bands (4 or 5 chromosomes) for Z. florentinus to 7 bands (8 to 10 chromosomes) for Z. cidri and Z. fermentati. The strain level resolution power of RFLP's of mtDNA of this genus enabled the characterisation of strains from the same species, even where they are isolated from the same substrate. However, in the cases of Z. bailii and Z. lentus, electrophoretic karyotyping there was considerable variation. PMID- 14529182 TI - Polyphasic characterization of Pseudomonas stutzeri CLN100 which simultaneously degrades chloro- and methylaromatics: a new genomovar within the species. AB - Strain CLN100 was isolated after enrichment on mineral medium with chloronaphthalene as the only carbon and energy source. It was able to use simultaneously and productively chloro- and methyl-derivatives of naphthalene and salicylate through a chromosomally encoded meta pathway. Phenotypic, chemotaxonomic and genotypic characterization classified strain CLN100 as a member of the species Pseudomonas stutzeri. DNA-DNA hybridizations, 16S rDNA, gyrB, rpoD sequences, and molecular fingerprinting indicate that strain CLN100 is a representative of a new genomovar (genomovar 10) within the species. PMID- 14529184 TI - 16S-ARDRA, a tool for identification of lactic acid bacteria isolated from grape must and wine. AB - Lactic acid bacteria (LAB) are found in a great variety of habitats, including grape must and wines. There is a close relationship between the species of LAB which develop during fermentation and the eventual quality of the wine. For these reasons analytical techniques allowing fast and reliable identification of wine LAB are needed. In this work a simple and accurate protocol for identifying species of LAB isolated from grape must and wine is presented. This protocol is based on the amplification, directly from colony, of 16S rDNA and later digestion with one of the following restriction enzymes BfaI, MseI and AluI. A sequential use of the three enzymes is proposed to simplify LAB wine identification, first MseI, then BfaI and finally, if necessary, AluI digestion. The technique was able to discriminate 32 of the 36 LAB reference species tested and allowed the identification of 342 isolates from musts and wines. The isolates belonged to the species: Lactobacillus brevis, L. collinoides, L. coryniformis, L. bilgardii, L. mali, L. paracasei, Leuconostoc mesenteroides, Oenococcus oeni, Pediococcus parvulus and P. pentosaceus. PMID- 14529186 TI - The antimicrobial properties of different strains of Lactobacillus spp. isolated from kefir. AB - The characteristics of 58 strains of Lactobacillus spp. isolated from kefir were studied. These strains were tested for adherence to human enterocyte-like Caco-2 cells, resistance to acidic pH and bile acid, antimicrobial activities against enteropathogenic bacteria and inhibition of Salmonella typhimurium attachment to Caco-2 cells. The best probiotic properties were observed in L. acidophilus CYC 10051 and L. kefiranofaciens CYC 10058. L. kefiranofaciens CYC 10058 produced an exopolysaccharide, which revealed that it was closely related to kefiran, a polysaccharide with antitumoral properties. This is the first in vitro study about the antimicrobial characteristics of the Lactobacillus population of kefir. PMID- 14529185 TI - Combining denaturing gradient gel electrophoresis of 16S rDNA V3 region and 16S 23S rDNA spacer region polymorphism analyses for the identification of staphylococci from Italian fermented sausages. AB - Separation of amplified V3 region from 16S rDNA by denaturing gradient gel electrophoresis (PCR-DGGE) and 16S-23S rDNA intergenic spacer region polymorphism (ISR-PCR) analyses were tested as tool for differentiation of staphylococcal strains commonly isolated from fermented sausages. Variable V3 regions of 25 staphylococcal reference strains and 96 wild strains of species belonging to the genera Staphylococcus, Micrococcus and Kocuria were analyzed. PCR-DGGE profiles obtained were species-specific for S. sciuri, S. haemolyticus, S. hominis, S. auricularis, S. condimenti, S. kloosi, S. vitulus, S. succinus, S. pasteuri, S. capitis and S. (Macrococcus) caseolyticus. Moreover, 7 groups could be distinguished gathering the remaining species as result of the separation of the V3 rDNA amplicons in DGGE. Furthermore, the combination of the results obtained by PCR-DGGE and ISR-PCR analyses allowed a clear differentiation of all the staphylococcal species analysed, with exception of the pairs S. equorum-S. cohnii and S. carnosus-S. schleiferi. The suitability of both molecular techniques and of the combination their results for the identification of staphylococci was validated analysing partial nucleotide sequence of the 16S rDNA of a representative number of wild strains. PMID- 14529187 TI - Identification and differentiation of species and strains of Arthrobacter and Microbacterium barkeri isolated from smear cheeses with Amplified Ribosmal DNA Restriction Analysis (ARDRA) and pulsed field gel electrophoresis (PFGE). AB - ARDRA (Amplified Ribosomal-DNA Restriction Analysis) was used to differentiate among species and genera of Arthrobacter and Microbacteria. Species-specific restriction patterns of PCR-products were obtained with NciI for Arthrobacter citreus (DSM 20133T), A. sulfureus (DSM 20167T), A. globiformis (DSM 20124T) and A. nicotianae strains (DSM 20123T, MGE 10D, CA13, CA14, isolate 95293, 95294, and 95299), A. rhombi CCUG 38813T, and CCUG 38812, and Microbacterium barkeri strains (DSM 30123T, MGE 10D, CA12 and CA15, isolate 95292, and isolate 95207). All yellow pigmented coryneforme bacteria isolated from the smear of surface ripened cheeses were identified as either A. nicotianae or M. barkeri strains. Using pulsed field gel electrophoresis (PFGE) strain specific restriction pattern for all Arthrobacter species and Microbacteria tested were obtained with restriction enzymes AscI and SpeI. PMID- 14529188 TI - Isolation and molecular characterization of thiosulfate-oxidizing bacteria from an Italian rice field soil. AB - In rice paddy soils an active cycling of sulfur compounds takes place. To elucidate the diversity of thiosulfate-oxidizing bacteria these organisms were enriched from bulk soil and rice roots by the most probable number method in liquid medium. From the MPN enrichment cultures 21 bacterial strains were isolated on solid mineral medium, and could be further shown to produce sulfate from thiosulfate. These strains were characterized by 16S rDNA analyses. The isolates were affiliated to seven different phylogenetic groups within the alpha- and beta-subclass of Proteobacteria. Two of these phylotypes were already described as S-oxidizers in this environment (Xanthobacter sp. and Bosea sp. related strains), but five groups represented new S-oxidizers in rice field soil. These isolates were closely related to Mesorhizobium loti, to Hydrogenophaga sp., to Delftia sp., to Pandoraea sp. or showed sequence similarity to a strain of Achromobacter sp. PMID- 14529190 TI - Fingerprinting of yeasts at the strain level by differential sensitivity responses to a panel of selected killer toxins. AB - We used differential sensitivities to a panel of twenty-five cell-free crude killer toxins to fingerprint forty-four Saccharomyces cerevisiae strains of different origin and all taxonomically certified by nDNA-nDNA reassociation. Cluster analysis of numerical data obtained by different growth inhibition areas observed in Petri dishes allowed the complete and reproducible discrimination of all S. cerevisiae strains. PMID- 14529189 TI - A catalogue of molecular, physiological and symbiotic properties of soybean nodulating rhizobial strains from different soybean cropping areas of China. AB - We have analysed 198 fast-growing soybean-nodulating rhizobial strains from four different regions of China for the following characteristics: generation time; number of plasmids; lipopolysaccharide (LPS), nodulation factors (LCOs) and PCR profiles; acidification of growth medium; capacity to grow at acid, neutral, and alkaline pH; growth on LC medium; growth at 28 and 37 degrees C; melanin production capacity; Congo red absorption and symbiotic characteristics. These unbiased analyses of a total subset of strains isolated from specific soybean cropping areas (an approach which could be called "strainomics") can be used to answer various biological questions. We illustrate this by a comparison of the molecular characteristics of five strains with interesting symbiotic properties. From this comparison we conclude, for instance, that differences in the efficiency of nitrogen fixation or competitiveness for nodulation of these strains are not apparently related to differences in Nod factor structure. PMID- 14529191 TI - Arginase activity is a useful marker of nitrogen limitation during alcoholic fermentations. AB - Nitrogen deficiency in musts is one of the causes of sluggish or stuck fermentations. In this work we propose that arginase activity determination can be useful for detecting nitrogen starvation early in vinification. CAR1 and YGP1 genes are not specifically induced under conditions of nitrogen starvation. However, a significant increase in the enzymatic activity of arginase, the product of the CAR1 gene, is detected in vinifications carried out with musts containing limiting amounts of nitrogen. Moreover, on adding ammonia to a nitrogen-deficient vinification, even at late stages, this enzymatic activity is repressed, and growth rate is restored simultaneously. We also investigate the role of ethanol toxicity in nitrogen starvation. The results suggest that ethanol produced during vinification or exogenously added up to 8% (v/v) concentration does not cause nitrogen starvation under the conditions tested because arginase activity is not increased. PMID- 14529192 TI - Differences in safety climate between hospital personnel and naval aviators. AB - We compared results of safety climate survey questions from health care respondents with those from naval aviation, a high-reliability organization. Separate surveys containing a subset of 23 similar questions were conducted among employees from 15 hospitals and from naval aviators from 226 squadrons. For each question a "problematic response" was defined that suggested an absence of a safety climate. Overall, the problematic response rate was 5.6% for naval aviators versus 17.5% for hospital personnel (p < .0001). The problematic response was 20.9% in high-hazard hospital domains such as emergency departments and operating rooms. Problematic response among hospital workers was up to 12 times greater than that among aviators on certain questions. Although further research on safety climate in health care is warranted, hospitals may need to make substantial changes to achieve a safety climate consistent with the status of high-reliability organizations. PMID- 14529193 TI - Associations between errors and contributing factors in aircraft maintenance. AB - In recent years cognitive error models have provided insights into the unsafe acts that lead to many accidents in safety-critical environments. Most models of accident causation are based on the notion that human errors occur in the context of contributing factors. However, there is a lack of published information on possible links between specific errors and contributing factors. A total of 619 safety occurrences involving aircraft maintenance were reported using a self completed questionnaire. Of these occurrences, 96% were related to the actions of maintenance personnel. The types of errors that were involved, and the contributing factors associated with those actions, were determined. Each type of error was associated with a particular set of contributing factors and with specific occurrence outcomes. Among the associations were links between memory lapses and fatigue and between rule violations and time pressure. Potential applications of this research include assisting with the design of accident prevention strategies, the estimation of human error probabilities, and the monitoring of organizational safety performance. PMID- 14529194 TI - Designing teams for first-of-a-kind, complex systems using the initial phases of cognitive work analysis: case study. AB - We present a technique for team design based on cognitive work analysis (CWA). We first develop a rationale for this technique by discussing the limitations of conventional approaches for team design in light of the special characteristics of first-of-a-kind, complex systems. We then introduce the CWA-based technique for team design and provide a case study of how we used this technique to design a team for a first-of-a-kind, complex military system during the early stages of its development. In addition to illustrating the CWA-based technique by example, the case study allows us to evaluate the technique. This case study demonstrates that the CWA-based technique for team design is both feasible and useful, although empirical validation of the technique is still necessary. Applications of this work include the design of teams for first-of-a-kind, complex systems in military, medical, and industrial domains. PMID- 14529195 TI - Speed-of-processing and driving simulator training result in improved driving performance. AB - Useful field of view, a measure of processing speed and spatial attention, can be improved with training. We evaluated the effects of this improvement on older adults' driving performance. Elderly adults participated in a speed-of-processing training program (N = 48), a traditional driver training program performed in a driving simulator (N = 22), or a low-risk reference group (N = 25). Before training, immediately after training or an equivalent time delay, and after an 18 month delay each participant was evaluated in a driving simulator and completed a 14-mile (22.5-km) open-road driving evaluation. Speed-of-processing training, but not simulator training, improved a specific measure of useful field of view (UFOV), transferred to some simulator measures, and resulted in fewer dangerous maneuvers during the driving evaluation. The simulator-trained group improved on two driving performance measures: turning into the correct lane and proper signal use. Similar effects were not observed in the speed-of-processing training or low risk reference groups. The persistence of these effects over an 18-month test interval was also evaluated. Actual or potential applications of this research include driver assessment and/or training programs and cognitive intervention programs for older adults. PMID- 14529196 TI - Effects of age, speech rate, and environmental support in using telephone voice menu systems. AB - Age differences in the use of telephone menu systems were investigated in two experiments. Participants from three age groups (younger: 18-39; middle-aged: 40 59; older: 60+ years) were required to obtain specified information or perform some action using simulated voice menu systems. Measures reflecting task performance, menu navigation, and subjective responses were collected. The focus in Experiment 1 was on examining the effect of speech rate. Data from 196 participants indicated age differences in performance, especially for complex problems. There was no effect of speech rate on any of the performance measures. Experiment 2 examined two types of support devices: a screen phone and a graphical aid. Data from 114 participants indicated that the screen phone provided some benefits in navigational efficiency. In terms of performance, the older participants benefited more from the graphical aid and the younger participants performed better with the screen phone. Actual or potential applications of this research include guidance in the use of support devices that can reduce memory demands and the identification of appropriate speech rates. PMID- 14529197 TI - Cognitive demands of collision avoidance in simulated ship control. AB - The study examines the cognitive demands of collision avoidance under a range of maritime scenarios. Operators used a PC-based radar simulator to navigate set courses over 100 6-min trials varying in collision threat and traffic density. Corrective maneuvers were made through the application of standard navigation rules and by using two decision aids (target acquisition and test maneuver). Results showed widespread effects of collision threat in terms of decision aid use, subjective workload, and secondary task performance. Most notably, demand increased markedly over the course of emergency trials, in which collision threat resulted from rule violation by target vessels. The findings are discussed in terms of the comparison between predictable demands (requiring standard course changes) and those involving uncertainty about the others' intentions (involving more intensive monitoring and forced delays in corrective action). The study has relevance for the design of collision avoidance systems, specifically for the use of ecological displays. PMID- 14529198 TI - Measuring the fit between human judgments and automated alerting algorithms: a study of collision detection. AB - Methodologies for assessing human judgment in complex domains are important for the design of both displays that inform judgment and automated systems that suggest judgments. This paper uses the n-system lens model to evaluate the impact of displays on human judgment and to explicitly assess the similarity between human judgments and a set of potential judgment algorithms for use in automated systems. First, the need for and concepts underlying judgment analysis are outlined. Then the n-system lens model and its parameters are formally described. This model is then used to examine a previously conducted study of aircraft collision detection that had been analyzed using standard analysis of variance methods. Our analysis found the same main effects as did the earlier analysis. However, n-system lens model analysis was able to provide greater insight into the information relied upon for judgments and the impact of displays on judgment. Additionally, the analysis was able to identify attributes of human judgments that were--and were not--similar to judgments produced by automated systems. Potential applications of this research include automated aid design and operator training. PMID- 14529199 TI - New alternative methods of analyzing human behavior in cued target acquisition. AB - Target acquisition tasks in natural environments are often augmented by cuing systems that advise human observers during the decision process. With present technological limitations, cuing systems are imperfect, so the question arises whether cuing aids should be implemented under all conditions. We examined target acquisition performance under different levels of task complexity and cuing system reliability. We introduce here two new methods to help define observer behavior trends in cued target acquisition: a quantitative measure of observer search behavior in a temporal sense and a measure of the extent of observer reliance on the cue. We found that observer reliance on the cue correlated with task difficulty and the perceived reliability of the cue. Cuing was generally helpful in complex tasks, whereas cuing reduced performance in easy tasks. Consequently, cuing systems should be implemented only when the task is difficult enough to warrant the intrusion of a cue into the task. Actual or potential applications of this research include the design and implementation of imperfect automated aids dealing with augmented reality. PMID- 14529201 TI - Gaze-contingent multiresolutional displays: an integrative review. AB - Gaze-contingent multiresolutional displays (GCMRDs) center high-resolution information on the user's gaze position, matching the user's area of interest (AOI). Image resolution and details outside the AOI are reduced, lowering the requirements for processing resources and transmission bandwidth in demanding display and imaging applications. This review provides a general framework within which GCMRD research can be integrated, evaluated, and guided. GCMRDs (or "moving windows") are analyzed in terms of (a) the nature of their images (i.e., "multiresolution," "variable resolution," "space variant," or "level of detail"), and (b) the movement of the AOI (i.e., "gaze contingent," "foveated," or "eye slaved"). We also synthesize the known human factors research on GCMRDs and point out important questions for future research and development. Actual or potential applications of this research include flight, medical, and driving simulators; virtual reality; remote piloting and teleoperation; infrared and indirect vision; image transmission and retrieval; telemedicine; video teleconferencing; and artificial vision systems. PMID- 14529200 TI - Effects of load and speed on lumbar vertebral kinematics during lifting motions. AB - This experimental study investigated the effects of load magnitude and movement speed on lumbar vertebral kinematics during lifting task performance. Ten participants performed sagittally symmetric lifting movements with systematically varied load using either a normal or a faster-than-normal speed. Skin-surface markers were strategically placed over the participants' spinous processes and other landmarks representing major body joints and were recorded during the movements by a motion capture system. The center of rotation (COR) locations and segmental movement profiles for lumbar vertebrae L2 to L5 were derived and analyzed. Results suggested that (a) the COR locations and vertebral angular displacement were not significantly affected by the speed or load variation; (b) a faster speed tended to shorten the time to complete the acceleration for all the lumbar vertebrae considered; and (c) the load increase incurred a tendency for the L5 to complete the primary displacement in a briefer time while enduring greater peak acceleration and velocity. The findings lead to a better understanding of the relation between lifting dynamics and spinal motion. Potential applications of this research include the development of more accurate biomechanical models and software tools for depicting spinal motions and quantifying low-back stress. PMID- 14529202 TI - Bimodal displays improve speech comprehension in environments with multiple speakers. AB - Attending to a single voice when multiple voices are present is a challenging but common occurrence. An experiment was conducted to determine (a) whether presenting a video display of the target speaker aided speech comprehension in an environment with competing voices, and (b) whether the "ventriloquism effect" could be used to enhance comprehension, as found by Driver (1996), using ecologically valid stimuli. Participants listened for target words from videos of an actress reading while simultaneously ignoring the voices of 2 to 4 different actresses. Target-word detection declined as participants had to ignore more distracting voices; however, this decline was reduced when a video display of the target speaker was provided. Neither a signal-detection analysis of performance data nor a gaze-contingent analysis revealed a ventriloquism effect. Providing a video display of a speaker when competing voices are present improves comprehension, but obtaining the ventriloquism effect appears elusive in naturalistic circumstances. Actual or potential applications of this research include those circumstances in which a listener must filter a relevant stream of speech from among multiple, competing voices, such as air traffic control and military environments. PMID- 14529203 TI - Weatherwise: evaluation of a cue-based training approach for the recognition of deteriorating weather conditions during flight. AB - Inappropriate and ineffective weather-related decision making continues to account for a significant proportion of general aviation fatalities in the United States and elsewhere. This study details the evaluation of a computer-based training system that was developed to provide visual pilots with the skills necessary to recognize and respond to the cues associated with deteriorating weather conditions during flight. A total of 66 pilots were assigned to one of two groups, and the evaluation process was undertaken at both a self-report and performance level. At the self-report level, the results suggested that pilots were more likely to use the cues following exposure to the training program. From a performance perspective, there is evidence to suggest that cue-based training can improve the timeliness of weather-related decision making during visual flight rules flight. Actual or potential applications of this research include the development of computer-based training systems for fault diagnosis in complex industrial environments. PMID- 14529204 TI - The IPEM code of practice for electron dosimetry for radiotherapy beams of initial energy from 4 to 25 MeV based on an absorbed dose to water calibration. AB - This report contains the recommendations of the Electron Dosimetry Working Party of the UK Institute of Physics and Engineering in Medicine (IPEM). The recommendations consist of a code of practice for electron dosimetry for radiotherapy beams of initial energy from 4 to 25 MeV. The code is based on the absorbed dose to water calibration service for electron beams provided by the UK standards laboratory, the National Physical Laboratory (NPL). This supplies direct N(D,w) calibration factors, traceable to a calorimetric primary standard, at specified reference depths over a range of electron energies up to approximately 20 MeV. Electron beam quality is specified in terms of R(50,D), the depth in water along the beam central axis at which the dose is 50% of the maximum. The reference depth for any given beam at the NPL for chamber calibration and also for measurements for calibration of clinical beams is 0.6R(50.D) - 0.1 cm in water. Designated chambers are graphite-walled Farmer-type cylindrical chambers and the NACP- and Roos-type parallel-plate chambers. The practical code provides methods to determine the absorbed dose to water under reference conditions and also guidance on methods to transfer this dose to non reference points and to other irradiation conditions. It also gives procedures and data for extending up to higher energies above the range where direct calibration factors are currently available. The practical procedures are supplemented by comprehensive appendices giving discussion of the background to the formalism and the sources and values of any data required. The electron dosimetry code improves consistency with the similar UK approach to megavoltage photon dosimetry, in use since 1990. It provides reduced uncertainties, approaching 1% standard uncertainty in optimal conditions, and a simpler formalism than previous air kerma calibration based recommendations for electron dosimetry. PMID- 14529205 TI - A model to evaluate the biological effect induced by the emitted particles from a beta-delayed particle decay beam. AB - Due to their favourable properties such as high dose localization and high RBE heavy-ion beams have attracted increasing interest in cancer treatment. Efforts to exploit these advantages to the maximum extent in cancer therapy have never been given up. A new idea of applying a radioactive ion beam with beta-delayed particle decay such as 9C or 8B to cancer therapy is put forward in this paper. A model to evaluate the biological effect in terms of cell survival induced by the emitted particles from the decays of the stopped ions has been established. Because of the difference of the internally emitted particle irradiation from the external ion beam, the microdosimetric quantity such as specific energy is applied to evaluate the cell surviving effect induced by the emitted particles from the decays of the radioactive ions. Within the framework of this model, the cell-killing effects resulting from the emitted particles were calculated under different conditions. Finally, the potential application of the radioactive ion beam 9C in cancer therapy is demonstrated. PMID- 14529206 TI - Segment-based dose optimization using a genetic algorithm. AB - Intensity modulated radiation therapy (IMRT) inverse planning is conventionally done in two steps. Firstly, the intensity maps of the treatment beams are optimized using a dose optimization algorithm. Each of them is then decomposed into a number of segments using a leaf-sequencing algorithm for delivery. An alternative approach is to pre-assign a fixed number of field apertures and optimize directly the shapes and weights of the apertures. While the latter approach has the advantage of eliminating the leaf-sequencing step, the optimization of aperture shapes is less straightforward than that of beamlet based optimization because of the complex dependence of the dose on the field shapes, and their weights. In this work we report a genetic algorithm for segment based optimization. Different from a gradient iterative approach or simulated annealing, the algorithm finds the optimum solution from a population of candidate plans. In this technique, each solution is encoded using three chromosomes: one for the position of the left-bank leaves of each segment, the second for the position of the right-bank and the third for the weights of the segments defined by the first two chromosomes. The convergence towards the optimum is realized by crossover and mutation operators that ensure proper exchange of information between the three chromosomes of all the solutions in the population. The algorithm is applied to a phantom and a prostate case and the results are compared with those obtained using beamlet-based optimization. The main conclusion drawn from this study is that the genetic optimization of segment shapes and weights can produce highly conformal dose distribution. In addition, our study also confirms previous findings that fewer segments are generally needed to generate plans that are comparable with the plans obtained using beamlet-based optimization. Thus the technique may have useful applications in facilitating IMRT treatment planning. PMID- 14529207 TI - Non-coplanar beam direction optimization for intensity-modulated radiotherapy. AB - An algorithm for the optimization of the direction of intensity-modulated beams is presented. Although the global optimum dose distribution cannot be predicted, usually a large number of equivalent beam configurations exists. This degeneracy facilitates beam direction optimization (BDO) through a number of possible approximations and because the target set of good beam configurations is very large. Usually, the target volume is accessible through a finite number of paths of little resistance, which are defined by the properties of the objective function and the global optimum dose distribution. Since these paths can be occupied by a finite number of beams, it is reasonable to assume that a minimum number of beams for a configuration that is degenerate to the global optimum exists. Efficiency of the BDO will be characterized by detecting this degeneracy threshold. Beam configurations are altered by adding and deleting beams. A fast exhaustive (up to 3500 non-coplanar orientations) search finds beam directions that improve a configuration. Redundant beams of a configuration can be identified by a fast criterion based on second-order derivative information of the objective function. This offers a fast means of iteratively substituting redundant beams from a configuration. Inferior stationary states can be evaded by adding more beams than the desired number to the current configuration, followed by the subsequent cancellation of superfluous beams. The significance of BDO is examined in a coplanar and a non-coplanar test case. The existence of a threshold number for the minimum configuration and its dependence on the complexity of the problem are shown. BDO outperforms manual configurations and equispaced coplanar beam arrangements in both example cases. PMID- 14529208 TI - Monte Carlo simulations of a scintillation camera using GATE: validation and application modelling. AB - Geant4 application for tomographic emission (GATE) is a recently developed simulation platform based on Geant4, specifically designed for PET and SPECT studies. In this paper we present validation results of GATE based on the comparison of simulations against experimental data, acquired with a standard SPECT camera. The most important components of the scintillation camera were modelled. The photoelectric effect. Compton and Rayleigh scatter are included in the gamma transport process. Special attention was paid to the processes involved in the collimator: scatter, penetration and lead fluorescence. A LEHR and a MEGP collimator were modelled as closely as possible to their shape and dimensions. In the validation study, we compared the simulated and measured energy spectra of different isotopes: 99mTc, 22Na, 57Co and 67Ga. The sensitivity was evaluated by using sources at varying distances from the detector surface. Scatter component analysis was performed in different energy windows at different distances from the detector and for different attenuation geometries. Spatial resolution was evaluated using a 99mTc source at various distances. Overall results showed very good agreement between the acquisitions and the simulations. The clinical usefulness of GATE depends on its ability to use voxelized datasets. Therefore, a clinical extension was written so that digital patient data can be read in by the simulator as a source distribution or as an attenuating geometry. Following this validation we modelled two additional camera designs: the Beacon transmission device for attenuation correction and the Solstice scanner prototype with a rotating collimator. For the first setup a scatter analysis was performed and for the latter design. the simulated sensitivity results were compared against theoretical predictions. Both case studies demonstrated the flexibility and accuracy of GATE and exemplified its potential benefits in protocol optimization and in system design. PMID- 14529209 TI - The effect of water molecular self-diffusion on quantitative high-resolution MRI polymer gel dosimetry. AB - In polymer gel dosimetry, magnetic resonance imaging (MRI) is used to determine the spin-spin relaxation rate (R2) which in turn can be correlated with absorbed dose to provide a map of the spatial distribution of the absorbed dose in the irradiated dosimeter. High accuracy, precision and reproducibility of these dose maps are essential. Moreover, for dose verification around brachytherapy sources used for intravascular brachytherapy, a high spatial resolution is required (typically 0.01-0.1 mm). To achieve these microscopic resolutions, strong imaging gradients are applied. The Brownian motion of water molecules in the presence of these strong magnetic field gradients causes an attenuation of the MR signal. When using a multiple spin-echo sequence, this may result in a significant deviation in the measured R2. The diffusion-related change in R2 at high resolutions was investigated experimentally and correlated with predictions that were obtained numerically and algebraically. Diffusion weighting is determined by the self-diffusion coefficient D, and imaging parameters, quantified by the b factor. The b-factor was calculated for a multiple spin-echo sequence for different gradient strengths and gradient pulse durations. The variations in R2 that were observed when changing the matrix size and slice thickness are explained. It is shown that a linear correlation between the matrix size and the variation in R2 is based on the diffusion weighting caused by the read-out gradients and slice selective gradients. In conclusion, the essence of taking into account molecular self-diffusion to quantify variations in the measured dose R2 response when using high-resolution MRI in polymer gel dosimetry is emphasized. PMID- 14529210 TI - Medical personnel and patient dosimetry during coronary angiography and intervention. AB - Percutaneous coronary interventions are associated with increased radiation exposure compared to most radiological examinations. This prospective study aimed at (1) measuring entrance doses for all in-room personnel, (2) performing an assessment of patient effective dose and intracoronary doses, (3) investigating the contribution of each projection to kerma-area product (KAP) and irradiation time, (4) comparing results with established DRL values in this clinical setting and (5) estimating the risk for fatal cancer to patients and operators. Measurements were performed during 40 consecutive procedures of coronary angiography (CA), half of which were followed by ad hoc coronary angioplasty (PTCA). KAP measurements were used for patients and thermoluminescent dosimetry for the in-room personnel. The mean KAP value per procedure for CA was 29 +/- 9 Gy cm2. Thirty four per cent of KAP was due to fluoroscopy, whereas the remainder (66%) was due to digital cine. Accordingly, the mean KAP value per PTCA procedure was 75 +/- 30 Gy cm2, and contribution of fluoroscopy is 57%. Effective dose per year was estimated to be 0.04-0.05 mSv y(-1) for the primary operator, and 0.03 0.04 mSv y(-1) for those assisting. Corresponding measurements for radiographer and nurse were below detectable level, implying minimal radiation hazards for them. Regarding radiation exposure, coronary intervention is considered a quite safe procedure for both patients and personnel in laboratories with modern equipment and experienced operators as long as standard safety precautions are considered. Exposure optimization though should be constantly sought through continuous review of procedures. PMID- 14529211 TI - Helical cardiac cone beam reconstruction using retrospective ECG gating. AB - In modern computer tomography (CT) systems, the fast rotating gantry and the increased detector width enable 3D imaging of the heart. Cardiac volume CT has a high potential for non-invasive coronary angiography with high spatial resolution and short scan time. Due to the increased detector width, true cone beam reconstruction methods are needed instead of adapted 2D reconstruction schemes. In this paper, the extended cardiac reconstruction method is introduced. It integrates the idea of retrospectively gated cardiac reconstruction for helical data acquisition into a cone beam reconstruction framework. It leads to an efficient and flexible algorithmic scheme for the reconstruction of single- and multi-phase cardiac volume datasets. The method automatically adapts the number of cardiac cycles used for the reconstruction. The cone beam geometry is fully taken into account during the reconstruction process. Within this paper, results are presented on patient datasets which have been acquired using a 16-slice cone beam CT system. PMID- 14529212 TI - Functional CT imaging of prostate cancer. AB - The purpose of this paper is to investigate the distribution of blood flow (F), mean capillary transit time (Tc), capillary permeability (PS) and blood volume (vb) in prostate cancer using contrast-enhanced CT. Nine stage T2-T3 prostate cancer patients were enrolled in the study. Following bolus injection of a contrast agent, a time series of CT images of the prostate was acquired. Functional maps showing the distribution of F, Tc, PS and vb within the prostate were generated using a distributed parameter tracer kinetic model, the adiabatic approximation to the tissue homogeneity model. The precision of the maps was assessed using covariance matrix analysis. Finally, maps were compared to the findings of standard clinical investigations. Eight of the functional maps demonstrated regions of increased F, PS and vb, the locations of which were consistent with the results of standard clinical investigations. However, model parameters other than F could only be measured precisely within regions of high F. In conclusion functional CT images of cancer-containing prostate glands demonstrate regions of elevated F, PS and Vb. However, caution should be used when applying a complex tracer kinetic model to the study of prostate cancer since not all parameters can be measured precisely in all areas. PMID- 14529214 TI - On the inclusion of proliferation in tumour control probability calculations for inhomogeneously irradiated tumours. AB - In previous modelling of tumour control probability (TCP) for inhomogeneously irradiated tumours we used an expression that did not include a proliferation correction term, which should be lambda(T - Tk). We did not use that term in the specific examples in our previous work to model slowly growing tumours in order to avoid unnecessary mathematical complexity. We have now considered how to do so in more detail, and there are some variations, such as schedules that depart from a number of equal fractions over the entire course of treatment, if one wishes to compensate for proliferation in the remaining fractions by increasing the dose per fraction after the kick-off time has passed in order to achieve the same TCP when proliferation is neglected. PMID- 14529213 TI - Monte Carlo study of TLD measurements in air cavities. AB - Thermoluminescent dosimeters (TLDs) are used for verification of the delivered dose during IMRT treatment of head and neck carcinomas. The TLDs are put into a plastic tube, which is placed in the nasal cavities through the treated volume. In this study, the dose distribution to a phantom having a cylindrical air cavity containing a tube was calculated by Monte Carlo methods and the results were compared with data from a treatment planning system (TPS) to evaluate the accuracy of the TLD measurements. The phantom was defined in the DOSXYZnrc Monte Carlo code and calculations were performed with 6 MV fields, with the TLD tube placed at different positions within the cylindrical air cavity. A similar phantom was defined in the pencil beam based TPS. Differences between the Monte Carlo and the TPS calculations of the absorbed dose to the TLD tube were found to be small for an open symmetrical field. For a half-beam field through the air cavity, there was a larger discrepancy. Furthermore, dose profiles through the cylindrical air cavity show, as expected, that the treatment planning system overestimates the absorbed dose in the air cavity. This study shows that when using an open symmetrical field, Monte Carlo calculations of absorbed doses to a TLD tube in a cylindrical air cavity give results comparable to a pencil beam based treatment planning system. PMID- 14529215 TI - Arrow injuries in North East Nigeria. AB - Arrow injuries are an extinct form of injury in most parts of the developed world but constitute 0.1% of emergency admissions in the University of Maiduguri Teaching Hospital annually. This is a retrospective study of arrow injuries carried out over a ten-year period (1989-1999) in order to study the reasons for its continued incidence the presentation and the experiences in the management of these injuries. There were 73 cases of arrow injuries and were all males with a peak age incidence of 31-40 years of age. Majority were farmers, cattle herdsmen and traders from the northeast region of Nigeria and the neighbouring republic of Cameroon, Chad and Niger. The various reasons for the injuries were armed robbery in 41%, communal clashes 20.5%, dispute between farmers and cattle herdsmen 13%, and cattle theft 8%. Majority were clinically stable on presentation with arrows in the head, neck, chest and abdomen this resulted in various surgical procedures in order to remove the arrows and repair damaged viscera. Unstable presentations resulted in mortalities (4.1%) preoperatively. Wound infection was the most common complication in patients who presented late. Improvements in the socio economic conditions in the region and legislation on the use of these weapons would reduce the incidence of these injuries. PMID- 14529216 TI - Antimicrobial sensitivity pattern of organisms causing urinary tract infection in children with sickle cell anaemia in Ibadan, Nigeria. AB - As part of a larger project on childhood urinary tract infection, antimicrobial sensitivity tests were carried out on the bacterial isolates from the urine of febrile children seen at the University College Hospital, Ibadan, Nigeria. METHODOLOGY: Midstream urine specimens were collected from 171 sickle cell anaemia children and from an equal number of haemoglobin-A controls and cultured by standard methods. Sensitivity to eleven antimicrobials was tested using the disc-diffusion technique of Stokes. RESULTS: Significant bacteriuria was obtained from 37 children with sickle cell anemia and 27 controls. The isolates were Escherichia coli, Klebsiella species, Non-haemolytic streptococcus, beta haemolytic Streptococcus, Salmonella, Proteus and Pseudomonas species. Sensitivity was highest to Pefloxacin to which over 94% of the organisms were sensitive followed by Ceftriaxone (over 85%) and ceftazidime (over 85%). Sensitivities to nalidixic acid and cefuroxime were between 67.6% and 74.1%. Most of the isolates were resistant to gentamicin, amoxycillin, cotrimoxazole and ampicillin. In general the sensitivity pattern in the sickle cell anaemia group was similar to the pattern in the control group. CONCLUSION: Aetiological agents of childhood UTI in this environment are resistant to most of the drugs commonly recommended for its treatment. Nalixidic acid and cefuroxime are recommended as first line drugs while awaiting results of sensitivity testing. Ceftriazone and ceftazidime should be reserved for cases of non-response to first line drugs and in severe cases. Pefloxacin should be considered potential drug of treatment particularly in multi-drug resistant infections. PMID- 14529217 TI - Management outcome of pulmonary tuberculosis: a nine year review in Ilorin. AB - OBJECTIVE: This study evaluates the management outcome of patients with pulmonary tuberculosis (PTB) over a nine year period. Essentially the cure rate, rate of default and fatality rate were all estimated. Factors predisposing to poor drug compliance were also reviewed. METHODS: This was conducted by analysing the medical records of all newly diagnosed cases of PTB. Patients' demographic data, their social and past medical histories were obtained from their records. The extents of the lung lesions were determined from their chest X-rays. Criteria were set to define those that were (a) cured, (b) defaulted (c) transferred (d) had treatment failure and (e) died. The chi-square test of significance was used to estimate the difference between those that were cured and those that defaulted therapy. Multiple regression models were used to determine the level of interplay among the various factors contributing to patients' poor therapy compliance; the best of these was recorded. RESULTS: Out of the 1,741 cases of PTB treated over the studied period, 43.7% were cured, 0.3% had treatment failure, 44.2% defaulted therapy, 0.2% were transferred to other treatment centers while a fatality rate of 11.6% was recorded. Male gender, old age (> or =) 65 years), unmarried status, and previous default from therapy were found associated with poor therapy compliance. Others were unemployment, cigarette smoking and the use of alcohol. CONCLUSION: The cure rate of PTB in this study was very low, while the rate of default from therapy was quite high. These are in contrast to WHO target for TB control. To improve on these, directly observed therapy short course (DOTS) strategy for treating TB has to be adopted and implemented at all our treatment points for PTB. There should be emphasis on the Primary health Care (PHC) centers, because majority of PTB patients are grass root people and PHC set-ups are closer to them. PMID- 14529218 TI - Testicular torsion on the Jos Plateau. AB - In order to determine the pattern and the factors that influenced outcome, we retrospectively studied fifty-seven patients with torsion of the testis admitted to the Jos University Teaching hospital between August 1993 and July 2001. The age ranged from 2 to 55 years with a mean of 22.7 years. Majority (79%) of the patients were in the second and third decades of life. The main suspected precipitating factors in this study were cold weather and scrotal trauma: in 28% of the cases no cause could be ascertained. Eight (14%) patients presented within 4 hours and 35 (61%) presented after 24 hours of the onset of symptoms. Both sides were equally affected. Testicular pain, retraction and scrotal swelling were the most common presenting complaints. The highest incidence 65%) occurred between November and February when the weather on the Jos plateau is coldest. At surgery, 34 (60%) patients were found to have associated congenital anomalies; in 22 (39%) patients, the testis was non-viable. There was no mortality in this series and the complications were superficial wound infection (14%), testicular atrophy (7%) and sub-fertility (16%). High index of suspicion in a patient with acute scrotum, prompt and effective surgery will improve testicular salvage. PMID- 14529219 TI - Uses and misuse of blood transfusion in obstetrics in Lagos, Nigeria. AB - The objectives of this study were to identify obstetric and medical conditions in transfused patients, appraise the justification for the transfusion and recommend measures for reducing transfusion in obstetrics in Nigeria. Available case records of obstetric patients transfused at the Lagos University Teaching Hospital between the period 1st November 1995 and 31 October 1998 were reviewed retrospectively. Data collected included booking status, mode of delivery, amount of blood-transfused, post transfusion packed cell volume (PCV), and obstetric and medical conditions found in the patients. There were 4,159 cases out of which 503 were transfused. The overall transfusion rate was 12.1%, in booked and unbooked patients, it was 6.6% and 45.8% respectively. Only 231 cases were available for full analysis, mean age was 28.9 +/- 5.2 years (range 16-43 years). Sixty-three (27.3%) had unit-transfusions. Mean post transfusion PCV was 28.4% +/- 4.3% (range 17%-43%). In 63 (27.3%), the post transfusion PCV was above 30%. Some of the obstetric and medical conditions in the transfused patients were caesarean section (68.8%), previous caesarean section (20.4%), antepartum haemorrhage (16.9%), pregnancy induced hypertension (15.6%), anaemia and malaria (14.0%), induction of labour (13.0%), ruptured uterus (8.8%), and sickle cell anaemia (5.2%). Multi-unit transfusions were found in ruptured uterus (5.8 units), sickle cell anaemia (4.9 units), vaginal and cervical lacerations (4.0 units), forceps delivery (3.9 units) and malaria and anaemia (2.9 units). CONCLUSION: The study shows a high transfusion rate; an appreciable number were unnecessary transfusions. A number of the obstetric and medical factors for blood transfusion were avoidable. A reduction in blood transfusion rate can be achieved by the provision of adequate, available and affordable maternal health services in Nigeria. PMID- 14529220 TI - Elective non-instrumented anterior cervical diskectomy and fusion in Ghana: a preliminary report. AB - BACKGROUND: This study is a retrospective analysis of forty-one consecutive patients who underwent elective single or multilevel anterior cervical diskectomy and fusion (ACDF) in Ghana. All the patients had been followed up for at least six months. METHODS: The medical records of forty-one consecutive cases were analysed retrospectively. The parameters reviewed included patient demographics and presentations, number of fusion levels, complications and clinical outcome. RESULT: Non-instrumented ACDF constituted 22% of all the neurosurgical procedures performed during the study period. A total of 41 patients underwent the procedure. Seventy-eight percent of the patients were male; the mean age of all the patients was 52 years; follow-up was for an average of 21 months. Preoperative assessment revealed that 98% had myelopathy and 2% had radiculopathy only. No patient was operated on for only pain. There were no repeat operations performed. All levels operated on were fused for a total of 67 levels; 37% at one level only and 63% at two levels; no patient was fused or operated on at three levels. Ninety percent of the fusions were at the C4-5 and C5-6 levels. The mean ages of males and females fused at one level only were 48 and 60 years respectively; the difference was statistically significant (P<0.05). For patients fused at two levels; 81% were males and 19% were females, the difference was statistically significant (P<0.05). However, there was no statistical difference between the ages of males and females operated on at two levels (P>0.05). In addition there was no statistical difference between the ages of males operated on at one level or two levels nor was there a difference between the ages of females operated on at one or two levels (P>0.05). The mean preoperative Nurick grade was 2.3 (SD, 1.9); the mean Nurick grade postoperation was 1.3 (SD, 1.3). The difference is significant (P<0.001). There was also a significant relationship between the preoperative and postoperative Nurick grades for males operated on at either one or two levels (P<0.01); however none could be demonstrated for females (P>0.01). The total operative complication rate was 12%; the most common complication was graft/donor site infection (7%). There was no operative or postoperative mortality. Eighty six percent of the patients had an excellent or good clinical outcome as defined by Odom's criteria. Of the 34% who did not have excellent or good clinical outcome, follow up radiologic studies showed excellent graft fusion in all of them. CONCLUSION: The majority of patients undergoing non-instrumented ACDF in Ghana have cervical myelopathy involving two cervical levels. However, a large majority of them have excellent or good clinical results after surgery. The most common complication is graft/donor site infection. PMID- 14529221 TI - Delayed primary realignment of posterior urethral rupture. AB - The treatment of acute posterior urethral rupture is controversial. Twelve patients who presented with acute posterior urethral rupture over a five-year period were treated by delayed primary realignment of the injury. The technique of this procedure and the outcome are the subject of this presentation. Eight patients had successful realignment without strictures. Two patients with strictures responded to simple dilatations with bougies and the other two had formal urethroplasty. PMID- 14529222 TI - Herpes zoster infection and HIV seropositivity among eye patients--University of Ilorin Teaching Hospital experience. AB - This paper reports cases of Herpes Zoster Ophthalmicus (HZO) seen in 10 Nigerian adults at the Eye clinic of the University of Ilorin Teaching Hospital (UITH), Ilorin, Kwara State, Nigeria, some of whom tested positive to HIV infection using ELISA method with confirmation using the Western blot test. There were 6 female and 4 male patients. Five (50%) of the patients tested positive for HIV. A high index of suspicion should be maintained among Ophthalmologists when confronted with patients with HZO who are healthy looking. PMID- 14529223 TI - Emotional distress and strain in relatives of patients with severe mental disorders: a comparative study. AB - The levels of emotional distress and strain were examined in two groups of relatives of psychiatric patients using standard instruments. The dementia group had more GHQ--30 cases than the schizophrenic group. Even though the schizophrenic relatives had higher distress and strain scores than the dementia relatives, they did not reach significant levels when the two groups were compared. PMID- 14529224 TI - Haemoglobin F levels in healthy Nigerian adults. AB - Sickle Cell disease is a major genetic disorder in tropical Africa; its severity is often ameliorated by the presence of high levels of HbF, which is genetically determined. HbF was assessed in healthy Nigeria adults using the two minutes alkali denaturation method of Betke. The subjects studied included twenty-six males and twenty-four females all with HbA this was compared with twenty heterozygotes (HbAS). The mean HbF of the entire subject studied was 2.5 +/- 1.7% (range 0.4-12.8%). The mean value for Individuals with HbA genotype was 2.7 +/- 3.4% (range 0.4-12.8%). While the AS subject had a mean of 2.4 +/- 2.2% (range 0.7-8.4%). Twenty-two percent of the population studied had value greater than 3%. The high Level of HbF among healthy adults is believed to be genetic and related to the high prevalence of sickle cell disease in this sub-region. The association between HbF and high Persistence of fetal haemoglobin (HPFH) and the thalassaemias are discussed. PMID- 14529225 TI - Influence of positions on the incidence and severity of maxillofacial injuries in vehicular crashes [corrected]. AB - The objective of this study was to determine the influence of positions on the incidence and severity of maxillofacial injuries in vehicular crashes. Two hundred and fifty cases of RTA were seen and studied from October 1999 to May, 2000. They comprised 72.8% males and 27.2% females with a male to female ratio of 2.7:1. The age range was 1 to 80 years with a mean age of 40 years. The most common vehicle involved in RTA was the mini bus, 34.3% followed by motorcycle, 26.5%. Middle seat passengers, drivers and motorcycle passengers and riders sustained more and severe injuries during crashes. The mandible was more frequently fractured at the symphysis (24.6%) than at any other site while the maxilla was frequently fractured at the zygomatic bone and arch (10.8%). More injuries occurred in the age range of 20-30 years and these accounted for 31.8% of the cases. Only 6 occupants of motor vehicles wore seat belts before accident while no motorcycle rider or passenger wore any helmet. In conclusion, drivers, middle seat occupants of a vehicle, motorcycle passenger and riders sustained more and severe injuries than back seat occupants. PMID- 14529226 TI - A 3-year review of neurologic admissions in University College Hospital Ibadan, Nigeria. AB - A 3 year review of neurologic admissions into the adult medical wards at the UCH, Ibadan, Nigeria between January 1998 and December 2000 is presented. The study design involved the scrutiny of the records of all the neurological admissions, male and female to the medical ward. The identified cases were then classified and only cases confirmed as neurological were further analysed. Stroke, predominantly non-hemorrhagic accounted for 50.4% of cases for the period of study. Stroke is therefore the most common cause of adult neurologic admissions on medical wards of UCH. Central nervous system infections, comprising mainly of tetanus and meningitis accounted for 14.2% (111) and 12.4% (97) of case respectively. The myelopathies were the cause of neurologic admissions in 8.1% (63) of cases followed by seizure disorders. Headache was the reason for admission in 0.9% (7) of cases. Parkinsons disease, hypertensive encephalopathy, Guillian Barne syndrome, seasonal ataxic neuropathy, cavernous sinus thrombophlebitis, normal pressure hydrocephalus were rarely the cause of admission. Similarly, dystonia, and cerebral malaria recorded 0.13% (1) of cases each. A case is made for the establishment of regional stroke units in Nigeria. PMID- 14529227 TI - Clinical diabetic nephropathy in a tropical African population. AB - Diabetic nephropathy is the single most important disorder leading to renal failure in adults in the Western countries and it is among the first three major causes of end stage renal disease in Nigeria. The aim of this study is to show the features of clinical diabetic nephropathy in the Olabisi Onabanjo University Teaching Hospital, Ogun State, Nigeria. The study group consists of 342 consecutive diabetic patients with persistent proteinuria (positive albustix) and diabetic retinopathy, seen from January 2000 to June 2001 in the Ogun State University Teaching Hospital, Sagamu. Clinical and laboratory parameters were recorded. Students' t-test and Spearman correlation coefficient were used in analysis. The frequency of occurrence of clinical diabetic nephropathy is 28.4% with majority already symptomatic despite normal biochemistry. Mean ages of type 1 and type 2 are 26+/-7.9 years and 53.4+/-6.3 years respectively with a sex ratio of 1.2:1. Mean duration of disease is 6.5+/-3.6 years and 9.4+/-4.1 years respectively. Two hundred and seventy-one (79.2%) patients were hypertensive. Nephrotic syndrome is not a common presentation. Diabetic nephropathy is a significant problem in this environment. PMID- 14529228 TI - The place of clinical features and standard chest radiography in evaluation of mediastinal masses. AB - One hundred and five primary mediastinal masses were seen between 1975 and 1998, at the Cardiothoracic surgical Unit of the University College Hospital Ibadan. These were studied to establish the importance of clinical features and plain chest radiography in preoperative evaluation of these masses. The sources of information were unit's record, cancer registry and the patients case note. The age range of the 75 male patients was 3 to 80 years (mean 35.2 +/- 22.1 year) and for the 30 female patients was 2.5 to 70 years (mean 30.9 +/- 18.6 years). Anterosuperior mediastinal masses were the most common (63.8%) followed by posterior mediastinal masses (22.9%). Middle mediastinal masses made up the remaining (13.3%). The most common primary mediastinal tumour was lymphoma (21.9%) next to which were thymus gland neoplasia and thymus cysts/hyperplasia (18.1%). Endocrine tumours (mainly goiters) constituted 17.1% of the masses. On the whole, 45 (42.9%) of the mediastinal tumours were malignant and 60 (57.1%) were benign. Eighty one patients (77.1%) were symptomatic at presentation, 24 patients (22.9%) were asymptomatic. Malignancy was more associated with symptoms in this series (82.2%) and benign lesion were more frequently (70.8%) asymptomatic. Cervical lymphadenopathy (78.6%), was more frequent in patients with malignant lesion. Tracheal deviation (60.3%) and neurological signs (78.9%) were more frequent in benign disease. Superior vena cava syndrome was more frequently associated with primary malignant mediastinal tumours. All masses (100%) were visualized on plain chest x-ray. On the basis of clinical features and chest x-ray, majority of patients (76.2%) with primary mediastinal masses had exploratory thoracotomy, sternotomy or biopsy of their mediastinal mass. PMID- 14529229 TI - A questionnaire survey of senior house officers/registrars response to their training at University College Hospital, Ibadan. AB - This study evaluated the responses of randomly selected 54 senior house officers (SHO)/registrars to questions on their training at the University College Hospital, Ibadan, Nigeria. Previous studies have identified excess workload, poor supervision and inadequate hours of formal educations as areas deserving increased attention. The aim of this study is to assess the residents perception of their training and to offer recommendations where necessary. Many residents felt they are often called to attend to problems beyond their experiences and competence. As many as 66.7% as at 3 months and above into training, have not had the opportunity to discuss their training progress with their supervising consultants. The study therefore calls for more hours of formal teaching and consultants' regular evaluation of residents training. A regular conduct of auditing of training programmes is recommended. PMID- 14529230 TI - Intraventricular haemorrhage and periventricular leukomalacia in Nigerian infants of very low birth weight. AB - Transfontanelle ultrasound scans were performed on 93 very low birth weight neonates admitted at the University of Ilorin Teaching Hospital (UITH), Ilorin, Nigeria, between January 1992 and April 1994, to ascertain the pattern and determinants of intraventricular haemorrhage and periventricular leukomalacia. Forty-seven percent of the infants had normal ultrasound; 22% had mild intraventricular haemorrhage (IVH), 7.5% had moderate to severe IVH and 23% had periventricular leukomalacia (PVL) and a 3.5 fold increase in IVH. Infants with apgar scores < or = 3 at 5 minutes had a 6-fold increase in IVH and 2-fold increase in PVL. There was no association with birth weight, gestational age, respiratory distress and place of delivery. Our study suggests that antepartum events proximate to delivery and the immediate post partum events play significant roles in the development of IVH and PVL in this population. Improvement in antenatal, labour and delivery, and immediate newborn care; including provision of basic resuscitation equipment and skills to midwives in the rural maternity centers will reduce the incidence and severity of IVH and PVL. PMID- 14529231 TI - A preliminary survey of central nervous system tumors in Tema, Ghana. AB - BACKGROUND: In January 2000, the first ever neurosurgical program in Tema was established. This preliminary survey was conducted for the following purposes. 1) to determine the relative frequencies of the various histopathological types of CNS tumors. 2) To relate the occurrence of the various types of CNS tumors to age, sex, symptoms, neurologic findings and location. 3) to review the current use of neurodiagnostic modalities. METHODS: A retrospective analysis of the records of 30 consecutive patients seen at T. I. N. with histologically proven CNS tumors was carried out. The following parameters were analysed; sex, age, symptoms, neurologic status, surgical procedure, histopathological diagnosis, pre and post operative Karnofsky rating. RESULTS: 30 patients (14M, 16F) constituted the series. Their mean age was 39.8 (R 2-72, SD, 18.7) years. The difference between the mean ages of patients with intracranial or spinal tumors was not significant (P>0.05). For intracranial tumors, there was a significant difference between the mean ages of those with infratentorial and supratentorial tumors. Spinal tumors constituted 13% of the series and they all presented with paraplegia. Eighty seven percent had intracranial tumors; of these 27% presented with headaches and 31% with seizures. Only 62% of patients with intracranial tumors presented with neurologic deficits. CT scanning was the diagnostic modality utilized in the diagnosis of all the intracranial tumors. Cerebral angiography was not obtained in any case. Myelography and post myelography CT scanning diagnosed all spinal tumors. Surgical procedures for CNS tumors constituted 23% of all neurosurgical surgical procedures performed during the study period. All patients with spinal tumors underwent laminectomy only. Sixty five percent of those with intracranial tumors underwent craniotomy; 34% underwent stereotactic biopsy. The most common intracranial tumor was high-grade astrocytoma (HGA), 23%. The left frontal lobe was the most common location of the intracranial tumors (54%), followed by the left temporal lobe (36%). No significant relationship was demonstrated in the brain tumor sites (P>0.05). At presentation, 46% of those with intracranial tumors had a karnofsky rating >70. One week after surgical intervention, this had increased to 62%. There was no significant relationship between tumor histopathology and preoperative or postoperative Karnofsky rating. The mortality rate in the first 10 days following intracranial tumor surgery was 8%; there were no deaths in the spine surgery group. The post operative complication rate for CNS tumor surgery was 11.5%. CONCLUSION: Intracranial tumors are the most common type of CNS tumors in Tema. More than one third of patients with intracranial tumors presented without neurologic deficits. HGA is the most frequently seen intracranial tumor. All patients with spinal tumors presented with neurologic deficits, specifically paraplegia. PMID- 14529232 TI - Orbital metastases of prostatic carcinoma in a tropical African population. AB - We have reviewed the cases of orbital metastases from carcinoma of the prostate gland seen in the University College Hospital, Ibadan over an 11 year period, 1990-2000. During the period, seven cases of orbital metastases that presumably arose from carcinoma of the prostate were seen. Four of the patients died of progression of the primary disease over a variable period ranging between 2 weeks and 30 months of diagnosis and treatment, two were lost to follow up and one is alive 46 months after initial diagnosis of orbital metastases from carcinoma of the prostate and treatment. PMID- 14529233 TI - Epidemiology of trauma deaths. AB - OBJECTIVES: This study was done to highlight the pattern and distribution of trauma deaths in a Nigerian teaching hospital in order to enhance trauma research, improve treatment strategies and prevent trauma deaths. PATIENTS AND METHODS: a prospective data collection was done for 24 months beginning September 1999 detailing the age, sex, occupation, diagnosis, mechanism of trauma, injury arrival time, and circumstances of death and determining retrospectively the severity of trauma using Revised Trauma Score (RTS), Injury Severity Score (ISS) and Trauma and Injury Severity Score (TRISS). RESULTS: There were 129 deaths but 84 (65.1%) had sufficient data for trauma scoring. Male:Female ratio was 60:24 = 2.5:1. The age range was 2.95 years, mean 36.8 +/- 15.2 years for males and 45.5 +/- 23.0 years for females. Two thirds (66.7%) of the deaths occurred among traders/business 27.4%, artisans 20.2%, drivers and students 9.5% each. Three quarters of the deaths (75.0%, 63) were in patients involved in road traffic accidents (RTA) followed by violent trauma (10.7%), falls 9.5% and burns 4.8%. The trauma deaths include head injury (26 deaths, 31.0%), multiple injuries (30.0%), fractures (13.1%), cervical spine injury (10.7), gunshot injuries (8.3%), burns (4.8%) and others (5.0%). Sixty-two patients (73.8%) reached the hospital within 6 hours of the injury. The patients were brought by the Police, Good Samaritans or relatives in nearly equal proportions. The mean RTS, ISS and TRISS probability of survival scores were 5.16, 25 and 67% for males but 6.0, 22 and 75% for females, respectively. CONCLUSIONS: The pattern and distribution of trauma revealed a typical trauma death is a male aged below 40 years, who is a trader/businessman involved in RTA or violent trauma. Despite a probability of survival above 60% and majority of the patients getting to hospital within 6 hours, the inadequate A and E care has thrown up possibilities for prevention of trauma death, improving treatment strategies and enhancing trauma research. PMID- 14529234 TI - Blood pressure response to out-patient drug treatment of hypertension in 1973 1993 at Korle-Bu Teaching Hospital, Accra, Ghana. AB - A retrospective audit of the first twelve months of out patient drug treatment of hypertension at the Korle-Bu Teaching Hospital during the period 1973-1993 is reported. A previous study had shown that at least 7 drug regimes were used to treat hypertension at Korle-Bu during the period. The aim of the present study was to compare the effect and efficacy of these antihypertensive drug treatment regimes on blood pressure during the first 12 months of treatment. Result of 155 (47%) case notes, which met the inclusion criteria, are presented. One month of drug treatment of hypertension significantly reduced systolic and diastolic blood pressure by 21.4 +/- 30.5 (p<0.001) and 13.8 +/- 16.5 (p<0.001) mmHg, respectively. This reduction in blood pressure was maintained to the 12th month. At 12 months, systolic and diastolic blood pressures were unchanged in 19% and 28% of patients, respectively, indicating no response to drug treatment. Recommended target blood pressure of < or = 140/90 mm Hg was achieved in only 25.6% of all patients. All drug treatment regimes significantly reduced blood pressure to a similar extent so that any differences were not statistically significant. However, the efficacy of the drug regimes differed significantly (p=0.02). It was greatest in patients treated with monotherapy with either diuretic or reserpine, intermediate with two drug combinations and least with 3 or 4 drugs. The data showed that diuretics were marginally better than reserpine as first line monotherapy. Furthermore, any diuretic based 2-drug regime was equally efficacious although a beta-blocker or methyldopa as second drug seemed favoured by the data. The addition of a third or fourth drug was counter productive as the increased number of drugs did not decrease blood pressure significantly. PMID- 14529235 TI - Inaccuracies in a published paper--what is the accurate Ilorin experience on angular deformities of the knees in children? PMID- 14529236 TI - Epidemiology of infertility: social problems of the infertile couples. AB - Infertility is of public health importance in Nigeria and many other developing nations because of its high prevalence and especially due to its serious social implications. A review of the epidemiology of infertility in Nigeria and other parts of Sub-Saharan Africa is presented and socio-cultural issues including the social impact on couples are discussed. The major cause of infertility in Africa is infection--STDs, post-abortal and puerperal sepsis. Beliefs about causes, and failure of orthodox methods of treatment have led many couples to seek solution from traditional doctors and faith healers without success. Infertility causes marital disharmony, which often leads to divorce. Women are often blamed for the infertility and men engage in polygyny in an attempt to have children. The couple can also suffer stress from the management of the infertility. Adoption is not popular and assisted reproduction has medico-legal implications. Preventive measures are suggested, including counselling at every stage of the management. PMID- 14529237 TI - Shunt-responsive dementing illness in a Nigerian--a case report. AB - Dementia is a growing medico-social problem worldwide because of the rising population of the elderly. About 10-30% of the cases have been found to be treatable or reversible with the improvement in neuro-diagnostic and neuro imaging techniques. These reversible cases should be diagnosed without over investigating the many patients with irreversible disease. We present a case of a reversible dementia due to Normal Pressure Hydrocephalus with dramatic surgical therapeutic response. PMID- 14529238 TI - Mass casualty management: Jos University Teaching Hospital experience. AB - Three experiences of management of mass casualties in Jos University Teaching Hospital were analysed. Classification into minor, moderate and major mass casualties was done using multiple criteria of number of casualties, number of doctors required to contain the situation, number of nurses and paramedical staff, degree of distruption of hospital services and time required to handle the immediate stabilisation of the casualties. The classification recognises a category called "regional disaster" and attempts to enunciate a principle of initiation, mobilisation and co-ordination of management of such disasters among hospitals and human and material resources within the region. It is envisaged that coalescence of "regional disaster preparedness" would from the bedrock of national ambulance services system. PMID- 14529239 TI - A historical overview of combat stress control units of the U.S. Army. AB - Combat stress control units have been deployed to the Gulf War, Somalia, Haiti, Guantanamo Bay, Bosnia, and Kosovo. They have been very flexible and useful mental health tools for commanders in both combat and peacekeeping operations for the past decade. In their operational role they have been effective, but their garrison mission remains unclear. This article summarizes the uses, missions, and lessons learned from the various combat stress control missions around the world. PMID- 14529241 TI - Human immunodeficiency virus social work program at the Walter Reed Army Medical Center: a historical perspective. AB - Human immunodeficiency virus (HIV) infection has become a pandemic concern for many nations. When this disease first presented itself in a global manner in the early 1980s, it was accompanied by fear, denial, misunderstanding, social stigma, and a paucity of available support services. The U.S. Army was becoming increasingly aware of the potential impact HIV could have on the active forces. A tragic event involving the suicide of a young HIV-infected soldier resulted in the development of a comprehensive medical/psychosocial assessment and treatment program for HIV-infected service members and their families at the Walter Reed Army Medical Center. Social work services played an integral role in the development of this program as this profession has done historically in meeting other emerging needs of the military. Special attention is given to the unique psychosocial issues and needs of the HIV-infected service member and the comprehensive and compassionate response of the military medical team with its significant social work contribution. PMID- 14529240 TI - Treatment results for open comminuted distal humerus intra-articuler fractures with Ilizarov circular external fixator. AB - In open, intra-articular distal humerus fracture caused by gunshot injury, full functional recovery is difficult to obtain. Three basic treatment methods are available: minimal internal fixation, open reduction-internal fixation, and external fixation. In Gulhane Military Medical Academy Department of Orthopedics and Traumatology, 19 of 20 cases of gunshot injuries were treated with circular external fixator between the January 1995 and December 2000. Nine (45%) cases were type III-A, eight (40%) were type III-B, and three (15%] were type III-C. Eight (40%) cases were brought to the hospital 6 to 8 hours after the injury and 12 (60%) were in late stage. An amputation was done in one case. Mean follow-up period was 34.3 (14-55) months. Union was achieved in all 19 of the cases, and circular external fixator was taken out in a mean period of 4.6 (3-7) months. In the early treatment group, three (42.9%) were good, three (42.9%) were moderate, and one (14.2%) was unsatisfactory. In the late treatment group, five (41.7%) were good, four (33.3) were moderate, and three (25%) were unsatisfactory. Circular external fixator can be preferred as a treatment alternative in selected cases of distal humerus intra-articular open communited fractures because it protects the soft tissue connections and blood circulation of bone fractured, permits early elbow movements, and allows the patient to return to daily life very early. PMID- 14529242 TI - Skip policy: road to Force Health Protection 2010. AB - The 2010 Force Health Protection Capstone concept envisions a single level of theater hospitalization and a greater reliance on the strategic movement of casualties from the theater. A significant Force Health Protection implication is 100% of the combat zone patients leaving theater will not have a second stay at an echelon/level IV hospital. In 2000, the Army began moving toward the Force Health Protection concept by using a skip policy for determining 2007 medical force structure requirements. Implementing the skip policy avoids (eliminates) the second echelon/level IV hospital length of stay for a percentage of combat zone patients leaving theater. The Army's decision to implement a skip policy exposed the complexities associated with determining deployable medical force structure requirements and the inherent inter-relatedness of the services medical mission. PMID- 14529243 TI - Pneumococcal vaccine rates in persons age 65 years and older: a U.S. Air Force Medical Facility record review. AB - The pneumococcal vaccine is underused according to self-reported surveys and the full scope of the problem remains undetermined. OBJECTIVE: Primary aims were to determine whether the pneumococcal vaccination rates varied between clinics for patients 65 years of age and older and to compare the results with national survey reports and claims data. METHODS: A total of 368 medical records were examined in this prevalence survey with 268 records from the Internal Medicine Clinic and 100 records from the Family Practice Clinic at Wilford Hall U.S. Air Force Medical Center. RESULTS: Documented vaccination rates were 92.9% and 69.0% for Internal Medicine Clinic and Family Practice Clinic, respectively. This difference in vaccination rates was statistically significant (chi2 = 35.462; df = 1; p < 0.001), and both clinics exceeded rates found in surveys and claims data. CONCLUSIONS: Clinics differ in their vaccination rates and self-reported national survey data may not accurately reflect the prevalence of pneumococcal vaccination among military retirees. PMID- 14529244 TI - Four-year prospective study of military trainees returned to duty following a mental health evaluation. AB - The purpose of the study was to determine the long-term outcome for military trainees referred for a mental health evaluation during basic training. Participants were 261 Air Force basic trainees seen for a mental health evaluation during January through October 1997 and returned to duty. Frequency and type of discharge from the military were documented for participants during a 4-year follow-up period. The results showed that two-thirds of the study sample failed to complete their obligated tour of duty. Most discharges occurred within a 100-day period from the time of the evaluation. Common discharge types were physical condition interfering with performance of duty, entry-level performance and conduct, fraudulent entry, and character or behavior disorder. Implications of the findings for mental health providers working in a military training environment are discussed. PMID- 14529245 TI - Health disturbances of German battle tank officers: results of an interview with 64 commanding officers of a tank battalion. AB - In a former presentation, we reported the failure of commanding officers after a 320-km-long chain march. In this study, 64 commanding officers of a tank battalion were questioned regarding subjective health disturbances similar to the disturbances we have seen before. The soldiers indicated subjective troubles and frequency in dependence of the load duration. After 36 hours, one-half of the soldiers indicated feeling some pain, which consisted primarily of knee and back troubles. In addition to the marked disturbances after 24 hours, 50% of the soldiers indicated suffering from at least one health disturbance, and after 36 hours, 80% of the soldiers suffered at least one health disturbance. The disturbances appear faster and more frequently the longer the soldiers are on the battle tank as commanding officer. There is also a subjective increase in troubles with the years of service. Our results show that the vibration load on the battle tank can cause health disturbances. These disturbances seem to be a relevant problem of the commanding officers with no single case report. PMID- 14529246 TI - Infectious mononucleosis presenting as spontaneous splenic rupture without other symptoms. AB - Splenic rupture is an uncommon complication of infectious mononucleosis (IM), occurring in 0.1% to 0.5% of all patients. It remains the most common lethal complication of IM. Rupture of the spleen with no other symptoms of IM is almost unheard of. This is the report of a case of spontaneous splenic rupture requiring splenectomy in a patient with a positive heterophil antibody test and no other signs or symptoms of IM. The diagnosis and management of splenic rupture in IM are discussed. PMID- 14529247 TI - Gastric bypass: it pays to keep military hospitals busy. AB - This study was performed to review the surgical treatment of obesity at a community military hospital and compare costs to TRICARE reimbursement rates (the cost of sending a patient to a civilian surgeon for obesity surgery). The preoperative, operative, and postoperative phases are described in detail. The expenses of five consecutive patients were calculated and averaged. Each operation at our hospital cost 1,710 dollars, whereas the TRICARE cost was at least 6,950 dollars. A saving of 5,240 dollars per operation was achieved in our military hospital. These five patients subjectively graded their outcomes as very good to excellent at a mean of 7.9 months from surgery. These patients lost an average of 70% of their excess body weight. All patients with weight-related comorbidities reported resolution of at least one problem. PMID- 14529248 TI - Designing a Medical Humanitarian Assistance Course for advanced practice nurses in the uniformed services. AB - The U.S. uniformed services frequently respond to natural and man-made disasters worldwide. Disaster management and humanitarian assistance has been a major focus of military medicine for well over a decade. Training for these missions is a priority for the U.S. military, specifically as a result of the Gulf War. The Medical Humanitarian Assistance Course for advanced practice nurses is a course available to train advanced practice nurses in disaster management and humanitarian assistance. This article will describe the evolution, design, format, and implementation of the Medical Humanitarian Assistance Course. PMID- 14529249 TI - Femoral vessel injuries in modern warfare since Vietnam. AB - The incidence of lower extremity injuries is high in modern warfare; however the mortality rate from these injuries is low. Despite the overall low mortality of lower extremity injuries, many deaths in modern conflicts are due to femoral vessel injury. The modern warfare literature was reviewed. In Somalia, 1 of the 14 reported deaths was due to a laceration of the superficial femoral artery. In the Persian Gulf, three deaths were reported in an Army field hospital, one resulted from uncontrolled hemorrhage from a profunda femoris artery wound and two others resulted from traumatic amputations with consequent major arterial injury. Despite the advances in modern body armor, the groin is left unprotected. A strategy to help minimize U.S. casualties in modern warfare may be a method of field hemostasis specifically designed for femoral vessel injury. PMID- 14529250 TI - Psychosocial and health correlates of types of traumatic event exposures among U.S. military personnel. AB - The prevalence of lifetime exposure to violence, natural disaster, or major accidents involving injuries or fatalities was examined in the largest population based epidemiologic survey of U.S. military personnel to date. The psychosocial and health effects of types of exposure experience (witness only, victim/survivor, relief worker), gender differences, and social support were also evaluated. Over 15,000 active duty U.S. military personnel from stratified random samples of active duty U.S. personnel from all services responded to either mail questionnaires and/or worksite surveys. The lifetime exposure to one or more traumatic events was 65%; the most prevalent trauma for men was witnessing a major accident, and for women, witnessing a natural disaster. Victims of any traumatic event were at twice the risk of having two or more physical and mental health problems than nonexposed controls. Health outcomes of trauma exposure vary by type of traumatic event: type of exposure experience, rank, and gender. PMID- 14529251 TI - Patient satisfaction in military medicine: status and an empirical test of a model. AB - The Department of Defense (DoD) is concerned about how well military medical treatment facilities in the military health system perform. Patient expectations, attitudes, and health care use have been examined in numerous studies; the results are fairly consistent. Eligible beneficiaries report moderate satisfaction with the health care received. In 1994-2001, annual DoD and monthly ambulatory patient surveys were conducted in military medical treatment facilities. The DoD surveys document how patients perceive the care provided. The obvious research concerns are: requirements for conducting surveys; who should be surveyed: eligible beneficiaries or actual users; when; where; representative sample; how often to conduct assessment; data collection methods; analytic schemes; overall trends; predictors of satisfaction; use of results; and timeliness of findings. This study examines these issues and analyzes raw data from selected annual DoD and monthly ambulatory surveys. The overall level of perceived satisfaction has been "good" over the years surveys were used. The model demonstrated the use of examining demographic and attitudinal components of patient satisfaction in military medical facilities. PMID- 14529252 TI - Aerobic capacity of Gulf War veterans with chronic fatigue syndrome. AB - A large overlap exists between the diagnosis of chronic fatigue syndrome (CFS) and the unexplained symptoms reported by many Gulf War veterans (GV). Previous investigations have reported reduced aerobic capacity in civilians with CFS. The present investigation examined metabolic responses to maximal exercise in GVs with CFS compared with healthy GVs. Cardiorespiratory and metabolic responses were recorded during a maximal exercise test on a cycle ergometer. The groups were not different in any demographic category (p > 0.05) or self-reported physical activity (p > 0.05). No differences were observed between groups for maximal oxygen uptake (28.9 +/- 6.7 mL/kg/min for CFS vs. 30.8 +/- 7.1 mL/kg/min for controls; p = 0.39), heart rate (155.8 +/- 16.1 bpm for CFS vs. 163.3 +/- 14.9 bpm for controls; p = 0.17), exercise time (9.6 +/- 1.5 minutes for CFS vs. 10.2 +/- 1.4 minutes for controls; p = 0.26), or workload achieved (208 +/- 36.7 W for CFS vs. 224 +/- 42.9 W for controls; p = 0.25). Likewise, no differences were observed at submaximal intensities (p > 0.05). Compared with healthy controls, GVs who report multiple medically unexplained symptoms and meet criteria for CFS do not show a decreased exercise capacity. Thus, it does not appear that the pathology of the GVs with CFS includes a deficiency with mobilizing the cardiopulmonary system for strenuous physical effort. PMID- 14529253 TI - Potential health hazards of cluster bombing in the Shomali Valley, Afghanistan in October-November 2001. AB - A cluster bomb consists of a canister that opens in mid-air releasing a number of bomblets, which spread over a large area of ground and explode on impact. The exact proportion of bomblets that fail to explode on impact is a matter for debate between military and humanitarian agencies. After the recent bombing of Afghanistan, nine cluster bomb sites in the Shomali Valley were assessed. A total of 317 of 1,818 cluster bomblets had failed to explode (17.4%). Of these 317, 107 (33.7%) had become buried in the ground. Fifty-seven percent of the buried cluster bomblets were embedded deeper than 10 cm. To detect cluster bomblets buried at this depth, large loop metal detectors are needed. PMID- 14529254 TI - Effects of FiO2 on O2 consumption and cardiovascular and hormonal responses to hemorrhage in the goat. AB - This study was conducted to determine whether 100% FiO2 during hemorrhage would improve maintenance of mean arterial blood pressure and to assess the cardiovascular, arginine vasopressin, and renin-angiotensin system roles in the response. This also allowed evaluation of FiO2 effects on the baroreceptor control of these hormone systems. Six conscious female goats were hemorrhaged (0.5 mL x kg(-1) x min(-1)) for 30 minutes, while breathing 11%, 21%, or 100% O2. Mean arterial blood pressure was maintained only when the goats breathed 100% O2. FiO2 did not affect O2 consumption or delivery during hemorrhage. Increases in renin and arginine vasopressin were equal or reduced with 100% FiO2 and therefore did not contribute to the improved mean arterial blood pressure maintenance. Inspiration of 100% FiO2 during hemorrhage, therefore, may improve perfusion pressure with no changes in O2 consumption or delivery. Also, 100% FiO2 eliminated the negative correlation between both hormones and right atrial pressure, suggesting altered baroreceptor control of the hormones. PMID- 14529255 TI - A simple cost-benefit analysis of brief interventions on substance abuse at Naval Medical Center Portsmouth. AB - To determine the impact of brief interventions on substance abuse at the Naval Medical Center Portsmouth, a retrospective review of all admissions in fiscal year 2001 was conducted. Patients receiving brief interventions had significantly lower readmission rates (12.6%) than those not receiving interventions (29.4%). For Internal Medicine patients, this difference was most pronounced: 15.4% as opposed to 40.0%. The average cost of a second admission was 17,834.31 dollars overall but 23,690.78 dollars for Internal Medicine specifically. The lower readmission rate associated with brief interventions represents a benefit of 606,366.54 dollars saved at a cost of 31,508.50 dollars for a cost-benefit ratio of 19:1. The data indicate a cost avoidance opportunity of 713,372.40 dollars if all identified substance abuse patients received interventions. Perhaps most significant, the gap between expected and identified substance use disorders (3.7% vs. 25%) suggests 3,400 unidentified persons who could benefit from interventions. This represents an additional cost avoidance opportunity of 10,200,000.00 dollars. PMID- 14529256 TI - Neuropsychological deficits of a U.S. Army pilot following an anoxic event as a function of cardiac arrest. AB - Anoxic encephalopathy occurs as a result of cardiac arrest, respiratory distress, or carbon monoxide poisoning. This is a case report on the neuropsychological deficits of anoxia in an otherwise previously healthy 36-year-old male pilot. The patient was taking an over-the-counter supplement that included an herb called Ma Huang on the day of his cardiac arrest. Ma Huang is reported to potentially present an increased risk of cardiac infarctions and central nervous system dysfunctions. Several instances of death have been linked to Ma Huang. The patient produced a neuropsychological profile that evidenced impairments in executive functioning, memory, language, attention, intellectual and academic functioning, as well as motor speed and coordination, all of which are consistent with diffuse brain damage. This case adds to the body of literature documenting the physical and neuropsychological effects of anoxia, as well as the effects of ephedrine-based supplements, such as Ma Huang. PMID- 14529257 TI - Latex allergy literature review: evidence for making military treatment facilities latex safe. PMID- 14529259 TI - OODA loop. PMID- 14529260 TI - The problems of drugs and addiction. PMID- 14529258 TI - Vaccines and biological substances offered in clinical practice have to be stored all the time in controlled temperatures. PMID- 14529261 TI - Substance use and mental health in Greek warship personnel. PMID- 14529262 TI - Atypical antipsychotics, improved intrusive symptoms in patients with posttraumatic stress disorder. PMID- 14529264 TI - Chicken soup. PMID- 14529263 TI - Back pain, a common problem in primary care. PMID- 14529265 TI - Steroid ring hydroxylation patterns govern cooperativity in human bile acid binding protein. AB - Human ileal bile acid binding protein (I-BABP) is a member of the intracellular lipid binding protein family. This protein is thought to function in the transcellular transport and enterohepatic circulation of bile salts. Human I-BABP binds two molecules of glycocholate, the physiologically most abundant bile salt, with modest intrinsic affinity but a remarkably high degree of positive cooperativity. Here we report a calorimetric analysis for the binding of a broad panel of bile salts to human I-BABP. The interaction of I-BABP with nine physiologically relevant derivatives of cholic acid, chenodeoxycholic acid, and deoxycholic acid in their conjugated (glycine and taurine) and unconjugated forms was monitored by isothermal titration calorimetry. All bile salts bound to I-BABP with a 2:1 stoichiometry and similar overall affinity, but the derivatives of cholic acid displayed much higher Hill coefficients, a measure of macroscopic positive cooperativity. To test whether the cooperativity was dependent on individual structural features of the bile salt side chain, a series of side chain-extended bile salts that lacked a hydrogen bond donor or acceptor at C-24 were chemically synthesized. These synthetic variants exhibited the same energetic and cooperativity profile as the naturally occurring bile salts. Our findings indicate that cooperativity in bile salt-I-BABP recognition is governed by the pattern of steroid B- and C-ring hydroxylation and not the presence or type of side-chain conjugation. PMID- 14529266 TI - Energetics of sequence-specific protein-DNA association: computational analysis of integrase Tn916 binding to its target DNA. AB - The N-terminal domain of the bacterial integrase Tn916 specifically recognizes the 11 bp DNA target site by positioning the face of a three-stranded beta-sheet into the major groove. Binding is linked to structural adaptation. We have characterized INT-DBD binding to DNA in detail by calorimetry [Milev, S., Gorfe, A., Karshikoff, A., Clubb, R. T., Bosshard, H. R., and Jelesarov, I. (2003) Biochemistry 42, 3481-3491]. Our thermodynamic analysis has indicated that the major driving force of association is the hydrophobic effect while polar interactions contribute less. To gain more comprehensive information about the binding process, we performed a computational analysis of the binding free energy and report here the results. A hybrid molecular mechanics/continuum approach was followed. The total binding free energy is predicted with reasonable accuracy. The calculations confirm that nonpolar effects stabilize the protein-DNA complex while electrostatics opposes binding. Structural changes optimizing surface complementarity are costly in terms of energy. The energetic consequences from the replacement of nine DNA-contacting residues by alanine were investigated. The calculations correctly predict the binding affinity decrease of eight mutations and the destabilizing effect of one wild-type residue. Bulky side chains stabilize the wild-type complex through packing interactions and favorable nonpolar dehydration, but the net nonpolar energy changes do not correlate with the relative affinity loss upon mutation. Discrete protein-DNA electrostatic interactions may be net stabilizing or net destabilizing depending on the local environment. In contrast to nonpolar energy changes, the magnitude of the electrostatic free energy ranks the mutations according to the experimentally measured DeltaDeltaG. Free energy decomposition analysis from a structural perspective leads to detailed information about the thermodynamic strategy used by INT-DBD for sequence-specific DNA binding. PMID- 14529267 TI - Biochemical and spectroscopic studies on (S)-2-hydroxypropylphosphonic acid epoxidase: a novel mononuclear non-heme iron enzyme. AB - The last step of the biosynthesis of fosfomycin, a clinically useful antibiotic, is the conversion of (S)-2-hydroxypropylphosphonic acid (HPP) to fosfomycin. Since the ring oxygen in fosfomycin has been shown in earlier feeding experiments to be derived from the hydroxyl group of HPP, this oxirane formation reaction is effectively a dehydrogenation process. To study this unique C-O bond formation step, we have overexpressed and purified the desired HPP epoxidase. Results reported herein provided initial biochemical evidence revealing that HPP epoxidase is an iron-dependent enzyme and that both NAD(P)H and a flavin or flavoprotein reductase are required for its activity. The 2 K EPR spectrum of oxidized iron-reconstituted fosfomycin epoxidase reveals resonances typical of S = (5)/(2) Fe(III) centers in at least two environments. Addition of HPP causes a redistribution with the appearance of at least two additional species, showing that the iron environment is perturbed. Exposure of this sample to NO elicits no changes, showing that the iron is nearly all in the Fe(III) state. However, addition of NO to the Fe(II) reconstituted enzyme that has not been exposed to O(2) yields an intense EPR spectrum typical of an S = (3)/(2) Fe(II)-NO complex. This complex is also heterogeneous, but addition of substrate converts it to a single, homogeneous S = (3)/(2) species with a new EPR spectrum, suggesting that substrate binds to or near the iron, thereby organizing the center. The fact that NO binds to the ferrous center suggests O(2) can also bind at this site as part of the catalytic cycle. Using purified epoxidase and (18)O isotopic labeled HPP, the retention of the hydroxyl oxygen of HPP in fosfomycin was demonstrated. While ether ring formation as a result of dehydrogenation of a secondary alcohol has precedence in the literature, these catalyses require alpha-ketoglutarate for activity. In contrast, HPP epoxidase is alpha-ketoglutarate independent. Thus, the cyclization of HPP to fosfomycin clearly represents an intriguing conversion beyond the scope entailed by common biological epoxidation and C-O bond formation. PMID- 14529268 TI - Inhibition of dog and human gastric lipases by enantiomeric phosphonate inhibitors: a structure-activity study. AB - The crystal structures of gastric lipases in the apo form [Roussel, A., et al. (1999) J. Biol. Chem. 274, 16995-17002] or in complex with the (R(P))-undecyl butyl phosphonate [C(11)Y(4)(+)] [Roussel, A., et al. (2002) J. Biol. Chem. 277, 2266-2274] have improved our understanding of the structure-activity relationships of acid lipases. In this report, we have performed a kinetic study with dog and human gastric lipases (DGL and HGL, respectively) using several phosphonate inhibitors by varying the absolute configuration of the phosphorus atom and the chain length of the alkyl/alkoxy substituents. Using the two previously determined structures and that of a new crystal structure obtained with the other (S(P))-phosphonate enantiomer [C(11)Y(4)(-)], we constructed models of phosphonate inhibitors fitting into the active site crevices of DGL and HGL. All inhibitors with a chain length of fewer than 12 carbon atoms were found to be completely buried in the catalytic crevice, whereas longer alkyl/alkoxy chains were found to point out of the cavity. The main stereospecific determinant explaining the stronger inhibition of the S(P) enantiomers is the presence of a hydrogen bond involving the catalytic histidine as found in the DGL-C(11)Y(4)(-) complex. On the basis of these results, we have built a model of the first tetrahedral intermediate corresponding to the tristearoyl-lipase complex. The triglyceride molecule completely fills the active site crevice of DGL, in contrast with what is observed with other lipases such as pancreatic lipases which have a shallower and narrower active site. For substrate hydrolysis, the supply of water molecules to the active site might be achieved through a lateral channel identified in the protein core. PMID- 14529270 TI - Relaxing the nicotinamide cofactor specificity of phosphite dehydrogenase by rational design. AB - Homology modeling was used to identify two particular residues, Glu175 and Ala176, in Pseudomonas stutzeri phosphite dehydrogenase (PTDH) as the principal determinants of nicotinamide cofactor (NAD(+) and NADP(+)) specificity. Replacement of these two residues by site-directed mutagenesis with Ala175 and Arg176 both separately and in combination resulted in PTDH mutants with relaxed cofactor specificity. All three mutants exhibited significantly better catalytic efficiency for both cofactors, with the best kinetic parameters displayed by the double mutant, which had a 3.6-fold higher catalytic efficiency for NAD(+) and a 1000-fold higher efficiency for NADP(+). The cofactor specificity was changed from 100-fold in favor of NAD(+) for the wild-type enzyme to 3-fold in favor of NADP(+) for the double mutant. Isoelectric focusing of the proteins in a nondenaturing gel showed that the replacement with more basic residues indeed changed the effective pI of the protein. HPLC analysis of the enzymatic products of the double mutant verified that the reaction proceeded to completion using either substrate and produced only the corresponding reduced cofactor and phosphate. Thermal inactivation studies showed that the double mutant was protected from thermal inactivation by both cofactors, while the wild-type enzyme was protected by only NAD(+). The combined results provide clear evidence that Glu175 and Ala176 are both critical for nicotinamide cofactor specificity. The rationally designed double mutant might be useful for the development of an efficient in vitro NAD(P)H regeneration system for reductive biocatalysis. PMID- 14529269 TI - Determination of the rate of reduction of oxyferrous cytochrome P450 2B4 by 5 deazariboflavin adenine dinucleotide T491V cytochrome P450 reductase. AB - The use of 5-deazaFAD T491V cytochrome P450 reductase has made it possible to directly measure the rate of electron transfer to microsomal oxyferrous cytochrome (cyt) P450 2B4. In this reductase the FMN moiety can be reduced to the hydroquinone, FMNH(2), while the 5-deazaFAD moiety remains oxidized [Zhang, H., et al. (2003) Biochemistry 42, 6804-6813]. The rate of electron transfer from 5 deazaFAD cyt P450 reductase to oxyferrous cyt P450 was determined by rapidly mixing the ferrous cyt P450-2-electron-reduced 5-deazaFAD T491V reductase complex with oxygen in the presence of substrate. The 5-deazaFAD T491V reductase which can only donate a single electron reduces the oxyferrous cyt P450 and oxidizes to the air-stable semiquinone, with rate constants of 8.4 and 0.37 s(-1) at 15 degrees C. Surprisingly, oxyferrous cyt P450 turns over more slowly with a rate constant of 0.09 s(-1), which is the rate of catalysis under steady-state conditions at 15 degrees C (k(cat) = 0.08 s(-1)). In contrast, the rate constant for electron transfer from ferrous cyt b(5) to oxyferrous cyt P450 is 10 s(-1) with oxyferrous cyt P450 and cyt b(5) simultaneously undergoing spectral changes. Quantitative analyses by LC-MS/MS revealed that the product, norbenzphetamine, was formed with a coupling efficiency of 52% with cyt b(5) and 32% with 5 deazaFAD T491V reductase. Collectively, these results suggest that during catalysis a relatively stable reduced oxyferrous intermediate of cyt P450 is formed in the presence of cyt P450 reductase but not cyt b(5) and that the rate limiting step in catalysis follows introduction of the second electron. PMID- 14529271 TI - Site-directed sulfhydryl labeling of the oxaloacetate decarboxylase Na+ pump of Klebsiella pneumoniae: helix VIII comprises a portion of the sodium ion channel. AB - Helix VIII of the beta-subunit of the oxaloacetate decarboxylase of Klebsiella pneumoniae contains the functionally important residues betaN373, betaG377, betaS382, and betaR389. Using a functional oxaloacetate decarboxylase mutant devoid of Cys residues in the beta-subunit, each amino acid residue in helix VIII was replaced individually with Cys. Structural and dynamic features of this region were studied by using site-directed sulfhydryl modification of 20 single Cys replacement mutants with methanethiosulfonate (MTS) reagents in the absence or presence of Na(+) ions. The pattern of accessibility of the MTS reagents from the periplasmic side of helix VIII shows a periodicity which suggests that this region is alpha-helical. In particular, a water-accessible face comprising betaN373, betaG377, betaS382, betaM386, and betaV390 may be part of a Na(+) channel. Cys residues introduced in the cytoplasmically oriented part of helix VIII were accessible to three different water-soluble MTS compounds and therefore believed to be exposed to water on this side of the membrane. Most residues located in the upper part of helix VIII (residues betaN373-betaV381C) were protected by Na(+) ions for inactivation by the MTS reagents. The distinct results on accessibility toward the different MTS reagents obtained in the presence or absence of Na(+) ions may suggest a conformational change upon binding of Na(+) in this region. The betaR389C mutant had a reduced activity and a pH optimum at pH 9, which could be restored to a wild-type pH optimum of 6.5 and to a 400% gain in activity upon chemical modification with 2-aminoethyl methanethiosulfonate. PMID- 14529272 TI - Rice C2-domain proteins are induced and translocated to the plasma membrane in response to a fungal elicitor. AB - Hundreds of proteins involved in signaling pathways contain a Ca(2+)-dependent membrane-binding motif called the C2-domain. However, no small C2-domain proteins consisting of a single C2-domain have been reported in animal cells. We have isolated two cDNA clones, OsERG1a and OsERG1b, that encode two small C2-domain proteins of 156 and 159 amino acids, respectively, from a fungal elicitor-treated rice cDNA library. The clones are believed to have originated from a single gene by alternative splicing. Transcript levels of the OsERG1 gene are dramatically elevated by a fungal elicitor prepared from Magnaporthe grisea or by Ca(2+) ions. The OsERG1 protein produced in Escherichia coli binds to phospholipid vesicles in a Ca(2+)-dependent manner and is translocated to the plasma membrane of plant cells by treatment with either a fungal elicitor or a Ca(2+) ionophore. These results suggest that OsERG1 proteins containing a single C2-domain are involved in plant defense signaling systems. PMID- 14529273 TI - Insights into a single rod-like helix in activated radixin required for membrane cytoskeletal cross-linking. AB - The members of the ezrin-radixin-moesin (ERM) family of proteins function as membrane-cytoskeletal cross-linkers in actin-rich cell surface structures. ERM proteins are thereby thought to be essential for cortical cytoskeleton organization, cell motility, adhesion, and proliferation. These modular polypeptides consist of a central helix-rich region, termed the alpha-domain, that connects an N-terminal FERM domain required for membrane binding and a C terminal region which contains a major actin-binding motif. Conformational regulation of ERM protein function occurs by association of the FERM and C terminal domains, whereby the membrane- and actin-binding activities are mutually suppressed and the protein is thought to take an inactive "closed" form. Here we report in vitro and in vivo studies of radixin to address the role of the alpha domain in conformational activation of ERM proteins. Remarkably, an isolated alpha-domain comprised of radixin(311-469) forms a monomeric, stable helical rod that spans 240 A in length from the N-terminus to the C-terminus, most likely stabilized by extensive salt bridge interactions. By fusing green fluorescent protein variants to the FERM and C-terminal domains, we probed in vitroconformational changes impacted by the presence of the alpha-domain using fluorescence resonance energy transfer (FRET). Furthermore, deletion of this unusually long alpha-helical structure (radixin residues 314-411) prevents ERM membrane targeting in vivo. PMID- 14529274 TI - Protonation structures of Cys-sulfinic and Cys-sulfenic acids in the photosensitive nitrile hydratase revealed by Fourier transform infrared spectroscopy. AB - Nitrile hydratase (NHase) from Rhodococcus N-771, which catalyzes hydration of nitriles to the corresponding amides, exhibits novel photosensitivity; in the dark, it is in the inactive form that binds an endogenous nitric oxide (NO) molecule at the non-heme iron center, and photodissociation of the NO activates the enzyme. NHase is also known to have a unique active site structure. Two cysteine ligands to the iron center, alphaCys112 and alphaCys114, are post translationally modified to sulfinic acid (Cys-SO(2)H) and sulfenic acid (Cys SOH), respectively, which are thought to play a crucial role in the catalytic reaction. Here, we have determined the protonation structures of these Cys-SO(2)H and Cys-SOH groups using Fourier transform infrared (FTIR) spectroscopy in combination with density functional theory (DFT) calculations. The light-induced FTIR difference spectrum of NHase between the dark inactive and light active forms exhibited two prominent signals at (1154-1148)/1126 and (1040-1034)/1019 cm(-1), which downshifted to 1141/1114 and 1026/1012 cm(-1), respectively, in the uniformly (34)S-labeled NHase. In addition, a minor signal at 915/908 cm(-1) also showed a considerable downshift upon (34)S labeling. These (34)S-sensitive signals were basically conserved in D(2)O buffer with only slight shifts. Vibrational frequencies of methanesulfenic acid (CH(3)SOH) and methanesulfinic acid (CH(3)SO(2)H), simple model compounds of Cys-SOH and Cys-SO(2)H, respectively, were calculated using the DFT method in both the protonated and deprotonated forms and in metal complexes. Comparison of the calculated frequencies and isotope shifts with the observed ones provided the assignment of the two major signals around 1140 and 1030 cm(-1) to the asymmetric and symmetric SO(2) stretching vibrations, respectively, of the S-bonded Cys-SO(2)(-) complex, and the assignment of the minor signal around 910 cm(-1) most likely to the SO stretch of the S-bonded Cys-SO(-) complex. These assignments and the small frequency shifts upon deuteration are consistent with the view that the deprotonated alphaCys112-SO(2)(-) and alphaCys114-SO(-) are hydrogen-bonded with the protons from betaArg56 and/or betaArg141, forming a reactive cavity at the interface of the alpha and beta subunits. There is further speculation that either of these groups is hydrogen bonded to a reactant water molecule, increasing its basicity to facilitate the nucleophilic attack on the nitrile substrate bound to the iron center. PMID- 14529275 TI - Continuum electrostatic model for the binding of cytochrome c2 to the photosynthetic reaction center from Rhodobacter sphaeroides. AB - Electrostatic interactions are important for protein-protein association. In this study, we examined the electrostatic interactions between two proteins, cytochrome c(2) (cyt c(2)) and the reaction center (RC) from the photosynthetic bacterium Rhodobacter sphaeroides, that function in intermolecular electron transfer in photosynthesis. Electrostatic contributions to the binding energy for the cyt c(2)-RC complex were calculated using continuum electrostatic methods based on the recent cocrystal structure [Axelrod, H. L., et al. (2002) J. Mol. Biol. 319, 501-515]. Calculated changes in binding energy due to mutations of charged interface residues agreed with experimental results for a protein dielectric constant epsilon(in) of 10. However, the electrostatic contribution to the binding energy for the complex was close to zero due to unfavorable desolvation energies that compensate for the favorable Coulomb attraction. The electrostatic energy calculated as a function of displacement of the cyt c(2) from the bound position showed a shallow minimum at a position near but displaced from the cocrystal configuration. These results show that although electrostatic steering is present, other short-range interactions must be present to contribute to the binding energy and to determine the structure of the complex. Calculations made to model the experimental data on association rates indicate a solvent separated transition state for binding in which the cyt c(2) is displaced approximately 8 A above its position in the bound complex. These results are consistent with a two-step model for protein association: electrostatic docking of the cyt c(2) followed by desolvation to form short-range van der Waals contacts for rapid electron transfer. PMID- 14529276 TI - Characterization of the membrane binding mode of the C2 domain of PKC epsilon. AB - PKCepsilon is a member of the group of novel PKCs that contain a C2 domain located in their N-terminal region. On the basis of recent structural studies, a series of mutants were prepared to increase our knowledge of the mechanism of the phospholipid binding site of this domain. The results revealed that this domain preferentially binds to phosphatidic acid- and phosphatidylserine-containing vesicles. Although the increase in affinity was linear in the case of phosphatidic acid, it became exponential when the vesicles contained increasing concentrations of phosphatidylserine. Site-directed mutagenesis studies showed that residues W23, R26, and R32 located in loop 1 and I89 and Y91 located in loop 3 are of critical importance when the binding is performed with phosphatidic acid containing vesicles. Furthermore, when the same mutants were assayed with phosphatidylserine-containing vesicles, no binding was observed in any case, reflecting the smaller affinity of the C2 domain for phosphatidylserine containing vesicles. A study of the ionic nature of the membrane interaction suggested that it is mainly driven by electrostatic interactions that are disrupted by very low salt concentrations. Differential scanning calorimetry experiments performed to ascertain whether this interaction affected the transition phase of the phosphatidic acid demonstrated that increasing concentrations of the protein lead to changes in the transition, with more than one peak appearing at lower temperatures, which suggests a weak interaction focused on the polar headgroup of the phospholids. In conclusion, a different membrane-binding mode from those previously described in other C2 domains has been found and is seemingly based on electrostatic, interfacial, and hydrophobic interactions without the participation of Ca(2+) ions. PMID- 14529277 TI - Structural study of the C2 domains of the classical PKC isoenzymes using infrared spectroscopy and two-dimensional infrared correlation spectroscopy. AB - The secondary structure of the C2 domains of the classical PKC isoenzymes, alpha, betaII, and gamma, has been studied using infrared spectroscopy. Ca(2+) and phospholipids were used as protein ligands to study their differential effects on the isoenzymes and their influence on thermal protein denaturation. Whereas the structures of the three isoenzymes were similar in the absence of Ca(2+) and phospholipids at 25 degrees C, some differences were found upon heating in their presence, the C2 domain of the gamma-isoenzyme being better preserved from thermal denaturation than the domain from the alpha-isoenzyme and this, in turn, being better than that from the beta-isoenzyme. A two-dimensional correlation study of the denaturation of the three domains also showed differences between them. Synchronous 2D-IR correlation showed changes (increased aggregation of denaturated protein) occurring at 1616-19 cm(-1), and this was found in the three isoenzymes. On the other hand, the asynchronous 2D-IR correlation study of the domains in the absence of Ca(2+) showed that, in all cases, the aggregation of denaturated protein increased after changes in other structural components, an increase perhaps related with the hard-core role of the beta-sandwich in these proteins. The differences observed between the three C2 domains may be related with their physiological specialization and occurrence in different cell compartments and in different cells. PMID- 14529278 TI - Humoral immune response to fibrillar beta-amyloid peptide. AB - The beta-amyloid peptide (Abeta) is a normal product of the proteolytic processing of its precursor (beta-APP). Normally, it elicits a very low humoral immune response; however, the aggregation of monomeric Abeta to form fibrillar Abeta amyloid creates a neo-epitope, to which antibodies are generated. Rabbits were injected with fibrillar human Abeta(1-42), and the resultant antibodies were purified and their binding properties characterized. The antibodies bound to an epitope in the first eight residues of Abeta and required a free amino terminus. Additional residues did not affect the affinity of the epitope as long as the peptide was unaggregated; the antibody bound Abeta residues 1-8, 1-11, 1-16, 1 28, 1-40, and 1-42 with similar affinities. In contrast, the antibodies bound approximately 1000-fold more tightly to fibrillar Abeta(1-42). Their enhanced affinity did not result from their bivalent nature: monovalent Fab fragments exhibited a similar affinity for the fibrils. Nor did it result from the particulate nature of the epitope: monomeric Abeta(1-16) immobilized on agarose and soluble Abeta(1-16) exhibited similar affinities for the antifibrillar antibodies. In addition, antibodies raised to four nonfibrillar peptides corresponding to internal Abeta sequences did not exhibit enhanced affinity for fibrillar Abeta(1-42). Antibodies directed to the C-terminus of Abeta bound poorly to fibrillar Abeta(1-42), which is consistent with models where the carboxyl terminus is buried in the interior of the fibril and the amino terminus is on the surface. When used as an immunohistochemical probe, the antifibrillar Abeta(1-42) IgG exhibited enhanced affinity for amyloid deposits in the cerebrovasculature. We hypothesize either that the antibodies recognize a specific conformation of the eight amino-terminal residues of Abeta, which is at least 1000-fold more favored in the fibril than in monomeric peptides, or that affinity maturation of the antibodies produces an additional binding site for the amino-terminal residues of an adjacent Abeta monomer. In vivo this specificity would direct the antibody primarily to fibrillar vascular amyloid deposits even in the presence of a large excess of monomeric Abeta or its precursor. This observation may explain the vascular meningeal inflammation that developed in Alzheimer's disease patients immunized with fibrillar Abeta. Passive immunization with an antibody directed to an epitope hidden in fibrillar Abeta and in the transmembrane region of APP might be a better choice in the search for an intervention to remove Abeta monomers without provoking an inflammatory response. PMID- 14529279 TI - Characterization of the specificity of arginine-specific gingipains from Porphyromonas gingivalis reveals active site differences between different forms of the enzymes. AB - Porphyromonas gingivalis is a pathogen associated with periodontal disease, and arginine-specific proteases (gingipains-R) from the bacterium are important virulence factors. The specificity of two forms of gingipain-R, HRgpA and RgpB, for substrate positions C-terminal to the cleavage site was analyzed, and notable differences were observed between the enzymes. Molecular modeling of the HRgpA catalytic domain, based on the structure of RgpB, revealed that there are four amino acid substitutions around the active site of HRgpA relative to RgpB that may explain their different specificity. Previously, differences in the ability of these two gingipain-R forms to cleave a number of proteins were attributed to additional adhesins on HRgpA mediating increased interaction with the substrates. Here, purified RgpA(cat), the catalytic domain of HRgpA, which like RgpB also lacks adhesin subunits, was used to show that the differences between HRgpA and RgpB are probably due to the amino acid substitutions at the active site. The kinetics of cleavage of fibrinogen, a typical protein substrate for the gingipain R enzymes, which is bound by HRgpA but not RgpA(cat) or RgpB, were evaluated, and it was shown that there was no difference in the cleavage of the fibrinogen Aalpha-chain between the different enzyme forms. HRgpA degraded the fibrinogen Bbeta-chain more efficiently, generating distinct cleavage products. This indicates that while the adhesin domain(s) play(s) a minor role in the cleavage of protein substrates, the major effect is still provided by the amino acid substitutions at the active site of rgpA gene products versus those of the rgpB gene. PMID- 14529280 TI - Design of specific peptide inhibitors for group I phospholipase A2: structure of a complex formed between phospholipase A2 from Naja naja sagittifera (group I) and a designed peptide inhibitor Val-Ala-Phe-Arg-Ser (VAFRS) at 1.9 A resolution reveals unique features. AB - Phospholipase A(2) (PLA(2)) (E. C. 3.1.1.4) is a common enzyme in the two-way cascade mechanism leading to the production of proinflammatory compounds known as eicosanoids. The binding of phospholipase A(2) to the membrane surface and hydrolysis of phospholipids are thought to involve the formation of a hydrophobic channel into which a single substrate molecule diffuses before its cleavage. To regulate the production of proinflammatory compounds, a specific peptide inhibitor Val-Ala-Phe-Arg-Ser (VAFRS) for the group I PLA(2) enzymes has been designed and synthesized. PLA(2) was isolated from Indian cobra (Naja naja sagittifera) venom and purified to homogeneity. The binding studies indicated the K(i) value of 1.02 +/- 0.10 x 10(-8) M. The purified PLA(2) samples and the designed inhibitor VAFRS were cocrystallized. The crystal structure of the complex was determined and refined to 1.9 A resolution. The peptide binds to PLA(2) at the active site and fills the hydrophobic channel completely. However, its placement with respect to the channel is in the opposite direction as compared to those observed in group II PLA(2)'s. Furthermore, the predominant intermolecular interactions involve strong electrostatic interactions between the side chains of peptide Arg and Asp 49 of PLA(2) together with a number of van der Waals interactions with other residues. A good number of observed interactions between the peptide and the protein indicate the significance of a structure based drug design approach. The novel factor in the present sequence of the peptide is related to the introduction of a positively charged residue at the C terminal part of the peptide. PMID- 14529281 TI - Crystal structure of sulerythrin, a rubrerythrin-like protein from a strictly aerobic archaeon, Sulfolobus tokodaii strain 7, shows unexpected domain swapping. AB - Sulerythrin is the first rubrerythrin-like protein to be isolated from an aerobic organism, Sulfolobus tokodaii strain 7, and it lacks a C-terminal rubredoxin-like FeS(4) domain. The protein purified from Sulfolobus cells was crystallized, and the crystal structure was determined at 1.7 A resolution. The dimer of sulerythrin exhibited "domain-swapping" at the loop connecting alphaB and alphaC, hybrid four-helix bundles consisting of alphaA/B and alphaC/D being formed. The structure and atomic identity of the binuclear metal center were determined by means of anomalous scattering analysis. The site contained 1.0 mol of hexacoordinate Fe, 0.80-0.87 mol of tetracoordinate Zn, and 0.73-0.88 mol of putative O(2) per monomer. The metal ions were found at exchanged positions compared to those in the Fe/Zn-containing rubrerythrin from Desulfovibrio vulgaris. The results demonstrate that the binuclear metal center of rubrerythrin like proteins is plastic in its ability to bind metal ions. PMID- 14529282 TI - Receptor-regulated dynamic interaction between endothelial nitric oxide synthase and calmodulin revealed by fluorescence resonance energy transfer in living cells. AB - The endothelial isoform of nitric oxide synthase (eNOS), a key regulator of vascular tone, is activated in endothelial cells by diverse Ca(2+)-mobilizing agonists, including vascular endothelial growth factor (VEGF). Although the activation state of eNOS and the subcellular localization of the enzyme are both highly regulated, the relationship between enzyme activity and subcellular targeting remains obscure. We aim here to elucidate this relationship by direct dynamic imaging analysis of Ca(2+)/CaM-dependent eNOS activation in living endothelial cells, using high-resolution confocal microscopy and donor dequenching fluorescence resonance energy transfer (FRET) techniques. Confocal images show a complex pattern of eNOS subcellular distribution; the enzyme is concentrated in both the plasma membrane and internal membranes, with robust expression in the perinuclear region. We construct a fusion protein between eNOS and the FRET-based calcium sensor cameleon, and analyze the temporal and spatial pattern of VEGF-mediated calcium mobilization using donor dequenching FRET methods. We find that VEGF promotes rapid mobilization of intracellular calcium throughout the regions of the cell in which eNOS is distributed. We further create a series of fusion proteins and use FRET imaging methods to study the interactions between eNOS and its obligate allosteric activator protein calmodulin. We clone the FRET acceptor EYFP (enhanced yellow fluorescent protein) at the C-terminus of calmodulin, and the FRET donor ECFP (enhanced cyan fluorescent protein) into eNOS at a site adjacent to its calmodulin-binding domain. FRET imaging analysis of individual endothelial cells cotransfected with eNOS-ECFP and calmodulin-EYFP shows that VEGF induces interactions between eNOS and calmodulin wherever both are present in the cell. Our studies provide evidence that the pool of rapidly responsive receptor-activated eNOS is distributed throughout endothelial cells in both plasma membrane and internal membrane structures, and that this distribution parallels the localization of agonist-induced intracellular Ca(2+) changes in the vicinity of eNOS. PMID- 14529284 TI - Structural competition involving G-quadruplex DNA and its complement. AB - Structural competition between the G-quadruplex, the I-motif, and the Watson Crick duplex has been implicated for repetitive DNA sequences, but the competitive mechanism of these multistranded structures still needs to be elucidated. We investigated the effects of sequence context, cation species, and pH on duplex formation by the G-quadruplex of dG(3)(T(2)AG(3))(3) and its complement the I-motif of d(C(3)TA(2))(3)C(3), using ITC, DSC, PAGE, CD, UV, and CD stopped-flow kinetic techniques. ITC and PAGE experiments confirmed Watson Crick duplex formation by the complementary strands. The binding constant of the two DNA strands in the presence of 10 mM Mg(2+) at pH 7.0 was shown to be 5.28 x 10(7) M(-1) at 20 degrees C, about 400 times larger than that in the presence of 100 mM Na(+) at pH 5.5. The dynamic transition traces of the duplex formation from the equimolar mixture of G-/C-rich complementary sequences were obtained at both pH 7.0 and pH 5.5. Fitting to a single-exponential function gave an observed rate of 8.06 x 10(-3) s(-1) at 20 degrees C in 10 mM Mg(2+) buffer at pH 7.0, which was about 10 times the observed rate at pH 5.5 under the same conditions. Both of the observed rates increased as temperature rose, implying that the dissociation of the single-stranded structured DNAs is the rate-limiting step for the WC duplex formation. The difference between the apparent activation energy at pH 7.0 and that at pH 5.5 reflects the fact that pH significantly influences the structural competition between the G-quadruplex, the I-motif, and the Watson Crick duplex, which also implies a possible biological role for I-motifs in biological regulation. PMID- 14529283 TI - Bacterial expression, characterization, and disulfide bond determination of soluble human NTPDase6 (CD39L2) nucleotidase: implications for structure and function. AB - The ectonucleoside triphosphate diphosphohydrolases (NTPDases) control extracellular nucleotide concentrations, thereby modulating many important biological responses, including blood clotting and pain perception. NTPDases1-4 are oligomeric integral membrane proteins, whereas NTPDase5 (CD39L4) and NTPDase6 (CD39L2) are soluble monomeric enzymes, making them more amenable to thorough structural and functional analyses than the membrane-bound forms. Therefore, we report here the bacterial expression, refolding, purification, and biochemical characterization of the soluble portion of human NTPDase6. Consistent with the enzyme expressed in mammalian cells, this recombinant NTPDase6 efficiently hydrolyzes GDP, IDP, and UDP (specific activity of approximately 50000 micromol mg(-1) h(-1)), with slower hydrolysis of CDP, ITP, GTP, CTP, ADP, and UTP and virtually no hydrolysis of ATP. The K(m) for GDP (130 +/- 30 microM) is similar to that determined for the soluble rat NTPDase6 expressed in mammalian cells. The secondary structure of the refolded enzyme was determined by circular dichroism to be 33% alpha-helix, 18% beta-sheet, and 49% random coil, consistent with the secondary structure predicted from the amino acid sequence of soluble NTPDase6. Four of the five cysteine residues in the soluble NTPDase6 are highly conserved among all the NTPDases, while the fifth residue is not. Mutation of this nonconserved cysteine resulted in an enzyme very similar to wild type in its enzymology and secondary structure, indicating that this cysteine exists as a free sulfhydryl and is not essential for structure or function. The disulfide pairing of the other four cysteine residues was determined as Cys(249)-Cys(280) and Cys(340)-Cys(354) by HPLC and mass spectral analysis of tryptic peptides. Due to conservation of these cysteine residues, these two disulfide bonds are likely to exist in all NTPDases. A structural model for NTPDase6, incorporating these and other findings obtained with other NTPDases, is proposed. PMID- 14529285 TI - Unified description of electrophoresis and diffusion for DNA and other polyions. AB - The electrophoretic mobilities and diffusion coefficients of single- and double stranded DNA molecules up to 50,000 bases or base pairs in size have been analyzed, using mobilities and diffusion coefficients either measured by capillary electrophoresis or taken from the literature. The Einstein equation suggests that the electrophoretic mobilities (mu) and diffusion coefficients (D) should be related by the expression mu/D = Q/k(B)T, where Q is the charge of the polyion (Q = ze(o), where z is the number of charged residues and e(o) is the fundamental electronic charge), k(B) is Boltzmann's constant, and T is the absolute temperature. If this equation were true, the ratio mu/zD should be a constant equal to e(o)/k(B)T (39.6 V(-1)) at 20 degrees C. However, the ratio mu/zD decreases with an increase in molecular weight for both single- and double stranded DNAs. The mobilities and diffusion coefficients are better described by the modified Einstein equation mu/N(m)D = e(o)/k(B)T, where N is the number of repeat units (bases or base pairs) in the DNA and m is a constant equal to the power law dependence of the diffusion coefficients on molecular weight. The average value of the ratio mu/N(m)D is 40 +/- 4 V(-1) for 36 single- and double stranded DNA molecules of different sizes, close to the theoretically expected value. The generality of the modified Einstein equation is demonstrated by analyzing literature values for sodium polystyrenesulfonate (PSS). The average value of the ratio mu/N(m)D is 35 +/- 6 V(-1) for 14 PSS samples containing up to 855 monomers. PMID- 14529286 TI - DNA sequence recognition by bispyrazinonaphthalimides antitumor agents. AB - Bifunctional DNA intercalating agents have long attracted considerable attention as anticancer agents. One of the lead compounds in this category is the dimeric antitumor drug elinafide, composed of two tricyclic naphthalimide chromophores separated by an aminoalkyl linker chain optimally designed to permit bisintercalation of the drug into DNA. In an effort to optimize the DNA recognition capacity, different series of elinafide analogues have been prepared by extending the surface of the planar drug chromophore which is important for DNA sequence recognition. We report here a detailed investigation of the DNA sequence preference of three tetracyclic monomeric or dimeric pyrazinonaphthalimide derivatives. Melting temperature measurements and surface plasmon resonance (SPR) studies indicate that the dimerization of the tetracyclic planar chromophore considerably augments the affinity of the drug for DNA, polynucleotides, or hairpin oligonucleotides and promotes selective interaction with G.C sites. The (CH(2))(2)NH(CH(2))(3)NH(CH(2))(2) connector stabilizes the drug-DNA complexes. The methylation of the two nitrogen atoms of this linker chain reduces the binding affinity and increases the dissociation rates of the drug-DNA complexes by a factor of 10. DNase I footprinting experiments were used to investigate the sequence selectivity of the drugs, demonstrating highly preferential binding to G.C-rich sequences. It also served to select a high affinity site encompassing the sequence 5'-GACGGCCAG which was then introduced into a biotin-labeled hairpin oligonucleotide to accurately measure the binding parameters by SPR. The affinity constant of the unmethylated dimer for this sequence is 500 times higher than that of the monomer compound and approximately 10 times higher than that of the methylated dimer. The DNA groove accessibility was also probed with three related oligonucleotides carrying G --> c(7)G, G --> I, and C --> M substitutions. The level of drug binding to the two hairpin oligonucleotides containing 7-deazaguanine (c(7)G) or 5-methylcytosine (M) residues is unchanged or only slightly reduced compared to that of the unmodified target. In contrast, incorporation of inosine (I) residues considerably decreases the extent of drug binding or even abolishes the interaction as is the case with the monomer. The pyrazinonaphthalimide derivatives are thus much more sensitive to the deletion of the exocyclic guanine 2-amino group exposed in the minor groove of the duplex than to the modification of the major groove elements. The complementary SPR footprinting methodology combining site selection and quantitative DNA affinity analysis constitutes a reliable method for dissecting the DNA sequence selectivity profile of reversible DNA binding small molecules. PMID- 14529287 TI - Fatty acid-binding proteins inhibit hydration of epoxyeicosatrienoic acids by soluble epoxide hydrolase. AB - Epoxyeicosatrienoic acids (EETs) are potent regulators of vascular homeostasis and are bound by cytosolic fatty acid-binding proteins (FABPs) with K(d) values of approximately 0.4 microM. To determine whether FABP binding modulates EET metabolism, we examined the effect of FABPs on the soluble epoxide hydrolase (sEH)-mediated conversion of EETs to dihydroxyeicosatrienoic acids (DHETs). Kinetic analysis of sEH conversion of racemic [(3)H]11,12-EET yielded K(m) = 0.45 +/- 0.08 microM and V(max) = 9.2 +/- 1.4 micromol min(-1) mg(-)(1). Rat heart FABP (H-FABP) and rat liver FABP were potent inhibitors of 11,12-EET and 14,15 EET conversion to DHET. The resultant inhibition curves were best described by a substrate depletion model, with K(d) = 0.17 +/- 0.01 microM for H-FABP binding to 11,12-EET, suggesting that FABP acts by reducing EET availability to sEH. The EET depletion by FABP was antagonized by the co-addition of arachidonic acid, oleic acid, linoleic acid, or 20-hydroxyeicosatetraenoic acid, presumably due to competitive displacement of FABP-bound EET. Collectively, these findings imply that FABP might potentiate the actions of EETs by limiting their conversion to DHET. However, the effectiveness of this process may depend on metabolic conditions that regulate the levels of competing FABP ligands. PMID- 14529288 TI - Lipid-induced conformational switch in the membrane binding domain of CTP:phosphocholine cytidylyltransferase: a circular dichroism study. AB - CTP:phosphocholine cytidylyltranferase (CCT) regulates phosphatidylcholine (PC) biosynthesis. Its activity is controlled by reversible interactions with membrane lipids, mediated by an internal segment referred to as domain M. Although domain M peptides adopt an amphipathic alpha-helical structure when membrane bound, the structure of this domain in the context of the whole enzyme in the lipid-free and lipid-bound state is unknown. Here we derive lipid-induced secondary structural changes in CCTalpha using circular dichroism and three deconvolution programs. The analysis of two fragments, CCT236 (CCT1-236, housing the catalytic domain) and a synthetic domain M peptide (CCT237-293) aided the assignment of structural change to specific domains. To carry out this study, we developed a micellar lipid activating system that would avoid generation of CCT-induced lipid vesicle aggregates that interfere with the CD analysis. Lysophosphatidylcholine/phosphatidylglycerol (LPC/PG) mixed micelles supported full activation of CCT and caused an increase in the alpha-helix content of full length CCT from 25 to 41%, at the expense of all other conformations. LPC/PG also induced an increase in alpha-helix content of the domain M peptide from 7 to 85% at the expense of all other conformers. This lipid system did not significantly affect the secondary structure of CCT236, nor did it affect the proteolytic fragmentation pattern of this region within full-length CCT, suggesting that the region containing the catalytic domain changes very little upon membrane activation of CCT. Our data suggest that lipids trigger a conformational switch in domain M from a mixed structure to an alpha-helix, thus creating a hydrophobic face for membrane insertion. Our results negate the idea that domain M is entirely helical in both the soluble and membrane-bound forms of CCT. PMID- 14529289 TI - Flexible metal binding of the metallo-beta-lactamase domain: glyoxalase II incorporates iron, manganese, and zinc in vivo. AB - Glyoxalase II belongs to the metallo-beta-lactamase superfamily of proteins, possessing the characteristic dinuclear active site. Within this protein family, glyoxalase II from Arabidopsis thaliana is the first member to be isolated with significant amounts of iron, manganese, and zinc when being recombinantly produced in Escherichia coli. Enzyme preparations with different ratios of these three metals all yield k(cat)/K(M) values in the range of 1.5-1.9 s(-1) microM( 1) with the substrate S-d-lactoylglutathione. X-ray absorption spectroscopy reveals binding of all three metals to the dinuclear active site with 5-6-fold coordination consisting of 2.5 +/- 0.5 histidine and 2.5 +/- 0.5 oxygen ligands. This model does not distinguish site-specific or distributed binding. The metal metal distance is determined to be 3.18 +/- 0.06 A. Electron paramagnetic resonance spectroscopy gives evidence for several different types of dimetal sites, including spin-coupled Fe(III)Fe(II), Fe(III)Zn(II), and Mn(II)Mn(II) centers. The metal-ligand distances measured by X-ray absorption spectroscopy vary depending on the metal type and comply with their element-specific, characteristic values. This reflects a high degree of structural flexibility within the glyoxalase II dinuclear active site, which is considered as the structural basis for its broad metal selectivity. PMID- 14529290 TI - Aromatic thiol pKa effects on the folding rate of a disulfide containing protein. AB - The production of proteins via recombinant DNA technology often requires the in vitro folding of inclusion bodies, which are protein aggregates. To create a more efficient redox buffer for the in vitro folding of disulfide containing proteins, aromatic thiols were investigated for their ability to increase the folding rate of scrambled RNase A. Scrambled RNase A is fully oxidized RNase A with a relatively random distribution of disulfide bonds. The importance of the thiol pK(a) value was investigated by the analysis of five para-substituted aromatic thiols with pK(a) values ranging from 5.2 to 6.6. Folding was measured at pH 6.0 where the pK(a) value of the thiols would be higher, lower, or equal to the solution pH. Thus, relative concentrations of thiol and thiolate would vary across the series. At pH 6.0, the aromatic thiols increased the folding rate of RNase A by a factor of 10-23 over that observed for glutathione, the standard additive. Under optimal conditions, the apparent rate constant increased as the thiol pK(a) value decreased. Optimal conditions occurred when the concentration of protonated thiol in solution was approximately 2 mM, although the total thiol concentration varied considerably. The importance of the concentration of protonated thiol in solution can be understood based on equilibrium effects. Kinetic studies suggest that the redox buffer participates as the nucleophile and/or the center thiol in the key rate determining thiol disulfide interchange reactions that occur during protein folding. Aromatic thiols proved to be kinetically faster and more versatile than classical aliphatic thiol redox buffers. PMID- 14529291 TI - Purification of recombinant human cPLA2 gamma and identification of C-terminal farnesylation, proteolytic processing, and carboxymethylation by MALDI-TOF-TOF analysis. AB - Cytosolic phospholipase A(2)gamma (cPLA(2)gamma) is a calcium-independent, membrane-associated phospholipase A(2) that possesses a C-terminal prenylation motif (-CCLA) whose covalent structure cannot be deduced from the primary sequence alone. Accordingly, we overexpressed human cPLA(2)gamma containing an N terminal His tag ((His)(6)cPLA(2)gamma) in Sf9 cells and quantitatively solubilized and purified the enzyme by sequential immobilized metal affinity and Mono Q column chromatographies. The final preparation appeared as a single 61 kDa band after SDS-PAGE/silver-staining, possessed high lysophospholipase activity (50 micromol min(-1) mg(-1)), and was inhibited by, but did not hydrolyze, palmitoyl-CoA. Radiolabeling of recombinant human cPLA(2)gamma with [(3)H] mevalonolactone in the absence of statins and subsequent cleavage of prenyl groups with Raney nickel revealed that the enzyme is only farnesylated and is not geranylgeranylated. Analysis of CNBr-digested cPLA(2)gamma by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI/TOF-TOF) mass spectrometry demonstrated the presence of a farnesyl moiety at Cys-538, cleavage of the Cys(538)-Cys(539) bond, and carboxymethylation of the resultant C-terminal prenylated cysteine. Collectively, these results describe the solubilization and purification of recombinant cPLA(2)gamma to homogeneity and identify cPLA(2)gamma as a farnesylated protein that undergoes at least three sequential posttranslational modifications that likely facilitate its targeting and interactions with its membrane substrates. PMID- 14529292 TI - Cholesterol depletion inhibits store-operated calcium currents and exocytotic membrane fusion in RBL-2H3 cells. AB - The effects of cholesterol depletion from the plasma membrane with methyl-beta cyclodextrin (MbetaCD) on exocytotic processes were investigated in rat basophil leukemia cells (RBL-2H3 cells). Pretreatment of the cells with MbetaCD inhibited antigen-evoked exocytotic release dose-dependently. To elucidate the mechanism of this inhibition, we performed experiments on the effects of MbetaCD on exocytotic membrane fusion and mobilization of Ca(2+) and on the localization of the tyrosine kinase Lyn. Inhibition of degranulation by MbetaCD was observed even under stimulation with the phorbol ester and calcium ionophore. Therefore, MbetaCD affected a process downstream of Ca(2+) influx, or membrane fusion between the granule and the plasma membrane. Intracellular calcium measurements revealed that MbetaCD inhibited the Ca(2+) increase induced by antigen. Furthermore, we found that MbetaCD significantly inhibited Ca(2+) influx from the extracellular medium through the store-operated calcium channel (SOC) but did not affect Ca(2+) release from the intracellular Ca(2+) store. Fluorescent image analysis of cells expressing Lyn-YFP showed that treatment with MbetaCD scarcely affected the localization and lateral mobility of Lyn in the plasma membrane. These results suggest that cholesterol depletion by MbetaCD decreases degranulation mainly by inhibiting the SOC and membrane fusion between the secretory granules and the plasma membrane in mast cells. PMID- 14529294 TI - RPE65 operates in the vertebrate visual cycle by stereospecifically binding all trans-retinyl esters. AB - RPE65 is a major protein of unknown function found associated with the retinyl pigment epithelial (RPE) membranes [Hamel, C. P., Tsilou, E., Pfeffer, B. A., Hooks, J. J., Detrick, B., and Redmond, T. M. (1993) J. Biol. Chem. 268, 15751 15757; Bavik, C. O., Levy, F., Hellman, U., Wernstedt, C., and Eriksson, U. (1993) J. Biol. Chem. 268, 20540-20546]. RPE65 knockouts fail to synthesize 11 cis-retinal, the chromophore of rhodopsin, and accumulate all-trans-retinyl esters in the RPE. Previous studies have also shown that RPE65 is specifically labeled with all-trans-retinyl ester based affinity labeling agents, suggesting a retinyl ester binding role for the protein. In the present work, we show that purified RPE65 binds all-trans-retinyl palmitate (tRP) with a K(D) = 20 pM. These quantitative experiments are performed by measuring the quenching of RPE65 fluorescence by added tRP. The binding for tRP is highly specific because 11-cis retinyl palmitate binds with a K(D) = 14 nM, 11-cis-retinol binds with a K(D) = 3.8 nM, and all-trans-retinol (vitamin A) binds with a K(D) = 10.8 nM. This stereospecificity for tRP is to be compared to the binding of retinoids to BSA, where virtually no discrimination is found in the binding of the same retinoids. This work provides further evidence that RPE65 functions by binding to and mobilizing the highly hydrophobic all-trans-retinyl esters, allowing them to enter the visual cycle. PMID- 14529293 TI - Creation of a fully functional cysteine-less variant of osmosensor and proton osmoprotectant symporter ProP from Escherichia coli and its application to assess the transporter's membrane orientation. AB - Transporter ProP of Escherichia coli is an osmosensor and an osmoprotectant transporter. Previous results suggest that medium osmolality determines the proportions of ProP in active and inactive conformations. A cysteine-less (Cys less) variant was created and characterized as a basis for structural and functional analyses based on site-directed Cys substitution and chemical labeling of ProP. Parameters describing the osmosensory and osmoprotectant transport activities of Cys-less ProP-(His)(6) variants were examined, including the threshold for osmotic activation and the absolute transporter activity at high osmolality (in both cells and proteoliposomes), the dependence of K(M) and V(max) for proline uptake on osmolality, and the rate constant for transporter activation in response to an osmotic upshift (in cells only). Variant ProP (His)(6)-C112A-C133A-C264V-C367A (designated ProP) retained similar activities to ProP-(His)(6) in both cells and proteoliposomes. The bulky Val residue was favored over Ala or Ser at position 264, whereas Val strongly impaired function when placed at position 367, highlighting the importance of residues at those positions for osmosensing. In the ProP* background, variants with a single Cys residue at positions 112, 133, 241, 264, 293, or 367 retained full function. The native Cys at positions 112, 133, 264, and 367, predicted to be within transmembrane segments of ProP, were poorly reactive with membrane-impermeant thiol reagents. The reactivities of Cys at positions 241 and 293 were consistent with exposure of those residues on the cytoplasmic and periplasmic surfaces of the cytoplasmic membrane, respectively. These observations are consistent with the topology and orientation of ProP predicted by hydropathy analysis. PMID- 14529295 TI - Calcium-dependent conformational change and thermal stability of the isolated PsbO protein detected by FTIR spectroscopy. AB - The structure and function of the photosystem II PsbO extrinsic protein is under intense research, being an essential part of the biomolecular engine that carries out water oxidation and oxygen production. This paper presents a structural analysis of the isolated PsbO protein by FTIR spectroscopy, reporting detailed secondary structure quantification and changes in the secondary structure content of the protein attributed to the effect of calcium (Ca(2+)). Measurements in H(2)O and D(2)O have allowed us to see the effect of calcium on the conformation of the protein. The results indicate that (i) the protein presents a major content of beta-structure (i.e., beta-sheet, beta-strands, beta-turns) as detected by the infrared bands at 1624-1625, 1678-1679, 1688-1689 cm(-1), which account for about 38% in water and 33% in heavy water, in the presence of calcium; and (ii) the amount of this beta-structure fraction increases 7-10% in the absence of calcium, with a concomitant decrease in loops and nonordered structure. The thermal denaturation profile of the protein in the presence of calcium showed low stability with T(m) approximately 56 degrees C. This profile also shows a second phase of denaturation above 60 degrees C and the appearance of aggregation signals above 70 degrees C. Our observations indicate that calcium is able to modify the conformation of the protein at least in solution and confirm that PsbO is mainly a beta-protein where beta-sheet is the major ordered secondary structure element of the protein core. PMID- 14529296 TI - Control of energy dissipation and photochemical activity in photosystem I by NADP dependent reversible conformational changes. AB - Addition of NADP(+) to thylakoid membranes or isolated photosystem I (PSI) submembrane fractions quenched chlorophyll fluorescence by up to 40% at low or room temperature. This quenching was reversed by NADPH. Similar quenching was also observed with the addition of heparin or thenoyltrifluoroacetone (TTFA), inhibitors that bind ferredoxin:NADP(+) reductase (FNR) and prevent reduction of NADP(+). The NADP(+)-induced quenching coincided with a reversible conformational change of the secondary protein structure in the PSI submembrane fractions where 20% of the alpha-helix conformations were transformed mainly into beta-sheet-like structures. Further, P700 photooxidation was retarded due to this conformational change, and about 25% of the centers could not be photooxidized, these changes being also reversible with addition of NADPH. The above modifications in the presence of NADP(+) also increased photodamage processes under strong illumination, and NADPH protected it. Conformational modification of FNR upon binding of NADP(+) or NADPH is proposed to trigger the macromolecular changes in a larger part of the protein complex of PSI. The conformational changes must increase the intermolecular distances and change the mutual orientation between the various cofactors in the PSI complex. This new control mechanism of energy dissipation and photochemical activity by NADP(+)/NADPH is proposed to increase the turnover rate of PSI under conditions when both linear and cyclic electron transport activities must be supported. PMID- 14529297 TI - Probing hydrogen-bonding interactions in the active site of medium-chain acyl-CoA dehydrogenase using Raman spectroscopy. AB - The role of the oxyanion hole in the reaction catalyzed by pig medium-chain acyl CoA dehydrogenase (pMCAD) has been investigated using enzyme reconstituted with 2'-deoxy-FAD. The k(cat) (18.8 +/- 0.5 s(-1)) and K(m) (2.5 +/- 0.4 microM) values for the oxidation of n-octanoyl-CoA (C(8)-CoA) by WT pMCAD recombinantly expressed in Escherichia coli are similar to those of native pMCAD isolated from pig kidney. In agreement with previous studies [Engst et al. (1999) Biochemistry 38, 257-267], reconstitution of the WT enzyme with 2'-deoxy-FAD causes a large (400-fold) decrease in k(cat) but has little effect on K(m). To investigate the molecular basis for the alterations in activity resulting from changes in hydrogen bonding between the substrate and the enzyme's oxyanion hole, the structure of the product analogue hexadienoyl-CoA (HD-CoA) bound to the 2'-deoxy FAD-reconstituted enzyme has been probed by Raman spectroscopy. Importantly, while WT pMCAD causes a 27 cm(-1) decrease in the vibrational frequency of the HD enone band, from 1595 to 1568 cm(-1), the enone band is only shifted 10 cm(-1) upon binding HD-CoA to 2'-deoxy-FAD pMCAD. Thus, removal of the 2'-ribityl hydroxyl group results in a substantial reduction in the ability of the enzyme to polarize the ground state of the ES complex. On the basis of an analysis of a similar system, it is estimated that ground state destabilization is reduced by up to 17 kJ mol(-1), while the activation energy for the reaction is raised 15 kJ mol(-1). In addition, removal of the 2'-ribityl hydroxyl reduces the redox potential shift that is induced by HD-CoA binding from 18 to 11 kJ mol(-1). Consequently, while ligand polarization caused by hydrogen bonding in the oxyanion hole is intimately linked to substrate turnover, additional factors must be responsible for ligand-induced changes in redox potential. Finally, while replacement of the catalytic base E376 with Gln abolishes the ability of the enzyme to catalyze substrate oxidation and to catalyze the exchange of the C(8) CoA alpha-protons with solvent deuterium, the 2'-deoxy-FAD-reconstituted enzyme catalyzes alpha-proton exchange at a rate (k(exc)) of 0.085 s(-1), which is only 4-fold slower than k(exc) for WT pMCAD (0.35 s(-1)). Thus, either the oxyanion hole plays only a minor role in stabilizing the transition state for alpha-proton exchange, in contrast to its role in substrate oxidation, or the value of k(exc) for WT pMCAD reflects a process such as exchange of the E376 COOH proton with solvent. PMID- 14529298 TI - Influence of the C-terminal residues on oligomerization of alpha A-crystallin. AB - Earlier studies have shown that the chaperone activity of alpha-crystallin is significantly affected in diabetic rat and human lenses. Subsequently, mass spectrometric analysis showed diabetic lenses having high levels of the alphaA crystallins in which different numbers of C-terminal residues were deleted. The present study was aimed to show whether cleavage of these residues influences protein structure, oligomerization, and chaperone function. For generation of various mutants, a stop codon was introduced at the positions of interest, proteins were expressed in BL21(DE3)pLys S E. coli, and the truncated alphaA crystallins were purified by size-exclusion chromatography. The molecular masses, as determined by molecular sieve HPLC, of mutants with deletions of 1, 5, and 10 C-terminal residues (group-1) were 519-602 kDa, and those of mutants with deletions of 11, 16, and 22 C-terminal residues (group-2) were 148-152 kDa, as compared to 607 kDa for alphaA-wild type. On the basis of circular dichroism measurements, the alpha helix content was 2-fold higher and the tertiary structure was significantly altered in the group-2 mutants. Chaperoning abilities, as determined by the ADH assay and the betaL-crystallin heat denaturation assay, of the group-1 mutants, with the exception of alphaA(1-163), were slightly improved or unchanged, that of alphaA(1-163) was moderately affected, and those of the group-2 mutants were severely affected. Most strikingly, cleavage of 11 C-terminal residues including Arg-163 showed a substantial decrease in oligomeric size and chaperone function and significant changes in protein structure whereas cleavage of 10 residues had either a small effect or no effect at all. This points to an important role for the C-terminal extension, Arg-163 in particular, and no significant role for the C-terminal flexible tail in the oligomer assembly of alphaA-crystallin. PMID- 14529299 TI - Tertiary conformation of the template-primer and gapped DNA substrates in complexes with rat polymerase beta. Fluorescence energy transfer studies using the multiple donor-acceptor approach. AB - The tertiary structure of template-primer and gapped DNA substrates in the complex with rat polymerase beta (pol beta) has been examined using the fluorescence energy transfer method based on the multiple donor-acceptor approach. In these studies, we used DNA substrates labeled at the 5' end of the template strand and the 5' end of the primer with the fluorescent donor and/or acceptor. Measurements of the enzyme complex with the template-primer DNA substrate having a ten nucleotide long ssDNA extension indicate that the distance between the 5' end of the template strand and the 5' end of the primer decreases by approximately 9.8 A as compared to the free nucleic acid. Analogous experiments with the template-primer substrate, having the ssDNA extension with five nucleotide residues, show approximately 6.6 A distance decrease. Such large distance decreases indicate that the DNA is significantly bent in the binding site. Analysis of the data indicates that the bending occurs between the third and the fourth nucleotide of the ssDNA extension. The entire template strand is at the bend angle Theta(TP) = 85 +/- 7 degrees with respect to the dsDNA part of the DNA molecule. In the polymerase complex with the gapped DNA, the distance between the 5' ends of the DNA and the bend angle are 66 +/- 2.2 A and 65 +/- 6 degrees, respectively. These values are very similar to the same distance and bend angle of the gap complex in the crystal structure of the co-complex. The presence of the 5'-terminal PO(4)(-) group downstream from the primer does not affect the tertiary conformation of the gapped DNA, indicating that the effect of the phosphate group is localized at the ssDNA gap. PMID- 14529300 TI - Solid-state nuclear magnetic resonance evidence for parallel and antiparallel strand arrangements in the membrane-associated HIV-1 fusion peptide. AB - The HIV-1 fusion peptide serves as a useful model system for understanding viral/target cell fusion, at least to the lipid-mixing stage. Previous solid state NMR studies have shown that the membrane-bound HIV-1 fusion peptide adopts an extended conformation in a lipid mixture close to that of host cells of the virus. In the present study, solid-state NMR REDOR methods were applied for detection of oligomeric beta strand structure. The samples were prepared under fusogenic conditions and contained equimolar amounts of two peptides, one with selective [(13)C]carbonyl labeling and the other with selective [(15)N]amide labeling. In the REDOR measurements, observation of reduced (13)C intensity due to hydrogen-bonded amide (15)N provides strong experimental evidence of oligomer formation by the membrane-associated peptide. Comparison of REDOR spectra on samples that were labeled at different residue positions suggests that there are both parallel and antiparallel arrangements of peptide strands. In the parallel arrangement, interpeptide hydrogen bonding decreases toward the C-terminus, while in the antiparallel arrangement, hydrogen bonds are observed along the entire length of residues which was probed (Gly-5 to Gly-16). For the parallel arrangement, these observations are consistent with the model in which the apolar N-terminal and central regions of the peptides penetrate into the membrane and hydrogen bond with one another while the polar C-terminus of the peptide is outside the membrane and hydrogen bonds with water. These measurements show that, at least at the end state of fusion, the peptide can adopt an oligomeric beta strand structure. PMID- 14529301 TI - Foreword: Hadron therapy--from yesterday's physics laboratory to today's modern clinical routine. PMID- 14529302 TI - Boron neutron capture therapy: cellular targeting of high linear energy transfer radiation. AB - Boron neutron capture therapy (BNCT) is based on the preferential targeting of tumor cells with (10)B and subsequent activation with thermal neutrons to produce a highly localized radiation. In theory, it is possible to selectively irradiate a tumor and the associated infiltrating tumor cells with large single doses of high-LET radiation while sparing the adjacent normal tissues. The mixture of high and low-LET dose components created in tissue during neutron irradiation complicates the radiobiology of BNCT. Much of the complexity has been unravelled through a combination of preclinical experimentation and clinical dose escalation experience. Over 350 patients have been treated in a number of different facilities worldwide. The accumulated clinical experience has demonstrated that BNCT can be delivered safely but is still defining the limits of normal brain tolerance. Several independent BNCT clinical protocols have demonstrated that BNCT can produce median survivals in patients with glioblastoma that appear to be equivalent to conventional photon therapy. This review describes the individual components and methodologies required for effect BNCT: the boron delivery agents; the analytical techniques; the neutron beams; the dosimetry and radiation biology measurements; and how these components have been integrated into a series of clinical studies. The single greatest weakness of BNCT at the present time is non uniform delivery of boron into all tumor cells. Future improvements in BNCT effectiveness will come from improved boron delivery agents, improved boron administration protocols, or through combination of BNCT with other modalities. PMID- 14529303 TI - Heavy ion therapy: status and perspectives. AB - Starting with the pioneering work at the University of California in Berkeley in 1977, heavy ion radiotherapy has been of increasing interest especially in Japan and Europe in the last decade. There are currently 3 facilities treating patients with carbon ions, two of them in Japan within a clinical setting. In Germany, a research therapy facility is in operation and the construction of a new hospital based facility at the Heidelberg university will be started soon. An outline of the current status of heavy ion radiotherapy is given with emphasis to the technical aspects of the respective facilities. This includes a description of passive and active beam shaping systems, as well as their implications for treatment planning and dosimetry. The clinical trials and routine treatments performed at the German heavy ion facility are summarized. An overview over the upcoming new facilities and their technical possibilities is given. It is discussed what the necessary improvements are to fully exploit the potential of these facilities. Especially the new Heidelberg facility with the possibility of active beam scanning in combination with the first isocentric gantry for ions and offering beams of protons, helium, oxygen and carbon ions has implications on treatment planning, dosimetry and quality assurance. The necessary and ongoing developments in these areas are summarized. The new facilities also offer the possibilities to perform more extensive clinical studies and to explore future indications for radiotherapy with heavy ions. An overview over the indications and treatment schemes is also given. PMID- 14529304 TI - Treatment planning for conformal proton radiation therapy. AB - Clinical results from various trials have demonstrated the viability of protons in radiation therapy and radiosurgery. This has motivated a few large medical centers to design and build expensive hospital based proton facilities based proton facilities (current cost estimates for a proton facility is around 100 million US dollars). Until this development proton therapy was done using retrofitted equipment originally designed for nuclear experiments. There are presently only three active proton therapy centers in the United States, 22 worldwide. However, more centers are under construction and being proposed in the US and abroad. The important difference between proton and x-ray therapy is in the dose distribution. X-rays deposit most of their dose at shallow depths of a few centimeters with a gradual decay with depth in the patient. Protons deliver most of their dose in the Bragg peak, which can be delivered at most clinically required depths followed by a sharp fall-off. This sharp falloff makes protons sensitive to variations in treatment depths within patients. Treatment planning incorporates all the knowledge of protons into a process, which allows patients to be treated accurately and reliably. This process includes patient immobilization, imaging, targeting, and modeling of planned dose distributions. Although the principles are similar to x-ray therapy some significant differences exist in the planning process, which described in this paper. Target dose conformality has recently taken on much momentum with the advent of intensity modulated radiation therapy (IMRT) with photon beams. Proton treatments provide a viable alternative to IMRT because they are inherently conformal avoiding normal tissue while irradiating the intended targets. Proton therapy will soon bring conformality to a new high with the development of intensity modulated proton therapy (IMPT). Future challenges include keeping the cost down, increasing access to conventional proton therapy as well as the clinical implementation of IMPT. Computing advances are making Monte Carlo techniques more accessible to treatment planning for all modalities including proton therapy. This technique will allow complex delivery configurations to be properly modeled in a clinical setting. PMID- 14529305 TI - Optimization of physical dose distributions with hadron beams: comparing photon IMRT with IMPT. AB - Intensity modulated radiotherapy with high enery photons (IMRT) and with charged particles (IMPT) refer to the most advanced development in conformal radiation therapy. Their general aim is to increase local tumor control rates while keeping the radiation induced complications below desired thresholds. IMRT is currently widely introduced in clinical practice. However, the more complicated IMPT is still under development. Especially, spot- scanning techniques integrated in rotating gantries that can deliver proton or light ion-beams to a radiation target from any direction will be available in the near future. We describe the basic concepts of intensity modulated particle therapy (IMPT). Starting from the potential advantages of hadron therapy inverse treatment planning strategies are discussed for various dose delivery techniques of IMPT. Of special interest are the techniques of distal edge tracking (DET) and 3D-scanning. After the introduction of these concepts a study of comparative inverse treatment planning is presented. The study aims to identify the potential advantages of achievable physical dose distributions with proton and carbon beams, if different dose delivery techniques are employed. Moreover, a comparison to standard photon IMRT is performed. The results of the study are summarized as: i) IMRT with photon beams is a strong competitor to intensity modulated radiotherapy with charged particles. The most obvious benefit observed for charged particles is the reduction of medium and low doses in organs at risk. ii) The 3D-scanning technique could not improve the dosimetric results achieved with DET, although 10 15 times more beam spots were employed for 3D-scanning than for DET. However, concerns may arise about the application of DET, if positioning errors of the patient or organ movements have to be accounted for. iii) Replacing protons with carbon ions leads to further improvements of the physical dose distributions. However, the additional degree of improvement due to carbon ions is modest. The main clinical potential of heavy ion beams is probably related to their radiobiological properties. PMID- 14529306 TI - Significance and implementation of RBE variations in proton beam therapy. AB - Key to radiation therapy is to apply a high tumor-destroying dose while protecting healthy tissue, especially near organs at risk. To optimize treatment for ion therapy not the dose but the dose multiplied by the relative biological effectiveness (RBE) is decisive. Proton therapy has been based on the use of a generic RBE, which is applied to all treatments independent of dose/fraction, position in the spread-out Bragg peak (SOBP), initial beam energy or the particular tissue. Dependencies of the RBE on various physical and biological properties are disregarded. The variability of RBE in clinical situations is believed to be within 10-20%. This is in the same range of effects that receive high attention these days, i.e., patient set-up uncertainties, organ motion effects, and dose calculation accuracy all affecting proton as well as conventional radiation therapy. Elevated RBE values can be expected near the edges of the target, thus probably near critical structures. This is because the edges show lower doses and, depending on the treatment plan, may be identical with the beam's distal edge, where dose is deposited in part by high-LET protons. We assess the rationale for the continued use of a generic RBE and whether the magnitude of RBE variation with treatment parameters is small relative to our abilities to determine RBE's. Two aspects have to be considered. Firstly, the available information from experimental studies and secondly, our ability to calculate RBE values for a given treatment plan based on parameters extracted from such experiments. We analyzed published RBE values for in vitro and in vivo endpoints. The values for cell survival in vitro indicate a substantial spread between the diverse cell lines. The average value at mid SOBP over all dose levels is approximately 1.2 in vitro and approximately 1.1 in vivo. Both in vitro and in vivo data indicate a statistically significant increase in RBE for lower doses per fraction, which is much smaller for in vivo systems. The experimental in vivo data indicate that continued employment of a generic RBE value of 1.1 is reasonable. At present, there seems to be too much uncertainty in the RBE value for any human tissue to propose RBE values specific for tissue, dose/fraction, etc. There is a clear need for prospective assessments of normal tissue reactions in proton irradiated patients and determinations of RBE values for several late responding tissues in animal systems, especially as a function of dose in the range of 1-4 Gy. However, there is a measurable increase in RBE over the terminal few mm of the SOBP, which results in an extension of the bio-effective range of the beam of a few mm. This needs to be considered in treatment planning, particularly for single field plans or for an end of range in or close to a critical structure. To assess our ability to calculate RBE values we studied two approaches, which are both based on the track structure theory of radiation action. RBE calculations are difficult since both the physical input parameters, i.e., LET distributions, and, even more so, the biological input parameters, i.e., local cellular response, have to be known with high accuracy. Track structure theory provides a basis for predicting dose-response curves for particle irradiation. However, designed for heavy ion applications the models show weaknesses in the prediction of proton radiation effects. We conclude that, at present, RBE modeling in treatment planning involves significant uncertainties. To incorporate RBE variations in treatment planning there has to be a reliable biological model to calculate RBE values based on the physical characteristics of the radiation field and based on well-known biological input parameters. In order to do detailed model calculations more experimental data, in particular for in vivo endpoints, are needed PMID- 14529307 TI - The increased biological effectiveness of heavy charged particles: from radiobiology to treatment planning. AB - The increased biological effectiveness of heavy charged particle beams like e.g., carbon ions in the tumor volume in comparison to the lower effectiveness in the surrounding healthy tissue represents one of the major rationales for their application in tumor therapy. This increased effectiveness also characterizes the advantage of heavier ions compared to proton beams. The increased effectiveness has to be taken into account in treatment planning in order to estimate the corresponding photon equivalent doses in normal and tumor tissues, thus allowing a link e.g., to normal tissue dose limits in conventional photon therapy. Due to the complex dependencies of RBE on parameters like dose, beam energy, LET, atomic number and cell or tissue type, the relevant RBEs cannot be solely determined from experimental data. Therefore, within the framework of the pilot project of tumor therapy with carbon ions performed at GSI Darmstadt, treatment planning is based on a biophysical model, which has been extensively tested. The paper first summarizes the essential systematic dependencies of RBE on different parameters like e.g., dose, LET, atomic number and cell type. The basic principle of the biophysical model is then introduced, and special emphasis is given to the application of the model to in vivo and clinical endpoints. Model predictions are compared to experimental data in vitro and in vivo. Finally, the implementation of the biophysical model in the treatment planning procedure is presented. The biological verification of the whole treatment planning procedure is explained and examples of patient treatment plans are given. PMID- 14529308 TI - Optimization of beam parameters and treatment planning for intensity modulated proton therapy. AB - One of the objectives of the ongoing research and development work at the Northeast Proton Therapy Center (NPTC) in Boston is to perform optimized intensity modulated proton therapy (IMPT) treatments. Such treatments may be delivered by magnetically scanning a narrow proton pencil beam across the target volume, while both the scanning speed and the intensity of the beam are modulated. Localization of the proton dose in space allows one to yield dose distributions that are highly conformal to the target volume, thus minimizing the dose delivered to the surrounding healthy tissue. The aim of the current research is to determine technically optimal and clinically relevant specifications for the scanned beam delivery system, which is being developed in collaboration with Ion Beam Applications (IBA); and to create a link between the treatment planning and the beam delivery. IMPT treatment planning is performed for patient cases treated at the NPTC, with KonRad Pro software developed at the German Cancer Research Center (DKFZ). For the IMPT delivery, the proton intensity maps, optimized for discrete pencil beam spots, need to be translated into continuous scanning patterns. At the same time it is necessary to minimize the discrepancy between the planned and delivered doses which may result from such conversion, as well as from the technical limitations of the delivery system. Possibilities have been investigated for improving the proton dose conformity by optimizing the beam and scanning nozzle parameters, and by taking the specifics and limitations of the system into account in the treatment planning stage. PMID- 14529309 TI - Range, range modulation, and field radius requirements for proton therapy of prostate cancer. AB - The Loma Linda University Proton Treatment Facility has treated over 5,000 patients for prostate cancer. Other institutions may find information regarding field size and range requirements for this population of patients useful for designing new proton beamlines. The maximum range, range modulation, and maximum field radius for 240 fields of prostate patients undergoing treatment were sampled and analyzed. Most fields required a range less than 290 mm of water, a modulation width less than or equal to 120 mm, and a radius less than 75 mm. PMID- 14529310 TI - Use of the Leksell Gamma Knife for localized small field lens irradiation in rodents. AB - For most basic radiobiological research applications involving irradiation of small animals, it is difficult to achieve the same high precision dose distribution realized with human radiotherapy. The precision for irradiations performed with standard radiotherapy equipment is +/-2 mm in each dimension, and is adequate for most human treatment applications. For small animals such as rodents, whose organs and tissue structures may be an order of magnitude smaller than those of humans, the corresponding precision required is closer to +/-0.2 mm, if comparisons or extrapolations are to be made to human data. The Leksell Gamma Knife is a high precision radiosurgery irradiator, with precision in each dimension not exceeding 0.5 mm, and overall precision of 0.7 mm. It has recently been utilized to treat ocular melanoma and induce targeted lesions in the brains of small animals. This paper describes the dosimetry and a technique for performing irradiation of a single rat eye and lens with the Gamma Knife while allowing the contralateral eye and lens of the same rat to serve as the "control". The dosimetry was performed with a phantom in vitro utilizing a pinpoint ion chamber and thermoluminescent dosimeters, and verified by Monte Carlo simulations. We found that the contralateral eye received less than 5% of the administered dose for a 15 Gy exposure to the targeted eye. In addition, after 15 Gy irradiation 15 out of 16 animals developed cataracts in the irradiated target eyes, while 0 out of 16 contralateral eyes developed cataracts over a 6-month period of observation. Experiments at 5 and 10 Gy also confirmed the lack of cataractogenesis in the contralateral eye. Our results validate the use of the Gamma Knife for cataract studies in rodents, and confirmed the precision and utility of the instrument as a small animal irradiator for translational radiobiology experiments. PMID- 14529311 TI - 3D-ultrasound guided radiation therapy in the post-prostatectomy setting. AB - Daily transabdominal ultrasound-directed localization has proven valuable in correcting for setup error and organ motion in the treatment of prostate cancer with three-dimensional conformal radiation therapy (3DCRT). The present study sought to determine whether this transabdominal ultrasound technology could also be reliably applied in the post-operative adjuvant or salvage setting to improve the reproducibility of coverage of the intended volumes and to enhance conformal avoidance of adjacent normal structures. Sixteen consecutive patients who received external beam radiotherapy underwent daily localization using an optically guided 3D-ultrasound target localization system (SonArray, Zmed, Inc., Ashland, MA). Six of the above patients were treated in a post-prostatectomy setting, either adjuvantly or for salvage, while the remaining 10 with intact prostates were treated definitively. Because the bladder neck generally approximates the postoperative prostatic fossa, it was used during ultrasound localization as the primary reference structure for the post-prostatectomy patients. For patients treated definitively, the prostate was the primary reference structure. Daily shifts were recorded and port films were taken weekly immediately after ultrasound-based repositioning. By comparing port films taken after ultrasound localization, which evaluates for both set-up error and internal shift, with the original digitally reconstructed radiographs (DRRs), which represents a zero clinical set-up error situation, the degree of variability in organ position was determined. The average absolute, ultrasound-based shifts from the clinical set-up position in the anterior/posterior, lateral, and cranial/caudal directions for the post-prostatectomy patients were 5 +/- 4 mm SD, 3 +/- 3 mm SD, and 3 +/- 4 mm SD over the entire course of treatment, respectively. The average vector length shift was 8 +/- 4 mm SD. For patients treated with an intact prostate, the analogous average absolute shifts in the anterior/posterior, lateral, and cranial/caudal directions were 4 +/- 3 mm SD, 4 +/- 3 mm SD, and 4 +/- 3 mm SD over the entire course of treatment. The average vector length shift was 7 +/- 4 mm SD. Vector length shifts representing interfraction internal motion were estimated by comparing post-ultrasound port films with DRRs. These were 5 +/- 3 mm SD and 4 +/- 4mm SD for post-prostatectomy and intact prostate patients, respectively. These ultrasound-based displacements were not statistically different in patients with an intact prostate versus patients post-prostatectomy (p > 0.1). In conclusion, daily transabdominal 3D ultrasound localization proved to be a clinically feasible method of correcting for set-up and internal motion displacements. The bladder neck, which serves as an adequate localization reference structure for the prostatic fossa, could be readily ultrasound imaged and repositioned as necessary. Daily internal motion errors that would have occurred if only pre-treatment port films were used were similar in magnitude to those observed for the patients with intact prostates and were of sufficient magnitude to support the use of daily pre-treatment ultrasound localization in the post-prostatectomy setting. PMID- 14529312 TI - A model for evaluating therapeutic response of combined cancer treatment modalities: applied to treatment of subcutaneously implanted brain tumors (N32 and N29) in Fischer rats with pulsed electric fields (PEF) and 60Co-gamma radiation (RT). AB - The aim of the present study is to develop a mathematical model for evaluating therapeutic response of combined treatment modalities. The study was performed in rats of the Fischer-344 strain with rat glioma N32 or N29 tumors implanted subcutaneously on the thigh of the hind leg. Pulsed electric fields, PEF, with 16 exponentially decaying pulses with a maximum electric field strength of 140 V/mm and t(1/e)= 1 ms were first applied to the tumors. Then within 5 min radiation therapy with (60)Co-gamma radiation, RT, was given in daily fractions of 5 Gy. The animals were arranged into one group of controls and 3 groups of different kind of treatments: PEF only, RT only or combination of PEF + RT. At about 4 weeks after inoculation, the tumors were given the treatment sessions during one week. In 2 experimental series with totally 52 rats with N32 tumors, of which 16 were controls, were given 4 sessions of PEF treatments and RT (totally 20 Gy). In a special experimental series with totally 56 rats with N32 tumors, of which 10 were controls, the different groups were given 1, 2, 3 or 4 treatment sessions respectively. Another strain of glioma tumor, N29 with 62 tumors of which 14 were controls was studied in 2 series given 4PEF + 4RT and 2PEF + 4RT respectively. Fitting the data obtained from consecutive measurements of tumor volume (TV) of each individual tumor to an exponential model TV = TV(0). exp[TGR.t] estimated the tumor growth rate (TGR % per day) after the first day of treatment (t = 0). The TGR of N32 tumors treated with the combination of 4PEF + 4RT are significantly decreased compared to the controls (p < 0.0001), compared to RT alone (p < 0.0001) and compared to PEF alone (p < 0.001). The combined treatment of N32 gives significant effect on the tumor growth rate after 2, 3 and 4 treatment session while RT alone seems to be most efficient after one treatment of 5 Gy and PEF alone is most efficient after 2 treatments at 2 consecutive days. The TGR of N29 tumors treated with the combination of 4PEF + 4RT are significantly decreased compared to the controls (p < 0.05) but the combination of 2PEF + 4RT was more effective (p < 0.005). The specific therapeutic effect STE is defined as the difference between the average tumor growth rates of controls and exposed tumors divided by the average tumor growth rate of the controls. With 4PEF treatments alone the average STE value was 0.32 for N32 tumors and 0 for N29; for 4RT alone the STE values were 0.29 and 0.42 respectively, and for combined treatments 4PEF + 4RT 0.67 and 0.17 respectively. For the N29 tumors treated with 2PEF + 4RT the STE value was 0.53. The therapeutic enhancement ratio, TER, value increase with the number of treatment sessions and the TER of the combined treatments is above 1 in two of the N32 series, which indicates a synergistic effect of 4PEF + 4RT. This work demonstrate the use of our model for analyzing the combination PEF + RT, but it can also be used for evaluation the therapeutic effects of combining RT with chemotherapy or immunogenetic therapy. PMID- 14529313 TI - Viral and non-viral vectors in gene therapy: technology development and clinical trials. AB - Gene therapy as part of modern molecular medicine holds great promise for the treatment of both acute and chronic diseases and has the potential to bring a revolutionary era to cancer treatment. Gene therapy has been named the medicine of the future. For the past 10 years various viral and non-viral vectors have been engineered for improved gene and drug delivery. Although various diseases have been targeted, cancer therapy has been addressed to a large extent because of the straight forward approach. Delivery of toxic or immunostimulatory genes by viral and non-viral vectors has been investigated and encouraging results have been obtained in animal models. A large number of clinical trials have been conducted with some highly promising outcome. We propose that combinations of viruses with liposomes or polymers will solve the problem of systemic viral delivery and tumor targeting, bringing a revolution in molecular medicine and in applications of gene therapy in humans. PMID- 14529314 TI - MECP2 mutations or polymorphisms in mentally retarded boys: diagnostic implications. AB - BACKGROUND: Among the well characterized X-linked conditions causing mental retardation, mutations in the methyl-CpG-binding protein 2 gene (MECP2) in Xq28 have been found in up to 85% of patients with Rett syndrome, a neurologic disorder which, in addition to other symptoms, severely affects higher cognitive functions in females. Mutations in the MECP2 gene are involved in a broad spectrum of phenotypes from classical Rett syndrome to mild intellectual difficulties in females and neonatal encephalopathy in males. Recently, mutations in the MECP2 gene were reported in males with non-specific mental retardation suggesting that defects in MECP2 could be responsible for up to 2% of X-linked mental retardation. METHODS: We screened by denaturing high-pressure liquid chromatography the entire coding region and flanking intronic sequences of the MECP2 gene in a cohort of 354 mentally retarded males found negative for an expansion across the FRAXA CGG repeat and in a family in which a boy and his sister were mentally retarded. RESULTS: We identified mainly silent polymorphisms within the MECP2 gene, together with four sequence alterations of unknown significance, i.e. three missense mutations (T197M, T228S, and P376S) and one substitution at position -19 in intron 3 (378-19delT). Further familial investigations allowed us to ruled out a pathogenic effect for the intronic variant, the T228S and the P376S missense mutations. CONCLUSIONS: These results confirm that MECP2 mutations in males are far more rare than initially thought and call for a careful evaluation of the pathogenicity of the MECP2 missense mutations identified in mentally retarded males before genetic counseling is proposed to the relatives. PMID- 14529315 TI - Isolation of pneumococcal DNA from nasopharyngeal samples for real-time, quantitative PCR: comparison of three methods. AB - BACKGROUND: Real-time PCR is a useful method for detecting and quantifying bacterial DNA in clinical samples. DNA extraction is a crucial step when performing quantitative PCR. METHODS: We compared three methods, QIAamp. The use of tradenames is for product identification purposes only and does not imply endorsement. DNA Mini kit, MagNAPure trade mark LC DNA Isolation Kit II together with PickPen trade mark magnetic particle transfer device, and KingFisher genomic DNA purification Kit with KingFisher mL instrument, for purification of Streptococcus pneumoniae DNA from 50 nasopharyngeal swab samples, collected into skim milk-tryptone-glucose-glycerin medium. Pneumococcal DNA was detected and quantified by real-time PCR and results were compared to culture findings. RESULTS: The 22 (44%) pneumococcal culture-positive specimens were all positive by PCR regardless of DNA extraction method used, except that one KingFisher extracted sample was positive only when repeatedly tested. Additionally, 71%, 57%, and 82% of the culture-negative samples were positive by real-time PCR when DNA was extracted by QIAamp, MagNAPure-PickPen, and KingFisher methods, respectively. The number of genome equivalents detected by real-time PCR varied, but was mainly low in culture-negative samples. The sensitivities of culture and real-time PCR were hence compared by analyzing different dilutions of a pneumococcal suspension. Real-time PCR detected significantly higher numbers of genome equivalents than the numbers of bacteria detected by culture. CONCLUSIONS: The results indicate that the DNA extraction method used for quantitative PCR should be evaluated and that real-time PCR is more sensitive than bacterial culture for detecting pneumococcus in nasopharyngeal swab samples. PMID- 14529316 TI - MxA gene expression analysis as an interferon-beta bioactivity measurement in patients with multiple sclerosis and the identification of antibody-mediated decreased bioactivity. AB - BACKGROUND: Interferon-beta (IFNbeta) has proven to be an important advance in the therapy of multiple sclerosis (MS), but optimal markers for bioactivity have not been identified. To accurately measure bioactivity in MS patients treated with IFNbeta, we developed and tested a real-time reverse transcriptase (RT)-PCR assay for gene expression of MxA, an IFNbeta-induced gene in the peripheral blood of patients treated with IFNbeta. METHODS: We compared IFNbeta-treated patients with MS to controls in expression of MxA relative to the house-keeping gene, GAPDH. 2'-5'oligoadenylate synthetase (OAS) gene expression was also tested by real-time RT-PCR on RNA from the same patient specimens. Anti-IFNbeta antibody was measured by ELISA and a cytopathic effect assay. RESULTS: Seven of 54 patients were found to have complete loss of bioactivity. MxA expression correlated well with OAS expression. All patients with lost bioactivity had high levels of binding antibodies or neutralizing antibodies. CONCLUSIONS: This is the first demonstration that a real-time RT-PCR assay can be used to monitor therapy with interferons. These data identify MxA mRNA as an excellent biomarker for INFbeta action on the IFN receptor, and clarify the relationship between anti IFNbeta antibodies and bioactivity in patients with MS treated with IFNbeta. PMID- 14529317 TI - Detection of ALK gene rearrangements in formalin-fixed, paraffin-embedded tissue using a fluorescence in situ hybridization (FISH) probe: a search for optimum conditions of tissue archiving and preparation for FISH. AB - BACKGROUND: It is widely known that the efficiency of fluorescence in situ hybridization (FISH) probes applied to formalin-fixed, paraffin-embedded tissues is affected by the conditions under which the tissues are fixed and embedded. However, relatively few studies address exactly how tissue archiving conditions affect the performance of FISH probes. We report our experience based on use of an ALK FISH probe, during the validation of its diagnostic utility. METHODS: We applied the probe to 77 formalin-fixed, paraffin-embedded tissue blocks archived from 1991 through to 2000, and studied the interrelationship between the archival age (which ranged up to 10 years), type and condition of tissue, duration required for optimum hydrolysis, and obtainability of hybridization signals. RESULTS: We found that as archival age and tissue collagen content increased, not only did hydrolysis times have to be prolonged in order to yield interpretable hybridization signals, but also the likelihood of blocks becoming non-signaling increased. The most striking positive correlations were seen between the archival age of signaling lymphoid blocks and their requisite hydrolysis times. CONCLUSIONS: The difficulty in applying FISH on archival tissue increases with its archival age and collagen content, and may necessitate changes in laboratory protocol accordingly. PMID- 14529318 TI - Evaluation of temperature gradient capillary electrophoresis for detection of the Factor V Leiden mutation: coincident identification of a novel polymorphism in Factor V. AB - AIM: The Factor V Leiden mutation (G1691A) is a clinically important polymorphism that results in an increased risk of thrombosis. The goal of this study was to compare a temperature gradient capillary electrophoresis (TGCE) platform for the detection of Factor V gene mutations to a conventional restriction fragment length polymorphism (RFLP) assay. METHODS: Three hundred and four samples were analyzed by both TGCE and a common clinical Mnl I/RFLP assay. Concordance of results between the two assays was observed for 302/304 (99.3%) of the samples. RESULTS: All of the Leiden mutants (23/23, 100%) were identified by TGCE. Of the two discrepant results, one was caused by low peak heights in the TGCE output data and was easily rectified by the addition of a minimum peak height threshold. The second discrepancy resulted from the presence of a G-->A transition 95 bp downstream of the Leiden mutation site. This polymorphism represents a previously unreported alteration of the Factor V gene. CONCLUSIONS: The TGCE assay is less labor-intensive and has a higher throughput capacity than the Mnl I/RFLP assay. TGCE is a less specific assay than the Mnl I/RFLP assay that allows for the detection of novel polymorphisms, but also creates the need for all positive TGCE results to be confirmed by an alternate method such as sequencing. Our results demonstrate that TGCE is a highly sensitive method for mutation detection and has utility for mutation discovery analysis. PMID- 14529319 TI - Detection of Chlamydia pneumoniae in the cerebrospinal fluid of patients with multiple sclerosis by combination of cell culture and PCR: no evidence for possible association. AB - BACKGROUND: During the course of multiple sclerosis (MS) intrathecal oligoclonal IgGs are present in the cerebrospinal fluid (CSF). The intracellular human pathogen Chlamydia pneumoniae may play a role either as a causative pathogenetic agent in the disease, or C. pneumoniae-infected MS patients could be immunologically less able to clear the agent from the central nervous system (CNS). METHODS: CSF samples were studied in 100 individuals -- 70 MS patients and 30 age-matched controls with other neurological diseases. CSF was taken by lumbal puncture; cell cultures were performed by the cell vial technique, followed by a 4-day incubation at 37 degrees C. A nested PCR was performed. RESULTS: C. pneumoniae was detectable in the CSF of only 2.9% of the MS patients and none of control patients (with no significant difference between the MS patients and controls). IgG antibodies were positive in only 1.43% of the MS patients and 3.33% of the controls. IgA antibodies were positive in 6.66% of the control patients and none of the patients were positive for IgM antibodies. There was no statistically significant difference between the two groups of patients with respect to the three antibody classes. CONCLUSIONS: The results confirm the high leave of controversy surrounding a possible link between C. pneumoniae and MS, and the matter requires further thorough investigation. PMID- 14529320 TI - A one-step real-time PCR assay for rapid prenatal diagnosis of sickle cell disease and detection of maternal contamination. AB - INTRODUCTION: Mutations at the codon 6 of the beta-globin gene (hemoglobin [Hb] S and HbC) can be routinely identified by various methods and prenatal diagnosis has been available to affected families for several years. However, the presence of maternal cells in fetal samples constitutes a serious potential source of prenatal misdiagnosis and most methods currently used to detect maternal contamination are based on the analysis of highly polymorphic loci. In addition, these methods are labor intensive and time consuming and risk carry-over contamination. METHOD: We describe here a one-step method for mutation detection that uses fluorescent hybridization probes with melting curve analysis for both simultaneously prenatal diagnosis of sickle cell disease and potential maternal contamination. RESULTS: Retrospective and prospective prenatal diagnosis studies (conducted in 20 and 50 cases, respectively), using both the regular procedure and real-time PCR assay show perfect concordant results. We show in addition, that as little as 5% maternal contamination can be detected and that genotype determinations are unambiguous. PMID- 14529321 TI - Breast cancer: new technologies for risk assessment and diagnosis. AB - In the US, one in every eight women will develop breast cancer in her lifetime. Despite the advances made in treating breast cancer, the causal mechanisms underlying this disease have yet to be fully elucidated; 85% of breast cancer cases occur sporadically without any known genetic mutation. Too little is known about the pathogenesis of breast cancer for primary prevention to be feasible in the near- to mid-term. Secondary prevention through screening offers an alternative that has been widely adopted. For decades, breast self-examination has been touted as a technique for the early identification of breast cancer. However, it has been recently suggested that this technique is a waste of time and resources for both doctors and patients. Mammography finds breast cancer earlier than breast self-examination, and will reduce the risk of death from breast cancer by approximately 30% in women over 50 years old. Mammography is limited in that cancer, like breast tissue, appears white on the x-ray; therefore lesions may be difficult to detect in women with very dense breasts, and a tumor may not cast a significant shadow until it is quite large. Some cancers are so aggressive that they can spread quickly, before routine screening can detect them. Despite these limitations, mammography is still viewed as the best tool currently available for screening and early diagnosis. Improved methods to detect and diagnose breast cancer early, when it is most curable, are required if a significant impact on morbidity and mortality from breast cancer is to be made. Various new and innovative technologies are being investigated for improving the early detection and diagnosis of breast cancer. About 85% of breast cancers begin in the milk ductal system of the breast. As cancer develops in the breast, abnormalities occur, including atypical hyperplasia, ductal carcinoma in situ, and invasive breast carcinoma. Thus, the early screening of ductal cells can provide a parallel benefit to the 'Pap' smear, which is used virtually universally to identify the abnormal cells that can lead to cervical cancer. Two technologies to monitor for atypical ductal epithelial cells are Cytyc Corporation's FirstCyte Ductal Lavage system and Nastech Pharmaceutical Company's Mammary Aspiration Cytology Test. Matritech, Inc. is searching for biomarkers linked to breast cancer. Researchers at Matritech have detected the presence of nuclear matrix protein (NMP) in the blood of women at the early stage of breast cancer, which is absent in the blood of healthy women, as well as those with fibroadenoma, a benign breast disease. NMP66 has been selected as a marker for further development and clinical trials of a test for use in the detection and monitoring of women with, or at risk for, breast cancer have been initiated. Technologies developed by the US Department of Defense are under investigation as breast cancer screening. Advanced Image Enhancement, Inc. has licensed naval sonar technology for digital image enhancement of mammograms. New thermography applications are also being investigated in two separate projects sponsored by the US Department of Defense using military thermal surveillance tools adapted for cancer detection. Both are enhancements of older thermal imaging technology based on the principle that heat equates to unwanted activity, in the case of breast cancer, abnormal cell proliferation. PMID- 14529323 TI - Pharmacotherapy for attention-deficit/hyperactivity disorder (ADHD) decreases the risk for substance abuse: findings from a longitudinal follow-up of youths with and without ADHD. AB - Findings from a previously published longitudinal follow-up of adolescents with and without attention-deficit/hyperactivity disorder (ADHD) were reanalyzed to determine the extent to which pharmacotherapy for ADHD is associated with substance use disorders, including abuse and dependence. Using naturalistic data from 140 people with ADHD and 120 controls, researchers returned to previously accumulated data to determine whether exposure to stimulant therapy had increased rates of substance use disorder among ADHD patients. Findings included confirmation that, in fact, stimulant therapy protected medicated ADHD patients against substance use disorder, which occurred at rates that were 3 to 4 times greater among people with untreated ADHD. PMID- 14529324 TI - Does stimulant treatment lead to substance use disorders? AB - The authors examine the relationship between the treatment of attention deficit/hyperactivity disorder (ADHD) with stimulants and substance use disorders by reviewing their published meta-analysis of 6 studies and adding preliminary data from a seventh study. Despite some discrepancies among the findings of the 7 studies, the meta-analysis demonstrated that exposure to stimulant therapy for ADHD does not increase the risk for developing substance use disorders but is, in fact, protective against it. Stimulant treatment of ADHD appears to reduce the risk for substance use disorders by 50%, thus reducing the risk for substance use disorders in ADHD youth to levels well within the normal population risk. The implication of this finding is unquestionably one with enormous value both clinically and as a matter of public health. PMID- 14529325 TI - Comparing the abuse potential of methylphenidate versus other stimulants: a review of available evidence and relevance to the ADHD patient. AB - The use of psychostimulants to treat attention-deficit/hyperactivity disorder (ADHD) has been controversial for a number of reasons. In an effort to clarify the extent to which the psychostimulant methylphenidate has abuse potential, the existing published evidence has been reviewed and is summarized here, with an emphasis on delineating a number of related but independent issues that are often confused. Methylphenidate produces behavioral effects associated with abuse potential as assessed by traditional assays, but the relevance of this literature to the clinical use of the drug in the treatment of ADHD is ambiguous at best. Existing neuropharmacologic data suggest that methylphenidate has pharmacokinetic properties that reduce its abuse potential as compared with other stimulant drugs of abuse, such as cocaine. PMID- 14529326 TI - Childhood stimulant treatment and risk for later substance abuse. AB - Conflicting studies of the relationship between therapeutic use of psychostimulant medication and substance abuse have long been a subject of concern among clinicians and researchers. One controlled longitudinal study examined this relationship in 147 patients who were diagnosed with hyperactivity as young children and were surveyed with regard to their substance use both as adolescents and as adults. This study found that stimulant therapy for attention deficit/hyperactivity disorder in childhood is not associated with increased risk of adolescent experimentation with substance use, frequency of such use, or the risk of developing psychoactive substance use disorders by young adulthood. Moreover, stimulant therapy in high school may well have provided a protective effect against hallucinogen abuse by adulthood. A possible explanation for contradictory findings previously published was suggested by the existence of a number of potentially confounding variables, particularly conduct disorders, for which prior studies have failed to control. PMID- 14529329 TI - Judgement criteria for molar incisor hypomineralisation (MIH) in epidemiologic studies: a summary of the European meeting on MIH held in Athens, 2003. AB - AIM: This paper reviews the proceedings and conclusions of a meeting of experts concerned with the problem of hypomineralised incisors and molars (MIH). The aims of this meeting were to establish criteria for the judgement of MIH, to select representative cases and to discuss how the name of the condition was best described as hypomineralised, or hypomaturated, first permanent molars. CONCLUSION: There was not complete agreement as to the correct diagnosis and aetiology of the condition. A suggested list of definitions of the judgement criteria to be used in diagnosing Molar Incisor Hypomineralisation (MIH) for prevalence studies was agreed upon. PMID- 14529330 TI - Molar incisor hypomineralisation (MIH). AB - REVIEW: Molar Incisor Hypomineralisation (MIH) is defined as a hypomineralisation of systemic origin of one to four permanent first molars frequently associated with affected incisors. MIH molars are fragile and caries can develop very easily in those molars. Although MIH molars are well known by paediatric dentists and their occurrence is related in severe cases to major clinical problems, only limited data of the size of the problem are available. The prevalence of MIH ranges in the literature from about 3.6 to 25% and seems to differ in certain regions and birth cohorts. Unfortunately more complete comparable valid data are lacking at the moment. It seems that several aetiological factors can cause the enamel defects and that their occurrence is child related. CONCLUSION: For children with repeated illnesses in the first years after birth and children with opacities on erupted molars or incisors it seems useful to increase the frequency of dental check-ups during the period of erupting first permanent molars. PMID- 14529331 TI - Molar incisor hypomineralisation: restorative management. AB - AIM: The methods for the restoration of permanent molars and incisors affected by Molar Incisor Hypomineralisation are reviewed. The special problems associated with restoring these teeth, such as sensitivity, occlusion and aesthetics are discussed. The various options for restoration are outlined and recommendations made as to the appropriate types of restoration. PMID- 14529332 TI - Prevalence of enamel mineralisation disturbances in an area with 1-1.2 ppm F in drinking water. Review and summary of a report published in Sweden in 1981. AB - AIM: This was to study the prevalence of dental fluorosis in a Swedish city (Uppsala) with natural 1-1.2 ppm F in drinking water. It was also to compare the prevalence of idiopathic enamel disturbances (Morbus S) in children born in Uppsala with the prevalence in children who had moved in to Uppsala at an age when the Uppsala water could not have influenced the development of Morbus S. METHODS: Children born in 1970 and 1967 in Uppsala (Sweden) on even days (n=840) were invited to the study. All children not born in Uppsala were included (n=475), 24 children refused to take part and 45 were sick or had moved. Four examination teams performed the examinations after calibration against a "gold standard". Dental fluorosis was determined by the criteria of Dean [1934], idiopathic white spots according to Nevitt et al. [1963] and Morbus S was diagnosed according to Forsman [1979]. RESULTS: Children born in Uppsala had a higher prevalence and severity of fluorosis compared with those not born in Uppsala. It was found that 40 out of the 715 children born in Uppsala had enamel disturbances according to criteria for Morbus S and the corresponding figures for children not born in Uppsala were 33 out of 486. CONCLUSION: The specific enamel hypomineralisations type Morbus S cannot be connected to intake of fluoride. PMID- 14529333 TI - Hypomineralised first permanent molars and the orthodontist. AB - AIM: This paper discusses the orthodontic implications that need to be considered when a child presents with hypomineralised permanent molars. The various historical and present day orthodontic approaches are outlined and indications for the various sequences or extraction of first permanent molars presented. PMID- 14529334 TI - Molar incisor hypomineralisation in a group of children and adolescents living in Dresden (Germany). AB - AIM: This was to determine the prevalence of Molar Incisor Hypomineralisation (MIH) and to evaluate possible causes of these enamel defects. METHODS: The study group consisted of 2,408 10-17 year old children born during 1985-1992 and living in Dresden (Germany). Enamel defects were recorded using the modified DDE index. Children with enamel defects and their parents were invited for a re-examination to record the medical history of the children during the first 3 years of life; these children were matched by age with other children with apparently normal first molars (control group). RESULTS: 135 (5.6%) of the children had demarcated opacities in at least one first molar, i.e. MIH. A significantly higher prevalence of MIH was seen in children born between 1989 and 1991 compared with those born before and after that period (p<0.01). The number of children returning for the medical history questionnaire was low, 31 out of 135 responded (test group). Although there were no significant differences between the test and control groups in terms of peri and neonatal complications or other health problems, the low return precluded any definitive interpretation. CONCLUSION: The overall prevalence of MIH in this study was low by comparison with other previous epidemiological reports. PMID- 14529335 TI - A prospective clinical trial comparing preformed metal crowns and cast restorations for defective first permanent molars. AB - AIM: This study investigated two methods for the restoration of permanent molars affected by amelogenesis imperfecta (AI) or severe enamel defects. METHODS: A prospective clinical trial was carried out on 17 subjects affected by AI or severe enamel defects of first permanent molars. A split mouth design was used so that each right or left permanent molar in both jaws was restored using either a preformed metal crown (SSC) or a cast adhesive coping (CAC). Subjects were followed for up to 24 months and assessed for longevity and quality of the restorations. Sequential analysis was used to compare longevity. RESULTS: Records for 42 restorations (19 SSC; 23 CAC) were kept. The split mouth design was possible on 24 occasions (right versus left=14; maxilla versus mandible=10). Three restorations, one SSC (at 6 months) and two CAC (at 2 and 19 months) failed and required replacement. There was no significant statistical difference between the two types of restorations. CONCLUSION: While there was no difference between the two restorations for quality and longevity, the SSC was considerably cheaper to use and needed only one visit, but more tooth tissue was lost in preparation and fitting. The CAC was significantly more expensive but left nearly all of the tooth crown intact. The choice of which restoration to use is indicated by the immediate and long-term needs of each individual patient. PMID- 14529336 TI - Evaluation of composite restorations in hypomineralised permanent molars: a four year clinical study. AB - AIM: The objective of this investigation was to evaluate the clinical performance of composite restorations in enamel hypomineralised posterior teeth. METHODS: 52 composite restorations were placed in 52 permanent molars of 46 children, aged 8 10 years. All the teeth were clinically diagnosed as hypomineralised and restorations were placed on two or more surfaces of the teeth, including cusps. All treated teeth had at least 2 sound surfaces, thus excluding defective teeth with total disruption of the crown. The materials used were a hybrid composite and a fourth generation one-bottle adhesive material and manufacturer's instructions were carefully followed. The restorations were initially evaluated 7 days after the treatment and subsequent evaluation was performed at 12, 24, 36, 48 months. Clinical evaluation of the restorations was made according to the criteria of Ryge [1980]. RESULTS: In 6 cases, postoperative complaint was relieved after occlusal re-adjustment at the second appointment, 7 days later. At the end of the 48 months study period, 49 restorations were available for evaluation, all with full retention. Radiographically there was no periapical pathology. Problems were found in colour match in 10, surface appearance in 3 and anatomic form in 4 restorations. Hypersensitivity was recorded in 17 teeth after one week and in 3 teeth one year later. All teeth were sensitivity free after two years and until the end of the study period. CONCLUSIONS: Composite resin restorations using contemporary materials, in certain cases of hypomineralised permanent molars, can be an acceptable restorative procedure with satisfactory long-term results. PMID- 14529337 TI - Case report: clinical management of hypoplastic amelogenesis imperfecta. AB - BACKGROUND: The fundamental therapeutic problems related to amelogenesis imperfecta treatment are governed by the need to effect primary prevention interventions, reducing the risk of calculus accumulation and caries. There are also aesthetic and functional rehabilitative needs. Clinical management rehabilitation techniques vary depending on the AI type, but usually require restoration of affected teeth. Where orthodontic problems also exist, these should be corrected prior to the final restorative treatment. CASE REPORT: The use of composite resins in aesthetic restoration of permanent anterior and posterior teeth as affected by hypoplastic AI type is described. This case illustrates a patient who suffered from less attrition than those of the hypomineralized varieties. The therapeutic choice has allowed the correction of vertical dimension without the use of preformed crowns. PMID- 14529338 TI - Case report: orofacial characteristics of Hallermann-Streiff Syndrome. AB - BACKGROUND: Hallermann in 1948 and Streiff in 1950 described patients characterised by "bird face", congenital cataract, mandibular hypoplasia, and dental abnormalities. The new syndrome was later defined as Hallermann-Streiff Syndrome (HSS), underlining the differences with regard to Franceschetti's mandibulofacial dysostosis. CASE REPORT: Examination of a white male affected by Hallermann-Streiff Syndrome revealed facial characteristics typical of the "bird face" in HSS. The nose appeared thin, sharp and hooked; the prominence of the chin was absent in the lateral view; a marked microstomia was evident as well. Radiographic records showed aplasia of teeth 14, 13, 12, 22, 24, 35, 34, 33, 32, 31, 41, 42, and 43. As for orthodontic diagnosis, the following dentoskeletal features were assessed: skeletal Class II malocclusion, narrow upper arch, bilateral posterior crossbite, and anterior open bite. Lateral cephalogram showed hypoplasia of the mandible, a typical sign of HSS. The mandible revealed a clockwise rotation growth pattern with an opening of the gonial angle, skeletal anterior open bite, and an excessive vertical dimension of the lower third of the face. Radiographs at the age of 13 years showed an anomaly in shape of the lower right first bicuspid, which appeared with a double crown. TREATMENT: Orthodontic treatment started at 10 years of age with rapid maxillary expansion in the early stages of the mixed dentition, in order to correct the posterior crossbite due to the narrow maxillary arch. A subsequent phase of the orthodontic therapy consisted of a functional appliance with the goal of maintaining the transverse dimension of the dental arches and of controlling the tongue thrust in the anterior open bite. Surgical and prosthetic interventions were scheduled at completion of growth to solve the skeletal discrepancy and for occlusal rehabilitation. FOLLOW-UP: The therapeutic protocol used in the patient presented here was able to reach the orthodontic goals during the developmental ages. Further treatment is planned as there are multiple missing permanent teeth and prosthetic/restorative care is needed until the patient reaches full adulthood. PMID- 14529339 TI - An over four millennium story behind qinghaosu (artemisinin)--a fantastic antimalarial drug from a traditional chinese herb. AB - This article reviews the discovery and the further studies on qinghaosu and its derivatives or analogs over the last decades in the fields of medicinal chemistry and some pharmacology research, with an emphasis on the progress achieved in China. PMID- 14529340 TI - Progress in clinical, pharmacological, chemical and structural biological studies of huperzine A: a drug of traditional chinese medicine origin for the treatment of Alzheimer's disease. AB - HupA is a potent, reversible AChEI, which crosses the blood-brain barrier smoothly, and shows high specificity for AChE with a prolonged biological half life. It has been approved as the drug for the treatment of AD in China, and marketed in USA as a dietary supplement. HupA has been the subject of investigations by an ever-increasing number of researchers since 1980's. In the last four years, HupA has been further studied in many aspects such as the chemical synthesis, structural modification, structure-activity relationship, various biological effects, and mechanisms of action. A number of papers dealing with the computational modeling and X-ray crystallographic studies of HupA-AChE complex have also been published. This review represents a comprehensive documentation of the progress in the studies on HupA during the period of 1999 2002. PMID- 14529341 TI - The chemistry and biochemistry of Spiraea japonica complex. AB - The characteristic components of Spiraea japonica complex, which consists of seven varieties, are the hetisine- and atisine-type diterpene alkaloids, and the atisane-type diterpenes. From this complex, 20 hetisines, 37 atisines and 7 diterpenes were isolated during 1964-2001, including the observations of the isomerization of spiraea diterpene alkaloids having the oxazolidine ring and the interconversion of the oxazolidine rings, the configuration of the oxygen substitution at C-15 of the atisine-type alkaloids, and the interconversion relationship between the two main subtypes of the hetisine-type alkaloids. The chemotaxonomy based on structures of the reported diterpene alkaloids were suggested. The studies on the anti-inflammation, anti-platelet aggregation and neuroprotective bioactivities of the alkaloids from S. japonica var. acuta were reviewed as well. PMID- 14529342 TI - Natural product inspired design of enediyne prodrugs via rearrangement of an allylic double bond. AB - Cyclodeca-1,5-diyn-3-ene, the parent core of naturally occurring 10-membered ring enediyne antitumor antibiotics, decomposes at 37 degrees C with a t(1/2) value of 18 h. A prodrug approach was envisaged by in situ formation of the core structure via rearrangement of an allylic double bond. Three synthetic methods including intramolecular lithium acetylide addition to aldehyde, intramolecular Nozaki Hiyama-Kishi reaction, and intramolecular Sonogashira cross-coupling have been established for synthesis of the enediyne precursors, (E)-3-acyloxy-4 (arylmethylidene)cyclodeca-1,5-diynes. The latter are transformed into 10 membered ring enediynes in the presence of a catalytic amount of Ln(fod)(3) under mild reaction conditions. Alternatively, the enediyne precursor dissociates in buffer solution into an allylic cation or a quinone methide intermediate, which reacts with a nucleophile (such as H(2)O) to form the bioactive enediyne. LC-MS data confirmed formation of the 10-membered ring enediyne from the precursor, which exhibits DNA cleavage activity and cytotoxicity against P388 cancer cell line. PMID- 14529343 TI - Diversity oriented synthesis and branching reaction pathway to generate natural product-like compounds. AB - Combinatorial chemistry can be used to synthesize diversified molecules on a large scale. As with all large-scale experiments, this process requires a major investment in equipment, consumables and time. Therefore, careful design is critical. As the complexity of the libraries to be generated increases, additional considerations become important. What are the issues that should be considered when planning combinatorial chemistry projects? Which features in the design strategy are critical to consider ensuring that all of the potential products will be synthesized? How are the reactants selected to optimize product synthesis and yield? Over the last several years, through an experimental process, we have successfully developed and optimized our synthetic strategy. Our approach incorporates a number of critical components into a tightly controlled process that generates molecules with maximal structural complexity. This complexity emanates from carbon-carbon bond formation, which is extremely stable and it is reminiscent of complex natural product molecules. Our studies have illustrated that transition metal catalysts are powerful reagents that can be used to drive the synthesis of diverse small molecules from less complex starting materials. In this review, we will describe some of our recent efforts to synthesize natural product-like molecules and their derivative structures to successfully create libraries of complex molecules for drug discovery applications. Our diversity-oriented synthesis methods incorporate transition metal catalysts, as a versatile tool for creating carbon-carbon bonds and structural complexity, and the branched reaction pathway, as a method for incorporating diversity into the molecular scaffolds. We will review our combinatorial chemistry program, focusing on the decisions that we made for (1) the scaffold selection; (2) the design of a diversity oriented approach for library synthesis; (3) the incorporation of the branched reaction pathway to generate natural product-like molecules from the same starting material; and (4) the process steps that we selected for chemistry development and library generation. PMID- 14529344 TI - Enediyne natural products: biosynthesis and prospect towards engineering novel antitumor agents. AB - This review gives a brief account on the current status of enediyne biosynthesis and the prospective of applying combinatorial biosynthesis methods to the enediyne system for novel analog production. Methods for cloning enediyne biosynthetic gene clusters are first reviewed. A unified paradigm for enediyne biosynthesis, characterized with (a) the enediyne PKS, (b) the enediyne PKS accessory enzymes, and (c) tailoring enzymes, is then presented. Strategies and tools for novel enediyne analog production by combinatorial biosynthesis are finally discussed. The results set the stage to decipher the molecular mechanism for enediyne biosynthesis and lay the foundation to engineer novel enediynes by combinatorial biosynthesis for future endeavors. PMID- 14529345 TI - Virtual screening on natural products for discovering active compounds and target information. AB - Natural products, containing inherently large-scale structural diversity than synthetic compounds, have been the major resources of bioactive agents and will continually play as protagonists for discovering new drugs. However, how to access this diverse chemical space efficiently and effectively is an exciting challenge for medicinal chemists and pharmacologists. While virtual screening, which has shown a great promise in drug discovery, will play an important role in digging out lead (active) compounds from natural products. This review focuses on the strategy of virtual screening based on molecular docking and, with successful examples from our laboratory, illustrates the efficiency of virtual screening in discovering active compounds from natural products. On the other hand, the sequencing of the human genome and numerous pathogen genomes has resulted in an unprecedented opportunity for discovering potential new drug targets. Chemogenomics has appeared as a new technology to initiate target discovery by using active compounds as probes to characterize proteome functions. Natural products are the ideal probes for such research. Binding affinity fingerprint is a powerful chemogenomic descriptor to characterize both small molecules and pharmacologically relevant proteins. Therefore, this review also discusses binding affinity fingerprint strategy for identifying target information from the genomic data by using natural products as the probes. PMID- 14529346 TI - Stem cell gene therapy for chronic renal failure. AB - Recently, stem cell research has attracted considerable attention because it could be used for the regeneration of damaged organs that are untreatable by conventional techniques, and several stem cells (or progenitor cells), such as endothelial stem cells and neural stem cells have been discovered. Following the progression of this field of research, the potential for stem cell gene therapy has increased and several therapeutic benefits have already been reported. Although this approach was originally investigated for fatal or hereditary diseases, chronic renal failure is also a candidate for stem cell gene therapy. We have proposed two different therapeutic strategies for chronic renal failure depending on whether the bone marrow stem cells differentiate and commit into mesenchymal or hematopoietic stem cells. In the case of diseases, which need reconstitution of residential renal cells, such as congenital enzyme deficiency diseases, mesenchymal stem cells should be transplanted, and in contrast, hematopoietic stem cells may be used for gene delivery for diseases, which need foreign cytokines and growth factors, such as glomerulonephritis. This article reviews the recent investigation on this tailor-made stem cell gene therapy for chronic renal failure and discusses the potential of this novel strategy and the major practical challenges of its clinical application. PMID- 14529347 TI - Non-traditional drug targets: high risk, high reward. AB - Genomics and proteomics have unveiled a plethora of protein-protein interactions that may control cellular processes involved in disease development. Many of these interactions involve non-traditional candidate targets (i.e., neither enzymes nor cell surface receptors/channels). To date, non-traditional targets have largely been ignored by the pharmaceutical industry or have failed to lead to drugs. This review focuses on the use of transdominant genetically encoded agents and specialized small-molecule drugs to explore this void. PMID- 14529348 TI - Modified envelope glycoproteins to retarget retroviral vectors. AB - A conceptual breakthrough in gene therapy would be gene transfer vector that could be systemically applied, allowing targeted gene transfer into a predetermined cell type. The host range of a retroviral vector is determined by the interaction of the viral envelope glycoprotein (Env) and the retrovirus receptor on the surface of the host cell. In this review, we describe the current efforts to engineer targeted envelope glycoproteins, which can be incorporated into retroviral particles and are capable of delivering genes in a highly specific manner. PMID- 14529349 TI - Small interfering RNA: a new tool for gene therapy. AB - The use of anti-gene agents to disrupt the expression of disease-related genes could potentially be of utility in the treatment of a large number of illnesses, including most neoplasms. Traditional anti-gene agents include antisense oligonucleotides and ribozymes. Recent observations have provided evidence for another promising anti-gene technology-RNA interference (RNAi), in which the introduced double-stranded RNA (dsRNA), after a complicated series of processing steps, disrupts the expression of the targeted cellular gene. Further studies have indicated that small interfering RNAs (siRNAs) of generally 21 ~ 23 nucleotides, which resemble the processing products of long dsRNA, can induce RNAi directly in mammalian cells. Because of their high specificity and efficiency, siRNAs might be a new class of anti-gene medicines for gene therapy applications. PMID- 14529350 TI - Cardiac gene therapy: therapeutic potential and current progress. AB - Cardiovascular disease remains a major cause of morbidity and mortality in modern societies. While contemporary treatment modalities are making steady inroads to reduce this disease burden there remains a pressing need to vigorously explore novel therapeutic strategies. Rapid advances in our understanding of molecular pathology and the evolution of increasingly efficient gene transfer technology offer the imminent prospect of gene-based approaches to, at least, a subset of cardiovascular pathophysiologies. Initially envisaged as a treatment strategy for inherited monogenic disorders, it is now apparent that gene therapy has broader potential that encompasses acquired polygenic diseases, including many that affect the cardiovascular system. Extensive in vitro and animal studies are providing an increasingly sound scientific basis for cautious human evaluation. This review focuses on gene therapy of diseases primarily afflicting the heart, and provides an overview of gene and vector delivery systems with particular emphasis on systems suited to individual cardiac conditions. The pathophysiology underlying these conditions and molecular targets for therapeutic intervention are also reviewed. PMID- 14529351 TI - Integrated, molecular engineering approaches to develop prostate cancer gene therapy. AB - Gene therapy is a translational science, with the ultimate goal of cancer gene therapy research being to develop effective and safe treatments for patients. In the new millennium, it is imperative to tailor a therapeutic strategy for a particular disease, based on clinical management issues. The desirable regulatory features and therapeutic strategies need to be fully considered before proceeding with molecular engineering of the gene delivery vector. Issues, such as cell targeted expression, in vivo monitoring of gene delivery and expression, therapeutic strategies, and vector selection that targets the particular disease stage should be addressed. During the validation phase of the study, an objective evaluation in relevant animal models should determine whether the vector meets the desired specifications. Meeting the predetermined criteria should propel the product towards the clinical phase of evaluation. This review will present the conceptual framework that has been applied to developing an integrated and targeted gene therapy for prostate cancer. PMID- 14529352 TI - Receptor-mediated gene transfer vectors: progress towards genetic pharmaceuticals. AB - Although specific delivery to tissues and unique cell types in vivo has been demonstrated for many non-viral vectors, current methods are still inadequate for human applications, mainly because of limitations on their efficiencies. All the steps required for an efficient receptor-mediated gene transfer process may in principle be exploited to enhance targeted gene delivery. These steps are: DNA/vector binding, internalization, subcellular trafficking, vesicular escape, nuclear import, and unpacking either for transcription or other functions (i.e., antisense, RNA interference, etc.). The large variety of vector designs that are currently available, usually aimed at improving the efficiency of these steps, has complicated the evaluation of data obtained from specific derivatives of such vectors. The importance of the structure of the final vector and the consequences of design decisions at specific steps on the overall efficiency of the vector will be discussed in detail. We emphasize in this review that stability in serum and thus, proper bioavailability of vectors to their specific receptors may be the single greatest limiting factor on the overall gene transfer efficiency in vivo. We discuss current approaches to overcome the intrinsic instability of synthetic vectors in the blood. In this regard, a summary of the structural features of the vectors obtained from current protocols will be presented and their functional characteristics evaluated. Dissecting information on molecular conjugates obtained by such methodologies, when carefully evaluated, should provide important guidelines for the creation of effective, targeted and safe DNA therapeutics. PMID- 14529353 TI - Protein transduction domains and their utility in gene therapy. AB - Protein transduction domains (PTDs, sometimes termed cell permeable proteins (CPP) or membrane translocating sequences (MTS)) are small peptides that are able to ferry much larger molecules into cells independent of classical endocytosis. This property makes PTDs ideal tools to transfer proteins and other molecules into living cells for research purposes. The mechanism by which this internalization takes place is poorly understood. It is evident, however, that many known PTDs bind to the same surface molecules (Heparan Sulphate Proteoglycans, HSPG) before internalization, and that internalization is dependent on these molecules. PTDs, although at this moment mainly used for the chemical or bacterial production of membrane permeable proteins can become powerful tools for gene therapy. By incorporating a PTD in the therapeutic gene product, the protein produced in the transfected cell might be enabled to spread to non-transfected cells, thereby creating an increased therapeutic effect. In this review, we give an overview of PTDs that may be useful for gene therapy applications, and discuss some of the problems that can be expected when incorporating PTDs in gene therapy approaches. PMID- 14529354 TI - Targeting NS5B RNA-dependent RNA polymerase for anti-HCV chemotherapy. AB - The global prevalence of persistent hepatitis C virus (HCV) infection and the lack of a highly effective and well-tolerated antiviral therapy have spurred intensive efforts to discover and develop novel anti-HCV therapy in the pharmaceutical industry. HCV NS5B RNA-dependent RNA polymerase (RdRp), the centerpiece for viral replication, constitutes a valid target for drug discovery. Compared to the host RNA and DNA polymerases, NS5B RdRp has distinct subcellular localization at the interface of the endoplasmic reticulum (ER) membrane and cytoplasm, a novel catalytic mechanism and many unique structural features, all of which make it an attractive target for developing effective anti-HCV therapeutics. High genetic variation among the major HCV genotypes commands that any efficacious NS5B inhibitors have to be broadly active against NS5Bs from various genotypes. Rapid viral replication and its inherent genetic diversity will certainly culminate drug resistance to any NS5B inhibitors. Therefore, iterative drug design and combination therapies of drugs that intervene with different steps in the HCV replicative cycle are needed to combat the viral infection. Many classes of nucleoside and non-nucleoside inhibitors of NS5B RdRp have been identified and appeared in literatures and patent applications. These progresses hold a considerable promise to the development of novel, specific and highly effective therapeutics to achieve sustained response and ultimately the eradication of HCV infection. PMID- 14529355 TI - Molecular targets for papillomavirus therapy. AB - Papillomaviruses are infectious agents for human and animal epithelial tissue, and nearly 100 distinct human types (HPVs) have been identified. When these viruses infect cutaneous or mucosal skin they can initially cause clinical warts or persistent infection with little or no visible manifestations. Warts, while usually benign, can be painful or cosmetically unacceptable and often require medical treatment. Furthermore, infection with certain specific HPV types, such as 16 or 18 (as well as several others), is the major risk factor for a woman's development of cervical cancer. In addition to cervical cancer, papillomaviruses have also been implicated in cancers of the skin and respiratory track though the evidence is not yet as conclusive. It is clear that prevention or elimination of papillomavirus infections would ultimately reduce the incidence of cervical cancer and possibly other epithelial cancers as well. Unfortunately, progress in vaccine development has been slow and no specific anti-papillomavirus agents are available. The rational development of effective anti-papillomaviral treatments will require a detailed understanding of how these viruses replicate and interact with the host cell, and much progress has been made in this area over the last 10 years. These viruses have small DNA genomes with limited coding capacity, and their complete array of viral protein products is known. This review will discuss the known functions of the viral proteins with a focus on strategies to interdict their biological activities as a possible means of specific therapy. PMID- 14529357 TI - Current and future therapy for chronic hepatitis C virus liver disease. AB - Therapy with recombinant human interferon alpha remains pivotal to the treatment for chronic hepatitis C virus liver disease. Semi-synthetic protein-polymer conjugates of interferon with polyethylene glycol have also been recently developed. These conjugates protect the protein from degradation; reduce the immunogenicity; and prolong exposure to drug by a sustained absorption, restricted volume of distribution and sustained high serum concentration. Therapy with pegylated interferons is associated with significantly greater sustained virological response rates (SVR) compared to the non-pegylated formulation. Ribavirin is a guanosine analog with minimal antiviral activity against HCV. It demonstrates significant clinical synergism when administered in combination with interferon. Amantadine blocks entry of influenza A virus into cells. Used in combination with ribavirin and interferon as triple therapy, it may have some benefit compared to dual or monotherapy. Current treatment with pegylated interferons combined with weight-based ribavirin, provides the highest sustained virological response rates. In the absence of suitable animal models, HCV dynamic studies in man have been helpful in defining the mechanisms of action of interferon in chronic HCV liver disease. Novel therapeutic agents are being developed as the replication cycle of HCV is being understood. However, their safety and efficacy remain to be established and availability for clinical use is unlikely within the next 3 to 5 years. This review describes current antiviral therapy in chronic HCV liver disease, addresses the potential role of viral dynamics in elucidating the mechanisms of action of the drugs and discusses future potential therapeutics agents. PMID- 14529356 TI - Exploiting new potential targets for anti-hepatitis B virus drugs. AB - Based on the recent studies of HBV strains with different replication efficiency, several new potential targets for anti-HBV replication have been presented. These include the viral and cellular regulatory factors associated with HBV replication and the process for encapsidation of viral genome and budding into endoplasmic reticulum (ER). A putative regulatory domain has been reported at the carboxyl end of reverse transcriptase (RT) and when serine is substituted for proline at residue 652 of RT, replication efficiency of HBV is decreased. Substitution of proline for threonine at the 2798 nucleotide of the terminal protein (TP) gene, renders the mutant completely replication deficient. Expression of TP blocks the interferon (IFN) pathway and inhibits the responsive state of cells to interferons ( IFN) alpha and gamma. Interference of HBV capsid assembly drastically affects the encapsidation of viral genome, a crucial process for reverse transcription and viral DNA synthesis. Small molecules (bis-ANS) have been reported to act as a "wedge" to misdirect the polymerization of capsid, resulting in inhibition of virus replication. Another new group of compounds (HAP) has been shown to inhibit virus replication and also inhibit the assembly of viral capsid (core particle). Finally the capsids containing HBV genome are enveloped by budding into endoplasmic reticulum and release from virus infected cells, and this morphogenesis and secretion of HBV is dependent on glucosidases in the ER of host cells. Competitive inhibition of these glucosidases has been suggested as strategy against HBV replication. PMID- 14529358 TI - Vaccine development for potential bioterrorism agents. AB - Vaccines are considered to be one of the most effective ways of combating disease caused by bioterrorism agents. Such vaccines must be able to provide protection against pathogens which might enter the body by a number of routes, including the respiratory tract. They should also be able to induce protective immunity rapidly and would ideally be given non-invasively. There are few vaccines which currently meet these requirements. In part, this reflects the low level of research on many bioterrorism agents over the past few decades. Little is known about basic mechanisms of pathogenicity of many of these agents. However, by their very nature these agents cause serious disease, and must be handled in high containment laboratories. This requirement also limits the speed and ease with which research on these pathogens can now take place. Against this background, research on vaccines against potential bioterrorism agents is likely to proceed along two lines. Firstly because the genome sequences of most of the principal bioterrorism agents have either been completed or are close to completion, there is likely to be reliance on the exploitation of this information to devise improved vaccines. A number of groups are working on methodologies to identify vaccine antigens directly from genome sequences. Secondly, there will be a need to formulate such vaccines appropriately for the rapid induction of protective immunity after non-invasive delivery. The prospects for the development of a new generation of bioterrorism vaccines which exploit these technologies are reviewed in this manuscript. PMID- 14529359 TI - Vaccinia vectors as candidate vaccines: the development of modified vaccinia virus Ankara for antigen delivery. AB - Vaccinia viruses engineered to express foreign genes are powerful vectors for production of recombinant proteins. Originating from highly efficacious vaccines securing world-wide eradication of smallpox, the most appealing use of vaccinia vectors is to serve as vaccine delivery system for heterologous antigens. Concerns about the safety of vaccinia virus have been addressed by the development of vectors based on attenuated viruses. One of them, modified vaccinia virus Ankara (MVA) can be considered as current vaccinia virus strain of choice for clinical investigation. Historical development and use of MVA as vaccine against smallpox allowed to establish an extraordinary safety profile. MVA can be used under conditions of biosafety level 1 because of its avirulence and its deficiency to productively grow in human cells. In recent years significant progress has been made with regard to the development of MVA vector technologies. Compared to replication competent vaccinia viruses, MVA provides similar levels of recombinant gene expression even in nonpermissive cells. In animal models, MVA vaccines have been found immunogenic and protective against various infectious agents including immunodeficiency viruses, influenza, parainfluenza, measles virus, flaviviruses, or plasmodium parasites. By now first data from clinical trials are becoming available. In this article we briefly review history of MVA and state-of-the art technologies with regard to generation of recombinant MVA vaccines, and describe the progress to develop MVA vector vaccines against important infectious diseases. PMID- 14529360 TI - Poly(ADP-ribosyl)ation enzyme-1 as a target for neuroprotection in acute central nervous system injury. AB - Poly(ADP-ribose) polymerase 1 (PARP-1) protects the genome by functioning in the DNA damage surveillance network. In response to stresses that are toxic to the genome, PARP-1 activity increases substantially, an event that appears crucial for maintaining genomic integrity. Massive PARP-1 activation, however, can deplete the cell of NAD(+) and ATP, ultimately leading to energy failure and cell death. The discovery that cell death may be suppressed by PARP inhibitors or by deletion of the parp-1 gene has prompted a great deal of interest in the process of poly(ADP-ribosyl)ation. Suppression of PARP-1 is capable of protecting against cerebral and cardiac ischemia, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine induced parkinsonism, traumatic spinal cord injury, and streptozotocin-induced diabetes. The secondary damage of initially surviving neurons in brain stroke accounts for most of the volume of the infarcted area and the subsequent loss of brain function. Microglial migration is strongly controlled in living brain tissue by expression of the integrin CD11a, which is regulated in turn by PARP-1, proposing that PARP-1 downregulation may therefore be a promising strategy in protecting neurons from this secondary damage, as well. As PARP-1 is now recognised as playing a role also in the regulation of gene transcription, this further increases the intricacy of poly(ADP-ribosyl)ation in the control of cell homeostasis and challenges the notion that energy collapse is the sole mechanism by which poly(ADP-ribose) formation contributes to cell death. PARP(s) might regulate cell fate as essential modulators of death and survival transcriptional programs with relation to NF-kappaB and p53, proposing that inhibitors of poly(ADP-ribosyl)ation could therefore prevent the deleterious consequences of neuroinflammation by reducing NF-kappaB activity. PMID- 14529361 TI - The interleukin-1 system: an attractive and viable therapeutic target in neurodegenerative disease. AB - Inflammatory processes in the brain have been implicated in both acute and chronic neurodegenerative disease. One of the most studied inflammatory mediators in this respect is the cytokine interleukin-1 (IL-1), which has diverse actions in the central nervous system and mediates a wide variety of effects, including the host defense responses to local and systemic disease and injury. Both pre clinical and clinical data suggest a role for IL-1 as a key mediator of cell death in acute neurodegenerative conditions, such as stroke and head injury. IL-1 has also been implicated in a number of chronic diseases, including Parkinson's and Alzheimer's disease, as well as in epilepsy. Constitutive expression of IL-1 is very low in normal brain, but is up-regulated rapidly in response to local or peripheral insults. The mechanisms regulating the expression IL-1 are not well defined, but appear to involve multiple effects on neuronal, glial and endothelial cell function. Therefore, the IL-1 system represents an attractive and intensely competitive therapeutic target. PMID- 14529362 TI - Histamine H3 receptor: a potential drug target for the treatment of central nervous system disorders. AB - Histamine H(3) receptors were first described in the eighties but finally cloned four years ago. They are G-protein coupled, mostly presynaptic, and are involved in the control of the synthesis and/or release of different neurotransmitters both in the central nervous system and the periphery. The availabiliy of specific ligands has permitted the study of potential therapeutic applications of either stimulating or blocking the function of these receptors. There is experimental evidence that drugs targeted at histamine H(3) receptors could be beneficial for neurodegenerative diseases such as Alzheimer and Parkinson's disease, epilepsy, drug abuse and several affective, appetite and sleeping disorders, among others. This review presents recent advances in this field. PMID- 14529363 TI - Nerve growth factor: structure, function and therapeutic implications for Alzheimer's disease. AB - Over the past decade, neurotrophic factors have generated much excitement for their potential as therapy for neurological disorders. In this regard, nerve growth factor (NGF), the founding member of the neurotrophin family, has generated great interest as a potential target for the treatment of Alzheimer's disease (AD). This interest is based on the observation that cholinergic basal forebrain (CBF) neurons which provide the major source of cholinergic innervation to the cerebral cortex and hippocampus undergo selective and severe degeneration in advanced AD and that these neurons are dependent upon NGF and its receptors for their survival. In fact, NGF transduces its effects by binding two classes of cell surface receptors, TrkA and p75(NTR), both of which are produced by CBF neurons. This review focuses on NGF/receptor binding, signal transduction, regulation of specific cellular endpoints, and the potential use of NGF in AD. Alterations in NGF ligand and receptor expression at different stages of AD are summarized. Recent results suggest that cognitive deficits in early AD and mild cognitive impairment (MCI) are not associated with a cholinergic deficit. Thus, the earliest cognitive deficits in AD may involve brain changes other than simply cholinergic system dysfunction. Recent findings indicate an early defect in NGF receptor expression in CBF neurons; therefore treatments aimed at facilitating NGF actions may prove highly beneficial in counteracting the cholinergic dysfunction found in end-stage AD and attenuating the rate of degeneration of these cholinergic neurons. PMID- 14529364 TI - The endocannabinoid system and Huntington's disease. AB - The research in Huntington's disease (HD) has been growing exponentially during the last decade, since the discovery of the genetic basis that leads to neurodegeneration. HD is one of several progressive neurodegenerative disorders, in which the underlying mutation is a CAG expansion encoding a polyglutamine tract in a specific protein, which in the case of HD, is called huntingtin. The first clinical symptoms of HD are generally psychiatric abnormalities, most commonly depression and mood disturbances. Involuntary choreiform movements and dementia develop over the next 15-20 years, and death generally results from complications derived from immobility. There is currently no cure, or even an effective therapy to offset the decline in mental and motor capabilities suffered by those affected by HD, but recent studies have started to examine the usefulness of different classes of new compounds. Among these, plant-derived, synthetic or endogenous cannabinoids have been proposed to have therapeutic value for the treatment of HD, since they act on cannabinoid CB(1) receptors located in the basal ganglia circuitry, that is affected by the striatal atrophy typical of HD. Recent studies have characterized the changes in these receptors, as well as their endogenous ligands, in the basal ganglia in a variety of animal models of HD. The results are indicative that the endocannabinoid system becomes hypofunctional in this disease, which could be related to the hyperkinesia typical of the earliest phases of this disease. In addition, it has been proposed that the loss of these receptors might be involved in the process of pathogenesis itself. This, together with the well-known protective properties of cannabinoid related compounds, suggest that, in addition to a symptomatic usefulness, cannabinoids might also serve to delay or to arrest the development of this disease. The present article will review all recent data dealing with the biochemical, pharmacological and therapeutic bases that support a potential role of cannabinoids in the pathogenesis and/or therapeutic treatment of this motor disorder. PMID- 14529365 TI - On the estimation of binding affinity (deltaGbind) for human P450 substrates (based on Km and KD values). AB - A straightforward methodology, based on first principles, for the estimation of human cytochrome P450-substrate binding energies is outlined, and the system has then been applied successfully to a relatively large dataset of P450 substrates totalling 90 compounds. The results of Quantitative Structure-Activity Relationship (QSAR) analysis on the same dataset of cytochrome P450 (CYP) substrates from the CYP1, CYP2, and CYP3 families, involving a total of 90 compounds, agree favourably with the original analysis based on first principles, thus confirming the use of average values for hydrogen bond and pi-pi stacking energies, together with utilizing log P values as an estimation of desolvation energies. This method is based on a linear summation of the various contributary factors to the process, including: desolvation, hydrogen bonding, pi-pi stacking, restricted bond rotation and other energies relating to loss in translational and rotational energy. It is found that, for the majority of P450 substrates investigated, the first four terms are required for a relatively good estimation (R = 0.98) of the substrate binding affinity (deltaGbind) towards CYP1 and CYP2 enzymes. Consequently, it would appear that the loss in rotational and translational energy, which is thought to occur on substrate binding, apparently has little effect in most cases, possibly due to some degree of residual motion of the enzyme-substrate complex within the endoplasmic reticulum membrane. However, the appearance of a small constant term in the QSAR equation could possibly relate to an average loss in translational and rotational energy for the 90 compounds studied in this investigation. PMID- 14529366 TI - Atypical enzyme kinetics: their effect on in vitro-in vivo pharmacokinetic predictions and drug interactions. AB - The most common method for estimating a drug's in vivo clearance from in vitro data has involved using the classical Michaelis-Menten model to describe the observed in vitro data and from this estimate intrinsic clearance. This process assumes that the drug obeys standard enzyme kinetics that can be described by this model. However, the observation of atypical enzyme kinetics, particularly involving cytochrome P450 enzymes, has become relatively common, and occurrences have been reported with virtually all of the cytochrome P450 isoforms. Since predictions of a drug's in vivo clearance are made based on kinetic parameters observed during in vitro experiments (in vitro estimated intrinsic clearance), mis-identification of the drug's kinetic profile can lead to inaccurate predictions of intrinsic clearance. In addition to these potential ramifications of in vitro-in vivo predictions, information is becoming available concerning the in vivo implications of drugs that exhibit atypical kinetic profiles. This review will discuss the various types of atypical kinetic profiles that may be observed during in vitro experiments, hypotheses for why they may occur, examples of how a drug's intrinsic clearance may be mis-estimated when atypical kinetics are incorrectly assessed, and the potential ramifications of these issues for prediction of drug interactions. Furthermore, potential, artifactual causes of atypical kinetic phenomena will be described. PMID- 14529367 TI - Physiological approaches to the prediction of drug-drug interactions in study populations. AB - The prediction of metabolic drug-drug interactions should include quantitative attributes, such as variability in the study populations, and the results should be presented in terms of probability and uncertainty. The simple algebraic equations used to calculate one mean value for the extent of drug-drug interaction are adequate for qualitative or semi-quantitative risk assessment. However, truly quantitative predictions continue to fail. The success of drug drug interaction predictions requires understanding of the relationship between drug disposition and quantifiable influential factors on the change in systemic exposure. The complex interplay of influential factors, including variability estimates, on successful prediction of drug interaction have not been systematically examined. Therefore, physiologically relevant models of metabolic drug-drug interaction will likely play increasingly important roles in improving quantitative predictions and in the assessment of the influential factors underlying the interactions. The physiologically-based approach, with stochastic considerations, offers a powerful alternative to the empirical calculation of mean values. In addition to quantitative estimation of the interaction for assessing probability of risk, a reasonably validated predictive model is useful for prospective optimization of study designs. As a consequence, the definitive clinical trial would yield more meaningful information to support dosing recommendations. This review focuses on illustrating the importance of an integrated approach to building useful models for prediction of metabolism-based drug-drug interactions in human subjects. PMID- 14529368 TI - Application of a convective-dispersion model to predict in vivo hepatic clearance from in vitro measurements utilizing cryopreserved human hepatocytes. AB - Growing interest in the prediction of in vivo pharmacokinetic data from purely in vitro data has grown into a process known as the in vitro-in vivo correlation (IVIC). IVIC can be used to determine the viability of new chemical entities in the early drug development phases, leading to a reduction of resource spending by many large pharmaceutical companies. Here, a convective-dispersion model was developed to predict the total hepatic clearance of six drugs using pharmacokinetic data obtained from in vitro metabolism studies in which the drug disappearance from suspensions of human cryopreserved hepatocytes was measured. Predicted in vivo hepatic clearances estimated by the convective-dispersion model were ultimately compared to the actual clearance values and to in vivo hepatic clearances that were scaled based on the well-stirred model. Finally, sensitivity studies were performed to determine the dependence of hepatic clearance on a number of physiological model parameters. Results reaffirmed that low clearance drugs exhibit rate-limited metabolism, and their hepatic clearances are thus independent of blood flow characteristics, whereas drugs with relatively higher clearance values show a more pronounced dependence on the flood flow properties of dispersion and convection. Absent a priori knowledge about the flow-dependent properties of a drug's clearance, the convective dispersion model applied to disappearance data acquired from cryopreserved human hepatocytes is likely to provide satisfactory estimates of hepatic drug clearance. PMID- 14529369 TI - Relationship between extent of inhibition and inhibitor dose: literature evaluation based on the metabolism and transport drug interaction database. AB - A comprehensive search of the literature was undertaken using the Metabolism and Transport Drug Interaction Database (http://depts.washington.edu/didbase/) to evaluate the relationship between extent of inhibition and inhibitor dose. The search included reversible and irreversible inhibitors in studies conducted in the period 1966-2003. Only twelve inhibitors met the criterion of the search: study population exposed to more than one dose of inhibitor within a given study design. Six were reversible inhibitors: ciprofloxacin, enoxacin, felbamate, fluconazole, fluvoxamine and ketoconazole. The other six (cimetidine, diltiazem, disulfiram, paroxetine, verapamil and ritonavir) are considered irreversible inhibitors. Most of the AUC/Clearance data available for both types of inhibitors suggested evidence of dose-dependent inhibition. In the case of reversible inhibitors, the evidence of dose-dependent inhibition is consistent with a number of recent studies suggesting the determination of in vivo inhibition constants based on plasma concentration of inhibitor. PMID- 14529370 TI - Prediction of human drug metabolizing enzyme induction. AB - New chemical entities are routinely screened in vitro and in vivo for their ability to induce cytochrome P450s (CYP), other drug-metabolizing enzymes and possibly transporters in an attempt to more accurately predict clinical parameters such as drug-drug interactions and clearance in humans. Some of these potential therapeutic agents can cause induction of the metabolism of another molecule or auto-induction thereby increasing their own metabolism and elimination, as well as potentially any molecules metabolized by the same enzyme(s). Key CYPs in the 1A, 2B, 2C, and 3A families have all been shown to be inducible. It would be clearly advantageous to know the potential for a compound to induce drug metabolizing enzymes or transporters prior to clinical development, and many in vitro systems have been developed for this purpose. Newer computational technologies are also being applied in order to attempt to predict induction from the molecular structure alone before a molecule is even synthesized or tested. This review will cover the various in vitro and in silico methods developed for prediction of key inducers of CYPs and other proteins, as well as the limitations of such technologies and applications in the future. PMID- 14529371 TI - Transporter-enzyme interactions: implications for predicting drug-drug interactions from in vitro data. AB - As discussed in earlier articles, predictions of in vivo drug-drug interactions from in vitro studies is a subject of high interest with obvious therapeutic as well as economic benefits. Up until now little attention has been given to the potential interplay between metabolic enzymes and transporters that could confound the in vivo-in vitro relationships. Drug efflux by intestinal P glycoprotein (P-gp) is known to decrease the bioavailability of many CYP3A4 substrates. We have demonstrated that the interplay between P-gp and CYP3A4 at the apical intestinal membrane can increase the opportunity for drug metabolism by determining bidirectional extraction ratios across CYP3A4 transfected Caco-2 cells for two dual P-gp/CYP3A4 substrates, K77 (an experimental cysteine protease inhibitor) and sirolimus, as well as two negative control, CYP3A4 only substrates, midazolam and felodipine. Studies were carried out under control conditions, with a P-gp inhibitor (GG918) and with a dual inhibitor (cyclosporine). Measurement of intracellular concentration changes is an important component in calculating the extraction ratios. We hypothesize that the inverse orientation of P-gp and CYP3A4 in the liver will result in an opposite interactive effect in that organ. In vivo rat intestinal perfusion studies with K77 and rat liver perfusion studies with tacrolimus under control conditions and with inhibitors of CYP3A4 (troleandomycin), P-gp (GG918) and both CYP3A4/P-gp (cyclosporine) lend support to our hypotheses. These results serve as a template for predicting enzyme- transporter (both absorptive and efflux) interactions in the intestine and the liver. PMID- 14529372 TI - A new nonlinear mixture response surface paradigm for the study of synergism: a three drug example. AB - A flexible approach to response surface modeling for the study of the joint action of three active anticancer agents is used to model a complex pattern of synergism, additivity and antagonism in an in vitro cell growth assay. The method for determining a useful nonlinear response surface model depends upon a series of steps using appropriate scaling of drug concentrations and effects, raw data modeling, and hierarchical parameter modeling. The method is applied to a very large in vitro study of the combined effect of Trimetrexate (TMQ), LY309887 (LY), and Tomudex (TDX) on inhibition of cancer cell growth. The base model employed for modeling dose-response effect is the four parameter Hill equation [1]. In the hierarchical aspect of the final model, the base Hill model is treated as a function of the total amount of the three drug mixture and the Hill parameters, background B, dose for 50% effect D50, and slope m, are understood as functions of the three drug fractions. The parameters are modeled using the canonical mixture polynomials from the mixture experiment methodologies introduced by Scheff [2]. We label the model generated a Nonlinear Mixture Amount model with control observations, or zero amounts, an "NLMAZ" model. This modeling paradigm provides for the first time an effective statistical approach to modeling complex patterns of local synergism, additivity, and antagonism in the same data set, the possibility of including additional experimental components beyond those in the mixture, and the capability of modeling three or more drugs. PMID- 14529373 TI - A human drug metabolism database: potential roles in the quantitative predictions of drug metabolism and metabolism-related drug-drug interactions. AB - Previous attempts to predict drug metabolism and drug-drug interaction possibilities by deploying databases that house established drug metabolism results have been only marginally successful. Consideration of some of the key issues and concerns derived from these efforts suggests that three major hurdles loom in front of using xenobiotic metabolism databases more effectively in the future. These hurdles include: the need for an improved treatment of chemical structure in three-dimensions (3D); a better quantitative accounting of competitive and complementary biotransformation pathways; and, the critical need for a comprehensive, human drug metabolism database (hDMdb) that can serve as a bio/chemoinformatic resource pertaining to drug metabolism and drug metabolism related drug-drug interactions in general. Approaches that might be taken to traverse each of these hurdles are discussed herein where: the first involves maturation of chemical structures from simple 2D entries into more sophisticated 3D displays that can also account for interactions with relevant biological surfaces; the second involves a systematic, pair-wise comparison of various metabolic options in a statistically driven manner relative to chemical structure descriptors; and, the third involves mounting a hDMdb on the Internet in a user friendly manner that it is available via a non-profit format. PMID- 14529374 TI - Drug metabolism and drug interactions: application and clinical value of in vitro models. AB - In vitro models of drug metabolism are being increasingly applied in the drug discovery and development process as tools for predicting human pharmacokinetics and for the prediction of drug-drug interaction risks associated with new chemical entities. The use of in vitro predictive approaches offers several advantages including minimization of compound attrition during development, with associated cost and time savings, as well as minimization of human risk due to the rational design of clinical drug-drug interaction studies. This article reviews the principles underlying the various mathematical models used to scale in vitro drug metabolism data to predict in vivo clearance and the magnitude of drug-drug interactions resulting from reversible as well as mechanism-based metabolic inhibition. Examples illustrating the predictive utility of specific in vitro approaches are critically reviewed. Commonly encountered uncertainties and sources of bias and error in the in vitro determination of intrinsic clearance and metabolic inhibitory potency, including nonspecific microsomal binding, solvent effects on enzyme activities, and uncertainties in estimating enzyme available inhibitor concentrations are reviewed. In addition, the impact and clinical relevance of complexities such as dosing route-dependent effects, atypical multi-site kinetics of drug-metabolizing enzymes, non-cytochrome P450 determinants of metabolic clearance, and concurrent inhibition and induction, on the applicability and predictive accuracy of current in vitro models are discussed. PMID- 14529375 TI - Strategies for combinatorial biosynthesis with modular polyketide synthases. AB - Polyketides are assembled by the polyketide synthases (PKS) through a common mechanism, the condensation of small carboxylic acids. However, a large structural variety exists within these molecules, paralleled by their different bioactivities. Structural differences in polyketides mostly stem from variations in the number of elongation cycles, in the extender unit incorporated and the extent of processing occurring during each cycle. A significant fraction of polyketides is made in bacteria by modular PKSs, which direct polyketide synthesis on a protein template, where each module is responsible for selecting, incorporating and processing the appropriate carboxylate unit. Since their discovery in the early nineties, the architecture of modular PKSs and their modus operandi have attracted efforts by several laboratories to reprogram PKSs to produce tailor-made polyketides. The availability of a growing number of modular PKSs of defined sequence, and of well-developed model systems for the in vitro and in vivo analysis of these enzymes, has led to the successful production of many novel polyketides after genetic manipulation of the appropriate PKS. We discuss the different strategies that are followed for the construction of functional "hybrid" systems, with particular emphasis on what can be done in terms of generating chemical diversity, highlighting also the limitations of our current understanding. The prospects of generating novel useful polyketides by genetic engineering are also discussed. PMID- 14529376 TI - Expanding the scope of aromatic polyketides by combinatorial biosynthesis. AB - Combinatorial biosynthesis is a technology for mixing genes responsible for the biosynthesis of secondary metabolites, in order to generate products for compound libraries serendipitously or to cause desired modifications to natural products. Both of these approaches are extremely useful in drug discovery. Streptomyces and related species are abundant in bioactive secondary metabolites and were therefore the first microbes to be used for combinatorial biosynthesis. Polyketides are the most abundant medicinal agents among natural products. Structural diversity and a wide scope of bioactivities are typical of the group. However, the common feature of polyketides is a biosynthetic process from simple carboxylic acid residues. In molecular genetics, polyketides are sub-classified as types I and II, called modular and aromatic polyketides respectively. The best known bioactivities of aromatic polyketides are their antibacterial and antitumor effects. Genetic analysis of aromatic polyketides has resulted in almost 30 cloned and identified biosynthetic gene clusters. Several biosynthetic enzymes are flexible enough to allow their use in combinatorial biosynthesis to create high diversity compound libraries. This review describes the state of the art of combinatorial biosynthesis, giving anthracyclines as examples. Contiguous DNA sequences for antibiotics, cloned from four different anthracycline producers, provide tools for rapid lead optimization or other structural modification processes, and not only for anthracyclines. Two gene cassettes enabling fast and flexible structural modification of polyketides are introduced in this paper. PMID- 14529377 TI - On the generation of novel anticancer drugs by recombinant DNA technology: the use of combinatorial biosynthesis to produce novel drugs. AB - Chemotherapeutic drugs for cancer treatment have been traditionally originated by the isolation of natural products from different environmental niches, by chemical synthesis or by a combination of both approaches thus generating semisynthetic drugs. In the last years, a number of gene clusters from several antitumor biosynthetic pathways, mainly produced by actinomycetes and belonging to the polyketides family, are being characterized. Genetic manipulation of these antitumor biosynthetic pathways will offer in the near future an alternative for the generation of novel antitumor derivatives and thus complementing current methods for obtaining novel anticancer drugs. Novel antitumor derivatives have been produced by targetted gene disruption and heterologous expression of single (or a few) gene(s) in another hosts or by combining genes from different, but structurally related, biosynthetic pathways ("combinatorial biosynthesis"). These strategies take advantage from the "relaxed substrate specificity" that characterize secondary metabolism enzymes. PMID- 14529378 TI - Combinatorial biosynthesis of non-ribosomal peptides. AB - Non-ribosomal peptide synthetases (NRPS) are modular assembly lines catalysing the synthesis of many small peptides in microbes. Genetic replacements of domains or modules in NRPS encoded by gene clusters in Bacillus sp. with corresponding domains or modules from foreign NRPS have led in several cases to the in vivo synthesis of peptides with predicted amino acid substitutions. Fusion points were in variable regions between C- and A- or between T- and C-domains. Successful insertions of whole modules using fusion points in conserved regions internal to functional domains have also been reported. For studying the role of C-, A-, T- and TE (thioesterase)-domains in NRPS, several bi- and trimodular model-NRPS derived from natural NRPS systems were constructed and obtained after expression in E. coli with coexpression of a 4'- phosphopantetheine transferase or in suitable hosts such as the Streptomyces. Such enzymes were shown to catalyse in vitro synthesis of di- and tripeptides, respectively, with and without turnover depending on the presence of Te-domains. The enzymatic analysis revealed the mechanisms of the domains and proved their functional autonomy suggesting the possibility to use any NRPS interdomain region for fusions. Nevertheless, recombinant synthesis of longer and more complex peptides will still be restricted to alteration of existing structures by manipulations of NRPS gene clusters located on chromosomes or artificial chromosomes. Besides targeted replacements of domains and modules, reprogramming of NRPS by altering the substrate specificities of A-domains is a promising tool for the future to get novel peptides. PMID- 14529379 TI - Diversity among microbial cyclic lipopeptides: iturins and surfactins. Activity structure relationships to design new bioactive agents. AB - A prominent group of bioactive lipopeptides produced by Bacillus species is constituted by iturins, surfactins and lichenysins. Interest in such substances results in their exceptional surfactant power, and their valuable antifungal, antibacterial, antitumoral and anti-Mycoplasma properties. As is typical for peptidic secondary-metabolites synthesized by the polyenzymic pathway, they are produced as mixtures of components varying in the peptidic and/or in the lipidic structure. In the context of structure-activity relationships, it is possible to take advantage of the adaptability of the biosynthesis system by systematically adding selected amino acids in the culture medium of the producing bacterium. When an amino acid is used as the sole nitrogen source, it is inserted directly into selected positions of the peptide sequence, thus amplifying the original structural microheterogeneity via a production of variants. This method revealed very efficient for increasing the amounts of preexisting variants and for building new variants of surfactins and lichenysins but totally inefficient with iturins. In this group, the peptidic diversity strictly depends on the selected strain. So far the screening remained the only method to discover new iturins. Another interesting peculiarity is the common occurrence in a single strain of two lipopeptides with different core structures such as surfactins and iturins. Taken together, these features led to an extensive metabolite pattern. Besides, engineered variants and chemical derivatives enlarged the array of available molecules. Despite the high degree of chemical similarity, the separation of variants and/or homologues was successfully achieved by reversed-phase HPLC leading to well-separated compounds ideally suited to investigation of structure activity relationships. Improved physical techniques such as 2D-NMR and mass spectrometry allowed to describe efficiently and rapidly the composition of cyclic lipopeptides even in mixtures containing several variants. From NMR, the 3D structure and dynamics gave crucial data for fine structure-activity relationships as well as for understanding of the properties at the membrane and/or at the air/water interface. Here the role of residues was identified in the context of hydrophobic and electrostatic interactions that play a leader role. Such a comprehensive approach, based on both structural and biosynthesis knowledge, opened the way to rational design for enhanced properties and its validity was confirmed with 10 fold higher surfactant efficacy. PMID- 14529380 TI - "Whole cell"--matrix-assisted laser desorption ionization-time of flight-mass spectrometry, an emerging technique for efficient screening of biocombinatorial libraries of natural compounds-present state of research. AB - Whole Cell-matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) is an emerging sensitive technique for rapid typing of microorganisms, efficient screening of biocombinatorial libraries of natural compounds and the analysis of complex biological samples, as whole cells, subcellular particles, cell extracts and culture filtrates. It is unique to detect metabolites in-situ without the need to isolate and purify the investigated compounds. In favourite cases it enables in-situ structure analysis on the basis of the fragment pattern generated by postsource MALDI-TOF-mass spectrometry. The state of research of this methodology which has mainly been obtained by investigation of lipopeptides from bacilli and the large spectrum of bioactive peptides produced by cyanobacteria is reviewed. The potential of this innovative technique is demonstrated for the lipopeptides produced by various Bacillus subtilis strains. PMID- 14529381 TI - Combinatorial chemistry in food research. AB - This mini-review is concerned with emerging applications of combinatorial chemistry relevant to the needs of the food industry. More specifically, recent advances in the use of combinatorial methods for the identification and analysis of flavours, "food chemical" libraries as a potential source of enzyme inhibitors for the food industry and the utility of biocatalysis for the generation of molecular diversity are discussed. PMID- 14529382 TI - Overview: translating Hsp90 biology into Hsp90 drugs. AB - The Hsp90 molecular chaperone has emerged as one of the most exciting targets for cancer drug development. Hsp90 is overexpressed in many malignancies, very likely as a result of the stress that is induced both by the hostile cancer microenvironment and also by the mutation and abberant expression of oncoproteins. A particularly attractive feature of Hsp90 as a cancer drug target is that it is required for the conformational stability and function of a wide range of oncogenic 'client' proteins, including c-Raf-1, Cdk4, ErbB2, mutant p53, c-Met, Polo-1 and telomerase hTERT. Inhibition of Hsp90 should therefore block multiple mission critical oncogenic pathways in the cancer cell, leading to inhibition of all the hallmark traits of malignancy. This combinatorial blockade of oncogenic targets should give rise to board spectrum antitumour activity across multiple cancer types. The 'druggability' of Hsp90 was confirmed by the discovery that the natural products geldanamycin and radicicol, which have anticancer activity, exert their biological effects by inhibiting the essential ATPase activity associated with the N-terminal domain of the protein. The first in-class Hsp90 inhibitor has entered clinical trial and provided proof of concept that Hsp90 can be inhibited and clinical benefit seen at non-toxic doses. Further development is underway and a related analogue 17DMAG also shows promise in preclinical models. In addition, novel Hsp90 inhibitors have been identified using methods such as high throughput screening and x-ray crystallography. The opportunities and challenges involved in translating the fast moving biology of Hsp90 into patient benefit is discussed. PMID- 14529383 TI - Structure and functional relationships of Hsp90. AB - Understanding the mode of action of Hsp90 requires that molecular detail of its interactions with client proteins and co-chaperones are known. The structure determination of the N-terminal domain of Hsp90/Hsp90beta, proof that it is an ATPase, that this activity is regulated and the identification of co-chaperones that facilitate Hsp90 function were landmarks towards understanding conformational changes in Hsp90 brought about by ATP, co-chaperones and client proteins. Sti1 and Cdc37/p50, which associate with early Hsp90 complexes, were shown to be inhibitors of Hsp90 ATPase activity and therefore promote its 'open' state, whereas Sba1/p23, which associates with mature complexes, inhibits ATPase activity and stabilises the 'closed' state. The isolation and characterisation of Aha1, the only known strong activator of Hsp90 ATPase activity, which promotes the 'closed' state of Hsp90, will also be of major importance in understanding Hsp90 function. The structure determination of the middle region of Hsp90 has shed further light on the complex ATP-cycle of Hsp90, identifying a catalytic loop, with key residues that are essential for ATP hydrolysis. These studies, together with biochemical ones, suggest that ATP hydrolysis, is dependent on a complex rate-limiting step, involving N-terminal dimerization and association of the middle region, and therefore the catalytic loop, of Hsp90 with the N-terminal domains. The structure of the middle region of Hsp90 will also accelerate our understanding of client protein interactions since this region is implicated in their recognition and in particular their active-site openings. PMID- 14529385 TI - Genes and proteins governing the cellular sensitivity to HSP90 inhibitors: a mechanistic perspective. AB - HSP90 inhibitors such as 17AAG have the major therapeutic advantage that they exert downstream inhibitory effects on multiple oncogenic client proteins. They therefore block several mission critical cancer-causing pathways and have the potential to modulate all of the hallmark biological features of malignancy. Consistent with this combinatorial anti-oncogenic profile, 17AAG exhibits broad spectrum antitumour activity against cultured cancer cell lines and in vivo animal models. However, there are clear differences in sensitivity between various cancer cell lines and it is quite possible that some tumour types or individual patients will be more responsive in the clinic than others. We describe the methods used to investigate the genes and proteins involved in the mechanism of action of HSP90 inhibitors and discuss the significance of these for cellular sensitivity. Methods used involve the conventional cell and molecular biology techniques, together with the more recent application of high throughput global technologies such as gene expression microarrays and proteomics. Selected examples that seem to play a role in sensitivity to HSP90 inhibitors are highlighted and the potential relevance to the response of cancer patients is discussed. Important determinants of response include: 1) Dependence upon key HSP90 client proteins such as ERBB2, steroid hormone receptors and AKT/PKB; 2) Levels of HSP90 family members and co-chaperones, such as HSP70 and AHA1; and 3) expression of various cell cycle and apoptotic regulators. In the case of 17AAG, metabolic enzymes such as NQO1 and membrane efflux pumps are also important for sensitivity. PMID- 14529384 TI - Natural product origins of Hsp90 inhibitors. AB - The currently used Hsp90 inhibitors, geldanamycin, herbimycin A and radicicol, were isolated many years ago from Streptomyces and fungi originally for their antiprotozoal activity, herbicidal activity and antifungal activity, respectively. In the mid 1980s, it was found that the benzoquinone ansamycin antibiotics (herbimycin A, geldanamycin, and macbecin) reversed v-Src transformed cells to normal phenotypes, and Bcr-abl was subsequently suggested to be the molecular target for the treatment of chronic myelogenous leukemia through a study using herbimycin A for its selective antioncogenic activity. In 1994, these ansamycins were found to bind to Hsp90 and to cause the degradation of client proteins including Src kinases; further efforts to develop anticancer drugs were made using geldanamycin analogs, and 17AAG was chosen as the best candidate for clinical trials. The number of novel natural products isolated from microbial origins is continuing to increase and is doubling every 10 years. Thus, screening of bioactive substances from natural origins, using assays including defined targets, and developing leads toward drugs via optimized derivatization is a conventional but still promising strategy for drug discovery and development. PMID- 14529386 TI - The stress response: implications for the clinical development of hsp90 inhibitors. AB - In their role as molecular chaperones, heat shock proteins serve as central integrators of protein homeostasis within cells. As part of this function, they guide the folding, assembly, intracellular disposition and proteolytic turnover of many key regulators of cell growth, differentiation and survival. Not surprisingly then, heat shock proteins are over expressed in many types of cancer, and induction of the stress response may actually be required for cells to tolerate the genetic disarray characteristic of malignant transformation. Regulation of heat shock protein levels via the stress response is complex, but recent data indicate that the molecular chaperone Hsp90 plays a key role. Specifically, Hsp90 inhibitors alter the multi-chaperone complexes associated with Heat Shock Factor 1 (HSF1), the dominant transcription factor controlling induction of the stress response, and stimulate HSF1-activated heat shock gene expression. Induction of this heat shock response has now emerged as an important consideration in the further clinical development of Hsp90 inhibitors for several reasons. First, tumors in which the stress response is compromised appear particularly sensitive to Hsp90 inhibition. Second, induction of the stress response by Hsp90 inhibitors provides a sensitive pharmacodynamic endpoint with which to monitor drug action in individual patients. Third, Hsp90 inhibitors display important therapeutic interactions with both conventional DNA-targeted chemotherapeutics and newer molecularly targeted agents. These interactions are, at least in part, due to modulation of the stress response by these drugs. Lastly, stress response induction by Hsp90 inhibitors may have therapeutic benefits in non-neoplastic disorders such as heart disease, stroke and neurodegenerative diseases. These benefits are just beginning to be explored. PMID- 14529387 TI - Development of radicicol analogues. AB - Radicicol, a macrocyclic antibiotic produced by fungi, was originally isolated many years ago, and was described as tyrosine kinase inhibitor. We also rediscovered radicicol as an inhibitor of signal transduction of oncogene products, such as K-ras and v-Src, using yeast and mammalian cell-based assays. In a study of mechanisms of action, it was revealed that radicicol depletes the Hsp90 client signaling molecules in cells, and thus inhibit the signal transduction pathway. In addition, direct binding of radicicol to the N-terminal ATP/ADP binding site of Hsp90 was shown, and thus radicicol has been recognized as a structurally unique antibiotic that binds and inhibits the molecular chaperone Hsp90. Although radicicol itself has little or no activity in animals because of instability in animals, its oxime derivatives showed potent antitumor activities against human tumor xenograft models. Hsp90 client proteins were depleted and apoptosis was induced in the tumor specimen treated with radicicol oxime derivatives. Taken together, these results suggest that the antitumor activity of radicicol oxime derivatives is mediated by binding to Hsp90 and destabilization of Hsp90 client proteins in the tumor. Among Hsp90 clients, we focused on ErbB2 and Bcr-Abl as examples of important targets of Hsp90 inhibitors. Radicicol oxime showed potent antitumor activity against ER negative/ErbB2 overexpressing breast cancer and Bcr-Abl expressing CML. Putative mechanisms of action and future directions of radicicol oxime against these kinds of tumor are discussed. PMID- 14529388 TI - Development of purine-scaffold small molecule inhibitors of Hsp90. AB - The Hsp90 chaperones play a key role in regulating the physiology of cells exposed to environmental stress and in maintaining the malignant phenotype in tumor cells. Agents that interfere with the function of the chaperone may thus be beneficial in the treatment of cancers. The ansamycins (geldanamycin and herbimycin) and the unrelated natural product radicicol were found to bind to the N-terminal pocket of Hsp90 and inhibit its function. However, translation of these compounds to the clinic was impeded by stability and hepatoxicity issues. 17AAG, a derivative of geldanamycin, was found to be less hepatotoxic and is currently undergoing Phase I clinical trial. Unfortunately, 17AAG is insoluble, difficult to formulate and it is not yet clear if therapeutically effective doses can be administered without escalating non-Hsp90 associated toxicities. Additionally, for reasons not yet completely understood, a subset of tumor cells are insensitive to the action of the drug. The development of novel agents that lack the drawbacks of the natural products is thus necessary. Here we present an overview of such efforts with focus on a new class of purine-scaffold Hsp90 inhibitors developed by rational design. PMID- 14529389 TI - Clinical development of 17-allylamino, 17-demethoxygeldanamycin. AB - 17-allylamino, 17-demethoxygeldanamycin (17AAG; NSC 330507) is the first modulator of heat shock protein 90 (Hsp90) to enter clinical trials. Hsp90 serves a chaperone role to properly fold and deliver client proteins to appropriate intracellular locations. Interest in Hsp90 modulators for the experimental therapeutics of cancer has arisen based on pre-clinical evaluations suggesting that Hsp90 client proteins regulate signaling pathways critical to the molecular economy of many types of tumors, including oncogene signaling, cyclin-dependent kinase activation, steroid hormone receptors, and mediators of invasion and metastasis. Thus, Hsp90-directed agents could affect molecules upon which tumors depend for their proliferation and survival. Initial clinical studies have therefore sought to incorporate assessment of these endpoints into initial clinical evaluations. Three schedules of administration have been supported for initial evaluation in Phase I studies sponsored by the National Cancer Institute (NCI) or supported by NCI and sponsored by Cancer Research UK. In the daily times five schedule, a recommended Phase II dose (RPTD) of 40 mg/m(2) has been reached, while once weekly or three of four weekly schedules are defining RPTDs of 295 and 308 mg/m(2). Toxicity is tolerable and appears dominated by hepatic, gastrointestinal, and constitutional symptoms. Concentrations of drug at peak of ~1700-3000 nM are concordant with concentrations predictive of useful outcomes in pre-clinical model systems. Evidence of modulation of Hsp90 partner molecules has been obtained in both surrogate and some tumor compartments. These very early results encourage additional clinical evaluations of 17AAG and related molecules. PMID- 14529390 TI - The clinical applications of heat shock protein inhibitors in cancer - present and future. AB - The potential clinical applications of the prototype first-in-class Hsp90 inhibitor 17AAG and other emerging Hsp90 drugs are very exciting. Rigorously planned and executed clinical trials, incorporating measurement of appropriate biomarkers and pharmocodynamic endpoints are critical for selecting the optimal dose and schedule. A detailed understanding of the molecular mode of action of Hsp90 inhibitors alongside the elucidation of the molecular pathology of individual cancers will help us to identify tumour types and individual patients that will benefit most from treatment. Careful in vitro and in vivo experiments are needed to choose the most potentially advantageous combination studies. It is important to construct a pharmacologic audit trail linking molecular biomarkers and pharmacokinetic and pharmacodynamic parameters to tumour response endpoints. Phase I clinical studies with 17AAG have shown that the drug can be given at does that are well tolerated and that also achieve active pharmacokinetic exposures and elicit molecular signatures of gene and protein expression that are indicative of Hsp90 inhibition. Furthermore, examples of disease stabilisation have been documented, consistent with the generally cytostatic responses that are seen in animal models. Selecting tumour types for Phase II clinical trials must involve balancing 1) our knowledge of molecular response determinants, such as the expression of and dependence upon key client proteins and 2) more pragmatic evidence of antitumour activity in the relevant preclinical models. Examples of likely disease targets include chronic myeloid leukaemia, melanoma, breast, ovarian, brain, thyroid, colorectal and prostate cancer. PMID- 14529391 TI - Cilostazol as a unique antithrombotic agent. AB - Cilostazol (CLZ) was originally developed as a selective inhibitor of cyclic nucleotide phosphodiesterase 3 (PDE3). PDE3 inhibition in platelets and vascular smooth muscle cells (VSMC) was expected to provide an antiplatelet effect and vasodilation. Recent preclinical studies have demonstrated that CLZ also possesses the ability to inhibit adenosine uptake by various cells, a property that distinguishes CLZ from other PDE3 inhibitors, such as milrinone. After extensive preclinical and clinical studies, CLZ has been shown to have unique antithrombotic and vasodilatory properties based upon these novel mechanisms of action. CLZ was approved in 1988 for the treatment of symptoms related to peripheral arterial occlusive disease in Japan (Pletaal) and in 1999 in the U.S. and in 2001 in the U.K. (Pletal) for the treatment of intermittent claudication symptoms. Despite its remarkable antiplatelet properties, CLZ is not generally considered an antithrombotic agent in Western countries, perhaps due to the bulk of its antithrombotic preclinical and clinical development being conducted in Japan. In this review, the unique properties of CLZ are reviewed with the focus on CLZ as a unique antiplatelet agent targeting platelets and VSMC, demonstrating synergy with endogenous mediators and showing lowered risk of bleeding risk compared to other antiplatelet drugs. PMID- 14529392 TI - ADP receptors--targets for developing antithrombotic agents. AB - Platelet P2 receptors--P2Y1, P2Y12, and P2X1--constitute the means by which adenine nucleotides can activate platelets. Coactivation of the Galphaq-coupled P2Y1 and Galphai2-coupled P2Y12 receptors is necessary for ADP-mediated platelet activation, which forms the basis of using P2 antagonists as antithrombotic drugs. P2Y1 receptor antagonists inhibit platelet activation, while P2Y1 knockout mice show longer bleeding times than normal mice but few other problems; however, its ubiquitous expression in other tissues renders P2Y1 questionable as an antithrombotic target. The P2Y12 receptor is expressed nearly exclusively in platelets and brain, making it an attractive antithrombotic target. Antagonists for the P2Y12 receptor have been developed that either require metabolic activation to covalently inhibit P2Y12 and are irreversible, or simply are competitive in nature and thus reversible. Ticlopidine and clopidogrel are irreversible P2Y12 antagonists and have been repeatedly proven as clinical antithrombotic agents. In addition, a recently reported P2Y12 antagonist, CS-747, shows promise as a future antithrombotic drug. The AR-C series of compounds represent reversible P2Y12 antagonists and have been used extensively to characterize the function of P2Y12 in platelets. Clinical studies show that AR C69931MX is as effective as clopidogrel; furthermore, the combination of AR C69931MX (cangrelor) and clopidogrel confers greater antagonism of P2Y12 than either antagonist alone. The P2X1 receptor is a calcium channel that functions to potentiate agonist-induced platelet shape change, and its inhibition or loss has little if any effect on hemostasis. A combination of P2Y1 and P2Y12 antagonists may represent an additional course of antithrombotic treatment. PMID- 14529393 TI - Antiplatelet therapies: platelet GPIIb/IIIa antagonists and beyond. AB - Cardiovascular and cerebrovascular diseases continue to be leading causes of death throughout the world. Blood platelets play a pivotal role not only in haemostasis but also in the pathogenesis of thrombosis and atherosclerosis, platelet aggregation being an essential step in the formation of either an effective haemostatic plug or an intravascular thrombus. The benefits of various antiplatelet therapies ranging from aspirin, ticlopidine, Clopidogrel, and intravenous platelet GPIIb/IIIa antagonists in various thromboembolic disorders are well documented. Despite of the success of intravenous acute GPIIb/IIIa blockade when given in conjunction with heparin, chronic oral GPIIb/IIIa antagonists with or without aspirin failed in various cardiovascular settings. This review highlights the role of the various antiplatelet therapies in thrombotic disorders as well as future directions. PMID- 14529394 TI - Synthetic oligosaccharides as heparin-mimetics displaying anticoagulant properties. AB - Heparin and low molecular weight heparins are major clinical anticoagulants and the drugs of choice for the treatment of deep venous thrombosis. The discovery of an antithrombin binding domain in heparin focused interest on understanding the mechanism of heparin's antithrombotic/ anticoagulant activity. Various heparin mimetic oligosaccharides have been prepared in an effort to replace polydisperse heparin and low molecular weight heparins with a structurally-defined anticoagulant. The goal of attaining a heparin-mimetic with no unwanted side effects has also provided motivation for these efforts. This article reviews structure-activity relationship (SAR) of structurally-defined heparin-mimetic oligosaccharides. PMID- 14529395 TI - Recent advances in the discovery and development of direct coagulation factor Xa inhibitors. AB - Coronary heart disease (CHD) is the leading cause of mortality and morbidity in the United States. Currently, there are approximately 12 million Americans with CHD, which is most frequently caused by atherosclerosis. The thrombotic complications of atherosclerosis, such as acute coronary syndrome and ischemic stroke, can be fatal and those who survive such events have a far greater risk of future cardiovascular events. This huge medical need cries out for improved novel anticoagulants, antiplatelet agents, and profibrinolytic agents. These agents will successfully respond to the medical need by providing safe, effective, and easily administered treatments that have little, if any, drug and food interactions and that require minimal monitoring. The currently approved antiplatelet agent, clopidogrel, has satisfied some of these requirements and has played a large role in expanding the antithrombotic market over the past few years. New antithrombotics approaching the marketplace, such as the prodrug thrombin inhibitor ximelagatran, have promise in expanding the antithrombotic market further. Over the past two decades, the pharmaceutical industry has mounted a huge effort to develop antithrombotics that function by inhibiting key enzymes positioned at "higher" levels of the coagulation system. Direct inhibitors of factor Xa, which may provide a better safety and efficacy profile than currently available agents, appear to be the next major class of antithrombotic agents poised to take the pharmaceutical industry one step closer to delivering the ideal antithrombotic agent. This review focuses on recent innovations in the discovery and development of potent parenteral and oral direct factor Xa inhibitors. PMID- 14529396 TI - Development of proteinase-activated receptor 1 antagonists as therapeutic agents for thrombosis, restenosis and inflammatory diseases. AB - Thrombin, a plasma serine protease, plays a key role not only in coagulation and hemostasis but in thrombosis, restenosis and atherosclerosis. Thrombin activates platelets, endothelium, inflammatory cells and smooth muscle cells. The cellular action of thrombin is mediated by specific G-protein coupled thrombin receptors called proteinase-activated receptors (protease-activated receptor or PARs). Among the three thrombin receptors, PAR1 is the primary thrombin receptor in human and animal cells with an exception of non-primate platelets. An increased thrombin generation and PAR1 expression are observed on cells within atherosclerotic plaque and thrombus and following vascular injury. Animal studies with PAR1 deficient mice and small molecule antagonists indicate an important role of PAR1 in thrombosis and restenosis and thus the therapeutic potential of a PAR1 antagonist in treating these diseases. Development of a thrombin receptor tethered ligand analog binding assay led to the discovery of several different series of potent, nonpeptide small molecular antagonists of PAR1. These antagonists are PAR1 selective and inhibit most of the cellular effects of thrombin. A PAR1 antagonist has an advantage over a direct thrombin inhibitor since it does not inhibit enzymatic action of thrombin in the coagulation cascade with the consequent minimal bleeding side-effects, unlike a direct thrombin inhibitor. In addition, the emerging evidence for the role of PAR1 in various inflammatory diseases suggests as yet unexplored therapeutic potentials of PAR1 antagonists in various inflammatory diseases. PMID- 14529398 TI - Cyclooxygenase-2 biology. AB - In mammalian cells, eicosanoid biosynthesis is usually initiated by the activation of phospholipase A(2) and the release of arachidonic acid from membrane phospholipids in response to the interaction of a stimulus with a receptor on the cell surface. Arachidonic acid is subsequently transformed by the enzyme cyclooxygenase (COX) to prostaglandins (PGs) and thromboxane (TX). The COX pathway is of particular clinical relevance because it is the major target for non-steroidal anti-inflammatory drugs, which are commonly used for relieving inflammation, pain and fever. In 1991, it was disclosed that COX exists in two distinct isozymes (COX-1 and COX-2), one of which, COX-2, is primarily responsible for inflammation but apparently not for gastrointestinal integrity or platelet aggregation. For this reason, in recent years, novel compounds that are selective for this isozyme, the so-called selective COX-2 inhibitors or COXIBs, which retain anti-inflammatory activity but minimize the risk of gastrointestinal toxicity and bleeding, have been developed. This review article provides an overview and an update on the progress achieved in the area of COX-2 and PG biosynthesis and describes the role of COX-2 in health and disease. It also discusses some unresolved issues related to the use of selective COX-2 inhibitors as a safe and promising therapeutic option not only for the treatment of inflammatory states but also for cancer. PMID- 14529397 TI - Potential new targets for antithrombotic therapy. AB - Thrombosis is the collective term for diseases caused by the localized accumulation of circulating blood elements within the vasculature that result in vessel occlusion. Conventional antithrombotic drugs can inhibit thrombus growth by targeting coagulation pathways (e.g., heparin, warfarin) or platelet-dependent mechanisms (e.g., aspirin, clopidogrel). Thrombolytic agents (e.g., streptokinase) are used to degrade thrombi in situ, thereby restoring the blood flow. Despite advances, the search for new strategies continues because existing treatments impair hemostasis, and must be administered at dose levels that do not achieve maximum efficacy. Only a few drugs are used at markedly efficacious doses, for short periods of time in closely watched clinical situations, such as interventional cardiology and surgery. Ideally, new targets for therapy would lead to the development of agents that are specific for thrombus-forming mechanisms without affecting hemostasis. In the absence of such agents, new products should preferentially inhibit the thrombotic process at doses that are relatively safe. The symptomatology of naturally occurring or experimentally induced alterations of relevant hemostatic pathways can serve as basis for target selection. Hemostatic disorders that are compatible with life, do not pose a significantly increased risk of bleeding, but potentially protect against thrombosis provide guidance for rational design strategies. Theoretical considerations and recent experimental data suggest that: 1) inhibition of intrinsic coagulation pathway activity, 2) reduction of circulating platelet count, or 3) activation or enhancement of endogenous protein C or thrombolytic pathways could improve antithrombotic therapy. PMID- 14529399 TI - Dual COX inhibition and upper gastrointestinal damage. AB - Aspirin and non-aspirin NSAIDs injure the gastrointestinal tract principally as a result of their inhibition of prostaglandin synthesis. This is mediated via abrogation of the secretion of mucus and bicarbonate and by reduction in mucosal blood flow. Topical injury and inhibition of platelet thromboxane may also contribute respectively to damage and ulcer bleeding. Recognition of a second cyclooxygenase, COX-2, enabled drugs to be developed that selectively target this enzyme which is expressed in inflamed joints. These have proved to be effective treatments whilst causing little or no acute gastroduodenal injury and reduced ulcers and their complications. Future strategies may capitalise upon the phenomenon of substrate diversion of lipoxygenase products. Balanced cyclooxygenase/lipoxygenase inhibition maybe less harmful than cyclooxygenase inhibition. Also, nitric oxide can subserve many of the protective effects of prostaglandins and NO-donating NSAIDs are under evaluation. PMID- 14529400 TI - Gastroduodenal safety of cyclooxygenase-2 inhibitors. AB - Cyclooxygenase-1 (COX-1) derived eicosanoids promote gastroprotective mucosal defenses and induce platelet aggregation. By sparing COX-1, COX-2 specific inhibitors provide effective anti-inflammatory and analgesic activity while substantially reducing the risk of peptic ulcer disease and GI bleeding compared to dual COX inhibitors (traditional NSAIDs). Clinical studies of the COX-2 selective inhibitors have demonstrated efficacy equivalent to nonselective NSAIDs with significantly lower rates of GI toxicity. The incidence of endoscopic ulcers in some studies with coxibs has approximated placebo. However, as the detection of endoscopic lesions is not always correlated with symptomatic ulcers and ulcer complications, outcome studies of GI safety were performed. The results of large outcome studies have evaluated rofecoxib and celecoxib in over 39,000 patients with osteoarthritis or rheumatoid arthritis. Results of these studies showed that patients taking a supratherapeutic dose of rofecoxib or celecoxib had significantly lower rates of GI-related adverse events than those taking a nonselective NSAID. The GI safety of coxibs for patients using low dose aspirin concomitantly with a coxib appears to be reduced, particularly with regard to ulcer complications. Such data provide support for the COX-2 hypothesis and demonstrate that coxibs provide effective treatment of pain and inflammation with a reduced risk of gastropathy. PMID- 14529401 TI - Mucosal repair and COX-2 inhibition. AB - The healing of gastric ulcer is a complicated process that involves the proliferation of epithelial and endothelial cells and the concerted actions of a wide range of growth factors. Prostaglandins play an important role in ulcer healing. Expression of cyclooxygenase-2 (COX-2) is markedly upregulated around the margins of gastric ulcers and its inhibition leads to a delay of ulcer healing. Several of the growth factors that promote ulcer healing may work in part through COX-2-dependent mechanisms. Angiogenesis, which is crucial to ulcer healing, is tightly regulated by growth factors. Treatment with selective COX-2 inhibitors appears to alter the balance of serum levels of growth factors, favoring an inhibition of angiogenesis. Given the importance of COX-2 in regulating ulcer healing, caution should be taken in the use of selective inhibitors of COX-2 by patients at risk of ulcer disease. PMID- 14529402 TI - COX-2 inhibition, H. pylori infection and the risk of gastrointestinal complications. AB - Current data on the gastric safety of cyclooxygenase-2 (COX-2) inhibitors in the presence of H. pylori infection are largely derived from animal experiments and indirect clinical evidence. In animal models of H. pylori gastritis, COX-2 inhibitors suppressed prostaglandin synthesis and aggravated mucosal damage. In the human stomach, COX-1 appears to be the predominant source of prostaglandins despite the fact that COX-2 is upregulated in H. pylori gastritis. There are conflicting data on whether H. pylori alters the risk of ulcer in patients receiving COX-2 inhibitors. Among patients with H. pylori infection, rofecoxib reduced the risk of complicated gastric but not duodenal ulcers as compared to naproxen. The advantage of rofecoxib over naproxen also disappeared in patients with H. pylori infection and prior upper gastrointestinal events. In contrast, pooled data suggested that H. pylori increases the risk of ulcer in patients receiving nonselective nonsteroidal anti-inflammatory drugs but not in patients receiving celecoxib. In rodent gastric ulcers, COX-2 was upregulated in the granulation tissue and ulcer margin. Inhibition of COX-2 delayed healing of experimental gastric ulcer. Limited data showed that COX-2 expression was also increased in human gastric ulcer regardless of the H. pylori status. The functional significance of COX-2 in human gastric ulcer is unknown. PMID- 14529404 TI - COX-2 and colorectal cancer. AB - Metabolites of arachidonic acid participate in normal growth responses and in aberrant cellular growth and proliferation, including carcinogenesis. The key step in the conversion of free arachidonic acid to prostaglandins is catalyzed by the cyclooxygenase enzyme (COX). There are two COX enzymes, COX-1 and COX-2. COX 1 is expressed constitutively and is part of normal cell metabolic functions. COX 2, on the other hand, is induced and expressed in neoplastic growths. The connection between COX expression and carcinogenesis was first implicated in studies that demonstrated the efficacy of aspirin and non-steroidal anti inflammatory drugs to reduce the relative risk of colon cancer and also promote tumor regression in both humans and animal models of colon cancer. Investigation of the molecular basis of these observations showed that high levels of COX-2 protein were present in both human and animal colorectal tumors. A variety of evidence gathered from epidemiological, whole animal, and cellular studies indicate that unregulated COX-2 expression is a rate-limiting step in tumorigenesis and also that the loss of regulation occurs early in carcinogenesis. The interest in the COX-2 enzyme is that specific inhibition of COX-2 could theoretically avoid the gastrointestinal and other complications observed with the use of nonspecific COX inhibitors (most NSAIDs) or COX-1 inhibitors. The mechanisms by which COX-2 inhibitors lead to decreased colon carcinogenesis are not fully understood but they involve an increase not only in COX-2 dependent but also in COX-2 independent mechanisms. PMID- 14529403 TI - COX-2 inhibition versus gastroprotection with dual COX inhibitors: an evidence based approach. AB - Highly selective inhibitors of cyclooxygenase-2 (COX-2i) were introduced to minimize peptic ulcers and their complications caused by dual COX inhibitors (COXi). Co-prescribing a (generally cheap) dual COXi with a gastroprotectant is an alternative strategy, proven to reduce the incidence of NSAID-associated endoscopic ulcers. This review compares the efficacies of these two strategies and makes some estimates of their relative cost-effectiveness. In standard risk patients, endoscopic ulcers are reduced to about the same extent (around 70-80%) by either co-prescribing omeprazole or lansoprazole with a dual COXi or preferring a COX-2i alone. COX-2i reduced ulcer complications by a weighted mean of around 60% in comparative studies with dual COXi. There is little information about the influence of PPI on this endpoint, although one study using H. pylori treatment as a possible surrogate for placebo intervention found 77% protection against recurrent upper gastrointestinal bleeding by co-administered omeprazole. One direct comparison of the two strategies in high-risk patients (recent ulcer bleed) found quite high rates of re-presentation with bleeding ulcer using either strategy, and the differences between them were not significant. Drug costs in four Western countries were compared for each strategy. In one, the costs were similar, but in the others the combination of a cheap dual COXi with omeprazole was usually more expensive than using a COX-2i. The safest strategy in highest risk patients may be to co-prescribe a gastroprotectant with a COX-2i, with resulting higher drug costs but possibly offset by savings in other health costs. The efficacy and cost-benefit of this alternative approach warrants investigation. PMID- 14529405 TI - Clinical implications of COX-1 and/or COX-2 inhibition for the distal gastrointestinal tract. AB - Side effects of the distal gastrointestinal tract after NSAID use are common and more frequent than previously recognized. Increased mucosal permeability and mucosal inflammation are often silent but appear after NSAID treatment with most dual COX inhibitors. Other clinical manifestations include: anemia, occult blood loss, malabsorption, protein-loss, ileal dysfunction, diarrhea, mucosal ulceration and strictures due to diaphragm disease. More common complications are lower gastrointestinal bleeding and perforation, which represent at least one third of all gastrointestinal complications observed with NSAID use. Studies with selective COX-2 inhibitors have shown that, in the short term, these agents do not increase mucosal permeability or induce anemia due to occult bleeding and that, when compared to dual COX inhibitors, lower gastrointestinal complications may be reduced by 50%. In order to minimize the impact of these side effects, it is important to increase the current standards of suspicion by physicians who treat these patients, since drug discontinuation may further reduce damage, and clinical experience with agents that may prevent or treat distal tract damage is very limited. From this perspective, selective COX-2 inhibitors may be the drugs of choice in the high-risk patient that needs NSAIDs. Another important area of uncertainty is the impact of NSAID use in patients with inflammatory bowel diseases. Data from different animal models of inflammatory bowel disease suggest that inhibition of both COX-1 and COX-2 derived prostaglandins affects the severity of the mucosal inflammation. However, current epidemiological and clinical data are contradictory. Since many patients. PMID- 14529406 TI - Cyclooxygenase-2 inhibition and gastric cancer. AB - Epidemiological evidences suggest that chronic use of aspirin and nonsteroidal anti-inflammatory drugs (NSAIDs) might be associated with a reduced risk of gastrointestinal cancers, including gastric cancer. The pre-cancerous gastric lesions and gastric cancers over-expressed cyclooxygenase (COX)-2. This overexpression not only is associated with Helicobacter pylori infection, but also maybe due to exposure to carcinogens. Targeted inhibition of COX, especially the COX-2 isoform, can lead to growth inhibition and apoptosis of gastric cancer in vitro. Various mechanisms, including COX-dependent and COX-independent pathways, have been identified and will be discussed in this article. Animal xenograft models have confirmed the tumor suppressing effects of COX-2 inhibitors. Human studies are underway to examine the use of COX-2 inhibitor in the treatment of pre-cancerous lesions. COX-2 inhibitors have a promising role in the prevention and treatment of gastric cancer. PMID- 14529407 TI - CB 1954: from the Walker tumor to NQO2 and VDEPT. AB - CB 1954 [5-(aziridin-1-yl)-2,4-dinitrobenzamide] has been the subject of continued interest for over 30 years. As an anti-cancer agent, it represents one of the very few examples of a compound that shows real anti-tumor selectivity. Unfortunately, for the treatment of human disease, this anti-tumor selectivity was seen only in certain rat tumors. The basis for the anti-tumor selectivity of CB 1954 is that it is a prodrug that is enzymatically activated to generate a difunctional agent, which can form DNA-DNA interstrand crosslinks. The bioactivation of CB 1954 in rat cells involves the aerobic reduction of its 4 nitro group to a 4-hydroxylamine by the enzyme NQO1 (DT-diaphorase). The human form of NQO1 metabolizes CB 1954 much less efficiently than rat NQO1. Thus human tumors are insensitive to CB 1954. In view of the proven success of CB 1954 in the rat system, it would be highly desirable to re-create its anti-tumor activity in man. This has led to the development of CB 1954 analogs and other prodrugs activated by nitroreduction such, as those based on a self-immolative activation mechanism. A gene therapy-based approach for targeting cancer cells and making them sensitive to CB 1954 and related compounds has been developed. VDEPT (gene directed enzyme prodrug therapy) has been used to express an E. coli nitroreductase in tumor cells and human tumor cells transduced to express this enzyme are very sensitive to prodrugs activated by nitroreduction. CB 1954 is in clinical trial for this application. Recently it has been shown that a latent nitroreductase is present in some human tumors. This is NQO2--an enzyme that requires for activity, the non-biogenic compound dihydronicotinamide riboside (NRH) as a cosubstrate. When active, NQO2 is 3000 times more effective than human DT-diaphorase in the reduction of CB 1954. NRH and reduced pyridinium derivatives that, like NRH, act as co-substrates for NQO2, produce a dramatic increase in the cytotoxicity of CB 1954 against human cell lines in vitro and its anti-tumor activity against certain human xenografts in vivo. NQO2 activity is substantially raised in tumor samples from colorectal and hepatoma patients (up to 14-fold). A phase I clinical trial of an NQO2 co-substrate with CB 1954 is scheduled. PMID- 14529408 TI - Radiogenetic therapy: strategies to overcome tumor resistance. AB - The aim of cancer gene therapy is to selectively kill malignant cells at the tumor site, by exploiting traits specific to cancer cells and/or solid tumors. Strategies that take advantage of biological features common to different tumor types are particularly promising, since they have wide clinical applicability. Much attention has focused on genetic methods that complement radiotherapy, the principal treatment modality, or that exploit hypoxia, the most ubiquitous characteristic of most solid cancers. The goal of this review is to highlight two promising gene therapy methods developed specifically to target the tumor volume that can be readily used in combination with radiotherapy. The first approach uses radiation-responsive gene promoters to control the selective expression of a suicide gene (e.g., herpes simplex virus thymidine kinase) to irradiated tissue only, leading to targeted cell killing in the presence of a prodrug (e.g., ganciclovir). The second method utilizes oxygen-dependent promoters to produce selective therapeutic gene expression and prodrug activation in hypoxic cells, which are refractive to conventional radiotherapy. Further refining of tumor targeting can be achieved by combining radiation and hypoxia responsive elements in chimeric promoters activated by either and dual stimuli. The in vitro and in vivo studies described in this review suggest that the combination of gene therapy and radiotherapy protocols has potential for use in cancer care, particularly in cases currently refractory to treatment as a result of inherent or hypoxia-mediated radioresistance. PMID- 14529409 TI - Enzyme/abzyme prodrug activation systems: potential use in clinical oncology. AB - Clinically useful prodrug activation systems for cancer therapy can be applied in combination with the exogenous activating enzymes, by which masked prodrugs are able to unmask to exert cytotoxic effects on the target tumors. In essence, designing prodrugs not to be degenerated or activated by the endogenous enzymes is needed. Prodrug activation systems are to be delivered to the tumor site by delivery tools, including antibodies, genes, viral vectors and synthetic polymers, directed to the target tumors. Highly selective accumulation of the prodrug activation system at the tumor site is critically important for the efficacy of the prodrug activations. Genetic engineering of antibodies have made it possible to create a bispecific antibody and its derivatives, which are of special value to the functional antibodies with one arm to direct the target tumor tissues, and another to recruit the effector cells or molecules that can effectively kill the tumor cells. The technology has further opened the window for catalytic antibodies as a prodrug activating system. Catalytic antibodies have two distinct advantages over the enzymes: First, they can be selected to catalyze the reaction that is not catalyzed by the endogenous enzymes. Second, in order to minimize immunogenicity, humanization is applicable to catalytic antibodies. In viewing the concept and experimental data with a few clinical trials of recent approaches of prodrug activation systems, their potential utility in clinical oncology is further discussed. PMID- 14529411 TI - Enzyme and proton-activated prodrugs for a selective cancer therapy. AB - This review is a survey of two approaches for a selective anticancer therapy that are based on a specific cleavage of specially designed non-toxic prodrugs with the liberation of a cytotoxic compound either by antibody-enzyme conjugates targeted to tumor-associated antigens or by acid-catalyzed hydrolysis of the prodrugs due to the increased concentration of hydronium ions in malignant tissue under hyperglycemic conditions. Herein, the design, synthesis and the biological testing of prodrugs are described. PMID- 14529410 TI - Prodrugs in genetic chemoradiotherapy. AB - Improvements in the radiotherapeutic management of solid tumors through the concurrent use of gene therapy is a realistic possibility. Of the broad array of candidate genes that have been evaluated, those encoding prodrug-activating enzymes are particularly appealing since they directly complement ongoing clinical chemoradiation regimes. Gene-Directed Enzyme-Prodrug Therapy (GDEPT) only requires a fraction of the target cells to be genetically modified, providing that the resultant cytotoxic prodrug metabolites redistribute efficiently (the bystander effect). This transfer of cytotoxicity to neighboring non-targeted cancer cells is central to the success of any gene therapy strategy, irrespective of the therapeutic gene employed. In the context of genetic chemoradiotherapy, efficient prodrug metabolite diffusion will be a prerequisite for efficient radiosensitization. Some, but not all GDEPT approaches have been analysed in combination with radiotherapy. Examples of prodrugs of clinically established chemotherapeutic agents currently used in conjunction with radiotherapy include: 5-fluorocytosine (5FC), cyclophosphamide (CPA), irinotecan (CPT-11), gemcitabine (dFdC), capecitabine, mitomycin C (MMC) and AQ4N. Other GDEPT paradigms, such as ganciclovir (GCV) and Herpes Simplex thymidine kinase (HSV-tk), dinitrobenzamide (DNB) mustard or aziridinyl analogs and the E. coli nitroreductase (NTR), CMDA or ZP2767P with Pseudomonas aeruginosa carboxypeptidase G2 (CPG2), and indole-3-acetic acid (IAA) activated by horseradish peroxidase (HRP) have no clinically established chemotherapeutic counterpart. Each prodrug is discussed in this review in the context of GDEPT, with a particular attention to translational research and clinical utility in combination with radiotherapy. PMID- 14529412 TI - Identification and targeting of tumor escape mechanisms: a new hope for cancer therapy? AB - Conventional cancer therapy is administered in the form of surgery, radiotherapy or chemotherapy. Immunotherapy is the latest asset to the panel of anti-cancer treatments. This approach appears favorable over the other more conventional methods for various reasons: (1) it is highly specific for cancer cells and, therefore, low toxicity should be expected; (2) it recognizes and eliminates cancer cells regardless of their phase in the cell cycle; (3) tumors that developed drug resistances would still be a suitable target for immunotherapy. (4) Immunotherapy offers the possibility of preventive immunization of high-risk patients. Due to the diverse mechanisms that result in the transformation of cells and subsequent tumor development, not all cancers respond similarly to treatment. Significant effort is currently invested in the characterization of the underlying regulatory network in individual cancers responsible for tumorigenesis. Understanding tumors better allows on one hand the identification of essential pathways that can be intercepted to kill the transformed cells more specifically. On the other hand, these insights also allow us to exclude therapeutic strategies with little chance of success when dealing with tumor escape mutants thus saving valuable time and resources. Any tumor therapy puts selective pressure on tumors thus favoring the outgrowth of therapy-resistant variants. This review summarizes current knowledge on tumor escape mechanisms and some of the efforts to overcome these mechanisms. PMID- 14529413 TI - Porphyrins as radiosensitizing agents for solid neoplasms. AB - The biological effects of radiation affect both neoplastic and normal tissues. The nature and extent of such effects, however, depend on selected biological parameters (e.g., oxygen supply, cell cycle) and can be modified by chemical agents such as radiosensitizers, radioprotectors and chemotherapeutic agents. A precise control of the mode of action of the radiation is important in order to achieve the maximum effect on tumor tissue, while minimizing the effect on normal tissues. Most of the known and routinely used radiosensitizers are neither selective nor tumor specific. This article reviews a new selective and specific modality that increases the sensitivity of solid tumor tissue, especially of radio resistant, hypoxic tumor cells, to radiation. This modality is currently under early clinical evaluation and encompasses the application of Photofrin II, which is already used as a photosensitizer in photodynamic therapy (PDT) at predetermined times prior to irradiation. PMID- 14529414 TI - Bradykinin antagonists as anti-cancer agents. AB - The nonapeptide bradykinin is an important growth factor for many cancers. Certain peptide and non-peptide bradykinin antagonists show remarkable anti cancer activities in both in vitro and in vivo cancer models, especially of lung and prostate cancers. Bradykinin antagonists stimulate apoptosis in cancers by a novel "biased agonist" mechanism: they block intracellular increase of calcium concentration but stimulate the MAP kinase pathway. This unbalanced effect stimulates caspase activation. In nude mouse xenotransplants of lung and prostate cancers the antagonists inhibit angiogenesis and activation of membrane metalloproteases (MMP 2 and 9). In the xenotransplants certain bradykinin antagonists showed higher potency than standard anti-cancer drugs, without evident toxicity to the hosts. These compounds offer great promise for development of new anti-cancer drugs. PMID- 14529415 TI - SRC family kinases: potential targets for the treatment of human cancer and leukemia. AB - The inherited or acquired deregulation of protein kinase activity has been implicated in the pathogenesis of many human diseases, including cancer. Therefore, the inhibition of kinases has been proposed to be a promising strategy in the context of anti-cancer treatment. Many other kinases have been selected as drug discovery targets based on the prevalence of mutations, over-expression and unscheduled activation in human cancer. Of the various protein kinases chosen, Src family kinases are amongst the most extensively studied kinase oncogenes in academia and industry. This review focuses on our current understanding of the deregulation and role of Src family kinases in human cancer and leukemia. Recent data implicate the action of c-Src in cancer metastasis, mediated by up regulation of various protease systems (calpain, uPA) as well as disruption of E cadherin signalling. Moreover, novel roles of various Src family members in the development of human leukemia have been found. New insights into downstream signalling mechanisms, including the activation of STAT3, PDK1 and Akt, further corroborate the importance of Src family kinases in tumorigenesis and chemoresistance. Despite our rather clear understanding of Src family kinases as pro-oncogenes no Src family kinase inhibitor has entered a clinical trial so far. This review will discuss prerequisites to be fulfilled for clinically targeting c Src and its homologues using small molecule drugs. PMID- 14529416 TI - Renaissance of the biologically active vitamin A derivatives: established and novel directed therapies for cancer and chemoprevention. AB - Vitamin A and its biologically active derivatives are involved in a complex arrangements of physiological and developmental responses in many tissue of higher vertebrates. Retinoids are natural and synthetic compounds related to retinoic acid that act through interaction with two basic types of nuclear receptors: retinoic acid receptors (RAR alpha, RARbeta and RARgamma) and retinoid X receptors (RXRalpha, RXRbeta and RXRgamma) as retinoid-inducible transcription factors. Thus, the retinoid receptors are considered to be ligand-activated, DNA binding, trans-acting, transcription-modulating proteins involved in a general molecular mechanism responsible for transcriptional responses in target genes. They exert both beneficial and detrimental activity; they have tumor-suppressive activity but on the other hand they are teratogenic. Retinoids inhibit carcinogenesis, suppress premalignant epithelial lesions and tumor growth and invasion in a variety of tissues. Natural and synthetic retinoids have therapeutical effects due to their antiproliferative and apoptosis-inducing effects. They are known to cause redifferentiation or to prevent further dedifferentiation of various neoplastic tissues. A number of novel chemical compounds, receptor selective retinoids and rexinoids, have been synthesized up to now and tested both in vitro and in vivo, using animal models against different cancer cells. In spite of that progress, there is still an urgent call for novel synthetic retinoids and rexinoids with greater retinoid receptor selectivity, reasonable chemotherapeutic or chemopreventive effects and reduced toxicity and side effects. This article summarizes selected effects of biologically active natural or synthetic retinoids and rexinoids, acting through their cognate nuclear receptors, and their use in chemotherapy and chemoprevention of various types of cancer. PMID- 14529417 TI - Recent developments in the field of tumor-inhibiting metal complexes. AB - 25 years after the first approval of cisplatin in the clinic against a number of cancer diseases, cisplatin and related compounds continue to be among the most efficient anticancer drugs used so far. Efforts are focused to develop novel platinum- and non-platinum-based antitumor drugs to improve clinical effectiveness, to reduce general toxicity and to broaden the spectrum of activity. In the field of non-platinum compounds exhibiting anticancer properties, ruthenium complexes are very promising, showing activity on tumors which developed resistance to cisplatin or in which cisplatin is inactive. Furthermore, general toxicity was found to be very low. The first ruthenium compound NAMI-A entered phase I clinical trials in 1999 as an antimetastatic drug, whereas the ruthenium complex KP1019 will enter phase I clinical trials in 2003 as an anticancer drug which is among others very active against colon carcinomas and their metastases. Remarkable progress is also seen in developing tumor inhibiting gallium compounds. One of them, KP46, will also enter phase I clinical trials in 2003. This article reviews briefly the achievements in the field of anticancer metal complexes focusing the discussion onto the impact of the group of Bioinorganic Chemistry at the Department of Inorganic Chemistry at the University of Vienna. The development of pH sensitive platinum prodrugs, platinum-based drug targeting strategies with low-molecular-weight carriers, kinetically inert platinum(IV) complexes, as well as tumor inhibiting non platinum anticancer drugs based on ruthenium and gallium is covered in the following sections. PMID- 14529418 TI - Safety and utilization of blood components as therapeutic delivery systems. AB - In recent years, natural blood components have been extensively studied as the advanced therapeutic delivery systems. The blood components which can potentially be used as the therapeutic delivery systems include different types of cells, such as erythrocytes and lymphocytes, macromolecular complexes such as lipoproteins and antibody or albumin conjugates and other molecules. This review article covers the progress in this topic, specifically, including the safety issues and the utilization of these component. It can be seen through the literature that the blood components as the therapeutic delivery systems have a number of advantages over traditional pharmaceutical products. The efficacy and practice of the applications, however, require significant amount of development work in the near future. PMID- 14529419 TI - Recent trends in the use of polysaccharides for improved delivery of therapeutic agents: pharmacokinetic and pharmacodynamic perspectives. AB - New and innovative methods of delivery of therapeutic agents using polysaccharides have been recently developed, which target site of action, increase the intensity and/or prolong pharmacologic action, and/or reduce toxicity of small molecule drugs, proteins, or enzymes. This review is focused on the role of dextran, pullulan, and mannan polysaccharides in such applications. While dextran and pullulan are glucose polymers with different glucosidic linkages, mannan is composed of mannose units. In terms of pharmacokinetics of the carriers themselves, molecular weight (MW), electric charge, various chemical modifications, and degree of polydispersity and/or branching would mostly determine their fate in vivo. Generally, large MW polysaccharides (MWs > or = 40 kD) have low clearance and relatively long plasma half life, resulting in accumulation in reticuloendothelial or tumor tissues. The tumor accumulation in most cases is a passive targeting due to "enhanced permeation and retention" of macromolecules by tumors. Additionally, drugs such as anticancer agents may be actively targeted to specific cells by polysaccharides to which appropriate ligands are attached. In terms of mode of use, polysaccharides have been utilized in a variety of innovative ways for improvement of drug delivery. Their most important application has been as carriers for preparation of macromolecular prodrugs that are normally inactive and need to release the active drug at the site(s) of interest. Also, they have been used for preparation of macromolecule protein conjugates, which may retain the activity of the proteins, in order to increase the duration of effect and decrease the immunogenicity of proteins. Several other new applications, such as polysaccharide-anchored liposomal formulations, have also been gained attention recently and are briefly reviewed here. Finally, four recent examples of polysaccharide-based delivery systems involving specific drugs/imaging agents are reviewed in detail in terms of their development, pharmacokinetics, and pharmacodynamics. Collectively, these data suggest that macromolecular polysaccharides are promising agents for improving drug delivery. PMID- 14529420 TI - Application of chitin and chitosan derivatives in the pharmaceutical field. AB - Chitin and chitosan derivatives are used as excipients and drug carriers in the pharmaceutical field. Their derivatization contributed to expansion of application and decrease toxicity. Chitosan is used as an excipient in oral dosage form. Chitosan tablet can exhibit a sustained drug release compared to commercial products. Films prepared using chitin or chitosan have been developed as wound dressings, oral mucoadhesive and water-resisting adhesive by virtue of their release characteristics and adhesion. Intratumoral administration of gadopentetic acid-chitosan complex nanoparticles (approximately 430 nm in diameter) has been more effective for gadolinium neutron-capture therapy compared with a group treated with the solution. Compared to intragastrical feeding with diphtheria toxoid (DT) in PBS, a strong enhancement of the systemic (IgG) and local (IgA) immune responses against DT has been observed in mice fed with DT loaded chitosan microparticles (approximately 4.7 microm in size). When DNA loaded chitosan microspheres (1.15 - 1.28 microm) were intramuscularly administrated into mice, high beta-galactosidase and luciferase productions were obtained even after a long post-transfection period (12 weeks). N-Succinyl chitosan (Suc-Chi) has been studied for cancer chemotherapy as a drug carrier and the conjugates of mitomycin C with Suc-Chi exhibited good antitumor activities against various tumors. Furthermore, trimethyl-chitosan and monocarboxymethyl chitosan has been shown to be effective as intestinal absorption enhancers due to their physiological properties. Chitosan-thioglycolic acid conjugates has been found to be a promising candidate as scaffold material in tissue engineering due to their physicochemical properties. This review summarizes the application of chitin and chitosan derivatives for hospital preparations and drug carriers. PMID- 14529421 TI - Clinical implications of N-(2-hydroxypropyl)methacrylamide copolymers. AB - Different anticancer drugs, farmorubicin, doxorubicin, paclitaxel and cis-platin have been conjugated through a Gly-Phe-Leu-Gly tetrapeptide side chain to a water soluble synthetic polymeric carrier based on N-(2-hydroxypropyl)methacryalmide (HPMA) non-targeted or targeted with galactosamine and/or human IVIg and used in Phase I clinical trials. Conjugation of the drugs to the polymeric carrier that is non-toxic and non-immunogenic in man significantly decreased their non specific organ toxicities and increased maximum tolerated dose up to 5 times. Macromolecular therapeutics based on HPMA have radically different pharmacokinetics. Drugs are not released from their polymeric carrier and remain in the peripheral blood and urine of patients mostly in their polymer-bound form. A clinical response against some refractory cancers was recorded in Phase I clinical trials. It was also demonstrated that doxorubicin-HPMA copolymer conjugates containing an immunoglobulin moiety have both cytostatic and immunomobilizing activity. PMID- 14529422 TI - Contribution of poly(amino acids) to advances in pharmaceutical biotechnology. AB - Recently, protein biotechnology generates tremendous impacts in therapeutic products. These products include enzymes, antibodies, hormones, blood factors, growth factors and regulatory factors. Protein, vaccine and gene therapy drugs could be formulated with suitable biomaterials to deliver active agents to their target sites at the right time and maintain therapeutic effects for proper durations. In this review article, we focus on poly(amino acids) or polymerized amino acids for their applications in drug delivery systems, vaccines, and gene therapy. The nomenclatures of poly(amino acids) are briefly introduced to systematically express synthetic polypeptides. In drug delivery systems, we introduce two applications of poly(amino acids) in pharmaceutical biotechnology, either as carriers to facilitate drug delivery, or as biomaterials to be formulated as suitable delivery systems for application in tissue engineering. Many short polypeptides are mapped from antigen motifs and used for vaccination. These poly(amino acids) provide protective effects in animal challenge tests and potential application in vaccine development to be briefly introduced. Finally, some reports related to new developed poly(amino acids) as DNA carriers for achieving gene delivery are also described in the text. PMID- 14529423 TI - Non-degradable biocompatible polymers in medicine: past, present and future. AB - Polymers have a long history in medicine. Their uses to date range from traditional applications such as catheters, syringes, blood contacting extra corporeal devices to matrices for drug delivery, cell encapsulation and tissue regeneration. Polymers can be broadly classified on the basis of the reactivity of their chemical backbone (or susceptibility of the backbone to breakdown upon exposure to water, i.e., hydrolysis) as non-degradable and degradable. In this review, the polymers that exhibit no to very low degradation in aqueous and biological environments will be covered. The applications of various polymers both in traditional and emerging medical areas is discussed in the context of its chemical structure to better enable material selection for biomedical research. PMID- 14529424 TI - Intelligent polyelectrolytes and gels in oral drug delivery. AB - The present review concerns smart, or intelligent polymers for oral administration that change conformation in aqueous solutions in response to external stimuli such as pH or temperature. We concentrate on charged polymers and gels with polyelectrolyte properties. Because of the ionization at a certain pH or in response to changes in the ionic composition of the solution, a polyelectrolyte has better chances of displaying smart properties than a neutral polymer. When such smart polyelectrolyte is cross-linked by covalent or hydrogen bonding and/or physical aggregation or is entangled, it forms an environmentally sensitive gel capable of swelling and collapse in an aqueous medium. Varying pH, temperature, and microbial flora are found in the gastrointestinal tract, and thus pH- sensitive polymers and gels that can be degraded by specific enzymes and/or inhibit proteolytic enzymes can be tailored for the efficient site specific therapy. Smart polymers wield a lot of promise in the targeted, site specific administration where they can provide advantages in loading of sensitive drugs such as proteins and peptides, while releasing the drug at a specific pH or in response to the presence of certain microbial flora. PMID- 14529425 TI - Rituxan immunotherapy and zevalin radioimmunotherapy in the treatment of non Hodgkin's lymphoma. AB - Immunotherapy with the anti-CD20 monoclonal antibody rituximab has been shown in clinical trials to be effective in the treatment of both indolent and aggressive non-Hodgkin's lymphomas (NHL). Recent studies have demonstrated improved clinical benefit with extended dose and maintenance therapies in patients with indolent lymphomas and chronic lymphocytic leukemia. Rituximab's label was recently expanded to include treatment of bulky disease, retreatment of patients previously treated with rituximab, and an eight-week extended treatment schedule. Rituximab has also been effectively combined with chemotherapy, resulting in higher response rates and longer response durations in randomized trials in patients with aggressive lymphoma. Studies continue to evaluate and expand the role of rituximab in the treatment of NHL, including its use in combined immunotherapy approaches and autologous stem cell transplant as well as in the treatment of autoimmune disorders. Radioimmunotherapy with the rituximab and ibritumomab tiuxetan (Zevalin) regimen was recently approved for the treatment of relapsed or refractory low-grade, follicular or CD20+ transformed NHL, including rituximab refractory follicular NHL. The regimen is delivered on an outpatient basis over the course of a week. Studies are currently exploring sequential dose therapy, radioimmunotherapy with rituximab maintenance, and ibritumomab tiuxetan radioimmunotherapy as part of autologous stem cell transplant. Current understanding of the mechanisms of action of rituximab and the use of rituximab and ibritumomab tiuxetan in patients with indolent and aggressive NHL will be discussed. PMID- 14529426 TI - Intravenous immunoglobulin preparations and autoimmune disorders: mechanisms of action. AB - Intravenous immunoglobulins (IVIg) have been used as a substitutive treatment for primary and secondary humoral immune deficiencies for several decades. In the meantime, increased experience has been acquired with IVIg in the management of other inflammatory and autoimmune disorders, such as Kawasaki's disease, idiopathic thrombocytopenic purpura, dermatomyositis or Guillain-Barre syndrome, in which several clinical trials have demonstrated its efficacy. In other pathologies, IVIg seem to be effective, although further studies are required. Nevertheless, the exact mechanism by which IVIg exert their beneficial actions is not completely understood. According to in vitro as well as in vivo data, several mechanisms of action have been proposed: Fc receptor blockade, idiotype-anti idiotype interactions, neutralisation of bacterial toxins and superantigens, competitive inhibition of complement activation, down-regulation of B- and T-cell function, enhancement of pathogenic autoantibodies clearance, modulation of soluble products, apoptosis blockade via Fas receptor and administration of soluble products which could interfere with the immune response. Both IVIg structure, as well as its obtention from pooled human plasma donors, seem to play an important role in IVIg immunomodulatory properties. Thus, the objective of the present article is to review the current evidence upon the mechanisms of action of IVIg. PMID- 14529427 TI - The methylerythritol phosphate pathway and its significance as a novel drug target. AB - Isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) are the precursors for all isoprenoid compounds. Two pathways are found in Nature for their biosynthesis. The mevalonate (MVA) pathway is found in eukaryotes, algae, archae and some gram-positive bacteria. Gram-negative bacteria, plants and some gram-positive bacteria utilize the methyl erythritol phosphate (MEP) pathway. The distribution and the orthogonal nature of the pathways make the MEP pathway an attractive new target for antibiotics and herbicides. The MEP pathway is essential for bacterial viability. Inhibitors to the MEP pathway represent a "dual-use technology" because potential targets include potential biological warfare agents in addition to common human pathogens. The CDC has three categories designated for Biological Diseases/Agents. Three of the six entities designated as the highest priority (Category A) are organisms that utilize, or appear to utilize, the MEP pathway. Among the 12 second highest priority agents (Category B) listed, 8 are organisms that appear to utilize the MEP pathway. Common human pathogens that can be targeted include the organisms responsible for peptic ulcers, tuberculosis, malaria, food safety threats, and sexually transmitted diseases. There is so far only one inhibitor reported that specifically blocks the MEP pathway and is being investigated clinically. This compound, fosmidomycin, has been shown to be somewhat effective in treating Plasmodium falciparum, the parasite responsible for malaria. We foresee that new MEP pathway inhibitors will open up an entirely new class of antibiotics. An MEP pathway intermediate has also been shown to be the most potent gammadelta T cell activator. PMID- 14529428 TI - Roles of growth factors in chemotherapy-induced intestinal mucosal damage repair. AB - Chemotherapy agents induce apoptotic cell death and loss of cell proliferation in the intestinal crypt epithelium, resulting in intestinal mucosal damage called "mucositis". Small intestinal mucositis is characterized structurally by crypt loss and villus atrophy, and functionally by absorptive and barrier impairments. The increased use of chemotherapy in cancer treatment and the clinical importance of the intestinal mucositis as a common side effect have stimulated more active research into understanding the pathophysiology of intestinal mucositis and developing agents for preventing or treating this condition. Rodent studies have shown that, following the chemotherapy-induced initial apoptosis and loss of crypt cell proliferation, many different growth factors or their receptors are upregulated locally at the crypts, preceding or coinciding with the epithelial hyperproliferative repair response. Aiming to reduce crypt cell apoptotic sensitivity to cytotoxic chemotherapy and/or to enhance crypt epithelial proliferative repair, several exogenous growth factor treatments have been tested, either preclinically and/or clinically, and are showing promise for their efficacy or safety in preventing or treating chemotherapy-induced mucositis. These tested growth factors include keratinocyte growth factor, interleukin-11, transforming growth factor beta, milk-derived growth factor extract, macrophage/granulocyte colony stimulating factors, and glucagon-like peptide 2. Further research on the basic and discovery levels and subsequent translational studies are needed to understand more about chemotherapy-induced intestinal mucositis and to identify candidates of growth factors or other agents that will potentially prevent or treat chemotherapy-induced mucositis more effectively, specifically, safely, and practically in chemotherapy patients. PMID- 14529429 TI - Peripheral opioid analgesia. AB - Opioids have long been thought to act exclusively within the central nervous system. An increasing number of studies recently reported the existence of opioid receptors outside the central nervous system and therefore suggested that opioids are also able to produce analgesic effects in the periphery. Such effects are particularly prominent under painful inflammatory conditions, both in animals and in humans. During inflammatory processes, opioid receptors are transported from dorsal root ganglia towards the peripheral sensory nerve endings. At the same time, immune cells containing endogenous opioid peptides accumulate within the inflamed tissue. Environmental stimuli (e.g. stress) as well as releasing agents (e.g. corticotropin releasing factor, cytokines) can liberate these opioid peptides to interact with the neuronal opioid receptors and elicit local analgesia. The inflammation-induced activation of opioid production and the release of endogenous opioids from immune cells may lead to novel approaches for the development of peripherally acting analgesics. Clinical investigation now focuses on the development of new peripheral opioid agonists as well as on ways to stimulate the endogenous analgesic system in order to induce effective peripheral analgesia with reduced central side effects typically associated with opioids. PMID- 14529446 TI - Carboranylporphyrins for boron neutron capture therapy of cancer. AB - A major challenge for cancer treatment is the preferential and irreversible killing of tumor cells and minimal damage of normal tissues, both in the site of the malignancy and in the body. The agents used in boron neutron capture therapy (BNCT) are supposed to have the following advantages over many conventional chemotherapeutics: 1) when irradiated with thermal neutrons, an unstable isotope (11)B is formed whose rapid decay yields local and a thermal effect; 2) because the free path of the released particles is close to the cell diameter, the tissues outside the tumor should gain less damage; 3) local radioactivity and heat should be harmful for cells that, in the course of their natural history, acquired the determinants of altered response to many toxic stimuli. However, a higher specificity of damage would be achieved if the drugs accumulate mostly in cancer cells rather than in non-malignant counterparts. Therefore, optimization of agents for BNCT presumes the design of chemicals with improved accumulation/ retention in cancer cells. In particular, carboranyl-substituted porphyrins, the stable conjugates of macrocyclic porphyrins with complex boron-containing polyhedra, are considered good candidates for BNCT due to their uptake by cancer cells and high boron content. Importantly, the proposed mechanisms of pharmacological effects of carboranylporphyrins make these compounds potentially appropriate for elimination of pleiotropically resistant tumor cells. PMID- 14529447 TI - Current and future directions in the treatment of metastatic malignant melanoma. AB - Recently treatment strategies in advanced malignant melanoma have significantly changed. Due to high response rates (e.g. more than 50% for the Dartmouth regimen), combination chemotherapy has been the standard therapy in several oncological and dermatooncological centers in the USA and Europe. For the last three years different prospective randomized phase III trials failed to achieve similar results. There was no benefit in overall survival and in response duration in comparison to single agent chemotherapy. Currently, randomized clinical trials seem to be the best approach for the clinical treatment of metastatic melanoma. In this review several novel strategies against malignant melanoma are discussed with focus on the role of single agent chemotherapy and biochemotherapy. PMID- 14529448 TI - GnRH agonists and antagonists in cancer therapy. AB - Gonadotropin releasing hormone (GnRH) is a hypothalamic decapeptide that binds to GnRH receptors on pituitary gonadotrope cells to modulate the synthesis and secretion of the gonadotropins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH). These gonadotropins in turn regulate gonadal steroidogenesis and gametogenesis. Chemical characterization and structure-activity analysis of GnRH variants containing systematic amino acid substitutions led to the discovery of GnRH superagonists and antagonists. These peptides are widely used for the treatment of clinical conditions in which modulation of or interference with sex hormone production is beneficial to prevent development or progression of benign conditions (e.g. endometriosis, uterine fibroids) or malignant tumors (e.g. breast, ovarian, endometrial and prostate carcinoma). When compared to native GnRH, GnRH superagonists have increased potency for the short-term release of gonadotropins. However, they show paradoxical action in that chronic treatment with superagonists results in inhibition of gonadotropin production as a result of desensitization of the gonadotropes and down regulation of its receptor. In contrast, GnRH antagonists produce a rapid and dose-dependent suppression of gonadotropin release by competitive blockade of the GnRH receptors without any initial stimulatory effect as seen with superagonists. In recent years, a search for peptidomimetic compounds to replace peptides as therapeutic agents has been undertaken to find compounds with higher affinity for the GnRH receptor but do not have the disadvantages of peptides. Such efforts have resulted in the identification and development of small-molecule non-peptide compounds that are sufficiently stable in vivo and possess favorable pharmacological parameters comparable to peptide antagonists. Some of these compounds are being tested in human volunteers and the preliminary results are very encouraging. PMID- 14529449 TI - Chartreusin, elsamicin A and related anti-cancer antibiotics. AB - Chartreusin and elsamicin A are structurally related antibiotics that bind to GC rich tracts in DNA, with a clear preference for B-DNA over Z-DNA. They inhibit RNA synthesis and cause single-strand scission of DNA via the formation of free radicals. Elsamicin A can also be regarded as the most potent inhibitor of topoisomerase II reported so far. It can inhibit the formation of several DNA protein complexes. Elsamicin A binding to the P1 and P2 promoter regions of the c myc oncogene inhibits the binding of the Sp1 transcription factor, thus inhibiting transcription. Despite the pharmacological interest in chartreusin, elsamicin A and their derivatives, there is no experimental data on the structure of their complexes with DNA. This shortcoming has been partially solved by a theoretical approach, which provided some details about the DNA-elsamicin A interaction, and the thermodynamic characterization of the binding of chartreusin and elsamicin A to DNA. Elsamicin A but not chartreusin is being developed clinically as an anti-cancer agent. IST-622 (6-O-(3-ethoxypropylonyl)-3',4'-O-exo benzylidene-chartreusin), a novel semi-synthetic derivative of chartreusin, which has shown a promising anti-cancer activity in a phase II study, appears to be a pro-drug with a more suitable pharmacokinetic profile than chartreusin. PMID- 14529450 TI - Topoisomerases and tubulin inhibitors: a promising combination for cancer treatment. AB - Modern approaches to treatment of cancer seek to activate the internal suicide program in the malignant cells, and thereby effectively eliminate them without engaging most of other bodily systems. Many currently used cytostatics are known to induce apoptosis and efforts are being paid to develop new ones with better and more effective proapoptotic potential. Nevertheless, despite recent developments in this field, there are still numerous malignancies showing a varying degree of resistance to cell death due to the corrupted signaling pathways and genetic alterations, often in conjunction with expansive proliferation rate. It has been shown that topoisomerase inhibiting agents such as etoposide, camptothecin and others represent a powerful and dynamic group of cytostatic chemicals used in experimental and clinical conditions. So, it is a group of microtubule targeting poisons comprising classical colchicines on the one hand and new taxanes on the other hand. Since several members of both groups have been evidenced as apoptosis inducers operating via distinct mechanism, their combination should theoretically enhance the final therapeutic outcome. This minireview focuses on the possibilities of such a combinational approach with respect to possible benefits and hazards of this strategy. PMID- 14529451 TI - 4-oxa-1-azabicyclo[3.2.0]heptan-7-one derivatives as anti-tumor agents. AB - A series of naturally occurring and synthetic novel oxapenam (4-oxa-1 azabicyclo[3.2.0] heptan-7-one) derivatives with their antitumor activity and the structure-activity relationship among this class of compounds is reported. Among the synthetic 4-oxa-1-azabicyclo[3.2.0]heptan-7-one having an ester, amide, ether derivatives of hydroxy group at C-3 position exhibited either no activity or reduced the antitumor activity in vitro. The 3-amino acid 4-oxa-1 azabicyclo[3.2.0]heptan-7-one derivatives showed better antitumor activity than naturally occurring 4-oxa-1-azabicyclo[3.2.0]heptan-7-one derivative G0069A. The trans isomers exhibited superior stability and activity over the cis isomers at the 3- and 5-position. Some of these compounds showed strong cytotoxicity against P388 and KB cells with IC(50) value ranging from 0.004 to 0.6 micro g/ml and they did not show any cross resistance against ADR, 5-FU and VCR resistant cell lines in vitro. Of these, 3-hydroxy methyl, 3-(2-amino-2-carboxy-1-benzyloxy ethyl) and 3-(2-amino-2-carboxy ethyl) 4-oxa-1-azabicyclo[3.2.0] heptan-7-one inhibited 71 84% in vivo tumor growth of colon 26 and S-180 cells subcutaneously implanted into mice at a varying dose between 0.625-15 mg/kg/day depending upon the compounds and the tumor cell lines. PMID- 14529452 TI - The quinolone family: from antibacterial to anticancer agents. AB - The present review focuses on the structural modifications responsible for the transformation of an antibacterial into an anticancer agent. Indeed, a distinctive feature of drugs based on the quinolone structure is their remarkable ability to target different type II topoisomerase enzymes. In particular, some congeners of this drug family display high activity not only against bacterial topoisomerases, but also against eukaryotic topoisomerases and are toxic to cultured mammalian cells and in vivo tumor models. Hence, these cytotoxic quinolones represent an exploitable source of new anticancer agents, which might also help addressing side-toxicity and resistance phenomena. Their ability to bind metal ion co-factors represents an additional means of modulating their pharmacological response(s). Moreover, quinolones link antibacterial and anticancer chemotherapy together and provide an opportunity to clarify drug mechanism across divergent species. PMID- 14529453 TI - Reactions of water-soluble alkylperoxyl radicals and superoxide with DNA, lipoproteins and phospholipid vesicles: the role played by electrostatic forces. AB - The role of electrostatic forces in free radical biology is very important but it is all too often overlooked. The radicals discussed in this review include positively-charged, negatively-charged and neutral water-soluble alkylperoxyls and superoxide. Important scientific insights have been gained by generating these radicals in known quantities by the thermal decomposition of simple, "clean", chemical precursors in the presence of potential bio-targets. For example, the abilities of these radicals to damage double-stranded DNA, a polyanion, are dictated by Coulombic forces with only the positively-charged peroxyls being capable of directly producing single-strand breaks. The Coulombic control of the reactions and reaction rates of water-soluble peroxyl radicals which are so evident with DNA do not manifest themselves with all electrostatically charged bio-targets, e.g., low density lipoprotein (LDL), probably because the charge on the surface of the LDL is not uniformly distributed. PMID- 14529454 TI - Mechanisms of action of DNA intercalating acridine-based drugs: how important are contributions from electron transfer and oxidative stress? AB - Reactive oxygen species (ROS) are produced continuously in living cells as a by product of respiration and other metabolic activity. Some ROS may react with DNA, and in some cases may abstract an electron from the double helix, leading to long range electron transfer (ET) reactions. Thus, the DNA of living cells may be in a continuous state of ET. We consider here whether acridine-based anticancer or antimicrobial drugs, which bind to DNA by intercalation, might either donate electrons to, or accept electrons from, the double helix, thus actively participating in ET reactions. We focus in particular on two acridine-based drugs that have been tested against human cancer in the clinic. Amsacrine is a 9 anilinoacridine derivative that appears to act as an electron donor in ET reactions on DNA, while N-[2-(dimethylamino)ethyl]acridine-4-carboxamide (DACA) may act as an electron acceptor. Such reactions may make important contributions to the antitumor activity of these drugs. PMID- 14529455 TI - Amyloid beta-peptide [1-42]-associated free radical-induced oxidative stress and neurodegeneration in Alzheimer's disease brain: mechanisms and consequences. AB - In addition to synapse loss, neurofibrillary tangles, and neurodegeneration, oxidative stress and amyloid beta-peptide [Abeta] deposition are hallmarks of Alzheimer's disease [AD] brain. Our laboratory coupled these two characteristics of AD into a comprehensive model to account for the synapse loss and neurodegeneration in AD brain. This model combines much of the extant studies on AD and is based on oxidative stress associated with amyloid beta-peptide. This review presents evidence in support of this model and provides insight into the molecular basis of this devastating dementing disorder. PMID- 14529456 TI - Superoxide and post-ischemic liver injury: potential therapeutic target for liver transplantation. AB - Cessation of blood flow to the liver is required during liver transplantation and resectional surgery. A growing body of experimental evidence suggests that restoration of blood flow to the ischemic liver initiates hepatocellular injury which may lead, in some cases, to severe liver injury and graft failure. A large number of studies have implicated reactive oxygen species as potential mediators of post-ischemic tissue injury. Recent developments in genetic engineering as well as chemical modeling, have allowed for the production of novel free radical scavengers including mutated forms of superoxide dismutase (SOD) and low molecular weight SOD mimics with extended circulating half-lives and/or significant membrane permeability's. Application of these newly developed free radical scavengers show promising results in animal models of liver I/R and may become powerful tools in the treatment of post-ischemic liver injury that occurs in liver transplantation. PMID- 14529457 TI - Recent developments on the role of mitochondria in poly(ADP-ribose) polymerase inhibition. AB - Numerous pathophysiological disorders involve some element of oxidative stress and bioenergetic deficit. Poly(ADP-ribose) polymerase-1 (PARP-1) inhibitors have been used recently as a promising new therapeutic strategy aimed at halting the bioenergetic decline associated with oxidative brain insults and other conditions. PARP-1 uses NAD+ as a substrate and is activated during stressful circumstances, mainly in the nucleus. PARP-1 inhibitors are well known for blocking the excessive consumption of NAD+, thereby preserving energy metabolism. But what is the role of mitochondria in this process? Recent investigations have begun to focus on whether mitochondrial function can also be preserved by PARP-1 inhibitors. This review will present some of the latest mechanistic evidence documenting the potential involvement of PARP-1 inhibitors in protecting mitochondrial function and preventing necrosis, apoptosis and mitochondrial calcium cycling. PMID- 14529458 TI - Role of oxidative stress response elements and antioxidants in prostate cancer pathobiology and chemoprevention--a mechanistic approach. AB - Prostate cancer (PC) has become the most frequently diagnosed neoplasm and the second leading cause of cancer-related mortality in men. Its incidence rate has continued to increase rapidly during the past two decades, especially in men over the age of 50 years as they are living longer. The prostate in aging males is highly susceptible to benign and malignant proliferative changes. About two/thirds of all cancers, however, could have been prevented based upon lifestyle choices. The preventative and therapeutic options available to men prone to prostate cancer (both benign and malignant) are limited. How environment, diet and genetics interact to either induce or prevent prostate cancer (PC) is not known. Free radicals, called reactive oxygen species (ROS), play a significant but paradoxical role acting as a "double-edged sword" to regulate cellular processes. Recent in vitro studies using benign prostate hyperplasia (BPH) and PC cell lines grown under various oxidative stress conditions confirm this theory. This manuscript describes key signal transduction mechanisms involved in ROS induced effects on prostate cell growth, cell-cycle checkpoints, apoptosis and transcription factors and the role of potential dietary antioxidants on these mechanisms. It is important to understand underlying signaling mechanisms affected by oxidative stress so as to scientifically prove the efficacy and safety of potential antioxidants in PC prevention. Thus by identifying several potential preventive and therapeutic molecular targets in prostate and by devising better chemo-preventive and chemotherapeutic strategies for controlling PC progression, one can envision significant drop in number of deaths, cut down health care costs and improve the quality of life. PMID- 14529459 TI - Neurotransmission and neurotoxicity by nitric oxide, catecholamines, and glutamate: unifying themes of reactive oxygen species and electron transfer. AB - This review treats the mechanism of nitric oxide, catecholamines, and glutamate as important neurotransmitters and as neurotoxins, based on involvement of reactive oxygen species (ROS) and electron transfer (ET). ROS and ET can serve as a unifying framework for both transmission and toxicity, with ROS concentration being a crucial issue. Cell signaling, electrochemistry, antioxidants, and apoptosis are also discussed. PMID- 14529460 TI - Mechanism of organophosphates (nerve gases and pesticides) and antidotes: electron transfer and oxidative stress. AB - Evidence indicates that nerve gas toxins operate in ways in addition to inhibition of acetylcholine esterase. Alternative bioactivities are discussed with focus on electron transfer. The main class, including pralidoxime (2-PAM), incorporates conjugated iminium and oxime moieties that are electron affinic. Various physiological properties of iminium and oxime species are reviewed. The organophosphates encompass both nerve gases and insecticides, possessing similar properties, but different activities. Toxic manifestations are apparently due, in part, to oxidative stress. Alkylation of DNA takes place which may lead to generation of reactive oxygen species. Structure-activity relationships are examined, including reduction potentials and the captodative effect. PMID- 14529461 TI - Mechanism of drug and toxic actions of gossypol: focus on reactive oxygen species and electron transfer. AB - Gossypol, a constituent of cottonseeds, displays various drug properties, including antifertility and anticancer. Toxicity is shown against the reproductive system, heart, liver, and membranes. The compound exhibits pro- and anti-oxidant behavior. Electron transfer (ET) functionalities, present in gossypol and its metabolites, comprise conjugated dicarbonyl, a quinone derivative, Shiff bases, and metal complexes. The parent possesses a reduction potential favorable for in vivo ET. Considerable evidence points to oxidative stress (OS), formation of reactive oxygen species (ROS), and DNA scission, characteristics of redox cycling by ET in biosystems. Mechanistic aspects are addressed with OS-ROS-ET as the guiding theme, in addition to other modes of action resulting in a multifaceted scenario. PMID- 14529462 TI - Cannabinoids: mechanisms and therapeutic applications in the CNS. AB - Cannabinoids comprise three classes of compounds, the active components of marijuana (Cannabis sativa), as well as endogenous and synthetic derivatives. To date, two distinct cannabinoid receptors (CB1 and CB2) have been discovered, but evidence for further receptor types has been brought forward. The potential use of cannabinoids for medicinal purposes has long been known, but the mechanisms of action of both exogenously applied and endogenous cannabinoids are only partly established. For nervous system disorders, cannabinoids may be useful by modulating neurotransmission and calcium homeostasis as well as by anti inflammatory and anti-oxidant actions. Some cannabinoids can also trigger cell death, which may be of therapeutic benefit in the treatment of malignant tumours. A number of both in vitro and in vivo models have provided promising but diverse evidence for cannabinoid protection in glutamate-mediated excitotoxicity, hypoxia and glucose deprivation, brain trauma, epilepsy and MS. Subsequent to many preclinical investigations, clinical trials are now underway in a variety of the above applications. Overall, the understanding of the therapeutic relevance of cannabinoids will rely on further investigations into the neuroprotective and neurotoxic potency of cannabinoids in animal models and humans, as much as on a further advancement of our general understanding of the endocannabinoid system and the development of specific compounds devoid of unwanted psychoactive side effects. PMID- 14529463 TI - Recent developments in the synthesis and discovery of oligosaccharides and glycoconjugates for the treatment of disease. AB - The development of the first automated oligosaccharide synthesizer, along with new methods for screening carbohydrate ligand arrays is likely to lead to a rapid acceleration in both our ability to synthesize these molecules, and understand the roles of oligosaccharides and glycoconjugates in biology. Consequently we may uncover new avenues for therapeutic intervention more rapidly. These recent developments are very important since our understanding of the role of glycoconjugates in nature has traditionally fallen far behind that of the other biopolymers such as proteins and nucleic acids as the formation of, for example, glycosylated proteins is not template driven. The chemical synthesis of oligosaccharides and glycoconjugates has provided us with new potential cancer vaccines, antibiotics and new biotechnological tools. Glycobiologists have employed many such tools to uncover new signalling roles for oligosaccharides and glycoconjugates. In this review we aim to highlight some emerging methods for glycoconjugate assembly and screening, and discuss innovative approaches to glycoconjugate based drug design and delivery, all of which are, and will continue to be, fruitful avenues for medicinal chemistry research. PMID- 14529464 TI - A multi-functional organelle mitochondrion is involved in cell death, proliferation and disease. AB - The mitochondrion, long considered an organelle specific to energy metabolism, is in fact multi-functional and involved in many diseases. Mitochondrial DNA accumulates somatic mutations during aging, the progression of cancer and diabetes. Most cancer cells contain homoplasmic mutations in the mitochondrial genome. Although little is known about the contributions of mutations to carcinogenesis, some mutations in the nuclear genes encoding mitochondrial proteins have been identified as responsible for certain familial cancers. Mitochondria play an essential role in generating the germ line by releasing mitochondrial ribosomal RNAs, by which the germ line transfers the genetic information necessary for life to the next generation. Collaboration between mitochondria and the cytosol occurs in several metabolic pathways. Many enzymes involved in synthesizing uridine, heme and steroids and in the urea cycle are located inside mitochondria. Notably, a reaction involved in the synthesis of UMP is coupled with the energized state of mitochondria. Thus, the synthesis of DNA and RNA should be indirectly coupled with the energized state of mitochondria. Additionally, storing calcium is an important role of mitochondria. Calcium functions as a second messenger in signal transduction, however, it also activates several proteinases or lipases to induce damage. The mitochondrion plays a significant role in necrosis and is a center for apoptosis, determining its initiation, regulation and execution. Thus, the mitochondrion is widely involved in cell proliferation, cell death and disease. PMID- 14529465 TI - Mitochondrial generation of reactive oxygen species and its role in aerobic life. AB - Mitochondria are the major site for the generation of ATP at the expense of molecular oxygen. Significant fractions (approximately 2%) of oxygen are converted to the superoxide radical and its reactive metabolites (ROS) in and around mitochondria. Although ROS have been known to impair a wide variety of biological molecules including lipids, proteins and DNA, thereby causing various diseases, they also play critical roles in the maintenance of aerobic life. Because mitochondria are the major site of free radical generation, they are highly enriched with antioxidants including GSH and enzymes, such as superoxide dismutase (SOD) and glutathione peroxidase, on both sides of their membranes to minimize oxidative stress in and around this organelle. The present work reviews the sites and mechanism of ROS generation by mitochondria, mitochondrial localization of Mn-SOD and Cu,Zn-SOD which has been postulated for a long time to be a cytosolic enzyme. The present work also describes that a cross-talk of molecular oxygen, nitric oxide (NO) and superoxide radicals regulates the circulation, energy metabolism, apoptosis, and functions as a major defense system against pathogens. Pathophysiological significance of ROS generation by mitochondria in the etiology of aging, cancer and degenerative neuronal diseases is also described. PMID- 14529466 TI - MPP+ analogs acting on mitochondria and inducing neuro-degeneration. AB - This review focuses on the mechanisms of action and the injurious effect of complex I inhibitors, of which 1-methyl-4-phenylpyridinium ion (MPP(+)) is a well studied example. These compounds can be divided into two groups, i.e. competitive inhibitors with respect to ubiquinone, such as piericidine A, and non-competitive inhibitors such as rotenone. Complex I inhibitors such as MPP(+) have been reported to induce anatomical, behavioral, and biochemical changes similar to those seen in Parkinson's disease, which is characterized by nigrostriatal dopaminergic neuro-degeneration. Spectroscopic analyses and structure-activity relationship studies have indicated that the V-shaped structure of the rotenone molecule is critical for binding to the rotenone binding site on complex I. Many isoquinoline derivatives, some of them endogenous, are also complex I inhibitors. Many lines of evidence show that complex I inhibitors elicit neuronal cell death. Recently, it was reported that chronic and systemic exposure to low-dose rotenone reproduces the features of Parkinson's disease. This work further focused attention on compounds acting on mitochondria, such as MPP(+). In Guadeloupe, the French West Indies, patients with atypical parkinsonism or progressive supranuclear palsy are frequently encountered. These diseases seem to be associated with ingestion of tropical herbal teas or tropical fruits of the Annonaceae family, which contain complex I inhibitors such as benzylisoquinoline derivatives and acetogenins. Complex I inhibitors may not simply result in reactive oxygen species generation or ATP exhaustion, but may influence complex downstream signal transduction processes. An understanding of these changes would throw light on the ways in which complex I inhibitors induce a wide range of abnormalities. PMID- 14529467 TI - Mitochondria-targeting therapeutic strategies for overcoming chemoresistance and progression of cancer. AB - The cytotoxic effects of many anticancer drugs are mediated via the apoptotic pathways. Chemoresistant tumor cells have acquired the ability to evade the action of multiple classes of anti-cancer drugs. One mechanism by which tumor cells survive in the presence of chemotherapy is by increasing anti-apoptotic activities. Since mitochondria are critical 'gatekeepers' to the apoptosis process, development of cytotoxic drugs that target mitochondria may provide a new strategy to induce apoptosis in tumor cells. Mitochondrial permeability transition pore complex (PTPC) controls mitochondrial membrane permeabilization, which is a critical event in the process leading to chemotherapy-induced apoptosis. Therefore, targeting of PTPC components may overcome chemoresistance in tumor cells. Moreover, alterations in mitochondrial DNA such as mutation and the subsequent dysfunction of mitochondrial respiratory enzyme have been reported in various types of cancer, and their functional consequences are associated cancer development, chemoresistance, and therapeutic implications. In this mini review, we aim to provide a brief review on several mitochondria-targeting strategies to overcome chemoresistance in cancer. PMID- 14529468 TI - Medicinal and genetic approaches to the treatment of mitochondrial disease. AB - Although great progress has been made in our understanding of the molecular bases of mitochondrial disorders due to defects in the respiratory chain, little exists in the way of rational therapy. Possible therapeutic approaches include: palliative therapy; removal of noxious metabolites; administration of artificial electron acceptors, metabolites, and free radical scavengers; genetic counseling; and gene therapy. There has been progress with each of these approaches, although much work remains to be done. Finally, a novel approach to treating a specific mitochondrial disorder, MELAS, is presented. PMID- 14529469 TI - Parasite mitochondria as drug target: diversity and dynamic changes during the life cycle. AB - Parasites have developed a wide variety of physiological functions to survive within the specialized environments of the host. Regarding energy metabolism, which represents an essential factor for survival, parasites adapt low oxygen tension in host mammals using metabolic systems that differ substantially from those of the host. Most parasites do not use free oxygen available within the host, but employ systems other than oxidative phosphorylation for ATP synthesis. Furthermore, parasites display marked changes in mitochondrial morphology and components during the life cycle, and these represent very interesting elements of biological processes such as developmental control and environmental adaptation. The enzymes in parasite-specific pathways offer potential targets for chemotherapy. Cyanide-insensitive trypanosome alternative oxidase (TAO) is the terminal oxidase of the respiratory chain of long slender bloodstream forms of the African trypanosome, which causes sleeping sickness. Recently, the most potent inhibitor of TAO to date, ascofuranone, was isolated from the phytopathogenic fungus, Ascochyta visiae. The inhibitory mechanisms of ascofuranone have been revealed using recombinant enzyme. Parasite-specific respiratory systems are also found in helminths. The NADH-fumarate reductase system in mitochondria form a final step in the phosphoenolpyruvate carboxykinase (PEPCK)-succinate pathway, which plays an important role in anaerobic energy metabolism for the Ascaris suum adult. Enzymes in this system, such as NADH rhodoquinone reductase (complex I) and rhodoquinol-fumarate reductase (complex II), form promising targets for chemotherapy. In fact, a specific inhibitor of nematode complex I, nafuredin, has been found in mass-screening using parasite mitochondria. PMID- 14529470 TI - Perspectives on cancer therapy: cell cycle blockers and perturbators. AB - Over the past few decades, remarkable advances have been achieved in cancer therapy, including chemotherapeutic agents, their mode of application and more broader therapeutic strategies. Promising new therapeutic targets have emerged in the past ten years as a result of recent advances in our understanding of the pathobiology of malignant cells, in particular, regarding functions of suppressor oncogene products. Among them, the agents that alter the cell cycle have recently been of particular interest, since cell cycle regulation is basic mechanism underlying cell fate, i.e., proliferation, differentiation or death. Furthermore, the human genome project has made possible the future development of so-called "tailor-made medicine", i.e. the design of appropriate drugs for specific genetic profiles and application of drugs that are tailored to each tumor and patient. In this article, we will introduce and discuss recent progress in the development of agents that influence the cell cycle and their future potential in cancer therapy from three standpoints with our own experimental works; i) the inhibition of cell proliferation and/or induction of differentiation by cyclin-dependent kinase (cdk)-inhibitor, e.g. olomoucin, butyrolactone-I, ii) induction of apoptosis by directing "abortive cell cycle", or the transient upregulation of cdk activity, e.g. flavopiridol, and iii) countering the development of drug resistance by adjunctive administration of cdk-inhibitors with conventional anti-cancer drug, e.g., p21-gene transfer with cisplatin. Conclusively none of these three approaches by itself is satisfactory, and that the effective cancer therapies will require the administration of several agents and/or methods under the design of their synergistic effects. PMID- 14529471 TI - Natriuretic peptides in alcohol withdrawal: central and peripheral mechanisms. AB - Abrupt cessation of long-term alcohol consumption produces well-defined symptoms called alcohol withdrawal (AW). The exact pathophysiological mechanisms involved in the appearance of AW symptoms and particularly those related to the precipitation of delirium tremens (DT), still await clarification in spite of the fact that the prediction of complicated AW is essential to guarantee that appropriate therapies may be planned in advance. Changes in central nervous system (CNS) glutamate- and GABA-transmission and a role of voltage-operated calcium channels are equally important elements of neuroadaptation to the chronic presence of alcohol. In addition to the CNS regulation, however, changes in peripheral fluid and electrolyte homeostasis may accompany, and are expected to modify the clinical symptoms of AW. In an early phase of acute withdrawal, plasma levels of atrial natriuretic peptide (ANP), plasma renin activity and aldosterone are high. In patients with DT, elevated levels of ANP were observed days before the actual onset of DT. It is concluded that the altered plasma ANP secretion might be associated with, and therefore used as an indicator of the onset of DT. However, ANP is present in and produced by the brain and thus it can be regarded as a neuropeptide. The role of CNS ANP was studied in mice, rendered tolerant to and physically dependent on alcohol. Intracerebroventricular injections of ANP attenuated, whereas those of an antiserum against ANP intensified hyperexcitability during AW. ANP in the brain - the content of which undergoes sensitive changes in the hippocampus during AW appears to interact primarily with glutamate transmission through the NMDA-receptors. This brain structure is of utmost importance for the generation of withdrawal-related hyperexcitability. It is concluded that peripheral secretion of ANP might be a diagnostics indicator, whereas ANP in the CNS might be a modulator of AW. PMID- 14529472 TI - Altered protein acetylation in polyglutamine diseases. AB - Polyglutamine diseases are hereditary neurodegenerative disorders caused by the expansion of a CAG repeat in the disease gene. A dominant gain of function is associated with these expanded alleles. The resulting elongated polyglutamine repeats are thought to cause structural changes in the affected proteins, leading to aberrant interactions such as those that allow formation of extra- and intranuclear aggregates. However, self-association is not the only interaction the polyglutamine domain is capable of mediating. Many cellular proteins can be sequestered into inclusions or bound by more soluble forms of the mutant proteins. One group of proteins that binds to and whose activity may be altered by polyglutamines is Histone Acetyltransferases (HATs). HATs are responsible for the acetylation of histones and several other important proteins and this modification results in altered function of the target protein. HATs regulate cellular processes at levels as different as modifying transcriptional competence of chromosomes, temporal regulation of promoter activity and protein activation / inactivation. Recent studies show that the altered balance between protein acetylation and deacetylation may be a key process contributing to expanded polyglutamine-induced pathogenesis. The restoration of this balance is possible by the genetic or pharmacological reduction of the opposing enzyme group, i.e. the Histone Deacetylases (HDACs). Recent progress in HDAC research has made the development of inhibitors of specific HDAC family proteins possible and these compounds could be effective candidates for treatment of these devastating diseases. PMID- 14529474 TI - Intracellular thiol concentration modulating inflammatory response: influence on the regulation of cell functions through cysteine prodrug approach. AB - Oxidative stress is defined as the consequence of overpowering of the immune system's reaction, which causes increased production of the reactive oxidative species (ROS) greater than the antioxidant protection. Tissue injury and oxidation of the circulating molecules may be the consequences. Moreover, the sulphur-containing amino acids (SAA) fate is perturbed during stress. The altered biochemical rules during inflammation weaken the anti-oxidant mechanism, and the extra-supply of SAA under inflammatory conditions can help to restore homeostasis. In brief, the main biochemical steps during inflammation are: The production of Cytokines, Acute Phase Protein, and Glutathione (GSH) pool are strongly modified during inflammation. The GSH participates in many important physiological processes controlling the homeostasis of the cells. A higher demand of Cysteine (Cys) supply causes difficulties in maintaining a constant GSH level. The role of GSH as a key regulator of thiol redox intracellular balance is established. This reveals that GSH is essential in regulating the cell's life cycle and that the reduction of intracellular GSH contributes to chronic inflammation. The fact that Cys availability is generally a limiting factor for the GSH synthesis stimulated the development of a pharmacologically useful Cys pro-drug. The simplest derivative is N-acetylcysteine (NAC), which appears to be the prototype of all Cys suppliers. Different approaches are presented here. PMID- 14529475 TI - Muscarinic and nicotinic cholinergic agonists: structural analogies and discrepancies. AB - Acetylcholine, the first identified neurotransmitter acts on both types of cholinergic receptors. Both rigid and flexible derivatives of acetylcholine could either be selective muscarinic or selective nicotinic agonists while some compounds show activity at both receptor subclasses. Earlier structure-activity considerations are revisited. Ligand and receptor based calculations have been applied in the hope to identify characteristic geometrical and steric requirements for the activity on the receptor subtypes. Results are treated critically and applied cautiously for predicting selective structural requirements by the cholinergic receptor subclasses. PMID- 14529473 TI - Ribozyme-based gene-inactivation systems require a fine comprehension of their substrate specificities; the case of delta ribozyme. AB - The ability of ribozymes (i.e. RNA enzymes) to specifically recognize and subsequently catalyze the cleavage of an RNA substrate makes them attractive for the development of therapeutic tools for the inactivation of both viral RNAs and mRNAs associated with various diseases. Several applicable ribozyme models have been tested both in vitro and in a cellular environment, and have shown significant promise. However, several hurdles remain to be surpassed before we generate a useful gene-inactivation system based on a ribozyme. Among the most important requirements for further progress are a better understanding of the features that contribute to defining the substrate specificity for cleavage by a ribozyme, and the identification of the potential cleavage sites in a given target RNA. The goal of this review is to illustrate the importance of both of these factors at the RNA level in the development of any type of ribozyme based gene-therapy. This is achieved by reviewing the recent progress in both the structure-function relationships and the development of a gene-inactivation system of a model ribozyme, specifically delta ribozyme. PMID- 14529476 TI - Aurones: a subclass of flavones with promising biological potential. AB - Aurones [2-benzylidenebenzofuran-3(2H)-ones] are the secondary metabolites natural compounds belong to the flavonoids family, and structurally are the isomers of flavones, widely present in fruits and flowers where they play significant role in the pigmentation of the part of plant in which they occur. Literature survey clearly indicates that flavones, chalcones, flavonols and isoflavones have been studied largely for their therapeutical potential. Somehow, aurones still are less studied and it is only recently that these compounds have begun to be investigated. In this review, we report the recent advances made on the therapeutical potential of aurones in different biological areas. Their synthesis, structure-activity relationships, the importance of the substitution pattern will also be discussed. Finally, some aspects regarding the possible development of aurones will be highlighted briefly. PMID- 14529477 TI - Histone deacetylase inhibitors: from chromatin remodeling to experimental cancer therapeutics. AB - Histone deacetylases (HDACs) are key enzymes in the regulation of gene expression. By maintaining the dynamic equilibrium of the acetylation status of highly conserved lysine residues on histones, they regulate chromatin remodeling and gene expression. A link between aberrant HDAC activity and cancer has been widely reported and HDAC inhibitors have been shown to inhibit the proliferation of human tumor cell lines in vitro. Furthermore, several HDAC inhibitors have exhibited potent anti-tumor activity in human xenograft models, suggesting this class of compounds to be promising novel cancer therapeutic agents. This review provides an update on the current knowledge of HDAC inhibition with a focus on the most recent progress of HDAC inhibitors in clinical development. PMID- 14529478 TI - From discovery to the coming generation of histone deacetylase inhibitors. AB - Trichostatin A (TSA) is a Streptomyces metabolite that causes differentiation of murine erythroleukemia cells as well as specific inhibition of the cell cycle of some lower eukaryotes and mammalian cells. The targeted molecule of TSA has been shown by genetic and biochemical analyses to be histone deacetylases (HDACs). Histone acetylation is a key modification to control transcription, and HDACs are profoundly involved in pathogenesis of cancer through removing acetyl groups from histones and other transcriptional regulators. Trapoxin (TPX) and FK228 (also known as FR901228 and depsipeptide because FK228 = FR901228 = depsipeptide), structurally unrelated microbial metabolites, were also shown to inhibit HDACs. These HDAC inhibitors cause cell cycle arrest, differentiation and/or apoptosis of many tumors, suggesting their usefulness for chemotherapy and differentiation therapy. In addition, HDAC inhibitors play important roles in identifying the specific function of the enzymes. Indeed, we identified tubulin as one of the substrates of HDAC6 by means of differential sensitivity to HDAC inhibitors. Since recent studies have revealed that HDACs are structurally and functionally diverse, it should be important to develop inhibitors specific to individual enzymes as more promising agents for cancer therapy. We have synthesized novel TSA/TPX hybrids, which will serve as a basis for developing enzyme-specific HDAC inhibitors. PMID- 14529479 TI - Novel hydroxamate and anilide derivatives as potent histone deacetylase inhibitors: synthesis and antiproliferative evaluation. AB - There is a currently growing interest in the development of histone deacetylase inhibitors (HDACs) as anticancer agents. Histone deacetylases are critically important in the functional regulation of gene transcription as well as in chromatin structure remodeling. A number of small molecule inhibitors of HDAC, such as the naturally occurring trichostatin A (TSA), as well as synthetic compounds, such as suberoylanilide hydroxamic acid (SAHA), scriptaid, oxamflatin or MS-275, have been reported to induce differentiation of several cancer cell lines and suppress cell proliferation. This article will review the recent progress being made in our laboratories in the development of two new families of potent HDAC inhibitors: sulfonamide hydroxamic acids and anilides, as well as TSA like straight chain derivatives. Some of these compounds inhibit partially purified recombinant human HDAC enzymes with IC(50)'s in the micromolar to low nanomolar range and can induce hyperacetylation of histones in human cancer cells. These compounds significantly inhibit proliferation, induce expression of p21(WAF1/Cip1), and cause cell cycle arrest in various human cancer cells. The lead candidates were screened in a panel of human tumor and normal cell lines. The inhibition of HDAC activity represents a novel approach for intervening in cell cycle regulation and may be used in future cancer therapies. The structure activity relationships, the antiproliferative activity and the in vivo efficacy are discussed. PMID- 14529481 TI - The discovery of NVP-LAQ824: from concept to clinic. AB - The natural products trapoxin B and trichostatin A, as well as the novel marine natural product psammaplin A (PSMA) were found in a cell-based screen for compounds that induced the expression of the cyclin dependent kinase inhibitor p21(waf1). The mechanism of p21(waf1) induction for these compounds was via histone deacetylase (HDAC) inhibition. Of these compounds, PSMA was of interest because of its novel structure, but the physiological stability of it, and its analogs was poor. Thus, a directed medicinal chemistry effort was undertaken to prepare analogs of the simple HDAC inhibitor dimethylaminobenzamidylcaprylic hydroxamate (DBCH), which led to chromenone amide 9. This compound was efficacious in the HCT116 colon xenograft assay, but was difficult to formulate in pharmaceutically acceptable vehicles. In parallel with these efforts, a screen of the Novartis compound archive for novel HDAC inhibitors uncovered the cinnamyl hydroxamic acid NVP-LAK974. This compound had good enzyme and cellular potency, but poor efficacy in vivo. A systematic structural exploration of cinnamyl hydroxamates based on NVP-LAK974 was undertaken with the goal of finding a novel, well-tolerated and efficacious HDAC inhibitor. Several derivatives were found to be efficacious in the xenograft assay. Of those compounds, NVP-LAQ824 distinguished itself due to its tolerability, efficacy and potency. Based, in part, on these properties, NVP-LAQ824 is currently undergoing human clinical trials as a novel anti-cancer agent. PMID- 14529480 TI - Histone deacetylase inhibitors: the Abbott experience. AB - Histone deacetylase inhibitors have generated significant interest as anti-cancer agents due to their ability to cause growth arrest, terminal differentiation and/ or apoptosis in carcinoma cells. Abbott entered this area after the serendipitous discovery of the biaryl hydroxamate A-161906 in a TGF beta mimetic screen and the subsequent identification of this compound as an inhibitor of selected HDACs. The complex biology of these enzymes became evident when cloning and expression of the HDACs demonstrated that they were present as multiprotein and, in some cases, multi-HDAC containing complexes in their active forms. This discovery suggested that any selectivity determinations would have to be considered in the context of these multi-protein/HDAC complexes. However, siRNA gene knockdown studies did demonstrate that reduction of the Class I HDACs resulted in a phenotype similar to that observed with small molecule HDAC inhibitors. Evaluation of the Abbott small molecule HDAC inhibitors utilized a Class I HDAC (HDAC 1/2) preparation and antiproliferation assays using HT1080 fibrosarcoma and MDA435 breast carcinoma cells. Characterization of several series of hydroxamic acids indicated that while many of these analogs possessed potent enzymatic and cellular activity, in general these compounds had unacceptable pharmacokinetic profiles and marginal antitumor effects. Replacement of the potentially labile hydroxamic acid moiety with a trifluoromethyl ketone or a ketooxazole gave measurable HDAC potency but only modest cellular and in vivo activity. However, hydroxamate replacement with an alpha-ketoamide moiety provided potent HDAC inhibitors (IC(50) values as low as 3 nM) with excellent cellular activity (IC50 values < 0.2 microM) and measurable anti-tumor activity in a flank tumor growth model. PMID- 14529482 TI - Microwave-assisted polymer-supported combinatorial synthesis. AB - Lead identification and optimization is always a challenge to the medicinal chemists in drug discovery. Numbers of simple to complex and smaller to bigger organic compounds are prepared to meet the screening purpose of biological targets. Conventional solution phase synthetic methodologies are lacking the speed to run along with the need of medicinally interesting compounds due to their long reaction time, tedious work-up and purification problems. Alternatively opted polymer-supported synthesis of combinatorial libraries has been emerged as a promising tool in generating large numbers of structurally diverse molecules in a manner rapid and parallel. Microwave-assisted solid/liquid phase combinatorial synthetic techniques have been proved efficient in reducing the reaction time from days & hours to minutes & seconds and more promisingly to produce improved yields with high purities. This review briefs about the theory behind microwave chemical technology and glimpses of recent advancements in its application on polymer supported combinatorial synthesis. PMID- 14529483 TI - Bilayer-forming synthetic lipids: drugs or carriers? AB - Since their introduction as bilayer-forming synthetic compounds in the eighties, dioctadecyldimethylammonium (DODA) and dihexadecylphosphate (DHP) salts have found many uses in strategic, applied areas. In particular, DODA chloride or bromide vesicles interacted with negatively charged prokaryotic or eukaryotic cells, yielding adsorption isotherms of high affinity for the cell surface, causing cell adhesion and flocculation, changing the cell surface charge from negative to positive, and causing loss of cell viability over DODA concentration ranges that depended on the cell type being tested. This work reviews data on DODA effects on cell viability (bacteria, fungus and cultured mammalian cells) to propose DODA salts as effective anti-microbial agents that exhibit differential cytotoxicity in vitro and, therefore, deserve to be investigated as potential drugs. The full utility of these inexpensive synthetic bilayers and bilayer fragments able to act as drugs themselves and, simultaneously, as drug, gene or vaccine carriers remains hitherto unexplored. PMID- 14529484 TI - Approaches to improve cellular retention of radiohalogen labels delivered by internalising tumour-targeting proteins and peptides. AB - Specific targeting of radionuclides is a promising approach to improve diagnosis and treatment of tumors. Targeting vectors may be monoclonal antibodies directed toward tumour-specific antigens or regulatory peptides binding to receptors overexpressed on or by malignant cells. Depending on the aim of the procedure and the biokinetics of the targeting vectors, radionuclides with different nuclear properties (decay scheme, half-life, etc.) must be applied. Halogen radioisotopes are attractive since they exhibit a variety of nuclear properties suitable for various applications. At the same time, their chemistry shows great similarities, which enables the use of similar labelling procedures for different nuclides. A problem in using radiohalogens for labelling of tumour-targeting proteins and peptides is that the commonly used radiohalogenation methods provide labels, which, after internalisation and lysosomal digestion, rapidly "leak" from malignant cells as radiohalogenated degradation products. The main reason for such leakage is free diffusion of the radiometabolites through lysosomal and cellular membranes. This review describes current approaches in molecular design to improve cellular retention of radiohalogen labels. These approaches include the use of prosthetic groups for the attachment of radiohalogens to targeting vectors of bulky hydrophilic non-charged molecules, molecules positively charged at lysosomal pH and negatively charged molecules. The emphasis in this paper is on labelling chemistry and the results of the biological testing of labelled compounds. PMID- 14529485 TI - The von Hippel-Lindau gene and protein in tumorigenesis and angiogenesis: a potential target for therapeutic designs. AB - The von Hippel-Lindau (VHL) protein is able to suppress tumor growth and to down regulate many angiogenic factors, and is ubiquitously detected in adult and fetal tissues. This makes VHL an excellent target for therapeutic intervention. Observation of VHL alterations in sporadic tumors has been increasing as a result of examination of abnormalities other than intragenic mutations. These abnormalities include loss of chromosome 3p25, changes in the promoter, down regulation of transcript, and changes in protein level. This article also presents the finding of differential expression of two common VHL proteins among rat tissues, suggesting tissue- and development-dependent functional difference between these two isoforms. Molecular pathways linking VHL to angiogenesis have been extensively characterized and mechanisms have been proposed to explain how altered VHL leads to tumorigenesis. VHL functions in the presence of oxygen and/or oxygen species. Two strategies are proposed here for anti-tumor and anti angiogenic treatments of VHL-deficient tumors and those without detectable VHL intragenic mutations. One is to restore wild-type VHL function in VHL-deficient tumors and the other is to enhance wild-type VHL expression and activity in tumors under hypoxic conditions. PMID- 14529487 TI - Obtaining site-specific calcium-binding affinities of calmodulin. AB - Calmodulin (CaM) is an EF-hand Ca(II)-binding protein involved in the regulation of many important biological processes. To date, there is a wealth of information available concerning studies to obtain site-specific calcium binding affinities of CaM, and further to estimate the cooperativity of calcium binding using mutational studies, peptide models, and proteolytic fragmentation. In this paper, we will discuss the energetics of calcium binding and the strong relationship between calcium binding cooperativity and conformational change. We then explain the difficulty of studying key determinants of calcium binding affinity of CaM due to the large change of calcium binding affinity upon mutation. Subsequently, we will introduce "grafting" as a novel approach to obtain the site-specific metal binding properties of calmodulin. PMID- 14529488 TI - An antiparallel beta-sheet and a beta-turn characterize the structure of antiviral HIV-1 peptide T140, as revealed by 2D NMR and MD Simulations. AB - The polyphemusins present in the hemocytes of the horsechoe crab and their structurally modified analogs have been shown to exhibit activity against HIV-1. Among the many variants, T22 ([Tyr(5,12), Lys(7)]-polyphemusin II), and its shorter and more potent analog, T140 [Arg(1)-Arg-2-Nal-Cys-Tyr(5)-Arg-Lys-D-Lys Pro-Tyr(10)-Arg-Cit-Cys-Arg(14)] (Polyphemusin II-derived peptide), affect the HIV-cell fusion process and inhibit the T-cell line-tropic (T-tropic) HIV-1 infection. Conformational studies of polyphemusin II derived peptide have been carried out by (1)H and (13)C 2D-NMR and MD simulations in water and HFA (40%). The NMR parameters of chemical shift, temperature coefficients of the NH chemical shifts, (3)JNHalpha coupling constants and the pattern of nOe's were used to deduce the structural characteristics. Solution structures were generated using dihedral and distance restraints by MD simulations. The structures are characterized by a dominant family possessing an anti-parallel beta-pleated sheet that is constrained by the disulphide bridge between Cys4 and Cys13. The two strands of the beta-sheet are joined by a Type II' beta-turn spanning the residues Lys(7)-D-Lys(8)-Pro(9)-Tyr(10). This conformation is present in both water and HFA. The only difference in the two structures is that the beta-strands are more cohesive in HFA being firmly held by H-bonds. The solution structures generated from MD simulations were refined by MARDIGRAS to R-factors of 0.44 and 0.57 in water and HFA respectively. The conformation deduced for T140 is very similar to that reported for T22 and is thought to be associated with their anti HIV activity. PMID- 14529489 TI - Mutation of the hydrophobic residue on helix alpha5 of the Bacillus thuringiensis Cry4B affects structural stability. AB - Cry4B toxin is a mosquito-larvicidal protein from the Bacillus thuringiensis subsp. israelensis. We have investigated the role of two conserved hydrophobic residues of Cry4B in structural stabilization. Substitutions of the leucine-175 and isoleucine-189 on helix alpha5 with valine and leucine did not affect the expression level, solubility and proteolytic processing. Steady state analysis of an unfolding experiment as monitored by circular dichroism and fluorescence spectroscopy demonstrated a typical two-state transition. The determined unfolding free energy for the L175V mutant revealed a structural destabilization of 10.49 kcal/mol relative to the wild type. However unfolding kinetic analysis gave identical activation energy for wild type and both mutants. Our findings suggested that a perturbation on the close packing of the hydrophobic side chains in protein interior could lead to a significant destabilization of the native conformation. PMID- 14529490 TI - Protein production by stationary phase induction (SPI). AB - An alternative method for expressing the recombinant proteins in Escherichia coli is proposed. Unlike the ordinary induction protocol where the cells in the early- to mid-log phase are induced for the protein production, this alternative protocol utilizes the cells in the stationary phase. By using a glutathione S transferase fusion protein as an example, the protocol proposed in this report yielded a higher amount of the desired protein than that from the ordinary protocol. This protocol also suppressed the proteolytic cleavage of the desired protein in the Escherichia coli cytoplasm, thus it should be particularly useful to produce proteins that undergo unwanted cleavages. PMID- 14529492 TI - Proliferative activity of neokyotorphin-related hemoglobin fragments in cell cultures. AB - alpha-Hemoglobin fragments alpha-(133-141), alpha-(134-141), alpha-(135-141), alpha-(137-141), alpha-(134-140), alpha-(133-138), alpha-(134-140) and alpha-(137 138) stimulate L929 tumor cell proliferation, alpha-(134-141) being the most active. alpha-(134-141) stimulates proliferation of M3 melanoma cells, murine embryonic fibroblasts, primary cultures of red bone marrow and spleen cells. In L929 cells the effect of alpha-(134-141) is cell density independent; in M3 cells alpha-(137-141) and alpha-(134-141) are most active at density 10,000 cells/well (96 well plate) independently on FBS content. PMID- 14529491 TI - A non-active site residue, cysteine 69, of glutathione S-transferase ADGSTD3-3 has a role in stability and catalytic function. AB - The Cys69 residue of an Anopheles dirus glutathione S-transferase isoform (adGSTD3-3), was characterized to elucidate its contribution in both catalysis and structural support. Nine mutants were generated at this position by replacing the residue with polar, non-polar and charged residues. The polar residues changed the Vm of the enzymes. With non-polar residues, the enzymes were unable to fold and were expressed in the insoluble inclusion form. With charged residues, the soluble enzyme yields were only 3% of the wild type protein. Molecular dynamics simulation also was performed to understand the changes in the enzyme structure. These findings are additional evidence of the importance of structural residues that affect the enzymatic properties such as Vm, Km and enzyme specificity. PMID- 14529486 TI - Glucagon-like peptide-1 synthetic analogs: new therapeutic agents for use in the treatment of diabetes mellitus. AB - Glucagon-like peptide-1-(7-36)-amide (GLP-1) is a potent blood glucose-lowering hormone now under investigation for use as a therapeutic agent in the treatment of type 2 (adult onset) diabetes mellitus. GLP-1 binds with high affinity to G protein-coupled receptors (GPCRs) located on pancreatic beta-cells, and it exerts insulinotropic actions that include the stimulation of insulin gene transcription, insulin biosynthesis, and insulin secretion. The beneficial therapeutic action of GLP-1 also includes its ability to act as a growth factor, stimulating formation of new pancreatic islets (neogenesis) while slowing beta cell death (apoptosis). GLP-1 belongs to a large family of structurally-related hormones and neuropeptides that include glucagon, secretin, GIP, PACAP, and VIP. Biosynthesis of GLP-1 occurs in the enteroendocrine L-cells of the distal intestine, and the release of GLP-1 into the systemic circulation accompanies ingestion of a meal. Although GLP-1 is inactivated rapidly by dipeptidyl peptidase IV (DDP-IV), synthetic analogs of GLP-1 exist, and efforts have been directed at engineering these peptides so that they are resistant to enzymatic hydrolysis. Additional modifications of GLP-1 incorporate fatty acylation and drug affinity complex (DAC) technology to improve serum albumin binding, thereby slowing renal clearance of the peptides. NN2211, LY315902, LY307161, and CJC-1131 are GLP-1 synthetic analogs that reproduce many of the biological actions of GLP 1, but with a prolonged duration of action. AC2993 (Exendin-4) is a naturally occurring peptide isolated from the lizard Heloderma, and it acts as a high affinity agonist at the GLP-1 receptor. This review summarizes structural features and signal transduction properties of GLP-1 and its cognate beta-cell GPCR. The usefulness of synthetic GLP-1 analogs as blood glucose-lowering agents is discussed, and the applicability of GLP-1 as a therapeutic agent for treatment of type 2 diabetes is highlighted. PMID- 14529493 TI - Inhibitory and promotive effects of polyamines on transglutaminase-induced protein polymerization. AB - Transglutaminase (TGase) has been reported to be involved in the regulation of cell growth. We examined the effects of polyamines on TGase activity. The polymerization of casein was inhibited by putrescine (PUT) and spermidine (SPD). On the other hand, polymerization of N,N-dimethylcasein was increased by spermine (SPM) and SPD. These results suggested polyamines played two distinct roles as inhibitor and promoter for TGase-catalyzed protein polymerization. PMID- 14529494 TI - Production and purification of recombinant human oxytocin overexpressed as a hybrid protein in Escherichia coli. AB - The plasmid DNA pERilox4 containing the gene of the recombinant protein, which included the leader sequence and the oxytocinoyl lysine tetramer, was constructed. The high level of gene expression in E. coli was achieved. The method for purification of the recombinant protein and its isolation in the soluble form was developed. The conditions for digestion of the hybrid protein by trypsin and carboxypeptidase B were matched. The effective method for transformation of oxytocinic acid to oxytocin was worked out. The scheme suggested allowed obtaining oxytocin in high yield. PMID- 14529495 TI - Crystallization of the terminal oxygenase component of biphenyl dioxygenase derived from Rhodococcus sp. strain RHA1. AB - The terminal oxygenase component of the biphenyl dioxygenase (BphA1A2 complex) was over-expressed with a novel over expression system in recombinant Rhodococcus strain and purified. The purified enzyme has been crystallized by the hanging drop vapor diffusion method and subjected to X-ray diffraction analysis. The crystals belong to the tetragonal system in the space group P4(1)2(1)2 or P4(3)2(1)2 and diffract to better than 2.2A resolution. PMID- 14529496 TI - Crystallization and X-ray analysis of NH3-dependent NAD+ synthetase from Helicobacter pylori. AB - The ubiquitous NAD(+) synthetase catalyzes the key step in the biosynthesis of nicotinamide adenine dinucleotide. NH3-dependent NAD(+) synthetase from Helicobacter pylori was purified to homogeneity and crystallized using PEG 1500 as a precipitant. The crystal diffracted up to a resolution of 2.3+ and was found to belong to space group C2 with unit cell dimensions of a = 93.8, b = 48.3, c = 64.2 A and alpha = gamma = 90, beta = 110.0 degrees. PMID- 14529497 TI - Crystallization and preliminary X-ray diffraction study of hemoglobin D from the Aldabra giant tortoise, Geochelone gigantea. AB - Hemoglobin D (Hb D) from the Aldabra giant tortoise, Geochelone gigantea, was crystallized by the hanging drop vapor diffusion technique with a precipitant solution containing 10% polyethylene glycol 3350 and 50 mM HEPES-Na, pH 7.5. The Hb D crystals of G. gigantea, which diffract to at least a 2.0 A resolution, belong to the monoclinic space group C2 with unit cell dimensions of a = 112.1 A, b = 62.4 A, c = 54.0 A, and beta = 110.3 degrees. One alphabeta dimer molecule of Hb D existed in an asymmetric unit, with a calculated value of Vm of 2.77 A(3)Da( 1). PMID- 14529498 TI - Physicochemical effects in the representation of molecular structures for drug designing. AB - After the identification of a biological target, drug design is to analyze the relationships between the structure of potential ligands and their biological activity. A hierarchy of structure representation is presented here considering either the constitution of a molecule, its 3D structure, or the molecular surface. At each level, a variety of physicochemical effects can be accounted for. Furthermore, the special requirements of learning algorithm, such as neural networks, are taken into consideration. Application to problems from combinatorial chemistry, lead identification, high-throughput screening, and prediction of ADME-Tox properties are given. PMID- 14529499 TI - Fragmental methods in the analysis of biological activities of diverse compound sets. AB - The current mini-review explains how fragmental methods (FMs) can be used in the analysis and prediction of physicochemical properties and biological activities. The considered properties include log P, solubility, pK(a), intestinal permeability, P-gp substrate specificity and toxicity. The focus will be a description of a "mechanistic" approach, which implies a gradual reduction of alternative explanations for any property or activity. This means a flexible construction of fragmental parameters using large amounts of experimental data. Since biological activities involve multiple (unknown) target macromolecules with multiple binding modes, a stepwise classification (C-SAR) analysis is most useful. It involves the following procedures: (i). construction of physicochemical profiles using parameters that can be reliably predicted, (ii). identification of reactive functional groups and the largest active skeletons, (iii). generalization of these groups and skeletons in terms of "site-specific physicochemical profiling". This entails a dynamic construction of 2D pharmacophores that can be converted into 3D models. PMID- 14529500 TI - The WWW as a tool to obtain molecular parameters. AB - This article analyses molecular property calculation resources available on the Internet. The first section summarizes the on-line database resources that could be useful to search molecular and biological properties of chemicals, and indicates some principal databases with physicochemical, thermochemical, toxicity, cancer and HIV data. The second section overviews popular standalone programs for calculation of molecular descriptors. Some of these programs can be downloaded for free and used as standalone applications for calculation of molecular descriptors. The third section describes on-line tools for the prediction of molecular properties, activities and calculation of molecular descriptors. Analysis of emerging tools that can be useful to developing new on line servers for the prediction of molecular parameters and properties is also given. PMID- 14529501 TI - A comparison of calculated and experimental parameters as sources of structural information: the case of lipophilicity-related descriptors. AB - This review is organized in three parts: firstly there is a general overview of recent developments in lipophilicity written to induce medicinal chemists to question what they want to obtain from this kind of study; secondly, the state-of the-art of experimental and computational determination of log P is briefly reviewed; finally, some applications are discussed to illustrate how much information can be extracted from lipophilicity, and to highlight the difficulty of obtaining a reliable, general method to work with. PMID- 14529502 TI - Applied introduction to multivariate methods used in drug discovery. AB - The number of articles concerning optimization and applications of multivariate techniques in drug discovery testifies the growing importance attributed to these methods. This mini review focuses on some of the basic and most employed multivariate techniques in drug discovery research. Examples from the literature were selected to illustrate a number of potential applications. PMID- 14529503 TI - The use of molecular descriptors in the design of gadolinium (III) chelates as MRI contrast agents. AB - Nuclear Magnetic Resonance Imaging (MRI) is a very useful tool in modern medical diagnostics, especially when gadolinium(III)-based contrast agents are administered to the patient with the aim of increasing the image contrast between normal and diseased tissues. The main purpose of this review is to show that a new generation of these contrast agents could be developed by making greater use of soft modelling techniques such as QSAR/QSPR after a suitable description of their molecular structure. PMID- 14529504 TI - In silico ADME prediction: data, models, facts and myths. AB - A critical review of a very recent work in the field of in silico ADME prediction is presented with emphasis on the work published during the period 2000-2002, and several other review articles are mentioned in order to offer a broader view of the field. We find that not much progress has been made in developing robust and predictive models, and that the lack of accurate data, together with the use of questionable modeling end-points, has greatly hindered the real progress in defining generally applicable models. Due to the largely empirical nature of QSAR/QSPR approaches, general and truly predictive models for complex phenomena, such as absorption and clearance, may still be chimeric. The development of local models for use within focused chemical series may be the most appropriate way of utilizing in silico ADME predictions, once experience and data have been gained on a given project and/or structural class. PMID- 14529505 TI - Aspects of the stability and bioavailability of carbohydrates and carbohydrate derivatives. AB - Carbohydrates play a critical role in many biological processes and disease states including cancer, inflammation and infection. The development of carbohydrates as therapeutics continues to gain interest, as the biological roles of these biopolymers are further elucidated and understood. However, many carbohydrates display poor affinity, stability and bioavailability characteristics, which has led to a widely held view that this class of molecules make poor drugs. As there are already a significant number of carbohydrate-based drugs presently being employed by physicians, it is clear that some carbohydrates do make effective drugs. Recent advances in (a) the understanding of carbohydrate specific transport mechanisms, and (b) the development of novel carbohydrate based bioactives which may overcome many of the previous limitations of stability and bioavailability, suggest that carbohydrate-based compounds may provide a rich source of new drug candidates for a variety of diseases. PMID- 14529506 TI - Extending chemoselective ligation to sugar chemistry: convergent assembly of bioactive neoglycoconjugates. AB - Glycoproteins and glycolipids play central roles in human health and disease, and their mimetics are primary candidates for drug development. Our understanding of the molecular level of phenomena based on molecular recognition of oligosaccharides by specific receptors has taken tremendous advantage from their chemical synthesis, which provides homogeneous material not attainable from biosynthetic systems. This review summarizes chemoselective approaches for the assembly of neoglycoconjugates. The objective of these methods is to make glycoconjugate synthesis accessible to a broader community, thus accelerating progress in biotechnology. PMID- 14529507 TI - The potential of the molecular diversity of heparin and heparan sulfate for drug development. AB - Heparin and heparan sulfate have been shown to interact with a large number of biologically important proteins regulating important physiological processes. Specific oligosaccharide structures within the heterogeneous polysaccharide chains are responsible for the binding to individual proteins. Identification of specific protein-binding oligosaccharides provides lead compounds in pharmaceutical development and in one case has already resulted in an approved drug. The chemical and biosynthetic basis of the molecular diversity of heparin and heparan sulfate, its manifestation in heparin-protein interactions, and recent progress for drug development offered by this diversity are reviewed. PMID- 14529508 TI - Back to (non)-basics: recent developments in neutral and charge-balanced glycosidase inhibitors. AB - Certain glycosidase inhibitors possess potent antiviral, antitumour and antidiabetic properties. Glyconic acid lactones, the earliest glycosidase inhibitors identified, have planar anomeric carbons that mimic the transition state of glycoside hydrolysis. Other classes include lactams, glycals, epoxides, halides and sulfonium ions, the latter based on the natural product salacinol from an antidiabetic herb. PMID- 14529509 TI - Recent progress in the design of selectin inhibitors. AB - The selectins are a family of cell-adhesion proteins that mediate the early stages of leukocyte recruitment from the blood stream to sites of tissue damage through recognition of the carbohydrate epitope sialyl Lewis(x) (sLe(x)). Current development of small molecule based inhibitors of this process and their clinical potential to address numerous acute and chronic diseases are explored. PMID- 14529510 TI - Recent progress towards the identification of inhibitors of mycobacterial cell wall polysaccharide biosynthesis. AB - Mycobacterial infections have recently attracted significant attention from international health agencies due to the resurgence of these diseases worldwide. This review summarizes the recent work directed towards the identification of new anti-tuberculosis agents that act by inhibiting mycobacterial cell wall polysaccharide biosynthesis. PMID- 14529511 TI - Conformation of glycopeptides. AB - The presence of carbohydrate side-chains in native glycoproteins alters a number of biochemical properties of the peptide backbone. One of the most frequently studied questions is the conformation-modifying effect of sugar incorporation into asparagine, serine and threonine residues. When N-glycosylation modifies the conformation, the resulting structures are more ordered than the peptide chain without sugar addition. For O-glycopeptides the final conformations can be either more ordered or less ordered. In any event, only the innermost carbohydrates make contact with the peptide backbone. Through-space structural changes are mostly found downstream of the O-glycosylation site. In the repeat unit of epithelial mucin-1 protein, clustering of the carbohydrates results in an easily observable stabilization of the poly-proline II helix. PMID- 14529512 TI - Recent trends in carbohydrate modeling. AB - The exploding activities in modeling of carbohydrates during the past few years is reviewed with emphasis on advances in improving force fields, coupling of NMR measurements with molecular dynamics simulations, direct calculation of thermodynamic properties, application of quantum chemical methods on a large scale, and web-access to modeling. PMID- 14529513 TI - Neuropathic pain: some clues for future drug treatments. AB - Neuropathic pain is still far from being adequately dealt with. Under this name, several clinical entities have been considered and most of them only share several painful ailments. At present, the available treatments can only alleviate the pain of roughly half of the patients, and their effectiveness is often limited by the appearance of the intolerable side effects. In this review, we will consider the pathophysiology of neuropathic pain to understand the basis of pharmacological treatments that are currently being investigated. Some examples of these drugs will also be considered. PMID- 14529514 TI - VR1 receptor modulators as potential drugs for neuropathic pain. AB - The involvement of VR(1) in the endogenous pain signalling has converted this receptor into a promising therapeutic target for the development of a new family of potent analgesics devoid of the shortcomings of other analgesics commonly used. The desensitisation induced after VR(1) activation points to the utility of VR(1) agonists for the treatment of various nociceptive disorders including mitigation of neuropathic pain, inhibition of neurogenic inflammation and suppression of urinary bladder hyperreflexia, whereas VR(1) antagonists have been described as valuable agents for the treatment of inflammatory hyperalgesia and pain. Structure of the main classes of VR(1) ligands developed to date, their molecular mechanisms of action and their promising utility for the management of diverse nociceptive alterations, specially neuropathic pain, are discussed in this review. PMID- 14529515 TI - Small molecules targeting the NMDA receptor complex as drugs for neuropathic pain. AB - Pain is a complex disease that usually remains poorly treated or undertreated, especially the neuropathic pain caused by injury to the peripheral or central nervous system. Antagonists of the NMDA receptor complex have emerged as potential drugs for pain management. A strong case is being raised for non competitive or uncompetitive antagonists with low-to-moderate affinity and fast on/offset kinetics as drugs with good therapeutic profiles, because of their reduced side effects. PMID- 14529516 TI - AMPA glutamate receptors and neuropathic pain. AB - Glutamate receptors are implicated in many actions in the central nervous system, as an excitatory amino acid, and one of the more relevant is its role in excitotoxicity. Apart from this, it also has a role as pro-nociceptive agent, so that antagonizing its actions could be of interest for developing new analgesic agents. Furthermore, between the analgesics agents, it is of outstanding interest the fact that there is no specific therapy against the neuropathic pain, and glutamate receptor subunits have elicited as new potential targets for this disturbance. PMID- 14529517 TI - Cannabinoids and neuropathic pain. AB - After a brief overview of the endocannabinoid system (CB receptors, and endocannabinoids) and of the cannabinergic ligands, some general issues related to cannabinoids and pain are commented. Finally, the most important findings regarding cannabinoids and neuropathic pain are discussed in detail. PMID- 14529518 TI - Antidepressants in chronic neuropathic pain. AB - This review presents available clinical studies and new insights into mechanisms of analgesic effect and possible new routes of administration of antidepressant drugs. Older TCAs continue to be superior treatments. We focused on recent findings on newer antidepressants as analgesics. Their use should be supported by further controlled trials. PMID- 14529520 TI - Stages of HIV replication and targets for therapeutic intervention. AB - The replication cycle of human immunodeficiency virus (HIV) is divided into an early and a late phase. Most of the steps of the cycle have been targeted in antiviral therapy, although the drugs currently available for clinical use are only effective against two replication enzymes of the virus, either against the reverse transcriptase or against the viral protease. The introduction of combination anti-retroviral therapy changed the prognosis of HIV infection. HIV related morbidity and mortality rates in patients with advanced HIV infection have significantly declined. However, there are severe limits of HAART. Current anti-retroviral therapy do not allow viral eradication, therefore long-term use of the drugs is required. As a consequence, resistance develops in a significant portion of patients. Furthermore, several adverse metabolic side effects have been observed associated with the therapy. Therefore new approaches are required to control or eradicate this deadly virus infection. PMID- 14529519 TI - Drugs from the sea: conotoxins as drug leads for neuropathic pain and other neurological conditions. AB - The oceans are a source of a large group of structurally unique natural products that are mainly found in invertebrates such as sponges, tunicates, bryozoans, and molluscs. It is interesting to note that the majority of marine compounds currently in clinical trials or under preclinical evaluation are produced by these species rather than as secondary metabolites by marine algae. Through the combined efforts of marine natural products chemists and pharmacologists a number of promising compounds have been identified that are either already at advanced stages of clinical trials such as the new anti-cancer drug marine alkaloid ecteinascidin 743, or have been selected as promising candidates for extended preclinical evaluation. This is the case for conotoxins, (Table 1) where a number of conopeptides are currently being developed as analgesics for the treatment of neuropathic pain. PMID- 14529521 TI - Current treatment of HIV infection. AB - The field of antiretroviral therapy has witnessed remarkable progress during the past 15 years. There are now over a dozen approved therapeutic agents for infection with the human immunodeficiency virus (HIV), a pathogen that once caused nearly uniformly fatal illness. These agents usually target two essential enzymes of the virus, the reverse transcriptase and the protease. The era of potent antiretroviral therapy, also termed highly active antiretroviral therapy, began in 1996 and has been marked by dramatic declines in morbidity and mortality due to HIV disease in the developed world. These advances have not been without their cost in terms of drug resistance and toxicity and unwanted effects. Concern about these negative effects has led to a more conservative approach to the timing of the initiation of therapy and to clinical trials of intermittent therapy in an attempt to decrease the total exposure to drugs over time. Immune based approaches such as therapeutic vaccination may someday permit viremia to be controlled in the absence of drugs. Antiviral potency has been the key to the initial success of drug regimens, as well as to the durability of their success, the restoration of immune function, the prevention of the emergence of resistance, and ultimately the prevention of disease progression. Future progress in antiretroviral therapy will bring more choices for physicians and patients and will make an already complex field both more challenging and more rewarding. PMID- 14529522 TI - Strategies in the design of prodrugs of anti-HIV agents. AB - Control of AIDS requires development of special therapeutic strategies in order to reduce the level of monocyte/macrophage HIV infection, to prevent spread of HIV within the monocyte/macrophage reservoir, to maintain a therapeutically effective drug concentration in sanctuaries such as the brain and to overcome the problem of cellular resistance mechanisms. A popular approach towards this end has been the development of prodrugs of anti-HIV drugs. This review covers the different strategies devised for the design of prodrugs of anti-HIV agents with emphasis on the recent findings in this field of research. Thus, prodrugs of nucleoside reverse transcriptase inhibitors (NRTIs) including, 5'-O carboxylic ester derivatives, 5'-O- monophosphate analogues, macromolecular derivatives, prodrugs of purine nucleosides, prodrugs of acyclic nucleosides, homo and hetero dinucleotides, prodrugs of non-classical nucleoside analogues, boranophosphate triesters of NRTIs, and prodrugs of protease inhibitors including acyl substituted prodrugs, prodrugs with increased water solubility, monophosphate prodrugs, and conjugates of HIV protease inhibitors with a reverse transcriptase inhibitor through spontaneously cleavable linkers, constitute the subject of this review. PMID- 14529523 TI - Nonnucleoside inhibitors of HIV-1 reverse transcriptase: from the biology of reverse transcription to molecular design. AB - Replication of human immunodeficiency virus 1 (HIV-1) uses a viral reverse transcriptase (RT) to convert its positive-strand RNA into double stranded DNA, which is then integrated into host genome. Reverse transcription is a complex event involving p66 and p51 RT subunits but also several viral proteins including Nef, Tat, Vif, IN, NCp7 and p55gag. Viral RNA itself forms a primer/template complex by association with a cellular tRNA(Lys3) which is already present in mature virions. A RT initiation complex (RTIC) is thus formed which may also involve cellular protein upon viral entry. X rays diffraction and NMR studies of free or inhibitor-bound RT have led to the recognition of RT 3D structure, and allowed a thorough understanding of the mode of action of classical competitive nucleoside RT inhibitors (NRTIs) and of the binding of allosteric, non NRTIs (NNRTIs) inhibitors. This also opened an access to computer-aided drug design and modeling. Current NNRTIs represent, in terms of chemical structures, a heterogeneous class of inhibitors currently undergoing extensive development. By contrast with NRTIs, they seem to block initiation steps of reverse transcription. Molecular dynamics, detailed analysis of their interaction with RT as well as the incidence, in the series, of cases of non classical biological behavior, as illustrated here for a new family of compounds, suggest mechanisms of action which are not understandable without considering the involvement of the RTIC as a whole. This opens the exciting perspective of developing new compounds based on this integrated knowledge. Key Words: Nonnucleoside reverse transcriptase inhibitors (NNRTIs); Reverse transcriptase initiation complex (RTIC); Human immunodeficiency virus (HIV); Non classical nonnucleoside reverse transcriptase inhibitors; Molecular modeling; Docking; QSAR; Natural endogenous reverse transcription (NERT). PMID- 14529524 TI - Natural products with anti-HIV activity from marine organisms. AB - In order to combat the human immunodeficiency virus (HIV), diverse strategies have been developed to research on compounds which can be developed as therapeutic agents. Screening of natural products derived from numerous species has afforded metabolites with significant antiviral activity against the HIV. The marine environment representing approximately half of the global biodiversity offers an enormous resource for novel compounds. Currently more than 150 natural products with promising levels of anti-HIV activity have been isolated following bioassay guided protocols from aqueous or organic extracts of marine organisms. Some of the most characteristic marinemetabolites that have exhibited significant anti-HIV activity on different biochemical assays designed for chemotherapeutic strategies are: Cyanovirin-N, a protein from a blue green alga; various sulfated polysaccharides extracted from seaweeds (i.e. Nothogenia fastigiata, Aghardhiella tenera); the peptides tachyplesin and polyphemusin, which are highly abundant in hemocyte debris of the horseshoe crabs Tachypleus tridentatus and Limulus polyphemus; sponge metabolites such as avarol, avarone, ilimaquinone and several phloroglucinols; and a number of metabolites from marine fungi such as equisetin, phomasetin and integric acid. Considering that number of unique metabolites that have been isolated from a small extent of the ocean's biological and chemical diversity, the oceans represent a virtually untapped resource for the discovery of novel bioactive compounds. PMID- 14529525 TI - Therapeutic HIV vaccines. AB - Antiretroviral therapy with potent combinations of drugs has been responsible for a significant decline in the occurrences of AIDS defining conditions and death in the developed world. However, therapy requires life-long use and is complicated by relatively high failure rates, significant toxicities, adherence difficulties and the development of resistance. The combination of these complications of therapy and the availability of this treatment to only 1 in 20 of the estimated 34 million people infected world wide has prompted us to reconsider the current strategies for achieving the goals of HIV therapy. A more rational approach to therapeutic interactions is needed, particularly with respect to therapy in the developing world, with the focus shifted towards maintaining relative viral control over the long term. One potential mechanism to attain viral control over the long term is the use of therapeutic vaccines. This chapter will review the scientific rationale for therapeutic HIV-1 vaccines and the vaccines that have been evaluated as a therapeutic to date including recombinant envelope glycoproteins, inactivated envelope depleted virus, regulatory proteins such as Tat, cytokines such as IFN-(alpha), DNA vaccines, and live viral vectors. Although the future role of therapeutic vaccines in the treatment of HIV-1 remains to be determined, at a minimum this immunomodulatory approach will provide new insights into fundamental viral-host cell interactions and the pathogenesis of HIV-1. Yet even more notable, is that, if successful, a therapeutic vaccine product would be inexpensive and rapidly exportable representing a treatment strategy for people living with HIV infection worldwide. PMID- 14529526 TI - Functions of propeptide parts in cysteine proteases. AB - Regulation of proteolytic enzyme activity is an essential requirement for cells and tissues because proteolysis at the wrong time and location may be lethal. Two principal mechanisms to control the activity of proteases have been developed during evolution. The first is the co-evolution of endogenous inhibitors, typically occurring in cellular compartments separated from those containing active enzymes. The second is the fact that proteases are synthesized as inactive or less active precursor molecules. They are activated, in some cases, upon an appropriate signal like acidification, Ca(++) -binding or, in other cases, by limited intra- or intermolecular proteolysis cleaving off an inhibitory peptide. These regulatory proenzyme regions have attracted much attention during the last decade, since it became obvious that they harbour much more information than just triggering activation. In this review we summarize experimental data concerning three functions of propeptides of clan CA family C1 cysteine peptidases (papain family), namely the selectivity of their inhibitory potency, the participation in correct intracellular targeting and assistance in folding of the mature enzyme. Cysteine peptidases of the CA-C1 family include members from the plant kingdom like papain as well as from the animal kingdom like the lysosomal cathepsins L and B. As it will be shown, the functions are determined by certain structural motifs conserved over millions of years after the evolutionary trails have diverged. The function of propeptides of two other important classes of cysteine peptidases - the calpains, clan CA family C4, and the caspases, clan CD family C 14 - are not considered in this review. PMID- 14529527 TI - Crystallographic and bioinformatic studies on restriction endonucleases: inference of evolutionary relationships in the "midnight zone" of homology. AB - Type II restriction endonucleases (ENases) cleave DNA with remarkable sequence specificity. Their discovery in 1970 and studies on molecular genetics and biochemistry carried out over the past four decades laid foundations for recombinant DNA techniques. Today, restriction enzymes are indispensable tools in molecular biology and molecular medicine and a paradigm for proteins that specifically interact with DNA as well as a challenging target for protein engineering. The sequence-structure-function relationships for these proteins are therefore of central interest in biotechnology. However, among numerous ENase sequences, only a few exhibit statistically significant similarity in pairwise comparisons, which was initially interpreted as evidence for the lack of common origin. Nevertheless, X-ray crystallographic studies of seemingly dissimilar type II ENases demonstrated that they share a common structural core and metal binding/catalytic site, arguing for extreme divergence rather than independent evolution. A similar nuclease domain has been also identified in various enzymes implicated in DNA repair and recombination. Ironically, following the series of crystallographic studies suggesting homology of all type II ENases, bioinformatic studies provided evidence that some restriction enzymes are in fact diverged members of unrelated nuclease superfamilies: Nuc, HNH and GIY-YIG. Hence, the restriction enzymes as a whole, represent a group of functionally similar proteins, which evolved on multiple occasions and subsequently diverged into the "midnight zone" of homology, where common origins within particular groups can be inferred only from structure-guided comparisons. The structure-guided approaches used for this purpose include: identification of functionally important residues using superposition of atomic coordinates, alignment of sequence profiles enhanced by secondary structures, fold recognition, and homology modeling. This review covers recent results of comparative analyses of restriction enzymes from the four currently known superfamilies of nucleases with distinct folds, using crystallographic and bioinformatic methods, with the emphasis on theoretical predictions and their experimental validation by site-directed mutagenesis and biochemical analyses of the mutants. PMID- 14529528 TI - The design, synthesis and application of stereochemical and directional peptide isomers: a critical review. AB - Physiological processes are regulated to a large extent by physical and chemical interactions between polypeptides. Although many small molecules have been discovered that can modulate such interactions and may be useful as drugs, the design of these agents purely from the knowledge of the details of a given protein-protein interaction, or through screening, remains difficult. Therefore, the peptidomimetic process, which aims at using peptides derived from either polypeptide binding partner directly, or after modification to improve affinity and physicochemical properties, continues to be attractive. The vast majority of naturally occurring polypeptides are composed of L-amino acids. Because natural proteins need to be metabolised, L-amino acid polypeptides are very prone to proteolytic degradation, a property that severely limits their therapeutic application. The proteolytic machinery is not well equipped to deal with D-amino acid polypeptides, however, and it is this finding above all else that has spurned research into stereochemical and directional manipulation of peptide chains. The expectation has been that systematic inversion of the stereochemistry at the peptide backbone alpha-carbon atoms, if accompanied by chain reversal, should yield proteolytically stable retro-inverso peptide isomers, whose side chain topology, in the extended conformation, corresponds closely to that of a native sequence, and whose biological activity emulates that of a parent polypeptide. The actual structural implications of modifying amino acid stereochemistry and peptide bond direction are reviewed critically here and the reasons for the lack of general success with this strategy are discussed. The application of polypeptides is particularly pertinent to synthetic vaccine design. Interestingly, the retro-inverso strategy has been more successful for immunological applications than elsewhere; recent finding are collated in this review. Partial rather than global retro-inversion holds much promise since the loss of crucial backbone hydrogen-bonding through peptide bond reversal can be avoided, while still permitting stabilisation of selected hydrolysis-prone peptide bonds. Generically applicable synthetic methods for such partially modified retro-inverso peptides are not as yet available; progress towards this goal is also summarised. PMID- 14529529 TI - Is use of the hydrophobic moment a sound basis for predicting the structure function relationships of membrane interactive alpha-helices? AB - Amphiphilic alpha-helices play a fundamental role in protein membrane association and show a segregation of polar and apolar amino acid residues. Based on correlations between amphiphilic properties and biological function, a number of theoretical approaches have been developed, which quantify alpha-helix amphiphilicity and then attempt to assign function. The most commonly used measure of amphiphilicity is the hydrophobic moment, < microH >, which, when used in conjunction with an alpha-helix's mean hydrophobicity, < H >, has been used to classify membrane interactive amphiphilic alpha-helices as either surface active or transmembrane. Here, the predictive efficacy of plot methodology is reviewed by examining published data, which compare the function of known membrane interactive amphiphilic alpha-helices to that assigned by this methodology. The results of this review are discussed in relation to the reliability of < microH > as a quantifier of alpha-helical amphiphilicity, and the ability of < microH > and < H > to describe alpha-helical structure / function relationships. It is concluded that hydrophobic moment plot methodology is not a generally reliable predictor of alpha-helical structure / function relationships. It appears that the inefficacy of plot methodology is primarily due to the inability of the plot diagram to accommodate the heterogeneity of the alpha-helical classes it attempts to define. However, the predictive efficacy of the methodology appears to be improved if other alpha-helical parameters are also considered when assigning alpha-helical function. It is suggested that the conventional methodology should be seen only as an indicator for the assignation of structure / function relationships, providing a guide to future experimental investigations. PMID- 14529530 TI - Structural biology of the cell adhesion protein CD2: from molecular recognition to protein folding and design. AB - CD2 (cluster of differentiation 2) is a cell adhesion molecule expressed on T cells and is recognized as a target for CD48 (rats) and CD58 (humans). Tremendous progress has been achieved in understanding the function of CD2, the mechanism of molecular recognition and protein folding, thus, leading towards the use of this protein as a scaffold for protein design. CD2 has been shown to set quantitative thresholds in T cell activation both in vivo and in vitro. Further, intracellular CD2 signaling pathways and networks are being discovered by the identification of several cytosolic tail binding proteins. In addition, a new method for directly measuring heterophilic adhesion has been developed. The functional "hot spot" for the adhesion surface of CD2 and CD58 has been dissected. Detailed NMR studies reveal that rat CD2 weakly self-associates to form a homodimeric structure in solution. Dynamic interaction of CD2 with the GYF and SH3 domains has been investigated. CD2 has been shown to form fibrils in the presence of 2,2,2 trifluoroethanol (TFE) and at low pH. Furthermore, kinetic studies have been completed to monitor the effect of surface hydrophobic residues and intramolecular bridges on the folding pathways of CD2. Our lab has de novo designed single calcium-binding sites into domain 1 of rat CD2 (CD2-D1) with strong metal selectivity. In addition, the EF-hand motifs have been grafted into CD2 to understand the site-specific calcium-binding affinity of calmodulin and calcium-dependent cell adhesion. PMID- 14529531 TI - Monitoring intracellular proteins using fluorescence techniques: from protein synthesis and localization to activity. AB - The recent breakthroughs in genomics and proteomics and improvements of optical methods have made it possible to obtain localized, real-time information on intracellular proteins dynamics, through dynamic three-dimensional (3D) maps of the living cell with nanometric resolution of individual molecules. On one side, brighter variants of the Green Fluorescence Protein (GFP) have been engineered that have different excitation and/or emission spectra that better match available light sources. Like their parent molecule, these variants retain their fluorescence when fused to heterologous proteins on the N- and C- terminals, and this binding generally does not affect the functionality of the tagged protein leading the way to their use as an intracellular reporter. On the other side, optical methods have been improved to allow reaching the level of single-molecule detection inside living cells. Nevertheless some limitations exist for the use of GFP variants for probing 3D conformational changes of proteins. First, these variants are fused to the N and/or C terminals of the studied protein, which are generally not the best location to detect conformational changes resulting from the binding to other proteins or enzyme substrates. Then their own relatively large size makes them unusable for tagging small proteins. These limitations suggest that new tagging processes, permitting the location of the right fluorescent markers at the right places, must be found to built up inter- and/or intra-molecular rulers allowing one to monitor conformational changes resulting from intracellular protein-protein, protein-membrane, and enzyme-substrate binding. These specific locations can be obtained from in vitro studies of 3D conformational changes that occur during protein docking. PMID- 14529532 TI - Design of synthetic peptides for diagnostics. AB - Due to the advantageous properties of synthetic molecules compared to biological ones biological molecules in diagnostic tests are replaced increasingly by synthetic ones, usually synthetic peptides or related molecules. The replacement of biological antigens by synthetic peptides is most advanced at present, as well as the use of site-specific antibodies induced with synthetic peptides. Moreover recent results indicate that synthetic molecules may also replace antibodies. Ultimately this will lead to diagnostic assays built of synthetic molecules only. PMID- 14529533 TI - Synthetic peptides in the diagnosis of systemic autoimmune diseases. AB - Proteins recognized by antibodies from patients with autoimmune diseases have been intensively studied over the two past decades since cDNAs encoding autoantigens have become available. Identity of many of them has been defined, and specific structural motifs or post-translational modifications, which may be important to explain the generation of such antibodies during the autoimmune process, have been pointed out. Immunological analysis of sera from autoimmune patients with recombinant fragments and with short peptides has revealed the presence of dominant epitopes along proteins; some of them are targeted by antibodies from patients with specific diseases or disease subsets. Innovative technologies such as peptide arrays and biosensors as well as the exploitation of large peptides libraries have recently open up new perspectives. Peptides bearing natural modifications, peptide analogues, as well as mimotopes of protein or non protein antigens (DNA, RNA, sugar) have been developed and might advantageously replace native antigens in routine immunoassays. Although numerous conformational epitopes have not yet been identified, and cannot be identified by the approaches classically used in epitope mapping studies, such peptides and peptide analogues may represent efficient probes to detect the presence of circulating autoantibodies in the serum of autoimmune patients and help for establishing specific and sensitive early diagnostic tests. They may also lead to the design of high-affinity ligands for purifying autoantibodies. These different aspects are discussed and epitope mapping studies of a number of autoantigens (e.g. histones, sn and hnRNP proteins and Ro proteins) are summarized. PMID- 14529534 TI - Synthetic peptides in the diagnosis of neurological diseases. AB - For several years peptides have been used to investigate many aspects of the molecular mechanisms supporting the etiopathogenesis of neurological diseases. Even if well-established and commercially-available assays for the diagnosis of neurological diseases, based on peptides, are presently not available, much research has been focused to the comprehension of the pathogenetic mechanisms of relevant neurological diseases, such as Multiple Sclerosis, Alzheimer's disease and prion diseases. Several peptides, which strongly contributed to increase in the progress, and in the understanding of the molecular mechanisms of these important diseases, have been selected as antigens to be proposed in possible alternative diagnostic methodologies. The authors focused their attention on the possible peptides to be used as diagnostics for these three characteristic neurological diseases. In fact, Multiple Sclerosis is an autoimmune disease in which peptides have been used for the characterization of both B and T immune response, while Alzheimer's disease and prion diseases can be cumulative regarded as "protein folding disorders" in which peptides can be used to identify the misfolded protein. PMID- 14529535 TI - Synthetic peptides in the diagnosis of HIV infection. AB - Peptide-based enzyme-linked immunosorbent assays have been found to be enough sensitive and specific for the diagnosis of human immunodeficiency virus specific antibodies in acquired immunodeficiency syndrome patients. This review provides an overview of the most important peptides developed for use as synthetic antigens in immunodiagnosis of HIV-infected patients. In particular, many studies have been devoted to discriminate between the two retroviruses HIV-1 and HIV-2, as well as different subtypes. PMID- 14529536 TI - GB virus C (GBV-C) / hepatitis G virus (HGV): towards the design of synthetic peptides-based biosensors for immunodiagnosis of GBV-C/HGV infection. AB - In the present study, new putative epitopes located in structural (E2) and non structural (NS3) proteins of GBV-C/HGV were identified by computer-aided prediction of antigenicity and synthesized in solid-phase, following an Fmoc/tBut strategy, for their use in immunoassays. The corresponding synthetic peptides were used as antigens in ELISA assays and in real-time biospecific interaction measurements. This last approach allowed direct detection of GBV-C/HGV-specific antibodies in human sera. Good correlations were obtained between the biospecific interaction analysis and the ELISA. To verify the performance of these new assays in comparison to the existing recombinant E2 protein commercial test, antibodies to synthetic peptides were searched for in different panels of serum samples. The main conclusion of this work is the usefulness of E2 peptides in the detection of antibodies. Moreover, the NS3 peptide could be exploited to improve the sensitivity of the currently available test. Our results offer a new approach to develop new diagnostic peptide based biosensors for serodiagnosis of GBV-C/HGV infection. PMID- 14529537 TI - Immunodiagnosis of parasitic diseases with synthetic peptides. AB - Parasitic diseases remain as a major public health problem worldwide, not only based on their historically high morbidity and mortality rates, but also because risk factors associated with their transmission are increasing. Laboratory diagnosis and particularly immunodiagnosis is a basic tool for the demonstration, clinical management and control of these infections. Classically, the serological tests for the detection of antibodies or antigens are based on the use of crude and purified antigens. Synthetic peptides have opened a new field and perspectives, as the source of pure epitopes and molecules for diagnosis of malaria, Chagas' disease, leishmaniasis, schistosomiasis, hidatidosis, cysticercosis and fasciolosis based on the detection of antibodies and circulating antigens. Herein, are critically reviewed the relevant advances and applications of the synthetic peptides on immunodiagnosis of parasitic diseases. A variety of sequences, constructs (monomers, polymers, MAPs), immunological methods and samples have been used, demonstrating their diagnostic potential. However, in most parasitic infections it is necessary to use more than a single peptide in order to avoid the genetic restriction against certain epitopes, as well as to test them in well characteized groups of patients, in order to confirm their sensitivity and specificity. The concept of multidiagnosis with synthetic peptides, using a novel multi-dot blot assay is introduced. Finally, the chemical imitation of antigens, offers a tremendous posibilities in the diagnosis of parasitic infections in developing countries since this strategy is cheaper, simpler, reproducible, useful for large scale testing and in most cases, specific and sensitive. PMID- 14529538 TI - New insights into the molecular mechanisms of action of bisphosphonates. AB - Bisphosphonates are currently the most important and effective class of anti resorptive drugs available, but the exact molecular mechanisms by which they inhibit osteoclast-mediated bone resorption have only recently been identified. Due to the targeting of bisphosphonates to bone mineral and the ability of osteoclasts to release bone-bound bisphosphonate, a direct effect on mature osteoclasts appears to be the most important route of action. As a result of recent discoveries concerning their molecular mechanism of action, bisphosphonates can be grouped into two classes. The simple bisphosphonates that closely resemble PPi (such as clodronate, etidronate and tiludronate) can be metabolically incorporated into non-hydrolysable analogues of ATP that accumulate intracellularly in osteoclasts, resulting in induction of osteoclast apoptosis. By contrast, the more potent, nitrogen-containing bisphosphonates (such as pamidronate, alendronate, risedronate, ibandronate and zoledronate) appear to act as analogues of isoprenoid diphosphate lipids, thereby inhibiting FPP synthase, an enzyme in the mevalonate pathway. Inhibition of this enzyme in osteoclasts prevents the biosynthesis of isoprenoid lipids (FPP and GGPP) that are essential for the post-translational farnesylation and geranylgeranylation of small GTPase signalling proteins. Loss of bone-resorptive activity and osteoclast apoptosis is due primarily to loss of geranylgeranylated small GTPases. Identification of FPP synthase as the target of nitrogen-containing bisphosphonates has also helped explain the molecular basis for the adverse effects of these agents in the GI tract and on the immune system. PMID- 14529539 TI - Calcitonin for osteoporosis and bone pain. AB - Calcitonin has been approved for the treatment of osteoporosis and other diseases involving accelerated bone turnover for approximately 25 years. The most commonly studied and prescribed form is salmon calcitonin, which has a greater efficacy in clinical use. A wealth of well-controlled clinical studies have demonstrated that calcitonin preserves or increases bone mineral density (BMD) and reduces the risk of vertebral fractures in osteoporosis. Recent studies have indicated that while a low BMD is correlated with an increase in fracture risk, increases in BMD alone do not explain the antifracture efficacy of antiresorptive therapies such as calcitonin. Therapies that moderately increase BMD may reduce fracture risk by reducing the rate of bone turnover and maintaining the integrity of the trabecular architecture, resulting in the preservation of bone strength and quality in osteoporotic patients. An advantage of calcitonin that is not shared by other antiresorptive therapies is its direct analgesic effect on bone pain. Calcitonin has been demonstrated to be clinically useful in improving pain, not only from the acute vertebral fractures of osteoporosis, but also in Paget's disease, bone malignancies, and other sources of musculoskeletal pain. Drugs containing calcitonin may be approved for additional indications in the near future, and as more convenient routes of administration such as the oral route become available, the demand for the calcitonin peptide is expected to increase. PMID- 14529541 TI - Dietary intake and bone status with aging. AB - Osteoporosis and related fractures represent major public health problems that are expected to increase dramatically in importance as the population ages. Dietary risk factors are particularly important, as they are modifiable. However, most of the attention to dietary risk factors for osteoporosis has focused almost exclusively on calcium and vitamin D. Recently, there has been considerable interest in the effects of a variety of other nutrients on bone status. These include minerals--magnesium, potassium, copper, zinc, silicon, sodium; vitamins- vitamin C, vitamin K, vitamin B12, vitamin A; and macronutrients--protein, fatty acids, sugars. In addition, foods and food components, including milk, fruit and vegetables, soy products, carbonated beverages, mineral water, dietary fiber, alcohol and caffeine have recently been examined. Together the evidence clearly suggests that prevention of bone loss through diet is complex and involves many nutrients and other food constituents. For many, results remain inconclusive and in some cases contradictory. However, it is increasingly clear that our exposure to a complex of nutrients and food constituents interacts to affect bone status. In addition to identifying the role of individual components, there is a great need to understand the interactions of these factors within diets and, increasingly, in the presence of nutrient supplements. Furthermore, genetic factors are likely to interact with these dietary exposures, increasing the complexity of these effects. With advances in both genetics and nutrition, improved understanding of all these interactions will contribute to effective recommendations for prevention of bone loss and osteoporosis in the aging population. PMID- 14529540 TI - Runx2/Cbfa1: a multifunctional regulator of bone formation. AB - Runx2/Cbfa/AML3 is a member of the runt homology domain family of transcription factors, essential for osteoblast differentiation and bone formation. Defining the molecular mechanisms by which Runx2 can function as a master regulatory gene for activating the program of osteoblastogenesis has provided novel insights for transcriptional regulation of tissue-specific genes. Regulation of Runx expression has the potential to serve as a basis for the design of novel therapeutic strategies for promoting bone formation. Here we review the unique properties of Runx2 that mediate several key functions necessary for regulating skeletogenesis, controlling osteoblast growth and differentiation, and integrating the complex pathways required for bone formation and turnover. PMID- 14529542 TI - Novel inhibitors of HIV integrase: the discovery of potential anti-HIV therapeutic agents. AB - The viral enzyme, HIV integrase (MW 32 kDa), is one of the three key enzymes of the pol gene of HIV. HIV integrase is involved in the integration of HIV DNA into host chromosomal DNA. There is apparently no functional equivalent of this enzyme in human cells. Integration of HIV DNA into the host cell genome apparently occurs by a carefully defined sequence of DNA tailoring (3'-processing) and coupling (joining or integration) reactions. In spite of some effort in this area targeted at the discovery of therapeutically useful inhibitors of this viral enzyme, there are no drugs for HIV/AIDS in clinical use where the mechanism of action is inhibition of HIV integrase. It is clear that new knowledge on inhibitors of this enzyme is of critical importance in the anti-HIV drug discovery area. This review focuses on the major classes of compounds that have been discovered as inhibitors of HIV integrase. Some of these compounds are non specific inhibitors of the enzyme while evidence suggests that others may possess some specificity. The various classes include nucleotides, oligonucleotides, dinucleotides, and miscellaneous small molecules including heterocyclic systems, natural products, diketo acids and sulfones. A major focus of the review is on discoveries from my laboratory in the area of non-natural, nuclease-resistant dinucleotide inhibitors of HIV integrase. PMID- 14529543 TI - Phosphonomethoxyalkyl analogs of nucleotides. AB - Acyclic nucleoside phosphonates (ANP) are analogs of nucleotides exhibiting various biological (e.g. antiviral, cytostatic, antiparasitic, immunomodulatory) activities. This review describes synthetic approaches to their preparation, biological activities, structure-activity relationships and mode of action of ANPs, with emphasis to selected drugs or drug candidates. Three compounds of the ANP group (cidofovir, adefovir, tenofovir) are active components of potent antivirals approved for therapeutic use in human medicine aimed at hepatitis B, AIDS and various diseases caused by DNA viruses. PMID- 14529544 TI - Anticancer antifolates: current status and future directions. AB - Antifolates are the oldest of the antimetabolite class of anticancer agents and were one of the first modern anticancer drugs. The first clinically useful antifolate, described in 1947, was 2,4-diamino-pteroylglutamate (4-amino-folic acid; aminopterin; AMT) which yielded the first-ever remissions in childhood leukemia. AMT was soon superseded by its 10-methyl congener, methotrexate (MTX), based on toxicity considerations; MTX remains, with one limited exception, the only antifolate anticancer agent in clinical use to this date. Because of the safety and utility of MTX, considerable effort has been invested in attempting to design more therapeutically selective antifolates or antifolates with a wider tumor spectrum. Initially, the design was based on the burgeoning knowledge of folate-dependent pathways and the determinants of the mechanism of action of MTX. These determinants include transport, the tight-binding inhibition of its target (the folate-dependent enzyme dihydrofolate reductase (DHFR)), and metabolism of MTX to poly-gamma-glutamate (Glu(n)) metabolites. These early studies led to the development of other antifolate DHFR inhibitors of two types: (1). "classical" analogs that use the same cellular transport systems as MTX and are also metabolized to Glu(n); and (2). "nonclassical" (i.e., lipophilic) analogs that do not require transport systems and that are not metabolized to Glu(n). Although several of these analogs have undergone clinical trial, none is proved superior to MTX. Detailed examination of the mechanisms of cytotoxicity and selectivity of MTX showed that inhibition of both dTMP synthesis and de novo purine synthesis, secondary to DHFR inhibition, led to DNA synthesis inhibition and subsequent cell death; inhibition of other folate-dependent pathways did not appear necessary for cell death. Further studies showed that the contribution of inhibition of dTMP or purine synthesis to cell death varied in different cell types. These data suggested that inhibition of one of these pathways individually might (at least in some cases) be therapeutically superior to the dual inhibition induced by MTX. Thus in rational design and in structure-based design studies, two new classes of antifolate enzyme inhibitors were elaborated-direct inhibitors of thymidylate synthase (TMPS) and direct inhibitors of one or both of the two folate-dependent enzymes of de novo purine synthesis. Members of each class included both classical and nonclassical types. After preclinical evaluation, several of these have moved into clinical trials. To date only one new TMPS inhibitor has successfully completed clinical trials and been approved for routine use; this drug, Tomudex (D1694, raltitrexed) is currently approved only in Europe and only for the treatment of colon cancer. This still represents a step forward for antifolates, however, since MTX is well-known to be ineffective in colon cancer; thus Tomudex extends the tumor range of antifolates. Antifolate development continues. Based on the immense body of knowledge now extant on antifolates, specific aspects of the mechanism of action have been the focus. Newer antifolates have been described that inhibit more than one pathway in folate metabolism, that have improved delivery, or that inhibit other targets in folate metabolism. These new analogs are in various stages of preclinical and clinical development. PMID- 14529545 TI - Deaza analogs of folic acid as antitumor agents. AB - Derivatives of the vitamin folic acid function in the body for the synthesis of thymidylate, purines and amino acids and are necessary for normal metabolism and growth. Methotrexate (MTX), an inhibitor of dihydrofolate reductase (DHFR) is the outstanding example of an antitumor antifolate. MTX is clinically useful in the treatment of childhood leukemia, choriocarcinoma and psoriasis, where it corrects abnormal growth, and in rheumatoid arthritis and other autoimmune diseases where it corrects abnormal immune function. Since 1949, when the chemical synthesis of MTX was reported by workers at the Lederle Laboratories of the American Cyanamid Company, much has been learned about the basis of antifolate cytotoxicity and selectivity. This review will focus on deaza antifolates which are: 1). presently under clinical development and 2). less developed compounds which represent novel approaches. Compounds will be grouped according to their enzyme targets; DHFR, thymidylate synthase (TS) and glycinamide ribonucleotide formyltransferase (GARFT). In addition to inhibition of target enzymes, antifolate membrane transport into cells and conversion to poly-L-gamma-glutamate forms are important considerations in drug design along with the reverse processes, cellular hydrolysis of antifolate poly-L-gamma-glutamates to monoglutamates and the extrusion of the monoglutamates through the cell membrane. These processes can be modulated by competition with folates. PMID- 14529546 TI - Thioguanine, mercaptopurine: their analogs and nucleosides as antimetabolites. AB - 6-Mercaptopurine (6MP) and 6-thioguanine (6TG) are analogs of the natural purines: hypoxanthine and guanine. Both mercaptopurine and thioguanine are substrates for hypoxanthine-guanine phosphoribosyltransferase and are converted into the ribonucleotides 6-thioguanosine monophosphate (6-thioGMP) and 6 thioinosine monophosphate (T-IMP) respectively. The accumulation of these monophosphates inhibits several vital metabolic reactions. Today, these thiopurine bases remain valuable agents for the induction and maintenance of remissions in patients with myelocytic and acute lymphocytic leukemia. Despite their proved clinical importance, 6MP and 6TG have certain therapeutic disadvantages, which have continued to stimulate the search for purine derivatives enhancing therapeutic efficacy. Considerable efforts have been made to prepare other novel mercaptopurine and thioguanine analogs and their nucleosides to improve the antitumor efficacy. The effectiveness of these thiopurines against certain tumor cell lines suggested that some of these mercaptopurine analogs and their nucleosides would be worthy of consideration in order to determine whether they exert a more selective effect against neoplastic cells than against normal cells or they might be useful in patients whose disease has become resistant to 6MP or 6TG. This review will focus on mercaptopurine analogs and their nucleosides as antimetabolite agents. PMID- 14529547 TI - Role of heme oxygenase-1 in cardiovascular function. AB - Heme oxygenase (HO) is a cytoprotective enzyme that degrades heme (a potent oxidant) to generate carbon monoxide (a vasodilatory gas that has anti inflammatory properties), bilirubin (an antioxidant derived from biliverdin), and iron (sequestered by ferritin). Due to the properties of inducible HO (HO-1) and its products, we hypothesized that HO-1 would play an important role in the regulation of cardiovascular function. In this article we will review the role of HO-1 in cardiovascular function, and highlight our previous studies using gene deletion and gene overexpression transgenic approaches in mice. These studies will include the investigation of HO-1 in the setting of hypertension (renovascular), atherosclerosis and vascular injury (vein graft stenosis), hypotension (endotoxemia), and ischemia/reperfusion injury (heart). In a chronic renovascular hypertension model, blood pressure elevation, cardiac hypertrophy, acute renal failure, and acute mortality induced by one kidney-one clip surgery are more severe in HO-1 null mice. Moreover, absence of HO-1 leads to accelerated atherosclerotic lesion formation and vein graft disease. In addition, HO-1 null mice with endotoxemia have earlier resolution of hypotension, yet the mortality and the incidence of end organ damage are higher in the absence of HO-1. In contrast, mice with cardiac-specific overexpression of HO-1 have an improvement in cardiac function, smaller myocardial infarcts, and reduced inflammatory and oxidative damage after coronary artery ligation and reperfusion. Taken together, these studies suggest that an absence of HO-1 has detrimental consequences, while overexpression of HO-1 plays a protective role in ischemia/reperfusion injury. PMID- 14529548 TI - Heme oxygenase-1 against vascular insufficiency: roles of atherosclerotic disorders. AB - Heme oxygenase (HO), an enzyme essential for heme degradation, shows anti oxidative and anti-inflammatory properties via the production of bile pigments, carbon monoxide (CO) and ferritin induction under various pathophysiological conditions. A number of recent studies have shown biological effects of HO reaction in cardiovascular disorders. An inducible form of HO, HO-1, is induced by a variety of stresses such as oxidized lipoproteins, cytokines, hemodynamic changes, angiotensin II and nitric oxide (NO) in vascular wall. HO-1 induction seems to function as an adaptive response against these injurious stimuli. HO-1 induction in artery wall scavenges reactive oxygen species, which leads to the attenuation of monocyte adhesion and chemotaxis. HO-1 induction also reduces lipid peroxidation in plasma and artery wall. These properties of HO-1 suggest anti-atherogenic roles of this enzyme. In this review, roles of endothelial HO-1 expression and bilirubin in atherogenesis are also discussed. HO-1 also seems to play a significant role in restenosis after angioplasty, which is a major clinical problem associated with atherosclerosis. Recent progress in human HO-1 genetics supports these experimental results. This review aims to reaffirm current problems in the biological aspects of HO and suggest future research direction and clinical application. PMID- 14529549 TI - Transcriptional regulation of the heme oxygenase-1 gene via the stress response element pathway. AB - The heme oxygenase-1 (HO-1) enzyme catalyzes the rate-limiting reaction in the catabolism of heme yielding products with pleiotropic, but ultimately, cytoprotective activities. High levels of HO-1 are frequently detected in various pathological states and generally in states of cellular oxidative stress. Induction of HO-1, regulated at the level of gene transcription, is essential for manifestation of the enzyme's cytoprotective function. Extensive analysis of the mouse gene, and to a lesser extent of the human gene, has identified a common mechanism the stress response element (StRE)/Nrf2 transcription factor pathway for gene regulation in response to a diverse array of HO-1 inducers including the substrate heme, various environmental and industrial toxins, and plant-derived polyphenolic compounds. In addition to Nrf2 complexes, numerous dimeric transcription factors bind to the StRE, permitting induction, repression and overall fine-tuning of gene activity. In principle, the multiplicity of StRE binding proteins also provides for a range of pharmaceutical targets for controlled production of the potentially therapeutic HO-1 protein. PMID- 14529550 TI - Therapeutic applications of human heme oxygenase gene transfer and gene therapy. AB - The search for methods to control physiological levels of carbon monoxide (CO), a vasoactive molecule, and bilibrubin, an antioxidant, have improved our understanding of the protective role of heme oxygenase (HO) against oxidative injury. HO activity can assist other antioxidant systems in diminishing the overall production of reactive oxygen species, thus contributing to cellular resistance to such injury. Overexpression of HO gene by targeted gene transfer has become a powerful tool for studying the role of this human enzyme. Successful and functional HO gene transfer requires two essential elements. First, the HO gene must be delivered into a safe vector, such as the adenoviral, retroviral and leptosome based vectors that are currently being used in clinical trials. The use of non-viral vectors has also been described. Second, with the exception of HO gene delivery to ocular or cardiovascular tissue via catheter-based interventions, HO gene delivery must be site and organ specific. Site-specific delivery of HO-1 to renal structures in SHR, specifically mTAL, using Na+-K+ Cl- cotransporter (NKCC2 promoter), has been shown to normalize blood pressure and provide protection to mTAL against oxidative injury, respectively. Human HO-1 gene transfer into endothelial cells has been shown to attenuate Ang II- TNF- and heme-mediated DNA damage. Furthermore, delivery of human HO-1 into SHR has been shown to enhance somatic body growth and cell proliferation. The ability to transfect human HO gene and to demonstrate its expression may offer a new therapeutic strategy for treating pathological conditions, such as hypertension, trauma and hemorrhage. PMID- 14529551 TI - Bioactivity and pharmacological actions of carbon monoxide-releasing molecules. AB - Carbon monoxide (CO) is a resourceful gas as recent advances in the area of cell signaling are revealing an unexpected physiological role for CO in the cardiovascular, immune and nervous systems. Transition metal carbonyls have been lately discovered to function as CO-releasing molecules (CO-RMs) and elicit distinct pharmacological activities in biological systems. Studies currently ongoing in our laboratories are investigating both the chemical and bioactive features of a series of water-soluble CO-RMs and their specific utilization as vasoactive mediators, anti-inflammatory agents and inhibitors of cellular and tissue damage. The data presented in this review corroborate the notion that transition metal carbonyls could be used as carriers to deliver CO in mammals and highlight the bioactivity and potential therapeutic features of CO-RMs in the mitigation of cellular and organ dysfunction. PMID- 14529552 TI - Anti-inflammatory actions of the heme oxygenase-1 pathway. AB - Heme oxygenase 1 (HO-1) is induced by oxidative or nitrosative stress, cytokines and other mediators produced during inflammatory processes, likely as part of a defence system in cells exposed to stress to provide a negative feedback for cell activation and the production of mediators, which could modulate the inflammatory response. HO-1 activity results in the inhibition of oxidative damage and apoptosis, with significant reductions in inflammatory events including edema, leukocyte adhesion and migration, and production of inflammatory cytokines. HO-1 is induced by nitric oxide (NO) in different biological systems and can control the increased production of this mediator observed in many inflammatory situations. Regulatory interactions between HO-1 and cyclooxygenase (COX) pathways have also been reported. Modulation of signal transduction pathways by HO-1 or products derived from its activity, such as carbon monoxide (CO), may mediate the anti-inflammatory effects of this protein. Regulation of HO-1 activity may be a therapeutical strategy for a number of inflammatory conditions. PMID- 14529553 TI - Control of blood pressure in the community: an unsolved problem. AB - Available data indicate that blood pressure (BP) is reduced below 140-90 mmHg in less than 30% of hypertensive patients. This poor control of BP derives from lack of diagnosis (unawareness of hypertension), lack of treatment of aware hypertensive patients and lack of efficacy of treatment. Systolic BP (SBP) is now considered as the most important parameter for diagnosis and stadiation of hypertension, above all in elderly patients, and the most frequent cause of unsatisfactory control of BP in the population. Lack of SBP control is caused both by physicians' attitude and difficulty in reducing SBP. Physicians are more prone to consider diastolic BP as the most important parameter for diagnosis and stadiation of hypertension, decision to treat and intensification of treatment and therefore SBP is often forgotten and-or misinterpreted in this decision making process. On the other hand, since current antihypertensive drugs are equally effective in lowering SBP and DBP and-or less effective in lowering SBP more than DBP, SBP is often uncontrolled in treated patients with isolated systolic hypertension or prevalent increase in SBP. The possibility of obtaining better control of SBP in the future is linked to better education of physicians, who need to pay greater attention to SBP as a parameter for diagnosing, treating and intensifying treatment, and to the development of new drugs more active in reducing SBP. PMID- 14529554 TI - Mechanisms of endothelial dysfunction: clinical significance and preventive non pharmacological therapeutic strategies. AB - Endothelium-derived NO is not only a potent vasodilator but also inhibits platelet aggregation, vascular smooth muscle cell migration and proliferation, monocyte adhesion and adhesion molecule expression, thus protecting the vessel wall against the development of atherosclerosis. Cardiovascular risk factors are associated with an imbalance of the redox equilibrium towards oxidative stress and, therefore, impair the integrity of the endothelium, leading to endothelial activation which involves blunted endothelium-dependent vasodilation (vasodilator dysfunction) as well as inflammatory processes extending to the milieu within the whole vasculature, making plaques prone to rupture. In prospective studies endothelial dysfunction is associated with increased incidence of cardiovascular events. Thus, the prevention of endothelial dysfunction can determine a strong advantage in the clinical outcome of patients with cardiovascular risk factors. Several non-pharmacological interventions can prevent endothelial dysfunction or improve impaired endothelium-dependent vasodilation. Probably the most effective non-pharmacological measure is represented by aerobic physical activity, which can reduce production of oxidative stress associated to increasing age. Moreover, physical activity can improve endothelial dysfunction even in patients with cardiovascular risk factors such as essential hypertension. In addition several other approaches, including vitamin and fish oil supplementation, or tea and red wine consumption, can lead to an improvement of endothelium-dependent vasodilation, possibly by a restoration of NO availability. It is worth noting that most of non-pharmacological measures act by preventing or reducing oxidative stress. PMID- 14529555 TI - New technological developments in the clinical imaging of atherosclerotic plaque. AB - Direct visualization of the composition of the atherosclerotic plaque during its natural history and after therapeutic intervention may be helpful in detecting lesions with high risk of acute events and in understanding progression and regression of the disease. A wide variety of invasive and non-invasive imaging techniques is available to detect clue aspects of atherosclerosis from the early stage to the clinical evidence appearance. We will firstly review the ongoing technological and clinical research on both invasive and non-invasive techniques. Afterward, we will discuss in detail the use of high-resolution, multi-contrast magnetic resonance imaging for non-invasive imaging of the plaque and its characterization in terms of its various components (i.e., thickness, lipid, fibrous, calcium, or thrombus). Finally, we will describe the potential of quantitative analysis in describing of plaque constituents with improved reproducibility. PMID- 14529556 TI - Relationship between cigarette smoking and other coronary risk factors in atherosclerosis: risk of cardiovascular disease and preventive measures. AB - Among the major Coronary Risk Factors (CRF) cigarette smoking has shown undoubtedly harmful effects on the heart and blood vessels either as active smoking (smoking a cigarette) or passive smoking (exposure to environmental tobacco smoke -ETS). The strong relationship between cigarette smoking and cardiovascular disease has been seen independent of the other CRF in a number of well-designated epidemiologic studies. However, a strong increase in the excess of cardiovascular risk has been defined along with the interaction of cigarette smoking and other major CRF. Thousands of pharmacologically active substances are present in tobacco smoke, and a large number of direct and indirect effects have been demonstrated. Different responses are also related to these types of exposure: active exposure or passive exposure. The cardiovascular risk increases with increasing levels of blood pressure and/or serum cholesterol and diabetes mellitus, and at each level of these three risk factors, distributed with different rates according to age and gender in individuals, the risk in active smokers or passive smokers is greater than the risk in nonsmokers. Further analytical and methodological observations are needed for better understanding of the chemical and biological synergism. Nevertheless, evidence is clear that cigarette smoking greatly increases the risk of cardiovascular diseases in individuals already at increased risk because of other CRF. Preventive measures must be absolutely conducted to prevent the CRF interaction. These are the changes in lifestyle (i.e. to give up smoking and make physical activity), drug administration, diet supplementation especially by those substances with antioxidant effects. PMID- 14529557 TI - The effects of lipid-regulating therapy on haemostatic parameters. AB - There is growing evidence that the components of the haemostatic system play a significant role in the development and progression of atherosclerosis and its complications. Lipid-lowering interventions have been associated with a significant reduction of morbidity and mortality. However, the improvement in cardiovascular risk seen in several clinical trials is incompletely explained by cholesterol reduction. Therefore, the benefit from lipid lowering drugs may involve non-lipid mechanisms. These include beneficial effects on the arterial wall, improved endothelial function and a favourable influence on blood rheology and thrombogenesis. In this review, we consider the influence of lipid-lowering interventions on rheological and haemostatic parameters as well as the potential clinical relevance of these effects. PMID- 14529558 TI - The effects of antihypertensive therapy on haemostatic parameters. AB - There is extensive trial-based evidence showing that antihypertensive drugs reduce the risk of vascular events (e.g. stroke and myocardial infarction) as well as target organ damage (e.g. left ventricular hypertrophy and microalbuminuria). However, some of these benefits appear to be, at least partially, independent of the extent of blood pressure (BP) lowering. It is also evident that in certain clinical situations some antihypertensive drugs are more effective than others. In this review we discuss the effects of antihypertensive drugs on the endothelium, platelets, fibrinolysis and coagulation. These properties may account for the observed BP-independent actions. Antihypertensive drugs exert multiple effects on the vascular endothelium. These include effects on nitric oxide (NO) and angiotensin II-mediated actions. Many BP lowering drugs can inhibit platelet activity, although the relevance of this property is unknown, especially if patients are also taking platelet inhibitors (e.g. aspirin). Antihypertensive drugs also influence fibrinolysis and coagulation. These effects may be mediated by a variety of mechanisms, including altering insulin sensitivity. The haemostatic actions of antihypertensive drugs deserve greater recognition and further investigation. PMID- 14529559 TI - Coronary risk factors in the elderly: their interactions and treatment. AB - Coronary disease is currently a major cause of disability and mortality in older populations. Due to several factors--including increased noncardiovascular mortality, atypical presentation of coronary events, selective survival, and, possibly, clustering of protective genetic traits--the relative risk associated with almost all risk factors decreases with advancing age. Nonetheless, all available evidences from both observational studies and randomized trials indicate that, due to the higher event rates, the absolute risk reduction yielded by preventive interventions is much greater in the older age segments of populations. Another implication of the complex relationships between risk factors and comorbid conditions in the pathogenesis of coronary-related events and mortality, typical of the elderly subjects, is represented by the multiple effects of treatment for single risk factors, such as the decrease in LDL cholesterol levels and inflammation markers yielded by statins. Taken together, these factors account for the more favorable cost-effectiveness ratios of preventive interventions in the older, as compared with the middle-aged subjects. On the other hand, the high level of interaction between coexistent risk factors and comorbidity renders a global approach to the prevention of coronary events in older subjects mandatory for physicians, as well as for decision-makers. In fact, a multidimensional assessment--including the evaluation of cognitive, affective and social disturbances--driving a multidisciplinary treatment of risk factors- encompassing behavioral counseling and social support--is essential to improve patients' compliance and to effectively reduce the burden of coronary-related morbidity and mortality in older populations. PMID- 14529560 TI - Clock genes in calendar cells as the basis of annual timekeeping in mammals--a unifying hypothesis. AB - Melatonin-based photoperiod time-measurement and circannual rhythm generation are long-term time-keeping systems used to regulate seasonal cycles in physiology and behaviour in a wide range of mammals including man. We summarise recent evidence that temporal, melatonin-controlled expression of clock genes in specific calendar cells may provide a molecular mechanism for long-term timing. The agranular secretory cells of the pars tuberalis (PT) of the pituitary gland provide a model cell-type because they express a high density of melatonin (mt1) receptors and are implicated in photoperiod/circannual regulation of prolactin secretion and the associated seasonal biological responses. Studies of seasonal breeding hamsters and sheep indicate that circadian clock gene expression in the PT is modulated by photoperiod via the melatonin signal. In the Syrian and Siberian hamster PT, the high amplitude Per1 rhythm associated with dawn is suppressed under short photoperiods, an effect that is mimicked by melatonin treatment. More extensive studies in sheep show that many clock genes (e.g. Bmal1, Clock, Per1, Per2, Cry1 and Cry2) are expressed in the PT, and their expression oscillates through the 24-h light/darkness cycle in a temporal sequence distinct from that in the hypothalamic suprachiasmatic nucleus (central circadian pacemaker). Activation of Per1 occurs in the early light phase (dawn), while activation of Cry1 occurs in the dark phase (dusk), thus photoperiod induced changes in the relative phase of Per and Cry gene expression acting through PER/CRY protein/protein interaction provide a potential mechanism for decoding the melatonin signal and generating a long-term photoperiodic response. The current challenge is to identify other calendar cells in the central nervous system regulating long-term cycles in reproduction, body weight and other seasonal characteristics and to establish whether clock genes provide a conserved molecular mechanism for long-term timekeeping. PMID- 14529561 TI - Macrophage migration inhibitory factor: a neuroendocrine modulator of chronic inflammation. AB - The diverse actions of macrophage migration inhibitory factor (MIF) within the immuno-neuroendocrine system are yet to be fully understood, but it is clear that MIF plays a pivotal role in the regulation of both the innate and adaptive immune response. An emerging body of data presently indicates that MIF's position within the cytokine cascade is to act in concert with glucocorticoids to control the 'set point' and magnitude of the immune and inflammatory response. In this article we will review the actions of MIF within the immune system and discuss the overlapping and contrasting aspects of MIF and glucocorticoid biology. In particular we will focus on the role of MIF within the immuno-neuroendocrine interface and suggest molecular mechanisms by which MIF may counter-regulate glucocorticoid function. Finally we will discuss emerging evidence that functional MIF gene-promoter polymorphisms render one susceptible to elevated MIF expression, and the development of an exaggerated immune/inflammatory response that potentiates the progression to chronic inflammatory disease. PMID- 14529562 TI - Phosphatidylinositol 3-kinase in cumulus cells is responsible for both suppression of spontaneous maturation and induction of gonadotropin-stimulated maturation of porcine oocytes. AB - In this study, we investigated the mechanisms of protein kinase B (PKB) activation and its role in cumulus cells during in vitro meiotic resumption of porcine oocytes. PKB activity in cumulus cells was significantly decreased by 12 h cultivation of cumulus-oocyte complexes (COCs) in basic medium. However, the addition of phosphodiesterase inhibitors, hypoxanthine or 3-isobutyl-1 methylxanthine, maintained the level of PKB activity in cumulus cells at comparable with that in cumulus cells just after collection from their follicles. When COCs were cultured with phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor, LY294002, PKB activity was significantly decreased, and both caspase 3 activity and the proportion of apoptotic cells were significantly increased as compared with those in cumulus cells just after collection from their follicles. Moreover, the inhibitory effect of hypoxanthine on spontaneous meiotic resumption was overcome by addition of LY294002. On the other hand, markedly high activity of PKB and high intensity of the phosphorylated PKB band were observed in cumulus cells of COCs which were cultured with FSH. The addition of 20 microM LY294002 to FSH-containing medium induced an apoptosis of cumulus cells, whereas little apoptotic-positive signal was detected in COCs cultured with 5 microM LY294002 and FSH. However, the inhibitory effects of LY294002 on progesterone production by cumulus cells and germinal vesicle breakdown in oocytes reached a maximum at 5 microM. Thus, high activity of the PI 3-kinase-PKB pathway in cumulus cells plays an important role in FSH regulation of cell function. Judging from these results, it is estimated that PI 3-kinase in cumulus cells is required for both the suppression of spontaneous meiotic resumption and the induction of gonadotropin stimulated meiotic resumption. PMID- 14529563 TI - Gonadotropin treatment increases homocysteine levels in idiopathic hypogonadotropic hypogonadism: an indirect effect mediated by changes in body composition. AB - The main objective of the present study was to examine the alterations in plasma total homocysteine (tHcy) concentrations during a testosterone-deficient state and after gonadotropin treatment for 6 Months in patients with idiopathic hypogonadotropic hypogonadism (IHH). Thirty-five newly diagnosed male patients with IHH (mean age 21.34+/-1.53 years) and 29 age- and body mass index-matched healthy males (mean age 21.52+/-1.77 years) were recruited into the study. Pretreatment levels of free testosterone (1.51+/-0.66 pg/ml), estradiol (21.37+/- 4.37 pg/ml), FSH (0.91+/-0.24 IU/l) and LH (1.25+/- 0.53 IU/l) were lower than controls (25.17+/-3.06 pg/ml, 31.00+/-4.96 pg/ml, 3.14+/-1.62 IU/l and 4.83+/ 1.65 IU/l respectively) (P<0.001). They increased significantly after treatment (18.18+/-1.59 pg/ml, 27.97+/- 4.25 pg/ml, 2.41+/-0.27 IU/l and 2.79+/-0.19 IU/l respectively) (P<0.001). Patients with IHH had lower tHcy levels than controls (10.14+/-1.34 and 12.58+/- 2.29 micro mol/l respectively) (P<0.001). Plasma tHcy concentrations increased significantly (12.63+/-1.44 micromol/l) after 6 months of treatment (P<0.001). As compared with the controls, pretreatment levels of serum creatinine (63.54+/-13.01 vs 82.84+/-16.69 micromol/l), hemoglobin (12.98+/ 0.56 vs 13.83+/-0.71 g/dl) and hematocrit (39.29+/-2.01 vs 41.38+/-1.95%) were significantly lower (P<0.001), and they increased significantly following treatment (80.24+/-11.93 micromol/l, 13.75+/-0.49 g/dl and 41.26+/-1.78% respectively) (P<0.001). The pretreatment folic acid and vitamin B(12) levels were significantly higher in patients when compared with controls (14.87+/-5.68 vs 12.52+/-4.98 nmol/l, P=0.034 and 289.75+/-92.34 vs 237.59+/-108.17 pmol/l, P=0.002 respectively). They decreased significantly after treatment (11.29+/-3.31 nmol/l and 228.51+/-54.33 pmol/l respectively) (P<0.001). The univariate and multivariate regression analysis results showed that only changes in creatinine, creatinine clearance, vitamin B12 and folic acid were independently associated with changes in tHcy levels in patients with IHH. In conclusion, the increase in plasma tHcy concentrations following gonadotropin treatment seems to be largely independent of changes in androgen levels. PMID- 14529564 TI - Oestrogen receptor alpha increases p21(WAF1/CIP1) gene expression and the antiproliferative activity of histone deacetylase inhibitors in human breast cancer cells. AB - We analysed the antiproliferative activity of various histone deacetylase (HDAC) inhibitors such as trichostatin A (TSA) on human breast cancer cells. We observed a lower sensitivity to HDAC inhibition for oestrogen receptor negative (ER-) versus positive (ER+) cell lines. This differential response was associated neither with a modification of drug efflux via the multidrug resistance system nor with a global modification of histone acetyltransferase (HAT)/HDAC activities. In contrast, we demonstrated that in ER+ breast cancer cells the p21(WAF1/CIP1) gene was more sensitive to TSA regulation and was expressed at higher levels. These differences were observed both in transient transfection experiments and on the endogenous p21(WAF1/CIP1) gene. The Sp1 transcription factor, which was shown to interact in vitro with both class I and class II HDACs, is sufficient to confer the differential sensitivity to TSA and participated in the control of p21(WAF1/CIP1) basal expression. Finally, re expression of ERalpha following adenoviral infection of ER- breast cancer cells increased both p21(WAF1/CIP1) protein accumulation and the growth inhibitory activity of TSA. Altogether, our results highlight the key role of ERalpha and p21(WAF1/CIP1) gene expression in the sensitivity of breast cancer cells to hyperacetylating agents. PMID- 14529565 TI - Overexpression of wild-type p53 gene renders MCF-7 breast cancer cells more sensitive to the antiproliferative effect of progesterone. AB - We have recently shown that growth inhibition of breast cancer cells by progesterone is due to the induction of cell differentiation, but not apoptosis. Because the tumor suppressor protein p53 plays a central role in normal cell growth and in tumor suppression, we have examined the effect of progesterone on the levels of this protein in MCF-7 cells. We show here that the antiproliferative effect of progesterone is accompanied with down-regulation of endogenous p53 protein. To study the effect of progesterone on cell growth in the presence of normal levels of p53 protein, we used transient transfection to overexpress p53 protein. MCF-7 cells were transfected with a p53 expressing vector that contains p53 human cDNA under the control of a cytomegalovirus promoter. Cell growth, cell viability, and apoptosis were analyzed in the transfected cells after six days of exposure to 100 nM progesterone. We show here that progesterone significantly enhances growth inhibition and apoptosis in MCF-7 cells overexpressing p53, but not in cells transfected with the control vector. These data suggest that re-establishing p53 function in MCF-7 breast cancer cells renders them more sensitive to the growth inhibitory effect of progesterone. PMID- 14529566 TI - Expression and regulation of glucose transporter 8 in rat Leydig cells. AB - Basal and LH/human chorionic gonadotropin (hCG)-stimulated testosterone formation by Leydig cells is dependent on ambient glucose levels. Inhibition of glucose uptake is associated with decreased testosterone formation. Recently, glucose transporter 8 (GLUT8) has been shown to be highly expressed in the testis. In the present study, we have investigated the expression and regulation of the GLUT8 gene in rat Leydig cells. Primers were designed by using sequences that are not conserved in GLUT1 to GLUT5 and that contain the glycosylation region of GLUT8. This yielded an amplicon of 186 bp. The tIssue-specific expression experiments in adult rat (55- to 65-day-old) tIssues revealed that GLUT8 is expressed predominantly in the testis, in smaller amounts in heart and kidney, and in negligible amounts in liver and spleen. Furthermore, GLUT8 mRNA was found to be highly expressed in crude interstitial cells, Leydig cells and testicular and epididymal germ cells. In prepubertal rat (20-day-old) tIssues, GLUT8 expression was comparatively much lower than in the adult rat tIssues. By comparative RT PCR, hCG caused dose- and time-dependent increases of GLUT8 mRNA levels. hCG and IGF-I had synergistic effects on GLUT8 mRNA and protein expression. GLUT1 and GLUT3 were also found to be expressed in Leydig cells. However, neither GLUT1 nor GLUT3 were affected by treatments with hCG, IGF-I or hCG and IGF-I combined. The addition of murine interleukin-1alpha (mIL-1alpha; 10 ng/ml), murine tumor necrosis factor-alpha (mTNF-alpha; 10 ng/ml), murine interferon-gamma (mIFN gamma; 500 U/ml) separately or in combination decreased hCG-induced GLUT8 mRNA levels significantly. In conclusion, GLUT8 mRNA in Leydig cells was positively regulated by hCG and IGF-I and down-regulated by cytokines, mIL-1alpha, mTNF alpha and mIFN-gamma. These results indicate that hCG, growth factors and cytokines affect Leydig cell steroidogenesis by modulating GLUT8 expression. PMID- 14529567 TI - Frequent overexpression of cyclin D1 in sporadic pancreatic endocrine tumours. AB - Pancreatic endocrine tumours (PETs) occur sporadically or are inherited as part of the multiple endocrine neoplasia type-1 syndrome. Little is known about the molecular events leading to these tumours. Cyclin D1, a key regulator of the G1/S transition of the cell cycle, is overexpressed in a variety of human cancers as well as certain endocrine tumours. We hypothesized that similar to other endocrine tumours, cyclin D1 is overexpressed in human sporadic PETs. Cyclin D1 protein overexpression was found in 20 of 31 PETs (65%) when compared with normal pancreatic tIssue. Furthermore, Northern blot analysis suggests that cyclin D1 up regulation occurs at the post-transcriptional level in some PETs. Because the key cell growth signalling pathways p42/p44/ERK (extracellular signal-regulated kinase), p38/MAPK (mitogen-activated protein kinase), and Akt/PKB (protein kinase B) can regulate cyclin D1 protein expression in other cell types, pancreatic endocrine tumours were analysed with phospho-specific antibodies against the active forms of these proteins to elucidate a tIssue-specific regulatory mechanism of cyclin D1 in PETs. We found frequent activation of the p38/MAPK and Akt pathways, but down-regulation of the ERK pathway, in cyclin D1 overexpressing PETs. This study demonstrates that cyclin D1 overexpression is associated with human sporadic PET tumorigenesis, and suggests that this up-regulation may occur at the post-transcriptional level. These findings will direct future studies of PETs towards cell cycle dysregulation and the identification of key growth factor pathways involved in the formation of these tumours. PMID- 14529568 TI - Angiotensinogen localization and secretion in the rat pancreas. AB - Renin and angiotensinogen have been previously found in the rat pancreas, and angiotensin receptors have been located in the apical domain of duct cells. To evaluate the possibility that angiotensin II could be generated within the duct system, we decided to determine whether angiotensinogen is present in rat pancreatic juice and the angiotensinogen-immunoreactive pancreatic cell types that could be responsible for its production. Angiotensinogen was detected in significant amounts by Western blotting in pancreatic juice collected from several individual rats. Different isoforms between plasma and pancreatic juice angiotensinogens were demonstrated by isoelectric focusing. Immunocytochemical experiments revealed angiotensinogen-immunoreactive cells at the periphery of the islets of Langerhans, and confocal microscopy demonstrated that most angiotensinogen-immunoreactive cells were glucagon-secreting cells. Secretion of angiotensinogen did not follow the regulated secretory pathway since it was absent from the glucagon-containing granules. This was confirmed by electron microscopy immunocytochemistry. Duct and acinar cells did not express angiotensinogen at an immunocytochemical detectable level. The present findings indicated an exocrine secretion of angiotensinogen by glucagon-secreting cells and suggest that one of the final targets of the local pancreatic renin angiotensin system may be the duct epithelium. PMID- 14529569 TI - Liver 5'-deiodinase activity is modified in rats under restricted feeding schedules: evidence for post-translational regulation. AB - Restricted feeding schedules (RFSs) produce a behavioral activation known as anticipatory activity, which is a manifestation of a food-entrained oscillator (FEO). The liver could be playing a role in the physiology of FEO. Here we demonstrate that the activity of liver selenoenzyme deiodinase type 1 (D1), which transforms thyroxine into triiodothyronine (T3), decreases before food access and increases after food presentation in RFSs. These changes in D1 activity were not due to variations in D1 mRNA. In contrast, a 24 h fast promoted a decrease in both D1 activity and mRNA content. The adjustment in hepatic D1 activity was accompanied by a similar modification in T3-dependent malic enzyme, suggesting that the local generation of T3 has physiological implications in the liver. These results support the notion that the physiological state of rats under RFSs is unique and distinct from rats fed freely or fasted for 24 h. Data also suggest a possible role of hepatic D1 enzyme in coordinating the homeorhetic state of the liver when this organ participates in FEO expression. PMID- 14529570 TI - Ghrelin-induced GH secretion in domestic fowl in vivo and in vitro. AB - Although avian and mammalian species differ significantly in their regulation of GH secretion, preliminary studies have demonstrated in vivo GH responses to ghrelin in chickens, as in mammals. However, the relative potency of ghrelin as a GH-releasing hormone (GHRH) in birds is uncertain, as is its site of action. The intravenous administration of human ghrelin to immature chickens promptly increased the circulating GH concentration (within 10 min), although this was transitory and was only maintained for 20 min. This GH response was dose-related with an EC50 of approximately 3.0 microg/kg, comparable with the reported potency of human GHRH in chickens. When incubated with dispersed pituitary cells, human ghrelin induced dose-dependent GH release over a range of 10(-6) to 10(-9) M, with an EC50 of 7.0 x 10(-8) M, comparable with that induced by human GHRH (EC50 6.0 x 10(-8) M), although it was less effective at doses of 10(-6) to 10(-8) M. This was due to direct effects on pituitary somatotrophs, since human ghrelin increased GH release (determined by the reverse hemolytic plaque assay) from individual pituitary cells. The incubation of these cells with human ghrelin induced a dose-dependent increase in the numbers of somatotrophs secreting GH and in the amount of GH released by each cell. In summary, these results demonstrated that ghrelin is a dose-related GH-releasing factor in chickens with a potency comparable with that induced by human GHRH. The GH-releasing action of ghrelin is due, at least in part, to stimulatory actions on the numbers of somatotrophs induced to release GH and upon the amount of GH released from individual somatotrophs. PMID- 14529571 TI - Endotoxin at low doses stimulates pituitary GH whereas it decreases IGF-I and IGF binding protein-3 in rats. AB - While it is well known that sepsis inhibits serum IGF-I and its gene expression in the liver, the effect on pituitary GH and IGF-binding protein-3 (IGFBP-3) is poorly understood. The GH-IGF-I-IGFBP-3 response to different doses of lipopolysaccharide (LPS) administration has been investigated in adult male rats. Two experiments were performed, administration of low doses of LPS (5, 10, 50 and 100 microg/kg) and high doses of LPS (100, 250, 500 and 1000 microg/kg). Rats received two i.p. injections of LPS (at 1730 h and 0830 h the following day) and were killed 4 h after the second injection. LPS administration induced a biphasic response in serum concentrations of GH, with an increase at the 10 microg/kg dose, followed by a decrease at higher doses (100 microg/kg on up). Pituitary GH mRNA was also increased by the administration of 10 and 50 microg/kg LPS, whereas at higher doses LPS did not modify pituitary GH mRNA. We also analyzed the GH response to LPS in primary pituitary cell cultures. When exposed to LPS, in the culture medium, there was an increase in GH release at the concentration of 0.1 and 10 ng/ml, whereas more concentrated LPS did not modify GH release. Serum concentrations of IGF-I declined in a dose-dependent fashion after LPS administration in the rats injected with 10 microg/kg LPS on up. This decrease is secondary to modifications in its synthesis in the liver, since endotoxin injection decreased both IGF-I and its mRNA in the liver. The liver GH receptor mRNA was also decreased by LPS administration, but only in the animals injected with high LPS doses. There was a decrease in both the IGFBP-3 serum levels and its gene expression in the liver with all LPS doses studied. These data suggest a biphasic LPS effect on pituitary GH, a stimulatory effect at low doses and an inhibitory effect at higher doses, whereas it has a clear inhibitory effect on IGF-I and IGFBP-3 synthesis in the liver. The decrease in liver IGFBP-3 mRNA and in serum concentrations of IGFBP-3 in the rats injected with LPS may contribute to the decrease in serum concentrations of IGF-I. PMID- 14529572 TI - Distribution analysis of vitamin D highlights differences in population subgroups: preliminary observations from a pilot study in UK adults. AB - There is no consensus between Authors on the definition of a replete or deficient vitamin D state. Our aim was to describe a suitable method that could be used to compare vitamin D data in subject groups with small or large numbers. Two hundred and forty indigenous asymptomatic, non-pregnant adult subjects recruited from a single-consultation outpatient attendance with normal biochemistry, represented a sample of our inner city district population. 25-hydroxyvitamin D (25,OHD3) levels were measured to illustrate the effects of season, sex and ethnic group on vitamin D levels and subjected to distribution analysis. This method quantifies as a percentage the distribution of 25,OHD3 concentrations (observed concentration, OC) in pooled group data. The data can be expressed as distribution frequency domains or cumulative frequency ogives (0-100%) or transformed into discrete linear probits, amenable to regression analysis. An estimate of the OC50 (mid-point) and upper (either OC75 or OC95) or lower (either OC25 or OC5) range or at any other frequency between subject groups can be compared. A marked difference in 25,OHD3 levels between Asian and non-Asian asymptomatic adult subjects was seen during both seasons. 25,OHD3 deficiency was defined as at or below the OC25 for the non-Asian group (for both sexes: winter < 13.36 ng/ml, summer <13.38 ng/ml). The majority of Asians of both sexes were 25,OHD3 deficient (winter 94%, summer 82%). The distribution analysis provides an easy technique to compare 25,OHD3 status of different subject groups, allowing the description of populations using either longitudinal or cross-sectional data. This method may offer a way of describing 25,OHD3 deficiency between observers worldwide. PMID- 14529573 TI - Dexamethasone does not exert direct intracellular feedback on steroidogenesis in human adrenal NCI-H295A cells. AB - Experimental therapy of fetuses affected with congenital adrenal hyperplasia (CAH) has been reported by administering dexamethasone (Dex) to pregnant women at risk for carrying a CAH fetus. Such prenatal therapy can almost wholly eliminate virilization of the external genitalia of affected female fetuses, but only when treatment is started before 9 weeks of gestation. As it is not known whether the hypothalamic-pituitary-adrenal axis is functional at this time, and as the minimal effective doses of Dex are substantially supraphysiologic for the fetus, the mechanism of action of prenatal Dex treatment has been unclear. To assess the possibility that Dex might act directly on the adrenal, we used human adrenocortical NCI-H295A cells, an established model of the human fetal adrenal. Short term (6 h) incubation of these cells with Dex decreased synthesis of 11 deoxycortisol and 17alpha-hydroxyprogesterone and increased synthesis of deoxycorticosterone (DOC), but only at very high concentrations of Dex (> or =10 microM) that affect P450c17 (17alpha-hydroxylase/17,20 lyase) as a competitive inhibitor; no effects were seen at lower concentrations corresponding to those used in prenatal treatment. When NCI-H295A cells were treated with up to 100 microM Dex for 72 h, dot-blot analysis showed no changes in the abundance of the mRNAs for P450scc (cholesterol side-chain cleavage enzyme), P450c17, or 3beta hydroxysteroid dehydrogenase II (3betaHSDII). When NCI-H295A cells were transfected with promoter/reporter constructs for the human genes for P450scc and P450c17, 24-h treatment with Dex at doses up to 100 microM had no effect on the transcription of these constructs. Because the regulation of steroidogenic processes in NCI-H295A cells closely resembles such regulation in the adrenal, we suggest that Dex may not act by an intra-adrenal mechanism in the experimental prenatal treatment of CAH. PMID- 14529574 TI - Effect of topical fluoroquinolones on the expression of matrix metalloproteinases in the cornea. AB - BACKGROUND: Matrix metalloproteinases play an important role in extracellular matrix deposition and degradation. Based on previous clinical observations of corneal perforations during topical fluoroquinolone treatment, we decided to evaluate the comparative effects of various fluoroquinolone eye drops on the expression of matrix metalloproteinases (MMPs) in cornea. METHODS: Eighty female Lewis rats were divided into two experimental groups: intact and wounded corneal epithelium. Uniform corneal epithelial defects were created in the right eye with application of 75% alcohol in the center of the tissue for 6 seconds. The treatment groups were tested as follows: 1) Tear drops: carboxymethylcellulose sodium 0.5 % (Refresh, Allergan); 2) Ciprofloxacin 0.3% (Ciloxan, Alcon); 3) Ofloxacin 0.3%(Ocuflox, Allergan); 4) Levofloxacin 0.5%(Quixin, Santen). Eye drops were administered 6 times a day for 48 hours. Rats were sacrificed at 48 hours. Immunohistochemical analysis and zymography were conducted using antibodies specific to MMPs-1, 2, 8 and 9. RESULTS: MMP-1, MMP-2, MMP-8 and MMP-9 expression were detected at 48 hrs in undebrided corneal epithelium groups treated with the topical fluoroquinolones. No statistical difference was observed in quantitative expression of MMPs among ciprofloxacin 0.3%, ofloxacin 0.3%, levofloxacin 0.5%. When the artificial tear group and the fluoroquinolone groups with corneal epithelial defect were compared, increased expression of MMPs was observed as a result of the wound healing process. However, the fluoroquinolone treated group exhibited high statistically significantly levels of MMPs expression. CONCLUSIONS: Our study provides preliminary evidence that topical application of fluoroquinolone drugs can induce the expression of MMP-1, MMP-2, MMP-8 and MMP-9 in the undebrided corneal epithelium compared to artificial tear eye drops. PMID- 14529576 TI - Flow-mediated release of nitric oxide from lymphatic endothelial cells of pressurized canine thoracic duct. AB - We examined the effects of flow on lymphatic endothelial cells by using conventional cascade preparations of isolated coronary arteries without intact endothelium. The pressurized thoracic ducts were intraluminally perfused at a constant flow rate ranging from 0.5 to 2.0 ml/min. A linear relationship was observed between the flow rate and the normalized amount of relaxing substance(s) released from the lymphatic endothelial cells. Thus the flow rate of 2.0 ml/min produced approximately 39% of sodium nitroprusside (SNP)-produced maximal relaxation in the cascade arterial rings. The acetylcholine (ACh, 10(-5) M)- and flow-induced relaxations of the cascade arterial rings were completely reduced by the mechanical rubbing of lymphatic endothelial cells in the pressurized lymph vessels. Pretreatment with 5 x 10(-5) M N(G)-nitro-L-arginine methyl ester (L NAME) on the lymphatic endothelial cells caused a significant reduction of the ACh- and flow-induced vasodilations of the cascade arterial rings. Pretreatment with 10(-5) M indomethacin on the lymphatic endothelial cells produced no significant effect on the ACh- and flow-induced vasodilations. These findings suggest that lymphatic endothelial cells of canine thoracic ducts can produce and release endogenous nitric oxide by stimulation of flow (approximately 2.0 ml/min). PMID- 14529577 TI - The gating and conductance properties of Cav3.2 low-voltage-activated T-type calcium channels. AB - Calcium channels are essential for excitation-contraction coupling and pacemaker potentials in cardiac muscle cells. Whereas L-type Ca(2+) channels have been extensively studied, T-type channels have been poorly characterized in cardiac myocytes. We describe here the functional properties of recombinant Ca(V)3.2 T type Ca(2+) channels expressed in mammalian cell lines. The T-type Ca(2+) current showed a rapid activation and an inactivation phase in response to depolarization, and it displayed a window current over the voltage range from -60 to -40 mV in 1 to 10 mM external Ca(2+). Barium (Ba(2+)) and strontium (Sr(2+)) permeate the channel with similar activation kinetics. On the other hand, monovalent cations, Li(+) and Na(+), permeate the T-type Ca(2+) channel more easily than the L-type Ca(2+) channel. The permeability order of the Ca(V)3.2 T type Ca(2+) channel among monovalent and divalent cations was determined as Ba(2+)>Mn(2+)>Ca(2+)>Sr(2+)>Li(+1)>Na(+) with the permeability order of 1.39:1.25:1.00:0.95:0.55:0.29. The ionic conductance sequence for cations relative to calcium was Sr(2+)>Ba(2+)>Ca(2+)>Li(+1)>Mn(2+)>Na(+) with the conductance ratio of 1.39:1.21:1.00:0.40:0.23:0.11. The permeation profile of manganese (Mn(2+)) is complex. Mn(2+) permeates the Ca(2+) channel with a permeability similar to Ca(2+) or Ba(2+), but with a much smaller current density, resulting in a much smaller conductance. The properties relating to progression and recovery from inactivation in the Ca(V)3.2 channel are substantially identical with either Ca(2+) or Ba(2+) as the charge carrier. PMID- 14529578 TI - Differential activation of protein kinase C between ischemic and pharmacological preconditioning in the rabbit heart. AB - The objective of the present study was to investigate the differential activation of protein kinase C between ischemic (IPC) and pharmacological preconditioning (PPC) in the rabbit heart. Control, IPC, diazoxide (Diaz), and chelerythrine (Chel)+IPC groups underwent prolonged coronary artery occlusion (CAO) for 30 minutes followed by 180 minutes' reperfusion (protocol I). In protocol II, sham, IPC-only, Diaz-only, and Chel+IPC-only groups did not undergo prolonged CAO. IPC was induced with 4 cycles of 5-min regional ischemia and 10-min reperfusion before prolonged CAO. Diaz (5 mg/kg) was administered 30 min before prolonged CAO. Chel (5 mg/kg) was administered 5 min before the IPC procedure. Infarct size was determined by tetrazolium staining. Assessment of protein kinase C (PKC) isoforms from a left ventricular (LV) sample was conducted by western blotting. Apoptosis in situ was determined by TUNEL assay. The infarction area in the IPC (11.6 +/- 1.0%) and Diaz (19.5 +/- 3.8%) groups was reduced significantly (p< 0.01, p< 0.05) relative to the control group (40.0 +/- 3.8%). The reduction by IPC was abolished by pretreatment with Chel. Apoptosis was significantly decreased (p< 0.01) in the IPC and diazoxide groups compared with the control and Chel+IPC groups (control: 4.78 +/- 0.56% vs. IPC: 2.00 +/- 0.38% vs. Diaz: 2.20 +/- 0.32% vs. Chel+IPC: 4.32 +/- 0.41%) and DNA laddering was attenuated in the IPC and Diaz groups. Membrane PKC-epsilon levels in the IPC and Diaz groups increased significantly relative to the control and Chel+IPC groups. Membrane PKC epsilon levels in the IPC-only group showed greater increases than the Diaz-only and Chel+IPC-only groups. These findings suggest that whereas PPC suppresses apoptosis when diazoxide opens mitochondrial K(ATP) channels and then activates PKC-epsilon through ischemia-reperfusion, IPC activates PKC-epsilon in the particulate fraction prior to continuous ischemia-reperfusion. We concluded that the difference between IPC and PPC appears to consist in the difference in the timing of PKC-epsilon activation, though both IPC and PPC provide the cardioprotection in ischemia-reperfusion injury. PMID- 14529575 TI - Follow-up of patients with curatively resected colorectal cancer: a practice guideline. AB - BACKGROUND: A systematic review was conducted to evaluate the literature regarding the impact of follow-up on colorectal cancer patient survival and, in a second phase, recommendations were developed. METHODS: The MEDLINE, CANCERLIT, and Cochrane Library databases, and abstracts published in the 1997 to 2002 proceedings of the annual meeting of the American Society of Clinical Oncology were systematically searched for evidence. Study selection was limited to randomized trials and meta-analyses that examined different programs of follow-up after curative resection of colorectal cancer where five-year overall survival was reported. External review by Ontario practitioners was obtained through a mailed survey. Final approval of the practice guideline report was obtained from the Practice Guidelines Coordinating Committee. RESULTS: Six randomized trials and two published meta-analyses of follow-up were obtained. Of six randomized trials comparing one follow-up program to a more intense program, only two individual trials detected a statistically significant survival benefit favouring the more intense follow-up program. Pooling of all six randomized trials demonstrated a significant improvement in survival favouring more intense follow up (Relative Risk Ratio 0.80 (95%CI, 0.70 to 0.91; p = 0.0008). Although the rate of recurrence was similar in both of the follow-up groups compared, asymptomatic recurrences and re-operations for cure of recurrences were more common in patients with more intensive follow-up. Trials including CEA monitoring and liver imaging also had significant results, whereas trials not including these tests did not. CONCLUSION: Follow-up programs for patients with curatively resected colorectal cancer do improve survival. These follow-up programs include frequent visits and performance of blood CEA, chest x-rays, liver imaging and colonoscopy, however, it is not clear which tests or frequency of visits is optimal. There is a suggestion that improved survival is due to diagnosis of recurrence at an earlier, asymptomatic stage which allows for more curative resection of recurrence. Based on this evidence and consideration of the biology of colorectal cancer and present practices, a guideline was developed. Patients should be made aware of the risk of disease recurrence or second bowel cancer, the potential benefits of follow-up and the uncertainties requiring further clinical trials. For patients at high-risk of recurrence (stages IIb and III) clinical assessment is recommended when symptoms occur or at least every 6 months the first 3 years and yearly for at least 5 years. At the time of those visits, patients may have blood CEA, chest x-ray and liver imaging. For patients at lower risk of recurrence (stages I and Ia) or those with co-morbidities impairing future surgery, only visits yearly or when symptoms occur. All patients should have a colonoscopy before or within 6 months of initial surgery, and repeated yearly if villous or tubular adenomas >1 cm are found; otherwise repeat every 3 to 5 years. All patients having recurrences should be assessed by a multidisciplinary team in a cancer centre. PMID- 14529579 TI - Vascular endothelial growth factor, capillarization, and function of the rat plantaris muscle at the onset of hypertrophy. AB - Capillary proliferation occurs during compensatory hypertrophy. We investigated whether the expression of vascular endothelial growth factor (VEGF) is elevated at the onset of hypertrophy when capillary proliferation is minimal, and whether muscle damage as assessed by muscle force deficits, may occur at the onset of hypertrophy. To investigate this, we induced in 9-month-old rats, under isoflurane anesthesia, hypertrophy of the left plantaris muscle by denervation of the gastrocnemius and soleus muscles. Capillarization was investigated in both the deep (oxidative) and the superficial (glycolytic) regions of the plantaris muscle. After 2 weeks, muscle mass had increased by 16% (p< 0.01), which was not accompanied by increases in fiber size. The maximal tetanic force (P(0)) and specific tension (P(0).g(-1) or P(0).cm(2)) and twitch characteristics were unaltered, and fatigue resistance of the overloaded muscle was improved (p< 0.05). However, the myosin heavy chain composition was unaltered. Capillary proliferation was not yet evident, but VEGF mRNA and protein levels were elevated 1.5- and 8-fold, respectively (p< 0.05). We concluded that the normal specific tension and the elevated VEGF expression after 2 weeks of overload indicate (1) an absence of or minimal muscle damage at this early time point, and (2) that elevated VEGF expression precedes and is involved in capillary proliferation that occurs during the later stages of compensatory hypertrophy. PMID- 14529581 TI - Repeated immobilization stress increases uncoupling protein 1 expression and activity in Wistar rats. AB - Repeat immobilization-stressed rats are leaner and have improved cold tolerance due to enhancement of brown adipose tissue (BAT) thermogenesis. This process likely involves stress-induced sympathetic nervous system activation and adrenocortical hormone release, which dynamically enhances and suppresses uncoupling protein 1 (UCP1) function, respectively. To investigate whether repeated immobilization influences UCP1 thermogenic properties, we assessed UCP1 mRNA, protein expression, and activity (GDP binding) in BAT from immobilization naive or repeatedly immobilized rats (3 h daily for 4 weeks) and sham operated or adrenalectomized (ADX) rats. UCP1 properties were assessed before (basal) and after exposure to 3 h of acute immobilization. Basal levels of GDP binding and UCP1 expression was significantly increased (140 and 140%) in the repeated immobilized group. Acute immobilization increased GDP binding in both naive (180%) and repeated immobilized groups (220%) without changing UCP1 expression. In ADX rats, basal GDP binding and UCP1 gene expression significantly increased (140 and 110%), and acute immobilization induced further increase. These data demonstrate that repeated immobilization resulted in enhanced UCP1 function, suggesting that enhanced BAT thermogenesis contributes to lower body weight gain through excess energy loss and an improved ability to maintain body temperature during cold exposure. PMID- 14529580 TI - Estrogen-induced augmentation of endothelium-dependent nitric oxide-mediated vasodilation in isolated rat cerebral small arteries. AB - We examined chronic effects of 17beta-estradiol (E(2)beta) on the responses of isolated rat anterior cerebral small arteries to vasoactive substances with special reference to endothelial function. Female Sprague-Dawley rats were separated into four groups: (1) sham-operated group (Sham), (2) sham-operated plus E(2)beta treated group (Sham+E), (3) ovariectomized group (OVX), (4) ovariectomized plus E(2)beta treated group (OVX+E). 5-Hydroxytryptamine (5-HT) (10(-10)-10(-3) M) and U46619 (10(-15)-10(-8) M) induced concentration-dependent contractions in the cerebral small arteries. The 5-HT- and U46619-induced contractions were not affected by pretreatment with 3 x 10(-5) M N(omega)-nitro-L arginine methyl ester (L-NAME). No significant difference in high potassium (80 mM)- and the agonists-mediated contractions was observed among the four groups. Administration of acetylcholine (ACh) (10(-9)-10(-3) M) and sodium nitroprusside (SNP) (10(-8)-10(-3) M) caused dose-related relaxations in the cerebral small arteries precontracted by 10(-8) M U46619. Chronic treatment with E(2)beta caused a significant potentiation of the ACh-induced relaxations in the Sham+E and OVX+E groups. The dose-response curve for ACh in the OVX group was quite similar to that obtained with the Sham group. The ACh-induced relaxation was reduced significantly by pretreatment with 3 x 10(-5) M L-NAME, and an additional treatment with 10(-3) M L-arginine reversed significantly the L-NAME-induced inhibition. The removal of endothelial cells produced a significant reduction of the ACh-induced relaxation. Indomethacin (10(-5) M) did not alter the ACh-induced relaxation. The findings suggest that E(2)beta potentiates ACh-induced endothelium-dependent relaxation in rat anterior cerebral arteries and that the potentiation may be, in part, mediated by increasing production and release of endogenous NO from the endothelial cells. PMID- 14529582 TI - Heat stress modifies human baroreflex function independently of heat-induced hypovolemia. AB - Since human thermoregulatory heat loss responses, cutaneous vasodilation and sweating, cause hypovolemia, they should resultantly stimulate human baroreflexes. However, it is possible that the thermoregulatory system directly interacts with the baroreflex system through central neural connections independently of the heat-induced hypovolemia. We hypothesized that heat stress modifies the baroreflex control of sympathetic nerve activity independently of heat-induced hypovolemia in humans. We made whole-body heating with tube-lined suits perfused with warm water (46-47 degrees C) on 10 healthy male subjects. The heating increased skin and tympanic temperatures by 10.0 and 0.4 degrees C, respectively. It increased resting total muscle sympathetic nerve activity (MSNA, microneurography) by 94 +/- 9% and decreased central venous pressure (CVP, dependent arm technique) by 2.6 +/- 0.9 mmHg. The heating increased arterial baroreflex gain by 193%, assessed as a response of MSNA to a decrease in diastolic arterial pressure during Valsalva's maneuver, but it did not change threshold arterial pressure for MSNA activation. Although the heating did not change the cardiopulmonary baroreflex gain assessed as a response of MSNA to a change in estimated central venous pressure (CVP) during a 10 degrees head-down and -up tilt test, it upwardly shifted the stimulus-response baroreflex relationship. These changes in baroreflex functions during heating were not restored by an intravenous infusion of warmed isotonic saline (37 degrees C, 15 ml/kg) that restored the heat-induced reduction of CVP. Our results support our hypothesis that heat stress modifies the baroreflex control of MSNA independently of heat-induced hypovolemia in humans. Our results also suggest that the hyperthermal modification of baroreflex results from central neural interaction between thermoregulatory and baroreflex systems. PMID- 14529583 TI - Cerebral circulation during acute microgravity induced by free drop in anesthetized rats. AB - To evaluate changes in the cerebral circulation during acute microgravity (microG), we measured intracranial pressure (ICP), aortic pressure at the diaphragm level, and cerebral flow velocity (CFV) in anesthetized rats (n = 5) during 4.5 s of microG induced by free drop, then calculated arterial pressure at the eye level (AP(eye)) and cerebral perfusion pressure (CPP = AP(eye)-ICP), and estimated CPP-CFV relationship. The rats were placed in the flat and the 30 degrees head-up positions. In the head-up position, ICP, AP(eye), and CPP were significantly increased by 2.2 +/- 0.4, 12.3 +/- 2.0, and 10.1 +/- 1.7 mmHg respectively during microG, whereas the CFV did not change significantly. In the flat position, none of these variables were significantly affected by microG. The slope of the CPP-CFV relationship was decreased only in the head-up position, suggesting that the cerebrovascular resistance was increased by microG. These findings indicate that the change in gravitational (hydrostatic) pressure is a key factor in understanding the changes in cerebral circulation during acute microG. PMID- 14529584 TI - Beta adrenoceptor agonists, clenbuterol, and isoproterenol retard denervation atrophy in rat gastrocnemius muscle: use of 3-methylhistidine as a marker of myofibrillar degeneration. AB - The effects of beta adrenergic agonists, clenbuterol (2 mg/kg body weight/d) and isoproterenol (12 mg/kg body weight/d), in normal innervated and denervated rat gastrocnemius muscle were investigated. The daily administration of beta adrenergic agonists to normal innervated rats for a short period (7 d) resulted in the hypertrophy of gastrocnemius as confirmed from the measurement of total tissue protein contents. The development of denervation atrophy witnessed a stimulation in the expression of acid and alkaline phosphatases, pointing to an enhanced myofibrillar degeneration. An administration of beta adrenergic agonists inhibited the expression of raised levels of these enzymes in denervated muscle. A measurement of 3-methylhistidine in muscle revealed a loss of amino acid with the progress in the development of denervation atrophy. Serum and urine samples from denervated rats showed a progressive accumulation of 3-methylhistidine. Clenbuterol and isoproterenol treatment to these rats resulted in an inhibition of 3-methylhistidine accumulation. When 3-methylhistidine was used as a marker of myofibrillar degeneration, the results seemed to suggest that the degeneration of cyto-contractile apparatus accompanying denervation atrophy is attenuated in the presence of beta adrenergic agonists, implying that these sympathomimetic drugs are capable of reversing denervation atrophy in rat gastrocnemius. PMID- 14529585 TI - The effects of low-intensity single-leg exercise on regional arterial stiffness. AB - We examined the effect of low-intensity single-leg exercise (20 or 30 watt, 5 min) on pulse wave velocity from the femoral to the ankle arteries in 18 young men. After the exercise, the velocity significantly decreased in the exercised leg, but not in the counterpart, suggesting that the decrease in arterial stiffness in the exercised leg was induced mainly by exercise-related regional factors. PMID- 14529586 TI - The activation of muscle spindles enhances the thixotropic behavior of rib cage respiratory muscles. AB - Rib cage inspiratory muscle contractions at a deep inflated position caused a subsequent increase in rib cage cross-sectional area at the end-expiratory phase. The increase in this area was enhanced by the activation of muscle spindles in the inspiratory muscles by a mechanical vibration combined with muscle contractions. PMID- 14529587 TI - Possible involvement of apoptotic death of myocytes in left ventricular remodeling after myocardial infarction. AB - Pathophysiological roles of apoptosis in post-infarction left ventricular (LV) remodeling have not been well characterized. This study showed that TUNEL- or cleaved caspase-3-positive myocytes were identified late after ligation of the left coronary artery in rats, suggesting that apoptotic myocyte death contributed to the morphological change associated with LV remodeling. PMID- 14529589 TI - Escaping guideline gridlock. PMID- 14529588 TI - Associations between cigarette smoking and the risk of four leading cancers in Miyagi Prefecture, Japan: a multi-site case-control study. AB - Although cigarette smoking is a well-known risk factor of lung cancer, associations of cigarette smoking with the risk of other sites have not been fully elucidated in Japan. To simultaneously evaluate the associations of cigarette smoking with the risks of cancers of the stomach, lung, colon, and rectum, which have been the leading cancer sites in recent years in Miyagi Prefecture, Japan, we conducted a hospital-based case-control study. Study subjects consisted of 614 stomach, 515 lung, 324 colon, and 164 rectal cancer cases and 2444 hospital controls admitted to a single hospital in Miyagi Prefecture from 1997 to 2001. Information on smoking habit and other lifestyle factors was collected using a self-administered questionnaire. Distributions of referral base among cases and controls were also investigated. For each site, odds ratios (ORs) and 95% confidence intervals (95% CIs) for smoking habit were estimated with adjustment for age, year of survey, history of alcohol drinking, family history of index cancer, and occupational history, respectively, using an unconditional logistic regression model. Cigarette smoking (ever vs. never) was associated with an increased risk of stomach (OR = 1.62; 95% CI 1.20-2.19) and lung (OR = 3.82; 95% CI 2.49-5.86) cancer among males and lung cancer among females (OR = 2.02; 95% CI 1.28-3.18). For female stomach cancer, the association with cigarette smoking was uncertain (OR = 0.65, P = 0.1533). For rectal cancer, a significant increased risk was observed in both-sex-combined analysis. There was no association between cigarette smoking and the risk of colon cancer. Detailed analysis showed that the association of cigarette smoking with cancer risk might be modified by the patient referral pattern, i.e., screened or not screened. The present results indicate that the association of cigarette smoking with cancer risk may differ among sites and sexes. In terms of the population attributable risk, a large proportion of leading cancers in males appears to be related to cigarette smoking. PMID- 14529590 TI - Neuraminidase inhibitors slightly beneficial for shortening flu symptoms. PMID- 14529591 TI - Tapering inhaled steroids effective for chronic asthma. PMID- 14529592 TI - Antioxidant vitamins do not prevent cardiovascular disease. PMID- 14529593 TI - Hawthorn extract improves chronic heart failure. PMID- 14529594 TI - Estrogen plus progestin may increase incidence of dementia. PMID- 14529595 TI - Thrombolytic therapy for acute ischemic stroke: risks vs benefits. PMID- 14529596 TI - Negative antigen test misses <5% of strep pharyngitis. PMID- 14529597 TI - Oral prednisone prevents relapse in COPD exacerbations. PMID- 14529598 TI - Educational interventions improve outcomes for children with asthma. PMID- 14529599 TI - Ginkgo ineffective for tinnitus. PMID- 14529600 TI - Evaluating idiopathic venous thromboembolism: what is necessary, what is not. PMID- 14529601 TI - Painful, swollen lower legs. PMID- 14529602 TI - How should we manage an acute exacerbation of COPD? AB - Would you order a chest film to evaluate an acute exacerbation of chronic obstructive pulmonary disease (COPD)? Which medication would you first prescribe a short-acting inhaled beta-2 agonist or an anticholinergic bronchodilator? These are important questions for family physicians who commonly manage acute exacerbations of COPD. The guideline summarized here was developed by a joint expert panel of the American College of Physicians-American Society of Internal Medicine and the American College of Chest Physicians. Three outcomes were considered: treatment efficacy, 6-month mortality, and relapse, as defined by return visit to the emergency department within 14 days of initial presentation. Systematic reviews with evidence tables were used to analyze data. The rationale for each recommendation is clear and well documented. We added strength-of recommendation ratings, which are not in the original guideline. PMID- 14529603 TI - How practice-based research changed the way I manage depression. PMID- 14529604 TI - Jimmy Holyfield's last joke. PMID- 14529605 TI - Tuberculosis: old problem, new concerns. PMID- 14529606 TI - Clinical inquiries. What regimens eradicate Helicobacter pylori? PMID- 14529607 TI - Clinical inquiries. Does a knee brace decrease recurrent ACL injuries? PMID- 14529608 TI - Clinical inquiries. Does breastfeeding protect against viral GI infections in children <2 years old? PMID- 14529609 TI - Clinical inquiries. Which infants need lumbar puncture for suspected sepsis? PMID- 14529610 TI - Clinical inquiries. What medication best prevents migraine in children? PMID- 14529612 TI - Interaction of the MAGUK family member Acvrinp1 and the cytoplasmic domain of the Notch ligand Delta1. AB - The evolutionarily conserved Notch signal transduction pathway regulates cell fate and cellular differentiation in various tissues and has essential functions in embryonic patterning and tumorigenesis. Cell-cell signaling by the Notch pathway is mediated by the interaction of the transmembrane receptor Notch with its ligands Delta or Jagged presented on adjacent cells. Whereas signal transduction to Notch expressing cells has been described, it is unclear whether Delta-dependent signaling may exist within the Delta-expressing cell. Here, we report on the identification of Acvrinp1, a MAGUK family member, interacting with the intracellular domain of Delta1 (Dll1). We confirmed the interaction between Dll1 and Acvrinp1 by pull-down experiments in vitro and in a mammalian two-hybrid system in vivo. We delimited the fourth PDZ domain of Acvrinp1 and the PDZ binding domain of Dll1 as major interacting domains. In situ expression analyses in mouse embryos revealed that Dll1 and Acvrinp1 show partly overlapping but distinct expression patterns, for example, in the central nervous system and the vibrissae buds. Further, we found that expression of Acvrinp1 is altered in Dll1 loss-of-function mouse embryos. PMID- 14529613 TI - Neuron cell type-specific SNAP-25 expression driven by multiple regulatory elements in the nematode Caenorhabditis elegans. AB - In order to characterize the mechanisms regulating neuronal expression of the nematode SNAP-25 gene, we identified the SNAP-25 genes of Caenorhabditis elegans and Caenorhabditis briggsae. Comparative sequence analysis and reporter assays revealed two putative 5' regulatory elements, P1 and P2, and four elements, I1h, I1m, I2h, and I2m, in the first intron. Nuclear extracts contained activities that bound the P2 and I1h elements. Different elements were required for SNAP-25 expression in different neuronal subsets; P1 was required in DA and DD motor neurons, and I1m and I2m were required in DB and DA neurons, respectively. P2 was active in amphid and phasmid neurons, I1h in pharyngeal neurons, and I2h in touch receptor neurons. The I2h element contained a putative binding site for transcription factor UNC-86. Both UNC-86 and MEC-3 were required for I2h activity in the mechanosensory neurons: in these neurons, GFP expression driven by I2h was abolished in animals bearing either an unc-86 null or a mec-3 null mutation, or an unc-86 mutation that leads to defective interaction with MEC-3. Deletion of the MEC-3 binding site also abolished the GFP expression. Gel mobility-shift assay results suggest that transcriptional regulation of SNAP-25 may involve multiple transcription factors. PMID- 14529614 TI - Proteasome inhibition induces nuclear translocation of the dioxin receptor through an Sp1 and protein kinase C-dependent pathway. AB - The dioxin receptor (AhR), in addition to its role in xenobiotic-induced carcinogenesis, appears to participate in cell proliferation, differentiation and organ homeostasis. Understanding potential mechanisms of activation of this receptor in the absence of exogenous ligands is therefore important to study its contribution to endogenous cellular functions. Using mouse embryo primary fibroblasts, we have previously shown that proteasome inhibition increased AhR transcriptional activity in the absence of xenobiotics. We suggested that proteasome inhibition-dependent AhR activation could involve an increase in the expression of the partner protein dioxin receptor nuclear translocator (ARNT). Since ARNT over-expression induced nuclear translocation of the AhR, and ARNT deficient cells were unable to translocate this receptor to the nucleus upon proteasome inhibition, we have analyzed the effect of proteasome inhibition on the expression of regulatory proteins controlling ARNT levels. Treatment with the proteasome inhibitor MG132 increased endogenous Sp1 phosphorylation and its DNA binding activity to the ARNT promoter. Sp1 phosphorylation and binding to the ARNT promoter, ARNT over-expression and AhR nuclear translocation were inhibited by GF109203X, a protein kinase C-specific inhibitor. In addition, MG132 stimulated protein kinase C activity in MEF cells with a pattern similar to that observed for ARNT expression. These data suggest that cellular control of protein kinase C activity, through Sp1 and ARNT, could regulate AhR transcriptional activity in the absence of xenobiotics. PMID- 14529615 TI - Sigma70 promoters in Escherichia coli: specific transcription in dense regions of overlapping promoter-like signals. AB - We present here a computational analysis showing that sigma70 house-keeping promoters are located within zones with high densities of promoter-like signals in Escherichia coli, and we introduce strategies that allow for the correct computer prediction of sigma70 promoters. Based on 599 experimentally verified promoters of E.coli K-12, we generated and evaluated more than 200 weight matrices optimizing different criteria to obtain the best recognition matrices. The alignments generating the best statistical models did not fully correspond with the canonical sigma70 model. However, matrices that correspond to such a canonical model performed better as tools for prediction. We tested the predictive capacity of these matrices on 250 bp long regions upstream of gene starts, where 90% of the known promoters occur. The computational matrix models generated an average of 38 promoter-like signals within each 250 bp region. In more than 50% of the cases, the true promoter does not have the best score within the region. We observed, in fact, that real promoters occur mostly within regions with high densities of overlapping putative promoters. We evaluated several strategies to identify promoters. The best one uses an intrinsic score of the -10 and -35 hexamers that form the promoter as well as an extrinsic score that uses the distribution of promoters from the start of the gene. We were able to identify 86% true promoters correctly, generating an average of 4.7 putative promoters per region as output, of which 3.7, on average, exist in clusters, as a series of overlapping potentially competing RNA polymerase-binding sites. As far as we know, this is the highest predictive capability reported so far. This high signal density is found mainly within regions upstream of genes, contrasting with coding regions and regions located between convergently transcribed genes. These results are consistent with experimental evidence that show the existence of multiple overlapping promoter sites that become functional under particular conditions. This density is probably the consequence of a rich number of vestiges of promoters in evolution. We suggest that transcriptional regulators as well as other functional promoters play an important role in keeping these latent signals suppressed. PMID- 14529616 TI - The mechanism of proton exclusion in the aquaporin-1 water channel. AB - Aquaporins are efficient, yet strictly selective water channels. Remarkably, proton permeation is fully blocked, in contrast to most other water-filled pores which are known to conduct protons well. Blocking of protons by aquaporins is essential to maintain the electrochemical gradient across cellular and subcellular membranes. We studied the mechanism of proton exclusion in aquaporin 1 by multiple non-equilibrium molecular dynamics simulations that also allow proton transfer reactions. From the simulations, an effective free energy profile for the proton motion along the channel was determined with a maximum-likelihood approach. The results indicate that the main barrier is not, as had previously been speculated, caused by the interruption of the hydrogen-bonded water chain, but rather by an electrostatic field centered around the fingerprint Asn-Pro-Ala (NPA) motif. Hydrogen bond interruption only forms a secondary barrier located at the ar/R constriction region. The calculated main barrier height of 25-30 kJ mol( 1) matches the barrier height for the passage of protons across pure lipid bilayers and, therefore, suffices to prevent major leakage of protons through aquaporins. Conventional molecular dynamics simulations additionally showed that negatively charged hydroxide ions are prevented from being trapped within the NPA region by two adjacent electrostatic barriers of opposite polarity. PMID- 14529617 TI - Functional requirement for symmetric assembly of archaeal box C/D small ribonucleoprotein particles. AB - Box C/D small ribonucleoprotein particles (sRNPs) are archaeal homologs of small nucleolar ribonucleoprotein particles (snoRNPs) in eukaryotes that are responsible for site specific 2'-O-methylation of ribosomal and transfer RNAs. The function of box C/D sRNPs is characterized by step-wise assembly of three core proteins around a box C/D RNA that include fibrillarin, Nop5p, and L7Ae. The most distinct structural feature in all box C/D RNAs is the presence of two conserved box C/D motifs accompanied by often a single, and sometimes two, antisense elements located immediately upstream of either the D or D' box. Despite this asymmetric distribution of antisense elements, the bipartite feature of the box C/D motifs appears to be in pleasing agreement with a recently reported three-dimensional structure of the core protein complex between fibrillarin and Nop5p. This investigates functional implications of the symmetric features both in box C/D RNAs and in the fibrillarin-Nop5p complex. Site-directed mutagenesis was employed to generate box C/D RNAs lacking one of the two box C/D motifs and a mutant fibrillarin-Nop5p complex deficient in self-association. The ability of the mutated components to assemble and to direct methyl transfer reactions was assessed by gel mobility-shift, analytical ultracentrifugation, and in vitro catalysis studies. The results presented here suggest that, while a box C/D sRNP is capable of asymmetrical assembly, the symmetries in both the box C/D RNA and in the fibrillarin-Nop5p complex are required for efficient catalysis. These findings underscore the importance of functional assembly in methyl transfer reactions. PMID- 14529618 TI - In vivo and in vitro evidence that the four essential intermediate filament (IF) proteins A1, A2, A3 and B1 of the nematode Caenorhabditis elegans form an obligate heteropolymeric IF system. AB - The in vitro polymerization and tissue-specific expression patterns of the four essential intermediate filament (IF) proteins (A1, A2, A3, B1) and the non essential IF protein A4 were analyzed. Recombinant B1, used as a probe in blot overlay assays of the 11 Caenorhabditis elegans IF proteins, reacted strongly with proteins A1 to A4, indicating a heterotypic interaction. Obligate heteropolymeric filament assembly in vitro was confirmed by electron microscopy. Protein B1 formed long IF when mixed with an equimolar amount of A1, A2 or A3. Developmentally regulated coexpression of B1 and one or more members of the A family was found with GFP-promoter reporters. This coexpression pattern argues for a heteropolymer system in vivo. One or both splice variants of the B1 gene are always coexpressed in a tissue-specific manner with at least one member of the A family in hypodermis, pharynx, pharyngeal-intestinal valve, excretory cells, uterus, vulva and rectum. Interestingly, while the intestine normally lacks a B1/A pair, the dauer larva shows intestinal B1 and A4. These results are in line with similar postembryonic phenotypes of the hypodermis induced by RNA interference (RNAi) of genes B1, A2 and A3. Similarly, defects of the pharynx and its A1-GFP containing tonofilaments observed in the postembryonic B1 RNAi phenotype are consistent with the coexpression of B1 and A1 in the marginal cells. Thus RNAi analyses provide independent evidence for the existence of the B1/A obligate heteropolymer system in vivo. Proteins A1 and B1 have a similar and rather slow turnover rate in photobleaching experiments of the pharynx tonofilaments. PMID- 14529619 TI - S-adenosyl methionine prevents promiscuous DNA cleavage by the EcoP1I type III restriction enzyme. AB - DNA cleavage by the type III restriction endonuclease EcoP1I was analysed on circular and catenane DNA in a variety of buffers with different salts. In the presence of the cofactor S-adenosyl methionine (AdoMet), and irrespective of buffer, only substrates with two EcoP1I sites in inverted repeat were susceptible to cleavage. Maximal activity was achieved at a Res2Mod2 to site ratio of approximately 1:1 yet resulted in cleavage at only one of the two sites. In contrast, the outcome of reactions in the absence of AdoMet was dependent upon the identity of the monovalent buffer components, in particular the identity of the cation. With Na+, cleavage was observed only on substrates with two sites in inverted repeat at elevated enzyme to site ratios (>15:1). However, with K+ every substrate tested was susceptible to cleavage above an enzyme to site ratio of approximately 3:1, including a DNA molecule with two directly repeated sites and even a DNA molecule with a single site. Above an enzyme to site ratio of 2:1, substrates with two sites in inverted repeat were cleaved at both cognate sites. The rates of cleavage suggested two separate events: a fast primary reaction for the first cleavage of a pair of inverted sites; and an order-of-magnitude slower secondary reaction for the second cleavage of the pair or for the first cleavage of all other site combinations. EcoP1I enzymes mutated in either the ATPase or nuclease motifs did not produce the secondary cleavage reactions. Thus, AdoMet appears to play a dual role in type III endonuclease reactions: Firstly, as an allosteric activator, promoting DNA association; and secondly, as a "specificity factor", ensuring that cleavage occurs only when two endonucleases bind two recognition sites in a designated orientation. However, given the right conditions, AdoMet is not strictly required for DNA cleavage by a type III enzyme. PMID- 14529620 TI - Fourteen protomers compose the oligomer III of the proton-rotor in spinach chloroplast ATP synthase. AB - Three fundamentally different chloroplast ATP synthase samples of increasing complexity were visualized by atomic force microscopy. The samples are distinguishable in respect to the isolation technique, the detergent employed, and the final subunit composition. The homo-oligomer III was isolated following SDS treatment of ATP synthase, the proton-turbine III+IV was obtained by blue native electrophoresis, and complete CFO was isolated by anion exchange chromatography of NaSCN splitted ATP synthase. In all three ATP synthase subcomplexes 14 and only 14 circularly arranged subunits III composed the intact transmembrane rotor. Therefore, 14 protomers built the membrane-resident proton turbine. The observed stoichiometry of 14 is not a biochemical artifact or affected by natural growth variations of the spinach, as previously suggested. A correlation between the presence of subunit IV in the imaged sample and the appearance of a central protrusion in the narrower orifice of the oligomeric cylinder III14 has been observed. In contrast to current predictions, in chloroplast FO the subunit IV can be found inside the cylinder III14 and not at its periphery, at least in the reconstituted 2D arrays imaged. PMID- 14529621 TI - Complexes of RecA with LexA and RecX differentiate between active and inactive RecA nucleoprotein filaments. AB - The bacterial RecA protein has been the dominant model system for understanding homologous genetic recombination. Although a crystal structure of RecA was solved ten years ago, we still do not have a detailed understanding of how the helical filament formed by RecA on DNA catalyzes the recognition of homology and the exchange of strands between two DNA molecules. Recent structural and spectroscopic studies have suggested that subunits in the helical filament formed in the RecA crystal are rotated when compared to the active RecA-ATP-DNA filament. We examine RecA-DNA-ATP filaments complexed with LexA and RecX to shed more light on the active RecA filament. The LexA repressor and RecX, an inhibitor of RecA, both bind within the deep helical groove of the RecA filament. Residues on RecA that interact with LexA cannot be explained by the crystal filament, but can be properly positioned in an existing model for the active filament. We show that the strand exchange activity of RecA, which can be inhibited when RecX is present at very low stoichiometry, is due to RecX forming a block across the deep helical groove of the RecA filament, where strand exchange occurs. It has previously been shown that changes in the nucleotide bound to RecA are associated with large motions of RecA's C-terminal domain. Since RecX binds from the C terminal domain of one subunit to the nucleotide-binding core of another subunit, a stabilization of RecA's C-terminal domain by RecX can likely explain the inhibition of RecA's ATPase activity by RecX. PMID- 14529622 TI - Binders based on dimerised immunoglobulin VH domains. AB - Antibody binding to antigen is mediated by the surface formed by the association of the two variable (V) regions of the L (VL) and H (VH) chains. The capacity of VL to dimerise and the high structural similarity of VL and VH domains suggested the possibility that VH could also associate. We show here that spontaneous formation of VH dimers (VHD) is in many cases permissive, producing stable molecules with antigen binding specificity. VHD were displayed on filamentous phages for the selection of antigen-specific binders. VHD were expressed and secreted efficiently from both bacteria and mammalian cells in different formats, including single-chain (VH(1)-linker-VH(2)), double chain ((VH(2)) and IgG analogues having the VL replaced by VH. The affinity (Kd,app) achieved with a VH dimer expressed in the IgG format, specific for a glutenin subunit was around 30 nM measured by two different methods, which was about 20 times higher than that corresponding to the VL/VH counterpart. PMID- 14529623 TI - Crystal structures of the free and anisic acid bound triple mutant of phospholipase A2. AB - Phospholipase A2 catalyses the hydrolysis of the ester bond of 3-sn phosphoglycerides. Here, we report the crystal structures of the free and anisic acid-bound triple mutant (K53,56,120M) of bovine pancreatic phospholipase A2. In the bound triple mutant structure, the small organic molecule p-anisic acid is found in the active site, and one of the carboxylate oxygen atoms is coordinated to the functionally important primary calcium ion. The other carboxylate oxygen atom is hydrogen bonded to the phenolic hydroxyl group of Tyr69. In addition, the bound anisic acid molecule replaces one of the functionally important water molecules in the active site. The residues 60-70, which are in a loop (surface loop), are disordered in most of the bovine pancreatic phospholipase A2 structures. It is interesting to note that these residues are ordered in the bound triple mutant structure but are disordered in the free triple mutant structure. The organic crystallization ingredient 2-methyl-2,4-pentanediol is found near the active site of the free triple mutant structure. The overall tertiary folding and stereochemical parameters for the final models of the free and anisic acid-bound triple mutant are virtually identical. PMID- 14529624 TI - Crystal structure of putidaredoxin, the [2Fe-2S] component of the P450cam monooxygenase system from Pseudomonas putida. AB - Stability of the [2Fe-2S]-containing putidaredoxin (Pdx), the electron donor to cytochrome P450cam in Pseudomonas putida, was improved by mutating non-ligating cysteine residues, Cys73 and Cys85, to serine singly and in combination. The increasing order of stability is Cys73Ser/Cys85Ser>Cys73Ser>Cys85Ser>WT Pdx. Crystal structures of Cys73Ser/Cys85Ser and Cys73Ser mutants of Pdx, solved by single-wavelength anomalous dispersion phasing using the [2Fe-2S] iron atoms to 1.47 A and 1.65 A resolution, respectively, are nearly identical and very similar to those of bovine adrenodoxin (Adx) and Escherichia coli ferredoxin. However, unlike the Adx structure, no motion between the core and interaction domains of Pdx is observed. This higher conformational stability of Pdx might be due to the presence of a more extensive hydrogen bonding network at the interface between the two structural domains around the conserved His49. In particular, formation of a hydrogen bond between the side-chain of Tyr51 and the carbonyl oxygen atom of Glu77 and the presence of two well-ordered water molecules linking the interaction domain and the C-terminal peptide to the core of the molecule are unique to Pdx. The folding topology of the NMR model is similar to that of the X ray structure of Pdx. The overall rmsd of Calpha positions between the two models is 1.59 A. The largest positional differences are observed for residues 18-21 and 33-37 in the loop regions and the C terminus. The latter two peptides display conformational heterogeneity in the crystal structures. Owing to flexibility, the aromatic ring of the C-terminal Trp106 can closely approach the side-chains of Asp38 and Thr47 (3.2-3.9 A) or move away and leave the active site solvent exposed. Therefore, Trp106, previously shown to be important in the Pdr-to-Pdx and Pdx-to-P450cam electron transfer reactions is in a position to regulate and/or mediate electron transfer to or from the [2Fe-2S] center of Pdx. PMID- 14529625 TI - Structural characterization of the GSK-3beta active site using selective and non selective ATP-mimetic inhibitors. AB - GSK-3beta is a regulatory serine/threonine kinase with a plethora of cellular targets. Consequently, selective small molecule inhibitors of GSK-3beta may have a variety of therapeutic uses including the treatment of neurodegenerative diseases, type II diabetes and cancer. In order to characterize the active site of GSK-3beta, we determined crystal structures of unphosphorylated GSK-3beta in complex with selective and non-selective ATP-mimetic inhibitors. Analysis of the inhibitors' interactions with GSK-3beta in the structures reveals how the enzyme can accommodate a number of diverse molecular scaffolds. In addition, a conserved water molecule near Thr138 is identified that can serve a functional role in inhibitor binding. Finally, a comparison of the interactions made by selective and non-selective inhibitors highlights residues on the edge of the ATP binding site that can be used to obtain inhibitor selectivity. Information gained from these structures provides a promising route for the design of second-generation GSK-3beta inhibitors. PMID- 14529627 TI - Identification of a residue critical for maintaining the functional conformation of BPTI. AB - The effects of amino acid replacements on the backbone dynamics of bovine pancreatic trypsin inhibitor (BPTI) were examined using 15N NMR relaxation experiments. Previous studies have shown that backbone amide groups within the trypsin-binding region of the wild-type protein undergo conformational exchange processes on the micros time scale, and that replacement of Tyr35 with Gly greatly increases the number of backbone atoms involved in such motions. In order to determine whether these mutational effects are specific to the replacement of this residue with Gly, six additional replacements were examined in the present study. In two of these, Tyr35 was replaced with either Ala or Leu, and the other four were single replacements of Tyr23, Phe33, Asn43 or Asn44, all of which are highly buried in the native structure and conserved in homologous proteins. The Y35A and Y35L mutants displayed dynamic properties very similar to those of the Y35G mutant, with the backbone segments including residues 10-19 and 32-44 undergoing motions revealed by enhanced 15N transverse relaxation rates. On the other hand, the Y23L, N43G and N44A substitutions caused almost no detectable changes in backbone dynamics, on either the ns-ps or ms-micros time scales, even though each of these replacements significantly destabilizes the native conformation. Replacement of Phe33 with Leu caused intermediate effects, with several residues that have previously been implicated in motions in the wild-type protein displaying enhanced transverse relaxation rates. These results demonstrate that destabilizing amino acid replacements can be accommodated in a native protein with dramatically different effects on conformational dynamics and that Tyr35 plays a particularly important role in defining the conformation of the trypsin-binding site of BPTI. PMID- 14529626 TI - Three-dimensional structure of the channel-forming trans-membrane domain of virus protein "u" (Vpu) from HIV-1. AB - The three-dimensional structure of the channel-forming trans-membrane domain of virus protein "u" (Vpu) of HIV-1 was determined by NMR spectroscopy in micelle and bilayer samples. Vpu(2-30+) is a 36-residue polypeptide that consists of residues 2-30 from the N terminus of Vpu and a six-residue "solubility tag" at its C terminus that facilitates the isolation, purification, and sample preparation of this highly hydrophobic minimal channel-forming domain. Nearly all of the resonances in the two-dimensional 1H/15N HSQC spectrum of uniformly 15N labeled Vpu(2-30+) in micelles are superimposable on those from the corresponding residues in the spectrum of full-length Vpu, which indicates that the structure of the trans-membrane domain is not strongly affected by the presence of the cytoplasmic domain at its C terminus. The two-dimensional 1H/15N PISEMA spectrum of Vpu(2-30+) in lipid bilayers aligned between glass plates has been fully resolved and assigned. The "wheel-like" pattern of resonances in the spectrum is characteristic of a slightly tilted membrane-spanning helix. Experiments were also performed on weakly aligned micelle samples to measure residual dipolar couplings and chemical shift anisotropies. The analysis of the PISA wheels and Dipolar Waves obtained from both weakly and completely aligned samples show that Vpu(2-30+) has a trans-membrane alpha-helix spanning residues 8-25 with an average tilt of 13 degrees. The helix is kinked slightly at Ile17, which results in tilts of 12 degrees for residues 8-16 and 15 degrees for residues 17-25. A structural fit to the experimental solid-state NMR data results in a three dimensional structure with precision equivalent to an RMSD of 0.4 A. Vpu(2-30+) exists mainly as an oligomer on PFO-PAGE and forms ion-channels, a most frequent conductance of 96(+/- 6) pS in lipid bilayers. The structural features of the trans-membrane domain are determinants of the ion-channel activity that may be associated with the protein's role in facilitating the budding of new virus particles from infected cells. PMID- 14529628 TI - Conversion of wild-type p53 core domain into a conformation that mimics a hot spot mutant. AB - The wild-type p53 protein can be driven into a conformation corresponding to that adopted by structural mutant forms by heterodimerization with a mutant subunit. To seek partially folded states of the wild-type p53 core domain (p53C) we used high hydrostatic pressure (HP) and subzero temperatures. Aggregation of the protein was observed in parallel with its pressure denaturation at 25 and 37 degrees C. However, when HP experiments were performed at 4 degrees C, the extent of denaturation and aggregation was significantly less pronounced. On the other hand, subzero temperatures under pressure led to cold denaturation and yielded a non-aggregated, alternative conformation of p53C. Nuclear magnetic resonance (1H15N-NMR) data showed that the alternative p53C conformation resembled that of the hot-spot oncogenic mutant R248Q. This alternative state was as susceptible to denaturation and aggregation as the mutant R248Q when subjected to HP at 25 degrees C. Together these data demonstrate that wild-type p53C adopts an alternative conformation with a mutant-like stability, consistent with the dominant-negative effect caused by many mutants. This alternative conformation is likely related to inactive forms that appear in vivo, usually driven by interaction with mutant proteins. Therefore, it can be a valuable target in the search for ways to interfere with protein misfolding and hence to prevent tumor development. PMID- 14529629 TI - Sequence alignments and pair hidden Markov models using evolutionary history. AB - This work presents a novel pairwise statistical alignment method based on an explicit evolutionary model of insertions and deletions (indels). Indel events of any length are possible according to a geometric distribution. The geometric distribution parameter, the indel rate, and the evolutionary time are all maximum likelihood estimated from the sequences being aligned. Probability calculations are done using a pair hidden Markov model (HMM) with transition probabilities calculated from the indel parameters. Equations for the transition probabilities make the pair HMM closely approximate the specified indel model. The method provides an optimal alignment, its likelihood, the likelihood of all possible alignments, and the reliability of individual alignment regions. Human alpha and beta-hemoglobin sequences are aligned, as an illustration of the potential utility of this pair HMM approach. PMID- 14529630 TI - The Role of zinc in the disulphide stress-regulated anti-sigma factor RsrA from Streptomyces coelicolor. AB - The regulation of disulphide stress in actinomycetes such as Streptomyces coelicolor is known to involve the zinc-containing anti-sigma factor RsrA that binds and inactivates the redox-regulated sigma factor sigmaR. However, it is not known how RsrA senses disulphide stress nor what role the metal ion plays. Using in vitro assays, we show that while zinc is not required for sigmaR binding it is required for functional anti-sigma factor activity, and that it plays a critical role in modulating the reactivity of RsrA cysteine thiol groups towards oxidation. Apo-RsrA is easily oxidised and, while the Zn-bound form is relatively resistant, the metal ion is readily expelled when the protein is treated with strong oxidants such as diamide. We also show, using a combination of proteolysis and mass spectrometry, that the first critical disulphide to form in RsrA involves Cys11 and one of either Cys41 or Cys44, all previously implicated in metal binding. Circular dichroism spectroscopy was used to follow structural changes during oxidation of RsrA, which indicated that concomitant with formation of this critical disulphide bond is a major restructuring of the protein where its alpha-helical content increases. Our data demonstrate that RsrA can only bind sigmaR in the reduced state and that this state is stabilised by zinc. Redox stress induces disulphide bond formation amongst zinc-ligating residues, expelling the metal ion and stabilising a structure incapable of binding the sigma factor. PMID- 14529631 TI - Nosocomial influenza in children. AB - Influenza is the most important cause of acute respiratory illness leading to hospitalization among children during community epidemics. This illness can cause extensive nosocomial outbreaks with serious morbidity and mortality among specific groups of children. Paediatric patients with community-acquired influenza and healthcare workers are the main reservoir for the nosocomial spread of the virus. During epidemics in the community, testing for influenza should be requested in all children with compatible symptoms admitted in the hospital, and measures should be introduced for the prevention or early control of an outbreak. Recent advances in the management of influenza include rapid diagnoses based on antigen detection and the identification of the new neuraminidase inhibitors zanamivir and oseltamivir. Annual vaccination against influenza of children with high-risk conditions, their family members and healthcare workers is the principle measure for the prevention of nosocomial outbreaks. Although vaccination against influenza appears to be cost-effective at all ages in terms of prevention of illness, related hospitalizations, deaths, reduction of healthcare costs and productivity loss, vaccination coverage among target groups is limited. PMID- 14529632 TI - The addition of a mobile ultra-clean exponential laminar airflow screen to conventional operating room ventilation reduces bacterial contamination to operating box levels. AB - A mobile screen producing ultra-clean exponential laminar airflow (LAF) was investigated as an addition to conventional turbulent/mixing operating room (OR) ventilation (16 air changes/h). The evaluation was performed in a small OR (50 m(3)) during 60 standardized operations for groin hernia including mesh implantation. The additional ventilation was used in 50 of the operations. The LAF passed from the foot-end of the OR table over the instrument and surgical area. Strict hygiene OR procedures including tightly woven and non-woven OR clothing were used. Sedimentation rates were recorded at the level of the patients' chests (N=60) (i.e. the air had passed the surgical team) and in the periphery of the OR. In addition bacterial air contamination was studied above the patients' chests in all 10 operations without the additional LAF and in 12 with the LAF. The screen reduced the mean counts of sedimenting bacteria (cfu/m(2)/h) on the patients' chests from 775 without the screen to 355 (P=0.0003). The screen also reduced the mean air counts of bacteria (cfu/m(3)) above the patients' chests from 27 to 9 (P=0.0001). No significant differences in mean sedimentation rates (cfu/m(2)/h) existed in the periphery of the OR where 628 without and 574 with screen were recorded. During the follow-up period of six months no surgical site infections were detected. In conclusion when the mobile LAF screen was added to conventional OR ventilation the counts of aerobic airborne and sedimenting bacteria-carrying particles downstream of the surgical team were reduced to the levels achieved with complete ultra-clean LAF OR ventilation (operating box). PMID- 14529633 TI - Development of bacterial resistance to several biocides and effects on antibiotic susceptibility. AB - The aims of this study were to investigate the development of bacterial resistance to eugenol, thymol, trichlorocarbanalide (TCC), didecyldimethylammonium chloride (DDDMAC) and C10-16-alkyldimethyl, N-oxides (ADMAO) and subsequent effects on antibiotic susceptibility. An agar minimum inhibitory concentration (MIC) method was used to assess the activity of the biocides against standard bacterial strains and laboratory mutants. A range of techniques including disk diffusion and gradient plate experiments were used to attempt to develop bacterial 'resistance' or tolerance to the biocides. The mutants produced were examined for cross-resistance to the other biocides and to antibiotics via disk diffusion and gradient plate MIC methods. Outer membrane proteins of the mutants were extracted and examined using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Escherichia coli triclosan resistant mutants were not cross-resistant to eugenol, thymol, TCC, DDDMAC and ADMAO. Mutants with elevated MICs to DDDMAC (E. coli and Pseudomonas aeruginosa), thymol (E. coli) and eugenol (E. coli) were isolated, but all remained sensitive to higher concentrations of the agents. Bacteria with elevated MICs to TCC and ADMAO were not obtained. Some low-level cross-resistance between DDDMAC, eugenol and thymol was observed with the E. coli gradient plate mutants, as well as reduced susceptibility to antibiotics, most notably chloramphenicol. The lack of cross-resistance of the triclosan mutants suggested that the mode of action of triclosan is not shared with the other biocides studied. SDS-PAGE results indicated that the DDDMAC P. aeruginosa mutant had a reduced amount (or absence) of one outer membrane protein in comparison with the standard strain. In conclusion, under laboratory conditions, bacterial exposure to thymol, eugenol and DDDMAC can lead to reduced susceptibility between selected biocidal agents and antibiotics, more specifically, chloramphenicol. However, further studies are required to determine if this is of clinical significance. PMID- 14529634 TI - A standardized test to assess the impact of different organic challenges on the antimicrobial activity of antiseptics. AB - This paper presents a suggestion for determining the antimicrobial efficacy of antiseptics in a phase 2/step 1 assay. Emphasis is placed on specific organic challenges to evaluate the efficacy of antiseptics used to rinse the oral cavity, mucous membranes, or wounds for either prophylactic or therapeutic reasons. Basic data are given concerning ready-to-use products based on Poly(1-vinyl-2 pyrrolidone-)iodine-complex (PVP-iodine), chlorhexidine, octenidine, or cetylpyridinium chloride. Hydrogen peroxide (3.0%) was included to assess a product with known limited efficacy only. Using Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Enterococcus faecium, and Candida albicans, a 10(5)-fold reduction can be achieved in 10 min without interfering substances. In the presence of 10% albumin, 10% sheep blood, or 1% mucin, a 10(3) fold reduction is yielded using products based on PVP-iodine, octenidine, or chlorhexidine. The combination of 4.5% albumin, 4.5% sheep blood, and 1% mucin was the most difficult organic challenge and only the products based on PVP iodine, octenidine, or chlorhexidine remained active. The cetylpyridinium-based product and hydrogen peroxide were less effective. Based on these data, a proposal for a phase 2/step 1 assay for antiseptics is presented to promote the standardization efforts. PMID- 14529635 TI - Interaction between methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA). AB - In this paper we describe the in vitro interaction between three strains of methicillin-sensitive Staphylococcus aureus (MSSA) [NCTC 11561 and two strains derived from patients (PMSSA)] and endemic methicillin-resistant Staphylococcus aureus (EMRSA) 1, 3, 15 and 16. Mixed bacterial cultures of MSSA and EMRSA were incubated and subcultured after one, two and seven days. A proportion of MRSA in 50 randomly selected colonies was assessed. All strains of EMRSA (EMRSA 1, 3, 15 and 16) outgrew and virtually eradicated MSSA (NCTC 11561) after 24 h. The interaction between strains of PMSSA and the various strains of EMRSA was variable. PMSSA strain 1 was almost completely outgrown by EMRSA 1 and EMRSA 3 after seven days. Similarly there was a substantial increase of EMRSA 1 and 3 when tested against PMSSA strain 2. EMRSA 15 increased modestly against both strains, but EMRSA 16 failed to increase in proportion against either of the strains. We conclude that there is a complex interaction between various strains of EMRSA and MSSA. This interaction may have an important bearing on colonization of patients with MRSA. PMID- 14529636 TI - Prevalence of urinary catheterization in UK nursing homes. AB - A postal questionnaire survey was undertaken in registered nursing homes in three different health districts in England: Gloucestershire, North Staffordshire and Leeds. Nursing homes may be registered as general nursing or mental health homes. If homes also have provision for residential beds these are defined as dual registered homes. Overall, 9% (438/4900) of residents, with an equal male:female split, had urinary catheters. There was no significant difference in the overall urinary catheterization rate in the three districts (P=0.9). There was a wide range of urinary catheterization prevalence between homes, with some homes of all three categories having no catheterized residents and several with a prevalence of over 40%. The wide range of prevalence may be due to differences in residents' underlying medical conditions or to differences in attitudes towards urinary catheterization by nursing home staff. Almost all homes (114/124, 92%) stated they had an infection control policy, but 31% (38/124) did not have a written policy on urinary catheter care. In view of the potential for morbidity, infection control policies should include a section on the care of urinary catheters and this should form part of the continuing training of nursing home staff. PMID- 14529637 TI - Comparison of AP-PCR typing and PCR-ribotyping for estimation of nosocomial transmission of Clostridium difficile. AB - We recently attempted to clarify an increased incidence of Clostridium difficile associated diarrhoea (CDAD) in our hospital by arbitrarily primed polymerase chain reaction (AP-PCR) typing of isolates from 147 consecutive patients collected during a 12 month period (Wullt et al. J Hosp Infect 1999;43:265-273). In the present study we compared the results based on previous AP-PCR data with those based on recent PCR ribotyping of the same isolates and re-analysis of a subset of isolates by AP-PCR typing. The pattern of PCR ribotypes was similar among inpatients and outpatients. A cluster of three closely related PCR ribotypes, related to those of the serogroup H and A8 type strains, dominated and comprised 31% of inpatient and 28% of outpatient C. difficile isolates. The apparent nosocomial transmission rate among inpatients with CDAD was only 9% by AP-PCR typing compared with 18 or 36% by PCR ribotyping depending on the definition used (proportion of patients sharing C. difficile type and ward within two or 12 months). Corresponding rates for all CDAD patients were 5% by AP-PCR and 11 or 21% by PCR ribotyping. Thus, most CDAD patients apparently became ill due to their endogenous strain of C. difficile. Because of the low concordance between the two typing methods the proportion of patients fulfilling the criteria for nosocomial transmission by both methods was only 1%. Re-examination of isolates from patients with recurrences revealed a reproducibility problem with AP-PCR typing. We conclude, that of these two PCR-based options for typing of C. difficile PCR ribotyping offers a superior experimental robustness compared with AP-PCR typing. PMID- 14529638 TI - Gastroenteritis outbreak with norovirus in a Swiss university hospital with a newly identified virus strain. AB - We report an outbreak of norovirus-associated gastroenteritis in patients and healthcare workers (HCWs) at a university hospital in Switzerland during the period from 28 February to 31 March 2001. Faecal and vomitus specimens and bottled and drinking water were investigated for norovirus by reverse transcriptase-polymerase chain reaction (RT-PCR) Sixty-three patients and HCWs were affected. 37% of the investigated stool specimens were positive for norovirus. Sequencing showed a new phylogenetic strain, "Basel". There was no evidence for a water-borne, foodborne or environmental source. The source of the outbreak was most likely a patient admitted to the hospital. Once an outbreak was suspected, measures were instituted based on published guidelines, such as isolation of patients and excluding sick HCWs from work. However, the application of the guidelines proved difficult. A first realistic goal in such situations is to limit spread of the disease to other areas, specifically to high-risk areas such as intensive care and haemato-oncology units. Optimal management includes a rapid diagnosis of norovirus, written recommendations for management of affected patients and HCWs, and cleaning of surfaces with an effective disinfectant. These recommendations should be available in written form well before such an outbreak is in progress. Such preparations may limit the extent of the outbreak, but norovirus infection in a hospital will probably spread despite infection control interventions. PMID- 14529639 TI - Efficacy of some neutralizers in suspension tests determining the activity of disinfectants. AB - The ability of six mixtures to neutralize glutaraldehyde, o-phthalaldehyde and peracetic acid was tested using four reference strains: Pseudomonas aeruginosa CIP A22, Escherichia coli CIP 54127, Staphylococcus aureus CIP 53154, and Enterococcus faecium CIP 5855. Glutaraldehyde was the hardest to neutralize, and peracetic acid the easiest. The most effective mixture was Tween 80 with sodium bisulphate, sodium thioglycolate, lecithin and cysteine, and the least effective was Tween 80, lecithin and histidine. The efficacy of the neutralizers may indicate a propensity loss of activity from interfering substances when disinfectants are used in practice. PMID- 14529640 TI - Triclosan resistance in methicillin-resistant Staphylococcus aureus (MRSA). AB - Two triclosan selected mutants showed four-fold and 16-fold increases in their minimum inhibitory concentrations (MICs) of triclosan (1 mg/L and 4 mg/L) compared with their parent strains. Four clinical isolates of MRSA were detected with the same triclosan susceptibility as the mutants. One mutant had a predicted change in the gene product on FabI (Thr 147-->His), whilst only one clinical isolate had predicted FabI amino-acid changes (Ala 198-->Gly, and Leu 208-->Phe). The lack of fabI mutations in one mutant and three of the clinical isolates showing reduced triclosan susceptibility suggest that genetic loci other than fabI may be involved in triclosan resistance. PMID- 14529643 TI - A simple system for intraoperative antiseptic wound ventilation. PMID- 14529644 TI - Testing for carriers of Creutzfeldt-Jakob disease. PMID- 14529641 TI - Methicillin-resistant Staphylococcus aureus: prevalence, incidence and risk factors associated with colonization in Sri Lanka. AB - We studied the prevalence of and risk factors for nasal colonization with methicillin-resistant Staphylococcus aureus (MRSA) on admission and the incidence and risk factors for nosocomial acquisition of MRSA in a 60 bed, male surgical unit in the National Hospital of Sri Lanka (NHSL). Nasal swab cultures were obtained from 271 patients admitted for routine surgery within 36 h of admission and repeated every other day until discharge. Clinical and epidemiological risk factors for colonization were compared between patients with MRSA at the time of admission or colonized after admission, and patients who were not colonized. MRSA was isolated from 35 patients (12.9%) with 20 (7.4%) colonized on admission and 15 (6%) acquiring MRSA after admission. Hospitalization within the previous year, antibiotic use within the previous two months, and transfer from another ward within the NHSL were associated with colonization with MRSA on admission. Risk factors for nosocomial acquisition of MRSA were the prophylactic and empiric use of antibiotics. The duration of antibiotic use and the duration of stay in hospital were significantly longer in patients who acquired MRSA. MRSA surveillance and control programmes in this unit would be more cost-effective if targeted at patients with these risk factors. PMID- 14529645 TI - B4 Hand Hygienic Advanced Skin Protection(TM)-a handy adjunct in the prevention of cross-infection? PMID- 14529646 TI - Antibiotic resistance in the community. PMID- 14529647 TI - Last Glacial Maximum ages for robust humans at Kow Swamp, southern Australia. AB - The Kow Swamp people are a fossil population of robust modern humans. We report optically stimulated luminescence (OSL) ages on sediments from Kow Swamp that are at odds with radiocarbon ages obtained previously for the site. The calibrated 14C ages place the Kow Swamp people in the period 15-9 ka. Our single aliquot OSL ages suggest that they lived around the time of the Last Glacial Maximum (LGM) between 22 and 19 ka. An LGM age for the Kow Swamp people is supported by palaeoenvironmental reconstruction. The shoreline silt, in which most of them were interred, was deposited by high lake levels between 26 and 19 ka. Few robust people were left after 19 ka when a sand lunette formed. Climate change may explain the demise of this unusual genetic population. PMID- 14529649 TI - Explaining the end of the hominoid experiment in Europe. AB - The Vallesian Crisis involved the extinction of most of the hominoids that settled successfully in Europe during the middle and early Late Miocene, including Dryopithecus, Ankarapithecus and Graecopithecus. This event has been dated at 9.6 Ma, predating by more than one million years the spread of the C4 grasses and the retreat of forests over large parts of the globe at 7-8 Ma. The finding of macrofloral remains in the Terrassa section (Valles-Penedes Basin) sheds new light on the nature of vegetational change associated with the hominoid extinction. This section presents an abundant late Vallesian vertebrate fauna and has been accurately dated at 9.2 - 9 Ma by paleomagnetism. Therefore, it provides the best indication of the kind of vegetation that occupied the area after the Vallesian Crisis. It is suggested that the extinction of the late Miocene Western European hominoids was not related to the spread of grasses, but to a significant increase of a floral association dominated by deciduous trees. PMID- 14529648 TI - Ancient teeth and modern human origins: an expanded comparison of African Plio Pleistocene and recent world dental samples. AB - Previous research by the first author revealed that, relative to other modern peoples, sub-Saharan Africans exhibit the highest frequencies of ancestral (or plesiomorphic) dental traits and, thus, appear to be least derived dentally from an ancestral hominin state. This determination, in conjunction with various other lines of dental morphological evidence, was interpreted to be supportive of an African origin for modern humans. The present investigation expands upon this work by using: 1) direct observations of fossil hominin teeth, rather than data gleaned from published sources, 2) a single morphological scoring system (the Arizona State University Dental Anthropology System) with consistent trait breakpoints, and 3) data from larger and more varied modern human comparative samples. As before, a multivariate distance statistic, the mean measure of divergence, was used to assess diachronic phenetic affinities among the Plio Pleistocene hominins and modern humans. The present study also employed principal components analysis on dental trait frequencies across samples. Both methods yielded similar results, which support the previous findings; that is, of all modern human samples, sub-Saharan Africans again exhibit the closest phenetic similarity to various African Plio-Pleistocene hominins-through their shared prevalence of morphologically complex crown and root traits. The fact that sub Saharan Africans express these apparently plesiomorphic characters, along with additional information on their affinity to other modern populations, evident intra-population heterogeneity, and a world-wide dental cline emanating from the sub-continent, provides further evidence that is consistent with an African origin model. PMID- 14529650 TI - On the age of Border Cave 5 human mandible. AB - An enamel fragment from the Border Cave 5 specimen was analysed with non destructive ESR combined with laser ablation ICP-MS for uranium profiling. We obtained an age of 74+/-5 ka which fits exactly into the chronological framework that has been previously established for Border Cave by a variety of dating techniques. The result lays at rest the view that BC5 could be of Iron Age, as was implied by (Journal of Human Evolution, 31 (1996) 499) based on nitrogen contents and infra-red splitting factors. PMID- 14529651 TI - 2.6-Million-year-old stone tools and associated bones from OGS-6 and OGS-7, Gona, Afar, Ethiopia. PMID- 14529653 TI - The Early to Middle Stone Age transition and the emergence of modern human behaviour at site 8-B-11, Sai Island, Sudan. PMID- 14529652 TI - Definitive evidence for tail loss in Nacholapithecus, an East African Miocene hominoid. PMID- 14529655 TI - Etiology of epithelial ovarian cancer: a cellular mechanism for the role of gonadotropins. PMID- 14529656 TI - Clinicopathological analysis of c-kit expression in carcinosarcomas and leiomyosarcomas of the uterine corpus. AB - OBJECTIVE: The purpose of this study was to evaluate the expression of the protooncogene, c-kit, in carcinosarcomas and leiomyosarcomas of the uterine corpus and determine the associations between c-kit expression and clinicopathologic factors, including clinical outcome. METHODS AND MATERIALS: Using a polyclonal anti-KIT-antibody, immunohistochemical staining was performed on formalin-fixed paraffin-embedded tissue blocks from 21 carcinosarcomas, 17 leiomyosarcomas, and 1 endometrial stromal sarcoma. KIT-positive tumors were defined as those tumors demonstrating immunopositivity in > or =30% of tumor cells examined. KIT-negative lesions demonstrated immunopositivity in <30% of tumor cells. Two authors independently scored the slides as positive or negative. Staining was repeated on all specimens and independently scored, and in the occasion of a mismatch, a third staining was performed. The carcinosarcomas were catalogued as to whether the sarcomatous and/or carcinomatous elements expressed c-kit. Clinical data were abstracted for those patients with uterine carcinosarcomas. The associations between clinicopathologic characteristics and c kit expression were compared using univariate and multivariate analyses. Kaplan Meier curves based on c-kit expression were plotted for progression-free and overall survival and compared using the log-rank test. RESULTS: Nine of 21 (43%) carcinosarcomas demonstrated immunopositivity for the KIT receptor, although staining was relatively weak. In contrast, only 1/17 (6%) leiomyosarcomas demonstrated KIT immunopositivity (P = 0.029). The solitary endometrial stromal sarcoma evaluated did not demonstrate significant KIT positivity. The majority of KIT-positive carcinosarcomas (6/9 (67%)) demonstrated KIT presence in the sarcomatous portion as compared to the carcinomatous portion (4/9 (44%)). No clinical factor had a statistically significant association with c-kit expression. The lack of c-kit expression was the only factor that was significantly associated with disease recurrence in univariate and multivariate analyses (P < 0.05), although there appeared to be a trend toward a low stage associated with kit positivity. The median progression-free interval for the KIT negative cohort was 8 months, while it had not been reached for the KIT-positive cohort after median follow-up of 15 months (P = 0.0462). CONCLUSIONS: A significant proportion of carcinosarcomas of the uterine corpus display immunoreactivity for c-kit. Patients with KIT-positive carcinosarcomas may have an improved progression-free survival compared to KIT-negative tumors; however, further data are needed to determine whether this finding is confounded by stage. PMID- 14529657 TI - Uterine sarcomas express KIT protein but lack mutation(s) in exon 11 or 17 of c KIT. AB - OBJECTIVE: Several tumors express the protein product of the protooncogene c-KIT. Some of these respond to imatinib mesylate, a tyrosine kinase inhibitor. The tumors that respond frequently have mutation(s) in exon 11 of c-KIT that encodes for the regulatory juxtamembrane helix. Some tumors that express KIT protein have mutation(s) in exon 17 of c-KIT; however, these do not respond to imatinib mesylate. This investigation was performed to determine the expression of KIT protein and mutational status of exons 11 and 17 of c-KIT in uterine sarcomas. METHODS: Twenty-five uterine sarcomas treated from 1990 to 2002 were evaluated. These included 14 malignant mullerian mixed tumors (MMMT), 7 leiomyosarcomas (LMS), 2 endometrial stromal sarcomas (ESS), and 2 high-grade heterologous sarcomas (HGHS). Formalin-fixed, paraffin-embedded tissue sections were immunostained with anti-KIT antibody (Santa Cruz Biotechnology, Santa Cruz, CA) with a semiquantitative assessment. Normal myometrium when present in the section was used as an internal negative control. Areas of tumor were microdissected followed by DNA extraction, polymerase chain reaction (PCR) amplification of exons 11 and 17, single-strand conformational polymorphism (SSCP), and DNA sequencing to detect the presence of mutation(s). RESULTS: All 25 tumors expressed KIT protein at varying levels as assessed by immunohistochemistry. The staining was diffuse and of moderate to strong intensity in 22 tumors. In three tumors (one of each type except MMMT) the staining intensity was weak. In MMMT the epithelial and sarcomatous foci stained similarly. No mutation(s) in exons 11 or 17 of c-KIT were identified in 24/25 tumors. One LMS had deletion of both exons 11 and 17. CONCLUSIONS: Although uterine sarcomas express KIT protein, they lack KIT-activating mutation(s) in exon 11 or 17 of c-KIT. Therefore, these tumors are unlikely to respond to imatinib mesylate. PMID- 14529658 TI - Ovarian cancer and high-risk women-implications for prevention, screening, and early detection. AB - OBJECTIVE: The aim of this study was to understand the strengths and limitations of current prevention, detection, and screening methods for ovarian cancer and to identify research areas to improve prevention, screening, and detection of the disease for all women as well as for women carrying a mutation in the BRCA1/2 genes. METHODS: We convened an ovarian cancer symposium at the University of Pittsburgh in May 2002. Nineteen leading scientists representing disciplines such as epidemiology, molecular biology, pathology, genetics, bioinformatics, and psychology presented the latest data on ovarian cancer prevention, screening, and early detection. RESULTS: Ovarian cancer is the most common cause of death from a gynecologic malignancy in the United States. Because survival depends on stage of diagnosis, early detection is critical in improving clinical outcome. However, existing screening techniques (CA125, transvaginal ultrasound) have not been shown to reduce morbidity or mortality. Moreover, with the exception of oral contraceptives, there are no available chemopreventive agents. Bilateral salpingo oophorectomy also has been shown to reduce incidence, but this procedure has several drawbacks in terms of a woman's reproductive, cardiovascular, skeletal, and mental health. CONCLUSIONS: Better methods to prevent, detect, and screen for ovarian cancer in all women, but particularly in high-risk women carrying mutations in BRCA1/2, are urgently needed. This article reviews the current state of knowledge in the etiology, prevention, and early detection of ovarian cancer and suggests several areas for future clinical, epidemiologic, and laboratory based research. PMID- 14529659 TI - Gemcitabine combined with cisplatin as first-line treatment in patients with epithelial ovarian cancer: a phase II study. AB - OBJECTIVE: This phase II study was performed to evaluate the activity and toxicity of gemcitabine plus cisplatin as first-line treatment of advanced epithelial ovarian cancer. METHODS: Chemonaive patients with histologically or cytologically confirmed FIGO stage III or IV epithelial ovarian carcinoma were enrolled. Patients received cisplatin 75 mg/m(2) on Day 1 and gemcitabine 1250 mg/m(2) on Days 1 (after cisplatin) and 8 of a 21-day cycle. RESULTS: Of the 42 female patients (median age 60 years) enrolled, 81% had a Zubrod performance status of 0 or 1. Among the 37 response-evaluable patients, there were 5 (13.5%) pathological complete responses (CRs), 16 (43.2%) pathological partial responses (PRs), and 3 (8.1%) clinical PRs, for an overall response rate of 64.9% (95% CI: 47.4-79.8%) and a pathological response rate of 56.8%. Per an intent-to-treat analysis, the overall response rate was 57.1% (95% CI: 41.0-72.3%). After a median follow-up time of 15.8 months, the median survival was 24.0 months and median progression-free survival was 13.4 months. Grade 3/4 neutropenia and thrombocytopenia occurred in 69.0 and 33.3% of patients, respectively, with no febrile neutropenia or hemorrhage. Grade 3/4 nausea and vomiting occurred in 35.7% and grade 3 alopecia in 21.4% of the patients. One patient died due to a toxicity-related death (dyspnea). CONCLUSIONS: Gemcitabine plus cisplatin is active and feasible as first-line treatment of advanced epithelial ovarian cancer. Further clinical trials adding gemcitabine to first-line treatment seem warranted. PMID- 14529660 TI - Conventional 3D conformal versus intensity-modulated radiotherapy for the adjuvant treatment of gynecologic malignancies: a comparative dosimetric study of dose-volume histograms. AB - OBJECTIVE: The goals of this study were to evaluate the feasibility of pelvic intensity-modulated radiotherapy (IMRT) in the adjuvant treatment of gynecologic malignancies and to compare the dose-volume histograms (DVHs) and determine the potential impact on acute and long-term toxicity based on the dose to target and nontarget tissues for both planning techniques. METHODS: Ten consecutive patients referred for adjuvant radiotherapy for gynecologic malignancies at the University of Pittsburgh School of Medicine and Magee-Womens Hospital were selected for CT based treatment planning using the ADAC 3D version 4.2g and the NOMOS Corvus IMRT version 4.0. Normal tissues and critical structures were contoured on axial CT slices by both systems in conjunction with a gynecologic radiologist. These regions included internal, external, and common iliac nodal groups, rectum, upper 4 cm of vagina, bladder, and small bowel. Conventional treatment planning included 3D four-field box using 18-MV photons designed to treat a volume from the L(5)/S(1) border superiorly to the bottom of the ischial tuberosity on the AP/PA field and shaped blocks on the lateral fields to minimize the dose to the rectum and small bowel. A seven-field technique using 6-MV photons was used for IMRT. Restraints on small bowel for IMRT were set at 23.0 Gy +/- 5% and 35.0 Gy+/ 5% for the rectum and 37.5 Gy +/- 5% for the bladder while simultaneously delivering full dose (45.0 Gy) to the intrapelvic nodal groups in 1.8-Gy daily fractions. The dose-volume histograms where then compared for both treatment delivery systems. RESULTS: The volume of each organ of interest (small bowel, bladder, and rectum) receiving doses in excess of 30 Gy was compared in the 3D and IMRT treatment plans. The mean volume of small bowel receiving doses in excess of 30 Gy was reduced by 52% with IMRT compared with 3D. A similar advantage was noted for the rectum (66% reduction) and the bladder (36% reduction). The nodal regions at risk and the upper vagina all received the prescribed dose of 45.0 Gy. CONCLUSIONS: Intensity-modulated radiotherapy appears to offer several advantages over conventional 3D radiotherapy (3D CRT) planning for adjuvant radiotherapy for gynecologic malignancies. These include a significant reduction in treatment volume for bladder, rectum, and small bowel. It is anticipated that this reduction in volume of normal tissue irradiated would translate into overall reduction in acute and potentially late treatment-related toxicity. Prospective trials are necessary to better evaluate the advantages in a larger group of patients. PMID- 14529661 TI - Preoperative differentiation of malignant from benign ovarian tumors: the efficacy of morphology indexing and Doppler flow sonography. AB - OBJECTIVE: The goal of this study was to determine the efficacy of morphology indexing and Doppler flow sonography as methods to predict risk of malignancy in sonographically confirmed ovarian tumors. METHODS: Risk of malignancy was assessed preoperatively in 442 ovarian tumors using a new morphology index (MI) based on tumor volume and wall structure. Each tumor was assigned a score of 0 to 10 based on increasing volume and morphologic complexity. Doppler flow studies were performed on 371 of these tumors. Following morphologic evaluation, all ovarian tumors were removed surgically. RESULTS: Of 315 tumors with a MI < 5 there was only 1 malignancy (a stage IA granulosa cell tumor <2 cm in diameter) whereas there were 52 malignancies in 127 tumors with a MI > or = 5. Stage of disease was as follows: stage I, 33; stage II, 6; stage III, 14. Risk of malignancy was related directly to MI score, varying from 0.3% in tumors with a MI < 5 to 84% in tumors with a MI > or = 8. A MI value of > or = 5 as indicative of malignancy was associated with the following statistical parameters: sensitivity 0.981, specificity 0.808, PPV 0.409, NPV 0.997. A pulsatility index (PI) < 1.0 as indicative of malignancy was associated with: sensitivity 0.528, specificity 0.776, PPV 0.288, NPV 0.906. A resistive index (RI) < 0.4 as indicative of malignancy was associated with: sensitivity 0.222, specificity 0.867, PPV 0.222, and NPV 0.867. The addition of Doppler flow indices to MI did not improve the accuracy of predicting malignancy. Likewise, the absence or presence of ovarian tumor blood flow was not reliable as a means to differentiate benign from malignant ovarian tumors. CONCLUSIONS: Morphology indexing is an accurate and inexpensive method of differentiating benign from malignant ovarian tumors, and can be a valuable adjunct in treatment planning. The addition of Doppler flow studies did not improve diagnostic accuracy of MI. PMID- 14529662 TI - Co-overexpression of p53 and bcl-2 proteins in HPV-induced squamous cell carcinoma of the uterine cervix. AB - OBJECTIVE: The aim of this study was to analyze aberrant expression of both apoptotic protein p53 and antiapoptotic protein bcl-2 in squamous cell carcinoma (SCC) of the uterine cervix with HPV infection and its significance as a marker for progression of cervical lesions. METHODS: One hundred and five cervical lesions and 20 normal (age matched) cervical epithelium from patients with complaints other than cervical lesions were investigated immunocytochemically for aberrant expression of p53 and bcl-2 using the streptavidin-biotin-peroxidase method with respective monoclonal antibodies. HPV status was also anlayzed using type-specific primers for HPV 16/18 and HPV 6/11 by polymerase chain reaction (PCR). The statistical correlation analysis was carried out using Spearman's correlation test and univariate analysis by the SPSS system. RESULTS: An abnormal nuclear expression of tumor-suppressor protein p53 and cytoplasmic expression of bcl-2 were observed using immunocytochemistry in biopsies of cervical lesions but not in normal subjects. The intensity of immunoreactivity for both p53 and bcl-2 proteins varied between different histopathological grades of cervical lesions and the correlation analysis showed a highly significant positive correlation for their expression level with different stages from mild dysplasia to invasive cancer with r = 0.88842; P = 0.00001 and r = 0.86929; P = 0.00001, respectively. A highly significant positive correlation was also observed between the expression of both p53 and bcl-2 proteins and HPV infection. The current study indicates a very good significant direct correlation (r = 0.83925; P = 0.00001) between p53 expression and bcl-2 expression in the study population, suggesting the co-overexpression of these proteins in HPV-associated cervical cancer. CONCLUSIONS: From the observations it is suggested that the immunodetection of both p53 and bcl-2 proteins in squamous cell carcinoma of the uterine cervix can be used as an independent diagnostic marker for cervical cancer associated with HPV infection. The highly significant association of these proteins with HPV infection suggests that the high-risk HPV infection may be responsible for the co overexpression of p53 and bcl-2 in cervical cancer even though both of them are antagonistic in their function. This study thus helps to understand the molecular mechanism underlying cervical carcinogenesis and which in turn may improve the therapeutic approach. PMID- 14529663 TI - Computed tomography and magnetic resonance imaging in staging of uterine cervical carcinoma: a systematic review. AB - OBJECTIVE: The goal of this article is to systematically review the available evidence on the diagnostic performance of computed tomography (CT) and magnetic resonance imaging (MRI) in staging of cervical carcinoma. METHODS: A comprehensive computer literature search was performed in MEDLINE and EMBASE databases from January 1985 to May 2002. Two reviewers independently scored methodological quality of included studies and extracted relevant data for data analysis. A bivariate random effect approach was used to summarize estimates of sensitivity and specificity values. Covariates were added to this model to study the influence of sample size, publication year, methodological criteria, and MRI techniques on summary estimates. RESULTS: Fifty-seven articles were included. In 49 articles one imaging modality was evaluated (MRI, 38; CT, 11), and in 8 articles, both. Inclusion criteria were: minimum of 10 patients included, histopathology as reference standard, sufficient data presented to construct 2(x) 2 tables. The exclusion criterion was: data reported elsewhere in more detail. Sensitivity estimates for parametrial invasion were 74% (95% C: 68-79%) for MRI and 55% (95% CI: 44-66%) for CT, and for lymph node involvement, 60% (95% CI 52% 68%) and 43% (95% CI: 37-57%), respectively. MRI and CT had comparable specificities for parametrial invasion and lymph node involvement. For bladder invasion and rectum invasion the sensitivities for MRI were respectively 75% (95% CI: 66-83%) and 71% (95% CI: 53-83%), higher compared with CT. The specificity in evaluating bladder invasion for MRI was significantly higher compared with CT: 91% (95% CI: 83-95%) for MRI and 73% (95% CI: 52-87%) for CT. The specificities for rectum invasion were comparable. Differences in patient sample size, publication year, methodological criteria, and MRI techniques had no effect on the summary estimates. CONCLUSIONS: For overall staging of cervical carcinoma, MRI is more accurate than CT. PMID- 14529664 TI - HPV testing can reduce the number of follow-up visits in women treated for cervical intraepithelial neoplasia grade 3. AB - OBJECTIVE: We evaluated high-risk human papillomavirus (HPV) testing by Hybrid Capture II (HC II) in addition to cytology to predict recurrent/residual cervical intraepithelial neoplasia (CIN) 2/3 and cervical cancer in women treated for CIN 3. METHODS: Follow-up study of 108 women with histologically confirmed CIN 3. RESULTS: Pretreatment, in 96% (104/108) of the smears high-risk HPV DNA was present. Posttreatment, 71% (77/108) of the women had normal cytology and negative HC II test and none developed recurrent/residual disease during a median follow-up of 28.8 months with a range of 2.4-64.8 months. One of the 12% (13/108) of women with normal cytology and positive HC II test was diagnosed with cervical adenocarcinoma. One of the 7% (8/108) of women with abnormal cytology (borderline dyskaryosis or worse) and negative HC II test was diagnosed with CIN 2. Three of the 9% (10/108) of women with abnormal cytology and a positive HC II test were diagnosed with CIN 2/3. These results show an increased risk for recurrent/residual CIN 2/3 and cervical carcinoma when at least one posttreatment test is positive. The highest relative risk (72.9, 95% CI 25-210) was present in women with both tests positive. CONCLUSIONS: HPV testing with Hybrid Capture II in conjunction with cytology can be used as a tool to select women with an increased risk for recurrent/residual CIN 2/3 and cervical cancer. The standard policy in The Netherlands is cytology at 6, 12, and 24 months posttreatment. However, for women with both normal cytology and negative HC II test at 6 months the chance to develop recurrent/residual CIN 2/3 and cervical carcinoma is so low that retesting at 12 months can be omitted. PMID- 14529665 TI - Gynecologic screening in hereditary nonpolyposis colorectal cancer. AB - OBJECTIVE: In hereditary nonpolyposis colorectal cancer (HNPCC), women with a mismatch repair (MMR) gene mutation have a cumulative lifetime risk of 25-50% for endometrial cancer and 8-12% for ovarian cancer. Therefore, female members of HNPCC families are offered an annual gynecologic and transvaginal ultrasound (TVU) examination and serum level CA 125 analysis. The aim of the present study was to evaluate our 10-year experience with this screening program. METHODS: Women who are MMR gene mutation carriers or who fulfil the Amsterdam criteria were identified from our HNPCC database. Information concerning the screening program was retrospectively collected from patient files. RESULTS: Forty-one women, 35 premenopausal and 6 postmenopausal, were enrolled in the program with a median follow-up of 5 years (range 5 months-11 years). In 197 patient years at risk, 17 of 179 TVUs gave reason for endometrial sampling. Three premalignant lesions, with complex atypical hyperplasia, were discovered. One interval endometrial cancer was detected as a result of clinical symptoms. No abnormal CA 125 levels were measured and no ovarian cancers were detected. CONCLUSIONS: These results demonstrate that gynecologic screening allows the detection of premalignant lesions of the endometrium but also illustrate that recognition and reporting of clinical symptoms by the women themselves is of utmost importance. PMID- 14529666 TI - Oncolytic viral therapy for human ovarian cancer using a novel replication competent herpes simplex virus type I mutant in a mouse model. AB - OBJECTIVE: Attenuated mutant strains of herpes simplex virus (HSV) have been effectively used for treatment of malignant brain tumors. As HSV-1 can infect and lyse a variety of cell types, other malignancies may also benefit from such treatment. We sought to test the feasibility of HSV-1 mutant-mediated gene therapy treatment of ovarian cancer. METHODS: We prepared two attenuated mutant HSV-1 strains. An HSV-1 mutant, hrR3, has replaced the gene encoding ribonucleotide reductase (RR) with the lacZ reporter gene. We also developed a new replication-competent HSV-1 mutant, HR522; this virus, expressing the lacZ reporter gene, induces syncytium formation in infected cells. We compared the efficacy of HR522 with, paclitaxel (Taxol) and hrR3 in the treatment of nude mice harboring human ovarian cancer cells. We also examined the effect of the prodrug ganciclovir (GCV) on the treatment mediated by these HSVs. Survival was evaluated by Kaplan-Meier method and log-rank test. RESULTS: The survival of mice treated with a high-titer hrR3 (5 x 10(7) plaque-forming units [PFU]) was significantly prolonged as compared with the group given paclitaxel (P < 0.0001, log-rank test). Although the survival of mice treated with high-titer HR522 (5 x 10(7) PFU) was not significantly prolonged compared with paclitaxel-treated group (P = 0.212, log-rank test), GCV markedly enhanced the efficacy of HR522 administration (P < 0.005, vs paclitaxel, log-rank test). The lacZ gene product, visualized using 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) histochemistry, was detected in HR522-treated tumors in areas also exhibiting apoptotic changes. CONCLUSIONS: These findings indicate that the combination of HR522 and GCV possesses significant therapeutic potential for treatment of ovarian cancer. Such viral therapy offers a novel approach to reductions in the dissemination of ovarian cancer. PMID- 14529667 TI - Resistance to Fas-mediated cell death in BeWo and NJG choriocarcinoma cell lines: implications in immune privilege. AB - OBJECTIVE: An immune privileged site occurs when the allogenic tissue grafts have the propensity for prolonged survival in the host tissue. In this context, the survival and proliferation of malignant trophoblasts in the gravid uterus are currently unclear. In a previous study, we documented that Fas and FasL are coexpressed in choriocarcinoma [Gynecol. Oncol. (2003)]. This study was conducted to examine the role of the Fas/FasL pathway in immune privilege of BeWo and NJG choriocarcinoma cells in culture. METHODS: The ability of anti-Fas mAb (CH-11) to sensitize choriocarcinoma cell lines to Fas-mediated cytotoxicity was assessed by MTT assays. Coculture experiments with Fas-sensitive Jurkat cells were used to demonstrate functional FasL from choriocarcinoma. RT-PCR was used to assess the expression of cFLIP. RESULTS: The mean cell viability of BeWo and NJG cells declined to about 58 and 63% compared to controls after 72 h of culture in the presence of anti-Fas mAb (CH-11) while the Fas-sensitive Jurkat cells showed viability of only 10%. This resistance to Fas-mediated apoptosis in choriocarcinoma cells is reversed in the presence of cycloheximide (0.5 micro g/ml) which further decreased the viability to 36 and 32%, respectively, at a dose of 300 ng/ml (P < 0.05). The observed resistance to Fas-mediated apoptosis therefore could be attributed to the short-lived endogenous inhibitor, cFLIP as demonstrated by the RT-PCR technique. In coculture experiments, FasL from choriocarcinoma cells induced apoptosis in the Fas-sensitive Jurkat cells, thereby indicating the capacity to evade immune attack. CONCLUSIONS: Decreased sensitivity to Fas-mediated apoptosis and counterattacking the lymphocytes may impart immune privilege in these malignant trophoblasts for prolonged survival in the host. PMID- 14529668 TI - Co-expression of Fas (APO-1, CD95)/Fas ligand by BeWo and NJG choriocarcinoma cell lines. AB - OBJECTIVE: Fas (CD95) is a transmembrane protein of the tumor necrosis factor receptor superfamily that induces apoptosis in susceptible cells on crosslinking by its ligand (FasL). The Fas loss of function and concurrent expression of its ligand (FasL) have been associated with malignant phenotype. In this study, we sought to investigate the hitherto undescribed expression of Fas and FasL on the immortalized human choriocarcinoma cell lines BeWo and NJG. METHODS: Receptor and ligand expression was demonstrated using specific antibodies and multiple techniques including immunocytochemistry, confocal immunofluorescence microscopy, flow cytometry, immunoblots, and reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Data from this study indicate that human choriocarcinoma cell subtypes co-express both Fas and FasL. A specific cytoplasmic and membranous pattern of immunoreactivity was noted that was further confirmed at mRNA transcripts by RT-PCR. In addition, we provide evidence using flow cytometry that the Fas receptors are downregulated. The mean fluorescence intensities for NJG and BeWo were 1.47 +/- 0.5 and 1.59 +/- 0.4, while that for Fas-positive Jurkat cells was 25.6 +/- 3.1. CONCLUSIONS: To our knowledge, this is the first report on the identification and constitutive co-expression of Fas and FasL in BeWo and NJG choriocarcinoma cells. Choriocarcinoma cells evade immune attack by downregulating the Fas receptor and by killing lymphocytes through expression of FasL. Taken together, our investigations suggest that the Fas/FasL system may represent a mechanism by which malignant trophoblasts become resistant to apoptosis, escape immune surveillance, and metastasize. PMID- 14529669 TI - High preoperative plasma concentration of tissue plasminogen activator (tPA) is an independent marker for shorter overall survival in patients with ovarian cancer. AB - OBJECTIVE: The objective was to evaluate the preoperative blood concentration of tissue plasminogen activator (tPA) as a discriminator between malignant and benign ovarian tumors, and as a potential marker of postoperative prognosis in patients with ovarian cancer. METHODS AND MATERIAL: The concentration of tPA was assayed with ELISA (Imulyse Biopool) in preoperative plasma samples obtained from 111 patients with adnexal lesions. Tumors were classified as benign (n = 25), borderline malignant (n = 11), well-differentiated (G1, n = 22), moderately differentiated (G2, n = 11), and poorly differentiated malignant (G3, n = 42). The median follow-up time of patients with malignant tumors was 5.6 years (range 2.1-13.2 years) and 37 patients died during the follow-up period. RESULTS: Patients with moderately and poorly differentiated tumors had higher levels of plasma tPA compared to those with well-differentiated tumors (P = 0.004 and P = 0.005). No significant differences in the plasma tPA levels were observed between patients with benign, borderline, and well-differentiated tumors. The tPA levels were not different between stages nor within stage Ia-c. In a multivariate Cox proportional hazards model including stage, grade, age, and plasma tPA dichotomized at the median (> or =9 vs <9 ng/mL), high levels of tPA were significantly associated with shorter survival: HR = 4.4 (95% CI 2.0-9.8, P = 0.0003). In the univariate analyze high levels of tPA showed HR = 4.5 (95% CI 2.1 9.6, P = 0.0003). CONCLUSIONS: High concentration of plasma tPA was an independent marker for poor prognosis in patients with ovarian cancer in our study. Plasma tPA did, however, not discriminate between benign and malignant adnexal lesions. PMID- 14529670 TI - Carboplatin plus paclitaxel combination chemotherapy: impact of sequence of drug administration on treatment-induced neutropenia. AB - OBJECTIVE: While the importance of the sequence of administration of cisplatin and paclitaxel on the degree of observed neutropenia has been documented, there is limited information available in the oncology literature to determine whether there exists sequence-dependent toxicity for the combination of carboplatin plus paclitaxel. METHODS: Patients with advanced gynecologic malignancies were randomized to receive either carboplatin (AUC 6), followed by paclitaxel (175 mg/m(2) over 3 h) (C-P), or the same doses of the agents delivered in the opposite sequence (P-C). The primary endpoint was the degree of neutropenia experienced during the initial treatment course. RESULTS: A total of 40 patients (median age: 63) entered this trial, of whom 27 had complete pretreatment and nadir counts available for course 1 and 24 for both course 1 and course 2. By random chance, patients initially receiving P-C began therapy with a higher baseline ANC than those treated with C-P. During course 1, the P-C population was noted to have a greater reduction, from baseline, in ANC (P = 0.02), but no difference in absolute nadir counts (ignoring the baseline value) (P = 0.64). There was no difference between P-C, followed by C-P, versus C-P, followed by P-C in the severity of neutropenia experienced during course 2 (P = 0.38). CONCLUSIONS: The sequence of carboplatin/paclitaxel administration does not exert a significant influence on the level of observed neutropenia. This finding leads to the suggestion that the sequence of drug delivery can be modified, as necessary, to satisfy unique requirements of individual patients and to establish the optimal drug delivery strategy of an innovative investigational treatment regimen. PMID- 14529671 TI - Platinum retreatment of platinum-resistant ovarian cancer after nonplatinum therapy. AB - OBJECTIVE: The objective was to determine the response rate to platinum retreatment of "platinum-resistant" ovarian cancer after intervening nonplatinum therapy. METHODS: We retrospectively identified 30 patients with platinum resistant ovarian cancer who received nonplatinum chemotherapy for recurrent epithelial ovarian cancer prior to additional platinum therapy. All patients were treated between July 1, 1997, and June 30, 2001. Platinum resistance was defined as less than a partial response to platinum therapy or progression within 6 months of the last platinum therapy. RESULTS: Overall, 7 of 30 patients experienced an objective response to platinum therapy (partial response, 23%; complete response, 0%) based on CT scan (2/21) and/or CA-125 (5/9) criteria. The median time to progression for the group was 17 weeks (range, 4-59 weeks). Several predictive factors were identified. The interval since the last platinum treatment did not appear to be predictive in this group. Only 1 of 16 patients who did not have an objective response to the most recent platinum-based therapy responded to platinum rechallenge. Similarly, no patient who received more than three intervening nonplatinum treatments responded to additional platinum therapy (0/10). CONCLUSIONS: Our small retrospective series suggest that the platinum resistant category is heterogenous and includes patients who may respond to retreatment with platinum-based agents. This group includes the patients with prior platinum responses and early progression. However, patients without an objective response to the last prior platinum therapy or more than three intervening treatments are unlikely to respond to subsequent platinum therapy. PMID- 14529672 TI - Laser CO2 vaporization for high-grade cervical intraepithelial neoplasia: a long term follow-up series. AB - OBJECTIVE: The goal of this study was to evaluate the effectiveness of laser CO(2) vaporization for conservative treatment of ectocervical high-grade cervical intraepithelial neoplasia (CIN) particularly by the evaluation of the reappearance risk of disease in long-term follow-up. METHODS: One hundred fifty nine patients were submitted to CO(2) laser vaporization for high-grade CIN and followed up for a minimum of 5 years. Selection of cases, depth of ablation, complications, and cure rate (percentage of treated patients in whom there was no recurrent/persistent high-grade CIN at the 5-year follow-up examination) were retrospectively evaluated. RESULTS: Selected cases for colposcopy were submitted to a 6-mm mean depth of vaporization without intra- or postoperative complications. The cure rate for a single treatment was 97.5% and a satisfactory colposcopic follow-up was possible in 99.4% of treated patients. No case of invasive carcinoma occurred after a mean follow-up of 7.1 years. Four cases (2.5%) were high-grade CIN persistence observed after a mean time of 3.75 months, suggesting incomplete destruction of the deepest part of the lesion involving the glandular crypt base. CONCLUSIONS: Long-term follow-up proves that laser CO(2) vaporization still has a place in the treatment of CIN. In selected cases it represents a safe alternative for conization in the treatment of high-grade CIN, but colposcopic expertise is essential for adequate preoperative selection of cases. PMID- 14529673 TI - Liquid-based cytology for the postirradiation surveillance of women with gynecologic malignancies. AB - OBJECTIVE: To determine the performance of liquid-based cytology using ThinPrep in the postirradiation surveillance of women with gynecologic malignancies. METHODS: Patients with endometrial and cervical cancer treated with primary or adjuvant radiotherapy between 2000 and 2002 were identified. Details regarding tumor characteristics, treatment, and cytologic and histologic results were abstracted. Binomial variables were compared with the chi-square test. The performance characteristics of liquid-based cytology were evaluated. RESULTS: A total of 302 liquid-based cytologic samples from 121 women were evaluated. Overall, 294 (97.4%) of the specimens were adequate for interpretation and 132 (44.9%) were within normal limits. Benign cellular changes, including benign radiation changes, were identified in 141 (47.6%). Atypical squamous cells (ASCUS) were found in 15 (5.1%), recurrent dysplasia in 4 (1.3%), and recurrent carcinoma in 2 (0.7%). Follow-up of the 15 ASCUS smears revealed 13 (86.7%) normal smears and 2 cases of squamous intraepithelial lesions. Benign findings were noted in three of the four smears with SIL. The sensitivity for the detection of SIL was 33%, the specificity 99%, and the positive predictive value (PPV) 25%. Of the 4 patients with local recurrences, 2 were detected by cytology. The sensitivity for the detection of recurrent carcinoma was 50%, with a specificity and PPV of 100%. CONCLUSIONS: ThinPrep for the surveillance of women with gynecologic malignancies treated with radiotherapy is associated with a high rate of satisfactory samples and a low rate of equivocal and ASCUS cytology. PMID- 14529674 TI - Laparoscopic paraaortic left-sided transperitoneal infrarenal lymphadenectomy in patients with gynecologic malignancies: technique and results. AB - OBJECTIVE: Left-sided paraaortic infrarenal lymphadenectomy is indicated in patients with gynecologic tumors of high metastasing potential. We evaluated whether left-sided paraaortic inframesenteric lymphadenectomy can be extended up to the left renal vein by laparoscopy. METHODS: Between January 2002 and August 2002, 46 consecutive patients with cervical (n = 26), or endometrial (n = 16), or early ovarian cancer (n = 4) underwent right-sided paraaortic lymphadenectomy up to the level of the right ovarian vein and left-sided inframesenteric paraaortic lymphadenectomy. Lymphadenectomy was extended up to the level of the left renal vein in 20 patients with high risk for lymph node metastasis: following elevation of the duodenum and the pancreas infrarenal lymph nodes in the area limited by the vena cava, left renal vein, left ovarian vein, inframesenteric artery, and aorta were laparoscopically removed under preservation of the inferior mesenteric artery. RESULTS: Patients with infrarenal lymphadenectomy (group 1) and without infrarenal lymphadenectomy (group 2) were comparable in body mass index: the age of patients in group 2 was higher (P = 0.023). Duration of lymphadenectomy was 31.3 min (11-57 min) longer in group 1. There was no intraoperative complication. Number of paraaortic lymph nodes was on average 19.6 (range 5-35) in group 1 compared to a mean of 9 lymph nodes (range 2-19) in group 2 (P = 0,0001). Postoperatively 2 patients (10%) in group 1 developed chylascos. CONCLUSIONS: Left-sided paraaortic infrarenal lymphadenectomy can be performed safely in adequate duration transperitoneally by laparoscopy. Compared to inframesenteric lymphadenectomy the number of removed lymph nodes can be doubled. PMID- 14529675 TI - Follow-up by combined cytology and human papillomavirus testing for patients post cone biopsy: results of a long-term follow-up. AB - OBJECTIVE: The goal of this study was to evaluate the clinical implications of integrating human papillomavirus (HPV) testing into a long-term follow-up and management protocol for women postconization for high-grade cervical intraepithelial neoplasia (CIN2-3). METHODS: Sixty-seven women were followed-up by Pap smears and HPV type and load testing (mean follow-up, 63 months; range, 50 72). Patients with persistent abnormal cytology on two consecutive smears and those with positive HPV test results (whatever their cytologic findings) were referred for colposcopy-directed biopsy. Patients histologically diagnosed with CIN2-3 and those with high-load HPV (whatever their histologic findings) underwent repeat conization or hysterectomy for residual disease. RESULTS: At follow-up, 29 (43.2%) women had positive cytology or positive HPV results and were referred for colposcopy. Eleven (37.9%) had high-grade cervical intraepithelial neoplasia or high-load HPV results and were further treated by reconization/hysterectomy. The respective positive predictive values of high-load HPV and low-grade squamous intraepithelial lesions were 100 and 60% for any CIN and 90 and 15% for CIN2-3. Only five of nine cases with a final diagnosis of CIN2 3 were originally identified by cytology: the other four were detected only by parallel evaluation by HPV testing. High-load HPV results with normal cytology or low-grade lesions harbored an 80% risk for CIN2-3. CONCLUSIONS: Adding HPV load assessment to the follow-up protocol of women postconization due to CIN2-3 lesions could help detect high-grade residual disease among low-grade lesions and normal cytology cases while concomitantly and safely bestowing the advantage of lowering the rates of colposcopic referrals and surgical procedures. PMID- 14529676 TI - Prospective longitudinal study of ultrasound screening for endometrial abnormalities in women with breast cancer receiving tamoxifen. AB - OBJECTIVE: The goal of this work was to study the role of transvaginal ultrasonography (TVUS) together with colorflow Doppler imaging (CFDI) in the detection of significant endometrial abnormalities induced by tamoxifen. METHODS: Over a 6-year period, 304 women on tamoxifen as adjuvant therapy for breast cancer were recruited into the current study. Standard demographic data as well as duration of tamoxifen use were collected. Patients were assessed at study entry and at yearly intervals with TVUS together with CFDI. All patients had an endometrial biopsy at the time of study entry, and repeat endometrial evaluations were done subsequently only if there were abnormal ultrasound findings or the presence of irregular vaginal bleeding. All ultrasonic characteristics and Doppler flow measurements were recorded. Descriptive statistics were used to describe the study group. Logistic regression was used to identify significant treatment- and ultrasound-related factors associated with the presence of significant uterine pathology. RESULTS: One thousand and sixty-one ultrasound assessments were performed on 304 patients over a 6-year period. The mean age was 52.33 (range, 29-79). Seventy-two percent of the patients were postmenopausal at the time of breast cancer diagnosis. The median concentrations of estrogen and progesterone receptor were 75 and 73 fmol/L, respectively. Fifty-eight percent of the patients had received cytotoxic chemotherapy. The mean duration of tamoxifen use was 48.2 months. Thirty-two percent of the ultrasound examinations had associated significant uterine pathology defined as conditions that required further medical or surgical investigation and treatment. However, 80% of the abnormalities represented benign polyps. Six cases of primary endometrial cancer were detected. All cases presented with irregular bleeding. No recurrence of disease was detected at a median follow-up of 48 months. One case of metastatic breast cancer to the uterus was encountered. By setting the endometrial thickness cutoff at more than 9 mm to represent significant abnormality in this patient population, the sensitivity was 63.3%, specificity was 60.4%, positive predictive value was 43.3%, and negative predictive value was 77.5%. To detect endometrial cancer, the endometrial thickness cutoff at 9 mm had a positive predictive value of only 1.4%. Logistic regression analysis showed only endometrial thickness greater than 9 mm (OR 3.99, CI = 1.26-12.65, P = 0.018) and spiral artery pulsatility index measurement (OR 4.18, CI = 1.25-13.92, P = 0.02) to be associated with significant uterine abnormalities. CONCLUSIONS: Routine sequential ultrasound surveillance in asymptomatic women on tamoxifen is not useful because of its low specificity and positive predictive value. A significant portion of screened asymptomatic women would need to undergo needless surgical evaluations of their endometrium if widespread use of ultrasound is implemented in this patient population. PMID- 14529677 TI - Mitogen-activated protein kinases (MAPK) as predictors of clinical outcome in serous ovarian carcinoma in effusions. AB - OBJECTIVE: The objective was to investigate the expression (level) and phosphorylation status (activity) of the extracellular-regulated kinase (ERK), c Jun amino-terminal kinase (JNK), and high-osmolarity glycerol response kinase (p38), their role in the biology of ovarian carcinoma, and their correlation with chemotherapy response. METHODS: Sixty-four fresh-frozen effusions from patients diagnosed with serous ovarian carcinoma were studied using immunoblotting. Results were analyzed for possible association with expression of proliferation and apoptosis markers, patient age, disease stage, tumor grade, histological grade, chemotherapy status, and survival. RESULTS: p38 level correlated with younger age (P = 0.004), while that of JNK correlated with better tumor differentiation (P = 0.009). Higher expression of Pan-JNK (P = 0.018) and higher p-ERK activity (P = 0.014) were seen in postchemotherapy specimens, specifically related to treatment by platinum agents. pan-JNK expression was higher in specimens treated with both platinum agents (P = 0.038) and paclitaxel (P = 0.033). In univariate survival analysis, the level of pan-ERK (P = 0.002), pan JNK (P = 0.045), and pan-p38 (P = 0.016), as well as p-ERK activity (P = 0.014) correlated with better overall survival. In Cox multivariate survival analysis, pan-ERK (P = 0.001), pan-p38 (P = 0.017), and p-ERK (P = 0.041) retained their predictive value. CONCLUSIONS: Our results present the first evidence of in vivo involvement of MAPKs in the clinical course of ovarian cancer and the possible effect of chemotherapy on intracellular signaling in this disease. The improved prognosis associated with expression and phosphorylation of all three mitogen activated protein kinase families highlights the unique properties of cancer cells in effusions and may expand our understanding of the biology of ovarian carcinoma at this site, possibly affecting treatment strategies for this malignancy. PMID- 14529678 TI - Apoptotic pathways of epothilone BMS 310705. AB - OBJECTIVE: BMS 310705 is a novel water-soluble analog of epothilone B currently in phase I clinical evaluation in the treatment of malignancies such as ovarian, renal, bladder, and lung carcinoma. Using an early passage cell culture model derived from the ascites of a patient clinically refractory to platinum/paclitaxel therapy, we evaluated the pathway of caspase-mediated apoptosis. METHODS: Cells were treated for 1 h and subsequently evaluated for apoptosis, survival, and caspase activity. Apoptosis was determined by fluorescent microscopy. Caspase-3, -8, and -9 activities were determined by fluorometry using target tetrapeptide substrates. Mitochondrial release of cytochrome c was determined by immunoblot analysis. RESULTS: After treatment with BMS 310705, apoptosis was confirmed in >25% of cells at 24 h. Survival was significantly lower (P < 0.02) in cells treated with 0.05 micro M BMS 310705 vs paclitaxel. Analysis revealed an increase of caspase-9 and -3 activity; no caspase -8 activity was observed. Release of cytochrome c was detected at 12 h following treatment. SN-38 and topotecan failed to induce apoptosis. CONCLUSIONS: BMS 310705 induces significant apoptosis, decreases survival, and utilizes the mitochondrial-mediated pathway for apoptosis in this model. PMID- 14529679 TI - Laparoscopic port-site metastases: etiology and prevention. AB - OBJECTIVE: The purpose of this article is to summarize current hypotheses for the possible sources of laparoscopic port-site metastasis, to review the results of experimental models that support such hypotheses, and to discuss the potential options for preventing these metastases. METHODS: We performed a Medline search to identify in vitro and in vivo studies and clinical trials that analyzed port site metastases associated with laparoscopic surgery. We report the incidence of port-site metastases and causative factors associated with this condition. RESULTS: The estimated incidence of port-site metastases in all patients undergoing laparoscopic surgery for malignant disease is approximately 1-2%. Multiple factors are associated with this complication. Among the most common proposed etiologies are the wound implantations caused by the surgical technique and instrumentation; the leakage of insufflation gas through the ports, known as the "chimney effect"; and the impact of pneumoperitoneum on local immune reactions. Several preventive measures, have been suggested, including careful patient selection, lavage of the peritoneal cavity as well as of the port wounds with cytotoxic agents, and modifications of surgical technique. CONCLUSIONS: Only through the results of well-conducted large multi-institutional prospective randomized trials will we learn not only the true incidence of port-site metastases, but also the potential factors that lead to the occurrence of this complication. PMID- 14529680 TI - Comparison of neutropenia in a randomized, crossover trial of 3-, 6-, and 24-h infusions of paclitaxel. AB - OBJECTIVE: Paclitaxel is most commonly infused over 3 hs rather than the original schedule of 24 h as the briefer infusion duration results in greater convenience, similar efficacy, significantly less myelosuppression, and less cost. While differences in toxicity between 3- and 24-h infusions are well described, there is little information about the effect of modest prolongation of infusion duration, which is often employed in patients who develop hypersensitivity reactions. To assess whether prolonging a 3-h infusion significantly increases the degree of neutropenia, we reviewed our data from a randomized, crossover trial of 3-h versus 6-h versus 24-h regimens of paclitaxel. METHODS: Results from 12 patients who were randomized to receive one 3-h, one 6-h and one 24-h infusion of paclitaxel in varied sequences during their first three cycles of treatment were analysed. The blood counts were monitored closely throughout each cycle of treatment. RESULTS: Crossover trial methodology was used to assess the differences in the degree of neutropenia caused by the three different infusion regimens. The 24-h infusion regimen resulted in significantly worse neutropenia than the 3- or 6-h infusion regimens. There was no statistically significant difference between the 3- and 6-h infusion regimens with respect to all endpoints. The estimated mean difference in the duration of grade 3 or 4 neutropenia between the 3- and 6-h infusion regimens (6 h - 3-h) was 1.1 day (95% CI: -0.9, 3.2), and for grade 4 neutropenia, the estimated mean difference in the duration was 0.8 day (95% CI: -0.4, 2.0). CONCLUSIONS: Increasing the duration of paclitaxel infusions from 3 to 6 h does not result in a statistically significant increase in the degree of neutropenia. Any additional neutropenia is likely to be of brief duration. PMID- 14529681 TI - Tumor cytosol dipeptidyl peptidase III activity is increased with histological aggressiveness of ovarian primary carcinomas. AB - OBJECTIVE: Proteolytic enzymes have been implicated in the progression of various human malignancies, including ovarian cancer. The enhanced expression of dipeptidyl peptidase III (DPP III) was found in endometrial carcinomas of various histological types and grade. The aim of this study was to assess activity of DPP III in ovarian tissue specimens and to correlate it with clinico-pathological data. METHODS: DPP III hydrolytic activity toward Arg-Arg-2-naphthylamide was determined in 108 ovarian tissue cytosol specimens of 79 patients. The data obtained for 41 ovarian primary carcinoma specimens were stratified according to clinical stage, histological grade and type, and age of the patients. RESULTS: Median DPP III activity expressed as milliunits per milligram protein was 6 in normal ovarian tissues (n = 29), 6.5 in benign ovarian tumors (n = 19), 19.5 in primary ovarian carcinomas (n = 41), 12.5 in nonepithelial primary ovarian tumors (n = 7), and 22.1 in metastatic ovarian malignancies (n = 12). A significant rise in median DPP III specific activity was observed in malignant ovarian tumors (of epithelial, nonepithelial, and metastatic origin), but not in benign ovarian tumors, compared to the activity in normal tissue. A significant difference of DPP III expression was found between the group of normal tissues and tumors of clinical stage I and II, of grade 2 and 3, of serous and mucinous histologic type. CONCLUSIONS: DPP III activity of benign ovarian tumors equaled that in normal ovarian tissue. In malignant neoplasms of the ovary it increased with growing histologic grade. PMID- 14529682 TI - Expression of p16 INK4A in Papanicolaou smears containing atypical squamous cells of undetermined significance from the uterine cervix. AB - OBJECTIVE: This study aimed to verify one of the major diagnostic dilemmas in routine Papanicolaou (Pap) smears. Atypical squamous cells of undetermined significance (ASCUS) were tested immunocytochemically for p16(INK4A) (p16), and results were correlated with follow-up biopsies for more accurate diagnoses. METHODS: The study included 66 Pap smears of ASCUS diagnostic categories, all of which were correlated histologically. The cytological diagnoses of ASCUS were further classified cytologically according to the 2001 Bethesda System, as "atypical squamous cells of undetermined significance (ASC-US)" or "atypical squamous cells which cannot exclude HSIL (ASC-H)." All Pap smears were decolorized and immunostained with the primary anti-p16 antibody, clone E6H4. Immunoreactivity for p16 was correlated with histological sections (which were also immunostained for comparison) in a semiblind fashion. RESULTS: Of the 66 smears containing ASCUS, 47 (71%) were reclassified as ASC-US and 19 (29%) as ASC H. Follow-up biopsies revealed that 21 (32%) cervices had no obvious abnormalities but only reactive changes. A significant proportion of histological diagnoses were CIN1/LSIL (24 cases, 36%), CIN2 or 3/HSIL (17 cases, 26%), squamous cell carcinoma (two cases, 3%), or AIS/adenocarcinoma (two cases, 3%). The p16 immunocytochemical stain was reactive in 40 (60.6%) of 66 smears: either weakly/sporadically (18 cases, 45%) or strongly positive (22 cases, 55%). Conversely, 26 (39.4%) of the smears were negative for p16 and displayed predominantly reactive changes. However, five cases of LSIL and one of HSIL were negative for p16. From the results of p16 immunoreactivity of atypical cells for detection of biopsy-proved significant lesions (HSIL or higher), this analysis was highly sensitive (sensitivity, 95%) and specific (specificity, 96%) and had favorable positive (91%) and negative (98%) predictive values. CONCLUSIONS: On the basis of both morphological and immunostaining patterns, there is a clear association between strong p16 immunostaining of atypical cells in the smears and the presence of significant lesions in the cervix, except in two patients. Similarly, there is a clear association between the lack of p16 expression and the absence of cervical lesions. The p16 immunocytochemical stain can be applied successfully to conventional Pap smears and may serve as a useful biomarker in ASCUS-containing smear diagnoses. This may offer a more objective parameter to help clarify this ambiguous area in gynecological cytopathology. PMID- 14529683 TI - Incidence of lymph node and ovarian metastases in leiomyosarcoma of the uterus. AB - OBJECTIVE: The purpose of this study was to determine the incidence of lymph node and ovarian metastases in newly diagnosed uterine leiomyosarcoma (LMS), and to describe possible predictive factors. METHODS: We used our prospectively acquired databases to identify 275 consecutive patients with uterine LMS treated from 7/82 to 12/01. Patients were included if there was clear documentation of lymph nodes and/or ovarian tissue in the pathologic reports. Clinical data were extracted from electronic medical records. Statistical analysis using the Fisher exact test was used to determine prognostic factors. RESULTS: There were 108 patients (39.2%) identified in whom an oophorectomy and 37 patients (13.5%) in whom lymph node sampling was performed as part of the initial surgical management of uterine LMS. Bilateral oophorectomy was performed in 102 (94.4%) of the 108 patients. The median numbers of pelvic, para-aortic, and total lymph nodes acquired were 5 (range, 1-27), 3 (range, 1-9), and 6 (range, 1-34), respectively. Ovarian metastases were found in 4 (3.9%) out of 108 patients. Two (2.8%) of the 71 patients with disease confined to the uterus and/or cervix (stage I/II) and 2 (5.4%) of the 37 patients with gross extrauterine disease had ovarian metastases (P = 0.43). Positive lymph nodes were seen in 3 (8.1%) of 37 patients. No patients with stage I/II disease had positive lymph nodes (P = 0.015). None of the factors analyzed predicted for metastases to the ovary. Only the presence or absence of gross extrauterine disease correlated with lymph node metastasis. In addition, all three of these cases had clinically suspicious (enlarged) lymph nodes. CONCLUSIONS: The incidence of ovarian and lymph node metastases in uterine LMS is very low and is most commonly associated with extrauterine disease. Lymph node dissection for uterine LMS should be reserved for patients with clinically suspicious nodes. PMID- 14529684 TI - The use of music to reduce anxiety for patients undergoing colposcopy: a randomized trial. AB - OBJECTIVE: The goal of this work was to investigate the impact of music on women's anxiety and perceived pain during colposcopy examination. METHODS: This was a prospective randomized study. Two hundred and twenty women referred for colposcopy for the first time were recruited. They were randomized to either the music or no-music group. Before colposcopy examination, each subject completed a Chinese version of the state anxiety questionnaire (STAI) and assessed the anticipated pain for colposcopy with a visual analog scale (VAS). Slow-rhythm music was played during colposcopy examination in the music group. Subjects in the no-music group were examined in the same setting without music. After colposcopy, each subject completed the STAI form again and assessed their pain during examination by the VAS. RESULTS: Women in the music group experienced significantly less pain (mean VAS 3.32 [95% CI 2.86-3.78] vs 5.03 [4.54-5.52], P<0.001) and lower anxiety (mean STAI 39.36 [95% CI 37.33-41.39] vs 44.16 [41.82 46.49], P = 0.002) during colposcopy examination than women in the no-music group. On linear regression analysis, the factors significantly affecting anxiety during colposcopy were anxiety score at enrollment, pain score during colposcopy, and whether or not the women had listened to music during the colposcopy examination. The factors significantly affecting the pain scores were whether the women had listened to music during the procedure and the final anxiety scores. CONCLUSIONS: Music is a simple, inexpensive, and easily used strategy to minimize anxiety and pain during colposcopy examination. PMID- 14529685 TI - Molecular assessment of depth of myometrial invasion in stage I endometrial cancer: a model based on K-ras mutation analysis. AB - INTRODUCTION: Overall nearly 20% of endometrial cancer (EC) patients die of the disease and over half of these had initially presented with clinical stage I disease. There is a strong correlation between disease mortality and depth of myometrial invasion. Current assessment of depth of invasion relies on light microscopy. Tumor cells can evade detection by light microscopy if they are vastly outnumbered by myometrial cells. Molecular techniques have a great potential in the detection of such isolated cells. OBJECTIVE: The objective was to develop a model for the application of molecular techniques to advance the assessment of risk status in patients with clinical stage I EC. METHODS: The study sample included 21 stage I ECs with a documented K-ras mutation from two series of 96 and 106 ECs from the United Kingdom and Norway, respectively. K-ras was documented using heteroduplex mobility analysis and amplified created restriction site, followed by sequencing to identify the specific base substitution at codon 12 and 13 of K-ras oncogene. For each case with a K-ras mutation, a modified mutant allele-specific amplification technique was carried out on a series of tissue strips microdissected at increasing depths from the myometrium underlying tumor. The microdissected myometrium had been previously examined histologically for absence of infiltrating tumor cells on light microscopy. Presence of K-ras mutations was used to identify the tumor cells within the histologically normal myometrium. Correlations between submicroscopic myometrial tumor cell infiltration and clinicopathological factors were studied. RESULTS: Of 21 cases with K-ras mutation, 6 cases (28%) showed molecular evidence of tumor cell infiltration beyond the histological boundary. The depth of submicroscopic myometrial infiltration was found to be variable. The staging of the tumors would have changed in 3 cases (14%) if tumor cells been detected histologically. A borderline significant correlation between presence of submicroscopic myometrial invasion and depth of myometrial invasion was noted (P = 0.053). The recurrence rate and survival of patients without submicroscopic invasion were better than those with, although it did not reach statistical significance (recurrence rate P = 0.13, recurrence free survival P = 0.14, cause specific survival P = 0.12, and total survival P = 0.2). CONCLUSIONS: Molecular assessment of depth of myometrial invasion using K-ras mutation is feasible and may add information to conventional light microscopy. Further prospective studies are required to define the clinical significance of this technology. PMID- 14529686 TI - [18F]fluorodeoxyglucose positron emission tomography as a useful indicator of metastatic gestational trophoblastic tumor: preliminary results in three patients. AB - OBJECTIVE: The goal of this study was to evaluate the usefulness of positron emission tomography with [(18)F]fluorodeoxyglucose ([(18)F]FDG-PET) in detecting metastases in patients with gestational trophoblastic tumor (GTTs). METHODS: A retrospective study was conducted on three patients with GTTs who had been studied with [(18)F]FDG-PET and computed tomography (CT) after an increase in human chorionic beta-gonadotropin (betahCG) serum levels. PET scans were performed with a multiring whole-body positron emission tomograph 45 min after an intravenous bolus injection of [(18)F]FDG ( approximately 5.2 MBq/kg). CT studies were obtained on a spiral scanner prior and after administration of intravenous iodinated contrast material. Within a week of CT and [(18)F]FDG-PET studies, the patients underwent surgical procedures for histological diagnosis. RESULTS: In one patient, a lung lesion positive for neoplastic tissue with [(18)F]FDG-PET and negative with CT was confirmed to be a GTT metastasis at histology. In another patient, [(18)F]FDG-PET was negative, while CT was positive for the presence of lung metastasis; no viable tumor tissue was found at histological analysis. The remaining patient had a positive [(18)F]FDG-PET and CT study for lung metastasis; this was confirmed at histological analysis. In the same patient, both [(18)F]FDG PET and CT depicted the presence of a liver lesion. Necrotic lesion regression after treatment was clearly documented with [(18)F]FDG-PET only. CONCLUSIONS: Our preliminary results suggest that [(18)F]FDG-PET may be useful for the assessment of metastatic disease in patients with GTTs. PMID- 14529687 TI - Selection of HER-2/neu-positive tumor cells in early stage cervical cancer: implications for Herceptin-mediated therapy. AB - OBJECTIVE: The purpose of this study was to compare HER-2/neu expression on biopsies obtained from early stage cervical cancer, primary cell lines established therefrom, established cervical cancer cell lines, and metastatic or recurrent sites of disease; and to evaluate the sensitivity of primary cervical cancer cell lines to treatment with a humanized MAb against HER-2/neu (Herceptin). METHODS: Surface HER-2/neu expression on 18 cervical cancer cell lines was compared to HER-2/neu detection by immunohistochemistry on biopsies obtained from the original tumors (10 patients) and sites of recurrence (2 patients). Primary cell lines were tested for sensitivity to Herceptin-mediated antibody-dependent cellular cytotoxicity (ADCC) and sensitivity to Herceptin mediated inhibition of proliferation. RESULTS: Nine out of 10 primary (90%) and 8 out of 8 (100%) established cervical cancer cell lines expressed HER-2/neu by flow cytometry. Surprisingly, all HER-2/neu-positive primary cell lines were derived from tumor biopsies that scored negative (i.e., 0 to 1+) for HER-2/neu expression by immunohistochemistry. Heavy staining for HER-2/neu (i.e., 3+) was found in the recurrent/metastatic lesions of the two relapsed patients. Importantly, all HER-2/neu-positive primary cell lines were highly sensitive to Herceptin-mediated ADCC, and their proliferation was also significantly inhibited by Herceptin. A significant enhancement of Herceptin-mediated ADCC was demonstrated when effector cells were exposed to low doses of IL-2 in vitro. CONCLUSIONS: Early stage cervical cancer may develop a population of HER-2/neu positive cells with a selective growth advantage over HER-2/neu-negative cells. Therapy which targets HER-2/neu may be more effective in patients with cervical cancer than indicated by the commonly low expression of HER-2/neu in tumors removed at the time of primary treatment. PMID- 14529688 TI - Vulvar Langerhans cell histiocytosis: a case report and review of the literature. AB - BACKGROUND: Langerhans cell histiocytosis (LCH) of the female genital tract is rare. Only, seven cases of primary vulvar LCH have been previously reported in the medical literature. We describe an additional case of LCH in which the disease was confined to the vulva. CASE: A 33-year-old gravida 0, para 0 Ethiopian woman presented with a nodular lesion on her left vulva. The lesion was biopsied, and the results were consistent with LCH. A metastatic workup did not reveal any evidence of disease beyond the vulva. The patient was initially treated with radiotherapy to the vulva. She was diagnosed with recurrent disease in the vulva 21 months after the completion of radiotherapy. At that time, she underwent a wide local excision. Five months later, we found a lesion on her right labium majus that was consistent with a recurrence. The patient's vulva was treated with a higher dose of radiotherapy than it had been the first time. Six months later the patient again experienced a local recurrence. She underwent a wide radical vulvar excision of diffuse bilateral lesions and was free of disease for approximately 3 months, after which she experienced another recurrence and underwent treatment with thalidomide. Within 2 months of starting thalidomide therapy, the patient experienced resolution of her symptoms and of her vulvar lesions. CONCLUSIONS: Primary LCH of the vulva is very rare. Its etiology and pathophysiology, as well as the most effective modes of therapy, remain elusive. We propose that thalidomide is a useful alternative for patients with this disease. PMID- 14529689 TI - Pelvic primitive neuroectodermal tumor associated with a cluster of small round cell tumors: case report and review of current literature. AB - BACKGROUND: Peripheral primitive neuroectodermal tumor (pPNET) is aggressive and rare, comprising 1% of soft tissue sarcomas. Involvement of the reproductive tract is unusual. CASE: A 35-year-old woman had a pelvic mass and omental cake. Frozen-section examination at laparotomy revealed small round cell tumor confirmed as pPNET. Chemotherapy with vincristine, doxorubicin, and cyclophosphamide alternating with ifosfamide and etoposide with mesna yielded complete response. The patient's mother died of a similar tumor at age 52 years, and the patient's husband had adult Ewing sarcoma, constituting an unusual cluster of related tumors. CONCLUSIONS: Genetic recombination resulting in a chimeric transcript of the Ewing sarcoma gene and the Friend leukemia virus integration site is characteristic of these tumors. Surgical resection and multiagent chemotherapy may enhance survival. PMID- 14529690 TI - Direct laparoscopic venous sampling to diagnose a small Sertoli-Leydig tumor. AB - BACKGROUND: Sertoli-Leydig cell tumors (SLCT) constitute only 0.5% of all primary ovarian neoplasms. We report a unique diagnostic method (selective laparoscopic venous sampling) and a rare case of a contralateral second primary tumor. CASE: A 14-year-old female presented with hyperandrogenic complaints and an increased serum testosterone. Ovarian origin was confirmed by direct laparoscopic ovarian blood sampling. A right salpingo-oophorectomy was performed. The pathological diagnosis was SLCT of intermediate differentiation. Three years later, the patient presented again with an increased serum testosterone. A solid tumor in the left ovary was excised. The pathology was SLCT of intermediate differentiation. The patient remains disease-free. CONCLUSIONS: Direct laparoscopic venous sampling is used to diagnose a small SLCT in a teenage patient. PMID- 14529691 TI - High dose rate intraoperative radiotherapy for recurrent cervical cancer and nodal disease. AB - BACKGROUND: Patients with pelvic sidewall recurrences of cervical cancer have a dismal prognosis. Intraoperative radiation therapy (IORT) has demonstrated encouraging results. Patients have traditionally been excluded from IORT if they had distant metastases. CASE: A patient underwent radical tumor resection and high dose rate (HDR) IORT for a pelvic sidewall recurrence of cervical cancer. She also had metastatic disease in a para-aortic node. The patient has been followed for >20 months with no evidence of disease recurrence. CONCLUSIONS: HDR IORT may be offered to select patients with recurrent cervical cancer and isolated metastatic para-aortic lymph nodes. PMID- 14529692 TI - The utility of gastrojejunostomy in secondary cytoreduction and palliation of proximal intestinal obstruction in recurrent ovarian cancer. AB - BACKGROUND: Gastrointestinal obstruction is a common complication of recurrent ovarian cancer. Proximal intestinal obstruction, at the level of the duodenum or proximal jejunum, can result from bulky intraperitoneal or retroperitoneal disease. Classic management has been palliation of symptoms with a gastrostomy or jejunostomy tube. CASE: We describe a series of four patients with recurrent ovarian carcinoma and proximal intestinal obstructions treated with a bypass stapled side-to-side gastrojejunostomy at the time of secondary cytoreduction or surgical palliation. The clinical history, preoperative evaluation, surgical technique, and outcomes of each patient are reviewed. CONCLUSIONS: Gastrojejunostomy may offer patients with ovarian cancer and a proximal intestinal obstruction symptomatic relief and an opportunity for resumption of enteral feedings. PMID- 14529693 TI - Endometrial stromal sarcoma presenting as postpartum haemorrhage: report of a case with a sole t(10;17)(q22;p13) translocation. AB - BACKGROUND: Although the clinical picture of endometrial stromal sarcoma (ESS) is variable, it was never reported to present as a postpartum hemorrhage. In addition, ESS is a tumor type of which, due to its rarity, little is known regarding chemosensitivity and genetic changes. CASE: A 28-year-old woman complaining of persistent postpartum bleeding was referred to our hospital, where she was diagnosed with ESS. At laparotomy, the invasion of nervous and vascular pelvic structures rendered her inoperable, and chemotherapy (doxorubicin 50 mg/m(2) for 15 min; ifosfamide 5 g/m(2)/24 h; mesna 5 g/m(2), every 3 weeks) was initiated. The ESS appeared to be chemosensitive because after three treatment cycles the tumor iliac metastase significantly decreased in volume and became surgically removable. Chemosensitivity was confirmed microscopically. Three additional courses of chemotherapy and pelvic irradiation were administered. Cytogenetic evaluation of both the primary as well as the metastatic lesions revealed a t(10;17)(q22;p13) as the sole cytogenetic abnormality. CONCLUSIONS: Three interesting features of this particular case put ESS in a new perspective. First, the fundal ESS permitted normal conception and pregnancy but caused a postpartum haemorrhage. Second, the ESS was clearly chemosensitive. Third, we report a novel cytogenetic aberration in ESS, the molecular characterization of which might lead to the identification of the deregulated pathway(s) triggering tumor development in ESS. PMID- 14529694 TI - Chemopersistent early recurrence in the perineal tear scar of an intrapartally diagnosed cervical cancer. AB - BACKGROUND: Cervical cancer is the most frequent cancer occurring in pregnancy. Apart from some general recommendations, there is no standardized consensus for the management of this cancer during pregnancy. Individual case reports do exist on intrapartal cancer diagnosis and risks. CASE: We describe the case of an early recurrence in the perineal tear scar of an intrapartally diagnosed cervical cancer, which had developed despite combined neoadjuvant and adjuvant chemotherapy. CONCLUSIONS: Although iatrogenic tumor cell spreading in the vagina is rare, this oncological risk is easy to avoid. It should be considered in the peripartal management of patients suffering from cervical cancer. Our case report underlines the limits of chemotherapy in the treatment of tumor residue and points out the problem of cervical cancer cell spreading during invasive procedures. PMID- 14529695 TI - Infrared spectral features of exfoliated cervical cells, cervical adenocarcinoma tissue, and an adenocarcinoma cell line (SiSo). By Neviliappan S, et al. PMID- 14529697 TI - The etiology of social phobia: toward a developmental profile. AB - Social phobia is an extremely disruptive and distressing anxiety disorder that can impact on many areas of an individual's life. Yet, despite the fact that lifetime prevalence rates are relatively high, its etiology is still poorly understood. The aim of this review is to draw together findings from the broad base of nonclinical literature associated with behavioral inhibition (BI), shyness, social anxiety, and passive-anxious withdrawal and to compare these findings with those from the limited number of clinical studies with social phobics. Such comparison is not unproblematic due to conceptual differences between terms used and methodological divergence; these issues are discussed in some detail. The consonance of findings, however, suggests a viable profile for the developmental course of social phobia. This profile incorporates temperament variables, behavioral motivational, parenting styles, peer relationships, and internalization problems. Finally, specific suggestions for future research are offered. PMID- 14529698 TI - Anxiety and depression in juvenile diabetes: a critical review. AB - A critical examination of the recent literature on anxiety and depression in juvenile diabetes is presented. The objectives of this review are: (1) to determine the general association of psychological factors, especially anxiety and depression, with diabetes, (2) to examine the specific association of anxiety and depression with metabolic control, and (3) to propose methodological changes that are needed to advance future research in this field. The major conclusions of this review support the notion of a general association of psychological disorders with juvenile diabetes. However, while anxiety and depression appear to play an important and complex role in determining adaptation to the disease, their relationship to metabolic control does not yet appear clear. Additional prospective and controlled studies as well as multivariate models of chronic disease are now necessary to more fully understand the etiology and impact of these disorders in the adolescent population. PMID- 14529699 TI - Searching for the attention deficit in attention deficit hyperactivity disorder: the case of visuospatial orienting. AB - We review all 14 extant studies of covert visuospatial attention in attention deficit hyperactivity disorder (ADHD) (total N=248). Metaanalysis showed that intriguing but isolated findings of alerting or posterior disengage deficits were too small to reliably detect with the sample sizes typically employed. Posterior move and engage operations and the vigilance sustained attention process were normal in ADHD. For exogenous cues, effect sizes for group differences were homogeneously small across all repeated-measures conditions, as were calculations of cost, benefit, and validity effects. For endogenous cues, effect sizes were heterogeneous; however, calculations of cost, benefit, and validity effects were small and homogenous. The most parsimonious conclusion may be that ADHD is not characterized by significant visual orienting dysfunction, but questions remain about the extent of anterior lateralized effects in the combined subtype and about attentional functioning in the inattentive subtype. PMID- 14529700 TI - Anxiety and depression: why and how to measure their separate effects. AB - It is well recognized that depression and anxiety are closely associated, but nonetheless, they may be associated with distinct causes and consequences. For example, anxiety and depression are associated with different effects on information processing. This paper argues that experiments should not study anxiety or depression in isolation, but should measure both variables. It is argued that this methodological step is both important and commonly overlooked. Even when both depression and anxiety are measured, methodological difficulties can confuse their effects. Common difficulties in choice of measures and in participant selection criteria are discussed, and recommendations are made for overcoming them. The argument is made drawing from illustrations within the experimental cognitive literature, but conclusions and recommendations are equally applicable outside this area. PMID- 14529701 TI - Attributions and expressed emotion: a review. AB - This review examines the contribution of an attribution-based framework to our understanding of both expressed emotion (EE) and the process of patient relapse. It considers all published studies that have assessed the attributions of high- and low-EE relatives and carers. Across these studies, a great deal of consistency is apparent. Critical relatives are more likely to hold patients responsible for their difficulties. On the other hand, attributions made by emotionally overinvolved relatives are similar to the attributions made by those who are low EE. In reviewing the association between attributions and patient relapse, it is concluded that carers' beliefs may play a role in the relapse process in a variety of ways. These include mediating controlling behavior, which may serve to increase patient stress or decrease patients' sense of self-worth. A major limitation of research in this area is the cross-sectional and correlational nature of the data. This makes it impossible to draw conclusions about directions of effect in the association of EE and attributions. Nonetheless, an argument can be made for the development and evaluation of interventions designed to help relatives adopt less blaming and more flexible beliefs about the nature of patients' problems. Particularly helpful in this regard may be interventions that use verbal and behavioral reattribution techniques similar to those described in the cognitive behavioral treatment of emotional disorders. PMID- 14529702 TI - Social stress and T cell maturation in male rats: transient and persistent alterations in thymic function. AB - The present study demonstrates that social stress in subordinate male rats resulting from a 24-h confrontation with a dominant male opponent caused not only transient but also persistent alterations in the function and the cellular composition of the thymus. The investigation of regulation processes maintaining tissue homeostasis in the thymus showed a significant increase in apoptosis, a decrease in cellular proliferation and a decrease in thymic export as indicated by a reduction in the number of recent thymic emigrants (RTE) in the peripheral blood. The proliferative response of mature thymocytes to Concanavalin A (ConA) was also significantly reduced in socially defeated rats. Adrenalectomy (ADX) completely abolished the stress-associated alterations in thymic function whereas a decrease in total thymocyte numbers was still evident in defeated animals treated with the glucocorticoid type-II receptor antagonist RU486. The significant reduction in total thymocytes and the diminished proliferative response to ConA persisted for at least seven days after stressor cessation. An enhanced proliferative activity at this point of time may represent a beginning regeneration of thymic tissue after stress. Taken together, our studies show that social stress profoundly disturbs tissue homeostasis in the thymus and suggest that adrenal-derived hormones play a central role in mediating this effect. The strong positive relationship between total thymocyte numbers and peripheral RTE numbers and the long-lasting atrophy of the thymus in defeated animals may suggest negative consequences for the heterogeneity of naive T cells in the peripheral T cell pool after social stress. PMID- 14529703 TI - The influence of photoperiod and sex on lipopolysaccharide-induced hypoactivity and behavioral tolerance development in meadow voles (Microtus pennsylvanicus). AB - Lipopolysaccharide (LPS), the minimal immunogenic component of Gram-negative bacteria, is released during infection and causes a variety of sickness behaviors including decreased locomotor activity. This study considered how photoperiod and sex influence the effects of LPS in the meadow vole, Microtus pennsylvanicus. Male and female voles were housed under either reproductively stimulatory (long day: 16 h) or inhibitory (short day: 8 h) photoperiods. On Days 1 and 8, voles were injected with LPS (200 microg/kg, i.p.) or saline vehicle and locomotor activity was assessed 2 h later in an automated open field for 1 h. The first exposure to LPS caused significant decrements in locomotor activity in all LPS treated groups, regardless of photoperiod or sex. On Day 8, both short day males and females exhibited behavioral tolerance to LPS, no longer displaying significant activity decrements. In contrast, long day females reinjected with LPS on Day 8 still exhibited significant hypoactivity on all locomotor measures. Similarly, long day males also appeared to exhibit a sustained expression of sickness behaviors on Day 8. In long day females, higher circulating progesterone levels were associated with an attenuated rate of tolerance formation to LPS. The present findings support the winter immunoenhancement hypothesis, which states that small mammals which undergo severe seasonal fluctuations undergo compromised immune functioning during the breeding season, and further indicate a potential role for progesterone in modulating these seasonal immune fluctuations in females. PMID- 14529704 TI - Glucocorticoids are involved in the long-term effects of a single immobilization stress on the hypothalamic-pituitary-adrenal axis. AB - We have previously observed that a single exposure to a severe stressor such as immobilization (IMO) induces long-lasting desensitization of the responsiveness of the hypothalamic-pituitary-adrenal (HPA) axis to the same stressor that enhances rather than dissipates with time (days). As this desensitization of the HPA axis was not observed in response to a novel stressor, we suggested this might be a particular type of learning linked to severe stressful situations. Taking into account the evidence that glucocorticoids are involved in learning and memory, the present study addresses the role of glucocorticoids in the induction of long-term effects of an acute exposure to IMO. Three different experimental approaches were used: (i) blockade of stress-induced corticosterone release by using adrenalectomized rats supplemented with a low dose of corticosterone in the drinking saline (ADX+B); (ii) blockade of corticosterone synthesis during the first exposure to IMO with the 11-beta-hydroxylase inhibitor metyrapone (200 mg/kg); and (iii) administration of the glucocorticoid receptor antagonist RU486 (100 mg/kg). Previous exposure to IMO resulted in an enhanced post-stress recovery of the HPA response to the same stressor 1 week later. These long-term effects of IMO were blocked in ADX+B rats, were partially reduced in metyrapone-treated rats and only modestly affected by RU486 administration. These data suggest that glucocorticoids play a partial role in the induction of long term effects of IMO on the HPA responsiveness to the same stressor, although the weak effect of RU486 suggests that non-classical corticosteroid receptors may be involved. The role of glucocorticoids in the expression of the phenomenon is suggested by the full blockade of the phenomenon in ADX+B rats, but further studies are needed. As blockade of corticosterone synthesis only partially blunted the long-term effect of IMO, it appears that full induction of the long term effects of acute exposure to IMO on the HPA axis is only achieved by the concerted action of several endocrine (or neurochemical) factors. PMID- 14529705 TI - Spatial abilities following prenatal androgen abnormality: targeting and mental rotations performance in individuals with congenital adrenal hyperplasia. AB - In most mammals, behaviors that show sex differences are influenced by androgen during early life. In the current study, the hypothesis that androgen influences the development of human spatial abilities was investigated. Participants included 40 females and 29 males with congenital adrenal hyperplasia (CAH), a genetic disorder that causes overproduction of adrenal androgens beginning prenatally, and 29 unaffected female and 30 unaffected male relatives of individuals with CAH. Participants ranged in age from 12-45 years. Measures of spatial abilities included two mental rotations tasks and two targeting tasks, all of which showed large sex differences favoring males in the unaffected relative controls. Females with CAH (exposed to higher than normal levels of androgen prenatally) performed better than unaffected females on the targeting tasks, and resembled unaffected males and males with CAH in this respect. However, females with CAH did not perform better than unaffected females on the measures of mental rotations abilities. Males with CAH showed unaltered performance on the targeting tasks, and impaired performance on the mental rotations tasks. Results are discussed in terms of differences in experiential and hormonal contributions to different spatial abilities, as well as in terms of possible differences in critical periods for hormonal influences on targeting versus mental rotations abilities. Specifically, we speculate that, although androgen may influence targeting abilities prenatally, if hormones influence the development of mental rotations ability, they do so at some other time, perhaps during the first six months of postnatal life. PMID- 14529706 TI - Photoperiod affects the expression of sex and species differences in leukocyte number and leukocyte trafficking in congeneric hamsters. AB - Sex differences in immune function are well documented. These sex differences may be modulated by social and environmental factors. Individuals of polygynous species generally exhibit more pronounced sex differences in immune parameters than individuals of monogamous species, often displaying an energetic trade-off between enhanced immunity and high mating success. During winter, animals contend with environmental conditions (e.g. low temperatures and decreased food availability) that evoke energetic-stress responses; many mammals restrict reproduction in response to photoperiod as part of an annual winter coping strategy. To test the hypothesis that extant sex and species differences in immune surveillance may be modulated by photoperiod, we examined leukocyte numbers in males and females of two closely related hamster species (Phodopus). As predicted, uniparental P. sungorus exhibited a robust sex difference, with total white blood cells, total lymphocytes, T cells, and B cells higher in females than males, during long days when reproduction occurs, but not during short days when reproduction usually stops. In contrast, biparental male and female P. campbelli exhibited comparable leukocyte numbers during both long and short days. To study sex differences in stress responses, we also examined immune cell trafficking in response to an acute (2 h) restraint stressor. During stressful challenges, it appears beneficial for immune cells to exit the blood and move to primary immune defense areas such as the skin, in preparation for potential injury or infection. Acute stress moved lymphocytes and monocytes out of the blood in all animals. Blood cortisol concentrations were increased in P. sungorus females compared to males at baseline (52%) and in response to restraint stress (38%), but only in long days. P. campbelli males and females exhibited comparable blood cortisol and stress responses during both long and short days. Our results suggest that interactions among social factors and the environment play a significant role in modulating sex and seasonal alterations in leukocyte numbers and stress responses. PMID- 14529707 TI - Food preferences, testosterone concentrations and offspring sex ratios of female mice. PMID- 14529709 TI - Regulation of AMPA receptor dephosphorylation by glutamate receptor agonists. AB - Phosphorylation of the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor subunit GluR1 at Ser(845) enhances AMPA channel activity. This study demonstrates that Ser(845) is rapidly dephosphorylated upon AMPA receptor activation in nucleus accumbens slices. AMPA-induced dephosphorylation at Ser(845) was blocked by CNQX, an AMPA receptor antagonist, by nifedipine, an L type Ca(2+) channel antagonist, or by cyclosporin A, a calcineurin inhibitor. N methyl-D-aspartate (NMDA) treatment also decreased phosphorylation of Ser(845), an effect that was blocked by MK-801, an NMDA receptor antagonist, but not by nifedipine. Accumbens neurons are enriched for dopamine- and cyclic AMP (cAMP) regulated phosphoprotein, Mr 32,000 (DARPP-32), a potent inhibitor of protein phosphatase 1 (PP1) when phosphorylated by PKA (at Thr(34)). We tested the hypothesis that the AMPA/KA or NMDA-stimulated dephosphorylation of DARPP-32 via calcineurin, leading to increased PP1 activity and dephosphorylation of GluR1. AMPA or NMDA treatment decreased phospho-Thr(34)-DARPP-32 levels, effects that were blocked by receptor antagonists, or cyclosporin A. However, dephosphorylation of Ser(845) mediated by AMPA or NMDA receptors was unaffected in DARPP-32/inhibitor-1 knockout mice. These data suggest that AMPA- or NMDA induced dephosphorylation of GluR1 at Ser(845) occurs by a mechanism that is independent of DARPP-32 and PP1, but involves activation of calcineurin. Thus, Ca(2+)-dependent dephosphorylation of GluR1 may serve as a negative feedback mechanism for the regulation of AMPA receptor activity in neurons. PMID- 14529710 TI - An interaction domain in Slob necessary for its binding to the slowpoke calcium dependent potassium channel. AB - Slob modulates the activity of the Drosophila Slowpoke calcium-dependent potassium channel (dSlo) via its direct binding to the channel. To characterize the molecular detail of the protein-protein interaction between Slob and dSlo, we constructed a series of Slob mutants that are progressively truncated at either the carboxyl or amino terminal end, and examined the binding of these Slob mutants to dSlo using a co-immunoprecipitation approach. Our data suggest that a small region of 42 amino acids (residues 191-233) in Slob is essential for Slob to interact with the dSlo channel. PMID- 14529711 TI - Interactions between Src family protein tyrosine kinases and PSD-95. AB - Five members of the Src family of non-receptor protein tyrosine kinases--Lck, Lyn, Fyn, Src, and Yes--are known to be expressed in the central nervous system. Src and Fyn have been shown to play important roles in synaptic transmission and plasticity at excitatory synapses. Here we investigate the subcellular distribution and potential binding partners of Src family protein tyrosine kinases in brain, focusing on the lesser studied kinases Lck, Lyn, and Yes. We find that Lck, Lyn, and Yes are localized to the postsynaptic density (PSD), the primary structural component of excitatory synapses. Lyn and Yes, as well as Src, but not Lck physically associate with the prominent PSD scaffolding protein PSD 95 in co-immunoprecipitation experiments. Further, we demonstrate that PSD-95 GST fusion proteins bind directly to purified recombinant Lyn, Src, and Yes in vitro. In addition, we show that PSD-95 is unique among PSD-95 family members in that the other members, PSD-93, SAP97, and SAP102, do not physically associate with Lyn, Src, or Yes. Together our results suggest that PSD-95 may be important for localizing and/or regulating multiple Src protein tyrosine kinases at the NMDA receptor multiprotein complex. PMID- 14529712 TI - Differential binding of the AP-2 adaptor complex and PSD-95 to the C-terminus of the NMDA receptor subunit NR2B regulates surface expression. AB - NMDA receptor expression on the plasma membrane and at synaptic sites is tightly regulated. We have recently shown that the NMDA receptor subunit NR2B has an endocytic motif contained within its C-terminus. We now identify this motif as a consensus tyrosine-based motif (YEKL) and demonstrate that this sequence binds directly to the medium chain of the AP-2 adaptor, a protein complex that links internalized proteins to clathrin. Although the AP-2 binding site on NR2B is adjacent to the PSD-95 binding site, it is distinct, as mutation of tyrosine 1472 of the endocytic motif disrupts AP-2 binding but not binding to PSD-95. Internalization assays reveal that like PSD-95, both SAP97 and PSD-93 inhibit NR2B-mediated endocytosis. Furthermore, we find that co-expression of a PSD-95 mutant that is unable to cluster NMDA receptors also inhibits NR2B-mediated endocytosis. Together, these data demonstrate that AP-2 and PSD-95 bind to unique sites on the C-terminus of NR2B and have antagonistic functional consequences that are independent of the ability of the PSD-95 to cluster receptors on the plasma membrane. PMID- 14529713 TI - Disruption of the NMDA receptor-PSD-95 interaction in hippocampal neurons with no obvious physiological short-term effect. AB - PSD-95 binds to and co-localizes with NMDA receptors at postsynaptic sites. Their co-expression in COS7 cells induces the formation of aggregates containing both proteins. These findings have lead to the hypothesis that PSD-95 helps to cluster NMDA receptors at postsynaptic sites. In addition, PSD-95 binds various regulatory proteins including Src, Pyk2, SynGAP, and nNOS and may recruit signaling proteins to NMDA receptors. We tested whether synaptic transmission or plasticity was affected by acute dissociation of the PSD-95-NMDA receptor interaction with various peptides that bound to the first two PDZ domains of PSD 95 and its homologs and with antibodies directed against the very C-terminus of the NR2A and NR2B subunits of the NMDA receptor. Membrane-impermeable peptides injected via whole cell patch electrodes distributed within minutes throughout dendritic branches into spines in acute hippocampal slices and membrane-permeable peptides containing 11 arginine residues effectively accumulated in neurites in slices and primary hippocampal cultures. Neither peptides nor antibodies showed any effect on basal synaptic transmission or induction of long-term potentiation (LTP) in hippocampal slices. Pharmacologically isolated NMDA receptor activity was also not affected. However, the membrane-permeable peptide disrupted the NMDA receptor-PSD-95 interaction in slices as tested by immunoprecipitation and subsequent immunoblotting. These findings suggest that acute dissociation of PSD 95 and its homologs from the NMDA receptor and likely from other protein complexes does not result in any easily detectable physiological effects in hippocampal slices. However, we cannot exclude a role of PSD-95 in early events that lead to clustering of NMDA receptors or of other proteins including stargazin and AMPA receptors at postsynaptic sites nor do these experiments address the possibility of long-term changes in the slices. In fact, incubation of primary hippocampal cultures with the membrane-permeable peptide lead to a moderate decrease in the number of dendritic clusters of PSD-95 and NMDA receptors and their colocalization by 20-30%, suggesting some role of PSD-95 and its homologs in NMDA receptor clustering. PMID- 14529714 TI - Coordinated PKA and PKC phosphorylation suppresses RXR-mediated ER retention and regulates the surface delivery of NMDA receptors. AB - Endoplasmic reticulum (ER) retention mediated by the RXR (Arg-X-Arg) motif is an important quality control mechanism used by G-protein coupled receptors and ion channels, including N-methyl-D-aspartate (NMDA) receptors, to ensure the proper assembly and trafficking of multimeric complexes. During assembly, RXR motifs are masked by intersubunit interactions thereby allowing ER release. Here, we find that PKA and PKC phosphorylation sites flanking the RXR motif of the NMDA receptor NR1 subunit suppress ER retention and regulate receptor forward trafficking. These sites are differentially phosphorylated during the trafficking of NR1 subunits in vivo, and phosphorylation at these sites occurs in early secretory compartments. In addition, residues near the RXR motif not involved in phosphorylation are also required for ER retention. These results indicate that ER retention of NMDA receptors is tightly regulated, and suggest that coordinated phosphorylation by PKA and PKC mediates release of receptors from the ER for subsequent traffic to synapses. Phosphorylation-induced ER export of RXR containing channels and receptors may serve as a novel quality control mechanism for creating a readily releasable pool of receptors sensitive to the activation of intracellular signaling pathways. PMID- 14529715 TI - Mutations in the ligand-binding and pore domains control exit of glutamate receptors from the endoplasmic reticulum in C. elegans. AB - The abundance of ion channels and neurotransmitter receptors in the plasma membrane is limited by the efficiency of protein folding and subunit assembly in the endoplasmic reticulum (ER). The ER has a quality-control system for monitoring nascent proteins, which prevents incompletely folded and assembled proteins from being transported from the ER. Chaperone proteins identify unfolded and misassembled proteins in the ER via retention motifs that are normally buried at intersubunit contacts or via carbohydrate residues that are attached to misfolded domains. Here, we examined the trafficking of a C. elegans non-NMDA glutamate receptor (GLR-1). We show that mutations in the pore domain (predicted to block ion permeation) and mutations in the ligand-binding domain (predicted to block glutamate binding) both caused a dramatic reduction in the synaptic abundance of GLR-1 and increased retention of GLR-1 in the ER. These results suggest that the structural integrity of the ligand-binding site and the pore domain of GLR-1 are monitored in the ER during the process of quality control. PMID- 14529716 TI - Evidence for SNARE zippering during Ca2+-triggered exocytosis in PC12 cells. AB - SNAREs (soluble NSF attachment protein receptors) are membrane proteins that catalyze membrane fusion. SNAREs are defined by a characteristic 70 residue sequence called the SNARE motif. During synaptic vesicle fusion, the single SNARE motif of the synaptic vesicle SNARE protein synaptobrevin/VAMP associates into a four-helical bundle with SNARE motifs from the plasma membrane SNARE proteins syntaxin 1 and SNAP-25. The four SNARE motifs (one each from synaptobrevin and syntaxin, and two from SNAP-25) assume a parallel orientation in the complex, suggesting that formation of the complex initiates fusion by forcing the membranes containing the SNAREs into close proximity. It has been proposed that SNARE complexes assemble in an N- to C-terminal progression, a process referred to as zippering, but little direct evidence for zippering exists. Furthermore, the SM protein Munc18-1, which binds to syntaxin 1 and is essential for synaptic fusion, is thought to prepare SNAREs for complex formation by an unknown mechanism, possibly by nucleating zippering. We now show that fragments containing the N- and C-terminal regions of the SNARE motif from syntaxin 1A bind SNAP-25 similarly. However, in permeabilized PC12 cells which are used as a biochemical model system to study synaptic fusion, only fragments containing the N-terminal region are powerful inhibitors of fusion. Furthermore, mutations in the N-terminal part of the Syntaxin SNARE motif have only a moderate effect on SNAP-25 binding but abolish the inhibitory activity of the SNARE motif. Finally, larger fragments of syntaxin 1A that strongly bind to Munc18-1 but do not readily assemble into SNARE complexes had no effect on exocytosis in permeabilized PC12 cells. Together these results suggest that Munc18-1 acts before SNARE complex assembly, and is no longer required at the stage of fusion assayed in permeabilized PC12 cells. The selective effect of the N-terminal half of the syntaxin 1A SNARE motif on PC12 cell exocytosis shows that the SNARE motif is functionally polarized, and supports the notion that SNARE complexes assemble in an N- to C-terminal zippering reaction during fusion without a stable, partially assembled intermediate. PMID- 14529717 TI - A putative role for intramolecular regulatory mechanisms in the adaptor function of amphiphysin in endocytosis. AB - Amphiphysin 1 is a brain-specific protein enriched at the synapse and a major binding partner of several components of the clathrin-mediated endocytic machinery (Proc Natl Acad Sci USA 93 (1996) 331). It interacts with clathrin-coat proteins, dynamin, and membranes (Nat Cell Biol 1 (1999) 33; JBC). A role of amphiphysin in synaptic vesicle recycling is supported by both acute and chronic perturbation studies (Science 276 (1997) 259; Neuron 33 (2002) 789). Here we show that amphiphysin directly stimulates clathrin recruitment onto liposomes in an in vitro assay. Amphiphysin-dependent clathrin-coat recruitment is enhanced by the interaction of amphiphysin with dynamin. We also show that the amphiphysin SH3 domain binds full-length amphiphysin, likely via an internal poly-proline region, and that clathrin recruitment onto liposomes by amphiphysin is enhanced in the presence of the isolated amphiphysin SH3 domain. Expression of a mutant amphiphysin harboring two amino acid substitutions in the SH3 domain, and therefore unable to bind proline-containing motifs, induces an accumulation of large intracellular aggregates including amphiphysin, clathrin, AP-2, and other endocytic proteins, as well as a concomitant block of transferrin endocytosis. Thus, putative intramolecular interactions between the amphiphysin COOH-terminal SH3 domain and its internal poly-proline region may regulate clathrin recruitment onto membranes. PMID- 14529718 TI - Imaging single synaptic vesicles undergoing repeated fusion events: kissing, running, and kissing again. AB - At synapses of the mammalian central nervous system, release of neurotransmitter occurs at rates transiently as high as 100 Hz, putting extreme demands on nerve terminals with only tens of functional vesicles at their disposal. Thus, the presynaptic vesicle cycle is particularly critical to maintain neurotransmission. To understand vesicle cycling at the most fundamental level, we studied single vesicles undergoing exo/endocytosis and tracked the fate of newly retrieved vesicles. This was accomplished by minimally stimulating boutons in the presence of the membrane-fluorescent styryl dye FM1-43, then selecting for terminals that contained only one dye-filled vesicle. We then observed the kinetics of dye release during single action potential stimulation. We found that most vesicles lost only a portion of their total dye during a single fusion event, but were able to fuse again soon thereafter. We interpret this as direct evidence of "kiss and-run" followed by rapid reuse. Other interpretations such as "partial loading" and "endosomal splitting" were largely excluded on the basis of multiple lines of evidence. Our data placed an upper bound of <1.4 s on the lifetime of the kiss and-run fusion event, based on the assumption that aqueous departitioning is rate limiting. The repeated use of individual vesicles held over a range of stimulus frequencies up to 30 Hz and was associated with neurotransmitter release. A small percentage of fusion events did release a whole vesicle's worth of dye in one action potential, consistent with a classical picture of exocytosis as fusion followed by complete collapse or at least very slow retrieval. PMID- 14529719 TI - CaVbeta subunit-mediated up-regulation of CaV2.2 currents triggered by D2 dopamine receptor activation. AB - Voltage-dependent Ca(2+) channels (VDCCs) are subject to modulation by a number of pathways, including membrane-delimited inhibition by heterotrimeric G-proteins and modulation through phosphorylation by diverse kinases. Here we report that in the Xenopus oocyte expression system Ca(V)2.2 channels undergo a sustained, linear and irreversible run-up lasting up to 30 min, which is potentiated during G-protein-mediated inhibition by activation of co-expressed G-protein coupled receptors (GPCRs). This up-regulation is not a result of receptor desensitization, but is associated with a hyperpolarization of the voltage for activation and depends on the presence of accessory subunits such that beta subunits promote, and alpha2delta subunits oppose the current increase. We have investigated the involvement of G-proteins and found that over-expression of Galpha(o) subunits or Galpha-transducin reduced the amount of agonist-mediated up regulation. However, we have found no evidence for the involvement of any second messenger pathways in the increase of current run-up in the presence of a GPCR agonist. Taken together, our data suggest that the effect reported herein involves an enhancement of the GTPase activity of endogenous Galpha subunits, which is triggered by GPCR activation and mediated by accessory Ca(V)beta subunits. It may involve an increased association of Ca(V)beta subunits with alpha1 subunits in the plasma membrane or trafficking of channels to the plasma membrane. PMID- 14529720 TI - Parallel kinase cascades are involved in the induction of LTP at hippocampal CA1 synapses. AB - To identify the enzymes involved in the induction of N-methyl-D-aspartate (NMDA) receptor-dependent long-term potentiation (LTP) at CA1 synapses of two-week-old rats we have tested various kinase inhibitors. Surprisingly, given the large body of evidence supporting a role for calcium/calmodulin-dependent protein kinase II (CaMKII) in LTP, inhibition of this enzyme did not affect the induction of LTP at this age. Similarly inhibition of protein kinase A (PKA) or protein kinase C (PKC) was also without effect. However, inhibition of CaMKII together with inhibition of either PKA or PKC fully blocked the induction of LTP. These experiments reveal, unexpectedly, the existence of two parallel kinase pathways, one involving CaMKII and the other PKA and PKC, either of which can fully support the induction of LTP, at this stage of development. PMID- 14529721 TI - The PDZ domains of mLin-10 regulate its trans-Golgi network targeting and the surface expression of AMPA receptors. AB - Dynamic regulation of synaptic AMPA receptor localization underlies certain forms of synaptic plasticity and researchers are just beginning to identify molecules that may play a role in the synaptic delivery of glutamate receptors. One candidate is mLin-10, the mammalian homolog of the C. elegans receptor targeting protein LIN-10. Here, we investigated the role of mLin-10 in glutamate receptor trafficking. Cellular localization studies, in both whole brain and cultured neurons, revealed that mLin-10 is enriched in the trans-Golgi network and present in dendrites and spines--regions where protein sorting and synaptic delivery are known to occur. The specific localization of mLin-10 in Golgi is disrupted by a point mutation in an mLin-10 PDZ domain, indicating that a PDZ domain mediates this localization. Interactions between mLin-10 and glutamate receptors in both intracellular and synaptic membrane fractions were detected through biochemical assays. GST-pull down and co-immunoprecipitation experiments in heterologous cells delineated the protein domains required for interaction. These results demonstrated that glutamate receptors interact directly with mLin-10 through a PDZ domain-mediated mechanism. A PDZ point mutation enhances surface delivery of exogenous glutamate receptors in transfected neurons, suggesting that mLin-10 may regulate AMPA receptor trafficking in vivo. PMID- 14529722 TI - Interaction of transmembrane AMPA receptor regulatory proteins with multiple membrane associated guanylate kinases. AB - Surface expression of AMPA type glutamate receptors requires stargazin or a related transmembrane AMPA receptor regulatory protein (TARP). Furthermore, interaction of the cytosolic tail of TARPs with PDZ domains of PSD-95 targets AMPA receptors to postsynaptic densities. Here, we screened for additional proteins that might interact with the cytosolic domain of TARPs. Screening a rat brain cDNA library with the yeast two-hybrid system yielded six PDZ proteins that can bind tail of TARPs. These PDZ proteins include the four neuronal membrane associated guanylate kinases, PSD-95/SAP-90, PSD-93/Chapsyn-110, SAP-97/hDLG and SAP-102; the multi-PDZ protein, MUPP1; and the mitochondrial PDZ protein, OMP-25. Although all of these proteins can bind to TARPs in vitro, only two of these, PSD 95 and PSD-93 associate with TARPs in brain. We also found that all three PDZ domains from PSD-95 associate with the TARP C-termini with similar affinities. This work identifies biochemical promiscuity for interaction of the TARP C termini with PDZ domains in vitro, but also shows that only specific synaptic PDZ proteins associate with TARPs in brain. PMID- 14529723 TI - SNAP-25 Ser187 does not mediate phorbol ester enhancement of hippocampal synaptic transmission. AB - Phorbol esters, activators of protein kinase C (PKC), have been shown to enhance synaptic transmission. One potential downstream target of PKC in the presynaptic terminal is the soluble N-ethylmaleimide sensitive factor (NSF) attachment protein receptor (SNARE) SNAP-25, which has a PKC phosphorylation site in its C terminal coil centered at serine 187 (S187/Ser187). We examined the role of S187 in hippocampal synaptic transmission. After proteolytic cleavage of native SNAP 25 by botulinum neurotoxin E (BoNT/E), synaptic transmission was restored in a subset of transfected CA3 pyramidal cells with a toxin-resistant form of SNAP-25 containing unaltered S187 (Swt), S187 mutated to alanine (SA) or S187 mutated to glutamate (SE). We observed that phorbol-12,13-diacetate (PDAc, 10 microM) induced potentiation of neurotransmission to a similar degree for both Swt and SA (2.4-fold and 3.1-fold increase, respectively). Furthermore, basal levels of transmission mediated by SE were reduced relative to that of Swt (failure rates of 72% and 41%, respectively). Together, these data suggest that phosphorylation of SNAP-25 S187 does not mediate the observed enhancement of neurotransmission by phorbol esters at hippocampal synapses. PMID- 14529724 TI - ERKs regulate PKC-dependent synaptic depression and declustering of glutamate receptors in cerebellar Purkinje cells. AB - Phorbol esters, such as tetradecanoylphorbol 13-acetate (TPA), have been used extensively in studies of cerebellar long-term depression (LTD), based on the hypothesis that activated protein kinase C (PKC) directly mediates alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor phosphorylation. Here, we show that TPA-induced depression of synaptic transmission between granule cells and Purkinje cells in culture is mediated through activation of the MEK1/2 ERK1/2 pathway. Phosphorylation of ERK1/2 induced by TPA and co-application of high potassium and glutamate was greatly attenuated by preincubating Purkinje cells with the MEK1/2 (MAPK ERK kinase 1/2) inhibitor PD98059. TPA-induced depression of synaptic transmission between granule cells and Purkinje cells was attenuated by PD98059. The MEK1/2 inhibitor also suppressed declustering of the ionotropic glutamate receptor subunit 2/3 (GluR2/3) induced by TPA and co application of high potassium and glutamate, even though phosphorylation of Ser880 of GluR2/3 was not inhibited significantly in the presence of PD98059. These results suggest that ERK1/2 plays an essential role in TPA-induced depression via regulation of GluR2/3 declustering at the synapse. PMID- 14529725 TI - Phosphorylation influences neurosteroid modulation of synaptic GABAA receptors in rat CA1 and dentate gyrus neurones. AB - The neurosteroid 5beta-pregnan-3alpha-ol-20-one (5beta3alpha) is a potent, endogenous, positive allosteric modulator of the GABA(A) receptor. Relatively low concentrations of 5beta3alpha (10-100 nM), thought to occur physiologically, caused a concentration-dependent slowing of the decay of GABA-mediated miniature inhibitory postsynaptic currents (mIPSCs) recorded from hippocampal CA1 pyramidal neurones. However, much greater concentrations of this neurosteroid (> or =300 nM) were required to similarly influence dentate granule cell mIPSCs. By contrast, the allosteric modulators pentobarbitone and flunitrazepam were equi effective in prolonging mIPSCs in both neuronal types. Hence, the neurosteroid selectively differentiates between the synaptic GABA(A) receptors of these hippocampal neurones. Inhibition of either protein kinase A, or C, greatly reduced the sensitivity of CA1 synaptic GABA(A) receptors to 5beta3alpha, but not pentobarbitone, whereas stimulation of PKC had no effect on steroid sensitivity. However, in dentate gyrus granule cells, activation of PKC made mIPSCs sensitive to a previously ineffective concentration of 5beta3alpha. Collectively, these results suggest that the GABA-modulatory effects of physiological levels of the neurosteroid will not be uniformly experienced throughout the central nervous system, or even within the same brain region such as the hippocampus, but will be neurone-specific and will be dependent on the phosphorylation status of the GABA(A) receptor, or associated proteins. PMID- 14529726 TI - Pharmacological characterization of the presynaptic activity of Tityus serrulatus venom in the rat anococcygeus muscle. AB - Scorpion venoms are known to cause peripheral nerve stimulation with enhanced autonomic responses. This study, therefore, examined the effects of Tityus serrulatus venom (TSV) on adrenergic, cholinergic and nitrergic nerve fibers using the rat anococcygeus muscle. The contractile effects of TSV (1 microg/ml) and electrical field stimulation were markedly reduced by phentolamine (5 microM), prazosin (0.1 microM), guanethidine (30 microM) and tetrodotoxin (TTX, 1 microM), whereas imipramine (3 microM) enhanced these responses. The responses to tyramine (10 microM) were partially reduced by guanethidine and completely blocked by phentolamine, prazosin and imipramine. Atropine (1 microM) fully prevented carbachol (CCh, 30 microM)-induced contractions without affecting those mediated by TSV. Neostigmine significantly potentiated TSV-and ACh-evoked contractions, whereas hexamethonium had no effect. The relaxant responses induced by EFS and TSV (3 microg/ml) were completely blocked by L-NAME (100 microM), ODQ (1 microM) or TTX (1 microM). Addition of L-arginine (1 mM) reversed the effect of L-NAME. Thus, the motor and inhibitory responses of TSV in the rat anococcygeus muscle are mediated by prejunctional mechanisms dependent on Na(+) channel activation, causing the stimulation of NA and NO release from adrenergic and nitrergic nerve fibers, respectively. PMID- 14529727 TI - Intramuscular injection of 125I-botulinum neurotoxin-complex versus 125I botulinum-free neurotoxin: time course of tissue distribution. AB - The diffusion from the site of intramuscular injection of 900 kDa botulinum neurotoxin-hemagglutinin complex (BoNT/A-complex) and 150 kDa free-botulinum neurotoxin (free-BoNT/A) was compared. Radioiodinated compounds were injected into the gastrocnemius muscle of rats (70Units (U) 125I-BoNT/A-complex, 67 or 344 U free-125I-BoNT/A, or free-125I-iodide) and the eyelids of rabbits (24 U 125I BoNT/A-complex or 108 U free-125I-BoNT/A), and measured in various tissues at different time points. There were no detectable systemic effects or generalized botulinum neurotoxin toxicity in either rats or rabbits, indicating that most of the toxin, whether as 125I-BoNT/A-complex or free-125I-BoNT/A, remained at the injection site. In rats, 125I-BoNT/A-complex and free-125I-BoNT/A diffused in a pattern that was grossly similar. Almost no radioactivity was recovered from the brain. Radioactivity recovered from distant tissues (thyroid, skin, and contralateral muscle) was primarily attributable to either low molecular weight 125I-containing peptides or 125I-iodide. After injection into rabbit eyelids, neither 125I-BoNT/A-complex nor free-125I-BoNT/A spread to distant structures, including the eye. The results indicate that most of the neurotoxin does not diffuse from the injection site, whether in free or complexed form, and this may reduce the potential for systemic effects. PMID- 14529728 TI - Influence of sphingomyelin and TNF-alpha release on lethality and local inflammatory reaction induced by Loxosceles gaucho spider venom in mice. AB - It is well known that Loxosceles venom induces local dermonecrosis in rabbits, guinea pigs and humans but not in mice, although, depending on the dose, Loxosceles venom can be lethal to mice. In this work we demonstrate that mice injected intradermally in the dorsal area of the back can survive a lethal dose of Loxosceles gaucho venom and also develop an inflammatory reaction (with infiltration of leukocytes shown by histological analysis) at the local injection site when the venom is co-administered with sphingomyelin. It was observed that more venom was retained for a longer period of time at the local injection site when venom was co-administered with sphingomyelin. The presence of exogenous sphingomyelin did not influence significantly the release of TNF-alpha induced by L. gaucho venom. These results suggest that the action of venom on sphingomyelin, producing ceramide phosphate, causes the development of an inflammatory reaction, which in turn traps the venom in the local area for a long period of time and does not allow it to disperse systemically in a dose sufficient to cause death. Our findings also indicate that the size and availability of the local sphingomyelin pool may be important in determining the outcome of Loxosceles envenoming in different mammalian species. PMID- 14529729 TI - Characterization of nerve growth factors (NGFs) from snake venoms by use of a novel, quantitative bioassay utilizing pheochromocytoma (PC12) cells overexpressing human trkA receptors. AB - Snake venoms are a very abundant source of nerve growth factors (NGF). NGFs of Elapidae showing 65% sequence homology with mouse or human NGF, while the Viperidae NGF shows N-glycosylation (Asn-21) typical of these mammalian NGFs. Snake NGF-induced neurite outgrowth (neurotropic activity) was measured in the past by using PC12 cell or dorsal root ganglion bioassays. The present study was aimed at comparing, by dose-response experiments, the neurotropic activity of cobra and vipera versus mammalian NGFs, by using a novel bioassay involving PC12 cells genetically engineered to overexpress NGF-trkA receptors of human origin. These cells respond to NGF by differentiation (morphologically expressed as neurite outgrowth) by a process mediated by NGF-trkA receptors. This process was evaluated by two different criteria: (1) elongation of neurites (E), and (2) Percentage of responsive cells (PRC) determined by digital acquisition of data and computer analysis. We found that snake venom NGFs were less potent than mouse NGF, and that cobra NGF was more potent than vipera NGF. These data indicate the following order of NGF activity towards recombinant human trkA receptors: recombinant human NGF>mouse NGF>cobra NGF>vipera NGF. The neurotropic efficacy of these NGFs was found to be similar, reaching 80-90% of maximal activity obtained with all NGF forms. Interestingly, cobra (but not vipera) NGF demonstrated prolonged neurotropic activity compared with mouse NGF. The results of the present study indicate that cobra and vipera venom NGFs represent natural agonists of human trkA-receptor of a lower potency, but of similar efficacy, compared with mammalian NGFs. These compounds are important pharmacological tools to characterize the trkA receptor structure-function relationship, and to develop novel neurotropic drugs. PMID- 14529730 TI - Primary structure of echotoxin 2, an actinoporin-like hemolytic toxin from the salivary gland of the marine gastropod Monoplex echo. AB - Echotoxins are 25 kDa proteins with both hemolytic and lethal activities, previously purified from the salivary gland of the marine gastropod Monoplex echo. In this study, a cDNA encoding echotoxin 2 was cloned by RT-PCR, 3'-RACE and 5'-RACE, based on its partial amino acid sequence. The full-length echotoxin 2 cDNA (1000 bp) obtained contains an open reading frame (825 bp) coding for a precursor protein of 274 amino acid residues. Mature echotoxin 2 composed of 226 amino acid residues is assumed to be produced by post-translational removal of N terminal 23 residues (predicted as a signal peptide) and C-terminal 25 residues from the precursor protein. Very interestingly, a homology search revealed that echotoxin 2 is analogous to actinoporins, 20 kDa pore-forming hemolysins reported from various sea anemones. In addition to the similarities in biological activity, molecular size and basicity between echotoxin 2 and actinoporins, two prominent structural features, an N-terminal amphiphilic alpha-helix and an aromatic patch comprising Trp and Tyr residues, both of which are important for the pore-forming activity of actinoporins, are also recognized in echotoxin 2. However, echotoxin 2 is distinguishable from actinoporins in having Cys residues and lacking an RGD motif. PMID- 14529731 TI - Characterisation of local inflammatory response induced by Thalassophryne nattereri fish venom in a mouse model of tissue injury. AB - The Thalassophryne nattereri fish venom induces a severe burning pain, oedema, and necrosis observed both clinically and experimentally. The present study was carried out in order to describe the pattern of local acute inflammatory response after T. nattereri venom injection. Our findings show that the edematogenic response induced by T. nattereri venom in footpad of mice was dose- and time dependent, and remained significantly elevated over 48 h after injection. Analysis of footpad homogenates were tested for the presence of TNF-alpha, IL 1beta and IL-6, and demonstrated augmented levels of these cytokines. Our results showed that the injection of venom developed an inadequate cellular inflammatory response evidenced by poor infiltration of mononuclear cells, preceded by decreased number of these cells in peripheral blood. In contrast, we observed an early intense recruitment of neutrophil to peritoneal cavity, accompanied by a significant decrease in the number of mononuclear cells. A drastic increase in the total amount of cells, mainly in neutrophils, followed by mononuclear cell recruitment was observed 24 h. In addition, we also demonstrated that T. nattereri venom affects the viability of mononuclear cells (J774A1) in culture. We conclude that the scarcity of inflammatory cellular influx into local lesions (intraplantar) induced by T. nattereri venom could be a consequence of an impaired blood flow in venules at injured tissue and cytotoxic effect of the venom on inflammatory cells can contribute to this impairment. PMID- 14529732 TI - Effects of Thalassophryne nattereri fish venom in isolated perfused rat kidney. AB - Thalassophryne nattereri, popularly known as Niquim, is a venomous fish responsible for many accidents in fishermen in the Northeast of Brazil. The effects of T. nattereri venom on renal physiology has not been tested. Isolated kidneys from Wistar rats of 240-280 g weight were perfused with Krebs-Henseleit solution containing 6g% of previously dialyzed bovine serum albumin. The effects of Niquim venom were studied on the perfusion pressure (PP), renal vascular resistance (RVR), urinary flow (UF), glomerular filtration rate (GFR), percent of sodium tubular transport (%TNa(+)), percent of potassium tubular transport (%TK(+)) and percent of chloride tubular transport (%TCl(-)). The venom of T. nattereri (0.3, 1.0, and 3.0 microg/ml) was always added to the system 30 minutes after the beginning of each experiment (n=6). All experiments were preceded by 30 minutes internal control period and an external control group, where kidneys were perfused with only Krebs-Henseleit solution. All three doses tested promoted increases in PP and RVR. The first two doses also increased GFR and UF. The higher dose promoted decreases in GFR, UF, %TNa(+), %TK(+), %TCl(-). In the treated groups we observed hyalin casts inside all tubules and proteinaceous material in the urinary space. We conclude that the effects resulted from niquim venom agents that promoted a direct effect in kidney cells causing the release of vasoactive factors. PMID- 14529733 TI - Pharmacological characterization of mouse hind paw oedema induced by Bothrops insularis (jararaca ilhoa) snake venom. AB - Bothrops snake venoms produce marked local effects, including oedema, haemorrhage and necrosis. The ability of Bothrops insularis venom to induce oedema in mice was investigated. Venom was injected into hind paws and the change in volume over time was measured by plethysmometry. B. insularis venom (0.01-2.5 microg/paw) induced paw oedema which, at high doses (>/=0.5 microg/paw), was accompanied by haemorrhage. The peak oedematogenic response occurred 3 h after venom injection with all doses and decreased gradually thereafter, but was still elevated with high doses after 24 h. Pretreating the mice with cyproheptadine (histamine H(1) and serotonin 5-HT(2) receptor antagonist), mepyramine (histamine H(1) receptor antagonist), L-NAME (inhibitor of nitric oxide synthase), indomethacin and rofecoxib (inhibitors of cyclooxygenases), and dexamethasone (indirect inhibitor of PLA(2)) significantly attenuated venom-induced oedema, whereas methysergide, a serotonin 5-HT(1)/5-HT(2) receptor antagonist, had no effect. The administration of antivenom 30 min before or immediately after venom injection also significantly inhibited venom-induced oedema. These results show that B. insularis venom causes oedema in the mouse hind paw and that this response is mediated by histamine, nitric oxide, and arachidonic acid metabolites formed by cyclooxygenases 1 and 2. The neutralization by commercial antivenom indicates that the venom components responsible for oedema are recognized by the antivenom and share immunological identity with their counterparts in the venoms of mainland Bothrops species. PMID- 14529734 TI - Purification and structural characterization of lectins from the cnidarian Bunodeopsis antilliensis [corrected]. AB - Purification and characterization of two different lectins from the Mexican anemone Bunodeopsis antilliensis are reported. These two lectins named Bunodeopsis antilliensis agglutinin-A (BAA-A) and -B (BAA-B) presented the following characteristics: BAA-A was resolved as a component, with haemagglutinating activity for human blood type A (N-acetylgalactosamine galactose-fucose), with a molecular weight of 28,900 obtained by means of mass spectrometry, showed an isoelectric point of 5.04 with a higher carbohydrate specificity for N-acetylgalactosamine (GalNAc). The analysis of the N-terminal revealed it is related to phosphoesterase and GTP binding protein. BAA-B mainly active with human blood type B (galactose-galactose-fucose) was resolved into three fractions (BAA-B1-3). Their molecular weight were: BAA-B(1) 39,350, BAA B(2) 28,300 and BAA-B(3) 17,550. The estimated isoelectric points were 8.05, 4.66 and 6.60, respectively. Only fraction 3 exhibited haemagglutinating activity with a higher carbohydrate specificity for galactose and mannose. The analysis of the N-terminal pointed out it is related with phospholipase A(2). We suggest these lectins could be related to a feeding strategy[corrected]. PMID- 14529735 TI - Simultaneous production of homoanatoxin-a, anatoxin-a, and a new non-toxic 4 hydroxyhomoanatoxin-a by the cyanobacterium Raphidiopsis mediterranea Skuja. AB - A neurotoxin, homoanatoxin-a, was identified from a toxic strain of the cyanobacterium Raphidiopsis mediterranea Skuja (strain LBRI 48) isolated from Lake Biwa, Japan, as the major toxin component (0.57% of dry cell-weight). This cyanobacterium produced anatoxin-a and a new homoanatoxin-a derivative as minor components (0.04 and 0.06%, respectively). The structure of a new compound was assigned based on the spectral data as 4-hydroxyhomoanatoxin-a, which was not toxic to mice up to 2.0 mg/kg by intraperitoneal injection. The isolation of minor components was accomplished by improved extraction and separation procedures: (1) extraction with methanol-water (4:1) from dried cells, (2) adsorption of aqueous residue on ODS column (or cartridge) after evaporation of methanol, (3) cleaning up of the column by successive elution with water and 50% methanol/water, (4) elution of a toxic fraction by 20% methanol/water containing 0.1% TFA and (5) HPLC (ODS) purification with methanol/water containing 0.05% TFA. The procedures were effective in removing impurities and concentrating alkaloidal neurotoxins. It should be noted that this is the first report demonstrating the simultaneous production of anatoxin-a and homoanatoxin-a by the same strain of cyanobacterium. PMID- 14529736 TI - Purification and cloning of cysteine-rich proteins from Trimeresurus jerdonii and Naja atra venoms. AB - Three 26 kDa proteins, named as TJ-CRVP, NA-CRVP1 and NA-CRVP2, were isolated from the venoms of Trimeresurus jerdonii and Naja atra, respectively. The N terminal sequences of TJ-CRVP and NA-CRVPs were determined. These components were devoid of the enzymatic activities tested, such as phospholipase A(2), arginine esterase, proteolysis, L-amino acid oxidase, 5'nucleotidase, acetylcholinesterase. Furthermore, these three components did not have the following biological activities: coagulant and anticoagulant activities, lethal activity, myotoxicity, hemorrhagic activity, platelet aggregation and platelet aggregation-inhibiting activities. These proteins are named as cysteine-rich venom protein (CRVP) because their sequences showed high level of similarity with mammalian cysteine-rich secretory protein (CRISP) family. Recently, some CRISP like proteins were also isolated from several different snake venoms, including Agkistrodon blomhoffi, Trimeresurus flavoviridis, Lanticauda semifascita and king cobra. We presumed that CRVP might be a common component in snake venoms. Of particular interest, phylogenetic analysis and sequence alignment showed that NA CRVP1 and ophanin, both from elapid snakes, share higher similarity with CRVPs from Viperidae snakes. PMID- 14529738 TI - Biological activity of 8,11-dideoxytetrodotoxin: lethality to mice and the inhibitory activity to cytotoxicity of ouabain and veratridine in mouse neuroblastoma cells, Neuro-2a. AB - Contribution of the C-8 hydroxyl group of tetrodotoxin to its sodium channel blocking activity has never been clearly evaluated. Isobe et al. recently synthesized 8,11-dideoxytetrodotoxin, the first 8-deoxy analog of tetrodotoxin. In this study, the biological activity of this compound was investigated to compare with that of 11-deoxytetrodotoxin. Intraperitoneal injection of 8,11 dideoxytetrodotoxin at the level of 700 microg/kg did not kill a mouse (n=2), indicating that the lethal dose of this compound was more than 70 and 10 folds larger than LD(50) of tetrodotoxin and 11-deoxytetrodotoxin, respectively. The inhibitory activity of 8,11-dideoxytetrodotoxin to cytotoxicity of ouabain and veratridine in mouse neuroblastoma cells (Neuro-2a) was also examined. The ED(50) for 8,11-dideoxytetrodotoxin was estimated to be 9.3+/-3.3 microM (n=3), approximately 2000 and 34 folds larger than those of tetrodotoxin (4.6+/-0.70 nM, n=3) and 11-deoxytetrodotoxin (270+/-74 nM, n=4), respectively. These data suggest that the C-8 hydroxyl group of tetrodotoxin is also important for its activity, as well as all the other hydroxyl groups. PMID- 14529739 TI - The equine antitoxins supply system for biological poisons in Japan. AB - Recently, the equine antitoxin supply in Japan has sharply decreased; then it is apparent that a stable supply produced solely by private industry cannot be relied upon. The Ministry of Health, Labor and Welfare (MHLW), therefore, purchases vaccines and equine antitoxins from manufacturers who could not otherwise independently provide an adequate antitoxin supply to hospitals. This supply system is called the 'Kokuyu vaccine system.' Under this system, MHLW purchases, stores and distributes vaccines and antitoxins to hospitals. This system has worked efficiently and effectively so far and may be a good model for establishing a stable antitoxin supply system in other countries. PMID- 14529737 TI - New insight on scorpion divergence inferred from comparative analysis of toxin structure, pharmacology and distribution. AB - The divergence of Buthidae, the most abundant family of scorpions, has relied thus far on anatomical and morphological features, but still remains controversial. However, much information has accumulated on Buthidae long-chain scorpion toxins affecting neuronal sodium channel conductance (alpha- and beta toxins) and their pharmacology. Therefore, we constructed a toxin evolutionary tree, which together with recent data on toxin gene organization, toxin structures, and worldwide dispersion, sheds light on toxin and hence, scorpion divergence. Based on these data, we suggest that in the ancient world, the ancestral long-chain toxins affecting sodium channels developed into beta-like toxins, which most likely developed into alpha- and beta-toxins before the separation of South America from Africa. Subsequently, in the Old World, mostly excitatory and depressant toxins developed from the ancestral beta-like toxin and in the New World a new type of toxin group with beta-toxin structure but alpha toxin activity developed from the beta-toxins. Assisted by the worldwide distribution of toxins and the zoogeographical dispersion of the studied genera in Asia and Africa (Old World) and in South and North America (New World), we suggest a route of divergence for some of the Buthidae scorpions, a task that has reached a standstill when morphological and anatomical features were used. PMID- 14529740 TI - The occurrence of 11-oxotetrodotoxin, a rare tetrodotoxin analogue, in the brachycephalidae frog Brachycephalus ephippium. AB - 11-oxoTTX is an analogue 4-5 times more toxic than TTX itself, been rare even in marine animals. Two ions at m/z 320 and 336 corresponding to TTX and 11-oxoTTX (M+H(+)), respectively, were detected in the Brachycephalidae frog Brachycephalus ephippium extracts. The fragment ion pattern of 11-oxoTTX is similar to that TTX, although its possible to verify some specific fragments. PMID- 14529741 TI - Differential gene expression analysis in fish exposed to endocrine disrupting compounds. AB - This review discusses various methodologies that can be used to understand, at the gene level, the consequences to fish upon exposure to endocrine disrupting compounds (EDCs). Several approaches for measuring expression of gene transcripts are discussed, including directed approaches, such as Northern blotting and quantitative reverse transcriptase polymerase chain reaction (RT-PCR) as well as open-ended approaches, such as differential display RT-PCR, subtractive hybridizations, and gene arrays. Each of these systems has advantages and disadvantages, strengths and weaknesses. Conducting experiments with each of these methods provides important information about the molecular mechanisms that result from exposure to EDCs, information which can be used in risk assessment of polluted sites found in the environment. PMID- 14529742 TI - Molecular cloning and characterization of a hypoxia-responsive CITED3 cDNA from grass carp. AB - We have isolated a 1586-bp full-length CITED3 cDNA from grass carp which specifies for a cAMP-responsive element-binding protein/p300-interacting transactivator with glutamic acid (E)/aspartic acid (D)-rich C-terminal domain protein. The cDNA, designated as gcCITED3, has an open reading frame of 762 bp and encodes a protein of 253 amino acids with a predicted molecular mass of 28.3 kDa and pI of 6.4. Pairwise comparison showed that gcCITED3 shares high sequence identity with the CITED3 of zebrafish (94%), chicken (72%) and Xenopus (59%). Northern blot analysis indicated that gcCITED3 is most highly expressed and responsive to hypoxia in the carp kidney. Hypoxic induction was also observed in heart, albeit at a lower level. This is the first report on the isolation of a hypoxia-responsive CITED3 gene from fish. PMID- 14529743 TI - Toxicogenomic effects of marine brevetoxins in liver and brain of mouse. AB - Although the polyether brevetoxins (PbTx's) produced by Karenia brevis (the organism responsible for blooms of the Florida red tide) are known to exert their acute toxic effects through ion-channel mediated pathways in neural tissue, prior studies have also demonstrated that at least one form of the toxin (PbTx-6) is bound avidly by the aryl hydrocarbon receptor (AhR). Since AhR binding of a prototypical ligand such as dioxin is the first step in a cascade pathway producing major changes in gene expression, we reasoned that PbTx-6 might produce similar genomic-wide changes in expression. Mice were injected i.p. with sub lethal doses of PbTx's (either 1.5 or 3 mg/g body weight of PbTx-6; or 0.15 mg/g body weight of PbTx-2, a toxin not avidly bound by the AhR), and liver and brain tissues were sampled at 8, 24 and 72 h and RNA was isolated. Changes in gene specific RNA levels were assessed using commercially available mouse cDNA arrays (Incyte) containing >9600 array elements, including many elements from AhR mediated genes. Histopathology of the two organs was also assessed. We observed minor histopathological effects and a total of only 29 significant (>2.0-fold) changes in gene expression, most of which occurred in the liver, and most of which could be attributable to an 'acute phase' inflammatory response. These results argue against the hypothesis that PbTx-6 acts via a classic AhR-mediated mechanism to evoke gene expression changes. However, given the avidity with which PbTx-6 binds to the AhR, these findings have important implications for how PbTx's may act in concert with other toxicants that are sensed by the AhR. PMID- 14529745 TI - Ancient signals: peptides and the interpretation of positional information in ancestral metazoans. AB - Understanding the 'tool kit' that builds the most fundamental aspects of animal complexity requires data from the basal animals. Among the earliest diverging animal phyla are the Cnidaria which are the first in having a defined body plan including an axis, a nervous system and a tissue layer construction. Here I revise our understanding of patterning mechanism in cnidarians with special emphasis on the nature of positional signals in Hydra as perhaps the best studied model organism within this phylum. I show that (i) peptides play a major role as positional signals and in cell-cell communication; (ii) that intracellular signalling pathways in Hydra leading to activation of target genes are shared with all multicellular animals; (iii) that homeobox genes translate the positional signals; and (iv) that the signals are integrated by a complex genetic regulatory machinery that includes both novel cis regulatory elements as well as taxon specific target genes. On the basis of these results I present a model for the regulatory interactions required for axis formation in Hydra. PMID- 14529746 TI - Adipokinetic hormone inhibits the formation of energy stores and egg production in the cricket Gryllus bimaculatus. AB - Adipokinetic hormones (AKHs) mobilise lipids, carbohydrates and/or proline from insect fat body stores. In addition, AKHs inhibit lipid and protein synthesis in the fat body. In the current study, 100 pmol homologous Grybi-AKH was injected twice daily into adult female crickets, Gryllus bimaculatus, starting on the day of adult emergence. The effects of the injected AKH on the formation of energy reserves in the fat body and on egg production were measured on day 4 after adult emergence. In comparison to water-injected control animals, lipid and protein content in the fat body of the AKH-injected crickets was significantly reduced, suggesting an inhibitory effect of AKH on the formation of lipid reserves and protein stores. The content of glycogen and free carbohydrate in the fat body was significantly higher in the AKH-injected animals. The most pronounced effect of the AKH-injections was a significant reduction of ovary mass, due to the retarded maturation of the oocytes and the significantly lower number of terminal oocytes produced. It is concluded that AKH inhibits egg production indirectly by interference with the formation of energy stores in the fat body that are mobilised to fuel egg production. PMID- 14529747 TI - Expression of allatostatins in the Mediterranean field cricket, Gryllus bimaculatus de Geer (Ensifera, Gryllidae). AB - The allatostatin (AST) type A gene of the cricket Gryllus bimaculatus encodes a hormone precursor including at least 14 putative peptides with a common C terminus Y/FXFGL/Iamide. By RT-PCR we have analyzed the expression of the allatostatin precursor in various tissues of 0-21 days old adult virgin and mated females. In 3-day-old virgin females, the gene is strongly expressed in the brain (oesophageal ganglion), the suboesophageal ganglion and the caecum, but to a lower extent in other parts of the digestive tract (ileum, midgut, colon), and in various other tissues such as the fat body, ovaries and female accessory reproductive glands. In the brain and ovaries of virgin females, the AST expression is rather constant throughout adult life, whereas in brains of mated animals, expression is low until day 7, but increases sharply from day 8 onwards to reach values triple those before day 7. In ovaries of mated animals AST gene expression is also age-dependent, with high expression rates during the first 4 days after imaginable moult, a second but smaller peak from day 15 to 21, and very low values in between. In the fat body of virgin crickets allatostatin expression is high during the first 9 days after ecdysis and declines thereafter, whereas in mated animals two peak values, day 1 and day 6, are observed, and a third peak in older animals. PMID- 14529748 TI - Insect adipokinetic hormones: release and integration of flight energy metabolism. AB - Insect flight involves mobilization, transport and utilization of endogenous energy reserves at extremely high rates. Peptide adipokinetic hormones (AKHs), synthesized and stored in neuroendocrine cells, integrate flight energy metabolism. The complex multifactorial control mechanism for AKH release in the locust includes both stimulatory and inhibitory factors. The AKHs are synthesized continuously, resulting in an accumulation of AKH-containing secretory granules. Additionally, secretory material is stored in large intracisternal granules. Although only a limited part of these large reserves appears to be readily releasable, this strategy allows the adipokinetic cells to comply with large variations in secretory demands; changes in secretory activity do not affect the rate of hormone biosynthesis. AKH-induced lipid release from fat body target cells has revealed a novel concept for lipid transport during exercise. Similar to sustained locomotion of mammals, insect flight activity is powered by oxidation of free fatty acids derived from endogenous reserves of triacylglycerol. However, the transport form of the lipid in the circulatory system is diacylglycerol (DAG) that is delivered to the flight muscles associated with lipoproteins. While DAG is loaded onto the multifunctional insect lipoprotein, high-density lipophorin (HDLp) and multiple copies of the exchangeable apolipoprotein III (apoLp-III) associate reversibly with the expanding particle. The resulting low-density lipophorin (LDLp) specifically shuttles DAG to the working muscles. Following DAG hydrolysis by a lipophorin lipase, apoLp-III dissociates from the particle, regenerating HDLp that is re utilized for lipid uptake at the fat body cells, thus functioning as an efficient lipid shuttle mechanism. Many structural elements of the lipoprotein system of insects appear to be similar to their counterparts in mammals; however, the functioning of the insect lipoprotein in energy transport during flight activity is intriguingly different. PMID- 14529749 TI - Biochemical and immunological adaptation in schistosome parasitism. AB - The objective of this review is to clarify aspects of immunological and biochemical adaptations of schistosomes to their intermediate and final mammalian hosts. Adaptations to the mammalian hosts are traced in relation to cercarial penetration of mammalian skin, glucose transport and metabolism. The unusual ability of schistosome surface membrane to escape immune recognition and damage are reviewed. Moreover, the behavioural changes induced in the intermediate hosts by schistosomes are considered. The evolutionary adaptation to molluscan hosts aims to increase the probability of transmission of the parasite into its mammalian host. This review inspires more hope for further design of anti schistosome drugs through disturbing aspects of biochemical and immunological adaptations in schistosome parasitism. PMID- 14529750 TI - Hormonal regulation of postnatal chicken preadipocyte differentiation in vitro. AB - This study was designed to develop a culture system from the stromal-vascular fraction of chicken adipose tissue that can be used to characterize hormones that promote preadipocyte differentiation. Abdominal adipose tissue was excised from 2 to 4-week-old male broilers (Gallus domesticus) by sterile dissection. The stromal-vascular cell fraction from the adipose tissue was isolated by collagenase digestion, filtration, and subsequent centrifugation. These preadipocytes were seeded in six well culture plates and proliferated to confluency in 10% fetal bovine serum in DMEM/F12 (50:50) medium. At confluency, experiments were initiated to determine hormonal requirements for differentiation. Insulin (100 nM) stimulated expression of citrate lyase and sn glycerol-3-phosphate dehydrogenase relative to lactate dehydrogenase in the presence of 2.5% chicken serum (P<0.05), but not with 10% chicken serum (P>0.05). Triiodothyronine (T(3), 1 nM) and insulin-like growth factor 1 (100 ng/ml) had no effect on differentiation. Dexamethasone (Dex, 1 microM) stimulated differentiation in 2.5 or 10% chicken serum (P<0.05). Insulin, Dex and 2.5% chicken serum stimulated enzymatic differentiation to the extent of 10% chicken serum, but heparin (10 U/ml) addition, in combination with insulin and Dex was necessary to stimulate lipid filling of adipocytes. PMID- 14529751 TI - Coagulant thrombin-like enzymes from the venoms of Brazilian and Peruvian bushmaster (Lachesis muta muta) snakes. AB - Two isoforms of a thrombin-like enzyme designated TLE-B and TLE-P were purified from the venoms of Lachesis muta muta (bushmaster) snakes captured in two different geographical localities, Manaus (Brazil) and Pucallpa (Peru). TLE-B and TLE-P showed Mr values of 44000 and 43000 under reducing conditions on SDS-PAGE, which decreased to 27000 after deglycosylation with N-glycosidase F (PNGase F). The purified proteinases split off fibrinopeptide A rapidly from human fibrinogen and fibrinopeptide B more slowly. In addition, both enzymes released the N terminal peptide (Mr=4572) containing the first 42 residues from the Bbeta-chain. Their specific clotting activities were equivalent to 1000 and 900 NIH thrombin units/mg on human fibrinogen and 526 and 606 NIH thrombin units/mg on bovine fibrinogen for TLE-B and TLE-P, respectively. Kinetic properties of these enzymes were determined using representative chromogenic substrates. Tryptic peptide mapping of the two native enzymes suggested a large degree of structural similarity. Purified rabbit IgG against TLE-B reacted with both enzymes forming a continuous precipitin line on immunodiffusion. Furthermore, Western blot and indirect ELISA were used to compare the antigenic cross-reactivity for both enzymes as well as the venoms of L. muta muta and Bothrops snakes. Incubation of human alpha2-macroglobulin (alpha2-M) with each enzyme at molar ratios of 1:1, 1:2 and 1:4 enzyme:inhibitor resulted in retarding their clotting activities by approximately 12 times, whereas their amidolytic activities were not affected. However, the Mr 180000 subunits of alpha2-M were not cleaved by these enzymes, suggesting that alpha2-M inhibits TLEs by steric hindrance. Similarly, inhibitions of their clotting activities were obtained using high concentrations of rabbit IgG (40 microg, corresponding to molar ratio enzyme:inhibitor of 1:2) against TLE-B. PMID- 14529752 TI - Trafficking of L-triiodothyronine between ovarian fluid and oocytes of rainbow trout (Oncorhynchus mykiss). AB - The study examines the dynamics of thyroid hormone (TH) trafficking between rainbow trout (Oncorhynchus mykiss) oocytes and ovarian fluid (OF) to explore the processes involved in the transfer of hormone to the oocytes. We also examined the effects of enhancing oocyte T(3) content and subsequent embryo survival. Oocytes incubated in OF alone had significant losses of THs within 12 h, whereas the T(3) content of oocytes retained in T(3)-enriched OF (10 and 100 microg ml( 1)) was significantly elevated in a dose-dependant manner within 3 h. When transferred to non-supplemented OF, the T(3) content of the 10 micro ml(-1) treatment group decreased significantly within 24 h with a concomitant significant increase in OF T(3) concentration. Although there was no significant change in the 100 microg ml(-1) treatment group the significant increase in the OF T(3) concentration was evidence for marked T(3) efflux during this period. These findings provide evidence for the independent trafficking of T(3) based on concentration gradients across the oocyte cell membrane, and suggest that it is not vitellogenin-dependent. Fertilization of in ovo T(3)-supplemented oocytes resulted in a small, albeit significant, increase in mortality rate, but there was no significant effect of treatment on embryo growth rates up to the hatching stage of development. PMID- 14529753 TI - Heat- and cold-inducible regulation of HSP70 expression in zebrafish ZF4 cells. AB - Elevated temperature induces a rapid heat shock transcription factor (HSFs) mediated expression of heat shock (hsp) genes. The effect of cold exposure on hsp gene expression has hardly been investigated, although ectothermic animals experience both cold and heat stress. We have previously shown in zebrafish that the expression of hsf1a and a unique isoform hsf1b vary in a tissue-specific manner upon heat stress. In the current study, using a zebrafish (Danio rerio) embryonic cell line (ZF4), we have compared the effects of heat shock (28-->37 degrees C) vs. cold shock (28-->20 degrees C) on the expression of ahsf1a, zhsf1b and hsp70. Concomitantly, the suitability of the ZF4 cells as a model system was verified. The expression pattern of HSP70 proteins following heat or cold exposure is distinct, and the total HSP70 level is upregulated or stable, respectively. Moreover, heat exposure specifically increases the ratio of zhsf1a/b expression (10-fold), whereas cold exposure decreases it to one half. These data suggest that the zhsf1a/zhsf1b ratio is regulated in a temperature dependent manner, and the ratio may be indicative of the stressor-specific HSP70 expression. Furthermore, the response in ZF4 cells upon heat shock resembles the response observed in zebrafish liver and thus, supports the use of this cell line in stress response studies. PMID- 14529754 TI - Cloning of European eel (Anguilla anguilla) FSH-beta subunit, and expression of FSH-beta and LH-beta in males and females after sex determination. AB - The European eel (Anguilla anguilla) is a catadromic teleost species with a complex life cycle, both in sea and freshwater environments. The sex determination phase of gonadal development occurs in a freshwater environment. Polymorphism occurs in increasing rates with respect to gender. While males stop growing at approximately 150 g, females continue to grow to being much larger. In this study, we cloned the cDNA FSH-beta subunit of the European eel (A. anguilla), and measured the mRNA levels of FSH-beta and LH-beta in males and females after sex determination. The FSH-beta subunit cDNA consisted of 1068 bp, encoding a 127 amino acid peptide. A comparison between European and Japanese eels of the FSH-beta amino acid sequence showed 98% similarity. PMID- 14529755 TI - Composition of the surface hydrocarbons from the vitelline membranes of dipteran embryos. AB - Hydrocarbons were the major lipid class extracted by hexane from the vitelline membrane surface of dechorionated eggs of the house fly, Musca domestica, the New World screwworm, Cochliomyia hominivorax, the secondary screwworm, Cochliomyia macellaria, the green bottle fly, Phaenicia sericata, the sheep blow fly, Lucilia cuprina and the Mexican fruit fly, Anastrepha ludens. The length of time the embryos must be exposed to hexane with or without a small amount of alcohol in order to attain permeability was species-dependant. Long-chain n-alkanes comprised the major lipid class removed from vitelline membranes of all species except A. ludens where 2-methylalkanes were the major class. The range in size by the total number of carbon atoms in the hydrocarbons was: C23-C49 in C. hominivorax, C27-C33 in C. macellaria, C24-C35 in L. cuprina, C25-C36 in M. domestica, C25-C33 in P. sericata and C21-C51 in A. ludens. The major hydrocarbon component, expressed as percent of the total hydrocarbons, was n-nonacosane (C29) in C. hominivorax (40%), C. macellaria (43%), L. cuprina (38%), M. domestica (39%) and P. sericata (60%). However, in A. ludens, 2-methyloctacosane (32%) was the major hydrocarbon. Unsaturated hydrocarbons, monoenes (16%) and dienes (11%), were abundant only in A. ludens. Since prior studies indicated that the length of time the embryos must be exposed to hexane with or without a small amount of alcohol in order to attain permeability is species dependant, we suggest that the differences in hydrocarbon composition may contribute to this variation in lipid extractability. PMID- 14529756 TI - Purification and properties of cysteine proteinase inhibitors from rabbit skeletal muscle. AB - Two cysteine proteinase inhibitors, CPI-L and CPI-H, were purified from rabbit skeletal muscle by means of successive extraction with a neutral buffer solution, precipitation at pH 3.7, acetone fractionation and gel permeation on Sephadex G 75 and affinity chromatography on carboxymethyl-papain-Sepharose. The molecular mass of CPI-L was 13 kDa on gel permeation chromatography and SDS-PAGE under reducing conditions and was 15 kDa on SDS-PAGE under non-reducing conditions. The molecular mass of CPI-H was 23 kDa on gel permeation chromatography and it was converted to 13 kDa by SH-reducing agent. Although CPI-H showed single protein band with 13 kDa on SDS-PAGE under reducing conditions, it showed four protein bands with 21, 20, 15 and 13 kDa on SDS-PAGE under non-reducing conditions. Therefore, CPI-H was suggested to have a complicated subunit structure for which S-S bonds and some non-covalent bonds would be responsible. CPI-L and CPI-H were stable in the range of pH 3.0-9.5 and up to 80 degrees C. CPI-L and CPI-H were suggested to inhibit cathepsins B, H and L by a non-competitive mechanism. The inhibition constants (Ki) of CPI-L and CPI-H showed that both CPIs have much higher affinity against cathepsins H and L than against cathepsin B. PMID- 14529757 TI - Fatty acids of astaxanthin esters in krill determined by mild mass spectrometry. AB - Krill is a major source of astaxanthin, which has strong antioxidant activity. Fractions with astaxanthin monoesters and diesters of Antarctic krill Euphausia superba were isolated. Astaxanthin esters were separated by C18-HPLC depending on the number of carbons and double bonds of esterified fatty acid(s). Small amounts of other lipids remained in the samples, but relative molecular masses of carotenoid esters could be measured by field desorption mass spectrometry without fragmentation and interference from contaminant lipids. The fatty acids were determined by calculation of difference between astaxanthin and astaxanthin esters. Only five kinds of fatty acids, dodecanoate, tetradecanoate, hexadecanoate, hexadecenoate and octadecenoate, were detected. Fast atom bombardment mass spectrometry and secondary ion mass spectrometry showed similar spectra. The fatty acid composition in astaxanthin esters was different from those in krill lipids. Therefore, determination of fatty acids in carotenoid esters by a combination of HPLC elution profile and mild mass spectrometry is found to be a useful tool. PMID- 14529758 TI - Temperature dependent characteristics of intestinal glycyl-L-leucine dipeptidase in boreal zone fish. AB - Three kinds of boreal zone fish were investigated for gastrointestinal glycyl-L leucine (GL) dipeptide cleaving activity as a function of feeding stage and seasonal changes. The enzyme activity tested in the perch (Perca fluviatilis L.) intestine increased steadily during digestion and rapidly disappeared after completion. The temperature characteristics and the seasonal changes in dipeptide cleaving activity in pike perch (Stizostedion lucioperca L.) and bream (Abramis brama L.) were studied. In summer, the maximal activities in the pike perch and the bream were found at temperatures of 40 and 30 degrees C, respectively. In winter, the temperature of maximal activity in pike perch fell to only 30 degrees C, whereas no changes were observed in bream. The activation energies in bream and pike perch were several times lower in winter than in summer. Seasonal changes in the dipeptide cleaving activity at low temperature relative to that at the temperature of maximal activity were found. At high temperatures, the stability of the enzyme decreases in winter and increases in summer, but in the presence of a substrate the thermal stability of the enzyme increases both in winter and in summer. In our experiments, we found that in these fish, GL dipeptidase was unstable at 0 and -10 degrees C. PMID- 14529759 TI - Regulation of the rainbow trout (Oncorhynchus mykiss) hepatic beta2-adrenoceptor by adrenergic agonists. AB - The characteristics of hepatic beta(2)-adrenoceptors (AR) were examined in rainbow trout (Oncorhynchus mykiss) chased once per day to exhaustion for up to 7 days or fed the repartitioning agents clenbuterol (CLEN) or ractopamine (RACT) that function in mammals as beta-agonists. A one-day chase and feeding the CLEN for 37 days resulted in a significant 27% and 33% decrease, respectively, in the number of CGP-binding sites (B(max)) with no significant change in affinity (Kd) of hepatic beta(2)-ARs. Despite the significant decrease in beta(2)-AR numbers with CLEN feeding, no significant differences were found for either beta(2)-AR mRNA levels or adenylyl cyclase (ACase) activities. In addition, CLEN displayed only partial agonist activities as it was found to be more effective at blocking isoproterenol-stimulated cAMP production in isolated hepatocytes than stimulating cAMP production. The small affects of RACT may be related to its low active stereoisomer content and low affinity for the trout beta(2)-AR. Agonist regulation of the trout hepatic beta(2)-ARs may involve down-regulation of the receptors without affecting responsiveness. PMID- 14529760 TI - Characterization of the cuticular surface wax pores and the waxy particles of the dustywing, Semidalis flinti (Neuroptera: Coniopterygidae). AB - The adult dustywing, Semidalis flinti Meinander (Neuroptera: Coniopterygidae), begins producing circular-shaped waxy particles after eclosion. The waxy material, which forms the particles, is extruded from individual pores found in clusters on the abdomen. Pores also are present in two rows of three pores on the frontalis and two pores on the first segment of each antennae. The pores have a rosette-like appearance and each pore extrudes dual waxy ribbons. As each ribbon extends a short distance out of the pore, it begins to curl back on itself until the end makes contact with the ribbon. The curled end then breaks free from the extruding ribbon to form the circular waxy particles with fluted edges approximately 2.75-microm diameter. The adults use the particles to cover all parts of their body except for their eyes and appear to lightly coat their antennae. The lipid portion of the particles consists largely of free fatty acids, almost exclusively the 24-carbon fatty acid, tetracosanoic acid. Minor lipid classes are hydrocarbons, fatty alcohols and unidentified material. PMID- 14529761 TI - Differential incorporation of docosahexaenoic and arachidonic acids by the yolk sac membrane of the avian embryo. AB - During avian development, lipoproteins derived from yolk lipid are assembled in the yolk sac membrane (YSM) for secretion into the embryonic circulation. To investigate how yolk polyunsaturated fatty acids, essential for the development of certain tissues, are distributed among the lipid classes of the lipoproteins, pieces of YSM were incubated in vitro with [14C]arachidonic and [14C]docosahexaenoic acids (DHA). There was a marked difference in the partitioning of these two precursors among the lipid classes of the tissue. Of the radioactivity incorporated into total lipid from [14C]-arachidonic acid during 1 h of incubation, 67.3% was esterified as phospholipid and 29.5% as triacylglycerol. In contrast, only 14.6% of the label incorporated from [14C]-DHA was esterified as phospholipid, whereas 73.2% was recovered in triacylglycerol. This pattern of differential partitioning was observed at all time points and across a 20-fold range of fatty acid concentrations. There was no evidence for conversion of the radioactive arachidonic and DHAs to other fatty acids prior to incorporation into tissue lipids. It is suggested that the selective incorporation of yolk-derived DHA into the triacylglycerol of secreted lipoproteins represents part of a mechanism for directing this polyunsaturate to particular embryonic tissues. PMID- 14529762 TI - Captive roe deer (Capreolus capreolus) select for low amounts of tannic acid but not quebracho: fluctuation of preferences and potential benefits. AB - Browsing ruminants have been shown to tolerate a certain amount of tannins in their natural diet, and preference trials with captive roe deer (Capreolus capreolus) have suggested an active selection for a low dose of hydrolysable tannins. In this study, we investigated the preference patterns for tannic acid, a source of hydrolysable tannins, and quebracho, a source of condensed tannins, in a series of preference trials with captive roe deer over time, using a pelleted feed that differed only in the respective tannin content. Additionally, two groups of four hand-raised roe deer fawns were fed either a control or a 3% tannic-acid containing diet and physiological parameters were compared after 7.5 months. There were large differences in preference patterns between the individual roe deer groups; quebracho was mostly avoided, whereas tannic acid was actively included in the diet in differing, low proportions. However, one group consistently preferred the quebracho diet over both the control or the tannic acid diet. For the tannic acid, the preference pattern often revealed an initial period of high preference, followed by a stable period of a moderate preference. The fawns on the tannic acid diet had a lower pellet intake and a higher relative mass gain than the fawns on the control diet; differences in salivary tannin binding capacity and in blood antioxidant status were below significance. These results are the first indications of potential benefits of a low-dose tannin diet, which need further confirmation. The results of the preference trials demonstrate that the time pattern of tannin intake is not constant, and pose the question about the validity of short-term preference trials in general. PMID- 14529763 TI - Near simultaneous computed tomography image-guided stereotactic spinal radiotherapy: an emerging paradigm for achieving true stereotaxy. AB - PURPOSE: To report treatment setup data from an emerging technique using near simultaneous computed tomography (CT) image-guided stereotactic radiotherapy for the treatment of spinal and paraspinal tumors. METHODS AND MATERIALS: A targeting system that integrates a CT-on-rails scanner with a linear accelerator (LINAC) was evaluated in the lead-in portion of a Phase I/II protocol for treating patients with paraspinal metastases. Patients were immobilized in supine position by a moldable body cushion vacuum wrapped with a plastic fixation sheet. Planning CT and immediately repeated CT were performed on the LINAC/CT-on-rails unit to assess respiratory-related vertebral body motion. Coplanar intensity-modulated radiotherapy (IMRT) using 7-9 beams was used to deliver 30 Gy in five fractions to the target volume, while limiting the spinal cord dose to <10 Gy. Pretreatment CT scans were fused with the planning CT scans to determine the correct target isocenter by accounting for any translational and roll (axial) rotational discrepancies from the planning CT. (Corrections caused by yaw and pitch rotations have not yet been implemented.) The reproducibility of the treatment isocenter as compared with the planned isocenter was measured with digitally reconstructed radiographs (DRRs), portal film imaging, and immediate post treatment verification CT scans. Phantom measurements were taken for dose verification for each IMRT plan. RESULTS: Based on a total of 36 CT scans (3 for planning, 3 for respiration study, 15 pretreatment, and 15 post-treatment) from 3 patients, no respiration-associated vertebral body motion was seen. A comparison of the corrected daily anterior-posterior (AP) and lateral (LAT) digital portal images with the planning AP and LAT DRRs confirmed that the isocenter setup accuracy for the 15 treatments was within 1 mm of the planning isocenter. The results from the immediate post-treatment CT scans reconfirmed the findings from the portal images and verified the absence of spinal movement during the treatment. The ion-chamber measurement for the high-dose region was within 2% of the planning dose for three patient treatment plans. Film dose measurement in an IMRT quality assurance phantom demonstrated good agreement from 90% to 30% isodose lines between the planned and measured results. CONCLUSION: Preliminary experience suggests that the near-simultaneous CT image-guided verification technique can be used as a new platform technology for extracranial applications of stereotactic radiotherapy and radiosurgery to spinal and paraspinal tumors. PMID- 14529764 TI - RTOG 94-06: is the addition of neoadjuvant hormonal therapy to dose-escalated 3D conformal radiation therapy for prostate cancer associated with treatment toxicity? AB - PURPOSE: This study determines the effect on toxicity of adding neoadjuvant hormonal therapy (NHT) to three-dimensional conformal radiation therapy (3D-CRT) in RTOG 94-06. METHODS AND MATERIALS: Between August 1994 and February 2000, 583 eligible prostate cancer patients enrolled on the first 3 dose levels of RTOG 94 06, a Phase I/II dose escalation 3D-CRT trial. Two hundred and seven men initiated hormonal therapy (HT) between 2 to 3 months before 3D-CRT, and completed all HT no longer than 3 months after radiotherapy. Thirty-three patients receiving longer-duration HT were excluded. The 547 patients were treated at dose level I (68.4 Gy), level II (73.8 Gy), or level III (79.2 Gy). All dose prescriptions were to the minimum isodose surface encompassing the planning target volume (dose levels I and II) or the clinical target volume (dose level III). Men were stratified into three risk groups according to their relative risk of seminal vesicle invasion: <15% (Group 1) vs. >15% (Group 2), or to T stage (T1, 2 vs. T3 tumors [Group 3]). In Group 2 patients, there was a clinical target volume reduction to treat only the prostate after delivery of 55.8 Gy to a planning target volume including the seminal vesicles. All HT consisted of a luteinizing hormone-releasing hormone agonist with or without a nonsteroidal anti-androgen. RESULTS: On univariate analysis, NHT significantly increased the likelihood of Grade 2 acute genitourinary (GU) complications (22% to 32%, p = 0.009). Hormonal therapy did not have a significant univariate effect on any other acute or late toxicity. On multivariate analysis, the percent of the bladder (< or =30% vs. >30%) receiving > or = the reference dose (68.4 Gy, 73.8 Gy, or 79.2 Gy) (p = 0.0009, relative risk = 2.07, confidence interval: 1.88 2.28) was a significant predictor of acute GU effects. Although NHT was not significant in itself, in the multivariate analysis its interaction with baseline urinary status was an important factor (p = 0.011, relative risk = 4.31, confidence interval: 1.68-5.29). CONCLUSION: Neoadjuvant HT did not show an independent effect on the risk of side effects after 3D-CRT in patients treated on RTOG 94-06. However, this combined modality therapy significantly increased the risk of acute GU effects compared to 3D-CRT alone in men with poor baseline urinary function. PMID- 14529765 TI - Influence of follow-up bias on PSA failure after external beam radiotherapy for localized prostate cancer: results from a 10-year cohort analysis. AB - PURPOSE: To estimate the biases inherent in prostate cancer outcome that arise from different failure end points and variations in follow-up time and intensity using a cohort of men with long follow-up. METHODS AND MATERIALS: The study cohort consisted of 205 men with T1-T2N0-Nx prostate cancer treated with conventional radiotherapy between 1991 and 1993. The median follow-up was 103 months. Outcome was assessed using different definitions of biochemical failure, including the American Society for Therapeutic Radiology and Oncology (ASTRO) consensus criteria and the "nadir plus two" criteria (any rise of 2 ng/mL greater than the current nadir). Patient subgroups were created according to where patients had received their last 2 years of follow-up. Patients were also stratified according to whether they were initially present in the departmental database (under regular surveillance) or were uncovered after more vigorous investigation (previously "lost to follow-up"). RESULTS: In this series with maximized follow-up, the 10-year biochemical disease-free survival rate did not change significantly with varying definitions of failure, 49% and 45% for ASTRO and "nadir plus two" criteria, respectively. Patients followed by outside physicians (n = 99) were faring better at 10 years than those followed at the treating institution by either their radiation oncologist (n = 50) or their medical oncologist or urologist (n = 52). This was by all measures of outcome, including overall survival, and metastasis-free survival. Patients previously lost to follow-up (n = 43) who were tracked down also appeared to be doing better than those on our database for whom information had been readily available (n = 161). This, however, may have been an artifact of the ASTRO criteria, which underestimates failure when insufficient prostate-specific antigen values are available. CONCLUSION: The ASTRO definition of failure underestimates late failure. This bias may be compensated for by the use of cohorts with long follow up or the use of the "nadir plus two" definition of failure. The use of institutional prostate cancer databases may overestimate failure rates because patients followed outside of the treating institution fared better with respect to both survival and biochemical recurrence. Vigorous attempts to obtain follow up beyond the hospital walls may correct this bias. PMID- 14529766 TI - Improvement in relapse-free survival throughout the PSA era in patients with localized prostate cancer treated with definitive radiotherapy: year of treatment an independent predictor of outcome. AB - PURPOSE: In patients treated with radical prostatectomy in the prostate-specific antigen (PSA) era, it has been demonstrated that the year of treatment in the PSA era is associated with better pathologic parameters and outcomes, independently of other well-recognized parameters such as clinical stage, pretreatment PSA level, or Gleason score. The purpose of the present study was to study a similar phenomenon with definitive radiotherapy (RT). METHODS AND MATERIALS: The inclusion criteria were as follows: clinical Stage T1-T2, available pretreatment PSA level and biopsy Gleason score, treatment delivered before January 2000 with standard fractionation external beam radiotherapy to at least 70 Gy, no adjuvant androgen deprivation (AD), all neoadjuvant AD limited to < or =6 months, and a minimum of 3 years of PSA follow-up. A total of 467 cases treated between January 1986 and December 1999 were included. Short-course AD in the adjuvant or neoadjuvant setting for < or =6 months was given in 124 cases (27%). The median radiation dose was 74 Gy (range 70.0-78.0). A conformal technique was used in 293 cases (63%). The median follow-up was 62 months (range 37-189). A total of 4931 follow-up PSA levels were available for analysis (average 11 per patient). A multivariate analysis for factors affecting biochemical relapse-free survival rates using the proportional hazards model was performed for all cases using the following variables: age (continuous variable), race (black vs. white), clinical T stage (T1-T2a vs. T2b-T2c), pretreatment PSA (continuous variable), biopsy Gleason score (continuous variable), use of AD (yes vs. no), radiation dose (continuous variable), and year of treatment (continuous variable: 1986-1999). RESULTS: The projected 8-year biochemical relapse-free survival rate was 74%. The projected 5-year biochemical relapse-free survival rate for the 143 patients treated in the 1986-1995 period was 58% vs. 82% for the 324 patients treated in the 1996-1999 period (p <0.001). The difference was attributable to a multitude of factors (earlier stage cancer, higher radiation doses, shorter follow-up). To study the confounding effects of these factors on the year of therapy, a multivariate analysis was performed. The multivariate analysis revealed the initial PSA level (p <0.001), Gleason score (p <0.001), RT dose (p = 0.045), and year of treatment (p <0.001) to be independent predictors of outcome. Age (p = 0.41), race (p = 0.14), T stage (p = 0.10), and use of AD (p = 0.58) were not. CONCLUSION: When controlling for tumor, treatment, and follow-up parameters, the year in which RT was performed was still an independent predictor of outcome, consistent with observations made for radical prostatectomy patients. This indicates a more favorable presentation of localized prostate in current years probably related to a combination of factors such as screening and increased patient awareness leading to earlier diagnosis. Outcome predictions should be based on contemporaneous series. PMID- 14529767 TI - Evaluation of ultrasound-based prostate localization for image-guided radiotherapy. AB - To evaluate the use of the ultrasound-based BAT system for daily prostate alignment. Prostate alignments using the BAT system were compared with alignments using radiographic images of implanted radiopaque markers. The latter alignments were used as a reference. The difference between the BAT and marker alignments represents the displacements that would remain if the alignments were done using ultrasonography. The inter-user variability of the contour alignment process was assessed. On the basis of the marker alignments, the initial displacement of the prostate in the AP, superoinferior, and lateral direction was -0.9 +/- 3.9, 0.1 +/- 3.9, and 0.2 +/- 3.4 mm respectively. The directed differences between the BAT and marker alignments in the respective directions were 0.2 +/- 3.7, 2.7 +/- 3.9, and 1.6 +/- 3.1 mm. The occurrence of displacements >/=5 mm was reduced by a factor of two in the AP direction after the BAT system was used. Among eight users, the average range of couch shifts due to contour alignment variability was 7, 7, and 5 mm in the antero-posterior (AP), superoinferior, and lateral direction, respectively. In our study, the BAT alignments were systematically different from the marker alignments in the superoinferior, and lateral directions. The remaining random variability of the prostate position after the ultrasound-based alignment was similar to the initial variability. However, the occurrence of displacements >/=5 mm was reduced in the AP direction. The inter user variation of the contour alignment process was significant. PMID- 14529768 TI - Biochemical outcomes after prostate brachytherapy with 5-year minimal follow-up: importance of patient selection and implant quality. AB - PURPOSE: A prostate brachytherapy program was initiated in 1990, when comparatively little was known of the relative importance of disease- and treatment-related factors on outcome. Patients treated during the first 6 years of the program were analyzed to determine the value of patient selection and implant quality on biochemical control. METHODS AND MATERIALS: We treated 243 patients with clinically localized prostate cancer with radioactive seed implantation and underwent 1-month CT-based dosimetric analysis. Follow-up ranged from 61 to 135 months (median 75). The Gleason score was < or =6 in 78% (n = 189), 7 in 14% (n = 35), and 8-10 in 8% (n = 19). The initial prostate-specific antigen (PSA) level was < or =10 ng/mL in 61% (n = 149), 10.1-20 ng/mL in 26% (n = 63), and >20 ng/mL in 13% (n = 31). The disease stage was T2a or less in 49% (n = 120), and Stage T2b-T2c in 51% (n = 123). A real-time ultrasound-guided technique was used with (125)I (n = 138) and (103)Pd (n = 105) isotopes. No patient underwent external beam radiotherapy as part of their primary treatment. Of the 243 patients, 60% also received hormonal ablation for at least 3 months before and 2-3 months after seed implantation. All patients included underwent a 1-month CT-based dosimetric analysis. The implant dose was defined as the dose delivered to 90% of the prostate volume on postimplant dosimetry (D(90)). On the basis of prior dose-response analyses, patients were retrospectively grouped into optimal D(90) ((125)I > or =140 Gy Task Group 43 or (103)Pd >/=100 Gy) and suboptimal D(90) ( (125)I <140 Gy or (103)Pd <100 Gy) dose groups. Biochemical failure was defined using the American Society for Therapeutic Radiology Oncology definition. RESULTS: Disease-related factors, including initial PSA level, Gleason score, and stage, were significant predictors of biochemical failure. The actuarial 8-year freedom from biochemical failure (bFFF) rate was 80% for those with a PSA level < or =10 ng/mL, 86% for PSA 10.1-20 ng/mL, and 45% for PSA >20 ng/mL (p = 0.0019). Patients with a Gleason score of < or =6 had an 8-year bFFF rate of 81% vs. 67% for those with Gleason score 7 and 53% for those with Gleason score 8-10 (p = 0.0003). Patients with Stage T2a or less had an 8-year bFFF rate of 85% compared with 69% for those with Stage T2b-T2c (p = 0.013). The 8-year bFFF rate was 88% for low-risk patients (Stage T2a or less, Gleason score < or =6, and initial PSA level < or =10 ng/mL; n = 75), 81% for moderate-risk patients (Stage T2b or Gleason score 7 or initial PSA level >10.1-20 ng/mL; n = 70), and 65% for high-risk patients (two or more moderate-risk features or Gleason score > or =8 or initial PSA level >20 ng/mL; n = 98; p = 0.0009). Patients with optimal dose implants (n = 145) had an 8-year bFFF rate of 82% compared with 68% for those with suboptimal dose implants (n = 98; p = 0.007). Hormonal therapy did not significantly affect biochemical failure (p = 0.27). In multivariate analysis, the statistically significant variables included initial PSA level (p <0.0001), Gleason score (p = 0.024), and dose group (p = 0.046). Because our current practice limits implantation alone to low-risk patients, an analysis of this subgroup was undertaken to validate the importance of dose. In the optimal dose group, low-risk patients had an 8-year bFFF rate of 94% vs. 75% for the low-risk patients in the suboptimal dose group (p = 0.02). CONCLUSION: With minimal follow up of 5 years, these data continue to support the use of implantation alone in low-risk prostate cancer patients and demonstrate the importance of implant quality (dose) in achieving optimal outcomes. Low-risk patients who receive an optimal dose implant have a 94% bFFF rate at 8 years. PMID- 14529769 TI - Efficacy of irradiation and external hyperthermia in locally advanced, hormone refractory or radiation recurrent prostate cancer: a preliminary report. AB - PURPOSE: To present a preliminary report on the feasibility, efficacy, and toxicity of irradiation (RT) and hyperthermia (HT) in patients with locally advanced, hormone-refractory prostate cancer (LAHRPC) who may or may not have received prior RT. METHODS AND MATERIALS: Between 1997 and 2002, 13 consecutive patients with LAHRPC or RT-recurrent prostate cancer were treated with RT and HT on a Phase I-II protocol. Eight patients had RT-recurrent LAHRPC (Group A) and 5 had LAHRPC without prior RT (Group B). All patients had large and clinically symptomatic tumors. The median RT dose was 39.6 Gy and 66.6 Gy in Groups A and B, respectively. External deep HT was delivered using a BSD-2000 Sigma-60 applicator. The median number of HT treatments was 8 in group A and 10 in group B. RESULTS: The median follow-up was 14 and 13 months for Groups A and B, respectively. All patients achieved a complete or partial response (CR/PR) and complete palliation of symptoms. Eleven patients had follow-up CT scans that demonstrated a CR in six and a PR in five. Two patients, who died of metastasis, did not have CT scans and had a PR on digital rectal examination. Two patients demonstrated a biochemical CR. The median duration of the CR/PR among Group A patients was 12 months after therapy. Three patients in Group A developed tumor recurrence at 9, 17, and 27 months after repeat RT to doses of 39.6, 36, and 50 Gy, respectively. At last follow-up, no Group B patient developed local recurrence. Grade 1-2 rectal bleeding was noted in 3 patients. RT and HT were generally well tolerated by all patients who had not previously undergone RT. Of the 8 patients who had, 6 (75%) tolerated retreatment well with minimal or no complications. Two patients in the repeat RT group had severe complications. One patient with lymphoma and factor XI deficiency developed Grade 4 hemorrhagic cystitis. Another previously irradiated patient developed a rectovesical fistula 4 months after retreatment, after disappearance of a large, invasive, and necrotic tumor. CONCLUSION: This preliminary report demonstrates the feasibility and efficacy of RT and HT in patients with LAHRPC, who may or may not have received prior RT. Presently, such patients who have undergone previous RT have no effective treatment options. RT and HT were generally well tolerated by patients who were not previously undergone RT. Of those who had been, most (6 of 8) tolerated retreatment well with minimal or no complications. The high-risk factors for treatment- and tumor regression-related side effects include the presence of large necrotic tumors, previous RT with a large dose/fraction, and the presence of bleeding disorders. Despite the size of these large tumors, RT and HT resulted in significant tumor shrinkage, rapid serum prostate-specific antigen decline, durable treatment responses, and durable palliation of symptoms. Additional clinical studies are warranted. PMID- 14529770 TI - RTOG 97-06: initial report of a phase I-II trial of selective bladder conservation using TURBT, twice-daily accelerated irradiation sensitized with cisplatin, and adjuvant MCV combination chemotherapy. AB - To examine combination cisplatin and twice-daily accelerated irradiation (RT) after aggressive transurethral resection of bladder tumor (TURBT) in an attempt to preserve the bladder and to determine the likelihood that patients who complete this regimen could then complete three cycles of methotrexate, cisplatin, vinblastine (MCV) chemotherapy. Between 1998 and 2000, 52 patients with Stage T2-T4aN0M0 disease, from 17 institutions, were entered into the trial. Forty-seven patients were deemed eligible; the planned accrual was 40. Of the 46 patients, 68% were >60 years old, 70% were men, and 96% had a Karnofsky score >/=90. The clinical T stage was T2 in 66%, T3a in 25%, and T3b in 9%. The median follow-up at the time of analysis was 26 months. The protocol required TURBT within 6 weeks of the initiation of induction therapy. Induction treatment involved 13 days of concomitant boost RT, 1.8 Gy to the pelvis in the morning followed by 1.6 Gy to the tumor 4-6 h later. For sensitization, cisplatin (20 mg/m(2)) was given on the first 3 days of each treatment week. Three to four weeks after induction, patients were evaluated cystoscopically for residual disease. Patients whose biopsies and cytologic evaluations showed no disease completed consolidation chemoirradiation. Patients with residual tumor went on to cystectomy. After either consolidation or cystectomy, patients were to complete three cycles of MCV chemotherapy. Of the 47 patients, 45% completed all phases of the protocol treatment with minor, or no, deviations. Five patients refused either the postinduction evaluation or cystectomy and 6 refused adjuvant chemotherapy. The CR rate after induction therapy was 74%. For 2 patients, residual disease after induction was limited to positive cytologic findings, and for 8 patients, biopsy of the primary site revealed persistence. Of the 8 cystectomy patients, 2 had no evidence of disease in the bladder at pathologic review of the surgery specimen. Grade 3 toxicity related to chemotherapy was observed in 11% of patients during both induction and consolidation, and in 41% during adjuvant chemotherapy. A total of 8 patients (36% of those receiving adjuvant chemotherapy) went on to develop Grade 4 neutropenia or thrombocytopenia during additional adjuvant chemotherapy. Grade 3 toxicity due to RT was seen in 4% and 0% of patients during induction and consolidation, respectively. One patient developed Grade 4 hydronephrosis during consolidation. The projected 36 month value for locoregional failure, distant metastasis, overall survival, and bladder-intact survival was 27%, 29%, 61%, and 48%, respectively. After aggressive TURBT, twice-daily accelerated RT initiated in concomitant-boost format is well tolerated and results in a rate of complete response (74%) similar to that in previous bladder-sparing trials. The projected 2-year values for locoregional control, bladder-intact survival, and overall survival were also consistent with previously reported trials of bladder-sparing treatment. With only 45% of patients completing three cycles of MCV, this form of adjuvant chemotherapy appears to be poorly tolerated by most patients. PMID- 14529771 TI - Potential of intensity-modulated radiotherapy to escalate doses to head-and-neck cancers: what is the maximal dose? AB - PURPOSE: To investigate the potential of intensity-modulated radiotherapy (IMRT) to escalate doses to head-and-neck cancer and find the maximal dose that could be prescribed to the target volume with IMRT while doses to critical organs were maintained at their currently acceptable levels. The secondary goal was to search for limits in current IMRT technology. METHODS AND MATERIALS: For a group of 12 head-and-neck cancer patients with different tumor locations and shapes, we performed IMRT planning using a simultaneous integrated boost strategy, that is, the gross tumor volume (GTV), clinical target volume (CTV), and electively treated nodes were treated simultaneously at different dose levels. The critical structures involved in the treatment field that needed to be spared included the brainstem, spinal cord, and parotid glands, depending on the disease site. Nine coplanar 6-MV photon beams were used for planning with the IMRT system developed at our institution, and dose-volume criteria were used for optimization. By varying the optimization parameters, we gradually increased the dose to the GTV while keeping the dose to the critical structures at less than the acceptable tolerance level. The criteria for accepting the plan included the following: (1) the prescription dose to the GTV had to cover 99% of the volume, and the dose homogeneity of the GTV needed to be <10%; (2) the prescription to the CTV (which was set either at 60 Gy or 10 Gy less than that of the GTV) had to cover 95% of the volume, and the same amount of normal tissue outside the CTV received the CTV prescription dose as in the current acceptable plan; (3) the prescription to the electively treated lymph nodes needed to cover 90% of the volume; and (4) the maximal dose to the brainstem and spinal cord had to be <55 Gy and 45 Gy, respectively. For parotid glands, the dose needed to be as low as possible without compromising the target doses. The deliverable plans as determined by the actual multileaf collimator leaf sequences were used for the final evaluation. To verify that the acceptable plans were deliverable, the experimental measurements of planar dose distribution were performed in phantom with film. RESULTS: The maximal dose to the GTV varied from 86 to 176 Gy if the CTV dose increased with the GTV dose. It was reduced to 76-82 Gy if the CTV dose was kept at 60 Gy. The competing criteria usually are the requirements of the tolerance doses to the critical organs and target dose homogeneity, not the target prescription dose. Using more beams only increased the dose marginally. The results could change significantly if a different set of criteria for the plan evaluation were used. Dosimetric measurements confirmed that such a high dose and dose gradient could be delivered accurately with dynamic multileaf collimators. Statistical analyses showed no significant correlations between the maximal doses and the number of GTVs and volume of GTVs and CTVs. CONCLUSION: Doses to head-and-neck cancers with simultaneous integrated boost IMRT can be escalated to a greater level than currently prescribed clinically. The limit of IMRT in head-and-neck cancer has not been reached at the current prescription level of 70 Gy. Such high total and fractionated doses should be carefully evaluated before being prescribed clinically. PMID- 14529772 TI - P105 as a prognostic indicator in patients irradiated for locally advanced head and-neck cancer: a clinical/laboratory correlative analysis of RTOG-9003. AB - PURPOSE: In a previous retrospective study, p105 AD, a proliferation-associated nuclear antigen density (AD), was found to be an independent prognostic factor for patients irradiated for locally advanced head-and-neck cancer. We sought to confirm this finding by analyzing patients entered on RTOG 9003, a Phase III randomized trial of altered fractionation radiotherapy. METHODS AND MATERIALS: Paraffin blocks of pretreatment biopsies of the primary tumor of patients with Stage III or IV squamous cell carcinoma of the oral cavity, oropharynx, or supraglottic larynx, or Stage II squamous cell carcinoma of the hypopharynx or base of tongue entered on RTOG 9003 were prospectively collected at patient entry. From these paraffin blocks, areas of tumor were selected based on histologic examinations and sectioned. Nuclear suspensions were then prepared and processed for p105 antibody and DNA staining. Flow cytometric quantification of p105 labeling indices and DNA content were then performed for correlation with local-regional control and survival. RESULTS: Paraffin blocks of tumor biopsies from 457 of 1073 patients entered were available for p105 determination. There was no significant difference in pretreatment characteristics between patients who had paraffin blocks available or not available. The median (range) of p105 labeling index (LI-C), p105 labeling index of cells in S phase (p105 LI-S), and p105 AD were 56 (range: 6-99), 8.255 (range: 0.913-23), and 67 (range: 5-364), respectively. Multivariate analysis of prognostic factors showed that T stage, N stage, Karnofsky performance status, and fractionation schedule were significant for local-regional control (p < 0.0001, 0.0011, <0.0001, and 0.007, respectively) and T stage, N stage, Karnofsky performance status, and tumor grade were significant for survival (p = 0.018, 0.002, <0.0001, and 0.0058, respectively). Neither p105 LI-C nor p105 LI-S nor p105 AD nor DNA ploidy was significant for local-regional control or survival. CONCLUSION: p105 labeling indices, antigen density, and DNA ploidy do not predict the outcome of patients irradiated for advanced squamous cell carcinomas of the head and neck. PMID- 14529773 TI - Postoperative radiotherapy in squamous cell carcinoma of the oral cavity: the importance of the overall treatment time. AB - PURPOSE: To test the hypothesis that (1) the distinction between intermediate- and high-risk patients by clustering different prognostic factors results in a significant difference in treatment outcome and (2) a shorter interval between surgery and radiotherapy and shorter overall treatment times of radiation (OTTRT) result in higher rates of locoregional control (LRC). METHODS AND MATERIALS: Included were patients (n = 217) with previously untreated squamous cell carcinoma of the oral cavity treated with radical surgery and postoperative radiotherapy. Patients with extranodal spread or microscopic residual disease and patients with two or more other risk factors (i.e., N2b-N3, >1 nodal level involved, perineural growth, or stage T3-T4) were classified as high-risk patients. Patients with only one other risk factor were classified as intermediate risk. RESULTS: In the intermediate-risk group, the 3-year LRC was 87% as compared with 66% in the high-risk group (p = 0.0005). No association was found between interval and LRC. However, the OTTRT was significantly associated with LRC. The 3-year LRC was 87%, 75%, 69%, and 51% when the OTT was <6 weeks, 6 7 weeks, 7-8 weeks, and >8 weeks, respectively (p = 0.0004). The 3-year overall survival (OS) in the intermediate risk patients was 74% compared with 50% in the high-risk group (p = 0.0014). A significant association was also found between the OS and OTTRT. The OS increased from 50% when the OTTRT was >8 weeks to 74% when the OTT was <6 weeks (p = 0.006). Similar results were found with regard to the disease-free survival (DFS). In the multivariate analysis, both risk group and OTT were significantly associated with LRC, DFS, and OS. No significant interaction term was present between these two factors, which means that the OTT was of importance both for the high-risk and the intermediate-risk patients. CONCLUSION: In the subset of patients with carcinoma of the oral cavity, the classification of high- and intermediate-risk patients by clustering a number of prognostic factors provides important prognostic information regarding LRC, DFS, and OS. The OTT was the most important prognostic factor both in the high-risk and intermediate-risk patients. Reducing the OTT to 6 weeks or less is a rather simple measure to achieve a considerable improvement of the outcome of treatment in this category of patients. PMID- 14529774 TI - Radiation dose escalation in limited-stage small-cell lung cancer. AB - PURPOSE: To review the treatment outcomes of limited-stage small-cell lung cancer (LS-SCLC) patients treated with > or =50 Gy of radiation at Massachusetts General Hospital (MGH) between 1987 and 2000 and to assess for evidence of a continuation of a radiation dose response. METHODS AND MATERIALS: The MGH cancer registry was searched for SCLC patients treated with radiotherapy between 1987 and 2000. Records of LS-SCLC patients treated with curative intent and radiation doses > or =50 Gy at MGH were reviewed. Surgical patients were excluded. RESULTS: Eighty four LS-SCLC patients were treated with radiotherapy at MGH between 1987 and 2000. Of the 84 patients, 54 (64%) met the inclusion criteria; 30 patients (56%) in this study died, and 4 (7%) were lost to follow-up. The median follow-up of the surviving patients was 42 months. The median overall survival was 29 months. The 2- and 5-year survival rate was 64% and 47%, respectively. The local control rate at 3 years was 78%. CONCLUSION: The overall survival, local control, and disease-free survival rates for LS-SCLC patients treated with > or =50 Gy of radiation compare favorably with historical data. These findings suggest a continuation of the radiation dose-response curve in LS-SCLC. This further supports the need for appropriately powered, Phase III, prospective randomized trials in radiation dose escalation or radiation dose intensification for LS SCLC. PMID- 14529776 TI - Digital fluoroscopy to quantify lung tumor motion: potential for patient-specific planning target volumes. AB - PURPOSE: To apply digital fluoroscopy integrated with CT simulation to measure lung tumor motion and aid in the quantification of individualized planning target volumes. METHODS AND MATERIALS: A flat panel digital fluoroscopy unit was modified and integrated with a CT simulator. The stored fluoroscopy images were overlaid with digitally reconstructed radiographs, allowing measurement of the observed lung tumor motion in relation to the corresponding contours on the static digitally reconstructed radiographs. CT simulation and digital fluoroscopy was performed on 10 patients with non-small-cell lung cancer. Actual tumor motion was measured in three dimensions using the overlaid images. RESULTS: Combining the dynamic data with digitally reconstructed radiographs allowed the tumor shadow from the fluoroscopy to be tracked in relation to the CT lung tumor contour. For all patients, the extent of tumor motion in three dimensions was unique. The motion was greatest in the superoinferior direction and minimal in the AP and lateral directions. CONCLUSION: We have developed a tool that allows CT simulation to be combined with digital fluoroscopy. Quantitative evaluation of the tumor motion in relation to the CT plan allows for customization of the planning target volume. The variability observed clearly demonstrates the need to generate patient-specific internal motion margins. PMID- 14529775 TI - Shifting from hypofractionated to "conventionally" fractionated thoracic radiotherapy: a single institution's 10-year experience in the management of limited-stage small-cell lung cancer using concurrent chemoradiation. AB - PURPOSE: To perform a retrospective review of a single institution's 10-year experience in treating limited-stage small-cell lung cancer (LS-SCLC) with a concurrent chemoradiation regimen modeled after the experimental arm of a randomized National Cancer Institute of Canada trial in which hypofractionated radiotherapy started with cycle 2 of chemotherapy. We then looked at the impact on patient outcomes of changing the RT during the course of the decade to a "conventionally" (2 Gy) fractionated regimen, with a focus on toxicity and survival rates. METHODS AND MATERIALS: Between 1989 and 1999, 215 LS-SCLC patients received six cycles of chemotherapy consisting of cyclophosphamide, doxorubicin, and vincristine alternating with etoposide and cisplatin every 3 weeks. Thoracic RT was administered concurrently with etoposide and cisplatin (at cycle 2 or 3) only and consisted of either 40 Gy in 15 fractions for 3 weeks or 50 Gy in 25 fractions for 5 weeks. RT fields encompassed gross and suspected microscopic disease with 2-cm margins. Prophylactic cranial irradiation (PCI) was offered to complete responders according to clinician preference. RT interruption during concurrent chemoradiation was used as the "marker" for treatment toxicity. The analysis compared the RT schedules for differences in toxicity, survival, and recurrence patterns. RESULTS: The overall survival rate for 215 patients at 2 and 5 years was 22.7% and 7.2%, respectively, with a median survival of 14.7 months. Thoracic RT consisted of 40 Gy in 3 weeks for 122 patients (57%) and 50 Gy in 5 weeks for 92 patients (43%). PCI was administered to 21 (44%) and 47 (56%) patients receiving 40 Gy and 50 Gy, respectively. The patient- and treatment related variables were comparable between the two cohorts treated with the different RT prescriptions. RT interruptions during concurrent chemoradiation were recorded in 56 cases (26%), with a median duration of 5 days (range 1-18). No differences in treatment-related toxicity rates were demonstrated between the two dose cohorts (p = 0.35). The overall and disease-free survival rates (patients stratified by PCI use) at 5 years for the 40- and 50-Gy schedules were 14.3% and 12.0% (p = 0.71) and 20.7% and 22.2% (p = 0.76), respectively. Sites of first failure were recorded in 132 patients (61%). Comparing the 40-Gy and 50-Gy cohorts, the rate of any first relapse was 40% vs. 42% and the chest as the first relapse site was 34% vs. 45% (patients stratified by PCI use), respectively. The brain failure rate reflected PCI use and was not related to the thoracic RT schedule. CONCLUSION: Changing from a hypofractionated to a conventionally fractionated RT thoracic prescription did not alter outcomes because the survival, thoracic control, and toxicity rates were statistically similar. This suggests that the hypofractionated schedule remains practicable and should be considered in the setting of randomized clinical trials. In view of the benefits that accelerated schedules provide for both patients and cost containment, clinicians may opt to use this tolerable regimen in managing LS-SCLC. Regarding the future development of novel chemoradiation programs, the most critical factor in ensuring improved outcomes for LS-SCLC may be limiting the duration of RT and overall treatment time. PMID- 14529777 TI - The value of a positive margin for invasive carcinoma in breast-conservative treatment in relation to local recurrence is limited to young women only. AB - PURPOSE: To identify the importance of positive margins for invasive carcinoma on local control in patients treated with breast-conservative treatment (BCT). METHODS AND MATERIALS: A total of 1752 BCT with known margins were analyzed. Fifty-five patients had a second BCT, leaving 1697 patients for analysis. The margins were positive in 193/1752 BCT (11%). The median follow-up was 78 months. RESULTS: The 5- and 10-year local recurrence rates (LRR) were 3.1% and 6.9%, respectively, for negative margins vs. 5.6% and 12.2% for positive margins. A statistical interaction between age category and margin status was noted in relation to disease-free survival (DFS) and local relapse-free survival. The 5 year LRR for women < or =40 years was 8.4% for negative margins and 36.9% for positive margins (p = 0.005). In a multivariate analysis, a positive margin was significant. The 5-year LRR for women >40 years was 2.6% for negative and 2.2% for positive margins. The 5-year DFS for women 40 years was 84.3% for positive and 87.2% for negative margins. CONCLUSION: Women < or =40 years are a special category of patients in breast cancer. Women < or =40 years must have negative margins for invasive carcinoma when treated with BCT. Minimum surgery for an optimal cosmetic result followed by irradiation, even with microscopic positive margins for invasive carcinoma, yields excellent results with regard to local control in patients older than 40 years. PMID- 14529778 TI - Lymph nodes in the irradiated field influence the yield of radiation-induced chromosomal aberrations in lymphocytes from breast cancer patients. AB - PURPOSE: To measure chromosomal aberrations in blood lymphocytes from breast cancer patients treated with radiotherapy after quadrantectomy or tumorectomy. METHODS AND MATERIALS: Twenty-two breast cancer patients treated with breast conserving surgery and radiation were evaluated. Adjuvant chemotherapy was also given to 9 patients. Blood samples were obtained before radiotherapy, after about one-half of the fractions, and at the end of the treatment of the whole breast (50 Gy). Chromosome aberrations in peripheral blood lymphocytes were measured using chemical-induced premature chromosome condensation combined with fluorescence in situ hybridization. RESULTS: Radiation treatment produced a significant increase in the yield of chromosomal aberrations. A large interindividual variability was observed. The variability was not related to field size, previous chemotherapy, or treatment morbidity. Chromosome aberrations in lymphocytes at the end of the treatment were significantly higher in the group of patients with no lymph nodes surgically removed before the treatment than in the group of patients with more than 10 lymph nodes removed. CONCLUSION: The number of lymph nodes within the radiation field is an important factor affecting the yield of radiation-induced chromosomal aberrations in breast cancer patients. PMID- 14529779 TI - Prognostic factors for disease-specific survival after first relapse of soft tissue sarcoma: analysis of 402 patients with disease relapse after initial conservative surgery and radiotherapy. AB - PURPOSE: To document the prognostic factors for survival of patients with soft tissue sarcoma sustaining a first relapse after definitive treatment. METHODS AND MATERIALS: The clinicopathologic features, relapse patterns, and disease-specific survival rates for 402 consecutive patients sustaining a first relapse of sarcoma after combined surgery and radiotherapy were retrospectively reviewed. Factors affecting disease-specific survival after relapse were evaluated with univariate and multivariate techniques. RESULTS: The median follow-up after relapse was 6.8 years. The overall disease-specific survival rate was 25%, 19%, and 16% at 5, 10, and 15 years, respectively, after the first relapse. The median survival duration was 21 months. Patients with an isolated local recurrence had a 5- and 10-year disease-specific survival rate of 48% and 46%, respectively, and those with an initial metastatic relapse had a disease-specific survival rate of 16% and 10%, respectively (p < 0.001). For isolated local recurrences, the independent determinants of survival were (favorable feature first) the primary tumor site (extremity and superficial trunk vs. head and neck and deep trunk); tumor grade (low and intermediate vs. high); time to recurrence (>12 vs. 5 cm). Although the development of subsequent metastasis was the major cause of death, a significant fraction of patients died of uncontrolled primary tumor. For patients presenting with metastasis as the first relapse, the time to metastasis was the major determinant of survival (>12 vs. < or =12 months). Long-term salvage was largely confined to patients who could and did undergo resection of relapsed disease, either local or metastatic. CONCLUSION: On the whole, patients whose sarcoma relapses fare poorly. However, select subgroups are potentially salvageable. Patients with an isolated local recurrence at sites other than the head and neck and deep trunk have a reasonable prospect for satisfactory outcome. Surgical resection of recurrences and metastases appears to play a major role in potential salvage. PMID- 14529780 TI - Stage III follicular lymphoma: long-term follow-up and patterns of failure. AB - PURPOSE: To analyze the long-term outcomes and pattern of failures for Stage III follicular lymphomas. METHODS AND MATERIALS: A retrospective review of all patients with Stage III follicular lymphoma presented to our institution between 1978 and 1993 was performed. One hundred ten patients were eligible and form the basis of this analysis. Fifty-seven patients were male. The median age was 57 years (range: 21-82 years). The treatments were as follows: chemotherapy alone (CTX), 39 patients; combined modality with chemotherapy and radiation therapy (CMT), 69; radiation therapy alone, 2. Radiation therapy fields were as follows: regional, 13 patients; extended, 6; subtotal, 44; and central lymphatic, 8. The median number of chemotherapy cycles was 8, and 98 patients received doxorubicin containing regimens. Enough information was available to calculate the International Prognostic Index for 107 patients. The following prognostic factors were examined for predictive value in overall survival (OS) and freedom from progression (FFP) by univariate and multivariate analyses: International Prognostic Index, gender, lactate dehydrogenase (LDH) (normal vs. elevated), B symptoms, performance status, beta-2 microglobulin, presence or absence of a bulky disease, age (60 years), number of sites of involvement, treatment (CTX vs. CMT), and pathology. To minimize patient selection biases given the nature of the retrospective analysis, the patterns of relapse were analyzed only for the patients who achieved a complete response. RESULTS: The median follow-up for the alive patients was 9.5 years (range: 1.1-19.5 years). Complete response was achieved in 80 patients: 24 of 39 patients in the CTX group (62%), 54 of 69 patients in the CMT group (78%), and 2 of 2 patients in the radiation therapy alone group. The actuarial 5- and 10-year OS rates were 65% and 42%, respectively. The 5- and 10-year FFP was 42% and 26%, respectively. Significant prognostic factors by multivariate analyses were age and LDH for OS and LDH for FFP. For complete responders, 5-year freedom from recurrence in the original sites of involvement (with or without recurrence in the new sites) was 43% for CTX and 60% for CMT patients (p = 0.03, Wilcoxon). Five-year freedom from isolated recurrence in the original sites of involvement was 60% for CTX and 69% for CMT patients (p = 0.17). The 5-year overall FFP was 43% for CTX patients and 56% for CMT patients (p = 0.06). CONCLUSION: Combined modality treatment seems to give an advantage in terms of disease control in the primary sites compared to chemotherapy alone, though the advantages in OS and FFP were not statistically significant in our patient population. By multivariate analyses, LDH was a significant prognostic indicator for OS and FFP, whereas age was for OS. PMID- 14529782 TI - Operations research applied to radiotherapy, an NCI-NSF-sponsored workshop February 7-9, 2002. PMID- 14529781 TI - The treatment of adults with medulloblastoma: a prospective study. AB - PURPOSE: To assess in a prospective trial the value of prognostic factors and the outcome of medulloblastoma in adults. METHODS AND MATERIALS: Patients (> or =18 years) with a histologic diagnosis of medulloblastoma were staged according to Chang et al.'s classification (low risk: T1, T2, T3a, M0, and no residual disease after surgery; high risk: T3b-T4, any M+ or postoperative presence of residual tumor). In low-risk patients, treatment consisted of 36 Gy to the craniospinal axis, supplemented by a local tumor dose of 18.8 Gy (total dose of 54.8 Gy). In high-risk patients, 2 cycles of "up-front chemotherapy" were delivered before the same radiation therapy, followed by maintenance chemotherapy if M1, M2, or M3 disease was present. RESULTS: Over a 12-year period, 36 evaluable patients were enrolled. Progression-free survival (PFS) at 5 years was higher in low-risk patients compared to the high-risk group: 76% +/- 14% (95% confidence interval [CI] = 52%-100%) vs. 61% +/- 11% (95% CI = 42%-87%). Patients with M- disease showed a significantly better outcome than M+ patients, with 75% showing PFS at 5 years vs. 45% (p = 0.01). CONCLUSION: The overall PFS observed is comparable to that obtained in pediatric series and suggests that a more effective therapy must be developed for high-risk patients. PMID- 14529783 TI - Intrinsic susceptibility to radiation-induced apoptosis of human lymphocyte subpopulations. AB - PURPOSE: With the aim to evaluate intrinsic radiosensitivity, the susceptibility of lymphocyte subpopulations to radiation-induced apoptosis was determined. The investigated parameters included measurement reliability, phenotypic variance, intra- and inter-individual variability, and correlations between radiation induced and spontaneous apoptosis. METHODS AND MATERIALS: Quiescent lymphocytes of 63 healthy volunteers, sampled up to four times over a 1-year period were gamma-irradiated in vitro. Subsequent apoptosis (annexin V) was measured for T4-, T8-, and B-lymphocyte subpopulations using 6-color flow cytometry. Spontaneous apoptosis was measured and radiosensitivity was quantified from the dose-effect curves. RESULTS: After thawing and short-term culture, both spontaneous apoptosis as well as radiation-induced apoptosis (radiosensitivity) differed among the three lymphocyte subpopulations, with T4 being most resistant, and B most sensitive. Spontaneous and radiation-induced apoptosis were correlated in all cell types, and variance between individuals was considerably higher than variance within individuals for both. A small but highly significant increase of both spontaneous and radiation-induced apoptosis was observed with age for T8, but not for T4 and B. Radiosensitivity of T8 and B proved to be sex-independent, whereas female T4 lymphocytes were less radiosensitive than those from males. T4 and T8 radiosensitivities were loosely correlated, and neither of them was related to B radiosensitivity. CONCLUSION: Tendency to spontaneous and radiation induced apoptosis of lymphocyte subpopulations differs among individuals. In addition, depending on the cell types, age and sex are factors influencing these parameters. PMID- 14529784 TI - Lipid a radiosensitizes hypoxic EMT-6 tumor cells: role of the NF-kappaB signaling pathway. AB - PURPOSE: Lipid A has shown promising immunostimulatory effects in both experimental tumor models and advanced stage cancer patients. This study examines whether lipid A may directly modulate the radioresponse of tumor cells by activating inducible nitric oxide synthase (iNOS) or cyclooxygenase-2 (COX-2) through nuclear factor-kappaB (NF-kappaB) signaling. METHODS AND MATERIALS: Hypoxic EMT-6 tumor cells were exposed to lipid A and analyzed for the level of COX-2 and iNOS by Western blotting and enzymatic assays. The hypoxic radioresponse of EMT-6 cells was estimated by clonogenic survival. The activation of NF-kappaB was examined by immunostaining of its p65 subunit and by luciferase reporter gene assay. RESULTS: Lipid A dose-dependently increased the expression and activity of iNOS with a maximal effect at plasma achievable concentrations of 3-30 micro g/mL. The COX-2 mediated production of prostaglandin E2 was constitutively high and further upregulated by lipid A. The radiosensitivity of hypoxic EMT-6 cells was increased up to 2.5 times and counteracted by the iNOS inhibitor aminoguanidine but not by the COX-2 inhibitor NS-398. The mechanism of radiosensitization was linked to NF-kappaB signaling, because its inhibition by phenylarsine oxide impaired both iNOS activation and radioresponse. CONCLUSION: Lipid A is an efficient hypoxic cell radiosensitizer at plasma relevant concentrations, which provides a rationale to combine lipid A with radiotherapy in further studies. PMID- 14529785 TI - Response to motexafin gadolinium and ionizing radiation of experimental rat prostate and lung tumors. AB - PURPOSE: To investigate the responses of two experimental rat tumors to single and fractionated X-ray doses whether or not combined with Motexafin gadolinium (MGd), and the distribution of MGd in R3327-MATLyLu (MLL) tumors using MRI. METHODS: L44 lung tumor in BN rats and MLL prostate tumor in Copenhagen rats were grown subcutaneously. MGd at concentrations of 8.7 to 25.1 micro mol/kg was administered 2 h before or just before treatments with single and fractionated X ray doses. Tumor volume growth delay was the endpoint used. The two-dimensional distribution of the MGd concentration in time was analyzed simultaneously in slices through the center of MLL tumors using MRI. Directly after the MRI experiments, tumor sections were stained for cytoplasm, nuclei, and microvessel endothelium. RESULTS: MGd at different concentrations administered a few minutes or 2 h before X-ray doses produced no radiation enhancement in the two tumor models. The MGd concentration as determined by MRI was maximal 5 min after injection and decreased slowly thereafter. In a representative section at the center of the MLL tumor, the microvessel density is nearly homogeneous and correlates with a nearly homogeneous MGd distribution. Hardly any MGd is taken up in underlying muscle tissue. CONCLUSION: No radiosensitization was observed for the different irradiation regimens. The distribution of MGd is nearly homogeneous in the MLL tumor and hardly any MGd is taken up in underlying muscles. Our negative results on radiosensitivity in our two tumor models raise questions about the efficacy of MGd as a general radiosensitizing agent. PMID- 14529786 TI - Subcutaneous administration of amifostine (ethyol) is equivalent to intravenous administration in a rat mucositis model. AB - PURPOSE: Amifostine (Ethyol) is currently approved for intravenous (IV) administration to prevent xerostomia in patients receiving radiotherapy for head and-neck cancer. Recently, subcutaneous (SC) administration has been explored as an alternative route. To determine whether SC administration was equivalent to IV administration, we used models to follow pharmacokinetics and oral mucosal protection in rats. METHODS: Amifostine was administered to rats at doses of 200, 100, or 50 mg/kg (1300, 650, or 325 mg/m(2)) IV or SC at various times before radiation at 15.3 Gy (protection studies) or harvest of blood and tissues for analysis by HPLC (pharmacokinetic studies). RESULTS: Amifostine administered IV or SC 1 h before radiation protected rats from mucositis, but the protective effect was more prolonged when amifostine was administered SC. Tissue levels of the active metabolite (WR-1065) were equivalent after SC administration. The correlation between tissue levels of WR-1065 and protection was strong, but that between blood levels of WR-1065 and protection was only weak. CONCLUSION: These data demonstrate that, in a rat model, SC administration of amifostine was at least as effective as that by IV. PMID- 14529787 TI - Improved free vascular graft survival in an irradiated surgical site following topical application of rVEGF. AB - PURPOSE: Wound healing disorders following surgery in preirradiated tissue are clinically well known and may even become more crucial with the increasing use of neoadjuvant chemoradiation protocols. Both the expression of vascular endothelial growth factor (VEGF) and endoglin (CD105) play a key role in neovascularization and wound healing after soft tissue grafts in irradiated and nonirradiated tissue. Modulation of neovascularization through the application of recombinant VEGF (rVEGF) may be a therapeutic option to reduce wound healing disorders in irradiated tissue. An experimental in vivo model was used to study the possible role of rVEGF for reduction of wound healing disorders and the promotion of neovascularization. METHODS AND MATERIALS: A free myocutaneous gracilis flap was transplanted from the groin into the neck region of Wistar rats (weight 300-500 g) with and without previous irradiation of the neck region with 40 Gy: Group 1 (n = 7) radiotherapy alone; Group 2 (n = 14) flap transplantation alone and rVEGF; Group 3 (n = 14) radiotherapy, transplantation, and rVEGF. Time interval between irradiation and grafting was 10 +/- 1 day. 1.0 micro g rVEGF/500 microL phosphate-buffered saline was applied s.c. intraoperatively and on Days 1 through 7. Neovascularization (CD105) and endogenous VEGF expression were analyzed by means of immunohistochemistry on Days 3, 5, 7, 14, and 28 postoperatively and quantified as labeling indices (LI). RESULTS: After irradiation there was a continuous significant reduction of the cytoplasmic VEGF expression (MEAN LI: 0.018 +/- 0.048) compared with the nonirradiated control (mean LI: 0.042 +/- 0.006) (p < 0.001). VEGF expression after flap transplantation without irradiation after VEGF application was at a constantly higher level from Day 3 (mean LI: 0.044 +/- 0.01) to Day 28 postoperatively compared with the control group (Day 3, mean LI: 0.028 +/- 0.006) (p < 0.001). As an indication of increased neovascularization after the local application of rVEGF, a significantly increased expression of CD105 was found in the transition area and graft bed from Day 7 on (p < 0.001). After irradiation and grafting there was a significant overall increase in the VEGF- and CD105-expression throughout Day 28 after rVEGF in the transition area (p < 0.001). CONCLUSION: Whereas irradiation alone led to a downregulation of the endogenous VEGF expression, rVEGF application resulted in an increased expression and in a CD105 associated neovascularization after soft tissue grafting in irradiated tissues. Application of rVEGF may enable modulation of wound healing by influencing neovascularization. This could indicate a possible clinical approach for reducing fibrosis and chronic wound healing disorders in irradiated tissues. PMID- 14529788 TI - Cytotoxicity of perillyl alcohol against cancer cells is potentiated by hyperthermia. AB - PURPOSE: Perillyl alcohol (POH) (4-isopropenyl-cyclohexenecarbinol) is a member of the monoterpenes, which are present in various fruits and vegetables. POH has been demonstrated to be cytotoxic against a variety of experimental cancer cells in vitro and in vivo. Phase I clinical trials have indicated that POH may be useful for human tumor treatment. The purpose of our study was to reveal whether the anticancer effect of POH could be enhanced by hyperthermia. METHODS AND MATERIALS: SCK mammary carcinoma cells of A/J mice were used. The effects of POH or hyperthermia alone were studied by incubating the cells during exponential growth phase in culture with 0.25-1.0 mM of POH at 37 degrees C for varying lengths of time or heating cells at 41-43 degrees C for varying lengths of time. The combined effect of POH and hyperthermia was investigated by heating the cells with 1 mM of POH at 41-43 degrees C for varying lengths of time. The effects of the treatments were evaluated using the clonogenic cell survival assay and three types of apoptosis assays. RESULTS: An incubation of SCK cells with 1 mM of POH at 37 degrees C for 60 min or hyperthermia at 43 degrees C for 1 h decreased clonogenic cell survival to 40% and 60%, respectively. When the cells were heated at 43 degrees C for 1 h in the presence of 1 mM of POH, clonogenic cell survival decreased to 0.2%, indicating that hyperthermia potentiated the effect of POH to cause clonogenic cell death. Hyperthermia also markedly increased the degree of POH-induced apoptosis. CONCLUSION: Hyperthermia synergistically potentiates the cytotoxicity of naturally occurring POH against cancer cells. PMID- 14529789 TI - Heat shock protein 70 (Hsp70) membrane expression on head-and-neck cancer biopsy a target for natural killer (NK) cells. AB - PURPOSE: Heat shock protein 70 (Hsp70) was detected on the cell membrane of human tumor cell lines, but not on normal cells. Here we studied Hsp70 membrane expression as a target for natural killer (NK) cells on tumor material and control tissues of head-and-neck cancer patients. METHODS AND MATERIALS: Membrane bound Hsp70 was determined by flow cytometry on single-cell suspensions of tumors and the corresponding normal tissues of head-and-neck cancer patients. The cytolytic activity of NK cells against Hsp70-positive tumor cells was measured in a standard cytotoxicity assay. RESULTS: In total, 54 of 74 primary tumors were found to be Hsp70 membrane-positive (73%); tongue/mouth, 21 of 24 (88%); oropharynx, 13 of 20 (65%); hypopharynx, 3 of 6 (50%); larynx, 8 of 11 (73%); trachea 1 of 2 (50%); esophagus, 4 of 5 (80%); lymph node metastases, 4 of 6 (67%). The corresponding control tissue was negative for membrane-bound Hsp70. Biopsies (6 of 6) of patients after in vivo gamma-irradiation (fractionated 5 x 2 Gy) were strongly Hsp70 membrane-positive. Irradiated, Hsp70-positive tumor cells are targets for Hsp70-peptide stimulated NK cells. CONCLUSION: An irradiation inducible, tumor-selective Hsp70 membrane localization provides a target structure for Hsp70-peptide stimulated human NK cells. PMID- 14529790 TI - The linear-quadratic model and fractionated stereotactic radiotherapy. AB - PURPOSE: To determine the dose per fraction that could be used when gamma knife or linear accelerator-based stereotactic treatments are delivered in 2 or more fractions. METHODS AND MATERIALS: The linear-quadratic (LQ) model was used to calculate the dose per fraction for a multiple-fraction regimen which is biologically equivalent to a given single-fraction treatment. The results are summarized in lookup tables. RESULTS AND CONCLUSION: The tables can be used by practicing clinicians as a guide in planning fractionated treatment. For the large doses used in typical stereotactic treatments and for small fraction numbers, the model is not very sensitive to the value of the alpha/beta ratio in the LQ model. A simple rule of thumb is found that for two-fraction and three fraction treatments the dose per fraction is roughly two-thirds and one-half of the single-fraction treatment dose, respectively. PMID- 14529791 TI - The impact of geometric uncertainty on hypofractionated external beam radiation therapy of prostate cancer. AB - PURPOSE: Recent publications indicate alpha/beta for prostate carcinoma could be lower than assumed. Therefore, hypofractionation might increase the therapeutic ratio. However, patient repositioning and organ motion may affect hypofractionated treatments more than conventional treatments. Our purpose is to evaluate the potential impact of geometric uncertainties on hypofractionated treatments. METHODS AND MATERIALS: Tumor control probability (TCP) and normal tissue complication probability (NTCP) are calculated for simulated conventional and hypofractionated treatments, assuming alpha/beta of 1.5 Gy for prostate and 3.0 Gy for rectum. A Monte Carlo simulation randomly samples systematic and random displacements and produces the cumulative dose distribution for the prostate and rectum. The limiting number of fractions and the impact of different alpha/beta values are also explored. RESULTS: A consistent but small reduction in TCP is seen with hypofractionation (generally <1%) as a result of geometric uncertainties. Escalated hypofractionation seems to allow large TCP gains ( approximately 20%) without increasing NTCP. Treatments of five fractions seem to affect outcome minimally. The alpha/beta value has a much greater impact on TCP than geometric uncertainties. CONCLUSION: The potential increased influence of geometric uncertainties on hypofractionation seems small. Limited knowledge of radiobiologic response is likely a greater obstacle to prostate hypofractionation than geometric uncertainties. PMID- 14529792 TI - A method for increased dose conformity and segment reduction for SMLC delivered IMRT treatment of the prostate. AB - PURPOSE: The focus of this work is to develop a practical planning method that results in increased dose conformity and reduced treatment time for segmental multileaf collimation (sMLC) based intensity-modulated radiation therapy (IMRT) delivery. METHODS AND MATERIALS: Additional regions for dose constraint are introduced within the normal tissue during the planning process by designing a series of concentric ellipsoids around the target. A dose gradient is then defined by assigning dose constraints to each concentric region. The technique was tested at two centers and data for 26 and 10 patients, respectively, are presented allowing for differences in treatment technique, beam energy, ellipsoid definition, and prescription dose. At both centers, a series of patients previously treated for prostate cancer with IMRT were selected, and comparisons were made between the original and new plans. RESULTS: While meeting target dose specifications and normal tissue constraints, the average number of beam directions decreased by 1.6 with a standard error (SE) of 0.1. The average time for delivery at center 1 decreased by 29.0% with an SE of 2.0%, decreasing from 17.5 min to 12.3 min. The average time for delivery at center 2 decreased by 29.9% with an SE of 3.8%, decreasing from 11 min to 7.7 min. The amount of nontarget tissue receiving D(100) decreased by 15.7% with an SE of 2.4%. Nontarget tissue receiving D(95), D(90), and D(50) decreased by 16.3, 15.1, and 19.5%, respectively, with SE values of approximately 2% at center 1. Corresponding values for D(100), D(95), D(90), and D(50) decreased by 13.5, 16.7, 17.1, and 5.1%, respectively, with SE values of less than 3% at center 2. CONCLUSION: By designating subsets of tissue as concentric regions around the target(s) and carefully defining each region's dose constraints, we have gained an increased measure of control over the region outside the target boundaries. This increased control manifests as two distinct endpoints that are beneficial to the IMRT process: increased dose conformity and decreased treatment time. PMID- 14529793 TI - Radiation treatment planning with an integrated positron emission and computer tomography (PET/CT): a feasibility study. AB - PURPOSE: To investigate the usefulness of hardware coregistered PET/CT images for target volume definition. METHODS AND MATERIALS: Thirty-nine patients presenting with various solid tumors were investigated. CT and a FDG-PET were obtained in treatment position in an integrated PET/CT scanner, and coregistered images were used for treatment planning. First, volume delineation was performed on the CT data. In a second step, the corresponding PET data were used as an overlay to the CT data to define the target volume. Delineation was done independently by two investigators. RESULTS: Coregistered PET/CT showed good fusion accuracy. The GTV increased by 25% or more because of PET in 17% of cases with head-and-neck (2/12) and lung cancer (1/6), and in 33% (7/21) in cancer of the pelvis. The GTV was reduced > or =25% in 33% of patients with head-and-neck cancer (4/12), in 67% with lung cancer (4/6), and 19% with cancer of the pelvis (4/21). Overall, in 56% (22/39) of cases, GTV delineation was changed significantly if information from metabolic imaging was used in the planning process. The modification of the GTV translated into altered PTV changes exceeding >20% in 46% (18/39) of cases. With PET, volume delineation variability between two independent oncologists decreased from a mean volume difference of 25.7 cm(3) to 9.2 cm(3) associated with a reduction of the standard deviation from 38.3 cm(3) to 13.3 cm(3) (p = 0.02). In 16% of cases, PET/CT revealed distant metastasies, changing the treatment strategy from curative to palliative. CONCLUSION: Integrated PET/CT for treatment planning for three-dimensional conformal radiation therapy improves the standardization of volume delineation compared with that of CT alone. PET/CT has the potential for reducing the risk for geographic misses, to minimize the dose of ionizing radiation applied to non-target organs, and to change the current practice to three-dimensional conformal radiation therapy planning by taking into account the metabolic and biologic features of cancer. The impact on treatment outcome remains to be demonstrated. PMID- 14529794 TI - Validation of active breathing control in patients with non-small-cell lung cancer to be treated with CHARTWEL. AB - PURPOSE: Active breathing control (ABC) was validated using patients with non small-cell lung cancer (NSCLC) to be treated with continuous hyperfractionated accelerated radiotherapy weekend-less (CHARTWEL). Effects of breath hold (BH) on accuracy and normal tissue doses were evaluated. METHODS AND MATERIALS: Eleven patients were studied. Immediately after a free breathing (FB) planning scan, two ABC scans (ABC 1 and 2) were performed to assess intrafraction variation. A third ABC scan (ABC 3) was performed some weeks later to assess interfraction variation. Assisted BH was set at 75% of vital capacity and reproducibility assessed using computed tomography (CT) lung volumes. Planning target volumes (PTVs), doses to lung and spinal cord for FB and ABC 1 scans were compared. RESULTS: Results were available for 10 patients. Disease and elective nodal regions were easier to define on ABC scans making PTVs smaller. ABC lung volumes showed no significant variation over several weeks, percentage volume of whole lung receiving > or =20 Gy (V(20)) was reduced in all (median 6.4%, p = 0.005), and spinal cord dose in 80% (median 1.03 Gy, p = 0.02), of the plans. CONCLUSION: ABC allowed reproducible BH, and enabled better delineation of tumor and normal structures, as well as reduction in PTV, V(20), and spinal cord dose. PMID- 14529795 TI - Potential for reduced toxicity and dose escalation in the treatment of inoperable non-small-cell lung cancer: a comparison of intensity-modulated radiation therapy (IMRT), 3D conformal radiation, and elective nodal irradiation. AB - PURPOSE: To systematically evaluate four different techniques of radiation therapy (RT) used to treat non-small-cell lung cancer and to determine their efficacy in meeting multiple normal-tissue constraints while maximizing tumor coverage and achieving dose escalation. METHODS AND MATERIALS: Treatment planning was performed for 18 patients with Stage I to IIIB inoperable non-small-cell lung cancer using four different RT techniques to treat the primary lung tumor +/- the hilar/mediastinal lymph nodes: (1) Intensity-modulated radiation therapy (IMRT), (2) Optimized three-dimensional conformal RT (3D-CRT) using multiple beam angles, (3) Limited 3D-CRT using only 2 to 3 beams, and (4) Traditional RT using elective nodal irradiation (ENI) to treat the mediastinum. All patients underwent virtual simulation, including a CT scan and (18)fluorodeoxyglucose positron emission tomography scan, fused to the CT to create a composite tumor volume. For IMRT and 3D-CRT, the target included the primary tumor and regional nodes either > or =1.0 cm in short-axis dimension on CT or with increased uptake on PET. For ENI, the target included the primary tumor plus the ipsilateral hilum and mediastinum from the inferior head of the clavicle to at least 5.0 cm below the carina. The goal was to deliver 70 Gy to > or =99% of the planning target volume (PTV) in 35 daily fractions (46 Gy to electively treated mediastinum) while meeting multiple normal tissue dose constraints. Heterogeneity correction was applied to all dose calculations (maximum allowable heterogeneity within PTV 30%). Pulmonary and esophageal constraints were as follows: lung V(20) < or =25%, mean lung dose < or =15 Gy, esophagus V(50) < or =25%, mean esophageal dose < or =25 Gy. At the completion of all planning, the four techniques were contrasted for their ability to achieve the set dose constraints and deliver tumoricidal RT doses. RESULTS: Requiring a minimum dose of 70 Gy within the PTV, we found that IMRT was associated with a greater degree of heterogeneity within the target and, correspondingly, higher mean doses and tumor control probabilities (TCPs), 7%-8% greater than 3D-CRT and 14%-16% greater than ENI. Comparing the treatment techniques in this manner, we found only minor differences between 3D-CRT and IMRT, but clearly greater risks of pulmonary and esophageal toxicity with ENI. The mean lung V(20) was 36% with ENI vs. 23%-25% with the three other techniques, whereas the average mean lung dose was approximately 21.5 Gy (ENI) vs. 15.5 Gy (others). Similarly, the mean esophagus V(50) was doubled with ENI, to 34% rather than 15%-18%. To account for differences in heterogeneity, we also compared the techniques giving each plan a tumor control probability equivalent to that of the optimized 3D-CRT plan delivering 70 Gy. Using this method, IMRT and 3D-CRT offered similar results in node-negative cases (mean lung and esophageal normal tissue complication probability [NTCP] of approximately 10% and 2%-7%, respectively), but ENI was distinctly worse (mean NTCPs of 29% and 20%). In node positive cases, however, IMRT reduced the lung V(20) and mean dose by approximately 15% and lung NTCP by 30%, compared to 3D-CRT. Compared to ENI, the reductions were 50% and >100%. Again, for node-positive cases, especially where the gross tumor volume was close to the esophagus, IMRT reduced the mean esophagus V(50) by 40% (vs. 3D-CRT) to 145% (vs. ENI). The esophageal NTCP was at least doubled converting from IMRT to 3D-CRT and tripled converting from IMRT to ENI. Finally, the total number of fractions for each plan was increased or decreased until all outlined normal-tissue constraints were reached/satisfied. While meeting all constraints, IMRT or 3D-CRT increased the deliverable dose in node-negative patients by >200% over ENI. In node-positive patients, IMRT increased the deliverable dose 25%-30% over 3D-CRT and 130%-140% over ENI. The use of 3D-CRT without IMRT increased the deliverable RT dose >80% over ENI. Using a limited number of 3D-CRT beams decreased the lung V(20), mean dose, and NTCP in node-positive patients. CONCLUSION: The use of 3D-CRT, particul mean dose, and NTCP in node-positive patients. The use of 3D-CRT, particularly with only 3 to 4 beam angles, has the ability to reduce normal-tissue toxicity, but has limited potential for dose escalation beyond the current standard in node-positive patients. IMRT is of limited additional value (compared to 3D-CRT) in node negative cases, but is beneficial in node-positive cases and in cases with target volumes close to the esophagus. When meeting all normal-tissue constraints in node-positive patients, IMRT can deliver RT doses 25%-30% greater than 3D-CRT and 130%-140% greater than ENI. Whereas the possibility of dose escalation is severely limited with ENI, the potential for pulmonary and esophageal toxicity is clearly increased. PMID- 14529797 TI - Intracavitary balloon brachytherapy for breast cancer: surgical considerations. in regard to Keisch M, Vicini F, Kuske RR, et al., IJROBP 2003;55:289-293. PMID- 14529796 TI - Three-dimensional photon dosimetry: a comparison of treatment of the intact breast in the supine and prone position. AB - PURPOSE: To compare the adequacy of target coverage, dose homogeneity, and volume of normal tissue irradiated in treatment of the intact breast in the supine and prone position. METHODS AND MATERIALS: Fifteen patients with early breast cancer who presented for treatment to the intact breast after excisional biopsy were studied. A specially designed device was used for the prone setup to displace the contralateral breast away from the tangential field borders. Treatment planning computed tomography was performed for each patient in both the supine and prone positions. Dosimetric data were obtained in both positions and isodose distributions were calculated for each patient in both positions. RESULTS: The volume of breast receiving greater than 5% of the prescribed dose was significantly less in the prone position. Medial wedges were either not used or their angles were reduced for all patients in the prone position compared with the supine position. The average volume of lung receiving >10 Gy and >20 Gy was significantly less in the prone positions. The volume of heart irradiated at critical dose levels did not vary consistently in the prone and supine positions. The integral dose delivered to the contralateral breast was not significantly different. CONCLUSION: Treatment of the intact breast in the prone position may result in improved dose homogeneity within the target volume as well as sparing of normal lung compared with treatment in the conventional supine position. PMID- 14529799 TI - Drinking decreases the noradrenaline release in the median preoptic area caused by hypovolemia in the rat. AB - Previous observations have suggested that the noradrenergic system in the median preoptic nucleus (MnPO) is implicated in the regulation of body fluid balance and cardiovascular function. The present study was carried out to investigate whether water intake alters the release of noradrenaline (NA) in the MnPO area caused by hypovolemia in freely moving rats. Nonhypotensive hypovolemia was induced by subcutaneous polyethylene glycol (PEG), and extracellular levels of NA were measured using intracerebral microdialysis techniques. Subcutaneous injections of PEG (30%, 5 ml) significantly enhanced the NA release in the MnPO area. Water ingestion significantly attenuated the elevation in the NA release in the MnPO area induced by the PEG treatment. These results show the involvement of the noradrenergic system in the MnPO in the maintenance of body fluid volume, and suggest that the system may play an important role in the elicitation of hypovolemia-induced dipsogenic response. PMID- 14529800 TI - Effects of methylphenidate, desipramine, and L-dopa on attention and inhibition in children with Attention Deficit Hyperactivity Disorder. AB - The objective of this study was to investigate the effects of methylphenidate (MPH) on attention and inhibition in children with Attention Deficit Hyperactivity Disorder (ADHD) and to establish what the relative contributions of the noradrenergic and dopaminergic systems to this effect were. In addition to MPH, two other drugs were administered in order to affect both transmitter systems more selectively, L-dopa (dopamine (DA) agonist) and desipramine (DMI) (noradrenaline (NA) re-uptake inhibitor). Sixteen children with ADHD performed a stop-task, a laboratory task that measures the ability to inhibit an ongoing action, in a double-blind randomized within-subjects design. Each child received an acute clinical dose of MPH, DMI, L-dopa, and placebo; measures of performance and plasma were determined. The results indicated that inhibition performance was improved under DMI but not under MPH or L-dopa. The response-time to the stop signal was marginally shortened after intake of DMI. MPH decreased omission and choice-errors and caused faster reaction times to the trials without the stop tone. No effects of L-dopa whatsoever were noted. Prolactin levels were increased and 5-HIAA levels were lowered under DMI relative to placebo. It is suggested that the effects of MPH on attention are due to a combination of noradrenergic and dopaminergic mechanisms. The improved inhibition under DMI could be serotonergically mediated. PMID- 14529801 TI - Rat behavior in two models of anxiety and brain [3H]muscimol binding: pharmacological, correlation, and multifactor analysis. AB - The contribution of GABAergic mechanisms to rat emotional behavior in two animal models of anxiety (open field test of neophobia and aversively conditioned freezing reaction), was confirmed by pharmacological analysis, using anxiolytic (midazolam) and anxiogenic (picrotoxin) compounds. Both substances are known to modulate GABA(A) receptors' activity in a positive or negative manner, respectively. It seemed, therefore, worthwhile to check whether the behavioral parameters measured in these animal models of anxiety correlate with [3H]muscimol binding (a highly selective GABA(A) receptor ligand) in different brain structures of nai;ve rats, with a view to establish the role of genetically determined expression of local GABA(A) receptors in the organization of rat emotional and motor behavior. Correlation analysis revealed no links between individually determined expression of GABA(A) receptors (quantitative receptor autoradiography) in the brain structures, and the emotional behavior of nai;ve, drug-free animals, in both tests. Factor analysis confirmed that animal behavior in both tests was under control of different central processes. Moreover, none of the behavioral and ligand binding parameters loaded on the same factor, confirming the negative results of the correlation study. The present results indicate that the origin of emotions is a complex phenomenon, probably involving the interaction between GABA-ergic innervation of many brain structures. PMID- 14529802 TI - The emotional reactivity increase at mid-pregnancy is attenuated in female rats handled during their infancy. AB - In rats, while pregnancy has been reported to increase emotional reactivity, postnatal handling, consisting in a short daily maternal separation during early life, has been shown to reduce it in several behavioural tests. In the present study, we investigated the potential effects of handling in pregnant rats confronted to various stressful situations. For that purpose, groups of DA/Han female rats were tested either in the elevated plus maze test, in the probe burying test or in a cat exposure test. At the age of 10-14 weeks, control and handled females were mated. Pregnant females were isolated (day 0 of gestation) as well as some non-pregnant females (day 0 of isolation). On day 12, pregnant and non-pregnant females were subjected to one of the three emotional reactivity tests. Our results show that pregnant females (control and handled) were more reactive than non-pregnant ones in two of the three testing situations, namely the elevated plus maze and the probe burying test. In particular, pregnant females spent less time in the open arms of the elevated plus maze than their non pregnant counterparts and their number of close sniffing towards the probe was decreased in the burying test. Comparatively, beneficial effects of postnatal handling were obvious in the three experiments, both for non-pregnant and pregnant females. We conclude (1) that pregnancy increases emotional reactivity at day 12 of pregnancy, particularly in tests involving low or mildly stressful conditions, and (2) that postnatal handling can partially reverse those effects in all three tests, even in the more stressful situation represented by the cat exposure. PMID- 14529803 TI - Serotonin synthesis inhibition in olivo-cerebellar system attenuates harmaline induced tremor in Swiss albino mice. AB - Recent experimental evidences point to the active role of central serotonin (5 HT) elicited mechanisms in the pathogenesis of tremor. The present study was undertaken to investigate the effects of p-chlorophenylalanine (pCPA), a specific tryptophan hydroxylase inhibitor and a central 5-HT depletor, on the neurochemical processes that occur synchronously in olivary nucleus (ON) and cerebellum during harmaline-induced tremor in mice. Tremor appeared by 3-4 min following harmaline administration, and reached its peak by 25 min for the doses (10-30 mg/kg) studied. Peak of harmaline-tremor coincided with increases in 5-HT in ON and cerebellum, as assayed employing HPLC-electrochemistry. Administration of pCPA caused significant depletion in 5-HT level in both the regions analyzed, and also significantly inhibited harmaline-induced tremor. Our present results support the earlier electrophysiological evidences that harmaline-induced tremor originates from ON, and confirm the role of 5-HT in the genesis of this motor neuronal dysfunction. PMID- 14529804 TI - Vitamin A deficiency and relational memory deficit in adult mice: relationships with changes in brain retinoid signalling. AB - Vitamin A and its derivatives, the retinoids, have recently been reported to be implicated in the synaptic plasticity of the hippocampus and in cognitive functions. Acting via transcription factors, retinoids can regulate gene expression via their nuclear receptors [retinoic acid receptors (RARs) and retinoid X receptors (RXRs)]. We recently showed that a moderate (about 30%) hypoexpression of brain (and hippocampal) retinoid signalling, like that naturally occurring in the aged brain of mice, might be related to a selective relational memory deficit. To further assess this hypothesis, the present study investigated the effects of Vitamin A deprivation of varying duration both on the brain expression of retinoid receptors (RARbeta and RXRbeta/gamma) and two associated target genes [tissue-type transglutaminase (tTG) and neurogranin, (RC3)], and on radial maze discrimination learning using young adult mice as subjects. We observed that irrespective of its duration (i.e. 31 or 39 weeks), Vitamin A deprivation resulted in a significant reduction (25-30%) in the expression of brain RARbeta, RXRbeta/gamma and tTG mRNAs. Conversely, only the 39 week condition was found to induce a significant decrease in brain RC3 mRNAs contents and a selective relational memory impairment. Finally, daily administration of retinoic acid (RA) failed to reverse the 39-week Vitamin A deficiency (VAD)-related cognitive deficit and to fully normalise the associated brain retinoid hyposignalling. In particular, there was no evidence for an up regulating effect of RA on whole brain (and hippocampal) RC3 mRNAs of the 39-week depleted mice. The results show that post-natal VAD may induce a selective memory impairment and give further support to the hypothesis that the fine regulation of retinoid-mediated gene expression is important for optimal brain functioning and higher cognition. PMID- 14529805 TI - Head-only exposure to GSM 900-MHz electromagnetic fields does not alter rat's memory in spatial and non-spatial tasks. AB - Over the last decade, exposure to high frequency (2450 MHz) electromagnetic fields (EMFs) has been found to induce performance deficit in rodents in spatial memory tasks. As concern was expressed about potential biological effects of mobile communication microwaves, studies testing the effects of signals such as GSM were required. In a previous study, using head-only exposure to 900 MHz GSM EMF, we could not demonstrate any behavioural deficit in two simple learning tasks. The present study aimed at extending these results with more complex spatial learning tasks and a non-spatial task. In a first experiment, rats were trained in a radial-arm maze with a 10-s confinement between each visited arm. In a second experiment, a 15-min intra-trial delay was introduced after four visited arms. In a third experiment, non-spatial memory was tested in an object recognition task. In all experiments, performance of the head-only exposed rats (1 and 3.5 W/kg) was compared with that of sham and control rats. In the first experiment, a slightly improved performance was found after 3.5 W/kg exposure, a result that was not observed in the delay-task. In the third experiment, although some effects on exploratory activity were found, recognition memory was unaffected in exposed rats. Altogether, this set of experiments provides no evidence indicating that spatial and non-spatial memory can be affected by a 45 min head-only exposure to 900 MHz GSM EMF. PMID- 14529806 TI - Roles of the auditory midbrain and thalamus in selective phonotaxis in female gray treefrogs (Hyla versicolor). AB - Diencephalic and midbrain auditory nuclei are involved in the processing of auditory communication signals in anurans [Comparative Hearing: Fish and Amphibians, Springer-Verlag, New York, 1999, p. 218], but their exact roles in acoustically guided behavior, such as female phonotaxis, are unclear. To address this question, behavioral experiments were combined with lesions of dorsal thalamic nuclei and the midbrain torus semicircularis. Females were tested in two alternative-forced-choice phonotactic experiments before and after a defined brain area was lesioned. During phonotactic tests, females had to choose between a "standard" synthetic call and one of three different variants, each of which had a single acoustic property (pulse rate, pulse rise-time, sound spectrum) that differed from the standard synthetic call. Results showed that dorsomedial thalamus lesions produced little or no effect on phonotaxis. In contrast, superficial and deep thalamus lesions, as well as lesions of the torus semicircularis, significantly decreased the number of phonotactic responses and increased the response time. Superficial thalamus lesions also abolished or reversed preferences for the standard call in the rise-time and sound spectrum tests. This effect is likely to have been caused by an imbalance in the stimulation of the thalamus by the low- and high-frequency pathways because these preferences were not affected in animals with more extensive lesions that included the superficial thalamus. Our data suggest that the torus semicircularis, but not the dorsal thalamus is crucial for phonotaxis in gravid, reproductively active females. Although dorsal thalamic nuclei seem to play a role in spectral sensitivity, they may additionally have motivational or attentional functions that contribute to achieving a state of phonotactic readiness. PMID- 14529807 TI - Selective induction of c-Fos immunoreactivity in the prelimbic cortex during reinstatement of heroin seeking induced by acute food deprivation in rats. AB - We previously reported that acute 1-day food deprivation reinstates heroin seeking in rats. The goal of the present study was to begin identifying brain sites potentially involved in this effect. For this purpose, we measured, by immunohistochemistry, the expression of c-Fos following a test for food deprivation-induced reinstatement. Groups of rats (n=9-10 per group) were trained to lever-press for heroin (0.05-0.1mg/kg/infusion) or saline for 10 days (9 h/day); each infusion was paired with a cue light. Rats were then given 10 days of extinction during which the heroin and saline syringes were removed. Next, a test for reinstatement was conducted after exposure to 0 (baseline) or 1-day food deprivation. During training, lever pressing for heroin increased over days, while responding for saline infusions paired with the cue light decreased over time. During extinction, responding on the heroin-paired lever decreased over time, while responding on the saline-paired lever remained low. In heroin-trained rats, food deprivation induced a large increase in responding on the lever associated with drug infusions. Surprisingly, food deprivation also modestly increased responding in the saline-trained rats. Food deprivation selectively increased c-Fos immunoreactivity (IR) in the prelimbic cortex of heroin-trained, but not saline-trained, rats (n=4 per condition). Food deprivation also increased c-Fos IR in both heroin- and saline-trained rats in the basolateral amygdala and the ventrolateral bed nucleus of stria terminalis (BNST), but had no effect on c Fos expression in the dorsolateral BNST, cingulate cortex, nucleus accumbens, and central amygdala. These results raise the possibility that the prelimbic cortex is involved in food deprivation-induced reinstatement of heroin seeking. PMID- 14529808 TI - Generalisation of conditioned fear and its behavioural expression in mice. AB - Mice are favourite subjects in molecular and genetic memory research and frequently studied with classical fear conditioning paradigms that use an auditory cue (conditioned stimulus, CS(+)) to predict an aversive, unconditioned stimulus (US). Yet the conditions that control fear memory specificity and generalisation and their behavioural expression in such conditioned mice have not been analysed systematically. In the current study we addressed these issues in the most widely used mouse strain of behavioural genetics, C57Bl/6. In keeping with findings in other species we demonstrate the dependence of fear memory generalisation on training intensity (i.e. both US intensity and the number of CS(+) and US applied) after both excitatory (explicitly paired presentation of CS(+) and US) and inhibitory training (explicitly unpaired presentation of CS(+) and US). Furthermore, inhibitory overtraining was associated with changes of uncued anxiety-like behaviour in a light/dark exploration test, indicative of an emotional sensitisation reaction as consequence of a lack of US predictability. Together our results describe the qualitatively and quantitatively different increases of defensive behaviour in response to conditioned stimuli of different salience and identify training conditions that lead to fear memory generalisation and emotional sensitisation in C57Bl/6 inbred mice. PMID- 14529809 TI - The nucleus accumbens shell is critical for normal expression of pup-retrieval in postpartum female rats. AB - The nucleus accumbens (NA) plays an important modulatory role in the control of normal expression of maternal behavior (termed maternal performance). The present study investigated the relative functions of two subregions of the NA (the shell and core) in maternal performance. Electrolytic lesions of the shell or core were performed either before parturition or immediately after a varying amount of maternal experience (none, 1 or 24h) during the immediate postpartum period. Maternal performance was tested on Day 1 postpartum and re-tested 9 days later using a pup sensitization technique. Results show that lesions of the shell, but not the core, significantly disrupted pup-retrieval: the shell-lesioned rats took significantly longer to finish retrieving all test pups, but their retrieval latency for the first pup was not affected. Neither lesion affected other components of maternal behavior (pup licking, nest building and nursing). These findings suggest that the shell, but not the core, is critical for the normal expression of pup-retrieval behavior possibly through its role in maintaining maternal motivation or attention. PMID- 14529810 TI - The magnitude and the extinction duration of the cocaine-induced conditioned locomotion-activated response are related to the number of cocaine injections paired with the testing context in C57BL/6J mice. AB - Behavioural activation repeatedly induced by a stimulant in rodents can persist in the absence of the drug if the animals are tested in the context where the drug was previously given, a phenomenon often explained in terms of Pavlovian conditioning. The aim of this study was to verify whether the amplitude of the putative CR (the drug-like activity) increases with the number of the US-CS associations (the number of drug-context pairings), one of the most representative rules of Pavlovian conditioning. The effect of the number of trials on the speed of extinction was also considered. C57BL/6J mice received 3, 6 or 12 once-daily injections of either saline or 12 mg/kg (-)-cocaine hydrochloride (s.c.) in the same test context, a photocell activity-box in which they were tested for 60 min after every injection. Other groups received the same treatments outside of the test context (being placed in a novel cage tub after each injection). Twenty-four hours after the last treatment session, all mice were challenged with saline in the test context (test for conditioned activity), extinction sessions taking place on the three subsequent days. Sensitisation to the locomotor-activating effect of cocaine developed only amongst the animals injected 6 or 12 times, the magnitude of the last sensitised response being comparable for these two injections regimen. On saline challenge, only the animals that had received 6 or 12 cocaine injections showed significant conditioned activity (CR), with the greatest response occurring following 12 injections. The 6-trial group reached the level of non-significance after fewer extinction sessions than the 12-trial group; however, the rates of extinction did not differ (comparable regression coefficients and quasi-parallel curves). These results suggest that the amplitude of the CR (cocaine-like stimulation after saline), and perhaps less convincingly the duration of extinction, are functions of the number of the US-CS (cocaine-context) pairings, supporting the Pavlovian nature of post-sensitisation placebo drug-like effects. PMID- 14529811 TI - Stereotypies in caged parrots, schizophrenia and autism: evidence for a common mechanism. AB - Spontaneously occurring abnormal behaviors in animals have recently received considerable attention, both in veterinary medicine and as a potential model for abnormal behavior in several human mental disorders. Stereotypies are abnormal repetitive, unvarying, and functionless behaviors that are often performed by captive and domesticated animals housed in barren environments. They closely resemble the stereotypies of autistic and mentally retarded patients, stereotypies of unmedicated chronic schizophrenic patients, certain classes of simple tic in Tourette's syndrome, and several drug-induced behaviors. However, evidence for a common mechanism has been lacking. Stereotypies in human mental disorders are indicative of profound brain dysfunction involving the basal ganglia, and are associated with pervasive voluntary-motor impairments and psychological distress. Here we show that stereotypy in captive Orange-Wing Amazon Parrots (Amazona amazonica) is correlated with poor performance on the same psychiatric task (the 'gambling task') as stereotypy in autistic and schizophrenic patients. The task measures recurrent perseveration-the tendency to inappropriately repeat responses. Thus, the more stereotypy a parrot performed, the more likely it was to inappropriately repeat itself from trial-to-trial on the task; and the more rapidly it made repeated, but not switched, responses. These results parallel the executive motor impairments seen in human patients, and therefore suggest that, like in human patients, stereotypy in caged parrots reflects a general disinhibition of the behavioral control mechanisms of the dorsal basal ganglia. If this result holds true in other laboratory species, stereotypic animals are likely to be of questionable utility in behavior, neuroscience, and neuropharmacological experiments. In humans, stereotypies and obsessive-compulsive behaviors are considered to be mutually exclusive categories of behavior, with different neural substrates, and different treatment strategies. These results, therefore, suggest that the pharmacological treatment of stereotypies in veterinary medicine based on the assumption that they are equivalent to human Obsessive-Compulsive Disorder may be inappropriate. As stereotypies in captive animals develop in response to the captive environment, these results also emphasize the role that the environment may play in eliciting or exacerbating stereotypy in human patients. Finally, by parallel to human patients, there is a potential psychological distress in animals showing these behaviors. PMID- 14529812 TI - Non-elemental processing in olfactory discrimination tasks needs bilateral input in honeybees. AB - In patterning discriminations, animals have to differentiate a compound stimulus AB from each of its elements A and B. In positive patterning (PP), the compound is reinforced whilst the single elements are non-reinforced. In negative patterning (NP), single elements are reinforced whilst the compound is non reinforced. Using olfactory conditioning of the proboscis extension response (PER), we asked whether honeybees (Apis mellifera) can solve these patterning problems when odorants are given unilaterally as well as bilaterally to the antennae. Separating the olfactory input space of bees into two independent zones using plastic walls placed between the antennae, we conditioned bees in PP and NP procedures, with input on one side, on both sides, or in an ambiguous problem where bees had to solve PP on one side and NP on the other side. We found that bees with simultaneous bilateral input solve both patterning tasks efficiently. In contrast, PP but not NP was learned by bees receiving unilateral olfactory input. Bees subjected to the ambiguous NP/PP problem only solved PP. As PP can be solved through mere elemental processes, but NP is critically dependent on the use of non-elemental learning processes, our results suggest that bilateral olfactory input is necessary for non-elemental processing to take place in the bee brain. PMID- 14529814 TI - The effect of cumulative experience on the use of elemental and configural visual discrimination strategies in honeybees. AB - We addressed the question of whether the amount of individual experience determines the use of elemental or configural visual discrimination strategies in free-flying honeybees Apis mellifera. We trained bees to fly into a Y-maze to collect sucrose solution on a rewarded stimulus presented in one of the arms of the maze. Stimuli were colour disks, violet (V), green (G) or yellow (Y), which were of equal psychophysical salience for honeybees. Training followed an A+, BC+ design, followed by an AC versus BC test. Training consisted of 6 (3 A+ and 3 BC+), 20 (10 A+ and 10 BC+) or 40 (20 A+ and 20 BC+) acquisition trials. Elemental models of compound processing predict a preference for the non-trained stimulus AC while configural models predict a preference for the trained stimulus BC. Our results show that increasing the number of acquisition trials results in a change of the internal representation of stimuli. After six training trials, bees favoured an elemental strategy and preferred AC to BC during the tests. Generally, increasing the number of training trials resulted in an increase of the choice of BC. Thus, short training favoured processing of the compound as the sum of its elements (elemental account) while long training favoured its processing as being different from the sum of its elements (configural account). Additionally, we observed that the change in stimulus processing was also influenced by stimulus similarity. Colour perceptual similarity favoured configural processing with increasing experience. PMID- 14529813 TI - Anxiety-related behavior and densities of glutamate, GABAA, acetylcholine and serotonin receptors in the amygdala of seven inbred mouse strains. AB - The amygdala is a brain region involved in the regulation of anxiety-related behavior. The purpose of this study was to correlate anxiety-related behavior of inbred mouse strains (BA//c, BALB/cJ, C3H/HeJ, C57BL/6J, CPB-K, DBA/2J, NMRI) to receptor binding in the amygdala. Binding site densities of receptors (NMDA, AMPA, kainate, GABA(A), serotonin, muscarinergic M(1)-M(2)) were measured with quantitative receptor autoradiography using tritiated ligands. Measurements of fear-sensitized acoustic startle response (ASR; induced by footshocks), elevated plus maze (EPM) behavior and receptor binding studies showed differences between the strains except for AMPA and muscarinergic M(2) receptors. Factor analysis revealed a Startle Factor with positive loadings of the density of serotonin and kainate receptors, and the amplitudes of the baseline and fear-sensitized ASRs. A second Anxiety-related Factor only correlated with the fear-sensitized ASR and anxiety parameters on the EPM but not receptor densities. There were also two General Activity Factors defined by (negative) correlations with entries to closed arms of the EPM. Because the density of NMDA and muscarinergic M(1) receptors also correlated negatively with the two factors, these receptors had a positive effect on general activity. In contrast, correlations of GABA(A), serotonin, and kainate receptors had the opposite sign as compared to closed arm entries. It is concluded that hereditary variations in the amygdala, particularly in kainate and serotonin receptors, play a role for the baseline and fear sensitized ASR, whereas the general activity is influenced by many neurotransmitter receptor systems. PMID- 14529815 TI - Long-term effects of BIBN-99, a selective muscarinic M2 receptor antagonist, on improving spatial memory performance in aged cognitively impaired rats. AB - Aged Long-Evans rats were screened for spatial memory deficits using the Morris water maze task. Rats found to have impaired performance on the task (aged impaired, AI) were then treated with a selective muscarinic M2 receptor antagonist, 5,11-dihydro-8-chloro-11-[[4-[3-[(2,2-dimethyl-1 oxopentyl)ethylamino]propyl]-1-piperidinyl]acetyl]-6H-pyrido[2,3 b][1,4]benzodiazepin-6-one (BIBN-99; 0.5 mg/kg, s.c.), for 3 successive days while receiving additional water maze training. BIBN-99 significantly improved performance in the task during the 3 days of drug treatment. Treatment was then ceased for the remainder of the study and rats were tested again in the water maze on days 10, 17, and 24. Compared to vehicle-treated rats, enhanced performance was observed in the AI rats that had previously been treated with BIBN-99. These results indicate that BIBN-99 enhances spatial learning in AI animals and that enhanced (or long-term) memory persists in the absence of the drug. In a second experiment, a 2-month delay was imposed in between the original water maze screening and the drug treatment regime. Again, BIBN-99 significantly improved performance in AI rats. This latter study suggests that reference memory does not decay, even in an AI animal that had displayed poor learning following original water maze screening. Together, these studies help provide further insight into possible mechanism(s) of reference memory and its potential clinical usefulness. PMID- 14529816 TI - Emotional instability but intact spatial cognition in adenosine receptor 1 knock out mice. AB - Several lines of evidence point to the involvement of adenosine in the regulation of important central mechanisms such as cognition, arousal, aggression and anxiety. In order to elucidate the involvement of the adenosine A1 receptor (A1AR) in spatial learning and the control of exploratory behaviour, we assessed A1AR knockout mice (A1AR-/-) and their wild-type littermates (A1AR+/+) in a place navigation task in the water maze and in a battery of forced and free exploration tests. In the water maze, A1AR-/- mice showed normal escape latencies and were indistinguishable from controls with respect to measures of spatial performance during both training and probe trial. But despite normal performance they showed increased wall hugging, most prominently after the relocation of the goal platform for reversal training. Quantitative analysis of strategy choices indicated that wall hugging was increased mainly at the expense of chaining and passive floating, whereas the frequency of trials characterised as direct swims or focal searching was normal in A1AR-/- mice. These results indicate intact spatial cognition, but mildly altered emotional reactions to the water maze environment. In line with this interpretation, A1AR-/- mice showed normal levels and patterns of activity, but a mild increase of some measures of anxiety in our battery of forced and free exploration paradigms. These results are in line with findings published using a genetically similar line, but demonstrate that the magnitude of the changes and the range of affected behavioural measures may vary considerably depending on the environmental conditions during testing. PMID- 14529817 TI - Behavioral alterations in mice deficient for the extracellular matrix glycoprotein tenascin-R. AB - We investigated the behavior of mice deficient for the extracellular matrix (ECM) glycoprotein tenascin-R (TN-R) in comparison to their wild-type (WT) littermates. A longitudinal study including tests for exploration and anxiety, motor coordination and cognition was carried out. Mice were tested at different ages, ranging from 3 weeks to 11 months and under different housing conditions. TN-R deficient mice displayed decreased motivation to explore and an increased anxiety profile in the free choice open field (FCOF), open field (OF) and elevated plus maze (EPM) tests. Moreover, the anxiety level of TN-R deficient mice was more strongly influenced by environmental factors as compared to WT littermates. TN-R deficient mice showed motor coordination impairments in the wire hanging, Rotarod and pole test. Thus TN-R ablation leads to an altered behavioral phenotype in mice that may negatively affect their fitness under natural conditions. PMID- 14529818 TI - Brain injury and neurofunctional deficit in neonatal mice with hypoxic-ischemic encephalopathy. AB - Birth asphyxia accounts for the majority of developmental motor and cognitive deficits. Studies were undertaken to develop a reproducible murine model of perinatal hypoxic-ischemic encephalopathy (HIE) which would permit both anatomic and neurofunctional quantification of injury. Short-term neurofunctional outcomes consisted of three developmental reflexes (righting, cliff aversion and geotaxis) assessed in 7-day-old mouse pups 24 h after unilateral carotid artery ligation followed by inhalation of 8% oxygen. Cerebral infarct volume was dependent on duration of hypoxia, being approximately 2.5-fold greater with longer (60 min) versus shorter (30 min) hypoxia exposure (P=0.001). All three sensorimotor neonatal reflexes assessed at 24 h after HIE injury correlated significantly with long-term neurofunction evaluated using a water-maze test of navigational learning and memory assessed 8 weeks later in the same animals. Cerebral atrophy, a delayed consequence of HIE in this model, also correlated strongly with water maze performance (r=0.86, P=0.002). These data demonstrate for the first time that murine neonatal sensorimotor reflex performance can be rigorously quantified in the acute phase of perinatal HIE and has strong predictive value not only for anatomic extent of cerebral injury, but also for long-term neurofunctional outcome. PMID- 14529819 TI - Long-Evans and Sprague-Dawley rats have similar skilled reaching success and limb representations in motor cortex but different movements: some cautionary insights into the selection of rat strains for neurobiological motor research. AB - Many rat strains are used for neurobiological studies of nervous system function and behavior. The most widely used strain for studies of the neural basis of movement is the out bred, pigmented Long-Evans strain, while the most widely used strains for the study of movement impairments in neurological disease are out bred albino rats, including Sprague-Dawley rats. Although previous research has indicated that there are strain differences in skilled movements displayed by different rat strains, there has been no explicit comparison of the Long-Evans and Sprague-Dawley strains. This was the purpose of the present study. The rats were video recorded as they learned to reach for single food pellets and the video records were subjected to frame-by-frame analysis. Component movements of reaching were scored using a system derived from Eshkol-Wachman Movement Notation (EWMN). The quality of movements was described using Laban Movement Analysis (LMA). Forelimb representations in motor cortex were defined electrophysiologically. Acquisition scores and success in reaching did not differ between the two strains, nor did the topographical representation of the forelimb in motor cortex. Long-Evans and Sprague-Dawley rats did differ in the movements used for reaching and on the quality of their movements. The movements of Sprague Dawley rats resembled the movements of Long-Evans rats with motor system injury. That rat strains can show both quantitative and qualitative differences in movement is useful for the understanding of the genetic, neural, and behavioral organization of the motor system. The results are also relevant to the question of appropriateness of particular rat strains for studies of neurological diseases and the effects of albinism on motor behavior, and suggest that some of the most widely used rat strains for neurological investigations may be less than appropriate. PMID- 14529820 TI - The duration of response inhibition in the stop-signal paradigm varies with response force. AB - In a previous study, we have found that the speed of stopping a response is delayed when response readiness is reduced by cuing the probability of no-go trials [Acta Psychol. 111 (2002) 155]. Other investigators observed that responses are more forceful when the probability to respond is low than when it is high (e.g. [Quart. J. Exp. Psychol. A 50 (1997) 405]). In this study, the hypothesis was tested that low probability responses are more forceful than high probability responses and that these responses are more difficult to stop. Subjects performed on a choice reaction task and on three tasks with respectively 100%, 80%, and 50% response probabilities. Stop signals were presented on 30% of the trials, instructing subjects to withhold their response. Response force on non-signal (go) trials and the duration of response inhibition on signal (stop) trials increased as response probability decreased. This pattern of findings was interpreted to support the hypothesis predicting that stopping is more difficult when response readiness is low than when it is high. PMID- 14529821 TI - Large systematic deviations in a bimanual parallelity task: further analysis of contributing factors. AB - Previous studies showed that what subjects haptically perceive as parallel deviates largely from what is actually physically parallel [Perception 28 (1999) 1001; Acta Psychol. 109 (2002) 25; Perception 28 (1999) 781]. It also turned out that the deviations were strongly subject-dependent. It was hypothesized that what is haptically parallel is decided in a frame of reference intermediate to an allocentric and an egocentric one. The purposes of the present study were to collect more evidence for this hypothesis and to investigate the factor(s) that determines the specific weighting between the two reference frames. We found a highly significant reversal of a haptic oblique effect (in context: larger systematic deviations for oblique orientations) for subjects with large deviations. This reversal provides convincing evidence that an intermediate frame of reference is used for the decision of haptic parallelity. Contrary to common expectation, several factors that might have been of influence on the weighting of the two frames of reference, such as arm length, arm span, shoulder width, turned out to be irrelevant. Surprisingly, the only factors that seem to be of influence are gender and job experience or education. PMID- 14529822 TI - An empirical investigation of the visual rightness theory of picture perception. AB - This research subjected the visual rightness theory of picture perception to experimental scrutiny. It investigated the ability of adults untrained in the visual arts to discriminate between reproductions of original abstract and representational paintings by renowned artists from two experimentally manipulated less well-organized versions of each art stimulus. Perturbed stimuli contained either minor or major disruptions in the originals' principal structural networks. It was found that participants were significantly more successful in discriminating between originals and their highly altered, but not slightly altered, perturbation than expected by chance. Accuracy of detection was found to be a function of style of painting and a viewer's way of thinking about a work as determined from their verbal reactions to it. Specifically, hit rates for originals were highest for abstract works when participants focused on their compositional style and form and highest for representational works when their content and realism were the focus of attention. Findings support the view that visually right (i.e., "good") compositions have efficient structural organizations that are visually salient to viewers who lack formal training in the visual arts. PMID- 14529823 TI - Perceptual dimensions of tactile textures. AB - The present study investigated the perceptual dimensions of everyday tactile textures and the semantics associated with touch experiences. In Experiment 1, the nature of the tactile descriptors present in the memory of 40 volunteers was investigated. Results suggested the existence of a limited but consensual tactile repertory. In Experiment 2, 20 volunteers used a free-sorting task to sort 24 car seat ' materials, with different tactile properties, on the basis of perceived similarity. Sorted groups were then characterized further by producing verbal labels. Multidimensional scaling and correspondence analysis indicated that the tactile texture space did not exceed four dimensions. The hypothesized perceptual dimensions were soft/harsh, thin/thick, relief and hardness. Further evidence for the relevance of these dimensions was provided in Experiment 3, in which participants were asked to evaluate the stimuli according to unipolar adjective scales corresponding to the previously derived dimensions. PMID- 14529824 TI - Repetition priming effects from attended vs. ignored single words in a semantic categorization task. AB - The present research examines priming effects from a centrally presented single prime word to which participants were instructed to either attend or ignore. The prime word was followed by a single central target word to which participants made a semantic categorization (animate vs. inanimate) task. The main variables manipulated across experiments were attentional instructions (attend vs. ignore the prime word), presentation duration of the prime word (20, 50, 80 or 100 ms), prime-target stimulus onset asynchrony (SOA; 300 vs. 800 ms), and temporal presentation of instructions (before vs. after the prime word). The results showed (a) a consistent interaction between attentional instructions and repetition priming and (b) a qualitatively different ignored priming pattern as a function of prime duration: reduced positive priming (relative to the attend instruction) for prime exposures of 80 and 100 ms, and reliable negative priming for the shorter prime exposures of 20 and 50 ms. In addition (c), the differential priming pattern for attend and ignore trials was observed at a prime target SOA of 800 ms (but not at a shorter 300-ms SOA) and only when instructions were presented before the prime word. Methodological and theoretical implications of the present findings for the extant negative priming literature are discussed. PMID- 14529825 TI - Treatment of rat gliosarcoma brain tumors by HSV-based multigene therapy combined with radiosurgery. AB - Our laboratory has employed replication-defective herpes simplex virus type 1 gene transfer vectors for treatment of animal models of human malignant glioblastoma. The base vectors were defective for the immediate early (IE) genes ICP4, ICP27, and ICP22 but expressed the IE gene ICP0, which can arrest tumor cell division, and an IE thymidine kinase (alpha-tk) gene construct that mediates suicide gene therapy (SGT) in the presence of ganciclovir (GCV). Previously, we reported that SGT using ICP0/alpha-tk vectors in nude mouse models of glioblastoma was improved by coexpression of the gap-junction-forming protein connexin43 (Cx43) or human tumor necrosis factor alpha (TNF alpha). We also showed that further gains in therapeutic outcome could be achieved by combining TNF alpha-enhanced SGT with gamma-knife radiosurgery (GKR). To expand these observations, we have first repeated these studies in immunocompetent rats with brain tumors derived from implanted 9L gliosarcoma cells and second compared the most efficient vector from this study with a new recombinant vector, NUREL-C2, which expressed both TNF alpha and Cx43 along with ICP0 and alpha-tk. Results from the first part indicated that our ICP0/alpha-tk/TNF alpha vector in combination with GKR provides an effective therapy although this treatment was not statistically better than GKR combined with the ICP0/alpha-tk/Cx43 vector. Our observations in the second part suggested that NUREL-C2 may be more effective than the ICP0/alpha-tk/TNF alpha vector in combination treatments with GCV (P = 0.08) or GCV plus GKR (P = 0.10). GKR significantly enhanced the efficacy of NUREL-C2/GCV treatment (P = 0.02) as well as other virus/GCV treatments (P < or = 0.05). Conversely, the efficacy of GKR was significantly improved by both the ICP0/alpha-tk/TNF alpha vector and NUREL-C2 in combination with GCV (P = 0.02 and P < 0.01, respectively). Together these results indicate that NUREL-C2 may be an attractive candidate for Phase I gene-therapy safety studies in patients with recurrent malignant glioblastoma. PMID- 14529826 TI - HSV-1 therapy of primary tumors reduces the number of metastases in an immune competent model of metastatic breast cancer. AB - The HSV-1 1716 mutant virus and similar oncolytic herpesviruses deficient in the gamma 34.5 neurovirulence gene are able to reduce the growth of tumors in mice. Here we demonstrate that HSV-1 1716 therapy moderately reduced the growth of tumors of the highly malignant, spontaneously metastasizing 4T1 mouse mammary carcinoma model. This moderate effect on 4T1 tumor growth was likely due to poor replication kinetics of HSV-1 1716 in 4T1 cells. Interestingly, HSV-1 therapy of the primary tumor increased the survival time of mice. Coincident with this increase was a reduction in metastases as determined by quantification of the number of metastatic cells in the lungs. HSV-1 therapy of the primary tumor was also able to reduce the establishment of a second challenge of 4T1 tumors. Moreover, infiltrates of both CD4(+) and CD8(+) T cells were detected in HSV-1 1716-treated tumors. An important role for the T cell infiltrates was confirmed when HSV-1 therapy did not reduce the growth of 4T1 tumors in SCID mice. Collectively, these results demonstrate that an HSV-dependent anti-tumor immune response is required for the reduction in primary 4T1 tumor growth and for the reduction in the establishment of metastases in this tumor model. PMID- 14529827 TI - Interleukin (IL)-21 and IL-15 genetic transfer synergistically augments therapeutic antitumor immunity and promotes regression of metastatic lymphoma. AB - IL-21 supports proliferation of mature T and B cells and facilitates expansion and maturation of natural killer (NK) cells in synergy with IL-15. However, the biological implications of IL-21 in vivo have not been fully elucidated. IL-21 and IL-15 expression plasmids were intravenously injected under high pressure into the tail veins of mice, which were subsequently challenged by an intravenous injection of RLmale1 lymphoma cells. The IL15 gene transfection significantly reduced the numbers of metastatic tumor foci in the liver. In contrast, when IL21 and IL15 genes were cotransfected, complete regression was achieved in 80% of the mice. The cytokine gene therapy was also performed in mice that had been intravenously inoculated with the tumor cells. Forty percent of mice that received a single injection of a mixture of cytokine genes successfully rejected the preestablished metastatic lymphoma and showed tumor-free survival for more than 300 days. IL-21 significantly elevated the cytotoxic T lymphocyte activity in the spleens of tumor-inoculated mice, while the two cytokines augmented NK killing activity in a synergistic manner. These results strongly suggest that the codelivery of IL-21 and IL-15 elicits powerful antitumor immune responses, resulting in marked therapeutic efficacy against metastatic tumors. PMID- 14529828 TI - Boosting with recombinant vaccinia increases HPV-16 E7-Specific T cell precursor frequencies and antitumor effects of HPV-16 E7-expressing Sindbis virus replicon particles. AB - Immunotherapy using the heterologous prime-boost regimen has emerged as an attractive approach for generating antigen-specific T-cell-mediated immune responses against tumors and infectious diseases. We have previously linked the Mycobacterium tuberculosis heat-shock protein 70 (HSP70) to the HPV-16 E7 antigen creating a chimera, E7/HSP70. We found that nucleic acid vaccines encoding E7/HSP70 can generate strong antitumor immunity. Recently, replication-defective Sindbis virus replicon particle vaccines have been considered as an important vector system for vaccine development. In this study, we assessed whether the combination of E7/HSP70 Sindbis virus replicon particles (SINrep5-E7/HSP70) and E7/HSP70 vaccinia (Vac-E7/HSP70) can further enhance E7-specific immune responses using sequential vaccination. We found that priming with SINrep5-E7/HSP70 and boosting with Vac-E7/HSP70 generated the highest number of E7-specific CD8(+) T cells and best antitumor effect compared to other combinations. Moreover, our data showed that at the dosage and route of immunization used in this study, mice treated with the Sindbis virus replicon particle prime-vaccinia boost regimen generated stronger antitumor responses compared to mice treated with the DNA prime-vaccinia boost vaccine regimen. Our results encourage the use of the Sindbis virus replicon particle prime-vaccinia boost regimen in future clinical trials. PMID- 14529829 TI - Transcriptional targeting of dendritic cells in gene gun-mediated DNA immunization favors the induction of type 1 immune responses. AB - Cutaneous dendritic cells (DC) are pivotal for the elicitation of antigen specific immune responses following gene gun-mediated biolistic transfection of the skin. We transcriptionally targeted transgene expression to DC using vectors containing the murine fascin promoter (pFascin) to control antigen production and compared the immune response elicited with conventional DNA immunization using plasmid constructs with the ubiquitously active CMV promoter (pCMV). Biolistic transfection with pFascin initiated a marked type 1 immune response characterized by the occurrence of a large population of IFN-gamma-producing T helper (Th) cells in spleen and draining lymph nodes. Consistently, immunoglobulin production was dominated by IgG2a antibodies. In contrast, the humoral response after repeated administration of pCMV was strongly enhanced and characterized by a type 2-like isotype pattern (IgG1 > IgG2a). Cytokine production analysis in vitro indicated compartmentalization of the immune response, revealing large numbers of IL-4-producing Th cells in the lymph nodes and dominant presence of IFN-gamma producing Th cells in the spleen. Biolistic transfection with pFascin, like immunization with pCMV, led to potent induction of cytotoxic T cells as was assessed by JAM test. Thus gene gun immunization with plasmids that focus transgene expression and antigen production specifically to DC propagates type 1 biased cellular immune responses. PMID- 14529830 TI - Macrophage apolipoprotein A-I expression protects against atherosclerosis in ApoE deficient mice and up-regulates ABC transporters. AB - The antiatherogenic effect of high-density lipoprotein (HDL) and its major protein component apolipoprotein A-I (apoA-I) has been largely attributed to their key roles in reverse cholesterol transport (RCT) and cellular cholesterol efflux. Substantial evidence shows that overexpression of human apoA-I reduces atherosclerosis in animal models. However, it is uncertain whether this protection is due to an increase in plasma HDL level or to a local effect in the artery wall. To test the hypothesis that expression of human apoA-I in macrophages can promote RCT in the artery wall, we used a retroviral construct expressing human apoA-I cDNA (MFG-HAI) to transduce ApoE(-/-) bone marrow cells and then transplanted these cells into ApoE(-/-) mice with preexisting atherosclerosis. ApoE(-/-) mice reconstituted with MFG-HAI marrow had a significant reduction (30%) in atherosclerotic lesions in the proximal aorta compared to control mice that received marrow expressing MFG parental virus. Peritoneal macrophages isolated from MFG-HAI mice showed a four- to fivefold increase in mRNA expression levels of both ATP-binding cassette (ABC) A1 and ABCG1 compared to controls. Our data demonstrate that gene transfer-mediated expression of human apoA-I in macrophages can compensate in part for apoE deficiency and delay the progression of atherosclerotic lesions by stimulating ABC-dependent cholesterol efflux and RCT. PMID- 14529831 TI - Adenoviral-delivered angiopoietin-1 reduces the infarction and attenuates the progression of cardiac dysfunction in the rat model of acute myocardial infarction. AB - In acute myocardial infarction (AMI), prognosis and mortality rate are closely related to the infarct size and the progression of postinfarction cardiac failure. Angiogenic gene therapy has presented a new approach for the treatment of AMI. Angiopoietin-1 (Ang1) is a critical angiogenic factor for vascular maturation and enhances vascular endothelial growth factor (VEGF)-induced angiogenesis in a complementary manner. We hypothesized that gene therapy using Ang1 for AMI might promote angiogenesis cooperatively with intrinsic VEGF, since high concentrations of circulating VEGF have been reported in AMI. To evaluate our hypothesis, we employed a rat AMI model and adenoviral Ang1 (HGMW-approved gene symbol ANGPT1) gene transfer to the heart. A significant increase in capillary density and reduction in infarct sizes were noted in the infarcted hearts with adenoviral Ang1 gene treatment compared with control infarcted hearts treated with saline or adenoviral vector containing the beta-galactosidase gene. Furthermore, the Ang1 group showed significantly higher cardiac performance in echocardiography (55.0% of ejection fraction, P < 0.05 vs control) than the saline or adenoviral controls (36.0 or 40.5%, respectively) 4 weeks after myocardial infarction. The adenoviral delivery of Ang1 during the acute phase of myocardial infarction would be feasible to attenuate the progression of cardiac dysfunction in the rat model. PMID- 14529832 TI - Long-term correction of bilirubin UDPglucuronyltransferase deficiency in rats by in utero lentiviral gene transfer. AB - Bilirubin is glucuronidated by bilirubin UDP-glucuronyltransferase (UGT1A1) before biliary excretion. Because bilirubin is toxic, patients with Crigler Najjar type I (CN), who have no UGT1A1 activity, suffer severe brain damage early in childhood. The Gunn rat is the model for CN type 1. Gunn rat fetuses were injected with 10(7) transducing units of UGT1A1 lentiviral vector at the end of the third trimester on embryonic day 19. Serum bilirubin of injected Gunn rats was lowered by 45% compared to untreated controls. This decrease was highly significant (P < 10(6)) and was sustained for more than a year. In treated Gunn rats, bilirubin glucuronides were present in bile and UGT1A1 protein was detected in tissue. Liver, intestine, stomach, pancreas, and other organs were transduced and mostly contained 1% or less vector copies per genome. Tissue distribution was variable among experimental animals but high transduction levels were seen in pancreas and intestine in most animals. Immunohistochemistry of these organs revealed transduction of pancreatic acinar cells and intestinal epithelium. Injection of a lentiviral UGT1A1 vector into third-trimester Gunn rat fetuses corrects the metabolic deficiency and mediates a reduction of serum bilirubin levels that would be therapeutic in humans. PMID- 14529833 TI - Phenotype correction of Fanconi anemia group A hematopoietic stem cells using lentiviral vector. AB - Fanconi anemia (FA) is an autosomal recessive disease characterized by progressive bone marrow failure due to defective stem cell function. FA patients' cells are hypersensitive to DNA cross-linking agents such as mitomycin C (MMC), exposure to which results in cytogenetic aberrations and cell death. To date Moloney murine leukemia virus vectors have been used in clinical gene therapy. Recently, third-generation lentiviral vectors based on the HIV-1 genome have been developed for efficient gene transfer to hematopoietic stem cells. We generated a self-inactivating lentiviral vector expressing the FA group A cDNA driven by the murine stem cell virus U3 LTR promoter and used the vector to transduce side population (SP) cells isolated from bone marrow of Fanconi anemia group A (Fanca) knockout mice. One thousand transduced SP cells reconstituted the bone marrow of sublethally irradiated Fanca recipient mice. Phenotype correction was demonstrated by stable hematopoiesis following MMC challenge. Using real-time PCR, one proviral vector DNA copy per cell was detected in all lineage-committed cells in the peripheral blood of both primary and secondary recipients. Our results suggest that the lentiviral vector transduces stem cells capable of self renewal and long-term hematopoiesis in vivo and is potentially useful for clinical gene therapy of FA hematopoietic cells. PMID- 14529834 TI - Direct comparison of RD114-pseudotyped versus amphotropic-pseudotyped retroviral vectors for transduction of rhesus macaque long-term repopulating cells. AB - Recently, RD114 (feline endogenous retrovirus envelope protein)-pseudotyped retroviral particles have been shown to transduce human NOD/SCID repopulating cells efficiently. In this study, we compared directly transduction of repopulating cells with RD114-pseudotyped vector to that with standard amphotropic vector in the rhesus macaque model. G-CSF/SCF-mobilized CD34(+) rhesus peripheral blood cells were cultured in the presence of SCF, Flt-3 ligand, and MGDF on Retronectin-coated flasks. To assess directly the ability of the two pseudotypes to transduce primitive cells, both vectors were added simultaneously to the target cells every 24 h, for a total of four exposures in 96 h. The cells were reinfused after the animals received 1000 cGy total body irradiation. At the end of transduction, gene marking efficiency of CFU was higher with amphotropic LNL6 vector (mean 88.4%) vs RD114-G1Na vector (mean 18.5%). After long-term engraftment in three animals, total neo gene marking levels were 4-5% in PBMNCs and 1.5-4% in granulocytes. The RD114-G1Na marking levels were consistently higher in granulocytes than in mononuclear cells, while amphotropic LNL6 marking levels were higher in PBMNCs than in granulocytes. The differential gene marking patterns suggest that RD114 and amphotropic vectors may target distinct progenitor or stem cell populations. There was no clear advantage for RD114 pseudotyped vectors in this predictive preclinical model in terms of overall long term marking levels; however, optimization of transduction conditions by increasing m.o.i. or inducing the receptor could potentially improve results with this novel vector system. PMID- 14529835 TI - Enforced adenoviral vector-mediated expression of HOXB4 in human umbilical cord blood CD34+ cells promotes myeloid differentiation but not proliferation. AB - Retroviral overexpression of the transcription factor HOXB4 results in a rapid increase in proliferation of murine hematopoietic stem cells both in vivo and in vitro. Therefore, we asked whether transient overexpression of HOXB4 would increase proliferation of human primitive hematopoietic progenitors. Transient overexpression of HOXB4 was generated in umbilical cord blood (CB) CD34(+) cells by a recombinant adenovirus (AdHOXB4) expressing HOXB4 together with the enhanced green fluorescent protein (GFP). Transduced, GFP(+) cells were cultured in serum free medium containing cytokines that primarily support the growth of primitive hematopoietic progenitors. In contrast to previous findings using retroviral overexpression of HOXB4, we did not observe any increase in proliferation of primitive progenitors or increased colony formation of clonogenic progenitors, including progenitor progeny from long-term culture-initiating cells following adenoviral vector overexpression of HOXB4 in CB CD34(+) cells. However, enforced expression of HOXB4 by the adenoviral vector significantly increased myeloid differentiation of primitive hematopoietic progenitors. Since retroviral vectors generate low and continuous levels of transgene expression in contrast to the high, transient levels generated by the adenoviral vector, our findings suggest that the high levels of HOXB4 expression generated by AdHOXB4 in human CB CD34(+) cells direct the cells toward a myeloid differentiation program rather than increased proliferation. PMID- 14529836 TI - Optimization of gene expression in nonactivated circulating lymphocytes. AB - Circulating lymphocytes are important target cells for the treatment of HIV related and autoimmune diseases and for stimulating anti-tumor immunity. To date, gene transfection of these nonactivated cells after intravenous delivery of viral or nonviral vectors remains low although these circulating cells are highly accessible. Optimized lentiviral vectors currently can transduce less than 10% of nonactivated circulating lymphocytes. Here we report transfection of up to 15% of these nonactivated cells using liposomes directed to human CCR5 displayed on the surface of helper T cells and macrophages in transgenic mice. Attachment of modified MIP-1 beta to the surface of DNA-liposome complexes increased gene delivery and expression in nonactivated circulating lymphocytes approximately sixfold. In vitro data using these complexes to transfect PM1 cells that have elevated levels of CCR5 supported our data obtained in vivo. Therefore, ligands that bind to cell surface receptors on circulating lymphocytes can be used with optimized systemic liposomes to increase transfection and gene expression in these cells without activation. PMID- 14529837 TI - Targeting expression of a transgene to the airway surface epithelium using a ciliated cell-specific promoter. AB - Many of the vectors being investigated for gene therapy utilize viral promoters or promoters from ubiquitously expressed genes (e.g., CMV, beta-actin). These promoters are active in many cell types and generally result in high levels of transgene expression. However, the use of these promoters for gene therapy of cystic fibrosis (CF) may produce undesirable effects by directing high levels of CFTR expression in cells that normally do not synthesize this protein. In contrast, a vector containing a ciliated cell-specific promoter and delivered to the lung would be active only in the ciliated cells that line the surface of the airways. Ciliated cells express CFTR and are in direct contact with the airway surface liquid normally regulated by CFTR. To develop a ciliated cell-specific promoter for CF gene therapy, we have characterized the promoter region of the FOXJ1 gene, a transcription factor required for ciliated cell differentiation. A fragment of the human FOXJ1 promoter region was inserted into an EGFP expression cassette and used to produce transgenic mice. Transgene-positive animals demonstrated strong EGFP expression in the ciliated cells of tracheal, bronchial, and nasal epithelium. Our results demonstrate that elements within the FOXJ1 promoter region are sufficient to target expression of transgenes to ciliated cells and may be useful for gene therapy of CF. PMID- 14529838 TI - Transfection efficiency and toxicity following delivery of naked plasmid DNA and cationic lipid-DNA complexes to ovine lung segments. AB - We defined, using a novel large animal model system, the acute pathologic response to localized pulmonary administration of either naked plasmid DNA (pDNA) or cationic lipid-pDNA complexes (pDNA:GL67) and related such responses to concomitant indicators of transfection efficiency, namely levels of chloramphenicol acetyl transferase (CAT) protein and mRNA in specific lung tissue compartments. We instilled doses of 0.2, 1, and 5 mg pDNA to spatially distinct lung segments in six anesthetized sheep and doses of 0.2, 1, and 5 mg pDNA:GL67 to a further six sheep. Twenty-four hours after gene delivery the sheep were euthanized and necropsy examination with sampling of relevant tissues was carried out. Levels of plasmid-derived CAT-specific mRNA and CAT protein in samples derived from segments treated with either pDNA or pDNA:GL67 increased in relation to the administered dose. Levels of mRNA and protein expression were greater for pDNA:GL67 than for pDNA alone. A significant correlation was observed between mRNA and protein expression in samples derived from airways treated with pDNA:GL67. Histopathological changes following administration of both pDNA and pDNA:GL67 were characterized by a neutrophilic inflammation predominantly oriented on airways. The severity of the inflammatory response appeared to correlate with the administered dose of DNA and was generally more severe for pDNA:GL67. PMID- 14529839 TI - Helper-Independent Sleeping Beauty transposon-transposase vectors for efficient nonviral gene delivery and persistent gene expression in vivo. AB - Transposon-based vectors represent promising new tools for chromosomal transgene insertion and establishment of persistent gene expression in vivo. Here, we report the development of helper-independent transposon-transposase (HITT) vectors, which contain on single plasmids (i) a Sleeping Beauty (SB) transposon containing the transgene and (ii) a SB transposase expression cassette. To obtain an optimal level of transposase expression from HITT vectors, we determined the relative strength of a panel of different promoters in mouse liver and used these promoters to drive transposase expression from injected HITT vectors carrying a human alpha(1)-antitrypsin (hAAT) expression cassette flanked by transposon inverted repeats. By correlating promoter strength with stabilized serum hAAT levels, a narrow expression window supporting high-level transposition in the liver was defined. Peak levels of long-term gene expression were obtained with promoters 30- to 40-fold less active than CMV in mouse liver, whereas reduced stable levels of hAAT were detected with both weaker and stronger promoters. Injected HITT vectors induced transposase-dependent insertion of transposon DNA into the genome of at least 5-6% of transfected hepatocytes, generating levels of persistent hAAT expression that were 2- to 4-fold higher than with an optimized two-plasmid approach. In addition, we show that HITT vectors carrying a human factor IX (hFIX)-containing transposon support (i) long-term hFIX expression in normal mice and (ii) partial phenotypic correction in a mouse model of hemophilia B. SB-based HITT vectors represent a major advance in the establishment of persistent transgene expression from nonviral gene delivery systems and should prove useful for gene transfer to tissues or cell types in which transfection efficiencies are low. PMID- 14529840 TI - Generation of transgenic mice using lentiviral vectors: a novel preclinical assessment of lentiviral vectors for gene therapy. AB - Lentiviral vectors have become attractive delivery vehicles for gene therapy investigators. Specifically, the ability of lentiviral vectors to integrate into nondividing cells and provide stable and long-term gene expression in vivo is a desirable attribute of gene therapy approaches. We report here a simple method for generating transgenic mice using lentiviral vectors, which could be useful models for gene therapy. After removal of the zona pellucida, fertilized eggs were co-incubated with oncoretroviral or lentiviral vectors. The resulting blastocysts were transferred into uteri of pseudo-pregnant females. In both cases, around 60-70% of founder pups were transgenic as determined by PCR analysis. Southern blot analysis revealed that the transgenes were integrated at different genetic loci and transmitted through the germ line. Most of the transgenes delivered by lentiviral vectors were expressed in transgenic mice, although those delivered by oncoretroviral vectors were completely silenced. When the upstream sequences of the rhodopsin gene and the red pigment gene were used as tissue-specific promoters, consistent enhanced green fluorescent protein (EGFP) expression was observed in rod and cone photoreceptor cells, respectively, in retina. However, mice generated with the corneal epithelium-specific keratin 12 promoter displayed EGFP expression not only in cornea but also in other tissues of the mouse. We conclude that the generation of transgenic mice using lentiviral vectors is a simple and robust method to evaluate the promoter specificity in lentiviral vectors in vivo prior to undertaking a gene therapy strategy. PMID- 14529841 TI - Global analysis of cellular transcription following infection with an HIV-based vector. AB - We have examined the changes in cellular transcription resulting from infection with HIV-based vectors. Previous work suggested that the incoming viral genome may under some circumstances be detected as DNA damage, so to explore this possibility, we compared the transcriptional response to infection with an HIV based vector to the response to treatment with the DNA-damaging agent etoposide. Expression levels of about 12,000 cellular RNA transcripts were determined in a human B-cell line at different times after either treatment. Statistical analysis revealed that the infection with the lentivirus vector resulted in quite modest changes in gene expression. Treatment with etoposide, in contrast, caused drastic changes in expression of genes known or inferred to be involved in apoptosis. Statistically significant though subtle parallels in the cellular transcriptional responses to etoposide treatment and HIV-vector infection could be detected. Several further data sets analyzing infections with HIV-based vectors or wild type HIV-1 showed similar modest effects on cellular transcription and very modest parallels among different data sets. These findings establish that HIV vector or HIV-1 infection has remarkably little effect on cellular transcription. The statistical methods described here may be of wide use in mining microarray data sets. Our observations support the idea that gene therapy with HIV-based vectors should not be particularly toxic to cells due to disruption of cellular transcription. PMID- 14529842 TI - Metabolically biotinylated adenovirus for cell targeting, ligand screening, and vector purification. AB - Development of cell-targeting vectors is an important focus for gene therapy. While some ligands can be genetically inserted into virus capsid proteins for cell targeting, for many ligands, this approach can disrupt either ligand function or vector function. To address this problem for adenovirus type 5 vectors, the fiber capsid protein was genetically fused to a biotin acceptor peptide (BAP). Adenovirus particles bearing this BAP were metabolically biotinylated during vector production by the endogenous biotin ligase in 293 cells to produce covalently biotinylated virions. The resulting biotinylated vector could be retargeted to new receptors by conjugation to biotinylated antibodies using tetrameric avidin (K(d) = 10(-15) M). The biotinylated vector could also be purified by biotin-reversible binding on monomeric avidin (K(d) = 10(-7) M). Finally, this vector was used as a ligand screening platform for dendritic cells in which a variety of structurally diverse protein, carbohydrate, and nucleic acid ligands were easily added to the vector using the biotin-avidin interaction. This work demonstrates the utility of metabolically biotinylated viruses for ligand screening, vector targeting, and virus purification applications. PMID- 14529844 TI - Immunosuppressive management following intestinal transplantation in adult patients. PMID- 14529845 TI - Role of genetic testing in liver transplantation for Wilson's disease. PMID- 14529846 TI - Daily interferon alpha-2B and ribavirin combination therapy for liver transplant patients with chronic hepatitis C infection. PMID- 14529847 TI - Differential diagnosis of acute rejection versus hepatitis reinfection. PMID- 14529848 TI - Differential diagnosis of cytomegalovirus infection and acute rejection by serum CC-chemokine measurement after orthotopic liver transplantation. PMID- 14529849 TI - Risks and benefits of nucleosides before and after liver transplantation. PMID- 14529850 TI - Living donor kidney transplantation: chance for the recipient--financial risk for the donor? AB - BACKGROUND: With living donation, in addition to the medical risk, the financial risk for the donor is essential, especially in case of complications that potentially can led to disability and loss of work. We report the experiences of those who have donated a kidney in our transplant center. METHODS: We contacted 80 donors who donated a kidney at least 6 months prior to evaluation: 72% answered 33 questions. [mean age: 54 +/- 10 (33-75) years; 69% living related, 31% unrelated]. RESULTS: Of the 80 donors contacted, 91% (53) reported to have no financial expenses due to donation; 9% (5) had expenses, but only few of them clarified exact amount. One donor had to borrow money to cover the lack when he was unable to perform his job. Another claimed the disparity between normal salary and payment from insurance company as a financial expense. Evaluation procedure prior to donation was organized variously: some donors were on holiday while evaluated, some officially were ill, others had to take off some days without payment. None of the donors lost his or her job due to donation. CONCLUSION: The financial risk of living donation is theoretically well covered by different insurances. However, some of the donors had to cover some expenses by themselves. Fortunately, so far in our center no major complications occurred and all donors went home in good health after donation. If costs are covered when a healthy donor loses his or her ability to work due to donation remains unclear since no donor has experienced this problem. PMID- 14529851 TI - Laparoscopic living donor nephrectomy: from alternative to standard procedure. PMID- 14529852 TI - Living donor nephrectomy in an open technique; a long-term analysis of donor outcome. PMID- 14529853 TI - Expanding the donor pool using marginal organs: single-center experience with 36 double-kidney transplantations in 5 years. PMID- 14529854 TI - ATG versus basiliximab induction therapy in renal allograft recipients receiving a dual immunosuppressive regimen: one-year results. PMID- 14529855 TI - Portal venous and enteric exocrine drainage in simultaneous pancreas-kidney transplantation: experience with 70 consecutive transplant recipients. PMID- 14529857 TI - Urological complications of renal transplantation: a series of 900 cases. PMID- 14529856 TI - Patients with steroid refractory acute vascular rejection develop agonistic antibodies targeting angiotensin II type 1 receptor. PMID- 14529858 TI - Criteria for the assessment of the donor heart. PMID- 14529859 TI - Noninvasive surveillance strategy for early identification of heart transplant recipients with possible coronary stenoses. PMID- 14529860 TI - Primary pulmonary hypertension: survival benefits of therapy with prostacyclin analogs and transplantation. PMID- 14529861 TI - Lack of impact of donor sodium levels on outcome after heart transplantation. PMID- 14529862 TI - Evidence-based immunosuppression after pediatric renal transplantation--a dream? PMID- 14529863 TI - C2 monitoring and absorption profiling of cyclosporine for optimization of immunosuppressive therapy in pediatric renal transplant recipients. PMID- 14529864 TI - Protocol biopsies as predictors of chronic allograft nephropathy. PMID- 14529865 TI - Rapamycin treatment at immunosuppressive doses affects tumor blood vessel circulation. PMID- 14529866 TI - Influence of a selective endothelin(A) receptor antagonist on the quantitative mRNA expression and the immunohistochemistry of vasoactive mediators after pancreas transplantation. PMID- 14529867 TI - Heat-exposed pig islets are protected from inflammatory mediators but express proapoptotic proteins. PMID- 14529868 TI - Insulin-positive cells in vitro generated from rat bone marrow stromal cells. PMID- 14529869 TI - Pretreatment of isolated islets with caspase-3 inhibitor DEVD increases graft survival after xenotransplantation. PMID- 14529870 TI - Pediatric donor organs for pancreas transplantation: an underutilized resource? PMID- 14529872 TI - Factors that impact on immediate graft function in patients after renal transplantation. AB - Kidney transplantation has become therapy of choice for patients with end-stage renal failure. However, many factors may cause graft rejection or delayed graft function, both of which decrease the prognosis for graft survival. For transplantologists the most important endeavor is to eliminate factors responsible for shortening graft function and to find those predictive of immediate graft function. The aim of the study was to investigate which factors influence early graft function. We retrospectively reviewed 442 renal transplant patients performed between 1990 and 1995 in two Szezecin units. All patients received an identical immunosuppressive drug schedule. Three hundred twelve patients who displayed immediate graft function were included in the study group to analyze donor and recipient age and sex, etiology of ESRD, HLA compatibility AB0 and Rh compatibility cold ischemia time, warm ischemia time, antileukocytes antibodies (PRA), and period of dialysis therapy before transplantation. We observed statistical significance for HLA and AB0 compatibility, younger donor age, and shorter cold ischemia time as the most important factors predictive of early graft function and an improved prognosis for graft survival. PMID- 14529873 TI - Cytokine gene expression in kidney allograft donor biopsies after cold ischemia and reperfusion using in situ RT-PCR analysis. AB - It was previously reported that ischemia-reperfusion injury initiates an inflammatory response and may significantly affect the transplanted organ function. The aim of this study was to assess changes of intragraft cytokine mRNA expression in kidneys after cold ischemia (CI) and following reperfusion. We examined mRNA of a product of activated T lymphocytes (IFN-gamma) and a monocyte product (IL-6). Eleven kidneys were transplanted after CI time ranging from 16 to 39 hours. Renal needle core biopsies were obtained from donors after cold ischemia and approximately after 20 minutes of reperfusion. Tubular and glomerular expression of IFN-gamma and IL-6 mRNA were assessed using semiquantitative evaluation of the RT-PCR in situ. After reperfusion an intense increase of IL-6 mRNA expression was observed in four specimens, a slight increase was noticed in five specimens, and a very slight decrease in two specimens. Changes in IL-6 mRNA expression were limited only to tubules. In contrast, the glomerular and tubular mRNA expression of IFN-gamma and glomerular of IL-6 remained stable. Mean CI time for patients with an intense increase was higher than for patients with a slight increase and with the decrease of IL-6 mRNA expression (32.0 +/- 6.8 vs 25.2 +/- 7.3 and 26.0 +/- 5.7 hours). Our results suggest that early inflammatory changes at the time of implantation of renal allografts depends mainly on monocyte/macrophage-associated products. The observed intensity of their expression in tubules was connected to longer CI time. PMID- 14529874 TI - Variations in temperature of the donor kidney during cold ischemia time and subsequent assessment of reperfusion using the application of thermovision camera. AB - Cold ischemia time (CIT) and graft reperfusion events are important nonimmunological factors that influence kidney graft function. The optimal temperature for the organs during CIT ranges from 4 degrees C to 8 degrees C. However, preservation of the designated temperature is usually not controlled during standard storage procedures. Aspects of initial graft reperfusion are usually assessed indirectly. Better evaluation of the effectiveness of the early blood supply may improve the surgical outcome. The aim of the study was to monitor the temperature during CIT in the kidney and surrounding area and subsequently to assess graft reperfusion using thermography. Temperature values of the area surrounding the kidney were registered during 8 organ procurements. We observed that the area surrounding the kidney displayed the optimal temperature range only during the first 5 minutes of CIT; later the temperature oscillated between 1 degrees C and 3.5 degrees C. The study proved that the temperature of the preservation fluid is frequently below prescribed. Analysis of the thermograms of 40 transplanted kidneys with the use of a thermovision camera ThermaCAM SC500 showed usefulness of this method to assess reperfusion and blood distribution in the transplanted kidney. PMID- 14529875 TI - Do all well-differentiated thyroid cancers constitute a definite contraindication to obtaining organs for transplantation? A case report. AB - In this case a thyroid gland tumor was diagnosed with fine needle aspiration (FNA) in a 34-year-old female donor of a liver fragment for living related liver transplantation. This diagnosis disqualified her as a donor. The increased incidence of thyroid cancer in Poland presents the possibility of their occurrence in potential donors. Well-differentiated thyroid papillary carcinomas larger than 1 cm in diameter, as well as follicular and medullary carcinomas (regardless their size and or clinical staging), present absolute contraindication to donation. Papillary microcarcinoma restricted to the thyroid gland (with no metastases in local lymph nodes) because of its specific behavior and almost always benign course, requires an individualized approach. It seemed that when a recipient is in a life-threatening condition, we should consider taking organs from a donor suffering of papillary microcarcinoma restricted to the thyroid gland. PMID- 14529876 TI - Organ preservation solutions impair deformability of erythrocytes in vitro. AB - Microcirculatory disturbances are observed frequently after restoration of the circulation in transplanted organ. The exact mechanisms responsible for impaired microcirculation during reperfusion are unknown. Erythrocyte deformability is an important factor maintaining the microcirculation. Red blood cells (RBC) (diameter 8 microm) passing through narrow capillaries (diameter 3-5 microm) undergo deformation due to shear stress present in the microcirculation. RBC with impaired deformability may even plug the capillaries. Immediately after completion a vascular anastomosis and restoration of blood circulation in the transplanted organ, RBC may be intimately exposed to the preservation solution. The aim of the present study was to investigate the effect of the preservation solution on deformability of RBC. Blood from five healthy volunteers was withdrawn by venipuncture from an antecubital vein. EDTA was used as an anticoagulant. RBC were isolated by 10-minute centrifugation (2500 rpm) to separate serum and buffy coat. RBC washed in normal saline were resuspended in autologous plasma, Bretschneider's histidine-tryptophan-ketoglutarate (HTK) or University of Wisconsin solution (UW) at an hematocrit of 20% for incubation at room temperature. After 30 minutes RBC deformability was measured using the shear stress diffractometer Rheodyn SSD to calculate the elongation indexes of RBC at various shear stresses. HTK caused a decrease in the deformability indices of erythrocytes at shear stresses from 0.3 to 1.2 Pa. Erythrocytes incubated with UW solution revealed a decrease in deformability index at all investigated shear stresses. HTK causes less deterioration of erythrocyte deformability compared to UW solution. PMID- 14529877 TI - Impact of selected factors on early graft function in patients after renal transplantation. AB - Transplantation is the best treatment for end-stage renal diseases. For transplantologists, it is most important to know the factors that worsen graft survival prognosis. The aim of the study was to investigate factors predictive of graft loss and shortened graft survival. We retrospectively reviewed 442 renal transplant patients between 1990 and 1995 in two Szczecin units, all of whom received a triple-drug immunosuppressive regimen. One hundred thirty patients showed graft disorders such as delayed graft function or primary nonfunction. The occurrence of these disorders was examined as a function of donor and recipient age and sex, cause of ESRD, HLA compatibility, ABO and Rh compatibility, cold ischemia time, warm ischemia time, antileukocyte antibody level (PRA), and period of dialysis therapy before transplantation. The study showed that a high maximal PRA level, incompatibility for ABO group, and a longer warm ischemia time increase the probability of early graft function disorders. PMID- 14529878 TI - Influence of predialysis oxidative stress on peroxidation processes after renal transplantation. AB - Oxidative stress occurs as a result of reactive oxygen species (ROS) overproduction. The content of carbonyl groups (CG), malonyldialdehyde, 4 hydroxynonenal (MDA, 4-HNE) represent markers of protein and lipid peroxidation processes, respectively. The aim of the present study was to determine CG and MDA/4-HNE in the serum of 30 hemodialyzed patients (-HD; 13 men, 17 women of mean age 47.7 +/- 15.3 years) before and after a hemodialysis session, of 20 transplant patients (TX; 10 men, 10 women of mean age 40.7 +/- 11.3 years) before and after the procedure (RT), and of a control group (n = 47; including 30 women, 17 men of mean age 38.7 +/- 14.0 years). The CG content was evaluated using the 2,4-dinitrophenylhydrazine assay and MDA/4-HNE by the Oxis Bioxytech colorimetric method. Among the HD group the concentrations of MDA/4-HNE and CG were higher than control subjects (P <.05). In the HD group CG concentrations before and after dialysis session were similar while MDA/4-HNE concentrations were higher before the dialysis session (P <.01). One day after RT, MDA/4-HNE and CG concentrations had increased but at 7 days they had decreased and the CG level was increased. A high production of ROS can be assumed in dialysis patients. MDA/4-HNE concentrations, however, decreased after the dialysis treatment, because as low-weight molecules they diffused across the dialysis filter. On the first day after RT a high intensity of lipid and protein peroxidation was observed. During the first week after RT, accumulation of protein peroxidation products was observed but simultaneously lipid peroxidation product concentrations decreased due to quick metabolism. The intensity of lipooxidation during first day after RT seems to be dependent upon the ischemia time. PMID- 14529879 TI - Postoperative adaptation of urinary bladder to variable volume of urine in the initial period following kidney transplantation. AB - OBJECTIVES: Postoperative adaptation of urinary bladder is a process involving all patients after kidney transplantation that is facilitated by improve surgical techniques and new perioperative protocols. METHODS: The study enrolled 102 kidney transplant recipients whose transplantations were performed between 2000 and 2002. The function of urinary bladder was examined: on the day of operation and 2, 4, 8, 12, 16, and 24 weeks following operation. Patients were assessed with respect to bladder adaptation in relation to the period of dialysis. RESULTS: In patients undergoing dialysis up to 24 months, the maximum speed of flow increased from the 2nd week postoperation and regained normal values after 12 weeks. Volume of micturition, cystometric volume, and compliance regained proper values after 24 weeks of observation. CONCLUSION: The findings confirm that adaptation of the bladder is faster among patients who have had dialysis for longer than 12 months. The most useful, economical, and noninvasive assessment of bladder function was obtained with uroflowmetry in combination with daily bladder diary. PMID- 14529880 TI - 36-month follow-up of 75 renal allograft recipients treated with steroids, tacrolimus, and azathioprine or mycophenolate mofetil. AB - OBJECTIVES: The aim of this retrospective study was to assess the incidence of acute rejection episodes (AR), diabetes mellitus (DM), and serum creatinine (SCr) among renal transplant recipients treated with tacrolimus (Tac), steroids (S), and mycophenolate mofetil (MMF) or azathioprine (Aza). METHODS: Seventy-five renal allograft recipients enrolled in the COSTAMP study were followed for a period of 3 years. Patients were randomized to receive either Tac and MMF (n = 41) or Tac and Aza (n = 34) concomitantly with steroids. Follow-up assessments were performed at 3, 6, 12, 24, and 36 months. RESULTS: Patient survival at month 36 was 91.18% in the Tac/Aza/S group and 97.56% in the Tac/MMF/S group. Graft survival at month 36 was 82.35% and 85.37%, respectively. During the study period, 22 cases of biopsy-proven AR were diagnosed in 17 patients (22.6%). After 36 months the total number of AR was 11 in the Aza-treated group (32.4%) and 11 in the MMF-treated group (26.8%). DM was diagnosed de novo in 17 individuals (22.6%). During 36 months, 10 patients from Aza-treated group (29.4%) and seven from MMF-treated group developed DM (17.1%). Serum creatinine values were not significantly different in both arms of the study. Comparison of arterial blood pressure and total cholesterol revealed no significant changes in any of the studied groups. CONCLUSIONS: We conclude that combinations of steroids, tacrolimus, and azathioprine or MMF provide good results with regard to renal function. PMID- 14529881 TI - The role of the protocol biopsies in renal allograft recipients. AB - Subclinical rejection and long-term cyclosporine nephrotoxicity are well-known risk factors of chronic allograft nephropathy. In a prospective study 32 low-risk patients were randomized to either a reduced CsA dose (5 mg/kg/d) and daclizumab (group A, n = 16) for 7 months posttransplant with subsequent CsA tapering/withdrawal, or to a normal CsA dose (10 mg/kg/day) without daclizumab (group B, n = 16). Both groups received MMF and prednisone. Protocol biopsies were obtained at engraftment and 3 and 12 months after Tx. The number of rejection episodes was the primary endpoint. The secondary endpoints were: renal function, histological parameters related to CsA, and serum levels of TGF-beta and PDGF-BB. A low incidence of clinically suspected rejection episodes was observed (19% in group A and 12.4% in group B; P = NS). Although protocol biopsies showed 12 subclinical rejection episodes (six in group A, six in group B), serum creatinine levels were not different between the examined groups at 3 months. However, at 12 months, there was a statistically improved mean creatinine level in group A patients (1.2 mg/dL +/- 0.5 in group A vs 1.54 mg/dL in group B; P <.05). Chronic histopathologic changes were significant for biopsies at 3 and 12 months in both groups compared to the baseline findings for protocol biopsies (with no differences between groups, or between 3 and 12 months in both groups). Serum TGF-beta and PDGF-BB did not differ between the groups. Protocol biopsies may be useful to monitor safety and efficiency of new immunosuppressive protocols. Immunosuppressive regimens with low CsA doses followed by the drug's complete withdrawal seem to be efficient and safe in low-risk kidney allograft recipients. PMID- 14529882 TI - Biopsy eosinophilia as a predictor of renal graft dysfunction. AB - AIM: The aim of this research was to assess the impact of eosinophilia in renal biopsy specimens obtained during an acute rejection (AR) episode on the severity and reversibility of rejection and on long-term graft function. MATERIAL: Among 165 renal graft recipients who underwent transplantation (Tx) in 2001 and 2002 whose biopsy specimens revealed AR, 49 with tissue eosinophilia were compared with control group of 48 without this feature. The average biopsy time was 60.6 and 95.8 days, respectively. Biopsies during delayed graft function were performed in 46.9% of patients with eosinophilia and 29% in the control group. The immunosuppressive regimen was based on tacrolimus or cyclosporine. RESULTS: Tissue eosinophilia was observed in 49 of 165 patients (29.6%): 5 patients had eosinophilia <10/mm(2), 31 patients 10-100/mm(2), 13 patients >100mm(2) (3 patients >300/mm(2)). Severity of AR according to Banff score was statistically lower in the control group (P <.002). Patients with tissue eosinophilia who initially received steroid-free treatment presented with significantly higher (P =.02) biopsy/patients index (2.3 vs 1.81) than the total eosinophilic group. Serum creatinine values at 6 and 12 months after transplantation (Tx) were higher among eosinophilic when compared with the control group (2.41 vs 1.82 mg/dL, P <.002; 2.10 vs 1.98 mg/dL, P =.006, respectively). Chronic rejection within the first year occurred in 25% of patients with tissue eosinophilia, and 8.3% of patients in the control group. One-year graft survival rate among patients with tissue eosinophilia was lower compared with the control group (89.8% and 93.7%, respectively). CONCLUSIONS: Biopsy eosinophilia is a negative predictor that indicates a more severe course of AR and a worse response to treatment with the threat of chronic graft dysfunction. PMID- 14529883 TI - Plasma adiponectin concentration before and after successful kidney transplantation. AB - BACKGROUND: Adiponectin, a protein secreted exclusively by adipocytes, is presumed to be involved in the pathogenesis of atherosclerosis and insulin resistance. An elevated plasma adiponectin concentration was found in ESRD patients on hemodialysis (HD). However, the role of kidneys in adiponectin biodegradation/elimination is unknown. Therefore, we assessed plasma adiponectin concentrations in ESRD patients before and after successful kidney transplantation. METHODS: Among 44 hemodialyzed patients (29 men, 15 women; mean age 39 +/- 11 years; mean body mass index [BMI] 23.6 +/- 3.5 kg/m(2); mean duration of HD treatment before kidney transplantation 27 +/- 26 months), plasma adiponectin concentrations and insulin resistance indices (HOMA-R) were measured twice: immediately before kidney transplantation (Tx) and 1-2 days before patient discharge from the hospital with stable kidney transplant function (mean serum creatinine level 191 +/- 105 micromol/L). The control group consisted of 22 normotensive healthy subjects (12 men, 10 women). RESULTS: Among uremic patients, before Tx, plasma adiponectin concentrations were significantly higher than in healthy subjects (20.8 +/- 8.3 vs 8.7 +/- 4.8 microg/mL; P <.001) After successful Tx, plasma adiponectin concentrations decreased significantly (20.8 +/ 8.3 vs 15.7 +/- 7.0 microg/mL before and after Tx, respectively; P <.001). Simultaneously, after successful kidney transplantation, an increase in HOMA-R was observed (1.01 +/- 0.61 vs 1.43 +/- 0.83; P =.002). However, changes in adiponectinemia did not significantly correlate with serum creatinine or HOMA-R. CONCLUSION: The kidneys seem to play an important role in adiponectin biodegradation and/or elimination. PMID- 14529884 TI - C4d complement split product expression in chronic rejection of renal allograft. AB - Chronic allograft rejection remains the major cause of late renal graft loss. Its pathogenesis is complex, depending on both immunological and nonimmunological factors. An important role in development of chronic rejection is ascribed to an ongoing immunological reaction mainly of the humoral type. C4d complement split product, as a stable fragment of complement degradation activated by antigen antibody complexes, is considered to be an indicator of humoral activity in allografts. The aim of the present study was to establish a correlation between C4d expression and morphological findings specific for chronic rejection among biopsy specimens from patients with deteriorating graft function versus protocol biopsy specimens versus biopsy specimens of native kidneys with glomerular diseases. C4d deposits in peritubular capillaries and glomeruli were observed in 83% of patients with morphological changes of chronic rejection. No C4d expression was found in the protocol biopsy group. C4d deposits in glomeruli localizations were found in kidneys from patients with glomerulopathies; the pattern of distribution was similar to that for antibodies characteristic for glomerulonephritis. There was a positive correlation between C4d expression and morphological features of chronic rejection. In our opinion, only peritubular capillary localization is specific for a rejection process; glomerular localization is nonspecific and probably secondary to antigen-antibody complex deposition in course of some types of glomerulopathies. PMID- 14529886 TI - Polyomavirus BK infection. AB - Because it is an important factor affecting renal transplant function, BK infections are significant problem in posttransplant. BK nephropathy develops in 5% of renal allograft recipients, in most cases within the first year after the procedure. The gold standard for BK nephropathy diagnosis is still immunohistochemical staining for large T antigen in graft biopsy specimens. The aim of the present study was to evaluate the incidence of and factors influencing BK nephropathy in our renal allograft population. Among 89 renal or pancreas/kidney allograft recipients, BKV DNA was detected in 1 or more serum samples in 17 patients but BK nephropathy was diagnosed in only 1 case. Plasmacytic tubulitis was an exclusive feature in PCR-positive patients with 2 (20%) cases but no such findings in the PCR-negative group. In 40% of patients in the PCR-positive group at least 1 rejection episode was diagnosed versus 22% in the PCR-negative group. There were no significant differences in both groups according to total ischemia time, immunosuppressive treatments, or mean serum creatinine at 1 year after transplantation. PMID- 14529885 TI - Extracellular matrix proteins, proteolytic enzymes, and TGF-beta1 in the renal arterial wall of chronically rejected renal allografts. AB - Chronic rejection (CR) is the leading cause of long-term failure of transplanted kidneys. The vascular hallmark is intimal hyperplasia, accompanied by macrophage, foam cell, and T-cell infiltration. Intimal thickening results from the migration and proliferation of smooth muscle cells and increased deposits of extracellular matrix (ECM) proteins, due to release of growth factors and cytokines as well as altered ECM protein turnover. We assessed the content of fibronectin (FN) and transforming growth factor-beta1 (TGF-beta1) as well as the activities of collagenase and cathepsin B and L in renal artery walls of chronically rejected human renal allografts. We investigated renal artery samples from 8 patients with CR undergoing graftectomy, 12 patients undergoing nephrectomy, and 7 organ donors. The results were related to the DNA content of homogenates. Cathepsin B and L activities were significantly higher among those with compared with donors (P =.022). There was a trend toward higher collagenase activity in CR compared with donors and the nephrectomy group. TGF-beta1 was significantly enhanced in CR compared with donors (P =.010), and showed a trend toward higher concentrations in CR compared with the nephrectomy group. The trend was toward lower FN concentrations in CR compared with the nephrectomy group and toward higher concentrations compared with donors. Summarizing, renal CR is accompanied by enhanced proteinase activity, alterations of ECM proteins, and increased TGF beta1 in the renal artery wall. We conclude that ECM turnover and cytokines play an important role in neointimal formation and CR pathogenesis. PMID- 14529887 TI - Human herpesvirus-6 in renal transplant recipients: potential risk factors for the development of human herpesvirus-6 seroconversion. AB - Herpesviruses, including human herpesvirus-6 (HHV-6), reactivate and have the potential to be pathogenic in immunocompromised patients. Little information is available regarding the correlation between immunosuppressive therapy and HHV-6 seroconversion after organ transplantation. Serum samples obtained from 120 kidney and kidney/pancreas transplant recipients were tested to explore the potential risk factors for developing HHV-6 infection including types of immunosuppression and induction/rejection therapy. Stored serum samples obtained prior to and at the 2nd, 4th, 12th and 48th weeks after transplantation were tested for anti-HHV-6 immunoglobulin (Ig)M antibodies using indirect immunofluorescence assay. Prior to transplantation and 48 weeks after transplantation the sera were additionally tested for anti-HHV-6 IgG using enzyme linked immunoassay. Ninety-one percent of 120 recipients were HHV-6 IgG-positive before transplantation. One hundred seven of 120 patients were anti-HHV-6 IgM negative before transplantation. Primary/secondary HHV-6 seroconversion occurred in sera of 46.6% of these 107 patients. HHV-6 seroconversion most frequently occurred 2 to 4 weeks after transplantation. There was no significant relationship between HHV-6 seroconversion and the treatment with methylprednisolone (MP). The incidence of HHV-6 seroconversion was significantly higher in subjects who were treated with the regimens including Daclizumab or Sirolimus as compared with those who were on other protocols. HHV-6 seropositivity in the Polish population of organ transplant recipients is very high. We demonstrated a trend toward association of HHV-6 seroconversion with type of immunosuppressive therapy. PMID- 14529888 TI - Pneumonia in kidney allograft recipients. AB - Infectious complications, including pneumonia, remain one of the leading causes of morbidity and mortality in kidney allograft recipients. The aim of the study was to evaluate the relationship between pneumonia occurrence and treatment duration and recipient age, cause of native kidney insufficiency, dialysis duration, time between transplantation and onset, HLA matching, PRA immunosuppressive protocol, acute rejection incidence and treatment, kidney function at the pneumonia onset, as well as presence of comorbid conditions. One hundred and twenty pneumonia cases occurred in kidney allograft recipients transplanted between 1991 and 2000 with 12 to 120 months follow-up. Twenty five percentage of pneumonia episodes were diagnosed during the first posttransplant month, 25% between 2 and 6 months, and 25% at 0.5 to 3 years. Treatment duration measured from pneumonia onset to the study endpoint of recovery, which was defined as antibiotic withdrawal, show 50% of patient we cured after 15 days and 75% after 24 days of treatment. The risk of prolonged pneumonia treatment was associated with: second versus first kidney transplantation with RR = 2.3 (P <.02) and medians of treated time 28 versus 15 days; as well as serum creatinine level above 2 mg/dL (RR = 1.4; P <.098). Exposure to enhanced-potency immunosuppressive protocols including induction therapy with mono- or polyclonal antibodies increased the RR = 1.65 (P <.02), and lengthened the time to 18 versus 14 days. Maintenance immunosuppression with agents other than cyclosporine also enhanced the risk. (RR = 2.18; P <.068). PMID- 14529889 TI - Mycophenolate mofetil severely depresses antibody response to CMV infection in early posttransplant period. AB - Estimation of anti-CMV-IgG and anti-CMV-IgM is considered a relatively inexpensive screening tool of CMV status. The aim of study was to estimate how the immunosuppressive protocol influence serum anti-CMV IgG and IgM concentration in renal graft recipients and to estimate the adequacy of anti-CMV-IgG concentration and anti-CMV-IgM index as screening parameters of active CMV disease in patients receiving different immunosuppression. The study group consisted of 33 patients with clinical signs of CMV disease who received one of three types of immunosuppression: (1) azathioprine (Aza) + cyclosporine (CyA) + prednisone (Pr), 20 patients; (2) mycophenolate mofetil (MMF) + CyA + Pr, eight patients; tacrolimus (Tac) + MMF, five patients. Patients were enrolled when the pp65-antigen (pp65) of PBL was positive within 1 to 5 months after transplant (75 patients tested). The IgM-i in the Aza + CyA + Pr group was higher than in MMF + CyA + Pr group (2.73 + 1.8 vs 1.08 +/- 1.07, P =.021). The IgM-i in the Aza + CyA + Pr group was higher than in Tac + MMF (2.73 +/- 1.8 vs 0.78 +/- 0.69; P =.014). There was no difference in IgM-i between MMF + CyA + Pr and Tac + MMF. There was no difference in relative increase of IgG-c among all groups but there was a difference in relative increase of IgM-i between Aza + CyA + Pr and MMF + CyA + Pr groups (6.7 +/- 9.4 vs 2.3 +/- 5.9; P =.007) and between Aza + CyA + Pr and MMF + Tac groups (6.7 +/- 9.4 vs 0.6 +/- 0.54; P =.003). Immunosuppressive protocols including MMF exert an inhibitory influence on B-cell response and synthesis of anti-CMV-IgM. It makes the anti-CMV-IgM index an inadequate rough screening diagnostic parameter of active CMV disease. PMID- 14529890 TI - Oral cavity as a potential source of infections in recipients with diabetes mellitus. AB - Our previous observations showed alterations of oral cavity status among hemodialyzed patients and kidney allograft recipients as well as differences in the prevalence and composition of microorganisms occurring in the mouths of patients. In the present work, we analysed the results of oral cavity examinations, the identification of microorganisms, and the assessment of their importance to kidney allograft recipients or hemodialyzed patients with diabetes mellitus, in comparison with nondiabetic recipients, dialyzed patients, and control patients. PMID- 14529891 TI - Influence of hypercholesterolemia and acute graft rejection on chronic nephropathy development in renal transplant recipients. AB - Graft endothelial lesions resulting from acute rejection may be sustained by concomitant hypercholesterolemia, thus increasing the risk of chronic graft failure. The present study was undertaken to examine the influence of hypercholesterolemia and acute graft rejection (AGR) episodes on graft function and graft loss due to chronic nephropathy. A cohort of 336 patients transplanted between 1993 and 2000 having graft function at 12 months after transplantation were examined. Immunosuppressive therapy consisted of CsA, azathioprine, and corticosteroids in 90% with 10% of patients receiving mycophenolate mofetil in place of azathioprine. During the first year after transplantation, AGR occurred in 134 (39.8%) and hypercholesterolemia (6.2 mmol/L) in 132 (39.2%) of patients. The population was divided into four groups according to AGR occurrence and cholesterol concentrations during the first year after transplantation for analysis of serum creatinine concentrations and graft loss at 5 years of follow up. Patients with AGR irrespective of cholesterol levels displayed significantly higher creatinine concentrations. Graft loss in these patients increased over twofold compared to the remaining groups. Patients without hypercholesterolemia and AGR showed normal creatinine concentrations and low graft loss rates during 5 years of follow-up. PMID- 14529892 TI - Tacrolimus decreases tubular phosphate wasting in renal allograft recipients. AB - The aim of the study was to elucidate whether cyclosporine- and tacrolimus-based immunosuppression impairs tubular reabsorption of phosphate after kidney transplantation. Sixty cadaveric allograft recipients were included in the study. Forty patients receiving triple immunosupression with cyclosporine, azathioprine, and prednisone were studied for 1, 6, 12 months (groups A1 and A2, 20 patients) and for 24, 30, and 36 months (groups B1 and B2, 20 patients) after transplantation. Twenty patients who received tacrolimus with steroid withdrawal after 3 months were included in the study (group C). Recipients from groups A2 and B2 were treated additionally with vitamin D and calcium carbonate. Serum iPTH, 25-OHD, 1.25(OH)(2)D concentrations were determined, and TRP (mmol/L) and TmP/GFR (mmol/L) were calculated using Walton-Bijvoet nomogram. Higher values of total calcium serum concentration in group A were detected. Lower inorganic phosphate serum concentrations were detected in groups A and C, in contrast to group B where they remained within normal values. TmP/GFR values were significantly higher in group C in the first and third examination in comparison with patients of group A. Moreover, TRP index values were significantly higher than analogous values of groups A and B. Tacrolimus-treated patients exhibit significantly faster recovery from tubular phosphate reabsorption impairment compared to cyclosporine-treated recipients. No correlation between iPTH, 25-OHD, 1,25(OH)(2)D concentration, and tubular dysfunction parameters was observed. Amelioration of phosphate handling, in spite of hyperparathyroidism intensity, can follow early steroid avoidance. PMID- 14529893 TI - Posttransplantation diabetus mellitus under calcineurin inhibitor. AB - BACKGROUND: The development of postransplantation diabetes mellitus (PTDM) is a serious complication of kidney transplantation. PTDM has a major impact on quality of life decreasing rates of patient and graft survival. It is well known that some currently used immunosuppressants are diabetogenic. Greater diabetogenicity of FK-506 has been reported in multicenter trials. We initiated a study of conversion from tacrolimus (FK-506) to cyclosporine (CsA) among kidney allograft recipients presenting with PTDM to evaluate whether this maneuver would ameliorate a diabetic state. METHODS: This analysis of 20 adult, renal allograft recipients presenting with PTDM assumed the need for insulin therapy or oral hypoglycemics before and after conversion of the immunosuppressive regimen. The criteria for evaluating the outcome were as follows: dose reduction of insulin or oral hypoglycemic agents, adequacy of glucose control, C-peptide levels, and insulin concentration. RESULTS: During the follow-up, we observed an improvement in the control of blood glucose in the converted group. In 13 patients, satisfactory glucose control was obtained without insulin or any other agent. In 3 patients a significant dose reduction of required insulin was possible. In another 2 patients who were insulin-dependent, the switch to oral hypoglycemic treatment was clinically possible after conversion. After conversion we observed significantly lowered fasting blood glucose levels and increased C-peptide levels. CONCLUSIONS: The conversion from a tacrolimus to a CsA-based immunosuppressive regimen resulted in better glucose metabolism. We demonstrated a positive effect of conversion on the diabetic state of patients with PTDM. PMID- 14529894 TI - Thrombin-activatable fibrinolysis inhibitor in kidney transplant recipient with dyslipidemia. AB - Kidney transplant recipients are not only prone to dyslipidemia but also have a high risk of cardiovascular death. Impairment of the fibrinolytic system is thought to be one factor playing a role in development of thrombotic complications. Thrombin-activatable fibrinolysis inhibitor (TAFI) is a glycoprotein, linking coagulation and fibrinolysis. The purpose of this study was to assess TAFI concentrations and activities in renal transplant recipients stratified based upon serum cholesterol values above 220 mg/dL or below 200 mg/dL. The groups did not differ regarding age, creatinine clearance, BMI, time after transplantation, albumin, fibrinogen, thrombomodulin, or PAP. Additionally, we evaluated thrombin activity (thrombin-antithrombin complex TAT, prothrombin fragments 1 + 2); TAFI activator; thrombomodulin (TM), catalyzer of TAFI activation; and the degree of plasmin generation (plasmin-antiplasmin complex PAP) using commercially available kits. In patients with hyperlipidemia significantly higher TAFI concentrations and activities may contribute to prolonged ECLT and lowered fibrinolytic activity index (FAI). Increased levels of F1 + 2 and TAT were observed in hypercholesterolemic patients, indicating enhanced thrombin generation. Elevated TAFI concentration, and activities and enhanced thrombin generation observed in hypercholesterolemic kidney transplant recipients may contribute to hypofibrinolysis and progression of atherosclerosis in this group of patients. PMID- 14529895 TI - Is there any relation between thyroid gland function and kidney transplant function? AB - INTRODUCTION: Patients with chronic renal failure exhibit abnormalities of thyroid function. Reports regarding thyroid function in kidney transplant recipients (TX) are rare, particularly those individuals on long-term immunosuppression. The aim of this study was to investigate correlations between FT3, FT4, TSH concentrations, thyroid volume, and graft function. MATERIAL AND METHODS: The study enrolled 46 kidney allograft recipients (aged 27-67 years,) engrafted between years 1994 and 2000 and clinically stable. The mean time after TX was 45.3 +/- 37.4 months. Transplanted patients received prednisone, cyclosporine, and azathioprine. The control group included 22 patients with normal renal function. In addition to serum creatinine, TSH, FT3, and FT4 concentrations, thyroid examinations were performed with a 7.5-MHz linear probe to calculated the thyroid volume. RESULTS: Thyroid volume in TX patients was 25.3 +/- 13.3 mL. A positive correlation existed between thyroid volume and serum creatinine (P <.05), and a negative one between thyroid volume and TSH (P <.05). No correlation was observed between TSH, FT4, and serum creatinine. The time after TX was negatively related to TSH (P <.05). A negative correlation existed also between FT3 and creatinine in TX patients (P <.05). In the control group the concentrations of TSH and FT3 were within normal ranges. CONCLUSION: The FT3 concentration correlates with function of the renal graft. In TX patients the supplementary thyroid hormone therapy should be considered. PMID- 14529896 TI - Function, structure, and volume of thyroid gland following allogenic kidney transplantation. AB - The aim of the study was to assess the structure, volume, and function of the thyroid gland following kidney transplantation compared with those features of long-term transplant recipients as well as patients with normal native kidney function. Study group A consisted of 30 patients undergoing allogenic kidney transplantation, study group B included 30 long-term kidney transplant recipients who displayed stable renal function at 4 to 11 years following transplantation; control group C comprised 38 patients who were diagnosed or treated for reasons other than thyroid or renal insufficiency. Mean FT-3 concentrations in group A decreased from 2.19 pg/mL preoperatively to 1.52 pg/mL on the first posttransplantation day, returning to the preoperative values (2.06 pg/mL) at 30 days postoperatively. After 6 months the concentrations of thyroid hormones were similar to those among the long-term posttransplantation group (group B), although still lower than those in the control group. Mean thyroid volume in dialyzed patients was 17.10 mL; in the long-term group, 17.60 mL; and in the control group, 15.82 mL between groups that were not statistically significant. Abnormal structure of the thyroid gland was observed in 63% of group A (n = 19), 70% of group B (n = 21), and 29% of the control group. Significantly more abnormal thyroid gland structures were observed among dialyzed or transplanted patients. The thyroid volume was similar in all groups. Significant transient decrease in thyroid stimulating hormone (TSH) and free triidothyronine (FT-3) was not free thyroxine (FT-4) concentrations following kidney transplantation. Occasionally, increase accompanied by a change in FT-4 and TSH concentrations were observed, and antithyroid antibodies were detected only sporadically. PMID- 14529897 TI - Osteoprotegerin and its correlations with new markers of bone formation and bone resorption in kidney transplant recipients. AB - Osteoprotegerin (OPG), a natural decoy receptor for osteoclast differentiation factor, is produced by osteoblasts in response to PTH. OPG and its ligand RANKL constitute a complex mediator system involved in the regulation of bone resorption, probably playing an important role in the homeostasis of bone turnover. At present, little is known about the effects of OPG on uremic bone. Successful kidney transplantation reverses many abnormalities of bone metabolism; however, the improvement is often incomplete. The aim of the study was to assess OPG and RANKL concentrations in long-term kidney allograft recipients and their correlations with biochemical markers of bone resorption and formation. The present studies on 48 kidney transplant recipients and 25 healthy volunteers included concentrations of parathormone, osteocalcin, bone-specific alkaline phosphatase, serum CrossLaps, calcidiol, calcitriol, ICTP, PICP, tartrate resistant acid phosphatase, beta2 microglobulin, IGF-1, IFGBP-1, IGFBP-3, OPG, and RANKL using commercially available kits for measurements. Among kidney transplant recipients OPG and RANKL did not differ between transplant patients and healthy volunteers, whereas other markers of bone formation and resorption were significantly higher in the former group. OPD was related to age, time on dialysis prior transplantation, urea, platelet count, CSA dose, azathioprine dose, 25(OH)D(3), TRAP, IGF-1, IGFBP-3, whereas RANKL was related to leukocyte count, CSA concentration and dose, urine DPD, and beta2 microglobulin content. In healthy volunteers OPG correlated only with CrossLaps, whereas RANKL correlated only with osteocalcin and TRAP. Correlations between OPG, IGF system components, and some markers of bone metabolism may indicate the role of OPG/RANKL system in the pathogenesis of bone metabolism disturbances following renal transplantation. PMID- 14529898 TI - Electron paramagnetic resonance study of erythrocyte membrane fluidity in renal transplant recipients. AB - Renal transplantation (KTx) patients receiving calcineurin inhibitors cyclosporine (CsA) or tacrolimus (TAC)-are known to be at risk for the development of posttransplant hemolytic uremic syndrome. The syndrome has been reported to occur more often with CsA than TAC treatment. It has also been noted that CsA affects erythrocyte membrane fluidity. The aim of this study was to investigate whether the impairment of erythrocyte membrane fluidity is similar among patients under treatment with different calcineurin inhibitors (CsA or TAC). Venous blood was collected from 29 KTx patients and from nine healthy volunteers. To investigate the fluidity of intact erythrocyte membranes spin label electron paramagnetic resonance spectroscopy was applied. Comparison of values for controls versus CsA-treated patients demonstrated a significant decrease in membrane viscosity in the hydrophobic region of the lipid bilayer among CsA-treated patients, whereas there was no significant difference between control and patients treated with TAC. There was no significant difference in the molecular order close to the polar heads of lipid molecules among all groups. The observed changes in erythrocyte membrane fluidity among CsA-treated patients and the lack of this phenomenon in the TAC group may correlate with more frequent prevalence of hemolytic anemia among CsA than TAC-treated patients. PMID- 14529899 TI - Hemolytic anemia after renal transplantation: analysis of case reports. AB - Hemolysis after renal transplantation in some cases is clearly related to hemolytic-uremic syndrome (HUS) and usually attributed to cyclosporine (CsA) treatment. Acute hemolysis in other recipients is related to anti-erythrocyte autoantibodies. In most cases these patients have received ABO-compatible, although ABO-nonidentical, organs, mostly from O blood group donors. We report three cases of autoimmune hemolytic anemia after renal transplantation. Two patients (patients: 1 and 2; ABO-compatible, but nonidentical kidneys) suffered acute hemolysis in the third week after transplantation. One patient (patient 3: ABO-identical kidney) suffered a chronic, subclinical course of disease beginning 5 months after transplantation. The clinical picture of this disease was completely different from HUS. The existence of severe anemia (patients 1 and 2), hyperbilirubinemia (particularly high in patient 3), increased serum lactic dehydrogenase levels, and decreased serum haptoglobin in the presence of good graft function suggested an hemolytic anemia. In all patients the direct antiglobulin test was positive. The acute or chronic symptoms of hemolysis disappeared, at 2 and 5 weeks, respectively, after conversion from CsA to tacrolimus. Hemolysis in these patients probably relates to alloantibodies derived from passenger B lymphocytes transplanted with the organs. Because hemolysis has been most frequently related to CsA therapy, it is suggested that B lymphocytes proliferated and produced antibodies because CsA effects to inhibit T cell function generally spares B-cell activity. It is proposed that a subtype of B cells, which are resistant to CsA, produces anti-A and/or anti-B antibodies. Treatment with tacrolimus appears to be successful, probably due to its alternate, and likely more effective, manner of B-cell suppression. PMID- 14529900 TI - Gingival overgrowth in kidney transplant recipients treated with cyclosporine and its relationship with chronic graft nephropathy. AB - Our previous study of a group of renal transplant recipients treated with CsA showed a significantly faster development of chronic graft failure among patients with gingival hyperplasia (GH) compared to unaffected patients. The aim of the present research was to establish the impact of CsA dose and blood levels on the incidence of chronic graft nephropathy and gingival overgrowth as well as to assess risk factors for chronic graft nephropathy. The study included 64 renal graft recipients (32 patients with GH and 32 without GH) transplanted between 1989 and 1994. There were no significant differences between the pretransplant demographic and clinical data of the patients with and without GH. Patients with GH received a significantly higher total yearly dosages of CsA compared those without GH (P <.03). Serum creatinine in the first year posttransplant in patients with GH was 1.9 mg/dL versus 1.6 mg/dL in those without GH. During 9 to 14 years follow-up, end-stage renal failure due to chronic nephropathy occurred in 18 patients (56%) with GH and eight patients (25%) without GH. Ten-year renal graft survival was 35% in GH patients and 69% in patients without GH. Ten-year patient survival was 69% in the GH group and 91% in the group without GH. CsA dosage was a risk factor for GH and for graft loss, which implies a role of CsA toxic effects on the pathological mechanisms of GH and of chronic allograft nephropathy. PMID- 14529901 TI - Surgically treated early complications after kidney transplantation. AB - Early surgical complications after kidney transplantation (KTx) remain important clinical problems. The 35 patients in whom forty-six complications appeared within 1 month required surgical treatment. The causes were divided into four groups: bleeding and/or hematoma of the perigraft region (n = 22); urological complications (n = 9); simultaneous bleeding and/or hematoma and urological complications (n = 6); and others (n = 9). Among the 28 cases of hemorrhagic complication, the source of bleeding was not localized during the reoperation in 53.7% cases. Vascular anastomotic leakage was confirmed only in 7.1% of patients. The most common urological complications were stricture of (46.7% cases) and leakage at (26.7%) the vesicoureteral anastomosis. Within 3 months after KTx nephrectomy was performed in 27.5% of patients who had been previously operated for surgical complications compared to 4.6% patients without interventions. Among patients with a single reoperation the graft had to be removed in 20.0% compared with 44.4% for those with multiple reoperations. Localization of the bleeding source causing an early perigraft hematoma is not always possible. The most common early urological complication is a vesicoureteral stricture caused by edema. Surgical complications that appear within 1 month after KTx increase the risk of early graft loss. PMID- 14529902 TI - Anatomical alteration of the vascular tree observed during living related liver transplantation. AB - INTRODUCTION: The number of available cadaveric donor organs has reached a plateau. One current solution has been to increase number of living related liver transplantations. MATERIAL AND METHODS: Since October 1999 in the Department of General, Transplant and Liver Surgery, Medical University of Warsaw, 40 living related liver transplantation have been carried out. RESULTS: In 31 (77.5%) cases, a normal arterial supply was observed: the common hepatic artery arose from a celiac trunk. In two cases (5.0%), there was a partial arterial blood supply by the right accessory hepatic artery originating from the superior mesenteric artery. In two cases (5.0%), a right hepatic artery arose completely from the superior mesenteric artery (replaced artery). In one case (2.5%), a common hepatic artery originated from the superior mesenteric artery. In two cases (5.0%), an accessory left segmental artery originated from the left gastric artery. In two cases (5.0%), the function of an absent left hepatic artery was assumed by a replaced left hepatic artery originating from the left gastric artery. In two (5.0%) cases, there were two separate ducts draining the right hemiliver. There were two (5.0%) cases of an accessory duct draining segment IV, originating within the confluence of the right and left hepatic ducts. In one (2.5%) case, the common hepatic duct showed a trifurcation. CONCLUSION: During harvesting from a living donor knowledge of anatomical variants must be used to optomize the liver graft. PMID- 14529903 TI - Bile duct variations in partial liver transplantations from living-related donors. AB - The aim of this paper was to present anatomic variations of bile ducts and their effect on the perioperative course of living-related donors in partial liver transplantations in children. Liver fragments for partial transplantation were harvested from 41 related donors. Segments II and III were harvested from 35 and segments II, III, and IV from 6 donors. During the procedure, cholangiography through cystic duct was performed revealing a normal anatomy of the bile ducts in 33 (80.5%) cases. The rest of the donors showed anatomic variations. There was only one case of complications related to the bile duct. The intraoperative diagnosis of anatomic variations allowed for safe partial liver harvesting. PMID- 14529904 TI - Harvesting liver fragments from living-related donors: a single-center experience. AB - The aim of the study was to estimate the risk of harvesting a liver fragment from a living-related adult donor. Liver fragments were harvested from 44 donors. Liver segments II and III were harvested from 36 donors. Liver segments II, III, IV were harvested from 6 donors, 2 donors gave segments V, VI, VII, and VIII. After preliminary donor selection volumetric assessment of liver segments by computed tomography and arteriography was performed to visualize the cenac trunk and superior mesenteric artery. None of the donors died. No complications were observed during the operation. Only one case, a bile collection, was observed after surgery. We treated this patient with a satisfactory result by sonography guided drainage. We observed temporary elevation of bilirubin and transaminase levels and a decrease in prothrombin index value. Blood transfusion was not necessary during any of the procedures. Mean hospitalization time after the surgery was 9.4 days. Mean graft weight/recipient weight ratio was 2.54%. The risk of the harvesting liver fragment from a living-related adult donor seems to be minimal. PMID- 14529905 TI - Hepatic artery reconstruction prior to orthotopic liver transplantation. AB - BACKGROUND: This study examines the types of arterial reconstruction for grafts prepared for orthotopic transplantation procedures. METHODS: Between 1993 and February 2003, 200 organs were harvested for orthotopic liver transplantation. Arterial variations were found in 28 cases (14%), among which 16 cases (8%) required vascular reconstruction with 4 cases due to accidentally damaged during liver harvesting. RESULTS: Among the 200 organs harvested for liver transplantation, arterial variations requiring reconstruction were found in 12 cases (6%); these included: replacing an accessory left hepatic artery from the left gastric artery (9/1 reconstruction); replacing an accessory left hepatic artery from the upper mesenteric artery (2/1 reconstruction), and replacing an accessory right hepatic artery from the upper mesenteric artery (10/10 reconstructions). The splenic artery was typically used for anastomosis (seven cases, 58.3%) as well as the gastroduodenal artery (two cases, 16.7%) or the right gastric artery (one case, 8.3%). In the remaining two cases, a more complex technique was required. CONCLUSIONS: Reconstruction of graft vessels before an orthotopic liver transplantation procedure does not increase the risk of vascular complications. PMID- 14529907 TI - Human cystic and alveolar echinococcoses as indication to liver transplantation. AB - The human cystic and alveolar echinococcoses are zoonotic diseases caused by larval stages of the tapeworms Echinococcus granulosus and E multilocularis. In man the liver form of the diseases develops most frequently. Recent epidemiological data indicate that the distribution of E multilocularis in the central Europe is wider than was previously anticipated; more cases of human alveococcosis during the last years have also been noted in Poland. In the present paper we analyzed several cases of human echinococcoses from Poland with respect to diagnosis and treatment as well as the indications for liver transplantation. PMID- 14529906 TI - Biochemical assessment of the early liver graft function in relation to selected donor parameters. AB - The aim of this study was to assess the usefulness of liver grafts procured from "marginal donors." Among 62 liver transplants in 2002, almost half were harvested from donors who were not deemed acceptable by other transplant units. The authors compared the data concerning the donor's status with the function of the transplanted liver. The relations between individual parameters were estimated, as well as the differences between two groups of recipients: those who received a graft from the "poorer" donors versus those who received "better" grafts. Regardless of the relations between particular parameters a statistical analysis revealed that differences of liver function that were detected during the first 5 days after transplantation disappeared thereafter. PMID- 14529908 TI - Comparison of the results of liver transplantation for elective versus urgent indications. AB - The authors present an analysis of early and remote liver transplantation outcomes related to the presence of emergent indications among 196 of the 209 operations performed from 1989 to April 2003; namely 178 elective and 18 emergent transplantations. Perioperative mortality was 15%. The survival rate during the first 12 months was 79.8% and within 3 years 73.5% among patients operated on an elective basis (UNOS 3 and 2B). In contrast, patients with acute liver failure (UNOS 1 and 2A) showed rates of 45%, 50%, and 47%, respectively. Liver transplant outcomes depend primarily on the urgency of an operation. Longterm results are much better among patients operated on electively. Liver transplantation in patients with acute hepatic insufficiency is burdened with a high 45% mortality. PMID- 14529909 TI - Results of liver transplantation according to indications for orthotopic liver transplantation. AB - OBJECTIVE: This study assessed the results of liver transplantation in patients with a variety of different indications. METHODS: From 1989 to April 2003, 209 orthotopic liver transplantations (OLTx) were performed on 196 patients, including 178 cases. The diagnoses were: PBC (n = 34); PSC (n = 13); elective postinflammatory cirrhosis in the course of hepatitis C (n = 29); hepatitis B (n = 16); postalcoholic cirrhosis (n = 23), autoimmune cirrhosis (n = 11); Wilson's disease (n = 6); cirrhosis of unknown etiology (n = 10); secondary biliary cirrhosis (n = 5); Budd-Chiari syndrome (n = 6); and benign liver neoplasms (n = 7). RESULTS: The 3-year survival rate in the group of patients transplanted electively was 74.1%. In other groups it was: PBC, 91.4%; PSC, 69.2%; hepatitis C, 69.6%; hepatitis B, 55.5%; postalcoholic cirrhosis, 80%; autoimmune cirrhosis, 81.8%; Wilson's disease, 57.1%; secondary biliary cirrhosis, 40%; Budd-Chiari syndrome, 66.6%; hemochromatosis, 100%; benign neoplasms of the liver, 87.5%; and liver cysts, 100%. CONCLUSIONS: Results of liver transplantation were closely related to the urgency of the procedure. Better results were achieved in patients operated upon routinely compared with in those operated upon emergently (74.1% vs 50%). The best results of liver transplantation were achieved in patients transplanted on a routine basis with a diagnosis of PBC (91.4%), autoimmunologic cirrhosis (81.1%), postalcoholic cirrhosis (80%), or hemochoromatosis (100%). Patients with liver insufficiency due to hepatitis B and Wilson's disease have an increased risk of graft destruction, and the rate of survival in these patients is significantly lower than in other patients. PMID- 14529910 TI - The impact of experience of a transplantation center on the outcomes of orthotopic liver transplantation. AB - The so-called learning factor has been disregarded for many years in analyzing the causes of surgical complications and post-operative mortality; it is also the case for OLT. In our center until April 2003, 209 OLT were performed in 196 patients. We evaluated the impact of experience of the transplantation team on the outcomes of liver transplantation. Thirty-four patients died (mortality rate, 16%) and 1-year survival rate, 64%. Mortality rates varied during different periods of observation due to increasing experience of the transplantation team. The causes of mortality were assessed for a series of 34 patients: it was 75% at the beginning of transplantation procedures while recent deaths have not recently exceeded 10% of cases. PMID- 14529911 TI - Liver retransplantation in children in Poland. AB - An average of 15% of patients require retransplantation due to irreversible liver graft failure due to primary graft nonfunction, chronic rejection, vascular and biliary complications, or infections. The survival of patients and grafts after retransplantation is inferior to that after primary transplantation. The purpose of the present study was to examine the incidence, indications, and outcome of retransplantation in children. In our center 169 liver transplantations had been performed in 154 patients, and 14 patients (9%) required 15 retransplantations: nine in the early postoperative period, five late after primary transplantation, and one late after the second transplantation. One-year patient survival after primary transplantation was 82%, but after early retransplantation it was 55%. PMID- 14529912 TI - Liver transplantation across ABO blood groups in children. AB - Late results after ABOI LTx are inferior to ABO compatible organs. We report seven patients who received LTx across ABO group for emergency indications. The blood type combinations were: A to O in three, B to O in two, and B to A in two. Episodes of acute and chronic rejection, immunosuppression, and biochemical and functional tests after transplantation as well as patient and graft survival were compared between ABOI group and patients with compatible ABO group transplanted due to FLF (group I) or in an elective setting (group II). Four children are alive. Two children died of sepsis and CNS damage or MOF, and one patient died during transplantation because of cardiac failure. All recipients of ABOI grafts received immunosuppression with cyclosporine or tacrolimus and steroids. MMF was added in two subjects, and induction with antilymphocyte globulins used in five patients. An acute rejection episode was diagnosed in two recipients between 7 and 11 days after LTx. All four living patients with ABOI grafts are doing well with follow-up time between 11 months and 5 years. In one patient PTLD occurred at 1 year after ABOI LTx but was cured by discontinuation of immunosuppression and administration of rituximab. Graft survival in the ABOI group was 57.1% versus 71% in group I and 73% in group II. Respective patients survival was 57.1% 71%, and 82.0% respectively. In conclusion, in urgent cases ABOI transplantation is justified in pediatric patients when compatible grafts are not available. PMID- 14529913 TI - Liver transplantation in hepatitis C virus-related cirrhosis. AB - End-stage liver disease associated with HCV infection has become one of the leading indications for liver transplantation and it is the most common disease recurring after liver transplantation. The aim of this retrospective study was to asses factors potentially affecting outcome in patients transplanted for HCV related liver disease. Among 164 adult patients who underwent orthotopic liver transplantation from December 1994 to December 2002, 134 survived >2 months, including 25 with HCV-related liver disease. Mean follow-up after LTx was 24.8 months (range, 2.1-99.4). Anti-HCV was negative in all donors. The parameters considered in our analysis were: the course, outcome, and liver function tests at 1-year follow-up after HCV reinfection: the potential impact of maintenance and induction immunosuppressive regimens; and episodes of acute rejection. Deterioration of graft function because of HCV reinfection occurred in 16 patients (64%). Mean time for deterioration of liver function related to reinfection was 4.5 months (range, 0.83-23). Induction and maintenance immunosuppression did not affect outcome of HCV-infected liver transplant recipients. Aminotransferases were significantly higher among HCV-infected recipients than among the other patients in our series. There was a slight tendency for earlier recurrence of HCV hepatitis among patients treated with high dose steroids because of acute rejection. PMID- 14529914 TI - High-resolution 1H nuclear magnetic resonance spectroscopy analysis of bile samples obtained from a patient after orthotopic liver transplantation: new perspectives. AB - The liver is involved in many synthetic and metabolic functions. It takes part in the regulation of circulation, thermoregulation, and digestion. All liver functions are disturbed during harvesting, preservation, and transplantation. The assessment of graft function is still a matter of concern. Many methods including analysis of hepatic enzymes, ketone body ratio, and level of drug metabolites do not give complete and reliable information on graft function. Liver biopsy is still the best diagnostic tool, however, it is invasive, carrying the risk of complications. High-resolution proton spectroscopy of nuclear magnetic resonance was used for analysis of the bile sample obtained from a patient after orthotopic liver transplantation. PMID- 14529915 TI - Effect of immunosuppressive regimen on acute rejection and liver graft function. AB - Despite the use of modern immunosuppressive drugs, acute liver rejection (AR) continues to affect up to 70% of transplant recipients. The aim of this retrospective study was to assess the incidence of acute rejection episodes in patients treated with different immunosuppressive protocols. In our series, 37.3% of patients developed a clinical episode of AR. Analysis of immunosuppression has shown that the most effective immunosuppressive protocols, with regard to prevention of AR, include: antibody anti-IL-2R (anti-IL-2R) + tacrolimus (Tac) + mycophenolate mofetil (MMF) + prednisolone (Pred); anti-IL-2R + tacrolimus (Tac) + Pred; or Tac + Pred (25% vs 28.6% vs 30.4%, respectively). The highest rate of AR (66.6%) was observed among patients with anti-IL-2R and Tac but no steroid treatment, mostly (77.7%) in the initial period after liver transplantation. There were no statistical differences in liver function tests between the group treated with a CsA-based versus a Tac-based therapy. Strong immunosuppression contributed to a relatively low incidence of clinical AR in our series. The lowest rate of AR was observed among patients treated with anti-IL-2R antibody. Tac, and Pred. Deprivation of steroids in the early phase after liver transplantation substantially increased the risk of acute rejection episodes despite the use of anti-CD25. There were no statistically significant differences in liver function tests among those treated with Tac versus CsA in the short-term follow-up. PMID- 14529916 TI - Rapamycin in children after liver transplantation. AB - Experience with sirolimus (SRL) in pediatric liver transplantation (LTx) is limited. The aim of the study was to present our experience with SRL in this setting. During the last 2 years we administered SRL to 9 LTx: children in 3 due to chronic rejection and 6 due to impaired renal function. SRL was started at 2 months to 2.5 years after LTx. Target trough levels for tacrolimus for patients with chronic rejection was 8 to 10 ng/mL and SRL 10 to 12 ng/mL; for patients with impaired renal function, 4 to 6 ng/mL and 8 to 10 ng/mL respectively. For patients on only SRL and steroids the target level was 12 to 20 ng/mL. Our observation on SRL varied from 3 to 21 months, including liver function, renal function, and side effects. All patients are alive. In 3 patients with chronic rejection (ChR), follow-up biopsies showed no signs of ChR; they all normalized liver biochemistry. Independent of indication all improved their renal function. Follow-up GFR in 5 patients showed significant improvement in all. All patients showed elevated serum cholesterol values. SRL was discontinued in 3 patients due to elevation of liver enzymes in 1, persistently high serum cholesterol in 1, and repeated bouts of opportunistic infection in 1. Addition of SRL with reduced doses of tacrolimus or switching to SRL alone significantly improves renal function. PMID- 14529917 TI - Switching cyclosporine blood concentration monitoring from C0 to C2 in children late after liver transplantation. AB - BACKGROUND: Measurement of cyclospoprine (CsA) blood levels at 2 hours after oral administration (C(2)) has been proposed as a better measurement of trough level (C(0)) due to reduced intrapatient variability, acute rejection rate and renal toxicity. The aim of the present study was to assess whether there was any advantage to conversion from C(0) to C(2) CsA blood level monitoring in children late after liver transplantation. We reviewed the data from 44 children more than 1 year after liver transplantation. We measured the daily dose of CsA and the C(0) level before switching versus the daily dose and C(2) level at 6 months after conversion, in addition to the alanine aminotransferase (ALT) activity, creatinine blood concentration, and episodes of acute rejection. RESULTS: Conversion from C(0) to C(2) monitoring was not associated with a significant change in mean daily dose of CsA, mean concentration of creatinine, ALT activity or occurrence of rejection episodes. CONCLUSION: Switching from C(0) to C(2) monitoring did not seem to proffer any benefits for children late after liver transplantation. PMID- 14529918 TI - Acute liver transplant rejection: incidence and the role of high-doses steroids. AB - The aim of this study was to assess the incidence of acute rejection (AR), and the efficacy of high doses of steroids during induction of immunosuppression for AR treatment. Fifty-five patients (33.5%) experienced AR episodes in our series; but, there were no deaths or retransplantations related to AR. The median time from liver transplantation to AR was 18.5 days (range, 2-351 days). In the group with the initial dose of methylprednisolone (MP) 0.05). After 1 year observation, liver function tests were similar in both AR and non-AR groups. The only biochemical parameter that was significantly lower in the non-AR group was the aspartate aminotransferase (AST). Liver function tests determined after 1 year follow-up were not significantly different between the groups with AR treated with doses of MP lower versus higher than 1.25 g. However, liver function tests in the group treated for AR with higher doses of MP were slightly better than in the remaining subjects. Recurrence of AR occurred in 5 cases in the group with lower doses of MP (1.25 g). A relatively low dose of MP was effective to treat AR. The tendency of AR patients treated with higher dose of MP to display better liver function needs further investigation. However, AR does not seem to affect later liver function. PMID- 14529919 TI - Elevated tacrolimus levels during diarrhea in children after liver transplantation. PMID- 14529920 TI - Cytomegalovirus infection as a common complication following liver transplantation. AB - The aim of our study was to assess the incidence course, influence on liver function, diagnostic methods, prophylaxis of, and cost treatment effectiveness of CMV infection among 123 consecutive liver transplant recipients. All patients received immunoglobulin and parenterall gancyclovir as prophylaxis. CMV IgM and IgG antibodies were determined using an ELISA method. Thirty seven patients (30.0%) developed CMV infection. Main indications for primary LTX were: immune liver disease (n = 22), viral hepatitis (n = 5), and other (n = 10). CMV infection occurred between the days 5 and 416. Ten patients (27.0%) developed more than one infection (52 infections in total). Asymptomatic CMV infection was diagnosed in six cases (16.2%), CMV syndrome in 11 cases (29.7%), and hepatitis in 35 cases. All patients were treated with gancyclovir and immunoglobulin (18 cases). The intensity of infection was mild or moderate. There was no case of pneumonia or neurological disease, nor the need to use foscarnet. The correlations between the incidence of CMV infection and acute rejection, tacrolimus versus cyclosporine regimens, dual versus triple immunosupressive schemes were not statistically significant, whereas anti-IL-2R-ab antibodies markedly reduced the incidence of CMV infection (P <.05). The values of CMV IgM significantly differred before/during infection (P <.001) and before/after infection (P <.05). In conclusion, prophylaxis and antiviral treatment result in a mild or moderate intensity of CMV infection with acceptable costs. Among immunosuppressive drugs, only anti-IL-2Rab was proved to significantly reduce the incidence of CMV. PMID- 14529921 TI - Epidemiology and susceptibility to antifungal agents of fungi isolated from clinical specimens from patients hospitalized in the Department of General and Liver Surgery of the Medical University of Warsaw. AB - The aim of this study was to analyze the type and antibiotic susceptibility of fungi isolated from clinical specimens obtained from patients hospitalized in the Department of General, Transplantation and Liver Surgery of the Medical University of Warsaw between 2000 to 2002. Among the 326 clinical samples found to be positive on mycological culture, 356 strains were cultured. The most common isolates were yeastlike fungi of the genus Candida 334 (93.8%), while others included 33 other types (6.2%). The most commonly isolated species were Candida albicans, 194 strains (54.5%); Candida glabrata, 68 (19.1%); Candida krusei, 20 (5.6%); Candida inconspicua, 20 (5.6%); Candida tropicalis, 17 (4.8%); and Candida parapsilosis, 6 (1.7%). Upon testing for susceptibility to antifungal agents, all strains were susceptible to amphotericin B, while 43.8% of strains showed intermediate susceptibility to fluconazole and 25.3%, to itraconazole. Control of fungal infections in transplant and in immunocompromised patients is hindered by the low percentage of strains susceptible to commonly used antifungal agents, particularly of the triazole group. PMID- 14529922 TI - Resistance of gram-positive pathogens to antibiotics is a therapeutic challenge after liver transplantation: clinical experience in one center with linezolid. AB - BACKGROUND: Orthotopic liver transplantation has become an established therapeutic option for a large variety of fulminant and chronic liver diseases. Postoperative infections are the major cause of morbidity and the leading cause of mortality. The microbes responsible for these severe infections are predominantly gram-positive. METHODS: This article reviews results of linezolid therapy based on the clinical characteristics, microbial features, and outcomes of severe bacterial infections due to known or suspected resistant gram-positive species in selected liver allograft recipients. RESULTS: Among the 7 patients who received linezolid, methacillin-resistant Staphylococcus aureus. was isolated from 3, no pathogen from 2 patients, and serious pulmonary infection in 2 patients, 1 of whom had to be reintubated due to of respiratory failure. Cholangitis observed in 5 of 7 patients was caused by enterococci and staphylococci with septicemia in 1 subject. All patients demonstrated clinical improvement; microbiological eradication was observed in 4 patients. The majority of reported adverse events were mild or moderate in intensity. No potential drug interactions were observed between linezolid and concomitant medication. CONCLUSIONS: In the present study, linezolid proved to be effective and well tolerated. In summary, linezolid may represent an effective and safe antimicrobial agent for the treatment of infections due to susceptible and resistant gram-postive bacteria after solid organ transplantation. PMID- 14529924 TI - Anti-CD25 and tacrolimus therapy may not prevent early primary biliary cirrhosis recurrence after liver transplantation: two case reports. AB - Primary biliary cirrhosis (PBC) is an immune-mediated disorder of unknown cause characterized by progressive destruction of intrahepatic bile ducts and the presence of antimitochondrial antibodies. There is no known cure for PBC, and treatment generally includes various combinations of ursodeoxycholic acid and immunosuppressive agents. However, in most patients with end-stage PBC, liver transplantation offers a good quality of life. Recurrent PBC after transplantation is controversial, because most patients with suspected recurrent disease are asymptomatic. Antimitochondrial antibodies frequently persist and do not correlate with disease recurrence. However, most studies support disease recurrence within the graft. The effects of immunosuppression may modify or delay disease expression within the graft. If PBC recurs, intermediate-term patient and graft survivals are excellent, but the long-term outcome remains unknown. Many immunosuppressive agents have been studied with regard to their anti-recurrence properties; however, no standard therapy has been established for this group of patients. In this study we present two patients transplanted for PBC who displayed early recurrence of disease confirmed by liver biopsy and elevated serum AMA. Both individuals received the same immunosuppressive regimen. The data suggest that two doses of daclizumab and tacrolimus monotherapy in the early posttransplant period is insufficient to prevent recurrence of PBC. Addition of glucocorticoids may have beneficial effects in these patients. PMID- 14529923 TI - Renal function after liver transplantation: calcineurin inhibitor nephrotoxicity. AB - Renal failure, mainly due to calcineurin inhibitor (CNI) nephrotoxicity, is the most common complication following orthotopic liver transplantation (ltx). The aim of this study was to evaluate the incidence and course of renal failure in adult ltx patients. Severe acute renal failure in early postoperative period due to impaired hemodynamics and CNI nephrotoxicity, occurred in 14 patients, 3 of whom required dialysis. The creatinine clearance after ltx showed a tendency to decrease, but there was no statistically significant difference (P >.05) in the change in serum creatinine clearance levels between patients treated with tacrolimus (TAC) versus Cyclosporine (CsA) during the first 2 years of follow-up. Fourteen patients required conversion of their regimen because of CNI nephrotoxicity namely, dose reduction (n = 7) or discontinuation of CNI therapy with the replacement by mycophenolate mofetil (MMF) (n = 5) or SRL (n = 5). Dose reduction or CNI withdrawal significantly improved the creatinine clearance (P <.05) without affecting lives graft function. No episode of acute rejection was observed after conversion. Neither conversion of CsA to TAC nor the reverse maneuver significantly influenced the serum creatinine level (P >.05). Reduction of the CNI dose or CNI discontinuation or replacement with MMF or SRL in patients with stable liver but impaired renal function is safe, resulting in a significant improvement in renal function. PMID- 14529925 TI - Vascular complications after liver transplantation. AB - Vascular complications following liver transplantation is reviewed based upon literature data and our own results. Our study conclusions are mostly based on literature data, because our center does not have the liver transplantation experience of other centers worldwide. Thus, we may conclude, that the number and character of complications does not differ from those reported by other centers. The enbloc technique used in liver harvesting minimizes the risk of arterial damage in case of vascular anomalies. Recipient retransplantation is the most effective treatment method in cases of hepatic arterial occlusion. Doppler ultrasound examinations are effective to monitor vascular blood flow in the transplanted liver. PMID- 14529926 TI - Biliary tract complications following liver transplantation. AB - INTRODUCTION: Biliary tract complications, which occur in 5.8% to 24.5% of adult liver transplant recipients, remain one of the most common problems following transplantation. The aim of this study was to evaluate these problems and analyze methods of treatment. MATERIAL AND METHODS: From 1989 to 2003, 36 (18.7%) among 193 patients who underwent orthotopic liver transplantations in our center developed biliary complications. Biliary strictures that developed in 18 cases (9.3%) were the most common complications. Clinical manifestations of strictures developed at 2 to 24 months after transplantation. Bile leaks occurred in 10 patients (5.2%), and were diagnosed in along the T-tube 4 cases and was not accompanied by any clinical manifestation. Bile leak to the peritoneum after T tube removal occurred in 2 patients (1.1%). Solitary gallstone formation in one case (0.5%) was removed with the use of ECPW. One patient required retransplantation within 3 months after transplantation, because of the most severe complication-ischemic necrosis of biliary tract. RESULTS: Uneventful recovery was achieved in 34 patients in the analyzed group (94.4%). There was no case of recurrence during outpatient follow up. Two patients died in late follow up of unrelated causes: namely, gastrointestinal bleeding due to a duodenal ulcer and multi-organ failure (MOF) due to a third severe episode of acute liver transplant rejection. CONCLUSIONS: Biliary complications remain an important problem in liver transplantation. Endoscopic and radiologic management are effective in the majority of cases. Surgical intervention is obligatory in selected cases. PMID- 14529927 TI - Acute coagulopathy after reperfusion of the liver graft in children correction with recombinant activated factor VII. AB - BACKGROUND: Several studies have proven that massive blood loss increases postoperative morbidity and mortality in liver graft recipients. Since we have successfully corrected coagulopathy preoperatively using an intravenous (IV) bolus of recombinant activated factor VII (rFVIIa) in 2 patients with fulminant liver failure, we observed that there was rapid reversal of preexisting advanced coagulopathy in another 40 patients with high risk for intraoperative bleeding by this treatment immediately before transplantation. Recently to control hemostasis we have administered rFVIIa also to patients presenting with acute coagulopathy and nonsurgical bleeding after graft reperfusion as described herein. MATERIALS AND METHODS: We have used rFVIIa in 7 children presenting with severe coagulopathy and nonsurgical bleeding after liver graft reperfusion. The dosage of rFVIIa ranged between 37 and 148 mcg/kg. An antifibrinolytic agent (aprotinin, tranexamic acid) was administered simultaneously. RESULTS: APTT before rFVIIa was 86.10 to 183 seconds, (mean, 132.1 +/- 39.88), after the bolus of rFVIIa 49.4 to 206.1 (mean, 112.7 +/- 58.53), and at the end of surgery 71.70 to 180 (mean, 110.3 +/- 40.98). INR after reperfusion was 1.82 to 3.91 (mean, 2.56 +/- 0.67), 1.03 to 1.92 (mean, 1.54 +/- 0.35) after rFVIIa, and 1.74 to 5.58 (mean, 2.64 +/- 1.35) at the end of surgery. Before rFVIIa administration intraoperative blood transfusions after graft reperfusion were 900 to 4200 mL of red blood cells (RBC) (0.82-5.4 total blood volume) and after reperfusion 0 to 1800 mL of RBC (0-2.5 TBV). No postoperative vascular complications were observed. CONCLUSIONS: A single dose of rFVIIa effectively reverses the severe coagulopathy developing after graft reperfusion, establishing effective hemostasis in liver transplant recipients without an increased risk of thrombotic complications. PMID- 14529928 TI - Plasmacytic hyperplasia--the early form of posttransplant lymphoproliferative disorder--with atypical morphology and clinical course in patient after liver transplantation: a case report. AB - This case report describes an early lesion of posttransplant lymphoproliferative disorder (PLTD)--plasmacytic hyperplasia with atypical morphology. The 54-year old patient was 4 months after liver transplantation due to alcoholic cirrhosis. The postoperative course had been uneventful without graft rejection episodes. Primary immunosuppressive therapy included tacrolimus and prednisone. On admission to the hospital the patient showed rapidly increasing jaundice, hepatomegaly, anemia, thrombocytopenia, and significant leukocytosis. A biopsy suggested generalized infection. Acute Epstein-Barr virus (EBV) infection was confirmed using serological methods. Despite treatment the patient died. On autopsy we found features of generalized infection. Histological examination of the enlarged lymph nodes showed plasmacytic hyperplasia despite lymph node atrophy. Plasmacytic hyperplasia, an early lesion of PTLD despite usually a good prognosis with multifactor therapy may display a rapid course that leads to death through intensified immunosuppression. In accordance with other reports we confirmed reactivation of EBV infection as the probable cause of plasmacytic hyperplasia. The lymph node morphology of plasmacytic hyperplasia may be atypical with atrophy of lymphoid components accompanying plasma cell proliferation. PMID- 14529929 TI - Rapid correction of prothrombin time after low-dose recombinant factor VIIA in patients undergoing orthotopic liver transplantation. AB - Orthotopic liver transplantation (OLTx) is associated with a major risk of blood loss resulting from portal hypertension, collateral circulation, and clotting disturbances. Application of a recombinant factor VIIa (rFVIIa) has been reported to promptly correct clotting abnormalities reducing the risk of intraoperative bleeding. This study included 8 patients who underwent OLTx for end-stage liver cirrhosis, with protrombin times (PT) exceeding the upper limit of normal by more than 4 seconds before surgery. All subjects were administered a small single intravenous dose of rFVIIa [mean 68.37 microg/kg body mass (range, 32.88-71.64)] 10 minutes prior to the skin incision. The PT was then measured 15 minutes later, following graft reperfusion, and 12 hours since drug application. All patients showed rapid correction of PT within 15 minutes after injection (median PT before injection 20.25 seconds vs 11.5 seconds after injection, P <.0001). Following the reperfusion PT was found to be prolonged again. These values are not significantly differ from those before surgery and are comparable to PT values after reperfusion in patients who did not receive rFVIIa. None of the patients developed thromboembolic complications. In conclusion, lower than recommended dose of rFVIIa caused rapid improvement in the PT shortly after injection. After reperfusion PT became prolonged again, which may account for the lack of thromboembolic complications observed in this group of patients. PMID- 14529931 TI - Deterioration of renal function after replacement of cyclosporine with sirolimus in five patients with severe renal impairment late after heart transplantation. AB - A retrospective analysis of all cases when cyclosporine (CyA) was replaced with sirolimus (SIR) to avoid the renal toxicity of CyA late after heart transplant (OHT) was discontinued due to advanced renal impairment in all five heart transplant recipients (four men, 1 women; age 41 years, range 38-45; time after OHT 5 years, range 4-14). The serum creatinine level at the time of SIR introduction, which was 298 micromol/L (range 217-676), had remained stable for the 6 months prior to conversion. Target SIR trough levels were 12-20 ng/mL. In four patients the last dose of CyA was immediately followed by the first dose of SIR, whereas in one patient CyA was tapered gradually in the presence of low-dose SIR. Deterioration of renal function with signs of fluid overload and increased serum creatinine levels (Delta: 77, 33-150 micromol/L) was observed in all patients. Two patients required dialysis during SIR treatment including one case of pulmonary edema requiring emergency hemodialysis. None of four biopsies showed significant rejection. Four patients were converted back to low-dose CyA (including the two patients requiring dialysis during SIR therapy); one was maintained on mycophenolate mofetil. The creatinine level at the time of SIR discontinuation was (range 250-753) micromol/L, 448. Eventually, all patients required dialysis. In conclusion, replacement of cyclosporine with sirolimus in heart transplant recipients with severe renal impairment late after transplantation may accelerate renal failure. PMID- 14529930 TI - Pathologic assessment of cardiomyocytes in heart transplant recipients treated with rapamycin or cyclosporine. AB - The aim of this study was to compare cardiomyocytes and stromal pathology in heart transplant recipients treated with rapamycin (RAPA) versus cyclosporine (CyA). We analyzed elective biopsies obtained during first 3 months after heart transplantation in four patients treated with RAPA (24 biopsies) and seven patients receiving CyA (49 biopsies). Additional medications in both groups consisted of mycophenolate mofetil or azathioprine and prednisone. The intensity of rejection was assessed using the ISHLT scale; it was comparable in both groups based upon the number of results showing significant rejection and the average biopsy scores. Each slide was also examined under high-power magnification to sarcoplasmic and nuclear changes. Sarcoplasmic vacuolation, premyocytolysis and myocytolysis, nuclear staining, stromal fibrosis and edema, presence of vasculopathy, and lymphocytes infiltrating the myocardium occurred more frequently in the CyA group. The difference in the degree of hyperchromasia of the nuclei was highly significant (67% versus 10%, P <.00001). Our findings suggest that despite comparable levels of rejection as assessed using the ISHLT scale, patients treated with RAPA display fewer signs of cardiomyocytic alterations early after heart transplantation. PMID- 14529932 TI - Limited utility of cyclosporine C2 monitoring in heart transplant recipients receiving ketoconazole. AB - The aim of the study was to compare 2 hours postdose concentration (C2) of CyA in stable patients receiving ketoconazole concomitantly late after heart transplantation (OHT) with patients not receiving ketoconazole. Routine C2 and C1 (1 hour postdose concentration) of CyA monitoring (FPIA, AxSYM, Abbott) along with C0 (trough level) were performed in 64 elective patients. The KETO group consisted of 29 patients receiving 200 mg of ketoconazole daily along with CyA; the remaining 35 patients were included into the control group. Patient characteristics (KETO vs control group) were as follows: age, 49 +/- 11 versus 48 +/- 12 years; percentage of male patients, 93 versus 80; follow-up post-OHT, 4.3 +/- 2 versus 5.3 +/- 2 years. Target C0 of CyA was 175 to 225 ng/mL; CyA doses remained stable for at least 1 month. We compared maintenance doses of CyA, C0, C1, C2 of CyA, number of biopsy-proven acute cellular rejection (AR) during the one year and after the first year post-OHT, and creatinine in both groups. Statistical significance was assessed using Mann-Whitney U test. Results were as follows (KETO versus control group): CyA dose, 53 +/- 30 versus 216 +/- 69 mg, P <.000001; C0, 181 +/- 77 versus 160 +/- 53 ng/mL, NS; C1, 406 +/- 78 versus 803 +/- 317 ng/mL, P =.000001); C2, 397 +/- 174 versus 689 +/- 284 ng/mL, P =.000001, AR during the first year after OHT, 2.8 +/- 1.9 versus 2.3 +/- 1.6, NS; AR beyond first year after OHT, 0.2 +/- 0.5 versus 0.7 +/- 0.9, P =.03); creatinine, 181 +/ 50 versus 160 +/- 114 micromol/L NS. In conclusion; C2 monitoring in stable heart transplant recipients receiving cyclosporine and ketoconazole concomitantly late after procedure does not seem to be sufficient to estimate the immunosuppressive effect of this combination. PMID- 14529933 TI - Simultaneous pancreas-kidney transplantation: analysis of donor factors. AB - There are no urgent indications for simultaneous pancreas-kidney transplantation. So our policy is to harvest only a pancreas in good biologic condition. The criteria for acceptance of a pancreas donor are: age 15 to 40 years, ICU stay < 7 days, no clinical signs of infection, negative virologic status, no history of hypotension or cardiac arrest, serum amylase elevation below three times normal values, controllable hyperglycemia, no history of pancreatic disease, no history of abdominal trauma damaging the organ, no history of alcohol addiction, BMI < 25, no functional or anatomical lesions of the kidneys, and expected ischemia time less than 12 hours. The proper selection of a pancreas donor allows one to achieve good insulin secretion immediately after transplantation. In 2000 to 2002 all 20 pancreases transplanted at transplant center displayed immediate secretory function after transplantation. PMID- 14529934 TI - Do high levels of serum triglycerides in pancreas graft recipients before transplantation promote graft pancreatitis? AB - OBJECTIVE: Graft pancreatitis is a serious complication following pancreas transplantation. The aim of this study was to evaluate the influence of pretransplant serum lipid levels on the development of graft pancreatitis among patients undergoing simultaneous pancreas and kidney transplantation (spkTx). METHODS: We reviewed data from spkTx patients engrafted between 1999 and 2002. Group 1 consisted of 10 recipients with well-established pancreas and kidney graft function without postoperative pancreatitis; group 2 5 spkTx recipients who developed fatal graft pancreatitis in the first posttransplant month. The lipid parameters evaluated within 1 hour before transplantation and after hemodialysis included total cholesterol, HDL, LDL, VLDL, triglicerides and apoproteins A and B. RESULTS: Triglycerides, apoprotein B and VLDL were significantly increased just before transplantation among patients who developed fatal pancreatitis compared to those patients with good graft function. CONCLUSION: Recipient hypertriglyceridemia promotes graft pancreatitis in previously injured pancreatic graft. PMID- 14529935 TI - Proliferation of transplanted allogeneic pancreatic islets. AB - Recent studies showed enhanced regeneration of pancreatic islets in some circumstances. The purpose of our study was to investigate the proliferate potential of rat pancreatic islet cells in allogeneic grafts. Adult Lewis female rats and WAG male rats served as recipients and donors, respectively. Diabetes was induced by single intravenous (IV) injection of streptozotocin producing diabetes as confirmed by nonfasting plasma glucose >300 mg% on 3 consecutive days. Islet rejection was considered complete when glycemia exceeded 250 mg% and was confirmed by histopathological examination. To obtain long survival of allogeneic islets a tolerance-inducing method used allogeneic UV-B irradiated bone marrow transplantation into nonlethally selectively cytoreducted recipients with a donor-type splenocyte infusion followed by cyclophosphamide 200 mg/kg bw. Endocrine cell proliferation was assessed morphometrically using double immunostaining for pKi-67 and insulin or glucagon. Double immunolabelling, propidium iodide staining, and TUNEL assay were used to identify both proliferating and apoptotic cells. The rise of glycemia >350 mg/dL after graftectomy in euglycemic recipients was correlated with immunohistological examination, showing that the euglycemia was due to properly functioning pancreatic islet allotransplants. The immunohistochemical examination confirmed the presence of endocrine beta and alpha cells. In comparison with normal pancreas which showed 0.4 +/- 0.12%, pKi-67-positive cells, long-surviving grafts had a significantly higher proliferation capacity (5.61 +/- 0.94%; P <.001). In contrast, rejected grafts/control groups did not show significantly enhanced proliferation (0.73 +/- 0.19%), and had endocrine cells undergoing apoptosis. The incidence of apoptosis in endocrine cells within long-surviving graft appeared to be extremely low. In conclusion, the growth and death of endocrine cells in allogeneic grafts differ between accepted and rejected cases. The level of proliferation in the graft at day 150 was significantly higher compared with normal pancreatic beta cells. PMID- 14529936 TI - Assessment of some porcine strains as donors of islets of Langerhans. AB - Mass isolation of viable porcine islets is a difficult task because of their fragility, and because of donor variability with respect to strain, age, sex, feeding, and methods of slaughtering. Not all strains are equally suitable for islet separation. The aim of this study was to evaluate porcine pancreata as an alternative source of islets for clinical transplantation. Pancreata were digested from pig strains available in Poland: 248 market weight slaughterhouse pigs and 42 pigs, belonging to the Polish Large White (WBP, 14 sows and 3 males), Polish White Pendant-Ears (PBZ; 16 sows), Pietrain (8 sows), and Yorkshire (1 sow) races. Prepurification data of recoverable islets/g and islet equivalents/g were considered as representative for the number of recoverable islets. Acceptable results namely, islet and/or islet-equivalent (IE) number of at least 1000/g, were obtained from only 56 of 248 slaughterhouse pigs, namely 2073 +/- 137.4 SE (median 1767/g) islets with values of IE of 2994 +/- 303 SE (median 1874/g). Our data support Krickhahn et al suggesting that only pancreata with an average islet size exceeding 199 microm should be digested and that only from 1 of 3 to 5 porcine pancreata is an adequate amount of islets generated. PMID- 14529937 TI - Amylase levels in preservation solutions as a marker of exocrine tissue injury and as a prognostic factor for pancreatic islet isolation. AB - Occurrence of primary graft nonfunction of pancreatic islets demands research for new methods of organ preservation during cold ischemia conditions. Digestive enzymes released during preservation injure the islets for subsequent rewarming and islet isolation processes. The aim of our study was to assess the amylase level in preservation solution as a marker of exocrine tissue injury, allowing the prognosis of islet yield and viability. The experiments undertaken on rats used three commercially available preservation solutions: ViaSpan (UW); Custodiol (HTK); and Euro-Collins (EC). After 180 minutes of cold ischemia, the highest islet recovery was observed among pancreata stored in UW solution (508 +/- 139 vs HTK 344 +/- 103; P <.05 vs EC 322 +/- 113; P <.05). These islets also revealed the highest insulin stimulation index in glucose static tests (1.19 +/- 0.30 vs HTK, 0.87 +/- 0.43; P <.01, vs EC.25 +/-.06; P <.001). The highest amylase level in the preservation solution was associated with a decreased yield of islets during the isolation process and lowest insulin stimulation index (increasing 139 +/- 18% for EC, 108 +/- 12% for HTK; P <.05 vs 87 +/- 10% for UW; P <.05). Our data strongly suggest, that the dynamic of amylase release during pancreas preservation at 4 degrees C correlates with a reduced number and viability of isolated islets. These results suggest that measurement of amylase levels after pancreas preservation may have potential clinical application as a marker to evaluate pancreatic tissue injury. PMID- 14529938 TI - Evaluation of relative anti-myeloma activity of high-dose melphalan followed by the first peripheral blood stem cell transplantation, as compared with the second transplantation, and to VAD chemotherapy. AB - Current treatment in multiple myeloma consists of three courses of chemotherapy in low doses with subsequent hematopoietic stem cell mobilization to the peripheral blood using high-dose cyclophosphamide, collection and conditioning with high-dose chemotherapy (melphalan) followed by retransplantation of autologous peripheral blood stem cells (PBSCT). Only a few studies compare the effects of different phases of therapy on parameters, such as monoclonal immunoglobulin level and the presence of malignant CD38(+) and CD56(+) cells in blood and marrow. The aim of this study was to compare the effects of these two major phases of treatment (conventional and high dose) in the same patients, and furthermore, to compare the effects of the second course of high-dose therapy followed by PBSCT with the effects of the first one. Fifteen patients were included in the study. On average, conventional chemotherapy only slightly reduced the values of all disease markers. In contrast, high-dose therapy resulted in a dramatic effect, rapidly normalizing the values of all parameters. The effects of second PBSCT were only modest compared to the first. These data suggest that high-dose therapy is an efficient method to reduce tumor load in multiple myeloma. Conventional-dose chemotherapy may be simply a waste of time for some patients and may be either omitted or administered after high-dose therapy to consolidate remission. PMID- 14529939 TI - Triple transplantation of autologous peripheral blood stem cells each time following conditioning with 100 mg/m2 of melphalan for multiple myeloma patients in poor performance status. AB - Approximately one third of multiple myeloma patients (below 60 years) are diagnosed either in advanced disease or with significant comorbidities. Many other patients referred to transplant centers have already been heavily pretreated with multiple courses of various conventional chemotherapies. These patients are frequently in bad or even grave clinical condition; they are unlikely to survive standard high-dose melphalan (200 mg/m(3)) chemotherapy and autologous hematopoietic stem cell transplantation. Palumbo et al reported a protocol for elderly patients that utilized reduced conditioning (melphalan 100 mg/m(2) three times at 2-month intervals, each time supported by autologous hematopoietic rescue). We have used this protocol as a start to develop a method to induce a remission in the aforementioned subgroup of myeloma patients. Patients with stage III disease and WHO performance status 2 or higher are treated with one or two cycles of cyclophosphamide (2 to 4 g/m(2)) and undergo peripheral blood stem cells collection. Subsequently, they are treated with three to four doses of melphalan (100 mg/m(2)) at 8- to 12-weeks intervals each time supported by infusion of peripheral blood stem cells. To date 13 patients have been entered into the protocol. With one exception of transiently stable disease, the remaining patients obtained at least partial remission and three, complete remission. The compliance was good and better with each subsequent course. For half of the patients the problem was a short duration of response. This method when developed may offer a new treatment alternative for a subgroup of high-risk multiple myeloma patients. PMID- 14529940 TI - Clinical and experimental transplantation of isolated organ or tissue cells indications problems from the Polish perspective. AB - Clinical cell transplantation remains as a clinical experiment. Morphologically intact and functional cells transplanted into their tissue of origin undergo rapid disintegration by attacking granulocytes and macrophages, leading to the so called early graft dysfunction. When transplanted to sites remote from their origin, the process of elimination is even faster. Among millions of transplanted cells, only a few survive in an autologous or syngeneic recipient, not to mention an allogeneic combination. What is the mechanism of elimination of the majority of transplanted cells? Which cells survive and what is their genotype and phenotype? There are several problems that should be investigated: (1) the mechanism of anoikis, namely, detachment-induced apoptosis, (2) the reaction of the innate immune system to transplanted cells, (3) the microchimerism created by the transplanted cells, and (4) the "niche" for transplanted cells within the local cell chemical environment and signalling. Our experience with transplantation of hepatocytes illustrates current problems in isolated cell grafting. Proposals to increase the in vivo survival rate of transplanted syngeneic hepatocytes are essential to the vision of the future of cell transplantation. PMID- 14529941 TI - Transplantation of hepatocytes: elimination of recipient natural killer cells with irradiation and bone marrow reconstitution prevent early graft dysfunction. AB - Transplanted isolated syngeneic and allogeneic hepatocytes rapidly disintegrate, irrespective of the origin or the site of engraftment namely spleen, liver, portal vein, peritoneum, or subcutaneous tissues. Although various methods have been applied to attenuate this reaction, none have been found effective. We applied a combined protocol consisting of administration of anti-asialoGM1 antiserum (eliminating NK cells), sublethal whole-body irradiation, and reconstitution with syngeneic bone marrow cells to intrasplenic hepatocyte transplantation and 3 consecutive partial hepatectomies. This method overcame the early disintegration of grafted hepatocytes. Ninety days after transplantation numerous hepatocyte clusters and dilated bile canaliculae, occupying two thirds of the spleen, were observed, with some hepatocytes adhering to the bile ducts forming Hering's canals. Mitotic figures were noticed. There were no recipient mononuclear infiltrates around the hepatocyte clusters. PMID- 14529942 TI - The effect of cyclosporine and tacrolimus on indigenous bacterial flora in human skin grafts. AB - Allogeneic skin transplants require intensive immunosuppressive therapy. Treatment protocols used for parenchymal organ grafts are not satisfactory to prevent skin graft rejection. Another factor responsible for the destruction of allogeneic skin transplants is bacterial inflammation. Temporary ischemia and the allogeneic reaction in transplanted skin cause increased permeability of the epidermis and the dermal capillaries, making skin grafts vulnerable to bacterial penetration. Moreover, immunosuppressive therapy compromises the host immune response. The present study assessed the effects of immunosuppression by cyclosporine (CsA) or tacrolimus (Tac) on the indigenous bacterial flora of transplanted human skin. We found that a 6-day course of treatment with CsA or Tac was followed by an increased prevalence of bacterial isolates, mostly evidenced by a change in the spectrum of graft bacterial flora from Staphylococcus aureus and coagulase-negative staphylococci toward more pathogenic strains such as Escherichia coli, Enterococcus faecium, micrococcus, and pseudomonas. The mouse skin adjacent to the graft remained sterile, precluding the possibility of graft contamination with mouse flora. PMID- 14529943 TI - The emerging matrix of immunosuppressive agents. AB - In recent years, significant milestones have been reached in the field of transplantation through the development of immunosuppressive drugs that inhibit lymphocyte activation, cytokine signal transduction, and cellular proliferation. However, the widespread tissue distribution of the molecular targets exploited to date-calcineurin, mammalian target of rapamycin (mTOR), and inosine monophosphate dehydrogenase-produces an array of collateral toxicities. Avoiding these side effects requires new strategies that selectively block destructive immune responses: a fifth generation of immunosuppressants. These agents must target molecules that are critical for and specific to the adaptive immune response. PMID- 14529944 TI - The influence of calcineurin inhibitors on mycophenolic acid pharmacokinetics. AB - Despite the fact that concentrations of mycophenolic acid (MPA) are not routinely measured, accumulating data suggest the usefulness of this monitoring to optimize therapy. The aim of this study was to assess the influence of CsA and tacrolimus on MPA pharmacokinetics. Concentrations of MPA were measured using HPLC. An assay was performed before dose (the C(0)), as well as at 40 minutes and 1, 2, 4, 6, 8, 10, 12 hours after administration of mycophenolate mofetil (MMF). MPA profiles were assessed in 51 patients receiving tacrolimus at a dose of 1.0 g/d and prednisone as well as in 97 patients receiving CsA (2.0 g/d) and prednisone. Significant correlations of MPA levels with serum albumin and GFR were observed in both groups. Women presented with higher levels of MPA than men. C(0) MPA level among the tacrolimus group were significantly higher than those in CsA group: 3.18 +/- 2.21 microg/mL versus 1.68 +/- 1.03 microg/mL (P 14, 14, and 4.9, respectively) in the 6 Hz model. The results from the present study support the continued search for glutamate receptor modulators as potential antiepileptic agents. Furthermore these results illustrate the importance of using several different animal seizure models in the search for novel AEDs and the potential utility of the 6 Hz seizure model in identifying novel AEDs. PMID- 14529951 TI - Isobolographic and subthreshold methods in the detection of interactions between oxcarbazepine and conventional antiepileptics--a comparative study. AB - Until now, a character of interactions among the antiepileptic drugs (AEDs), in some experimental models of epilepsy, has been determined alternatively with subthreshold and isobolographic methods. In order to elicit the precise and adequate method for evaluating two drug interactions, the comparative study was performed in the maximal electroshock-induced seizure test in mice. In this experimental model, the exact types of interactions among oxcarbazepine (OXC) and conventional AEDs (diphenylhydantoin, phenobarbital, valproate, carbamazepine, and clonazepam) were determined with both methods. Results from the subthreshold method showed a considerable reduction of ED(50) values of clonazepam, diphenylhydantoin and valproate (after administration of OXC at the highest subthreshold dose of 2.5 mg/kg), whilst ED(50)s of carbamazepine or phenobarbital were almost unchanged when OXC (2.5 mg/kg) was co-administered with these AEDs. Results from the 2-dimensional (2-D) isobolographic analysis of interactions for a 50% anticonvulsant effect, for three fixed drug dose ratio combinations of 1:2, 1:1, and 2:1, indicate antagonism between OXC and diphenylhydantoin as regards their anticonvulsant (protective) activity. Furthermore, the interactions between OXC and clonazepam occurred either antagonistic (for the fixed-ratios of 1:4 and 1:3) or synergistic (for the fixed-ratio combinations of 1:1 and 2:1) depending on the proportions of used drugs. Remaining interactions between OXC and carbamazepine, OXC and valproate, or OXC and phenobarbital (for the fixed-ratios of 1:3, 1:1, and 3:1) were isobolographically additive for a 50% anticonvulsant effect tested. The 3-dimensional (3-D) isobolographic analysis of interactions between OXC and CZP revealed that the dual character of interactions (antagonistic and synergistic) observed for a 50% anticonvulsant effect (ED(50)) was also present for additional drug-dose effects tested, i.e. ED(16) and ED(84). The 3-D isobologram for the combination of OXC with CZP clearly visualized either synergy or antagonism between the drugs in combinations.Distinct differences resulting from two experimental methods prove evidently the superiority of isobolographic analysis over the subthreshold method. The former clearly and adequately detects the exact types of interactions between two AEDs, becoming a potent and powerful paradigm for further studies evaluating the character of interactions among AEDs. PMID- 14529952 TI - Sleep but not hyperventilation increases the sensitivity of the EEG in patients with temporal lobe epilepsy. AB - PURPOSE: To evaluate the relative impact of 3 and 5 min of hyperventilation (HV) and different sleep stages on the sensitivity of the interictal EEG in focal epilepsy. METHODS: We examined 20 patients with temporal lobe epilepsy (TLE, 85%) or extratemporal epilepsy during EEG-monitoring. We compared 6 min EEG (12 epochs of 30s) during/after each: (a) waking; (b) 5 or 3 min of HV; (c) sleep stages 1, 2, 3/4 and REM regarding the frequency of epileptiform discharges (ED). The Wilcoxon matched pairs signed rank test was used. The main endpoint was the comparison of 5 min of HV with sleep stage 2. RESULTS: During sleep stage 2, ED were more frequent than during/after 5 min of HV (P=0.002). Compared to the waking EEG, all NREM-sleep stages activated ED. Sleep stage 2 was associated with the strongest activation. There was no difference between the waking state and REM-sleep. Compared to the waking EEG, neither 3 nor 5 min of HV showed an activation of ED. CONCLUSION: In patients with TLE, sleep stage 2 shows a significantly higher sensitivity for ED than 5 min of HV. Compared to the waking EEG, HV showed no activating effect on ED. These results suggest that in patients with the clinical diagnosis of TLE (and possibly other focal epilepsies) measures to record sleep stage 2 (such as sleep deprivation) should be increased whereas HV appears to be dispensable in this setting. PMID- 14529953 TI - Regulation of epileptiform activity in hippocampus by nicotinic acetylcholine receptor activation. AB - Nicotinic acetylcholine receptors (nAChRs) regulate neuronal excitability within the CNS. To assess the possible modulatory influence of nAChRs on epileptiform activity, a range of nAChR ligands were applied during experimentally induced epileptiform activity in rat hippocampal slices. Bath application of the potassium channel blocker 4-aminopyridine (4AP; 10-50 microM) resulted in the development of spontaneous epileptiform bursting activity in area CA3 that consisted of short duration (257+/-15 ms) field events occurring regularly at a frequency of 0.4+/-0.02 Hz. Subsequent co-application of the selective nAChR agonists 1,1-dimethyl-4-phenyl-piperazinium iodide (DMPP; 0.3-300 microM), choline (0.01-3mM) and lobeline (3-30 microM) produced sustained and concentration-dependent increases in burst frequency with maximal frequency potentiation of 37+/-5%, 27+/-5% and 24+/-11%, respectively. DMPP (10-30 microM; n=31) also potentiated epileptiform bursting induced by reducing GABA(A) receptor mediated synaptic transmission using 20 microM bicuculline or enhancing NMDA receptor-mediated excitation by lowering extracellular Mg(2+). Irrespective of the epileptiform model studied all nAChR agonist induced frequency potentiation was reversed upon washout of the agonist or co-application of one of the selective nAChR antagonists dihydro-beta-erythroidine (10-30 microM), mecamylamine (50-200 microM) or alpha-bungarotoxin (100 nM). These results provide compelling evidence that activation of nAChRs exacerbate epileptiform activity in the rat hippocampus. PMID- 14529954 TI - Relationship between lamotrigine oral dose, serum level and its inhibitory effect on CNS: insights from transcranial magnetic stimulation. AB - The antiepileptic drug lamotrigine (LTG) is known to reduce cortical excitability evaluated by transcranial magnetic stimulation (TMS). We investigated the relationship between LTG oral dosages, serum levels and inhibitory effects on resting motor threshold (RMT), a parameter of motor system excitability assessed by TMS. In a randomized, placebo-controlled crossover study 16 male volunteers received 325 mg LTG as a single dose, as bi-hourly graded cumulative dose, or placebo. RMT and serum levels were measured before and after 2-8 h. With single dose, RMT elevation showed a poor but significant correlation to serum levels. With graded dose, serum levels as well as RMT increased dose-dependently with significant (P<0.0001) linear correlation. However, detailed comparison showed a high inter-individual variability in the relationship resembling a sigmoid correlation. Different mechanisms besides the sodium-channel blockage as the main mode of action of LTG are discussed to explain the diversity of individual dose response relationships. Provided that the RMT elevation reflects the antiepileptic potential of LTG, TMS may be developed as a tool to monitor interindividual response of epilepsy patients to LTG treatment as well as to explore efficacy of other antiepileptic drugs with similar mode of action. PMID- 14529955 TI - Improved strategy for presumptive identification of methanogens using 16S riboprinting. AB - The predicted 16S riboprint patterns of 10 restriction endonucleases for 26 diverse methanogens were compared to actual patterns produced on agarose gels. The observed patterns corroborated the expected riboprints. Our analyses confirmed that the endonuclease HaeIII gave the best results generating 15 different riboprint sets. Six of these 15 riboprints represented more than one strain. Of these, three riboprint sets were further differentiated: Methanomicrobium mobile, Methanolacinia paynteri, and Methanoplanus petrolearius were differentiated from each other by the endonuclease AluI; Methanofollis liminatans, Methanospirillum hungatei, and Methanoculleus bourgensis were differentiated from each other by HpaII; and the combination of FokI and MluNI was used to differentiate Methanobrevibacter sp. ZA-10, and Methanobrevibacter arboriphilicus strains DH-1, AZ, and DC from each other. We could not differentiate the following pairs of strains from each other: Methanosarcina mazeii S-6 and C16, Methanobacterium bryantii MoH and MoH-G, Methanobacterium thermoautotrophicum GC-1 and DeltaH, and Methanobrevibacter arborophillicus DC and A2. This riboprint strategy provided a simple and rapid method to presumptively identify 22 of the 26 diverse strains of methanogens belonging to 13 genera from a range of environments. PMID- 14529957 TI - Rapid establishment of clonal isolates of freshwater autotrophic picoplankton by single-cell and single-colony sorting. AB - We describe single-cell and single-colony sorting protocols which allowed for rapid establishment of a diverse culture collection of clonal autotrophic picoplankton (APP) isolates originating from oligotrophic and oligo-mesotrophic subalpine lakes. Overall sort recoveries, expressed as the percentage of sorted microwells exhibiting APP growth, ranged from 5% to 17% depending on the type of APP, but the growth success varied greatly (from 0% to 68%) depending on the origin of the sorted sample. We applied two direct sequencing and two denaturing gradient gel electrophoresis (DGGE) protocols to identify and characterize the genetic purity of 21 of our picocyanobacteria cultures, namely, direct sequencing of the 16S rRNA gene and cpcBA-IGS region, and DGGE analyses involving a 194-bp fragment of the internal transcribed spacer (ITS) and a ca. 500-bp fragment of the phycocyanin (PC) operon (cpcBA-IGS, novel protocol described herein). Of those 21 picocyanobacteria cultures obtained by single-cell/single-colony sorting and subsequently characterized genetically/screened for genetic purity, only one culture was composed of multiple picocyanobacterial strains. PMID- 14529956 TI - Quantitative detection for low levels of Helicobacter pylori infection in experimentally infected mice by real-time PCR. AB - Accurate diagnosis of Helicobacter pylori infection is important in both clinical practice and clinical research. Molecular methods are highly specific and sensitive, and various PCR-based tests have been developed to detect H. pylori in gastric biopsy specimens. We optimized a sensitive and specific quantitative SYBR Green I real-time PCR assay for detection of H. pylori based on amplification of the fragment of a 26-kDa Helicobacter species-specific antigen gene that allows for detection of 5 bacterial cells per PCR sample. Under the assay conditions, SYBR Green I real-time PCR is highly reproducible with a precise log-linear relation in the range of six orders of magnitude of bacterial DNA concentrations. For accurate comparison of H. pylori infection in different tissue samples, the amount of total host DNA in each sample is normalized by TaqMan real-time PCR of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) pseudogenes. The developed method was validated in prophilactically immunized and experimentally infected mice and revealed a level of H. pylori gastric colonisation that was below the limit of detection for a rapid urease test. This new method established for a quantitative analysis of H. pylori in the host's stomach may be useful in experimental studies evaluating new anti-H. pylori drugs and vaccines. PMID- 14529958 TI - Comparison of signature lipid methods to determine microbial community structure in compost. AB - The microbial community structure changes substantially during the composting process and simple methods to follow these changes can potentially be used to estimate compost maturity. In this study, two such methods, the microbial identification (MIDI) method and the ester-linked (EL) procedure to determine the composition of long-chain fatty acids, were applied to compost samples of different age. The ability of the two methods to describe the microbial succession was evaluated by comparison with phospholipid fatty acid (PLFA) analysis on the same samples.Samples were taken from a 200-l laboratory compost reactor, treating source-separated organic household waste. During the initial stages of the process, the total concentration of fatty acids in compost samples treated with the EL and MIDI methods was many times higher than with the PLFA method. This was probably due to the presence of fatty acids from the organic material in the original waste. However, this substantial difference between PLFA and the other two methods was not found later in composting. Although the PLFA method gave the most detailed information about the growth and overall succession of the microbial community, the much simpler MIDI and EL methods also successfully described the shift from the initially dominating straight chain fatty acids to iso- and anteiso branched, 10 Me branched and cyclopropane fatty acids in the later stages of the process. Thus, the MIDI and EL extraction methods appear to be suitable for analysis of microbial FAME profiles in compost, particularly in the later stages of the process. PMID- 14529960 TI - Application of Amido black staining to enumerating bacteria grown on membrane filters. AB - A method to detect and enumerate bacterial colonies grown on membrane filters (MF) was described. The colonies were stained with an ethanolic solution of 0.1% Amido black 10B. The procedure yielded the rapid detection of colonies as compared to a conventional plate counting method. PMID- 14529959 TI - Comparison of different PCR tests for detecting Shiga toxin-producing Escherichia coli O157 and development of an ELISA-PCR assay for specific identification of the bacteria. AB - In an attempt to develop a standard for ELISA-PCR detection of Shiga toxin producing Escherichia coli (STEC) O157, six published PCR tests were tested in a comparative study on a panel of 277 bacterial strains isolated from foods, animals and humans. These tests were based on the detection of the genes rfbE [J. Clin. Microbiol. 36 (1998) 1801] and rfbB [Appl. Environ. Microbiol. 65 (1999) 2954], the 3' end of the eae gene [Epidemiol. Infect. 112 (1994) 449], the region immediately flanking the 5' end of the eae gene [Int. J. Food. Microbiol. 32 (1996) 103], the flicH7 gene [J. Clin. Microbiol. 35 (1997) 656], or a part of the recently described 2634-bp Small Inserted Locus (SILO(157) locus) of STEC O157 [J. Appl. Microbiol. 93 (2002) 250]. Unlike the other PCR assays, those amplifying the rfb sequences were unable to distinguish toxigenic from nontoxigenic O157. These assays were relatively specific to STEC O157, giving essentially a cross reaction with clonally related E. coli O55 and to a lesser extent with E. coli O145, O125, O126. They also detected the Shiga toxin (stx) negative derivatives of STEC O157. Based on these results, an ELISA-PCR assay consisting of the solution hybridization of amplicons with two probes that ensured the specificity of the amplification was developed. The ELISA-PCR assay, which used an internal control (IC) of inhibition, was able to detect 1 to 10 copies of STEC O157 in the PCR tube. Adaptation of PCR into ELISA-PCR assay format facilitates specific and sensitive detection of PCR amplification products and constitutes a method of choice for screening STEC O157. PMID- 14529961 TI - Computational methods and evaluation of RNA stabilization reagents for genome wide expression studies. AB - Gene expression studies require high quality messenger RNA (mRNA) in addition to other factors such as efficient primers and labeling reagents. To prevent RNA degradation and to improve the quality of gene array expression data, several commercial reagents have become available. We examined a conventional hot-phenol lysis method and RNA stabilization reagents, and generated comparative gene expression profiles from Escherichia coli cells grown on minimal medium. Our data indicate that certain RNA stabilization reagents induce stress responses and proper caution must be exercised during their use. We observed that the laboratory reagent (phenol/EtOH, 5:95, v/v) worked efficiently in isolating high quality mRNA and reproducibility was such that reliable gene expression profiles were generated. To assist in the analysis of gene expression data, we wrote a number of macros that use the most recent gene annotation and process data in accordance with gene function. Scripts were also written to examine the occurrence of artifacts, based on GC content, length of the individual open reading frame (ORF), its distribution on plus and minus DNA strands, and the distance from the replication origin. PMID- 14529962 TI - Rapid screening method for quantitation of bacterial cell lipids from whole cells. AB - Although specific lipids in bacteria can be quantitated, there is still a need to quantitate the total lipid content of a bacterial sample. The sulfo-phospho vanillin reaction for quantitation of bacterial lipids has significant advantages over traditional methods for screening of engineered mutant strains. In this report we show that this methodology can be used directly on whole cell or homogenized biological material, without any extraction step. The cell components, and most of the reagents used for cell extraction, that were tested did not interfere with the reaction. The screening is based on the observation of physiologic variations using ratios of relative amounts: lipid/DNA and lipid/protein. Our results show that significant differences in those ratios can be detected when there is a modification of the phospholipid content of the cell. The sample manipulation required is minimal and could be automated. Used as a primary screening and/or characterization of engineered mutant strain, the test may lead to further investigation of the nature and distribution of lipids in the cell. PMID- 14529963 TI - RT-PCR of long prokaryotic operon transcripts without DNase treatment. AB - RT-PCR is a powerful technique used in the amplification and detection of rare mRNAs. However, one of the most serious drawbacks of this method is the amplification of false-positive products due to DNA contamination in the RNA samples. This pitfall is particularly hard to overcome when RNA from prokaryotic origin is used. We present here a modification of the EXACT RT-PCR method that was successfully employed in the amplification of the low abundant full-length polycistronic pst operon mRNA of Escherichia coli. No DNase treatment of the RNA template is required, but unlike the original EXACT RT-PCR, a hybrid primer that is not composed of oligo(dT) was used. A nonhomologous sequence was incorporated at the reverse transcription step into the 5' end of the first-strand cDNA by means of the hybrid primer. For the PCR, a gene-specific primer and a second primer identical to the nonhomologous portion of the hybrid primer were used. To avoid amplification of genomic DNA, the hybrid-primer molecules that were not incorporated into the first-strand cDNA were removed by RNase H treatment followed by ultrafiltration. PMID- 14529964 TI - An SNP-based PCR assay to differentiate between Listeria monocytogenes lineages derived from phylogenetic analysis of the sigB gene. AB - The alternative sigma factor sigB gene is involved in the stress response regulation of Listeria monocytogenes, and contributes towards growth and survival in adverse conditions. This gene was examined to determine if it could be a useful indicator of lineage differentiation, similar to the established method based on ribotyping. The sigB sequence was resolved in four local L. monocytogenes strains and the phylogenetic relationship among these, and a further 21 sigB gene sequences from strains of different serotype and lineage including two Listeria innocua strains, obtained from the GenBank database were determined. The sigB nucleotide sequences of these 25 Listeria strains were then examined for single nucleotide polymorphic (SNP) sites that could differentiate between the three lineages. Based on nucleotide sequences L. monocytogenes lineage I/serotype 1/2b and 4b clustered together, lineage II/serotype 1/2a and 1/2c strains clustered together, lineage III/serotypes 4a and 4c strains clustered together and L. innocua strains clustered together as an outgroup. SNPs differentiating the three lineages were identified. Individual allele-specific PCR reactions based on these polymorphisms were successful in grouping known and a further 37 local L. monocytogenes isolates into the three lineages. PMID- 14529965 TI - Acquisition of flocculation phenotype by Kluyveromyces marxianus when overexpressing GAP1 gene encoding an isoform of glyceraldehyde-3-phosphate dehydrogenase. AB - The use of flocculating yeast strains has been considered as a convenient approach to obtain high cell densities in bioreactors with increasing productivity in continuous operations. In Kluyveromyces marxianus ATTC 10022, the GAP1 gene encodes an isoform of glyceraldehyde-3-phosphate dehydrogenase-p37-that is accumulated in the cell wall and is involved in flocculation. To test the use of p37 as a tool for engineering Kluyveromyces cells to display a flocculation phenotype, K. marxianus CCT 3172 was transformed with an expression vector containing GAP1. This vector is based on the pY37 previously described, harbouring a S11 Kluyveromyces origin of replication, and the expression of GAP1 is under the control of GAL1. Kluyveromyces cells overexpressing GAP1 acquired a flocculent phenotype together with the accumulation of p37 in the cell wall. The results support the use of GAP1 gene as a molecular tool for inducing flocculation. PMID- 14529966 TI - A kinetic microplate method for quantifying the antibacterial properties of biological fluids. AB - The microplate assay for measuring antibacterial activity was adapted by incorporating a known concentration range of gentamicin as an internal standard. Staphylococcus aureus subsp. aureus strain ATCC 6538P, Escherichia coli ATCC 25922, and Lactobacillus spp. were used as target organisms, although other indicator organisms and antibiotics can be examined. Assay range and sensitivity were dependent on the species and density of indicator organism, and conditions (e.g., type, concentration, and pH of growth medium). Plotting the area under the growth curve (AUGC) versus gentamicin concentration (log transformed) yielded a linear curve that was used to quantify in units of gentamicin the antibacterial activity of a secretory fluid (SCF; pancreatic juice) and for comparisons of samples collected at different times, analysed on different days, and from different studies. This adaptation of the microtiter broth method will be useful for investigating man-made compounds, and the antibacterial activity of secretory fluids and the influences of age, diet, and health status. PMID- 14529967 TI - Multiplex PCR screening of soil isolates for novel Bacillus-related lineages. AB - A16S rDNA multiplex PCR-based high-throughput protocol is presented to screen bacterial isolates in large amounts for the appearance of novel lineages of bacteria, especially hitherto unknown Bacillus relatives. The 16S rDNAs of 4224 isolates from a comprehensive cultivation campaign were screened for similarity to predominant uncultured soil bacteria. Soil suspensions were plated in serial dilutions on various media. After 2, 4 and 6 weeks, colonies were collected with toothpicks and transferred to microtiterplates for cell lysis and storage plates for subculture. Cell lysis was a simple freeze-heating cycle in distilled water. The multiplex PCR was adapted to operate sufficiently for Gram positives under these conditions. Approximately 10.6% of all picked colonies reacted with a primer targeting a Bacillus fraction containing novel Bacillus benzoevorans relatives previously detected as predominant soil bacteria by culture-independent studies. From these 446 colonies detected by multiplex PCR, 363 (81.4%) could be successfully used for continued subculture and 16S rDNA sequencing. All identification was done by 16S rDNA sequencing. This revealed that more than 60% of them represented a variety of candidates for potentially new species. Twelve colonies were identified as almost identical matches to 16S rDNA sequences of hitherto uncultured but apparently predominant soil bacteria cloned from directly extracted soil DNA. Also, novel lineages of unpredicted phylogenetic diversity like novel Paenibacillus, Sporosarcina and even Xanthomonads were represented. PMID- 14529968 TI - An efficient strategy for screening large cloned libraries of amplified 16S rDNA sequences from complex environmental communities. AB - We propose a strategy for the efficient screening of large libraries of amplified 16S rRNA genes from complex environmental samples. It consists of processing sets of multiple clones simultaneously. This strategy saves up to 90% of the costs and labor spent in the process of screening a 16S rDNA library. PMID- 14529969 TI - Evaluation of a novel method based on PCR Restriction Fragment Length Polymorphism Analysis of the tuf gene for the identification of Staphylococcus species. AB - A novel method, based on PCR Restriction Fragment Length Polymorphism Analysis (PRA) of a part of the tuf gene (370 bp), was designed for the identification of 11 staphylococcal species, including the most common staphylococcal pathogens. A total of 258 clinical isolates were validated by this assay, and the results were in concordance with those obtained by the reference method of Kloos and Schleifer. PMID- 14529970 TI - Consecutive analysis of bacterial PCR samples on a single electronic microarray. AB - For routine mass screening, the use of microarrays is hampered because one chip can only analyze one sample. 16S rRNA gene PCR products of several bacterial strains or mixtures thereof were consecutively loaded on a single electronic microarray and successfully analyzed using probes specific for the bacterial strains. PMID- 14529971 TI - A 6 x 6 drop plate method for simultaneous colony counting and MPN enumeration of Campylobacter jejuni, Listeria monocytogenes, and Escherichia coli. AB - A protocol was developed using 96-well plates and multichannel pipettes for serial dilutions, followed by drop plating on agar in a 6 x 6 format. This protocol permits simultaneous plating of six dilutions which greatly decreases the number of plates utilized thereby saving incubator space for organisms such as Campylobacter which require unique environmental conditions. PMID- 14529972 TI - Efficient transformation of the osmotolerant yeast Zygosaccharomyces rouxii by electroporation. AB - A rapid and simple electroporation method to transform osmotolerant yeast Zygosaccharomyces rouxii has been developed and conditions for efficient transformation of mutants derived from different Z. rouxii wild-type strains optimized. PMID- 14529973 TI - Challenges in reversing esophageal discontinuity operations. AB - BACKGROUND: After catastrophic esophageal or gastric disruption results in esophageal discontinuity, operations to restore swallowing are surgical challenges. METHODS: A retrospective review and assessment of functional results was performed in 40 patients (average age 59.9 years) who had 42 operations to reverse esophageal discontinuity between 1973 and 2002. RESULTS: Esophageal discontinuity resulted from gastric necrosis after esophagectomy and esophagogastrostomy (n = 10) or hiatal hernia repair (n = 4), esophageal perforation complicating dilatation (n = 5), failed colonic or jejunal interpositions (n = 5), caustic ingestion (n = 4), Boerhaave syndrome (n = 4), esophagogastric anastomotic leak (n = 3), and other causes (n = 6). Eighteen patients (43.9%) required prolonged mechanical ventilation. Thirty-one (75.6%) had an end cervical esophagostomy; 6, an anterior thoracic esophagostomy; 2, lateral esophagostomy and in situ native esophagus stapled and divided distally; and 1 each, a stapled, divided esophagus without esophagostomy and a stapled undivided esophagus without esophagostomy. Twenty-six patients (63.4%) had undergone partial or total gastrectomy. Ten (24.4%) had vocal cord paralysis. Operations reestablishing continuity included colonic interposition in 23 (56.1%), substernal gastric interposition in 7 (17.1%), esophagectomy and cervical anastomosis in 6, esophageal reanastomosis in 3, staged jejunal interposition in 1, and Roux-en-Y esophagojejunostomy in 1. There were no hospital deaths. Twenty-eight patients (68.3%) had postoperative complications. Length of stay averaged 20.6 days. Follow-up for 40 patients averaged 54.5 months. Functional results (39 patients) were excellent in 12 (30.8%), good in 15 (38.5%), fair in 10 (25.6%), and poor in 2 (5.1%). CONCLUSIONS: Successful reversal of esophageal discontinuity requires individualized assessment and ingenuity. Despite appreciable morbidity, the ultimate result is generally gratifying. PMID- 14529974 TI - Immunohistochemistry analysis of micrometastasis in pretreatment lymph nodes from patients with esophageal cancer. AB - BACKGROUND: With recent advances in neoadjuvant therapy in esophageal cancer, pretreatment lymph node staging has become increasingly important in stratifying patients to appropriate treatment regimens and for prognostication. Immunohistochemical analysis (IHC) using epithelial markers has been shown to identify micrometastases in histologically negative lymph nodes. We performed this study to evaluate if IHC analysis in thoracoscopic/laparoscopic (Ts/Ls) pretreatment staging lymph nodes can reveal additional diagnostic information to routine histopathology. METHODS: Specimens of 106 patients with esophageal cancer who had pretreatment Ts/Ls staging were retrospectively studied. Lymph node biopsies were obtained for IHC staining using cytokeratin (CK) of AE1/AE3. IHC staining for p53, an apoptosis protein associated with poor prognosis in esophageal cancer, was also performed. RESULTS: 331 Ts/Ls staging lymph node biopsies were collected from 106 patients. A total of 15.4% (51/331) of the lymph nodes or 34.9% (37/106) of patients were found to have metastatic deposits by routine histology. All the histologically positive lymph nodes were CK positive. Among the remaining 280 histologically negative lymph nodes, 11(3.9%) were found to have micrometastasis by CK staining. Three patients (4.3%, 3/69) were upstaged from N0 to N1. They died of early recurrences after treatment. A total of 67.6% (25/37) of the patients with histologically positive lymph node were p53 positive. No histologically negative lymph node was found to be p53 positive in this series. CONCLUSIONS: Immunohistochemical analysis for CK can detect micrometastatic involvement of lymph nodes that are missed on routine pathologic examination, and, therefore, can improve lymph node staging. Its clinical significance in esophageal cancer warrants further study. PMID- 14529976 TI - Video-assisted thoracic surgery lobectomy for stage I lung cancer. AB - BACKGROUND: The technique, safety, and oncologic efficacy of video-assisted thoracic surgery (VATS) lobectomy are controversial. Issues include operative time, lymph node yield, conversion to thoracotomy, resource utilization, recurrence, complications, and survival. METHODS: From January 1995 to December 2001, 179 patients underwent VATS lobectomy for preoperative stage I lung cancer (T1N0, 118 patients; T2N0, 61 patients). Mean age was 64.34 years (range, 38 to 87); 91 were female and 88 were male. Contraindications to VATS lobectomy included any suggestion of hilar, endobronchial, or central lesions. Video assisted thoracic surgery lobectomy was performed using three ports, partial anatomic hilar dissection, and mediastinal node dissection. RESULTS: Distribution of lobectomies was as follows: left upper lobe, 50 patients; left lower lobe, 27 patients; right upper lobe, 33 patients; right upper and right middle lobe, 29 patients; right middle lobe, 9 patients; right lower lobe, 30 patients; right middle lobe and right lower lobe, 1 patient. Mean operative time was 75 +/- 6 minutes. Mean lymph node yield was 11 +/- 5 nodes. Pathologic upstaging was noted in 14 of the 179 patients (7.8%). Mean hospitalization was 4.1 days (range, 2 days to 4 months). There were no conversions to thoracotomy and there was 1 death (1 of 179, 0.05%). Complications included air leak in 24 of 179 (13.4%), subcutaneous emphysema in 4 of 179 (2.2%), pneumonia in 10 of 179 (5.6%), wound infection in 5 of 179 (2.8%), respiratory failure in 3 of 179 (1.7%), pulmonary embolism in 2 of 179 (1.1%), and myocardial infarction in 1 of 179 (0.5%). At a mean follow-up of 37 months, local recurrence rate was 0.013 per person per year. Actuarial recurrence-free survival was 88% and 85% at 36 and 60 months respectively. CONCLUSIONS: For carefully selected patients VATS lobectomy for early stage lung cancer is a safe and effective strategy. Long-term follow-up is required to fully evaluate recurrence and survival. PMID- 14529975 TI - The risk of second primary tumors after resection of stage I nonsmall cell lung cancer. AB - BACKGROUND: The incidence of second primary lung cancers (SPLC) after resection of nonsmall cell lung cancer (NSCLC) is estimated to be 1% to 4% per patient year. The overall effect of SPLC on survival after resection of stage I NSCLC is unknown. Here we report the incidence, management, and outcome of SPLC in a large prospective cohort of patients who underwent careful follow-up. METHODS: National Cancer Institute Intergroup Trial NCI #I91-0001 examined the effectiveness of isotretinoin A for chemoprevention of second primary tumors, the primary endpoint in that trial. Prospective data from patients randomly assigned to the placebo arm were analyzed. RESULTS: Five hundred sixty-nine patients underwent complete resection of pathologic stage I NSCLC. The median follow-up was 5.9 years. Second primary tumors developed in 88 (15%) patients. Of these, 49 (56%) were SPLC (incidence = 1.99/100 patient-years), with a median interval from initial surgery of 4.2 years. Second primary lung cancer never developed in patients who had never smoked (n = 44, p = 0.046; never versus ever smokers). Current smokers had a higher incidence of SPLC than former smokers (hazard ratio = 1.91, p = 0.03). Age, sex, stage, histology, tumor location and initial surgery had no effect on SPLC development. Despite semiannual follow-up with chest radiographs, 12 (24%) patients had metastatic disease at the time of diagnosis of SPLC. Surgical resection was performed in 31 (63%) SPLC patients. Median survival was 4.1 years in those who underwent surgery and 1.4 years in those who did not (p = 0.003). Overall SPLC-related mortality in the original cohort was 3.7%. CONCLUSIONS: Patients who undergo surgery for SPLC can achieve prolonged survival. Despite close follow-up however many patients with SPLC present with advanced disease. That indicates a need for continued lifelong postoperative surveillance. PMID- 14529977 TI - A clinicopathological study of resected subcentimeter lung cancers: a favorable prognosis for ground glass opacity lesions. AB - BACKGROUND: Owing to the advent of refined chest computed tomography (CT) images with higher resolution and CT screening programs, more faint and smaller lung cancers are being discovered. These include small-sized lung cancers such as those with a subcentimeter diameter, which had never been picked up on the routine chest roentgenogram films. However their clinicopathological characteristics with special reference to the proper surgical mode are not fully described so far. METHODS: During a 10-year period from 1991 through 2000 a total of 1,769 lung tumors were resected at the National Cancer Center Hospital, Tokyo. According to the pathology files of these patients, 51 patients had a primary tumor with the diameter of 1 cm or less. Three tumors arising in the bronchial lumina of hilum with a squamous cell carcinoma histology were excluded and the remaining 48 tumors of peripheral origin were studied. The clinicopathological features were analyzed according to three types of appearance on high-resolution CT: non-solid ground glass opacity (GGO) type (n = 19); part-solid GGO type (n = 9); and solid type (n = 20). Non-solid GGO is made up of homogeneous moderate increased density on CT, which cannot obscure the bronchovascular structure, whereas partly solid GGO contains a mere solid part but did not exceed 50% of the whole area (n = 9). All other lesions were considered solid type. RESULTS: For the three types of lesions, the distribution of age and sex was similar with the average age of 61 years and an almost even distribution of male/female patients. Although 6 patients had symptoms, the symptoms were not associated with the nodule itself. Twenty-six patients (54%) were screen-detected (16 chest roentgenogram films and 10 CT scans) and the others were detected by incidentally taken chest roentgenogram film or CT for other reasons than nodules detected. Two squamous carcinomas were positive for sputum cytology. Preoperative cytologic/histologic diagnosis was given in 14 patients (29%). The histologic type of GGO lesion was bronchioloalveolar carcinoma in all 28 cases. In solid lesions, besides 16 adenocarcinomas 2 cases of squamous cell carcinoma, 1 case each of small cell carcinoma and carcinoid tumor was seen. Lymph node involvement was seen only in 3 patients with solid lesions (N1 in 2 patients, N2 in 1 patient). As for operative mode, the limited resection was performed for 15 GGO lesions (54%) and 4 solid lesions (20%). Tumor recurrence was seen in 2 patients with solid lesions-1 in bone and the other in locoregional lymph node, and the former died of disease. CONCLUSIONS: Among subcentimeter lung cancers, GGO lesions (both non-solid and part-solid) constitute true early lung cancers. Since they have minimal or no invasive growth, limited resection for cure is justified. Conversely the solid lesion had significant invasive features such as lymph node metastasis. Lobectomy should remain as the standard mode of surgery despite such small size. PMID- 14529978 TI - Pneumonectomy for stage I (T1N0 and T2N0) nonsmall cell lung cancer has potent, adverse impact on survival. AB - BACKGROUND: Surgically treated, stage I (T1N0 and T2N0) nonsmall cell lung cancer has a relatively favorable prognosis. Our aim was to determine whether performing a pneumonectomy in this group of patients has an impact on survival. METHODS: Four hundred eighty-five patients with stage I nonsmall cell lung cancer undergoing lung resection between 1991 and 2000 were studied. Three hundred seventy-four patients underwent a smaller resection than a pneumonectomy and 111 had a pneumonectomy. RESULTS: Patients undergoing less extensive resections were older (mean age, 65 vs 63 years) (p = 0.01); these patients were also more likely to have a history of chronic obstructive airway disease (9% vs 2%) (p = 0.01) or asthma (10% vs 3%) (p = 0.04), nonsquamous cell type (56% vs 27%) (p < 0.0001), and T1 tumor stage (66% vs 17%) (p = 0.002) than patients having a pneumonectomy. Operative mortality was 2.4% versus 8% (p = 0.01). Overall 1-, 3-, and 5-year Kaplan-Meier survival rates (95% confidence interval [CI]) after less extensive resections were 85% (CI, 82% to 90%), 63% (CI, 56% to 69%), and 50% (CI, 42% to 57%), respectively, and after pneumonectomy the survival rates were 66% (CI, 53% to 73%), 47% (CI, 35% to 57%), and 44% (CI, 32% to 55%), respectively (p = 0.0006). When the Cox proportional hazards model was applied to all study patients (n = 485), pneumonectomy (p = 0.001), T2 stage (p = 0.006), older age (p = 0.03), and male gender (p = 0.03) were independent adverse predictors of survival. When the analysis was limited to the patients having T1N0 disease (n = 145), pneumonectomy (p = 0.0008), older age (p = 0.05), and nonsquamous cell type (p = 0.02) were independent adverse determinants of survival. When only the patients with T2N0 disease were analyzed (n = 340), male gender (p = 0.0005) and pneumonectomy (p = 0.01) were independent negative predictors of survival. CONCLUSIONS: In this study, the patients who underwent pneumonectomy for stage T1N0 or T2N0 nonsmall cell lung cancer had a significantly poorer survival than those patients who underwent smaller lung resections. PMID- 14529979 TI - Intraoperative solumedrol helps prevent postpneumonectomy pulmonary edema. AB - BACKGROUND: Postpneumonectomy pulmonary edema and pneumonia are life threatening and seemingly unavoidable complications after pneumonectomy. We theorized that an intraoperative dose of intravenous steroids (as a prophylactic measure to reduce pulmonary injury to the remaining lung) just before pulmonary artery ligation might decrease this problem. METHODS: Seventy-two patients (52 men) who had pneumonectomy during two time periods were studied prospectively. Thirty-five patients received 250 mg of methylprednisolone sodium succinate (Solumedrol; Upjohn, Kalamazoo, MI) just before pulmonary artery ligation (S group) and 37 did not (non-S group). Groups were matched for known or suspected preoperative, intraoperative, and postoperative risk factors for postpneumonectomy pulmonary edema. RESULTS: The incidence of postpneumonectomy pulmonary edema or adult respiratory distress syndrome was less in the S group (0 of 35, 0% versus 5 of 37, 13.5%, p = 0.049), the overall major complication rate was less in the S group (7 of 35, 20% versus 16 of 37, 43%, p = 0.04), and the length of hospital stay was shorter in the S group (6.1 days versus 11.9 days, p = 0.02). In addition, there were no bronchopleural fistulas in the S group compared with two (both right-sided) in the non-S group. CONCLUSIONS: The intraoperative intravenous administration of 250 mg of methylprednisolone sodium succinate just before pulmonary artery ligation during pneumonectomy may reduce the incidence of postpneumonectomy pulmonary edema and adult respiratory distress syndrome as well as decrease other major complications and shorten the hospital stay. It does not seem to increase the incidence of bronchopleural fistula. Further randomized trials are needed. PMID- 14529980 TI - Pharmacokinetics after pulmonary artery perfusion with gemcitabine. AB - BACKGROUND: Isolated lung perfusion (ILuP) proved to be superior for the treatment of lung metastases compared with intravenous (i.v.) injection. However its invasive character limits repetitive treatment. Blood flow occlusion (BFO) as a regional therapy with gemcitabine (GCB) was evaluated in a rat model. Lung levels of GCB were examined with different exposure times and flow rates and compared with ILuP and i.v.. Cell kill was studied in vitro. METHODS: In vitro survival of CC531 adenocarcinoma cells was determined after 10, 20, and 40 minutes of exposure to GCB. In vivo 48 Wag/Rij rats underwent BFO with GCB at a rate of 0.2 mL/min and 0.5 mL/min during 10, 20, 30, and 40 minutes. Statistical analysis was performed using Student's t test. RESULTS: In vitro, the dose of GCB resulting in 50% growth inhibition was 9.1 microg/mL, 7.2 microg/mL, and 2.2 microg/mL after 10, 20, and 40 minutes exposure respectively. In vivo, no significant difference in lung levels of GCB was observed between a flow rate of 0.2 mL/min compared with 0.5 mL/min at any exposure time point (p < 0.05). Lung tissue was saturated after 20 minutes. Blood flow occlusion resulted in a lower plasma levels and higher lung levels of GCB compared with i.v. injection of the maximal tolerated dose of 40 mg. CONCLUSIONS: Growth inhibition of CC531 cells in vitro increased with exposure time while lung tissue was saturated after 20 minutes of BFO. No difference in GCB lung levels were seen after BFO compared with ILuP. Systemic exposure after i.v. injection was higher compared with BFO but did not result in higher lung levels. PMID- 14529981 TI - Thymic carcinoma: involvement of great vessels indicates poor prognosis. AB - BACKGROUND: Thymic carcinoma is a rare, indolent, and invasive cancer. This study investigated the treatment results of thymic carcinoma and clinical prognostic factors. METHODS: From June 1988 to January 2002, 38 patients were enrolled in this study with the diagnosis of thymic carcinoma in the Cheng-Kung University Hospital based on Rosai's and Muller-Hermelink's classification. Clinical and pathologic data were retrospectively reviewed. Survival analysis was performed using the Kaplan-Meier, log rank, and Wilcoxon tests. Statistical significance was defined as p < 0.05. RESULTS: Pathology revealed 14 poorly differentiated, 6 moderately differentiated, and 8 well-differentiated squamous cell carcinomas; 8 lymphoepithelioma-like carcinomas; and 2 other carcinomas. Pathologic staging using the Masaoka system included 6 stage II, 23 stage III, and 9 stage IV patients. Six biopsies, five debulkings, and 27 complete resections were performed. All patients were followed from 15 months to 10 years 9 months, with an average of 53.8 months. Median survival time was 81 months, and median recurrence time was 52 months. Eighteen patients are still alive, and 7 are alive with disease. Well-differentiated squamous cell carcinoma had better prognosis than other carcinomas (p = 0.022). Complete resection significantly increased survival rate (p < 0.001). Tumor invasion of the superior vena cava, pulmonary vessels, or aorta were significant predictors for poor prognosis (p = 0.016, 0.002, and 0.002, respectively). CONCLUSIONS: Only patients with thymic carcinoma who underwent complete resection had long-term survival. Prognosis of thymic carcinoma seemed mainly dependent on tumor invasion of the great vessels. PMID- 14529982 TI - Comparison of the immediate postoperative outcome of using the conventional two drains versus a single drain after lobectomy. AB - BACKGROUND: We compared the immediate postoperative outcome and cost effectiveness of using a single chest drain in the midposition to the conventional apical and basal drains after lobectomy. METHODS: Of the 120 consecutive patients who underwent thoracotomy and lobectomy for lung cancer at our center between January 2001 and December 2002, 60 had the conventional 28 French apical and basal drains (group A), whereas the remaining 60 had a single 28 French chest drain placed in the midposition before closure (group B). The assessed outcomes included length of stay, amount and duration of drainage, subcutaneous emphysema, postremoval hemothorax and pneumothorax, drain reinsertion, patient controlled analgesia duration, maximum pain scores, and analgesic usage. RESULTS: Both groups matched in terms of age (group A vs group B mean, 65 years old vs 66 years old, respectively; p = not significant [NS]) and gender (M:F, 4:1 for group A vs 4:1 for group B). There was no significant difference in the length of stay (mean, 7.7 days for group A vs 7.8 days for group B; p = NS), amount of drainage (mean, 667 mL for group A vs 804 mL for group B; p = NS), duration of drainage (mean, 4 days for group A vs 4.3 days for group B; p = NS), duration of patient controlled analgesia (mean, 3.7 days for group A vs 4.2 days for group B; p = NS) and analgesic combinations used (nonsteroidal antiinflammatory drugs +/- oral opioids +/- paracetamol) between the two groups. There were no clinically significant postdrain removals of hemothorax or pneumothorax in either group. Group A patients had a significantly higher maximum pain score compared with group B patients (mean, 1.4 vs 1.02, respectively; p = 0.02). Cost savings per patient in group B was more than or equal to 55 US dollars, which added up to a total cost savings of approximately more than or equal to 3,300 US dollars. CONCLUSIONS: A single chest drain in the midposition is just as effective, significantly less painful, and much more cost effective than the conventional use of two drains after lobectomy. PMID- 14529983 TI - The effect of standard posterolateral versus muscle-sparing thoracotomy on multiple parameters. AB - BACKGROUND: Different alternative approaches to thoracotomy have been developed because of the considerable morbidity associated with the standard posterolateral incision. METHODS: We studied a prospective, randomized, blinded study of 60 consecutive patients to compare surgical approach time, postoperative pain (quantitated by narcotic requirements and the visual analogue scale), pulmonary function, shoulder strength, and range of motion between standard posterolateral (group I) and muscle-sparing (group II) thoracotomy techniques. RESULTS: There were no differences in postoperative surgical time, pulmonary function, shoulder range of motion, mortality, or hospitalization time. There was significantly less postoperative pain in group II. In this group, narcotic requirement was less in the first 24 hours, and visual analogue scale scores were significantly lower (p < 0.05) throughout the first postoperative week. Muscle strength had returned to preoperative levels by 1 month in both groups. Morbidity was identical in the two groups with the exception of postoperative seromas. The prevalence of seroma was 16.6% in the muscle-sparing group. CONCLUSIONS: We conclude that the muscle sparing incision may be a sensible alternative to a standard posterolateral thoracotomy. PMID- 14529984 TI - A prospective, double-blinded, randomized trial evaluating the use of preemptive analgesia of the skin before thoracotomy. AB - BACKGROUND: Thoracic surgeons spend a lot of time treating the pain of thoracotomy. METHODS: A total of 119 consecutive patients underwent elective thoracotomy. They were prospectively randomized into two groups. One group received an injection of 1% lidocaine with epinephrine in the planned skin incision just before thoracotomy, and the other group received an equal amount of saline and epinephrine. All patients had a functioning preoperative epidural; a skin incision the width of their latissimus dorsi muscle, which was cut; sparing of the serratus anterior muscle; undercutting of the sixth rib; intercostal nerve blocks before rib spreading; a similar number of chest tubes and pulmonary resections; and comparable postoperative pain management. Pain was objectified by a numeric pain score, a visual pain score, and by the Modified McGill pain questionnaire each day in the hospital, and at 3, 6, and 12 months postoperatively. RESULTS: There were 66 patients in the lidocaine group (L group) and 53 patients in the saline group (S group). Although a trend was noted toward less pain in the L group during the first 3 postoperative days (on the numeric pain scale only) the difference was not statistically significant in overall pain in the hospital or at 3, 6, and 12 months after the operation. Other pain scores and descriptors were similar throughout. CONCLUSIONS: The injection of lidocaine and epinephrine in the skin just before thoracotomy does not decrease the amount or type of pain during the hospital stay or at 3, 6, and 12 months after surgery. PMID- 14529985 TI - Tetralogy of Fallot with subarterial ventricular septal defect. AB - BACKGROUND: Tetralogy of Fallot with subarterial ventricular septal defect is frequently seen among Asians. Compared with infracristal ventricular septal defect, postoperative right ventricular outflow obstruction is more likely because of subpulmonary extension of the defect. Moreover the incidence of aortic regurgitation is a concern because of the absence of a supporting infundibulum. METHODS: Four hundred cases of classic tetralogy were reviewed, 61 of which had subarterial ventricular septal defect. RESULTS: Aortic regurgitation (of more than mild degree) was identified in 7 cases with subarterial and 7 with infracristal ventricular septal defects. The mechanism of infracristal defect was predominantly an annular dilation before surgery. In contrast 5 cases with subarterial defect had progression of aortic regurgitation after operation yielding an actuarial incidence of 29.7% at 20 years. In 2 patients the cause seemed to be fixation or plication of the aortic valve annulus by the ventricular septal patch. Compared with infracristal defect, subarterial defect was associated with increased incidence of reoperation (12.0% versus 1.9% at 10 years, p = 0.01), frequent use of transannular patch (70.5% versus 45.7%, p = 0.0004), and worse New York Heart Association (NYHA) functional class (p = 0.007). Right ventricular outflow obstruction was the reason for reoperation in 3 patients with subarterial defect and was associated with worse NYHA classification in the long-term, on multivariate analysis (p = 0.0002). CONCLUSIONS: Tetralogy with subarterial ventricular septal defect was associated with worse functional outcome. To prevent adverse outcomes, precise suturing of the distal ventricular septal patch, extensive infundibulectomy, lower threshold for transannular incision, and smaller-sized ventricular septal patch placement are warranted. PMID- 14529986 TI - Expanding the indications for pulmonary valve replacement after repair of tetralogy of fallot. AB - BACKGROUND: Insertion of a competent pulmonary valve has been advocated to reduce right ventricular volume overload associated with pulmonary regurgitation (PR) after repair of tetralogy of Fallot. However the indications, proper timing, and long-term benefits of restoring pulmonary valve function remain controversial. METHODS: Thirty-six patients (aged 15.2 +/- 9.2 years) underwent pulmonary valve implantation (31 homografts, 5 heterografts) 12.2 +/- 6.9 years after tetralogy repair. Additional surgical procedures included pulmonary artery augmentation (n = 14), closure of septal defects (n = 10), and cryoablation and endocardial resection of ventricular tachycardia (n = 2). RESULTS: All patients have had clinical improvement in their exercise capacity. Preoperative and postoperative bicycle ergometry tests in 6 patients demonstrated significant improvement in the percent of predicted peak workload (68.5% +/- 19.8% to 80.7% +/- 17.4%, p < 0.015). One midterm death occurred in a 38-year-old patient with a history of ventricular tachycardia who died suddenly 2 years after pulmonary valve insertion. Postoperative echocardiographic measurements were available in 34 patients at a mean follow-up of 5 years. There was a 30% reduction in right ventricular end-diastolic diameter indexed to body surface area after surgery (30.1 +/- 10.2 to 18.6 +/- 6.0 mm/m(2), p < 0.0001). Two patients required conduit replacements at 1 and 9 years postoperatively. CONCLUSIONS: Timely insertion of a competent pulmonary valve in children, adolescents, and young adults with significant PR after tetralogy of Fallot repair results in subjective and objective improvement in exercise capacity and is associated with reduction in right ventricle size. PMID- 14529988 TI - Liberal use of tricuspid valve detachment for transatrial ventricular septal defect closure. AB - BACKGROUND: Although temporary tricuspid valve detachment is useful for improved visualization of ventricular septal defect through right atriotomy, liberal use of this adjunct is not widely supported, mainly because of concerns about iatrogenic complications such as heart blocks and tricuspid valve dysfunction. The objective of this study was to determine whether liberal use of this adjunct can improve operative outcome. METHODS: Between January 1997 and March 2002, trans-atrial closure of isolated ventricular septal defect (conoventricular or canal type) was performed in 87 consecutive patients. Tricuspid valve detachment was used in 4 out of 44 patients (prudent-use group) and 19 out of 43 patients (liberal-use group) in the first and second half of this period, respectively (p = 0.0002). Patient demographics and use of other surgical and cardiopulmonary bypass techniques remained virtually unchanged during this period. RESULTS: In the prudent-use group, there was one operative death with prolonged bypass time and one residual defect that required reoperation; neither of these patients underwent tricuspid valve detachment. All other patients (both groups) were free from mortality and clinically significant complications, including heart block, tricuspid regurgitation, and residual defect. The liberal-use group had shorter cardiopulmonary bypass time than the prudent-use group (59 +/- 14 vs 67 +/- 22 minutes, p = 0.037). CONCLUSIONS: Tricuspid valve detachment should be used liberally for moderate- or even low-difficulty exposure of ventricular septal defect, regardless of patient background, because it is a safe and effective adjunct that can improve speed, programmability, reproducibility, and reliability. PMID- 14529989 TI - Comparative long-term results of surgery versus balloon valvuloplasty for pulmonary valve stenosis in infants and children. AB - BACKGROUND: We compared the long-term results of surgical valvotomy (S) versus balloon valvuloplasty (BV) for pulmonary valve stenosis in infants and children. METHODS: Results after surgical pulmonary valvotomy (with concomitant ASD/VSD closure) (n = 62, age 2.9 +/- 3.5 years) and balloon valvuloplasty (n = 108, age 3.6 +/- 3.9 years) were analyzed. Transvalvular mean pressure gradient decrease, freedom from reintervention for restenosis, pulmonary valve insufficiency, and tricuspid valve insufficiency were considered. RESULTS: Mean pressure gradient decreased significantly more in the surgical group (from 64.8 +/- 30.8 mm Hg to 12.8 +/- 9.8 mm Hg at a mean follow-up of 9.8 years) than after BV (decreasing from 66.2 +/- 21.4 mm Hg to 21.5 +/- 15.9 mm Hg after a mean of 5.4 years; p < 0.001). Moderate pulmonary valve insufficiency occurred in 44% after surgery, and in 11% after BV (p < 0.001). Tricuspid valve insufficiency occurred in 2% after surgery, and in 5% after BV. Restenosis occurred in 3 surgical patients (5.6%), 2 patients required reoperation, and 1 patient required a balloon valvotomy. Restenosis developed in 13 BV patients (14.1%): 6 patients were redilated and 7 patients required surgery. Surgical valvotomy led to significantly less reinterventions than balloon valvuloplasty (p < 0.04). CONCLUSIONS: Surgical relief of pulmonary valve stenosis produces lower long-term gradients and results in longer freedom from reintervention. Balloon valvuloplasty may remain, despite these results, the preferred therapy for isolated pulmonary valve stenosis, because it is less invasive, less expensive, and requires a shorter hospital stay. Surgery should remain the exclusive form of therapy in the presence of concomitant intracardiac defects, which need to be addressed. PMID- 14529990 TI - Early experience with a modified Norwood procedure using right ventricle to pulmonary artery conduit. AB - BACKGROUND: A recent modification to the Norwood procedure involves the use of a right-ventricle (RV) to pulmonary artery (PA) conduit to provide pulmonary blood flow for patients with hypoplastic left heart syndrome (HLHS). This modification is thought to provide more stable hemodynamics by avoiding the diastolic "run off" that occurs with a Blalock-Taussig shunt. METHODS: We reviewed our experience with the first 11 patients undergoing the RV-PA conduit modification of the Norwood operation and compared their outcomes with those of the preceding 22 patients who underwent a conventional Norwood procedure. RESULTS: Between July 1999 and March 2002, 33 patients with HLHS underwent the Norwood procedure at a median age of 5 days (range 1 to 31 days). Aortic atresia was present in 28 (85%). No significant difference was noted between the RV-PA (n = 11) and conventional Norwood (n = 22) groups with respect to measures of morbidity such as duration of mechanical ventilation or hospital stay. Patients who underwent the conventional Norwood procedure did have significantly lower diastolic blood pressure in the early postoperative period (38.4 +/- 4.4 mm Hg versus 49.5 +/- 4.3 mm Hg, p = 0.001). The operative and 1-year survival rates were 81% and 81%, respectively, for patients with the RV-PA modification, which was not significantly different from those of patients who underwent the conventional procedure, 81% and 73% (p = 1.00 and p = 0.36). Two patients developed a pseudoaneurysm of the RV infundibulum after placement of RV-PA conduit. Four sudden deaths occurred after hospital discharge, all occurring in the conventional Norwood group. CONCLUSIONS: The RV-PA conduit modification of the Norwood procedure results in excellent early survival. By avoiding low diastolic blood pressure this modification may provide superior perfusion to the coronary vascular bed and potentially reduce the risk of sudden unexpected death. PMID- 14529992 TI - Transverse arch hypoplasia may predispose patients to aneurysm formation after patch repair of aortic coarctation. AB - BACKGROUND: Thoracic aortic aneurysm after patch repair of aortic coarctation is a potentially lethal complication. We hypothesized that transverse arch hypoplasia in association with patch repair of aortic coarctation predisposes aneurysm formation. METHODS: A retrospective analysis was performed on all patients undergoing isolated aortic coarctation repair at the University of Virginia Health Systems between 1970 and 1995. Of 244 repairs, 38 patients underwent patch repair. These 38 patients were divided into two groups. The aneurysm group (n = 15) had aortic aneurysms develop at the repair site, which required aneurysmectomy. The nonaneurysm group (n = 23) did not have any aneurysms develop. Univariate analysis and Fisher's exact text were used to identify factors that independently predict aneurysm formation. RESULTS: Univariate analysis demonstrated aortic arch hypoplasia associated with patch repair independently predicts future aneurysm formation (p < 0.01). Patients who had an aneurysm develop also had a similar incidence of bicuspid aortic valves, mild restenosis, and late hypertension compared with patients in the nonaneurysm group. CONCLUSIONS: Aneurysm formation after patch repair of aortic coarctation develops into a subgroup of patients. Aortic arch hypoplasia associated with coarctation independently predicts future aneurysm formation. PMID- 14529993 TI - Mild to moderate mitral regurgitation in patients undergoing coronary bypass grafting: effects on operative mortality and long-term significance. AB - BACKGROUND: Patients undergoing bypass grafting (CABG) often present with mitral regurgitation (MR). While surgical strategy for patients with either trace or severe MR is well established, the need for a valve procedure with mild (2) to moderate (3+) mitral regurgitation is controversial. METHODS: We reviewed 1,939 consecutive CABG patients (1987 to 1999). A preoperative echocardiogram performed when clinically indicated graded MR from 1 to 4+. Patient characteristics, hospital mortality, and long-term survival were compared between 167 patients with grade 2 to 3+ MR and controls. A multivariate analysis identified independent predictors for long-term mortality. RESULTS: The MR patients were more often female and older; had increased comorbidities including hypertension, diabetes, and heart failure; had more extensive coronary disease and worse left ventricular (LV) function; and required urgent surgery more often. Operative mortality was 0.8% in no MR patients and 1.8% in MR patients (p not significant). Long-term survival for MR patients with poor LV function (LV grade 3 to 4) was significantly lower (53% versus 75% at 10 years, p = 0.001). Independent predictors of poor long-term survival were advanced age, LV dysfunction, heart failure, diabetes, prior cerebrovascular accident, peripheral vascular disease, and no left internal mammary artery use. CONCLUSIONS: Coronary artery bypass graft patients with mild or moderate MR have worse baseline characteristics but operative mortality with CABG alone is not significantly increased. Long-term prognosis for MR patients with poor LV function is worse compared with patients with no MR but MR was not an independent predictor of long-term mortality. To determine whether surgical correction of MR would improve results, a prospective randomized trial seems warranted. PMID- 14529994 TI - Allogeneic blood transfusion requirements after minimally invasive versus conventional aortic valve replacement: a risk-adjusted analysis. AB - BACKGROUND: Aortic valve replacement (AVR) through a partial sternotomy (mini AVR) has been suggested to significantly reduce postoperative morbidity compared with conventional AVR. This study sought to investigate whether mini-AVR patients require fewer transfusions than patients who had conventional AVR. METHODS: Of 511 patients who had AVR, 56 had mini-AVR and 455 had conventional AVR. A matched case logistic regression analysis was used to adjust for these imbalances between groups. RESULTS: No patient in the mini-AVR cohort required conversion to a conventional AVR. Cardiopulmonary bypass time was longer in the mini-AVR group compared with the conventional AVR group, with a median of 102 minutes (range, 78 to 119 minutes) versus 75 minutes (range, 61 to 96 minutes; p < 0.01) in the conventional AVR group. A total of 31 patients (55%) in the mini-AVR group and 336 patients (74%) in the conventional sternotomy group required transfusions during their hospital stay (p < 0.01). After adjusting for differences in preoperative risk factors, year of operation, and surgeon, by matching on propensity score, the differences were not statistically significant (odds ratio = 0.84, 95% confidence interval = 0.40 to 1.75, p = 0.63). CONCLUSIONS: Mini-AVR produces better wound cosmesis and less surgical trauma but requires more time to perform. Matched-case analysis failed to show a significant difference in blood transfusion requirements after mini-AVR compared with the conventional AVR approach. PMID- 14529995 TI - Mass regression in aortic stenosis after valve replacement with small size pericardial bioprosthesis. AB - BACKGROUND: The aim of the study was to determine whether left ventricular mass regression is influenced by valve size after the implantation of a Carpentier Edwards Perimount (CEP) pericardial bioprosthesis for pure aortic stenosis. METHODS: Patients receiving 19-mm, 21-mm, and 23-mm CEP aortic valves underwent echocardiography preoperatively and at least 1 year after surgery (mean, 2.3 +/- 1 years) and the echocardiograms were compared within and between groups. RESULTS: The study involved a total of 88 patients: 34 receiving 19-mm CEPs, 29 receiving 21-mm CEPs, and 25 receiving 23-mm CEPs. The mean postoperative prosthetic gradients were respectively 20.6 +/- 6.6 mm Hg, 17.9 +/- 5.8 mm Hg, and 13.2 +/- 4.1 mm Hg (p = 0.0001); the mean postoperative valve areas were respectively 1.24 +/- 0.16 cm(2), 1.45 +/- 0.2 cm(2), and 1.63 +/- 0.21 cm(2) (p = 0.0001). In comparison with the preoperative echocardiographic measurements absolute left ventricular mass significantly decreased by -54.1 +/- 48.8 g, -54.1 +/- 55.1 g, and -74.4 +/- 57.4 g respectively with no statistically significant between-group difference (analysis of variance) but ventricular septum and posterior wall thickness significantly decreased in each group (p < 0.05). CONCLUSIONS: The implantation of 19-mm, 21-mm, and 23-mm CEP aortic prostheses significantly reduces left ventricular mass without any size-related differences. PMID- 14529996 TI - Cardiac surgery in patients infected with the human immunodeficiency virus. AB - BACKGROUND: Highly active antiretroviral therapy has dramatically impacted the natural history of human immunodeficiency virus (HIV) infection and may be associated with lipodystrophy and accelerated coronary artery disease. Patients with HIV are consequently increasingly likely to present for cardiac surgery. METHODS: A retrospective review of 37 consecutive patients at two integrated centers from 1994 to 2000 was conducted. Standard database and follow-up information was supplemented with data on opportunistic infections, CD4 count, viral load, New York Heart Association status, and angina status. Risk to operating room personnel was also reviewed. RESULTS: Median age was 41 years; 34 of 37 patients were male. Operations performed were coronary artery bypass graft ([CABG] 27), aortic valve replacement ([AVR] 4), AVR/CABG (2), AVR/mitral valve repair (1), mitral valve repair (1), excision of atrial masses (1), and tricuspid valve repair (1). Complications included death in 1 of 37 (2.7%), sepsis in 2 of 37 (5.4%), deep sternal infection in 1 of 37 (2.7%), bleeding in 2 of 37 (5.4%), prolonged ventilation in 2 of 37 (5.4%), and readmission in 8 of 37 (21.6%). Actuarial freedom from a composite end point of angina, death, myocardial infarction, repeat revascularization, and congestive heart failure was 81% at 3 years with no late deaths. Preoperative and follow-up CD4 counts and viral loads were not significantly different at a mean follow-up of 28 months. No patients progressed from HIV positive status to AIDS during the study period. Six "needle stick" injuries requiring antiretroviral prophylaxis occurred in 5 caregivers without seroconversion. CONCLUSIONS: In selected patients infected with HIV, risks and outcomes of cardiac surgery are acceptable. With concomitant highly active antiretroviral therapy, intermediate HIV and cardiac status appear to be favorable. Needle stick injuries occur at a rate mandating optimal reduction of patient viral loads preoperatively. PMID- 14529997 TI - Gender influence on cognitive function after cardiac operation. AB - BACKGROUND: Women are at higher risk than men for stroke after cardiac operation. The purpose of this study was to evaluate for gender influences on the more common postoperative neurologic complication, cognitive dysfunction. METHODS: A standard battery of neuropsychological tests was administered to 117 patients (79 men and 38 women) the day before and again 4 to 6 weeks after cardiac operation. The battery assessed a broad array of cognitive domains, including attention, memory, executive function, and psychomotor processing speed. Analysis was performed only on patients with data from both testing sessions. Data were analyzed to assess for a dichotomous definition of postoperative cognitive impairment and to evaluate for factors influencing test results for specific cognitive domains. RESULTS: The frequency of one standard deviation decline on two or more cognitive tests compared with preoperative results (women, 10.7 % versus men, 9.9 %; p = 0.527), no decline, or one standard deviation improvement on each test postoperatively was no different between genders. After adjusting for age, gender, preexisting medical conditions, level of attained education, preoperative cognitive tests results, type of operation, and duration of cardiopulmonary bypass, female gender was independently associated with poorer performance postoperatively on visuospatial tasks. Other variables significantly related to postoperative cognitive function varied among the specific cognitive domains. CONCLUSIONS: These data suggest that, although the frequency of cognitive dysfunction after cardiac operation is similar for women and men, women appear more likely to suffer injury to brain areas subserving visuospatial processing. Risk factors for postoperative cognitive impairment vary depending on cognitive domain, suggesting multiple etiologies for this form of perioperative neurologic injury. PMID- 14529998 TI - Expanded use of suction and stabilization devices in cardiothoracic surgery. AB - BACKGROUND: Recent developments in beating heart instrumentation have allowed surgeons to perform multivessel off-pump coronary artery bypass surgery by overcoming the factors that impair surgical performance (limited visualization, continuous movement of the target area, and hemodynamic instability during exposure of the inferior and posterior parts of the heart). We have explored the possibility of further expanding apical and epicardial suction devices beyond their ordinary use in coronary artery surgery. METHODS: A retrospective review of our cardiac surgery database was undertaken to identify the incidences in which these devices have been used in a somewhat novel manner in the field of cardiac surgery. RESULTS: During this time period we identified 20 instances in which either apical or epicardial suction devices were used alone or together. The nature of the procedures included dividing pericardial adhesions (pericardiectomy, n = 2; dividing adhesions in redo-coronary surgery, n = 4), securing epicardial hemostasis (penetrating cardiac trauma, n = 2; securing hemostasis during reexploration after cardiac surgery, n = 10), and facilitating epicardial microwave ablation (n = 2). On all 20 occasions the intended procedures were undertaken uneventfully. Furthermore, the use of apical and epicardial devices greatly facilitated the operative procedures and avoided the use of cardiopulmonary bypass on all occasions. CONCLUSIONS: In summary, we would like to highlight the expanding role for apical and epicardial suction devices to an area beyond coronary artery surgery in which we have found their use invaluable. PMID- 14529999 TI - Do hospitals with low mortality rates in coronary artery bypass also perform well in valve replacement? AB - BACKGROUND: While hospital performance in coronary artery bypass graft (CABG) surgery is reported widely, patients may find it difficult to learn about their hospital's performance in heart valve replacement. We sought to determine if a hospital's performance in CABG is correlated to its performance in heart valve replacement. METHODS: We studied operative mortality after CABG, aortic valve replacement (AVR), and mitral valve replacement (MVR) using the 1994 to 1999 national Medicare database. After excluding any hospital that did not perform at least 50 CABGs and 20 valve replacements per year we examined the correlation between hospital mortality in CABG and hospital mortality in AVR and MVR using least-squares simple linear regression models. Operative mortality was adjusted for patient characteristics using logistic regression models. RESULTS: A total of 684 hospitals performed 817,606 isolated CABGs, 142,488 AVRs (54% with concomitant CABG), and 61,252 MVRs (45% with concomitant CABG). Hospital mortality rates with AVR ranged from 6.0% to 13.0% between hospitals in the lowest and highest, respectively, 10th percentile of CABG performance. Similarly hospital mortality rates with MVR ranged from 10.1% to 20.5% in the lowest and highest respectively, 10th percentile of CABG performance. Adjusted mortality rates for both AVR and MVR were closely correlated with isolated CABG mortality rates (correlation coefficients 0.592 and 0.538, respectively; p = 0.001 for both correlations). In stratified analyses these correlations persisted regardless of whether valve replacement was performed with or without concomitant CABG or whether valve replacement was performed in a high- or low-volume hospital. CONCLUSIONS: Hospital mortality rates with CABG are closely correlated with mortality rates with valve replacement. These findings suggest that shared processes and systems of care are important determinants of performance in cardiac surgery. PMID- 14530000 TI - Conversion in off-pump coronary artery bypass grafting: an analysis of predictors and outcomes. AB - BACKGROUND: The incidence, predictive factors, and outcomes related to conversion from off-pump coronary artery bypass (OPCAB) to on-pump coronary artery bypass grafting (ONCAB) have not been well defined. We sought to determine the incidence of conversion, predictive factors, and any associated adverse consequences. METHODS: From January 2000 through June 2002, 1,644 patients underwent nonemergent OPCAB with 61 patients requiring conversion from OPCAB to ONCAB. These groups were retrospectively compared by univariate and multivariate regression analysis. The converted group was then computer matched 1:3, to a cohort of ONCAB patients to determine differences in outcomes. RESULTS: The overall conversion rate was 3.71%. Converted patients compared with a computer matched ONCAB patients had a higher incidence of operative mortality (18.0% versus 2.7%, p < 0.001). Urgently converted patients had a higher incidence of postoperative cardiac arrest (25% versus 1.1%, p < 0.001), multisystem organ failure (10.7% versus 0.6%, p < 0.001), vascular complications (7.1% versus 1.1%, p = 0.03), and perioperative myocardial infarction (10.7% versus 1.1%, p = 0.02). Predictive factors for conversion were surgeon early in OPCAB experience (odds ratio [OR] 4.4), previous CABG (OR 2.8), and congestive heart failure (OR 2.0). The need for urgent-emergent conversion was highly predictive for operative mortality (OR 7.3) compared with elective conversion. CONCLUSIONS: Patients undergoing urgent-emergent but not elective conversion from OPCAB to ONCAB had a significantly higher risk of mortality and morbidity compared with patients whose procedure was initially ONCAB. Variables predictive of conversion included previous CABG, congestive heart failure, and surgeons early in OPCAB experience. PMID- 14530001 TI - Heparin-coated circuits and reduced systemic anticoagulation applied to 2500 consecutive first-time coronary artery bypass grafting procedures. AB - BACKGROUND: In contrast to the widespread popularity of off-pump techniques for coronary artery bypass grafting, our institution has chosen a different strategy, emphasizing improvements in the technology for extracorporeal circulation, as well as simplifying surgical and clinical management. The clinical short-term results of this approach were analyzed. METHODS: The on-pump strategy includes routine use of heparin-coated circuits combined with low systemic heparinization (activated coagulation time of more than 250 seconds), intention of total revascularization within limited ischemic times and pump times, minimal use of blood transfusions, early extubation, and rapid postoperative recovery. The data from the first 2,500 consecutive first-time coronary artery bypass grafting patients (January 1998 to February 2002) treated with this protocol were retrospectively analyzed. RESULTS: There were 487 female (median age 68 years) and 2013 male (median age 64 years) patients. A median of four (one to nine) (mean 4.5 +/- 1.2) distal anastomoses were created, and the median aortic cross clamp time and pump time were 34 and 54 minutes, respectively. At least one internal mammary artery was used in 99.7% of the patients. Blood or bank blood products were given to 118 patients (4.7%). Median extubation time was 1.5 hours. The stroke rate was 0.8%, transient neurologic deficits occurred in 0.6% of the patients, and the incidence of perioperative myocardial infarction was 1.1%. By the fifth day, 91% of the patients were ready for discharge. Seven patients (0.28%) died during their hospital stay. CONCLUSIONS: Coronary artery bypass grafting with heparin-coated cardiopulmonary bypass circuits and reduced systemic anticoagulation resulted in excellent clinical results, with minimal blood transfusions and rapid postoperative mobilization. The high number of grafted coronary arteries indicates complete revascularization in most patients, which is known to be a significant predictor of long-term event-free survival. PMID- 14530003 TI - Coronary artery bypass grafting in type II diabetic patients: a comparison between insulin-dependent and non-insulin-dependent patients at short- and mid term follow-up. AB - BACKGROUND: Diabetes is a well-established risk factor for coronary artery disease, and it is associated with an increased rate of early and late adverse events after myocardial revascularization by coronary artery bypass grafting. METHODS: A prospective follow-up study was done to evaluate the short-term and mid-term outcomes of type II diabetic patients who had coronary artery bypass grafting at our institution between 1996 and May 1999. A total of 200 patients, 100 insulin-dependent diabetic patients (group I) and 100 non-insulin-dependent diabetic patients (group II), met the inclusion criteria of the study and were included in the clinical follow-up study. RESULTS: The characteristics of the patients of the two groups were similar for baseline clinical angiographic and operative characteristics. In particular, no significant differences in cardiopulmonary bypass and aortic cross-clamping times were noted between the two groups. The number grafts per patient was similar between the two groups. There were no in-hospital deaths, but postoperative complications were different among the two series. In fact, 33% of patients in group I had at least one major complication compared with 20% in group II (p = 0.037). The cumulative number of complications was 148 in group I and 69 in group II, and the mean number of complications per patient was 4.5 and 3.5 in groups I and II, respectively. The major differences in perioperative complication rates were found in the need for prolonged (> 24 hours) ventilation, occurrence of respiratory or renal insufficiency, and mediastinitis. The mean length of stay in the intensive care unit and for total hospitalization were longer in group I than group II (4.3 +/- 2.8 days versus 2.8 +/- 2.7 days [p = 0.010] and 11.1 +/- 2.2 days versus 7.2 +/- 2.4 group II [p < 0.05], respectively). At long-term follow-up, group I patients had a significantly higher mortality rate (29% versus 10%, p < 0.001). Moreover, overall late cardiac and noncardiac complication rates were significantly higher in group I than II (37% versus 22%, p = 0.02). In the multivariate analysis including several preoperative and operative variables, treatment by insulin, advanced age (> 75 years), left ventricular dysfunction (left ventricular ejection fraction < 35%), and complex lesions at coronary angiography (American Heart Association lesion classification type C lesion) were found as independent predictors of increased mortality. CONCLUSIONS: Our data show that patients with insulin-dependent type II diabetes who had coronary artery bypass grafting have a significantly higher rate of major postoperative complications with an extremely unfavorable short- and long-term prognosis. Diabetic patients on insulin treatment should be considered high-risk candidates for coronary artery bypass grafting and require intense perioperative and long-term monitoring. Further studies will be necessary to investigate whether such conclusions may be appropriate for newer surgical strategies such as off-pump operation. PMID- 14530004 TI - Is the hospital volume-mortality relationship in coronary artery bypass surgery the same for low-risk versus high-risk patients? AB - BACKGROUND: There is evidence to support the existence of an inverse relation between mortality after coronary artery bypass graft (CABG) surgery and procedure volume. It is unclear whether all patients benefit equally from having CABG surgery performed at high-volume centers. The objective of this study was to determine whether the volume-outcome association for CABG surgery is modified by patient risk. METHODS: This retrospective cohort analysis was conducted using data from the Cardiac Surgery Reporting System database on all patients (20,078) undergoing CABG surgery in New York State who were discharged in 1996. The main outcome measure was in-hospital mortality as a function of procedure volume after adjusting for severity of disease. Logistic regression modeling was used to explore the interaction between patient risk and procedure volume. RESULTS: There is a significant interaction between procedure volume and patient risk (p = 0.01). The final model exhibits excellent discrimination (C statistic = 0.818) and goodness-of-fit (Hosmer-Lemeshow statistic = 6.02; p = 0.645). Very low (<0.5%) and low-risk (0.5%-2.0%) patients exhibit a greater reduction in CABG mortality than high (5.0%-10.0%) and very high risk (>10%) patients at high volume centers relative to low-volume centers. Among the highest risk patients (>25% risk of mortality), higher risk patients have better outcomes at higher volume centers. CONCLUSIONS: For the vast majority of patients, low-risk patients benefit significantly more than high-risk patients from undergoing CABG surgery at high-volume centers instead of at low-volume centers. Low-risk patients benefit significantly more than high-risk patients from undergoing CABG surgery at high-volume centers instead of at low-volume centers. However, before generalizing these findings to other states, this study should be repeated using other regional population-based clinical databases. PMID- 14530005 TI - Mid-term results of combined transmyocardial laser revascularization and coronary artery bypass. AB - BACKGROUND: Transmyocardial laser revascularization is increasingly used to treat intractable angina in the absence of graftable vessels; however, its role in combination with coronary artery bypass grafting remains undefined. The aim of this pilot study was to investigate the outcome of the combination therapy at mid term follow-up. METHODS: Patients (n = 20) who had elective coronary artery bypass with one or more nongraftable coronary arteries were prospectively randomized to have either coronary artery bypass grafting alone or combination coronary artery bypass grafting plus transmyocardial laser revascularization with a holmium:YAG (yttrium-aluminum-garnet) laser to nongraftable areas. All patients had an exercise tolerance test preoperatively and at 6, 18, and 36 months follow up. Stress echocardiography was performed on 17 patients at 18 months postoperatively, and regional wall motion score index was calculated in lased and nonlased nonrevascularizable myocardium of the left ventricle at rest and with dobutamine stress. RESULTS: Both groups of patients were similar in preoperative demographics and operative data. There was no perioperative death. There was no difference between the two groups in angina scoring at 6, 18, and 36 months follow-up. Exercise tolerance improved by a mean of 46.8 +/- 20.0 seconds in the coronary artery bypass grafting group versus 199.2 +/- 66.5 seconds per patient in the coronary artery bypass grafting plus transmyocardial laser revascularization group (p = 1.8 x10(-6)) at 6 months; this benefit was maintained at 18 months (157 +/- 46.3 versus 61 +/- 39.2 seconds; p = 4 x10(-4)) but was lost at 36 months (57.2. +/- 42.1 versus 68.1 +/- 46.7 seconds; p = 0.70). The mean values for wall motion score index in the lased and nonlased regions at each stage of dobutamine stress at 18 months after surgery were not statistically significant. CONCLUSIONS: The combination of coronary artery bypass and transmyocardial laser revascularization improved exercise tolerance in patients in whom complete revascularization could not be achieved by bypass grafting alone in the short term, but this benefit was lost by 36 months postoperatively. The transient improvement in exercise tolerance cannot be explained by changes in contractility in the lased areas. PMID- 14530006 TI - Continuous axial-flow left ventricular assist device (Jarvik 2000) maintains kidney and liver perfusion for up to 6 months. AB - BACKGROUND: The Jarvik 2000 axial flow left ventricular assist device (LVAD), under development for the past decade, has the potential to support patients temporarily until cardiac transplantation or as a permanent circulatory support, without the size limitations of other implantable systems. METHODS: To assess its ability to perfuse the kidneys and liver, we monitored renal and hepatic function in 10 patients who received the Jarvik 2000 LVAD as a bridge to transplantation. Left ventricular assistance was maintained for up to 214 days (> 6 months), and renal and hepatic function were monitored at least weekly. RESULTS: Renal function before LVAD implantation in these patients was normal in 7 (creatinine, < 1.5) and moderately impaired in 3 (creatinine, 1.2 to 2.0). Hepatic function was normal in 7 patients before LVAD implantation (total bilirubin< 1.2; serum glutamic-oxaloacetic transaminase (SGOT), < 40; serum glutamic-pyruvic transaminase (SGPT), < 50) and normal at the time of transplantation in all 10 patients. Of the 3 patients with abnormal hepatic function before LVAD implantation, 1 patient had also had moderate renal dysfunction. CONCLUSIONS: Despite reduced pulsatility, the Jarvik 2000 LVAD improves or maintains excellent renal and hepatic function during periods of circulatory assistance in patients awaiting transplantation. PMID- 14530007 TI - Myosplint decreases wall stress without depressing function in the failing heart: a finite element model study. AB - BACKGROUND: The Myocor Myosplint is a transcavitary tensioning device designed to change left ventricular (LV) shape and reduce wall stress. Regional wall stress cannot be measured in the intact heart and LV function after surgical remodeling is often confounded by inotropic agents and mitral repair. We used a realistic mathematical (finite element) model of the dilated human LV to test the hypothesis that Myosplint decreased regional ventricular fiber stress and improved LV function. METHODS: A finite element model was used to simulate the effects of Myosplint on the LV stroke volume/end-diastolic pressure (Starling) relationship and regional distributions of stress in the local muscle fiber direction (fiber stress) for a wide range of diastolic and end-systolic material properties. The nonlinear stress-strain relationship for the diastolic myocardium was anisotropic with respect to the local muscle fiber direction. An elastance model for active fiber stress was incorporated in an axisymmetric geometric model of the globally dilated LV wall. RESULTS: Both diastolic compliance and end systolic elastance shifted to the left on the pressure-volume diagram. LV end diastolic volume and end-systolic volumes were reduced by 7.6% and 8.6%, respectively. Mean end-diastolic and end-systolic fiber stress was decreased by 24% and 16%, respectively. Although the effect of Myosplint on the Starling relationship was not significant, there were trends toward an improvement in this relationship at low diastolic stiffness, C, high peak intracellular calcium concentration, Ca(0), and high arterial elastance, E(A). Of note, the effect of C was twice that of Ca(0) and E(A). Diastolic function would, therefore, be expected to be the prime determinant of success with Myosplint. CONCLUSIONS: Myosplint reduces fiber stress without a decrement in the Starling relationship. Myosplint should be much more effective than partial ventriculectomy as a surgical therapy for patients with dilated cardiomyopathy and end-stage congestive heart failure. PMID- 14530009 TI - Brain protection using antegrade selective cerebral perfusion: a multicenter study. AB - BACKGROUND: To evaluate the results of antegrade selective cerebral perfusion as a method of brain protection during surgery of the thoracic aorta and to determine predictors of hospital mortality and adverse neurologic outcome. METHODS: Between October 1995 and March 2002, 588 patients underwent aortic surgery with the aid of antegrade selective cerebral perfusion. There were 334 men (56.8%); the mean age was 63.7 +/- 11.8 years. One hundred sixty-two patients (27.6%) underwent urgent operation. The separated graft technique was employed to reimplant the arch vessels in 230 patients (65.3%) of the 352 requiring aortic arch replacement. Associated procedures were performed in 254 patients (43.2%). One hundred twelve patients underwent elephant trunk procedure. The mean cerebral perfusion time was 67 +/- 37 minutes. RESULTS: The overall hospital mortality rate was 8.7%. A logistic regression analysis revealed urgent operation, recent central neurologic event, tamponade, unplanned coronary artery revascularization and pump time to be independent predictors of hospital mortality (p < 0.05). The permanent neurologic dysfunction rate was 3.8%. A logistic regression analysis showed tamponade to be independent predictor of permanent neurologic dysfunction (p < 0.05). The transient neurologic dysfunction rate was 5.6%. Recent central neurologic event, tamponade, coronary disease, and aortic valve replacement were indicated as independent predictors of transient neurologic dysfunction by logistic regression (p < 0.05). CONCLUSIONS: In our experience the utilization of antegrade selective cerebral perfusion resulted in encouraging results in terms of hospital mortality and brain complications. Neither the extent of the replacement nor the duration of the cerebral perfusion had an impact on hospital mortality and neurologic outcome. PMID- 14530010 TI - Safety of lumbar drains in thoracic aortic operations performed with extracorporeal circulation. AB - BACKGROUND: The safety of cerebrospinal fluid (CSF) drainage in thoracic aortic surgery using extracorporeal circulation (ECC) with systemic heparinization has not been established. METHODS: Four hundred thirty-two patients had descending thoracic or thoracoabdominal aortic repair between 1993 and 2002. One hundred sixty-two of those patients (age range, 67 +/- 13 years) had repairs performed with ECC, systemic anticoagulation, and lumbar CSF drainage. Repairs performed without CSF drainage, without ECC, or by stent graft (n = 53) were excluded. The CSF catheters were inserted at L3 to L5. Cerebrospinal fluid was drained to maintain pressures of 10 to 12 mm Hg. In the absence of neurologic deficit or coagulopathy, the catheters were capped at 24 hours and removed at 48 hours. Cerebrospinal fluid drainage was continued beyond 24 hours for delayed onset paraparesis. RESULTS: Cerebrospinal fluid drains were used in 135 thoracoabdominal aortic aneurysms (extent I, n = 63; extent II, n = 25; extent III, n = 39; extent IV, n = 8) and 27 descending thoracic aortic repairs (aneurysm, n = 24; traumatic aortic injury, n = 2; aortic coarctation, n = 1). Partial left heart bypass was used in 132 patients, full cardiopulmonary bypass without deep hypothermic circulatory arrest in 5, and cardiopulmonary bypass with adjunctive deep hypothermic circulatory arrest in 25. Time between catheter insertion and anticoagulation was 153 +/- 60 minutes. Heparin achieved an average maximum activated clotting time of 528 +/- 192 seconds. Average ECC time was 114 +/- 77 minutes. Average deep hypothermic circulatory arrest time was 40 +/- 12 minutes. Mortality was 14.1% (23 of 162), and permanent paraplegia was 4.9% (8 of 162). No epidural or spinal hematoma was observed. Six (3.7%) patients had catheter-related complications (temporary abducens nerve palsy [n = 1]; retained catheter fragments [n = 2]; retained catheter fragment and meningitis [n = 1]; isolated meningitis [n = 1]; and spinal headache [n = 1]). CONCLUSIONS: The CSF drainage in thoracic aortic surgery using ECC with full anticoagulation did not result in hemorrhagic complications. The permanent paraplegia rate in this complex patient population consisting of combined distal arch, thoracoabdominal aortic procedures were low, and lumbar CSF catheter-related complications had no permanent sequelae. PMID- 14530011 TI - The incidence of transient neurologic dysfunction after ascending aortic replacement with circulatory arrest. AB - BACKGROUND: Transient neurologic dysfunction (TND) namely postoperative confusion, delirium, and agitation after aortic operation, particularly after deep hypothermic circulatory arrest (DHCA), remains an underestimated adverse event in the early outcome of these patients. Although no influence on long-term outcome has been reported so far, this entity markedly affects the early outcome and leads to prolonged intensive care unit and hospital stay. METHODS: Between January 1997 and January 2003, 160 consecutive patients (130 type A dissections [81%] and 30 elective atherosclerotic aneurysms [19%]) had surgical repair with DHCA for a thoracic aortic aneurysm limited to the ascending aorta. From those, 40 patients (25%) underwent DHCA alone, whereas in 13 patients (8%) antegrade cerebral perfusion and in 103 patients (64%) retrograde cerebral perfusion was used for further brain protection. RESULTS: The overall incidence of TND was 18% (28 of 160) with a significant association between duration of circulatory arrest and the incidence of TND (13.8% in DHCA < 30 minutes versus 37.9% in DHCA > 40 minutes; p < 0.05). Furthermore the severity of TND was directly associated with the duration of circulatory arrest and age. In contrast, however, the use of retrograde cerebral perfusion had no influence on the incidence of TND, (p < 0.05). Intensive care unit stay as well as hospital stay were prolonged in the patients with TND (intensive care unit 14.3 +/- 14.2 days versus 10.8 +/- 13.7 days, p < 0.05; hospital stay 15.6 +/- 10.1 days versus 11.4 +/- 7.9 days, p < 0.05). CONCLUSIONS: Duration of DHCA, regardless of whether retrograde cerebral perfusion was used, was the most important predictor of the incidence of transient neurologic dysfunction in patients who had replacement of the ascending thoracic aorta. The occurrence of TND leads to impaired functional recovery as well as prolonged intensive care unit and hospital stay. PMID- 14530012 TI - Late reoperation for proximal aortic and arch complications after previous composite graft replacement in Marfan patients. AB - BACKGROUND: Marfan patients who received composite graft replacement for proximal aortic disease frequently require late reoperation. The initial surgical technique for this lesion remains controversial. METHODS: Fourteen Marfan patients who received composite graft replacement for annuloaortic ectasia with or without aortic dissection required late reoperation thorough re-median sternotomy. The techniques used for an initial composite graft replacement were the original Bentall procedure in 11 patients, the Cabrol procedure in 2, and coronary button technique in 1. Reoperation was indicated for prosthesis-related complications in 10 patients, distal aortic lesion in 13, or for both lesions in 8. Reoperations were performed, on average, 8.4 years after an initial operation. Reoperative procedures included re-composite graft replacement in 1 patient, total arch replacement in 5, and re-composite graft replacement with total arch replacement in 8. RESULTS: There were two in-hospital deaths (14.3%). Although pseudoaneurysms of the coronary artery or distal aorta occurred in the original Bentall or Cabrol procedures, true aneurysms of the coronary artery were noted even in the coronary button technique. Six patients required a total of eight subsequent descending or thoracoabdominal aortic replacements for an aneurysmal formation of a distal false lumen. CONCLUSIONS: The coronary button technique, with a small side hole for coronary anastomosis, is the procedure of choice for annuloaortic ectasia because it reduces the risk of coronary artery-related complications. Concomitant total arch replacement may be recommended for annuloaortic ectasia with DeBakey type I aortic dissection in selected patients to avoid the risk of reoperation on the aortic arch. PMID- 14530014 TI - Is extended arch replacement for acute type a aortic dissection an additional risk factor for mortality? AB - BACKGROUND: We report our experience with surgery for acute type A aortic dissection with involvement of the aortic arch. METHODS: From January 1986 to December 2001, 277 patients underwent surgery for acute type A aortic dissection. In 70 patients (25.3%), surgery was extended into the aortic arch: hemiarch and total arch replacement in 53 (75.7%) and 17 (24.3%) patients, respectively. Deep hypothermic circulatory arrest was used in 19 patients, antegrade selective cerebral perfusion in 38, and combined deep hypothermic circulatory arrest with antegrade selective cerebral perfusion in 13. RESULTS: Operative mortality was 18.6% (13/70) after extended replacement into the arch versus 21.7% (45/207) after surgery limited to the ascending aorta (p = 0.62). Multivariate analysis did not reveal significant risk factors for operative mortality. Postoperatively, 5 patients (8.1%) had a new postoperative cerebral vascular accident (CVA). Multivariate analysis showed an earlier date of operation as the only independent determinant for a new postoperative CVA (p = 0.0162, RR = 0.80/year, 95% CI = 0.67 to 0.96). None of the patients, operated on with antegrade selective cerebral perfusion, had a new cerebral deficit. Comparing the different methods of cerebral protection, multivariate risk analysis revealed antegrade selective cerebral perfusion as a significant protective factor against new postoperative CVA (p = 0.0110, OR = 0.12, 95% CI = 0.02 to 0.61). Survival at 5 and 10 years was 66.6.5% and 40.0%, respectively, after replacement of the aortic arch versus 68.7% and 57.7%, respectively, after replacement of the ascending aorta (p = 0.96). Freedom from aortic arch reoperation was 96.3% at 5 and 77.0% at 10 years versus 86.6% and 75.1% in both groups, respectively (p = 0.21). CONCLUSIONS: Extended replacement into the aortic arch during surgery for acute type A dissection does not influence early and late results. The best cerebral protection seems to be obtained with antegrade selective cerebral perfusion. PMID- 14530015 TI - pH-stat versus alpha-stat perfusion strategy during experimental hypothermic circulatory arrest: a microdialysis study. AB - BACKGROUND: The superiority of the pH-stat to the alpha-stat acid-base strategy during cardiopulmonary bypass as a neuroprotective method during hypothermic circulatory arrest is still controversial. In the present study, brain metabolism and outcome have been evaluated in a surviving model of experimental hypothermic circulatory arrest. METHODS: Twenty pigs undergoing 75-minutes of hypothermic circulatory arrest at a brain temperature of 18 degrees C were randomly assigned to the alpha-stat (n = 10) or pH-stat (n = 10) strategy during cardiopulmonary bypass. RESULTS: The 7-day survival rate was 90% (9 of 10) in the pH-stat group and 10% (1 of 10) in the alpha-stat group. At the end of cooling, pH-stat strategy was associated with significantly lower brain lactate and pyruvate concentrations and brain lactate-glucose ratio. After reperfusion, brain concentrations of glycerol, lactate, pyruvate, and lactate-glucose ratio were significantly lower in the pH-stat group. This strategy was associated with a faster rise of brain tissue temperature and reoxygenation on reperfusion, which is likely secondary to improved cerebral perfusion. CONCLUSIONS: During cardiopulmonary bypass before and after a period of hypothermic circulatory arrest, acid-base management according to the pH-stat principles seemed to be associated with less derangements in cerebral metabolism, lower intracranial pressures, and excellent behavioral recovery and survival outcome. Because there is strong evidence of the beneficial metabolic effects related to this method, further studies using an experimental model of combined HCA and embolic brain injury are required to exclude a possible increased risk of cerebral embolism associated with the pH-stat strategy. PMID- 14530016 TI - Myocardial protection with intermittent cold blood during aortic valve operation: antegrade versus retrograde delivery. AB - BACKGROUND: Intermittent antegrade cold blood cardioplegia is superior to warm blood cardioplegia in patients who have aortic valve operation. This study compared the cardioprotective efficacy of intermittent antegrade and retrograde cold blood cardioplegia with emphasis on metabolic stress in the left and right ventricles. METHODS: Thirty-nine patients who had elective aortic valve replacement were prospectively randomly selected to receive intermittent antegrade or retrograde cold blood cardioplegia. Left and right ventricular biopsies were collected 5 minutes after institution of cardiopulmonary bypass and 20 minutes after cross-clamp removal and were used to determine metabolic changes. Metabolites (adenine nucleotides, amino acids, and lactate) were measured using high-powered liquid chromatography and enzymatic techniques. Serial measurement of troponin I release was also used as a marker of myocardial injury. RESULTS: Preoperative characteristics were similar between groups. There was no in-hospital mortality, and no differences were observed in postoperative complications. Preischemic concentration of taurine was significantly higher in left ventricular biopsies, whereas adenosine triphosphate tended to be lower in the left ventricle. At reperfusion adenosine triphosphate levels were significantly lower than preischemic levels in right but not left ventricles irrespective of the route of delivery. The alanine-glutamate ratio was significantly elevated in both ventricles. Myocardial injury as assessed by troponin I release was also significantly increased in both groups. CONCLUSIONS: Retrograde and antegrade intermittent cold blood cardioplegic techniques are associated with suboptimal myocardial protection. Metabolic stress was more pronounced in the right than the left ventricle irrespective of the cardioplegic route of delivery used. PMID- 14530017 TI - ONO-6818, a novel, potent neutrophil elastase inhibitor, reduces inflammatory mediators during simulated extracorporeal circulation. AB - BACKGROUND: Among the serine proteases, neutrophil elastase is a powerful cytotoxic enzyme and plays a pivotal role in the inflammatory response associated with cardiopulmonary bypass. This study assesses the effects of the specific inhibition of neutrophil elastase by a novel, potent, low-molecular-weight neutrophil elastase inhibitor, ONO-6818. We hypothesized that ONO-6818 reduces inflammatory mediators and modulates adhesion molecules and the deformability of neutrophils during simulated extracorporeal circulation. METHODS: Simulated extracorporeal circulation was established by recirculating fresh heparinized (3.75 U/mL) human blood for 120 minutes in a membrane oxygenator and a roller pump with and without 1.0 micromol/L of ONO-6818 (n = 9 for control group, n = 7 for ONO-6818 group). The neutrophil adhesion molecules, CD11b and L-selectin, and the cytoplasmic F-actin of neutrophils were measured by flow cytometry. Neutrophil deformability was evaluated using simulated silicon microcapillaries. Neutrophil elastase, interleukin 8, and C5b-9 were measured using enzyme immunoassay. RESULTS: Neutrophil elastase levels were significantly lower in the ONO-6818 group. ONO-6818 significantly reduced interleukin 8 and C5b-9 production. ONO-6818 did not modulate changes of CD11b and L-selectin during recirculation. Cytoplasmic F-actin content and changes of neutrophil deformability did not significantly differ between the groups. CONCLUSIONS: Inhibition of neutrophil elastase activity with ONO-6818 reduces further interleukin 8 production and the formation of the complement membrane attack complex, and this results in a reduction of neutrophil elastase levels during simulated extracorporeal circulation. This study suggests that specific neutrophil elastase inhibition with ONO-6818 is a feasible therapeutic option to attenuate the exaggerated inflammatory response associated with cardiopulmonary bypass. PMID- 14530018 TI - Protection of myocardium by cyclosporin A and insulin: in vitro simulated ischemia study in human myocardium. AB - BACKGROUND: The efficacy of myocardial protection by cyclosporin A (CSA) and insulin was tested in human right atrial myocardial slices subjected to simulated ischemia and reoxygenation. METHODS: Slices of right atrial trabeculae were obtained from patients undergoing elective cardiac surgery. Trabeculae were incubated with oxygenated glucose containing phosphate buffered saline (O(2), G PBS). After 30 minutes of stabilization the sections were exposed to 90 minutes of simulated ischemia (N(2), PBS without glucose) followed by 90 minutes reoxygenation (O(2), G-PBS). Cyclosporin A (0.2 micromol/L) or insulin (5 mU/mL) was added during the stabilization period prior the ischemia. Cell viability was measured by using 3-[4.5 dimethylthiazol 2-yl]-2,5-diphenyltetrazolium bromide (MTT), which is cleaved by active mitochondrial dehydrogenases of living cells. RESULTS: The viability of untreated slices (control) was 30.45% +/- 2.5% versus 52.65% +/- 4.4% in the CSA treated slices, p less than 0.001. The extent of protection by CSA was affected by oral antiglycemic drugs (glibenclamide). The effect obtained by CSA was inhibited by 5-hydroxydecanoate (5HD), a specific blocker of mitochondrial K(ATP) channels. Protection of the myocardial slices with insulin appears to be superior and not affected by the medication before surgery. This protection was maximal when insulin was present during both preischemic equilibration and reoxygenation periods (68.9% +/- 9.3% viability with insulin versus 33.2% +/- 6.9% in the control, p < 0.001). CONCLUSIONS: Protection of right atrial trabeculae slices with insulin is superior to that obtained with CSA and is independent of preoperative medication. PMID- 14530019 TI - Comparison of developmental endothelial locus-1 angiogenic factor with vascular endothelial growth factor in a porcine model of cardiac ischemia. AB - BACKGROUND: This study compared the angiogenic effects of developmental endothelial locus-1 (DEL-1), vascular endothelial growth factor (VEGF), as well as the negative control, beta-galactosidase (beta-gal), in a porcine model of cardiac ischemia. METHODS: Twenty pigs underwent left circumflex artery occlusions. After 3 weeks, the animals received myocardial injections of adenovirus expressing beta-gal (n=6), DEL-1 (n=7), or VEGF (n=7). At 7 weeks, animals were assessed for both function and coronary flow and compared with baseline measurements. RESULTS: Regional wall motion index and global ejection fraction showed deterioration in function in the beta-gal group and no change in the VEGF and DEL-1 groups between the treatment and harvest time points. Preload recruitable stroke work suggested functional improvement in the VEGF group (35.8 +/- 8.6 vs 56.4 +/- 17.8, p = 0.033). The increase in the DEL-1 group was not statistically significant (27.3 +/- 9.8 vs, 40.2 +/- 19.4, p = 0.067). The beta gal group exhibited minimal change (30.7 +/- 14.8 vs 35.9 +/- 12.1, p = 0.96). Regional blood flow as assessed by fluorescent microspheres was improved under stress conditions in the VEGF group (1.00 +/- 0.15 vs 1.15 +/-0.22, p = 0.03). CONCLUSIONS: Treatment with VEGF led to a modest improvement in regional blood flow and cardiac function in previously ischemic myocardial tissue. PMID- 14530020 TI - Intra-myocyte ion homeostasis during ischemia-reperfusion injury: effects of pharmacologic preconditioning and controlled reperfusion. AB - BACKGROUND: This study determines whether controlled reperfusion or diazoxide improves intramyocyte Na(+) homeostasis using a porcine model of severe ischemia reperfusion injury. METHODS: Three groups (n = 10 pigs per group) had 75 minutes of left anterior descending artery occlusion during bypass. Group 1 had no treatment (control group), group 2 had controlled reperfusion (500 mL warm cardioplegia) (controlled reperfusion group), and group 3 had diazoxide (50 micromol/L before left anterior descending artery occlusion) (diazoxide group). Biopsies were taken from the left anterior descending artery region before ischemia and at 3, 5, and 10 minutes postreperfusion. Intra-myocyte Na(+) and water contents were determined using atomic absorption spectroscopy, and Na(+) concentrations were calculated. RESULTS: Intra-myocyte Na(+) increased for the diazoxide group pigs at 3-minutes postreperfusion (21.9 +/- 2.9 vs 34.0 +/- 3.4 micromol/mL; p = 0.02), but decreased to 19.9 +/- 3.2 micromol/mL at 10 minutes postreperfusion (p = 1.0 vs baseline). At 10 minutes postreperfusion, intra myocyte Na(+) in the controlled reperfusion group was lower than baseline (22.3 +/- 2.7 vs 17.2 +/- 3.1 micromol/mL; p < 0.001). Intra-myocyte Na(+) at 10 minutes postreperfusion for the diazoxide and controlled reperfusion groups was lower than for the control group (p < 0.05). CONCLUSIONS: Diazoxide and controlled reperfusion improved intra-myocyte Na(+) homeostasis after severe ischemia-reperfusion injury. PMID- 14530022 TI - Atrial compartment operation for atrial fibrillation: to isolate the left atrium or not? AB - BACKGROUND: The atrial compartment operation was designed to convert atrial fibrillation (AF) to sinus rhythm with intentional preservation of the electrical connection between adjacent atrial compartments. However, incidental left atrial isolation was observed in some patients. This study compared the long-term clinical outcomes of left atrial isolation for AF with those with right and left atrial connection. METHODS: Twenty patients with mitral valve disease and chronic AF who underwent atrial compartment operation with successful sinus conversion were studied. Left atrial isolation was documented by local electrogram recording. When there were no signs of electrical connection between the left atrium and the rest of the heart, either during sinus rhythm or during stimulation from various atrial compartments, left atrial isolation was confirmed. All patients were followed by electrocardiogram and echocardiogram serial recordings. Clinical signs and symptoms of cardiac performance and thromboembolism were also examined. RESULTS: Seven patients showed an isolated left atrium and 13 patients had electrical connection between the right and left atria. The age, gender, AF duration, and underlying disease were not different between the two groups of patients. During a mean follow-up period of 66 +/- 15 months, none of the patients with left atrial isolation showed recurrence of AF, although one experienced paroxysmal atrial flutter. However, 8 of the 13 patients with right and left atrial connection experienced recurrent atrial flutter/fibrillation (6 atrial flutter and 5 AF) (p = 0.058). The propensity for recurrent atrial flutter/fibrillation in these patients may be related to the conduction delay between the two atrial compartments, measured at 142 +/- 48 ms. At the end of the follow-up period, all patients with left atrial isolation remained in normal sinus rhythm without antiarrhythmic drugs. Of the patients who had right and left atrial connection, 2 developed sustained AF and 1 developed atrial flutter. Patients with left atrial isolation showed a decreased transmitral "A" flow compared with those with right and left atrial connection. Postoperative left atrial diameter and clinical functional class did not differ between patients with and without left atrial isolation. The incidence of embolization observed in both treatment groups did not differ significantly: 14% (1/7) in patients with left atrial isolation and 8% (1/13) in patients with right and left atrial connection (p > 0.05 between the groups). CONCLUSIONS: Left atrial isolation confers a better arrhythmia outcome but at the expense of poorer mechanical performance as compared with preserved electrical connection between the two atria. Nonetheless, all patients remain at risk for systemic embolization. Therefore, modifications of current surgical incisions for AF are needed. PMID- 14530023 TI - Cariporide is cardioprotective after iatrogenic ventricular fibrillation in the intact swine heart. AB - BACKGROUND: We sought to introduce sodium-hydrogen exchange inhibition as prophylaxis against the development of ventricular dysfunction in the setting of implantable cardioverter defibrillator insertion in high-risk patients. Cariporide, shown to be safe in humans, was used to reproduce previous results in our laboratory that demonstrated that sodium-hydrogen exchange inhibition preserves left ventricular (LV) function after ventricular fibrillation (VF) and reperfusion. METHODS: Twelve pigs (weight, 35 to 55 kg) were divided into two groups of six. Baseline ventricular function studies were based on echocardiography, conductance, aortic flow, and LV pressure. Animals were given vehicle (control) or cariporide (3 mg/kg intravenously). Ten minutes later, hearts underwent 80 seconds of VF. After reperfusion for 40 minutes, function studies were repeated. RESULTS: Postmortem examination included measuring passive pressure-volume curves and myocardial water content. Systolic indices, including preload recruitable stroke work and ejection fraction, were significantly depressed from baseline after VF and reperfusion for control animals (preload recruitable stroke work, 30.13 +/- 0.59 [standard error of the mean] versus 43.85 +/- 2.60 mm Hg; ejection fraction, 25.7% +/- 2.4% versus 33.5% +/- 3.0%) but not for those in the cariporide group (preload recruitable stroke work, 38.36 +/- 1.87 versus 40.86 +/- 1.45 mm Hg; ejection fraction, 33.9% +/- 3.5% versus 32.8% +/- 3.9%). In vivo diastolic indices demonstrated trends toward diminished ventricular compliance in control animals but not in the cariporide group after VF and reperfusion. Control animals had significantly increased postmortem LV stiffness, myocardial water content, and normalized LV mass. CONCLUSIONS: Cariporide preserves LV function after 80 seconds of VF and 40 minutes of reperfusion. Cariporide may prove useful in patients with severe LV dysfunction undergoing VF for implantable cardioverter defibrillator testing. PMID- 14530024 TI - Reducing the incidence and severity of pericardial adhesions with a sprayable polymeric matrix. AB - PURPOSE: This study was undertaken to evaluate the antiadhesive properties of a polymeric agent in infants undergoing staged surgical correction of congenital heart abnormalities. DESCRIPTION: Sixteen infants having staged surgical repair were treated with a polymeric matrix at the completion of the initial surgery. There were 5 untreated controls. The tenacity and extent of adhesions at five separate regions of the heart were evaluated at the follow-up surgery. EVALUATION: For all sites combined, there was a threefold difference in median tenacity scores in favor of the experimental treatment (1.0 vs 3.0, p < 0.01). Significant differences were achieved separately at the right ventricle and the anterior surface of the great vessels (p = 0.02 for both comparisons). Analysis of adhesion scores reflecting the extent of adhesions similarly favored the experimental treatment for all sites (80 vs 270, p < 0.01), with significant differences persisting at the right atrium (p < 0.01) and the anterior surface of the great vessels (p = 0.04). There were no treatment-related adverse events. CONCLUSIONS: Use of this polymeric agent at the completion of open cardiac surgery may prevent the occurrence or reduce the severity of pericardial adhesions. PMID- 14530026 TI - A pulmonary cavernous hemangioma causing massive hemoptysis. AB - A 54-year-old man was seen with massive hemoptysis. A posteroanterior chest radiograph revealed increased bronchovascular branching in the left hemithorax. Thoracic computed tomography showed a well-demarcated mass with uniform density confined to the apicoposterior and anterior segments of the left upper lobe. Fiberoptic bronchoscopy revealed a locus of bleeding in the apicoposterior segment of that lobe. Exploratory left thoracotomy confirmed the presence of a mass. The results of frozen section examination of a biopsy specimen were benign. The mass was resected by upper lobectomy. The definitive result of histopathological study of the mass was cavernous hemangioma. Very few cases of pulmonary hemangioma have been reported in the literature. PMID- 14530027 TI - Weerda diverticuloscope: novel use to remove embedded esophageal foreign bodies. AB - Embedded sharp foreign bodies of the cervical esophagus represent a clinical challenge. Initial attempts at removal are usually undertaken by nonsurgical endoscopists who are often successful with blunt objects. Unsuccessful attempts with sharp objects, however, can result in distal migration, mucosal damage, and frank perforation. Thoracic surgeons are often called on for cervical esophagotomy after endoscopic attempts have failed. This report describes the novel use of a Weerda diverticuloscope for removal of a dental appliance with metallic hooks embedded in the cervical esophagus. PMID- 14530028 TI - Papillary adenocarcinoma in a thymic cyst: a pitfall of thoracoscopic excision. AB - Carcinoma arising in a thymic cyst is very rare. We performed thoracoscopic subtotal resection of an assumed benign cyst. The patient had a reoperation when the diagnosis of papillary adenocarcinoma was established. Complete resection and pleural thermochemotherapy were performed. The patient died 26 months later from systemic metastases. Caution must be exercised when managing nontypical thymic cysts. PMID- 14530029 TI - Surgery for unilateral absence of pulmonary artery using autologous tissue. AB - A 7-year-old girl with unilateral absence of the pulmonary artery underwent autologous tissue limited reconstructive surgery. The proximal portion of the artery was reconstructed by rotating a reverse U-shaped cut opposite the pulmonary arterial wall and covering the anterior surface with autologous pericardium. Follow-up catheterization at 5.8 years after surgery revealed no stenosis. This procedure could become one of the preferred methods for this unusual clinical condition. PMID- 14530030 TI - Staged biventricular repair of Taussig-Bing anomaly with subaortic stenosis and coarctation of aorta. AB - We present successful procedures for 2 infants who had the Taussig-Bing anomaly with subaortic stenosis and coarctation of the aorta. The initial procedure was coarctoplasty and the Damus-Kaye-Stansel procedure with modified Blalock-Taussig shunt. The second procedure was intraventricular repair (Kawashima procedure), Damus-Kaye-Stansel take-down and the reuse of native aortic and pulmonary valves 19 and 25 months later. Both patients survived the operations and postoperative hemodynamics were excellent at both 28 and 59 months follow-ups. PMID- 14530032 TI - Reimplantation of anomalous right coronary artery arising from the pulmonary trunk leading to normal coronary flow reserve late after surgery. AB - We report a case of successful reimplantation of the right coronary artery from the pulmonary trunk into the aorta with a 10-year follow-up. The finding of a normal coronary flow reserve late after surgery suggests that direct reimplantation of the right coronary artery into the aorta is the preferable surgical technique. PMID- 14530031 TI - Successful treatment for intraoperatively evolving acute aortic dissection in a neonate. AB - Acute aortic dissection is a life-threatening condition. We report the case of a neonatal one-stage correction of coarctation and hypoplastic aortic arch repair plus ventricular septal defect closure. While dissecting the head vessels after cannulation of the ascending aorta and commencing cardiopulmonary bypass, type A aortic dissection evolved. This required immediate ascending aorta and aortic arch reconstruction with coarctation of the aorta resection under hypothermic circulatory arrest. The surgical management of this rather unique situation is discussed here. PMID- 14530033 TI - Orthotopic cardiac transplantation 30 months after successful dynamic cardiomyoplasty. AB - Reports of cardiac transplantation after successful cardiomyoplasty are rare. We report the case of a 63-year-old man with intractable heart failure who underwent successful orthotopic cardiac transplantation 30 months after dynamic cardiomyoplasty. PMID- 14530035 TI - Thrombolytic therapy for delayed, in-hospital stroke after cardiac surgery. AB - Five patients who had delayed stroke after cardiac surgery underwent intraarterial administration of a fibrinolytic agent for thromboembolism (n = 4) or thrombosis (n = 1) of the cerebral artery. Complete recanalization of the occluded artery was obtained in 3 patients and partial recanalization in 2. Additional angioplasty for basilar artery stenosis was performed in 1 patient. No patients exhibited rebleeding into the pericardial space or wound bleeding. All patients survived with moderate or full functional recovery. Immediate cerebral angiography and local thrombolysis may improve functional outcome and survival in patients with postcardiotomy cerebral thromboembolism. PMID- 14530034 TI - Circumflex artery stenosis induced by intraoperative radiofrequency ablation. AB - We report a case of circumflex artery stenosis after intraoperative radiofrequency ablation for permanent atrial fibrillation in a patient who had a previous mitral valve replacement. The patient presented with acute pulmonary edema and severe angina 1 year after an uneventful recovery. The patient underwent a diagnostic angiography that showed the presence of stenosis of a long segment of the circumflex artery, adjacent to the radiofrequency ablation site, which was reopened successfully by angioplasty. Intraoperative radiofrequency ablation caused circumflex artery stenosis. We believe that this complication could have been avoided by applying the radiofrequency ablation more distally between the left pulmonary veins and the mitral valve. PMID- 14530036 TI - Aortic arch homograft as a bypass conduit for superior vena cava obstruction. AB - Superior vena cava (SVC) obstruction most commonly results from malignant disease of the superior mediastinum, which is amenable to percutaneous stenting. Superior vena cava syndrome can also be caused by transvenous pacemaker electrodes and indwelling venous catheters, when it may be refractory to minimally invasive treatment. We report 2 patients with superior vena cava obstruction treated successfully by a surgical bypass approach using cryopreserved aortic arch homografts. PMID- 14530037 TI - Preoperative construction of an extracranial arterial shunt for resection of an aortic arch aneurysm with occluded left carotid artery. AB - A 68-year-old man with aortic arch aneurysm was referred to our department. Preoperative carotid echography and magnetic resonance angiography revealed occlusion of the left internal carotid artery. Single-photon emission computed tomography scanning indicated that cerebral blood flow was decreased and reactivity to acetazolamide was reduced in the left temporal lobe. A successful superficial temporal artery-middle cerebral artery anastomosis was first made by neurosurgeons. A postoperative single-photon emission computed tomography scan showed that cerebral blood flow and reactivity to acetazolamide were remarkably improved. Two months after the anastomosis, the aortic arch aneurysm was successfully repaired. PMID- 14530038 TI - Aortic dissection coexistent with two saccular aneurysms. AB - We report a rare case of DeBakey type I aortic dissection coexistent with two saccular aneurysms adjacent but not communicating with the aortic dissection. PMID- 14530039 TI - Infectious endocarditis of a Chiari network. AB - We present the case of a 62-year-old man with infectious endocarditis in a Chiari network. Chiari networks are present in 1.5% to 3% of the population. Although Chiari networks are usually clinically insignificant, they are associated with a number of conditions, including patent foramen ovale, thromboembolism, atrial aneurysm, and cardiac arrhythmias. Although there are rare reports of patients with a Chiari network who had endocarditis develop, this is the first report of a patient who had endocarditis develop solely within a Chiari network. PMID- 14530040 TI - Successful resection of a symptomatic right ventricular lipoma. AB - We report the case of a 31-year-old woman with a 4-year history of recurrent palpitations, presenting with an increased frequency of paroxysms caused by ventricular tachycardias during pregnancy. A cardiac tumor of unknown origin infiltrating the right ventricle was diagnosed. Three weeks after prophylactic abrasion the tumor was totally excised with the use of cardiopulmonary bypass including restoration of the right ventricular wall and the tricuspid valve. Histology confirmed diagnosis of a benign cardiac lipoma. The postoperative course was uneventful and the patient was discharged 7 days after surgery. There was no episode of ventricular tachycardias during the 6-month follow-up. PMID- 14530041 TI - MDCT scan visualizes the Adamkiewicz artery. PMID- 14530042 TI - Giant postdissection aneurysm of the ascending aorta after aortic valve replacement. PMID- 14530043 TI - Thoracoscopic thymectomy using anterior chest wall lifting method. AB - We performed a thoracoscopic resection of thymus and thymoma using a novel method whereby the chest wall was lifted by costal hooks placed on the bilateral third ribs. Since the thymus and fat tissue were also elevated, the mediastinal dissection was started at the underlying vessels and pericardium. This technique conforms to the purpose of endoscopic surgery, as it maximizes the operative field and minimizes chest wall trauma. We have applied this approach to benign mediastinal tumor and stage I thymoma with satisfactory results. PMID- 14530044 TI - A technique for aortic valve replacement on the beating heart with continuous retrograde coronary sinus perfusion with warm oxygenated blood. AB - The protection of ventricular myocardium in aortic valve operations is always an issue because those hearts do not tolerate global ischemia well. A technique of aortic valve replacement is described involving continuous retrograde coronary sinus perfusion with warm oxygenated blood used in 34 patients to date without any complications. This technique maintains a beating heart throughout the procedure. PMID- 14530046 TI - Warning hint of a hidden early left main stem bifurcation during antegrade cardioplegia. AB - The short left main trunk with early bifurcation is a common variation of the left coronary anatomy and is easily overlooked during antegrade selective cardioplegia resulting in the risk of single branch perfusion. We describe an obvious characteristic sign to detect this pitfall during blood cardioplegia. PMID- 14530045 TI - Homemade mitral ring. AB - A simple and quick technique to fashion a mitral ring was developed using a stainless steel wire covered by a Gore-Tex strip. This technique allows a 15-fold reduction in the cost of a mitral ring compared with commercially available rings. As such, cardiac surgery becomes more readily available to patients in developing countries such as Africa and Asia. These homemade rings were implanted in 6 patients with similar results to commercially available rings. PMID- 14530047 TI - Active, specific immunotherapy for lung cancer: hurdles and strategies using genetic modification. AB - Active immunotherapy for lung cancer has been a challenge because of the poor antigenic characterization of these tumors and their ability to escape the immune response. However, knowledge of the mechanisms of anti-tumor immunity has expanded significantly over the past decade, leading to the development of more novel, specific strategies for augmenting the immune response. Genetic manipulation of tumor cells, immune cells, or both, may help overcome some of the previously encountered difficulties of immunotherapy. Laboratory and clinical investigations are currently ongoing to evaluate the feasibility and potential benefit of these novel approaches. PMID- 14530048 TI - Lung cancer and cyclooxygenase-2. AB - Lung cancer is by far the leading cause of cancer-related death. Overall survival is poor and has not improved substantially over the last half century. It is clear that new approaches are needed and these should include prevention, screening for early detection, and novel treatments based on our understanding of the molecular biology of this disease. Recently attention has been drawn to the role of the cyclooxygenase (COX) enzyme and its involvement in tumorigenesis. Investigations have documented two isoforms, COX-1 and COX-2, encoded by different genes. COX-1 is constitutively expressed in most tissues and appears to be responsible for the production of prostaglandins mediating normal physiologic functions, such as the maintenance of gastric mucosa and regulation of renal blood flow. In contrast, COX-2 is normally undetectable in most tissues, and is induced by cytokines, growth factors, oncogenes, and tumor promoters. A growing body of evidence indicates COX-2 plays a key role in lung cancer, and can serve as a potential marker of prognosis in this disease. Furthermore, the recent availability of COX-2 inhibitor medications offers a unique opportunity to interfere with the development of lung cancer and the progression of metastasis. Because COX-2 inhibitors have been demonstrated to interfere with tumorigenesis, the COX-2 enzyme may be an attractive target for therapeutic and chemoprotective strategies in lung cancer patients. PMID- 14530049 TI - Legal consequences of disregarding the wishes of a patient. PMID- 14530052 TI - Continuous retrograde cardioplegia simplifies aortic valve replacement in the presence of a patent internal mammary artery. PMID- 14530055 TI - Randomized trial of stentless versus stented bioprostheses for aortic valve replacement. PMID- 14530056 TI - Are composite Y-grafts able to fully respond to the left coronary system flow demand early after coronary bypass graft? PMID- 14530057 TI - Early ischemic preconditioning for spinal cord protection. PMID- 14530062 TI - Understanding disparities in outcomes in cardiovascular medicine and thoracic oncology in African-American patients. PMID- 14530063 TI - Historical perspectives on the care of African Americans with cardiovascular disease. PMID- 14530064 TI - Cardiac surgery in African Americans. AB - Retrospective and prospective randomized studies that provide information on the influence of race on the morbidity and mortality of cardiac surgical procedures are reviewed. We intentionally focus our attention on the specific outcomes of these procedures in African Americans because African Americans have a high incidence of all-cause cardiovascular mortality and a high prevalence of a number of risk factors associated with cardiovascular mortality. Furthermore, numerous studies have confirmed that blacks, as a function of race, lack equal access to diagnostic and therapeutic invasive cardiac procedures. Here we use the terms "black" and "African American" interchangeably. In this context we interpret both terms to refer to Americans of African descent. Similarly, we use the term "white" or "Caucasian" interchangeably to refer to Americans of European descent. PMID- 14530065 TI - Lung cancer in African Americans. PMID- 14530066 TI - Geographical distribution and racial disparity in esophageal cancer. PMID- 14530067 TI - Impact of renal disease in cardiovascular surgery: emphasis on the African American patient. AB - Cardiovascular disease remains a significant source of morbidity and mortality for patients with kidney disease. Coincident with the development of chronic renal failure, patients typically manifest a systemic vasculopathy often involving the cardiovascular system. The renal failure patient is also plagued by multiple comorbid conditions that may adversely affect cardiovascular outcomes. Consistent with the national trend of increasing numbers of patients requiring renal replacement therapy (RRT), patients requiring invasive cardiovascular procedures are also on the incline. The morbidity and mortality related to these procedures has remained high despite significant advances in delivery and maintenance of care. Is the African-American patient with renal failure unique in terms of cardiovascular morbidity and mortality? Numerous studies have documented racial differences in access to invasive cardiovascular procedures, even after controlling for multiple physiologic risk factors and socioeconomic and sociocultural factors. Studies have also shown higher morbidity and lower survival for African-American patients after cardiac procedures. In this high risk population these same issues perhaps would persist. The following paper will examine the current status of cardiovascular disease in the renal failure patient with emphasis on the African-American patient population. PMID- 14530068 TI - Unequal treatment: report of the Institute of Medicine on racial and ethnic disparities in healthcare. PMID- 14530069 TI - On the influence of flow conditions and wettability on blood material interactions. AB - In this review, we hypothesise that, next to biocompatibility, optimal blood compatibility depends on a combination of biomaterials wettability and the shear stress prevailing in the device. The wettability is discussed in seven different categories of devices, that differ substantially from each other with regard to shear stress and exposure time. These seven categories are stents, prosthetic heart valves, vascular prostheses, cardiopulmonary bypass, hemodialysis, vena cava filters and blood bags. In high shear applications, in combination with blood activation, platelet deposition and thrombosis appear to be major problems and platelet inhibitors are most effective. Exposure of blood to a large biomaterial surface, with or without antithrombotic coating, results in reduction of platelet function. Material-independent activation aggravates this process. In low shear applications, platelets only seem supportive for coagulation and anticoagulants should be used. PMID- 14530070 TI - A model for the preliminary biological screening of potential keratoprosthetic biomaterials. AB - A series of in vitro screening assays for the preliminary selection of biomaterials for use in the fabrication of artificial corneas (keratoprostheses) (KPros) have been investigated. These screening assays assessed the initial binding of inflammatory and cell adhesive proteins, activation of inflammatory proteins, adhesion of keratocytes, epithelial cells and macrophages and the production of inflammatory cytokines by keratocytes contacting biomaterials. Central optic biomaterials were selected on the basis of low-inflammatory and cell adhesion potential. Peripheral skirt materials were selected on the basis of low-inflammatory potential but good cell adhesion to anchor the implant within the host cornea. Green fluorescent protein (GFP) gene transfer was used in a novel context to investigate cell invasion in the absence of external staining techniques. Confocal laser scanning microscopy and scanning electron microscopy were used to investigate GFP positive keratocyte invasion of porous materials. The results of in vitro assays were compared to a corneal organ culture system in which the biomaterials were assessed within a stromal environment. A range of polyurethane-based interpenetrating polymers with a range of water contents were screened. All materials showed low-inflammatory potential. A reduction in biomaterial water content induced an increase in complement C3 and fibronectin binding and in cell adhesion to materials, whilst differences in co-monomer formulation had little impact. The screening methods used in the current study provide a suitable preliminary assessment regime for the in vitro evaluation of potential KPro materials. PMID- 14530071 TI - Dissolution and mineralization behaviors of HA coatings. AB - The dissolution and mineralization behavior of HA coatings are two of the main factors governing the bioactivity of coatings. After different post treatment operators, the plasma-sprayed HA coatings have different characteristics, including different chemical composition, crystallinity, crystallite size and dissolution behavior. In this study, HA coatings were characterized by X-ray diffraction, scanning electron microscope, and Fourier transform infrared spectra before and after immersion in simulated body fluid (SBF). When immersed in SBF, both dissolution and precipitation occurred at the same time, but the kinetics of dissolution was quite different from that of precipitation. The former was dominated by ion exchange, while the latter was controlled by the ion concentration product and the solubility of the particles. Therefore, the dissolution behaviors of phosphate ions partly depended on the dissolution behaviors of calcium ions. With the increase of ions concentration in solution by dissolution, more nucleation sites appeared on the surface of coatings. Crystalline grains gradually grew up on the nucleation sites and developed into biomineral layers. The biomineral layers were the results of the precipitation of the ions in the solution; and the carbonates partially substituted phosphates to form bone-like apatite. The different dissolution characters resulted in quite different morphology of the biomineral layers: the coatings with low solubility induced biomineral layers of large grains; on the contrary, the biomineral layers of network structure were observed on the more soluble coatings. PMID- 14530072 TI - Influence of sterilization on the mineralization of titanium implants induced by incubation in various biological model fluids. AB - The aim of this work was to investigate the effect of the sterilization processes on the mineralization of titanium implants induced by incubation in various biological model fluids. Titanium samples were submitted to the following sterilization processes used for implant materials: steam autoclaving, glow discharge Ar plasma treatment and gamma-irradiation. The modification of the treated surfaces was evaluated by contact angle determinations, X-ray photoelectron spectroscopy (XPS), laser profilometry and X-ray diffraction. The most significant modifications were detected on the wettability: while the samples treated with Ar plasma became highly hydrophilic (water contact angle approximately 0 degrees), gamma-irradiation and steam sterilization induced an increase in the hydrophobicity. After being sterilized, the samples were incubated for one week in three biological model fluids: Hanks' Balanced Salt Solution, Kokubo's simulated body fluid (SBF) and a fluid, designated by SBF0, with the same composition of SBF but without buffer TRIS. The level of mineralization of the incubated Ti samples, assessed by dynamic contact angle analysis, scanning electron microscopy, electron dispersive spectroscopy and XPS, indicated that the early stages of mineralization are essentially independent of the sterilization method. In contrast, the incubating fluid plays a determinant role, SBFO being the most efficient medium for biomineralization of titanium. PMID- 14530073 TI - Chondrocyte interactions with porous titanium alloy and calcium polyphosphate substrates. AB - Chondrocytes maintain their phenotype and form cartilagenous tissue when cultured on calcium polyphosphate (CPP) or titanium alloy (Ti alloy), porous three dimensional materials. To understand how these materials may influence chondrocyte phenotype and matrix synthesis, the early interactions of cultured cells with CPP and titanium alloy were examined. These were compared to chondrocytes grown in monolayer culture on tissue culture polystyrene, conditions in which cultured chondrocytes dedifferentiate and do not form cartilagenous tissue. Scanning electron microscopy of cells up to 72 h in culture showed that bovine chondrocytes on CPP, Ti alloy, and polystyrene were an admixture of round and spread cells. The spread cells on CPP and titanium alloy were not entirely flattened but maintained a polygonal shape. In contrast, spread chondrocytes in monolayer culture were flatter and significantly larger, a difference that was maintained even in the absence of serum. All cells cultured on CPP and Ti alloy exhibited subcortical ring-like distribution of actin filaments whereas the flattened cells on polystyrene showed actin filaments distributed throughout the cytoplasm. Cells on CPP and Ti alloy synthesized significantly less collagen and proteoglycans than cells cultured on polystyrene at 72 h of culture. In summary the cells on the porous three-dimensional materials differed from those on polystyrene in terms of cell morphology and size, actin cytoskeleton organization, and synthesis of selected matrix macromolecules. The data suggests that CPP and titanium alloy may mediate their effect by limiting cell spreading in part by favoring the maintenance of a ring-like actin distribution. PMID- 14530074 TI - Enhanced DNA synthesis accompanied by constitutive phosphorylation of the ERK pathway in human fibroblasts cultured on a polyelectrolyte complex. AB - In this study, we examined the cellular and molecular responses of fibroblasts cultured on a polyelectrolyte complex (PEC) derived from sulfated chitin as a polyanion and chitosan as a polycation. On PEC-coated dishes, the fibroblasts aggregated and then developed spheroid-like structures. At earlier stages of culture, DNA synthesis of cells cultured on PEC was stimulated approximately 75% higher than control cells. Among various signaling molecules examined, including mitogen-activated protein kinases, Akt/PKB and p53, an extracellular-signal regulated kinase (ERK) was selectively and constitutively phosphorylated in cells cultured on PEC. The constitutive phosphorylation of ERK was derived from an activation of the ERK kinase MEK, but not from an inactivation of the ERK phosphatase MKP-1. Furthermore, ERK phosphorylation was almost abolished by a membrane receptor tyrosine kinase inhibitor. The enhanced phosphorylation of focal adhesion kinase, a downstream molecule of integrins, was also observed in cells cultured on PEC. These results suggest that fibroblasts recognize PEC as a continuous mitogenic stimulant which results in the constitutive activation of the MEK-ERK pathway toward mitogenesis. Further, PEC interacts with the cell membrane leading to activation of membrane molecules, including integrins and receptor tyrosine kinases. These responses may account, at least in part, for the potential use of PEC as a biomaterial for tissue regeneration. PMID- 14530075 TI - Properties of carbodiimide treated heparin. AB - Heparin, a well-known anticoagulant, has been frequently used to coat surfaces for attaining blood compatibility of polymeric materials. Since carbodiimides are often used for immobilization of heparin on these biomaterials, the present study intended to evaluate some properties of carbodiimide-treated heparin. It was observed that the properties of heparin were altered after treatment with carbodiimide, mainly in the molar excess of this reagent. Thus, dye fixation and electrophoretic behaviour of heparin were modified, as well as its degradability by specific enzymes. Also, these modifications resulted in loss of anticoagulant activity. Infrared spectra of this carbodiimide-treated heparin presented evidence that can confirm its modification. PMID- 14530076 TI - Semi-quantitative analysis of cytokines in MM THR tissues and their relationship to metal particles. AB - Cytokines associated with osteolysis have been demonstrated in tissues surrounding failed metal-metal (MM) total hip replacements (THRs). The objective of the present study was to semi-quantify the amounts of inflammatory cytokines in tissues from 28 failed MM THRs, and determine their relationship with the quantity of metal particles. Paraffin sections were immunohistochemically stained with monoclonal antibodies: anti-IL-1-beta, anti-IL-6 and anti-TNF-alpha. Cytokines and metal particles were rated in 10 fields per tissue using standard light microscopy. Because of the use of light microscopy, only relatively large particles or agglomerations of particles were visible. Therefore, a polarized light and a semi-quantitative scheme based on the discoloration of cell cytoplasms induced by the presence of particles were used to evaluate the quantity of metal particles. Results showed an overall higher amount of IL-6 than IL-1beta while TNF-alpha remained at very low levels. For each patient, the average IL-1beta and IL-6 ratings decreased when the average particle rating increased, following a linear regression, with relatively high correlation factors (r=-0.69 for IL-1beta and r=-0.57 for IL-6). IL-1beta decreased about twice as fast as IL-6. TNF-alpha, remaining at very low levels, did not demonstrate any correlation with particle rating. When multiple tissues were available for the same patient, the correlation factors between the average cytokine and particle ratings were highly variable between samples, demonstrating the heterogeneity between the tissues from the same patient. At the cellular level, there was an even higher correlation between the quantity of metal particles and the production of IL-1beta and IL-6 (r=-0.99), while TNF-alpha did not demonstrate any correlation, remaining at very low levels. In conclusion, this study showed that tissues surrounding failed MM THRs with low to moderate quantities of metal particles can induce the production of potentially osteolytic cytokines. However, the overall number of cells producing these cytokines tended to be lower than that typically seen in tissues surrounding metal-polyethylene THRs. PMID- 14530077 TI - Quantitative assessment of the response of primary derived human osteoblasts and macrophages to a range of nanotopography surfaces in a single culture model in vitro. AB - The effect of nanotopography on a range of Ti oxide surfaces was determined. Flat Ti, 3%, 19%, 30% and 43% topography densities of 110 nm high hemispherical protrusions were cultured in contact with primary derived human macrophages and osteoblasts in single culture models. Prior to introduction of the test substrate the phenotype and optimum conditions for in vitro cell culture were established. The cellular response was investigated and quantified by assessments of cytoskeletal development and orientation, viable cell adhesion, cytokine production and release and RT-PCR analysis of osteogenic markers. The tested nanotopographies did not have a statistically significant effect on viable cell adhesion and subsequent cytoskeletal formation. Surface chemistry was the dominant factor as established via incorporation of a tissue culture polystyrene, TCPS, control. The topography surfaces induced a release of chemotactic macrophage activation agents at 1 day in conjunction with stress fibre formation and a subsequent fibronectin network formation. Osteoblasts migrated away from the topography surfaces to the exposed TCPS within the wells during the 7-day period. PMID- 14530078 TI - Distinct gene expression of receptor activator of nuclear factor-kappaB and rank ligand in the inflammatory response to variant morphologies of UHMWPE particles. AB - Recent studies have examined the role of wear debris-induced bone resorption in the aseptic loosening of orthopedic prostheses. Research has shown that inflammation depends not only on the amount of particulate debris, but also the shape and size of the accumulated wear particles. Our previous studies have demonstrated that variant shapes of ultra-high molecular weight polyethylene (UHMWPE) particles induce diverse cellular and apoptotic responses in a murine inflammation model. Since enhanced osteoclastogenesis is recognized as a hallmark of bone loss in prosthetic loosening, we have now investigated the gene expression of receptor activator of nuclear factor-kappaB (RANK) and receptor activator of nuclear factor-kappaB ligand (RANKL) during the inflammatory response to different shapes of UHMWPE particles. Two shapes of UHMWPE particles (globular or elongated) were implanted in established air pouches on BALB/c mice, and pouches harvested 7 days after stimulation with UHMWPE particles. Gene levels of RANK, RANKL, TNFalpha, IL-1beta, and cathepsin K (CK) were quantified by real time RT-PCR, and TRAP staining of pouch membrane was used to evaluate osteoclastogenesis. We found that (i) elongated particles generated significantly higher RANK and RANKL gene expression than globular particles in pouch tissue; (ii) elongated particles provoked significantly higher IL-1beta and TNFalpha gene expression; (iii) a positive association was found between tissue inflammation status and the gene level of RANK/RANKL; and (iv) elongated particles stimulated significantly higher CK gene expression in comparison with globular particles. Histology revealed that clusters of TRAP+ cells were located in regions in contact with elongated particles. Overall, these data suggest that the morphology of wear debris may be a critical factor in the pathogenesis of prosthetic loosening. PMID- 14530079 TI - New biomorphic SiC ceramics coated with bioactive glass for biomedical applications. AB - A new generation of light, tough and high-strength material for medical implants for bone substitutions with a good biological response is presented. The innovative product that fulfills all these requirements is based on biomorphic silicon carbide ceramics coated with a bioactive glass layer. The combination of the excellent mechanical properties and low density of the biomorphic SiC ceramics, used as a base material for implants, with the osteoconducting properties of the bioactive glass materials opens new possibilities for the development of alternative dental and orthopedic implants with enhanced mechanical and biochemical properties that ensures optimum fixation to living tissue. Biomorphic SiC is fabricated by molten-Si infiltration of carbon templates obtained by controlled pyrolysis of wood. Through this process, the microstructure of the final SiC product mimics that of the starting wood, which has been perfected by natural evolution. The basic features of such microstructure are its porosity (ranging from 30% to 70%) and its anisotropy, which resembles the cellular microstructure and the mechanical characteristics of the bone. The SiC ceramics have been successfully coated with a uniform and adherent bioactive glass film by pulsed laser ablation using an excimer ArF laser. The excellent coverage of the SiC rough surface without film spallation or detachment is demonstrated. In order to assess the coating bioactivity, in vitro tests by soaking the samples in simulated body fluid have been carried out. After 72 h, the formation of a dense apatite layer has been observed even in interconnecting pores by SEM and energy dispersive X-ray spectroscopy analysis demonstrating the bioactive response of this product. PMID- 14530080 TI - Collagen/chitosan porous scaffolds with improved biostability for skin tissue engineering. AB - Porous scaffolds for skin tissue engineering were fabricated by freeze-drying the mixture of collagen and chitosan solutions. Glutaraldehyde (GA) was used to treat the scaffolds to improve their biostability. Confocal laser scanning microscopy observation confirmed the even distribution of these two constituent materials in the scaffold. The GA concentrations have a slight effect on the cross-section morphology and the swelling ratios of the cross-linked scaffolds. The collagenase digestion test proved that the presence of chitosan can obviously improve the biostability of the collagen/chitosan scaffold under the GA treatment, where chitosan might function as a cross-linking bridge. A detail investigation found that a steady increase of the biostability of the collagen/chitosan scaffold was achieved when GA concentration was lower than 0.1%, then was less influenced at a still higher GA concentration up to 0.25%. In vitro culture of human dermal fibroblasts proved that the GA-treated scaffold could retain the original good cytocompatibility of collagen to effectively accelerate cell infiltration and proliferation. In vivo animal tests further revealed that the scaffold could sufficiently support and accelerate the fibroblasts infiltration from the surrounding tissue. Immunohistochemistry analysis of the scaffold embedded for 28 days indicated that the biodegradation of the 0.25% GA-treated scaffold is a long term process. All these results suggest that collagen/chitosan scaffold cross linked by GA is a potential candidate for dermal equivalent with enhanced biostability and good biocompatibility. PMID- 14530081 TI - Insulinotropic activity of sulfonylurea/pullulan conjugate in rat islet microcapsule. AB - The in vitro long-term effect of a water-soluble sulfonylurea/pullulan conjugate (SUP) on insulinotropic activity and cell viability was investigated using rat pancreatic islets co-entrapped with SUP in conventional alginate-poly(L-lysine) microcapsules. The conjugate was synthesized by coupling a carboxylated glibenclamide derivative to a polysaccharide, pullulan (MW=200,000). In vitro static experiment showed that sulfonylurea concentration in SUP over 50 microM was required to stimulate the rat islets. In a dynamic insulin secretion test, the microcapsules of islets with SUP regained the insulin secretion pattern comparable to that of free islets, while those without SUP showed impaired insulin secretion. The long-term (1 month) culture experiment demonstrated that the microcapsules of islets with SUP, with well-preserved morphology, presented higher insulin secretion level and better ability in responding to glucose changes than those without SUP. PMID- 14530082 TI - Gelatin-chondroitin-hyaluronan tri-copolymer scaffold for cartilage tissue engineering. AB - The mechanism by which the cell synthesizes and secretes extracellular matrix (ECM) and is, in turn, regulated by the ECM is termed dynamic reciprocity. The aim of the present work was to produce a gelatin/chondoitin-6-sulfate/hyaluronan tri-copolymer to mimic natural cartilage matrix for use as a scaffold for cartilage tissue engineering. The scaffold produced had a uniform pore size of about 180 microm and adequate porosity of 75%. Porcine chondrocytes were seeded onto the tri-copolymer scaffold and cultured in Petri dishes or spinner flasks for 2, 3, 4, or 5 weeks. Chondrocytes were uniformly distributed in the scaffold in the spinner flask cultures, but less so in the Petri dish cultures. Secretion of ECM was found under histology examination. In spinner flask cultures, chondrocytes retained their phenotype for at least 5 weeks, as shown immunohistochemically, and synthesized type II collagen. These results show that gelatin/chondroitin sulfate/hyaluronan tri-copolymer has potential for use as a cartilage tissue engineering scaffold. PMID- 14530083 TI - A removal torque of the laser-treated titanium implants in rabbit tibia. AB - The purpose of the present study is to evaluate the significance of different surface textures by comparison of the removal forces for laser-treated and machined titanium screw 8 weeks after the installation in rabbit tibia. A total of 14 screw shaped, commercially pure titanium implants with a length of 5 mm, a diameter of 3.75 mm were grouped as follows: Group A: seven implants left as machined; Group B: seven implants treated with laser method (CSM implant, CSM company, Daegu, Korea) Topographic evaluation was performed with scanning electron microscope (Hitachi S-4200, Japan) to compare the surface structure of laser-treated and machined ones. Installation procedures were done according to Branemark protocol after pre-threading, machined implants were inserted in right tibia metaphysics and laser-treated surface implants were inserted in left ones. Eight weeks post surgically seven rabbits were sacrificed. The implant sites were exposed, and the bone and soft tissues that had formed on top of the implants were carefully removed. Subsequently, the force needed to unscrew the implants (n=14) was measured using a digital torque gauge (Mark-10 corporation, USA). Scanning electron micrographs of the laser-treated and machined control groups demonstrated created a deep and regular honey-comb pattern with small pore, while machined treatment created the typical microscopically grooved and relatively smooth surface characteristic. Eight weeks after implant placement, the average removal torque was 23.58+/-3.71 N cm for the machined implants, 62.57+/-10.44 N cm for the laser-treated implants. The torque measurements yielded statistically significant differences between the machined group and the laser-etched group (p=0.00055) (Wilcoxon's signed-rank test). The laser-treated group achieved higher removal torque values compared to the machined control group. PMID- 14530084 TI - Development of a long-lasting ventricular catheter impregnated with a combination of antibiotics. AB - A ventricular silicone catheter impregnated with a combination of rifampin and a quinolone was developed in order to prevent ventricular shunt related infections. As model substance for the quinolones we used sparfloxacin, because of its specific physicochemical properties resulting in a quantitative detection also in the presence of a second antibiotic. In our study we focused especially on an optimization of the antibiotic release out of the impregnated catheters in order to develop long lasting devices with a broad antimicrobial spectrum. A release optimized catheter was tested with an in vitro colonization test and additionally with a method developed to examine the spread of bacteria on a catheter surface. In vitro experiments showed that the impregnated catheters reduce the colonization with Staphylococcus epidermidis for at least 1 year and prevent the spread of bacteria along the catheter surface. PMID- 14530085 TI - Use of a novel carbon fibre composite material for the femoral stem component of a THR system: in vitro biological assessment. AB - A novel, low elastic modulus femoral component for THR has been developed using a composite of polyetheretherketone and carbon fibre. The objectives of this study were to investigate human osteoblast-like cell and macrophage responses to this material in vitro. Cells were grown on composite discs and controls. Osteoblast attachment and proliferation was not significantly different to that on Ti6Al4V. The levels of alkaline phosphatase activity, Type I collagen production and osteocalcin production were not significantly different to that on Ti6Al4V by the end of the experimental period. Hydrogen peroxide production by macrophages was significantly less than that detected for cells cultured on copper, but was still greater than that detected for cells cultured on tissue culture plastic and Ti6Al4V. Beta-glucoronidase activity was not significantly different to that detected for cells cultured on tissue culture plastic. The in vitro biocompatibility assessment of this composite undertaken in this study showed initial osteoblast attachment at least comparable to that of the tissue culture plastic and Ti6Al4V controls, with proliferation similar to the controls at all time points up to 11 days. Alkaline phosphatase activity was similar to that of Ti6Al4V but reduced compared to tissue culture plastic controls. Whilst hydrogen peroxide production by macrophages was raised on composite surfaces compared to controls, beta-glucoronidase activity and osteoblastic production of Type I collagen and osteocalcin were similar to levels detected on Ti6Al4V. PMID- 14530086 TI - Thermal assembly of a biomimetic mineral/collagen composite. AB - A strategy is described for exploiting temperature driven self-assembly of collagen and thermally triggered liposome mineralization to form a mineralized collagen composite from an injectable precursor fluid. Optical density and rheological experiments demonstrated the formation of a collagen gel when acid soluble type I collagen solutions (1-7 mg/ml) were heated to 24-30 degrees C. Scanning calorimetry experiments demonstrated that mixtures of calcium- and phosphate-loaded liposomes composed of dipalmitoylphosphatidylcholine (90 mol%) and dimyristoylphosphatidylcholine (10 mol%) were stable at room temperature but formed calcium phosphate mineral when heated above 35 degrees C, a consequence of the release of entrapped salts at the lipid chain melting transition. The formation of calcium phosphate mineral induced by triggered release of calcium and phosphate was detected as an endothermic transition (deltaH=6.2+/-1.1 kcal/mol lipid) near the lipid chain melting transition (Tm=37 degrees C). Combining an acid-soluble collagen solution with calcium- and phosphate-loaded liposomes resulted in a liposome/collagen precursor fluid, which when heated from room temperature to 37 degrees C formed a mineralized collagen gel. The dynamic storage modulus of the collagen scaffold increased upon mineralization, and direct nucleation of mineral from the collagen scaffold was detected by electron microscopy. PMID- 14530087 TI - Ion-molecule reactions of gas-phase chromium oxyanions. CrxOyHz-+ O2. AB - Chromium oxyanions, Cr(x)O(y)H(z)(-), were generated in the gas-phase using a quadrupole ion trap secondary ion mass spectrometer (IT-SIMS), where they were reacted with O(2). Only CrO(2)(-) of the Cr(1)O(y)H(z)(-) envelope was observed to react with oxygen, producing primarily CrO(3)(-). The rate constant for the reaction of CrO(2)(-) with O(2) was approximately 38% of the Langevin collision constant at 310 K. CrO(3)(-), CrO(4)(-), and CrO(4)H(-) were unreactive with O(2) in the ion trap. In contrast, Cr(2)O(4)(-) was observed to react with O(2) producing CrO(3)(-) + CrO(3) via oxidative degradation at a rate that was approximately 15% efficient. The presence of background water facilitated the reaction of Cr(2)O(4)(-) + H(2)O to form Cr(2)O(5)H(2)(-); the hydrated product ion Cr(2)O(5)H(2)(-) reacted with O(2) to form Cr(2)O(6)(-) (with concurrent elimination of H(2)O) at a rate that was 6% efficient. Cr(2)O(5)(-) also reacted with O(2) to form Cr(2)O(7)(-) (4% efficient) and Cr(2)O(6)(-) + O (2% efficient); these reactions proceeded in parallel. By comparison, Cr(2)O(6)(-) was unreactive with O(2), and in fact, no further O(2) addition could be observed for any of the Cr(2)O(6)H(z)(-) anions. Generalizing, Cr(x)O(y)H(z)(-) species that have low coordinate, low oxidation state metal centers are susceptible to O(2) oxidation. However, when the metal coordination is >3, or when the formal oxidation state is > or =5, reactivity stops. PMID- 14530088 TI - Rapid and sensitive identification of epitope-containing peptides by direct matrix-assisted laser desorption/ionization tandem mass spectrometry of peptides affinity-bound to antibody beads. AB - A method has been developed for rapid and sensitive identification of epitope containing peptides, based on direct MALDI-MS/MS analysis of epitope-containing peptides affinity bound to affinity beads. This technique provides sequence information of the epitope that allows unambiguous identification of the epitope either by database searching or de novo sequencing. With MALDI-MS, affinity beads with bound peptides can be placed directly on the MALDI target and analyzed. Coupling a MALDI source to an orthogonal injection quadrupole time-of-flight (QqTOF) mass spectrometer allows direct sequencing of the bound peptides. In contrast to ESI-MS/MS, elution of the affinity-bound peptides followed by additional concentration and purification steps is not required, thus reducing the potential for sample loss. Direct mass spectrometric sequencing of affinity bound peptides eliminates the need for chemical or enzymatic sequencing. Other advantages of this direct MALDI-MS/MS analysis of epitope-containing peptides bound to the affinity beads include its sensitivity (femtomole levels) and speed. In addition, direct analysis of peptides on affinity beads does not adversely affect the high mass accuracy of a QqTOF, and database searching can be performed on the MS/MS spectra obtained. In proof-of-principle experiments, this method has been demonstrated on beads containing immobilized antibodies against phosphotyrosine, the c-myc epitope tag, as well as immobilized avidin. Furthermore, de novo sequencing of epitope-containing peptides is demonstrated. The first application of this method was with anti-FLAG-tag affinity beads, where direct MALDI MS/MS was used to determine an unexpected enzymatic cleavage site on a growth factor protein. PMID- 14530089 TI - Intercomparison study on accurate mass measurement of small molecules in mass spectrometry. AB - The results from an intercomparison of accurate mass measurement of a small molecule (molecular weight 475 Da) across a broad range of mass spectrometers are reported. The intercomparison was designed to evaluate the relative capabilities and the optimum methodology of the diverse range of mass spectrometers currently used to record accurate mass measurements. The data will be used as a basis for developing guidance on accurate mass measurement. The need for guidance has resulted from the continued growth in the use of accurate mass measurements for assignment of elemental formula in the chemical and biochemical industries. This has been fuelled by a number of factors and includes the rapid pace of instrument development, which has enabled accurate mass measurements to be made in a less costly, yet robust fashion. The data from the intercomparison will allow us to compare those protocols that produced excellent accuracy and precision with those that produced poorer accuracy and/or precision for each type of mass spectrometer. The key points for best practice will then be established from this comparison for each type of mass spectrometer and accurate mass measurement technique. A compound was sent to the participating laboratories (in the UK, Europe, and USA), the identity of which was not revealed. Each laboratory was asked to record a minimum of five repeat mass measurements of the molecular species using their local protocols and their preferred ionization technique or techniques. To the best of our knowledge there were no interfering (unresolved) ions that originated from the sample. A questionnaire was also completed with the experimental work. The information from the questionnaires was used to evaluate the protocols used to record the measurements. Forty-five laboratories have reported their results. To summarize the performance of mass spectrometers in the intercomparison, magnetic sector field mass spectrometers used in peak matching mode and FTMS reported the highest mean mass measurement accuracy (88 and 83%, respectively, achieved < or =1 ppm). Magnetic sector field mass spectrometers used in voltage scanning produced 60% of the mean mass measurements with accuracy < or =1 ppm. Magnetic sector field mass spectrometers used in magnet scanning modes, quadrupole-TOF and TOF instruments generally achieved mean mass measurement accuracy between 5 and 10 ppm. The two low resolution triple quadrupoles used in the inter-comparison produced mean mass measurement accuracy of <2 ppm. The precision of the data from each instrument and experiment type is an important consideration when evaluating their relative capabilities. Using both the precision and accuracy, it will be possible to define the uncertainty associated with the elemental formulae derived from accurate mass measurements. Therefore, a thorough statistical evaluation of the data is underway and will be presented in a subsequent publication. PMID- 14530090 TI - The use of static pressures of heavy gases within a quadrupole ion trap. AB - The performance of quadrupole ion traps using argon or air as the buffer gas was evaluated and compared to the standard helium only operation. In all cases a pure buffer gas, not mixtures of gases, was investigated. Experiments were performed on a Bruker Esquire ion trap, a Finnigan LCQ, and a Finnigan ITMS for comparison. The heavier gases were found to have some advantages, particularly in the areas of sensitivity and collision-induced dissociation efficiency; however, there is a significant resolution loss due to dissociation and/or scattering of ions. Additionally, the heavier gases were found to affect ion activation and deactivation during MS/MS, influencing the product ion intensities observed. Finally, the specific quadrupole ion trap design and the ion ejection parameters were found to be crucial in the quality of the spectra obtained in the presence of heavy gases. Operation with static pressures of heavy gases can be beneficial under certain design and operating conditions of the quadrupole ion trap. PMID- 14530091 TI - Evaluation of photoinduced changes in stability constants for metal-ion complexes of crowned spirobenzopyran derivatives by electrospray ionization mass spectrometry. AB - The photoinduced changes of metal-ion complexing ability of crowned spirobenzopyran derivatives were studied by using electrospray ionization mass spectrometry (ESI-MS). Stability constants for the complexation with various metal ions in methanol under visible-irradiation conditions were determined for the first time by ESI-MS. It was found that the stability constants of crowned bis(spirobenzopyran) derivatives with metal ions are decreased dramatically by visible irradiation due to the disappearance of powerful ionic interaction between phenolate anion(s) of the merocyanine form of their spirobenzopyran moiety and a metal ion bound to their crown ether moiety, and the decrease in the stability constants is more pronounced for the multivalent metal-ion complexes. A theoretical consideration was also made to attain reliable values of stability complexes for metal-ion complexes of crown compounds. PMID- 14530092 TI - Identification of face-to-face inclusion complex formation of cyclodextrin bearing an azobenzene group by electrospray ionization mass spectrometry. AB - The solution-based self-assembly of native and permethylated cyclodextrins (CD) bearing an azobenzene substituent has been studied by electrospray ionization mass spectrometry (ESI-MS). The results revealed that the CD molecules form either a contact or a face-to-face inclusion complex depending on the interaction of their substituents. The mass spectrometric study further demonstrated that the inclusion complex is formed through the interaction between the host CD cavity and the guest-substituent and that a contact complex is formed by hydrogen bonding of the hydroxyl functions at the rims of the CD molecule. We also found that in order to detect the face-to-face inclusion complex by ESI-MS, the following conditions have to be met: (1) The CD moieties must be permethylated to avoid formation of the contact complex, (2) they must possess a guest-substituent of suitable length, such as an azobenzene moiety, and (3) they must possess an NH(2) or OH group at the substituent terminals for protonation and for detection as cations by ESI-MS. Formation of the inclusion complexes was further confirmed by the synthesis of a capped inclusion dimer and a capped monomer. Collision induced dissociation (CID) experiments have been carried out for the contact, the host-guest inclusion, and the capped inclusion dimers, and the contact complexes are found to be the most stable among them. PMID- 14530093 TI - Efficient determination and evaluation of model cyclodextrin complex binding constants by electrospray mass spectrometry. AB - Electrospray ionization mass spectrometry (ESI-MS) is now routinely used for the detection of cyclodextrin noncovalent complexes, complementing previously established methods. Host-guest complexes formed in solution are also stable for characterization by ESI in the gas phase. This paper reports the first investigations to characterize the stability of three inclusion complexes between beta-cyclodextrin (beta-CD) and three model "guest" molecules, by determining the cyclodextrin compound complex stability constant (K(st)) with the use of mass spectrometric studies. The relative signal intensity of the complexes were monitored in the positive ion mode by mixing each "guest" molecule with an up to 50-fold molar excess of betaCD. A novel linear equation, similar to Benesi Hildebrand, was derived allowing the determination of K(st) for 1:1 stoichiometry in all complexes. These values were compared with the K(st) obtained by spectrophotometric experiments and they were evaluated to be slightly different, indicating the validity of the described method. PMID- 14530094 TI - Mass selective axial ion ejection from a linear quadrupole ion trap. AB - The electric fields responsible for mass-selective axial ejection (MSAE) of ions trapped in a linear quadrupole ion trap have been studied using a combination of analytic theory and computer modeling. Axial ejection occurs as a consequence of the trapped ions' radial motion, which is characterized by extrema that are phase synchronous with the local RF potential. As a result, the net axial electric field experienced by ions in the fringe region, over one RF cycle, is positive. This axial field depends strongly on both the axial and radial ion coordinates. The superposition of a repulsive potential applied to an exit lens with the diminishing quadrupole potential in the fringing region near the end of a quadrupole rod array can give rise to an approximately conical surface on which the net axial force experienced by an ion, averaged over one RF cycle, is zero. This conical surface has been named the cone of reflection because it divides the regions of ion reflection and ion ejection. Once an ion penetrates this surface, it feels a strong net positive axial force and is accelerated toward the exit lens. As a consequence of the strong dependence of the axial field on radial displacement, trapped thermalized ions can be ejected axially from a linear ion trap in a mass-selective way when their radial amplitude is increased through a resonant response to an auxiliary signal. PMID- 14530095 TI - Evaluation of noncovalent interactions between peptides and polyether compounds via energy-variable collisionally activated dissociation. AB - Energy-variable collisionally activated dissociation (CAD) was used to analyze noncovalent interactions of protonated peptide/polyether complexes in a quadrupole ion trap complexes were formed with a series of four polyether host molecules and thirteen peptide molecules. Comparison of dissociation thresholds revealed correlations between the gas-phase basicities of the peptides and polyether molecules and the onset of dissociation. The dissociation thresholds of complexes containing the tripeptides or pentapeptides were inversely proportional to the gas-phase basicities of the sites of protonation of the peptides. Intramolecular hydrogen bonding of the pentapeptides affected the observed dissociation thresholds as well. The dissociation thresholds also scaled proportionally to the gas-phase basicities of the polyethers in the complexes, and the importance of the conformational flexibility of the polyether ligand was confirmed for one of the histidine-containing tripeptide complexes. PMID- 14530096 TI - Fragmentation studies of noncovalent sugar-sugar complexes by infrared atmospheric pressure MALDI. AB - An investigation of sugar-sugar noncovalent complex fragmentation was conducted using a 2.94 microm Er:YAG laser for infrared (IR) atmospheric pressure matrix assisted laser desorption/ionization (AP MALDI) on an ion trap mass spectrometer (ITMS). This approach allowed the analysis of weak noncovalent complexes between a variety of biologically relevant oligosaccharides. The strength of interaction varied with different sugar structures, potentially due to varying strength of hydrogen bonding networks. In some cases, fragmentation of intramolecular sugar bonds preceded breakdown of the noncovalent complex. This result appeared primarily when complexes contained sugars with at least one sialic acid. Globotrios dimers also showed intramolecular fragmentation in preference to breakdown of the noncovalent dimer. This technique will allow further study of sugar-sugar interactions known to play a role in cellular interactions. PMID- 14530097 TI - A chemical ionization study of deuteron transfer initiated propene loss from propoxypyridines. AB - The mechanism of propene loss from the metastable [M + D](+) ions of isomeric 2-, 3-, and 4-n-propoxypyridines and the related isopropoxypyridines has been examined by chemical ionization (CI) and tandem mass spectrometry in combination with deuterium labeling. The [M + D](+) ions were generated with CD(3)OD, CD(3)CN, (CD(3))(2)CO, or pyrrole-D(5) (listed in order of increasing proton affinity) as the CI reagent. The results reveal that the deuteron added in the CI process is not interchanged with the hydrogen atoms of the propyl group prior to propene loss from the metastable [M + D](+) ions of the propoxypyridines. The site selective labeling of the alpha-, beta-, or gamma-position of the propyl group indicates that the [M + D](+) ions of 2-n-propoxypyridine expel propene with formation of an ion-neutral complex composed of a propyl carbenium ion and 2 pyridone. By contrast, the [M + D](+) ions of 3-n-propoxypyridine expel propene by: (1) Formation of ion-neutral complexes, and (2) a conventional 1,5-hydride shift from the beta-position of the n-propyl group to the ring and/or a 1,2 elimination type process. For the 4-isomer, the results suggest the occurrence of propene loss by a 1,2-elimination in addition to the intermediate formation of ion-neutral complexes. Loss of propene with one deuterium atom is the only reaction of the [M + D](+) ions of the isopropoxypyridines labeled at the alpha position of the isopropyl group. The results for the isopropoxypyridines labeled with three deuterium atoms at the beta-position are consistent with: (1) The loss of propene by ion-neutral complex formation and the occurrence of a substantial isotope effect in the subsequent proton/deuteron transfer within the complex, and/or (2) the loss of propene by a 1,2-elimination type reaction. PMID- 14530098 TI - Chemical ionization pathways of polyfluorinated chemicals--a connection to environmental atmospheric processes. AB - A systematic mass spectrometry study of an industrially prolific class of polyfluorinated compounds known as telomers was conducted. The study specifically focused upon polyfluorinated alcohols along with corresponding saturated and alpha,beta-unsaturated fluoroacids. Within each class differing fluoroalkyl chain length homologues were investigated, using negative and positive chemical ionization mass spectrometry (NCI and PCI). In the case of the fluoroalcohols, NCI resulted in the production of more elaborate spectra than the other classes. Moreover, it showed the interesting production of HF(2)(-) and the complex of this species, along with F(-), with the parent molecule. These complexes resulted in the formation of the novel H(2)F(3)(-) ion. Results show that there is significant intra-molecular hydrogen bonding that occurs for these compounds, which influences the molecules fragmentation. This bonding will also influence the fate and disposition through environmental processes (e.g., V(P), k(OH), K(OW), K(OA)) which are affected by molecular geometry. Furthermore, there is an increased accumulation and persistence potential for the molecule as a function of the fluorocarbon chain length. We have shown that in conjunction with the use of mass spectroscopy the engertics of environmental processes for polyfluorinated materials can be established. PMID- 14530099 TI - Elimination of water from the carboxyl group of GlyGlyH+. AB - The elimination of water from the carboxyl group of protonated diglycine has been investigated by density functional theory calculations. The resulting structure is identical to the b(2) ion formed in the mass spectrometric fragmentation of protonated peptides (therefore named "b2" in this study). The most stable geometry of the fragment ion ("b2") is an O-protonated diketopiperazine. However, its formation is kinetically disfavored as it requires a free energy of 58.2 kcal/mol. The experimentally observed N-protonated oxazolone is 3.0 kcal/mol less stable. The lowest energy pathway for the formation of the "b2" ion requires a free energy of 37.5 kcal/mol and involves the proton transfer from the amide oxygen of protonated diglycine to the hydroxyl oxygen. Fragmentation initiated by proton transfer from the terminal nitrogen has also a comparable free energy of activation (39.4 kcal/mol). Proton transfer initiating the fragmentation, from the highly basic terminal nitrogen or amide oxygen to the less basic hydroxyl oxygen is feasible at energies reached in usual mass spectrometric experiments. Amide N-protonated diglycine structures are precursors of mainly y(1) ions rather than "b2" ions. In the lowest energy fragmentation channels, proton transfer to the hydroxylic oxygen, bond breaking and formation of an oxazolone ring occur concertedly but asynchronously. Proton transfer to hydroxyl oxygen and cleavage of the corresponding C-O bond take place at the early stages of the fragmentation step, while ring closure to form an oxazolone geometry occurs at the later stages of the transition. The experimentally observed low kinetic energy release is expected to be due to the existence of a strongly hydrogen bonded protonated oxazolone-water complex in the exit channel. Whereas the threshold energy for "b2" ion formation (37.1 kcal/mol) is lower than for the y(1) ion (38.4 kcal/mol), the former requires a tight transition state with an activation entropy, DeltaS++ = -1.2 cal/mol.K and the latter has a loose transition state with DeltaS++ = +8.8 cal/mol.K. This leads to y(1) being the major fragment ion over a wide energy range. PMID- 14530101 TI - Pectus carinatum: the undertreated chest malformation. PMID- 14530102 TI - Use of the Endo-GIA vascular stapler for hepatic resection. AB - OBJECTIVE: The aim of this study was to analyse the outcome of hepatectomy using the Endo-GIA vascular stapler and to derive a strategy to avoid complications from its use. METHODS: From October 1998 to December 2001, 156 patients underwent major hepatectomy for hepatocellular carcinoma. There were 124 men and 32 women, with a median age of 52.5 years. The Endo-GIA vascular stapler was used for division of the hepatic vein and hepatic duct. Intra- and postoperative clinical data were evaluated. RESULTS: The overall morbidity and mortality rates were 32.1% and 8.3%, respectively. Seventy-six percent of patients did not require blood transfusion. The median blood loss volume was 1 L. There were no complications related to the use of the Endo-GIA vascular stapler in dividing the hepatic veins. One patient (0.6%) had a complication from the use of the stapler for right hepatic duct transection and recovered uneventfully after hepaticojejunostomy. Intraoperative cholangiography was performed in subsequent patients undergoing right trisegmentectomy to avoid biliary complications. CONCLUSIONS: The Endo-GIA vascular stapler is a safe and useful tool to divide the hepatic vein, but surgeons should exercise particular caution to avoid complications when it is used for hepatic duct division. PMID- 14530103 TI - Role of ERCP in the management of predicted mild acute biliary pancreatitis. AB - OBJECTIVE: The role of endoscopic retrograde cholangiopancreatography (ERCP) in mild acute biliary pancreatitis is controversial. This study examined the results of ERCP in patients with predicted mild disease and analysed biochemical and imaging findings in relation to the occurrence of choledocholithiasis. PATIENTS AND METHODS: There were 172 consecutive patients, admitted between January 1998 and December 2000, with the diagnosis of acute pancreatitis. All patients were investigated using transcutaneous ultrasonography and ERCP if biliary aetiology was suspected. Serum bilirubin and alkaline phosphatase were measured, together with abdominal ultrasonographic findings, as potential predictors for choledocholithiasis. RESULTS: Biliary calculus was the aetiology in 62.8% of patients (108/172). Among these 108 patients, 79.6% (86/108) suffered from mild disease. There were only 80 patients who underwent ERCP, and the incidence of choledocholithiasis was 45% (36/80). Although significant correlation was shown between all three measured parameters (bilirubin, alkaline phosphatase and ultrasonographic abnormalities) and choledocholithiasis, their individual sensitivities, specificities and predictive values were low. Nonetheless, if all three tests were normal, the incidence of ductal stones was significantly lower than that in cases with abnormalities in any one of these parameters (4.8% vs 59.3%). CONCLUSION: Routine ERCP is not recommended for patients with mild, acute biliary pancreatitis when there is no biochemical derangement or ultrasonographic evidence of a dilated biliary system. PMID- 14530104 TI - Long-term storage of small surgical instruments in autoclaved packages. AB - BACKGROUND: In most operating theatres, unused sterile instruments must be re sterilized according to preset protocols. Protocols differ among institutions and are not based on strong scientific evidence. OBJECTIVE: To determine and compare the duration of sterility of small instruments packaged in double-layered linen versus plastic-paper envelopes after autoclaving. MATERIALS AND METHODS: Two groups of orthopaedic screws were simultaneously sterilized by autoclaving. In Group 1, each screw was packaged in a double-wrapped linen pack. The screws in Group 2 were individually packaged in an inner wrap of paper and an outer plastic paper envelope that is commercially available. Unwrapped screws in Group 3 served as controls. During the first 48 weeks, five packages were randomly taken from each group, and from 48 weeks to 96 weeks, 20 packages were taken at random and sent for microbial culture. Five screws from Group 3 were also randomly picked with each sample. RESULTS: Up to 96 weeks, no organisms were cultured from any sample from Groups 1 and 2. Almost all samples from Group 3 grew several species of bacteria. CONCLUSION: For small metal instruments, autoclaved packages in double-wrapped linen or double-wrapped plastic-paper combinations can be stored safely for at least 96 weeks. PMID- 14530105 TI - Immunohistochemical distribution of surfactant apoprotein-A in congenital diaphragmatic hernia-II. AB - Morphometric analyses of the immunohistochemical expression of surfactant apoprotein A (SP-A) were carried out on the bronchioles of human congenital diaphragmatic hernia (CDH) neonates and then compared with those in a gestational and postnatal age-matched control group. There was no difference in SP-A expression between lung specimens of the control group and unaffected lungs in the CDH group. However, compared with both these lungs, the ipsilateral lungs of the CDH group showed a significant increase in SP-A expression, namely, the number of SP-A-positive cells per bronchiole, the number of SP-A-positive cells per unit perimeter of bronchiole, and the number of SP-A-positive cells per unit bronchiolar surface area. These results suggest that in lungs of CDH cases, especially ipsilateral lungs, there is a possible delay in the functional maturation or development of SP-A synthesis by the bronchiole, and this retardation may play a role in the postnatal respiratory insufficiency observed in CDH patients. PMID- 14530106 TI - Thoracoscopic surgery for congenital diaphragmatic hernia: a report of nine cases. AB - PURPOSE: To describe the surgical technique and early results of thoracoscopic repair of congenital diaphragmatic hernia (CDH) in children. MATERIALS AND METHODS: A retrospective review was undertaken of patients with CDH who underwent thoracoscopic surgery in our institution over a period of 15 months. There were nine patients, five boys and four girls, ranging in age from 7 days to 8 years. All nine patients underwent surgery under general anaesthesia. Reduction of the hernia contents was carried out using one optical trocar and two operating trocars. Pleural insufflation with carbon dioxide was maintained at a pressure of 2 to 4 mmHg. The hernia defect was repaired using non-absorbable interrupted sutures. RESULTS: The hernia was located on the left side in seven patients and on the right side in two. The mean operative time was 80 minutes. In all patients, the chest tube was removed on the first postoperative day. All patients were discharged on the fifth postoperative day. Chest X-ray and clinical examination 3 months after surgery were normal in all patients. CONCLUSION: The thoracoscopic approach for the repair of CDH in children, including small infants and newborns, is feasible and safe. The technique causes minimal trauma, results in good respiratory function, and promotes early recovery. PMID- 14530107 TI - Clinical and operative findings of choledochal cysts in neonates and infants differ from those in older children. AB - PURPOSE: Significant differences in the clinical and pathological presentation of choledochal cysts between infants and older children have been noted. We developed a new management strategy according to these differences. METHODS: Between 1990 and 2001, we managed 34 cases of type I choledochal cyst. Nineteen patients were more than 1 year old. Fifteen patients underwent surgery before the age of 1 year, and eight before the age of 2 months. One patient was diagnosed prenatally. All patients underwent choledochal cyst excision and hepaticojejunostomy. Symptoms and signs, and findings from physical examination, laboratory tests, and pathology were recorded and analysed. Follow-up periods ranged from 6 months to 11 years. RESULTS: Eight cases who were less than 1 year old suffered from painless jaundice, and choledochal cysts in this group usually ended as a blind pouch. Their cystic amylase and lipase concentrations were within normal limits. Biliary cirrhosis was diagnosed by liver biopsy in three of these patients. In contrast, 17 of the 19 patients who were more than 1 year old had abdominal pain, and 15 of their cysts were connected to the pancreatic duct. Cystic amylase and lipase concentrations were often elevated. All patients had an excellent outcome. CONCLUSION: Diseases with different pathogeneses may cause different clinical manifestations and pathology of choledochal cysts in infants and older children. Some patients develop biliary cirrhosis in the early months of life. Our findings show that it is safe and imperative to treat neonates and infants with choledochal cysts as soon as possible, as delayed surgery may cause severe complications. PMID- 14530108 TI - Renal tubular acidosis secondary to FK506 in living donor liver transplantation: a case report. AB - FK506 is an immunosuppressant that is thought to be less nephrotoxic than cyclosporine A. However, complications due to renal tubular acidosis (RTA) have recently been reported. We report a case of RTA secondary to FK506 administration in liver transplantation. A 6-month-old girl was treated with FK506 after undergoing living donor liver transplantation for fulminant hepatitis. On postoperative day 17, she demonstrated hyperkalaemia and metabolic acidosis; she was diagnosed to have hyperkalaemic distal RTA with aldosterone deficiency (type IV). Intravenous sodium bicarbonate and furosemide, and intrarectal calcium polystyrenesulfonate were administered to correct the acidosis and promote potassium secretion. Thereafter, the FK506 concentration in whole blood gradually decreased, and the hyperkalaemia and metabolic acidosis following RTA improved. RTA is one type of nephrotoxicity induced by FK506, and it is reversible in mild cases when appropriately treated. The mechanism of RTA induced by FK506 has not yet been clearly elucidated. Surgeons and physicians should therefore be aware of the potential for RTA to occur with FK506 after any organ transplantation. The treatment for acidosis and hyperkalaemia should be started as soon as RTA is diagnosed, and the dosage of FK506 should also be reduced if possible. PMID- 14530109 TI - Peutz-Jeghers syndrome in children: high recurrence rate in short-term follow-up. AB - We have managed four cases of Peutz-Jeghers syndrome (PJS) in children. Fathers of three of these patients had PJS. There was also a family history of cancer in three cases, with pancreatic cancer in a father, colonic and laryngeal cancers in a grandfather, and hepatic and gastric cancers in a grandmother. It is presumed that in each of the cases, the largest polyp was responsible for initial symptoms. Preoperative examination revealed additional small polyps in the whole alimentary tract except for the oesophagus. Patients underwent laparotomy to remove the largest polyps and subsequent intraoperative endoscopic polypectomy for other small polyps, to minimize intestinal resection. Follow-up gastrointestinal examinations, including upper gastrointestinal series, small intestinal contrast study, and barium enema, were repeated about once a year. Three of four cases showed recurrent small intestinal polyps, and one required a second laparotomy because of recurrent abdominal pain. In conclusion, patients with PJS occurring in childhood have a strong hereditary family history of cancer and a high incidence of recurrence. Careful follow-up examination is mandatory for the gastrointestinal tract, as well as other solid organs that are susceptible to malignant change, throughout a patient's life. PMID- 14530110 TI - Prenatally diagnosed cystic neuroblastoma: a report of two cases. AB - We report two cases of prenatally diagnosed cystic neuroblastoma (PDCN). In the first case, prenatal ultrasonography (US) at 33 weeks' gestation showed a 30 x 20 mm cyst at the upper pole of the right kidney. The size and content of the mass demonstrated no change during pregnancy. Postnatal US showed no change in the cystic mass 4 weeks after birth compared to the prenatal findings. The infant underwent total resection of the tumour at 28 days of age. In the second case, a left cystic mass measuring 50 x 40 mm was detected in a fetus in the 37th week of pregnancy. Postnatal US showed a cystic mass in the left adrenal gland. The US findings showed no change 18 days after birth and the infant underwent total resection of the tumour at 19 days of age. In both cases, pathological examination revealed a neuroblastoma and all of the biological prognostic factors were favourable. Surgical intervention was necessary for a final histological and biological diagnosis to be made. We recommend that prenatally suspected neuroblastomas should normally undergo surgical intervention, unless tumour size decreases within about 1 month after birth. PMID- 14530111 TI - Huge lymphangioma of the tongue: a case report. AB - Lymphangioma of the tongue is relatively rare and may cause facial structural deformity. Using a combination of a V-shaped and central resection, we successfully treated a 6-year-old girl who had massive lymphangioma of the tongue. Postoperatively, her tongue was located completely within her mouth with good cosmetic results. Sensory and motor nerves to the tongue appeared to be intact. Her speech was also improved. PMID- 14530112 TI - Hepatic foreign body - a sewing needle - in a child. AB - We report a case of a 1-year-old boy with a needle-like foreign body embedded in the liver. The foreign body was incidentally found in the right hypochondrium on routine chest X-ray during a periodic medical examination. He was asymptomatic and there was neither a history of swallowing a needle nor a puncture wound on his body. The results of blood tests and physical examination were entirely within normal limits. Computed tomography scan showed that the needle was completely buried in the liver. At laparotomy, some fibrous tissue and a scar were recognized between the surface of the left lobe of the liver and the parietal peritoneum of the upper abdominal wall. The end of the sewing needle was manually squeezed out and extracted from the liver. From this operative finding, it was assumed that the needle had penetrated the liver through his skin. His postoperative course was uneventful and he was discharged on postoperative day 8. PMID- 14530113 TI - Adrenal cytomegaly: two cases detected by prenatal diagnosis. AB - We report our experience with two cases of adrenal cytomegaly, both of which were detected as cystic adrenal masses during prenatal ultrasonographic examinations. In Case 1, a left suprarenal cystic mass was detected in the fetus at 25 weeks of gestation. The mass, measuring 7 cm in diameter, did not show any change in size and was resected 26 days after birth. In Case 2, a right suprarenal lesion was found at 30 weeks of gestation. The cystic lesion, measuring 2 cm x 1.5 cm, did not change in size and was resected 3 months after birth. Adrenal cytomegaly is still not well known. It is characterized by the presence of large polyhedral cells with eosinophilic granular cytoplasm and enlarged nuclei in the adrenal cortex. This condition is thought to be a degenerative process but not a malignancy. Adrenal cytomegaly rarely forms cysts. It seemed to be impossible to diagnose preoperatively in our cases. Because of the difficulty of differentiating between cystic adrenal cytomegaly and other cystic diseases such as neuroblastoma, operative intervention is required in cases where the cysts do not decrease in size. Further study of a larger number of cases is needed to establish an optimal treatment protocol for these tumours. PMID- 14530115 TI - Prenatal surgery for congenital diaphragmatic hernia. AB - Congenital diaphragmatic hernia (CDH) has a mortality rate of up to 77% despite optimal pre- and postnatal care. Fetuses with liver herniation, a low lung-to head ratio, and an early diagnosis before 24 weeks have a particularly poor prognosis. In utero open repair of these fetuses does not improve patient survival. The PLUG (Plug the Lung Until it Grows) technique was reported to be able to reverse pulmonary hypoplasia in CDH. A foam plug or a titanium clip is used and the trachea can be unplugged using Ex Utero Intrapartum Tracheoplasty (EXIT) at birth. Since hysterotomy causes premature labour, a video-fetoscopic intrauterine technique of tracheal occlusion called Fetendo-PLUG was developed. Compared to those who receive standard postnatal care or fetal tracheal occlusion via open hysterotomy, patients who undergo Fetendo-PLUG are reported to have a higher survival rate of 75% and fewer fetal and maternal complications. A recent refinement is to use a detachable balloon for intratracheal occlusion through a single 5 mm port under real-time ultrasound guidance. Without the need for neck dissection, injury to the recurrent laryngeal nerves and trachea and vocal cord paresis can be minimized. The result of this form of treatment for CDH is promising, but further refinement of fetal instrumentation and development of effective tocolytic drugs are still required. PMID- 14530114 TI - Primary mucinous cystadenoma arising from behind the posterior peritoneum of the descending colon in a child: a case report. AB - This is the first report of a primary mucinous cystadenoma (MCA) arising from behind the posterior peritoneum of the descending colon in a paediatric patient. A large intra-abdominal cystic lesion was found incidentally during renal ultrasonography in a 14-year-old girl. Imaging studies showed a 13 x 9 x 15 cm homogeneous cystic lesion with mild contrast enhancement of the wall. The cyst appeared to originate from the retroperitoneum, but was separated from the left kidney, ovary, and pancreas. At laparotomy, there was a cyst behind the posterior peritoneum of the descending colon. The cyst was successfully excised, and histopathology showed MCA. Although primary MCA in the retroperitoneum is extremely rare in children, it should be considered in the differential diagnosis of an intra-abdominal cyst, since it needs to be excised to eliminate the risk of infection, recurrence, and malignancy. PMID- 14530116 TI - [A decade of child health research in developing countries]. AB - Child mortality continues to be a public health priority world-wide. Even though under-five mortality rates have declined from an estimated 84 per 1000 in 1985-89 to 67 per 1000 in 1999, wide geographical differences persist. During the 1990 World Summit on Children in New York, the world's policy-makers and researchers made improving children's lives by ensuring that each child receives the best care and attention possible their main goal. The strategies identified to achieve this goal have guided and benefited from the progress in child health research throughout the last decade. Indeed, research into the causes of ill-health informs and guides public health programmes to improve the burden of preventable deaths, but it requires sustained and renewed investments for the developing world. In this review paper we have aimed to describe the magnitude of current child morbidity and mortality in the developing world; to review the main advances in child health research since the 1990 World Summit on Children and evaluate the implication of different institutions involved in international child health; to measure the adequacy between priority research setting and public health needs for child health in developing countries; to identify constraints in the application of the research results; and to define key strategies and research challenges that are required in the next decade to respond to child heath needs in developing countries. This assessment is based on a ten-year literature review through Medline and a survey to key international informants in the field of child health in developing countries, using a trilingual open-ended questionnaire. The literature published from January 1990 to June 2001, covering more than 4,700 references, enabled us to review more than 130 articles. We contacted 91 institutions involved in child health, located in 27 countries from five continents, and the response rate to our survey was 49.5%. The current priority health needs of children in developing countries most frequently stated in the literature are the reduction of the burden of malnutrition, diarrhoeal diseases, respiratory diseases, perinatal health, HIV/AIDS, measles and malaria. Based on the survey results, malnutrition (78% of respondents) and perinatal health were the most frequently quoted. Less than 15% of the survey respondents provided an example of a successful application of research in the field of infectious diseases. Political support and conflict of interest are the main constraints mentioned. Malnutrition and non-communicable diseases are considered as main areas of neglect in the field of child health research. Our study shows that the perception of field actors in child health on research priorities does not always correspond to the reality of health needs of children in developing countries. Furthermore, it appears that descriptive epidemiology, evaluation and priority setting strategies are essential tools to guide adequate research activities and further improvement of child health. PMID- 14530117 TI - [Maternal deaths and anesthetics in the Lome (Togo) University Hospital]. AB - AIM: To assess the results of anesthesia practices in a department particularly inadequately staffed with physicians specializing in anesthesiology. SETTING AND METHODS: This six-month prospective study (from January through June 2002) took place in the anesthesiology/intensive care unit of the obstetrics and gynecology department of Lome University Hospital Center. A case report file was completed for each patient, and all anesthetics administered in the obstetrical department (labor and delivery room) were recorded and considered. We examined the perinatal deaths among women who underwent surgery. RESULTS: Anesthetics were administered to 318 women during cesarean delivery (306) and uterine scar repair (12). Nearly all patients (98%) were classified in categories 1 or 2 of the ASA physical status classification system (healthy or mild systemic disease). General anesthesia was induced in 95.9% of the women and spinal bloc used for 4.1%. Thiopental was used most often, and certain drugs, including succinylcholine and ephedrine, were not available as needed. Intraoperative monitoring was essentially nonexistent. Emergency situations accounted for 89.6% of these surgical procedures. Of the 16 cases requiring transfusions, an inadequate supply of blood products or the patient's inability to obtain blood was reported in 14 of the cases. Twelve deaths occurred, for a mortality rate among surgical patients of 3.8%. The principal causes of death were respiratory complications of anesthesia and of pregnancy-related toxemia and the unavailability of hypertonic solutions and blood products. CONCLUSION: The results of this survey show that anesthetics play a role in maternal mortality in Togo. Good practice guidelines adapted to this setting must therefore be developed. PMID- 14530118 TI - [Neonatal prognosis for breech infants delivered vaginally at the Josephine Bongo Maternity Hospital in Libreville, Gabon]. AB - Vaginal delivery of infants in the breech position always presents a risk. OBJECTIVE: To assess the neonatal prognosis of vaginal breech deliveries in an urban maternity hospital in West Africa. EQUIPMENT AND METHOD: This four-year retrospective study, from January 1996 through December 1999, examined all singleton breech deliveries at a gestational age of 32 weeks or later. Each breech delivery was compared with the two singleton births in cephalic presentation that followed. Epi Info 6.04 software was used for data analysis. RESULTS: During the study period, the frequency of breech deliveries in our department was 2.7%, and 83 involved singleton births at or after 32 weeks. The average age of these mothers was 27.9 +/- 5.7 years old (versus 26.4 +/- 5.9) and ranged from 16 to 39 years (versus 16 to 43 years). Average parity was 3.4 +/- 2.2 (versus 3.2 +/- 2.4). Mean uterine height was lower for fetuses in the breech position (30.7 +/- 2.8 cm versus 31.8 +/- 2.6 cm; p=0.04). The frequency of premature deliveries was 33.5% and that of low weight babies (<2,500 g) 35.4%. Both were twice that for babies delivered in the cephalic presentation (17.5% and 17.7%). Neonatal morbidity was 8 times higher for the breech deliveries, with prematurity its main cause. The rate of instantaneous neonatal mortality was 10.8% (versus 2%), that is, 5 times greater. No clinical factors that might account for these deaths was recorded. CONCLUSION: Breech delivery is a risk for newborns. Development of measures and standards for obstetric practice in Gabon would help improve its prognosis. PMID- 14530119 TI - [Pediatry imaging in Abidjan]. AB - The purpose of this study was to inventory the infrastructure and equipment available in Abidjan for paediatric radiology, identify the principal problems of radiological exploration of children and offer suggestions to improve it. MATERIAL AND METHODS: We identified the functional radiological material and the practitioners working in radiological services from 1 July through 31 December 1997. We also examined the records of all paediatric examinations performed during a one-year period in Yopougon University Hospital. RESULTS: The population of Abidjan was estimated (by the National Institute of Statistics) at 3,323,214 in 1998, including 1,208,924 children (36%). There were 31 radiologists, that is, 1 per 101, 526 inhabitants, and 1 X-ray technician for 82,824 inhabitants. The equipment inventory revealed 1 ultrasound scan per 42,531 inhabitants, 1 standard imaging table per 112,403, 1 remote-control table per 242,000, 1 dental radiological system per 1,049,099, and 1 computed tomography system per 1,573,648. Children accounted for 17.4% (1836) of the 10,561 patients examined over one year at the teaching hospital, but there was no specific material for children's examinations. CONCLUSION: Neither human resources nor functional equipment are available in adequate numbers. Radiological equipment appropriate for paediatric exploration must be a priority in the future. . PMID- 14530120 TI - [Geographic changes in exposure to dengue]. AB - Many factors are jointly involved in the changes in the epidemiological and clinical expressions of dengue fever, an arbovirus transmitted by Aedes mosquitoes. In the past forty years, this disease has spread throughout the tropical region, well beyond its original home in Southeast Asia. In some geographic areas, it now appears in a severe haemorrhagic form that requires specialized medical treatment. Many of its mechanisms and features depend on the etiologic agent, human host, and vectors as well as upon bioecologic and socioeconomic factors. Its epidemiological cycles are accordingly quite complex. The threat associated with three current transformations--its geographic extension, epidemiological changes, and aggravation of clinical symptoms- requires an approach that integrates geography and pathology. Such an analysis should provide tools to answer some of the riddles (i.e. specify the epidemiogenic geosystem) by sketching various combinations of factors that reflect the emergence of the disease in specific areas and play a role in its spread from one epidemic area to another. This synthesis about the diverse mutations of the disease calls for a methodical comparison by geographic characteristics and population groups. PMID- 14530121 TI - [A network RER rooted on in vitro readout assays of Plasmodium falciparum sensitivity to chloroquine in the Indian Ocean Region]. AB - Chloroquine has been used as a first line drug to treat uncomplicated malaria cases during the last five decades in Madagascar and in the Comoros Union. The four plasmodial species known to infect humans occur on Madagascar Island. Chloroquine-resistant malaria cases, sometimes only suspected from presumptive malaria cases, have been reported in both countries. Thus, to redefine a strategy and a policy to cure malaria, there is a need to get relevant and updated data. In December 1999, the Madagascan Ministry of Health and the Institut Pasteur de Madagascar formed a network named RER for malaria resistance surveillance. In 2000 and 2001, 18 study sites (17 throughout Madagascar and 1 in Comoros) joined this network. Health-care workers were trained mainly for malaria diagnosis through the use of blood smear examination and for malaria case management. To alleviate the lack of competent medical teams within the health centres, and for technical and logistic reasons, as part of the network activities, it was decided to start with in vitro tests to assess the sensitivity of P.falciparum isolates to chloroquine by means of the isotopic method. Parasitized blood samples were collected from consenting patients. P.falciparum isolates were more predominant (989/1,036). Out of the 564 tests done, 432 (76.6%) could be assessed. Results demonstrated that 94.3% (381/404) of the Madagascan P.falciparum isolates were susceptible to chloroquine. In contrast, chloroquine-resistant isolates were prevalent in Comoros (8/28). The network set-up is presented. The usefulness of the in vivo approach and of the in vitro investigations (chemosusceptibility test and screening of mutations accounting for resistance to chloroquine) to monitor the emergence and the dissemination of chloroquine-resistant parasites is discussed. PMID- 14530122 TI - [Epidemiological features of tumors of the skin and mucosal membranes in the department of dermatology at the Yalgado Ouedraogo National Hospital, Ouagadougou, Burkina Faso]. AB - We conducted a retrospective study of the files of all patients seen from 1 January 1992 through 31 December 1996 with tumors of the skin and mucosal membranes at the Yalgado Ouedraogo National Hospital in order to determine the epidemiologic features of this disease. The records revealed 988 patients presented 1024 tumors, which could be classified into 33 categories. Most of the patients (60.6%) were in the age bracket of 20 to 39 years. Nearly all cases (988 or 96.5%) were benign skin tumors, mainly of infectious origin, especially viral (51.7%). We observed a substantial number of sexually transmissible infections, such as condylomata. We also found 36 cases (3.5%) of malignant tumors, including 29 cases of Kaposi sarcoma, five skin carcinoma (13.8%), three spinocellular and two basocellular; we also noted two borderline malignant tumors: a dermatofibrosar-coma protuberans and a nodular hidradenoma. The elevated prevalence of condyloma (151 cases) may explain the predominance of the 20-39 year age group, which is the most sexually active. Our series also confirmed the relative rarity (3.5%) of cutaneous cancers among African blacks. The predominance of Kaposi sarcoma may be explained by the high prevalence of HIV infection in our country. PMID- 14530123 TI - [Enterobacterial susceptibility to antibiotics in northern Lebanon (1998-2001)]. AB - The purpose of this study was to assess the epidemiology of local enterobacterial susceptibility to antibiotics. Between 1 January, 1998, and 31 December, 2001, we studied the sensitivity of 2,238 Enterobacteria to 26 different antibiotic agents in northern Lebanon, in the Microbiology department and Laboratory of the Islami Hospital, Tripoli, Lebanon. We used the diffusion disk method and complied with the guidelines of the French Microbiology Society antibiogram committee. Urinary samples were the most frequent source (67.5%), followed by blood cultures (12.7%). The dominant species in blood cultures was S. typhi (44.7%). We found 194 strains that produced extended-spectrum beta lactamases (ESBL), with the highest prevalence in Serratia spp. (44.3%), followed by Klebsiella pneumoniae (23.7%), Escherichia coli (20.7%) and Klebsiella oxytoca (11.3%). The global susceptibility of these strains to aminopenicillin was 15%; it reached 30% when combined with clavulanic acid. Susceptibility of the ESBL strains to these agents was 0%. The global susceptibility (and that of the ESBL strains, when greater than 0%) to other antibiotics was as follows: ticarcillin 38.5%, piperacillin 38.5%, piperacillin-tazobactam 88% (64%), imipenem 99.4%, (100%), cefalexin 41%, cefoxitin 65% (40.3%), cefuroxime 75%, amikacin 89%, chloramphenicol 30%, gentamicin 78% (42%), tetracycline 28% (16%), minocycline 30% (18.4%), colistin 67% (75%), nitrofuran 40% (45%), cotrimoxazol 40% (13%), nalidixic acid 53% (5.6%), pefloxacin 63% (23%), ciprofloxacin 71% (39%), and levofloxacin 72% (47%). PMID- 14530124 TI - [Corticotherapy in the otorhinolaryngology service at Brazzaville university hospital]. AB - Corticoids are one of the great therapeutic advances of modern medicine. A retrospective study of corticotherapy conducted in the otorhinolaryngology and cervicofacial surgery service at Brazzaville University Hospital (Centre hospitalier et universitaire de Brazzaville) examined 952 cases treated from 1996 through 2000 and enabled us to determine that: corticoids account for 14% of the treatments prescribed to ENT patients; the patients' average age was 27 years; the most common indications were infections and/or inflammatory disorders (98.2%); local administration was most often prescribed (78.7%, compared with systemic, 21.3%); dexamethasone (60.1%) and betamethasone (19.5%) were prescribed most often; the average duration of treatment was 8 days; adverse reactions were rare (2 cases); treatment was effective in 78% of the cases. PMID- 14530125 TI - [Seroprevalence of the human cytomegalovirus among blood donors and AIDS patients in Bamako]. AB - Our aim was to study the prevalence of the human cytomegalovirus (HCMV) among blood donors and AIDS patients and to examine a clinical correlation between AIDS and HCMV seroprevalence in Bamako (Mali, West Africa). We have used Elisa kits for detecting HIV and HCMV specific antibodies. The HCMV seroprevalence was 89% among AIDS patients, 71% among HIV-infected blood donors and 58% among HIV uninfected blood donors. The HCMV seroprevalence rate was higher among AIDS patients than among HIV-uninfected blood donors (p=0.0000007) and than among HIV infected blood donors (p=0.00146). There was no significant difference between the HCMV seroprevalence among HIV-infected blood donors and among HIV-uninfected blood donors (p=0.0547). There was no relationship between HCMV seroprevalence and sex or age among AIDS patients and blood donors. There was a significant difference on pneumonia among HCMV-infected AIDS patients and among HCMV uninfected AIDS patients (40% versus 0%; p=0.005). Buccal and phaiyngal candidosis, dermatosis, Kaposi's sarcoma and nervous system diseases had the same frequency among HCMV-infected and HCMV-uninfected AIDS patients. PMID- 14530126 TI - [Relevance of reactive fillets in detecting urinary infections in African paediatric environment]. AB - It is quite obvious that African paediatricians are facing numerous challenges. However, their daily fight against frequent pathologies such as malaria, diarrhoeal diseases and respiratory infections should not prevent them from taking into consideration the existence of other pathologies, particularly urinary infections. The clinical picture of urinary infections which is often polymorphic would require a systematic test among infants. Reactive fillets may therefore be relevant in detecting such urinary infections. This survey shows that the best intrinsic values of the fillets used (multistix 8 SG) are reached with leucocytes (sensibility: 95%) and with the combination of leucocytes and nitrites (specificity: 77.1%). The best predictable validities were obtained with the combination of leucocytes and nitrites (positive predictable value: 68.1%) as well as with the combination of leucocytes or nitrites (negative predictable value: 86%). In the daily practice, reactive fillets may turn out to be helpful to the clinician. The negativity of both tests speaks for the isolation and exclusion of urinary infections. On the other hand, the positivity of both tests highlights a strong urinary infection which is subject to a careful and rigorous use of fillets. PMID- 14530127 TI - Type I STS markers are more informative than cytochrome B in phylogenetic reconstruction of the Mustelidae (Mammalia: Carnivora). AB - We compared the utility of five nuclear gene segments amplified with type I sequence-tagged site (STS) primers versus the complete mitochondrial cytochrome b (cyt b) gene in resolving phylogenetic relationships within the Mustelidae, a large and ecomorphologically diverse family of mammalian carnivores. Maximum parsimony and likelihood analyses of separate and combined data sets were used to address questions regarding the levels of homoplasy, incongruence, and information content within and among loci. All loci showed limited resolution in the separate analyses because of either a low amount of informative variation (nuclear genes) or high levels of homoplasy (cyt b). Individually or combined, the nuclear gene sequences had less homoplasy, retained more signal, and were more decisive, even though cyt b contained more potentially informative variation than all the nuclear sequences combined. We obtained a well-resolved and supported phylogeny when the nuclear sequences were combined. Maximum likelihood and Bayesian phylogenetic analyses of the total combined data (nuclear and mitochondrial DNA sequences) were able to better accommodate the high levels of homoplasy in the cyt b data than was an equally weighted maximum parsimony analysis. Furthermore, partition Bremer support analyses of the total combined tree showed that the relative support of the nuclear and mitochondrial genes differed according to whether or not the homoplasy in the cyt b gene was downweighted. Although the cyt b gene contributed phylogenetic signal for most major groupings, the nuclear gene sequences were more effective in reconstructing the deeper nodes of the combined tree in the equally weighted parsimony analysis, as judged by the variable-length bootstrap method. The total combined data supported the monophyly of the Lutrinae (otters), whereas the Melinae (badgers) and Mustelinae (weasels, martens) were both paraphyletic. The American badger, Taxidea taxus (Taxidiinae), was the most basal taxon. Because hundreds of type I STS primer sets spanning the complete genomes of the human and mouse have been published and thus represent many independently segregating loci, the potential utility of these markers for molecular systematics of mammals and other groups is enormous. PMID- 14530128 TI - Estimation of rates-across-sites distributions in phylogenetic substitution models. AB - Previous work has shown that it is often essential to account for the variation in rates at different sites in phylogenetic models in order to avoid phylogenetic artifacts such as long branch attraction. In most current models, the gamma distribution is used for the rates-across-sites distributions and is implemented as an equal-probability discrete gamma. In this article, we introduce discrete distribution estimates with large numbers of equally spaced rate categories allowing us to investigate the appropriateness of the gamma model. With large numbers of rate categories, these discrete estimates are flexible enough to approximate the shape of almost any distribution. Likelihood ratio statistical tests and a nonparametric bootstrap confidence-bound estimation procedure based on the discrete estimates are presented that can be used to test the fit of a parametric family. We applied the methodology to several different protein data sets, and found that although the gamma model often provides a good parametric model for this type of data, rate estimates from an equal-probability discrete gamma model with a small number of categories will tend to underestimate the largest rates. In cases when the gamma model assumption is in doubt, rate estimates coming from the discrete rate distribution estimate with a large number of rate categories provide a robust alternative to gamma estimates. An alternative implementation of the gamma distribution is proposed that, for equal numbers of rate categories, is computationally more efficient during optimization than the standard gamma implementation and can provide more accurate estimates of site rates. PMID- 14530129 TI - Identifying conflicting signal in a multigene analysis reveals a highly resolved tree: the phylogeny of Rodentia (Mammalia). AB - Homoplasy among morphological characters has hindered inference of higher level rodent phylogeny for over 100 years. Initial molecular studies, based primarily on single genes, likewise produced little resolution of the deep relationships among rodent families. Two recent molecular studies (Huchon et al., 2002, Mol. Biol. Evol. 19:1053-1065; Adkins et al., 2003, Mol. Phylogenet. Evol. 26:409 420), using larger samples from the nuclear genome, have produced phylogenies that are generally concordant with each other, but many of the deep superfamilial nodes were still lacking substantial statistical support. Data are presented here for a total of approximately 3,600 base pairs from portions of three different nuclear protein-coding genes, CB1, IRBP, and RAG2, from 19 rodents and three outgroups. Separate analyses, with data partitioned according to both genes and codon position, produced conflicting results. Trees obtained from all partitions of CB1 and RAG2 and those obtained from the first- plus second-position sites of IRBP were generally concordant with each other and the trees from the two recent studies, whereas trees obtained from the third-position sites of IRBP were not. Although the IRBP third-position sites represent only 1/9 of the total data set, combined analyses using either parsimony or likelihood resulted in trees in agreement with the IRBP third-position sites and in disagreement with the remaining 8/9 of the sites from this data set and the two recent multigene studies. In contrast, maximum-likelihood analysis using a site-specific rates model did recover a tree that is highly congruent with the trees in the two recent studies. If the IRBP third-position sites are removed from the current data set, then combined likelihood analyses obtain a tree that is highly congruent with those of the two recent studies. This analysis also provides, for the first time in a study of rodent phylogeny, robust statistical support for every bipartition, with just one exception. This tree divides rodents into two major clades. The first contains Myodonta (Muroidea plus Dipodidae) and the only unresolved trichotomy, from which descend Geomyoidea, Pedetidae, and Castoridae. On the other side of the root is a clade containing Sciuroidea plus Gliridae, and Hystricognathi. Some uncertainty remains on the placement of the root. Trees on which the Hystricognathi are the basal sister group to Myodonta, Geomyoidea, Pedetidae, and Castoridae are also found within a Bayesian 95% credible set, as estimated by Metropolis-coupled Markov chain Monte Carlo sampling. PMID- 14530130 TI - Phylogenetic relationships among calyptraeid gastropods and their implications for the biogeography of marine speciation. AB - Although calyptraeid gastropods are not well understood taxonomically, in part because their simple plastic shells are the primary taxonomic character, they provide an ideal system to examine questions about evolution in the marine environment. I conducted a phylogenetic analysis of calyptraeid gastropods using DNA sequence data from mitochondrial cytochrome oxidase I (COI) and 16S genes and the nuclear 28S gene. The resultant phylogeny was used to examine the biogeographic patterns of speciation in the Calyptraeidae. Parsimony and Bayesian analyses of the combined data sets for 94 calyptraeid operational taxonomic units and 24 outgroups produced well-resolved phylogenies. Both approaches resulted in identical sister-species relationships, and the few differences in deeper topology did not affect biogeographic inferences. The geographic distribution of the species included here demonstrate numerous dispersal events both between the Pacific and Atlantic oceans and across the equator. When parsimony is used to reconstruct the movement from the Pacific to the Atlantic oceans on the phylogeny, there are 12 transitions between oceans, primarily from the Pacific to the Atlantic. When the latitude is coded as north versus south of the equator, the most-parsimonious reconstruction gives the origin of calyptraeids in the north followed by 15 dispersal events to regions south of the equator and no returns to the north. Many clades of the most closely related species are either sympatric or occur along a single coastline. Closely related species can, however, occur in such divergent regions as Southern California and South Africa. There is little evidence for sister-species pairs or larger clades having been split by the Isthmus of Panama or the Benguela upwelling, but the East Pacific Barrier appears to separate the most basal taxa from the rest of the family. PMID- 14530131 TI - Frequent inconsistency of parsimony under a simple model of cladogenesis. AB - Although the conditions under which the parsimony method becomes inconsistent have been studied for almost two decades, the probability that the parsimony method would encounter conditions causing inconsistency under simple models of cladogenesis is unknown. Here, we examine the statistical behavior of the parsimony method under a birth-death model of cladogenesis, when the molecular clock holds. The parsimony method can become inconsistent a high proportion of the time even under this simple model of cladogenesis. When taxon sampling is poor or rates of evolution are high, the probability that parsimony will become inconsistent increases. PMID- 14530132 TI - Hierarchical phylogenetic models for analyzing multipartite sequence data. AB - Debate exists over how to incorporate information from multipartite sequence data in phylogenetic analyses. Strict combined-data approaches argue for concatenation of all partitions and estimation of one evolutionary history, maximizing the explanatory power of the data. Consensus/independence approaches endorse a two step procedure where partitions are analyzed independently and then a consensus is determined from the multiple results. Mixtures across the model space of a strict combined-data approach and a priori independent parameters are popular methods to integrate these methods. We propose an alternative middle ground by constructing a Bayesian hierarchical phylogenetic model. Our hierarchical framework enables researchers to pool information across data partitions to improve estimate precision in individual partitions while permitting estimation and testing of tendencies in across-partition quantities. Such across-partition quantities include the distribution from which individual topologies relating the sequences within a partition are drawn. We propose standard hierarchical priors on continuous evolutionary parameters across partitions, while the structure on topologies varies depending on the research problem. We illustrate our model with three examples. We first explore the evolutionary history of the guinea pig (Cavia porcellus) using alignments of 13 mitochondrial genes. The hierarchical model returns substantially more precise continuous parameter estimates than an independent parameter approach without losing the salient features of the data. Second, we analyze the frequency of horizontal gene transfer using 50 prokaryotic genes. We assume an unknown species-level topology and allow individual gene topologies to differ from this with a small estimable probability. Simultaneously inferring the species and individual gene topologies returns a transfer frequency of 17%. We also examine HIV sequences longitudinally sampled from HIV+ patients. We ask whether posttreatment development of CCR5 coreceptor virus represents concerted evolution from middisease CXCR4 virus or reemergence of initial infecting CCR5 virus. The hierarchical model pools partitions from multiple unrelated patients by assuming that the topology for each patient is drawn from a multinomial distribution with unknown probabilities. Preliminary results suggest evolution and not reemergence. PMID- 14530133 TI - Reliability of Bayesian posterior probabilities and bootstrap frequencies in phylogenetics. AB - Many empirical studies have revealed considerable differences between nonparametric bootstrapping and Bayesian posterior probabilities in terms of the support values for branches, despite claimed predictions about their approximate equivalence. We investigated this problem by simulating data, which were then analyzed by maximum likelihood bootstrapping and Bayesian phylogenetic analysis using identical models and reoptimization of parameter values. We show that Bayesian posterior probabilities are significantly higher than corresponding nonparametric bootstrap frequencies for true clades, but also that erroneous conclusions will be made more often. These errors are strongly accentuated when the models used for analyses are underparameterized. When data are analyzed under the correct model, nonparametric bootstrapping is conservative. Bayesian posterior probabilities are also conservative in this respect, but less so. PMID- 14530134 TI - Performance-based selection of likelihood models for phylogeny estimation. AB - Phylogenetic estimation has largely come to rely on explicitly model-based methods. This approach requires that a model be chosen and that that choice be justified. To date, justification has largely been accomplished through use of likelihood-ratio tests (LRTs) to assess the relative fit of a nested series of reversible models. While this approach certainly represents an important advance over arbitrary model selection, the best fit of a series of models may not always provide the most reliable phylogenetic estimates for finite real data sets, where all available models are surely incorrect. Here, we develop a novel approach to model selection, which is based on the Bayesian information criterion, but incorporates relative branch-length error as a performance measure in a decision theory (DT) framework. This DT method includes a penalty for overfitting, is applicable prior to running extensive analyses, and simultaneously compares all models being considered and thus does not rely on a series of pairwise comparisons of models to traverse model space. We evaluate this method by examining four real data sets and by using those data sets to define simulation conditions. In the real data sets, the DT method selects the same or simpler models than conventional LRTs. In order to lend generality to the simulations, codon-based models (with parameters estimated from the real data sets) were used to generate simulated data sets, which are therefore more complex than any of the models we evaluate. On average, the DT method selects models that are simpler than those chosen by conventional LRTs. Nevertheless, these simpler models provide estimates of branch lengths that are more accurate both in terms of relative error and absolute error than those derived using the more complex (yet still wrong) models chosen by conventional LRTs. This method is available in a program called DT-ModSel. PMID- 14530135 TI - Canonical phylogenetic ordination. AB - A phylogenetic comparative method is proposed for estimating historical effects on comparative data using the partitions that compose a cladogram, i.e., its monophyletic groups. Two basic matrices, Y and X, are defined in the context of an ordinary linear model. Y contains the comparative data measured over t taxa. X consists of an initial tree matrix that contains all the xj monophyletic groups (each coded separately as a binary indicator variable) of the phylogenetic tree available for those taxa. The method seeks to define the subset of groups, i.e., a reduced tree matrix, that best explains the patterns in Y. This definition is accomplished via regression or canonical ordination (depending on the dimensionality of Y) coupled with Monte Carlo permutations. It is argued here that unrestricted permutations (i.e., under an equiprobable model) are valid for testing this specific kind of groupwise hypothesis. Phylogeny is either partialled out or, more properly, incorporated into the analysis in the form of component variation. Direct extensions allow for testing ecomorphological data controlled by phylogeny in a variation partitioning approach. Currently available statistical techniques make this method applicable under most univariate/multivariate models and metrics; two-way phylogenetic effects can be estimated as well. The simplest case (univariate Y), tested with simulations, yielded acceptable type I error rates. Applications presented include examples from evolutionary ethology, ecology, and ecomorphology. Results showed that the new technique detected previously overlooked variation clearly associated with phylogeny and that many phylogenetic effects on comparative data may occur at particular groups rather than across the entire tree. PMID- 14530136 TI - A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. AB - The increase in the number of large data sets and the complexity of current probabilistic sequence evolution models necessitates fast and reliable phylogeny reconstruction methods. We describe a new approach, based on the maximum- likelihood principle, which clearly satisfies these requirements. The core of this method is a simple hill-climbing algorithm that adjusts tree topology and branch lengths simultaneously. This algorithm starts from an initial tree built by a fast distance-based method and modifies this tree to improve its likelihood at each iteration. Due to this simultaneous adjustment of the topology and branch lengths, only a few iterations are sufficient to reach an optimum. We used extensive and realistic computer simulations to show that the topological accuracy of this new method is at least as high as that of the existing maximum likelihood programs and much higher than the performance of distance-based and parsimony approaches. The reduction of computing time is dramatic in comparison with other maximum-likelihood packages, while the likelihood maximization ability tends to be higher. For example, only 12 min were required on a standard personal computer to analyze a data set consisting of 500 rbcL sequences with 1,428 base pairs from plant plastids, thus reaching a speed of the same order as some popular distance-based and parsimony algorithms. This new method is implemented in the PHYML program, which is freely available on our web page: http://www.lirmm.fr/w3ifa/MAAS/. PMID- 14530137 TI - Comparison of likelihood and Bayesian methods for estimating divergence times using multiple gene Loci and calibration points, with application to a radiation of cute-looking mouse lemur species. AB - Divergence time and substitution rate are seriously confounded in phylogenetic analysis, making it difficult to estimate divergence times when the molecular clock (rate constancy among lineages) is violated. This problem can be alleviated to some extent by analyzing multiple gene loci simultaneously and by using multiple calibration points. While different genes may have different patterns of evolutionary rate change, they share the same divergence times. Indeed, the fact that each gene may violate the molecular clock differently leads to the advantage of simultaneous analysis of multiple loci. Multiple calibration points provide the means for characterizing the local evolutionary rates on the phylogeny. In this paper, we extend previous likelihood models of local molecular clock for estimating species divergence times to accommodate multiple calibration points and multiple genes. Heterogeneity among different genes in evolutionary rate and in substitution process is accounted for by the models. We apply the likelihood models to analyze two mitochondrial protein-coding genes, cytochrome oxidase II and cytochrome b, to estimate divergence times of Malagasy mouse lemurs and related outgroups. The likelihood method is compared with the Bayes method of Thorne et al. (1998, Mol. Biol. Evol. 15:1647-1657), which uses a probabilistic model to describe the change in evolutionary rate over time and uses the Markov chain Monte Carlo procedure to derive the posterior distribution of rates and times. Our likelihood implementation has the drawbacks of failing to accommodate uncertainties in fossil calibrations and of requiring the researcher to classify branches on the tree into different rate groups. Both problems are avoided in the Bayes method. Despite the differences in the two methods, however, data partitions and model assumptions had the greatest impact on date estimation. The three codon positions have very different substitution rates and evolutionary dynamics, and assumptions in the substitution model affect date estimation in both likelihood and Bayes analyses. The results demonstrate that the separate analysis is unreliable, with dates variable among codon positions and between methods, and that the combined analysis is much more reliable. When the three codon positions were analyzed simultaneously under the most realistic models using all available calibration information, the two methods produced similar results. The divergence of the mouse lemurs is dated to be around 7-10 million years ago, indicating a surprisingly early species radiation for such a morphologically uniform group of primates. PMID- 14530138 TI - Biotic element analysis in biogeography. AB - Biotic element analysis is an alternative to the areas-of-endemism approach for recognizing the presence or absence of vicariance events in a given region. If an ancestral biota was fragmented by vicariance events, biotic elements or clusters of distribution areas should emerge. We propose a statistical test for clustering of distribution areas based on a Monte Carlo simulation with a null model that considers the spatial autocorrelation in the data. The hypothesis tested is that the observed degree of clustering of ranges can be explained by the range size distribution, the varying number of taxa per cell, and the spatial autocorrelation of the occurrences of a taxon alone. A method for the delimitation of biotic elements which uses model-based Gaussian clustering is introduced. We demonstrate our methods and show the importance of grid size by means of a case study, an analysis of the distribution patterns of southern African species of the weevil genus Scobius. The example highlights the difficulties in delimiting areas of endemism if dispersal has occurred and illustrates the advantages of the biotic element approach. PMID- 14530139 TI - Supertrees are a necessary not-so-evil: a comment on Gatesy et al. PMID- 14530142 TI - Making sense of risk. PMID- 14530144 TI - Science communication and the Swedish acrylamide "alarm". AB - On April 24, 2002 the Swedish National Food Administration along with a group of researchers at the University of Stockholm raised an alarm regarding potential health risks associated with eating fried and baked foods such as potatoes and bread. Scientists had found high levels of acrylamide (up to 500 times more acrylamide than that allowed in drinking water by the World Health Organisation), a substance widely believed to cause cancer, in cooked high starch foods. The outcomes of this "alarm" were immediate. In Sweden sales of chips fell by 30-50 percent over a 3-day period following the press conference, and share prices among several fried food manufacturers fell substantially, as stock analysts were fearful that consumption of fried foods would decrease significantly. Four days after the press conference, however, consumers began eating fried food as normal and a number of researchers and journalists in Sweden and elsewhere took the view that the alarm had been both exaggerated and ill placed. In this study, I evaluate the science communication process associated with the scare, based on a content analysis of a select group of Swedish broad sheets from just previous to the April 2002 press conference to the present time (December 2002). In addition, the study is based on interviews with the various Swedish regulators involved in the process itself (in particular at the Swedish National Food Administration) as well as with the scientists responsible for the study at Stockholm University and relevant journalists and politicians. PMID- 14530145 TI - Going public on acrylamide. PMID- 14530146 TI - Acrylamide: lessons for risk management and communication. PMID- 14530147 TI - Messages influencing college women's tanning bed use: statistical versus narrative evidence format and a self-assessment to increase perceived susceptibility. AB - Understanding the effect of messages and other influences on health decision making has the potential to decrease risky behavior such as tanning bed use. This study explores the effect of type of evidence, self-assessments of risk for skin cancer, and personality factors on intention to use and use of tanning beds among Caucasian female college students. Specifically, it targeted the perceived susceptibility component of the Health Belief Model and its impact on intention to tan as well as changes in actual tanning behavior. College students (N=141) in the southeast United States read randomly assigned messages and self-assessments, filled out surveys, and were later contacted for a follow-up telephone survey. The statistical message was rated higher on information value and also resulted in decreased intention to tan, decreased tanning behavior, and increased perceived susceptibility to skin cancer. The narrative message, in contrast, increased perceptions of realism and also worked to decrease intentions to tan. Additionally, the self-assessment manipulation resulted in increased susceptibility and decreased intention to tan and post tanning behavior. Personality factors explained small portions of variance. Key limitations and directions for future research are also addressed. PMID- 14530148 TI - The influence of peer versus adult communication on AIDS-protective behaviors among Ghanaian youth. AB - Who most influences youth's AIDS-protective behaviors: peers, adults, or a combination of both? This paper explores interpersonal communication about reproductive health information among Ghanaian youth, and the association of this communication with different types of reported AIDS-preventive behaviors. Contacts of peer educators in Ghana were surveyed at three sites during April 1998. Respondents age 11 to 26 years were included in this analysis (N=490). Youth who talked with both peers and adults (n=90) were 2.08 times more likely (95% CI: 1.23, 3.51) to report having done anything to protect themselves from AIDS than those who talked to no one (n=202), while those who talked with peers only (n=150) were 1.71 times more likely (95% CI: 1.10, 2.64) to have done something to protect themselves from AIDS. Youth who talked with adults only were not significantly more likely to protect themselves from AIDS than those who spoke with no one (n=42). Sexually active youth were more than twice as likely to talk to peers as adults. Specific AIDS-protective behaviors reported by youth differed substantially depending on whether their contact source was peers or adults. Understanding the interelationship between peer and adult influence allows program managers to design increasingly effective programs. PMID- 14530149 TI - Chronic disease coverage in Canadian aboriginal newspapers. AB - PURPOSE: To determine the volume and focus of articles on four chronic diseases in newspapers targeting First Nations, Metis, and Inuit in Canada. METHODS: From a sampling frame of 31 Aboriginal newspapers published in English from 1996-2000, 14 newspapers were randomly selected allowing for national and regional representation. Newspaper archives were searched at the National Library of Canada and articles selected if the disease terms cancer, cardiovascular disease, diabetes, or HIV/AIDS appeared in the headline, or in the first or last paragraph of the article. Articles were coded for inclusion of mobilizing information (local, distant, unrestricted, not specified, none) and content focus (scientific, human interest, commercial, other). Cancer articles were categorized by tumor site specificity. Data were analyzed by frequency, cross tabulations, and chi-square analysis. RESULTS: Of 400 chronic disease articles, there were significantly more articles on HIV/AIDS (167 or 41.8%) and diabetes (135 or 33.8%) and few articles on cancer (56 or 14%) and cardiovascular disease (30 articles or 7.5%) (p<0.001). Slightly more than one third (36.5%) of the articles contained mobilizing information to enable readers to take further health action. Mobilizing information was virtually absent from cardiovascular (7/30 or 23%) and diabetes (29/135 or 21.5%) articles. Site specific cancer coverage differed significantly from chance (p<0.001) with 41% of the articles on breast cancer and no articles on lung or colorectal cancers. INTERPRETATION: Given the burden of tobacco-related cardiovascular disease and cancer in Canadian Aboriginal people, the lack of coverage and limited mobilizing information in ethnic newspapers are a missed opportunity for health promotion. PMID- 14530150 TI - A paid radio advertising campaign to promote parent-child communication about alcohol. AB - This study assessed the impact of a paid radio commercial designed to promote parent-child communication about alcohol use and sponsored by the Bureau of Substance Abuse Services, Massachusetts Department of Public Health. A random digit-dial telephone survey of parents or guardians of children ages 10-17 years was conducted after a four-week advertising flight. Respondents with unassisted recall of the commercial more often disagreed that parent-child discussion is useful only if children have begun to experiment with alcohol, and more often reported having three or more parent-child discussions about alcohol compared to those who did not recall the commercial. Findings suggest the potential benefit of paid media campaigns to encourage parents to talk with their children about alcohol. PMID- 14530152 TI - Farnesyltransferase inhibitors as radiation sensitizers. AB - PURPOSE: The inhibition of activated Ras combined with radiotherapy was identified as a potential method for radiosensitization. MATERIALS AND METHODS: Immunoblotting was used to control for prenylation inhibition of the respective Ras isoforms and for changes in activity of downstream proteins. Clonogenic assays with human and rodent tumour cell lines and transfected cell lines served for the testing of radiosensitivity. Xenograft tumours were treated with farnesyl transferase inhibitors and radiation and assayed for ex vivo plating efficiency, regrowth of tumours and EF5 staining for detection of hypoxia. Concurrent treatment with L-778,123 and radiotherapy was performed in non-small cell lung cancer (NSCLC) and head and neck cancer (HNC) patients. RESULTS: Blocking the prenylation of Ras proteins in cell lines with Ras activated by mutations or receptor signalling resulted in radiation sensitization in in vitro and in vivo. The PI3 kinase downstream pathway was identified as a contributor to Ras-mediated radiation resistance. Additionally, increased oxygenation of xenograft tumours was observed after FTI treatment. Combined treatment in a phase I study was safe and effective in NSCLC and HNC. CONCLUSIONS: Tumour cells with activated Ras were sensitized to radiation. Unravelling the underlying mechanisms promises to lead to even more specific drugs with higher potency and safety. PMID- 14530153 TI - Repopulation during fractionated radiotherapy: much has been learned, even more is open. PMID- 14530154 TI - Proliferation and micromilieu during fractionated irradiation of human FaDu squamous cell carcinoma in nude mice. AB - PURPOSE: Previous functional radiobiological experiments demonstrated a significant acceleration of repopulation after 3 weeks and reoxygenation after 12 days of radiotherapy of FaDu tumours. Owing to the temporal coincidence between repopulation and reoxygenation, it was hypothesized that the improved oxygenation status during fractionated irradiation might be the preceding stimulus for increased proliferation. The study investigated whether these changes in repopulation and re-oxygenation are reflected by histological parameters of proliferation and the tumour micromilieu. MATERIALS AND METHODS: Human FaDu squamous cell carcinomas in nude mice were irradiated with three to 18 fractions of 3 Gy daily or every second day under normal blood flow and clamp hypoxia. At different time points, tumours were excised and stained for Ki67, BrdUrd, epidermal growth factor receptor (EGFR) and markers of the micromilieu (HOECHST 33342, pimonidazole, ER-MP12). RESULTS: On average, Ki67 and BrdUrd labelling indices decreased initially and increased again at later times during the course of fractionated radiotherapy. A similar kinetic pattern was found for the staining intensity of the EGFR. The vascular density in the viable tumour area remained constant during the whole course of irradiation, while the perfused fraction of vessels decreased within the first week of irradiation and returned to baseline values after 2 weeks. There was a corresponding increase in perfusion and a decrease in cellular hypoxia. CONCLUSIONS: The histological results were in surprisingly good agreement with the kinetics of clonogen repopulation and re oxygenation determined previously using functional assays. The results support that the kinetics of repopulation of FaDu squamous cell carcinoma in response to fractionated irradiation are determined not only by intracellular processes, but also by a complex interaction of proliferation parameters with a changing microenvironment. PMID- 14530155 TI - Impact of increased cell loss on the repopulation rate during fractionated irradiation in human FaDu squamous cell carcinoma growing in nude mice. AB - PURPOSE: To determine the impact of increased necrotic cell loss on the repopulation rate of clonogenic cells during fractionated irradiation in human FaDu squamous cell carcinoma in nude mice. MATERIALS AND METHODS: FaDu tumours were transplanted into pre-irradiated subcutaneous tissues. This manoeuvre has previously been shown to result in a clear-cut tumour bed effect, i.e. tumours grow at a slower rate compared with control tumours. This tumour bed effect was caused by an increased necrotic cell loss with a constant cell production rate. After increasing numbers of 3-Gy fractions (time intervals 24 or 48 h), graded top-up doses were given to determine the dose required to control 50% of the tumours (TCD50). All irradiations were given under clamp hypoxia. RESULTS: With increasing numbers of daily fractions, the top-up TCD50 decreased from 37.9 Gy (95% CI: 31; 45) after single dose irradiation to 14.1 Gy (8; 20) after irradiation with 15 fractions in 15 days. Irradiation with 18 daily 3-Gy fractions controlled more than 50% of the tumours without a top-up dose. After irradiation with six fractions every second day, the top-up TCD50 decreased to 26.9 Gy (22; 32). No further decrease of the TCD50 was observed after 12 and 18 irradiations every second day. Assuming a constant increase of TCD50 with time, the calculated doubling time of the clonogenic tumour cells (Tclon) was 7.8 days (4.4; 11.3). The Tclon calculated for FaDu tumours growing in pre-irradiated tissues was significantly longer (p=0.0004) than the Tclon of 5.1 days (3.7; 6.5) determined under the same assumptions in a previous study for FaDu tumours growing in normal subcutaneous tissues. CONCLUSIONS: Increased necrotic cell loss by pre-irradiation of the tumour bed resulted in longer clonogen doubling times during fractionated radiotherapy of human FaDu squamous cell carcinoma. This implies that a decreased necrotic cell loss might be the link between reoxygenation and repopulation demonstrated previously in the same tumour model. PMID- 14530156 TI - Is there a relationship between repopulation and hypoxia/reoxygenation? Results from human carcinoma of the cervix. AB - Long overall treatment times are detrimental for cure by radiotherapy and it has been argued that this may be due to repopulation occurring during the course of treatment. However, attempts to predict treatment outcome in relation to tumour proliferation, using pretreatment measurements of kinetic parameters such as Tpot or labelling index (LI) have not met with great success. One possible reason is that hypoxia/reoxygenation is linked to the growth of the tumour and its ability to repopulate. Data from studies in animal models have provided support for this possibility. We made measurement of tumour hypoxia, reoxygenation during treatment and pretreatment measurements of both Tpot and LI in groups of patients with cervix carcinoma undergoing radical radiation treatment. The data show a relationship between pretreatment pO2 measurements and treatment outcome, but reoxygenation did not show any association with treatment outcome. There was no significant association between pretreatment kinetic parameters and treatment outcome, nor was there any evidence of a relationship between pretreatment kinetic parameters and pO2. In the small group of 28 patients whose tumours underwent measurements of both pretreatment kinetic parameters (Tpot, LI) and reoxygenation, there was no relationship between these two sets of measurements. There was also no evidence that a combination of kinetic and reoxygenation measurements could be predictive of treatment outcome. PMID- 14530157 TI - Time factor and treatment strategies in subclinical disease. AB - Rational biological development of treatment strategies for subclinical metastases has lagged behind such efforts with primary cancers: most adjuvant therapies for subclinical disease have been developed empirically, based on clinical observation. This paper reviews recent studies that point to rapid growth of subclinical disease. The effect of rapid growth of occult metastases and undetectable extensions of primary cancer is to increase the radiation dose necessary for their elimination if treatment duration is extended. This increase may be evident even when changes are made to short courses of treatment, consistent with no lag time between the start of treatment and rapid growth or regrowth of subclinical tumour deposits. This provides a strong rationale for avoiding gaps or delays in adjuvant treatments and suggest that accelerated regimens of radiation or chemoradiation may be advantageous in the treatment of subclinical disease provided that the total dose can be maintained or not greatly reduced from those used conventionally. Conversely, an escalation of total dose with a concomitant increase in overall treatment duration may not result in improved control rates because the rapid growth of small clonogen deposits might counterbalance the effect of the higher dose. PMID- 14530158 TI - Dual radiobiological interpretations of retrospective clinical data: the time factor. AB - Dual interpretations are different radiobiological mechanisms that explain theoretically the same observed results. Radiobiological interpretations of the time factor are most frequently based on changes in total dose that produce a given effect. If this dose is increased by different mechanisms (e.g. increasing overall time and decreasing dose per fraction) at the same time, proposals for altered fractionation schemes based on the choice of one or the other mechanism, in principle, can lead to erroneous predictions of outcome. This is especially the case when the analyses are based on retrospective clinical data, where the influence of patient selection is unknown. Examples of dual interpretations taken from the literature on head and neck, melanoma and prostate cancer are discussed. PMID- 14530159 TI - Clinical implications of mucosal regeneration. PMID- 14530160 TI - Time factor for acute tissue reactions following fractionated irradiation: a balance between repopulation and enhanced radiosensitivity. AB - Experimental data for acute radiation-induced skin reactions are reviewed. These show that for dose fractionation schedules with gaps, repopulation is initiated after a lag period. After this lag period, the isoeffective dose for a given level of skin reaction first increases rapidly, but then slows. The timing of the lag period is related to the total turnover time of the tissue under investigation and, for example, is shorter in rodent skin than in pig or human skin. At the point when accelerated repopulation is initiated, there is a major shortening of the turnover time of the target cell population. At this time, there is evidence, for a short period, for an increase in radiosensitivity of the surviving stem cells in a number of acutely responding normal tissues. This effect is clearly illustrated by the results of experiments using sequential dose fractionation schedules. Prolongation of the schedule from 'short' to schedules that include irradiation over the period when the cell turnover is accelerated is associated with a marked increase in tissue radiosensitivity. Clinically, this is best illustrated by a comparison of the effects of accelerated fractionation schedules, involving multiple fractions/day, with daily fractionation schedules. The increase in radiosensitivity produced by the prolongation of the treatment from 2 to 4-5 weeks was equivalent to > or =1 Gy day(-1). Comparable findings were obtained from animal studies. In the oral mucosa of mice, the initiation of accelerated cell proliferation in surviving cells is associated with the loss of dose sparing by subsequent dose fractionation due to the loss of the capacity to repair sublethal damage. Studies in pig and human skin have indicated that increased radiosensitivity is associated with a loss of cells in the G1 phase of the cell cycle. A collation of these two sets of findings suggests that the repair of sublethal damage takes place over this phase of the cell cycle. One clinical implication of these findings is that the alpha/beta ratio for acute skin reaction changes with the length of the overall treatment time; it is approximately 4.0 Gy for 'short' fractionation schedules that avoid any shortening of the cell cycle time. This increases to 11.2-13.3 Gy for schedules given in 3-4 weeks and to approximately 35 Gy for schedules given in 5-6 weeks. Results for pig skin were in total agreement with those for human skin. PMID- 14530161 TI - Proliferation models in tumours. AB - Cell cycle models can be helpful in understanding the organization and kinetics of cellular proliferation. Until recently, models tended to fall into two classes, those assuming that cells normally reside in a quiescent state and are recruited into and through the cell cycle, and those that attempted to accommodate the heterogeneity seen in human tumours. The proliferation plane model combines these two approaches into a unified model encompassing recruitment and retardation in the cell cycle. It provides a framework to re-evaluate current thinking about the factors and mechanisms that contribute to failure in radiotherapy. PMID- 14530162 TI - Modulation of repopulation processes in oral mucosa: experimental results. AB - PURPOSE: Repopulation processes, i.e. the tissue regeneration responses to radiotherapy with increasing overall treatment time, are the predominant factors defining the radiation tolerance of turnover tissues, such as squamous epithelia of the skin or gastrointestinal tract. The purpose was to assess experimental approaches for the modulation, i.e. stimulation of repopulation, in normal oral mucosa. MATERIALS AND METHODS: Numerous studies have been performed to identify and quantify the efficacy of repopulation processes in oral mucosa in experimental animal models, mainly mouse lip and tongue mucosa, and in some clinical studies. However, only a few investigations focused on the stimulation of these processes, aiming at a reduction in oral mucosal side-effects of radiotherapy. The present review summarizes the biological mechanisms underlying effective repopulation, and delineates experimental approaches for effective stimulation of these processes, eventually resulting in an increase in oral mucosal radiation tolerance. RESULTS: Repopulation in oral mucosa is a complex process dominated by a substantial reorganization of the proliferative structure, including a loss of the stem cell division asymmetry and acceleration of stem cell proliferation, as well as abortive divisions of 'sterilized' cells. Repopulation in mouse oral mucosa is more effective if the initial dose intensity (weekly dose) during the first treatment week(s) is increased. Stem cell production occurs mainly during the treatment weeks, while during treatment breaks, including weekends, differentiating (transit) cells are preferentially produced. Stimulation of superficial cell loss, e.g. by topical administration of mild astringent agents, stimulates mucosal proliferation. This translates into increased radiation tolerance to fractionated irradiation in experimental systems, like mouse tongue mucosa. In clinical studies, a reduction of oral mucosal reactions using the same approach was found during an accelerated radiotherapy regimen, but not during conventionally fractionated protocols. Keratinocyte growth factor has been demonstrated to reduce oral mucosal reactions significantly to single dose and fractionated irradiation. This effect is presumably based on an interaction with repopulation processes. CONCLUSIONS: Repopulation in oral mucosa is a highly complex process, which includes a substantial reorganization of the proliferative structure. In experimental models, its efficacy can be modulated by changes in the fractionation protocol, but more effectively by intervention in the regulation processes, e.g. by stimulation of proliferation through enhancement of cell loss. An alternative promising approach is the administration of growth factors, like keratinocyte growth factor, for effective modulation of oral mucosal repopulation. However, selectivity for the normal tissue, as well as biological mechanisms, must be studied in detail in relevant animal models. PMID- 14530163 TI - Epidermal growth factor receptor and its inhibition in radiotherapy: in vivo findings. AB - Increasing evidence shows that dysregulated epidermal growth factor receptor (EGFR) signalling plays an important part in neoplasia. When over expressed or mutated, EGFR is frequently associated with more aggressive tumour growth, poor patient prognosis and resistance of tumours to cytotoxic agents, including radiation. The present studies with murine carcinomas showed that there is an inverse correlation between the level of EGFR and tumour radiocurability. Likewise, the present clinical study in patients with head and neck cancer shows that EGFR over expression correlates with poorer tumour response to radiotherapy. Adding EGFR to tumour cells in vitro protected cells against the cytotoxic action of radiation, whereas blocking EGFR with anti-EGFR antibodies enhanced cell radiosensitivity. A casual relationship between EGFR and increased cellular resistance to radiation was established by transferring the EGFR gene into low EGFR-expressing radiosensitive tumour cells, which then become radioresistant. Radiation activated EGFR and its downstream signalling pathways in radioresistant but not in radiosensitive tumours, and this effect was associated with increased resistance to radiation, and enhanced repopulation in irradiated tumours. Increasing evidence shows that blockage of EGFR or interference with any of the steps in its signal transduction cascade can counteract negative outcomes of EGFR signalling, which has recently been explored as a therapeutic strategy in cancer treatment. The present findings demonstrate that treatment of human tumour xenografts with C225, an anti-EGFR monoclonal antibody, dramatically enhanced tumour response to radiation. Overall, the findings show that over expression of EGFR may serve as a predictor of tumour treatment outcome by radiotherapy and as a therapeutic target to enhance the efficacy of radiotherapy. PMID- 14530164 TI - Selective inhibition of the epidermal growth factor receptor tyrosine kinase by BIBX1382BS and the improvement of growth delay, but not local control, after fractionated irradiation in human FaDu squamous cell carcinoma in the nude mouse. AB - PURPOSE: To investigate the effect of BIBX1382BS, an inhibitor of the epidermal growth factor receptor tyrosine kinase, on proliferation and clonogenic cell survival of FaDu human squamous cell carcinoma in vitro, and on tumour growth and local tumour control after fractionated irradiation over 6 weeks in nude mice. FaDu human squamous cell carcinoma is epidermal growth factor receptor positive and significant repopulation during fractionated irradiation was demonstrated in previous experiments. MATERIALS AND METHODS: Receptor status, receptor phosphorylation, cell cycle distribution, cell proliferation and clonogenic cell survival after irradiation were assayed with and without BIBX1382BS (5 microM) in vitro. Tumour volume doubling time, BrdUrd and Ki67 labelling indices and apoptosis were investigated in unirradiated tumours growing in NMRI nude mice treated daily with BIBX1382BS (50 mg kg(-1) body weight orally) or carrier. Tumour growth delay and dose-response curves for local tumour control were determined after irradiation with 30 fractions within 6 weeks. RESULTS: BIBX1382BS blocked radiation-induced phosphorylation of the epidermal growth factor receptor and reduced the doubling time of FaDu cells growing in vitro by a factor of 4.9 (p=0.008). Radiosensitivity in vitro remained unchanged after incubation with BIBX1382BS for 3 days and decreased moderately after 6 days (p=0.001). BIBX1382BS significantly reduced the volume doubling time of established FaDu tumours in nude mice by factors of 2.6 when given over 15 days (p<0.001) and 3.7 when applied over 6 weeks (p<0.001). When given simultaneously to fractionated irradiation, growth delay was significantly prolonged by an average of 33 days (p=0.003). Local tumour control was not improved by BIBX1382BS. The radiation doses necessary to control 50% of the tumours locally were 63.6 Gy (95% confidence interval 55; 73) for irradiation alone and 67.8 Gy (60; 77) for the combined treatment (p=0.5). CONCLUSIONS: Despite clear antiproliferative activity in rapidly repopulating FaDu human squamous cell carcinoma and significantly increased tumour growth delay when combined with fractionated irradiation, local tumour control was not improved by BIBX1382BS. The results do not disprove that epidermal growth factor receptor inhibition might enhance the results of radiotherapy. However, the results imply that further preclinical investigations using relevant treatment schedules and appropriate endpoints are necessary to explore the mechanisms of action and efficacy of such combinations. PMID- 14530165 TI - Anti-erbB receptor strategy as a gene therapeutic intervention to improve radiotherapy in malignant human tumours. AB - PURPOSE: We explored and quantified the therapeutic potential of using dominant negative EGFR transduction with replication-incompetent adenovirus (Ad-EGFR-CD533 or Ad-CD533) as a genetic approach for radiosensitization in different carcinoma and malignant glioma cell lines in vitro and in established tumour xenografts in vivo. MATERIAL AND METHODS: The cell lines MDA-MB-231, A-431, U-373 MG, U-87 MG and T47D were used. The ErbB expression profiles were quantified by Western blotting. MAPK immune complex assay measured MAPK activity with or without EGFR CD533 expression after ionizing radiation. Radiosensitization was determined and quantified in vitro by colony-formation assays, in vivo by use of an ex vivo-in vitro colony-formation assay after intratumoral infusion of the adenoviral vectors expressing EGFR-CD533 or the control LacZ. RESULTS: Western blotting demonstrated widely varied expression levels of the ErbB receptors in the tested cell lines. Expression of EGFR-CD533 effectively blocked the radiation-induced activation of MAPK, leading to significant radiosensitization in vitro and in vivo. CONCLUSIONS: The radiation-induced ErbB activation can be effectively modulated by a gene therapeutic approach of over-expressing EGFR-CD533 leading to tumour cell radiosensitization after single and repeated radiation exposures both in vitro and in vivo. PMID- 14530166 TI - Perspectives of experimental research on repopulation during radiotherapy. PMID- 14530167 TI - Repopulation in radiation oncology: perspectives of clinical research. AB - Cellular repopulation is one of the most important biological determinants of the clinical outcome of fractionated radiation therapy. A number of randomized controlled trials of altered dose-fractionation have been conducted in patients with squamous cell carcinoma of the head and neck (HNSCC) and the main biological lessons from these are summarised. Data for other tumour histologies are relatively sparse. Further progress in radiotherapy for HNSCC is unlikely to result from altered fractionation alone, but a number of novel strategies for overcoming or exploiting repopulation are being researched. In the next 5 years, the top priorities for clinical and translational research in this field should be the development of clinically applicable predictive assays, functional imaging as an aid to optimize the dose distribution, optimization of combined modality therapies and novel biological strategies specifically targeting tumour cell proliferation. PMID- 14530169 TI - The proteasome: a novel therapeutic target in haematopoietic malignancy. AB - The proteasome plays a key role in regulating protein degradation in eukaryotic cells. A range of synthetic inhibitors of proteasome activity have been developed which have helped elucidate its role in the cell. These inhibitors have selectively induced apoptosis in malignant cells in vitro suggesting that the proteasome may be a novel therapeutic target. First generation proteasome inhibitors are currently showing promise in phase II/III clinical trials for patients with multiple myeloma. PMID- 14530170 TI - In vitro cytotoxic effects of stabilizing sugars within human intravenous immunoglobulin preparations against the human macrophage THP-1 cell-line. AB - Intravenous immunoglobulin (IVIg) is a safe and effective therapy for the treatment of primary and secondary humoral immune deficiencies and autoimmune disorders. Both minor and more serious side effects may occur following IVIg administration in approximately 1-15% of infusions and stabilizing sugars found in IVIg preparations may contribute some of these. In this report, we aimed to determine the cytotoxic effects of IVIg as compared with four stabilizing sugars (glucose, sucrose, maltose and D-sorbitol) found in IVIg preparations on human monocyte-macrophages. The human THP-1 macrophage cell-line was used as a model to determine the effects of stabilizing sugars and IVIg preparations on cell viability and growth. The sugars differentially affected the viability of THP-1 cells. In experiments using doses of the sugars commonly found in IVIg preparations, cell viability and proliferation was unaffected when compared with doses of IVIg typically administered to patients (5 mg/ml). However, in an LDH release cell lysis assay that measures changes in cell permeability, glucose (50 mg/ml) induced significant release of LDH as compared with complete IVIg (5 mg/ml, p<0.0001). Intranucleosomal DNA fragmentation was not detected at therapeutically relevant doses of IVIg. This suggested that THP-1 cell death was not due to apoptosis. We conclude that osmotic stress mediated by the sugars at high doses promoted THP-1 cell death. We propose that IVIg per se is not cytotoxic to the autonomously growing human THP-1 cell-line but rather, the stabilizing sugars used in the preparations are the cytotoxic factors. This observation was evident when preparations of IVIg were used at high concentrations but not at levels one would associate with clinically relevant doses of IVIg. PMID- 14530171 TI - Circulating blasts following chemotherapy in pediatric patients: implications for complete remission definition in acute leukemia. AB - We assessed the incidence of circulating blasts occurring post-chemotherapy in 1000 consecutive pediatric blood samples. Blasts with myeloid morphology (<1-3%) were present in post-chemotherapy samples in 19 of 294 (6.4%) patients with acute leukemia in remission and in 11 of 361 (3.4%) patients with solid tumors, non Hodgkin or Hodgkin lymphoma, and were associated with ANC>1.5x10(9)/l in 13 of 30 (43%) samples, and platelets >100x10(9)/l in 25 of 30 (83%) samples. Our findings suggest that the absence of circulating blasts may not be a prerequisite for remission in acute myeloid leukemia (AML). PMID- 14530172 TI - Immune reconstitution and early infectious complications following nonmyeloablative hematopoietic stem cell transplantation. AB - Non-myeloablative stem cell transplantation (NMT) has been increasingly used in compromised patients who would otherwise have been unable to undergo allotransplant. There is little understanding of the kinetics of immune reconstitution and its influence on infective complications following NMT. The aim of present study was to evaluate lymphocyte subset reconstitution over the first 12 months post-transplant in 15 adult patients receiving NMT with comparison to that of 30 patients grafted with a conventional hemopoietic stem cell transplantation (HSCT). NMT recipients were conditioned with fludarabine based conditioning regimens. Peripheral blood stem cell (PBSC) was the source of stem cells in 13 NMT recipients and in 24 conventional HSCT recipients. Absolute numbers of helper (CD4+) T cells, naive (CD4+ CD45RA+) and memory (CD4+ CD45RO+) T cells as well as suppressor (CD8+) T cells, CD19+ B cells and NK cells were comparable in the two groups at all time points after transplantation. A median value of 200 CD4+ T cells/microl was achieved at 2 months post-transplant by the NMT and HSCT recipients. The CD4:CD8 ratio remained severely depressed throughout the study period. Almost all CD4+ lymphocytes expressed CD45RO antigen in the both groups of patients B lymphocytes showed low counts throughout the entire study period in both groups. Bacteremia and CMV antigenemia occurred respectively in 13 and 36% of the patients in the NMT group and in 15 and 39% of the patients in the HSCT group. Our preliminary data indicate that patients receiving a NMT have a lymphocyte reconstitution similar to that observed in patients who received a conventional HSCT. The incidence of bacteremia and CMV infection were not significantly different between the groups. Nevertheless, due to the small sample size, these results should be considered suggestive rather than definitive. PMID- 14530173 TI - Characterization of clonogenic progenitors in autologous peripheral blood stem cell grafts: evaluation of a simple in vitro assay suitable for routine clinical use. AB - Autologous peripheral blood stem cell (PBSC) transplantation has a low treatment related morbidity and mortality when using appropriate criteria for patient selection and graft quality evaluation. It will be important to use simple and standardised procedures for evaluation of progenitor cell numbers when considering autografting in patients with malignant or non-malignant disorders and increased risk of prolonged posttransplant cytopenia. We determined the number of clonogenic cells in PBSC autografts after 7 days of in vitro culture, and these results were compared with both the total number of colonies and the numbers of colony subsets in conventional 14 days colony assays (colony-forming unit granulocyte-erythrocyte-macrophage-megakaryocyte, CFU-GEMM; CFU-E, CFU-GM; CFU-megakaryocyte). The total colony number after 7 days of culture correlated significantly with (i) the CD34+ cell number; (ii) the total colony number as well as the numbers of erythroid, nonerythroid and mixed colonies in a conventional assay using 14 days of culture; (iii) the number of megakaryocyte colonies. The total colony number after 7 days of in vitro culture is a simple in vitro parameter that seems to reflect the proliferative capacity of various progenitor subsets in PBSC autografts. This simple analysis may be used in combination with other in vitro techniques (e.g. estimation of stem cell viability and CD34+ cell subset analysis) for pretransplant evaluation of autografts. However, the possible clinical use of this parameter has to be examined in prospective clinical studies. PMID- 14530174 TI - Somatic hypermutation patterns in germinal center B cell malignancies. PMID- 14530175 TI - Cytokines in sickle cell disease. AB - Sickle red cells express adhesion molecules including integrin alpha4beta1, CD36, band 3 protein, sulfated glycolipid, Lutheran protein, phosphatidylserine and integrin-associated protein. The proadhesive sickle cells may bind to endothelial cell P-selectin, E-selectin, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), CD36 and integrins leading to its activation. Monocytes also activate endothelium by releasing proinflammatory cytokines like tumor necrosis factor alpha (TNF-alpha) and interleukin 1beta (IL-1beta). Sickle monocytes also express increased surface CD11b and cytoplasmic cytokines TNFalpha and IL-1beta indicating activated state. Polymorphonuclear leukocytes (PMNs) are also activated with reduced L-selectin expression, enhanced CD64 expression and elevated levels of sL-selectin, sCD16 and elastase resulting in increased adhesiveness to the endothelium. Platelets are also activated and secrete thrombospondin (TSP) and cytokine IL-1. They also form platelet- monocytes aggregates causing endothelial cell P-selectin expression. Endothelial cell activation by these multiple mechanisms leads to a loss of vascular integrity, expression of leukocyte adhesion molecules, change in the surface phenotype from antithrombotic to prothrombotic, excessive cytokine production and upregulation of HLA molecules. Furthermore, contraction of these activated endothelial cells leads to exposure of extracellular matrix proteins, such as TSP, laminin, and fibronectin and their participation in adhesive interactions with bridging molecules from the plasma such as von Willebrand factor (vWf) released from endothelial cells, ultimately culminating in vasoocclusion and local tissue ischemia, the pathognomonic basis of vasoocclusive crisis. PMID- 14530176 TI - Gfi-1 oncoproteins in hematopoiesis. AB - The family of Gfi-1 zinc finger transcriptional repressor oncoproteins consists of Gfi-1 and Gfi-1B. Recent gene targeting experiments and mutational screening in humans have revealed an essential role for Gfi-1 and Gfi-1B in hematopoiesis. Mice lacking Gfi-1 are unexpectedly neutropenic. Neutrophil differentiation is abolished and T lymphocyte differentiation is partially blocked in these mice. Heterozygous germline mutations of Gfi-1 causes severe congenital neutropenia (SCN) in humans. Ela2, whose germline mutation is the major contributor to hereditary neutropenias, is repressed in vivo by Gfi-1. Gfi-1B disruption is embryonic lethal due to a block of erythropoiesis. Gfi-1B is required for both erythroid and megakaryocyte development. The ongoing identification of repressed target genes and interacting transcriptional cofactors is helping to unravel the central contributions of these two hematopoietic factors. PMID- 14530177 TI - Pneumoconiosis, coalmine dust and the PFR. PMID- 14530178 TI - Control banding workshop, 4-5 November 2002, London. PMID- 14530179 TI - European experiences in the development of approaches for the successful control of workplace health risks. AB - In recent years, several approaches have been proposed for the application of control banding concepts to the assessment and management of various workplace health and safety risks. Whilst many of the earlier approaches have originated in the UK, several of the most recent examples have been developed in Europe. The European schemes have attempted to build upon the lessons learned from the earlier control banding schemes and to apply them to new areas of health and safety. This paper analyses the evolution of the earlier approaches and reviews the more recent European developments in the context of continuing regulatory and societal demands for the improved assessment and regulation of workplace chemical risks. PMID- 14530180 TI - Solbase: a databank of solutions for occupational hazards and risks. AB - Several attempts have been made to develop strategies for an effective control of workplace hazards. This paper will focus on the results of a European project called Solbase, which is a databank for solutions to occupational hazards and risks. The Safety Science Group of Delft University of Technology in collaboration with TNO Work and Organisation (formerly NIA-TNO) designed Solbase in a series of projects funded by the Dutch Ministry of Social Affairs and Employment and the European Commission. It consists of the design of and software for a databank with an intelligent navigation system allowing users two principal entry points, which correspond to two basic types of solutions. The first entry point is based on the production process, subdivided into the production principle and production function. This entry point provides the dissemination of solutions within and between branches of industry. The second entry point includes the hazard and its emission and transmission as an access point for more conventional occupational hygiene control measures. With the partners of the consortium, from Spain, Italy, Ireland, Germany, the UK and The Netherlands, 535 new and existing solutions throughout Europe and the world were gathered to test the software and the solutions during a field study. Despite the relatively small number of 'test solutions' used, 54% of the search actions in the field study resulted in a useful and suitable solution which the company could actually put into practice. The companies characterized the software as very user friendly. The reproducibility of the coding system for solutions, the classification tree, was satisfactory. Most coders chose the same keywords from the classification tree to describe a corresponding solution. Solbase is a good searching machine for workplace solutions. Especially, the classification of production processes is an inherent guarantee of an exchange of information across the borders of a specific company or branch of industry. PMID- 14530181 TI - A structured strategy for assessing chemical risks, suitable for small and medium sized enterprises. AB - BACKGROUND: A previous study carried out in 20 Belgian companies, especially small and medium-sized enterprises (SMEs), showed that prevention advisors did not use any structured approaches to assess chemical risk. They used their personal judgement and the information contained in manufacturers' Safety Data Sheets to assess the risk. OBJECTIVE AND METHOD: The purpose of the Regetox network is to provide companies with a global approach for assessing chemical health risks. The structured approach proposed consists of two successive steps of increasing complexity. For the first step, we chose a method developed by the INRS (France), the 'ranking of potential risk', which allows the safety officer or staff member to identify hazards and to set priorities among all the supplied products used in the workplace. For the second step, we applied the COSHH method and EASE model established by the UK Health & Safety Executive for assessing 'chemical risk by reference to occupational exposure limits'. The INRS and COSHH Essentials methods were chosen because they define hazards using R-phrases of the European classification system and assess the exposure by using simple information that is easy to collect in the workplace and toxicological databases. A feasibility study conducted in two enterprises shows the usefulness of this approach. In addition to the intrinsic limitations of the methods, the approach showed some limitations related to the inaccuracy of the manufactured safety data sheets and to the collection of the basic information needed for ranking potential risks. CONCLUSION: The use of the Regetox approach needs training of prevention advisors and a strategy involving employers, staff members and workers in collecting basic information and managing chemical risks. Under these conditions, Regetox seems to be a useful tool for chemical risk assessment in SMEs. PMID- 14530182 TI - Evaluation of the HSE COSHH Essentials exposure predictive model on the basis of BAuA field studies and existing substances exposure data. AB - This paper presents an in-house BAuA study on the evaluation of the COSHH Essentials exposure predictive model. External validation is based on measurement data obtained in BAuA field studies performed in various industries, e.g. printing industry and textile industry. In addition, measurement data and information on industrial hygiene provided by the chemical industry within the framework of the Existing Substances Risk Assessment programme are used. Although the evaluated exposure data cover a wide variety of activities and workplace scenarios, there is still a considerable lack of appropriate exposure data, especially for the more stringent control strategies. It was found that the level of agreement between the measurements for solid substances (powders, dusts) and the predicted ranges is reasonably good. The situation is in part different for liquids. In workplaces where organic solvents are used in litre quantities, exposure levels are within the predicted ranges or are often lower. For small scale uses of liquids (millilitre scale), e.g. in carpenters' workshops, there were indications that the exposure levels can exceed the predicted ranges. However, it must be noted that the database is rather small. PMID- 14530183 TI - Chemical management and control strategies: experiences from the GTZ pilot project on chemical safety in Indonesian small and medium-sized enterprises. AB - In 1998 the Deutsche Gesellschaft fur Technische Zusammenarbeit (GTZ) launched the Convention Project on Chemical Safety in developing countries. The project aims to support developing countries in the implementation of the Rotterdam and Stockholm Conventions, create human resources and institutional capacities and to demonstrate via pilot measures how chemical safety in the partner countries can be improved and sustainably implemented in line with international standards. With this objective the development of a Chemical Management Guide (CM Guide) for small and medium-sized enterprises in developing countries has been initiated. The guide describes a step-by-step approach which is based on identifying 'hot spots' as a first step, and making a chemical inventory as a second step. The third step is the continuous improvement of chemical management. In total, there are six tools that aim to support the chemical management process: basic concepts for risk assessment; description of control approaches; using material safety data sheets (MSDSs); risk phrases for hazardous substances; safety phrases for hazardous substances; symbols used for labelling hazardous substances. In the course of the test-implementation of the CM Guide in Indonesia, it was found that MSDSs were not available in most of the smaller companies. In contrast, medium sized and larger companies do have more MSDSs available. It was also found that the way to engage the minds of company owners and managers is with economic arguments related to the loss, waste and expiry of materials, and quality standards expected from importing countries. PMID- 14530184 TI - Developing COSHH Essentials: dermal exposure, personal protective equipment and first aid. AB - The 'control banding' approach in COSHH Essentials combines the potential for harm with the potential for exposure by inhalation to band measures to control exposure at source, as generic strategies. These are simply adapted to specific tasks and circumstances to produce specific control advice. Where it is not possible or practical to use this control advice, the control bands can suggest adequate respiratory protective equipment using 'protection factors'. Proposals in the paper enable the user to identify the right level of respiratory protective equipment (RPE), and to begin selecting suitable RPE. Selection is made through a formatted questionnaire, enabling the user to give the right facts to the supplier. COSHH Essentials applies mainly to exposure by inhalation. However, skin exposure is very common and uptake via the skin can be an important contributor to body dose. This paper examines the factors concerning skin exposure, and the options for banding the potential for harm to the skin or via the skin. Proposals have then been made for dermal exposure control. Planning for emergencies is an important facet of risk control. Proposals are outlined to band chemical hazards for emergency planning according to a minimum of information, i.e. the danger symbol on a product label. PMID- 14530186 TI - Cardiology patient pages. Aging and diseases of the heart. PMID- 14530187 TI - Images in cardiovascular medicine. Wandering ST-segment elevation. PMID- 14530185 TI - From vulnerable plaque to vulnerable patient: a call for new definitions and risk assessment strategies: Part I. AB - Atherosclerotic cardiovascular disease results in >19 million deaths annually, and coronary heart disease accounts for the majority of this toll. Despite major advances in treatment of coronary heart disease patients, a large number of victims of the disease who are apparently healthy die suddenly without prior symptoms. Available screening and diagnostic methods are insufficient to identify the victims before the event occurs. The recognition of the role of the vulnerable plaque has opened new avenues of opportunity in the field of cardiovascular medicine. This consensus document concludes the following. (1) Rupture-prone plaques are not the only vulnerable plaques. All types of atherosclerotic plaques with high likelihood of thrombotic complications and rapid progression should be considered as vulnerable plaques. We propose a classification for clinical as well as pathological evaluation of vulnerable plaques. (2) Vulnerable plaques are not the only culprit factors for the development of acute coronary syndromes, myocardial infarction, and sudden cardiac death. Vulnerable blood (prone to thrombosis) and vulnerable myocardium (prone to fatal arrhythmia) play an important role in the outcome. Therefore, the term "vulnerable patient" may be more appropriate and is proposed now for the identification of subjects with high likelihood of developing cardiac events in the near future. (3) A quantitative method for cumulative risk assessment of vulnerable patients needs to be developed that may include variables based on plaque, blood, and myocardial vulnerability. In Part I of this consensus document, we cover the new definition of vulnerable plaque and its relationship with vulnerable patients. Part II of this consensus document focuses on vulnerable blood and vulnerable myocardium and provide an outline of overall risk assessment of vulnerable patients. Parts I and II are meant to provide a general consensus and overviews the new field of vulnerable patient. Recently developed assays (eg, C-reactive protein), imaging techniques (eg, CT and MRI), noninvasive electrophysiological tests (for vulnerable myocardium), and emerging catheters (to localize and characterize vulnerable plaque) in combination with future genomic and proteomic techniques will guide us in the search for vulnerable patients. It will also lead to the development and deployment of new therapies and ultimately to reduce the incidence of acute coronary syndromes and sudden cardiac death. We encourage healthcare policy makers to promote translational research for screening and treatment of vulnerable patients. PMID- 14530189 TI - Congress of the European Society of Cardiology, 2003. PMID- 14530190 TI - Association between apolipoprotein E alleles and calcific valvular heart disease. AB - BACKGROUND: Studies on apolipoprotein E (apoE) alleles have reported an increased risk of coronary heart disease in patients with the apoE4 allele. Given the risk factor and histological similarities between coronary and calcific valvular heart disease (aortic stenosis [AS] and mitral annular calcification [MAC]), we postulated that apoE alleles might be associated with the development of these valvular lesions. METHODS AND RESULTS: We evaluated the association between apoE alleles and calcific valvular lesions in 802 patients undergoing transthoracic echocardiography using logistic regression analyses. No difference was noted in genotype distribution (P=0.59) or prevalence of apoE4 between those with or without MAC (30% versus 27%, respectively; P=0.57). Compared with patients without AS, the genotype distribution of patients with AS differed significantly (P=0.03), with increasing prevalences of the apoE 4 allele (27% in those without versus 40% in those with AS; P=0.01). In multivariate analyses adjusting for age, gender, low-density lipoprotein cholesterol levels, and coronary artery disease, increasing age and the apoE4 allele were significant independent predictors of AS (odds ratio, 1.94; 95% confidence interval, 1.01 to 3.71; P=0.046), whereas the apoE4 allele was not predictive of MAC. CONCLUSIONS: These findings support extension of the importance of the apoE4 allele beyond atherosclerosis and Alzheimer's disease to calcific AS. PMID- 14530191 TI - Inflammatory cytokines stimulated C-reactive protein production by human coronary artery smooth muscle cells. AB - BACKGROUND: Serum C-reactive protein (CRP) levels are good predictors of the development of cardiovascular events in apparently healthy men and women. CRP has been believed to be produced exclusively by hepatocytes during the acute-phase response. Several lines of evidence have suggested that atherosclerotic arteries can also produce CRP. However, the cell types that produce CRP locally in the atherosclerotic arterial wall have not been clearly identified. METHODS AND RESULTS: Human coronary artery smooth muscle cells (HCASMCs) and human umbilical vein endothelial cells (HUVECs) were incubated with interleukin-1beta (IL-1beta), IL-6, their combination, tumor necrosis factor-alpha (TNF-alpha), or lipopolysaccharide (LPS) at different concentrations. The supernatants were concentrated and analyzed by a high-sensitivity enzyme-linked immunosorbent assay specific for human CRP. RNA was extracted from the HCASMCs for reverse transcriptase-polymerase chain reaction (RT-PCR) using specific primers for the CRP. Maximal CRP production was observed in HCASMCs after 48 hours of incubation with the combination of 25 ng/mL of IL-1beta and 10 ng/mL of IL-6, whereas incubation with IL-1beta or IL-6 alone only modestly induced CRP. Incubation with TNF-alpha (50 ng/mL) or LPS (1000 EU/mL) resulted in an increase in CRP production comparable to the IL-1beta and IL-6 combination. The induction of CRP in HCASMCs was independently confirmed by RT-PCR comparing the relative CRP mRNA levels. The induction of CRP production by HCASMCs was not reproduced in HUVECs, however. CONCLUSIONS: These results demonstrated that HCASMCs, but not HUVECs, could produce CRP in response to inflammatory cytokines. The locally produced CRP could directly participate in atherogenesis and the development of cardiovascular complications. PMID- 14530192 TI - Circulating pregnancy-associated plasma protein a predicts outcome in patients with acute coronary syndrome but no troponin I elevation. AB - BACKGROUND: Risk stratification in troponin (cTn)-negative acute coronary syndrome (ACS) remains a clinical challenge. We investigated the predictive value of circulating pregnancy-associated plasma protein A (PAPP-A), a novel marker of atherosclerotic plaque activity, in these patients. METHODS AND RESULTS: Two hundred consecutive hospitalized ACS patients were included, of whom 136 (69 men and 67 women; mean+/-SD age, 66+/-16 years) remained cTnI-negative for up to 24 hours. PAPP-A was measured at admission, 6 to 12 hours, and 24 hours. During 6 month follow-up, 26 (19.1%) of the cTnI-negative patients reached a primary end point (cardiovascular death, myocardial infarction, or revascularization). At a cutoff level of 2.9 mIU/L, elevated PAPP-A was an independent predictor of adverse outcome (adjusted risk ratio [RR], 4.6; 95% confidence interval, 1.8 to 11.8; P=0.002). Another independent predictor was admission CRP >2.0 mg/L (RR, 2.6; P=0.03). CONCLUSIONS: Measurement of plasma PAPP-A, a zinc-binding matrix metalloproteinase, is a strong independent predictor of ischemic cardiac events and need of revascularization in patients who present with suspected myocardial infarction but remain troponin negative. PMID- 14530193 TI - Endovascular edge-to-edge mitral valve repair: short-term results in a porcine model. AB - BACKGROUND: The edge-to-edge technique is an accepted method for the surgical repair of a regurgitant mitral valve. This study reports the initial use of an endovascular technology that enables a double-orifice edge-to-edge mitral valve repair without cardiopulmonary bypass in an animal model. METHODS AND RESULTS: Adult pigs (n=14) were anesthetized, and left thoracotomy was performed for epicardial echo imaging. Using femoral vein access, a steerable guide catheter was placed transseptally into the left atrium. An implantable clip designed to grasp and approximate the middle scallops of the anterior and posterior mitral leaflets was introduced through the guide catheter. The clip was opened in the left atrium, advanced through the mitral orifice, and retracted to grasp the leaflet edges. When a functional double-orifice valve was confirmed by echo, the clip was closed to coapt the leaflets and detached from the delivery catheter. Before final clip detachment, echo demonstrated a double orifice in all 14 animals. In 2 studies, the clip released from the anterior mitral leaflet. Retrospective analysis of echo images indicated an incomplete grasp of the anterior leaflet. Immediate postmortem examination revealed that the clip successfully approximated the middle scallops of the anterior and posterior leaflets in all 12 double-orifice studies. CONCLUSIONS: This study demonstrates for the first time that an endovascular system can be successfully used to perform the edge-to-edge repair technique in a nondiseased porcine model. This technique is potentially applicable as a percutaneous catheterization laboratory procedure for the treatment of mitral regurgitation in humans. PMID- 14530194 TI - Feasibility and short-term efficacy of percutaneous mitral annular reduction for the therapy of heart failure-induced mitral regurgitation. AB - BACKGROUND: Mitral regurgitation (MR) frequently accompanies congestive heart failure (CHF) and is associated with poorer prognosis and more significantly impaired symptomatic status. Although surgical mitral valve annuloplasty has the potential to offer benefit, concerns about the combined surgical risk and possible effects on ventricular performance have limited progress. We evaluated the feasibility and short-term efficacy of a novel device placed in the coronary sinus to reduce MR in the setting of CHF. METHODS AND RESULTS: CHF and MR were induced in 9 adult sheep by rapid ventricular pacing for 5 to 8 weeks. A mitral annular constraint device was implanted percutaneously through the right internal jugular vein in the coronary sinus and great cardiac vein to create a short-term stable reduction (24.9+/-2.5%) in the mitral annular septal-lateral dimension as assessed echocardiographically. Right and left heart pressures and cardiac output were determined before and 15 minutes after device implantation. MR extent was examined echocardiographically and expressed as a ratio of left atrial area (MR/LAA). After device placement, MR was substantially reduced from an MR/LAA of 42+6% to 4+/-3% (P<0.01). In association, mean pulmonary wedge pressure was significantly reduced (26+/-3 to 18+/-3 mm Hg; P<0.01) and mean cardiac output significantly increased (3.4+/-0.3 to 4.3+/-0.4 L/min; P=0.01). CONCLUSIONS: In this model of CHF, percutaneous placement of a mitral annular constraint device in the coronary sinus resulted in the short-term elimination or minimization of MR and was accompanied in the short term by favorable hemodynamic effects. PMID- 14530195 TI - Endothelial dysfunction and cardiovascular risk prediction in peripheral arterial disease: additive value of flow-mediated dilation to ankle-brachial pressure index. AB - BACKGROUND: Endothelial dysfunction plays a key role in atherogenesis. We prospectively investigated the impact of noninvasive measurement of endothelial function on cardiovascular risk in peripheral arterial disease (PAD). The study was specially aimed at assessing whether brachial artery flow-mediated dilation (FMD) added to the predictive value of ankle-brachial pressure index (ABPI). METHODS AND RESULTS: Of 131 patients monitored for a mean of 23+/-10 months, 18 had a coronary event, 12 a cerebrovascular event, and 9 a peripheral event. The median FMD was lower in patients with an event than in those without (5.8% versus 7.6%, P<0.05), whereas vasodilation to nitroglycerin was similar in the two groups. The cardiovascular event rate was higher in patients with FMD below the median versus those with FMD above the median (P<0.001 by log-rank test). In a Cox proportion hazard model, independent predictors of events were FMD below the median (P<0.01), ABPI below the median (P<0.01), and previous stroke (P<0.02). Similar results were obtained when peripheral events were excluded from the analysis. Below-median ABPI and FMD combined was more accurate in predicting risk (relative risk [RR] 13.0; 95% CI, 3.0 to 56.2; P<0.01) than ABPI (RR, 6.4; 95% CI, 1.4 to 29.1; P<0.02) and FMD (RR, 4.8; 95% CI, 1.1 to 23.3; P<0.05) alone. CONCLUSIONS: A low brachial artery FMD is an independent predictor of cardiovascular risk in patients with PAD and adds to the prognostic value of ABPI, which is currently the most powerful prognostic indicator in PAD. PMID- 14530196 TI - Association of insulin resistance, hyperleptinemia, and impaired nitric oxide release with in-stent restenosis in patients undergoing coronary stenting. AB - BACKGROUND: Previously undiagnosed diabetes, impaired glucose tolerance, and insulin resistance are common in patients with acute myocardial infarction and coronary heart disease (CHD) and might be involved in early restenosis after stent implantation. To evaluate whether markers of insulin resistance syndrome, including leptin, and endothelial dysfunction are related to increased rate of early restenosis, we studied nondiabetic patients with CHD after successful coronary stenting. METHODS AND RESULTS: Both patients with CHD undergoing coronary stenting (120 patients) and control subjects (58 patients) were submitted to an oral glucose tolerance test (OGTT). Fasting leptin levels and fasting and postglucose load insulin sensitivity were assessed. Endothelial function was measured by nitrite and nitrate release (NOx) during OGTT. More than 50% of patients treated with stent implantation presented impaired glucose tolerance or type 2 diabetes, which was previously undiagnosed. These patients also had higher glucose, insulin, and leptin levels than control subjects. Among the stented patients, insulin and leptin levels were higher in patients with restenosis than in patients without restenosis. A significant increase in NOx levels was found during OGTT both in patients without restenosis and in control subjects. On the contrary, NOx profiles were blunted in patients with restenosis. At multiple regression analysis, only DeltaAUC-NOx areas and insulin sensitivity index showed an independent correlation with the minimal lumen diameter at follow up. CONCLUSIONS: We demonstrated that insulin resistance and endothelial dysfunction are independent predictors of early restenosis after coronary stenting. PMID- 14530197 TI - Sulfatides: targets for anti-phospholipid antibodies. AB - BACKGROUND: Sulfatides are sulfated glycosphingolipids expressed on the surface of erythrocytes, leukocytes, and platelets. Sulfatides interact with several cell adhesion molecules involved in hemostasis. Beta2-glycoprotein I is an anionic phospholipid-binding plasma protein, and the phospholipid-bound form is the target for most anti-phospholipid antibodies that are associated with recurrent thrombosis, miscarriages, and neurological symptoms. In this study, we examined whether beta2-glycoprotein I forms a complex with sulfatides and thereby becomes a target for anti-phospholipid antibodies. METHODS AND RESULTS: Beta2 glycoprotein I binds to surface-bound sulfatides but not to other glycolipids, such as ceramide, cerebrosides, sphingomyelin, or ganglioside. At a sulfatide coating density of 1 microg/well, beta2-glycoprotein I reaches half-maximal binding at 2.5 microg/mL, and the binding is saturated at 10 microg/mL. The binding of beta2-glycoprotein I also depends on the coating density of sulfatides in the well. At a constant beta2-glycoprotein I concentration of 5 microg/mL, maximal binding of beta2-glycoprotein I is observed at a coating density of 1 mug/well. The serum from 14 patients with anti-cardiolipin antibodies, a subset of anti-phospholipid antibodies, bound to sulfatide-bound beta2-glycoprotein I and previous absorption on cardiolipin-coated surfaces decreased the immunoreactivity toward sulfatide-beta2-glycoprotein I complex by >50% in 12 of 14 patients. Furthermore, immunoaffinity-purified anti-cardiolipin antibodies from 4 of 5 patients reacted with sulfatide-bound beta2-glycoprotein I. CONCLUSIONS: These results show that not only anionic phospholipids, as commonly known, but also sulfatides are targets for most anti-phospholipid antibodies. We therefore postulate that interactions of these antibodies with sulfatides may contribute to some of the clinical symptoms of the anti-phospholipid antibody syndrome. PMID- 14530198 TI - Health-related quality of life is better for cardiac arrest survivors who received citizen cardiopulmonary resuscitation. AB - BACKGROUND: This study evaluated the prehospital factors associated with better health-related quality of life for survivors of out-of-hospital cardiac arrest. METHODS AND RESULTS: This prospective, 20-community, cohort study involved consecutive, adult out-of-hospital cardiac arrest patients who survived to 1 year. Patients were contacted by telephone and evaluated for the Health Utilities Index Mark III (HUI3), which describes health as a utility score on a scale from 0 (dead) to 1.0 (perfect health). The 8091 cardiac arrest patients had overall survival rates of 5.2% to hospital discharge and 4.0% to 1 year. We successfully contacted and evaluated 268 of 316 (84.8%) of known 1-year survivors. The median HUI3 score was 0.80 (interquartile range, 0.50 to 0.97), which compares well with age-adjusted values for the general population (0.83). Logistic regression identified 2 factors independently associated with very good quality of life (HUI3 >0.90) and their odds ratios (95% CIs), as follows: age 80 years or older, 0.3 (0.1 to 0.84), and citizen-initiated cardiopulmonary resuscitation (CPR), 2.0 (1.2 to 3.4) (Hosmer-Lemeshow goodness-of-fit statistic, 0.74). CONCLUSIONS: This study is the largest ever conducted for out-of-hospital cardiac arrest survivors, clearly shows that these patients have good quality of life, and is the first to demonstrate that citizen-initiated CPR is strongly and independently associated with better quality of life. These results emphasize the importance of optimizing community citizen CPR readiness. Given the low rate of citizen-initiated CPR in many communities, we believe that local and national initiatives should vigorously promote the practice of bystander CPR. PMID- 14530199 TI - Idiopathic left bundle-branch block-shaped ventricular tachycardia may originate above the pulmonary valve. AB - BACKGROUND: Idiopathic left bundle-branch block (LBBB)-like ventricular tachycardia (VT) is considered to originate in the right ventricular outflow tract (RVOT) or from the aortic root. Both regions are derived from the embryonic outflow tract. We now report that also the pulmonary trunk can give rise to VT, suggesting a common etiology of these tachycardias. METHODS AND RESULTS: We studied 6 patients with symptomatic idiopathic LBBB-VT using electrophysiological mapping techniques. The VT origin was determined by analyzing the electrograms and the angiographic location of the catheter tip at the successful ablation site or the earliest activation site. Eight VTs were induced. Two VTs, with a mean earliest endocardial activation time of -5 and -20 ms and optimal pace mapping, were successfully ablated in the RVOT. In the remaining 6 VTs, the earliest activation site was found in the pulmonary artery, and, at this site, a sharp potential was present -38+/-12 ms before the QRS in 5 VTs. The mean earliest endocardial activation time in the RVOT was -1+/-2 ms. Ablation was attempted in 5 of 6 VTs and resulted in an acutely successful procedure. After a mean follow up of 10+/-4 months, 1 of 5 patients had a recurrence. CONCLUSIONS: The site of origin of idiopathic LBBB-VT can be in the root of the pulmonary artery, suggesting a myocardial connection from this site to the RVOT. If no good criteria for ablation in the RVOT are found, detailed mapping of the pulmonary artery should be performed. PMID- 14530200 TI - Immunoglobulin M type of autoantibodies to oxidized low-density lipoprotein has an inverse relation to carotid artery atherosclerosis. AB - BACKGROUND: Lipoprotein oxidation plays an important part in atherogenesis. Autoantibodies to oxidation-specific epitopes of LDL occur in plasma and atherosclerotic lesions of humans and animals. The potential role of these autoantibodies in atherogenesis still remains unsolved. We studied the relationship between different isotypes of autoantibodies to copper-oxidized LDL and malondialdehyde-modified LDL (MDA-LDL) and carotid artery intima-media thickness (IMT) in a population-based cohort of 1022 middle-aged men and women. In addition, we studied the relation of C-reactive protein (CRP) to IMT. METHODS AND RESULTS: The levels of IgM, IgG, and IgG2 autoantibodies binding to MDA-LDL and copper-oxidized LDL were determined in plasma samples by chemiluminescence based ELISA. IMT and the number of plaques were measured ultrasonographically. The subjects were divided into tertiles for antibody titers. We found an inverse association between IMT and IgM autoantibody titers to MDA-LDL that remained statistically significant after adjusting for age, gender, LDL cholesterol, systolic blood pressure, CRP, and smoking. CRP was not independently associated with IMT. CONCLUSIONS: These results show that IgM autoantibodies to MDA-LDL have an inverse association with carotid atherosclerosis. The possible implications of this finding are discussed. PMID- 14530201 TI - Impact of viability, ischemia, scar tissue, and revascularization on outcome after aborted sudden death. AB - BACKGROUND: Survivors of aborted sudden death attributable to ventricular arrhythmias in the presence of coronary artery disease are at risk for recurrences. The substrate underlying these arrhythmias is not clear, and therefore the relation between ischemia, viability, scar tissue (and revascularization), and the incidence of ventricular arrhythmias (and survival) was studied over up to 3 years. METHODS AND RESULTS: One hundred fifty-three survivors of sudden death underwent stress-rest perfusion imaging. Patients with ischemic/viable myocardium (n=73) were revascularized if possible. Final antiarrhythmic therapy was based on the outcome of electrophysiological testing or left ventricular ejection fraction (LVEF). Implantation of a defibrillator was performed in 112 (72%) patients. During 3-year follow-up, 15 cardiac deaths occurred and 42 (29%) patients had recurrent ventricular arrhythmias. Patients with events (death or recurrence) exhibited more often a severely depressed LVEF (< or =30%), more extensive scar tissue, and less ischemic/viable myocardium on perfusion imaging and less frequently underwent revascularization. Multivariate analysis identified extensive scar tissue and LVEF < or =30% as the only predictors of death/recurrent ventricular arrhythmias. CONCLUSIONS: In patients with aborted sudden death, extensive scar tissue and severely depressed LVEF are the only predictors of death or recurrent ventricular arrhythmias. These patients should be considered for implantation of a defibrillator. PMID- 14530202 TI - Polymorphism of the serotonin transporter gene and pulmonary hypertension in chronic obstructive pulmonary disease. AB - BACKGROUND: The serotonin transporter (5-HTT) is involved in the pulmonary artery smooth muscle hyperplasia that leads to pulmonary hypertension (PH). Because hypoxia and 5-HTT gene polymorphism control 5-HTT expression, we examined 5-HTT gene polymorphism and PH in hypoxemic patients with advanced chronic obstructive pulmonary disease (COPD). METHODS AND RESULTS: In 103 patients with COPD recruited in France (n=67) and the UK (n=36), we determined 5-HTT gene polymorphism and pulmonary artery pressure (PAP) measured during right heart catheterization (France) or Doppler echocardiography (UK). Ninety-eight subjects from the 2 countries served as control subjects. The distribution of 5-HTT gene polymorphism did not differ between patients and control subjects. In patients carrying the LL genotype, which is associated with higher levels of 5-HTT expression in pulmonary artery smooth muscle cells than the LS and SS genotypes, PH was more severe than in LS or SS patients. Mean PAP values in patients from France with the LL, LS, and SS genotypes were 34+/-3, 23+/-1, and 22+/-2 mm Hg (mean+/-SEM), respectively (P<0.01). Corresponding systolic PAP values in the UK were 40+/-3, 28+/-3, and 24+/-3 mm Hg, respectively (P<0.01). Compared with control subjects, platelet 5-HTT protein was increased in COPD patients in proportion to the hypoxemia level, and strong 5-HTT immunostaining was observed in remodeled pulmonary arteries from COPD patients. CONCLUSIONS: 5-HTT gene polymorphism appears to determine the severity of PH in hypoxemic patients with COPD. Because PH is an important prognostic factor in this disease, recognition of patients at risk for PH should be helpful in managing COPD. PMID- 14530203 TI - Physiological genomics of human arteries: quantitative relationship between gene expression and arterial stiffness. AB - BACKGROUND: Previous genomic studies with human tissues have compared differential gene expression between 2 conditions (ie, normal versus diseased) to identify altered gene expression in a binary manner; however, a potentially more informative approach is to correlate the levels of gene expression with quantitative physiological parameters. METHODS AND RESULTS: In this study, we have used this approach to examine genes whose expression correlates with arterial stiffness in human aortic specimens. Our data identify 2 distinct groups of genes, those associated with cell signaling and those associated with the mechanical regulation of vascular structure (cytoskeletal-cell membrane extracellular matrix). Although previous studies have concentrated on the contribution of the latter group toward arterial stiffness, our data suggest that changes in expression of signaling molecules play an equally important role. Alterations in the profiles of signaling molecules could be involved in the regulation of cell cytoskeletal organization, cell-matrix interactions, or the contractile state of the cell. CONCLUSIONS: Although the influence of smooth muscle contraction/relaxation on arterial stiffness could be controversial, our provocative data would suggest that further studies on this subject are indicated. PMID- 14530204 TI - Tumor necrosis factor-alpha inhibits insulin's stimulating effect on glucose uptake and endothelium-dependent vasodilation in humans. AB - BACKGROUND: Inflammatory mechanisms could be involved in the pathogenesis of both insulin resistance and atherosclerosis. Therefore, we aimed at examining whether the proinflammatory cytokine tumor necrosis factor (TNF)-alpha inhibits insulin stimulated glucose uptake and insulin-stimulated endothelial function in humans. METHODS AND RESULTS: Healthy, lean male volunteers were studied. On each study day, 3 acetylcholine (ACh) or sodium nitroprusside (SNP) dose-response studies were performed by infusion into the brachial artery. Before and during the last 2 dose-response studies, insulin and/or TNF-alpha were coinfused. During infusion of insulin alone for 20 minutes, forearm glucose uptake increased by 220+/-44%. This increase was completely inhibited during coinfusion of TNF-alpha (started 10 min before insulin) with a more pronounced inhibition of glucose extraction than of blood flow. Furthermore, TNF-alpha inhibited the ACh forearm blood flow response (P<0.001), and this inhibition was larger during insulin infusion (P=0.01) but not further increased by NG-monomethyl-L-arginine acetate (P=0.2). Insulin potentiated the SNP response less than the ACh response and the effect of TNF-alpha was smaller (P<0.001); TNF-alpha had no effect on the SNP response without insulin infusion. Thus, TNF-alpha inhibition of the combined response to insulin and ACh was likely mediated through inhibition of NO production. CONCLUSIONS: These results support the concept that TNF-alpha could play a role in the development of insulin resistance in humans, both in muscle and in vascular tissue. PMID- 14530205 TI - Noninvasive imaging of transgene expression by use of positron emission tomography in a pig model of myocardial gene transfer. AB - BACKGROUND: Radionuclide imaging of reporter gene expression may be useful for noninvasive monitoring of clinical cardiac gene therapy. Experience until now, however, has been limited to small animals. METHODS AND RESULTS: To evaluate feasibility in a clinically applicable setting, pigs were studied by conventional positron emission tomography (PET) 2 days after regional intramyocardial injection of control adenovirus or adenovirus carrying herpesviral thymidine kinase reporter gene (HSV1-tk). Myocardial blood flow was quantified by use of [13N]ammonia. Subsequently, kinetics of the reporter substrate [124I]-2'-fluoro 2'-deoxy-5-iodo-1-beta-d-arabino-furanosyluracil (FIAU) were assessed over a period of 2 hours. Areas infected with adenovirus expressing HSV1-tk showed significantly elevated FIAU retention during the first 30 minutes after injection. At later times, washout was observed, and retention was not different from that in areas infected with control virus or remote myocardium. Early in vivo FIAU uptake correlated with ex vivo images, autoradiography, and immunohistochemistry for reporter gene product after euthanasia. After intramyocardial injection of both adenoviruses, myocardial blood flow was mildly elevated compared with that in remote areas, consistent with histological signs of regional inflammation. CONCLUSIONS: In vivo quantification of regional myocardial transgene expression is feasible with clinical PET methodology, the radioiodinated reporter probe FIAU, and the HSV1-tk reporter gene. Radioactivity efflux after specific initial uptake was not observed previously in tumor studies, suggesting that tissue-specific differences in nucleoside metabolism influence reporter probe kinetics. By coregistering reporter gene expression with additional biological parameters such as myocardial blood flow, PET allows for noninvasive characterization of the success of cardiac gene transfer along with its functional correlates. PMID- 14530206 TI - Transfer for primary angioplasty versus immediate thrombolysis in acute myocardial infarction: a meta-analysis. AB - BACKGROUND: The benefit of primary percutaneous coronary intervention (PCI) over thrombolysis has been clearly demonstrated in acute myocardial infarction (AMI). However, the best therapeutic strategy for a patient with AMI presenting to acute care services without catheterization facilities remains under debate. Our objective was to gather all available information from clinical trials comparing transfer of patients experiencing AMI for angioplasty versus immediate thrombolysis. METHODS AND RESULTS: We performed a meta-analysis of all data available from published randomized trials and from presentations in scientific sessions of major cardiology congresses comparing the 2 strategies. The primary end point was the combined criteria (CC) of death/reinfarction/stroke as defined in each trial. Relative risk (RR) evaluated the treatment effect. We identified 6 clinical trials including 3750 patients. Transfer time was always <3 hours. The CC was significantly reduced by 42% (95% confidence interval [CI] 29% to 53%, P<0.001) in the group transferred for primary PCI compared with the group receiving on-site thrombolysis. When CC parameters were considered separately, reinfarction was significantly reduced by 68% (95% CI, 34% to 84%; P<0.001) and stroke by 56% (95% CI, -15% to 77%; P=0.015). There was a trend toward reduction in all-cause mortality of 19% (95% CI, -3% to 36%; P=0.08) with transfer for PCI. CONCLUSIONS: Even when transfer to an angioplasty center is necessary, primary PCI remains superior to immediate thrombolysis. Organization of ambulance systems, prehospital management, and adequate PCI capacity appear now to be the key issues in providing reperfusion therapy for AMI. PMID- 14530208 TI - D-cycloserine facilitates synaptic plasticity but impairs glutamatergic neurotransmission in rat hippocampal slices. AB - 1. The glycine-binding site of the glutamatergic N-methyl-d-aspartate receptor subtype (NMDAr) has been proposed as a putative target for treating cognitive impairments in neurodegenerative disorders and schizophrenia. Although behavioural evidence has been accumulated showing that the partial agonist d cycloserine (DCS) facilitated learning and memory, physiological mechanisms of the drug still remained to be characterized. In the present study, we have investigated the effects of DCS on glutamatergic neurotransmission and synaptic plasticity in CA1 region of rat hippocampal slices, using extracellular field excitatory postsynaptic potentials. 2. We showed that DCS facilitated NMDAr mediated synaptic potentials. In addition, we found that the magnitude of NMDAr dependent long-term depression was significantly enhanced by the agonist, while the threshold for the induction of lasting potentiations was lowered. 3. We found that DCS decreased neurotransmission mediated by alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA)/kainate subtypes of glutamate receptors. This inhibition was not prevented by the gamma-aminobutyric acid GABAA antagonist bicuculline, but was antagonized by the glycine antagonist strychnine. 4. These results, therefore, show opposite effects of DCS on NMDA and non-NMDA synaptic responses within the hippocampus. They also demonstrate that DCS facilitates long term synaptic plasticity that may support the DCS-induced enhanced cognitive performances. PMID- 14530209 TI - Activity of chlormethiazole at human recombinant GABA(A) and NMDA receptors. AB - 1. Investigation into the modulatory effects of chlormethiazole at human recombinant gamma-aminobutyric acid A receptor (GABAA) and N-methyl-d-aspartate (NMDA) receptors was undertaken to gain insight into its mechanism of action and determine if the drug exhibited any subtype-selective activity. 2. Despite a structural similarity to the beta-subunit-selective compound loreclezole, chlormethiazole did not show any difference in maximum efficacy and only a slight difference in EC50 in its potentiating action at alpha1beta1gamma2 and alpha1beta2gamma2 GABAA receptor subtypes with preference for alpha1beta1gamma2. 3. Similar to the previously reported subtype-dependent activity of pentobarbital, chlormethiazole elicited a significantly greater degree of maximum potentiation on receptors lacking a gamma2 subunit, and also those receptors containing an alpha4 or alpha6 subunit. This also demonstrates that chlormethiazole does not act via the benzodiazepine binding site. 4. Unlike pentobarbital and propofol, chlormethiazole elicited only a slight direct GABAA receptor activation at concentrations up to 1 mm. In addition, the drug did not potentiate anaesthetic-mediated currents elicited by pentobarbital or propofol, suggesting that chlormethiazole may be acting via an anaesthetic binding site. 5. Chlormethiazole produced weak nonselective inhibition of human NMDA NR1a+NR2A and NR1a+NR2B receptors. IC50's were approximately 500 microm that likely exceed the therapeutic dose range for chlormethiazole, indicating that the primary mechanism of the compounds in vivo activity is via GABAA receptors. PMID- 14530210 TI - Effects of acute treatment with paroxetine, citalopram and venlafaxine in vivo on noradrenaline and serotonin outflow: a microdialysis study in Swiss mice. AB - 1. This study investigated whether a single administration of a range of doses (1, 4 and 8 mg kg-1, i.p.) of paroxetine, citalopram or venlafaxine may simultaneously increase extracellular levels of 5-HT ([5-HT]ext) and noradrenaline ([NA]ext) by using in vivo microdialysis in the frontal cortex (FCx) of awake, freely moving Swiss mice. 2. In vivo, paroxetine induced similar increases in cortical [5-HT]ext at the three doses tested, and induced a statistically significant increase in cortical [NA]ext at 4 and 8 mg x kg-1. Citalopram increased neither [5-HT]ext nor [NA]ext at the lowest dose, but increased both neurotransmitter levels at 4 and 8 mg x kg-1. At these doses, citalopram induced greater increases in cortical [5-HT]ext than in [NA]ext. Venlafaxine increased [5-HT]ext and [NA]ext to about 400 and 140% of the respective basal values at 8 mg kg-1. 3. Citalopram and paroxetine have the highest potency to increase cortical [5-HT]ext and [NA]ext, respectively. In addition, the rank of order of efficacy of these antidepressant drugs to increase [5-HT]ext in vivo in the FCx of mice was as follows: venlafaxine>citalopram>paroxetine, while the efficacy to increase cortical [NA]ext in mice of paroxetine and citalopram is similar, and greater than that of venlafaxine. 4. In conclusion, extracellular levels of cortical [NA]ext increase with the highest doses of the very selective SSRI citalopram, as well as with the very potent SSRI paroxetine. Surprisingly, the SNRI venlafaxine increased cortical [5-HT]ext to a greater extent rather than [NA]ext in the range of doses studied in mice. PMID- 14530211 TI - Role of sarcoplasmic reticulum in control of membrane potential and nitrergic response in opossum lower esophageal sphincter. AB - 1. We previously demonstrated that a balance of Ca2+-activated Cl- current (ICl(Ca)) and K+ current activity sets the resting membrane potential of opossum lower esophageal sphincter (LES) circular smooth muscle at approximately -41 mV, which leads to continuous spike-like action potentials and the generation of basal tone. Ionic mechanisms underlying this basal ICl(Ca) activity and its nitrergic regulation remain unclear. Recent studies suggest that spontaneous Ca2+ release from sarcoplasmic reticulum (SR) and myosin light chain kinase (MLCK) play important roles. The current study investigated this possibility. Conventional intracellular recordings were performed on circular smooth muscle of opossum LES. Nerve responses were evoked by electrical square wave pulses of 0.5 ms duration at 20 Hz. 2. In the presence of nifedipine (1 microm), substance P (1 microm), atropine (3 microm) and guanethidine (3 microm), intracellular recordings demonstrated a resting membrane potential (MP) of -38.1+/-0.7 mV (n=25) with spontaneous membrane potential fluctuations (MPfs) of 1-3 mV. Four pulses of nerve stimulation induced slow inhibitory junction potentials (sIJPs) with an amplitude of 6.1+/-0.3 mV and a half-amplitude duration of 1926+/-147 ms (n=25). 3. 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), a specific guanylyl cyclase inhibitor, abolished sIJPs, but had no effects on MPfs. Caffeine, a ryanodine receptor agonist, hyperpolarized MP and abolished sIJPs and MPfs. Ryanodine (20 microm) inhibited the sIJP and induced biphasic effects on MP, an initial small hyperpolarization followed by a large depolarization. sIJPs and MPfs were also inhibited by cyclopiazonic acid, an SR Ca2+ ATPase inhibitor. Specific ICl(Ca) and MLCK inhibitors hyperpolarized the MP and inhibited MPfs and sIJPs. 4. These data suggest that (1). spontaneous release of Ca2+ from the SR activates ICl(Ca), which in turn contributes to resting membrane potential; (2). MLCK is involved in activation of ICl(Ca); (3). inhibition of ICl(Ca) is likely to underlie sIJPs induced by nitrergic innervation. PMID- 14530212 TI - Functional evidence that ATP or a related purine is an inhibitory NANC neurotransmitter in the mouse jejunum: study on the identity of P2X and P2Y purinoceptors involved. AB - 1. Conflicting views exist on whether ATP is a neurotransmitter in the enteric nervous system. We investigated the role of ATP in enteric transmission in circular muscle strips of the mouse jejunum. 2. On PGF2alpha-precontracted muscle strips and in the presence of atropine and guanethidine, electrical field stimulation (EFS, 1-8 Hz) of nonadrenergic noncholinergic (NANC) nerves induced transient relaxations that were abolished by the nerve-conductance blocker tetrodotoxin. The NO synthase blocker l-nitroarginine (l-NOARG) partially inhibited the NANC relaxations to EFS, but fast-twitch relaxations to EFS were still observed in the presence of l-NOARG. 3. In the presence of l-NOARG, ATP, the P2X receptor agonist alphabetaMeATP and the P2Y receptor agonist ADPbetaS relaxed jejunal muscle strips. Tetrodotoxin did not affect the relaxation to ATP and ADPbetaS, but inhibited that to alphabetaMeATP. 4. The l-NOARG-resistant NANC relaxations to EFS were almost abolished by apamin, a blocker of small conductance Ca2+ activated K+ channels, and by suramin and PPADS, blockers of P2 purinoceptors. Relaxations to ATP were almost abolished by apamin and suramin but not affected by PPADS. 5. Desensitisation of alphabetaMeATP-sensitive P2X receptors, the P2X receptor blocker Evans blue and the P2X1,2,3 receptor blocker NF 279 inhibited the l-NOARG-resistant NANC relaxations to EFS and that to alphabetaMeATP without affecting the relaxation to ADPbetaS. Brilliant blue G, a P2X2,5,7 receptor blocker, did not affect the relaxations to EFS. 6. Desensitisation of P2Y receptors and MRS 2179, a P2Y1 receptor blocker, virtually abolished the l-NOARG-resistant NANC relaxations to EFS and the relaxation to ADPbetaS without affecting the relaxation to alphabetaMeATP. 7. Dipyridamole, an adenosine uptake inhibitor, or theophylline and 8-phenyltheophylline, blockers of P1 and A1 purinoceptors, respectively, did not affect the purinergic NANC relaxations to EFS. 8. Our results suggest that ATP or a related purine acts as an inhibitory NANC neurotransmitter in the mouse jejunum, activating P2 but not P1 purinoceptors. Relaxations to the purinergic NANC neurotransmitter mainly involve P2Y receptors of the P2Y1 subtype that are located postjunctionally. Purinergic NANC neurotransmission also involves P2X receptors, most likely of the P2X1 and P2X3 subtype, located pre- and/or postjunctionally. PMID- 14530213 TI - Effects of endomorphin on substantia gelatinosa neurons in rat spinal cord slices. AB - 1. Whole-cell patch recordings were made from substantia gelatinosa (SG) neurons in transverse lumbar spinal cord slices of 15- to 30-day-old rats. 2. Endomorphin 1 (EM-1) or EM-2 (or =+30 mV than at +10 mV.5 We estimate that nisoldipine is about 30 times less selective for delayed-rectifier K+ channels than for L-type Ca2+ channels in fully polarised guinea-pig ventricular myocytes, and several orders less selective in partially depolarised myocytes. PMID- 14530220 TI - Riluzole inhibits spontaneous Ca2+ signaling in neuroendocrine cells by activation of K+ channels and inhibition of Na+ channels. AB - The neuroprotective drug riluzole has multiple effects on cellular signaling. We found that riluzole rapidly and reversibly inhibited spontaneous Ca2+ oscillations in both immortalized GnRH-secreting hypothalamic neurons (GT1 cells) and in the prolactin and growth-hormone-secreting GH3 cell line. At lower concentrations (100 nm-5 microM), riluzole reduced the amplitude and frequency of spontaneous Ca2+ oscillations, whereas at higher concentrations it abolished spontaneous Ca2+ signaling. Whole-cell current clamp recordings in GH3 cells revealed that riluzole decreased the action potential frequency, amplitude, and duration. Riluzole inhibited voltage-gated Na+ currents, increased iberiotoxin sensitive voltage-gated K+ currents, and had no effect on voltage-gated Ca2+ currents in GH3 cells. Riluzole also inhibited voltage-gated Na+ currents and increased voltage-gated K+ channels in GT1 cells. The inhibitory effects of riluzole on Ca2+ signaling were blocked by pretreatment with iberiotoxin in GH3 cells, but only partially reduced by iberiotoxin in GT1 cells. These results indicate that riluzole inhibits Ca2+ signaling primarily by activation of K+ channels in GH3 cells, and also by inhibition of Na+ channels in GT1 cells. Riluzole's inhibition of spontaneous excitability and Ca2+ signaling may be involved in its multiple effects on cellular function in the nervous system. PMID- 14530221 TI - Rho-kinase inhibitor inhibits both myosin phosphorylation-dependent and independent enhancement of myofilament Ca2+ sensitivity in the bovine middle cerebral artery. AB - The role of Rho kinase in Ca2+ sensitization of the contractile apparatus in smooth muscle was investigated in the bovine middle cerebral artery. U46619, a thromboxane A2 analog, induced a greater sustained contraction with a smaller [Ca2+]i elevation than that seen with 118 mm K+. The level of myosin light chain (MLC) phosphorylation obtained in the initial phase of the contraction was higher than that seen with 118 mm K+; thereafter, it gradually declined to a comparable level in the late phase. During the steady state of the U46619-induced contraction, Y27632 (10 microM), a Rho-kinase inhibitor, partially inhibited [Ca2+]i, although it substantially inhibited tension and MLC phosphorylation. Wortmannin (10 microM), an MLC kinase inhibitor, had no significant effect on [Ca2+]i, but it completely inhibited MLC phosphorylation and partially inhibited tension. The wortmannin-resistant tension development was thus not associated with MLC phosphorylation, and this component was completely inhibited by Y27632. In conclusion, U46619 enhanced Ca2+ sensitivity in a manner both dependent and independent of MLC phosphorylation in the bovine middle cerebral artery. Both mechanisms of Ca2+ sensitization can be inhibited by the Rho-kinase inhibitor. PMID- 14530222 TI - Functional role of angiotensin II AT2 receptor in modulation of AT1 receptor mediated contraction in rat uterine artery: involvement of bradykinin and nitric oxide. AB - The aim of the present study was to explore the mechanisms underlying angiotensin II AT2 receptor modulation of AT1 receptor-mediated vasoconstriction in the rat isolated uterine artery, since previous studies have suggested that AT2 receptors may oppose AT1 receptor-mediated effects. Segments of uterine artery were obtained from Sprague-Dawley rats and mounted in small vessel myographs. Concentration-response (CR) curves to angiotensin II (0.1 nm-0.1 microM) were constructed in the absence and presence of PD 123319 (AT2 antagonist; 1 microM), HOE 140 (bradykinin B2 antagonist; 0.1 microM), Nomega-nitro-l-arginine (NOLA) (NOS inhibitor; 30 microM), as well as combinations of these inhibitors. Contractile responses to angiotensin II were expressed as a percent of the response to a K+ depolarizing solution. PD 123319 (1 microM) potentiated angiotensin II-induced contractions; reflected by a significant four-fold leftward shift of the angiotensin II CR curve. HOE 140 (0.1 microM) significantly increased the pEC50 of the angiotensin II CR curve. The combination of HOE 140 plus PD 123319 did not produce additive potentiation. NOLA (30 microM) significantly enhanced sensitivity to angiotensin II, seen as a five-fold leftward shift of the curve, and an augmented maximum contractile response. Combinations of PD 123319 (1 microM) plus NOLA, and of HOE 140 (0.1 microM) plus NOLA, both induced a similar magnitude of potentiation. Cyclic GMP measurements confirmed angiotensin II-induced activation of the nitric oxide (NO) pathway. In conclusion, AT2 receptor-mediated inhibition of angiotensin II-induced contraction of the rat uterine artery involves NO production; a component of which occurs through a bradykinin B2 receptor pathway. PMID- 14530224 TI - Gastrointestinal tolerability and effectiveness of rofecoxib versus naproxen in the treatment of osteoarthritis: a randomized, controlled trial. AB - BACKGROUND: Gastrointestinal (GI) toxicity mediated by dual cyclooxygenase (COX) 1 and COX-2 inhibition of nonsteroidal anti-inflammatory drugs (NSAIDs) can cause serious alterations of mucosal integrity or, more commonly, intolerable GI symptoms that may necessitate discontinuation of therapy. Unlike NSAIDs, rofecoxib targets only the COX-2 isoform. OBJECTIVE: To assess the tolerability of rofecoxib compared with naproxen for treatment of osteoarthritis. DESIGN: Randomized, controlled trial. SETTING: 600 office and clinical research sites. PATIENTS: 5557 patients (mean age, 63 years) with a baseline diagnosis of osteoarthritis of the knee, hip, hand, or spine. INTERVENTION: Rofecoxib, 25 mg/d, or naproxen, 500 mg twice daily. Use of routine medications, including aspirin, was permitted. MEASUREMENTS: Discontinuation due to GI adverse events (primary end point) and use of concomitant medication to treat GI symptoms (secondary end point). Efficacy was determined by patient-reported global assessment of disease status and the Australian/Canadian Osteoarthritis Hand Index, as well as discontinuations due to lack of efficacy. Patients were evaluated at baseline and at weeks 6 and 12. RESULTS: Rates of cumulative discontinuation due to GI adverse events were statistically significantly lower in the rofecoxib group than in the naproxen group (5.9% vs. 8.1%; relative risk, 0.74 [95% CI, 0.60 to 0.92]; P = 0.005), as were rates of cumulative use of medication to treat GI symptoms (9.1% vs. 11.2%; relative risk, 0.79 [CI, 0.66 to 0.96]; P = 0.014]). Subgroup analysis of patients who used low-dose aspirin (13%) and those who previously discontinued using arthritis medication because of GI symptoms (15%) demonstrated a relative risk similar to the overall sample for discontinuation due to GI adverse events (relative risk, 0.56 [CI, 0.31 to 1.01] and 0.53 [CI, 0.34 to 0.84], respectively). No statistically significant difference was observed between treatments for efficacy in treating osteoarthritis or for occurrence of other adverse events. CONCLUSIONS: In patients with osteoarthritis treated for 12 weeks, rofecoxib, 25 mg/d, was as effective as naproxen, 500 mg twice daily, but had statistically significantly superior GI tolerability and led to less use of concomitant GI medications. Benefits of rofecoxib in subgroup analyses were consistent with findings in the overall sample. PMID- 14530223 TI - Angiotensin AT2 receptors: cardiovascular hope or hype? PMID- 14530225 TI - Magnetic resonance cholangiopancreatography: a meta-analysis of test performance in suspected biliary disease. AB - BACKGROUND: Magnetic resonance cholangiopancreatography (MRCP) is one of many newer noninvasive tests that can image the biliary tree. PURPOSE: To precisely estimate the overall sensitivity and specificity of MRCP in suspected biliary obstruction and to evaluate clinically important subgroups. DATA SOURCES: MEDLINE search (January 1987 to March 2003) for studies in English or French, bibliographies, and subject matter experts. STUDY SELECTION: Studies were included if they allowed construction of 2x2 contingency tables of MRCP compared with a reasonable gold standard for at least 1 of the following: the presence, level, or cause of biliary obstruction. DATA EXTRACTION: Two independent observers graded study quality, which included consecutive enrollment, blinding, use of a single (versus composite) gold standard, and nonselective use of the gold standard. Logistic regression was used to examine the influence of publication year, quality score, proportion of patients having a "direct" gold standard, and clinical context on diagnostic performance. DATA SYNTHESIS: Of 498 studies identified, 67 were included (4711 patients). Mixed-effect models were used to estimate the sensitivity and specificity, and quantitative receiver operating characteristic analysis was performed. Magnetic resonance cholangiopancreatography had a high overall pooled sensitivity (95% [+/-1.96 SD: spread of SD, 75% to 99%]) and specificity (97% [spread of SD, 86% to 99%]). The procedure was less sensitive for stones (92%; odds ratio, 0.51 [CI, 0.35 to 0.75]) and malignant conditions (88%; odds ratio, 0.28 [CI, 0.18 to 0.44]) than for the presence of obstruction. In addition, diagnostic performance was higher in studies that were larger, did not use consecutive enrollment, and did not use gold standard assessment for some patients. CONCLUSIONS: Magentic resonance cholangiopancreatography is a noninvasive imaging test with excellent overall sensitivity and specificity for demonstrating the level and presence of biliary obstruction; however, it seems less sensitive for detecting stones or differentiating malignant from benign obstruction. PMID- 14530226 TI - Racial differences pertaining to a belief about lung cancer surgery: results of a multicenter survey. AB - BACKGROUND: Patients at the Philadelphia Veterans Affairs Medical Center frequently voice concern that air exposure during lung cancer surgery might cause tumor spread. Several African-American patients asserted that this belief was common in the African-American community. OBJECTIVE: To assess the prevalence of the belief that air exposure during lung cancer surgery might cause tumor spread and gauge the influence of this belief on the willingness of African-American and white patients to have lung cancer surgery. DESIGN: Prospective questionnaire survey. SETTING: Philadelphia Veterans Affairs Medical Center and University of Pennsylvania, Philadelphia, Pennsylvania; Los Angeles Veterans Affairs Medical Center, Los Angeles, California; and Medical University of South Carolina, Charleston, South Carolina. PATIENTS: 626 consecutive patients in pulmonary and lung cancer clinics. MEASUREMENTS: None. RESULTS: 38% of patients (61% of whom were African American and 29% of whom were white) stated that they believe air exposure at surgery causes tumor spread. The most significant predictor of belief was African-American race (odds ratio, 3.5 [95% CI, 1.9 to 6.5]), even after controlling for other relevant variables in a multivariable analysis. Nineteen percent of African Americans stated that this belief was a reason for opposing surgery, and 14% would not accept their physicians' assertion that the belief is false. These rates were also statistically significantly higher among African American than white patients. CONCLUSIONS: Belief in accelerated tumor spread at surgery is prevalent among general pulmonary outpatients and lung cancer clinic patients facing lung surgery, particularly among African-American patients. Our findings may pertain to key racial disparities in lung cancer surgery and survival rates and suggest that culturally sensitive physician training or outreach programs directed at disparate beliefs and attitudes may help to address racial discrepancies in health care outcomes. PMID- 14530227 TI - An outbreak of severe acute respiratory syndrome among hospital workers in a community hospital in Hong Kong. AB - BACKGROUND: During outbreaks, hospital workers are at high risk for nosocomial infection with severe acute respiratory syndrome (SARS)-associated coronavirus. OBJECTIVE: To examine how hospital workers became infected and whether they transmit the virus to their families. DESIGN: Retrospective descriptive study. SETTING: 529-bed community hospital in Hong Kong. PATIENTS: 40 hospital workers infected with SARS-associated coronavirus over a 6-week period (25 March through 5 May 2003). MEASUREMENTS: Percentage of infected hospital workers according to job category. RESULTS: The cumulative incidence was highest among health care assistants, followed by physicians and nurses (8%, 5%, and 4%, respectively). Most hospital workers were infected from direct contact with patients with SARS, who primarily were in general wards and had unsuspected infection. At the time of contact, all hospital workers had used masks but not necessarily other protective devices. Affected hospital workers did not infect their families. CONCLUSION: Before isolation of all patients with clinically confirmed or suspected SARS, routine use of several protective devices, and training of staff in infection control, many health care workers were infected with SARS from patients with unsuspected cases. PMID- 14530228 TI - Innovations in access to care: a patient-centered approach. AB - To receive health care, patients with nonemergent problems must gain access to a complex, interdependent ambulatory care system currently structured around the conventional office appointment model. The system does not effectively accommodate diverse patient needs and preferences, contributing to both overuse and underuse of health care resources. A patient-centered access model would help patients secure appropriate and preferred medical assistance when and where it is needed. Characteristics of patient-centered access include availability, appropriateness, preference, and timeliness. One or more of these characteristics often is missing in patients' health care experiences. The goal of this paper is to present patient-centered access as an integrated concept and philosophy to provide context for evaluating specific access initiatives. On the basis of an assessment of existing literature, 3 organizing principles of patient-centered access are proposed and discussed: work at the high end of expertise; align care with need and preference; and serve when service is needed. Patient-centered access warrants serious consideration, given the stakes involved for patients, providers, and payers. Few concepts support all 6 of the Institute of Medicine's aims for the 21st century: safety, effectiveness, patient-centeredness, timeliness, efficiency, and equitability. Patient-centered access is such a concept. PMID- 14530229 TI - Evaluation of acute knee pain in primary care. AB - BACKGROUND: The evaluation of acute knee pain often includes radiography of the knee. OBJECTIVE: To synthesize the literature to determine the role of radiologic procedures in evaluating common causes of acute knee pain: fractures, meniscal or ligamentous injuries, osteoarthritis, and pseudogout. DATA SOURCES: MEDLINE search from 1966 to October 2002. STUDY SELECTION: We included all published, peer-reviewed studies of decision rules for fractures. We included studies that used arthroscopy as the gold standard for measuring the accuracy of the physical examination and magnetic resonance imaging (MRI) for meniscal and ligamentous knee damage. We included all studies on the use of radiographs in pseudogout. DATA EXTRACTION: We extracted all data in duplicate and abstracted physical examination and MRI results into 2 x 2 tables. DATA SYNTHESIS: Among the 5 decision rules for deciding when to use plain films in knee fractures, the Ottawa knee rules (injury due to trauma and age >55 years, tenderness at the head of the fibula or the patella, inability to bear weight for 4 steps, or inability to flex the knee to 90 degrees) have the strongest supporting evidence. When the history suggests a potential meniscal or ligamentous injury, the physical examination is moderately sensitive (meniscus, 87%; anterior cruciate ligament, 74%; and posterior cruciate ligament, 81%) and specific (meniscus, 92%; anterior cruciate ligament, 95%; and posterior cruciate ligament, 95%). The Lachman test is more sensitive and specific for ligamentous tears than is the drawer sign. For meniscal tears, joint line tenderness is sensitive (75%) but not specific (27%), while the McMurray test is specific (97%) but not sensitive (52%). Compared with the physical examination, MRI is more sensitive for ligamentous and meniscal damage but less specific. When the differential diagnosis for acute knee pain includes an exacerbation of osteoarthritis, clinical features (age >50 years, morning stiffness <30 minutes, crepitus, or bony enlargement) are 89% sensitive and 88% specific for underlying chronic arthritis. Adding plain films improves sensitivity slightly but not specificity. Plain films for pseudogout are not sensitive or specific, according to limited-quality studies. CONCLUSIONS: We recommend the Ottawa knee rules to decide when to obtain plain films for suspected knee fracture. A careful physical examination should be sufficient to decide whether to refer patients with potential meniscal and ligament injuries, and we prefer clinical criteria rather than plain films for evaluating osteoarthritis. We do not recommend using plain films to diagnose pseudogout. PMID- 14530230 TI - The lessons of SARS. AB - Given the low mortality and morbidity of the severe acute respiratory syndrome (SARS) compared with other public health scourges, is the attention devoted to it misdirected? The SARS experience has provided at least 4 enduring lessons. First, by providing a test of the capacity of each part of the public health system, from national to local and hospital responses, it has better prepared the world for the anticipated and much-feared next real pandemic. Second, SARS has reemphasized that from housing, sexual practices, and slaughtering techniques to health care capacity, the situation in other, especially developing, countries affects us. Global cooperation is necessary not only for justice but to ensure our own health. Third, despite trends toward commercialization, easier lives, and self-centered individualism, the response of health care professionals to SARS reaffirmed dedication to caring for the sick even at great personal risks as the core ethical principle of medicine. Finally, SARS also emphasized the importance of the duty of health care administrators and senior physicians to rapidly institute procedures to maximize the safety of frontline physicians and nurses. These lessons will be valuable far beyond the SARS episode. PMID- 14530231 TI - Listening to SARS: lessons for infection control. PMID- 14530232 TI - General internists and family physicians: partners in geriatric medicine? PMID- 14530233 TI - General internal medicine and geriatrics: collaboration to address the aging imperative can't wait. PMID- 14530234 TI - Echinacea for the common cold. PMID- 14530235 TI - Echinacea for the common cold. PMID- 14530236 TI - Echinacea for the common cold. PMID- 14530237 TI - Echinacea for the common cold. PMID- 14530238 TI - Echinacea for the common cold. PMID- 14530239 TI - Echinacea for the common cold. PMID- 14530240 TI - Alcohol, postmenopausal hormones, and breast cancer. PMID- 14530241 TI - Update in hospital medicine. PMID- 14530242 TI - Acute migraine treatment guideline. PMID- 14530243 TI - An unusual case of ischemic bowel. PMID- 14530244 TI - General internal medicine and geriatrics: building a foundation to improve the training of general internists in the care of older adults. AB - Internists--"doctors for adults"--provide most of the medical care given to older Americans, especially those with serious chronic disease. Nonetheless, the United States lacks an adequate physician workforce with mastery in caring for older persons and with expertise in building knowledge about how best to provide this care. This supplement aims to strengthen the physician workforce by fostering incremental and sustained improvements in the training of internal medicine residents in the care of older adults and in the development of geriatrics oriented general internal medicine faculty. It identifies 3 major barriers to these improvements: lack of adequately trained teachers and mentors, the belief that explicit training in geriatrics has little to offer the generalist, and inadequate funding. Three strategies offer particular promise in overcoming these barriers: engaging directors of internal medicine residency programs, funding centers to promote collaboration between teaching and research programs in general internal medicine and geriatrics, and providing substantial incremental funding on the national level to pay for the time required to care for frail older patients and to teach and do research about this care. The barriers and strategies identified in this supplement may also inform efforts to enhance the skills of practicing physicians and improve training and faculty development in family medicine and other disciplines. PMID- 14530245 TI - Development of geriatrics-oriented faculty in general internal medicine. AB - The need for adequate geriatrics training for the physician workforce has been recognized for decades. However, there are not enough academic geriatricians to provide for the educational needs of trainees, and this situation is not expected to change in the future. General internists are often responsible for teaching medical students and internal medicine residents to care for elderly patients in inpatient and ambulatory settings. These academic general internists could play a pivotal role in providing geriatrics instruction. To characterize what is being done to develop geriatrics-oriented general internal medicine faculty, we identified current practices, "best practices," goals and targets, and barriers to achieving those goals and targets. We reviewed the literature on faculty development programs for general internal medicine faculty, and we held focus groups and structured interviews with general internal medicine unit chiefs and directors of Geriatric Centers of Excellence at 46 medical schools throughout the United States. We found a need for programs to develop geriatrics-oriented academic general internists. Although general internal medicine faculties seem receptive to further geriatrics training, important obstacles exist. These include inadequate time and resources as well as motivational and attitudinal challenges. We discuss potential solutions for overcoming these barriers and the implications of these solutions for stakeholders. PMID- 14530246 TI - Geriatrics training in general internal medicine fellowship programs: current practice, barriers, and strategies for improvement. AB - To ensure its growth and prosperity, general internal medicine will need to embrace care of the elderly, research on aging, and geriatrics education as components of its core mission. Experts agree that general internal medicine fellows could benefit from increased opportunities in research on aging and geriatrics education; however, important barriers will hamper efforts to integrate geriatrics training into general internal medicine fellowship programs. This article reviews the barriers to integration and proposes solutions for overcoming those barriers. As a result of interviews and meetings with a broad representation of general internists, geriatricians, funding agencies, and policymakers, we propose 2 interventions: 1) the development of institutional program grants to foster collaboration between general internal medicine and geriatrics faculty in the training of general internal medicine fellows and 2) the creation of a 3-year fellowship program combining general internal medicine and geriatrics. This article discusses the importance of evaluating these and other programs intended to increase the geriatrics experience of general internal medicine fellows, and it describes the potential implications of these changes for a broad array of stakeholder institutions. PMID- 14530247 TI - Improving geriatrics training in internal medicine residency programs: best practices and sustainable solutions. AB - National surveys indicate a need for additional training in geriatrics during internal medicine residencies. This paper describes 1) "best practices" for integrating geriatrics education into internal medicine residency programs, 2) barriers to implementation of these practices, and 3) possible ways to improve geriatrics training for internal medicine residents. These best practices were determined by a systematic review of the literature and through interviews with leaders of 26 residency and geriatrics programs concerned with geriatrics training for residents. The most successful programs have clinical experiences with 3 key elements: model geriatric care in 1 or more settings (for example, in the hospital or in ambulatory practice), patient care across sites or transitions of care, and interdisciplinary teamwork. Barriers include attitudes, few faculty, need for relationships with nontraditional training sites, and lack of funding. Local solutions include engaging the internal medicine program director to accomplish a mutual goal--for example, by creating a model geriatrics training experience in which residents demonstrate their skill in a new Accreditation Council of Graduate Medical Education competency (such as systems-based practice). National solutions include reaching consensus on the competencies in geriatrics that should be achieved by board-eligible internists. This may mean increasing the number of questions that test geriatrics competency in the certifying and in-training examinations, increasing numbers of faculty members able to teach and model geriatric care, developing "effective medical resident teaching" courses for nonphysician faculty, and lobbying for improved systems of care. PMID- 14530249 TI - Summaries for patients. Noninvasive diagnosis of biliary disease. PMID- 14530248 TI - Summaries for patients. Gastrointestinal side effects of rofecoxib and naproxen. PMID- 14530250 TI - Summaries for patients. Patient beliefs about lung cancer surgery. PMID- 14530251 TI - Summaries for patients. Severe acute respiratory syndrome (SARS) in health care workers. PMID- 14530252 TI - Finding Ping. PMID- 14530253 TI - Priapism induced by proton-pump inhibitors and misoprostol. PMID- 14530254 TI - A single protease, Apg4B, is specific for the autophagy-related ubiquitin-like proteins GATE-16, MAP1-LC3, GABARAP, and Apg8L. AB - Apg8 is a ubiquitin-like protein involved in autophagy in yeast. Apg8 is covalently but transiently attached to membrane lipids through the actions of activating, conjugating, and processing/deconjugating enzymes. The mammalian Apg8 homologues GATE-16, GARARAP, and MAP1-LC3 have been implicated in intra-Golgi transport, receptor sorting, and autophagy, respectively. All are served by a single set of activating and conjugating enzymes. Here we identify a novel mammalian Apg8 homologue, which we name Apg8L, and describe the synthesis of electrophilic probes based on the GATE-16, GARARAP, MAP1-LC3, and Apg8L proteins. These probes not only form specific adducts in crude cell lysates, but also allow identification of the cellular proteases specific for the C termini of these Apg8 homologues. We find a single protease, Apg4B/autophagin-1, capable of acting on GATE-16, GABARAP, MAP1-LC3, and Apg8L. The Apg4B/autophagin-1 protease thus serves as a processing/deconjugating enzyme for these four highly divergent mammalian Apg8 homologues. PMID- 14530255 TI - Mapping the arrestin-receptor interface. Structural elements responsible for receptor specificity of arrestin proteins. AB - Arrestins selectively bind to phosphorylated activated forms of their cognate G protein-coupled receptors. Arrestin binding prevents further G protein activation and often redirects signaling to other pathways. The comparison of the high resolution crystal structures of arrestin2, visual arrestin, and rhodopsin as well as earlier mutagenesis and peptide inhibition data collectively suggest that the elements on the concave sides of both arrestin domains most likely participate in receptor binding directly, thereby dictating its receptor preference. Using comparative binding of visual arrestin/arrestin2 chimeras to the preferred target of visual arrestin, light-activated phosphorylated rhodopsin (PRh*), and to the arrestin2 target, phosphorylated activated m2 muscarinic receptor (P-m2 mAChR*), we identified the elements that determine the receptor specificity of arrestins. We found that residues 49-90 (beta-strands V and VI and adjacent loops in the N-domain) and 237-268 (beta-strands XV and XVI in the C domain) in visual arrestin and homologous regions in arrestin2 are largely responsible for their receptor preference. Only 35 amino acids (22 of which are nonconservative substitutions) in the two elements are different. Simultaneous exchange of both elements between visual arrestin and arrestin2 fully reverses their receptor specificity, demonstrating that these two elements in the two domains of arrestin are necessary and sufficient to determine their preferred receptor targets. PMID- 14530256 TI - Influence of polymerase II processivity on alternative splicing depends on splice site strength. AB - Transcription and pre-mRNA splicing are coordinated temporally and spatially, and both processes can influence each other. In particular, control of transcriptional elongation by RNA polymerase II has proved to be important for alternative splicing regulation. In this report we demonstrate that the efficiency of exon recognition by the splicing machinery is crucial for the elongation control. Alternative splicing of the fibronectin extra domain I (EDI) is because the polypyrimidine tract of its 3'-splice site occurs suboptimal. By mutating the polypyrimidine tract of EDI in two different positions, individually or in combination, and by disrupting its exonic splicing silencer, we managed to generate minigenes with increasing degrees of exon recognition. Improvement of exon recognition is evidenced by independence from the splicing regulator SF2/ASF for inclusion. The mutated minigenes were used to transfect human cells in culture and study the responsiveness of EDI alternative splicing to activation or inhibition of pol II elongation. Our results revealed that responsiveness of exon skipping to elongation is inversely proportional to 3'-splice site strength, which means that the better the alternative exon is recognized by the splicing machinery, the less its degree of inclusion is affected by transcriptional elongation. PMID- 14530257 TI - Antisense depletion of death-associated protein kinase promotes apoptosis. AB - Death-associated protein kinases (DAPK) are serine/threonine protein kinases that have an important role in regulating cell death. In this study two antisense approaches were employed to down-regulate expression of the endogenous DAPK-alpha and DAPK-beta proteins. Transient expression of an antisense DAPK cDNA or antisense morpholino oligonucleotides in HeLa, 3T3, or primary human vascular smooth muscle cells demonstrate that decreased DAPK expression promotes a spontaneous, caspase-mediated apoptosis as evidenced by increased activities of caspases-3 and -9. Clonal HeLa cell lines with attenuated levels of DAPK expression, obtained following selection in the presence of antisense DAPK cDNA, are more sensitive to tumor necrosis factor-induced caspase-mediated apoptosis, and their sensitivity is inversely related to DAPK expression. In contrast, HeLa cells with reduced DAPK expression are moderately resistant to cell death induced by interferon-gamma. This finding is consistent with previous studies showing that DAPK has a role in promoting caspase-independent cell death. Together, these studies demonstrate that the cellular activities of DAPK are critical for antagonizing caspase-dependent apoptosis to promote cell survival under normal cell growth conditions. PMID- 14530258 TI - Membrane localization, oligomerization, and phosphorylation are required for optimal raf activation. AB - Activation of the serine/threonine kinase c-Raf-1 requires membrane localization, phosphorylation, and oligomerization. To study these mechanisms of Raf activation more precisely, we have used a membrane-localized fusion protein, myr-Raf-GyrB, which can be activated by coumermycin-induced oligomerization in NIH3T3 transfectants. By introducing a series of point mutations into the myr-Raf-GyrB kinase domain (S338A, S338A/Y341F, Y340F/Y341F, and T491A/S494A) we can separately study the role that membrane localization, phosphorylation, and oligomerization play in the process of Raf activation. We find that phosphorylation of Ser-338 plays a critical role in Raf activation and that this requires membrane localization but not oligomerization of Raf. Mutation of Tyr 341 had a limited effect, whereas mutation of both Ser-338 and Tyr-341 resulted in a synergistic loss of Raf activation following coumermycin-induced dimerization. Importantly, we found that membrane localization and phosphorylation of Ser-338 were not sufficient to activate Raf in the absence of oligomerization. Thus, our studies suggest that three key steps are required for optimal Raf activation: recruitment to the plasma membrane by GTP-bound Ras, phosphorylation via membrane-resident kinases, and oligomerization. PMID- 14530259 TI - Mi-2 beta associates with BRG1 and RET finger protein at the distinct regions with transcriptional activating and repressing abilities. AB - Mi-2 beta is the main component of the nucleosome remodeling and deacetylase complex and plays an important role in epigenetic transcriptional repression. Here we show that the amino-terminal and carboxyl-terminal regions of Mi-2 beta have distinct transcriptional activities and bind to BRG1, a component of the SWI/SNF complex, and the RET finger protein (RFP), respectively. Analysis by luciferase reporter assay revealed that the amino-terminal region of Mi-2 beta has a strong transactivating ability, whereas its carboxyl-terminal region has transcriptional repressive activity. Co-localization and association of Mi-2, RFP, and histone deacetylase 1 suggested that these proteins cooperate in transcriptional repression. Furthermore, the functional importance of the association of Mi-2 beta and RFP was confirmed by using Rfp-/- fibroblasts. On the other hand, we demonstrated that Mi-2 and BRG1 were associated with each other and that the bromodomain region of BRG1 strongly suppressed transactivation by the amino-terminal region of Mi-2 beta. The findings that Mi-2 beta interacts with both transactivating and repressing proteins and directly associates with another chromatin remodeling protein, BRG1, provide new insight into the formation of multiprotein supercomplex involved in transcriptional regulation. PMID- 14530260 TI - Replication factor C clamp loader subunit arrangement within the circular pentamer and its attachment points to proliferating cell nuclear antigen. AB - Replication factor C (RFC) is a heteropentameric AAA+ protein clamp loader of the proliferating cell nuclear antigen (PCNA) processivity factor. The prokaryotic homologue, gamma complex, is also a heteropentamer, and structural studies show the subunits are arranged in a circle. In this report, Saccharomyces cerevisiae RFC protomers are examined for their interaction with each other and PCNA. The data lead to a model of subunit order around the circle. A characteristic of AAA+ oligomers is the use of bipartite ATP sites in which one subunit supplies a catalytic arginine residue for hydrolysis of ATP bound to the neighboring subunit. We find that the RFC(3/4) complex is a DNA-dependent ATPase, and we use this activity to determine that RFC3 supplies a catalytic arginine to the ATP site of RFC4. This information, combined with the subunit arrangement, defines the composition of the remaining ATP sites. Furthermore, the RFC(2/3) and RFC(3/4) subassemblies bind stably to PCNA, yet neither RFC2 nor RFC4 bind tightly to PCNA, indicating that RFC3 forms a strong contact point to PCNA. The RFC1 subunit also binds PCNA tightly, and we identify two hydrophobic residues in RFC1 that are important for this interaction. Therefore, at least two subunits in RFC make strong contacts with PCNA, unlike the Escherichia coli gamma complex in which only one subunit makes strong contact with the beta clamp. Multiple strong contact points to PCNA may reflect the extra demands of loading the PCNA trimeric ring onto DNA compared with the dimeric beta ring. PMID- 14530261 TI - Alterations in subcellular localization of p38 MAPK potentiates endothelin stimulated COX-2 expression in glomerular mesangial cells. AB - Endothelin-1 (ET-1) is a potent vasoconstrictor peptide with mitogenic actions linked to activation of tyrosine kinase signaling pathways. ET-1 induces cyclooxygenase-2 (COX-2), an enzyme that converts arachidonic acid to pro inflammatory eicosanoids. Activation of each of the three major mitogen-activated protein kinase (MAPK) pathways, ERK1/2, JNK/SAPK, and p38 MAPK (p38), have been shown to enhance the expression of COX-2. Negative regulation of MAPK may occur via a family of dual specificity phosphatases referred to as mitogen-activated protein kinase phosphatases (MKP). The goal of this work was to test the hypothesis that wild type MKP-1 regulates the expression of ET-1-induced COX-2 expression by inhibiting the activation of p38 in cultured glomerular mesangial cells (GMC). An adenovirus expressing both wild type and a catalytically inactive mutant of MKP-1 (MKP-1/CS) were constructed to study ET-1-regulated MAPK signaling and COX-2 expression in cultured GMC. ET-1 stimulated the phosphorylation of ERK and p38 alpha MAPK and induced the expression of COX-2. Expression of COX-2 was partially blocked by U0126, a MEK inhibitor, and SB 203580, a p38 MAPK inhibitor. Adenoviral expression of MKP-1/CS augmented basal and ET-1-induced phosphorylation of p38 alpha MAPK with less pronounced effects on ERK1/2 phosphorylation. Ectopic expression of wild type MKP-1 blocked the phosphorylation of p38 alpha MAPK by ET-1 but increased the phosphorylation of p38 gamma MAPK. Co-precipitation studies demonstrated association of MKP-1 with p38 alpha MAPK and ERK1/2. Immunofluorescent image analysis demonstrated trapping of phospho-p38 MAPK in the cytoplasm by MKP-1/CS/green fluorescent protein. ET-1 stimulated expression of COX-2 was increased in MKP-1/CS versus LacZ or green fluorescent protein-infected control cells. These results indicate that MKP-1 demonstrates a relative selectivity for p38 alpha MAPK versus p38 gamma MAPK in GMC and is likely to indirectly regulate the expression of COX-2. PMID- 14530262 TI - The alpha, but not the beta, isoform of the human thromboxane A2 receptor is a target for nitric oxide-mediated desensitization. Independent modulation of Tp alpha signaling by nitric oxide and prostacyclin. AB - In humans, thromboxane A2 signals through two thromboxane A2 receptor (TP) isoforms termed TP alpha and TP beta. Signaling by TP alpha, but not TP beta, is subject to prostacyclin-induced desensitization mediated by direct protein kinase (PK) A phosphorylation where Ser329 represents the phosphotarget (Walsh, M. T., Foley, J. F., and Kinsella, B. T. (2000) J. Biol. Chem. 275, 20412-20423). In the current study, the effect of the vasodilator nitric oxide (NO) on intracellular signaling by the TP isoforms was investigated. The NO donor 3 morpholinosydnonimine, HCl (SIN-1) and 8-bromo-guanosine 3',5'-cyclic monophosphate (8-Br-cGMP) functionally desensitized U46619-mediated calcium mobilization and inositol 1,4,5-trisphosphate generation by TP alpha whereas signaling by TP beta was unaffected by either agent. NO-mediated desensitization of TP alpha signaling occurred through a PKG-dependent, PKA- and PKC-independent mechanism. TP alpha, but not TP beta, was efficiently phosphorylated by PKG in vitro and underwent NO/PKG-mediated phosphorylation in whole cells. Deletion/site directed mutagenesis and metabolic labeling studies identified Ser331 as the target residue of NO-induced PKG phosphorylation of TP alpha. Although TP alpha S331A was insensitive to NO/PKG-desensitization, similar to wild type TP alpha its signaling was fully desensitized by the prostacyclin receptor agonist cicaprost occurring through a PKA-dependent mechanism. Conversely, signaling by TP alpha S329A was insensitive to cicaprost stimulation whereas it was fully desensitized by NO/PKG signaling. In conclusion, TP alpha undergoes both NO- and prostacyclin-mediated desensitization that occur through entirely independent mechanisms involving direct PKG phosphorylation of Ser331, in response to NO, and PKA phosphorylation of Ser329, in response to prostacyclin, within the unique carboxyl-terminal tail domain of TP alpha. On the other hand, signaling by TP beta is unaffected by either NO or prostacyclin. PMID- 14530263 TI - p57Kip2 regulates actin dynamics by binding and translocating LIM-kinase 1 to the nucleus. AB - p57Kip2 is the only cyclin-dependent kinase (Cdk) inhibitor shown to be essential for mouse embryogenesis. The fact suggests that p57 has a specific role that cannot be compensated by other Cdk inhibitors. LIM-kinase 1 (LIMK-1) is a downstream effector of the Rho family of GTPases that phosphorylates and inactivates an actin depolymerization factor, cofilin, to induce the formation of actin fiber. Here we demonstrate that p57 regulates actin dynamics by binding and translocating LIMK-1 from the cytoplasm into the nucleus, which in turn results in a reorganization of actin fiber. The central region of p57, a unique feature among the Cdk inhibitors, and the N-terminal region of LIMK-1, which contains the LIM domains were essential for the interaction. Expression of p57, but not p27Kip1 or a p57 mutant, with a deletion in the central region was shown to induce marked reorganization of actin filament and a translocation of LIMK-1. Our findings indicate p57 may act as a key regulator in embryogenesis by bearing two distinct functions, the regulation of cell cycle through binding to Cdks and the regulation of actin dynamics through binding to LIMK-1, both of which should be important in developmental procedure. PMID- 14530264 TI - The redox state of the cell regulates the ligand binding affinity of human neuroglobin and cytoglobin. AB - Neuroglobin and cytoglobin reversibly bind oxygen in competition with the distal histidine, and the observed oxygen affinity therefore depends on the properties of both ligands. In the absence of an external ligand, the iron atom of these globins is hexacoordinated. There are three cysteine residues in human neuroglobin; those at positions CD7 and D5 are sufficiently close to form an internal disulfide bond. Both cysteine residues in cytoglobin, although localized in other positions than in human neuroglobin, may form a disulfide bond as well. The existence and position of these disulfide bonds was demonstrated by mass spectrometry and thiol accessibility studies. Mutation of the cysteines involved, or the use of reducing agents to break the S-S bond, led to a decrease in the observed oxygen affinity of human neuroglobin by an order of magnitude. The critical parameter is the histidine dissociation rate, which changes by about a factor of 10. The same effect is observed with human cytoglobin, although to a much lesser extent (less than a factor of 2). These results suggest a novel mechanism for the regulation of oxygen binding; contact with an appropriate electron donor would provoke the release of oxygen. Hence the oxygen affinity would be directly linked to the redox state of the cell. PMID- 14530265 TI - The kinesin family member BimC contains a second microtubule binding region attached to the N terminus of the motor domain. AB - The kinesin family member BimC has a highly positively charged domain of approximately 70 amino acids at the N terminus of the motor domain. Motor domain constructs of BimC were prepared with and without this extra domain to determine its influence. The level of microtubules needed for half saturation of the ATPase of BimC motor domain constructs is reduced by approximately 7000-fold at low ionic strength upon addition of this extra N-terminal extension. Although the change in microtubule affinity is less at higher salt, addition of the N-terminal domain still produces a 20-fold increase in affinity for microtubules in 200 mm potassium acetate. A fusion protein of the N-terminal domain and thioredoxin binds tightly to MTs at low salt, consistent with the increased affinity of motor domain constructs (which contain the N-terminal domain) being due to the additional binding of the N-terminal domain to the microtubule. Hydrodynamic analysis indicates that the N-terminal extension is in a highly extended conformation, suggesting that it may be intrinsically disordered. Fusion of the N terminal extension of BimC onto the motor domain of conventional kinesin produces a similar large increase in microtubule affinity without significant reduction in kcat or velocity in an in vitro motility assay, suggesting that the N-terminal extension can act in a modular manner to increase the microtubule affinity of kinesin motor domains without a decrease in velocity. PMID- 14530266 TI - Enzymatic mechanism of RNA translocation in double-stranded RNA bacteriophages. AB - Many complex viruses acquire their genome by active packaging into a viral precursor particle called a procapsid. Packaging is performed by a viral portal complex, which couples ATP hydrolysis to translocation of nucleic acid into the procapsid. The packaging process has been studied for a variety of viruses, but the mechanism of the associated ATPase remains elusive. In this study, the mechanism of RNA translocation in double-stranded RNA bacteriophages is characterized using rapid kinetic analyses. The portal complex of bacteriophage 8 is a hexamer of protein P4, which exhibits nucleotide triphosphatase activity. The kinetics of ATP binding reveals a two-step process: an initial, fast, second order association, followed by a slower, first-order phase. The slower phase exhibits a high activation energy and has been assigned to a conformational change. ATP binding becomes cooperative in the presence of RNA. Steady-state kinetics of ATP hydrolysis, which proceeds only in the presence of RNA, also exhibits cooperativity. On the other hand, ADP release is fast and RNA independent. The steady-state rate of hydrolysis increases with the length of the RNA substrate indicating processive translocation. Raman spectroscopy reveals that RNA binds to P4 via the phosphate backbone. The ATP-induced conformational change affects the backbone of the bound RNA but leaves the protein secondary structure unchanged. This is consistent with a model in which cooperativity is induced by an RNA link between subunits of the hexamers and translocation is effected by an axial movement of the subunits relative to one another upon ATP binding. PMID- 14530268 TI - An isolated class II aminoacyl-tRNA synthetase insertion domain is functional in amino acid editing. AB - Aminoacyl-tRNA synthetases are responsible for activating specific amino acids and transferring them onto cognate tRNA molecules. Due to the similarity in many amino acid side chains, certain synthetases misactivate non-cognate amino acids to an extent that would be detrimental to protein synthesis if left uncorrected. To ensure accurate translation of the genetic code, some synthetases therefore utilize editing mechanisms to hydrolyze non-cognate products. Previously class II Escherichia coli proline-tRNA synthetase (ProRS) was shown to exhibit pre- and post-transfer editing activity, hydrolyzing a misactivated alanine-adenylate (Ala AMP) and a mischarged Ala-tRNAPro variant, respectively. Residues critical for the editing activity (Asp-350 and Lys-279) are found in a novel insertion domain (INS) positioned between motifs 2 and 3 of the class defining aminoacylation active site. In this work, we present further evidence that INS is responsible for editing in ProRS. We deleted the INS from wild-type E. coli ProRS to yield DeltaINS-ProRS. While DeltaINS-ProRS was still capable of misactivating alanine, the truncated construct was defective in hydrolyzing non-cognate Ala-AMP. When the INS domain was cloned and expressed as an independent protein, it was capable of deacylating a mischarged Ala-microhelixPro variant. Similar to full-length ProRS, post-transfer editing was abolished in a K279A mutant INS. We also show that YbaK, a protein of unknown function from Haemophilus influenzae with high sequence homology to the prokaryotic INS domain, was capable of deacylating Ala tRNAPro and Ala-microhelixPro variants but not cognate Pro-tRNAPro. Thus, we demonstrate for the first time that an independently folded class II synthetase editing domain and a previously identified homolog can catalyze a hydrolytic editing reaction. PMID- 14530267 TI - Inhibitor of myogenic family, a novel suppressor of store-operated currents through an interaction with TRPC1. AB - Depletion of intracellular Ca2+ stores leads to the activation of Ca2+ inflow through store-operated Ca2+ channels. Although the identity of these channels is unknown, there is considerable evidence that the transient receptor potential channel 1 (TRPC1) participates in the formation of these channels. We show that TRPC1 physically interacts with the a-isoform of the inhibitor of the myogenic family (I-mfa), a known inhibitor of basic helix-loop-helix transcription factors, in vitro and in vivo. The interaction is mediated by the C-terminal cytoplasmic tail of TRPC1 and the C-terminal cysteine-rich domain of I-mfa. Using the whole cell configuration of the patch clamp technique, we show that ectopic expression of I-mfa in CHO-K1 cells reduces native store-activated Ca2+ currents, whereas knock-down of endogenous I-mfa in A431 cells by RNA interference enhances these currents. Pipette perfusion of purified recombinant I-mfa rescues the effect of I-mfa knock-down on store-operated conductance. Finally, cell dialysis with a monoclonal antibody specific to TRPC1 results in the suppression of store activated conductance in cells lacking I-mfa, but not in I-mfa expressing cells. We propose that I-mfa functions as a molecular switch to suppress the store dependence of TRPC1. PMID- 14530269 TI - A novel effect of bismuth ions: selective inhibition of the biological activity of glycine-extended gastrin. AB - Although bismuth salts have been used for over two centuries for the treatment of various gastrointestinal disorders, the mechanism of their therapeutic action remains controversial. Because gastrins bind two trivalent ferric ions with high affinity, and because ferric ions are essential for the biological activity of glycine-extended gastrin 17, we have investigated the hypothesis that trivalent bismuth ions influence the biological activity of gastrins. Binding of bismuth ions to gastrins was measured by fluorescence quenching and NMR spectroscopy. The effects of bismuth ions on gastrin-stimulated biological activities were measured in inositol phosphate, cell proliferation, and cell migration assays. Fluorescence quenching experiments indicated that both glycine-extended and amidated gastrin 17 bound two bismuth ions. The NMR spectral changes observed on addition of bismuth ions revealed that Glu-7 acted as a ligand at the first bismuth ion binding site. In the presence of bismuth ions the ability of glycine extended gastrin 17 to stimulate inositol phosphate production, cell proliferation, and cell migration was markedly reduced. In contrast, bismuth ions had little effect on the affinity of the CCK-2 receptor for amidated gastrin 17, or on the stimulation of inositol phosphate production by amidated gastrin 17. We conclude that bismuth ions may act, at least in part, by blocking the effects of glycine-extended gastrin 17 on cell proliferation and cell migration in the gastrointestinal tract. This is the first report of a specific inhibitory effect of bismuth ions on the action of a gastrointestinal hormone. PMID- 14530270 TI - p21-activated kinase-1 signaling mediates cyclin D1 expression in mammary epithelial and cancer cells. AB - p21-activated kinase 1 (Pak1) has been shown recently to induce hyperplasia in the mammary epithelium, a phenotype also manifested by overexpression of cyclin D1, a known indicator of the proliferative stage. Here we investigated the role of the Pak1 pathway in the expression of cyclin D1 using tissue culture models and transgenic mice expressing activated Pak1 in mammary glands. We found that hyperplastic mammary glands from catalytically active Pak1 transgenic mice exhibit a 5- to 7-fold increased expression of cyclin D1 as compared with stage matched wild-type mice. In addition, Pak1 levels were elevated in human breast tumors and also correlated well with increased cyclin D1 expression. Increased expression of Pak1 in breast cancer cells stimulated cyclin D1 promoter activity, elevated levels of cyclin D1 mRNA, protein, and nuclear accumulation of cyclin D1. Conversely, Pak1 inhibition by an auto-inhibitory peptide (amino acids 83 149) or Pak1 knockdown by short interference RNA markedly reduced the expression of cyclin D1, suggesting a requirement of a functional Pak1 pathway for optimal expression of cyclin D1. Results from deletion and mutant analysis indicate that Pak1 regulates cyclin D1 transcription by means of an NF-kappaB-dependent pathway. Together, these findings suggest a model wherein Pak1 regulation of cyclin D1 expression might involve an NF-kappaB-dependent pathway and that hyperplasia in the mammary glands of Pak1-TG mice may be associated, at least in part, with the up-regulation of cyclin D1, and that Pak1 is up-regulated in human breast tumors. PMID- 14530271 TI - Protein profile of tax-associated complexes. AB - Infection with human T-cell leukemia virus type 1 (HTLV-1) results in adult T cell leukemia and HTLV-1-associated myelopathy/tropical spastic paraparesis. Tax, a 40-kDa protein, regulates viral and cellular transcription, host signal transduction, the cell cycle, and apoptosis. Tax has been shown to modulate cellular CREB and NFkappaB pathways; however, to date, its role in binding to various host cellular proteins involved in tumorigenesis has not been fully described. In this study, we describe the Tax-associated proteins and their functions in cells using several approaches. Tax eluted from a sizing column mostly at an apparent molecular mass of 1800 kDa. Following Tax immunoprecipitation, washes with high salt buffer, two-dimensional gel separation, and mass spectrometric analysis, a total of 32 proteins was identified. Many of these proteins belong to the signal transduction and cytoskeleton pathways and transcription/chromatin remodeling. A few of these proteins, including TXBP151, have been shown previously to bind to Tax. The interaction of Tax with small GTPase-cytoskeleton proteins, such as ras GAP1m, Rac1, Cdc42, RhoA, and gelsolin, indicates how Tax may regulate migration, invasion, and adhesion in T-cell cancers. Finally, the physical and functional association of Tax with the chromatin remodeling SWI/SNF complex was assessed using in vitro chromatin remodeling assays, chromatin remodeling factor BRG1 mutant cells, and RNA interference experiments. Collectively, Tax is able to bind and regulate many cellular proteins that regulate transcription and cytoskeletal related pathways, which might explain the pleiotropic effects of Tax leading to T cell transformation and leukemia in HTLV-1-infected patients. PMID- 14530272 TI - Insight into functional aspects of Stt3p, a subunit of the oligosaccharyl transferase. Evidence for interaction of the N-terminal domain of Stt3p with the protein kinase C cascade. AB - Over a decade ago, the gene STT3 was identified in a staurosporine and temperature sensitivity screen of yeast. Subsequently the product of this gene was shown to be a subunit of the endoplasmic reticulum-localized oligosaccharyl transferase (OT) complex. Although stt3 mutants are known to be staurosporine sensitive, we found that mutants of other OT subunits (except ost4 Delta) are staurosporine-resistant, which indicates that this phenotype of stt3 mutants is not simply a consequence of their defect in glycosylation, as previously speculated. Staurosporine sensitivity was found to be an allele-specific phenotype restricted to cells harboring mutations in highly conserved residues in the N-terminal domain of the STT3 protein. Cells bearing mutations in one of the cytosolic-oriented loops (amino acids 158-168) in the N terminus of Stt3p were found to be specifically susceptible to staurosporine. Staurosporine is a specific inhibitor of Pkc1p, and a genetic link had previously been suggested between PKC1 and STT3. It is known that overexpression of PKC1 suppresses the staurosporine sensitivity of the stt3 mutants in an allele-specific manner, which is typical of mutants of Pkc1p cascade. It has been shown that the pkc1 null mutant exhibits lowered OT activity. Our results combined with these previous observations indicate that the N-terminal domain of Stt3p may interact with members of the Pkc1p cascade and consequently mutations in this domain result in staurosporine sensitivity. We further speculate that the Pkc1p regulates OT activity through the N-terminal domain of Stt3p, the C-terminal domain of which possesses the recognition and/or catalytic site of the OT complex. PMID- 14530273 TI - Reproductive fitness and quinone content of Caenorhabditis elegans clk-1 mutants fed coenzyme Q isoforms of varying length. AB - Caenorhabditis elegans clk-1 mutants lack coenzyme Q9 and accumulate the biosynthetic intermediate demethoxy-Q9. A dietary source of ubiquinone (Q) is required for larval growth and development of the gonad and germ cells. We considered that uptake of the shorter Q8 isoform present in the Escherichia coli food may contribute to the Clk phenotypes of slowed development and reduced brood size observed when the animals are fed Q-replete E. coli. To test the effect of isoprene tail length, N2 and clk-1 animals were fed E. coli engineered to produce Q7, Q8, Q9, or Q10. Wild-type nematodes showed no change in reproductive fitness regardless of the Qn isoform fed. clk-1(e2519) fed the Q9 diet showed increased egg production; however, this diet did not improve reproductive fitness of the clk-1(qm30) animals. Furthermore, animals with the more severe clk-1(qm30) allele become sterile and their progeny inviable when fed Q7-containing bacteria. The content of Q7 in the mitochondria of clk-1 animals was decreased relative to Q8, suggesting less effective transport of Q7 to the mitochondria, impaired retention, or decreased stability. Additionally, regardless of E. coli diet, clk 1(qm30) animals contain a dysfunctional dense form of mitochondria. The gonads of clk-1(qm30) worms fed Q7-containing food were severely shrunken and disordered. The differential fertility of clk-1 mutant nematodes fed Q isoforms may result from changes in Q localization, altered recognition by Q-binding proteins, and/or potential defects in mitochondrial function resulting from the mutant CLK-1 polypeptide itself. PMID- 14530274 TI - The AP-2 complex is excluded from the dynamic population of plasma membrane associated clathrin. AB - Numerous biologically relevant substrates are selectively internalized via clathrin-mediated endocytosis. At the plasma membrane the AP-2 complex plays a major role in clathrin coat formation, interacting with both cargo and clathrin. Utilizing simultaneous dual-channel total internal reflection fluorescence microscopy we have analyzed components of the AP-2 complex (alpha- and beta 2 adaptin) during clathrin-mediated endocytosis. Although in static images enhanced green fluorescent protein-tagged AP-2 markers significantly co-localized with clathrin and other components of clathrin-coated pits, AP-2 did not seem to be present in clathrin spots that appeared to undergo internalization or motility parallel to the plane of the plasma membrane. Two populations of clathrin at the plasma membrane seem to exist, the dynamic and the static, and AP-2 appears to be only found within the latter. These results suggest that colocalized clathrin/AP 2 puncta may represent loci for coated pit production and that previous models that assumed AP-2 was retained within clathrin coats during endocytosis may need to be re-evaluated. PMID- 14530275 TI - Apo-calmodulin binds with its C-terminal domain to the N-methyl-D-aspartate receptor NR1 C0 region. AB - Calmodulin (CaM) is the major Ca2+ sensor in eukaryotic cells. It consists of four EF-hand Ca2+ binding motifs, two in its N-terminal domain and two in its C terminal domain. Through a negative feedback loop, CaM inhibits Ca2+ influx through N-methyl-D-aspartate-type glutamate receptors in neurons by binding to the C0 region in the cytosolic tail of the NR1 subunit. Ca2+ -depleted (apo)CaM is pre-associated with a variety of ion channels for fast and effective regulation of channel activities upon Ca2+ influx. Using the NR1 C0 region for fluorescence and circular dichroism spectroscopy studies we found that not only Ca2+ -saturated CaM but also apoCaM bound to NR1 C0. In vitro interaction assays showed that apoCaM also binds specifically to full-length NR1 solubilized from rat brain and to the complete C terminus of the NR1 splice form that contains the C0 plus C2' domain. The Ca2+ -independent interaction of CaM was also observed with the isolated C-but not N-terminal fragment of calmodulin in the independent spectroscopic assays. Fluorescence polarization studies indicated that apoCaM associated via its C-terminal domain with NR1 C0 in an extended conformation and collapsed to adopt a more compact conformation of faster rotational mobility in its complex with NR1 C0 upon addition of Ca2+. Our results indicate that apoCaM is associated with NR1 and that the complex of CaM bound to NR1 C0 undergoes a dramatic conformational change when Ca2+ binds to CaM. PMID- 14530276 TI - Physical and functional interaction of Acyl-CoA-binding protein with hepatocyte nuclear factor-4 alpha. AB - Although acyl-CoA-binding protein (ACBP) has been detected in the nucleus, the physiological significance of this observation is unknown. As shown herein for the first time, ACBP in the nucleus physically and functionally interacted with hepatocyte nuclear factor-4 alpha (HNF-4 alpha), a nuclear binding protein that regulates transcription of genes involved in both lipid and glucose metabolism. Five lines of evidence showed that ACBP bound HNF-4 alpha in vitro and in the nucleus of intact cells. (i) ACBP interaction with HNF-4 alpha elicited significant changes in secondary structure. (ii) ACBP and HNF-4 alpha were coimmunoprecipitated by antibodies to each protein. (iii) Double immunolabeling and laser scanning confocal microscopy (LSCM) of rat hepatoma cells and transfected COS-7 cells significantly colocalized ACBP and HNF-4 alpha within the nucleus and in the perinuclear region close to the nuclear membrane. (iv) LSCM fluorescence resonance energy transfer determined an intermolecular distance of 53 A between ACBP and HNF-4 alpha in rat hepatoma cell nuclei. (v) Immunogold electron microscopy detected ACBP within 43 A of HNF-4 alpha. These interactions were specific since ACBP did not interact with Sp1 or glucocorticoid receptor in these assays. The functional significance of ACBP interaction with HNF-4 alpha was evidenced by mammalian two-hybrid and transactivation assays. ACBP overexpression in COS-7 or rat hepatoma cells enhanced transactivation of an HNF 4 alpha-dependent luciferase reporter plasmid by 3.2- and 1.6-fold, respectively. In contrast, cotransfection with antisense ACBP expression vector inhibited transactivation. LSCM of the individual triple fluorescent-labeled (HNF-4 alpha, ACBP, and luciferase) rat hepatoma cells showed a high correlation (r2, 0.936) between the level of luciferase and the level of ACBP expression. In summary, ACBP physically interacted with HNF-4 alpha in vitro and in intact cells, although ACBP expression level directly correlated with HNF-4 alpha-mediated transactivation in individual cells. PMID- 14530277 TI - The endoplasmic reticulum (ER) translocon can differentiate between hydrophobic sequences allowing signals for glycosylphosphatidylinositol anchor addition to be fully translocated into the ER lumen. AB - The signal sequence within polypeptide chains that designates whether a protein is to be anchored to the membrane by a glycosylphosphatidylinositol (GPI) anchor is characterized by a carboxyl-terminal hydrophobic domain preceded by a short hydrophilic spacer linked to the GPI anchor attachment (omega) site. The hydrophobic domain within the GPI anchor signal sequence is very similar to a transmembrane domain within a stop transfer sequence. To investigate whether the GPI anchor signal sequence is translocated across or integrated into the endoplasmic reticulum membrane we studied the translocation, GPI anchor addition, and glycosylation of different variants of a model GPI-anchored protein. Our results unequivocally demonstrated that the hydrophobic domain within a GPI signal cannot act as a transmembrane domain and is fully translocated even when followed by an authentic charged cytosolic tail sequence. However, a single amino acid change within the hydrophobic domain of the GPI-signal converts it into a transmembrane domain that is fully integrated into the endoplasmic reticulum membrane. These results demonstrated that the translocation machinery can recognize and differentiate subtle changes in hydrophobic sequence allowing either full translocation or membrane integration. PMID- 14530280 TI - Differential activation of a C/EBP beta isoform by a novel redox switch may confer the lipopolysaccharide-inducible expression of interleukin-6 gene. AB - C/EBP beta, a member of the CCAAT/enhancer binding protein (C/EBP) family, is one of the key transcription factors responsible for the induction of a wide array of genes, some of which play important roles in innate immunity, inflammatory response, adipocyte and myeloid cell differentiation, and the acute phase response. Three C/EBP beta isoforms (i.e. LAP*, LAP, and LIP) were known to arise from differential translation initiation and display different functions in gene regulation. C/EBP beta is known to induce interleukin (IL)-6 gene when P388D1 cells are treated with lipopolysaccharide (LPS). Exactly how the transcriptional activities of C/EBP beta isoforms are involved in the regulation of the IL-6 gene remains unclear. Here we report that LPS-induced expression of IL-6 gene in P388D1 cells is mediated by a redox switch-activated LAP*. The intramolecular disulfide bonds of LAP* and LAP have been determined. Among the cysteine residues, amino acid 11 (Cys11) of LAP* plays key roles for determining the overall intramolecular disulfide bonds that form the basis for redox switch regulation. The DNA binding activity and transcriptional activity of LAP* are enhanced under reducing condition. LAP and LIP, lacking 21 and 151 amino acids, respectively, in the N-terminal region, are not regulated in a similar redox responsive manner. Our results indicate that LAP* is the primary isoform of C/EBP beta that regulates, through a redox switch, the LPS-induced expression of the IL 6 gene. PMID- 14530278 TI - Cholesterol depletion results in site-specific increases in epidermal growth factor receptor phosphorylation due to membrane level effects. Studies with cholesterol enantiomers. AB - In A431 cells, depletion of cholesterol with methyl-beta-cyclodextrin induced an increase in both basal and epidermal growth factor (EGF)-stimulated EGF receptor phosphorylation. This increase in phosphorylation was site-specific, with significant increases occurring at Tyr845, Tyr992, and Tyr1173, but only minor changes at Tyr1045 and Tyr1068. The elevated level of receptor phosphorylation was associated with an increase in the intrinsic kinase activity of the EGF receptor kinase, possibly as a result of the cyclodextrin-induced enhancement of the phosphorylation of Tyr845, a site in the kinase activation loop known to be phosphorylated by pp60src. Cholesterol and its enantiomer (ent-cholesterol) were used to investigate the molecular basis for the modulation of EGF receptor function by cholesterol. Natural cholesterol (nat-cholesterol) was oxidized substantially more rapidly than ent-cholesterol by cholesterol oxidase, a protein that contains a specific binding site for the sterol. By contrast, the ability of nat- and ent-cholesterol to interact with sphingomyelins and phosphatidylcholine and to induce lipid condensation in a monolayer system was the same. These data suggest that, whereas cholesterol-protein interactions may be sensitive to the absolute configuration of the sterol, sterol-lipid interactions are not. nat- and ent-cholesterol were tested for their ability to physically reconstitute lipid rafts following depletion of cholesterol. nat- and ent-cholesterol reversed to the same extent the enhanced phosphorylation of the EGF receptor that occurred following removal of cholesterol. Furthermore, the enantiomers showed similar abilities to reconstitute lipid rafts in cyclodextrin-treated cells. These data suggest that cholesterol most likely affects EGF receptor function because of its physical effects on membrane properties, not through direct enantioselective interactions with the receptor. PMID- 14530281 TI - The conserved and non-conserved regions of Rpb4 are involved in multiple phenotypes in Saccharomyces cerevisiae. AB - Rpb4, the fourth largest subunit of RNA polymerase II in Saccharomyces cerevisiae, is required for many phenotypes, including growth at high and low temperatures, sporulation, pseudohyphal growth, activated transcription of a subset of genes, and efficient carbon and energy metabolism. We have used deletion analysis to delineate the domains of the protein involved in these multiple phenotypes. The scRpb4 protein is conserved at the N and C termini but possesses certain non-conserved regions in the central portion. Our deletion analysis and molecular modeling results show that the N- and C-terminal conserved regions of Rpb4 are involved in interaction with Rpb7, the Rpb4 interacting partner in the RNA polymerase II. We further show that the conserved N terminus is required for efficient activated transcription from the INO1 promoter but not the GAL10- or the HSE-containing promoters. The N terminus is not required for any of the stress responses tested: growth at high temperatures, sporulation, and pseudohyphal growth. The conserved C-terminal 23 amino acids are not required for the role of Rpb4 in the pseudohyphal growth phenotype but might play a role in other stress responses and activated transcription. From the deletion analysis of the non-conserved regions, we report that they influence phenotypes involving both the N and C termini (interaction with Rpb7 and transcription from the INO1 promoter) but not any of the stress-responsive phenotypes tested suggesting that they might be involved in maintaining the two conserved domains in an appropriate conformation for interaction with Rpb7 and other proteins. Taken together, our results allow us to assign phenotype-specific roles for the different conserved and non-conserved regions of Rpb4. PMID- 14530279 TI - A family of Ca2+-dependent activator proteins for secretion: comparative analysis of structure, expression, localization, and function. AB - Ca2+-dependent activator protein for secretion (CAPS) 1 is an essential cytosolic component of the protein machinery involved in large dense-core vesicle (LDCV) exocytosis and in the secretion of a subset of neurotransmitters. In the present study, we report the identification, cloning, and comparative characterization of a second mammalian CAPS isoform, CAPS2. The structure of CAPS2 and its function in LDCV exocytosis from PC12 cells are very similar to those of CAPS1. Both isoforms are strongly expressed in neuroendocrine cells and in the brain. In subcellular fractions of the brain, both CAPS isoforms are enriched in synaptic cytosol fractions and also present on vesicular fractions. In contrast to CAPS1, which is expressed almost exclusively in brain and neuroendocrine tissues, CAPS2 is also expressed in lung, liver, and testis. Within the brain, CAPS2 expression seems to be restricted to certain brain regions and cell populations, whereas CAPS1 expression is strong in all neurons. During development, CAPS2 expression is constant between embryonic day 10 and postnatal day 60, whereas CAPS1 expression is very low before birth and increases after postnatal day 0 to reach a plateau at postnatal day 21. Light microscopic data indicate that both CAPS isoforms are specifically enriched in synaptic terminals. Ultrastructural analyses show that CAPS1 is specifically localized to glutamatergic nerve terminals. We conclude that at the functional level, CAPS2 is largely redundant with CAPS1. Differences in the spatial and temporal expression patterns of the two CAPS isoforms most likely reflect as yet unidentified subtle functional differences required in particular cell types or during a particular developmental period. The abundance of CAPS proteins in synaptic terminals indicates that they may also be important for neuronal functions that are not exclusively related to LDCV exocytosis. PMID- 14530282 TI - Thermodynamic characterization of the binding of activator of G protein signaling 3 (AGS3) and peptides derived from AGS3 with G alpha i1. AB - Activator of G protein signaling 3 (AGS3) is a guanine nucleotide dissociation inhibitor (GDI) that contains four G protein regulatory (GPR) or GoLoco motifs in its C-terminal domain. The entire C-terminal domain (AGS3-C) as well as certain peptides corresponding to individual GPR motifs of AGS3 bound to G alpha i1 and inhibited the binding of GTP by stabilizing the GDP-bound conformation of G alpha i1. The stoichiometry, free energy, enthalpy, and dissociation constant for binding of AGS3-C to G alpha i1 were determined using isothermal titration calorimetry. AGS3-C possesses two apparent high affinity (Kd approximately 20 nm) and two apparent low affinity (Kd approximately 300 nm) binding sites for G alpha i1. Upon deletion of the C-terminal GPR motif from AGS3-C, the remaining sites were approximately equivalent with respect to their affinity (Kd approximately 400 nm) for G alpha i1. Peptides corresponding to each of the four GPR motifs of AGS3 (referred to as GPR1, GPR2, GPR3, and GPR4, respectively, going from N to C terminus) bound to G alpha i1 with Kd values in the range of 1-8 microm. Although GPR1, GPR2, and GPR4 inhibited the binding of the fluorescent GTP analog BODIPY FL-guanosine 5'-3-O-(thio)triphosphate to G alpha i1, GPR3 did not. However, addition of N- and C-terminal flanking residues to the GPR3 GoLoco core increased its affinity for G alpha i1 and conferred GDI activity similar to that of AGS3-C itself. Similar increases were observed for extended GPR2 and extended GPR1 peptides. Thus, while the tertiary structure of AGS3 may affect the affinity and activity of the GPR motifs contained within its sequence, residues outside of the GPR motifs strongly potentiate their binding and GDI activity toward G alpha i1 even though the amino acid sequences of these residues are not conserved among the GPR repeats. PMID- 14530283 TI - Expression profiling identifies genes that continue to respond to insulin in adipocytes made insulin-resistant by treatment with tumor necrosis factor-alpha. AB - We have employed microarray technology using RNA from normal 3T3-L1 adipocytes and from 3T3-L1 adipocytes made insulin-resistant by treatment with tumor necrosis factor-alpha to identify a new class of insulin-responsive genes. These genes continued to respond normally to insulin even though the adipocytes themselves were metabolically insulin-resistant, i.e. they displayed a significantly decreased rate of insulin-stimulated glucose uptake. Approximately 12,000 genes/expressed sequence tags (ESTs) were screened. Of these, 40 genes/ESTs were identified that became insulin-resistant as expected (e.g. Socs 3, junB, and matrix metalloproteinase-11). However, 61 genes/ESTs continued to respond normally to insulin. Although some of these genes were previously shown to be regulated by insulin (e.g. Glut-1 and beta3-adrenergic receptor), other novel insulin-sensitive genes were also identified (e.g. Egr-1, epiregulin, Fra 1, and ABCA1). Real-time reverse transcription-PCR analysis confirmed the expression patterns of several of the differentially expressed genes. One gene that remained insulin-sensitive in the insulin-resistant adipocytes is the transcription factor Egr-1. Using an antisense strategy, we show that tissue factor and macrophage colony-stimulating factor, two cardiovascular risk factors, are downstream EGR-1 target genes in the adipocyte. Taken together, these data support the hypothesis that some signaling pathways remain insulin-sensitive in metabolically insulin-resistant adipocytes. These pathways may promote abnormal gene expression in hyperinsulinemic states like obesity and type II diabetes and thus may contribute to pathologies associated with these conditions. PMID- 14530284 TI - Active PIKfyve associates with and promotes the membrane attachment of the late endosome-to-trans-Golgi network transport factor Rab9 effector p40. AB - PIKfyve, a kinase that displays specificity for phosphatidylinositol (PtdIns), PtdIns 3-phosphate (3-P), and proteins, is important in multivesicular body/late endocytic function. Enzymatically inactive PIKfyve mutants elicit enormous dilation of late endocytic structures, suggesting a role for PIKfyve in endosome to-trans-Golgi network (TGN) membrane retrieval. Here we report that p40, a Rab9 effector reported previously to bind Rab9-GTP and stimulate endosome-to-TGN transport, interacts with PIKfyve as determined by yeast two-hybrid assays, glutathione S-transferase (GST) pull-down assays, and co-immunoprecipitation in doubly transfected HEK293 cells. The interaction engages the PIKfyve chaperonin domain and four out of the six C-terminally positioned kelch repeats in p40. Differential centrifugation in a HEK293 cell line, stably expressing PIKfyveWT, showed the membrane-associated immunoreactive p40 co-sedimenting with PIKfyve in the high speed pellet (HSP) fraction. Remarkably, similar analysis in a HEK293 cell line stably expressing dominant-negative kinase-deficient PIKfyveK1831E demonstrated a marked depletion of p40 from the HSP fraction. GST-p40 failed to specifically associate with the PIKfyve lipid products PtdIns 5-P and PtdIns 3,5 P2 in a liposome binding assay but was found to be an in vitro substrate of the PIKfyve serine kinase activity. A band with the p40 electrophoretic mobility was found to react with a phosphoserine-specific antibody mainly in the PIKfyveWT containing fractions obtained by density gradient sedimentation of total membranes from PIKfyveWT-expressing HEK293 cells. Together these results identify the Rab9 effector p40 as a PIKfyve partner and suggest that p40-PIKfyve interaction and the subsequent PIKfyve-catalyzed p40 phosphorylation anchor p40 to discrete membranes facilitating late endosome-to-TGN transport. PMID- 14530285 TI - Induction of human methionine adenosyltransferase 2A expression by tumor necrosis factor alpha. Role of NF-kappa B and AP-1. AB - Two genes (MAT1A and MAT2A) encode for methionine adenosyltransferase (MAT), an essential cellular enzyme responsible for S-adenosylmethionine biosynthesis. MAT1A is expressed mostly in the liver, whereas MAT2A is widely distributed. We showed a switch from MAT1A to MAT2A expression in human hepatocellular carcinoma (HCC), which facilitates cancer cell growth. Using DNase I footprinting analysis, we previously identified a region in the MAT2A promoter protected from DNase I digestion in HCC. This region contains NF-kappa B and AP-1 elements, and the present study examined whether they regulate MAT2A promoter activity. We found nuclear binding of NF-kappa B and AP-1 to the MAT2A promoter increased in HCC. Tumor necrosis factor alpha (TNFalpha), which activates both NF-kappa B and AP-1, increased MAT2A expression in a dose- and time-dependent manner, binding of both NF-kappa B and AP-1 to the MAT2A promoter and MAT2A promoter activity, with the latter effect blocked by site-directed mutagenesis of the NF-kappa B and AP-1 binding sites. Blocking NF-kappa B with I kappa B super-repressor or AP-1 with dominant-negative c-Jun led to decreased basal MAT2A expression and prevented the TNF alpha-induced increase in MAT2A expression. Although blocking NF-kappa B had no influence on the ability of TNF alpha to increase AP-1 nuclear binding, blocking AP-1 with dominant-negative c-Jun prevented the TNF alpha-mediated increase in NF-kappa B binding. In conclusion, both NF-kappa B and AP-1 are required for basal MAT2A expression in HepG2 cells and mediate the increase in MAT2A expression in response to TNF alpha treatment. Increased trans-activation of these two sites also contributes to MAT2A up-regulation in HCC. PMID- 14530286 TI - Involvement of nectin-activated Cdc42 small G protein in organization of adherens and tight junctions in Madin-Darby canine kidney cells. AB - Nectins, Ca2+-independent immunoglobulin-like cell-cell adhesion molecules, trans interact and form cell-cell adhesion, which increases the velocities of the formation of the E-cadherin-based adherens junctions (AJs) and the claudin-based tight junctions (TJs) in Madin-Darby canine kidney (MDCK) cells. The trans interactions of nectins furthermore induce activation of Cdc42 and Rac small G proteins, but the roles of these small G proteins activated in this way remain unknown. We examined here the role and the mode of action of Cdc42 in the organization of AJs and TJs in MDCK cells. We first made the NWASP-Cdc42 and Rac interactive binding (CRIB) domain, an inhibitor of activated Cdc42, fused to the Ki-Ras CAAX motif (NWASP-CRIB-CAAX; where A is aliphatic amino acid), which was targeted to the cell-cell adhesion sites. We then found that overexpression of NWASP-CRIB-CAAX reduced the velocities of the formation of AJs and TJs. Conversely, overexpression of a constitutively active mutant of Cdc42 (V12Cdc42) increased their velocities, and the inhibitory effect of NWASP-CRIB-CAAX was suppressed by co-expression with V12Cdc42. The inhibitory effect of NWASP-CRIB CAAX on the formation of AJs and TJs was suppressed by co-expression of nectin-1 of which trans-interaction activated endogenous Cdc42. Moreover, the formation of the claudin-based TJs required a greater amount of activated Cdc42 than that of the E-cadherin-based AJs. These results indicate that the Cdc42 activated by the trans-interactions of nectins is involved in the organization of AJs and TJs in different mechanisms in MDCK cells. PMID- 14530287 TI - The retinoic acid-responsive proline-rich protein is identified in promyeloleukemic HL-60 cells. AB - To identify new genes that retinoic acid activates, we employed an mRNA differential display technique and screened for genes that are differentially expressed in promyeloleukemic HL-60 cells incubated in the presence of all-trans retinoic acid (ATRA) compared with the absence of ATRA. We cloned the coding region of a retinoic acid-induced gene from a human thymus library, which was the mRNA encoding the 666-amino acid human homologue of mouse proline-rich protein 76. We have designated it RARP1 (retinoic acid response proline-rich protein 1). Transcription of an approximately 2.4-kbp mRNA occurred mainly in organs with immune functions, such as thymus, spleen, and peripheral leukocytes. Cycloheximide blocked the ATRA-induced expression. In megakaryocyte-like human erythroleukemia HEL cells, the amount of RARP1 mRNA was high, but it was low in human T-lymphoblastoid Jurkat cells. A specific antibody against RARP1 recognized a 110-kDa protein, which accumulates after incubation of HL-60 cells with ATRA. In immunohistochemical experiments, strong RARP1 staining was observed in the megakaryocytes of bone marrow and spleen, and heterogeneous stain was seen in thymus. Transcriptional studies showed that RARP1 expression impaired the transactivation through activator protein1 and serum response-element in all cell lines we checked, whereas it did not affect the transactivation through cAMP response element in the same cell lines. Further analysis demonstrated that proline-rich regions of RARP1 are the functional regions regulated for suppression of activator protein1 transactivation. These data suggest that ATRA inducible RARP1 selectively affects signal transduction and may contribute to myeloid and megakaryocytic differentiation. PMID- 14530288 TI - Exploitation of a chemical nuclease to investigate the location and orientation of the Escherichia coli RNA polymerase alpha subunit C-terminal domains at simple promoters that are activated by cyclic AMP receptor protein. AB - The C-terminal domain of the alpha subunit (alphaCTD) of bacterial RNA polymerase plays an important role in promoter recognition. It is known that alphaCTD binds to the DNA minor groove at different locations at different promoters via a surface-exposed determinant, the 265 determinant. Here we describe experiments that permit us to determine the location and orientation of binding of alphaCTD at any promoter. In these experiments, a DNA cleavage reagent is attached to specific locations on opposite faces of the RNA polymerase alpha subunit. After incorporation of the tagged alpha subunits into holo-RNA polymerase, patterns of DNA cleavage due to the reagent are determined in open complexes. The locations of DNA cleavage due to the reagent attached at different positions allow the position and orientation of alphaCTD to be deduced. Here we present data from experiments with simple Escherichia coli promoters that are activated by the cyclic AMP receptor protein. PMID- 14530289 TI - Inhibition of fructose-1,6-bisphosphatase by a new class of allosteric effectors. AB - A highly constrained pseudo-tetrapeptide (OC252-324) further defines a new allosteric binding site located near the center of fructose-1,6-bisphosphatase. In a crystal structure, pairs of inhibitory molecules bind to opposite faces of the enzyme tetramer. Each ligand molecule is in contact with three of four subunits of the tetramer, hydrogen bonding with the side chain of Asp187 and the backbone carbonyl of residue 71, and electrostatically interacting with the backbone carbonyl of residue 51. The ligated complex adopts a quaternary structure between the canonical R- and T-states of fructose-1,6-bisphosphatase, and yet a dynamic loop essential for catalysis (residues 52-72) is in a conformation identical to that of the T-state enzyme. Inhibition by the pseudo tetrapeptide is cooperative (Hill coefficient of 2), synergistic with both AMP and fructose 2,6-bisphosphate, noncompetitive with respect to Mg2+, and uncompetitive with respect to fructose 1,6-bisphosphate. The ligand dramatically lowers the concentration at which substrate inhibition dominates the kinetics of fructose-1,6-bisphosphatase. Elevated substrate concentrations employed in kinetic screens may have facilitated the discovery of this uncompetitive inhibitor. Moreover, the inhibitor could mimic an unknown natural effector of fructose-1,6-bisphosphatase, as it interacts strongly with a conserved residue of undetermined functional significance. PMID- 14530290 TI - Unzipping the role of myosin light chain phosphatase in smooth muscle cell relaxation. AB - Recently, it has been hypothesized that myosin light chain (MLC) phosphatase is activated by cGMP-dependent protein kinase (PKG) via a leucine zipper-leucine zipper (LZ-LZ) interaction through the C-terminal LZ in the myosin-binding subunit (MBS) of MLC phosphatase and the N-terminal LZ of PKG (Surks, H. K., Mochizuki, N., Kasai, Y., Georgescu, S. P., Tang, K. M., Ito, M., Lincoln, T. M., and Mendelsohn, M. E. (1999) Science 286, 1583-1587). Alternative splicing of a 3'-exon produces a LZ+ or LZ- MBS, and the sensitivity to cGMP-mediated smooth muscle relaxation correlates with the relative expression of LZ+/LZ- MBS isoforms (Khatri, J. J., Joyce, K. M., Brozovich, F. V., and Fisher, S. A. (2001) J. Biol. Chem. 276, 37250 -37257). In the present study, we determined the effect of LZ+/LZ- MBS isoforms on cGMP-induced MLC20 dephosphorylation. Four avian smooth muscle MBS-recombinant adenoviruses were prepared and transfected into cultured embryonic chicken gizzard smooth muscle cells. The expressed exogenous MBS isoforms were shown to replace the endogenous isoform in the MLC phosphatase holoenzyme. The interaction of type I PKG (PKGI) with the MBS did not depend on the presence of cGMP or the MBS LZ. However, direct activation of PKGI by 8-bromo cGMP produced a dose-dependent decrease in MLC20 phosphorylation (p<0.05) only in smooth muscle cells expressing a LZ+ MBS. These results suggest that the activation of MLC phosphatase by PKGI requires a LZ+ MBS, but the binding of PKGI to the MBS is not mediated by a LZ-LZ interaction. Thus, the relative expression of LZ+/LZ- MBS isoforms could explain differences in tissue sensitivity to NO mediated vasodilatation. PMID- 14530291 TI - A DNA pairing-enhanced conformation of bacterial RecA proteins. AB - The RecA proteins of Escherichia coli (Ec) and Deinococcus radiodurans (Dr) both promote a DNA strand exchange reaction involving two duplex DNAs. The four-strand exchange reaction promoted by the DrRecA protein is similar to that promoted by EcRecA, except that key parts of the reaction are inhibited by Ec single-stranded DNA-binding protein (SSB). In the absence of SSB, the initiation of strand exchange is greatly enhanced by dsDNA-ssDNA junctions at the ends of DNA gaps. This same trend is seen with the EcRecA protein. The results lead to an expansion of published hypotheses for the pathway for RecA-mediated DNA pairing, in which the slow first order step (observed in several studies) involves a structural transition to a state we designate P. The P state is identical to the state found when RecA is bound to double-stranded (ds) DNA. The structural state present when the RecA protein is bound to single-stranded (ss) DNA is designated A. The DNA pairing model in turn facilitates an articulation of three additional conclusions arising from the present work. 1) When a segment of a RecA filament bound to ssDNA is forced into the P state (as RecA bound to the ssDNA immediately adjacent to dsDNA-ssDNA junction), the segment becomes "pairing enhanced." 2) The unusual DNA pairing properties of the D. radiodurans RecA protein can be explained by postulating this protein has a more stringent requirement to initiate DNA strand exchange from the P state. 3) RecA filaments bound to dsDNA (P state) have directly observable structural changes relative to RecA filaments bound to ssDNA (A state), involving the C-terminal domain. PMID- 14530292 TI - Genetic determinants of nonmodulating hypertension. AB - We sought to determine whether genes of the renin-angiotensin-aldosterone system can predict the nonmodulating intermediate phenotype in essential hypertension. Aldosterone responses to angiotensin II were assessed in 298 subjects with hypertension. Subjects were genotyped at the angiotensinogen M235T, angiotensin converting enzyme I/D, aldosterone synthase C-344 T, renin, angiotensin II type 1 receptor, and adducin loci. The data were analyzed by Student t test, ANOVA, stepwise linear regression and general linear model or GENMOD regression techniques, and chi2 analysis odds ratios (ORs). Aldosterone response varied by genotype for angiotensin and aldosterone synthase but not for the other loci. The combination of angiotensinogen 235 TT and angiotensin-converting enzyme DD showed further reduction (P=0.0377) when compared with angiotensinogen 235 TT alone, an example of genetic epistasis. When the subject was required also to possess the CYP11B2 -344 TT genotype, there was a further substantial reduction. Of these 3 loci, only angiotensinogen 235 TT significantly increased the OR of predicting the nonmodulating hypertensive phenotype (OR, 2.00; 95% confidence interval, 1.152 to 3.51). However, when angiotensin-converting enzyme DD was combined with angiotensinogen 235 TT, the OR nearly doubled to 3.74, with a further increase to 5.36-fold when the subject possessed all 3 genotypes. Thus, the angiotensinogen, angiotensin-converting enzyme, and aldosterone synthase genotypes identified individuals with the nonmodulating phenotype with an increasing degree of fidelity. For this subclass of essential hypertension, it is likely that genotyping can be substituted for complex phenotyping for therapeutic and preventive decision making. PMID- 14530293 TI - Drosophila Echinoid is an antagonist of Egfr signalling, but is not a member of the L1-type family of cell adhesion molecules. PMID- 14530294 TI - Wnts and Hedgehogs: lipid-modified proteins and similarities in signaling mechanisms at the cell surface. AB - This review compares the signaling mechanisms of the Wnt and the Hedgehog proteins. Although Wnts and Hedgehogs are unrelated proteins, they are both modified by lipids, possibly through the action of enzymes that are related to each other. At the surface of target cells, the reception of Wnt and Hedgehog signals is regulated by several molecules, some of which, in particular the Frizzled and Smoothened receptors, are related to each other. Several other aspects of Wnt and Hedgehog transport and signaling are discussed, as well as the possible origin of these pathways. PMID- 14530295 TI - A crucial role for Fgfr2-IIIb signalling in epidermal development and hair follicle patterning. AB - To understand the role Fgf signalling in skin and hair follicle development, we analysed the phenotype of mice deficient for Fgfr2-IIIb and its main ligand Fgf10. These studies showed that the severe epidermal hypoplasia found in mice null for Fgfr2-IIIb is caused by a lack of the basal cell proliferation that normally results in a stratified epidermis. Although at term the epidermis of Fgfr2-IIIb null mice is only two to three cells thick, it expresses the classical markers of epidermal differentiation and establishes a functional barrier. Mice deficient for Fgf10 display a similar but less severe epidermal hypoplasia. By contrast, Fgfr2-IIIb-/-, but not Fgf10-/-, mice produced significantly fewer hair follicles, and their follicles were developmentally retarded. Following transplantation onto nude mice, grafts of Fgfr2-IIIb-/- skin showed impaired hair formation, with a decrease in hair density and the production of abnormal pelage hairs. Expression of Lef1, Shh and Bmp4 in the developing hair follicles of Fgfr2 IIIb-/- mice was similar to wild type. These results suggest that Fgf signalling positively regulates the number of keratinocytes needed to form a normal stratified epidermis and to initiate hair placode formation. In addition, Fgf signals are required for the growth and patterning of pelage hairs. PMID- 14530296 TI - The maternally expressed zebrafish T-box gene eomesodermin regulates organizer formation. AB - Early embryonic development in many organisms relies upon maternal molecules deposited into the egg prior to fertilization. We have cloned and characterized a maternal T-box gene in the zebrafish, eomesodermin (eomes). During oogenesis, the eomes transcript becomes localized to the cortex of the oocyte. After fertilization during early cleavage stages, eomes is expressed in a vegetal to animal gradient in the embryo, whereas Eomesodermin protein (Eom) is distributed cytoplasmically throughout the blastoderm. Strikingly, following midblastula transition, nuclear-localized Eomesodermin is detected on the dorsal side of the embryo only. Overexpression of eomes results in Nodal-dependent and nieuwkoid/dharma (nwk/dhm) independent ectopic expression of the organizer markers goosecoid (gsc), chordin (chd) and floating head (flh) and in the formation of secondary axes. The same phenotypes are observed when a VP16 activator construct is injected into early embryos, indicating that eomes acts as a transcriptional activator. In addition, a dominant-negative construct and antisense morpholino oligonucleotides led to a reduction in gsc and flh expression. Together these data indicate that eomes plays a role in specifying the organizer. PMID- 14530297 TI - Trophoblasts acquire a chemokine receptor, CCR1, as they differentiate towards invasive phenotype. AB - At the human feto-maternal interface, trophoblasts differentiate towards extravillous trophoblasts (EVTs) and form the cell column. EVTs acquire invasive activity in the distal part of the cell column and begin to migrate into the maternal tissue. We previously reported that dipeptidyl peptidase IV (DPPIV) is expressed on EVTs in the proximal part of cell column and is involved in the inhibition of their migration. Because DPPIV has been shown to degrade several chemokines, we examined possible roles of chemokines in EVT migration. Immunohistochemistry demonstrated that C-C chemokine receptor 1 (CCR1) was hardly detected on cytotrophoblasts and syncytiotrophoblast but was expressed on EVTs in the cell column. In vitro, CCR1 protein was also present on the surface of EVTs that grew out from chorionic villous explants cultured under 20% O2. Chemokines that can bind to CCR1 (CCR1 ligands), such as regulated on activation, normal T cell expressed and secreted (RANTES) and macrophage inflammatory protein-1alpha (MIP-1alpha), were confirmed in the decidual tissues by RT-PCR and immunohistochemistry. These CCR1 ligands promoted the migration of the EVTs that were isolated from the explant cultures in vitro. These results indicate that CCR1 is expressed on trophoblasts as they differentiate to EVTs and that CCR1 ligands produced from the decidual tissue induce EVT migration. By contrast, CCR1 was scarcely expressed on EVTs that grew out from villous explants cultured in 1% O2, indicating that a relatively high oxygenic environment is needed to induce CCR1 expression. Moreover, CCR1 expression on the isolated EVTs was significantly reduced in the presence of decidua-conditioned medium. Such regulation of CCR1 by surrounding oxygenic and decidual environments supports a close correlation between EVT invasion and their expression of CCR1. This study demonstrates that trophoblasts acquire CCR1 as they differentiate to an invasive phenotype at the villus-anchoring sites and indicates a novel role for the chemokine-CCR1 system in the initial step of trophoblastic invasion towards the maternal tissue. PMID- 14530298 TI - Functional network integration of embryonic stem cell-derived astrocytes in hippocampal slice cultures. AB - Embryonic stem (ES) cells provide attractive prospects for neural transplantation. So far, grafting strategies in the CNS have focused mainly on neuronal replacement. Employing a slice culture model, we found that ES cell derived glial precursors (ESGPs) possess a remarkable capacity to integrate into the host glial network. Following deposition on the surface of hippocampal slices, ESGPs actively migrate into the recipient tissue and establish extensive cell-cell contacts with recipient glia. Gap junction-mediated coupling between donor and host astrocytes permits widespread delivery of dye from single donor cells. During maturation, engrafted donor cells display morphological, immunochemical and electrophysiological properties that are characteristic of differentiating native glia. Our findings provide the first evidence of functional integration of grafted astrocytes, and depict glial network integration as a potential route for widespread transcellular delivery of small molecules to the CNS. PMID- 14530299 TI - Control of dendritic development by the Drosophila fragile X-related gene involves the small GTPase Rac1. AB - Fragile X syndrome is caused by loss-of-function mutations in the fragile X mental retardation 1 gene. How these mutations affect neuronal development and function remains largely elusive. We generated specific point mutations or small deletions in the Drosophila fragile X-related (Fmr1) gene and examined the roles of Fmr1 in dendritic development of dendritic arborization (DA) neurons in Drosophila larvae. We found that Fmr1 could be detected in the cell bodies and proximal dendrites of DA neurons and that Fmr1 loss-of-function mutations increased the number of higher-order dendritic branches. Conversely, overexpression of Fmr1 in DA neurons dramatically decreased dendritic branching. In dissecting the mechanisms underlying Fmr1 function in dendrite development, we found that the mRNA encoding small GTPase Rac1 was present in the Fmr1-messenger ribonucleoprotein complexes in vivo. Mosaic analysis with a repressor cell marker (MARCM) and overexpression studies revealed that Rac1 has a cell-autonomous function in promoting dendritic branching of DA neurons. Furthermore, Fmr1 and Rac1 genetically interact with each other in controlling the formation of fine dendritic branches. These findings demonstrate that Fmr1 affects dendritic development and that Rac1 is partially responsible for mediating this effect. PMID- 14530301 TI - ENU large-scale mutagenesis and quantitative trait linkage (QTL) analysis in mice: novel technologies for searching polygenetic determinants of craniofacial abnormalities. AB - Discrepancies in size and shape of the jaws are the underlying etiology in many orthodontic and orthognathic surgery patients. Genetic factors combined with environmental interactions have been postulated to play a causal or contributory role in these craniofacial abnormalities. Along with the soon-to-be-available complete human and mouse genomic sequence data, mouse mutants have become a valuable tool in the functional mapping of genes involved in the development of human maxillofacial dysmorphologies. We review two powerful methods in such efforts: N-ethyl-N-nitrosourea (ENU) large-scale mutagenesis and quantitative trait linkage (QTL) analysis. The former aims at producing a plethora of novel variants of particular trait(s), and ultimately mapping the point mutations responsible for the appearance of these new traits. In contrast, the latter applies intensive breeding and mapping techniques to identify multiple loci (and, subsequently, genes) contributing to the phenotypic difference between the tested strains. A prerequisite for either approach to studying variations in the traits of interest is the application of effective mouse cephalometric phenotype analysis and rapid DNA mapping techniques. These approaches will produce a wealth of new data on critical genes that influence the size and shape of the human face. PMID- 14530302 TI - Molecular genetic analysis of the virulence of oral bacterial pathogens: an historical perspective. AB - This review will focus on the impact of molecular genetic approaches on elucidating the bacterial etiology of oral diseases from an historical perspective. Relevant results from the pre- and post-recombinant DNA periods will be highlighted, including the roles of gene cloning, mutagenesis, and nucleotide sequencing in this area of research. Finally, the impact of whole-genome sequencing on deciphering the virulence mechanisms of oral pathogens, along with new approaches to control these organisms, will be discussed. PMID- 14530303 TI - Development of HPV vaccines for HPV-associated head and neck squamous cell carcinoma. AB - High-risk genotypes of the human papillomavirus (HPV), particularly HPV type 16, are found in a distinct subset of head and neck squamous cell carcinomas (HNSCC). Thus, these HPV-associated HNSCC may be prevented or treated by vaccines designed to induce appropriate HPV virus-specific immune responses. Infection by HPV may be prevented by neutralizing antibodies specific for the viral capsid proteins. In clinical trials, vaccines comprised of HPV virus-like particles (VLPs) have shown great promise as prophylactic HPV vaccines. However, given that capsid proteins are not expressed at detectable levels by infected basal keratinocytes, vaccines with therapeutic potential must target other non-structural viral antigens. Two HPV oncogenic proteins, E6 and E7, are important in the induction and maintenance of cellular transformation and are co-expressed in the majority of HPV-containing carcinomas. Therefore, therapeutic vaccines targeting these proteins may have potential to control HPV-associated malignancies. Various candidate therapeutic HPV vaccines are currently being tested whereby E6 and/or E7 is administered in live vectors, in peptides or protein, in nucleic acid form, as components of chimeric VLPs, or in cell-based vaccines. Encouraging results from experimental vaccination systems in animal models have led to several prophylactic and therapeutic vaccine clinical trials. Should they fulfill their promise, these vaccines may prevent HPV infection or control its potentially life threatening consequences in humans. PMID- 14530304 TI - The molecular biology of mucosal field cancerization of the head and neck. AB - Field cancerization was first described in 1953 as histologically altered epithelium surrounding tumor samples taken from the upper aerodigestive tract. Since then, the term has been used to describe multiple patches of pre-malignant disease, a higher-than-expected prevalence of multiple local second primary tumors, and the presence of synchronous distant tumors within the upper aerodigestive tract. Molecular techniques such as karyotype analysis, microsatellite analysis, p53 mutation screening, and X-chromosome inactivation studies have further refined the relationship among these lesions. While there are differences in the techniques used to identify the clonal origins of the lesions, these studies indicate that there is often lateral clonal spread of pre malignant or malignant disease, and a significant portion of local second primary tumors are in fact genetically related. Distant second primary tumors found in the esophagus are often not related to concurrent head and neck cancer, whereas synchronous squamous lung tumors with a head and neck primary are often, in fact, metastases, rather than independently arising malignancies. These observations help to explain the high incidence of recurrent disease, despite excision or other therapy--pre-malignant or malignant clones often have the ability to migrate and persist outside of the field of treatment. Therefore, alternative means of prevention or therapy that can affect the entire head and neck region may be of benefit to such patients. Future studies will further refine the relationship among these lesions and perhaps identify key molecular alterations to be used as targets for gene therapy. PMID- 14530305 TI - Molecular recognition at the protein-hydroxyapatite interface. AB - Proteins found in mineralized tissues act as nature's crystal engineers, where they play a key role in promoting or inhibiting the growth of minerals such as hydroxyapatite (bones/teeth) and calcium oxalate (kidney stones). Despite their importance in hard-tissue formation and remodeling, and in pathological processes such as stone formation and arterial calcification, there is little known of the protein structure-function relationships that govern hard-tissue engineering. Here we review early studies that have utilized solid-state NMR (ssNMR) techniques to provide in situ secondary-structure determination of statherin and statherin peptides on their biologically relevant hydroxyapatite (HAP) surfaces. In addition to direct structural study, molecular dynamics studies have provided considerable insight into the protein-binding footprint on hydroxyapatite. The molecular insight provided by these studies has also led to the design of biomimetic fusion peptides that utilize nature's crystal-recognition mechanism to display accessible and dynamic bioactive sequences from the HAP surface. These peptides selectively engage adhesion receptors and direct specific outside-in signaling pathway activation in osteoblast-like cells. PMID- 14530306 TI - Paracoccidioidomycosis of the mouth: an emerging deep mycosis. AB - Oral fungal infections (mycoses) have come into particular prominence since the advent of infection with Human Immunodeficiency Virus (HIV), and recognition of the Acquired Immune Deficiency Syndrome (AIDS), as well as the phenomenal increase in world travel with increased exposure to infections endemic in the tropics. Paracoccidioidomycosis is a rare mycosis worldwide but common in Brazil and some other areas in Latin America. It can be life-threatening and can manifest with a spectrum of clinical presentations, including frequent oral lesions. This paper reviews the more recent information on Paracoccidioidomycosis, emphasizing those areas most relevant in dental science. PMID- 14530307 TI - Cutting edge: cyclooxygenase-2 activation suppresses Th1 polarization in response to Helicobacter pylori. AB - Helicobacter pylori infection causes a Th1-driven mucosal immune response. Cyclooxygenase (COX)-2 is up-regulated in lamina propria mononuclear cells in H. pylori gastritis. Because COX-2 can modulate Th1/Th2 balance, we determined whether H. pylori activates COX-2 in human PBMCs, and the effect on cytokine and proliferative responses. There was significant up-regulation of COX-2 mRNA and PGE(2) release in response to H. pylori preparations. Addition of COX-2 inhibitors or an anti-PGE(2) Ab resulted in a marked increase in H. pylori stimulated IL-12 and IFN-gamma production, and a decrease in IL-10 levels. Addition of PGE(2) or cAMP, the second messenger activated by PGE(2), had the opposite effect. Similarly, stimulated cell proliferation was increased by COX-2 inhibitors or anti-PGE(2) Ab, and was decreased by PGE(2). Our findings indicate that COX-2 has an immunosuppressive role in H. pylori gastritis, which may protect the mucosa from severe injury, but may also contribute to the persistence of the infection. PMID- 14530309 TI - Different dynamics of CD4+ and CD8+ T cell responses during and after acute lymphocytic choriomeningitis virus infection. AB - We fit a mathematical model to data characterizing the primary cellular immune response to lymphocytic choriomeningitis virus. The data enumerate the specific CD8(+) T cell response to six MHC class I-restricted epitopes and the specific CD4(+) T cell responses to two MHC class II-restricted epitopes. The peak of the response occurs around day 8 for CD8(+) T cells and around day 9 for CD4(+) T cells. By fitting a model to the data, we characterize the kinetic differences between CD4(+) and CD8(+) T cell responses and among the immunodominant and subdominant responses to the various epitopes. CD8(+) T cell responses have faster kinetics in almost every aspect of the response. For CD8(+) and CD4(+) T cells, the doubling time during the initial expansion phase is 8 and 11 h, respectively. The half-life during the contraction phase following the peak of the response is 41 h and 3 days, respectively. CD4(+) responses are even slower because their contraction phase appears to be biphasic, approaching a 35-day half life 8 days after the peak of the response. The half-life during the memory phase is 500 days for the CD4(+) T cell responses and appears to be lifelong for the six CD8(+) T cell responses. Comparing the responses between the various epitopes, we find that immunodominant responses have an earlier and/or larger recruitment of precursors cells before the expansion phase and/or have a faster proliferation rate during the expansion phase. PMID- 14530308 TI - Specific inhibition of Stat5a/b promotes apoptosis of IL-2-responsive primary and tumor-derived lymphoid cells. AB - Stat5a/b exhibits 96% homology and are required for normal immune function. The present studies examined Stat5a/b function in lymphoid cells by specific and simultaneous disruption of both proteins using novel phosphorothioate-2'-O methoxyethyl antisense oligodeoxynucleotides (asODN). Efficient delivery was confirmed by the presence of fluorescent TAMRA-labeled ODN in >or=55 and 95% in human primary and tumor cell lines, respectively. Acute asODN administration reduced levels of Stat5a (90%) in 6 h, whereas Stat5b required nearly 48 h to attain the same inhibition, suggesting that the apparent turnover rate for Stat5a was 8-fold higher than that for Stat5b. Expression of the closely related Stat3 protein was unchanged after asODN treatment, however. Molecular ablation of Stat5a/b promoted apoptotic cell death in a significant population of primary PHA activated T cells (72%) and lymphoid tumor cell line (e.g., YT; 74%) within 24 h, as assessed by 1) visualization of karyolytic nuclear degeneration and other generalized cytoarchitectural alterations, 2) enzymatic detection of TdT-positive DNA degradation, and 3) automated cytometric detection of annexin V translocation. Contrary to findings from Stat5a/b-null mice, cell cycle progression did not appear to be significantly affected. Interestingly, IL-2 insensitive and unprimed T cells and Jurkat cells remained mostly unaffected. Finally, evidence is provided that the cytotoxicity associated with Stat5a/b ablation may derive from activation of caspase-8, an initiator protease that contributes to apoptotic cell commitment. We propose that in lymphoid cells competent to activate Stat5a and Stat5b, both proteins preferentially mediate an antiapoptotic survival influence. PMID- 14530310 TI - Prostaglandin D2 inhibits airway dendritic cell migration and function in steady state conditions by selective activation of the D prostanoid receptor 1. AB - PGD(2) is the major mediator released by mast cells during allergic responses, and it acts through two different receptors, the D prostanoid receptor 1 (DP1) and DP2, also known as CRTH2. Recently, it has been shown that PGD(2) inhibits the migration of epidermal Langerhans cells to the skin draining lymph nodes (LNs) and affects the subsequent cutaneous inflammatory reaction. However, the role of PGD(2) in the pulmonary immune response remains unclear. Here, we show that the intratracheal instillation of FITC-OVA together with PGD(2) inhibits the migration of FITC(+) lung DC to draining LNs. This process is mimicked by the DP1 agonist BW245C, but not by the DP2 agonist DK-PGD(2). The ligation of DP1 inhibits the migration of FITC-OVA(+) DCs only temporarily, but still inhibits the proliferation of adoptively transferred, OVA-specific, CFSE-labeled, naive T cells in draining LNs. These T cells produced lower amounts of the T cell cytokines IL-4, IL-10, and IFN-gamma compared with T cells from mice that received FITC-OVA alone. Taken together, our data suggest that the activation of DP receptor by PGD(2) may represent a pathway to control airway DC migration and to limit the activation of T cells in the LNs under steady state conditions, possibly contributing to homeostasis in the lung. PMID- 14530311 TI - Intratumor CpG-oligodeoxynucleotide injection induces protective antitumor T cell immunity. AB - Tumor cells are typically poorly immunogenic. The same mechanisms that evolved to avoid the induction of immune responses against self tissues, and, hence, autoimmune disease, also have to be overcome for immune therapy of cancer. Toll like receptor-activating microbial products such as CpG motif containing DNA are among the primary stimuli that the immune system uses to distinguish between infectious nonself (that is to be attacked) and noninfectious self (that must not be attacked). We tested in a murine RMA lymphoma/C57BL/6 model whether providing the infectious nonself context in a tumor-by injecting CpG-oligodeoxynucleotides directly into the tumor-would elicit a protective antitumor response. Complete remission of established solid tumors was achieved in immune competent mice, but not in T cell/B cell-deficient RAG-1 knockout mice. Intratumor injection of CpG oligodeoxynucleotides was shown to induce a tumor-specific CD4(+) and CD8(+) T cell response of the type 1 effector class, and T cells adoptively transferred the protection to RAG-1 knockout mice. The data show that intratumor injection of CpG-oligodeoxynucleotides is a promising strategy for rendering tumors immunogenic. PMID- 14530312 TI - Mediation of enhanced transcription of the IL-10 gene in T cells, upon contact with human glioma cells, by Fas signaling through a protein kinase A-independent pathway. AB - Elevated expression of IL-10 has been frequently observed in tumor tissues and tumor-infiltrating cells. We show herein that transcription of the IL-10 gene in primary peripheral T cells and T cell lines is up-regulated upon contact with glioma cells without an induction of apoptosis in those T cells. Glioma associated IL-10 induction was suppressed by interrupting the engagement of Fas and its ligand (Fas-L) with the antagonistic Ab, ZB4, by reducing Fas-L expression of glioma cells using the Fas-L-specific ribozyme, or by preventing cell-to-cell contact in a Transwell culture setting. Cross-linking of Fas with the agonistic Ab, CH-11, triggered apoptosis and enhanced the expression of IL-10 in Jurkat cells at the transcriptional and translational levels. Inhibiting caspase activities by caspase inhibitors, Z-VAD (Z-Val-Ala-Asp(Ome) fluoromethylketone) and Z-IETD (Z-Ile-Glu(Ome)-Thr(Ome)-Asp(Ome) fluoromethylketone), abolished this IL-10 induction in Jurkat cells. Intracellular staining detected IL-10 proteins in Fas-cross-linked Jurkat cells and in PHA-activated T cells. However, few IL-10 proteins were detectable in Jurkat cells cocultured with glioma cells, indicating a requirement of other factors for IL-10 production. Direct activation of protein kinase A (PKA) by forskolin elevated the transcription of IL-10 in Jurkat cells. However, KT5720, a selective PKA inhibitor, reduced neither anti-Fas-triggered nor glioma-associated IL-10 expression. Phosphorylation of cAMP response element binding protein and activating transcription factor-1 in Jurkat cells was not affected by coculturing with glioma cells or by anti-Fas treatment, further suggesting a PKA-independent pathway. In summary, our results demonstrate nonlethal cross-talk between tumor and immune cells leading to IL-10 dysregulation in T cells, which might contribute to Fas-L(+) tumor-associated immunosuppression. PMID- 14530313 TI - Functional effects of TNF-alpha on a human follicular dendritic cell line: persistent NF-kappa B activation and sensitization for Fas-mediated apoptosis. AB - Follicular dendritic cells (FDC) play crucial roles in germinal center (GC) formation and differentiation of GC B cells. FDC functions are influenced by cytokines produced in the GC. Among the GC cytokines, TNF is known to be essential for the formation and maintenance of the FDC network in the GC. We found that TNF is a mitogenic growth factor to an established FDC-like cell line, HK cells. Differing from most cell types which become desensitized to TNF action, HK cells exhibited persistent TNF signaling, as demonstrated by prolonged and biphasic NF-kappaB activation even after 3 days of TNF treatment. As a result, antiapoptotic genes including TNFR-associated factors 1 and 2, and cellular inhibitor of apoptosis proteins 1 and 2 were persistently induced by TNF, leading to the protection against TNF-mediated cell death. However, TNF pretreatment enhanced Fas-mediated apoptosis by up-regulating surface Fas expression in an NF kappaB-dependent pathway. During the GC responses, proliferation followed by FDC death has not been documented. However, our in vitro results suggest that FDCs proliferate in response to TNF, and die by Fas-mediated apoptosis whose susceptibility is enhanced by TNF, representing a mode of action for TNF in the maintenance of FDC networks by regulating the survival or death of FDC. PMID- 14530314 TI - Regulation of developing B cell survival by RelA-containing NF-kappa B complexes. AB - Mice deficient in the RelA (p65) subunit of NF-kappaB die during embryonic development. Fetal liver (FL) hemopoietic precursors from these mice were used to generate RelA-deficient lymphocytes by adoptive transfer into lethally irradiated mature lymphocyte-deficient recombination-activating gene-1(-/-) mice. Strikingly, RelA(-/-) lymphocyte generation was greatly diminished compared with that of RelA(+/+) lymphocytes. The most dramatic reduction was noticed in the numbers of developing B cells, which were considerably increased when RelA(-/-) FL cells that were also TNFR1 deficient were used. The role of RelA was further investigated in FL-derived developing B cells in vitro. Our results show that RelA is a major component of constitutive and TNF-alpha-induced kappaB site binding activity in developing B cells, and provide evidence for a direct role of TNF-alpha in killing RelA(-/-) B cells. The absence of RelA significantly reduced mRNA expression of the antiapoptotic genes cellular FLICE-inhibitory protein and Bcl-2. Retroviral transduction of RelA(-/-) B cells with either cFLIP or Bcl-2 significantly reduced TNF-alpha killing. Together, these results indicate that RelA plays a crucial role in regulating developing B cell survival by inhibiting TNF-alpha cytotoxicity. PMID- 14530315 TI - Highly biased type 1 immune responses in mice deficient in LFA-1 in Listeria monocytogenes infection are caused by elevated IL-12 production by granulocytes. AB - LFA-1 (CD11a/CD18) plays a key role in various inflammatory responses. Here we show that the acquired immune response to Listeria monocytogenes is highly biased toward type 1 in the absence of LFA-1. At the early stage of listeriosis, numbers of IFN-gamma producers in the liver and spleen of LFA-1(-/-) mice were markedly increased compared with heterozygous littermates and Valpha14(+)NKT cell deficient mice, and NK cells were major IFN-gamma producers. Numbers of IL-12 producers were also markedly elevated in LFA-1(-/-) mice compared with heterozygous littermates, and endogenous IL-12 neutralization impaired IFN-gamma production by NK cells. Granulocyte depletion diminished numbers of IL-12 producers and IFN-gamma-secreting NK cells in the liver of LFA-1(-/-) mice. Granulocytes from the liver of L. monocytogenes-infected LFA-1(-/-) mice were potent IL-12 producers. Thus, in the absence of LFA-1, granulocytes are a major source of IL-12 at the early stage of listeriosis. We assume that highly biased type 1 immune responses in LFA-1(-/-) mice are caused by increased levels of IL 12 from granulocytes and that granulocytes play a major role in IFN-gamma secretion by NK cells. In conclusion, LFA-1 regulates type 1 immune responses by controlling prompt infiltration of IL-12-producing granulocytes into sites of inflammation. PMID- 14530317 TI - Burn injury promotes antigen-driven Th2-type responses in vivo. AB - Severe injury induces detrimental changes in immune function, often leaving the host highly susceptible to developing life-threatening opportunistic infections. Advances in our understanding of how injury influences host immune responses suggest that injury causes a phenotypic imbalance in the regulation of Th1- and Th2-type immune responses. We report in this study, using a TCR transgenic CD4(+) T cell adoptive transfer approach, that injury skews T cell responses toward increased Th2-type reactivity in vivo without substantially limiting Ag-driven CD4(+) T cell expansion. The increased Th2-type response did not occur unless injured mice were immunized with specific Ag, suggesting that the phenotypic switch is Ag dependent. These findings establish that severe injury induces fundamental changes in the induction of Ag-specific CD4(+) Th cell responses favoring the development of Th2-type immune reactivity in vivo. PMID- 14530316 TI - Expression and function of Toll-like receptors in eosinophils: activation by Toll like receptor 7 ligand. AB - We investigated the expression of a panel of Toll-like receptors (TLRs) and their functions in human eosinophils. Eosinophils constitutively expressed TLR1, TLR4, TLR7, TLR9, and TLR10 mRNAs (TLR4 greater than TLR1, TLR7, TLR9, and TLR10 greater than TLR6). In contrast, neutrophils expressed a larger variety of TLR mRNAs (TLR1, TLR2, TLR4, TLR6, TLR8 greater than TLR5, TLR9, and TLR10 greater than TLR7). Although the expression levels in eosinophils were generally less prominent compared with those in neutrophils, eosinophils expressed a higher level of TLR7. Furthermore, among various TLR ligands (S-(2,3-bis(palmitoyloxy) (2-RS)-propyl)-N-palmitoyl-Cys-Ser-(Lys)(4), poly(I:C), LPS, R-848, and CpG DNA), only R-848, a ligand of TLR7 and TLR8, regulated adhesion molecule (CD11b and L selectin) expression, prolonged survival, and induced superoxide generation in eosinophils. Stimulation of eosinophils by R-848 led to p38 mitogen-activated protein kinase activation, and SB203580, a p38 mitogen-activated protein kinase inhibitor, almost completely attenuated R-848-induced superoxide generation. Although TLR8 mRNA expression was hardly detectable in freshly isolated eosinophils, mRNA expression of TLR8 as well as TLR7 was exclusively up-regulated by IFN-gamma but not by either IL-4 or IL-5. The up-regulation of the TLRs by IFN gamma had potentially functional significance: the extent of R-848-induced modulation of adhesion molecule expression was significantly greater in cells treated with IFN-gamma compared with untreated cells. Although the natural ligands for TLR7 and TLR8 have not yet been identified, our results suggest that eosinophil TLR7/8 systems represent a potentially important mechanism of a host defensive role against viral infection and mechanism linking exacerbation of allergic inflammation and viral infection. PMID- 14530319 TI - Broadly increased sensitivity to cytotoxic T lymphocytes resulting from Nef epitope escape mutations. AB - Nef is an HIV-1 protein that is absent in most retroviruses, yet its reading frame is highly maintained despite frequent targeting by CD8(+) CTL in vivo. Because Nef is not necessarily required for viral replication, this consistent maintenance suggests that Nef plays an important role(s) and substantial fitness constraints prevent its loss in vivo. The ability of Nef to down-regulate cell surface MHC class I (MHC-I) molecules and render infected cells resistant to CTL in general is likely to be an important contributing function. We demonstrate that mutational escape of HIV-1 from Nef-specific CTL in vitro leads to progeny virions that are increased in their susceptibility to CTL of specificities for proteins other than Nef. The escape mutants contain multiple nef mutations that impair the ability of the virus to down-regulate MHC-I through disruption of its reading frame as well as epitope point mutations. Given the rarity of nef frameshifts in vivo, these data support the concept that the ability to down regulate MHC-I could be a key constraint for preservation of Nef in vivo. PMID- 14530318 TI - Th1 cytokines regulate adenosine receptors and their downstream signaling elements in human microvascular endothelial cells. AB - We and others have shown that adenosine, acting at its receptors, is a potent modulator of inflammation and angiogenesis. To better understand the regulation of adenosine receptors during these processes we studied the effects of IL-1, TNF alpha, and IFN-gamma on expression and function of adenosine receptors and select members of their coupling G proteins in human dermal microvascular endothelial cells (HMVEC). HMVEC expressed message and protein for A(2A) and A(2B), but not A(1) or A(3) receptors. IL-1 and TNF-alpha treatment increased message and protein expression of A(2A) and A(2B) receptor. IFN-gamma treatment also increased the expression of A(2B) receptors, but decreased expression of A(2A) receptors. Resting HMVEC and IFN-gamma-treated cells showed minimal cAMP response to the selective A(2A) receptor agonist 2-[2-(4-chlorophenyl)ethoxy]adenosine (MRE0094). In contrast, MRE0094 stimulated a dose-dependent increase in cAMP levels in TNF-alpha-treated cells that was almost completely blocked by the A(2A) receptor antagonist ZM-241385 (4-[2-[7-amino-2-(2-furyl)[1,2,4]triazolo-[2,3 a][1,3,5]triazin-5-ylamino]ethyl]phenol). The nonselective adenosine receptor agonist 5'-(N-ethylcarboxamido)adenosine increased cAMP levels in both TNF-alpha- and IFN-gamma-treated cells, but not control cells, and its effect was only partially reversed by ZM-241385 in TNF-alpha-treated cells and not affected in IFN-gamma-treated cells. HMVEC expressed a higher level of G protein beta1 isoform than beta4 isoform. Although none of the cytokines tested affected G(beta1) expression, both IL-1 and TNF-alpha significantly up-regulated G(beta4) expression. These findings indicate that inflammatory cytokines modulate adenosine receptor expression and function on HMVECs and suggest that the interaction between proinflammatory cytokines and adenosine receptors may affect therapeutic responses to anti-inflammatory drugs that act via adenosine-dependent mechanisms. PMID- 14530320 TI - MHC class I gene conversion mutations alter the CD8 T cell repertoire. AB - MHC class I molecules are highly polymorphic within populations. This diversity is thought to be the result of selective maintenance of new class I alleles formed by gene conversion. It has been proposed that rare alleles are maintained by their ability to confer resistance to common pathogens. Investigation has focused on differences in the presentation of foreign Ags by class I alleles, but the majority of peptides presented by class I molecules are self peptides used in shaping the naive T cell repertoire. We propose that the key substrate for the natural selection of class I gene conversion variants is the diversity in immune potential formed by new alleles. We show that T cells compete with each other for niches in the thymus and spleen during development, and that competition between different clones is dramatically affected by class I mutations. We also show that peripheral naive T cells proliferate preferentially in the presence of the class I variant that directed T cell development. The data argue that class I gene conversion mutations dramatically affect both the development and the maintenance of the naive CD8 T cell repertoire. PMID- 14530321 TI - Coordinate regulation of lymphocyte-endothelial interactions by pregnancy associated hormones. AB - Precursors of uterine NK cells home to the uterus during early pregnancy from multiple lymphohemopoietic sources. In mouse uterine tissue, pregnancy markedly up-regulates both L-selectin- and alpha(4) integrin-dependent adhesion pathways for circulating human CD56(bright) cells, the phenotype of human uterine NK cells. Based on roles for these adhesion molecules in lymphocyte homing, we examined effects of pregnancy or the steroid hormones 17beta-estradiol or progesterone on lymphocyte-endothelial interactions in secondary lymphoid tissues and in uterus. From preimplantation gestation day 3, specialized high endothelial venules in peripheral lymph nodes and Peyer's patches supported elevated L selectin and alpha(4)beta(7) integrin-dependent lymphocyte adhesion under shear throughout pregnancy, as compared with high endothelial venules of virgin or postpartum donors. Squamous endothelium from nonlymphoid tissue was not affected. Pregnancy-equivalent endothelial responses were observed in lymph nodes and Peyer's patches from ovariectomized mice receiving 17beta-estradiol and/or progesterone replacement therapy. Adhesion of human CD56(bright) cells to uteri from pregnant or hormone-treated ovariectomized mice was enhanced through L selectin- and alpha(4) integrin-dependent mechanisms and involved multiple vascular adhesion molecules including mucosal addressin cell adhesion molecule-1, VCAM-1, and peripheral lymph node addressin. Analysis of Tie2-green fluorescence protein transgenic mice demonstrated that CD56(bright) cells adhered primarily to vascular endothelium within the decidua basalis. Microdomain localization of adhesion involving large clusters of lymphocytes was induced on uteri from natural matings, but not pseudopregnancy. Steroid hormones also had independent effects on L-selectin function in splenic lymphocytes that mimicked physiological stimulation induced by pregnancy or fever-range temperatures. These results provide the first evidence for coordinated, organ-specific, steroid hormone induced changes in lymphocyte homing mechanisms that could contribute to local and systemic immune responses during pregnancy. PMID- 14530322 TI - Glycolipid antigen drives rapid expansion and sustained cytokine production by NK T cells. AB - NKT cells are enigmatic lymphocytes that respond to glycolipid Ags presented by CD1d. Although they are key immunoregulatory cells, with a critical role in immunity to cancer, infection, and autoimmune diseases, little is known about how they respond to antigenic challenge. Current theories suggest that NKT cells die within hours of stimulation, implying that their direct impact on the immune system derives from the initial cytokine burst released before their death. Here we show that NKT cell disappearance results from TCR down-regulation rather than apoptosis, and that they expand to many times their normal number in peripheral tissues within 2-3 days of stimulation, before contracting to normal numbers over subsequent days. This expansion is associated with ongoing cytokine production, biased toward a Th1 (IFN-gamma(+) IL-4(-)) phenotype, in contrast to their initial Th0 (IFN-gamma(+)IL-4(+)) phenotype. This study provides critical new insight into how NKT cells can have such a major impact on immune responses, lasting many days beyond the initial stimulation of these cells. PMID- 14530323 TI - Immunization with Th-CTL fusion peptide and cytosine-phosphate-guanine DNA in transgenic HLA-A2 mice induces recognition of HIV-infected T cells and clears vaccinia virus challenge. AB - We evaluated immunogenicity of a novel Th-CTL fusion peptide composed of the pan DR Th epitope and a CTL epitope derived from HIV-pol in two transgenic HLA A*0201/K(b) mouse models. The immunogenicity of peptides of this structure is highly dependent on coadministered cytosine-phosphate-guanine DNA. Initial evaluations of peptide-specific immunity are based on results of chromium release assay, intracellular cytokine, and tetramer staining. Significant cytotoxic T cell responses are found upon a single immunization with as low as 0.1 nmol both peptide and cytosine-phosphate-guanine DNA. Splenocytes from immunized mice recognize naturally processed HIV-pol expressed from vaccinia virus (pol-VV). Translation of immunologic criteria into more relevant assays was pursued using systemic challenge of immunized mice with pol-VV. Only mice receiving both peptide and DNA together successfully cleared upward of 6 logs of virus from ovaries, compared with controls. Challenge with pol-VV by intranasal route of intranasal immunized mice showed a significant reduction in the levels of VV in lung compared with naive mice. A convincing demonstration of the relevance of these vaccines is the robust lysis of HIV-infected Jurkat T cells (JA2/R7/Hyg) by immune splenocytes from peptide- and DNA-immunized mice. This surprisingly effective immunization merits consideration for clinical evaluation, because it succeeded in causing immune recognition and lysis of cells infected with its target virus and reduction in titer of highly pathogenic VV. PMID- 14530324 TI - Dynamics of pathogenic and suppressor T cells in autoimmune diabetes development. AB - In the nonobese diabetic (NOD) mouse, pathogenic and suppressor CD4(+) T cells can be distinguished by the constitutive expression of CD25. In this study, we demonstrate that the progression of autoimmune diabetes in NOD mice reflects modifications in both T cell subsets. CD4(+)CD25(+) suppressor T cells from 8-, but not 16-wk-old NOD mice delayed the onset of diabetes transferred by 16-wk-old CD25-depleted spleen cells. These results were paralleled by the inhibition of alloantigen-induced proliferation of CD4(+)CD25(-) cells, indicating an age dependent decrease in suppressive activity. In addition, CD4(+)CD25(-) pathogenic T cells became progressively less sensitive to immunoregulation by CD4(+)CD25(+) T cells during diabetes development. CD4(+)CD25(-) T cells showed a higher proliferation and produced more IFN-gamma, but less IL-4 and IL-10, whereas CD4(+)CD25(+) T suppressor cells produced significantly lower levels of IL-10 in 16- compared with 8-wk-old NOD mice. Consistent with these findings, a higher frequency of Th1 cells was observed in the pancreas of 16-wk-old compared with 8 wk-old NOD mice. An increased percentage of CD4(+)CD25(-) T cells expressing CD54 was present in 16-wk-old and in diabetic NOD, but not in BALB/c mice. Costimulation via CD54 increased the proliferation of CD4(+)CD25(-) T cells from 16-, but not 8-wk-old NOD mice, and blocking CD54 prevented their proliferation, consistent with the role of CD54 in diabetes development. Thus, the pathogenesis of autoimmune diabetes in NOD mice is correlated with both an enhanced pathogenicity of CD4(+)CD25(-) T cells and a decreased suppressive activity of CD4(+)CD25(+) T cells. PMID- 14530325 TI - The central tolerance response to male antigen in normal mice is deletion and not receptor editing. AB - It is widely accepted that developing T cells can undergo clonal deletion in the thymus in response to a high affinity self-Ag. This is largely based on studies of TCR transgenics. However, encounter with high affinity self-Ag can also result in receptor editing in TCR transgenic models. Because all TCR transgenics display ectopic receptor expression, the tolerance mechanism that predominates in normal mice remains an open question. When self-Ag drives receptor editing during T cell development, one expects to find in-frame, self-reactive TCRalpha joins on TCR excision circles (TRECs), which are the products of secondary V/J recombination in the TCRalpha locus. Such joins are not expected if clonal deletion occurs, because the progenitor cell would be eliminated by apoptosis. To test the relative utilization of receptor editing vs clonal deletion, we determined the frequency of in-frame, male-specific joins on TRECs in male and female HYbeta transgenic mice. In comparison with female HYbeta transgenic mice, our analysis showed a lower frequency of TRECs with male-reactive V17J57 joins in male mice. Thus, it would appear that receptor editing is not a predominant tolerance mechanism for this self-Ag. PMID- 14530326 TI - HSP110-HER2/neu chaperone complex vaccine induces protective immunity against spontaneous mammary tumors in HER-2/neu transgenic mice. AB - Heat shock proteins (HSPs) are shown to be strong immunoadjuvants, eliciting both innate and adaptive immune responses against cancers. HSP110 is related in sequence to HSP70 and is approximately 4-fold more efficient in binding to and stabilizing denatured protein substrates compared with HSP70. In the present study we evaluated the ability of a heat shock complex of HSP110 with the intracellular domain (ICD) of human HER-2/neu to elicit effective antitumor immune responses and to inhibit spontaneous mammary tumors in FVB-neu (FVBN202) transgenic mice. The HSP110-ICD complex was capable of breaking tolerance against the rat neu protein and inhibiting spontaneous mammary tumor development. This vaccine induced ICD-specific IFN-gamma and IL-4 production. Depletion studies revealed that CD8(+) T cells were involved in protection against challenge with mouse mammary tumors, whereas CD4(+) T cells revealed partial protection. Increased IgG2a Ab titer in the sera of tumor-free animals after vaccination and elevated CD4(+) CD25(+) regulatory T cells in the PBL of tumor-bearing animals suggested that IFN-gamma-producing Th1 cells may be responsible for partial protection of CD4(+) T cells against the mammary tumor challenge, whereas CD4(+)CD25(+) regulatory T cells (Th2 cells) may suppress the antitumor immune responses. Together, these results suggest that HSP110-ICD complex can elicit effective IFN-gamma-producing T cells against spontaneous mammary tumors and that up-regulation of CD4(+) CD25(+) regulatory T cells may prevent complete eradication of the tumor following immunotherapy. PMID- 14530327 TI - The tetraspanin CD81 regulates the expression of CD19 during B cell development in a postendoplasmic reticulum compartment. AB - CD81 is a widely expressed tetraspanin that associates in B cells with CD19 in the CD19-CD21-CD81 signaling complex. CD81 is necessary for normal CD19 expression; cd81(-/-) B cells express lower levels of CD19, especially cd81(-/-) small pre-BII cells, which are almost devoid of surface CD19. The dependence of CD19 expression on CD81 is specific to this particular tetraspanin since cd9(-/-) B cells express normal levels of CD19. Furthermore, expression of human CD81 in mouse cd81(-/-) B cells restored surface CD19 to normal levels. Quantitative analysis of CD19 mRNA demonstrated normal levels, even in cd81(-/-) pre-BII cells. Analysis of CD19 at the protein level identified two CD19 glycoforms in both wild-type and cd81(-/-) B cells. The higher M(r) glycoform is significantly reduced in cd81(-/-) B cells and is endoglycosidase H (endo-H) resistant. In contrast, the low M(r) glycoform is comparably expressed in cd81(-/-) and in wild type B cells and is endo-H sensitive. Because endo-H sensitivity is tightly correlated with endoplasmic reticulum localization, we suggest that the dependency of CD19 expression on CD81 occurs in a postendoplasmic reticulum compartment where CD81 is necessary for normal trafficking or for surface membrane stability of CD19. PMID- 14530328 TI - Psychological stress exerts an adjuvant effect on skin dendritic cell functions in vivo. AB - Psychological stress affects the pathophysiology of infectious, inflammatory, and autoimmune diseases. However, the mechanisms by which stress could modulate immune responses in vivo are poorly understood. In this study, we report that application of a psychological stress before immunization exerts an adjuvant effect on dendritic cell (DC), resulting in increased primary and memory Ag specific T cell immune responses. Acute stress dramatically enhanced the skin delayed-type hypersensitivity reaction to haptens, which is mediated by CD8(+) CTLs. This effect was due to increased migration of skin DCs, resulting in augmented CD8(+) T cell priming in draining lymph nodes and enhanced recruitment of CD8(+) T cell effectors in the skin upon challenge. This adjuvant effect of stress was mediated by norepinephrine (NE), but not corticosteroids, as demonstrated by normalization of the skin delayed-type hypersensitivity reaction and DC migratory properties following selective depletion of NE. These results suggest that release of NE by sympathetic nerve termini during a psychological stress exerts an adjuvant effect on DC by promoting enhanced migration to lymph nodes, resulting in increased Ag-specific T cell responses. Our findings may open new ways in the treatment of inflammatory diseases, e.g., psoriasis, allergic contact dermatitis, and atopic dermatitis. PMID- 14530329 TI - T cells induce extended class II MHC compartments in dendritic cells in a Toll like receptor-dependent manner. AB - Interaction of Ag-loaded dendritic cells with Ag-specific CD4 T cells induces the formation of long tubular class II MHC-positive compartments that polarize toward the T cell. We show involvement of a Toll-like receptor-mediated signal in this unusual form of intracellular class II MHC trafficking. First, wild-type dendritic cells loaded with LPS-free Ag failed to show formation of class II positive tubules upon Ag-specific T cell engagement, but did so upon supplementation of the Ag with low concentrations of LPS. Second, Ag-loaded myeloid differentiation factor 88 -deficient dendritic cells failed to form these tubules upon interaction with T cells, regardless of the presence of LPS. Finally, inclusion of a cell-permeable peptide that blocks TNFR-associated factor 6 function, downstream of myeloid differentiation factor 88, blocked T cell dependent tubulation. A Toll-like receptor-dependent signal is thus required to allow Ag-loaded dendritic cells to respond to T cell contact by formation of extended endosomal compartments. This activation does not result in massive translocation of class II MHC molecules to the cell surface. PMID- 14530330 TI - Functional segregation of the TCR and antigen-MHC complexes on the surface of CTL. AB - As CTL adhere to and lyze their targets, they extract cognate Ag-MHC and represent this on their own cell surface. Whether such self-presented cognate Ag stimulate the TCR of a CTL is uncertain. To analyze this, we examined TCR capping in response to self-presented Ag. We found that OVA peptide-specific OT-1 CTL that were pulsed with cognate peptide Ag did not cap their TCR, implying that the autologously presented MHC-Ag complex does not normally stimulate the TCR. However, this functional separation of the TCR and its ligand on the cell surface was not absolute. Treatment of Ag-pulsed OT-1 CTL with agents that alter cell surface charge, including trypsin, papain, tunicamycin, neuraminidase, and polybrene, allowed Ag-specific TCR capping. The TCR capped together with the restricting MHC molecule on the surface of the cell, implying an interaction between the TCR and cell-associated Ag. Further, the treated CTL underwent a time and dose-dependent suicidal death that was both Fas- and perforin-dependent. Therefore, our results indicate that the association of the TCR with its MHC peptide ligand on the surface of a CTL is normally proscribed by biophysical properties of the plasma membrane. Overcoming this restriction allows TCR stimulation and induces CTL effector functions and cell suicide. PMID- 14530331 TI - T cell development in mice expressing CD1d directed by a classical MHC class II promoter. AB - CD1d and nonclassical MHC molecules differ markedly from classical MHC ligands in their ability to promote the selection and differentiation of developing T cells. Whereas classical MHC-restricted T cells have a predominantly naive phenotype and a broad TCR repertoire, most other T cells have a memory and/or NKT phenotype with a restricted repertoire. Because the nonclassical ligands selecting these memory-type cells are expressed by bone marrow-derived cells, it has been suggested that the development of large repertoires of naive-type cells was dependent on the classical MHC expression pattern in the thymus cortex, high on epithelial cells and low on cortical thymocytes. We redirected CD1d expression using the classical MHC II Ealpha promoter. pEalpha-CD1d mice lacked memory-type NKT cells, but, surprisingly, they did not acquire the reciprocal ability to select a diverse population of naive CD1d-restricted cells. These findings suggest that, whereas the development of NKT cells is dependent on the pattern of CD1d expression, the absence of a broad, naive CD1d-restricted T cell repertoire may reflect intrinsic limitations of the pool of TCR genes or lipid Ags. PMID- 14530332 TI - Expression of CD1d under the control of a MHC class Ia promoter skews the development of NKT cells, but not CD8+ T cells. AB - Although CD1d and MHC class Ia share similar overall structure, they have distinct levels and patterns of surface expression. While the expression of CD1d1 is known to be essential for the development of NKT cells, the contribution of CD1d1 to the development of CD8(+) T cells appears to be inconsequential. To investigate whether CD1d tissue distribution and expression levels confer differential capacity in selecting these two T cell subsets, we analyzed CD8 and NKT cell compartments in K(b)-CD1d-transgenic mice that lack endogenous MHC class Ia and CD1d, respectively. We found that MHC class Ia-like expression pattern and tissue distribution are not sufficient for CD1d to rescue the development of CD8(+) T cells, suggesting that unique structural features of CD1d preclude its active participation in selection of CD8(+) T cells. Conversely, cell type specific CD1d surface density is important for the selection of NKT cells, as the NKT cell compartment was only partially rescued by the K(b)-CD1d transgene. We have previously demonstrated that increased CD1d expression on dendritic cells enhanced negative selection of NKT cells. In this study, we show that cell type specific expression levels of CD1d establish a narrow window between positive and negative selection, suggesting that the distinct CD1d expression pattern may be selected evolutionarily to ensure optimal output of NKT cells. PMID- 14530333 TI - T cell-specific disruption of arylhydrocarbon receptor nuclear translocator (Arnt) gene causes resistance to 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced thymic involution. AB - The arylhydrocarbon receptor nuclear translocator (ARNT) is a member of the basic helix-loop-helix, PER-ARNT-SIM family of heterodimeric transcription factors, and serves as a dimerization partner for arylhydrocarbon receptor (AHR) and hypoxia inducible factor-1alpha. To assess the function of ARNT in T cells, we disrupted the Arnt gene specifically in T cells of mice by conditional gene targeting using T cell-specific p56(lck)-Cre (Lck-Cre) transgenic Arnt-floxed mice. Thus generated, T cell-specific Arnt-disrupted mice (Lck-Cre;Arnt(flox/Delta) transgenic mice) exhibited complete loss of the expression of ARNT protein only in T cells, and were viable and appeared normal. The Arnt-disrupted T cells in the thymus were phenotypically and histologically normal. The Arnt-deficient T cells in the spleen were capable of responding to TCR stimulation in vitro. However, unlike normal mice in which exposure to the environmental pollutant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), an AHR ligand, resulted in thymic involution, the thymus of Lck-Cre;Arnt(flox/Delta) mice were resistant to TCDD treatment in vivo. In contrast, benzo(a)pyrene, another AHR ligand, still caused thymic involution in Lck-Cre;Arnt(flox/Delta) mice. Finally, fetal thymus organ culture using Lck-Cre;Arnt(flox/Delta) and K5-Cre;Arnt(flox/Delta) (epithelial cell-specific Arnt-disrupted mice) showed that thymocytes rather than thymic epithelial cells are predominantly responsible for TCDD-induced thymic atrophy. Our results indicate that ARNT in T lineage cells is essential for TCDD-mediated thymic involution. PMID- 14530334 TI - Enhanced effector and memory CTL responses generated by incorporation of receptor activator of NF-kappa B (RANK)/RANK ligand costimulatory molecules into dendritic cell immunogens expressing a human tumor-specific antigen. AB - The outcome of dendritic cell (DC) presentation of Ag to T cells via the TCR/MHC synapse is determined by second signaling through CD80/86 and, importantly, by ligation of costimulatory ligands and receptors located at the DC and T cell surfaces. Downstream signaling triggered by costimulatory molecule ligation results in reciprocal DC and T cell activation and survival, which predisposes to enhanced T cell-mediated immune responses. In this study, we used adenoviral vectors to express a model tumor Ag (the E7 oncoprotein of human papillomavirus 16) with or without coexpression of receptor activator of NF-kappaB (RANK)/RANK ligand (RANKL) or CD40/CD40L costimulatory molecules, and used these transgenic DCs to immunize mice for the generation of E7-directed CD8(+) T cell responses. We show that coexpression of RANK/RANKL, but not CD40/CD40L, in E7-expressing DCs augmented E7-specific IFN-gamma-secreting effector and memory T cells and E7 specific CTLs. These responses were also augmented by coexpression of T cell costimulatory molecules (RANKL and CD40L) or DC costimulatory molecules (RANK and CD40) in the E7-expressing DC immunogens. Augmentation of CTL responses correlated with up-regulation of CD80 and CD86 expression in DCs transduced with costimulatory molecules, suggesting a mechanism for enhanced T cell activation/survival. These results have generic implications for improved tumor Ag-expressing DC vaccines, and specific implications for a DC-based vaccine approach for human papillomavirus 16-associated cervical carcinoma. PMID- 14530335 TI - Clonal expansion of double-positive intraepithelial lymphocytes by MHC class I related chain A expressed in mouse small intestinal epithelium. AB - Expression of a distant homologue MHC class I molecule, MHC class I-related chain A (MICA), has been found to be stress inducible and limited to the intestinal epithelium. This nonclassical MHC molecule is associated with various carcinomas in humans. To understand the biological consequences of MICA expression in the gut, we generated transgenic (Tg) mice (T3(b)-MICA Tg) under the control of the T3(b) promoter. The T3(b)-MICA Tg mice expressed MICA selectively in the intestine and had an increased number of TCRalphabeta CD4CD8alphaalpha, double positive (DP) intraepithelial lymphocytes (IELs) in the small bowel. These MICA expanded DP IELs exhibited a bias to Vbeta8.2 and overlapped motifs of the complementarity-determining region 3 region among various Tg mice. Hence, the overexpression of MICA resulted in a clonal expansion of DP IELs. Studies in model of inflammatory bowel disease showed that transgenic MICA was able to attenuate the acute colitis induced by dextran sodium sulfate administration. Therefore, this unique in vivo model will enable investigation of possible influences of stress-inducible MICA on the gut immune surveillance. PMID- 14530336 TI - Clustering of Th cell epitopes on exposed regions of HIV envelope despite defects in antibody activity. AB - A long-standing question in the field of immunology concerns the factors that contribute to Th cell epitope immunodominance. For a number of viral membrane proteins, Th cell epitopes are localized to exposed protein surfaces, often overlapping with Ab binding sites. It has therefore been proposed that Abs on B cell surfaces selectively bind and protect exposed protein fragments during Ag processing, and that this interaction helps to shape the Th cell repertoire. While attractive in concept, this hypothesis has not been thoroughly tested. To test this hypothesis, we have compared Th cell peptide immunodominance in normal C57BL/6 mice with that in C57BL/6( micro MT/ micro MT) mice (lacking normal B cell activity). Animals were first vaccinated with DNA constructs expressing one of three different HIV envelope proteins, after which the CD4(+) T cell response profiles were characterized toward overlapping peptides using an IFN-gamma ELISPOT assay. We found a striking similarity between the peptide response profiles in the two mouse strains. Profiles also matched those of previous experiments in which different envelope vaccination regimens were used. Our results clearly demonstrate that normal Ab activity is not required for the establishment or maintenance of Th peptide immunodominance in the HIV envelope response. To explain the clustering of Th cell epitopes, we propose that localization of peptide on exposed envelope surfaces facilitates proteolytic activity and preferential peptide shuttling through the Ag processing pathway. PMID- 14530337 TI - Lysosomal localization of murine CD1d mediated by AP-3 is necessary for NK T cell development. AB - The presentation of lipid and glycolipid Ags to T cells is mediated through CD1 molecules. In the mouse and rat only a single isoform, CD1d, performs these functions, while humans and all other mammals studied have members of both group I (CD1a, -b, and -c) and group II (CD1d) isoforms. Murine CD1d contains a cytoplasmic tyrosine-based sorting motif that is similar to motifs recognized by adaptor protein complexes that sort transmembrane proteins. Here we show that the adaptor protein complex, AP-3, directly interacts with murine CD1d and controls its targeting to lysosomes. AP-3 deficiency results in a redistribution of CD1d from lysosomes to the cell surface of thymocytes, B cell-depleted splenocytes, and dendritic cells. The altered trafficking of CD1d in AP-3-deficient mice results in a significant reduction of NK1.1(+)TCR-beta(+) and CD1d tetramer positive cells, consistent with a defect in CD1d self-Ag presentation and thymocyte-positive selection. The AP-3 complex has recently been shown to associate with the human CD1b isoform, which has an intracellular distribution pattern similar to that of murine CD1d. We propose that lysosomal sampling may be so critical for efficient host defense that mice have evolved mechanisms to target their single CD1 isoform to lysosomes for sampling lipid Ags. Here we show the dominant mechanism for this trafficking is mediated by AP-3. PMID- 14530338 TI - Blockade of B7-H1 suppresses the development of chronic intestinal inflammation. AB - A newly identified costimulatory molecule, programmed death-1 (PD-1), provides a negative signal that is essential for immune homeostasis. However, it has been suggested that its ligands, B7-H1 (PD-L1) and B7-dendritic cells (B7-DC; PD-L2), could also costimulate T cell proliferation and cytokine secretion. Here we demonstrate the involvement of PD-1/B7-H1 and B7-DC interaction in the development of colitis. We first examined the expression profiles of PD-1 and its ligands in both human inflammatory bowel disease and a murine chronic colitis model induced by adoptive transfer of CD4(+)CD45RB(high) T cells to SCID mice. Second, we assessed the therapeutic potential of neutralizing anti-B7-H1 and/or B7-DC mAbs using this colitis model. We found significantly increased expression of PD-1 on T cells and of B7-H1 on T, B, and macrophage/DCs in inflamed colon from both inflammatory bowel disease patients and colitic mice. Unexpectedly, the administration of anti-B7-H1, but not anti-B7-DC, mAb after transfer of CD4(+)CD45RB(high) T cells suppressed wasting disease with colitis, abrogated leukocyte infiltration, and reduced the production of IFN-gamma, IL-2, and TNF alpha, but not IL-4 or IL-10, by lamina propria CD4(+) T cells. These data suggest that the interaction of PD-1/B7-H1, but not PD-1/B7-DC, might be involved in intestinal mucosal inflammation and also show a possible role of interaction between B7-H1 and an as yet unidentified receptor for B7-H1 in inducing T cell activation. PMID- 14530339 TI - Exposure of human primary colon carcinoma cells to anti-Fas interactions influences the emergence of pre-existing Fas-resistant metastatic subpopulations. AB - Fas, an important death receptor-mediated signaling pathway, has been shown to be down-regulated during human colon tumorigenesis; however, how alterations in Fas expression influence the metastatic process remains unresolved. In mouse models, loss of Fas function was found to be both necessary and sufficient for tumor progression. In this study, we investigated the link between functional Fas status and malignant phenotype using a matched pair of naturally occurring primary (Fas-sensitive) and metastatic (Fas-resistant) human colon carcinoma cell lines in both in vitro and in vivo (xenograft) settings. Metastatic sublines were produced in vitro from the primary tumor cell line by functional elimination of Fas-responsive cells. Conversely, sublines derived from the primary tumor in vivo at distal metastatic sites were Fas-resistant. In contrast, simply disrupting the Fas pathway by molecular-based strategies in the Fas-sensitive primary tumor failed to achieve the same metastatic outcome. Interestingly, both in vitro- and in vivo-produced sublines resembled the naturally occurring metastatic population, based on functional and morphologic studies and genome-scale gene expression profiling. Overall, using this human colon carcinoma model, we: 1) showed that loss of Fas function was linked to, but alone was insufficient for, acquisition of a detectable metastatic phenotype; 2) demonstrated that metastatic subpopulations pre-existed within the heterogeneous primary tumor, and that anti Fas interactions served as a selective pressure for their outgrowth; and 3) identified a large set of differentially expressed genes distinguishing the primary from metastatic malignant phenotypes. Thus, Fas-based interactions may represent a novel mechanism for the biologic or immunologic selection of certain types of Fas-resistant neoplastic clones with enhanced metastatic ability. PMID- 14530340 TI - Ex vivo analysis of thymic CD4 T cells in nonobese diabetic mice with tetramers generated from I-A(g7)/class II-associated invariant chain peptide precursors. AB - The MHC determines susceptibility and resistance to type 1 diabetes in humans and nonobese diabetic (NOD) mice. To investigate how a disease-associated MHC molecule shapes the T cell repertoire in NOD mice, we generated a series of tetramers from I-A(g7)/class II-associated invariant chain peptide precursors by peptide exchange. No CD4 T cell populations could be identified for two glutamic acid decarboxylase 65 peptides, but tetramers with a peptide mimetic recognized by the BDC-2.5 and other islet-specific T cell clones labeled a distinct population in the thymus of young NOD mice. Tetramer-positive cells were identified in the immature CD4(+)CD8(low) population that arises during positive selection, and in larger numbers in the more mature CD4(+)CD8(-) population. Tetramer labeling was specific based on the use of multiple control tetramers, including one with a single amino acid analog peptide in which a critical TCR contact residue was substituted. The T cell population was already present in the thymus of 2-wk-old NOD mice before the typical onset of insulitis and was detected in B10 mice congenic for the NOD MHC locus, but not B10 control mice. These results demonstrate that a T cell population can expand in the thymus of NOD mice to levels that are at least two to three orders of magnitude higher than estimated for a given specificity in the naive T cell pool. Based on these data, we propose a model in which I-A(g7) confers susceptibility to type 1 diabetes by biasing positive selection in the thymus and later presenting peptides from islet autoantigens to such T cells in the periphery. PMID- 14530341 TI - IFN-gamma-induced MHC class II expression: transactivation of class II transactivator promoter IV by IFN regulatory factor-1 is regulated by protein kinase C-alpha. AB - Previous studies based on pharmacological evidence suggested a requirement for protein kinase C (PKC) activity in the regulation of IFN-gamma-induced MHC class II (MHC-II) expression. In the present study, we investigated the molecular mechanisms by which PKC-alpha modulates IFN-gamma-induced MHC-II expression in the mouse macrophage cell line RAW 264.7. Overexpression of a dominant-negative (DN) mutant of PKC-alpha inhibited the expression of IFN-gamma-induced MHC-II but had no effect on IFN-gamma-induced STAT1 nuclear translocation and DNA binding activity, as well as on the expression of inducible NO synthase, IFN consensus sequence binding protein, MHC class I, IFN regulatory factor (IRF)-1, and IFN gamma-inducible protein-10. Further analysis showed that IFN-gamma-induced expression of the MHC class II transactivator (CIITA), a transcriptional coactivator essential for MHC-II expression, was inhibited in DN PKC-alpha overexpressing cells. Studies with reporter constructs containing the promoter IV region of CIITA revealed that overexpression of a constitutively active mutant of PKC-alpha enhanced IRF-1, but not IRF-2, transcriptional activity. Furthermore, characterization of IRF-1 from both normal and DN PKC-alpha-overexpressing cells revealed differences in IRF-1 posttranslational modifications. Collectively, our data suggest a novel regulatory mechanism for IFN-gamma-induced MHC-II expression, whereby PKC regulates CIITA expression by selectively modulating the transcriptional activity of IRF-1. PMID- 14530342 TI - The Rac-activating toxin cytotoxic necrotizing factor 1 oversees NK cell-mediated activity by regulating the actin/microtubule interplay. AB - The cell cytoskeleton is widely acknowledged as a master for NK cell function. Specifically, actin filaments guide the NK cell binding to target cells, engendering the formation of the so-called immunological synapse, while microtubules direct the killer behavior. All these cytoskeleton-dependent activities are competently governed by the Rho GTPases, a family of regulatory molecules encompassing the three different subfamilies, Rho, Rac, and Cdc42. By using a Rac GTPase-activating bacterial protein toxin from Escherichia coli named cytotoxic necrotizing factor 1 (CNF1), we obtained results supporting the activation of Rac GTPase as a booster for effector cell-binding efficiency, recruitment ability, and, consequently, cytotoxicity. In particular, the augmented killer capacity of CNF1-treated NK cells was associated with the increased expression of certain cell adhesion or activation-associated molecules and the reshaping of the actin and microtubule networks. Importantly, CNF1 counteracted the activity exerted by toxins disrupting the cytoskeletal architecture. Hence, the activation of Rho GTPases, particularly Rac, induced by CNF1, appears to orchestrate a dynamic cross talk between microtubules and actin filaments, leading to a fruitful NK cell activity and polarization state. Our findings suggest that protein toxins might be viewed as modulators of NK cell cytotoxic activity and could possibly be regarded as useful pharmacological tools for certain Rho-linked immune diseases in the near future. PMID- 14530343 TI - Phosphatidylcholine-specific phospholipase C activity is necessary for the activation of STAT6. AB - It is well established that Janus kinase (JAK) tyrosine kinases play a key role in the activation of STAT6 by IL-4. In this study, we investigated additional molecules involved in this process. We previously found that IL-4 and TNF-alpha cooperate in the activation of STAT6 and NF-kappaB, suggesting that these transcription factors are regulated by common intracellular signaling pathways. To test this hypothesis, we analyzed the effect of known inhibitors of NF-kappaB on the activation of STAT6. We discovered that inhibitors of phosphatidylcholine specific phospholipase C (PC-PLC), but not other lipases, blocked the activation of STAT6 by IL-4. The activation of PC-PLC seems to be an early event in IL-4 signaling, because its inhibition abrogated JAK activation and STAT6 tyrosine phosphorylation. Interestingly, we found that the effects of pervanadate and sodium orthovanadate on STAT6 activation correspond to their effect on PC-PLC. Thus, pervanadate by itself activated PC-PLC, JAK, and STAT6, whereas sodium orthovanadate suppressed PC-PLC, JAK, and STAT6 activation by IL-4. We further found that PC-PLC activation is necessary but not sufficient to promote STAT6 activation, and therefore, additional intracellular pathways regulated by IL-4 and pervanadate may collaborate with PC-PLC to signal STAT6 activation. It has been reported that IL-4 signals PC-PLC activation; in this study, we provide evidence that this phospholipase plays a key role in IL-4 signaling. PMID- 14530344 TI - NF-kappa B regulates BCL3 transcription in T lymphocytes through an intronic enhancer. AB - Exposure to soluble protein Ags in vivo leads to abortive proliferation of responding T cells. In the absence of a danger signal, artificially provided by adjuvants, most responding cells die, and the remainder typically become anergic. The adjuvant-derived signals provided to T cells are poorly understood, but recent work has identified BCL3 as the gene, of those tested, with the greatest differential transcriptional response to adjuvant administration in vivo. As an initial step in analyzing transcriptional responses of BCL3 in T cells, we have identified candidate regulatory regions within the locus through their evolutionary conservation and by analysis of DNase hypersensitivity. An evolutionarily conserved DNase hypersensitive site (HS3) within intron 2 was found to act as a transcriptional enhancer in response to stimuli that mimic TCR activation, namely, PHA and PMA. In luciferase reporter gene constructs transiently transfected into the Jurkat T cell line, the HS3 enhancer can cooperate not only with the BCL3 promoter, but also with an exogenous promoter from herpes simplex thymidine kinase. Deletional analysis revealed that a minimal sequence of approximately 81 bp is required for full enhancer activity. At the 5' end of this minimal sequence is a kappaB site, as confirmed by EMSAs. Mutation of this site in the context of the full-length HS3 abolished enhancer activity. Cotransfection with NF-kappaB p65 expression constructs dramatically increased luciferase activity, even without stimulation. Conversely, cotransfection with the NF-kappaB inhibitor IkappaBalpha reduced activation. Together, these results demonstrate a critical role for NF-kappaB in BCL3 transcriptional up-regulation by TCR-mimetic signals. PMID- 14530345 TI - Transcriptional control of murine CD94 gene: differential usage of dual promoters by lymphoid cell types. AB - The CD94 gene product is involved in controlling NK cell activation, and is one of a family of immune receptors that is found in the NK gene complex in both humans and mice, adjacent to members of the NKG2 family. CD94 forms a heterodimeric complex with several members of the NKG2 family on the surface of NK, T, and NKT cells. These complexes recognize the nonclassical MHC class I molecules HLA-E and Qa-1(b) in humans and mice, respectively. The mechanism for cell type-specific expression of CD94 and other genes from the NK gene complex has not yet been elucidated. In the current study, we show that the murine CD94 gene has two promoters, one of which is upstream of a previously unidentified exon. We illustrate by quantitative real-time PCR that lymphoid cell types use these two promoters differentially and that the promoter usage seen in adult cells is already established during fetal development. We determined that the differential promoter usage by NK cells appears to be susceptible to perturbation, as both the murine NK cell line LNK, as well as cultured C57BL/6 NK cells showed altered promoter usage relative to fresh NK cells. Furthermore, the promoter activity observed in transfection assays did not correlate with expression of the endogenous CD94 gene, suggesting the involvement of chromatin structure/methylation in transcriptional regulation. Our detection of DNase I hypersensitive sites at the CD94 locus that are present only in a cell line expressing endogenous CD94 supports this hypothesis. PMID- 14530346 TI - Dual phosphorylation of phosphoinositide 3-kinase adaptor Grb2-associated binder 2 is responsible for superoxide formation synergistically stimulated by Fc gamma and formyl-methionyl-leucyl-phenylalanine receptors in differentiated THP-1 cells. AB - The class Ia phosphoinositide (PI) 3-kinase consisting of p110 catalytic and p85 regulatory subunits is activated by Tyr kinase-linked membrane receptors such as FcgammaRII through the association of p85 with the phosphorylated receptors or adaptors. The heterodimeric PI 3-kinase is also activated by G protein-coupled chemotactic fMLP receptors, and activation of the lipid kinase plays an important role in various immune responses, including superoxide formation in neutrophils. Although fMLP-induced superoxide formation is markedly enhanced in FcgammaRII primed neutrophils, the molecular mechanisms remain poorly characterized. In this study, we identified two Tyr-phosphorylated proteins, c-Cbl (Casitas B-lineage lymphoma) and Grb2-associated binder 2 (Gab2), as PI 3-kinase adaptors that are Tyr phosphorylated upon the stimulation of FcgammaRII in differentiated neutrophil-like THP-1 cells. Interestingly, Gab2 was, but c-Cbl was not, further Ser/Thr phosphorylated by fMLP. Thus, the adaptor Gab2 appeared to be dually phosphorylated at the Ser/Thr and Tyr residues through the two different types of membrane receptors. The Ser/Thr phosphorylation of Gab2 required the activation of extracellular signal-regulated kinase, and fMLP receptor stimulation indeed activated extracellular signal-regulated kinase in the cells. Enhanced superoxide formation in response to Fcgamma and fMLP was markedly attenuated when the Gab2 Ser/Thr phosphorylation was inhibited. These results show the importance of the dual phosphorylation of PI 3-kinase adaptor Gab2 for the enhanced superoxide formation in neutrophil-type cells. PMID- 14530347 TI - Receptor glycosylation regulates Ly-49 binding to MHC class I. AB - Murine NK cells express the Ly-49 family of class I MHC-binding receptors that control their ability to lyse tumor or virally infected host target cells. X-ray crystallography studies have identified two predominant contact sites (sites 1 and 2) that are involved in the binding of the inhibitory receptor, Ly-49A, to H 2D(d). Ly-49G2 (inhibitory) and Ly-49D (activating) are highly homologous to Ly 49A and also recognize H-2D(d). However, the binding of Ly-49D and G(2) to H 2D(d) is of lower affinity than Ly-49A. All Ly-49s contain N-glycosylation motifs; however, the importance of receptor glycosylation in Ly-49-class I interactions has not been determined. Ly-49D and G(2) contain a glycosylation motif (NTT (221-223)), absent in Ly-49A, adjacent to one of the proposed binding sites for H-2D(d) (site 2). The presence of a complex carbohydrate group at this critical site could interfere with class I binding. In this study, we are able to demonstrate for the first time that Ly-49D binds H-2D(d) in the presence of mouse beta(2)-microglobulin. We also demonstrate that glycosylation of the NTT (221-23) motif of Ly-49D inteferes with recognition of H-2D(d). Alteration of the Ly-49D NTT (221-23) motif to abolish glycosylation at this site resulted in enhanced H 2D(d) binding and receptor activation. Furthermore, glycosylation of Ly-49G2 at NTT (221-23) also reduces receptor binding to H-2D(d) tetramers. Therefore, the addition of complex carbohydrates to the Ly-49 family of receptors may represent a mechanism by which NK cells regulate affinity for host class I ligands. PMID- 14530348 TI - Hemozoin increases IFN-gamma-inducible macrophage nitric oxide generation through extracellular signal-regulated kinase- and NF-kappa B-dependent pathways. AB - NO overproduction has been suggested to contribute to the immunopathology related to malaria infection. Even though a role for some parasite molecules (e.g., GPI) in NO induction has been proposed, the direct contribution of hemozoin (HZ), another parasite metabolite, remains to be established. Therefore, we were interested to determine whether Plasmodium falciparum (Pf) HZ and synthetic HZ, beta-hematin, alone or in combination with IFN-gamma, were able to induce macrophage (Mphi) NO synthesis. We observed that neither Pf HZ nor synthetic HZ led to NO generation in B10R murine Mphi; however, they significantly increased IFN-gamma-mediated inducible NO synthase (iNOS) mRNA and protein expression, and NO production. Next, by investigating the transductional mechanisms involved in this cellular regulation, we established that HZ induces extracellular signal regulated kinase (ERK)1/2 mitogen-activated protein kinase phosphorylation as well as NF-kappaB binding to the iNOS promoter, and enhances the IFN-gamma dependent activation of both second messengers. Of interest, cell pretreatment with specific inhibitors against either NF-kappaB or the ERK1/2 pathway blocked the HZ + IFN-gamma-inducible NF-kappaB activity and significantly reduced the HZ dependent increase on IFN-gamma-mediated iNOS and NO induction. Even though selective inhibition of the Janus kinase 2/STAT1alpha pathway suppressed NO synthesis in response to HZ + IFN-gamma, HZ alone did not activate this signaling pathway and did not have an up-regulating effect on the IFN-gamma-induced Janus kinase 2/STAT1alpha phosphorylation and STAT1alpha binding to the iNOS promoter. In conclusion, our results suggest that HZ exerts a potent synergistic effect on the IFN-gamma-inducible NO generation in Mphi via ERK- and NF-kappaB-dependent pathways. PMID- 14530349 TI - Deficient anti-listerial immunity in the absence of perforin can be restored by increasing memory CD8+ T cell numbers. AB - Compared with wild-type (WT) mice, Listeria monocytogenes (LM)-vaccinated perforin-deficient (PKO) mice have elevated levels of CD8(+) T cell memory, but exhibit reduced levels of protection against virulent LM. In this study, Ag specific CD8(+) T cells from LM-vaccinated WT and PKO mice were used in adoptive transfer assays to determine the contribution of perforin-dependent cytolysis in protective immunity to LM. Perforin deficiency resulted in an approximately 5 fold reduction in the per-cell protective capacity of Ag-specific memory CD8(+) T cells that was not caused by differences in memory cell quality as measured by CD62L/CD27 expression, TCR repertoire use, functional avidity, differences in expansion of Ag-specific cells upon infection, or maintenance of memory levels over time. However, perforin-deficient CD8(+) T cells exhibited reduced in vivo cytotoxic function compared to WT CD8(+) T cells. Consistent with the existence of perforin-independent effector pathways, double-vaccinated PKO mice were as resistant to challenge with LM as single-vaccinated WT mice. Thus, increasing the number of memory CD8(+) T cells can overcome diminished per-cell protective immunity in the absence of perforin. PMID- 14530350 TI - Evidence for naturally acquired T cell-mediated mucosal immunity to Neisseria meningitidis. AB - Naturally acquired protective immunity against Neisseria meningitidis is thought to partially explain the disparity between the high levels of carriage in the human nasopharynx and the rare incidence of disease. To investigate this immunity to Neisseria meningitidis at the mucosal level, in vitro cellular responses to outer membrane vesicle preparations derived from this pathogen were examined using mononuclear cells from the palatine tonsils of adults and children. Characterization of these responses was achieved by depletion of CD45RA(+), CD45RO(+), and CD19(+) populations and outer membrane vesicles derived from isogenic mutants expressing different serosubtypes of the major outer membrane protein, porin A (PorA), no PorA and membrane preparations from a mutant with no LPS (LpxA(-)). The magnitude of cellular proliferative responses against the outer membrane vesicles were strongly associated with age and were largely T cell mediated, involving both CD45RO(+) and CD45RA(+) T cell phenotypes. Responses were not dependent on LPS but consisted of both PorA cross-specific and non-PorA dependent responses. Cellular immunity against Neisseria meningitidis was found to be frequently associated with systemic IgG Abs but was not associated with serum bactericidal Abs. For the first time our results demonstrate an age associated acquisition of mucosal T effector/memory cell responses to Neisseria meningitidis. This mucosal cellular immunity can be present in the absence of serum bactericidal Abs, a classical marker of protective immunity. PMID- 14530351 TI - Myosin autoimmunity is not essential for cardiac inflammation in acute Chagas' disease. AB - Infection with the protozoan parasite Trypanosoma cruzi leads to acute myocarditis that is accompanied by autoimmunity to cardiac myosin in susceptible strains of mice. It has been difficult to determine the contribution of autoimmunity to tissue inflammation, because other inflammatory mechanisms, such as parasite-mediated myocytolysis and parasite-specific immunity, are coincident during active infection. To begin to investigate the contribution of myosin autoimmunity to myocarditis, we selectively inhibited myosin autoimmunity by restoring myosin tolerance via injection of myosin-coupled splenocytes. This tolerization regimen suppressed the strong myosin-specific delayed-type hypersensitivity (DTH) that normally develops in infected mice, although it did not affect myosin-specific Ab production. Suppression of myosin autoimmunity had no effect on myocarditis or cardiac parasitosis. In contrast, myosin tolerization completely abrogated myocarditis in mice immunized with purified myosin, which normally causes severe autoimmune myocarditis. In this case, myosin-specific DTH and Ab production were significantly reduced. We also examined the contribution of T. cruzi-specific immunity to inflammation by injection of T. cruzi-coupled splenocytes before infection. This treatment reduced T. cruzi DTH, although there was no effect on parasite-specific Ab production. Interestingly, cardiac inflammation was decreased, cardiac parasitosis was significantly increased, and mortality occurred earlier in the parasite-tolerized animals. These results indicate that myosin-specific autoimmunity, while a potentially important inflammatory mechanism in acute and chronic T. cruzi infection, is not essential for inflammation in acute disease. They also confirm previous studies showing that parasite-specific cell-mediated immunity is important for myocarditis and survival of T. cruzi infection. PMID- 14530353 TI - Mucosally delivered E1-deleted adenoviral vaccine carriers induce transgene product-specific antibody responses in neonatal mice. AB - E1-deleted adenoviral vectors of the human serotype 5 (AdHu5) and the chimpanzee serotype 68 (AdC68) expressing the rabies virus glycoprotein (rab.gp) were tested for induction of transgene product-specific Abs upon intranasal or oral immunization of newborn mice. Both vectors induced Abs to rabies virus that could be detected in serum and from mucosal secretions. Serum rabies virus neutralizing Ab titers sufficed to protect neonatally vaccinated mice against a subsequent challenge with rabies virus. The efficacy of the AdHu5rab.gp vector given orally to newborn mice born to AdHu5 virus-immune dams was not impaired by maternally transferred Abs to the vaccine carrier. PMID- 14530352 TI - Functional characterization of class Ia- and non-class Ia-restricted Chlamydia reactive CD8+ T cell responses in humans. AB - CD8(+) T cells are a key immune component for the eradication of many intracellular pathogens. This study aims to characterize the human CD8(+) T cell response to naturally processed chlamydial Ags in individuals exposed to the intracellular pathogen Chlamydia trachomatis. By using C. trachomatis-infected autologous dendritic cells (DCs) as stimulators, Chlamydia-reactive CD8(+) T cell responses were detected in all 10 individuals tested. The majority of the Chlamydia-reactive CD8(+) T cells were non-MHC class Ia restricted in all three of the individuals tested. From one donor, three non-class Ia-restricted and two class Ia-restricted Chlamydia-specific CD8(+) T cells were cloned and characterized further. All five T cell clones secreted IFN-gamma in response to autologous DCs infected with viable Chlamydia, but not with DCs pulsed with inactivated chlamydial elementary bodies. MHC class Ia-restricted and non-class Ia-restricted responses were inhibited by DC treatment with a proteasomal inhibitor and an endoplasmic reticulum-Golgi transport inhibitor, suggesting that these T cells recognize a peptide Ag translocated to the host cell cytosol during infection that is processed via the classical class Ia Ag-processing pathway. Even though both restricted and nonrestricted CD8(+) T cells produced IFN-gamma in response to Chlamydia-infected fibroblasts, only the non-class Ia-restricted cells were lytic for these targets. The class Ia-restricted CTLs, however, were capable of cytolysis as measured by redirected killing. Collectively, these data demonstrate that both class Ia-restricted and non-classically restricted CD8(+) T cells are elicited in C. trachomatis-exposed individuals. Their role in host immunity remains to be elucidated. PMID- 14530354 TI - A dominant role of Toll-like receptor 4 in the signaling of apoptosis in bacteria faced macrophages. AB - Conserved bacterial components potently activate host immune cells through transmembrane Toll-like receptors (TLRs), which trigger a protective immune response but also may signal apoptosis. In this study, we investigated the roles of TLR2 and TLR4 as inducers of apoptosis in Yersinia enterocolitica-infected macrophages. Yersiniae suppress activation of the antiapoptotic NF-kappaB signaling pathway in host cells by inhibiting inhibitory kappaB kinase-beta. This leads to macrophage apoptosis under infection conditions. Experiments with mouse macrophages deficient for TLR2, TLR4, or both receptors showed that, although yersiniae could activate signaling through both TLR2 and TLR4, loss of TLR4 solely diminished Yersinia-induced apoptosis. This suggests implication of TLR4, but not of TLR2, as a proapoptotic signal transducer in Yersinia-conferred cell death. In the same manner, agonist-specific activation of TLR4 efficiently mediated macrophage apoptosis in the presence of the proteasome inhibitor MG-132, an effect that was less pronounced for activation through TLR2. Furthermore, the extended stimulation of overexpressed TLR4 elicited cellular death in epithelial cells. A dominant-negative mutant of Fas-associated death domain protein could suppress TLR4-mediated cell death, which indicates that TLR4 may signal apoptosis through a Fas-associated death domain protein-dependent pathway. Together, these data show that TLR4 could act as a potent inducer of apoptosis in macrophages that encounter a bacterial pathogen. PMID- 14530355 TI - Toll/IL-1 receptor domain-containing adaptor inducing IFN-beta (TRIF) associates with TNF receptor-associated factor 6 and TANK-binding kinase 1, and activates two distinct transcription factors, NF-kappa B and IFN-regulatory factor-3, in the Toll-like receptor signaling. AB - We previously reported a new Toll/IL-1R (TIR)-containing molecule, named TIR domain-containing adaptor inducing IFN-beta (TRIF). Although initial study indicated that TRIF possesses the ability to activate not only the NF-kappaB dependent but also the IFN-beta promoters, the molecular mechanisms of TRIF induced signaling are poorly understood. In this study, we investigated the signaling cascades through TRIF. TNF receptor-associated factor (TRAF)6 interacted with TRIF through the TRAF domain of TRAF6 and TRAF6-binding motifs found in the N-terminal portion of TRIF. Disruption of TRAF6-binding motifs of TRIF disabled it from associating with TRAF6, and resulted in a reduction in the TRIF-induced activation of the NF-kappaB-dependent but not IFN-beta promoter. TANK-binding kinase (TBK)-1, which was recently reported to be a kinase of IFN regulatory factor-3, which is an essential transcription factor for IFN-beta expression, also associated with the N-terminal region of TRIF. Moreover, the association between TRIF and TBK1 appeared to require the kinase activity of TBK1, as well as phosphorylation of TRIF. Because TRAF6 and TBK1 bind close the region of TRIF, it seems that TRAF6 physically prevents the association between TRIF and TBK1. Taken together, these results demonstrate that TRIF associates with TRAF6 and TBK1 independently, and activates two distinct transcription factors, NF-kappaB and IFN regulatory factor-3, respectively. PMID- 14530356 TI - Costimulatory molecule immune enhancement in a plasmid vaccine model is regulated in part through the Ig constant-like domain of CD80/86. AB - There is great interest in understanding the role of costimulatory molecules in immune activation. In both the influenza and HIV DNA immunization models, several groups have reported that coimmunization of mice with plasmids encoding immunogen and CD86, but not CD80, effectively boosts Ag-specific T cell activation. This difference in immune priming provided an opportunity to examine the functional importance of different regions of the B.7 molecules in immune activation. To examine this issue, we developed a series of chimeric CD80 and CD86 constructs as well as deletion mutants, and examined their immune activating potential in the DNA vaccine model. We demonstrate that the lack of an Ig constant-like region in the CD80 molecule is critically important to the enhanced immune activation observed. CD80 C-domain deletion mutants induce a highly inflammatory Ag-specific cellular response when administered as part of a plasmid vaccine. The data suggest that the constant-like domains, likely through intermolecular interactions, are critically important for immune regulation during costimulation and that engineered CD80/86 molecules represent more potent costimulatory molecules and may improve vaccine adjuvant efficacy. PMID- 14530357 TI - Toll-like receptor ligands modulate dendritic cells to augment cytomegalovirus- and HIV-1-specific T cell responses. AB - Optimal Ag targeting and activation of APCs, especially dendritic cells (DCs), are important in vaccine development. In this study, we report the effects of different Toll-like receptor (TLR)-binding compounds to enhance immune responses induced by human APCs, including CD123(+) plasmacytoid DCs (PDCs), CD11c(+) myeloid DCs (MDCs), monocytes, and B cells. PDCs, which express TLR7 and TLR9, responded to imidazoquinolines (imiquimod and R-848) and to CpG oligodeoxynucleotides stimulation, resulting in enhancement in expression of costimulatory molecules and induction of IFN-alpha and IL-12p70. In contrast, MDCs, which express TLR3, TLR4, and TLR7, responded to poly(I:C), LPS, and imidazoquinolines with phenotypic maturation and high production of IL-12 p70 without producing detectable IFN-alpha. Optimally TLR ligand-stimulated PDCs or MDCs exposed to CMV or HIV-1 Ags enhanced autologous CMV- and HIV-1-specific memory T cell responses as measured by effector cytokine production compared with TLR ligand-activated monocytes and B cells or unstimulated PDCs and MDCs. Together, these data show that targeting specific DC subsets using TLR ligands can enhance their ability to activate virus-specific T cells, providing information for the rational design of TLR ligands as adjuvants for vaccines or immune modulating therapy. PMID- 14530358 TI - Compromised host defense on Pseudomonas aeruginosa biofilms: characterization of neutrophil and biofilm interactions. AB - Pseudomonas aeruginosa is an opportunistic pathogen that forms biofilms on tissues and other surfaces. We characterized the interaction of purified human neutrophils with P. aeruginosa, growing in biofilms, with regard to morphology, oxygen consumption, phagocytosis, and degranulation. Scanning electron and confocal laser microscopy indicated that the neutrophils retained a round, unpolarized, unstimulated morphology when exposed to P. aeruginosa PAO1 biofilms. However, transmission electron microscopy demonstrated that neutrophils, although rounded on their dorsal side, were phagocytically active with moderate membrane rearrangement on their bacteria-adjacent surfaces. The settled neutrophils lacked pseudopodia, were impaired in motility, and were enveloped by a cloud of planktonic bacteria released from the biofilms. The oxygen consumption of the biofilm/neutrophil system increased 6- and 8-fold over that of the biofilm alone or unstimulated neutrophils in suspension, respectively. H(2)O(2) accumulation was transient, reaching a maximal measured value of 1 micro M. Following contact, stimulated degranulation was 20-40% (myeloperoxidase, beta-glucuronidase) and 40 80% (lactoferrin) of maximal when compared with formylmethionylleucylphenylalanine plus cytochalasin B stimulation. In summary, after neutrophils settle on P. aeruginosa biofilms, they become phagocytically engorged, partially degranulated, immobilized, and rounded. The settling also causes an increase in oxygen consumption of the system, apparently resulting from a combination of a bacterial respiration and escape response and the neutrophil respiratory burst but with little increase in the soluble concentration of H(2)O(2). Thus, host defense becomes compromised as biofilm bacteria escape while neutrophils remain immobilized with a diminished oxidative potential. PMID- 14530359 TI - Up-regulation of matrix metalloproteinase-9 in T lymphocytes of mammary tumor bearers: role of vascular endothelial growth factor. AB - Matrix metalloproteinase-9 (MMP-9), a matrix-degrading enzyme, is crucial in tumor invasion and metastasis and is implicated in leukocyte extravasation. In this report, we demonstrate that during growth of the D1-7,12 dimethylbenzanthracene-3 mammary tumor in BALB/c mice, there is progressive up regulation of MMP-9 in splenic T cells at both the transcriptional and translational levels. Our previous work has identified several factors produced by this tumor, including PGE(2), GM-CSF, and phosphatidyl serine; however, none of these agents induces increased production of MMP-9 by normal splenic T cells. Although not produced by the tumor, TNF-alpha and IL-6 are up-regulated in both macrophages and B cells in tumor-bearing mice. Exposure of normal T cells to these two cytokines, however, also fails to up-regulate MMP-9 production. Vascular endothelial growth factor (VEGF) is produced by many tumors, and we determined that the mammary tumor used in our studies expresses high levels of this angiogenic growth factor. Importantly, splenic T cells from tumor bearers constitutively produce increased amounts of VEGF, and treatment of normal T cells with VEGF results in up-regulated MMP-9 production. Of crucial importance is the finding that tumor-infiltrating T cells also produce high levels of VEGF and MMP 9. Our studies indicate that VEGF can act directly on T lymphocytes and that elevated VEGF levels may contribute to the aberrant MMP-9 secretion by mammary tumor bearers' T cells. PMID- 14530360 TI - Selective depletion of nonspecific T cells during the early stage of immune responses to infection. AB - Transient T cell depletion occurs before the development of an effective immune response to infection. In this study we show that most T cells, regardless of specificity, are induced to express early activation markers soon after infection with Listeria monocytogenes or lymphocytic choriomeningitis virus. Ag-specific T cells are further activated to display late activation markers and undergo extensive proliferation. As Ag-specific T cells begin to expand, nonspecific T cells are depleted en masse and exhibit no sign of further activation or proliferation before their depletion. This selective depletion of nonspecific T cells is due to in situ death via apoptosis, as visualized by confocal microscopy. Thus, early activation and subsequent depletion of nonspecific T cells are integral parts of the immune response to proinflammatory infections. These results have important implications for our understanding of early events in the development of a robust T cell response. PMID- 14530361 TI - Th1-biased tertiary lymphoid tissue supported by CXC chemokine ligand 13 producing stromal network in chronic lesions of autoimmune gastritis. AB - Secondary lymphoid tissue is developmentally programmed and characterized by well ordered compartmentalization of lymphocyte subsets and specialized stromal cells supporting the tissue architecture. By contrast, tertiary lymphoid tissue is defined as that induced in ectopic sites by inflammation, although its immunological role is largely unknown. In this study, we characterize the lymphoid tissue induced in the chronic lesion of murine autoimmune gastritis (AIG). Within the lymphoid cluster in the gastric mucosa, there is a clear segregation of T and B cells. Follicle-like B cell areas are always located on the luminal side of the mucosa, while T cells are located in the basal part. A typical lymphoid reticular network and follicular dendritic cells support the structure. Importantly, complement receptor 1(+) follicular dendritic cells within the follicle express a B cell homing chemokine, CXC chemokine ligand 13. The number and size of the clusters correlate with the age of the mice and the serum autoantibody titer, suggesting the functional importance of the clusters in local Ab production, although involvement of the autoantibody in the disease progression is still unclear. AIG gastric lesions are known to constitute a Th1 biased, memory T cell-dependent immunomicroenvironment. The expression pattern of cytokines, including lymphotoxin-beta, and chemokines in the AIG stomach is consistent with this observation. Taken together, these facts suggest that, during the chronic phase of autoimmunity, long-lasting lymphocyte infiltration probably induces a unique tertiary lymphoid tissue that has a function distinct from that of regional lymph nodes. These neolymphoid tissues may maintain the local self reactivity supporting the vicious cycle of Th1-type reaction as well as autoantibody production. PMID- 14530362 TI - Regulation of eotaxin gene expression by TNF-alpha and IL-4 through mRNA stabilization: involvement of the RNA-binding protein HuR. AB - During inflammatory responses, a major posttranscriptional regulation of early response and inflammatory gene expression occurs through modulation of mRNA turnover. We report that two potent inducers of the CC chemokine eotaxin, TNF alpha and IL-4, regulate its production in airway epithelial cells by increasing eotaxin mRNA stability. In experiments using the transcriptional inhibitor actinomycin D, eotaxin mRNA half-life was significantly prolonged by cell stimulation with TNF-alpha or IL-4, with the combination of the two cytokines being the most effective in extending the mRNA half-life. Involvement of the eotaxin 3' untranslated region in the mRNA-stabilizing effect was tested by transient transfection of a construct expressing a chimeric transcript carrying a serum-inducible beta-globin reporter linked to the eotaxin 3' untranslated region. The half-life of the chimeric mRNA was markedly increased in cells stimulated with TNF-alpha and IL-4. Evidence that the mRNA-stabilizing protein HuR participated in the cytokine effect was obtained: first, HuR presence in the cytoplasm, believed to be required for HuR-mediated mRNA stabilization, increased in both transformed (BEAS-2B cell line) and primary bronchial epithelial cells following treatment with TNF-alpha and IL-4. Second, endogenous eotaxin mRNA was found to bind to HuR in vivo, as detected by immunoprecipitation of HuR containing messenger ribonucleoprotein complexes followed by real-time RT-PCR analysis; such association increased after cell treatment with TNF-alpha and IL 4. Third, overexpression of HuR in BEAS-2B cells significantly increased the expression of eotaxin mRNA and protein. Our findings implicate mRNA stabilization in the cytokine-mediated increase in eotaxin expression and strongly suggest a role for HuR in this effect. PMID- 14530363 TI - Blockade of airway inflammation and hyperresponsiveness by HIV-TAT-dominant negative Ras. AB - We have reported previously that HIV-TAT-dominant negative (dn) Ras inhibits eosinophil adhesion to ICAM-1 after activation by IL-5 and eotaxin. In this study, we evaluated the role of Ras in Ag-induced airway inflammation and hyperresponsiveness by i.p. administration into mice of dnRas, which was fused to an HIV-TAT protein transduction domain (TAT-dnRas). Uptake of TAT-dnRas (t(1/2) = 12 h) was demonstrated in leukocytes after i.p. administration. OVA-sensitization significantly increased eosinophil and lymphocyte numbers in bronchoalveolar lavage fluid 24 h after final challenge. Treatment of animals with 3-10 mg/kg TAT dnRas blocked the migration of eosinophils from 464 +/- 91 x 10(3)/ml to 288 +/- 79 x 10(3)/ml with 3 mg/kg of TAT-dnRas (p < 0.05), and further decreased to 116 +/- 63 x 10(3)/ml after 10 mg/kg TAT-dnRas (p < 0.01). Histological examination demonstrated that inflammatory cell infiltration (largely eosinophils and mononuclear cells) and mucin production around the airways caused by OVA were blocked by TAT-dnRas. OVA challenge also caused airway hyperresponsiveness to methacholine, which was dose dependently blocked by treatment with TAT-dnRas. TAT dnRas also blocked Ag-induced IL-4 and IL-5, but not IFN-gamma, production in lung tissue. Intranasal administration of IL-5 caused eosinophil migration into the airway lumen, which was attenuated by pretreatment with TAT-dnRas. By contrast, TAT-green fluorescent protein or dnRas lacking the TAT protein transduction domain did not block airway inflammation, cytokine production, or airway hyperresponsiveness. We conclude that Ras mediates Th2 cytokine production, airway inflammation, and airway hyperresponsiveness in immune sensitized mice. PMID- 14530364 TI - Mast cells exert effects outside the central nervous system to influence experimental allergic encephalomyelitis disease course. AB - Previous studies using mast cell-deficient mice (W/W(v)) revealed that mast cells influence disease onset and severity of experimental allergic/autoimmune encephalomyelitis (EAE), the murine model for multiple sclerosis. The mast cell populations of these mice can be restored by transferring bone marrow-derived mast cells (BMMCs). Studies using the W/W(v) reconstitution model have lead to major advances in our understanding of mast cell roles in vivo. However, despite its common use, details regarding the sites and kinetics of mast cell repopulation have remained largely uncharacterized. In this study, we examined the kinetics and tissue distribution of green fluorescent protein(+) BMMCs in reconstituted W/W(v) mice to identify sites of mast cell influence in EAE. Reconstitution of naive animals with BMMCs does not restore mast cell populations to all organs, notably the brain, spinal cord, lymph nodes, and heart. Despite the absence of mast cells in the CNS, reconstituted mice exhibit an EAE disease course equivalent to that induced in wild-type mice. Mast cells are found adjacent to T cell-rich areas of the spleen and can migrate to the draining lymph node after disease induction. These data reveal that mast cells can act outside the CNS to influence EAE, perhaps by affecting the function of autoreactive lymphocytes. PMID- 14530365 TI - TNF-alpha induces phosphorylation of p47(phox) in human neutrophils: partial phosphorylation of p47phox is a common event of priming of human neutrophils by TNF-alpha and granulocyte-macrophage colony-stimulating factor. AB - Phosphorylation of p47(phox) is a key event in NADPH oxidase activation. We examined the ability of proinflammatory cytokines such as TNFalpha, IL-1, and G CSF to induce this process compared with GM-CSF. Only TNF-alpha and GM-CSF induced a clear p47(phox) phosphorylation. This phosphorylation was time dependent and reached its maximum at 20 min. Two-dimensional phosphopeptide mapping of p47(phox) phosphorylated in neutrophils primed with TNF-alpha revealed partial phosphorylation of p47(phox) on the same peptide as for GM-CSF. Neutrophil incubation with TNF-alpha and subsequent addition of the chemotactic peptide fMLP resulted in more intense phosphorylation of p47(phox) sites than with each reagent alone. A neutralizing Ab against the p55 TNF receptor, contrary to a neutralizing Ab against the p75 TNF receptor, inhibited TNF-alpha-induced p47(phox) phosphorylation. Neutrophil treatment with both TNF-alpha and GM-CSF resulted in more intense phosphorylation of the same p47(phox) peptide observed with each cytokine alone, suggesting that they engaged pathways converging on common serines. This additive effect was also obtained on the priming of NADPH oxidase activity. The use of protein kinase inhibitors pointed to the involvement of a protein tyrosine kinase, but not protein kinase C. These findings show that TNF-alpha, via its p55 receptor, induces a protein tyrosine kinase-dependent selective phosphorylation of p47(phox) on specific serines. The ability of TNF alpha and GM-CSF, two different cytokines with two different receptors to induce this specific p47(phox) phosphorylation, suggests that this event could be a common element of the priming of neutrophils by TNF-alpha and GM-CSF. PMID- 14530366 TI - Activation of group IV cytosolic phospholipase A2 in human eosinophils by phosphoinositide 3-kinase through a mitogen-activated protein kinase-independent pathway. AB - Activation of group IV cytosolic phospholipase A(2) (gIV-PLA(2)) is the essential first step in the synthesis of inflammatory eicosanoids and in integrin-mediated adhesion of leukocytes. Prior investigations have demonstrated that phosphorylation of gIV-PLA(2) results from activation of at least two isoforms of mitogen-activated protein kinase (MAPK). We investigated the potential role of phosphoinositide 3-kinase (PI3K) in the activation of gIV-PLA(2) and the hydrolysis of membrane phosphatidylcholine in fMLP-stimulated human blood eosinophils. Transduction into eosinophils of Deltap85, a dominant negative form of class IA PI3K adaptor subunit, fused to an HIV-TAT protein transduction domain (TAT-Deltap85) concentration dependently inhibited fMLP-stimulated phosphorylation of protein kinase B, a downstream target of PI3K. FMLP caused increased arachidonic acid (AA) release and secretion of leukotriene C(4) (LTC(4)). TAT-Deltap85 and LY294002, a PI3K inhibitor, blocked the phosphorylation of gIV-PLA(2) at Ser(505) caused by fMLP, thus inhibiting gIV PLA(2) hydrolysis and production of AA and LTC(4) in eosinophils. FMLP also caused extracellular signal-related kinases 1 and 2 and p38 MAPK phosphorylation in eosinophils; however, neither phosphorylation of extracellular signal-related kinases 1 and 2 nor p38 was inhibited by TAT-Deltap85 or LY294002. Inhibition of 1) p70 S6 kinase by rapamycin, 2) protein kinase B by Akt inhibitor, or 3) protein kinase C by Ro-31-8220, the potential downstream targets of PI3K for activation of gIV-PLA(2), had no effect on AA release or LTC(4) secretion caused by fMLP. We find that PI3K is required for gIV-PLA(2) activation and hydrolytic production of AA in activated eosinophils. Our data suggest that this essential PI3K independently activates gIV-PLA(2) through a pathway that does not involve MAPK. PMID- 14530367 TI - IL-8/CXCL8 and growth-related oncogene alpha/CXCL1 induce chondrocyte hypertrophic differentiation. AB - Foci of chondrocyte hypertrophy that commonly develop in osteoarthritic (OA) cartilage can promote dysregulated matrix repair and pathologic calcification in OA. The closely related chemokines IL-8/CXCL8 and growth-related oncogene alpha (GROalpha)/CXCL1 and their receptors are up-regulated in OA cartilage chondrocytes. Because these chemokines regulate leukocyte activation through p38 mitogen-activated protein kinase signaling, a pathway implicated in chondrocyte hypertrophic differentiation, we tested whether IL-8 and GROalpha promote chondrocyte hypertrophy. We observed that normal human and bovine primary articular chondrocytes expressed both IL-8Rs (CXCR1, CXCR2). IL-8 and the selective CXCR2 ligand GROalpha (10 ng/ml) induced tissue inhibitor of metalloproteinase-3 expression, markers of hypertrophy (type X collagen and MMP 13 expression, alkaline phosphatase activity), as well as matrix calcification. IL-8 and the selective CXCR2 ligand GROalpha also induced increased transamidation activity of chondrocyte transglutaminases (TGs), enzymes up regulated in chondrocyte hypertrophy that have the potential to modulate differentiation and calcification. Under these conditions, p38 mitogen-activated protein kinase pathway signaling mediated induction of both type X collagen and TG activity. Studies using mouse knee chondrocytes lacking one of the two known articular chondrocyte-expressed TG isoenzymes (TG2) demonstrated that TG2 was essential for murine GROalpha homologue KC-induced TG activity and critically mediated induction by KC of type X collagen, matrix metalloproteinase-13, alkaline phosphatase, and calcification. In conclusion, IL-8 and GROalpha induce articular chondrocyte hypertrophy and calcification through p38 and TG2. Our results suggest a novel linkage between inflammation and altered differentiation of articular chondrocytes. Furthermore, CXCR2 and TG2 may be sites for intervention in the pathogenesis of OA. PMID- 14530368 TI - Defective phagocytosis and clearance of Pseudomonas aeruginosa in the lung following bone marrow transplantation. AB - Bone marrow transplantation (BMT) is an important therapeutic option for a variety of malignant and nonmalignant disorders. Unfortunately, BMT recipients are at increased risk of infection, and in particular, pulmonary complications occur frequently. Although the risk of infection is greatest during the neutropenic period immediately following transplant, patients are still vulnerable to pulmonary infections even after neutrophil engraftment. We evaluated the risk of infection in this postengraftment period by using a well established mouse BMT model. Seven days after syngeneic BMT, B6D2F(1) mice are no longer neutropenic, and by 3 wk, they demonstrate complete reconstitution of the peripheral blood. However, these mice remain more susceptible throughout 8 wk to infection after intratracheal administration of Pseudomonas aeruginosa; increased mortality in the P. aeruginosa-infected BMT mice correlates with increased bacterial burden in the lungs as well as increased systemic dissemination. This heightened susceptibility to infection was not secondary to a defect in inflammatory cell recruitment to the lung. The inability to clear P. aeruginosa in the lung correlated with reduced phagocytosis of the bacteria by alveolar macrophages (AMs), but not neutrophils, decreased production of TNF-alpha by AMs, and decreased levels of TNF-alpha and IFN-gamma in the bronchoalveolar lavage fluid following infection. Expression of the beta(2) integrins CD11a and CD11c was reduced on AMs from BMT mice compared with wild-type mice. Thus, despite restoration of peripheral blood count, phagocytic defects in the AMs of BMT mice persist and may contribute to the increased risk of infection seen in the postengraftment period. PMID- 14530369 TI - The hemopoietic Rho/Rac guanine nucleotide exchange factor Vav1 regulates N formyl-methionyl-leucyl-phenylalanine-activated neutrophil functions. AB - Vav1 is a hemopoietic-specific Rho/Rac guanine nucleotide exchange factor that plays a prominent role in responses to multisubunit immune recognition receptors in lymphoid cells, but its contribution to regulation of neutrophil functions is unknown. Activated Rho family GTPases are critical participants in neutrophil signaling cascades initiated by binding of FMLP and other chemoattractants to their cognate G protein-coupled receptors. Therefore, we investigated whether Vav1 regulates chemoattractant-induced responses in neutrophils. We found that superoxide production elicited by FMLP in Vav1(-/-) murine neutrophils isolated from either bone marrow or from peritoneal exudates was substantially reduced compared with that of wild type. Filamentous actin generation in FMLP-stimulated Vav1(-/-) neutrophils was also markedly reduced, whereas it was normal in response to IL-8 or leukotriene B(4). FMLP induced tyrosine phosphorylation of Vav1, whereas IL-8 or leukotriene B(4) did not, correlating with the requirement for Vav1 in chemoattractant-stimulated filamentous actin generation. Neutrophil motility in vitro and neutrophil mobilization into peripheral blood in vivo elicited by FMLP were both decreased in Vav1(-/-) mice. Hence, this study defines a new role for Vav1 in regulating granulocytic leukocytes as well as linking Vav1 to specific cellular responses downstream of a seven transmembrane domain receptor. PMID- 14530370 TI - HLA-DRB1*0402 (DW10) transgene protects collagen-induced arthritis-susceptible H2Aq and DRB1*0401 (DW4) transgenic mice from arthritis. AB - To investigate the role of HLA-DR4 in predisposition to arthritis, we generated transgenic mice carrying DRB1*0401 and DRB1*0402 genes. We have previously shown that DRB1*0401 molecule renders B10.RQB3 (H2A(q)) mice susceptible to porcine and human type II collagen-induced arthritis. We report that the introduction of DRB1*0402 transgene does not lead to development of arthritis in mice when they are immunized with porcine and human type II collagen. In addition, DRB1*0402 protects B10.RQB3 mice against developing arthritis with bovine type II collagen. These data show that DRB1 can modulate the disease mediated by A(q). In vivo depletion of DRB1*0402 did not lead to induction of collagen-induced arthritis in transgenic mice. In vitro cytokine analysis shows that mice protected from collagen-induced arthritis produce lower amounts of Th1 and higher levels of Th2 type cytokines upon immunization with type II collagen. Protection of mice was also related to higher apoptosis in DW10 mice as indicated by higher amounts of BclII in response to type II collagen. On the basis of our observations in HLA transgenic mice, we hypothesize that DRB1 polymorphism can modulate disease by shaping the T cell repertoire in thymus and select autoreactive T cells. PMID- 14530371 TI - Repeated alpha-galactosylceramide administration results in expansion of NK T cells and alleviates inflammatory dermatitis in MRL-lpr/lpr mice. AB - NK T (NKT) cells expressing the invariant Valpha14-Jalpha18 TCR alpha-chain recognize glycolipid Ags such as alpha-galactosylceramide (alpha-GalCer) presented by the MHC class I-like molecule CD1d. Upon activation by alpha-GalCer, invariant NKT cells secrete multiple cytokines and confer protection in certain immune-mediated disorders. Here we have investigated the role of NKT cells in the development of inflammatory dermatitis in MRL-lpr/lpr mice, which shares features with lupus in humans. Our results show that the numbers Sand functions of NKT (TCRbeta(+)CD1d/alpha-GalCer tetramer(+)) cells, particularly of the NK1.1(-) subset, are reduced in MRL-lpr/lpr mice compared with MRL-fas/fas and/or nonautoimmune C3H/Hej and BALB/c mice. Repeated treatments with alpha-GalCer result in the expansion of NKT cells and alleviate dermatitis in MRL-lpr/lpr mice. Our results indicate that NKT cell deficiency can be corrected by repeated alpha-GalCer treatment and that NKT cells may play a protective role in inflammatory dermatitis of lupus-prone mice. PMID- 14530372 TI - IL-7 stimulates T cell renewal without increasing viral replication in simian immunodeficiency virus-infected macaques. AB - The main failure of antiretroviral therapy is the lack of restoration of HIV specific CD4(+) T cells. IL-7, which has been shown to be a crucial cytokine for thymopoiesis, has been envisaged as an additive therapeutic strategy. However, in vitro studies suggest that IL-7 might sustain HIV replication in thymocytes and T lymphocytes. Therefore, in the present study, we evaluated the effect of IL-7 on both T cell renewal and viral load in SIVmac-infected young macaques in the absence of antiretroviral therapy. This evaluation was conducted during the asymptomatic phase in view of a potential treatment of HIV patients. We show that IL-7 induces both a central renewal and a peripheral expansion of T lymphocytes associated with cell activation. No alarming modulation of the other hemopoietic cells was observed. No increase in the viral load was shown in blood or lymph nodes. These data strengthen the rationale for the use of IL-7 as an efficient immunotherapy in AIDS. PMID- 14530373 TI - Specific recruitment of antigen-presenting cells by chemerin, a novel processed ligand from human inflammatory fluids. AB - Dendritic cells (DCs) and macrophages are professional antigen-presenting cells (APCs) that play key roles in both innate and adaptive immunity. ChemR23 is an orphan G protein-coupled receptor related to chemokine receptors, which is expressed specifically in these cell types. Here we present the characterization of chemerin, a novel chemoattractant protein, which acts through ChemR23 and is abundant in a diverse set of human inflammatory fluids. Chemerin is secreted as a precursor of low biological activity, which upon proteolytic cleavage of its COOH terminal domain, is converted into a potent and highly specific agonist of ChemR23, the chemerin receptor. Activation of chemerin receptor results in intracellular calcium release, inhibition of cAMP accumulation, and phosphorylation of p42-p44 MAP kinases, through the Gi class of heterotrimeric G proteins. Chemerin is structurally and evolutionary related to the cathelicidin precursors (antibacterial peptides), cystatins (cysteine protease inhibitors), and kininogens. Chemerin was shown to promote calcium mobilization and chemotaxis of immature DCs and macrophages in a ChemR23-dependent manner. Therefore, chemerin appears as a potent chemoattractant protein of a novel class, which requires proteolytic activation and is specific for APCs. PMID- 14530374 TI - Antiadhesive role of apical decay-accelerating factor (CD55) in human neutrophil transmigration across mucosal epithelia. AB - Neutrophil migration across mucosal epithelium during inflammatory episodes involves the precise orchestration of a number a cell surface molecules and signaling pathways. After successful migration to the apical epithelial surface, apically localized epithelial proteins may serve to retain PMN at the lumenal surface. At present, identification of apical epithelial ligands and their PMN counter-receptors remain elusive. Therefore, to define the existence of apical epithelial cell surface proteins involved in PMN-epithelial interactions, we screened a panel of antibodies directed against epithelial plasma membranes. This strategy identified one antibody (OE-1) that both localized to the apical cell membrane and significantly inhibited PMN transmigration across epithelial monolayers. Microsequence analysis revealed that OE-1 recognized human decay accelerating factor (DAF, CD55). DAF is a highly glycosylated, 70-80-kD, glycosyl phosphatidyinositol-linked protein that functions predominantly as an inhibitor of autologous complement lysis. DAF suppression experiments using antisense oligonucleotides or RNA interference revealed that DAF may function as an antiadhesive molecule promoting the release of PMN from the lumenal surface after transmigration. Similarly, peptides corresponding to the antigen recognition domain of OE-1 resulted in accumulation of PMN on the apical epithelial surface. The elucidation of DAF as an apical epithelial ligand for PMN provides a target for novel anti-inflammatory therapies directed at quelling unwanted inflammatory episodes. PMID- 14530375 TI - Route of immunization with peptide-pulsed dendritic cells controls the distribution of memory and effector T cells in lymphoid tissues and determines the pattern of regional tumor control. AB - We have established that the route of immunization with peptide-pulsed, activated DC leads to memory CD8+ T cells with distinct distributions in lymphoid tissues, which determines the ability to control tumors growing in different body sites. Both intravenous (i.v.) and subcutaneous (s.c.) immunization induced memory T cells in spleen and control of metastatic-like lung tumors. s.c. immunization also induced memory T cells in lymph nodes (LNs), imparting protection against subcutaneously growing tumors. In contrast, i.v. immunization-induced memory was restricted to spleen and failed to impart protective immunity against subcutaneously growing tumors. Memory cell distribution and tumor control were both linked to injection route-dependent localization of DCs in lymphoid compartments. Using peripheral LN-ablated mice, these LNs were shown to be essential for control of subcutaneously growing tumors but not lung metastases; in contrast, using immunized asplenic mice, we found that the spleen is necessary and sufficient for control of lung tumors, but unnecessary for control of subcutaneously growing tumors. These data demonstrate the existence of a previously undescribed population of splenic-resident memory CD8 T cells that are essential for the control of lung metastases. Thus, regional immunity based on memory T cell residence patterns is an important factor in DC-based tumor immunotherapy. PMID- 14530376 TI - Constitutive cytokine mRNAs mark natural killer (NK) and NK T cells poised for rapid effector function. AB - Natural killer (NK) and NK T cells are tissue lymphocytes that secrete cytokines rapidly upon stimulation. Here, we show that these cells maintain distinct patterns of constitutive cytokine mRNAs. Unlike conventional T cells, NK T cells activate interleukin (IL)-4 and interferon (IFN)-gamma transcription during thymic development and populate the periphery with both cytokine loci previously modified by histone acetylation. Similarly, NK cells transcribe and modify the IFN-gamma gene, but not IL-4, during developmental maturation in the bone marrow. Lineage-specific patterns of cytokine transcripts predate infection and suggest evolutionary selection for invariant but distinct types of effector responses among the earliest responding lymphocytes. PMID- 14530377 TI - Targeting platelet-leukocyte interactions: identification of the integrin Mac-1 binding site for the platelet counter receptor glycoprotein Ibalpha. AB - The firm adhesion and transplatelet migration of leukocytes on vascular thrombus are dependent on the interaction of the leukocyte integrin Mac-1 (alphaMbeta2, CD11b/CD18) and the platelet counter receptor glycoprotein (GP) Ibalpha. Previous studies have established a central role for the I domain, a stretch of approximately 200 amino acids within the alphaM subunit, in the binding of GP Ibalpha. This study was undertaken to establish the molecular basis of GP Ibalpha recognition by alphaMbeta2. The P201-K217 sequence, which spans an exposed loop and amphipathic alpha4 helix in the three-dimensional structure of the alphaMI domain, was identified as the binding site for GP Ibalpha. Mutant cell lines in which the alphaMI domain segments P201-G207 and R208-K217 were switched to the homologous, but non-GP Ibalpha binding, alphaL domain segments failed to support adhesion to GP Ibalpha. Mutation of amino acid residues within P201-K217, H210 A212, T213-I215, and R216-K217 resulted in the loss of the binding function of the recombinant alphaMI domains to GP Ibalpha. Synthetic peptides duplicating the P201-K217, but not scrambled versions, directly bound GP Ibalpha and inhibited alphaMbeta2-dependent adhesion to GP Ibalpha and adherent platelets. Finally, grafting critical amino acids within the P201-K217 sequence onto alphaL, converted alphaLbeta2 into a GP Ibalpha binding integrin. Thus, the P201-K217 sequence within the alphaMI domain is necessary and sufficient for GP Ibalpha binding. These observations provide a molecular target for disrupting leukocyte platelet complexes that promote vascular inflammation in thrombosis, atherosclerosis, and angioplasty-related restenosis. PMID- 14530378 TI - Effective destruction of Fas-deficient insulin-producing beta cells in type 1 diabetes. AB - In type 1 diabetes, autoimmune T cells cause destruction of pancreatic beta cells by largely unknown mechanism. Previous analyses have shown that beta cell destruction is delayed but can occur in perforin-deficient nonobese diabetic (NOD) mice and that Fas-deficient NOD mice do not develop diabetes. However, because of possible pleiotropic functions of Fas, it was not clear whether the Fas receptor was an essential mediator of beta cell death in type 1 diabetes. To directly test this hypothesis, we have generated a beta cell-specific knockout of the Fas gene in a transgenic model of type 1 autoimmune diabetes in which CD4+ T cells with a transgenic TCR specific for influenza hemagglutinin (HA) are causing diabetes in mice that express HA under control of the rat insulin promoter. Here we show that the Fas-deficient mice develop autoimmune diabetes with slightly accelerated kinetics indicating that Fas-dependent apoptosis of beta cells is a dispensable mode of cell death in this disease. PMID- 14530379 TI - The strength of persistent antigenic stimulation modulates adaptive tolerance in peripheral CD4+ T cells. AB - The quantitative adaptation of receptor thresholds allows cells to tailor their responses to changes in ambient ligand concentration in many biological systems. Such a cell-intrinsic calibration of T cell receptor (TCR) sensitivity could be involved in regulating responses to autoantigens, but this has never been demonstrated for peripheral T cells. We examined the ability of monoclonal naive T cells to modulate their responsiveness differentially after exposure to fourfold different levels of persistent antigen stimulation in vivo. T cells expanded and entered a tolerant state with different kinetics in response to the two levels of stimulation, but eventually adjusted to a similar slow rate of turnover. In vivo restimulation revealed a greater impairment in the proliferative ability of T cells resident in a higher antigen presentation environment. We also observed subtle differences in TCR signaling and in vitro cytokine production consistent with differential adaptation. Unexpectedly, the system failed to similarly compensate to the persistent stimulus in vivo at the level of CD69 expression and actin polymerization. This greater responsiveness of T cells residing in a host with a lower level of antigen presentation allows us to demonstrate for the first time an intrinsic tuning process in mature T lymphocytes, albeit one more complex than current theories predict. PMID- 14530380 TI - Heparan sulfate regulates amyloid precursor protein processing by BACE1, the Alzheimer's beta-secretase. AB - Cleavage of amyloid precursor protein (APP) by the Alzheimer's beta-secretase (BACE1) is a key step in generating amyloid beta-peptide, the main component of amyloid plaques. Here we report evidence that heparan sulfate (HS) interacts with beta-site APP-cleaving enzyme (BACE) 1 and regulates its cleavage of APP. We show that HS and heparin interact directly with BACE1 and inhibit in vitro processing of peptide and APP substrates. Inhibitory activity is dependent on saccharide size and specific structural characteristics, and the mechanism of action involves blocking access of substrate to the active site. In cellular assays, HS specifically inhibits BACE1 cleavage of APP but not alternative cleavage by alpha secretase. Endogenous HS immunoprecipitates with BACE1 and colocalizes with BACE1 in the Golgi complex and at the cell surface, two of its putative sites of action. Furthermore, inhibition of cellular HS synthesis results in enhanced BACE1 activity. Our findings identify HS as a natural regulator of BACE1 and suggest a novel mechanism for control of APP processing. PMID- 14530381 TI - MAL regulates clathrin-mediated endocytosis at the apical surface of Madin-Darby canine kidney cells. AB - MAL is an integral protein component of the machinery for apical transport in epithelial Madin-Darby canine kidney (MDCK) cells. To maintain its distribution, MAL cycles continuously between the plasma membrane and the Golgi complex. The clathrin-mediated route for apical internalization is known to differ from that at the basolateral surface. Herein, we report that MAL depends on the clathrin pathway for apical internalization. Apically internalized polymeric Ig receptor (pIgR), which uses clathrin for endocytosis, colocalized with internalized MAL in the same apical vesicles. Time-lapse confocal microscopic analysis revealed cotransport of pIgR and MAL in the same endocytic structures. Immunoelectron microscopic analysis evidenced colabeling of MAL with apically labeled pIgR in pits and clathrin-coated vesicles. Apical internalization of pIgR was abrogated in cells with reduced levels of MAL, whereas this did not occur either with its basolateral entry or the apical internalization of glycosylphosphatidylinositol anchored proteins, which does not involve clathrin. Therefore, MAL is critical for efficient clathrin-mediated endocytosis at the apical surface in MDCK cells. PMID- 14530383 TI - H2AX regulates meiotic telomere clustering. AB - The histone H2A variant H2AX is phosphorylated in response to DNA double-strand breaks originating from diverse origins, including dysfunctional telomeres. Here, we show that normal mitotic telomere maintenance does not require H2AX. Moreover, H2AX is dispensable for the chromosome fusions arising from either critically shortened or deprotected telomeres. However, H2AX has an essential role in controlling the proper topological distribution of telomeres during meiotic prophase I. Our results suggest that H2AX is a downstream effector of the ataxia telangiectasia-mutated kinase in controlling telomere movement during meiosis. PMID- 14530382 TI - Actin cytoskeleton remodeling during early Drosophila furrow formation requires recycling endosomal components Nuclear-fallout and Rab11. AB - Cytokinesis requires a dramatic remodeling of the cortical cytoskeleton as well as membrane addition. The Drosophila pericentrosomal protein, Nuclear-fallout (Nuf), provides a link between these two processes. In nuf-derived embryos, actin remodeling and membrane recruitment during the initial stages of metaphase and cellular furrow formation are disrupted. Nuf is a homologue of arfophilin-2, an ADP ribosylation factor effector that binds Rab11 and influences recycling endosome (RE) organization. Here, we show that Nuf is an important component of the RE, and that these phenotypes are a consequence of Nuf activities at the RE. Nuf exhibits extensive colocalization with Rab11, a key RE component. GST pull downs and the presence of a conserved Rab11-binding domain in Nuf demonstrate that Nuf and Rab11 physically associate. In addition, Nuf and Rab11 are mutually required for their localization to the RE. Embryos with reduced levels of Rab11 produce membrane recruitment and actin remodeling defects strikingly similar to nuf-derived embryos. These analyses support a common role for Nuf and Rab11 at the RE in membrane trafficking and actin remodeling during the initial stages of furrow formation. PMID- 14530384 TI - Ligand crowding at a nascent signal sequence. AB - We have systematically analyzed the molecular environment of the signal sequence of a growing secretory protein from Escherichia coli using a stage- and site specific cross-linking approach. Immediately after emerging from the ribosome, the signal sequence of pOmpA is accessible to Ffh, the protein component of the bacterial signal recognition particle, and to SecA, but it remains attached to the surface of the ribosome via protein L23. These contacts are lost upon further growth of the nascent chain, which brings the signal sequence into sole proximity to the chaperone Trigger factor (TF). In its absence, nascent pOmpA shows extended contacts with L23, and even long chains interact in these conditions proficiently with Ffh. Our results suggest that upon emergence from the ribosome, the signal sequence of an E. coli secretory protein gradually becomes sequestered by TF. Although TF thereby might control the accessibility of pOmpA's signal sequence to Ffh and SecA, it does not influence interaction of pOmpA with SecB. PMID- 14530385 TI - Inhibition of NF-kappaB by ZAS3, a zinc-finger protein that also binds to the kappaB motif. AB - The ZAS proteins are large zinc-finger transcriptional proteins implicated in growth, signal transduction, and lymphoid development. Recombinant ZAS fusion proteins containing one of the two DNA-binding domains have been shown to bind specifically to the kappaB motif, but the endogenous ZAS proteins or their physiological functions are largely unknown. The kappaB motif, GGGACTTTCC, is a gene regulatory element found in promoters and enhancers of genes involved in immunity, inflammation, and growth. The Rel family of NF-kappaB, predominantly p65.p50 and p50.p50, are transcription factors well known for inducing gene expression by means of interaction with the kappaB motif during acute-phase responses. A functional link between ZAS and NF-kappaB, two distinct families of kappaB-binding proteins, stems from our previous in vitro studies that show that a representative member, ZAS3, associates with TRAF2, an adaptor molecule in tumor necrosis factor signaling, to inhibit NF-kappaB activation. Biochemical and genetic evidence presented herein shows that ZAS3 encodes major kappaB-binding proteins in B lymphocytes, and that NF-kappaB is constitutively activated in ZAS3 deficient B cells. The data suggest that ZAS3 plays crucial functions in maintaining cellular homeostasis, at least in part by inhibiting NF-kappaB by means of three mechanisms: inhibition of nuclear translocation of p65, competition for kappaB gene regulatory elements, and repression of target gene transcription. PMID- 14530386 TI - Nuclear localization of 5-lipoxygenase as a determinant of leukotriene B4 synthetic capacity. AB - The enzyme 5-lipoxygenase (5-LO) initiates the synthesis of leukotrienes from arachidonic acid. In resting cells, 5-LO can accumulate in either the cytoplasm or the nucleoplasm and, upon cell stimulation, translocates to membranes to initiate leukotriene synthesis. Here, we used mutants of 5-LO with altered subcellular localization to assess the role that nuclear positioning plays in determining leukotriene B4 (LTB4) synthesis. Mutation of either a nuclear localization sequence or a phosphorylation site reduced LTB4 synthesis by 60%, in parallel with reduced nuclear localization of 5-LO. Mutation of both sites together or mutation of all three nuclear localization sequences on 5-LO inhibited LTB4 synthesis by 90% and abolished nuclear localization. Reduced LTB4 generation in mutants could not be attributed to differences in 5-LO amount, enzymatic activity, or membrane association. Instead, 5-LO within the nucleus acts at a different site, the nuclear envelope, than does cytosolic 5-LO, which acts at cytoplasmic and perinuclear membranes. The significance of this difference was suggested by evidence that exogenously derived arachidonic acid colocalized with activated nuclear 5-LO. These results unequivocally demonstrate that the positioning of 5-LO within the nucleus of resting cells is a powerful determinant of the capacity to generate LTB4 upon subsequent activation. PMID- 14530387 TI - Requirement for the TIE family of receptor tyrosine kinases in adult but not fetal hematopoiesis. AB - In mammals, the continuous production of hematopoietic cells (HCs) is sustained by a small number of hematopoietic stem cells (HSCs) residing in the bone marrow. Early HSC activity arises in the aorta-gonad mesonephros region, within cells localized to the ventral floor of the major blood vessels, suggesting that the first HSCs may be derived from cells capable of giving rise to the hematopoietic system and to the endothelial cells of the vasculature. TIE1 (TIE) and TIE2 (TEK) are related receptor tyrosine kinases with an embryonic expression pattern in endothelial cells, their precursors, and HCs, suggestive of a role in the divergence and function of both lineages. Indeed, gene targeting approaches have shown that TIE1, TIE2, and ligands for TIE2, the angiopoietins, are essential for vascular development and maintenance. To explore possible roles for these receptors in HCs, we have examined the ability of embryonic cells lacking both TIE1 and TIE2 to contribute to developmental and adult hematopoiesis by generating chimeric animals between normal embryonic cells and cells lacking these receptors. We show here that TIE receptors are not required for differentiation and proliferation of definitive hematopoietic lineages in the embryo and fetus; surprisingly, however, these receptors are specifically required during postnatal bone marrow hematopoiesis. PMID- 14530388 TI - Structure and function of the feed-forward loop network motif. AB - Engineered systems are often built of recurring circuit modules that carry out key functions. Transcription networks that regulate the responses of living cells were recently found to obey similar principles: they contain several biochemical wiring patterns, termed network motifs, which recur throughout the network. One of these motifs is the feed-forward loop (FFL). The FFL, a three-gene pattern, is composed of two input transcription factors, one of which regulates the other, both jointly regulating a target gene. The FFL has eight possible structural types, because each of the three interactions in the FFL can be activating or repressing. Here, we theoretically analyze the functions of these eight structural types. We find that four of the FFL types, termed incoherent FFLs, act as sign-sensitive accelerators: they speed up the response time of the target gene expression following stimulus steps in one direction (e.g., off to on) but not in the other direction (on to off). The other four types, coherent FFLs, act as sign-sensitive delays. We find that some FFL types appear in transcription network databases much more frequently than others. In some cases, the rare FFL types have reduced functionality (responding to only one of their two input stimuli), which may partially explain why they are selected against. Additional features, such as pulse generation and cooperativity, are discussed. This study defines the function of one of the most significant recurring circuit elements in transcription networks. PMID- 14530389 TI - Juvenile hormone acid methyltransferase: a key regulatory enzyme for insect metamorphosis. AB - Juvenile hormone (JH) acid methyltransferase (JHAMT) is an enzyme that converts JH acids or inactive precursors of JHs to active JHs at the final step of JH biosynthesis pathway in insects. By fluorescent mRNA differential display, we have cloned a cDNA encoding JHAMT from the corpora allata (CA) of the silkworm, Bombyx mori (BmJHAMT). The BmJHAMT cDNA encodes an ORF of 278 aa with a calculated molecular mass of 32,544 Da. The predicted amino acid sequence contains a conserved S-adenosyl-l-methionine (SAM) binding motif found in the family of SAM-dependent methyltransferases. Purified N-terminal 6xHis-tagged recombinant BmJHAMT protein expressed in Escherichia coli catalyzed conversion of farnesoic acid and JH acids I, II, and III to their cognate methyl esters in the presence of SAM, confirming that this cDNA encodes a functional JHAMT. Putative orthologs, DmJHAMT and AgJHAMT, were identified from the genome sequence of the fruit fly Drosophila melanogaster, and a malaria vector, Anopheles gambiae, respectively. Northern blot and quantitative RT-PCR analyses revealed that the BmJHAMT gene was expressed specifically in the CA throughout the third and fourth instar. At the beginning of the last (fifth) instar, the expression level of BmJHAMT declined rapidly and became undetectable by day 4 and remained so until pupation. Correlation of the BmJHAMT gene expression and the JH biosynthetic activity in the CA suggests that the transcriptional suppression of the BmJHAMT gene is crucial for the termination of JH biosynthesis in the CA, which is a prerequisite for the initiation of metamorphosis. PMID- 14530390 TI - The energetic cost of domain reorientation in maltose-binding protein as studied by NMR and fluorescence spectroscopy. AB - Maltose-binding protein (MBP) is a two-domain protein that undergoes a ligand mediated conformational rearrangement from an "open" to a "closed" structure on binding to maltooligosaccharides. To characterize the energy landscape associated with this transition, we have generated five variants of MBP with mutations located in the hinge region of the molecule. Residual dipolar couplings, measured in the presence of a weak alignment medium, have been used to establish that the average structures of the mutant proteins are related to each other by domain rotation about an invariant axis, with the rotation angle varying from 5 degrees to 28 degrees. Additionally, the domain orientations observed in the wild-type apo and ligand-bound (maltose, maltotriose, etc.) structures are related through a rotation of 35 degrees about the same axis. Remarkably, the free energy of unfolding, measured by equilibrium denaturation experiments and monitored by fluorescence spectroscopy, shows a linear correlation with the rotation angle, with the stability of the (apo)protein decreasing with domain closure by 212 +/- 16 cal mol-1 per degree of rotation. The apparent binding energy for maltose also shows a similar correlation with the interdomain angle, suggesting that the mutations, as they relate to binding, affect predominantly the ligand-free structure. The linearity of the energy change is interpreted in terms of an increase in the extent of hydrophobic surface that becomes solvent accessible on closure. The combination of structural, stability, and binding data allows separation of the energetics of domain reorientation from ligand binding. This work presents a near quantitative structure-energy-binding relationship for a series of mutants of MBP, illustrating the power of combined studies involving protein engineering and solution NMR spectroscopy. PMID- 14530391 TI - PPARgamma coactivator 1beta/ERR ligand 1 is an ERR protein ligand, whose expression induces a high-energy expenditure and antagonizes obesity. AB - A well balanced body energy budget controlled by limitation of calorie uptake and/or increment of energy expenditure, which is typically achieved by proper physical exercise, is most effective against obesity and diabetes mellitus. Recently, peroxisome proliferator-activated receptor (PPAR) gamma, a member of the nuclear receptor, and its cofactors have been shown to be involved in lipid metabolism and in the control of energy expenditure. Here we show that PPARgamma coactivator 1 (PGC-1) beta functions as ERRL1 (for ERR ligand 1), which can bind and activate orphan ERRs (estrogen receptor-related receptors) in vitro. Consistently, PGC-1beta/ERRL1 transgenic mice exhibit increased expression of the medium-chain acyl CoA dehydrogenase, a known ERR target and a pivotal enzyme of mitochondrial beta-oxidation in skeletal muscle. As a result, the PGC-1beta/ERRL1 mice show a state similar to an athlete; namely, the mice are hyperphagic and of elevated energy expenditure and are resistant to obesity induced by a high-fat diet or by a genetic abnormality. These results demonstrate that PGC-1beta/ERRL1 can function as a protein ligand of ERR, and that its level contributes to the control of energy balance in vivo, and provide a strategy for developing novel antiobesity drugs. PMID- 14530392 TI - The HLA-DQ2 gene dose effect in celiac disease is directly related to the magnitude and breadth of gluten-specific T cell responses. AB - In patients with celiac disease, inflammatory T cell responses to HLA-DQ2-bound gluten peptides are thought to cause disease. Two types of HLA-DQ2 molecules exist, termed HLA-DQ2.5 and HLA-DQ2.2. Whereas HLA-DQ2.5 predisposes to celiac disease, HLA-DQ2.2 does not. We now provide evidence that the disease-associated HLA-DQ2.5 molecule presents a large repertoire of gluten peptides, whereas the non-disease-associated HLA-DQ2.2 molecule can present only a subset of these. Moreover, gluten presentation by HLA-DQ2 homozygous antigen-presenting cells was superior to presentation by HLA-DQ2/non-DQ2 heterozygous antigen-presenting cells in terms of T cell proliferation and cytokine secretion. Gluten presentation by HLA-DQ2.5/2.2 heterozygous antigen-presenting cells induced intermediate T cell stimulation. These results correlated with peptide binding to the antigen presenting cells. Finally, we demonstrate that HLA-DQ trans dimers formed in HLA DQ2.5/2.2 heterozygous individuals have properties identical with HLA-DQ2.5 dimers. Our findings explain the strongly increased risk of disease development for HLA-DQ2.5 homozygous and HLA-DQ2.2/2.5 heterozygous individuals, and they are indicative of a quantitative model for disease development, where HLA-DQ expression and the available number of T cell-stimulatory gluten peptides are critical limiting factors. This model may have important implications for disease prevention. PMID- 14530394 TI - Manduca sexta recognition and resistance among allopolyploid Nicotiana host plants. AB - Allopolyploid speciation occurs instantly when the genomes of different species combine to produce self-fertile offspring and has played a central role in the evolution of higher plants, but its consequences for adaptive responses are unknown. We compare herbivore-recognition and -resistance responses of the diploid species and putative ancestral parent Nicotiana attenuata with those of the two derived allopolyploid species Nicotiana clevelandii and Nicotiana bigelovii. Manduca sexta larvae attack all three species, and in N. attenuata attack is recognized when larval oral secretions are introduced to wounds during feeding, resulting in a jasmonate burst, a systemic amplification of trypsin inhibitor accumulation, and a release of volatile organic compounds, which function as a coordinated defense response that slows caterpillar growth and increases the probability of their being attacked. Most aspects of this recognition response are retained with modifications in one allotetraploid (N. bigelovii) but lost in the other (N. clevelandii). Differences between diploid and tetraploid species were apparent in delays (maximum 1 and 0.5 h, respectively) in the jasmonate burst, the elicitation of trypsin inhibitors and release of volatile organic compounds, and the constitutive levels of nicotine, trypsin inhibitors, diterpene glycosides, rutin, and caffeoylputrescine in the leaves. Resistance to M. sexta larvae attack was most strongly associated with diterpene glycosides, which were higher in the diploid than in the two allotetraploid species. Because M. sexta elicitors differentially regulate a large proportion of the N. attenuata transcriptome, we propose that these species are suited for the study of the evolution of adaptive responses requiring trans activation mechanisms. PMID- 14530393 TI - Y box-binding protein 1 induces resistance to oncogenic transformation by the phosphatidylinositol 3-kinase pathway. AB - Y box-binding protein 1 (YB-1) is a multifunctional protein that can act as a regulator of transcription and of translation. In chicken embryo fibroblasts transformed by the oncoproteins P3k (phosphatidylinositol 3-kinase) or Akt, YB-1 is transcriptionally down-regulated. Expression of YB-1 from a retroviral vector induces a strong cellular resistance to transformation by P3k or Akt but does not affect sensitivity to transformation by other oncoproteins, such as Src, Jun, or Qin. The YB-1-expressing cells assume a tightly adherent, flat phenotype, with YB 1 localized in the cytoplasm, and show a greatly reduced saturation density. Both cap-dependent and cap-independent translation is inhibited in these cells, but the activity of Akt remains unaffected, suggesting that YB-1 functions downstream of Akt. A YB-1 protein with a loss-of-function mutation in the RNA-binding motif no longer binds to the mRNA cap structure, is localized in the cell nucleus, does not induce the flat cellular phenotype, and fails to interfere with P3k- or Akt induced oncogenic transformation. This mutant also does not inhibit cap-dependent or cap-independent translation. These results suggest that YB-1 acts like a rapamycin mimic, inhibiting translational events that are required in phosphatidylinositol 3-kinase-driven oncogenic transformation. PMID- 14530395 TI - The dynamics of the spread of selective visual attention. PMID- 14530396 TI - Genetic footprinting of a retroviral Gag gene suggests an important role in virus replication. PMID- 14530397 TI - How an enzyme surmounts the activation energy barrier. PMID- 14530398 TI - Ecological subsidies alter the structure of marine communities. PMID- 14530400 TI - A complex history of rearrangement in an orthologous region of the maize, sorghum, and rice genomes. AB - The sequences of large insert clones containing genomic DNA that is orthologous to the maize adh1 region were obtained for sorghum, rice, and the adh1 homoeologous region of maize, a remnant of the tetraploid history of the Zea lineage. By using all four genomes, it was possible to describe the nature, timing, and lineages of most of the genic rearrangements that have differentiated this chromosome segment over the last 60 million years. The rice genome has been the most stable, sharing 11 orthologous genes with sorghum and exhibiting only one tandem duplication of a gene in this region. The lineage that gave rise to sorghum and maize acquired a two-gene insertion (containing the adh locus), whereas sorghum received two additional gene insertions after its divergence from a common ancestor with maize. The two homoeologous regions of maize have been particularly unstable, with complete or partial deletion of three genes from one segment and four genes from the other segment. As a result, the region now contains only one duplicated locus compared with the eight original loci that were present in each diploid progenitor. Deletion of these maize genes did not remove both copies of any locus. This study suggests that grass genomes are generally unstable in local genome organization and gene content, but that some lineages are much more unstable than others. Maize, probably because of its polyploid origin, has exhibited extensive gene loss so that it is now approaching a diploid state. PMID- 14530399 TI - Dopamine-dependent neurodegeneration in rats induced by viral vector-mediated overexpression of the parkin target protein, CDCrel-1. AB - Mutations in the parkin gene are linked to autosomal-recessive juvenile parkinsonism (AR-JP). Parkin functions as a ubiquitin protein ligase in the degradation of several proteins, including the neuron-specific septin CDCrel-1. AR-JP-associated parkin mutations inhibit ubiquitination and degradation of CDCrel-1 and other parkin target proteins. Here we show that recombinant adeno associated virus-mediated CDCrel-1 gene transfer to the substantia nigra of rats results in a rapid onset (6-10 days) of nigral and striatal CDCrel-1 expression that is followed by a progressive loss of nigral dopaminergic neurons and a decline of the striatal dopamine levels. In contrast, neurons of the globus pallidus are spared from CDCrel-1 toxicity. Furthermore, CDCrel-1 inhibits the release of dopamine from stably-transfected PC12 cells, and pharmacological inhibition of tyrosine hydroxylase and dopamine synthesis in rats prevents CDCrel 1-induced nigral neurodegeneration. These results show that CDCrel-1 overexpression exerts dopamine-dependent neurotoxicity and suggest that inhibition of dopamine secretion by CDCrel-1 may contribute to the development of AR-JP. PMID- 14530401 TI - Kinks, rings, and rackets in filamentous structures. AB - Carbon nanotubes and biological filaments each spontaneously assemble into kinked helices, rings, and "tennis racket" shapes due to competition between elastic and interfacial effects. We show that the slender geometry is a more important determinant of the morphology than any molecular details. Our mesoscopic continuum theory is capable of quantifying observations of these structures and is suggestive of their occurrence in other filamentous assemblies as well. PMID- 14530402 TI - Systemic and cell type-specific gene expression patterns in scleroderma skin. AB - We used DNA microarrays representing >12,000 human genes to characterize gene expression patterns in skin biopsies from individuals with a diagnosis of systemic sclerosis with diffuse scleroderma. We found consistent differences in the patterns of gene expression between skin biopsies from individuals with scleroderma and those from normal, unaffected individuals. The biopsies from affected individuals showed nearly indistinguishable patterns of gene expression in clinically affected and clinically unaffected tissue, even though these were clearly distinguishable from the patterns found in similar tissue from unaffected individuals. Genes characteristically expressed in endothelial cells, B lymphocytes, and fibroblasts showed differential expression between scleroderma and normal biopsies. Analysis of lymphocyte populations in scleroderma skin biopsies by immunohistochemistry suggest the B lymphocyte signature observed on our arrays is from CD20+ B cells. These results provide evidence that scleroderma has systemic manifestations that affect multiple cell types and suggests genes that could be used as potential markers for the disease. PMID- 14530403 TI - Extensive regional atmospheric hydrocarbon pollution in the southwestern United States. AB - Light alkane hydrocarbons are present in major quantities in the near-surface atmosphere of Texas, Oklahoma, and Kansas during both autumn and spring seasons. In spring 2002, maximum mixing ratios of ethane [34 parts per 109 by volume (ppbv)], propane (20 ppbv), and n-butane (13 ppbv) were observed in north-central Texas. The elevated alkane mixing ratios are attributed to emissions from the oil and natural gas industry. Measured alkyl nitrate mixing ratios were comparable to urban smog values, indicating active photochemistry in the presence of nitrogen oxides, and therefore with abundant formation of tropospheric ozone. We estimate that 4-6 teragrams of methane are released annually within the region and represents a significant fraction of the estimated total U.S. emissions. This result suggests that total U.S. natural gas emissions may have been underestimated. Annual ethane emissions from the study region are estimated to be 0.3-0.5 teragrams. PMID- 14530404 TI - How fast is protein hydrophobic collapse? AB - One of the most recurring questions in protein folding refers to the interplay between formation of secondary structure and hydrophobic collapse. In contrast with secondary structure, it is hard to isolate hydrophobic collapse from other folding events. We have directly measured the dynamics of protein hydrophobic collapse in the absence of competing processes. Collapse was triggered with laser induced temperature jumps in the acid-denatured form of a simple protein and monitored by fluorescence resonance energy transfer between probes placed at the protein ends. The relaxation time for hydrophobic collapse is only approximately equal to 60 ns at 305 K, even faster than secondary structure formation. At higher temperatures, as the protein becomes increasingly compact by a stronger hydrophobic force, we observe a slowdown of the dynamics of collapse. This dynamic hydrophobic effect is a high-temperature analogue of the dynamic glass transition predicted by theory. Our results indicate that in physiological conditions many proteins will initiate folding by collapsing to an unstructured globule. Local motions will presumably drive the following search for native structure in the collapsed globule. PMID- 14530405 TI - Activation of NF-kappaB in cells productively infected with HSV-1 depends on activated protein kinase R and plays no apparent role in blocking apoptosis. AB - Microarray data reported elsewhere indicated that herpes simplex virus 1 induces the up-regulation of nuclear factor kappaB (NF-kappaB)-regulated genes, including that of its inhibitor, IkappaBalpha, consistent with the reports that wild-type virus induces the activation of NF-kappaB. In this report we show that activation of NF-kappaB in infected cells is linked to the activation of protein kinase R (PKR). Specifically: (i) PKR is activated in infected cells although the effects of the activated enzyme on protein synthesis are negated by the viral gene gamma134.5, which encodes a protein phosphatase 1alpha accessory factor that enables the dephosphorylation of the alpha subunit of eukaryotic translation initiation factor 2. NF-kappaB is activated in wild-type murine embryonic fibroblasts but not in related PKR-null cells. (ii) In cells infected with a replication-competent Deltagamma134.5 mutant (R5104), but carrying a US11 gene expressed early in infection, eukaryotic translation initiation factor 2alpha is not phosphorylated, and in in vitro assays, PKR bound to the US11 protein is not phosphorylated on subsequent addition of double-stranded RNA. Here we report that this mutant does not activate PKR, has no effect on the accumulation of IkappaBalpha, and does not cause the translocation of NF-kappaB in infected cells. (iii) One hypothesis advanced for the activation of NF-kappaB is that it blocks apoptosis induced by viral gene products. The replication-competent R5104 mutant does not induce the programmed cell's death. We conclude that in herpes simplex virus 1-infected cells, activation of NF-kappaB depends on activation of PKR and that NF-kappaB is not required to block apoptosis in productively infected cells. PMID- 14530406 TI - Topological analysis of a plant vacuolar Na+/H+ antiporter reveals a luminal C terminus that regulates antiporter cation selectivity. AB - We conducted an analysis of the topology of AtNHX1, an Arabidopsis thaliana vacuolar Na+/H+ antiporter. Several hydrophilic regions of the antiporter were tagged with a hemagglutinin epitope, and protease protection assays were conducted to determine the membrane topology of the antiporter by using yeast as a heterologous expression system. The overall structure of AtNHX1 is distinct from the human Na+/H+ antiporter NHE1 or any known Na+/H+ antiporter. It is comprised of nine transmembrane domains and a hydrophilic C-terminal domain. Three hydrophobic regions do not appear to span the tonoplast membrane, yet appear to be membrane associated. Our results also indicate that, whereas the N terminus of AtNHX1 is facing the cytosol, almost the entire C-terminal hydrophilic region resides in the vacuolar lumen. Deletion of the hydrophilic C terminus resulted in a dramatic increase in the relative rate of Na+/H+ transport. The ratio of Na+/K+ transport was twice that of the unmodified AtNHX1. This altered ratio resulted from a relatively small decrease in K+/H+ transport with a large increase in Na+/H+ transport. The vacuolar localization of the C terminus of the AtNHX1, taken together with the regulation of the antiporter selectivity by its C terminus, demonstrates the existence of luminal vacuolar regulatory mechanisms of the antiporter activity. PMID- 14530407 TI - Multiple shifts in the representation of a motor sequence during the acquisition of skilled performance. AB - When do learning-related changes in performance occur? Here we show that the knowledge of a sequence of movements evolves through several distinctive phases that depend on two critical factors: the amount of practice as well as the passage of time. Our results show the following. (i) Within a given session, large performance gains constituted a signature for motor novelty. Such gains occurred only for newly introduced conditions irrespective of the absolute level of performance. (ii) A single training session resulted in both immediate but also time-dependent, latent learning hours after the termination of practice. Time in sleep determined the time of expression of these delayed gains. Moreover, the delayed gains were sequence-specific, indicating a qualitative change in the representation of the task within 24 h posttraining. (iii) Prolonged training resulted in additional between-session gains that, unlike the effects of a single training session, were confined to the trained hand. Thus, the effects of multisession training were qualitatively different than the immediate and time dependent effects of a single session. Altogether, our results indicate multiple time-dependent shifts in the representation of motor experience during the acquisition of skilled performance. PMID- 14530408 TI - Facile synthesis of site-specifically acetylated and methylated histone proteins: reagents for evaluation of the histone code hypothesis. AB - The functional capacity of genetically encoded histone proteins can be powerfully expanded by posttranslational modification. A growing body of biochemical and genetic evidence clearly links the unique combinatorial patterning of side chain acetylation, methylation, and phosphorylation mainly within the highly conserved N termini of histones H2A, H2B, H3, and H4 with the regulation of gene expression and chromatin assembly and remodeling, in effect constituting a "histone code" for epigenetic signaling. Deconvoluting this code has proved challenging given the inherent posttranslational heterogeneity of histone proteins isolated from biological sources. Here we describe the application of native chemical ligation to the preparation of full-length histone proteins containing site-specific acetylation and methylation modifications. Peptide thioesters corresponding to histone N termini were prepared by solid phase peptide synthesis using an acid labile Boc/HF assembly strategy, then subsequently ligated to recombinantly produced histone C-terminal globular domains containing an engineered N-terminal cysteine residue. The ligation site is then rendered traceless by hydrogenolytic desulfurization, generating a native histone protein sequence. Synthetic histones generated by this method are fully functional, as evidenced by their self assembly into a higher order H3/H4 heterotetramer, their deposition into nucleosomes by human ISWI-containing (Imitation of Switch) factor RSF (Remodeling and Spacing Factor), and by enzymatic modification by human Sirt1 deacetylase and G9a methyltransferase. Site-specifically modified histone proteins generated by this method will prove invaluable as novel reagents for the evaluation of the histone code hypothesis and analysis of epigenetic signaling mechanisms. PMID- 14530409 TI - Repeated neonatal handling with maternal separation permanently alters hippocampal GABAA receptors and behavioral stress responses. AB - Increasing evidence suggests that postnatal events, such as handling or maternal separation, can produce long-term changes in brain function. These are often expressed as changes in the profile of endocrine or behavioral responses to stress. Changes in gamma-aminobutyric acid type A receptors (GABARs), which mediate the majority of fast synaptic inhibition in adult brain, have been proposed as one potential mediator of these behavioral effects. In the current article, we use a combination of single-cell electrophysiology and antisense mRNA amplification to demonstrate permanent molecular and functional differences in GABARs within hippocampal dentate granule neurons after as few as two episodes of neonatal handling with brief maternal separation. Adult animals that as pups experienced handling with maternal separation maintained a more immature GABAR phenotype and exhibited increased activity in response to swim stress. These findings demonstrate the exquisite sensitivity of the developing GABAergic system to even subtle environmental manipulations and provide an unique molecular mechanism by which postnatal handling with maternal separation may alter stress related behavior. PMID- 14530410 TI - GABA uptake into astrocytes is not associated with significant metabolic cost: implications for brain imaging of inhibitory transmission. AB - Synaptically released glutamate has been identified as a signal coupling excitatory neuronal activity to increased glucose utilization. The proposed mechanism of this coupling involves glutamate uptake into astrocytes resulting in increased intracellular Na+ (Nai+) and activation of the Na+/K+-ATPase. Increased metabolic demand linked to disruption of Nai+ homeostasis activates glucose uptake and glycolysis in astrocytes. Here, we have examined whether a similar neurometabolic coupling could operate for the inhibitory neurotransmitter gamma aminobutyric acid (GABA), also taken up by Na+-dependent transporters into astrocytes. Thus, we have compared the Nai+ response to GABA and glutamate in mouse astrocytes by microspectrofluorimetry. The Nai+ response to GABA consisted of a rapid rise of 4-6 mM followed by a plateau that did not, however, significantly activate the pump. Indeed, the GABA transporter-evoked Na+ influxes are transient in nature, almost totally shutting off within approximately 30 sec of GABA application. The metabolic consequences of the GABA-induced Nai+ response were evaluated by monitoring cellular ATP changes indirectly in single cells and measuring 2-deoxyglucose uptake in astrocyte populations. Both approaches showed that, whereas glutamate induced a robust metabolic response in astrocytes (decreased ATP levels and glucose uptake stimulation), GABA did not cause any measurable metabolic response, consistent with the Nai+ measurements. Results indicate that GABA does not couple inhibitory neuronal activity with glucose utilization, as does glutamate for excitatory neurotransmission, and suggest that GABA-mediated synaptic transmission does not contribute directly to brain imaging signals based on deoxyglucose. PMID- 14530411 TI - Cardiac progenitor cells from adult myocardium: homing, differentiation, and fusion after infarction. AB - Potential repair by cell grafting or mobilizing endogenous cells holds particular attraction in heart disease, where the meager capacity for cardiomyocyte proliferation likely contributes to the irreversibility of heart failure. Whether cardiac progenitors exist in adult myocardium itself is unanswered, as is the question whether undifferentiated cardiac precursor cells merely fuse with preexisting myocytes. Here we report the existence of adult heart-derived cardiac progenitor cells expressing stem cell antigen-1. Initially, the cells express neither cardiac structural genes nor Nkx2.5 but differentiate in vitro in response to 5'-azacytidine, in part depending on Bmpr1a, a receptor for bone morphogenetic proteins. Given intravenously after ischemia/reperfusion, cardiac stem cell antigen 1 cells home to injured myocardium. By using a Cre/Lox donor/recipient pair (alphaMHC-Cre/R26R), differentiation was shown to occur roughly equally, with and without fusion to host cells. PMID- 14530415 TI - The William Houston Gold Medal Prize in the M.Orth. Exam of the Royal College of Surgeons of Edinburgh held in Edinburgh, 2000. PMID- 14530412 TI - Membrane association of myotubularin-related protein 2 is mediated by a pleckstrin homology-GRAM domain and a coiled-coil dimerization module. AB - Mutations in the myotubularin (MTM)-related protein 2 (MTMR2) gene are responsible for the severe autosomal recessive neuropathy Charcot-Marie-Tooth disease type 4B1. MTMR2 belongs to the MTM family of dual-specific phosphatases that use phosphatidylinositol (PI) 3,5-bisphosphate [PI(3,5)P2] and PI 3 phosphate [PI(3)P] as their substrate. Because these substrates are localized in the membrane bilayer, membrane targeting of Mtmr2 is an important regulatory mechanism. In hypoosmotically stressed COS cells with increased levels of PI(3,5)P2, Mtmr2 is bound to the membrane of vacuoles formed under these conditions. Using several mutant forms of Mtmr2, we identified two domains that are necessary for membrane association: (i) A pleckstrin homology-GRAM domain; and (ii) a coiled-coil module. Protein-lipid overlay assays show that the pleckstrin homology-GRAM domain binds to PI(3,5)P2 and PI(5)P, a substrate and a product of the Mtmr2 enzyme, respectively. We also demonstrate that Mtmr2 forms a dimer and that the C-terminal coiled-coil is responsible for homodimerization, in addition to membrane association. Our data indicate that phosphoinositide-protein interactions, as well as protein-protein interactions, are necessary for the correct regulation of MTMR2. PMID- 14530413 TI - Enzyme screening with synthetic multifunctional pores: focus on biopolymers. AB - This report demonstrates that a single set of identical synthetic multifunctional pores can detect the activity of many different enzymes. Enzymes catalyzing either synthesis or degradation of DNA (exonuclease III or polymerase I), RNA (RNase A), polysaccharides (heparinase I, hyaluronidase, and galactosyltransferase), and proteins (papain, ficin, elastase, subtilisin, and pronase) are selected to exemplify this key characteristic of synthetic multifunctional pore sensors. Because anionic, cationic, and neutral substrates can gain access to the interior of complementarily functionalized pores, such pores can be the basis for very user-friendly screening of a broad range of enzymes. PMID- 14530416 TI - Class III Twin Blocks: a case series. AB - OBJECTIVE: To evaluate the use of Class III Twin Blocks for the early treatment of Class III malocclusion. DESIGN: Retrospective analysis. SUBJECTS AND METHOD: 14 subjects were consecutively treated with a modified version of the Class III Twin Block appliance. Lateral cephalometric radiographs taken at the start and end of treatment were assessed digitally. RESULTS: The mean age of subjects was 10 years and mean treatment time was 6.6 months. The cephalometric analysis of changes during treatment shows proclination of the upper incisors [mean 5.1 degrees, SD 5.1], retroclination of the lower incisors [mean -4.5 degrees, SD 3.9], reduction in angle SNB [mean -1.3 degrees, SD 1.8] and an increase in the maxillary/ mandibular planes angle (mean 2.1 degrees, SD 2.2]. Average cephalometric digitizations and photographs of a clinical case are shown. CONCLUSIONS: Class III Twin Blocks can be used successfully for early treatment of Class III malocclusion. A randomly allocated prospective study is required to fully evaluate the efficacy of this appliance. PMID- 14530417 TI - Pseudo-Class III malocclusion treatment with Balters' Bionator. AB - The aim of this article is to show the use of the Balters' Bionator in pseudo Class III treatment. The importance of differentiating between true Class III and pseudo-Class III is emphasized. The therapeutic results of a Balters' Bionator appliance are presented in three case reports of subjects in the mixed dentition. In this stage of development it is possible to correct an isolated problem. The use of the Bionator III in this kind of malocclusion enabled the correction of a dental malocclusion in a few months and therapeutic stability of a mesially positioned mandible encouraging favourable skeletal growth. PMID- 14530419 TI - Assessment of the accuracy of a three-dimensional imaging system for archiving dental study models. AB - OBJECTIVE: The use of stone and plaster study models is an integral part of any dental practice and is required for research. Storage of study models is problematic in terms of space and cost. Ayoub et al.(1) introduced a new technique based on the recent advances in stereophotogrammetry for archiving dental study models in a digital format. However, assessment of the accuracy of the generated three-dimensional (3D) models has not been carried out yet. It was the aim of this study to evaluate the accuracy of this technique. DESIGN: A comparative assessment between direct measurements of dental study models and measurements of computer generated 3D images of the same study models was performed. MATERIALS AND METHODS: Twenty-two dental study models stored at Glasgow Dental Hospital and School for the purposes of research were used in the study. The models were captured in three dimensions using a photostereometric technique and stored in digital format. MAIN OUTCOME MEASURES: Measurements were conducted directly on dental study models and on the computer generated 3D images using Euclidean Distance Matrix Analysis.(2) The difference between the two sets of measurements was statistically analysed using a two-sample t-test. RESULTS: The average difference between measurements of dental casts and 3D images was 0.27 mm. This difference was within the range of operator errors (0.10-0.48 mm) and was not statistically significant (P < 0.05). CONCLUSION: This study shows that it is possible to use 3D imaging to store dental study models for treatment monitoring and research with a satisfactory degree of accuracy. PMID- 14530420 TI - An ex vivo assessment of a bonding technique using a self-etching primer. AB - OBJECTIVE: This study assessed a new self-etch/priming system for use in orthodontic bonding. SETTING: An ex vivo study. METHOD: Three groups of 20 extracted premolar teeth were bonded with metal orthodontic brackets. Group 1 was bonded with Transbond using the conventional technique (control). Group 2 was bonded using the new Transbond-Plus combined etch/primer system to wet enamel and Group 3 to dry enamel. The teeth were debonded using an Instron Universal Testing Machine. The mean debond force was calculated for each group and compared statistically. The teeth were examined under the stereomicroscope to assess the site of debond and adhesive remnant index. RESULTS: Group 2 (etch/primer on wet enamel) had the lowest mean debond value at 5.2 MPa. ANOVA and Tukey tests confirmed that the bond strength results of Group 2 were significantly lower than Groups 1 (P < 0.01) and 3 (P < 0.05). The enamel/resin interface was the commonest site of bond failure for both etch/primer groups (Groups 2 and 3). They had less retained resin and significantly (P < 0.001) lower ARI scores compared with Group 1 (control). CONCLUSIONS: The results of this ex vivo study suggest that the self-etch primer should achieve adequate bond strengths when applied to dry enamel surfaces. In addition there should be less retained resin requiring removal at debond. PMID- 14530421 TI - A clinical investigation of force delivery systems for orthodontic space closure. AB - OBJECTIVE: To investigate the force retention, and rates of space closure achieved by elastomeric chain and nickel titanium coil springs. DESIGN: Randomized clinical trial. SETTING: Eastman Dental Hospital, London and Queen Mary's University Hospital, Roehampton, 1998-2000. SUBJECTS, MATERIALS AND METHODS: Twenty-two orthodontic patients, wearing the pre-adjusted edgewise appliance undergoing space closure in opposing quadrants, using sliding mechanics on 0.019 x 0.025-inch posted stainless steel archwires. Medium-spaced elastomeric chain [Durachain, OrthoCare (UK) Ltd., Bradford, UK] and 9-mm nickel titanium coil springs [OrthoCare (UK) Ltd.] were placed in opposing quadrants for 15 patients. Elastomeric chain only was used in a further seven patients. The initial forces on placement and residual forces at the subsequent visit were measured with a dial push-pull gauge [Orthocare (UK) Ltd]. Study models of eight patients were taken before and after space closure, from which measurements were made to establish mean space closure. MAIN OUTCOME MEASURES: The forces were measured in grammes and space closure in millimetres. RESULTS: Fifty-nine per cent (31/53) of the elastomeric sample maintained at least 50 per cent of the initial force over a time period of 1-15 weeks. No sample lost all its force, and the mean loss was 47 per cent (range: 0-76 per cent). Nickel titanium coil springs lost force rapidly over 6 weeks, following that force levels plateaued. Forty-six per cent (12/26) maintained at least 50 per cent of their initial force over a time period of 1-22 weeks, and mean force loss was 48 per cent (range: 12 68 per cent). The rate of mean weekly space closure for elastomeric chain was 0.21 mm and for nickel titanium coil springs 0.26 mm. There was no relationship between the initial force applied and rate of space closure. None of the sample failed during the study period giving a 100 per cent response rate. CONCLUSIONS: In clinical use, the force retention of elastomeric chain was better than previously concluded. High initial forces resulted in high force decay. Nickel titanium coil springs and elastomeric chain closed spaces at a similar rate. PMID- 14530422 TI - Orthodontic treatment outcome: the relationship between anterior dental relations and anterior inter-arch tooth size discrepancy. AB - OBJECTIVE: This study investigated the pre- (T1) and post-treatment (T2) relationship between anterior (canine to canine inclusive) inter-arch tooth size ratio and various dental and skeletal variables. DESIGN: Retrospective longitudinal clinical study. SETTING: Swedish Health Board Clinic 2000. SUBJECTS: Random selection of T1 and T2 orthodontic records of 137 Swedish patients (56 male and 81 female). The sample included non-extraction (77), and four premolar extraction (60) cases across a range of dental and skeletal malrelationships. MAIN OUTCOME MEASURES: Dental cast and lateral cephalogram measurements were recorded. Exploratory modelling investigated whether a significant relationship existed between the anterior inter-arch tooth size ratio and these measurements. RESULTS: Data was normally distributed with no statistically significant differences between males and females (P = 0.88) and extraction and non extraction (P = 0.52) treatment modalities with respect to the anterior ratio. T1 bivariate regression analysis failed to show a relationship (p < 0.05) between variables. T2 bivariate analysis showed a statistically significant relation between three variables and anterior tooth size ratio. Multiple regression analysis led to a final model where maxillary inter-canine width (P = 0.002) and upper arch crowding (0.001) were statistically significantly related to the anterior inter-arch ratio. The coefficient of determination was however uniformly low (R(2) < 0.2) for all variables. CONCLUSION: The anterior inter-arch tooth size ratio was not associated with any common pre- or post-treatment variables in the population studied, therefore measurement of an anterior tooth size ratio pre treatment was not clinically beneficial for determining anterior dental relations post-treatment. PMID- 14530423 TI - How to...write and analyse a questionnaire. AB - Questionnaires are used in a wide range of settings to gather information about the opinions and behaviour of individuals. As with any other branch of science, the validity and reliability of the measurement tool, i.e. the questionnaire, needs to be rigorously tested to ensure that the data collected is meaningful. The design and method of administration of a questionnaire will also influence the response rate that is achieved and the quality of data that is collected. This paper discusses the issues that should be considered when designing and undertaking a questionnaire study. PMID- 14530427 TI - Self-ligating brackets: where are we now? AB - The current situation regarding self-ligating brackets is reviewed. Recent developments, clinical advantages, and remaining imperfections are described. The question of active versus passive ligation is scrutinized. The evidence regarding treatment efficiency is reviewed. Suggested clinical tips and changes of treatment mechanics are summarized and illustrated. Self-ligating brackets have reached a stage of design and production control, where the advantages are significantly greater than the remaining imperfections. PMID- 14530428 TI - Does articulating study casts make a difference to treatment planning. PMID- 14530429 TI - Differential transcription factor occupancy but evolutionarily conserved chromatin features at the human and mouse M-CSF (CSF-1) receptor loci. AB - The c-FMS gene encodes the macrophage colony-stimulating factor receptor (M-CSFR or CSF1-R), which is a tyrosine kinase growth factor receptor essential for macrophage development. We have previously characterized the chromatin features of the mouse gene; however, very little is known about chromatin structure and function of the human c-FMS locus. Here we present a side-by-side comparison of the chromatin structure, histone modification, transcription factor occupancy and cofactor recruitment of the human and the mouse c-FMS loci. We show that, similar to the mouse gene, the human c-FMS gene possesses a promoter and an intronic enhancer element (c-fms intronic regulatory element or FIRE). Both elements are evolutionarily conserved and specifically active in macrophages. However, we demonstrate by in vivo footprinting that both murine and human c-FMS cis regulatory elements are recognised by an overlapping, but non-identical, set of transcription factors. Despite these differences, chromatin immunoprecipitation experiments show highly similar patterns of histone H3 modification and a similar distribution of chromatin modifying and remodelling activities at individual cis regulatory elements and across the c-FMS locus. Our experiments support the hypothesis that the same regulatory principles operate at both genes via conserved cores of transcription factor binding sites. PMID- 14530430 TI - Use of locked nucleic acid oligonucleotides to add functionality to plasmid DNA. AB - The available reagents for the attachment of functional moieties to plasmid DNA are limiting. Most reagents bind plasmid DNA in a non-sequence- specific manner, with undefined stoichiometry, and affect DNA charge and delivery properties or involve chemical modifications that abolish gene expression. The design and ability of oligonucleotides (ODNs) containing locked nucleic acids (LNAs) to bind supercoiled, double-stranded plasmid DNA in a sequence-specific manner are described for the first time. The main mechanism for LNA ODNs binding plasmid DNA is demonstrated to be by strand displacement. LNA ODNs are more stably bound to plasmid DNA than similar peptide nucleic acid (PNA) 'clamps' for procedures such as particle-mediated DNA delivery (gene gun). It is shown that LNA ODNs remain associated with plasmid DNA after cationic lipid-mediated transfection into mammalian cells. LNA ODNs can bind to DNA in a sequence-specific manner so that binding does not interfere with plasmid conformation or gene expression. Attachment of CpG-based immune adjuvants to plasmid by 'hybrid' phosphorothioate LNA ODNs induces tumour necrosis factor-alpha production in the macrophage cell line RAW264.7. This observation exemplifies an important new, controllable methodology for adding functionality to plasmids for gene delivery and DNA vaccination. PMID- 14530431 TI - Synergistic repression of anaerobic genes by Mot3 and Rox1 in Saccharomyces cerevisiae. AB - Two groups of anaerobic genes (genes induced in anaerobic cells and repressed in aerobic cells) are negatively regulated by heme, a metabolite present only in aerobic cells. Members of both groups, the hypoxic genes and the DAN/TIR/ERG genes, are jointly repressed under aerobic conditions by two factors. One is Rox1, an HMG protein, and the second, originally designated Rox7, is shown here to be Mot3, a global C2H2 zinc finger regulator. Repression of anaerobic genes results from co-induction of Mot3 and Rox1 in aerobic cells. Repressor synthesis is triggered by heme, which de-represses a mechanism controlling expression of both MOT3 and ROX1 in anaerobic cells; it includes Hap1, Tup1, Ssn6 and a fourth unidentified factor. The constitutive expression of various anaerobic genes in aerobic rox1Delta or mot3Delta cells directly implies that neither factor can repress by itself at endogenous levels and that stringent aerobic repression results from the concerted action of both. Mot3 and Rox1 are not essential components of a single complex, since each can repress independently in the absence of the other, when artificially induced at high levels. Moreover, the two repression mechanisms appear to be distinct: as shown here repression of ANB1 by Rox1 alone requires Tup1-Ssn6, whereas repression by Mot3 does not. Though artificially high levels of either factor can repress well, the absolute efficiency observed in normal cells when both are present-at much lower levels demonstrates a novel inhibitory synergy. Evidently, expression levels for the two mutually dependent repressors are calibrated to permit a range of variation in basal aerobic expression at different promoters with differing operator site combinations. PMID- 14530432 TI - Genetic and biochemical interactions between SCP160 and EAP1 in yeast. AB - Scp160p is a multiple KH-domain RNA-binding protein in yeast known to associate with polyribosomes as an mRNP component, although its biological role remains unclear. As a genetic approach to examine Scp160p function, we applied an ethyl methanesulfonate (EMS) screen for loci synthetically lethal with scp160 loss, and identified a single candidate gene, EAP1, whose protein product functions in translation as an eIF4E-binding protein, with additional uncharacterized spindle pole body functions. To reconfirm scp160/eap1 synthetic lethality, we constructed a strain null for both genes, supported by an SCP160 maintenance plasmid. We used this strain to establish a quantitative assay for both Scp160p and Eap1p functions in vivo, and applied this assay to demonstrate that Y109A EAP1, a previously described allele of EAP1 that cannot bind eIF4E, is markedly impaired with regard to its SCP160-related activity. In addition, we explored the possibility of physical interaction between Eap1p and Scp160p, and discovered that Eap1p associates with Scp160p-containing complexes in an RNA-dependent manner. Finally, we probed the impact of EAP1 loss on Scp160p, and vice versa, and found that loss of each gene resulted in a significant change in either the complex associations or subcellular distribution of the other protein. These results clearly support the hypothesis that Scp160p plays a role in translation, demonstrate that the interaction of SCP160 and EAP1 is biologically significant, and provide important tools for future studies of the in vivo functions of both genes. PMID- 14530433 TI - Modification of the ionizing radiation response in living cells by an scFv against the DNA-dependent protein kinase. AB - The non-homologous end joining pathway uses pre-existing proteins to repair DNA double-strand breaks induced by ionizing radiation. Here we describe manipulation of this pathway in living cells using a newly developed tool. We generated a single chain antibody variable fragment (scFv) that binds to the DNA-dependent protein kinase catalytic subunit (DNA-PKcs), a key enzyme in the pathway. In contrast to existing pharmacologic inhibitors, the scFv binds a newly defined regulatory site outside the kinase catalytic domain. Although the scFv inhibits kinase activity only modestly, it completely blocks DNA end joining in a cell free system. Microinjection of the scFv sensitizes human cells to radiation, as measured by a reduction in efficiency of colony formation and induction of apoptosis at an otherwise sublethal dose of 1.5 Gy. The scFv blocks non homologous end joining in situ at a step subsequent to histone gamma-H2AX focus formation but preceding gamma-H2AX dephosphorylation. Blockage occurs in cells exposed to as little as 0.1 Gy, indicating that DNA-PKcs is essential for double strand break repair even at low radiation doses. The ability to modify the radiation response in situ in living cells provides a link between biochemical, genetic and cytologic approaches to the study of double-strand break repair intermediates. PMID- 14530434 TI - NMR structure of an alpha-L-LNA:RNA hybrid: structural implications for RNase H recognition. AB - Alpha-L-LNA (alpha-L-ribo configured locked nucleic acid) is a nucleotide analogue that raises the thermostability of nucleic acid duplexes by up to approximately 4 degrees C per inclusion. We have determined the NMR structure of a nonamer alpha-L-LNA:RNA hybrid with three alpha-L-LNA modifications. The geometry of this hybrid is intermediate between A- and B-type, all nucleobases partake in Watson-Crick base pairing and base stacking, and the global structure is very similar to that of the corresponding unmodified hybrid. The sugar phosphate backbone is rearranged in the vicinity of the modified nucleotides. As a consequence, the phosphate groups following the modified nucleotides are rotated into the minor groove. It is interesting that the alpha-L-LNA:RNA hybrid, which has an elevation in melting temperature of 17 degrees C relative to the corresponding DNA:RNA hybrid, retains the global structure of this hybrid. To our knowledge, this is the first example of such a substantial increase in melting temperature of a nucleic acid analogue that does not act as an N-type (RNA) mimic. alpha-L-LNA:RNA hybrids are recognised by RNase H with subsequent cleavage of the RNA strand, albeit with slow rates. We attempt to rationalise this impaired enzyme activity from the rearrangement of the sugar-phosphate backbone of the alpha-L-LNA:RNA hybrid. PMID- 14530435 TI - The positive and negative regulation of Tn10 transposition by IHF is mediated by structurally asymmetric transposon arms. AB - The Tn10 transpososome has symmetrical components on either side: there are two transposon ends each of which has binding sites for a monomer of transposase and an IHF heterodimer. The DNA bending activity of IHF stimulates assembly of an intermediate with tightly folded transposon ends in which transposase has additional 'subterminal' DNA contacts, located distal to the IHF site. These subterminal contacts are required to activate later steps in the reaction. Quantitative hydroxyl radical footprinting and gel retardation unfolding experiments show that the transpososome is fundamentally asymmetric, despite having identical components on either side. Major differences between the transposon ends define alpha and beta sides of the complex. IHF can dissociate from the transposon arm on the beta side of the complex in the absence of metal ion. However, IHF is locked onto the alpha side of the complex, probably by the subterminal transposase contacts, until released by a metal ion-dependent conformational change. Later in the reaction, IHF inhibits target interactions. Using a very short transposon arm, target interactions are demonstrated at a saturating IHF concentration. This suggests that inhibition of target interactions is due to steric hindrance of the target binding site by a single IHF-folded transposon arm. PMID- 14530436 TI - An organism-specific method to rank predicted coding regions in Trypanosoma brucei. AB - Genome annotation in differently evolved organisms presents challenges because the lack of sequence-based homology limits the ability to determine the function of putative coding regions. To provide an alternative to annotation by sequence homology, we developed a method that takes advantage of unusual trypanosomatid biology and skews in nucleotide composition between coding regions and upstream regions to rank putative open reading frames based on the likelihood of coding. The method is 93% accurate when tested on known genes. We have applied our method to the full complement of open reading frames on Chromosome I of Trypanosoma brucei, and we can predict with high confidence that 226 putative coding regions are likely to be functional. Methods such as the one described here for discriminating true coding regions are critical for genome annotation when other sources of evidence for function are limited. PMID- 14530437 TI - Modulation of gene expression in Leishmania drug resistant mutants as determined by targeted DNA microarrays. AB - In the protozoan parasite Leishmania, drug resistance can be a complex phenomenon. Several metabolic pathways and membrane transporters are implicated in the resistance phenotype. To monitor the expression of these genes, we generated custom DNA microarrays with PCR fragments corresponding to 44 genes involved with drug resistance. Transcript profiling of arsenite and antimony resistant mutants with these arrays pinpointed a number of genes overexpressed in mutants, including the ABC transporter PGPA, the glutathione biosynthesis genes gamma-glutamylcysteine synthetase (GSH1) and the glutathione synthetase (GSH2). Competitive hybridisations with total RNA derived from sensitive and methotrexate resistant cells revealed the overexpression of genes coding for dihydrofolate reductase (DHFR-TS), pteridine reductase (PTR1) and S-adenosylmethionine synthase (MAT2) and a down regulation of one gene of the folate transporter (FT) family. By labelling the DNA of sensitive and resistant parasites we could also detect several gene amplification events using DNA microarrays including the amplification of the S-adenosyl homocysteine hydrolase gene (SAHH). Alteration in gene expression detected by microarrays was validated by northern blot analysis, while Southern blots indicated that most genes overexpressed were also amplified, although other mechanisms were also present. The microarrays were useful in the study of resistant parasites to pinpoint several genes linked to drug resistance. PMID- 14530438 TI - Effects of genomic context and chromatin structure on transcription-coupled and global genomic repair in mammalian cells. AB - It has been long recognized that in mammalian cells, DNA damage is preferentially repaired in the transcribed strand of transcriptionally active genes. However, recently, we found that in Chinese hamster ovary (CHO) cells, UV-induced cyclobutane pyrimidine dimers (CPDs) are preferentially repaired in both the transcribed and the non-transcribed strand of exon 1 of the dihydrofolate reductase (DHFR) gene. We mapped CPD repair at the nucleotide level in the transcriptionally active DHFR gene and the adjacent upstream OST gene, both of which have been translocated to two chromosomal positions that differ from their normal endogeneous positions. This allowed us to study the role of transcription, genomic context and chromatin structure on repair. We found that CPD repair in the transcribed strand is the same for endogenous and translocated DHFR genes, and the order of repair efficiency is exon 1 > exon 2 > exon 5. However, unlike the endogenous DHFR gene, efficient repair of CPDs in the non-transcribed strand of exon 1 is not observed in the translocated DHFR gene. CPDs are efficiently repaired in the transcribed strand in endogenous and translocated OST genes, which indicates that efficient repair in exon 1 of the non-transcribed strand of the endogenous DHFR gene is not due to the extension of transcription-coupled repair of the OST gene. Using micrococcal nuclease digestion, we probed the chromatin structure in the DHFR gene and found that chromatin structure in the exon 1 region of endogenous DHFR is much more open than at translocated loci. These results suggest that while transcription-coupled repair is transcription dependent, global genomic repair is greatly affected by chromatin structure. PMID- 14530439 TI - The complete nucleotide sequence and RNA editing content of the mitochondrial genome of rapeseed (Brassica napus L.): comparative analysis of the mitochondrial genomes of rapeseed and Arabidopsis thaliana. AB - The entire mitochondrial genome of rapeseed (Brassica napus L.) was sequenced and compared with that of Arabidopsis thaliana. The 221 853 bp genome contains 34 protein-coding genes, three rRNA genes and 17 tRNA genes. This gene content is almost identical to that of Arabidopsis: However the rps14 gene, which is a pseudo-gene in Arabidopsis, is intact in rapeseed. On the other hand, five tRNA genes are missing in rapeseed compared to Arabidopsis, although the set of mitochondrially encoded tRNA species is identical in the two Cruciferae. RNA editing events were systematically investigated on the basis of the sequence of the rapeseed mitochondrial genome. A total of 427 C to U conversions were identified in ORFs, which is nearly identical to the number in Arabidopsis (441 sites). The gene sequences and intron structures are mostly conserved (more than 99% similarity for protein-coding regions); however, only 358 editing sites (83% of total editings) are shared by rapeseed and Arabidopsis: Non-coding regions are mostly divergent between the two plants. One-third (about 78.7 kb) and two-thirds (about 223.8 kb) of the rapeseed and Arabidopsis mitochondrial genomes, respectively, cannot be aligned with each other and most of these regions do not show any homology to sequences registered in the DNA databases. The results of the comparative analysis between the rapeseed and Arabidopsis mitochondrial genomes suggest that higher plant mitochondria are extremely conservative with respect to coding sequences and somewhat conservative with respect to RNA editing, but that non-coding parts of plant mitochondrial DNA are extraordinarily dynamic with respect to structural changes, sequence acquisition and/or sequence loss. PMID- 14530440 TI - Hexameric RSF1010 helicase RepA: the structural and functional importance of single amino acid residues. AB - In the known monoclinic crystals the 3-dimensional structure of the hexameric, replicative helicase RepA encoded by plasmid RSF1010 shows 6-fold rotational symmetry. In contrast, in the cubic crystal form at 2.55 A resolution described here RepA has 3-fold symmetry and consists of a trimer of dimers. To study structure-function relationships, a series of repA deletion mutants and mutations yielding single amino acid exchanges were constructed and the respective gene products were analyzed in vivo and in vitro. Hexamerization of RepA occurs via the N-terminus and is required for NTP hydrolysis. The C-terminus is essential both for the interaction with the replication machinery and for the helicase activity. Functional analyses of RepA variants with single amino acid exchanges confirmed most of the predictions that were based on the published 3-dimensional structure. Of the five motifs conserved in family 4 helicases, all residues conserved in RepA and T7 gp4 helicases participate in DNA unwinding. Residues K42, E76, D77, D139 and H178, proposed to play key roles in catalyzing the hydrolysis of NTPs, are essential for RepA activity. Residue H178 of motif H3 couples nucleotide consumption to DNA strand separation. PMID- 14530441 TI - Structural study of DNA duplex containing an N-(2-deoxy-beta-D-erythro pentofuranosyl) formamide frameshift by NMR and restrained molecular dynamics. AB - The presence of an N-(2-deoxy-beta-D-erythro-pentofuranosyl) formamide (F) residue, a ring fragmentation product of thymine, in a frameshift context in the sequence 5'-d-(AGGACCACG)*d(CGTGGFTCCT) has been studied by 1H and 31P nuclear magnetic resonance (NMR) and molecular dynamics. Two-dimensional NMR studies show that the formamide residue, whether the cis or trans isomer, is rotated out of the helix and that the bases on either side of the formamide residue in the sequence, G14 and T16, are stacked over each other in a way similar to normal B DNA. The cis and trans isomers were observed in the ratio 3:2 in solution. Information extracted from 31P NMR data reveal a modification of the phosphodiester backbone conformation at the extrahelical site, which is also observed during the molecular dynamics simulations. PMID- 14530443 TI - Antizyme frameshifting as a functional probe of eukaryotic translational termination. AB - Translation termination in eukaryotes is mediated by the release factors eRF1 and eRF3, but mechanisms of the interplay between these factors are not fully understood, due partly to the difficulty of measuring termination on eukaryotic mRNAs. Here, we describe an in vitro system for the assay of termination using competition with programmed frameshifting at the recoding signal of mammalian antizyme. The efficiency of antizyme frameshifting in rabbit reticulocyte lysates was reduced by addition of recombinant rabbit eRF1 and eRF3 in a synergistic manner. Addition of suppressor tRNA to this assay system revealed competition with a third event, stop codon readthrough. Using these assays, we demonstrated that an eRF3 mutation at the GTPase domain repressed termination in a dominant negative fashion probably by binding to eRF1. The effect of the release factors and the suppressor tRNA showed that the stop codon at the antizyme frameshift site is relatively inefficient compared to either the natural termination signals at the end of protein coding sequences or the readthrough signal from a plant virus. The system affords a convenient assay for release factor activity and has provided some novel views of the mechanism of antizyme frameshifting. PMID- 14530442 TI - Sox6 regulation of cardiac myocyte development. AB - A mouse mutation (p100H/p100H) has been identified that is associated with cardioskeletal myopathy, heart block, delayed growth and early postnatal death. The gene that is disrupted in this mutation encodes the transcription factor Sox6. P19CL6 cells were used as an in vitro cardiomyocyte differentiation system and revealed that Sox6 is expressed exclusively when the cells are committed to differentiate to beating cardiac myocytes. We used the yeast two-hybrid system to identify the Prtb (Proline-rich transcript of the brain) protein as a Sox6 interactor, and subsequently confirmed the interaction by co-immunoprecipitation. Prtb expression in P19CL6 cells increased with differentiation to beating cardiomyocytes. Using the P19CL6 cells stably transfected with noggin, an antagonist of BMP (Bone Morphogenic Protein), we found that BMP expression is required for Sox6 expression in cardiomyocyte differentiation. Surprisingly, the expression of the alpha1c-subunit gene of the L-type Ca2+ channel decreased in P19CL6 cells as they differentiated to beating cardiac cells. Ectopic expression of Sox6 or Prtb alone in P19CL6 cells caused down-regulation of L-type Ca2+ alpha1c expression, but when Sox6 and Prtb were co-transfected to the cells, L type Ca2+ alpha1c remained at basal levels. A similar relationship of Sox6 and L type Ca2+ alpha1c expression was seen in vivo (comparing wild-type and p(100H)/p(100H) mutant mice). Thus, Sox6 is within the BMP pathway in cardiac differentiation, interacts with Prtb and may play a critical role in the regulation of a cardiac L-type Ca2+ channel. PMID- 14530444 TI - The inflammation-induced down-regulation of plasma Fetuin-A (alpha2HS Glycoprotein) in liver results from the loss of interaction between long C/EBP isoforms at two neighbouring binding sites. AB - Fetuin-A is an hepatic protein whose mRNA transiently falls during the inflammatory acute phase via unknown transcriptional mechanisms. Various FETUA promoter/cat constructs transiently transfected in the Hep3B hepatoma cell line allowed us to identify four NF-1 and C/EBP binding sites (N, C) arranged in a 5' N2-C2-N1-C1-3' order and required for basal promoter activity. Mutant constructs demonstrated that C1 and C2 but not N1 nor N2 are required for the cytokine driven down-regulation of the promoter. A variable spacing between C2 and N1 showed that the alignment of the (C1+N1) and (C2+N2) areas is critical for the promoter activity in quiescent but not cytokine-stimulated cells. Co-transfection of a plasmid only producing either a long or short C/EBPbeta isoform prevented FETUA regulation by cytokines. Electromobility shift assays with liver nuclear extracts showed that during the acute phase the complexes formed over N1 and N2 are not modified whereas short C/EBPalpha and -beta isoforms replace the long isoforms bound to C1 and C2 in the quiescent liver. Therefore the basal promoter activity requires an interaction between the long C/EBP isoforms bound to C1 and C2 whereas the inflammation-induced down-regulation results from the loss of interaction between the cytokine-induced, short C/EBP isoforms. PMID- 14530445 TI - A molecular dynamics simulation study of oriented DNA with polyamine and sodium counterions: diffusion and averaged binding of water and cations. AB - Four different molecular dynamics (MD) simulations have been performed for ordered DNA decamers, d(5'-ATGCAGTCAG)*d(5'-TGACTGCATC). The counterions were the two natural polyamines spermidine3+ (Spd3+) and putrescine2+ (Put2+), the synthetic polyamine diaminopropane2+ (DAP2+) and Na+. The simulation set-up corresponds to an infinite array of parallel DNA mimicking the state in oriented DNA fibers or crystals. This work describes general properties of polyamine and Na+ binding to DNA. Simulated diffusion coefficients show satisfactory agreement with experimental NMR diffusion data of comparable systems. The interaction of the polyamines with DNA is dynamic in character and the cations mostly form short lived contacts with the electronegative binding sites of DNA. Polyamines, Na+ and water interact most frequently with the charged phosphate atoms with preference for association from the minor groove side with O1P over O2P. There is a strong anti-correlation in the cation binding to the electronegative groups of DNA, i.e. the presence of a cation near one of the DNA sites repels other cations from binding to this and to the other sites separated by <7.5 A from each other. In contrast to the other polyamines, DAP2+ is able to form 'bridges' connecting neighboring phosphate groups along the DNA strand. A small fraction of DAP2+ and Put2+ can be found in the major grooves, while Spd3+ is absent there. The results of the MD simulations reveal principal differences in the polyamine-DNA interactions between the natural (Spd3+, Put2+ and spermine4+) and synthetic (DAP2+) polyamines. PMID- 14530446 TI - RNA cleaving '10-23' DNAzymes with enhanced stability and activity. AB - '10-23' DNAzymes can be used to cleave any target RNA in a sequence-specific manner. For applications in vivo, they have to be stabilised against nucleolytic attack by the introduction of modified nucleotides without obstructing cleavage activity. In this study, we optimise the design of a DNAzyme targeting the 5'-non translated region of the human rhinovirus 14, a common cold virus, with regard to its kinetic properties and its stability against nucleases. We compare a large number of DNAzymes against the same target site that are stabilised by the use of a 3'-3'-inverted thymidine, phosphorothioate linkages, 2'-O-methyl RNA and locked nucleic acids, respectively. Both cleavage activity and nuclease stability were significantly enhanced by optimisation of arm length and content of modified nucleotides. Furthermore, we introduced modified nucleotides into the catalytic core to enhance stability against endonucleolytic degradation without abolishing catalytic activity. Our findings enabled us to establish a design for DNAzymes containing nucleotide modifications both in the binding arms and in the catalytic core, yielding a species with up to 10-fold enhanced activity and significantly elevated stability against nucleolytic cleavage. When transferring the design to a DNAzyme against a different target, only a slight modification was necessary to retain activity. PMID- 14530447 TI - pi protein- and ATP-dependent transitions from 'closed' to 'open' complexes at the gamma ori of plasmid R6K. AB - R6K-encoded pi protein can bind to the seven, 22 bp tandem iterons of the gamma origin. In this work, we use a variant of pi, His-pi.F107S, that is hyperactive in replication. In vitro, His-pi.F107S-dependent local DNA melting (open complex formation) occurs in the absence of host proteins (IHF/HU or DnaA) and it is positioned in the A + T-rich region adjacent to iterons. Experiments described here examine the effects of ATP, Mg2+ and temperature on the opening reaction. We show that the opening of the gamma origin can occur in the presence of ATP as well as AMP-PCP (a non-hydrolyzable ATP analog). This suggests that, for gamma origin, ATP hydrolysis may be unnecessary for open complex formation facilitated by His-pi.F107S. In the absence of ATP or Mg2+, His-pi.F107S yielded data suggestive of distortions in the iteron attributable to DNA bending rather than DNA melting. Our findings also demonstrate that ATP and pi stimulate open complex formation over a wide range of temperatures, but not at 0 degrees C. These and other results indicate that ATP and/or Mg2+ are not needed for His-pi.F107S binding to iterons and that ATP effects an allosteric change in the protein bound to gamma origin. PMID- 14530448 TI - Human RNA polymerase II is partially blocked by DNA adducts derived from tumorigenic benzo[c]phenanthrene diol epoxides: relating biological consequences to conformational preferences. AB - Environmental polycyclic aromatic hydrocarbons (PAHs) are metabolically activated to diol epoxides that can react with DNA, resulting in covalent modifications to the bases. The (+)- and (-)-3,4-dihydroxy-1,2-epoxy-1,2,3,4-tetrahydro benzo[c]phenanthrene (anti-BPhDE) isomers are diol epoxide metabolites of the PAH benzo[c]phenanthrene (BPh). These enantiomers readily react with DNA at the N6 position of adenine, forming bulky (+)-1R- or (-)-1S-trans-anti-[BPh]-N6-dA adducts. Transcription-coupled nucleotide excision repair clears such bulky adducts from cellular DNA, presumably in response to RNA polymerase transcription complexes that stall at the bulky lesions. Little is known about the effects of [BPh]-N6-dA lesions on RNA polymerase II, hence, the behavior of human RNA polymerase II was examined at these adducts. A site-specific, stereochemically pure [BPh]-N6-dA adduct was positioned on the transcribed or non-transcribed strand of a DNA template with a suitable promoter for RNA polymerase II located upstream from the lesion. Transcription reactions were then carried out with HeLa nuclear extract. Each [BPh]-dA isomer strongly impeded human RNA polymerase II progression when it was located on the transcribed strand; however, a small but significant degree of lesion bypass occurred, and the extent of polymerase blockage and bypass was dependent on the stereochemistry of the adduct. Molecular modeling of the lesions supports the idea that each adduct can exist in two orientations within the polymerase active site, one that permits nucleotide incorporation and another that blocks the RNA polymerase nucleotide entry channel, thus preventing base incorporation and causing the polymerase to stall or arrest. PMID- 14530449 TI - Distance preferences in the arrangement of binding motifs and hierarchical levels in organization of transcription regulatory information. AB - We explored distance preferences in the arrangement of binding motifs for five transcription factors (Bicoid, Kruppel, Hunchback, Knirps and Caudal) in a large set of Drosophila cis-regulatory modules (CRMs). Analysis of non-overlapping binding motifs revealed the presence of periodic signals specific to particular combinations of binding motifs. The most striking periodic signals (10 bp for Bicoid and 11 bp for Hunchback) suggest preferential positioning of some binding site combinations on the same side of the DNA helix. We also analyzed distance preferences in arrangements of highly correlated overlapping binding motifs, such as Bicoid and Kruppel. Based on the distance analysis, we extracted preferential binding site arrangements and proposed models for potential composite elements (CEs) and antagonistic motif pairs involved in the function of developmental CRMs. Our results suggest that there are distinct hierarchical levels in the organization of transcription regulatory information. We discuss the role of the hierarchy in understanding transcriptional regulation and in detection of transcription regulatory regions in genomes. PMID- 14530450 TI - Dissimilar mispair-recognition spectra of Arabidopsis DNA-mismatch-repair proteins MSH2*MSH6 (MutSalpha) and MSH2*MSH7 (MutSgamma). AB - Besides orthologs of other eukaryotic mismatch-repair (MMR) proteins, plants encode MSH7, a paralog of MSH6. The Arabidopsis thaliana recognition heterodimers AtMSH2*MSH6 (AtMutSalpha) and AtMSH2*MSH3 (AtMutSbeta) were previously found to bind the same subsets of mismatches as their counterparts in other eukaryotes- respectively, base-base mismatches and single extra nucleotides, loopouts of extra nucleotides (one or more) only--but AtMSH2*MSH7 (AtMutSgamma) bound well only to a G/T mismatch. To test hypotheses that MSH7 might be specialized for G/T, or for base mismatches in 5-methylcytosine contexts, we compared binding of AtMutSalpha and AtMutSgamma to a series of mismatched DNA oligoduplexes, relative to their (roughly similar) binding to G/T DNA. AtMutSgamma bound G/G, G/A, A/A and especially C/A mispairs as well or better than G/T, in contrast to MutSalpha, for which G/T was clearly the best base mismatch. The presence of 5 methylcytosine adjacent to or in a mispair generally lowered binding by both heterodimers, with no systematic difference between the two. Alignment of protein sequences reveals the absence in MSH7 of the clamp domains that in bacterial MutS proteins--and by inference MSH6 proteins--non-specifically bind the backbone of mismatched DNA, raising new questions as to how clamp domains enhance mismatch recognition. Plants must rigorously suppress mutation during mitotic division of meristematic cells that eventually give rise to gametes and may also use MMR proteins to antagonize homeologous recombination. The MSH6 versus MSH7 divergence may reflect specializations for particular mismatches and/or sequence contexts, so as to increase both DNA-replication and meiotic-recombination fidelity, or dedication of MSH6 to the former and MSH7 to the latter, consistent with genetic evidence from wheat. PMID- 14530451 TI - Solvation change and ion release during aminoacylation by aminoacyl-tRNA synthetases. AB - Discrimination between cognate and non-cognate tRNAs by aminoacyl-tRNA synthetases occurs at several steps of the aminoacylation pathway. We have measured changes of solvation and counter-ion distribution at various steps of the aminoacylation pathway of glutamyl- and glutaminyl-tRNA synthetases. The decrease in the association constant with increasing KCl concentration is relatively small for cognate tRNA binding when compared to known DNA-protein interactions. The electro-neutral nature of the tRNA binding domain may be largely responsible for this low ion release stoichiometry, suggesting that a relatively large electrostatic component of the DNA-protein interaction free energy may have evolved for other purposes, such as, target search. Little change in solvation upon tRNA binding is seen. Non-cognate tRNA binding actually increases with increasing KCl concentration indicating that charge repulsion may be a significant component of binding free energy. Thus, electrostatic interactions may have been used to discriminate between cognate and non-cognate tRNAs in the binding step. The catalytic constant of glutaminyl-tRNA synthetase increases with increasing osmotic pressure indicating a water release of 8.4 +/- 1.4 mol/mol in the transition state, whereas little change is seen in the case of glutamyl-tRNA synthetase. We propose that the significant amount of water release in the transition state, in the case of glutaminyl-tRNA synthetase, is due to additional contact of the protein with the tRNA in the transition state. PMID- 14530452 TI - A compositional segmentation of the human mitochondrial genome is related to heterogeneities in the guanine mutation rate. AB - We applied a hidden Markov model segmentation method to the human mitochondrial genome to identify patterns in the sequence, to compare these patterns to the gene structure of mtDNA and to see whether these patterns reveal additional characteristics important for our understanding of genome evolution, structure and function. Our analysis identified three segmentation categories based upon the sequence transition probabilities. Category 2 segments corresponded to the tRNA and rRNA genes, with a greater strand-symmetry in these segments. Category 1 and 3 segments covered the protein- coding genes and almost all of the non-coding D-loop. Compared to category 1, the mtDNA segments assigned to category 3 had much lower guanine abundance. A comparison to two independent databases of mitochondrial mutations and polymorphisms showed that the high substitution rate of guanine in human mtDNA is largest in the category 3 segments. Analysis of synonymous mutations showed the same pattern. This suggests that this heterogeneity in the mutation rate is partly independent of respiratory chain function and is a direct property of the genome sequence itself. This has important implications for our understanding of mtDNA evolution and its use as a 'molecular clock' to determine the rate of population and species divergence. PMID- 14530453 TI - Detection of regulatory circuits by integrating the cellular networks of protein protein interactions and transcription regulation. AB - The post-genomic era is marked by huge amounts of data generated by large-scale functional genomic and proteomic experiments. A major challenge is to integrate the various types of genome-scale information in order to reveal the intra- and inter- relationships between genes and proteins that constitute a living cell. Here we present a novel application of classical graph algorithms to integrate the cellular networks of protein-protein interactions and transcription regulation. We demonstrate how integration of these two networks enables the discovery of simple as well as complex regulatory circuits that involve both protein-protein and protein-DNA interactions. These circuits may serve for positive or negative feedback mechanisms. By applying our approach to data from the yeast Saccharomyces cerevisiae, we were able to identify known simple and complex regulatory circuits and to discover many putative circuits, whose biological relevance has been assessed using various types of experimental data. The newly identified relations provide new insight into the processes that take place in the cell, insight that could not be gained by analyzing each type of data independently. The computational scheme that we propose may be used to integrate additional functional genomic and proteomic data and to reveal other types of relations, in yeast as well as in higher organisms. PMID- 14530454 TI - Sequential DEXAS: a method for obtaining DNA sequences from genomic DNA and blood in one reaction. AB - Sequential DEXAS (direct exponential amplification and sequencing), a one step amplification and sequencing procedure that allows accurate, inexpensive and rapid DNA sequence determination directly from genomic DNA, is described. This method relies on the simultaneous use of two DNA polymerases that differ both in their ability to incorporate dideoxynucleotides and in the time at which they are activated during the reaction. One enzyme, which incorporates deoxynucleotides and performs amplification of the target DNA sequence, is supplied in an active state whereas the other enzyme, which incorporates dideoxynucleotides and performs the sequencing reaction, is supplied in an inactive state but becomes activated by a temperature step during the thermocycling. Thus, in the initial stage of the reaction, target amplification occurs, while in the second stage the sequencing reaction takes place. We show that Sequential DEXAS yields high quality sequencing results directly from genomic DNA as well as directly from human blood without any prior isolation or purification of DNA. PMID- 14530455 TI - Standardized determination of real-time PCR efficiency from a single reaction set up. AB - We propose a computing method for the estimation of real-time PCR amplification efficiency. It is based on a statistic delimitation of the beginning of exponentially behaving observations in real-time PCR kinetics. PCR ground fluorescence phase, non-exponential and plateau phase were excluded from the calculation process by separate mathematical algorithms. We validated the method on experimental data on multiple targets obtained on the LightCycler platform. The developed method yields results of higher accuracy than the currently used method of serial dilutions for amplification efficiency estimation. The single reaction set-up estimation is sensitive to differences in starting concentrations of the target sequence in samples. Furthermore, it resists the subjective influence of researchers, and the estimation can therefore be fully instrumentalized. PMID- 14530456 TI - A novel real-time quantitative PCR method using attached universal template probe. AB - A novel real-time quantitative polymerase chain reaction (PCR) method using an attached universal template (UT) probe is described. The UT is an approximately 20 base attachment to the 5' end of a PCR primer, and it can hybridize with a complementary TaqMan probe. One of the advantages of this method is that different target DNA sequences can be detected employing the same UT probe, which substantially reduces the cost of real-time PCR set-up. In addition, this method could be used for simultaneous detection using a 6-carboxy-fluorescein-labeled UT probe for the target gene and a 5-hexachloro-fluorescein-labeled UT probe for the reference gene in a multiplex reaction. Moreover, the requirement of target DNA length for UT-PCR analysis is relatively flexible, and it could be as short as 56 bp in this report, suggesting the possibility of detecting target DNA from partially degraded samples. The UT-PCR system with degenerate primers could also be designed to screen homologous genes. Taken together, our results suggest that the UT-PCR technique is efficient, reliable, inexpensive and less labor-intensive for quantitative PCR analysis. PMID- 14530457 TI - Pentaprobe: a comprehensive sequence for the one-step detection of DNA-binding activities. AB - The rapid increase in the number of novel proteins identified in genome projects necessitates simple and rapid methods for assigning function. We describe a strategy for determining whether novel proteins possess typical sequence-specific DNA-binding activity. Many proteins bind recognition sequences of 5 bp or less. Given that there are 4(5) possible 5 bp sites, one might expect the length of sequence required to cover all possibilities would be 4(5) x 5 or 5120 nt. But by allowing overlaps, utilising both strands and using a computer algorithm to generate the minimum sequence, we find the length required is only 516 base pairs. We generated this sequence as six overlapping double-stranded oligonucleotides, termed pentaprobe, and used it in gel retardation experiments to assess DNA binding by both known and putative DNA-binding proteins from several protein families. We have confirmed binding by the zinc finger proteins BKLF, Eos and Pegasus, the Ets domain protein PU.1 and the treble clef N- and C terminal fingers of GATA-1. We also showed that the N-terminal zinc finger domain of FOG-1 does not behave as a typical DNA-binding domain. Our results suggest that pentaprobe, and related sequences such as hexaprobe, represent useful tools for probing protein function. PMID- 14530458 TI - Sequence-specific fluorescent labeling of double-stranded DNA observed at the single molecule level. AB - Fluorescent labeling of a short sequence of double-stranded DNA (dsDNA) was achieved by ligating a labeled dsDNA fragment to a stem-loop triplex forming oligonucleotide (TFO). After the TFO has wound around the target sequence by ligand-induced triple helix formation, its extremities hybridize to each other, leaving a dangling single-stranded sequence, which is then ligated to a fluorescent dsDNA fragment using T4 DNA ligase. A non-repeated 15 bp sequence present on lambda DNA was labeled and visualized by fluorescence microscopy after DNA combing. The label was found to be attached at a specific position located at 4.2 +/- 0.5 kb from one end of the molecule, in agreement with the location of the target sequence for triple helix formation (4.4 kb from one end). In addition, an alternative combing process was noticed in which a DNA molecule becomes attached to the combing slide from the label rather than from one of its ends. The method described herein provides a new tool for the detection of very short sequences of dsDNA and offers various perspectives in the micromanipulation of single DNA molecules. PMID- 14530459 TI - Single-molecule analysis of the hypermutable tetranucleotide repeat locus D21S1245 through sperm genotyping: a heterogeneous pattern of mutation but no clear male age effect. AB - Single molecule genotyping of the hypermutable microsatellite locus D21S1245 was used for studying how the rate and pattern of mutation varied between alleles and different age groups. In total, 203 mutation events were scored from the genotyping of DNA corresponding to an estimated 8623 sperm cells from eight different men. Allele-specific mutation rates ranged from 0.007 to 0.052, a heterogeneity related in part to variation in the mutation rate among three allelic lineages identified after allele sequencing. Alleles from these lineages differed in the overall repeat structure of this complex microsatellite locus. Also, the pattern of mutation varied between lineages in that they differed in the relative proportions of expansion and contraction mutations. Surprisingly, a group of four men aged 18-23 years showed a higher mean mutation rate than a group of four men aged 48-56 years. To some extent this age difference can probably be explained by a bias in the distribution of alleles from the three allelic lineages among the age groups. However, the absence of a clear male age effect is at odds with the idea of an increasing male mutation rate with age, which is thought to arise from the continuous replication of germline cells throughout adulthood. PMID- 14530465 TI - Orbital scintigraphy with the somatostatin receptor tracer 99mTc-P829 in patients with Graves' disease. AB - Receptors for somatostatin (SST) (SSTR) are expressed on various tumor cells as well as on activated lymphocytes. Previous data have shown that (99m)Tc-P829 binds with high affinity to many different types of tumor cells as well as to leukocytes via the human hSSTR2, hSSTR3, and hSSTR5 target receptors. Consequently, (99m)Tc-P829 was successfully introduced as a peptide tracer for tumor imaging. In this study, we evaluated the orbital uptake of (99m)Tc-P829 in patients with active and inactive thyroid-associated orbitopathy (TAO), accompanied by lymphocyte infiltration in the acute stage and by muscle fibrosis in the chronic stage of the disease. METHODS: To evaluate its clinical usefulness in Graves' disease, (99m)Tc-P829 scintigraphy (approximately equal to 740 MBq) was performed in 44 patients with TAO (median duration, 19 mo; range, 1-360 mo). The clinical activity of the orbital disease was graded by the NOSPECS (no signs or symptoms; only signs, no symptoms; signs only; proptosis; eye muscle involvement; corneal involvement; sight visual acuity reduction) classification of the American Thyroid Association, the clinical activity score (CAS), and the superonasal index (SNI). SPECT (360 degrees ) and planar studies were completed within 3 h after injection. Orbital (O) regions of interest (ROIs) were compared with temporoparietal and occipital (OCC) ROIs. Orbital uptake ratios in Graves' disease were compared with data obtained from lung cancer patients with no eye disease (n = 22). RESULTS: Overall, (99m)Tc-P829 biokinetics were the same in Graves' disease patients as in lung cancer patients, showing a rapid blood clearance and visualization of the facial bones within minutes of injection. In all control patients, the orbit appeared as a "cold area," whereas visual orbital accumulation of (99m)Tc-P829 was found in patients with active TAO (O/OCC ratios: 1.26 +/- 0.04 vs. 1.69 +/- 0.04; P < 0.01, respectively). Patients with active eye disease (n = 25) presented with an increased orbital uptake of (99m)Tc-P829 compared with patients with inactive disease (n = 19; O/OCC ratio: 1.12 +/- 0.05; P < 0.01). A statistically significant correlation was found between CAS and the orbital uptake (O/OCC ratio) values (r = 0.90), whereas no correlation could be documented regarding the NOSPECS classification as well as the SNI. CONCLUSION: In TAO, (99m)Tc-P829 yields high orbital binding with good clinical correlation. The better image quality due to the high energy of technetium, the lower radiation dose for patients and personnel, and the short acquisition protocol favor SSTR scintigraphy with (99m)Tc-P829 over (111)In-labeled compounds. The in house availability of the radiotracer and cost-effectiveness are further advantages. PMID- 14530466 TI - Evaluating the clinical effectiveness of 90Y-SMT 487 in patients with neuroendocrine tumors. AB - Because of the presence of cell membrane somatostatin receptors (SSTRs), many neuroendocrine tumors will bind analogs of somatostatin. (90)Y Dodecanetetraacetic acid-Phe1-Tyr3-octreotide (SMT 487) is an SSTR radiopharmaceutical currently under investigation as a therapeutic option for neuroendocrine tumors. Although there are a variety of methods for evaluating response to a given cancer therapy, an important indicator of success is the impact on the clinical status of the patient. The purpose of this work was to develop a semiquantitative method and assess the clinical effectiveness of (90)Y SMT 487 therapy in patients with neuroendocrine tumors. METHODS: A scoring system was developed to evaluate clinical response that included the following parameters: weight, health status score (determined by the patient), Karnofsky score, and tumor-related symptoms. RESULTS: We applied this scoring system to 21 patients who had completed 3 cycles of therapy with (90)Y-SMT 487. Fourteen of the 21 showed a favorable clinical response, whereas 5 were clinically stable after treatment and 2 showed evidence of clinical progression. There was also a significant reduction in the amount of octreotide being used after completion of (90)Y-SMT 487 therapy in the 20 patients who were on this medication. CONCLUSION: Using this scoring method, (90)Y-SMT 487 appears effective in improving the clinical status of patients with (111)In-pentetreotide-positive neuroendocrine tumors. PMID- 14530467 TI - Staging of regional lymph nodes in melanoma patients by means of 99mTc-MIBI scintigraphy. AB - Most first relapses in patients with melanoma occur in regional lymph node basins. Such lesions are frequently diagnosed clinically during the first 2 y of follow-up. In the last few years, our group has been studying the usefulness of (99m)Tc-methoxyisobutylisonitrile (MIBI) scintigraphy in the evaluation of recurrent melanoma lesions. The aim of the present study was to prospectively evaluate the clinical value of (99m)Tc-MIBI scintigraphy in the diagnosis of subclinical nodal metastases. METHODS: We included 66 patients within 3 mo of melanoma diagnosis, with Breslow thickness > 1.0 mm, all treated with wide local excision of the primary lesion. When (99m)Tc-MIBI scanning was performed, 49 of them did not have evidence of nodal disease, and 17 had clinically questionable regional lymph node lesions. Planar images of lymph node regions were acquired 10 min after injection, using a dose of 740-1,110 MBq and a large-field-of-view gamma camera equipped with a low-energy high-resolution collimator. Scan findings were confirmed by pathology or by clinical follow-up (median, 35 mo). RESULTS: Thirty of 33 patients with regional lymph node metastases received a correct diagnosis, 14 with palpable lesions and 16 with nonpalpable lesions. In 3 cases that were initially (99m)Tc-MIBI negative, nodal metastases were found during follow-up. The following diagnostic values were calculated: sensitivity, 0.91 (95% confidence interval [CI], 0.75-0.98); specificity, 0.85 (95% CI, 0.67-0.94); likelihood ratio of a positive test, 6.0 (95% CI, 2.7-13.5); and likelihood ratio of a negative test, 0.11 (95% CI, 0.036-0.32). CONCLUSION: (99m)Tc-MIBI scanning may have a secondary role in the staging of regional lymph nodes in patients with clinically localized melanoma who are not good candidates for sentinel node biopsy. PMID- 14530468 TI - 99mTc-MIBI SPECT in distinguishing neoplastic from nonneoplastic intracerebral hematoma. AB - Distinguishing neoplastic from nonneoplastic intracerebral hematoma has great clinical relevance for the appropriate management of patients. Imaging is not always able to clearly identify a tumor-related intraparenchymal cerebral hemorrhage (ICH), especially in the acute phase, the diagnosis being frequently based on evolution patterns. The aim of this study was to test the value of (99m)Tc-methoxyisobutylisonitrile ((99m)Tc-MIBI) SPECT as a noninvasive diagnostic tool in early diagnosis of hemorrhagic brain neoplasm. METHODS: We prospectively studied 29 patients harboring a nontraumatic acute onset of clinical deterioration caused by ICH with atypical clinical or neuroradiologic features. All patients underwent (99m)Tc-MIBI SPECT within 48 h from the clinical onset. Early and delayed images were obtained. Both visual and semiquantitative analyses were performed. The (99m)Tc-MIBI index was obtained from both early and delayed images and the retention index was calculated. RESULTS: In 19 patients (65.5%), a nonneoplastic hemorrhage (15 vascular degenerative diseases, 2 cavernous angiomas, 1 thrombosed middle cerebral artery giant aneurysm, and 1 sinus rectus thrombosis) was diagnosed by clinical and neuroradiologic follow-up or open surgery. In 10 patients (34.5%), a neoplastic hemorrhage (6 metastases, 2 glioblastomas multiforme, 1 ependymoma, and 1 intracranial angioblastic meningioma) was diagnosed by direct histologic typing (open surgery or stereotactic biopsy). In all neoplasm-related hemorrhages, a focal increased tracer uptake was observed in the area of the lesion, whereas no focal increased tracer uptake was noted in all nonneoplastic hematomas. A wide cutoff in the early ratio between neoplastic and nonneoplastic hemorrhages was found. Moreover, a statistically significant difference was found in the delayed ratio (P < 0.01) and the retention index (P < 0.05) between the 2 groups. CONCLUSION: Our data suggest that (99m)Tc-MIBI SPECT could play a role in the early noninvasive diagnostic work-up of hemorrhagic brain lesions, allowing a clear differentiation between neoplastic and nonneoplastic ICHs. The high availability and low cost of this nuclear medicine technique can be considered additional advantages. PMID- 14530469 TI - Reliability of captopril renography in patients under chronic therapy with angiotensin II (AT1) receptor antagonists. AB - Captopril renography is a reliable, widely used test for the functional diagnosis of renovascular hypertension. Well-recognized drawbacks of the procedure include reduced accuracy in patients with bilateral disease or renal impairment as well as the possible interference from concurrent antihypertensive medication (diuretics, beta-blockers, angiotensin-converting enzyme inhibitors, calcium channel blockers). Currently, no data exist regarding the reliability of captopril renography in patients with renovascular hypertension evaluated while they are under chronic treatment with angiotensin II (AT1) receptor antagonists (Sartans). Moreover, the renographic response of the kidney with renal artery stenosis to prolonged therapy with angiotensin II receptor antagonists has not yet been evaluated. METHODS: We investigated the diagnostic effectiveness of (99m)Tc-mercaptoacetyltriglycine captopril renography performed after acute addition of 25 mg of captopril to the daily dose of AT1 receptor antagonist in 13 patients with unilateral renal artery stenosis and subsequent evidence of renovascular hypertension, based on short-term (3 mo) blood pressure outcome after revascularization. The renographic evaluation was first performed after ingestion of the daily therapy of angiotensin II receptor antagonist alone (Sartan renography) and was repeated within 7 d after the acute addition of 25 mg of captopril to chronic treatment with angiotensin II receptor antagonist (captopril-Sartan renography). A cohort of 13 patients with a final diagnosis of essential hypertension was chosen as the control subjects. RESULTS: Twelve of 13 patients were correctly detected by captopril-Sartan renography (92% sensitivity), and 3 subjects were also identified without the addition of captopril. Adding captopril to Sartan therapy resulted in a slight reduction in mean arterial blood pressure, while significant side effects were never observed. No false-positive results were found in the 13 patients with essential hypertension. CONCLUSION: We conclude that performing captopril renography with the acute addition of 25 mg of captopril to the chronic monotherapy with Sartans has the same diagnostic effectiveness as performing captopril renography alone. Interrupting the vasoactive action of angiotensin II alone on the efferent glomerular arteries, which can also be selectively achieved by chronic administration of angiotensin II receptor antagonists, does not fully explain the effectiveness of captopril renography in detecting renovascular hypertensive patients. PMID- 14530470 TI - Visualization of interscapular brown adipose tissue using (99m)Tc-tetrofosmin in pediatric patients. AB - Brown adipose tissue (BAT) is a site of nonshivering thermogenesis in mammals. The mitochondria of BAT operate in an uncoupled mode and increase fatty acid oxidation to produce heat at birth. Thus, the BAT of human infants and children contains more active mitochondria than that of adults. We surmised that because (99m)Tc-tetrofosmin can be absorbed by functional mitochondria in the myocardium and in tumor cells, it could reveal mitochondrial function in BAT. METHODS: Between January 1999 and December 2002, we retrospectively analyzed 385 consecutive studies of (99m)Tc-tetrofosmin uptake in pediatric patients with cardiac disorders. All patients with symmetric (99m)Tc-tetrofosmin accumulation within the neck and shoulder region according to planar images were selected, and the features of the uptake were analyzed. RESULTS: Increased symmetric (99m)Tc tetrofosmin uptake in the interscapular BAT was a typical profile of 65 of the 385 patients (17%). The frequency of (99m)Tc-tetrofosmin uptake was significantly higher in winter than in spring or summer (P < 0.05) and prominent in newborns. The frequency peaked between 0 and 2 y of age and then declined with age. CONCLUSION: Gamma-camera imaging with (99m)Tc-tetrofosmin can reveal interscapular BAT distribution in infants and children in terms of mitochondrial activity. PMID- 14530471 TI - Inflammatory bowel disease activity assessment with biologic markers and 99mTc WBC scintigraphy: are there different trends in ileitis versus colitis? AB - To evaluate whether scintigraphy with (99m)Tc-labeled white blood cells (WBC) can assess the intensity of bowel inflammation, a large dataset of laboratory values and clinical activity indices was correlated with (99m)Tc-WBC scintigraphy in children with Crohn's disease (CD), ulcerative colitis (UC), and miscellaneous colitis (MC). Also evaluated was whether stratification of children with CD as ileitis versus colitis results in different correlation coefficients for laboratory values versus (99m)Tc-WBC scintigraphy. METHODS: Over a 6-y period, 313 (99m)Tc-WBC studies were performed. A dataset of 2,714 laboratory values is available for analysis. RESULTS: There is a positive correlation between the erythrocyte sedimentation rate (ESR) and large bowel uptake of (99m)Tc-WBC (P < 0.05) and a negative trend with small bowel uptake of (99m)Tc-WBC in children with CD. Similarly, there is a correlation between WBC counts and scintigraphy in most segments of the large bowel and a negative correlation with the small bowel (R = -0.32, P = 0.01) in children with CD. There is a correlation between platelets and (99m)Tc-WBC in children with CD or UC. There is no correlation between the ESR and (99m)Tc-WBC in children with UC or MC. Many clinical activity indices correlate (P < 0.001) with (99m)Tc-WBC in children with CD, but none correlates in children with UC. Numerous laboratory values correlate with each other. There is a negative correlation between protein, albumin, hemoglobin, and hematocrit versus (99m)Tc-WBC scintigraphy in children with CD. In children with UC, there is a negative correlation between hemoglobin and hematocrit versus (99m)Tc-WBC. CONCLUSION: (99m)Tc-WBC scintigraphy, ESR, and WBC counts are good indicators of the inflammatory activity in CD if the inflammation is limited to the large bowel. There is a trend toward an inverse relationship when the inflammation is limited to the small bowel; thus, scintigraphy and the aforementioned markers may be of limited value. This report also demonstrates that (99m)Tc-WBC scintigraphy correlates with clinical activity indices in CD and with numerous biologic markers. In children with UC, scintigraphy with (99m)Tc WBC and most laboratory markers are of limited value in assessing disease activity. PMID- 14530472 TI - Simple quantification of skeletal muscle glucose utilization by static 18F-FDG PET. AB - Skeletal muscle glucose utilization (SMGU) can be measured by dynamic PET imaging with (18)F-FDG to characterize insulin resistance. The aim of this study was to determine the validity of simple methods to quantify SMGU by static PET imaging. METHODS: Ten patients underwent dynamic (18)F-FDG PET of the femoral region during hyperinsulinemic euglycemic clamping. SMGU was determined by Patlak graphical analysis using data from dynamic imaging with frequent arterial blood sampling. Standardized uptake values (SUVs) were calculated at 45 and 55 min after tracer injection. Skeletal muscle-to-background ratio (SM/B ratio), tissue count divided by venous plasma activity, was also computed at 45 and 55 min. These simple indices were compared by linear regression with the SMGU measured as above, and an estimated SMGU was obtained using the regression equation thus generated, together with a simple index. RESULTS: SMGU was highly correlated with SUVs (r = 0.941 at 45 min, r = 0.951 at 55 min) and SM/B ratios (r = 0.968 at 45 min, r = 0.984 at 55 min). Although SMGU was almost proportional to SM/B ratios, the y-intercepts of the regression lines for SUVs significantly differed from zero. The residual in estimating SMGU using the regression equation was marginally smaller for SM/B ratios than for SUVs and for indices at 55 min than at 45 min, but these differences did not reach statistical significance. Correction for plasma glucose level slightly elevated the correlation coefficients between SMGU and these simple indices. CONCLUSION: It is proposed that the simple quantitative indices, SUV and the SM/B ratio, are reliable indicators of SMGU during hyperinsulinemic euglycemic clamping. Static imaging with or without a single venous blood sampling may therefore be able to replace dynamic imaging with frequent arterial blood sampling, offering substantially greater convenience in evaluating insulin resistance. PMID- 14530473 TI - Aging-related changes in cardiac sympathetic function in humans, assessed by 6 18F-fluorodopamine PET scanning. AB - Sympathetic nerves play key roles in cardiac physiology and aging-related cardiovascular diseases. This study examined the effects of normal human aging on cardiac sympathetic innervation and function, including the neuronal uptake of catecholamines (uptake 1) via the cell membrane norepinephrine transporter. METHODS: Thirty-three healthy volunteers, 17 under 40 and 16 over 50 y old, underwent thoracic PET scanning after injection of the sympathoneural imaging agent 6-(18)F-fluorodopamine. Myocardial perfusion was estimated by (13)NH(3) scanning, and arterial blood was sampled for levels of 6-(18)F-fluorodopamine and 6-(18)F-fluorodopamine-derived radioactivity. RESULTS: The older group had more myocardial 6-(18)F-fluorodopamine-derived radioactivity than did the younger group. Myocardial perfusion was also greater in the older group, and arterial blood levels of 6-(18)F-fluorodopamine were also higher. After adjustment for delivery of the tracer, the estimated level of myocardial extraction of 6-(18)F fluorodopamine was lower in the older group (48%) than in the younger group (74%) (P = 0.02). CONCLUSION: Cardiac uptake 1 activity decreases with normal human aging. PMID- 14530474 TI - Efficacy of carvedilol treatment on cardiac function and cardiac sympathetic nerve activity in patients with dilated cardiomyopathy: comparison with metoprolol therapy. AB - Carvedilol and metoprolol have been reported to be effective in the treatment of patients with chronic heart failure. However, to our knowledge, there have been no studies comparing the effects of the 2 drugs on cardiac function, including cardiac sympathetic nerve activity. METHODS: We compared 15 patients with dilated cardiomyopathy (DCM) who were receiving carvedilol (group A) with 15 patients with DCM who were receiving metoprolol (group B). Before and after 1 y of treatment, cardiac (123)I-metaiodobenzylguanidine ((123)I-MIBG) uptake was assessed using the total defect score (TDS) and the heart-to-mediastinum (H/M) activity ratio from the delayed images. The New York Heart Association (NYHA) class and echocardiographic left ventricular ejection fraction (LVEF) also were assessed. RESULTS: In both groups, the TDS decreased (in group A, from 25 +/- 14 to 16 +/- 14, P < 0.01; in group B, from 27 +/- 9 to 19 +/- 10, P < 0.01), the H/M increased (in group A, from 1.67 +/- 0.31 to 2.01 +/- 0.36, P < 0.01; in group B, from 1.68 +/- 0.21 to 1.93 +/- 0.32, P < 0.01), the LVEF increased (in group A, from 31% +/- 10% to 48% +/- 10%, P < 0.01; in group B, from 28% +/- 9% to 47% +/- 15%, P < 0.01), and the NYHA functional class improved (in group A, from 2.9 +/- 0.3 to 1.7 +/- 0.5, P < 0.01; in group B, from 2.8 +/- 0.6 to 1.7 +/ 0.6, P < 0.01). The change in LVEF was mildly correlated with the change in the TDS in group A (r = 0.41) as well as in group B (r = 0.53). In the patients with a favorable response in the TDS or H/M, the NYHA class improved more than in the patients without a favorable response (P < 0.05). CONCLUSION: Carvedilol treatment can improve cardiac function, symptoms, and cardiac sympathetic nerve activity in patients with DCM to a similar extent as metoprolol treatment. The improvement of cardiac function and symptoms is related to the improvement of cardiac sympathetic nerve activity. PMID- 14530475 TI - Changes in cardiac sympathetic nerve innervation and activity in pathophysiologic transition from typical to end-stage hypertrophic cardiomyopathy. AB - Left ventricular (LV) systolic function in hypertrophic cardiomyopathy (HCM) is usually normal. Late in the disease, however, LV systolic dysfunction and dilatation are recognized. Although abnormalities in cardiac sympathetic nerve activity in patients with HCM have been demonstrated using (123)I metaiodobenzylguanidine (MIBG) myocardial scintigraphy, the changes of cardiac sympathetic nerve activity throughout the clinical course from typical to end stage HCM are unclear. The objective of this study was to evaluate the relationship between abnormalities on (123)I-MIBG myocardial scintigraphy and pathophysiologic changes in patients with HCM. METHODS: We performed (123)I-MIBG scintigraphy on 46 patients with HCM and 18 age-matched control subjects. The patients were categorized into 3 groups: 28 patients with normal LV systolic function (group A), 9 patients with LV systolic dysfunction (group B), and 9 patients with LV systolic dysfunction and dilatation (group C). With planar (123)I-MIBG imaging, the heart-to-mediastinum ratio for early and delayed acquisitions and the washout rate were calculated. With SPECT, polar maps of the LV myocardium were divided into 20 segments. The regional uptake and washout rate were calculated from semiquantitative 20-segment bull's-eye analysis. RESULTS: The early uptake was significantly lower in group C than in the control group (P < 0.01). The washout rate was progressively higher in group A, group B, and group C (P < 0.01). Reduced regional early uptake was found in 2.9 +/- 3.4 (group A), 4.1 +/- 4.7 (group B), and 7.4 +/- 4.3 (group C) segments, respectively. In group C, regional early uptake was significantly reduced, predominantly in the interventricular septal wall, and regional washout rate was increased in the apex and lateral wall. CONCLUSION: These results suggest that cardiac sympathetic nerve abnormalities in patients with HCM may advance with development of LV systolic dysfunction and dilatation and that (123)I-MIBG myocardial scintigraphy may be a useful tool for the evaluation of pathophysiologic changes in HCM. PMID- 14530476 TI - Correlation between cardiac norepinephrine overflow during exercise and cardiac 123I-MIBG uptake in patients with chronic heart failure. AB - Cardiac (123)I-metaiodobenzylguanidine ((123)I-MIBG) uptake is reduced in chronic heart failure, and its reduction is reported to relate to the decrease in exercise capacity. Reduced (123)I-MIBG uptake may predict an inadequately reduced adrenergic drive to the heart during cardiac sympathetic stimulation, including exercise. However, there is little information about the relationship between cardiac (123)I-MIBG uptake at rest and norepinephrine (NE) release during exercise in relation to the exercise capacity in the failing heart. The aim of this study was to examine whether cardiac (123)I-MIBG uptake at rest can predict cardiac sympathetic activity during exercise in patients with chronic heart failure. We determined how cardiac (123)I-MIBG uptake at rest relates to NE overflow from the heart during symptom-limited graded exercise in such patients. METHODS: Twelve patients (mean +/- SD, 52 +/- 12 y) with chronic stable heart failure performed symptom-limited graded exercise tests under catheterizations with a 4-min stage using a supine bicycle ergometer within 2 wk after (123)I-MIBG imaging. NE concentrations in the arterial and coronary sinus blood (NE(A) and NE(CS), respectively) were measured at each exercise stage, and NE overflow was approximated by the difference between NE(CS) and NE(A) (NE(CS-A)). RESULTS: The left ventricular ejection fraction at rest was 47% +/- 16% and peak oxygen uptake was 17.7 +/- 5.1 mL/kg/min. The heart-to-mediastinum uptake ratio of the delayed (123)I-MIBG image (1.00 - 1.72; mean +/- SD, 1.30 +/- 0.19) correlated with NE(CS A) at peak exercise (r = 0.80, P < 0.01) and peak heart rate (r = 0.73, P < 0.01) but not with peak oxygen uptake. CONCLUSION: Cardiac (123)I-MIBG uptake of the delayed image can predict the degree of the increase in adrenergic drive to the heart during sympathetic stimuli induced by exercise in patients with chronic heart failure. PMID- 14530477 TI - Preliminary evaluation of a fuzzy logic-based automatic quantitative analysis in myocardial SPECT. AB - This study validates a new quantitative myocardial perfusion SPECT software. METHODS: The processing starts with the extraction of the morphologic skeleton of the left ventricular myocardium from reconstructed transverse sections. Fuzzy logic is used to decide whether a pixel belongs to the myocardium and any perfusion defect is filled according to a truncated bullet model. The resulting image is partitioned in 18 isovolumetric sectors. Sex-matched normal limits, criteria of abnormality for rest (201)Tl and (99m)Tc-labeled perfusion tracers, reproducibility studies, and detection of coronary artery disease were developed and validated in an overall population of 343 patients. The sex- and tracer matched means and SDs of a normal response were calculated in 93 male and 93 female patients with a <5% likelihood of coronary artery disease. Reproducibility measurements and assignment of different sectors of the myocardium to a specific coronary were performed from data collected in 49 and 60 patients, respectively. The accuracy of the detection of a coronary artery occlusion was assessed in 48 patients who also underwent coronary angiography. RESULTS: The intra- and interoperator reproducibility of the sectorial activity was high with a linear regression coefficient of 0.97 and a SD of the difference measurement at 4.4% and 3.8%, respectively. Overall sensitivity and specificity for the detection of occluded coronary artery were 90% and 80%, respectively. For the detection of left anterior descending, left circumflex, and right artery coronary occlusion, sensitivity was 92%, 75%, and 92.5%, respectively, and specificity was 75%, 78%, and 90%, respectively. CONCLUSION: The new quantitative myocardial perfusion SPECT software appears to be a very helpful program for the objective analysis of perfusion tracer distribution in myocardial SPECT and a very accurate tool in the detection and localization of coronary artery occlusion. PMID- 14530478 TI - Prognostic implications of combined prone and supine acquisitions in patients with equivocal or abnormal supine myocardial perfusion SPECT. AB - Although acquisition of (99m)Tc-sestamibi myocardial perfusion SPECT (MPS) with the patient in the prone position is commonly used to minimize attenuation artifacts, the impact of combined prone and supine imaging on the prognostic evaluation of coronary artery disease (CAD) has not been determined. The prognostic implications of MPS obtained in both prone and supine positions in patients with perfusion defects on supine MPS were evaluated. METHODS: We studied 3,834 patients who were monitored for 24.2 +/- 6.0 mo after rest (201)Tl/stress (99m)Tc-sestamibi MPS acquired during 1994-1995, when prone acquisition was performed only in patients with inferior wall perfusion defects that might represent attenuation or motion artifact. RESULTS: During follow-up, there were 132 hard events (cardiac death or myocardial infarction) and 375 total events (hard events or late myocardial revascularization). Overall, patients who underwent prone and supine acquisitions had similar characteristics to those who underwent supine-only imaging, with the exception of being more commonly male. In multivariable analysis, there were similar independent predictors for hard events and total events; the type of acquisition (prone and supine or supine-only) was not a significant predictor of either of these outcome events. After risk adjustment, the predicted event rates were nearly identical for patients undergoing prone and supine compared with supine-only studies. Both observed and predicted hard event rates of patients with normal prone and supine versus supine only imaging were very low (observed, 0.7%/y and 0.5%/y, respectively; predicted, 1.5% over 24 mo for both). There was no reduction in the higher rates of events associated with abnormal scan results with the combination of prone and supine imaging. CONCLUSION: Patients with inferior wall defects on supine MPS that are not present on prone MPS have a low risk of subsequent cardiac events, similar to that of patients with normal supine-only studies. PMID- 14530479 TI - Implication of prognostically significant negative results on prone SPECT. PMID- 14530480 TI - Reduction of respiratory motion artifacts in PET imaging of lung cancer by respiratory correlated dynamic PET: methodology and comparison with respiratory gated PET. AB - This study proposes a new method to reduce respiratory motion artifacts in PET images of lung cancer. The method is referred to as respiratory-correlated dynamic PET (RCDPET). RCDPET enables the acquisition of 4-dimensional PET data without the need for a respiratory tracking device. In this article, we compare this method with respiratory-gated PET (RGPET). Both methods provide the ability to correct for motion artifacts and more accurately quantitate radiotracer uptake within lung lesions. Both methods were evaluated in phantom studies and 1 patient. METHODS: With RCDPET, data are acquired in consecutive 1-s time frames. A point source attached to a rigid foam block is set on the patient's abdomen and is extended into the camera field of view at the level of the lesion by means of a low-density rod. The position of this source is used to track respiratory motion through the consecutive dynamic frames. Image frames corresponding to a user-selected lesion position within the breathing cycle, in correlation with the point source position, are then identified after scanning. The sinograms of the selected image frames are summed and then reconstructed using iterative reconstruction with segmented attenuation correction. RESULTS: The results from phantom studies with both RGPET and RCDPET were within 10% agreement, for both activity quantitation and image noise levels. In a clinical application, the quantitation of the SUV(max) and the lesion's size showed a 6% and 2% difference, respectively, between RCDPET and RGPET measurements. CONCLUSION: RCDPET can be considered as a comparable, or alternative, method to RGPET in reducing the smearing effects due to respiration and improving quantitation of PET in the thorax. One advantage of RCDPET over RGPET is the ability to retrospectively reconstruct the PET data at any phase or amplitude in the breathing cycle. PMID- 14530481 TI - Radiolabeled neurotensin analog, 99mTc-NT-XI, evaluated in ductal pancreatic adenocarcinoma patients. AB - The study aim was to assess the safety, biodistribution, tissue kinetics, and tumor uptake of the (99m)Tc-labeled neurotensin (NT) analog NT-XI. METHODS: Four patients presenting ductal pancreatic adenocarcinoma were studied with (99m)Tc-NT XI. Patients were followed by scintigraphy up to 4 h and by continued blood and urinary sampling until surgery 18-22 h after injection. Surgical tissue samples were analyzed for radioactivity uptake and NT receptor expression. RESULTS: No side effects were observed on injection of (99m)Tc-NT-XI. Blood biologic half lives alpha and beta were 35 min (range, 17-62 min) and 230 min (range, 107-383 min), respectively. Repeated whole-body scintigraphy performed in 2 patients showed a single exponential decrease of whole-body activity with half-lives of 101 and 232 min. Tracer elimination was mainly renal, with 92% and 98% of activity counted in urine in the first 20 h. Kidney, liver, spleen, and bone marrow activity uptake was observed in all patients. Tumor was not visualized in the first 3 patients but could be localized by tomoscintigraphy in the pancreas head region of patient 4. In vitro tissue analysis showed high expression of NT receptor in the tumor of patient 4, correlated with the highest tumor radioactivity uptake and the highest tumor-to-fat radioactivity ratio. In vitro receptor expression was also positive in a second patient having a tumor characterized by very low cellularity; however, the remaining 2 tumors lacked NT receptor expression. CONCLUSION: Injection of (99m)Tc-NT-XI was well tolerated. The in vivo tumor uptake appeared specific as it was observed in the 1 patient with a pancreatic tumor that expressed high amounts of NT receptor. The results are compatible with preclinical animal results and in favor of further development of radiolabeled NT analogs for diagnosis or therapy of cancer. PMID- 14530482 TI - Quantitative gated PET for the assessment of left ventricular function in small animals. AB - 18F-FDG PET can identify areas of myocardial viability and necrosis and provide useful information on the effectiveness of experimental techniques designed to improve contractile function and myocardial vascularization in small animals. The left ventricular volume (LVV) and left ventricular ejection fraction (LVEF) in normal and diseased rats were measured in vivo using the high-resolution avalanche photodiode (APD) small-animal PET scanner of the Universite de Sherbrooke. The measurements obtained by PET were compared with those obtained by high-resolution echocardiography and with known values obtained from a small, variable-volume cardiac phantom. METHODS: List-mode gated (18)F-FDG PET studies were performed using the APD PET scanner on 30 rats: 11 healthy, 4 under septic shock, and 15 with heart failure induced by ligature of the left coronary artery. PET images were resized to match human-scale pixels and analyzed using a standard clinical cardiac software program. The LVV and LVEF from the same animals were also evaluated by echocardiography. RESULTS: Agreement was excellent between the endocardial volumes determined by PET and the actual volumes of the cardiac phantom (r(2) = 0.96). Agreement between PET and echocardiography for LVV ranged from good in healthy rats (r(2) = 0.89) to fair in diseased rats (r(2) = 0.49). Agreement was fair between LVEF values measured by the 2 methods (r(2) = 0.56). Normal rats had an average LVEF of 83.2% +/- 8.0% using PET and 81.6% +/- 6.0% using echocardiography. In rats with heart failure, LVEF was 54.6% +/- 15.9% using PET and 54.2% +/- 13.3% using echocardiography. CONCLUSION: Both PET and echocardiography clearly differentiated normal rats from rats with heart failure. Echocardiography is fast and convenient, whereas list-mode PET is also able to assess defect size, myocardial viability, and metabolism. PMID- 14530483 TI - Gated PET and ventricular volume. PMID- 14530484 TI - Quantitative 89Zr immuno-PET for in vivo scouting of 90Y-labeled monoclonal antibodies in xenograft-bearing nude mice. AB - Immuno-PET as a scouting procedure before radioimmunotherapy (RIT) aims at the confirmation of tumor targeting and the accurate estimation of radiation dose delivery to both tumor and normal tissues. Immuno-PET with (89)Zr-labeled monoclonal antibodies (mAbs) and (90)Y-mAb RIT might form such a valuable combination. In this study, the biodistribution of (89)Zr-labeled and (88)Y labeled mAb ((88)Y as substitute for (90)Y) was compared and the quantitative imaging performance of (89)Zr immuno-PET was evaluated. METHODS: Chimeric mAb (cmAb) U36, directed against an antigen preferentially expressed in head and neck cancer, was labeled with (89)Zr using the bifunctional chelate N succinyldesferrioxamine B (N-sucDf) and with (88)Y using the bifunctional chelate p-isothiocyanatobenzyl-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (p-SCN-Bz-DOTA). The radioimmunoconjugates were coinjected in xenograft-bearing nude mice, and biodistribution was determined at 3, 24, 48, 72, and 144 h after injection. (89)Zr was evaluated and compared with (18)F in phantom studies to determine linearity, resolution, and recovery coefficients, using a high resolution research tomograph PET scanner. The potential of PET to quantify cmAb U36-N-sucDf-(89)Zr was evaluated by relating image-derived tumor uptake data (noninvasive method) to (89)Zr uptake data derived from excised tumors (invasive method). RESULTS: (89)Zr-N-sucDf-labeled and (88)Y-p-SCN-Bz-DOTA-labeled cmAb U36 showed a highly similar biodistribution, except for sternum and thigh bone at later time points (72 and 144 h after injection). Small differences were found in kidney and liver. Imaging performance of (89)Zr approximates that of (18)F, whereas millimeter-sized (19-154 mg) tumors were visualized in xenograft-bearing mice after injection of cmAb U36-N-sucDf-(89)Zr. After correction for partial volume effects, an excellent correlation was found between image-derived (89)Zr tumor radioactivity and gamma-counter (89)Zr values of excised tumors (R(2) = 0.79). CONCLUSION: The similar biodistribution and the favorable imaging characteristics make (89)Zr a promising candidate for use as a positron-emitting surrogate for (90)Y. PMID- 14530485 TI - Pharmacokinetics and metabolism of 5-125I-iodo-4'-thio-2'-deoxyuridine in rodents. AB - Pharmacokinetics and metabolite analysis of radioiodinated 5-iodo-4'-thio-2' deoxyuridine (ITdU), a potential tumor cell proliferation-imaging agent, were investigated in rodents. METHODS: Blood and urine metabolites of radiolabeled ITdU were analyzed by thin-layer chromatography in rats. Tissue fractionation studies in mice were also performed to investigate the incorporation of (125)I ITdU into DNA. RESULTS: The blood distribution study suggested the influx of (125)I-ITdU into rat peripheral blood cells. The blood metabolism of (125)I-ITdU in vitro was negligible. Plasma analysis at 60 min after injection showed that 50% of the radioactivity was intact (125)I-ITdU. The plasma clearance of (125)I ITdU showed a biexponential function with half-lives of 0.2 and 29.5 min, representing 94.2% and 5.8% of plasma (125)I-ITdU concentration, respectively. (125)I-ITdU was excreted in urine 86% intact at 3.5 h after injection. Extraction of the tissues demonstrated that >70% of the activity in the duodenum, spleen, and thymus was incorporated into DNA at 8 h after injection. On the other hand, <8% of the activity in the liver was in the DNA fraction at that time. CONCLUSION: These data demonstrate that (125)I-ITdU is less susceptible against catabolism and readily incorporated into DNA in proliferating tissues. Therefore, (125)I-ITdU may be useful in measuring DNA synthesis with SPECT. PMID- 14530486 TI - Detection of gastric and colonic sentinel nodes through endoscopic administration of 99mTc-DTPA-mannosyl-dextran in pigs. AB - The purpose of this study was to develop a method for the endoscopic administration of a radiopharmaceutical for sentinel node detection and to characterize its uptake by gastric and colonic sentinel nodes. (99m)Tc Diethylenetriaminepentaacetic acid (DTPA)-mannosyl-dextran is a new radiotracer labeled with (99m)Tc and composed of multiple units of mannose attached to the polymeric backbone, dextran. METHODS: Gastric and colonic lymph node detection was studied in 4 fasting and anesthetized pigs. A flexible video endoscope was inserted into the rectum and positioned in the lower colon or advanced down the esophagus into the stomach. A standard endoscopic sclerotherapy needle and sheath filled with 0.9% saline was then loaded with 0.2 mL of a 7.4-MBq 1:1 (v/v) mixture of isosulfan blue and (99m)Tc-DTPA-mannosyl-dextran (7.4 MBq, 0.3 nmol). The needle and sheath were passed through the biopsy channel, and the radiotracer/dye solution (0.1 mL) was injected in a tangential fashion into the submucosa. Within 5-10 min after injection, all radioactive or blue lymph nodes were excised, assayed for radioactivity, and noted for color. Distal lymph nodes were also excised, assayed, and noted for color. RESULTS: (99m)Tc-DTPA-mannosyl dextran uptake by colonic sentinel nodes (n = 4) ranged from 0.54% to 2.4% of the injected dose; all radioactive nodes were stained blue. The uptake for all (n = 3) distal nodes ranged from 0.001% to 0.005%, and none of these nodes were stained blue. Uptake by gastric sentinel nodes (n = 6) ranged from 0.13% to 4.50%; all radioactive nodes were stained blue. The range for distal nodes was 0.001% to 0.050%; no distal nodes were stained blue. In 2 pigs, each gastric injection produced 2 hot and blue lymph nodes. CONCLUSION: Within 10 min of a gastric and colonic submucosal injection, (99m)Tc-DTPA-mannosyl-dextran demonstrated high sentinel node uptake and high concordance with isosulfan blue. PMID- 14530487 TI - Evaluation of 18F-CPFPX, a novel adenosine A1 receptor ligand: in vitro autoradiography and high-resolution small animal PET. AB - Adenosine modulates brain activity through 4 G protein-coupled receptors, primarily adenosine A(1) receptors (A(1)ARs). A(1)ARs are heterogeneously distributed throughout the brain and participate in many physiologic processes for example, the induction of sleep and feedback inhibition of excitatory neurotransmission. There is also evidence that A(1)ARs are involved in brain pathologies, including cerebral ischemia, epilepsy, and neurodegeneration. Therefore, measuring A(1)ARs in the living brain has been a long-standing goal. This report describes the preclinical evaluation of (18)F-8-cyclopentyl-3-(3 fluoropropyl)-1-propylxanthine ((18)F-CPFPX), a novel A(1)AR PET ligand. METHODS: CPFPX, a xanthine-based A(1)AR antagonist, was labeled with either (18)F or (3)H, maintaining identical chemical structures, and evaluated in rats as a putative radioligand for in vivo or in vitro imaging of brain A(1)ARs by quantitative receptor autoradiography and the combination of high-resolution small animal PET and MRI. RESULTS: (3)H-CPFPX bound with nanomolar affinity (K(d), 4.4 nmol/L) to A(1)ARs and showed a distribution typical of cerebral A(1)ARs. In extensive in vitro competition studies, (3)H-CPFPX proved to be a highly selective and specific A(1)AR radioligand. Neither the nonxanthine-type adenosine A(2A) receptor antagonist ZM 241385 nor multiple cholinergic, serotoninergic, and glutamatergic receptor compounds competed for (3)H-CPFPX below the micromolar level. In vivo animal PET and ex vivo autoradiographic experiments measured radioactivity in discrete brain regions after intravenous injection of (18)F CPFPX. (18)F-CPFPX had excellent in vivo stability and penetrated the blood-brain barrier immediately after injection due to its high lipophilicity. Brain uptake was rapid and particularly high in gray matter regions. Retention of (18)F-CPFPX was highest in the cerebellum, thalamus, and neocortex with evidence of saturable binding. Low binding potentials were found in the midbrain. In vivo displacement PET experiments with the A(1)AR antagonist 8-cyclopentyl-1,3-dipropylxanthine showed a 72% +/- 8% displacement of (18)F-CPFPX. CONCLUSION: (18)F-CPFPX is a highly selective and specific ligand for A(1)ARs and a suitable radioligand for noninvasive PET imaging of A(1)ARs in the living brain. These studies also support the application of high-resolution animal PET as an effective in vivo imaging tool in the evaluation process of new radioligands. PMID- 14530488 TI - Dosimetric analysis of radioimmunotherapy with 186Re-labeled bivatuzumab in patients with head and neck cancer. AB - From December 1999 until July 2001, a phase I dose escalation study was performed with (186)Re-labeled bivatuzumab, a humanized monoclonal antibody against CD44v6, on patients with inoperable recurrent or metastatic head and neck cancer. The aim of the trial was to assess the safety and tolerability of intravenously administered (186)Re-bivatuzumab and to determine the maximum tolerated dose (MTD) of (186)Re-bivatuzumab. The data were also used for dosimetric analysis of the treated patients. Dosimetry is used to estimate the absorbed doses by nontarget organs, as well as by tumors. It can also help to explain toxicity that is observed and to predict organs at risk because of the therapy given. METHODS: Whole-body scintigraphy was used to draw regions around sites or organs of interest. Residence times in these organs and sites were calculated and entered into the MIRDOSE3 program, to obtain absorbed doses in all target organs except for red marrow. The red marrow dose was calculated using a blood-derived method. Twenty-one studies on 18 patients, 5 female and 16 male, were used for dosimetry. RESULTS: The mean red marrow doses were 0.49 +/- 0.03 mGy/MBq for men and 0.64 +/ 0.03 mGy/MBq for women. The normal organ with the highest absorbed dose appeared to be the kidney (mean dose, 1.61 +/- 0.75 mGy/MBq in men and 2.15 +/- 0.95 mGy/MBq in women; maximum kidney dose in all patients, 11 Gy), but the doses absorbed are not expected to lead to renal toxicity. Other organs with doses exceeding 0.5 mGy/MBq were the lungs, the spleen, the heart, the liver, the bones, and the testes. The doses delivered to the tumor, recalculated to the MTD level of 1.85 GBq/m(2), ranged from 3.8 to 76.4 Gy, with a median of 12.4 Gy. A good correlation was found between platelet and white blood cell counts and the administered amount of activity per kilogram of body weight (r = -0.79). CONCLUSION: Dosimetric analysis of the data revealed that the range of doses to normal organs seems to be well within acceptable and safe limits. Tumor doses ranged from 4 to 76 Gy. Given the acceptable tumor doses, (186)Re-labeled bivatuzumab could be a good candidate for future adjuvant radioimmunotherapy in patients with minimal residual disease. PMID- 14530489 TI - Synthesis and in vivo evaluation of 18F-desbromo-DuP-697 as a PET tracer for cyclooxygenase-2 expression. AB - Cyclooxygenase-2 (COX-2) overexpression has been observed in various pathologies, such as inflammation, cancer, ischemia, and Alzheimer's disease. As an initial step toward a noninvasive PET technique to assay COX-2 expression, this study describes the synthesis and preliminary evaluation of the radiolabeled COX-2 inhibitor (18)F-desbromo-DuP-697. METHODS: Desbromo-DuP-697 was radiolabeled by a nucleophilic aromatic substitution reaction of the nitro precursor with (18)F fluoride. Biodistribution studies of the tracer were performed in a carrageenan induced hyperalgesia rat model. Brain uptake was investigated with autoradiography. To confirm the results of the biodistribution, COX activity was determined by a peroxidase assay. RESULTS: Biodistribution studies showed specific binding of the tracer to COX-2 in heart, kidney, brain, and blood cells, but not in the inflamed paw, which was probably due to low COX-2 expression. In the brain, regional differences in tracer uptake were observed, with high uptake in cortical regions. (18)F-Desbromo-DuP-697 did not show any binding to COX-1. Nonspecific uptake was high in fat and intestines. CONCLUSION: Because of its ability to cross the blood-brain barrier, (18)F-desbromo-DuP-697 appears to be suitable for COX-2 imaging in the brain. Its high nonspecific uptake in the intestines may limit its use for imaging in the abdominal region. PMID- 14530490 TI - Mean transit time and functional image in asialoglycoprotein receptor scintigraphy. PMID- 14530491 TI - Underestimation of absorbed dose to kidney. PMID- 14530492 TI - Overlooked contributions on sodium iodide symporter. PMID- 14530493 TI - Diagnosis and management of central nervous system metastases from breast cancer. AB - The brain, cranial nerves, leptomeninges, spinal cord, and eye compose the central nervous system (CNS) and are at risk for the development of metastases from breast cancer. Such metastases are diagnosed on the basis of clinical suspicion and substantiated by neuroimaging, resection when indicated, and sampling of cerebrospinal fluid when leptomeningeal metastasis (LM) is suspected. Treatment is aimed at palliation of symptoms and preservation of neurologic function. Historically, conventional radiation therapy has been the mainstay of palliative treatment for brain, cranial nerve, spinal cord, and ocular metastases. However, additional treatment options for brain metastases have been brought about by technological advances in surgery to resect brain metastases, and stereotactic radiosurgery (SRS) to focally irradiate metastases, both of which have been substantiated by data from randomized trials. Ongoing research is aimed at refining criteria to select which patients with brain metastases should undergo surgery and SRS and how these focal therapies should be optimally integrated with whole-brain radiotherapy. Therapy for LM must carefully balance the potential risks and perceived benefits associated with CNS-directed therapies. Despite advances in neuroimaging, surgery, and radiation therapy, novel treatments are needed to improve the effectiveness of treatments for CNS metastases, especially LM, while reducing attendant neurotoxicity. PMID- 14530494 TI - The role of ABC transporters in clinical practice. AB - Drug resistance remains one of the primary causes of suboptimal outcomes in cancer therapy. ATP-binding cassette (ABC) transporters are a family of transporter proteins that contribute to drug resistance via ATP-dependent drug efflux pumps. P-glycoprotein (P-gp), encoded by the MDR1 gene, is an ABC transporter normally involved in the excretion of toxins from cells. It also confers resistance to certain chemotherapeutic agents. P-gp is overexpressed at baseline in chemotherapy-resistant tumors, such as colon and kidney cancers, and is upregulated after disease progression following chemotherapy in malignancies such as leukemia and breast cancer. Other transporter proteins mediating drug resistance include those in the multidrug-resistance-associated protein (MRP) family, notably MRP1, and ABCG2. These transporters are also involved in normal physiologic functions. The expressions of MRP family members and ABCG2 have not been well worked out in cancer. Increased drug accumulation and drug resistance reversal with P-gp inhibitors have been well documented in vitro, but only suggested in clinical trials. Limitations in the design of early resistance reversal trials contributed to disappointing results. Despite this, three randomized trials have shown statistically significant benefits with the use of a P-gp inhibitor in combination with chemotherapy. Improved diagnostic techniques aimed at the selection of patients with tumors that express P-gp should result in more successful outcomes. Further optimism is warranted with the advent of potent, nontoxic inhibitors and new treatment strategies, including the combination of new targeted therapies with therapies aimed at the prevention of drug resistance. PMID- 14530495 TI - Chemoembolization in the management of liver tumors. AB - The dual vascular supply of the liver affords a unique opportunity to explore intraarterial therapies for hepatic malignancies. Chemoembolization is a well established technique combining intra-arterial chemotherapy with delivery of embolic agents in order to achieve an antitumor effect due to a high local concentration and prolonged dwell time of the drug, along with select ischemia. Many tumors, such as hepatocellular carcinoma, colorectal cancer, and neuroendocrine tumors, cause symptoms and death by local growth and destruction of the liver. While there are other methods capable of controlling small or isolated hepatic neoplasms, none are suitable for the majority of these patients. Chemoembolization can produce significant results in terms of tumor shrinkage in many of these patients, and there are studies to suggest a survival advantage in hepatocellular carcinoma. Toxicity, however, may be substantial, and patient selection is crucial in order to achieve the optimal benefit of this powerful technique for individual populations. PMID- 14530496 TI - PET scans in the staging of lymphoma: current status. AB - Positron emission tomography (PET) is a novel functional imaging technique that provides several inherent advantages over conventional nuclear scintigraphy. Several studies have suggested a role for PET using the positron emitter fluorine 18 in the diagnosis and follow-up of patients with lymphoma. This review summarizes the existing data evaluating the role of 2-fluoro-2-deoxy-D-glucose (FDG)-PET in both the staging and follow-up of patients with lymphoma. Most studies of PET involve patients with either Hodgkin's disease or diffuse large B cell non-Hodgkin's lymphoma. PET detects more disease sites above and below the diaphragm on staging of lymphoma than gallium scintigraphy and may have particular utility in the evaluation of the spleen. Moreover, persistently positive PET scans during and after chemotherapy appear to have a high sensitivity for predicting subsequent relapse. A negative PET scan at the end of therapy provides very favorable prognostic information. Persistently positive PET scans at the end of therapy warrant close follow-up or additional diagnostic procedures, since some of those patients may remain in prolonged remission. Clearly, additional studies, including prospective blinded trials and cost effectiveness analyses, are warranted to determine which subsets of patients with lymphoma ultimately will benefit from this modality. PMID- 14530497 TI - Management of human melanoma: what has the last decade wrought? PMID- 14530498 TI - Pros and cons of adjuvant interferon in the treatment of melanoma. AB - Should interferon alpha (IFN-alpha) be considered the standard of care for the adjuvant therapy of high-risk malignant melanoma? For 2003, it was estimated that 51,400 cases of invasive melanoma would be diagnosed. The risk of recurrence after surgery is reported to be approximately 60% for patients with thick primary lesions (T4N0M0, American Joint Committee on Cancer [AJCC] stage IIB) and 75% for patients with regional nodal metastases (T1-4N1M0, AJCC stage III). The observation that melanoma is susceptible to attack by the host's immune system has resulted in the testing of a remarkably broad spectrum of immunotherapies in the adjuvant setting. Many of these approaches failed to demonstrate a significant clinical impact, until the use of adjuvant IFN-alpha. Conflicting data from several large, randomized clinical trials resulted in a rapid rise and then decline in the use of IFN-alpha in the adjuvant setting. This roller coaster has left many clinicians still hesitant to strongly recommend it, and the use of adjuvant IFN-alpha in high-risk melanoma remains controversial. This manuscript reviews the leading arguments for and against its routine use and addresses questions regarding its role in the management of high-risk malignant melanoma. PMID- 14530500 TI - Genetics and the management of women at high risk for breast cancer. AB - It is estimated that 5%-10% of all breast cancers in women are associated with hereditary susceptibility due to mutations in autosomal dominant genes, such as BRCA1 and BRCA2, p53, pTEN, and STK11/LKB1. Another 15%-20% of female breast cancers occur in women with a family history but without an apparent autosomal dominant inheritance pattern, and are probably due to other genetic factors with environmental influence. Approximately 7%-10% of ovarian cancers occur in women with hereditary susceptibility, primarily secondary to mutations in BRCA1 and BRCA2, with smaller contributions from mutations in mismatch repair genes associated with the hereditary nonpolyposis colorectal cancer and other, as yet undiscovered, genes. PMID- 14530499 TI - The melanoma epidemic: res ipsa loquitur. AB - Many have debated whether or not we are in the midst of a melanoma epidemic. Some facts are clear and helpful to this debate, while others are less clear. The incidence and mortality of melanoma have increased over the last several decades, but the incidence has risen faster than the mortality. The incidence has risen 3% 7% on average over several decades and even more rapidly among Caucasian men and the elderly. In the U.S., the incidence in men is higher than in women after the age of 40, and the difference between men and women increases from age 40 until the end of life. The incidence in the U.S. has risen most rapidly among in situ and localized lesions, but distant and regional disease have increased as well. Among localized disease, in the U.S. from 1988-1997, all stages increased by comparable amounts. This strongly argues against the idea that the increase in incidence of melanoma is only due to early detection of thin lesions or biologically benign lesions, at least during the time period studied. On the other hand, early detection of thin lesions may well account for lower increases in mortality than incidence and improvements in survival. Survival has increased from approximately 60% in the 1960s to 89% in recent years. Improvements in survival appear to be related to earlier diagnosis, rather than an improvement in survival of a given stage. Studies consistently point to a major role for UV light exposure as the most important risk factor for those individuals with a phenotypic susceptibility. Public health efforts aim at primary and secondary prevention strategies. Primary prevention strategies attempt to prevent people from developing melanoma, primarily through avoiding exposure to UV light. There is a particular emphasis on avoidance of UV light exposure in childhood and young adulthood, when it appears the risk is greatest. When strict avoidance cannot be adhered to, sunscreens have been logically recommended. Secondary prevention strategies include screening campaigns and educational campaigns. Many of these strategies appear promising but require further rigorous testing. The melanoma epidemic has arisen for a variety of reasons including: a true increase in melanomas of malignant behavior, a particularly high increase in localized and in situ lesions, and an increase in the number of biopsies performed, which may have resulted in an increased detection of less aggressive lesions. The contribution of possible changes in the diagnostic criteria for melanoma to the increased incidence remains unknown. PMID- 14530501 TI - Andropause: symptom management for prostate cancer patients treated with hormonal ablation. AB - Andropause, or the age-related decline in serum testosterone, has become a popular topic in the medical literature over the past several years. Andropause includes a constellation of symptoms related to lack of androgens, including diminished libido, decreased generalized feeling of well-being, osteoporosis, and a host of other symptoms. The andropause syndrome is very prominent in men undergoing hormonal ablation therapy for prostate cancer. Most significant in this population are the side effects of hot flashes, anemia, gynecomastia, depression, cognitive decline, sarcopenia, a decreased overall quality of life, sexual dysfunction, and osteoporosis with subsequent bone fractures. The concept of andropause in prostate cancer patients is poorly represented in the literature. In this article, we review the current literature on the symptoms, signs, and possible therapies available to men who cannot take replacement testosterone. PMID- 14530502 TI - The Kenneth B. Schwartz Center at Massachusetts General Hospital hematology oncology department: hope for the homeless. 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 nonprofit organization dedicated to supporting and advancing compassionate health care delivery, which provides hope to the patient and support to caregivers and encourages the healing process. The Center sponsors the Schwartz Center Rounds, a monthly multidisciplinary forum during which caregivers discuss a specific cancer patient, reflect on the important psychological issues faced by patients, their families, and their caregivers, and gain insight and support from their fellow staff members. A homeless man with head and neck cancer presents to the emergency room: a sad and familiar story. But this story is redeemed by his 35-year friendship with a priest, a man whose unconditional love and support became critical to the patient's care and treatment. The patient had lived for 30 years in homeless shelters, had problems with alcohol abuse, and was notoriously noncompliant with medical caregivers. He could not speak due to his disease, was illiterate with limited intellectual capacity, and had neither a job nor a family. Despite huge and apparently insurmountable problems for the patient, the oncology team was able to carve out a package of care, successfully communicate, and mobilize a support network to allow successful completion of chemoradiation therapy. The team developed a strong commitment to his care and an affectionate bond, which very positively affected all of those involved. We discuss issues of access to cancer care, and the special problems presented by homeless patients. PMID- 14530503 TI - The molecular perspective: epidermal growth factor. PMID- 14530505 TI - Questions about thimerosal remain. PMID- 14530507 TI - Apoptosis, chemoresistance, and breast cancer: insights from the MCF-7 cell model system. AB - The MCF-7 cell line was derived from a patient with metastatic breast cancer in 1970. Since then it has become a prominent model system for the study of estrogen receptor-positive breast cancer. With this model as a focus, this review summarizes important studies addressing tumor necrosis factor-alpha as a prototypical apoptosis-inducing cytokine in MCF-7 cells. Both survival and death receptor signaling pathways are discussed in terms of their role in chemotherapy induced apoptosis as well as in chemoresistance. Novel therapeutic approaches to the treatment of breast cancer are proposed utilizing knowledge of these signaling pathways as targets. Specifically, ceramide metabolism is proposed as a novel target for chemosensitivity, perhaps combined with selective inhibitors of Bcl-2 or PI3K/Akt/nuclear factor-kappaB. Suggested areas of future research include translational studies manipulating candidate survival and death signaling pathways. PMID- 14530508 TI - Genetics of pituitary tumors: Focus on G-protein mutations. AB - In recent years the demonstration that human pituitary adenomas are monoclonal in origin has provided further evidence that pituitary neoplasia arise from the replication of a single mutated cell in which growth advantage results from either activation of proto-oncogenes or inactivation of tumor suppressor genes. While common oncogenes, such as Ras, are only exceptionally involved, the only mutations identified in a significant proportion of pituitary tumors, and particular in GH-secreting adenomas, occur in the Gsalpha gene (GNAS1) and cause constitutive activation of the cAMP pathway (gsp oncogene). Moreover, pituitary tumors overexpress hypothalamic releasing hormones, growth factors, and their receptors as well as cyclins involved in cell cycle progression. As far as the role of tumor suppressor genes in pituitary tumorigenesis is concerned, reduced expression of these genes seems to frequently occur in pituitary tumors as a consequence of abnormal methylation processes. Although the only mutational change so far identified in pituitary tumors is the gsp oncogene, this oncogene is not associated with a clear phenotype in patients bearing positive tumors. Mechanisms able to counteract the cAMP pathway, such as high sensitivity to somatostatin, and induction of genes with opposite actions, such as phosphodiesterases, CREB end ICER, or instability of mutant Gsalpha, have been proposed to account for the lack of genotype/phenotype relationships. PMID- 14530509 TI - Extracellular matrix degradation by cultured mesangial cells: mediators and modulators. AB - Decreased degradation of the glomerular extracellular matrix (ECM) is thought to contribute to the accumulation of glomerular ECM that occurs in diabetic nephropathy and other chronic renal diseases. Several lines of evidence indicate a key role for the plasminogen activator/plasminogen/plasmin system in glomerular ECM degradation. However, which of the two plasminogen activators (PAs) present in renal tissue, tissue plasminogen activator (tPA) or urokinase-type plasminogen activator (uPA), is responsible for plasmin generation and those factors that modulate the activity of this system remain unclear. This study utilized mesangial cells isolated from mice with gene deletions for tPA, uPA, and plasminogen activator inhibitor 1 (PAI-1) to further delineate the role of the PA/plasminogen/plasmin system in ECM accumulation. ECM degradation by uPA-null mesangial cells was not significantly different from controls (92% +/- 1%, n = 12). In contrast, ECM degradation by tPA-null mesangial cells was markedly reduced (-78 +/- 1%, n = 12, P < 0.05) compared with controls, whereas tPA/uPA double-null mesangial cells degraded virtually no ECM. Previous studies from this laboratory have established that transforming growth factor-beta1 (TGFbeta1) inhibits ECM degradation by cultured mesangial cells by increasing the production of PAI-1, the major physiological PA inhibitor. In keeping with this observation, TGFbeta1 (1 ng/ml) had no effect on ECM degradation by PAI-1-null MC. High glucose levels (30 mM) in the presence or absence of insulin (0.1 mM) caused a moderate increase in ECM degradation by normal human mesangial cells. In contrast, glycated albumin, whose concentration is known to increase in diabetes, produced a dose-dependent (0.2-0.5 mg/ml) inhibition of ECM degradation by normal human mesangial cells. Taken together, these results document the importance of tPA versus uPA in renal plasmin production and indicate that in contrast to elevated glucose, glycated albumin may contribute to ECM accumulation in diabetic nephropathy. PMID- 14530510 TI - Negative regulation of interleukin-12 production by a rapamycin-sensitive signaling pathway: a brief communication. AB - Interleukin-12 (IL-12), an important cytokine in host defense against microbial pathogens, regulates natural killer and T-cell function(s) including the induction of gamma-interferon production. The major cellular sources of IL-12 are monocytes/macrophages. Bacteria, bacterial products, and intracellular parasites are the most efficient inducers of IL-12 production. In the present study we show that a signal transduction pathway sensitive to rapamycin may have an important role in the regulation/suppression of Staphylococcus aureus-induced IL-12 production in vitro. Human peripheral blood mononuclear cells, monocytes, or a human monocytic cell line THP-1 were stimulated with S. aureus Cowan strain 1 (SAC) in the presence or absence of rapamycin and investigated for production of IL-12 protein by enzyme-linked immunosorbent assay and IL-12 p40 mRNA accumulation by RNase protection assay or real-time quantitative polymerase chain reaction. The results show that rapamycin significantly enhances SAC-induced IL 12 p70 protein production and IL-12 p40 mRNA accumulation. Further the results demonstrate that wortmannin enhances SAC-induced IL-12 p40 mRNA accumulation, whereas Ly294002 does not. These data indicate that a rapamycin-sensitive signaling pathway may act as a negative feedback cascade in the regulatory mechanisms of IL-12 production. PMID- 14530511 TI - Evidence that growth hormone exerts a feedback effect on stomach ghrelin production and secretion. AB - Ghrelin is a recently discovered stomach hormone that stimulates pituitary growth hormone (GH) secretion potently. The purpose of these experiments was to test the hypothesis that a stomach-ghrelin-pituitary-GH axis exists in which either an elevation or reduction in systemic GH levels will exert a negative or positive feedback action, respectively, on stomach ghrelin homeostasis. In rats, GH administration decreased stomach ghrelin mRNA levels and plasma ghrelin levels significantly. In GH-releasing hormone (GHRH) transgenic mice, GHRH overexpression decreased stomach ghrelin peptide levels when compared with control mice. In aged rats (25 months) stomach ghrelin mRNA and peptide levels and plasma ghrelin levels were decreased when compared with young rats (5 months). Because GH secretion is reduced in aged rats, the elevated stomach ghrelin production and secretion may reflect a decreased GH feedback on stomach ghrelin, homeostasis, and secretion. Together, these findings suggest that endogenous pituitary GH exerts a feedback action on stomach ghrelin homeostasis and support the hypothesis that a stomach-ghrelin-pituitary GH axis exists. PMID- 14530512 TI - Erythropoietin-induced metallothionein gene expression: role in proliferation of K562 cells. AB - Recent evidence has demonstrated an appreciable expression of metallothionein (MT) in erythrocytes. However, the induction of the MT protein by hematopoietic growth factors and its subsequent functional significance on clonal expansion or differentiation of erythroid progenitor cells remain elusive. We therefore examined the effects of growth factors erythropoietin (EPO), granulocyte-monocyte colony-stimulating factor (GM-CSF), and interleukin-3 (IL-3) on MT gene expression in erythroid progenitor K562 cell line. EPO, but not IL-3 or GM-CSF, induced a 3-fold increase in MT transcripts in K562 cells. MT induction was associated with EPO-induced cellular proliferation, suggesting a specific role for MT induction by EPO in erythroid progenitor cells. However, EPO- or sodium butyrate-induced differentiation as monitored by hemoglobin formation was inhibited in K562 cells stably transfected with an expression vector containing human MT-IIA gene. This inhibition of differentiation was partially reversed in these cells by an MT antisense phosphorothioate oligonucleotide. Furthermore, the MT-induced inhibition of differentiation was associated with downregulation of EPO receptor transcripts in K562 cells. These data suggest that, among growth factors required for erythropoiesis, EPO is a potent inducer of MT, and that MT may plays a significant role in EPO-induced proliferation, but not in the erythroid-specific differentiation of the progenitor cells. PMID- 14530513 TI - Diabetes (db/db) mutation-induced ovarian involution: progressive hypercytolipidemia. AB - Ovarian atrophy and reproductive tract incompetence are recognized consequences of the progressive expression of the overt, diabetes-obesity syndrome (DOS) in C57BL/KsJ (db/db) mutant mice. The present studies evaluated the progressive changes in ovarian cytoarchitecture, endocrine expression, and reproductive tract cytolipidemic parameters that promote reproductive failure and ovarian involution during the pre-onset, initial, progressive, and chronic expression stages of the DOS. Paired littermate control (normal: +/?) and diabetic (mutant: db/db) C57BL/KsJ females were selected for analysis of ovarian parameters at 2 weeks (pre-onset expression of DOS), 4 weeks (initial DOS expression), 8 weeks (progressive DOS: hyper-glycemic/lipidemic), and 16 weeks (overt/chronic DOS expression) of age. All 4- to 16-week-old (db/db) groups were obese, hyperglycemic, and hyperinsulinemic as compared with age-matched (+/?) controls. Prior to phenotypic expression of the DOS (2 week groups), ovarian interstitial cytolipidemia characterized the perifollicular and cortical regions of db/db tissue samples relative to +/? indices, while comparable body weight, blood glucose, as well as serum insulin and ovarian steroid hormone concentrations characterized both the +/? and db/db groups. Overt DOS expression in the 4-week old db/db groups was characterized by body obesity, systemic hyperglycemia hyperinsulinemia, and extensive hypercytolipidemia of ovarian folliculothecal compartments, as well as enhanced tissue lipase activities. By 8 weeks of age, progressive hypercytolipidemia characterized interstitial, thecal, and follicular granulosa cell layers of db/db tissue samples concurrent with suppressed ovarian steroid hormone production, enhanced lipid sequestration, and exacerbation of systemic hyper-glycemia/insulinemia. By 16 weeks of age, the chronic-DOS was characterized by extensive ovarian follicular involution, cortical perivascular hyperlipidemic infiltration, thecal cell atrophy, and follicular granulosa lipid imbibition. These data indicate that db/db mutation-induced ovarian structural and functional involution is a direct reflection of the cellular metabolic shift towards lipogenesis, indicated by the progressive cytoarchitectural transformation into adipocyte-like entities. The cytological indications of cellular metabolic compromise, which precede the phenotypic expression of the DOS indices, suggests that correction of these abnormal shifts in ovarian endocrine and cellular metabolism may restore, delay, or prevent the further compromise of ovarian function by db/db mutation expression. PMID- 14530514 TI - Regulation of human immune gene expression as influenced by a commercial blended Echinacea product: preliminary studies. AB - Consumption of Echinacea at the first sign of symptoms has been clinically shown to reduce both the severity and duration of cold and flu. Quantitative polymerase chain reaction optimized for precision and reproducibility was utilized to explore in vitro and in vivo changes in the expression of immunomodulatory genes in response to Echinacea. In vitro exposure of THP-1 cells to 250 microg/ml of Echinacea species extracts induced expression (up to 10-fold) of the interleukin 1alpha, interleukin-1beta, tumor necrosis factor-alpha, intracellular adhesion molecule, interleukin-8, and interleukin-10 genes. This preliminary result is consistent with a general immune response and activation of the nonspecific immune response cytokines. In vivo gene expression within peripheral leukocytes was evaluated in six healthy nonsmoking subjects (18-65 years of age). Blood samples were obtained at baseline and on Days 2, 3, 5, and 12 after consuming a commercial blended Echinacea product, three tablets three times daily (1518 mg/day) for two days plus one additional dose (506 mg) on day three. Serum chemistry and hematological values were not different from baseline, suggesting that liver or bone marrow responses were not involved in acute responses to Echinacea. The overall gene expression pattern at 48 hr to 12 days after taking Echinacea was consistent with an antiinflammatory response. The expression of interleukin-1beta, tumor necrosis factor-alpha, intracellular adhesion molecule, and interleukin-8 was modestly decreased up through Day 5, returning to baseline by day 12. The expression of interferon-alpha steadily rose through Day 12, consistent with an antiviral response. These preliminary data present a gene expression response pattern that is consistent with Echinacea's reported ability to reduce both the duration and intensity of cold and flu symptoms. PMID- 14530515 TI - Nitric oxide as a local mediator of prostaglandin F2alpha-induced regression in bovine corpus luteum: an in vivo study. AB - To test whether nitric oxide (NO) is involved in prostaglandin (PG) F2alpha induced regression of the bovine corpus luteum (CL) in vivo, heifers were treated as follows: Group 1, saline (3 ml/h); Group 2, dinoprost, an analogue of prostaglandin F2alpha (aPGF2alpha; 5 mg/0.5 h); Group III, Nomega-nitro-L arginine methyl ester (L-NAME; 200 mg/4 h), an inhibitor of nitric oxide synthase; and Group IV, L-NAME (400 mg/4 h) and aPGF2alpha (5 mg/0.5 h). All treatments were administered by an intraluteal microdialysis system (MDS) on day 15 of the cycle. Perfusate and jugular plasma samples were collected at half-hour intervals; additionally, jugular plasma samples were collected once daily from day 16 to day 21 of the cycle. In the perfusate samples, aPGF2alpha increased P4 (P < 0.05), PGE2 (P < 0.001), and LTC4 (P < 0.05) concentrations; L-NAME increased P4 (P < 0.05) but did not change PGE2 and LTC4 (P > 0.05) concentrations as compared with the period before treatment. Simultaneous perfusion of CL with L-NAME and aPGF2alpha caused a further increase of P4 concentration (P < 0.05) induced by L-NAME or aPGF2alpha treatment and increased PGE2 and LTC4 (P < 0.001) concentrations to the level observed after aPGF2alpha treatment. Perfusion of CL with aPGF2alpha caused luteal regression within 24 h, while perfusion with L-NAME prolonged the life span of CL to day 21 (P < 0.05). Concomitant L-NAME and aPGF2alpha treatment partially counteracted (P < 0.05) the luteal regression caused by aPGF2alpha administration. These results show that NO is involved in the process of luteolysis in the bovine CL and suggest that the luteolytic effect of aPGF2alpha may be mediated by NO as an important component of an autocrine/paracrine luteolytic cascade. PMID- 14530516 TI - Soy protein with or without isoflavones, soy germ and soy germ extract, and daidzein lessen plasma cholesterol levels in golden Syrian hamsters. AB - Dietary isolated soy protein (ISP, containing approximately equal amounts of daidzein and genistein), ethanol-extracted ISP (ISP (-)), soygerm or soygerm extract (containing large amounts of daidzein and glycitein and little genistein) and the isoflavone, daidzein, were hypothesized to lessen plasma cholesterol in comparison with casein. Sixty male and 60 female golden Syrian hamsters (6-8 weeks of age) were randomly assigned to six treatments fed for 10 weeks. Four of the experimental diets (ISP, daidzein, soygerm, and soygerm extract) contained 1.3 mmol total isoflavones/kg. The ISP (-) diet contained 0.013 mmol isoflavone/kg, whereas the casein diet contained no isoflavones. Hamsters fed ISP, ISP (-), daidzein, soygerm, and soygerm extract had significantly less plasma total cholesterol (by 16%-28%), less non-HDL cholesterol (by 15%-50%) and less non-HDL/HDL cholesterol ratios compared with hamsters fed casein (P < 0.01). For male hamsters, there were no differences among treatments in plasma HDL concentrations. Female hamsters fed ISP (-) had significantly greater HDL levels (P < 0.01) than females fed casein or daidzein. Triglyceride concentration was significantly less in hamsters fed ISP (-) compared with the casein-fed females. Because soy protein with or without isoflavones, soygerm and soygerm extract, and daidzein lessened plasma cholesterol to an approximately equal extent, soy protein alone, varying mixtures of isoflavones, and other extractable components of soy are responsible for cholesterol-lessening effects of soy foods, mainly due to their effects to lessen LDL cholesterol. PMID- 14530517 TI - Polyamine concentrations in the brain of vitamin B12-deficient rats. AB - To study the pathophysiology of the neuronal degeneration in vitamin B12 deficiency, we investigated the concentrations of the polyamines putrescine, spermidine, and spermine in brain regions and liver using high-performance liquid chromatography with fluorescence detection. Male Wistar rats were fed either a control or vitamin B12-deficient diet for 20 weeks. No remarkable behavioral changes were observed. Serum vitamin B12 and hepatic methionine concentrations were significantly lower and hepatic homocysteine was elevated in rats fed vitamin B12-deficient diet than in controls. Vitamin B12 deficiency was associated with decreased concentrations of spermidine, spermidine in liver and some regions of brain, although there were no observed abnormalities in behavior. These results suggest that vitamin B12 deficiency may play a role in neuronal degeneration through the disturbance of polyamine concentrations in rat brain. PMID- 14530518 TI - Influence of maternal stress on uranium-induced developmental toxicity in rats. AB - It has been demonstrated that uranium is an embryo/fetal toxicant when given orally or subcutaneously to pregnant mice. On the other hand, maternal stress has been shown to enhance the developmental toxicity of a number of metals. In this study, maternal toxicity and developmental effects of a concurrent exposure to uranyl acetate dihydrate (UAD) and restraint stress were evaluated in rats. Four groups of pregnant animals were given subcutaneous injections of UAD at 0.415 and 0.830 mg/kg/day on Days 6 to 15 of gestation. Animals in two of these groups were also subjected to restraint for 2 hr/day during the same gestational days. Control groups included restrained and unrestrained pregnant rats not exposed to UAD. Cesarean sections were performed on gestation Day 20, and the fetuses were weighed and examined for malformations and variations. Maternal toxicity and embryotoxicity were noted at 0.830 mg/kg/day of UAD, while fetotoxicity was evidenced at 0.415 and 0.830 mg/kg/day of UAD by significant reductions in fetal body weight and increases in the total number of skeletally affected fetuses. No teratogenic effects were noted in any group. Maternal restraint enhanced uranium induced embryo/fetal toxicity only at 0.830 mg/kg/day, a dose that was also significantly toxic to the dams. As in previous studies with other metals, maternal stress enhances uranium-induced developmental toxicity at uranium doses that are highly toxic to the dams; however, at doses that are less acutely toxic the role of maternal stress would not be significant. PMID- 14530519 TI - Pendrin transporter carries out iodide uptake into MCF-7 human mammary cancer cells. AB - Previous studies have shown that iodide is actively taken up into mammary alveolar epithelial cells and secreted into milk. In the present studies we demonstrate that 125I also accumulates in MCF-7 cells against a concentration gradient; distribution ratios of greater than 30 were achieved. Iodide uptake into MCF-7 cells is transient, with peak accumulations occurring in about 5 min. The iodide is rapidly metabolized, probably to iodine, and it then exits the cells. The iodide transporter identified in MCF-7 cells is pendrin. DIDS, a nonspecific inhibitor of anion exchange, inhibits iodide uptake. Iodide uptake is impaired at reduced temperature, but is not dependent on sodium. Inhibitors of the sodium-iodide symporter (NIS) as well as ouabain did not affect the extent of iodide uptake. The pendrin transporter but not NIS was identified via western blotting techniques. Pendrin appears to be the primary iodide transporter in the MCF-7 cell line stocks that were employed for these studies. PMID- 14530520 TI - Thyrotropin secretagogues reduce rat pituitary neuromedin B, a local thyrotropin release inhibitor. AB - Neuromedin B (NB), a bombesin-like peptide, highly concentrated in rat pituitary gland, has been shown to act as an autocrine/paracrine inhibitor of thyrotropin (TSH) release. Here it is shown that a single injection of thyrotropin-releasing hormone (TRH, 1.5 microg/animal, ip), the most important stimulator of thyrotropin secretion, induced approximately 35%-45% decrease in pituitary NB content in rats, as well as an important decrease in NB mRNA at 15 and 30 min (P < 0.05). Acute cold exposure, which induced higher serum TSH with a peak at 30 min, was associated with progressive decrease in pituitary NB, starting at 15 min although only reaching statistical significance after 2 hr (P < 0.05). Although not involved in the early peak, the decrease in NB may be contributing to maintenance of higher serum TSH in cold-exposed animals compared with those at room temperature. Fed rats, 2 hr after being subcutaneously injected with mouse recombinant leptin (8 microg /100 g body wt), showed a x2 increase in serum TSH and 38% reduction in pituitary NB (P < 0.05). In conclusion, TRH and leptin rapidly decreased pituitary NB and it is first proposed that the reduction of the inhibitory tonus of NB on TSH release will ultimately contribute to the amplification of TSH secretion elicited by TSH secretagogues. PMID- 14530521 TI - Molecular mechanism of adaptive cytoprotection induced by ethanol in human gastric cells. AB - Adaptive cytoprotection is the process by which the pretreatment of cells with low concentrations of a noxious agent prevents the damage caused by a subsequent exposure of those cells to higher concentrations of that same agent. In this study, a human gastric carcinoma cell line was used to examine the molecular mechanism of adaptive cytoprotection induced by ethanol. Pretreatment of cells with 1%-4% ethanol made cells resistant to a subsequent exposure to 8% ethanol. This adaptive cytoprotection was accompanied by an increase in prostaglandin E2 synthesis and was partially inhibited by inhibitors of cyclooxygenase-2, but not by an inhibitor of cyclooxygenase-1. Furthermore, the adaptive cytoprotection was not dependent on newly synthesized proteins and was inhibited by a protein tyrosine kinase inhibitor. Based on these results, it is proposed that the stimulation of cyclooxygenase-2-dependent prostaglandin E2 synthesis, which is regulated post-translationally by protein tyrosine phosphorylation, plays an important role in adaptive cytoprotection induced by ethanol in gastric cells. PMID- 14530522 TI - Alloimmune injury and rejection of human skin grafts on human peripheral blood lymphocyte-reconstituted non-obese diabetic severe combined immunodeficient beta2 microglobulin-null mice. AB - Small animal models with the capacity to support engraftment of a functional human immune system are needed to facilitate studies of human alloimmunity. In the present investigation, non-obese diabetic (NOD) severe combined immunodeficient (scid) beta2-microglobulin-null (B2mnull) mice engrafted with human peripheral blood lymphocytes (hu-PBL-NOD-scid B2mnull mice) were used as in vivo models for studying human skin allograft rejection. Hu-PBL-NOD-scid B2mnull mice were established by injection of human spleen cells or PBLs and transplanted with full-thickness allogeneic human skin. Human cell engraftment was enhanced by injection of anti-mouse CD122 antibody. The respective contributions of human CD4+ and CD8+ cells in allograft rejection were determined using depleting antibodies. Human skin grafts on unmanipulated NOD-scid B2mnull mice uniformly survived but on chimeric hu-PBL-NOD-scid B2mnull mice exhibited severe immune mediated injury that often progressed to complete rejection. The alloaggressive hu-PBLs did not require prior in vitro sensitization to elicit targeted effector cell activity. Extensive mononuclear cell infiltration directed towards human origin endothelium was associated with thrombosis and fibrin necrosis. No evidence of graft-versus-host disease was detected. Either CD4+ or CD8+ T cells may mediate injury and alloimmune rejection of human skin grafts on hu-PBL-NOD scid B2mnull mice. It is proposed that Hu-PBL-NOD-scid B2mnull mice engrafted with human skin will provide a useful model for analysis of interventions designed to modulate human allograft rejection. PMID- 14530523 TI - Sudden cardiac death: the war is far from over. PMID- 14530524 TI - Management of febrile convulsion: scene in a regional hospital. AB - OBJECTIVE: To determine whether practice parameters are applied to the management of children with febrile convulsion. DESIGN: Retrospective study. SETTING: Paediatric department of a public hospital, Hong Kong. METHODS: Practice parameters of the American Academy of Pediatrics and audit measures recommended by the Joint Working Group of the Research Unit of the Royal College of Physicians and the British Paediatric Association were employed as standards. Records between January and April 2000 with the diagnostic coding of febrile convulsion, convulsion, status epilepticus, or meningitis/encephalitis/encephalopathy were reviewed. Areas assessed were appropriate documentation of hospital records and unit statistics (adverse outcomes, inappropriate investigations and treatment). RESULTS: Ninety-four consecutive records were evaluated. In the documentation of hospital notes, accurate description of seizure was observed in 92%, incorrect diagnosis or coding in 12%, and presence/absence of signs of meningitis and parental counselling documented in 64% and 85%, respectively. Regarding unit statistics, investigations performed included a complete blood count, blood glucose, serum calcium, serum electrolytes, renal function tests, liver function tests, chest X ray, and urinalysis. The mean number of routine investigations was seven. The average length of stay was 2 days. There were no cases of delay in the diagnosis of central nervous system infection. Inappropriate investigations and treatment were as follows: electroencephalography 11%, computed tomography brain scan 2%, and maintenance anticonvulsants 2%. All patients were discharged home with panadol regardless of clinical state. CONCLUSIONS: The present study showed that the use of unnecessary investigations was common. Investigations, though resulting in significant expense, proved to be of little diagnostic value. Diagnostic procedures should be performed only when specifically called for by the patient's condition or medical history. PMID- 14530526 TI - Large-scale mercury exposure due to a cream cosmetic: community-wide case series. AB - OBJECTIVE: To describe demographic characteristics, patterns of use, reported symptoms, and laboratory results associated with mercury exposure among people who used a beauty cream containing mercury. DESIGN. Descriptive study of a case series. SETTING: The Hong Kong community. PARTICIPANTS: Users of a cream cosmetic who contacted the Department of Health following a public announcement. MAIN OUTCOME MEASURES: Urine and blood mercury concentrations, cream mercury content, self-reported symptoms, duration of cream use, and duration since last cream use. RESULTS: We interviewed 314 cream users, 99% of whom were women. The mean urine and blood mercury concentrations of 286 users, who submitted a urine or blood sample, were 45.2 microg/L (reference level, <20 microg/L) and 17.1 microg/L (reference level, <10 microg/L), respectively; 65% of these participants had elevated mercury concentrations. The mercury content of the cream cosmetic ranged from 660 to 57 000 ppm. Seventy-eight percent of all cream users reported no symptoms, but absence of symptoms was not predictive of low urine and blood mercury concentrations. Urine mercury concentrations were significantly higher among people who last used the cream within 45 days. Blood mercury concentrations were elevated following cream use for as short as 2 days. CONCLUSIONS: The majority of cream users had increased urine or blood mercury concentrations but remained asymptomatic, implying that the incidence of overt symptomatic mercury poisoning resulting from dermal application of creams with mercury content lower than 57 000 ppm is low. Doctors should take a history of the use of cosmetics if patients have clinical or laboratory evidence of mercury exposure; such cases should be reported to public health authorities. PMID- 14530525 TI - Knowledge of cardiopulmonary resuscitation among the public in Hong Kong: telephone questionnaire survey. AB - OBJECTIVES: To evaluate the knowledge of basic life-support and training experience in cardiopulmonary resuscitation among the public in Hong Kong and to identify areas for improvement in public education. DESIGN: Telephone interview using a structured multiple-choice questionnaire. SETTING: Random cross-section of the Hong Kong public, from mid-March to May 2002. PARTICIPANTS: Men and women aged 16 years and older selected using random telephone dialling. MAIN OUTCOME MEASURE: Overall score in the cardiopulmonary resuscitation knowledge questionnaire. RESULTS: Of the 357 participants, approximately 12% had received cardiopulmonary resuscitation training. Cardiopulmonary resuscitation knowledge in Hong Kong was poor, even among the previously trained and especially with regard to circulatory maintenance. The most common reason for not taking cardiopulmonary resuscitation training was lack of time. CONCLUSION: The degree of citizen preparedness in initiating cardiopulmonary resuscitation is very poor in Hong Kong. Intensified educational efforts and exploration of new approaches to improve this first stage in the chain of survival are warranted. PMID- 14530527 TI - Outcome analysis of intraventricular thrombolytic therapy for intraventricular haemorrhage. AB - OBJECTIVES: To evaluate the outcome of intraventricular thrombolytic therapy for intraventricular haemorrhage and to formulate a safe and effective regimen. DESIGN: Retrospective study. SETTING: Regional neurosurgical centre, Hong Kong. PATIENTS: Twenty-nine consecutive adult patients who presented from November 1995 to November 1998 with non-traumatic intraventricular haemorrhage (Graeb score, > or =7) with no active rebleeding risks from vascular abnormalities. INTERVENTIONS: Fourteen consecutive patients received intraventricular streptokinase via the external ventricular drainage, and 15 consecutive patients received intraventricular urokinase treatment. MAIN OUTCOME MEASURES: Patient demographics, Glasgow coma scale score, Graeb score, mortality rate, shunt rate, fever response, infection rate, catheter blockage rate, and local and systemic bleeding tendency. RESULT: The mean age of the 16 men and 13 women was 59 years (range, 14-76 years). The median Graeb score for cases of intraventricular haemorrhage was 10 (range, 7-12). There was no significant difference in terms of the Graeb score distribution, total dosage, and duration of treatment between the streptokinase and urokinase groups. More cases of fever were observed in the streptokinase group, which could be due to its antigenicity. The infection rate of the central nervous system was 3%, and the shunt rate was 24%. The overall 1 month postoperative mortality was 10%, which was related to a low preoperative Glasgow coma scale score (< or =4). No local rebleeding, systemic coagulopathy, or catheter blockage occurred. CONCLUSIONS: Intraventricular thrombolytic therapy is a safe and effective method of managing intraventricular haemorrhage. We suggest instilling 20 000 units urokinase intra-operatively, followed by 20 000 units daily for about 3 days, except in cases of vascular abnormality, bleeding tendency, and trauma. PMID- 14530528 TI - Prognostic factors for successful outcome in patients undergoing controlled ovarian stimulation and intrauterine insemination. AB - OBJECTIVE: To determine the prognostic factors associated with successful outcome following controlled ovarian stimulation and intrauterine insemination. DESIGN: Retrospective analysis. SETTING: University-based assisted reproductive technology centre, Hong Kong. PATIENTS AND METHODS: Patients included 292 couples undergoing 600 treatment cycles, following a standard protocol of human menopausal gonadotrophin injections. Multiple logistic regression analysis was performed to determine which demographic and sperm parameters gave the maximum discrimination to predict pregnancy. RESULTS: One hundred and eleven pregnancies resulted from treatment. The pregnancy rates were 18.5% per cycle and 37.9% per couple. The age of the women was significantly lower for pregnant cycles, and the serum oestradiol levels and number of follicles greater than 16 mm in diameter were significantly higher, compared with non-pregnant cycles. The sperm concentration and number of motile spermatozoa were also significantly increased in pregnant cycles. Pregnancy rate was significantly increased when the raw semen sample contained 20 million/mL or more spermatozoa, normal forms comprised 7% or more, and when the number of motile spermatozoa in inseminated samples was 1 million or greater. CONCLUSION: Using multiple logistic regression analysis, age of the women and serum oestradiol level had the maximum power to predict pregnancy following ovarian stimulation and intrauterine insemination. PMID- 14530529 TI - Clinical significance and management of cervical atypical glandular cells of undetermined significance. AB - OBJECTIVES: To assess the clinical significance of a cervical cytological diagnosis of atypical glandular cells of undetermined significance and to formulate the most appropriate management guidelines for patients with such a diagnosis. DESIGN: Retrospective study. SETTING: Regional hospital, Hong Kong. PATIENTS: Seventy-two patients with diagnoses of atypical glandular cells of undetermined significance who were managed in a colposcopy clinic between January 1998 and December 1999. MAIN OUTCOME MEASURES: Age, cytological diagnoses of atypical glandular cells of undetermined significance and its subtypes, method of evaluation, final diagnosis, and outcome after 2 years. RESULTS: Atypical glandular cells of undetermined significance were diagnosed in 83 (0.4%) of 21 854 cervical smear samples taken during the 2-year study period. Follow-up data were available from 72 patients, whose mean age was 43 years (range, 22-69 years). Forty-three percent of these patients had significant diseases of the genital tract. Patients with the subtype diagnosis of atypical glandular cells of undetermined significance-favour neoplasia had the worst outcome, with 90% of patients having significant disease, followed by patients with atypical glandular cells of undetermined significance "not otherwise specified" (43%), and atypical glandular cells of undetermined significance-favour reactive (8%). CONCLUSION: Patients with atypical glandular cells of undetermined significance should be investigated early and thoroughly, because many of them will have premalignant or malignant disease. PMID- 14530530 TI - Sleeping energy expenditure in paediatric patients with obstructive sleep apnoea syndrome. AB - OBJECTIVE: To investigate sleeping energy expenditure in paediatric patients with obstructive sleep apnoea syndrome. DESIGN: Retrospective study. Setting. University teaching hospital, Hong Kong. PATIENTS AND METHODS: A retrospective analysis comparing sleeping energy expenditure in patients with confirmed obstructive sleep apnoea syndrome and control subjects matched for age, sex, and ideal weight for body height. Subjects were recruited from the Paediatric Chest Clinic and all had undergone overnight polysomnography and sleeping energy expenditure measurement by open-circuit indirect calorimetry using a metabolic cart and canopy system. The measurements were taken during slow-wave sleep. RESULTS: Twenty-four cases with obstructive sleep apnoea syndrome (apnoea hypopnoea index >5) and 23 control subjects were studied. Mean age and ideal weight for body height were 9.4 (standard deviation, 3.9) years and 152.5% (27.2%), respectively. Mean sleeping energy expenditure corrected for body weight for the cases and the control group were 44.83 (standard deviation, 10.49) Kcal/day and 40.71 (10.60) Kcal/day, respectively. Sleeping energy expenditure was not found to be associated with the severity of obstructive sleep apnoea syndrome (r=0.34, P=0.27). CONCLUSION: A trend towards greater sleeping energy expenditure was found in patients with obstructive sleep apnoea syndrome. Further studies on the metabolic aspects of this condition are required. PMID- 14530531 TI - Sudden cardiac death: prevention and treatment. AB - Sudden cardiac death remains a major health issue in western countries as well as in Hong Kong. Despite increasing knowledge of the mechanisms and risk factors of sudden cardiac death, methods for identifying high-risk candidates and predicting the efficacy of measures to prevent sudden cardiac death are still inadequate. A significant proportion of patients have known heart disease but are generally considered to be at low risk for this event. More efforts are needed to improve the success rate of out-of-hospital resuscitation through better warning systems, the use of amiodarone for refractory arrhythmias, and the widespread availability of automated defibrillation devices to allow early defibrillation. It is likely that these measures could increase the number of survivors following cardiac arrest. In survivors of sudden cardiac death episodes, treatment of the underlying cardiac disease, especially early revascularisation for myocardial ischaemia, is required. In the majority of patients, implantation of an implantable cardioverter defibrillator, with or without the use of an anti arrhythmic drug such as amiodarone, would then be used to maintain survival. Furthermore, for individuals at significant risk of sudden cardiac death, primary prevention of sudden cardiac death through the placement of an implantable cardioverter defibrillator is increasingly being used. PMID- 14530532 TI - Thrombotic thrombocytopenic purpura as a rare complication in childhood systemic lupus erythematosus: case report and literature review. AB - Thrombotic thrombocytopenic purpura is a rare but serious condition in childhood. It can be idiopathic or a complication of other diseases or drug therapy. We report on a 12-year-old Chinese girl who presented with fulminant systemic lupus erythematosus with progressive renal failure, pancytopenia, and cerebral dysfunction due to thrombotic thrombocytopenic purpura. The patient also had Pneumocystis carinii pneumonia, Pseudomonas septicaemia, and Herpes zoster infections as a result of immunosuppressive treatment. She responded to combined therapy with pulse methylprednisolone, cyclophosphamide, plasmapheresis, and intensive care support, and completely recovered renal and neurological function. A review of the English-language medical literature since 1968 identified 20 other paediatric cases of systemic lupus erythematosus and thrombotic thrombocytopenic purpura. Clinical features, treatment, and outcome of these cases are presented and discussed. Early recognition is important, and although plasmapheresis is not of proven benefit in severe cases of systemic lupus erythematosus, it is life-saving in lupus-related thrombotic thrombocytopenic purpura and must be instituted early to avoid a poor outcome. PMID- 14530533 TI - Magnetic resonance imaging in ischaemic heart disease. AB - Magnetic resonance imaging has an increasing role in the assessment of ischaemic heart disease. Its superb spatial and temporal resolution currently allows accurate assessment of cardiac function, regional wall motion, and the extent of myocardial infarction. Regional myocardial perfusion can also be assessed, most commonly by a first-pass technique. Non-invasive imaging of the coronary arteries by various magnetic resonance imaging techniques represents a major advance in recent years. In the foreseeable future, magnetic resonance imaging may become a single, comprehensive examination for the assessment of ischaemic heart disease. PMID- 14530534 TI - Cholestatic jaundice caused by sequential carbimazole and propylthiouracil treatment for thyrotoxicosis. AB - A 36-year-old Chinese man presented to the Queen Mary Hospital in August 1999 with a 2-week history of jaundice due to propylthiouracil treatment for thyrotoxicosis. He had previously received carbimazole but had developed an urticarial skin rash after 2 weeks of treatment. The patient developed liver failure and fulminant pneumonitis shortly after hospital admission. Despite receiving treatment with broad-spectrum antibiotics and intravenous immunoglobulin, he died 11 days after the onset of the respiratory symptoms. Postmortem examination using electron microscopy showed typical glycogen bodies within the cytoplasm of the hepatocytes, which corresponded to eosinophilic cytoplasmic inclusion bodies visible under light microscopy. Immunohistochemical studies of the inclusion bodies were positive for carcinoembryonic antigen and albumin, and negative for fibrinogen, complement protein C3, immunoglobulins G, M, and A, alpha-fetoprotein, and alpha-1-antitrypsin. This is the first report of a patient who received two sequential antithyroid drugs and developed predominate cholestasis with unique histological features. Extreme caution should be taken when a patient develops allergy to one type of antithyroid drug, because cross reactivity may develop to the other type. PMID- 14530536 TI - Acute-on-chronic subdural haematoma: a rare complication after spinal anaesthesia. AB - An 88-year-old woman with an undiagnosed chronic subdural haematoma underwent emergency repair of a femoral hernia under spinal anaesthesia. The patient complained of headache postoperatively, and a subsequent computed tomography brain scan showed an acute-on-chronic subdural haematoma, with midline shift and impending coning. The patient recovered completely after surgical decompression. The difficulty in diagnosing chronic subdural haematoma in the elderly patient with no history of trauma is discussed, along with the differential diagnosis of headache following spinal anaesthetic in this age-group. PMID- 14530535 TI - A case of partial trisomy 13 presenting with hyperinsulinaemic hypoglycaemia. AB - We report on a newborn baby with partial trisomy 13 who presented with multiple dysmorphic features and hyperinsulinaemic hypoglycaemia. Cytogenetic study on peripheral blood lymphocytes showed 47,XY,+mar in all cells analysed; fluorescent in situ hybridisation showed that the marker was solely derived from chromosome 13. The final karyotype was 47,XY,+del(13)(q14q32). Milk formula through a nasogastric drip and intravenous glucose infusion were given to prevent further hypoglycaemia. However, the baby developed occasional episodes of hypoglycaemia during bolus feeding. Hence, diazoxide was given, at a dosage of 10 mg/kg per day from day 24. Thereafter, no hypoglycaemic episodes were detected. Subsequent follow-up revealed satisfactory growth, global developmental delay, and left divergent squint. PMID- 14530538 TI - The importance of communication. PMID- 14530537 TI - Cambodia: an extended sabbatical. PMID- 14530539 TI - Treating severe acute respiratory syndrome with hyperimmune globulins. PMID- 14530540 TI - Travel-acquired infections in general. PMID- 14530541 TI - Indoor air pollution and acute respiratory infections. PMID- 14530542 TI - Immune function in children with classical phenylketonuria and tetrahydrobiopterin deficiencies. AB - BACKGROUND: An increased susceptibility to infections has been observed in some patients with phenylketonuria (PKU), which is not well known whether it is due to alterations of plasma essential amino acid concentrations or to some other factors. OBJECTIVE: This study is designed to establish B cell and T cell functions in 44 children with classical PKU and tetrahydrobiopterin (BH4) deficiencies and the effects of too high plasma phenylalanine (PA) concentrations (16.53 to 30.54 mg/dL) on the same parameters. DESIGN: B and T cell functions of 33 children with classical PKU (divided into two groups based on fasting mean plasma PA concentrations: Group-I = 20.9 +/- 3.7 mg/dL, Group-II = 3.8 +/- 1.02 mg/dL), and 11 children with BH4 deficiencies (Group III) were studied. The results were compared between the groups and referenced with previously reported values from healthy controls. RESULT: Delayed type skin hypersensitivity responses to purified protein derivative (PPD) in Group I and phytohaemagglutinin (PHA) in Group I, III were lower than the other groups and healthy controls. Plasma IgG and IgM concentrations of Group I was lower than the reference values. Although mean serum zinc and iron levels of all patients were lower than published values of healthy children, zinc and iron deficiencies in Group I, III were much more prominent as compared to Group II. CONCLUSION: The somewhat low plasma IgG concentrations in Group I may be related to the very high plasma PA levels, however the role of zinc deficiency as a causal factor can not be ruled out. BH4 metabolism defects do not appear to affect the same parameters. Impaired delayed skin hypersensitivity responses in Group I and III can be explained by severe serum zinc deficiency. In the light of this study, we conclude that in order to establish a causal relationship between PKU and immune functions, further studies need to be conducted after the correction of micro-nutrient status of such children. PMID- 14530543 TI - Intramuscular injection: review and guidelines. PMID- 14530544 TI - IAP and industry relationship guidelines 2003. PMID- 14530545 TI - Environmental health for practicing pediatricians. PMID- 14530546 TI - Outbreak of killer brain disease in children: mystery or missed diagnosis? PMID- 14530547 TI - Unrecognised ventriculitis/meningitis presenting as hydrocephalus in infancy. AB - Infantile hydrocephalus due to unrecognized neonatal-onset meningitis/ventriculitis, was studied retrospectively using 1991-1998 chart review. Seventy two patients with hydrocephalus were reviewed. Thirteen infants had hydrocephalus associated with active meningitis/ventriculitis which had remained unrecognized. Active meningitis/ventriculitis was confirmed by the finding of an abnormal lumbar and ventricular CSF with or without positive culture. All had perinatal risk factors and 10/13 had been given antibiotics in the postnatal period. 6/13 infants appeared to be well. The most common presentation was increasing head size. All lumbar and ventricular CSFs were abnormal and 10/13 had positive cultures as well. Imaging revealed hydrocephalus in all. The infants were treated with antibiotics for a mean of 32.8 days before VP shunting. 7/11 were severely disabled. Unrecognized active meningitis/ventriculitis is an important cause of infantile hydrocephalus. PMID- 14530548 TI - Social and economic impact of childhood asthma. AB - In order to assess the social, educational and economic impact in children with asthma and their families, we studied 162 children with bronchial asthma. The patients and their parents were interviewed to assess the restriction on various activities of the child and family, the impact on schooling and expenditure on therapy. One hundred and forty one (87%) children had either mild or moderate persistent asthma. Nearly two thirds of children had some restriction placed on their play activities because of asthma. Restrictions on other physical activities and social activities were reported in half the children. Children had absented from school for a median of 4 days in preceding 6 months. All these restrictions were more common in children with more severe disease and/or poor control of symptoms. The median monthly expenditure on child's medication was Rs. 333, i.e., about one third of monthly per capita income. Childhood asthma has significant adverse impact on child's daily activities, schooling, and family life and finances. PMID- 14530549 TI - Role of C-reactive protein in deciding duration of antibiotic therapy in neonatal septicemia. AB - In this study serum C-reactive protein (CRP) levels were used to evaluate the duration of antibiotic therapy in 50 consecutive neonates with suspected septicemia. In 44 percent of cases therapy was stopped on 3rd day, as CRP was normal. In 8 percent antibiotics could be stopped within 5-7 days as CRP values returned to normal and in 48 percent therapy was extended beyond 7th day, as CRP values were high or rising persistently. Negative predictive value of serial CRP was 100 percent in deciding duration of antibiotic therapy in suspected neonatal septicemia up to 7 days. The correlation between positive CRP, raised micro ESR and positive blood culture was significant (p < 0.005). PMID- 14530550 TI - Normal urinary calcium/creatinine ratios in Turkish children. AB - A random urine calcium/creatinine ratio (UCa/Cr) is of practical use in screening for hypercalciuria. However, due to worldwide variations, reference values for the pediatric population are not yet well established. Furthermore, no study has been conducted to establish normal UCa/Cr values in Turkish children. The objectives of the present study were to set normal values of random UCa/Cr by age in the pediatric population of Istanbul City. A total of 324 healthy children of both genders were enrolled in the study. They were divided into four age groups as follows: (1) < 7 months, (2) 8-18 months, (3) 19 months to 6 years, and (4) 7 14 years. A non-fasting random urine specimen from each subject was analyzed for calcium and creatinine. The median UCa/Cr values were 0.19, 0.20, 0.14 and 0.10 respectively. The data showed a strong inverse relationship with age. The age dependent 95th percentiles of UCa/Cr values were 0.76, 0.60, 0.69 and 0.24 respectively. The child's age and geographic location should be taken into consideration when interpreting UCa/Cr ratio. PMID- 14530551 TI - Fraser-Cryptophthalmos syndrome with cardiovascular malformations: a rare association. AB - Fraser-Cryptophthalmos syndrome is a multiple malformation disorder associated variably with cryptophthalmos (hidden eye), anomalies of the head, nose and ears; syndactyly, renal and genital malformations. In this report, we describe a case of Fraser syndrome with cardiovascular malformations: coarctation of aorta, an association not previously described. PMID- 14530552 TI - Limb body wall complex. AB - A case of Limb body wall complex is reported in a preterm, Low birth weight and stillborn neonate. The proposita had abdominal wall defect with evisceration of the organs into an amnio-peritonial sac, a short body stalk, torsion of spine with scoliosis and limb defects. Unusual associations present in this case were absent thymus, interstitial calcification in kidney and a history of exposure to birth control pills during first two months post-conception. PMID- 14530553 TI - Oral bisphosphonates in polyostotic fibrous dysplasia. AB - The bisphosphonates inhibit osteoclastic bone resorption and have been used as an intravenous infusion in fibrous dysplasia of the bone. Oral bisphosphonates are cheaper and also easy to administer. We report a case of polyostotic fibrous dysplasia in a three-year-old child who showed significant improvement in bone mineral density after treatment with oral alendronate. PMID- 14530554 TI - Preoperative aspiration for anterior mediastinal cyst with respiratory distress. AB - We report a 5-month-old boy who presented with an anterior mediastinal cystic teratoma. Pre-operative symptoms of respiratory distress secondary to airway obstruction were markedly reduced by percutaneous aspiration. Aspiration was also beneficial in easily ventilating the baby during the surgical procedure. PMID- 14530555 TI - Lupus vulgaris. PMID- 14530556 TI - Candida parapsilosis on peripheral blood smear. PMID- 14530557 TI - Addison's disease. PMID- 14530558 TI - Antiphospholipid antibody syndrome as a cause of Budd-Chiari syndrome. PMID- 14530559 TI - Rickets in walled city of Delhi. PMID- 14530560 TI - Oral vitamin-K administration at birth. PMID- 14530561 TI - Hepatitis A: an unusual presentation. PMID- 14530562 TI - Massive CSF varicosity as a presenting feature of VP shunt disconnection. PMID- 14530563 TI - Inguinal hernias in babies. PMID- 14530564 TI - "Air entry"--an adventitious entry in respiratory system examination. PMID- 14530565 TI - Posterior urethral valves and anorectal malformation: a rare association. PMID- 14530566 TI - Comparison of HemoCue method with the cyanmethemoglobin method for estimation of hemoglobin. PMID- 14530567 TI - Combined Down and Klinefelter syndrome. PMID- 14530568 TI - Effect of vitamin A supplementation on childhood morbidity, mortality. PMID- 14530569 TI - Modified Ventilatory Index (MVI) in newborns with congenital diaphragmatic hernia (CDH). PMID- 14530570 TI - Brief profile of an epidemic of acute encephalopathy in Western Uttar Pradesh. PMID- 14530571 TI - Molecular markers of brain damage: current state and future perspectives. PMID- 14530572 TI - Proteins in cerebrospinal fluid and blood: barriers, CSF flow rate and source related dynamics. AB - Cerebrospinal fluid (CSF) routine analysis for diagnosis of neurological diseases is based on the concepts for discrimination of blood-derived and brain-derived immunoglobulin fractions in CSF. The actual molecular flux/CSF flow theory of the blood/CSF barrier function, which founded the hyperbolic discrimination lines in quotient diagrams, is derived from the laws of molecular diffusion combined with CSF flow rate. It emerged from this theory that the decrease of CSF flow rate is sufficient to explain quantitatively the increase of CSF protein concentrations as observed in many neurological diseases. With this concept of CSF flow rate as the modulator of the normal and pathological blood-CSF barrier function, we got for the first time a theoretical frame work to explain also quantitatively the dynamics of brain-derived proteins and their source related (neurons and glial cells or leptomeningal cells) differences. The review of the anatomical, physiological and biophysical knowledge points to the new interpretations: The changing albumin quotient is an indicator of changing CSF flow rate and not for a morphological "leakage" of the blood-brain barrier. As an application of these concepts the dynamics of brain-derived molecules in blood are discussed with two examples: beta trace protein, flowing with CSF into venous blood, and neuron specific enolase, passing from tissue into blood the opposite direction of serum proteins, again a gradient-dependent protein diffusion across the intact blood vessel wall. PMID- 14530573 TI - The Janus face of glial-derived S100B: beneficial and detrimental functions in the brain. AB - Glia play active and vital roles in the CNS, including regulation of brain development, maintenance of cellular homeostasis and reparative responses to acute and chronic neurologic insults. However, in pathological conditions, glial function may be dysregulated resulting in enhanced neuroinflammation and further neurologic injury. The fundamental concept that aberrant regulation of a normally beneficial process may instead result in pathology is exemplified by the properties of the glial-derived protein, S100B. In the developing brain and following acute glial activation in response to injury, S100B acts as a neurotrophic factor and neuronal survival protein. In contrast, overproduction of S100B by activated glia can lead to exacerbation of neuroinflammation and neuronal dysfunction. This duality of S100B actions, both reparative and degenerative, is consistent with the compelling clinical evidence of the association between increases in S100B levels and outcome following brain injury. S100B levels generally correlate with the severity of damage and may have predictive value for adverse neurological outcomes. The cumulative data support the potential of S100B as a biomarker for brain damage, implicate glial activation as a possible treatment target in acute and chronic CNS disorders, and highlight the dual role of glia in the reparative and pathologic responses to neurologic injury. PMID- 14530575 TI - Molecular markers of brain damage--clinical and ethical implications with particular focus on cardiac arrest. AB - Although 25-50% of patients suffering from cardiac arrest can be stabilised haemodynamically, the hospital discharge rate is only 2-14%. One of the major causes of this discrepancy is persistent brain damage. Studies to assess the prognostic value of early prediction of neurologic and overall outcome in patients with cardiac arrest have not yet produced precise and generally accepted diagnostic rules. As apparative diagnostic methods often fail to predict neurologic outcome, the role of molecular markers has come a focus of common interest for early outcome prediction. This systematic review article aims to give an overview on the most important molecular markers for neurologic and overall outcome prediction and outline the advantages, clinical implications and ethical issues in patients undergoing cardiopulmonary resuscitation after cardiac arrest. For this purpose, the traditional marker for brain damage, the neuron specific enolase, a gamma gamma isomer of enolase and cytoplasmatic enzyme of glycolysis, and the astroglial protein S100, a calcium-binding protein regulating neuronal differentiation, outgrowth, and apoptosis, are analysed and their role discussed as a marker for brain damage in general and recovery after cardiopulmonary resuscitation following cardiac arrest. Neuron-specific enolase has been investigated as a neuro-marker after brain damage and for outcome prediction in unconscious patients. Whereas the protein S100 has proven to be a good marker for neuronal damage after isolated brain injury, its role in cardiac surgery is not as clear: at least, in the early postoperative phase S100 is not a sole marker for neurologic damage, as release of S100 from cardiac tissue and other sources has also been demonstrated. However, the persistent elevation of S100 after cardiac surgery is specific for neurologic impairment. Most interestingly, after cardiac arrest the protein S100 has shown to be a good survival marker for overall outcome prediction. Although it cannot be absolutely determined whether cerebral or cardiac release of S100 is predominant in this clinical setting, recent studies have revealed that S100 serum levels are a useful diagnostic tool for outcome prediction. In contrast, after cardiac arrest serum levels of protein S100 did not reach a 100% specificity and sensitivity in clinical studies, and, therefore, elevated S100 in these patients has to be interpreted with caution. Nonetheless, low S100 serum levels have been correlated with good outcome and, therefore, even if all other diagnostic tests indicate poor outcome, all therapeutic efforts must be undertaken, as no single study has shown that normal S100 serum levels were associated with poor prognosis. PMID- 14530574 TI - Peripheral markers of brain damage and blood-brain barrier dysfunction. AB - PURPOSE: Occurrence of brain damage is frequently associated with abnormal blood brain barrier (BBB) function. Two brain-specific proteins, S100beta and neuron specific enolase (NSE) are released systemically in a variety of neurological diseases, but S100beta levels sometimes rise in the absence of neuronal damage, suggesting that S100beta is a marker of BBB rather than neuronal damage. METHODS: We measured both proteins in the serum of patients undergoing iatrogenic BBB disruption with intrarterial mannitol, followed by chemotherapy. RESULTS: Serum S100beta increased significantly after mannitol infusion (p<0.05) while NSE did not. Furthermore, in a model of intracerebral hemorrhage, S100beta increases in CSF did not lead to serum changes at a time when the BBB was intact. Modeling of S100beta release from the CNS suggested that low (<0.34 ng/ml) serum levels of S100beta are consistent with BBB opening without CNS damage, while larger increases imply synthesis and release from presumable damaged glia. CONCLUSIONS: Thus, S100beta in serum is an early marker of BBB openings that may precede neuronal damage and may influence therapeutic strategies. Secondary, massive elevations in S100beta are indicators of prior brain damage and bear clinical significance as predictors of poor outcome or diagnostic means to differentiate extensive damage from minor, transient impairment. PMID- 14530576 TI - Release patterns of astrocytic and neuronal biochemical markers in serum during and after experimental settings of cardiac surgery. AB - OBJECTIVE: Brain injury and altered psychomotor development in infants, children and adults after cardiac surgery using cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest (DHCA) is still a matter of concern. Early diagnosis and identification of brain injury that has occurred or is ongoing by measurement of biochemical markers in serum may have diagnostic and prognostic value. The aim of the experimental studies in an animal model was therefore to investigate the release patterns of astroglial and neuronal markers in serum and to determine the morphological and immunohistochemical changes in the brain of animals undergoing similar perfusion conditions of CPB and a period of DHCA. METHODS: Fourteen New Zealand rabbits, (weight, 3.1 +/- 0.25 kg) were anesthetized, intubated and mechanically ventilated. Four animals were sham operated and served as controls. After median sternotomy the animals were connected to CPB by cannulation of the aorta and right atrium. Full flow CPB (200 250 ml/kg/min) was initiated to achieve homogeneous systemic cooling. Circulatory arrest of 60 minutes was induced when rectal and nasopharyngeal temperature of 14 degrees C was achieved. After rewarmed reperfusion and establishment of stable cardiac ejection the animals were weaned from CPB and monitored for 6 hours. Then the animals were killed, the brain was immediately removed and cut in standardized sections. These were fixated, embedded in paraffin and stained for further quantitative histological studies. In the brain astrocyte reactivity for S-100B was assessed immunocytochemically (DPC Immustain Los Angeles, USA). Monoclonal mouse anti-human neurospecific enolase (NSE) antibody was used for the localization of NSE in the fixed and paraffin embedded brain (NSE-DAKO, H14). The concentrations of S-100B protein and neurospecific enolase (NSE) in the serum were analyzed using a commercially available immunoluminometric assay (LIA-mat, Sangtec 100, Byk-Sangtec). Immunospecific monoclonal anti-parvalbumin antibody was used for the detection of parvalbumin in the brain. Serum concentrations of parvalbumin were analyzed using a newly developed ELISA method. RESULTS: In all experimental animals a significant increase of the serum concentration of the astroglial protein S-100B was found immediately after reperfusion and the termination of CPB. In contrast the serum levels of the neuronal proteins parvalbumin and NSE were not increased, but rather decreased. Light microscopy and electron microscopy revealed perivascular astrocytic swelling and minor neuronal cell injury. In comparison to the sham operated animals, increased immunohistochemical staining of S-100B was found. This increased reactivity of S100B antibody was found in the astrocytic processes with immediate connection to the perivascular space and around the perivascular oedema. The immunocytochemical stainings for NSE and parvalbumin in the neuronal cells was not different from that of sham-operated animals and indicated well preserved neurons. PMID- 14530577 TI - S100B and cardiac surgery: possibilities and limitations. AB - Serum determinations of the glial protein S100B has been found to correlate with brain damage after cardiac surgery. Forty-eight hours and later after surgery, increased S100B levels correlates with the presence of brain infarction, and the extent of infracted brain tissue. S100B at this time-point has been shown to predict long-term outcome, higher S100B levels correlated with decreased survival. Early levels (2-8 hours after surgery) of S100B have shown disparate results when trying to correlate it with postoperative cognitive decline. One reason for the lack of strong correlation could be the contamination of S100B from shed blood the first hours after surgery. PMID- 14530578 TI - Serum S-100B protein as a molecular marker in severe traumatic brain injury. AB - PURPOSE: There is growing evidence that S-100B protein may be used as a novel biochemical marker of brain cell damage, measured by a simple blood test. In this paper, we summarize the current knowledge about S-100B serum measurements in severe head injury and address actual controversies. METHODS: The material of the present analysis consists of a MEDLINE literature search for S-100 and severe head injury from 1966 to 2003. Studies of S-100B in severe head injury were reviewed for their information about the potential clinical value of this marker. RESULTS: A total of 18 clinical studies were identified and reviewed. Peak values of serum S-100B were demonstrated to have the highest correlation to any endpoint parameter. Admission values also have a significant correlation to injury severity and outcome but show a highly time dependent temporal course. There is extracranial release of S-100B in multitrauma patients in the first 48 hours, but the impact is limited as many studies have found a clear relationship between S 100B and injury severity, imaging findings and outcome. CONCLUSION: S-100B belongs to a new generation of molecular serum markers of brain damage. These markers will have potential as a surrogate outcome marker or monitoring parameters for both clinical and experimental settings. PMID- 14530579 TI - Biochemical serum markers for brain damage: a short review with emphasis on clinical utility in mild head injury. AB - PURPOSE: To provide an overview of clinical research on the use of biochemical serum markers for traumatic brain injury (TBI) in the evaluation of patients with mild head injuries (MHI). METHODS: The MEDLINE database was searched for publications on biochemical serum markers of TBI until August 2002. Clinical studies addressing their use in MHI were reviewed. RESULTS: Desirable characteristics for biochemical serum markers of TBI were identified. Creatine kinase isoenzyme BB (CK-BB), neuron specific enolase (NSE) and S-100B protein have been most extensively studied. The sensitivity and specificity of CK-BB is inadequate for use as an indicator of traumatic brain injury. Serum levels of NSE do not correspond to the amount of TBI, probably because of its long (20 h) half life. S-100B serum levels are correlated to both clinical measures of injury severity, neuroradiological findings and outcomes in several studies from different authors. CONCLUSION: Currently, S-100B protein is the most promising marker for evaluation of TBI in patients with MHI. PMID- 14530580 TI - Brain derived proteins as markers of acute stroke: their relation to pathophysiology, outcome prediction and neuroprotective drug monitoring. AB - PURPOSE: Ischemic stroke is associated with a variety pathophysiological changes affecting both glial and neuronal brain tissue. These changes are mirrored in the release of specific proteins into peripheral blood. Neurone-specific enolase (NSE), protein S100B and glial fibrillary acidic protein (GFAP) are those proteins investigated most often as peripheral surrogate markers of brain damage after stroke in humans. METHODS AND RESULTS: In the present article we present data which show that the release patterns of neuronal and glial tissue derived proteins after acute stroke are associated with the neuroradiological and neurobehavioral consequences of ischemic brain lesions and, additionally, may be useful in short-term outcome prediction. Kinetics of protein serum concentrations, however, are highly dependent on subtype of stroke lesions and do not merely reflect the degree of brain damage. They rather express complex neuronal-glial interactions as a (patho-)physiological consequence of ischemic brain lesions. We further demonstrate that S100B release patterns do reflect successful neuroprotective drug treatment with recombinant human erythropoietin (EPO) in acute stroke patients. CONCLUSIONS: The analysis of poststroke serum concentrations of glial tissue derived proteins might be a promising strategy to monitor and evaluate neuroprotective approaches in stroke treatment. PMID- 14530581 TI - Differential diagnosis of neurodegenerative diseases with special emphasis on Creutzfeldt-Jakob disease. AB - In principle, two research approaches can be considered for the laboratory diagnosis of transmissible spongiform encephalopathies (TSE): (i) the direct detection of PrPSc and (ii) the detection of surrogate markers in biological materials that show an altered pattern of expression in early stages of the disease or are used in the differential diagnosis of other dementias and thus enable an exclusion diagnosis. This review concentrates on the second approach. It was shown that a single determination of just a few markers (tau-protein, S 100B, 14-3-3-protein) was already sufficient to achieve a high degree of diagnostic certainty in the diagnosis of CJD. On the basis of the available data, it is to be expected that a combination of these markers will bring improved diagnostic strength with regard to the differential diagnosis of dementias as a whole. This especially applies to some of the subtypes of CJD and Alzheimer's dementia (AD). PMID- 14530583 TI - Gender differences in a sample of vocational rehabilitation clients with TBI. AB - Because traumatic brain injury affects between 1.5 and 2 million individuals per year and results in long term vocational and financial difficulties, there is growing interest in determining those factors that predict successful outcomes for specific groups of individuals with TBI. An NIH consensus panel on TBI has suggested that women are one group that needs more attention, particularly given the studies indicating that men and women experience different cognitive [14], emotional [19], and vocational outcomes following TBI [5]. The current study evaluated differences in injury severity, demographics, neuropsychological abilities, and vocational and financial outcomes for 78 persons with TBI (55 male, 23 female) who received services from a state Vocational Rehabilitation Division (DVR). Despite similar injury severity, neuropsychological and demographic characteristics, more men (43.6%) received Maintenance services from MO-DVR than women (21.7%). Of note, only 4.4% of the women were successfully employed through DVR, compared to 23.6% of the men. In addition, 73.9% of the women had services terminated after being accepted by DVR but before services were initiated, compared to 56.4% of the men. The significance of these results is discussed, as are the limitations of the current project. PMID- 14530584 TI - Rural/urban differences in vocational outcomes for state vocational rehabilitation clients with TBI. AB - OBJECTIVE: To evaluate differences in demographics, injury severity, and vocational outcomes for persons with TBI based on rural vs. urban residency. PARTICIPANTS: 78 individuals with TBI (28 from rural counties, 50 from urban counties) who requested services from the Missouri Division of Vocational Rehabilitation (VR) over a two year period. MEASURES: Demographics (i.e., age, race, education), injury severity (i.e., loss of consciousness, post traumatic amnesia, length of hospitalization, neuropsychological test scores), VR services provided (e.g., transportation, maintenance, on-the-job training, etc.), and VR outcomes (successfully vs. unsuccessfully employed; cost per case). PROCEDURE: All participants completed a standard neuropsychological evaluation and completed VR services (i.e., were followed from enrollment to case closure). Rural and urban residency was determined using U.S Office of Management and Budget definitions of metropolitan and non-metropolitan areas. ANALYSIS: Chi-squares, Fisher's Exact tests, Wilcoxon Rank Sums test, and MANOVAs. RESULTS: Few if any differences were found between the groups in demographics (i.e., more African Americans in urban areas), injury severity (i.e., more rural residents with multiple TBIs), or neuropsychological test scores. However, individuals from urban areas received significantly more maintenance funds (46% vs. 21%), transportation services (36% vs. 11%), and on-the-job training (28% vs. 7%), and had more spent on them ($1,816 vs $1,242). Although statistically non-significant (p < 0.15), 24% of individuals from urban areas were successfully employed at VR case closure, compared to only 7% of individuals from rural areas. CONCLUSIONS: Individuals with TBI from rural and urban settings have generally similar demographic, injury severity, and neuropsychological abilities, although they appear to differ in terms of vocational outcomes and number of VR services received, possibly related to limited availability of resources in rural areas. PMID- 14530585 TI - Loxapine versus olanzapine in the treatment of delirium following traumatic brain injury. AB - The use of typical neuroleptics has always been the mainstay of treatment for delirium following traumatic brain injury (TBI). Given the recent application of atypical neuroleptics to various psychiatric conditions formerly treated with typical neuroleptics, one questions whether this new class of drugs is superior to its predecessor in treating delirium post-TBI. We present a case of one patient with TBI-induced delirium where in fact the use of the typical mid potency neuroleptic, loxapine, appeared to have a better clinical effect over the atypical neuroleptic, olanzapine. PMID- 14530586 TI - Impairments in activities of daily living in Parkinson's disease: implications for management. AB - Parkinson's disease (PD) is a neurodegenerative disorder with progressive impairment in motor and cognitive functioning. A review of past research shows that individuals experience not only physical discomfort brought on by the illness, but also considerable psychological distress. Increased dependency on a caregiver due to impairments in activities of daily living (e.g., walking, eating, dressing) may bring on feelings of being a burden and strain relationships. However, since PD is a chronic illness, it necessitates family caregiving throughout the life-span. This has implications for the family members who need to redefine their roles within the family to manage the circumstances brought on by the illness. Such strains may include finances, threats to employment, social isolation or health strain. Thus, it becomes imperative to provide unrelenting support to the affected family and individual, to enable them to refocus and learn to manage new illness-related, task-oriented, and affective roles. This enhances the integrity and self-esteem of the patient and helps the family's adjustment to illness of a member. This paper outlines, the role of mental health professionals in providing counseling and psychoeducational approaches to helping patients and their families. PMID- 14530587 TI - A home-based, self-administered stimulation program to improve selected hand functions of chronic stroke. AB - OBJECTIVES: To test if a combined stimulation-training program can improve selected hand functions and impairments of chronic stroke survivors. DESIGN: Pretest-Posttest, multi-site 5-week training program. Stroke survivors (N = 77) with chronic upper limb paresis completed a home-based stimulation program combining activation of the wrist-fingers flexors and extensors with functional grasp, hold and release training. Subjects trained 2-3 times each day, 7 days a week. Outcome measures included: the Jebsen-Taylor simulated feeding (S-feed); light object lift (J-T light); heavy object lift (J-T heavy); Box and Blocks test (B+B); Nine-Hole Peg (9-HP); Ashworth scale (Spasticity); Visual analog scale-VAS (Pain). Paired t-tests (alpha < 0.01) were performed on each study outcome. RESULTS: Simulated feeding time decreased from 39.1 +/- 30.9 sec to 25.5 +/- 23.3 sec (34.8% improvement). The task time of the J-T light decreased by 13.3 sec and the J-T heavy by 11.5 sec (44.9% and 40.9% improvement respectively). The number of blocks moved increased from 19.4 +/- 11.6 to 24.5 +/- 12.5 (26.3% improvement) and the time to complete the 9-HP decreased from 178.8 +/- 170.8 to 105.0 +/- 117.1 sec (58.7% improvement). Mean reduction of spasticity was 0.87 and 0.78 points at the elbow and wrist respectively. Patients with persistent pain (N = 33) reported mean reduction from 3.5 +/- 2.5 to 1.9 +/- 1.8. CONCLUSIONS: Five weeks of daily home training with a task-specific stimulation program improved selected hand functions and upper limb impairments associated with chronic post stroke paresis. PMID- 14530588 TI - Repetitive visual stimulation: a neuropsychological approach to the treatment of cortical blindness. AB - Cortical visual disturbances can occur after traumatic or ischemic brain lesion. Patients mostly suffer from hemianopia. Different treatment approaches in patients with hemianopia are being debated with respect to their effectiveness. For more severely disabled patients with cortical blindness or residual rudimentary vision (RRV) no systematic therapeutic approaches have been reported. In a case study the positive effects of a recently developed repetitive photic stimulation therapy in a patient with RRV after a bioccipital ischemic infarction are presented. The application of this new therapy over several months, supported by treatment with amphetamines led to a statistically significant improvement of different visual functions and a reoccurrence of visual abilities important in daily life. The pathophysiological basis and possible neurorehabilitative consequences that arise from these results are discussed with respect to similar findings in animal experiments. PMID- 14530589 TI - The ICF: Applications of the WHO model of functioning, disability and health to brain injury rehabilitation. AB - The traditional disease and diagnostic medical model is not always useful to brain injury professionals who need to describe, measure, and compare deficits associated with neurologic insult. Professionals in neurorehabilitation are in need of new systems that will assist them in identifying impairments and areas of intervention. The aim of this article is to present the International Classification of Functioning, Disability and Health (ICF), and its applications to brain injury rehabilitation. This taxonomy, developed by the WHO, allows the classification and assessment of functioning and disability in everyday activities and social involvement for individuals with medical conditions. Multi disciplinary teams from 65 countries have collaborated in the development of the ICF to develop a tool that serves different purposes and disciplines with high trans-cultural validity. It can be of great value for professionals working in the field of brain injury who need to describe and quantify in detail neurocognitive, emotional, and sensory-motor functions as well as their impact on activities and participation in life situations. Its applications also extend to the domains of epidemiology, public health and public policy. PMID- 14530590 TI - BalanceReTrainer: a new standing-balance training apparatus and methods applied to a chronic hemiparetic subject with a neglect syndrome. AB - In this paper we present a mechanical apparatus and methods named BalanceReTrainer for standing-balance training in neurologically impaired individuals. BalanceReTrainer provides an impaired individual with a fall-safe balancing environment, where the balancing efforts of a standing individual are augmented by stabilizing forces acting at the level of pelvis in the sagittal and frontal planes of motion, assisting the balancing activity ankle and hip muscles and at the level of shanks, assisting the knee extensor muscles. A range of different levels of supporting forces is generated by passive, compliant means. Additionally, movement in the sagittal and frontal planes, acquired by transducers is fed to an electronic interface which transforms the current inclinations into a computer mouse signals, which are interfaced to a personal computer (PC) where balance training and evaluation program is running. The level of stiffness support and level of difficulty of computer task can be selected according to current balancing abilities of the impaired individual. We further present results of a case study where an ambulatory chronic hemiparetic subject with neglect syndrome received ten days of balance training on BalanceReTrainer. Biomechanical evaluation of weight-shifting activity before and after treatment shows a substantial functional improvement. PMID- 14530591 TI - Life expectancy for children with cerebral palsy and mental retardation: implications for life care planning. AB - OBJECTIVES: Physicians may be asked by attorneys or other patient advocates to help plan for the long-term needs of children with cerebral palsy (CP) and developmental disability (DD). The first step in such planning is to thoroughly examine the literature dealing with life expectancy in these populations. This review paper comprehensively reviews the literature relating to survival in children with cerebral palsy and developmental disability. STUDY SELECTION: A Medline data search was completed using the terms cerebral palsy, life expectancy, survival, as well as other pertinent terms. Further articles were gleaned from bibliographies of pertinent literature. DATA SYNTHESIS: Certain key disabilities can be used to accurately predict life expectancy in children with cerebral palsy and mental retardation. These include: (1) presence and severity of mental retardation, (2) inability to speak intelligible words, (3) inability to recognize voices, (4) inability to interact with peers, (4) severity of physical disability, (5) use of tube feeding, (6) incontinence, and (7) presence and severity of seizures. CONCLUSIONS: Literature review definitively shows that children with CP and DD have a diminished life expectancy, which can be assessed based on simple clinical examination findings. PMID- 14530592 TI - Treatment benefit correlates with increase of daily drug costs in Parkinson's disease clinics. AB - BACKGROUND: There is a move towards treatment of Parkinson's disease (PD) patients in specialized units, however, data on the outcome and on daily antiparkinsonian drug costs are rare. OBJECTIVE: The objective of this study was to elucidate relationships between costs of drug treatment and efficacy of drug titration in a PD clinic. SUBJECTS AND METHODS: We calculated costs of drug therapy and scored severity of PD of 63 consecutively referred in-patients initially and at the end of their hospital stay under standardized conditions. RESULTS: Titration of antiparkinsonian drugs significantly induced a decrease of PD symptoms and an increase of daily drug costs. There were significant correlations between the degree of (i) improvement of the UPDRS score, (ii) increase of dopaminergic agents and (iii) change of corresponding daily antiparkinsonian drug costs. CONCLUSION: Our results demonstrate the effectiveness of treatment in PD clinics, which results in increased daily antiparkinsonian drug costs due to elevated dopaminergic substitution. PMID- 14530594 TI - Antioxidant effects of tetrahydro-beta-carboline derivatives identified in aged garlic extract. AB - 1,2,3,4-Tetrahydro-beta-carboline derivatives (THbetaCs) are formed through Pictet-Spengler chemical condensation between tryptophan and aldehydes during food production, storage and processing. In the present study, in order to identify the antioxidants in aged garlic extract (AGE), we fractionated it and identified four THbetaCs; 1-methyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acids (MTCC) and 1-methyl-1,2,3,4-tetrahydro-beta-carboline-1,3-dicarboxylic acid (MTCdiC) in both diastereoisomers using liquid chromatography mass spectrometry (LC-MS). Interestingly, these compounds were not detected in raw garlic, but the contents increased during the natural aging process of garlic. In in vitro assay systems, all of these compounds have shown strong hydrogen peroxide scavenging activities. (1S, 3S)-MTCdiC was found to be stronger than the common antioxidant, ascorbic acid. MTCC and MTCdiC inhibited AAPH-induced lipid peroxidation. Both MTCdiCs also inhibited LPS-induced nitrite production from murine macrophages at 10-100 microM. Our data suggest that these compounds are potent antioxidants in AGE, and thus may be useful for prevention of disorders associated with oxidative stress. PMID- 14530593 TI - Membrane-rigidifying effects of anti-cancer dietary factors. AB - Since several anti-cancer drugs interact with cell membrane lipids, the effects of anti-cancer dietary factors on liposomal membranes with different lipid composition were comparatively studied by measuring fluorescence polarization. Fluidity was imparted on both hydrophobic and hydrophilic regions of lipid bilayers by decreasing cholesterol and increasing unsaturated phosphatidylcholine in membranes. At 0.625-10 microM, (-)-epigallocatechin gallate, genistein, apigenin, resveratrol and a reference anti-cancer drug, doxorubicin, rigidified the tumor cell model membranes consisting of 20 mol% cholesterol and 80 mol% phosphatidylcholine with the acyl chain 18:1/16:0 ratio of 1.0, but not daidzein. They were more effective on the membrane core than the membrane surface. Quercetin showed a biphasic effect on the hydrophobic regions of membrane lipid bilayers to rigidify above 5 microM and fluidize below 2.5 microM. In contrast, anti-cancer dietary factors and doxorubicin were not or much less effective in rigidifying the normal cell model membranes consisting of 40 mol% cholesterol and 60 mol% phosphatidylcholine with the acyl chain 18:1/16:0 ratio of 0.5. The membrane-rigidifying effects were greater depending on a decrease of the cholesterol/phosphatidylcholine ratio and an increase of the phosphatidylcholine unsaturation degree. Membrane-active dietary factors and doxorubicin inhibited the growth of mouse myeloma cells at 10-100 microM, while the growth inhibition by membrane-inactive daidzein was relatively weak. Anti-cancer dietary factors appear to act on more fluid membranes like tumor cells as well as doxorubicin to induce rigidification, especially in the hydrocarbon core of membrane lipids, which is determined by the composition of cholesterol and unsaturated phospholipids. PMID- 14530595 TI - Characteristic rat tissue accumulation of nobiletin, a chemopreventive polymethoxyflavonoid, in comparison with luteolin. AB - Nobiletin (NOB), a polymethoxyflavonoid, is an effective anti-inflammatory and chemopreventive phytochemical found in citrus fruits. We compared the absorption and metabolism characteristics of NOB with those of luteolin (LT) in male SD rats. Each flavonoid (67.1 micromol/kg of body weight) was given separately by gastric intubation, and then concentrations were measured at 1, 4, and 24 hours after administration. In the digestive organs, NOB showed a notable tendency for localizing into the mucous membrane and muscularis from 1 to 4 hours, in contrast to LT, though both NOB and LT were completely excreted within 24 hours. Further, significant amounts of NOB were detected in the whole liver and kidney specimens, whereas LT accumulation was slight. Although serum concentrations of NOB from 1 to 4 hours were comparable to those of LT, urinary concentrations of LT were significantly higher from 4 to 24 hours. Following glucuronidase/sulfatase treatments of urinary materials, we detected 3 types of mono-demethylated NOB, including 3'-demethyl-NOB, and two di-demethylated types, as well as 3'-demethyl NOB alone in serum samples using liquid chromatography-mass spectral analysis. Our results suggest that the metabolic properties of polymethoxyflavonoids are distinct from those of other general flavonoids, because of their wide distribution and accumulation in tissue. PMID- 14530596 TI - Impact of water-dispersible beadlets as a vehicle for the delivery of carotenoids to cultured cells. AB - Water-dispersible beadlets of carotenoids were used as supplements for human umbilical vein endothelial cells (HUVECs), human peripheral blood mononuclear cells (PBMCs) and human monocytes. Stability, cellular association and cytotoxicity of the carotenoid beadlets were compared with carotenoids delivered with tetrahydrofuran (THF). Incubations with lycopene, beta-carotene, lutein and astaxanthin dissolved in THF resulted in a lower stability in the medium, lower cellular association, and a higher standard deviation. Beadlets provided 60, 4, 6, and 2 times greater accumulation of lycopene, beta-carotene, lutein and astaxanthin, respectively, by PBMCs than THF. The cellular association of carotenoids delivered by THF seems to be more carotenoid-specific than when carotenoids are delivered by beadlets. After 48 h of incubation under cell culture conditions all of the four carotenoids (1 microM) delivered by beadlets to the medium showed a reduction less than 30%. In addition, no cytotoxic effect of the carotenoid beadlets or the vehicle alone was detected in a concentration range of 0.5-5 microM. The results show that beadlets are a non-toxic vehicle for supplementing and stabilizing carotenoids in culture media offering a reasonable compromise in term of cell accumulation efficiency. PMID- 14530597 TI - Antioxidant effects of alpha- and gamma-carboxyethyl-6-hydroxychromans. AB - Carboxyethyl-6-hydroxychromans (CEHC), the major metabolites of both tocopherols (Toc) and tocotrienols (Toc-3), have been found in human plasma. In the present study, the antioxidant properties of alpha- and gamma-CEHC were measured and compared with alpha- and gamma- tocopherols. Following results were obtained: (1)alpha- and gamma-CEHC have the same reactivities toward radicals and exert the same antioxidant activities against lipid peroxidation in organic solution as the corresponding parent tocopherols respectively; (2) the partition coefficient decreased in the order alpha-Toc (3.36) > gamma-Toc (3.14) > alpha-CEHC (2.26) > pentamethyl-6-chromanol (1.92) > gamma-CEHC (1.83) > 0 > Trolox (-0.97); (3) alpha- and gamma-CEHC scavenge aqueous radicals more efficiently but they inhibit the lipid peroxidation within the membranes less efficiently than the corresponding alpha- and gamma-Toc, respectively; (4) alpha-CEHC inhibits the oxidation synergistically with ascorbate; and (5) alpha- and gamma-CEHC reduce Cu(II) to give Cu(I) and corresponding quinones as major product, but the prooxidant effect of CEHC in the presence of cupric ion was small. These results imply that CEHC may act as an antioxidant in vivo especially for those who take tocopherol supplement. PMID- 14530598 TI - Amelioration of cisplatin toxicity by a fermented grain food product. AB - The most noticeable hypothesis regarding the pathogenesis of cisplatin toxicity, seen mainly in kidney and intestine, is oxidative stress, an imbalance between free-radical generating cisplatin and radical scavenging systems. This paper describes the role of the antioxidant system in cisplatin-induced toxicity and the protective effect by a processed grain food (Antioxidant Biofactor: AOB), which has been shown to exhibit strong antioxidant activity. Male Fischer 344 rats were used. They were pre-fed either a basal diet (control, 15 g/day) or the diet supplemented with AOB to provide 6.5% or 20% of total diet throughout the experiment. Cisplatin (5 mg/kg, i.v.) was administered at the start of the experiment, and the animals were sacrificed 5 days later. Blood urea nitrogen (BUN) and plasma creatinine, NO2(-) and NO3(-) (NOx) were determined from the plasma. The levels of 4-hydroxy-2-nonenal (a lipid peroxidation product), 8 hydroxy-deoxyguanosine (8-OHdG, an oxidatively modified DNA adduct) and nitrotyrosine were histologically analyzed. The cisplatin administration resulted in a loss of body weight and elevations of BUN, serum creatinine and NOx levels, whereas AOB supplement reversed these effects. The severe morphological damages induced in the kidney and intestine by the cisplatin administration were markedly improved in the AOB group. The levels of lipid peroxidation, 8-OHdG, and nitrotyrosine all paralleled the morphological damage. The AOB effect was dose dependent. In conclusion, the present study suggests that certain food additives like AOB may be of benefit against the side effects of cisplatin. PMID- 14530599 TI - Magnetofection potentiates gene delivery to cultured endothelial cells. AB - Modification of cellular functions by overexpression of genes is increasingly practised for research of signalling pathways, but restricted by limitations of low efficiency. We investigated whether the novel technique of magnetofection (MF) could enhance gene transfer to cultured primary endothelial cells. MF of human umbilical vein endothelial cells (HUVEC) increased transfection efficiency of a luciferase reporter gene up to 360-fold compared to various conventional transfection systems. In contrast, there was only an up to 1.6-fold increase in toxicity caused by MF suggesting that the advantages of MF outbalanced the increase in toxicity. MF efficiently increased transfection efficiency using several commercially available cationic lipid transfection reagents and polyethyleneimine (PEI). Using PEI, even confluent HUVEC could be efficiently transfected to express luciferase activity. Using a green fluorescent protein vector maximum percentages of transfected cells amounted up to 38.7% while PEI without MF resulted in only 1.3% transfected cells. Likewise, in porcine aortic endothelial cells MF increased expression of a luciferase or a beta-galactosidase reporter, reaching an efficiency of 37.5% of cells. MF is an effective tool for pDNA transfection of endothelial cells allowing high efficiencies. It may be of great use for investigating protein function in cell culture experiments. PMID- 14530600 TI - Visualization of leukocyte transendothelial and interstitial migration using reflected light oblique transillumination in intravital video microscopy. AB - Dynamic visualization of the intravascular events leading to the extravasation of leukocytes into tissues by intravital microscopy has significantly expanded our understanding of the underlying molecular processes. In contrast, the detailed observation of leukocyte transendothelial and interstitial migration in vivo has been hampered by the poor image contrast of cells within turbid media that is obtainable by conventional brightfield microscopy. Here we present a microscopic method, termed reflected light oblique transillumination microscopy, that makes use of the optical interference phenomena generated by oblique transillumination to visualize subtle gradients of refractive indices within tissues for enhanced image contrast. Using the mouse cremaster muscle, we demonstrate that this technique makes possible the reliable quantification of extravasated leukocytes as well as the characterization of morphological phenomena of leukocyte transendothelial and interstitial migration. PMID- 14530601 TI - In situ analysis of coronary terminal arteriole diameter responses: technical report of a new experimental model. AB - INTRODUCTION: To date, investigation of coronary arteriole vasomotor activity has been limited to arterioles >30- 40 microm. Here, we introduce a new experimental model to allow for in situ microscopy of terminal coronary arterioles. METHODS: Rat hearts were perfused in a closed loop system (priming volume 20 ml) which was placed on a computer-controlled microscope stage. FITC-dextran and tetrodotoxin (TTX, 50 microM) were added. Tilting of the microscope by 90 degrees allowed for visual access to the ventricular surface. Arterioles were identified by the flow direction of fluorescent beads (1 microm). Images were recorded on video tape, and arteriole diameters were measured offline. Stability of the preparation and maintenance of coronary flow reserve were analyzed. Responses of coronary flow and arteriole diameters to the vasodilators papaverine and Na-nitroprusside were recorded. RESULTS: In TTX-arrested control hearts coronary flow and terminal arteriole diameters were stable for 2 h. Administration of papaverine and Na nitroprusside increased coronary flow from 6.4 +/- 0.7 to 13.3 +/- 1.3 ml/min, decreasing coronary resistance by 52 +/- 3%. Terminal coronary arteriole diameters increased from 12.0 +/- 0.9 to 13.6 +/- 1.0 microm, decreasing hindrance of this vessel segment by 45 +/- 11%. CONCLUSION: Preservation of coronary terminal arteriolar tone and adequate responsiveness to vasodilators in the TTX-arrested isolated heart were demonstrated. Thus, this model may serve to complement our understanding of coronary microvascular control mechanisms by extending observations to the terminal arteriolar bed. PMID- 14530602 TI - Genetic analysis of short stature. AB - Short stature is a major concern for patients and their parents, and represents a diagnostic challenge to the clinician. A correct diagnosis is of particular importance in view of the availability of effective, but costly, therapy in a small subset of cases. Many different genetic etiologies of short stature are known. Therefore, chromosome as well as molecular analysis are requisite diagnostic investigations in children with short stature. Particularly in the group of children with idiopathic short stature, possibilities of molecular analysis are often underestimated. Important options are UPD7 and the FGFR3, SHOX, GH1 and GHR genes. Furthermore, analysis of the IGF and IGF1R genes should be considered. We propose a flow chart for molecular analysis in short stature. PMID- 14530603 TI - Acarbose and idiopathic reactive hypoglycemia. AB - BACKGROUND/AIM: Therapy for idiopathic reactive hypoglycemia is ineffective and impractical. Acarbose, an alpha-glucosidase inhibitor, decreases the absorbed glucose load. This study was done to determine the efficacy of acarbose in this syndrome. METHODS: Six patients with symptoms suggestive of postprandial hypoglycemia were chosen consecutively. Three-hour oral glucose tolerance tests using 75 g of glucose were done on each patient. The patients were then treated with acarbose with each meal. RESULTS: All patients had symptomatic hypoglycemia during testing. After 4 weeks of acarbose therapy, all patients were asymptomatic on a regular diet. CONCLUSION: Acarbose is efficacious in the management of idiopathic reactive hypoglycemia. PMID- 14530604 TI - Long-term follow-up of childhood-onset hypopituitarism in patients with the PROP 1 gene mutation. AB - OBJECTIVE: The PROP-1 gene mutation is a rare disorder leading to combined pituitary hormone deficiencies over time. The aim was to analyze the clinical picture of 40 years of an almost untreated PROP-1 gene mutation. METHODS: We describe the clinical and hormonal data of 2 brothers from childhood to adulthood as well as imaging procedures (MRI of the pituitary gland, bone mineral density by QCT and DPX). The PROP-1 gene mutation (301-302delAG) was confirmed by DNA sequencing. RESULTS: Although long-standing untreated hypopituitarism was present, there was normal physical and professional activity. Bone mineral density was low only in 1 patient. Adrenocortical deficiency occurred late at 45 and 39 years. CONCLUSIONS: The biological evolution of the PROP-1 gene mutation illustrates the importance of continuous care for these patients. Hormonal deficiencies do not necessarily lead to the same phenotype as is obvious in differences of bone age and bone mineral density. PMID- 14530605 TI - Pilot study of elevated levels of insulin-like growth factor-binding protein-2 as indicators of hepatocellular carcinoma. AB - BACKGROUND/AIMS: Insulin-like growth factor-binding protein-2 (IGFBP-2) is expressed in many malignant tissues, and elevated serum levels can be indicators of tumour activity in addition to conventional tumour markers. Our aim was to evaluate the role of IGFBP-2 levels together with insulin-like growth factor (IGF)-I, IGF-II and IGFBP-3 in the diagnostic work-up of patients with hepatocellular carcinoma (HCC). METHODS: In 50 (39 males, 11 females) histologically confirmed and TNM-graded patients with HCC who had not received adjuvant chemotherapy, the basal serum levels of IGF-I, IGF-II, IGFBP-3, IGFBP-2 and alpha-fetoprotein (AFP) were measured. The median age of the patients was 66 (37-84) years, body mass index was normal (25 (35-16) kg/m2). RESULTS: The levels of IGF-I, IGF-II and IGFBP-3 were diminished, as is the case when nutrition, hepatic function and growth hormone (GH) secretion are decreased. The levels of AFP and IGFBP-2 were markedly high. In 37 cases, IGFBP-2 levels were above the age-related norm, and in 40 cases AFP levels were also elevated. In 3 cases, both AFP and IGFBP-3 were normal, and in 4 cases AFP was high but IGFBP-2 normal, whereas in 10 cases AFP was normal but IGFBP-2 was high. CONCLUSIONS: In addition to AFP, IGFBP-2 appears to be a suitable marker for the evaluation of the serological status of HCC patients. A longitudinal study during disease management is required to assess the full potential of IGFBP-2 measurements as a marker. PMID- 14530606 TI - Growth velocity: a presentation of reference values in Dutch children. AB - Normal values are given for height velocity in a Dutch population. A construction of velocity curves is given and a comparison is made with other studies. PMID- 14530607 TI - Leptin in relation to hepatocellular carcinoma in patients with liver cirrhosis. AB - OBJECTIVES: This study was conducted to investigate whether leptin is involved in the etiogenesis of hepatocellular carcinoma (HCC) in cirrhotic patients. METHODS: Thirty-one male cirrhotic patients with HCC, 26 male cirrhotic patients without HCC, and 25 control subjects were included in this study. Body fat mass (FM) was determined by bioelectrical impedance analysis, and serum leptin and hormone concentrations were measured by immunoassay. RESULTS: A significant correlation of serum leptin levels to FM was observed in both patient groups and control subjects (r = 0.760, p < 0.001; r = 0.520, p < 0.01; r = 0.460, p < 0.05, respectively). The serum leptin levels in cirrhotic patients with or without HCC were significantly higher than those in control subjects (6.0 +/- 1.1 vs. 6.1 +/- 0.6 vs. 3.8 +/- 0.3 ng/ml, p < 0.05), though their body FM was lower. Using a multiple logistic regression analysis, it was found that the odds ratio of serum leptin for HCC was 1.04 (95% CI 0.79-1.33) after adjustment of several known risk factors. CONCLUSIONS: Our study demonstrated that cirrhotic patients with or without HCC had increased serum leptin concentrations. However, leptin did not appear to be associated with the development of HCC in cirrhotic patients. PMID- 14530608 TI - Significance of low-dose and standard-dose ACTH tests compared to overnight metyrapone test in the diagnosis of adrenal insufficiency in childhood. AB - OBJECTIVE: To discover the value of low-dose (LDAT) and standard-dose ACTH tests (SDAT) as compared with the metyrapone test in the diagnosis of secondary adrenal insufficiency. PATIENTS AND METHODS: LDAT (0.5 microg/m2), SDAT (250 microg/m2) and overnight metyrapone (30 mg/kg) tests were carried out in 29 patients with suspected adrenal insufficiency. LDAT and SDAT were also performed in 36 control subjects. RESULTS: 18 of 29 patients were grouped in the adrenal-sufficient (AS) group and 11 of 29 patients in the adrenal-deficient (AD) group according to the metyrapone test results. The control group had significantly higher cortisol responses than the AS and AD groups during LDAT. The control group had similar cortisol responses to the AS group but higher cortisol responses than the AD group during SDAT. The AS group was divided into 2 subgroups: AS patients with multiple pituitary hormone deficiencies (AS-multiple) and AS patients with idiopathic growth hormone deficiencies (AS-isolated). The AS-multiple group had statistically lower cortisol responses than the control group during LDAT. Receiver-operating characteristics analysis revealed that the cortisol cutoff value in LDAT was 19.8 microg/dl (100% sensitivity, 89% specificity) and 30.4 microg/dl in SDAT (82% sensitivity, 78% specificity). CONCLUSION: LDAT is capable of identifying patients with adrenal insufficiency more effectively than SDAT. The cortisol cutoff value in LDAT was calculated as 19.8 microg/dl with 100% sensitivity. AS patients with multiple pituitary hormone deficiencies had lower cortisol responses to LDAT than the control group implying that these patients might have a lower cortisol secretory capacity than healthy subjects. PMID- 14530609 TI - The uterine length in women with Turner syndrome reflects the postmenarcheal daily estrogen dose. AB - AIM: To evaluate the effects of estrogen substitution on the uterine development in patients with Turner syndrome. METHOD: 57 women, aged 18.1-41.5 years, were treated with estrogen from puberty induction. RESULTS: In 21 women (37%), the uterus developed to >65 mm in length. The daily estrogen dose correlated with both uterine length (r = 0.29; p < 0.05) and Tanner breast stage (r = 0.44; p < 0.001). A negative correlation between age at artificial menarche and uterine length was found (r = -0.29; p < 0.05). The endometrium thickness was greater in women with an uterus length >65 mm (p < 0.05). In 50% of the women (18 were evaluated), an adult-shaped uterus developed. Previous growth hormone therapy (n = 32) had no impact on the uterus length. CONCLUSIONS: The uterine development was suboptimal in most patients. Further investigation is needed to optimize estrogen therapy for uterine development in patients with Turner syndrome. PMID- 14530610 TI - Scintigraphy with [111In]octreotide and 201Tl in a Hurthle cell thyroid carcinoma without detectable radio-iodine uptake. Report of a case and review of the literature. AB - AIM: To describe the visualization of recurrent disease by [111In]octreotide and 201Tl scintigraphy in a patient with Hurthle cell thyroid carcinoma, increased thyroglobulin levels, and a negative radio-iodine total-body scan. METHODS: Scintigraphy with [111In]octreotide and 201Tl was performed, and a local recurrence in the thyroid bed was detected which was excised by surgery. RESULTS: On histology, the tumour proved to be a Hurthle cell carcinoma, and within the tumour somatostatin receptors were detected by RT-PCR. CONCLUSION: Scintigraphy with [111In]octreotide and 201Tl is an alternative imaging method for the detection of residual disease in patients with a differentiated thyroid carcinoma having increased thyroglobulin levels and a negative radio-iodine total-body scan. PMID- 14530611 TI - Family history of hypertension and diabetes in relation to preeclampsia risk in Peruvian women. AB - In a case-control study of 169 preeclamptics and 201 controls, we assessed maternal parental history of chronic hypertension and diabetes in relation to preeclampsia risk among Peruvian women. Participants provided information on parental history of the two conditions and other covariates during postpartum interviews. Logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs) adjusted for confounding by age, parity and prepregnancy adiposity. In this population, women were more likely to know the diabetes status of their parents than their hypertension status. Compared with women without a parental history of hypertension, women with a parental history of hypertension experienced a 20% increased risk of preeclampsia (OR = 1.2; 95% CI 0.7-2.2) that did not reach statistical significance. Women with a positive parental history for diabetes had a 3.4-fold increased risk of preeclampsia (95% CI 1.4-8.4). Women with a positive parental history of both hypertension and diabetes, as compared with those whose parents had neither condition, experienced a 4.6- fold increased risk of preeclampsia (OR = 4.6; 95% CI 0.9-23.0). Our results are generally consistent with the thesis that parental history of hypertension and diabetes reflects genetic and behavioral factors whereby women may be predisposed to an increased risk of preeclampsia. PMID- 14530612 TI - Intervention rates after elective induction of labor compared to labor with a spontaneous onset. A matched cohort study. AB - INTRODUCTION: Elective induction of labor has become a widely used procedure in obstetrics. A number of studies have shown an increased incidence of operative deliveries. The objective of this study was to evaluate the rate of interventions in our hospital, including operative delivery. METHODS: A matched cohort study in which labor of 122 electively induced women and 122 women with labor with a spontaneous onset were analyzed retrospectively. These women were matched for parity and gestational age. RESULTS: Pain relief, fetal scalp blood sampling and operative deliveries were recorded more frequently in the electively induced labor group. Cesarean delivery was found in 15% of women with induced labor, and in 1% of labors with a spontaneous onset (relative risk 18 (95% CI 2.4-132.7)). No differences were found in neonatal outcomes. CONCLUSIONS: Elective induction of labor leads to increased intervention rates during labor. The rate of cesarean delivery is high, particular in nulliparous women and multiparous women without a previous vaginal birth. PMID- 14530613 TI - Doppler velocimetry and cytofluorimetric analysis in uterine myomas. AB - BACKGROUND: To correlate resistance indices (RI) of vessels detected by transvaginal Doppler velocimetry to the proliferative fraction (PF) calculated by flow cytometry in uterine myomas. A prospective study was carried out on patients scheduled for surgery because of symptomatic uterine myomas. A group of 26 myomas characterized by the presence of detectable vessels at Doppler analysis in the inner part of the mass were included in the study. METHODS: The day before surgery patients were studied with both conventional and color Doppler ultrasonography. The volume of the myoma was recorded. The lowest value of RI obtained by Doppler analysis of the vessels within the myoma was considered for the study. Samples from myomas were processed and analyzed by flow cytometry. A PF, representing the cumulative frequency of cells in the S and G2/M phases of the cell cycle, was considered for the analysis. RESULTS: No statistically significant correlation was observed between RI and PF (r = 0.06, p = 0.74). Regression analysis of the intratumoral RI values on the volume of the myoma showed a significant negative correlation (r = -0.46, p = 0.012). CONCLUSIONS: Preoperative Doppler evaluation of uterine myomas did not provide predictive information of the proliferative status of the neoplasm. A significant correlation was found between the volume of the myoma and RI. PMID- 14530614 TI - The role of nitric oxide on apoptosis in human luteinized granulosa cells. Immunocytochemical evidence. AB - This study was designed to investigate the role of nitric oxide (NO) on the apoptosis of human luteinized granulosa cells and its possible pathways. Granulosa cell suspensions were incubated for 48 h after adding NO donor (S nitroso-N-acetyl-penicillamine, SNAP) and NO synthase inhibitor (nitro-L-arginine methyl ester, L-NAME) at different concentrations. Apoptosis was examined using a terminal deoxynucleotide-transferase-mediated dUTP-biotin nick end labeling method in 70 patients, and immunocytochemical staining was performed for six apoptosis-related proteins in 50 patients. Apoptotic rates were significantly lower in cells incubated with 0.5 mM SNAP, but higher with 0.5, 1.0, and 5.0 mM L NAME. SNAP (0.5 mM) lowered the expression of Fas and p53 in luteinized granulosa cells, but Bcl-2 expression was increased, and Fas ligand or Bax remained unchanged. Using L-NAME (0.5 and 5.0 mM), the expression of p53 and Bax was increased, but Bcl-2 was unchanged. Fas/Fas ligands were also activated especially in 5.0 mM L-NAME. In conclusion, NO may inhibit apoptosis via decreased Fas and p53, and increased Bcl-2 expression in human luteinized granulosa cells. PMID- 14530615 TI - Conservative management of ectopic pregnancy with fetal cardiac activity by combined local (sonographically guided) and systemic injection of methotrexate. AB - OBJECTIVE: To evaluate the efficacy of conservative management of ectopic pregnancy with fetal cardiac activity by combined local sonographically guided and systemic injection of methotrexate. STUDY DESIGN: The study group included 12 patients with ectopic pregnancy and fetal cardiac activity, treated by combined local and systemic injection of methotrexate in the period from January 1, 2000 to July 1, 2002. The outcome of these patients was compared with the outcome of 53 patients who had ectopic pregnancy without fetal cardiac activity and were being treated only by systemic injection of methotrexate during the same period of time. RESULTS: The success rate was 91.6% (11 out of 12) in the group of patients with ectopic fetal cardiac activity and 90.5% (48 out of 53) in the group of patients with ectopic pregnancy, but without fetal cardiac activity. There was also no significant difference between the two groups comparing the percentage of cases treated by an additional dose of methotrexate (8.3 and 13.2%, respectively), nor comparing the number of days to resolution of beta-human chorionic gonadotrophin (BHCG; 40 +/- 2 and 34 +/- 10 days, respectively). Conversely, there was a significant difference in the initial BHCG level comparing the group of patients treated by combined local and systemic injection of methotrexate (12,616 +/- 9,585 mIU/ml) and the group of patients treated by systemic injection of methotrexate (1,499 +/- 2,065 mIU/ml) (p < 0.00001). Seventy-five percent of patients (6 out of 8) diagnosed with ectopic fetal cardiac activity, who desired to become pregnant, succeeded to conceive within 6 months following the combined local and systemic injection of methotrexate. CONCLUSION: The combined local sonographically guided and systemic injection of methotrexate is associated with a successful outcome in asymptomatic patients presenting with ectopic pregnancy and fetal cardiac activity. PMID- 14530616 TI - 17beta-estradiol- and lipopolysaccharide-induced changes in nitric oxide, tumor necrosis factor-alpha and vascular endothelial growth factor release from RAW 264.7 macrophages. AB - The present study investigated the effect of 17beta-estradiol (17betaE(2)) and lipopolysaccharide (LPS) on the release and expression of nitric oxide (NO), tumor necrosis factor-alpha (TNF-alpha) and vascular endothelial growth factor (VEGF) in the macrophage cell line RAW 264.7. The cells were pre-incubated with 17betaE(2) at physiological as well as supraphysiological concentrations (12.5 1000 pg/ml) and subsequently activated with LPS (100 ng/ml). The changes in NO, TNF-alpha and VEGF release into culture medium and also their gene expression in cells were assessed. A concentration-dependent inhibitory effect of 17betaE(2) on NO and TNFalpha release was observed at low physiological concentrations, whereas the VEGF gene expression and protein levels were upregulated by 17betaE(2) in RAW 264.7 cells. PMID- 14530617 TI - Primary and secondary stroke prevention in nonrheumatic atrial fibrillation by oral anticoagulation. AB - The risk of stroke or embolism in atrial fibrillation (AF) patients can be reduced by 68% by oral anticoagulation (OAC). This review is aimed to (1) summarize indications for OAC in patients with AF, (2) give an overview of the current knowledge of risk factors for bleeding complications of OAC and (3) give practical recommendations for an optimal OAC therapy in the neurological setting. Indications for OAC are increased age (>75 years), arterial hypertension, diabetes mellitus, previous thromboembolism, heart failure and, probably, coronary heart disease. Risk factors for bleeding complications are overanticoagulation with international normalized ratios (INRs) >3.0, increased age, arterial hypertension, diabetes mellitus, previous thromboembolism, polypharmacy, the early phase of OAC therapy and a lack of patients' education. Before initiation of OAC, the patient should be screened for potential bleeding sites. Careful monitoring of OAC comprises fixed appointments for the INR value determination, tracking for the patient, if he does not attend, advices about pain therapy, information about the influence of diet on the INR value and drug interaction, unscheduled INR determination in case of acute disorders and regular assessment if OAC is still indicated. Monitoring of OAC needs an effort, which has to be adequately estimated by the health care system. PMID- 14530618 TI - Etiology and long-term prognosis of unilateral paramedian pontine infarction with progressive symptoms. AB - Progressive pure motor hemiparesis is a common feature in paramedian pontine infarction. To assess the etiology, clinical course and long-term prognosis of this stroke subtype, we identified 26 patients with progressive symptoms in a retrospective study, treated in our hospital between 1993 and 1998. All patients had more than one stroke risk factor, and in 20, basilar branch disease, an atherosclerotic occlusion of basilar perforating arteries, was causative. Most patients developed severe hemiparesis after a mean time of 3 days, nearly 50% showed deterioration in connection with a fall in systolic blood pressure. The patients were followed up for 4-9 years: 4 patients had further strokes, 5 died, and 17 patients were independent, which may be interpreted as a good long-term prognosis. PMID- 14530619 TI - Microembolus detection in four patients with Fabry's disease: further support for a primarily microangiopathic origin of early cerebrovascular symptoms. AB - Fabry's disease is a sex-linked recessive storage disorder leading to cerebrovascular symptoms in many cases. Whether the primary origin of cerebrovascular symptoms in these patients is due to autochthonous thrombosis of small vessels or embolic infarction from large vessels is under debate. Microembolic signals (MES) indicative of an active embolic source can be recorded by transcranial Doppler sonography. We report four patients (34, 44, 62 and 66 years old) with Fabry's disease from one kindred in whom we carried out microembolus detection recordings to disclose active embolic sources. Three of the patients had had transient focal neurological deficits. Two patients had angina pectoris and one had exercise dyspnea, but no patient had clear evidence of disease of the large coronary vessels or brain-supplying arteries. No MES were found in 1-hour recordings in any patients, supporting the hypothesis that early cerebrovascular symptoms arise from small vessel occlusion and not from embolic sources as an indicator of disease of large brain-supplying arteries or cardiac embolism. PMID- 14530620 TI - Neurophysiological diagnosis of acquired sensory ganglionopathies. AB - We examined 29 patients with chronic progressive ganglionopathy of different etiology. Neurophysiological abnormalities were dominated by a widespread decrease in sensory nerve action potential amplitudes, which involved both upper and lower limb nerves, even in patients with asymmetrical or patchy clinical presentation. This impairment of sensory nerve conduction, reflecting a nonlenght dependent pattern of peripheral axon degeneration, should be considered the hallmark of ganglionopathies. The evidence of central sensory pathway impairment, which allows to localize the pathology to the dorsal root ganglion neurons, could be better confirmed by cervical magnetic resonance imaging, which showed a diffuse hyperintensity in the posterior columns in all patients, than by somatosensory evoked potentials, which were undetectable in most of the patients. Few patients showed an impairment of individual motor nerves, which appeared more severe in paraneoplastic associated ganglionopathies. Neurophysiological abnormalities did not appear significantly changed at the 4-year follow-up. We emphasize that distinct abnormalities allow to differentiate ganglionopathies from axonal sensory neuropathies on routine neurophysiological examination. PMID- 14530621 TI - Movement disorders possibly induced by traditional chinese herbs. AB - The authors describe the neurological presentation and CT/MRI findings in 4 patients exposed to overdoses of decoctions of two different Chinese herbs. Case 1, a 15-year-old boy, ingested herba serissae along with the safe-dosage Salvia miltiorrhiza for treating a left renal stone. Sophora subprostrata root (SSR) was primarily used for treating three other diseases: viral B hepatitis in case 2, a 9-year-old boy; infection of the throat and a low fever in case 3, a 11-year-old girl, and a minor facial infection in case 4, a 12-year-old boy. All patients showed complex neurological manifestations primarily including convulsions, mental changes and dystonia syndromes. Their CT and/or MRI revealed abnormal density lesions in the striatum and globus pallidus bilaterally. They excluded the possibility of Wilson's disease in each of the 4 patients and suggested that overdosage of SSR and herba serissae could cause intoxications of the central nervous system, particularly damage to the basal ganglia. Chemically, coumarin (case 1) and matrine and oxymatrine (cases 2-4) in the two medicinal herbs are suggested to be possibly responsible for the morbidity. PMID- 14530622 TI - Assessment of driving performance in patients with relapsing-remitting multiple sclerosis by a driving simulator. AB - OBJECTIVE: To compare the driving performance using a driving simulator with physical and cognitive functions as measured by the Expanded Disability Status Scale (EDSS) and the Multiple Sclerosis Functional Composite (MSFC) in patients suffering from the relapsing-remitting form of multiple sclerosis (RRMS). METHODS: 31 RRMS patients (18 women, 13 men, mean age 35.6 +/- 8.3 years, EDSS 2.8 +/- 1.4) were compared with 10 healthy controls (8 men, 2 woman, age 45.1 +/- 7.8 years). RESULTS: Compared with controls, the accident rate (5.3 +/- 3.8 vs. 1.3 +/- 1.5, p < 0.001) and concentration faults (21.1 +/- 15.5 vs. 7.1 +/- 2.6, p < 0.01) of RRMS patients using the driving simulator were increased. While there was no correlation with the EDSS score, the accident rate was correlated with the MSFC (r = -0.5, p < 0.05). Regarding the three dimensions of the MSFC, accidents were related to the number of correct answers and Z-score in the paced auditory serial addition test (PASAT) as a measure for cognitive function (r = 0.33, p < 0.05). CONCLUSION: The current study demonstrates the need to focus also on driving skills in MS patients. The risk of accidents should be evaluated after relapses in particular. However, there are great interindividual differences. In the MSFC, most deficits could be evaluated in the PASAT. As there was a significant correlation between the accident rate in the driving simulator and the PASAT results, accidents seem to be more influenced by cognitive decline than by physical impairment. This indicates that the MSFC is a broader, more dimensional scale than the EDSS and should be preferred in the case of driving assessment. At the present time, the driving simulator seems to be a useful instrument judging driving ability, especially in cases with ambiguous neuropsychological results. PMID- 14530623 TI - Is the reduction of spasticity by botulinum toxin a beneficial for the recovery of motor function of arm and hand in stroke patients? AB - To determine the functional benefit of botulinum toxin A (BtxA) in spasticity of arm flexors, we conducted an open-label study with 10 stroke patients. 480 mouse units BtxA (Dysport((R))) were injected into flexor muscles. Outcome measures were done by the Modified Ashworth Scale, Rivermead Motor Assessment (arm section), active and passive ranges of motion (ROM), grip strength and a 3 dimensional motion analysis. The functional capacity of the entire arm, even of the shoulder, and the ROM of fingers and wrist increased. Despite a diminution of muscle tone, grip strength remained unchanged or increased. An improvement of forearm extension was only observed in cases of preserved voluntary extensor motricity. Injection of BtxA into spastic distal flexor muscles may lead to a functional improvement of the entire arm. Grip strength is not necessarily reduced. PMID- 14530624 TI - Therapy with intravenous immunoglobulins: complications and side-effects. AB - Therapy with intravenous immunoglobulins (IVIG) is thought to be a safe treatment for a number of immune-mediated neurological diseases. Published data about prevalence of adverse effects range from 11 to 81%. The purpose of our study was to present a representative view on adverse effects by analysis of a large cohort of patients treated by IVIG. In a prospective study, we analysed 117 patients (age 17-79 years) who were treated with IVIG for various neurological diseases including chronic inflammatory demyelinating polyneuropathy, diabetic amyotrophy, inclusion body myositis, multiple sclerosis, Guillain-Barre syndrome, Miller Fisher syndrome, multifocal motor neuropathy, myasthenia gravis and polymyositis. IVIG therapy was applied at a dose of 0.4 g/kg body weight/day in a total of 408 therapy courses. 42.7% showed adverse events. The majority of patients presented with minor adverse effects, mostly asymptomatic laboratory changes. Rash or mild headache occurred in 8 patients, especially when IVIG was given with infusion flow higher than 10 g/h. Two patients showed a severe complication with deep vein thrombosis. In summary, beside its effectiveness in immune mediated neurological diseases, therapy with IVIG seems to be a safe therapy. Most patients show no or minor adverse effects. Patients with pre-existent disorders like heart or renal insufficiency or immobilised patients, however, may be at higher risk for complications. PMID- 14530625 TI - Effect of sleep deprivation on saccades and eyelid blinking. AB - In this study the effect of sleep deprivation on specific components of eye and eyelid movement was investigated in a group of young and healthy subjects. The duration of sleep deprivation was 20 h. Each subject had to execute different saccade tasks: reflexive saccades, voluntary prosaccades and antisaccades. Saccade latency, number of saccade errors and blink rate during the saccade tasks were evaluated as predictors of performance decrements resulting from sleep deprivation. The present study showed no significant deterioration of latency and number of errors in the different saccade tasks. However, the blink rate was significantly higher after a night without sleep than before. It is concluded that the blink rate appears to be a promising psychophysical measurement and a more sensitive parameter than saccade performance to evaluate the effects of 20 h of sleep deprivation. These results encourage further research on this topic. PMID- 14530626 TI - Isolated trochlear nerve palsy in CREST syndrome. PMID- 14530627 TI - Schizophreniform disorder with cerebrospinal fluid PCR positivity for herpes simplex virus type 1. PMID- 14530628 TI - Acute axonal polyneuropathy associated with alcoholism. PMID- 14530629 TI - Wernicke encephalopathy - a severe neurological complication in a clinically inactive Crohn's disease. PMID- 14530630 TI - Dissociated preservation of automatic-voluntary jaw movements in a patient with biopercular and unilateral pontine infarcts. PMID- 14530631 TI - Diffusion-weighted MRI findings in patients with capsular warning syndrome. AB - BACKGROUND AND PURPOSE: The 'capsular warning syndrome' (CWS) of recurrent stereotyped episodes of motor or sensory dysfunction is clinically well recognized, and is associated with a high risk of imminent lacunar infarction with permanent deficits resembling those of CWS. However, the pathophysiology of CWS has not been well characterized. We report a clinicoanatomic correlation with MR imaging studies in the acute and chronic phases in patients with CWS. MATERIAL AND METHODS: Between April 1997 and March 2001, we prospectively studied 8 patients, mean age 73.3 years, presenting with 4-17 motor or sensorimotor transient ischemic attacks (TIAs; duration 2-90 min) up to 3 days after onset of the first episode. Four patients were free of symptoms between the attacks and had no residua, whereas 4 patients developed a pure motor or sensorimotor stroke within 1-3 days after symptom onset. Diffusion-weighted echoplanar MRI (DWI) and T(2)-weighted MRI studies were performed within 1 week after symptom onset and were repeated 1-2 months later. RESULTS: Seven of the 8 patients had an appropriate lesion on DWI in the acute phase. DWI abnormalities in the 3 patients with TIAs were 4-10 mm in diameter and confined to the lateral thalamus or medial globus pallidus without involving the internal capsule, whereas 4 patients who developed a stroke had abnormalities localized to the putamen extending to corona radiata (3 patients), or the pontomesencephalic junction (1 patient). All 6 patients who underwent follow-up MRI had an infarct on T(2)-weighted images corresponding to, but usually smaller than, the acute phase DWI abnormality. CONCLUSIONS: Small infarcts in the basal ganglia or the pons, close to central motor pathways, appear to be the primary lesion in CWS. The pathophysiology of CWS is complex, and may involve hemodynamic mechanisms in penetrating arterial territories, as well as molecular mechanisms, such as peri-infarct depolarizations affecting adjacent motor pathways. PMID- 14530632 TI - Cerebral sinus venous thrombosis in two patients with spontaneous intracranial hypotension. AB - We report 2 patients who had clinical and neuroimaging signs of spontaneous intracranial hypotension and who developed cerebral sinus venous thrombosis. This sequence of events -- known after dural puncture but not in spontaneous intracranial hypotension -- was suggested by the change in the pattern of headache, from a postural to a permanent and increasing one. The diagnostic and therapeutic difficulties that this complication raises are discussed. PMID- 14530633 TI - Early functional recovery and the fate of the diffusion/perfusion mismatch in patients with proximal middle cerebral artery occlusion. AB - BACKGROUND: The relationship between early neurological recovery, time to recanalization and the salvage of hypoperfused, but not diffusion-restricted tissue was investigated. METHODS: In 17 patients with acute middle cerebral artery occlusion, a multiparametric stroke MRI protocol was performed < 6 h after symptom onset, as well as at day 2 and 7. Recanalization was monitored with transcranial Doppler or with conventional angiography (during local thrombolysis). Functional improvement was defined as a change of > or = 4 points on the National Institutes of Health Stroke Scale score. RESULTS: In patients with functional improvement, 78% (median, range 66-95%) of the acute mean transit time (MTT) lesion escaped infarction compared with 28% (median, range -13 to 78%) in patients without neurological improvement (p < 0.01). Similarly, the percentage of tissue with a time-to-peak (TTP) delay of > or = 2 s not progressing to infarction was 80 and 4% in the groups with and without improvement, respectively (p < 0.01). Neurological improvement was more frequent in patients with early (< or = 3 h after presentation) recanalization, due to the salvage of larger areas of initially hypoperfused tissue. CONCLUSIONS: The salvage of hypoperfused tissue is a major factor influencing early neurological improvement. PMID- 14530634 TI - Characteristics of the National Institute of Health Stroke Scale: results from a population-based stroke cohort at baseline and after one year. AB - BACKGROUND: The National Institute of Health Stroke Scale (NIHSS) results at baseline and after 1 year have never before been accounted for within an unselected population-based stroke sample. Neither has it been shown which individual items in the scale are the most important ones for the outcome in terms of death or dependency after 1 year. METHODS: The subjects were all patients within a municipality who had their first-ever non-subarachnoidal stroke during 1 year (n = 377). Impairment was evaluated at baseline (within 24-48 h) and after 1 year with the 15-item version of the NIHSS. At the 1-year follow-up, the Modified Rankin Scale was used in order to determine which patients were dependent. Predictors of death and dependency were analysed in logistic regression models. The different NIHSS items, age and gender were used as independent variables. RESULTS: The median NIHSS score was 6 (interquartile range 3-12) at baseline and 1 (interquartile range 0-3) at the 1-year follow-up, when 33% of the patients had died. Of patients scoring less than 4 on baseline NIHSS, 75% were functionally independent after 1 year. Seventeen per cent were functionally dependent and 8% were dead. Independent predictors of death were: age, questions, commands, gaze, alertness and sensation. Independent predictors of dependency were: age, commands, alertness and motor leg. CONCLUSIONS: Baseline NIHSS predicts the outcome after 1 year at the group level. Age and any reduction of the level of consciousness on arrival were associated with bad outcome after 1 year. PMID- 14530635 TI - Complications following carotid angioplasty and carotid stenting in patients with symptomatic carotid artery disease. AB - BACKGROUND: We review a single centre's experience of the endovascular treatment of carotid artery disease, present the 30-day and 1-year complication rates and assess whether changes in technique are associated with a change in clinical outcome. METHODS: Patients who underwent carotid angioplasty with or without stenting for symptomatic > or = 70% carotid artery stenosis secondary to atherosclerosis were included. 333 procedures were performed, i.e. angioplasty alone (86), stent without cerebral protection (150) and stent with cerebral protection (97). RESULTS: At 30 days, the total major disabling stroke and all death rate was 3.0%. For angioplasty alone, this was 2.3%, for stent without cerebral protection 4.0% and for stent with cerebral protection 2.1%. If non stroke-related deaths were excluded, it was 2.3, 3.3 and 0%, respectively. After 30 days, the 1-year ipsilateral stroke rate was 0.8%. CONCLUSIONS: Carotid artery stenting, in particular with a cerebral protection device, is a safe alternative to carotid endarterectomy for the treatment of symptomatic high-grade carotid artery disease. PMID- 14530636 TI - Aphasia after stroke: type, severity and prognosis. The Copenhagen aphasia study. AB - AIM: To determine the types, severity and evolution of aphasia in unselected, acute stroke patients and evaluate potential predictors for language outcome 1 year after stroke. METHODS: 270 acute stroke patients with aphasia (203 with first-ever strokes) were included consecutively and prospectively from three hospitals in Copenhagen, Denmark, and assessed with the Western Aphasia Battery. The assessment was repeated 1 year after stroke. RESULTS: The frequencies of the different types of aphasia in acute first-ever stroke were: global 32%, Broca's 12%, isolation 2%, transcortical motor 2%, Wernicke's 16%, transcortical sensory 7%, conduction 5% and anomic 25%. These figures are not substantially different from what has been found in previous studies of more or less selected populations. The type of aphasia always changed to a less severe form during the first year. Nonfluent aphasia could evolve into fluent aphasia (e.g., global to Wernicke's and Broca's to anomic), whereas a fluent aphasia never evolved into a nonfluent aphasia. One year after stroke, the following frequencies were found: global 7%, Broca's 13%, isolation 0%, transcortical motor 1%, Wernicke's 5%, transcortical sensory 0%, conduction 6% and anomic 29%. The distribution of aphasia types in acute and chronic aphasia is, thus, quite different. The outcome for language function was predicted by initial severity of the aphasia and by the initial stroke severity (assessed by the Scandinavian Stroke Scale), but not by age, sex or type of aphasia. Thus, a scoring of general stroke severity helps to improve the accuracy of the prognosis for the language function. One year after stroke, fluent aphasics were older than nonfluent aphasics, whereas such a difference was not found in the acute phase. PMID- 14530638 TI - Does treatment modality of intracranial ruptured aneurysms influence the incidence of cerebral vasospasm and clinical outcome? AB - BACKGROUND: Cerebral vasospasm is the most common cause of morbidity and mortality after aneurysmal subarachnoid hemorrhage (SAH). This study is designed to determine whether the incidence of symptomatic vasospasm and the overall clinical outcome differ between patients treated with surgical clipping compared with endovascular obliteration of aneurysms. METHODS: In this prospective study, 98 patients with aneurysmal SAH were treated. Seventy-two patients underwent surgery and clipping and 26 had coil embolization. The incidence of symptomatic vasospasm, permanent neurologic deficit due to vasospasm and clinical outcome were analyzed. Patients with better clinical and radiological grades (World Federation of Neurological Surgeons grades I-III and Fisher grades I-III) were analyzed separately. RESULTS: Symptomatic vasospasm occurred in 22% of the patients; 25% in the surgical group and 15% in the endovascular group. Nine percent of the patients in the surgical group and 7% in the endovascular group suffered ischemic infarction with permanent neurological deficit. These differences did not reach statistical significance (p = 0.42). For patients with better clinical and radiological grades, no significant difference was found for the rate of symptomatic vasospasm; 23% in the surgical and 12% in the endovascular group (p = 0.49). The overall clinical outcome was comparable in both groups, with no difference in the likelihood of a Glasgow Outcome Scale score of 3 or less (15% in the surgical and 16% in the endovascular group; p = 0.87). The same results for outcome were obtained for the subgroup of patients with better clinical grades on admission. CONCLUSION: Symptomatic vasospasm and ischemic infarction rate seem comparable in both groups, even for patients with better clinical and radiological admission grades. There is no significant difference in the overall clinical outcome at the long-term follow-up between both groups. PMID- 14530637 TI - Quality of life after treatment of unruptured intracranial aneurysms by neurosurgical clipping or by embolisation with coils. A prospective, observational study. AB - BACKGROUND: Relatively high rates of complications occur after operation for unruptured intracranial aneurysms. Published data on endovascular treatment suggest lower rates of complications. We measured the impact of treatment of unruptured aneurysms by clipping or coiling on functional health, quality of life, and the level of anxiety and depression. METHODS: In three centres, we prospectively collected data on patients with an unruptured aneurysm who were treated by clipping or coiling. Treatment assignment was left to the discretion of the treating physicians. Before, 3 and 12 months after treatment, we used standardised questionnaires to assess functional health (Rankin Scale score), quality of life (SF-36, EuroQol), and the level of anxiety and depression (Hospital Anxiety and Depression Scale). RESULTS: Nineteen patients were treated by coiling and 32 by clipping. In the surgical group, 4 patients (12%) had a permanent complication; 36 of all 37 aneurysms (97%) were successfully clipped. Three months after operation, quality of life was worse than before operation; 12 months after operation, it had improved but had not completely returned to baseline levels. Scores for depression were higher than in the general population. In the endovascular group, no complications with permanent deficits occurred; 16 of 19 aneurysms (84%) were occluded by more than 90%. One patient died from rupture of the previously coiled aneurysm. In the others, quality of life after 3 months and after 1 year was similar to that before treatment. CONCLUSIONS: In the short term, operation of patients with an unruptured aneurysm has a considerable impact on functional health and quality of life. After 1 year, recovery occurs but it is incomplete. Coil embolisation does not affect functional health and quality of life. PMID- 14530639 TI - Image quality of the temporal bone window examined by transcranial Doppler sonography and correlation with postmortem computed tomography measurements. AB - BACKGROUND AND PURPOSE: The major limitation of native transcranial colour-coded duplex sonography (TCCS) in older stroke patients is the relatively frequent occurrence of an insufficient temporal window. Our goal was to investigate the relationship between the in vivo Doppler ultrasound image quality of the temporal bone, and computed tomography (CT)-determined thickness, density, and homogeneity of cadaver temporal bone. METHODS: Thirty-three moribund neurological patients who eventually died were examined by TCCS using the transtemporal approach. The sonographer categorized the quality of the TCCS image (excellent, intermediate, and poor). During autopsy, a rectangular sample of the temporal squama was removed, which corresponded to the area of the in vivo acoustic window. The thickness of the whole temporal bone, cortical, and cancellous (= diploe) bone as well as the density and homogeneity were determined by high-resolution CT. RESULTS: Thirty-seven temporal bones were obtained. The quality of the acoustic window was classified as excellent in 13, intermediate in 6 and poor in 18 cases. A significant correlation between the complete bone thickness, as well as between the absolute thickness of the diploe and the quality of the acoustic window was found: the thinner the bone/diploe, the better the colour Doppler signal. The thickness of the cortical plates and the homogeneity of the bones were identical in the three image quality categories. CONCLUSION: The transtemporal TCCS image quality depends mainly on the thickness of the cancellous component of the temporal bone. PMID- 14530640 TI - Cause is the main predictor of outcome in patients with pontine hemorrhage. AB - BACKGROUND: Pontine hemorrhages frequently lead to death or severe disability. Predictors of early mortality have been previously described but little is known about the factors influencing long-term disability. OBJECTIVE: Determine clinical predictors of long-term outcome in a large hospital series of patients with acute pontine hemorrhage. METHODS: Review of all patients with acute pontine hemorrhage admitted to a single tertiary center from 1990 to 2002 who underwent a diagnostic workup including brain magnetic resonance imaging (MRI). Long-term outcome was rated using the modified Rankin Scale (mRS) score at the time of the last follow up. Odds ratios (ORs) for predictive factors were calculated using univariate and multivariable logistic regression analysis. Level of significance was established at p < 0.01. RESULTS: We identified 44 consecutive patients with acute pontine hemorrhage. Twenty-four patients had primary hemorrhages (no cause other than hypertension). Arteriovenous or cavernous malformations were documented by brain MRI in 20 patients. Patients with primary hemorrhages had worse Glasgow Coma Scale sum scores (p = 0.01) and more unfavorable CT patterns (p = 0.005) at presentation. Eighteen percent of the patients died in the hospital. Mean follow up among survivors was 20 months (range 3-144 months). Twenty-two percent of the patients remained severely disabled (mRS greater than 2) at 3 months and at the time of the last follow-up. All deaths occurred in patients with primary pontine hemorrhage. Poor outcome (death or severe disability) was more frequent in patients with primary hemorrhages when compared with patients harboring cavernous malformations (62 versus 5%; p < 0.01). Primary hemorrhage was the only independent predictor of poor outcome in multivariable regression analysis (OR 12; p = 0.029). CONCLUSIONS: Cause is a main predictor of clinical and radiological severity at presentation and outcome in patients with pontine hemorrhage. The first episode of hemorrhage in patients with brainstem cavernous malformations is usually benign. Pontine hemorrhage associated with prior hypertension is much more devastating. PMID- 14530641 TI - Unruptured aneurysms with cranial nerve symptoms: efficacy of endosaccular Guglielmi detachable coil treatment. AB - OBJECTIVE: To evaluate the efficacy of endosaccular Guglielmi detachable coil (GDC) treatment of unruptured aneurysms causing cranial nerve (CN) symptoms. MATERIALS AND METHODS: Among a database of 218 patients whose aneurysms were treated using GDC, seven patients met the criteria for unruptured aneurysms presenting with symptoms and signs of CN palsy. Changes in CN symptoms before and after GDC treatment were reviewed. RESULTS: Aneurysms were located in the internal carotid-posterior communicating artery (n=3), the basilar bifurcation (n=1) and the cavernous internal carotid artery (n=3). CN symptoms included ptosis (n=6), mydriasis (n=2), and extraocular muscle (EOM) disorder (CN III: n=4; CN VI: n=3). Overall, improvement or resolution of CN symptoms after treatment was noted in five patients. CN symptoms in cases involving small ( 0.05). The reading time for the LCD session was not significantly different from that for the CRT session (37.12 and 41.46 minutes, respectively; p = 0.889). CONCLUSION: For detecting small solitary pulmonary nodules, an LCD monitor and a CRT monitor are comparable. PMID- 14530644 TI - Differential diagnosis of benign and malignant intraductal papillary mucinous tumors of the pancreas: MR cholangiopancreatography and MR angiography. AB - OBJECTIVE: To compare the usefulness of magnetic resonance cholangiopancreatography (MRCP) and MR angiography (MRA) in differentiating malignant from benign intraductal papillary mucinous tumors of the pancreas (IPMTs), and to determine the findings which suggest malignancy. MATERIALS AND METHODS: During a 6-year period, 46 patients with IPMT underwent MRCP. Morphologically, tumor type was classified as main duct, branch duct, or combined. The diameter of the main pancreatic duct (MPD), the extent of the dilated MPD, and the location and size of the cystic lesion, septum, and communicating channel were assessed. For all types of IPMTs, enhanced mural nodules and portal vein narrowing were evaluated at MRA. RESULTS: Combined-type IPMTs were more frequently malignant (78%) than benign (42%) (p < 0.05). Compared with benign lesions, malignant lesions were larger, and the caliber of the communicating channel was also larger (p < 0.05). Their dilated MPD was more extensive and of greater diameter (p < 0.05), and the presence of mural nodules was more frequent (p < 0.001). CONCLUSION: Combined MRCP and MRA might be useful for the differential diagnosis of malignant and benign IPMTs of the pancreas. PMID- 14530645 TI - A comparative experimental study of the in-vitro efficiency of hypertonic saline enhanced hepatic bipolar and monopolar radiofrequency ablation. AB - OBJECTIVE: To compare the in-vitro efficiency of a hypertonic saline (HS) enhanced bipolar radiofrequency (RF) system with monopolar RF applications by assessing the temperature profile and dimensions of RF-created coagulation necrosis in bovine liver. MATERIALS AND METHODS: A total of 27 ablations were performed in explanted bovine livers. After placement of two 16-gauge open perfused electrodes at an interelectrode distance of 3 cm, 5% HS was instilled into tissue at a rate of 1 mL/min through the electrode. Seventeen thermal ablation zones were created in the monopolar mode (groups A, B), and ten more were created using the two open-perfused electrodes in the bipolar mode (group C). RF was applied to each electrode for 5 mins (for a total of 10 mins, group A) or 10 mins (for a total of 20 mins, group B) at 50W in the sequential monopolar mode, or to both electrodes for 10 min in the bipolar mode (group C). During RF instillation, we measured tissue temperature at the midpoint between the two electrodes. The dimensions of the thermal ablation zones and changes in impedance and wattage during RFA were compared between the groups. RESULTS: With open perfusion electrodes, the mean accumulated energy output value was lower in the bipolar mode (group C: 26675+/-3047 Watt s) than in the monopolar mode (group A: 28778+/-1300 Watt s) but the difference was not statistically significant (p > 0.05). In the bipolar mode, there were impedance rises of more than 700 ohm during RF energy application, but in the monopolar modes, impedance did not changed markedly. In the bipolar mode, however, the temperature at the mid-point between the two probes was higher (85 degrees C) than in the monopolar modes (65 degrees C, 80 degrees C for group A, B, respectively) (p < 0.05). In addition, in HS-enhanced bipolar RFA (group C), the shortest diameter at the midpoint between the two electrodes was greater than in either of the monopolar modes: 5.4+/-5.6 mm (group A); 28.8+/-8.2 mm (group B); 31.2+/-7.6 mm (group C) (p < 0.05) CONCLUSION: Using an open perfusion system, HS-enhanced bipolar RFA more efficiently created larger areas of thermal ablation and higher tissue temperatures than monopolar RFA. PMID- 14530646 TI - MR evaluation of radiation synovectomy of the knee by means of intra-articular injection of holmium-166-chitosan complex in patients with rheumatoid arthritis: results at 4-month follow-up. AB - OBJECTIVE: To determine whether MRI is able to demonstrate the effect of radiation synovectomy after the intra-articular injection of holmium-166-chitosan complex for the treatment of rheumatoid arthritis of the knee. MATERIALS AND METHODS: Fourteen patients aged 36-59 years were treated with 10-20 mCi of holmium-166-chitosan complex. A criterion for inclusion in this study was the absence of observable improvement after 3- or more months of treatment of the knee with disease-modifying anti-rheumatic drugs. MR images were acquired both prior to and 4-months after treatment. Clinical evaluation included the use of visual analog scales to assess pain, and the circumference of the knee and its range of motion were also determined. MR evaluation included measurement of the volume of synovial enhancement and wall thickness, the amount of joint effusion, and quantifiable scoring of bone erosion, bone edema and lymph nodes. RESULTS: Visual analog scale readings decreased significantly after radiation synovectomy (p < 0.05). MRI showed that joint effusion decreased significantly (p < 0.05), and that the volume of synovial enhancement tended to decrease, but to an insignificant extent (p = 0.107). CONCLUSION: The decreased joint effusion noted at 4-month follow-up resulted from radiation synovectomy of the rheumatoid knee by means of intra-articular injection of holmium-166-chitosan complex. PMID- 14530647 TI - Internal thoracic artery collateral to the external iliac artery in chronic aortoiliac occlusive disease. AB - OBJECTIVE: To evaluate the incidence and angiographic findings of the collateral pathway involving the internal thoracic artery in patients with chronic aortoiliac occlusive disease. MATERIALS AND METHODS: Between March 2000 and Februrary 2001, 124 patients at our hospital underwent angiographic evaluation of chronic aortoiliac occlusive disease, and in 15 of these complete obstruction or severe stenosis of the aortoiliac artery was identified. The aortograms and collateral arteriograms obtained, including internal thoracic arteriograms, as well as the medical records of the patients involved, were evaluated. RESULTS: In nine patients there was complete occlusion of the infrarenal aorta, or diffuse stenosis of 75% or more in the descending thoracic aorta, and in the other six, a patent aorta but complete occlusion or stenosis of 75% or more of the common iliac artery was demonstrated. Collateral perfusion via hypertrophied internal thoracic arteries and rich anastomoses between the superior and inferior epigastric arteries, reconstituting the external iliac artery, were noted in all fifteen patients, regardless of symptom duration, which ranged from six months to twelve years. CONCLUSION: In patients with chronic aortoiliac occlusive disease, the internal thoracic artery, along with visceral collaterals and those from the contralateral side, is one of the major parietal collateral pathways. PMID- 14530648 TI - Ultrasound evaluation of normal and abnormal fetuses: comparison of conventional, tissue harmonic, and pulse-inversion harmonic imaging techniques. AB - OBJECTIVE: To determine the usefulness of tissue harmonic imaging (THI) and pulse inversion harmonic imaging (PIHI) in the evaluation of normal and abnormal fetuses. MATERIALS AND METHODS: Forty-one pregnant women who bore a total of 31 normal and ten abnormal fetuses underwent conventional ultrasonography (CUS), and then THI and PIHI. US images of six organ systems, namely the brain, spine, heart, abdomen, extremities and face were compared between the three techniques in terms of overall conspicuity and the definition of borders and internal structures. RESULTS: For the brain, heart, abdomen and face, overall conspicuity at THI and PIHI was significantly better than at CUS (p < 0.05). There was, though, no significant difference between THI and PIHI. Affected organs in abnormal fetuses were more clearly depicted at THI and PIHI than at CUS. CONCLUSION: Both THI and PIHI appear to be superior to CUS for the evaluation of normal or abnormal structures, particularly the brain, heart, abdomen and face. PMID- 14530649 TI - Gossypiboma of the leg: MR imaging characteristics. A case report. AB - We report a 22-year-old man with a solid mass in the right proximal leg, which was furned out to be a gossypiboma. MR imaging revealed a well-defined mass lesion that showed intermediate signal intensity at T1-weighted imaging (T1WI) and slightly high signal intensity at T2-weighted imaging (T2WI). Wavy, low signal-intensity stripes were visible within the fluid-filled central cavity. At surgical exploration, a sponge, retained after previous knee surgery, was discovered, and it was found that a granuloma had developed. Pathologic examination revealed granulomatous inflammation, with lymphocyte and giant cell infiltration. The presence of wavy, low-signal-intensity gauze fibers at T2WI may be a characteristic MR appearance of gossypiboma. PMID- 14530650 TI - Sclerosing stromal tumor of the ovary: MR-pathologic correlation in three cases. AB - Sclerosing stromal tumor (SST) of the ovary is a very rare sex cord stromal tumor occurring in a younger age group than other types of stromal tumors and most commonly accompanied by menstrual irregularity. Several unique histologic features including pseudolobulation, sclerosis and prominent vascularity are clearly reflected at ultrasonography and MRI. We report the ultrasonographic and MR features of three cases of histologically confirmed SSTs, and relate them to the pathological findings. PMID- 14530652 TI - Addition of leukocytapheresis to steroid therapy: is it beneficial in recurrence of moderate-to-severe ulcerative colitis? AB - PURPOSE: In Japan, leukocytapheresis has become an accepted therapy for ulcerative colitis. The purpose of this retrospective study was to determine the efficacy of additional leukocytapheresis in patients with moderate-to-severe recurrence of ulcerative colitis. METHODS: From 1998 to 2002, 35 patients with moderate-to-severe recurrence of ulcerative colitis were treated by intravenous prednisolone only or prednisolone plus leukocytapheresis once per week. Previous clinical features of ulcerative colitis, activity index at four weeks, and subsequent course were compared between patients with leukocytapheresis and those without. RESULTS: Sixteen patients were treated with prednisolone only (prednisolone group), and 19 patients were treated with prednisolone plus leukocytapheresis (leukocytapheresis group). Previous prednisolone dose (13.6 +/- 10.4 g vs. 7.9 +/- 9.8 g; P < 0.05) was higher in the leukocytapheresis group than the prednisolone group. Clinical response rate at four weeks was not different between the two groups (leukocytapheresis group, 68.4 percent vs. prednisolone group, 75 percent). However, ulcerative colitis activity index at four weeks was significantly higher in the leukocytapheresis group than the prednisolone group (180.5 +/- 44.2 vs. 142.5 +/- 45.3; P < 0.05). During the subsequent clinical course, 15 of 35 patients required proctocolectomy (leukocytapheresis group, 11 patients vs. prednisolone group, 4 patients). Proctocolectomy rate was significantly different when patients were divided by previous prednisolone dose (P = 0.0029) and ulcerative colitis activity index at four weeks (P = 0.002), but it was not different according to the application of leukocytapheresis. Cox proportional hazards model revealed previous prednisolone dose to be the only independent risk factor for proctocolectomy. CONCLUSIONS: Addition of leukocytapheresis to prednisolone therapy does not seem beneficial in ulcerative colitis patients with moderate-to-severe recurrence. PMID- 14530653 TI - Hla-B genotype in Japanese patients with Crohn's disease. AB - PURPOSE: The HLA-B gene is one of the susceptibility genes for inflammatory bowel disease. Previous association studies of HLA-B showed several associated alleles and haplotypes of HLA-B in patients with ulcerative colitis, and among the associated alleles HLA-B*52 is well known to be strongly associated with ulcerative colitis in Japanese patients. However, there are no convincing reports about HLA-B including the B*52 allele in patients with Crohn's disease. The purpose of this study was to determine if HLA-B, especially the B*52 allele, confers susceptibility to Crohn's disease or determines the disease phenotype of Crohn's disease. METHODS: A total of 195 patients with Crohn's disease (49 ileitis, 106 ileocolitis, 34 colitis, 6 uncertain) and 185 healthy controls were studied in this case-controlled study. All patients and healthy controls were Japanese. Genotyping of the HLA-B gene was performed by a polymerase chain reaction, sequence-specific primer that can classify the gene into 23 allele groups. Allele frequencies were compared between patients with Crohn's disease and healthy controls with chi-squared test using a 2 x 2 contingency table. P value was corrected by the number of allele groups (n = 23) observed in the Japanese population or the number of clinical subgroups. Corrected P values of <0.05 were considered to be statistically significant. RESULTS: Before the correction for multiple testing, B*4001 and B*44 were associated with patients with Crohn's disease, positively and negatively, respectively. However, after the correction there were no significant differences in any HLA-B alleles between patients with Crohn's disease and healthy controls. In the subgroup analysis according to clinical phenotypes (disease location, anal lesion, age at diagnosis, need for surgery), none of the HLA-B alleles except B*52 showed any disease phenotype-genotype associations. The allele frequency of B*52 in the colitis type (16.2 percent; corrected P = 0.011) was significantly higher than that in the combined group of the ileitis (7.1 percent) and ileocolitis (5.2 percent) types. CONCLUSIONS: These results demonstrated that HLA-B did not confer overall susceptibility to Crohn's disease in Japan, but the B*52 allele may affect the location of the disease. PMID- 14530654 TI - Mortality and cause of death in Japanese patients with Crohn's disease. AB - PURPOSE: There are no reports based on long-term observation on mortality caused by Crohn's disease in Japan because of the small number of patients recorded throughout the 1970s. Because Crohn's disease patients in Japan are exempt from medical expenses, Crohn's disease is treated mainly with nutritional therapy and less frequently with corticosteroids. This study sought to estimate mortality and cause of death by Crohn's disease in Japan in the referred population. METHODS: Consecutive patients with Crohn's disease from 1967 to 1997 were identified in the Kyushu and Fukuoka University Study Group's nine affiliated hospitals and were followed up for 8.4 +/- 5.6 years. RESULTS: Death occurred in 6 of 544 patients. Cumulative survival rates for patients with Crohn's disease were 99.7 percent at 5 years, 99.3 percent at 10 years, and 96.8 percent at 15 years; there was no significant difference between genders or the age-matched expected survival rate of the Japanese population (P = 0.61, 0.84, 0.56). The overall standardized mortality rate was 1.43 (95 percent confidence interval, 0.53-3.12). The standardized mortality rate in newly diagnosed patients who were treated mainly with nutritional therapy was 0.75 (95 percent confidence interval, 0.02 4.18), and 1.69 in referred patients (95 percent confidence interval, 0.55-3.93). The standardized mortality rate by causes of death was high (64.4; 95 percent confidence interval, 7.72-232.5) in rectal cancer. In 150 patients who had been followed up for more than ten years, only 40 percent were administered corticosteroids. CONCLUSION: The standardized mortality rate of patients with Crohn's disease in the Kyushu and Fukuoka University Study Group's nine hospitals (the reference unit) was similar to reported rates of cohort studies. The standardized mortality rate of newly diagnosed patients was better than that of referred patients. The better vital prognosis of newly diagnosed patients might be attributable to the influence of frequent nutritional therapy and to lesser steroid use. PMID- 14530655 TI - Therapeutic results for hepatic metastasis of colorectal cancer with special reference to effectiveness of hepatectomy: analysis of prognostic factors for 763 cases recorded at 18 institutions. AB - PURPOSE: Factors affecting treatment prognosis and therapeutic results for hepatic metastasis of colorectal cancer were investigated. METHODS: Therapeutic results, especially of hepatectomy, were investigated for hepatic metastasis of colorectal cancer in 763 patients (585 underwent hepatectomy) treated between 1992 and 1996 at 18 institutions that participated in the "Study for establishing treatments for hepatic and pulmonary metastasis of colorectal cancer" sponsored by a Grant-in-Aid (10-11) for Cancer Research from the Ministry of Health, Welfare and Labor of Japan. RESULTS: The five-year survival rate for those treated by hepatectomy was significantly higher (32.9 percent) than for those not undergoing hepatectomy (3.4 percent). After hepatectomy for hepatic metastasis, the most prevalent form of recurrence was in the remnant liver (41.4 percent), followed by recurrence of pulmonary metastasis (19.2 percent), and other (7.2 percent). Factors of the primary tumor adversely affecting prognosis after hepatectomy for hepatic metastasis included poorly differentiated adenocarcinoma or mucinous carcinoma, depth of invasion of si/ai, lymph-node metastasis of Stage n3 and n4 by the Japanese classification of colorectal carcinoma, number of metastatic lymph nodes of more than four, and Dukes Stage D. Factors at the time of hepatectomy adversely affecting prognosis after surgery for hepatic metastasis included residual tumor, extrahepatic metastasis, hepatic metastasis of degree H3 stipulated by the Japanese classification of colorectal carcinoma, number of metastases of four or more, pathology of hepatic metastasis of poorly differentiated adenocarcinoma, resection margin of <10 mm, and carcinoembryonic antigen value higher than normal preoperative and one month postoperative. CONCLUSIONS: Among therapies for hepatic metastasis of colorectal cancer, the present study clearly revealed that hepatectomy is the treatment of choice whenever feasible. Postoperative recurrence often is localized in the remnant liver, or there may be a systemic recurrence of pulmonary metastasis. Thus, methods of prevention will be a future theme. PMID- 14530657 TI - Long-term prognosis of preoperative "bridge to surgery" expandable metallic stent insertion for obstructive colorectal cancer: comparison with emergency operation. AB - PURPOSE: Expandable metallic stent endoprosthesis for preoperative "bridge to surgery" treatment of obstructive colorectal cancer has been broadly, clinically used with good short-term results. Stent insertion for malignant tumor, however, is controversial because of the increased risk of metastasis, invasion, and advancement of the cancer. We, first in the world, evaluated the long-term prognosis of expandable metallic stent insertion compared with emergency surgery without expandable metallic stent. METHODS: Forty emergency operations (1986 1996) and 44 expandable metallic stent insertions (1993-2001) were retrospectively compared by history, duration, morbidity, and cumulative survival rate. RESULTS: There was no significant difference in age or lesion between the two groups. Postoperative complications were significantly less frequent in the expandable metallic stent group: wound infection was 14 vs. 2 percent; leakage following to anastomosis was 11 vs. 3 percent. Long-term prognosis did not significantly differ: three-year overall survival rate was 50 vs. 48 percent; five-year survival rate was 44 vs. 40 percent in the emergency operation and expandable metallic stent groups, respectively. CONCLUSION: Because preoperative expandable metallic stent insertion for obstructive colorectal cancer had good postoperative results and no disadvantages in long-term prognosis, this procedure should be used in preoperative treatments of obstructive colorectal cancer. PMID- 14530656 TI - Changes in colorectal cancer during a 20-year period: an extended report from the multi-institutional registry of large bowel cancer, Japan. AB - PURPOSE: This study was designed to examine trends of colorectal cancer in relation to age, gender, site, and survival during the past 20 years. METHODS: The multi-institutional registry of the Japanese Society for Cancer of the Colon and Rectum offered 87,695 surgical cases with invasive adenocarcinoma during 1978 to 1997 for analysis. We calculated survival rates and used the Cox's proportional hazard model for cases during 1978 to 1994. RESULTS: The number of cases showed a 2.5-fold increase with consistent male predominance confined to the distal colon and the rectum. Colon cancer in the last five-year period was more likely right-sided for females (odds ratio, 1.26; 95 percent confidence interval, 1.16-1.38) and males (odds ratio, 1.16; 95 percent confidence interval, 1.06-1.25) compared with the first period. Cancers in younger patients were more likely at Stage III to IV in the late 1990s if the cancers were in the distal colon, the rectum (for both genders), or the proximal colon (for females). Survival was improved except for cases with proximal colon cancer of Stage IV. In the multivariate analysis, hazard ratios for death in the postoperative five years were 0.77, 0.59, and 0.66 for proximal colon, distal colon, and rectal cancers, respectively, in the last period as compared with those in the first period [corrected]. Reduced hazard ratio for females was the largest for proximal colon cancer with Stage I to II. CONCLUSION: Although surgical outcome was largely improved, delayed presentation or diagnosis in younger patients remained a problem. Preferential localization in the proximal colon and survival benefit for females should be investigated. PMID- 14530658 TI - Results of linearly polarized near-infrared irradiation therapy in patients with intractable anorectal pain. AB - PURPOSE: Electrogalvanic stimulation and biofeedback therapy for the treatment of intractable anorectal pain have been reported. However, these therapeutic modalities have some disadvantages and insufficient effectiveness. We noticed that digital examination revealed the strongly tender point in both lateral sides of the rectum and introduced linearly polarized near-infrared irradiation therapy to the strongly tender point. The purpose of this study was to review the outcomes and estimate its usefulness. METHODS: A total of 35 consecutive patients complained of vague and deep pain in the anorectum. Fourteen patients had a history of lower abdominal surgery. Eighteen patients had disordered defecation. The linearly polarized near-infrared light was irradiated to the strongly tender point on or a few centimeters apart from the skin for ten minutes. The effect of the therapy was assessed as excellent, good, no change, or worse by the patients themselves. RESULTS: Ten patients had the strongly tender point in the left side, 8 in the right posterior, and 17 in both. Five patients estimated as excellent, 28 as good, and 2 as no change. Mean total number of irradiation was 18.8 (range, 1-235), and mean number of irradiation for relief from pain was 2.5 (range, 1-9). Anorectal pain recurred in four patients, who received the same therapy and improved. Four patients felt hot during the irradiation, and a patient had frequent micturition after the irradiation. These mild complications easily disappeared. CONCLUSION: The linearly polarized near-infrared irradiation therapy is a simple, safe, and effective modality for relief from intractable anorectal pain and recommended for primary therapy. PMID- 14530659 TI - Results of biofeedback therapy for fecal incontinence in children with encopresis and following surgery for anorectal malformations. AB - INTRODUCTION: Some children with fecal incontinence respond to biofeedback therapy. However, whether they can achieve fecal continence posttherapeutically has not been clarified. We studied the serial results of biofeedback therapy and discuss the necessity of providing repeated biofeedback therapy at home. METHODS: Nineteen children with encopresis underwent one session of biofeedback therapy. Seven of 15 children with fecal incontinence that developed after surgery for anorectal malformations underwent three to eight sessions of biofeedback therapy; the remaining 8 underwent one (mean, 2.9) session only. The patients were hospitalized for one session of biofeedback therapy. To monitor the clinical outcome of intervention, we used serial score assessments from three months to two years posttherapeutically. RESULTS: Seventeen of 19 (90 percent) patients with encopresis showed clinical improvement after one session of therapy (P < 0.0001). Six months after treatment, however, six of ten (60 percent) patients with encopresis reported recurrent fecal incontinence after one therapeutic session. Clinical improvement was noted in 5 of 15 (33 percent) patients who had fecal incontinence after surgery for anorectal malformations. All five patients showed clinical improvement from six months to two years after several sessions of biofeedback therapy (P < 0.05). CONCLUSIONS: Biofeedback therapy is effective in most children with encopresis and in some children with anorectal malformations. However, some patients need repeated sessions of biofeedback therapy to achieve fecal continence. Therefore, a new portable biofeedback apparatus for the treatment of fecal incontinence at home may be helpful. PMID- 14530660 TI - Intractable ulcerative colitis caused by cytomegalovirus infection: a prospective study on prevalence, diagnosis, and treatment. AB - PURPOSE: Cytomegalovirus infection has been known to complicate ulcerative colitis. This study was designed to elucidate the prevalence and clinical features of ulcerative colitis that might point efficiently to the diagnosis of complicating cytomegalovirus infection in cases of ulcerative colitis. METHODS: The study included 47 consecutive patients diagnosed to have moderate-to-severe ulcerative colitis who were treated on an inpatient basis at our department during a two-year period. A prospective examination for cytomegalovirus antigenemia was conducted in all patients with moderate-to-severe ulcerative colitis to determine the prevalence of cytomegalovirus infection among these patients. Then, the characteristic clinical and endoscopic features of ulcerative colitis complicated by cytomegalovirus infection were investigated by comparison of the cytomegalovirus-infected group with the non-cytomegalovirus-infected group. The therapeutic effects of antiviral drugs also were assessed. RESULTS: Cytomegalovirus infection was detected in 16 of 47 patients (34 percent). Proportion of female patients, age at the time of determination, and proportion of patients showing corticosteroid resistance was significantly higher in the cytomegalovirus-infected group (59.1 percent) than in the non-cytomegalovirus infected group (13.6 percent). The prevalence of endoscopically severe ulcerative colitis was higher in patients with cytomegalovirus antigenemia than in those without cytomegalovirus antigenemia (P = 0.016). Ganciclovir was administered to 12 of 16 ulcerative colitis patients with complicating cytomegalovirus infection, and was found to be effective in 8 (66.7 percent). CONCLUSIONS: It is not easy to make a diagnosis of cytomegalovirus infection complicating ulcerative colitis based on clinical features, including endoscopic biopsy. On the other hand, blood examination for the detection of cytomegalovirus antigenemia in corticosteroid resistant patients, particularly in relatively elderly patients, may enable diagnosis of cytomegalovirus infection with a high sensitivity and allow effective treatment to be administered in these patients. PMID- 14530661 TI - Leukocytapheresis as an adjunct to conventional medication for inflammatory bowel disease. AB - Inflammatory bowel disease is characterized by clinical remission and relapse caused by severe intestinal inflammation. Drug therapy for inflammatory bowel disease is associated with unpleasant side effects. Furthermore, efficacy of conventional drugs decreases with chronic use, which can be a major difficulty in the long-term management of this disease. In active inflammatory bowel disease, leukocytes are elevated with activation behavior and increased survival time, and mucosal neutrophil level parallels the severity of intestinal inflammation and predicts relapse. Leukocyte-derived inflammatory cytokines are suspected to be major factors in the initiation and perpetuation of inflammatory bowel disease. Accordingly, leukocytes should be appropriate targets for therapy. To reduce peripheral blood level of leukocytes, centrifugation has been used to deplete peripheral blood leukocytes; this provided the initial evidence that reducing the level of peripheral blood leukocytes can benefit patients with inflammatory disease. To overcome the limitations of centrifugation, membrane filters, such as the Cellsorba trade mark column and leukocyte-adsorbing beads containing column like Adacolumn, have been developed that are direct blood perfusion systems for removing any desired level of leukocytes. In initial independent clinical studies, these two new models have produced striking clinical efficacy, safety, and a marked reduction in the dose of corticosteroids used to induce remission of active inflammatory bowel disease. Leukocytapheresis has been associated with a significant decrease in the amount of several proinflammatory cytokines produced by peripheral blood leukocytes. Accordingly, the Japan Ministry of Health has now approved both methods for the treatment of active ulcerative colitis. Clinical data suggest that leukocytapheresis might be an effective adjunct to therapy for inflammatory bowel disease to promote remission, taper conventional drug dosage, and potentially reduce the number of patients who require colectomy. The results should further understanding of the pathophysiology of inflammatory bowel disease. PMID- 14530662 TI - Current status of autonomic nerve-preserving surgery for mid and lower rectal cancers: Japanese experience with lateral node dissection. AB - Surgical practices for treatment of rectal cancer in Japan have changed from extended dissection along perivascular or parietal plane to pelvic autonomic nerve-preserving procedures without compromising radicality of surgical resection. Previous surgical results suggested the significant advantages of extended surgery in survival and local recurrence rate of Dukes B and C patients. More than 15 percent of patients with cancer in the lower rectum have extramesorectal spread to lateral pelvic nodes that can be removed by lateral dissection for local control and cure. Initially the total nerve-preserving procedure has been introduced for a complete preservation of para-aortic and intrapelvic nervous system in patients with early-stage cancer not requiring para aortic and lateral lymph-node dissection. However, the concept of aggressive surgery for advanced rectal cancer has led to various types of pelvic autonomic nerve-preserving procedures, in which extended lymph-node dissection plus nerve preserving technique with resection of one or more autonomic nervous segments has been performed. During two decades, total pelvic autonomic nerve-preserving procedure with lateral lymph-node dissection has been used increasingly for Dukes C lesion without increased local recurrence. The overall status of pelvic autonomic nerve-preserving procedures according to clinical experiences in Japan is reviewed in the context of cadaveric anatomic findings, Japanese vs. Western techniques and concepts, and our own clinical data. PMID- 14530664 TI - Recent advances in diagnosis and treatment of colorectal T1 carcinoma. AB - PURPOSE: Colorectal T1 tumors (submucosally invasive carcinoma) have special characteristics in terms of pathology, diagnosis, and treatment. The clinicopathologic features of T1 tumors are reviewed. RESULTS: Incidence of T1 tumors was approximately 10 percent, and the percentage of node metastasis was 10 to 11 percent, whereas hepatic metastasis was <1 percent. Macroscopically they were divided into three types: polypoid, ulcerating, and flat. The ulcerating type was divided into two subgroups: polypoid growth and nonpolypoid growth based on the histologic appearance of resected specimens. The tools for detecting T1 tumors were fecal occult blood test and colonoscopy, and magnifying colonoscopy and chromography facilitate more precise diagnosis. Ultrasonography also was useful for the correct diagnosis of the depth of cancer invasion. Most polypoid and some flat T1 tumors were safely treated by polypectomy or endoscopic mucosal resection alone; however, when resected specimens contained risk factors for metastasis, such as deep invasion (sm2, sm3) and vessel invasion, additional surgery was necessary for cure. For rectal T1 tumors, the most appropriate procedure should be carefully selected from several therapeutic options to preserve anal function. CONCLUSION: The management of colorectal T1 tumors should be determined according to the types of macroscopic appearances. The risk of node metastasis could be predicted based on risk factors that were characterized by the level of cancer invasion and the presence or absence of vessel invasion. Minimally invasive treatment should be chosen for colorectal T1 tumors. PMID- 14530665 TI - Mucosal plication (Gant-Miwa procedure) with anal encircling for rectal prolapse- a review of the Japanese experience. AB - Although mucosal plication for rectal prolapse, known as the Gant-Miwa procedure, is described in some English textbooks, it has been infrequently performed in the West. However, this procedure has been used and developed in conjunction with anal encircling in Japan since the 1960s and is still considered to play a major role in the treatment of rectal prolapse. Certain technical details have been found necessary to ensure the success of the procedure, especially in the technique of anal encircling. For example, the use of Teflon tape and routing relatively deeply and outside the external anal sphincter are necessary. Clinical results show a recurrence rate of 0 to 31 percent with no mortality and almost never any serious complications such as significant bleeding or severe sepsis, which are occasionally encountered in other perineal procedures. Most patients report improved continence after this procedure, and worsening of evacuation is rarely encountered based on our experience. Some physiologic studies have shown improved resting pressure and rectal sensation, which can have a positive influence on the defecatory function. We believe that the Gant-Miwa procedure with anal encircling should be considered as a treatment of choice among perineal procedures for rectal prolapse. PMID- 14530666 TI - Clinical significance of early-onset "sporadic" colorectal cancer with microsatellite instability. PMID- 14530667 TI - A prospective, randomized, multicenter, controlled study of the safety of Seprafilm adhesion barrier in abdominopelvic surgery of the intestine. AB - INTRODUCTION: Seprafilm adhesion barrier (Seprafilm) has been proven to prevent adhesion formation after abdominal and pelvic surgery. This article reports safety results, including the postoperative incidence of abdominal and pelvic abscess and pulmonary embolism, from a large, multicenter trial designed to evaluate the safety and effectiveness of Seprafilm for reduction of adhesion related postoperative bowel obstruction after abdominopelvic surgery. METHODS: A total of 1791 patients participated in this prospective, randomized, multicenter, multinational, single-blind, controlled study in patients undergoing abdominopelvic surgery, the majority of whom had inflammatory bowel disease. Just before closure of the abdomen, patients were randomized to a Seprafilm or no treatment control group. Patients received an average of 4.4 and as many as 10 Seprafilm adhesion barriers applied to organs and tissue surfaces that sustained direct surgical trauma and to suspected adhesiogenic surfaces. Complications that occurred within the first month after surgery were evaluated. RESULTS: During the safety evaluation period, the difference between the Seprafilm and control groups for the incidence of abscess (4 vs. 3 percent, respectively) or pulmonary embolism (<1 percent in both groups) was not statistically significant (P > 0.05). Foreign body reaction was not reported in either group. Fistula (2 vs. <1 percent) and peritonitis (2 vs. <1 percent) occurred more frequently (P 6 cm (10 vs. 1 percent for smaller tumors) and the presence of an invasive carcinoma (20 vs. 3 percent without invasive carcinoma) were significantly associated with an increased probability of recurrence at five years. CONCLUSION: Providing that adequate intraoperative exposure is obtained and advanced malignant tumors receive immediate secondary treatment, transanal resection of clinically benign, large rectal villous adenomas is safe and effective. It is an alternative to rectal resection, which exposes the patient to potentially adverse effects, and also to destructive procedures, which preclude any histologic evaluation of the tumor. PMID- 14530674 TI - What do patients want? Patient preferences and surrogate decision making in the treatment of colorectal cancer. AB - PURPOSE: Clinicians often make decisions for their patients, despite evidence that suggests that correspondence between patient and clinician decision making is poor. The management of colorectal cancer presents difficult decisions because the impact of treatment on quality of life might overshadow its survival efficacy. This study investigated whether patients are able to trade survival for quality of life as a means to express their preference for treatment options and to compare their preferences with those expressed by clinicians. METHODS: Patients undergoing curative surgery for colorectal cancer were interviewed postoperatively to elicit their preferences in four hypothetical treatment scenarios. A questionnaire was mailed to all Australian colorectal surgeons and medical oncologists that asked them to respond as if they themselves were patients. RESULTS: One hundred patients (91 percent), 43 colorectal surgeons (77 percent), and 103 medical oncologists (50 percent) participated. In all four scenarios, patients were able to trade survival for quality of life. Patients' responses varied between scenarios, both in willingness to trade and the average amount traded. There were significant differences between patients and clinicians. Clinicians were more willing than patients to trade survival to avoid a permanent colostomy in favor of chemoradiotherapy. Patients' strongest preference was to avoid chemotherapy, more than to avoid a permanent colostomy. CONCLUSIONS: Patients are able to trade survival as a measure of preference for quality of life and can do so differentially between treatment scenarios. Patients' preferences do not always accord with those of clinicians. Unless patients' preferences are explicitly sought and incorporated into clinical decision making, patients may not receive the treatment that is best for them. PMID- 14530675 TI - Postoperative follow-up of anal condylomata acuminata in HIV-positive patients. AB - PURPOSE: The purpose of this study was to analyze the postoperative follow-up of HIV-positive patients with anal condylomata acuminata, associating recurrence to the AIDS status. METHODS: Ninety-seven male, homosexual patients with anal condylomata were submitted to surgical treatment from August 1992 to December 1997. Specimens were obtained for histologic investigation of Ki-67 cell proliferation marker and for polymerase chain reaction to define the human papillomavirus type. The patients were advised to return weekly during the first month, and monthly up to the sixth month, to evaluate recurrence. Patients with high-grade anal intraepithelial neoplasia remain in follow-up. RESULTS: Histology revealed low-grade anal intraepithelial neoplasia in 81.44 percent of patients and high-grade lesions in 18.56 percent. The results showed that high-grade lesions were not more frequent in late-stage AIDS patients. Ki-67 expression, a cell proliferation marker, was greater in high-grade than in low-grade anal intraepithelial neoplasia, but had no association with AIDS status. Nononcogenic human papillomavirus 6 and 11 were the commonest types. Comparing papillomavirus types and anal intraepithelial grade, we noticed that both oncogenic and nononcogenic types were responsible for high-grade lesions. All patients healed and 51 (52.6 percent) had recurrence up to the sixth month. AIDS status and papillomavirus type were not associated with recurrence. However, high-grade anal intraepithelial neoplasia patients had more recurrence than those with low-grade lesions. Topical treatment failed in 20 patients (41.6 percent), and these were submitted to new surgical treatment. All of them were in the late stage of AIDS. Three who had high-grade lesions in the first operation had low-grade lesions in specimens from the second surgery. The same histologic pattern was observed in 17 patients who had low-grade lesions in warts removed in the first operation. Other patients with high-grade lesions had no recurrence or evolution to invasive carcinoma from five to ten years of follow-up. CONCLUSION: High-grade anal intraepithelial neoplasia and late-stage AIDS are risk factors for recurrence of anal condylomata. PMID- 14530676 TI - Effect of perineal compression on the rectal tone: a study of the mechanism of action. AB - PURPOSE: Digital pressure on the perineum was reported to result in an increase of the rectal tone. This effect has been related to a reflex action named perineorectal reflex but was not verified. The mechanism of action of perineal pressure on the rectal tone was studied. METHODS: Eighteen healthy volunteers (mean age +/- standard deviation, 39.7 +/- 11.8 years; 10 males) were studied. The barostat system used consisted of a polyethylene balloon connected to a strain gauge and a computer-controlled, air-injection system. The balloon was introduced into the rectum, and the rectal tone was assessed by recording the balloon volume variations in response to digital pressure on the perineum. The test was repeated after individual anesthetization of perineum and rectum. It was performed again using normal saline instead of Xylocaine. RESULTS: During perineal pressure, all the volunteers exhibited rectal tone increase with a mean decrease in the balloon volume of 72.3 +/- 14.7 percent. There was no significant difference (P > 0.05) in the rectal tone response between females and males nor between young and elderly patients. The mean latency was 17.3 +/- 1.8 ms. Perineal pressure 20 minutes after individual anesthetization of perineum and rectum effected no significant rectal tone changes. The response returned after the anesthetic effect had waned. The rectal tone response after saline administration was similar to that before administration. CONCLUSION: The study has shown that rectal tone increase during digital perineal pressure represents most probably a reflex action. This was evidenced by absence of rectal tone response on individual anesthetization of the assumed two arms of the reflex arc: perineum and rectum. The perineorectal reflex may be of diagnostic significance in rectal motor disorders and has the potential to be used as an investigative tool, provided further studies are performed to prove these points. PMID- 14530677 TI - Multidimensional analysis of learning curves in laparoscopic sigmoid resection: eight-year results. AB - PURPOSE: The number of operations to reach a plateau phase in colorectal laparoscopic surgery is still under debate. There are few publications reporting on multidimensional assessment of the learning curve, including operation time and complication and conversion rates. The purpose of this study was to define a multidimensional learning curve for sigmoid resection performed by two surgeons with experience in laparoscopic surgery. METHODS: Between 1993 and 2001 from a total of 715 laparoscopic colorectal procedures, two surgeons performed self educated 338 sigmoid resections. Demographic data, indications for surgery, procedure performed, operation time, frequency and kind of complications, conversion rate, and days to discharge were recorded. The moving average method was used to demonstrate the change of the operation time. The cumulative sum technique was used to chart the changes in the conversion and complication rates. RESULTS: Surgeon A performed 199 and Surgeon B 139 sigmoid resections. The operation time decreased from 225 minutes to 169 minutes after approximately 90 operations for Surgeon A and from 270 minutes to 223 minutes after 110 operations for Surgeon B. Based on a decline in intraoperative complications and conversion rate, the steady state was reached after approximately 70 to 80 interventions for both surgeons. CONCLUSIONS: The assessment of a learning curve should not be limited to measurement of a decrease in operation time but should also include the conversion and complication rates. The cumulative sum technique and moving average method as proposed in this study seem appropriate to evaluate the learning curve in this clinical domain. Our findings might be especially useful for those planning training programs in laparoscopic surgery. PMID- 14530679 TI - Randomized, clinical trial of Ligasure vs conventional diathermy in hemorrhoidectomy. AB - PURPOSE: Hemorrhoidectomy is frequently associated with significant postoperative pain, and new techniques to reduce this pain are constantly under evaluation. The present study was conducted to determine the usefulness of the Ligasure system and compare it with conventional diathermy for hemorrhoidectomy. METHODS: Thirty four consecutive patients with Grade 3 or 4 hemorrhoids requiring surgery were recruited and randomized into two groups by preoperative assignment of sealed envelopes. Patients with coexisting perianal disease, previous perianal surgery, or thrombosed hemorrhoids were excluded. All patients were anesthetized and operated on by a single team. In one group, monopolar diathermy in the coagulation mode was used to dissect hemorrhoidal tissue from the internal sphincter. In the second group, tissue was coagulated by Ligasure and then divided with scissors. Operating time was documented by theater staff. Postoperatively, pain scores and Cleveland Clinic incontinence scores were documented. RESULTS: Seventeen patients were randomized into each group. There were no significant differences in age, gender, or clinical symptoms between the groups. The mean operating time in the Ligasure group was 6 (range, 4-10) minutes compared with 11 (range, 7-20) minutes in the other group, and this was statistically significant (P < 0.001). Patients in the Ligasure group reported significantly less pain with first defecation and at postoperative Days 1 and 14 (P < 0.001). The mean hospital stay in both groups was one (range, 1-5) day, and there was no difference in the incontinence scores. CONCLUSIONS: Ligasure diathermy provides a superior alternative to conventional diathermy in hemorrhoidectomy by reducing operating time and postoperative pain. PMID- 14530680 TI - Prospective study comparing multislice CT colonography with colonoscopy in the detection of colorectal cancer and polyps. AB - PURPOSE: Multislice CT colonography is an alternative to colonoscopy. The purpose of this study was to compare multislice CT colonography with colonoscopy in the detection of colorectal polyps and cancers. METHODS: Between June 2000 and December 2001, 45 males and 35 females (median age, 68 (29-83) years) with symptoms of colorectal disease were studied prospectively. All patients underwent multislice CT colonography and colonoscopy, and the findings were compared. RESULTS: Colonoscopy was incomplete in 18 (22 percent) patients because of obstructing lesions or technical difficulty, and multislice CT colonography was unsuccessful in 4 (5 percent) because of fecal residue. Colonoscopy was normal in 26 patients and detected 29 colorectal cancers and 33 polyps in 35 patients, diverticulosis in 16 patients, and colitis in 3 patients. Multislice CT colonography identified 28 of 29 colorectal cancers with one false negative and one false positive (sensitivity, 97 percent; specificity, 98 percent; positive predictive value, 96 percent; negative predictive value, 98 percent). Multislice CT colonography identified all 12 polyps measuring >or=10 mm in diameter (sensitivity, 100 percent), 5 of 6 measuring 6 to 9 mm in diameter (sensitivity, 83 percent), 8 of 15 polyps or=6 mm is similar to colonoscopy. Multislice CT colonography allows clinical staging of colorectal cancers, outlines the whole length of the colon in obstructing carcinoma when colonoscopy fails, and can identify extracolonic causes of abdominal symptoms. PMID- 14530681 TI - Reduced survival of rectal cancer patients with increased tumor epidermal growth factor receptor levels. AB - PURPOSE: The epidermal growth factor receptor and its various ligands (epidermal growth factor, transforming growth factor-alpha, amphiregulin, heparin-binding epidermal growth factor, heregulin, and betacellulin) have been implicated in growth and regeneration of intestinal mucosa and might be related to the development and progression of gastrointestinal tumors. Although some studies have investigated levels of epidermal growth factor receptor by radioligand binding studies, none of them have further analyzed these levels in patients with rectal cancer and investigated their prognostic value. METHODS: We quantitatively determined tumor epidermal growth factor receptor levels in 38 patients with colorectal cancer compared with adjacent normal mucosa by iodine-125-labeled epidermal growth factor binding studies and Scatchard analysis. Patients were followed up for 49.5 +/- 32.2 (range, 2-120) months. RESULTS: Epidermal growth factor receptor capacity was increased in invasive colorectal carcinomas according to T classification (P < 0.001), tumors with lymph node infiltration (P = 0.038), and advanced International Union Against Cancer stage (P < 0.001). Survival of colorectal cancer was reduced in patients with advanced International Union Against Cancer stage (P < 0.001), tumors with positive lymph nodes (P < 0.001), and tumors with elevated epidermal growth factor receptor levels (P = 0.024). In rectal cancer patients, poor prognosis was associated with advanced International Union Against Cancer stage (P = 0.029), tumors with lymph node infiltration (P = 0.040), and increased epidermal growth factor receptor levels (P = 0.002). Multivariate Cox regression analysis indicated that elevated levels of epidermal growth factor receptor were an independent predictor of reduced survival in patients with rectal cancer (P = 0.005). CONCLUSION: The epidermal growth factor receptor/ligand system appears to be involved in tumor development and tumor progression of colorectal carcinomas, with prognostic implication especially in patients with invasive rectal carcinomas. These patients might take advantage of therapies that specifically block epidermal growth factor receptor mediated signal transduction. PMID- 14530682 TI - Budding is useful to select high-risk patients in stage II well-differentiated or moderately differentiated colon adenocarcinoma. AB - PURPOSE: Budding at the invasive front of tumors has recently been thought to suggest greater malignant potential of the colorectal carcinoma. We examined whether budding is associated with poor prognosis after curative resection in Stage II and Stage III colon carcinoma. Furthermore, we examined whether budding is useful to select high-risk patients in Stage II colon carcinoma. METHODS: Surgically resected specimens of 196 Stage II and Stage III colon carcinomas were studied. All the resections were curative (R0), and the median postoperative follow-up was 75.5 months. Using hematoxylin-eosin-stained sections, we determined the presence or absence of budding according to Morodomi's criteria. Routine pathologic findings were also recorded. RESULTS: Budding was detected significantly more frequently in lesions with lymph node metastasis (Stage III) than in lesions without it (Stage II; P < 0.0001). Patients with budding-positive lesions had worse outcome than those with budding-negative lesions: 43 patients (50.6 percent) with budding-positive lesions and 9 (8.1 percent) with budding negative lesions developed recurrence (P < 0.0001). Patients with budding positive lesions had a worse prognosis than patients without it (P < 0.0001). Moreover, no significant difference in survival curves was observed between patients with budding-positive Stage II lesions and those with Stage III lesions (P = 0.930). Multivariate analysis revealed budding as the significant prognostic cofactor of postoperative survival in Stage II and Stage III colon carcinoma (P < 0.0001). CONCLUSION: Budding is useful to select high-risk patients in Stage II colon carcinoma. PMID- 14530683 TI - Prospective comparison of hydrogen peroxide-enhanced three-dimensional endoanal ultrasonography and endoanal magnetic resonance imaging of perianal fistulas. AB - PURPOSE: This study was conducted to determine agreement between hydrogen peroxide-enhanced three-dimensional endoanal ultrasonography and endoanal magnetic resonance imaging in the preoperative assessment of perianal fistulas and to compare these results with the surgical findings. METHODS: Twenty-one patients (aged 26-71 years) with clinical symptoms of a cryptoglandular perianal fistula and a visible external opening underwent preoperative hydrogen peroxide enhanced three-dimensional endoanal ultrasonography, endoanal magnetic resonance imaging, and surgical exploration. The results were assessed separately by experienced observers blinded as to each other's findings. Each fistula was described with notice of the following characteristics: classification of the primary fistula tract according to Parks (intersphincteric, transsphincteric, extrasphincteric, or suprasphincteric), horseshoe, or not classified; presence of secondary tracts (circular or linear); and location of an internal opening. RESULTS: The median time between hydrogen peroxide-enhanced three-dimensional endoanal ultrasonography and endoanal magnetic resonance imaging was 66 (interquartile range, 21-160) days; the median time between the last study (hydrogen peroxide-enhanced three-dimensional endoanal ultrasonography or endoanal magnetic resonance imaging) and surgery was 154 (interquartile range, 95 189) days. Agreement for the classification of the primary fistula tract was 81 percent for hydrogen peroxide-enhanced three-dimensional endoanal ultrasonography and surgery, 90 percent for endoanal magnetic resonance imaging and surgery, and 90 percent for hydrogen peroxide-enhanced three-dimensional endoanal ultrasonography and endoanal magnetic resonance imaging. For secondary tracts, agreement was 67 percent for hydrogen peroxide-enhanced three-dimensional endoanal ultrasonography and surgery, 57 percent for endoanal magnetic resonance imaging and surgery, and 71 percent for hydrogen peroxide-enhanced three dimensional endoanal ultrasonography and endoanal magnetic resonance imaging in case of circular tracts and 76 percent, 81 percent, and 71 percent, respectively, in case of linear tracts. Agreement for the location of an internal opening was 86 percent for hydrogen peroxide-enhanced three-dimensional endoanal ultrasonography and surgery, 86 percent for endoanal magnetic resonance imaging and surgery, and 90 percent for hydrogen peroxide-enhanced three-dimensional endoanal ultrasonography and endoanal magnetic resonance imaging. CONCLUSIONS: For evaluation of perianal fistulas, hydrogen peroxide-enhanced three-dimensional endoanal ultrasonography and endoanal magnetic resonance imaging have good agreement, especially for classification of the primary fistula tract and the location of an internal opening. These results also show good agreement compared with surgical findings. Therefore, hydrogen peroxide-enhanced three-dimensional endoanal ultrasonography and endoanal magnetic resonance imaging can both be used as reliable methods for preoperative evaluation of perianal fistulas. PMID- 14530684 TI - Internal hernia as a complication of colonoscopy: report of a case. AB - After screening colonoscopy, a 65-year-old female developed small-bowel obstruction as demonstrated on abdominal CT scan. The obstruction was caused by a large herniation of the small intestine seemingly caused by the colonoscopic procedure. This is only the second report in the literature of incarcerated intestinal hernia as a complication of colonoscopy. PMID- 14530685 TI - Anal transitional zone cancer after restorative proctocolectomy and ileoanal anastomosis in familial adenomatous polyposis: report of two cases. AB - PURPOSE: Restorative proctocolectomy with ileal pouch-anal anastomosis is accepted as the surgical treatment of choice for many patients with familial adenomatous polyposis. The risk of cancer developing in the ileal pouch after this surgery is unknown. Cancer may arise from the ileal pouch after restorative proctocolectomy, but that arising from the anal transitional zone has not been documented in familial adenomatous polyposis. We report two cases of this cancer from the anal transitional zone in patients with familial adenomatous polyposis, with a review of the literature. METHODS: All patients with familial adenomatous polyposis treated with restorative proctocolectomy and ileal pouch-anal anastomosis in The Cleveland Clinic were included in the study. Patients whose surveillance biopsy of the anal transitional zone revealed invasive adenocarcinoma were studied. RESULTS: Among a total of 146 patients with familial adenomatous polyposis who underwent restorative proctocolectomy and ileal pouch anal anastomosis from 1983 to 2001 in our institution, none developed cancer of the anal transitional zone at up to 18 years of follow-up. However, there were two patients, both of whom underwent surgery elsewhere but who were followed up here, who developed invasive adenocarcinoma of the anal transitional zone. In one of them, cancer was diagnosed three years after a double-stapled ileal pouch-anal anastomosis, whereas in the other, cancer occurred eight years after a straight ileoanal anastomosis with mucosectomy. CONCLUSIONS: Cancer may develop in the anal transitional zone after restorative proctocolectomy with ileal pouch-anal anastomosis for familial adenomatous polyposis. Long-term surveillance of the anal transitional zone needs to be emphasized. PMID- 14530687 TI - Scirrhous colonic metastasis from ductal carcinoma of the breast: report of a case. AB - Metastasis of breast cancer to the colon is rare. We report a case of a 49-year old female who presented with a stenotic tumor of the descending colon five years after treatment of breast cancer with mastectomy and pedicled transverse rectus abdominis musculocutaneous flap. Laparotomy showed a diffusely infiltrated tumor over the descending colon. Anterior resection with loop ileostomy was performed, and the pathology showed that the colonic wall and the mesentery were diffusely infiltrated with poorly differentiated adenocarcinoma, which stained strongly for cytokeratin 7. The histologic diagnosis is consistent with colonic metastasis from ductal carcinoma of breast origin. In a patient with a history of breast cancer, colonic metastasis from the breast primary cancer should be considered, especially if the colonic lesion is scirrhous in nature. The incision for laparotomy and the probable stoma site should be planned carefully in females after breast reconstructive surgery. PMID- 14530688 TI - Save the skin and improve the scar: a simple technique to minimize the scar from a temporary stoma. AB - Reversal of a loop ileostomy is not infrequently associated with wound infection and delayed wound healing. This in turn may lead to a disappointing scar. A simple modification to the technique of stoma construction simplifies wound closure, reduces the rate of wound morbidity, and improves the cosmetic result. PMID- 14530689 TI - Usefulness of flexible colonoscopic microwave coagulation therapy for a colorectal anastomotic stricture. AB - PURPOSE: Several methods of treatment for benign anastomotic strictures after anterior resection have been described. We describe a simple, safe, effective, and inexpensive method for treating benign colorectal anastomotic stricture by means of microwave coagulation under flexible colonoscopic visualization. METHODS: Eighteen patients with rectal or rectosigmoidal cancer underwent low anterior resection or anterior resection without colonic pouch and colorectal anastomosis by a double-stapling technique with PCEEA. Two of 18 patients (11.1 percent) developed an anastomotic stenosis. A microwave electrode was passed through the biopsy channel of the flexible colonoscope. Under flexible colonoscopic visualization, microwave irradiation was performed at four points (3, 6, 9, and 12 o'clock) in the stricture site because of granulation scar to obtain an adequately coagulated area. No bougies were performed thereafter. RESULTS: The anastomotic strictures could be dilated adequately, the patients could defecate satisfactorily, and their abdominal distentions and bowel symptoms were resolved. No complications occurred. No occurrence of restricture has been observed. CONCLUSION: Flexible endoscopic, microwave coagulation therapy is a useful, simple, effective, and safe method for the treatment of benign colorectal anastomotic strictures. PMID- 14530690 TI - Improved quality of anorectal endoluminal ultrasonography using emulsion of dimethicone. AB - PURPOSE: This study was designed to show the benefits of filling echography probes with water mixed with dimethicone to preclude artifacts caused by bubbles. METHODS: One hundred consecutive healthy volunteers (42 males; mean age, 46 years; range, 17-72 years) were blindly randomized to undergo rectal ultrasonography using conventional degassed water or degassed aqueous solution made of 40 ml of water and 10 ml of dimethicone emulsion (1 ml/100 mg). All examinations performed by the same surgeon-unaware of nature of filler liquid-for ten minutes, and number of rectal ultrasonographies with artifacts attributable to air bubbles was recorded. RESULTS: Endoluminal ultrasonography performed with conventional degassed water presented artifacts attributed to presence of air bubbles in 30 of 50 examinations (60 percent). When degassed aqueous solution was used, only 5 of 50 examinations showed artifacts caused by presence of air bubbles (10 percent). This difference was statistically significant (P < 0.0005). The use of conventional fluid was associated with artifacts secondary to the presence of air bubbles, adjusted odds ratio 13.5 (95 percent confidence interval, 4.56-39.88). During this experience, the solution did not damage ultrasound scanner. CONCLUSIONS: We found that use of a dimethicone-degassed water mixture is simple, not expensive, safe, effective, and may reduce frequency of sonographic distortion in presence of air bubbles. PMID- 14530691 TI - Influence of school bag carrying on gait kinetics. AB - The purpose of this study was to determine the effect of different methods of backpack carrying on gait kinetics in children, using a new treadmill that allowed three-dimensional measurement of right and left leg ground reaction forces (GRFs). Forty-one healthy children, with a mean age of 12 years, participated in this study. The mean height was 152 cm and the mean weight 40 kg. The three trials consisted of walking on the treadmill at the speed of 3.5 km/h, first without a backpack and then carrying a 10 kg school bag on the right shoulder or on both shoulders. For each carrying condition GRFs were recorded, averaged, and analyzed for 30 steps. Stride, stance, double stance, thirteen specific GRF parameters and the symmetry index were measured. The right leg produced higher propulsive fore-aft forces than the left one, whatever the walking conditions. For the two maximum peaks and the average vertical force during stance, a statistical difference was found between walking without a backpack and carrying a backpack on one or two shoulders (one or two shoulder carrying > no backpack) but never between one-shoulder and two-shoulder carrying. The children increased their stance and double stance when walking with a backpack compared with walking without a pack. The symmetry index increased with one-strap carrying (compared with no backpack and two-strap carrying) for the maximum force during the breaking phase (Fy1) when it decreased for the maximum propulsive horizontal force before taking-off (Fy2). Children should be advised to carry their backpack on two shoulders rather than use a one-strap backpack. PMID- 14530692 TI - A sonographic study of perinatal hip development: from 34 weeks of gestation to 6 weeks of age. AB - A sonographic study of perinatal hip development was performed by consecutive measurement of Graf's alpha and beta angles in fetal and newborn hips. The study group consisted of infants with sonographically normal hip findings at birth. Forty fetuses were examined by fetal hip sonography at 34, 36 and 38 weeks of gestation. Postnatally, hip sonography was performed in the first and sixth week of age. Maturation curves of the bony (alpha-angle) and cartilaginous (beta angle) acetabular roof from 34 weeks of gestation to 6 weeks of age were established. Prenatally, the mean alpha-angles were above the level that corresponds to a mature hip joint. A significantly higher value of the mean alpha angles was found after birth. The mean beta-angles of the fetuses did not differ from those of the newborns. Our results revealed that the fetal hip joint is sonographically mature at 34 weeks of gestation. Further progression of hip development occurs around term. PMID- 14530693 TI - Body mass index and slipped capital femoral epiphysis. AB - Body mass index (BMI) as a predictor of slipped capital femoral epiphysis (SCFE) was studied. A total of 26 adolescent patients had complete annual height and weight measurements taken from birth to onset of slippage. These values were compared with those of the normal adolescent population. Patients with SCFE showed statistically higher BMI during growth than normal developing children. BMI gives more accurate data on body build than height and weight alone and may be a useful tool for evaluating risk factors in SCFE. PMID- 14530694 TI - Pinning in slipped capital femoral epiphysis: long-term follow-up study. AB - This long-term follow-up study was aimed at evaluating patients with hip epiphysiolysis treated by pinning and previously controlled by short-term follow up, in order to evaluate both clinical and radiographic evolution. The case series included 36 patients (17 female and 19 male) for overall 48 hips (21 right, 27 left). The epiphysiolysis was bilateral in 12 patients. Median follow up was 17.9 years (range 8.8-29.2). Clinical results were evaluated according to Harris, radiographic results according to Boyer. The patients were divided into three groups according to slipping degree calculated following Southwick (0-30 degrees, 30-60 degrees, >60 degrees ). Slipping degree was found to be directly correlated with worsening of results. Reduction manoeuvres on chronic epiphysiolysis proved to lead to even poorer results. Our review, even though carried out on a limited number of cases, showed that (1) hips with arthrosis at first control resulted in worsening in the majority of cases; (2) slipping degree was proportional to the result obtained; (3) reduction manoeuvres performed on chronic epiphysiolysis had a negative influence on results; (4) the presence of chondrolysis or epiphyseal necrosis always led to early hip arthrosis; (5) in the absence of major complications, evolution towards arthrosis can be slow and adulthood or even old age can be reached without resorting to total hip replacement. PMID- 14530695 TI - Capital femoral epiphysis and growth plate of the asymptomatic hip joint in unilateral Perthes disease. AB - Radiograms of 173 children with unilateral Perthes disease were reviewed with particular reference to the presence of flattening and irregularity of the 'unaffected' epiphyses. The slope and shape of the growth plate were also evaluated. About 35% of children with Perthes disease showed changes of the unaffected proximal femur in the first radiograms of the hip joints. These consisted of small epiphysis, flattening of the epiphysis, contour irregularities (20%), and changes in the growth plate (15%). The radiographic abnormalities were not accompanied by any clinical symptoms of the asymptomatic hip joint and they receded with age during treatment. PMID- 14530696 TI - Triple pelvic osteotomy in Legg-Calve-Perthes disease using a single anterolateral incision. AB - Pelvic osteotomy has a small role to play in the management of Legg-Calve-Perthes disease. Residual acetabular dysplasia or incongruity can be corrected by rotational acetabuloplasty. In skeletally immature patients this can be performed by complete or incomplete innominate osteotomies. We present the case of a 12 year-old child with residual acetabular dysplasia, resulting from Legg-Calve Perthes disease, which was treated by a triple pelvic osteotomy performed by a small single anterolateral incision. PMID- 14530697 TI - Arthroscopic meniscectomy for discoid lateral meniscus in children and adolescents: 4.5 year follow-up. AB - We retrospectively reviewed the 4.5 year (range, 21-88 months) follow-up results of arthroscopic partial meniscectomies performed in 11 knees between 1994 and 2000 to treat a symptomatic discoid lateral meniscus. The average age at surgery was 11.5 years (range, 5-17 years). All except one of the discoid menisci were of a complete type, and all except three were torn menisci. At the latest follow-up examination, the result was excellent for nine knees, and good for two; no degenerative changes were evident on the roentgenograms. Arthroscopic partial meniscectomy should be the treatment of choice for the complete type symptomatic lateral discoid meniscus, even if it is intact. Preoperative lack of the knee extension requires a gentle rehabilitation program postoperatively. PMID- 14530698 TI - Radial dysplasia with localized cutis aplasia congenita. AB - We describe four cases of distal radial epiphyseal dysplasia associated with a localized area of cutis aplasia congenita over the dorsum of the distal forearm. The cutis aplasia was diagnosed at birth in all cases, but the radial dysplasia was not recognized until presentation to our orthopaedic department between the ages of 5 and 10 years. This association, the long-term implications and the requirement for follow-up until skeletal maturity have not previously been described. We emphasize the importance of continued monitoring of these patients as the effects of radial dysplasia did not become obvious for several years, and the potential benefit from achieving early skin cover with grafting rather than allowing healing by secondary intention is discussed. It is important to increase awareness of this condition so that early orthopaedic and plastic surgical opinions can be sought, in order to reduce the disabling effect on the underlying radius. In this way it may be possible to elucidate the cause and manage the condition satisfactorily at an earlier stage. PMID- 14530699 TI - Avascular necrosis of the intermediate cuneiform bone. AB - We present a case of avascular necrosis of the intermediate cuneiform bone in a skeletal immature patient with a spontaneous recovery following conservative treatment. We discuss the aetiology and options of treatment. PMID- 14530700 TI - The triplane variant of the tibial apophyseal fracture: a case report and a review of the literature. AB - A rare case of tibial apophyseal fracture with extension in the antero-lateral physis and epiphysis in a 17-year-old boy is described. This type of fracture, which can be associated with intra-articular lesions and lesions of the extensor mechanism, resembles the better known distal tibial triplane fracture. Arthroscopic controlled anatomic reduction with restoration of the articular surface was achieved. PMID- 14530701 TI - BSN-to-PhD programs: a bad idea. PMID- 14530702 TI - Where's the evidence? PMID- 14530703 TI - Government and health care. PMID- 14530704 TI - Recognizing APRNs. PMID- 14530706 TI - Emergency: airway management. PMID- 14530707 TI - Traditional healing and HIV-AIDS in KwaZulu-Natal, South Africa. PMID- 14530708 TI - Erectile dysfunction. PMID- 14530709 TI - Epidural analgesia for postoperative pain. PMID- 14530711 TI - Work and no pay? PMID- 14530712 TI - Looks can be deceiving. PMID- 14530713 TI - Telephonic nursing in traumatic brain injury. PMID- 14530714 TI - "Horizontal hostility". PMID- 14530715 TI - Catering to the homeless in Atlanta. Interview by Joy Jacobson and Ali Fenwick. PMID- 14530716 TI - Preparing for a possible SARS resurgence. PMID- 14530717 TI - The Leapfrog volume criteria may fall short in identifying high-quality surgical centers. AB - OBJECTIVE: The original Leapfrog Initiative recommends selective referral based on procedural volume thresholds (500 coronary artery bypass graft [CABG] surgeries, 30 abdominal aortic aneurysm [AAA] repairs, 100 carotid endarterectomies [CEA], and 7 esophagectomies annually). We tested the volume mortality relationship for these procedures in the University HealthSystem Consortium (UHC) Clinical DatabaseSM, a database of all payor discharge abstracts from UHC academic medical center members and affiliates. We determined whether the Leapfrog thresholds represent the optimal cutoffs to discriminate between high- and low-mortality hospitals. METHODS: Logistic regression was used to test whether volume was a significant predictor of mortality. Volume was analyzed in 3 different ways: as a continuous variable, a dichotomous variable (above and below the Leapfrog threshold), and a categorical variable. We examined all possible thresholds for volume and observed the optimal thresholds at which the odds ratio is the highest, representing the greatest difference in odds of death between the 2 groups of hospitals. RESULTS: In multivariate analysis, a relationship between volume and mortality exists for AAA in all 3 models. For CABG, there is a strong relationship when volume is tested as a dichotomous or categorical variable. For CEA and esophagectomy, we were unable to identify a consistent relationship between volume and outcome. We identified empirical thresholds of 250 CABG, 15 AAA, and 22 esophagectomies, but were unable to find a meaningful threshold for CEA. CONCLUSIONS: In this group of academic medical centers and their affiliated hospitals, we demonstrated a significant relationship between volume and mortality for CABG and AAA but not for CEA and esophagectomy, based on the Leapfrog thresholds. We described a new methodology to identify optimal data based volume thresholds that may serve as a more rational basis for selective referral. PMID- 14530718 TI - Clinical and pathologic response of Barrett's esophagus to laparoscopic antireflux surgery. AB - BACKGROUND DATA: Patients with Barrett's esophagus (BE) are frequently offered laparoscopic antireflux surgery (LARS) to treat symptoms. The effectiveness of this operation with regards to symptoms and to the evolution of the columnar lined epithelium remains controversial. METHODS: We analyzed the course of 106 consecutive patients with BE who underwent LARS between 1994 and 2000, representing 14% of all LARS (754) performed in our institution during that period. All 106 patients agreed to clinical follow-up in 2002 at 40 months (median; range, 12-95 months). Fifty-three patients (50%) agreed to functional evaluation (manometry and 24-hour pH monitoring); 90 patients (85%) to thorough endoscopy, with appropriate biopsies and histologic evaluation to determine the status of BE. RESULTS: Heartburn improved in 94 (96%) of 98 and resolved in 69 patients (70%) after LARS. Regurgitation improved in 58 (84%) of 69 and dysphagia improved in 27 (82%) of 33. Distal esophageal acid exposure improved in 48 (91%) of 53 patients tested and returned to normal in 39 patients (74%). One patient underwent reoperation 2 days after fundoplication (gastric perforation). Preoperatively, biopsy revealed BE without dysplasia in 91 patients, BE indefinite for dysplasia in 12 patients, and low-grade dysplasia in 3 patients. Fifty-four of the 90 patients with endoscopic follow-up had short-segment BE (<3cm), and 36 had long-segment BE (>3cm) preoperatively. Postoperatively, endoscopy and pathology revealed complete regression of intestinal metaplasia (absence of any sign suggestive of BE) in 30 (55%) of 54 patients with short segment BE but in 0 of 36 of those with long-segment BE. Among patients with complete regression, 89% of those tested with pH monitoring had normal esophageal acid exposure. This was observed in 69% of those who failed to have complete regression. One patient developed adenocarcinoma within 10 months of LARS. CONCLUSIONS: In patients with BE, LARS provides excellent control of symptoms and esophageal acid exposure. Moreover, intestinal metaplasia regressed in the majority of patients who had short-segment BE and normal pH monitoring following LARS, a fact that was, heretofore, not appreciated. LARS should be recommended to patients with BE to quell symptoms and to prevent the development of cancer. PMID- 14530719 TI - Effect of laparoscopic Roux-en Y gastric bypass on type 2 diabetes mellitus. AB - OBJECTIVE: To evaluate pre- and postoperative clinical parameters associated with improvement of diabetes up to 4 years after laparoscopic Roux-en-Y gastric bypass (LRYGBP) in patients with type 2 diabetes mellitus (T2DM). SUMMARY BACKGROUND DATA: The surgical treatment of morbid obesity leads to dramatic improvement in the comorbidity status of most patients with T2DM. However, little is known concerning what preoperative clinical factors are associated with postoperative long-term improvement in diabetes in the morbidly obese patient with diabetes. METHODS We evaluated pre- and postoperative data, including demographics, duration of diabetes, metabolic parameters, and clinical outcomes, in all patients with impaired fasting glucose (IFG) and type T2DM undergoing LRYGBP from July 1997 to May 2002. RESULTS: During this 5-year period, 1160 patients underwent LRYGBP and 240 (21%) had IFG or T2DM. Follow up was possible in 191 of 240 patients (80%). There were 144 females (75%) with a mean preoperative age of 48 years (range, 26-67 years). After surgery, weight and body mass index decreased from 308 lbs and 50.1 kg/m2 to 211 lbs and 34 kg/m2 for a mean weight loss of 97 lbs and mean excess weight loss of 60%. Fasting plasma glucose and glycosylated hemoglobin concentrations returned to normal levels (83%) or markedly improved (17%) in all patients. A significant reduction in use of oral antidiabetic agents (80%) and insulin (79%) followed surgical treatment. Patients with the shortest duration (<5 years), the mildest form of T2DM (diet controlled), and the greatest weight loss after surgery were most likely to achieve complete resolution of T2DM. CONCLUSION: LRYGBP resulted in significant weight loss (60% percent of excess body weight loss) and resolution (83%) of T2DM. Patients with the shortest duration and mildest form of T2DM had a higher rate of T2DM resolution after surgery, suggesting that early surgical intervention is warranted to increase the likelihood of rendering patients euglycemic. PMID- 14530720 TI - Minimally invasive esophagectomy: outcomes in 222 patients. AB - OBJECTIVE: To assess our outcomes after minimally invasive esophagectomy (MIE). SUMMARY BACKGROUND DATA: Esophagectomy has traditionally been performed by open methods. Results from most series include mortality rates in excess of 5% and hospital stays frequently greater than 10 days. MIE has the potential to improve these results, but only a few small series have been reported. This report summarizes our experience of 222 cases. METHODS: From 1996 to 2002, MIE was performed in 222 patients. Indications for operation included high-grade dysplasia (n = 47) and cancer (n = 175). Neoadjuvant chemotherapy was used in 78 (35.1%) and radiation in 36 (16.2%). Initially, a laparoscopic transhiatal approach was used (n = 8), but subsequently our approach evolved to include thoracoscopic mobilization (n = 214). RESULTS: There were 186 men and 36 women. Median age was 66.5 years (range, 39-89). Nonemergent conversion to open procedure was required in 16 patients (7.2%). MIE was successfully completed in 206 (92.8%) patients. The median intensive care unit stay was 1 day (range, 1 30); hospital stay was 7 days (range, 3-75). Operative mortality was 1.4% (n = 3). Anastomotic leak rate was 11.7% (n = 26). At a mean follow-up of 19 months (range, 1-68), quality of life scores were similar to preoperative values and population norms. Stage specific survival was similar to open series. CONCLUSIONS: MIE offers results as good as or better than open operation in our center with extensive minimally invasive and open experience. In this single institution experience, we observed a lower mortality rate (1.4%) and shorter hospital stay (7 days) than most open series. Given these results, we are now developing an intergroup trial (ECOG 2202) to assess MIE in a multicenter setting. PMID- 14530721 TI - One hundred in situ split-liver transplantations: a single-center experience. AB - OBJECTIVE: To identify predictors of graft and recipient survival from a single institution series of in situ split-liver transplantations and compare outcomes to living donor and whole organs for adults and children. SUMMARY BACKGROUND DATA: Split-liver transplantation is a surgical technique that creates 2 allografts from a single cadaver donor. We have applied split-liver transplantation to all indications and categories of medical urgency for initial as well as retransplantation to expand the current donor pool and decrease reliance upon living donation. METHODS: A retrospective analysis was conducted of 100 consecutive in situ split-liver transplantations yielding a left lateral segment and right trisegment graft that were performed at the University of California Los Angeles between 9/91 and 02/03. These 100 transplantations generated 190 allografts for transplantation into 105 children and 60 adults, with the sharing of 25 allografts among transplant centers across the United States. Outcomes and incidence of complications were compared with living donor and whole organ recipients receiving liver transplantation during the same time period with independent predictors of split-liver graft and recipient survival identified by multivariate analysis. RESULTS: The incidence of biliary and vascular complications observed in recipients of left lateral segment grafts created by split-liver transplantation was not statistically different from recipients of left lateral segment grafts created from living donation or children receiving whole-organ grafts from pediatric donors. Kaplan-Meier survival estimations of left lateral segment graft and recipient survival also demonstrated no statistical difference among split-liver, living donor, and whole organ recipients. Right trisegment grafts from split-liver transplantation demonstrated a 10% incidence of biliary and 7% incidence of vascular complications. Long-term graft function was excellent with patient and graft survival equal to 1086 recipients of cadaver whole-organ grafts from donors ages 10-40 years who underwent transplant operations during the same time period. Predictors of split-liver transplantation graft and recipient survival included United Network for Organ Sharing status at transplantation, indication, occurrence of a complication, donor creatinine, and donor length of hospitalization. CONCLUSIONS: Split-liver transplantation is an effective mechanism for immediate expansion of the cadaver donor pool that can reduce dependence upon living donation in adults and children. PMID- 14530722 TI - Liver resection as a bridge to transplantation for hepatocellular carcinoma on cirrhosis: a reasonable strategy? AB - OBJECTIVE: To assess the viability of a strategy of primary resection with secondary liver transplantation (LT) for hepatocellular carcinoma (HCC) on cirrhosis. SUMMARY BACKGROUND DATA: LT is the optimal treatment of HCC with cirrhosis. Owing to organ shortage, liver resection is considered as a reasonable first-line treatment of patients with small HCC and good liver function, with secondary LT as a perspective in case of recurrence. The viability of such strategy, positively explored in theoretical models, is not documented in clinical practice. METHODS: Among 358 consecutive patients with HCC on cirrhosis treated by liver resection (n = 163; 98 of whom were transplantable) or transplantation (n = 195), the feasibility and outcome of secondary transplantation was evaluated in a 2-step fashion. First, secondary LT for tumor recurrence after resection (n = 17) was compared with primary LT (n = 195), to assess the risk and the outcome of secondary LT in patients who effectively succeeded to be treated by this approach. Second, primary resection in transplantable patients (n = 98) was compared with that of primary LT (n = 195) on an intention-to-treat basis, to assess the outcome of each treatment strategy and to determine the proportion of resected patients likely to be switched for secondary LT. Transplantability of resected patients was retrospectively determined according to selection criteria of LT for HCC. RESULTS: Operative mortality (< or =2 months) of secondary LT was significantly higher than that of primary LT (28.6% versus 2.1%; P = 0.0008) as was intraoperative bleeding (mean transfused blood units, 20.7 versus 10.5; P = 0.0001). Tumor recurrence occurred more frequently after secondary than after primary LT (54% versus 18%; P = 0.001). Posttransplant 5-year overall survival was 41% versus 61% (P = 0.03), and disease-free survival was 29% versus 58% (P = 0.003) for secondary and primary LT, respectively. Of 98 patients treated by resection while initially eligible for transplantation, only 20 (20%) were secondarily transplanted, 17 of whom (17%) for tumor recurrence and 3 (3%) for hepatic decompensation. Transplantability of tumoral recurrence was 25% (17 of 69 recurrences). Compared with primarily transplanted patients, transplantable resected patients had a decreased 5-year overall survival (50% versus 61%; P = 0.05) and disease-free survival (18% versus 58%; P < 0.0001), despite the use of secondary LT. On a multivariate analysis including 271 patients eligible for transplantation and treated by either liver resection or primary LT, liver resection alone (P < 0.0001; risk ratio [RR] = 3.27) or liver resection with secondary LT (P < 0.05; RR= 1.87) emerged as negative independent factors of disease-free survival as compared with primary LT. A number of nodules > 3 (P = 0.002; RR= 2.02) and a maximum tumor size exceeding 30 mm (P < 0.0001; RR=1.93) were also predictive of lower disease-free survival. CONCLUSIONS: LT after liver resection is associated with a higher operative mortality, an increased risk of recurrence, and a poorer outcome than primary LT. In addition, liver resection as a bridge to LT impairs the patient transplantability and the chance of long-term survival of cirrhotic patients with HCC. Primary LT should therefore remain the ideal choice of treatment of a cirrhotic patient with HCC, even when the tumor is resectable. PMID- 14530723 TI - Kidney transplantation under a tolerogenic regimen of recipient pretreatment and low-dose postoperative immunosuppression with subsequent weaning. AB - OBJECTIVE: The purpose of this work was to perform kidney transplantation under a regimen of immunosuppression that facilitates rather than interferes with the recently defined mechanisms of alloengraftment and acquired tolerance. SUMMARY BACKGROUND DATA: In almost all centers, multiple immunosuppressive agents are given in large doses after kidney transplantation in an attempt to reduce the incidence of acute rejection to near zero. With the elucidation of the mechanisms of alloengraftment and acquired tolerance, it was realized that such heavy prophylactic immunosuppression could systematically subvert the clonal exhaustion deletion that is the seminal mechanism of tolerance. In addition, it has been established that the rejection response can be made more readily treatable by pretransplant immunosuppression. Consequently, we conducted kidney transplantation in compliance with 2 therapeutic principles: recipient pretreatment and the least possible use of posttransplant immunosuppression. METHODS: One-hundred fifty unselected renal transplant recipients with a mean age of 51 +/- 15 years and multiple risk factors had pretreatment with approximately 5 mg/kg of rabbit antithymocyte globulin (Thymoglobulin) in the hours before transplantation, under covering bolus doses of prednisone to prevent cytokine reactions. Minimal posttransplant immunosuppression was with tacrolimus monotherapy to which steroids or other agents were added only for the treatment of rejection. At or after 4 months after transplant, spaced-dose weaning from tacrolimus monotherapy was begun in patients who had exhibited a satisfactory course. RESULTS: One-year actuarial patient and graft survival was 97% and 92%, respectively. Although the incidence of early acute rejection was 37%, only 7% required prolonged treatment with any agent other than tacrolimus. After a follow up of 6 to 21 months, the mean serum creatinine in patients with functioning grafts is 1.8 +/- 1.0 mg/dL. Seventy-three percent of the patients met the criteria for spaced weaning. Although rejection episodes occasionally required restoration of daily treatment, 94 (63%) of the 150 patients currently receive tacrolimus in spaced doses ranging from every other day to once a week. CONCLUSIONS: With this approach to immunosuppression, it has been possible to avoid early posttransplant overimmunosuppression and thereby to promote the evolution of a degree of partial tolerance sufficient to undertake substantial dose reduction. The strategy, which is applicable for all organ grafts, constitutes a paradigm shift in transplant management at our center. PMID- 14530724 TI - Thin melanomas: predictive lethal characteristics from a 30-year clinical experience. AB - OBJECTIVE: To guide treatment and clinical follow-up by defining the natural history of thin melanomas and identifying negative prognostic characteristics that may delineate high-risk patients. SUMMARY BACKGROUND DATA: In following > 10,000 patients with cutaneous melanoma over the past 30 years, our institution has observed nodal or metastatic disease in approximately 15% of patients with a thin (<1 mm) primary lesion. METHODS: A database query of patients with cutaneous melanoma returned 1158 patients with primary lesion < or = 1 mm thick and who received their initial treatment at a single institution. Median follow-up was 11 years (range, 1 to 34 years). Patient and melanoma characteristics as well as outcomes were recorded and statistically analyzed. RESULTS: 6.6% of patients had nodal or distant disease at presentation. Over time, an additional 9.4% developed metastases, including nodal and distal recurrences. Overall incidence of advanced disease was 15.3%. Univariate analysis identified male gender (P = 0.01), advanced age (>45 years; P = 0.05), and Breslow thickness (>0.75 mm; P = 0.008) as significant negative prognostic characteristics. Of patients with these 3 high risk characteristics, 19.7% developed advanced disease (likelihood ratio 6.3; P = 0.007 versus nonhigh-risk patients). This group had more than twice the incidence of nodal recurrences. Patients with recurrence had significantly decreased 10 year survival (82% versus 45%; P < 0.0001). Surprisingly, neither ulceration nor Clark level predicted advanced disease. CONCLUSIONS: Thin melanomas are potentially lethal lesions. Long-term follow-up identified a high-risk population of older males with tumors between 0.75 mm and 1.0 mm whose risk of recurrent disease approaches 20%. Traditionally accepted negative prognostic factors such as ulceration and discordant Clark levels are not predictive for metastasis in this population. Given the poor prognosis associated with recurrent disease, we recommend close clinical evaluation and follow-up to maximize accurate staging and therapeutic options. PMID- 14530725 TI - Lymphatic mapping and sentinel lymphadenectomy for early-stage melanoma: therapeutic utility and implications of nodal microanatomy and molecular staging for improving the accuracy of detection of nodal micrometastases. AB - OBJECTIVE: Lymphatic mapping and sentinel lymphadenectomy (LM/SL) have been applied to virtually all solid neoplasms since our original description of LM/SL for melanoma. Our objectives were to determine the diagnostic and therapeutic utility of LM/SL, investigate carbon dye for mapping the microanatomy of lymphatic flow within the sentinel node (SN), and determine the prognostic accuracy of molecular assessment of the SN. METHODS: Since 1985, 1599 patients with AJCC Stage I/II melanoma have been treated by LM/SL at our institution and 4590 have been treated by wide excision (WE) without nodal staging. We examined the incidence of clinical nodal recurrence after WE alone, the incidence of subclinical nodal metastases found by LM/SL, and the incidence of nodal recurrence in basins with histopathology-negative SNs. RESULTS: In 1514 LM/SL patients with a primary of known Breslow thickness, the incidence of metastasis in nodes claimed to be sentinel was 7.3%, 19.7%, 33.2%, and 39.7% for primary lesions 4.0 mm, respectively. In 3652 WE-only patients, the corresponding rates of nodal recurrence were 12.0%, 32.0%, 34.4%, and 30.1%. Thus, LM/SL detected only 60% of expected nodal metastases from primary melanomas <2.01 mm. Forty of 1599 (3.1%) patients developed recurrence in basins with immunohistochemistry (IH)-negative SNs. To determine whether nonrandom intranodal distribution of tumor cells could explain missed SN metastases, we coinjected carbon particles and blue dye during LM/SL in 166 patients: 25 (16%) patients had nodal metastases, all of which were found only in nodal subsectors containing carbon particles. When paraffin-embedded SNs from a subset of 162 IH-negative patients were re-examined by quantitative multimarker reverse-transcriptase polymerase chain reaction (qRT) assay, 49 (30%) gave positive signals. These patients had a significantly higher risk of disease recurrence and death than did patients whose IH and qRT results were negative (p < 0.0001). Comparison of 287 prognostically matched pairs of patients who underwent immediate (after LM/SL) versus delayed (after observation) dissection of nodal metastases revealed 5-, 10-, and 15-year survival rates of 73%, 69%, and 69% versus 51%, 37%, and 32%, respectively (P < or = 0.001). CONCLUSIONS: SN assessment based on intranodal compartmentalization of lymphatic flow (carbon dye mapping) should increase the accuracy of IH and, in combination with multimarker qRT assessment, will allow confident identification of most patients for whom surgery alone is curative. Our data suggest a significant therapeutic benefit for immediate dissection based on identification of a tumor-involved SN. PMID- 14530726 TI - Natural history of > or =60% asymptomatic carotid stenosis in patients with contralateral carotid occlusion. AB - OBJECTIVE: Although the Asymptomatic Carotid Atherosclerosis Study (ACAS) reported that carotid endarterectomy (CEA) is beneficial for patients with asymptomatic > or =60% carotid stenosis (ACS), several other studies have reported mixed results. Our prospective study analyzed the natural history of > or =60% ACS in patients with contralateral carotid occlusion (CCO). PATIENT POPULATION AND METHODS: During a 10-year period, patients with 60-<70% ACS with CCO were entered into a protocol of clinical examination and duplex surveillance every 6 months. All patients underwent maximum medical therapy. Late CEAs were considered if lesions became symptomatic or progressed to > or =70% stenosis. A Kaplan-Meier lifetable analysis was performed to estimate the freedom from both ipsilateral strokes and all strokes. RESULTS: Eighty-two patients were enrolled with a mean follow-up of 59.5 months (range, 7-141 months). Late strokes were noted in 27 of 82 patients (33%); 19 (23%) were ipsilateral and 8 (10%) were contralateral (side of CCO). Late transient ischemic attacks (TIAs) were noted in 22 of 82 (27%, 7 ipsilateral and 15 contralateral). The combined neurologic event (TIA/stroke) rate was 60% (49 of 82, 32% ipsilateral and 28% contralateral). Kaplan-Meier lifetable analysis showed that the rates of freedom from ipsilateral strokes, all strokes, and progression to > or =70% stenosis at 1, 2, 3, 4, and 5 years were 94%, 90%, 85%, 80%, 73%; 94%, 89%, 84%, 77%, 67%; and 99%, 96%, 92%, 86%, and 82%, respectively. The ipsilateral stroke-free survival rates at l, 2, 3, 4, and 5 years were 94%, 88%, 78%, 70%, and 63%. Twenty-one late CEAs were performed with no perioperative stroke/deaths (5 for ipsilateral TIAs, 9 for ipsilateral strokes, and 7 for > or =70% ACS). Overall, 20 (24%, 11 with symptoms and 9 asymptomatic) progressed to > or =70% stenosis. CONCLUSIONS: Patients with 60-<70% ACS and CCO with maximal medical therapy carry a higher incidence of ipsilateral strokes and all strokes than what was reported by the ACAS study; therefore, prophylactic CEA may be justified in these patients. PMID- 14530727 TI - Ewing sarcoma/primitive neuroectodermal tumor of the chest wall: impact of initial versus delayed resection on tumor margins, survival, and use of radiation therapy. AB - OBJECTIVE: To establish outcome and optimal timing of local control for patients with nonmetastatic Ewing sarcoma/primitive neuroectodermal tumor (ES/PNET) of the chest wall. METHODS: Patients < or =30 years of age with ES/PNET of the chest wall were entered in 2 consecutive protocols. Therapy included multiagent chemotherapy; local control was achieved by resection, radiotherapy, or both. We compared completeness of resection and disease-free survival in patients undergoing initial surgical resection versus those treated with neoadjuvant chemotherapy followed by resection, radiotherapy, or both. Patients with a positive surgical margin received radiotherapy. RESULTS: Ninety-eight (11.3%) of 869 patients had primary tumors of the chest wall. Median follow-up was 3.47 years and 5-year event-free survival was 56% for the chest wall lesions. Ten of 20 (50%) initial resections resulted in negative margins compared with 41 of 53 (77%) negative margins with delayed resections after chemotherapy (P = 0.043). Event-free survival did not differ by timing of surgery (P = 0.69) or type of local control (P = 0.17). Initial chemotherapy decreased the percentage of patients needing radiation therapy. Seventeen of 24 patients (70.8%) with initial surgery received radiotherapy compared with 34 of 71 patients (47.9%) who started with chemotherapy (P = 0.061). If a delayed operation was performed, excluding those patients who received only radiotherapy for local control, only 25 of 62 patients needed radiotherapy (40.3%; P = 0.016). CONCLUSION: The likelihood of complete tumor resection with a negative microscopic margin and consequent avoidance of external beam radiation and its potential complications is increased with neoadjuvant chemotherapy and delayed resection of chest wall ES/PNET. PMID- 14530728 TI - One-stage transanal Soave pullthrough for Hirschsprung disease: a multicenter experience with 141 children. AB - BACKGROUND: The surgical management of Hirschsprung's disease (HD) has evolved from the original 3-stage approach to the recent introduction of minimal-access single-stage techniques. We reviewed the early results of the transanal Soave pullthrough from 6 of the original centers to use it. METHODS: The clinical course of all children with HD undergoing a 1-stage transanal Soave pullthrough between 1995 and 2002 were reviewed. Children with a preliminary stoma or total colonic disease were excluded. RESULTS: There were 141 patients. Mean time between diagnosis and surgery was 32 days, and mean age at surgery was 146 days. Sixty-six (47%) underwent surgery in the first month of life. Forty-seven (33%) had the pathologic transition zone documented laparoscopically or through a small umbilical incision before beginning the anal dissection. Mean blood loss was 16 mL, and no patients required transfusion. Mean time to full feeding was 36 hours, mean postoperative hospital stay was 3.4 days, and 87 patients (62%) required only acetaminophen for pain. Early postoperative complications included perianal excoriation (11%), enterocolitis (6%), and stricture (4%). One patient died of congenital cardiac disease. Mean follow-up was 20 months; 81% had normal bowel function for age, 18% had minor problems, and 1% had major problems. Two patients required a second operation (twisted pullthrough, and residual aganglionosis). One patient developed postoperative adhesive bowel obstruction. CONCLUSION: To date, this report represents the largest series of patients undergoing the 1 stage transanal Soave pullthrough. This approach is safe, permits early feeding, causes minimal pain, facilitates early discharge, and presents a low rate of complications. PMID- 14530729 TI - Left ventricular assist devices as permanent heart failure therapy: the price of progress. AB - BACKGROUND DATA: The REMATCH trial evaluated the efficacy and safety of long-term left ventricular assist device (LVAD) support in stage D chronic end-stage heart failure patients. Compared with optimal medical management, LVAD implantation significantly improved the survival and quality of life of these terminally ill patients. To date, however, there have been no analyses of the cost related to the LVAD survival benefit. This paper addresses the cost of hospital resource use, and its predictors, for long-term LVAD patients. METHODS: Detailed cost data were available for 52 of 68 REMATCH patients randomized to LVAD therapy. We combined the clinical dataset with Medicare data, standard billing forms (UB-92), and line item bills provided directly by clinical centers. Charges were converted to costs by using the Ratio-of-Cost-to-Charges for each major resource category. RESULTS: The mean cost for the initial implant-related hospitalization was $210,187 +/- 193,295. When implantation hospitalization costs are compared between hospital survivors and nonsurvivors, the mean costs increase from $159,271 +/- 106,423 to $315,015 +/- 278,713. Sepsis, pump housing infection, and perioperative bleeding are the major drivers of implantation cost, established by regression modeling. In the patients who survived the procedure (n = 35), bypass time, perioperative bleeding, and late bleeding were the drivers of cost. The average annual readmission cost per patient for the overall cohort was $105,326. CONCLUSIONS: The cost of long-term LVAD implantation is commensurate with other life-saving organ transplantation procedures like liver transplantation. As an evolving technology, there are a number of opportunities for improvement that will likely reduce costs in the future. PMID- 14530731 TI - Training in trauma surgery: quantitative and qualitative aspects of a new paradigm for fellowship. AB - OBJECTIVE: To describe outcomes from a clinical trauma surgical education program that places the board-eligible/board-certified fellow in the role of the attending surgeon (fellow-in-exception [FIE]) during the latter half of a 2-year trauma/surgical critical care fellowship. SUMMARY BACKGROUND DATA: National discussions have begun to explore the question of optimal methods for postresidency training in surgery. Few objective studies are available to evaluate current training models. METHODS: We analyzed provider-specific data from both our trauma registry and performance improvement (PI) databases. In addition, we performed TRISS analysis when all data were available. Registry and PI data were analyzed as 2 groups (faculty trauma surgeons and FIEs) to determine experience, safety, and trends in errors. We also surveyed graduate fellows using a questionnaire that evaluated perceptions of training and experience on a 6 point Likert scale. RESULTS: During a 4-year period 7,769 trauma patients were evaluated, of which 46.3% met criteria to be submitted to the PA Trauma Outcome Study (PTOS, ie, more severe injury). The faculty group saw 5,885 patients (2,720 PTOS); the FIE group saw 1,884 patients (879 PTOS). The groups were similar in respect to mechanism of injury (74% blunt; 26% penetrating both groups) and injury severity (mean ISS faculty 10.0; FIEs 9.5). When indexed to patient contacts, FIEs did more operations than the faculty group (28.4% versus 25.6%; P < 0.05). Death rates were similar between groups (faculty 10.5%; FIEs 10.0%). Analysis of deaths using PI and TRISS data failed to demonstrate differences between the groups. Analysis of provider-specific errors demonstrated a slightly higher rate for FIEs when compared with faculty when indexed to PTOS cases (4.1% versus 2.1%; P < 0.01). For both groups, errors in management were more common than errors in technique. Twenty-one (91%) of twenty-three surveys were returned. Fellows' feelings of preparedness to manage complex trauma patients improved during the fellowship (mean 3.2 prior to fellowship versus 4.5 after first year versus 5.8 after FIE year; P < 0.05 by ANOVA). Eighty percent rated the FIE educational experience "great -5" or "exceptional- 6." Eighty-five percent consider the current structure of the fellowship (with FIE year) as ideal. Ninety percent would repeat the fellowship. CONCLUSION: The educational experience and training improvement offered by the inclusion of a FIE period during a trauma fellowship is exceptional. Patient outcomes are unchanged. The potential for an increased error rate is present during this period of clinical autonomy and must be addressed when designing the methods of supervision of care to assure concurrent senior staff review. PMID- 14530730 TI - Endovascular stent graft repair of abdominal and thoracic aortic aneurysms: a ten year experience with 817 patients. AB - OBJECTIVE: On November 23, 1992, the first endovascular stent graft (ESG) repair of an aortic aneurysm was performed in North America. Following the treatment of this patient, we have continued to evaluate ESG over the past 10 years in the treatment of 817 patients. SUMMARY AND BACKGROUND DATA: Abdominal (AAA) or thoracic (TAA) aortic aneurysms are a significant health concern traditionally treated by open surgical repair. ESG therapy may offer protection from aneurysm rupture with a reduction in procedure morbidity and mortality. METHODS: Over a 10 year period, 817 patients were treated with ESGs for AAA (723) or TAA (94). Patients received 1 of 12 different stent graft devices. Technical and clinical success of ESGs was reviewed, and the incidence of procedure-related complications was analyzed. RESULTS: The mean age was 74.3 years (range, 25-95 years); 678 patients (83%) were men; 86% had 2 or more comorbid medical illnesses, 67% of which included coronary artery disease. Technical success, on an intent-to-treat basis was achieved in 93.8% of patients. Primary clinical success, which included freedom from aneurysm-related death, type I or III endoleak, graft infection or thrombosis, rupture, or conversion to open repair was 65 +/- 6% at 8 years. Of great importance, freedom from aneurysm rupture after ESG insertion was 98 +/- 1% at 9 years. There was a 2.3% incidence of perioperative mortality. One hundred seventy five patients died of causes not related to their aneurysm during a mean follow-up of 15.4 months. CONCLUSIONS: Stent graft therapy for aortic aneurysms is a valuable alternative to open aortic repair, especially in older sicker patients with large aneurysms. Continued device improvements coupled with an enhanced understanding of the important role of aortic pathology in determining therapeutic success will eventually permit ESGs to be a more durable treatment of aortic aneurysms. PMID- 14530732 TI - Quantification of risk for pouch failure after ileal pouch anal anastomosis surgery. AB - OBJECTIVE: To identify risk factors associated with ileal pouch failure and to develop a multifactorial model for quantifying the risk of failure in individual patients. SUMMARY BACKGROUND DATA Ileal pouch anal anastomosis (IPAA) has become the treatment choice for most patients with ulcerative colitis and familial adenomatous polyposis who require surgery. At present, there are no published studies that investigate collectively the interrelation of factors related to ileal pouch failure, nor are there any predictive indices for risk stratification of patients undergoing IPAA surgery. METHODS: Data from 23 preoperative, 7 intraoperative, and 10 postoperative risk factors were recorded from 1,965 patients undergoing restorative proctocolectomy in a single center between 1983 and 2001. Primary end point was ileal pouch failure during the follow-up period of up to 19 years. The "CCF ileal pouch failure" model was developed using a parametric survival analysis and a 70%:30% split-sample validation technique for model training and testing. RESULTS: The median patient follow-up was 4.1 year (range, 0-19 years). Five-year ileal pouch survival was 95.6% (95% CI, 94.4 96.7). The following risk factors were found to be independent predictors of pouch survival and were used in the final multivariate model: patient diagnosis, prior anal pathology, abnormal anal manometry, patient comorbidity, pouch perineal or pouch-vaginal fistulae, pelvic sepsis, anastomotic stricture and separation. The model accurately predicted the risk of ileal pouch failure with adequate calibration statistics (Hosmer Lemeshow chi2 = 3.001; P = 0.557) and an area under the receiver operating characteristics curve of 82.0%. CONCLUSIONS: The CCF ileal pouch failure model is a simple and accurate way of predicting the risk of ileal pouch failure in clinical practice on a longitudinal basis. It may play an important role in providing risk estimates for patients wishing to make informed choices on the type of treatment offered to them. PMID- 14530733 TI - The duodenal switch operation for the treatment of morbid obesity. AB - OBJECTIVE: To determine the safety and efficacy of the duodenal switch procedure as surgical treatment of morbid obesity. SUMMARY BACKGROUND DATA: The longitudinal gastrectomy and duodenal switch procedure as performed for morbid obesity involves a 75% subtotal greater curvature gastrectomy and long limb suprapapillary Roux-en-Y duodenoenterostomy. This results in a restricted caloric intake and diversion of bile and pancreatic secretions to induce fat malabsorption. Broad acceptance of this procedure has been impeded because of concerns that the malabsorptive component may produce serious nutritional complications. METHODS: Review of data collected prospectively from all patients who underwent duodenal switch as the primary surgical treatment of morbid obesity at a single institution during the 10-year period beginning September 1992. Operative morbidity and mortality, weight loss, volume of food intake, and bowel function were recorded. Sequential measurements of serum albumin, hemoglobin, and calcium levels were obtained to assess metabolic function and nutrient absorption. RESULTS: Duodenal switch was performed as the primary operation in 701 (81%) of a total 863 patients undergoing bariatric surgery during the period of study. The average body mass index (BMI) was 52.8 (range, 34-95). Perioperative mortality was 1.4%, and morbidity (including leaks, wound dehiscence, splenectomy, and postoperative hemorrhage) occurred in 21 patients (2.9%). Weight loss averaged 127 pounds at 1 year, 131 at 3 years, and 118 at 5 or more years (% EBWL of 69%, 73%, and 66%, respectively). The mean number of bowel movements was fewer than 3 per day. Patients reported and maintained a mean restriction of 63% of their preoperative intake (approximately 1600 calories), with no specific food intolerance, at 3 or more years follow-up. At 3 years, serum albumin remained at normal levels in 98% of patients, hemoglobin in 52%, and calcium in 71%. No patients reported dumping, and marginal ulcers were not seen. CONCLUSIONS: The longitudinal gastrectomy with duodenal switch is a safe and effective primary procedure for the treatment of morbid obesity. It has the advantage of allowing acceptable alimentation with a minimum of side effects while producing and maintaining significant weight loss. These results are achieved without developing significant dietary restrictions or clinical metabolic or nutritional complications. PMID- 14530734 TI - Influence of surgical subspecialty training on in-hospital mortality for gastrectomy and colectomy patients. AB - OBJECTIVE: This study examined the relationship of surgeon subspecialty training and interests to in-hospital mortality while controlling for both hospital and surgeon volume. SUMMARY BACKGROUND DATA: The relationship between volume of surgical procedures and in-hospital mortality has been studied and shows an inverse relationship. METHODS: A large Statewide Planning and Research Cooperative System was used to identify all 55,016 inpatients who underwent gastrectomy (n = 6434) or colectomy (n = 48,582) between January 1, 1998 and December 31, 2001. Surgical subspecialty training and interest was defined as surgeons who were members of the Society of Surgical Oncology (training/interest; n = 68) or the Society of Colorectal Surgery (training; n = 61) during the study period. The association of in-hospital mortality and subspecialty training/interest was examined using a logistic regression model, adjusting for demographics, comorbidities, insurance status, and hospital and surgeon volume. RESULTS: Overall mortality for colectomy patients was 4.6%; the adjusted mortality rate for subspecialty versus nonsubspecialty-trained surgeons was 2.4% versus 4.8%, respectively (adjusted odds ratio [OR] = 0.45; 95% confidence interval [CI] = 0.34, 0.60; P < 0.0001). Gastrectomy patients experienced an overall mortality rate of 8.4%; the adjusted mortality rate for patients treated by subspecialty trained surgeons was 6.5%, while the adjusted mortality rate for nonsubspecialty trained surgeons was 8.7% (adjusted OR = 0.70; 95% CI = 0.46, 1.08; P = 0.10). CONCLUSIONS: For gastrectomies and colectomies, risk-adjusted mortality is substantially lower when performed by subspecialty interested and trained surgeons, even after accounting for hospital and surgeon volume and patient characteristics. These findings may have implications for surgical training programs and for regionalization of complex surgical procedures. PMID- 14530735 TI - High-resolution source imaging in mesiotemporal lobe epilepsy: a comparison between MEG and simultaneous EEG. AB - Magnetic source imaging is claimed to have a high accuracy in epileptic focus localization and may be a guide for epilepsy surgery. Non-lesional mesiotemporal lobe epilepsy (MTLE), the most common form of epilepsy operated on, has different etiologies, which may affect the choice of surgical approach. The authors compared whole-head magnetoencephalography (MEG) with high-resolution EEG for source identification in MTLE. Nineteen patients with unilateral, nonlesional MTLE underwent a simultaneous 151-channel CTF MEG (CTF Systems, Inc., Port Coquitlam, British Columbia, Canada) and 64-channel EEG recordings with sleep induction. Three independent observers selected spikes from the EEG and MEG recordings separately. Only when there was interobserver agreement (kappa>0.4) on the presence of spikes in recordings were consensus spikes averaged. EEG and MEG equivalent current dipoles (ECD) were then integrated in the head model of the patient reconstructed from MRI. The results were compared with intraoperative electrocorticography findings. Spikes were detected in 32% of MEGs and 42% of EEGs. No patient showed MEG spikes only. Equivalent current dipole modeling correctly localized the source to the temporal lobe in four out of five MEG and three out of eight EEG recordings. MEG localized sources were more superficial and EEG localized sources were deeper. Unfortunately, basal temporal lobe areas were only partially covered by the sensor helmet of the MEG setup. Best correlation between EEG or MEG findings and electrocorticography findings was between horizontal EEG dipole orientation and prominent neocortical spiking; these patients also had a less favorable prognosis. Magnetic source imaging is currently unlikely to alter the surgical management of MTLE. The yield of spikes is too low, and ECD modeling shows only partial correlation with electrocorticography findings. Moreover, the whole-head MEG helmet provides insufficient coverage of the temporal lobe. PMID- 14530736 TI - Continuous electrocorticogram epileptiform discharges due to brain gliosis. AB - Cortical dysplasia is known to produce continuous epileptiform discharges (CEDs) on electrocorticogram (EcoG) and EEG recordings. The authors studied the incidence of CEDs on ECoGs and correlated this data with pathologic findings. Thirty ECoGs were reviewed that were performed on patients with parietal or occipital lobe epilepsy operated on since 1960. CED was classified as: (1) continuous or semicontinuous rhythmic spikes or sharp waves at frequencies ranging from 2 to 8 Hz, and (2) repetitive bursts of rhythmic polyspike activity lasting 2 to 10 s. All nontumoral pathologic specimens were reviewed. Epileptiform activity was classified using the following criteria: focal (one gyrus), regional (two gyri), lobar (three gyri), bilobar, or multilobar. Pathologic examination showed gliosis in eight specimens, focal cortical dysplasia in five specimens, tumoral lesions in eight specimens, and other pathology in nine specimens. CED was found in 11 ECoGs. In seven pathology specimens, significant gliosis was shown, and in the remaining four specimens, a dysplastic lesion was diagnosed. Epileptiform activity was widespread (lobar, bilobar, or multilobar) when gliosis or focal cortical dysplasia was present. Absence of epileptiform activity or a focal/regional distribution was found in tumors and other lesions. These data suggest that extensive gliotic lesions are highly epileptogenic and produce CEDs, which are morphologically undistinguishable from those produced by focal cortical dysplasia. PMID- 14530737 TI - Extent of ictal origin in mesial temporal sclerosis patients monitored with subdural intracranial electrodes predicts outcome. AB - In patients with mesiotemporal sclerosis, posterior hippocampal involvement at the ictal onset is not associated with an excellent outcome. A study confirmed that ictal onset in the posterior parahippocampal gyrus is associated with a less favorable outcome compared with ictal onset in the anterior parahippocampal gyrus in patients with mesiobasal temporal lobe epilepsy who are undergoing foramen ovale recording. The authors hypothesized that involvement of the two medial contact points of posterior basal temporal subdural (SD) strip at the ictal onset, representing ictal onset in the posterior parahippocampal gyrus, may also adversely influence the surgical outcome. With this objective, the authors assessed the incidence of posterior basal temporal SD strip (the two medial contact points) involvement at the ictal onset in patients with mesiotemporal sclerosis and determined whether presence of this finding influenced surgical outcome. Thirty-six patients with mesiotemporal sclerosis underwent a single SD grid (lateral frontotemporal) and strips (three basal temporal and one orbitosubfrontal) monitoring. Based on the earliest involvement of basal temporal strips (the two medial contact points) during the seizure, patients were classified into (1) anterior and/or middle basal temporal, or (2) posterior basal temporal (with or without involvement of anterior and/or middle basal temporal) ictal onset groups. A temporal lobectomy with adequate resection of the ictal onset zone was performed in all patients. Surgical outcome was based on Engel's classification. Six of 36 (17%) patients were classified into the posterior basal temporal ictal onset group. Only two patients from the posterior basal temporal ictal onset group experienced a good outcome compared with 26 of 30 patients from anterior and/or middle basal temporal ictal onset group (P = 0.01). In patients with mesiotemporal sclerosis who were monitored with SD electrodes, involvement of the two medial contact points of posterior basal temporal strip at the ictal onset (representing ictal onset in the posterior parahippocampal gyrus) occurred in 17% of the patients. These patients might not experience an excellent surgical outcome despite including the ictal onset zone in resection. These findings may be useful in presurgical counseling of patients with mesiotemporal sclerosis who undergo intracranial SD monitoring. PMID- 14530738 TI - Independent component analysis as a tool to eliminate artifacts in EEG: a quantitative study. AB - Independent component analysis (ICA) is a novel technique that calculates independent components from mixed signals. A hypothetical clinical application is to remove artifacts in EEG. The goal of this study was to apply ICA to standard EEG recordings to eliminate well-known artifacts, thus quantifying its efficacy in an objective way. Eighty samples of recordings with spikes and evident artifacts of electrocardiogram (EKG), eye movements, 50-Hz interference, muscle, or electrode artifact were studied. ICA components were calculated using the Joint Approximate Diagonalization of Eigen-matrices (JADE) algorithm. The signal was reconstructed excluding those components related to the artifacts. A normalized correlation coefficient was used as a measure of the changes caused by the suppression of these components. ICA produced an evident clearing-up of signals in all the samples. The morphology and the topography of the spike were very similar before and after the removal of the artifacts. The correlation coefficient showed that the rest of the signal did not change significantly. Two examiners independently looked at the samples to identify the changes in the morphology and location of the discharge and the artifacts. In conclusion, ICA proved to be a useful tool to clean artifacts in short EEG samples, without having the disadvantages associated with the digital filters. The distortion of the interictal activity measured by correlation analysis was minimal. PMID- 14530739 TI - Temporal evolution and prognostic significance of postoperative spikes after selective amygdala-hippocampectomy. AB - The purpose of this study was to quantify the temporal evolution and to determine the prognostic significance of spikes on serial postoperative EEGs after selective amygdala-hippocampectomy. The authors performed postoperative EEGs 4 months, 1 year, and 2 years after surgery in 31 patients with unilateral mesial temporal lobe epilepsy. Spike frequency was determined ipsilateral to the resection (group I, no spikes; group II, 1 to 10 spikes; group III, 11 to 20 spikes; group IV, more than 21 spikes during the 30-minute recording). The temporal evolution of postoperative spike frequency was assessed, and these parameters were correlated with surgical outcome. Twenty-two patients showed spikes on postoperative EEG. Spike frequency decreased over time in 14 patients, while no changes or minimal changes occurred in seven patients, and spike frequency increased in one patient. Nine patients had no spikes. There was no correlation between occurrence, frequency, and temporal evolution of spikes with postoperative seizure control. The authors' results demonstrate a progressive decrease of spike frequency during the postoperative period after selective amygdala-hippocampectomy, although occurrence, frequency, and temporal evolution of postoperative spikes had no implications on surgical outcome. PMID- 14530740 TI - Quantitative EEG monitoring during cerebral air embolism and hyperbaric oxygen treatment in a pig model. AB - The purpose of this study was to evaluate the contribution of quantitative EEG (qEEG) to an animal model of cerebral air embolism (CAE). In 12 anesthetized pigs, air was injected into the internal carotid artery, and hyperbaric oxygen (HBO) treatment was started either after 3 minutes or after 60 minutes (United States Navy Treatment Table 6). Off-line spectral analysis was used to determine the frequency content of the EEG signal, and factor analysis was performed to determine the frequency ranges that optimally showed the changes in the power spectrum. Factor analysis revealed two factors that represented different and independent spectral changes during embolization: 0.5 to 7.3 Hz (band 1) and 26.4 to 30.3 Hz (band 2). Shortly after embolization, the power in both bands decreased to a minimum, representing an isoelectric EEG in 11 out of the 12 animals. EEG differences between animals were considerable, despite standardized doses of injected air, and qEEG can objectively assess and quantify these differences in immediate impact of air embolism on brain function. Also, qEEG enabled monitoring of the recovery from the initial embolic event and of the response on treatment. The initial recovery was much more protracted in band 2 than in band 1, but even after completing HBO treatment, qEEG values did not return to baseline values in all animals. In addition, two animals did not survive until the end of the HBO treatment, and qEEG proved to be superior to the other measured hemodynamic variables to detect and ensure a deterioration of brain function. This study showed that qEEG monitoring has significant additional value to monitoring HBO treatment. PMID- 14530741 TI - EEG power and coherence in children with educational problems. AB - SUMMARY: SUMMARY This study deals with the quantitative EEG (QEEG) of children attending schools for the mentally retarded and learning disabled. Questions are in which way do the EEGs of these children differ from normal development and whether deviations are restricted to a subgroup of children. The topographic distribution of EEG power is of particular interest. Based on a sample of n = 158 normal children, age-standardized values of absolute power (delta, theta, alpha 1, alpha 2, beta 1, beta 2 at F4, F3, C4, C3, CZ, PZ, O2, O1) and of coherence are computed for all children. The topographic distribution is assessed by analysis of variance (ANOVA) and by a principal component approach. The EEG of children with educational problems differs substantially from normal development in the slow bands and differs less in the fast bands. Deviations affect a subgroup of children, mainly children attending a school for the mentally retarded. Topographic distribution is an important factor in all bands. Coherence analysis leads to rather weak results that lack a clear interpretation. The QEEG is useful for understanding neurophysiological development in children with educational problems as a group more than individually. Parameters of topographic distribution provide strong additional information to power itself. PMID- 14530742 TI - Patterns of disturbed impulse propagation in multiple sclerosis identified by low and high frequency somatosensory evoked potential components. AB - In human median nerve somatosensory evoked potentials (SSEPs), high frequency (600 Hz) oscillations (HFOs) are superimposed onto the low frequency SSEP component N20. High frequency oscillations are generated both in deep axon segments of thalamo-cortical projection neurons and at the primary somatosensory cortex. The present study aimed to test the hypothesis that HFOs might be more sensitive to temporal dispersion caused by demyelinating lesions in multiple sclerosis (MS) than the N20. The authors recorded HFOs in median nerve SSEPs in 50 patients with definite MS and in 30 healthy controls. Three patterns of SSEP alterations were found: (1) abolished HFOs with either normal (11% of stimulated limbs), or delayed N20 (16% of stimulated limbs); (2) an attenuation of N20 amplitude with preserved HFOs (13%); and (3) a mixture of both patterns (21%). The first pattern--normal N20 with abolished HFOs--indicates that the HFOs are a sensitive marker of slight demyelination. The second pattern is suggestive of a mainly axonal lesion type, while the third pattern points to a combined axonal/demyelinating process or a conduction block. Analysis of HFOs allows identification of slight demyelinating processes in MS patients in whom the N20 SSEP component remains unaffected. The HFOs provide a tool to distinguish different patterns of disturbed impulse propagation. PMID- 14530743 TI - Evaluation and comparison of Tranquility and AutoSet T autotitrating CPAP machines. AB - The objective of this study was to determine the effectiveness of two autotitrating continuous positive airway pressure (auto-CPAP), the Tranquility (Respironics, Inc., Murrysville, PA, U.S.A.) and the AutoSet T (ResMed Corp., Poway, CA, U.S.A). Patients with obstructive sleep apnea syndrome (OSAS) underwent attended auto-CPAP titration with either the Tranquility or AutoSet T machine. The auto-CPAP machine was given a performance score between 0 and 9. A score of 3 or lower indicated that autotitration had to be discontinued. Sixty patients were studied, 35 patients with the Tranquility machine and 25 patients with the AutoSet T machine. The mean age was 55.4 years, whereas the mean body mass index (BMI) was 32.7. The mean respiratory disturbance index (RDI) was 37.5, and the mean arousal index before use of auto-CPAP was 41.4. The two treatment groups were comparable in age, BMI, RDI, and arousal index. The mean performance scores for the Tranquility and AutoSet T machines were not statistically different. Autotitration with the Tranquility machine had to be discontinued in 10 of the 35 patients (29%), whereas with the AutoSet T machine, autotitration was discontinued in 11 of the 25 patients (44%). Despite advances in auto-CPAP machines, there remains a large minority of patients with OSAS in whom these machines do not accomplish treatment goals. At this time, these machines are not a substitute for in-laboratory CPAP titration polysomnograms. PMID- 14530744 TI - Clearing up some inaccuracies about the American Board of Neurophysiologic Monitoring exam for intraoperative monitoring. PMID- 14530745 TI - American Board of Neurophysiologic Monitoring intraoperative monitoring credentialing is for physicians, too. PMID- 14530746 TI - Cell therapy with a tissue-engineered kidney reduces the multiple-organ consequences of septic shock. AB - OBJECTIVE: Gram-negative septic shock has a clinical mortality rate approaching 50%. The cause of death is secondary to a systemic inflammatory response syndrome with resulting cardiovascular collapse, ischemic damage to vital organs, and multiple-organ systems failure. Renal tubule cell injury occurs early in septic shock but is not clinically appreciated. Since renal tubule cells appear to play a critical role in the immunoregulation of stress states, renal cell therapy during septic shock may alter the detrimental multiple-organ consequences of systemic Gram-negative infection. The development of a tissue-engineered bioartificial kidney consisting of a conventional hemofiltration cartridge in series with a renal tubule assist device (RAD) containing 109 renal proximal tubule cells may be a new therapeutic approach to this clinical disorder. DESIGN: Laboratory study. SETTING: University medical school. SUBJECTS: Pigs weighing 30 35 kg. INTERVENTIONS: To assess the effect of the bioartificial kidney and the RAD in septic shock, pigs were administered 30 x 10(10) bacteria/kg body weight of Escherichia coli into the peritoneal cavity and within 1 hr were immediately placed in a continuous venovenous hemofiltration extracorporeal circuit with either a sham RAD without cells or a RAD with cells. MEASUREMENTS AND MAIN RESULTS: In this animal model, septic shock resulted within hours in acute tubule necrosis in the kidneys of all animals. Renal cell therapy resulted in significantly higher cardiac outputs and renal blood flow rates in treated animals compared with sham controls. RAD treatment also was associated with significantly lower plasma circulating concentrations of interleukin-6 and interferon-gamma compared with sham-treated animals. IL-6 release rates from peripheral blood mononuclear cells isolated from RAD-treated animals were significantly higher after endotoxin stimulation than those isolated from control animals. These physiologic and molecular alterations were associated with nearly a doubling of the average survival time in the RAD-treated group compared with the sham control group. CONCLUSION: These results demonstrate that renal cell therapy ameliorates cardiac and vascular dysfunction, alters systemic cytokine abnormalities, and improves survival time in a large animal model of Gram negative septic shock. A cell therapeutic approach with a tissue-engineered bioartificial kidney may be a new treatment modality for this current unmet medical need. PMID- 14530747 TI - Parenteral administration of glipizide sodium salt, an inhibitor of adenosine triphosphate-sensitive potassium channels, prolongs short-term survival after severe controlled hemorrhage in rats. AB - OBJECTIVE: Recent experimental evidence suggests that activation of adenosine triphosphate (ATP)-sensitive potassium channels contributes to vascular failure and early mortality after hemorrhagic shock. The present investigation evaluated the effects of the water-soluble sodium salt of glipizide, an inhibitor of ATP sensitive potassium channels, in anesthetized and awake rats subjected to severe controlled hemorrhage. DESIGN: Prospective, randomized, controlled study. SETTING: Animal research laboratory. SUBJECTS: Male Wistar rats. INTERVENTIONS: Anesthetized rats were subjected to bleeding to reduce mean arterial pressure to either 40 or 35 mm Hg, which was maintained constant for 60 mins. In addition, awake rats underwent blood withdrawal of 4.25 mL/100 g over 20 mins. At the end of the hemorrhage period and 30 mins later, the animals received intravenous (5 and 20 mg/kg) or intramuscular (10 and 40 mg/kg) injections of glipizide sodium salt or vehicle. MEASUREMENTS AND MAIN RESULTS: In anesthetized rats subjected to pressure-controlled hemorrhage, glipizide sodium salt improved mean arterial pressure in a dose-dependent manner. Compared with the vehicle-treated animals, mean arterial pressure increased from 41.6 +/- 4.6 to 63.1 +/- 3.1 mm Hg in the 20 mg/kg intravenous group and from 33.2 +/- 4.9 to 54.0 +/- 4.7 mm Hg in the 40 mg/kg intramuscular group 60 mins after a 40-mm Hg shock. Furthermore, the drug did not affect the hemorrhage-induced changes in blood glucose concentrations. However, the higher doses of glipizide sodium salt attenuated the increments in plasma concentrations of lactate, alanine aminotransferase, creatinine, and amylase. Moreover, the higher doses markedly improved short-term survival after pressure- and volume-controlled bleeding. Overall, the intramuscular injections of the drug exerted salutary effects that were comparable to the intravenous administration. CONCLUSIONS: In rats, parenteral administration of the water soluble glipizide sodium salt attenuates vascular and end-organ dysfunction associated with severe hemorrhagic shock and prolongs short-term survival. The intramuscular administration provides comparable benefits as obtained by the intravenous injection. PMID- 14530748 TI - Use of a fully simulated intensive care unit environment for critical event management training for internal medicine residents. PMID- 14530749 TI - Decreased mortality and infectious morbidity in adult burn patients given enteral glutamine supplements: a prospective, controlled, randomized clinical trial. AB - OBJECTIVE: Enteral glutamine supplements have been shown to reduce infectious morbidity in trauma patients, but their effect on burn patients is not known. The objective of this study was to measure the impact of enteral glutamine supplementation on infectious morbidity, length of care, and the immune system in burn patients. DESIGN: Double-blinded, randomized clinical trial. SETTING: Burn center. PATIENTS: Forty-five adults with severe burns. INTERVENTIONS: Patients were randomized to receive either glutamine or an isonitrogenous control mixture until complete healing occurred. Length of care, incidence of positive blood culture, and mortality were recorded. Phagocytosis by circulating polymorphonuclear cells was measured every 3 days. MEASUREMENTS AND MAIN RESULTS: Patient characteristics were similar in both groups. Four patients were excluded from the analysis, because three of them died within 72 hrs and the fourth could not receive enteral nutrition and amino acid supplements for the first 10 days. Of the remaining 41 patients, length of care in the survivors was not different between groups (0.9 vs. 1.0 days/percent total body surface area for glutamine vs. control, respectively), positive blood culture was three times more frequent in control than in glutamine treatment (4.3 vs. 1.2 days/patient, p <.05), and Pseudomonas aeruginosa was detected in six patients on control and zero on glutamine (p <.05). Phagocytosis by polymorphonuclear cells was not different between groups. Mortality rate was significantly lower in glutamine than in control: intention to treat, two vs. 12 (p <.05); per protocol analysis, zero vs. eight (p <.01). CONCLUSIONS: Enteral glutamine supplementation in adult burn patients reduces blood infection by a factor of three, prevents bacteremia with P. aeruginosa, and may decrease mortality rate. It has no effect on level of consciousness and does not appear to influence phagocytosis by circulating polymorphonuclear cells. PMID- 14530750 TI - Citrate pharmacokinetics and metabolism in cirrhotic and noncirrhotic critically ill patients. AB - OBJECTIVES: To investigate pharmacokinetics and metabolism of sodium citrate in critically ill patients. To determine the risk of citrate accumulation in the setting of liver dysfunction (cirrhosis, hepatorenal syndrome). DESIGN: Prospective cohort study. SETTING: Intensive Care Unit, Department of Medicine IV, University Hospital Vienna. PATIENTS: Consecutive critically ill cirrhotic (n = 16) and noncirrhotic patients (n = 16). INTERVENTIONS: Infusion of sodium citrate (0.5 mmol.kg-1.hr-1) and calcium chloride (0.17 mmol.kg-1.hr-1) for 2 hrs. Analysis of serial arterial blood samples. MEASUREMENTS AND MAIN RESULTS: Total body clearance of citrate was normal in noncirrhotic critically ill patients but significantly reduced in cirrhotic patients (710 vs. 340 mL/min, p =.008). Citrate peak concentrations and concentration over time were increased by 65% and 114% in cirrhotic patients (p <.001), respectively; volumes of distribution were similar. Net metabolic changes were quantitatively similar, with pH and plasma bicarbonate concentrations increasing more slowly in cirrhotic patients. No citrate-related side effects were noted. Citrate clearance could not be predicted by standard liver function tests and was not appreciably influenced by renal function and Acute Physiology and Chronic Health Evaluation II scores. CONCLUSIONS: This first systematic study on citrate pharmacokinetics and metabolism in critically ill patients confirms a major role of hepatic citrate metabolism by demonstrating reduced citrate clearance in cirrhotic patients. Pharmacokinetic data could provide a basis for the clinical use of citrate anticoagulation in critically ill patients. Provided dose adaptation and monitoring of ionized calcium, citrate anticoagulation seems feasible even in patients with decompensated cirrhosis. Metabolic consequences of citrate infusion were not different between groups in this study but may be more pronounced in prolonged infusion. PMID- 14530751 TI - Rehabilitation after critical illness: a randomized, controlled trial. AB - OBJECTIVE: To evaluate the effectiveness of a rehabilitation program following critical illness to aid physical and psychological recovery. DESIGN: Randomized controlled trial, blind at follow-up with final assessment at 6 months. SETTING: Two district general hospitals and one teaching hospital. PATIENTS: Patients were 126 consecutively admitted intensive care patients meeting the inclusion criteria. INTERVENTIONS: Control patients received ward visits, three telephone calls at home, and clinic appointments at 8 wks and 6 months. Intervention patients received the same plus a 6-wk self-help rehabilitation manual. MEASUREMENTS AND MAIN RESULTS: We measured levels of depression and anxiety (Hospital Anxiety and Depression Scale), phobic symptoms (Fear Index), posttraumatic stress disorder (PTSD)-related symptoms (Impact of Events Scale), and scores on the Short-Form Health Survey physical dimension 8 wks and 6 months after intensive care unit (ICU) treatment. Memory for ICU was assessed at 2 wks post-ICU discharge using the ICU Memory Tool.The intervention group improved, compared with the control patients, on the Short-Form Health Survey physical function scores at 8 wks and 6 months (p =.006), and there was a trend to a lower rate of depression at 8 wks (12% vs. 25%). However, there were no differences in levels of anxiety and PTSD-related symptoms between the groups. The presence of delusional memories was correlated significantly with both anxiety and Impact of Events Scale scores. CONCLUSIONS: A self-help rehabilitation manual is effective in aiding physical recovery and reducing depression. However, in those patients recalling delusional memories from the ICU, further psychological care may be needed to reduce the incidence of anxiety and PTSD-related symptoms. PMID- 14530752 TI - Effect of granulocyte-macrophage colony-stimulating factor on the immune response of circulating monocytes after severe trauma. AB - OBJECTIVE: Severe injury compromises functions of the antigen-presenting immune cells, resulting in an increased vulnerability toward bacterial sepsis. Support of the immune capabilities contributes a desirable therapeutic option in high risk patients. Factors possessing immunostimulating properties such as granulocyte-macrophage colony-stimulating factor (GM-CSF) may serve as potential tools to compensate immunosuppression caused by severe trauma. In the present study, therefore, GM-CSF was examined with regard to its capacity to overcome trauma-induced down-regulation of immune functions. DESIGN: Prospective clinical experimental study. SETTING: University hospital intensive care unit and research facility. PATIENTS: Severely injured patients with >25 points on the Injury Severity Score. INTERVENTIONS: Blood samples of severely injured patients were incubated in vitro with 10 ng/mL GM-CSF for 6 hrs. MEASUREMENTS: Human leukocyte antigen (HLA)-DR expression on monocytes was analyzed by flow cytometry, lipopolysaccharide-induced tumor necrosis factor (TNF)alpha and interleukin-10 production of blood samples was measured by means of enzyme-linked immunoabsorbent assay. MAIN RESULTS: Compared with blood specimens of healthy donors, ex vivo endotoxin-induced TNF alpha production and HLA-DR expression on monocytes were significantly reduced in blood of trauma patients. Ex vivo treatment of blood specimens with GM-CSF increased HLA-DR expression and TNF alpha production stimulated by lipopolysaccharides in both healthy volunteers and patients on day 1 after trauma. Blood samples of patients with an uneventful recovery showed nearly normal TNF alpha synthesis and HLA-DR expression after 2-3 wks, whereas TNF alpha production and HLA-DR expression of patients with sepsis and multiple organ failure remained at low levels. In the sepsis/multiple organ failure group, GM-CSF also enhanced HLA-DR expression and TNF alpha production, although the levels of the volunteers' blood were not reached. CONCLUSIONS: The presented data show that trauma- and sepsis-induced depression of monocyte functions can be counteracted by GM-CSF in vitro, suggesting that this substance may serve as support of immune functions in severely injured patients. PMID- 14530753 TI - Use of xenon as a sedative for patients receiving critical care. AB - OBJECTIVE: Many sedative regimens are used in the intensive care setting, but none are wholly without adverse effect. Xenon is a noble gas with sedative and analgesic properties. It has been used successfully as a general anesthetic and has many desirable properties, not least of which is a minimal effect on the myocardium. In theory, xenon may provide sedation without adverse effect for certain groups of critically ill patients. The objective of this study was to assess the feasibility of using xenon as an intensive care sedative. DESIGN: Double-blind, randomized study. SETTING: Tertiary-level intensive care unit. SUBJECTS: Twenty-one patients admitted to an intensive care unit following elective thoracic surgery. INTERVENTIONS: A standard intensive care sedation regimen (intravenous propofol at 0-5 mg.kg-1.hr-1 and alfentanil 30 microg.kg 1.hr-1) was compared with a xenon sedation regimen delivered using a novel bellows-in-bottle delivery system. MEASUREMENTS AND MAIN RESULTS Each sedative regimen was continued for 8 hrs. The hemodynamic effects, additional analgesic requirements, recovery from sedation, and effect on hematological and biochemical variables were compared for the two sedation regimens. All patients were successfully sedated during the xenon regimen. The mean +/- SD end-tidal xenon concentration required to provide sedation throughout the duration of the study was 28 +/- 9.0% (range, 9-62%). Arterial systolic, diastolic, and mean pressures showed a greater tendency for negative gradients in patients receiving the propofol regimen (p <.05, p <.1, and p <.01, respectively). Recovery following xenon was significantly faster than from the standard sedation regimen (p <.0001). Hematological and biochemical laboratory markers were within normal clinical limits in both groups. CONCLUSIONS: Xenon provided satisfactory sedation in our group of patients. It was well tolerated with minimal hemodynamic effect. Recovery from this agent is extremely rapid. We have demonstrated the feasibility of using xenon within the critical care setting, without adverse effect. PMID- 14530754 TI - Clinical impact of pneumonia caused by Acinetobacter baumannii in intubated patients: a matched cohort study. AB - OBJECTIVE: To determine whether ventilator-associated pneumonia caused by Acinetobacter baumannii (VAPAB) is associated with increased mortality rate. DESIGN: A retrospective matched case-control study in which all intensive care unit adult patients with microbiologically documented VAPAB were defined as cases. SETTING: Four intensive care units from teaching hospitals. PATIENTS: Sixty patients were matched to sixty controls. MEASUREMENTS AND MAIN RESULTS: Controls were matched based on stay before pneumonia onset, disease severity (Acute Physiology and Chronic Health Evaluation II) at admission, and diagnostic category. Population characteristics and intensive care unit mortality rates of patients with VAPAB and their controls were compared. Attributable mortality was determined by subtracting the crude mortality rate of the controls from the crude mortality rate of the case patients. Twenty-four of the 60 case patients died, representing a crude mortality rate of 40%, whereas 17 of the 60 controls died, a crude mortality rate of 28.3% (p =.17). Crude intensive care unit mortality was the same (12 of 35, 34.2%) in patients with VAPAB caused by strains sensitive to imipenem and in their matched controls. It was 44% for the 25 patients with imipenem-resistant strains with an estimated attributable mortality rate of 20.0% (95% confidence interval, -5.6% to 45.7%). Mean intensive care unit stay of patients and controls was 35.3 and 36.6 days, respectively (p = nonsignificant). CONCLUSION: In intubated patients, pneumonia by A. baumannii is not significantly associated with attributable mortality rate or an increased length of intensive care unit stay. PMID- 14530755 TI - Discrepancies between ordered and delivered concentrations of opiate infusions in critical care. AB - OBJECTIVES: We sought to test the assumption that the measured concentrations of medication infusions are within pharmaceutical standards (+/-10% of intended concentrations) and whether, at the time the infusion was mixed, the professional background of persons preparing the infusion or the unit for which the infusion was prepared were related to the observed variation. DESIGN, SETTING, AND PARTICIPANTS: This prospective, observational study was conducted in the neonatal and pediatric intensive care units of a university-affiliated tertiary pediatric center. Morphine infusions prepared for clinical use were randomly sampled over a 7-month period. Those with no error between labeled and ordered concentration were further analyzed. High-performance liquid chromatography was used to determine the concentration of morphine infusions. The primary outcome was a difference of >10% between ordered and measured concentrations. MEASUREMENTS AND MAIN RESULTS: The measured concentration of 65% of the 232 infusions was >10% different from the ordered concentration (95% confidence interval, 58-71%). The concentrations of 6% of infusions represented two-fold errors (95% confidence interval, 3-9%). The difference was normally distributed around zero, suggesting a cumulative effect of random errors, rather than a systematic bias. The time that the infusion was prepared, the professional background of the persons preparing the infusion, and the unit for which the infusion was mixed were not significant predictors of discrepancy (p =.74, analysis of variance). CONCLUSIONS: The concentration of two thirds of infusions prepared for clinical use was outside accepted industry standards. These findings are likely to be broadly representative of intravenous drug administration in hospitalized children and pediatric pharmacokinetic studies. Further study of the causes and clinical impact is required. PMID- 14530756 TI - Changes in intensive care unit nurse task activity after installation of a third generation intensive care unit information system. AB - OBJECTIVE To determine the percentage of time that intensive care unit (ICU) nurses spend on documentation and other nursing activities before and after installation of a third-generation ICU information system. DESIGN: Prospective data collection using real-time time-motion analysis, before and after installation of the ICU information system. SETTING: A ten-bed surgical ICU at a Veterans Affairs medical center. SUBJECTS: ICU nurses. INTERVENTIONS: Installation of a third-generation ICU information system. MEASUREMENTS AND MAIN RESULTS: Ten ICU nurses were studied before and after installation of the ICU information system. Each ICU nurse's activities and tasks, during 4-hr observation periods, were categorized in real-time by a nurse observer and recorded in a laptop computer. Each recorded task was automatically time-stamped and logged into a data file. The percentage of time spent on documentation decreased from 35.1 +/- 8.3% to 24.2 +/- 7.6% (p =.025) after the ICU information system was installed. The percentage of time providing direct patient care increased from 31.3 +/- 9.2% to 40.1 +/- 11.7% (p =.085). The percentage of time doing patient assessment, a direct patient care task, increased from 4.0 +/- 4.7% to 9.4 +/- 4.4% (p =.001). CONCLUSIONS: Installation of a third-generation ICU information system decreased the percentage of time ICU nurses spent on documentation by >30%. Almost half of the time saved on documentation was spent on patient assessment, a direct patient care task. PMID- 14530757 TI - Effects of hypertonic/hyperoncotic treatment after rat cortical vein occlusion. AB - OBJECTIVE: To examine the effects of hypertonic/hyperoncotic treatment on physiologic variables and regional cerebral blood flow and to test its neuroprotective efficiency in a model of permanent venous ischemia. DESIGN: Randomized prospective study. SETTING: University research institute. SUBJECTS: Adult male Wistar rats, weighing 359 +/- 54 g (n = 38). INTERVENTIONS: Rats were subjected to photochemical occlusion of two adjacent cortical veins. A randomized infusion with vehicle (0.9% NaCl), 10% hydroxyethyl starch 200,000 (HES), or 7.5% saline plus 10% hydroxyethyl starch 200,000 (HHES) was started 30 mins after two vein occlusion. Effects on physiologic variables and regional cerebral blood flow (assessed by laser Doppler flowmetry) were studied up to 120 mins after two-vein occlusion. Two days after occlusion, the brains were removed for histologic evaluation. MEASUREMENTS AND MAIN RESULTS: After occlusion, regional cerebral blood flow decreased by 50%, significantly in all groups (from 47.3 +/- 3 to 22.2 +/- 2.2 laser Doppler units). In the vehicle and HES groups, regional cerebral blood flow further decreased to 12.9 +/- 1.9 and 17.8 +/- 2.3 laser Doppler units, respectively. HHES improved regional cerebral blood flow significantly to 27.3 +/- 3.5 laser Doppler units, particularly by reducing no-flow/low-flow areas and reducing infarct size. CONCLUSION: We found that HHES reduced infarct size as a consequence of an improved regional cerebral blood flow and reduced no-flow/low flow areas in the tissue at risk in the two-vein occlusion model. PMID- 14530758 TI - Nasopharyngeal cooling selectively and rapidly decreases brain temperature and attenuates neuronal damage, even if initiated at the onset of cardiopulmonary resuscitation in rats. AB - OBJECTIVE: To determine the effectiveness of nasopharyngeal cooling for selective brain cooling and neuroprotection from ischemia. DESIGN: Prospective animal study. SETTING: Experimental laboratory in a university hospital. SUBJECTS: Male Wistar rats (n = 28). INTERVENTIONS: In study 1, hippocampal temperature was decreased to 31 degrees C under spontaneous circulation. In the nasopharyngeal cooling group, physiologic saline (5 degrees C) was infused to the bilateral nasal cavities at the rate of 100 mL.min-1.kg weight-1. In the whole body cooling group, a fan and a water blanket (5 degrees C) were used. In study 2, ischemia and resuscitation were performed in normothermic and nasopharyngeal cooling (initiated with resuscitation after 5 mins of ischemia and continued for 20 mins) groups. MEASUREMENTS AND MAIN RESULTS: The hippocampal temperature was decreased to 31 degrees C in 7 +/- 2 mins without any change in the rectal temperature in the nasopharyngeal cooling group, whereas a decrease in hippocampal temperature to 31 degrees C took 33 +/- 1 mins in the whole body cooling group. Although skull base region was cooled by nasopharyngeal cooling, the epidural temperature of the parietal region was lower than the hippocampal temperature, indicating that brain temperature was hematogenously lowered. There was no difference between changes in cerebral blood flow or between the ratios of oxygen extraction from arterial blood in the head region in the nasopharyngeal cooling and whole body cooling groups. In the second study, all animals were successfully resuscitated, and the times required for recovery of mean arterial blood pressure (60 mm Hg) after resuscitation in the nasopharyngeal cooling and normothermic groups were the same. The histologic damage was significantly attenuated in the nasopharyngeal cooling group (33 +/- 21% cell death in the hippocampus) compared with that in the normothermic group (73 +/- 11%). CONCLUSIONS: Nasopharyngeal cooling enables rapid and selective reductions in cortical and subcortical temperatures without disturbing the recovery of systemic circulation after resuscitation. PMID- 14530759 TI - Intrarenal blood flow distribution in hyperdynamic septic shock: Effect of norepinephrine. AB - OBJECTIVES: To measure changes in medullary and cortical renal blood flow during experimental hyperdynamic sepsis and the effect of subsequent norepinephrine infusion on such flows. DESIGN Experimental animal study. SETTING: Animal laboratory of university-affiliated physiology institute. SUBJECTS Eighteen anesthetized merino sheep. INTERVENTIONS: A transit-time flow probe was placed around the left renal artery. Laser Doppler flow probes were inserted in the left renal medulla and cortex by micromanipulation to measure changes in regional intrarenal blood flow. MEASUREMENTS AND MAIN RESULTS: Systemic pressures, cardiac output, renal, and intrarenal blood flows were measured continuously. A bolus of Escherichia coli (7.5 x 10(9) colony forming units) was given intravenously to induce hyperdynamic sepsis. After the onset of hyperdynamic sepsis, all animals were randomly allocated to either norepinephrine (0.4 microg.kg-1.min-1 for 30 mins) or observation for 30 mins in random order. E. coli injection induced a significant decrease in mean arterial pressure (102.2 +/- 15.2 mm Hg to 74.3 +/- 16.1 mm Hg, p <.05) and an increase in mean cardiac output (4.60 +/- 1.62 L/min to 5.93 +/- 1.18 L/min, p <.05). However, renal blood flow did not change significantly (326.4 +/- 139.4 mL/min to 293.1 +/- 117.5 mL/min, not significant) despite a 30% increase in renal conductance (3.27 +/- 1.52 to 4.13 +/- 2.01 mL.min-1.mm Hg-1, p <.05). Cortical blood flow decreased by 15% (not significant) and medullary flow by 5% (not significant) during sepsis, but individual changes were unpredictable. On the other hand, norepinephrine infusion caused a significant improvement in mean arterial pressure (74.3 +/- 16.1 to 105.7 +/- 17.7 mm Hg, p <.05) and a further increase in cardiac output (5.93 +/- 1.18 to 7.13 +/- 1.52 L/min, p <.05). Mean renal blood flow also increased (293.1 +/- 117.5 to 384.5 +/- 168.1 mL/min, p <.05) despite decreased renal conductance (4.13 +/- 2.01 to 3.73 +/- 1.91 mL.min-1.mm Hg-1, p <.05). Infusion of norepinephrine significantly increased medullary blood flow by 35% compared with baseline (p <.05) and by 54% compared with untreated sepsis (p <.05), whereas the increases in cortical blood flow (16 and 53%, respectively) were not significant. CONCLUSIONS: Hyperdynamic sepsis caused renal vasodilation but had limited effects on regional intrarenal blood flow. Norepinephrine infusion (0.4 microg.kg 1.min-1) during sepsis significantly increased global and medullary renal blood flow and restored renal vascular tone toward but not above normal. PMID- 14530760 TI - Application of continuous positive airway pressure to trace static pressure volume curves of the respiratory system. AB - OBJECTIVE: To evaluate a new technique for pressure-volume curve tracing. DESIGN: Prospective experimental study. SETTING: Animal research laboratory. SUBJECTS: Six anesthetized rats. INTERVENTIONS: Two pressure-volume curves were obtained by means of the super-syringe method (gold standard) and the continuous positive airway pressure (CPAP) method. For the CPAP method, the ventilator was switched to CPAP and the pressure level was raised from 0 to 50 cm H2O in 5 cm H2O steps and then decreased, while we measured lung volume using respiratory inductive plethysmography. Thereafter, lung injury was induced using very high-volume ventilation. Following injury, two further pressure-volume curves were traced. Pressure-volume pairs were fitted to a mathematical model. MEASUREMENTS AND MAIN RESULTS: Pressure-volume curves were equivalent for each method, with intraclass correlation coefficients being higher than.75 for each pressure level measured. Bias and precision for volume values were 0.46 +/- 0.875 mL in basal measurements and 0.31 +/- 0.67 mL in postinjury conditions. Lower and upper inflection points on the inspiratory limb and maximum curvature point on the deflation limb obtained using both methods and measured by regression analysis also were correlated, with intraclass correlation coefficients (95% confidence interval) being.97 (.58,.99),.85 (.55,.95), and.94 (.81,.98) (p <.001 for each one). When inflection points were estimated by observers, the correlation coefficient between methods was.90 (.67,.98) for lower inflection points (p <.001). However, estimations for upper inflection points and maximum curvature point were significantly different. CONCLUSIONS: The CPAP method for tracing pressure-volume curves is equivalent to the super-syringe method. It is easily applicable at the bedside, avoids disconnection from the ventilator, and can be used to obtain both the inspiratory and the deflation limbs of the pressure-volume curve. Use of regression techniques improves determination of inflection points. PMID- 14530761 TI - High-frequency oscillation and exogenous surfactant administration in lung injured adult sheep. AB - OBJECTIVE: To evaluate the effects of high-frequency oscillation on the response to exogenous surfactant in lung-injured adult sheep. DESIGN: A prospective, controlled, in vivo, animal laboratory study. SETTING: Animal research facility of a health sciences university. SUBJECTS: Twenty-eight adult sheep. INTERVENTIONS: Animals were anesthetized and instrumented with a tracheostomy and vascular catheters. Following whole lung saline lavage, animals were randomized to one of four groups: Group S-CMV received surfactant and was ventilated for 4 hrs using a conventional mechanical ventilation strategy, group S-HFOV/CMV received surfactant and was ventilated with a high-frequency oscillation technique for 2 hrs and a conventional mechanical strategy for 2 hrs, group HFOV/CMV underwent the latter ventilatory strategies without receiving surfactant, and group HFOV was ventilated with high-frequency oscillation only for 4 hrs. At the end of the ventilatory period, the distributions of ventilation and surfactant were evaluated in animals that received surfactant. MEASUREMENTS AND MAIN RESULTS: Animals in the S-CMV group had a significantly greater mean PaO2 value at the end of the experimental period than animals in the S-HFOV/CMV or HFOV/CMV groups. Evaluation of the distribution of ventilation relative to surfactant demonstrated that animals ventilated with high-frequency oscillation followed by conventional mechanical ventilation had a significantly greater disproportionate distribution of ventilation relative to surfactant compared with the CMV-only group. CONCLUSIONS: A period of high-frequency oscillation, as used in this study, immediately following exogenous surfactant administration mitigates the host's response to surfactant when subsequently switched to conventional mechanical ventilation. PMID- 14530762 TI - Increased expression of 19-kD interacting protein-3-like protein and the relationship to apoptosis in the lung of rats with severe acute pancreatitis. AB - OBJECTIVE: The aim of the present study was to determine the underlying cellular mechanisms in the pancreas after acute pancreatitis and to study the pathogenesis of pancreatitis-associated lung injury. We applied a differential display analysis to normal pancreas and to the pancreas with acute pancreatitis in rats, and we examined the expression of the identified gene in the lung as well as the pancreas after acute pancreatitis. DESIGN: Controlled animal study. SETTING: Research laboratory of an academic institution. SUBJECTS: Ninety male Wistar rats. INVESTIGATIONS: Pancreatitis was induced by retrograde intraductal infusion of 4% sodium taurocholate (100 microL/100 g of body weight). Data were compared with data from controls (sham). MEASUREMENTS AND MAIN RESULTS: We cloned some expressed sequence tags and identified one complementary DNA fragment. The deduced protein was a polypeptide of 218 amino acids, which was almost identical to human 19-kD interacting protein-3-like (NIP3L) protein. The expression of rat NIP3L identified in this study increased slightly in the pancreas after induction of acute pancreatitis but showed a marked increase in the lung by both Northern and Western blot analysis. NIP3L immunoreactivity was noted in alveolar and epithelial cells of the control (sham) lung, and the immunoreactivity in these cells was elevated after induction of acute pancreatitis. Moreover, acute pancreatitis increased terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling-positive alveolar and bronchiolar cells in the lung. CONCLUSION: NIP3L may be involved in lung injury, which is one of the major causes of death in cases of severe acute pancreatitis. PMID- 14530763 TI - Decompressive craniectomy for severe traumatic brain injury: Evaluation of the effects at one year. AB - OBJECTIVE: To assess the effect on outcome (1 yr) of decompressive craniectomy performed within or after the first 24 hrs post-trauma in severely head-injured trauma patients with intractable cerebral hypertension. DESIGN: Retrospective cohort study. SETTINGS: Intensive care unit of a university hospital. PATIENTS: Among 816 patients with severe head trauma (Glasgow Coma Scale < or =8), 40 underwent decompressive craniectomy. After data collection, patients were divided into two groups: early and late decompressive craniectomy. An early decompressive craniectomy was performed within the first 24 hrs in patients according to the following criteria: a Glasgow Coma Scale score <6 and the existence of clinical signs of cerebral herniation (absence of pupillary reflexes), correlated with abnormalities in computed tomography scan including hematoma, appearance of diffuse or unilateral brain swelling, and/or cerebral herniation. The intracranial pressure in these patients was not measured before the decompressive craniectomy was performed. A late decompressive craniectomy (>24 hrs) was performed according to following criteria: an intractable intracranial hypertension with intracranial pressure >35 mm Hg, a unilateral or bilateral absence of pupillary reflexes, and the same abnormalities in computed tomography scan as previously described. INTERVENTION: Twenty-seven patients with signs of cerebral herniation required the procedure at the time of initial evacuation of a mass lesion. In 13 patients, decompressive craniectomy was performed because of elevated intracranial pressure refractory to medical treatment consisting of cerebrospinal fluid derivation, deep sedation, osmotherapy, hyperventilation, and nesdonal or propofol. MEASUREMENTS AND MAIN RESULTS: Five patients (19%) in whom an early decompressive craniectomy was performed had good recoveries (social rehabilitation), eight patients (30%) remained in a persistent vegetative state or with a severe disability, and 14 died (52%). On the other hand, the performance of late decompressive craniectomy in case of medical treatment failure was followed by social rehabilitation in five patients (38%) and death in three patients (23%). A persistent vegetative state or a severe disability was observed in five patients (38%). Meningitis or cerebral abscess occurred in six patients after decompressive craniectomy and were easily cured by antibiotic treatment. CONCLUSIONS: In 40 patients with intractable intracranial hypertension and at very high risk of brain death, decompressive craniectomy allowed 25% of patients to attain social rehabilitation at 1 yr. PMID- 14530764 TI - Capnometry for noninvasive continuous monitoring of metabolic status in pediatric diabetic ketoacidosis. AB - OBJECTIVE: To determine the utility of continuous noninvasive capnometry for monitoring pediatric patients with diabetic ketoacidosis as assessed by the agreement between end-tidal carbon dioxide (PetCO2) and PCO2 DESIGN: Clinical, prospective, observational study. SETTING: University affiliated children's hospital. INTERVENTIONS: Patients with diabetic ketoacidosis were monitored with an oral/nasal carbon dioxide (CO2) sampling cannula while in the emergency department. Laboratory studies were ordered per protocol. PetCO2 values were correlated with respiratory rate, PCO2, and pH. MEASUREMENTS AND MAIN RESULTS: One hundred twenty-one patients were monitored for 5.9 +/- 0.32 hrs. The average (mean +/- sd) initial values for pH were 7.08 +/- 0.18; respiratory rate, 35.1 +/ 9.1 breaths/min; PetCO2, 18.6 +/- 10.8 torr; and venous PCO2, 20.2 +/- 10.6 torr. At the conclusion of the observation period, averages were pH, 7.29 +/- 0.05; respiratory rate, 22.4 +/- 3.7 breaths/min; PetCO2, 35.3 +/- 5.8 torr; and venous PCO2, 36.8 +/- 5.3 torr. For all 592 observations, the correlations between PetCO2 and venous PCO2 (r =.92, p =.0001), PetCO2 and pH (r =.88, p =.0001), Petco2 and respiratory rate (r = -.79, p =.0001), and respiratory rate and pH (r = -.80, p =.0001) were statistically significant and the correlations with respiratory rate were inversely related to pH and PetCO2. The difference scores were not related to the average scores for initial readings (r = -.073, p =.43), final readings (r = -.124, p =.18), and overall readings (r =.057, p =.17). Limits of agreement between the two methods were established with PetCO2 lower than venous PCO2 with 95% limits of agreement 0.8 +/- 8.3 (2 sd) torr. CONCLUSIONS: PetCO2 monitoring of patients with diabetic ketoacidosis provides an accurate estimate of PCO2. Noninvasive PetCO2 sampling may be useful in patients with diabetic ketoacidosis to allow for continuous monitoring of patients. PMID- 14530765 TI - Pneumonia in the intensive care unit. AB - OBJECTIVE: To update the state-of-the-art on pneumonia in adult patients in the intensive care unit (ICU), with special emphasis on new developments in management. METHODS: We searched MEDLINE, using the following keywords: hospital acquired pneumonia, ventilator-associated pneumonia and healthcare-associated pneumonia, diagnosis, therapy, prevention. RESULTS: Interventions to prevent pneumonia in the ICU should combine multiple measures targeting the invasive devices, microorganisms, and protection of the patient. Once pneumonia develops, the appropriateness of the initial antibiotic regimen is a vital determinant of outcome. Three questions should be formulated: a) Is the patient at risk of methicillin-resistant Staphylococcus aureus?; b) Is Acinetobacter baumannii a problem in the institution?; and c) is the patient at risk of Pseudomonas aeruginosa? Antibiotic therapy should be started immediately and must circumvent pathogen-resistance mechanisms developed after previous antibiotic exposure. Therefore, antibiotic choice should be institution specific and patient oriented. Microbiologic investigation is useful on evaluating the quality of the respiratory sample and permits early modification of the regimen in light of the microbiologic findings. CONCLUSION: A decision tree outlining an approach to the evaluation and management of ventilator-associated pneumonia is provided. PMID- 14530766 TI - Managing hemorrhagic shock: fluids on the way out--drugs on the way in? PMID- 14530767 TI - Medical errors in the intensive care unit: can we find the black box before the patient crashes? PMID- 14530768 TI - Glutamine, a life-saving nutrient, but why? PMID- 14530769 TI - Xenon in intensive care. PMID- 14530770 TI - Clinical relevance of Acinetobacter baumannii ventilator-associated pneumonia. PMID- 14530771 TI - Hypertonic/hyperoncotic treatment for brain damage. PMID- 14530772 TI - Decompressive craniectomy. PMID- 14530773 TI - Noninvasive monitoring of the response to therapy during diabetic ketoacidosis: is end-tidal CO2 useful? PMID- 14530774 TI - Drotrecogin alfa (activated) administration: too many subgroups. PMID- 14530775 TI - Insulin dose or glycemic control for the critically ill? PMID- 14530776 TI - Evidence-based medicine. PMID- 14530777 TI - Open versus closed units: chaos versus a well-oiled machine. PMID- 14530778 TI - Primary (AL) hepatic amyloidosis: clinical features and natural history in 98 patients. AB - The liver is a common site of amyloid deposition in primary systemic amyloidosis. We reviewed the clinical features and natural history of patients with primary systemic amyloidosis and biopsy-proven hepatic involvement who were evaluated at Mayo Clinic from January 1, 1975, to December 31, 1997. The median age of the study group (68 men; 30 women) was 58.5 years. Seventy-one patients (72%) had involuntary weight loss. Hepatomegaly was found in 79 patients (81%). Eighty-two patients (89%) had proteinuria, and 81 patients (86%) had elevated serum alkaline phosphatase levels. Seventy-six patients (83%) had either a serum or urine monoclonal protein. Before liver biopsy, clinicians considered amyloidosis in the differential diagnosis for only 14 patients (26%). None of our patients experienced hepatic rupture or death due to liver biopsy, and only 4 (4%) bled after liver biopsy. The median survival of the 98 patients was 8.5 months. Predictors of a poor prognosis were congestive heart failure, elevated concentrations of bilirubin, and a platelet count greater than 500 x 109/L. In conclusion, clinicians should consider the diagnosis of primary hepatic amyloidosis in patients who present with involuntary weight loss or hepatomegaly. Other clues to the diagnosis include an unexplained elevated serum alkaline phosphatase level, proteinuria, and evidence for hyposplenism (for example, Howell-Jolly bodies on peripheral blood smear). Liver biopsy was safe. Some patients benefit from systemic chemotherapy. PMID- 14530779 TI - Morbidity and mortality in systemic lupus erythematosus during a 10-year period: a comparison of early and late manifestations in a cohort of 1,000 patients. AB - In the present study, we assessed the frequency and characteristics of the main causes of morbidity and mortality in systemic lupus erythematosus (SLE) during a 10-year period and compared the frequency of early manifestations with those that appeared later in the evolution of the disease. In 1990, we started a multicenter study of 1,000 patients from 7 European countries. All had medical histories documented and underwent medical interview and routine general physical examination when entered in the study, and all were followed prospectively by the same physicians during the ensuing 10 years (1990-2000).A total of 481 (48.1%) patients presented 1 or more episodes of arthritis at any time during the 10 years, 311 (31.1%) patients had malar rash, 279 (27.9%) active nephropathy, 194 (19.4%) neurologic involvement, 166 (16.6%) fever, 163 (16.3%) Raynaud phenomenon, 160 (16.0%) serositis (pleuritis and/or pericarditis), 134 (13.4%) thrombocytopenia, and 92 (9.2%) thrombosis. When the prevalences of the clinical manifestations during the initial 5 years of follow-up (1990-1995) were compared with those during the ensuing 5 years (1995-2000), most manifestations were found to be more frequent during the initial 5 years. Of the 1,000 patients, 360 (36%) presented infections, 169 (16.9%) hypertension, 121 (12.1%) osteoporosis, and 81 (8.1%) cytopenia due to immunosuppressive agents. Twenty-three (2.3%) patients developed malignancies; the most frequent primary localizations were the uterus and the breast.Sixty-eight (6.8%) patients died, and the most frequent causes of death were similarly divided between active SLE (26.5%), thromboses (26.5%), and infections (25%). A survival probability of 92% at 10 years was found. A lower survival probability was detected in those patients who presented at the beginning of the study with nephropathy (88% versus 94% in patients without nephropathy, p = 0.045). When the causes of death during the initial 5 years of follow-up (1990-1995) were compared with those during the ensuing 5 years (1995 2000), active SLE and infections (28.9% each) appeared to be the most common causes during the initial 5 years, while thromboses (26.1%) became the most common cause of death during the last 5 years.In conclusion, most of the SLE inflammatory manifestations appear to be less common after a long-term evolution of the disease, probably reflecting the effect of therapy as well as the progressive remission of the disease in many patients. Meanwhile, a more prominent role of thrombotic events is becoming evident, affecting both morbidity and mortality in SLE. PMID- 14530780 TI - Candidemia in a tertiary care cancer center: in vitro susceptibility and its association with outcome of initial antifungal therapy. AB - Since the 1990s, changing trends have been documented in species distribution and susceptibility to bloodstream infections caused by Candida species in cancer patients. However, few data are available regarding the association between in vitro antifungal susceptibility and outcome of candidemia in this patient population. We therefore evaluated the association of in vitro antifungal susceptibility and other risk factors with failure of initial antifungal therapy in cancer patients with candidemia. Candidemia cases in cancer patients from 1998 to 2001 (n = 144) were analyzed retrospectively along with their in vitro susceptibility to amphotericin B, fluconazole, and itraconazole (National Committee for Clinical and Laboratory Standards M27-A method). Patients were evaluable for outcome analysis if they received continuous unchanged therapy with either fluconazole or amphotericin B for >/=5 days. We excluded cases of mixed candidemia. In vitro susceptibility testing data of the first Candida bloodstream isolate were analyzed. Appropriate therapy was defined as that using an active in vitro antifungal for >/=5 days. For fluconazole susceptible-dose dependent Candida species, we defined appropriate therapy as a fluconazole dose of >/=600 mg/day. The Candida species distribution was 30% Candida albicans, 24% Candida glabrata, 23% Candida parapsilosis, 10% Candida krusei, 9% Candida tropicalis, and 3% other. Overall, amphotericin B was the most active agent in vitro, with only 3% of the isolates exhibiting resistance to it (>1 mg/L). Dose-dependent susceptibility to fluconazole and itraconazole was seen in 13% and 21% of the isolates, respectively, while resistance to fluconazole and itraconazole was seen in 13% and 26%, respectively.Eighty patients were evaluable for outcome analysis. In multivariate analysis, the following factors emerged as independent predictors of failure of initial antifungal therapy: leukemia (p = 0.01), bone marrow transplantation (p = 0.006), and intensive care unit stay at onset of infection (p = 0.02). Inappropriate antifungal therapy, as defined by daily dose and in vitro susceptibility, was not shown consistently to be a significant factor (it was significant in multivariate analysis, p = 0.04, but not in univariate analysis), indicating the complexity of the variables that influence the response to antifungal treatment in cancer patients with candidemia. PMID- 14530781 TI - A prospective multicenter study of Staphylococcus aureus bacteremia: incidence of endocarditis, risk factors for mortality, and clinical impact of methicillin resistance. AB - Our objectives were to determine the incidence of endocarditis in patients whose Staphylococcus aureus bacteremia was community-acquired, related to hemodialysis, or hospital-acquired; to assess clinical factors that would reliably distinguished between S. aureus bacteremia and S. aureus endocarditis; to assess the emergence of methicillin-resistant S. aureus (MRSA) as a cause of endocarditis; and to examine risk factors for mortality in patients with S. aureus endocarditis. We conducted a prospective observational study in 6 university teaching hospitals; we evaluated 505 consecutive patients with Staphylococcus aureus bacteremia. Thirteen percent of patients with S. aureus bacteremia were found to have endocarditis, including 21% with community-acquired S. aureus bacteremia, 5% with hospital-acquired bacteremia, and 12% on hemodialysis. Infection was due to MRSA in 31%. Factors predictive of endocarditis included underlying valvular heart disease, history of prior endocarditis, intravenous drug use, community acquisition of bacteremia, and an unrecognized source. Twelve patients with bacteremia had a prosthetic valve; 17% developed endocarditis. Unexpectedly, nonwhite race proved to be an independent risk factor for endocarditis by both univariate and multivariate analyses. Persistent bacteremia (positive blood cultures at day 3 of appropriate therapy) was identified as an independent risk factor for both endocarditis and mortality, a unique observation not reported in other prospective studies of S. aureus bacteremia. Patients with endocarditis due to MRSA were significantly more likely to have complicating renal insufficiency and to experience persistent bacteremia than those with endocarditis due to MSSA. The 30-day mortality was 31% among patients with endocarditis compared to 21% in patients who had bacteremia without endocarditis (p = 0.055). Risk factors for death due to endocarditis included severity of illness at onset of bacteremia (as measured by Apache III and Pitt bacteremia score), MRSA infection, and presence of atrioventricular block on electrocardiogram. Patients with S. aureus bacteremia who have community acquisition of infection, underlying valvular heart disease, intravenous drug use, unknown portal of entry, history of prior endocarditis, and possibly, nonwhite race should undergo echocardiography to screen for the presence of endocarditis. We recommend that blood cultures be repeated 3 days following initiation of antistaphylococcal antibiotic therapy in all patients with S. aureus bacteremia. Positive blood cultures at 3 days may prove to be a useful marker in promoting more aggressive management, including more potent antibiotic therapy and surgical resection of the valve in endocarditis cases. MRSA as the infecting organism should be added to the list of risk factors for consideration of valvular resection in cases of endocarditis. PMID- 14530782 TI - Staphylococcus aureus bacteremia: recurrence and the impact of antibiotic treatment in a prospective multicenter study. AB - Staphylococcus aureus bacteremia is associated with substantial morbidity. Recurrence is common, but incidence and risk factors for recurrence are uncertain. The emergence of methicillin resistance and the ease of administering vancomycin, especially in patients who have renal insufficiency, have led to reliance on this drug with the assumption that it is as effective as beta-lactam antibiotics, an assumption that remains open to debate. We initiated a multicenter, prospective observational study in 6 university hospitals and enrolled 505 consecutive patients with S. aureus bacteremia. All patients were monitored for 6 months and patients with endocarditis were followed for 3 years. Recurrence was defined as return of S. aureus bacteremia after documentation of negative blood cultures and/or clinical improvement after completing a course of antistaphylococcal antibiotic therapy. All blood isolates taken from patients with recurrent bacteremia underwent pulsed-field gel electrophoresis testing. Recurrence was subclassified as reinfection (different pulsed-field gel electrophoresis patterns) or relapse (same pulsed-field gel electrophoresis pattern).Forty-two patients experienced 56 episodes of recurrence (79% were relapses and 21% were reinfection). Relapse occurred earlier than reinfection (median, 36 versus 99 d, p < 0.06). Risk factors for relapse of S. aureus bacteremia included valvular heart disease, cirrhosis of the liver, and deep seated infection (including endocarditis). Nafcillin was superior to vancomycin in preventing bacteriologic failure (persistent bacteremia or relapse) for methicillin-susceptible S. aureus (MSSA) bacteremia. Failure to remove infected intravascular devices/catheters and vancomycin therapy were common factors in patients experiencing multiple (greater than 2) relapses. However, by multivariate analysis, only endocarditis and therapy with vancomycin (versus nafcillin) were significantly associated with relapse. Recurrences occurred in 9.4% of S. aureus bacteremias following antistaphylococcal therapy, and most were relapses. Duration of antistaphylococcal therapy was not associated with relapse, but type of antibiotic therapy was. Nafcillin was superior to vancomycin in efficacy in patients with MSSA bacteremia. PMID- 14530783 TI - Septic arthritis of the pubic symphysis: review of 100 cases. AB - We report a novel case of septic arthritis of the symphysis pubis due to Streptococcus pneumoniae and review 99 previously reported cases of infection of this joint. Typical features of pubic symphysis infection included fever (74%), pubic pain (68%), painful or waddling gait (59%), pain with hip motion (45%), and groin pain (41%). Risk factors included female incontinence surgery (24%); sports, especially soccer (19%); pelvic malignancy (17%); and intravenous drug use (15%). Septic arthritis of the pubic symphysis is often misdiagnosed as osteitis pubis, a sterile inflammatory condition. Causative organisms differed according to risk factors. Staphylococcus aureus was the major cause among athletes, Pseudomonas aeruginosa among intravenous drug users, and infections among patients with pelvic malignancies were usually polymicrobial, involving fecal flora. Patients with recent urinary incontinence surgery usually had monomicrobial infection, with no predominant pathogen. Since osteomyelitis is present in 97% of patients, we recommend antibiotic courses of 6 weeks' duration. Surgical debridement is required in 55% of patients. PMID- 14530784 TI - Enterococcal meningitis: a clinical study of 39 cases and review of the literature. AB - To describe the clinical features and outcome of enterococcal meningitis, we retrospectively reviewed the charts of 39 cases seen at 2 tertiary hospitals during a 25 years and collected 101 additional, previously reported cases for review. Among these 140 cases, there were 82 cases (59%) of postoperative meningitis and 58 cases (41%) of spontaneous meningitis. Eighty-six patients (61%) were adults and 54 (39%) were children. Patients with spontaneous meningitis had a higher frequency of community-acquired infection (50% versus 18%; p < 0.01), severe underlying diseases (67% versus 22%; p < 0.01), and associated enterococcal infection (29% versus 8%; p < 0.01) than patients with postoperative meningitis. The clinical presentation was similar in both groups, but patients with spontaneous infection had a higher frequency of bacteremia (58% versus 12%; p < 0.01), and a lower frequency of mixed infection (9% versus 29%; p < 0.01). Spontaneous meningitis in children was associated with a significantly lower frequency of fever, altered mental status, headache, and meningeal signs (p < 0.01), probably explained by the high proportion of neonates in this age-group. Most infections were caused by Enterococcus faecalis, which accounted for 76% of the isolates identified at the species level. Fifteen of the 25 cases due to Enterococcus faecium were produced by vancomycin-resistant strains. Most patients were treated with ampicillin, penicillin, or vancomycin, with or without aminoglycosides, for a median period of 18 days (range, 1-85 d). Overall mortality was 21%. The mortality rate was higher in spontaneous than in postoperative meningitis (33% versus 12%; p < 0.01), but was similar in patients treated with beta-lactams (18%), glycopeptides (14%), or other antibiotics (25%), as well as in patients treated with monotherapy (16%) or combination therapy (22%). An adverse outcome correlated significantly with advanced age, the presence of severe underlying diseases, associated enterococcal infection, bacteremia, septic shock, and the absence of fever at presentation. Shunt removal was associated with a lower mortality. Multivariate analysis showed that the presence of severe underlying diseases was the only prognostic factor associated with mortality (odds ratio = 6.8, 95% confidence intervals = 2.7-17.5, p < 0.01). PMID- 14530785 TI - Habitual betel quid chewing as a risk factor for cirrhosis: a case-control study. AB - Betel quid chewing, part of traditional Taiwanese culture, is common in 10%-20% of the human population worldwide. In this case-control study we assessed the independent and interactive role of habitual betel quid chewing and chronic hepatitis B virus (HBV) and hepatitis C virus (HCV) infection on risk of cirrhosis. Subjects enrolled included 210 pairs of sex- and age-matched cirrhotic patients and healthy controls. Information on risk factors was obtained through serologic examination of hepatitis B surface antigen (HBsAg) and antibodies to hepatitis C virus (anti-HCV), and a standardized personal interview with a structured questionnaire. Univariate analysis indicated that betel quid chewing, HBsAg+, anti-HCV+, alcohol drinking, and smoking are significant risk factors for cirrhosis. Multivariate analysis indicated that betel quid chewing (odds ratio [OR], 3.56), HBsAg (OR 20.37), and anti-HCV (OR 31.43) are independent risk factors for cirrhosis. Most betel quid chewers habitually drink alcohol. Although our analysis indicates that betel quid chewing acts independently from alcohol as a risk factor for cirrhosis, the confounding effect of alcohol cannot be excluded entirely by our study. There was an additive effect of the interaction between betel quid chewing and the presence of either HBsAg or anti-HCV. Moreover, a higher risk of cirrhosis was associated with longer duration of betel quid chewing and greater amount of betel quid consumed (each p for trend <0.0001). In conclusion, betel quid chewing appears to be an independent risk factor for cirrhosis. There is an additive interaction between betel quid chewing and chronic HBV/HCV infection. PMID- 14530787 TI - Introduction to anti-allergic properties of antihistamines. PMID- 14530788 TI - H1-Antihistamines: more relevant than ever in the treatment of allergic disorders. AB - Histamine is an important chemical mediator of inflammation, vasodilation, increased vascular permeability, decreased peripheral resistance, airway smooth muscle contraction, and sensory nerve stimulation causing itching. It also plays a significant role in neurotransmission and in cardiac function. In allergic rhinoconjunctivitis and urticaria, there is strong evidence for the role of H(1) antihistamine treatment. In asthma, additional dose-response studies, including higher doses of antihistamines than those used in allergic rhinitis, are needed to determine the role of antihistamines. In atopic dermatitis, the itch-relieving topical glucocorticoid-sparing effects of H(1)-antihistamines also require further documentation. The potential benefits of each H(1)-antihistamine should be weighed against the potential risks, and second-generation H(1)-antihistamines with excellent, well-documented safety records should be used in preference to older, less safe H(1)-antihistamines. Second-generation H(1)-antihistamines are more relevant than ever in the treatment of allergic disorders. PMID- 14530789 TI - Histamine: A mediator of inflammation. AB - Histamine and its receptors, including the recently discovered receptors (H(3) and H(4)), novel sources of histamine, and the place of histamine in mediator networks continue to be areas of great interest. The 4 major subtypes of histamine receptors, H(1) to H(4), differ in their location, second messengers, and histamine-binding characteristics. In addition, it would appear that different histamine receptor agonists and antagonists bind to different portions of the receptor complex. A fifth receptor subtype, the intracellular H(IC), has only been defined by its location within cell types that are not traditionally associated with histamine. In airway tissue, most cells express at least 1 subtype of histamine receptor; however, blockade of these receptors does not completely abolish the inflammatory response. In addition, some H(1) antihistamines might also exert anti-inflammatory effects by pathways independent of H(1)-receptor binding. Studies of selected second-generation H(1) antihistamines have shown that these agents inhibit the release of certain cytokines from basophils, acting at a point downstream from the calcium signaling pathway. It has not yet been determined whether this action represents a class effect. PMID- 14530790 TI - H1-receptors: localization and role in airway physiology and in immune functions. AB - Histamine H(1)-receptors are involved in the pathologic processes of allergy. Clinical trials of H(1)-receptor antagonists have demonstrated the efficacy of these agents in reducing the sneezing, pruritus, and rhinorrhea associated with allergic rhinitis. In the lung, H(1)-receptors mediate the bronchoconstrictive effects of histamine and increase vascular permeability, which lead to plasma exudation. H(1)-receptors are present on T cells, B cells, monocytes, and lymphocytes, and stimulation of these receptors induces pro-inflammatory effects. It has been suggested that a signal from the H(1)-receptor contributes to the antigen receptor-mediated signaling pathways that induce proliferative responses and lead to the production of cytokines and antibodies by T cells and B cells, respectively. It would appear, therefore, that the H(1)-receptor has a wider role in inflammatory processes than simply mediating the actions of histamine. PMID- 14530791 TI - Efficacy and safety profile of fexofenadine HCl: a unique therapeutic option in H1-receptor antagonist treatment. AB - Results of head-to-head comparative trials suggest that fexo-fenadine might offer distinct advantages compared with other antihistamines. Fexofenadine is highly selective for peripheral H(1)-receptors and does not cross the blood-brain barrier, as shown by positron emission tomography. These data support findings that fexofenadine is nonsedating and does not impair performance or driving ability, even at very high doses. In addition, fexofenadine does not interact with muscarinic receptors, which might offer a potential advantage compared with desloratadine, the recently approved active metabolite of loratadine. Fexofenadine is devoid of adverse cardiac effects, and changes in electrocardiogram parameters are not significantly different from those observed with placebo. Fexofenadine has also been shown to have a favorable effect on nasal congestion. This therapeutic advantage might be related to its significant antiallergic properties, ie, the demonstrated ability of fexofenadine to blunt the inflammatory effects of preformed and de novo synthesized mediators at clinically relevant doses in vivo and in vitro. Cumulatively, these benefits distinguish fexofenadine from other antihistamines and make it an optimum therapeutic option for treating allergy-mediated respiratory and dermatologic diseases. PMID- 14530793 TI - The histamine-cytokine network in allergic inflammation. AB - Histamine is synthesized and released by human basophils, mast cells, and neurons. Its pleiotropic effects are mediated by the activation of 4 receptors: H(1), H(2), H(3), and H(4). With the advent of selective antagonists (the antihistamines widely used to treat allergic disorders), the H(1)-receptor was the first member of the receptor family to be pharmacologically defined. Increasing evidence indicates that, in addition to exerting immediate vascular and bronchial responses, histamine might modulate the immune reaction by interacting with T cells, macrophages, basophils, eosinophils, and monocytes. We have shown that, in vitro, histamine induces a concentration-dependent release of IL-6 and beta-glucuronidase from macrophages isolated from the human lung parenchyma. These effects are inhibited by fexofenadine, an H(1)-receptor antagonist, but not by ranitidine, an H(2)-receptor antagonist. This observation raises the possibility that long-term treatment with fexofenadine might have beneficial effects on immune dysregulation and tissue damage/remodeling associated with histamine-mediated macrophage activation. PMID- 14530792 TI - Effects of fexofenadine and other antihistamines on components of the allergic response: adhesion molecules. AB - Intercellular adhesion molecules (ICAMs), in particular ICAM-1, appear to play a crucial role in the recruitment and migration of inflammatory cells to the site of an allergic reaction. Glucocorticoids and allergen-specific immunotherapy have been shown to exert effects on selected components of this system, both in vitro and in vivo, but further research is required to better understand the effects of these therapies. Nasal and conjunctival challenge models (including natural and experimental allergen exposure) represent useful and safe tools for studying the activity of antiallergy drugs in vivo. These tests allow the investigation of a wide variety of parameters including inflammatory infiltrate, ICAM-1 expression, and changes in the concentration of soluble inflammatory mediators. With these tools, anti-inflammatory activity related to the modulation of epithelial cell adhesion molecules has been demonstrated in vivo for several H(1)-receptor antagonists (azelastine, cetirizine, loratadine, levocabastine, oxatomide, and terfenadine). Fexofenadine is a nonsedating, long-acting antihistamine with highly selective H(1)-receptor antagonist activity and a particularly favorable safety profile. In addition, fexofenadine has proven anti-inflammatory activity and has been shown to inhibit a number of mediators at clinically relevant concentrations, including in vitro inhibition of ICAM-1 expression on conjunctival and nasal epithelial cells. PMID- 14530795 TI - Prospects for antihistamines in the treatment of asthma. AB - Antihistamines have been evaluated as potential therapies for asthma for more than 50 years. With first-generation compounds, side effects prevented effective dosing. By reviewing published studies of the effectiveness of terfenadine, cetirizine, and loratadine in clinical asthma, evidence for the relevant effect of the second-generation antihistamines on bronchial asthma can be found. Terfenadine, at doses of 120 or 180 mg twice a day, reduced symptoms and improved pulmonary function in mild and moderate allergic asthma but was ineffective in severe perennial asthma. Fexofenadine at doses used for allergic rhinitis had little effect on seasonal allergic asthma. Research is ongoing to determine the effects of higher doses of fexofenadine. In 5 studies, cetirizine at doses of 10 to 20 mg, once or twice daily, consistently improved asthma symptoms compared with placebo or terfenadine 60 mg twice a day in 2 cases, whereas in 2 studies, loratadine at doses of 10 to 20 mg daily has not produced significant improvement in asthma. However, loratadine 5 mg combined with 60 mg of pseudo-ephedrine twice a day significantly improved both asthma symptoms and peak expiratory flow. Similarly, the combination of loratadine 20 mg and the leukotriene-receptor antagonist montelukast improved asthma symptoms, peak expiratory flow, and beta agonist use over montelukast alone. Therefore, there might be a role for second- and third-generation antihistamines in treating mild and moderate asthma, which might require administering doses greater than those commonly used to treat allergic rhinitis. If higher doses are sedative, the addition of decongestants or leukotriene-receptor antagonists might enhance the effects of lower doses of the antihistamines. PMID- 14530794 TI - Effects of fexofenadine on T-cell function in a murine model of allergen-induced airway inflammation and hyperresponsiveness. AB - There is renewed interest in antihistamines for the treatment of allergic asthma. A growing body of literature has shown that the newer compounds can affect inflammatory cell accumulation and cytokine/chemokine production. In a murine model of allergen-induced airway inflammation and hyperresponsiveness, the ability of fexofenadine to affect these outcomes was tested in a primary sensitization and challenge model and after treatment of donor mice before the adoptive transfer of T cells into recipients receiving limited allergen exposure. Mice were sensitized and challenged with allergen (ovalbumin). Airway function after inhaled methacholine was monitored in parallel to the assessment of tissue and airway inflammation and cytokine production. In further experiments, lung T lymphocytes from sensitized/challenged donor mice were transferred into naive recipients before limited airway challenge with the allergen. Administration of fexofenadine before challenge but after sensitization was effective in preventing tissue eosinophilia and airway hyperresponsiveness. Moreover, the treatment of donor mice with fexofenadine before transfer of lung T cells effectively prevented airway hyperresponsiveness and eosinophilia in naive mice exposed to limited airway challenge. These data therefore support the potential for fexofenadine in the treatment of allergen-induced airway hyperresponsiveness and inflammation. PMID- 14530796 TI - Antinociception and the new COX inhibitors: research approaches and clinical perspectives. AB - New generations of cyclooxygenase (COX) inhibitors are more potent and efficacious than their traditional parent compounds. They are also safer than the classic non-steroidal anti-inflammatory drugs (NSAIDs) and are starting to be used not only for low to moderate intensity pain, but also for high intensity pain. Three different strategies have been followed to improve the pharmacological profile of COX inhibitors: 1. Development of COX-2 selective inhibitors. This is based on the initial hypothesis that considered COX-2 as the enzyme responsible for the generation of prostaglandins only in inflammation, and, therefore, uniquely responsible for inflammation, pain and fever. Initial expectations gave rise to controversial results, still under discussion. The second generation of these compounds is being developed and should contribute to clarifying both their efficacy and the specific functions of the COX enzymes. 2. Modified non-selective COX inhibitors. Molecules like nitro-NSAIDs or tromethamine salt derivatives have been synthesized considering that both COX-1 and COX-2 are responsible for the synthesis of prostaglandins involved either in homeostatic functions or inflammation. Nitroaspirin, nitroparacetamol or dexketoprofen trometamol are some examples of molecules that are already showing an important clinical efficacy. The modifications performed in their structures seem to lower the unwanted side effects as well as to enhance their analgesic efficacy. 3. Combined therapy of classic NSAIDs with other drugs. This strategy looks for improvements in the incidence of adverse effects or to take advantage of the synergistic enhancement of their therapeutic effects. Some of the molecules resulting from these strategies are very valuable as therapeutic agents and open a wide range of possibilities in the treatment of high intensity pain, including neuropathic pain, and opiate sparing therapy. PMID- 14530797 TI - Development of the nitrone-based spin trap agent NXY-059 to treat acute ischemic stroke. AB - The only current FDA-approved treatment for acute ischemic stroke is thrombolysis with tissue plasminogen activator (tPA). However, there are numerous shortcomings to tPA treatment including an increased incidence of intracerebral hemorrhage (ICH) and a short therapeutic window (3-6 h). In recent years, studies have attempted to identify new therapeutics that might be neuroprotective following ischemic strokes. Free radical scavenging spin trap agents have been proposed as potential candidates for stroke therapy because of the hypothesized role of free radicals in the progression of stroke and ischemia-induced neurodegeneration. Novel spin trap agents like (disodium-[(tert-butylimino) methyl] benzene-1,3 disulfonate N-oxide (NXY-059) are of particular interest, not only because they are broad-spectrum nitrone-based free radical scavengers, but also because of their safety profile in humans. Moreover, the rationale for developing NXY-059 for the treatment of acute ischemic stroke is further supported by the drug's reported neuroprotective effects. In addition, NXY-059 may represent a useful adjunct stroke therapy to tPA, since preclinical studies have demonstrated that NXY-059 increases the therapeutic window for tPA and lowers the occurrence of tPA induced ICH. PMID- 14530798 TI - Trialkylglycines: a new family of compounds with in vivo neuroprotective activity. AB - Glutamate neurotoxicity is involved in the pathogenesis of neurodegenerative disorders such as Huntington's, Parkinson's and Alzheimer's diseases. It plays also a major role in the neuronal damage that occurs in brain ischemia and head trauma. Finding molecules that prevent or reverse glutamate neurotoxicity (excitotoxicity) is, therefore, of great interest. Strategies aimed at this end include the screening of libraries of compounds synthesized by combinatorial chemistry to find molecules that prevent neuronal death in vitro and in vivo. A library of trialkylglycines was screened to assess whether they prevent glutamate induced neuronal death in primary cultures of cerebellar neurons. Two types of trialkylglycines have been found that significantly reduce the incidence of glutamate-induced neuronal death. The first type includes two compounds (referred to as 6-1-2 and 6-1-10) that efficiently prevent glutamate or NMDA-induced neuronal death. They also prevent excitotoxicity in vivo as assessed by using two animal models of excitotoxicity: acute intoxication with ammonia and a model of cerebral ischemia in rats. Trialkylglycines 6-1-2 and 6-1-10 prevent ammonia induced (NMDA receptor-mediated) death of mice and neuronal degeneration in the model of cerebral ischemia. The trialkylglycines of the second type act as open channel blockers of the NMDA receptor. The first group of trialkylglycines does not block NMDA receptor channels and does not affect the glutamate-nitric oxide cGMP pathway. Their molecular target has not yet been identified. These two types of trialkylglycines (especially those that do not affect NMDA receptor function) might represent effective drugs for the treatment of neurodegeneration. They are likely to be well tolerated and have fewer side effects than NMDA receptor antagonists. PMID- 14530799 TI - The neuropharmacological basis for the use of memantine in the treatment of Alzheimer's disease. AB - Memantine has been demonstrated to be safe and effective in the symptomatic treatment of Alzheimer's disease (AD). While the neurobiological basis for the therapeutic activity of memantine is not fully understood, the drug is not a cholinesterase inhibitor and, therefore, acts differently from current AD therapies. Memantine can interact with a variety of ligand-gated ion channels. However, NMDA receptors appear to be a key target of memantine at therapeutic concentrations. Memantine is an uncompetitive (channel blocking) NMDA receptor antagonist. Like other NMDA receptor antagonists, memantine at high concentrations can inhibit mechanisms of synaptic plasticity that are believed to underlie learning and memory. However, at lower, clinically relevant concentrations memantine can under some circumstances promote synaptic plasticity and preserve or enhance memory in animal models of AD. In addition, memantine can protect against the excitotoxic destruction of cholinergic neurons. Blockade of NMDA receptors by memantine could theoretically confer disease-modifying activity in AD by inhibiting the "weak" NMDA receptor-dependent excitotoxicity that has been hypothesized to play a role in the progressive neuronal loss that underlies the evolving dementia. Moreover, recent in vitro studies suggest that memantine abrogates beta-amyloid (Abeta) toxicity and possibly inhibits Abeta production. Considerable attention has focused on the investigation of theories to explain the better tolerability of memantine over other NMDA receptor antagonists, particularly those that act by a similar channel blocking mechanism such as dissociative anesthetic-like agents (phencyclidine, ketamine, MK-801). A variety of channel-level factors could be relevant, including fast channel-blocking kinetics and strong voltage-dependence (allowing rapid relief of block during synaptic activity), as well as reduced trapping (permitting egress from closed channels). These factors may allow memantine to block channel activity induced by low, tonic levels of glutamate--an action that might contribute to symptomatic improvement and could theoretically protect against weak excitotoxicity--while sparing synaptic responses required for normal behavioral functioning, cognition and memory. PMID- 14530803 TI - Continuous NSW health survey: quarterly report on health status, health behaviours, and risk factors. PMID- 14530801 TI - The Australian study of health and relationships: results for central Sydney, inner-eastern Sydney, and New South Wales. PMID- 14530804 TI - New South Wales Health Aboriginal Health Impact Statement: references and resources about Aboriginal people and Aboriginal health. PMID- 14530805 TI - Addressing disadvantage and health: the Mount Druitt pilot study. PMID- 14530806 TI - FactSheet: meningococcal disease. PMID- 14530808 TI - Epstein-Barr virus expression in Hodgkin's lymphoma in Kuwait. AB - The epidemiology of Hodgkin's lymphoma (HL) shows wide geographic variation in histological subtypes and in its association with the Epstein-Barr virus (EBV). The proportion of EBV positive HL is low in industrialized countries, high in non industrialized countries and intermediate in early-industrialized countries. Reports from the Persian Gulf and Middle East are very limited. The aim of this study was to determine the epidemiology of HL in Kuwait, an early-industrialized country in the Persian Gulf, and to delineate the extent of its association with EBV. We reviewed 134 cases of HL for histological classification and demographic data. 107 cases were examined for the presence of EBV using immunohistochemistry (IHC) for the latent membrane protein I (LMPI) and in-situ hybridization (ISH) for EBVencoded RNA (EBER). 70.4% of the patients were males and 29.6% were females. The male: female ratio was 2.4:1. The mean age was 30.6 years (range, 4 71 years). Mixed cellularity HL (MCHL) was the most common subtype (45.5%), followed by nodular sclerosis (37.3%), nodular lymphocyte predominant (6.7%), lymphocyte rich (3%) and lymphocyte depletion (3%). 4.5% of cases were unclassifiable. EBV expression was seen in 56%, was significantly higher in MCHL, in children, and in males. Our findings suggest that the frequency of EBV expression in HL in Kuwait is similar to other early-industrialized countries. Further research from other countries in the Persian Gulf and the Middle East should shed more light on the epidemiology of HL and its relation to EBV in this region. PMID- 14530807 TI - Brain-metastatic melanoma: a neurotrophic perspective. AB - The brain is a unique microenvironment enclosed by the skull and maintaining a highly regulated vascular transport barrier. To metastasize to the brain, malignant tumor cells must attach to microvessel endothelial cells, invade the blood-brain barrier (BBB), and respond to brain survival and growth factors. Neurotrophins (NT) are important in brain invasion because they stimulate this process. In brain-metastatic melanoma cells, NT can promote invasion by enhancing the production of extracellular matrixdegradative enzymes such as heparanase, an enzyme capable of locally destroying both the extracellular matrix and the basement membrane of the BBB. We have examined human and murine melanoma cell lines exhibiting varying abilities to form brain metastases, and have found that they express low-affinity neurotrophin receptor p75NTR in relation to their brain metastatic potentials. They do not, however, express trkA, the gene encoding the tyrosine kinase receptor TrkA, the high-affinity receptor for nerve growth factor (NGF), the prototypic NT. Presence of functional TrkC, the putative receptor for the invasion-promoting neurotrophin NT-3, was also expressed in these cells. Brain-metastatic melanoma cells can also produce autocrine factors and inhibitors that influence their growth, invasion, and survival in the brain. Synthesis of these factors may influence NT production by brain cells adjacent to the neoplastic invasion front, such as oligodendrocytes and astrocytes. In brain biopsies, we observed increased amounts of NGF and NT-3 in tumor-adjacent tissues at the invasion front of human melanoma tumors. Additionally, we found that astrocytes contribute to the brain-metastatic specificity of melanoma cells by producing NT-regulated heparanase. Trophic, autocrine, and paracrine growth factors may therefore determine whether metastatic cells can successfully invade, colonize, and grow in the central nervous system (CNS). PMID- 14530809 TI - Adjuvant therapy of breast cancer with docetaxel-containing combination (TAC). AB - The adjuvant chemotherapy of breast cancer changed in the past two decades. Docetaxel containing regimens are highly active in metastatic breast cancer. A logical approach was their incorporation into trials of early breast cancer adjuvant therapy. The authors present the Hungarian interim analysis and experience with the BCIRG 001 randomized, multicentric, phase III clinical trial comparing TAC (docetaxel, doxorubicin, cyclophosphamide) and FAC (5-fluorouracil, doxorubicin, cyclophosphamide) in the adjuvant treatment of node positive breast cancer patients. The results are presented compared to the international data. Three Hungarian centers - Szt. Margit Hospital, Budapest, National Institute of Oncology, Budapest, Petz Aladar Hospital, Gyor - participated in the international trial. Between June 1997 and June 1999, 61 patients with node positive breast cancer were enrolled in the study after the surgery. Thirty-four patients were randomized to TAC (75/50/500 mg/m2 6xq3wk) and 27 patients were randomized to FAC (500/50/500 mg/m2 6x q3wk) chemotherapy, with prospective stratification by node (1-3, 4+). Patients with hormone receptor positive tumors received tamoxifen for 5 years after the chemotherapy. Radiotherapy was performed after the 6th cycle of chemotherapy. 33 months of follow up was performed. In both arms the hematological toxicity was more frequent. The TAC group showed a higher incidence of neutropenia (76%) compared to the FAC (22%), as well as a higher incidence of febrile neutropenia (26 % versus none), without grade 3-4 infection and there was no cases of septic death. More grade 3-4 nausea and vomiting was observed in the FAC group. At three years follow up, results indicated improvement in disease-free survival (88% vs. 76%) in favour of TAC, and similar tendency was observed in the case of overall survival (97% vs. 88%). Based on the international data analysis TAC was superior to FAC chemotherapy, the results show statistically significant differences between the two arms. This benefit with TAC was seen regardless of hormone receptor status. Additional follow up data will evaluate the role of TAC in the adjuvant setting of early breast cancer treatment. PMID- 14530810 TI - Increased expression of vascular endothelial growth factor (VEGF) in bone marrow of patients with myeloproliferative disorders (MPD). AB - Angiogenesis is a multistep process of the development of capillaries from established blood vessels. Angiogenesis probably plays a significant role in the development and progression of hematopoietic malignancies. Higher microvascular density and increased serum levels of proangiogenic factors such as vascular endothelial growth factor (VEGF) or basic fibroblasts growth factor (bFGF) have been reported in acute and chronic leukemias, myeloproliferative and myelodysplastic disorders, multiple myeloma and lymphomas. The microvessel density of bone marrow stroma in myeloproliferative disorders is increased and VEGF is considered as the most potent endothelial cell activator. The purpose of this study was to examine the expression of VEGF in bone marrow of patients with MPD. 60 paraffinembedded bone marrow core biopsy specimens from newly diagnosed patients with MPD were evaluated. In addition 10 bone marrow core biopsy specimens from adult patients without evidence of malignancy were used as controls. Bone marrow sections were stained immunohistochemically for VEGF (PharMingen, USA). Obtained data show that MPD are associated with an increased expression of VEGF in the bone marrow. This observation support previous studies suggesting that angiogenesis may play a role in the pathophysiology of myeloproliferative disorders. Clinical significance of this phenomenon needs further investigation however thus provides rationale for use of angiogenesis inhibitors in MPD therapy. PMID- 14530812 TI - Influence of obesity on lymph node recovery from rectal resection specimens. AB - Careful lymph node dissection from colorectal resection specimens is important procedure for cancer staging. Present study intended to assess the impact of surgical technique and patient's obesity on this process. Number of lymph nodes harvested by manual dissection from resection specimens of 141 patients with rectal cancer and the rate of nodal metastases were analyzed and compared in different groups of patients selected by length of resection specimen and body mass index. The median and mean number of lymph nodes found per patient were 6 and 6.7. The shorter resection specimens (<16 cm after formalin fixation) yielded significantly lower number of nodes than those with length > 16 cm (5.7 versus 7.9). Most significant reduction in mean number of lymph nodes was observed in obese patients with short specimens (4.8). This subset of patients presented the lowest rate of nodal metastases (38%). The surgical technique seems to be an important factor for lymph node recovery from rectal resections specimens. The patient's obesity had an unfavourable impact on this procedure. Standardized surgery and histopathological examination are needed even in non-specialized centers to harvest adequate number of lymph nodes. PMID- 14530811 TI - Correlation between BCR-ABL expression and tumor burden is restricted to the transition from minor to major cytogenetic response in interferon treated CML patients. AB - The interferon treatment of chronic myeloid leukaemia has been monitored by investigating the tumour burden as revealed by fluorescence in situ hybridization and the expression of BCR-ABL chimera determined by quantitative reverse transcription polymerase chain reaction. These parameters were obtained from the peripheral blood of 51 untreated and 104 follow-up patient samples. Poor correlation (r=0.31) was found between BCR-ABL expression and tumor load in all samples as well as in untreated patients, and this correlation was even less in all follow-up cases (r=0.28). Regarding chimera expression five order of magnitude difference existed in the untreated patients and this value dropped to two in those with complete cytogenetic response. Only the major and the complete cytogenetic response groups differed significantly (p<0.001) in the BCR-ABL expression from that of patients at diagnosis. Among the different cytogenetic response groups the only significant difference (p<0.01) in the BCRABL expression was obtained between the major and the minor responders. In the individual patients not only correlated changes of residual tumour mass and chimera expression, but mainly independent changes of these two parameters were observed. This indicates that the BCR-ABL expression and the tumor burden are largely independent variables. PMID- 14530813 TI - Clinical significance of sentinel lymph node involvement in malignant melanoma. AB - In the period 1997-2002, sentinel lymph node (SLN) surgery was performed on 179 primary skin melanoma patients, one to two months after the removal of the primary. Staining with patent blue was combined with an isotope technique. Histological evaluation of the sentinel lymph nodes was performed in serial sections. Immunohistochemical detection of S100, HMB-45, or Melan-A was used in the case of suspected micrometastases. Demonstration of positive sentinel lymph node was followed, preferably within 2-3 weeks, by regional block dissection. In these cases interferon-a2 in low doses or BCG immune therapy were applied as adjuvant therapy. Bimonthly follow-up of the patients included physical examination and the use of imaging techniques as specified in the melanoma protocol. Sentinel lymph node surgery was successful in 177/179 cases (98%). Positive sentinel lymph node was identified in 26/177 patients (14.7%). In node positive patients the thickness of the primary tumour was significantly greater than that of node negative ones (p<0.00001). Patients with micrometastases had significantly poorer symptom-free and overall survival by the Mantel-Cox test than those of the other group (p=0.0001 and p=0.0007 respectively). Comparison of the tumor thickness and positive SLN by discriminance analysis, yielded 81.7% and 79.9%, respectively for correct classification rates. Based on our study and data from the literature, we suggest SLN-positivity as equally strong poor prognosis factor for skin melanoma as the tumor thickness. PMID- 14530814 TI - Colliding / concomitant tumors of the intestine: report of 3 cases. AB - Collision/concomitant tumors of the intestine involving lymphomas are very rare. For these cases molecular genetic analyses are valuable diagnostic adjuncts. We report one collision tumor of the rectum (adenocarcinoma and peripheral T-cell lymphoma, unspecified), and two cases of concomitant tumors (carcinoma in the cecum and lymphoma in the ileum; carcinoma in the sigmoid and lymphoma in the ileum). The collision tumor (poorly differentiated adenocarcinoma and a peripheral T-cell lymphoma, unspecified) showed a variable proportion of the anaplastic tumor cells expressing lymphatic markers as well as cytokeratin. Only polymerase chain reaction (PCR) analysis revealing T-cell monoclonality of the anaplastic part of the colliding tumor allowed the correct diagnosis. In the second case, a moderately differentiated adenocarcinoma in the cecum with a concomitant B-cell Non-Hodgkin lymphoma in the ileum, PCR analysis was non contributory. In the third case (adenocarcinoma in the sigmoid colon and a follicular center lymphoma in the ileum) PCR analysis revealed gene rearrangement of the immunoglobulin heavy chain gene. We would like to emphasize that collision and concomitant tumors of the gut are rare and that molecular genetic analysis may be mandatory for correct diagnosis. It is our impression, that these tumors may be diagnosed more often in the intestinal tract if molecular genetic analysis and immunohistochemistry are used routinely, at least for all anaplastic tumors. PMID- 14530816 TI - Vulvar syringoma aggravated by pregnancy. AB - Syringoma is a benign tumors of eccrine sweat gland. They appear as multiple, tiny, firm, skin-colored papules. Vulvar involvement of syringoma is rare. Only 24 cases with vulvar syringoma have been previously reported in the literature. The majority of patients with vulvar syringomas are asymptomatic. A case of syringoma of the vulva exacerbated during pregnancy is presented. The case appears remarkable for the experienced aggravated pruritic symptoms of the patient during her pregnancy. PMID- 14530815 TI - Renal medullary carcinoma in a six-year-old boy with sickle cell trait. AB - Renal medullary carcinoma (RMC), an aggressive malignant epithelial neoplasm, first emerged as a distinct clinicopathologic entity in 1995. It affects individuals 40 years of age or younger and is strongly associated with sickle cell disease or trait. The majority of patients with RMC have widely disseminated disease at the time of diagnosis and most fail to respond to both chemotherapy and radiotherapy. Mortality approaches 100%, and death usually occurs within a few months to a year of diagnosis. We report a 6-year-old African-American boy with a history of gross hematuria who died four weeks after diagnosis of disseminated metastatic disease. Autopsy showed a 4.4-cm renal mass with metastases to the contra lateral kidney, liver, lungs and multiregional lymph nodes. RMC should be included in the differential diagnosis of any patient 40 years old or younger with a history of hemoglobinopathy and gross hematuria and/or abdominal or flank pain. A brief discussion of the differential diagnosis, histogenesis and treatment is presented in this study. PMID- 14530817 TI - Endobronchial large cell neuroendocrine carcinoma. AB - Lung tumors with neuroendocrine morphology include typical carcinoid, atypical carcinoid, large cell neuroendocrine carcinoma, and small cell carcinoma. The World Health Organization emphasizes the importance of mitotic count in differentiating these tumors. We studied the case of a 58-year-old male nonsmoker with recurrent pneumonia and an endobronchial mass, which was removed by right middle lobectomy. The patient was alive with no recurrent disease at 36-month follow-up. Histologically, the tumor showed well developed neuroendocrine morphology but contained up to 20 mitoses per 10 high-power fields and was therefore diagnosed as a large cell neuroendocrine carcinoma. However, several features, including the carcinoid-like morphology and endobronchial location of the tumor, absence of smoking history, and promising clinical course, were more characteristic of an atypical carcinoid than of a large cell neuroendocrine carcinoma. It may be necessary to redefine histologic criteria to allow a higher mitotic rate for classification as an atypical carcinoid. PMID- 14530818 TI - Malignant peripheral nerve sheath tumors associated with neurofibromatosis type 1. AB - We report 4 cases of malignant peripheral nerve sheath tumors (MPNST) with neurofibromatosis type 1 (NF1). Mean age was 29.5. Two of them had a family history. Three of them were male. All of them had enlarging mass and pain in the background of neurofibromas. Locations were popliteal, thigh and forearm. The masses were greater than 5 cm in diameter in each case. In two cases the mass was showing continuity with a nerve. One patient had a nonossifying fibroma as well as a MPNST. Wide excision and radiotherapy were applied to three of the patients. One of them did not take any therapy after surgical resection. Two of the patients died of lung metastases after a mean period of 12.5 months. In a majority of NF1 patients MPNST emerges from a preexisting neurofibroma. The patients with NF1 are at greatest risk for developing sarcomas, so they should be followed closely. PMID- 14530819 TI - Respiratory acidosis sans acidemia. PMID- 14530820 TI - Contemplating guidelines. PMID- 14530822 TI - Re-evaluation of acid-base prediction rules in patients with chronic respiratory acidosis. AB - RATIONALE: The prediction rules for the evaluation of the acid-base status in patients with chronic respiratory acidosis, derived primarily from an experimental canine model, suggest that complete compensation should not occur. This appears to contradict frequent observations of normal or near-normal pH levels in patients with chronic hypercapnia. METHODS: Linear regression analysis was used to estimate the relationships between arterial pH, bicarbonate and partial pressure of carbon dioxide (PCO2) from 18 separate arterial blood gas measurements in 18 clinically stable outpatients with chronic hypercapnic respiratory failure from chronic obstructive lung disease, and without clinical conditions or medications likely to cause a primary metabolic alkalosis. RESULTS: The PCO2 ranged from 45 mmHg to 77 mmHg, and pH ranged from 7.37 to 7.44. In only three of the arterial blood gas measurements were the pH values lower than 7.38. From the regression equations derived from these measurements, the pH decreased by 0.014 for each 10 mmHg increase in the PCO2, and the bicarbonate level increased by 5.1 mmol/L. These values are quite different from a decrease in pH of 0.03 and an increase in bicarbonate of 3.5 mmol/L predicted using the rules derived from the canine model. CONCLUSIONS: In patients with chronic stable hypercapnia, acid-base compensatory mechanisms appear to be more effective than would be predicted using the classic rules. PMID- 14530824 TI - Economic issues in the use of office spirometry for lung health assessment. AB - The National Lung Health Education Program (United States) has recently recommended using office spirometry to screen for subclinical lung disease in adult smokers. No published studies evaluate the economic consequences of this recommendation. This review article outlines the issues that must be considered when evaluating the costs and health benefits of office spirometry. Much of the available data on the effectiveness of screening is from studies that included smoking cessation interventions, making it difficult to determine the effects of screening alone. The sensitivity and specificity of screening spirometry are not known, but may not be important in the economic model, because even false positive test results are beneficial if they lead to smoking cessation. Costs to be considered include those of spirometry itself, of implementing and maintaining screening and smoking cessation programs, and of their consequences, ie, reduced morbidity (lower short term health care costs) and mortality (perhaps higher long term health care costs). Despite these unique challenges, data are available to perform economic analyses regarding screening spirometry. Such analyses should play a role in future clinical policy making. Even modest quit rates attributable to screening spirometry may result in highly favourable cost effectiveness ratios. PMID- 14530825 TI - Pancoast's syndrome due to metastatic carcinoma from the stomach. AB - We describe here a case of Pancoast's syndrome due to metastatic carcinoma from the stomach. Although obtaining a tissue diagnosis is often difficult with apical lesions, transbronchial or percutaneous needle biopsy is the procedure of choice since a certain number of these cases are potentially curable. PMID- 14530826 TI - Recurrent embolotherapy in Dieulafoy's disease of the bronchus. AB - Dieulafoy's disease is a vascular anomaly characterized by the presence of a dysplastic artery that is related to an epithelial ulcer. The French surgeon Georges Dieulafoy first described it in 1898. Most frequently, it is a gastrointestinal condition, but occurrence in the bronchus has been reported in a few cases. The case of a 52-year-old man with massive hemoptysis, for which he underwent successful embolotherapy 10 years previously, is described. Over the next 10 years, he had several hospital admissions due to hemoptysis, and he underwent successful embolotherapy on each occasion. This case report underlines the importance of bronchial arteriography as the investigation of choice for massive hemoptysis. PMID- 14530828 TI - Left-handedness as a risk factor for fractures. AB - Left-handedness has been associated with increased fracture risk in a small number of previous studies. This study reports risks for fractures at the proximal humerus, distal forearm, pelvis, foot, and shaft of the tibia/fibula according to handedness in a case-control study conducted from October 1996 to May 2001 among members of Northern California Kaiser Permanente. Handedness was assessed by questionnaire for 2,841 cases and 2,192 controls, and subjects were categorized as left-handed, right-handed, ambidextrous, or forced to switch from the left to the right hand. Compared to right-handedness, left-handedness was most strongly associated with an increased risk for proximal humerus fractures (adjusted odds ratio (OR)=2.00, 95% confidence interval (CI) 1.33 to 3.01) and less definitively with fractures of the distal forearm (adjusted OR=1.28, 95% CI 0.92 to 1.80), foot (adjusted OR=1.17, 95% CI 0.82 to 1.65), and pelvis (adjusted OR=1.40, 95% CI 0.71 to 2.74). Ambidextrous individuals had elevated risks for fractures of the distal forearm (adjusted OR=2.99, 95% CI 1.42 to 6.30), foot (adjusted OR=2.59, 95% CI 1.13 to 5.97), shaft of the tibia/fibula (adjusted OR=3.91, 95% CI 1.01 to 15.17), and proximal humerus (adjusted OR=2.37, 95% CI 0.85 to 6.65) when compared with right-handed individuals. Those individuals forced to use the right hand demonstrated no increased risk for fractures at any site. These results suggest that handedness does influence fracture risk, but the reasons for this increased risk are unclear. PMID- 14530830 TI - Pelvic anatomy and the surgeon. PMID- 14530829 TI - Definition of a population-specific DXA reference standard in Italian women: the Densitometric Italian Normative Study (DINS). AB - Osteoporosis is currently defined on the basis of the T-score by dual-energy X ray absorptiometry (DXA). Despite its limitations, this definition is applied worldwide. However, the normal values provided by manufacturers may not be fully representative of specific local populations. So far, there are no normative data in the Italian population using Hologic densitometers. The Densitometric Italian Normative Study (DINS) is an ongoing multi-center study that aims to establish reference values for bone densitometry with dual-energy X-ray absorptiometry (DXA) in the male and female Italian population. In this paper we report the results of the lumbar vertebrae (L2-L4) and proximal femur in 1,622 women aged 20 79 years. Bone mineral density (BMD) was determined using dual-energy X-ray absorptiometry (DXA) on Hologic bone densitometers (Hologic, Waltham, Mass.). Most of the subjects were examined with a QDR 4500. The BMD of the lumbar vertebrae was virtually constant between 20 and 49 years (test for trend: P=0.66); the BMD values between 20-45 in premenopausal women (mean 1.036; SD 0.109 g/cm(2)) were thus defined as the peak bone mass values, significantly lower compared to the Hologic reference curve (mean 1.079, SD 0.11 g/cm(2)). The mean BMD values of the femoral neck were virtually identical to those of the NHANES study in the first 3 decades; after the age of 50 the BMD values were slightly greater than those of the NHANES subject. The subject classification according to the WHO criteria was similar using the DINS and NHANES reference values for the femur; for the spine, the Hologic reference values classified a larger proportion of women as osteoporotic (21 vs. 16%) or osteopenic (42 vs. 38%) compared to DINS. PMID- 14530831 TI - Sacral neuromodulation for the treatment of refractory interstitial cystitis: outcomes based on technique. AB - Patients with refractory interstitial cystitis (IC) underwent testing with sacral nerve modulation via either a traditional percutaneous approach or a staged procedure. Implanted patients were followed with scaled questionnaires and voiding diaries. Twenty-six patients who had a permanent InterStim placed had a reduction in 24-h voids of 51%. More than two-thirds of patients reported a moderate or marked improvement in urinary frequency, urgency, pelvic pain, pelvic pressure, incontinence and overall quality of life. The test to implant rate of a traditional percutaneous procedure was 52%, compared to a staged procedure of 94%. Assessing sensory response at the time of implant reduced the reoperation rate from 43% to 0%. Ninety-six per cent stated they would undergo an implant again and recommend the therapy to a friend. We concluded that sacral nerve modulation can treat refractory IC symptoms. The response to therapy and the reoperation rate are dependent on the technique used to test and implant the device. PMID- 14530832 TI - A randomized controlled trial comparing a modified Burch procedure and a suburethral sling: long-term follow-up. AB - The aim of this study was to compare the long-term results of a modified Burch procedure with a sling procedure for the treatment of stress incontinence with a low-pressure urethra. Thirty-six women with urodynamic stress incontinence, low pressure urethra, urethral hypermobility and no significant pelvic organ prolapse were randomly assigned to undergo either a modified Burch procedure ( n=19) or a sling placement ( n=17). Cure of the stress incontinence (defined as a negative stress test and negative pad-weight test) was the primary long-term endpoint. Secondary endpoints included subjective cure of stress incontinence (defined as no incontinence episodes on a 1-week voiding diary) and voiding function studies. Comparisons of group means were performed with the Mann-Whitney U-test, pooled variance t-tests and separate variance t-tests. Proportions were compared with Fisher's exact test. A logistic regression analysis was performed to control for covariates that differed in our two groups despite randomization. Long-term follow-up (mean=72.6 months) was available for 82% (28/34) of the original study group. The objective cure rates for the Burch and sling groups were 84.6% and 100%, respectively ( P=0.17). Mean uroflowmetry rates for the Burch and sling groups were 7.38 and 6.8 ml/s, respectively ( P=0.58, 95%CI -2.5, 4.4). Mean postvoid residual volumes for both groups were 35 ml ( P=0.97, 95% CI -23.8, 65.9). Two sling patients (12%) required partial resection of their slings because of erosion. Both patients remained continent. In terms of voiding function and stress incontinence cure, there were no differences between groups undergoing modified Burch or sling procedures for treatment of urodynamic stress incontinence with low-pressure urethra. PMID- 14530833 TI - The effect of pelvic floor training on sexual function of treated patients. AB - The aim of this study was to determine the effects of improvements in urinary incontinence resulting from pelvic floor rehabilitation on the sexual function of patients. The study involved 42 clinic patients who received pelvic floor rehabilitation treatment. Their sexual histories were obtained through face-to face interviews. Pelvic muscle strength was measured with a perineometer. Improvement in incontinence was measured with the pad test. Seventeen women reported decreased sexual desire before the treatment; 5 of these indicated improvement after treatment. Nine of 17 women who experienced dyspareunia prior to treatment reported an improvement afterwards, and four women reported complete relief from pain. Five of 15 women who complained of difficulty in reaching climax before the treatment experienced improvement in this area. In conclusion, an improvement in sexual desire, performance during coitus and achievement of orgasm were observed in women who received pelvic floor muscle rehabilitation. No change was seen in the arousal and resolution stages of sexual activity. PMID- 14530834 TI - Mechanical properties of urogynecologic implant materials. AB - Synthetic suburethral slings have recently become popular despite the risk of erosion commonly associated with synthetic implants. Some of these materials seem to have unexpectedly low erosion rates. Based on the hypothesis that erosion is due, in part, to biomechanical properties, we undertook an in vitro study. The biomechanical properties of eight non-resorbable synthetic implant materials, stiffness (slope, N/mm) and peak load (N) were determined from load vs. displacement curves. Open-weave Prolene mesh showed unique biomechanical properties compared to other tested materials. The tension- free vaginal tape had the lowest initial stiffness (0.23 N/mm), i.e. low resistance to deformation at forces below the elastic limit, whereas the stiffest implant tested, a nylon tape, reached 6.83 N/mm. We concluded that the TVT and other wide-weave Prolene tapes have unique biomechanical characteristics. These properties may be at least partly responsible for the apparent clinical success of the implants. PMID- 14530835 TI - Anorectal symptoms after various modes of vaginal delivery. AB - The aim of this study was to determine the prevalence of, and the changes in, anorectal symptoms following different modes of vaginal delivery in primiparous women. Six hundred and seventeen questionnaires were distributed to primiparous women 3-4 years after delivery. The questionnaires were designed to obtain information regarding the development of anorectal symptoms, including the type of symptoms experienced, their severity and their impact on lifestyle. A total of 479 questionnaires were returned, representing a response rate of 77.6%. Women included in the study were divided into three groups on the basis of the mode of delivery (normal vaginal, vacuum extraction and forceps). Any episode of fecal incontinence was considered to be abnormal. Following delivery, de novo incontinence developed in 22%. There was no significant difference between the three modes of vaginal delivery in terms of the development of fecal incontinence (normal vaginal delivery 22%, vacuum extraction 20%, forceps delivery 26%). Furthermore, analysis of obstetric variables could not identify one significant independent risk factor for anorectal incontinence. The results of this study suggest that instrumental vaginal deliveries are as safe as a normal vaginal delivery in terms of the development of anorectal symptoms. PMID- 14530836 TI - Do Asian women have less pelvic organ mobility than Caucasians? AB - Epidemiological studies have postulated racial differences in the incidence and prevalence of pelvic floor disorders. There are anecdotal data from cadaver dissections suggesting that Asian women benefit from stronger pelvic support structures. A prospective observational clinical study was conducted in order to test for differences in pelvic organ support in 200 nulliparous pregnant women. Assessment included translabial ultrasound, documenting the position of the pelvic organs and mobility on Valsalva relative to the inferoposterior symphyseal margin. The largest ethnic groups were Asian ( n=16) and Caucasian women ( n=161). On comparing the groups, both antepartum and postpartum analyses showed significantly less pelvic organ mobility in Asians. This was true for virtually all parameters of organ mobility and both anterior ( P=0.002 antepartum, P=0.009 postpartum) and posterior compartments ( P=0.04 antepartum, P=0.02 postpartum). No significant differences were detected for cervical mobility. It was concluded that Asian women seem to show less mobility of the anterior and posterior vaginal compartments than Caucasians. PMID- 14530837 TI - Anorexia nervosa and pelvic floor dysfunction. AB - Pelvic floor dysfunction in women with eating disorders is an underexplored area. We present a case of pelvic floor dysfunction in a nulliparous woman with anorexia nervosa. PMID- 14530838 TI - One-year urodynamic outcome and quality of life in patients with concomitant tension-free vaginal tape during pelvic floor reconstruction surgery for genitourinary prolapse and urodynamic stress incontinence. AB - The aim of the study was to examine the 1-year urodynamic outcome and quality of life in patients who have had concomitant tension-free vaginal tape insertion during pelvic floor reconstruction surgery. The medical notes of a retrospective cohort of 45 patients who had undergone tension-free vaginal tape together with pelvic floor reconstruction surgery were reviewed. The operative information, the results of the urodynamic studies and the change in the quality-of-life scores 1 year after surgery were examined. The quality of life was assessed with both general and disease-specific quality-of-life questionnaires (General Health Questionnaire (GHQ-12), Urogenital Distress Inventory (UDI-6), Incontinence Impact Questionnaire (IIQ-7)). Patient satisfaction was assessed with the validated Chinese version of the Client Satisfaction Questionnaire (CSQ). Forty five patients underwent tension-free vaginal tape insertion together with pelvic floor reconstruction surgery. The overall objective cure rate was 43%. There was a significant improvement in the disease specific quality-of-life assessment (UDI 6 score 38.3-15.5; P<0.01) and (IIQ-7 score 15.0-4.0; P<0.01). The patients who had a concomitant cystocele repair had a worse objective cure rate than patients without concomitant cystocele repair (38% vs 67%; P=0.19). PMID- 14530839 TI - Rehabilitation of the short pelvic floor. I: Background and patient evaluation. AB - Pelvic floor physical therapists have traditionally focused on rehabilitation of the weak pelvic floor of normal length. With the recognition that many urogynecologic symptoms arise from the presence of a short, painful pelvic floor, the role of the physical therapist is expanding. Clinically, the pelvic floor musculature is found to be short, tender, and therefore weak. There are associated trigger points and characteristic extrapelvic connective tissue abnormalities. We report the characteristic patterns of myofascial and connective tissue abnormalities in 49 patients presenting with this syndrome. PMID- 14530840 TI - Rehabilitation of the short pelvic floor. II: Treatment of the patient with the short pelvic floor. AB - Several urogynecologic syndromes are associated with the clinical finding of a short, painful, tender and weak pelvic floor and a variety of connective tissue abnormalities. Techniques for rehabilitation include the avoidance of perpetuating factors, rehabilitation of extrapelvic musculoskeletal abnormalities, the use of manual techniques and needling to promote resolution of connective tissue problems, closure of any diastasis recti, and transvaginal/transrectal manual release of muscular trigger points and contractures. Therapy can be facilitated by pudendal or epidural nerve block. Patients contribute to their success through home maintenance programs. PMID- 14530841 TI - Expression of estrogen receptor-alpha and -beta in anterior vaginal walls of genuine stress incontinent women. AB - Our objective was to study the expression of estrogen receptor (ER) isoforms, ER alpha and ER-beta, in the anterior vaginal wall of menopausal and fertile women with genuine stress incontinence (SI) by immunohistochemistry and Western blot analysis. Eighteen menopausal women with SI who either were or were not taking estrogen/progestin replacement therapy and 14 fertile women with SI who either were or were not taking contraceptives were enrolled in the study. Biopsies from the suburethral anterior vaginal wall were obtained at tension-free vaginal tape (TVT) operation. Monoclonal antibody to ER-alpha and polyclonal antibody to ER beta were used to stain frozen sections of vaginal tissue. The receptor expressions were scored based on percentage of positive cells. ER-alpha was detected in vaginal epithelial, stromal and smooth muscle cells. In menopausal SI women ER-alpha was detected significantly more frequently in the vaginal walls of estrogen/progestin-treated patients than in those who were untreated. Fertile SI women had significantly higher expression of ER-alpha than menopausal SI women. ER-alpha was not observed in vaginal blood vessels. ER-beta was detected in epithelial and vascular smooth muscle cells of the vagina. No significant difference in ER-beta expression was observed between different groups of patients. The expression of ER-alpha was not correlated with that of ER-beta. Both ER-alpha and -beta were detected, indicating a potential role for both types of estrogen receptor in the human vaginal wall. The expression of ER-alpha, but not of ER-beta, in menopausal SI women was regulated by estrogen/progestin replacement therapy. The presence of ER-beta in vaginal vascular smooth muscle cells raises the possibility of vascular effects of estrogen on the human vaginal wall. PMID- 14530842 TI - Utilization of preoperative urodynamic investigations by gynecologists who frequently operate for female urinary incontinence. AB - Our objective was to determine the availability and utilization of urodynamic investigations by gynecologists in the preoperative evaluation of women with urinary incontinence. Gynecologists in the UK, USA, Australia, New Zealand and Canada were asked what urodynamic investigations were required for four clinical scenarios. Analysis was restricted to frequent operators. Urodynamic investigations were available to 70% of frequent operators. For uncomplicated stress incontinence, cystometry was utilized by 72% of subspecialists and 44% of generalists ( P<0.001) who had access to urodynamic investigations. For stress incontinence and straining to void, uroflowmetry was utilized by 73% of subspecialists and 46% of generalists ( P<0.001) who had access to urodynamic investigations. We concluded that many gynecologists who frequently operate for female urinary incontinence do not have access to urodynamic investigations or do not utilize urodynamic investigations, or utilize investigations in a way that may be inadequate for the clinical problem. There are differences in utilization between subspecialists and generalists that are not explained by access. These observations could be explained by poor understanding or a lack of belief in the value of urodynamic investigations. PMID- 14530843 TI - Reproducibility of grading scales of pelvic muscle strength: standard versus tampon scale. AB - The purpose of this study was to demonstrate the reproducibility of an objective scale utilizing a tampon to assess pelvic muscle strength and to compare it to a digital evaluation of muscle strength [1]. Thirty women presenting to the office were enrolled in the study. Their pelvic floor muscle strength was separately assessed by both examiners using both scales. The reproducibility of the scales was then analyzed by Cohen's kappa measure of interrater agreement. The tampon scale was found to have a measure of agreement of 0.76 (excellent interrater agreement if >0.75). In contrast, the digital scale was found to have a measure of agreement of 0.68 (fair agreement if 0.40-0.75). Therefore, the tampon scale is an easy and reproducible approach for assessing pelvic floor muscle strength. PMID- 14530844 TI - Necrotizing fasciitis after tension-free vaginal tape (TVT) placement. AB - A 62-year-old homemaker underwent an uneventful TVT (tension-free vaginal tape) procedure. Eleven days postoperatively she presented to the Emergency Department with agonizing low abdominal pain, elbow pain, fever, and drainage from her suprapubic puncture sites. Her evaluation was consistent with a diagnosis of necrotizing fasciitis and she was started on antibiotics and taken to surgery for exploration and debridement, where the diagnosis was confirmed. This is the first report of necrotizing fasciitis following TVT placement. PMID- 14530845 TI - Back injuries and pain in adolescents attending a ski high school. AB - This study compared overuse injuries, small abnormalities, and pain alone in different types of skiing and activity levels. Subjects were 45 ski high school athletes aged 15-19 years. We found the back and knees significantly more prone to activity-related injuries and pain than other body regions. Thirty subjects (67%) had low back pain on the first examination, and 17 had pain caused by overuse of the back. This was more frequent among active, young competitive boys in the cross-country skiing group, with mature height less than 99%, than among noncompetitive boys in the same group or among the alpine skiers. Neck pain was reported only by girls. Eight subjects had low back pain related to small abnormalities. The subjects were given counseling about training and physiotherapy. The 1-year follow-up found a significant reduction in back pain due to overuse injuries (from 17 to 3 subjects) and indistinct neck pain (from 10 to 2) but no reduction in pain in those with small abnormalities in the low back. Low back pain was common in young athletes, particularly in cross-country skiers. Excessively rapid progression of training and faulty technique increased back pain complaints from 36% when entering the school to 67% at the first examination. A proper evaluation and treatment of overuse injuries and accurate counseling of training types, volume, and progression reduced the low back pain problems to 29% of subjects 1 year later in spite of a maintained high activity level. A minimum entrance requirement to conditioning seems mandatory for students attending a ski high school. PMID- 14530846 TI - A computer assisted surgical technique for retrograde autologous osteochondral grafting in talar osteochondritis dissecans (OCD): a cadaveric study. AB - This study describes a new method for the treatment of osteochondritis dissecans (OCD) in the medial talar dome. Ten cadaveric lower extremities were used to develop and evaluate a retrograde osteochondral grafting technique applying computer-assisted surgery. With the help of a computed tomography (CT)-based navigation system, a guide wire was placed from the lateral talar process into the posteromedial talar trochlea where OCD lesions are predominantly located. Cannulated reamers and arthroscopic shavers were used for the preparation of the recipient hole. The grafts, with diameters of 4.5 mm, 6.5 mm or 8.5 mm were harvested from the lateral femoral trochlea and inserted in a retrograde fashion. The last five cadavers were analyzed for accuracy of surface reconstruction and graft stability. For this purpose a medial malleolar osteotomy and a CT scan was performed. We found steps in the joint surface to range from 0.5 mm to 1.5 mm (mean 0.9 mm, SD 0.4) with the graft always below the surrounding cartilage. Graft subsidence occurred at an applied force of 26.4 +/- 4.6 N. This study indicates that osteochondral cylinders can be inserted in a retrograde fashion to reconstruct the posteromedial talus. Good surface congruency and sufficient graft stability can be achieved. PMID- 14530847 TI - Percutaneous Achilles tendon repair with and without endoscopic control. AB - One hundred and fifty six patients were treated using the modified double suture technique for percutaneous Achilles tendon repair between 1994 and 1998. Endoscopy was used in 67 cases. The first ten cases were dropped (learning curve), 57 were followed (E-group). Percutaneous suture without endoscopy was performed in 89 patients. Two could not be followed (went abroad), so this group consists of 87 patients (P-group). Mean age: E-group 37.8 (22-60) years, P-group 38.9 (20-68) years. Male-female ratio: E 49/8, P 74/13. There were 54 and 83 athletes in groups E and P respectively. Follow-up period was 12-60 months. Overall re-rupture rate was 6/144 (4.2%). Two total and 3 partial re-ruptures were in the P-group, and 1 partial was in the E-group. Fusiform thickening of the tendon (delayed healing) occurred in 4 cases in each group. The mean plantar flexion strength compared with the non-affected side was 89% in the P-group and 86% in the E-group. The length of time before returning to sports activity ranged from 4 to 6 months after surgery in both groups. Subjective results were excellent to good in 88% (P-group) and in 89% (E-group) of the cases. On the basis of the results, the percutaneous double suture technique proved to be a simple and safe method for Achilles tendon repair with or without the use of an endoscope. The re-rupture rate was lower in the endoscopic controlled group. The basic goal of the endoscopy was to control the adaptation of the tendon ends. This method yielded further operative possibilities and benefits as well. PMID- 14530848 TI - Arthroscopic removal of the osteoid osteoma on the neck of the talus. AB - Juxta-articular osteoid osteomas arising around the ankle are unusual. Tumors arising on the neck of the talus will commonly produce symptoms mimicking monoarticular arthritis or trauma. Patients are usually treated for arthritis or ankle sprain, which often leads to a delay in definitive diagnosis. We present an arthroscopic removal of an osteoid osteoma on the neck of talus, and review the literature. PMID- 14530849 TI - The cartilaginous and osseous geometry of the femoral trochlear groove. AB - Photography was used to study the geometry of the cartilaginous and osseous contours of the distal femur and the orientation of the trochlear groove in 9 fresh-frozen and 24 embalmed knees. The sulcus angle (146.1 degrees +/-5.5 degrees ) decreased from 0 degrees to 50 degrees of femoral flexion then increased afterwards. The maximum slope of the lateral femoral condyle (20.2 degrees +/-5.2 degrees ) also decreased with flexion. Both the sulcus angle ( p=0.0007) and maximum slope ( p=0.0001) were larger at 0 degrees than they were for 60 degrees cartilaginous surfaces. The lateral femoral condylar height decreased, whilst the medial femoral condylar height increased as the flexion increased. The femoral groove was midway between the two femoral epicondyles (49.5+/-3.9%), but deviated laterally as the flexion angle increased. The groove axis deviated distally and laterally from the femoral anatomical axis for both cartilaginous and bony surfaces, and the angle between the groove and anatomical axes was similar for both cartilaginous (19.1 degrees ) and osseous (16.8 degrees ) surfaces. Articular cartilage is not well represented on radiography yet it had a significant effect on the distal femoral geometry, and should be taken into account when evaluating the patellofemoral joint. PMID- 14530850 TI - Avulsion fractures of the tibial tuberosity in adolescent athletes treated by internal fixation and tension band wiring. AB - Avulsion of the tibial tuberosity is a rarely reported fracture. It is mainly considered as an athletic injury accounting for less than 3% of all epiphyseal lesions. In this study, we hypothesized that the use of tension band wiring as a supplement of the internal fixation for the avulsion fractures of the tibial tuberosity would lead the adolescent athletes to a more effective rehabilitation program and an earlier resumption of their previous activity level. Ten patients were treated in our department over a period of 11 years (1985-1995). Operative treatment was thought necessary for all our cases due to tibial tuberosity displacement. Open reduction and internal fixation in combination with tension band wiring was used. The result in all cases was that the reduction was maintained intact and the fracture united. The functional results were excellent, and all patients returned to their previous athletic activities. Our conclusion is that the combination of internal fixation and tension band wiring for avulsion fractures of the tibial tuberosity seems to be more effective and advantageous than conservative or other surgical methods. Avoiding the need of external support and allowing early joint motion, the method described prevents serious quadriceps atrophy, allowing the young athletes to return earlier to their previous sport activities. PMID- 14530851 TI - The effects of anterior cruciate ligament reconstruction on tibial rotation during pivoting after descending stairs. AB - Recent in vitro research suggests that ACL reconstruction does not restore tibial rotation. This study investigated rotational knee joint stability in vivo during a combined descending and pivoting movement that applies a high rotational load to the knee joint. We studied 20 ACL reconstructed patients (bone-patellar tendon bone graft) and 15 matched controls with a six-camera optoelectronic system performing the examined movement. In the control group the results showed no significant differences in the amount of tibial rotation between the two sides. No significant differences were also found between the contralateral intact leg of the ACL group and the healthy control. However, a significant difference was found within the ACL reconstructed group and between the reconstructed and the contralateral intact leg. Therefore ACL reconstruction may not restore tibial rotation even though anterior tibial translation has been reestablished. PMID- 14530852 TI - Magnetic resonance imaging of anterior cruciate ligament tears: reevaluation of quantitative parameters and imaging findings including a simplified method for measuring the anterior cruciate ligament angle. AB - We evaluated the diagnostic utility of magnetic resonance imaging (MRI) for predicting anterior cruciate ligament (ACL) tears using both quantitative parameters and nonquantitative imaging findings. MRI examinations were retrospectively evaluated in a group of patients with arthroscopically confirmed complete ACL tear and in a control group with arthroscopically confirmed intact ACL. We evaluated multiple MRI features to compare their sensitivity and specificity for detecting ACL tears. Particular emphasis is put on the evaluation of three different quantitative parameters, including a simplified method for measuring the ACL angle. With a threshold value of 45 degrees the ACL angle reached a sensitivity and specificity of 100% for detecting ACL tears. With a threshold value of 0 degrees the Blumensaat angle had a sensitivity of 90% and a specificity of 98%. Finally, a threshold value of 115 degrees gave the posterior cruciate ligament angle a sensitivity of 70% and a specificity of 82%. Discontinuity was found to be the most useful of the ACL abnormalities. Of the secondary findings anterior tibial displacement was the best predictor of ligamentous injury. However, ACL abnormalities and secondary findings, alone or combined, failed to surpass the diagnostic value of the ACL angle for predicting ACL tears. Quantitative parameters are thus good predictors of ACL tears and may increase the overall sensitivity and specificity of MRI. The ACL angle may be confidently measured in a single MRI section and can be considered to be the most reliable quantitative parameter for detecting ACL tears. PMID- 14530853 TI - The relationship between isokinetic quadriceps strength and laxity on gait analysis parameters in anterior cruciate ligament reconstructed knees. AB - Gait alterations after ACL reconstruction have been reported in the literature. The current study examined a group of 14 patients who all had an ACL reconstruction with a patellar tendon autograft. Kinetic and kinematic data were obtained from the knee during walking. The flexion-extension deficit (FED) calculated from the angular difference between maximal flexion and maximal extension during the stance phase in the ACL-reconstructed and the normal knee was measured. We investigated whether these alterations in gait are related to quadriceps strength and residual laxity of the knee. It may be that patients modify their gait patterns to protect the knee from excessive anterior translation of the tibia by reducing the amount of extension during stance. On the other hand, persistent quadriceps weakness may also cause changes in gait patterns as the quadriceps is functioning as an important dynamic stabilizer of the knee during stance. Results showed that patients had a significantly higher FED value (4.9+/-4.0) than a healthy control group in a previous study (1.3+/ 0.9). This is caused mainly by an extension deficit during midstance. External extension moments of the knee (TZMAX were significantly lower in the current patients group than in a healthy control group (TZMAX -0.27+/-0.19 Nm/kg in patients vs. -0.08+/-0.06 Nm/kg in controls). There were no significant correlations between quadriceps strength and gait analysis parameters. Furthermore no correlation was found between the amount of laxity of the knee and gait. The relevance of this study lies in the fact that apparently the measured gait alterations cannot be explained solely by often used biomechanical indicators such as laxity and strength. The measured gait alterations may be a result of the surgical procedure with subsequent modified motor programming. PMID- 14530854 TI - Fracture of the femoral component of a Brigham unicompartmental knee: a case report. AB - We report the failure of a femoral component in a unicompartmental knee replacement 6 years after implantation. The implant fractured 15 mm from the anterior tip, and scanning electron microscopy confirmed fatigue to be the cause of failure. The clinical and laboratory findings are presented, and the causes and implications of this mode of implant failure are discussed. PMID- 14530855 TI - Trends in organ preservation. AB - Organ preservation aims to provide a viable graft with primary function post transplant. The current basis of preservation for transplantation is static cold storage using specific preservation solutions which minimise cellular swelling and membrane pump activity, thus maintaining cellular ATP levels. The current organ shortage and consequent expansion of donor criteria places even greater reliance on minimising graft injury during preservation. This review focuses on current and future advances in preservation technology. The key areas of advance are additives to preservation solutions, alternatives/adjuncts to preservation solutions including perfluorocarbons. A major area of advance is in the modulation of organs during the storage period. This may be achieved by biochemical additives or genetic manipulation. Machine perfusion technology is improving, and this is discussed together with the recent concept of warm (normothermic) perfusion as an alternative means of preservation. The authors provide an overview over the current methods of organ preservation. Cold storage, effective in the short-term is insufficient for marginal organs, does not allow assessment of viability markers, and provokes ischaemic injury. Potential strategies for minimising ischaemic injury include additives to preservation solutions; the two-layer method with perfluorcarbons and UW solution-at present limited to pancreas preservation; organ modulation; organ preconditioning and genetic modification of organs. In particular, the authors illuminate the potential in a reappraisal of the concept of normothermic perfusion. PMID- 14530856 TI - Elevated troponin T concentrations in critically ill patients. AB - OBJECTIVE: To determine the incidence of troponin T elevations among a selected group of critically ill patients, to correlate these findings to electrocardiographs, and to compare troponin T-positive and T-negative patients in relation to clinical parameters. DESIGN: Prospective study. SETTING: Mixed surgical and medical intensive care unit. PATIENTS: Thirty-four consecutive critically ill patients who were mechanically ventilated or underwent thoracic or vascular surgery. INTERVENTIONS: Blood samples were collected at admission, the next morning, and 24 h after the second blood sampling. These samples were used for troponin T measurement and electrocardiographs were made when troponin T levels were elevated. MAIN RESULTS: Eleven of 34 patients (32%) had elevated troponin T levels, which were already present upon admission in eight out of 11 patients (73%). Most patients underwent surgery prior to ICU admission (21 of 34 patients). Significantly ( P=0.0055) more troponin T-positive patients underwent acute surgery, and significantly more ( P=0.045) troponin T-positive patients suffered from hypotension. Only four of the troponin T-positive patients were diagnosed as suffering from an acute myocardial infarction based on electrocardiographs. All troponin T-positive patients had coronary artery disease: nine had a history of CAD and two had actual CAD. No difference in mortality rates was observed between troponin T-positive and T-negative patients. CONCLUSIONS: An unexpectedly high percentage of included patients had troponin T elevations, which could be corroborated by electrocardiographs in only four cases suggesting that a high percentage of critically ill patients with a history of CAD suffer from clinically unrecognised (minor) myocardial damage. PMID- 14530857 TI - Decisions to forgo life-sustaining therapy in ICU patients independently predict hospital death. AB - OBJECTIVE: More than one-half the deaths of patients admitted to intensive care units (ICUs) occur after a decision to forgo life-sustaining therapy (DFLST). Although DFLSTs typically occur in patients with severe comorbidities and intractable acute medical disorders, other factors may influence the likelihood of DFLSTs. The objectives of this study were to describe the factors and mortality associated with DFLSTs and to evaluate the potential independent impact of DFLSTs on hospital mortality. DESIGN AND SETTING: Prospective multicenter 2 year study in six ICUs in France. PATIENTS: The 1,698 patients admitted to the participating ICUs during the study period, including 295 (17.4%) with DFLSTs. MEASUREMENTS AND RESULTS: The impact of DFLSTs on hospital mortality was evaluated using a model that incorporates changes in daily logistic organ dysfunction scores during the first ICU week. Univariate predictors of death included demographic factors (age, gender), comorbidities, reasons for ICU admission, severity scores at ICU admission, and DFLSTs. In a stepwise Cox model five variables independently predicted mortality: good chronic health status (hazard ratio, 0.479), SAPS II score higher than 39 (2.05), chronic liver disease (1.463), daily logistic organ dysfunction score (1.357 per point), and DFLSTs (1.887). CONCLUSIONS: DFLSTs remain independently associated with death after adjusting on comorbidities and severity at ICU admission and within the first ICU week. This highlights the need for further clarifying the many determinants of DFLSTs and for routinely collecting DFLSTs in studies with survival as the outcome variable of interest. PMID- 14530858 TI - Intra-abdominal pressure may be decreased non-invasively by continuous negative extra-abdominal pressure (NEXAP). AB - OBJECTIVE: To investigate the possibility of artificially decreasing intra abdominal pressure (IAP) by applying continuous negative pressure around the abdomen. MATERIAL AND METHODS: We investigated the effects of negative extra abdominal pressure (NEXAP) on IAP and central venous pressure (CVP) in 30 patients admitted to our intensive care unit (age 57+/-17 years, BMI 26.1+/-4.0 kg/m2, SAPS II 41.8+/-17.0). Patients with severe hemodynamic instability and/or those admitted following a laparotomy were not studied. Measurements included bladder pressure as an estimate of IAP, CVP, invasive mean arterial pressure (MAP) and heart rate (HR). In five patients extensive hemodynamic measurements were also taken using a Swan-Ganz catheter. Following measurements at baseline (Basal), NEXAP (Life Care - Nev 100, Respironics) was applied on the abdomen, in random order, at a pressure equal to IAP (NEXAP0), 5 cmH(2)O (NEXAP-5) or 10 cmH(2)O (NEXAP-10) more negative than NEXAP0. RESULTS: Basal IAP ranged from 4 to 22 mmHg. NEXAP decreased IAP from 8.7+/-4.3 mmHg to 6+/-4.2 (Basal vs NEXAP0 p<0.001). There was a further decrease of IAP when more negative pressure was applied: 4.3+/-3.2 mmHg, 3.8+/-3.7 mmHg (NEXAP-5 and NEXAP-10 vs NEXAP0, respectively, p<0.001). Similarly, CVP decreased from 9.3+/-3.4 mmHg to 7.5+/-3.8 (Basal vs NEXAP-10, p<0.001). The lower the IAP when NEXAP was applied, the lower the CVP (r2=0.778, p<0.001, multiple linear regression). When measured, cardiac output did not significantly change with NEXAP. CONCLUSIONS: Negative extra abdominal pressure may be applied in critically ill patients to decrease intra abdominal pressure non-invasively. PMID- 14530859 TI - Intravenous sildenafil and inhaled nitric oxide: a randomised trial in infants after cardiac surgery. AB - OBJECTIVE: To investigate the acute effects of intravenous sildenafil on haemodynamics and oxygenation, and its interaction with inhaled nitric oxide (iNO) in infants at risk of pulmonary hypertension early after cardiac surgery. DESIGN: Prospective, randomised trial. SETTING: Paediatric intensive care unit of a children's hospital. PATIENTS AND PARTICIPANTS: Sixteen ventilated infants early after closure of ventricular or atrioventricular septal defects, were randomly assigned to one of two groups. The study was completed in 15 infants. INTERVENTIONS: Studies were commenced within 7 h of separation from bypass. Seven infants received iNO (20 ppm) first, with the addition of intravenous sildenafil (0.35 mg/kg over 20 min) after 20 min. Eight infants received sildenafil first, iNO was added after 20 min. Vascular pressures, cardiac output and a blood gas were recorded at 0, 20 and 40 min. MEASUREMENTS AND RESULTS: In infants receiving iNO first, iNO lowered the pulmonary vascular resistance index (PVRI) from 3.45 to 2.95 units (p=0.01); sildenafil further reduced PVRI to 2.45 units p<0.05). In those receiving sildenafil first, PVRI was reduced from 2.84 to 2.35 units (p<0.05) with sildenafil, and fell to 2.15 units (p=0.01) with the addition of iNO. In both groups, sildenafil reduced the systemic blood pressure and systemic vascular resistance (p<0.01) and worsened arterial oxygenation and the alveolar arterial gradient (p<0.05). CONCLUSION: Intravenous sildenafil augmented the pulmonary vasodilator effects of iNO in infants early after cardiac surgery. However, sildenafil produced systemic hypotension and impaired oxygenation, which was not improved by iNO. PMID- 14530860 TI - Measurement of lung volume in mechanically ventilated monkeys with an ultrasonic flow meter and the nitrogen washout method. AB - OBJECTIVE: Measurement of functional residual capacity (FRC) during mechanical ventilation is important to standardise respiratory system compliance and adjust the ventilator settings to optimise lung recruitment. In the present study we compared three methods to measure FRC. DESIGN: The bias flow nitrogen washout technique (FRC(N2MC)), the multiple breath nitrogen washout (FRC(MBNW)) and the multiple breath sulphur-hexafluoride washout using the molar mass signal of an ultrasonic flow meter (FRC(MBSF6)) were compared in six adult monkeys after endotracheal intubation and during spontaneous breathing and mechanical ventilation at three different positive end-expiratory pressure (PEEP) levels of 0, 5 and 10 cmH2O. SETTING: Animal research laboratory. RESULTS: We found good agreement between all three methods and they all accurately measured changes in FRC when PEEP was increased. The coefficients of variance of the three measurement techniques were in the same range (1.3-9.2%). CONCLUSION: The measurement of the tracer gas concentration with the molar mass signal of the ultrasonic flow meter provides a good and simple alternative to respiratory mass spectrometer for FRC measurements in ventilated subjects. PMID- 14530861 TI - The FOXC2 -512C>T variant is associated with hypertriglyceridaemia and increased serum C-peptide in Danish Caucasian glucose-tolerant subjects. AB - AIMS/HYPOTHESIS: The transcription factor FOXC2 plays a key role in adipocyte differentiation and the FOXC2 gene is a candidate gene for Type 2 diabetes, obesity and dyslipidaemia. We investigated whether the FOXC2 -512C>T promoter variant is associated with Type 2 diabetes or its intermediary phenotypes in glucose tolerant subjects. METHODS: The variant was genotyped using PCR-RFLP in 705 unrelated Type 2 diabetic patients, 505 unrelated glucose-tolerant control subjects and 219 glucose-tolerant offspring of Type 2 diabetic probands. RESULTS: The frequency of the T-allele was 58% (95% CI 56-61%) and 59% (56-62%) among the Type 2 diabetic patients and the unrelated glucose-tolerant control subjects, respectively ( p=0.6). Among the glucose-tolerant subjects, the T-allele carriers had higher fasting serum triglyceride ( p=0.03), fasting serum C-peptide concentrations ( p=0.009) and insulinogenic index ( p=0.04). Furthermore, in glucose-tolerant women, the waist-to-hip ratio was significantly higher in carriers of the T-allele. CONCLUSION/INTERPRETATION: Our data suggest that the FOXC2 -512C>T variant is not associated with Type 2 diabetes. However, among glucose-tolerant subjects the variant is associated with hypertriglyceridaemia and increased fasting serum C-peptide. PMID- 14530862 TI - Associations between hyperglycaemia and somatic transversion mutations in mitochondrial DNA of people with diabetes mellitus. AB - AIMS/HYPOTHESIS: Considering that increased oxidative stress induced by hyperglycaemia plays a possible role in the pathogenesis of diabetic complications and that mitochondrial DNA (mDNA) is thought to be more vulnerable than nuclear DNA, we investigated what somatic mutations actually occur in the mDNA of diabetic patients. We also studied the relations between those mutations and urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) which is known to increase considerably in people with diabetes. METHODS: We identified somatic mutations by subcloning and sequencing two segments of mDNA [control region (nt 15996-16401) and the segment encompassing t-RNA(Leu(UUR))(nt 3149-3404)] in the peripheral blood cells of six diabetic women and control subjects matched for age and sex. This was done in 20 colonies each. In each case we also assayed urinary 8-OHdG. RESULTS: No difference in the aggregate somatic mutational burden of mDNA was found between patients and control subjects. However, the incidence of somatic transversion mutations in mDNA was significantly higher in diabetic patients than in control subjects (13.93+/-4.57 x 10(-5) vs 1.27+/-1.27 x 10(-5) mutations per base pair; p=0.031, according to Mann-Whitney U-test). There was no significant difference in transition mutations. A correlation was found between the transversion mutational burden and HbA(1)c values, but not between it and 8-OHdG content in the urine. CONCLUSIONS/INTERPRETATION: We showed that somatic transversion point mutations of mDNA increase in diabetic patients. Such transversion mutations can become a new biomarker for mDNA damage associated with hyperglycaemia and possibly caused by oxidative stress but not reflected by urinary 8-OHdG. PMID- 14530863 TI - Association analysis of the SHC1 gene locus with longevity in the Japanese population. AB - The SHC1 gene encodes a signaling and transforming protein that has been implicated in the aging process in worms and mammals. In this study we examined 230 Japanese centenarians and 180 healthy younger controls and looked at the SHC1 locus as a candidate region that may be associated with longevity. We identified 12 single nucleotide polymorphisms (SNPs) within a 10-kb region encompassing the entire SHC1 gene from the DNA of 30 centenarians and 24 healthy younger controls. Five SNPs, including three nonsynonymous sites, lay within coding elements, six were located within introns, and one was in the 3' untranslated region. All of these SNPs were relatively rare, with a minor allele frequency of less than 5% in our subjects. A pairwise linkage disequilibrium analysis using the r2 statistic showed that two of the SNP pairs are in tight linkage disequilibrium at this locus. We investigated the possible association of SHC1 with longevity using association analyses with allelotypes and haplotypes but found that the SNPs identified in SHC1 had no impact on longevity for Japanese centenarians. PMID- 14530864 TI - Comparison of Agrobacterium-mediated transformation of four barley cultivars using the GFP and GUS reporter genes. AB - Experiments were conducted to produce transgenic barley plants following infection of immature embryos with Agrobacterium tumefaciens. Transformed callus was obtained using hygromycin resistance as a selectable marker and either green fluorescent protein (GFP) or beta-glucuronidase (GUS) as a reporter. Significantly reduced plant transformation frequencies were obtained with the GFP gene compared to GUS. However, GFP proved to be an excellent reporter of early transformation events and was used to compare four barley cultivars for efficiency in two phases of transformation: the generation of stably transformed barley callus and the regeneration of plantlets from transformed callus. Transformed callus was generated at a high frequency (47-76%) in all four cultivars. Regeneration of transformed plantlets was also achieved for all four cultivars although the frequency was much higher for Golden Promise than for the other three genotypes, reiterating that genotype is an important determinant in the regenerative ability of barley. This study has demonstrated for the first time that Agrobacterium-mediated transformation can be used to transform the Australian cultivars Sloop and Chebec. PMID- 14530865 TI - Rescue of shoot apical meristems of chrysanthemum by culturing on root tips. AB - A new method to regenerate plants from leaf primordia-free shoot apical meristem domes (LP-free SAMs) was developed by establishing the meristem dome on the cut surface of root tips. Ten days after culture, the viable rate of LP-free SAMs of chrysanthemum 'Piato' attached to chrysanthemum root tips was >40%. Shoot regeneration was not observed from LP-free SAMs without the root tips. When LP free SAMs of chrysanthemum were transferred to root tips of either petunia, cabbage, or carnation, the highest shoot regeneration rate was observed with cabbage root tips. Microscopic observation documented that the LP-free SAM temporarily adhered to the cut surface of the root tip of cabbage. PMID- 14530866 TI - Increased sucrose permeability in Behcet's disease. AB - Behcet's disease (BD) is a chronic, relapsing, systemic vasculitis with unknown etiology. During the progression of the disease, gastrointestinal involvement can be observed. The aim of this study was to find out the predictive value of the sucrose permeability test in detecting gastrointestinal mucosal damage in BD. Twenty-six male Behcet's patients and 21 age- and sex-matched controls were enrolled in the study. Seventeen patients had active disease, while nine did not. Active disease was defined as having elevated erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) levels or at least two of the diagnostic criteria in the previous month. Patients and controls were investigated with the upper gastrointestinal permeability test. Of all the patients, 25 were investigated endoscopically. The urine sucrose levels were significantly higher in patients than in the control group (P = 0.0001) and in patients with active disease than those with inactive disease (P < 0.0001), while urine sucrose of patients with inactive disease and the control group did not differ. The endoscopic findings were not specific to BD. Active and inactive BD had similar endoscopic findings. Increased upper gastrointestinal permeability was established in patients with BD. This increased permeability was not related to a specific gastrointestinal BD lesion. Further studies with larger series must be performed in order to determine the value of the sucrose permeability test in detecting mucosal involvement in BD. PMID- 14530867 TI - An open, randomized comparison study of cyclosporine A, cyclosporine A + methotrexate and cyclosporine A + hydroxychloroquine in the treatment of early severe rheumatoid arthritis. AB - PURPOSE: To determine whether a regimen of cyclosporine (CSA) and methotrexate (MTX), or CSA and hydroxychloroquine (HCQ) introduced in early rheumatoid arthritis (RA) can produce a significant improvement in clinical outcome and/or retard radiographic damage in comparison with standard monotherapy with CSA alone. METHODS: One hundred five patients with active RA of less than 36 months duration, who had never previously been treated with immunosuppressive agents, were included in a 12-month, multi-center, open, randomized trial. Patients who fulfilled the criteria for early severe RA were randomized to receive either combination therapy (CSA + MTX n = 34, CSA + HCQ n = 35) or CSA alone (n = 36). RESULTS: CSA + MTX was more effective than the other two treatment groups in controlling RA symptoms. CSA+MTX did not show a significant radiographic progression according to Larsen-Dale (0.90 +/- 3.89 compared to baseline values, P > 0.05); moreover, patients treated with CSA alone or CSA+HCQ showed a significant worsening of Larsen-Dale score (2.91 +/- 5.99 and 2.97 +/- 4.28 respectively vs baseline values, P < 0.05), although not significant when compared with the CSA + HCQ group (P = 0.56 and 0.39, respectively). CONCLUSIONS: This trial indicated that CSA+MTX was more effective than the other two treatments in improving clinical data and inhibiting radiographic progression, although the differences were not significant in this relatively small study. However, the difference was significant in favor of CSA + MTX regarding ACR 50% response. PMID- 14530868 TI - Cloning and biochemical characterization of hexokinase from the methylotrophic yeast Hansenula polymorpha. AB - We previously showed that, unlike other yeasts, Hansenula polymorpha possesses a glucokinase HPGLK1 that can mediate glucose repression in this yeast, although it cannot replace the regulatory function of hexokinase 2 in Saccharomyces cerevisiae. In the present study, the H. polymorpha hexokinase gene HPHXK1 was cloned by complementation of the glucose growth deficiency of the H. polymorpha double kinase-negative mutant A31-10 with a genomic library. The sequence of the 483-amino acid hexokinase protein deduced from the HPHXK1 gene showed the highest degree of identity (56%) with hexokinase from Schwanniomyces occidentalis, whereas the identity with hexokinase from Kluyveromyces lactis and both hexokinases from Sac. cerevisiae was 55%. The hexokinase protein was purified from crude extracts of H. polymorpha, using ion exchange chromatography and gel filtration. The K(m) values of the purified enzyme for glucose, fructose and ATP were 0.26 mM, 1.1 mM and 0.32 mM, respectively. H. polymorpha hexokinase was inhibited by trehalose-6-phosphate ( K(i)=12 microM) and ADP ( K(i)=1.6 mM), but not by glucose-6-phosphate. Transformation of a H. polymorpha hexokinase-negative mutant with a plasmid carrying the HPHXK1 gene restored the ability of the mutant to phosphorylate fructose and to repress the synthesis of alcohol oxidase and catalase by fructose. Therefore, hexokinase is specifically needed for the establishment of fructose repression in H. polymorpha. PMID- 14530869 TI - P-glycoprotein expression does not change the apoptotic pathway induced by curcumin in HL-60 cells. AB - PURPOSE: One of the mechanisms responsible for the multidrug resistance (MDR) phenotype of cancer cells is overexpression of so-called ATP-dependent drug efflux proteins: the 170-kDa P-glycoprotein (P-gp) encoded by the MDR1 gene and the 190-kDa multidrug resistance-associated protein 1 encoded by the MRP1 gene. The purpose of the present study was to verify the hypothesis postulating that P gp expression, apart from enabling drug efflux, confers on the cells resistance to apoptosis by inhibiting caspase-8 and caspase-3. MATERIALS AND METHODS: Human HL-60 cells, either drug-sensitive or with the MDR phenotype caused by overexpression of P-gp (HL-60/Vinc) or MRP1 (HL-60/Adr), were treated with the natural dye curcumin at 50 micro M or with UVC to induce apoptosis. Symptoms of cell death were assessed by morphological observation after Hoechst staining, DNA fragmentation was measured by flow cytometry and the TUNEL method, and caspase-8 and caspase-3 activation and cytochrome c release from mitochondria were measured by Western blotting. RESULTS: Curcumin induced cell death in HL-60 cells, both sensitive and with the MDR phenotype, which could be classified as caspase-3 dependent apoptosis, together with cytochrome c release, activation of caspase-3 and oligonucleosomal DNA fragmentation. No active caspase-8 was detected. Also UVC caused caspase-3 activation in both the sensitive and the MDR HL-60 cells. CONCLUSIONS: Our findings show that there was no correlation between P-gp expression and resistance to caspase-3-dependent apoptosis induced by curcumin and UVC, at least in HL-60 cells. However, we cannot exclude the possibility of parallel P-gp expression and caspase-3 inhibition in some other cell lines, as cancer cells can acquire many different apoptosis-resistance mechanisms. PMID- 14530870 TI - Citrate enhances the protective effect of orally administered bismuth subnitrate against the nephrotoxicity of cis-diamminedichloroplatinum. AB - Attenuation of the renal toxicity of cis-diamminedichloroplatinum (CDDP) is important in the use of this effective but cytotoxic anticancer agent. We have previously shown that the renal toxicity of CDDP can be efficiently reduced by the induction of metallothionein (MT) by preadministration of bismuth compounds in mice. Bismuth subnitrate (BSN) is used as an antigastric ulcer agent and as an antidiarrheic agent, and is suitable for inducing MT in the kidney in cancer patients. However, due to the low absorption rate of Bi from the gastrointestinal tract, the efficacy of BSN in inducing renal MT is low. In the present study, we examined the effects of citrate as a vehicle for oral administration of BSN on the tissue distribution of Bi and induction of MT in the kidneys and tumors in mice inoculated with Meth-A fibrosarcoma. Renal levels of MT and Bi were markedly increased in the mice given BSN dissolved in citrate solution compared with those given BSN suspended in saline. On the other hand, the use of citrate increased Bi accumulation in the tumor only slightly and did not increase tumor MT levels. Administration of BSN with citrate efficiently depressed the renal toxicity of CDDP, but did not affect its antitumor activity. Since both BSN and citrate are used clinically as pharmaceuticals, the combination regimen of BSN and citrate may be readily applicable as a countermeasure against the adverse side effects of CDDP without affecting its antitumor activity. PMID- 14530871 TI - Thrombopoietin stimulates ex vivo expansion of mature neutrophils in the early stages of differentiation. AB - We examined the effects of thrombopoietin (TPO) in combination with stem cell factor (SCF), interleukin-3 (IL-3), and granulocyte colony-stimulating factor (G CSF) on the proliferation and differentiation of human neutrophils. Purified CD34(+) hematopoietic progenitor cells were cultivated with SCF, IL-3, and G-CSF for 7 days (early phase), and thereafter nonadherent cells were further cultivated for 9 days with G-CSF alone (late phase). A large number of highly selected neutrophils (>95%) was obtained on day 16. We compared the expansion capacity in the presence or absence of TPO in each culture phase. The significantly larger number of neutrophils was obtained in the presence of TPO in the early culture phase. The number of expanded cells plateaued at day 16. Ultimately, a 550-fold increase in the number of neutrophils was achieved. These neutrophils gained the ability to respond effectively with chemotaxis and superoxide release, and were appropriately primed by G-CSF, granulocyte macrophage colony-stimulating factor, tumor necrosis factor-alpha, and IL-1beta for enhanced release of O(2)(-). The responsiveness of these cells was identical to that of peripheral blood neutrophils. However, TPO did not accelerate the maturation of neutrophils supported by G-CSF in the late phase of culture. Furthermore, priming effects and triggering effects of TPO on the production of superoxide metabolites from peripheral blood neutrophils were not observed. These results suggest that TPO regulates the proliferation and differentiation of neutrophils in the early stages, but not the late stages, of differentiation. PMID- 14530872 TI - Prior and concurrent administration of recombinant human megakaryocyte growth and development factor in patients receiving consolidation chemotherapy for de novo acute myeloid leukemia--a randomized, placebo-controlled, double-blind safety and efficacy study. AB - Pegylated recombinant human megakaryocyte growth and development factor (PEG rHuMGDF) administered after acute myeloid leukemia (AML) chemotherapy (CT) failed to shorten the time of transfusion-dependent thrombocytopenia in a previous study. In this multicenter, randomized, placebo-controlled, double-blind study we determined the effect of administration of PEG-rHuMGDF prior to CT and of administration prior, concurrent, and 1 day post CT on platelet recovery and transfusion requirements in patients receiving consolidation CT for de novo AML. Patients were randomized to receive either 30 microk/kg PEG-rHuMGDF as a single dose on day -6 ( n=37), placebo as a single dose on day -6 ( n=9), 30 microk/kg PEG-rHuMGDF administered on day -6 followed by 10 microg/kg on days -5 to day 6 (through CT and including the day after CT, n=35), or placebo administered on day -6 to day 6 ( n=9). The median times to transfusion-independent platelet recovery to >20x10(9)/l were 24.5 and 24.0 days in the PEG-rHuMGDF day -6 group and PEG rHuMGDF day -6 to 6, respectively, compared to 21.0 days in the placebo group. There were no significant differences in the number of days of platelet transfusions between either PEG-rHuMGDF schedule or placebo. The PEG-rHuMGDF day 6 to 6 group had a delayed absolute neutrophil count (ANC) recovery compared to either placebo or PEG-rHuMGDF day -6 treated patients. Thus, alteration of the scheduling of PEG-rHuMGDF in terms of earlier dosing before and during chemotherapy did not improve platelet recovery but rather delayed hematopoietic reconstitution. Although unexpected, these observations may be of major relevance for the design of future clinical trials with recombinant thrombopoietins. PMID- 14530874 TI - Focal splenic lesions in myeloproliferative disease: association with fatal outcome. AB - BACKGROUND: A nodular tumor of the spleen in patients with myeloproliferative disease (MPD) is a very rare form of splenic involvement. The aim of the study was to describe the clinical data, sonographic patterns, and prognosis of nodular splenic infiltration in patients with MPD. MATERIALS AND METHODS: During a 20 year period, nodular splenic lesions were found in 10 out of 183 patients with MPD. Retrospectively, splenic size, echomorphology of the lesions, clinical data, sonographic follow-up, and survival were analyzed. RESULTS: In 9 out of 10 patients the lesions were hyperechoic--in one patient hypoechoic. In 3 patients the lesions were solitary. Seven patients had multiple nodular lesions. Low platelet count was seen in 8 patients; blast crisis was seen in 7 patients. The mean survival time was 2.9 months after detection of the splenic lesions. In one patient, autopsy confirmed the diagnosis of myelosarcoma of the spleen. CONCLUSION: The appearance of nodular splenic lesions in MPD is associated with blast crisis and a short survival. Definite histologic or cytologic findings associated with splenic nodules in MPD have not been identified yet. Myelosarcoma of the spleen is the most probable diagnosis suggested. PMID- 14530876 TI - Lytic infection of Epstein-Barr virus (EBV) in hemophagocytic syndrome associated with EBV-induced lymphoproliferative disorder. AB - This report describes lytic infection with Epstein-Barr virus (EBV) in three cases of hemophagocytic syndrome (HPS) presumably associated with EBV-induced lymphoproliferative disorder. All cases were previously healthy females with ages ranging from 52 to 87 years who showed a fulminant clinical course with accompanying fever, liver dysfunction, and disseminated intravascular coagulation. Cases 1 and 2 showed proliferation of atypical T lymphocytes, and case 3 showed proliferation of atypical B lymphocytes. Hemophagocytic histiocytes were observed in the bone marrow, lymph nodes, spleen, and liver. Atypical lymphocytes in all cases showed a positive reaction for both EBV-encoded small RNA (EBER) indicating latent infection with EBV and immediate early mRNAs of the Bam HI fragment of lower stranded frame (BHLF), indicating lytic infection by in situ hybridization. Interestingly, BHLF-positive cells were predominant in all cases. It is possible that reactivation of EBV infection may be involved in triggering the induction of cytokines and abnormal activation of histiocytes. PMID- 14530877 TI - Antierythropoietin antibodies in thalassemia patients. AB - We evaluated sera from 58 thalassemic patients for the presence of antierythropoietin antibodies to investigate whether these autoantibodies may relate with modest response to treatment with recombinant human erythropoietin (rhEpo). Thirty-seven patients had beta-thalassemia major, 9 patients had beta thalassemia intermedia, and 12 patients had sickle/beta(+)-thalassemia. Of 58 patients, 24 were on rhEpo treatment in order to increase the intervals between transfusions or the hemoglobin (Hb) values. The study population was divided into three groups according to rhEpo treatment. Group A consisted of 15 patients who were on rhEpo treatment (400-600 IU/kg three times per week, subcutaneously) showing an increase of Hb values or reduction of transfusion requirements. Group B included 9 patients who did not respond to rhEpo and group C consisted of 34 patients who did not receive rhEpo. Laboratory studies included a complete blood count, measurement of serum erythropoietin, immunological evaluation, and determination of antierythropoietin antibodies using enzyme-linked immunosorbent assay (ELISA). There were no significant differences among groups A, B, and C concerning age, Hb, and endogenous erythropoietin values. Fifteen patients had positive antinuclear antibodies and three patients had positive rheumatoid factor. Antierythropoietin antibodies were detected in the sera of seven patients (five men and two women) who received rhEpo. The male patients and one female patient had no response to rhEpo while the other female patient showed response when the dose increased. Other autoantibodies seem to have no clinical significance. In the present study, we detected for the first time in thalassemia patients the presence of antierythropoietin antibodies, which may contribute to rhEpo resistance. Thalassemia patients with low response rates to rhEpo should be evaluated for the presence of antierythropoietin antibodies. PMID- 14530878 TI - Hematopoietic stem cell transplantation for severe aplastic anemia--experience of an institute in Taiwan. AB - Hematopoietic stem cell transplantation (HSCT) is an accepted treatment strategy for patients with severe aplastic anemia (SAA). We report our experience in a general hospital in Taiwan. From March 1985 to July 2001, 79 consecutive SAA patients, 46 male and 33 female, with a median age of 22 (4-43) years, received 80 courses of transplantation. Cyclophosphamide and total body radiation were used for the conditioning regimen, and cyclosporine-A and methotrexate for graft versus-host disease (GVHD) prevention. Patients were followed for a median of 39 months (from 8 days to 194 months). Myeloid and platelet engraftment occurred in a median of 15 (8-27) days and 18 (8-77) days, respectively. Three patients had primary and three patients secondary graft failure. Five patients (6.8%) had grade II-IV acute GVHD in 73 evaluable patients. Chronic GVHD occurred in 23 (34.8%) patients, with extensive stage in six. Only two patients had CMV disease. The projected 3- and 5-year overall survival rates estimated by the Kaplan-Meier method were 76.08 and 74.13%, respectively. Age at transplant, non-sibling donor, mononuclear cell dose, grade II-IV acute GVHD, interval from diagnosis to transplant, and red blood cell and platelet transfusion before transplant were poor prognostic factors for overall survival by univariate analysis. Grade II-IV acute GVHD was the only prognostic factor affecting overall survival after multivariate Cox regression analysis (P=0.040). In conclusion, SAA patients receiving HSCT have good long-term survival. The low incidence of acute GVHD in our patients may be related to ethnicity. PMID- 14530879 TI - High-risk AML complicated by pulmonary aspergillosis: successful treatment with nonmyeloablative stem cell transplantation and long-term administration of voriconazole. AB - Acute myeloid leukemia (AML) associated with central diabetes insipidus (DI) and chromosomal aberrations is characterised by a very poor prognosis. We present a 28-year-old female with AML FAB M0, preceding DI and cytogenetic abnormalities (monosomy 7 and inversion of chromosome 9). Complete remission was achieved with FLAG after she was refractory to two different induction regimens. Prolonged neutropenia resulted in invasive pulmonary aspergillosis. Allogeneic stem cell transplantation from a matched unrelated donor was performed using a reduced intensity conditioning regimen. Desmopressin substitution for DI was withdrawn after transplant without recurrence of symptoms. Initial antifungal treatment, including liposomal amphotericin B, caspofungin and itraconazole, was replaced by voriconazole after deterioration of pulmonary aspergillosis, resulting in improvement, stabilisation and finally, also as the combined effect of discontinuation of the immunosuppressive therapy, in disappearance of signs and symptoms. Thirteen months after transplant, the patient is in continuous complete remission. The presented case study thus demonstrates that high-risk AML with concomitant invasive fungal infection may be safely and effectively treated by nonmyeloablative stem cell transplantation and long-term administration of voriconazole. PMID- 14530881 TI - Intertrochanteric osteotomy in young adults for sequelae of Legg-Calve-Perthes' disease--a long term follow-up. AB - Between 1974 and 1999 we performed 15 intertrochanteric osteotomies in 14 patients with a painful hip secondary to Legg-Calve-Perthes' disease. In seven patients, the osteotomy was combined with advancement of the greater trochanter, acetabular roof plasty, or both. One patient died 5 years after the osteotomy and one patient was lost to follow-up. One patient had a revision osteotomy 21 years after the initial osteotomy, and five patients had a prosthetic replacement 8-25 years after the osteotomy. The average follow-up of the remaining osteotomies was 11.3 years. An intertrochanteric osteotomy can decrease pain and improve clinical function medium and long term without radiological progression of joint degeneration. PMID- 14530882 TI - Clear cell chondrosarcoma: radiographic, computed tomographic, and magnetic resonance findings in 34 patients with pathologic correlation. AB - OBJECTIVE: To describe the radiographic features of clear cell chondrosarcoma (CCCS), including the computed tomographic (CT) and magnetic resonance (MR) findings, and to correlate them with the histopathologic findings. DESIGN AND PATIENTS: A retrospective review was carried out of 72 patients with histopathologically confirmed CCCS. Imaging studies were available for 34 patients: conventional radiographs (n=28), CT scans (n=14), and MR images (n=15). Radiographic studies were reviewed by three radiologists who rendered a consensus opinion; the studies were correlated with the histopathologic findings. RESULTS: Of the 34 patients with imaging studies, 30 were male and 4 were female (mean age 38.6 years; range 11-74 years). Twenty-two lesions were in long bones (15, proximal femur; 1, distal femur; 1, proximal tibia; 5, proximal humerus) and 11 were in flat bones (5, vertebra; 4, rib; 1, scapula; 1, innominate). One lesion occurred in the tarsal navicular bone. Typically, long bone lesions were located in the epimetaphysis (19/22) and were lucent with a well-defined sclerotic margin and no cortical destruction or periosteal new bone formation. More than one-third of the long bone lesions contained matrix mineralization with a characteristic chondroid appearance. Pathologic fractures were present in six long bone lesions (4, humerus; 2, femur). Lesions in the proximal humerus were more likely to have indistinct margins (4/5) and extend into the diaphysis. Flat bone lesions were typically lytic and expansile and occasionally demonstrated areas of cortical disruption. Typically, matrix mineralization, when present, was amorphous. MR imaging, when available, was superior to conventional radiographs for demonstrating the intramedullary extent of a lesion as well as soft tissue extension. CT images better delineated the presence of cortical destruction and the character of matrix mineralization patterns. CCCS lesions were typically low signal intensity on T1-weighted images and moderately or significantly bright on T2-weighted images. Areas of lesion heterogeneity on T1- and T2-weighted images and on post-gadolinium T1-weighted images corresponded pathologically to areas of mineralization, intralesional hemorrhage, and cystic changes. Adjacent bone marrow edema was typically absent (12/15) or only minimally observed in a few cases (3/15). No cases examined with MR imaging demonstrated periosteal new bone formation. CONCLUSIONS: CCCS typically presents radiographically as a geographic lytic lesion located in the epimetaphyseal region of long bones. Most commonly lesions are found in the proximal femur, followed by the proximal humerus. Lesions within the proximal humerus may exhibit more aggressive features. Lesions in the axial skeleton are typically expansile and destructive, often with soft tissue extension and lack of mineralization. MR imaging may show the presence or absence of bone marrow edema. PMID- 14530883 TI - Genomic analysis of immunity in a Urochordate and the emergence of the vertebrate immune system: "waiting for Godot". AB - Genome-wide sequence analysis in the invertebrate chordate, Ciona intestinalis, has provided a comprehensive picture of immune-related genes in an organism that occupies a key phylogenetic position in vertebrate evolution. The pivotal genes for adaptive immunity, such as the major histocompatibility complex (MHC) class I and II genes, T-cell receptors, or dimeric immunoglobulin molecules, have not been identified in the Ciona genome. Many genes involved in innate immunity have been identified, including complement components, Toll-like receptors, and the genes involved in intracellular signal transduction of immune responses, and show both expansion and unexpected diversity in comparison with the vertebrates. In addition, a number of genes were identified which predicted integral membrane proteins with extracellular C-type lectin or immunoglobulin domains and intracellular immunoreceptor tyrosine-based inhibitory motifs (ITIMs) and immunoreceptor tyrosine-based activation motifs (ITAMs) (plus their associated signal transduction molecules), suggesting that activating and inhibitory receptors have an MHC-independent function and an early evolutionary origin. A crucial component of vertebrate adaptive immunity is somatic diversification, and the recombination activating genes (RAG) and activation-induced cytidine deaminase (AID) genes responsible for the Generation of diversity are not present in Ciona. However, there are key V regions, the essential feature of an immunoglobulin superfamily VC1-like core, and possible proto-MHC regions scattered throughout the genome waiting for Godot. PMID- 14530884 TI - Evolution of the proto-MHC ancestral region: more evidence for the plesiomorphic organisation of human chromosome 9q34 region. AB - The present day structure of the vertebrate major histocompatibility complex (MHC) and its three paralogous regions has always been a focus of interest. In a recent study, nine human anchor genes located in the MHC region were cloned from a Branchiostoma floridae (amphioxus) cosmid library. The identification and analysis of 31 surrounding genes led to the most probable model of two rounds of en bloc duplication giving rise to these regions. These events were estimated to have occurred after the cephalochordata-craniata divergence [approximately 766 million years ago (Mya)] and before the Gnathostomata radiation (approximately 528 Mya). Furthermore, it was also shown that after this large-scale duplication one of these regions, corresponding to the human 9q33-q34, had retained an ancestral organisation. In the present study, four new cosmids in the amphioxus proto-MHC region were identified by the chromosomal walking technique. These cosmids were sequenced, and their structural annotation was performed, leading to the prediction of eleven genes. Their phylogenetic relationships among species corroborate the results obtained previously and provide more evidence for the plesiomorphic state of the human chromosome 9q33-34 MHC paralogous region. PMID- 14530886 TI - Nephrogenic rests mimicking Wilms' tumor on CT. AB - Nephrogenic rests (NR) are persistent benign remnants of embryonic renal tissue. A small percentage of these may develop into Wilms' tumor (WT). Radiologic imaging is relied upon to differentiate between these entities, with the hallmark of malignant transformation being growth on serial imaging studies. There is, however, considerable overlap in their imaging characteristics. The authors present a case of two biopsy-proven NR in a 2-year-old girl with sporadic aniridia that were indistinguishable from WT on initial radiologic studies. One of the NR grew on serial imaging studies mimicking a WT, but after resection was confirmed to be a benign hyperplastic NR on pathologic examination. PMID- 14530885 TI - Frequent segmental sequence exchanges and rapid gene duplication characterize the MHC class I genes in lemurs. AB - Major histocompatibility complex (MHC) class I genes have complicated and profound evolutionary histories. To reconstruct and better understand their histories, partial class I genes (exon 2-intron 2-exon 3) were sequenced in a sampling of prosimians (Strepsirhini, Primates). In total, we detected 117 different sequences from 36 Malagasy prosimians (lemurs) and 1 non-Malagasy prosimian (galago) representing 4 families, 7 genera, and 13 species. Unlike the MHC class II genes ( MHC-DRB), MHC class I genes show a generally genus-specific mode of evolution in lemurs. Additionally, no prosimian class I loci were found to be orthologous to HLA genes, even at highly conserved loci (such as HLA-E, HLA F). Phylogenetic analysis indicates that nucleotide diversity among loci was very small and the persistence time of the polymorphisms was short, suggesting that the origin of the lemur MHC class I genes detected in this study was relatively recent. The evolutionary mode of these genes is similar to that of classic HLA genes, HLA-A, HLA-B, and HLA-C, in terms of their recent origin and rarity of pseudogenes, and differs from them with respect to the degree of gene duplications. From the viewpoint of MHC genes evolution, some interlocus sequence exchanges were apparently observed in the lemur lineage upon phylogenetic and amino acid motif analyses. This is also in contrast to the evolutionary mode of HLA genes, where intralocus exchanges have certainly occurred but few interlocus exchanges have taken place. Consequently, the gene conversion model for explaining the generation of the MHC diversity among different loci can be thought to play more important roles in the evolution of lemur MHC class I genes than in that of HLA genes. PMID- 14530887 TI - MR imaging features of craniodiaphyseal dysplasia. AB - We report the magnetic resonance (MR) imaging findings in a 4-year-old girl with characteristic radiographic and computed tomography (CT) features of craniodiaphyseal dysplasia. MR imaging exquisitely depicted cranial nerve compression, small foramen magnum, hydrocephalus, and other intracranial complications of this syndrome. A syrinx of the cervical spinal cord was demonstrated. We suggest that MR imaging become a routine component of the evaluation of these patients. PMID- 14530888 TI - Ultrasound-guided percutaneous catheter drainage of splenic abscess. AB - Splenic abscess is an uncommon clinical condition which is usually treated with splenectomy with its attendant risks. Image-guided percutaneous drainage of splenic abscesses has been described in adults. The case of a 12-year-old boy with a splenic abscess who was successfully treated with ultrasound-guided percutaneous catheter drainage is presented. Image-guided percutaneous drainage in children can be a safe and effective alternative to surgery. PMID- 14530889 TI - Age dependence of the renal apparent diffusion coefficient in children. AB - BACKGROUND: Diffusion imaging has proved to be a powerful tool for diagnosing ischemic lesions in the brain, and the technique is now being applied to other organs, including the kidneys. For quantitative studies it is important to define the normal values of the apparent diffusion coefficient (ADC), given the important physiological changes that occur in the kidney during early childhood it is likely that the ADC changes markedly during this period. OBJECTIVE. To evaluate the age dependent changes in the ADC of normal kidneys in the pediatric population. MATERIALS AND METHODS: The whole kidney ADC was calculated for 62 pediatric patients on a 1.5-T system using a respiratory-triggered, single-shot diffusion tensor imaging sequence with b values of 50, 200, and 350 mm(2)/s. RESULTS. The ADC was found to increase with age with the largest increase being in the first year of life, the rate of change being described by a constant plus a power function, specifically 1349+[358.5*[age 0.34]], ( P<0.001). CONCLUSION: The renal ADC changes significantly during childhood. PMID- 14530892 TI - Pharmacokinetics and metabolism of 14C-levetiracetam, a new antiepileptic agent, in healthy volunteers. AB - The absorption, disposition and metabolism of levetiracetam, a new antiepileptic drug, have been investigated after a single oral dose of the (14)C-labelled molecule administered to male healthy volunteers. As chiral inversion can occur during drug metabolism, the chiral inversion of levetiracetam and/or of its major metabolite produced by hydrolysis (the corresponding acid) was also investigated. Finally, the in vitro hydrolysis of levetiracetam to its major metabolite and the inhibition of this reaction in human blood have been studied. Levetiracetam was very rapidly absorbed in man, with the peak plasma concentration of the unchanged drug occurring at 0.25-0.50 h. The unchanged drug accounted for a very high percentage of plasma radioactivity (97-82%) at all the times measured, i.e. until 48 h after administration. The apparent volume of distribution of the compound was close (0.55-0.62 l/kg) to the volume of total body water. Total body clearance (0.80-0.97 ml/min/kg) was much lower than the nominal hepatic blood flow. The plasma elimination half-life of the unchanged drug varied between 7.4 h and 7.9 h. Plasma to blood ratio of total radioactivity concentrations was 1.1 1.3, showing that radioactivity concentrations were similar in blood cells and plasma. The balance of excretion was very high in all four volunteers. The predominant route of excretion was via urine, accounting for a mean of 95% of the administered dose after 4 days. Two major radioactive components were present in urine, the unchanged drug and the acid obtained by hydrolysis, accounting for 66% and 24% of the dose after 48 h, respectively. Hydrolysis of levetiracetam in human blood followed Michaelis-Menten kinetics with Km and V(max) values of 435 microM and 129 pmol/min/ml blood, respectively. Among the inhibitory agents investigated in this study, only paraoxon inhibited levetiracetam hydrolysis (92% inhibition at 100 microM). Oxidative metabolism occurred in man, although it accounted for no more than 2.5% of the dose. There was no evidence of chiral inversion. PMID- 14530893 TI - Adaptation of reach-to-grasp movement in response to force perturbations. AB - This study examined how reach-to-grasp movements are modified during adaptation to external force perturbations applied on the arm during reach. Specifically, we examined whether the organization of these movements was dependent upon the condition under which the perturbation was applied. In response to an auditory signal, all subjects were asked to reach for a vertical dowel, grasp it between the index finger and thumb, and lift it a short distance off the table. The subjects were instructed to do the task as fast as possible. The perturbation was an elastic load acting on the wrist at an angle of 105 deg lateral to the reaching direction. The condition was modified by changing the predictability with which the perturbation was applied in a given trial. After recording unperturbed control trials, perturbations were applied first on successive trials (predictable perturbations) and then were applied randomly (unpredictable perturbations). In the early predictable perturbation trials, reach path length became longer and reaching duration increased. As more predictable perturbations were applied, the reach path length gradually decreased and became similar to that of control trials. Reaching duration also decreased gradually as the subjects adapted by exerting force against the perturbation. In addition, the amplitude of peak grip aperture during arm transport initially increased in response to repeated perturbations. During the course of learning, it reached its maximum and thereafter slightly decreased. However, it did not return to the normal level. The subjects also adapted to the unpredictable perturbations through changes in both arm transport and grasping components, indicating that they can compensate even when the occurrence of the perturbation cannot be predicted during the inter-trial interval. Throughout random perturbation trials, large grip aperture values were observed, suggesting that a conservative aperture level is set regardless of whether the reaching arm is perturbed or not. In addition, the results of the predictable perturbations showed that the time from movement onset to the onset of grip aperture closure changed as adaptation occurred. However, the spatial location where the onset of finger closure occurred showed minimum changes with perturbation. These data suggest that the onset of finger closure is dependent upon distance to target rather than the temporal relationship of the grasp relative to the transport phase of the movement. PMID- 14530894 TI - Inharmonicity detection. Effects of age and contralateral distractor sounds. AB - Detection of mistuned partials in otherwise harmonic complex tones was investigated in naive subjects of three different age groups. Signals were presented at constant sensation level to compensate for differences in hearing sensitivity and to specifically examine age-related changes in inharmonicity perception. Performance was measured under two conditions, monaural signal presentation and dichotic signal-noise presentation, with the latter aiming at the influence of contralateral distractor sounds. Stimuli were complex tones with ten harmonics and 125-Hz fundamental frequency. Mistuning detection was measured for the first, second, fourth, and eighth harmonic. In a three-interval, three alternative forced-choice procedure, subjects were required to distinguish a complex tone containing one mistuned partial from two reference tones, with all partials at their harmonic frequencies. Thresholds were measured as the amount of frequency shift necessary for the mistuning to be detected. Performance deteriorated moderately with age for the two higher partials tested, but not for the lower ones. Thresholds for dichotic signal/noise presentation did not differ significantly from monaural ones in any of the age groups. Results are discussed in relation to hypotheses of harmonicity perception in auditory scene analysis and with respect to the investigation of patients suffering form respective deficits due to acquired brain lesions. PMID- 14530895 TI - Do vertical shifts in dose-response rate-relationships in operant conditioning procedures indicate "sensitization" to "drug wanting"? PMID- 14530901 TI - Acute effects of memantine in combination with alcohol in moderate drinkers. AB - RATIONALE: Alcohol effects in humans involve N-methyl- d-aspartate (NMDA) receptor-mediated glutamatergic neurotransmission. It has been proposed that NMDA receptor antagonists may be effective in the treatment of alcohol dependence. OBJECTIVE: This study evaluated the acute effects of memantine, an NMDA receptor antagonist, on the subjective, physiological, and performance effects of alcohol in moderate (10-30 drinks per week) alcohol drinkers. METHODS: Eighteen volunteers without alcohol dependence were tested using a double-blind design with three 3-day long inpatient phases separated by at least a 2-week wash-out period. Memantine (0, 15, and 30 mg) was administered 4 h before alcohol (1.5 g/l body water), which was given in four divided doses every 20 min. RESULTS: Pretreatment with memantine attenuated the craving for alcohol before alcohol administration, but not after alcohol was given. Memantine increased the dissociative effects of alcohol, without altering its sedative, stimulant, and overall intoxicating effects. Memantine also did not affect alcohol-induced impairment in performance, physiological changes, or pharmacokinetics. Memantine increased subjective reports of dissociation, confusion, and stimulation, and impaired motor coordination on the balance task. CONCLUSIONS: Memantine was well tolerated in combination with alcohol. The findings suggest that NMDA receptor neurotransmission may be involved in alcohol craving and alcohol-induced subjective dissociative effects. PMID- 14530902 TI - Nicotine as a signal for the presence or absence of sucrose reward: a Pavlovian drug appetitive conditioning preparation in rats. AB - RATIONALE: In Pavlovian conditioning research, nicotine is typically conceptualized as the unconditioned stimulus (US) that becomes associated with an exteroceptive conditioned stimulus (CS). This research has not explored the possibility that nicotine can also function as a CS. OBJECTIVES: The present research examined whether nicotine served as a CS for the presence (CS+) or absence (CS-) of sucrose and started defining its specificity. METHODS AND RESULTS: Rats trained in the CS+ condition had nicotine (0.4 mg/kg, base) paired intermittently with brief access to sucrose. Intermixed were saline sessions without sucrose. Nicotine acquired the ability to evoke goal tracking. This conditioned response (CR) decreased across extinction sessions. The CR was sensitive to nicotine dose (ED(50)=0.113 mg/kg) and administration to testing interval; 0-min and 100-min delays produced no CR. The CS properties were specific to nicotine in that amphetamine and bupropion substitution was incomplete. Rats in the CS- condition received similar discrimination training except that sucrose was paired with saline. Nicotine also served as a CS-; the saline state CS+ acquired control of goal tracking. Mecamylamine, but not hexamethonium, blocked nicotine's ability to serve as a CS+ and CS-, indicating a role for central nicotinic acetylcholine receptors. CONCLUSIONS: Nicotine served as a signal for the presence or absence of sucrose. The extinction, CS-, and substitution results eliminated a psychomotor stimulant account. The conceptualization of nicotine as a CS suggests novel empirical research in which a drug acquires additional inhibitory and/or excitatory value based on other outcomes present during its effects. PMID- 14530904 TI - Modulation of Fas receptor proteins and dynamin during opiate addiction and induction of opiate withdrawal in rat brain. AB - The Fas receptor is involved in the regulation of apoptosis but also can function as a non-apoptotic signal transducer. This study was mainly designed to quantitate Fas proteins in rat brain during heroin addiction and opiate withdrawal. In rat, mouse and human brains, and in SH-SY5Y cells, similar forms of Fas were immunodetected with different antibodies (i.e., 35 kDa native Fas and 48- and 51-kDa glycosylated Fas). Acute (2 h) treatments with the micro-opioid receptor agonists heroin (10 mg/kg) and morphine (30 mg/kg) increased the immunodensity of native Fas (124% and 36%) but not that of glycosylated Fas in the cerebral cortex. Chronic (5 days) heroin (5-30 mg/kg) and morphine (10-100 mg/kg) were also associated with increased native Fas (76% and 45%) and with different expressions of glycosylated Fas. In heroin-dependent rats, opiate withdrawal (48 h) resulted in a sustained increase in native Fas (107%) and in up regulation of 51 kDa glycosylated Fas (51%). Acute treatments with selective delta-receptor (SNC-80, 10 mg/kg) or kappa-receptor (U 50488-H, 10 mg/kg) agonists did not alter the content of native or glycosylated Fas. Chronic pentazocine (10-80 mg/kg, 5 days), a mixed opiate drug and sigma(1) receptor agonist, decreased native (48%) and glycosylated (38-82%) Fas proteins. Similarly, the selective sigma(1) agonist (+)-SKF 10047 also decreased native Fas (37%) and the effect was blocked by the sigma(1) antagonist BD 1063. Brain dynamin was up-regulated by acute and/or chronic heroin (30-39%), morphine (47 85%), pentazocine (51%) and heroin withdrawal (74%). The main results indicate that chronic heroin/morphine treatment and heroin withdrawal are associated with up-regulation of 35 kDa native Fas (and with different expressions of glycosylated Fas), and also with concomitant increases of dynamin in rat brain. PMID- 14530903 TI - Enhancement of latent inhibition by two 5-HT2A receptor antagonists only when given at both pre-exposure and conditioning. AB - RATIONALE: Clozapine-like atypical antipsychotic drugs, such as olanzapine, risperidone and sertindole, bind most strongly to 5-HT(2A) receptors, which may contribute to their antipsychotic effects. Antipsychotic drugs, such as clozapine and haloperidol, have been found to enhance latent inhibition (LI) in humans and rats. LI is a process of learning to ignore irrelevant stimuli that is disrupted in acute, positive-symptom schizophrenia, and can be modelled in animals. OBJECTIVES: The aim of this study was to determine the effects of two selective 5 HT(2A) receptor antagonists, SR 46,349B and ICI 169,369, on LI, as a test of their antipsychotic potential. METHODS: Doses of the 5-HT(2A) receptor antagonists that were sufficient for receptor blockade were determined in 5-HT behavioural syndrome tests. SR 46,349B and ICI 169,369 were then tested for enhancement of LI and reversal of amphetamine-induced attenuation of LI in a conditioned suppression paradigm. RESULTS: SR 46,349B (0.6-2.4 mg kg(-1) i.p.) and ICI 169,369 (10-40 mg kg(-1) i.p.) antagonised 5-hydroxytryptophan (5-HTP) induced head twitches and wet dog shakes, which are mediated by 5-HT(2A) receptors, but had no effect on mCPP-induced hypolocomotion, which is mediated by 5-HT(2C) receptors. Neither SR 46,349B (1.2 mg kg(-1) i.p.) nor ICI 169,369 (40 mg kg(-1) i.p) affected 8-hydroxy-2-(di- n-propylamino)tetralin (8-OH-DPAT) induced forepaw treading, suggesting that they were not in vivo 5-HT(1A) receptor antagonists. SR 46,349B (2.4 mg kg(-1) i.p.) and ICI 169,369 (40 mg kg(-1) i.p.) enhanced LI when given at both the pre-exposure and conditioning stages of the paradigm, but not when given at either pre-exposure or conditioning only. Both drugs also reversed the disruption of LI induced by D-amphetamine (1 mg kg(-1) i.p.). CONCLUSIONS: The profile of SR 46,349B and ICI 169,369 in LI differs from that of clozapine and haloperidol in LI, which both enhance LI when given only at the conditioning stage of the paradigm. PMID- 14530905 TI - Gene polymorphisms within the immune system that may underlie drug allergy. AB - Drug allergy encompasses a broad spectrum of different diseases. It occurs in some individuals, whereas it does not occur in many others. This suggests, among others, the involvement of hereditary factors, and thus of gene polymorphisms. Since drug metabolising enzymes as well as the immune system itself may be responsible for drug allergy, gene polymorphisms are relevant in both systems. While already some information exists on gene polymorphisms of drug metabolising enzymes that result in drug allergy, little information is available on gene polymorphisms within the immune system that result in such allergy. This review sets out to provide an avenue for future research aimed at discovering such polymorphisms. To this end, immune mechanisms that underlie drug allergy will be discussed. A pivotal mechanism underlying several types of drug allergy, immediate-type (type I) hypersensitivity, is also a hallmark of asthma, and therefore drug allergy and asthma share a range of candidate genes. Research on asthma has come relatively far in establishing associations of disease with polymorphisms in these genes. Therefore, these polymorphisms and their associations with asthma will be discussed. These studies on asthma provide us with lessons on how to conduct such studies on drug allergy. PMID- 14530906 TI - Evidence for a significant role of a Gs-triggered mechanism unrelated to the activation of adenylyl cyclase in the cyclic AMP-independent relaxant response of guinea-pig tracheal smooth muscle. AB - Cyclic AMP is a key molecule in the regulation of airway smooth muscle tone. Increased cyclic AMP leads to relaxation of this smooth muscle and its inhibition results in the muscle contraction. A constitutive role for cyclic AMP in the contraction and relaxation of airway muscle is supported by the observations that direct activators of adenylyl cyclase, such as forskolin and membrane-permeable cyclic AMP analogues, relax this smooth muscle potently. This traditional view of the role for cyclic AMP is the basis for the idea that relaxation of airway smooth muscle mediated through adenylyl cyclase-linked, G(s)-coupled receptors, including the beta(2)-adrenoceptor, is achieved mainly by the elevation of cyclic AMP content [cyclic AMP-dependent mechanism(s)]. However, recent pharmacological and biochemical evidence raises a fundamental question concerning the role of cyclic AMP; can G(s)-coupled receptor-mediated relaxation of tracheal smooth muscle be attributed exclusively to cyclic AMP-dependent mechanism(s)? In the present study, we show that cholera toxin (CTX, 5 microg/ml), an activator of the heterotrimeric guanine-nucleotide-binding protein G(s), relaxes guinea-pig tracheal smooth muscle. CTX also elevates tissue cyclic AMP content by about 30 fold and this is practically abolished by an adenylyl cyclase inhibitor, SQ 22,536 (100 microM). However, unexpectedly, the relaxant response to CTX is not affected by SQ 22,536. These results firstly show that activation of G(s) is able to produce a relaxation in tracheal smooth muscle independently of the elevation of cyclic AMP. G(s)-triggered, cyclic AMP-unrelated cellular mechanism(s) seem(s) to play a substantial role in smooth muscle relaxation mediated through adenylyl cyclase-linked receptors. This mechanism may account in part for the cyclic AMP independent relaxant response of tracheal smooth muscle. PMID- 14530907 TI - Characterization of the transport of the bicyclic peptide phalloidin by human hepatic transport proteins. AB - Phalloidin, the major phallotoxin of the mushroom Amanita phalloides, enters hepatocytes by a carrier-mediated mechanism. The molecular identity of the transport proteins mediating phalloidin uptake was so far unknown. Earlier studies in rat liver indicated that phalloidin may share a common mechanism of uptake with organic anions like bile salts. In the current study on human transporters, we analyzed the uptake of phalloidin into transfected HEK293 cells stably expressing the recombinant hepatocyte-specific organic anion uptake transporters OATP2 (also termed OATP1B1, OATP-C, LST1, symbol SLC21A6) or OATP8 (OATP1B3 or SLC21A8). Time-dependent uptake of phalloidin was observed with SLC21A6-expressing cells and was inhibited by typical substrates of SLC21A6 such as bromosulfophthalein or cholyltaurine. A K(m) value of 39+/-11 micro M was determined for SLC21A6-mediated phalloidin uptake. Additional inhibitors of phalloidin uptake mediated by SLC21A6 included the immunosuppressive drugs cyclosporin A, FK506, and rapamycin, whereas alpha-amanitin was only a weak inhibitor. Cyclosporin A was a most potent competitive inhibitor for SLC21A6 mediated phalloidin transport with a K(i) value of 51 nM. PMID- 14530908 TI - Prevalence of osteoporosis and its reproductive risk factors among Jordanian women: a cross-sectional study. AB - Extensive differences in the osteoporosis epidemiological pattern among geographic and ethnic groups have been reported. The evidence concerning association of multiple pregnancies, lactations, and other menstrual history factors with low bone mineral density (BMD) remains inconclusive. Previous local studies addressing these issues in Jordan are very restricted. We present a cross sectional study of Jordanian women who visited outpatient clinics between August 2000 and August 2002 at two community hospitals in Amman City. BMD measurement was performed for all subjects, while comprehensive appraisal of clinical issues related to reproductive status and past medical history was carried out using a structured questionnaire administered to 50% of the subjects. We also attempted to examine the current hypothesis of possible influence of hyperlipidemia and thyroid abnormalities on decreased BMD. According to WHO criteria, 119 (29.6%) were identified as having osteoporosis, 176 (43.8%) were osteopenic, and 107 (26.6%) had normal BMD. The multiple-linear regression analyses at different bone sites revealed that age, years of menopause, low-density lipoprotein (LDL), and follicle-stimulating hormone (FSH) have strong independent associations with decreased BMD at all lumbar and femoral neck regions. The negative effect associated with number of children (live births) and frequency of lactations was only evident at femoral neck. Although years of menstruation, age at menopause, days of menstrual cycle, number of pregnancies, and duration of hormone replacement therapy (HRT) were positively correlated with BMD, they had weaker associations than previous variables. Moreover, in the final multivariable logistic regression model, variables which rendered significantly independent risk factors after adjustment for age and BMI were: current smokers of more that 25 cigarettes/day, postmenopausal women irrespective of HRT use, menopausal years of > or =5 year intervals, natural early menopause, gastrointestinal disease, rheumatoid arthritis, osteoarthritis, hypertension, and thyroid replacement therapy. Ever-lactation, frequent lactation of 4 or more times, duration of lactation interval of 1-6 months and clinical hyperthyroidism were significant protective factors. Hysterectomy with or without oophorectomy, premature ovarian failure, gravidity, menstrual flow pattern, family history of osteoporosis, clinical hypothyroidism, hyperlipidemia, HRT, and corticosteroids therapy were not independent predictors of osteoporosis among our population. It was concluded that the prevalence of this worldwide public health problem among the Jordanian female population is extremely high, and is even found in younger age categories compared to previous international surveys. Though, the number of pregnancies in our multiparous female population showed a negative impact on femoral neck BMD, no evidence of increased risk of osteoporosis among ever-pregnant women was noted. Conversely, the current data analysis highlight many potential risk factors including associated medical illnesses, and other hormonal alterations experienced during menopausal period. Therefore, increased health awareness and intensive screening programs are mandatory for early detection of low bone mass. PMID- 14530909 TI - Peptic ulcer disease and calcium intake as risk factors of osteoporosis in women. AB - INTRODUCTION: Low dietary intake and decreased absorption of calcium are known as important risk factors of osteoporosis. Peptic ulcer disease may be accompanied by dietary restrictions influencing negatively calcium intake. Inflammation of gastric and duodenal mucosa as well as alkali used may significantly decrease calcium absorption. Additionally, bone metabolism may be changed by inflammatory mediators released as a result of mucosal inflammation. AIMS: Comparison of bone mineral density and calcium dietary intake in women with and without (control group) peptic ulcer disease. METHODS: Two hundred and sixty-three women were studied: 143 (mean age 60.3 years) with peptic ulcer disease diagnosed by endoscopy and/or upper gastrointestinal X-ray, and 120 (mean age 58.4 years) as controls. History of alimentary tract diseases and presence of risk factors of osteoporosis, as well as history of hormone replacement therapy, were collected based on specially designed questionnaires. Women with present risk factors of secondary osteoporosis and with previously diagnosed osteoporosis were excluded. The calcium dietary intake was determined using a standard questionnaire assessing milk and milk products intake as well as calcium supplementation when used. Bone mineral density of the lumbar spine and femoral bone was determined by DXA. RESULTS: Women with peptic ulcer disease not using hormone replacement therapy had lower bone mineral density in all studied regions as compared to control group without peptic ulcer disease. In the subgroup not using hormone replacement therapy all studied values differed significantly. In the smaller subgroup of women using hormone replacement therapy not all values were statistically significant. There was no statistical significance between studied groups in dietary calcium intake as milk, milk products, and calcium supplements. CONCLUSIONS: Calcium intake in women with ulcer disease is similar to healthy subjects. Peptic ulcer disease is an independent risk factor for osteoporosis in women. PMID- 14530910 TI - The contribution of hip fracture to risk of subsequent fractures: data from two longitudinal studies. AB - BACKGROUND: The contribution of hip fracture to the risk of subsequent fractures is unclear. METHODS: Data from the Baltimore Hip Studies and the Established Populations for Epidemiologic Studies of the Elderly (EPESE) were used. Baltimore subjects enrolled at the time of hip fracture ( n=549) and EPESE subjects without previous fractures at baseline ( n=10,680) were followed for 2-10 years. Self reported nonhip skeletal fracture was the outcome, and hip fracture was a time varying covariate in a survival analysis stratified by study site. The model was adjusted for race, sex, age, BMI, stroke, cancer, difficulty walking across a room, dependence in grooming, dependence in transferring, and cognitive impairment. RESULTS: The rate of all subsequent self-reported fractures after hip fracture was 10.4 fractures/100 person-years. The unadjusted hazard of nonhip skeletal fracture was 2.52 (95% confidence interval 2.05 to 3.12) for subjects with hip fracture compared with subjects without; when adjusted for other known fracture risk factors the hazard ratio was 1.62 (1.30 to 2.02). Men and women had a similar relative risk increase. The increased risk of secondary fracture after hip fracture persisted over time. CONCLUSIONS: A hip fracture is associated with a 2.5-fold increased risk of subsequent fracture, which is not entirely explained by prefracture risk factors. Careful attention to secondary prevention is warranted in these patients. PMID- 14530911 TI - Vitamin D receptor initiation codon polymorphism, bone density and inflammatory activity of patients with ankylosing spondylitis. AB - OBJECTIVES: Osteoporosis is a common finding in ankylosing spondylitis (AS) and may contribute to spinal deformity and bone pain. Bone metabolism as well as inflammatory processes are influenced by the vitamin D receptor gene (VDR). We investigated initiation codon ( FokI) and 3'UTR ( BsmI) polymorphisms of the VDR for whether there could be an association with bone mineral density (BMD) in relation to bone metabolism or inflammatory activity in patients with AS. METHODS: In this study, 104 patients with AS (m/w 71/33, mean age 41+/-12 years) were investigated for their lumbar and femoral BMD by DEXA and in part by QCT measurements and compared to 54 healthy controls. Disease activity indices, serum markers of bone metabolism and inflammation were recorded. FokI and BsmI polymorphisms of the VDR were genotyped using genomic DNA from peripheral leukocytes with present or absent restriction sites defined as alleles " f" and " b" or " F" and " B," respectively. RESULTS: In male AS patients, FokI genotypes were significantly associated with spinal but not with femoral BMD values ( P=0.01) as independent predictors of low BMD, which was also influenced by BMI, and inflammatory and pain indices. CRP and ESR values were also significantly associated with FokI genotypes. BMD in female patients showed no significant association with either FokI or BsmI genotypes of the VDR. CONCLUSION: This is the first evidence that the VDR gene may be involved in BMD differences, bone metabolism and inflammatory processes in ankylosing spondylitis. A possible interaction of the vitamin D system, cytokines and bone could define new diagnostic and therapeutic implications in ankylosing spondylitis. PMID- 14530913 TI - Development of a submerged-liquid sporulation medium for the johnsongrass bioherbicide Gloeocercospora sorghi. AB - Submerged culture experiments were conducted in three phases to determine the optimal medium for rapidly producing conidia of the fungal bioherbicide Gloeocercospora sorghi. In phase I, 18 crude carbon sources were evaluated to determine which would support sporulation. Under the conditions tested, butter bean and lima bean brines (1.5-4.6 mS/cm) provided best conidiation. In phase II, a fractional-factorial design was utilized to screen 76 different medium adjuncts in combination with butter bean brine for improved sporulation. d-Mannitol and carboxymethylcellulose (CMC) were the only acceptable factors that resulted in a significant improvement. In phase III, a central composite design with response surface methodology was used to optimize concentrations of these critical factors. The model predicted optimal sporulation in a medium composed of 2.69 mS/cm butter bean brine +0.043 M d-mannitol +0.37% w/v CMC with an expected titer of 1.51x10(7) conidia/ml. Actual mean titer attained with the model-derived medium was 1.91x10(7) conidia/ml. Optimal sporulation occurred at 25.5 degrees C in this medium and conidia remained viable up to 2.71 days when stored at 12 degrees C. No significant difference was observed in virulence of conidia produced on agar vs washed conidia produced in the model-derived (liquid) medium. PMID- 14530912 TI - Bone mineral density in premenopausal women treated for node-positive early breast cancer with 2 years of goserelin or 6 months of cyclophosphamide, methotrexate and 5-fluorouracil (CMF). AB - The purpose of this study was to compare changes in bone mineral density (BMD) in premenopausal patients with node-positive early breast cancer treated with goserelin (Zoladex) or cyclophosphamide, methotrexate and 5-fluorouracil (CMF). Patients ( n=1640) were randomized to goserelin (3.6 mg every 28 days for 2 years) or CMF (sixx28-day cycles) treatment. In a protocoled sub-study involving 96 patients from eight centers (goserelin: n=53; CMF: n=43), lumbar spine (L2-L4) and femoral neck BMD were assessed by dual X-ray absorptiometry at baseline and then annually for 3 years. At the end of the 2-year goserelin-treatment period, mean BMD losses for goserelin-treated and CMF-treated patients were -10.5% and 6.5% ( P=0.0005) for lumbar spine and -6.4% and -4.5% ( P=0.04) for femoral neck, respectively. At 3 years, partial recovery of BMD was observed in goserelin recipients. In contrast, mean BMD losses for the CMF group indicated persistent BMD loss. No significant differences in BMD were observed between groups at the 3 year assessment of the spine or femoral neck. In the CMF group, based on amenorrhea status at 48 weeks, BMD losses at the lumbar spine were greater for amenorrheic than non-amenorrheic patients. Ovarian suppression resulting in amenorrhea was closely related to BMD loss in both treatment groups. Overall, patients who received CMF did not show recovery of BMD throughout follow-up, whereas partial recovery was observed 1 year after cessation of goserelin therapy, associated with the return of ovarian function in the majority of patients. PMID- 14530915 TI - Cell and tissue behavior on micro-grooved surfaces. AB - In this review, we discuss substrates and implant surfaces provided with micrometer-sized groove and ridge patterns. Such "microgrooves" influence cell behavior: the cells align themselves, and migrate guided by the surface grooves. This phenomenon is known as "contact guidance". First, cell structure and cell attachment behavior are described. Then techniques for the production of microgrooves are addressed, and a summary is given of a number of previous in vitro and in-vivo experiments on this subject. Based on the knowledge of cell movement, we suggest a theory involving the dynamics of fibrous cellular components in the filopodium. Finally, future directions for this type of research, and implications for medical and dental implantology, are addressed. PMID- 14530916 TI - Carcinogenicity of bisphenol-A in Fischer rats and B6C3F1 mice. AB - Bisphenol-A (BP-A; 4,4'-isopropylidenediphenol) is a monomer of plastics commonly used in various consumer products, and is used as an intermediate in the manufacture of epoxy, polycarbonate, and polyester-styrene resins. A National Toxicology Program carcinogenesis bioassay of BP-A (>98% pure) was conducted by feeding diets containing 0, 1000, or 2000 ppm BP-A to groups of 50 male and 50 female Fischer (F)344 rats; 0, 1000, or 5000 ppm to groups of 50 male B6C3F1 mice; and 0, 5000, or 10,000 ppm to groups of 50 female B6C3F1 mice for 103 weeks. The mean body weights of the low- and high-dose rats and of female mice and high-dose male mice were lower than those of the controls throughout much of the study. Lower body weight gains in rats were likely caused by reduced food consumption. Survivals were comparable among groups. Regarding neoplasia, leukemias occurred at increased incidences in BP-A-dosed rats of both sexes: male, 13/50 controls vs 12/50 low-dose and 23/50 high-dose (P < 0.03); in females, the respective findings were 7/50, 13/50, and 12/50. Interstitial-cell tumors of the testes were increased in BP-A-dosed male rats: 35/49 controls vs 48/50 (P < 0.01) and 46/49 (P < 0.01); and an increasing trend was observed for mammary gland fibroadenomas in male rats (P < 0.05, 0/50 controls vs 0/50 and 4/50). In male mice, lymphomas/leukemias were increased: 2/49 controls vs 9/50 (P < 0.05) and 5/50. Multinucleated giant hepatocytes were observed in male mice (1/49 controls vs 41/49 and 41/50), whereas there was no increase of liver tumors. In their BP-A bioassay report, the National Toxicology Program concluded that there was no convincing evidence that BP-A was carcinogenic for rats or mice. However, the marginal increases in leukemias in male and female rats, along with increases in the combined incidence of lymphomas and leukemias in male mice, suggest that BP-A may be associated with increased cancers of the hematopoietic system. Increases in interstitial-cell tumors of the testes in rats were also evidence of carcinogenesis, as was the unusual occurrence of mammary gland fibroadenomas in male rats. PMID- 14530917 TI - Effect of vagal afferent conditioning on trigeminal motor neuron activity associated with tooth-pulp-evoked jaw-opening reflex. AB - To clarify the vagal afferent modifying effect on the neurons constituting the nociceptive jaw-opening reflex (JOR), we conducted extracellular recording of trigeminal motor nucleus (TMN) neuron activity in pentobarbital-anesthetized rats. Of 12 TMN neuron responses evoked by tooth pulp (TP) stimulation, 10 were suppressed by vagal afferent conditioning (83%). The mean time of the suppressive effect on the TMN neurons paralleled that found with the digastric electromyogram (dEMG), and maximal inhibition of both TMN neuronal spikes and the dEMG amplitude were observed at a 50-ms conditioning-test interval. The ratio of inhibition was approximately 38%. Seven of 12 units were activated antidromically by digastric muscle stimulation (1-3 mA, 0.1 ms, 2 Hz) and fulfilled the criteria for antidromic activation. Vagal afferent conditioning stimulation had no effect on the antidromic TMN neuronal responses to digastric muscle stimulation. These results suggest that suppression of the TP-evoked JOR in response to vagal afferent stimulation in rats is the result of an inhibitory effect on the sensory neurons rather than on the motor neurons. PMID- 14530918 TI - Sexual dimorphism of porcine amelogenins: male-specific amelogenins have strong adsorption properties onto apatite crystals. AB - The present studies were undertaken to investigate the sexual dimorphism of porcine amelogenins and to gain information as to whether excesses of male amelogenins, if any, possess functional significance in protein-crystal interactions. Enamel proteins, including the intact full-length amelogenins and their degraded polypeptides, were isolated from the secretory enamel of male and female pigs. To identify the amelogenins among the separated pools of male- and female-matrix proteins, rabbit anti-C13 and C25 peptide sera were used, which reacted specifically with the conserved C-terminal domain. Immunoblotting showed that a few extra members of the amelogenins, sharing common epitopes at the C terminus, were recognized in male products. The apparent yield of the male amelogenins was only marginal, on the basis of their stained intensities on the gel, but the secreted male amelogenins demonstrated selective (probably the strongest among the amelogenins) adsorption properties onto apatite crystals. Reflecting the general symmetric electrophoretic profiles of the male- and female enamel proteins in toto, there were no sex-linked differences in the protein crystal interaction and the resulting regulatory function of crystal precipitation. PMID- 14530919 TI - Ultrastructural study of hormonally responsive striated duct cells in the mouse sublingual gland. AB - In semithin sections stained with Heidenhain's iron hematoxylin, a few scattered granular cells were observed in the striated ducts (SDs) of sublingual glands (SLGs) of the mouse; they were seen normally only in the glands of adult males. However, it was shown by electron microscopy that many SD cells, other than these granular cells, had apical secretory granules, thus forming a granular striated tubule (named the GST in this study) in a portion of SD segments in both sexes. Sublingual GST cells had very small dense secretory granules near the apical surface, with the nucleus in the apical one-third to one-half of the cell; small Golgi apparatus; sparse rough endoplasmic reticulum (RER); and well-developed basal infoldings. However, some granular cells in male GSTs had abundant large dense secretory granules in the apical two-thirds of the cell, a basal nucleus, and modest basal infoldings. Such granular SD cells disappeared after castration in males. Granular SD cells could be induced in the GSTs of females by the injection of 5 alpha-dihydrotestosterone (DHT), triiodothyronine (T(3)), and/or dexamethasone (Dex); given simultaneously, these hormones acted synergistically in this induction. These results indicate a close similarity between the duct systems of the SLG and those of the submandibulan gland (SMG) of the mouse: granular SD cells of the GST in the SLG resemble GCT cells in the SMG in expressing some of the same biologically active polypeptides, in being sexually dimorphic, and in being under the same multihormonal regulation. PMID- 14530920 TI - Effect of filler size on wear resistance of resin cement. AB - The purpose of this study was to evaluate the effect of filler size on the wear of resin cements. Materials tested included four experimental dual-cure resin cements (Kuraray) consisting of different-sized filler particles. A rectangular box cavity was prepared on the flattened occlusal surface of extracted human molars. Ceramic inlays for the cavities were fabricated using the Cerec 2 system. The Cerec inlays were cemented with the respective cements and adhesive systems according to the manufacturer's directions. The restored surface was finished by wet-grinding with an 800-grit silicon carbide paper. Six specimens were prepared for each resin cement. Half of the specimens were subjected to a three-body wear test for 200,000 cycles, and the others were subjected to a toothbrush abrasion test for 30,000 cycles. The worn surface of each restoration was scanned by a profilometer (Surfcom 475 A) at eight different points for each restoration. The wear value was determined by measuring the vertical gap depth on the profilometric tracings. The data were statistically analyzed by one-way analysis of variance (ANOVA) and Scheffe's test. The results showed that, with increase of filler size, the wear value decreased in the toothbrush test and increased in the three-body wear test. The cement with 0.04-microm filler exhibited the lowest wear value among the materials in the three-body wear test, and the same wear value as the cement with 0.97-microm filler in the toothbrush test. Based upon the results of this study, it is concluded that the wear of resin cements was affected by the filler size as well as the mode of wear test. PMID- 14530921 TI - Tensile bond strength of light-cured resin-reinforced glass ionomer cement with delayed light exposure. AB - In the clinical situations, the time intervals between material mixing and light exposure during bracket bonding, using light-cured resin-reinforced glass ionomer cement (LCGIC), may vary for each individual bracket. This study determined the tensile bond strengths of LCGIC subjected to various time intervals, and evaluated the durability with thermocycling. Comparisons were made between LCGIC and light-cured composite resin (LCR). Two hundred and forty bovine teeth were chosen as specimens. Light exposure was performed 5, 10, 20, and 40 min after the commencement of powder/liquid mixing. The durability was evaluated by thermocycling for 2000 times at temperatures between 5 degrees C and 55 degrees C, with a 30-s dwell time. Tensile bond strengths of LCGIC and LCR after 5 min, representing the general condition in clinical use, equaled 5.7 +/- 1.5 MPa and 5.1 +/- 2.6 MPa, respectively. For the LCGIC groups, no significant differences were seen between bond strengths with and without thermocycling. Also, no significant differences were noted among any time intervals. For the LCR groups, there were also no significant differences with and without thermocycling. The tensile bond strength of LCR showed highly significant differences within groups across time. Compared with LCR, the failure sites for brackets bonded with LCGIC appeared to be predominantly at the bracket/adhesive interface. The standard deviations of LCR were high when compared with those of LCGIC. The bond strength of LCGIC with or without thermocycling surpassed the clinically required minimum. LCGIC may be an advantageous alternative to LCR for orthodontic bracket bonding. PMID- 14530922 TI - The effect of bolus size on the chewing cycle in humans. AB - No general agreement exists regarding the effect that bolus size has on masticatory movement, probably because both the size and texture of food change during mastication. In this experiment, in order to clarify the effect of bolus size on masticatory movement, a food that does not change in size and texture- chewing gum--was chosen, and the relationship between bolus size and the chewing cycle was analyzed. Ten healthy subjects in their twenties were asked to chew pieces of softened chewing gum of four different sizes. For ten cycles, beginning with the fifth cycle of mastication, gape and masticatory width were calculated for the spatial parameter of the chewing cycle, and cycle time was calculated as the temporal parameter. The relationship between these parameters and the bolus size was investigated. As the bolus size increased, the spatial and temporal parameters increased. In addition, there was a positive correlation between the bolus size and each parameter. The influence of the bolus size was as follows: gape, r = 0.91; masticatory width, r = 0.79; and cycle time, r = 0.74 (all, P < 0.001). From these results it was concluded that the shape of the chewing cycle was altered by the size of the food bolus, and that the changes in sensory input from the peripheries greatly affected the masticatory movement. PMID- 14530923 TI - Analyzing the eye movement of dentists during their reading of CT images. AB - In order to evaluate the eye movements of dentists when they were interpreting radiographs, ten normal computed tomography (CT) images and ten images with pathologic lesions were shown to eight dentists, and the pattern of their eye movement was qualitatively analyzed. Six fixation point parameters were calculated, including the time required to discriminate between normal and pathologic images (X(1)), the total fixation point count (X(2)), the total travel distance between fixation points (X(3)), the average time spent on each fixation point (X(4)), the total gaze fixation time (X(5)), and the maximum gaze fixation time spent on each image (X(6)). When the subjects were interpreting pathologic images, X(2) and X(4) were shorter; however, when they were viewing normal images, time was spent on observing multiple fixation points before the completion of the interpretive process. While pathologic images were recognized through top-down processing, there was a tendency for normal images to be recognized through bottom-up processing. The results of discriminant analysis, using a linear discriminant function, indicated that the independent variables X(2) and X(4) and the dependent variable X(5) were the only variables that contributed significantly to differentiating between normal and pathologic images. The linear discriminant function was Z = 9.0 x 10(-2) x X(2) + 3.0 x X(4) - 2.1 (discriminant score: Z < 0, pathologic image; Z > or = 0, normal image). When the mean value of each individual's gaze fixation data was substituted into the discriminant formula, the hit rate for normal and pathologic images was discriminated at 94% (15 of 16). PMID- 14530924 TI - Transcortical-transventricular approach in colloid cysts of the third ventricle: surgical experience with 26 cases. AB - Colloid cysts of the third ventricle account for 0.5-2% of all intracranial tumors. The treatment of these benign tumors remains controversial, and the best surgical option has not yet been determined. Between 1995 and 2002, 27 patients with colloid cysts of the third ventricle presented at our clinic. Twenty-six underwent transcortical-transventricular approaches. One refused surgical treatment. There was no surgical mortality. The main morbidity was epileptic seizures in two patients. Overall outcome was good in all patients. The mean follow-up period was 3.4 years. There were no tumor recurrences. The transcortical-transventricular approach can be used safely to excise third ventricle colloid cysts with low risk of mortality and morbidity. PMID- 14530926 TI - Novel splice site CACNA1A mutation causing episodic ataxia type 2. AB - Episodic ataxia type 2 (EA-2) is an autosomal dominant neurological disorder, characterized by episodes of ataxia, vertigo, nausea, nystagmus, and fatigue, associated with acetazolamide responsiveness. The disease is caused by mutations in the P/Q-type calcium channel Ca(v)2.1 subunit gene, CACNA1A, located on chromosome 19p13.2. We analyzed a family with 13 affected individuals for linkage to this locus and reached a two-point maximum LOD score of 4.48. A novel CACNA1A mutation, IVS36-2A>G, at the 3' acceptor splice site of intron 36 was identified by sequencing. It is the first described CACNA1A acceptor splice site mutation and the most C-terminal EA-2-causing mutation reported to date. PMID- 14530927 TI - Molecular dynamics study of 4-OH-phenylacetyl- D-Y(Me)FQNRPR-NH2 selectivity to V1a receptor. AB - G protein-coupled receptors relay diverse extracellular signals into cells via a common mechanism, involving activation of cytosol G proteins. The mechanism underlies the actions of approximately 50% of all drugs. In this work, we focus on simulating three protein-ligand complexes of the neurohypophyseal hormone analog 4-OH-phenylacetyl- D-Y(Me)FQNRPR-NH2 (I) with the human V1a, V2 and oxytocin receptors. The peptideI is a potent selective V1a receptor antagonist. To obtain relaxed models of the complexes, the following techniques were used: docking ofI into the vasopressin V1a, V2 and oxytocin receptor models, optimization of the geometry of the resulting complexes and molecular dynamics in a fully hydrated 1-palmitoyl-2-oleoyl- sn-glycero-3-phosphatidylcholine lipid bilayer. The results of the simulations allow us to draw some conclusions about the ligand selectivity to V1aR. PMID- 14530928 TI - Molecular recognition between 4a S/R-galanthamine diastereoisomers and alpha cyclodextrin. AB - Molecular recognition between 4a S/R-galanthamine diastereoisomers (1: 4a S galanthamine;2: 4a R-galanthamine) and alpha-cyclodextrin (alpha-CD) were studied by use of docking and molecular dynamics (MD) simulation approaches. The binding energy of constructed 2...alpha-CD complexes is approximately 17 kcal mol(-1) lower than that of 1...alpha-CD, implying a stronger binding ability of 2 with alpha-CD than that of1. The theoretical modeling result is consistent with our previous CZE result, which demonstrated that alpha-CD is an efficient chiral additive for separating 1 and 2. The modeling result also indicates that both hydrophobic interaction and H-bond force may work as major factors for molecular recognition between the galanthamine diastereoisomers and alpha-CD. PMID- 14530929 TI - Fungal colonization of shrub willow roots at the forefront of a receding glacier. AB - Shrub willows ( Salix spp.) form associations with arbuscular mycorrhizal (AM), ectomycorrhizal (EM) and dark septate endophytic (DSE) fungi. Willow root colonization by these three types of fungi was studied on a deglaciated forefront of Lyman Glacier, Washington, USA. Root colonization was low; less than 1% of the root length was colonized by AM and 25.6% by DSE. EM colonized 25% of the root tips and 19.4% of the root length. AM and DSE colonization were not related to distance from the present glacier terminus or to canopy cover. EM colonization increased with distance from the glacier terminus based on gridline intercept data but not on root tip frequency data. Availability of propagules in the substrate was low, but numbers of propagules increased with distance from the glacier terminus. The EM communities were dominated by three ascomycetes showing affinity to Sordariaceae in BLAST analyses. Other frequent taxa on the glacier forefront included species of Cortinariaceae, Pezizaceae, Russulaceae, Thelephoraceae and Tricholomataceae. When occurrence of individual taxa was used as a response variable to canopy cover, distance from the glacier terminus, and their interaction, four different fungal guilds were identified: 1) fungi that did not respond to these environmental variables; 2) fungi that occurred mainly in intercanopy areas and decreased with distance from the glacier terminus; 3) fungi that were insensitive to canopy cover but increased with distance from the glacier terminus; 4) fungi that occurred mainly under willow canopies and increased with distance from the glacier terminus. We suggest that fungal colonization is mainly limited by fungal propagule availability. Environmental conditions may also limit successful establishment of plant-fungus associations. We propose that the four EM guilds partly explain successional dynamics. The initial EM community comprises fungi that tolerate low organic matter and nitrogen environment (first and second guilds above). During later community development, these fungi are replaced by those that benefit from an increased organic matter and nitrogen environment (third and fourth guilds above). PMID- 14530930 TI - Local cortical cerebral blood flow and response to carbon dioxide during anesthesia in patients with moyamoya disease. AB - PURPOSE: The CO(2) reactivity of cortical cerebral vessels and local cortical blood flow (l-CoBF) were evaluated during anesthesia in patients with moyamoya disease who were undergoing revascularization surgery. METHODS: Using laser Doppler flowmetry, the CO(2) reactivity of cortical cerebral vessels and l-CoBF were measured continuously in five patients at the local surgical field of the middle cerebral artery (MCA) territory. RESULTS: Local-CoBF values obtained during the normocapnic condition varied from site to site of gyrus in the MCA region (0-73 ml. 100 g(-1).min(-1)). Local-CoBF was maximal at 39-43 mmHg of the PaCO(2) range, and decreased above and below this range. The response of l-CoBF to CO(2) was larger at the sites where the maximal level was obtained during normocapnia. In two patients, l-CoBF decreased by about 50%, and remained law even 40 min after administration of acetazolamide. CONCLUSION: In patients with moyamoya disease, l-CoBF values obtained during the normocapnic condition varied from site to site of gyrus, and not only hypocapnia but also hypercapnia decreased l-CoBF within the MCA region. PMID- 14530931 TI - Cuffed oropharyngeal airway and capnometry: comparison of end-tidal and arterial carbon dioxide pressures. AB - PURPOSE: The aim of this study was to investigate the reliability of end-tidal CO(2) tension (PetCO(2)) as a predictor of PaCO(2) during anesthesia in patients breathing spontaneously via a cuffed oropharyngeal airway (COPA). METHODS: Twenty adult patients scheduled for minor surgery were included in this study. After propofol injection, an appropriate size of COPA was inserted. Anesthesia was maintained with 60% nitrous oxide in oxygen (total flow rate of 5 l.min(-1)) supplemented with propofol infusion. The patients were allowed to breathe spontaneously throughout the procedure. PaCO(2) and PetCO(2) were simultaneously measured when a steady state of anesthesia was reached. RESULTS: PaCO(2) (48.8 +/ 5.4 mmHg, range 36.2-58.0 mmHg) was higher than PetCO(2) (43.1 +/- 4.2 mmHg, range 32-51 mmHg) in all patients. The difference between end-tidal and arterial CO(2) tension was 5.7 +/- 3.2 mmHg (range 0.5-13.0 mmHg), and was significantly correlated with PaCO(2) ( P < 0.01). CONCLUSION: The results of this study suggest that PetCO(2) in anesthetized patients breathing spontaneously through a COPA is sometimes unreliable as an indicator of PaCO(2) level, and there is some possibility of unexpected hypercapnia. PMID- 14530932 TI - Prophylactic antiemetic therapy with droperidol in patients undergoing laparoscopic cholecystectomy. AB - PURPOSE: The incidence of postoperative nausea and vomiting (PONV) following laparoscopic cholecystectomy (LC) is relatively high when no prophylactic antiemetic is given. We have studied the efficacy of a commonly used and well established antiemetic, droperidol, for the prevention of PONV in patients undergoing LC. METHODS: In a randomized, double-blind, placebo-controlled study, 60 patients received placebo (saline) or droperidol 50 microg.kg(-1) (maximum dose, 2.5 mg) intravenously immediately before the induction of anesthesia (n = 30 of each). A standard general anesthetic technique was employed throughout. RESULTS: A complete response, defined as no PONV and no need for another rescue antiemetic medication during the first 24 h after anesthesia, was 57% and 83% in patients who had received placebo and droperidol 50 microg.kg(-1), respectively ( P < 0.05). No clinically serious adverse events were observed in any of the groups. CONCLUSION: Prophylactic antiemetic therapy with droperidol 50 microg.kg( 1) (maximum dose, 2.5 mg) is highly effective for preventing PONV after LC. PMID- 14530933 TI - Expression of heat shock protein 70 mRNA in polymorphonuclear cells responding to surgical stress. AB - PURPOSE: This study was performed to investigate the expression of heat shock protein (HSP) 70 mRNA in polymorphonuclear neutrophils (PMN) as a possible new biomarker for surgical stress. METHODS: The HSP70 mRNA in PMN of 10 patients who underwent lobectomy was evaluated by Northern blot analysis. Their leukocyte counts, including white blood cells (WBC) and PMN, plasma cortisol levels, and plasma interleukin-6 (IL-6) levels, were obtained by cell counting, radioimmunoassay, and enzyme-linked immunosorbent assay, respectively. RESULTS: The level of HSP70 mRNA in PMN slightly increased at the end of surgery and showed a significant increase 6 h after surgery. It promptly decreased at 24 h postoperatively and returned to the basal preanesthetic level 48 h after surgery. On the other hand, WBC/PMN counts, plasma cortisol, and IL-6 significantly increased at the end of surgery. WBC/PMN counts remained at increased levels until 48 h postoperatively. Cortisol peaked at 6 h postoperatively and gradually decreased. IL-6 reached a maximum at 1 h postoperatively, then tapered down to its basal level at 48 h postoperatively. CONCLUSION: Expression of HSP70 mRNA in PMN that is induced after thoracic surgery appears to be a promising candidate as a marker for evaluating surgical stress. PMID- 14530934 TI - Epidural block during general anesthesia attenuates urinary trypsin inhibitor excretion in lower abdominal surgery. AB - PURPOSE: The aim of this study was to elucidate whether urinary trypsin inhibitor excretion differs between general anesthesia (GA) and epidural block during general anesthesia (EPI) in lower abdominal surgery. METHODS: Sixteen women undergoing abdominal total hysterectomy were assigned to the GA and EPI groups. The GA group received propofol induction and maintenance with isoflurane, nitrous oxide, and vecuronium. The EPI group received epidural block, followed by propofol induction and maintenance with isoflurane and nitrous oxide. The levels of adrenocorticotropic hormone and cortisol during anesthesia and on postoperative days 1, 2, and 3, and the levels of urinary trypsin inhibitor in 12 h urine from the day of surgery to postoperative day 3, were measured. RESULTS: As compared with the EPI group, the GA group had a higher level of adrenocorticotropic hormone at the completion of anesthesia, higher levels of cortisol at the completion of anesthesia and postoperative day 2, and higher excretion of urinary trypsin inhibitor on the day of surgery and postoperative days 1 and 2. CONCLUSION: The present results suggest that excretion of urinary trypsin inhibitor into the urine under epidural block during general anesthesia is lower than that under general anesthesia alone in lower abdominal surgery. This is probably due to the difference in endocrine response to surgery between the two types of anesthesia. PMID- 14530935 TI - Effects of sevoflurane anesthesia on plasma inorganic fluoride concentrations during and after cardiac surgery. AB - PURPOSE: Sevoflurane metabolism results in the production of inorganic fluoride, which is known to be nephrotoxic. Since marked changes in body temperature and hemodynamics in cardiac surgery affect sevoflurane metabolism, plasma inorganic fluoride concentrations may differ in this situation compared with other types of surgery. We therefore measured plasma inorganic fluoride concentrations during and after sevoflurane anesthesia in patients undergoing cardiac surgery. METHODS: Sixteen patients undergoing coronary artery bypass grafting or valve replacement were premedicated with 5-10 mg midazolam and 0.5 mg scopolamine injected intramuscularly. Anesthesia was induced with 5-10 mg midazolam, 0.5-1 mg fentanyl, and 0.12-0.15 mg.kg(-1) vecuronium. Following tracheal intubation, anesthesia was maintained with oxygen, sevoflurane, and fentanyl. At the onset of cardiopulmonary bypass (CPB), sevoflurane was discontinued, and additional fentanyl, midazolam, and pancuronium were administered. Plasma inorganic fluoride concentrations were measured before anesthesia, immediately before and after CPB, and at 0, 2, 6, 12, 24, and 48 h after anesthesia. RESULTS: The individual maximum plasma inorganic fluoride concentration was 19.2 +/- 7.2 micromol.l(-1) (mean +/- SD; range, 9.2-36.7). The mean plasma inorganic fluoride concentrations increased during anesthesia, but the rate of increase decreased after the initiation of CPB. Concentrations peaked at 2 h after anesthesia and decreased thereafter. The concentrations in three cases continued to increase 2 h after anesthesia. CONCLUSION: The plasma inorganic fluoride concentrations observed in patients undergoing cardiac surgery were below nephrotoxic levels. However, the decrease in mean fluoride concentration after anesthesia was slower than that in the previous study in general surgical patients. PMID- 14530936 TI - The role of skeletal muscle and liver on lactate metabolism during hypoxia in rats. AB - PURPOSE: This study was planned to investigate whether the skeletal muscle and liver produce or consume lactate under hypoxic conditions. METHODS: Wister rats were anesthetized and mechanically ventilated. Microdialysis probes were inserted into the rat skeletal muscle and liver, and arterial cannulation was performed. Hypoxia was induced for 30 min by the inhalation of 10% oxygen in nitrogen. Interstitial lactate concentrations in the skeletal muscle and liver were measured using an in vivo microdialysis method before, during, and after hypoxic hypoxia. The blood lactate concentration, mean arterial blood pressure, and blood gas were also measured. RESULTS: Before hypoxia, there was no significant difference among the blood lactate concentration and interstitial lactate concentrations of the skeletal muscle and liver. During hypoxia, arterial oxygen tension decreased to 34.2 +/- 1.3 mmHg, and the lactate concentrations in these tissues increased significantly in comparison to the control values. However, the lactate concentrations in the skeletal muscle and liver interstitium were significantly lower than that in the blood, with the peak lactate concentration in the skeletal muscle interstitium being only one-third of that in the blood. After correction of hypoxia, the blood lactate concentration decreased to levels comparable to the skeletal muscle and liver interstitial lactate concentrations. CONCLUSION: It is suggested that the skeletal muscle as well as the liver may consume lactate under hypoxic hypoxia. PMID- 14530937 TI - Low-flow anesthesia: theory, practice, technical preconditions, advantages, and foreign gas accumulation. PMID- 14530938 TI - A reliable method for preventing pain on injection of propofol. PMID- 14530939 TI - Anesthetic management for cesarean section in a mother after the Fontan procedure. PMID- 14530940 TI - Malignant hyperthermia triggered by isoflurane and suxamethonium in a patient who underwent apparently uneventful halothane anesthesia previously: a case report. PMID- 14530941 TI - A case of aortic dissection without severe chest pain during an attempt at epidural anesthesia. PMID- 14530942 TI - Improved oxygen delivery to the fetus during cesarean section under sevoflurane anesthesia with 100% oxygen. AB - PURPOSE: To assess the potential benefits of sevoflurane with 100% oxygen in cesarean section in terms of oxygen delivery to the fetus, neonatal depression, and uterine contractility. METHODS: Thirty-six patients undergoing elective cesarean section were enrolled. After thiamylal induction, 0.7% sevoflurane-60% nitrous oxide-40% oxygen anesthesia was administered in group G1 (n = 9), and 1.7% sevoflurane-100% oxygen anesthesia was administered in group G2 (n = 9). Spinal anesthesia under oxygen nasal prong was used in group SP (n = 18). RESULTS: At delivery, the PO(2) values in the maternal artery and the umbilical vein and artery (MA, UV, UA) of group G2 (474 +/- 50, 43 +/- 9, 32 +/- 9 mmHg, respectively) were significantly greater than those in groups G1 (228 +/- 46, 31 +/- 4, 23 +/- 5 mmHg, respectively) and SP (147 +/- 21, 30 +/- 7, 18 +/- 7 mmHg, respectively). The SO(2) in the UA of group G2 (56 +/- 17 %) was also greater than that in groups G1 (34 +/- 10 %) and SP (32 +/- 10 %). The sevoflurane concentrations at delivery in the MA, UV, and UA in group G2 were almost threefold higher than those in group G1, whereas all the newborns in the three groups had Apgar scores of 8 or more at 5 min, and the intraoperative blood loss did not differ among the groups. CONCLUSION: Sevoflurane anesthesia with 100% oxygen in elective cesarean delivery improves oxygen delivery to the fetus without severe neonatal depression, prolonged uterine relaxation, or increased blood loss. PMID- 14530943 TI - Effects of nitroglycerin on fractal features of short-term heart rate and blood pressure variability. AB - PURPOSE: We investigated the effect of nitroglycerin(NTG) on fractal features of short-term heart rate variability (HRV) and blood pressure variability (BPV) using coarse-graining spectral analysis (CGSA). METHOD: Nine healthy young volunteers participated in this study. Five-minute recordings of electrocardiogram and blood pressure estimated by photoplethysmograph were made during stepwise NTG infusions of 0.2, 0.4, 0.8, and 1.6 microg.kg(-1).min(-1) under rate-controlled breathing at 0.25 Hz. CGSA broke down the total power of the time series into harmonic (low- and high-frequency) and nonharmonic (beta of 1/f(beta) and %fractal) components. RESULTS: A statistically significant difference from the control period was observed during the maximum dose of NTG infusion, with decrease in mean blood pressure, shortening of mean R-R interval, and increase in plasma norepinephrine and epinephrine. The beta in HRV increased significantly (0.89 +/- 0.06 vs. 1.27 +/- 0.13, P < 0.05). However, %fractal was not affected (47.9 +/- 6.7 vs. 50.1 +/- 4.0). Indicators of parasympathetic and sympathetic nervous system activity showed reduced and increased values, respectively. No change in BPV was observed for any measurement. CONCLUSION: The data suggest that NTG significantly affected fractal features, as well as harmonic components, of short-term HRV. NTG had no effect on BPV, suggesting a different mechanism for genesis of 1/f(beta) fluctuation in BPV and HRV. PMID- 14530944 TI - Comparative effects of etomidate, ketamine, propofol, and fentanyl on myocardial contractility in dogs. AB - PURPOSE: The present study was carried out to determine the direct effects of etomidate, ketamine, propofol, and fentanyl on myocardial contractility, and whether fentanyl would enhance the myocardial depression caused by propofol. METHOD: The anesthetics were injected directly into the circuit that supplied blood to the left circumflex coronary artery (LCX) in anesthetized open-chest dogs. Myocardial contractility was evaluated from measurements of percent segmental shortening (%SS). RESULTS: Etomidate, ketamine, and propofol significantly reduced %SS in a dose-dependent manner. The %SS values with 1.6 and 3.2 mg of etomidate were similar to those with 3.2 and 6.4 mg of ketamine, respectively, and the %SS value with 6.4 mg of propofol was similar to those with 3.2 and 6.4 mg of ketamine. Fentanyl alone had no effects on myocardial performance and did not influence the effect of propofol on %SS. CONCLUSION: On the basis of clinical doses, the direct myocardial depressant effect of ketamine is more than twice as potent as that of etomidate and slightly more than that of propofol. Fentanyl has no inotropic effect and does not enhance the direct myocardial depressant effect of propofol. PMID- 14530945 TI - Inhibitory effect of nitric oxide on the metabolism of halogenated volatile anesthetics by cytochrome P-450. AB - PURPOSE: This study was undertaken to examine the effect of nitric oxide (NO) on the metabolism of halogenated volatile anesthetics (HVA) by cytochrome P-450 (CYP) under both aerobic and anaerobic conditions using rat hepatic microsomes. METHODS: A microsomal fraction was prepared by centrifugation from normal and phenobarbital-treated male SD rats. The anaerobic metabolism of HVA by CYP was followed by measuring the formation of a halothane CYP complex spectrophotometrically. Aerobic CYP activity was determined using either the defluorination of sevoflurane or the demethylation of aminopyrine. RESULTS: The formation of the halothane-CYP complex was dose-dependently inhibited by NO. NO also decreased CYP defluorination of halothane in a dose-dependent manner. In phenobarbital-induced microsomes, the inhibition rates of both complex formation and the defluorination of halothane were the same as those seen in normal microsomes. Although the defluorination of sevoflurane and the demethylation of aminopyrine were inhibited by NO aerobically, the inhibition was much less than that of the metabolism of halothane under anaerobic conditions. CONCLUSION: These results suggest that NO binds the heme of CYP and inhibits the metabolism of HVA, with the effect lasting for a prolonged period of time. Furthermore, the balance between NO and O(2) is important for NO to inhibit CYP. PMID- 14530946 TI - Role of reactive oxygen in phospholipase A2 activation by ischemia/reperfusion of the rat kidney. AB - PURPOSE: To investigate the role of phospholipase A(2) (PLA(2)) in reperfusion injury of the kidney in an in vivo animal model, renal mitochondrial PLA(2) activity was measured under three different conditions. METHODS: Male Wistar rats (n = 72) anesthetized with pentobarbital underwent renal ischemia surgically for 45 min and were reperfused for the indicated time (renal ischemia/reperfusion). Treatments included reperfusion for various predetermined periods (phase 1), exposure to hyperbaric oxygen (phase 2), and administration of reactive oxygen species (ROS) scavenger (phase 3). Thereafter, each kidney was harvested, and mitochondrial PLA(2) activity was measured by a radioisotope technique. RESULTS: Ischemia/reperfusion resulted in time-related PLA(2) activation in the renal mitochondria up to 48 h of reperfusion after renal ischemia. Renal mitochondrial PLA(2) activity was further augmented by hyperbaric oxygen exposure prior to reperfusion, whereas administration of the ROS scavengers suppressed mitochondrial PLA(2) activity. CONCLUSION: These data suggest that ROS may play an important role in the in vivo activation of PLA(2) associated with renal ischemia/reperfusion. PMID- 14530947 TI - Does adenosine release taurine in the A1-receptor-rich hippocampus? AB - PURPOSE: To determine whether taurine would be protective in the hippocampus (rich in A1 receptors but devoid of the A2 receptor effector adenylate cyclase, which mediates the release of taurine) by determining whether taurine is released by adenosine in the non-ischemic hippocampus. METHODS: Microdialysis of probes, inserted stereotactically into the bilateral hippocampi of five nonischemic brains of anesthetized rabbits and combined with high-performance liquid chromatography, was used to evaluate the effects of adenosine on taurine concentration in the interstitial fluid. RESULTS: Sequential increase of the dialytic artificial cerebrospinal fluid concentration of adenosine (to 1 x 10(-4) M, 5 x 10(-4) M, 1 x 10(-3) M, 5 x 10(-3) M, and 1 x 10(-2) M) linearly and significantly ( P < 0.0001) increased taurine concentration in the interstitial fluid. CONCLUSION: Adenosine delivered by microdialysis into the interstitial fluid of nonischemic rabbit hippocampus induces release of taurine in a dose dependent manner, suggesting involvement of additional mechanisms besides the already known A2-receptor-induced release. Taurine can be anticipated to have protective effects in the hippocampus as well. PMID- 14530948 TI - Effects of hypothermia on c-fos and zif/268 gene expression following rat forebrain ischemia. AB - PURPOSE: To determine the effects of prophylactic mild hypothermia against transient forebrain ischemia, we examined the levels and localization of c-fos and zif/268 gene expression in the rat hippocampal formation at an early period during reperfusion. METHODS: For the histochemical evaluation, Wistar rats were divided into three groups: normothermia (37 degrees C) during ischemia (n = 5), hypothermia (34 degrees C) during ischemia (n = 5), and sham-operated controls (n = 5). The former two groups were subjected to 10-min forebrain ischemia using the bilateral carotid artery occlusion with arterial hypotension (35 mmHg) technique. After 60 min of reperfusion, the brains were removed and the in situ hybridization technique was used to detect c-fos and zif/268 mRNA expression. Additionally, to determine the histopathological changes of neuronal degeneration, animals were treated with hypothermia during ischemia (n = 4) and normothermia during ischemia (n = 4). They were allowed to survive for 7 days, and then the sections of hippocampal formation were examined by light microscopy. RESULTS: Transient ischemia increased c-fos and zif/268 mRNA signal densities from 1.8- to 6.1-fold compared with that in controls in the hippocampus. Mild hypothermia significantly inhibited the induction of c-fos and zif/268 (P < 0.01). In the CA1 subfield, hypothermia protected against delayed neuronal degeneration (P < 0.05), which was observed after normothermic ischemia. CONCLUSION: The induction of transcription factor-related immediate early genes (IEGs) was a sensitive marker of the cellular response to ischemia. Mild hypothermia uniformly inhibited the induction of IEGs in the hippocampus, but the mechanisms underlying the protective effect of hypothermia against neuronal degeneration cannot be explained only by the IEGs. PMID- 14530949 TI - Interaction of ketamine with mu2 opioid receptors in SH-SY5Y human neuroblastoma cells. AB - PURPOSE: Ketamine is known to interact with opioid receptors. However, because this agent does not produce opioid-like respiratory depression, it might not interact with mu(2) opioid receptors. Therefore, we have studied the interaction of ketamine with mu(2) opioid receptors expressed in SH-SY5Y cells. METHODS: SH SY5Y cells (passage 70-80) were used to obtain ketamine dose-response curves for inhibition of 0.4 nM [(3)H][D-Ala(2),MePhe(4),Gly(ol)(5)] enkephalin (DAMGO) binding to mu(2) opioid receptors and of forskolin (1 microM)-stimulated cyclic AMP (cAMP) formation. RESULTS: Ketamine displaced [(3)H]DAMGO binding in SH-SY5Y cells with a K(i) of 12.1 microM. However, this concentrations did not inhibit forskolin-stimulated cAMP formation, although at supraclinical concentrations, significant inhibition was observed with an estimated IC(50) of 700 microM. CONCLUSION: The present study indicates that a clinically relevant concentration of ketamine interacts with mu(2) opioid receptors. However, no agonist activity was observed. PMID- 14530950 TI - Recurrent facial paralysis with plasma hypertriglyceridemia and hyperlipoproteinemia: case report. PMID- 14530951 TI - Anesthetic management for congenital erythropoietic porphyria: report of a case. PMID- 14530952 TI - Anesthetic management for a patient with acute intermittent porphyria treated with heme arginate. PMID- 14530953 TI - Electrocardiographic ST segment elevation during dopamine infusion for massive bleeding. PMID- 14530954 TI - Propofol suppressed electromyographic fibrillation potentials in a patient with amyotrophic lateral sclerosis. PMID- 14530955 TI - A patient with a preoperative hemoglobin concentration of 1.8 g.dl1: how was the life-threatening anemia tolerated without any intensive care? PMID- 14530956 TI - Target-controlled propofol infusion using STELPUMP with epidural analgesia or intravenous fentanyl. PMID- 14530957 TI - From oncology pharmacy to pharmaceutical care: new contributions to multidisciplinary cancer care. AB - In recent years a paradigm shift towards a patient-focused rather than a disease focused approach occurred in many health care systems. The pharmacy profession experienced an accordant development. The traditional drug-oriented services expanded towards patient-oriented services. In oncology, pharmacists established central services for compounding of cytotoxic drugs and offered therapeutic drug monitoring for critical substances. Pharmaceutical care concepts are now being introduced to optimize individual drug therapy. Pharmaceutical care aims at improving safety and therapeutic outcomes and consequently, the patient's quality of life. These objectives imply a close relationship to supportive care. To achieve this, a multidisciplinary approach seems to be beneficial. PMID- 14530958 TI - Batch and continuous cultures of Mannheimia succiniciproducens MBEL55E for the production of succinic acid from whey and corn steep liquor. AB - Mannheimia succiniciproducens MBEL55E isolated from bovine rumen is able to produce a large amount of succinic acid in a medium containing glucose, peptone, and yeast extract. In order to reduce the cost of the medium, whey and corn steep liquor (CSL) were used as substrates for the production of succinic acid by M. succiniciproducens MBEL55E. Anaerobic batch cultures of M. succiniciproducens MBEL55E in a whey-based medium containing CSL resulted in the production of succinic acid with a yield of 71% and productivity of 1.18 g/l/h, which are similar to those obtained in a whey-based medium containing yeast extract (72% and 1.21 g/l/h). Anaerobic continuous culture of M. succiniciproducens MBEL55E in a whey-based medium containing CSL resulted in a succinic acid yield of 69% and a succinic acid productivity as high as 3.90 g/l/h. These results show that succinic acid can be produced efficiently and economically by M. succiniciproducens MBEL55E from whey and CSL. PMID- 14530959 TI - Seasonality in daily body mass variation in a hoarding boreal passerine. AB - We studied the body mass variation from autumn to winter, in a free-living population of willow tits ( Parus montanus), a food-hoarding passerine living year-round in boreal forests. Our aim was to find out whether this population exhibits 'winter fattening' as part of the annual body mass cycle. 'True winter fattening' is considered to be a strategic response to winter conditions. The strategy includes an increase in both the morning mass and the daily mass increase, as winter approaches. A multivariate approach was used to find which predictors (year, date, age, sex, body size, temperature and snow depth) explained the mass variation in birds measured twice per day. Morning mass variation was explained by sex, age, wing length and snow depth. Independently, date explained morning mass variation only in adult males. None of the predictors explained the variation observed in daily mass increase in any age or sex class. Therefore, we failed to detect winter fattening in our study population of willow tits. Response to increasing night length is not due to higher absolute intake, but to higher energy acquisition rate and decreased night-time energy consumption. The results suggest that willow tits at high latitudes manage increasing energy demands on a short-term basis and respond flexibly to changing conditions by adjusting foraging efficiency and especially night-time energy expenditure. PMID- 14530960 TI - Carbon availability controls the growth of detritivores (Lumbricidae) and their effect on nitrogen mineralization. AB - Activity of soil decomposer microorganisms is generally limited by carbon availability, but factors controlling saprophagous soil animals remain largely unknown. In contrast to microorganisms, animals are unable to exploit mineral nutrient pools. Therefore, it has been suggested that soil animals, and earthworms in particular, are limited by the availability of nitrogen. In contrast to this view, a strong increase in density and biomass of endogeic earthworms in response to labile organic carbon addition has been documented in field experiments. The hypothesis that the growth of endogeic earthworms is primarily limited by carbon availability was tested in a laboratory experiment lasting for 10 weeks. In addition, it was investigated whether the effects of earthworms on microbial activity and nutrient mineralization depend on the availability of carbon resources. We manipulated food availability to the endogeic earthworm species Octolasion tyrtaeum by using two soils with different organic matter content, providing access to different amounts of soil, and adding labile organic carbon (glucose) enriched in (13)C. Glucose addition strongly increased the growth of O. tyrtaeum. From 8 to 17% of the total C in earthworm tissue was assimilated from the glucose added. Soil microbial biomass was not strongly affected by the addition of glucose, though basal respiration was significantly increased and up to 50% of the carbon added as glucose was incorporated into soil organic matter. The impact of earthworms on the mineralization and leaching of nitrogen depended on C availability. As expected, in C-limited soil, the presence of earthworms strongly increased nitrogen leaching. However, when C availability was increased by the addition of glucose, this pattern was reversed, i.e. the presence of O. tyrtaeum decreased nitrogen leaching and its availability to soil microflora. We conclude that irrespective of the total carbon content of soils, O. tyrtaeum was primarily limited by carbon, and that increased carbon availability allowed earthworms to be more effective in mobilizing N. The presence of earthworms increases C limitation of soil microorganisms, due to increased availability of N and P in earthworm casts or a direct depletion of easily available carbon resources by earthworms. PMID- 14530961 TI - Stable isotope ecology in the Ituri Forest. AB - The Ituri Forest, Democratic Republic of Congo (formerly Zaire) is an example of a closed canopy forest showing extreme depletion in (13)C. delta(13)C values for plants from the canopy top, from gaps in the canopy, and from the subcanopy average -29.0+/-1.7 per thousand, -30.4+/-0.9 per thousand, and -34.0+/-1.5 per thousand, respectively. The delta(13)C of forest mammals show these differences, with the subcanopy browsers (okapi, dwarf antelope) having delta(13)C values for tooth enamel much more negative than subcanopy frugivores who derive their food from the canopy top, and from folivores and omnivores living in gap or clearing areas. Nitrogen isotopes in plants from this ecosystem have an average delta(15)N value of 5.4+/-1.8 per thousand and do not show significant differences at the 95% confidence interval between plants from the canopy top, from gaps in the canopy, and from the subcanopy. The delta(18)O(SMOW) values of surface waters in the study area are between -2.0 and -2.7. The delta(18)O(PDB) for tooth enamel ranged from -3 to +7 per thousand. PMID- 14530962 TI - Larval cannibalism, time constraints, and adult fitness in caddisflies that inhabit temporary wetlands. AB - The fitness of non-feeding adult insects depends on energy accumulated during the larval stage. Larvae of the caddisfly Asynarchus nigriculus primarily feed on plant detritus, but supplement their diet with animal material obtained through cannibalism. Habitat drying constrains development in many populations of this species, and we hypothesized that cannibalism should accelerate development to facilitate timely metamorphosis. We manipulated larval diets in a field experiment by supplementing detritus with animal material, and in a laboratory experiment by varying animal material and detritus quality (conditioned vs unconditioned). We measured the effects of dietary manipulation on larval and pupal growth and development, the timing of metamorphosis, and adult fitness correlates. The results of the laboratory experiment suggest that this species can metamorphose with a detritus-only diet, but development is extremely protracted. In the field experiment, individuals with animal material in their diet had higher larval survival, shorter larval and pupal development times, and earlier emergence dates (7-10 days), than those without a supplement. This delay in emergence should have important effects on survival in natural populations where the difference between desiccation and successful emergence can be only a few days. Dietary supplementation also affected adult body mass (30-40% increase), female fecundity (30% more eggs), and proportional allocation to different adult body parts. Our results are consistent with recent growth development models that predict coupled (earlier emergence and larger adults) rather than tradeoff responses (earlier emergence and smaller adults) to pre threshold manipulation of larval diets. Many detritivorous aquatic insects supplement their diets with animal material, and our data provide evidence that this supplementation can have strong effects on fitness. This type of dietary supplementation should be especially important for taxa that do not feed as adults, and in temporary habitats that impose time constraints on larval development. PMID- 14530963 TI - Transcriptional regulation of the human LINE-1 retrotransposon L1.2B. AB - Although LINE-1 (L1) sequences constitute the most important family of retrotransposons in the human genome, their transcriptional regulation is poorly understood. Specifically, their unusual internal promoter is incompletely characterized. Current promoter prediction programs fail to identify the promoter in the 5'UTR of the active LINE-1 element L1.2B. Experimental investigation of this promoter using reporter gene assays in various human and murine cell types confirmed that the promoter consists of two segments, and demonstrated that the distal portion is essential for cell-type-independent activity. No differences in promoter activity were found between normal and transformed cells. The complete promoter was shown to possess approximately 20% of the activity of the strong early promoter of cytomegalovirus, and to be capable of directing the expression of levels of p53 sufficient to kill normal and transformed human cells. Thus, active LINE-1 elements contain highly active promoters capable of driving cell type-independent expression, which are of potential use in mammalian expression constructs. In vitro methylation of the promoter at HpaII sites decreased its activity independently of cell type, but this repression was alleviated in MBD2-/ cells. Surprisingly, mutation of specific HpaII sites was also found to reduce promoter activity. Thus, efficient repression of the L1.2B promoter by DNA methylation may involve MBD2 binding, but at least one HpaII site also appears to be involved specifically in transcriptional activation. Since neither promoter activity nor the efficiency of repression by methylation differed between normal and tumor cells, the re-activation of LINE-1 sequences observed in tumor cells is probably caused by hypomethylation of the promoter. PMID- 14530965 TI - Schizogonic stages of Haemoproteus from Wenyon's Baghdad sparrows are also found in Passer domesticus biblicus in Israel. AB - Schizont bodies reminiscent of those described by Wenyon from Baghdad sparrows were found in the liver and lungs of an Israeli house sparrow ( Passer domesticus biblicusHartert 1904) infected with Haemoproteus passeris Kruse, 1890. All observed schizonts were composed of packed assemblages of walled compartments, each holding a differentiating schizogonic body. The schizogonic bodies in the various compartments demonstrated sequential stages in the differentiation process from a compact multinucleate cytoplasmic mass to massive formation of multiple merozoites. Young non-differentiated schizont assemblages reached 0.2 x 0.25 mm in size and the fully differentiated ones, containing merozoites, were double or triple that. Among species of Haemopterus, division to compartmented and single-plasmodium schizonts could not be associated with any recognized generic or infrageneric division. Moreover, in some species of Haemoproteus, both types of schizogony co-existed. PMID- 14530967 TI - A rodent model for pulmonary paragonimiasis. AB - This study reports a model of pulmonary paragonimiasis in outbred Wistar rats using the Indian strain of Paragonimus heterotremus isolated from the fresh water crab Barytelphusa lugubris collected from Arunachal Pradesh, India. Feeding metacercariae to the rats led to encapsulated adult worms in the lungs and pleural cavity. Some flukes migrated to the skeletal muscles of the hosts where they remained underdeveloped as immature flukes. PMID- 14530966 TI - Targeting enzymes involved in spermidine metabolism of parasitic protozoa--a possible new strategy for anti-parasitic treatment. AB - Sequencing data obtained from the Plasmodium, Anopheles gambiae and human genome projects provide a new basis for drug and vaccine development. One of the most characteristic features in the process of drug development against parasitic protozoa is target identification in a biological pathway. The next step must be a structure-based rational drug design if the target is not only present in the parasite. In mouse models of malaria, such drugs should be tested for efficacy of the new therapies. Here, we present data that pinpoint the existence of two enzymes of the polyamine pathway involved in spermidine metabolism in P. falciparum, i.e. deoxyhypusine synthase (DHS; EC 1.1.1.249) and homospermidine synthase (HSS; EC 2.5.1.45). Recent data obtained from the malaria genome databases showed that at least a putative gene encoding DHS is present in the parasite. Sequencing data from the P. falciparum genome project prove that the eukaryotic initiation factor eIF5A (the substrate for DHS) exists in P. falciparum. Here, we present the amino acid sequence of eIF5A from P. vivax, which causes tertiary malaria. EIF5A from P. vivax shows 82% nucleic acid and 97% amino acid identity to its homologue from P. falciparum. GC/MS data and inhibitor studies with agmatine prove that the triamine homospermidine occurs in the parasite. These data suggest a separate locus encoding HSS in P. falciparum. The hss gene recruits from the dhs gene in eukaryotes. Here, we present genomic DNA fragments obtained by amplification with primers of a conserved region (amino acid positions 550-1,043) between the putative P. falciparum DHS gene ( dhs) and the HSS gene ( hss) from the plant Senecio vulgaris (Asteraceae). The amplification product from different P. falciparum strains reveals differences in sequence identity, compared with the putative dhs gene from P. falciparum strain 3D7. Expression of the full-length clone and determination of HSS-specific activity will finally prove whether a separate region encoding HSS exists. PMID- 14530968 TI - Population-S benzimidazole- and tetrahydropyrimidine-resistant small strongyles in a pony herd in Kentucky (1977-1999): effects of anthelmintic treatment on the parasites as determined in critical tests. AB - Population-S small strongyles have been studied since 1974 in central Kentucky in a closed Shetland pony breeding herd. The ponies were treated approximately every 8 weeks with cambendazole (1974-1978), oxibendazole (OBZ) (1978-1992), or pyrantel pamoate (PRT) (1992-1999). Small strongyles in the ponies have shown resistance to these compounds in field and critical tests. One purpose of this presentation was to compare different parameters for determination of effects on the small strongyle species in ponies after treatment, mainly with OBZ or PRT, from data in critical tests (n=112). Also, the objective was to report on relative changes in the composition of species of small strongyles during the period 1977 through 1999. The following entities were compared to evaluate the effect of OBZ- or PRT-treatment on the small strongyles: (1) numbers of specimens with eggs in utero--there were less gravid worms passed in the feces than recovered at necropsy for OBZ but the numbers of gravid worms were similar in both categories for PRT, (2) pre- and posttreatment counts of eggs per gram of feces (EPGs) and larvae per gram of feces (LPGs)--the reductions were greater for the counts of EPGs than LPGs for OBZ but not for PRT, and (3) pre- and posttreatment counts of EPGs versus % of worms removed--reductions of the former were greater than the latter for both compounds. As shown from data in this study, reduction of EPG counts post treatment indicated much greater drug activity than was actually demonstrated by removal of worms. One evident factor was the value of doing critical tests to verify posttreatment counts of EPGs as indicators of anthelmintic activity. Twenty-eight species of small strongyles were found. For the 20 most prevalent species in the study, two decreased, five remained unchanged, three increased and then became stationary, five increased but then decreased, and five increased progressively. Numbers of small strongyles were highest in 1987 and 1999. PMID- 14530964 TI - The Arabidopsis KAKTUS gene encodes a HECT protein and controls the number of endoreduplication cycles. AB - In animals and plants, many cell types switch from mitotic cycles to endoreduplication cycles during differentiation. Little is known about the way in which the number of endoreduplication cycles is controlled in such endopolyploid cells. In this study we have characterized at the molecular level three mutations in the Arabidopsis gene KAKTUS ( KAK), which were previously shown specifically to repress endoreduplication in trichomes. We show that KAK is also involved in the regulation of the number of endoreduplication cycles in various organs that are devoid of trichomes. KAK encodes a protein with sequence similarity to HECT domain proteins. As this class of proteins is known to be involved in ubiquitin mediated protein degradation, our finding suggests that the number of endoreduplication cycles that occur in several cell types is controlled by this pathway. The KAK gene defines a monophylogenetic subgroup of HECT proteins that also contain Armadillo-like repeats. PMID- 14530969 TI - Silica treatment increases the susceptibility of the Cabo Frio strain of Biomphalaria tenagophila to Schistosoma mansoni infection but does not alter the natural resistance of the Taim strain. AB - The present work demonstrates that silica treatment represents a suitable in vivo method to evaluate the role of host phagocytic hemocytes in the mechanisms of resistance to parasitic infection. Silica inoculation into Biomphalaria tenagophila snail induced a significant reduction in the circulating hemolymph granulocytes in both strains tested (Taim and Cabo Frio). The granulocyte reduction was accompanied by a significant increase in the number of circulating dead cells. In B. tenagophila Cabo Frio, silica treatment enhanced snail susceptibility to Schistosoma mansoni, shortening the intramolluskan phase of the parasite and increasing the number of sporocysts and cercariae produced. In B. tenagophila Taim, the same treatment did not abrogate natural resistance to S. mansoni reported for this snail strain. These in vivo results demonstrate that macrophage-like granulocytes are involved in the mechanism of S. mansoni sporocyst destruction in Cabo Frio snails and suggest that another, different mechanism may be responsible to the natural resistance of B. tenagophila Taim. PMID- 14530970 TI - The origin of Sarcoptes scabiei in wombats. AB - In 2002, Skerratt et al. phylogenetically analysed sequence data for several haplotypes of the parasitic mite Sarcoptes scabiei from wombat, human and dog hosts in Australia, to test scenarios concerning the origin and diversification of the scabies infections in wombats. Here we note that their substantive conclusions can be called into question by the choice of model used in their phylogenetic analysis, the lack of a root for their phylogenetic trees, and their interpretation of the evolutionary scenario. PMID- 14530971 TI - Detection of Echinococcus multilocularis antigens in faeces by ELISA. AB - Faecal samples deriving from 391 animals belonging to nine species (polecats, badgers, martens, weasels, rats, dogs, cats, red foxes, raccoon-dogs) were examined by capture ELISA for the presence of the Echinococcus multilocularis coproantigen. The main claim of our studies is the reliable detection of E. multilocularis coproantigens, mainly in the faeces of foxes, dogs and cats. For the first time in coproantigen detection we used a "double-sandwich" ELISA. The main advantage of this method is the higher specificity and better differentiation of positive and negative faecal samples, in comparison with sandwich ELISA. The overall specificity of double-sandwich ELISA was 95.1% with only 16 of 327 E. multilocularis-free animals giving false-positive results. The E. multilocularis coproantigen was detected by double-sandwich ELISA in 37.5% of examined red foxes and in 8.0% of examined raccoon-dogs, compared with a prevalence of just 29.8% in red foxes and 8.0% in raccoon-dogs, as determined by parasitological techniques. PMID- 14530972 TI - The morphological features of the rete testis of the ostrich (Struthio camelus). AB - The epithelium of the rete testis of the ostrich is simple cuboidal to columnar in nature, unlike the more flattened epithelium encountered in other non passerine birds. A solitary cilium projects into the ductal lumen. A unique lateral cell membrane modification, similar, in some respects, to a hemi desmosome, occurs frequently along the length of this membrane. The lateral cell membrane is complexly folded, and the cell contains an abundance of intermediate filaments, particularly in the vicinity of the nucleus and the internal surface of the basal cell membrane. The Golgi complex and endoplasmic reticulum are moderately developed. Another unique feature of the cell is the presence of a solitary, large, heterogeneous lipid body/droplet situated in the immediate supranuclear region of the cell. Overall, the cell structure conveys the impression of a metabolically active cell, which has the capability of transporting material from the duct lumen to the basal part of the cell, and of protein synthesis, apparently much in excess of what is necessary for cellular maintenance and sustenance. PMID- 14530973 TI - SLC11 family of H+-coupled metal-ion transporters NRAMP1 and DMT1. AB - NRAMP1 (natural resistance-associated macrophage protein-1) and DMT1 (divalent metal-ion transporter-1) make up the SLC11 gene family of metal-ion transporters that are energized by the H(+) electrochemical gradient. Long known to confer resistance to bacterial infection, NRAMP1 functions at the phagolysosomal membrane of macrophages and neutrophils. NRAMP1 most likely contributes to macrophage antimicrobial function by extruding essential metal ions (including Mn(2+)) from the phagolysosome via H(+)/metal-ion cotransport. An alternative hypothesis in the literature proposes that NRAMP1 concentrate Fe(2+) within the phagolysosome by means of H(+)/Fe(2+) antiport, resulting in the generation of toxic free radicals. DMT1 is expressed widely and accepts as substrates a broad range of transition metal ions, among which Fe(2+) is transported with high affinity ( K(0.5) approximately 2 microM). DMT1 accounts both for the intestinal absorption of free Fe(2+) and for transferrin-associated endosomal Fe(2+) transport in erythroid precursors and many other cell types. DMT1 is up-regulated dramatically in the intestine by dietary iron restriction and, despite high serum iron levels, is not appropriately down-regulated in hereditary hemochromatosis. PMID- 14530976 TI - An MR-compatible device for the in situ assessment of isometric contractile performance of mouse hind-limb ankle flexors. AB - The goal of the present study was to develop and evaluate an isometric dynamometer for measuring mouse ankle flexor torque after electric stimulation of the nerve. The dynamometer was to be used within an magnetic resonance (MR) apparatus and should require minimal surgical intervention. To quantify the effect of the magnetic field on contractile parameters, measurements were performed both outside and inside the MR apparatus. The effect of magnetic field gradient switching that accompanies rapid MR scanning was tested also. The set-up required no surgical intervention except for chronic implantation of an electrode. The dynamometer has a high mechanical frequency response (270 Hz). Measured muscle strengths were identical outside and inside the MR scanner. However, during fast magnetic field gradient switching, the variability increased and the measured strength decreased slightly (7%). The noise level of the dynamometer (0.02-0.03 N.mm) was low compared with the strength of the dorsal flexors (2 N.mm). Fast gradient switching increased the noise level (0.07 N.mm). The dynamometer had no observable adverse effects on the quality of the MR images of the mouse hind limb. We conclude that the dynamometer enables accurate measurements of mechanical muscle performance during exercise protocols within an MR apparatus under physiological conditions. PMID- 14530974 TI - The glutamate/neutral amino acid transporter family SLC1: molecular, physiological and pharmacological aspects. AB - The solute carrier family 1 (SLC1) includes five high-affinity glutamate transporters, EAAC1, GLT-1, GLAST, EAAT4 and EAAT5 (SLC1A1, SLC1A2, SLC1A3, SLC1A6, and SLC1A7, respectively) as well as the two neutral amino acid transporters, ASCT1 and ASCT2 (SLC1A4 and ALC1A5, respectively). Although each of these transporters have similar predicted structures, they exhibit distinct functional properties which are variations of a common transport mechanism. The high-affinity glutamate transporters mediate transport of l-Glu, l-Asp and d-Asp, accompanied by the cotransport of 3 Na(+) and 1 H(+), and the countertransport of 1 K(+), whereas ASC transporters mediate Na(+)-dependent exchange of small neutral amino acids such as Ala, Ser, Cys and Thr. The unique coupling of the glutamate transporters allows uphill transport of glutamate into cells against a concentration gradient. This feature plays a crucial role in protecting neurons against glutamate excitotoxicity in the central nervous system. During pathological conditions, such as brain ischemia (e.g. after a stroke), however, glutamate exit can occur due to "reversed glutamate transport", which is caused by a reversal of the electrochemical gradients of the coupling ions. Selective inhibition of the neuronal glutamate transporter EAAC1 (SLC1A1) may be of therapeutic interest to block glutamate release from neurons during ischemia. On the other hand, upregulation of the glial glutamate transporter GLT1 (SLC1A2) may help protect motor neurons in patients with amyotrophic lateral sclerosis (ALS), since loss of function of GLT1 has been associated with the pathogenesis of certain forms of ALS. PMID- 14530975 TI - Role of nitric oxide in thermoregulation during septic shock: involvement of vasopressin. AB - We tested the hypothesis that the nitric oxide (NO) pathway in the central nervous system (CNS) plays a role in hypothermia, as well as in the febrile response during experimental septic shock, by regulating vasopressin (AVP) release. Experiments were performed on male Wistar rats treated with NG-nitro-L arginine methyl ester (L-NAME), a non-selective NO synthase (NOS) inhibitor, injected intracerebroventricularly (250 microg/1 microl) 30 min before lipopolysaccharide (LPS) 1.5 mg/kg i.v. injection. One hour after LPS administration we observed a significant drop in body temperature (hypothermic response), followed by a temperature increase after the second hour (febrile response), which remained until the end of the experiment. Increased plasmatic AVP levels were concomitantly observed during hypothermia, nearly returning to basal levels during the febrile phase. When L-NAME was administered with LPS, plasmatic AVP concentrations remained high throughout the experiment, hypothermia was accentuated and the febrile response was abolished. Additionally, pre treatment with beta-mercapto-beta,beta-cyclopentamethylenepropionyl1, O-Et-Tyr2, Val4, Arg8-vasopressin, an AVP V1 receptor blocker (10 microg/kg) administered i.v., reduced hypothermia and exacerbated the febrile response to endotoxin. In conclusion, our data indicate that the central NO pathway plays an inhibitory role in AVP release during experimental septic shock, which seems to be critical for the thermoregulation during this pathophysiological state. PMID- 14530977 TI - Ser16-, but not Thr17-phosphorylation of phospholamban influences frequency dependent force generation in human myocardium. AB - Beta-adrenoceptor/cAMP-dependent Ser16-phosphorylation as well as Ca(2+) dependent Thr17-phosphorylation of phospholamban (PLB) influences SERCA 2a activity and thus myocardial contractility. To determine the cross-signaling between Ca2+ and cAMP pathways, the phosphorylation of Ser16-PLB and Thr17-PLB was studied at increasing stimulation frequencies as well as in the presence of beta-adrenergic stimulation in isolated ventricular trabeculae from failing (dilative cardiomyopathy, DCM, heart transplants, n=9) and non-failing human myocardium (donor hearts, NF, n=9). In addition, we measured the intracellular Ca(2+)-transient (fura-2) at increasing stimulation frequencies (0.5-3.0 Hz). Protein expression of SERCA 2a and phospholamban was similar in DCM and NF. In DCM, diastolic [Ca2+]i was increased and systolic [Ca2+]i as well as Ser16 PLB phosphorylation were decreased as compared to NF at 0.5 Hz. The positive force frequency relationship in human non-failing myocardium was accompanied by a frequency-dependent increase in Ser16-PLB, but not Thr17-PLB phosphorylation. In DCM, Ser16-PLB as well as Thr17-PLB phosphorylation were not altered at higher stimulation frequencies. After application of isoprenaline (1 microM), a profound increase in Ser16-PLB phosphorylation was accompanied by a small increase in Thr17-PLB phosphorylation, only in NF. The frequency-dependent phosphorylation of Ser16-PLB may favor an increase in Ca2+ transient and force generation in humans. Cross talk signaling of Ser16/Thr17-PLB phosphorylation after beta-adrenergic stimulation exists in non-failing, but not in failing human myocardium. The Ca(2+)-dependent CaM-kinase activity may be altered in human heart failure. PMID- 14530979 TI - Gear, inertial work and road slopes as determinants of biomechanics in cycling. AB - In cycling the gear determines the distance travelled and the mean applied force at each leg thrust. According to Padilla et al. (J Appl Physiol 89:1522-1527, 2000), an elite cyclist was able to cycle for an hour at 14.6 m.s(-1 developing 510 W at a pedal frequency of 101 rpm. Thus, the opposing force was 34 N (=500/14.6), whereas the mean force, developed by the leg muscles, was 144.1 N. It can be calculated that in the same subject cycling on a 20% slope at the same pedal frequency, the velocity would be reduced by about 5 times, i.e. to 2.9 m.s( 1) because of a fivefold increase of the opposing force. In reality, the increase of mean force developed by leg muscles is even larger, because of the fall of the cadence to 60 rpm. In general, during mountain ascents cyclists develop high forces at low cadences that are likely to be more economical; in contrast, on flat ground, they increase the pedalling rates because their aerodynamic posture does not allow high force production. The intermittent pattern of muscular force application generates speed changes that become more evident at great inclines and low cadences. It can be shown that inertial work is appreciable in cycling, increasing with the incline of the road and decreasing with the cadence. However, inertial work does not seem to affect efficiency. Differences in physiologic potential make differences in performance more evident in time trials where the mean incline of the road is not negligible. Cyclists with low body size have an advantageous force versus mass ratio in high mountain ascents. PMID- 14530978 TI - Experimental models to study microcirculatory dysfunction in muscle ischemia reperfusion and osteomyocutaneous flap transfer. AB - BACKGROUND: During the past decade, experimental studies have provided convincing evidence that microcirculatory dysfunction plays a pivotal role in the manifestation of tissue injury in ischemia-reperfusion and osteomyocutaneous flap transfer. The study of the mechanisms of injury, however, requires sophisticated experimental in vivo models. With the use of microsurgical techniques, osteomyocutaneous flap transfer can successfully be performed in rat hind limbs, allowing in vivo fluorescent microscopic analysis of post-ischemic microcirculatory dysfunction in all tissues involved, including periosteum, striated muscle, subcutis and skin. The drawback of this "acute" model is that the period of analysis is restricted to a few hours only. METHOD: To overcome this limitation, the "chronic" dorsal skinfold chamber preparation, containing striated muscle and subcutis, can be used. This model allows one to study microcirculatory dysfunction after both tourniquet-induced and pressure-induced ischemia-reperfusion-induced tissue injury over a period of up to 3 weeks. RESULTS: With the use of these models, recent investigations have demonstrated that ischemia-reperfusion and osteomyocutaneous flap transfer are associated with capillary perfusion failure (no-reflow), mediated by intravascular hemoconcentration, endothelial swelling and endothelin (ET)-1-mediated microvascular constriction. In addition, post-ischemic reperfusion provokes an inflammatory response (reflow paradox) in post-capillary venules, which is characterized by beta2-integrin-mediated and intercellular adhesion molecule (ICAM)-1-mediated leukocyte adhesion and vascular hyperpermeability, which results in interstitial edema formation. Treatment studies have produced evidence that isovolemic hemodilution and heat shock protein induction are successful in ameliorating capillary no-reflow, while blockade of adhesion molecules, inactivation of oxygen radicals and, also, induction of heat shock proteins, are capable of reducing the post-ischemic inflammatory response. CONCLUSION: These experimental results not only demonstrate the importance of the use of advanced in vivo methods to delineate pathophysiological mechanisms in complex disease models, but may also provide a basis for potential prospective randomized trials to test the benefit for the patient in the daily clinical routine. PMID- 14530980 TI - Limiting factors to oxygen transport on Mount Everest 30 years after: a critique of Paolo Cerretelli's contribution to the study of altitude physiology. AB - In 1976, Paolo Cerretelli published an article entitled "Limiting factors to oxygen transport on Mount Everest" in the Journal of Applied Physiology. The paper demonstrated the role of cardiovascular oxygen transport in limiting maximal oxygen consumption (VO2max). In agreement with the predominant view of VO2max limitation at that time, however, its results were taken to mean that cardiovascular oxygen transport does not limit VO2max at altitude. So it was argued that the limiting factor could be in the periphery, and muscle blood flow was proposed as a possible candidate. Despite this suggestion, the conclusion generated a series of papers on muscle structural characteristics. These experiments demonstrated a loss of muscle oxidative capacity in chronic hypoxia, and thus provided an unambiguous refutation of the then widespread hypothesis that an increased muscle oxidative capacity is needed at altitude to compensate for the lack of oxygen. This analysis is followed by a short account of Cerretelli's more recent work, with a special attention to the subject of the so called "lactate paradox". PMID- 14530981 TI - Cardioventilatory changes induced by mentally imaged rowing. AB - Mentally imaged but unexecuted physical activity has been reported to induce a cardiorespiratory change. In order to test whether the previous experience of the performed exercise was a prerequisite to observe these changes, ventilation and heart rate were recorded during mental imagination of a rowing race in four groups of volunteers: 12 competitive rowers, 10 non-rower athletes, 12 students (22-30 years old) and 12 senior subjects (50-60 years old). Recordings were performed at rest, during the viewing of a rowing race and during mental imagination of this race. Analysis of variance revealed significant condition effect for all cardiorespiratory variables. All subjects increased their breathing rate (mean increase: 16 breaths.min(-1) in rowers, 8 breaths.min(-1) in athletes, 8 breaths.min(-1) in students, and 6 breaths.min(-1) in seniors), 29 decreased their tidal volume (mean decrease: 100 ml in rowers, 102 ml in athletes, 120 ml in students and 26 ml in seniors), with an increase in the resulting ventilation in 38 subjects (mean increase: 14 l.min(-1) in rowers, 3.6 l.min(-1) in athletes, 2.8 l.min(-1) in students, 2.6 l.min(-1) in seniors). Heart rate was increased in 34 subjects (mean increase: 12 beats.min(-1) in rowers, 5 beats.min(-1) in athletes, 6 beats.min(-1) in students and 5 beats.min( 1) in seniors). The number of subjects who exhibited changes was evenly distributed among the four groups. However, mean values of the changes were higher in rowers than in the three other groups, mainly due to three rowers who exhibited extremely large increases in cardioventilatory variables. Analysis of variance showed no significant group effect for heart rate and breathing rate. These results suggest that rowing experience may not be necessary for changes in heart rate and ventilation to be elicited by mentally imagining a rowing race. PMID- 14530982 TI - Effects of power training on mechanical efficiency in jumping. AB - The present study investigates the effects of power training on mechanical efficiency (ME) in jumping. Twenty-three subjects, including ten controls, volunteered for the study. The experimental group trained twice a week for 15 weeks performing various jumping exercises such as drop jumps, hurdle jumps, hopping and bouncing. In the maximal jumping test, the take-off velocity increased from 2.56 (0.24) m.s(-1) to 2.77 (0.18) m.s(-1) ( P<0.05). In the submaximal jumping of 50% of the maximum, energy expenditure decreased from 660 (110) to 502 (68) J.kg(-1).min(-1) ( P<0.001) while, simultaneously, ME increased from 37.2 (8.4)% to 47.4 (8.2)% ( P<0.001). Some muscle enzyme activities of the gastrocnemius muscle increased during the training period: citrate synthase from 35 (8) to 39 (7) micromol.g(-1) dry mass.min(-1) ( P<0.05) and beta-hydroxyacyl CoA dehydrogenase from 21 (4) to 23 (5) micromol.g(-1) dry mass.min(-1) ( P<0.05), whereas no significant changes were observed in phosphofructokinase and lactate dehydrogenase. In the control group, no changes in ME or in enzyme activities were observed. In conclusion, the enhanced performance capability of 8% in maximal jumping as a result of power training was characterized by decreased energy expenditure of 24%. Thus, the increased neuromuscular performance, joint control strategy, and intermuscular coordination (primary factors), together with improved aerobic capacity (secondary factor), may result in reduced oxygen demands and increased ME. PMID- 14530984 TI - Confirmation of alleged falanga torture by bone scintigraphy-Case report. AB - Any objective persisting signs of previous torture would be very valuable in the late assessment of the individual claiming such abuse of human rights. We present the case of a 32-year-old man referred to our hospital for an opinion on alleged torture by the falanga method. Magnetic resonance imaging and bone scintigraphy were evaluated and compared as methods of confirming such torture. PMID- 14530983 TI - A case-control study of occupational exposures and systemic sclerosis. AB - OBJECTIVES: A case-control study was conducted in Verona, Italy, to assess the relationship between occupation, occupational exposures and systemic sclerosis (SSc). METHODS: Fifty-five cases (46 female and nine male) and 171 controls were recruited. Interviews provided work histories, including job titles, industry and likelihood of occupational exposure to silica, hand-arm vibration, organic solvents, and other chemicals. Odds ratios (ORs) and 95% confidence intervals (95% CI) were estimated. RESULTS: Female teachers (OR 3.4, 95% CI 1.2-10.1) and textile workers (OR 2.1, 95% CI 1.0-4.6) were at an increased risk of SSc. Compared with those never exposed, age-adjusted and gender-adjusted ORs were 2.3 (95% CI 1.0-5.4) among subjects exposed to organic solvents, 2.5 (95% CI 0.8-8.0) for exposure to selected chemicals, 1.7 (95% CI 0.4-7.6) for exposure to silica, and 1.5 (95% CI 0.5-4.8) for usage of vibrating tools. When data analysis was stratified according to gender, only men showed a significant increase in risk for exposure to solvents and selected chemicals. CONCLUSIONS: The findings of this study tend to support the role of organic solvents and certain chemicals in SSc causation. The association with teaching and working in the textile industry suggests that other exposures are involved in the aetiology of SSc among women. However, because of the small number of subjects, particularly in stratified analyses, chance cannot be ruled out as an explanation of some findings of this study. PMID- 14530985 TI - LC-MS determination of Taxus alkaloids in biological specimens. AB - A semi-quantitative LC-MS method was developed for the detection of the pseudo alkaloids of Taxus baccata (yew) from human body fluids and tissue samples. This method was used to examine the cause of death of a 43-year-old man who died several hours after he drank a decoction of taxus leaves. Autopsy and histology demonstrated early signs of myocardial hypoxia. Since investigation of the stomach content did not yield evidence of taxus ingestion, the taxus alkaloids were determined in blood, stomach content and tissue samples of the deceased by LC-MS. The samples were prepared by solid phase extraction on RP-18 columns. Chromatographic separation was achieved by HPLC on a RP-8 column, coupled to an ion trap mass spectrometer (Finnigan LCQ). An atmospheric pressure electrospray ionisation was performed. Spectra of the alkaloids were recorded in the single MS mode and in the MS-MS mode and compared with reference spectra obtained from an extract of yew leaves. In the stomach content, the kidneys, the liver and a heart blood sample of the deceased, alkaloids of Taxus baccata, predominantly taxine B and iso-taxine B, were identified. The semi-quantitative evaluation of the heart blood revealed a taxine concentration of 11 micro g taxine/g. As far as we know this is the first case in which a semi-quantitative analysis of taxine alkaloids has been performed. PMID- 14530986 TI - The signature of the Cestrum genome suggests an evolutionary response to the loss of (TTTAGGG)n telomeres. AB - The genus Cestrumin the Solanaceae family is unusual in lacking Arabidopsis-type telomeres (TTTAGGG)n, although short interstitial telomeric sequences (ITSs) occur scattered throughout the genome in both orientations. To isolate candidate telomeric sequences in Cestrum we assumed that some of the ITSs were residues of the original telomeres and that they may still be located in the vicinity of present-day telomeres. Three sequence types associated with ITSs were cloned and characterized; these were termed NA3G, BR23 and A/T-rich minisatellite. These high copy number sequences are dispersed across the genome and clustered at a number of chromosomal loci. Their association with ITSs, which can act as recombination hotspots, might indicate past recombination and chromosomal fusion events, processes that may have contributed to the large size of Cestrum chromosomes. The sequences are frequently arranged as NA3G-ITS-BR23 blocks embedded in an A/T-rich minisatellite array. The A/T-rich minisatellite is of particular interest because the consensus 5'-T(4-5)AGCAG-3' might be a derivative of "typical" eukaryotic telomeric sequence motifs. The sequence is abundant at the end of some chromosomes in C. parqui and is found not only in Cestrum but also in the closely related genera Sessea and Vestia, which also lack Arabidopsis type telomeric sequences. However, the sequence is absent from the Solanaceae genera investigated that are outside the group, including the closely related genus Streptosolen, which all have the Arabidopsis-type telomere. The data indicate that the A/T rich minisatellite might have evolved in response to the loss of Arabidopsis-type telomeres. PMID- 14530987 TI - Influence of cultured dermal fibroblasts on human melanoma cell proliferation, matrix metalloproteinase-2 (MMP-2) expression and invasion in vitro. AB - The dermis is the main site of melanoma invasion. Matrix metalloproteinases (MMPs), especially MMP-2, produced by melanoma or surrounding stromal cells, are essential for the destruction of dermal extracellular matrix. Here, we examined how dermal fibroblasts influenced proliferation, MMP-2 secretion and invasion of human melanoma cell lines in vitro. Human melanoma cell lines M3 Da and M1Dor were cocultured with dermal fibroblasts under non-contact and contact conditions in order to assess both soluble and insoluble factors, respectively. Zymographic analysis showed that the levels of MMP-2 and TIMP-2 in melanoma cells were not altered in non-contact cocultures when compared with those in individual cultures. However, in contact cocultures, the expression of MMP-2 in membrane extracts was enhanced. Under our coculture conditions, dermal fibroblasts failed to upregulate melanoma cell invasion through a three-dimensional type I collagen matrix. Since stromal and cancer cell contacts have been shown to occur after disruption of the extracellular matrix, we hypothesized that fibroblasts may influence melanoma cell invasion after the beginning of tumor progression through the dermis. PMID- 14530988 TI - Molecular evidence that halo in Sutton's naevus is not vitiligo. AB - Both halo naevus and vitiligo are acquired leucodermas of unknown aetiology. To date a significant contribution of oxidative stress through accumulation of hydrogen peroxide (H2O2) has been documented in the pathomechanism of vitiligo but not in halo naevus. Both epidermal pterin-4a-carbinolamine dehydratase (PCD) and catalase are sensitive markers to follow H2O2 concentration-dependent deactivation of these proteins. In situ protein expression of PCD and catalase was examined in full-skin biopsies from skin phototype-matched controls (n=5), untreated and treated vitiligo patients (n=5) and patients with untreated halo naevus in association with vitiligo (n=3). Vitiligo was treated with pseudocatalase (PC-KUS) only. Catalase levels were determined in epidermal suction blister extracts using fast protein liquid chromatography (FPLC). In addition, epidermal H2O2 levels were followed in vivo by Fourier-transform Raman spectroscopy. The results of this study ruled out a contribution of H2O2 in the millimolar range in the depigmentation process of halo naevus as previously documented in vitiligo. Therefore, it can be concluded that both leucodermas exercise distinct concentration-dependent H2O2 signalling in their pathomechanisms. PMID- 14530989 TI - Effects of induced hypothermia after soft-tissue injury. AB - BACKGROUND: Hypothermia is occasionally employed to reduce the metabolic rate and to protect the brain in patients undergoing surgery, but it is controversial whether hypothermia (HT) is beneficial or harmful in trauma victims with soft tissue injuries. For this purpose, we studied the acute effects of hypothermia induced after infliction of a standardized soft-tissue injury. METHODS: After a standardized high-energy gunshot wound to the right hind leg of 14 anesthetized piglets, the animals were randomized to normothermia or HT (30 degrees C) induced with a HT bed. The cardiovascular and hematological status was monitored for 6 h after the injury. RESULTS: The heart rate, mean arterial pressure, neutrophil count, and plasma adrenaline level were significantly lower in the HT pigs than in the controls (p<0.05). The arterial oxygen tension was significantly elevated in the HT group. Cardiac index and oxygen delivery decreased slightly in both groups, but no difference developed between the groups in these parameters. Serum potassium increased significantly in the controls (p<0.001). CONCLUSION: HT down to 30 degrees C following high-energy penetrating soft-tissue injury had a modest effect on the hemodynamics and oxygen delivery. However, evidence of 'stress' was reduced, and the pigs developed a progressive increase in their serum potassium concentration. PMID- 14530990 TI - Ultrasonographic evaluation at 6-month follow-up of plantar fasciitis after extracorporeal shock wave therapy. AB - INTRODUCTION: The aim of this study was to investigate the effect of extracorporeal shock wave therapy (ESWT) on the ultrasonographic appearance of chronically painful, proximal plantar fasciitis. MATERIALS AND METHODS: Twenty two patients with a unilateral proximal plantar fasciitis were prospectively enrolled after unsuccessful conservative treatment lasting 6 months. The contralateral plantar fascia was used as the control. ESWT (3x3000 shock waves/session of 0.2 mJ/mm2) was performed at weekly intervals. The thickness of the plantar fascia was measured ultrasonographically about 2 cm distal of the medial calcaneal tuberosity. Pain estimation on a visual analogue scale (VAS) and the comfortable walking time were recorded. No local anaesthesia was applied. Follow-up was done at 6, 12 and 24 weeks. RESULTS: Before ESWT, the plantar fasciitis side was ultrasonographically significantly thicker than the control side (p<0.05), whereas 6 months after ESWT, the thickness of the fascia was no longer significantly different. The decrease in thickness of the plantar fasciitis side was significant (p<0.05). Pain during activities of daily living decreased by 79% according to the VAS, and the comfortable walking time increased, both significantly (p<0.01). In patients with little pain (VAS<30), the thickness of the plantar fasciitis side was significantly less (p<0.01) compared with patients who still suffered more pain (VAS>30). CONCLUSION: After ESWT, the thickness of the plantar fascia in patients with plantar fasciitis decreased, pain and walking time improved (all significantly). PMID- 14530991 TI - Longitudinal pathologic study of the gastrocnemius muscle group in mdx mice. AB - Muscle necrosis in mdx mice is rare before the 2nd week of life, but becomes pronounced from weeks 2 to 6. There is no agreement on what happens after this age. Using sequential microscopic histology, immunohistochemistry and histomorphometry, we studied the evolution of muscle pathology throughout the mdx life span. Between 2 weeks and 3 months, multiple necrotic groups involving variable numbers of fibres were observed in the same section. During this period, most necrotic groups comprised more than 15 fibres. From 3 to 6 months, necrosis diminished progressively, involving isolated fibres or groups of two to five fibres, and most fibres had a centrally located nucleus. From 6 months onward there was progressive variation of fibre size and deposition of fibrillary collagens. Mast cell counts increased progressively until 6 months of age. Diminishing necrosis rather than increasing regeneration may explain the differences in muscle pathology observed between human DMD and mouse mdx. Mast cells play a secondary role in pathogenesis. PMID- 14530992 TI - Customized biofeedback therapy improves results in fecal incontinence. AB - BACKGROUND AND AIMS: Biofeedback therapy has been extensively used and accepted in fecal incontinence, but reports of its efficiency vary. We evaluated feedback therapy efficiency when (a) selecting the patient's subject of the therapy, and (b) customizing the therapy protocol used for each patient. PATIENTS AND METHODS: Fifty-three patients with fecal incontinence were selected for biofeedback training. The treatment program was customized for each patient depending on the underlying dysfunction, the patient's cooperative and learning attitude, and the patient's progress. Biofeedback efficiency was measured using clinical scores, subjective satisfaction of the patient, and manometry. RESULTS: Incontinent scores showed improvement in 66% of patients and good improvement in 11% and 15%, respectively, indicating an overall excellent effect of the therapy. Subjective satisfaction was strongly correlated with the previous incontinent scores. Comparison of manometry parameters before and after biofeedback therapy, including maximum anal resting, maximum anal squeeze pressure, and maximum duration of the squeeze, all showed significant differences. In addition, the sensory threshold significantly decreased after biofeedback therapy. Clinical improvements were maintained during the following 12 months. CONCLUSION: Biofeedback improves objective and subjective parameters of anorectal function. Selection of patients and customization of the therapy program increased biofeedback efficiency for the treatment of fecal incontinence. PMID- 14530994 TI - Surgery of rectal cancer: still not a science. PMID- 14530993 TI - Restorative resection of unprepared left-colon in gangrenous vs. viable sigmoid volvulus. AB - BACKGROUND AND AIMS: Emergency resection and primary anastomosis of unprepared left-colon is a controversial subject. Although this approach has been reported in several series, there is paucity of data on the relative safety of it in viable vs. gangrenous colon especially when the gut is unprepared. PATIENTS AND METHODS: Case records of 57 consecutive patients with acute sigmoid volvulus were reviewed; there were 27 with gangrenous colon (group G) and 30 with viable colon (group V). All of them had undergone emergency resection and primary anastomosis without on-table lavage or caecostomy. RESULTS: Group G had a lower mean haemoglobin value (8.4 vs. 9.7 g/dl) and higher incidence of circulatory shock on admission (26% vs. 7%) and required more blood transfusion (85% vs. 53%) than group V. Mean hospital stay (16 vs. 12 days), overall anastomotic leak (15% vs. 27%) and mortality (3.5% vs. 3%) did not differ significantly between the groups. However, the rate of wound infection in Group G was four times greater than that of group V. CONCLUSION: One-stage restorative resection without on-table lavage or caecostomy appears to be a promising alternative in the emergency management of acute sigmoid volvulus. Comparison of primary anastomosis in gangrenous vs. viable colon did not reveal any significant difference in hospital stay, rate of anastomotic leak or mortality. However, the risk of wound infection was more in patients with gangrenous sigmoid volvulus. PMID- 14530995 TI - Pelvic sidewall involvement in recurrent rectal cancer. AB - BACKGROUND AND AIMS: The lateral pelvic sidewall is an area not routinely dissected during standard operative procedures in surgery for rectal cancer in Western countries. This study analyzed data to evaluate the pattern of recurrence in rectal cancer with special emphasis on lateral tumor extension in a recently treated patient population. PATIENTS AND METHODS: In a multicenter retrospective study 123 patients were evaluated by our own CT-based three-dimensional datafile system and an extensive questionnaire. Patients had histological confirmation, clear bone destruction, a positive PET scan, and at least three minor criteria: progressive soft tissue mass, invasion of adjacent organs on follow-up CT or MRI, rising tumor markers, and typical appearance in cross-sectional imaging. Clinical or serological signs of inflammation were exclusion criteria. Initially 54% of the evaluated patients were N0, and the others were distributed evenly between N1 and N2; initial T stage was T1 in 2%, T2 in 24%, T3 in 60%, and T4 in 13%. RESULTS: . Recurrent tumors were situated mainly in the posterior part of the bony pelvis. The pelvic side wall was a rare site of recurrence and involved in fewer than 5%. When abdominoperineal resection was compared to low anterior resection as primary operation, there was a significant difference in extension of recurrent tumors in the inferior parts of the pelvis; no significant differences were found in superior or lateral parts of the pelvis. CONCLUSION: Because most tumor recurrences arise in the central pelvis, extending surgery to include dissecting the iliac vessels would probably offer only a moderate benefit, which must be balanced against potential side effects. PMID- 14530996 TI - L-type Ca2+ channel and Ca2+-permeable nonselective cation channel as a Ca2+ conducting pathway in myocytes isolated from the cricket lateral oviduct. AB - The Ca2+-conducting pathway of myocytes isolated from the cricket lateral oviduct was investigated by means of the whole-cell patch clamp technique. In voltage clamp configuration, two types of whole cell inward currents were identified. One was voltage-dependent, initially activated at -40 mV and reaching a maximum at 10 mV with the use of 140 mM Cs2+-aspartate in the patch pipette and normal saline in the bath solution. Replacement of the external Ca2+ with Ba2+ slowed the current decay. Increasing the external Ca2+ or Ba2+ concentration increased the amplitude of the inward current and the current-voltage (I-V) relationship was shifted as expected from a screening effect on negative surface charges. The inward current could be carried by Na+ in the absence of extracellular Ca2+. Current carried by Na+ (INa) was almost completely blocked by the dihydropyridine Ca2+ channel antagonist, nifedipine, suggesting that the INa is through voltage dependent L-type Ca2+ channels. The other inward current is voltage-independent and its I-V relationship was linear between -100 mV to 0 mV with a slight inward rectification at more hyperpolarizing membrane potentials when 140 mM Cs+ aspartate and 140 mM Na+-gluconate were used in the patch pipette and in the bath solution, respectively. A similar current was observed even when the external Na+ was replaced with an equimolar amount of K+ or Cs+, or 50 mM Ca2+ or Ba2+. When the osmolarity of the bath solution was reduced by removing mannitol from the bath solution, the inward current became larger at negative potentials. The I-V relationship for the current evoked by the hypotonic solution also showed a linear relationship between -100 mV to 0 mV. Bath application of Gd3+ (10 microM) decreased the inward current activated by membrane hyperpolarization. These results clearly indicate that the majority of current activated by a membrane hyperpolarization is through a stretch-activated Ca2+-permeable nonselective cation channel (NSCC). Here, for the first time, we have identified voltage dependent L-type Ca2+ channel and stretch-activated Ca2+-permeable NSCCs from enzymatically isolated muscle cells of the cricket using the whole-cell patch clamp recording technique. PMID- 14530998 TI - Microbubble ultrasound contrast in the assessment of hepatic artery patency following liver transplantation: role in reducing frequency of hepatic artery arteriography. AB - We prospectively evaluated the role of microbubble ultrasound contrast for detection of hepatic artery thrombosis following liver transplantation. The hepatic artery of adult liver transplant recipients with suspected thrombosis on surveillance Doppler ultrasound (US) were re-examined by a second observer. In patients with no hepatic spectral Doppler signal the microbubble contrast agent Levovist was used. The presence or absence of flow following microbubble contrast was evaluated against arteriography or repeated Doppler US findings. A total of 794 surveillance Doppler US examinations were performed in 231 patients. Hepatic artery flow was demonstrated in 759 of 794 (95.6%) examinations. Microbubble ultrasound contrast was administered in 31 patients (35 studies) with suspected hepatic artery thrombosis. Following microbubble US contrast the hepatic artery could not be demonstrated in 13 of 35 (37.1%) studies (12 patients). Eight patients had arteriography: there was hepatic artery thrombosis in 7 patients and 1 patient had a patent, highly attenuated artery. Detection of a patent hepatic artery increased from 759 of 794 (95.6%) to 781 of 794 (98.4%) with the addition of microbubble contrast. Upon independent reading of the data, the degree of operator confidence in the assessment of the hepatic artery patency prior to microbubble contrast was 4.7 (CI 1.92-7.5) but rose to 8.45 (CI 7.06-9.84) following microbubble contrast ( p<0.0001). In 22 of 35 (62.9%) of studies arteriography could potentially have been avoided. Ultrasound microbubble contrast media may reduce the need for invasive arteriography in the assessment of suspected hepatic artery thrombosis. PMID- 14530999 TI - Cerebral venous sinus thrombosis. AB - A comprehensive synopsis on cerebral venous thrombosis is presented. It emphasizes the various aetiologies, the wide clinical spectrum and the unpredictable outcome. Imaging techniques and pitfalls are reported and the therapeutic options are discussed. PMID- 14531000 TI - Can pre-operative contrast-enhanced dynamic MR imaging for prostate cancer predict microvessel density in prostatectomy specimens? AB - The aim of this study was to correlate quantitative dynamic contrast-enhanced MRI (DCE MRI) parameters with microvessel density (MVD) in prostate carcinoma. Twenty eight patients with biopsy-proven prostate carcinoma were examined by endorectal MRI including multiplanar T2- and T1-weighted spin-echo and dynamic T1-weighted turbo-FLASH MRI during and after intravenous Gd-DTPA administration. Microvessels were stained on surgical specimens using a CD31 monoclonal antibody. The MVD was quantified in hot spots by counting (MVC) and determining the area fraction by morphometry (MVAF). The DCE MRI data were analyzed using an open pharmacokinetic two-compartment model. In corresponding anatomic locations the time shift (Deltat) between the beginning of signal enhancement of cancer and adjacent normal prostatic tissue, the degree of contrast enhancement and the contrast exchange rate constant (k21) were calculated. The MVC and MVAF were elevated in carcinoma (p<0.001 and p=0.002, respectively) and correlated to k21 (r=0.62, p<0.001 and r=0.80, p<0.001, respectively). k21-values of carcinoma were significantly higher compared with normal peripheral but not central zone tissue. Deltat was longer in high compared with low-grade tumors (p=0.025). The DCE MRI can provide important information about individual MVD in prostate cancer, which may be helpful for guiding biopsy and assessing individual prognosis. PMID- 14531001 TI - Osteomyelitis in infants and children. AB - Infection of the pediatric skeletal system is a dramatic diagnostic and therapeutic challenge. The consequences of a missed diagnosis could lead to long lasting functional sequelae. This disease has multiple aspects according to the clinical presentation and evolution, the causative microorganism, the site of infection, and, finally, the patient's age and immunological status. The choice of the most appropriate imaging modality according to the clinical presentation is very important to assess the diagnosis, the extent of the disease, and to obtain the causative germ when needed; thus, plain films, ultrasound, nuclear medicine, computed tomography, and MR imaging are discussed, each of them having pros and cons. This multimodality approach of the musculoskeletal infections is mandatory to guide surgery, to deliver an appropriate antibiotic therapy, and to reduce the rate of long-term functional sequelae. PMID- 14531002 TI - Treatment of intermetatarsal Morton's neuroma with alcohol injection under US guide: 10-month follow-up. AB - Morton's neuroma (MN) is a frequent cause of metatarsalgia. The aim of our study was to evaluate the efficacy of neuroma alcohol-sclerosing therapy (NAST) under US guide in MN after a 10-month follow-up. Forty intermetatarsal neuromas underwent alcohol-sclerosing therapy after sonographic evaluation of their dimensions and echotexture. After subcutaneous anesthesia, a sclerosing solution composed of anesthetic (carbocaine-adrenaline 70%) and ethylic alcohol (30%) was injected inside the mass under US guidance. The procedure was repeated at intervals of 15 days until the resolution of the symptoms. A total or partial symptomatic relief was obtained in 36 cases (90%). No procedure-related complications were observed. Transitory plantar pain, due to the flogistic reaction induced by the sclerosing solution, occurred in 6 cases (15%). The 10 month follow-up revealed a 20-30% mass volume reduction and an adiposus-like change in echotexture. In the 4 cases (10%) of therapeutic failure, the preliminary sonography demonstrated a hypoechoic echotexture with a strong US beam attenuation corresponding to a highly fibrous neuroma after surgical resection. The NAST is a feasible and cost-efficient procedure with high rates of therapeutic success. PMID- 14531003 TI - Upload capacity and time-to-display of an image Web system during simultaneous up and download processes. AB - The aim of this study was to assess the up- and download performance of an image Web system (IWS) during simultaneous up- and download procedures. Following preparatory tests, the upload capacity (UC) with and without simultaneous download and the time-to-display (TTD) with and without simultaneous upload were determined for different image, server and compression types, and with up to 16 concurrent clients. The UC varied between 1.7-5.5 Gigabyte per hour (GB/h) and was slightly influenced by a simultaneous download; however, the TTDs were substantially prolonged during an upload and only below 5 s with up to four to six concurrent clients. The choice of image and server type had a strong impact on UC and TTD. Lossy primary compression proved slightly superior. An effective UC of approximately 4.0 GB/h or 96 Gigabyte per day (GB/day) can be achieved, which appears suitable even for large institutions. Because of its substantial influence on UC and TTD during simultaneous up- and download, the server hardware should be equipped with two processors and 1 GB RAM. Lossy primary compression may be used with slight performance benefits when full-resolution images are not required. The upload is a time-demanding process, and it is possible that during peak hours the waiting times are unacceptable for clinicians. PMID- 14531004 TI - MR appearance of cartilage defects of the knee: preliminary results of a spiral CT arthrography-guided analysis. AB - The aim of this study was to determine signal intensity patterns of cartilage defects at MR imaging. The MR imaging (3-mm-thick fat-suppressed intermediate weighted fast spin-echo images) was obtained in 31 knees (21 male and 10 female patients; mean age 45.5 years) blindly selected from a series of 252 consecutive knees investigated by dual-detector spiral CT arthrography. Two radiologists determined in consensus the MR signal intensity of the cartilage areas where cartilage defects had been demonstrated on the corresponding reformatted CT arthrographic images. There were 83 cartilage defects at spiral CT arthrography. In 52 (63%) lesion areas, the MR signal intensity was higher than that of adjacent normal cartilage with signal intensity equivalent to (n=31) or lower than (n=21) that of articular fluid. The MR signal intensity was equivalent to that of adjacent normal cartilage in 17 (20%) lesion areas and lower than that of adjacent cartilage in 8 (10%) lesion areas. In 6 (7%) lesion areas, mixed low and high signal intensity was observed. The MR signal intensity of cartilage defects demonstrated on spiral CT arthrographic images varies from low to high on fat suppressed intermediate-weighted fast spin-echo MR images obtained with our equipment and MR parameters. PMID- 14531005 TI - Preoperative detection of hepatocellular carcinoma: comparison of combined contrast-enhanced MR imaging and combined CT during arterial portography and CT hepatic arteriography. AB - The aim of this study was to compare Gd-DTPA-enhanced dynamic MR images, superparamagnetic iron oxide (SPIO)-enhanced MR images, combined Gd-DTPA-enhanced dynamic and SPIO-enhanced MR images, vs combined CT arterial portography (CTAP) and CT hepatic arteriography (CTHA), in the detection of hepatocellular carcinoma (HCC) using receiver operating characteristic (ROC) analysis. Twenty-four patients with 38 nodular HCCs (5-60 mm, mean 23.0 mm) were retrospectively analyzed. Image reviews were conducted on a liver segment-by-segment basis. A total of 192 segments, including 36 segments with 38 HCC, were reviewed independently by three radiologists. Each radiologist read four sets of images (set 1, unenhanced and Gd-DTPA-enhanced dynamic MR images; set 2, unenhanced and SPIO-enhanced MR images; set 3, combined Gd-DTPA-enhanced dynamic and SPIO enhanced MR images; set 4, combined CTAP and CTHA). To minimize any possible learning bias, the reviewing order was randomized and the reviewing procedure was performed in four sessions at 2-week intervals. The diagnostic accuracy (A(z) values) for HCCs of combined CTAP and CTHA, combined Gd-DTPA-enhanced dynamic and SPIO-enhanced MR images, Gd-DTPA-enhanced dynamic MR images, and SPIO-enhanced MR images for all observers were 0.934, 0.963, 0.878, and 0.869, respectively. The diagnostic accuracy of combined CTAP and CTHA and combined Gd-DTPA-enhanced dynamic and SPIO-enhanced MR images was significantly higher than Gd-DTPA enhanced dynamic MR images or SPIO-enhanced MR images ( p<0.005). The mean specificity of combined CTAP and CTHA (93%) and combined Gd-DTPA-enhanced dynamic and SPIO-enhanced MR images (95%) was significantly higher than Gd-DTPA-enhanced dynamic MR images (87%) or SPIO-enhanced MR images (88%; p<0.05). Combined Gd DTPA-enhanced dynamic and SPIO-enhanced MR images may obviate the need for more invasive combined CTAP and CTHA for the preoperative evaluation of patients with HCC. PMID- 14531007 TI - Potential applications of three-dimensional ultrasound in the pediatric urinary tract: pictorial demonstration based on preliminary results. AB - The aim of this study was to describe the potential of three-dimensional ultrasound (3DUS) in the urinary tract of neonates, infants, and children. The potential applications are illustrated based on our experience in 80 patients using two different 3DUS techniques. Various disease entities throughout the neonatal and pediatric age have been evaluated. The potential of 3DUS is discussed based on comparison with conventional 2DUS or other imaging (as clinically indicated), focused on the potentially improved renal parenchymal volume assessment. In our experience, 3DUS is feasible in neonates, infants, and children without sedation. It reduces imaging time, improves demonstration of complex anatomy and allows for evaluation of anatomy/pathology in any plane. The 3DUS improves volume assessment and follow-up comparison by offering an improved standardization and documentation. Rendered views of the dilated collecting system enable a comprehensive demonstration of hydronephrosis similar to intravenous urography or MR urography images. Additionally, 3DUS offers an ideal tool for training and education. Yet, limitations have to be acknowledged: areas inaccessible for 2DUS; poor quality of the original 2DUS acquisition; limited resolution; patient motion and breathing; cardiac pulsation creating artifacts and misregistration; equipment cost; lack of 3D DICOM standards creating problems with data storage; as well as system-inherent technical limitations. Nevertheless, the 3DUS holds the potential to become a valuable additional imaging tool for sonographic evaluation of the pediatric urinary tract. PMID- 14531008 TI - MR imaging of the contralateral breast in patients after breast-conserving therapy. AB - A study was undertaken to assess the diagnostic accuracy of contrast-enhanced MR mammography (MRM) of the contralateral breast in patients treated by breast conserving therapy previously. A total of 119 patients underwent 145 standardized dynamic MR studies (1 T, T1-weighted 3D FLASH, 0.2 mmol Gd-DTPA/kg body weight). We retrospectively evaluated the results of conventional methods and MRM. A total of 11 contralateral carcinomas were present (detection rate 9%). The interval between treatment of the first primary and identification of contralateral malignancy was 9-80 months (mean 33 months). The MRM allowed detection of four otherwise occult malignancies. One of 11 cancer was missed on MRM due to benign appearance of enhancement. Compared with conventional methods MRM improved sensitivity (91 vs 64%) and specificity (90 vs 84%), respectively. This study suggests that additional MRM of the contralateral breast increases the diagnostic accuracy not only by enhancing the detection of second cancers but also by reducing false-positive results. PMID- 14531009 TI - Spine fractures in falling accidents: analysis of multidetector CT findings. AB - The purpose of the present study was to assess incidence, fracture type, and location of spine fractures due to falls. All emergency room CT requests during a time period of 26 months were reviewed retrospectively. Patients who had fallen or jumped and were initially examined with multidetector CT (MDCT) were included. The MDCT studies were evaluated by two radiologists for trauma location, fracture type, and multiple level involvement. A total of 237 patients (184 males, 53 female, age range 16-86 years, mean age 42 years) met the inclusion criteria. A total of 203 vertebral fractures were seen in 127 patients. Burst fracture (n=78) was the most frequent type of trauma, usually located in the thoracolumbar junction (50%). Also, compression fracture (n=52) was most common in the thoracolumbar junction (39%). Posterior column fracture (n=52) was most frequently detected in the cervical spine (40%). Multiple-level spine fractures were seen in 41 (32%) of the injured patients, of which 12 (29%) had fractures at noncontinuous levels. With increasing height the overall incidence of fractures increased, and burst fractures and multiple level spine fractures became more frequent. Age had no effect on fracture type or location. Spine fractures due to falls are common. Burst fracture is the most common fracture type and most frequently seen in the thoracolumbar junction. Multiple-level fractures were seen in 32% of the cases, of which 29% were seen at noncontinuous levels. Serious spine fractures are seen in all falling height and age groups. PMID- 14531012 TI - The fibrocartilaginous sesamoid: a cause of size and signal variation in the normal distal posterior tibial tendon. AB - The aim of this study was to investigate the presence of fibrocartilage within the distal posterior tibial tendon (PTT) before its division correlating with size and signal variation on MR images through a radio-anatomic and pathologic study. Eight fresh cadaveric feet underwent MR imaging were cut into 4-mm slices in the axial plane. The PTT specimens were harvested at the tendon distal portion before its division and sent to pathology. Thirty-three asymptomatic subjects underwent axial double-echo turbo-spin-echo MR imaging. Proximal and distal PTT signal and diameter were evaluated. In cadavers, every PTT flared distally. Intratendinous fibrocartilage and ossified sesamoid were found in, respectively, 87.5 and 12.5% of the cases. Distal PTT flaring was demonstrated in 100% of the asymptomatic subjects (mean diameter 8 mm). An intratendinous high signal intensity on proton-density-weighted images and sesamoid bone were evidenced in, respectively, 36 and 33% of the cases. Proximally, PTT presented a 4-mm mean diameter and was hypointense in 100% of the cases. Only one accessory navicular bone was detected. Laterally off-centered increased intratendinous signal intensity as well as PTT distal widening with otherwise normal MR imaging features are related to an intratendinous fibrocartilage. PMID- 14531010 TI - Testicular microlithiasis: clinical significance and review of the literature. AB - Testicular microlithiasis (TM) is an uncommon condition characterized by calcium deposits within the seminiferous tubules. On ultrasound (US), it is seen as multiple, uniform, nonshadowing echogenic foci in the testis. Although its true prevalence in the general population is still unknown, reported prevalences range from 0.6 to 9%. The TM is often associated with germ cell tumor (GCT) or intratubular germ cell neoplasia. The incidence of GCT in patients with TM was reported as 6-46%. There are several reports demonstrating interval development of GCT in patients with TM. These may suggest a premalignant nature of TM; however, more recent studies show a lower incidence of associated GCT and no interval development of tumor in relatively longer duration follow-up. Additionally, previously reported cases of interval tumor development had predisposing factors for testicular GCT. According to the recent literature, it is suggested that both TM and testicular GCT may be caused by a common defect, such as tubular degeneration, and TM may present as a marker for such abnormalities; however, because of a high incidence of association with GCT, it is prudent to follow up patients with TM with physical examination and US at least annually and to encourage self-examination. The routine use of biochemical tumor markers, abdominal and pelvic CT, or testicular biopsy does not seem to be justified. PMID- 14531013 TI - Evaluation of MR cisternography of the cerebellopontine angle using a balanced fast-field-echo sequence: preliminary findings. AB - We evaluated the feasibility of MR cisternography by the balanced fast-field-echo (bFFE) sequence, comparing with that by a turbo-spin-echo (TSE) sequence, for cerebellopontine angle lesions on a 1.5-T imager (Gyroscan Intera, Philips, Best, The Netherlands). The bFFE MR cisternograms depicted target cranial nerves with less cerebrospinal fluid pulsation artifacts than TSE cisternograms and visualized an acoustic schwannoma in 6 of 44 patients with suspicion and a causative vessel of hemifacial spasm in all of 3 patients in a short scanning time (1 min 53 s). The bFFE sequence can be promising for MR cisternography in the diagnosis of cerebellopontine angle lesions. PMID- 14531014 TI - Image fusion of thallium-201 SPECT and MR imaging for the assessment of recurrent head and neck tumors following flap reconstructive surgery. AB - The aim of this study was to assess the value of fused MR and Tl-201 single photon emission computed tomography (SPECT) images in the diagnosis of recurrent head and neck tumors in patients after flap reconstruction surgery. Twenty-four patients after resection of primary head and neck tumors with flap reconstruction were suspected of having recurrent tumor by follow-up MR examination. Both MR examination and Tl-201 SPECT were prospectively performed to produce fused images. For qualitative analysis, two independent readers separately evaluated the existence of tumor recurrence in the fused images. The Tl-201 uptake of the lesion (Tl index) was also quantitatively compared with that of the normal nuchal muscles. Eighteen patients were histologically proved as having recurrence. The remaining 6 patients, false positive on MRI alone, had non-recurrence. Using the fused images, false positive was found in 1 case for one reader and 2 cases for the other reader. The Tl index of recurrent tumors was significantly higher (p < 0.001) than that of non-recurrent mass lesions. In the assessment of recurrent tumors following flap reconstruction surgery in the head and neck, the use of fused MRI and Tl-201 SPECT images can reduce the number of false positives. PMID- 14531015 TI - Involvement of nuclear factor-kappaB in the inhibition of interleukin-12 production from mouse macrophages by baicalein, a flavonoid in Scutellaria baicalensis. AB - Pharmacological inhibition of interleukin-12 (IL-12) production may be a therapeutic strategy for preventing development and progression of disease in experimental models of autoimmunity. In this study we investigated the effects of baicalein, a flavonoid present in the root of Scutellaria baicalensis, on the production of IL-12 from mouse macrophages stimulated with lipopolysaccharide (LPS). Baicalein potently inhibited the LPS-induced IL-12 production from both primary macrophages and RAW264.7 monocytic cell-line in a dose-dependent manner (the IC50 values were 43.7 and 17.4 microM, respectively). The effect of baicalein on IL-12 gene promoter activation was analyzed by transfecting RAW264.7 cells with IL-12 gene promoter/luciferase constructs. The repressive effect mapped to a region in the IL-12 gene promoter containing a binding site for NF kappaB. Furthermore, activation of macrophages by LPS resulted in markedly enhanced binding activity to the NF-kappaB site, which significantly decreased upon addition of baicalein, indicating that baicalein inhibited IL-12 production in LPS-activated macrophages via inhibition of NF-kappaB binding activity. PMID- 14531016 TI - Inhibitory effect of jujuboside A on glutamate-mediated excitatory signal pathway in hippocampus. AB - Jujuboside A (JuA) is a main component of jujubogenin extracted from the seed of Ziziphus jujuba Mill var spinosa (Bunge) Hu ex H F Chou (Ziziphus), which is widely used in Chinese traditional medicine for the treatment of insomnia and anxiety. Previously, we reported the inhibitory effects of JuA on hippocampal formation in vivo and in vitro, the present study was carried out to examine the effects of JuA on glutamate (Glu)-mediated excitatory signal pathway in hippocampus. Microdialysis coupled with high-performance liquid chromatography (HPLC) was used to monitor the changes of Glu levels in the hippocampus induced by penicillin sodium, or a mixture of penicillin sodium and JuA. The results showed that penicillin increased the hippocampal Glu concentration (p < 0.01) and a high dose of JuA (0.1 g/L) significantly blocked penicillin-induced Glu release (p < 0.05). Moreover, the effect of JuA on intracellular Ca2+ changes after the stimulation by Glu was studied in cultured hippocampal neurons with confocal laser scanning microscope (CLSM). It was found that Glu (0.5 mM) induced an intracellular [Ca2+]i increase (p < 0.01), and JuA significantly inhibited the Glu-induced Ca2+ increase. The calmodulin (CaM) antagonist trifluoperazine (TFP) showed a similar inhibitory effect as JuA. These observations suggested that JuA has inhibitory effects on Glu-mediated excitatory signal pathway in hippocampus and probably acts through its anti-calmodulin action. PMID- 14531017 TI - Suppression of inducible nitric oxide production by indole and isothiocyanate derivatives from Brassica plants in stimulated macrophages. AB - In this study, the effects of bioactive compounds derived from vegetables of the Brassica genus (Brassicaceae) including 2-phenylethyl isothiocyanate (PEITC), indole-3-carbinol (I3C), and indolo[3,2- b]carbazole (ICZ), on the inhibition of NO production in RAW 264.7 cells were explored. The results indicated that PEITC and I3C inhibited lipopolysaccharide (LPS)- and interferon-gamma (IFN-gamma) induced NO production in RAW 264.7 cells, and this inhibition was in accordance with lowering the expression of iNOS protein and mRNA. On the contrary, ICZ, a derivative of I3C, had no significant effect on the stimulated NO production. In conclusion, the Brassica plants derivatives, PEITC and, to a lesser extent, I3C inhibit the LPS/IFN-gamma-induced NO production by lowering iNOS protein and mRNA expression in RAW 264.7 cells, in which the PEITC had a more potent inhibitory effect. Nevertheless, ICZ exhibits no inhibitory effect on the activated NO production (Indole-3-carbinol = indole-3-methanol). PMID- 14531018 TI - Anti-tumour activity of Digitalis purpurea L. subsp. heywoodii. AB - Recent research has shown the anticancer effects of digitalis compounds suggesting their possible use in medical oncology. Four extracts obtained from the leaves of Digitalis purpurea subsp. heywoodii have been assessed for cytotoxic activity against three human cancer cell lines, using the SRB assay. All of them showed high cytotoxicity, producing IC50 values in the 0.78 - 15 microg/mL range with the methanolic extract being the most active, in non toxic concentrations. Steroid glycosides (gitoxigenin derivatives) were detected in this methanolic extract. Gitoxigenin and gitoxin were evaluated in the SRB assay using the three human cancer cell lines, showing IC50 values in the 0.13 - 2.8 microM range, with the renal adenocarcinoma cancer cell line (TK-10) being the most sensitive one. Morphological apoptosis evaluation of the methanolic extract and both compounds on the TK-10 cell line showed that their cytotoxicity was mediated by an apoptotic effect. Finally, possible mechanisms involved in apoptosis induction by digitalis compounds are discussed. PMID- 14531019 TI - Cytotoxicity and anti-hepatitis B virus activities of saikosaponins from Bupleurum species. AB - Saikosaponins, the main active constituents of Bupleurum spp., have been shown to possess immunomodulatory, hepatoprotective, anti-tumor and anti-viral activities. In this study, saikosaponins a, c and d were evaluated for cytotoxicity and anti hepatitis B virus ( HBV) activities. Results showed that, with the exception of saikosaponins a and d, HBV-transfected human hepatoma cells (2.2.15 cells) cultured with saikosaponin c showed a significantly lower level of HBeAg in culture medium. Saikosaponin c also possessed activity in inhibiting HBV DNA replication; this inhibitory effect was not due to the cytotoxicity of saikosaponin c or its effect on 2.2.15 cell proliferation. Although saikosaponin d exhibited cytotoxicity on 2.2.15 cells, it failed to inhibit HBV multiplication. The cytotoxicity of saikosaponin d against HepG2 human hepatocellular carcinoma cells was due to the induction of apoptosis through the activation of caspases 3 and 7, which subsequently resulted in poly-ADP-ribose polymerase (PARP) cleavage. DNA fragmentation was clearly noted at more than 6 h after HepG2 cells exposure to saikosaponin d. The present study concludes that saikosaponin c exhibits anti-HBV activity and saikosaponin d possesses potent cytotoxicity against human hepatocellular carcinoma cells. PMID- 14531020 TI - Anti-inflammatory effect of the oligomeric stilbene alpha-Viniferin and its mode of the action through inhibition of cyclooxygenase-2 and inducible nitric oxide synthase. AB - The anti-inflammatory activity of alpha-viniferin, a trimer of resveratrol, has been demonstrated in an animal model of carrageenin-induced paw edema, and inhibitory effects of the compound on cyclooxygenase (COX) and inducible nitric oxide synthase (iNOS) have been investigated in order to understand the mode of the observed action. alpha-Viniferin at doses > 30 mg/kg ( p. o.) or > 3 mg/kg ( i. v.) showed significant anti-inflammatory activity on carrageenin-induced paw edema in mice. alpha-Viniferin showed an inhibitory effect with an IC (50) value of 4.9 microM on COX-2 activity but a very weak inhibitory effect with 55.2 +/- 2.1 % of the control (100 %) at 100 microM on COX-1 activity. alpha-Viniferin at doses of 3 microM to 10 microM inhibited the synthesis of COX-2 transcript in lipopolysaccharide (LPS)-activated murine macrophages Raw264.7. alpha-Viniferin showed an IC50 value of 2.7 microM on nitric oxide (NO) production in LPS activated Raw264.7 cells when alpha-viniferin and LPS were treated simultaneously, but did not inhibit the NO production when alpha-viniferin was treated at 12 h after LPS stimulation. alpha-Viniferin inhibited synthesis of iNOS transcript with an IC50 value of 4.7 microM. Consequently, the inhibitory effect of alpha-viniferin on the release of prostanoids and NO could play an important role to show anti-inflammatory action. PMID- 14531021 TI - Albaconol from the mushroom Albatrellus confluens induces contraction and desensitization in guinea pig trachea. AB - The contraction and desensitization induced by albaconol and the influence of capsazepine, capsaicin and extracellular Ca2+ were investigated to see whether the actions were mediated via a specific VR receptor in guinea pig trachea spiral strips in vitro. Both albaconol and capsaicin were contractors of tracheal smooth muscle, but albaconol was not so potent as capsaicin, with -log (M) EC50 values of 4.23 +/- 0.18 (n = 10) and 7.33 +/- 0.21 (n = 10) respectively. 2.5 microM capsazepine competitively antagonized the contractile response to albaconol and capsaicin. Albaconol increased the contraction induced by a low dose of capsaicin (10(-10) to 10(-9) M), but non-competitively antagonized the contraction induced by a high dose of capsaicin (10(-8) to 10(-3) M). Either albaconol (1 or 100 mM) or capsaicin (3 or 10 microM) was able to desensitize the isolated guinea pig bronchi to subsequent addition of albaconol. Capsazepine (5.0 microM) significantly prevented the desensitization induced by either albaconol (1 or 100mM) or capsaicin (3 or 10 microM). Extracellular Ca2+ was essential for albaconol to induce excitation, but it did not affect albaconol- or capsaicin induced desensitization. In summary, the results from the present study suggest that albaconol induces contraction and desensitization of guinea pig trachea in vitro as a partial agonist for VR. PMID- 14531023 TI - Gene expression changes in the human fibroblast induced by Centella asiatica triterpenoids. AB - The molecular pathways underlying the diverse biological activity of the triterpeniod compounds isolated from the tropical medicinal plant Centella asiatica were studied with gene microarrays and real-time reverse transcription polymerase chain reaction (real-time RT-PCR) to quantify the expression of 1053 human genes in human fibroblasts. Fibroblast cells grown in culture were used as a model system to evaluate the stimulation of wound healing by titrated extract from Centella asiatica (TECA) as well as by the four principal triterpenoid components of Centella. TECA treatment effects the expression of genes involved in angiogenesis and the remodeling of extracellular matrix, as well as diverse growth factor genes. The extent of expression change of TNFAIP6, an extracellular hyaluronan binding protein, was found to be largely dose-dependent, to respond most strongly to the free acids asiatic acid and madecassic acid, and to increase in expression over 48 hours of treatment. These results show that Centella triterpenes evoke a gene-expression response consistent with their prevailing medical uses in the treatment of connective tissue disorders such as wound healing and microangiopathy. The identification of genes modulated by these compounds provides the basis for a molecular understanding of Centella's bioactivity, and opportunities for the quantitative correlation of this activity with clinical effectiveness at a molecular level. PMID- 14531022 TI - In vitro antiplasmodial activity of abietane and totarane diterpenes isolated from Harpagophytum procumbens (devil's claw). AB - The development of drug resistance and resurgence of malaria has highlighted the need for new chemically diverse antimalarial drugs. This study investigates Harpagophytum procumbens DC. as a source of antiplasmodial hit compounds. The roots of wild harvested plants as well as the aerial sections, seeds and roots of cultivated H. procumbens were evaluated for in vitro antiplasmodial activity. Bioassay-guided fractionation of the petroleum ether root extract yielded two diterpenes, (+)-8,11,13-totaratriene-12,13-diol (1) and (+)-8,11,13-abietatrien 12-ol (2). Compounds 1 and 2 displayed significant (IC50 < 1 microg/mL) in vitro antiplasmodial activity against a chloroquine-resistant (K1) and -sensitive (D10) strain of Plasmodium falciparum, and low cytotoxicity (SI > 65) against two mammalian cell lines (CHO and HepG2). It was found that 1 and 2 did not modify the erythrocyte shape, which in conjunction with the cytotoxicity results, indicates selective antiplasmodial activity. PMID- 14531024 TI - A phytochemical study of lignans in whole plants and cell suspension cultures of Anthriscus sylvestris. AB - In the roots of Anthriscus sylvestris 12 different lignans were detected. Arctigenin, dimethylmatairesinol, dimethylthujaplicatin, podophyllotoxin, 7 hydroxyyatein and 7-hydroxyanhydropodorhizol have not been previously reported to be present in A. sylvestris. In the cell suspension cultures, which were initiated for this study, trace amounts of deoxypodophyllotoxin could be detected. With these cell suspension cultures we carried out feeding experiments using deoxypodophyllotoxin, yatein and, anhydropodorhizol. Yatein had a toxic effect on the cell cultures and was, like anhydropodorhizol, not converted into any detectable product. Deoxypodophyllotoxin, in contrast, was converted into podophyllotoxin, yielding significantly higher concentration than measured in whole plants. PMID- 14531025 TI - The bioconversion process of deoxypodophyllotoxin with Linum flavum cell cultures. AB - The in vitro cell suspension culture of Linum flavum is able to convert high amounts of the 2,7'-cyclolignan deoxypodophyllotoxin into 6 methoxypodophyllotoxin 7- O-glucoside. We studied this conversion in detail by monitoring the intermediates and side-products after feeding different concentrations of deoxypodophyllotoxin. At a low concentration (0.1 mM) deoxypodophyllotoxin is rapidly converted into 6-methoxypodophyllotoxin 7- O glucoside, 6-methoxypodophyllotoxin and traces of beta-peltatin and podophyllotoxin. The feeding of 0.5 and 2.0 mM also shows a rapid conversion into 6-methoxypodophyllotoxin 7- O-glucoside, but a delayed formation of 6 methoxypodophyllotoxin and beta-peltatin. By using different extraction methods we delivered proof in favour of the hypothesis that a part of the deoxypodophyllotoxin after uptake is temporarily stored as beta-peltatin glucoside. PMID- 14531026 TI - New eremophilane sesquiterpenes from Cacalia ainsliaeflora. AB - One known eremophilanolide, 3beta-angeloyloxy-8beta,10beta-dihydroxy-6beta methoxyeremophilenolide (1) and seven new eremophiane sesquiterpenes, 3beta angeloyloxy-6beta-ethoxy-8beta,10beta-dihydroxyeremophilenolide (2), 3beta,6beta diangeloyloxy-10alpha-hydroxy-8alpha-methoxyeremophilenolide, 6beta-angeloyloxy 8beta,10beta-dihydroxy-3-oxo-eremophilenolide, 6beta,8beta,10beta-trihydroxy-3 oxo-eremophilenolide, 3beta-angeloyloxy-6beta,10beta-dihydroxy-eremophila 7(11),8(9)-dien-8,12-olide (6), 3beta-angeloyloxy-10beta-hydroxy-6beta-methoxy eremophila-7(11),8(9)-dien-8,12-olide, 3beta-angeloyloxy-8-oxo-eremophila-6.9 dien-12-oic acid ethyl ester, were isolated from the roots of Cacalia ainsliaeflora. Their structures were elucidated by spectroscopic methods. Compounds 1, 2 and 6 were assayed against both Gram-positive and Gram-negative bacteria. No positive activities were observed. PMID- 14531027 TI - Cytotoxic saponins from Schefflera fagueti. AB - Six new lupane (1 - 4) and oleanane saponins (5 and 6) were isolated from the aerial parts of Schefflera fagueti Baill. (Araliaceae). Their structures were determined by 2D-NMR spectroscopy (DQF-COSY, 1D-TOCSY, 2D-HOHAHA, 1D-ROESY, HSQC, HMBC). The antiproliferative activity of compounds 1 - 6 and of their prosapogenins (1a - 6a) was evaluated using three continuous murine and human culture cell lines J774, HEK-293, WEHI-164. Oleanane saponins 5 and 6 were the most active, showing significant inhibitory effects on all cell lines, while their prosapogenins 5a and 6a demonstrated minor activity. PMID- 14531028 TI - Anti-platelet aggregation and chemical constituents from the rhizome of Gynura japonica. AB - A novel quinonoid terpenoid, (-)- alpha-tocospirone, a new chromanone, (-) gynuraone, as well as three new steroids, (22 E,24 S)-7 alpha hydroperoxystigmasta-5,22-dien-3 beta-ol, (22 E,24 S)-stigmasta-1,4,22-trien-3 one, and (24 R)-stigmasta-1,4-dien-3-one, together with 15 known components, were isolated from the rhizome of Formosan Gynura japonica. Structures of the new compounds were determined through spectral analyses and chemical evidence. Several of the isolated compounds, caryophyllene oxide, 6-acetyl-2,2 dimethylchroman-4-one, vanillin, 2,6-dimethoxy-1,4-benzoquinone, and benzoic acid exhibited significant anti-platelet aggregation activity in vitro. PMID- 14531030 TI - Identification of an anti-inflammatory principle from the stem bark of Millettia versicolor. AB - The dichloromethane-soluble fraction of the methanol extract of the stem bark of Millettia versicolor Welw. (Leguminosae) has been shown to possess anti inflammatory activity. The chromatographic fractionation and subsequent analysis of the spectroscopic data of this extract led to the isolation and identification of 2-acetyl-7-methoxynaphtho[2,3- b]furan-4,9-quinone (1) along with two known quinones. Pharmacological data demonstrate that compound 1 has relevant anti inflammatory properties whereas the other two isolated compounds do not. PMID- 14531029 TI - Dual effect of saikogenin D: in vitro inhibition of prostaglandin E2 production and elevation of intracellular free Ca2+ concentration in C6 rat glioma cells. AB - To clarify the pharmacological profile of saikogenin D, we examined the effect of saikogenin D on prostaglandin E2 (PGE2) production and intracellular free Ca2+ concentration ([Ca2+]i) in C6 rat glioma cells. Saikogenin D (1-20 microM) inhibited PGE2 production induced by the Ca2+ ionophore A23187 in a concentration dependent manner with the IC50 of about 3 microM. Saikogenin D did not affect the conversion of arachidonic acid into PGE2 in microsomal preparations. On the other hand, saikogenin D elevated [Ca2+]i in a concentration-dependent manner (10-100 microM) with the EC50 value of about 35 microM in the presence or absence of extracellular Ca2+. These results suggest that saikogenin D possesses a dual effect: an inhibition of A23187-induced PGE2 production without a direct inhibition of cyclooxygenase activity; and an elevation of [Ca2+]i that is attributed to Ca2+ release from intracellular stores. PMID- 14531031 TI - The xanthones gentiacaulein and gentiakochianin are responsible for the vasodilator action of the roots of Gentiana kochiana. AB - Gentiana kochiana Perr. et Song. (Gentianaceae), a plant used in the traditional medicine of Tuscany (Italy) as antihypertensive remedy, exerts a vasodilator action on in vitro aortic rings that is probably linked to the blocking of the ryanodine-sensitive Ca++ channels. In the present study, three known xanthones were isolated from the crude methanolic extract of the roots: gentiacaulein, gentiakochianin, and swertiaperennin. The first two showed a vasorelaxing activity in rat aortic preparations, pre-contracted by 3 microM norepinephrine (pIC50 = 5.00 +/- 0.032 for gentiacaulein, pIC50 = 4.95 +/- 0.068 for gentiakochianin), 20 mM KCl (pIC50 = 4.90 +/- 0.15 for gentiacaulein; 4.59 +/- 0.069 for gentiakochianin), or 5 mM caffeine; on the contrary, in the same conditions, swertiaperennin did not show any vasodilator effect. In conclusion, gentiacaulein and gentiakochianin seem to be the compounds responsible for the vasorelaxing properties of the crude extract of Gentiana kochiana roots. PMID- 14531032 TI - Composition and antibacterial activity of the essential oils from Zanthoxylum rhoifolium. AB - The essential oils from the aerial parts of leaves, fruits and flowers of Zanthoxylum rhoifolium of Southern Brazil (Rio Grande do Sul), were analysed by GC, GC/MS, and chiral phase gas chromatography (CPGC). Forty-eight compounds were identified from the essential oils. The major constituents of the essential oil of the leaves were germacrene D (34 %) and bicyclogermacrene (23 %) and of the fruits, menth-2-en-1-ol (46.2 %), beta-myrcene (30.2 %), (-)-linalool (15 %) and (-)-alpha-terpineol (8.45 %). beta-Myrcene (65 %) and menth-2-en-1-ol (5.4 %) dominate the essential oil of the flowers. The oils of the leaves and fruits were bioactive with antibacterial activity against Staphylococcus aureus (Gram positive), and Klebsiella pneumoniae and Salmonella setubal bacteria (Gram negative) microorganisms, while the essential oil of the flowers was inactive. PMID- 14531033 TI - Antitubercular cassane furanoditerpenoids from the roots of Caesalpinia pulcherrima. AB - Activity-guided fractionation of a root extract of Caesalpinia pulcherrima led to the isolation of two cassane-furanoditerpenoids, 6 beta-benzoyl-7 beta hydroxyvouacapen-5 alpha-ol (1) and 6 beta-cinnamoyl-7beta-hydroxyvouacapen-5 alpha-ol (2). Compound 2 showed strong antitubercular activity with a minimum inhibitory concentration (MIC) of 6.25 microg/mL, whereas the benzoyl analogue (1) was less active (MIC 25 microg/mL). Both compounds expressed moderate cytotoxic activity towards KB (human oral carcinonoid cancer), BC (human breast cancer) and NCl-H187 (small cell lung cancer) cell lines. PMID- 14531034 TI - In vitro micropropagation, differentiation of aerial bulbils and tubers and diosgenin content in Dioscorea bulbifera. AB - Dioscorea bulbifera could be micropropagated through nodal segments and bulbils. The best medium for regeneration and bulbil differentiation was MS + 0.5 microM IAA (indole-3-acetic acid) + 20.0 microM Kn (kinetin) + 500 mg/L CH (casein hydrolysate) + activated charcoal (20 %). Diosgenin content was maximum in regenerants grown on MS + 5.0 microM IAA + 20.0 microM Kn + 500 mg/L CH. T.s of bulbils could also be used for direct plantlet differentiation as well as bulbil differentiation on MS + 10.0 microM IAA + 20.0 microM Kn + (in mg/L) 30 each of Asp (asparagine) + Arg (arginine) + Gln (glutamine) + 10 Ad (adenine) + 500 CH + 10 Cyst hyd (cysteine hydrochloride). Diosgenin yield in plantlets reached a maximum after 20 weeks. The results indicate that micropropagation, bulbil formation and tuberisation can be achieved in vitro in D. bulbifera, hitherto a less exploited plant, and can further be used for obtaining enhanced levels of diosgenin. PMID- 14531035 TI - Halophilols A and B, two new stilbenes from Iris halophila. AB - A new monomeric stilbene, halophilol A (1), and a new tetrastilbene, halophilol B (2), along with three known oligostilbenes were isolated from the seeds of Iris halophila. Their structures were established on the basis of the spectral data. The oligostilbene skeleton is encountered for the first time in the Iridaceae family. Bioactivity tests showed that 1 had moderate cytotoxicity against KB and HMEC cell lines (IC50 = 17.28 microM, 22.47 microM respectively), while 2 was inactive. PMID- 14531036 TI - Germacrane sesquiterpene esters from Salvia roborowskii. AB - From the whole plant of Salvia roborowskii, four new germacrane sesquiterpene esters, 3beta,6beta,8alpha-triacetoxy-4beta,5alpha-epoxy-1-oxogermacr-10(14)-ene, 3beta,6beta,8alpha-triacetoxy-4beta,5alpha-epoxygermacr-1 (10) E-ene, 3beta,6beta,8alpha-triacetoxy-4beta,5alpha:1alpha,10beta-diepoxygermacrane and 6beta-acetoxyglechomafuran were isolated, and their structures were elucidated by spectroscopic methods including 2D NMR techniques (1H-1H COSY and 1H-1H NOESY for 1). PMID- 14531038 TI - Dopamine transporter imaging. PMID- 14531039 TI - Dopamine transporter: basic aspects and neuroimaging. AB - The plasma membrane dopamine transporter (DAT) is found exclusively in dopamine neurones and seems to be the defining molecule of the dopamine neurone. It provides effective control over the intensity of dopamine-mediated signalling by recapturing the neurotransmitter released by presynaptic neurones. Positron emission tomography (PET) and single-photon emission computed tomography (SPECT) represent unique techniques for assessing in vivo DAT distribution in humans and offer reliable methods for studying nigrostriatal dopaminergic function in health and disease. The characteristics of different DAT radiotracers, the modifying influences of factors such as age, gender, smoking habit, and dopaminergic drugs on DAT transporters as well as their implication in evaluation of neuroimaging studies are discussed. PMID- 14531040 TI - Role of dopamine transporter SPECT for the practitioner and the general neurologist. AB - The accurate clinical diagnosis of parkinsonism may be impeded by atypical presentations and confounding comorbidity. The presence of parkinsonism is misdiagnosed in up to a quarter of cases in general practice. Movement disorder specialists misdiagnose parkinsonian syndromes using histopathological findings as the "gold standard" in up to 10% of cases. Dopamine transporter SPECT represents a simple and fast method to confirm nigrostriatal degeneration in a given patient. This study provides several case reports to illustrate when dopamine transporter SPECT might be carried out and discusses whether dopamine transporter SPECT should be used in primary health care practice or by general neurologists in uncertain cases. Ideally, all possible cases of parkinsonism should be referred to a neurologist experienced in the field of movement disorders. If this could be achieved then the role of dopamine transporter SPECT in the general practitioner's or local neurologist's practice would be extremely limited. Future studies must clarify whether it is cost effective to generously perform dopamine transporter SPECTs to minimize the time until parkinsonism can be diagnosed. PMID- 14531041 TI - Role of dopamine transporter imaging in investigation of parkinsonian syndromes in routine clinical practice. AB - We discuss the potential role of dopamine transporter (DAT) imaging by single photon emission computed tomography (SPECT) as a biological marker for differentiating among parkinsonian disorders in routine clinical practice, and the implication for disease progression assessment is considered. Although clinical criteria enable accurate and reliable diagnosis of fully developed Parkinson's disease, there are several areas of diagnostic uncertainty relating to atypical parkinsonian disorders, isolated tremor symptoms not fulfilling essential tremor criteria, as well as drug-induced and psychogenetic parkinsonism. DAT-SPECT facilitates differential diagnosis of the different parkinsonian syndromes that cannot be reliably separated on clinical grounds and it can thus provide valuable diagnostic information in early disease. PMID- 14531042 TI - Role of dopamine transporter imaging in the diagnosis of atypical tremor disorders. AB - Dopamine transporter (DAT) imaging detects presynaptic dopamine neuronal dysfunction and thereby assists differentiation of conditions with and without dopamine deficit. In atypical tremor disorders, DAT imaging can differentiate between Parkinson's disease (PD), where dopamine deficit is demonstrated on DAT imaging, and essential tremor, where no dopamine deficit is found. DAT imaging may be particularly informative in monosymptomatic rest tremors, benign tremulous Parkinson's syndrome, and in the elderly in whom essential tremor may be accompanied by pseudoparkinsonism. PMID- 14531043 TI - DAT imaging in drug-induced and psychogenic parkinsonism. AB - Parkinson's syndrome (PS) is frequently encountered in disorders associated with prominent degeneration of the nigrostriatal pathway as in Parkinson's disease, multisystem atrophy, and progressive supranuclear palsy (presynaptic PS). Drug induced parkinsonism, a common, underdiagnosed health problem and psychogenic parkinsonism are causes of Parkinson's syndrome which, evidence suggests, occurs without degeneration of nigrostriatal structures. We review clinical features and results of DAT imaging in drug-induced parkinsonism and psychogenic parkinsonism. These two conditions normally give normal striatal DAT imaging results; an abnormal result in either case could exclude both conditions, corroborating a diagnosis of organic parkinsonism in uncertain cases. PMID- 14531044 TI - Dementia with Lewy bodies versus Alzheimer's disease: role of dopamine transporter imaging. AB - Dementia with Lewy bodies (DLB) and Alzheimer's disease (AD) are the common forms of dementia at post-mortem, but distinguishing between these two types of dementia is often very difficult during life. Ioflupane significantly improves the differentiation during life between DLB and AD patients. However, there is a trend for lower caudate uptake in DLB than PD and lower posterior/caudal putamen uptake in PD than in DLB. Further research is needed to test the hypothesis that dopaminergic degeneration may be different, at least regarding anatomical distribution, in DLB and PD. Furthermore, it is important to consider and discuss the potential for ioflupane in the diagnostic workup of patients with DLB. PMID- 14531045 TI - Significance of non-presynaptic SPECT tracer methods in Parkinson's disease. AB - Tropane-related tracers for SPECT and PET scanning have shown their value in assessing the in vivo striatal presynaptic status of the dopaminergic neurotransmitter system in man. Previously, there were a few other tracers suggested to be useful in the study of in vivo brain biochemistry in humans, particularly in patients afflicted with Parkinson's disease (PD) or parkinsonian syndromes. We summarize the work concerning the two main tracers applied in that context using single photon emission tomography (SPECT) scanning in patients with PD: IBZM and HMPAO. IBZM binds specifically to striatal dopamine D(2) receptors and HMPAO uptake is a measure of cerebral tissue perfusion. PMID- 14531046 TI - Correlation of dopamine transporter imaging with parkinsonian motor handicap: how close is it? AB - During the past decade, dopamine transporter (DAT) imaging with single photon emission computerized tomography (SPECT) or positron emission tomography (PET) has evolved as an objective in vivo marker of nigrostriatal neuron loss in Parkinson's disease (PD). We investigate the relationship between striatal DAT binding, measured with [(123)I]beta-CIT SPECT, and parkinsonian motor handicap in a sample of 59 PD patients with minimal to severe disability, and review published cross-sectional studies on the correlation between DAT imaging and motor symptoms in PD. Earlier studies as well as the present results show a good correlation between overall striatal DAT binding and global measures of disease severity such as the Hoehn and Yahr scale, the total score on the Unified Parkinson's Disease Rating Scale (UPDRS), and UPDRS activities of daily living, with a progressive decline of DAT binding with increasing disability. A number of studies found a significant inverse correlation of striatal DAT binding with UPDRS motor score. Bradykinesia, posture, gait, and other midline symptoms, such as speech and facial expression, compared with rigidity, seem to be more closely related to striatal DAT binding. By contrast, neither the severity of parkinsonian rest nor of action tremor is related to the degree of dopaminergic denervation as measured by DAT imaging. Motor symptoms in the clinically less affected body side show a closer correlation with striatal DAT binding than do symptoms occurring in the dominantly affected body side. The correlation of putamen and caudate DAT binding with parkinsonian motor handicap seems to be similar. Although there have been limited comparative studies applying [(18)F]fluorodopa PET and DAT imaging in the same group of PD patients, available data suggest that putamen [(18)F]fluorodopa uptake, when compared with striatal DAT binding, may be more closely related to parkinsonian motor handicap. PMID- 14531047 TI - Dopamine transporter imaging and SPECT in diagnostic work-up of Parkinson's disease: a decision-analytic approach. AB - As a diagnostic test for patients with suspected Parkinson's disease (PD), single photon emission computed tomography (SPECT) using [(123)I]FP-CIT tracer has better sensitivity but is more expensive than regular clinical examination (CE). Our objective was to evaluate the clinical and economic impacts of different diagnostic strategies involving [(123)I]FP-CIT SPECT. We developed a decision tree model to predict adequate treatment-month equivalents (ATME), costs, and incremental cost-effectiveness ratio (ICER) during a 12-month time horizon in patients with suspected PD referred to a specialized movement disorder outpatient clinic. In our cost- effectiveness analysis, we adopted the perspective of the German health care system and used data from a German prospective health care utilization study (n = 142) and published diagnostic studies. Compared strategies were CE only (EXAM+), SPECT only (SPECT+), SPECT following negative CE (SINGLE+), and SPECT following positive CE (DOUBLE+). Costs of SPECT amounted to euro;789 per investigation. Based on our model, expected costs (and ATME) were euro;946 (52.85 ATME) for EXAM+, euro;1352 (53.40 ATME) for DOUBLE+, euro;1731 (32.82 ATME) for SINGLE+, and euro;2003 (32.96 ATME) for SPECT+; performance of SPECT was induced in 0%, 54%, 56%, and 100% of the patients, respectively. DOUBLE+ was more effective and less expensive than SINGLE+ or SPECT+; thus these two do not offer reasonable choices. The ICER of DOUBLE+ compared to EXAM+ was euro;733 per ATME gained. In sensitivity analyses, the ICER of DOUBLE+ versus EXAM+ ranged from euro;63 to euro;2411 per ATME gained. Whether the diagnostic work-up of patients referred to a specialized movement disorder clinic with a high prevalence of PD should include [(123)I]FP-CIT SPECT depends on patient preferences and the decision maker's willingness to pay for adequate early treatment. SPECT should be used as a confirmatory test before treatment initiation and limited to patients with a positive test result in the clinical examination. These results should be adjusted to the specific setting and individual patient preferences. PMID- 14531048 TI - How valid is dopamine transporter imaging as a surrogate marker in research trials in Parkinson's disease? AB - To use functional imaging data as indices of disease progression, it is essential that methods are valid. It is not possible for that validation to come from pathology and the methods can only gain validity from the relationship to clinical indices of progression. Validity as a measurement of disease progression depends upon sensitivity to clinical progression, low scan-to-scan error, and resilience to confounding effects. Data from the available dopamine transporter (DAT) positron emission tomography (PET) and single photon emission computed tomography (SPECT) studies are examined for these three key factors. Presently, there are insufficient data to enable the satisfactory design of neuroprotection studies and to enable statements on their relevance to disease progression and neuroprotection. The recently completed CALM-PD neuroimaging study is examined. There is sufficient doubt surrounding the sensitivity, reproducibility, and confounding effects in this study that these data should not be used to formulate a decision on de novo therapy in Parkinson's disease. PMID- 14531049 TI - Dopamine transporter: basic science and human variation of a key molecule for dopaminergic function, locomotion, and parkinsonism. AB - We review the basic science of the dopamine transporter (DAT), a key neurotransmitter for locomotor control and reward systems, including those lost or deranged in Parkinson's disease (PD). Physiology, pharmaceutical features, expression, cDNA, protein structure/function relationships, and phosphorylation and regulation are discussed. The localization of DAT provides the best marker for the integrity of just the pre-synaptic dopaminergic systems that are most affected in PD. Its function is key for the actions of several toxins that provide some of the best current models for idiopathic parkinsonism, and its variation can clearly alter movement. The wealth of information about this interesting molecule that has been developed over the last 12 years has led to increased interest in DAT among workers interested in both normal and abnormal movement. PMID- 14531050 TI - Inclusion of the ligand field contribution in a polarizable molecular mechanics: SIBFA-LF. AB - To account for the distortion of the coordination sphere that takes place in complexes containing open-shell metal cations such as Cu(II), we implemented, in sum of interactions between fragments ab initio computed (SIBFA) molecular mechanics, an additional contribution to take into account the ligand field splitting of the metal d orbitals. This term, based on the angular overlap model, has been parameterized for Cu(II) coordinated to oxygen and nitrogen ligands. The comparison of the results obtained from density functional theory computations on the one hand and SIBFA or SIBFA-LF on the other shows that SIBFA-LF gives geometric arrangements similar to those obtained from quantum mechanical computations. Moreover, the geometric improvement takes place without downgrading the energetic agreement obtained from SIBFA. The systems considered are Cu(II) interacting with six water molecules, four ammonia or four imidazoles, and four water plus two formate anions. PMID- 14531051 TI - Intraprotein electrostatics derived from first principles: divide-and-conquer approaches for QM/MM calculations. AB - Two divide-and-conquer (DAQ) approaches for building multipole-based molecular electrostatic potentials of proteins are presented and evaluated for use in QM/MM calculations. One approach is a further development of the neutralization method of Bellido and Rullmann (J Comput Chem 1989, 10, 479-487) while the other is based on removing part of the electron density before performing the multipole expansion. Both methods create systems with integer charges without using charge renormalization. To determine their performance in terms of location of cuts and distance to QM region, the new DAQ approaches are tested in calculations of the proton affinity of N(zeta) of Lys55 in the inhibitor turkey ovomucoid third domain. Finally, the two methods are used to build a variety of MM regions, applied to calculations of the pK(a) of Lys55, and compared to other computational methodologies in which force field charges are employed. PMID- 14531052 TI - Ab initio quality properties for macromolecules using the ADMA approach. AB - We describe new developments of an earlier linear scaling algorithm for ab initio quality macromolecular property calculations based on the adjustable density matrix assembler (ADMA) approach. In this approach, a large molecule is divided into fuzzy fragments, for which quantum chemical calculations can easily be done using moderate-size "parent molecules" that contain all the local interactions within a selected distance. If greater accuracy is required, a larger distance is chosen. With the present extension of this approximation, properties of the large molecules, like the electron density, the electrostatic potential, dipole moments, partial charges, and the Hartree-Fock energy are calculated. The accuracy of the method is demonstrated with test cases of medium size by comparing the ADMA results with direct quantum chemical calculations. PMID- 14531053 TI - Tork: Conformational analysis method for molecules and complexes. AB - A conformational search method for organic molecules and bimolecular complexes is presented. The method, termed Tork, uses normal-mode analysis in bond-angle torsion coordinates and focuses on a key subset of torsional coordinates to identify natural molecular motions that lead the initial conformation to new energy minima. New conformations are generated via distortion along these modes and their pairwise combinations, followed by energy minimization. For complexes, special treatment is accorded to the six coordinates that specify the position and orientation of one molecule relative to the other. Tests described here show that Tork is highly efficient for cyclic, acyclic, and mixed single molecules, as well as for host-guest complexes. PMID- 14531055 TI - Bond energies for molecules, clusters, and deposit systems. AB - A new approach is suggested to the assignment of bond energies in molecules and clusters. It uses a shareholder principle for the redistribution of the shifts in atomic energies, which arise in a molecule, on the bonds. The scheme is directly suitable for semiempirical methods, where only one- and two-center terms occur. MSINDO calculations are performed to demonstrate the suitability of the approach for molecules and clusters. As an application the bonding in a deposit system is analyzed for the case of copper on magnesium oxide. It is found that copper atoms do not only bind to the preferred oxygen sites but also substantially to the magnesium sites. The copper-copper bonds are the strongest and will determine the structure of copper clusters on magnesium oxide surfaces. PMID- 14531054 TI - A point-charge force field for molecular mechanics simulations of proteins based on condensed-phase quantum mechanical calculations. AB - Molecular mechanics models have been applied extensively to study the dynamics of proteins and nucleic acids. Here we report the development of a third-generation point-charge all-atom force field for proteins. Following the earlier approach of Cornell et al., the charge set was obtained by fitting to the electrostatic potentials of dipeptides calculated using B3LYP/cc-pVTZ//HF/6-31G** quantum mechanical methods. The main-chain torsion parameters were obtained by fitting to the energy profiles of Ace-Ala-Nme and Ace-Gly-Nme di-peptides calculated using MP2/cc-pVTZ//HF/6-31G** quantum mechanical methods. All other parameters were taken from the existing AMBER data base. The major departure from previous force fields is that all quantum mechanical calculations were done in the condensed phase with continuum solvent models and an effective dielectric constant of epsilon = 4. We anticipate that this force field parameter set will address certain critical short comings of previous force fields in condensed-phase simulations of proteins. Initial tests on peptides demonstrated a high-degree of similarity between the calculated and the statistically measured Ramanchandran maps for both Ace-Gly-Nme and Ace-Ala-Nme di-peptides. Some highlights of our results include (1) well-preserved balance between the extended and helical region distributions, and (2) favorable type-II poly-proline helical region in agreement with recent experiments. Backward compatibility between the new and Cornell et al. charge sets, as judged by overall agreement between dipole moments, allows a smooth transition to the new force field in the area of ligand binding calculations. Test simulations on a large set of proteins are also discussed. PMID- 14531056 TI - Theoretical study of the internal elimination reactions of xanthate precursors. AB - The gas-phase internal elimination (E(i)) reaction of ethyl xanthate (CH(3)-CH(2) S-CS-O-CH(3)) has been investigated by means of Hartree-Fock, second-order Moller Plesset, and density functional theory (DFT) using the Becke three-parameter Lee Yang-Parr (B3LYP) functional and the modified Perdew-Wang one-parameter model for kinetics (MPW1K). Considerable differences between the ground- and transition state geometries and the calculated activation energies are observed from one approach to the other, which justifies first a careful calibration of the methods against the results of benchmark CCSD(T) calculations. Compared with these, DFT calculations along with the MPW1K functional are found to be an appropriate choice for describing the E(i) reaction of xanthate precursors. The precursor conformation and the transition states involved in the internal conversion of xanthate precursors of cyano derivatives of ethylene, and of cis- and trans stilbene, are then characterized in detail by means of this functional. PMID- 14531057 TI - Computing motif correlations in proteins. AB - Protein motifs, which are specific regions and conserved regions, are found by comparing multiple protein sequences. These conserved regions in general play an important role in protein functions and protein folds, for example, for their binding properties or enzymatic activities. The aim here is to find the existence correlations of protein motifs. The knowledge of protein motif/domain sharing should be important in shedding new light on the biologic functions of proteins and offering a basis in analyzing the evolution in the human genome or other genomes. The protein sequences used here are obtained from the PIR-NREF database and the protein motifs are retrieved from the PROSITE database. We apply data mining approach to discover the occurrence correlations of motif in protein sequences. The correlation of motifs mined can be used in evolution analyses and protein structure prediction. We discuss the latter, i.e., protein structure prediction in this study. The correlations mined are stored and maintained in a database system. The database is now available at http://bioinfo.csie.ncu.edu.tw/ProMotif/. PMID- 14531058 TI - Theoretical approach to the mechanism of reactions between halogen atoms and unsaturated hydrocarbons: the Cl + propene reaction. AB - The potential energy surface for the Cl + propene reaction was analyzed at the MP2 level using Pople's 6-31G(d,p) and 6-311+G(d,p), and Dunning's cc-pVDZ and aug-cc-pVDZ basis sets. Two different channels for the addition reaction leading to chloroalkyl radicals and five alternative channels for the abstraction reaction leading to C(3)H(5) (.) + HCl were explored. The corresponding energy profiles were computed at the QCISD(T)/aug-cc-pVDZ//MP2/aug-cc-pVDZ level of theory. Theoretical results suggest that the previously established mechanism consisting of (1) direct abstraction and (2) addition-elimination steps is instead made up of (1) addition through an intermediate and (2) two-step abstraction processes. No direct abstraction mechanism exists on the potential energy surface. The kinetic equations derived for the new mechanism are consistent with the pressure dependence experimentally observed for this reaction. PMID- 14531060 TI - An environmental pseudopotential approach to molecular interactions: Implementation in MOLPRO. AB - We present the implementation into the MOLPRO package of a model for the interaction of a central system with its surrounding environment. The properties of a target system enclosed by a noncovalently bound environment or solvent are modeled as those of a system embedded into the effective pseudopotential arising from the exact electrostatic Coulomb potential and the approximated exchange repulsion potential. For the latter we use the charge-density overlap model, which relates the exchange-repulsion interaction energy between two species with the overlap of their ground-state electron charge densities. The solutions of the modified Hartree-Fock equations for the target system are obtained self consistently. This way the exchange-induction effects arising from the converged electron-charge density of the embedded system are implicitly included. Inclusion of the correlation effects is provided by the use of post-Hartree-Fock and density-functional techniques available in the MOLPRO package. The computational and conceptual advantages provided by this approach are shown in the calculation of the dipole polarizabilities of halide and chalcogenide anions in different environments. PMID- 14531059 TI - Constructing irregular surfaces to enclose macromolecular complexes for mesoscale modeling using the discrete surface charge optimization (DISCO) algorithm. AB - Salt-mediated electrostatics interactions play an essential role in biomolecular structures and dynamics. Because macromolecular systems modeled at atomic resolution contain thousands of solute atoms, the electrostatic computations constitute an expensive part of the force and energy calculations. Implicit solvent models are one way to simplify the model and associated calculations, but they are generally used in combination with standard atomic models for the solute. To approximate electrostatics interactions in models on the polymer level (e.g., supercoiled DNA) that are simulated over long times (e.g., milliseconds) using Brownian dynamics, Beard and Schlick have developed the DiSCO (Discrete Surface Charge Optimization) algorithm. DiSCO represents a macromolecular complex by a few hundred discrete charges on a surface enclosing the system modeled by the Debye-Huckel (screened Coulombic) approximation to the Poisson-Boltzmann equation, and treats the salt solution as continuum solvation. DiSCO can represent the nucleosome core particle (>12,000 atoms), for example, by 353 discrete surface charges distributed on the surfaces of a large disk for the nucleosome core particle and a slender cylinder for the histone tail; the charges are optimized with respect to the Poisson-Boltzmann solution for the electric field, yielding a approximately 5.5% residual. Because regular surfaces enclosing macromolecules are not sufficiently general and may be suboptimal for certain systems, we develop a general method to construct irregular models tailored to the geometry of macromolecules. We also compare charge optimization based on both the electric field and electrostatic potential refinement. Results indicate that irregular surfaces can lead to a more accurate approximation (lower residuals), and the refinement in terms of the electric field is more robust. We also show that surface smoothing for irregular models is important, that the charge optimization (by the TNPACK minimizer) is efficient and does not depend on the initial assigned values, and that the residual is acceptable when the distance to the model surface is close to, or larger than, the Debye length. We illustrate applications of DiSCO's model-building procedure to chromatin folding and supercoiled DNA bound to Hin and Fis proteins. DiSCO is generally applicable to other interesting macromolecular systems for which mesoscale models are appropriate, to yield a resolution between the all-atom representative and the polymer level. PMID- 14531061 TI - A dynamic mean field theory for dissipative interacting many-electron systems: Markovian formalism and its implementation. AB - To demonstrate its applicability for realistic open systems, we apply the dynamic mean field quantum dissipative theory to simulate the photo-induced excitation and nonradiative decay of an embedded butadiene molecule. The Markovian approximation is adopted to further reduce the computational time, and the resulting Markovian formulation assumes a variation of Lindblad's semigroup form, which is shown to be numerically stable. In the calculation, all 22 valence electrons in the butadiene molecule are taken as the system and treated explicitly while the nuclei of the molecules are taken as the immediate bath of the system. It is observed that (1) various excitations decay differently, which leads to different peak widths in the absorption spectra; and (2) the temperature dependences of nonradiative decay rates are distinct for various excitations, which can be explained by the different electron-phonon couplings. PMID- 14531062 TI - Reassessment of methyl rotation barriers and conformations by correlated quantum chemistry methods. AB - Internal rotations of the methyl group in ortho-substituted and 2,6-disubstituted toluenes in their ground state have been investigated by means of various ab initio quantum chemistry methods. Computed barriers at the Hartree-Fock (HF) level using medium sized basis sets agreed reasonably with experimental results in the case of the studied ortho-substituted toluenes. However, this agreement worsens when using very large basis sets. Furthermore, the determination of the conformation and barriers of more weakly hindered methyl groups, that is, for 2,6 dihalogenotoluenes or toluene itself, necessitates high level correlated computations, because of a possible failure of HF calculations in this case. Density functional theory (DFT) techniques required, in several cases, much more extended basis sets than the post-HF Moller-Plesset perturbation (MP2, MP4) ones, to insure the convergence of the computed barriers. Non-negligible variations of the computed barriers when using different DFT functionals are observed for some systems. PMID- 14531063 TI - Effect of the N-terminal domain of the coat protein of potato virus X on the structure of viral particles. PMID- 14531064 TI - Lysozyme of the bivalve mollusk Unio pictorum. PMID- 14531065 TI - Interaction of the S100A4 (Mts1) protein with septins Sept2, Sept6, and Sept7 in vitro. PMID- 14531066 TI - Modifying effect of Mn(II) amino acid chelate on structural and functional properties of erythrocyte membranes in animals exposed to ionizing radiation. PMID- 14531068 TI - Study of the luminescence system of the soil enchytraeid Fridericia heliota (Annelida: Clitellata: Oligochaeta: Enchytraeidae). PMID- 14531067 TI - An adamantanone derivative that is an original modulator of redox processes in the cytochrome P-450 system can serve as an effective remedy against obliterating angiopathy of lower extremities. PMID- 14531069 TI - Three-dimensional reconstruction of the chromatin structure in the somatic nucleus of the infusorian Bursaria truncatella. PMID- 14531071 TI - The cholinesterase activity of the brain of northern Pacific flatfish of different species and genera of the family Pleuronectidae: substrate specificity. PMID- 14531070 TI - Phosphorylation of pea polypeptides induced by forskolin. PMID- 14531072 TI - Inhibition of the activity of recombinant HIV-1 integrase by derivatives of higher terpenoids. PMID- 14531073 TI - Study of the interaction between the Su(Hw) insulator and the binding sites for the GAF protein using the yellow gene regulatory system in Drosophila melanogaster. PMID- 14531074 TI - Tyrosine phenol-lyase: the role of the coenzyme-binding residue Ser-254 in catalysis. PMID- 14531075 TI - Interaction between zeste binding sites does not block isolated yellow enhancers in Drosophila melanogaster. PMID- 14531076 TI - Carotenoids are not required to provide protection of bacteriochlorophyll clusters against photooxidation in light-harvesting complexes of photosynthetic bacteria. PMID- 14531077 TI - Polyethylene-glycolated immunoliposomes specific for astrocytes. PMID- 14531078 TI - [Heavy metals in the soil of allotment gardens in industrialized post-flooded areas in the Opole region]]. AB - The results of studies concerning the levels of heavy metals (chromium, zinc, cadmium, nickel and lead) in soils of allotment gardens in post-flooded industrialized areas of the Opole Region, conducted during the period 2000-2001 was presented. Spectrophotometric method was applied for the determination of heavy metals in soils. The results of the study show that the levels of chromium, zinc and cadmium in the soils examined are constantly decreasing. However, an opposite phenomenon is observed with respect to copper and lead, where an increase was noted in the number of samples exceeding the maximum allowable, concentration of these metals during the second year of the study. This suggests that there exist local sources of contamination with the above-mentioned elements. PMID- 14531079 TI - [Changes in microbiological quality of kefir and yoghurt on the Warsaw market in the years 1995-2001]. AB - The microbiological quality of kefir (61 samples) and yoghurt (92 samples) purchased in retail network in Warsaw in the years 1995 and 2001 has been examined. The total number of bacteria in at least 90% of yoghurt and 73% of kefir was in the range 10(7)-10(9) c.f.u. in 1g. The dominating group of bacteria in kefir were mesophilic lactic acid streptococci and in yoghurt (analysed in the year 2001)--S. thermophilus. The number of L. delbrueckii subsp. bulgaricus in 40% of samples was < 10(7)-in 1 g. The increment of microbiological quality of the examined fermented milks has been observed. More than 86% of kefir and 97% of yoghurt analysed in 2001 were free from coliform bacteria, most of them free from B. cereus, moulds and yoghurt--also from yeasts. About 48% of kefir samples didn't fulfilled the requirements of FAO/WHO requirements concerning the number of yeasts. PMID- 14531080 TI - [Modification and validation of the spectrophotometric method of trace amounts of antimony determination in different types of mineral water]. AB - In order to improve limit of determination, a modification of the analytical procedure for spectrophotometric determination of antimony in water recommended by Polish Standards has been proposed. The modified procedure was validated on the basis of the analytical studies of samples of deionised water enriched in different amounts of the analyte, and its precision, limit of determination, sensitivity, accuracy and linearity range were established. A correction describing the effect of the matrix, determined on the basis of analyses of four types of mineral water differing in the degree of total mineralisation and chemical composition, and allowing the use of the procedure proposed for determination of antimony at the level of < or = 0.005 mg/dm3, has been derived. PMID- 14531081 TI - [Silicon forms and its role in infant food]. AB - This paper presents an attempt to assess the total content of silicon and its potentially bioavailable forms to the baby body in milk nutrient and baby foods. The measurements of the total and bioavailable silicon were conducted with the use of two analytical methods. The method of atomic absorption spectroscopy (AAS) was used for the determination of total silicon content. Bioavailable silicon was determined by means of visible light spectroscopy (VIS) with ammonium molybdate. In the group milk products the largest amount of total silicon was found in Mlekovita milk -0.54 mg/l, and Humana 2 milk formula contained the largest amount of ionised silicon-0.24 mg/l. In baby foods manufactured by Gerber the total amount of silicon was high and the average content stood at 27 mg/kg in dry matter. The largest amount of total silicon was found in Vegetables and Ham-48.19 mg/kg in dry matter, as well as bioavailable silicon--4.13 mg/kg in dry matter. Compare level of silicon in milk nutrient and baby foods to silicon content in human milk 0.05 mg/l, it was found that large excess of silicon in this foodstuffs. PMID- 14531082 TI - [Fat and fatty acid content in selected powdered ready to eat dishes]. AB - Fat content as well as fatty acids composition in 16 powdered ready to eat dishes from the market in Warsaw area in the year of 2001 has been analysed. We have found, that fat content in these products was from 5.2 to 24.1%. Saturated fatty acids were dominant in the fat derived of these products and also trans isomers unsaturated fatty acids, know from recent finding as having negative nutritional effect. PMID- 14531083 TI - [Polyethylene terephthalate (PET)--health aspects and food packaging application]. AB - Polyethylene terephthalate (PET) due to its physicochemical properties, especially regidity and glass-like transparency is widely used as food packaging material. The relevant legislation states that substances may not migrate from food contacting materials in quantities that may cause undesirable changes in organoleptic properties of food coming into contact with such material. The lists of substances authorized for food contact plastic materials and requirements for the final product were established. The requirements concern global migration limits (60 mg/kg or 10 mg/dm2) and specific migration limits (SML) set for substances which, when migrate into food in grater quantities may cause risk for human health. For the products manufactured from PET the specific migration limits were set for terephthalic acid (7.5 mg/kg), for isophthalic acid (5 mg/kg), for isophthalic acid dimethyl ester (0.05 mg/kg) and for ethylene and diethylene glycol (30 mg/kg). PET may undergo thermal degradation resulting in formation of acetaldehyde, which may influence organoleptic characteristics of packaged foods changing taste and smell. PMID- 14531084 TI - [Evaluation of students' nutrition in Poland based on the data for the year 2001 gathered by the Sanitary-Epidemiological Stations]. AB - Having used reports from sanitary-epidemiological stations, a situation of schools supplementary nutrition in all types of schools in Poland in the year 2001 has been analyzed. It has been stated, that a hot meal is consumed by 16.1% (1,090,535) of pupils. It has been reported, that schools serving meals probably met demand, but meals were not provided in 65% of schools. In these institutions, creating possibility of serving meals appears to be the most important task on the way to improve pupils nutrition in schools. In 60% of schools such conditions need to be established from the foundation, while in 5% of schools it is enough to make use of existent potential. It has been emphasized, that a pupil of every school should have possibility to have a drink (today it is possible in 32% of schools) as well as to eat a sandwich. Milk products were consumed in 4% of schools and it concerned 2% of pupils. In 40% of schools a pupil could purchase in a grocery articles of consumption, mainly confectionery articles and non-milk drinks. However, pupils should be encouraged and stimulated to purchase in groceries products reinforcing rational nutrition. PMID- 14531085 TI - [Nutrition quality of daily food intake of the Warsaw population in the years 1993-2001. Warsaw Pol-MONICA bis project]. AB - The aim of the study was to evaluate changes in dietary habits and the realization degree of the recommended dietary allowances of vitamins and minerals intake in the diet of the Warsaw inhabitants in the 8-year period. In years 1993 and 2001 dietary habits where assessed according to 1485 and 836 randomly selected 35-64 year old men and women. Over the observation period substantial changes in food consumption pattern were noticed: the percentage of energy from fat decreased from 39.9% to 37.8% in men and from 38.2% to 35.9% in women, the percentage of energy from saturated fatty acids decreased from 14.0% to 12.4% in men and from 13.5% to 11.9% in women, the percentage from carbohydrates increased from 46.0% to 47.5% and from 47.5% to 49.6% and dietary fibre intake from 21.5 g to 22.8 g and from 15.7 g to 17.5 g respectively. In 2001 in comparison with 1993 intakes of vitamins C and B2, calcium, magnesium, potassium and iron were higher in both genders, vitamin B1 in men and E in women. Despite those positive changes still in year 2001 fibre, vitamins B1 and B2 and analyzed minerals intake was on too low level. The highest deficiency has been noticed in calcium intake in both genders and iron in women-the realization degree of recommended intake has stayed on 60-67% level. PMID- 14531086 TI - [Characteristics of nourishment of youth starting studies at the Academy of Physical Education in Cracow]. AB - The purpose of the research has been to analyse the preferences and dietary habits of the first-year-students of Physical Academy. The research was carried out on 115 female and 110 male students. The research has shown a great deal of defective nourishment and the necessity of monitoring nourishing ways of the students. PMID- 14531087 TI - [The frequency of consuming first and second breakfasts among children in rural areas]. AB - The investigation included 1,154 schoolchildren at the age of 7-15 who come from rural areas. With the aid of an interview information was obtained concerning the consumption of first breakfast, beverage and second breakfast on the day before the investigation. Almost one fifth of the interviewed children did not eat first breakfast on the day of the investigation. There was a greater percentage of schoolchildren who did not drink any beverage during their stay at school, and children who did not eat regularly the meals under investigation. The interviewed girls used to consume meals more regularly than boys. Girls ate their second breakfast more often, however, they had beverages more rarely during their stay at school. Dependence was found between the level of the education of mother and the frequency of the consumption of second breakfast and beverages at school as well as the regularity of the consumption of meals among the interviewed schoolchildren. A more regular consumption was associated with better grades achieved by the investigated schoolchildren. PMID- 14531089 TI - [Expectations regarding nutrition information on food product packages]. AB - Food labeling was originally developed to help consumers make the right decision in choosing at food products. It (food labeling) is also helpful in consumer's education. The aim of this study was to analyze the consumers' preferences concerning foods' labeling products. The type and the form of the information and of course the information itself were analyzed. The study was carried out in the form of a questionnaire among 295 people who were the consumers of supermarkets. The only criteria for the chosen subjects were that they agree to take part in the study and that they are the heads of their families. It came out that 54% of the subjects accepted the current information about food products. More than 23% said that they didn't understand the information provided by the food labels. 66% said that food labels should have educational information concerning the role of an ingredient in health and nutrition. Everyone wanted "negative" information, for example: "this product is not recommended for..."). More than 87% stated that they want information like "this product is recommended for...". (That kind of preference doesn't go along with the current law concerning food products). The best motivation for using food labels is sickness or illness. 89% of the subjects said that the information about the food products and the nutritional value are more important than the information about the producer. Informations concerning the nutritional values in 100 g and in 1 portion of a product were easiest to understand for 71% of the subjects. The results of this study show that food labeling is very important form of consumers education. PMID- 14531088 TI - [Nutrition behavior of poor families]. AB - The aim of this study was analysis of nutrition behaviour of families with low income living in village and town. The interviews concerning socio-economical conditions, actual nutrition behaviour and selected health indicator of respondents were carried in 13 rural and 14 Warsaw families. Respondents were for the most part women 34-44 years old with elementary education. Rural families were mainly numerous families, half of Warsaw families consists of mother lonely bringing children up. The main sources of living were allowances from social welfare and rents. One can observe in both kinds of families pro-healthy nutrition behavior: daily drinking milk by children, frequent consumption of poultry, using margarine and vegetables oil, and in rural families eating breakfast and several meals everyday. Negative aspects of nutrition were: small consumption of vegetables and fruits, fishes, and in town also dairy products and insufficient number of daily meals. Half of respondents both in village and town has proper BMI index. One can observe in rural families more tendency toward healthy nutrition behavior than in Warsaw families. Modeling healthy nutrition behavior in poor families (especially in town) can be realised by centers of social welfare. PMID- 14531090 TI - Highlighting need to avoid PEI malfunctions. PMID- 14531091 TI - Securing best development of healthcare facilities. PMID- 14531092 TI - Audiovisual technology demonstrates huge merit. AB - The use of state-of-the-art audiovisual technology is bringing enormous practical benefits to the health sector at large. Quadrant Visual Solutions describes the experience of Darent Valley Hospital which installed new AV systems in June 2003. PMID- 14531093 TI - Controlling legionella without damaging evaporative condensers. PMID- 14531094 TI - Improving risk management systematically. AB - The management of risk is an ever more prevalent issue amongst healthcare professionals due to key changes in recent years driving visibility and transparency. Medical gases constitute an important area of risk--the gases are used, administered and handled by many different people and the chief executive is ultimately responsible. Such risks can be managed down by a hospital according to the letter and spirit of HTM 2022, EN737 and NMSR. The biggest failing of the current system where hospitals take it upon themselves to ensure compliance, is that typically little training is actually performed, there is almost no monitoring, and often people on the front line are unaware of the risks that they are running. A systematic response to managing risk is the only way forward, and a key decision a hospital needs to make is whether to persist towards this goal itself, or to involve a technical and clinical supplier. BOC Medical is in a key position to be of value. PMID- 14531095 TI - Patient monitoring: the big picture. AB - Increased provision of patient monitoring equipment can assist in the process of raising standards of care. The range of such equipment is extensive, and estates professionals including planners, engineers and facilities managers need to be aware of what items are appropriate to different clinical areas and the sort of back-up support the equipment will require. This article is formed from a report of an independent multi-disciplinary working group which reviewed patient monitoring technology and looked at factors involved in purchasing monitoring equipment. PMID- 14531096 TI - Concept overcomes 'blight of the bland'. PMID- 14531097 TI - Advanced communication technology vital. PMID- 14531098 TI - Making a pipe dream come true. PMID- 14531099 TI - Astonishing indifference to deaths due to botched ritual circumcision. PMID- 14531100 TI - Hormone replacement therapy. PMID- 14531101 TI - Community service in rural South Africa. PMID- 14531102 TI - Radiologists 'walk' in contrast conflict. PMID- 14531103 TI - Much ado about ethics at UCT clinic. PMID- 14531104 TI - Behave + pay costs, HPCSA tells guilty Drs. PMID- 14531105 TI - Fine needle finesse. PMID- 14531106 TI - The inevitability of antiretrovirals. PMID- 14531107 TI - Impact of HIV on children--UCT report. PMID- 14531108 TI - Campaign to end polio by 2005. PMID- 14531109 TI - Gauteng health calls for retrieval of unused medicines and drugs. PMID- 14531110 TI - The Traditional Health Practitioners Bill, 2002. PMID- 14531111 TI - Medical organisations. PMID- 14531112 TI - Ethical issues. Part 2. PMID- 14531113 TI - Phytosterols--a new dietary aid for the treatment of hypercholesterolaemia. PMID- 14531114 TI - Savings from generic drug substitution in South Africa--an arithmetical view. PMID- 14531115 TI - Chronic conditions--the new health challenge. PMID- 14531116 TI - Value can be added to the health care system. PMID- 14531117 TI - Endovascular management of intracranial aneurysms--current status in South Africa. PMID- 14531118 TI - World pandemic of obesity--any hope of its being controlled? PMID- 14531119 TI - Candida famata central nervous system infection. PMID- 14531120 TI - Exclusive breast-feeding--a pipe dream? PMID- 14531121 TI - Missed opportunities for vaccination in health facilities in Swaziland. AB - OBJECTIVES: To determine whether potential exists to increase vaccination coverage in Swaziland by reducing missed opportunities. DESIGN: The standard World Health Organisation Expanded Programme on Immunisation (WHO EPI) protocol for assessing missed opportunities for vaccination was used to conduct this study. It involved client exit interviews and review of health cards. SETTING: Selected variety of health facilities in Swaziland. SUBJECTS: Children less than 2 years of age and women of child-bearing age exiting each facility. OUTCOME MEASURES: Children and women eligible for vaccination exiting sampled health facilities. RESULTS: Fifty-four per cent of eligible children less than 2 years of age were missed for vaccination. This constitutes 26% of all children less than 2 years old leaving the facilities studied. Almost 100% of eligible women of childbearing age were missed for vaccination, constituting 88% of women leaving the study facilities. The distribution of the proportion of missed opportunities varied considerably between regions and health facility types. Missed opportunities occurred more frequently among those children requiring the first dose of all antigens and this may be linked to the high proportion of children missed for vaccination who did not possess a health card. Missed opportunities were more likely to occur in facilities providing integrated services. CONCLUSIONS: The frequent attendance at health facilities of the target group presents a valuable opportunity to increase vaccination coverage through avoidance of missed opportunities. All regions need to set vaccination coverage targets and develop plans to increase coverage, which should include strategies to ensure that all health workers routinely screen all clients for eligibility and vaccinate as required. PMID- 14531122 TI - Back injuries in young fast bowlers--a radiological investigation of the healing of spondylolysis and pedicle sclerosis. AB - OBJECTIVE: To demonstrate the efficacy of various radiological diagnostic modalities in assessing lower back pain in young fast bowlers. METHODS: Ten cricketers who presented to either a physiotherapist or a doctor with suspected spondylolysis underwent an X-ray, a single photon emission computed tomography (SPECT) bone scan and a computed tomography (CT) scan to assess the severity of the injury. Three and 12 months after the initial CT scan, second and third CT scans were performed in order to assess whether healing had taken place. After the initial radiological investigation the subjects diagnosed with spondylolysis or pedicle sclerosis underwent prescribed intervention and rehabilitation which included physiotherapy modalities, postural correction, and specific individually graded flexibility, stabilisation, strengthening and cardiovascular programmes. RESULTS: Radiographs were normal in 8 subjects, while 2 had evidence of sclerosis. The isotope scan showed increased uptake in all of the subjects. The CT scans showed no fracture (N = 3), partial fractures (N = 3), complete fractures (N = 2) and old fractures bilaterally (N = 2). When the follow-up CT scan was carried out at 3 months, 1 of the subjects had developed a partial fracture of the left pars interarticularis on the inferior border, which showed complete union when CT scanned at 12 months. At 3 months the partial and complete fractures showed progressive healing in 2 subjects, with complete healing in all the other cases. Complete healing was achieved in all subjects at 12 months, with the exception of 1 subject who showed near-complete union, with a small area of fibrous union on the inferior border and 2 old bilateral fractures that remained un-united. RESULTS: From the results it is evident that when a young fast bowler presents with backache after bowling, it would be appropriate to do an X-ray, a bone scan and a CT scan to make the diagnosis. Discontinuing the fast bowling and following an active rehabilitation programme should result in spontaneous resolution and healing of the fractures. If it is not detected early a fibrous or non-union fracture could result. PMID- 14531123 TI - HIV-exposed infants with acute respiratory failure secondary to acute lower respiratory infections managed with and without mechanical ventilation. AB - OBJECTIVES: The decision to provide mechanical ventilation (intermittent positive pressure ventilation (IPPV)) to HIV-exposed infants in resource-poor settings has remained difficult owing to problems in confirming HIV infection and the lack of data on outcome. We evaluated the predictive value of the HIV antibody test in confirming infection in infants requiring mechanical ventilation for acute lower respiratory infections (ALRIs), and compared the outcome for children denied access with the outcome for similar subjects who were ventilated. SETTING AND DESIGN: This investigative study was conducted over a 12-month period at the paediatric intensive care unit (PICU) at King Edward VIII Hospital (KEH) in Durban, and at Ngwelezana Hospital in northern KwaZulu-Natal. SUBJECTS: HIV exposed patients with acute respiratory failure (ARF) secondary to ALRI entering the PICU at KEH were enrolled into the IPPV arm, while similar children who were refused such care at Ngwelezana Hospital were admitted into the non-IPPV arm. Standardised protocols for entry and management of enrolled subjects were utilised. OUTCOME MEASURES: HIV DNA polymerase chain reaction (PCR) testing was performed to establish HIV status. Clinical and laboratory parameters were correlated with HIV status to determine predictors of infection and outcome (survival to discharge). RESULTS: One hundred and sixteen HIV-exposed infants were enrolled, 49 into the IPPV arm and 67 into the non-IPPV arm. The median age of both groups was 3.0 months (0.5-11 months), and the male/female ratio and proportion of infants under 3 months of age were similar in both groups. The predictive values of the HIV antibody test in determining HIV infection in the IPPV and non-IPPV arms were 87.8% and 85.0% respectively. Splenomegaly and a serum globulin of > 35 g/l increased the likelihood of being HIV PCR-positive (p = 0.006 and p = 0.04 respectively). Survival to discharge rates for HIV-infected children in the IPPV and non-IPPV arms were 41.9% and 24.6% respectively (p = 0.08). Age less than 3 months (p = 0.04) and very severe pneumonia (p = 0.007) were the only indicators of poor outcome. CONCLUSION: Mechanical ventilation provided little benefit in HIV-infected children with ARF from ALRI. An HIV antibody test in infants with ALRI and ARF is highly suggestive of HIV infection. Splenomegaly and a serum globulin of greater than 35 g/l were the only useful markers in identifying HIV infection. PMID- 14531124 TI - Occupational exposure to blood in medical students. AB - OBJECTIVE: To determine the extent of occupational exposure to blood in medical students, details of the circumstances surrounding the incidents and the subsequent experiences of the student. DESIGN: Prospective cohort study. SETTING: Tygerberg Hospital, the Health Sciences Faculty of the University of Stellenbosch during a 15-week period from 4 February to 19 May 2002. SUBJECTS: One hundred and thirty-six student interns (SIs), i.e. final-year medical students. METHOD: All Sis received a questionnaire and a letter motivating them to participate in the study and explaining the procedure. Regular class meetings enabled continuous motivation and ongoing update. In the case of an incident during the 15-week period, the SI filled in the form and placed it in a sealed drop-off box. OUTCOME MEASURES: Specific focus on the preceding events and the situation in which the incidents occurred (department, time of day, procedure performed, and whether the student was on call), exposure to HIV (patient's retroviral status), use of post exposure prophylaxis (PEP) (whether used, when initiated), and the consequences of the exposure (emotional, on sexual behaviour during the window period, and on career choice). RESULTS: During the 15-week period, 19 incidents were reported; the majority occurred while students were on call, almost half occurred after hours, and a disproportionate number occurred in three departments. CONCLUSIONS: Occupational blood exposure is a very real problem and poses a significant risk. SI suggestions should be considered in improving the prevention and management of such incidents. PMID- 14531125 TI - [Simulators in medical education spare patients from unskilled personnel]. PMID- 14531126 TI - [Treatment with levodopa can affect latent vitamin B 12 and folic acid deficiency. Patients with Parkinson disease runt the risk of elevated homocysteine levels]. AB - There is a well-known interaction between vitamin B12, folate, and homocysteine. More unknown is the fact that this interaction might be affected by long-term treatment with levo-dopa in patients with Parkinson's disease. An increase in homocysteine levels and tissue deficiency of vitamin B12 and folate may occur. The responsible doctor should be liberal in checking vitamin B12 and folate status and supplement with appropriate vitamins when needed. PMID- 14531127 TI - [High-energy microwave therapy with benign prostatic hyperplasia. A good and safe therapeutic choice--for both the patient and the health care]. PMID- 14531128 TI - [Treatment of patients with liver metastases from colorectal cancer. More patients can benefit from the treatment!]. AB - Liver resection for liver metastases goes with 30-40% five years survival. It is estimated that 10% of patients with liver metastases can be subjected to liver resection. In the Vastra Gotaland region, this number is not achieved. In the present material of 147 patients, the postoperative mortality was 2.7%. Five years survival was 33%. Preoperative chemotherapy and preoperative porta embolisation have extended the indication for liver resection. Reresection after recurrence limited to the liver should be considered in selected cases. Ablative measures are under development and should be evaluated in clinical trials. New chemotherapeutic drugs (oxaliplatin, irinotecan) with improved recurrence rate but with limited gain in survival might have an impact as adjuvant therapy. A surgeon with liver surgery competence should see patients with a presumably resectable cancer. PMID- 14531129 TI - [C-reactive protein potential risk factor for cardiovascular disease]. AB - C-reactive protein (CRP) is a protein produced mainly in the liver upon stimulation by interleukin-6. CRP is part of the acute phase response to inflammation and plasma concentrations rise 100-1000 times in patients with for example bacterial infections. High sensitive methods have made it possible to measure and compare low CRP levels in plasma. Prospective studies have shown that increased but comparatively low plasma CRP concentrations are associated with future coronary heart disease (CHD) in healthy individuals and in patients with known CHD. Further support for an important role for CRP comes from in vitro studies that have suggested that the CRP protein is intimately and actively involved in the atherosclerotic process. This has stimulated the interest for clinical evaluation and treatment of increased plasma CRP levels. PMID- 14531130 TI - [Learning bronchoscopy in simulator improved dexterity rather than judgement]. PMID- 14531131 TI - [Alphonse Daudet--a syphilis victim: "There are long days when nothing but the suffering is alive inside me"]. PMID- 14531132 TI - [Capsulotomy in obsessive syndromes: medical matter-of-fact arguments or a forced neglect of the reality?]. PMID- 14531133 TI - [Final reply about snuff: increased awareness in the target]. PMID- 14531134 TI - [Final reply about the examination for interns: better to admit a mistake]. PMID- 14531135 TI - [Terry Fox--a Canadian national hero]. PMID- 14531136 TI - [Tuberculosis, love and death!]. PMID- 14531137 TI - [The good clinical stay]. PMID- 14531138 TI - [Research and the individual physician]. PMID- 14531139 TI - [Between science and practice. Science in practice--a theoretical perspective on learning]. PMID- 14531140 TI - [The interactive lecture. A simple form of student-activating learning]. AB - INTRODUCTION: Activation of students in lectures to enhance learning by means of questions to be answered in buzz groups has been described in pedagogic handbooks and articles. We tried the concept in three lectures in biochemistry in order to evaluate use of time, training requirements of the lecturer, and method acceptance by students. MATERIAL AND METHODS: The experiment was carried out with a group of 87 medical students from a 4th semester course in biochemistry. Evaluation was made by direct observation and analysis of quantitative and qualitative data from a student questionnaire. RESULTS: Buzz groups and questions took less time than anticipated and not more than ten minutes of the lecture time. The lecturer needed supervision from a colleague to function well. Acceptance of the procedure was high among the students. Qualitative data indicate that students used more time for self-studies and moved towards deep learning. DISCUSSION: We conclude that interactive lecture could be implemented without major problems in lecture based educational programmes and that it is useful for the learning of the students. PMID- 14531141 TI - [Medical students' clinical stay]. AB - In this article, central theories of learning in practice are discussed in relation to medical education, especially the first clinical stay. In the light of a project in the ward, the article attempts to demonstrate the importance of this stay, which is vital to students' development of a professional identity. In the theories of learning in practice, it is stressed that learning and development of a professional identity are inextricably linked. The article discusses what impact clinical stays have on students. It is stressed that it is necessary to create a structure for ward-based learning that will incorporate students into the daily working routine. PMID- 14531142 TI - [Learning clinical skills]. AB - INTRODUCTION: The purpose of this study was to analyse the learning environment of medical students in a medical ward and on the basis of this study to make specific suggestions as to how the students' clinical stay in a ward can be optimized. These suggestions were implemented and the effect evaluated. MATERIAL AND METHODS: This is a qualitative study. Through observations and interviews with students and staff in the ward the learning environment was analysed. After that an evaluation of the implemented project was carried out in a questionnaire to the ward and in interviews with the persons involved in the project. RESULTS: The result of the study concerning analysis and improvement of the students' stay can briefly be summarized as follows: The students' position in the unit can be characterized as that of an observer rather than as an active working participant in the ward. With this there is a risk that the medical student's trainee stay in a ward will turn into a "work-experience placement", the primary purpose of which will be for the medical student just to dip into the profession. This has consequences for the students' learning process in securing that the students acquire the necessary practical qualifications and acquire essential personal qualifications, such as self-dependence and abilities in decision-making and working under pressure. DISCUSSION: The study shows that doctors and students agree on the aim of the students' trainee stay: To involve the students in a working partnership in order to obtain the best learning experience. The study concludes that it is difficult to change well-established routines despite good intentions of all parties involved. Therefore, the study rises the question on how new routines are established in the educational culture of medicine. PMID- 14531143 TI - [Learning of house officers: from studies to practice]. PMID- 14531144 TI - [Educational needs of physicians and nurses working in palliative care]. AB - INTRODUCTION: The WHO definitions of palliative care have been adopted in Denmark and implemented in the national guidelines. The aim of this study was to assess the educational needs of doctors and nurses in palliative care in a Danish county hospital and the related primary care sector. MATERIALS AND METHODS: A self administered questionnaire was mailed to an anonymous cross-section of hospital nurses and doctors, home care nurses and GPs in the related primary care sector. The questions concerned the type of education obtained in the area and priorities for education in communication, supervision, pain and other typical symptoms in the terminally ill, as well as bereavement and grief therapy. The responses were compared by chi 2 statistics. RESULTS: 347 (76%) of those surveyed responded to the questionnaire. 81% of them were actually caring for a terminally ill or dying patient, and another 13% had done so within the past six months. The doctors, especially the hospital doctors, had generally received less education in the area surveyed than the nurses had and generally prioritized education lower than the nurses did; 44% of the respondents agreed completely or partially that they found it difficult to get up-to-date information on symptom control in palliative care. DISCUSSION: The findings indicate a profound need for education in palliative care. Doctors reported having received less education in this area than nurses had, and they also perceived a lower priority for education than nurses did. Particularly worrisome is the level of education of hospital doctors. PMID- 14531145 TI - [Physicians in training in general practice want mentors]. AB - INTRODUCTION: The new specialist training in family medicine in Denmark calls for increased educational time spent in general practice. The aim of this study was to describe the factors affecting the educational relationship between trainer and trainee, as seen from the perspective of the trainees. MATERIALS AND METHODS: The participants in the study were seven of eight trainees recently given the position of specialist training for general practice in a Danish county. We conducted repeated focus group interviews and individual interviews over a period of nine months. Our method was based on principles of ethnographic interviews. The data were analysed in the style of Kirsti Malterud. An English model of mentoring was used to establish a theoretical perspective on the data. RESULTS: Essential to the educational relationship between trainee and trainer is that the trainee feel welcome and that the initial interview succeed. It is also essential that the tutor be readily available, but trainees have varying need for supervision, which must be conducted with regard to each trainee's learning style and clinical competency. It is also essential that a dialogue of equals be established. The trainee is in a learning situation and needs positive feedback. When the educational relationship does not succeed, it is often due to the trainers' and trainees' differing expectations of the relationship. DISCUSSION: In specialist training for family medicine, it is taken for granted that the trainer functions as a mentor. The tutor is supposed not only to supervise the trainee in clinical work but also to create a framework in which the trainee can reflect on his or her personal and professional life. Trainees want mentors, but such a relationship does not automatically succeed. Success in mentoring in general practice depends on trainers' perception of their role as tutors, as well as, perhaps, on Danish doctors' culturally instilled professional values. PMID- 14531146 TI - [Research training for all physicians--also outside university hospitals. Evaluation of a course in basic research methods]. AB - INTRODUCTION: General practitioners are often interested in doing research but are hampered by lack of time and research training. Interpreting the results of others can also be difficult. For this reason a course in basic research methods for GPs was started in Sweden in 1989. It was originally aimed at GPs but was later extended to hospital physicians as well. The course is given regionally and at present is held in six different locations in southern Sweden as well as in Hillerod, Denmark. The aim of this study is to evaluate the course as part of a research project to recommend changes to the courses in accordance with the course evaluations. MATERIALS AND METHODS: The course consists of theory (lectures/seminars six hours a month) and practice (project work) over a period of 18 months. Questionnaires were mailed to the 112 physicians who, starting in 1997 and 1999, completed the course. Eighty-five percent responded to the questionnaire, which asked if the course goals were relevant and if they thought they had achieved them. RESULTS: The most frequent reasons given for attending the course were a desire to learn how to read scientific articles critically and how to carry out one's own research projects. Two thirds of the participants thought that the theoretical lectures and project work had supplemented each other well. Most of the participants thought that the goals were very relevant but fewer--between 57% and 77%--felt those goals had been achieved. DISCUSSION: The course provides important empirical information with regard to future specialist training in Denmark. The plan is for all physicians to have research training--amounting to a total of 60 days--with lectures in research methodology and tutored research projects. The Danish College of General Practitioners has planned research training in accordance with the experience from these courses. PMID- 14531147 TI - [Drug information leaflets confuse patients]. PMID- 14531148 TI - [The National Serum Institute's double role as expert adviser and test producer- the history of Mycoplasma genitalium]. PMID- 14531149 TI - [Questions to the Hearing Aid Center]. PMID- 14531150 TI - [Andropause--does it exist?]. PMID- 14531151 TI - [Retrospective studies and archiving of medical journals]. PMID- 14531152 TI - [Is the cognitive test suitable?]. PMID- 14531153 TI - [The public health portal for physicians--a double cost for the state?]. PMID- 14531154 TI - [Health, risk conditions and disease--thoughts about the "Health Profile" by the pharmaceutical society]. PMID- 14531155 TI - [A modern, rational health care--or physicians being wound up]. PMID- 14531156 TI - Respiratory psychophysiology and the modification of breathing behavior. PMID- 14531157 TI - Effects of left amygdala lesions on respiration, skin conductance, heart rate, anxiety, and activity of the right amygdala during anticipation of negative stimulus. AB - The present study reports the effects of lesions in the left amygdala on anxiety, respiration, skin conductance, heart rate, and electrical potentials in the right amygdala in two patients. Trait and anticipatory-state anxiety were measured before and after left amygdala resection to control medically intractable epilepsy in the patients. Lesions in the left amygdala resulted in decreases of trait and state anxiety, respiratory rate, and activity in the right amygdala in both patients; one patient also showed notable decreases in skin conductance and heart rate. The study also reports that activities in the right amygdala before the lesion were not observed after the lesion. We suggest that the activity of the right amygdala is dominantly activated in anxiety and anxiety-related physiological responses but needs excitatory inputs from the left amygdala. PMID- 14531158 TI - The conditioning of dyspneic suffocation fear. Effects of carbon dioxide concentration on behavioral freezing and analgesia. AB - Previous studies in our laboratory have shown that a single exposure to 100% carbon dioxide (CO2) can serve as an effective unconditioned stimulus (US) in a Pavlovian aversive-context conditioning paradigm in rats. Although the US exposure parameters employed in the initial studies were sufficient for producing a context-specific enhancement of behavioral freezing and analgesia, it had yet to be determined whether variations of these CO2 conditioning procedures would produce other conditioning effects. Thus, the purpose of the following experiment was to investigate the intensity of the US on the conditioned response (CR). The findings confirm that variations in CO2 concentrations produce changes in the CR that are consistent with principles of Pavlovian conditioning. The findings lend additional support to the tenability of a dyspneic suffocation fear theory of panic disorder, a theory that postulates that at least one type of panic attack could be a consequence of Pavlovian conditioning. PMID- 14531159 TI - Behavioral influences and physiological indices of ventilatory control in subjects with idiopathic hyperventilation. AB - Idiopathic hyperventilation has been defined as a respiratory-related psychophysiological complaint. This study attempted to clarify relationships between psychological and physiological variables in this condition. Participants demonstrated increased anxiety, depression, and symptoms consistent with hyperventilation. This was associated with a reduced peripheral chemosensitivity (isocapnic hypoxic rebreathe; -0.84 +/- 0.5 min-1.%O2(-1)), which was normalized with experimentally increased pCO2. Resting CO2 sensitivity was close to normal (2.1 +/- 1.0 min-1.mmHg-1). Breath hold time was significantly reduced versus controls (20.4 s +/- 12 s vs. 63 s +/- 31 s), and resting PETCO2 was correlated with the anxiety score. Also, the ventilatory response to moderate intensity exercise was augmented (vs. controls). The normalcy of pulmonary and chemoreceptor responses suggests that psychological factors may initiate this hyperventilation, which may become a conditioned response with an increased drive to breathe. PMID- 14531160 TI - Respiratory variability and psychological well-being in schoolchildren. AB - Among the relations between respiration and psychological state, associations with respiratory variability have been contradictory. In this study, respiration was measured noninvasively in 162 children with a mean age of 11 years (from 9 to 13). They completed a battery of psychological tests. Structural Equation Modeling (SEM or LISREL) revealed a model that fit the data well (chi 2 = 88.201, df = 79, p = .224). In this model, respiratory variability was positively related to anger-in and negatively to negative fear of failure and neurotic complaints. Respiration rate was positively related to positive fear of failure, and duty cycle was positively related to the latent variable of negative affect. Variability in resting time components of respiration was higher among children with less fear of failure and fewer complaints. Therefore, respiratory variability need not necessarily be a sign of psychological dysfunctions, and interventions should not always impose a fixed breathing pattern. PMID- 14531161 TI - The lifeShirt. An advanced system for ambulatory measurement of respiratory and cardiac function. AB - An accurate ambulatory breathing monitor is needed to observe acute respiratory changes in patients with medical or psychological disorders outside the clinic (e.g., hyperventilation during panic or apneas during sleep). Significant limitations of existing monitors are size, troublesome operation, and difficulty holding chest and abdomen bands in place during 24-hour recordings. Recently, a garment has been developed with embedded inductive plethysmography sensors for continuous ambulatory monitoring of respiration, heart activity, inductive cardiography, motility, postural changes, and other functions. The signals are displayed and stored on a handheld computer (Visor), and then analyzed offline, extracting more than 40 clinical parameters relating to cardiorespiratory function (e.g., heart rate, respiratory sinus arrhythmia, tidal volume, stroke volume, pre-ejection period, apnea-hypopnea index, thoraco-abdominal coordination, sighing). The device also serves as an electronic diary of symptoms, moods, and activities. This advanced system may open a new era in ambulatory monitoring for clinical practice and scientific research. PMID- 14531162 TI - Clinical applications of breathing regulation. Beyond anxiety management. AB - Breathing training is widely used as an aid in reducing anxiety states, but several other applications also show promise. This article reviews evidence that normalizing breathing patterns may offer help in some cases of essential hypertension, angina, functional chest disorder, chronic obstructive pulmonary disease (COPD), and cardiac rehabilitation. Hyperventilation and hypoventilation, inhibited breathing, and breath suspension are all deviations from an optimal breathing pattern in which breathing volume is closely matched to metabolic needs. Such disordered breathing has varying effects on acid/base balance, arterial diameter, and sodium retention by the kidneys. Therefore, a chronic breathing imbalance can contribute to pathophysiology, which may be remediable to an extent by altering habitual breathing patterns. PMID- 14531163 TI - Behavioral interventions in asthma. Breathing training. AB - Breathing exercises are frequently recommended as an adjunctive treatment for asthma. A review of the current literature found little that is systematic documenting the benefits of these techniques in asthma patients. The physiological rationale of abdominal breathing in asthma is not clear, and adverse effects have been reported in chronic obstructive states. Theoretical analysis and empirical observations suggest positive effects of pursed-lip breathing and nasal breathing but clinical evidence is lacking. Modification of breathing patterns alone does not yield any significant benefit. There is limited evidence that inspiratory muscle training and hypoventilation training can help reduce medication consumption, in particular beta-adrenergic inhaler use. Breathing exercises do not seem to have any substantial effect on parameters of basal lung function. Additional research is needed on the psychological and physiological mechanisms of individual breathing techniques in asthma, differential effects in subgroups of asthma patients, and the generalization of training effects on daily life. PMID- 14531164 TI - Breathing training for treating panic disorder. Useful intervention or impediment? AB - Breathing training (BT) is commonly used for treatment of panic disorder. We identified nine studies that reported the outcome of BT. Overall, the published studies of BT are not sufficiently compelling to allow an unequivocal judgment of whether such techniques are beneficial. This article discusses problems with the underlying rationale, study design, and techniques used in BT, and it identifies factors that may have determined therapy outcomes. The idea that hypocapnia and respiratory irregularities are underlying factors in the development of panic implies that these factors should be monitored physiologically throughout therapy. Techniques taught in BT must take account of respiration rate and tidal volume in the regulation of blood gases (pCO2). More studies are needed that are designed to measure the efficacy of BT using an adequate rationale and methodology. Claims that BT should be rejected in favor of cognitive or other forms of intervention are premature. PMID- 14531165 TI - Liver support in acute liver failure. PMID- 14531166 TI - To work in a hospital--a new risk factor for development of venous disease? PMID- 14531167 TI - [The double helix is 50]. PMID- 14531168 TI - [Malignant hyperthermia]. AB - Malignant hyperthermia is an autosomal dominant disorder of the skeletal muscle that predisposes affected individuals to a life-threatening hypermetabolic reaction in response to volatile anaesthetics and depolarizing muscle relaxants. The underlying heterogeneous genetic defects are mainly point mutations within the ryanodine receptor gene of the sarcoplasmic reticulum. Following the introduction of efficient diagnostic and therapeutic tools--the in vitro contracture test and intravenous treatment with dantrolene--a dramatic decline in mortality rates has been observed. The association of malignant hyperthermia-like reactions with other neuromuscular disorders requires the collaboration of several clinical disciplines to achieve a timely recognition of this still life threatening disorder. PMID- 14531170 TI - High prevalence of chronic venous disease in hospital employees. AB - OBJECTIVES: Chronic venous disease (CVD), which comprises primary/idiopathic abnormalities of the venous system, and secondary sequels after deep venous thrombosis are major health issues in Western countries. The present study was conducted to prove the hypothesis that the development of CVD might be triggered by exogenous, occupational risk factors. METHODS: We determined the prevalence and social relationship of CVD in a wide cross-section of hospital employees (n = 209; medical doctors and nurses, medical technician assistants, secretaries, scientific staff, cleaners and utility workers) without predocumented CVD. Prevalence, known endogenous risk factors for CVD and occupational and environmental risk factors (family history of venous disease, history of deep venous thrombosis, current oral contraceptive therapy, obesity, regular participation in sports or frequent use of saunas or sun-baths, and long periods of standing during work) were investigated. The restriction in quality of life due to symptoms of CVD was also evaluated. CVD was classified according to the CEAP criteria. RESULTS: CVD was present in a total of 70 employees (34%), predominantly in women. Standing at work was a predisposing factor. We found the highest prevalence of CVD in utility workers and cleaners and the lowest prevalence in medical technician assistants, secretaries and scientific staff. CONCLUSION: The study demonstrated that within a representative cross-section of hospital employees in a University hospital the prevalence of CVD was highest in women, especially in those working in a standing position or under conditions of high temperature and humidity. The results warrant regular evaluation of risk factors with subsequent primary prophylaxis of CVD. PMID- 14531169 TI - Cellular and molecular themes in apoptosis. AB - Apoptosis, an active mechanism of cell death, is of central importance in many biological scenarios. Research in this area has the potential to contribute to our understanding of many diseases and raises several potential therapeutic opportunities. Given this potential and the speed with which our understanding of this field has advanced over recent years, it is timely to introduce the clinician to the background on which the clinical implications of this research will be built. This review begins with contrasting apoptosis with the other mechanism of cell death, necrosis, and then outlines the features by which apoptosis may be recognised. With a view to understanding the level at which this process may be involved in disease and therapeutics, it is important to be aware of the basic mechanistic features of the induction and execution of apoptosis. In this, surface molecules such as CD95 (Fas) and the cascade of intracellular enzymes involved at many levels in apoptosis, the caspases, are of central importance. In all this, the mitochondrion is crucial to the induction of apoptosis and the regulation of the whole process. In the last part of this review, we attempt to draw out the clinical relevance of all this information. It is clear that apoptosis has an important role in the pathophysiology of malignancy, particularly with respect to haematological cancers, but also other oncological diseases. Apoptosis is also very important in autoimmune disease and viral infection. Finally, it is clear that apoptosis may be manipulated therapeutically to the benefit of patients in various scenarios. This is clearly an exciting area for future development, but one which clearly depends on a thorough mechanistic understanding. PMID- 14531172 TI - [Ruptured abdominal aortic aneurysms: status quo after a quarter century of treatment experience]. AB - BACKGROUND: Postoperative mortality rates between 40% and 50% have been invariably reported for the treatment of ruptured abdominal aortic aneurysms (rAAA) over the last 50 years. The aim of this analysis was to investigate which patient subgroups benefit from open surgery and in which subgroups a change of treatment strategies should be considered due to lack of improvement despite optimal patient management. PATIENTS AND METHODS: From 1980 to 2002 a total of 230 patients underwent surgery because of a ruptured AAA. The observation period was divided into 3 intervals to achieve an approximately equal distribution of patients. The effect of the observation period and of baseline parameters on mortality rates were investigated. RESULTS: Between 1980 and 1990, 72 patients were operated with a mortality rate of 38.9% (n = 28). During the second period (1991-1996) surgery was performed in 72 patients with a mortality rate of 40.3% (n = 29). In the third observation interval (1997-2002) 86 patients underwent surgery with an unvaried high mortality rate of 40.7% (n = 35). By applying a logistic regression model including age, gender, modality of rupture, location of cross-clamping and type of operation, only the modality of rupture and the patient's age, which are uncontrollable by the surgeon, could be shown to have a significant impact. CONCLUSION: Summing up these findings, open surgical repair of rAAA only leads to acceptable results when performed in younger patients without supposed comorbidities. Survival appears to be accidental in patients with advanced age and increased prevalence of relevant comorbidities/underlying diseases. Minimally invasive techniques may offer promising treatment options to those patients, as they do in elective interventions. PMID- 14531171 TI - [Occult scaphoid-lunate dissociation in distal radius fractures]. AB - INTRODUCTION: The aim of this study was to investigate the clinical consequences of scapholunate dissociation associated with fractures of the distal radius and the impact on wrist function. Fractures of the distal radius and scapholunate dissociation overlap in pathomechanics. The diagnosis however is frequently missed initially. PATIENTS AND METHODS: We reviewed 45 consecutive patients with closed distal radius fractures with a mean follow up of 48 months. Ten patients underwent surgery and 35 patients were treated by fracture reduction and cast immobilization during 4.5 (range 3 to 8) weeks. All patients were re-examined clinically and radiographs of both wrists were compared. RESULTS: Four patients showed evidence of SLD in the scapholunate joint region based on specific criteria (scapholunate gaps > 2 mm on anterior-posterior radiograph and the scapholunate angle > 60 degrees on lateral radiograph). All patients with SLD showed a poor radioulnar deviation. Three patients reported mild to moderate pain. DISCUSSION: The difficulties in management of SLD may be avoided by early detection and treatment. In all investigated patients the diagnosis was missed after the initial trauma. Untreated SLD can lead to carpal collapse and arthrosis of the wrist, and ultimately to scapholunate advanced collapse. PMID- 14531173 TI - Efficacy and tolerability of non-invasive ventilation delivered via a newly developed helmet in immunosuppressed patients with acute respiratory failure. AB - OBJECTIVES: To assess efficacy and tolerability of a newly developed helmet for the delivery of non-invasive ventilation in patients with acute respiratory failure. PATIENTS AND METHODS: Ten consecutive immunocompromised patients with acute respiratory failure admitted to our intensive care unit were included in the study. The patients were equipped with the helmet and non-invasive ventilation (NIV) was performed. Oxygenation and tolerability were assessed during the first 24 hours of NIV. RESULTS: All patients tolerated the helmet well and their oxygenation improved. Two patients developed septic shock and had to be endotracheally intubated during the study period, eight patients survived to be weaned from NIV. CONCLUSIONS: NIV delivered via the helmet is effective and may serve as a better tolerated alternative to endotracheal intubation and to NIV via a standard face mask. PMID- 14531174 TI - Liver support in fulminant liver failure after hemorrhagic shock. AB - Acute liver failure (ALF) is a rare clinical syndrome associated with a mortality of up to 80% and its management remains an interdisciplinary challenge. Despite recent improvements in intensive care management, the mortality of patients with ALF remains high and is related to complications such as cerebral edema, sepsis and multiple organ failure. Emergency orthotopic liver transplantation (OLT) is currently the only effective treatment for those patients who are unlikely to recover spontaneously. Nevertheless, OLT is not always possible because of the shortage of the organs and/or complications related to ALF. Newly introduced liver-assist devices can temporarily support the patient's liver until native liver recovers or can serve as a bridging device until a liver graft is available. The support devices use both cell-based and non-cell-based techniques. One of the latest non-cell-based extracorporeal hepatic support devices, the molecular adsorbent recycling system (MARS), is based on the concept of albumin dialysis. MARS utilises selective hemodiafiltration with countercurrent albumin dialysis aiming to selectively remove both water-soluble and albumin-bound toxins of the low and middle molecular-weight range. We report on a young patient who presented with clinical symptoms of ischemic hepatitis and multi-organ failure (APACHE II score 38-->predicted postoperative mortality 87%) due to prolonged hemorrhagic shock. OLT was contraindicated because of history of pancreas cancer with metastases. It was necessary to use aggressive conservative therapy and an extracorporeal liver-assist device until liver regeneration began and hemodynamic conditions were stable. The patient underwent five treatments with MARS. During the treatment, there were improvements of hemodynamics, respiratory function, acid-base disturbances and laboratory parameters. The plasma disappearance rate of indocyanine green, a parameter of dynamic liver function, improved during MARS treatment. Although repeated neurological examination predicted diffuse brain damage (brain oedema, decreased cerebral blood flow), the patient recovered without any neurological deficits. The patient survived and was discharged from the hospital in good condition. In this case MARS treatment was successful in supporting the patient through the most critical period of ALF. PMID- 14531175 TI - Successful treatment of refractory cerebral oedema in ecstasy/cocaine-induced fulminant hepatic failure using a new high-efficacy liver detoxification device (FPSA-Prometheus). AB - Ecstasy-induced fulminant hepatic failure is associated with high mortality. If complicated by cerebral oedema, orthotopic liver transplantation is the only established treatment. We report a case of combined ecstasy/cocaine-induced fulminant hepatic failure presenting with severe rhabdomyolysis, myocardial infarction and multiorgan failure. Transplantation was declined by the transplant surgeons because of a history of intravenous drug abuse. As excessive hyperammonaemia (318 mumol/l) and refractory transtentorial herniation developed, treatment with a new liver detoxification device combining high-flux haemodialysis and adsorption (FPSA-Prometheus) was initiated. Within a few hours of treatment, ammonia levels normalised. Cerebral oedema was greatly reduced by day 4 and hepatic function gradually recovered. Following neurologic rehabilitation for ischaemic sequelae of herniation, the patient was discharged from hospital with only minimal deficits. In conclusion, efficient extracorporeal detoxification may be an option for reversal of hyperammonaemia and refractory cerebral oedema in ecstasy/cocaine-induced acute liver failure. PMID- 14531177 TI - Is there enough rationale in randomised controlled trials on distant healing? PMID- 14531176 TI - [Consensus recommendations on drug treatment of bronchial asthma in children and adolescents. 1. Addendum (2003). Austrian Society for Pediatrics and Adolescent Medicine and Austrian Society for Lung Diseases and Tuberculosis]. PMID- 14531179 TI - Small community hospital turnaround requires niche marketing. PMID- 14531178 TI - Seeking a return to profitability. AB - This special report by turnaround expert Dr. Brian Goodell examines the financial signs that spark the need for a turnaround. Learn how a turnaround strategy works and, perhaps most importantly, how a health care organization can survive a turnaround. PMID- 14531180 TI - No lollygagging for Dr. Lally. AB - Jim Lally considers himself something of a late-bloomer because he spent years in the Army before earning his bachelor's degree and going on to medical school. But he's making up for lost time quickly. He already owns his own hospital. PMID- 14531181 TI - Zen and the art of dealing with difficult physicians. A three-fold path for enlightened leaders. PMID- 14531182 TI - Naked reverse: unexpected plays for exceptional results. AB - Following the herd is easy when it comes to business ventures. But if you really want to start something new, you must first carefully think it through. Learn some ways to do that before you set out on a new path. PMID- 14531184 TI - A business case for patient safety. PMID- 14531183 TI - The art and science of winning physician support for Six Sigma change. AB - Active physician participation and acceptance of Six Sigma is critical to successful implementation. Take a look at strategies and techniques that can help secure physician support for Six Sigma. PMID- 14531186 TI - Are you on the right track? AB - Physicians in early or mid-level leadership positions need different skills than those at the top. What are the skills they need and what are the risks of focusing on the wrong skills? PMID- 14531187 TI - The legacy of Ladan and Laleh. PMID- 14531185 TI - Turning data into information: the cost of making diamonds from coal. PMID- 14531188 TI - 10 ways to ensure optimal management of your practice. PMID- 14531189 TI - 10 factors affecting the physician shortage of the future. PMID- 14531190 TI - Make forward progress: master lateral management. PMID- 14531191 TI - Using the balanced scorecard as a performance management tool. PMID- 14531192 TI - The battle for a prescription drug benefit. PMID- 14531193 TI - [The Medical Association of Puerto Rico]. PMID- 14531195 TI - Surgical management of morbid obesity: the Puerto Rico experience. PMID- 14531194 TI - [Cancer of the superior pulmonary sulcus]. PMID- 14531196 TI - Necrotizing enterocolitis. PMID- 14531197 TI - Sentinel node biopsy in invasive breast cancer and ductal carcinoma in situ. PMID- 14531198 TI - [Graduate medical education at the University of Puerto Rico Medical Sciences Campus confronting health care reform]. PMID- 14531199 TI - [The history of surgery in Puerto Rico]. PMID- 14531200 TI - Facility profile. A calming place for cancer outpatients. PMID- 14531201 TI - A higher goal. Evidence-based design raises the bar for new construction. PMID- 14531202 TI - Rapid response. A Toronto-area health facility manager recounts his experiences with the SARS scare. PMID- 14531203 TI - IT update. Telemedicine applications grow, while data security remains a top priority. PMID- 14531204 TI - Pure + easy. Selecting a domestic water disinfection system. PMID- 14531205 TI - Sneak peek. A look at JCAHO's reorganized Environment of Care chapter. PMID- 14531206 TI - Latest in lavs. Rest room designs are changing, and cleaning methods are following suit. PMID- 14531207 TI - Perspectives. What's driving spending? More stuff, no brakes. PMID- 14531208 TI - The management of septic arthritis. AB - Septic arthritis--infection with resulting inflammation within a joint--is an emergency. Not only can it rapidly destroy a joint or irreversibly impair joint function, it may be fatal. So although the condition is not common, affecting between 2 and 10 per 100,000 people each year in the UK, it must be recognised and tackled promptly. Here we review the management of septic arthritis, particularly that of bacterial origin. PMID- 14531209 TI - Is Cerazette the minipill of choice? AB - Around 5% of women aged 16-49 years in Great Britain use a progestogen-only pill (POP; 'minipill') as contraception. These pills are used as alternatives to combined oral contraceptives (COCs), compared to which they are less reliable at preventing pregnancy: the estimated contraceptive failure rate of POPs is 0.5 pregnancies per 100 woman-years when used consistently and correctly, compared with 0.1 per 100 woman-years for COCs. Cerazette (Organon), a new POP, is being promoted by the company as "the first oestrogen free pill to consistently inhibit ovulation", as having "the efficacy of a combined pill, with the reassurance of an oestrogen free pill" and offering "reliable contraception for women of any reproductive age". Here, we consider whether Cerazette offers advantages over established POPs. PMID- 14531210 TI - Bosentan for pulmonary arterial hypertension. AB - Pulmonary arterial hypertension is an uncommon but disabling and often fatal condition, in which there is a sustained rise in pulmonary arterial pressure due to progressive obliteration of the pulmonary vascular bed. [symbol: see text] Bosentan (Tracleer-Actelion), which belongs to a new class of drugs called endothelin receptor antagonists, is now available for treating patients with pulmonary arterial hypertension. Here we assess whether bosentan offers worthwhile benefits in the management of patients with this condition. PMID- 14531211 TI - Arthur Guyton, sodium, and hypertension. PMID- 14531212 TI - Neuroendocrine responses to stressors in lactating and nonlactating mammals: a literature review. AB - Although stress research is a popular topic of study, little is known about the neuroendocrine responses to a stressor in lactating and nonlactating humans. The purpose of this systematic review of the literature was to examine the neuroendocrine responses, specifically the glucocorticoids and catecholamines, in lactating and nonlactating animals and humans to an acute stressor. A brief overview of the physiological stress response in the human is included. Animal studies strongly suggest that lactation is associated with major changes in neuroendocrine responses to a variety of acute stressors. Neuroendocrine responses in humans to stressors are less clear due to the limited research. Future research is needed involving these responses in humans generally, as well as specifically in the patterns of neuroendocrine responses to chronic stressors in lactating and nonlactating women. PMID- 14531213 TI - Insulin resistance and obstructive sleep apnea: is increased sympathetic stimulation the link? AB - The science of sleep is in early stages of development, and the biochemical consequences of obstructive sleep apnea (OSA) are slowly being identified. Only recently have investigators begun to identify the commonalities and interaction between OSA and insulin resistance, the underlying pathology of type 2 diabetes. Obesity and increasing age play important parts in the natural history of both conditions, which frequently coexist. The purpose of this article is, first, to examine the extent and strength of studies that have investigated the association between OSA and increased insulin resistance or type 2 diabetes and, second, to propose a model that explains the association and cyclical interaction between OSA, obesity, and insulin resistance. PMID- 14531214 TI - A candidate locus approach identifies a long QT syndrome gene mutation. AB - Long QT syndrome is an inherited disorder that results in lengthened cardiac repolarization. It can lead to sudden onset of torsades de pointes, ventricular fibrillation, and death. The authors obtained a family history, performed electrocardiograms, and drew blood for DNA extraction and genotyping from 15 family members representing 4 generations of an affected family. Seven individuals demonstrated prolonged QT intervals. The authors used polymorphic short tandem repeat markers at known LQTS loci, which indicated linkage to chromosome 11p15.5 where the potassium channel, KCNQ1, is encoded. Polymerase chain reaction was used to amplify the coding region of KCNQ1. During survey of the KCNQ1 coding region, a G-to-A transition (G502A) was identified. DNA from all clinically affected but from none of the clinically unaffected family members carried the G-to-A transition. The candidate locus approach allowed an efficient mechanism to uncover the potassium channel mutation causing LQTS in this family. PMID- 14531215 TI - Effects of morphine and time of day on pain and beta-endorphin. AB - Clients report more pain at some times of day than at others due, in part, to the temporal variation of the body's inhibitory pain response. The analgesic effectiveness of morphine varies with the time of day, perhaps due to the inhibiting or enhancing effects of the drug on plasma beta-endorphin (BE). This experiment was designed to examine the timed effects of morphine on the pain induced BE response. Six groups of treatment mice (injected with morphine sulfate) and 6 groups of control mice (injected with saline) were exposed to an acute pain stimulus at 4-h intervals, and blood was collected. Plasma BE was analyzed using radioimmunoassay. Control mice showed a robust circadian BE response rhythm with a peak at 0000 and a nadir at 1200, whereas the BE response of mice that received morphine was arrhythmic. Animals that received morphine tolerated the noxious stimulus longer, but the analgesia varied with time of day. These results indicate that morphine abolishes the rhythmic BE response to pain and does not inhibit pain equally at all times of day. Morphine doses should be titrated to maximize the endogenous pain control system while achieving analgesia with decreased dosages. PMID- 14531216 TI - Endocytosis and signal transduction: basic science update. AB - Endocytosis can be separated into the categories of phagocytosis and pinocytosis. Phagocytosis can be distinguished from pinocytosis primarily by the size of particle ingested and by its dependence on actin polymerization as a key step in particle ingestion. Several specific forms of pinocytosis have been identified that can be distinguished based on their dependence on clathrin or caveolin. Both clathrin- and caveolin-dependent pinocytosis appear to require the participation of dynamin to internalize the plasma membrane. Other, less well-characterized forms of pinocytosis have also been described. Although endocytosis has long been known to affect receptor density, recent studies have demonstrated that endocytosis through clathrin- and caveolin-dependent processes plays a key role in receptor-mediated signal transduction. In some cases, blockade of these processes attenuates, or even prevents, signal transduction from taking place. This information, coupled with a better understanding of endocytosis mechanisms, will help advance the field of cell biology as well as present new targets for drug development and disease treatment. PMID- 14531217 TI - Association of proinflammatory molecules with apoptotic markers and survival in critically ill multiple organ dysfunction patients. AB - Recent evidence supports the involvement of apoptosis in multiple organ dysfunction (MODS). The authors examined the hypothesis that nitric oxide (NO), interleukin (IL)-6, tumor necrosis factor (TNF)-alpha, and cortisol correlate with Fas and Fas ligand (FasL) expression on peripheral blood mononuclear cells and that Fas and FasL, therefore, mediate their association with MODS severity. Thirty-five critically ill adult MODS patients were followed for up to 14 days and were compared to non-MODS matched controls. Fas, FasL, nitrate, cortisol, and IL-6 were elevated in MODS patients (P < 0.05). Nitrate and cortisol correlated with Fas expression (P < 0.05). All factors studied, except for TNF-alpha, correlated with MODS severity (P < 0.05); however, by multivariate analyses, Fas and FasL were independently associated with severity and survival (P < 0.05). The inflammatory molecules studied may mediate the association of apoptotic constituents with MODS severity and survival only in part. PMID- 14531219 TI - Is your document control out of control? Complying with document control regulations. AB - Clinical laboratories, faced with new or changing regulations and shortages in trained laboratory staff, may find it difficult to comply with guidelines related to document control, upkeep of standard operating procedure manuals, and documentation of quality programs or competency reviews. Now, more than ever, it is critical to keep documents, procedures, and training material organized and accessible to busy laboratory staff and inspectors. The key may be the ability to use existing software programs available to industry for similar compliance issues. The laboratory's ability to change and update quickly with limited staffing may depend on electronic solutions to the complexities of document control. Several software programs have merit and can be integrated into a clinical laboratory. PMID- 14531218 TI - Natural killer cell cytotoxicity: a methods analysis of 51chromium release versus flow cytometry. AB - The purpose of this study is to describe design considerations for the use of flow cytometry (FC) compared to 51chromium (51Cr)-release assays utilizing cryopreserved peripheral blood mononuclear cells (PBMCs) to detect natural killer (NK) cell cytotoxicity. Subjects were 10 healthy women aged 18 to 39 years. Intra assay variability between methods differed only at the lowest effector-target ratios evaluated. Interassay variability was wide but did not differ between methods. The relationship of lytic unit-10 between methods was strongly positive. Cytotoxicity detected by 51Cr release was higher than that detected by FC for all 10 subjects. Cost was comparable. However, had more assays been performed, technician time would have been greater with flow cytometry. More whole blood was needed to perform the flow cytometry cytotoxicity assay than 51Cr-release cytotoxicity assay. The authors found no compelling reason to adopt NK cell cytotoxicity by flow cytometry over 51Cr release. PMID- 14531220 TI - Evaluation of rule-based autoverification protocols. AB - Autoverification is a technique that can help save time and dollars in today's modern laboratory. Most laboratorians, however, are afraid to use it. The authors describe a rules-based system based on the quality of the laboratory's instrumentation and the quality of the results generated to help decide if the laboratory should pursue the advantages of autoverification. This project was developed through a grant from Ortho-Clinical Diagnostics, Inc., a Johnson & Johnson Company. PMID- 14531221 TI - Promoting success: educational interventions for students with special needs. PMID- 14531222 TI - Detection of the sentinel anthrax case in the United States. AB - First-hand knowledge of the detection of the first bioweapon in modern United States history is described in this article. The method by which the presumptive diagnosis of anthrax meningitis was made within 13 hours of the patient presenting to the emergency department is described using pre-analytic, analytic, and post-analytic phases. The lessons learned from this process are briefly presented so that other laboratories may learn from our experience: how to prepare; how to quickly analyze a potential bioweapon; how to communicate with staff and local, regional, and national authorities; and how to deal with disruptive media attention. PMID- 14531223 TI - Career development strategies. The stages of a manager's life, Part II. PMID- 14531224 TI - Managing personnel: the importance of procedural fairness. PMID- 14531226 TI - Designing for robotics. PMID- 14531225 TI - Common cent$ 14: Pleasure, pain, and the economics of motivation. PMID- 14531227 TI - Where are we going? Health-care professionals and the staffing crisis. PMID- 14531228 TI - Benchmarking for excellence. PMID- 14531229 TI - Secrets to hiring the right person for your laboratory. PMID- 14531231 TI - National briefing: science and health--influenza vaccine supply expected to meet demand, CDC recommends influenza vaccinations begin in October. PMID- 14531230 TI - The sural sensory nerve is usually spared in Guillain-Barre syndrome. AB - Sensory nerve conduction (NC) studies that were performed on patients with Guillain-Barre syndrome (GBS) were reviewed. These were compared with the sensory NC findings in patients with peripheral neuropathy of the axonal-loss type (axonopathies) and chronic inflammatory demyelinating polyneuropathy (CIDP). In GBS, the sural sensory nerve frequently showed normal NC findings compared to the median and ulnar sensory nerves. The reverse pattern was seen in axonopathies and CIDP, in which the sural nerve showed more abnormalities than the median and ulnar nerves. This pattern of sensory NC abnormalities may assist the examiner in differentiating GBS from other types of peripheral polyneuropathy. PMID- 14531232 TI - Looking back: the surgical treatment of idiopathic epilepsy. May 8-9, 1917. PMID- 14531233 TI - One-year hospital pharmacy residencies retain eligibility for special Medicare payment. PMID- 14531234 TI - Virginia HRSA project underway. PMID- 14531235 TI - Michigan health system focuses on medication safety. PMID- 14531236 TI - Medco becomes standalone firm. PMID- 14531237 TI - New drugs and dosage forms. PMID- 14531239 TI - Pharmacists' role in preventing and treating substance abuse: why are we doing so little? PMID- 14531238 TI - Use of a personal digital assistant in a pharmacy-directed warfarin dosing program. PMID- 14531240 TI - Current and emerging therapies for the management of chronic inflammation in asthma. AB - The pathophysiology of asthma, current treatment guidelines, and emerging immunomodulating treatments are reviewed. Asthma is a chronic disease of inflammation, which left untreated, may result in irreversible lung damage. Inhaled antigens elicit a T-helper cell type-2 response, leading to the production of immunoglobulin E (IgE) and eosinophil sensitization. Eosinophil infiltration into the airway is thought to play an important role in chronic inflammation and increases several weeks before an acute exacerbation in moderate to severe asthma. The updated guidelines from the National Asthma Education and Prevention Program and the National Institutes of Health focus on the use of daily antiinflammatory treatment in patients with persistent asthma. Inhaled corticosteroid (ICS) therapy remains the cornerstone for controlling mild and moderate asthma, but when used in higher doses for moderate-to-severe asthma it may cause long-term adverse effects, including osteoporosis and glaucoma. Step-up therapy should be the combination of a long-acting beta-agonist (LABA) and an ICS. Leukotriene modifiers may provide additional symptom control in patients with moderate to severe asthma but have not been shown to be as cost-effective as monotherapy. Newer entities focus on direct immune modulation and include monoclonal antibodies targeting various inflammatory-response receptors (IgE, interleukin [IL]-2, IL-4, and IL-5). Phosphodiesterase-4 (PDE-4) inhibitors may be effective for chronic obstructive pulmonary disease, but have not delivered consistent results for controlling asthma. Cytokine-receptor antagonists and leukocyte-suppressing antiinflammatory drugs (LSAIDs) are currently under investigation and have yielded promising preliminary results as alternatives for the treatment of asthma. Current and emerging therapies for the management of chronic inflammation in asthma include ICSs, LABAs, leukotriene modifiers, monoclonal antibodies, PDE-4 inhibitors, cytokine-receptor antagonists, and LSAIDs. PMID- 14531241 TI - Effect of fluoroquinolone expenditures on susceptibility of Pseudomonas aeruginosa to ciprofloxacin in U.S. hospitals. AB - The effect of fluoroquinolone use on the susceptibility of Pseudomonas aeruginosa to fluoroquinolones in U.S. hospitals was studied. Benchmarking surveys were sent annually to pharmacists practicing in U.S. hospitals from 1993 to 1999. Data collected included hospital characteristics, antimicrobial expenditures and use, antimicrobial stewardship activities, and bacterial susceptibilities. Antimicrobial expenditures were normalized for the number of occupied beds (OBs) per year. General linear modeling and repeated-measures mixed-effects modeling were used to determine factors predictive of P. aeruginosa susceptibility to fluoroquinolones. A total of 174 hospitals provided data for fluoroquinolone expenditures and susceptibility of P. aeruginosa; the median number of years of data was 3 (range, 1-6), representing 416 hospital years. Community hospitals contributed a majority of the data. Median fluoroquinolone expenditures increased gradually from $230 per OB in 1993 to $400 per OB in 1998. A 55% increase to $620 per OB occurred in 1999, largely because of increased spending on levofloxacin. Susceptibility to ciprofloxacin was commonly used to assess fluoroquinolone susceptibility. The median susceptibility of P. aeruginosa to ciprofloxacin decreased from 84% to 71%. Increasing expenditures for ofloxacin and levofloxacin, but not ciprofloxacin, were associated with decreasing P. aeruginosa susceptibility to ciprofloxacin. In the final multivariable model, each study year after 1993 and every increase in ofloxacin expenditure of $100 per OB were associated with decreases in P. aeruginosa susceptibility. Data from a benchmarking survey of U.S. hospitals for 1993-1999 revealed increases in levofloxacin expenditures, total fluoroquinolone expenditures, expenditures for nonfluoroquinolone antipseudomonal antimicrobials, and total antimicrobial expenditures in 1999. Increases in expenditures for levofloxacin and ofloxacin were associated with a significant decrease in P. aeruginosa susceptibility to ciprofloxacin. PMID- 14531242 TI - Changes in heart rate associated with nebulized racemic albuterol and levalbuterol in intensive care patients. AB - The effects of equipotent doses of racemic albuterol and levalbuterol on heart rate (HR) in intensive care patients with and without baseline tachycardia were studied. Patients were included if they were hemodynamically stable and required bronchodilator therapy every four hours; patients were excluded if they were maintained on a beta-blocker. Four hours after the most recent bronchodilator treatment, each patient was randomized to receive at least two consecutive doses of albuterol 2.5 mg or levalbuterol 1.25 mg four hours apart via nebulization. HR was recorded at the end of the second dose and 5, 10, 15, 30, 60, 90, 120, 180, and 240 minutes after treatment. Twenty intensive care patients, including 10 with baseline tachycardia and 10 without baseline tachycardia, were enrolled. In patients with baseline tachycardia, the mean largest HR increase was 1.4 beats/min (1.3%) with albuterol and 2.0 beats/min (2.1%) with levalbuterol (both increases were not significant). In patients without baseline tachycardia, the mean largest HR increase was 4.4 beats/min (6.7%) with albuterol (p = 0.04) and 3.6 beats/min (5.0%) with levalbuterol (p = 0.03). Short-term use of nebulized albuterol and levalbuterol was associated with similar changes in HR in intensive care patients with or without baseline tachycardia. PMID- 14531243 TI - An Internet-based store-and-forward video home telehealth system for improving asthma outcomes in children. AB - The adherence and disease-control outcomes associated with the use of an Internet based store-and-forward video home telehealth system to manage asthma in children were studied. Pediatric patients with persistent asthma were provided with home computers and Internet access and monitored biweekly over the Internet. All patients were seen in the pediatric clinic at 0, 2, 6, 12, and 24 weeks. Half of the patients received asthma education in person and half via an interactive Web site. Adherence measures were assessed by therapeutic and diagnostic monitoring. Therapeutic monitoring included digital videos of patients using their controller medication inhaler. Diagnostic monitoring included an asthma symptom diary and a video of peak flow meter use. Videos were submitted electronically twice a week by using in-home telemonitoring with store-and-forward technology. Feedback was provided electronically to each patient. Disease control was assessed by examining quality of life, utilization of services, rescue-therapy use, symptom control, satisfaction with home telemonitoring, and retention of asthma knowledge. Patients were randomly assigned to an asthma education group (Internet versus office), and the data were analyzed by comparing results for study days 0 90 and 91-180. Ten children participated. A total of 321 videos of inhaler use and 309 videos of peak flow meter use were submitted. Inhaler technique scores improved significantly in the second study period. Submission of diagnostic monitoring videos and asthma diary entries decreased significantly. Peak flow values as a percentage of personal best values increased significantly. Overall, there was no change in quality of life reported by patients. However, the caregivers in the virtual-education group reported an increase in the patients' quality-of-life survey scores. Emergency department visits and hospital admissions for asthma were avoided. Rescue therapy was infrequent. A high rate of satisfaction with home telemonitoring was reported. Internet-based, store-and forward video assessment of children's use of asthma medications and monitoring tools in their homes appeared effective and well accepted. PMID- 14531244 TI - Program using pharmacy technicians to obtain medication histories. PMID- 14531245 TI - Initiative for improving pharmacist satisfaction with work schedules. PMID- 14531246 TI - ASHP statement on the role of health-system pharmacists in emergency preparedness. PMID- 14531248 TI - ASHP statement on the pharmacist's role in the care of patients with HIV infection. PMID- 14531247 TI - ASHP statement on the pharmacist's role in substance abuse prevention, education, and assistance. PMID- 14531249 TI - Professional casualties in America's war on drugs. PMID- 14531250 TI - Evaluation of propofol infusion syndrome in pediatric intensive care. PMID- 14531251 TI - [Optimal application time of lidocaine adhesive tape examined using current perception threshold measurement]. AB - BACKGROUND: We often use lidocaine adhesive tape (Penles, Wyeth Lederle Japan, Ltd., Tokyo, Japan) as a topical anesthetic prior to puncturing a vein with a needle. Since the tape is usually in place for a long time, we often experience problems with creasing and flaring when inserting the needle. We attempted to determine the optimal time for application of the tape as well as for vein puncture after its removal by measuring pain sensation in human subjects. METHODS: Lidocaine tape was applied to the dorsum of volunteer hands for 2, 4, 6, and 12 hours, and we evaluated changes after removal by determining current perception threshold (CPT), pain score, touch sensation score, diameter of the vein, pruritus, and flare. RESULTS: CPT levels were found to increase within 1 hour after removal of the lidocaine tape with all 3 electrical stimulus rates used (2000 Hz, 250 Hz, 5 Hz). Further, pain and touch sensation scores were low within 2 hours of removal regardless of application time. Notably, with 4 and 6 hours of application, scores were stable 4 and 6 hours after removal. However, vein diameter and pruritus results did not change regardless of application time, while crease and flare were only observed within 30 minutes after removal. CONCLUSIONS: We concluded that lidocaine tape provided maximum pain relief when applied 4 to 6 hours prior to vein puncture and up to 1 hour after removal. PMID- 14531252 TI - [Spinal anesthesia using hyperbaric bupivacaine HCl for cesarean section]. AB - BACKGROUND: We performed this prospective study to determine the proper amount of hyperbaric bupivacaine hydrochloride as a spinal anesthetic agent for cesarean section. METHODS: The parturients were randomly allocated to receive one of four spinal agents in a blind manner; tetracaine 10 mg (control), bupivacaine 10, 12.5 and 15 mg. Morphine HCl 0.1 mg was added to each agent and the total volume was adjusted to 3.1 ml with 10% glucose solution. RESULTS: All the four spinal agents provided an adequate analgesic level (T 5) without serious complications. Among the three dosages of bupivacaine, the time interval requiring for anesthetic level to reach T 5 tended to be shorter with a larger amount of bupivacaine. The incidence of intraoperative supplemental analgesic and hypotension and the dosage of ephedrine used to treat hypotension were greater in the patients anesthetized with tetracaine 10 mg than in those anesthetized with bupivacaine 10 mg, which is equipotent to tetracaine 10 mg. CONCLUSIONS: 1. As a spinal anesthetic agent for cesarean section, hyperbaric bupivacaine is superior to tetracaine. 2. Hyperbaric bupivacaine 10 mg, 12.5 mg or 15 mg can be used safely and effectively as a spinal agent for cesarean section. 3. High dose bupivacaine is recommended in an urgent case, and low dose bupivacaine is recommended when maternal hypotension must be strictly avoided. PMID- 14531253 TI - [Prognostic factors in patients with massive intraoperative blood loss]. AB - BACKGROUND: Massive blood loss during surgery might affect the prognosis of the patient. METHODS: Fifty-seven patients with massive blood loss of more than 10,000 g were the subjects of the study. We compared the postoperative complication and prognosis for the following 3 groups. Group 1 is without complications (n = 17), Group 2 is alive with complications (n = 20), and Group 3 is the death group (n = 20). The following results were obtained. RESULTS: Transfusion, blood loss and urine output were prognosis decision factors. And we found out that the duration of low hemoglobin level (Hb < 7.0 g.dl-1) was closely related to the prognosis. CONCLUSIONS: It will be concluded that the duration of low hemoglobin level was one of the most important parameters to determine the prognosis of patients with massive intraoperative bleeding. PMID- 14531254 TI - [The relationship between postoperative body temperature for 24 hours and central nervous system dysfunction in patients of selective cerebral perfusion]. AB - BACKGROUND: Postischemic hyperthermia is known to exert detrimental effect on neurological outcome even 24 hours after brain ischemia in animal experiments. The purpose of this study is to investigate the effect of postoperative hyperthermia in cardiac surgery. METHODS: We studied consecutive patients in elective or emergency cardiac surgery with selective cerebral perfusion. We recorded body temperature (BT) during 24 postoperative hours and JCS at discharge from ICU. RESULTS: BT was higher in JCS III group than in JCS I and II group (P < 0.05). CONCLUSIONS: The results indicated that postoperative BT was associated with consciousness disturbance in cardiac surgery with selective cerebral perfusion. Significant BT elevation during postoperative 24 hours in ICU may be due to brain injury such as infarction or bleeding during selective cerebral perfusion. Postoperative hyperthermia may accelerate penumbra around infarcted area to necrosis, resulting in deep coma, which may imply a possibility of managing BT for further consciousness recovery. PMID- 14531255 TI - [Effect of body fat percentage on estimated propofol concentrations at awakening from anesthesia using target controlled infusion]. AB - BACKGROUND: Target controlled infusion (TCI) for propofol allows anesthesiologists to target constant blood concentrations of propofol. However, the pharmacokinetic parameters in TCI system do not take account of the patient's age, make up and gender. We evaluated the relationship between body fat percentage and the estimated effect site propofol concentrations at awakening. METHODS: Anesthesia was induced in 37 patients with fentanyl and propofol by TCI. Patients's percentage of fat was evaluated by measuring the thickness of the three parts of the skin fat with Skyndex (Caldwell Justiss, USA). After surgery and discontinuation of anesthesia, the estimated effect site propofol concentrations (EPEC) were determined when the patient could respond to verbal command. RESULTS: The EPECs at awakening were 1.5 +/- 0.2 micrograms.ml-1 in male and 1.5 +/- 0.3 micrograms.ml-1 in female. In male, the percentage of fat and body mass index (BMI) correlated significantly with EPEC (r = 0.79 and 0.49 respectively). In female, the percentage of fat and BMI did not correlate with EPEC. The estimated fentanyl effect site concentrations at awakening did not correlate with the EPEC. CONCLUSIONS: Male patients who have high percentage of fat have a tendency to delayed of awakening from propofol anesthesia. In female, however, no correlation was seen between percentage of fat and awakening time from propofol anesthesia. Measuring the percentage of fat might bring a new insight into pharmacokinetics of propofol. PMID- 14531256 TI - [Postoperative delirium after general anesthesia vs. spinal anesthesia in geriatric patients]. AB - BACKGROUND: Influence of the type of anesthesia on postoperative delirium was examined in geriatric patients with femoral neck fracture. METHODS: Forty patients aged 70 or more were randomly allocated to receive either general anesthesia (sevoflurane, nitrous oxide in oxygen, G group, n = 21) or spinal anesthesia (0.5% bupivacaine, S group, n = 19). G group received the oxygen therapy during the 12-hour postoperative period. Postoperative assessments included the rate of postoperative delirium during the 4-day postoperative period, oxygen saturation during the 18-hour postoperative period, the value of hemoglobin at 1 postoperative day and the number of analgesics requirements during the 4-day postoperative period. RESULTS: The rate of postoperative delirium was similar between the two groups during the 4-day postoperative period. However, that of S group tended to be higher during the 1-day postoperative period. Postoperative analgesic effect and the value of hemoglobin at 1 postoperative day were similar between the two groups. Oxygen saturation was similar between the two groups except 6 hours after the operation. CONCLUSIONS: The type of anesthesia, general or spinal, does not affect the postoperative delirium in geriatric patients with femoral neck fracture. PMID- 14531257 TI - [Sedation with intravenous midazolam during upper gastrointestinal endoscopy- changes in hemodynamics, oxygen saturation and memory]. AB - BACKGROUND: Cardiorespiratory adverse effects are often observed in patients undergoing upper gastrointestinal endoscopy with sedation. In this study, we examined hemodynamics, oxygen saturation and memory during upper gastrointestinal endoscopy under sedation with intravenous midazolam. METHODS: Eight healthy outpatients without any obvious complications received intravenous midazolam 5 mg for sedation for upper gastrointestinal endoscopy. Blood pressure, heart rate and percutaneous arterial oxygen saturation (SpO2) were measured before, during and after endoscopy. After the arousal by intravenous flumazenil, we inquired the patients about the level of memory during the endoscopy. RESULTS: Blood pressure decreased significantly two minutes after midazolam administration, but increased significantly after the insertion of an endoscope which was not different from the control value. Heart rate increased significantly one and three minutes after the insertion of the endoscope. SpO2 decreased significantly after midazolam administration and stayed at around 95%. No patients remembered the procedure. CONCLUSIONS: Sedation with intravenous midazolam during upper gastrointestinal endoscopy is useful to control the cardiovascular responses, and to obtain amnesia. However, a decrease in SpO2 should be watched carefully. PMID- 14531258 TI - [Anesthesia for a woman with triplet pregnancy presenting with acute abdomen after the recent epidural blood patch]. AB - We report a case of primipara with triplet pregnancy who underwent combined spinal and epidural anesthesia 10 weeks after epidural blood patch. At 15 weeks of gestation, a woman with triplet gestation underwent Shirodkar operation under spinal anesthesia and subsequent epidural blood patch as a treatment of post dural puncture headache. At 26 weeks she presented with acute abdomen and laparotomy was scheduled. Spinal anesthesia was selected with an epidural catheter inserted in case of prolonged operation and for postoperative pain control. The placement of an epidural catheter was without problem. Laparotomy revealed right paraovarian cyst torsion and the right salpingo paraoophocystectomy was performed. Patient-controlled analgesia with epidural bupivacaine and fentanyl was effectively continued for two days. Postoperative course was uneventful and the triplets were delivered by cesarean section at 35 weeks. PMID- 14531259 TI - [Sevoflurane anesthesia for pancreatectomy in an infant with nesidioblastosis]. AB - We report a case of nesidioblastosis, which is characterized by hyperinsulinism due to a diffuse increase in the number of beta-cells in the pancreas and, consequently, severe hypoglycemia. The patient was a 79-day-old boy. He had been suffering from severe hypoglycemia despite aggressive treatment, including glucose loading and administration of glucocorticoid and diazoxide. Pancreatectomy was performed. Anesthesia was induced by thiamilal 25 mg and vecronium and was maintained by 1 to 2% sevoflurane in 65% nitrous oxide and fentanyl 5 micrograms. No hemodynamic instability was observed during anesthesia. Blood glucose level was maintained around 200 mg.dl-1 without any hyperglycemic event by continuous infusion of 20% glucose at the rate of 14.5 mg.kg-1.min-1, which was calculated from the daily glucose demand to prevent hypoglycemia preoperatively. PMID- 14531260 TI - [Insertion of an epidural catheter from the surgical wound for postoperative analgesia--a case report]. AB - A 62-year-old male was scheduled for discectomy for lumbar disc herniation. He had bronchial asthma, and his asthma was induced by nonsteroidal anti inflammatory drugs. We decided to use continuous epidural block for postoperative analgesia. However, he was frightened of insertion of an epidural catheter. Therefore, we planned to insert the epidural catheter before the end of surgery under general anesthesia. It was easily placed at L 5-S 1 interspace using a Tuohy needle from the surgical wound, and the contrast medium 5 ml was injected through the epidural catheter to confirm the placement. After the surgery, continuous epidural block with local anesthetics was used for two days. He did not complain of severe pain postoperatively. PMID- 14531261 TI - [Effective radiofrequency lesioning for obturator nerve neuropathy]. AB - A 73-year-old female complained of pain in the right lower-abdomen and medial aspect of the distal thigh after a gynecological operation. Computer tomography and MRI findings excluded obturator canal herniation or lumbar plexopathy. Both lumbar epidural block and obturator nerve block with local anesthetics were effective, except their limited duration of pain relief and muscle weakness for several hours after the blockade. Treatment with radio-frequency lesion of 50 degrees C for 60 seconds of the right obturator nerve relieved her pain for 9 months without muscle weakness. This case shows usefulness of radiofrequency lesion for obturator nerve pain. PMID- 14531262 TI - [Anesthetic management with a laryngeal mask airway for gastrectomy in a patient with myotonic dystrophy]. AB - We report a case of 50-year-old male with myotonic dystrophy who underwent distal gastrectomy. A laryngeal mask airway (LMA) was inserted easily without using a muscle relaxant after propofol injection. Anesthesia was maintained with continuous propofol infusion and epidural anesthesia. The airway management with LMA enabled us to evaluate patient's awareness and respiratory pattern appropriately at the end of anesthesia. No respiratory complications, such as respiratory depression or atelectasis, occurred after surgery. We consider that LMA is useful for anesthetic management in a patient with myotonic dystrophy. PMID- 14531263 TI - [Iliopsoas abscess accompanied by epidural abscess--a case report]. AB - A 55-year-old man was admitted to a hospital with pain of the low back as well as the left leg, and fever. He was suspected of suffering from the lumbar disc herniation because of the presence of Lasegue's sign on the first physical examination. Abdominal computed tomography, however, revealed the swelling of the left iliopsoas muscle. Iliopsoas abscess accompanied epidural abscess was confirmed by subsequent magnetic resonance imaging (MRI). Antibiotic therapy was started for the successive 8 days. The fever resolved, but the pain persisted. The abscess extending from the iliopsoas muscle to the epidural space was still seen on the MRI 20 days after the completion of the antibiotic therapy, and he still complained of the pain of his low back and left leg. Therefore, we conducted epidural puncture under fluoroscopic guidance. Approximately 3 ml of pus was aspirated from the epidural space. Then, his complains decreased remarkably. Iliopsoas abscess should be taken into account in case of a patient with pain on the low back and leg and also inflammatory signs such as fever and leucocytosis. PMID- 14531264 TI - [A case of severe bradycardia and AV block during administration of propofol]. AB - A 69-yr-old man underwent emergency laparotomy. He was in endotoxic shock. Preoperative evaluation including a full blood count, chest X-ray and ECG were normal. Body temperature was 37.4 degrees C. Preoperative arterial pressure was 140/80 mmHg and heart rate 65 bpm. Anesthesia was induced with ketamine 100 mg, propofol 20 mg, fentanyl 50 micrograms and vecuronium 4 mg and maintained with propofol 4 mg.kg-1.hr-1 and fentanyl. Soon after opening the abdominal peritoneum, severe bradycardia (< 20 bpm) occurred, but it was effectively treated by ephedrine 16 mg. After that, surgery was performed uneventfully. In the intensive care unit (ICU), the patient developed four episodes of severe atrioventricular (AV) block after stimulation of the trachea by suction drainage under sedation with propofol, although there was no AV block during sedation with ketamine and propofol. After stopping propofol, the AV block was no longer observed. He was discharged from the ICU on the 12th postoperative day. Postoperative Holter ECG and echocardiography showed no abnormalities. It is likely that stimulation of the trachea triggered vagovagal reflex and propofol prolonged AV conduction, causing the AV block. PMID- 14531265 TI - [Heparin resistance during cardiopulmonary bypass]. AB - A 74-year-old man was scheduled for coronary artery bypass graft surgery with cardiopulmonary bypass. After intravenous heparin (200 U.kg-1), the activated clotting time (ACT) increased from 124 to 436 sec. However, it decreased to 128 sec immediately after cardiopulmonary bypass. The bypass was discontinued because the addition of heparin (200 U.kg-1) proved heparin resistance. The coronary artery bypass procedure was completed uneventfully without cardiopulmonary bypass. Several recent articles have reported that heparin resistance was corrected with antithrombin III concentrates, fresh frozen plasma, or argatroban. In this case, these drugs could not be used because the mechanism of heparin resistance remains uncertain. Thus, the off-pump technique is useful for unknown heparin resistance. PMID- 14531266 TI - [Postoperative complications and unanticipated admission in ambulatory surgery]. AB - BACKGROUND: Day-case surgery is becoming more popular in Japan. The incidence of postoperative complications and unanticipated hospital admission was evaluated between June 1, 1998 and May 31, 2001. METHODS: Two hundred fifty seven cases of ambulatory surgery were retrospectively evaluated in terms of methods of anesthesia, anesthesia time, recovery time, postoperative complications and unanticipated hospital admission. RESULTS: General anesthesia was used in 44% of 257 ambulatory surgical cases. Mean anesthesia time was 81 minutes and mean recovery time was 227 minutes. The most frequent complication was postoperative pain, incidence of which was 53%. The incidence of unanticipated hospital admission was 7.8%. Among this admission, anesthesia-related reasons were cited in 44%. CONCLUSIONS: Effective pain management and well-chosen anesthetic technique are important for the success of ambulatory surgery. PMID- 14531267 TI - [The analysis of radial artery pressure waveforms using 4 element model]. AB - BACKGROUND: The pressure wave transmission from the aorta to the peripheral artery has rarely been analyzed using the physiological model linked to the entire vascular system. The goal of the present study is to clarify the law of causality between distortions of waveforms and cardiovascular conditions using a model. METHODS: The arterial system was quantitatively analyzed with a 4-element model consisting of aortic characteristic impedance, inertance, peripheral resistance, and compliance, which shows the nature of second-order response. In order to elucidate the influence of the 4 elements on transmitting waveforms, we simulated cases in which these 4 elements change independently. RESULTS: Large aortic characteristic impedance, low peripheral resistance, and increased compliance cause the damping of radial artery pressure wave. CONCLUSIONS: The model approach enabled us to grasp the hemodynamics-related changes to the pressure waveforms, since each of the 4 elements corresponds to the vascular system with physical meanings. PMID- 14531268 TI - [New blood transfusion system of the Saku Central Hospital--present conditions and problems]. AB - BACKGROUND: It is important for a hospital to have a well organized blood transfusion service. METHODS: Our hospital adopted the maximum surgical blood order schedule (MSBOS) and type and screen (T & S) systems in April 1999 and opened a division of transfusion therapy, thus introducing an integrated control system for blood transfusion. RESULTS: In the year (2002), the crossmatch-to transfusion (C/T) ratio has dropped from 3.94 to 2.01, and the volume of blood transported outside the central operating room and that of waste blood have decreased. CONCLUSIONS: The new systems are instrumental in saving preoperatively prepared blood and exercising its qualitative control. From now on, our task is to improve measures to cope with cases of transfusion in excess of MSBOS and reevaluate MSBOS on a regular basis. PMID- 14531269 TI - [Primary symptoms of 301 patients with prostate cancer: Nagoya Urology Hospital experience]. AB - PURPOSE: Primary symptoms were reviewed retrospectively in patients with prostate cancer diagnosed in our hospital with the aim of promoting early detection. MATERIALS AND METHODS: The subjects included 301 cases with prostate cancer diagnosed histologically in Nagoya Urology Hospital from August, 1988 to December, 2001. The mean age was 72.7 years, and the median PSA was 20.0 ng/ml. Primary symptoms were classified according to the General Rule for Clinical and Pathological Studies on Prostate Cancer (The 3rd Edition). RESULTS: Out of 301 cases, 274 (91%) visited our hospital with clinical symptoms. Of them, 272 had primary urological symptoms. In these 272 cases, 250 (92%) and 19 (7%) had lower urinary tract symptoms (LUTS) and macroscopic or microscopic hematuria, respectively. The majority of patients (82%) referred from other urologists had already undergone PSA measurement, compared to 50% in those referred from physicians other than urologists (p < 0.0005). CONCLUSION: The present data revealed that LUTS were important primary symptoms for the detection of prostate cancer, particularly in an area like Nagoya where the mass screening for prostate cancer is still unavailable. In terms of the early detection of prostate cancer, PSA has to be measured in patients with LUTS even when they visit physicians who are not urologists. PMID- 14531270 TI - [Usefulness of lower ihfundibular length-to-diameter ratio as a predictor of lower pole stone clearance after extracorporeal shock wave lithotripsy; clinical research with two lithotriptors]. AB - PURPOSE: Lower pole renal stones are well known to exhibit a poor stone clearance rate following extracorporeal shock wave lithotripsy (ESWL). In the present study, we analyzed several anatomical factors as predictors of lower pole stone clearance that may be used to indicate the usefulness and the universality of ESWL in such patients with two different lithotriptors. PATIENTS AND METHODS: 93 patients with a unilateral single lower pole stone of 2 cm. or less underwent ESWL using Piezolith 2500 or Medstone STS, were included in the study. An IVP was used to determine the lower infundibulopelvic angle, the caliceal pelvic height, the lower infundibular length, the lower infundibular diameter, the lower infundibular length-to-diameter ratio and the number of lower pole minor calyces. Stone-free status was assessed by a plain film with or without renal ultrasound. RESULTS: The stone clearance rate at the Piezolith 2500 group was 53.1% (34 of 64 patients), while was 51.7% (15 of 29 patients) at the Medstone STS group. In all cases, the overall stone clearance rate was 52.7% (49 of 93 patients). Age, laterality of the stone burden within the kidney and stone size were not different between the stone-free and residual stone groups. Multivariate logistic analysis revealed that length-to-diameter ratio was the most independent predictors of successful stone clearance at each group. The patients exhibited length-to-diameter ratio less than 7 achieved high stone clearance rates, greater than 72%. In contrast, the stone clearance rate was less than one third when length-to-diameter ratio was 7 or greater. Besides length-to-diameter ratio was strong prognostic factor in patients with stones 1 cm. or less and 1 to 2 cm at each group. CONCLUSION: From this study, it is apparent that successful ESWL is highly sensitive to the anatomy of the lower pole of the kidney. Especially, the lower infundibular length-to-diameter ratio is potentially useful and a universal predictor regardless of the kind of lithotriptors at least in patients with a lower pole radiopaque stone 2 cm. or less treated with ESWL. PMID- 14531271 TI - [Clinical evaluation of hot flushes developing during endocrine therapy for prostate carcinoma]. AB - OBJECTIVE: There are few clinical investigations on the hot flushes that develop during endocrine therapy for prostate cancer in Japan, although there are many reports in the Western countries. Therefore, we evaluated the incidence of hot flushes and the association between hot flushes and clinical characteristics of prostate cancer patients receiving endocrine therapy. PATIENTS AND METHODS: Sixty eight prostate cancer patients receiving endocrine therapy (LH-RH analog (group LH-RHA); 22 patients, LH-RHA + non-steroidal antiandrogen (group LH-RHA + NSAA); 20 patients, LH-RHA + steroidal antiandrogen (group LH-RHA + SAA); 8 patients, LH RHA + estramustine phosphate (group LH-RHA + EP); 1 patient, bilateral orchiectomy (group O); 5 patients, O + non-steroidal antiandrogen (group O + NSAA); 11 patients, and O + steroidal antiandrogen (group O + SAA); 1 patient) were evaluated by a fixed questionnaire. The incidence of the hot flush, the association between hot flushes and the clinical factors, as well as the therapy of hot flushes including SAA and Kampo therapy were analyzed. RESULTS: The overall incidence of hot flushes was 37% (36% in group LH-RHA, 45% in group LH RHA + NSAA, 13% in group LH-RHA + SAA, 0% in group LH-RHA + EP, 20% in group O, 45% in group O + NSAA, 100% in group O + SAA). No significant association between the hot flushes and the clinical factors of the patients was observed. On the other hand, in 3 of 4 patients treated by SAA, hot flushes improved after 4 weeks. In 2 of 3 patients treated by Kampo, hot flushes improved after 4 weeks. CONCLUSION: Hot flushes are the major side effect of endocrine therapy for Japanese prostate cancer patients. SAA and Kampo are thought to be effective for treatment of hot flushes. PMID- 14531272 TI - [Primary malignant lymphoma of the prostate: report of a case achieving complete response to combination chemotherapy and review of 22 Japanese cases]. AB - A 70-year-old man presented with complaints of difficult urination, perineal pain and lassitude. An enlarged, hard and nodular prostate was palpable on digital rectal examination. Needle biopsy of the prostate was performed, which revealed diffuse large B-cell non-Hodgkin's lymphoma by immunohistochemical studies. Right internal and external iliac nodes were swollen on computed tomographic scan (CT) of the pelvis. No abnormal finding was seen on abdominal CT, upper gastrointestinal fiberscopy and bone marrow histology. Therefore, the disease was classified into the clinical stage II according to Ann Arbor's criteria. The patient achieved complete response (CR) to five cycles of combination chemotherapy, CHOP, and survives more than two years without recurrence. Primary malignant lymphoma of the prostate is a rare prostatic malignancy. Only 22 Japanese cases with primary prostatic lymphoma have been reported to our knowledge. In 23 cases including ours the majority of the patients were older than 60 years, and their histopathology was mostly diffuse lymphoma, which belongs to intermediate grade of non-Hodjkin's lymphoma according to the Working Formulation's Classification. Nineteen out of 23 cases (83%) were divided into localized stage i.e. stage I or II. In these reports, three of five cases treated with either radical prostatectomy or radiotherapy alone resulted in death or progressive disease. On the other hand, 11 out of 16 cases (69%) who received chemotherapy alone or with other therapy obtained CR. Primary lymphoma of prostate has previously been considered to have a poor prognosis. Our results, however, suggest that patients with this malignancy respond well to combined chemotherapy, and could possibly be cured when the disease is confined to the localized stage. PMID- 14531273 TI - [Extracorporeal shock wave lithotripsy for ureteral stone in patient with implanted cardiac pacemaker: a case report]. AB - We report a case of extracorporeal shock wave lithotripsy (SWL) for ureteral stone in patient with implanted cardiac pacemaker. A 68-year-old woman was admitted to our hospital for left back pain due to left single ureteral stone (13 x 7 mm) in 2002. A permanent cardiac pacemaker has been implanted for sick sinus syndrome in 1997. After evaluation for cardiac function and pacemaker function by a cardiologist and a pacemaker technician, SWL (MFL 5000, Dornier) was performed without changing pacemaker mode (DDD mode). Shock waves were incorrectly exposed a few time triggered by arterial pacing amplitude, but no cardiovascular event or malfunction of the pacemaker was occurred during or after SWL. The ureteral stone was successfully fragmented with 2,400 shock waves (24 kV) and the fragments were delivered immediately. PMID- 14531274 TI - [A case of bilateral neonatal testicular torsion]. AB - We report a case of bilateral synchronous testicular torsion in a newborn. A male neonate was referred one day after birth because his left testis was firm and enlarged. We found that the scrotum was firm bilaterally and suspected it to be bilateral testicular tumor. Surgical exploration which was performed at 2 days after birth revealed testicular torsion on both sides. Left orchiectomy, right detorsion biopsy and orchiopexy were performed. Histopathological examination confirmed hemorrhagic necrosis of bilateral testes. PMID- 14531275 TI - [A case of renal cell carcinoma in tuberous sclerosis]. AB - A 43-year-old woman with mental retardation, epilepsy, and urinary stone disease had a right renal tumor. Acne-like anthema around the nose and dental pits of the nine teeth were typical signs of tuberous sclerosis (TSC), and the biopsy finding of the facial anthema was consistant with TSC. The pathological diagnosis of laparoscopic nephrectomy was renal cell carcinoma in the hemorrhagic cyst. The TSC-related renal cell carcinoma tends to develop bilaterally in younger individuals compared with the sporadic RCC. This case is the 27th case of TSC related RCC in Japan. PMID- 14531276 TI - [Photography of the endoscopic picture with a consumer-grade digital camera]. AB - In spite of the progress of digital camera technology, photography of the endoscopic picture, with a digital still camera, has not spread in the urologic setting. We attempted to photograph an endoscopic picture with a consumer-grade digital camera. An attachment was produced by pasting step-up rings for digital cameras on a coupler, or eye piece, for endoscopes. An endoscope lens, attachment, and the digital camera were connected, and the endoscopic picture was photographed. The endoscopic picture taken with the digital still camera was high definition compared to the one with the video camera for endoscopes. The conditions of the digital camera, which can be used, require having mounting threads for filters on the lens. Although a way of sterilizing and waterproofing a camera must first be discovered, a digital still camera enables one to obtain a high-definition image inexpensively, and to store and arrange the picture. PMID- 14531277 TI - [Protein phosphatases and nucleolin in osteoblastic cells: cleavage of nucleolin in apoptotic cells]. AB - Protein phosphorylation and dephosphorylation has been recognized as a key mechanism in cell proliferation, differentiation, and apoptosis in various tissues. Okadaic acid is a potent inhibitor of protein phosphatase type 1 (PP1) and type 2A and induces apoptosis in human osteoblastic Saos-2 and MG63 cells. Nucleolin is an abundantly expressed nucleolar phosphoprotein and is located mainly in the nucleolus. The staining pattern of nucleolin in Saos-2 and MG63 cells is similar to that of PP1 delta. Nucleolin was demonstrated to bind to PP1 delta in nucleolus by using immunocytochemical and immunoprecipitation methods. AgNORs and nucleolin, visible as dots in the nucleus of the control cells, disappeared from the apoptotic nuclei. A major band, 110 kDa, was detected in the proteins obtained from the control cells. The level of the 110 kDa protein decreased in the apoptotic cells, whereas an additional band, 80 kDa, appeared and the level of this protein increased in the proteins prepared from okadaic acid-induced apoptotic cells. Our results indicate that PP1 delta directly binds to nucleolin in the nucleolus and that nucleolin is cleaved during apoptosis. PMID- 14531278 TI - [Kallikrein-family serine protease in the central nervous system]. AB - Serine proteases exert a variety of functions under physiological and pathological conditions. Tissue plasminogen activator (tPA) is expressed widely in the central nervous system (CNS) and play important roles in development, synaptic plasticity and neuronal cell death. In addition to this protease, recent studies have revealed the existence of new serine proteases in the CNS. In particular, two members of the kallikrein gene family, KLK8/neuropsin and KLK6/protease M/neurosin/zyme are expressed abundantly in the CNS. Neuropsin is expressed by the neurons of the hippocampal subfields CA1 and CA3 and shown to cleave extracellular proteins such as fibronectin and L1. This protease plays essential roles in synaptic plasticity such as long-term potentiation (LTP) and kindling. Application of recombinant neuropsin significantly promoted LTP induction and anti-neuropsin antibody reduced potentiation. Intraventricular administration of anti-neuropsin antibody ameliorated kindling epilepsy. Neuropsin-knockout mice (neuropsin-KO) had significantly smaller number of synapses in the CA1 subfield of the hippocampus. These data suggest that neuropsin plays an important role in synapse formation through modifying extracellular environments. After injury to the CNS, neuropsin is expressed in oligodendrocytes around the lesion. Myelins in the severed optic nerve of neuropsin-KO were more preserved than those of wild-type mice, suggesting that neuropsin after injury is involved in myelin degradation. Another kallikrein member, protease M is constitutively expressed in the oligodendrocytes. Insult to the CNS increases protease M expression not only in the oligodendrocytes but also in the inflammatory cells such as macrophages. These proteases in balance with inhibitors are implicated in the modulation of the extracellular environment. PMID- 14531279 TI - [Enzyme activities and structural features of mitochondria and expression of mRNA of complex I]. AB - The functional ability of the muscle is closely related to the activities of mitochondria, which are the energy producing organelles in the muscle cell. Complex I, III and IV exist as respiratory chain enzymes in the inner membrane of mitochondria and are responsible for transporting electrons from NADH to O2. In this paper, rat masseter muscle, rat and frog cerebral cortex, rat tongue, and frog tibialis anterior muscle mitochondria were described in terms of their correlation with enzyme activity of mitochondrial respiratory chain, the structure and number of mitochondria, and transcription levels of ND1 (mitochondrial) and 51K (nuclear) subunits of Complex I, especially regarding the change in feeding behavior in rat and hibernation in frog. The cristae in mitochondria gradually developed during the postnatal period. The changes in feeding behavior were closely correlated with the level of mitochondrial enzyme activity and were also supported by the structural changes in mitochondria. Based on results of this study, the expression of ND1 and 51K is thought to be involved in the rate-limiting step in the enzyme production. The results also suggest that there is a correlation between biochemical properties and structural features in mitochondria during development, effects of a soft diet, and hibernation. PMID- 14531280 TI - [A model for the embryological instruction of the branchial arteries]. AB - It is difficult to teach students about the embryological transformation of the branchial arteries. In mammals, six pairs of branchial arteries develop, but all are not present at the same embryological stage. The first, second, fifth and a part of right sixth branchial artery disappear early in embryological development. The remaining third, fourth and sixth branchial arteries mainly constitute the arterial system of the breast. The aorta and pulmonary trunk are derived from the truncus arteriosus. Because of the complexity of this developmental process, we have devised a user-friendly model in order to assist with educational presentations. In this model, the shrinkage of a vessel has been represented by inserting a wire inside of the hose representing the artery. Degenerated or disappearing parts of the vessel are removable by using hooks and Velcro tape. Branchial arteries, truncus arteriosus, aortic sac and dorsal arteries are represented by different colors. The descent of the heart is represented by the relational change between larynx and heart. Additionally we represented the vagus nerve and recurrent laryngeal nerve by using strings. The right vagus nerve can move dorsally and the left ventrally by rotating the digestive tract. The right recurrent laryngeal nerve can move superiorly to hook around the right subclavian artery, and left recurrent laryngeal nerve can hook around the ductus arteriosus formed by the left sixth branchial artery. PMID- 14531281 TI - [The application of gene therapy for immunosuppression in allogeneic nerve graft]. PMID- 14531282 TI - [Implantation of cultured urothelial cells into the peritoneal cavity: feasibility of culture condition for seeding cells onto scaffold and histological evaluation of implanted urothelial cells]. PMID- 14531283 TI - [Host gene expression during HIV-1 infection: comparative analysis in human and rat cells]. PMID- 14531284 TI - [A simple and efficient method for constructing an adenoviral cDNA expression library]. PMID- 14531285 TI - [Analysis of TCOF1 gene of eight Japanese patients with Treacher Collins syndrome]. PMID- 14531286 TI - [Donor-derived TNF-alpha can suppress Th1-dependent CTL generation during acute GVHD in mice]. PMID- 14531287 TI - [Characterization of TP53, APC, and PTEN mutational statuses of 120 human cancer cell lines by using yeast-based analyses]. PMID- 14531289 TI - [Long-term management of adult asthma]. PMID- 14531288 TI - [MyD88 is involved in the signaling pathway for paclitaxel-induced apoptosis]. PMID- 14531290 TI - [The roles of eosinophils and lymphocytes in asthma focused on future therapeutic strategies]. PMID- 14531291 TI - [Trend in chemical mediator research in asthma and its application for asthma treatment]. PMID- 14531292 TI - [Molecular mechanisms of allergic airway inflammation in asthma]. PMID- 14531293 TI - [Pathophysiology of airway hyperresponsiveness, the importance of early intervention in the treatment of asthma]. PMID- 14531294 TI - [Mechanisms and control of airway remodeling]. PMID- 14531296 TI - [Impact of cimetidine on growth and metastasis of surgically transplanted Lewis lung cancer]. AB - PURPOSE: Histamine has multiple biological functions and its immunomodulatory actions are not fully understood. In the current study, we investigated the effect of cimetidine, which is a histamine-2 receptor antagonist, on the progress of transplanted Lewis lung cancers using a mouse model. METHODS: A piece of Lewis lung cancer weighing 15 mg was transplanted into the subcutaneous space on the back of each C 57 BL/6 mouse. Mice were randomized into a no-treatment control group (CO) or one of two treatment groups. In the treatment groups, 16 mg/kg/day (LD) or 160 mg/kg/day (HD) of cimetidine was orally administered from one week before the day of transplantation to the time of sacrifice. Subcutaneous tumors and lungs were excised on the 28th or 42nd postoperative day. RESULTS: The mean vascular densities of the subcutaneous tumors on day 28 were 55.7 +/- 23.9/mm2 in CO, 88.0 +/- 16.3/mm2 in LD and 122.6 +/- 16.9/mm2 in HD (p < 0.05; CO vs. LD or HD, LD vs. HD). On day 42, mean weights of the subcutaneous tumors and the numbers of metastatic lung tumors were 6.0 +/- 2.1 g in CO, 7.9 +/- 1.2 g in LD and 10.0 +/- 1.9 g in HD (p < 0.05; CO vs. HD), and 7.5 +/- 6.0 in CO, 17.0 +/- 3.0 in LD and 19.8 +/- 7.4 in HD (p < 0.05; CO vs. HD), respectively. CONCLUSIONS: These results suggest that cimetidine dose-dependently enhances the angiogenesis, growth and metastasis of surgically transplanted Lewis lung cancer in a mouse model of this type. PMID- 14531295 TI - [Coagulation activity in the airways of asthmatic patients]. AB - Fibrin deposition in the mucus plugs of asthmatic patients has long been known, and asthmatic sputum has been held to be important in the pathogenesis of bronchial obstruction. We examined the coagulation activity in the airways of asthmatic patients. Albumin as an index of plasma leakage into the bronchial lumen, thrombin antithrombin III complex (TAT), tissue factor, FDP, D-dimer and the TAT/D-dimer ratio as indices of coagulation and fibrinolytic markers were determined in expectorated or hypertonic saline-induced sputum from patients with acute and stable asthma, and with chronic bronchitis, and from normal control subjects. Patients with acute asthma, in comparison with patients with stable asthma or chronic bronchitis and normal control subjects, had significantly higher levels of albumin, TAT and TAT/D-dimer. The fibrin antigen was more positively stained immunohistochemically in sputum from acute asthmatics than in other sputa. In both patients with acute asthma and those with stable asthma, there was a significant positive correlation between albumin and TAT or albumin and TAT/D-dimer in the sputum. However, in normal control subjects, there was no correlation between these markers. These results suggest that the coagulation system in the airways of acute asthmatic patients is activated, that this favors fibrin deposition in the bronchial lumen and that coagulation pathways in the bronchial compartment and the degree of plasma exudation into the airways are dependently regulated in patients with asthma but not in normal control subjects. PMID- 14531297 TI - [Clinical evaluation of continuous isoproterenol inhalation in adult patients with severe acute asthma]. AB - The clinical efficacy and safety of continuous inhalation therapy using l isoproterenol in adult acute severe asthma were retrospectively evaluated. Fifty adult patients (34 men and 16 women, mean age 47.5) with severe acute asthma, which was not improved by conventional therapy including intermittent inhalation of salbutamol, intravenous aminophylline, and corticosteroid, were given continuous l-isoproterenol inhalation treatment. Both the lowered PaO2/FiO2 and the elevated PaCO2 improved significantly within 4 hours later. Forty-seven patients (94.0%) showed clinical improvement whereas two (4.0%) did not respond and received mechanical ventilation. No obvious side effects were noted in 49 patients (98.0%), while an 81-year-old man showed an elevation of blood pressure, which recovered promptly after discontinuation of the treatment. Based on these observations, we conclude that continuous inhalation therapy using l isoproterenol is a safe and possibly useful option in the treatment of severe adult acute asthma, especially in patients who do not respond to conventional therapy. PMID- 14531298 TI - [A case of pulmonary thromboembolism due to idiopathic thrombosis of inferior vena cava, which was initially misdiagnosed as pneumonia]. AB - We report a case of a 73-year-old man with pulmonary embolism due to idiopathic thrombosis of the inferior vena cava. He was referred to our hospital because of a fever and cough of 2 weeks' duration despite treatment with an oral antibiotic. Chest radiography on the first visit showed an infiltrate in the right middle lung field. He was diagnosed as having pneumonia and admitted to our hospital for treatment. Following administration of intravenous antibiotics, his symptoms disappeared and the chest radiography findings improved. The abdominal CT obtained in an attempt to visualize the cause of liver dysfunction serendipitously revealed thrombosis of the inferior vena cava, which was suspected to have caused the pulmonary embolism. A subsequent lung perfusion scan revealed marked perfusion defects in the right middle and lower lobes. Chest CT revealed an embolus located in the right pulmonary artery. Since thrombolytic therapy was not effective, the placement of a filter in the inferior vena cava was performed to prevent the recurrence of pulmonary embolism. The patient has been asymptomatic without recurrence of the disease since the filter insertion. PMID- 14531299 TI - [A case of primary lung cancer in a young female with tongue cancer]. AB - We describe a case of lung tumor in a 33-year-old woman with tongue cancer. She had noticed a painful sensation in her tongue in April 2000. The results of a physical examination showed a 22 x 11 mm ulcerated lesion on the right side of her tongue. A biopsy specimen showed a moderately differentiated squamous cell carcinoma. A preoperative chest radiograph showed no evidence of pulmonary metastasis. No neck lymphadenopathy was found. The tongue cancer was resected in September 2000, and the pathological stage was T2 N0 M0. A solitary pulmonary lesion appeared 8 months after the surgery. A Histological examination of a transbronchial lung biopsy specimen showed a moderately differentiated squamous cell carcinoma. It was difficult to distinguish histopathologically or immunohistochemically between a second primary tumor and a metastasis, but the pulmonary lesion was considered to be a second primary tumor on the basis of a single endobronchial lesion that appeared after the T2 tongue cancer with no regional recurrence or cervical lymphadenopathy. Treatment with chemotherapy and irradiation was not effective and the patient died 3 months after the diagnosis of lung cancer. PMID- 14531300 TI - [A suspected case of negative PR 3-ANCA Wegener's granulomatosis associated with marked endobronchial lesion and systemic angitis]. AB - A 72-year-old woman had been diagnosed as having organizing pneumonia at another hospital. She was treated with antibiotics, bronchodilators and prednisolone, but her cough, fever and inflammatory signs were not resolved completely. She was admitted to our hospital for further evaluation of the lung lesion. The chest CT scan revealed narrowing of the segmental bronchus and the serum samples showed elevated MPO-ANCA but negative PR 3-ANCA levels. Physical examination also revealed marked bilateral episcleritis and eyeground angitis. Fiberoptic bronchoscopy showed bilateral marked stenotic segmental bronchi associated with mucosal erosions and small nodular lesions. Bronchial and renal biopsy specimens demonstrated marked angitis in the bronchial arteries and glomeruli. A generalized form of Wegener's granulomatosis (WG) was diagnosed and was treated with 60 mg/day of prednisolone, 50 mg/day of cyclophosphamide and 4 g/day of sulfamethoxazole-trimethoprim, which resulted in a marked improvement of the clinical symptoms and endobronchial lesions. In WG, the incidence of endobronchial involvement is 16 to 55% and that of positive serum MPO-ANCA alone is only 14.3%. The present case of WG was considered a rare case with WG which because of the MPO-ANCA-positive but PR 3-ANCA-negative findings associated with marked endobronchial lesions and systemic angitis. PMID- 14531301 TI - [A case of Lemierre syndrome]. AB - We present a case of Lemierre syndrome characterized by thrombophlebitis of the internal jugular vein with multiple metastatic foci after acute otopharyngeal infection in a 30-year-old woman. Despite treatment with tonsillectomy leading to a diagnosis of peritonsillar abscess, her condition worsened and she was admitted with high fever. Chest radiograph and CT scan of the thorax revealed multiple pulmonary cavities and pleural effusion on the right side. On neck CT, a thrombus was detected in the left internal jugular vein. She received with intravenous clindamycin (CLDM) and cefepime (CFPM) and progressively improved. Although Lemierre syndrome is a relatively uncommon disease with the potentially life threatening complication of acute pharyngotonsillitis, this syndrome should be considered in cases of severe tonsillitis or pharyngitis. PMID- 14531302 TI - [A case of allergic granulomatosis and angiitis without symptoms of asthma]. AB - We present a case of allergic rhinitis in a 68-year-old woman in whom eosinophilia was found when she complained of common cold-like symptoms. The patient noticed a mass lesion on her left neck, which improved with antibiotic treatment, but her coughing continued and edema of both lower extremities appeared. She was admitted to our hospital, because of abnormalities in her electrocardiogram and cardiomegaly seen in a chest radiograph. The discomfort due to the edema in the soles of both feet remained even after steroid therapy. Her chest radiograph revealed ground-glass opacity, and a transbronchial lung biopsy revealed granulation tissue with the infiltration of eosinophils into the interstitium. Allergic granulomatosis angiitis was diagnosed because of granulomatosis vasculitis resulting from sural nerve biopsy. This was a rare case of allergic granulomatosis angiitis because her lung function was normal, she had no history of bronchial asthma, and there were no clear symptoms of bronchial asthma. PMID- 14531303 TI - [A case of interstitial pneumonia induced by an ACE inhibitor (temocapril hydrochloride)]. AB - A 51-year-old woman who had been treated for years for rheumatoid arthritis presented with a persistent dry cough and shortness of breath three weeks after administration of the ACE inhibitor temocapril hydrochloride against essential hypertension. Chest radiography and computed tomography showed diffuse reticular shadows and ground-grass opacities in both lung fields. Bronchoalveolar lavage fluid analysis showed an increase of lymphocytes and CD8+ T cells (93.3% of lymphocytes), and a decrease of the CD4/8 ratio of the T cell subset (0.04). Histopathological analysis of trans-bronchial lung biopsy specimens showed infiltration of lymphocytes into the alveolar septa and exudation of alveolar macrophages, signs characteristic of interstitial pneumonia. A drug lymphocyte stimulation test was positive for temocapril, but negative for other drugs. On the basis of these findings, we diagnosed temocapril hydrochloride-induced interstitial pneumonia. PMID- 14531304 TI - [Pulmonary septic emboli in a patient with right-side infectious endocarditis]. AB - An 18-year-old woman afflicted with ventricular septal defect was admitted for high fever and dyspnea. She had undergone no surgical repair. Chest CT showed numerous nodular opacities in both lungs. The majority of them were situated on the pleura. Echocardiography revealed an area of vegetation 20 mm in diameter just beneath the tricuspid valve. Staphylococcus aureus was cultured from venous blood. We diagnosed right-side bacterial infectious endocarditis caused by Staphylococcus aureus and culminating in septic pulmonary emboli. Intravenous panipenem/betamiprom was prescribed, and after 5 weeks, the patient recovered, was negative for C-reactive protein and had a negative venous blood culture. Cardiac septal defect with bacterial endocarditis is a major risk factor in the development of septic pulmonary emboli. PMID- 14531305 TI - [A case of pulmonary carcinosarcoma with abnormal uptake in bone scintigraphy]. AB - A 68-year-old man was admitted to our hospital with abnormal chest radiograph shadows. Chest CT showed a large mass with calcification in the right lower lobe. Bone scintigraphy revealed abnormal uptake by the tumor. The biopsy specimen obtained by bronchoscopy and fine-needle aspiration demonstrated no malignancy, and chest radiographs obtained two years before were normal. Right lower lobectomy was performed. Histologically, the tumor was composed of squamous cell carcinoma and osteosarcoma, and the diagnosis was true carcinosarcoma of the lung. PMID- 14531306 TI - [A case of miliary tuberculosis complicated with a tuberculous aneurysm of the aorta]. AB - A 68-year-old man was admitted because of fever and weight loss. A chest high resolution computed tomography (HRCT) scan revealed diffuse micronodular shadows, and an abdominal CT scan showed an aneurysm spreading from the root of the renal artery to the iliac bifurcation. His fever subsided without treatment and his general condition was good. However, histological studies of a transbronchial lung biopsy specimen and bone marrow aspirate clot revealed non-necrotizing epitheloid granulomas, and mycobacterium tuberculosis was cultured from sputum and bronchoalveolar lavage fluids. Military tuberculosis was diagnosed. One month after the initiation of chemotherapy, the abdominal aneurysm enlarged quickly despite the improvement of the thoracic findings, and graft replacement was performed. Histological findings in specimens of the resected aneurysm suggested that tuberculous inflammation of the surrounding lymph nodes had invaded the aortic wall, leading to the aneurysm. It should be borne in mind that symptoms in elderly patients with miliary tuberculosis may be mild, even when serious extrapulmonary lesions are present. PMID- 14531307 TI - [A case of child obesity hypoventilation syndrome (OHS) improved by bi-level positive airway pressure (BiPAP)]. AB - A 5-year-old boy was admitted to our hospital because of severe obesity and disordered breathing with snoring during sleep. Child OHS was diagnosed using polysomnography (PSG). Although he was treated initially with nasal CPAP, it was not acceptable to him. BiPAP produced marked reduction of the respiratory disorders during sleep, as confirmed by PSG. Few reports of BiPAP for child OHS have appeared in Japan. We concluded that child OHS could be successfully treated with BiPAP when nasal CPAP was not acceptable. PMID- 14531308 TI - [A case of pulmonary Langerhans' cell histiocytosis]. AB - A 42-yr-old woman was referred to our hospital because of multiple small nodules in a chest radiograph. She had no symptoms such as dyspnea, cough or sputum. A chest CT revealed many centrilobular small nodules and thin-walled cysts with predominance in the peripheral area of the lungs. The specimens obtained by thoracoscopic surgery showed granulomas with scattered eosinophils and numerous Langerhans' cells. The Langerhans' cells were positive for both S-100 protein and CD1a. These findings are compatible with pulmonary Langerhans' cell histiocytosis (LCH). Since the granulomas showed no fibrotic changes, the LCH may have been in its early stages. However, there were clusters of lymphocytes and macrophages around the terminal and respiratory bronchioles, and cystic lesions without cellular infiltrates, in the specimens. The former histologic findings suggested respiratory bronchiolitis causing interstitial lung disease and the latter are indistinguishable from centrilobular emphysema. Therefore, these smoking-related diseases may have been superimposed on the LCH in this patient. PMID- 14531309 TI - [Circadian intraocular pressure and glaucoma treatments]. PMID- 14531310 TI - [Effects of ocular hypotensive agents on the circadian rhythm in intraocular pressure in rabbits as measured by telemetry]. AB - PURPOSE: To establish a telemetry system for measuring intraocular pressure (IOP) in rabbits and to evaluate the effects of topical application of ocular hypotensive agents on the circadian rhythm of IOP. SUBJECTS AND METHODS: We developed a telemetry system in rabbits housed under a 12-hour light-dark cycle (light and dark phases: 7:00-19:00, 19:00-7:00, respectively). The IOP resulting from a single topical application of ocular hypotensive agents was measured by telemetry during the light phase and the dark phase. RESULTS: The values measured by the telemetry were positively correlated to the value of the anterior chamber pressure measured by a transducer in range from 5 to 50 mmHg (r = 0.987). A single topical application of timolol maleate (0.5%), dorzolamide hydrochloride (1%), and dipivefrine hydrochloride (0.1%) caused no significant reduction in IOP in the light phase, but they did in the dark phase. A single topical application of bunazosin hydrochloride (0.01% or 0.1%) had significant ocular hypotensive effects in both phases. CONCLUSION: These findings indicate that the different effects of ocular hypotensive agents on circadian rhythms of IOP can be measured by the telemetry. Telemetry may be useful for evaluation of ocular hypotensive agents and the circadian rhythm of IOP. PMID- 14531311 TI - [The influence of hard exudates on the results of vitrectomy for macular edema due to diabetic retinopathy]. AB - PURPOSE: To demonstrate surgical results of diabetic macular edema from the distribution of hard exudates. METHODS: We reviewed 485 eyes of 325 patients followed up more than 1 year who underwent vitreous surgery for diabetic macular edema. Four groups were identified from the distribution of hard exudates in the macular region: no hard exudate type, 179 eyes; hard exudate type, 211 eyes; macular deposit type, 73 eyes; and waxy type, 22 eyes. We evaluated the preoperative characteristics, the period required for absorption of macular edema, visual outcome, and postoperative complications among the four types. RESULTS: In the waxy type, the macular edema was absorbed more slowly than no hard exudate type and hard exudate type(p < 0.05) more patients had proteinurea and were treated by grid photocoagulation before surgery than the other types, and the rate of occurrence of rubeotic glaucoma and attrition by death was higher than in the other types. Visual acuity was significantly improved in all types postoperatively(p < 0.05). Preoperative and postoperative visual acuity were as follows: no hard exudate type, 0.28, 0.48; hard exudate type, 0.21, 0.33; macular deposit type, 0.10, 0.17; and waxy type, 0.04, 0.11. CONCLUSION: The distribution of hard exudates was directly related to surgical results for diabetic macular edema. The waxy type had poor prognosis. PMID- 14531312 TI - [Evaluation of canalization of the canaliculus with digital photo system for treating dry eye]. AB - PURPOSE: We developed a simple evaluation technique of canalization of canaliculus by observing sequential images of the meniscus height after instillation of fluorescein solution. SUBJECTS AND METHODS: We used nine eyes of eight patients [one male eye, eight eyes of seven females, age: 68.3 +/- 15.9 (mean +/- standard deviation)] whose punctal plugs were extruded due to granulation formation within the canaliculi. Fluorescein solution was instilled into the eyes, until the tear meniscus was of sufficient height. Then the tear meniscus at the center of the lower lid margin was photographed with a slit-lamp equipped with a digital camera every three minutes for 15 minutes. These sequential photographs were displayed on the screen of a personal computer and from these photographs, canalization of the canaliculus was evaluated. RESULTS: In six eyes of six subjects, the meniscus height was remarkably diminished with time and the meniscus showed significant decrease as late as 15 minutes. In three eyes of two subjects, no remarkable decrease was noted during the 15 minutes. CONCLUSION: This simple method can be usefully applied in a clinical situation, because canalization of the canaliculus is easily evaluated just looking at the sequential meniscus images on the computer display. PMID- 14531313 TI - [Effect of latanoprost on diurnal variations of intraocular pressure in normal tension glaucoma]. AB - OBJECTIVES: To study the effect of instillation of latanoprost on the diurnal variation in the intraocular pressure(IOP), blood pressure, and pulse rate in patients with normal-tension glaucoma(NTG). SUBJECTS AND METHODS: The diurnal variation in the IOP was determined in 23 eyes in 23 NTG patients after a washout period of 4 weeks or longer. The diurnal variation in IOP was then remeasured after latanoprost monotherapy of 8 weeks or longer. The IOP was measured by the Goldmann applanation tonometer at 10:00, 13:00, 16:00, 19:00, 22:00, 01:00, 03:00, 07:00, and 10:00 before and after treatment, and the IOP at each time point, mean diurnal IOP, maximum IOP, minimum IOP, and range of variation in IOP were compared before and after treatment. The blood pressure and pulse rate before and after treatment were also measured and compared. RESULTS: The IOP decreased significantly at all time points. The treatment caused significant decreases in the mean diurnal IOP, maximum IOP, minimum IOP, and range of variation in IOP. There were no differences in the blood pressure and pulse rate before and after treatment. CONCLUSIONS: Latanoprost significantly decreases the IOP throughout the day in NTG patients, and has no effects on the blood pressure and pulse rate. It is therefore useful in the treatment of NTG. PMID- 14531314 TI - [A case of endogenous Aeromonas hydrophila endophthalmitis]. AB - BACKGROUND: Aeromonas hydrophila(A. hydrophila) may cause septicemia in immunocompromised hosts, but endophthalmitis due to this agent is quite rare. To our knowledge, there have been no reports of A. hydrophila endophthalmitis in Japan. We report a case of endogenous endophthalmitis caused by Aeromonas hydrophila. CASE: The patient was a previously healthy 71-year-old man and no source of the endogenous endophthalmitis was found. A diagnostic/therapeutic vitrectomy was performed, but it was impossible to successfully treat this case because of extended retinal necrosis. A. hydrophila was isolated from the vitreous specimen. CONCLUSION: Endophthalmitis due to A. hydrophila had a rapid clinical course and poor prognosis. PMID- 14531315 TI - [Herpes simplex keratitis after ophthalmic surgery]. AB - PURPOSE: We report 6 cases of herpes simplex keratitis after ophthalmic surgery, in eyes without clinical history of herpes simplex keratitis. CASES: These cases comprised 6 patients examined at our hospital between April 1992 and November 2001. Past operations were keratoplasty in 5 eyes and cataract surgery in 1 eye. Clinical findings and predisposing factors were evaluated retrospectively. The period between herpetic epithelial keratitis onset and ophthalmic surgery ranged from 1.5 to 79 months. Predisposing factors included corticosteroid therapy and operative wound. The herpetic epithelial lesions were dendritic ulcers in 2 eyes, geographic ulcer in 1 eye, and atypical epithelial lesions in 3 eyes; in all cases, herpes simplex virus (HSV)-DNA was detected by polymerase chain reaction (PCR) in tear fluid. All herpetic epithelial lesions healed with oral and topical acyclovir. CONCLUSIONS: When corticosteroids are used following ophthalmic surgery, physicians should be alert to the possibility of herpetic epithelial keratitis, even in patients with no clinical history of herpes simplex keratitis. PCR detection in tear fluid is helpful in diagnosing this disease. PMID- 14531316 TI - Treating drug-abusing offenders. Initial findings from a five-county study on the impact of California's Proposition 36 on the treatment system and patient outcomes. AB - Five counties (Kern, Riverside, Sacramento, San Diego, San Francisco) that demonstrate both variations and similarities in their implementation of Proposition 36 (e.g., treatment approaches, urine testing) and patient mix have been selected to participate in a study assessing how California's Proposition 36 is affecting the drug treatment system and patient outcomes. Except for San Francisco, treatment admissions increased during the first year of Proposition 36 implementation over the prior year (27% in Kern, 21% in Riverside, 17% in Sacramento, and 16% in San Diego), mostly in outpatient drug-free programs. Compared to non-Proposition 36 patients, Proposition 36 patients were more likely to be men, first-time admissions, treated in outpatient drug-free programs, employed full-time, and users of methamphetamine or marijuana. They were less likely to be treated in residential programs or methadone maintenance programs and fewer reported heroin use or injection drug use. Guided by the multilevel open systems framework, the study examines key issues of Proposition 36 that influence treatment systems and outcomes and empirically identifies "best practice" approaches in treating drug-abusing offenders. PMID- 14531317 TI - Descriptive methods for evaluation of state-based intervention programs. AB - In this article, the authors discuss program evaluation of intervention studies when the outcome of interest is collected routinely at equally spaced intervals of time. They illustrate concepts using data from the American Stop Smoking Intervention Study, where the outcome is state per capita tobacco consumption. States differ widely in mean tobacco consumption, and these differences should be accounted for in the analysis. A large difference in the variance of the intervention effect may be obtained depending on whether the variation in the between-state effects are considered. The confidence limits obtained by ignoring between-state effects are too optimistic in many cases. PMID- 14531318 TI - Partners and processes in HIV services for inmates and ex-offenders. Facilitating collaboration and service delivery. AB - The elevated rates of HIV/AIDS among inmates are due in part to the high concentration of incarcerated persons with behavioral risk factors for HIV infection. Corrections, public health, and community-based organizations should partner to take full advantage of the opportunity to maximize effectiveness and efficiency in addressing HIV/AIDS in inmate populations. The purpose of this study is to document issues inhibiting collaboration, service delivery, and multisite evaluation as identified by members of the U.S. Centers for Disease Control and Prevention/Health Resources and Services Administration Corrections Demonstration Project (CDP). Examining the barriers (and identifying ways to resolve them) is important in developing and maintaining successful programs that ultimately benefit HIV-infected inmates and the communities to which they return. PMID- 14531319 TI - Access to breast prostheses via a government-funded service in Victoria, Australia. Experience of women and service providers. AB - For many women, the only alternative to breast reconstruction following a mastectomy is to use external prostheses, which need replacing regularly at a cost of up to $395 per prosthesis. Commonwealth and state governments across Australia have responded to this need by providing subsidies to assist in the purchase of breast prostheses. However, current arrangements have been highly variable and sometimes difficult to access. As part of a larger review of breast prosthesis services in Victoria, Australia, the aim of this research was to evaluate client satisfaction among Victorian women who accessed funds through the State Government's Aids and Equipment Program, compare the responses of the program service providers with the experiences of clients accessing funding, and identify opportunities to improve service provision. PMID- 14531320 TI - [Sexual life after hysterectomy]. AB - OBJECTIVES: The evaluation of sexual life after hysterectomy is still controversial. DESIGN: The aim of the study was to analyse the impact of hysterectomy on frequency and quality of the woman's sexual life. MATERIAL AND METHODS: The total of 539 women operated in II Department of Obstetrics and Gynecology in 1990-2000 were interviewed about symptoms as well as advantages and disadvantages after abdominal hysterectomy. RESULTS: Sexual desire after operation decreased. The rate of dyspareunia dropped. Frequency of sexual intercourse and orgasm decreased non-significantly. The percentage of women non reporting vaginal dryness decreased significantly from 18.5% to 37.3% after hysterectomy. CONCLUSION: Sexual activity overall improved after hysterectomy. PMID- 14531321 TI - [Prognostic value of the pronuclear morphology pattern of zygotes for implantation rate]. AB - INTRODUCTION: In ICSI and conventional IVF cycles, the embryos are usually selected for transfer based on evaluation of the cleavage speed and the extent of blastomere fragmentation. Pronuclear stage scoring system has been used in IVF recently as an additional parameter. OBJECTIVES: The aim of this study was to classify embryos from ICSI cycles according to the pronuclear scoring system and its influence on implantation rate. MATERIAL AND METHODS: 62 couples suffering from different types of infertility and treated by ICSI were included in our study. For each embryo, the following data were analyzed: pronuclei (PN) morphology, 16-20 hours after ICSI, and embryo morphology, about 40-42 (day 2) and 65-68 (day 3) hours after ICSI. The embryo transfer was performed on the third day after pick-up. RESULTS: In 55 (88.7%) cases we transferred at least one embryo from the zygotes of 0 or 1 or 2 pattern. The implantation rate was 30.6% (n = 19). In all of these cases the embryos have been derived from zygotes from pattern 0, 1 or 2. CONCLUSIONS: Pronuclear scoring system of zygotes appears to be a predictor of the implantation potential. Embryos derived from pattern 0, 1 and 2 of zygotes implant at a higher rate (p = 0.05). PMID- 14531322 TI - [Evaluation of the advantages of laparoscopic procedures for hysterectomy]. AB - OBJECTIVE: The objective of this publication was the analysis of 102 laparoscopic hysterectomy. The results were discussed in comparison with traditional laparotomy. MATERIAL AND METHODS: There was the analysis of 102 hysterectomy by the means of laparoscopy done in 2000-2002. Following parameters were evaluated: duration of the operation, blood loss, complication, the day of introducing enteral nutrition and duration of hospitalization. RESULTS: It was observed, that laparoscopic operation took more time than laparotomy (average time: 96.4 min vs 62.37 min). Blood loss was similar in both procedures. Comparing the changes of hemoglobin concentration in serum before and after the procedures it was observed average change 1.75 g% for laparoscopy and 1.71 g% for laparotomy. Time introducing enteral nutrition and duration of hospitalization was shorter for laparoscopic procedures. 87.25% of patients was able to eat in next day after laparoscopic operation. Average amount of days of hospitalization after laparoscopic procedures was 3.75 days, after laparotomy 6.44 days. The infections were the most common postoperative complications. The infections were more common in laparotomy group. CONCLUSIONS: The new method causes: shortened time of hospitalization, quicker introducing of general diet, avoiding the large wound of abdominal segments, small amount of complications. PMID- 14531323 TI - [Laparoscopic management during pregnancy--own material (1996-2002)]. AB - OBJECTIVE: Utility of laparoscopic procedures in pregnant women. Is laparoscopy safe for the foetus? MATERIAL AND METHODS: Between 1996 an 2002 14 pregnant patients were admitted and laparoscopy was performed. RESULTS: In 10 cases laparoscopic cyst enucleation (6 simplex, 1 endometrial, 3 dermoidal), in 1 case myomectomy and in 2 adnexectomy was performed. There were no complications during surgery. In one case miscarriage occurred 3 weeks after laparoscopic surgery and 9 healthy babies were delivered. DISCUSSION: There is no doubt that laparoscopy is well accepted in gynecological surgery and is going to be more and more accepted in obstetrics. Due to several differences between non-pregnant and pregnant women laparoscopic procedures in pregnancy require special attention. All papers show good foetal outcome even with preterm labour. In our material all babies were born healthy. CONCLUSIONS: Laparoscopic procedures during pregnancy are feasible and safe. Nevertheless they have to be performed by experienced gynecological-surgical teams. PMID- 14531324 TI - [Cutaneous melanoma during pregnancy at the Cancer Center-Institute in Warsaw]. AB - Malignant melanoma is a neoplasm which frequency has been increasing rapidly in Poland. The paper describes five cases of patients with cutaneous melanoma coexisting with pregnancy, who have been recently treated in the Department of Soft Tissue/Bone Tumors of M. Sklodowska-Curie Memorial Cancer--Institute of Oncology, Warsaw. All patients presented clinically advanced primary lesions with poor prognosis. In paper recommendations for treatment of women with suspicious cutaneous lesions before or during pregnancy and patients with melanoma planning a pregnancy are presented. The suspicion of melanoma is indication for immediate excisional biopsy by local anesthesia. Further treatment should be performed in tertiary oncological centres. There is no evident clinical data that pregnancy has a significant impact on progression of melanoma. The most important factor influencing prognosis of melanoma is the stage of the disease at the diagnosis. There are no grounds for abortion in pregnant women with melanoma in I/II/III stages. PMID- 14531325 TI - [Influence of folic acid in primary prevention of neural tube defects]. AB - OBJECTIVE: The aim of the study was to analyze the incidence of neural tube defects in Lublin Province between 1990 and 2000 after application of primary prevention with folic acid. MATERIAL AND METHODS: A total of 293,039 newborns were analyzed. RESULTS: Neural tube defects were diagnosed in 53 newborns. CONCLUSION: Reduction of frequency of meningocele was shown after application of primary prevention with folic acid in 1998. Meningoceles were most frequent in 1992 (1.32/1000) and in 1998 frequency of these defects was reduced and in 2000 was down to 0.62/1000 births. PMID- 14531326 TI - [Tissue and blood oxygenation in newborns < 1250g and incidence of ROP]. AB - AIM: The aim of this study was to analyze correlations between the onset and severity of retinopathy and premature birth. MATERIAL AND METHODS: In the three groups of 30 newborns < 1250 g we have analyzed blood saturation, tissue oxygenation and blood transfusion. To estimate tissue oxygenation we used following formulas; 1. TVCE = [formula: see text] V-blood volume = 8% of body weight 2. Oxygen availability = (0.54 + 0.005 x gestational age) x Hb (g/dL) Additionally we investigated incidence of BPD, PDA, severe IVH, and sepsis. RESULTS: We have found statistically significant influence of drops of SaO2 < 85% and lower mean values of SaO2 on development of severe retinopathy. We have not seen any statistical differences in the other parameters of oxygenation between the groups. CONCLUSIONS: Fluctuation as well as instability of oxygen blood saturation seem to be the most important risk factors of retinopathy. Limitation of early blood transfusions can decrease incidence of ROP. We do not have adequate methods for the estimation of tissue oxygenation which can be used to verify indications for blood transfusion. We should use very restrictive recommendations for blood transfusion in ELBW babies. PMID- 14531327 TI - [Cardiac arrest and implantation of a cardioverter-defibrillator in a pregnant woman]. AB - During pregnancy increased incidence of maternal cardiac arrhythmias is observed. We reported a 31 year old primipara after two incidents of cardiac arrest due to ventricular fibrillation during pregnancy at 10th and 16th week of gestation. The implantable cardioverter defibrillator was implanted at the 19th week of gestation. There was no complication after ICD implantation. A healthy child was born by cesarean section at 38th week of gestation. PMID- 14531328 TI - [Pregnancy presence in residual uterine horn]. AB - The development of the 20-th week pregnancy in residual uterine horn was observed in 25 years old women in first pregnancy. The pregnant women was hospitalised in the Department of Obstetrics and Gynaecology Medical University in Lodz. The acute stomach pain was the only symptom of disease. Ultrasound examination shows the presence of 20-th alive, normal pregnancy and the presence of adjunctive uterus. The patient was operated due to the signs of hemorrhage. The rupture of the residual horn and the presence of the pregnancy in the abdominal cavity was found. The postoperative status was normal. PMID- 14531329 TI - [Burkitt lymphoma involving the ovaries]. AB - Burkitt lymphoma is a very rare type of lymphoma with a predilection for the ovary. We report a case of 18-years old woman with dramatic course of the Burkitt lymphoma involving ovaries. Immunohistochemistry techniques allowed for the diagnosis and the successful chemotherapy. The case demonstrates the importance of an appropriate pathology evaluation and interdisciplinary collaboration. PMID- 14531330 TI - [Hereditary ovarian cancer as a separate clinical entity]. AB - Family history of ovarian cancer has been regarded as the most important risk factor in developing this type of tumor. Recent identification of cloning of specific tumor suppressor genes responsible for hereditary ovarian cancer syndromes, resulted in the surge of enthusiasm and optimism regarding the practical application of genetic information. Many investigators believe that natural history of hereditary ovarian may significantly differ from sporadic cases. Some independent findings have shown that serous adenocarcinoma was the most frequent histological type in hereditary ovarian cancer patients, whereas the mucinous and the borderline were underrepresented in this group. Also, patients with hereditary ovarian cancer tend to develop the disease at younger than expected age. At the present moment, it remains unknown if environmental/lifestyle factors play a role in penetrance and expression of specific tumor suppressor genes in mutation carriers. The era of molecular genetics has raised hopes not only for a better understanding of biology and natural history of hereditary ovarian tumors but also a rapid breakthrough in prevention and management of this disease. PMID- 14531331 TI - [Chances in current prenatal diagnosis]. AB - Authors present the actual trends in prenatal imaging diagnoses: screening obstetrical ultrasound (performed in every ultrasound lab), genetic (targeted) US + fetal echocardiography (performed in selected ultrasound lab), US 3D and 4D, panoramic studies, surgical US, fetal MR and social US study. Authors also present opinion about who, where and how should perform counseling after detection of fetal anomaly. Also current trends in obstetrical education are presented based on examples from West Europe and USA. PMID- 14531332 TI - [Ductus venosus blood flow in normal pregnancy outcome]. AB - OBJECTIVE: Doppler studies of the fetal circulation have demonstrated the importance of ductus venosus velocimetry in the monitoring abnormal course of pregnancy. DESIGN: Estimation of the ductus venosus Doppler reference ranges between 10-19th week of pregnancy. MATERIAL AND METHODS: The ductus venosus blood flow has been studied in 140 consecutive fetuses between 10 and 19 week of pregnancy. When a midsagittal view of the fetus was obtained, color Doppler was used to visualize the ductus venosus. The pulsed Doppler sample volume was then placed on it, in the isthmic portion. RESULTS: The mean PIV ranged from 1.09 in the first trimester of pregnancy to 0.96 in the second trimester. CONCLUSION: A decrease in mean PIV values was observed in the course of pregnancy. In the normal course of pregnancy A-wave was observed. PMID- 14531333 TI - [Surgical complications in patients treated for invasive cervical cancer]. AB - OBJECTIVES: The aim of this study was to analyse, retrospectively, complications of therapy of 898 patients suffered from invasive cervical carcinoma. All patients have undergone radical surgical treatment in Gynaecological Department of Medical University of Gdansk between 1972 and 2000. MATERIALS AND METHODS: Patients were divided into four groups according to clinical examination (FIGO staging from 1985). Group Ia consisted of 27 (3%) patients; Ib consisted of 711 (79%) patients; IIa consisted of 133 (15%) patients and IIb consisted of 27 (3%) patients. 85 (9.5%) patients ware older than 60. In all cases radical abdominal hysterectomy and pelvic lymph nodes dissection was applied. RESULTS: Intraoperative complications ware observed in 66 (7.3%) patients. Among them 2 (0.2%) patients ware died, 29 (3.2%) patients had massive hemorrhage and 37 (4.1%) patients had local organs damage as bladder, rectum or ureter wall. During the postoperative period 11 (1.2%) patients ware died. There ware observed ureterovaginal fistula in 25 (2.7%) cases and vesicovaginal fistula in 9 (1.0%) cases. CONCLUSION: Radical surgical treatment of invasive cervical carcinoma performed in our department is relatively safe procedure. Results showed above are similar to the results from leading oncology centers. PMID- 14531334 TI - [Laparoscopy in the diagnosis and treatment of pathologic lesions of the genitourinary tract]. AB - OBJECTIVE: The objective of this article is retrospective analysis of laparoscopic surgery that were performed from January 2002 to December 2001 in I Ward of Gynaecology and Obstetrics of Gynaecology and Obstetrics Hospital in Warsaw. MATERIAL AND METHOD: 464 laparoscopies that were performed from January 2000 till December 2002 were analysed. Evaluation was made on the basis of: indications for surgery, kind of surgery procedure that was performed, blood loss during surgery, complications and hospitalisation period. RESULTS: The most common indication (39.86%) was non-malignant tumors of cases. Another pathologies of cases: ectopic pregnancy, endometriosis and infertility were respectively 11.21%, 3.02% and 10.34% cases diagnosed before surgery. Many procedures that have been done recently by laparotomy are now performed by laparoscopy, that is: enucleation of a cyst of adnexa, removal of an ectopic pregnancy, hysterectomy. The blood loss during surgery was evaluated on the basis of change of haemoglobin concentration in serum before and after surgery. The average blood loss was 1.2 g%. There were 39 cases of conversion of laparoscopy to laparotomy. The most common reason of conversion (16 cases out of 39) was insufficient hemostasis in the operative area or bleeding in the wound after trocar. We had noted the small amount of infections (4.5%). The average hospitalisation period after laparoscopy was 2.62 days. CONCLUSIONS: Laparoscopic procedures can be widely used in gynaecology, have the small number of complications, short hospitalisation period and allow to avoid the big postoperative wound of abdominal segments. PMID- 14531335 TI - [Oral contraceptive practices among female medical students of the University of Warsaw]. AB - DESIGN: Apparently no researches have been conducted regarding oral contraceptive usage among female medical students in Poland. In the last ten years, all over the world, only five publications have dealt with this topic. OBJECTIVES: The purpose of the research reported in this article was to establish the types, choices and frequency of usage of oral contraceptive pills among female medical students of the University of Warsaw. MATERIAL AND METHODS: 634 female students responded to a specially designed questionnaire. RESULTS: Data collected indicated that, 52.2% of the students have been involved in sexual intercourse. 167 of them used Oral Contraceptive Pills (OCP). Two third survey participants used OCP following a doctors recommendations. CONCLUSIONS: 1.30.5% of the sexually active students used OCP. 2.31.9% of this group of students believed the use of OCP was the most effective of all the contraceptive methods. 3.65.0% of the respondents used OCP following a doctors recommendations. 22.7% of the survey participants used OCP in order to regulate their menstrual cycles. 4. The respondents who reported customary use of OCP cited the most frequently used OCP as: Cilest-28.0%, Mercilon-19.2% and Minulet-13.2%. PMID- 14531336 TI - [Influence of valproic acid (depakine I.V.) on human placenta metabolism- experimental model]. AB - OBJECTIVES: The pregnancy in women with epilepsy is associated with an increased incidence of congenital malformations in offspring. Currently, anti-epileptic drugs (AEDs) are concerned to be a major etiologic factor of abnormal fetal development but the pathomechanism of teratogenicity of AEDs is complex and not well understood. The purpose of this study was to evaluate an influence of one of the AED-valproic acid (VPA) on placental metabolism (glucose consumption and lactate production). MATERIAL AND METHODS: Term human placental cotyledons were perfused in vitro using a recycling perfusion of maternal and fetal circulations. A total 18 placentas were perfused either with 75 micrograms/ml of VPA (therapeutic dose) or with 225 micrograms/ml of VPA (toxic dose). Eight placentas were perfused with a medium without VPA and served as controls. During 2.5 h of experiment, both maternal and fetal glucose consumption and lactate production were measured every 30 minutes. RESULTS: The introduction of different concentrations of VPA into the perfusion system did not effect placental glucose consumption and lactate production rates in both maternal and fetal compartments. CONCLUSIONS: The teratogenic effect of valproic acid is not associated with metabolic disturbances of glucose or lactate in the placental tissue. PMID- 14531337 TI - [Course of pregnancy, delivery in adolescents treated in I Clinical Unit of Obstetrics and Gynecology in Bytom (1997-2001)]. AB - OBJECTIVES: The subject of our analysis was pregnancy, delivery and perinatal outcome in adolescent girls treated in I Clinical Unit of Obstetrics and Gynecology in Bytom in the period 1997-2001. MATERIAL AND METHODS: Our materials were 164 pregnant adolescent girls aged 14-18. Course of pregnancy, type of delivery, perinatal outcome and the recourse to cesarean section were retrospectively analysed in our study. RESULTS: Mean age of adolescent patients in this study was 16 years and 10 months. 141 pregnancies were conducted by natural birth and 23 (14%) by cesarean section. The rate of preterm birth in this analysis was 15.85%. CONCLUSIONS: 1. Adolescent patient pregnancy need special perinatal care. 2. The perinatal outcome of analysed cases were similar to other authors. PMID- 14531338 TI - [Assessment of pregnancy and labour outcome and the condition of the newborn in primiparous women aged 35 and older]. AB - OBJECTIVE: To assess the effect of maternally advanced age on pregnancy and labour outcome. MATERIAL AND METHODS: Pregnancies of 95 primiparous women aged 35 or older were compared with pregnancies of 130 women aged 20-30 with respect to pregnancy complications and labour outcome. RESULTS: Women aged 35 or older had cesarean section three times as often as women in the 20-30 year group old. Advanced maternal age compared with younger age was associated with a significantly higher rate of diabetes, hypertensive disorder, thrombocytopenia and intrauterine growth retardation. CONCLUSIONS: Primiparous women aged 35 or older are at an increased risk of complications in pregnancy and are at a higher risk of cesarean section compared with younger women. PMID- 14531339 TI - [Nitric oxide activity in women with intrauterine growth restriction treated by L arginine]. AB - OBJECTIVES: The aim of the study was to find correlation between the nitric oxide metabolites such nitrite in healthy pregnant women and in intrauterine growth restriction. The decrease of nitric oxide metabolites production may be linked to endothelial cells deficiency. DESIGN: High risk of destruction of biological function of cells by free radical processes is observed in IUGR. It depends on NO deficiency and lipid peroxidation processes. Concentration of NO metabolites measured in pregnancy can show the correlation between oxidative stress and NO metabolism. MATERIAL AND METHODS: The study was undertaken in Medical University in Lodz in 2000-2002. The study group consisted of women with IUGR, the control one of healthy women. The group of IUGR was treated with L-arginine-nitric oxide precursor and acetylsalicylic acid. The nitrite activity was measured by Oxis kit, in Grees reaction and was expressed in mumol/l. RESULTS: In group of IUGR low concentration of NO metabolites was observed before treatment. After 20 days of treatment the concentration decreased. In the control group the value was higher than IUGR group/significant difference/and after 20 days of observation decreased as well. CONCLUSIONS: We found the difference between the two groups and the decrease of NO metabolite concentration in the course of pregnancy. PMID- 14531340 TI - [Maternal and umbilical bilirubin concentration at the time of delivery depending on course of pregnancy and labour]. AB - OBJECTIVES: Newborns hyperbilirubinemia is an interesting, complicated and controversial clinical problem. An elevated concentration of it can be connected with increased risk of jaundice. Also, maternal bilirubin can play a part in the development of jaundice in mature newborns. MATERIAL AND METHODS: The research included a group of 173 healthy, full-term newborns in generally good condition, with body mass between 5 and 95 percentile. Infants were born in 2002 at 1st Department of Obstetrics and Gynaecology Medical University in Warsaw. The maternal blood was collected during labour and the umbilical blood was collected immediately after delivery. RESULTS: The mean value of maternal bilirubin was 0.62 mg/dl, and of total bilirubin in the umbilical blood was 1.61 mg/dl. CONCLUSION: The differences between respective values in children delivered by caesarean section, whose mothers suffered from the infection during pregnancy or narcotic analgetic drugs that had been administered during labour were not statistically significant. Maternal bilirubin was higher in mothers with proteins < 6.0 g/dl. The umbilical bilirubin was statistically increased in children whose mothers oxytocin received. PMID- 14531341 TI - [Need for surgical ligation of patent ductus arteriosus among patients treated at the Neonatology Department of the University of Medical Sciences, Poznan, Poland]. AB - OBJECTIVE: Clinical assessment of newborns requiring surgical ligation of patent ductus arteriosus (PDA). MATERIAL AND METHOD: 51 newborns operated on PDA in Department of Neonatology in Poznan in years 2000 and 2001. Newborns were classified for surgical ligation after echocardiographic examination (ACUSON 128 XP/10 or HDI 3500, sector scanhead 5MHz). Following data were analyzed: antenatal steroid therapy, gestational age, birth weight, Apgar score, the reason of respiratory insufficiency after birth, duration of mechanical ventilation and oxygen therapy, incidence of bronchopulmonary dysplasia (BPD), intraventricular haemorrhage (IVH) and deaths. RESULTS: 30 of 51 analyzed patients had very low birth weight (< or = 1000 g). Antenatal steroids were used in 21 cases, 20 newborns were delivered by caesarean section. The status of 22 newborns was assessed after birth as bad (0-3 points in Apgar scale). In 38 newborns respiratory distress syndrome was diagnosed and 32 required surfactant therapy. Surgical ligation of PDA was performed on 19.8 +/- 14.4 day of life in neonates born in our hospital and on 24.3 +/- 11.9 day of life in newborns admitted for ligation from other wards. Surgery of one patient was complicated by pneumothorax. 42 patients required mechanical ventilation before operation. Mean duration of mechanical ventilation was 33.4 +/- 26.9 days. 30 infants developed BPD and 15 infants IVH III or IV degree, 6 infants died. CONCLUSIONS: PDA is a serious clinical problem especially for very low birth weight newborns. Surgical ligation of a PDA, without transportation of a patient, is associated with a low incidence of complications. PMID- 14531342 TI - [The course of a multipara's pregnancy, labour and puerperium complicated by spleen tumor]]. AB - DESIGN: The multipara with spleen's tumor was diagnosed and observed during pregnancy, labour and puerperium. The spleen's tumors are very rare and they should be examined individually. Termination of the pregnancy by cesarean section was performed because of the big risk for the mother and child (tumor's crack, haemorrhage, infection). After 3 months post partum the spleen with tumor was operated (splenectomy). It was cystic spuria. RESULTS: The cystic spleen was diagnosed by accidental abdominal USG and this test prevented threatening complications. CONCLUSIONS: The abdominal USG should be used in pregnancy. PMID- 14531343 TI - [Wernicke's encephalopathy due to hyperemesis gravidarum]. AB - A report of 33 years old female with Wernicke's encephalopathy is presented. The disease was secondary to hyperemesis gravidarum, started from the 6th week of pregnancy. Neurological symptoms as nystagmus, headache, vertigo, disturbance of consciousness and ataxia are described, as well as difficulties in finding out the right diagnosis. We present the therapy with vitamin B1. The subsequent course of the pregnancy was uncomplicated and resulted in a birth--by caesarean section of a healthy male infant, weighted 2790 grams. However, four months later after the delivery, the patient is not yet completely recovered and still demands thiamine supplementation. PMID- 14531344 TI - [Strategies for fertility preservation after anti-cancer therapy]. AB - A tremendous advance in the field of cancer therapy resulted in a substantial improvement of long-term survival of oncological patients. The most frequent side effect of either chemotherapy or radiotherapy is a partial or complete lost of fertility potential. This paper reviews current knowledge on the fertility preservation strategies for patients facing oncological problems and the risks of iatrogenic infertility. The cryopreservation of spermatozoa or testicular tissue combined with assisted reproductive technology is the optimal solution for male patients. In the case of female patients, the use of ovarian transposition, ovarian suppression and ovarian tissue cryopreservation seem to be the valuable options for fertility prevention. The cryopreservation of embryos is another possible solution, although this procedure is only limited to the couples. On the other hand, there are still controversies regarding the ethical issues concerning gametes and embryos banking. PMID- 14531345 TI - [The data of Polish Vital Statistics Office are correct]. AB - OBJECTIVES: Perinatal mortality rate is important expression of public health care. Estimation of the real effects of medical procedures needs a proficient information flow. MATERIAL AND METHODS: Authors compared data from Polish Official Statistics with data transferred from obstetric departments of regional hospitals in Lublin province. RESULTS: There were significant divergences concerning neonatal mortality. Hospitals reported 33 deaths less than statistical office. In order to explain the divergence found the new forms for reporting were prepared. CONCLUSION: Secondary analysis of the results exerted, that data obtained from Polish Official Statistics were correct. PMID- 14531346 TI - [The relationship between intrapartum fetal heart rate disturbances, electrocardiographic changes and umbilical venous troponin-I (T-I)]. AB - OBJECTIVE: To analyse the relationship between intrapartum fetal heart rate disturbances and electrocardiographic changes and umbilical venous troponin-I (T I), concentrations as well. MATERIAL AND METHODS: 14 fetuses were continuously surveyed by CTG and ECG recordings in the first and second stage of labor, using STAN S-21 analyser. After birth, umbilical venous blood samples were collected for determination of acid-base balance, base excess and troponin-I concentrations. RESULTS: From among monitored fetuses, in 8 cases the CTG and FECG patterns were correct. Vaginally delivered neonates were born in good clinical status, with normal acid-base balance, base excess and T-I < 0.3 ng/ml. In 3 cases abnormal CTG patterns were observed with early decelerations but FECG patterns were correct. Vaginally delivered neonates were born in good clinical status, with normal acid-base balance, base excess and T-I < 0.3 ng/ml. In 2 cases abnormal CTG patterns were observed with variable decelerations but FECG patterns were correct. Vaginally delivered neonates were born in good clinical status, with normal acid-base balance, base excess and T-I < 0.3 ng/ml. In one case abnormal CTG pattern were observed with late decelerations but FECG patterns was correct. The pregnancy was terminated by caesarean section because of fetal distress. The neonate was born in good clinical status with normal acid-base balance and base excess but T-I concentration was increased (1.5 ng/ml). PMID- 14531347 TI - [Chronic obstructive pulmonary disease--an overlooked life style disease]. PMID- 14531348 TI - [Chronic obstructive pulmonary disease. Admission, course and prognosis]. AB - INTRODUCTION: The aim of this study was to describe the prognosis after hospitalization after exacerbation of chronic obstructive pulmonary disease (COPD). MATERIAL AND METHODS: A retrospective study of 300 consecutively admitted patients with exacerbation of COPD in three departments of internal medicine with special interest in respiratory medicine. Data were collected from patient charts. RESULTS: The mean age was 71.3 years. 59.7% were women and 60.9% had cor pulmonale. In 44% of the patients hypoxia was documented at the time of admission. Twentysix(9%) patients died during hospitalization. After three and 12 months mortality was 19% and 36%, respectively. Age, cor pulmonale and hypoxia at the time of admission were associated with increased mortality, whereas impaired lung function, long term oxygen treatment, and a record of hospitalization with the same diagnosis in the previous 30 days were associated with increased mortality in univariate analysis, but not when adjusted for age, gender, and hypoxia at admission. The risk of readmission was increased in cor pulmonale, while cessation of smoking during hospitalization reduced this risk. DISCUSSION: Patients who have been hospitalized with exacerbation of COPD has a high mortality and a high risk of readmission. PMID- 14531349 TI - [Imatinib--a breakthrough in the treatment of gastrointestinal stromal tumors (GIST)]. AB - Gastrointestinal stromal tumours (GISTs) are rare gastro-intestinal, mesenchymal tumours characterized by the expression of a receptor with tyrosine kinase activity called c-kit. A new drug, Imatinib, is a potent inhibitor of a subgroup of the tyrosine kinase family comprising BCR-ABL, platelet-derived growth factor, and c-kit. Imatinib represents the first systemic treatment with a clinical effect on patients with metastatic or unresectable GISTs, which are known to be resistant to chemo and radiotherapy. In the first phase I and II studies confirmed partial responses were seen in 53% and 59% of the patients respectively. PMID- 14531350 TI - [FDG-PET and CT in gastrointestinal stromal tumor treated with imatinib]. AB - The combined imaging modality PET/CT is a possible superior tool for evaluation of gastro-intestinal stromal tumor treatment illustrated in a case of a 59-year old woman responding to imatinib. PMID- 14531351 TI - [Research conditions at the preclinical medical, odontological and pharmaceutical fields]. AB - The Danish Medical Research Council has recently published a report (in Danish) entitled "Research Conditions in the Pre-clinical and Dental Institutes of the Health Science Faculties and the Danish University of Pharmaceutical Sciences". According to this report, the Danish educational institutions within health sciences are facing a major generational change among the scientific staff within the coming 5-15 years. It is therefore worrying that young physicians, dentists and pharmacists refrain from choosing a university research career. Possible explanations include more attractive working conditions, salaries, and career possibilities outside universities. The analyses describe a number of problems regarding this matter and outline how the situation might be improved. The Council recommends that to ensure a future optimal staff balance, focus should be placed on recruitment of researchers. It is important that the future scientific staff also includes a substantial number who possess the specific education provided by the faculty. The staff should be made up of a mix of researchers with knowledge about the nature and practice of their specific discipline and researchers with other scientific backgrounds in the health sciences. It is this interplay between the basic and clinical disciplines that must be consolidated in order to meet the future challenges. The interdisciplinary nature of Danish health science has undoubtedly contributed to placing the Danish health care services in the international front line. PMID- 14531352 TI - [Crisis of the scientific communication]. AB - Scientific communication is undergoing radical changes. The increasingly high prices of scientific journals and other barriers limiting their accessibility are important reasons. On the internet, it is possible to archive and publicize articles with open access, thereby speeding up scientific communication and achievements. Thousands of researchers around the world and many institutions are in favour of open access, while commercial publishers fear losing their profits. The new open-access journals have a low impact factor, which is impending the transformation of the publication system. PMID- 14531353 TI - [Adrenal scintigraphy]. AB - Adrenal scintigraphy is a functional imaging technique for evaluation of adrenal gland hyperfunction. In this review the methodology and multiple applications of the examination are described. The role of scintigraphy in the light of the relatively high radiation dose to the patient is defined and compared with other imaging modalities. The request for a thorough biochemical and anatomical evaluation prior to scintigraphy is emphasized in order to achieve accurate interpretation. Abnormal imaging patterns and the interpretation criterion are discussed. PMID- 14531354 TI - [Late sequelae after severe intracranial hemorrhage in 23 premature children. Parental perspectives]. AB - INTRODUCTION: Cerebral hemorrhage in very premature infants results in high mortality and increases the risk of handicap. The aim of the study was to evaluate: 1) the parental opinion of the child's motor function, 2) their opinion of the child's development compared to their expectations, and 3) how they experienced the information given by nurses and doctors at the time of the diagnosis. MATERIAL AND METHODS: Parents of 23 of 24 premature infants answered a questionnaire. During the neonatal period all infants had suffered from a major cerebral hemorrhage. The children had reached an age between two and seven years. RESULTS: Half of the children had motor deficits (48%), and 22% of the children had severe motor problems. Future development and motor function were reliably estimated by the doctors during the neonatal period. Most of the parents (79%), however, estimated that their child's development was better than expected. The majority of the parents were satisfied with the information that had been given at the time of diagnosis, and they had received adequate support. DISCUSSION: Cerebral hemorrhage in premature infants is still a major problem. Our results, however, do not suggest major change of clinical practice. PMID- 14531355 TI - [The prevalence of skin and mucosal symptoms in gardeners handling Ficus benjamina (weeping fig) and Hedera helix (ivy). A cross-sectional study]. AB - INTRODUCTION: Allergic and toxic initiative symptoms from skin, eyes and respiratory tract are well known among gardeners This study reports the prevalence of these symptoms among gardeners working with Ficus Benjamina (Fb) and Hedera helix (Hh). MATERIAL AND METHODS: A questionnaire concerning allergic/toxic-irritative symptoms from skin, eyes and respiratory tract were forwarded to 57 gardeners. A total of 49 questionnaires were returned and matched on sex and age with a control group. A number of 46 women and 52 men participated, with a mean age of 39 (SD 9.7). Differences in frequencies of symptoms were expressed as odds ratio (OR) with a security level of 95% after conditioned logistic regression; other results were calculated from Mantel Haenzel test. RESULTS: There were more complaints among gardeners compared to the control group f as to symptoms of asthmatic bronchitis (OR 4.6 [0.98-21.2]), rhino conjunctivitis (OR 6.4 [2.1-20.2]) and skin symptoms (OR 4.3 [1.5-12.5]). The symptoms were especially profound among the caretakers of Fb and Hh and to a lesser degree among gardeners working in greenhouses growing Fb and Hh. The symptoms of the gardeners were to a higher degree independent of the season (OR 3.1 [1.1-9]), related to working environment (OR 16.1 [3.3-78.9]) and indoor work (OR 24 [4.8-118.7]). DISCUSSION: This study seems to demonstrate high frequencies of apparently allergic and/or toxic irritative symptoms among gardeners and should be followed by a study including clinical examinations. With knowledge of the allergenic and toxic irritative properties of Fb and Hh it seems problematic that these plants are found in such high numbers in public and private places. Allergic testing including these plants should be performed to a higher degree than is actually the case. PMID- 14531356 TI - [Effect on endometrium of long term treatment with continuous combined oestrogen progestogen replacement therapy. A follow up study]. PMID- 14531357 TI - [Picture of the month: recurrent pancreatitis]. PMID- 14531358 TI - [What is the cost of albumin transfusion in undernourished patients?]. PMID- 14531359 TI - [Does "commercial bias" exist?]. PMID- 14531360 TI - [Antidepressive agents: overtreatment or undertreatment?]. PMID- 14531361 TI - [The preventive treatment of depression]. PMID- 14531362 TI - [Weeping tour in Washington D.C]. PMID- 14531363 TI - [The God created obesity!]. PMID- 14531364 TI - [Concerning educational positions and shortage of specialists]. PMID- 14531365 TI - [The cost of voluntariness. A comment to the AIDS strategy--15 years after "Fall of the law"]. PMID- 14531366 TI - [Sarcopenia--age-related loss of muscular mass and muscular strength]. PMID- 14531367 TI - [Sarcopenia and strength training. Age-related changes: effect of strength training]. AB - Sarcopenia signifies the age-related loss of muscle mass and consequently muscle strength. Sarcopenia appears to be caused by both muscular and neural factors. Concurrently with the muscle atrophy, a non-linear loss of muscle strength is observed. The decline accelerates after the age of 60. The ability to produce muscular power is reduced even more than the muscle strength. Strength training increases muscle strength and muscular power in the elderly thus counteracting part of the age-related reduction. Improvements, however, depend on the initial strength in the elderly person. The benefit of strength training is greatest in frail elderly and the oldest old, although elderly in general could benefit from strength training. Considering the growing section of elderly in the population, the focus on sarcopenia and measures to counteract this seems more relevant than ever. PMID- 14531368 TI - [Severe acute respiratory syndrome--SARS]. AB - Severe Acute Respiratory Syndrome (SARS) is an acute respiratory illness caused by SARS coronavirus. This virus was possibly transmitted from an animal reservoir to humans, and from February 2003, the epidemic was spread internationally by further person-to-person transmission. The SARS epidemic was managed by well known principles of infection control, including prompt diagnosis, isolation of patients, and quarantine of contacts. The successful control of the outbreak is a remarkable international achievement, though much about SARS remains poorly understood. PMID- 14531369 TI - [Socioeconomic risk indicators for disability pension within the Danish workforce. A registry-based cohort study of the period 1994-1998]. AB - INTRODUCTION: The aim was to identify socio-economic risk indicators among the Danish work force (aged 18 to 59) for being granted disability pension. MATERIAL AND METHODS: A closed cohort study based on a random 10% sample of the Danish workforce defined in 1994 and followed in registers from 1995 to 98 for disability pension. The study population included 254,905 persons. Citizens in Denmark have a unique personal identification number so socio-economic data at individual levels from Statistics Denmark were linked with the Disability Pension Registry. All persons but 39 could be traced throughout the study or until the event of disability pension, emigration or death. Cox regression was used to calculate the hazard rate ratio of being granted disability pension. RESULTS: 4443 persons were granted disability pension. The risk of being granted disability pension increased significantly with low social class. Comparing unskilled workers to managers showed that workers had 1.6 (95% confidence interval: 1.4-1.9) times greater risk of disability pension. For female unskilled workers the risk was 1.9 (1.6-2.2). Men with seven years' schooling had 3.1 (2.3 4.3) times higher risk of disability pension than men with a university degree, and women with seven years' schooling had 3.8 (2.5-5.8) greater risk. Having been on sick leave more than 13 weeks compared to sick leave less than four weeks increased the risk by 11.5 (10.1-13.1) for men and 11.2 (9.9-12.8) for women. High age, unemployment, living alone and receiving social benefits were also risk indicators for disability pension. DISCUSSION: The study identified reliable estimates of socioeconomic risk indicators, e.g. low level of education, low social class, and unemployment for leaving the workforce with disability pension. PMID- 14531370 TI - [Surgery patients' intake of herbal preparations and dietary supplements]. AB - INTRODUCTION: Herbal medicine is being frequently used by patients around the world. Several products may interact with ordinary medicine, so it is important for doctors to know what kind of herbal medicines their patients take. MATERIAL AND METHODS: A questionnaire was given to presurgery patients during a two-month period. RESULTS: A total of 115 consecutive patients responded, 69 women (60%) and 46 men (40%). 50.4% had taken or still took herbal medicine, with the following distribution of gender: women 69.8%, men 30.2%. The age group was 18-82 years. The frequently used herbal medicines were fish oil, ginkgo, Echinacea, Co Q10, garlic, and hip. Twenty-five patients took nutritional supplements with the following spread of gender: 84% women and 16% men. The frequently used nutritional supplements were Gerimax, LongoVital, and Melbrosia. Not all patients would inform their doctor about their use of herbal medicine. 28.6% retained information because of the doctor and 64.3%, did not perceive herbal medicine as "real medicine". DISCUSSION: It is important for anaesthesiologists to know what specific kind of herbal medicines patients are using before they anaesthetize them as interactions between herbal medicine and anaesthesia are prevalent. Anaesthesiologists have to ask specific questions to receive full information regarding herbal medicines during the preoperative period. PMID- 14531371 TI - [Metabolic disease or shaken baby syndrome?]. AB - We describe two children with subdural haematoma and glutaricacidaemia type 1, who were diagnosed late because of initial suspicion of shaken baby syndrome. PMID- 14531372 TI - [Cardiac peptides--seen from the basic point of view]. PMID- 14531373 TI - [The surgical service in Denmark--time for reorganization?]. PMID- 14531374 TI - [Increasing drug costs]. PMID- 14531375 TI - [Physician's role in sick leave: an idiot, a prostitute or just a hostage?]. PMID- 14531376 TI - [Health costs, traffic--and the new editorial guidelines]. PMID- 14531377 TI - [Outlook on evidence]. PMID- 14531378 TI - Best of the American College of Cardiology 52nd annual session. March 30-April 2, 2003, Chicago, Illinois, USA. PMID- 14531379 TI - Severe acute respiratory syndrome: Relapse? Hospital infection? PMID- 14531380 TI - Remembering Jonathan M. Mann in a world ajar. PMID- 14531381 TI - Mild severe acute respiratory syndrome. PMID- 14531382 TI - Transmission of severe acute respiratory syndrome. PMID- 14531383 TI - Home-prepared hamburger and sporadic hemolytic uremic syndrome, Argentina. PMID- 14531384 TI - Q fever in Thailand. PMID- 14531385 TI - World Health Organization global conference on severe acute respiratory syndrome. PMID- 14531386 TI - Vincent van Gogh (1853-1890). The Prison Courtyard (1890). PMID- 14531387 TI - Comparative Clinical and Molecular Endocrinology. Proceedings of a symposium. Zeist, The Netherlands, 30-31 August 2001. PMID- 14531388 TI - [Chemical Contamination of the Environment: Risk Assessment. Proceedings of the General Polish Educational Conference. May 8-9, 2003, Pultusk]. PMID- 14531389 TI - Contrast media in magnetic resonance angiography. PMID- 14531390 TI - Widening of the greater trochanteric physis in the immature skeleton: a radiographic sign of femoral osteomyelitis (2003:6b). PMID- 14531391 TI - Abstracts of the 50th British Society for the Study of Prosthetic Dentistry Annual Conference, April 2003. PMID- 14531393 TI - Signalling the Future. Proceedings of an independent meeting. Liverpool, United Kingdom, 4-6 September 2002. PMID- 14531392 TI - The 1971 smallpox outbreak in the Soviet city of Aralsk: Implications for Variola virus as a bioterrorist threat. Proceedings of special issue. PMID- 14531394 TI - AMPK 2002. Proceedings of the 2nd International Meeting on AMP-activated Protein Kinase. Dundee, Scotland, 12-14 September 2002. PMID- 14531395 TI - To biopsy or not to biopsy. PMID- 14531396 TI - [Proceedings of the 4th Congress of Cystic Fibrosis, from Childhood to Adult. Lille, France, 27-28 March 2003]. PMID- 14531398 TI - Borrowed buildings: Canada's temporary hospitals during World War I. AB - This article explores the alterations and additions made to houses, colleges, and larger public buildings in their temporary use as convalescent homes for wounded Canadian soldiers during World War I. These ''borrowed buildings'' anticipated and emphasized three important characteristics of subsequent hospital architecture by suggesting that domestic environments enhanced healing, that hospitals should be accessible, and that patients should be segregated according to social status and disease. The broader implications of the study are that temporarily appropriated spaces contribute to the definition of many institutions and often act as arenas of resistance to established practices. PMID- 14531399 TI - From home to hospital: Parallels in birthing and dying in twentieth-century Canada. AB - This article highlights the striking parallels in the history of birthing and dying as they moved from home to hospital in Canada in the twentieth century. It shows that deaths, like birth, has been hidden in hospitals only since mid century. It demonstrates that the availability of paid and unpaid female caregivers was a key factor affecting the shifts in location of death, much as it was for birth. Thus, women have been the major consumers and caregivers of health services at both ends of the life cycle. PMID- 14531402 TI - George Fulford and Victorian patent medicine men: Quack mercenaries or Smilesian entrepreneurs? AB - In the early twentieth century, a time when patent medicine men were stereotyped as evil and dishonest, G. T. Fulford of Brockville, Ontario made his fortune from an iron pill called Dr. Williams' Pink Pills for Pale People. Once successful, Fulford remained in Brockville where he served on the town council and gave generously to charities. In 1900 he was appointed by Laurier to the Senate. When he died in 1905 he was remembered as a kind and ethical man. His story, like that of several other prominent patent medicine men, conforms more with the ideals of Samuel Smiles than with the popular image of disrepute. PMID- 14531403 TI - Challenges posed by health care restructuring in Ontario. PMID- 14531404 TI - Establishing standards for health care archives: a case study of the Calgary Regional Health Authority. AB - Due to recent restructuring of Alberta's health care system, many historically valuable records are at risk. However, Freedom and Information and Protection of Privacy legislation has created awareness of the need to care for both administrative and medical records of health care institutions, and standards for the maintenance of these records are evolving. The Calgary Regional Health Authority has established archives, records management, and access and privacy programs, and now has the opportunity to establish a serious medical research repository. The CRHA's experience proves the feasibility of preserving historical health care records for both medical and historical use. PMID- 14531405 TI - Review of Christopher J. Rutty, a circle of care. PMID- 14531406 TI - The successful consultant's skill set. PMID- 14531407 TI - Urinary albumin excretion predicts cardiovascular and non-cardiovascular mortality in general population. PMID- 14531408 TI - Search-based optimization. AB - The problem of determining the minimum cost hypothetical ancestral sequences for a given cladogram is known to be NP-complete (Wang and Jiang, 1994). Traditionally, point estimations of hypothetical ancestral sequences have been used to gain heuristic, upper bounds on cladogram cost. These include procedures with such diverse approaches as non-additive optimization of multiple sequence alignment, direct optimization (Wheeler, 1996), and fixed-state character optimization (Wheeler, 1999). A method is proposed here which, by extending fixed state character optimization, replaces the estimation process with a search. This form of optimization examines a diversity of potential state solutions for cost efficient hypothetical ancestral sequences and can result in greatly more parsimonious cladograms. Additionally, such an approach can be applied to other NP-complete phylogenetic optimization problems such as genomic break-point analysis. PMID- 14531409 TI - Reconstruction after total gastrectomy. PMID- 14531410 TI - Injury of the ovarian vein during appendectomy. PMID- 14531412 TI - 2003 IASH Lifetime Achievement Award. Dr Carlos Ferrario. PMID- 14531413 TI - 29th annual North American Neuro-Ophthalmology Meeting, Snowbird, Utah, February 8-13, 2003. PMID- 14531414 TI - How radiosensitive are the developing embryo and fetus? PMID- 14531415 TI - Anaerobic digestion of organic solid wastes: process behaviour in transient conditions. AB - This paper presents the study of the variations of the process parameters during transient conditions when substrates with different biodegradability were fed to a pilot-scale anaerobic digester. The two substrates used in the experiments were the mechanically selected organic fraction of municipal solid wastes and a blend of mechanically selected and separately collected organic fraction of municipal solid wastes. Two transient conditions were studied: the mesophilic-thermophilic passage and the increase of the organic loading rate in thermophilic conditions. It was shown that enhanced fractions of biodegradable substrates determined a greater perturbation of the system and needed longer time to reach a new stable condition. The volatile fatty acids concentration and their indirect measure, alkalinity determined upon pH 4, was the more useful parameter to understand the process behaviour because of its sensitivity to the process variations. PMID- 14531416 TI - On-line NIR monitoring during anaerobic treatment of municipal solid waste. AB - An anaerobic digester (81) was fed with the organic fraction of municipal solid waste and monitored intermittently for two years with on-line near-infrared (NIR) spectroscopy and traditional chemical parameters analysed off-line. The dynamics that occurred due to changes in substrate composition (changed C:N ratio) and changes in operating conditions (overloading) could be followed using principal component analysis of the obtained NIR-spectra. In addition, process disturbances such as failed stirring and increased foaming were readily detected by the NIR spectra. Using PLS regression the propionate concentration could be predicted in the range 0.1-3.6 g/l, RMSEP 0.53 g/l with slope 0.74 and correlation coefficient 0.85. The response on changes in the digester fluid was reproducible and could be detected within 2.5 minutes, which can be considered as real-time monitoring. PMID- 14531417 TI - Comparison of dry and wet digestion for solid waste. AB - To reduce the amount of MSW going to landfills a number of technologies have been developed. Two main types of anaerobic digestion processes are distinguished for MSW organic waste, which are generally referred to as "wet" (10-15% DM) and "dry" (24-40% DM) anaerobic digestion processes. The input is conditioned to the appropriate DM content by adding process water as required. This article compares a full-scale dry process, Valorga in La Coruna, Spain and a wet process, Vagron in Groningen, The Netherlands. A crucial difference is that the Vagron facility applies a washing step prior to the AD process to remove inert materials for re use and to prevent damage of the installation. In the Valorga facility the organic fraction of the mechanical separation process is fed directly to the AD process. Both processes can be considered proven technology. Specific gas production is practically identical. Waste water production is higher in the wet process, as is to be expected, but this is compensated by a smaller amount of digestate to be disposed of and the separation of inert materials suitable for recycling. The organic loading rate for the Vagron process appears to be higher and the required reactor volume smaller in comparison to the dry Valorga process. The applicability of AD processes is strongly determined by the environmental standards set for the products from digestion. The German standards for digestate result in a lower potential for recycling of inert material separated from organic MSW. PMID- 14531419 TI - Process enhancement in anaerobic bioreactor landfills. AB - The development of bioreactor landfills originated with the recognition that the sequential acid and methane fermentation phases of anaerobic waste stabilization could be accelerated by leachate recirculation. Original studies suggested the potential for such a change in design and management of landfills receiving both municipal and industrial solid wastes, followed by both pilot- and full-scale bioreactor landfill demonstrations. The enhancement of waste transformation within controlled bioreactor landfills is addressed in terms of the sequential phases of waste stabilization, temporal and spatial distribution of leachate and gas generation patterns, mechanisms of conversion, and proposed process and operational modifications. Selected results from investigations on bioreactor landfills include co-treatment of organic and inorganic constituents and the use of dedicated treatment zones. PMID- 14531418 TI - Comparison of the biodegradability of the grey fraction of municipal solid waste of Barcelona in mesophilic and thermophilic conditions. AB - The results of the start-up of two digesters in mesophilic and thermophilic conditions, together with its steady results at several organic loading rates are described. A kinetic study is also carried out which allows one to estimate the ultimate methane production, together with the first-order kinetic constant. Operation at thermophilic temperature yields better results as it allows a more loaded reactor and the methane production is slightly higher. PMID- 14531420 TI - Evaluation of the current status of operating and closed landfills in Russia, Finland and Ireland with regard to water pollution and methane emission. AB - The annual production of municipal solid wastes (MSW) in Russia, Finland and Ireland in the late 1990s accounts for 37.5, 2.5 and 2.05 min. tonnes or 252, 488 and 566 kg per capita, respectively. 96.5, 64 and 91% of these wastes (for Russia, Finland and Ireland, correspondingly) are currently disposed of via landfilling. However, nowadays, MSW management in these countries is undergoing drastic changes (source separation, closure of old landfills, reduction of the number of landfills etc.) forced by recent legislation set by the European Union and Russian authorities. This paper evaluates the current status of MSW landfills, as well as information on current leachate and methane emissions in the three above mentioned countries. Landfill leachates are highly variable in each country and between different countries due to different rainfall and climatic conditions and also due to poor landfill top insulation/cover. Leachates in poorly structured landfills are very dilute, whereas leachates with total COD and nitrogen contents as high as 33,700 mg COD/l and 4,030 mg N/l, respectively, have been detected from state-of-the-art sites. Currently, on-site treatment of leachates exists at only a few landfills in Russia, Finland and Ireland but this situation will be considerably improved during the next years. The annual methane emissions from landfills are estimated as 500-900 and 77 ktonnes for Russia and Finland, respectively. Recent estimates from Ireland suggest an annual landfill methane emission of c. 2.1 Mt CO2 equivalent. Several systems of methane recovery have been developed in all three countries and these are currently in different stages of implementation. PMID- 14531421 TI - The analysis of methods for measurement of methane oxidation in landfills. AB - Landfills and dumps are important sources of atmospheric methane. There is no generally accepted estimate of the influence of methane oxidation on landfill methane emissions. The present work aimed to analyse different methods for the investigation of methane emission and oxidation in methane-producing environments (wetlands, landfills, sludge checks), and to develop the precise procedure for the landfills. The combination of geochemical and microbiological methods to estimate and monitor the oxidation and emission of methane in landfills during different seasons is proposed. It includes the measurements, both on the surface and at different depths (up to 1 m) of landfill ground of the following parameters: (1) concentrations of methane, carbon dioxide and oxygen; (2) quantity of 13C isotope in gas samples; (3) methane-oxidation activity of landfill grounds assayed with two different methods: (a) in conditions of no moisture or substrate limitations, and (b) in conditions with a minimal deviation to in situ conditions; (4) the density of methanotrophic microbial population. PMID- 14531422 TI - Enhancement of polycyclic aromatic hydrocarbons removal during anaerobic treatment of urban sludge. AB - Anaerobically stabilized sewage sludge has potential to partially substitute synthetic fertilizers. The main risk with the recycling of urban sludge on agricultural soils is the accumulation of unwanted products, such as trace metals and organic micropollutants. In this context, the polycyclic aromatic hydrocarbons (PAHs) are particularly monitored because of their toxic properties at low concentrations and their high resistance to biological degradation. The aim of the present study was to optimize PAHs removal during anaerobic digestion of contaminated sewage sludge. Thirteen PAHs were monitored in laboratory-scale anaerobic bioreactors under mesophilic (35 degrees C) and thermophilic (55 degrees C) methanogenic conditions. Abiotic losses were statistically significant for the lightest PAHs, such as fluorene, phenanthrene and anthracene. It was shown that PAH removal was due to a specific biological activity. Biological PAHs removal was significantly enhanced by an increase of the temperature from 35 degrees C to 55 degrees C, especially for the heaviest PAHs. Bioaugmentation experiment was also performed by addition of a PAH-adapted bacterial consortium to a non-acclimated reactor. Significant enhancement of PAHs removal was observed. It was finally shown that PAH removal efficiencies and methanogenic performances were closely linked. The rate of biogas production may be used as an indicator of bacterial activity on PAH removal. PMID- 14531423 TI - Anaerobic digestion of waste activated sludge combined with ozone post-treatment and recycling. AB - The aim of the study was to determine the performances of a combined ozone/anaerobic digestion system for waste activated sludge reduction. The objective was the estimation of the process efficiency and stability when keeping constant influent flow while increasing recycled chemically treated flow. The ozonation step consisted in a partial oxidation (0.16 g O3/g SS) of the anaerobic mesophilic digested sludge. Chemical treatment of digested sludge resulted in a threefold COD solubilization and a decrease of SS of 22%. Some of the advantages of digested sludge ozonation were: deodorization, better settlement and a reduction in viscosity. However there were drawbacks: foaming during ozonation and, at high ozone doses, poorer filterability. The anaerobic digestion was carried out over 6 months with an increasing recycling of ozonated flow. Suspended solids removal rate and COD removal rate were compared with initial operating conditions for the biological reactor and the whole combined process. The optimum recycling rate was 25% with increases of SS removal and COD removal of 54% and 66% respectively when considering the combined process; corresponding to a decrease of the hydraulic retention time from 24 days to 19 days. PMID- 14531424 TI - High-rate anaerobic hydrolysis and acidogenesis of sewage sludge in a modified upflow reactor. AB - Continuous experiments were conducted to study the hydrolysis and acidogenesis of sewage sludge in an upflow reactor with an agitator and a gas-liquid-solid separator. Results of this study showed that 34-78% of volatile suspended solids (VSS) in sewage sludge was hydrolyzed at pH in the range 4.0-6.5, 35 degrees C and 4-24 hours of hydraulic retention time (HRT). About 31-65% of carbohydrate in sewage sludge, 20-45% of protein and 14-24% of lipid were acidified in this reactor. Hydrogen production was favored in lower pH and HRT, whereas methane production was encouraged at higher pH and HRT. Acetate, propionate, butyrate, and i-butyrate were the main aqueous acidogenic products. The distribution of these compounds in the effluent was more sensitive to pH, but was less sensitive to HRT. The maximu specific COD solubilization rate and specific volatile fatty acids production rate were 126 mg-COD/g-VSS x d and 102 mg-VFAIg-VSS x d, respectively. Compared with a CSTR, this modified upflow reactor was shown to be a more promising biosystem for the hydrolysis and acidogenesis of sewage sludge. PMID- 14531425 TI - Alkaline subcritical-water treatment and alkaline heat treatment for the increase in biodegradability of newsprint waste. AB - This work describes two alkaline semicontinuous processes for the conversion of refractory organic materials into biodegradable substances. Newsprint was used as a lignocellulosic waste. Methane conversion efficiencies and cellulose removals were investigated for the two following processes: alkaline subcritical-water treatment (ASWT) coupled with methane fermentation and alkaline heat treatment (newsprint heated with steam in an autoclave; AHT) coupled with methane fermentation with a neutral subcritical-water treatment (NSWT) recycle. Results showed that for ASWT coupled with methane fermentation higher methane conversion efficiencies and higher cellulose removals were achieved as HRT increased. At HRT = 20 days, average CH4 conversion efficiency and average cellulose removal reached 26% and 44%, respectively. After a final HRT of 40 days, average CH4 conversion efficiency and average cellulose removal reached 50% and 60%, respectively. On the other hand, for AHT coupled with methane fermentation, methane conversion efficiencies did not show a greater improvement using this pretreatment process. Average conversion reached 9% with an average cellulose removal of 20%. In order to improve the yield of the reactor, approximately one third of the effluent was recycled using NSWT (150 degrees C; neutral pH). Methane conversion efficiency of this process increased as more recycles were performed. For the fifth operation, the total average methane conversion efficiency was 44% with a total average cellulose removal of 55%. PMID- 14531426 TI - Optimal process configuration for anaerobic digestion with ozonation. AB - Economical source minimization of excess sludge production is an attractive option to deal with the problem of sludge disposal under strict disposal standards. In this paper long-term operational results for two different process configurations that combine oxidative ozone treatment with anaerobic sludge digestion are described. In the first configuration ozone pretreatment was combined with chemostat anaerobic digestion while in the second configuration ozone pre/post-treatments were combined with an anaerobic digester operated without solid removal. From the results of chemostat experiments, the ozone pretreatment solubilized around 19% and 37% of the solids at 0.015 and 0.05 gO3/gTS ozone dose respectively. The ozone pretreatment resulted in improved TVS reduction efficiencies and the degradation efficiencies were observed to depend on the applied ozone dose and system SRT. The TVS degradation efficiency for pre ozonated sludge at an ozone dose of 0.05 gO3/gTS was 59% as compared to 31% for the control reactor fed with un-ozonated sludge. Test results with the second configuration indicated that overall TVS removal efficiencies for a process scheme with post-ozonation could be improved up to 85% with a minimum ozone dose of 0.045 gO3/gTVS-fed. However, since no solids (except that for sampling) were withdrawn in this configuration, the accumulated total solids in the reactors increased to 28 g/l to 30 g/l at pseudosteady state. The average specific methane recoveries were observed to be 0.36 lCH4/gTVS fed which were slightly lower than theoretically expected. Based on the experimental results, important points in the choice of process configuration are discussed. PMID- 14531427 TI - Using quantitative real-time PCR to determine the hygienic status of cattle manure. AB - We developed a suitable system of DNA extraction and real-time quantitative polymerase chain reaction (qPCR) for the specific and sensitive quantification of pathogens and other relevant (indicator) organisms in recalcitrant material such as cattle manure. PCR inhibition by coextraction of humic compounds was minimized in this system, resulting in detection sensitivity of one target DNA copy per reaction well. Data from qPCR analysis for Escherichia coli agreed with cultivation based results, but orders of magnitude more fecal enterococci, Enterobacteriaceae and Campylobacter jejuni, were determined by qPCR than by cultivation. These bacteria may have been in a potentially hazardous active but non-cultivable state. The qPCR system is much less time consuming than conventional cultivation, highly specific, can detect non-cultivable organisms, provides high measurement throughput, and is cost attractive. It should be considered as an alternative in various application areas for (prescribed routine) cultivation based assays, e.g. for biosafety and hygiene monitoring. PMID- 14531428 TI - Investigation of variations in microbial diversity in anaerobic reactors treating landfill leachate. AB - A young landfill leachate containing elevated total ammonia concentrations frequently as high as 2,700 mg/l was anaerobically treated for 1,015 days in two different laboratory-scale anaerobic reactors configured as sludge blanket and hybrid bed. In this paper, the last 265 days of this long-term anaerobic treatability study are presented. Effects of high ammonia concentrations on reactor performances were correlated to the variations in microbial diversity by identifying the dominant microorganisms with FISH (fluorescent in-situ hybridization), cloning, DGGE (denaturing gradient gel electrophoresis) and morphological analysis. The results have indicated that the high ammonia landfill leachate can be treated successfully by using either an UASB or a hybrid bed reactor if temporary pH adjustments in the reactor influents are made when high ammonia concentrations are experienced. Consequently, COD removal efficiency is independent of microbial diversity and reactor configuration and it depends on the biodegradable portion of the leachate. Under this circumstance, stability of the reactors with low levels of acetate was supported by the abundance of Methanosaeta population. In both of the reactors, some populations of Methanobacteriaceae were also detected while other methanogenic species were virtually absent. However, after the termination of pH adjustment at Day 860, reactors immediately became unstable due to the sudden increase in free ammonia concentration up to 400 mg/l. COD removal efficiency decreased to 42% in the hybrid bed and to 48% in the UASB reactor. The durations of inhibitions were not long enough to severely deteriorate the massive Methanosaeta cells; therefore, many of them were again identified after two free ammonia inhibitions. However, subsequently, long filamentous morphologies of Methanosaeta cells shifted to shorter filaments and they lost their aggregating property. PMID- 14531429 TI - Population dynamics of rumen microbes using modern techniques in rumen enhanced solid incubation. AB - The microbial ecology of the rumen is very complex. Different species of bacteria, protozoa, and fungi are involved in digestion of plant material in ruminants. In spite of complicated interrelationships among the various groups of microorganisms in the rumen ecosystem, Bacteria and Archaea are believed to play a major role because of their numerical predominance and metabolic diversity. In this work we are presenting the results for microbial population dynamics of rumen microbes during two-stage anaerobic digestion of grass. The reactors were inoculated with fresh rumen content. Fluorescent in situ hybridization, confocal laser scanning microscopy and epifluorescence microscopy were employed for microbial investigation. It was observed that Bacteria dominated in the hydrolytic reactor (1st stage) whereas Archaea were predominant in the methanogenic reactor (2nd stage). The stability of the methanogenic reactor was result of the dominance of Methanosaeta species (mainly the filamentous type). PMID- 14531430 TI - Co-digestion of concentrated black water and kitchen refuse in an accumulation system within the DESAR (decentralized sanitation and reuse) concept. AB - Co-digestion of concentrated black water and kitchen refuse within the DESAR concept was the objective of this pilot research. The digestion took place in two, non-mixed accumulation reactors (AC1 and AC2) inoculated with digested primary sludge from a WWTP at a temperature of 20 degrees C for a period of around 150 days. Reactor AC1 was fed with a mixture of faeces, urine and kitchen refuse in the equivalent amount that one individual generates per day. The AC2 was fed with a mixture of faeces and kitchen refuse in the equivalent amount that two individuals produce per day. Some contribution of urine to AC2 was not to be avoided. Detailed characterisation of waste(water) was performed. The performance of the stratified reactor was followed by monitoring the reactor content for several reactors' heights as well as being based on the biogas production. In general the system exposed good process stability. The methanisation of 34 and 61% was obtained for AC1 and AC2 respectively. The biogas yield was 26.5 and 50.8 L/p/d for the respective reactors. Proper choice of inoculum as well as good buffering capacity did not lead to accumulation of VFA and an inhibitive effect due to relatively high ammonium concentration. The chosen process is a promising technology showing good process stability especially for high strength influent. PMID- 14531432 TI - Impacts of separate rejection water treatment on the overall plant performance. AB - A separate rejection water treatment appears as a high-tech unit process which might be recommendable only for specific cases of an upgrading of an existing wastewater treatment plant. It is not the issue of this paper to consider a specific separate treatment process itself but to investigate the influence of such a process on the overall plant performance. A plant-wide model has been applied as an innovative tool to evaluate effects of the implemented sidestream strategy on the mainstream treatment. The model has been developed in the SIMBA environment and combines acknowledged mathematical descriptions of the activated sludge process (ASM1) and the anaerobic mesophilic digestion (Siegrist model). The model's calibration and validation was based on data from 5 years of operating experience of a full-scale rejection water treatment. The impact on the total N-elimination efficiency is demonstrated by detailed nitrogen mass flow schemes including the interactions between the wastewater and the sludge lane. Additionally limiting conditions due to dynamic N-return loads are displayed by the model's state variables. PMID- 14531431 TI - Co-digestion of primary sewage sludge and industrial wastewater under anaerobic sulphate reducing conditions: enzymatic profiles in a recycling sludge bed reactor. AB - The first stage in the degradation and recycling of primary sewage sludge and particulate organic matter is the solubilisation and enhanced hydrolysis of complex polymeric organic carbon structures associated with the anaerobic sulphidogenic environment. Solubilization of complex carbon substrates provides the primary reaction in the BioSURE Process, and is effected in the recycling sludge bed reactor (RSBR). During the process of anaerobic digestion, macromolecules are broken down into simpler low molecular weight compounds in the presence of extracellular enzymes. Though hydrolysis of the complex carbon sources was enhanced under biosulphidogenic conditions, no studies have examined the role of enzymes and the enzymatic profiles in the RSBR. To investigate the overall enzymology in the RSBR variations in COD, pH, sulphate, sulphite and sulphide concentrations, carbohydrates, protein and activities of glucosidases, proteases and lipases were studied over 50 days in the reactor at three different depths: 0-16 cm; 16-32 cm; 32-50 cm. While the pH profile remained fairly constant between 6.9 and 7.3 the sulphate and sulphide levels, as expected, changed dramatically as sulphate reduction took place. Proteases, lipases and glucosidases all showed enhanced activity with depth in the bioreactor. It is suggested that the increased sulphide concentration generated during the sulphate reduction process stimulates the enzymes, leading to enhanced solubilisation of primary sewage sludge. PMID- 14531433 TI - A distributed model of solid waste anaerobic digestion: sensitivity analysis. AB - A distributed model of anaerobic digestion of solid waste was developed to describe the balance between the rates of polymer hydrolysis and methanogenesis during the anaerobic conversion of rich and lean wastes in batch and continuous flow reactors. Waste, volatile fatty acids (VFAs), methanogenic biomass and sodium concentrations are the model variables. Diffusion and advection of VFAs inhibiting both polymer hydrolysis and methanogenesis were considered. A sensitivity analysis by changing the key model parameter values was carried out. The model simulations showed that the effective distance between the areas of hydrolysis/acidogenesis and methanogenesis is very important. An initial spatial separation of rich waste and inoculum enhances the methane production and waste degradation at high waste loading if relatively low VFA diffusion into the methanogenic area is taking place. When both hydrolysis and methanogenesis are strongly inhibited by high levels of VFA, fluctuations in biomass concentration are thought to be responsible for initiating the expansion of methanogenic area over the reactor space. PMID- 14531434 TI - Cellulosic waste degradation by rumen-enhanced anaerobic digestion. AB - Anaerobic digestion of lignocellulosic material is carried out effectively in many natural microbial ecosystems including the rumen. A rumen-enhanced anaerobic sequencing batch reactor was used to investigate cellulose degradation to give analysis of overall process stoichiometry and rates of hydrolysis. The reactor achieved VFA production rates of 207-236 mg COD/L/h at a loading rate of 10 g/L/d. Overloading of the reactor resulted in elevated production of propionic acid, and on occasion, the presence of succinic acid. With improvements in mixing and solids wasting, the anaerobic sequencing batch reactor system could enable full-scale application of the process for treatment of cellulosic waste material. PMID- 14531435 TI - Membrane separation to improve degradation of road side grass by rumen enhanced solid incubation. AB - Membrane separation proved to be an excellent means to maintain high residence time of microorganisms in an anaerobic hydrolysis reactor, and relatively low concentration of hydrolysis products. The microbial biocommunity typical for the rumen environment could be maintained, and the reactor efficiency of the reactor improved. Less than 4 days were reqired to reach almost complete hydrolysis of the grass fed into the reactor. To avoid blocking of the membrane unit, a backwash system is necessary. The membranes needed to be backwashed every 20 min with 4 bar gas-pressure for 10 s. After this treatment the initial permeability was regained. The plant was operated with a flux of 12 ml h(-1) cm(-2) on average. The transmembrane pressure was in the range of 0.8-0.9 bar. 90% of the dissolved fatty acids permeated through the membrane. PMID- 14531436 TI - Semi-continuous anaerobic digestion of food waste using a hybrid anaerobic solid liquid bioreactor. AB - A hybrid anaerobic solid-liquid (HASL) bioreactor was developed to enhance food waste conversion. The HASL bioreactor is a modified two-phase anaerobic digestion system with a solid waste reactor and a high-rate anaerobic wastewater reactor, which was a UASB reactor. In this study, the methanogenesis in the UASB reactor was investigated under a semi-continuous operation of the food waste digestion system. The HASL process, including 7 days of start-up and 23 days of semi continuous operation followed by 6 days of batch operation, was commenced by loading 2.8 kg of shredded food waste. During the semi-continuous operation, 0.2 kg of the same waste was loaded daily. The leachate from acidification phase, i.e., solid waste reactor, remained acidic (pH 4.9-5.5) and high in total VFA (TVFA), 9,500-11,500 mg/L, and COD (8,000-11,800 mg/L) levels. In the methanogenesis phase, i.e., UASB reactor, effective TVFA and COD removals (88 +/- 5% and 85 +/- 7%, respectively) were obtained, while the methane content was 71%. At the end of operation, about 78% of VS added in the HASL bioreactor was removed, while TOC and total COD reductions were 78% and 79%, respectively. The results showed that the use of UASB reactor in the semi-continuous HASL system can enhance the methanogenesis process and increase the methane content in biogas production. PMID- 14531437 TI - Phthalic acid esters found in municipal organic waste: enhanced anaerobic degradation under hyper-thermophilic conditions. AB - Contamination of the organic fraction of municipal solid waste (OFMSW) with xenobiotic compounds and their fate during anaerobic digestion was investigated. The phthalic acid ester di-(2-ethylhexyl)phthalate (DEHP) was identified as the main contaminant in OFMSW in concentrations more than half of the threshold value for the use as fertilizer on agricultural soil in Denmark. Analysis of DEHP in samples before and after large-scale anaerobic digesters revealed higher concentrations of DEHP per kg dry matter in the effluent than in the influent. The concentration of DEHP and DBP (dibutylphthalate) in OFMSW was monitored in the influent and effluent of anaerobic thermophilic (55 degrees C) and hyper thermophilic (68 degrees C) laboratory-scale reactor systems. In the thermophilic reactors with a hydraulic retention time (HRT) of 15 days 38-70% of DBP was removed, but no consistent removal of DEHP was observed. However, after treatment of the effluent from the thermophilic reactor in a hyper-thermophilic digester (HRT: 5 days) 34-53% of the DEHP content was removed and the DBP removal was increased to further 62-74%. Removal rates (k(h)) of DEHP and DBP were found to be 0.11-0.32 d(-1) and 0.41-0.79 d(-1), which is much higher than in previous investigations. It can be concluded that the higher removal rates are due to the higher temperature and higher initial concentrations per kg dry matter. These results suggest that the limiting factor for DEHP degradation is the bioavailability, which is enhanced at higher temperature and higher degradation of solid organic matter, to which the highly hydrophobic DEHP is adsorbed. The investigated reactor configuration with a thermophilic and a hyper-thermophilic treatment is, therefore, a good option for combining high rate degradation of organic matter with high biogas yields and efficient reduction of the phthalic acid ester contamination. PMID- 14531438 TI - Isolation and characterization of solventogenic, cellulase-free xylanolytic Clostridia from cow rumen. AB - Hemicellulolytic obligate anaerobes capable of converting a range of agricultural substrates and monomeric sugars of hemicellulose to solvents and acids were isolated from cow rumen fluid. The strains were identified as 5 strains of C. beijerinckii (3 different genotypes), 3 strains of C. acetobutylicum (2 different genotypes), 1 strain of C. butyricum and 3 strains of C. bifermentans (2 different genotypes). All of them produced cellulase-free xylanase activity. C. acetobutylicum LU1 produced solvent, a high of 5.1 gl(-1) and C.bifermentans LU1 produced xylanase activity a high of 4.2 IU ml(-1). PMID- 14531439 TI - Concentration of pig slurry by evaporation: anaerobic digestion as the key process. AB - Nutrient redistribution between areas with a structural pig slurry surplus and those with a shortage, is limited by the high cost of transportation and spreading, due to the high water content in slurry (more than 90%) and its relative low nutrient concentration. Water can be removed from slurry by evaporation, through the application of waste heat from a power plant or from other processes. Apart from obtaining a concentrate with an obviously higher nutrient concentration than the original slurry, another objective is to obtain clean water as condensate. The objective of this work was to study the batch vacuum evaporation of pig slurry liquid fraction, to evaluate the economic feasibility and to evaluate condensate quality as a function of both pH (4, 5 and 6) and pig slurry type (fresh slurry and anaerobically digested slurry). Results showed that condensate characteristics (ammonia nitrogen, VFA, COD) were strongly dependent on these variables. Previous anaerobic digestion presented clear advantages: it provided a fraction of the required energy and it removed organic matter, preventing its volatilisation in the evaporation process and providing higher quality condensates. These advantages make the combined treatment strategy economically more feasible than the evaporation process alone. PMID- 14531440 TI - Organic material solubilization of domestic primary sludge in anaerobic digestion at controlled pH. AB - The role of the pH in the anaerobic digestion of domestic primary sludge at mesophilic temperature was evaluated. Since solubilization of organic materials is seriously affected by pH, much research has been performed for the evaluation of significant pH influence on the production of soluble organic carbon in the acid phase. In this study, the performance of continuously stirred anaerobic reactors for the production of soluble chemical oxygen demand (SCOD) and volatile fatty acids (VFAs) in domestic primary sludge was investigated. In the first anaerobic reactor, the pH was not controlled (pH-uncontrolled) whereas the pH was fixed at 6.5 (pH-controlled) in the second reactor. Two sets were performed. Results of both sets indicated that, VSS was removed with a corresponding production of VFAs and SCOD in both reactors. However, production of VFAs and SCOD stopped earlier in the pH-controlled reactors. Methanogenic phase started to operate at earlier times whereas hydrolysis and fermentation were still operating in the pH-uncontrolled reactors. The process stability and efficiency of anaerobic digestion of substrates such as domestic primary sludge can be improved by anaerobic digestion elutriated phased treatment (ADEPT) in which the acid elutriation reactor and methanogenic reactor are separated. In this respect, ADEPT was introduced. PMID- 14531441 TI - Processing and fractional utilization of lignocellulosic substrates by "pure" and "natural and defined mixed" culture C.TM1, C.SA IV and rumen mixed culture consortia CD. AB - A detailed analysis was made of chemical fractions of common agro-residues before and after pretreatment (alkali and hydrogen peroxide), and the selective utilization of components such as WSS, EBS, TSS, lignin, cellulose and hemicellulose by pure and mixed cultures of cellulolytic and xylanolytic Clostridia was monitored and correlated with the organisms' enzyme activity. For all cultures pretreatment gave higher utilization of hemicellulose and cellulose fractions; hydrogen peroxide pretreatment was more effective than NaOH treatment. Lignin utilization was not very significant even on pretreatment. C.TM1 and C.SA IV utilized hemicellulose and cellulose better than mixed cultures in selected substrates. These results help to determine the substrate composition, pretreatment conditions and enzyme system of the organism needed when designing an inoculum for agricultural waste treatment processes such as composting or biogas generation. PMID- 14531442 TI - Chemical characteristics and methane potentials of source-separated and pre treated organic municipal solid waste. AB - A research project has investigated the biogas potential of pre-screened source separated organic waste. Wastes from five Danish cities have been pre-treated by three methods: screw press; disc screen; and shredder and magnet. This paper outlines the sampling procedure used, the chemical composition of the wastes and the estimated methane potentials. PMID- 14531443 TI - Microbiological removal of hydrogen sulfide from biogas by means of a separate biofilter system: experience with technical operation. AB - The "BIO-Sulfex" biofilter of ATZ-EVUS removes hydrogen sulfide from biogas in a biological way. Hydrogen sulfide causes massive problems during power generation from biogas in a power plant, e.g. corrosion of engines and heat exchangers, and thus causes frequent and therefore expensive engine oil changes. The BIO-Sulfex module is placed between the digester and the power-plant and warrants a cost effective, reliable and fully biological desulfurization. In the cleaned gas concentrations of less than 100 ppm can be achieved. Power-plant manufacturers usually demand less than 500 or less than 200 ppm. At present, several plants with biogas flow rates between 20 and 350 m3/h are in operation. PMID- 14531444 TI - Degradation of plant wastes by anaerobic process using rumen bacteria. AB - An operational reactor has been designed for the fermentation of a pure culture of Fibrobacter succinogenes with the constraints of strict anaerobic condition. The process is controlled by measurements of pH, redox, temperature and CO2 pressure; it allows an efficient degradation (67%) of lignocellulosic wastes such as a mixture of wheat straw, soya bean cake and green cabbage. PMID- 14531445 TI - Anaerobic digestion of solid animal waste in an accumulation system at mesophilic and thermophilic conditions, start up. AB - The anaerobic digestion of solid animal wastes has been studied in an accumulation system (AC) at a filling time of 60 days followed by about 50 days batch digestion at 40 and 50degrees C. Poor mixing conditions during anaerobic digestion of solid wastes promote stratification of the substrate and intermediate products along the reactor height. The effect of layers stratification has also been followed in the AC system. The results showed a pronounced stratification of both COD(dis) and VFA concentrations along the AC system height. The temperature had a minor effect on the methane yield. The results also showed that methanogenesis was rate limiting in the AC system while the hydrolysis was the rate-limiting step during batch digestion. PMID- 14531446 TI - Experimental investigation and modelling of the effect of sulfate on anaerobic biodegradation processes in municipal solid waste. AB - The experimental investigations have been carried out in two parts. First, the biodegradation of the organic compounds in the municipal solid waste has been investigated with focus on different sulfate concentrations influencing the methane formation. Second, the inhibition effect of hydrogen sulfide in solution (H2S(aq)) on the acetate, and methane formation, respectively, has been studied at different pH and temperature values. In solution the equilibrium of hydrogen sulfide (H2S(aq)) and the hydrogen sulfide anion (HS-(aq)) mainly depends on pH. At pH 6.3 the favoured species is H2S(aq) whereas at pH 7.7 the HS-(aq) species is favoured. Additionally, the experiments have been carried out at two different temperatures (35 degrees C and 55 degrees C). According to the acetate and methane formation the H2S(aq) species is observed to have an inhibitory effect. All simulations based on a biodegradation model represented are in good agreement with the experimental data obtained. PMID- 14531447 TI - Continuous biogas production from fodder beet silage as sole substrate. AB - Since April 2000 a two-step anaerobic plant with two subsequent 500 m3 reactors has been producing biogas from fodder beet silage (pH 3.4-4.1) as the sole substrate. The plant is located at Kirchlengern near Bielefeld, Germany. Initially the reactors were inoculated with swine manure at 37 degrees C. After a start-up phase the process was sustained at pH 7.5-8.0 by feeding the silage as sole substrate with an HRT of about 55 d twice a day. Parallel to the biogas plant at Kirchlengern four one-step laboratory reactors were continuously driven at temperatures of 37 degrees C, 45 degrees C, 60 degrees C and 65 degrees C. They were fed with the same silage, but only once per day (one impulse). The organic loading rate (OLR) was adjusted to 3.9 g volatile solids (VS)/(l*d) with a concomitant hydraulic retention time (HRT) of 27 d. There was no problem with starting the reactors, but after 86 days the volumetric gas production of the 65 degrees C reactor ceased and a high amount of approximately 65 mM propionate could be determined. By decreasing the temperature down to 60 degrees C a stable reactor performance was recovered for a period of at least 600 further days. Interestingly microscopic analyses revealed that the morphology of methanogenic bacteria in the 60 degrees C was quite different from the 37 and 45 degrees C reactor exhibiting only rodlike methanogens whereas at 37 degrees C coccoid morphotypes besides rodlike methanogens were dominant. In a 55 degrees C reactor (separate experiment) a mixture of coccoid and rodlike methanogens established. During impulse feeding with 3.9 g (VS)/(l*d) it was observed that the quickest recovery of gas production, the pH, CH4 and CO2 content as well as the redox value could be observed at 37 degrees C or at 45 degrees C. Recovery of 75% gas volume (related to the value before or after impulse feeding) was obtained after 5.5 and 7.5 h of feeding time point whereas the 60 degrees C reactor needed 16 h. Slight significant differences were seen in the spectrum of volatile fatty acids (VFA) reaching at 37 degrees or 45 degrees C its maximum with 10-30 mM total VFA at 2-3 h after feeding. After this the VFA level declined to nearly zero (except for the 60 degrees C reactor). Therefore the 37 degrees C reactor was favoured. A double experiment with a second 37 degrees C reactor was started by a somewhat different inoculation procedure from the remaining 3 reactors, but revealed similar results. An impulse feeding experiment with a very high OLR of 16.5 g (VS)/(l*d) lasting 1 week offered a stable reactor performance with a peak GPR of up to 24 l/(l reactor *d) and an HRT of 5.45d. Therefore a long term operation with an HRT of only 7.5 days and an OLR of 12 g (VS)/(l*d) should be possible. By increasing the temperature no significantly different specific gas production rates and methane yields could be observed, e.g. it gave 600-7001 biogas from 1 kg VS. The corresponding methane content ranged between 62-64%. With a methane content of 63 +/- 1% a yield of 40.1 +/- 2 m3 methane/ton fresh fodder beet silage was obtained. PMID- 14531448 TI - Methane production from sweet sorghum residues via a two-stage process. AB - The start-up of a two-stage reactor configuration for the anaerobic digestion of sweet sorghum residues was evaluated. The sweet sorghum residues were a waste stream originating from the alcoholic fermentation of sweet sorghum and the subsequent distillation step. This waste stream contained high concentration of solid matter (9% TS) and thus could be characterized as a semi-solid, not easily biodegradable wastewater with high COD (115 g/l). The application of the proposed two-stage configuration (consisting of one thermophilic hydrolyser and one mesophilic methaniser) achieved a methane production of 16 l/l wastewater under a hydraulic retention time of 19 d. PMID- 14531449 TI - Start-up of a multi-stage system for biogas production and solid waste treatment in low-tech countries. AB - Vegetable fruit garden wastes were treated anaerobically using a multistage Dranco system. The digesters were composed of three 50 L vessels kept in mesophilic conditions. They were operating at 14.5-17% TS. By controlling the pH in the system, the start-up for biogas production was shortened to 60 days. The pH correction was a buffering which enhanced methanogenic activity in the digesters. With a loading rate of 4.1 kg VS/m3 reactor/day, the production of biogas was 5 m3/m3 reactor/ day, and 60-70% methane content. This allowed making a multisystem by starting every 3 weeks with new vessels in order to maintain biogas production, to be used in industries or in local communities in low-tech countries. The designed model was started in Kinshasa (Congo) where a project is expected to treat one ton of solid waste on a daily basis, for a production of 100 m3 biogas. This cost effectiveness of the system is demonstrated and presents the opportunity for biowaste treatment coupled with environmental protection and substantial energy recovery. PMID- 14531450 TI - Effect of intermediate soil cover on municipal solid waste decomposition. AB - A complex series of chemical and microbiological reactions is initiated with the burial of refuse in a sanitary landfill. At the end of each labour day, the municipal solid wastes (MSW) are covered with native soil (or an alternative material). To investigate interaction between the intermediate cover and the MSW, five sets of columns were set up, one packed with refuse only, and four with a soil-refuse mixture (a clay loam, an organic-rich peaty soil, a well limed sandy soil and a chalky soil). The anaerobic degradation over 6 months was followed in terms of leachate volatile fatty acids, chemical oxygen demand, pH and ammoniacal N performance. Results suggest that the organic-rich peaty soil may accelerate the end of the acidogenic phase. Clay appeared not to have a significant effect on the anaerobic degradation process. PMID- 14531451 TI - Start-up of full-scale anaerobic digesters treating municipal solid waste. AB - Raising organic loading rate, and the behavior of dissolved CODcr (D-CODcr), VFA and specific methanogen activity, were investigated through a laboratory experiment for the start-up of a sludge recycling center. Moreover, application for MPN-PCR methods using a gene as a direct technique to measure the quantity of methanogen was attempted. It was recognized that specific methanogen activity depends on the quantity of methanogen, and that gas production does not reflect the condition of methane fermentation. The methane fermentation condition was checked through the specific methanogen activity and analysis of D-CODcr. The target organic loading rate was reached in the short period of about 30 days, and rapid start-up was successfully attained for a full-scale anaerobic digester. PMID- 14531452 TI - Synthesis and study of pigment aggregation response of some melatonin derivatives. AB - This study was performed to investigate the reactivity of melatonin towards various chemical reagents to produce new derivatives of melatonin containing extra- or fused heterocyclic systems such as the imidazoindole derivatives 3, 5, 6, the triazinoindole derivatives 8a, b, the thiadiazoloindole derivative 12, the pyridinoindole derivative 16 and the aminopyrazolomelatonin derivatives 21a, b and 22a, b. The structures of the compounds were established based on the analytical and spectral data. Agonist and antagonist potency of some melatonin derivatives in clonal Xenopus laevis melanophore cells was studied in comparison with melatonin. PMID- 14531453 TI - HPLC determination of naproxen in plasma. AB - An assay method using isocratic HPLC with fluorometric detection for the determination of naproxen sodium in plasma is presented. A reverse phase Microbondapack column was used with a mobile phase consisting of 42% acetonitrile and 58% water adjusted to pH 3 using phosphoric acid. The fluorometric detector with an excitation wavelength of 270 nm and emission wavelength of 340 nm provided high sensitivity and no interferences from plasma constituents. Plasma samples were injected to HPLC without any extraction. The method was precise and reproducible as was demonstrated by replicate analysis of pooled plasma sample containing 0.5-80 microg/ml naproxen sodium. PMID- 14531454 TI - HPLC analyses and pharmacokinetic studies of baicalin and oxymatrine in rabbits. AB - In the present study two simple RP-HPLC methods were developed to determine baicalin and oxymatrine in rabbit serum. Separation was performed on a Diamonsil C18 column (200 mm x 4.6 mm I.D., 5 microm) with UV detector at 277 nm for baicalin and 220 nm for oxymatrine. The mobile phase was methanol-water phosphoric acid 50:50:0.2 v/v for baicalin and acetonitrile-water (20:80, v/v, 5 mmol/L sodium octanesulfonate was contained and pH was adjusted to 3.2 with phosphoric acid for oxymatrine. p-Nitrobenzoic acid and phenacetin were used as internal standards for baicalin and oxymatrine, respectively. The standard curves were linear from 0.5 to 200.0 mg/L for baicalin and from 0.5 to 100.0 mg/L for oxymatrine with correlation coefficients of 0.9994 and 0.9965, respectively. The intra-day and inter-day RSD were less than 5.4% and 7.2% for baicalin and 6.6% and 13.8% for oxymatrine. The mean recoveries were 100.1% for baicalin and 99.1% for oxymatrine. The methods were applied to a pharmacokinetic study of baicalin and oxymatrine in rabbits. The pharmacokinetic parameters were determined after intravenous injections of baicalin and oxymatrine (40 mg/kg) separately and together to rabbits. They all fit to the two-compartment open model. Student's t test shows that there is no significant difference in the main pharmacokinetic parameters including AUC(0-infinity), when alpha and beta, when baicalin and oxymatrine were administered separately or together. PMID- 14531455 TI - A validated HPLC assay for simultaneous analysis of salmon calcitonin and duck ovomucoid. AB - A highly sensitive and selective analytical HPLC method is reported for the simultaneous measurement of salmon calcitonin (sCT) and its enzyme inhibitor, duck ovomucoid (dOVM). The method used a reversed phase C-18 column (4.6 x 250 mm, 5 microm) at room temperature. The elution was achieved using a gradient technique (20-35% B for 10 min, 35-37% B from 10th to 20th min and 37-20% B from 20th to 25th min). The mobile phase used was 0.05% v/v trifluoroacetic acid (TFA) in water and 0.05% v/v TFA in acetonitrile with a flow rate of 1 ml/min. Detection was carried out by UV spectrophotometry at 210 nm. sCT and dOVM were eluted at 7.8 and 15.4 min respectively, free from any interfering endogenous peaks during a run time of 25 min. Linear relationships were observed between the detector response and the concentrations of the analytes (10-100 microg/ml for CT (r2 = 0.996) and 10-100 microg/ml for the dOVM (r2 = 0.999)). The assay was found to be highly selective and sensitive due to the absence of any interfering peaks. The lower C.V. and % error values of the assay indicates that the assay could accurately and precisely quantitate both sCT and dOVM in the examined concentration range. This method can be usedfor the simultaneous quantitative analysis of sCT and dOVM. PMID- 14531456 TI - Simultaneous resolution of a binary mixture of captopril and hydrochlorothiazide in tablets by bivariate and multivariate spectral calibrations. AB - The multivariate spectral calibration methods, two-linear regression-calibration (bivariate calibration (BC)) and multi-linear regression-calibration (MLRC) are proposed for the simultaneous resolution of a binary mixture of hydrochlorothiazide (HCT) and captopril (CTP), which have closely overlapping spectra. The BC and MLRC calibration algorithms are described for the two component system, HCT-CTP. Some alternative methods, classical least squares (CLS), inverse least squares (ILS), principal component regression (PCR) and principal least squares (PLS) methods, were also used to determine HCT and CTP in the mixture. Using a synthetic mixture of the two drugs, all the methods were validated and applied to tablets. The BC and MLRC methods which are very rapid, and easy to apply, yet not expensive, are powerful tools with very simple mathematical contents for the quantitative analysis. Data treatment was performed using MAPLE V, EXCEL and SPSS 10.0 Software. PMID- 14531457 TI - Isolation of flavonoids from Aleurites moluccana using chitosan modified with benzaldehyde (CH-Bz) as chromatographic support. AB - This paper describes the preparation, characterization and use of a derivative of chitosan as a chromatographic sorbent. Chitosan modified with benzenic ring (CH Bz) was used to separate two flavonoids, swertisin and 2"-O-rhamnosylswertisin, from ethyl acetate fraction of Aleurites moluccana. The results showed that CH-Bz can be used as a sorbent for the separation of flavonoid compounds. The studies showed that CH-Bz in column chromatography produces goods results, separation of the flavonoid compounds. PMID- 14531458 TI - Kinetics of degradation of diclofenac sodium in aqueous solution determined by a calorimetric method. AB - An isothermal heat conduction microcalorimeter has been used to study the stability of diclofenac sodium both alone and its inclusion complex with beta cyclodextrin in aqueous solution. The rates of heat evolved during degradation of diclofenac sodium have been measured by a highly sensitive microcalorimetric technique as function of concentration, pH and temperature. The calorimetric accessible data have been incorporated in the equations for determination of rate constants, change in enthalpy and order of reaction. The decomposition of diclofenac sodium both alone and its inclusion complex with beta-cyclodextrin in solution corresponds to a pseudo-first order reaction. The values of rate constants, k's at 338.15 K, (calculated from the variation of heat evolution with the time) for the degradation of diclofenac sodium at pH 5, 6, 7, 8 and its inclusion complex with beta-cyclodextrin at pH 7 are found to be 4.71 x 10(-4), 5.69 x 10(-4), 6.12 x 10(-)4, 6.57 x 10(-4) and 4.26 x 10(-4) h(-1) respectively. There is good agreement between calorimetric determined t(0.5) and literature values. It has been found that beta-cyclodextrin retards the degradation of diclofenac sodium. The kinetic parameters have been calculated for the reaction. The negative entropy of activation suggests the formation of an ordered transition state. PMID- 14531459 TI - In vitro release of valerenic and hydroxyvalerenic acids from valerian tablets. AB - Although most commercial valerian formulations are coated tablets not any comparison study of their drug release profiles has been published so far. The main objective of this work is to establish a drug release test suitable for studying and comparing different valerian tablets. Thus, hydroxyvalerenic and valerenic acid concentrations were assayed by HPLC using a C18 Kromasil (200 x 4.6 mm, 5 microm) column and a mobile phase containing methanol and an orthophosphoric acid solution 0.5% v/v in water at a ratio of 75:25 at a constant flow rate of 1 ml/min. Saturation solubilities for hydroxyvalerenic and valerenic acid at pH 6.8 were 26 +/- 5.1 and 1 +/- 0.6 microg/ml, respectively. Usually for drugs with such low solubility values, their oral absorption and hence bioavailability are limited by their dissolution characteristics. A dissolution test was conducted according to the general method 2 (paddles) of USP 24 using 500 ml buffer medium (pH 6.8) at 50 rpm. Five different formulations were studied and compared: one uncoated tablet formulation and four marketed coated tablets. The uncoated tablet formulation had the fastest release profile, whereas the coated tablets manifested very different release patterns, depending on the type of formulation. Because of these differences in drug release pattern not every tablet formulation may be appropriate for the same clinical indications. Clinical data are required to confirm the correlation between drug release pattern and the therapeutically value of each formulation. PMID- 14531460 TI - Rofecoxib-beta-cyclodextrin inclusion complex for solubility enhancement. AB - Complex formation of rofecoxib and beta-cyclodextrin in aqueous solution and in solid state and the possibility of improving the solubility and dissolution rate of rofecoxib via complexation with cyclodextrin were investigated. Phase solubility studies indicated the formation of an 1:1 complex in solution and the value of apparent stability constant was 769 M(-1). Solid inclusion complexes of rofecoxib and cyclodextrin were prepared by the kneading method in different molar ratios. Differential scanning calorimetry studies indicated the formation of solid inclusion complexes of rofecoxib and cyclodextrin at different molar ratios and the solid complexes exhibited a higher rate of dissolution than the physical mixture and the pure drug. PMID- 14531461 TI - Modified release bi-layered tablet of melatonin using beta-cyclodextrin. AB - A modified release bi-layered tablet of melatonin incorporating a fast release fraction consisting of melatonin-beta-cyclodextrin inclusion complex and a slow release fraction containing melatonin in HPMC K15M and Carbopol 971 P matrices was prepared. The formulation developed showed an initial burst followed by a near zero order release pattern for a period of 8 h. The drug content, physical characteristics and the release profile were unaffected after 3 months of an accelerated stability study at 40 degrees C and 75% relative humidity. PMID- 14531462 TI - Particular features of photolabile substances in tablets. AB - Nifedipine and molsidomine tablets are extremely photolabile drug preparations, even at cool room light. Compared to solutions the light spectrum responsible for photodegradation is moved towards the long-wavelength range corresponding to the bathochromic shift of light absorption in the solid state. In the case of nifedipine tablets light up to 500 nm, especially the range between 400-420 nm, is degrading. Molsidomine tablets are affected only by ultraviolet light, but not by visible light. In both cases light penetrates less than 1 mm into the tablets. For nifedipine tablets the exact penetration depth could be determined due to the discolouration of the drug substance upon irradiation. It varied from 360 microm to 880 microm depending on the drug content. Since the decomposition products of nifedipine act as photostabilizers by spectral overlay, light penetration and photodegradation in nifedipine tablets are limited. The formation of gaseous and liquid decomposition products in molsidomine tablets enhances photodegradation. Changes of the tablet structure as well as dissolution and migration processes are discussed. Furthermore the degradation products donot photostabilize the drug substance due to the missing light absorption above 300 nm. PMID- 14531463 TI - Concentration dependent stereoselectivity of propafenone N-depropylation metabolism with human hepatic recombinant CYP1A2. AB - Concentration dependency of stereoselective N-depropylation metabolism of propafenone was studied by using transgenic cell line expressing human CYP1A2. Enantiomers of propafenone and N-depropylpropafenone were separated and assayed simultaneously by RP-HPLC with precolumn GITC chiral derivatization. The experimental results showed that CYP1A2 was involved in enantioselective N depropylation of propafenone and that the metabolic stereoselectivity depends on substrate concentration. For racemic propafenone, stereoselectivity was observed at low substrate concentration and was not seen at high substrate concentration. For individual isomers, S-(+)-propafenone was metabolized faster than its antipode at higher enantiomer concentrations and R-(-)-propafenone was eliminated faster than its antipode at lower enantiomer concentrations. There is interaction between S- and R-propafenone. R-(-)-propafenone inhibited the metabolism of S-(+) propafenone with IC50 0.225 mmol/L for human CYP1A2. PMID- 14531464 TI - Effect of p-amino-diphenyl ethers on hepatic microsomal cytochrome P450. AB - The present paper aims to investigate whether p-amino-2',4'-dichlorodiphenyl ether and p-amino-4'-methyldiphenyl ether are inhibitors as well as inducers of P450. Mice were given daily intraperitoneal (ip) injections of p-amino-2',4' dichlorodiphenyl ether (0.25 mmol/kg) or p-amino-4'-methyldiphenyl ether (0.25 mmol/kg) for 4 days and tested at 24 h and 48 h after the last dose injection. The results showed the mice pentobarbital sleeping time was shorter and the P450 content of hepatic microsome increased significantly in the group pretreated with p-amino-4'-methyldiphenyl ether when compared with the control group, while in mice pretreated with p-amino-2',4'-dichlorodiphenyl ether the hepatic microsome P450 content increased but the pentobarbital sleeping time was extended in clear contrast to the control group. The sleeping time of the phenobarbital group (80 mg/kg daily ip injection for 4 days) was shortened at 24 h after the last injection with increased P450 content of hepatic microsome, but it showed no difference at 48 h. The zoxazolamine-paralysis times of mice treated with p-amino 2',4'-dichlorodiphenyl ether were longer than those of the control mice, while the same dose of zoxazolamine did not lead to paralysis in mice pretreated with BNF. p-Amino-2',4'-dichlorodiphenyl ether and p-amino-4'-methyldiphenyl ether inhibited the activity of 7-ethoxyresorufin O-deethylase from rat hepatic microsome induced by BNF in vitro by 70.0% and 50.1% respectively. These results suggest that p-amino-2',4'-dichlorodiphenyl ether and p-amino-4'-methyldiphenyl ether are inhibitors as well as inducers of P450. PMID- 14531465 TI - Differences between gastric antiulcer effects of trapencaine enantiomers. AB - The spatial arrangement of single stereoisomers may influence pharmacodynamic, pharmacokinetic and toxicological properties of a drug. Trapencaine (I. N. N.), (+/-)-trans-2-(pyrrolidin-1-yl)cyclohexylester of 3-(n)-pentyloxycarbanilic acid, was developed as an antiulcer drug with gastroprotective, local anaesthetic and spasmolytic effects. Limited information is available about the potential pharmacodynamic differences of the enantiomers of trapencaine. Therefore, the enantiomers of (+/-)-trans- or cis-2-(pyrrolidin-1-yl)cyclohexylester of 3-(n) pentyloxy carbanilic acid were synthesised and tested on models of acute gastric damage induced by indomethacin and/or ethanol. A difference was found between their antiulcer effect, with the (+)-trans-enantiomer being the most effective and the (-)-cis-enantiomer the least effective in the models used. PMID- 14531466 TI - Relaxant effects of carvacrol on guinea pig tracheal chains and its possible mechanisms. AB - The bronchodilatory effects of three cumulative volumes (0.02, 0.04, and 0.08 ml) of 1/100 diluted carvacrol were examined by their relaxant effects on tracheal chains of guinea pigs precontracted by 60 mM KCl (group 1) and 10 microM methacholine in two different conditions including: non-incubated tissues (group 2) and incubated tissues with 1 microM propranolol and 1 microM chlorpheniramine (group 3), (for each group, n = 5). The relaxant effects of all three volumes of carvacrol were significantly higher than those of ethanol and even theophylline in all three groups of experiments (p < 0.05 to p < 0.001). There were positive correlations between the relaxant effect of carvacrol and incremental volume (p < 0.05 to p < 0.001). These results indicated that carvacrol has a potent relaxant effect on tracheal chains of guinea pigs which is not due to beta2-adrenergic stimulatory, histamine H1, and muscarinic blocking effect. PMID- 14531467 TI - Fast repair of the poly G hydroxyl radical adduct by two phenylpropanoid glycosides. AB - The repair activities and the reaction mechanisms of two phenylpropanoid glycosides (PPGs), isolated from a herb used in Chinese folk medicine, towards the oxidizing hydroxyl radical adduct of poly G were studied with a pulse radiolytic technique. On pulse irradiation of a nitrous oxide saturated 4 mM aqueous solution of poly G containing one of the tested compounds, the transient absorption spectrum of the hydroxyl radical adduct of poly G decays with the formation of that of the phenoxyl radical of the tested compound within several tens microseconds after the electron pulse irradiation. The results indicated that there was a repair reaction between the hydroxyl radical adduct of poly G and the compounds tested. The rate constants for the repair reactions of poly G hydroxyl radical adducts by verbasicoside and cistanoside C were determined to be 1.4 and 1.0 x 10(9) M(-1) x s(-1), respectively. PMID- 14531468 TI - 2'-(2,3-Dihydroxybenzoyloxy)-7-ketologanin: a novel iridoid glucoside from the leaves of Gentiana kurroo. AB - A new bitter acylated iridoid glucoside, 2'-(2,3-dihydroxybenzoyloxy)-7 ketologanin (1), has been isolated from the leaves of Gentiana kurroo. The structure of the compound was elucidated conclusively by chemical analysis, and extensive 1D and 2D NMR experiments. PMID- 14531469 TI - New oleanene and fernane-type triterpenes from the stem bark of Betula pendula roth. AB - A new oleanene-type triterpene, named betuloleanolic acid acetate and fernane type triterpenes, namely betufernanediol A and betufernanediol B (isomers) have been isolated from the stem bark of Betula pendula. Their structures have been established as olean-12-en-3beta-ylacetate-29-oic acid, fern-22(29)-en 3beta,12beta-diol and fem-22(29)-en-3alpha,12beta-diol, respectively, on the basis of spectral data analysis and chemical reactions. PMID- 14531470 TI - An unique n-propyl sesquiterpene from Eryngium creticum L. (Apiaceae). AB - The unique 1-n-propyl-perhydronaphthaline 1,2,4a,5,6,7,8,8a-octahydro-4-methyl-1 propyl-naphthalene-7-carbaldehyde, a compound which possesses an unusual sesquiterpene carbon skeleton, was isolated and identified, together with the new natural methyl ketone eicos-8,11 -dien-18-ol-2-one, from the hexane: ether extract of the aerial parts of Eryngium creticum growing in Sinai, Egypt. The structures were established by conventional methods of analysis and confirmed by DEPT, COSY, HMQC and HMBC. PMID- 14531471 TI - Effect of enhancers on percutaneous absorption of osthol across the excised full thickness rat skin. PMID- 14531472 TI - Chemcial constituents from the marine sponge Iotrochoto birotulata. PMID- 14531473 TI - Flavonoids of Pueraria lobata: chromatographic analysis of leaves and roots of cultivated plants. PMID- 14531474 TI - Activation of human polymorphonuclear neutrophils by environmental contaminants. AB - The toxicity of chemicals of environmental concern to the immune system has been primarily evaluated in animals and, to a lesser extent, in humans. In particular, the effects of various pollutants on B-cell, T-cell, natural killer cells, and monocyte-macrophage cells have been the focus of several reports, but polymorphonuclear neutrophils have largely been neglected. Recent data indicate that neutrophils are important targets for such chemicals, suggesting a potential role of these products in the development of the inflammatory process. The bulk of this review will focus on the role of certain environmental pollutants on human neutrophil cell physiology. PMID- 14531475 TI - The role of combined metal interactions in metal carcinogenesis: a review. AB - Exposures to complex mixtures of metals in the workplace or environment are more likely to occur than exposures to a single metal alone. The evidence shows that exposures to complex metal mixtures can enhance the risk of cancer in certain human populations. The findings of several studies have suggested, however, that certain metal-metal interactions can inhibit carcinogenic activity. The mechanisms of metal-metal interactions in human carcinogenesis are relatively unknown. Metals represent a highly diverse group of agents: each metal can act through different mechanisms and in one or more steps of the carcinogenic process. Some potential mechanisms may involve direct reactions of the metal with DNA or indirect mechanisms that include modification of DNA repair, DNA methylation status, and metabolic processes involved in DNA replication and expression. Lipid peroxidation and the generation of free radicals induced by certain metals can affect DNA integrity. This review will address the role of metals in carcinogenesis and how concomitant exposure to metal mixtures can influence cancer induction. The most current mechanistic data regarding metal interactions and its implications in human carcinogenesis will be discussed. Furthermore, research gaps will be identified to provide data that will improve risk assessments for complex metal mixtures encountered in the workplace and environment. PMID- 14531476 TI - Inadequate levels of essential nutrients in developed nations as a risk factor for disease: a review. AB - Inadequate levels of essential nutrients is a most important factor in environmental health, leading to an almost monotonic increase in the incidence, morbidity, mortality, and associated costs of 'diseases of affluence' that has persisted for circa a century. Such an unprecedented human tragedy can be explained by the (1) flawed belief that essential nutrients can be replaced by xenobiotic molecules--a fundamental error, (2) failure to recognize that the levels of certain essential nutrients in diets available in the environment are not adequate to produce optimum health, longevity or resistance, and (3) failure to recognize that the refined diet has a lower essential nutrient content. Such beliefs contribute to early death in affluent nations. PMID- 14531477 TI - Molecular microbial biodiversity assessment: a biological indicator of soil health. AB - The soil performs a variety of key functions: (i) provides the food, fuel, and fiber needs of the world's population, (ii) regulates the quality of the air and water, (iii) decomposes organic wastes, (iv) recycles nutrients, and (v) acts as a sink for pollutants (including global gases). Soil degradation is increasingly recognized as an urgent environmental issue and a crucial need exists for the capacity to evaluate soil health. After all, soil health is a key component in sustaining the different world's ecosystems and the myriad of natural and socioeconomic systems they support. In this respect, soil health has been defined as the "continued capacity of a specific kind of soil to function as a vital living system, within natural or managed ecosystem boundaries, to sustain plant and animal productivity, to maintain or enhance the quality of air and water environments, and to support human health and habitation". A biological indicator is an organism, a part of an organism, or a community of organisms, used to obtain information about environmental quality. The assessment of microbial biodiversity has the potential to provide useful insight into the health and functioning of soil. Our inability to culture most microorganisms that are present in soils has, until recently, impaired studying the relationships between the structure and the function of soil microbial communities. This shortcoming has been recently overcome by using several molecular techniques that allow the detection, enumeration, and characterization of soil microorganisms without cultivation. PMID- 14531478 TI - Fever and asymmetrical weakness in the summer: evidence of a West Nile virus associated poliomyelitis-like illness. PMID- 14531479 TI - Immediate and long-term hemodynamic and clinical effects of sildenafil in patients with pulmonary arterial hypertension receiving vasodilator therapy. AB - OBJECTIVE: To determine the immediate and long-term effects of adding sildenafil, a phosphodiesterase-5 inhibitor, to the medical regimen of patients with pulmonary arterial hypertension (PAH). PATIENTS AND METHODS: Thirteen patients with PAH received empirical adjunctive sildenafil treatment at the Mayo Clinic in Rochester, Minn, between November 1, 2000, and August 31, 2001. All received a 25 mg dose of sildenafil, increased by 25 mg at 8-hour intervals, if tolerated, up to 100 mg during hemodynamic monitoring for 24 to 48 hours. Long-term effects on right heart hemodynamics were assessed by noninvasive right ventricular systolic pressure, right ventricular index of myocardial performance, and a 6-minute walk test. RESULTS: Sildenafil significantly increased cardiac output (CO) (P = .04) and decreased pulmonary artery systolic pressure, mean pulmonary artery pressure, pulmonary vascular resistance, and mean arterial pressure (P < or = .01) at peak measurements (obtained 1-2 hours after highest dose). At trough measurements (obtained 8 hours after highest dose), sildenafil significantly decreased pulmonary artery systolic pressure, mean pulmonary artery pressure, and mean arterial pressure (P = .01). Ten patients discharged from the hospital were taking the highest-tolerated dose of sildenafil every 8 hours. The right ventricular systolic pressure and right index of myocardial performance showed no significant improvement at follow-up (117 +/- 70 days), although concomitant treatment with epoprostenol could be tapered in 2 patients. Changes in New York Heart Association classes were inconsistent, and improvements in the 6-minute walk test were not significant. CONCLUSION: Sildenafil has an immediate pulmonary vasodilator effect in patients already receiving vasodilators for PAH. Its long term effects on right heart function and functional status are equivocal. A large, prospective, well-designed study is needed to determine the effects of sildenafil on PAH, both in untreated and concurrently treated patients. PMID- 14531480 TI - Pain-free results with sumatriptan taken at the first sign of migraine pain: 2 randomized, double-blind, placebo-controlled studies. AB - OBJECTIVE: To evaluate the efficacy and tolerability of sumatriptan, 50-mg and 100-mg tablets, compared with placebo for treatment of migraine at the first sign of pain. PATIENTS AND METHODS: Two identical multicenter randomized, double blind, placebo-controlled, single-attack studies were conducted from May through November 2000 in adults (aged 18-65 years). Patients treated migraine at the first sign of pain, while pain was mild, but not more than 2 hours after onset with oral sumatriptan, 50 mg or 100 mg, or matching placebo. The primary end point was pain-free relief at 2 hours after treatment with 50 mg of sumatriptan compared with placebo. RESULTS: There were 354 patients in study 1 and 337 patients in study 2. Significantly more patients treated with sumatriptan, 50 mg and 100 mg, were completely free from pain 2 and 4 hours after treatment vs patients treated with placebo (at 2 hours, 50% and 57% vs 29%; at 4 hours, 61% and 68% vs 30%; for both, P < .001). Also, significantly more patients treated with sumatriptan, 50 mg and 100 mg, were migraine-free (no pain or associated symptoms) vs those treated with placebo at 2 and 4 hours after treatment (at 2 hours, 43% and 49% vs 24%; at 4 hours, 54% and 63% vs 28%; for both, P < .001). The incidence of overall adverse events was low with the 50- and 100-mg dose of sumatriptan (placebo, 7%; sumatriptan at 50 mg, 14%; sumatriptan at 100 mg, 16%). CONCLUSIONS: Treatment of migraine at the first sign of pain with sumatriptan, 50 mg and 100-mg tablets, provides superior pain-free relief at 2 and 4 hours after treatment compared with placebo. Results of these studies suggest that sumatriptan at 100 mg may be more efficacious than at 50 mg when used in the early treatment paradigm. Because these studies were not powered to detect statistical differences between active doses, studies to investigate this finding are warranted. PMID- 14531481 TI - Nonhepatosplenic extramedullary hematopoiesis: associated diseases, pathology, clinical course, and treatment. AB - OBJECTIVE: To define associated clinical conditions, pathology, natural history, and treatment outcome of nonhepatosplenic extramedullary hematopoiesis (NHS-EMH). PATIENTS AND METHODS: We retrospectively reviewed the medical charts of all patients identified as having NHS-EMH from 1975 to 2002. Diagnosis was made by tissue biopsy, fine-needle aspiration biopsy, or radionuclide bone marrow scanning. RESULTS: We identified 27 patients with antemortem diagnosis of NHS EMH. The most common associated condition and disease site were myelofibrosis with myeloid metaplasia (MMM) (in 18 patients [67%]) and the vertebral column (in 7 patients [26%]; all involving the thoracic region), respectively. At the time of diagnosis of NHS-EMH, concurrent splenic EMH (in 22 patients [82%]; 15 [56%] had undergone splenectomy) and red blood cell transfusion dependency (in 12 patients [44%]) were prevalent. Of the 27 patients, 9 (33%) required no specific therapy. Specific therapy was radiation (in 7 patients with a 71% response rate) and surgical excision (in 6 patients with a 67% response). Treatment-associated complications were limited to surgery. Radiation therapy was not used in the non MMM group, but low-dose radiation therapy was used in the MMM group for paraspinal or intraspinal EMH (median dose, 1 Gy; range, 1-10 Gy), pleural or pulmonary disease (median dose, 1.25 Gy; range, 1.00-1.50 Gy), and abdominal or pelvic disease (median dose, 2.02 Gy; range, 150-4.50 Gy). Median survival after the diagnosis of NHS-EMH was 13 months in the MMM group and 21 months in the non MMM group. CONCLUSIONS: This retrospective study suggests that NHS-EMH is rare, is often associated with MMM, and preferentially affects the thoracic spinal region. Asymptomatic disease may require no specific treatment, whereas symptomatic disease is best managed with low-dose radiation therapy. PMID- 14531482 TI - Endovascular repair of abdominal aortic aneurysms: initial experience with 100 consecutive patients. AB - OBJECTIVE: To review early results of endovascular repair of abdominal aortic aneurysms (AAAs). PATIENTS AND METHODS: The first 100 patients who underwent endovascular repair of AAA (EVAR) between June 26,1996, and October 31, 2001, at the Mayo Clinic in Rochester, Minn, were studied retrospectively to evaluate technical success, freedom from reinterventions, and early clinical outcome. RESULTS: A total of 89 men and 11 women (mean +/- SD age, 76 +/- 7 years; range, 47-92 years) underwent EVAR. The procedure was successful in 97 patients. There was no early death. Major complications occurred in 25 patients. The 30-day technical success rate was 86% (95% confidence interval [CI], 77%-92%). The median intensive care unit stay was 1 day (range, 1-15 days), and the median hospital stay was 3 days (range, 1-35 days). Median follow-up was 7 months (range, 1-60 months). Endoleak (incomplete seal of the endovascular graft) at discharge was observed in 14 patients; 13 developed endoleak during follow-up. There were 23 reinterventions, 65% of which were percutaneous procedures. One aneurysm ruptured at 5 months, but the patient was successfully treated by open repair. Primary and secondary graft patency rates at 1 year were 83% (95% CI, 74% 93%) and 94% (95% CI, 87 %-99%), respectively. The freedom from reintervention rate at 1 year was 71% (95% CI, 59%-84%), with an overall success rate from EVAR of 92% (95% CI, 84%-100%). There were no differences in early patency, reinterventions, and success rates between unibody and modular devices. CONCLUSION: EVAR can be performed with high technical success and low mortality rates; however, nonfatal complications and catheter-based reinterventions are frequent, and EVAR may not prevent aneurysm rupture. Although stent graft repair for high-risk patients is appealing, current data are insufficient to support EVAR as the preferred treatment of AAAs. PMID- 14531483 TI - Clinical and electrophysiologic patterns of flaccid paralysis due to West Nile virus. PMID- 14531484 TI - Minimally invasive procedures for disorders of the lumbar spine. AB - In the past decade, there has been a substantial increase in interest in minimally invasive procedures in all areas of medicine, particularly for spinal disorders. Some of these techniques represent notable advances in spinal care and have major roles in the care of patients with back-related symptoms. Other techniques appear to offer no benefit and in some cases may be less effective than conventional treatments. Percutaneous lumbar diskectomy techniques hold considerable promise; however, lumbar microdiskectomy is the gold standard for surgical treatment of lumbar disk protrusion with radiculopathy. Intradiskal electrothermal therapy is emerging as a useful option for selected patients with intractable mechanical back pain whose only other option historically has been a spinal fusion. Percutaneous fusion techniques are in their infancy and may prove to be beneficial for these patients as well. Percutaneous vertebral augmentation, including vertebroplasty and kyphoplasty, has become the treatment of choice for many patients with intractable back pain secondary to vertebral insufficiency fractures. Spinal injections are important for evaluating and managing spinal pain and can be extremely useful diagnostically and therapeutically. This multidisciplinary review outlines the status of these procedures and offers suggestions for their use in patient care. PMID- 14531485 TI - Dehydroepiandrosterone: is there a role for replacement? AB - Dehydroepiandrosterone (DHEA) and its sulfated ester are found in high concentrations in the plasma; however, their role in normal human physiology, other than as precursors for sex hormones, remains incompletely defined. Studies of rodent models have shown that these hormones have beneficial effects on a wide variety of conditions, such as diabetes, obesity, immune function, atherosclerosis, and many of the disorders associated with normal aging. However, rodents are not the best models to study the actions of these hormones because they have very little endogenous DHEA; thus, the doses given to these animals are usually suprapharmacological. Human studies have been performed to determine the potential beneficial effects of DHEA replacement in persons with low DHEA levels. Results have been conflicting. Human studies suggest a potential role for DHEA replacement in persons who have undergone adrenalectomy and possibly in the aging population. However, long-term studies assessing the benefits vs adverse effects must be done before DHEA replacement can be recommended. PMID- 14531486 TI - Anemia in adults: a contemporary approach to diagnosis. AB - There are numerous ways of classifying the causes of anemia, and no one way is necessarily superior to another. It is equally important to appreciate the differences in the approaches to diagnosis between children and adults, men and women, and persons of different ethnic backgrounds. Regardless of the specific algorithm followed in evaluating anemia, it is essential that easily remediable causes such as nutritional deficiencies, hemolysis, and anemia of renal insufficiency are identified early and treated appropriately. In general, the differential diagnosis of anemia can be substantially narrowed by subcategorization into "microcytic," "normocytic," and "macrocytic" subtypes on the basis of mean corpuscular volume. However, such classification is a starting point and not infallible. Each category then can be deciphered using a stepwise approach that utilizes readily accessible laboratory tests. PMID- 14531487 TI - 39-year-old man with human immunodeficiency virus infection and abdominal pain. PMID- 14531488 TI - Essentials of the proper diagnoses of mild cognitive impairment, dementia, and major subtypes of dementia. AB - Loss of cognitive function in the elderly population is a common condition encountered in general medical practice. Diagnostic criteria and approaches have become more refined and explicit in the past several years. Precise diagnosis is feasible clinically. In this article, the precursor state and major subtypes of dementia are considered. Mild cognitive impairment is the term given to patients with cognitive impairment that is detectable by clinical criteria but does not produce impairment in daily functioning. When daily functioning is impaired as a result of cognitive decline, dementia is the appropriate syndromic label. Specific causes of dementia tend to have distinctive clinical presentations: the anterograde amnesic syndrome of Alzheimer disease; the syndrome of dementia with cerebrovascular disease; the syndrome of Lewy body dementia with its distinctive constellation of extrapyramidal features, disordered arousal, and dementia; the behavioral-cognitive syndrome of frontotemporal dementia; the primary progressive aphasias; and the rapidly progressive dementias. Because dementia syndromes have distinctive natural histories, precise diagnosis leads to a better understanding of prognosis. As new treatments become available for Alzheimer disease, the most common of the dementias, accurate diagnosis allows the appropriate patients to receive treatment. PMID- 14531489 TI - Needle ingestion via Halloween caramel apples. PMID- 14531490 TI - Old age in America. PMID- 14531491 TI - Local or long distance? PMID- 14531492 TI - Chemomodulatory influence of Ferula asafoetida on mammary epithelial differentiation, hepatic drug metabolizing enzymes, antioxidant profiles and N methyl-N-nitrosourea-induced mammary carcinogenesis in rats. AB - The present study was conducted to ascertain the modulatory influences of Ferula asafoetida L. (asafoetida, flavoring agent) on the mammary epithelial tissue differentiation, hepatic drug metabolizing enzymes, antioxidant profiles and N methyl-N-nitrosourea (MNU)-induced mammary carcinogenesis in Sprague-Dawley rats. Feeding with two doses of asafoetida (1.25 and 2.5% w/w in diet) showed a remarkable increase in the development and differentiation of ducts/ductules (p < 0.01-0.005), lobules (p < 0.005) and a decrease in terminal end buds (p < 0.05 0.005) as compared to both normal and MNU-treated control animals. To assess the biochemical parameters, effect of asafoetida on drug-metabolizing enzymes was evaluated in the liver of rats. Asafoetida treatment significantly reduced (p < 0.05) the levels of cytochrome P450 and b5. There was an enhancement in the activities of glutathione S-transferase (p < 0.05-0.005), DT-diaphorase (p < 0.05 0.01), superoxide dismutase (p < 0.01-0.005) and catalase (p < 0.05-0.005) and in the level of reduced glutathione (p < 0.05-0.005), followed by asafoetida treatment. Also, asafoetida significantly restored the level of antioxidant system, depleted by MNU-treatment. The strengthening of antioxidant system by the lower and higher doses of asafoetida in the presence and absence of MNU was further substantiated by a significant inhibition (p < 0.005) in lipid peroxidation as measured by thiobarbituric acid-reactive substances (TBARS) in the liver of rat. Further, in long-term animal studies, where MNU was used to induce mammary carcinogenesis, asafoetida treatment resulted in a significant reduction in the multiplicity (p < 0.001) and size of palpable mammary tumors (p < 0.005-0.001) and a delay in mean latency period of tumor appearance (p < 0.005). Together, these findings indicate the chemopreventive potential of asafoetida against MNU-induced mammary carcinogenesis. Thus, asafoetida needs further investigation with regard to identification and characterization of its active principle(s) and mechanism of action, for this compound to be developed as a potential chemopreventive agent for human cancers. PMID- 14531493 TI - High energy shock waves (HESW) enhance paclitaxel cytotoxicity in MCF-7 cells. AB - High energy shock waves (HESW) produced by a piezoelectric generator were studied for their effect on human breast cancer cell (MCF-7) viability and sensitivity to paclitaxel. A dose-dependent impairment of cell viability was observed after HESW treatment (250-2000 shock waves, rate = 4/s, energy flux density = 0.25 mJ/mm2). Single treatment with shock waves produced no significant growth inhibition. Combined exposure to paclitaxel (ranging 0.1 nM to 20 microM) and shock waves (100, 500 and 1000 shots, respectively) resulted in a significant reduction of MCF-7 cell proliferation at day 3 after treatment in respect with cells treated with paclitaxel alone. Notably, a cell viability reduction of about 50% was obtained after combined treatment with HESW and 10 nM paclitaxel, in front of a reduction of only 40% using 10 microM paclitaxel alone. Moreover, an earlier induction as well as an enhancement of apoptotis was observed in cells subjected to combined treatment with shock waves and paclitaxel (200 nM; 20 microM). In conclusion, HESW can enhance paclitaxel cytotoxicity in MCF-7 cells, thus allowing the treatment with lower doses of drug. PMID- 14531494 TI - Racial disparity in the dose and dose intensity of breast cancer adjuvant chemotherapy. AB - PURPOSE: The purpose of this study was to investigate the impact of race and obesity on dose and dose intensity of adjuvant chemotherapy. METHODS: We abstracted data on patient/tumor characteristics, treatment course, physicians' intention to give a first cycle dose reduction, and reasons for dose reductions/delays from oncology records of 489 women treated from 1985 to 1997 in 10 treatment sites in two geographical regions. Administered doses and dose intensity were compared to standard regimens. Multivariate regression models determined the impact of race and body mass index (BMI) on dose proportion (actual:expected doses) and relative dose intensity (RDI) controlling for patient characteristics, comorbidity, chemotherapy regimen, site, and year of treatment. Logistic regressions explored race and BMI versus use of first cycle dose reductions. RESULTS: African-Americans received lower chemotherapy dose proportion and RDI than whites (0.80 vs. 0.85, p = 0.03 and 0.76 vs. 0.80, p = 0.01). In multivariate analyses, dose proportion was 0.09 lower (p = 0.002), and RDI was 0.10 (p < 0.001) lower in non-overweight African-Americans than whites. Obesity was associated with lower dose proportion (p < 0.01) and RDI (p < 0.03). Race and BMI were independently associated with first cycle dose reductions. Non overweight African-Americans (p < 0.05) and overweight and obese African-American and white women (p < 0.001) were more likely to have first cycle dose reductions than non-overweight whites. CONCLUSION: We identified systematic differences in the administration of chemotherapy given to African-Americans and to overweight and obese women. These differences may contribute to documented disparities in outcome. PMID- 14531495 TI - Role of chest X-ray in diagnosis of the first breast cancer relapse: a randomized trial. AB - Between May 1991 and December 1995, follow-up after primary therapy for 472 consecutive patients with localized breast cancer was randomly assigned to assess the efficacy of routine chest X-ray in detecting intrathoracic relapse as the first metastatic event. One group had regular chest X-rays while the other group had chest radiographs only when clinically needed (spontaneous). Patients were followed up until December 1999 or death. In the routine group, 243 patients had 1429 chest X-rays (mean 5.9 chest X-rays per patient). In the spontaneous group, 229 patients had 411 chest X-rays (mean 1.8 chest X-rays per patient). Both sensitivity and specificity were somewhat higher in the spontaneous arm compared to the routine arm. Patient sensitivity was 30% in the regular arm and 36% in the spontaneous arm. Film sensitivity was 11% in the regular arm and 20% in the spontaneous arm. Patient specificity was 85% in the regular arm and 86% in the spontaneous arm. The predictive values were practically the same in both arms. Patient positive predictive values were 22% in both arms and film positive predictive values were 21% in the regular and 22% in the spontaneous arm. Patient negative predictive values were 90% in the regular and 92% in the spontaneous arm. Film negative predictive values were 93% in the regular and 89% in the spontaneous arm. The differences in accuracy were not statistically significant between the arms. There were no significant differences in disease free survival or overall survival. The 5-year disease free survival was 86% in the regular and 89% in the spontaneous arm and the overall survival was 88 and 85% in the regular versus spontaneous arm, respectively. Routine chest X-ray in the follow-up of breast cancer patient is evidently of little use and is not likely to be of benefit to the patient in terms of disease free survival or overall survival as compared to the spontaneous schedule. PMID- 14531496 TI - A pilot study of dose intense doxorubicin and cyclophosphamide followed by infusional paclitaxel in high-risk primary breast cancer. AB - We conducted a pilot study of dose dense doxorubicin and cyclophosphamide (AC) combination chemotherapy followed by infusional paclitaxel (T) in primary breast cancer to determine its safety and feasibility. Twenty-two subjects (10 with stage II and > or = 4 positive lymph nodes, and 12 with stage III disease) were treated with AC (A 60 mg/m2 and C 2000 mg/m2) with filgrastim every 14 days for three cycles followed by infusional paclitaxel (140 mg/m2 over 96 h) every 14 days for three cycles. Mean overall cycle length was 15.3 days and mean duration of therapy was 92 days. Dose reductions of C or T were required in 7/132 (5.3%) cycles for mucositis, diarrhea, or failure to recover platelets by day 15. Ninety five percent of subjects had grade 4 neutropenia and 1 subject had a platelet nadir of < 20,000. Actual delivered dose intensity (DI) over six cycles was: A 27 mg/m2 per week; C 892 mg/m2 per week; T 64 mg/m2 per week (90.6, 89.2, and 91.4% of planned DI, respectively). Average total dose administered was: A 180 mg/m2; C 5880 mg/m2; T 403 mg/m2 (100, 98, and 96% of planned total doses, respectively). Clinical response rate in 10 subjects receiving neoadjuvant therapy was 100% (4 complete response, 6 partial response). Four subjects had a pathologic complete response (three subjects without evidence of malignancy and one subject with ductal carcinoma in situ.) Administration of dose dense AC followed by infusional paclitaxel in 14-day cycles is feasible and this regimen is active in breast cancer. PMID- 14531497 TI - Prevalence and spectrum of p53 mutations in white Hispanic and non-Hispanic women with breast cancer. AB - Hispanic women differ from non-Hispanics in breast cancer incidence, stage at diagnosis, and survival. Ethnic differences in genetic makeup, reproductive patterns, diet, socioeconomic status, physical activity, and other unidentified cultural factors may be responsible for the disparity. This study investigated occurrences of p53 tumor suppressor gene mutations in South Florida white Hispanic and white non-Hispanic women with primary breast cancer. Tumor tissues were obtained from a consecutive series of women with breast cancer who underwent breast resection at the Jackson Memorial Hospital, Miami, Florida between 1984 and 1986. A total of 231 women with primary breast cancer, aged 31-85 years, were included in the study. Among them, 64 (27.7%) were white Hispanic and 167 (72.3%) were white non-Hispanic. The majority of the patients were white non-Hispanics (72.3%). Compared to white non-Hispanics, however, white Hispanics had significantly higher proportions of tumors larger than 2 cm (53.1% v.s. 28.7%, p = 0.00) as well as larger tumor size at diagnosis (mean: 4.2 v.s. 3.0 cm, p = 0.00). The p53 gene mutation rate was significantly lower in white Hispanics than in white non-Hispanics (51.6% v.s. 70.7%, p = 0.01). Furthermore, among node negative breast cancer patients, after adjustment for tumor size at diagnosis, logistic regression results showed that white Hispanics were 71% less likely than white non-Hispanics to carry p53 mutations (OR = 0.29 and 95% CI = 0.09-0.91). We conclude that white Hispanic women with breast cancer might have lower p53 gene mutation prevalence than white non-Hispanic women. PMID- 14531498 TI - Necrosis and hypoxia in invasive breast carcinoma. AB - To investigate the relation between necrosis and hypoxia in breast cancer we examined the expression of hypoxia-associated markers HIF1, CA IX and GLUT1 by immunohistochemistry in 97 invasive ductal carcinomas. This selected series comprised 48 tumors with extensive necrosis and 49 control tumors without necrosis. Over 90% of necrotic and 30% of non-necrotic tumors expressed at least one hypoxia marker. We also observed expression of hypoxia associated markers in tumor stroma. Examination of primary human breast fibroblasts in vitro confirmed that CA IX mRNA and protein can be induced by hypoxia. Survival analysis of 53 cases found that the subset of tumors with stromal hypoxia exhibit better prognosis (p=0.027). Our results indicate that necrosis is often accompanied by hypoxia but that hypoxia without necrosis may also be a frequent occurrence. The use of several hypoxia markers may identify a continuum of hypoxia in tumors, which can be sub-classified by different co-expression patterns. We conclude that stromal and epithelial hypoxia may have different biological backgrounds and that stromal hypoxia may affect survival. PMID- 14531499 TI - Different expressivity of BRCA1 and BRCA2: analysis of 179 Italian pedigrees with identified mutation. AB - Mutations in BRCA1 and BRCA2 show different expressivity with respect to cancer risk, and allelic heterogeneity may be present in both genes. We collected 179 pedigrees with identified germline mutation (104 BRCA1 and 75 BRCA2), ascertained in six collaborating centers of the Italian Consortium for Hereditary Breast and Ovarian Cancer. Significant heterogeneity was detected for several variables, and a logistic regression model including age of diagnosis in the proband, presence of ovarian cancer in the family, presence of prostate or pancreatic cancer in the family, and presence of male breast cancer in the family proved to be effective in predicting the presence of a mutation in a gene rather than the other. Excess of familial aggregation of both breast and ovarian cancer was observed in both genes. Proportion of ovarian cancer was increased in the 5' portion of BRCA1, and presence of prostate or pancreatic cancer in a family was correlated with presence of ovarian cancer in BRCA2. PMID- 14531500 TI - Oestrogen receptor-mediated modulation of the EGFR/MAPK pathway in tamoxifen resistant MCF-7 cells. AB - Oestrogen receptor (ER) levels are usually maintained on acquisition of tamoxifen resistance in the clinic, however, tumour re-growth is associated with increased expression of epidermal growth factor receptor (EGFR) and activation of the mitogen activated protein kinase (MAPK) pathway. In the present study we have used the ER down-regulator fulvestrant ('Faslodex') to investigate the influence of the ER on growth of a tamoxifen-resistant (TAM-R) human breast cancer cell line. Expression levels of ER mRNA and protein were equivalent in parental wild type MCF-7 (WT) and TAM-R cells. Fulvestrant eliminated ER protein expression and inhibited proliferation in both cell lines. The growth inhibitory effects of fulvestrant were associated with a decrease in basal EGFR, c-erbB2 and ERK1/2 activity in TAM-R but not WT cells. ER functionality as determined by oestrogen response element (ERE)-luciferase reporter activity and expression of PgR, pS2 and transforming growth factor alpha (TGFalpha) was significantly reduced in TAM R compared to WT cells and was further decreased by fulvestrant treatment in both cell lines. Epidermal growth factor (EGF) and TGFalpha significantly increased EGFR/MAPK pathway activity in both cell lines. Ligand-induced EGFR/MAPK activation promoted TAM-R cell growth in both the absence and presence of fulvestrant, whereas no proliferative activity was observed under the same conditions in WT cells. These results suggest that the ER modulates EGFR/MAPK signalling efficiency in TAM-R cells possibly through the regulation of TGFalpha availability. This effect may be overcome by the action of exogenous EGFR ligands, which strengthen EGFR/MAPK signalling activity to generate endocrine insensitive cell growth. PMID- 14531501 TI - Research ethics--revisited. PMID- 14531502 TI - Treating patients with functional somatic symptoms. A treatment guide for use in general practice. AB - Patients who experience body symptoms in response to psychosocial stress are a challenge for the general practitioner (GP). This paper is a contribution to the improved treatment of these patients. It presents a specific psychosocial treatment model provided by the GP including the following steps: (a) taking a thorough symptom history, conducting a psychosocial anamnesis and exploring the patient's own perception of the illness, (b) developing alternative perceptions of the illness by understanding the psycho-physiological relationship; and (c) reducing the impact of psychosocial stress. The application of these specific techniques requires a trustful, helping alliance between the doctor and the patient and cooperation with mental health services. PMID- 14531504 TI - Implementation of a clinical dementia guideline. A controlled study on the effect of a multifaceted strategy. AB - OBJECTIVE: To assess the impact of a multifaceted implementation strategy aiming to improve GP adherence to a clinical guideline on dementia. DESIGN: Controlled before and after study using data records from regional laboratories. The guideline was mailed to all GPs. The multifaceted implementation strategy was planned with local GPs, and consisted of seminars, outreach visits, reminders and continuing medical education (CME) small group training. SETTING: Primary health care. SUBJECTS: 535 GP practices with 727 physicians in Denmark. MAIN OUTCOME MEASURES: The diffusion and use of the guideline was measured by a mailed survey. Adherence to guideline recommendations was monitored by data on laboratory tests from general practice in patient's > or = 65 years: thyroid stimulating hormone requested with vitamin B12 or methylmalonate. The use of these tests as part of a diagnostic evaluation of dementia was subsequently verified by a questionnaire to the practices. RESULTS: Of the GPs who read the guideline, 88% found it applicable in primary care. No increase in the adherence to guideline recommendations was observed regarding the use of laboratory tests or cognitive tests in the diagnostic evaluation of dementia in general practice. CONCLUSION: Although GPs regarded the guideline applicable in primary care, no change in practice adherence to guideline recommendations was detected after a multifaceted implementation. PMID- 14531503 TI - Anxiety and depression in individuals with somatic health problems. The Nord Trondelag Health Study (HUNT). AB - OBJECTIVE: To examine the relationship between anxiety disorders and depression and various somatic health problems in the general population. DESIGN: Cross sectional study with survey methods and clinical examinations. SETTING: The Health Study of Nord-Trondelag, Norway (the HUNT study). PARTICIPANTS: 60869 individuals aged 20-89 years. MAIN OUTCOME MEASURES: Anxiety disorder, depression and their comorbidity are categorized based on scores on the Hospital Anxiety and Depression Scale. All somatic health variables are self-reported, while blood pressure, height and weight are measured. Multivariate nominal logistic regression analyses are used to investigate the relationship between somatic variables and the anxiety/depression categories. RESULTS: Most somatic health variables show a stronger association with comorbid anxiety disorder/depression than with anxiety disorder or depression alone. About one-third of individuals reporting somatic health problems also have anxiety disorder and/or depression. CONCLUSION: Somatic health problems carry a high risk of both anxiety disorder and depression. Active identification and treatment of these co-occurring mental disorders are of practical importance. PMID- 14531505 TI - Missing the diagnosis: senile dementia in patients admitted to nursing homes. AB - OBJECTIVE: To establish dementia diagnoses in patients recently admitted to a nursing home by a geriatrician, and to compare the agreement with diagnoses recorded in the nursing home's medical records and with short screening instruments. DESIGN: Descriptive study. Geriatric work-up of the individual patient, information obtained from the medical records of nursing homes, structured interviews with nurses and a short cognitive test. SETTING: Nursing homes in Bergen, Norway, with 123 long-term care patients. MAIN OUTCOME MEASURES: Dementia diagnosis according to the International Classification of Diseases (ICD 10), Clinical Dementia Rating and the Short Portable Mental Status Questionnaire. RESULTS: Fifty-nine percent of the patients were diagnosed as demented according to the ICD-10. One-third of cases were not formerly diagnosed according to nursing home medical records. Clinical Dementia Rating and the Short Portable Mental Status Questionnaire detected "dementia/non-dementia" cases correctly in 93% and 94%, respectively. CONCLUSION: Geriatric work-up indicates under diagnosing of dementia among recently admitted nursing home patients. This may have serious consequences for the treatment. Dementia diagnostics in primary health care must be improved. The knowledge of the attending nurses should be better incorporated in the medical work-up of nursing home patients. PMID- 14531506 TI - Exposure to potential drug interactions in primary health care. AB - OBJECTIVE: To analyse the prevalence of potential drug interactions, and to identify patients particularly prone to drug interaction. DESIGN: Database study (Odense University Pharmacoepidemiologic Database). SETTING: Individuals exposed to polypharmacy in 1999 were examined for potential drug interactions. SUBJECTS: Inhabitants of the County of Funen (n = 471 732). MAIN OUTCOME MEASURES: Prevalence of potential drug interactions. RESULTS: One-third of the population were exposed to polypharmacy and among these 15% were exposed to drugs carrying a risk of harmful interaction. Among the elderly with polypharmacy, 25% aged 60-79 years and 36% over 80 years received drugs carrying the risk of interaction. Among individuals exposed to potential drug interaction, 62% were exposed only to one drug interaction and 38% to two or more different drug interactions. The drugs accounting for the highest number of potential interactions were diuretics, NSAIDs, ACE-inhibitors, digoxin, oral antidiabetics, calcium channel blockers, anticoagulants and beta-blockers. When focusing only on major drug interactions, potassium-sparing diuretics and oral anticoagulants were the most frequently involved drugs. CONCLUSION: Elderly patients exposed to polypharmacy should be kept under intensified monitoring as they are at increased risk of clinically significant drug interactions. PMID- 14531507 TI - Use of hypnotic and tranquillising drugs in general practice. Determinants, satisfaction and motivation to stop: the patients' view. AB - OBJECTIVE: To know more about the characteristics of long-term users of hypnotic and tranquillising drugs in a primary health care area in The Netherlands. DESIGN: A questionnaire was applied to users of benzodiazepines and the computerised medical records were analysed. SETTING: Nine general practices affiliated to the Registration Network Family Practices (RNH) of the University of Maastricht (UM). SUBJECTS: A sample of 506 users, aged > or = 30 years, was divided into four categories: short-term (< 3 months) users (once-only and episodic) and long-term users (episodic and continuous). RESULTS: In 1992/93, 81% were long-term users and 42% of these took their drug continuously. All groups investigated had a preponderance of women. Health problems more so than social problems led to use of the drug. Long-term use was associated with older age, depressive behaviour in handling problems, a higher score on the medicine addiction index, problems with children or housing, and satisfaction with the effects of the drug. CONCLUSION: Most people taking benzodiazepines are long-term users. Age, depressive behaviour, addiction, satisfaction and problems with children or housing are risk factors for long-term use. Gender is not risk factor. PMID- 14531508 TI - Medical doctors' perception of the "number needed to treat" (NNT). A survey of doctors' recommendations for two therapies with different NNT. AB - OBJECTIVE: While the number needed to treat (NNT) is in widespread use, empirical evidence that doctors or patients interpret the NNT adequately is sparse. The aim of our study was to explore the influence of the NNT on medical doctors' recommendation for or against a life-long preventive drug therapy. DESIGN: Cross sectional study with randomisation to different scenarios. SETTING: Postal questionnaire presenting a clinical scenario about a hypothetical drug as a strategy towards preventing premature death among healthy people with a known risk factor. Benefit after 5 years of treatment was presented in terms of NNT, which was set at 50 for half of the respondents and 200 for the other half. SUBJECTS: Representative sample (n = 1616) of Norwegian medical doctors. MAIN OUTCOME MEASURES: Proportion of doctors that would prescribe the drug. Reasons for recommending against the therapy. RESULTS: With NNT set at 50, 71.6% (99% CI 66.8-76.4) of the doctors would prescribe the drug, while the proportion was 52.3% (99% CI 47.5-57.1) with an NNT of 200 (chi = 50.7, p < 0.001). Multivariate logistic regression analysis indicated that the effect of NNT on the likelihood for recommending the therapy was age-dependent; young doctors ( < 36 of age) were more sensitive to the difference in NNTs than older doctors. Thirty-six percent (n = 464) of the doctors would not prescribe the drug, and 77.4% (99% CI 68.5 86.2) of those agreed with an argument stating that only one out of NNT patients would benefit from the treatment. CONCLUSION: Medical doctors appear to be sensitive to the magnitude of the NNT in their clinical recommendations. However, many doctors believe that only one out of NNT patients benefits from therapy. Clinical recommendations based on this assumption may be misleading. PMID- 14531509 TI - Structure and duration of consultations in Estonian family practice. AB - OBJECTIVE: To study the influence of age, gender and the nature of the patient's problems on length of time of consultation in the practices of newly trained family doctors in a recently reconstructed health care system. DESIGN: Video recordings of consultations with consecutive patients in family practice were studied for duration of consultation in relation to age, gender and nature of the problem(s). SETTING: Primary health care. SUBJECTS: 405 consecutive consultations were video-taped in the practices of 27 family doctors. MAIN OUTCOME MEASURES: Length of time of consultation and its segments was analysed using the Statistical Package for the Social Sciences. The problems were classified according to the ICPC. RESULTS: The average consultation lasted 9.0 min (+/- 4.9). Physical examination was 2.0 min (+/- 1.9) and was performed in 79% of all consultations. Respiratory and circulatory problems were the most common. More than one reason for the encounter was given in one-fourth of cases. Consultation time was longer for older age groups and for patients with psychological problems. CONCLUSION: Video-recording allows consultations to be evaluated directly and is acceptable to patients. The high participation rate of patients in our study can be explained by the individual approach and by the family doctor system. The period of consultation was dependent on patient age and on the number and nature of the problems, but was not influenced by gender. PMID- 14531510 TI - Characteristics of frequent attenders in different age and sex groups in primary health care. AB - OBJECTIVE: To compare frequent attenders in different age and sex groups with a control group according to characteristics and use of primary health care. DESIGN: Descriptive study of medical records. SETTING: Primary health care centre. SUBJECTS: 341 frequent attenders, divided by age and sex, were compared with 1025 controls from the same area. MAIN OUTCOME MEASURES: Consultation rate, diagnoses, prescribed medicine, referrals and certificates of illness during one year. RESULTS: Frequent attenders constituted 3.3% of the population but accounted for 25% of all visits to the GP and 44% of all illness certificates allocated. They were given more antibiotic prescriptions and consulted the GP more often for most medical problems. Frequent attenders had a morbidity, with a preponderance of infections at a young age, musculoskeletal and respiratory problems in middle age, and, among the oldest, circulatory, endocrine and musculoskeletal diseases. CONCLUSION: Frequent attenders had a morbidity within different ages similar to that of the controls, but also a higher consulting frequency for most medical problems and a significantly higher consumption of antibiotics. Are frequent attenders more vulnerable individuals and more prone to all kinds of diseases, or do they just consult more often for the usual medical problems? PMID- 14531511 TI - Change in the prevalence of coronary heart disease among Finnish elderly men and women in the 1990s. AB - OBJECTIVE: To characterise the prevalence of, and changes in, coronary heart disease (CHD) among men and women aged between 64 and 71 years in the 1990s. DESIGN: A study of clinical epidemiology involving two cohorts of elderly persons in 1990-1991 and 1998-1999. SETTING: Primary health care in the municipality of Lieto in southwestern Finland. SUBJECTS: Persons between 64 and 71 years of age in the southwest of Finland in 1990-1991 and 1998-1999. MAIN OUTCOME MEASURES: The occurrences of CHD were estimated using the history of a previous myocardial infarction or coronary revascularisation procedure evident in the medical records and with ischaemia or infarction as established on ECG according to the Whitehall criteria. RESULTS: The prevalence of 'probable' CHD decreased among men and women aged between 64 and 71 years, whereas the prevalence of 'possible' CHD decreased among women alone. Silent myocardial infarctions were common among women of both cohorts. Many more men of the second cohort, compared to the first one, had undergone a coronary angioplasty or bypass operation. CONCLUSION: The prevalence of CHD decreased among elderly women more clearly than among young elderly men. The favourable development illustrating a decrease in the prevalence of CHD among women should be sustained, while health promotion activities will need to be directed more actively towards men. PMID- 14531512 TI - Urinary incontinence in a Finnish population aged 70 and over. Prevalence of types, associated factors and self-reported treatments. AB - OBJECTIVE: To describe the prevalence, associated factors and self-reported treatments of different types of incontinence in a random older population. DESIGN: A population-based cross-sectional survey. SETTING: Third wave for the Tampere Longitudinal Study on Ageing (TamELSA) in 1999. SUBJECTS: 171 men and 227 women aged 70 and over. MAIN OUTCOME MEASURES: Urge and mixed incontinence combined and associated health and social indicators. RESULTS: The prevalence of stress, urge and mixed incontinence was 2%, 17% and 6%, respectively, in men and 23%, 6% and 30%, respectively, in women. In the logistic regression analyses adjusted for age, urge and mixed incontinence in women were associated with living in an institution (OR [odds ratiol 13.55), comorbidity (OR 1.94), ADL (activities of daily living) disability (OR 3.55), depressive mood (OR 2.49) and fecal incontinence (OR 7.84), while in men an association was found between urge and mixed incontinence and ADL disability (OR 2.59) and depressive mood (OR 2.69). All the men living in an institution were incontinent of urine. Of the incontinent women, 46% used diapers, but collecting devices were rare in men. CONCLUSION: The results emphasise the significance and multifactorial nature of the urge component in geriatric incontinence, which should be considered in both geriatric and general practice. PMID- 14531514 TI - Disaster waste management: a 'prestigious' subject. PMID- 14531513 TI - Child psychiatric networks of health centre doctors. AB - OBJECTIVE: To ascertain the personal networks GPs use when managing children with psychiatric problems. DESIGN: Cross-sectional postal study. SETTING: Health centres in the catchment area of Tampere University Hospital, Finland. SUBJECTS: GPs (n = 755) were sent a postal questionnaire. The response rate was 66.1%. MAIN OUTCOME MEASURES: A fill-in picture was used to identify professionals within the network of each GP. The quality of collaboration between GPs and professionals in health centres, other municipal authorities and secondary health care was assessed on a four-step Likert scale. RESULTS: A majority of respondents (64%) could name at least one person from their health centre with whom they cooperate in child psychiatric cases. The corresponding number in contact with other municipal authorities was 40%, and 25% with secondary care level. Almost all GPs (95%) had mostly positive experiences of cooperation with different professionals in the health centres, 73% with other municipal authorities and 47% with child psychiatric specialist care. CONCLUSION: The personal networks of health centre doctors call for improvement if the increasing child psychiatric problems are to be kept under control. This is a challenge for both primary and secondary care level doctors. PMID- 14531515 TI - Genetic algorithms as a promising tool for optimisation of the MSW collection routes. AB - Important advantages, including reductions in fuel consumption and labour cost, arise from the optimal design of solid waste (SW) collection routes. Further, optimal design can reduce vehicle maintenance expenditures and improve traffic conditions in urban areas. To date, optimal routes have been developed according to intuitive methodologies and field experience. However, increasing attention is being devoted to innovative approaches, such as those able to simulate complex collection systems. To analyse these complexities, operational research applications are used. They are typically based on the implementation of heuristic procedures allowing for high quality solutions to the problem at hand. From a computational point of view, however, heuristic procedures have a complexity which is o(n3), where n is the number of points which have to be visited during each route. This is a limit for an accurate representation of urban areas and for the quality of the calculated solutions. An alternative methodology, which is the subject of this paper, is based on a genetic algorithm. Also, an ad hoc algorithm, developed in the framework of a wider research, is illustrated. Results of a preliminary field test conducted for verification are also presented. PMID- 14531516 TI - Study on composition and particle characteristics of shredded municipal waste for the improvement of separation efficiency in a municipal bulky waste processing facility. AB - In order to propose a new and more efficient method of separation for shredded municipal bulky waste (SBW), particle characteristics (namely shape, density and terminal settling velocity) were investigated. It was found that particles of the combustibles such as wood, paper and plastics were included in the fraction smaller than 50 mm. Consequently, separation of SBW into combustibles and non combustibles by particle size is difficult. Since the particles of the various materials are of all shapes, it is difficult to sort them by shape discrimination. The overall separation efficiencies using air flow and density for SBW separation were estimated and the maximum values are as high as approximately 90% and 99% respectively. However, because of the overlapping ranges of the terminal settling velocities in air for glass, metals, rubber, and plastics, the overall efficiency will become smaller if those materials are major components of the feed. It is concluded that density separation is more suitable for the separation of combustibles and non-combustibles in SBW than air separation. PMID- 14531517 TI - Physical and chemical processes in baled waste fuel, with emphasis on gaseous emissions. AB - Over a period of seven months, the temperature and the emissions from six cylindrical and two rectangular stored bales containing waste for later use as fuel were measured. The bales were kept at two different ranges of temperatures: 30-35 degrees C and 20-25 degrees C respectively. It was found that only the rectangular bales showed significant production of CO2. The increase of emission for one of the rectangular bales kept at 30-35 degrees C, was 0.246% vol. CO2 per day for a 2-week period, after which there was a decrease of CO2 concentrations at a rate of 0.0224% vol. during a 32-week period. The other rectangular bale kept at 20-25 degrees C, exhibited a similar trend. However, the increase of CO2 concentrations was less at a rate of 0.0259% vol. per day during a 8-week period, after which the CO2 emission decreased at a rate of 0.0224% vol. per day during a 25-week period. All the bales exhibited aerobic decomposition in the sampling point. However on measuring the leachate concentrations, it was evident that the bales were actually in the equivalent acid-generating phase of a young landfill. The temperature inside the bales did not increase higher than the ambient air temperature. PMID- 14531518 TI - The recycling of MSW incinerator bottom ash by sintering. AB - In recognition of the trend toward an increased use of bottom ash as construction material, the authors have investigated the feasibility of recovering bottom ash for use as aggregates, by sintering size-fractioned MSW incinerator bottom ash (particle size less than 1.41 mm and between 4.76-1.41 mm) at 400-1,000 degrees C for 60-240 min, and then determining the sintered material characteristics, such as the compressive strength, heavy metal leachability and principal material properties. The results indicate that the pH of the Toxicity Characteristic Leaching Procedure (TCLP) leachate produced from both fine and the coarse ash, ranged from 10.0-11.5, and from 7.5-11.3 respectively, and showed a tendency to decrease with an increasing sintering temperature. In addition, for both types of ash the compressive strength of the sintered monoliths, ranging from 50-55 MPa, decreased slightly when the sintering temperature was increased from 400 to 600 degrees C. Deformation problems may arise from the melting of glassy substances in the ash when bottom ash is sintered at temperatures higher than 700 degrees C. Thus, when sintered between 800 to 1,000 degrees C, the sintered bottom ash pellets might disintegrate due to the formation of aluminium and/or calcium salts. The decomposition of calcium carbonate at 650 degrees C which releases significant amounts of carbon dioxide, may also cause the destruction of a monolith. Based on considerations of loss on ignition, volume changes, water adsorption, soundness, bulk density and the compressive strength of the sintered ash, developed by the sintering of bottom ash between 400 to 600 degrees C after removing its coarse impurities, the general results from the experiments suggest that the aggregates do meet the Chinese National Standards (CNS) for permeable blocks. PMID- 14531519 TI - A framework to contain a spatially distributed model of the degradation of solid waste in landfills. AB - This paper describes the concept and development of a generic spatially distributed numerical model that can contain and link sub-models of landfill processes in order to simulate solid waste degradation and gas generation in landfills. The model has been designed to simulate anaerobic degradation of solid waste saturated in leachate, and to reflect the effect on degradation of the control of both the flow and chemistry of the leachate in the pore space of the solids. The model can accommodate different profiles of waste constituents and therefore has the potential to simulate the effects of pre-treatment techniques such as shredding and inoculation. Components of the model simulate the transport of leachate and gases, and the consolidation of the solid waste. The results of some prototype calculations are presented. PMID- 14531520 TI - Determination of oxygen consumption for landfilled municipal solid wastes. AB - One of the serious problems of landfilling is the long time required for stabilisation. Nevertheless, site aeration proved to be effective to attenuate such problems. Oxygen consumption rate and air permeability/diffusion are the key parameters to set up the best allocation and design for vertical pipes to promote aeration and hence stabilisation of waste. This paper presents an investigation into the oxygen consumption for various landfilled wastes in Japan such as incineration ash, incombustible waste and under-sieve fraction of shredded bulky waste. Fresh waste sampled directly from processing facilities, as well as old waste sampled from landfill sites, were subjected to analytical determination and measurement of oxygen consumption. Bulky waste showed the highest oxygen consumption, however the old waste revealed a significant loss of biodegradability potential and lower oxygen consumption records. The oxygen consumption for short testing periods (10-days), tends to increase with the ignition loss (IL). The experimental data of respiratory tests (for old waste) were modelled and the fitting parameters are correlated to waste characteristics. PMID- 14531521 TI - Relating landfill gas emissions to atmospheric pressure using numerical modelling and state-space analysis. AB - Landfill gas (CO2 and CH4) concentrations and fluxes in soil adjacent to an old, unlined Danish municipal landfill measured over a 48-hour period during the passage of a low-pressure weather system were used to identify processes governing gas fluxes and concentrations. Two different approaches were applied: (I) State-space analysis was used to identify relations between gas flux and short-term (hourly) variations in atmospheric pressure. (II) A numerical gas transport model was fitted to the data and used to quantify short-term impacts of variations in atmospheric pressure, volumetric soil-water content, soil gas permeability, soil gas diffusion coefficients, and biological CH4 degradation rate upon landfill gas concentration and fluxes in the soil. Fluxes and concentrations were found to be most sensitive to variations in volumetric soil water content, atmospheric pressure variations and gas permeability whereas variations in CH4 oxidation rate and molecular coefficients had less influence. Fluxes appeared to be most sensitive to atmospheric pressure at intermediate distances from the landfill edge. Also overall CH4 fluxes out of the soil over longer periods (years) were largest during periods with rapidly decreasing atmospheric pressures resulting in emission of large amounts of CH4 during short periods of time. This effect, however, was less significant for the CO2 fluxes. PMID- 14531522 TI - Probabilistic assessment of contamination using the two-phase flow model. AB - A physically motivated model is indispensable for a successful analysis of the impact of leaching from nuclear waste storage sites on the environment and public health. While most analyses use the single-phase flow model for modelling unsaturated flow and solute transport, the two-phase flow model considering the resistance of gas to water flow is a more realistic one. The effect of the two phase flow model on the water content is theoretically investigated first in this study. Then, by combining a geostatistical generator using the turning bands method and a multi-phase transport code TOUGH2, an automatic process is used for Monte Carlo simulation of the solute transport. This stochastic approach is applied to a potentially polluted site by low-level nuclear waste in Taiwan. In the simulation, the saturated hydraulic conductivity is treated as the random variable. The stochastic approach provides a probabilistic assessment of contamination. The results show that even though water content from the two-phase flow model is only 1.5% less than the one from the single-phase flow model, the two-phase flow causes a slower movement but a wider lateral spreading of the plume in the unsaturated zone. The stochastic approach provides useful probability information which is not available from the deterministic approach. The probability assessment of groundwater contamination provides the basis for more informed waste management, better environmental assessment and improved evaluation of impact on public health. PMID- 14531523 TI - Hospital waste operational procedures: a case study in Brazil. AB - This study analysed the hospital waste management practices and their implications to public health in the city of Belo Horizonte, south eastern Brazil. Belo Horizonte is a regional capital city located 300 miles from the famous city of Rio de Janeiro, Brazil. It is only 100 years old and is the first city planned in Brazil. It was originally planned for 300,000 people but, today, it is home to more than 2 million people. This study assessed the operational procedures at five hospitals (170 to 450 beds) with different medical specialties in order to analyse their major waste problems. This study also gathered data on different types of treatment and disposal procedures used by waste generators, which enabled an estimate on the amount of on-site and off-site waste generated. PMID- 14531524 TI - A risk factor analysis of municipal domestic refuse landfills using a reactor with high water input. AB - Collapse, explosion and water pollution by leachate have been three major problems in confining the development of sanitary landfilling technology as an economical method for disposal of municipal refuse. Potential risk factors including settlement of landfilled refuse, the generation of methane, carbon dioxide and other gases from landfill, the dynamic changes in temperature within the landfill and its ambient environment, as well as COD(Cr), BOD5, nutrients (TKN, TP and organic matter), heavy metals (Cu, Pb and Zn) and pH in landfill leachate were simulated using a reactor with high water input. It was indicated that high content of organic components in landfilled refuse is a key risk factor to cause abrupt collapse and accidental explosion in the landfill because they have impact not only on the settlement, but also on the release of methane and other gases. Elevated temperatures coincided with maximum generation of methane and other gases, thus increasing the risk of accidental explosion. Leachate quality was closely related with the release of COD(Cr) and BOD5 and heavy metals such as Cu, Pb and Zn, as well as organic pollutants, there was less release of TKN and TP. The mass balance analysis for the reactor showed that 36% of organic carbon reduction in the refuse during the simulations was discharged into leachate as soluble organic carbon, and 3% was converted to methane and carbon dioxide as gas production. Mixing of refuse with soil and sand particles, the so called drill-a-hole method and recycling leachate were measures suggested in order to mitigate and avoid potential accidents. PMID- 14531525 TI - Clinical usefulness of virulence factors of Helicobacter pylori as predictors of the outcomes of infection. What is the evidence? PMID- 14531526 TI - Dysplasia in fundic gland polyps is associated with nuclear beta-catenin expression and relatively high cell turnover rates. AB - BACKGROUND: Fundic gland polyps (FGPs) occur in both syndromic and sporadic form. Syndromic FGPs arise through mutations in the adenomatous polyposis coli (APC) gene, whereas sporadic FGPs are caused by beta-catenin gene mutations. Dysplasia in sporadic FGPs, found less often than in syndromic FGPs, was recently associated with APC rather than beta-catenin mutations. These data suggest different functional consequences of APC and beta-catenin mutations. To investigate this hypothesis, we examined proliferative activity, degree of apoptosis, beta-catenin expression and p53 expression in syndromic and sporadic FGPs. METHODS: Syndromic FGPs (n = 9) from familial adenomatous polyposis (FAP) patients and sporadic FGPs (n = 18) were studied. Proliferative activity, apoptotic cell death and expression of beta-catenin and p53 were examined by immunohistochemistry. In FGPs containing dysplasia, areas with and without dysplasia were compared. RESULTS: Syndromic and sporadic FGPs without dysplasia exhibited similar proliferative activity, degree of apoptosis, beta-catenin and p53 expression. Dysplasia was observed more often in syndromic (4/9) than in sporadic FGPs (1/18). Within FGPs containing dysplasia, dysplastic areas showed abnormal nuclear beta-catenin staining in 3/5 cases and higher rates of cell proliferation and apoptosis than non-dysplastic areas. Overexpression of p53 was not observed. CONCLUSION: The finding of similar rates of proliferation and apoptosis in syndromic and sporadic FGPs does not support the hypothesis that APC and beta-catenin gene mutations have different functional consequences in FGPs. The association of dysplasia with relatively high cell turnover rates and nuclear expression of beta-catenin indicates activation of the Wnt-APC-beta-catenin pathway in dysplasia. The finding of dysplasia in some but not all syndromic FGPs suggests the involvement of other genes in addition to the APC gene in the development of dysplasia in FGPs. PMID- 14531527 TI - Helicobacter pylori, non-steroidal anti-inflammatory drugs and smoking in risk pattern of gastroduodenal ulcers. AB - BACKGROUND: Helicobacter pylori, NSAID and cigarette smoking are major risk factors for gastroduodenal ulcers. However, the results of studies on the interaction between these factors on ulcerogenesis are controversial. This study was designed to examine the association between gastroduodenal ulcers and H. pylori infection, NSAID use, smoking and age. METHODS: 5967 dyspeptic patients underwent 13C-urea breath test (UBT) and upper endoscopy, while age and dyspeptic symptoms were reported. RESULTS: Out of 5967 patients, 31.8% were ulcerated; 9.2% had gastric, 17.2% duodenal and 5.4% both gastric and duodenal ulcers. H. pylori was found in 72.5% of gastric ulcer patients, in 83.6% of duodenal ulcer patients, in 76.9% of gastroduodenal ulcer patients and in 64.8% of dyspeptic patients. The gastric, duodenal and gastroduodenal ulcers were related to H. pylori significantly and the respective ORs were: 1.44, 2.77 and 1.81. NSAID alone was used by 6.2%-12.7% of ulcer patients, tending to raise only the risk of gastric ulcer but reducing that of duodenal and gastroduodenal ulcers. The H. pylori prevalence was significantly higher in smokers (76%) than in non-smokers (67%) and the ulcer risk was also significantly higher in smokers than in non smokers. About 20% of ulcers were 'idiopathic', i.e. without NSAID and H. pylori and the ratio of these ulcers to all ulcers significantly increased during the 5 years of the study. CONCLUSIONS: Based on multivariable logistic regression analysis we conclude that: 1) H. pylori infection, NSAID use, smoking and age play major roles in the pathogenesis of peptic ulcerations; 2) there is a negative interaction between H. pylori and NSAID on duodenal ulcers, suggesting that H. pylori reduces the development of these ulcers in NSAID users, and 3) about 20% of peptic ulcers in the Polish population are unrelated to H. pylori and NSAID use (idiopathic ulcers). PMID- 14531528 TI - Duodenogastric reflux sustains Helicobacter pylori infection in the gastric stump. AB - BACKGROUND: Duodenogastric reflux (DGR) and Helicobacter pylori infection have been suspected of being contributing agents to the genesis of gastritis and subsequent cancer, but compelling, conclusive data about the exact relationship have been lacking. METHODS: We investigated the effect of DGR on H. pylori infection in 95 gastrectomized subjects divided into four groups according to type of reconstruction: the jejunal pouch interposition group (JPI, n = 36); the Roux-en-Y group (RY, n = 17); the Billroth I group (B-I, n = 20); and the Billroth II group (B-II, n = 22). The following items were examined for each group: the duration of DGR; the prevalence of H. pylori infection; other bacterial identification and quantity; and the severity of gastritis. RESULTS: The percent of total time of DGR was lower in the JPI (7%) and RY groups (28%) than in the B-I (59%) and B-II groups (88%) (P < 0.02). The prevalence of H. pylori infection was lower in the JPI (28%) and RY groups (29%) than in the B-I (60%) and B-II groups (73%) (P < 0.02). Inversely, the JPI and the RY groups had a higher quantity of other bacteria than the B-I group (P = 0.02). For all four groups, the stomachs infected with H. pylori were fewer than those tested negative for the organism (P < 0.0001). Inflammation scores were lower in both the JPI and RY groups than in the B-I and B-II groups (P < 0.05, respectively). CONCLUSIONS: Duodenogastric reflux facilitates the survival of H. pylori in the gastric stump after a distal gastrectomy. PMID- 14531529 TI - Changes in prevalence of Helicobacter pylori infection in two groups of patients undergoing endoscopy and living in the same region in the Netherlands. AB - BACKGROUND: There are few data on the prevalence of Helicobacter pylori in different populations and no data on changes in time. A cross-sectional study was done in a large population of patients undergoing endoscopy in order to relate the presence of H. pylori to ethnic background and to detect changes in time. METHODS: All consecutive patients over a 10-year period were included and biopsy specimens from the gastric antrum were taken for detection of H. pylori. A large population of immigrants living in the Zaanstreek region was studied separately. RESULTS: In the 10-year period, 14,909 consecutive diagnostic upper GI endoscopies were done in 11,691 consecutive patients. Only the first endoscopy done in a patient was included. Of 11,691 patients available for evaluation, there were 5669 men and 6022 women (mean age 55 years, range 4-99). Of these, 3201 (27.4%) were H. pylori-positive and 4988 (42.7%) were H. pylori-negative. No biopsy specimens were available for the remaining 3501 (29.9%) patients. In the group under study, 1309 (11%) patients were immigrants. H. pylori was present significantly more often in men and in immigrants, where prevalence was 75% versus 33% (P < 0.0001). The prevalence of H. pylori showed a marked decrease in both populations during the period of study. Extrapolating trend lines into the future indicates a prevalence of H. pylori below 10% by the year 2009. Among immigrants, this will be reached in approximately 2027. CONCLUSION: This study shows differences in prevalence of H. pylori in different populations. We found a decreasing prevalence that can be explained by the lower acquisition rate of the infection. It is important to be aware of the differences in prevalence of H. pylori in a population composed of different ethnicities. PMID- 14531530 TI - Necessity of multiple gastric biopsies from different sites for detection of clarithromycin-resistant Helicobacter pylori strains. AB - BACKGROUND: It has been reported that approximately 10% of patients infected with Helicobacter pylori have both clarithromycin-susceptible and clathromycin resistant strains. However, there have been no reports indicating whether only one gastric biopsy is sufficient to detect clarithromycin-resistant strains. METHODS: Sixty-five H. pylori-infected patients were selected for this study, and 40 of them were given clarithromycin-based eradication therapy. Four gastric biopsies, 2 from the antrum and 2 from the corpus, were obtained from each of the 65 patients. Susceptibility of H. pylori strains to clarithromycin was examined by detecting mutations of the 23S ribosomal RNA (rRNA) gene of H. pylori. RESULTS: The clarithromycin-resistant strains were detected in 16 of the 65 (25%) patients. Only 5 of the 16 (31%) patients had the resistant strains in both the antrum and corpus. When only 1 or the other biopsy from the antrum was used, the resistant strains were detected in 8 (50%) or 9 (56%) of the 16 patients. CONCLUSIONS: These data indicate that multiple gastric biopsies from both the antrum and the corpus should be used to detect clarithromycin-resistant H. pylori strains. PMID- 14531531 TI - An irritable bowel syndrome-specific symptom questionnaire: development and validation. AB - BACKGROUND: No self-assessment instruments are available to assess symptoms of irritable bowel syndrome (IBS). Our aim was to develop a disease-specific symptom questionnaire for use in patients with IBS. METHODS: Two-hundred-and-thirty-four patients (77% F) with a mean age of 44 years took part in a psychometric evaluation using the previously validated Gastrointestinal Symptom Rating Scale modified for use in patients with IBS (GSRS-IBS). This version was tested against several disease-specific health-related quality of life (HRQL) questionnaires. Items with a high ceiling effect, items that measured a different construct, and items showing high correlation (>0.80) to another item were removed. A confirmatory factor analysis was also performed. RESULTS: The final questionnaire included 13 items depicting problems with satiety, abdominal pain, diarrhoea, constipation and bloating. The internal consistency reliability was high, ranging from 0.74 (pain) to 0.85 (satiety). The associations between similar constructs in the GSRS-IBS and the various HRQL scores confirmed the construct validity. Pain, bloating and diarrhoea were the symptom clusters that impaired HRQL the most. CONCLUSION: The GSRS-IBS is a short and user-friendly instrument with excellent psychometric properties. PMID- 14531532 TI - Nitric oxide has tonic inhibitory effect, but is not involved in the vagal control or VIP effects on motility of the porcine antrum. AB - BACKGROUND: The involvement of nitric oxide (NO) in vagal control and vasoactive intestinal polypeptide (VIP)-induced effects on antral motility was studied using isolated perfused preparations of porcine gastric antrum with intact vagal innervation. METHODS: The presence of NO and VIP-producing neurons was studied using immunohistochemistry and histochemical techniques. Widespread, but not total, co-localization of NO and VIP immunoreactivity was found in the submucosa and in the muscle layers. RESULTS: Electrical stimulation of the vagus nerves for 5 min (8 Hz, 10 mA, 4 msec) increased the motility index from 2.47 = 0.44 to 11.50 +/- 2.02 (n = 5). This effect was not influenced by the two NO synthase inhibitors N-nitro-L-arginine methyl ester (10(-4) M) and NG-nitro-L-arginine (10(-5) M). However, infusion of inhibitors increased the spontaneous motility index from 2.40 +/- 0.08 to 5.36 +/- 1.08 (P < 0.05) and 3.05 +/- 1.10 to 4.14 +/ 1.04 (P < 0.05), respectively. The addition of L-arginine reversed this effect. Infusion of VIP 2 x 10(-9)M decreased the motility index from 2.32 +/- 0.43 to 1.32 +/- 0.27 (P < 0.05), an effect that was preserved during NO synthase inhibition. Electrical vagus stimulation increased the release of VIP to the venous effluent, an effect that persisted during NO synthase inhibitors. CONCLUSION: We conclude that NO-producing nerves seem to have a tonic inhibitory action on the porcine antral motility, but are not involved in the motor effects of vagal stimulation or VIP infusion. PMID- 14531533 TI - Percutaneous endoscopic gastrostomy (PEG) for enteral nutrition in patients with stroke. AB - BACKGROUND: The use of percutaneous endoscopic gastrostomy (PEG) for enteral nutrition in patients admitted for stroke is difficult, varying and needs specific consideration. There is therefore need for more data on this patient group. We examined the indications, survival, tube removal and time with PEG in stroke patients and in other patients with PEG with the aim of providing guidance for the management of enteral nutrition via PEG in stroke patients. METHODS: Retrospective assessment of data from all stroke patients and patients with other diseases (control group) who had received PEG for enteral nutrition during a period of 8.5 years. RESULTS: Eighty-three stroke patients with dysphagia received PEG after unsuccessful use of nasogastric tubes or long-term tube feeding. Early mortality rate was 19% in the stroke group, 26% in the older group (>74 years) and 12% in the younger group (60-74 years). The PEG tubes were later removed due to swallowing recovery in 20% of the older group and in 31% of the younger group. At 90 days, 50%-60% still needed PEG. The stroke patients were older compared to the control group (n = 115); 30-day mortality was similar but more patients recovered the ability to swallow. CONCLUSIONS: Stroke patients are older than other patients who receive PEG; 27% have swallowing recovery and more than 75% have long-term need for PEG. Nasogastric tubes often fail, and the need for early PEG placement (within 2 weeks) must be assessed in appropriate patients. The patient's prognosis, the objective of nutritional treatment, duration of dysphagia, age and comorbidity should all be taken into consideration. PMID- 14531534 TI - In vivo sugar diffusion in the ileal epithelium of spontaneously hypertensive rats. AB - BACKGROUND: D-glucose absorption, distinguishing between active and diffusive components, was studied in the ileum of spontaneously hypertensive rats (SHR) and their normotensive control Wistar-Kyoto (WKY) rats. Net water transport was also determined. METHODS: A perfusion system in vivo with ileum loops was used, and experiments with phlorizin, phloretin and 2,4,6-triaminopyrimidine (TAP) were performed in order to discriminate between active and diffusive components and between transcellular and paracellular routes. RESULTS: A significant decrease in total D-glucose absorption was found in SHR compared to WKY rats, this reduction being due to a lower SGLT1-mediated component. The effect was not compensated by the total diffusive component, since the phlorizin-insensitive D-glucose absorption did not significantly change between rat strains. However, the diffusive component of D-glucose transport was relatively more important in hypertensive than in normotensive rats. The use of 2,4,6-triaminopyrimidine (TAP), which blocks the transport across the paracellular route, showed that the paracellular diffusion of D-glucose was higher in SHR than in WKY rats. Intestinal net water absorption was not modified between either group of animals, though the presence of phlorizin in the perfusate decreased the ileal water absorption to a greater extent in normotensive rats. CONCLUSION: The observed reduction in D-glucose absorption in vivo in the ileum of SHR was due to a decrease in the SGLT1-active component. Despite the paracellular diffusion of D glucose being higher in hypertensive than in normotensive rats, the total diffusion component was not high enough to compensate this alteration. PMID- 14531535 TI - Pharmacological monitoring of azathioprine therapy. AB - BACKGROUND: Studies on azathioprine (Aza) treatment in Crohn disease have indicated a positive correlation between clinical remission and a concentration in erythrocytes of the metabolites 6-thioguanine nucleotides (E-6-TGN) above 230 pmol/8 x 10(8) RBC. A concentration of the methylated Aza metabolites (E-6-MMP) above 5000 pmol/8 x 10(8) RBC has been correlated to hepatotoxicity. Thiopurine methyltransferase (TPMT) is responsible for the formation of methylated metabolites and lower E-TGN levels, and TPMT genotyping has been proposed as guidance for dosage. In a cross-sectional study we investigated relationships between the clinical outcome and Aza dose, the TPMT genotype and the Aza metabolite levels among patients with Crohn disease. METHODS: TPMT genotype (PCR assay), azathioprine metabolite levels (HPLC analysis) and xanthine oxidase (XO) activity were determined once in 71 randomly selected Crohn patients on an unaltered Aza dose for at least 3 months. RESULTS: None of the doses of Aza, TPMT genotype, E-6-TGN-, E-6-MMP levels or XO activity were significantly related to disease activity (H-B score), (P = 0.18, P = 0.69, P = 0.90, P = 0.54, P = 0.29, respectively). Leucopenia and/or hepatotoxicity were not demonstrated in any patient. Four patients had a heterozygous TPMT genotype (6.1%; 95% CI: 1.68% 14.80%). The 4 TPMT heterozygous patients had higher E-6-TGN levels than did the 67 remaining patients (P = 0.008). CONCLUSIONS: To explore the applicability of TPMT genotyping, E-6-TGN and E-6-MMP levels for therapeutic drug monitoring, large prospective studies with patient entry at the start of Aza therapy are needed. Until the results of such studies are available, the dose adjustments of Aza should be guided primarily by clinical response and blood counts; metabolite level measurements can only be applied to identify therapeutic non-compliance. PMID- 14531536 TI - What concerns subjects with inflammatory bowel disease and an ileostomy? AB - BACKGROUND: The greatest concern of patients with inflammatory bowel disease (IBD) is of having an ileostomy. The aim of this study was to describe worries and concerns in subjects with IBD and an ileostomy, and aspects of quality of life and coping strategies. METHODS: 21 subjects with an ileostomy were included, mean age 51, range 36-65 (F/M = 12/9), Crohn disease (CD) n = 14, ulcerative colitis (UC) n = 6 and indeterminate colitis (IDC) n = 1. Worries and concerns were assessed using the rating form of IBD patient concerns (RFIPC). Health related quality of life (HRQOL) was assessed using Short Form 36 (SF-36) and compared with a matched group for age and gender from the general population. Subjects' definition of quality of life, as well as perceived quality of life on a visual analogue scale, was evaluated. Coping strategies were investigated using the Jalowiec coping scale (JCS 40). RESULTS: Greatest concerns were related to intimacy, access to quality medical care, energy level, loss of sexual drive, producing unpleasant odours, being a burden, ability to perform sexually, attractiveness and feelings about the body. Vitality was significantly reduced compared to controls. Subjects' definition of good quality of life mainly concerned social dimensions of life and health. Low values on perceived quality of life indicated greater concerns. Confrontational coping style was most frequently used. CONCLUSION: The greatest concern for subjects with an ileostomy was intimacy. Vitality was reduced compared to controls. Integrating items of concern into counselling may result in greater coping ability and improved quality of life. PMID- 14531537 TI - Acute cholecystitis: delayed surgery or observation. A randomized clinical trial. AB - BACKGROUND: The aim of the present study was to compare the risk of observation versus that of cholecystectomy in acute cholecystitis in patients randomly allocated to delayed operation or conservative treatment. METHODS: One-hundred and-eighty patients were considered for participation in the study; 71 were excluded according to predefined criteria and 45 did not join for other reasons. The remaining 64 patients were randomized to cholecystectomy (n = 31) or observation (n = 33). Randomized patients were contacted regularly and followed up for a median of 67 months. All gallstone-related hospital contacts were registered in both randomized and excluded patients. RESULTS: Gallstone-related complications or emergency admissions for pain occurred in six patients in the operation group (19%; 95% CI 5%-33%) and in 12 patients (36%; 9% CI 20%-53%) in the observation group. Twenty-seven of 31 patients randomized to cholecystectomy had a cholecystectomy at a median of 3.6 months after randomization, and, of these, 3 (11%; 95% CI 0%-23%) suffered a major and 7 (26%; 95% CI 9%-42%) a minor complication. Ten patients randomized to observation later had their gallbladders removed, 1 (10%; 95% CI 0%-29%) patient had a major and 1 (10%; 95% CI 0%-29%) a minor complication. We found no mortality after cholecystectomy. CONCLUSIONS: We found a certain risk of subsequent gallstone-related events following conservative treatment of acute cholecystitis, but the data also show that cholecystectomy should not necessarily be compulsory after acute cholecystitis. PMID- 14531538 TI - Long-term effect of corticosteroid treatment in primary sclerosing cholangitis patients. AB - BACKGROUND: A beneficial effect of corticosteroids in primary sclerosing cholangitis (PSC) has been suggested, but characteristics of responding patients and long-term outcome have not been assessed. In this retrospective study, we aimed to characterize PSC patients selected for corticosteroid treatment at our centre and to identify potential factors associated with response. METHODS: We first compared groups of PSC patients treated (n = 47) and not treated (n = 88) with corticosteroids. Responding (n = 20) and non-responding (n = 27) patients were subsequently compared. Complete and partial responses were defined according to criteria established for autoimmune hepatitis. A third response category included improvement of symptoms and at least 50% reduction of transaminase and/or bilirubin levels during the first 6 months. RESULTS: At diagnosis of PSC, patients treated with corticosteroids were significantly younger, had higher serum levels of alanine transaminases, and more histological features of autoimmune hepatitis compared to the non-treated group. Complete treatment response was obtained in three patients and partial response in two, together comprising 3.7% of all PSC patients in this study. Fifteen patients fulfilled criteria of the third response category. Response to treatment was associated with higher serum levels of alanine transaminases and bilirubin and lower levels of alkaline phosphatases at treatment start. Responders had better long-term survival than non-responders (hazard ratio 6.28; 95% confidence interval 1.62 to 24.4; P = 0.008). CONCLUSIONS: A subgroup of PSC patients seems to respond favourably to corticosteroid treatment and may obtain improved long-term survival. PMID- 14531540 TI - Non-endoscopic first-line treatment of bleeding peptic ulcer with ranitidine and tranexamic acid. PMID- 14531539 TI - Clinical significance of anti-multiple nuclear dots/Sp100 autoantibodies. AB - BACKGROUND: Autoantibodies against discrete variable-sized dots observed in HEp2 cells by indirect immunofluorescence (IIF) test, called multiple nuclear dots (MND), have been closely associated with primary biliary cirrhosis (PBC). Some authors have argued that this antibody is also present in connective tissue diseases or liver diseases other than PBC as autoimmune chronic active hepatitis, particularly of the cholestatic type. We studied an unselected group of patients routinely tested for autoantibodies and positive for the MND pattern and tried to establish the correlation between the presence of this antibody and their diagnosis. METHODS: A commercial ELISA test, using a recombinant 26 kD truncated sequence of the Sp100 protein, corresponding to an immunodominant molecular region, was used to assess the clinical correlation of these autoantibodies in 110 patients showing an anti-MND immunofluorescence pattern. RESULTS: One-hundred and-ten patients were MND positive by IIF. Of these, 100 were Sp100 positive by ELISA. In the Sp100 positive group, 34 had a diagnosis of PBC (30 definite and 4 suspected) while 15 patients had a non-PBC hepatopathy. Unexpectedly, 13 of the MND/Sp100 positive pattern corresponded to systemic lupus erythematosus (SLE) patients and 5 cases to collagen diseases. Another divergence with previous reports was that 34 of the positive patients showed very heterogeneous clinical pictures, different from hepatopathies or collagen diseases. CONCLUSIONS: Anti Sp100 antibodies can be found in many clinical conditions. Testing for MND/Sp100 positivity is useful for the diagnosis of PBC, but only when the right clinical context is present. Other diseases cannot be excluded in first line SLE. PMID- 14531541 TI - Redox imbalance in the colonic mucosa of ulcerative colitis. PMID- 14531542 TI - Carcinoid tumour of the terminal ileum simulating Crohn disease. AB - Carcinoid tumours are often difficult to diagnose because of obscure or non specific symptoms. Two cases of ileal carcinoid are reported in whom the diagnosis was delayed as the symptoms and small-bowel series were thought to be consistent with Crohn disease. This report emphasizes the difficulties of diagnosing carcinoid by conventional radiological methods. Ileal carcinoid should be considered in the differential diagnosis of Crohn disease, particularly in elderly patients presenting with chronic recurrent symptoms. PMID- 14531543 TI - Human intraperitoneal microdialysis: increased lactate/pyruvate ratio suggests early visceral ischaemia. A pilot study. AB - BACKGROUND: Previous studies suggest that visceral ischaemia precedes shock and multiple organ failure, though methods for studying humans are lacking. We aimed to evaluate intraperitoneal microdialysis, a new technique for detecting splanchnic ischaemia in clinical practice. METHODS: Right-sided hemicolectomy was performed in eight patients who were studied by microdialysis postoperatively for glucose, lactate, pyruvate and glycerol levels. RESULTS: Six of the eight patients showed a normal postoperative course and had lactate/pyruvate ratios between 7.1 and 21.7, glucose between 4.5 and 14.3 mmol/L and glycerol between 10.4 and 296 micromol/L. In one patient, intraperitoneal lactate/pyruvate ratio and glycerol increased and glucose decreased 5 h before low oxygenation appeared. Another patient exhibited a period of increased lactate/pyruvate ratio before a period of atrial fibrillation. CONCLUSION: Intraperitoneal microdialysis was performed safely. Two out of the eight patients exhibited changes of metabolic markers followed by clinical symptoms that were probably related to transient visceral ischaemia. Our findings suggest that intraperitoneal microdialysis may become a useful tool for monitoring splanchnic ischaemia in clinical practice. PMID- 14531544 TI - Differences in the density and spatial distribution of florid and diffuse plaques in variant Creutzfeldt-Jakob disease (vCJD). AB - OBJECTIVE: To determine whether in cases of variant Creutzfeldt-Jakob disease (vCJD), the florid-type plaques are derived from the diffuse plaques or whether the 2 plaque types develop independently. MATERIAL: Blocks of frontal, parietal, occipital and temporal neocortex and cerebellar cortex from 11 cases of vCJD. METHOD: The density, distribution and spatial pattern of the florid and diffuse plaques were determined in each brain region using spatial pattern analysis. RESULTS: The density of the diffuse plaques was significantly greater than that of the florid plaques in most areas. The ratio of the diffuse to florid plaques varied between brain regions and was maximal in the molecular layer of the cerebellum. The densities of the florid and diffuse plaques were positively correlated in the parietal cortex, occipital cortex, the inferior temporal gyrus and the dentate gyrus. Plaque densities were not related to disease duration. In the cerebral cortex, the diffuse plaques were more commonly evenly distributed or occurred in large clusters along the cortex parallel to the pia mater compared with the florid plaques which occurred more frequently in regularly distributed clusters. CONCLUSION: The florid plaques may not be derived from the diffuse plaques, the 2 plaque types appearing to develop independently with unique factors involved in their pathogenesis. PMID- 14531545 TI - Alpha-internexin expression in medulloblastomas and atypical teratoid-rhabdoid tumors. AB - OBJECTIVE: To determine whether medulloblastomas and atypical teratoid/rhabdoid tumors express alpha-internexin, an intermediate filament protein that is expressed in normal neurons undergoing maturation and differentiation. MATERIALS AND METHODS: 28 medulloblastomas and 5 atypical teratoid/rhabdoid tumors were examined for the immunohistochemical expression of alpha-internexin, as well as the neuronal markers peripherin and synaptophysin, and glial fibrillary acidic protein. RESULTS: Overall, 21 of 28 medulloblastomas (75%) expressed alpha internexin. More specifically, alpha-internexin expression was observed in 6 of 10 (60%) classic medulloblastomas, 12 of 14 (86%) desmoplastic medulloblastomas, 2 of 3 (67%) nodular medulloblastomas, and in one medullomyoblastoma. Similarly, 4 of 5 (80%) atypical teratoid/rhabdoid tumors expressed alpha-internexin. The extent of staining for alpha-internexin tended to be less than that of synaptophysin for both medulloblastomas (75% vs 93%) and atypical teratoid/rhabdoid tumors (80% vs 100%). In contrast to alpha-internexin, peripherin was expressed in only 4 medulloblastomas and one atypical teratoid/rhabdoid tumor. CONCLUSIONS: Alpha-internexin is expressed in the majority of medulloblastomas and atypical teratoid/rhabdoid tumors, indicating that these primitive tumors usually exhibit neuronal differentiation. PMID- 14531546 TI - Novel crystalloid structures in suprasellar paraganglioma. AB - A-52-year-old woman was admitted to a hospital because of 2-year history of abnormal behavior and impaired visual acuity. Magnetic resonance imaging delineated a sizable mass at the suprasellar region. The partially removed tumor was arranged in irregular lobules composed of an admixture of clusters of cobblestone-like small cells and process-bearing cells with ovoid nuclei, surrounded by a fine, neuropil-like matrix. The Zellballen structure was inconspicuous, and mitosis was absent. Immunohistochemically, the tumor cells were positive for chromogranin A, synaptophysin, class III beta-tubulin and neurofilament, while negative for glial fibrillary acidic protein, cytokeratin and all 6 pituitary hormones. S100 protein expression was limited to cells adjacent to stroma. The MIB-1 labeling index was 0.5%. Histopathological diagnosis was paraganglioma of abortive architecture. Ultrastructurally, numerous dense-cored vesicles were found within the processes and cytoplasm. Synapse formation was not demonstrated. Interestingly, crystalloids up to 3 microm in size were frequently found. They had hexagonal or quadrilateral architecture without limiting membranes. The interval between periodically arranged fibrils was variable, ranging from approximately 20 - 50 nm. Retrospective examination by light microscopy failed to reveal corresponding structures. Crystalloids are rare manifestation of paragangliomas, yet undescribed in those of intracranial origin. Furthermore, the ultrastructure of the present case differs from those of previous cases. PMID- 14531547 TI - MIB-1 immunoreactivity in craniopharyngiomas: a clinico-pathological analysis. AB - BACKGROUND: Craniopharyngiomas are epithelial tumors of the suprasellar region, unanimously defined as benign. Despite the benign histological aspect and apparent gross total resection achieved in a proportion of cases, recurrence rate remains about 30% at 10 years. The role of 7 pathological factors as predictors of recurrence and clinical outcome in craniopharyngioma patients is controversial, as well as the prognostic role of the evaluation of proliferation potential. MATERIAL AND METHODS: In the present study, the proliferation potential of 37 craniopharyngiomas was investigated by analyzing the pattern of Ki-67-MIBI immunoreactivity; the data have been analyzed in relation to age of the patient, histologic type (adamantinomatous/squamous papillary), microscopic and cytochemical features. RESULTS: Craniopharyngiomas operated in adults have a higher MIB-1-LI than those of children; the epithelial cells abutting the stromal cysts are engaged in the cell cycle, while this is not the case for the cells abutting the nervous tissue; MIB-1-LIs of adamantinomatous craniopharyngiomas are not different from those of squamous papillary craniopharyngiomas, the localization of MIB-1-positive nuclei is different in the 2 craniopharyngioma types, non-epithelial cells proliferate in the stromal component of craniopharyngiomas. CONCLUSIONS: By analyzing these data while considering all available information on the efficacy of surgery and radiotherapy, the proliferation potential of craniopharyngiomas, when evaluated by MIB-1, has no role in the outcome. The very low MIB-1-LI of the intracerebral tumor growth indicates that uncompletely resected tumor remnants do not represent an active starting point of recurrence. PMID- 14531548 TI - "Tumor-mimicking" multiple sclerosis. AB - The spectrum of clinical manifestations of multiple sclerosis (MS) may include rare cases where cerebral lesions simulate brain tumors or abscesses on neuroimaging. We report here on a 43-year-old woman with numerous ring-enhancing cystic lesions in the white matter of cerebral hemispheres, brainstem and cerebellum. The radiological picture was overwhelmingly in favor of a metastatic or infectious etiology, but brain biopsy showed subacute demyelination with central necrosis. PMID- 14531549 TI - Is there a final common pathway in mitochondrial encephalomyopathies? Considerations based on an autopsy case of Kearns-Sayre syndrome. AB - A case of Kearns-Sayre syndrome (KSS) diagnosed 18 years prior to death due to stroke and heart failure with postnatal onset was followed over 15 years and confirmed by postmortem examination. Within the brain, an old cystic infarction of the left hemisphere was found. Other features included white matter gliosis and cerebellar atrophy. Equal or similar features were observed in other conditions thought to be due to failure of mitochondrial metabolism, therefore, a common evolution of neuropathological changes must be discussed. PMID- 14531550 TI - Extracerebral subdural manifestation of Chester-Erdheim disease associated with a giant adenoma of the pituitary. AB - Chester-Erdheim disease is a rare non-Langerhans histiocytosis, affecting within the CNS mainly the neurohypophyseal unit, the retrobulbar space and the parenchyma of cerebellum, cerebrum and brainstem. Here we present a case of a 55 year-old woman who developed an exophthalmus, edema and dyspnea, finally leading to death 4 months post admission to the hospital. A cMRI showed a retrobulbar fibrosis, a tumor in the sella turcica, and further tumor formation expanding from the pons to the spinal cord, but without involvement of the dural sheet. Autopsy revealed multiple tumors attached to the pituitary gland, the tentorium, and the brainstem as well as a diffuse thickening of the dura. Histologically, the tumor tissue consisted of densely packed lipid-laden foamy macrophages positive for CD68 and intervening fibrillary cords. Interestingly, tumor cells did not infiltrate/affect the parenchyma but showed a strictly extracerebral/ subdural location. In addition, sections of the pituitary tumor revealed a chromophobe giant adenoma of the pituitary gland. As to our knowledge this is the first detailed description of an exceptional case of intracranial CED presenting with strictly extracerebral/subdural tumor masses accompanied by a giant adenoma of the pituitary gland. PMID- 14531551 TI - Reinduced expression of developmental proteins (nestin, small heat shock protein) in and around cerebral arteriovenous malformations. AB - Intermediate filament (IF) nestin and small heat shock protein (sHSP) are developmentally regulated proteins. Nestin is highly expressed on proliferating neuroepithelial stem cells of the developing central nervous system (CNS). During the developmental neurulation stage, nestin is replaced by mature neuronal (neurofilament) or glial cell-specific IFs (glial fibrillary acidic protein, GFAP). Several pathologic states induce astrocytes to synthesize nestin transiently in the mature brain. However, the exact nature of the embryonic conversion from nestin to mature cytoskelton is unclear. In an attempt to define the effect of ischemic hemodynamic stress caused by cerebral arteriovenous malformation (AVM) on the brain parenchyma, we examined the synthesis and cellular distribution of sHSP and nestin in vascular elements of AVMs and in the gliotic area surrounding AVMs. Ten consecutively collected surgical specimens meeting the histological criteria for AVM were immunohistochemically stained using primary antibodies for nestin, HSP27 and alphaB-crystallin. Nestin, HSP27 and alphaB-crystallin mRNA expressions were examined by reverse transcriptase polymerase chain reaction (RT-PCR). Nestin expression is reinduced not only in reactive astrocytes, but also in endothelial cells in the surrounding gliotic tissue of the cerebral AVM. These cells also expressed sHSP (HSP27, alphaB crystallin) that maintain the integrity of the IF network and prevent unfolding of cellular proteins induced by various stresses. RT-PCR showed the increased expression of sHSP and nestin mRNA in the AVM specimens. These results indicate that embryonic reversion of the mature cytoskeleton to nestin and the increased expression of sHSP in response to cerebral injury are associated with increased wall tension caused by dilating AVM vessels and with the hemodynamic stress that surrounds AVMs. PMID- 14531552 TI - Editors' remarks: Evolving concepts for local therapies for glioma. PMID- 14531553 TI - Intraoperative imaging--MRI. AB - Neuronavigation has become a standard technique in many neurosurgical procedures where its use allow better positioning of the craniotomy flap, precise targeting of lesions, and better anatomical orientation. However, the imaging used in such procedures is acquired preoperatively and thus, cannot project the dynamic changes that occur during surgery and result in many cases in significant brain shift and decreased accuracy. Recent technological developments have yielded a variety of MRI machines that can be used intraoperatively and provide the surgeon with updated images, integrated navigation capabilities, full compensation for brain shifts, and the ability to assess the extent of resection of the lesion. The concepts behind such technologies vary from one manufacture to another resulting in systems that vary in complexity, ease of use, spatial demands, and cost. In this chapter we review our experience with two intraoperative MRI systems used in a variety of neurosurgical procedures: the GE Signa SP System and the Odin PoleStar System. PMID- 14531554 TI - Stereotactic volumetric resection of gliomas. AB - The aim of stereotactic volumetric resection of intrinsic brain tumors at the time of design of the method was a most radical and complete resection of all tumor tissue while reducing morbidity by using minimally invasive approaches. This should also using the precision and accuracy of stereotaxis allow for resection of deep-seated tumors previously believed to be unresectable. Numerous retrospective studies have been performed and have shown that radical resection is feasible using this methodology and even so in eloquent brain areas. Whereas in malignant gliomas there is no proof of increased survival or time-to progression after stereotactic volumetric resection quite favourable results have been obtained in deep-seated low grade gliomas. What the actual role of this modality is in comparison to other forms of local treatment of circumscribed CNS lesions remains to be open in view of the lack of comparative studies. PMID- 14531555 TI - Fluorescence-guided resections of malignant gliomas--an overview. AB - Radical resections of contrast-enhancing tumour in patients with malignant gliomas may be pertinent for survival but are often difficult to achieve due to uncertainties in distinguishing tumour margins intra-operatively. In this respect a number of novel methods are being examined which aim at enhancing resections. Among these methods, resections that exploit the accumulation of fluorescent porphyrins within malignant glioma tissue in response to exogenous administration of a metabolic percursor, 5-aminolevulinic acid, may offer particular advantages. This article summarises the clinical background and current status of 5-ALA drug development for fluorescence-guided resections of malignant gliomas and analyses the available literature with regard to possible mechanisms that govern the highly specific accumulation of fluorescent porphyrins in malignant glioma tissue in response to 5-ALA administration. PMID- 14531556 TI - Postoperative imaging after brain tumor resection. AB - Prognosis after surgery for a brain tumor is not only influenced by its histology but also by the completeness of resection. The exact identification of a possible residual tumor is crucial also for follow-up and the purpose of evaluation of response to treatment. A review of the current and past literature addressing the individual value, the technique, and the pitfalls of postoperative imaging with Magnetic Resonance Imaging (MRI) and Computed Tomography (CT) for the definition of a postoperative residue after the resection of brain tumors is given. The different problems and possibilities are illustrated by imaging examples. PMID- 14531557 TI - Preliminary experience in glioma surgery with intraoperative high-field MRI. AB - OBJECTIVE: To apply a new setup, combining the benefits of high-field magnetic resonance imaging (MRI) with microscope-based neuronavigation, providing anatomical and functional guidance, in glioma surgery. MATERIAL AND METHODS: MR imaging was performed using a 1.5 T scanner, located in a radiofrequency-shielded operating theatre. The patient is lying on a rotating operating table, which is locked at the 160 degree position for surgery at the 5 G zone and turned into the scanner for imaging. The microscope, placed in the 5 G zone, in combination with a ceiling mounted navigation system enables microscope-based neuronavigation; integrated data from magnetoencephalography and functional MRI provide functional guidance. RESULTS: 126 patients were investigated with intraoperative high-field MRI, among them were 37 patients with gliomas. In the biopsy/catheter group (n = 8) MRI reliably depicted the needle position or the location of catheter placement. In the group with glioma resection (n = 29) intraoperative MRI revealed that the surgical objective was not achieved in 28%, leading to further tumour removal. We did not observe complications attributable to intraoperative high-field MRI. Image quality was not diminished by the operating room equipment, so that there was nearly no noticeable difference between pre- and intraoperative image quality. Neuronavigational guidance was applied in 31 patients: the integrated use of functional data prevented an increased morbidity despite extended resections. CONCLUSION: Intraoperative high-field MRI allows a reliable delineation of the extent of resection in glioma surgery. If the surgical objective was not met, a modification of the surgical strategy during the same operation is possible, thus leading to more radical resections. Furthermore, high field MRI offers increased image quality and a much broader spectrum of different imaging modalities, compared to previous intraoperative low-field systems. PMID- 14531558 TI - Awake craniotomy in glioma surgery. AB - Awake craniotomy is the cheapest and most reliable method to ensure neurological integrity in cerebral gliomas that infiltrate or come close to the eloquent areas of the brain, allowing (a) the localization of eloquent cortical areas by electrical stimulation and epileptic foci by cortical recording, and (b) the monitoring of the functional integrity of awake patients while aiming at subtotal removal of the gliomatous tissue. In addition, awake craniotomy opens a brief but unique window to the living brain for (a) basic neuroscience, including verification of preoperative functional imaging data and recording of electrophysiological correlates of mental processes, and for (b) applied research, including development of innovative instrumentation for brain recording and monitoring as well as screening for potential areas to be modulated in movement disorders and chronic pain. PMID- 14531559 TI - External beam and conformal radiotherapy in the management of gliomas. AB - External beam radiotherapy remains an important local treatment modality in both high and low grade gliomas, however its contribution to outcome remains modest. In high grade tumours this is because of their extreme clinical radioresistance, with local recurrences occurring even after doses over 70 Gy. In low grade tumours radiation does not seem to alter the overall pattern of disease progression significantly. Therefore despite use of the new technologies now available that allow radiotherapy to be delivered more accurately and to higher doses, local control of these tumours is still rarely achieved. Unfortunately these tumours have not proven sensitive to changes in radiotherapy fractionation or to the addition of radiosensitising agents. Novel approaches to these tumours are needed, based on an improved understanding of both tumour and normal tissue response to radiation. PMID- 14531560 TI - Interstitial stereotactic radiosurgery. AB - Stereotactic radiosurgery by means of interstitial application of either radionuclides or radiation devices has been used extensively in primary and secondary brain tumors. A few centers have gained sufficient expertise and clinical data to scientifically evaluate this treatment modality. Interstitial stereotactic radiosurgery is limited to circumscribed lesions with a diameter of 3.5 cm or less. The radiobiology of interstitial radiosurgery is quite well elaborated as to doses, dose rates and effects on vascular physiology. Efficacy in low grade gliomas is well documented by several European centers using 125-J sources. Different modes of implantation have been used and evaluated including single fraction treatment using a miniature linear accelerator (Photoelectron). In malignant gliomas interstitial radiosurgery has been investigated in a prospective, randomised, controlled trial and not shown to be effective. Steretactic interstitial radiosurgery is a powerful treatment option in circumscribed CNS tumors like some low grade gliomas and metastasis but does play no major role in the treatment of malignant glioma. PMID- 14531562 TI - Non-invasive transcranial high intensity focused ultrasound (HIFUS) under MRI thermometry and guidance in the treatment of brain lesions. AB - Non-invasive transcranial high intensity focused ultrasound (HIFUS) therapy given under MRI thermometry and image guidance to awake patients lying within the bore of a 1.5 T MRI scanner (a) to thermally ablate brain lesions such as metastases, (b) to cause precise ablative brain lesions in functional disorders, or (c) to locally open the blood-brain-barrier for targeted therapeutic construct delivery- without the radiation risks of stereotactic radiotherapy--may sound science fiction. Kullervo Hynynen, a Finnish-born ultrasound and MRI physicist, and Ferenc Jolesz, a Hungarian-born neurosurgeon and visionary of image guided surgery, have joined forces at Radiology, Brigham & Women's Hospital, Boston, and they have taken every step to realize the vision above, in highly successful collaboration with the industry (GE, InSightec, TxSonics). The sophisticated transcranial HIFUS instrumentation, supported by profound research data from experimental animals and by the clinical experience from extracranial HIFUS targets (breast fibroadenoma, uterine fibroid), is now coming to a phase I clinical trial in cerebral metastases. It remains to be seen whether transcranial HIFUS will find applications in diffuse gliomas such as (a) thermal ablation of selected areas of glioma tissue, (b) opening the blood-brain-barrier for therapeutic constructs to enter selected areas, or (c) activating such constructs in desired areas. The prophecy of Dr. Jolesz, "this technology will put neurosurgeons out of business", may not fulfill during our lifetime. PMID- 14531561 TI - Implantable drug-releasing biodegradable microspheres for local treatment of brain glioma. AB - Our group have developed a new method of drug delivery into the brain using implantable biodegradable microspheres. In this mini-review, we describe the development, preclinical studies and clinical trials involving 5-fluorouracil releasing microspheres for interstitial radiosensitization of malignant glioma. Future developments concerning these microspheres for treatment of brain tumors are presented. PMID- 14531563 TI - Intracavitary chemotherapy for glioblastoma: present status and future directions. AB - Considerable efforts have been invested to improve local control of the glioma disease although its infiltrative nature leading to whole brain involvement is a fundamental characteristic and antagonistic to this endeavour. The typically local recurrence of glioblastoma in about 80% of the cases has prompted intracavitary treatments of which presently only a biodegradable wafer containing carmustine has shown statistically significant benefit regarding survival in three phase III trials. Based on that proof of principle, many new developments are attempting to improve on this concept, introducing different agents with otherwise high systemic toxicity and poor penetration. New pharmacological formulations offer longer sustained release, better adaptation to the geometry of the resection cavity, and allow repeated administration. Should local recurrence become effectively controlled, significant progress can be made to increase survival with very limited local and virtually no systemic side effects. Since all agents so far show only limited activity against solid tumor, complete resection seems to be the prerequisite for effective local therapies. PMID- 14531564 TI - Intralesional radioimmunotherapy in the treatment of malignant glioma: clinical and experimental findings. AB - In the last two decades radioimmunotherapy has been used as an additional treatment option for malignant glioma in several centers. More than 400 patients have been reported, who were treated in the framework of different studies. Most of them received labelled antibodies to tenascin, an extracellular matrix glycoprotein, which is expressed in high amounts in malignant gliomas. We report side effects and survival time of 46 patients, treated after surgical resection and conventional radiotherapy with intralesionally injected labelled (131-Iodine) antibodies to tenascin. Despite the fact, that many treatments have been performed, little is known about the distribution properties of labelled antibodies after injection in the tumour cavity. For an optimal effect labelled antibodies should be able to reach tumour cells, which have migrated into the surrounding tissue. We investigated the propagation velocity and area of distribution of labelled antibodies and their considerably smaller fragments after the injection in C6-gliomas of Wistar rats. Propagation increased with time and was significantly greater after injection of labelled fragments than after injection of labelled antibodies. According to our results labelled fragments might be better able to reach distant tumour cells in the peritumoural tissue of malignant gliomas than labelled antibodies. PMID- 14531565 TI - Intracavitary treatment of malignant gliomas: radioimmunotherapy targeting fibronectin. PMID- 14531566 TI - Comparing monoclonal antibodies and small peptidic hormones for local targeting of malignant gliomas. AB - Monoclonal antibodies, F(ab')2 fragments and peptidic vectors have been clinically tested for systemic and locoregional treatment of malignant gliomas. Since these brain-intrinsic neoplasms are characterized by relentless tumor cell infiltration of normal brain parenchyma, targeting agents require diffusive properties in order to reach invading tumor cell clusters that migrate along vascular clefts and axonal pathways. Tumor uptake was significantly improved by using small peptidic hormone receptors, e.g. modified octreotide, following systemic injections as compared to macromolecules which only led to limited stabilization of the disease. More importantly, biodistribution was found to be superior following direct intratumoral injection by using these small drug-like radioconjugates. Rapid and extensive distribution within 30 minutes was observed in large tumors, even crossing the corpus callosum in bihemispheric lesions following injection of 2-3 ml of the radiopharmakon injected into the center of non-resected tumors. Distribution was far more extensive after direct intratumoral injection as compared to intracavitary injection after surgical debulking. Increased interstitial pressure gradients and the much larger and chaotic structure of the interstitial space of a tumor compared to the extremely tight architecture of normal brain tissue might explain this unexpected biodistribution pattern. Peptidic hormone vectors might become useful agents to deliver radiopharmaceuticals into human invasive gliomas. PMID- 14531567 TI - Local convection enhanced delivery of IL4-Pseudomonas exotoxin (NBI-3001) for treatment of patients with recurrent malignant glioma. AB - PURPOSE: This was an open-label, dose-escalation trial of intratumoral administration of IL-4 Pseudomonas Exotoxin (NBI-3001) in patients with recurrent malignant glioma. PATIENTS AND METHODS: A total of 31 patients with histologically verified supratentorial grade 3 and 4 astrocytoma were studied. Of these, twenty-five patients were diagnosed with glioblastoma multiforme (GBM) while six were diagnosed with anaplastic astrocytoma (AA). Patients were over 18 years of age and had Karnofsky performance scores > or = 60. Patients were assigned to one of four dose groups in a dose-escalation fashion: 6 microg/ml x 40 ml, 9 microg/ml x 40 ml, 15 microg/ml x 40 ml, or 9 microg/ml x 100 ml of NBI 3001 administered intratumorally via stereotactically placed catheters. Patients were followed with serial MRI scans and clinical assessments every four weeks for the first 16 weeks and then every eight weeks until week 26. RESULTS: No drug related systemic toxicity, as evident by lack of hematological or serum chemical changes, was apparent in any patients; treatment-related adverse effects were limited to the central nervous system. No deaths were attributable to treatment. Drug-related Grade 3 or 4 toxicity was seen in 39% of patients in all dose groups and 22% of patients at the maximum tolerated dose of 6 microg/ml x 40 ml. The overall median survival was 8.2 months with a median survival of 5.8 months for the GBM patients. Six-month survival was 52% and 48%, respectively. Gadolinium enhanced magnetic resonance imaging of the brain showed areas of decreased signal intensity within the tumor consistent with tumor necrosis following treatment in many patients. CONCLUSIONS: NBI-3001 appears to have an acceptable safety and toxicity profile when administered intratumorally in patients with recurrent malignant glioma. PMID- 14531568 TI - Convection enhanced delivery of IL13-PE38QQR for treatment of recurrent malignant glioma: presentation of interim findings from ongoing phase 1 studies. AB - IL13PE38QQR is a recombinant toxin composed of the enzymatically-active portion of Pseudomonas Exotoxin A conjugated with human IL13. Binding of IL13-PE38 to the IL13 receptor (IL13R) permits internalization of the recombinant toxin resulting in selective and potent cytoxicity at nanomolar concentrations. Normal brain tissue expresses little or no IL13R, but malignant gliomas overexpress IL13R conferring the selective cytotoxicity to the agent. Convection-enhanced delivery (CED), a novel direct drug delivery method to tumor and peritumoral region uses positive pressure infusion to generate a pressure gradient that optimizes distribution of macromolecules within the brain. Three phase I studies have been initiated to investigate IL13-PE38QQR as an anti-tumor agent for the treatment of patients with recurrent malignant gliomas. As of January 2003 a total of 46 patients have been treated. The presentation at the March 2003 EANS Local Therapy of Glioma meeting reflects adverse event findings through January 2003 and survival data through March 2003. Intratumoral infusion with or without resection is fairly well-tolerated with corticosteroids prophylaxis particularly for patients with raised intracranial pressure. Post-resection infusion into the peritumoral brain parenchyma also appears to be very well tolerated. Histopathological tumor effect was seen at drug concentrations of 0.5-2.0 microg/mL. Although phase I studies do not focus on efficacy evaluation, prolonged survival times have been observed in this select population of patients. The preclinical data and details and preliminary results of the three clinical trials are reviewed. PMID- 14531569 TI - Oncolytic viruses for treatment of malignant brain tumours. AB - Wild type viruses have been known for decades for their capability to destroy malignant tumour cells upon infection and intracellular replication. Genetic engineering of such viruses was, however, only recently done in an attempt to improve their utility as biological anticancer agents. Wild type or recombinant viruses able to selectively destroy tumour cells while sparing normal tissue are known as oncolytic viruses. Most oncolytic viruses currently investigated in clinical trials are derived from adenovirus (AV) or herpes simplex virus type I (HSVI). More than 300 patients with solid tumours were now treated in clinical trials with oncolytic viruses, and in most cases virus was administered directly into the tumour mass. About 10% of the above patients had recurrent malignant glioma. Total intratumoral doses of up to 2 x 10(12) virus particles were well tolerated, and in general no severe side effects resulted from the clinical use of oncolytic AV and HSVI, either in the brain or in the rest of the body. Encouraging anti-tumoral activity was demonstrated in some types of tumours treated locally with oncolytic viruses, and systemic chemotherapy was found to potentiate the anti-tumour effect of virus mediated oncolysis. In malignant glioma, standard gene therapy approaches employing non-replicating virus vectors failed to demonstrate significant benefit in clinical studies. Therapy with oncolytic viruses seems to hold more promise in early clinical trials than gene therapy with non-replicating virus vectors. However, further major advancements in virus designs, application modalities, and understanding of the interactions of the host's immune system with the virus are clearly needed before oncolytic virus therapy of malignant brain tumours can be introduced to clinical practice. PMID- 14531570 TI - Gene therapy of gliomas. AB - Malignant glioma formation is associated with characteristic genetic alterations, although epigenetic mechanisms may contribute in tumorigenesis. Until recently, our knowledge has mainly been based on chromosomal and molecular studies performed in the last two decades. This has increased tremendously with the advent of new technologies, in particular expression arrays for simultaneous analysis of thousands of genes. Consequently, gene therapy of gliomas may aim at molecular interference with 'gain of function' genes (oncogenes) or replacement of 'loss of function' genes (tumor suppressor genes). Such approaches require transgene expression in whole tumor cell populations (if not other mechanisms come into play) which cannot be achieved with current vector systems. Hence other strategies have been pursued which may be independent of genes actually involved in tumorigenesis. Microbial genes (e.g. herpes simplex virus thymidine kinase) may be transferred into the tumors allowing for prodrug activation (e.g. ganciclovir). Furthermore, cytokines or other immunomodulatory genes may be used for vaccination purposes which frequently involves ex vivo transfection of autologous tumor cells with such genes. These approaches proved promising in preclinical studies performed in cell culture and different inbred rodent models. A considerable number of clinical trials have been initiated based on these approaches. Although most therapeutic strategies proved safe, clinical responses fell short of expectations raised by preclinical results. This, to a large extent, has to be attributed to a lag in the development of efficient vector systems. Although much effort has been put into this area of research, neuro oncologists are still in await of a vector system allowing for selective and efficient tumor cell transduction. This has led to increased interest in distinct but related strategies, e.g. oncolytic viruses or direct intra-tumoral delivery of anti-sense oligonucleotides. PMID- 14531571 TI - Cell therapy using encapsulated cells producing endostatin. AB - Despite aggressive surgery and post-operative radiation and chemotherapy, the prognosis is poor for glioblastoma patients. Anti-angiogenic therapy with compounds such as endostatin could delay the onset of relapse. However, the short systemic half-life of this proteins as well as the blood-brain barrier makes the use of this therapy difficult for brain cancer patients. The aim of this project is to develop and implant genetically engineered producer cells secreting endostatin that are encapsulated in calcium cross-linked alginate gel beads. Encapsulation of cells within alginate gels has a potential as a sustained release system in addition to the fact that the encapsulation technology protects the cells from rejection by the immune system. Human embryonal kidney 293 cells have been transfected with the gene for endostatin. These cells have been encapsulated in calcium cross-linked alginate gels and optimized for the secretion of endostatin. Alginate gel beads implanted into rat brain have shown only a moderate loss in cell viability but extended endostatin release for periods of up to 12 months. Visualization of the anti-angiogenic effect on C6 rat glioma growth, tumor vasculature and microhemodynamics has been demonstrated by using intravital video microscopy. The data indicates that endostatin greatly affects tumor-associated microcirculation but does not appear to affect normal microcirculation. The local delivery of endostatin seems to specifically affect tumor-associated microvessels by reduction of the vessel density, diameter and functionality. Tumor cell migration and invasion was greatly reduced in the endostatin treated animals. PMID- 14531572 TI - Stem cells in neuro-oncology--development, regeneration and treatment. PMID- 14531573 TI - Glioma invasion--pattern of dissemination by mechanisms of invasion and surgical intervention, pattern of gene expression and its regulatory control by tumorsuppressor p53 and proto-oncogene ETS-1. AB - The capacity of glioma cells to invade normal brain leads to far reaching dissemination of these tumors limiting surgical resection and the prospect of local treatment strategies. The analysis of the dissemination pattern, the molecular substrates and mechanisms involved suggest that glioma invasion most likely represents independent cellular behaviors leading to distinct pattern of spread. The search for common denominators of the invasive phenotype has demonstrated that invasive cells show gene expression profiles indicating elevated expression of motility associated genes and genes involved in resistance to apoptosis whereas proliferation and apoptosis related gene expression is repressed. Confirming these findings, invasive cells in vitro show elevated motility and resistance to drug induced apoptosis. Obviously invasiveness is an early event in the progression of glial tumors. Therefore, the loss of control over invasion genes must involve key elements of molecular tumor progression. We have recently demonstrated that deregulation of invasion gene expression occurs as a consequence of functional impairment of tumor suppressor p53 leading to unrestrained activation of proto-oncogene Ets-1 dependent invasion-associated genes. In this article the prospect of limiting the dissemination of glial tumors by anti-invasive strategies and targets for modulation of the invasive phenotype to reconstitute chemo- and radiation sensitivity are discussed. PMID- 14531574 TI - Mechanisms of glioma cell invasion. AB - Invasive growth is one of the characteristics of gliomas--local infiltration into the surrounding nerve tissue decisively restricts all treatment strategies. Particularly the merit of all local treatment modalities is queried. The question whether a glioma represents a diffuse disease of the CNS or a local disturbance with unrestrained expansion tendency is still at issue. Understanding of the invasion mechanisms is of importance inasmuch as biologically reasonable and effective strategies of limiting and suppressing glioma invasion can only hence be derived. The affinity of glioma cells towards certain structures of the extracellular matrix as well as taking advantage of tumour vascularisation with regard to extension play a decisive role. Still not fully understood are tumour host interactions. Future thinking will have to take into account these interactions as well as evidence to be derived from development neurobiology and regeneration capacity of the CNS. The present review is meant to give a short overview and disclose many questions. PMID- 14531575 TI - Invasion as limitation to anti-angiogenic glioma therapy. AB - The inhibition of tumor angiogenesis could be an efficient therapeutic strategy for the treatment of malignant gliomas. Prominent neovascularization is induced by these tumors, and microvascular proliferation is a malignancy grading criterion. However, glioma cells can also invade the brain diffusely over long distances without necessarily requiring angiogenesis. Experimentally, it was shown that especially during early stages of growth in rodent brain, glioma cells can coopt the preexistent host vasculature to recruit their blood supply in the absence of neovascularization. This phenomenon was only observed in orthotopic models in which the tumor cells were implanted into the brain which is a densely vascularized environment, but not in subcutaneous models in which tumor cells are implanted into a virtual space. Using an orthotopic mouse model, we analyzed whether systemic anti-angiogenic therapy with an antibody against the vascular endothelial growth factor receptor-2 (VEGFR-2) could inhibit intracerebral growth of xenografted human glioblastoma cells and what effect this treatment had on tumor morphology and invasiveness. We found that anti-angiogenic therapy inhibited tumor growth by 80% compared to buffer-treated controls. The intratumoral microvessel density was reduced by at least 40% in treated animals compared to controls. However, in mice treated with the anti-VEGFR-2 antibody, we noticed a striking increase in the number and total area of small satellite tumors clustered around the primary mass. These satellites usually contained central vessel cores, and tumor cells often had migrated along blood vessels over long distances to eventually reach the surface and spread in the subarachnoid space. Systemic anti-angiogenic therapy can thus apparently increase the invasiveness of gliomas in the orthotopic model. Tumor cell invasion was tightly associated with preexistent blood vessels, suggesting that increased cooption of the host vasculature could represent a compensatory mechanism that is selected for by inhibiting adequate tumor vascularization. PMID- 14531576 TI - Commentary: Nature-nurture interplay in emotional disorders. PMID- 14531577 TI - A twin study of anxiety-related behaviours in pre-school children. AB - BACKGROUND: From middle childhood onwards, substantial evidence points to phenotypic differentiation between anxiety diagnostic categories such as generalised anxiety, separation anxiety, specific phobia, and obsessive compulsive disorders. However, little is known about the genetics of these categories and especially about the phenotypic and genetic structure of related behaviours in pre-school children. METHODS: We examined the phenotypic differentiation and genetics of mother-reported anxiety-related behaviours in 4,564 four-year-old twin pairs, from a population-based sample. RESULTS: Confirmatory factor analyses provided support for five correlated factors: General Distress, Separation Anxiety, Fears, Obsessive-Compulsive Behaviours, and Shyness/Inhibition. Genetic influences were found on all five factors, but the pattern of influences differed considerably across them, with particularly high heritability estimates for Obsessive-Compulsive Behaviours and Shyness/Inhibition, and substantial shared environmental influence on Separation Anxiety. Multivariate genetic analyses revealed moderate genetic correlations between the five factors. Genetic overlap was particularly pronounced between General Distress and the other anxiety-related behaviours, accounting for about half of their covariance. Genetic variance on Obsessive-Compulsive Behaviours was the least correlated with the other scales. The shared environmental influences correlated highly across the factors, accounting for the greatest proportion of covariation between Separation Anxiety, Fears and Obsessive-Compulsive Behaviours. The non-shared environment influences were largely variable specific. CONCLUSIONS: These data provide evidence for phenotypic and genetic overlap as well as differentiation between aspects of anxiety-related behaviours in young children. We conclude that research with young children will benefit from more specific assessments of anxiety-related behaviours in addition to less differentiated assessments of 'internalising' symptoms. PMID- 14531579 TI - Depressive symptoms in children and adolescents: changing aetiological influences with development. AB - BACKGROUND: Evidence suggests that depressive symptoms become increasingly heritable as children grow into adolescence. However, the literature is not entirely consistent in this respect and existing longitudinal twin studies have examined changes within adolescence only. METHOD: Parent and self-report questionnaire data were used to examine the genetic and environmental influences on depressive symptoms in a UK sample of 670 twin pairs aged 5-17. Age effects were examined cross-sectionally and longitudinally using data collected over a 3 year period. RESULTS: Cross-sectional analyses showed that shared environmental effects had significant influence in younger children but not in adolescence, when depression scores were significantly more heritable. The results of these cross-sectional analyses were supported when two waves of parent-report data collected over three years were analysed. Significant new genetic influences emerged in adolescence but no new shared environmental influences. Some sex differences were found, with girls showing greater genetic influence than boys, but only from parent-report data. CONCLUSIONS: These findings support and extend earlier work which has shown increasing genetic influence on depressive symptoms as children grow into adolescence. PMID- 14531578 TI - Sibling-based association analyses of the serotonin transporter polymorphism and internalizing behavior problems in children. AB - BACKGROUND: Childhood internalizing problems are often precursors in the development of more serious psychiatric syndromes including anxiety and depressive disorders. Twin studies of the etiology of these disorders suggest that the genetic risk factors underlying anxiety and depression are highly correlated. However, the specific genetic mechanisms responsible for this risk have not yet been identified. METHODS: We examined the association between childhood internalizing problems and a functional polymorphism in the serotonin transporter gene (5-HTTLPR) in 711 children participating in a longitudinal twin study of behavioral and emotional development. Internalizing problems were measured at ages 4, 7, 9, 10, 11 and 12 years using the Child Behavior Checklist (CBCL) parent report form. We applied a sibling-based methodology for estimating allelic association with quantitative traits, while controlling for population stratification. RESULTS: No associations were found for CBCL Internalizing problems at any age, including the subscales for Somatic Complaints, Withdrawn and Anxiety/Depression. CONCLUSIONS: Thus, although our results did not support the hypothesis that the 5-HTTLPR contributes to a dimensional expression of internalizing behavior problems, this does not rule out the possibility that it is an interesting polymorphism to pursue in the search for genetic risk factors related to major depressive and/or anxiety disorders. PMID- 14531580 TI - Negative life events as an account of age-related differences in the genetic aetiology of depression in childhood and adolescence. AB - BACKGROUND: Many twin studies have reported that the genetic aetiology of depression differs according to age, with genetic influences being more important for adolescents than younger children. We sought to examine whether this age related increase in the relative importance of genetic factors is due to an increase in gene-environment correlation specifically involving negative life events. METHOD: Questionnaires were sent to the families of a population-based sample of twins aged between 8 and 17 years. Parents of all the twins and adolescents aged 11 and over were asked to complete the Mood and Feelings Questionnaire (Costello & Angold, 1988) and a modified version of the Life Events Checklist (Johnson & McCutcheon, 1980). Responses were obtained from 1468 families and data were analysed using genetic model fitting. Bivariate analysis of 1) negative life events and 2) behaviour-dependent life events and depression symptoms was undertaken separately for children (aged 8 to 10 years) and adolescents (aged 11 to 17 years). Differences in genetic and environmental parameters across age groups were tested. RESULTS: Adolescence was associated with a greater number of behaviour-dependent life events. Genetic covariation of negative life events and depression was greater for adolescents than for children. CONCLUSIONS: Bivariate model fitting was consistent with the greater heritability of depression seen in adolescence being due to an increase in gene environment correlation involving negative life events. However, the effects of genes associated specifically with maturation in adolescence, the possibility of 'person' effects and the role of other environmental factors also need to be considered. PMID- 14531581 TI - Genetic epidemiology of self-reported lifetime DSM-IV major depressive disorder in a population-based twin sample of female adolescents. AB - BACKGROUND: In adults, about 40% of the variance in risk of Major Depressive Disorder (MDD) is due to genetic factors, but little data exist on the heritability of youth MDD. The goal of this study was the genetic analysis of MDD in an epidemiologically and genetically representative sample of adolescent female twins. METHODS: A sample of 3416 female adolescent twins systematically ascertained from birth records was assessed using a structured telephone interview that included a comprehensive DSM-IV-based section for the diagnostic assessment of MDD. Mean subject age at time of assessment was 15.5 and participation rate exceeded 85%. Genetic modeling was conducted taking into consideration the problem of censoring, i.e., that younger adolescents were not through their period of risk for adolescent onset of MDD. RESULTS: Lifetime self reported MDD prevalence ranged from 1% under age 12 to 17.4% at age 19 and older. The genetic variance in risk of MDD was 40.4% (95% confidence interval (CI): 23.9 55.1), with the remaining variance explained by non-shared environmental effects 59.6% (95%CI: 44.9-76.1). Shared environmental effects were not significant. A significant recall bias was observed with older respondents on average reporting later onsets for their first episode of MDD. CONCLUSIONS: The genetic and environmental contributions to risk of MDD in this representative sample of female adolescent twins are remarkably analogous to findings from adult samples. These results are congruent with a conceptualization of adolescent MDD and adult MDD as having very similar etiologic determinants. PMID- 14531582 TI - Adolescent twins discordant for major depressive disorder: shared familial liability to externalizing and other internalizing disorders. AB - BACKGROUND: In adolescents, as in adults, there is often comorbidity between major depressive disorder (MDD) and many other disorders. In this study, the discordant twin method was used to investigate whether this comorbidity in adolescents may be due to a shared familial liability between MDD and other internalizing disorders, and between MDD and externalizing disorders. METHODS: We examined prevalence rates of anxiety disorders, substance use disorders, and childhood externalizing disorders in 624 seventeen-year-old same-sex monozygotic and dizygotic twin pairs. Overall prevalence of MDD in this sample was 12.9%. Twenty-seven twin pairs were concordant for MDD, 107 were discordant for MDD, and 490 were concordant for no MDD. RESULTS: Prevalence rates for disorders other than MDD were elevated in the depressed twins relative to the control twins from pairs concordant for no MDD. Prevalence rates were also elevated in the nondepressed co-twins of depressed twins. Twin concordance for disorders other than MDD was higher in twin pairs where at least one twin was depressed than in pairs where neither twin was depressed. CONCLUSIONS: These results support a shared familial liability between MDD and other internalizing disorders and between MDD and externalizing disorders in adolescents. PMID- 14531583 TI - Resolving multiple epigenetic pathways to adolescent depression. AB - BACKGROUND: Genotype x environment interaction (G x E) arises when genes influence sensitivity to the environment. G x E is easily recognized in experimental organisms that permit randomization of genotypes over fixed environmental treatments. Genotype-environment correlation (rGE) arises when genetic effects create or evoke exposure to environmental differences. Simultaneous analysis of G x E and such 'active' or 'evocative' rGE in humans is intractable with linear structural models widely used in behavioral genetics because environments are random effects often correlated with genotype. The causes of the environmental variation, therefore, need to be modeled at the same time as the primary outcome. METHODS: A Markov Chain Monte Carlo approach is used to resolve three distinct pathways involving genes and life events affecting the development of post-pubertal depression in female twins and its relationship to pre-pubertal anxiety: 1) the main of genes and environment; 2) the interaction of genes and environment (G x E); and 3) genotype-environment correlation (rGE). RESULTS: A model including G x E and rGE in addition to the main effects of genes and environment yields significant estimates of the parameters reflecting G x E and rGE. Omission of either G x E or rGE leads to overestimation of the effects of the measured environment and the unique random environment within families. CONCLUSIONS: 1) Genetic differences in anxiety create later genetic differences in depression; 2) genes that affect early anxiety increase sensitivity (G x E) to adverse life events; 3) genes that increase risk to early anxiety increase exposure to depressogenic environmental influences (rGE). Additional genetic effects, specific to depression, further increase sensitivity to adversity. Failure to take into account the effects of G x E and rGE will lead to misunderstanding how genes and environment affect complex behavior. PMID- 14531584 TI - Developmental differences in neuronal engagement during implicit encoding of emotional faces: an event-related fMRI study. AB - BACKGROUND: Prior studies document strong interactions between emotional and mnemonic processes. These interactions have been shown to vary across development and psychopathology, particularly mood and anxiety disorders. METHODS: The present study used functional neuroimaging to assess the degree to which adolescents and adults differ in patterns of neuronal engagement during implicit encoding of affective stimuli. Subjects underwent rapid event-related fMRI while viewing faces with angry, fearful, happy, and neutral expressions. A surprise post-scan memory test was administered. RESULTS: Consistent with previous findings, both adolescents and adults displayed engagement of left ventrolateral prefrontal cortex when viewing subsequently recognized stimuli. Age differences emerged in patterns of neuronal activation associated with subsequent recognition of specific face-emotion types. Relative to adults, adolescents displayed more activity in the anterior cingulate when viewing subsequently remembered angry faces, and more activity in the right temporal pole when viewing subsequently remembered fear faces. Conversely, adults displayed more activity in the subgenual anterior cingulate when viewing subsequently remembered happy faces and more activity in the right posterior hippocampus when viewing subsequently remembered neutral faces. These age-related differences emerged in the absence of differences in behavioral performance. CONCLUSIONS: These findings document developmental differences in the degree to which engagement of affective circuitry contributes to memory formation. PMID- 14531585 TI - Maternal antenatal anxiety and behavioural/emotional problems in children: a test of a programming hypothesis. AB - BACKGROUND: Previous animal investigations link antenatal stress with a range of persistent behavioural abnormalities in the offspring. The current study examined if the effect was also found in humans through middle childhood. METHODS: The current study is based on the Avon Longitudinal Study of Parents and Children (ALSPAC), a prospective, community-based study that has followed a cohort of women from pregnancy. Self-report measures of maternal anxiety and depression were assessed at repeated intervals in pregnancy and the postnatal period. Children's behavioural/emotional problems were assessed by parent report at age 47 and 81 months. Information on obstetric and psychosocial factors was obtained at several points in pregnancy and the postnatal period. RESULTS: Children whose mothers experienced high levels of anxiety in late pregnancy exhibited higher rates of behavioural/emotional problems at 81 months of age after controlling for obstetric risks, psychosocial disadvantage, and postnatal anxiety and depression (for girls, OR = 1.91, 95%CI = 1.26-2.89; for boys, OR = 2.16, 95%CI = 1.41 3.30). Furthermore, the effect at 81 months was comparable to what was previously obtained at 47 months, suggesting the kind of persistent effect proposed in the animal literature. CONCLUSIONS: There is evidence that antenatal stress/anxiety has a programming effect on the fetus which lasts at least until middle childhood. PMID- 14531586 TI - Mother-child joint activity and behaviour problems of pre-school children. AB - BACKGROUND: Behaviour problems are common among pre-school children, and a substantial proportion persist, causing significant burden to the family, schools and health services. Relatively little research has addressed the effects of positive parenting on behaviour disorder in pre-school children, particularly in larger population-based studies. METHOD: A cross-sectional postal survey of a representative, population-based sample of 800 mothers of 3 1/2-year-old children living in an outer London Borough was carried out to assess the association between mother-child joint activity and behaviour problems of pre-school children. The response rate was 70%. RESULTS: Lower levels of mother-child joint activity remained independently associated with behaviour problems of pre-school children both on a binary and a continuous scale after adjusting for a wide range of household, maternal and child circumstances. The association between low levels of mother-child joint activities and behaviour problems of the children was stronger in the presence of social problems in the family. CONCLUSION: Possible causal pathways and directions for future research and intervention are discussed. PMID- 14531588 TI - Suicidal ideations and attempts in juvenile delinquents. AB - BACKGROUND: Suicidality among adolescents is a common focus of clinical attention. In spite of links to disruptive behaviors and other types of psychopathology, it is not clear whether other factors commonly associated with suicide, such as personality and parenting, predict suicidality over and above psychopathology. The purpose of the present study was to assess suicidal ideations and attempts and their relationship to psychopathology, violence exposure, personality traits and parental rearing in Russian male juvenile delinquents with conduct disorder (CD). METHOD: Suicidality and psychopathology were assessed using a semi-structured psychiatric interview in 271 incarcerated male juvenile delinquents diagnosed with CD. Violence exposure, personality characteristics and perceived parental rearing were assessed via self-reports. RESULTS: Thirty-four percent of those diagnosed with CD (92 subjects) reported a lifetime history of either suicidal thoughts or attempts. Suicidal ideators and attempters did not differ significantly on any variable of interest, but both reported significantly higher rates of psychopathology and violence exposure than the non-suicidal group, as well as higher levels of harm avoidance, lower self directedness, and higher rates of perceived negative parental rearing. Finally, even when controlling for the relationship with psychopathology, personality and perceived parental rearing factors showed significant associations with suicidality. CONCLUSIONS: Juvenile delinquents with CD have high rates of suicidal ideations and attempts, related to a wide spectrum of psychopathology and specific personality traits. These findings suggest that a combination of intrinsic and extrinsic factors create vulnerability to stressors, which under the influence of situational factors (e.g., repeated traumatization) may lead to suicidal thoughts and acts. Factors potentially contributing to vulnerability for suicidality should be identified when planning prevention and rehabilitation efforts for troubled youth. PMID- 14531587 TI - Low- and high-level controlled processing in executive motor control tasks in 5-6 year-old children at risk of ADHD. AB - BACKGROUND: The scant research on the characteristics of Attention Deficit/Hyperactivity Disorder (ADHD) in kindergarten years curtails progress on early assessment of ADHD. METHOD: By screening a general population sample of 1317 five- to six-year-old children, four groups of children were selected. The performance of 30 children later diagnosed with ADHD was compared with 74 children later diagnosed with 'borderline ADHD' (children exhibiting all ADHD symptoms but without disruptions on two situations), 113 children later diagnosed with other psychopathology, and 126 healthy controls on computerised motor control tasks involving low- and high-level controlled processing. In addition, motor control was compared with movement speed. RESULTS: The children at risk of ADHD were in general less accurate and more variable in their movements than the children with other psychopathology and healthy controls. Under conditions of high-level controlled processing, the children at risk of ADHD were disproportionately more inaccurate and had a more unstable performance with their preferred hand than the other children. In addition, linear effects were found, with the children at risk of ADHD having the worst performance, followed by the children with 'borderline ADHD', and then both groups of control children. No significant group differences were found in movement speed. CONCLUSIONS: The main findings are interpreted as evidence for a specific deficit in high-level controlled processing in young children at risk of ADHD, now found in a motor task, rather than a response task. Furthermore, the results support the notion that ADHD represents a dimensional trait. In addition, problems in movement control (the need to allocate attentional capacity) rather than problems in movement speed distinguish children at risk of ADHD from other children. The findings are interpreted as evidence that higher-order executive processes, such as self-control and self-regulation, are already affected early in the development of ADHD. PMID- 14531589 TI - Aiding diagnosis of attention-deficit/hyperactivity disorder and its subtypes: discriminant function analysis of event-related potential data. AB - BACKGROUND: Robust evidence exists that the brain functioning of children and adolescents with attention-deficit/hyperactivity disorder (AD/HD) differs from that of normal controls. The purpose of this study was to further investigate whether event-related potential (ERP) measures of brain function could be used to reliably classify normal controls versus children and adolescents with two subtypes of AD/HD. METHODS: Behavioural and topographic ERP data from children aged 8-12 years (n = 75) and adolescents aged 13-18 years (n = 75) were entered into stepwise discriminant function analyses separating controls and subjects with AD/HD, and also subjects with AD/HD-Predominantly Inattentive type (AD/HDin) from those with AD/HD-Combined type (AD/HDcom). RESULTS: For children aged 8-12 years, controls and children with AD/HD could be separated with an overall classification accuracy of 73.3%, and the AD/HD subtypes were classified with 69.4% overall accuracy. For adolescents aged 13-18 years, 58.7% of the control vs. AD/HD subjects were correctly classified, and 62.7% of subjects in the subtypes. Classification function coefficients for each of these discriminations are presented for the purposes of cross-validation. CONCLUSIONS: In children aged 8-12 years, analysis of ERP data may aid a clinician in diagnosing AD/HD, although the clinical utility of ERP analysis is reduced for adolescents aged 13 18 years. Suggestions are made to combine ERP measures with other measures of brain function in order to improve classification accuracy, and also to predict drug response in children diagnosed with AD/HD. PMID- 14531590 TI - Two-year clinical performance of Class V resin-modified glass-lonomer and resin composite restorations. AB - While a one-year report had been previously published, this study was undertaken to evaluate the clinical performance and appearance of a resin-modified glass ionomer and a resin composite over two years. Thirty-seven pairs of restorations of FujI II LC and Z 250/Single Bond were placed in caries-free cervical erosion/abfraction lesions without tooth preparation. Restorations were clinically evaluated at baseline, 6, 12, 18 and 24 months using modified Ryge/USPHS criteria. No statistically significant difference (p = 0.13) was observed in the overall performance of the materials. Retention was 96% for the resin-modified glass ionomer and 81% for the resin composite, with no additional restorations of either material lost after one year. As previously reported, retention of the Z 250 restorations at six months was below the minimum specified in the ADA Acceptance Program for Dentin and Enamel Adhesives. The resin composite restorations generally had a better appearance, with a 100% alpha rating in color match, versus 85% for the resin-modified glass ionomer. PMID- 14531591 TI - Effect of resin viscosity and enamel beveling on the clinical performance of Class V composite restorations: three-year results. AB - This study evaluated the effect of the elastic modulus and margin configuration on the clinical performance of resin-based composite restorations in Class V non carious defects. One hundred and five cervical non-carious defects on buccal surfaces of canines and premolars were included in this study. Defects were randomly divided into three Groups and restored according to the following techniques: Group 1--no enamel bevel was placed and the defect was restored with a microfilled resin-based composite (Durafill VS); Group 2--the enamel margin was beveled and the defect restored as in Group 1; Group 3-the enamel margin was beveled and the defect was restored with a flowable resin-based composite (Natural Flow). Each group comprised 35 lesions. A total-etch, one-bottle adhesive (One-Step) was used in all groups. Retention rate, pre- and post operative sensitivity, marginal discoloration and secondary caries were determined over a three-year period and the data were analyzed statistically. At six months post-insertion, the restorations placed with beveled enamel margins resulted in 100% retention regardless of the composite used compared to a 66% retention of the non-beveled margins. At two and three years, no significant difference in retention rate was found among the three groups. Post-operative sensitivity, marginal discoloration and secondary caries were not affected by enamel beveling and restorative material. Beveled enamel margins resulted in significantly better clinical retention in the first six months only. Enamel beveling and composite viscosity appeared to not significantly affect the clinical performance of Class V non-retentive composite restorations after three years. PMID- 14531592 TI - A clinical evaluation of two in-office bleaching products. AB - This half-mouth design, two-week treatment phase, combined with an 11-week evaluation double-blinded randomized clinical trial was conducted to compare two in-office bleaching products, StarBrite (35% hydrogen peroxide) with Opalescence Xtra Boost (38% hydrogen peroxide), for degree of color change of teeth, any relapse effect (darkening) associated with discontinued use and gingival irritation and tooth sensitivity associated with use. The degree of color change and relapse was evaluated by using a colorimeter, shade guide and color slide photographs. Participants self-evaluated their gingival irritation and tooth sensitivity. They recorded daily the level of gingival irritation and tooth sensitivity experienced during the first three weeks of the study. The results of this study showed no statistical difference between products during active treatment periods and any follow-up visits using the three-color evaluation methods. Color relapse began after the bleaching treatments were finished and continued until the fifth week, after which no further significant changes appeared. Also, there was no statistical difference in gingival irritation and tooth sensitivity between the products. PMID- 14531593 TI - The clinical effect of amorphous calcium phosphate (ACP) on root surface hypersensitivity. AB - Dentin hypersensitivity is a transient condition that often resolves with the natural sclerotic obturation of dentin tubules. A method of rapidly forming calcium phosphate compounds within these tubules can mimic sclerosis and lead to rapid reduction in hypersensitivity. Amorphous calcium phosphates (ACP) can be formed in situ by the sequential application of calcium and phosphate solutions. In this clinical study, 30 patients with reported dentin hypersensitivity were randomly assigned to parallel treatment or placebo groups. In the experimental treatment group, ACP was formed by topical application of a 1.5 mol/L aqueous solution of CaCl2 followed by topical application of 1.0 mol/L aqueous K3PO4. The placebo group was treated with a topical application of 1.0 mol/L aqueous solution of KCl followed by topical application of distilled water. Treatments were repeated at the 7-day and 28-day recall appointments. Response to air and tactile stimuli were measured immediately before treatment using a visual analog scale initially on day 1, then on days 7, 28 and 180. The results showed that both the experimental and placebo treatments resulted in a reduction in hypersensitivity at 180 days. However, the ACP treatment group showed a much more rapid reduction in hypersensitivity over time. The change in sensitivity was much more apparent using the air stimulus than the tactile stimulus. These results show that topical placement of ACP can rapidly reduce dentin hypersensitivity. PMID- 14531594 TI - Fracture resistance of teeth with Class II bonded amalgam and new tooth-colored restorations. AB - This study compared the cuspal fracture resistance of posterior teeth restored with four different adhesive restorations. Fifty sound, maxillary human premolars were randomly divided into a control group and four experimental groups with 10 teeth in each. Specimens in the first group were intact teeth that were tested as unprepared. The remaining four groups received mesio-oclusodistal cavity preparations and were restored with a hybrid composite (Filtek Z250), a packable composite (Filtek P60), an ormocer (Definite) and an amalgam (SDI Permite) with an amalgam bonding agent (Amalgam Bond Plus). All groups were stored in water at 37 degrees C for 15 days and thermocycled 1000 times between 5 degrees-55 degrees C. The specimens were preloaded five times in compression to 10 kg using two metal rods that contacted only the teeth on the cuspal inclines. The teeth were then loaded occlusally in an Instron Universal Testing Machine until fracture occurred. The means of force required to fracture the teeth in each of the five groups was analyzed using one-way ANOVA and Tukey Test. The difference between the mean cuspal fracture resistance of the unprepared control teeth and those restored with amalgam groups was found to be statistically significant (p < 0.05). No significant differences in resistance to cuspal fracture were found among the restoration groups, the unprepared control group and those teeth restored with hybrid composite, packable composite and ormocer groups (p > 0.05). PMID- 14531595 TI - Radiopacity of direct esthetic restorative materials. AB - This study determined the radiopacity of 21 commercially available direct esthetic restorative materials with reference to an aluminum step wedge and an equivalent thickness of enamel and dentin. A total of 168 samples measuring 6 mm in diameter and 1 mm in thickness, with eight samples of each material, were prepared from restorative materials. Enamel and dentin samples 1-mm thick were also prepared by longitudinally sectioning eight extracted human permanent molars using a microslicing machine. The optical densities of each restorative material, along with one tooth section and an aluminum step wedge were measured from radiographic images using a transmission photodensitometer. The optical density values of the specimens were used to determine the aluminum thickness equivalent values. The data were analyzed with one-way analysis of variance (ANOVA) and Duncan's multiple range tests. The results showed statistically significant differences among materials. Tetric Ceram had the greatest radiopacity value and was higher than enamel. All materials except for the microfilled resin composite Filtek A 110 had radiopacity values greater than dentin and possessed sufficient radiopacity to meet ISO 4049 standard. Significant differences were found among materials of the same composition when compared to enamel. PMID- 14531596 TI - Fiber post adhesion to resin luting cements in the restoration of endodontically treated teeth. AB - Fiber posts are widely used in the restoration of endodontically treated teeth. Scientific evidence demonstrates that the mechanical performance of teeth restored with fiber posts in combination with resin luting cements is improved with respect to metallic post restorations. The post is cemented inside the root canal using low modulus elastic polymer resins. In this study, the mechanical resistance of four different post-cement systems was assessed by means of a micro mechanical pull-out test assisted by a simulation using the finite element methodology. This in vitro test is specifically designed to accurately characterize the post-cement interface. The results show no significant difference among the adhesion properties of the various types of post-cement systems used. PMID- 14531597 TI - Effect of bur cutting patterns and dentin bonding agents on dentin permeability in a fluid flow model. AB - This study investigated the effects of bur cutting surface roughness and bonding systems on dentin permeability. A conventional straight edged bur, cross-cut serrated bur and an extensively serrated bur were utilized with two different bonding systems. Null hypothesis was that increased surface roughness does not decrease the permeability of dentin sealing after application of bonding agents. This study incorporated a fluid flow model for measuring dentin permeability. Seventy caries-free extracted molars were used and sectioned 2 mm apical to the cementoenamel junction (CEJ). The pulp tissue was removed and the chamber cleaned using 37% phosphoric acid followed by 1:1 sodium hypochlorite solution. The specimens were mounted to plexiglass plates using cyanoacrylate cement. A flow system was established through the pulp chamber and gravity pressurized phosphate buffered saline (PBS) was forced into the chamber and the "closed system" permeability measured. Burs were used to prepare flat surfaces in dentin, and after etching with phosphoric acid, the "open tubule" permeability was determined. After applying the dentin bonding agents, fluid flow from the pulp chamber into dentin was again measured. The percentage of reduction in permeability following bonding agent application was then calculated. A two-way ANOVA found a significant (p<0.05) decrease in bonded dentin permeability in specimens prepared using the cross-cut serrated and extensively serrated bur as compared to the conventional straight-edged bur. No significant differences were detected between bonding systems and no combination of surface topography or bonding agent completely sealed the surface. PMID- 14531598 TI - Effect of food-simulating liquids on the shear punch strength of composite and polyacid-modified composite restoratives. AB - This study evaluated the effects of food-simulating liquids on the shear punch strength of two composites (Tetric Ceram [TC], Vivadent; Esthet X [EX], Dentsply), a conventional (Compoglass [CG], Vivadent) and a posterior polyacid modified (Dyract Posterior [DP], Dentsply) composite. Thirty-two specimens (10-mm in diameter and 1.6-mm thick) of each material were made, randomly divided into four groups of eight and conditioned for one week as follows--Group 1 (control): air at 37 degrees C; Group 2: distilled water at 37 degrees C; Group 3: 75% ethanol-water solution at 37 degrees C and Group 4: heptane at 37 degrees C. After conditioning, the specimens were restrained within the test apparatus and subjected to shear punch strength testing using a 3.2 mm diameter punch at a crosshead speed of 0.5 mm/minute. The shear punch strength of the specimens was computed and the data was subjected to ANOVA/Scheffe's tests at significance level 0.05. The effect of food-simulating liquids on shear strength was found to be material dependent. All materials with the exception of EX were significantly weakened by ethanol solution. For DP, a significant increase in strength was observed after conditioning in water. EX was significantly stronger than TC and DY after conditioning in air, water and ethanol solution. The shear punch strength of EX and CG was significantly higher than DY after conditioning in heptane. PMID- 14531599 TI - Mechanical properties of luting cements after water storage. AB - This study determined the effect of water storage on flexural strength (FS) and compressive strength (CS) of 12 luting cements from different material classes. In addition, the influence of the curing method on the mechanical properties was investigated. The materials examined were two zinc phosphate cements (Harvard cement and Fleck's zinc cement), two glass ionomer cements (Fuji I and Ketac Cem), three resin-modified glass ionomer cements (Fuji Plus, Fuji Cem and RelyX Luting), four resin cements (RelyX ARC, Panavia F, Variolink II and Compolute) and one self-adhesive universal resin cement (RelyX Unicem). The samples were prepared and tested according to ISO specifications. Specimens for FS and CS were loaded to fracture at a constant crosshead speed of 1 mm/minute. The mechanical properties were measured after the materials were stored in distilled water at a temperature of 37.0 +/- 1.0 degrees C for 24 hours and 150 days after mixing. In a one-way ANOVA, multiple mean value comparisons using Duncan's multiple comparison tests were performed. Resin cements had the highest flexural and compressive strengths, followed by self-adhesive universal resin cement. These materials were statistically stronger than resin-modified glass ionomer cements, glass ionomer cements and zinc phosphate cements. PMID- 14531600 TI - Bonding of photo and dual-cure adhesives to root canal dentin. AB - This study evaluated the regional tensile bond strength of a dual-cure resin composite core material to root canal dentin using photo and dual-cure adhesives and different modes of polymerization. Nineteen extracted premolars were decoronated and their root canals prepared to a depth of 8 mm and a width of 1.4 mm using Para Post drills. For the microtensile bond strength (microTBS) test, 15 roots were randomly divided into five groups and their canal walls treated with a dual-cure self-etching primer (Clearfil Liner Bond 2V Primer, Kuraray Medical Co, Japan). Adhesive resin (Clearfil Liner Bond 2V Bond A) was applied to two of the groups and light cured for 20 seconds. A dual-cure adhesive resin (Clearfil Liner Bond 2V Bond A+B, Kuraray Medical Co, Japan) was applied to the remaining three groups, one of which was light cured. The post spaces of all the groups were filled with a dual-cure resin composite (DC Core) and three were light cured for 60 seconds from a coronal direction. Chemical-cure resin composite was placed on the outer surfaces of the roots, which were then stored in water for 24 hours. They were serially sliced perpendicular to the bonded interface into eight 0.6 mm thick slabs, then transversely sectioned into beams approximately 8 x 0.6 x 0.6 mm for the microTBS test. All of the failure modes were observed under SEM and analyzed using the Kruskal-Wallis Rank test. For Knoop hardness testing, four specimens were prepared in a similar manner, two were light-cured and the other two chemically-cured. The specimens were longitudinally sectioned into two pieces, and three indentations were made at 100 microm intervals from a coronal to an apical direction in the eight halves. Data were divided into two groups (coronal/apical halves of post space) and analyzed using two-way ANOVA and Scheffe's test (p<0.05). For each curing strategy, there were no significant differences in microTBS and Knoop hardness between the coronal and apical regions (p>0.05). Light exposure of both the adhesive resin and resin composite resulted in significantly higher microTBS than chemical-cure alone (p<0.05). Light exposure also significantly increased Knoop hardness at both the coronal and apical regions (p<0.05). When the bonding resin and dual-cure resin composite were chemically-cured, failures occurred cohesively within the resin. Photo initiated polymerization of the adhesive resin and dual-cure resin composite was necessary to achieve good bonding to root canal dentin, which was not dependent upon region. PMID- 14531601 TI - Effect of delayed polishing periods on interfacial gap formation of Class V restorations. AB - This in vitro study evaluated the effect of the initial polishing period through 30 minutes, 3 hours, 12 hours, 24 hours and 1 week after setting on the gap formation around Class V restorations. Three resin-modified glass ionomers, one compomer, one conventional glass ionomer and one microfilled composite were used as controls. When specimens of the two types of glass ionomers and a microfilled composite were polished immediately after the setting procedure, this study showed 100-140 gaps around the X-section of the restorations. In contrast, only 10-40 gaps around the Class V restoration were observed when the specimens were polished after 12 hours of storage. Significant differences were observed between polishing immediately and polishing after 12 hours of storage in the two glass ionomer restorative materials. The compomer did not show this pattern. No significant differences were observed among the six polishing periods of the sum number of gaps at the cavity-restoration interfaces. The tendency of Silux Plus was similar to the two types of glass ionomer materials; namely, when the specimen was polished and inspected after storage in water for 24 hours or one week, the authors observed almost 30 gaps around the restorative cavities. PMID- 14531603 TI - Effect of application technique and dentin bonding agent interaction on shear bond strength. AB - This study evaluated the interaction of five clinical application techniques and the shear bond strength of four DBAs (OptiBond FL, Clearfil SE Bond, PQ1 and Prime & Bond NT). A hybrid resin composite (Herculite XRV restorative resin) was attached to human dentin surfaces using five application techniques: Group A- adhesive spread with a 3M brush for 30 seconds, followed by compressed air 0.5 cm from the surface for one second to remove the excess adhesive. Group B--adhesive spread with a 3M brush for 30 seconds, followed by compressed air 0.5 cm from the surface for three seconds to remove the excess adhesive. Group C--adhesive spread with 3M brush for 30 seconds, excess adhesive removed with a clean brush, two strokes side by side, no compressed air. Group D--adhesive spread with a Micro applicator brush for 30 seconds followed by compressed air 0.5 cm from the surface for one second to remove the excess adhesive. Group E--adhesive spread with a Micro-applicator brush for 30 seconds, the excess adhesive removed with a clean brush, two strokes side by side and no compressed air. The specimens were stored in distilled water at 37 degrees C for 24 hours, followed by thermocycling between 5 degrees C and 55 degrees C for 1,000 cycles. The shear bond strengths were determined on a universal testing machine operating with a crosshead speed of 5 mm/minute. The fracture sites were examined by 20x stereo microscope to determine the type of failure that occurred during the debonding procedure. Bond strength data were compared with analysis of variance at a significance level of p<0.05. Post hoc comparisons of means were performed with t-tests with p-values adjusted for multiple comparisons. This in vitro study concluded that there was an interaction between the application technique and bonding agent tested. All DBAs utilized the one-second compressed air technique, which yielded the highest bond strengths. PMID- 14531602 TI - Effect of surface finishing and storage media on bi-axial flexure strength and microhardness of resin-based composite. AB - This in vitro study tested the following null hypotheses: (1) surface finishing treatments do not significantly affect the biaxial flexure strength and microhardness of resin-based composites (RBC) and (2) storage media do not significantly affect these physical properties. Discs (81 RBC and 81 UR; 3M/ESPE) were prepared using a circular polyethylene mold (2.4-mm thick x 16.7 mm in diameter) that was polymerized through a Mylar strip and divided into three surface finishing treatment groups: 1 microm aluminum oxide slurry; 15 microm diamond and a Mylar strip. Randomly selected controls for each finishing group were stored at room temperature in individual vials. Test specimens were immersed in water, stored at 37 degrees C for two days and transferred for an additional seven days to one of three aqueous storage media at 37 degrees C: coffee (pH 5.1), cola (pH 2.4) or red wine (pH 3.7). Post immersion (nine days total), the specimens were tested for biaxial flexure strength (BFS) and Vicker's microhardness (VHN). ANOVA and Tukey's HSD test were used for statistical analysis. ANOVA results indicated that surface finishing treatments had a significant effect on the biaxial flexure strength and microhardness of the RBC and the UR specimens. BFS results for RBC specimens were AL>DD>ML (p<0.0001) and VHN results were AL, DD>ML (p<0.0001). Storage in wine medium reduced the VHN of UR specimens significantly. Both alternative hypotheses were accepted. In addition, the Mylar finishing group, because of the resin-rich surface layer, yielded the lowest mean values of BFS and VHN. PMID- 14531604 TI - Marginal adaptation of dentin bonded ceramic inlays: effects of bonding systems and luting resin composites. AB - This in vitro study evaluated the marginal adaptation of bonded inlays of lucite reinforced glass ceramic (Empress) to dentin as influenced by different bonding systems and by luting resin composites (LRCs) with different curing modes. Forty eight Empress inlays etched with 5% hydrofluoric acid and treated with a silane coupling agent (Monobond-S) were bonded to two-surface Class II cavities. Two total-etch bonding systems (OptiBond FL, Nexus) and one bonding system with selective enamel etching and a self-conditioning dentin primer (ART Bond) were included in the study. ART Bond was tested with and without the pre-curing of a first layer of adhesive resin selectively applied to the cervical cavity floor (selective double-bond technique). Each bonding system was used in combination with a light-cured resin composite (Prodigy) and a dual-cured LRC (Nexus or Vita Cerec Duo Cement). Marginal integrity was evaluated before and after thermocycling (TC) in a scanning electron microscope (SEM). Dye penetration tests were performed after TC was completed. The median percentages of continuous margin in dentin ranged from 80% to 100% before TC and from 53.5% to 96.1 % after TC. After TC, the influence of the bonding system was more pronounced than that of the LRC. In combination with the LC resin composite, ART Bond with precuring was significantly higher and the Nexus bonding system had significantly lower proportions of continuous margin than all the other bonding systems investigated. Swelling of the adhesive along the gingival margins was frequently found with the Nexus bonding system and with ART Bond without pre-curing. Microleakage was detected with all bonding system/LRC combinations, with somewhat lower rates in specimens completed using the selective double-bond technique. With the exception of the Nexus bonding system, post-TC marginal integrity was not influenced by the curing mode of the LRC (LC vs DC). In conclusion, the marginal quality of dentin bonded ceramic inlays can be improved by proper selection of the bonding sys- tem/LRC combination. The results of this study indicate the use of the Nexus luting system as directed without substitution. PMID- 14531605 TI - Shear bond strength of current adhesive systems to enamel, dentin and dentin enamel junction region. AB - This study investigated the bonding of current resin adhesives to the region approximating the dentin-enamel junction (DEJ), where the etch pattern to enamel or dentin may be different. Three kinds of tooth substrates were chosen for testing: enamel, dentin and the DEJ region. A self-etching primer system (Clearfil SE Bond) and two total-etch wet bonding systems (Single Bond and One Step) were used. Each tooth region was bonded with one of the adhesive systems, and a resin composite and was subjected to a micro-shear bond test. In addition, morphological observations were performed on debonded specimens and etched surfaces using confocal laser scanning microscopy (CLSM). CLSM observations showed that the DEJ region was etched more deeply by phosphoric acid gel than enamel or dentin, suggesting that the action of acid etch seemed to be more intense on the DEJ. However, no statistically significant differences of shear bond strength values were observed between the DEJ region and enamel or dentin, or the adhesive systems used (p>0.05). Bonding to the DEJ is potentially as good as that to enamel or dentin. PMID- 14531606 TI - Surface pH and bond strength of a self-etching primer/adhesive system to intracoronal dentin after application of hydrogen peroxide bleach with sodium perborate. AB - This study compared the dentin bond strength of a self-etching primer/adhesive system with dentin surface pH with or without bleaching and observed the morphological changes in bleached dentin treated with a self-etching primer. Dentin disks were prepared from the coronal-labial region of 32 human anterior teeth. The pulpal surfaces of the dentin disks were polished with 600-grit SiC paper under running water. The dentin surfaces on all specimens were bleached with a mixture of 30% hydrogen peroxide and sodium perborate in 100% humidity at 37 degrees C for one week. The bleaching agent was then rinsed off with water for 5, 15 or 30 seconds. All specimens were stored in water at 37 degrees C. Half of the five-second rinsing specimens were stored in water for an additional week. Dentin surface pH with or without bleaching was examined using a pH-imaging microscope (SCHEM-100). A self-etching primer/adhesive system (Clearfil SE Bond) was applied to bleached or unbleached dentin according to the manufacturer's instructions. After 24-hour water storage, the bonded specimens were prepared for microtensile testing. Microtensile bond strength (microTBS) to dentin was measured using a universal-testing machine (EZ test, Shimadzu, Japan) at a crosshead speed of 1.0 mm/minute. Data were analyzed by one-way ANOVA and Scheffe's test (alpha=0.05). The pH values of the dentin surfaces of the 5 and 15 second rinsing groups were significantly higher than the control group (p<0.05), while the 30-second rinsing and one-week water storage groups had similar surface pH values to the control group (p<0.05). The microTBS of 5, 15 and 30 second rinsing specimens after bleaching were significantly lower than the control specimens (p<0.05). However, after one-week of water storage, the microTBS returned to the control group. The application of a bleaching agent increased the pH value of the dentin surface and decreased the bond strength of the self etching primer/adhesive system. One-week water storage after bleaching recovered the surface pH and the microTBS to dentin. PMID- 14531607 TI - Evaluation of active and arrested carious dentin using a pH-imaging microscope and an X-ray analytical microscope. AB - A pH-imaging microscope was tested on carious teeth to examine the acid-base characteristics of active and arrested carious dentinal lesions and compared with an assessment of mineral loss in carious affected lesions using an x-ray analytical microscope. Extracted human molars, 22 active and 83 arrested carious teeth, were sliced vertically to maximize the presence of visible carious lesions. The pH change on the dentin surface was measured with a pH-imaging microscope, and the mineral loss was measured with an x-ray analytical microscope. The pH value of intact dentin and active and arrested carious dentinal lesions showed 7.0 (n=105), 5.8 (n=22) and 6.6 (n=83), respectively (analyzed by Kruskal-Wallis). The mineral loss was distinguishable in active dentinal lesions (4.92 times as much as intact dentin), while arrested lesions showed a slight mineral loss (2.19 times as much as intact dentin). The changes in pH value and mineral loss were significantly correlated (Pearson's correlation coefficients r=-0.8024) in active carious dentin lesions and weakly correlated (r=-0.1480) in arrested carious dentin lesions. Changes in tooth substance with the progression of the active carious process were reflected in a reduction in pH value and an increase in the amount of mineral loss. PMID- 14531608 TI - Effect of home-use bleaching gels on fluoride releasing restorative materials. AB - To evaluate the effects of two home-use bleaching gels (Nite-White and Contrast PM) on the surface roughness of one high-viscosity glass ionomer cement (Fuji IX GP), four polyacid-modified resin-based composites (Dyract AP, F2000, Elan and Compoglass F), two resin-modified glass-ionomer cements (Vitremer and Fuji II LC), a microfilled fluoride-releasing resin-based composite (Tetric) and a microfilled non-fluoridated resin-based composite (Valux) were used as controls. The null hypotheses tested were: (1) the use of bleaching gels would not result in surface roughness values different from those obtained at baseline; (2) differences in the amount of carbamide peroxide present in the gels would not influence surface roughness. Thirty specimen disks (the first 10 used for controls and the balance divided randomly into two groups) were made for each material according to the manufacturers' instructions. Finishing and polishing of the test and control specimens was performed after 24 hours using a sequential series of three (medium, fine and superfine) Sof-Lex disks, keeping the restoration surface wet. For every sequence, 10 strokes were made in one direction using a low-speed handpiece. Ultrasonic cleaning of the polished specimens was performed for two minutes in distilled water to remove any surface debris. Prior to bleaching treatment, the specimens were stored in 37 degrees C deionized water for 24 hours. The specimens were exposed to the bleaching gels for a period that simulated the equivalent of 15 days (eight hours/day) under bleaching treatment. The average surface roughness (Ra) of the control and treated specimens was measured. Three separate Ra measurements along the direction of rotation of the finishing and perpendicular to the finishing direction and edge of the mold were made for each specimen surface. After treatment with bleaching gels, there was an increase in surface roughness for F2000, Dyract AP, Elan, Valux and Tetric, while Fuji IX, Fuji II LC, Vitremer and Compoglass F displayed decreased surface roughness values, suggesting that the effects of the gels appear to be material-dependent. Although Contrast PM contains 5% more carbamide peroxide as the active ingredient, the second null hypothesis was rejected for Fuji IX, Vitremer, Dyract AP, Compoglass F, Elan and Valux. PMID- 14531609 TI - Adhesive permeability affects composite coupling to dentin treated with a self etch adhesive. AB - Dentin bonded with one-step self-etch adhesives does not couple well to chemically-cured composites despite the presence of ternary catalysts in some systems. To determine whether additional factors are responsible for the poor coupling of dentin bonded with these adhesives, the authors used a one-step self adhesive in which the ternary catalyst is incorporated separately in the brush applicator for activating the adhesive. The activated adhesive was bonded to dentin or prepolymerized composites. For microtensile bond strength evaluation, a dual-cured hybrid composite was used for coupling to the bonded substrates and activated using: (1) the light-cured mode (LC), (2) delayed light-activation (DL) and (3) the chemical-cured mode (CC). In addition, the composite polymerized in the CC mode was coupled to the two substrates bonded with (4) additional experimental adhesive versions without the ternary catalyst and (5) without the ternary catalyst and tertiary amine. Silver tracer penetration into bonded interfaces was examined using TEM by replacing the dual-cured composite with a light-cured or a chemically-cured microfilled composite. In the CC mode, the composites coupled poorly to both substrates bonded with the unactivated adhesive (5). The use of activated adhesives allowed for good coupling of the composites polymerized in all curing modes to bonded composite, but only in the LC mode to bonded dentin. Water-filled channels in the form of water trees and water blisters were present in the adhesive layer under all composite-curing modes in bonded dentin. These features were completely absent in the bonded composites, showing that the source of the water was from dentin. Water blisters were also observed along the adhesive-composite interfaces in bonded dentin when the coupling composites were polymerized using the CC or DL modes. PMID- 14531610 TI - Effect of power density on shrinkage of dental resin materials. AB - This study compares volumetric changes and rates of shrinkage during different stages of polymerization of dental resin composites and compomers exposed to the same total energy by using two different combinations of power density and exposure duration. A hybrid composite and its equivalent flowable and a compomer and its equivalent flowable were exposed using a halogen curing unit set at 400 mW/cm2 for 40 seconds and 800 mW/cm2 for 20 seconds: delivering 16 J/cm2 in both cases. Volumetric changes were recorded every 0.5 seconds using a mercury dilatometer. Ten replications per test condition were performed and the data were subjected to ANOVA. Statistically significant differences in shrinkage values and rates among different power densities were determined by means of paired t-tests at a 95% confidence level. Significantly more shrinkage (p<0.05) was found for the higher filled materials, Z250 and Dyract AP, when higher power density was used. However, no significant differences were found between their flowable counterparts when exposed to various power densities. Of the four materials, only Dyract AP exhibited no significant difference in shrinkage rate when various power densities were used. All the other materials exhibited significantly higher rates (p<0.05) at the higher power density. PMID- 14531611 TI - Wear and microhardness of different resin composite materials. AB - This study determined the three-body abrasive wear resistance of two packable composites (P-60; Solitaire 2), an ion-releasing composite (Ariston AT), a hybrid composite (Tetric Ceram) and an ormocer (Admira). The study also looked at the correlation between wear resistance and hardness of the composites. Three-body wear testing was performed using an ACTA wear machine with 15 N contact force using millet seed as the third body. Wear depth (microm) was measured by profilometry after 200,000 cycles. The hardness test was performed using a digital microhardness tester (load: 500 g; dwell time: 15 seconds). The data were analyzed by using Kruskal Wallis (p < 0.05). There were statistically significant differences among the three body abrasive wear of the composites. The ranking from least to most were as follows: Filtek P-60 < Solitaire 2 < Ariston AT < Tetric Ceram < Admira. Filtek P-60 showed the highest microhardness value. No other significant differences in hardness were observed among the different resin composites (P-60 > AristonAT = Tetric Ceram = Solitaire 2 = Admira). The results of this study indicate that there are significant differences in the wear resistance of the resin composites. The correlation between hardness and wear was significant with a correlation coefficient of r:-0.91. A significant negative correlation exists between hardness and three-body wear of resin composites. PMID- 14531612 TI - Polymerization shrinkage and microleakage in Class II cavities of various resin composites. AB - This study investigated the polymerization shrinkage of restorative materials and microleakage in the proximal box of Class II restorations. Twenty caries free extracted human molars were prepared on the mesial and distal, making 40 slot cavities. Groups (n=10) were classified as: Group 1: Single bond/Filtek Z-250; Group 2: Single Bond/Filtek Flow/Filtek Z-250; Group 3: Admira Bond/Admira; Group 4: Ariston Liner/Ariston. Dentin bonding systems and resin composites were applied according to the manufacturer's instructions. The specimens were stored in distilled water for one week at 37 degrees C, thermocycled (5 degrees-55 degrees C x 2000), stained with 50% aqueous silver nitrate, immersed in a diluted developer solution (24 hours), rinsed and sectioned in two pieces with a low speed saw (Isomet). Dye penetration (Score 0-4) was determined separately at the occlusal enamel margin and cementoenamel junction. The volumetric polymerization shrinkage of materials was determined with the apparatus of Watts and Cash (Watts Cash, 1991) for 300 seconds. Polymerization shrinkage results were as follows: Filtek Flow (3.5% +/- 0.1) > Admira (2.1% +/- 0.1) = Ariston AT (2.3% +/- 0.1) > Filtek Z-250 (1.8% +/- 0.1) (One-way Anova, post hoc Bonferroni test at p = 0.05 level). No statistical difference was found at the enamel margins for any of the materials (Score 0-1). At the cemento-enamel junction, all test groups showed significantly higher microleakage when compared to the enamel margins (Kruskal Wallis test, Mann-Whitney U). Dye penetration results at dentin were as follows: Filtek Flow + Filtek Z-250 = Admira < Ariston AT = Filtek Z-250. microleakage than the ion-releasing and hybrid composites lined only with bonding agent at the cementoenamel junction in Class II cavities. PMID- 14531613 TI - Effect of different intensity light curing modes on microleakage of two resin composite restorations. AB - This in vitro study evaluated the marginal leakage of two resin composites light cured in four different polymerization modes. Standardized Class V cavities were prepared on the buccal enamel surface of 80 sound, freshly extracted inferior bovine incisors. The teeth were randomly divided into eight experimental groups: two resin composites (Z250 Charisma) and four polymerization modes (conventional 680 mW/cm2/30 seconds; soft start-380mW/cm2/10 seconds + 680 mW/cm2/20 seconds; plasma arc curing- 1480mW/cm(2)-3 seconds; combined- 380mW/cm2-10 seconds + 1480mW/cm(2)-3 seconds). All specimens were thermocycled (1,000 cycles at 5 degrees C and 55 degrees C) with a dwell time of one minute at each temperature and immersed in a 2% methylene blue solution for more than 12 hours. The specimens were then washed and prepared for spectrophotometric analysis to quantify the dye concentration. Soft start and combined polymerization modes presented better results and were statistically different from conventional and plasma arc curing polymerization methods for both resin composites. There were no statistical differences between the two resin composites. PMID- 14531614 TI - Microtensile bond strengths of an etch&rinse and self-etch adhesive to enamel and dentin as a function of surface treatment. AB - In light of the current trend towards "minimal invasive" dentistry, diverse cavity preparation techniques have been introduced as an alternative or addition to common bur instrumentation. This study investigated whether diamond sonoabrasion (SonicSys Micro, Kavo), air abrasion (Prep Start, Danville) and Er:YAG laser irradiation (Fidelis) produce surfaces at enamel/dentin that are equally receptive to bonding as traditional mid-grit diamond-bur (Komet) and 600 grit SiC-paper prepared surfaces, of which the latter two served as controls. An etch&rinse adhesive (OptiBond FL, Kerr) applied with and without prior acid etching and a self-etch adhesive (Clearfil SE, Kuraray) were employed to bond the restorative composite (Z100, 3M ESPE) to the diversely prepared enamel and dentin surfaces. The microtensile bond strength (microTBS) was determined after 24 hours of storage in water at 37 degrees C. The results indicated that the manner of preparation of enamel and dentin prior to bonding procedures significantly influenced the bonding effectiveness of both the etch&rinse and the self-etch adhesive. Using an etch&rinse adhesive, separate acid-etching of air-abraded and Er:YAG-irradiated enamel and dentin surfaces remains mandatory. Bonding to diamond-sonoabraded and air-abraded enamel and dentin was, in general, not different from bonding to conventional diamond-bur prepared surfaces, whereas, bonding to Er:YAG-irradiated enamel and dentin surfaces in general resulted in a significantly lower bonding effectiveness compared to bonding to diamond-bur prepared surfaces. PMID- 14531615 TI - The full-length phylogenetic tree from 1551 ribosomal sequences of chitinous fungi, Fungi. AB - A data set with 1551 fungal sequences of the small subunit ribosomal RNA has been analysed phylogenetically. Four animal sequences were used to root the tree. The parsimony ratchet algorithm in combination with tree fusion was used to find most parsimonious trees and the parsimony jackknifing method was used to establish support frequencies. The full-length consensus tree, of the most parsimonious trees, is published and jackknife frequencies above 50% are plotted on the consensus tree at supported nodes. Until recently attempts to find the most parsimonious trees for large data sets were impractical, given current computational limitations. The parsimony ratchet in combination with tree fusion was found to be a very efficient method of rapid parsimony analysis of this large data set. Parsimony jackknifing is a very fast and efficient method for establishing group support. The results show that the Glomeromycota are the sister group to a monophyletic Dikaryomycota. The majority of the species in the Glomeromycota/Dikaryomycota group have a symbiotic lifestyle--a possible synapomorphy for a group 'Symbiomycota'. This would suggest that symbiosis between fungi and green plants evolved prior to the colonization of land by plants and not as a result of the colonization process. The Basidiomycotina and the Ascomycotina are both supported as monophyletic. The Urediniomycetes is the sister group to the rest of the Basidiomycotina successively followed in a grade by Ustilaginomycetes, Tremellomycetes, Dacrymycetales, Ceratobasidiales and Homobasidiomycetes each supported as monophyletic except the Homobasidiomycetes which are left unsupported. The ascomycete node begins with a polytomy consisting of the Pneumocystidomycetes, Schizosaccharomycetes, unsupported group with the Taphrinomycetes and Neolectales, and finally an unnamed, monophyletic and supported group including the Saccharomycetes and Euascomycetes. Within the Euascomycetes the inoperculate euascomycetes (Inoperculata) are supported as monophyletic excluding the Orbiliomycetes which are included in an unsupported operculate, pezizalean sister group together with Helvellaceae, Morchellaceae, Tuberaceae and others. Geoglossum is the sister group to the rest of the inoperculate euascomycetes. The Sordariomycetes, Dothideomycetes, Chaetothyriomycetes and Eurotiomycetes are each highly supported as monophyletic. The Leotiomycetes and the Lecanoromycetes both appear in the consensus of the most parsimonious trees but neither taxon receives any jackknife support. PMID- 14531616 TI - Nuclei, micronuclei and appendages in tri- and tetraradiate conidia of Cornutispora and four other coelomycete genera. AB - The distribution and behaviour of nuclei in conidia of 11 coelomycete species with tri- and tetraradiate conidia and belonging to five genera has been investigated: Cornutispora (C. ciliata, C. intermedia, C. lichenicola, C. limaciformis, and C. pittii), Eriosporella (E. calami), Furcaspora (F. abieticola, F. pinicola), Suttoniella (S. eriobotryae, S. gaubae), and Tetranacrium (T. gramineum). They have been studied by the HCl-Giemsa technique using dried, preserved material including holotypes and isotypes with ages ranging from 3 to 116 yr. Conidia of Cornutispora species showed different ploidy levels, and C. limaciformis showed a very high (> 90%) frequency of stable and viable micronuclei with an unusual type of ploidy level, occurring naturally. Frequency of ploidy levels in nuclei within conidia of Cornutispora species appeared to be associated with changes in gross conidial morphology. This is the first report of micronuclei in coelomycetes. The types of appendages on arms or parts of conidia have been studied using various stains including erythrosin in ammonia and a modified Leifson's flagella staining technique. In Furcaspora species the apical and basal conidial appendages are cellular maintaining protoplasmic continuity with the arms on which they are sited. The results have been compared with those of Crucellisporium species which have tetraradiate conidia. The new species, Cornutispora intermedia, C. pittii and Furcaspora abieticola spp. nov. are described, and illustrated, and a key to all known Cornutispora species is provided. PMID- 14531617 TI - Rhodotorulic acid production by Rhodotorula mucilaginosa. AB - Rhodotorula mucilaginosa produces the siderophore rhodotorulic acid (RA) when grown in iron-limited conditions. R. mucilaginosa grew at rates between 0.10 and 0.19 h(-1) in iron-restricted conditions, depending on the carbon source, and at 0.23 h(-1) in iron-sufficient conditions. In bioreactors inoculated with iron starved pre-cultures, initial specific growth rates in batch culture were dependent on the iron concentration. The critical dilution rate (Dcrit, at which steady state cultures cannot be sustained) in continuous cultures was also dependent on the iron concentration and was lower than mu(max) in batch culture. Sucrose was the best carbon source for RA production [287+/-11 micromol (g biomass)(-1)] and production could be further increased by supplementing the medium with the precursors acetate [460+/-13 micromol (g biomass)(-1)], ornithine [376+/-6 micromol (g biomass)(-1)], or both [539+/-15 micromol (g biomass)(-1)]. Citric acid was an effective suppresser of RA production. RA was produced in a growth rate dependent manner and was optimally produced at pH 6.5. PMID- 14531618 TI - Heterobasidiomycetes form symbiotic associations with hepatics: Jungermanniales have sebacinoid mycobionts while Aneura pinguis (Metzgeriales) is associated with a Tulasnella species. AB - In order to evaluate substrate dependence of the symbiotic fungal associations in leafy liverworts (Jungermanniopsida), 28 species out of 12 families were investigated by transmission electron microscopy and molecular methods. Samples were obtained from the diverse substrates: from naked soil, from the forest floor on needle litter, from between peat moss, from rotten bark of standing trees, and from stumps and rotten wood. Associations with ascomycetes were found in most of the specimens independent from the substrate. Seven species sampled from soil were found to contain basidiomycete hyphae. Ultrastructure consistently showed dolipores with imperforate parenthesomes. Molecular phylogenetic studies revealed that three specimens belonging to the Jungermanniales were associated with members of Sebacinaceae, while Aneura pinguis (Metzgeriales) was associated with a Tulasnella species. These taxa are so far the only basidiomycetes known to be symbiotically associated with leafy liverworts. The probability that the associations with Sebacinaceae are evolutionary old, but the Tulasnella associations more derived is discussed. The sebacinoid mycobionts form a similar interaction type with the jungermannialian leafy liverworts as do the associated ascomycetes. The term 'jungermannioid mycorrhiza' is proposed for this distinctive symbiotic interaction type. PMID- 14531619 TI - Taxonomy and toxicity of Conocybe lactea and related species. AB - Conocybe lactea was examined as part of a larger study on the distribution of amatoxins and phallotoxins in fungi, and the taxonomic relationships between these fungi. As amatoxins are present in the congener C. filaris, the locally abundant C. lactea was examined using HPLC and mass spectroscopy. Amatoxins were not found in C. lactea, but the related phallotoxins were present in small quantities making it the first fungus outside of the genus Amanita in which phallotoxins have been detected. Despite the presence of a related toxin, C. lactea was found not to be taxonomically close to C. filaris. Phylogenetic analyses using nuclear ribosomal RNA genes indicated that North American specimens of C. lactea were conspecific with North American specimens of C. crispa in Conocybe sect. Candidae. European C. crispa was a distinct taxon. The implications of the use of the name C. albipes for these taxa are discussed. Nucleotide data confirmed placement of the sequestrate taxon Gastrocybe lateritia in sect. Candidae, but as a distinct taxon. It is hypothesized that the unique sequestrate morphology of G. lateritia may be caused by a bacterial infection. PMID- 14531621 TI - Confocal imaging reveals structural detail of the cell nucleus and ascospore formation in lichenized fungi. AB - A method for visualization of fungal nuclear structures using the DNA specific fluorochrome SYTOX Green and confocal laser scanning microscopy is presented. The staining procedure detects nuclei in different cell types with high contrast. Furthermore, the method permits generation of high-resolution confocal image stacks using optimised scanning conditions. This approach significantly improved visualization of stained fungal nuclear structures. The technique was applied to some lichenized fungi, but can also be used to study similar detail in non lichenized fungi. In certain cases, incompletely separated chromosome-like structures, heterochromatic material as well as weakly stained nucleolar regions can be seen. In Xanthoria parietina, variation in the nuclear division was observed in developing ascospores. Delayed division after the formation of endospore thickening resulted in the exchange of nuclear material through a narrow isthmus. PMID- 14531620 TI - Production of swainsonine by fungal endophytes of locoweed. AB - Consumption of locoweeds, legumes endemic in arid western USA, has long been associated with locoism, a disease of ruminant animals. To explore the relationship between fungi associated with locoweed and locoweed toxicity, 11 locoweed populations from various sites in New Mexico were assessed for endophytic fungi. Endophytes were isolated from the leaves, stems, seeds, and flowers of eight populations of the toxic locoweeds Astragalus mollissimus, Oxytropis lambertii, and O. sericea. Fungal cultures grew very slowly and sporadically produced subcylindrical conidia with very dark transverse septa. All cultured endophytes produced the alkaloid swainsonine, which causes locoism. Endophyte-infected locoweed populations produced swainsonine, and the swainsonine level of endophyte strains in vitro was highly correlated with the swainsonine level of their host plant populations. The rDNA ITS from mycelia from four endophyte isolates and beta-tubulin encoding regions from mycelia of 18 fungal endophyte isolates were amplified using PCR and the nucleic acid sequences were analyzed. The nucleic acid sequences of the beta-tubulin encoding regions were essentially identical among all the endophytes regardless of plant genus and locations. Morphological evidence and sequence analysis of the ITS region suggest that the endophytes are most closely related to Embellisia. However, with the paucity of Embellisia species represented in sequence databases, precise taxonomic placement will await further study. PMID- 14531622 TI - Enzyme production by Mycena galopus mycelium in artificial media and in Picea sitchensis F1 horizon needle litter. AB - Mycena galopus is among the most important leaf litter decomposers in UK coniferous and angiosperm woodlands, having the potential to utilise all the major constituents of plant litter. Even so, the enzyme or combination of enzymes produced by M. galopus responsible for lignin depolymerisation was previously unknown. A range of media from liquid and semi-solid cultures to more natural substrata was tested to determine whether laccase was produced by an isolate of M. galopus, M9053. Malt extract liquid medium (MEL) with 2,5-xylidine favoured laccase production as compared with the same medium containing the inducers veratryl alcohol, veratryl aldehyde, veratric acid, homoveratric acid, vanillic acid or p-anisic acid. A semi-solid medium of cereal bran in phosphate buffer and a solid medium of Picea sitchensis F1 horizon needle litter were also not as effective as MEL with 2,5-xylidine as an inducer. Compared with six other isolates of the same species grown in MEL without inducers, M9053 exhibited rates of laccase activity fairly typical for M. galopus. An isolate from a dark coloured basidiome of M. galopus, but not var. nigra, exhibited the greatest activity while var. candida showed relatively low laccase activity. Marasmius androsaceus exhibited peak laccase production several days later than M. galopus. In addition, a manganese-dependent peroxidase that was responsible for 15% (in MEL culture fluid) and 39% (in needle litter extract III) of ligninolytic activity was produced by M9053. A further peroxidase was found to be the major ligninolytic constituent in MEL extracts (53%), and had a similar contribution to total activity (29%) as laccase (32%) in needle litter fraction III. Mycena galopus produced water- and buffer-extractable mannases and xylanases when grown on needle litter. PMID- 14531624 TI - Fundus autofluorescence in patients with pseudoxanthoma elasticum. AB - PURPOSE: To characterize changes in fundus autofluorescence in patients with pseudoxanthoma elasticum (PXE). Fundus autofluorescence intrinsically derives from lipofuscin, and the degree of autofluorescence is thought to indicate the degree of retinal pigment epithelium (RPE) metabolic activity. METHODS: Twelve eyes of 6 patients (2 men, 4 women) with PXE were studied with a confocal scanning laser ophthalmoscope. Patient age ranged from 42 to 62 years. The autofluorescence of abnormal retinal areas was compared digitally with that of neighboring, presumed healthy control areas. When the average gray level of a fundus region was 2 SDs above or below the average gray level of a control area, autofluorescence of the fundus region was considered abnormal. RESULTS: In all 12 eyes, some segments of the angioid streaks showed decreased fundus autofluorescence, and other segments of the streaks showed normal autofluorescence. Areas of peripapillary chorioretinal atrophy seen in 2 eyes and of disciform scarring seen in 3 eyes showed decreased autofluorescence. Solitary or multiple drusen-like spots showed increased autofluorescence in all 12 eyes. CONCLUSION: Atrophic and degenerative RPE regions showed decreased fundus autofluorescence in areas of chorioretinal atrophy and in some segments of the angioid streaks. Some drusen-like spots showed increased autofluorescence. The characteristic changes in autofluorescence that we observed in PXE patients suggest that the content of the drusen-like substance differs from that of senile drusen and that the drusen-like lesions are similar to the sub-RPE deposits seen in macular dystrophy. PMID- 14531623 TI - Effects of systemic octreotide, local mytomycine-C and local corticosteroids on wound-healing reaction after glaucoma surgery. AB - PURPOSE: To determine and compare the effectiveness of octreotide, mitomycine-C and corticosteroids on wound-healing reaction after glaucoma surgery. METHODS: A full thickness scleral trephination was carried out by the same surgeon on tour groups of six rabbits. A sponge soaked in mytomicine-C was applied subconjunctivally in group 1 before trephination. Group 2 received corticosteroid drops tid topically for 14 days. Group 3 received subcutaneous octreotide injections tid for 14 days. The control group (group 4) was not given any drug that may interfere with wound healing. All groups received gentamycine drops tid for seven days. The rabbits were Sacrificed on the fourteenth day and the trephination area with overlying conjunctiva was excised. The samples were prefixed with glutaraldehyde, dehydrated and embedded in Araldite Cy 212. Ten semithin sections stained with toluidin blue were analysed for each group. Fibroblast and macrophage counts were performed on the surgical site and subconjunctival area. RESULTS: Intensive fibroblastic activity, increased number of vessels and active macrophages were observed only in group 4. The fibroblast and macrophage densities in this group were significantly higher than the other three groups in which wound healing was modulated (p < 0.001). Mean number of fibroblasts in group 1 was also significantly less than the ones of groups 2 and 3 (p < 0.01). Macrophage densities were similar in groups 1, 2 and 3. No statistical significance was found between groups 2 and 3 by means of fibroblast and macrophage densities. CONCLUSION: Octreotide reduced wound-healing reaction in a similar fashion to corticosteroids or mitomycine-C. These initial results seem promising. PMID- 14531625 TI - The natural history of juxtafoveal and subfoveal choroidal neovascularization in high myopia. AB - BACKGROUND: To assess the natural history of juxtafoveal and subfoveal choroidal neovascularization in eyes with high myopia. METHODS: We retrospectively reviewed the charts of 31 patients (31 eyes) with myopia > or = 6 diopters, well-defined juxtafoveal (1-200 microm from the foveal center) or subfoveal choroidal neovascularization (CNV) on fluorescein angiography at baseline, no prior laser treatment, age < or = 55 years and presenting visual acuity (VA) > or = 20/200. Initial and final VA were compared with the Wilcoxon signed rank test. Multifactor analysis of variance was used to assess the association between baseline characteristics of the lesion and final VA. RESULTS: Twenty-two patients were females and 9 males with a median age of 44 years (range 14-55). Median diopters spherical equivalent was -11.5 (range -6, -25). Follow-up ranged from 1 to 20 years (median, 3 years). Nine eyes had juxtafoveal CNV and 22 subfoveal involvement. Median final VA (20/100) was significantly worse than median initial VA (20/50)(p = 0.02). A decrease in VA > or = 2 lines occurred in 18 eyes, whereas 8 eyes remained stable and 5 improved (4 juxtafoveal membranes and 1 subfoveal membrane). Of the 9 juxtafoveal CNV, 7 had a final VA > or = 20/40 after a median follow-up of 4 years. By contrast, only 2 of the 22 subfoveal CNV had a final VA > or = 20/40 (median, 20/100) with a median follow-up of 2.5 years. The only factor associated with better final VA was the initial location of CNV (p = 0.0000). CONCLUSION: This study confirms the poor functional outcome of subfoveal CNV in degenerative myopia with more than 70% of patients having a final VA of 20/100 or less. Juxtafoveal CNV shows a better functional prognosis. These differences should be considered when planning treatment strategies. PMID- 14531626 TI - Fluorescein-assisted viscodissection for easier phacoemulsification. AB - TITLE: Fluorescein-assisted viscodissection for easier phacoemulsification. SETTINGS: Department of Ophthalmology, Cairo University. PURPOSE: The description of a new surgical technique to help overcome difficulties in the learning curve of phacoemulsification. METHODS: A mixture of Fluorescein 0.25% and methyl cellulose 0.25% is used for hydrodissection. Standard phacoemulsification is then continued until the initial groove reaches the plane between the lens cortex and the capsule when the fluorescein flows towards the anterior chamber in a Green Smoke. This sign indicates a proper depth for attempting to crack the nucleus into two halves. PMID- 14531627 TI - Ethmoidal sinus adenocarcinoma with orbital invasion. AB - PURPOSE: To report a rare case of massive ethmoidal adenocarcinoma with orbital invasion but minimal ophthalmic symptoms on presentation. METHODS: Case report of a 69-year-old, otherwise healthy, retired carpenter who was referred for treatment of bilateral senile cataract. RESULTS: A relative afferent pupillary defect and sectorial disc atrophy on ophthalmic examination led to further investigation and identification of an extensive ethmoidal neoplasm with orbital invasion. An incisional biopsy was performed and histopathologic examination revealed an adenocarcinoma of low-grade malignancy. CONCLUSION: Ethmoidal adenocarcinomas with orbital involvement may occasionally be relatively asymptomatic and masked by coexisting ocular disease. PMID- 14531628 TI - Variability of the normalised rim/disc area quotient estimated by laser scanning tomography. A comparison with conventional planimetry. AB - BACKGROUND: The introduction of the laser scanning tomograph raises the possibility of the estimation of optic disc morphology in a less subjective and less time-consuming way than the conventional planimetry of fundus photography. The aim of the present study was to estimate the efficiency of the HRT method in a comparison of data collected in a glaucomatous patient group to planimetric data obtained in young normal volunteers. METHODS: Repeated rim/disc area measurements were done in 10 glaucoma patients with a Heidelberg Retina Tomograph and in a group of five young volunteers with good vision. The intra-photographic, intra-observer examination of the planimetric method was compared to the inter photographic assessment of the HRT technique. RESULTS: The variability of the mean normalised area ratio quotient was nearly 50% less using laser scanning tomography techniques (mean rim/disc area ratio = 0.17). CONCLUSION: The results show the superiority of HRT over the conventional planimetric technique. Furthermore, the much shorter processing time and the advantages of a computer based databank in follow-up examination with HRT provides the hope that it could replace the rather time-consuming and subjective manual planimetric technique in the examination of glaucomatous optic nerve conditions. PMID- 14531629 TI - Ocular infections due to anaerobic bacteria. AB - The increased recovery of anaerobic bacteria in clinical infection has led to greater appreciation of these organisms in ocular infections. In studies that employed adequate method for recovery of anaerobes they were isolated from about a third of patients with conjunctivitis, half of the time in pure culture. The predominant recovered anaerobes were Clostridium spp., gram-negative anaerobic bacilli, and Peptostreptococcus spp. Anaerobic bacteria were also recovered from patients who wore contact lenses and developed conjunctivitis. Anaerobic bacteria were also reported in cases of keratitis. The most frequently recovered anaerobes were Propionibacterium spp., Peptostreptococcus spp., Clostridium spp., Prevotella spp., and Fusobacterium spp. The most frequently recovered anaerobes from dacryocystitis were Peptostreptococcus spp., Propionibacterium spp., Prevotella spp., and Fusobacterium spp. This review describes the microbiology, diagnosis and management of ocular infections due to anaerobic bacteria in children. PMID- 14531630 TI - Ophthalmic presentation of epidermoid cysts in an African community. AB - This retrospective study reviews the clinical and histopathological features of ocular epidermoid cyst. The 30-year laboratory records of all the patients of the Igbo ethnic group, whose surgical specimens were received at a central pathology laboratory service in south-eastern Nigeria, West Africa, were reviewed. The cases with the histopathologic diagnosis of epidermoid cyst involving ocular tissues were reviewed. The data analyzed included age at diagnosis, sex, location and size of cyst, clinical description of lesion and operative diagnosis. There were eighteen patients. Ten (55.6%) of these were males and 8 (44.4%) were females. Nearly three-quarters were below the age of 20 years. The cyst was present from birth in 4 (22.2%) patients and was acquired in the rest. It occurred on the external angle of the orbit in 7 patients; on the bulbar conjunctiva in 4 patients; in the lower fornix, upper lid and medial canthus in two patients each; and on the lower lid in one patient. There was no correct diagnosis of epidermoid cyst clinically, the most common misdiagnoses being dermoid cyst (55.6%) and conjunctival cyst (16.7%). Accordingly, epidermoid cyst should be included in the differential diagnosis of cystic lesions showing ophthalmic presentation. PMID- 14531631 TI - Combined phacoemulsification, intraocular lens implantation and trabeculectomy for chronic angle closure glaucoma. AB - PURPOSE: To investigate the results of combined phacoemulsification, intraocular lens implantation and trabeculectomy in Asian patients with chronic angle closure glaucoma (CACG). METHODS: This was a retrospective non-comparative case series of 55 consecutive patients (57 eyes) who underwent phacoemulsification, posterior chamber intraocular lens implantation and trabeculectomy for CACG at the Singapore National Eye Centre between 1997-1998. The surgical outcome was assessed in terms of intraocular pressure (IOP), the incidence of complications and the visual acuity at last follow-up. Success was defined as final IOP < or = 21 mmHg without medication and qualified success as final IOP < or = 21 mmHg with medication. Patients with final IOP > 21 mmHg who required further glaucoma surgery, lost light perception or became pthisical, were classified as failures. The eyes were further categorized into two groups according to whether single site or separate-site surgery was performed. The outcome was also compared among eyes in which per-operative antimetabolites were applied to the trabeculectomy site and those without antimetabolites. RESULTS: The mean follow up was 22.0 +/- 5.6 months (mean +/- SD). Success was achieved in 46 (81%) eyes, qualified success in 10 (17%) eyes, and failure in 1 (2%) eye. In terms of IOP outcome, the success rate was similar in the two surgical groups (single-site or separate site). There was no significant difference in IOP outcome among eyes in which per operative antimetabolites were used and eyes with no per-operative antimetabolites use. Forty-one eyes (72%) had 6/12 or better vision. There were no cases of intraoperative complications and the incidence of postoperative complications was low. CONCLUSIONS: Combined phacoemulsification, intraocular lens implantation and trabeculectomy is associated with good intraocular pressure control and visual outcome in patients with CACG. PMID- 14531632 TI - Visual outcome of the first 500 cataract extractions done in the field; Jimma, Ethiopia. AB - In 1998, a total of 617 cataract operations were performed on 523 patients at five outreach sites in Jimma Zone; south-western Ethiopia. The first 500 consecutive operations performed on 423 patients were analysed for visual outcome two weeks following field surgery. Four hundred and seventy-seven eyes underwent standard intra capsular cataract extraction (ICCE) while 23 eyes underwent extra capsular cataract extraction (ECCE). Seventy-seven patients underwent bilateral cataract surgery while 346 patients had been operated on only one eye. Forty-two patients (10.1%) or eighty-one eyes were lost for follow up. Thirty-nine of the 77 patients (50.6%) who received bilateral surgery and only 3 of 346 patients (0.9%) who received unilateral surgery were lost for follow up. Seventy-seven eyes (18.4%) had final visual acuity (v/a) of > or = 6/18; 388 eyes (92.6%) had v/a of > or = 6/60 and 395 eyes (94.3%) had v/a > or = 3/60. Capsule rupture and vitreous loss were the most common intra operative complications, accounting for 7.4% and 6%, respectively. Both complications were encountered in 1.8% of the eyes. Twenty-four eyes (5.7%) were blind despite surgery. However, only three eyes were blind as a result of complications related to surgery. There is a need for improvement in the screening of patients for surgery. The overall visual outcome obtained was satisfactory. Possible factors affecting visual outcome are considered and recommendations to effect improvement discussed. PMID- 14531633 TI - Eyelid fish-hook injury. AB - OBJECTIVE: To report a rare case of fish-hook eyelid injury. METHODS: Interventional case-report. A 25-year-old man presented with an upper eyelid fish hook wound. RESULTS: The hook was extracted under local anaesthesia using the back-out method. Postoperatively, no complications were observed. CONCLUSIONS: In the absence of serious ocular damage, the back-out method seems to be the best option for the removal of the hook in eyelid injuries. PMID- 14531634 TI - Selenium measurement in human plasma with Zeeman effect electrothermal atomic absorption spectrometry: sample stability and calibration method. AB - The dual aim of the present study is the investigation of the stability of plasma samples for selenium determination with time and temperature and the assessment of the calibration method. A comparative study is performed, using two calibration methods: standard addition to each sample and matrix matched curve. Our findings show that, in general, significant differences in the selenium content are observed when comparing the results obtained with these two calibration methods. Plasma samples stored at -20 degrees C are stable relative to the selenium content for a period of at least one year. PMID- 14531635 TI - Determination of zinc, copper, lead and cadmium in some medicinally important leaves by differential pulse anodic stripping analysis. AB - Levels of zinc, copper, lead and cadmium have been determined in some medicinally important leaves by differential pulse anodic stripping voltammetry (DPASV). High pressure digestion with nitric acid (HPA) was used for sample digestion. The accuracy of the method was verified by the parallel analysis of leaves with inductively coupled plasma atomic emission spectroscopy (ICP-AES) and recovery studies by the analysis of standard reference materials. Based on elemental levels the utility of these leaves in medicine are discussed. Statistical treatment has been used in order to understand the correlation between elements in these leaves. PMID- 14531636 TI - Element analysis and biological studies on ten oriental spices using XRF and Ames test. AB - Ten oriental spices were analyzed for their element composition using X-ray fluorescence (XRF): nutmeg (Myristica fragrans), coriander (Coriandrum sativum), safflower (Carthamus tinctorius), caraway (Carum carvi), Sicilian sumac (Rhus coriaria), aniseed (Anisum vulgare), black pepper (Piper nigrum), cardamom (Elettaria cardamomum), cumin (Cuminum cyminum) and nigella (Nigella sativum). The spices were found to contain the following elements: Mg, Al, Si, P, S, Cl, K, Ca, Ti, Mn, Fe, Cu and Zn, with varying concentrations. Mutagenic studies using Salmonella typhimurium strains TA97a, TA98, TA100, and TA102 showed that the above spices have no base pair substitution mutagenic activity. However, a weak frameshift mutagenicity has been shown by nutmeg and a very weak oxidative mutagenic action has been revealed by cumin. PMID- 14531637 TI - Development of zinc deficiency in 65Zn labeled, fully grown rats as a model for adult individuals. AB - The development of zinc deficiency in adults was studied in a metabolism experiment involving 31 adult, female rats labeled homogenously with 65Zn. The animals were fed restricted amounts (8 g/day) of a semisynthetic diet containing either 58 microgram Zn/g (control, n = 7) or 2 microgram Zn/g (Zn deficiency, n = 24). Control animals were sacrificed at day 0 (n = 3) and day 29 (n = 4). Zinc deficient animals were sacrificed at day 1, 2, 4, 7, 11, 16, 22, and 29 (3 animals per group). The development of zinc deficiency comprised 4 phases: (I) Fecal Zn excretion needed several days to adjust to the low level of Zn intake. The high initial Zn loss via feces was counterbalanced mainly by Zn mobilization from the skeleton. (II) During the 2nd week of deficiency Zn mobilization from tissue storage changed transiently to soft tissues (mainly muscle and fat tissue). (III) After the 2nd week the skeleton resumed to mobilize Zn. (IV) At the end of the study the skeleton Zn storage was exhausted and alkaline phosphatase activity indicated severe Zn deficiency. Urinary Zn excretion was too small to contribute quantitatively to changes in Zn metabolism during any phase of Zn deficiency. In conclusion, adults may compensate a deficient Zn supply by mobilizing tissue Zn for several weeks: The skeleton revealed to be the major short-term as well as long-term source of whole body tissue Zn that can be mobilized. PMID- 14531638 TI - Influence of zinc deficiency on the mRNA expression of zinc transporters in adult rats. AB - The accumulation of zinc in the cell is a sum of influx and efflux processes via transporter proteins, like the four Zn transporters (ZnT1-4), the divalent cation transporter 1 (DCT1) and of storage processes mainly bound to metallothionein (MT). To study the effect of Zn deficiency on mRNA expression levels, adult rats were used as an animal model. Food intake was restricted to 8 g/day containing 2 microg Zn/g fortified with pure phytate in Zn deficiency rats and 58 microg Zn/g in controls (n = 7). At day 1, 2, 4, 7, 11, 16, 22, and 29 of Zn deficiency, 3 animals were sacrificed, respectively (n = 24). Zn deficiency was evident from reduced plasma Zn, plasma alkaline phosphatase activity and severe mobilization of Zn from tissue stores (mainly skeleton), while food intake and body weight remained unaffected. Tissues representing Zn absorption (jejunum, colon), Zn storage and utilization (muscle, liver), and Zn excretion (kidney) were retrieved. Total RNA contents increased in colon (p = 0.003) and trend to decrease in liver (p = 0.086). Zn deficiency was without effect on tissue total RNA concentrations in muscle tissue and kidney. Real-time reverse transcription (RT) polymerase chain reaction (PCR) assays were developed and a relative quantification on the basis of GAPDH was applied. Assays allowed a relative and accurate quantification of mRNA molecules with a sufficiently high sensitivity and repeatability. All known Zn transporter subtypes were found in the tissues. ZnT3 was newly elucidated and sequenced in rat tissues. Expression patterns and reactions to Zn deficiency were specific for the tissue analysed. Expression results imply that some transporters are expressed constitutively, whereas others are highly regulated in tissues responsible for Zn homeostasis. The most distinct changes of expression levels were shown in colon which can therefore be postulated as a highly Zn sensitive tissue. MT was down-regulated in all tissues, massively in liver (p < 0.001) and in colon (p = 0.002) and in tendency also in the jejunum and kidney. In parallel with intracellular Zn status it is a potent candidate gene for Zn deficiency. ZnT1 and ZnT2 showed a significant up regulation of mRNA expression in colon (p = 0.032 and p = 0.026) and for ZnT2 a trend of down regulation in jejunum (p = 0.098). This study provides the first comparative view of regulation of gene expression and fully quantitative expression analysis of all known Zn transporters in a non growing adult rat model on a constant platform and therefore allows a direct comparison. PMID- 14531639 TI - Influence of the glass packing on the contamination of pharmaceutical products by aluminium. Part III: Interaction container-chemicals during the heating for sterilisation. AB - The interaction of chemicals with the container materials during heating for sterilisation was investigated, storing the components of parenteral nutrition solutions individually in sealed glass ampoules and in contact with a rubber stopper, and heating the system at 121 degrees C for 30 min. Subsequently, the aluminium content of the solutions was measured by atomic absorption spectrometry (AAS). The assay was also carried out with acids, alkalis and some complexing agents for Al. The containers were decomposed and also assayed for aluminium. 30 different commercial solutions for parenteral nutrition, stored either in glass or in plastic containers, were assayed measuring the aluminium present in the solutions and in the container materials. The results of all investigated container materials revealed an aluminium content of 1.57% Al in glass, 0.05% in plastic and 4.54% in rubber. The sterilisation procedure showed that even pure water was able to extract Al from glass and rubber, 22.5 +/- 13.3 microg/L and 79.4 +/- 22.7 microg/L respectively, while from plastic the aluminium leached was insignificant. The Al released from glass ampoules laid between 20 microg/L for leucine, ornithine and lysine solutions and 1500 microg/L for solutions of basic phosphates and bicarbonate; from rubber stoppers it reached levels over 500 microg/L for cysteine, aspartic acid, glutamic acid and cystine solutions. Ion exchange properties and influence of pH can explain the interaction of glass with some chemicals (salts, acids and alkalis), but only an affinity for aluminium could explain the action of some amino acids and other chemicals, as albumin and heparin, on glass and rubber, considering the aluminium release. Experiments with complexing agents for Al allowed to conclude that the higher the stability constant of the complex, the higher the Al release from the container material. PMID- 14531640 TI - Dietary intake of platinum and gold by children from Germany using duplicate portion sampling. AB - The dietary intake of platinum and gold by 84 small children, 42 boys and 42 girls at the age of 14 to 83 months, with different food consumption behaviour living in urban and rural areas of Germany was measured by the duplicate method with a seven day sampling period from May to September 1998. The levels in the food duplicates were in the range of < 0.01 to 450 ng Pt/kg (dry weight) (median: 22) and < 0.14 to 28 microg Au/kg (dry weight) (median: 0.645). Related to the body weight, Pt was in the range of < 0.81 to 32 ng/(kg (body weight) x week) (median: 2.3) and Au was < 0.015 to 2.6 microg/(kg (body weight) x week) (median: 0.068). Children consuming exclusively products from the super market showed slightly higher Pt concentrations in the food duplicates and a higher dietary intake per body weight than children with food consumption including products from the family owned vegetable gardens or the surrounding area and/or products from domestic animals of the surrounding area. No influence of the food consumption behaviour was found for the concentrations in the food duplicates or the dietary intake of Au. PMID- 14531641 TI - Dietary intake of arsenic, mercury and selenium by children from a German North Sea island using duplicate portion sampling. AB - The dietary intake of arsenic, selenium and mercury was studied for children living on the North Sea island Amrum, Germany. Altogether, 98 duplicate portions were collected from 14 children (age 1.5-5.5 years) in April and May 1995 over a sampling period of 7 days, respectively. The element concentrations in duplicate samples were measured by atomic absorption spectrometry. The weekly As and Hg intake (median and range) was 2.31 and 0.89-6.75 microg/(kg(bw) x week) for As and 0.13 and 0.060-0.62 microg/(kg(bw) x week) for Hg. Compared with the provisional tolerable weekly intake (PTWI) for As and Hg as proposed by the WHO, German children from coastal areas reveal no health risks due to As and Hg dietary intake. The daily Se intake (median and range) was 19 and 6-160 microg/day. The appropriate Se intake of 10-40 microg/day, as recommended by the Austrian, German, and Swiss Nutrition Councils for 1-4 years-old children, was not reached in 8 out of 49 cases (16.3%), whereas the recommended Se intake, fixed at 15-45 microg/day for the 4-7 years-old children, was not reached in 15 out of 48 cases (31.3%). PMID- 14531643 TI - Effect of lactoferrin addition on the dialysability of iron from infant formulas. AB - A possible enhancing effect of lactoferrin (Lf) on iron absorption by breast-fed infants has been suggested, however the available results failed to confirm this hypothesis. Nevertheless, Lf could be useful in protecting the lipid fraction of infant formulas against oxidation. Concerning the possibility of adding Lf to infant formulas with this aim, we considered it necessary to evaluate the effect of this addition on iron dialysability, which was used as a parameter indicator of bioavailability. An in vitro dialysability method was applied to three types of infant formulas, with and without Lf added, respectively. In none of the analysed formulas did the added Lf have a negative effect on iron dialysability, and in only two of them (adapted formulas) was a statistically significant (p < 0.05) increase observed, although of low practical significance value. In conclusion, iron dialysability, used as an estimate of bioavailability, seems to be neither enhanced nor lowered by Lf addition to infant formulas. PMID- 14531642 TI - Selenium modifies glutathione peroxidase activity and glutathione concentration in mice exposed to ozone-provoked oxidative stress. AB - The aim of this study was to show the direct effect of selenium on glutathione peroxidase (GSH-Px) activity and GSH/GSSG concentrations in 3- and 6-month-old mice. An ozone-oxygen mixture was used to provoke an oxygen stress. To measure the Se-effect mice were gavaged with sodium selenite. GSH-Px activity and total glutathione concentrations were determined in serum and in the postnuclear fraction of liver and lungs. Additionally glutathione concentrations were determined in whole blood. Both ozone and selenium, administered separately, reduced GSH-Px activity in lungs of 6-month-old animals, while in young mice an opposite effect of Se was observed. Ozone administered jointly with Se did not influence GSH-Px activity in 6-month-old mice, while in young, 3-month-old mice, a stimulatory effect in lungs was observed. There were no significant changes in GSH-Px activity in the liver of 6-month-old mice, but the stimulatory effect occurred in young mice treated with Se and Se & ozone jointly. In young mice, ozone (also ozone with Se) augmented glutathione concentrations. The response to ozone and selenium strictly depended on age and the antagonism between selenium and ozone was observed only in a few cases. PMID- 14531644 TI - Large tissue freezing. PMID- 14531645 TI - Cycle characteristics of day 3 embryo transfers with 4-cell embryos only. AB - PURPOSE: Patient and cycle characteristics of day 3 transfers with developmentally lagging 4-cell embryos only were analyzed and related to the outcome of a live birth. METHODS: Day 3 transfers with either 4-cell embryos only (study group; n = 138) or 8-cell embryos only (control group; n = 282) were compared retrospectively. RESULTS: The total dose of FSH per treatment was higher, while the number of oocytes, zygotes, and transferred embryos was lower in the study group cycles compared to controls. The implantation, pregnancy, and live birth rates were dramatically lower in the study group compared to the control group. In the study group, the few cycles resulting in a live birth were characterized by a normal ovarian response to stimulation, similar to that of control group cycles with- or without a live birth. CONCLUSIONS: In cycles characterized by intensive ovarian stimulation, but poor response, the chance for a live birth is extremely low after day 3 transfer of 4-cell embryos. PMID- 14531646 TI - The effect of preincubation period of oocytes on nuclear maturity, fertilization rate, embryo quality, and pregnancy outcome in IVF and ICSI. AB - PURPOSE: To clarify the effect of preincubation of oocytes on the results of IVF and ICSI. METHODS: A total of 176 IVF and 64 ICSI cycles received long protocol ovarian stimulation. The oocytes were incubated for 1-8 h before insemination or sperm injection. Metaphase II (MII) percentage was evaluated in the ICSI arm; fertilization rates, embryo quality, and pregnancy outcomes were analyzed in both IVF and ICSI arms according to the preincubation period duration of oocytes. RESULTS: The MII percentage of the ICSI arm was significantly lower (P < 0.05) in the group with preincubation period of < 2.5 h. The fertilization rates in groups with preincubation for 2.5-5.5 h were significantly higher (P < 0.001) for IVF. Embryo quality and pregnancy outcomes were not significantly different between the IVF or ICSI arm. CONCLUSIONS: The preincubation of oocytes for at least 2.5 h is beneficial to both IVF and ICSI outcomes by increasing the nuclear maturity of oocytes. PMID- 14531647 TI - Does surgical management of endometriosis within 6 months of an in vitro fertilization-embryo transfer cycle improve outcome? AB - PURPOSE: To assess the effect on cycle outcome of surgery for endometriosis performed in the 6 months prior to in vitro fertilization-embryo transfer (IVF ET). METHODS: Retrospective analysis of all patients with a primary diagnosis of endometriosis undergoing IVF-ET during 12 consecutive months. The setting included tertiary care assisted reproductive technology program. Ninety-five consecutive cycles in candidates for autologous IVF-ET who had undergone surgical resection of endometriosis within 60 months of oocyte retrieval were evaluated. Five patients with persistent or recurrent endometriomas > 3 cm at the time of IVF-ET were excluded. Patients were divided into two groups based on the interval between the most recent surgical intervention and oocyte aspiration-Group I: < or = 6 months' interval (19 cycles); Group II: > 6 months' interval (76 cycles). RESULTS: Mean intervals between surgery and oocyte aspiration were significantly different between the groups (Gr I: 3.7 +/- 0.3 months; Gr II: 21.8 +/- 1.6 months; p < 0.001). There were no significant differences between groups with regards to age, extent of endometriosis, or results of ovarian stimulation and oocyte aspiration. Ongoing pregnancy rates (Gr I: 63.2%; Gr. II: 60.5%) were similar. Regression analyses revealed no correlation between implantation rates and either endometriosis score (r = -0.09) or surgery-oocyte aspiration interval (r = -0.13). CONCLUSIONS: The interval between surgical management of endometriosis and oocyte aspiration did not have an impact on IVF-ET cycle outcome in the absence of significant persistent or recurrent ovarian disease. Any effect of surgery on enhancing spontaneous conception may be overcome by the inherently greater impact of IVF-ET on implantation and pregnancy. PMID- 14531648 TI - Estimation of second polar body retention rate after conventional insemination and intracytoplasmic sperm injection: in vitro observations from more than 5000 human oocytes. AB - PURPOSE: Tripronucleate (3pn) development after conventional insemination (CONV) or ICSI was analyzed to estimate the rate of second polar body retention giving rise to 3pn formation. METHODS: Data from 453 consecutive IVF cycles were reviewed during a 6-month period. Mature oocytes were monitored in ICSI (n = 3195) and CONV (n = 2274) groups by fertilization assessment 16-18 h post insemination. Ovulation induction protocols and in vitro culture conditions remained constant during the study interval. RESULTS: Normal (2pn) fertilization occurred in 74.2% and 70.5% for CONV and ICSI groups, respectively (p < 0.003). 1pn formation was observed in 4.5% of CONV oocytes, and 2.5% of ICSI oocytes (p < 0.001); 3pn formation was 8.1% in the CONV group, and 2.5% in the ICSI group (p < 0.0001). We observed 4pn formation in 0.4% of oocytes in the CONV group, but in only 0.04% of oocytes fertilized with ICSI (p < 0.007). Cellular degeneration occurred in 2.4% of oocytes inseminated conventionally, and in 3.5% of oocytes fertilized by ICSI (p = 0.02). Maternal age did not impact pronuclear status. CONCLUSIONS: We found the 3pn formation rate after ICSI to be approximately one third that observed in the CONV group. Extrapolating the ICSI data to the CONV data, it may be inferred that 2.5% of 3pn development after CONV was due to second polar body retention. This suggests that 5.6% of CONV oocytes showed dispermic fertilization. Decreasing oocyte quality with increasing maternal age had no apparent influence on any of the fertilization outcomes. PMID- 14531649 TI - Intracytoplasmic sperm injection and conventional in vitro fertilization are complementary techniques in management of unexplained infertility. AB - PURPOSE: To evaluate the role of ICSI in unexplained infertility. METHODS: In 125 cycles with six or more oocytes retrieved per cycle, sibling oocytes were randomly allocated to IVF or ICSI (group A). In 74 cycles with less than six oocytes retrieved per cycle, cycles were allocated to IVF or ICSI (group B). RESULTS: In group A, ICSI fertilization rate of 61% per allocated oocyte was higher than IVF fertilization rate of 51.6% (P < 0.001). Complete fertilization failure occurred in 19.2 and 0.8% of cycles in IVF and ICSI, respectively (P < 0.001). In group B, fertilization rate in IVF cycles was 53.3% as compared to 60.7% per allocated oocyte in the ICSI cycles (P = 0.29). Complete fertilization failure was higher (P = 0.02) in conventional IVF (34.3%) than ICSI cycles (10.3%). CONCLUSIONS: Allocation of sibling oocytes to IVF and ICSI in the first cycle minimizes risk of fertilization failure. For patients with limited number of oocytes, ICSI technique is recommended. PMID- 14531650 TI - Birth of healthy twins resulting from donated oocytes and posthumous use of frozen-thawed spermatozoa obtained prior to chemotherapy. AB - The improvement in life expectancy, following better cancer therapy combined with new options in treating male infertility have increased the use of sperm freezing. We describe a rare case of twin pregnancy of a childless widow, using donated oocytes after ICSI with her deceased husband's banked frozen spermatozoa. Sperm was frozen before chemotherapy treatment and a written consent for future thawing and injection to donor oocytes was given. Use of thawed spermatozoa after death was not mentioned in the consent form. During the husband's illness years, ICSI of donated oocytes fertilized with thawed spermatozoa yielded three embryos, but no pregnancy was achieved after ET. While waiting for a subsequent oocyte donation, the male partner died of his malignancy. The woman expressed her wish to be treated with the frozen spermatozoa of her deceased husband. Since the original informed consent did not cover postmortem use of the husband's sperm, a special application for its use was made and subsequently approved by the Israeli Ministry of Health legal advisor. Another six donated oocytes were fertilized with the deceased's thawed spermatozoa, and the transfer of four embryos to the uterine cavity resulted in a twin pregnancy and later delivery of two healthy babies. This case emphasizes the importance of written informed consent signed by patients with a life-threatening disease at the time of banking spermatozoa, and discussing the possibility of its posthumous use. Several legal and ethical concerns are discussed. PMID- 14531652 TI - Recurrent empty follicle syndrome. AB - We report a case of a recurrent empty follicle syndrome. The patient was admitted to our intracytoplasmic injection program because of her partner's azoospermia. Ovarian stimulation was accomplished using gonadotrophin therapy after treatment with oral contraceptive pills followed by gonadotrophin-releasing hormone agonist. Thirty-six hours after the administration of HCG (human chorionic gonadotrophins), transvaginal oocyte retrieval yielded no oocytes despite the aspiration and flushing of all available follicles. Two years later, a second treatment cycle was started using the same pituitary desensitisation and ovarian stimulation regimens. HCG from a different batch with respect to that used in the first treatment cycle was administered. Aspiration and repeated flushing of all follicles of one ovary failed to yield any identifiable oocyte. The beta-HCG and progesterone serum concentrations on the day of retrieval were 181 mIU/mL and 3.79 ng/mL, respectively. New oocyte retrieval was planned 6 h after the first attempt for aspiration of follicles. Again, no ova were obtained at this second trial despite the aspiration of the all follicles. As to our knowledge this is the first report of recurrent EFS (empty follicle syndrome) and managed without repeating the HCG injection on the day of unsuccessful oocyte retrieval. PMID- 14531653 TI - Noncyclic [10-S-5] sulfuranide dioxide salts with three S-C bonds: a new class of stable hypervalent compounds. AB - Phenyl triflate reacts with CF3SiMe3/Q+F- (Q+ = [K(18-crown-6)]+, Me4N+) and (Me2N)3S+Me3SiF2- to afford the first noncyclic [10-S-5] sulfuranide dioxide salts, [(CF3)3SO2]-Q+, with three S-C bonds, whose molecular structure was determined by X-ray crystallography. PMID- 14531651 TI - Molecular and cytogenetic characterization of two azoospermic patients with X autosome translocation. AB - PURPOSE: To report two azoospermic patients with reciprocal X-autosome translocations. METHODS: Cytogenetic analysis utilizing GTG-banding and Yq microdeletions shown by polymerase chain reaction (PCR) with 12 sequence-tagged site (STS) markers for Y chromosome microdeletions. RESULTS: Cytogenetic analysis showed one man with 46,Y,t(X;19)(q22;q13.3) and the other with 46,Y,t(X;8)(p22;q11). Neither had any Yq microdeletions shown. The patient with 46,Y, t(X;8)(p22;q11) showed a slightly lower than normal testosterone level. By NCBI-Blast search, we found four testis-specific genes, t-complex-associated testis-expressed 1-like (TCTE1L), Ferritin, heavy polypeptide-like 17 (FTHL17), Testis expressed sequence 13A (TEX13A), and Testis expressed sequence 13B (TEX13B) located near breakpoints on X chromosome. FTHL17, TEX13A, and TEX13B are spermatogonially-expressed, germ-cell-specific genes. CONCLUSION: This is the first clinical report of azoospermia with reciprocal X-autosome translocations on Xp22 and q22. These translocations on Xp22 and q22 may be direct genetic risk factors for azoospermia. PMID- 14531654 TI - Inter- and intramolecular activation of aromatic C-H bonds by diphosphine and hydrido-bridged dinuclear iridium complexes. AB - Reactions of [(Cp*Ir)2(mu-dmpm)(mu-H)2]2+ (1) with NaOtBu in aromatic solvent at room temperature give [(Cp*Ir)(H)(mu-dmpm)(mu-H)(Cp*Ir)(Ar)]+ [Ar = Ph (3), p-Tol (4a), m-Tol (4b), 2-furanyl (5a), 3-furanyl (5b)] via intermolecular aromatic C-H activation. Treatment of [(Cp*Ir)2(mu-dppm)(mu-H)2]2+ (2) with base (Et2NH) results in intramolecular C-H activation of the phenyl group in the dppm ligand to give [(Cp*Ir)(H){mu-PPh(C6H4)CH2PPh2}(mu-H)(Cp*Ir)]+ (6). The structures of 3, 5a, and 6 have been determined by X-ray diffraction methods. PMID- 14531655 TI - Fluorogenic resolution of ligand binding by a nucleic acid aptamer. AB - An ATP-binding nucleic acid aptamer was converted into a solution-phase fluorescence-detected sensor by incorporating a 2'-amine group in a structure that is sensitive to a ligand-induced conformational change in the aptamer. The 2'-amine substitution in the free aptamer reacts efficiently with fluorescamine to yield a fluorescent product detectable by energy transfer to a tethered fluor emitting at 615 nm. In the presence of ATP, the fluorogenic reaction is substantially reduced, both under simplified conditions and in a urine background. Fluorogenic chemistry represents a potentially general approach for creating sensors from aptamers that bind their ligands via induced fit. PMID- 14531656 TI - Highly sensitive in vitro selections for DNA-linked synthetic small molecules with protein binding affinity and specificity. AB - We have developed in vitro selections for DNA-linked synthetic small molecules with protein binding affinity and specificity. These selections require only generally accessible equipment, offer high degrees of enrichment of active molecules from mixtures of predominantly inactive species, can be applied to a variety of unrelated proteins, and require approximately 108-fold less material than existing synthetic molecule screening methods. Iterating these selections multiplies the net enrichment of active molecules, enabling enormous overall enrichment factors exceeding 106 to be achieved. Further, the selections can be adapted to select for binding specificity in addition to binding affinity. The application of methods described in this work may play a key role in the discovery of desired molecules from DNA-templated synthetic libraries. PMID- 14531657 TI - Cellular uptake of aminoglycosides, guanidinoglycosides, and poly-arginine. AB - Aminoglycosides (including neomycin B and tobramycin) exhibit poor uptake by eukaryotic cell lines. When the amines of these natural products are converted into guanidine groups, their cellular uptake is dramatically enhanced. We have synthesized BODIPY-containing aminoglycosides and guanidinoglycosides to evaluate their cellular uptake properties. Fluorescence activated cell sorting (FACS) and fluorescence microscopy are used to compare the membrane translocation and the cellular localization of these compounds. Upon guanidinylation, the cellular uptake efficiencies of tobramycin and neomycin B are enhanced by 10-fold and 20 fold, respectively. Guanidino-neomycin B exhibits a highly efficient uptake, superior to a fluorescent poly-arginine peptide. Interestingly, the cellular uptake of this common transduction peptide is inhibited by guanidine-neomycin B, suggesting a similar uptake mechanism for both the arginine-rich peptides and the guanidinoglycosides. PMID- 14531658 TI - A new fluoride selective fluorescent as well as chromogenic chemosensor containing a naphthalene urea derivative. AB - A new naphthalene derivative containing a urea group at the 1,8-position of naphthalene was synthesized and showed a unique absorption and fluorescence peak with a fluoride ion. Calculations suggested that a new peak was attributed to the increased anion character of urea nitrogen due to the strong interaction of the fluoride and N-H protons. PMID- 14531659 TI - Understanding the mechanism of H(+)-induced demetalation as a design strategy for robust iron(III) peroxide-activating catalysts. AB - The FeIII-TAML (tetra-amido macrocyclic ligand) activators 1 (Y = Cl) and 2 (Y = H2O), a (R = Me, X = H), b (Me, Cl), c (Me, MeO), d (Et, Cl), e (F, H), f (F, Cl), are five-coordinated in the solid state (X-ray crystallography) but are six coordinated species in water with two H2O axial ligands. The first pKa's of aqueous ligands are in the range of 9.5-10.5. The acid-induced demetalation of 2 follows the equation kobs = k1*[H+] + k3*[H+]3. The rate constants k1* and k3* vary by 5 and 11 orders of magnitude depending on the nature of substituents R. The highest stabilization against the demetalation is achieved for R = F. PMID- 14531660 TI - Stereospecific radical polymerization of N-triphenylmethylmethacrylamides leading to highly isotactic helical polymers. AB - N-Triphenylmethylmethacrylamide (TrMAM) and N-[di(4 butylphenyl)phenylmethyl]methacrylamide (DBuTrMAM) were polymerized using radical initiators. The polymer PTrMAM obtained from TrMAM was insoluble in the common solvents, but was dissolved in sulfuric acid to quantitatively give polymethacrylamide. The 1H NMR spectrum of PTrMAM in sulfuric acid-d2 indicated that the derived polymethacrylamide was nearly 100% isotactic. On the other hand, DBuTrMAM yielded the polymer PDBuTrMAM, which was soluble in THF and chloroform and whose NMR pattern in sulfuric acid-d2 was similar to that of PTrMAM. The polymerization of TrMAM and DBuTrMAM in an optically active solvent, (-)- or (+) menthol, afforded the optically active polymers having a prevailing one-handed helical structure. The monomers were not solvolized in a methanol-chloroform mixture. TrMAM was not polymerized by anionic initiators. PMID- 14531661 TI - Structure of antibacterial peptide microcin J25: a 21-residue lariat protoknot. AB - The antibacterial peptide microcin J25 (MccJ25) inhibits bacterial transcription by binding within, and obstructing, the nucleotide-uptake channel of bacterial RNA polymerase. Published covalent and three-dimensional structures indicate that MccJ25 is a 21-residue cycle. Here, we show that the published covalent and three dimensional structures are incorrect, and that MccJ25 in fact is a 21-residue "lariat protoknot", consisting of an 8-residue cyclic segment followed by a 13 residue linear segment that loops back and threads through the cyclic segment. MccJ25 is the first example of a lariat protoknot involving a backbone-side chain amide linkage. PMID- 14531662 TI - Large oriented arrays and continuous films of TiO(2)-based nanotubes. AB - We report for the first time a one-step, templateless method to directly prepare large arrays of oriented TiO2-based nanotubes and continuous films. These titania nanostructures can also be easily prepared as conformal coatings on a substrate. The nanostructured films were formed on a Ti substrate seeded with TiO2 nanoparticles. SEM and TEM results suggested that a folding mechanism of sheetlike structures was involved in the formation of the nanotubes. The oriented arrays of TiO2 nanotubes, continuous films, and coatings are expected to have potentials for applications in catalysis, filtration, sensing, photovoltaic cells, and high surface area electrodes. PMID- 14531663 TI - 2,3,4- or 2,3,5-trisubstituted furans: catalyst-controlled highly regioselective ring-opening cycloisomerization reaction of cyclopropenyl ketones. AB - 2,3,4- or 2,3,5-trisubstituted furans were highly regioselectively formed from the cycloisomerization reaction of the same starting cyclopropenes 1 via the subtle choice of the transition metal halides. Under the catalysis of 5 mol % PdCl2(CH3CN)2, 2,3,5-trisubstituted furans 2 were given in 50-88% yields with 95 99% regioselectivities, while 2,3,4-trisubstituted furans 3 were formed in 78-96% yields with 99% regioselectivities under the catalysis of 5 mol % CuI. PMID- 14531664 TI - Heteroepitaxial growth of a zeolite film with a patterned surface-texture. AB - We focus on the eteroepitaxial growth of trigonal chabazite with voids of three dimensional intersection structures. The differences in the structures originate from the stacking sequences of the six-membered rings. Sodalite along 111 is constructed by an "abcabc" stacking sequence, while chabazite along [111] is constructed by "abab". Therefore, chabazite can grow heteroepitaxially on a sodalite substrate while keeping the relation of chabazite (111)//sodalite {111}. In this Communication, we report on the first success of the heteroepitaxial growth of a continuous zeolite (chabazite) film with a unique patterned surface texture on a millimeter-sized sodalite single crystal. We have found that the unique texture of chabazite films is rationalized by the heteroepitaxial growth of chabazite on sodalite and the subsequent twin formation of chabazite. PMID- 14531665 TI - Targeting the transcriptional machinery with unique artificial transcriptional activators. AB - The link between a growing number of human diseases and misregulation of gene expression has spurred intense interest in artificial transcriptional activators that could be used to restore controlled expression of affected genes. To expand the repertoire of activation domains available for the construction of artificial transcriptional regulators, a selection strategy was used to identify two unique activation domain motifs. These activation domains bear little sequence homology to endogenous counterparts and bind to unique sites within the transcriptional machinery. A comparison with two well-characterized activation domains, VP2 and P201, demonstrated for the first time that functional potency is not solely dictated by binding affinity. Finally, the selection strategy described is readily applicable to the identification of small molecule activation domains. PMID- 14531666 TI - Synthesis of monodisperse polymers from proteins. AB - Proteins are functional biopolymers; viewed as molecules, they are also monodisperse polyamides with chemically reactive side chains. This paper describes the use of proteins as starting materials for the synthesis of monodisperse polymers with nonbiological functionalities attached to the side chains. It demonstrates the complete derivatization of amine groups (lysine side chains and N-termini) on three different proteins by addition of activated carboxylate reagents in aqueous solutions containing sodium dedecyl sulfate (SDS), under denaturing conditions. Several different acylating reagents were used to generate derivatized proteins; the resulting compounds constitute a new class of monodisperse, semisynthetic polymers, having the potential for wide variation in the structure of the backbone and of the side chains. Modification of lysozyme on a gram scale demonstrated that the method can generate useful quantities of material. PMID- 14531667 TI - Amine-catalyzed coupling of allenic esters to alpha,beta-unsaturated carbonyls. AB - Nucleophilic amines catalyze the 1,4-addition of allenic esters to alpha,beta unsaturated carbonyl compounds. The reactions proceed under mild conditions. In addition, Baylis-Hillman products serve as appropriate electrophiles for the process such that a one-pot, three-component coupling of aldehydes, acrylates, and allenic esters may be achieved. PMID- 14531668 TI - Photoinduced magnetization of the cyano-bridged 3d-4f heterobimetallic assembly Nd(DMF)(4)(H(2)O)(3)(micro-CN)Fe(CN)(5).H(2)O (DMF = N,N-dimethylformamide). AB - Photoinduced magnetization of the cyano-bridged 3d-4f heterobimetallic assembly Nd(DMF)4(H2O)3(mu-CN)Fe(CN)5.H2O (1) (DMF = N,N-dimethylformamide) is described in this paper. The chiMT values are enhanced by about 45% after UV light illumination in the temperature range of 5-50 K. We propose that UV light illumination induces a structural distortion in 1. This small structural change is propagated by molecular interactions in the inorganic network. Furthermore, the cooperativity resulting from the molecular interaction functions to increase the activation energy of the relaxation processes, which makes observation of the photoexcited state possible. The flexible network structure through the hydrogen bonds in 1 plays an essential role for the photoinduced phenomenon. This finding may open up a new domain for developing the molecule-based magnetic materials. PMID- 14531669 TI - Reaching into the major groove of B-DNA: synthesis and nucleic acid binding of a neomycin-hoechst 33258 conjugate. AB - Synthesis of a neomycin-Hoechst 33258 conjugate is reported. The conjugate combines the ligands known to selectively bind in the duplex and the triplex groove. The conjugate stabilizes DNA duplex over DNA triplex. The conjugate selectively stabilizes the DNA duplex (in the presence of salt), with as little as 2 muM of the ligand leading to a DeltaTm of 25 degrees C. PMID- 14531670 TI - Single-crystal-to-single-crystal topochemical polymerizations of a terminal diacetylene: two remarkable transformations give the same conjugated polymer. AB - Host-guest chemistry was used to prepare a cocrystal of a dipyridyl substituted oxalamide host and a resorcinol derivative of a terminal diacetylene. X-ray crystallography revealed that the molecules assemble into a triple-helix with the diacetylene functionalities aligned in the center of the helix. Upon heating, the diacetylenes polymerize to give the corresponding polymer. In a second experiment, the X-ray structure of the crystalline hydrate of the same diacetylene also showed a suitable alignment of the diacetylene functionalities. These crystals polymerize at only 50 degrees C with the C1 carbon end of the diacetylene undergoing an unprecedented 2.4 A inward swinging movement. This results in a remarkable 9% increase in crystal density. These are the first structurally characterized polymerizations of any terminal diacetylene. A detailed comparison of the two sets of structural changes offers an interesting insight into the precise trajectories of polymerization reactions. PMID- 14531671 TI - Improving the performance of methylalumoxane: a facile and efficient method to trap "free" trimethylaluminum. AB - The presence of "free" trimethylaluminum (TMA) in methylalumoxane (MAO) solutions can be highly detrimental to the performance of metallocene and "post metallocene" olefin polymerization catalysts. The most used strategy to remove "free" TMA is to evaporate MAO solutions to dryness, until a free-flowing white powder ("solid MAO") is left. This procedure is tedious and potentially hazardous, because in some cases the distillate is a concentrated hydrocarbon solution of TMA. Moreover, "solid MAO" is poorly soluble in common polymerization media, and once in solution it can regenerate TMA to some extent. This communication reports on a facile alternative, which consists in the controlled addition of a sterically hindered phenol, such as 2,6-di-tert-butylphenol, effectively trapping "free" TMA. We show here that 2,6-di-tert-butylphenol/MAO solutions activate equally well the dichloro-precursors of well-known zirconocene and bis(phenoxyimine)Ti catalysts, and that their use in propene polymerization results in a substantially higher productivity, polymer stereoregularity, and/or average molecular mass compared with activation by MAO alone. PMID- 14531672 TI - Gas-phase peptide/protein cationizing agent switching via ion/ion reactions. AB - Polypeptide ions comprising different cationizing agents show distinct fragmentation behavior in the gas phase. Thus, it is desirable to be able to form ions with different cationizing agents such as protons and metal ions. Usually, metal-cationized peptide/protein ions are introduced to the mass spectrometer by electrospraying solutions containing a mixture of the peptide/protein of interest and a metal salt. A new technique for generating metal-containing polypeptide ions that involves gas-phase ion/ion reactions is described. In this strategy, solutions of metal-containing ions and solutions of proteins are each electrosprayed into separate ion sources. The approach allows for independent maximization of ion signal and selection of ions prior to gas-phase reactions. Selected ions are stored in a quadrupole ion trap where reactions of ions of opposite polarity form metal-cationized peptides and proteins in the gas phase by cation switching. This approach affords a high degree of flexibility in forming metal-containing peptide and protein ions via the ability to mass-select reactant ions. The ability to form a variety of peptide/protein ions with various cationizing reagents in the gas phase is attractive both for the study of intrinsic interactions of metal ions with polypeptides and for maximizing the structural information available from tandem mass spectrometry of peptides and proteins. PMID- 14531673 TI - A palladium-catalyzed glycosylation reaction: the de novo synthesis of natural and unnatural glycosides. AB - A highly stereoselective and sterospecific palladium-catalyzed glycosylation reaction of a variety of alcohols is reported. The reaction selectively converts alpha-2-substituted 6-carboxy-2H-pyran-3(6H)-ones into alpha-2-substituted 6 alkoxy-2H-pyran-3(6H)-ones with complete retention of configuration and similarly converts the pyranones with beta-carboxy groups into pyranones with beta-alkoxy groups. The reaction works equally well with both amino acid- and carbohydrate based alcohols. To demonstrate the utility of this process for carbohydrate chemistry several of the products were selectively converted into alpha-manno pyranosides in two additional steps. Because the 2-substituted 6-carboxy-2H-pyran 3(6H)-ones are prepared by asymmetric synthesis, this reaction can be used for the preparation of either d- or l-pyranones. PMID- 14531674 TI - Structure of colloidal bismuth subcitrate (CBS) in dilute HCl: unique assembly of bismuth citrate dinuclear units ([Bi(cit)(2)Bi](2-)). AB - Colloidal bismuth subcitrate (CBS, De-Nol) has been used for several decades for the treatment of gastric and duodenal ulcers, and Helicobacter pylori infection together with antibiotics. The solubility of CBS in water is found to be dramatically affected by pH, from >70 mg/mL at pH 7 to only about 1 mg/mL at pH 3. CBS was crystallized in dilute HCl at pH 3, and unique assembly of three basic bismuth citrate dimeric units ([Bi(cit)2Bi]2-) leads to the formation of two dimensional sheets and 3D polymer. PMID- 14531675 TI - DNA-templated metal catalysis. AB - Hydrolysis of an ester substrate by a CuII complex catalyst, both attached to oligo-peptide nucleic acids (PNA), is accelerated up to 485-fold in the presence of a complementary DNA template. The approach combines the sequence selectivity of DNA-templated reactions with signal amplification by multiple turnover and the versatility of metal catalysis. PMID- 14531676 TI - Catalytic asymmetric rearrangement of allylic trichloroacetimidates. A practical method for preparing allylic amines and congeners of high enantiomeric purity. AB - COP-Cl catalyzes the rearrangement of (E)-allylic trichloroacetimidates to provide transposed allylic trichloroacetamides of high enantiopurity, a transformation that underlies the first truly practical method for transforming prochiral allylic alcohols to enantioenriched allylic amines and congeners. The high functional group compatibility of this asymmetric rearrangement and the demonstrated broad utility in synthesis of the allylic trichloroacetimidate to allylic trichloroacetamide conversion are singular features of this new catalytic asymmetric reaction. PMID- 14531677 TI - Active-site coating for molecular discrimination in heterogeneous catalysis. AB - A coherent layer of zeolite A has been applied to a SiO2-supported Pt-Fe oxidation catalyst, using total surface charge-reversal. The zeolite has tracked the metals into the pore structure of the support, covering the active sites. The zeolite channel size is large enough to allow access of CO and O2 to the active sites, and to allow CO2 to emerge, but it excludes larger molecules. The presence of the zeolite membrane transforms the supported Pt-Fe into a highly specific catalyst, which can discriminate between CO and butane, even after the macroscopic catalyst particles are crushed. PMID- 14531678 TI - Self-promoted electron transfer from cobalt(II) porphyrin to p-fluoranil to produce a dimer radical anion-cobalt(III) porphyrin complex. AB - Self-promoted electron transfer from a cobalt(II) porphyrin [Co(II)OEP] to p fluoranil (F4Q) occurs, exhibiting a second-order dependence of the electron transfer rate with respect to the F4Q concentration due to the formation of a strong complex between the dimer radical anion [(F4Q)2*-] and the resulting Co(III)OEP+. PMID- 14531679 TI - Laser flash photolysis formation and direct kinetic studies of manganese(V)-oxo porphyrin intermediates. AB - Irradiation of porphyrin-manganese(III) perchlorate complexes in acetonitrile with 355 nm laser light gave MnV-oxo intermediates that were characterized by their UV-vis spectra and reactivities. The MnV-oxo species of tetrakis(pentafluorophenyl)porphyrin (2), tetraphenylporphyrin (4), and tetra-(4 (N-methylpyridiniumyl))porphyrin (6) were generated. Second-order rate constants for reactions of 2 with substrates were as follows: 6.1 x 105 M-1 s-1 (cis stilbene), 1.3 x 105 M-1 s-1 (diphenylmethane), 1.3 x 105 M-1 s-1 (ethylbenzene), and 0.55 x 105 M-1 s-1 (ethylbenzene-d10). In oxidations of cis-stilbene and diphenylmethane, the order of reactivity of the MnV-oxo species was 2 > 6 > 4. PMID- 14531680 TI - Synthesis of poly(biphenylene ruthenacyclopentatrienylene), a new organometallic conducting polymer with ferromagnetic interaction in its reduced state. AB - A ruthenacyclopentatriene polymer 2 was synthesized by metallacycling polymerization (MCP). Polymer 2 with Mn = 3400 and Mw = 5800 consisted of identical ruthenacyclopentatriene units. The UV-vis spectra of 2 showed a shift of the pi-pi* band to a longer wavelength due to the enlargement of the pi conjugation. Polymer 2 underwent reversible reduction due to the ruthenacycle moiety at -1.01 V versus ferrocenium/ferrocene. The EPR spectrum of the reduced form of 2 in frozen THF indicated ferromagnetic interaction of spins at ruthenacyclopentatriene units. PMID- 14531681 TI - Methyl/ethyl differentiation in diastereoselective hydride addition to [Tp'W(CO)(PhCCMe)(NH=CMeEt)][BAr'(4)]. AB - Hydride addition to either an 80:20 or a 93:7 E/Z isomer ratio of coordinated imine complex 2a occurs with high diastereoselectivity. The high diastereoselectivity regardless of the reagent E/Z ratio is explained by a difference between the relative kinetic acidities of the E and Z isomers of the imine complex. The kinetic acidities of related isopropylimine and 3-pentylimine complexes were used in explaining these results. PMID- 14531683 TI - Nickel-catalyzed highly regio- and stereoselective three-component assembly of allenes, aryl iodides, and alkenylzirconium reagents. AB - Cyclohexylallene (1a) and alkenylzirconium reagent (n-PrCH=CHZr(Cp)2Cl) undergo three-component assembly with a variety of aromatic iodides (R1-I: R1 = 4-PhOMe, 4-PhCOMe, 4-PhMe, 2-thiophene, 2-PhOMe, 3-PhOMe, and 4-PhCO2Et) in the presence of NiCl2(PPh3)2 and zinc powder in THF at 50 degrees C to afford corresponding three-component assembly products in moderate to good yields with very high regio and stereoselectivity. In addition to cyclohexylallene, also phenylallene, n butylallene, cyclopentylallene, and cycloheptylallene underwent the three component assembling reaction to furnish the corresponding 1,4-diene derivatives in good yields. Several alkenylzirconium reagents (R2CH=CHZr(Cp)2Cl: R2 = Ph (3b), t-Bu, TMS) were also employed for this three-component assembly. The nickel catalyzed three-component assembly was further extended to vinyl iodides. Thus, 1a reacts with 3b and ethyl (Z)-3-iodoacrylate to produce a highly substituted triene in excellent yield. In all of these three-component assembly reactions, the E isomers were formed predominantly with E/Z ratios between 94/6 and 99/1. A possible mechanism for this highly regio- and stereoselective three-component assembly is proposed. PMID- 14531682 TI - Rapid two-step synthesis of mitrin from heparosan: a replacement for heparin. AB - We have engineered a two-step enzymatic synthesis of Mitrin which is a more potent and homogeneous anticoagulant than the current animal-derived heparin. This engineered heparin may have advantages of being free from animal-derived pathogens and may also have reduced side effects such as heparin induced thrombocytopenia. This approach can also be extended to tailor heparin-based drugs with improved therapeutic characteristics to treat other disorders or infections in which heparin-like molecules play a major role. PMID- 14531684 TI - Conical intersections of three electronic states affect the ground state of radical species with little or no symmetry: pyrazolyl. AB - Conical intersections of three states of the same symmetry are located in the pyrazolyl radical using wave functions of approximately 40 million configuration state functions. The three-state intersection is found to be only approximately 3400 cm-1 above the minimum energy structure on the ground electronic state. It is suggested that many organic radicals may also exhibit this feature. PMID- 14531686 TI - Probing the kinetic landscape of transient peptide-protein interactions by use of peptide (15)n NMR relaxation dispersion spectroscopy: binding of an antithrombin peptide to human prothrombin. AB - Protein-ligand interactions may lead to the formation of multiple molecular complexes in dynamic exchange, affecting the kinetic and thermodynamic characteristics of the binding equilibrium. We followed the dissociation kinetics of the transient and specific complex of an antithrombotic peptide N-acetyl Asp(55)-Phe-Glu-Glu-Ile-Pro(60)-Glu-Glu-Tyr-Leu-Gln(65) with human prothrombin by use of (15)N NMR relaxation dispersion spectroscopy of the peptide. Every one of the five (15)N-labeled adjacent residues of the peptide exhibited apparently different kinetic exchange and relaxation behaviors, which were especially evident at different concentrations of prothrombin. Binding-induced (15)N relaxation dispersion of residues Phe(56), Glu(57), Glu(58), and Ile(59) can be fitted phenomenologically to a two-site on-and-off exchange mechanism with physically feasible relaxation and kinetic parameters obtained for residues Phe(56), Glu(58), and Ile(59), independent of the prothrombin concentration. The apparent kinetic parameters of Glu(57) show some dependence on the concentration of prothrombin and the extracted transverse relaxation rate for Glu(57) in the bound state was severalfold higher than that expected for a protein-peptide complex with a size of approximately 72 kDa. In addition, the equilibrium population of the bound peptide obtained for Glu(57) was inconsistent with those for Phe(56), Glu(58), and Ile(59) and with the prothrombin/peptide ratios used in the experiments. These discrepancies can be explained by the presence of two conformations for the peptide-protein complex exchanging at a rate of approximately 100 s(-)(1). In all, our study shows that fast dissociation of protein-peptide complexes can be studied quantitatively using peptide (15)N NMR relaxation dispersion measurements without a precise knowledge of the peptide and protein concentrations. In addition, protein titration was found to improve the accuracy of quantitative analysis and may make it possible to determine the rate of conformational changes within the protein-peptide complex. PMID- 14531685 TI - Extended pi-conjugated dendrimers based on truxene. AB - The largest pi-conjugated dendrimers containing up to nine 10,15-dihydro-5H diindeno[1,2-alpha;1',2';-c]fluorene (truxene) moieties have been prepared with good yields by repetitive Friedel-Crafts acetylation and acid-promoted cyclotrimerization reactions. An alternative approach to the convergent synthesis of desired dendrimers has been developed, in which the core is generated "in situ" by acid-promoted cyclotrimerizations of aryl methyl ketones. This proves valuable to afford large-size and precisely well-defined dendrimers in an accelerated dendrimer-growth strategy utilizing enlarged repeat units. The increasing amount of SiCl4 dramatically improves the yield of cyclotrimerization reactions. The introduction of hexahexyl groups onto C-5, C-10, C-15 positions of the truxene moiety greatly enhances the solubility of our compounds. Further investigation indicates that the torsion angle between the truxene segment and the benzene ring might play a key role in determining the photo properties of pi conjugated dendrimers. PMID- 14531687 TI - The kinetics of helix unfolding of an azobenzene cross-linked peptide probed by nanosecond time-resolved optical rotatory dispersion. AB - The unfolding dynamics of a 16 amino acid peptide (Ac-EACAREAAAREAACRQ-NH(2), FK 11-X) was followed using nanosecond time-resolved optical rotatory dispersion (ORD). The peptide was coupled to an azobenzene linker that undergoes subnanosecond photoisomerization and reisomerizes on a time scale of minutes. When the linker is in the trans form, the peptide favors a more helical structure (66% helix/34% disordered) and when in the cis configuration the helical content is reduced. Unfolding of FK-11-X was rapidly triggered by a 7-ns laser pulse at 355 nm, forming cis azobenzene-linked peptides that maintained the secondary structure (helical or disordered) of their trans azobenzene counterparts. The incompatibility of the instantaneous cis photoproduct with helical secondary structure drives the subsequent peptide unfolding to a new conformational equilibrium between cis helix and cis disordered structures. The kinetic results show a approximately 40% decrease in the time-dependent ORD signal at 230 nm that is best fit to a single-exponential decay with a time constant of 55 +/- 6 ns. Folding and unfolding rates for cis FK-11-X are estimated to be approximately 3.0 x 10(6) s(-)(1) (1/330 ns) and approximately 1.5 x 10(7) s(-)(1) (1/66 ns), respectively. PMID- 14531688 TI - Nanosecond photoreduction of cytochrome p450cam by channel-specific Ru-diimine electron tunneling wires. AB - We report the synthesis and characterization of Ru-diimine complexes designed to bind to cytochrome p450cam (CYP101). The sensitizer core has the structure [Ru(L(2))L'](2+), where L' is a perfluorinated biphenyl bridge (F(8)bp) connecting 4,4'-dimethylbipyridine to an enzyme substrate (adamantane, F(8)bp Ad), a heme ligand (imidazole, F(8)bp-Im), or F (F(9)bp). The electron-transfer (ET) driving force (-deltaG degrees ) is varied by replacing the ancillary 2,2' bipyridine ligands with 4,4',5,5'-tetramethylbipyridine (tmRu). The four complexes all bind p450cam tightly: Ru-F(8)bp-Ad (1, K(d) = 0.077 microM); Ru F(8)bp-Im (2, K(d) = 3.7 microM); tmRu-F(9)bp (3, K(d) = 2.1 microM); and tmRu F(8)bp-Im (4, K(d) = 0.48 microM). Binding is predominantly driven by hydrophobic interactions between the Ru-diimine wires and the substrate access channel. With Ru-F(8)bp wires, redox reactions can be triggered on the nanosecond time scale. Ru-wire 2, which ligates the heme iron, shows a small amount of transient heme photoreduction (ca. 30%), whereas the transient photoreduction yield for 4 is 76%. Forward ET with 4 occurs in roughly 40 ns (k(f) = 2.8 x 10(7) s(-)(1)), and back ET (Fe(II) --> Ru(III), k(b) approximately 1.7 x 10(8) s(-)(1)) is near the coupling-limited rate (k(max)). Direct photoreduction was not observed for 1 or 3. The large variation in ET rates among the Ru-diimine:p450 conjugates strongly supports a through-bond model of Ru-heme electronic coupling. PMID- 14531689 TI - Electron transfer reactions of redox cofactors in spinach photosystem I reaction center protein in lipid films on electrodes. AB - Thin film voltammetry was used to obtain direct, reversible, electron transfer between electrodes and spinach Photosystem I reaction center (PS I) in lipid films for the first time. This reaction center (RC) protein retains its native conformation in the films, and AFM showed that film structure rearranges during the first several minutes of rehydration of the film. Two well-defined chemically reversible reduction-oxidation peaks were observed for native PS I in the dimyristoylphosphatidylcholine films, and were assigned to phylloquinone, A(1) (E(m) = -0.54 V) and iron-sulfur clusters, F(A)/F(B) (E(m) = -0.19 V) by comparisons with PS I samples selectively depleted of these cofactors. Observed E(m) values may be influenced by protein-lipid interactions and electrode double layer effects. Voltammetry was consistent with simple kinetically limited electron transfers, and analysis of reduction-oxidation peak separations gave electrochemical rate constants of 7.2 s(-)(1) for A(1) and 65 s(-)(1) for F(A)/F(B). A catalytic process was observed in which electrons were injected from PS I in films to ferredoxin in solution, mimicking in vivo electron shuttle from the terminal F(A)/F(B) cofactors to soluble ferredoxin during photosynthesis. PMID- 14531690 TI - Microcin J25 has a threaded sidechain-to-backbone ring structure and not a head to-tail cyclized backbone. AB - Microcin J25 is a 21 amino acid bacterial peptide that has potent antibacterial activity against Gram-negative bacteria, resulting from its interaction with RNA polymerase. The peptide was previously proposed to have a head-to-tail cyclized peptide backbone and a tight globular structure (Blond, A., Peduzzi, J., Goulard, C., Chiuchiolo, M. J., Barthelemy, M., Prigent, Y., Salomon, R. A., Farias, R. N., Moreno, F. & Rebuffat, S. Eur. J. Biochem. 1999, 259, 747-755). It exhibits remarkable thermal stability for a peptide of its size lacking disulfide bonds and in part this was previously proposed to derive from its macrocyclic structure. We show here that in fact the peptide does not have a head-to-tail cyclic structure but rather a side chain to backbone cyclization between Glu8 and the N-terminus. This creates an embedded ring that is threaded by the C-terminal tail of the molecule, forming a noose-like feature. The three-dimensional structure deduced from NMR data suggests that slippage of the noose is prevented by two aromatic residues flanking the embedded ring. Unthreading does not occur even when the molecule is enzymatically digested with thermolysin. The new structural interpretation fully accounts for previously reported NMR and biophysical data and is consistent with the remarkable stability of this potent antimicrobial peptide. PMID- 14531692 TI - Rapid sample-mixing technique for transient NMR and photo-CIDNP spectroscopy: applications to real-time protein folding. AB - We describe the development and application of a novel rapid sample-mixing technique for real-time NMR (nuclear magnetic resonance) spectroscopy. The apparatus consists of an insert inside a conventional NMR tube coupled to a rapid injection syringe outside the NMR magnet. Efficient and homogeneous mixing of solutions in the NMR tube is achieved with a dead time of tens of milliseconds, without modification of the NMR probe or additional hardware inside the magnet. Provision is made for the inclusion of an optical fiber to allow in situ laser irradiation of samples, for example to generate photo-CIDNP (chemically induced dynamic nuclear polarization). An NMR water suppression method has been implemented to allow experiments in H(2)O as well as in deuterated solvents. The performance of the device has been tested and optimized by a variety of methods, including sensitive detection of residual pH gradients and the use of NMR imaging to monitor the extent of mixing in real time. The potential utility of this device, in conjunction with the sensitivity and selectivity of photo-CIDNP, is demonstrated by experiments on the protein hen lysozyme. These measurements involve the direct detection of spectra during real-time refolding, and the use of CIDNP pulse labeling to study a partially unfolded state of the protein under equilibrium conditions. Magnetization transfer from this disordered state to the well-characterized native state provides evidence for the remarkable persistence of nativelike elements of structure under conditions in which the protein is partially denatured and aggregation prone. PMID- 14531691 TI - Structure of microcin J25, a peptide inhibitor of bacterial RNA polymerase, is a lassoed tail. AB - Microcin J25 (MccJ25) is a 21-amino acid peptide inhibitor active against the DNA dependent RNA polymerase of Gram negative bacteria. Previously, the structure of MccJ25 was reported to be a head-to-tail circle, cyclo( G(1)GAGHVPEYF(10)VGIGTPISFY(20)G-). On the basis of biochemical studies, mass spectrometry, and NMR, we show that this structure is incorrect, and that the peptide has an extraordinary structural fold. MccJ25 contains an internal lactam linkage between the alpha-amino group of Gly1 and the gamma-carboxyl of Glu8. The tail (Tyr9-Gly21) passes through the ring (Gly1-Glu8), with Phe19 and Tyr20 straddling each side of the ring, sterically trapping the tail in a noncovalent interaction we call a lassoed tail. PMID- 14531693 TI - Intervalence near-IR spectra of delocalized dinitroaromatic radical anions. AB - The Class III (delocalized) intervalence radical anions of 1,4-dinitrobenzene, 2,6-dinitronaphthalene, 2,6-dinitroanthracene, 9,9-dimethyl-2,7-dinitrofluorene, 4,4'-dinitrobiphenyl, and 1,5-dinitronaphthalene show charge-transfer bands in their near-IR spectra. The dinitroaromatic radical anions have comparable but slightly larger electronic interactions (H(ab) values) through the same aromatic bridges as do the corresponding dianisylamino-substituted radical cations. H(ab) values range from 5410 cm(-)(1) (1,4- dinitrobenzene) to 3400 cm(-)(1) (9,9 dimethyl-2,7-dinitrofluorene), decreasing as the number of bonds between the nitro groups increases, except for the 1,5-dinitronaphthalene radical-anion, which has a coupling similar to that of 9,9-dimethyl-2,7-dinitrofluorene. All charge-transfer bands show vibrational fine structure. The vertical excitation energies (lambda(v)) were estimated from the vibrational components, obtained by simulation of the entire band. The large 2H(ab)/lambda(v) values confirm these radicals to be Class III delocalized mixed-valence species. Analysis using Cave and Newton's generalized Mulliken-Hush theory relating the transition dipole moment to the distance on the diabatic surfaces suggests that the electron transfer distance on the diabatic surfaces, d(ab), is only 26-40% of the nitrogen to-nitrogen distance, which implies that something may be wrong with our analysis. PMID- 14531694 TI - Chiral Ru-based complexes for asymmetric olefin metathesis: enhancement of catalyst activity through steric and electronic modifications. AB - Design, synthesis, characterization, and catalytic activity of six enantiomerically pure Ru-based metathesis catalysts are disclosed (3a-3f). The new chiral catalysts were prepared through steric and electronic alterations of the parent catalyst system (3). The present studies indicate that the effect of structural modifications of chiral complex 3 does not always correspond to those of the related achiral complexes. The present findings illustrate that modified Ru complexes (3e and 3f) deliver reactivity levels that are more than 2 orders of magnitude higher than 3. Reactivity and physical data are provided that shed light on the origin of activity differences. Some members of the new generation of chiral Ru catalysts promote asymmetric ring-opening (AROM) and ring-closing (ARCM) metatheses that cannot be effected by the first generation chiral catalyst (3). PMID- 14531695 TI - Excited-state singlet manifold and oscillatory features of a nonatetraeniminium retinal chromophore model. AB - In this paper we use ab initio multireference Moller-Plesset second-order perturbation theory computations to map the first five singlet states (S(0), S(1), S(2), S(3), and S(4)) along the initial part of the photoisomerization coordinate for the isolated rhodopsin chromophore model 4-cis-gamma-methylnona 2,4,6,8-tetraeniminium cation. We show that this information not only provides an explanation for the spectral features associated to the chromophore in solution but also, subject to a tentative hypothesis on the effect of the protein cavity, may be employed to explain/assign the ultrafast near-IR excited-state absorption, stimulated emission, and transient excited-state absorption bands observed in rhodopsin proteins (e.g. rhodopsin and bacteriorhodopsin). We also show that the results of vibrational frequency computations reveal a general structure for the first (S(1)) excited-state energy surface of PSBs that is consistent with the existence of the coherent oscillatory motions observed both in solution and in bacteriorhodopsin. PMID- 14531696 TI - Theoretical analysis of residual dipolar coupling patterns in regular secondary structures of proteins. AB - A new approach to the interpretation of residual dipolar couplings for the regular secondary structures of proteins is presented. This paper deals with the analysis of the steric and chiral requirements of protein secondary structures and establishes a quantitative correlation between structure periodicity and the experimental values of the backbone residual dipolar couplings. Building on the recent interpretation of the periodicity of residual dipolar couplings in alpha helices (i.e., "dipolar waves"), a general parametric equation for fitting the residual dipolar couplings of any regular secondary structure is derived. This equation interprets the modulation of the residual dipolar couplings' periodicity in terms of the secondary structure orientation with respect to an arbitrary reference frame, laying the groundwork for using backbone residual dipolar couplings as a fast tool for determining protein folding by NMR spectroscopy. PMID- 14531697 TI - Palladium-catalyzed negishi cross-coupling reactions of unactivated alkyl iodides, bromides, chlorides, and tosylates. AB - A single method (2% Pd(2)(dba)(3)/8% PCyp(3)/NMI in THF/NMP at 80 degrees C; Cyp = cyclopentyl) achieves the cross-coupling of a range of beta-hydrogen-containing primary alkyl iodides, bromides, chlorides, and tosylates with an array of alkyl , alkenyl-, and arylzinc halides. The process is compatible with a variety of functional groups, including esters, amides, imides, nitriles, and heterocycles. PMID- 14531698 TI - Nanocrystals modified with peptide nucleic acids (PNAs) for selective self assembly and DNA detection. AB - Gold nanocrystals modified with peptide nucleic acids (PNAs) have been prepared and applied to self-assembly and DNA sensing. Experiments with different PNA structural motifs show that (1). the versatility in PNA synthetic design can be used to modulate the electrostatic surface properties of nanocrystals, presenting an opportunity to control assembly rate and aggregate size, (2). short (6 base) PNAs can hybridize effectively while attached to nanoparticles, providing a route to generating materials with small interparticle spacings, and (3). the superior base pair mismatch selectivity of PNAs is further enhanced on nanosurfaces, enabling PNA-modified nanoparticles to act as highly selective nanoscale sensors, as well as synthons for defect-free self-assembly. This last feature was coupled with a substantial change in colloidal stability upon DNA hybridization to develop a novel colorimetric DNA assay that detects the presence of single base imperfections within minutes. Various modes of PNA hybridization, including the first practical application of PNA-PNA interactions, were used to direct the assembly of nanoparticles into macroscopic arrangements. Shorter duplex interconnects and greater specificity in assembly were obtained compared to similar experiments with DNA-modified nanocrystals. PMID- 14531699 TI - Nonenzymatic breakdown of the tetrahedral (alpha-carboxyketal phosphate) intermediates of MurA and AroA, two carboxyvinyl transferases. Protonation of different functional groups controls the rate and fate of breakdown. AB - The mechanisms of nonenzymatic breakdown of the tetrahedral intermediates (THIs) of the carboxyvinyl transferases MurA and AroA were examined in order to illuminate the interplay between the inherent reactivities of the THIs and the enzymatic strategies used to promote catalysis. THI degradation was through phosphate departure, with C-O bond cleavage. It was acid catalyzed and dependent on the protonation state of the carboxyl of the alpha-carboxyketal phosphate functionality, with ionizations at pK(a) = 3.2 +/- 0.1 and 4.3 +/- 0.1 for MurA and AroA THIs, respectively. The solvent deuterium kinetic isotope effect for MurA THI at pL 2.0 was 1.3 +/- 0.4, consistent with general acid catalysis. The pK(a)'s suggested intramolecular general acid catalysis through protonation of the bridging oxygen of the phosphate, though H(3)O(+) catalysis was also possible. The product distribution varied with pH. The dominant breakdown products were pyruvate + phosphate + R-OH (R-OH = UDP-GlcNAc or shikimate 3 phosphate) at all pH's, particularly low pH. At higher pH's, increasing proportions of ketal, arising from intramolecular substitution of phosphate by the adjacent hydroxyl and the enolpyruvyl products of phosphate elimination were observed. With MurA THI, the product distribution fitted to pK(a)'s 1.6 and 6.2, corresponding to the expected pK(a)'s of a phosphate monoester. C-O bond cleavage was demonstrated by the lack of monomethyl [(33)P]phosphate formed upon degrading MurA [(33)P]THI in 50% methanol. General acid catalysis through the bridging oxygen is consistent with the location of the previously proposed general acid catalyst for THI breakdown in AroA, Lys22. PMID- 14531700 TI - Substrate recognition and channeling of monomodules from the pikromycin polyketide synthase. AB - The unique ability of the pikromycin (Pik) polyketide synthase to generate 12- and 14-membered ring macrolactones presents an opportunity to explore the fundamental processes underlying polyketide synthesis, specifically the mechanistic details of the chain extension process. We have overexpressed and purified PikAIII (module 5) and PikAIV (module 6) and assessed the ability of these proteins to generate tri- and tetraketide lactone products using N acetylcysteamine-activated diketides and (14)C-methylmalonyl-CoA as substrates. Comparison of the stereochemical specificities for PikAIII and PikAIV and the reported values for the DEBS modules reveals significant differences between these systems. PMID- 14531701 TI - Dendrimeric organochalcogen catalysts for the activation of hydrogen peroxide: origins of the "dendrimer effect" with catalysts terminating in phenylseleno groups. AB - Several scenarios were evaluated to explain the large "dendrimer effect" observed in the bromination of cyclohexene with H(2)O(2) and NaBr catalyzed by the addition of Frechet-type dendrimers terminating in -O(CH(2))(3)SePh groups. Although phenylseleninic acid was an efficient catalyst for the oxidation of NaBr with H(2)O(2), first-order rate constants for the selenoxide elimination were too small to produce PhSeO(2)H at a rate sufficient to explain the rates of catalysis and no dendrimer effect was observed in the rates of selenoxide elimination. An induction period was observed using 1-SePh as a catalyst for the oxidation of Br( ) with H(2)O(2). The addition of preformed selenoxide 1-Se(=O)Ph gave immediate catalysis with no induction period. However, rates of oxidation of the selenides with H(2)O(2) under homogeneous or biphasic conditions or with t-BuOOH under homogeneous conditions were too slow to account for the rates of catalysis, and no dendrimer effect was observed in the rates of oxidation. The primary oxidant for converting selenides to selenoxides was "Br(+)" produced initially by the uncatalyzed background reaction of H(2)O(2) with NaBr and then produced catalytically following formation of selenoxide groups. Autocatalysis is observed, and the rate of oxidation increases with the number of SePh groups. Autocatalysis is the source of the large dendrimer effect observed with the SePh series of catalysts. PMID- 14531702 TI - Large-scale synthesis of nearly monodisperse CdSe/CdS core/shell nanocrystals using air-stable reagents via successive ion layer adsorption and reaction. AB - Successive ion layer adsorption and reaction (SILAR) originally developed for the deposition of thin films on solid substrates from solution baths is introduced as a technique for the growth of high-quality core/shell nanocrystals of compound semiconductors. The growth of the shell was designed to grow one monolayer at a time by alternating injections of air-stable and inexpensive cationic and anionic precursors into the reaction mixture with core nanocrystals. The principles of SILAR were demonstrated by the CdSe/CdS core/shell model system using its shell thickness-dependent optical spectra as the probes with CdO and elemental S as the precursors. For this reaction system, a relatively high temperature, about 220 240 degrees C, was found to be essential for SILAR to fully occur. The synthesis can be readily performed on a multigram scale. The size distribution of the core/shell nanocrystals was maintained even after five monolayers of CdS shell (equivalent to about 10 times volume increase for a 3.5 nm CdSe nanocrystal) were grown onto the core nanocrystals. The epitaxial growth of the core/shell structures was verified by optical spectroscopy, TEM, XRD, and XPS. The photoluminescence quantum yield (PL QY) of the as-prepared CdSe/CdS core/shell nanocrystals ranged from 20% to 40%, and the PL full-width at half-maximum (fwhm) was maintained between 23 and 26 nm, even for those nanocrystals for which the UV vis and PL peaks red-shifted by about 50 nm from that of the core nanocrystals. Several types of brightening phenomena were observed, some of which can further boost the PL QY of the core/shell nanocrystals. The CdSe/CdS core/shell nanocrystals were found to be superior in comparison to the highly luminescent CdSe plain core nanocrystals. The SILAR technique reported here can also be used for the growth of complex colloidal semiconductor nanostructures, such as quantum shells and colloidal quantum wells. PMID- 14531703 TI - Highly regio- and stereoselective acylboration, acylsilation, and acylstannation of allenes catalyzed by phosphine-free palladium complexes: an efficient route to a new class of 2-acylallylmetal reagents. AB - A new method for the synthesis of substituted 2-acylallylmetal reagents in a highly regio- and stereoselective fashion involving a three-component assembly of allenes, acyl chlorides, and bimetallic reagents (B-B, Si-Si, and Sn-Sn) catalyzed by phosphine-free palladium complexes is described. Treatment of various allenes (CR(2)R(3)=C=CH(2)) with acyl chlorides (R(1)COCl) and bispinacolatodiboron in the presence of PdCl(2)(CH(3)CN)(2) in toluene at 80 degrees C gave 2-acylallylboronates in moderate to good yields. The acylsilation of allenes with acid chlorides and hexamethyldisilane (5) proceeded successfully in the presence of Pd(dba)(2) in CH(3)CN affording the corresponding allylsilanes (CR(2)R(3)=C(COR(1))CH(2)SiMe(3)) in good to moderate yields. Several chloroformates (R(4)OCOCl) also react with 1,1-dimethylallene (2a) and 5 to afford allylsilanes (CR(2)R(3)=C(COOR(4))CH(2)SiMe(3)) in 66-70% yields. Acylstannation of allenes could also be achieved by slow addition of hexabutylditin (10) to the reaction mixture of acyl chloride (or chloroformate) and allene 2a in CH(3)CN in the presence of Pd(dba)(2) at 60 degrees C; the corresponding 2-substituted allylstannanes were isolated in moderate to good yields. The above catalytic reactions are completely regioselective and highly stereoselective. A mechanism is proposed to account for the catalytic reactions and the stereochemistry. PMID- 14531704 TI - Organolathanide-catalyzed regioselective intermolecular hydroamination of alkenes, alkynes, vinylarenes, di- and trivinylarenes, and methylenecyclopropanes. Scope and mechanistic comparison to intramolecular cyclohydroaminations. AB - Organolanthanide complexes of the type Cp'(2)LnCH(SiMe(3))(2) (Cp' = eta(5) Me(5)C(5); Ln = La, Nd, Sm, Lu) and Me(2)SiCp' '(2)LnCH(SiMe(3))(2) (Cp' ' = eta(5)-Me(4)C(5); Ln = Nd, Sm, Lu) serve as efficient precatalysts for the regioselective intermolecular hydroamination of alkynes R'Ctbd1;CMe (R' = SiMe(3), C(6)H(5), Me), alkenes RCH=CH(2) (R = SiMe(3), CH(3)CH(2)CH(2)), butadiene, vinylarenes ArCH=CH(2) (Ar = phenyl, 4-methylbenzene, naphthyl, 4 fluorobenzene, 4-(trifluoromethyl)benzene, 4-methoxybenzene, 4 (dimethylamino)benzene, 4-(methylthio)benzene), di- and trivinylarenes, and methylenecyclopropanes with primary amines R' 'NH(2) (R' ' = n-propyl, n-butyl, isobutyl, phenyl, 4-methylphenyl, 4-(dimethylamino)phenyl) to yield the corresponding amines and imines. For R = SiMe(3), R = CH(2)=CH lanthanide mediated intermolecular hydroamination regioselectively generates the anti Markovnikov addition products (Me(3)SiCH(2)CH(2)NHR' ', (E)-CH(3)CH=CHCH(2)NHR' '). However, for R = CH(3)CH(2)CH(2), the Markovnikov addition product is observed (CH(3)CH(2)CH(2)CHNHR' 'CH(3)). For internal alkynes, it appears that these regioselective transformations occur under significant stereoelectronic control, and for R' = SiMe(3), rearrangement of the product enamines occurs via tautomerization to imines, followed by a 1,3-trimethylsilyl group shift to stable N-SiMe(3)-bonded CH(2)=CMeN(SiMe(3))R' ' structures. For vinylarenes, intermolecular hydroamination with n-propylamine affords the anti-Markovnikov addition product beta-phenylethylamine. In addition, hydroamination of divinylarenes provides a concise synthesis of tetrahydroisoquinoline structures via coupled intermolecular hydroamination/subsequent intramolecular cyclohydroamination sequences. Intermolecular hydroamination of methylenecyclopropane proceeds via highly regioselective exo-methylene C=C insertion into Ln-N bonds, followed by regioselective cyclopropane ring opening to afford the corresponding imine. For the Me(2)SiCp' '(2)Nd-catalyzed reaction of Me(3)SiCtbd1;CMe and H(2)NCH(2)CH(2)CH(2)CH(3), DeltaH() = 17.2 (1.1) kcal mol(-)(1) and DeltaS() = -25.9 (9.7) eu, while the reaction kinetics are zero order in [amine] and first-order in both [catalyst] and [alkyne]. For the same substrate pair, catalytic turnover frequencies under identical conditions decrease in the order Me(2)SiCp' '(2)NdCH(SiMe(3))(2) > Me(2)SiCp' '(2)SmCH(SiMe(3))(2) > Me(2)SiCp' '(2)LuCH(SiMe(3))(2) > Cp'(2)SmCH(SiMe(3))(2), in accord with documented steric requirements for the insertion of olefinic functionalities into lanthanide-alkyl and -heteroatom sigma-bonds. Kinetic and mechanistic evidence argues that the turnover-limiting step is intermolecular C=C/Ctbd1;C bond insertion into the Ln-N bond followed by rapid protonolysis of the resulting Ln-C bond. PMID- 14531705 TI - Crystal environments probed by EPR spectroscopy. Variations in the EPR spectra of Co(II)(octaethylporphyrin) doped in crystalline diamagnetic hosts and a reassessment of the electronic structure of four-coordinate cobalt(II). AB - The powder and single-crystal EPR spectra of Co(II)(OEP) (OEP is the dianion of octaethylporphyrin) doped into a range of diamagnetic crystals including simple four-coordinate hosts, H(2)(OEP), the triclinic B form of Ni(II)(OEP), the tetragonal form of Ni(II)(OEP) and Zn(II)(OEP); five-coordinate hosts, micro dioxane)[Zn(II)(OEP)](2) and (py)Zn(II)(OEP); six-coordinate hosts, (py)(2)Zn(II)(OEP) and (py)(2)Mg(II)(OEP); and hosts containing fullerenes, C(60).2Zn(II)(OEP).CHCl(3), C(70).Ni(II)(OEP).C(6)H(6).CHCl(3), and C(60).Ni(II)(OEP).2C(6)H(6) have been obtained and analyzed. Spectra were simulated using a program that employed the exact diagonalization of the 16 x 16 complex spin Hamiltonian matrix. The EPR spectra of these doped samples are very sensitive to the environment within each crystal with the crystallographic site symmetry determining whether axial or rhombic resonance patterns are observed. For Co(II)(OEP) doped into tetragonal Ni(II)(OEP) (which displays a very large g( perpendicular ) of 3.405 and a very small g( parallel ) of 1.544) and several other crystals containing four-coordinate metal sites, the g components could not be fit using existing theory with the assumption of the usual z(2) ground state. However, reasonable agreement of the observed EPR parameters could be obtained by assuming that the unpaired electron resides in an xy orbital in the four coordinate complexes. PMID- 14531706 TI - Electronic structure of linear thiophenolate-bridged heteronuclear complexes [LFeMFeL](n)(+) (M = Cr, Co, Fe; n = 1-3): a combination of kinetic exchange interaction and electron delocalization. AB - The electronic properties of the isostructural series of heterotrinuclear thiophenolate-bridged complexes of the general formula [LFeMFeL](n)(+) with M = Cr, Co and Fe where L represents the trianionic form of the ligand 1,4,7-tris(4 tertbutyl-2-mercaptobenzyl)-1,4,7-triazacyclononane, synthesized and investigated by a number of experimental techniques in the previous work(1), are subjected now to a theoretical analysis. The low-lying electronic excitations in these compounds are described within a minimal model supported by experiment and quantum chemistry calculations. It was found indeed that various experimental data concerning the magnetism and electron delocalization in the lowest states of all seven compounds are completely reproduced within a model which includes the electron transfer between magnetic orbitals at different metal centers and the electron repulsion in these orbitals (the Hubbard model). Moreover, due to the trigonal symmetry of the complexes, only the electron transfer between nondegenerate orbital, a(1), originating from the t(2g) shell of each metal ion in a pseudo-octahedral coordination, is relevant for the lowest states. An essential feature resulting from quantum chemistry calculations, allowing to explain the unusual magnetic properties of these compounds, is the surprisingly large value and, especially, the negative sign of the electron transfer between terminal iron ions, beta'. According to their electronic properties the series of complexes can be divided as follows: (1). The complexes [LFeFeFeL](3+) and [LFeCrFeL](3+) show localized valences in the ground electronic configuration. The strong antiferromagnetic exchange interaction and the resulting spin 1/2 of the ground-state arise from large values of the transfer parameters. (2). In the complex [LFeCrFeL](+), due to a higher energy of the magnetic orbital on the central Cr ion than on the terminal Fe ones, the spin 3/2 and the single unpaired a(1) electron are almost localized at the chromium center in the ground state. (3). The complex [LFeCoFeL](3+) has one ground electronic configuration in which two unpaired electrons are localized at terminal iron ions. The ground-state spin S = 1 arises from a kinetic mechanism involving the electron transfer between terminal iron ions as one of the steps. Such a mechanism, leading to a strong ferromagnetic interaction between distant spins, apparently has not been discussed before. (4). The complex [LFeFeFeL](2+) is characterized by both spin and charge degrees of freedom in the ground manifold. The stabilization of the total spin zero or one of the itinerant electrons depends on beta', i.e., corresponds to the observed S = 1 for its negative sign. This behavior does not fit into the double exchange model. (5). In [LFeCrFeL](2+) the delocalization of two itinerant holes in a(1) orbitals takes place over the magnetic core of chromium ion. Although the origin of the ground-state spin S = 2 is the spin dependent delocalization, the spectrum of the low-lying electronic states is again not of a double exchange type. (6). Finally, the complex [LFeCoFeL](2+) has the ground configuration corresponding to the electron delocalization between terminal iron atoms. The estimated magnitude of the corresponding electron transfer is smaller than the relaxation energy of the nuclear distortions induced by the electron localization at one of the centers, leading to vibronic valence trapping observed in this compound. PMID- 14531707 TI - Synthesis, cyclic voltammetric studies, and electrogenerated chemiluminescence of a new phenylquinoline-biphenothiazine donor-acceptor molecule. AB - We report the synthesis, electrochemistry, and luminescence of a novel ECL emitting compound containing two electron-accepting hexyl-phenylquinoline groups covalently attached to the 3,3'-positions of the electron-donating 10,10' dimethylbiphenothiazine group. The optimized geometry as determined from semiempirical MNDO calculations shows that the two quinoline groups are twisted 82.5 degrees from the two phenothiazine rings, indicating a lack of electron delocalization among these groups. This unique geometry allows generation of localized radical cations and radical anions capable of generating ECL upon annihilation. However, the phenothiazine rings are twisted 46.5 degrees relative to each other, suggesting possible interactions between the two moieties. This is evident in the electrochemical behavior in which two closely spaced one-electron oxidations, rather than a single two-electron oxidation wave, were observed. The photophysical properties of BHQ-BPZ show strong resemblances to the parent compound, BPQ-PTZ, which contains a single phenothiazine moiety. In addition, the ECL spectrum produced via radical ion annihilation shows good agreement with the fluorescence emission of the compound. PMID- 14531708 TI - Analysis of proton-proton transfer dynamics in rotating solids and their use for 3D structure determination. AB - A detailed analysis of proton-proton-transfer dynamics under magic angle spinning NMR is presented. Results obtained on model compounds are evaluated under different experimental conditions and NMR mixing schemes. It is shown that the resulting buildup rates can be interpreted in terms of internuclear proton-proton distances provided that an appropriate theoretical description is chosen. As demonstrated in two test applications, these dependencies can be used in the context of a three-dimensional structure determination in the solid state. PMID- 14531709 TI - Single-molecule spectroscopy of interfacial electron transfer. AB - It is widely appreciated that single-molecule spectroscopy (SMS) can be used to measure properties of individual molecules which would normally be obscured in an ensemble-averaged measurement. In this report we show how SMS can be used to measure photoinduced interfacial electron transfer (IET) and back electron transfer rates in a prototypical chromophore-bridge-electrode nonadiabatic electron transfer system. N-(1-hexylheptyl)-N'-(12-carboxylicdodecyl)perylene 3,4,9,10-tetracarboxylbisimide was synthesized and incorporated into mixed self assembled monolayers (SAMs) on an ITO (tin-doped indium oxide, a p-type semiconductor) electrode. Single-molecule fluorescence time trajectories from this system reveals "blinks", momentary losses in fluorescence (>20 ms to seconds in duration), which are attributed to discrete electron transfer events: electron injection from the perylene chromophore into the conduction band of the ITO leads to the loss of fluorescence, and charge recombination (back electron transfer) leads to the return of fluorescence. Such blinks are not observed when an electrode is not present. The fluorescence trajectories were analyzed to obtain the forward and back electron rates; the measured rates are found to lie in the millisecond to second regime. Different rates are observed for different molecules, but the lifetime distributions for the forward or back electron transfer for any given molecule are well fit by single exponential kinetics. The methodology used is applicable to a wide variety of systems and can be used to study the effects of distance, orientation, linker, environment, etc. on electron transfer rates. The results and methodology have implications for molecular electronics, where understanding and controlling the range of possible behaviors inherent to molecular systems will likely be as important as understanding the individual behavior of any given molecule. PMID- 14531710 TI - Oxidation mechanism of aromatic peroxy and bicyclic radicals from OH-toluene reactions. AB - Theoretical calculations have been performed to investigate mechanistic features of OH-initiated oxidation reactions of toluene. Aromatic peroxy radicals arising from initial OH and subsequent O(2) additions to the toluene ring are shown to cyclize to form bicyclic radicals rather than undergoing reaction with NO under atmospheric conditions. Isomerization of bicyclic radicals to more stable epoxide radicals possesses significantly higher barriers and, hence, has slower rates than O(2) addition to form bicyclic peroxy radicals. At each OH attachment site, only one isomeric pathway via the bicyclic peroxy radical is accessible to lead to ring cleavage. The study provides thermochemical and kinetic data for quantitative assessment of the photochemical production potential of ozone and formation of toxic products and secondary organic aerosol from toluene oxidation. PMID- 14531714 TI - Cathode ray tube phosphors. PMID- 14531715 TI - Converting carbon dioxide into carbamato derivatives. PMID- 14531716 TI - Structural changes accompanying intramolecular electron transfer: focus on twisted intramolecular charge-transfer states and structures. PMID- 14531717 TI - Effect of molecular chirality on the morphology of biomimetic langmuir monolayers. PMID- 14531718 TI - Chelation-controlled Bergman cyclization: synthesis and reactivity of enediynyl ligands. PMID- 14531719 TI - Transition-metal-catalyzed reactions in steroid synthesis. PMID- 14531720 TI - The nitroso ene reaction: a regioselective and stereoselective allylic nitrogen functionalization of mechanistic delight and synthetic potential. PMID- 14531721 TI - Dietary effects on drug metabolism and transport. AB - Metabolic food-drug interactions occur when the consumption of a particular food modulates the activity of a drug-metabolising enzyme system, resulting in an alteration of the pharmacokinetics of drugs metabolised by that system. A number of these interactions have been reported. Foods that contain complex mixtures of phytochemicals, such as fruits, vegetables, herbs, spices and teas, have the greatest potential to induce or inhibit the activity of drug-metabolising enzymes, although dietary macroconstituents (i.e. total protein, fat and carbohydrate ratios, and total energy intake) can also have effects. Particularly large interactions may result from the consumption of herbal dietary supplements. Cytochrome P450 (CYP) 3A4 appears to be especially sensitive to dietary effects, as demonstrated by reports of potentially clinically important interactions involving orally administered drugs that are substrates of this enzyme. For example, interactions of grapefruit juice with cyclosporin and felodipine, St John's wort with cyclosporin and indinavir, and red wine with cyclosporin, have the potential to require dosage adjustment to maintain drug concentrations within their therapeutic windows. The susceptibility of CYP3A4 to modulation by food constituents may be related to its high level of expression in the intestine, as well as its broad substrate specificity. Reported ethnic differences in the activity of this enzyme may be partly due to dietary factors. Food-drug interactions involving CYP1A2, CYP2E1, glucuronosyltransferases and glutathione S transferases have also been documented, although most of these interactions are modest in magnitude and clinically relevant only for drugs that have a narrow therapeutic range. Recently, interactions involving drug transporters, including P-glycoprotein and the organic anion transporting polypeptide, have also been identified. Further research is needed to determine the scope, magnitude and clinical importance of food effects on drug metabolism and transport. PMID- 14531722 TI - Macromolecular therapeutics: advantages and prospects with special emphasis on solid tumour targeting. AB - Macromolecular drugs (also referred to as polymeric drugs) are a diverse group of drugs including polymer-conjugated drugs, polymeric micelles, liposomal drugs and solid phase depot formulations of various agents. In this review we will consider only water-soluble macromolecular drugs. In common, such drugs have high molecular weights, more than 40 kDa, which enables them to overcome renal excretion. Consequently, this group of drugs can attain prolonged plasma or local half-lives. The prolonged circulating time of these macromolecules enables them to utilise the vascular abnormalities of solid tumour tissues, a phenomenon called the enhanced permeability and retention (EPR) effect. The EPR effect facilitates extravasation of polymeric drugs more selectively at tumour tissues, and this selective targeting to solid tumour tissues may lead to superior therapeutic benefits with fewer systemic adverse effects. This contrasts with conventional low-molecular-weight drugs, where intratumour concentration diminishes rapidly in parallel with plasma concentration. The EPR effect is also operative in inflammatory tissues, which justifies the development and use of this class of drugs in infectious and inflammatory conditions. At the present time, several polymeric drugs have been approved by regulatory agencies. These include zinostatin stimalamer (copolymer styrene maleic acid-conjugated neocarzinostatin, or SMANCS) and polyethyleneglycol-conjugated interferon-alpha 2a. This article discusses these and other polymeric drugs in the setting of targeting to solid tumours. PMID- 14531724 TI - Clinical pharmacokinetics of linezolid, a novel oxazolidinone antibacterial. AB - Linezolid is the first antibacterial to be approved from the oxazolidinone class. The drug has substantial antimicrobial activity against Gram-positive organisms such as streptococci, staphylococci and enterococci, including species resistant to conventional antibacterial treatment. Linezolid is fully bioavailable following oral administration when compared with intravenous administration. Maximum plasma linezolid concentrations are usually achieved between 1 and 2 hours after oral administration. Food slightly decreases the rate, but not the extent, of absorption. The distribution of linezolid is approximately equivalent to total body water, and there is low protein binding (31%) to serum albumin. The elimination half-life of linezolid is 5-7 hours, and twice-daily administration of 400-600 mg provides steady-state concentrations in the therapeutic range. Linezolid is mainly cleared by non-renal clearance to two metabolites and renal clearance of the parent compound. Approximately 50% of an administered dose appears in the urine as the two major metabolites, and approximately 35% appears as parent drug. A small degree of nonlinearity has been observed, with a 30% decrease in clearance after a 5-fold increase in dose. The nonlinearity is not relevant over the therapeutic dosage range. Plasma linezolid concentrations in elderly patients, patients with mild to moderate hepatic impairment or mild to severe renal impairment are similar to those achieved in young or healthy volunteers. Higher concentrations are observed in women as compared with men, but the difference is not sufficient to warrant an adjustment in dosage. In patients with severe renal impairment requiring haemodialysis, the exposure to the two primary metabolites was 7 to 8-fold higher than in patients with normal renal function. Therefore, linezolid should be used with caution in patients with severe renal insufficiency. A higher clearance of linezolid was found in children as compared with adults, and therefore higher daily dosages per kg bodyweight are required in children. There is no pharmacokinetic interaction when linezolid is coadministered with aztreonam, gentamicin or warfarin. Linezolid is a mild, reversible, inhibitor of monoamine oxidases A and B. Coadministration of linezolid with the adrenergic agents pseudoephedrine and phenylpropanolamine resulted in increases in blood pressure relative to these agents alone or to placebo. The degree of the change in blood pressure was within that associated with normal daily activities. No interaction was observed when linezolid was coadministered with the serotonergic agent dextromethorphan. PMID- 14531723 TI - Absorption enhancers for nasal drug delivery. AB - This paper describes the basic concepts for the transmucosal delivery of drugs, and in particular the use of the nasal route for delivery of challenging drugs such as polar low-molecular-weight drugs and peptides and proteins. Strategies for the exploitation of absorption enhancers for the improvement of nasal delivery are discussed, including consideration of mechanisms of action and the correlation between toxic effect and absorption enhancement. Selected enhancer systems, such as cyclodextrins, phospholipids, bioadhesive powder systems and chitosan, are discussed in detail. Examples of the use of these enhancers in preclinical and clinical studies are given. Methods for assessing irritancy and damage to the nasal membrane from the use of absorption enhancers are also described. Finally, the mucosal use of absorption enhancers (chitosan) for the improved nasal delivery of vaccines is reported with reference to recent phase I/II clinical studies. PMID- 14531726 TI - Forecasting of blood tacrolimus concentrations based on the Bayesian method in adult patients receiving living-donor liver transplantation. AB - OBJECTIVE: To evaluate Bayesian prediction of blood tacrolimus concentrations in adult patients receiving living-donor liver transplantation (LDLT) using previously obtained population pharmacokinetic parameters. PATIENTS AND METHODS: Data were retrospectively collected from 47 adult patients receiving LDLT who were not included in the estimation of population pharmacokinetic parameters. Blood tacrolimus concentrations were predicted without or with the empirical Bayesian method using sparse samples obtained in the previous week. Predictive performance of the concentrations was evaluated by the mean prediction error (ME), mean absolute prediction error (MAE) and root mean square error (RMSE) as well as the percentage of successful predictions (percentage of absolute prediction error less than 3 microg/L, %PRED3). RESULTS: Concentrations predicted by the population mean pharmacokinetic parameter values coincided well with observed concentrations during the period of tacrolimus infusion immediately after the operation. For concentrations during subsequent oral therapy with tacrolimus, predictability by the population mean pharmacokinetic parameter values alone was not satisfactory. Bayesian forecasting using one or two blood concentrations obtained in the previous week significantly decreased (p<0.05) MAE and RMSE compared with predictions based on the population mean pharmacokinetic parameters on postoperative days 21 and 28, but not on day 14. During postoperative days 15-21, %PRED3 was increased to 68.6% or 71.2% with the Bayesian method using one or two blood concentrations, respectively, from 44.9% with the population mean pharmacokinetic parameter values. CONCLUSION: The present study demonstrated the applicability of the Bayesian method with use of one or two samples for prediction of blood tacrolimus concentrations in adult patients receiving LDLT. PMID- 14531727 TI - Polarized cultures of human airway epithelium from nasal scrapings and bronchial brushings. AB - Airway epithelial cultures are generally derived from tracheas postmortem or from surgical specimens of nasal polyps or turbinates. Scrapings of the mucosal surface have been little used as starting material for cultures because of their low yield of epithelial cells and their contamination with mucous secretions, blood, and underlying connective tissue. For the first time, we report that human airway epithelial cells obtained from nasal scrapings or bronchial brushings can be grown in culture to produce polarized cell sheets suitable for studies of vectorial transport. PMID- 14531725 TI - Clinical pharmacokinetics of atorvastatin. AB - Hypercholesterolaemia is a risk factor for the development of atherosclerotic disease. Atorvastatin lowers plasma low-density lipoprotein (LDL) cholesterol levels by inhibition of HMG-CoA reductase. The mean dose-response relationship has been shown to be log-linear for atorvastatin, but plasma concentrations of atorvastatin acid and its metabolites do not correlate with LDL-cholesterol reduction at a given dose. The clinical dosage range for atorvastatin is 10-80 mg/day, and it is given in the acid form. Atorvastatin acid is highly soluble and permeable, and the drug is completely absorbed after oral administration. However, atorvastatin acid is subject to extensive first-pass metabolism in the gut wall as well as in the liver, as oral bioavailability is 14%. The volume of distribution of atorvastatin acid is 381L, and plasma protein binding exceeds 98%. Atorvastatin acid is extensively metabolised in both the gut and liver by oxidation, lactonisation and glucuronidation, and the metabolites are eliminated by biliary secretion and direct secretion from blood to the intestine. In vitro, atorvastatin acid is a substrate for P-glycoprotein, organic anion-transporting polypeptide (OATP) C and H+-monocarboxylic acid cotransporter. The total plasma clearance of atorvastatin acid is 625 mL/min and the half-life is about 7 hours. The renal route is of minor importance (<1%) for the elimination of atorvastatin acid. In vivo, cytochrome P450 (CYP) 3A4 is responsible for the formation of two active metabolites from the acid and the lactone forms of atorvastatin. Atorvastatin acid and its metabolites undergo glucuronidation mediated by uridinediphosphoglucuronyltransferases 1A1 and 1A3. Atorvastatin can be given either in the morning or in the evening. Food decreases the absorption rate of atorvastatin acid after oral administration, as indicated by decreased peak concentration and increased time to peak concentration. Women appear to have a slightly lower plasma exposure to atorvastatin for a given dose. Atorvastatin is subject to metabolism by CYP3A4 and cellular membrane transport by OATP C and P glycoprotein, and drug-drug interactions with potent inhibitors of these systems, such as itraconazole, nelfinavir, ritonavir, cyclosporin, fibrates, erythromycin and grapefruit juice, have been demonstrated. An interaction with gemfibrozil seems to be mediated by inhibition of glucuronidation. A few case studies have reported rhabdomyolysis when the pharmacokinetics of atorvastatin have been affected by interacting drugs. Atorvastatin increases the bioavailability of digoxin, most probably by inhibition of P-glycoprotein, but does not affect the pharmacokinetics of ritonavir, nelfinavir or terfenadine. PMID- 14531728 TI - Significance of conservative asparagine residues in the thermal hysteresis activity of carrot antifreeze protein. AB - The approximately 24-amino-acid leucine-rich tandem repeat motif (PXXXXXLXXLXXLXLSXNXLXGXI) of carrot antifreeze protein comprises most of the processed protein and should contribute at least partly to the ice-binding site. Structural predictions using publicly available online sources indicated that the theoretical three-dimensional model of this plant protein includes a 10-loop beta helix containing the approximately 24-amino-acid tandem repeat. This theoretical model indicated that conservative asparagine residues create putative ice-binding sites with surface complementarity to the 1010 prism plane of ice. We used site specific mutagenesis to test the importance of these residues, and observed a distinct loss of thermal hysteresis activity when conservative asparagines were replaced with valine or glutamine, whereas a large increase in thermal hysteresis was observed when phenylalanine or threonine residues were replaced with asparagine, putatively resulting in the formation of an ice-binding site. These results confirmed that the ice-binding site of carrot antifreeze protein consists of conservative asparagine residues in each beta-loop. We also found that its thermal hysteresis activity is directly correlated with the length of its asparagine-rich binding site, and hence with the size of its ice-binding face. PMID- 14531729 TI - Characterization of a novel binding partner of the melanocortin-4 receptor: attractin-like protein. AB - The gene dosage effect of the MC4-R (melanocortin 4 receptor) on obesity suggests that regulation of MC4-R expression and function is critically important to the central control of energy homoeostasis. In order to identify putative MC4-R regulatory proteins, we performed a yeast two-hybrid screen of a mouse brain cDNA library using the mouse MC4-R intracellular tail (residues 303-332) as bait. We report here on one positive clone that shares 63% amino acid identity with the C terminal part of the mouse attractin gene product, a single-transmembrane-domain protein characterized as being required for agouti signalling through the melanocortin 1 receptor. We confirmed a direct interaction between this ALP (attractin-like protein) and the C-terminus of the mouse MC4-R by glutathione S transferase pulldown experiments, and mapped the regions involved in this interaction using N- and C-terminal truncation constructs; residues 303-313 in MC4-R and residues 1280-1317 in ALP are required for binding. ALP is highly expressed in brain, but also in heart, lung, kidney and liver. Furthermore, co localization analyses in mice showed co-expression of ALP in cells expressing MC4 R in a number of regions known to be important in the regulation of energy homoeostasis by melanocortins, such as the paraventricular nucleus of hypothalamus and the dorsal motor nucleus of the vagus. PMID- 14531730 TI - WBP-2, a WW domain binding protein, interacts with the thyroid-specific transcription factor Pax8. AB - The Pax gene family encodes transcription factors that are essential in organogenesis and in the differentiation of various organs in higher eukaryotes. Pax proteins have a DNA binding domain at the N-terminus, and a transcriptional activation domain at the C-terminus. How these domains interact with the transcriptional machinery of the cell is still unclear. In the present paper, we describe the identification by means of immunological screening of the WW domain binding protein WBP-2 as a biochemical interactor of Pax8 (a WW domain is a protein-interaction domain containing two conserved tryptophan residues). Pax8 is required for the morphogenesis of the thyroid gland and for the maintenance of the thyroid differentiated cellular phenotype. WBP-2 was identified originally as a WW domain binding protein, and its function is still unknown. WBP-2 binds to Pax8 in vitro in pulldown assays, and in vivo in tissue culture cells in co immunoprecipitation assays. Interestingly, Pax8 does not contain a WW domain. Our results point to the identification of a new protein-interacting domain that is present in the C-terminal portion of Pax8 and that is required for protein protein interaction with WBP-2. Our results demonstrate that WBP-2 is not a transcriptional co-activator of Pax8, but rather behaves as an adaptor molecule, as suggested in other studies. PMID- 14531733 TI - Accumulation of manganese superoxide dismutase under metal-depleted conditions: proposed role for zinc ions in cellular redox balance. AB - A diet low in copper results in increased levels of MnSOD (manganese superoxide dismutase), a critical antioxidative enzyme conferring protection against oxidative stress, in rat liver mitochondria. The mechanism for this was investigated using cultured HepG2 cells, a human hepatocellular carcinoma-derived line. MnSOD activity increased 5-7-fold during incubation in a medium supplemented with metal-depleted fetal bovine serum, with a corresponding elevation of its mRNA levels. Metal depletion also decreased CuZnSOD and glutathione peroxidase levels to approx. 70-80% of baseline. When zinc ions were added to the medium at micromolar levels, MnSOD accumulation was suppressed; however, copper ions had essentially no effect on MnSOD expression. Since the intracellular redox status was shifted to a more oxidized state by metal depletion, we examined the DNA-binding activity of NF-kappaB (nuclear factor kappaB), an oxidative stress-sensitive transactivating factor that plays a primary role in MnSOD induction. A gel shift assay indicated that the DNA-binding activity of NF-kappaB was increased in cells maintained in metal-depleted culture, suggesting the involvement of the transactivating function of NF-kappaB in this induction. This was further supported by the observation that curcumin suppressed both the DNA-binding activity of NF-kappaB and the induction of MnSOD mRNA in cells cultivated under metal-depleted conditions. These results suggest that the level of zinc, rather than copper, is a critical regulatory factor in MnSOD expression. It is possible that a deficiency of zinc in the low-copper diet may be primarily involved in MnSOD induction. PMID- 14531732 TI - Regulation of hypoxia-inducible factor-1alpha by nitric oxide through mitochondria-dependent and -independent pathways. AB - Nitric oxide (NO) has been reported both to promote and to inhibit the activity of the transcription factor hypoxia-inducible factor-1 (HIF-1). In order to avoid the pitfalls associated with the use of NO donors, we have developed a human cell line (Tet-iNOS 293) that expresses the inducible NO synthase (iNOS) under the control of a tetracycline-inducible promoter. Using this system to generate finely controlled amounts of NO, we have demonstrated that the stability of the alpha-subunit of HIF-1 is regulated by NO through two separate mechanisms, only one of which is dependent on a functional respiratory chain. HIF-1alpha is unstable in cells maintained at 21% O(2), but is progressively stabilized as the O(2) concentration decreases, resulting in augmented HIF-1 DNA-binding activity. High concentrations of NO (>1 microM) stabilize HIF-1alpha at all O(2) concentrations tested. This effect does not involve the respiratory chain, since it is preserved in cells lacking functional mitochondria (rho(0)-cells) and is not reproduced by other inhibitors of the cytochrome c oxidase. By contrast, lower concentrations of NO (<400 nM) cause a rapid decrease in HIF-1alpha stabilized by exposure of the cells to 3% O(2). This effect of NO is dependent on the inhibition of mitochondrial respiration, since it is mimicked by other inhibitors of mitochondrial respiration, including those not acting at cytochrome c oxidase. We suggest that, although stabilization of HIF-1alpha by high concentrations of NO might have implications in pathophysiological processes, the inhibitory effect of lower NO concentrations is likely to be of physiological relevance. PMID- 14531736 TI - Methodology of the survey: the burden of gastrointestinal diseases in Europe. PMID- 14531731 TI - Bioinformatic analysis of the nucleolus. AB - The nucleolus is a plurifunctional, nuclear organelle, which is responsible for ribosome biogenesis and many other functions in eukaryotes, including RNA processing, viral replication and tumour suppression. Our knowledge of the human nucleolar proteome has been expanded dramatically by the two recent MS studies on isolated nucleoli from HeLa cells [Andersen, Lyon, Fox, Leung, Lam, Steen, Mann and Lamond (2002) Curr. Biol. 12, 1-11; Scherl, Coute, Deon, Calle, Kindbeiter, Sanchez, Greco, Hochstrasser and Diaz (2002) Mol. Biol. Cell 13, 4100-4109]. Nearly 400 proteins were identified within the nucleolar proteome so far in humans. Approx. 12% of the identified proteins were previously shown to be nucleolar in human cells and, as expected, nearly all of the known housekeeping proteins required for ribosome biogenesis were identified in these analyses. Surprisingly, approx. 30% represented either novel or uncharacterized proteins. This review focuses on how to apply the derived knowledge of this newly recognized nucleolar proteome, such as their amino acid/peptide composition and their homologies across species, to explore the function and dynamics of the nucleolus, and suggests ways to identify, in silico, possible functions of the novel/uncharacterized proteins and potential interaction networks within the human nucleolus, or between the nucleolus and other nuclear organelles, by drawing resources from the public domain. PMID- 14531738 TI - Gastro-oesophageal reflux disease and ulcer disease. PMID- 14531737 TI - Gastrointestinal cancers in Europe. PMID- 14531739 TI - The burden of Helicobacter pylori infection in Europe. PMID- 14531740 TI - Coeliac disease in Europe. PMID- 14531741 TI - The burden of gallstone disease in Europe. PMID- 14531742 TI - Liver disease in Europe. PMID- 14531743 TI - Pancreatic diseases. PMID- 14531744 TI - Inflammatory bowel disease. PMID- 14531745 TI - Diverticular disease of the colon in Europe: epidemiology, impact on citizen health and prevention. PMID- 14531746 TI - Functional bowel disorders and irritable bowel syndrome in Europe. PMID- 14531747 TI - Gastrointestinal diseases in the paediatric age groups in Europe: epidemiology and impact on healthcare. PMID- 14531748 TI - Digestive health in the elderly: faecal incontinence in adults. PMID- 14531749 TI - Gastroenterology and hepatology services in Europe. PMID- 14531751 TI - Structural neuroimaging in schizophrenia. PMID- 14531752 TI - Schizophrenia and weight management: a systematic review of interventions to control weight. AB - OBJECTIVE: Weight gain is a frequent side effect of antipsychotic medication which has serious implications for a patient's health and well being. This study systematically reviews the literature on the effectiveness of interventions designed to control weight gain in schizophrenia. METHOD: A systematic search strategy was conducted of major databases in addition to citation searches. Study quality was rated. RESULTS: Sixteen studies met the inclusion criteria. Five of eight pharmacological intervention studies reported small reductions in weight (<5% baseline body weight). All behavioural (including diet and/or exercise) interventions reported small reductions in, or maintenance of, weight. CONCLUSION: Weight loss may be difficult but it is not impossible. Given the inconsistent results, the widespread use of pharmacological interventions cannot be recommended. Both dietary and exercise counselling set within a behavioural modification programme is necessary for sustained weight control. PMID- 14531753 TI - Gray matter features of schizotypal disorder patients exhibiting the schizophrenia-related code types of the Minnesota Multiphasic Personality Inventory. AB - OBJECTIVE: Previous studies have suggested that several code types of the Minnesota Multiphasic Personality Inventory (MMPI) are useful markers for identifying schizophrenia. We hypothesized that schizotypal disorder (STD) patients with such schizophrenia-related code types have the morphological brain abnormalities associated with schizophrenia. METHOD: Voxel-based morphometric analysis with statistical parametric mapping (SPM) 99 software was used to investigate the differences in brain morphology between 14 STD patients with the schizophrenia-related code types of the MMPI and 28 normal individuals. RESULTS: The STD patients showed significantly decreased gray matter volume in the insular regions bilaterally and in the left entorhinal cortex, compared with the controls. CONCLUSION: Our findings suggest that STD patients with the schizophrenia-related code types have volume reductions in these regions as an endophenotype that overlaps with schizophrenia. PMID- 14531754 TI - Trends in mortality from suicide, 1965-99. AB - OBJECTIVE: To analyse trends in mortality from suicide over the period 1965-99. METHOD: Data were derived from the WHO database, including data for 47 countries. RESULTS: In the European Union (EU), all age suicide mortality peaked at 16.1/100,000 in men in 1980-84, and declined thereafter to 14.4/100,000 in 1995 98. In females, the fall was 29% to reach 4.6/100,000. A similar pattern of trends was observed in several eastern European countries. In contrast, mortality from suicide rose substantially in the Russian Federation, from 37.7/100,000 in males in 1985-89 to 58.3/100,000 in 1995-98 (+55%), and to 9.5/100,000 (+12%) in females. In the USA and most other American countries providing data, no consistent pattern was evident for males, but falls were observed in females. Steady declines were registered for Japan, starting from the highest suicide rates worldwide in the late 1950s. Suicide rates were upwards in Ireland, Italy, Spain, the UK, Cuba, Australia and New Zealand. Substantial rises were observed in a few countries (Ireland, Cuba, Mexico, Australia and New Zealand) for young males. CONCLUSION: In spite of mixed trends, suicide remains a significant public health problem worldwide. PMID- 14531755 TI - Attempted suicide among young people: risk factors in a prospective register based study of Danish children born in 1966. AB - OBJECTIVE: In order to prevent suicidal behaviour among adolescents and young adults it would be valuable to know if altering the conditions of their upbringing could reduce their suicidal behaviour. The study surveys possible risk factors. METHOD: Population-based registers covering children born in Denmark in 1966 at the age span of 14-27 years and their parents for: health, education, family dissolution, suicidal behaviour, substance abuse, criminality and unemployment. A discrete-time proportional hazard modelling was used to analyse the longitudinal observations. RESULTS: First-time suicide attempts were associated with parental psychiatric disorder, suicidal behaviour, violence, child abuse and neglect. Increased risks were also found among adolescents and young adults who suffered from psychiatric disorder or physical handicap, had been legally imprisoned, were addicted to drugs, or without graduation, vocational training or employment. CONCLUSION: Stigmatization, social exclusion, and mental disorders in the adolescents or young adults and parents increased risks for attempted suicide. PMID- 14531756 TI - An Australian validation study of the temperament and character inventory. AB - OBJECTIVE: To assess the validity of the temperament and character inventory (TCI) measure in an Australian sample. METHOD: A sample of depressed subjects completed the TCI and a measure assessing personality disorder constructs (PDCs), while family members and psychiatrists also returned PDC ratings. RESULTS: Factor analyses generally supported the TCI constructs, when the temperament and character scales were analysed separately. Self-reported PDC scores were validated against corroborative witness ratings and used to assess the TCIs concurrent validity. Validation analyses supported all TCI temperament scales, but the Self-transcendence Character scale was unassociated with PDC scores. The remaining two character scales (i.e. cooperativeness and self-directedness) were non-specifically associated with all PDC scores. CONCLUSION: Study results support the validity of the TCI scales, but question the best model for conceptualizing the TCI. It is suggested that two character scales quantify disordered functioning, while temperament scales quantify style, constructs that may be inter-related or independent in individual subjects. PMID- 14531757 TI - Properties of the temperament and character inventory in a Chinese sample. AB - OBJECTIVE: To determine the influence of language and culture on the temperament and character (TCI) measure in a Chinese sample. METHOD: We translated the TCI into Mandarin and had a non-psychiatric sample of Malaysian Chinese subjects complete the TCI at baseline and at a 1-month retest, with subsets completing English or Mandarin versions alternatively or on both occasions. Analyses examine the TCI factor structure and any impact of language and culture on TCI scoring. RESULTS: We identified age, gender, occupation and language effects on TCI scale scores. Test-retest reliability was high and not compromised by language. Scale internal consistency was also high. Factor analyses of separate sets of TCI scales corresponded strongly to the structure identified in the TCI development studies. CONCLUSION: The results indicate that TCI is likely to have applicability to Chinese subjects, and argue against properties being constrained by the English language or by western culture. PMID- 14531758 TI - A suggested revision of the Ward Atmosphere Scale. AB - OBJECTIVE: In a previous study, we have suggested a revision of the Anger/Aggression and the Spontaneity subscales. The main aim of this study was to re-evaluate the psychometric properties of the other eight subscales of the Ward Atmosphere Scale. METHOD: A total of 550 patients and 822 staff members on 54 psychiatric wards for psychotic patients completed the WAS and the Good Milieu Index (GMI). We calculated Cronbach's alpha, the Corrected Item Total subscale Correlation, subscale intercorrelations and the correlation between subscales and GMI. RESULTS: By removing a total of 16 items, the psychometric properties improved. The revised subscales had acceptable psychometrics and gave a clearer picture of the relationship between the perceived level of patient satisfaction and the WAS subscale scores. CONCLUSION: The revision suggested in this study 'modernized' several of the subscales. We suggest that this revision is implemented in the future use of the WAS. PMID- 14531759 TI - School achievements and schizophrenia: a case-control study. AB - OBJECTIVE: To compare school performance during middle childhood and adolescence among subjects later diagnosed with schizophrenia and their peers. METHOD: School records were ascertained from archives in 76 acutely symptomatic schizophrenic patients and 146 controls matched for gender and graded by the same teachers. RESULTS: At age 9, no differences in yearly graded school subjects were found. At 12, cases had higher marks in drawing/art and a tendency to excel in language and religion. At 15, the proficient performance in drawing/art remained, but cases performed worse than controls in gymnastics. The cases reported high frequency of school adaptation problems. Severity of psychotic symptoms at admission was associated with lower and declining marks. CONCLUSION: Schizophrenic patients seem to perform as well as their classmates in most subjects taught in compulsory basic school. Findings indicating specific talents in artistic ability and impairments in motor development need to be further investigated. PMID- 14531760 TI - No association between two polymorphisms at the 5HT2A gene and bipolar affective puerperal psychosis. AB - OBJECTIVE: To examine whether variation at two common polymorphisms, T102C and 1438AG, of the serotonin 2A gene (5HT2A) are involved in the puerperal triggering mechanism of bipolar affective puerperal psychosis. METHOD: A total of 242 parous women diagnosed with bipolar disorder were genotyped for the two polymorphisms. Of these, 165 women had experienced a manic or psychotic episode, according to DSM-IV criteria, within 6 weeks of childbirth (the puerperal psychosis group). The comparison group comprised of 77 parous women who had not experienced psychiatric disturbance following childbirth. RESULTS: No significant differences between genotype or allelic frequencies were found between the two groups for either polymorphism. CONCLUSION: The results indicate that variation at two common polymorphisms of the 5HT2A gene does not appear to play a major role in the development of bipolar affective puerperal psychosis. PMID- 14531761 TI - Suicidal feelings run high among mothers in refugee camps: a cross-sectional survey. AB - OBJECTIVE: To study levels of mental distress in a sample of Afghan mothers caring for children in two refugee camps in North West Frontier Province (NWFP) of Pakistan. METHOD: Cross-sectional survey of 297 consecutive mothers with young children, attending primary care centres, using a psychiatric screening instrument, the Self-Reporting Questionnaire (SRQ-20). RESULTS: One hundred and six (36%) of women in the sample screened positive for a common mental disorder. Ninety-six (91%) of those screening positive had had suicidal thoughts in the previous month, and nine (8%) rated suicidal feeling as their topmost concern. CONCLUSION: There is a high prevalence and severity of mental distress in Afghan mothers caring for young children in refugee camps. This may have serious long term effects on the psychological and physical development of their children. PMID- 14531766 TI - Resumative synthesis of contents: it is time for further renoprotection. PMID- 14531762 TI - Psychotic disorder in a case with Hallervorden-Spatz disease. AB - OBJECTIVE: Hallervorden-Spatz disease is a rare autosomal recessive condition, with early onset of predominantly extrapyramidal dysfunction. The symptoms of the disease are dystonia, rigidity, choreoathetosis, pyramidal signs, and intellectual decline. Recent genetic studies mapped the disease to chromosome 20p12.3-p13, and identified mutations in the pantothenate kinase gene. This report describes a childhood onset case of Hallervorden-Spatz disease with schizophreniform psychotic symptoms. Former reports about the psychiatric comorbidity generally included depressive disorder. METHOD: A single case report. RESULTS: A 14-year-old boy with Hallervorden-Spatz disease presented a psychotic episode with prominent auditory hallucinations. Symptoms were relieved after neuroleptic treatment. CONCLUSION: To the authors' knowledge, this is the first published report of the disease with psychotic symptoms. The contribution of basal ganglia, with their wide projections, to the emergence of psychotic symptoms was discussed. PMID- 14531767 TI - Hemoglobin predicts long-term survival in dialysis patients: a 15-year single center longitudinal study and a correlation trend between prealbumin and hemoglobin. AB - BACKGROUND: Dialysis patients have much higher mortality rates than the general population. Anemia is a common complication of uremia and a major contributor to morbidity and mortality in dialysis patients. The benefits of anemia correction using recombinant human erythropoietin (rHuEPO) are well established. Optimum hemoglobin level for dialysis patients remain controversial. We have investigated the association of enrollment hemoglobin with long-term survival in hemodialysis (HD) and peritoneal dialysis (PD) patients. METHODS: We enrolled 529 HD and 326 PD patients from 1987 and followed them to April 2003. Demographics, enrollment, and clinical and laboratory data were recorded. The Kaplan-Meier method was used to compute observed survival, and the multivariate Cox regression analysis was used to identify the independent predictors of mortality risk. RESULTS: Mean ages of HD and PD patients were 60 +/- 16 (SD) and 54 +/- 16 (SD) years, respectively. Forty-seven percent of HD patients and 41% of PD patients were diabetic. Mean enrollment hemoglobin levels of HD and PD patients were 9.44 +/- 1.9 and 9.61 +/- 1.77 g/dL respectively. Cumulative 15 year observed survivals of HD (P = 0.05) and PD (P = 0.032) patients with hemoglobin levels greater or equal to 12 g/dL were higher than those with hemoglobin levels less than 12 g/dL. Hemoglobin <12 g/dL was a better predictor of mortality in nondiabetics than diabetics, particularly in HD patients. Both in HD and PD diabetic patients, hemoglobin was not a significant predictor of mortality. By Cox regression analysis, after adjusting for age, race, gender, and months on dialysis at enrollment, the relative risk of mortality of patients with hemoglobin <12 g/dL was 2.13-fold (P = 0.008) higher for HD and 1.85-fold (P = 0.06) higher for PD compared to those with hemoglobin >/=12 g/dL (P = 0.035). A logistic regression analysis revealed a strong inverse relationship between the hemoglobin level and the odds risk of death in HD (OR = 0.83, P = 0.008) and in PD (OR = 0.85, P = 0.02) patients. CONCLUSION: Enrollment hemoglobin is a predictor of long-term survival in HD and PD patients. Patients with hemoglobin levels that are higher than current treatment recommendations (>12 g/dL) may benefit from long-term survival. Survival of dialysis patients may be improved by better management of malnutrition and anemia. PMID- 14531768 TI - Prospective trials on anemia of chronic disease: the Trial to Reduce Cardiovascular Events with Aranesp Therapy (TREAT). PMID- 14531769 TI - Anemia of chronic disease: past, present, and future. PMID- 14531770 TI - Chronic kidney disease and cardiovascular disease in the Medicare population. AB - BACKGROUND: The extent of diabetes, chronic kidney disease (CKD), and cardiovascular disease (CVD) in the Medicare population is relatively unknown. Also unknown is the effect of these diseases on patient survival before end-stage renal disease (ESRD). METHODS: Prevalent cohorts of Medicare enrollees from 1996 to 2000 were assessed for diabetes and CKD, presence of CVD, and probability of death versus ESRD in the follow-up period. Hospitalization rates and, in diabetics, lipid testing and glycemic control monitoring were also assessed. RESULTS: The prevalence of diabetes in the Medicare population increased at 4.4% per year, reaching 18.9% in the 1999-2000 cohort. Approximately 726,000 elderly Medicare enrollees carry a diagnosis code for CKD. Those with CKD are 5 to 10 times more likely to die before reaching ESRD than the non-CKD group. In CKD patients, CVD is twice as common and advances at twice the rate. Cardiovascular disease advances at a similarly higher rate in CKD patients who die and those who survive to ESRD. Heart failure hospitalizations are 5 times greater in CKD patients and only 30% less than those in dialysis patients. Only half of the CKD patients with diabetes who advance to ESRD had a lipid or glycosylated hemoglobin test done in the year before or after dialysis initiation. CONCLUSION: Diabetes, the leading cause of ESRD, is increasing in the general Medicare population at 4.4% per year. Cardiovascular disease is common, progresses at twice the rate, is associated with death before ESRD, and patients receive suboptimal risk factor monitoring. Active identification and treatment of CKD patients is needed. PMID- 14531771 TI - Anemia as a risk factor for cardiovascular disease. AB - In the present review we examine the physiologic response to chronic anemia and describe potential adverse effects of anemia on myocardial and large arterial remodeling. We present observational data demonstrating that anemia is a risk factor for cardiovascular disease (CVD) outcomes in patients with chronic kidney disease and patients with heart failure. We also present data that have evaluated the relationship of level of hematocrit to CVD outcomes in patients with ischemic heart disease and in patients in the general population. The results from the latter studies have been inconclusive and have been limited by lack of knowledge of the cause of anemia. This is potentially important because iron deficiency anemia may, in fact, improve endothelial function. We conclude that additional randomized controlled trials of treatment of anemia are needed in chronic kidney disease and patients with heart failure; however, further exploration of the cause of anemia and the effect of different types of anemia on various CVD outcomes are needed in patients with ischemic heart disease and patients in the general population. PMID- 14531772 TI - Erythropoietin should be part of congestive heart failure management. AB - BACKGROUND: Up to 64% of patients referred to nephrologists with chronic kidney insufficiency (CKI) have evidence of congestive heart failure (CHF), and most of these patients are also anemic. We have called this triad of anemia, CKI, and CHF the cardio renal anemia (CRA) syndrome. The 3 components of this syndrome form a vicious circle, with each one capable of causing or worsening the other 2. Anemia is found in one-third to one-half of CHF patients and can either cause or worsen the CHF, and can increase the mortality, hospitalization, and malnutrition in this condition. Anemia is also associated with a worsening of renal function in CHF and CKI, causing a more rapid progression to dialysis than is found in those without anemia. Uncontrolled CHF can cause rapid deterioration of renal function and may also cause anemia. Chronic kidney insufficiency can cause anemia and worsen the CHF. METHODS: Aggressive therapy of CHF with all the accepted CHF medications in the accepted doses will often fail to improve the CHF if anemia is also present but is not corrected. However, when the anemia was corrected with subcutaneous erythropoietin and, in some cases, with intravenous iron, the cardiac and patient function and quality of life improved, the need for hospitalization and for high-dose oral and intravenous diuretics was strikingly reduced, and renal function, which had previously been deteriorating, stabilized. RESULTS: Nephrologists should carefully assess the cardiac status of all CKI patients, including routinely getting an echocardiogram and possibly measuring B type natriuretic peptide. Where CHF is present, the indicated CHF agents in the indicated doses should be used. CONCLUSION: Studies show that most cardiologists and internists do not recognize, investigate, or treat the anemia frequently seen in their CHF patients. In our experience cooperation between nephrologists and these specialists has increased their awareness about anemia, resulting in its earlier correction, and thus preventing the deterioration of the CHF, the CKI, and the anemia itself. PMID- 14531773 TI - Effect of erythropoietin on exercise capacity in anemic patients with advanced heart failure. PMID- 14531774 TI - Relationship of bioelectrical impedance parameters to nutrition and survival in peritoneal dialysis patients. AB - BACKGROUND: Malnutrition is highly prevalent in peritoneal dialysis (PD) patients and is associated with higher mortality in these patients. In this study, we have prospectively examined the relationship of bioimpedance indexes to the nutritional status and survival in PD patients. METHODS: We enrolled 48 PD patients beginning in November 2000. On enrollment, bioelectrical impedance analysis (BIA) (BIA-101; RJL/Akern, Clinton Township, MI, USA) was performed and monthly blood was analyzed for biochemical markers, including prealbumin. Patients were followed until April 2003. RESULTS: The mean age of PD patients was 51 +/- 15 (SD) years. Fifty-eight percent of the patients were female and 23% of the patients were diabetic. Mean body mass index (BMI) was 25.7 +/- 5.0 kg/m2. Mean resistance, reactance, and phase angle were 521 +/- 104 ohms, 57 +/- 19 ohms, and 6.16 +/- 1.6 degrees, respectively. During the study period, 8 patients (17%) expired. The Kaplan-Meier method was used to compute observed survival. The cumulative observed survival of PD patients with enrollment phase angle greater than or equal to 6 degrees was significantly higher (P = 0.008) than that of patients with phase angle less than 6. Using Cox's multivariate regression analysis, phase angle was an independent predictor (relative risk = 0.39, P = 0.027) of more than two years' survival in PD patients. Serum prealbumin was directly correlated with phase angle (r = 0.54, P < 0.0001), reactance (r = 0.55, P < 0.0001), and resistance (r = 0.29, P = 0.06). CONCLUSION: BIA indexes reflect nutritional status and may be useful in monitoring nutritional status in PD patients. Phase angle is a strong prognostic index in PD patients. It is useful to incorporate prealbumin and BIA parameters in the regular assessment of PD patients, whose survival may be improved by better management of malnutrition and overall health status. PMID- 14531775 TI - Treating azotemia-induced anemia with erythropoietin improves diabetic eye disease. AB - BACKGROUND: Coincidental with the pandemic growth of diabetes as the prime cause of end-stage renal disease (ESRD), blindness attributable to diabetic retinopathy has become a major concern for all those involved in the care of diabetic ESRD patients. Vision loss is linked to progression of proliferative retinopathy and macular edema. METHODS: Extracted from a study of azotemic anemic pre-ESRD patients treated with erythropoietin, a cohort of five diabetic subjects was reassessed in terms of stability of renal function, changes in blood rheology, and course of diabetic eye disease. RESULTS: All subjects reported subjective improvement in well-being, including enhanced effort tolerance following an increase in hematocrit from a baseline level of to 29.6 +/- 2.0% to a level of 39.5 +/- 2.4% after one year of treatment with erythropoietin (P = <0.0005). Neither hypertension nor deterioration of renal function was noted in any subject. Three patients with macular edema evinced substantive improvement-based stable vision and documented resolution noted in flourescein angiography. CONCLUSION: Erythropoietin treatment of anemic azotemic diabetic patients is well tolerated. In a small observational retrospective study of three patients with macular edema, retention of vision and resolution of exudates was noted. PMID- 14531776 TI - Controversies in iron management. AB - BACKGROUND: Iron therapy is required in hemodialysis patients receiving erythropoietic stimulators in order to achieve the target hemoglobin in the most efficient way. While oral iron has been disappointing in this regard, parenteral iron has been widely used, despite a significant incidence of severe side effects when iron dextran is used. The recent availability of a more effective form of oral iron (heme-iron), and safer forms of parenteral iron (iron sucrose and iron gluconate) has made iron management in this population simpler. Many questions remain, however, about the use, efficacy, and safety of these compounds in hemodialysis patients. METHODS: Current literature was reviewed and combined with the authors' clinical experience to address a number of current questions regarding the use of iron in hemodialysis patients. RESULTS: Although oral non heme iron is infrequently sufficient to maintain iron stores in hemodialysis patients, recent studies suggest that heme-iron may be more useful in this regard. Heme-iron is absorbed to a greater extent than non-heme iron, and is better tolerated. Small studies have shown that when heme-iron is administered, less parenteral iron and lower doses of erythropoietin (EPO) are needed to maintain target hemoglobin. Current evidence suggests that both iron sucrose and iron gluconate are safer than iron dextran, and the latter should only be used in extraordinary circumstances. While in vitro studies have demonstrated some differences in the effects of iron sucrose and iron gluconate on cellular toxicity, the clinical importance of these has not been determined. Both compounds can be used safely for repletion and maintenance therapy, and doses of up to 300 mg of either are generally well tolerated when such higher doses are needed, as in peritoneal dialysis (PD) patients or chronic kidney disease (CKD) patients not on dialysis. CONCLUSION: A number of questions remain regarding the appropriate use, efficacy, and potential toxicity of iron therapy in dialysis patients. Further prospective research should address the myriad questions raised in this review. PMID- 14531777 TI - The safety and efficacy of an accelerated iron sucrose dosing regimen in patients with chronic kidney disease. AB - BACKGROUND: Provision of adequate iron to support erythropoiesis in patients with chronic kidney disease (CKD) is time consuming and may present adherence problems for patients in the outpatient setting. We studied an accelerated regimen of high dose intravenous iron sucrose therapy in a cohort of iron-deficient, anemic CKD patients. METHODS: Intravenous iron sucrose 500 mg was infused over three hours on two consecutive days in 107 CKD patients (glomerular filtration rate, 32.3 +/- 19.6 mL/min/1.73m2, baseline hemoglobin 10.2 +/- 1.7 g/dL). Iron indices (transferrin saturation, ferritin) were measured at baseline and at two and seven days after completion of the iron regimen. Blood pressures were monitored immediately prior to, and hourly throughout the iron sucrose infusions. RESULTS: Transferrin saturation and serum ferritin increased from 18.5 +/- 8.5% and 177 +/ 123.8 ng/mL at baseline to 40.2 +/- 22.3% and 811 +/- 294.1 ng/mL in 102 evaluated patients (P < 0.015). In 55 patients with additional measurements at 7 days post-dosing, the transferrin saturation and ferritin had fallen to 26.3 +/- 10.6% and 691 +/- 261.8 ng/mL (P < 0.015 compared to two days' post-dose). Blood pressure rose slightly, but not significantly, throughout the infusions, and altering the infusion rate was not necessary. Two patients had seven adverse events that were considered related to iron sucrose. CONCLUSION: An accelerated regimen of high-dose intravenous iron sucrose therapy in CKD patients is safe and effective in restoring iron stores, and may potentially save time and improve patient adherence. PMID- 14531778 TI - Erythropoeitin dose variation in different facilities in different countries and its relationship to drug resistance. AB - BACKGROUND: The correction of anemia using erythropoeitin (EPO) is accorded high priority in the management of patients undergoing hemodialysis (HD). Target hemoglobin (Hb) levels have been established in many countries. Following an observation that the mean facility EPO dose in a chain of facilities in the United States varied by more than two-fold, an examination of the practice of anemia correction in other settings was carried out. METHODS: We reviewed demographic and laboratory parameters in prevalent HD patients in 50 United States facilities and in a single HD facility in Vicenza, Italy. The mean EPO dose profile of the United States facilities was compared with the profiles in 10 facilities in the eastern United Kingdom (UKER) and in 20 facilities reporting to the United Kingdom Renal Registry (UKRR). Analysis of the factors that correlate with EPO resistance was carried out using the United States and Italian data. RESULTS: The average EPO doses, by facility, in the 51 United States, the 10 UKER, and the 19 UKRR facilities were 19,569, 8,416, and 7,992 international units per week (IU/wk), respectively. While examination of the UKRR revealed a similar degree of inter-facility variation (2.6-fold), much larger doses of EPO were being administered in the United States patients, particularly in the low Hb group. Multivariate analysis of the United States data suggested that factors related to inflammation, including low albumin, the use of tunneled catheters for vascular access, and low protein catabolic rate (enPCR) correlated with low Hb and relative EPO resistance. CONCLUSION: Despite similar guidelines for anemia management, significant differences in practice are observed. While there seems to be a reluctance to administer large EPO doses to individual patients in Europe, this does not seem to apply in the United States, where more EPO is given. EPO resistance seems relative rather than absolute in many patients, allowing some to respond to the higher doses. PMID- 14531779 TI - Malnutrition and inflammation in peritoneal dialysis patients. AB - BACKGROUND: Malnutrition, cardiovascular disease, and heightened inflammation are highly prevalent in dialysis patients, and major contributors to morbidity and mortality. We have investigated the inter-relationship between malnutrition and inflammation, and their impact on morbidity and mortality in peritoneal dialysis (PD) patients. METHOD: We enrolled 63 PD patients beginning in November 2000, and measured C-reactive protein (CRP) and various nutritional markers, including prealbumin. RESULTS: CRP level was elevated in 29% of the PD patients. Diabetics had higher CRP than non-diabetics (24 vs. 9.3 mg/L, P = 0.016). Patients who were hospitalized during the study had higher enrollment CRP (16 vs. 12.5 mg/L, P = 0.05) and lower enrollment albumin (3.5 vs. 3.9 g/dL, P = 0.002), blood urea nitrogen (BUN) (40 vs. 49 mg/dL, P = 0.034), and protein catabolic rate (nPCR) (0.88 vs. 1.0 g/kg/day, P = 0.02) than those who were not hospitalized. Enrollment level of CRP was inversely correlated with nutritional markers prealbumin (r = -0.5, P < 0.0001) and creatinine (r =-0.35, P < 0.01). After adjusting for age, race, gender, diabetes, and CRP level, prealbumin continued to correlate with other nutritional markers. There was a trend toward association of elevated CRP with all-cause mortality in PD patients. CONCLUSION: It is useful to incorporate prealbumin and CRP in the regular assessment of PD patients, whose survival may be improved by better management of malnutrition and inflammation. PMID- 14531780 TI - Serum albumin concentration in dialysis patients: why does it remain resistant to therapy? AB - Serum albumin, transferrin, and prealbumin levels decrease as glomerular filtration rate (GFR) declines, even prior to the start of dialysis. The levels of these serum proteins are also associated with creatinine levels and lean body mass. Lean body mass also decreases with advancing renal failure. While all of these measures are regarded as reflections of nutritional status, each are strongly associated with any of several indicators of inflammation: positive acute-phase proteins or the cytokines that regulate their synthesis rate, in both longitudinal and cross-sectional studies. Inflammation in turn is associated with comorbid conditions, cardiovascular disease, chronic infections, age, and vascular access type. Additionally, dialysis patients are subjected to oxidative stress and exposure of blood to foreign antigens in the dialysis process that also potentially contribute to inflammation. In otherwise healthy individuals reduced protein and calorie intake does not cause hypoalbuminemia since albumin fractional catabolic rate (FCR) and resting energy expenditure (REE) normally decrease in response. The simultaneous occurrence of decreased protein intake and inflammation prevent these homeostatic compensations to reduced nitrogen and energy intake from occurring, resulting in decreasing albumin, transferrin, and prealbumin levels and loss of muscle mass. Nutritional intake may also be challenged as a result of renal failure associated with anorexia, gastroparesis, and socioeconomic factors, which may all cause nutritional intake to be sufficiently marginal so that the combined effects of inflammation and decrease protein intake are expressed as decreased visceral and somatic protein stores. PMID- 14531781 TI - Transforming growth factor beta and progression of renal disease. AB - End-stage renal disease (ESRD) is more frequent in African Americans compared to whites. Many factors may be responsible, including genetic differences, increased prevalence of risk factors, and socioeconomic factors; however, to date, these proposed genetic or environmental factors have not provided a satisfactory explanation for the increased risk of ESRD in African Americans. Because renal fibrosis is a correlate of progressive renal failure and a dominant feature of ESRD, and because transforming growth factor beta 1 (TGF-beta1) can induce fibrosis and renal insufficiency, we explored the hypothesis that TGF-beta1 hyperexpression is more frequent in African Americans compared to whites. We tested our hypotheses by measuring TGF-beta1 levels in African Americans and white patients with ESRD, hypertension, and in normal patients. In hypertensive and normal patients, we also evaluated TGF-beta1 mRNA levels, and TGF-beta1 DNA polymorphisms. We demonstrated that circulating levels of TGF-beta1 are higher in African American ESRD patients, hypertensive patients, and normal control patients compared to their white counterparts. We also reported that TGF-beta1 mRNA levels are higher in hypertensives compared to normotensives. Our preliminary genetic analyses suggest that TGF-beta1 DNA polymorphisms may distinguish hypertensives from normotensives, and our laboratory is currently investigating racial differences in TGF-beta1 DNA polymorphisms. Our observations of hyperexpression of TGF-beta1 in African Americans suggest a mechanism for the increased prevalence of renal failure and hypertensive target organ damage in this population. PMID- 14531783 TI - Overview of renal bone disease: causes of treatment failure, clinical observations, the changing pattern of bone lesions, and future therapeutic approach. PMID- 14531782 TI - Obstructive nephropathy and renal fibrosis: The role of bone morphogenic protein 7 and hepatocyte growth factor. AB - BACKGROUND: The nephropathy induced by ureteral obstruction is associated with increased interstitial volume due to matrix deposition, fibroblast differentiation/proliferation, and monocyte infiltration. Recent studies indicate that transforming growth factor-beta (TGF-beta) is linked to renal fibrosis. Tumor necrosis factor (TNF-alpha) has a role in the recruitment of inflammatory cells. We found that infiltration of macrophages of the interstitium in unilateral ureteral obstruction (UUO) occurred as early as four hours after the onset of UUO. METHODS: Recent studies indicate that a renal tubular development morphogen, bone morphogenetic protein-7 (BMP-7), is effective in preventing the tubulointerstitial nephritis in the setting of obstructive nephropathy. The mechanism of action appears to be preservation of epithelial cell phenotype, inhibition of epithelial-mesenchymal transdifferentiation, and inhibition of injury-induced epithelial cell apoptosis. Hepatocyte growth factor (HGF) also inhibited tubulointerstitial fibrosis. RESULTS: In a treatment protocol in rats with ureteral ligation, BMP-7 restored renal function. The preservation of glomerular filtration rate (GFR) was accompanied by a significant decrease in cortical interstitial volume. In diabetic rats given BMP-7 proteinuria was normalized. In mice with ureteral obstruction, HGF suppressed the expression of TGF-beta and of platelet-derived growth factor. The onset of tubulointerstitial fibrosis was almost completely inhibited by HGF. CONCLUSION: Both BMP-7 and HGF attenuate the tubulointerstitial fibrosis due to ureteral obstruction. They also increase GFR and renal plasma flow. PMID- 14531784 TI - New assays for parathyroid hormone (PTH) and the relevance of PTH fragments in renal failure. AB - BACKGROUND: Immunometric assays for parathyroid hormone (PTH) are used extensively to assess bone and mineral metabolism in patients with end-stage renal disease (ESRD) who are treated with dialysis. Results generally correspond to bone histology as documented by bone biopsy, and they are useful in monitoring disease progression. Recent work has shown, however, that older, first-generation immunometric PTH assays detect not only full-length PTH(1-84), but also other amino-terminally-truncated PTH fragments (ntPTH) that may have inhibitory effects on bone cell metabolism and/or contribute to the development of adynamic renal osteodystrophy. New second-generation immunometric PTH assays, by contrast, detect PTH(1-84) exclusively. The diagnostic value of plasma PTH determinations using second-generation immunometric PTH assays and the utility of estimates of the concentration of ntPTH in plasma in patients with ESRD has been assessed only recently. METHODS: Results were reviewed from three published studies that examined the relationship between bone histology and plasma PTH levels as measured both by first- and by second-generation immunometric PTH assays in patients with ESRD. In all three studies, the concentration of ntPTH was estimated from the numerical difference between the results obtained with each assay and a ratio of PTH(1-84)/ntPTH was calculated. RESULTS: In one report, all patients with adynamic renal osteodystrophy had PTH(1-84)/ntPTH ratio values <1.0, although some patients with high-turnover skeletal lesions also had values <1.0. Estimates of the ratio of PTH(1-84)/ntPTH were found to be a better predictor of adynamic bone than PTH values measured by either assay. By contrast, two other studies failed to confirm these observations. One made use of the same second-generation immunometric PTH assay employed in the original report, whereas the other used a different assay with similar specificity for PTH(1-84). Plasma PTH levels obtained by first- and second-generation assays were highly correlated in these two independent reports. CONCLUSION: Plasma PTH levels, as determined by first-generation and second-generation immunometric assays, are highly correlated and have similar diagnostic value for the non-invasive assessment of renal osteodystrophy. The contention that ntPTH estimates and values for the PTH(1 84)/ntPTH ratio are useful in the diagnostic assessment of renal osteodystrophy has yet to be confirmed. PMID- 14531785 TI - An update on vitamin D as related to nephrology practice: 2003. AB - In this article, an up-to-date consideration of vitamin D therapeutics in nephrology is reviewed. The condition of vitamin D insufficiency is defined as the level of serum 25(OH)vitamin D at which vitamin D2 or D3 supplementation leads to a reduction of levels of parathyroid hormone (PTH). The risks of such vitamin D insufficiency in the normal population and likely risks in individuals with chronic kidney disease (CKD) stages 3 and 4 are reviewed. The potential for its safe treatment and prevention using moderate supplements of vitamin D2 or vitamin D3 are outlined. The role of altered "vitamin D nutrition" in leading to the observed greater incidence of secondary hyperparathyroidism in African Americans with ESRD compared to other racial groups is considered. The actions of active vitamin D sterols to augment intestinal absorption of both calcium and phosphorus, the effect to reduce levels of PTH, and to be a factor contributing to the rising incidence of low bone turnover (adynamic bone) are discussed. Growing evidence for contributions of elevated levels of serum calcium, serum phosphorus, and the calcium x phosphorus product as factors contributing to vascular and cardiac calcification in ESRD patients are cited. Questions are raised about whether the current practice of vitamin D usage in ESRD patients might be a contributing factor to such vascular abnormalities. The economic factors that likely affect the usage of intravenous vitamin D sterols in the United States are reviewed. It is recommended that potential adverse vascular effects of vitamin D sterols related to the increments of serum Ca and P be carefully evaluated. PMID- 14531786 TI - The impact of calcimimetics on mineral metabolism and secondary hyperparathyroidism in end-stage renal disease. AB - The impact of calcimimetics on mineral metabolism and secondary hyperparathyroidism in end-stage renal disease. Secondary hyperparathyroidism is often complicated by elevations in calcium and phosphorus either as a result of the disease per se or due to toxicity from current therapeutic options. These disturbances in mineral metabolism limit the successfulness of therapy and have been implicated as contributing to the development and progression of vascular calcification, an important and often overlooked component of cardiovascular disease in patients on dialysis. Phosphorus, active vitamin D, and calcium all play important roles in the pathogenesis of secondary hyperparathyroidism; however, serum calcium is the primary regulator of minute-to-minute parathyroid hormone secretion. Small changes in serum calcium are detected by a cell surface calcium sensing receptor that has recently been cloned. Calcimimetic agents modulate the activity of the calcium-sensing receptor and result in profound reductions in levels of circulating parathyroid hormone. Additionally, these agents result in decreases in serum calcium, phosphorus, and calcium-phosphorus product. Recently completed phase 2 clinical trials with the second-generation calcimimetic agent cinacalcet HCl confirm that this agent represents a safe and effective novel therapeutic agent which has the potential to dramatically alter the treatment and complications associated with secondary hyperparathyroidism in patients on dialysis. PMID- 14531787 TI - Plasticity of kidney cells: role in kidney remodeling and scarring. AB - The progression of renal scarring and the associated loss of function remains one of the main challenges in nephrology. Until recently, the glomerular and tubulointerstitial scarring processes were thought to involve primarily interactions between infiltrating inflammatory cells and resident renal cells culminating in loss of renal cells and their replacement by extracellular collagenous matrix (ECM). This review focuses on new aspects of renal response to injury and remodeling. Emphasis is on the plasticity of renal cells with the capacity of both glomerular and tubular cells to assume a range of phenotypes during the remodeling process. Both glomerular and tubular epithelial cells regress to primitive/embryonic mesenchymal phenotype in response to injury. This reverse embryogenesis is a key step in renal healing and scarring. In addition to the plasticity of intrinsic renal cells, it is becoming apparent that renal remodeling in health and disease involves the migration of progenitor hematopoietic stem cells into the kidneys. These cells assume various glomerular and tubular epithelial phenotype. They are also involved in the evolution of lesions toward healing or scarring. A better understanding of some of these key events in renal remodeling and their mediators may open the way to new interventions based on their manipulations and aimed at favoring renal healing and preventing scarring. PMID- 14531788 TI - Significant contribution of genomic rearrangements in SLC3A1 and SLC7A9 to the etiology of cystinuria. AB - BACKGROUND: Cystinuria is an inherited disorder of defective renal reabsorption of cystine and the dibasic amino acids. Recently, SLC3A1 and SLC7A9 have been identified as responsible genes. While point mutations in the two genes are well known to cause cystinuria, only a few studies are aimed on the identification of gross genomic alterations. Here, we report our results of a systematic screening for deletions and duplications in SLC3A1 and SLC7A9 by quantitative real-time polymerase chain reaction (PCR). METHODS: We screened a cohort of 49 cystinurics for copy number deviations in the genes SLC3A1 and SLC7A9 by quantitative real time PCR assays using fluorogenic 5' nuclease chemistry. The detected duplication in SLC3A1 was analyzed in detail by further real-time assays, reverse transcription (RT)-PCR and direct sequencing. RESULTS: In seven patients, we could identify a large duplication in SLC3A1 spanning from intron 4 to intron 9. This tandem duplication was accompanied by a small inversion of 25 bp and a 2 bp deletion in intron 9. As a formation mechanism, we presume that the inversion in intron 9 and several Alu sequences neighbored to the affected region provoke a chromatin structure that stimulates the duplication event. In addition to the SLC3A1 duplication, we observed deletions in SLC7A9 in three patients. CONCLUSION: The frequency of genomic rearrangements in our patient population illustrates the significant contribution of large genomic alterations to the mutation spectrum in cystinuria. As we could show, quantitative real-time PCR is a reliable and effective tool for the identification of unbalanced genomic rearrangements. PMID- 14531789 TI - EGF receptor tyrosine kinase inhibition attenuates the development of PKD in Han:SPRD rats. AB - BACKGROUND: Increasing evidence supports an important role for the epidermal growth factor (EGF)/transforming growth factor-alpha (TGF-alpha)/EGF receptor (EGFR) axis in promoting tubular epithelial cell proliferation and cyst formation in polycystic kidney disease (PKD). METHODS: To determine whether the inhibition of EGFR tyrosine kinase activity can attenuate the development of PKD in the Han:SPRD rat, a frequently used animal model of autosomal-dominant slowly progressive PKD (ADPKD), wild-type and cy/+ rats were treated with EKI-785 or EKB 569 or with vehicle alone. Western analysis, immunoprecipitation, and immunohistochemistry were used to ascertain the expression, activation, and localization of EGFR. RESULTS: Overexpression, activation and apical mislocalization of EGFR were observed in the cy/+ rats. The intraperitoneal administration of EKI-785 reversed the activation of the EGFR to the level observed in wild-type animals. The intraperitoneal administration of EKI-785 (90 mg/kg body weight every third day) or of EKB-569 (20 mg/kg body weight every third day) to cy/+ rats resulted in lower kidney weights, serum concentrations of blood urea nitrogen (BUN), cyst volumes, and fibrosis scores. The administration of EKB-569 by gavage was less effective probably because of lower bioavailability. CONCLUSION: These results support a significant role for the EGF/TGF-alpha/EGFR axis in the development of PKD in the Han:SPRD rat and the therapeutic potential of EGFR tyrosine kinase inhibition in ADPKD. PMID- 14531790 TI - Mutations of the Uromodulin gene in MCKD type 2 patients cluster in exon 4, which encodes three EGF-like domains. AB - BACKGROUND: Autosomal-dominant medullary cystic kidney disease type 2 (MCKD2) is a tubulointerstitial nephropathy that causes renal salt wasting, hyperuricemia, gout, and end-stage renal failure in the fifth decade of life. The chromosomal locus for MCKD2 was localized on chromosome 16p12. Within this chromosomal region, Uromodulin (UMOD) was located as a candidate gene. UMOD encodes the Tamm Horsfall protein. By sequence analysis, one group formerly excluded UMOD as the disease-causing gene. In contrast, recently, another group described mutations in the UMOD gene as responsible for MCKD2 and familial juvenile hyperuricemic nephropathy (FJHN). METHODS: Haplotype analysis for linkage to MCKD2 was performed in 25 MCKD families. In the kindreds showing linkage to the MCKD2 locus on chromosome 16p12, mutational analysis of the UMOD gene was performed by exon polymerase chain reaction (PCR) and direct sequencing. RESULTS: In 19 families, haplotype analysis was compatible with linkage to the MCKD2 locus. All these kindreds were examined for mutations in the UMOD gene. In three different families, three novel heterozygous mutations in the UMOD gene were found and segregated with the phenotype in affected individuals. Mutations were found only in exon 4. CONCLUSION: We confirm the UMOD gene as the disease-causing gene for MCKD2. All three novel mutations were found in the fourth exon of UMOD, in which all mutations except one (this is located in the neighboring exon 5) published so far are located. These data point to a specific role of exon 4 encoded sequence of UMOD in the generation of the MCKD2 renal phenotype. PMID- 14531791 TI - A catalogue of gene expression in the developing kidney. AB - BACKGROUND: Although many genes with important function in kidney morphogenesis have been described, it is clear that many more remain to be discovered. Microarrays allow a more global analysis of the genetic basis of kidney organogenesis. METHODS: In this study, Affymetrix U74Av2 microarrays, with over 12,000 genes represented, were used in conjunction with robust target microamplification techniques to define the gene expression profiles of the developing mouse kidney. RESULTS: Microdissected murine ureteric bud and metanephric mesenchyme as well as total kidneys at embryonic day E11.5, E12.5, E13.5, E16.5, and adult were examined. This work identified, for example, 3847 genes expressed in the E12.5 kidney. Stringent comparison of the E12.5 versus adult recognized 428 genes with significantly elevated expression in the embryonic kidney. These genes fell into several functional categories, including transcription factor, growth factor, signal transduction, cell cycle, and others. In contrast, surprisingly few differences were found in the gene expression profiles of the ureteric bud and metanephric mesenchyme, with many of the differences clearly associated with the more epithelial character of the bud. In situ hybridizations were used to confirm and extend microarray-predicted expression patterns in the developing kidney. For three genes, Cdrap, Tgfbi, and Col15a1, we observed strikingly similar expression in the developing kidneys and lungs, which both undergo branching morphogenesis. CONCLUSION: The results provide a gene discovery function, identifying large numbers of genes not previously associated with kidney development. This study extends developing kidney microarray analysis to the powerful genetic system of the mouse and establishes a baseline for future examination of the many available mutants. This work creates a catalogue of the gene expression states of the developing mouse kidney and its microdissected subcomponents. PMID- 14531792 TI - The tyrosine kinase inhibitor tyrphostin AG126 reduces renal ischemia/reperfusion injury in the rat. AB - BACKGROUND: We investigate the effects of tyrphostin AG126, an inhibitor of tyrosine kinase activity, on the renal dysfunction and injury caused by ischemia/reperfusion (I/R) of the kidney. METHODS: Tyrphostin AG126 (5 mg/kg intraperitoneally) was administered to male Wistar rats 30 minutes prior to bilateral renal ischemia for 45 minutes followed by reperfusion for up to 48 hours. Biochemical markers of renal dysfunction and injury were measured and renal sections assessed for renal injury. Expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) and formation of nitrotyrosine and poly (ADP) ribose (PAR) were assessed using immunohistochemistry. Rat proximal tubular cells (PTCs) were incubated with interferon-gamma (100 IU/mL), bacterial lipopolysaccharide (10 microg/mL), and with increasing concentrations of tyrphostin AG126 (0.0001-1 mmol/L) for 24 hours. Nitric oxide production was measured in both plasma from rats subjected to I/R and in incubation medium from PTCs. RESULTS: After 6 hours of reperfusion, tyrphostin AG126 significantly reduced the increase in serum and urinary indicators of renal dysfunction and injury caused by I/R and reduced histologic evidence of renal injury. Tyrphostin AG126 also improved renal function (after 24 and 48 hours of reperfusion) and reduced the histologic signs of renal injury (after 48 hours of reperfusion). Tyrphostin AG126 reduced the expression of iNOS and nitric oxide levels in both rat plasma and in PTC cultures, as well as expression of COX-2. Tyrphostin AG126 also reduced nitrotyrosine and PAR formation, suggesting reduction of nitrosative stress and poly (ADP-ribose) polymerase (PARP) activation, respectively. CONCLUSION: Taken together, these results show that tyrphostin AG126 significantly reduces the renal dysfunction and injury caused by I/R of the kidney. We propose that inhibition of tyrosine kinase activity may be useful against renal I/R injury. PMID- 14531793 TI - Ethyl pyruvate decreases sepsis-induced acute renal failure and multiple organ damage in aged mice. AB - BACKGROUND: Sepsis is a common cause of acute renal failure (ARF). The incidence of sepsis increases dramatically after 50 years of age; however, most ARF studies are performed in young mice. METHODS: We performed two common sepsis models, lipopolysaccharide (LPS) administration and cecal ligation puncture (CLP) in aged mice. We developed a fully treated CLP model in aged mice by treating mice with fluid resuscitation and antibiotics. RESULTS: LPS induced renal injury in aged but not young mice. However, volume resuscitation starting within 6 hours decreased renal injury. We then used this fluid resuscitation scheme, along with antibiotics, to develop a fully treated CLP model in aged mice. Mice subjected to CLP developed functional and histologic ARF and multiple organ damage. Treatment with ethyl pyruvate, even when started 12 hours after surgery, decreased serum creatinine, tubular damage, and multiple organ injury at 24 hours. Ethyl pyruvate decreased plasma tumor necrosis factor-alpha (TNF-alpha), and kidney mRNA for TNF alpha, tissue factor, and plasminogen activator inhibitor-1 (PAI-1), and increased mRNA for urokinase-like plasminogen activator. CONCLUSION: CLP in aged mice causes functional and histologic changes consistent with human ARF. A single dose of ethyl pyruvate inhibits renal and multiple organ damage, and is still effective when given 12 hours after surgery. PMID- 14531794 TI - Vitamin D3 up-regulated protein-1 regulates collagen expression in mesangial cells. AB - BACKGROUND: Hyperglycemia is a known risk factor in the pathogenesis of nephropathy, and collagen accumulation due to an increase reactive oxygen species (ROS) has been suspected to be one of the reasons for high glucose-mediated diseases. However, molecular mechanisms that connect glucose stimulation, oxidative stress, and collagen induction are unknown. METHODS: We examined global changes in gene expression patterns following high glucose stimulation by using DNA microarray technology in cultured human mesangial cells. The expression of vitamin D3 up-regulated protein-1 (VDUP-1), our candidate for the molecular mediator, was evaluated in the human mesangial cells, mouse mesangial cell line, and kidneys of diabetic mice by quantitative reverse transcription-polymerase chain reaction (RT-PCR). Truncated VDUP-1 proteins were used to test the effects of VDUP-1 on the biosynthesis of collagen in mesangial cells. RESULTS: Expression of VDUP-1, which was reported as an inhibitor of thioredoxin, was induced rapidly and constantly after exposure to high concentrations of glucose upon analysis with DNA microarray. Overexpression of VDUP-1 gene in cultured mesangial cells resulted in type IV collagen alpha1 chain (COL4A1) mRNA induction and accumulation of type IV collagen protein. However, induction of COL4A1 expression was abolished with a deletion mutant of VDUP-1, which lost thioredoxin interacting domain. Also, streptozotocin-induced diabetic mice were shown to overexpress VDUP-1 as well as COL4A1. CONCLUSION: VDUP-1 mediates collagen accumulation in mesangial cells and could be the molecular mediator/marker for fibrosis in diabetic nephropathy caused by chronic hyperglycemia such as diabetes. PMID- 14531795 TI - Role of plasma renin activity in the management of primary vesicoureteric reflux: a preliminary report. AB - BACKGROUND: Nonoperative management is the choice of therapy in most patients with primary vesicoureteric reflux (VUR). Such patients are regularly monitored for breakthrough urinary tract infection, deterioration of renal function, and progression/ appearance of new scars as the indicators for switching over to surgical intervention. In this descriptive study, we report our additional observations on serial measurement of plasma renin activity (PRA) in a group of such children followed prospectively. METHODS: Twenty-six children (16 males and 10 females) with various grades (grades I to V) of primary VUR were enrolled. Besides the conventional investigations, we also monitored PRA. Ureteric reimplantation was performed based on established international criteria. PRA levels were correlated with the need for surgical intervention retrospectively. RESULTS: Mean age at presentation was 39.3 months (range, 12 to 120 months). Nineteen children (73%) were subjected to ureteric reimplantation. The mean PRA in this group (N = 19) was 6.97 ng/mL/hour versus 3.28 ng/mL/hour in patients who were continued on nonoperative management (N = 7). Postoperatively, the PRA reduced and stabilized in all 18 patients at a mean value of 5.4 ng/mL/hour. CONCLUSION: The currently accepted end points of medical management are inconsistent. On the other hand, 94.7% of children in the surgical group had shown an activation and a progressive increase in PRA. Postoperatively, the PRA was reduced and stabilized in all these patients. Our preliminary observations suggest that high PRA is a more sensitive indicator for surgical intervention as compared to the existing criteria. PMID- 14531796 TI - Endothelial progenitor cell proliferation and differentiation is regulated by erythropoietin. AB - BACKGROUND: Circulating bone marrow-derived endothelial progenitor cells (EPCs) promote vascular reparative processes. In humans, their number correlate with endothelial function and cardiovascular risk. We tested the hypothesis that darbepoetin alfa [i.e., a recombinant analogue of the cytokine erythropoietin (EPO)] stimulates proliferation and differentiation of EPCs. METHODS: We assessed CD34+ circulating stem cells (cSCs) in whole blood using flow cytometry and, in addition, proliferation/differentiation of EPCs in an in-vitro assay during 6 weeks of a standard darbepoetin therapy in eight patients with renal anemia. RESULTS: Darbepoetin treatment caused a significant increase in the number of CD34+ cSCs (week 2, 193%+/- 46%; and week 6, 298%+/- 90%; P < 0.05 vs. baseline). In addition, darbepoetin markedly increased the number of functionally active EPCs (week 2, 256%+/- 48%; and week 6, 299%+/- 59%; both P < 0.01 vs. baseline). The effect of darbepoetin on functional activity of EPCs assessed in a tube formation assay was dose dependent. Administration of darbepoietin caused activation of protein kinase B (Akt) in cultured EPCs. CONCLUSION: A standard treatment with darbepoetin markedly enhances EPC proliferation and differentiation in renal patients. The use of recombinant EPO analogues may be a novel and safe therapeutic approach in patients with vascular pathology. PMID- 14531798 TI - Internalization of antibodies by endothelial cells via fibronectin implicating a novel mechanism in lupus nephritis. AB - BACKGROUND: One of the crucial events in lupus nephritis is the glomerular deposition of immunoglobulins (Igs), of which pathogenic properties have been proposed mostly to be either type IIor type III allergic reactions. Some of IgG3 producing hybridoma clones established from an MRL/MpTn-gld/gld (MRL/gld) lupus mouse generate wire loop-like lesions in glomeruli resembling lupus nephritis when injected into SCID mice. These clones are useful for analyzing the mechanisms of glomerular deposition of antibodies in lupus nephritis at the monoclonal level. METHODS: Glomerular lesions of SCID mice injected with the hybridoma clones, 17H8a or 1G3 as control were analyzed by light and electron microscopy. Interaction of the antibodies with human glomerular endothelial cells (HGECs) and human umbilical vein endothelial cells (HUVECs) in vitro was studied by fluorescence microscopy, electron microscopy, and flow cytometry. RESULTS: Both antibodies did not show any antigen specificity for mouse glomeruli. The glomerular lesions generated by 17H8a, but not by 1G3, contained electron-dense deposits not only in subendothelial regions but also in the cytoplasm of endothelial cells, suggesting internalization of the 17H8a antibodies by endothelial cells. In cell culture studies, internalization of only 17H8a antibodies by HGECs and HUVECs was observed, but the antibodies did not have antigen specificity for both types of endothelial cells. The internalization by HUVECs was mediated by actin polymerization, and it was inhibited by RGDS (Arg Gly-Asp-Ser) tetrapeptide, antihuman fibronectin and antihuman integrin beta1 monoclonal antibodies. CONCLUSION: The interaction between particular antibodies and endothelial cell surface integrins via fibronectin may be involved in their subsequent internalization by endothelial cells leading to antibody deposition in glomeruli. This may be one of the mechanisms of glomerular injury in lupus nephritis. PMID- 14531797 TI - Sevelamer hydrochloride attenuates kidney and cardiovascular calcifications in long-term experimental uremia. AB - BACKGROUND: In chronic renal failure (CRF), hyperphosphatemia and an elevated calcium-phosphate product are associated with vascular calcification and increased cardiovascular morbidity and mortality. Previous data have demonstrated that 3-month treatment of uremic rats with sevelamer was associated with less nephrocalcinosis compared to calcium carbonate (CaCO3), despite similar control of serum phosphorus, calcium-phosphorus product (Ca x P product), and secondary hyperparathyroidism. There was no evidence of aortic calcification after 3 months of uremia (J Am Soc Nephrol 13:2299-2308, 2002). The present studies explore the influence of sevelamer and CaCO3 on cardiovascular and kidney calcifications in long-term experimental uremia over 6 months. METHODS: Normal and 5/6 nephrectomized rats (U) were fed a high phosphorus (HP) diet for 6 months. Two phosphate binders, CaCO3 and sevelamer, were administered and their influence on hyperphosphatemia, secondary hyperparathyroidism, kidney/myocardial/aortic calcification, and renal function was compared. RESULTS: All uremic rats began the study with the same degree of renal failure. Sevelamer was as effective as CaCO3 in reducing serum phosphorus, Ca x P product, and attenuating secondary hyperparathyroidism. Despite similar serum cholesterol levels, rats in the U-HP + sevelamer group had markedly lower calcium deposition in the myocardium and aorta (myocardium, 72 +/- 4 microg/g wet tissue; aorta, 736 +/- 156 microg/g wet tissue) compared to rats in either the U-HP + CaCO3 group (myocardium, 179 +/- 48, P < 0.05; aorta, 1308 +/- 343, P < 0.05) or the U-HP group (myocardium, 98 +/ 10, NS; aorta, 2150 +/- 447, P < 0.05). Dual immunohistochemical analysis for calcium and endothelial cell markers demonstrated that myocardial calcium deposition was intravascular within capillaries. Furthermore, calcium deposition in the kidney of uremic rats treated with sevelamer (582 +/- 111 microg/g wet tissue) was lower than that found in uremic rats treated with CaCO3 (1196 +/- 180 microg/g wet tissue). Sevelamer-treated rats had less deterioration in renal function with an associated lower serum creatinine, higher creatinine clearance, and less proteinuria. There was no difference in overall mortality between the three experimental groups. CONCLUSION: In long-term experimental CRF, in addition to controlling serum phosphorus and secondary hyperparathyroidism as efficiently as CaCO3, treatment with the phosphate-binder sevelamer attenuates vascular and kidney calcification. PMID- 14531799 TI - VHL down-regulation and differential localization as mechanisms in tumorigenesis. AB - BACKGROUND: The von Hippel-Lindau (VHL) gene has been widely analyzed in many tumors. Early studies in animal tumors suggest that changes in VHL protein level and localization may be also important in tumorigenesis. In this study, we determined the role of VHL protein in human renal cell carcinomas. METHODS: Seventy-five human renal cell carcinomas, predominantly of clear cell type (60 of 75), were examined for VHL protein by immunohistochemistry. The level and pattern of protein expression were then compared to VHL mutations and tumor characteristics. RESULTS: An apparent decline of VHL level (positive in <50% of tumor cells) was observed in 49 (65%) tumors, a change more frequent than VHL mutations (28 of 75) (37%). In tumors, VHL was localized to the cytoplasm and/or the cell membrane. The occurrence of a predominantly membranous signal was significantly associated with missense mutations (9 of 14 tumors with missense mutations versus 14 of 61 tumors with no or nonmissense mutations, P = 0.0025) and tumor stage (23 of 60 tumors with stage TI versus 0 of 15 tumors with TII and TIII, P = 0.0034). CONCLUSION: This study provides the first evidence of the role of VHL protein level and intracellular localization in tumorigenesis in humans. PMID- 14531800 TI - Differential effects of Sendai virus infection on mediator synthesis by mesangial cells from two mouse strains. AB - BACKGROUND: Recently, we observed that the severity of glomerulonephritis in an experimental model of immunoglobulin A nephropathy (IgAN) induced by Sendai virus differs between C57BL/6 and BALB/c mouse strains. The determinants of differing renal insufficiency are not understood. In the present study, we examine the capacity for mesangial cells to support Sendai viral replication and assess the direct effects of Sendai virus on the production of selected cytokines, chemokines, and eicosanoids by mesangial cells, comparing C57BL/6 to BALB/c mouse strains. METHODS: Sendai virus replication was measured by viral plaque assay using LLCMK2 cells. Production of cytokines [interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha)], chemokines (JE and KC), and eicosanoids [prostaglandin E2 (PGE2) and thromboxane B2 (TxB2)] in culture medium was evaluated by sandwich enzyme-linked immunosorbent assay (ELISA) or competitive enzyme immunoassay (EIA) after 48 hours' incubation with infectious or inactivated Sendai virus. RESULTS: Sendai virus replicates equally well in mesangial cells from both strains, and infection evokes increased IL-6, JE, KC, and PGE2 production in relation to viral dose. BALB/c mesangial cells produce significantly more IL-6 and JE than those from C57BL/6, and the dose response for KC is steeper in BALB/c mesangial cells than those from C57BL/6. Synthesis of PGE2 in BALB/c mesangial cells is higher than that of C57BL/6 mesangial cells, both under basal conditions and in response to infectious Sendai virus, again in a dose-dependent manner. There is no TNF-alpha or thromboxane response to viral stimulation. CONCLUSION: We conclude that different mesangial cell responses to this common mucosal viral pathogen might influence the severity of IgAN in our model system. PMID- 14531801 TI - Regulation by CD25+ lymphocytes of autoantigen-specific T-cell responses in Goodpasture's (anti-GBM) disease. AB - BACKGROUND: Goodpasture's, or anti-glomerular basement membrane (GBM), disease is unusual among autoimmune diseases in that it rarely follows a relapsing-remitting course. Moreover, untreated, autoantibodies disappear spontaneously after 1 to 3 years and, following treatment, autoreactive T cells diminish in frequency. This suggests that operational tolerance toward the autoantigen is reestablished. However, the mechanisms underlying this have remained unclear. Recent data have suggested that a population of regulatory T lymphocytes can suppress both autoimmune and alloimmune responses in animal models and are present in normal individuals. However, to date, they have not been demonstrated to play a role in human renal autoimmune disease. METHODS: We studied the role of regulatory CD25+ cells in suppressing T-cell responses to the Goodpasture autoantigen in nine patients with Goodpasture's disease. RESULTS: At the time of acute presentation, there was no evidence of a regulatory cell population. However, from 3 months onward a population emerged, capable of suppressing the response to the Goodpasture autoantigen. Following depletion of CD25+ cells, the frequencies of autoreactive-, GBM-, or collagen alpha 3(IV)NC1-specific T cells were significantly increased (P = 0.031 by paired t test), with five of seven (71%) convalescent patients and no acute patients demonstrating regulation. CONCLUSION: These data demonstrate that, in Goodpasture's disease, regulatory CD25+ T cells play a role in inhibiting the autoimmune response. Their emergence and persistence may underlie the "single hit" nature of this condition. Understanding the conditions required for the development and propagation of these cells would allow development of novel therapeutic strategies for inducing hyporesponsiveness in autoimmune disease. PMID- 14531802 TI - Critical role for Nef in HIV-1-induced podocyte dedifferentiation. AB - The notable glomerular feature of human immunodeficiency virus (HIV)-associated nephropathy (HIVAN) is the collapse of the capillary tuft with marked glomerular epithelial cell hyperplasia. These data suggest a loss of normal podocyte function, which is associated with a loss of the podocyte differentiation markers, Wilm's tumor (WT-1), synaptopodin, podocalyxin, and common acute lymphoblastic leukemia antigen (CALLA). We have previously shown that HIV-1 expression can induce these changes in HIV-1 transgenic mice. To identify which HIV-1 gene product(s) are responsible for the phenotypic changes in podocytes, we created multiple mutated HIV-1 constructs and then pseudotyped them with vesticular stomatitis virus glycoprotein (VSVG) envelope to enhance the tropism of these mutant viruses. In addition to gag/pol, the mutant viruses lacked one of the following, env, nef, rev, vif, vpr, or vpu. In addition, we generated single gene expressing pseudotyped viruses to complement the scanning mutation approach of our viral parental construct. Murine podocytes were then infected with one of the viral constructs either lacking or expressing the various HIV-1 genes. We found that HIV-1 nef was necessary and sufficient for proliferation of podocytes and down-regulation of synaptopodin and CALLA. These data suggest that Nef induces many of the changes we observe in HIV transgenic model and, as a result, this now defines the pathway for exploration of host responses to HIV-1 infection. PMID- 14531803 TI - Differential behavior of mesangial cells derived from 12/15-lipoxygenase knockout mice relative to control mice. AB - BACKGROUND: The 12/15-lipoxygenase (12/15-LO) enzyme has been implicated in the pathogenesis of diabetic nephropathy since lipoxygenase products induce cellular hypertrophy and extracellular matrix deposition in mesangial cells. In this study, in order to determine the potential in vivo functional role of 12/15-LO in kidney disease, we compared mouse mesangial cells (MMCs) derived from 12/15-LO knockout mice with those from genetic control wild-type mice. METHODS: MMCs were isolated from wild-type and 12/15-LO knockout mice. Cellular growth, activation of mitogen-activated protein kinases (MAPKs), transcription factors, superoxide levels, and fibronectin expression were compared in the two cell types. RESULTS: Levels of the 12/15-LO product and protein were lower in MMC from 12/15-LO knockout relative to wild-type. MMCs from 12/15-LO knockout mice grew slower than wild-type cells, and also showed lower rates of tritiated thymidine and leucine incorporation (21% and 15% of wild-type, respectively, P < 0.001). Levels of superoxide and the matrix protein fibronectin were also lower in 12/15-LO knockout mice cells. Serum and angiotensin II (Ang II)-stimulated activities of p38 or ERK1/2 MAPKs, and cyclic adenosine monophosphate (cAMP)-responsive element binding protein (CREB) transcription factor were lower in 12/15-LO knockout relative to wild-type cells. In addition, DNA binding and transcriptional activities of activated protein-1 (AP-1) and CREB were lower in 12/15-LO knockout cells. Furthermore, stable 12/15-LO overexpression in MMC led to reciprocal increase in p38 MAPK activation and fibronectin expression. CONCLUSION: The differential activation of oxidant stress, specific signaling pathways, transcription factors, and growth and matrix genes may lead to reduced growth and growth factor responses in 12/15-LO knockout versus wild-type MMCs. These results provide ex vivo functional evidence for the first time that 12/15-LO activation plays a key role in mesangial cell responses associated with renal diseases such as diabetic nephropathy. PMID- 14531804 TI - Activation of the TGF-beta/Smad signaling pathway in focal segmental glomerulosclerosis. AB - BACKGROUND: Although the pathogenetic relevance of transforming growth factor beta (TGF-beta) to glomerulosclerosis is well established, it is not known whether a signal transduction cascade of TGF-beta is involved in the development of focal segmental glomerulosclerosis (FSGS), nor is it clear how TGF-beta 1 is activated during the course of FSGS formation. METHODS: We examined the expression patterns of TGF-beta 1, thrombospondin-1 (TSP-1), TGF-beta type II receptor (TGF-beta IIR), phosphorylated Smad2/Smad3, and podocyte-specific epitopes [Wilms' tumor protein-1 (WT-1) and glomerular epithelial protein-1 (GLEPP-1)] in 15 renal biopsy specimens with idiopathic FSGS and six renal biopsies with no detectable abnormalities by means of immunohistochemistry. The mRNA expression patterns of TGF-beta 1, TGF-beta IIR, and TSP-1 were further evaluated by in situ hybridization in seven biopsies. RESULTS: In the controls, immunostaining for TGF-beta 1, TSP-1, TGF-beta IIR, and phosphorylated Smad2/Smad3 was almost negligible, but an apparent signal for TGF-beta 1, TSP-1, and TGF-beta IIR mRNAs was observed in the visceral glomerular epithelial cells (GEC). In the cases of FSGS, the expression levels of TGF-beta 1, TSP-1, and TGF betaIIR proteins and mRNAs and phosphorylated Smad2/Smad3 were significantly increased, particularly in the GEC of the sclerotic segments, wherein WT-1 and GLEPP-1 were not detected. CONCLUSION: These results suggest that damage to podocyes may stimulate TGF-beta 1, TSP-1, and TGF-beta IIR expression in GEC, thereby activating the Smad signaling pathway and, in so doing, leading to overproduction of the extracellular matrix (ECM). Thus, a signal transduction cascade of the TGF-beta/Smad signaling pathway, which is activated in the GEC, appears to be involved in the development of FSGS. PMID- 14531805 TI - Selective depletion of fibroblasts preserves morphology and the functional integrity of peritoneum in transgenic mice with peritoneal fibrosing syndrome. AB - BACKGROUND: A peritoneal fibrosing syndrome (PFS) can progressively reduce peritoneal ultrafiltration during chronic peritoneal dialysis in patients with renal failure. The pathogenesis of PFS is unclear and the role of peritoneal fibroblasts has not been evaluated experimentally. METHODS: We followed the fate of fibroblasts producing PFS in a mouse model using fibroblast-specific protein 1 (FSP1) as a marker. PFS was induced by daily peritoneal infusions of chlorhexidine gluconate (CHG) saline into transgenic mice expressing the thymidine kinase (Delta tk) gene under the control of the FSP1 promoter (FSP1.Delta tk mice). To demonstrate the role of fibroblasts in PFS, we treated these FSP1.Delta tk mice with a nucleoside analogue to induce DNA chain termination and fibroblast death. RESULTS: Mice receiving peritoneal infusions of CHG saline every other day for 2 weeks developed increasing numbers of FSP1+ fibroblasts in the subserosal layers of the visceral peritoneum. Mac-3+ monocytes (macrophages) subsequently accumulated over the next 2 weeks in association with increased deposition of type I collagen and increased endothelial vascularity (CD31+) in these subserosal tissues. Since these peritoneal fibroblasts expressed monocyte chemoattractant protein-1 (MCP-1), heat shock protein 47 (HSP47), and vascular endothelial growth factor (VEGF), we suspect they were partially responsible for macrophage recruitment, matrix production, and the neoangiogenesis in the subserosal tissue. Treatment of PFS in FSP1.Delta tk transgenic mice with a nucleoside analogue selectively reduced the numbers of peritoneal fibroblasts and attenuated the attendant changes in peritoneal histology. Rescuing the peritoneal membrane from chronic thickening and neoangiogenesis by reducing the number of fibroblasts also preserved ultrafiltration. CONCLUSION: Peritoneal fibroblasts play a pivotal role in PFS, and their deletion using a fibroblasts-specific transgene was effective in preventing peritoneal fibrogenesis. PMID- 14531806 TI - PDZK1: I. a major scaffolder in brush borders of proximal tubular cells. AB - BACKGROUND: In proximal tubular cells, PDZK1 (NaPi-Cap1) has been implicated in apical expression of the Na+-dependent phosphate cotransporter (NaPi-IIa) via interaction with its C-terminus. PDZK1 represents a multidomain protein consisting of four PDZ domains and thus is believed to have a broader specificity besides NaPi-IIa. METHODS: We subjected single PDZ domains derived from PDZK1 either to yeast two-hybrid screens or yeast trap assays. Different pull-down assays and blot overlays were applied to corroborate the PDZK1-mediated interactions in vitro. Co-localization of interacting proteins with PDZK1 in proximal tubular cells was assessed by immunohistochemistry. RESULTS: In the yeast screens, the most abundant candidate protein to interact with PDZK1 was the membrane-associated protein of 17 kD (MAP17). Besides MAP17, C-terminal parts of following transporters were also identified: NaPi-IIa, solute carrier SLC17A1 (NaPi-I), Na+/H+ exchanger (NHE-3), organic cation transporter (OCTN1), chloride formate exchanger (CFEX), and urate-anion exchanger (URAT1). In addition, other regulatory factors were found among the clones, such as a protein kinase A (PKA) anchoring protein (D-AKAP2) and N+/H+ exchanger regulator factor (NHERF-1). All interactions of itemized proteins with PDZK1 were affirmed by in vitro techniques. Apart from PDZK1, strong in vitro interactions of NHERF-1 were also observed with the solute transporters (excluding MAP17) and D-AKAP2. All identified proteins were immunolocalized in proximal tubular cells, wherein all membrane proteins co-localized with PDZK1 in brush borders. CONCLUSION: We hypothesize that PDZK1 and NHERF-1 establish an extended network beneath the apical membrane to which membrane proteins and regulatory components are anchored. PMID- 14531807 TI - PDZK1: II. an anchoring site for the PKA-binding protein D-AKAP2 in renal proximal tubular cells. AB - BACKGROUND: PDZK1, a multiple PDZ protein, was recently found to interact with the type IIa Na/Pi cotransporter (NaPi-IIa) in renal proximal tubular cells. In a preceding study, yeast two-hybrid screens using single PDZ domains of PDZK1 as baits were performed. Among the identified proteins, a C-terminal fragment of the dual-specific A-kinase anchoring protein 2 (D-AKAP2) was obtained by screening PDZ domain 4. METHODS: After its molecular cloning by means of RACE, the renal expression of D-AKAP2 was analyzed by real-time polymerase chain reaction (PCR) and immunohistochemistry. Protein interactions were characterized by overlays, pull-downs, and immunoprecipitations from transfected opossum kidney (OK) cells. RESULTS: Based on 5'-RACE and PDZK1 overlays of mouse kidney cortex separated by two-dimensional electrophoresis, it was suggested that the renal isoform of D AKAP2 in mouse comprises 372 amino acids and exists as a protein of >40 kD. Immunohistochemistry and real-time PCR localized D-AKAP2 only to the subapical pole of proximal tubular cells in mouse kidney. In pull-down experiments, D-AKAP2 tightly bound PDZK1 as well as N+/H+ exchanger regulator factor (NHERF-1), but the latter with an apparent fourfold lower affinity. Similarly, His-tagged D AKAP2 specifically retained PDZK1 from mouse kidney cortex homogenate. In addition, myc-tagged D-AKAP2 and HA-tagged PDZK1 co-immunoprecipitated from transfected OK cells. CONCLUSION: We conclude that D-AKAP2 anchors protein kinase A (PKA) to PDZK1 and to a lesser extent to NHERF-1. Since PDZK1 and NHERF-1 both sequester NaPi-IIa to the apical membrane, D-AKAP2 may play an important role in the parathyroid hormone (PTH)-mediated regulation of NaPi-IIa by compartmentalization of PKA. PMID- 14531808 TI - Iron handling and gene expression of the divalent metal transporter, DMT1, in the kidney of the anemic Belgrade (b) rat. AB - BACKGROUND: We have previously shown that the rat kidney reabsorbs metabolically significant amounts of iron and that it expresses the divalent metal transporter 1, DMT1. The Belgrade (b) rat carries a mutation in DMT1 gene, which causes hypochromic, microcytic anemia due to impaired intestinal iron absorption and transport of iron out of the transferrin cycle endosome. In the duodenum of b/b rats, expression of DMT1 mRNA and protein is increased, suggesting a feedback regulation by iron stores. The aim of this study was to investigate iron handling and DMT1 expression in the kidneys of Belgrade rats. METHODS: Animals were maintained for 3 weeks on a synthetic diet containing 185 mg/kg iron (FeSO4), after which functional and molecular parameters were analyzed in male heterozygous (+/b) and homozygous (b/b) rats (N = 4 to 6 for each group). RESULTS: Serum iron concentration was significantly higher in b/b compared to +/b rats while urinary iron excretion rates were unchanged in b/b compared to +/b rats. Northern analysis using a rat DMT1 probe showed comparable mRNA levels between +/b and b/b animals. Western analysis and immunofluorescence microscopy performed using a polyclonal antibody against rat DMT1 showed that DMT1-specific immunoreactivity was almost absent in the kidneys of b/b rats compared to that seen in +/b animals. CONCLUSION: Our results indicate that the G185R mutation of DMT1 causes protein instability in the kidneys of b/b rats. Given that +/b and b/b rats excrete comparable amounts of iron, the lack of DMT1 protein is compensated by an alternative, yet to be identified, mechanism. PMID- 14531809 TI - Renal neuronal nitric oxide synthase protein expression as a marker of renal injury. AB - BACKGROUND: Animal studies suggest that nitric oxide deficiency occurs in the remnant after 5/6 removal of renal mass. The present studies investigated the time course in relation to progression of renal disease, as well as the impact on individual renal nitric oxide synthase (NOS) isoforms. METHODS: Rats were studied from 2 to 11 weeks after 5/6 ablation/infarction (A/I) of renal mass, with acceleration of progression by high protein and salt intake, in some groups. Measurements were made before sacrifice of 24-hour protein and creatinine excretion, blood was taken for creatinine and blood urea nitrogen (BUN) determination and the kidneys were investigated histologically for structural damage, abundance of endothelial NOS (eNOS) and neuronal NOS (nNOS), and in some groups for in vitro NOS activity. RESULTS: A time-dependent fall in glomerular filtration rate (GFR) and rise in proteinuria and glomerular sclerosis developed after 5/6 A/I. The nNOS abundance in cortex and medulla was decreased relative to shams, in all but the mildest injury and there was a strong, steep correlation between level of glomerular sclerosis and the degree of reduction in renal nNOS. Where measured, cortical NOS activity correlated with the nNOS abundance. In contrast, the eNOS abundance was either increased or unchanged in rats post A/I. CONCLUSION: Renal nNOS abundance was reduced in the 5/6 A/I model of renal disease when plasma creatinine> approximately 1 mg/dL and when> approximately 20% of remaining glomeruli were sclerosed. PMID- 14531810 TI - Role of excess volume in the pathophysiology of hypertension in chronic kidney disease. AB - BACKGROUND: The pathophysiology of hypertension in patients with chronic kidney disease (CKD) is largely attributed to positive sodium balance. It is unclear how loop diuretics affect fluid volume compartments, especially with respect to their antihypertensive effect. METHODS: Subjects with CKD were administered a single therapeutically equivalent dose of an oral loop diuretic (furosemide or torsemide in randomized crossover design). We measured acute volume changes over 12 hours using biophysical and hormonal biomarkers and then 24-hour ambulatory blood pressure after daily diuretic therapy for 3 weeks. RESULTS: Single-dose administration of loop diuretic decreased extracellular water (ECW) by 1.7 L [95% confidence interval (95% CI) 1.2, 2.2, P < 0.001], total body water (TBW) by 1.2 L (95% CI 0.5, 1.9, P < 0.001), and increased natural log (ln) plasma renin activity (PRA) from -1.2 +/- 1.3 ng/mL/hour to -0.5 +/- 1.5 ng/mL/hour (P < 0.001). Daily loop diuretic administration resulted in reduced ECW from 24.2 +/- 6.4 L to 22.3 +/- 5.2 L (P = 0.02) and TBW from 54.3 +/- 12.7 L to 51.6 +/- 11.9 L (P < 0.001) in 1 week. After 3 weeks of diuretic therapy, whereas ECW reduction persisted at 22.8 +/- 5.1 L (P = 0.05), TBW trended toward baseline level at 52.7 +/- 11.8 L. A concomitant increase in ln PRA from -1.0 +/- 1.3 ng/mL/hour to 0.4 +/- 1.9 ng/mL/hour (P < 0.001) and ln plasma aldosterone (PA) from 2.0 +/- 0.8 ng/dL to 2.3 +/- 0.8 ng/dL (P < 0.005) and fall in ln brain natriuretic peptide (BNP) from 4.3 +/- 0.9 pg/mL to 3.7 +/- 1.0 pg/mL (P < 0.01) were seen at 1 week. Despite a trend toward restoration of TBW, changes in hormonal biomarkers were maintained at 3 weeks. Over these 3 weeks, furosemide reduced 24-hour ambulatory blood pressure from 147 +/- 17/78 +/- 11 mm Hg to 138 +/- 21/74 +/- 12 mm Hg (P = 0.021) and torsemide reduced it from 143 +/- 18/75 +/- 10 mm Hg to 133 +/- 19/71 +/- 10 mm Hg (P = 0.007). CONCLUSION: Patients with CKD have elevated extracellular fluid volume that can be corrected acutely with loop diuretics. Persistent diuretic use results in dynamic changes in ECW and other body fluid compartments that translate into chronic blood pressure reduction. PMID- 14531811 TI - Protein restriction and AST-120 improve lipoprotein lipase and VLDL receptor in focal glomerulosclerosis. AB - BACKGROUND: Imai rats exhibit spontaneous focal glomerulosclerosis (FGS) with progressive proteinuria and hyperlipidemia leading to renal insufficiency by age 34 weeks. Recently, we reported marked down-regulations of skeletal muscle and adipose tissue lipoprotein lipase (LPL) and very low-density lipoprotein (VLDL) receptor in male Imai rats at 32 weeks of age. Dietary protein restriction and oral adsorbent AST-120 (AST) have been shown to slow progression of renal disease and attenuate hyperlipidemia in the Imai rats. This study tested the hypothesis that amelioration of proteinuria by protein restriction or use of oral adsorbent AST-120 beginning at 10 weeks of age may improve renal disease and LPL and VLDL receptor deficiencies in Imai rats. METHODS: Ten-week-old male Imai rats were randomly assigned to those fed either a regular diet, low protein diet (LPD), or regular diet containing the adsorbent preparation, AST-120. Ten-week-old male Sprague-Dawley rats served as controls. The animals were observed for 24 weeks. Six rats were included in each group. All diets were prepared in powder form. RESULTS: The untreated 34-week-old Imai rats showed severe proteinuria, hypoalbuminemia, 50% reduction in creatinine clearance, hypercholesterolemia, hypertriglyceridemia, and elevated plasma VLDL concentration. This was associated with significant reductions in plasma post-heparin LPL activity, hepatic lipase activity, as well as adipose tissue and skeletal muscle immunodetectable LPL and VLDL receptor proteins. Protein restriction mitigated the decline in creatinine clearance, ameliorated proteinuria, hypoalbuminemia, hypertension, and hypercholesterolemia, lowered plasma VLDL, and improved plasma postheparin LPL activity, hepatic lipase activity, LPL, and VLDL receptor proteins in skeletal muscle and adipose tissue. Similar improvements were observed in all parameters with AST administration. CONCLUSION: Moderate protein restriction and use of oral adsorbent can slow progression of renal disease and, thereby, ameliorate LPL, hepatic lipase, and VLDL receptor deficiencies and the associated hyperlipidemia in rats with spontaneous FGS. PMID- 14531812 TI - Epidermal basement membrane alpha 5(IV) expression in females with Alport syndrome and severity of renal disease. AB - BACKGROUND: X-linked Alport syndrome is a progressive nephritis caused by mutations of the COL4A5 gene. This gene encodes the collagen alpha 5(IV) chain, which is abnormally distributed in the glomerular basement membrane (GBM) and epidermal basement membrane (EBM). It has been reported a negative correlation between alpha 5(IV) chain distribution in EBM and the degree of proteinuria in heterozygous females with Alport syndrome. METHODS: In the present study, we evaluated the distribution of the alpha 5(IV) chain in the EBM and the degree of proteinuria in 22 females with X-linked Alport syndrome. The distribution of the cutaneous alpha 5(IV) chain was measured by a confocal laser microscope using an anti-alpha 5(IV) monoclonal antibody. The expression ratio of alpha 5(IV) distribution was quantified dividing the extension of the positive signal and the maximal extension of the specimen. Urinary protein excretion was expressed as urinary protein over urinary creatinine ratio. RESULTS: Proteinuria was present in five of the 22 patients. In two patients with proteinuria, alpha 5(IV)chain was normally distributed; in the remaining three, the expression ratio of alpha 5(IV)chain was 35%, 47%, and 48%. Of the 17 patients without proteinuria, two displayed a complete absence of the alpha 5(IV) chain in EBM, five displayed a normal staining, and the remaining 10 had an expression ratio between 18% and 65%. CONCLUSION: Our data suggest that there is no correlation between the severity of the glomerular involvement (expressed by proteinuria) and the staining of the alpha 5 chain in the EBM in females with X-linked Alport syndrome. PMID- 14531813 TI - The role of parental hypertension in the frequency and age of diagnosis of hypertension in offspring with autosomal-dominant polycystic kidney disease. AB - BACKGROUND: Hypertension in autosomal-dominant polycystic kidney disease (ADPKD) patients is associated with more rapid progression of renal disease and a high incidence of left ventricular hypertrophy (LVH). The present study was undertaken to examine the role of parental hypertension in the occurrence of hypertension in 475 ADPKD offspring. METHODS: Adult subjects participating in an ongoing study of the natural history of ADPKD were included in the analysis if they were diagnosed with ADPKD, had a known affected parent, and knew the hypertensive status of both parents. RESULTS: When the affected parent was hypertensive, the ADPKD male (82% versus 62%, P < 0.05) and female (61% versus 37%, P < 0.005) offspring had a significantly higher frequency of hypertension than when the ADPKD-affected parent was normotensive. The median age of diagnosis of hypertension was also significantly earlier in both male (33 years versus 40 years, P < 0.05) and female (38 years versus 50 years, P < 0.05) ADPKD patients when their affected parents were hypertensive as compared with normotensive. These effects of hypertension in the affected parent on hypertension in the ADPKD offspring were independent of age, renal volume, and renal function in the offspring. Hypertension in unaffected parents also increased the frequency of hypertension in the ADPKD female (69% versus 53%, P < 0.01), but not male (89% versus 77%, NS) subjects. CONCLUSION: The results indicate that parental hypertension influences the frequency of hypertension in ADPKD patients. PMID- 14531814 TI - Disease progression and outcomes in chronic kidney disease and renal transplantation. AB - BACKGROUND: It is unknown whether renal transplant recipients (RTR) have better outcomes and disease progression rates compared to patients with chronic kidney disease (CKD) when matched for the level of kidney function. METHODS: We analyzed data on 1762 patients with CKD (N = 872) and RTR (N = 890) over 16 years, applying the new Kidney/Disease Outcomes Quality Initiative (K/DOQI) staging system for CKD in a single center retrospective study. Patients were further divided based their native kidney disease. We determined disease progression by the slope of creatinine clearance decline and patient and kidney survival rates adjusted for age, gender and stage of kidney function, using Cox proportional hazards models. RESULTS: The overall rate of creatinine clearance decline in patients with CKD was -6.6 +/- 8.7 mL/min/year compared to -1.9 +/- 4.7 mL/min/year in RTR (P < 0.0001). The rate of decline per stage of CKD was also significantly lower in RTR. Whereas overall kidney survival was higher in RTR compared to patients with CKD (49.6% vs. 17.2%, respectively, P < 0.001), patient survival was not statistically different between the two groups (74.7% vs. 80.3%, respectively, P = 0.25). CONCLUSION: RTR had similar mortality rates compared to patients with CKD despite enjoying slower rates of disease progression and better kidney survival rates. These data suggest that RTR are a unique subset of patients with CKD whose comorbid conditions likely offset the potential benefits of slower rates of progression. PMID- 14531815 TI - End-stage renal disease in Scotland: outcomes and standards of care. AB - BACKGROUND: The number of patients starting renal replacement therapy (RRT) for end-stage renal disease (ESRD) in the United Kingdom rises annually. Patients are increasingly elderly with a greater prevalence of comorbid illness. Unadjusted survival, from the time of starting RRT, is not improving. The United Kingdom Renal Association has published recommended standards of treatment, which all United Kingdom nephrologists strive to attain. This study was devised to define the impact of attaining recommended treatment standards, adjusting for patient age and comorbid illnesses, upon survival on RRT in the United Kingdom population. METHODS: A prospective, registry based, observational study of all patients starting RRT in Scotland over a 1-year period, followed for the first 2 years of RRT. RESULTS: Of the 523 patients who were studied, 217 (41.5%) had died by 2 years of follow-up, 32% excluding deaths within the first 90 days. Age, comorbidity, weight when starting RRT, and attaining the recommended standards for albumin and hemoglobin had a significant impact upon survival. CONCLUSION: This study has emphasized the very high mortality of patients starting RRT in Scotland. By paying close attention to the attainment of recommended standards of care for patients with ESRD, it may be possible to improve upon current mortality figures. The monitoring of such success is only possible if correction is made for age and comorbidity. PMID- 14531816 TI - Urinary neopterin concentrations in patients with Balkan endemic nephropathy (BEN). AB - BACKGROUND: Balkan endemic nephropathy (BEN) is of great clinical importance in restricted areas of Bulgaria, Serbia, Croatia, Bosnia, Herzegovina, and Romania, since the etiology of BEN is still unknown. METHODS: In urine samples from 48 patients (41 females and 7 males, aged 65.6 +/- 6.87 years) with BEN living in an endemic area of Vratza district, Bulgaria, neopterin concentrations were measured by high-pressure liquid chromatography (HPLC) and compared with other clinical and laboratory investigations, including creatinine, hemoglobin, and erythrocyte sedimentation rates (ESRs). RESULTS: Urinary neopterin concentrations were 263 +/ 128 (mean +/- SD; range, 78 to 786 micromol/mol creatinine), 24 (50%) of BEN patients presented with increased concentrations as compared to the established normal ranges. Average ESRs were increased (1 hour, 29.0 +/- 14.7 mm/hour) and hemoglobin was decreased (109.3 +/- 16.4 g/L). Hemoglobin correlated inversely with ESRs (rs = -0.787 and -0.780) and creatinine concentrations (r = -0.690, all P < 0.001), but not with neopterin concentrations. Neopterin concentrations also did not correlate with serum creatinine levels. There existed an age relationship of ESR, creatinine, and hemoglobin, but not of neopterin. Neopterin concentrations were slightly lower in five females with low titers of antibodies against local B1 hantavirus strain (P < 0.05). CONCLUSION: The findings can support an immune-mediated inflammatory process in the pathogenesis of BEN only in a subgroup of patients. PMID- 14531817 TI - Supplemented very low protein diet ameliorates responsiveness to erythropoietin in chronic renal failure. AB - BACKGROUND: The aim of this study was to evaluate the relationship between uremic state and erythropoiesis in patients with predialytic chronic renal failure (CRF). METHODS: We monitored for 2 years the erythropoietin (EPO) requirement in patients with advanced CRF (creatinine clearance < or =25 mL/min), randomized to either low protein diet (LPD) group (0.6 g/kg body weight/day, N = 10) or very low protein diet (VLPD) group (0.3 g/kg body weight/day, N = 10) supplemented with a mixture of ketoanalogs and essential amino acids, both kept at target hemoglobin levels. RESULTS: The achieved protein intake after 6 months was 0.79 +/- 0.02 g/kg body weight/day and 0.50 +/- 0.02 g/kg body weight/day in LPD and VLPD, respectively; such a difference was maintained up to the end of follow up. The final hemoglobin values did not differ from the basal values in either group (11.5 +/- 0.2 g/dL and 11.5 +/- 0.3 g/dL). EPO dose, that was similar at baseline (62.4 +/- 9.6 UI/kg body weight/week and 61.8 +/- 8.8 UI/kg body weight/week subcutaneously), remained unchanged in LPD but progressively decreased in VLPD down to the final value of 41.2 +/- 7.0 UI/kg body weight/week (P < 0.0001 vs. basal and LPD). VLPD was associated with a decrease of urinary excretion and serum levels of urea nitrogen and phosphate; however, EPO requirement was not correlated with the changes of these parameters. On the contrary, the variation of EPO dose directly correlated with the modification of parathyroid hormone (PTH) levels, that diminished from 229 +/- 55 pg/mL to 118 +/- 16 pg/mL (P < 0.0001) in VLPD and did not change in LPD. CONCLUSION: In patients with advanced CRF, an effective decrease of protein intake of 0.3 g/kg body weight/day induces a reduction of about 35% of the EPO dose required to maintain the target hemoglobin levels. This effect appears dependent on the correction of a moderate secondary hyperparathyroidism. PMID- 14531818 TI - Hepatic lipase mutation may reduce vascular disease prevalence in hemodialysis patients with high CETP levels. AB - BACKGROUND: Uremic dyslipidemia characterized by reduced high-density lipoprotein (HDL) cholesterol levels is one of the major contributors to the high incidence of cardiovascular disease in hemodialysis patients. Hepatic lipase (HL), together with cholesteryl ester transfer protein (CETP), may not only promote reverse cholesterol transport but also enhance production of small, dense, more atherogenic low-density lipoprotein (LDL). A common C-514T mutation of the promoter region of the HL gene reportedly increases HDL cholesterol levels. However, whether the HL mutation is antiatherogenic or proatherogenic has remained unknown in uremic patients and the general population. METHODS: We investigated the influence of the mutation and its interaction with CETP on HDL cholesterol levels and the apparent atherosclerotic complications in 183 hemodialysis patients aged over 30 years who had received no antilipemic drugs. RESULTS: In patients with CETP levels > or =2.2 microg/mL [high CETP (HCT) group, N = 97], subjects with the TT genotype had a significantly higher level of HDL cholesterol than those without TT genotype (56.8 +/- 15.9 mg/dL vs. 45.7 +/- 13.4 mg/dL, P < 0.001), but not in patients with CETP levels <2.2 microg/mL [low CETP (LCT) group]. Multiple linear regression analysis showed that the TT genotype was a major independent positive determinant for HDL cholesterol levels in the HCT not LCT group. Among the HCT group patients, subjects with the TT genotype (N = 25) had a tendency toward lower prevalence of vascular disease than those without TT genotype (N = 72) (4.0% vs. 22.2%, P < 0.07). In this subgroup, TT genotype had an independent odds ratio of 0.041 (95% CI 0.002 to 0.75, P < 0.05) after adjusting for other risk factors. CONCLUSION: The TT genotype of HL mutation may serve as a protective factor against vascular disease by increasing HDL cholesterol levels in hemodialysis patients with higher CETP levels. PMID- 14531819 TI - Body size and outcomes on peritoneal dialysis in the United States. AB - BACKGROUND: Being overweight is often cited as a relative contraindication to peritoneal dialysis. Our primary objective was to determine whether actual mortality rates support this opinion. METHODS: Retrospective cohort study of United States Medicare patients initiating dialysis between 1995 and 2000 (N = 418,021; 11% peritoneal dialysis). RESULTS: Seven percent were underweight [body mass index (BMI) < 18.5 kg/m2], 27% were overweight (BMI 25.0 to 29.9 kg/m2), and 23% were obese (BMI> 29.9 kg/m2) at dialysis initiation. Compared to those with normal BMI, the adjusted odds of initiating peritoneal dialysis were 0.70 (P < 0.05) in underweight, 1.12 (P < 0.05) in overweight, and 0.87 (P < 0.05) in obese subjects. Among peritoneal dialysis patients, adjusted mortality hazard ratios in the first, second, and third year were 1.45 (P < 0.05), 1.28 (P < 0.05), and 1.17 for the underweight, respectively; 0.84 (P < 0.05), 0.89 (P < 0.05), and 0.98 for the overweight, respectively; and 0.89 (P < 0.05), 0.99, and 1.00 for the obese, respectively. Apart from higher third-year mortality in the obese, associations were similar after censoring at a switch to hemodialysis. For transplantation, the corresponding results were 0.76 (P < 0.05), 0.90 (P < 0.05), and 0.88 for the underweight, respectively; 0.95, 1.06, and 0.93 for the overweight, respectively; and 0.62 (P < 0.05), 0.68, and 0.71 for the obese, respectively. For switching to hemodialysis, hazards ratios were 0.92, 0.97, and 0.80 for the underweight, respectively; 1.07, 1.11 (P < 0.05), and 1.03 for the overweight, respectively; and 1.28 (P < 0.05), 1.29 (P < 0.05), and 1.36 (P < 0.05) for the obese, respectively. CONCLUSION: Although less likely to initiate peritoneal dialysis, overweight and obese peritoneal dialysis patients have longer survival than those with lower BMI, not adequately explained by lower transplantation and technique survival rates. PMID- 14531820 TI - Expression of urokinase plasminogen activator and its receptor during acute renal allograft rejection. AB - BACKGROUND: In inflammation, urokinase plasminogen activator (uPA) and its receptor (uPAR) play an important role in fibrinolysis and in activation and chemotaxis of neutrophils and lymphocytes. Moreover, the uPA/uPAR system is involved in processes that affect turnover of the extracellular matrix (ECM). The aim of this study was to determine the local and systemic release of uPAR, and the expression of uPA and uPAR in renal tissues during acute renal allograft rejection. METHODS: Blood, urine, and tissue samples were collected from 33 patients diagnosed with acute allograft rejection and from 14 transplant patients without rejection. From 10 healthy volunteers, blood and urine were collected as a control. In urine and blood samples, the levels of uPAR were determined by enzyme-linked immunosorbent assay (ELISA). Immunostaining and in situ hybridization for uPA and uPAR were performed on renal biopsies. RESULTS: uPAR was detectable at low levels in serum and urine of healthy volunteers and was increased in nonrejecting allograft recipients. Serum and urine levels of uPAR were higher in transplant recipients with rejection compared to nonrejectors. The urine and serum levels of uPAR correlated with the renal function. Immunostaining and in situ hybridization showed an up-regulation of both uPA and uPAR in rejection biopsies. Nonrejected grafts displayed no expression of uPA and uPAR by immunostaining, or of uPAR by in situ hybridization. uPA was detected in a limited number of tubular epithelial cells by in situ hybridization. During rejection, lymphocytes as well as tubular epithelial cells showed uPA and uPAR expression. In the vascular types of rejection, strong expression of uPA was also seen in the entire vessel wall, while uPAR was expressed by the endothelium. CONCLUSION: This study shows that (1) uPA and uPAR are up-regulated during acute renal allograft rejection; (2) uPAR levels in urine and serum correlate with serum creatinine levels, and (3) uPA and uPAR are produced by inflammatory cells, tubular epithelium, and vascular endothelium during acute renal allograft rejection. PMID- 14531821 TI - Glucose degradation products (GDP) retard remesothelialization independently of D glucose concentration. AB - BACKGROUND: Glucose degradation products (GDP) present in heat-sterilized dialysis fluids are thought to contribute to cellular dysfunction and membrane damage during peritoneal dialysis. To examine the effects of specific GDP on the remesothelialization process, the impact of conventional and low GDP peritoneal dialysis solutions, D-glucose, and individual GDP in a scratch-wounding model was assessed. METHODS: Scratch (0.5 to 0.6 mm)-wounded human peritoneal mesothelial cells (HPMC) were treated, at pH 7.4, with either (1) control medium (M199), (2) laboratory-prepared heat or filter-sterilized solutions, (3) 10% to 80% vol/vol solution of Gambrosol or Gambrosol-trio (1.5% and 4.0% glucose), (4) D-glucose (5 to 80 mmol/L), or (5) individual or combined GDP [acetaldehyde, formaldehyde, glyoxal, methylglyoxal, 3-deoxyglucosone (3-DG), 5-hydroxy methylfufural (5-HMF), or 3,4-di-deoxyglucosone-3-ene (3,4-DGE)]. Wound closure was recorded by time lapse photomicroscopy. RESULTS: In untreated HPMC, the rate of wound closure was linear and the process was complete by 18.4 +/- 3.6 hours (N = 16). In wounded HPMC exposed to dilutions of heat-sterilized but not filtered laboratory solutions (1.5% or 4.0% glucose, pH 7.4), remesothelialization was significantly retarded (P = 0.04 and P = 0.009 vs. M199, respectively). In Gambrosol, remesothelialization was significantly retarded in both 1.5% and 4.0% solutions. In contrast in Gambrosol-trio-treated HPMC, this rate was not significantly reduced in either 1.5% or 4.0% glucose peritoneal dialysis fluids. Remesothelialization was dose-dependently retarded in HPMC exposed to 3,4-DGE (>10 microl/L), formaldehyde (>5 micromol/L) but not by exposure to the other GDP tested even at 5 times the concentration present in low glucose solutions. The rate of remesothelialization was not significantly altered by exposure to D glucose concentrations up to 80 mmol/L. CONCLUSION: These data identify that the formaldehyde and 3,4-DGE present in heat-sterilized peritoneal dialysis solutions are important in reducing mesothelial cell regeneration. Specifically targeting their removal may have major benefits in preserving the mesothelium during long term peritoneal dialysis. PMID- 14531822 TI - Dynamics of PTH secretion in hemodialysis patients as determined by the intact and whole PTH assays. AB - BACKGROUND: Renal hyperparathyroidism is assessed by measurement of parathyroid hormone (PTH) levels. The intact PTH assay (I-PTH) not only reacts with 1-84 PTH but also with large, truncated fragments of non-1-84 PTH. Because the whole PTH assay (W-PTH) is specific for 1-84 PTH, non-1-84 PTH is determined by subtracting W-PTH from I-PTH values. These large circulating PTH fragments may exert a hypocalcemic effect by contributing to skeletal resistance to 1-84 PTH. METHODS: The dynamic secretion of both 1-84 PTH and non-1-84 PTH was evaluated during the induction of hypo- and hypercalcemia in eight hemodialysis patients. RESULTS: The basal ionized calcium concentration was 1.23 +/- 0.03 mmol/L at which time I-PTH, W-PTH, and non-1-84 PTH values were 276 +/- 78 pg/mL, 164 +/- 48 pg/mL, and 102 +/- 28 pg/mL, respectively. The induction of hypo- and hypercalcemic changes resulted in a sigmoidal response for all three PTH moieties, I-PTH, W-PTH, and non-1-84 PTH. During hypocalcemia, maximal values of W-PTH were greater than those of non-1-84 PTH. But during hypercalcemia, minimal values of W-PTH and non 1-84 PTH were similar. Neither the set points nor the basal/maximal ratios for W PTH, I-PTH, and non-1-84 PTH were different. At the baseline ionized calcium concentration, the W-PTH (1-84 PTH)/non-1-84 PTH ratio was 1.53 +/- 0.15. Changes in ionized calcium resulted in a sigmoidal relationship with hypocalcemia, increasing this ratio to a maximum of 2.01 +/- 0.30 and hypercalcemia decreasing this ratio to a minimum of 1.18 +/- 0.15 (P < 0.01 vs baseline for both hypo- and hypercalcemia). CONCLUSION: Although acute changes in serum calcium produce similar secretory responses in 1-84 PTH and non-1-84 PTH, the secretory responses are not proportional for these PTH moieties. Changes in the serum calcium concentration modulate the ratio of 1-84 PTH/non-1-84 PTH in a sigmoidal pattern with hypocalcemia maximizing this ratio. Whether changes in the 1-84 PTH/non-1-84 PTH ratio specifically modulate the calcemic action and other biologic effects of 1-84 PTH remain to be determined. PMID- 14531823 TI - Relationship between duration of brain death and hemodynamic (in)stability on progressive dysfunction and increased immunologic activation of donor kidneys. AB - BACKGROUND: Consistent difference in graft survival after renal transplantation has been shown when cadaveric transplants are compared to the living related donor situation, in favor of the latter. Recently, evidence has been put forward that brain death has significant effects on the donor organ quality. In this study, we aimed to assess the relation between brain death-induced hemodynamic instability in combination with the duration of brain death on the function and immunogenicity status of potential donor kidneys. METHODS: In Wistar rats, short term (1 hour) or long-term (6 hours) brain death in the presence or absence of hemodynamic stability was applied. Sham-operated rats served as controls (1 hour and 6 hours). Organ function was studied by monitoring serum creatinine, lactate dehydrogenase (LDH), lactate, and total protein content. Expression of cell adhesion molecules [intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1)] and the influx of leukocytes in the kidney assessed the immunologic status of the kidney. RESULTS: Progressive organ dysfunction was most pronounced in hemodynamically unstable brain-dead donors reflected by increased serum creatinine levels. Regardless of hemodynamic status, a progressive inflammatory activation by cell adhesion molecule expression and an influx of leukocytes could be observed in kidneys of brain-dead rats compared with nonbrain-dead controls. CONCLUSION: Brain death causes progressive kidney dysfunction. Also, inflammatory responses reflecting tissue injury are caused by brain death. When hemodynamic instability in the brain-dead donor is not corrected, kidney dysfunction is enhanced and immune activation occurs faster and is more profound. The observed changes may predispose the graft for additional ischemia/reperfusion injury during the transplant process and hence accelerate rejection of the graft after transplantation. PMID- 14531824 TI - Aristolochic acid nephropathy and the peritoneum: Functional, structural, and molecular studies. AB - BACKGROUND: Aristolochic acid nephropathy (AAN) is a rapidly progressive interstitial nephropathy linked to the exposure to aristolochic acid (AA) and characterized by extensive fibrosis and urothelial atypia. Although the fibrotic process has been documented in extrarenal tissues, the involvement of the peritoneum, as well as the efficacy of peritoneal dialysis in AAN patients, remain uncertain. METHODS: The structure of the peritoneal membrane and the expression of basic fibroblast growth factor (bFGF), collagen type III, endothelial nitric oxide synthase (eNOS), and aquaporin-1 (AQP1) were investigated in peritoneal biopsies from an index AAN patient, four other AAN patients, four regular peritoneal dialysis patients, and two controls. Similar methods were used to investigate a rabbit model of AAN after intraperitoneal exposure to high-dose AA. AA-DNA adducts were screened by 32P-postlabeling analysis. RESULTS: The AAN patients had renal failure, renal fibrosis, and urothelial atypia. The peritoneum of AAN patients had a normal structure, lacked cellular atypia, and, in comparison with regular peritoneal dialysis patients and controls, did not show abnormal regulation of fibrotic and endothelial markers. Furthermore, specific AA-DNA adducts were not identified in the peritoneum of AAN patients. In contrast, AA-DNA adducts were detected in peritoneal and kidney tissues of all exposed rabbits, and one of them developed a malignant mesothelioma. CONCLUSION: These data demonstrate the lack of fibrotic and vascular alterations and the absence of cellular atypia in the peritoneum from AAN patients. Thus, peritoneal dialysis should not be discouraged in these patients. Nevertheless, studies in a rabbit model of high-dose AA exposure may suggest a potential risk of peritoneal malignancy. PMID- 14531825 TI - Epidemiology of vascular access in the Australian hemodialysis population. AB - BACKGROUND: A number of demographic and comorbid factors have been demonstrated to be associated with the placement of arteriovenous grafts (AVG) and central venous catheters (CVC) as opposed to native arteriovenous fistulas (AVF). However, no data are available regarding these factors in a hemodialysis population where AVF utilization is high. METHODS: All adult patients on hemodialysis on September 30, 2001 in Australia were included in the study. Vascular access was recorded as AVF, AVG, or CVC. Patients were separated into incident (<150 days since first dialysis) and prevalent cohorts (> or =150 days). Multinomial logistic regression was used to assess factors associated with AVG and CVC use. RESULTS: Of the 4968 patients who were studied, 877(17%) were classed as incident and the remainder prevalent. AVF were present in 61% versus 77%, AVG were present in 11% versus 19%, and CVC were present in 28% versus 4% in the incident and prevalent cohorts, respectively (all P < 0.001). After adjustment for confounding factors, age and female gender were associated with an increased frequency of AVG in both cohorts. In addition, type I diabetes mellitus was associated with increased frequency of AVG use in the incident cohort, whereas body mass index (BMI) > or =30 kg/m2 and peripheral vascular and cerebrovascular disease were significant in the prevalent group. For CVC, female gender, type I and II diabetes mellitus and late referral were associated with increased frequency in the incident cohort, while females, cigarette smoking, and peripheral vascular disease were predictive in the prevalent group. Significant variations in access type were also seen depending on geographic location. CONCLUSION: Certain patient characteristics such as age and female gender, but not type II diabetes mellitus, remain significantly associated with AVG and catheter use despite the high prevalence of AVF use in Australia. However, the significant variation in risk by geographic location suggests more attention needs to be paid to physician practice patterns to increase AVF utilization rates. PMID- 14531826 TI - Health-related quality of life among dialysis patients on three continents: the Dialysis Outcomes and Practice Patterns Study. AB - BACKGROUND: Assessing health-related quality of life (HRQOL) can provide information on the types and degrees of burdens that afflict patients with chronic medical conditions, including end-stage renal disease (ESRD). Several studies have shown important international differences among ESRD patients treated with hemodialysis, but no studies have compared these patients' HRQOL. Our goal was to document international differences in HRQOL among dialysis patients and to identify possible explanations of those differences. METHODS: We examined data from the Dialysis Outcomes and Practice Patterns Study (DOPPS), a prospective, observational, international study of hemodialysis patients. We performed a cross-sectional analysis of DOPPS data from the United States, five countries in Europe (France, Germany, Italy, Spain, and the United Kingdom), and Japan. Linear mixed models were used to analyze differences in HRQOL, using the KDQOL-SFTM. Norm-based scores were used to minimize cultural response bias. Linear regression analysis was used to adjust for confounding factors. Other variables included demographic variables, comorbidities, primary cause of ESRD, complications of ESRD and treatment, and socioeconomic status. RESULTS: In all generic HRQOL subscales, patients on all three continents had much lower scores than their respective population norm values. Patients in the United States had the highest scores on the mental health subscale and the highest mental component summary scores. Japanese patients reported better physical functioning than did patients in the United States or Europe, but they also reported the greatest burden of kidney disease. Overall, these differences remained even after adjusting for possible confounders. CONCLUSION: On all three continents, ESRD and hemodialysis profoundly affect HRQOL. In the United States, the effects on mental health are smaller than in other countries. Japanese hemodialysis patients perceived that their kidney disease imposes a greater burden, but their physical functioning was significantly higher. Different distributions of socioeconomic factors and major comorbid conditions could explain little of this difference in physical functioning. Other possible factors, such as quality of dialysis and related health care, deserve careful study. PMID- 14531827 TI - Measuring transport of water across the peritoneal membrane. AB - INTRODUCTION: Mechanisms of water flow across the peritoneal membrane include diffusion, convection, and reabsorption. OBJECTIVES: To understand these processes more clearly we have developed a method to measure transport of water across the peritoneal membrane. METHODS: An artificial gradient of deuterated water (HDO) between blood and dialysate compartments was created in five subjects who took 0.3g per kg of body weight of D2O, which was allowed to equilibrate with total body water. During a test dwell (2 L, bicarbonate:lactate buffer, 1.36% glucose to minimize convection), frequent dialysate samples were drawn to determine the abundance of deuterium and other solutes and to calculate their time constants. Dialysate deuterium abundance was measured using flowing afterglow mass spectrometry (FA-MS). The method was combined with 125iodine labeled albumin (RISA) to enable simultaneous estimates of intraperitoneal volume and thus calculation of the mass transfer area coefficient (MTAC) for small solutes using the Garred equation. RESULTS: The appearance of HDO in dialysate in four subjects is described by a single exponential fit with residuals of <1%, similar to method precision. In a fifth subject, the resolution of this method demonstrated that the best fit was a double exponential. When compared to other solutes, the time constant for water was as predicted by its molecular weight, with a MTAC of 38.7 +/- 4.4 mL/min. Total body water could also be estimated from the equilibrated dialysate deuterium abundance, with repeat estimates within 0.5%. CONCLUSION: Transport of water across the peritoneum can be measured with remarkable accuracy and when combined with an intraperitoneal volume estimation can be used to determine mass transfer. In conditions of low convection, the relative rate of deuterium appearance and mass transfer compared to other solutes suggests that water diffuses predominantly through the intercellular small pores. PMID- 14531828 TI - Scaffolds: Orchestrating proteins to achieve concerted function. PMID- 14531829 TI - Lipoxygenases in renal injury--loading the matrix. PMID- 14531830 TI - Supplementary data to the study of ESRD Map in Japan. PMID- 14531831 TI - Stopping progression in familial juvenile hyperuricemic nephropathy with benzbromarone? PMID- 14531832 TI - Submaximal dose of trandolapril in the COOPERATE trial? PMID- 14531834 TI - Inverse association between homocysteine and vascular endothelial growth factor receptor 2 serum levels in hemodialyzed and kidney transplanted patients. PMID- 14531835 TI - End-organ susceptibility as a determinant of renal disease in hypertension. PMID- 14531836 TI - Presentation of the 2003 Jean Hamburger Award to J. Stewart Cameron. PMID- 14531837 TI - Acceptance of the 2003 Jean Hamburger Award. PMID- 14531839 TI - Acceptance of the 2003 Jean Hamburger Award. PMID- 14531838 TI - Presentation of the 2003 Jean Hamburger Award to Robert W. Schrier. PMID- 14531840 TI - Presentation of the 2003 A.N. Richards Award to Marilyn Farquhar. PMID- 14531841 TI - Acceptance of the 2003 A.N. Richards Award. PMID- 14531842 TI - N-terminal modifications leading to peptide ORL1 partial agonists and antagonists. AB - Nociceptin/Orphanin FQ (N/OFQ) is a 17 amino acid peptide that is the endogenous ligand for the G protein-coupled receptor (opioid receptor like 1, ORL1), a member of the opioid receptor family. Although it is clear that this receptor system is involved in a variety of physiological functions, including analgesia, the precise actions of N/OFQ remain largely uncharacterized. One reason for this has been limited high affinity ligands to ORL1, and particularly the lack of availability of useful specific antagonists. Herein we describe the pharmacological activity of a series of N-terminally modified hexapeptides with high affinity for ORL1. These compounds were tested for binding affinity using [3H]N/OFQ binding to human ORL1 in CHO cells, and functional activity by measuring stimulation of [35S]GTPgammaS binding in CHO cell membranes. The N terminal modifications have produced compounds that maintained very high receptor affinity, but led to significant changes in intrinsic activity. One compound, pentanoyl-RYYRWR-NH2, with barely measurable agonist activity was tested in vivo. It was found to possess modest analgesic activity, but it was unable to block the morphine modulatory activity of N/OFQ. PMID- 14531843 TI - Extensive structure-activity studies of lactam derivatives of MT-II and SHU-9119: their activity and selectivity at human melanocortin receptors 3, 4, and 5. AB - The melanocortin system is involved in the regulation of several diverse physiologic pathways. Recently we have demonstrated that replacing His6 by Pro6 in the well-known antagonist SHU-9119 resulted in a potent agonist at the hMC5R (EC50 = 0.072 nm) with full antagonist activity at the hMC3R and the hMC4R. We have designed, synthesized, and pharmacologically characterized a series of peptide analogs of MT-II and SHU-9119 at the human melanocortin receptors MC3R, MC4R and MC5R. All these peptides were modified at position 6 with a Pro instead of a His residue. In this study, we have identified new scaffolds which are antagonists at the hMC4R and hMC3R. Additionally, we have discovered a new selective agonist at the hMC4R, Ac-Nle-c[Asp-Pro-D-Phe-Arg-Trp-Lys]-Pro-Val-NH2 (6, PG-931) which will be useful in further biologic investigations of the hMC4R. PG-931 was about 100-fold more selective for the hMC4R vs. the hMC3R (IC50 = 0.58 and 55 nm, respectively). Some of these new analogs have exceptional biologic potencies at the hMC5R and will be useful in further efforts to differentiate the substructural features responsible for selectivity at the hMC3R, hMC4R, and hMC5R. PMID- 14531844 TI - Isolation of peptides of the brevinin-1 family with potent candidacidal activity from the skin secretions of the frog Rana boylii. AB - The emergence of strains of the human pathogen Candida albicans with resistance to commonly used antibiotics has necessitated a search for new types of antifungal agents. Six peptides with antimicrobial activity were isolated from norepinephrine-stimulated skin secretions from the foothill yellow-legged frog Rana boylii. Brevinin-1BYa (FLPILASLAA10KFGPKLF CLV20TKKC) was particularly potent against C. albicans [minimal inhibitory concentration (MIC) = 3 microm] and also active against Escherichia coli (MIC = 17 microm) and Staphylococcus aureus (MIC = 2 microm), but its therapeutic potential for systemic use is limited by its strong hemolytic activity (HC50 = 4 microm). The single amino acid substitution (Phe12 --> Leu) in brevinin-1BYb resulted in a fourfold lower potency against C. albicans and the additional amino acid substitutions (Lys11 - > Thr, Phe17 --> Leu and Val20 --> Ile) in brevinin-1BYc resulted in a ninefold decrease in activity. Two members of the ranatuerin-2 family and one member of the temporin family were also isolated from the secretions but showed relatively low potency against the three microorganisms tested. PMID- 14531845 TI - Using protein-based motifs to stabilize peptides. AB - While the use of synthetically derived novel inhibitor peptides as a source of new therapeutics for medicine remains incredibly promising, there is a major problem with implementing this technology, as many synthetic peptides have proven to be unstable and are degraded by peptidases in the host cell. In this study, we have investigated methods by which peptides can be stabilized using protein-based motifs in order to prevent the action of peptidases. Using an in vivo approach our laboratory developed to screen for synthetic peptides which can inhibit the growth of Escherichia coli, we found that protecting the amino or carboxyl terminus of the peptides via fusion to the very stable Rop protein, or the incorporation of two proline residues, increased the frequency at which potent inhibitor peptides could be isolated. Using an in vitro degradation assay in which extracts from several different cell types were tested, we demonstrated that peptides stabilized with multiple proline residues were more resistant to degradation than peptides stabilized by amidation or acetylation, two approaches that are routinely utilized to improve the stability of peptide drugs. PMID- 14531846 TI - Tachyphylactic properties of angiotensin II analogs with bulky and hydrophobic substituents at the N-terminus. AB - Tachyphylaxis, defined as the acute loss of response of some smooth muscles upon repeated stimulations with angiotensin II (Ang II), has been shown to be dependent mainly on the N-terminal region of the ligand. To further study the structural requirements for the induction of tachyphylaxis we have synthesized Ang II analogs containing the bulky and very lipophilic substituents 9 fluorenylmethyloxycarbonyl (Fmoc) and 9-fluorenylmethyl ester (OFm) at the alpha amino (Nalpha-Fmoc-Ang II) or the beta-carboxyl ([Asp(OFm)1]-Ang II) groups of the Asp1 residue, respectively. In binding assays with Chinese hamster ovary cells transfected with the AT1 Ang II receptor, Nalpha-Fmoc-Ang II bound with high affinity, whereas [Asp(OFm)1]-Ang II showed lower affinity. In biological assays, these two analogs were full agonists and showed 30 and 3%, respectively, of the Ang II potency in contracting the guinea-pig ileum smooth muscle. The two analogs induced tachyphylaxis, in spite of the lack of a free amino group in Nalpha-Fmoc-Ang II. Thus, analogs with Fmoc- or OFm-type groups coupled to the Asp1 residue, whether at the amino or carboxyl functions, induce tachyphylaxis through an unreported mechanism. Based in these findings and those available from the literature, an alternate molecular interaction mode between Ang II N-terminal portion and the AT1 receptor is proposed to explain the tachyphylactic phenomenon. PMID- 14531847 TI - The role of documentation in making nursing work visible. PMID- 14531848 TI - Evidence-based practice: to be or not to be, this is the question! AB - Evidence-based nursing is the current fashion. It is being touted as the mechanism to achieve best practice in the clinical setting. But while evidence based practice (EBP) is being presented in the literature, discussed at nursing practice forums, and evidence-based centres of excellence have developed, there seems to be very little impact in the practice that nurses deliver on a daily basis. The case in point is the collection of vital signs. While not historically a nursing skill, over the last 60 years it has become an integral component of practice in the postoperative general surgical setting. The evidence to support these practices is scant. Policies and text purport traditional routine-regulated practice without substantive evidence to support their claims. These policies are being used to control rather than support EBP. In conjunction with the traditional practice of vital sign collection and the culture of the clinical settings, the policies are limiting opportunities for clinicians to make individual decisions about care delivery based on the unique needs of each patient. Rather than focusing on EBP as the solution to the development of best practice, is it not time to change the focus to real strategies that will assist in achieving best practice? These include the creation of rigorous relevant evidence, the valuing of clinical expertise and the changing of the cultures in which nurses develop and practice. PMID- 14531849 TI - Imbedding quality improvement into all aspects of nursing practice. AB - Influencing nurses to accept and implement best practice and continuous quality improvement into everyday nursing activities and to change practice is a significant challenge for any nursing service. Quality improvement is often considered by clinicians to be yet another task to be added to their already busy workloads. This paper describes the attempts of one large metropolitan tertiary teaching hospital to imbed quality improvement and best practice across the spectrum of its nursing service and to foster a culture of quality within clinical practice and documentation. It outlines the methods used by one hospital to enshrine best practice into all nursing activities and to minimize variations in practice throughout the hospital through the use of its nursing clinical information system. PMID- 14531850 TI - Evaluating the impact of a new high dependency unit. AB - The impact of a new high dependency unit (HDU) that was developed from an adjoining ward was evaluated from both an organizational and consumer perspective. Satisfaction of patients and relatives with nursing care on the ward of origin was measured before and after the development of the HDU, and nurses were surveyed regarding the impact of the change on nursing practices. Patients' perceptions of pain and pain management was compared between patients admitted to the ward prior to the development of the HDU, and a later group admitted to the HDU. Results showed that despite high satisfaction prior to the development of the HDU, satisfaction of patients and relatives improved on the ward after the development of the HDU. Nurses in both areas also described higher satisfaction levels after the change. Patients admitted to the HDU expressed higher levels of satisfaction with nurses' response to analgesia requests and clinical data showed a trend toward lower pain scores in this group compared with pre-HDU patients. The HDU was found to be effective in providing high level care and monitoring for a group of patients at risk of postoperative complications. It was also associated with additional benefits in the running of the adjacent ward. PMID- 14531851 TI - Quality improvement measures based on patient data: some psychometric issues. AB - Existing methods for handling patients' opinions as a basis for quality improvement measures tend to be too sophisticated for practical nursing and/or weak psychometrically. An Index of Measures was developed by combining patients' perceptions of actual care conditions with the subjective importance they ascribed to these conditions. Data from Swedish somatic inpatients (n = 4002) indicated that this index gave added value compared to patients' ratings of actual care conditions only. Interpretation of the Index of Measures focused on the proportion of patients who receive the value "deficiency." This value is obtained if a given patient has a subjective importance rating on a given item which is higher than the mean subjective importance score for the whole patient group on that item, as well as a perceived reality rating on the item in question which is lower than the mean perceived reality rating for the whole group on that item. Guidelines are suggested on when improvement actions are necessary and when they are not necessary. PMID- 14531853 TI - Agitated older patients: nurses' perceptions and reality. AB - Management of agitation is an increasingly common duty in the nursing care of older patients. Agitation describes a range of behavioural symptoms including aggressiveness and hyperactivity, and can be a sign of severe illness. In a series of focus groups in a large teaching hospital, nurses' feelings and actions in regard to nursing agitated older people were recorded and analysed. The results demonstrated that agitation is not well understood and is often not well managed. Despite recent efforts to provide more supportive staffing structures, nurses felt particularly stressed and unsupported in relation to this phenomenon. Recommendations are made for the instigation of enhanced education programmes as well as ongoing attention to improved workload initiatives. PMID- 14531852 TI - Adequacy of support for new graduates during their transition into the workplace: a Queensland, Australia study. AB - In 2001, a study into issues of concern to assistant-in-nursing, registered and enrolled nurse members of the Queensland Nurses Union was undertaken. Approximately equal numbers of nurses from each of the aged care, acute private and acute public sectors were surveyed. Overall, 1477 nurses responded, representing a response rate of 53%. This article focuses on one aspect of the study-the perceived adequacy of support offered to new nursing graduates as they exit university and begin their transition into the workplace. In particular, responses from nurses are compared with professional level or current role designation, age, time spent in the workplace and health sector. Considerable divergence of opinion among the respondents, particularly across designation, age and years of experience, was found within the three sectors. For example, in the public and private employment sectors, older and more experienced nurses were more likely to perceive the support for new nurses as adequate compared to younger and less experienced nurses. Additionally, in the acute private sector, the more senior the nurse the more likely the perception that there was adequate preparation for new graduates entering the workforce. PMID- 14531854 TI - Predictors of psychological well-being of nurses in Alexandria, Egypt. AB - As nursing has been previously identified as a stressful occupation, sources of job stress and levels of job satisfaction were extensively investigated. However, studies addressing issues of nurses' mental health are scarce. This work aims to assess psychological well-being of nurses in different job settings in Alexandria and to identify sociodemographic, psychosocial and workplace predictors. A total sample of 412 nurses represented nurses working in five different health organizations in Alexandria. A self-administered questionnaire was used to collect sociodemographic, occupational and health data, and the Standardized Arabic Version of General Health questionnaire (GHQ-30 items), Job Descriptive Index (JDI), and Social Support Scale (SSS) were also used. Results revealed that 21.67% of nurses recorded moderate to severe psychological symptoms on GHQ. Fewer years of experience, negative family and friend support, and negative total work satisfaction were found to be significant predictors of psychological ill health among nurses in a descending rank order. Implications for nursing intervention will be discussed. PMID- 14531855 TI - The spiritual dimensions of care in military nursing practice. AB - Spirituality has been the subject of numerous journal articles and books in recent years. Research into this topic has been conducted in many spheres of nursing practice with the notable exception of military nursing. This article goes a small way to addressing the apparent lack of research into spirituality in a military nursing setting by summarizing the findings of one study into this significant area of nursing care. The findings are derived from a mixed method quantitative/qualitative study of registered nurses in the Royal Australian Air Force. The major finding indicated that two distinct concepts of "family" define the way in which this small group of nurses perceive, assess and implement care for the spiritual needs of their patients. These concepts comprise a traditional family structure and an extended military family structure that includes the person's unit and comrades-in-arms. PMID- 14531856 TI - The importance of social and cultural factors to nursing status. AB - The objective of the authors was to examine nursing staff workload. The examination is based on the workload model of Robert Karasek and on the status integration theory of P. Gibbs and Walter T. Martin. As a means of examination, an open questionnaire consisting of 55 items and 76 variables was given to nurses working in different fields of health care, studying in higher education instead, or to persons with a degree in nursing (n = 465). The main concern of the authors was to attempt to identify the inconsistencies in nursing status which might come from limited decision-making freedom and high workplace requirements. In the process of statistical analysis of the data, considerable deviation was observed regarding some scales of the questionnaire. The authors emphasize that the reasons for the inconsistencies in nursing status might originate either from traditional cultural elements or from increased workload based on unclear competencies which are forced on the nurses. PMID- 14531857 TI - Transcription elongation factor S-II maintains transcriptional fidelity and confers oxidative stress resistance. AB - BACKGROUND: During transcription elongation, RNA polymerase II is arrested on the template when incorrect ribonucleotides are incorporated into the nascent transcripts. Transcription factor S-II enhances the excision of these mis incorporated nucleotides by RNA polymerase II and stimulates transcription elongation in vitro. This mechanism is considered to be transcriptional proof reading, but its physiological relevance remains unknown. RESULTS: We report that S-II contributes to the maintenance of transcriptional fidelity in vivo. We employed a genetic reporter assay utilizing a mutated lacZ gene from which active beta-galactosidase protein is expressed when mRNA proof-reading is compromised. In S-II-disrupted mutant yeasts, beta-galactosidase activity was ninefold higher than that in wild-type. The S-II mutant exhibited sensitivity to oxidants, which was suppressed by introduction of the S-II gene. The mutant S-II proteins, which are unable to stimulate transcription by RNA polymerase II in vitro, did not suppress the sensitivity of the mutants to oxidative stress or maintain transcriptional fidelity. CONCLUSION: These results suggest that S-II confers oxidative stress resistance by providing an mRNA proof-reading mechanism during transcription elongation. PMID- 14531858 TI - pH-dependent translocation of alpha-tocopherol transfer protein (alpha-TTP) between hepatic cytosol and late endosomes. AB - BACKGROUND: alpha-Tocopherol transfer protein (alpha-TTP), a member of the Sec14 protein family, plays an important role in transporting alpha-tocopherol, a major lipid-soluble anti-oxidant, in the cytosolic compartment of hepatocytes and is known as a product of the causative gene for familial isolated vitamin E deficiency. It has been shown that the secretion of hepatocyte alpha-tocopherol taken up with plasma lipoproteins is facilitated by alpha-TTP. To explore the mechanism of alpha-TTP mediated alpha-tocopherol secretion, we investigated drugs which may affect this secretion. RESULTS: We found that, in a hepatocyte cell culture system, intracellular alpha-tocopherol transport is impaired by chloroquine, an agent known for its function of elevating the pH in acidic compartments. Under chloroquine treatment, the diffuse cytosolic distribution of alpha-TTP changes to a punctate pattern. Double-staining experiments with endocytosis markers revealed that alpha-TTP accumulates transiently on the cytoplasmic surface of late endosomal membranes. This phenomenon is specific for hepatoma cell lines or primarily cultured hepatocytes. Other members of the Sec14 family, such as cellular retinaldehyde-binding protein (CRALBP) and supernatant protein factor (SPF), do not show this accumulation. Furthermore, we elucidate that the obligatory amino acid sequence for this function is located between amino acids 21 and 50, upstream of the N-terminal end of the lipid-binding domain. CONCLUSION: We hypothesize that a liver-specific target molecule for alpha-TTP exists on the late endosomal membrane surface. This transient binding may explain the mechanism of how alpha-tocopherol is transferred from late endosomes to cytosolic alpha-TTP. PMID- 14531859 TI - CspB and CspL, thermostable cold-shock proteins from Thermotoga maritima. AB - BACKGROUND: Cold-shock proteins (Csps) are important for cellular adaptation to low temperature. Csps help cells adapt to low-temperature growth through their RNA-binding and nucleic acid melting abilities, which lead to anti-termination of transcription. RESULTS: We studied the two most thermostable Csps known to date, TmCspB and TmCspL from Thermotoga maritima, a hyperthermophilic eubacterium for which no cold-shock response has been demonstrated so far. For comparison, we used a well-characterized Escherichia coli CspE protein. TmCspB and TmCspL are able to bind RNA at both low and high temperatures. They are also able to 'melt' nucleic acids secondary structures and as a result decrease E. coli RNA polymerase transcription termination in vivo and E. coli and T. maritima RNA polymerases transcription termination in vitro. Over-expression of TmCsps allowed E. coli cold-sensitive mutant cells to acclimate to the low temperatures of 15 degrees C. CONCLUSIONS: TmCspB and TmCspL (i) are able to perform essential functions of E. coli Csps in vitro and in vivo, 50-65 degrees C below the temperature optimum of T. maritima and (ii) can anti-terminate transcription by T. maritima RNA polymerase at 55 degrees C, the lower limit of temperature range for growth of T. maritima. We propose that the observed properties of TmCsps are physiologically relevant and that TmCsps are important for adaptation of T. maritima to physiologically low temperatures. PMID- 14531860 TI - Differential activities, subcellular distribution and tissue expression patterns of three members of Slingshot family phosphatases that dephosphorylate cofilin. AB - BACKGROUND: Cofilin, a key regulator of actin filament dynamics, is inactivated by phosphorylation at Ser-3 by LIM-kinases and is reactivated by dephosphorylation by a family of protein phosphatases, termed Slingshot (SSH). RESULTS: We have identified two novel isoforms of SSHs, termed SSH-2L and SSH-3L and characterized them in comparison with SSH-1L that was previously reported. SSH-1L and SSH-2L, but not SSH-3L, tightly bound to and co-localized with actin filaments. When expressed in cultured cells, SSH-1L, SSH-2L and SSH-3L decreased the level of Ser-3-phosphorylated cofilin (P-cofilin) in cells and suppressed LIM kinase-induced actin reorganization, although SSH-3L was less effective than SSH 1L and SSH-2L. In cell-free assays, SSH-1L and SSH-2L efficiently dephosphorylated P-cofilin, whereas SSH-3L did do so only weakly. Using deleted mutants of SSH-1L and SSH-2L, we found that the N-terminal and C-terminal extracatalytic regions are critical for cofilin-phosphatase and F-actin-binding activities, respectively. In situ hybridization analyses revealed characteristic patterns of expression of each of the mouse Ssh genes in both neuronal and non neuronal tissues; in particular, expression of Ssh-3 in epithelial tissues is evident. CONCLUSION: SSH-1L, SSH-2L and SSH-3L have the potential to dephosphorylate P-cofilin, but subcellular distribution, F-actin-binding activity, specific phosphatase activity and expression patterns significantly differ, which suggests that they have related but distinct functions in various cellular and developmental events. PMID- 14531862 TI - Abstracts of the annual meeting of the American Epilepsy Society. Boston, Massachusetts, USA. December 5-10, 2003. PMID- 14531861 TI - Negative regulation of Lyn protein-tyrosine kinase by c-Cbl ubiquitin-protein ligase in Fc epsilon RI-mediated mast cell activation. AB - BACKGROUND: Recent studies have demonstrated that c-Cbl functions as a ubiquitin protein ligase toward immune receptors and non-receptor protein-tyrosine kinase Syk by facilitating their ubiquitination and subsequent targeting to proteasomes. However, it was not clear whether Src family kinase Lyn is regulated by the Cbl family of ubiquitin-protein ligases. RESULTS: Aggregation of the high affinity IgE receptor (Fc epsilon RI) induces the rapid ubiquitination of Lyn in rat basophilic leukaemia RBL-2H3 cells. Treatment of cells with a proteasome inhibitor enhances the ubiquitination of Lyn. Stimulation of Fc epsilon RI results in the association of Lyn with c-Cbl and Cbl-b, both of which then become tyrosine phosphorylated. Co-transfection study shows that both c-Cbl and Cbl-b could induce the ubiquitination of activated Lyn in COS cells. Furthermore, over expression of membrane-anchored form of c-Cbl inhibits the Fc epsilon RI-mediated degranulation and cytokine gene production in RBL-2H3 cells by the down regulation of the kinase activity of Lyn through the enhanced ubiquitination. CONCLUSIONS: These results demonstrate that Lyn is down-regulated by c-Cbl mediated ubiquitination and subsequent degradation in proteasome after Fc epsilon RI stimulation in mast cells. Targeting of c-Cbl in the lipid raft results in the inhibition of Fc epsilon RI-mediated mast cell activation. PMID- 14531866 TI - A randomized comparison of an emollient containing skin-related lipids with a petrolatum-based emollient as adjunct in the treatment of chronic hand dermatitis. AB - Hand dermatitis is a multifactorial skin disorder in which skin barrier impairment is involved in the pathogenesis. The development of topical agents that improve skin barrier function is therefore a promising approach for the management of hand dermatitis. Topically applied lipids may interfere with skin barrier function, and emollients containing skin-related lipids have been suggested to facilitate repair of the skin barrier. However, evidence for the superiority of emollients containing skin-related lipids over the more traditional emollients is still lacking. The aim of this study was to compare an emollient containing skin-related lipids (Locobase Repair) with a traditional petrolatum-based emollient for the management of hand dermatitis. Adult males and females (n = 30) with mild to moderate chronic hand dermatitis were treated twice daily for 2 months either with an emollient containing skin-related lipids or with a pet.-based emollient. In the case of exacerbation, the patients of both treatment groups were allowed to use a mild corticosteroid according to instructions. Both treatment regimes significantly improved clinical signs of hand dermatitis as assessed by the investigator global assessment, hand eczema area and severity score. We did not observe significant differences in the improvement of clinical signs, itching, patients' assessment of efficacy, cosmetic acceptability or usage of topical corticosteroids between both treatment groups. In conclusion, this study confirms that the frequent use of emollients may be useful in the therapy of hand dermatitis. However, we could not demonstrate the superiority of this particular emollient containing skin-related lipids in patients with chronic hand dermatitis. PMID- 14531867 TI - Release of nickel ions from stainless steel alloys used in dental braces and their patch test reactivity in nickel-sensitive individuals. AB - Nickel ions leached in sufficient quantities from nickel-containing alloys may induce nickel sensitization or elicit allergic contact dermatitis. Nickel containing stainless steel alloys are generally considered safe for nickel sensitive individuals to use. The study summarized in this paper investigated 3 parameters. First, the release of nickel was estimated in artificial saliva and sweat from 4 different stainless steel alloys frequently used in dental braces. Second, in a pilot study, oral mucosa cells harvested from 3 dental patients before and after the attachment of dental braces were analysed for possible nickel content. Third, patch test reactivity of the 4 stainless steel alloys was tested on 31 nickel-sensitive subjects. All 4 stainless steel alloys released small amounts of nickel ions into artificial saliva (<0.13 micro g/cm2/week) and artificial sweat (<0.05 micro g/cm2/week), but no measurable amounts of nickel were found in any of the oral mucosa samples. None of the 31 nickel-sensitive subjects reacted to patch testing with the 4 stainless steel alloys, indicating that these stainless steel alloys would be safe to use in direct and prolonged contact with the skin. PMID- 14531868 TI - Allergic contact dermatitis from dicyclohexylmethane-4,4'-diisocyanate. AB - From August 1999 to April 2001, there was an outbreak of severe eczema at a factory manufacturing medical equipment. A glue, mainly based on the isocyanate dicyclohexylmethane-4,4'-diisocyanate (DMDI), was suspected as being the cause of the problem. 16 workers with recent episodes of eczema were patch tested with a standard series, an isocyanate series and work material. The latter consisted of, among other things, the glue, DMDI, and an amine, dicyclohexylmethane-4,4' diamine (DMDA), which is formed when DMDI reacts with water. 13 patients reacted to DMDI, 9 to 1,6-hexamethylenediisocyanate (HDI) and 4 to isophoronediisocyanate (IPDI), all of which are aliphatic isocyanates. None reacted to the aromatic isocyanates, diphenylmethane-4,4'-diisocyanate (MDI) or toluenediisocyanate (TDI). One explanation for this pattern could be that aromatic diisocyanates are more reactive than the aliphatic ones and that, therefore, they are inactivated before penetrating the skin. 5 patients reacted to DMDA and 5 to 4,4' diaminodiphenylmethane (MDA). Concurrent reactions to DMDA and or MDA with DMDI could be due to cross-reactivity. The positive reactions to MDA could also be a marker of MDI exposure. Yet another patient, investigated in 1997 with suspected work-related contact dermatitis from the glue, is described. She, however, showed no positive reactions to any isocyanates. PMID- 14531869 TI - Hydroxyisohexyl 3-cyclohexene carboxaldehyde- known as Lyral: quantitative aspects and risk assessment of an important fragrance allergen. AB - Hydroxyisohexyl 3-cyclohexene carboxaldehyde, also known as Lyral, is a fragrance ingredient identified as the cause of contact allergic reactions in 2-3% of eczema patients undergoing patch testing. Lyral has been included in the standard patch test series in many clinics due to its importance as an allergen. It has been used without restrictions in cosmetic products, until now. In the present study, the dose-response relationship of Lyral contact allergy was studied with doses relevant for normal exposure in cosmetic products. 18 eczema patients, who previously had given a positive patch test to Lyral 5% petrolatum, were included along with 7 control subjects. All cases were tested with a serial dilution of Lyral in ethanol 6% to 6 p.p.m and subjected to a 2-week, repeated open application test with a low dose of Lyral in ethanol. In the case of no reaction, this was followed by another 2 weeks of testing with a higher dose. The test was performed at the volar aspect of the forearm. In 16 of 18 cases (89%), a positive use test developed, 11 reacting to the low and 5 to the high concentration. None reacted to the vehicle control of ethanol applied to the contralateral arm. All controls were negative to both the test solutions of Lyral and the ethanol control. The difference between the test and the control group was statistically significant (Fisher's test, P < 0.001). It is concluded that Lyral at the current usage levels is inducing sensitization in the community. The same levels were shown to elicit allergic contact dermatitis in almost all sensitized individuals. A significant reduction in usage concentrations is recommended to prevent contact allergic reactions. PMID- 14531870 TI - Fragrance allergy in patients with hand eczema - a clinical study. AB - Fragrance allergy and hand eczema are both common among dermatological patients. Fragrance mix (FM) and its constituents have a recognized relevance to exposure to fine fragrances and cosmetic products. Based on extensive chemical analysis and database search, a new selection of fragrances was established, including 14 known fragrance allergens present in products to which hand exposure would occur. A non-irritating patch-test concentration for some fragrances was established in 212 consecutive patients. 658 consecutive patients presenting with hand eczema were patch tested with the European standard series and the developed selection of fragrances. 67 (10.2%) of the 658 patients had a positive reaction to 1 or more of our selection of fragrance chemicals present in the new selection. The most common reactions to fragrances not included in the FM were to citral, Lyral (hydroxyisohexyl-3-cyclohexene carboxaldehyde) and oxidized l-limonene. A concomitant reaction to the FM identified potential fragrance allergy in less than (1/2) of these patients. Exposure assessment and a statistically significant association between a positive patch test to our selected fragrances and patients' history support the relevance of this selection of fragrances. Those with a positive reaction to our selected fragrances were significantly more likely to have 1 or more positive patch tests in the standard series. This observation is the basis for the hypothesis concerning cross-reactivity and the effect of simultaneous exposure. The study found that fragrance allergy could be a common problem in patients with eczema on the hands. PMID- 14531872 TI - Oral lichen planus and dental materials: a case-control study. AB - The purpose of this study was to examine the association of dental materials with oral lichen planus (OLP) and, particularly, the effects of amalgam, amalgam corrosion status, gold and dissimilar metals in continuous contact. A case control study was performed with 43 OLP cases from the Oral Medicine Clinic at the School of Dentistry, University of Washington and 78 controls from a general dental care-screening clinic, also at the School of Dentistry, University of Washington. Health histories and oral examinations were obtained. Current metal or metal-based restorations were charted, along with corrosion status of amalgams and the presence of dissimilar metals in continuous contact. Adjusted odds ratios (ORs) and 95% confidence intervals for the following potential risk factors were found to be: (1) number of teeth with amalgam, OR = 1.02 (0.92, 1.13); (2) total surfaces of amalgam, OR = 0.96 (0.92, 1.0); (3) number of teeth with gold, OR = 1.12 (0.95, 1.31); (4) mean amalgam corrosion score, OR = 5.74 (2.34, 14.1); (5) presence of dissimilar metals in contact (per quadrant), OR = 1.25 (0.81, 1.92). These findings suggest that although the presence of amalgam or gold themselves is not associated with increased risk of OLP, corrosion of amalgams and the presence of a 'galvanic effect' from dissimilar dental materials in continuous contact (bimetallism) are associated with an increased risk of OLP. PMID- 14531871 TI - The fragrance hand immersion study - an experimental model simulating real-life exposure for allergic contact dermatitis on the hands. AB - Recently, we showed that 10 x 2% of consecutively patch-tested hand eczema patients had a positive patch test to a selection of fragrances containing fragrances relevant to hand exposure. In this study, we used repeated skin exposure to a patch test-positive fragrance allergen in patients previously diagnosed with hand eczema to explore whether immersion of fingers in a solution with or without the patch-test-positive fragrance allergen would cause or exacerbate hand eczema on the exposed finger. The study was double blinded and randomized. All participants had a positive patch test to either hydroxycitronellal or Lyral (hydroxyisohexyl 3-cyclohexene carboxaldehyde). Each participant immersed a finger from each hand, once a day, in a solution containing the fragrance allergen or placebo. During the first 2 weeks, the concentration of fragrance allergen in the solution was low (approximately 10 p.p.m.), whilst during the following 2 weeks, the concentration was relatively high (approximately 250 p.p.m.), imitating real-life exposure to a household product like dishwashing liquid diluted in water and the undiluted product, respectively. Evaluation was made using a clinical scale and laser Doppler flow meter. 3 of 15 hand eczema patients developed eczema on the finger immersed in the fragrance-containing solution, 3 of 15 on the placebo finger and 3 of 15 on both fingers. Using this experimental exposure model simulating real-life exposure, we found no association between immersion of a finger in a solution containing fragrance and development of clinically visible eczema on the finger in 15 participants previously diagnosed with hand eczema and with a positive patch test to the fragrance in question. PMID- 14531874 TI - Occupational allergic contact dermatitis due to thimerosal. PMID- 14531873 TI - Nummular allergic contact dermatitis after scabies treatment. PMID- 14531875 TI - Delayed sensitivity to Prolene. PMID- 14531876 TI - Allergic contact dermatitis from dodecyldiaminoethyl-glycine and isopropyl alcohol in a commercial disinfectant swab. PMID- 14531877 TI - Taxi driver's occupational allergic contact dermatitis from nickel in euro coins. PMID- 14531878 TI - Contact allergy to cystamine bis-lactamide. PMID- 14531879 TI - Allergic contact dermatitis due to 1,3-dichloropropene soil fumigant. PMID- 14531880 TI - Oral lichenoid reaction due to chromate/cobalt in dental prosthesis. PMID- 14531881 TI - Allergic contact dermatitis from cetearyl isononanoate. PMID- 14531882 TI - Occupational contact dermatitis from N-(2-(diethylamino)-ethyl) acrylamide. PMID- 14531883 TI - Allergic contact dermatitis from propylene glycol in Efudix cream. PMID- 14531884 TI - Occupational hand dermatitis in a wood turner due to rosewood (Dalbergia latifolia). PMID- 14531885 TI - Acrylic reactions: a review of 56 cases. PMID- 14531886 TI - Contact dermatitis from the antimicrobial coating of a desk mat. PMID- 14531887 TI - Maculopapular drug eruption due to the Japanese herbal medicine Kakkonto (kudzu or arrowroot decoction). PMID- 14531888 TI - Allergic contact dermatitis from medium-chain triglycerides. PMID- 14531890 TI - Invasion and survival strategies of Anaplasma phagocytophilum. AB - Anaplasma phagocytophilum is an aetiological agent of human granulocytic ehrlichiosis, an emerging tick-borne zoonosis in the United States and Europe. This obligate intracellular bacterium is unique in that it colonizes polymorphonuclear leucocytes (neutrophils). Neutrophils are key players in innate immunity. These short-lived phagocytes ingest invading microorganisms and destroy them by various means, which include fusing the bacteria-containing phagosome with acidic lysosomes as well as directing toxic oxidative and proteolytic compounds into the phagosomal lumen. Its tropism for neutrophils indicates that A. phagocytophilum uses strategies for evading and/or neutralizing these microbicidal activities. This review focuses on some of the mechanisms that A. phagocytophilum uses for neutrophil adhesion, surviving within the hostile intracellular environment of its host neutrophil and for effectively disseminating to naive host cells. PMID- 14531891 TI - Innate immunity and biodefence vaccines. AB - Host defence in vertebrates is achieved by the integration of two distinct arms of the immune system: the innate and adaptive responses. The innate response acts early after infection (within minutes), detecting and responding to broad cues from invading pathogens. The adaptive response takes time (days to weeks) to become effective, but provides the fine antigenic specificity required for complete elimination of the pathogen and the generation of immunologic memory. Antigen-independent recognition of pathogens by the innate immune system leads to the rapid mobilization of immune effector and regulatory mechanisms that provide the host with three critical advantages: (i) initiating the immune response (both innate and adaptive) and providing the inflammatory and co-stimulatory context for antigen recognition; (ii) mounting a first line of defence, thereby holding the pathogen in check during the maturation of the adaptive response; and (iii) steering the adaptive immune system towards the cellular or humoral responses most effective against the particular infectious agent. The quest for safer and more effective vaccines and immune-based therapies has taken on a sudden urgency with the increased threat of bioterrorism. Only a handful of vaccines covering a small proportion of potential biowarfare agents are available for human use (e.g. anthrax and small pox) and these suffer from poor safety profiles. Therefore, next generation biodefence-related vaccines and therapies with improved safety and the capacity to induce more rapid, more potent and broader protection are needed. To this end, strategies to target both the innate and adaptive immune systems will be required. PMID- 14531892 TI - Lethal toxin from Clostridium sordellii induces apoptotic cell death by disruption of mitochondrial homeostasis in HL-60 cells. AB - Lethal toxin (LT) from Clostridium sordellii (strain IP82) inactivates in glucosylating the small GTPases Ras, Rap, Ral and Rac. In the present study we show that LT-IP82 induces cell death via an intrinsic apoptotic pathway in the myeloid cell-line HL-60. LT-IP82 was found to disrupt mitochondrial homeostasis as characterized by a decrease in mitochondrial transmembrane potential and cardiolipin alterations, associated with the release of cytochrome c in the cytosol. Time-course studies of caspase activation revealed that caspase-9 and caspase-3 were activated before caspase-8. Moreover, although LT-IP82-induced cell death was abrogated by caspase-inhibitors, these inhibitors did not suppress mitochondrial alterations, indicating that caspase activation occurs downstream of mitochondria. Protection of mitochondria by Bcl-2 overexpression prevented mitochondrial changes as well as apoptosis induction. Furthermore, evidence is provided that LT-IP82-induced apoptosis is not a consequence of cortical actin disorganization, suggesting that Rac inactivation does not initiate the apoptotic process. Cell exposure to LT-IP82 leads to a co-localization of the toxin with a mitochondrial marker within 2 h. Therefore, we suggest that LT-IP82 could act at the mitochondrion level independently of its enzymatic effect on small GTPases. PMID- 14531893 TI - Synergistic roles for the Map and Tir effector molecules in mediating uptake of enteropathogenic Escherichia coli (EPEC) into non-phagocytic cells. AB - Enteropathogenic Escherichia coli (EPEC) are a major cause of paediatric diarrhoea and a model for the family of attaching and effacing (A/E) pathogens. Enteropathogenic Escherichia coli encode a type III secretion system (TTSS) to transfer effector proteins into host cells, a process which is essential for virulence. In addition to generation of A/E lesions, the TTSS is also implicated in the ability of EPEC to invade cultured cells but the effector proteins responsible for promoting invasion have not been identified. In this paper we confirm the requirement of TTSS in EPEC invasion and demonstrate important roles for the Map and Tir effector molecules. Whereas in trans expression of Tir in the tir mutant restored invasion to wild-type levels, similar complementation of the map mutation by in trans expression of Map results in a hyperinvasive phenotype. The Map effector protein has two distinct functions within host cells, mediating Cdc42-dependent filopodia formation and targeting mitochondria to elicit dysfunction. The former function appears to be related to Map's ability to promote invasion as this was inhibited by interference with Cdc42 signalling. Conversely, Map targeting to mitochondria is not necessary for invasion. Promotion of EPEC invasion by Tir appears to involve interaction with intimin but is independent of pedestal formation, and intimin-Tir interaction is neither necessary nor sufficient for invasion. Comparison of the invasiveness of strains lacking Tir and/or Map with wild-type or mutant strains expressing the effectors in trans provides evidence that Map and Tir stimulate invasion by synergistic mechanisms. This synergism, which is in stark contrast to the antagonistic actions of Map and Tir in regulating filopodia and pedestal formation, further illustrates the complex interplay between EPEC effectors. PMID- 14531894 TI - More than just innate immunity: comparative analysis of Chlamydophila pneumoniae and Chlamydia trachomatis effects on host-cell gene regulation. AB - Chlamydophila pneumoniae and Chlamydia trachomatis cause infections of the respiratory or urogenital tract. In addition, both species have been associated with atherosclerosis or reactive arthritis respectively. For these intracellular pathogens the interaction with their host-cells is of particular importance. To get insight into this relationship, we conducted a comparative analysis of the host-cell gene regulation of human epithelial cells during infection with Chlamydia. In a screening of HeLa cells by Affymetrix-microchips, numerous regulated host-genes were identified. A detailed expression profile was obtained for 14 genes by real-time RT-PCR - comparing C. pneumoniae, C. trachomatis and intracellular S. typhimurium. The transcriptional responses induced by C. pneumoniae were similar (but usually smaller) compared to C. trachomatis, some were absent. UV-inactivated bacteria induced no differential gene expression suggesting that pathomechanisms other than those associated with innate immunity play here an important role. The expression pattern induced by Salmonella differed substantially. These genus- or group-specific transcriptional response patterns elicited by viable intracellular pathogens may considerably contribute to the different pathologies encountered in the clinic. PMID- 14531895 TI - The ShiA protein encoded by the Shigella flexneri SHI-2 pathogenicity island attenuates inflammation. AB - Shigella spp. are the aetiologic agents of dysentery, a severe diarrhoeal syndrome characterized by acute inflammation in the colon. The inflammatory response, which includes recruitment of polymorphonuclear leukocytes (PMN), damages the colonic mucosa and exacerbates the infection. Shigella encodes a pathogenicity island (PAI), SHI-2, which is localized in a region of the chromosome linked to the induction of inflammation. Surprisingly, SHI-2 deletion mutants induce a stronger inflammatory response than wild-type Shigella as measured by increased villus blunting, increased PMN infiltration and induction of apoptosis in a rabbit ileal loop model of shigellosis. Mutational analysis mapped the hyper-inflammatory phenotype to a single gene, shiA. Similar to SHI-2 deletion mutants, infection with a shiA mutant strain induces dramatically elevated levels of inflammation when compared to the wild-type strain. Furthermore, infection with a wild-type strain containing multiple copies of shiA results in fewer infiltrating PMN and apoptotic cells, as well as preservation of a normal villus architecture at the site of infection, thus acting in a dominant fashion over the pro-inflammatory mechanisms of Shigella. The molecular mechanism of action of ShiA is independent of any in vitro phenotype associated with Shigella virulence. Our data suggest that ShiA allows Shigella to attenuate the host inflammatory response in a novel manner. PMID- 14531896 TI - Obligatory intracellular parasitism by Ehrlichia chaffeensis and Anaplasma phagocytophilum involves caveolae and glycosylphosphatidylinositol-anchored proteins. AB - Obligatory intracellular, human ehrlichiosis agents Ehrlichia chaffeensis and Anaplasma phagocytophilum create unique replicative compartments devoid of lysosomal markers in monocytes/macrophages and granulocytes respectively. The entry of these bacteria requires host phospholipase C (PLC)-gamma2 and protein tyrosine kinases, but their entry route is still unclear. Here, using specific inhibitors, double immunofluorescence labelling and the fractionation of lipid rafts, we demonstrate that bacterial entry and intracellular infection involve cholesterol-rich lipid rafts or caveolae and glycosylphosphatidylinositol (GPI) anchored proteins. By fluorescence microscopy, caveolar marker protein caveolin-1 was co-localized with both early and replicative bacterial inclusions. Additionally, tyrosine-phosphorylated proteins and PLC-gamma2 were found in bacterial early inclusions. In contrast, clathrin was not found in any inclusions from either bacterium. An early endosomal marker, transferrin receptor, was not present in the early inclusions of E. chaffeensis, but was found in replicative inclusions of E. chaffeensis. Furthermore, several bacterial proteins from E. chaffeensis and A. phagocytophilum were co-fractionated with Triton X-100 insoluble raft fractions. The formation of bacteria-encapsulating caveolae, which assemble and retain signalling molecules essential for bacterial entry and interact with the recycling endosome pathway, may ensure the survival of these obligatory intracellular bacteria in primary host defensive cells. PMID- 14531897 TI - Helicobacter pylori stimulates host cyclooxygenase-2 gene transcription: critical importance of MEK/ERK-dependent activation of USF1/-2 and CREB transcription factors. AB - Cyclooxygenase-2 (COX-2) represents the inducible key enzyme of arachidonic acid metabolism and contributes to the pathogenesis of gastroduodenal ulcers and gastric cancer. Helicobacter pylori infection is associated with elevated gastric COX-2 levels, but the mechanisms underlying H. pylori-dependent cox-2 gene expression are unclear. H. pylori stimulated cox-2 mRNA and protein abundance in gastric epithelial cells in vitro and in vivo, and functional analysis of the cox 2 gene promoter mapped its H. pylori-responsive region to a proximal CRE/Ebox element at -56 to -48. Moreover, USF1/-2 and CREB transcription factors binding to this site were identified to transmit H. pylori-dependent cox-2 transcription. Activation of MEK/ERK1/-2 signalling by bacterial virulence factors located outside the H. pylori cag pathogenicity island (cagPAI) was found to mediate bacterial effects on the cox-2 promoter. Our study provides a detailed description of the molecular pathways underlying H. pylori-dependent cox-2 gene expression in gastric epithelial cells, and may thus contribute to a better understanding of mechanisms underlying H. pylori pathogenicity. PMID- 14531898 TI - Invasion of human epithelial cells by Campylobacter upsaliensis. AB - Few data exist on the interaction of Campylobacter upsaliensis with host cells, and the potential for this emerging enteropathogen to invade epithelial cells has not been explored. We have characterized the ability of C. upsaliensis to invade both cultured epithelial cell lines and primary human small intestinal cells. Epithelial cell lines of intestinal origin appeared to be more susceptible to invasion than non-intestinal-derived cells. Of three bacterial isolates studied, a human clinical isolate, CU1887, entered cells most efficiently. Although there was a trend towards more efficient invasion of Caco-2 cells by C. upsaliensis CU1887 at lower initial inocula, actual numbers of intracellular organisms increased with increasing multiplicity of infection and with prolonged incubation period. Confocal microscopy revealed C. upsaliensis within primary human small intestinal cells. Both Caco-2 and primary cells in non-confluent areas of the infected monolayers were substantially more susceptible to infection than confluent cells. The specific cytoskeletal inhibitors cytochalasin B, cytochalasin D and vinblastine attenuated invasion of Caco-2 cells in a concentration-dependent manner, providing evidence for both microtubule- and microfilament-dependent uptake of C. upsaliensis. Electron microscopy revealed the presence of organisms within Caco-2 cell cytoplasmic vacuoles. C. upsaliensis is capable of invading epithelial cells and appears to interact with host cell cytoskeletal structures in order to gain entry to the intracellular environment. Entry into cultured primary intestinal cells ex vivo provides strong support for the role of host cell invasion during human enteric C. upsaliensis infection. PMID- 14531899 TI - Malignant sarcoma masquerading as a pseudotumour in a patient with mild haemophilia. PMID- 14531900 TI - Atypical large granular lymphocytes in a child. PMID- 14531901 TI - The role of haploidentical stem cell transplantation in the management of children with haematological disorders. AB - The broader application of stem cell transplantation (SCT) for paediatric diseases has been limited by a lack of human leucocyte antigen (HLA)-matched donors. Virtually all children, however have at least one haploidentical parent who could serve as a donor. Such a donor is immediately available and the considerable costs of additional HLA typing, registry and banking expenditures that are necessary to procure an unrelated donor, could be reduced. Recent technological advances appear to have overcome the historical problems of graft rejection and severe graft versus host disease in the haploidentical setting, and in the latest studies the overall survival for children undergoing haploidentical SCT for leukaemia is now comparable with that following unrelated donor bone marrow or cord blood transplantation. Post-transplant infectious complications and leukaemia relapse remain the most important barriers yet to overcome, and new directions in the use of adoptive cellular immunity appear to be promising in this respect. Haploidentical SCT is now a viable option for those children who do not have an HLA compatible sibling or fully matched unrelated donor. The relative merits of a haploidentical family donor versus mismatched unrelated bone marrow or cord blood donation needs to be assessed in prospective, randomized clinical trials. PMID- 14531902 TI - Haematology at the Hammersmith Hospital and Royal Postgraduate Medical School 1934-1994. PMID- 14531903 TI - Cytogenetic findings in adult de novo acute myeloid leukaemia. A population-based study of 303/337 patients. AB - During a 10-year period (1992-2001) in the region of Southern Denmark, 337 patients aged 15 years or older (range 16-93 years, median 67 years) were diagnosed with acute myeloid leukaemia (AML). Cytogenetic analysis was carried out in 90%, of whom 53% had clonal chromosome aberrations. Some 24% and 31% had only numerical or structural abnormalities respectively. The remaining patients showed both types of abnormalities. Ploidy levels in decreasing order were: pseudodiploidy, 41%; hyperdiploidy, 32%; and hypodiploidy, 27%. Pseudodiploidy characterizes type M3 (70%) and hypodiploidy M6 (56%). Recurrent cytogenetic abnormalities--t(8;21), t(15;17) and inv(16)--were found in 3.3%, 3.3% and 2.0% of all patients respectively. Prognostically intermediate and adverse aberrations were found in 39% and 44%, respectively, of those with an abnormal karyotype. Rare recurrent aberrations were found in two patients in this material. A previously described non-recurrent abnormality was found to be recurrent in one patient [der(20)t(11;20)(q13.2;p13)]. New, previously undescribed abnormalities were found in 41 patients. Statistically significant correlations were found between t(15;17) and young age (P < 0.001), inv(16) and young age (P < 0.006), 17 and M6 (P = 0.007), and M6 and complex karyotype with five or more unrelated aberrations (P = 0.004). We conclude that this truly population-based cytogenetic study of adult AML showed distributions of chromosome abnormalities that differ from those described so far. PMID- 14531904 TI - Possible regulation of Wilms' tumour gene 1 (WT1) expression by the paired box genes PAX2 and PAX8 and by the haematopoietic transcription factor GATA-1 in human acute myeloid leukaemias. AB - Overexpression of the embryonic transcription factor, Wilms' tumour protein 1 (WT1), is common in acute myeloid leukaemias (AML). Mutations of Wilms' tumour gene 1 (WT1) in AML are rare and WT1 expression may be increased by other transcription factors. PAX2, PAX8 and GATA-1 are known physiological regulators of WT1. In the present study, we analysed either bone marrow or blood samples of 43 AML patients for the expression levels of WT1, PAX2, PAX8 and GATA-1 by real time reverse transcription polymerase chain reaction (LightCycler). Bone marrow samples of patients without haematological malignancies and stem cell preparation samples from healthy donors and lymphoma patients served as controls. PAX2 expression was found in 11 of 43 AML samples, with a clear correlation of PAX2 with WT1 expression levels observed. PAX8 expression was found in two additional samples. GATA-1 expression was detectable in 41 of 43 AML samples and also in all control samples; no significant differences between these groups were observed and no correlation of GATA-1 expression with WT1 expression levels was apparent. In conclusion, PAX2, and possibly PAX8, appears to be a candidate for the upregulation of WT1 in a proportion of AML, whereas GATA-1 expression cannot be explained as an inducer of WT1. In two-thirds of leukaemias from our series, the basis of WT1 upregulation cannot be explained by the simple upregulation of the known WT1 activators. PMID- 14531905 TI - Clinical significance of residual disease during treatment in childhood acute myeloid leukaemia. AB - In children with acute myeloid leukaemia (AML), morphological and karyotypic studies cannot precisely assess response to treatment, and less than one-third of patients have genetic markers for molecular studies of residual disease. We determined the usefulness of a four-colour flow cytometric strategy developed in our laboratory to study residual disease. We first compared the immunophenotypes of AML cells obtained from 54 children at diagnosis with those of cells from 59 normal or regenerating bone marrow samples. Forty-six of the 54 AML cases (85.2%) had immunophenotypes that allowed detection of 0.1-0.01% residual leukaemic cells. Of 230 bone marrow samples obtained from those 46 patients during and off treatment, 61 (26.5%) had >/= 0.1% AML cells by flow cytometry. We found that core binding factor-associated AML had a significantly better early treatment response. Mean (+/- standard error) 2-year survival estimate was 33.1 +/- 19.1% for patients with >/= 0.1% AML cells by flow cytometry after induction therapy, but 72.1 +/- 11.5% for those with < 0.1% AML cells (P = 0.022); overt recurrence of AML within the subsequent 6 months was significantly more likely in the former group. The assay described here holds promise for guiding the choice of post remission treatment options in children with AML. PMID- 14531906 TI - Occurrence of dysregulated oncogenes in primary plasma cells representing consecutive stages of myeloma pathogenesis: indications for different disease entities. AB - This study investigated the expression pattern in primary plasma cells (PCs) of putative oncogenes suggested to be involved in multiple myeloma (MM) development. cDNA archives were generated by global reverse transcription polymerase chain reaction from CD38++/CD19-/CD56-/++ aberrant PCs of a prospective cohort of 96 subjects, including healthy individuals, patients with monoclonal gammopathies of undetermined significance (MGUS), MM and MM with extramedullary manifestations (ExMM). The cDNA archives were analysed quantitatively for expression of the cyclin D1, fibroblast growth factor receptor 3 (FGFR3), C-MYC, C-MAF and cyclin D3 oncogenes. In addition, all patients were screened for IGH-MMSET hybrid transcripts. None of the analysed oncogenes was randomly distributed. C-MYC and cyclin D3 expression increased at the extramedullary transformation stage. Furthermore, C-MYC and cyclin D3 expression in CD56+ MM was similar to MGUS, whereas CD56- MM was similar to ExMM. FGFR3/IGH-MMSET was only observed among CD56+ MM patients, whereas an increased frequency of C-MAF dysregulation was seen among CD56- MM. High cyclin D1 expression levels were identified at similar frequencies at all stages, whereas the frequency of patients with low cyclin D1 levels increased during MM development. These data support the stepwise transformation model accumulating genetic alterations and proliferative capacity during MM initiation and development resulting in different clinical entities. PMID- 14531907 TI - Enhanced production of osteopontin in multiple myeloma: clinical and pathogenic implications. AB - In this study, we examined osteopontin (OPN) production in myeloma cells and plasma OPN levels in multiple myeloma (MM) patients. We assessed OPN production in bone marrow cells (BMCs) by immunocytochemistry and enzyme-linked immunosorbent assay (ELISA). We also assessed OPN production in various B-cell malignant cell lines, including three myeloma cell lines by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. In addition, we measured plasma OPN concentrations by ELISA in 30 MM patients, 21 monoclonal gammopathy of undetermined significance (MGUS) patients and 30 healthy volunteers. As a result, in an immunocytochemical study, abundant OPN was detected in BMCs from overt MM patients, whereas no OPN was detected in BMCs from patients with other haematological diseases, including MGUS. Cultured BMCs from overt MM patients produced more OPN than those from patients with either smouldering MM or MGUS. Myeloma cell lines spontaneously produced OPN. Plasma OPN levels of MM patients were significantly higher than those of MGUS patients and healthy volunteers (P < 0.05). Moreover, they correlated with both progression and bone destruction of the disease (P < 0.05). These suggest that myeloma cells actively produce OPN, which possibly contributes to osteoclastic bone resorption in MM. Plasma OPN levels may be a useful biomarker for assessing bone destruction in MM and distinguishing MM from MGUS or smouldering MM. PMID- 14531908 TI - Chlorambucil in combination with induction and maintenance rituximab is feasible and active in indolent non-Hodgkin's lymphoma. AB - We investigated the toxicity and efficacy of the chimaeric anti-CD20 antibody rituximab in combination with standard-dose chlorambucil in newly diagnosed and relapsed/refractory indolent B-cell lymphoma patients. A total of 29 patients (15 newly diagnosed and 14 relapsed/refractory) with low-grade or follicular B-cell non-Hodgkin's lymphoma (NHL) were included in this phase II study. Therapy consisted of chlorambucil 6 mg/m2/d for 6 consecutive weeks in combination with a standard 4-weekly rituximab administration schedule in the induction phase. Patients responding to the induction therapy received four additional cycles with chlorambucil (6 mg/m2/d for 2 weeks/month) plus rituximab (once a month). Twenty six patients (89%) completed the treatment; only one patient discontinued treatment because of haematological toxicity. At the end of the study, the dose of chlorambucil had to be reduced in seven patients (27%) and six patients (23%) required a delay in further treatment, as a result of toxicity during consolidation therapy. Only one patient was withdrawn from the study because of progressive disease; the 27 patients evaluable for response at the end of consolidation achieved a clinical response (63% complete response and 26% partial response). A significant CD4+ and CD56+ depletion was observed after induction and during consolidation therapy; two herpes zoster virus infections and one perianal abscess represented major infectious morbidities registered during the study. Based on our preliminary data, the combination of chlorambucil with rituximab seemed to be well tolerated and active. Its definitive role in the treatment of low-grade NHL should be further evaluated in randomized trials. PMID- 14531909 TI - Successful treatment of refractory pure red cell aplasia associated with lymphoproliferative disorders with the anti-CD52 monoclonal antibody alemtuzumab (Campath-1H). AB - Acquired pure red cell aplasia (PRCA) is a rare, but significant, complication of lymphoproliferative disorders. It is characterized by anaemia, absence of red cell precursors in the bone marrow and normal granulopoiesis and megakaryopoiesis. We describe two patients with refractory pure red cell aplasia associated with chronic lymphocytic leukaemia (CLL) and a large granular CD8 T lymphocytic leukaemia (LGL) respectively. Both patients had failed multiple treatment regimens for PRCA and were transfusion dependent. Both patients were subsequently treated with the anti-CD52 humanized monoclonal antibody, alemtuzumab, receiving total doses in excess of 300 mg. Response to treatment, as documented by a rapid increase in the reticulocyte count, occurred as early as the third infusion. At the time of this report, both patients remain in complete remission with normal haemoglobin levels. Alemtuzumab appears to be an effective and well-tolerated therapy for pure red blood cell aplasia associated with lymphoproliferative disorders. PMID- 14531910 TI - Predictive value of early 18F-fluoro-deoxyglucose positron emission tomography in chemosensitive relapsed lymphoma. AB - 18F-fluoro-deoxyglucose (FDG) positron emission tomography (PET) might be a better tool than computerized tomography (CT) in predicting long-term treatment outcome in patients with relapsed chemosensitive lymphoma who are candidates for autologous stem cell transplantation (ASCT). We studied patients with recurrent or persistent aggressive non-Hodgkin's lymphoma (NHL) and Hodgkin's disease (HD), who were treated with three courses of second-line induction chemotherapy [DHAP VIM (dexamethasone, cytarabine, cisplatin followed by etoposide, iphosphamide and methotrexate)-DHAP], followed by myeloablative therapy and ASCT if chemosensitive. FDG-PET was performed in parallel to conventional diagnostic methods before starting, and after two courses of, second-line therapy. Of 68 relapsed lymphoma patients, 46 chemosensitive patients (33 NHL and 13 HD) were included, of whom 39 were transplanted. After DHAP-VIM, the second PET scan was normalized in 15/46 patients; progression-free survival at 2 years was 62% for PET-negative patients versus 32% for PET-positive patients (P = 0.048). The relative risk for progressive disease in patients with < 90% intensity reduction was 2.85 (95% confidence interval 1.15-7.05, P = 0.018). Early FDG-PET may help to predict the long-term treatment outcome of ASCT in chemosensitive patients with relapsed lymphoma and identify those patients who need extra or alternative treatment. Disappearance or > 90% reduction of intensity of abnormal FDG uptake after two courses of reinduction therapy was correlated with a favourable outcome. PMID- 14531911 TI - DNA microarray analysis of stage progression mechanism in myelodysplastic syndrome. AB - Myelodysplastic syndrome (MDS) is a clonal disorder of haematopoietic stem cells. Despite the high incidence of MDS in the elderly, effective treatment of individuals in its advanced stages is problematic. DNA microarray analysis is a potentially informative approach to the development of new treatments for MDS. However, a simple comparison of 'transcriptomes' of bone marrow mononuclear cells among individuals at distinct stages of MDS would result in the identification of genes whose expression differences only reflect differences in the proportion of MDS blasts within bone marrow. Such a 'population shift' effect has now been avoided by purification of haematopoietic stem-like cells that are positive for the cell surface marker AC133 from the bone marrow of healthy volunteers and 30 patients at various stages of MDS. Microarray analysis with the AC133+ cells from these individuals resulted in the identification of sets of genes with expression that was specific to either indolent or advanced stages of MDS. The former group of genes included that for PIASy, which catalyses protein modification with the ubiquitin-like molecule SUMO. Induction of PIASy expression in a mouse myeloid cell line induced apoptosis. A loss of PIASy expression may therefore contribute directly to the growth of MDS blasts and stage progression. PMID- 14531912 TI - Granulomatous slack skin disease--disease features and response to pentostatin. AB - Granulomatous slack skin disease (GSSD) is a rare condition characterized clinically by redundant skin folds, which show a predilection towards flexural areas, and histologically by a granulomatous T-cell infiltrate and loss of elastic fibres. The disease is often indolent, although rapid progression and transformation have been described. There is much debate as to whether this condition is a subset of mycosis fungoides or a separate disease entity in itself. We describe a case of GSSD with unique manifestations including granulomatous bone marrow involvement and hypercalcaemia. The patient has twice achieved a good response to pentostatin after failure of combination chemotherapy. This is the first report of the successful use of the purine analogue pentostatin in the management of GSSD. PMID- 14531913 TI - Plasma levels of tumour necrosis factor alpha and interleukin-6 predict progression-free survival following thalidomide therapy in patients with previously untreated multiple myeloma. AB - We studied marrow angiogenesis and plasma levels of angiogenic cytokines in 38 patients receiving thalidomide therapy for previously untreated myeloma. The effect of therapy and the relationship of cytokine levels to myeloma cell proliferation, bone marrow microvessel density and progression-free survival (PFS) were studied. High pretreatment tumour necrosis factor-alpha (TNFalpha) levels (> 11 pg/ml) and increased interleukin (IL)-6 of > 2 pg/ml predicted for poorer PFS (TNFalpha, 48% versus 74% at 2 years, P = 0.01; IL-6, 24% versus 70% at 2 years, P = 0.01). None of the other parameters predicted response or PFS, and no significant changes in cytokine levels occurred with therapy. PMID- 14531914 TI - T- and B-cell immune reconstitution and clinical outcome in patients with multiple myeloma receiving T-cell-depleted, reduced-intensity allogeneic stem cell transplantation with an alemtuzumab-containing conditioning regimen followed by escalated donor lymphocyte infusions. AB - Immune reconstitution after conventional allogeneic transplantation is a major determinant of survival. We conducted a detailed investigation of T- and B-cell immune reconstitution and clinical outcome in 19 patients with multiple myeloma undergoing reduced-intensity stem cell transplantation using in vivo T-cell depletion with alemtuzumab. These patients experienced delayed T-cell recovery, particularly in the naive (CD45 RA+) CD4 compartment. T-cell receptor spectratype analysis showed a reduced repertoire diversity, which improved rapidly after the administration of donor leucocyte infusions and subsequent conversion to full donor T-cell chimaerism. Post-transplant recovery of CD19+ B cells was also delayed for up to 18 months. Spectratype analysis of IgH CDR3 repertoire revealed a gradual normalization in IgM spectratype complexity by 6-12 months after transplant. There was a high incidence of viral infection, particularly cytomegalovirus reactivation, but the regimen-related mortality was low, perhaps because of the very low incidence of acute graft-versus-host disease (GVHD; grade I-II skin GVHD was seen in 5/19 patients). Over 80% of all patients have relapsed at a median of 283 (range 153-895) d after transplant, suggesting that the initially low rate of GVHD comes at a high price with regard to the desired graft versus-myeloma effect. PMID- 14531915 TI - T-cell alloreactivity dominates natural killer cell alloreactivity in minimally T cell-depleted HLA-non-identical paediatric bone marrow transplantation. AB - Natural killer (NK) cell alloreactivity resulting from killer immunoglobulin-like receptor (KIR) ligand incompatibility improves outcomes in patients receiving extensively T-cell-depleted bone marrow (BM) grafts. Patients with KIR ligand incompatibility are at risk for donor T-cell alloreactivity. We investigated the relative significance of NK-cell and T-cell alloreactivity in 105 paediatric patients who received a minimally T-cell-depleted human leucocyte antigen-non identical BM transplantation. Donor NK-cell incompatibility did not improve patient outcome [engraftment, graft-versus-host disease (GVHD), relapse or overall survival]. In contrast, donor T-cell incompatibility was a risk factor for acute GVHD, chronic GVHD and death. Thus, T-cell alloreactivity dominated that of NK cells in minimally T-cell-depleted grafts. PMID- 14531916 TI - Pregnancy-associated venous thromboembolism (VTE) in combined heterozygous factor V Leiden (FVL) and prothrombin (FII) 20210 A mutation and in heterozygous FII single gene mutation alone. AB - The risk of venous thromboembolism (VTE) in the absence of prophylaxis was evaluated in a retrospective study of 47 women (84 pregnancies) with combined thrombophilia [heterozygous factor V Leiden (FVL) plus prothrombin (FII) 20210A mutation (group I)] and in 82 women (193 pregnancies) with the FII alone (group II). VTE was more frequent in group I than in group II [17.8% versus 6.2%, P = 0.003, relative risk (RR) 2.9, 95% confidence interval (CI) 1.4-5.9], ante partum (7.1% and 2.1%) and post partum (11.5% and 4.2%). The risk was higher in index cases than in family members (RR 2.5, 95% CI 1.2-5.2 and RR 2.1, 95% CI 0.2-22.3 respectively) Even women who had no history of VTE before pregnancy had an increased risk (RR 2.2, 95% CI 1.0-4.8). Our results suggest that, during ante partum, prophylaxis is indicated in women with combined thrombophilia and with a VTE before pregnancy. In those without VTE before pregnancy, prophylaxis might be decided for each individual case, taking into consideration all risk factors. In women with the FII mutation alone, the low risk may not justify prophylaxis in the absence of previous VTE. In post partum, prophylaxis is indicated in all cases. PMID- 14531917 TI - A multicentre assessment of the endogenous thrombin potential using a continuous monitoring amidolytic technique. AB - This study assessed the inter-laboratory imprecision associated with the measurement of the endogenous thrombin potential (ETP). The initial studies used techniques that had evolved in each of the participating laboratories. Samples from normal healthy subjects (n = 10), two patients receiving coumarin therapy [International Normalized Ratio approximately 2.0 and approximately 4.0] and a further two subjects receiving treatment with unfractionated heparin (anti-Xa 0.07 i.u./ml and 0.31 i.u./ml) were assayed relative to a lyophilized normal plasma that had arbitrarily been assigned a potency of 100%. Considerable variation in potency estimates was observed between the centres, although individual laboratories using fully automated techniques achieved acceptable levels of imprecision as assessed by the coefficient of variation (CV) (intra assay CV < 9.5%, inter-assay CV < 12.5%). A second study to assess a similar range of samples, using a standardized assay protocol and incorporating appraisal of two chromogenic substrates, CBS.0068 or Pefachrom TG, demonstrated markedly improved agreement in potency estimates between centres and good correlation (r > 0.96) between the chromogenic substrates. Our data demonstrates that an automated ETP method can be standardized between laboratories and suitable levels of imprecision achieved, using different analysers (COBAS Mira at two centres and an ACL-300R) and two thrombin substrates. This indicates that more widespread use of ETP measurements in clinical laboratories is feasible. PMID- 14531918 TI - Two novel factor V null mutations associated with activated protein C resistance phenotype/genotype discrepancy. AB - Activated protein C (APC) resistance phenotype/genotype discrepancy is a very rare event. The objective of this study was to characterize the molecular mechanisms in two cases of APC phenotype/genotype discrepancy. An approach using direct sequencing of each exon and splicing junctions of the factor V gene showed that two novel factor V null mutations combined with heterozygous factor V Leiden mutation were responsible for this discrepancy. Our results suggest the necessity to use both phenotypic and genotypic analyses in some cases to determine an accurate diagnosis. PMID- 14531919 TI - Familial polycythemia vera with Budd-Chiari syndrome in childhood. AB - Polycythemia vera is a myeloproliferative disorder that, in most cases, occurs sporadically with a median age at presentation of 60 years. Familial cases are very rare and usually manifest in elderly family members. The Budd-Chiari syndrome, characterized by the obstruction and occlusion of the suprahepatic veins, is a rare typical complication in polycythemia vera patients. To date, only two children or adolescents with polycythemia vera and Budd-Chiari syndrome have been described. Here, we report an 11-year-old girl with Budd-Chiari syndrome as the initial symptom of familial polycythemia vera, which was also found in the girl's grandmother. Details of the diagnostic procedures used and the clinical course are reported. The patient underwent orthotopic liver transplantation and is being treated with hydroxyurea. The available literature on familial polycythemia vera and polycythemia vera in childhood with and without Budd-Chiari syndrome is reviewed. PMID- 14531920 TI - Heterocellular hereditary persistence of fetal haemoglobin affects the haematological parameters of beta-thalassaemia trait. AB - To assess and define the effects of heterocellular hereditary persistence of fetal haemoglobin (HPFH) on the haematological phenotype of heterozygous beta thalassaemia, we have studied a large kindred that included a total of 204 subjects with 60 beta-thalassaemia carriers, of whom 35 were also heterozygous, and five homozygous, for heterocellular HPFH. The study was possible because of the homogeneity of the beta-thalassaemia mutation and the ability to genotype the heterocellular HPFH allele. Heterocellular HPFH had a significant effect on the mean corpuscular haemoglobin (MCH), mean corpuscular volume (MCV) and haemoglobin (Hb) A2 values in the beta-thalassaemia carriers and accounted for 29%, 30% and 24% of their respective variances. beta-thalassaemia subjects with heterocellular HPFH had higher MCV and MCH values, concomitant with lower levels of Hb A2, and a reduced ineffective erythropoiesis. We conclude that co-inheritance of heterocellular HPFH leads to a primary increase in gamma-chain synthesis in beta thalassaemia trait and can be another confounding factor in the use of red cell indices and Hb A2 levels in population screening for beta-thalassaemia. PMID- 14531921 TI - Causes of death in sickle cell disease: an autopsy study. AB - More precise analysis of causes of death is needed to focus research efforts and improve morbidity and mortality in sickle cell disease. In this study, the morphological evidence of the cause of death was studied in 306 autopsies of sickle cell disease, which were accrued between 1929 and 1996. The most common cause of death for all sickle variants and for all age groups was infection (33 48%). The terminal infection was heralded by upper respiratory tract syndromes in 72.6% and by gastroenteritis in 13.7%. The most frequent portal of entry in children was the respiratory tract but, in adults, a site of severe chronic organ injury. Other causes of death included stroke 9.8%, therapy complications 7.0%, splenic sequestration 6.6%, pulmonary emboli/thrombi 4.9%, renal failure 4.1%, pulmonary hypertension 2.9%, hepatic failure 0.8%, massive haemolysis/red cell aplasia 0.4% and left ventricular failure 0.4%. Death was frequently sudden and unexpected (40.8%) or occurred within 24 h after presentation (28.4%), and was usually associated with acute events (63.3%). This study shows that the first 24 h after presentation for medical care is an especially perilous time for patients with sickle cell disease and an acute event. Close monitoring and prompt aggressive treatment are warranted. PMID- 14531922 TI - Concomitant myelodysplastic syndrome and chronic myeloid leukaemia: treatment outcomes with imatinib mesylate. PMID- 14531923 TI - Severe acute respiratory syndrome and lupus anticoagulants in children. PMID- 14531924 TI - Malignancies in sickle cell disease patients treated with hydroxyurea. PMID- 14531925 TI - Lithium effects on neutrophil motility in Shwachman-Diamond syndrome: evaluation by computer-assisted image analysis. PMID- 14531926 TI - Prognostic features of splenic lymphoma with villous lymphocytes. PMID- 14531927 TI - Erythrocytosis and the Chuvash von Hippel-Lindau mutation. PMID- 14531928 TI - Endothelial cells express normal cellular prion protein. PMID- 14531929 TI - Heparin-induced thrombocytopenia: pathogenesis and management. PMID- 14531932 TI - The feasibility of performing resistance exercise with acutely ill hospitalized older adults. AB - BACKGROUND: For older adults, hospitalization frequently results in deterioration of mobility and function. Nevertheless, there are little data about how older adults exercise in the hospital and definitive studies are not yet available to determine what type of physical activity will prevent hospital related decline. Strengthening exercise may prevent deconditioning and Pilates exercise, which focuses on proper body mechanics and posture, may promote safety. METHODS: A hospital-based resistance exercise program, which incorporates principles of resistance training and Pilates exercise, was developed and administered to intervention subjects to determine whether acutely-ill older patients can perform resistance exercise while in the hospital. Exercises were designed to be reproducible and easily performed in bed. The primary outcome measures were adherence and participation. RESULTS: Thirty-nine ill patients, recently admitted to an acute care hospital, who were over age 70 [mean age of 82.0 (SD= 7.3)] and ambulatory prior to admission, were randomized to the resistance exercise group (19) or passive range of motion (ROM) group (20). For the resistance exercise group, participation was 71% (p = 0.004) and adherence was 63% (p = 0.020). Participation and adherence for ROM exercises was 96% and 95%, respectively. CONCLUSION: Using a standardized and simple exercise regimen, selected, ill, older adults in the hospital are able to comply with resistance exercise. Further studies are needed to determine if resistance exercise can prevent or treat hospital-related deterioration in mobility and function. PMID- 14531933 TI - Visualisation and graph-theoretic analysis of a large-scale protein structural interactome. AB - BACKGROUND: Large-scale protein interaction maps provide a new, global perspective with which to analyse protein function. PSIMAP, the Protein Structural Interactome Map, is a database of all the structurally observed interactions between superfamilies of protein domains with known three dimensional structure in the PDB. PSIMAP incorporates both functional and evolutionary information into a single network. RESULTS: We present a global analysis of PSIMAP using several distinct network measures relating to centrality, interactivity, fault-tolerance, and taxonomic diversity. We found the following results: Centrality: we show that the center and barycenter of PSIMAP do not coincide, and that the superfamilies forming the barycenter relate to very general functions, while those constituting the center relate to enzymatic activity. Interactivity: we identify the P-loop and immunoglobulin superfamilies as the most highly interactive. We successfully use connectivity and cluster index, which characterise the connectivity of a superfamily's neighbourhood, to discover superfamilies of complex I and II. This is particularly significant as the structure of complex I is not yet solved. Taxonomic diversity: we found that highly interactive superfamilies are in general taxonomically very diverse and are thus amongst the oldest. Fault-tolerance: we found that the network is very robust as for the majority of superfamilies removal from the network will not break up the network. CONCLUSIONS: Overall, we can single out the P-loop containing nucleotide triphosphate hydrolases superfamily as it is the most highly connected and has the highest taxonomic diversity. In addition, this superfamily has the highest interaction rank, is the barycenter of the network (it has the shortest average path to every other superfamily in the network), and is an articulation vertex, whose removal will disconnect the network. More generally, we conclude that the graph-theoretic and taxonomic analysis of PSIMAP is an important step towards the understanding of protein function and could be an important tool for tracing the evolution of life at the molecular level. PMID- 14531934 TI - Liver abscess secondary to a broken needle migration--a case report. AB - BACKGROUND: Perforation of gut by sharp metallic objects is rare and rarer still is their migration to sites like liver. The symptoms may be non-specific and the discovery of foreign body may come as a radiological surprise to the unsuspecting clinician since the history of ingestion is difficult to obtain. CASE REPORT: A unique case of a broken sewing needle in the liver causing a hepatic abscess and detected as a radiological surprise is presented. The patient had received off and on treatment for pyrexia for the past one year at a remote primary health center. Exploratory laparotomy along with drainage of abscess and retrieval of foreign body relieved the patient of his symptoms and nearly one-year follow up reveals a satisfactory recovery. CONCLUSION: It is very rare for an ingested foreign body to lodge in the liver and present as a liver abscess. An ultrasound and a high clinical suspicion index is the only way to diagnose these unusual presentations of migrating foreign bodies. The management is retrieval of the foreign body either by open surgery or by percutaneous transhepatic approach but since adequate drainage of the abscess and ruling out of a fistulous communication between the gut and the liver is mandatory, open surgery is preferred. PMID- 14531935 TI - Effects of endothelin and nitric oxide on organ injury, mesenteric ischemia, and survival in experimental models of septic shock. AB - The development of potent drugs to treat cardiopulmonary failure in sepsis, such as antibiotics and new immunomodulatory therapeutic approaches have not prevented sepsis from being a major health problem. Dysfunction of the vascular endothelium is an early event in septic shock. The recognition of endothelium-derived substances, such as nitric oxide and endothelin, important mediators of systemic inflammatory response syndrome, led to the proposal that pharmacological inhibition of nitric oxide and endothelin production could represent a useful strategy in the treatment of septic shock. Splanchnic ischemia and translocation of endotoxin from the gut to the circulation contributes significantly to the high mortality rate in sepsis-related syndromes. This vasoconstriction in the splanchnic circulation can be partially blocked by inducible nitric oxide synthase inhibitor aminoguanidine or endothelin receptor antagonist bosentan in experimental models of septic shock. It can be suggested that endothelin and nitric oxide may affect survival. Although septic shock is a highly complex pathophysiological state, the course of septic shock has different phases with different characteristics which need different (special) treatment strategy. The inhibition of nitric oxide production during hyperdynamic, earlier phase of sepsis combined with the blockade of endothelin receptors at a later stage during the hypodynamic, late phase appears to be a novel promising strategy for the therapy of septic shock. The aim of this review is to discuss the role of nitric oxide and endothelin in sepsis and the potential therapeutic implications of blockade of nitric oxide and endothelin as a target in treatment of human septic shock. Briefly the importance of timing of intervention is also emphasized. PMID- 14531936 TI - Synergism interaction between arachidonic acid by 5-hydroxytryptamine in human platelet aggregation is mediated through multiple signalling pathways. AB - AIM: To examine the signalling mechanisms involved in the synergistic interaction of 5-hydroxytryptamine (5-HT) and arachidonic acid (AA) in human platelet aggregation. METHODS: Blood was obtained from healthy human subjects, mixed with 3.8 % sodium citrate (9:1), and centrifuged to prepare platelet rich plasma (PRP). Aggregation was monitored using a Dual-channel Lumi-aggregometer. The agonist-induced influx of Ca2+ was measured using Fura-2 AM. TXA2 formation was studied using radiochemical method. RESULTS: Subthreshold concentration of 5-HT (2 micromol/L) potentiated the effect of low dose of AA (0.2 mmol/L) in human platelets. This synergistic effect was blocked by 5-HT2 receptor antagonist (methysergide IC(50)=5.2 nmol/L; cyproheptadine IC(50)=0.6 nmol/L), and thromboxane A2 receptor antagonist (SQ 29 548; IC(50)=30 nmol/L), showing that the effect is receptor-mediated. To examine the down-stream signalling pathways, we found that such an interaction was inhibited by calcium channel blockers (diltiazem; IC(50)=3 micromol/L and verapamil; IC(50)=5 micromol/L), phospholipase C (PLC) inhibitor (U73122; IC50=4 micromol/L), cyclooxygenase inhibitor, (indomethacin; IC(50)=0.2 micromol/L) and mitogen-activated protein (MAP) kinase inhibitor (PD98059; IC(50)=3 micromol/L). The effect was also inhibited by a specific tyrosine light chain kinase (TLCK) inhibitor, herbimycin A with IC(50) value of 5 micromol/L. Pretreatment of platelet with 5-HT and AA induced rise in intracellular calcium and this effect was blocked by verapamil. CONCLUSION: The synergism between 5-HT and AA in platelet aggregation involves activation of PLC/Ca2+, COX, and MAP kinase pathways. PMID- 14531937 TI - Spatiotemporal relationships among D-serine, serine racemase, and D-amino acid oxidase during mouse postnatal development. AB - AIM: To elucidate the spatiotemporal relationships among D-serine, serine racemase, and D-amino acid oxidase (EC 1.4.3.3; DAO) in mouse cortex, striatum, cerebellum, heart, lung, liver, spleen, kidney, and skeletal muscle during mouse postnatal development. METHODS: The transcription levels of serine racemase and DAO were assayed by reverse transcription-polymerase chain reaction (RT-PCR). The protein levels of serine racemase were examined by Western blot. DAO activities were assayed by colorimetric method. D-serine was measured by HPLC. RESULTS: In cortex, striatum, and cerebellum, free D-serine increased drastically after birth and coincided well with the increase of serine racemase expression. However, among the 9 tissues examined, DAO activities were detected only in cerebellum and kidney. During the 3rd week, DAO activity in cerebellum and kidney increased dramatically, which concurred with the drastic decline of D-serine content in these tissues. On the other hand, while D-serine and serine racemase fall to trace level in cerebellum and kidney at the 3rd weekend, DAO activities in these tissues increased continuously. CONCLUSION: The free D-serine is mainly synthesized by serine racemase. However, novel mechanisms might be involved in D serine deposition in mouse tissues with high level of D-serine and no detectable DAO activity such as cortex and striatum. DAO in cerebellum and kidney might have other physiological functions in addition to degrading D-amino acid. PMID- 14531938 TI - Expression of core binding factor alpha1 up-regulated by IGF-I, GM-CSF, and EGF through MAPK pathway in MC3T3-E1 and C2C12 cells. AB - AIM: To study the regulating function and mechanism of insulin-like growth factor I (IGF-I), granulocyte-macrophage colony-stimulating factor (GM-CSF), and epidermal growth factor (EGF) on murine core binding factor alpha1 (Cbfalpha1) gene expression. METHODS: Luciferase reporter gene method and RT-PCR technique were used to examine the effects of these growth factors on the promoter activity and mRNA expression of Cbfalpha1 gene in MC3T3-E1 and C2C12 cells. RESULTS: IGF-I (from 1 nmol/L to 1 micromol/L), GM-CSF (100 nmol/L), and EGF (1 micromol/L) increased the luciferase expression in MC3T3-E1 cells (P<0.05). And mitogen activated protein kinase (MAPK) inhibitor, PD 98059 (10 micromol/L), completely blocked IGF-1, GM-CSF, and EGF-induced expression of Cbfa1 promoter activity (P<0.01). In C2C12 cells, IGF-I (from 1 nmol/L to 10 micromol/L), GM-CSF (100 nmol/L and 1 micromol/L), and EGF (100 nmol/L) enhanced the expression of luciferase reporter plasmid driven by mCbfalpha1 promoter (P<0.05). Addition of PD 98059 also blocked the stimulatory effects of these growth factors on Cbfalpha1 promoter activity (P<0.01). Moreover, Cbfalpha1 mRNA expression was significantly increased after treatment with IGF-I (1 nmol/L, 100 nmol/L), GM-CSF (100 nmol/L, 1 micromol/L), and EGF (1 micromol/L, 100 nmol/L) in MC3T3-E1 and C2C12 cells, respectively (P<0.05). These stimulatory effects of IGF-I, GM-CSF, and EGF on Cbfalpha1 mRNA expression were abolished by PD 98059. CONCLUSION: IGF I, GM-CSF, and EGF could increase the promoter activity and the mRNA expression of murine Cbfalpha1 gene in MC3T3-E1 and C2C12 cells. These stimulatory effects might be mediated by activating the intracellular MAPK-dependent signaling pathway. PMID- 14531939 TI - Expression of human alpha-galactosidase leads to reduction of major xenoepitope Galalpha(1,3) Gal in NIH 3T3 cell. AB - AIM: To examine the effects of the expression of alpha-galactosidase on the expression of the major xenoepitope Galalpha(1,3) Gal (G antigen) in NIH 3T3 cell. METHODS: The expression levels of G antigen and H antigen and binding of human natural antibodies (IgG and IgM) and complement (C3c) to NIH 3T3 cells were analyzed by flow cytometry. Western blot was employed to further determine the expression of glycoproteins of G antigen. Cytolysis assay with normal human serum was performed by MTT assay. RESULTS: In transfectants, Western blot showed that the binding of human IgG to glycosylated proteins located on the cell membrane was decreased, even abrogated totally. Together with the reduced binding of Gs IB4 (Griffonia simplicifolia) to transfectants, the stable expression of human alpha-galactosidase effectively inhibited Galalpha(1,3) Gal, Gal epitope synthesis in NIH 3T3 cell. As a result, the xenoreactivities of human IgG, IgM, and C3c were reduced by 73.4 %, 22.3 % and 47.9 %, respectively, while the cell lysis mediated by human XNA and complements was decreased by 42.4 %. CONCLUSION: The stable expression of human alpha-galactosidase in NIH 3T3 cell strongly inhibits the expression of Gal epitopes, resulting in abrupt reduction in xenorejection induced by human serum. PMID- 14531940 TI - Increase in drug-induced seizure susceptibility of transgenic mice overexpressing GABA transporter-1. AB - AIM: The changes of seizure susceptibility of transgenic mice overexpressing GABA transporter-1 (GAT-1) were studied to clarify the possible role of GABAergic transmission in epileptogenesis. METHODS: Seizures were induced by intraperitoneal administration of pentylenetetrazol (PTZ), picrotoxin (PIC), or kainic acid (KA) respectively. The anticonvulsant effect of ethyl nipecotate was tested by its intraperitoneal injection 15 min before the administration of the epileptogenic agents. RESULTS: The percentages of occurrence of clonic seizures induced by PTZ 45 mg/kg, PIC 2.5 mg/kg, or KA 20 mg/kg in GAT-1 transgenic mice were 88.9 %, 100 %, and 83.3 % respectively, whereas those in control C57BL/6J mice were 42.9 %, 57.1 %, and 33.3 %. The percentages of occurrence of tonic seizures induced by PTZ 45 mg/kg, PIC 2.5 mg/kg, or KA 20 mg/kg in transgenic mice were 88.9 %, 100 %, and 83.3 % respectively, and whereas those in control mice were 28.6 %, 42.9 %, and 16.7 %. The latencies of both clonic and tonic seizures onset in transgenic mice were markedly shortened compared with those in control animals. The results indicated that GAT-1 transgenic mice showed increased susceptibility to seizures induced by the anti-GABAergic convulsive drugs (PTZ, PIC), as well as glutamic receptor agonist (KA). Ethyl nipecotate, inhibitor of GAT-1, inhibited PTZ-induced seizures in both GAT-1 transgenic and C57BL/6J mice. The incidence of seizures was decreased after the application of ethyl nipecotate, and the latencies to the onset of clonic or tonic seizures were also prolonged. CONCLUSION: The increase in seizure susceptibility of transgenic mice over-expressing GAT-1 is an evidence for involvement of GABAergic transmission in epileptogenesis, and this transgenic mouse might be a useful animal model for study on the role of GABAergic transmission in epileptogenesis. PMID- 14531941 TI - Cytoprotective effect is one of common action pathways for antidepressants. AB - AIM: To explore the possible common action mechanism of antidepressants. METHODS: The cell viability was detected by MTT assay. The intracellular Ca2+ concentration ([Ca2+]i) was measured by Fura 2-AM fluorescence labeling assay. Using RT-PCR, the mRNA level of nerve growth factor (NGF) was also detected. RESULTS: High concentration of corticosterone (0.2 mmol/L) was incubated with PC12 cells to simulate the lesion state of brain neurons in depressive illness. Three main kinds of antidepressants used in clinic [(1) tricyclic antidepressants (TCAs), such as desipramine (DIM) 0.625-10 micromol/L; (2) selective serotonin reuptake inhibitors (SSRIs), such as fluoxetine (FLU) 0.625-10 micromol/L; (3) monoamine oxidase inhibitors (MAOIs), such as moclobemide (MOC) 2.5-40 micromol/L] protected cells from the lesion induced by corticosterone. While antipsychotic drug chlorpromazine or anxiolytic agent diazepam 0.4-50 micromol/L had no such effect. Moreover, DIM 1, 5 micromol/L or FLU 1, 5 micromol/L attenuated the [Ca2+]i overload induced by corticosterone 0.1 mmol/L for 48 h in PC12 cells. Furthermore, treatment with DIM or FLU 10 micromol/L for 48 h elevated the NGF mRNA expression in PC12 cells. CONCLUSION: Despite a remarkable structural diversity, the cytoprotective effect can be viewed as the common action pathway of the antidepressants. Moreover, attenuation of the intracellular Ca2+ overload and elevation of neurotrophic factor (such as NGF) expression is one of the mechanisms of cytoprotective effect of antidepressants. PMID- 14531942 TI - Effects of phenothiazine drugs on serum levels of apolipoproteins and lipoproteins in schizophrenic subjects. AB - AIM: To investigate the risk factors and clinical significance of blood-lipid metabolic disorder in schizophrenic patients caused by phenothiazine treatment for long term (from 1 month to 25 years). METHODS: Serum levels of apolipoprotein AI (apoAI), apolipoprotein B (apoB), high-density lipoprotein cholesterol (HDL C), triglyceride (TG), and total cholesterol (TC) were measured in 120 chronic schizophrenia patients, 50 vascular dementia, and 100 normal controls by the enzyme method and immune fluoroscopy turbidimetric method. RESULTS: The patients with schizophrenia and vascular dementia had significantly lower content of apoAI, HDL-C, and apoAI/apoB than those in normal control (P<0.01). Their apoB and TG levels were higher than the healthy control group (P<0.01). The TG contents in the negative group and the vascular dementia group were obviously higher than the positive group (P<0.01) while there was no marked difference between the TC levels in the three groups and the normal control group (P>0.05). CONCLUSION: The chronic schizophrenic patients have a blood-lipid metabolic disorder by long-term intake of phenothiazine drugs. It is suggested that the traditional treatment with antipsychotic should reformed, and that drugs of degrading lipid and coagulation should be used to prevent and reduce the risk factors causing the onset of cardiovascular and cerebrovascular diseases and delay the development of the disturbance of intelligence and dementia. PMID- 14531943 TI - Inhibitory effect of polypeptides from Chlamys farreri on UVB-induced apoptosis and DNA damage in normal human dermal fibroblasts in vitro. AB - AIM: To investigate the effect of polypeptide from Chlamys farreri (PCF) on ultraviolet B (UVB)-induced apoptosis and DNA damage in cultured normal human dermal fibroblasts. METHODS: MTT assay was used to measure the viability of cells. Measurements of apoptosis and cytosolic free [Ca2+]i were performed with flow cytometry. The comet assay was employed to detect DNA damage in individual cell. RESULTS: PCF (0.25 %-1%) greatly enhanced the proliferative capacity of cultured fibroblasts irradiated by UVB (1.176 x 10(-4) J/cm2) and markedly reduced apoptosis and the level of DNA damage in a concentration-dependent manner. Meanwhile, PCF could decrease the cytosolic free [Ca2+]i (P<0.01, compared with UVB model). CONCLUSION: The inhibitory effect of PCF on UVB-induced photoaging is due to enhanced abating of UVB-injured DNA and UVB-induced apoptosis. Therefore, PCF can resist UV-induced aging development at the initiation stage. PMID- 14531944 TI - Pharmacokinetic-pharmacodynamic modeling of daurisoline and dauricine in beagle dogs. AB - AIM: To study the combined pharmacokinetic-pharmacodynamic (PK-PD) model of daurisoline and dauricine, and compare their effects on cardiac electrophsiology, blood pressure, and hemodynamics in beagle dogs. METHODS: The plasma drug concentration was determined by the reversed-phase HPLC method and the effects on cardiac and hemodynamics were recorded by polygraph. The pharmacokinetic and PK PD model parameters were calculated. RESULTS: The pharmacokinetics were best fitted to a two-compartment open model, and the relationship between effect and effect compartment concentration of both drugs could be represented by the sigmoid-E(max) model. There were no significant differences in main pharmacokinetics and PK-PD parameters between the two drugs. CONCLUSION: No statistically different kinetic disposition characteristics and potencies of inhibitory effects on myocardial function of daurisoline and dauricine were found in beagle dogs. PMID- 14531945 TI - BmTx3B, a novel scorpion toxin from Buthus martensi Karsch, inhibits delayed rectifier potassium current in rat hippocampal neurons. AB - AIM: To examine the effect of BmTx3B, a novel short-chain peptide isolated from the venom of Asian scorpion Buthus martensi Karsch, on voltage-gated potassium channels. METHODS: Two types of voltage-dependent potassium currents were recorded from dissociated hippocampal neurons of neonatal rat in whole-cell voltage-clamp mode, and separated based upon their kinetic properties. RESULTS: BmTx3B (10-100 micromol/L) selectively inhibited the delayed rectifier potassium current (I(K)), without affecting the fast transient potassium current (I(A)). The inhibition of the peptide on I(K) was reversible, concentration-dependent and voltage-independent. BmTx3B did not affect the steady-state activation and inactivation kinetics of the current. CONCLUSION: The short-chain scorpion peptide BmTx3B selectively blocked the delayed rectifier potassium channel. PMID- 14531946 TI - Inhibitory effect of C-type natriuretic peptide on spontaneous contraction in gastric antral circular smooth muscle of rat. AB - AIM: To investigate the effect of natriuretic peptides on gastric motility and its possible mechanism. METHODS: Spontaneous contraction of gastric antral circular muscle of rats was recorded by four channel physiograph. The concentration of cyclic guanosine monophosphate (cGMP) was measured by radioimmunoassay. The distribution of natriuretic peptide receptors (NPR) was analyzed by autoradiograph. RESULTS: NPR existed in different regions of rat stomach and its density was the largest in gastric antrum. Atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and C-type natriuretic peptide (CNP) all inhibited the spontaneous contraction of gastric antral circular smooth muscle. Among them, the inhibitory effect of CNP on the spontaneous contraction was the most potent and exhibited a dose-dependent manner. CNP-induced inhibition was diminished by LY83583 (a kind of inhibitor of guanylate cyclase) and potentiated by zaprinist (a kind of inhibitor of cGMP sensitive phosphoesterase). CNP markedly enhanced the concentration of cGMP in antral smooth muscle. The inhibitory effect of CNP on spontaneous contraction was also inhibited by tetraethylammonium (a nonselective potassium channel blocker, TEA). CONCLUSION: The distribution density of NPR is the most in gastric antrum. CNP significantly inhibits gastric motility in rat gastric antrum. The inhibitory effect occurs via a cGMP dependent pathway and potassium channel is also involved in the relaxation induced by CNP in gastric antrial circular smooth muscle of rat. PMID- 14531947 TI - Protective effects of lovastatin on vascular endothelium injured by low density lipoprotein. AB - AIM: To examine protective effects of lovastatin, an inhibitor of 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase, on endothelial dysfunction induced by a single intravenous injection of natural low density lipoprotein (n-LDL) and analyze the possible action mechanism of lovastatin. METHODS: Rats were treated by intraperitoneal injection with lovastatin at dose of 2 or 4 mg/kg body weight once daily for 7 d, and on d 6 a single injection of n-LDL 4 mg/kg was given by sublingual vein. Forty eight hours after injection of n-LDL, the descending thoracic aorta of rats was taken. Acetylcholine (ACh)-induced endothelium dependent relaxation (EDR) and sodium nitroprusside (SNP)-induced endothelium independent relaxation of aortic rings were examined in vitro. Malondialdehyde (MDA), nitric oxide (NO), and activity of superoxide dismutase (SOD) as well as level of lipid in serum were analyzed. RESULTS: A single injection of n-LDL inhibited ACh-induced EDR compared with normal control group (maximal relaxation rate: 69.5 %+/-1.2 % vs 91.0 %+/-1.2 %, P<0.05), decreased serum NO level [(7.0+/ 0.5) micromol/L vs (11.2+/-0.9) micromol/L, P<0.05] and serum SOD activity [(371+/-16) kNU/L vs (405+/-18) kNU/L, P<0.05] and elevated serum MDA level [(5.4+/-0.5) micromol/L vs (3.0+/-0.8) micromol/L, P<0.05]. Compared with n-LDL treated group, lovastatin 2 and 4 mg/kg increased EDR( maximal relaxation rate 82.9 %+/-0.5% and 83.7 %+/-0.7 % vs 69.5 %+/-1.2 %, P<0.05) and elevated NO level [(11.0+/-0.7) and (11.2+/-0.8) micromol/L vs (7.0+/-0.5) micromol/L, P<0.05], increased SOD activity [(402+/-15) and (408+/-25) kNU/L vs (371+/-16) kNU/L, P<0.05], and reduced serum MDA level [(3.3+/-0.6) and (3.5+/-0.4) micromol/L vs (5.4+/-0.5) micromol/L, P<0.05]. But sodium nitroprusside-induced endothelium independent relaxation and the level of serum lipid in both saline+LDL group and lovastatin-treated group had no marked alteration. CONCLUSION: Lovastatin was able to protect vascular endothelium from dysfunction induced by a single injection of n-LDL. PMID- 14531948 TI - Effects of Ginkgo biloba extract and tanshinone on cytochrome P-450 isozymes and glutathione transferase in rats. AB - AIM: To investigate the effects of Ginkgo biloba extract (GbE) and tanshinone (Tan) on cytochrome P450 (CYP) isozymes and glutathione transferase (GT) in rats. METHODS: Several CYP-dependent reactions were monitored in liver and kidney microsomes of male rats treated ig with GbE and Tan daily for 10 d. The activity of GT, the levels of malondialdehyde (MDA) and nitric oxide (NO) in the tissues were also determined. RESULTS: CYP1A1, 1A2, and 2B1 activities in the liver were all significantly increased (2-9.5 fold) by pretreatment with GbE or Tan (P<0.01). An induction (1.4 fold) of CYP 2E1 activity was observed at the higher dose of GbE treatment (P<0.01), but a reduction (1.9 fold) after Tan administration (P<0.01). Whereas GbE could induce CYP3A (1.6 fold) (P<0.01) but Tan had no effects. Furthermore, the activities of CYP 1A1 (5.6-8.9 fold) and 1A2 (2.6 fold) in the kidney were induced by GbE (P<0.01). The activity of GT in rat liver receiving Tan was significantly increased (P<0.05) and a dramatic reduction in the activity of GT in the kidney was observed in the GbE-treated group (P<0.01). In addition, the GbE treatment markedly decreased the levels of MDA and NO in the tissues of rats (P<0.01). CONCLUSION: The modulation of CYP isozymes by GbE and Tan may result in altered metabolism of coadministered drugs. In addition, GbE is an active antioxidant and nitric oxide inhibitor in vivo. PMID- 14531949 TI - Coincidental increase of leukotriene B4 between cerebral cortex and lung tissue of sensitized rats. AB - AIM: To explore the changes of leukotrienes (LT) in cerebral cortex and lung tissues in ovalbumin-induced rat asthma model and effects of different anti asthma drugs on the changes. METHODS: Aerosol antigen-induced changes of inflammation in bronchoalveolar lavage fluids (BALF), pulmonary and brain histologic section in sensitized rats were investigated. Changes of LTB4 and LTC4 in lung and cerebral cortex homogenates were analyzed by reverse-phase high performance liquid chromatography (RP-HPLC). RESULTS: The number of inflammatory cells in BALF and the score of lung and brain histological examination from antigen- challenged rats were significantly higher than that from control group (P<0.05). Dexamethason (DXM, 0.5 mg/kg, ip) and ketotifen fumarate (KF, 5 mg/kg, ig) markedly reduced total leukocyte number in BALF, and inhibited eosinophil accumulation, reduced the infiltration of eosinophils, and improved mucous edema and epithelial lesion of bronchi and bronchioles. In addition, RP-HPLC results shown LTB4 in lung and cerebral cortex homogenates were increased in antigen challenged rats [(4.1+/-2.4) ng/g and (1.5+/-0.9) ng/g, respectively] compared with control group [(1.55+/-0.21) ng/g and (0.7+/-0.3) ng/g, respectively, P<0.05], both DXM (0.5 mg/kg, ip) and KF (5 mg/kg, ig) reduced LTB4 amount in lung[(1.4+/-0.6) ng/g and (1.8+/-0.7) ng/g] and cerebral cortex homogenates [(0.5+/-0.4) ng/g and (0.7+/-0.4) ng/g] in asthma rats. LTC4 content in lung homogenates in asthma rats was increased compared with control group [(1.9+/-0.9) ng/g and (0.5+/-0.3) ng/g, respectively] (P<0.05), but it has no change in cerebral cortex homogenates. DXM (0.5 mg/kg, ip) and KF (5 mg/kg, ig) reduced LTC4 amount in lung homogenates in asthma rats [(0.8+/-0.6) ng/g and (1.0+/-0.3) ng/g, respectively] (P<0.05). CONCLUSION: The results indicate there is coincidental increase of LTB4 between central nervous system and lung tissues in asthma rats. DXM and KF can inhibit the change. PMID- 14531950 TI - Nimesulide inhibits tumor growth in mice implanted hepatoma: overexpression of Bax over Bcl-2. AB - AIM: To investigate whether nimesulide could suppress tumor growth and induce apoptosis in implanted hepatoma mice and to explore the molecular mechanisms. METHODS: Male mice received nimesulide 10 mg/kg, 20 mg/kg, and 40 mg/kg ig daily for 21 d. Electron microscopy (EM), flow cytometry (FCM), DNA ladder, radioimmunoassay (RIA), and Western blot analysis were employed to investigate effect of nimesulide on mice hepatoma and the related molecular mechanisms. RESULTS: Nimesulide inhibited the growth of hepatoma (from 14 % to 62 %) and elicited typical apoptotic morphologic changes. The DNA ladder of high dose nimesulide was more clearly observed and apoptotic rate was 51.3 %+/-1.5 %. Nimesulide also decreased cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE2) and Bcl-2 expression, while increased the level of Bax protein. CONCLUSION: Nimesulide suppresses tumor growth and induces apoptosis by inhibiting COX-2 and PGE2 expression, which may be related to overexpression of Bax over Bcl-2. PMID- 14531951 TI - Putative caveolin-binding sites in SARS-CoV proteins. AB - AIM: To obtain the information of protein-protein interaction between the SARS CoV proteins and caveolin-1, identify the possible caveolin-binding sites in SARS CoV proteins. METHODS: On the basis of three related caveolin-binding motifs, amino acid motif search was employed to predict the possible caveolin-1 related interaction domains in the SARS-CoV proteins. The molecular modeling and docking simulation methods were used to confirm the interaction between caveolin-1 and SARS-CoV proteins. RESULTS: Thirty six caveolin-binding motifs in the SARS-CoV proteins have been mapped out using bioinformatics analysis tools. Molecular modeling and simulation have confirmed 8 caveolin-binding sites. These caveolin binding sites located in replicase 1AB, spike protein, orf3 protein, and M protein, respectively. CONCLUSION: Caveolin-1 may serve as a possible receptor of the SARS-CoV proteins, which may be associated with the SARS-CoV infection, replication, assembly, and budding. PMID- 14531952 TI - Antigen-specific T cells in rheumatoid arthritis. AB - There is considerable evidence of a key role for CD4+ T cells in the pathogenesis of rheumatoid arthritis. Several attractive candidate antigens, mostly joint specific, have been studied, but information regarding T cell responses to these antigens in patients is limited and occasionally contradictory. Novel reagents (such as major histocompatibility complex and peptide tetramers) and sensitive techniques (such as intracellular cytokine staining) will aid in future studies to identify antigen-specific T cells. In addition, a new animal model of inflammatory arthritis has recently provided new perspective to the study of rheumatoid arthritis by drawing attention to systemic self-antigens as targets of autoimmunity and anti-self antibodies as markers of T cell activity and effectors of disease. PMID- 14531954 TI - Lessons learned in the use of tumor necrosis factor-alpha inhibitors in the treatment of rheumatoid arthritis. AB - Tumor necrosis factor-alpha (TNFa) plays a central role in rheumatoid arthritis (RA) pathogenesis. There are currently three available anti-TNFa agents for the treatment of RA--adalimumab, etanercept, and infliximab. These targeted therapies have select advantages over traditional disease-modifying antirheumatic drugs (DMARDs), agents that have long been the mainstay of RA treatment. Compared with conventional DMARDs, TNFa inhibitors display a rapid onset of action and have shown a significant effect in retarding the radiographic joint destruction that often characterizes RA disease progression. Although anti-TNFa drugs represent an important advance in RA treatment, postmarketing reports of serious infections, as well as other adverse events, highlight the need for continued postmarketing vigilance with the use of these agents. This review evaluates the unique attributes of the available TNFa inhibitors, focusing specifically on recent reports providing important insight into the understanding of drug-related efficacy and toxicity. PMID- 14531953 TI - B lymphocytes and systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease characterized by B cell hyperactivity in association with autoantibodies, most prominently those directed to components of the cell nucleus. The source of the antigens that drive B cell responses in SLE is unknown, although recent studies suggest mechanisms by which the self-antigens become immunogenic and stimulate responses. Among these mechanisms, abnormalities in the generation of apoptotic cells or their clearance may increase the availability of nuclear antigens to drive responses. In addition, autoantibody crossreactivity may promote induction of responses to disparate antigens, foreign and self, and enable a single autoantibody to cause disease by crossreactive binding. In addition to reflecting increased exposure to self-antigen, autoantibody responses in SLE may result from abnormalities in B cell signaling and regulation by cytokines. New approaches to therapy aim to abrogate autoantibody production by targeting specific steps in B cell activation, including blockade of T cell costimulation. PMID- 14531955 TI - Atherosclerotic cardiovascular disease in rheumatoid arthritis. AB - The past 3 years have seen a remarkable growth in the interest of cardiovascular disease in rheumatoid arthritis. There have been studies published documenting an increased incidence and prevalence of cardiovascular conditions in patients with rheumatoid arthritis compared with individuals without rheumatoid arthritis. There has also been interest in the occurrence of cardiovascular risk factors in rheumatoid arthritis and in the role of antirheumatic therapy, including cyclooxygenase-2 selective nonsteroidal anti-inflammatory drugs, methotrexate, corticosteroids, and tumor necrosis factor inhibitors. A number of studies using noninvasive means to detect atherosclerosis have shown that patients with rheumatoid arthritis may be prone to atherosclerosis. This information should be important to physicians who provide care to patients with rheumatoid arthritis, given the difficulty of recognizing cardiovascular signs and symptoms among patients with the disease. PMID- 14531956 TI - Early rheumatoid arthritis: toward tailor-made therapy. AB - Therapeutic possibilities for the treatment of early rheumatoid arthritis (RA) have expanded largely. New treatment modalities appear very effective with respect to relevant outcomes, such as radiographic progression. At the same time, the costs of disease-modifying antirheumatic drugs (DMARDs) have exponentially increased so that--given the rather high prevalence of RA--cost may become a limiting factor in the treatment of patients with RA. Therefore, there is a need to define the profile of those patients that should be treated with the most effective, and, unfortunately, the most costly, DMARDs. The authors describe herewith the heterogeneity of RA with respect to its most important outcomes, as well as the inability to predict those outcomes appropriately at the individual patient level. This heterogeneity of RA is not acknowledged in the modern landmark clinical trials that the authors base therapeutic decisions on, and the external validity of those trials is at stake. In this article, the authors discuss the consequences of the heterogeneity of RA in light of the perceived lack of external validity of evidence-generating landmark trials. The authors propose the following solutions to overcome this discrepancy: 1) earlier recognition of RA, and 2) appropriate prediction of treatment efficacy, because the most challenging scientific efforts may be taken in the near future in order to arrive at a tailor-made therapy for every individual presenting with RA. PMID- 14531957 TI - The laryngeal and esophageal manifestations of Sjogren's syndrome. AB - Laryngopharyngeal reflux (LPR) is the syndrome caused by the backflow of gastric contents into the upper aerodigestive tract. Acid and pepsin in the pharynx, larynx, oral cavity, and trachea have been associated with dysphonia, chronic cough, reactive airway disease, middle ear effusion, throat pain, excessive throat mucus, postnasal drip, dental caries, and laryngeal cancer. The symptoms of LPR frequently occur in the absence of heartburn and esophagitis, and, thus, the diagnosis may be elusive. Individuals with Sjogren's syndrome are predisposed to reflux, and a high index of suspicion for LPR must be maintained in all individuals with the disease. This manuscript describes the laryngeal, pharyngeal, and esophageal manifestations of reflux in patients with Sjogren's syndrome and reviews state-of-the-art diagnostic and treatment strategies. PMID- 14531958 TI - The oral component of Sjogren's syndrome: pass the scalpel and check the water. AB - The labial salivary gland biopsy is a diagnostic test for the oral component of Sjogren's syndrome (SS) that has been the subject of controversy and re examination for many years. Despite multiple recent challenges to the significance of this test, when correctly done, it remains one of the most informative, specific, and technically simple tests available for the oral component of SS. Because of compromised salivary gland function, patients with SS are at risk for dental caries. Within the past decade, a paradigm shift has occurred within the field of caries research. The caries process was previously thought irreversible once initiated. Research has shown that the "early" carious lesion can be remineralized. Thus, the "early" carious lesion may be prevented and even repaired. The process of remineralization requires appropriate conditions to occur, and one of those conditions is pH. Because water is often the preferred wetting agent/beverage for patients with SS, the data show that, with regard to pH, not all waters are created equal. PMID- 14531959 TI - Recent advances in understanding molecular mechanisms in the pathogenesis and antibody profile of Sjogren's syndrome. AB - Sjogren's syndrome is a chronic autoimmune and rheumatic disorder of the mucous membranes caused by a lack of proper exocrine secretions, with prominent sicca complaints. The molecular mechanisms of the pathogenesis are virtually unknown, although progress has been made with regard to chemokines, B cell activating factor, and apoptosis. A large number of autoantibodies have been reported in Sjogren's syndrome, some of which relate to impairment of glandular function. Sjogren's syndrome is a female-dominant disease with a late age of onset; most patients contract the disease at the age of 40 to 50 years. Lately, attention has been drawn to the effects of adrenopause in Sjogren's syndrome and on dehydroepiandrosterone and its intracrine metabolism in target tissues. This can influence the maintenance and remodeling of exocrine glands and may explain, in part, another important disease symptom--fatigue. PMID- 14531960 TI - Lymphoproliferation in autoimmunity and Sjogren's syndrome. AB - Sjogren's syndrome is a chronic inflammatory process involving primarily the exocrine glands. Its association with lymphoma is well documented. A low-grade marginal-zone lymphoma related to mucosa-associated lymphoid tissue is the most common lymphoid neoplasia in Sjogren's syndrome. Among all autoimmune diseases, Sjogren's syndrome is the best tool to clarify the multiple components of autoimmunity and lymphomatogenesis. Herewith, the authors review the literature and discuss the molecular, clinical, histopathologic, and therapeutic aspects of these tumors in Sjogren's syndrome. PMID- 14531962 TI - Chronic urticaria and systemic diseases. PMID- 14531961 TI - The genetics of primary Sjogren's syndrome. AB - Primary Sjogren's syndrome is an autoimmune disease characterized clinically by dryness of the eyes and mouth. The use of different classification criteria for primary Sjogren's syndrome has led to dramatically different estimates of prevalence and incidence. Despite this, several genetic and environmental factors are thought to play a role in the susceptibility to primary Sjogren's syndrome, as is the current conceptual formulation of the pathogenesis of many other autoimmune maladies. Primary Sjogren's syndrome appears a complicated polygenic disorder with many genes interacting with environmental factors. Similar to many other polygenic autoimmune rheumatic diseases, human leukocyte antigen associations have been reported and confirmed. Additionally, other non-human leukocyte antigen candidate genes have been reported to reveal association with primary Sjogren's syndrome, but, in general, these effects are not confirmed. The authors review the human leukocyte antigen and non-human leukocyte antigen genetic associations herewith, knowing that new technologies are providing access to the entire genome for association studies. No doubt a much more comprehensive description of the genetics of this disorder will soon emerge. PMID- 14531966 TI - Vocal cord dysfunction in children and adolescents. AB - Vocal cord dysfunction (VCD) is a nonorganic disorder of the larynx that involves unintentional paradoxical adduction of the vocal cords while breathing. The resultant symptoms can include dyspnea, chest tightness, cough, throat tightness, wheezing, or voice change. Most patients with VCD are female, and among adolescents and children, VCD tends to be triggered by exercise and is typically confused with exercise-induced asthma. Both gastroesophageal reflux disease (GERD) and psychiatric illness have been reported as having strong associations with VCD, although, to date, there is no evidence that either causes VCD. VCD often coexists with asthma, and should be suspected in any patient in whom asthma treatment fails. Confirming the diagnosis involves direct visualization of abnormal vocal cord motion, and this usually only occurs during symptoms. Adolescent athletes often require free running exercise challenge to reproduce their symptoms and confirm abnormal vocal cord motion laryngoscopically. The primary treatment for VCD involves a combination of patient education and speech therapy, and, in most cases, patients may resume their activities without significant limitation. PMID- 14531967 TI - Practice parameters in pediatric allergy. AB - The primary role of practice parameter/guidelines and other boundaries developed for the field of medicine is to improve the quality of patient care. Practice parameters/guidelines are also important for education, interaction with managed care and third-party payers, establishing appropriate variables for outcome assessment, reducing inappropriate variation in clinical practice, and resolving medical-legal issues. National specialty organizations, the American Medical Association, and government agencies have been actively involved in developing and promoting practice parameters. These documents provide a framework within which pediatricians can provide the best quality of care for their patients. PMID- 14531968 TI - The need for pediatric studies of allergy and asthma medications. AB - For many years, clinicians have accepted the fact that most medications do not have dosing guidelines for children younger than 12 years of age. Recently, there has been a great effort to correct this deficiency. With the introduction of the 1997 Food and Drug Administration Modernization Act, a provision was established to grant additional market exclusivity to pharmaceutical firms that performed the required studies that would lead to improved labeling of medications for children. This effort has resulted in a significant advance for the management of asthma and allergic disorders in children. Several allergy and asthma medications are now approved for use in children as young as 1 year of age, with studies currently being conducted in younger age groups. In this review, we discuss the background for this effort and the continuing impact it will have on the future management of allergy and asthma in children. PMID- 14531969 TI - Modification of provider behavior to achieve improved asthma outcomes. AB - Despite an abundance of scientific evidence supporting the use of guidelines, adherence to asthma practice guidelines by physicians generally is low, regardless of provider and patient characteristics. As a result, scientific information, obtained with great effort and at huge expense, is not being translated into clinical practice. To remedy this, we developed a disease management program that emphasizes alteration of provider behavior using operant conditioning. We did this by placing asthma educators in private offices for up to 8 weeks. The educators used a combination of problem-based learning, role modeling, and operant conditioning with positive reinforcement to affect behavior change. As a result of these behavior changes, by the end of 8 weeks the cost to treat asthma patients decreased, despite an increase in the cost of medications. We concluded that behavior-oriented programs targeted at provider offices can lead to improved asthma care while reducing costs. PMID- 14531970 TI - Development of an asthma disease management program in a children's hospital. AB - The incidence, morbidity, and mortality of asthma have been increasing at an alarming rate, making asthma the most common chronic illness of childhood. An asthma disease management program was developed to improve the care and management of patients with asthma--a comprehensive health care delivery model that was designed to improve the management of patients with asthma was designed and implemented. The goal of the program was to provide high-quality interventions for those children diagnosed with asthma. The asthma disease management program at Children's Mercy Hospital improved the care received, decreased costs, and improved the quality of life for those children with asthma. PMID- 14531972 TI - Molecular immunology and immunotherapy for chronic sinusitis. AB - The development of chronic sinusitis is a complex multifactorial process characterized by inflammation of nasal and sinus mucosa. Many studies have shown that the composition of the inflammatory substrate in chronic sinusitis is similar to that seen in allergic rhinitis and in the late-phase response to antigen challenge. Mononuclear cells, consisting of T and B lymphocytes and activated eosinophils, are prominent in the sinus mucosa of patients with chronic sinusitis, especially in atopic patients. Cellular recruitment and activation of the inflammatory infiltrate have been largely attributed to the effects of T(H)2 cytokines (namely interleukin -4, IL-5, IL-13, and granulocyte-macrophage colony stimulating factor). Current treatment of allergic chronic sinusitis consists of nasal corticosteroids and immunotherapy. A subgroup of steroid-insensitive patients demonstrates an overexpression of a variant of the glucocorticoid receptor (GR). Despite these advances, the management and treatment of chronic sinusitis is often fraught with failures and remains a frustrating task for both physician and patient. PMID- 14531974 TI - Endoscopic and imaging techniques in the diagnosis of chronic rhinosinusitis. AB - The definition of chronic rhinosinusitis globally applies to any patient who has nasal and/or sinus inflammatory disease for a period of more than 3 months, provided the patient has particular signs and symptoms. However, to treat chronic rhinosinusitis appropriately, specific etiologies must be diagnostically ruled in or out. Chronic rhinosinusitis can be complex, because there are several factors involved in its causation. Endoscopy and imaging can play a role in helping to lead to a more specific diagnosis, allowing more targeted treatment. PMID- 14531973 TI - The impact of rhinosinusitis on asthma. AB - The nasal cavity, paranasal sinuses, and lungs are considered separate organs of the respiratory tract. However, a growing body of evidence links the upper and lower airways. For example, the coexistence and impact of allergic and nonallergic rhinitis on asthma is now documented. In addition, inflammation of the nose (rhinitis) commonly is associated with inflammation of sinuses (sinusitis), as reflected in the term rhinosinusitis. In this paper, we review the impact of rhinosinusitis on asthma as it relates to the links between allergic and nonallergic rhinitis and asthma; viral upper respiratory tract infections and asthma; allergic and nonallergic infectious/inflammatory rhinosinusitis and allergic and nonallergic hyperplastic rhinosinusitis and asthma; and the aspirin-exacerbated respiratory disease syndrome and asthma. PMID- 14531976 TI - Confronting the new challenge in travel medicine: SARS. PMID- 14531975 TI - Superantigens and nasal polyps. AB - Nasal polyps represent an often severe T-cell-orchestrated eosinophilic upper airway disease with currently unknown pathogenesis, often associated with lower airway disease, such as asthma. Superantigens, predominantly derived from Staphylococcus aureus, are potent activators of T cells, induce the synthesis of IgE in B cells, and have direct effects on pro-inflammatory cells, such as eosinophils. IgE antibodies to S. aureus enterotoxins have been described in polyp tissue, linked to a local polyclonal IgE production and an aggravation of eosinophilic inflammation. Furthermore, such IgE antibodies have also been described in the sera of patients with asthma, and linked to severity of disease and steroid insensitivity. This review summarizes our current understanding of the possible role of S. aureus enterotoxins in chronic severe airway disease, such as nasal polyposis. PMID- 14531977 TI - Experience of severe acute respiratory syndrome in singapore: importation of cases, and defense strategies at the airport. AB - BACKGROUND: The importation of SARS was responsible for the outbreaks in Singapore, Hong Kong, Vietnam and Canada at a time when this new disease had not been identified. We report the incidence and impact of cases of SARS imported to Singapore between 25 February and 31 May 2003, and describe national measures to prevent further importation. METHODS: Information on imported cases of SARS and measures taken at entry points to Singapore was retrieved from the Ministry of Health and the Civil Aviation Authority of Singapore. RESULTS: Of the 6 imported cases, which all occurred before screening measures were implemented at the airport, only the first resulted in extensive secondary transmission. Of 442973 air passengers screened after measures were implemented, 136 were sent to a designated hospital for further SARS screening; none was diagnosed as having SARS. CONCLUSIONS: The SARS outbreak in Singapore can be traced to the first imported case. The absence of transmission from the other imported cases was probably a result of relatively prompt identification and isolation of cases, together with a low potential for transmission. New imported SARS cases therefore need not lead to major outbreaks if systems are in place to identify and isolate them early. Screening at entry points is costly, has a low yield and is not sufficient in itself, but may be justified in light of the major economic, social and international impact which even a single imported SARS case may have. PMID- 14531978 TI - Health-related challenges in United States Peace Corps Volunteers serving for two years in Madagascar. AB - BACKGROUND: The United States Peace Corps has nearly 8000 Volunteers (PCVs) working in more than 70 countries, predominantly in the developing part of the world. The volunteers are challenged by many new and unfamiliar health problems, as a result of change in living conditions during their service. The goal of this study was to assess the health problems of PCVs in Madagascar, and to use this information to evaluate the effectiveness of the current PCV health support program. METHODS: PCVs, finalizing their 2 years of service, were enrolled in the study. Data were retrieved from individual Peace Corps service medical records, the close of service (COS) physical evaluation, and a PCV questionnaire, which elicited additional self-reported health problems and concerns. RESULTS: Sixty nine PCVs took part in the study. Median age at COS was 24 years (95% CI 23-26 years), and the male/female ratio was 1 : 3.6 (15 males and 54 females). Skin, gastrointestinal and respiratory infections constituted the three most common reported health problems. Among the registered parasitic infections were ova and parasites in the stool (42.0%), confirmed and presumptive malaria (15.9%) and schistosomiasis (11.6%). Female PCVs more frequently reported dermatologic manifestations than male PCVs: postadolescent acne (48.2% vs. 13.3%) and telogen hair loss (33.3% vs. 6.7%). Amenorrhea, as the most commonly reported menstrual alteration during service, was reported by 15 (27.7%) female PCVs. CONCLUSIONS: The PCVs in Madagascar encounter a broad range of health-related problems, most of which are preventable infectious diseases. Reinforcement of the preservice personal health-training curriculum later during service is needed. Reported dermatologic and menstrual disturbances in female subjects may be associated with stress-related estrogen imbalance. More studies are warranted to assess the stress impact of long-term travel on the female physiology. PMID- 14531979 TI - Life and death on the Amazon: illness and injury to travelers on a South American expedition. AB - BACKGROUND: Commercial expeditions provide an opportunity for travelers to undertake various specialized travel to more adventurous destinations in the relative security of an expeditionary group. Little is known about the illnesses and injuries occurring on such expeditions. METHODS: From the commencement of the Blake Expedition in 2001, the expedition physician recorded all illnesses and injuries reported among the crew and also among the indigenous people encountered, while undertaking the expedition. The duration of the expedition was two and a half months, and involved travel by yacht, riverboat and jungle canoe or "bongo". The crew numbered 26 personnel: 24 men, including the expedition physician, and two women. RESULTS: Seventy-eight health problems were reported and recorded among 19 (73.1%) members of the expeditionary team. There was one death, but there were no other major incidents requiring emergency evacuation or hospitalization. Types of illness were largely those related to ear, nose and throat disease (15/78, 19%), injuries (12/87, 15%), bites and stings (12/87, 15%), and respiratory (10/78, 13%), dermatologic (9/78, 12%) and gastroenterologic conditions (7/78, 9%). A further 24 health problems were reported and recorded among 22 indigenous people who approached the expedition physician for treatment. Types of illness were largely related to dermatologic conditions (9/24, 38%), accidents and injuries (4/24, 17%), and malaria (4/24, 17%). CONCLUSIONS: This expedition was both noteworthy and newsworthy because of the death of the team leader, Sir Peter Blake, although the health problems encountered were largely similar to those reported for other expeditions. Tropical disease was uncommon. Adequate pre-trip preparation of expedition teams is considered important, and the inclusion of an expedition physician decreased the reliance on local health services, which are often scarce or absent on more remote-location expeditions. Adequate psychological preparation of expedition teams should also be included, to prepare expedition teams for unexpected outcomes, such as death or severe injury. PMID- 14531980 TI - Travel health information at commercial travel websites. AB - BACKGROUND: Internet travel purchases accounted for 10% of the travel industry revenue generated in 2001. To ensure that travelers remain healthy during excursions to developing countries, travel health information needs to be available at commercial travel websites. We evaluated the current availability of travel health information at these websites. METHODS: The existence, adequacy and ease of access of the travel health information provided on commercial travel websites was assessed through a review of the top 25 airline and 20 discount travel websites. Each site was examined to determine whether it provided general information, such as jet lag, or international travel health information, such as malaria prophylaxis. We also assessed hyperlinks to external travel health information websites, such as the CDC, when provided. RESULTS: Travel health information was not available at 20 (44%) commercial travel websites, including 36% of airline and 55% of the discount travel websites. Twenty-eight percent of airline websites contained general information only, 8% links only, and 28% general and international information. Travel health information available at discount travel websites included 10% general only, 30% link only, and 5% general and international information. On average, it took three clicks to access travel health information. Keywords clicked to access travel health information frequently did not obviously refer to health. Each of the six travel health website links provided accurate vaccine and travel health information. However, several links lacked disease-specific maps and details of disease risk (i.e. seasonal and regional variations of malaria risk). CONCLUSIONS: Travel health information on commercial travel websites may be the only data available to travelers purchasing online. The information currently provided is generally inadequate. Ideally, commercial travel websites would provide uniform information that is accurate and easily accessible. Internationally recognized organizations should consider establishing guidelines for the information provided on commercial travel websites. PMID- 14531981 TI - Health risks for travelers to Lebanon. PMID- 14531982 TI - Imported Dengue in French University Hospitals: a 6-year survey. AB - Dengue fever (DF) is a mosquito-transmitted acute disease caused by any of four dengue flavivirus serotypes (DEN-1, DEN-2, DEN-3, and DEN-4) which is becoming a major public health problem in intertropical areas.1 DF is increasingly observed in febrile travelers returning from tropical areas,2 especially those returning from the Caribbean islands and Southeast Asia, but it is rarely diagnosed in travelers returning from Africa.3-8 Diagnosis is often serologic, and in most cases is not confirmed; virus isolation remains exceptional. In a retrospective study of 44 cases of imported DF diagnosed in France, we found that the epidemiologic, clinical and diagnostic characteristics of these cases were similar to those reported in other previous published studies; diagnosis was carried out with serology, and no virus isolation was reported. To draw the attention of physicians from nonendemic areas to the possible occurrence of dengue infection in febrile travelers from all tropical countries, including those in Africa, and to reinforce recommendations required to establish diagnosis with certainty, we report herein a prospective study from 1998-1999 which we compiled with our previously published data. PMID- 14531983 TI - Irrational prescribing in South Asia: a case of fluoroquinolone-associated phototoxicity. AB - Prescribing habits in South Asian countries have been subjected to some scrutiny.1-6 Most studies conclude that the quality of prescribing is poor, with overuse of antimicrobials and irrational use of fixed-dose combination therapy, particularly in the private sector.1 Prescriptions for multiple drugs are the rule rather than the exception, with up to seven items being prescribed for a single disease entity. Analgesics, anti-inflammatories and drugs of uncertain pharmacologic efficacy, such as vitamins, minerals and glucose water, are also frequently prescribed. PMID- 14531984 TI - Myiasis with Lund's fly (Cordylobia rodhaini) in travelers. AB - Myiasis is an infestation of human tissue by the larvae of certain flies. There are many forms of myiasis, including localized furuncular myiasis, creeping dermal myiasis and wound and body cavity myiasis.1 Cordylobia anthropophaga (the Tumbu fly) and Dermatobia hominis (the human botfly) are the most common causes of myiasis in Africa and tropical America respectively. The genus Cordylobia also contains two less common species, C. ruandae and C. rodhaini. The usual hosts of C. rodhaini are various mammals (particularly rodents), and and humans are accidentally infested. Figure 1 shows the life cycle of C. rodhaini, which occurs over 55 to -67 days.3 The female fly deposits her eggs on dry sand polluted with the excrement of animals or on human clothing. In about 3 days, the larva is activated by the warm body of the host, hatches and invades the skin. As the larva matures, it induces a furuncular swelling. In 12 to -15 days, the larva reaches a length of about 23 mm, exits the skin and falls to the ground to pupate. The adult fly emerges in 23 to -26 days, and the life cycle resumes. In humans, the skin lesion starts as a red papule that gradually enlarges and develops into a furuncle. In the center of the lesion an opening forms, through which the larva breaths and discharges its serosanguinous feces. The lesion is associated with increasing pain until the larva exits the skin. The disease is usually uncomplicated and self-limiting. PMID- 14531985 TI - Risk factors and characteristics of patients with acute Plasmodium falciparum malaria presenting to a private hospital network in South Africa. PMID- 14531986 TI - In-flight transmission of Severe Acute Respiratory Syndrome (SARS): a case report. PMID- 14531987 TI - Late symptomatic Plasmodium malariae relapse in the territory of the former Yugoslavia. PMID- 14531988 TI - Suspected allergy to artemether-lumefantrine treatment of malaria. PMID- 14531989 TI - Nodular lesion of the skin as primary cutaneous tuberculosis. PMID- 14531990 TI - Selected Bibliography. PMID- 14531992 TI - Increased genome instability in aging yeast. AB - In the September 26 issue of Science, McMurray and Gottschling (2003) report that aged yeast cells display high rates of loss of heterozygosity. Furthermore, they show that this reflects an impaired ability to correctly detect and repair DNA double-strand breaks. These results provide insights into how aging can engender genomic instability in eukaryotic cells. PMID- 14531993 TI - Bacterial vesicle formation as a mechanism of protein transfer to animals. AB - Gram-negative bacterial vesicle formation is a mechanism for specific secretion and transfer of a protein toxin to animals. This discovery should stimulate work on the mechanism of protein sorting into vesicles and the role of vesicles in bacterial pathogenesis. PMID- 14531994 TI - A polo match for Plk1. AB - New work by Elia et al. in this issue of Cell reveals the molecular basis of phosphopeptide recognition by the polo domain and the domain's dual function to promote substrate recognition by targeting the kinase to subcellular structures and to autoregulate the adjacent protein kinase catalytic domain. PMID- 14531995 TI - Does it take two to untangle? AB - The structural integrity of mitotic chromosomes is essential for proper chromatid segregation. In this issue of Cell, show that vertebrates contain two distinct condensin complexes, both of which are required for normal mitotic chromosome morphology. PMID- 14531999 TI - By binding SIRPalpha or calreticulin/CD91, lung collectins act as dual function surveillance molecules to suppress or enhance inflammation. AB - Surfactant proteins A and D (SP-A and SP-D) are lung collectins composed of two regions, a globular head domain that binds PAMPs and a collagenous tail domain that initiates phagocytosis. We provide evidence that SP-A and SP-D act in a dual manner, to enhance or suppress inflammatory mediator production depending on binding orientation. SP-A and SP-D bind SIRPalpha through their globular heads to initiate a signaling pathway that blocks proinflammatory mediator production. In contrast, their collagenous tails stimulate proinflammatory mediator production through binding to calreticulin/CD91. Together a model is implied in which SP-A and SP-D help maintain a non/anti-inflammatory lung environment by stimulating SIRPalpha on resident cells through their globular heads. However, interaction of these heads with PAMPs on foreign organisms or damaged cells and presentation of the collagenous tails in an aggregated state to calreticulin/CD91, stimulates phagocytosis and proinflammatory responses. PMID- 14532000 TI - Vesicle-mediated export and assembly of pore-forming oligomers of the enterobacterial ClyA cytotoxin. AB - The ClyA protein is a pore-forming cytotoxin expressed by Escherichia coli and some other enterobacteria. It confers cytotoxic activity toward mammalian cells, but it has remained unknown how ClyA is surface exposed and exported from bacterial cells. Outer-membrane vesicles (OMVs) released from the bacteria were shown to contain ClyA protein. ClyA formed oligomeric pore assemblies in the OMVs, and the cytotoxic activity toward mammalian cells was considerably higher than that of ClyA protein purified from the bacterial periplasm. The redox status of ClyA correlated with its ability to form the oligomeric pore assemblies. In bacterial cells with a defective periplasmic disulphide oxidoreductase system, the ClyA protein was phenotypically expressed in a constitutive manner. The results define a vesicle-mediated transport mechanism in bacteria, and our findings show that the localization of proteins to OMVs directly may contribute to the activation and delivery of pathogenic effector proteins. PMID- 14532001 TI - Formation of an endophilin-Ca2+ channel complex is critical for clathrin-mediated synaptic vesicle endocytosis. AB - A tight balance between synaptic vesicle exocytosis and endocytosis is fundamental to maintaining synaptic structure and function. Calcium influx through voltage-gated Ca2+ channels is crucial in regulating synaptic vesicle exocytosis. However, much less is known about how Ca2+ regulates vesicle endocytosis or how the endocytic machinery becomes enriched at the nerve terminal. We report here a direct interaction between voltage-gated Ca2+ channels and endophilin, a key regulator of clathrin-mediated synaptic vesicle endocytosis. Formation of the endophlin-Ca2+ channel complex is Ca2+ dependent. The primary Ca2+ binding domain resides within endophilin and regulates both endophilin-Ca2+ channel and endophilin-dynamin complexes. Introduction into hippocampal neurons of a dominant-negative endophilin construct, which constitutively binds to Ca2+ channels, significantly reduces endocytosis-mediated uptake of FM 4-64 dye without abolishing exocytosis. These results suggest an important role for Ca2+ channels in coordinating synaptic vesicle recycling by directly coupling to both exocytotic and endocytic machineries. PMID- 14532002 TI - Phosphorylation of RIM1alpha by PKA triggers presynaptic long-term potentiation at cerebellar parallel fiber synapses. AB - Presynaptic activation of protein kinase A (PKA) induces LTP in cerebellar parallel fiber synapses. Presynaptic LTP is known to require the active zone protein RIM1alpha, but the underlying induction mechanism remains unclear. We now show that PKA directly phosphorylates RIM1alpha at two sites. Using paired recordings from cultured cerebellar granule and Purkinje neurons, we demonstrate that LTP is absent in neurons from RIM1alpha KO mice but is rescued by presynaptic expression of RIM1alpha. Mutant RIM1alpha lacking the N-terminal phosphorylation site is unable to rescue LTP in RIM1alpha knockout neurons but selectively suppresses LTP in wild-type neurons. Our findings suggest that PKA mediated phosphorylation of the active zone protein RIM1alpha at a single N terminal site induces presynaptic LTP. PMID- 14532003 TI - A JNK-dependent pathway is required for TNFalpha-induced apoptosis. AB - Tumor necrosis factor (TNFalpha) receptor signaling can simultaneously activate caspase 8, the transcription factor, NF-kappaB and the kinase, JNK. While activation of caspase 8 is required for TNFalpha-induced apoptosis, and induction of NF-kappaB inhibits cell death, the precise function of JNK activation in TNFalpha signaling is not clearly understood. Here, we report that TNFalpha mediated caspase 8 cleavage and apoptosis require a sequential pathway involving JNK, Bid, and Smac/DIABLO. Activation of JNK induces caspase 8-independent cleavage of Bid at a distinct site to generate the Bid cleavage product jBid. Translocation of jBid to mitochondria leads to preferential release of Smac/DIABLO, but not cytochrome c. The released Smac/DIABLO then disrupts the TRAF2-cIAP1 complex. We propose that the JNK pathway described here is required to relieve the inhibition imposed by TRAF2-cIAP1 on caspase 8 activation and induction of apoptosis. Further, our findings define a mechanism for crosstalk between intrinsic and extrinsic cell death pathways. PMID- 14532004 TI - Proteasome-mediated degradation of p21 via N-terminal ubiquitinylation. AB - We examined the mechanism responsible for the degradation of p21, a negative regulator of the cell division cycle. We found that p21 proteolysis requires functional ubiquitin and Nedd8 systems. Ubiquitinylated forms of p21 and p21(K0), a p21 mutant missing all lysines, are detected in vivo and in vitro, showing that the presence of lysines is dispensable for p21 ubiquitinylation. Instead, the free amino group of the N-terminal methionine of p21 is a site for ubiquitinylation in vivo. Although wild-type p21 is more abundantly ubiquitinylated than p21(K0) mutant due to the presence of internal lysine residues, their rates of proteolysis are indistinguishable. These results demonstrate that proteasomal degradation of p21 is regulated by the ubiquitin pathway and suggest that the site of the ubiquitin chain is critical in making p21 a competent substrate for the proteasome. PMID- 14532005 TI - The molecular basis for phosphodependent substrate targeting and regulation of Plks by the Polo-box domain. AB - Polo-like kinases (Plks) perform crucial functions in cell-cycle progression and multiple stages of mitosis. Plks are characterized by a C-terminal noncatalytic region containing two tandem Polo boxes, termed the Polo-box domain (PBD), which has recently been implicated in phosphodependent substrate targeting. We show that the PBDs of human, Xenopus, and yeast Plks all recognize similar phosphoserine/threonine-containing motifs. The 1.9 A X-ray structure of a human Plk1 PBD-phosphopeptide complex shows that the Polo boxes each comprise beta6alpha structures that associate to form a 12-stranded beta sandwich domain. The phosphopeptide binds along a conserved, positively charged cleft located at the edge of the Polo-box interface. Mutations that specifically disrupt phosphodependent interactions abolish cell-cycle-dependent localization and provide compelling phenotypic evidence that PBD-phospholigand binding is necessary for proper mitotic progression. In addition, phosphopeptide binding to the PBD stimulates kinase activity in full-length Plk1, suggesting a conformational switching mechanism for Plk regulation and a dual functionality for the PBD. PMID- 14532006 TI - Structure of the mammalian mitochondrial ribosome reveals an expanded functional role for its component proteins. AB - The mitochondrial ribosome is responsible for the biosynthesis of protein components crucial to the generation of ATP in the eukaryotic cell. Because the protein:RNA ratio in the mitochondrial ribosome (approximately 69:approximately 31) is the inverse of that of its prokaryotic counterpart (approximately 33:approximately 67), it was thought that the additional and/or larger proteins of the mitochondrial ribosome must compensate for the shortened rRNAs. Here, we present a three-dimensional cryo-electron microscopic map of the mammalian mitochondrial 55S ribosome carrying a tRNA at its P site, and we find that instead, many of the proteins occupy new positions in the ribosome. Furthermore, unlike cytoplasmic ribosomes, the mitochondrial ribosome possesses intersubunit bridges composed largely of proteins; it has a gatelike structure at its mRNA entrance, perhaps involved in recruiting unique mitochondrial mRNAs; and it has a polypeptide exit tunnel that allows access to the solvent before the exit site, suggesting a unique nascent-polypeptide exit mechanism. PMID- 14532007 TI - Differential contributions of condensin I and condensin II to mitotic chromosome architecture in vertebrate cells. AB - The canonical condensin complex (henceforth condensin I) plays an essential role in mitotic chromosome assembly and segregation from yeast to humans. We report here the identification of a second condensin complex (condensin II) from vertebrate cells. Condensins I and II share the same pair of structural maintenance of chromosomes (SMC) subunits but contain different sets of non-SMC subunits. siRNA-mediated depletion of condensin I- or condensin II-specific subunits in HeLa cells produces a distinct, highly characteristic defect in chromosome morphology. Simultaneous depletion of both complexes causes the severest defect. In Xenopus egg extracts, condensin I function is predominant, but lack of condensin II results in the formation of irregularly shaped chromosomes. Condensins I and II show different distributions along the axis of chromosomes assembled in vivo and in vitro. We propose that the two condensin complexes make distinct mechanistic contributions to mitotic chromosome architecture in vertebrate cells. PMID- 14532010 TI - Seasonal trends in photosynthetic parameters and stomatal conductance of blue oak (Quercus douglasii) under prolonged summer drought and high temperature. AB - Understanding seasonal changes in photosynthetic parameters and stomatal conductance is crucial for modeling long-term carbon uptake and energy fluxes of ecosystems. Gas exchange measurements of CO2 and light response curves on blue oak leaves (Quercus douglasii H. & A.) were conducted weekly throughout the growing season to study the seasonality of photosynthetic capacity (Vcmax) and Ball-Berry slope (m) under prolonged summer drought and high temperature. A leaf photosynthetic model was used to determine Vcmax. There was a pronounced seasonal pattern in Vcmax. The maximum value of Vcmax, 127 micromol m(-2) s(-1), was reached shortly after leaf expansion in early summer, when air temperature was moderate and soil water availability was high. Thereafter, Vcmax declined as the soil water profile became depleted and the trees experienced extreme air temperatures, exceeding 40 degrees C. The decline in Vcmax was gradual in midsummer, however, despite extremely low predawn leaf water potentials (Psipd, approximately -4.0 MPa). Overall, temporal changes in Vcmax were well correlated with changes in leaf nitrogen content. During spring leaf development, high rates of leaf dark respiration (Rd, 5-6 micromol m(-2) s(-1)) were observed. Once a leaf reached maturity, Rd remained low, around 0.5 micromol m(-2) s(-1). In contrast to the strong seasonality of Vcmax, m and marginal water cost per unit carbon gain (partial partial differential E/ partial partial differential A) were relatively constant over the season, even when leaf Psipd dropped to -6.8 MPa. The constancy of partial partial differential E/ partial partial differential A suggests that stomata behaved optimally under severe water-stress conditions. We discuss the implications of our findings in the context of modeling carbon and water vapor exchange between ecosystems and the atmosphere. PMID- 14532011 TI - Improvement in growth and salt resistance of lemon (Citrus limon) trees by an interstock-induced mechanism. AB - Interstocks can reduce toxic ion accumulations in leaves of budded citrus trees, but the mechanism is not understood. We grew sour orange (Citrus aurantium L.; SO) seedlings, budded trees of 'Salustiano' orange (Citrus sinensis L. Osbeck; SAO) on SO, 'Verna' lemon (Citrus limon L. Burm. f; VL/SO) and interstock trees (VL/SAO/SO) in pots of sand watered with nutrient solution containing 5 (control) or 50 mM NaCl (saline treatment) for 12 weeks. Plants were harvested in six successive harvests and time trends in relative growth rate (RGR) and its components were estimated by fitting a Richards function regression to the harvest data. The VL/SAO/SO trees in saline conditions had higher mean RGR than VL/SO trees in control conditions. Increases in both net assimilation rate on a leaf mass basis (NARw) and leaf mass fraction (LMF) contributed equally to a twofold increase in RGR of VL/SAO/SO trees in saline conditions. In control conditions, the increase in RGR caused by the interstock had growth response coefficients of GRC(NARw) = 0.20 and GRC(LMF) = 0.80. Structural modifications specific leaf area, leaf area ratio and LMF-had a slight influence on the salt induced changes in RGR, whereas NARw had a large influence. Salinity decreased root mass fraction (RMF) and increased stem mass fraction (SMF). In contrast, the interstock decreased SMF and increased LMF and RMF. The VL/SAO/SO trees had the highest RMF and proportionally higher Cl- and Na+ allocations in roots than the other plant types. In saline conditions, reductions in leaf ion transport rate and dilution of imported ions by foliar growth nearly halved ion accumulations in leaves of VL/SAO/SO trees. PMID- 14532012 TI - Wood properties of Scots pines (Pinus sylvestris) grown at elevated temperature and carbon dioxide concentration. AB - Impacts of elevated temperature and carbon dioxide concentration ([CO2]) on wood properties of 15-year-old Scots pines (Pinus sylvestris L.) grown under conditions of low nitrogen supply were investigated in open-top chambers. The treatments consisted of (i) ambient temperature and ambient [CO2] (AT+AC), (ii) ambient temperature and elevated [CO2] (AT+EC), (iii) elevated temperature and ambient [CO2] (ET+AC) and (iv) elevated temperature and elevated [CO2] (ET+EC). Wood properties analyzed for the years 1992-1994 included ring width, early- and latewood width and their proportions, intra-ring wood density (minimum, maximum and mean, as well as early- and latewood densities), mean fiber length and chemical composition of the wood (cellulose, hemicellulose, lignin and acetone extractive concentration). Absolute radial growth over the 3-year period was 54% greater in AT+EC trees and 30 and 25% greater in ET+AC and ET+EC trees, respectively, than in AT+AC trees. Neither elevated temperature nor elevated [CO2] had a statistically significant effect on ring width, early- and latewood widths or their proportions. Both latewood density and maximum intra-ring density were increased by elevated [CO2], whereas fiber length was increased by elevated temperature. Hemicellulose concentration decreased and lignin concentration increased significantly in response to elevated temperature. There were no statistically significant interaction effects of elevated temperature and elevated [CO2] on the wood properties, except on earlywood density. PMID- 14532013 TI - Fruit load and canopy shading affect leaf characteristics and net gas exchange of 'Spring' navel orange trees. AB - Five-year-old 'Spring' navel (Citrus sinensis (L.) Osbeck) orange trees were completely defruited, 50% defruited or left fully laden to study effects of fruit load on concentrations of nitrogen (N) and carbohydrate, net assimilation of CO2 (Ac) and stomatal conductance (gs) of mature leaves on clear winter days just before fruit harvest. Leaves on defruited trees were larger, had higher starch concentrations and greater leaf dry mass per area (LDMa) than leaves on fruited trees. Both Ac and gs were more than 40% lower in sunlit leaves on defruited trees than in sunlit leaves on trees with fruit. Leaves immediately adjacent to fruit were smaller, had lower leaf nitrogen and carbohydrate concentrations, lower LDMa and lower Ac than leaves on non-fruiting branches of the same trees. Removing half the crop increased individual fruit mass, but reduced fruit color development. Half the trees were shaded with 50% shade cloth for 4 months before harvest to determine the effects of lower leaf temperature (Tl) and leaf-to-air vapor pressure difference on leaf responses. On relatively warm days when sunlit Tl > 25 degrees C, shade increased Ac and gs, but had no effect on the ratio of internal to ambient CO2 (Ci/Ca) concentration in leaves, implying that high mesophyll temperatures in sunlit leaves were more important than gs in limiting Ac. Sunlit leaves were more photoinhibited than shaded leaves on cooler days when Tl < 25 degrees C. Shade decreased total soluble sugar concentrations in leaves, but had no effect on leaf starch concentrations. Shading had no effects on canopy volume, yield or fruit size, but shaded fruit developed better external color than sun-exposed fruit. Overall, the presence of a normal fruit crop resulted in lower foliar carbohydrate concentrations and higher Ac compared with defruited trees, except on warm days when Ac was reduced by high leaf temperatures. PMID- 14532014 TI - Tracheid diameter is the key trait determining the extent of freezing-induced embolism in conifers. AB - We tested the hypotheses that freezing-induced embolism is related to conduit diameter, and that conifers and angiosperms with conduits of equivalent diameter will exhibit similar losses of hydraulic conductivity in response to freezing. We surveyed the freeze-thaw response of conifers with a broad range of tracheid diameters by subjecting wood segments (root, stem and trunk wood) to a freeze thaw cycle at -0.5 MPa in a centrifuge. Embolism increased as mean tracheid diameter exceeded 30 microm. Tracheids with a critical diameter greater than 43 microm were calculated to embolize in response to freezing and thawing at a xylem pressure of -0.5 MPa. To confirm that freezing-induced embolism is a function of conduit air content, we air-saturated stems of Abies lasiocarpa (Hook.) Nutt. (mean conduit diameter 13.7 +/- 0.7 microm) by pressurizing them 1 to 60 times above atmospheric pressure, prior to freezing and thawing. The air saturation method simulated the effect of increased tracheid size because the degree of super-saturation is proportional to a tracheid volume holding an equivalent amount of dissolved air at ambient pressure. Embolism increased when the dissolved air content was equivalent to a mean tracheid diameter of 30 microm at ambient air pressure. Our centrifuge and air-saturation data show that conifers are as vulnerable to freeze-thaw embolism as angiosperms with equal conduit diameter. We suggest that the hydraulic conductivity of conifer wood is maximized by increasing tracheid diameters in locations where freezing is rare. Conversely, the narrowing of tracheid diameters protects against freezing-induced embolism in cold climates. PMID- 14532015 TI - Root turnover and root necromass accumulation of Norway spruce (Picea abies) are affected by soil acidity. AB - Fine root distribution and turnover were investigated in ca. 40-year-old pure Norway spruce (Picea abies Karst.) stands in Germany, growing on four sites that differed in soil acidity (Ebergotzen < Barbis < Fichtelgebirge = Harz). The density of fine root biomass and necromass in different soil horizons differed among the sites. At one of the most acidic sites (Harz), fine root density in the humus layer was more than twice that at the least acidic site (Ebergotzen). At the two most acidic sites, Fichtelgebirge and Harz, the ratio of biomass to necromass was significantly lower than at Ebergotzen and Barbis, particularly in the subsoil layer. In each stand, clear vertical gradients in fine root length density and root tip density were observed. Most of the roots and the root tips were in the humus layer and in the first mineral soil horizon (0-10 cm). There was a significantly different decrease in specific root length (cm gDM (-1)) and specific root tip density (root tips gDM (-1)) in the more acidified stands Fichtelgebirge and Harz compared with Ebergotzen and Barbis. Fine root production estimated by ingrowth cores and a net method was approximately twice as high in the more acidic stands Fichtelgebirge and Harz compared with Ebergotzen and Barbis. Rates of living fine root biomass turnover were higher at the Fichtelgebirge and Harz sites than at the Ebergotzen site. Rates of necromass turnover were similar at all sites. The results suggest that the accumulation of necromass was not due to a slower disappearance at the more acid sites, but to earlier root death. Roots contributed 46% to root + needle litter and 32% to root + total aboveground litter at the Harz site in 1997. PMID- 14532017 TI - High autumn temperature delays spring bud burst in boreal trees, counterbalancing the effect of climatic warming. AB - The effect of temperature during short-day (SD) dormancy induction was examined in three boreal tree species in a controlled environment. Saplings of Betula pendula Roth, B. pubescens Ehrh. and Alnus glutinosa (L.) Moench. were exposed to 5 weeks of 10-h SD induction at 9, 15 and 21 degrees C followed by chilling at 5 degrees C for 40, 70, 100 and 130 days and subsequent forcing at 15 degrees C in a 24-h photoperiod for 60 days. In all species and with all chilling periods, high temperature during SD dormancy induction significantly delayed bud burst during subsequent flushing at 15 degrees C. In A. glutinosa, high temperature during SD dormancy induction also significantly increased the chilling requirement for dormancy release. Field experiments at 60 degrees N with a range of latitudinal birch populations revealed a highly significant correlation between autumn temperature and days to bud burst in the subsequent spring. September temperature alone explained 20% of the variation between years in time of bud burst. In birch populations from 69 and 71 degrees N, which ceased growing and shed their leaves in August when the mean temperature was 15 degrees C, bud burst occurred later than expected compared with lower latitude populations (56 degrees N) in which dormancy induction took place more than 2 months later at a mean temperature of about 6 degrees C. It is concluded that this autumn temperature response may be important for counterbalancing the potentially adverse effects of higher winter temperatures on dormancy stability of boreal trees during climate warming. PMID- 14532016 TI - Variation in morphological and biochemical O3 injury attributes of mature Jeffrey pine within canopies and between microsites. AB - Crown morphology and leaf tissue chemical and biochemical attributes associated with ozone (O3) injury were assessed in the lower, mid- and upper canopy of Jeffrey pine (Pinus jeffreyi Grev. & Balf.) growing in mesic and xeric microsites in Sequoia National Park, California. Microsites were designated mesic or xeric based on topography and bole growth in response to years of above-average precipitation. In mesic microsites, canopy response to O3 was characterized by thinner branches, earlier needle fall, less chlorotic leaf mottling, and lower foliar antioxidant capacity, especially of the aqueous fraction. In xeric microsites, canopy response to O3 was characterized by higher chlorotic leaf mottling, shorter needles, lower needle chlorophyll concentration, and greater foliar antioxidant capacity. Increased leaf chlorotic mottle in xeric microsites was related to drought stress and increased concurrent internal production of highly reactive oxygen species, and not necessarily to stomatal O3 uptake. Within canopy position also influenced the expression of O3 injury in Jeffrey pine. PMID- 14532018 TI - Potential role for a novel AP180-related protein during endocytosis in MDCK cells. AB - Clathrin assembly protein, AP180, was originally identified as a brain-specific protein localized to the presynaptic junction. AP180 acts to limit vesicle size and maintain a pool of releasable synaptic vesicles during rapid recycling. In this study, we show that polarized epithelial Madin-Darby canine kidney (MDCK) cells express two AP180-related proteins: the ubiquitously expressed 62-kDa clathrin assembly lymphoid myeloid leukemia (CALM, AP180-2) protein and a novel high-molecular-weight homolog that we have named AP180-3. Sequence analysis of AP180-3 expressed in MDCK cells shows high homology to AP180 from rat brain. AP180-3 contains conserved motifs found in brain-specific AP180, including the epsin NH2-terminal homology (ENTH) domain, the binding site for the alpha-subunit of AP-2, and DLL repeats. Our studies show that AP180-3 from MDCK cells forms complexes with AP-2 and clathrin and that membrane recruitment of these complexes is modulated by phosphorylation. We demonstrate by immunohistochemistry that AP180-3 is localized to cytoplasmic vesicles in MDCK cells and is also present in tubule epithelial cells from mouse kidney. We observed by immunodetection that a high-molecular-weight AP180-related protein is expressed in numerous cells in addition to MDCK cells. PMID- 14532019 TI - Rho GTPase signaling modulates cell shape and contractile phenotype in an isoactin-specific manner. AB - Rho family small GTPases (Rho, Rac, and Cdc42) play an important role in cell motility, adhesion, and cell division by signaling reorganization of the actin cytoskeleton. Here, we report an isoactin-specific, Rho GTPase-dependent signaling cascade in cells simultaneously expressing smooth muscle and nonmuscle actin isoforms. We transfected primary cultures of microvascular pericytes, cells related to vascular smooth muscle cells, with various Rho-related and Rho specific expression plasmids. Overexpression of dominant positive Rho resulted in the formation of nonmuscle actin-containing stress fibers. At the same time, alpha-vascular smooth muscle actin (alphaVSMactin) containing stress fibers were disassembled, resulting in a dramatic reduction in cell size. Rho activation also yielded a disassembly of smooth muscle myosin and nonmuscle myosin from stress fibers. Overexpression of wild-type Rho had similar but less dramatic effects. In contrast, dominant negative Rho and C3 exotransferase or dominant positive Rac and Cdc42 expression failed to alter the actin cytoskeleton in an isoform specific manner. The loss of smooth muscle contractile protein isoforms in pericyte stress fibers, together with a concomitant decrease in cell size, suggests that Rho activation influences "contractile" phenotype in an isoactin specific manner. This, in turn, should yield significant alteration in microvascular remodeling during developmental and pathologic angiogenesis. PMID- 14532020 TI - Glutathione protects Lactococcus lactis against oxidative stress. AB - Glutathione was found in several dairy Lactococcus lactis strains grown in M17 medium. None of these strains was able to synthesize glutathione. In chemically defined medium, L. lactis subsp. cremoris strain SK11 was able to accumulate up to approximately 60 mM glutathione when this compound was added to the medium. Stationary-phase cells of strain SK11 grown in chemically defined medium supplemented with glutathione showed significantly increased resistance (up to fivefold increased resistance) to treatment with H2O2 compared to the resistance of cells without intracellular glutathione. The resistance to H2O2 treatment was found to be dependent on the accumulation of glutathione in 16 strains of L. lactis tested. We propose that by taking up glutathione, L. lactis might activate a glutathione-glutathione peroxidase-glutathione reductase system in stationary phase cells, which catalyzes the reduction of H2O2. Glutathione reductase, which reduces oxidized glutathione, was detectable in most strains of L. lactis, but the activities of different strains were very variable. In general, the glutathione reductase activities of L. lactis subsp. lactis are higher than those of L. lactis subsp. cremoris, and the activities were much higher when strains were grown aerobically. In addition, glutathione peroxidase is detectable in strain SK11, and the level was fivefold greater when the organism was grown aerobically than when the organism was grown anaerobically. Therefore, the presence of glutathione in L. lactis could result in greater stability under storage conditions and quicker growth upon inoculation, two important attributes of successful starter cultures. PMID- 14532021 TI - Purification and characterization of a novel bacteriocin produced by Enterococcus faecalis strain RJ-11. AB - Lactic acid bacteria exhibiting activity against the gram-positive bacterium Bacillus subtilis were isolated from rice bran. One of the isolates, identified as Enterococcus faecalis RJ-11, exhibited a wide spectrum of growth inhibition with various gram-positive bacteria. A bacteriocin purified from culture fluid, designated enterocin RJ-11, was heat stable and was not sensitive to acid and alkaline conditions, but it was sensitive to several proteolytic enzymes. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that enterocin RJ-11 had a molecular weight of 5,000 in its monomeric form. The amino acid sequence determined for purified enterocin RJ-11 exhibited high levels of similarity to the sequences of enterocins produced by Enterococcus faecium. PMID- 14532023 TI - A potential food-grade cloning vector for Streptococcus thermophilus that uses cadmium resistance as the selectable marker. AB - A potential food-grade cloning vector, pND919, was constructed and transformed into S. thermophilus ST3-1, a plasmid-free strain. The vector contains DNAs from two different food-approved organisms, Streptococcus thermophilus and Lactococcus lactis. The 5.0-kb pND919 is a derivative of the cloning vector pND918 (9.3 kb) and was constructed by deletion of the 4.3-kb region of pND918 which contained DNA from non-food-approved organisms. pND919 carries a heterologous native cadmium resistance selectable marker from L. lactis M71 and expresses the Cd(r) phenotype in S. thermophilus transformants. With the S. thermophilus replicon derived from the shuttle vector pND913, pND919 is able to replicate in the two S. thermophilus industrial strains tested, ST3-1 and ST4-1. Its relatively high retention rate in S. thermophilus further indicates its usefulness as a potential food-grade cloning vector. To our knowledge, this is the first report of a replicative potential food-grade vector for the industrially important organism S. thermophilus. PMID- 14532022 TI - Diversity of nitrile hydratase and amidase enzyme genes in Rhodococcus erythropolis recovered from geographically distinct habitats. AB - A molecular screening approach was developed in order to amplify the genomic region that codes for the alpha- and beta-subunits of the nitrile hydratase (NHase) enzyme in rhodococci. Specific PCR primers were designed for the NHase genes from a collection of nitrile-degrading actinomycetes, but amplification was successful only with strains identified as Rhodococcus erythropolis. A hydratase PCR product was also obtained from R. erythropolis DSM 43066(T), which did not grow on nitriles. Southern hybridization of other members of the nitrile degrading bacterial collection resulted in no positive signals other than those for the R. erythropolis strains used as positive controls. PCR-restriction fragment length polymorphism-single-strand conformational polymorphism (PRS) analysis of the hydratases in the R. erythropolis strains revealed unique patterns that mostly correlated with distinct geographical sites of origin. Representative NHases were sequenced, and they exhibited more than 92.4% similarity to previously described NHases. The phylogenetic analysis and deduced amino acid sequences suggested that the novel R. erythropolis enzymes belonged to the iron-type NHase family. Some different residues in the translated sequences were located near the residues involved in the stabilization of the NHase active site, suggesting that the substitutions could be responsible for the different enzyme activities and substrate specificities observed previously in this group of actinomycetes. A similar molecular screening analysis of the amidase gene was performed, and a correlation between the PRS patterns and the geographical origins identical to the correlation found for the NHase gene was obtained, suggesting that there was coevolution of the two enzymes in R. erythropolis. Our findings indicate that the NHase and amidase genes present in geographically distinct R. erythropolis strains are not globally mixed. PMID- 14532024 TI - Engineering Escherichia coli for increased productivity of serine-rich proteins based on proteome profiling. AB - Variations in proteome profiles of Escherichia coli in response to the overproduction of human leptin, a serine-rich (11.6% of total amino acids) protein, were examined by two-dimensional gel electrophoresis. The levels of heat shock proteins increased, while those of protein elongation factors, 30S ribosomal protein, and some enzymes involved in amino acid biosynthesis decreased, after leptin overproduction. Most notably, the levels of enzymes involved in the biosynthesis of serine family amino acids significantly decreased. Based on this information, we designed a strategy to enhance the leptin productivity by manipulating the cysK gene, encoding cysteine synthase A. By coexpression of the cysK gene, we were able to increase the cell growth rate by approximately twofold. Also, the specific leptin productivity could be increased by fourfold. In addition, we found that cysK coexpression can improve the production of another serine-rich protein, interleukin-12 beta chain, suggesting that this strategy may be useful for the production of other serine rich proteins as well. The approach taken in this study should be useful in designing a strategy for improving recombinant protein production. PMID- 14532026 TI - Development of an improved selective agar medium for isolation of Yersinia pestis. AB - Existing media designed for selective isolation of clinically important members of the genus Yersinia were found to be unsatisfactory for the growth and isolation of Yersinia pestis. We report the development of a new selective agar medium (termed BIN) that supports the growth of Y. pestis. The development of the formulation of this medium was based on a fluorescence screening system designed for monitoring bacterial growth on semisolid media, using a green fluorescent protein-expressing strain. High-throughput combinatorial experiments can be conducted for the quantitative evaluation of the effect of different medium components on growth. Generation of fluorescence plots in this system, using microplates, allowed the quantitative evaluation of the growth rate of Y. pestis EV76 cultures in different agar compositions. The final BIN formulation is based on brain heart infusion agar, to which the selective agents irgasan, cholate salts, crystal violet, and nystatin were introduced. It was found that BIN agar is more efficient in supporting colony formation and recovery of Y. pestis than are the conventional semisolid media MacConkey agar and Yersinia-selective agar (cefsulodin-irgasan-novobiocin agar). The advantage of BIN over other media has been also demonstrated in recovering virulent Y. pestis from the mixed bacterial populations found in decaying carcasses of infected mice. The BIN medium is suggested as a selective medium for isolation and recovery of Y. pestis from various backgrounds. PMID- 14532025 TI - Detection, quantitation, and phylogenetic analysis of noroviruses in Japanese oysters. AB - Noroviruses (NVs) cause many cases of oyster- or clam-associated gastroenteritis in various countries. We collected 191 samples from Japanese oysters intended for raw consumption that had been harvested from the sea in two different areas between December 2001 and February 2002. To detect, quantitate, and phylogenetically analyze the NV genome in purified concentrates from the stomachs and digestive diverticula of these oysters, we amplified the NV capsid gene by reverse transcription-PCR. Phylogenetic analysis was performed by using the neighbor-joining method. We detected the NV genome in 17 of 191 oysters (9%). Phylogenetic analysis indicated genogroup I (Norwalk virus type) in 3 of the 17 oysters and genogroup II (Snow Mountain virus type) in the other 14. Both genogroups showed wide genetic diversity. To quantify the NV capsid gene in these oysters, we performed real-time PCR using genogroup-specific probes. More than 10(2) copies of the NV genome were detected in 11 of 17 oysters. The results suggested that about 10% of Japanese oysters intended for raw consumption harbored NVs, and more than 50% of those oysters in which NVs were detected had a large amount. PMID- 14532027 TI - Conditional survival as a selection strategy to identify plant-inducible genes of Pseudomonas syringae. AB - A novel strategy termed habitat-inducible rescue of survival (HIRS) was developed to identify genes of Pseudomonas syringae that are induced during growth on bean leaves. This strategy is based on the complementation of metXW, two cotranscribed genes that are necessary for methionine biosynthesis and required for survival of P. syringae on bean leaves exposed to conditions of low humidity. We constructed a promoter trap vector, pTrap, containing a promoterless version of the wild-type P. syringae metXW genes. Only with an active promoter fused to metXW on pTrap did this plasmid restore methionine prototrophy to the P. syringae metXW mutant B7MX89 and survival of this strain on bean leaves. To test this method, a partial library of P. syringae genomic DNA was constructed in pTrap and a total of 1,400 B7MX89 pTrap clones were subjected to HIRS selection on bean leaves. This resulted in the enrichment of five clones, each with a unique RsaI restriction pattern of their DNA insert. Sequence analysis of these clones revealed those P. syringae genes for which putative plant-inducible activity could be assigned. Promoter activity experiments with a gfp reporter gene revealed that these plant inducible gene promoters had very low levels of expression in minimal medium. Based on green fluorescent protein fluorescence levels, it appears that many P. syringae genes have relatively low expression levels and that the metXW HIRS strategy is a sensitive method to detect weakly expressed P. syringae genes that are active on plants. Furthermore, we found that protected sites on the leaf surface provided a higher level of enrichment for P. syringae expressing metXW than exposed sites. Thus, the metXW HIRS strategy should lead to the identification of P. syringae genes that are expressed primarily in these areas on the leaf. PMID- 14532028 TI - Identification and genetic characterization of a novel proteinase, PrtR, from the human isolate Lactobacillus rhamnosus BGT10. AB - A novel proteinase, PrtR, produced by the human vaginal isolate Lactobacillus rhamnosus strain BGT10 was identified and genetically characterized. The prtR gene and flanking regions were cloned and sequenced. The deduced amino acid sequence of PrtR shares characteristics that are common for other cell envelope proteinases (CEPs) characterized to date, but in contrast to the other cell surface subtilisin-like serine proteinases, it has a smaller and somewhat different B domain and lacks the helix domain, and the anchor domain has a rare sorting signal sequence. Furthermore, PrtR lacks the insert domain, which otherwise is situated inside the catalytic serine protease domain of all CEPs, and has a different cell wall spacer (W) domain similar to that of the cell surface antigen I and II polypeptides expressed by oral and vaginal streptococci. Moreover, the PrtR W domain exhibits significant sequence homology to the consensus sequence that has been shown to be the hallmark of human intestinal mucin protein. According to its alpha(S1)- and beta-casein cleavage efficacy, PrtR is an efficient proteinase at pH 6.5 and is distributed throughout all L. rhamnosus strains tested. Proteinase extracts of the BGT10 strain obtained with Ca(2+)-free buffer at pH 6.5 were proteolytically active. The prtR promoter-like sequence was determined, and the minimal promoter region was defined by use of prtR-gusA operon fusions. The prtR expression is Casitone dependent, emphasizing that nitrogen depletion elevates its transcription. This is in correlation with the catalytic activity of the PrtR proteinase. PMID- 14532029 TI - Establishment of an arbitrary PCR for rapid identification of Tn917 insertion sites in Staphylococcus epidermidis: characterization of biofilm-negative and nonmucoid mutants. AB - Transposon mutagenesis with the Enterococcus faecalis transposon Tn917 is a genetic approach frequently used to identify genes related with specific phenotypes in gram-positive bacteria. We established an arbitrary PCR for the rapid and easy identification of Tn917 insertion sites in Staphylococcus epidermidis with six independent, well-characterized biofilm-negative Tn917 transposon mutants, which were clustered in the icaADBC gene locus or harbor Tn917 in the regulatory gene rsbU. For all six of these mutants, short chromosomal DNA fragments flanking both transposon ends could be amplified. All fragments were sufficient to correctly identify the Tn917 insertion sites in the published S. epidermidis genomes. By using this technique, the Tn917 insertion sites of three not-yet-characterized biofilm-negative or nonmucoid mutants were identified. In the biofilm-negative and nonmucoid mutant M12, Tn917 is inserted into a gene homologous to the regulatory gene purR of Bacillus subtilis and Staphylococcus aureus. The Tn917 insertions of the nonmucoid but biofilm-positive mutants M16 and M20 are located in genes homologous to components of the phosphoenolpyruvate-sugar phosphotransferase system (PTS) of B. subtilis, S. aureus, and Staphylococcus carnosus, indicating an influence of the PTS on the mucoid phenotype in S. epidermidis. PMID- 14532030 TI - Development of a PCR-enzyme immunoassay oligoprobe detection method for Toxoplasma gondii oocysts, incorporating PCR controls. AB - Infections caused by Toxoplasma gondii are widely prevalent in animals and humans throughout the world. In the United States, an estimated 23% of adolescents and adults have laboratory evidence of T. gondii infection. T. gondii has been identified as a major opportunistic pathogen in immunocompromised individuals, in whom it can cause life-threatening disease. Water contaminated with feces from domestic cats or other felids may be an important source of human exposure to T. gondii oocysts. Because of the lack of information regarding the prevalence of T. gondii in surface waters, there is a clear need for a rapid, sensitive method to detect T. gondii from water. Currently available animal models and cell culture methods are time-consuming, expensive, and labor-intensive, requiring days or weeks for results to be obtained. Detection of T. gondii nucleic acid by PCR has become the preferred method. We have developed a PCR amplification and detection method for T. gondii oocyst nucleic acid that incorporates the use of hot-start amplification to reduce nonspecific primer annealing, uracil-N-glycosylase to prevent false-positive results due to carryover contamination, an internal standard control to identify false-negative results due to inadequate removal of sample inhibition, and PCR product oligoprobe confirmation using a nonradioactive DNA hybridization immunoassay. This method can provide positive, confirmed results in less than 1 day. Fewer than 50 oocysts can be detected following recovery of oocyst DNA. Development of a T. gondii oocyst PCR detection method will provide a useful technique to estimate the levels of T. gondii oocysts present in surface waters. PMID- 14532031 TI - Membrane fluidity adjustments in ethanol-stressed Oenococcus oeni cells. AB - The effect of ethanol on the cytoplasmic membrane of Oenococcus oeni cells and the role of membrane changes in the acquired tolerance to ethanol were investigated. Membrane tolerance to ethanol was defined as the resistance to ethanol-induced leakage of preloaded carboxyfluorescein (cF) from cells. To probe the fluidity of the cytoplasmic membrane, intact cells were labeled with doxyl stearic acids and analyzed by electron spin resonance spectroscopy. Although the effect of ethanol was noticeable across the width of the membrane, we focused on fluidity changes at the lipid-water interface. Fluidity increased with increasing concentrations of ethanol. Cells responded to growth in the presence of 8% (vol/vol) ethanol by decreasing fluidity. Upon exposure to a range of ethanol concentrations, these adapted cells had reduced fluidity and cF leakage compared with cells grown in the absence of ethanol. Analysis of the membrane composition revealed an increase in the degree of fatty acid unsaturation and a decrease in the total amount of lipids in the cells grown in the presence of 8% (vol/vol) ethanol. Preexposure for 2 h to 12% (vol/vol) ethanol also reduced membrane fluidity and cF leakage. This short-term adaptation was not prevented in the presence of chloramphenicol, suggesting that de novo protein synthesis was not involved. We found a strong correlation between fluidity and cF leakage for all treatments and alcohol concentrations tested. We propose that the protective effect of growth in the presence of ethanol is, to a large extent, based on modification of the physicochemical state of the membrane, i.e., cells adjust their membrane permeability by decreasing fluidity at the lipid-water interface. PMID- 14532032 TI - Substrate preferences in biodesulfurization of diesel range fuels by Rhodococcus sp. strain ECRD-1. AB - The range of sulfur compounds in fuel oil and the substrate range and preference of the biocatalytic system determine the maximum extent to which sulfur can be removed by biodesulfurization. We show that the biodesulfurization apparatus in Rhodococcus sp. strain ECRD-1 is able to attack all isomers of dibenzothiophene including those with at least four pendant carbons, with a slight preference for those substituted in the alpha-position. With somewhat less avidity, this apparatus is also able to attack substituted benzothiophenes with between two and seven pendant carbons. Some compounds containing sulfidic sulfur are also susceptible to desulfurization, although we have not yet been able to determine their molecular identities. PMID- 14532033 TI - Functional analysis of the gene cluster involved in production of the bacteriocin circularin A by Clostridium beijerinckii ATCC 25752. AB - A region of 12 kb flanking the structural gene of the cyclic antibacterial peptide circularin A of Clostridium beijerinckii ATCC 25752 was sequenced, and the putative proteins involved in the production and secretion of circularin A were identified. The genes are tightly organized in overlapping open reading frames. Heterologous expression of circularin A in Enterococcus faecalis was achieved, and five genes were identified as minimally required for bacteriocin production and secretion. Two of the putative proteins, CirB and CirC, are predicted to contain membrane-spanning domains, while CirD contains a highly conserved ATP-binding domain. Together with CirB and CirC, this ATP-binding protein is involved in the production of circularin A. The fifth gene, cirE, confers immunity towards circularin A when expressed in either Lactococcus lactis or E. faecalis and is needed in order to allow the bacteria to produce bacteriocin. Additional resistance against circularin A is conferred by the activity of the putative transporter consisting of CirB and CirD. PMID- 14532034 TI - Anaerobic degradation of flavonoids by Clostridium orbiscindens. AB - An anaerobic, quercetin-degrading bacterium was isolated from human feces and identified as Clostridium orbiscindens by comparative 16S rRNA gene sequence analysis. The organism was tested for its ability to transform several flavonoids. The isolated C. orbiscindens strain converted quercetin and taxifolin to 3,4-dihydroxyphenylacetic acid; luteolin and eriodictyol to 3-(3,4 dihydroxyphenyl)propionic acid; and apigenin, naringenin, and phloretin to 3-(4 hydroxyphenyl)propionic acid, respectively. Genistein and daidzein were not utilized. The glycosidic bonds of luteolin-3-glucoside, luteolin-5-glucoside, naringenin-7-neohesperidoside (naringin), quercetin-3-glucoside, quercetin-3 rutinoside (rutin), and phloretin-2'-glucoside were not cleaved. Based on the intermediates and products detected, pathways for the degradation of the flavonol quercetin and the flavones apigenin and luteolin are proposed. To investigate the numerical importance of C. orbiscindens in the human intestinal tract, a species specific oligonucleotide probe was designed and tested for its specificity. Application of the probe to fecal samples from 10 human subjects proved the presence of C. orbiscindens in 8 out of the 10 samples tested. The numbers ranged from 1.87 x 10(8) to 2.50 x 10(9) cells g of fecal dry mass(-1), corresponding to a mean count of 4.40 x 10(8) cells g of dry feces(-1). PMID- 14532035 TI - Peptide surface display and secretion using two LPXTG-containing surface proteins from Lactobacillus fermentum BR11. AB - A locus encoding two repetitive proteins that have LPXTG cell wall anchoring signals from Lactobacillus fermentum BR11 has been identified by using an antiserum raised against whole L. fermentum BR11 cells. The first protein, Rlp, is similar to the Rib surface protein from Streptococcus agalactiae, while the other protein, Mlp, is similar to the mucus binding protein Mub from Lactobacillus reuteri. It was shown that multiple copies of mlp exist in the genome of L. fermentum BR11. Regions of Rlp, Mlp, and the previously characterized surface protein BspA were used to surface display or secrete heterologous peptides in L. fermentum. The peptides tested were 10 amino acids of the human cystic fibrosis transmembrane regulator protein and a six-histidine epitope (His(6)). The BspA promoter and secretion signal were used in combination with the Rlp cell wall sorting signal to express, export, and covalently anchor the heterologous peptides to the cell wall. Detection of the cell surface protein fusions revealed that Rlp was a significantly better surface display vector than BspA despite having lower cellular levels (0.7 mg per liter for the Rlp fusion compared with 4 mg per liter for the BspA fusion). The mlp promoter and encoded secretion signal were used to express and export large (328-kDa at 10 mg per liter) and small (27-kDa at 0.06 mg per liter) amino-terminal fragments of the Mlp protein fused to the His(6) and CFTR peptides or His(6) peptide, respectively. Therefore, these newly described proteins from L. fermentum BR11 have potential as protein production and targeting vectors. PMID- 14532037 TI - Identification of Naegleria fowleri in domestic water sources by nested PCR. AB - The free-living amoeboflagellate Naegleria fowleri is the causative agent of primary amoebic meningoencephalitis (PAM), a rapidly fatal disease of the central nervous system. In the United States, the disease is generally acquired while swimming and diving in freshwater lakes and ponds. In addition to swimming, exposure to N. fowleri and the associated disease can occur by total submersion in bathwater or small backyard wading pools. In the present study, swipe samples and residual pipe water from homes in Arizona were examined for N. fowleri by nested PCR due to the death of two previously healthy children from PAM. Since neither child had a history of swimming in a freshwater lake or pond prior to the onset of disease symptoms, the domestic water supply was the suspected source of infection. Of 19 samples collected from bathroom and kitchen pipes and sink traps, 17 samples were positive for N. fowleri by PCR. A sample from a Micro-Wynd II filter was obtained by passing water from bathtubs through the filter. Organisms attached to the filter also tested positive by PCR. The two samples that tested negative for N. fowleri were one that was obtained from a kitchen sink trap and a swipe sample from the garbage disposal of one home. PMID- 14532038 TI - Mycotoxin fumonisin B1 increases intestinal colonization by pathogenic Escherichia coli in pigs. AB - Fumonisin B(1) (FB(1)) is a mycotoxin that commonly occurs in maize. FB(1) causes a variety of toxic effects in different animal species and has been implicated as a contributing factor of esophageal cancers in humans. In the present study, we examined the effect of dietary exposure to FB(1) on intestinal colonization by pathogenic Escherichia coli associated with extraintestinal infection. Three-week old weaned pigs were given FB(1) by gavage as a crude extract or as a purified toxin at a dose of 0.5 mg/kg of body weight daily for 6 days. On the last day of the toxin treatment, the pigs were orally inoculated with an extraintestinal pathogenic E. coli strain. All animals were euthanized 24 h later, necropsies were performed, and tissues were taken for bacterial counts and light microscopic examination. Ingestion of FB(1) had only a minimal effect on animal weight gain, did not cause any macroscopic or microscopic lesions, and did not change the plasma biochemical profile. However, colonization of the small and large intestines by an extraintestinal pathogenic E. coli strain was significantly increased. Our results show that FB(1) is a predisposing factor to infectious disease and that the pig can be used as a model for the study of the consequences of ingesting mycotoxin-contaminated food. PMID- 14532039 TI - Rapid screening for freshwater bacterial groups by using reverse line blot hybridization. AB - The identification of phylogenetic clusters of bacteria that are common in freshwater has provided a basis for probe design to target important freshwater groups. We present a set of 16S ribosomal RNA gene-based oligonucleotide probes specific for 15 of these freshwater clusters. The probes were applied in reverse line blot hybridization, a simple method that enables the rapid screening of PCR products from many samples against an array of probes. The optimized assay was made stringent to discriminate at approximately the single-mismatch level. This made 10 of the probes highly specific, with at least two mismatches to the closest noncluster member in the global database. Screening of PCR products from bacterioplankton of 81 diverse lakes from Belgium, The Netherlands, Denmark, Sweden, and Norway showed that the respective probes were reactive against 5 to 100% of the lake samples. Positive reactivity of six highly specific probes showed that bacteria from actinobacterial clusters ACK-M1 and Sta2-30 and from verrucomicrobial cluster CLO-14 occurred in at least 90% of the investigated lakes. Furthermore, bacteria from alpha-proteobacterial cluster LD12 (closely related to the marine SAR11 cluster), beta-proteobacterial cluster LD28 and cyanobacterial cluster Synechococcus 6b occurred in more than 70% of the lakes. Reverse line blot hybridization is a new tool in microbial ecology that will facilitate research on distribution and habitat specificity of target species at relatively low costs. PMID- 14532040 TI - Stimulating the in situ activity of Geobacter species to remove uranium from the groundwater of a uranium-contaminated aquifer. AB - The potential for removing uranium from contaminated groundwater by stimulating the in situ activity of dissimilatory metal-reducing microorganisms was evaluated in a uranium-contaminated aquifer located in Rifle, Colo. Acetate (1 to 3 mM) was injected into the subsurface over a 3-month period via an injection gallery composed of 20 injection wells, which was installed upgradient from a series of 15 monitoring wells. U(VI) concentrations decreased in as little as 9 days after acetate injection was initiated, and within 50 days uranium had declined below the prescribed treatment level of 0.18 micro M in some of the monitoring wells. Analysis of 16S ribosomal DNA (rDNA) sequences and phospholipid fatty acid profiles demonstrated that the initial loss of uranium from the groundwater was associated with an enrichment of Geobacter species in the treatment zone. Fe(II) in the groundwater also increased during this period, suggesting that U(VI) reduction was coincident with Fe(III) reduction. As the acetate injection continued over 50 days there was a loss of sulfate from the groundwater and an accumulation of sulfide and the composition of the microbial community changed. Organisms with 16S rDNA sequences most closely related to those of sulfate reducers became predominant, and Geobacter species became a minor component of the community. This apparent switch from Fe(III) reduction to sulfate reduction as the terminal electron accepting process for the oxidation of the injected acetate was associated with an increase in uranium concentration in the groundwater. These results demonstrate that in situ bioremediation of uranium contaminated groundwater is feasible but suggest that the strategy should be optimized to better maintain long-term activity of Geobacter species. PMID- 14532041 TI - Engineering redox cofactor regeneration for improved pentose fermentation in Saccharomyces cerevisiae. AB - Pentose fermentation to ethanol with recombinant Saccharomyces cerevisiae is slow and has a low yield. A likely reason for this is that the catabolism of the pentoses D-xylose and L-arabinose through the corresponding fungal pathways creates an imbalance of redox cofactors. The process, although redox neutral, requires NADPH and NAD+, which have to be regenerated in separate processes. NADPH is normally generated through the oxidative part of the pentose phosphate pathway by the action of glucose-6-phosphate dehydrogenase (ZWF1). To facilitate NADPH regeneration, we expressed the recently discovered gene GDP1, which codes for a fungal NADP+-dependent D-glyceraldehyde-3-phosphate dehydrogenase (NADP GAPDH) (EC 1.2.1.13), in an S. cerevisiae strain with the D-xylose pathway. NADPH regeneration through an NADP-GAPDH is not linked to CO2 production. The resulting strain fermented D-xylose to ethanol with a higher rate and yield than the corresponding strain without GDP1; i.e., the levels of the unwanted side products xylitol and CO2 were lowered. The oxidative part of the pentose phosphate pathway is the main natural path for NADPH regeneration. However, use of this pathway causes wasteful CO2 production and creates a redox imbalance on the path of anaerobic pentose fermentation to ethanol because it does not regenerate NAD+. The deletion of the gene ZWF1 (which codes for glucose-6-phosphate dehydrogenase), in combination with overexpression of GDP1 further stimulated D xylose fermentation with respect to rate and yield. Through genetic engineering of the redox reactions, the yeast strain was converted from a strain that produced mainly xylitol and CO2 from D-xylose to a strain that produced mainly ethanol under anaerobic conditions. PMID- 14532042 TI - Dual resistance to Bacillus thuringiensis Cry1Ac and Cry2Aa toxins in Heliothis virescens suggests multiple mechanisms of resistance. AB - One strategy for delaying evolution of resistance to Bacillus thuringiensis crystal (Cry) endotoxins is the production of multiple Cry toxins in each transgenic plant (gene stacking). This strategy relies upon the assumption that simultaneous evolution of resistance to toxins that have different modes of action will be difficult for insect pests. In B. thuringiensis-transgenic (Bt) cotton, production of both Cry1Ac and Cry2Ab has been proposed to delay resistance of Heliothis virescens (tobacco budworm). After previous laboratory selection with Cry1Ac, H. virescens strains CXC and KCBhyb developed high levels of cross-resistance not only to toxins similar to Cry1Ac but also to Cry2Aa. We studied the role of toxin binding alteration in resistance and cross-resistance with the CXC and KCBhyb strains. In toxin binding experiments, Cry1A and Cry2Aa toxins bound to brush border membrane vesicles from CXC, but binding of Cry1Aa was reduced for the KCBhyb strain compared to susceptible insects. Since Cry1Aa and Cry2Aa do not share binding proteins in H. virescens, our results suggest occurrence of at least two mechanisms of resistance in KCBhyb insects, one of them related to reduction of Cry1Aa toxin binding. Cry1Ac bound irreversibly to brush border membrane vesicles (BBMV) from YDK, CXC, and KCBhyb larvae, suggesting that Cry1Ac insertion was unaffected. These results highlight the genetic potential of H. virescens to become resistant to distinct Cry toxins simultaneously and may question the effectiveness of gene stacking in delaying evolution of resistance. PMID- 14532043 TI - Effects of iron limitation on adherence and cell surface carbohydrates of Corynebacterium diphtheriae strains. AB - Iron limitation may cause bacterial pathogens to grow more slowly; however, it may also stimulate these microorganisms to produce greater tissue damage, given that many virulence factors are controlled by the iron supply in the environment. The present study investigated the influence of low iron availability on the expression of proteins and surface sugar residues of two toxigenic strains of Corynebacterium diphtheriae subsp. mitis and evaluated their adherence to human group B erythrocytes and HEp-2 cells. A comparison was made between bacteria grown in (i) Trypticase soy broth (TSB), (ii) TSB treated with dipyridyl to deplete free iron, and (iii) TSB enriched with FeCl(3). The effects of iron concentration on adhesive properties were different for strains 241 and CDC E8392, of the sucrose-fermenting and non-sucrose-fermenting biotypes, respectively. Iron-limited conditions enhanced interaction of strain 241 with erythrocytes and HEp-2 cells. Inhibition assays suggested the involvement of nonfimbrial protein combination 67-72p on hemagglutination of diphtheria bacilli grown under iron-limited conditions. Conversely, iron limitation inhibited adherence to glass and expression of electron-dense material on the bacterial surface. Lectin binding assays demonstrated a reduction in the number of sialic acid residues and an increase in D-mannose and D-galactose residues on the surfaces of both strains. Thus, iron exerts a regulatory role on adhesive properties of diphtheria bacilli, and low iron availability modulates the expression of C. diphtheriae surface carbohydrate moieties. The significant changes in the degree of lectin binding specific for D-mannose, D-galactose and sialic acid residues may have an effect on binding of host cells. The expression of dissimilar microbial virulence determinants may be coordinately controlled by common regulatory systems. For C. diphtheriae, the present results imply regulation of adherence and slime production as part of a global response to iron limited environmental conditions that includes derepression of genes for the synthesis of cytotoxin and siderophores and for transport of the Fe(III) siderophore complexes. PMID- 14532044 TI - PCR detection and analysis of the free-living amoeba Naegleria in hot springs in Yellowstone and Grand Teton National Parks. AB - Free-living thermotolerant amoebae pose a significant health risk to people who soak and swim in habitats suitable for their growth, such as hot springs. In this survey of 23 different hot springs in Yellowstone and Grand Teton National Parks, we used PCR with primer sets specific for Naegleria to detect three sequence types that represent species not previously described, as well as a fourth sequence type identified as the pathogen Naegleria fowleri. PMID- 14532045 TI - Role of ectoine in Vibrio cholerae osmoadaptation. AB - Vibrio cholerae is both an intestinal pathogen and a microbe in the estuarine community. To persist in the estuarine environment, V. cholerae must adjust to changes in ionic composition and osmolarity. These changes in the aquatic environment have been correlated with cholera epidemics. In this work, we study the response of V. cholerae to increases in environmental osmolarity. Optimal growth of V. cholerae in minimal medium requires supplementation with 200 mM NaCl and KCl. However, when the NaCl concentration is increased beyond 200 mM, a proportionate delay in growth is observed. During this delay in growth, osmotic equilibrium is reached by cytoplasmic accumulation of small, uncharged solutes that are compatible with growth. We show that synthesis of the compatible solute ectoine and transport of the compatible solute glycine betaine impact the length of the osmoadaptive growth delay. We also demonstrate that high-osmolarity adapted V. cholerae displays a growth advantage when competed against unadapted cells in high-osmolarity medium. In contrast, low-osmolarity-adapted V. cholerae displays no growth advantage when competed against high-osmolarity-adapted cells in low-osmolarity medium. These results may have implications for V. cholerae population dynamics when seawater and freshwater and their attendant microbes mix. PMID- 14532046 TI - Population dynamics of Vibrio fischeri during infection of Euprymna scolopes. AB - The luminous bacterium Vibrio fischeri colonizes a specialized light-emitting organ within its squid host, Euprymna scolopes. Newly hatched juvenile squid must acquire their symbiont from ambient seawater, where the bacteria are present at low concentrations. To understand the population dynamics of V. fischeri during colonization more fully, we used mini-Tn7 transposons to mark bacteria with antibiotic resistance so that the growth of their progeny could be monitored. When grown in culture, there was no detectable metabolic burden on V. fischeri cells carrying the transposon, which inserts in single copy in a specific intergenic region of the V. fischeri genome. Strains marked with mini-Tn7 also appeared to be equivalent to the wild type in their ability to infect and multiply within the host during coinoculation experiments. Studies of the early stages of colonization suggested that only a few bacteria became associated with symbiotic tissue when animals were exposed for a discrete period (3 h) to an inoculum of V. fischeri cells equivalent to natural population levels; nevertheless, all these hosts became infected. When three differentially marked strains of V. fischeri were coincubated with juvenile squid, the number of strains recovered from an individual symbiotic organ was directly dependent on the size of the inoculum. Further, these results indicated that, when exposed to low numbers of V. fischeri, the host may become colonized by only one or a few bacterial cells, suggesting that symbiotic infection is highly efficient. PMID- 14532047 TI - Tri16 is required for esterification of position C-8 during trichothecene mycotoxin production by Fusarium sporotrichioides. AB - We previously characterized Tri1, a gene required for hydroxylation of the C-8 position during trichothecene mycotoxin biosynthesis in Fusarium sporotrichioides NRRL 3299. Sequence analysis of the region surrounding Tri1 revealed a gene, named Tri16, which could encode an acyltransferase. Unlike the wild-type parent strain NRRL 3299, which accumulates primarily T-2 toxin along with low levels of diacetoxyscirpenol (DAS) and neosolaniol (NEO) and trace amounts of 8-propionyl neosolaniol (P-NEO) and 8-isobutyryl-neosolaniol (B-NEO), mutants containing a disruption of Tri16 were blocked in the production of the three C-8 esterified compounds T-2 toxin, P-NEO, and B-NEO and accumulated the C-8-hydroxylated compound NEO along with secondary levels of DAS. These data indicate that Tri16 encodes an acyltransferase that catalyzes the formation of ester side groups at C 8 during trichothecene biosynthesis. We also report the presence of a Tri16 ortholog in Gibberella pulicaris R-6380 that is likely linked to a presumably inactive ortholog for Tri1. PMID- 14532048 TI - Utilization of acyl-homoserine lactone quorum signals for growth by a soil pseudomonad and Pseudomonas aeruginosa PAO1. AB - Acyl-homoserine lactones (AHLs) are employed by several Proteobacteria as quorum sensing signals. Past studies have established that these compounds are subject to biochemical decay and can be used as growth nutrients. Here we describe the isolation of a soil bacterium, Pseudomonas strain PAI-A, that degrades 3 oxododecanoyl-homoserine lactone (3OC12HSL) and other long-acyl, but not short acyl, AHLs as sole energy sources for growth. The small-subunit rRNA gene from strain PAI-A was 98.4% identical to that of Pseudomonas aeruginosa, but the soil isolate did not produce obvious pigments or AHLs or grow under denitrifying conditions or at 42 degrees C. The quorum-sensing bacterium P. aeruginosa, which produces both 3OC12HSL and C4HSL, was examined for the ability to utilize AHLs for growth. It did so with a specificity similar to that of strain PAI-A, i.e., degrading long-acyl but not short-acyl AHLs. In contrast to the growth observed with strain PAI-A, P. aeruginosa strain PAO1 growth on AHLs commenced only after extremely long lag phases. Liquid-chromatography-atmospheric pressure chemical ionization-mass spectrometry analyses indicate that strain PAO1 degrades long acyl AHLs via an AHL acylase and a homoserine-generating HSL lactonase. A P. aeruginosa gene, pvdQ (PA2385), has previously been identified as being a homologue of the AHL acylase described as occurring in a Ralstonia species. Escherichia coli expressing pvdQ catalyzed the rapid inactivation of long-acyl AHLs and the release of HSL. P. aeruginosa engineered to constitutively express pvdQ did not accumulate its 3OC12HSL quorum signal when grown in rich media. However, pvdQ knockout mutants of P. aeruginosa were still able to grow by utilizing 3OC12HSL. To our knowledge, this is the first report of the degradation of AHLs by pseudomonads or other gamma-Proteobacteria, of AHL acylase activity in a quorum-sensing bacterium, of HSL lactonase activity in any bacterium, and of AHL degradation with specificity only towards AHLs with long side chains. PMID- 14532049 TI - Comparing denitrification estimates for a Texas estuary by using acetylene inhibition and membrane inlet mass spectrometry. AB - Characterizing denitrification rates in aquatic ecosystems is essential to understanding how systems may respond to increased nutrient loading. Thus, it is important to ensure the precision and accuracy of the methods employed for measuring denitrification rates. The acetylene (C2H2) inhibition method is a simple technique for estimating denitrification. However, potential problems, such as inhibition of nitrification and incomplete inhibition of nitrous oxide reduction, may influence rate estimates. Recently, membrane inlet mass spectrometry (MIMS) has been used to measure denitrification in aquatic systems. Comparable results were obtained with MIMS and C2H2 inhibition methods when chloramphenicol was added to C2H2 inhibition assay mixtures to inhibit new synthesis of denitrifying enzymes. Dissolved-oxygen profiles indicated that surface layers of sediment cores subjected to the MIMS flowthrough incubation remained oxic whereas cores incubated using the C2H2 inhibition methods did not. Analysis of the microbial assemblages before and after incubations indicated significant changes in the sediment surface populations during the long flowthrough incubation for MIMS analysis but not during the shorter incubation used for the C2H2 inhibition method. However, bacterial community changes were also small in MIMS cores at the oxygen transition zone where denitrification occurs. The C2H2 inhibition method with chloramphenicol addition, conducted over short incubation intervals, provides a cost-effective method for estimating denitrification, and rate estimates are comparable to those obtained by the MIMS method. PMID- 14532050 TI - Cloning, characterization, and functional expression of the Klebsiella oxytoca xylodextrin utilization operon (xynTB) in Escherichia coli. AB - Escherichia coli is being developed as a biocatalyst for bulk chemical production from inexpensive carbohydrates derived from lignocellulose. Potential substrates include the soluble xylodextrins (xyloside, xylooligosaccharide) and xylobiose that are produced by treatments designed to expose cellulose for subsequent enzymatic hydrolysis. Adjacent genes encoding xylobiose uptake and hydrolysis were cloned from Klebsiella oxytoca M5A1 and are functionally expressed in ethanologenic E. coli. The xylosidase encoded by xynB contains the COG3507 domain characteristic of glycosyl hydrolase family 43. The xynT gene encodes a membrane protein containing the MelB domain (COG2211) found in Na(+)/melibiose symporters and related proteins. These two genes form a bicistronic operon that appears to be regulated by xylose (XylR) and by catabolite repression in both K. oxytoca and recombinant E. coli. Homologs of this operon were found in Klebsiella pneumoniae, Lactobacillus lactis, E. coli, Clostridium acetobutylicum, and Bacillus subtilis based on sequence comparisons. Based on similarities in protein sequence, the xynTB genes in K. oxytoca appear to have originated from a gram-positive ancestor related to L. lactis. Functional expression of xynB allowed ethanologenic E. coli to metabolize xylodextrins (xylosides) containing up to six xylose residues without the addition of enzyme supplements. 4-O-methylglucuronic acid substitutions at the nonreducing termini of soluble xylodextrins blocked further degradation by the XynB xylosidase. The rate of xylodextrin utilization by recombinant E. coli was increased when a full-length xynT gene was included with xynB, consistent with xynT functioning as a symport. Hydrolysis rates were inversely related to xylodextrin chain length, with xylobiose as the preferred substrate. Xylodextrins were utilized more rapidly by recombinant E. coli than K. oxytoca M5A1 (the source of xynT and xynB). XynB exhibited weak arabinosidase activity, 3% that of xylosidase. PMID- 14532051 TI - Bacterial chemotaxis to naphthalene desorbing from a nonaqueous liquid. AB - Bacterial chemotaxis has the potential to increase the rate of degradation of chemoattractants, but its influence on degradation of hydrophobic attractants initially dissolved in a non-aqueous-phase liquid (NAPL) has not been examined. We studied the effect of chemotaxis by Pseudomonas putida G7 on naphthalene mass transfer and degradation in a system in which the naphthalene was dissolved in a model NAPL. Chemotaxis by wild-type P. putida G7 increased the rates of naphthalene desorption and degradation relative to rates observed with nonchemotactic and nonmotile mutant strains. While biodegradation alone influenced the rate of substrate desorption by increasing the concentration gradient against which desorption occurred, chemotaxis created an even steeper gradient as the cells accumulated near the NAPL source. The extent to which chemotaxis affected naphthalene desorption and degradation depended on the initial bacterial and naphthalene concentrations, reflecting the influences of these variables on concentration gradients and on the relative rates of mass transfer and biodegradation. The results of this study suggest that chemotaxis can substantially increase the rates of mass transfer and degradation of NAPL associated hydrophobic pollutants. PMID- 14532052 TI - Community composition and functioning of denitrifying bacteria from adjacent meadow and forest soils. AB - We investigated communities of denitrifying bacteria from adjacent meadow and forest soils. Our objectives were to explore spatial gradients in denitrifier communities from meadow to forest, examine whether community composition was related to ecological properties (such as vegetation type and process rates), and determine phylogenetic relationships among denitrifiers. nosZ, a key gene in the denitrification pathway for nitrous oxide reductase, served as a marker for denitrifying bacteria. Denitrifying enzyme activity (DEA) was measured as a proxy for function. Other variables, such as nitrification potential and soil C/N ratio, were also measured. Soil samples were taken along transects that spanned meadow-forest boundaries at two sites in the H. J. Andrews Experimental Forest in the Western Cascade Mountains of Oregon. Results indicated strong functional and structural community differences between the meadow and forest soils. Levels of DEA were an order of magnitude higher in the meadow soils. Denitrifying community composition was related to process rates and vegetation type as determined on the basis of multivariate analyses of nosZ terminal restriction fragment length polymorphism profiles. Denitrifier communities formed distinct groups according to vegetation type and site. Screening 225 nosZ clones yielded 47 unique denitrifying genotypes; the most dominant genotype occurred 31 times, and half the genotypes occurred once. Several dominant and less-dominant denitrifying genotypes were more characteristic of either meadow or forest soils. The majority of nosZ fragments sequenced from meadow or forest soils were most similar to nosZ from the Rhizobiaceae group in alpha-Proteobacteria species. Denitrifying community composition, as well as environmental factors, may contribute to the variability of denitrification rates in these systems. PMID- 14532053 TI - Identification and characterization of the CYP52 family of Candida tropicalis ATCC 20336, important for the conversion of fatty acids and alkanes to alpha,omega-dicarboxylic acids. AB - Candida tropicalis ATCC 20336 excretes alpha,omega-dicarboxylic acids as a by product when cultured on n-alkanes or fatty acids as the carbon source. Previously, a beta-oxidation-blocked derivative of ATCC 20336 was constructed which showed a dramatic increase in the production of dicarboxylic acids. This paper describes the next steps in strain improvement, which were directed toward the isolation and characterization of genes encoding the omega-hydroxylase enzymes catalyzing the first step in the omega-oxidation pathway. Cytochrome P450 monooxygenase (CYP) and the accompanying NADPH cytochrome P450 reductase (NCP) constitute the hydroxylase complex responsible for the first and rate-limiting step of omega-oxidation of n-alkanes and fatty acids. 10 members of the alkane inducible P450 gene family (CYP52) of C. tropicalis ATCC20336 as well as the accompanying NCP were cloned and sequenced. The 10 CYP genes represent four unique genes with their putative alleles and two unique genes for which no allelic variant was identified. Of the 10 genes, CYP52A13 and CYP52A14 showed the highest levels of mRNA induction, as determined by quantitative competitive reverse transcription-PCR during fermentation with pure oleic fatty acid (27-fold increase), pure octadecane (32-fold increase), and a mixed fatty acid feed, Emersol 267 (54-fold increase). The allelic pair CYP52A17 and CYP52A18 was also induced under all three conditions but to a lesser extent. Moderate induction of CYP52A12 was observed. These results identify the CYP52 and NCP genes as being involved in alpha,omega-dicarboxylic acid production by C. tropicalis and provide the foundation for biocatalyst improvement. PMID- 14532055 TI - Compound-specific isotopic fractionation patterns suggest different carbon metabolisms among Chloroflexus-like bacteria in hot-spring microbial mats. AB - Stable carbon isotope fractionations between dissolved inorganic carbon and lipid biomarkers suggest photoautotrophy by Chloroflexus-like organisms in sulfidic and nonsulfidic Yellowstone hot springs. Where co-occurring, cyanobacteria appear to cross-feed Chloroflexus-like organisms supporting photoheterotrophy as well, although the relatively small 13C fractionation associated with cyanobacterial sugar biosynthesis may sometimes obscure this process. PMID- 14532054 TI - Transformation of fatty acids catalyzed by cytochrome P450 monooxygenase enzymes of Candida tropicalis. AB - Candida tropicalis ATCC 20336 can grow on fatty acids or alkanes as its sole source of carbon and energy, but strains blocked in beta-oxidation convert these substrates to long-chain alpha,omega-dicarboxylic acids (diacids), compounds of potential commercial value (Picataggio et al., Biotechnology 10:894-898, 1992). The initial step in the formation of these diacids, which is thought to be rate limiting, is omega-hydroxylation by a cytochrome P450 (CYP) monooxygenase. C. tropicalis ATCC 20336 contains a family of CYP genes, and when ATCC 20336 or its derivatives are exposed to oleic acid (C(18:1)), two cytochrome P450s, CYP52A13 and CYP52A17, are consistently strongly induced (Craft et al., this issue). To determine the relative activity of each of these enzymes and their contribution to diacid formation, both cytochrome P450s were expressed separately in insect cells in conjunction with the C. tropicalis cytochrome P450 reductase (NCP). Microsomes prepared from these cells were analyzed for their ability to oxidize fatty acids. CYP52A13 preferentially oxidized oleic acid and other unsaturated acids to omega-hydroxy acids. CYP52A17 also oxidized oleic acid efficiently but converted shorter, saturated fatty acids such as myristic acid (C(14:0)) much more effectively. Both enzymes, in particular CYP52A17, also oxidized omega hydroxy fatty acids, ultimately generating the alpha,omega-diacid. Consideration of these different specificities and selectivities will help determine which enzymes to amplify in strains blocked for beta-oxidation to enhance the production of dicarboxylic acids. The activity spectrum also identified other potential oxidation targets for commercial development. PMID- 14532056 TI - Phylogenetic diversity, abundance, and axial distribution of bacteria in the intestinal tract of two soil-feeding termites (Cubitermes spp.). AB - The hindgut of soil-feeding termites is highly compartmentalized and characterized by pronounced axial dynamics of the intestinal pH and microbial processes such as hydrogen production, methanogenesis, and reductive acetogenesis. Nothing is known about the bacterial diversity and the abundance or axial distribution of the major phylogenetic groups in the different gut compartments. In this study, we showed that the variety of physicochemical conditions is reflected in the diversity of the microbial communities in the different gut compartments of two Cubitermes species (TERMITIDAE: Termitinae). 16S rRNA gene clones from the highly alkaline first proctodeal segment (P1) of Cubitermes orthognathus represented almost exclusively gram-positive bacteria with low G+C content (LGC bacteria). In the posterior gut segments, their proportion decreased progressively, and the clone libraries comprised a variety of phyla, including the Cytophaga-Flexibacter-Bacteroides group, various subgroups of Proteobacteria, and the spirochetes. Phylogenetic analysis revealed that many of the clones clustered with sequences from the guts of other termites, and some even formed clusters containing only clones from C. orthognathus. The abundance and axial distribution of major phylogenetic groups in the gut of Cubitermes ugandensis were determined by fluorescence in situ hybridization with group-specific oligonucleotide probes. While the results were generally in good agreement with those of the clonal analysis, direct counts with probes specific for the Planctomycetales revealed a severe underestimation of representatives of this phylum in the clone libraries. Results obtained with newly designed FISH probes directed against two clusters of LGC clones from C. orthognathus indicated that the clones were restricted to specific gut regions. A molecular fingerprinting analysis published in a companion paper (D. Schmitt-Wagner, M. W. Friedrich, B. Wagner, and A. Brune, Appl. Environ. Microbiol. 69:6018-6024, 2003) corroborated the presence of compartment-specific bacterial communities in the gut of different Cubitermes species. PMID- 14532057 TI - Axial dynamics, stability, and interspecies similarity of bacterial community structure in the highly compartmentalized gut of soil-feeding termites (Cubitermes spp.). AB - The highly compartmentalized gut of soil-feeding termites is characterized by pronounced axial dynamics in physicochemical conditions and microbial processes. In a companion paper (D. Schmitt-Wagner, M. W. Friedrich, B. Wagner, and A. Brune, Appl. Environ. Microbiol. 69:6007-6017, 2003), we demonstrated that the variety of physicochemical conditions in the different gut compartments of Cubitermes spp. is reflected in the diversity of the respective intestinal microbial communities. Here, we used molecular fingerprints of 16S rRNA genes of the bacterial community, obtained by terminal restriction fragment length polymorphism (T-RFLP) analysis, to describe the axial dynamics of the bacterial community structure in the different gut sections. Comparison of the T-RFLP profiles with the predicted terminal restriction fragments of the clones in clone libraries of the gut segments in Cubitermes orthognathus confirmed that all hindgut sections harbored distinct bacterial communities. Morisita indices of community similarity, calculated by comparing the different patterns, revealed large differences between the bacterial communities of soil, gut, and nest material and also among the individual gut sections. By contrast, comparison of the homologous gut segments of different Cubitermes species indicated that the three termite species investigated possessed a similar, gut-specific microbiota that remained comparatively stable even during several months of maintenance in the laboratory. PMID- 14532058 TI - Effect of pH and oxalate on hydroquinone-derived hydroxyl radical formation during brown rot wood degradation. AB - The redox cycle of 2,5-dimethoxybenzoquinone (2,5-DMBQ) is proposed as a source of reducing equivalent for the regeneration of Fe2+ and H2O2 in brown rot fungal decay of wood. Oxalate has also been proposed to be the physiological iron reductant. We characterized the effect of pH and oxalate on the 2,5-DMBQ-driven Fenton chemistry and on Fe3+ reduction and oxidation. Hydroxyl radical formation was assessed by lipid peroxidation. We found that hydroquinone (2,5-DMHQ) is very stable in the absence of iron at pH 2 to 4, the pH of degraded wood. 2,5-DMHQ readily reduces Fe3+ at a rate constant of 4.5 x 10(3) M(-1)s(-1) at pH 4.0. Fe2+ is also very stable at a low pH. H2O2 generation results from the autoxidation of the semiquinone radical and was observed only when 2,5-DMHQ was incubated with Fe3+. Consistent with this conclusion, lipid peroxidation occurred only in incubation mixtures containing both 2,5-DMHQ and Fe3+. Catalase and hydroxyl radical scavengers were effective inhibitors of lipid peroxidation, whereas superoxide dismutase caused no inhibition. At a low concentration of oxalate (50 micro M), ferric ion reduction and lipid peroxidation are enhanced. Thus, the enhancement of both ferric ion reduction and lipid peroxidation may be due to oxalate increasing the solubility of the ferric ion. Increasing the oxalate concentration such that the oxalate/ferric ion ratio favored formation of the 2:1 and 3:1 complexes resulted in inhibition of iron reduction and lipid peroxidation. Our results confirm that hydroxyl radical formation occurs via the 2,5-DMBQ redox cycle. PMID- 14532059 TI - ISLpl1 is a functional IS30-related insertion element in Lactobacillus plantarum that is also found in other lactic acid bacteria. AB - We describe the first functional insertion sequence (IS) element in Lactobacillus plantarum. ISLpl1, an IS30-related element, was found on the pLp3 plasmid in strain FB335. By selection of spontaneous mutants able to grow in the presence of uracil, it was demonstrated that the IS had transposed into the uracil phosphoribosyltransferase-encoding gene upp on the FB335 chromosome. The plasmid carried IS element was also sequenced, and a second potential IS element was found: ISLpl2, an IS150-related element adjacent to ISLpl1. When Southern hybridization was used, the copy number and genome (plasmid versus chromosome) distribution data revealed different numbers and patterns of ISLpl1-related sequences in different L. plantarum strains as well as in Pediococcus strains. The ISLpl1 pattern changed over many generations of the strain L. plantarum NCIMB 1406. This finding strongly supports our hypothesis that ISLpl1 is a mobile element in L. plantarum. Database analysis revealed five quasi-identical ISLpl1 elements in Lactobacillus, Pediococcus, and Oenococcus strains. Three of these elements may be cryptic IS, since point mutations or 1-nucleotide deletions were found in their transposase-encoding genes. In some cases, ISLpl1 was linked to genes involved in cold shock adaptation, bacteriocin production, sugar utilization, or antibiotic resistance. ISLpl1 is transferred among lactic acid bacteria (LAB) and may play a role in LAB genome plasticity and adaptation to their environment. PMID- 14532060 TI - Distribution of the coenzyme M pathway of epoxide metabolism among ethene- and vinyl chloride-degrading Mycobacterium strains. AB - An epoxyalkane:coenzyme M (CoM) transferase (EaCoMT) enzyme was recently found to be active in the aerobic vinyl chloride (VC) and ethene assimilation pathways of Mycobacterium strain JS60. In the present study, EaCoMT activity and genes were investigated in 10 different mycobacteria isolated on VC or ethene from diverse environmental samples. In all cases, epoxyethane metabolism in cell extracts was dependent on CoM, with average specific activities of EaCoMT between 380 and 2,910 nmol/min/mg of protein. PCR with primers based on conserved regions of EaCoMT genes from Mycobacterium strain JS60 and the propene oxidizers Xanthobacter strain Py2 and Rhodococcus strain B-276 yielded fragments (834 bp) of EaCoMT genes from all of the VC- and ethene-assimilating isolates. The Mycobacterium EaCoMT genes form a distinct cluster and are more closely related to the EaCoMT of Rhodococcus strain B-276 than that of Xanthobacter strain Py2. The incongruence of the EaCoMT and 16S rRNA gene trees and the fact that isolates from geographically distant locations possessed almost identical EaCoMT genes suggest that lateral transfer of EaCoMT among the Mycobacterium strains has occurred. Pulsed-field gel electrophoresis revealed large linear plasmids (110 to 330 kb) in all of the VC-degrading strains. In Southern blotting experiments, the strain JS60 EaCoMT gene hybridized to many of the plasmids. The CoM-mediated pathway of epoxide metabolism appears to be universal in alkene-assimilating mycobacteria, possibly because of plasmid-mediated lateral gene transfer. PMID- 14532062 TI - Coaggregation among nonflocculating bacteria isolated from activated sludge. AB - Thirty-two strains of nonflocculating bacteria isolated from sewage-activated sludge were tested by a spectrophotometric assay for their ability to coaggregate with one other in two-membered systems. Among these strains, eight showed significant (74 to 99%) coaggregation with Acinetobacter johnsonii S35 while only four strains coaggregated, to a lesser extent (43 to 65%), with Acinetobacter junii S33. The extent and pattern of coaggregation as well as the aggregate size showed good correlation with cellular characteristics of the coaggregating partners. These strains were identified by sequencing of full-length 16S rRNA genes. A. johnsonii S35 could coaggregate with strains of several genera, such as Oligotropha carboxidovorans, Microbacterium esteraromaticum, and Xanthomonas spp. The role of Acinetobacter isolates as bridging organisms in multigeneric coaggregates is indicated. This investigation revealed the role of much-neglected nonflocculating bacteria in floc formation in activated sludge. PMID- 14532061 TI - Lysine-2,3-aminomutase and beta-lysine acetyltransferase genes of methanogenic archaea are salt induced and are essential for the biosynthesis of Nepsilon acetyl-beta-lysine and growth at high salinity. AB - The compatible solute N(epsilon)-acetyl-beta-lysine is unique to methanogenic archaea and is produced under salt stress only. However, the molecular basis for the salt-dependent regulation of N(epsilon)-acetyl-beta-lysine formation is unknown. Genes potentially encoding lysine-2,3-aminomutase (ablA) and beta-lysine acetyltransferase (ablB), which are assumed to catalyze N(epsilon)-acetyl-beta lysine formation from alpha-lysine, were identified on the chromosomes of the methanogenic archaea Methanosarcina mazei Go1, Methanosarcina acetivorans, Methanosarcina barkeri, Methanococcus jannaschii, and Methanococcus maripaludis. The order of the two genes was identical in the five organisms, and the deduced proteins were very similar, indicating a high degree of conservation of structure and function. Northern blot analysis revealed that the two genes are organized in an operon (termed the abl operon) in M. mazei Go1. Expression of the abl operon was strictly salt dependent. The abl operon was deleted in the genetically tractable M. maripaludis. Delta(abl) mutants of M. maripaludis no longer produced N(epsilon)-acetyl-beta-lysine and were incapable of growth at high salt concentrations, indicating that the abl operon is essential for N(epsilon)-acetyl beta-lysine synthesis. These experiments revealed the first genes involved in the biosynthesis of compatible solutes in methanogens. PMID- 14532063 TI - Optimized expression of a thermostable xylanase from Thermomyces lanuginosus in Pichia pastoris. AB - Highly efficient production of a Thermomyces lanuginosus IOC-4145 beta-1,4 xylanase was achieved in Pichia pastoris under the control of the AOX1 promoter. P. pastoris colonies expressing recombinant xylanase were selected by enzymatic activity plate assay, and their ability to secrete high levels of the enzyme was evaluated in small-scale cultures. Furthermore, an optimization of enzyme production was carried out with a 2(3) factorial design. The influence of initial cell density, methanol, and yeast nitrogen base concentration was evaluated, and initial cell density was found to be the most important parameter. A time course profile of recombinant xylanase production in 1-liter flasks with the optimized conditions was performed and 148 mg of xylanase per liter was achieved. Native and recombinant xylanases were purified by gel filtration and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, circular dichroism spectroscopy, matrix-assisted laser desorption ionization-time of flight-mass spectrometry and physicochemical behavior. Three recombinant protein species of 21.9, 22.1, and 22.3 kDa were detected in the mass spectrum due to variability in the amino terminus. The optimum temperature, thermostability, and circular dichroic spectra of the recombinant and native xylanases were identical. For both enzymes, the optimum temperature was 75 degrees C, and they retained 60% of their original activity after 80 min at 70 degrees C or 40 min at 80 degrees C. The high level of fully active recombinant xylanase obtained in P. pastoris makes this expression system attractive for fermentor growth and industrial applications. PMID- 14532064 TI - Molecular diversity of sulfate-reducing bacteria from two different continental margin habitats. AB - This study examined the natural diversity and distributions of sulfate-reducing bacteria along a natural carbon gradient extending down the shelf-slope transition zone of the eastern Pacific continental margin. Dissimilatory (bi)sulfite reductase gene sequences (dsrAB) were PCR amplified and cloned from five different sampling sites, each at a discrete depth, from two different margin systems, one off the Pacific coast of Mexico and another off the coast of Washington State. A total of 1,762 clones were recovered and evaluated by restriction fragment length polymorphism (RFLP) analysis. The majority of the gene sequences recovered showed site and depth restricted distributions; however, a limited number of gene sequences were widely distributed within and between the margin systems. Cluster analysis identified 175 unique RFLP patterns, and nucleotide sequences were determined for corresponding clones. Several different continental margin DsrA sequences clustered with those from formally characterized taxa belonging to the delta subdivision of the class Proteobacteria (Desulfobulbus propionicus, Desulfosarcina variabilis) and the Bacillus Clostridium (Desulfotomaculum putei) divisions, although the majority of the recovered sequences were phylogenetically divergent relative to all of the other DsrA sequences available for comparison. This study revealed extensive new genetic diversity among sulfate-reducing bacteria in continental margin sedimentary habitats, which appears to be tightly coupled to slope depth, specifically carbon bioavailability. PMID- 14532065 TI - Diversity of Wolbachia endosymbionts in heteropteran bugs. AB - An extensive survey of Wolbachia endosymbionts in Japanese terrestrial heteropteran bugs was performed by PCR detection with universal primers for wsp and ftsZ genes of Wolbachia, cloning of the PCR products, restriction fragment length polymorphism analysis of infecting Wolbachia types, and molecular phylogenetic characterization of all the detected Wolbachia strains. Of 134 heteropteran species from 19 families examined, Wolbachia infection was detected in 47 species from 13 families. From the 47 species, 59 Wolbachia strains were identified. Of the 59 strains, 16 and 43 were assigned to A group and B group in the Wolbachia phylogeny, respectively. The 47 species of Wolbachia-infected bugs were classified into 8 species with A infection, 28 species with B infection, 2 species with AA infection, 3 species with AB infection, 5 species with BB infection, and 1 species with ABB infection. Molecular phylogenetic analysis showed little congruence between Wolbachia phylogeny and host systematics, suggesting frequent horizontal transfers of Wolbachia in the evolutionary course of the Heteroptera. The phylogenetic analysis also revealed several novel lineages of Wolbachia. Based on statistical analyses of the multiple infections, we propose a hypothetical view that, in the heteropteran bugs, interactions between coinfecting Wolbachia strains are generally not intense and that Wolbachia coinfections have been established through a stochastic process probably depending on occasional horizontal transfers. PMID- 14532066 TI - Influence of nutritional factors on the nature, yield, and composition of exopolysaccharides produced by Gluconacetobacter xylinus I-2281. AB - The influence of substrate composition on the yield, nature, and composition of exopolysaccharides (EPS) produced by the food-grade strain Gluconacetobacter xylinus I-2281 was investigated during controlled cultivations on mixed substrates containing acetate and either glucose, sucrose, or fructose. Enzymatic activity analysis and acid hydrolysis revealed that two EPS, gluconacetan and levan, were produced by G. xylinus. In contrast to other acetic acid strains, no exocellulose formation has been measured. Considerable differences in metabolite yields have been observed with regard to the carbohydrate source. It was shown that glucose was inadequate for EPS production since most of this substrate (0.84 C-mol/C-mol) was oxidized into gluconic acid, 2-ketogluconic acid, and 5 ketogluconic acid. In contrast, sucrose and fructose supported a 0.35 C-mol/C-mol gluconacetan yield. In addition, growing G. xylinus on sucrose produced a 0.07 C mol/C-mol levan yield. The composition of EPS remained unchanged during the course of the fermentations. Levan sucrase activity was found to be mainly membrane associated. In addition to levan production, an analysis of levan sucrase's activity also explained the formation of glucose oxides during fermentation on sucrose through the release of glucose. The biosynthetic pathway of gluconacetan synthesis has also been explored. Although the activity of key enzymes showed large differences to be a function of the carbon source, the ratio of their activities remained similar from one carbon source to another and corresponded to the ratio of precursor needs as deduced from the gluconacetan composition. PMID- 14532067 TI - Molecular analysis of the rfb O antigen gene cluster of Salmonella enterica serogroup O:6,14 and development of a serogroup-specific PCR assay. AB - The Kauffmann-White scheme for serotyping Salmonella recognizes 46 somatic (O) antigen groups, which together with detection of the flagellar (H) antigens form the basis for serotype identification. Although serotyping has become an invaluable typing method for epidemiological investigations of Salmonella, it does have some practical limitations. We have been characterizing the genes required for O and H antigen biosynthesis with the goal of developing a DNA-based system for the determination of serotype in Salmonella. The majority of the enzymes involved in O antigen biosynthesis are encoded by the rfb gene cluster. We report the sequencing of the rfb region from S. enterica serotype Sundsvall (serogroup O:6,14). The S. enterica serotype Sundsvall rfb region is 8.4 kb in length and comprises six open reading frames. When compared with other previously characterized rfb regions, the serogroup O:6,14 sequence is most related to serogroup C(1). On the basis of DNA sequence similarity, we identified two genes from the mannose biosynthetic pathway, two mannosyl transferase genes, the O unit flippase gene and, possibly, the O antigen polymerase. The whole cluster is derived from a low-G+C-content organism. Comparative sequencing of an additional serogroup O:6,14 isolate (S. enterica serotype Carrau) revealed a highly homologous sequence, suggesting that O antigen factors O:24 and O:25 (additional O factors associated with serogroup O:6,14) are encoded outside the rfb gene cluster. We developed a serogroup O:6,14-specific PCR assay based on a region of the putative wzx (O antigen flippase) gene. This provides the basis for a sensitive and specific test for the rapid identification of Salmonella serogroup O:6,14. PMID- 14532068 TI - Escherichia coli ghost production by expression of lysis gene E and Staphylococcal nuclease. AB - The production of bacterial ghosts from Escherichia coli is accomplished by the controlled expression of phage phiX174 lysis gene E and, in contrast to other gram-negative bacterial species, is accompanied by the rare detection of nonlysed, reproductive cells within the ghost preparation. To overcome this problem, the expression of a secondary killing gene was suggested to give rise to the complete genetic inactivation of the bacterial samples. The expression of staphylococcal nuclease A in E. coli resulted in intracellular accumulation of the protein and degradation of the host DNA into fragments shorter than 100 bp. Two expression systems for the nuclease are presented and were combined with the protein E-mediated lysis system. Under optimized conditions for the coexpression of gene E and the staphylococcal nuclease, the concentration of viable cells fell below the lower limit of detection, whereas the rates of ghost formation were not affected. With regard to the absence of reproductive cells from the ghost fractions, the reduction of viability could be determined as being at least 7 to 8 orders of magnitude. The lysis process was characterized by electrophoretic analysis and absolute quantification of the genetic material within the cells and the culture supernatant via real-time PCR. The ongoing degradation of the bacterial nucleic acids resulted in a continuous quantitative clearance of the genetic material associated with the lysing cells until the concentrations fell below the detection limits of either assay. No functional, released genetic units (genes) were detected within the supernatant during the lysis process, including nuclease expression. PMID- 14532069 TI - RpoS-dependent stress response and exoenzyme production in Vibrio vulnificus. AB - Vibrio vulnificus is an estuarine bacterium capable of causing rapidly fatal infections through both ingestion and wound infection. Like other opportunistic pathogens, V. vulnificus must adapt to potentially stressful environmental changes while living freely in seawater, upon colonization of the oyster gut, and upon infection of such diverse hosts as humans and eels. In order to begin to understand the ability of V. vulnificus to respond to such stresses, we examined the role of the alternate sigma factor RpoS, which is important in stress response and virulence in many pathogens. An rpoS mutant of V. vulnificus strain C7184o was constructed by homologous recombination. The mutant strain exhibited a decreased ability to survive diverse environmental stresses, including exposure to hydrogen peroxide, hyperosmolarity, and acidic conditions. The most striking difference was a high sensitivity of the mutant to hydrogen peroxide. Albuminase, caseinase, and elastase activity were detected in the wild type but not in the mutant strain, and an additional two hydrolytic activities (collagenase and gelatinase) were reduced in the mutant strain compared to the wild type. Additionally, the motility of the rpoS mutant was severely diminished. Overall, these studies suggest that rpoS in V. vulnificus is important for adaptation to environmental changes and may have a role in virulence. PMID- 14532070 TI - Marker rescue studies of the transfer of recombinant DNA to Streptococcus gordonii in vitro, in foods and gnotobiotic rats. AB - A plasmid marker rescue system based on restoration of the nptII gene was established in Streptococcus gordonii to study the transfer of bacterial and transgenic plant DNA by transformation. In vitro studies revealed that the marker rescue efficiency depends on the type of donor DNA. Plasmid and chromosomal DNA of bacteria as well as DNA of transgenic potatoes were transferred with efficiencies ranging from 8.1 x 10(-6) to 5.8 x 10(-7) transformants per nptII gene. Using a 792-bp amplification product of nptII the efficiency was strongly decreased (9.8 x 10(-9)). In blood sausage, marker rescue using plasmid DNA was detectable (7.9 x 10(-10)), whereas in milk heat-inactivated horse serum (HHS) had to be added to obtain an efficiency of 2.7 x 10(-11). No marker rescue was detected in extracts of transgenic potatoes despite addition of HHS. In vivo transformation of S. gordonii LTH 5597 was studied in monoassociated rats by using plasmid DNA. No marker rescue could be detected in vivo, although transformation was detected in the presence of saliva and fecal samples supplemented with HHS. It was also shown that plasmid DNA persists in rat saliva permitting transformation for up to 6 h of incubation. It is suggested that the lack of marker rescue is due to the absence of competence-stimulating factors such as serum proteins in rat saliva. PMID- 14532071 TI - Formaldehyde fixation contributes to detoxification for growth of a nonmethylotroph, Burkholderia cepacia TM1, on vanillic acid. AB - During bacterial degradation of methoxylated lignin monomers, such as vanillin and vanillic acid, formaldehyde is released through the reaction catalyzed by vanillic acid demethylase. When Burkholderia cepacia TM1 was grown on vanillin or vanillic acid as the sole carbon source, the enzymes 3-hexulose-6-phosphate synthase (HPS) and 6-phospho-3-hexuloisomerase (PHI) were induced. These enzymes were also expressed during growth on Luria-Bertani medium containing formaldehyde. To understand the roles of these enzymes, the hps and phi genes from a methylotrophic bacterium, Methylomonas aminofaciens 77a, were introduced into B. cepacia TM1. The transformant strain constitutively expressed the genes for HPS and PHI, and these activities were two- or threefold higher than the activities in the wild strain. Incorporation of [14C]formaldehyde into the cell constituents was increased by overexpression of the genes. Furthermore, the degradation of vanillic acid and the growth yield were significantly improved at a high concentration of vanillic acid (60 mM) in the transformant strain. These results suggest that HPS and PHI play significant roles in the detoxification and assimilation of formaldehyde. This is the first report that enhancement of the HPS/PHI pathway could improve the degradation of vanillic acid in nonmethylotrophic bacteria. PMID- 14532072 TI - Characterization of multiple-substrate utilization by anthracene-degrading Mycobacterium frederiksbergense LB501T. AB - Stable carbon isotope analysis of biomass and analyses of phospholipid fatty acids (PLFA), glycolipid fatty acids (GLFA), and mycolic acids were used to characterize mixed-substrate utilization by Mycobacterium frederiksbergense LB501T under various substrate regimens. The distinct (13)C contents of anthracene and glucose as representatives of typical hydrophobic pollutants and naturally occurring organic compounds, respectively, were monitored during formation into biomass and used to quantify the relative contributions of the two carbon sources to biomass formation. Moreover, the influence of mixed-substrate utilization on PLFA, GLFA, and mycolic acid profiles and cell surface hydrophobicity was investigated. Results revealed that M. frederiksbergense LB501T degrades anthracene and forms biomass from it even in the presence of more readily available dissolved glucose. The relative ratios of straight-chain saturated PLFA to the corresponding unsaturated PLFA and the total fraction of saturated cyclopropyl-branched PLFA of M. frederiksbergense LB501T depended on the carbon source and the various rates of addition of mixed substrates, whereas no such trend was observed with GLFA. Higher proportions of anthracene in the carbon source mixture led to higher cell surface hydrophobicities and more hydrophobic mycolic acids, which in turn appeared to be valuable indicators for substrate utilization by M. frederiksbergense LB501T. The capability of polycyclic aromatic hydrocarbon (PAH)-degrading bacteria to utilize readily available substrates besides the poorly available PAHs favors the buildup of PAH degrading biomass. Feeding of supplementary carbon substrates may therefore promote bioremediation, provided that it sustains the pollutant-degrading population rather than other members of the microbial community. PMID- 14532073 TI - Terminal restriction fragment length polymorphism data analysis. PMID- 14532074 TI - Radioisotopic, culture-based, and oligonucleotide microchip analyses of thermophilic microbial communities in a continental high-temperature petroleum reservoir. AB - Activity measurements by radioisotopic methods and cultural and molecular approaches were used in parallel to investigate the microbial biodiversity and its physiological potential in formation waters of the Samotlor high-temperature oil reservoir (Western Siberia, Russia). Sulfate reduction with rates not exceeding 20 nmol of H(2)S liter(-1) day(-1) occurred at 60 and 80 degrees C. In upper horizons (AB, A, and B), methanogenesis (lithotrophic and/or acetoclastic) was detected only in wells in which sulfate reduction did not occur. In some of the wells from deeper (J) horizons, high-temperature sulfate reduction and methanogenesis occurred simultaneously, the rate of lithotrophic methanogenesis exceeding 80 nmol of CH(4) liter(-1) day(-1). Enrichment cultures indicated the presence of diverse physiological groups representing aerobic and anaerobic thermophiles and hyperthermophiles; fermentative organotrophs were predominant. Phylogenetic analyses of 15 isolates identified representatives of the genera Thermotoga, Thermoanaerobacter, Geobacillus, Petrotoga, Thermosipho, and Thermococcus, the latter four being represented by new species. Except for Thermosipho, the isolates were members of genera recovered earlier from similar habitats. DNA obtained from three samples was hybridized with a set of oligonucleotide probes targeting selected microbial groups encompassing key genera of thermophilic bacteria and archaea. Oligonucleotide microchip analyses confirmed the cultural data but also revealed the presence of several groups of microorganisms that escaped cultivation, among them representatives of the Aquificales/Desulfurobacterium-Thermovibrio cluster and of the genera Desulfurococcus and Thermus, up to now unknown in this habitat. The unexpected presence of these organisms suggests that their distribution may be much wider than suspected. PMID- 14532075 TI - Patterns of community change among ammonia oxidizers in meadow soils upon long term incubation at different temperatures. AB - The effect of temperature on the community structure of ammonia-oxidizing bacteria was investigated in three different meadow soils. Two of the soils (OMS and GMS) were acidic (pH 5.0 to 5.8) and from sites in Germany with low annual mean temperature (about 10 degrees C), while KMS soil was slightly alkaline (pH 7.9) and from a site in Israel with a high annual mean temperature (about 22 degrees C). The soils were fertilized and incubated for up to 20 weeks in a moist state and as a buffered (pH 7) slurry amended with urea at different incubation temperatures (4 to 37 degrees C). OMS soil was also incubated with less fertilizer than the other soils. The community structure of ammonia oxidizers was analyzed before and after incubation by denaturing gradient gel electrophoresis (DGGE) of the amoA gene, which codes for the alpha subunit of ammonia monooxygenase. All amoA gene sequences found belonged to the genus Nitrosospira. The analysis showed community change due to temperature both in moist soil and in the soil slurry. Two patterns of community change were observed. One pattern was a change between the different Nitrosospira clusters, which was observed in moist soil and slurry incubations of GMS and OMS. Nitrosospira AmoA cluster 1 was mainly detected below 30 degrees C, while Nitrosospira cluster 4 was predominant at 25 degrees C. Nitrosospira clusters 3a, 3b, and 9 dominated at 30 degrees C. The second pattern, observed in KMS, showed a community shift predominantly within a single Nitrosospira cluster. The sequences of the individual DGGE bands that exhibited different trends with temperature belonged almost exclusively to Nitrosospira cluster 3a. We conclude that ammonia oxidizer populations are influenced by temperature. In addition, we confirmed previous observations that N fertilizer also influences the community structure of ammonia oxidizers. Thus, Nitrosospira cluster 1 was absent in OMS soil treated with less fertilizer, while Nitrosospira cluster 9 was only found in the sample given less fertilizer. PMID- 14532077 TI - Immobilization of arsenite and ferric iron by Acidithiobacillus ferrooxidans and its relevance to acid mine drainage. AB - Weathering of the As-rich pyrite-rich tailings of the abandoned mining site of Carnoules (southeastern France) results in the formation of acid waters heavily loaded with arsenic. Dissolved arsenic present in the seepage waters precipitates within a few meters from the bottom of the tailing dam in the presence of microorganisms. An Acidithiobacillus ferrooxidans strain, referred to as CC1, was isolated from the effluents. This strain was able to remove arsenic from a defined synthetic medium only when grown on ferrous iron. This A. ferrooxidans strain did not oxidize arsenite to arsenate directly or indirectly. Strain CC1 precipitated arsenic unexpectedly as arsenite but not arsenate, with ferric iron produced by its energy metabolism. Furthermore, arsenite was almost not found adsorbed on jarosite but associated with a poorly ordered schwertmannite. Arsenate is known to efficiently precipitate with ferric iron and sulfate in the form of more or less ordered schwertmannite, depending on the sulfur-to-arsenic ratio. Our data demonstrate that the coprecipitation of arsenite with schwertmannite also appears as a potential mechanism of arsenite removal in heavily contaminated acid waters. The removal of arsenite by coprecipitation with ferric iron appears to be a common property of the A. ferrooxidans species, as such a feature was observed with one private and three collection strains, one of which was the type strain. PMID- 14532078 TI - A model to estimate the optimal sample size for microbiological surveys. AB - Estimating optimal sample size for microbiological surveys is a challenge for laboratory managers. When insufficient sampling is conducted, biased inferences are likely; however, when excessive sampling is conducted valuable laboratory resources are wasted. This report presents a statistical model for the estimation of the sample size appropriate for the accurate identification of the bacterial subtypes of interest in a specimen. This applied model for microbiology laboratory use is based on a Bayesian mode of inference, which combines two inputs: (ii) a prespecified estimate, or prior distribution statement, based on available scientific knowledge and (ii) observed data. The specific inputs for the model are a prior distribution statement of the number of strains per specimen provided by an informed microbiologist and data from a microbiological survey indicating the number of strains per specimen. The model output is an updated probability distribution of strains per specimen, which can be used to estimate the probability of observing all strains present according to the number of colonies that are sampled. In this report two scenarios that illustrate the use of the model to estimate bacterial colony sample size requirements are presented. In the first scenario, bacterial colony sample size is estimated to correctly identify Campylobacter amplified restriction fragment length polymorphism types on broiler carcasses. The second scenario estimates bacterial colony sample size to correctly identify Salmonella enterica serotype Enteritidis phage types in fecal drag swabs from egg-laying poultry flocks. An advantage of the model is that as updated inputs from ongoing surveys are incorporated into the model, increasingly precise sample size estimates are likely to be made. PMID- 14532079 TI - Cloning and characterization of the xyl1 gene, encoding an NADH-preferring xylose reductase from Candida parapsilosis, and its functional expression in Candida tropicalis. AB - Xylose reductase (XR) is a key enzyme in D-xylose metabolism, catalyzing the reduction of D-xylose to xylitol. An NADH-preferring XR was purified to homogeneity from Candida parapsilosis KFCC-10875, and the xyl1 gene encoding a 324-amino-acid polypeptide with a molecular mass of 36,629 Da was subsequently isolated using internal amino acid sequences and 5' and 3' rapid amplification of cDNA ends. The C. parapsilosis XR showed high catalytic efficiency (kcat/Km = 1.46 s(-1) mM(-1)) for D-xylose and showed unusual coenzyme specificity, with greater catalytic efficiency with NADH (kcat/Km = 1.39 x 10(4) s(-1) mM(-1)) than with NADPH (kcat/Km = 1.27 x 10(2) s(-1) mM(-1)), unlike all other aldose reductases characterized. Studies of initial velocity and product inhibition suggest that the reaction proceeds via a sequentially ordered Bi Bi mechanism, which is typical of XRs. Candida tropicalis KFCC-10960 has been reported to have the highest xylitol production yield and rate. It has been suggested, however, that NADPH-dependent XRs, including the XR of C. tropicalis, are limited by the coenzyme availability and thus limit the production of xylitol. The C. parapsilosis xyl1 gene was placed under the control of an alcohol dehydrogenase promoter and integrated into the genome of C. tropicalis. The resulting recombinant yeast, C. tropicalis BN-1, showed higher yield and productivity (by 5 and 25%, respectively) than the wild strain and lower production of by-products, thus facilitating the purification process. The XRs partially purified from C. tropicalis BN-1 exhibited dual coenzyme specificity for both NADH and NADPH, indicating the functional expression of the C. parapsilosis xyl1 gene in C. tropicalis BN-1. This is the first report of the cloning of an xyl1 gene encoding an NADH-preferring XR and its functional expression in C. tropicalis, a yeast currently used for industrial production of xylitol. PMID- 14532081 TI - Assessment of the effects of holding time and temperature on Escherichia coli densities in surface water samples. AB - Escherichia coli is a routinely used microbiological indicator of water quality. To determine whether holding time and storage conditions had an effect on E. coli densities in surface water, studies were conducted in three phases, encompassing 24 sites across the United States and four commonly used monitoring methods. During all three phases of the study, E. coli samples were analyzed at time 0 and at 8, 24, 30, and 48 h after sample collection. During phase 1, when 4 degrees C samples were evaluated by Colilert or by placing a membrane onto mFC medium followed by transfer to nutrient agar containing 4-methylumbelliferyl-beta-D glucuronide (mFC/NA-MUG), three of four sites showed no significant differences throughout the 48-h study. During phase 2, five of seven sites showed no significant difference between time 0 and 24 h by membrane filtration (mFC/NA MUG). When evaluated by the Colilert method, five of seven sites showed no significant difference in E. coli density between time 0 and 48 h. During phase 3, 8 of 13 sites showed no significant differences in E. coli densities between time 0 and the 48-h holding time, regardless of method. Based on the results of these studies, it appears that if samples are held below 10 degrees C and are not allowed to freeze, most surface water E. coli samples analyzed by commonly used methods beyond 8 h after sample collection can generate E. coli data comparable to those generated within 8 h of sample collection. Notwithstanding this conclusion, E. coli samples collected from surface waters should always be analyzed as soon as possible. PMID- 14532080 TI - Statistical approaches for estimating actinobacterial diversity in marine sediments. AB - Bacterial diversity in a deep-sea sediment was investigated by constructing actinobacterium-specific 16S ribosomal DNA (rDNA) clone libraries from sediment sections taken 5 to 12, 15 to 18, and 43 to 46 cm below the sea floor at a depth of 3,814 m. Clones were placed into operational taxonomic unit (OTU) groups with >/= 99% 16S rDNA sequence similarity; the cutoff value for an OTU was derived by comparing 16S rRNA homology with DNA-DNA reassociation values for members of the class Actinobacteria. Diversity statistics were used to determine how the level of dominance, species richness, and genetic diversity varied with sediment depth. The reciprocal of Simpson's index (1/D) indicated that the pattern of diversity shifted toward dominance from uniformity with increasing sediment depth. Nonparametric estimation of the species richness in the 5- to 12-, 15- to 18-, and 43- to 46-cm sediment sections revealed a trend of decreasing species number with depth, 1,406, 308, and 212 OTUs, respectively. Application of the LIBSHUFF program indicated that the 5- to 12-cm clone library was composed of OTUs significantly (P = 0.001) different from those of the 15- to 18- and 43- to 46-cm libraries. F(ST) and phylogenetic grouping of taxa (P tests) were both significant (P < 0.00001 and P < 0.001, respectively), indicating that genetic diversity decreased with sediment depth and that each sediment community harbored unique phylogenetic lineages. It was also shown that even nonconservative OTU definitions result in severe underestimation of species richness; unique phylogenetic clades detected in one OTU group suggest that OTUs do not correspond to real ecological groups sensu Palys (T. Palys, L. K. Nakamura, and F. M. Cohan, Int. J. Syst. Bacteriol. 47:1145-1156, 1997). Mechanisms responsible for diversity and their implications are discussed. PMID- 14532082 TI - Use of bromodeoxyuridine immunocapture to identify active bacteria associated with arbuscular mycorrhizal hyphae. AB - Arbuscular mycorrhizae are beneficial for crops grown under low-till management systems. Increasingly, it is becoming apparent that bacteria associated with mycorrhizae can enhance the beneficial relationship between mycorrhizae and plants. However, it has been difficult to study these relationships by conventional techniques. In this study actively growing bacteria were identified in soil from an undisturbed fallow field known to contain arbuscular mycorrhizae by using molecular tools to eliminate the need for cultivation. A thymidine analog, bromodeoxyuridine (BrdU), was added to the soil and incubated for 2 days. DNA was extracted, and the newly synthesized DNA was isolated by immunocapture of the BrdU-containing DNA. The active bacteria in the community were identified by 16S rRNA gene PCR amplification and DNA sequence analysis. Based on 16S rRNA gene sequence information, a selective medium was chosen to isolate the corresponding active bacteria. Bacillus cereus strain VA1, one of the bacteria identified by the BrdU method, was isolated from the soil and tagged with green fluorescent protein. By using confocal microscopy, this bacterium was shown to clearly attach to arbuscular mycorrhizal hyphae. This study was the first to use this combination of molecular and traditional approaches to isolate, identify, and visualize a specific bacterium that is active in fallow soil and associates with arbuscular mycorrhizal hyphae. PMID- 14532083 TI - Nuclease activity of Saccharomyces cerevisiae Mre11 functions in targeted nucleotide alteration. AB - Oligonucleotides can be used to direct site-specific changes in genomic DNA through a process in which mismatched base pairs in the oligonucleotide and the target DNA are created. The mechanism by which these complexes are developed and resolved is being studied by using Saccharomyces cerevisiae as a model system. Genetic analyses have revealed that in all likelihood the reaction occurs in two phases: DNA pairing and DNA repair. While the former phase involves strand assimilation, the latter phase likely involves an endonucleolytic processing step that leads to joint resolution. In this study, we established the importance of a functioning MRE11 gene in the overall reaction, as yeast strains deficient in MRE11 exhibited severely reduced activity. The activity could be rescued by complementation with wild-type MRE11 genes but not with MRE11 alleles lacking the nuclease function. Taken together, the data suggest that Mre11 provides nuclease activity for targeted nucleotide exchange, a process that could be used to reengineer yeast genes. PMID- 14532084 TI - Spiralin is not essential for helicity, motility, or pathogenicity but is required for efficient transmission of Spiroplasma citri by its leafhopper vector Circulifer haematoceps. AB - Spiralin is the most abundant protein at the surface of the plant pathogenic mollicute Spiroplasma citri and hence might play a role in the interactions of the spiroplasma with its host plant and/or its insect vector. To study spiralin function, mutants were produced by inactivating the spiralin gene through homologous recombination. A spiralin-green fluorescent protein (GFP) translational fusion was engineered and introduced into S. citri by using an oriC based targeting vector. According to the strategy used, integration of the plasmid by a single-crossover recombination at the spiralin gene resulted in the expression of the spiralin-GFP fusion protein. Two distinct mutants were isolated. Western and colony immunoblot analyses showed that one mutant (GII3 9a5) did produce the spiralin-GFP fusion protein, which was found not to fluoresce, whereas the other (GII3-9a2) produced neither the fusion protein nor the wild-type spiralin. Both mutants displayed helical morphology and motility, similarly to the wild-type strain GII-3. Genomic DNA analyses revealed that GII3 9a5 was unstable and that GII3-9a2 was probably derived from GII3-9a5 by a double crossover recombination between plasmid sequences integrated into the GII3-9a5 chromosome and free plasmid. When injected into the leafhopper vector Circulifer haematoceps, the spiralinless mutant GII3-9a2 multiplied to high titers in the insects (1.1 x 10(6) to 2.8 x 10(6) CFU/insect) but was transmitted to the host plant 100 times less efficiently than the wild-type strain. As a result, not all plants were infected, and symptom production in these plants was delayed for 2 to 4 weeks compared to that in the wild-type strain. In the infected plants however, the mutant multiplied to high titers (1.2 x 10(6) to 1.4 x 10(7) CFU/g of midribs) and produced the typical symptoms of the disease. These results indicate that spiralin is not essential for pathogenicity but is required for efficient transmission of S. citri by its insect vector. PMID- 14532085 TI - Prospecting for novel biocatalysts in a soil metagenome. AB - The metagenomes of complex microbial communities are rich sources of novel biocatalysts. We exploited the metagenome of a mixed microbial population for isolation of more than 15 different genes encoding novel biocatalysts by using a combined cultivation and direct cloning strategy. A 16S rRNA sequence analysis revealed the presence of hitherto uncultured microbes closely related to the genera Pseudomonas, Agrobacterium, Xanthomonas, Microbulbifer, and Janthinobacterium. Total genomic DNA from this bacterial community was used to construct cosmid DNA libraries, which were functionally searched for novel enzymes of biotechnological value. Our searches in combination with cosmid sequencing resulted in identification of four clones encoding 12 putative agarase genes, most of which were organized in clusters consisting of two or three genes. Interestingly, nine of these agarase genes probably originated from gene duplications. Furthermore, we identified by DNA sequencing several other biocatalyst-encoding genes, including genes encoding a putative stereoselective amidase (amiA), two cellulases (gnuB and uvs080), an alpha-amylase (amyA), a 1,4 alpha-glucan branching enzyme (amyB), and two pectate lyases (pelA and uvs119). Also, a conserved cluster of two lipase genes was identified, which was linked to genes encoding a type I secretion system. The novel gene aguB was overexpressed in Escherichia coli, and the enzyme activities were determined. Finally, we describe more than 162 kb of DNA sequence that provides a strong platform for further characterization of this microbial consortium. PMID- 14532086 TI - Detection of Prochlorothrix in brackish waters by specific amplification of pcb genes. AB - Prochlorothrix hollandica is the only filamentous chlorophyll b (Chlb)-containing oxyphotobacterium that has been found in freshwater habitats to date. Chlb serves as a light-harvesting pigment which is bound to special binding proteins (Pcb). Even though Prochlorothrix was initially characterized as a highly salt-sensitive species, we detected it in a brackish water environment that is characterized by salinities of up to 12 practical salinity units. Using PCR and reverse transcription, we amplified pcb gene fragments of phytoplankton samples taken along a salinity gradient in the eutrophic Darss-Zingst estuary (southern Baltic Sea). After sequencing, high levels of homology to the pcbB and pcbC genes of P. hollandica were found. Furthermore, autofluorescence of Prochlorothrix-like filaments that indicated that Chlb was present was detected in enrichment cultures prepared from the estuarine phytoplankton. The detection of Chlb containing filaments, as well as the pcb and 16S ribosomal DNA sequences, suggests that Prochlorothrix is an indigenous genus in the Darss-Zingst estuary and may also inhabit many other brackish water environments. The potential of using pcb gene detection to differentiate Prochlorothrix from morphologically indistinguishable species belonging to the genera Pseudanabaena and Planktothrix (Oscillatoria) in phytoplankton analyses is discussed. PMID- 14532087 TI - Invasion of spores of the arbuscular mycorrhizal fungus Gigaspora decipiens by Burkholderia spp. AB - Burkholderia species are bacterial soil inhabitants that are capable of interacting with a variety of eukaryotes, in some cases occupying intracellular habitats. Pathogenic and nonpathogenic Burkholderia spp., including B. vietnamiensis, B. cepacia, and B. pseudomallei, were grown on germinating spores of the arbuscular mycorrhizal fungus Gigaspora decipiens. Spore lysis assays revealed that all Burkholderia spp. tested were able to colonize the interior of G. decipiens spores. Amplification of specific DNA sequences and transmission electron microscopy confirmed the intracellular presence of B. vietnamiensis. Twelve percent of all spores were invaded by B. vietnamiensis, with an average of 1.5 x 10(6) CFU recovered from individual infected spores. Of those spores inoculated with B. pseudomallei, 7% were invaded, with an average of 5.5 x 10(5) CFU recovered from individual infected spores. Scanning electron and fluorescence microscopy provided insights into the morphology of surfaces of spores and hyphae of G. decipiens and the attachment of bacteria. Burkholderia spp. colonized both hyphae and spores, attaching to surfaces in either an end-on or side-on fashion. Adherence of Burkholderia spp. to eukaryotic surfaces also involved the formation of numerous fibrillar structures. PMID- 14532088 TI - A novel extracellular multicopper oxidase from Phanerochaete chrysosporium with ferroxidase activity. AB - Lignin degradation by the white rot basidiomycete Phanerochaete chrysosporium involves various extracellular oxidative enzymes, including lignin peroxidase, manganese peroxidase, and a peroxide-generating enzyme, glyoxal oxidase. Recent studies have suggested that laccases also may be produced by this fungus, but these conclusions have been controversial. We identified four sequences related to laccases and ferroxidases (Fet3) in a search of the publicly available P. chrysosporium database. One gene, designated mco1, has a typical eukaryotic secretion signal and is transcribed in defined media and in colonized wood. Structural analysis and multiple alignments identified residues common to laccase and Fet3 sequences. A recombinant MCO1 (rMCO1) protein expressed in Aspergillus nidulans had a molecular mass of 78 kDa, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and the copper I-type center was confirmed by the UV-visible spectrum. rMCO1 oxidized various compounds, including 2,2' azino(bis-3-ethylbenzthiazoline-6-sulfonate) (ABTS) and aromatic amines, although phenolic compounds were poor substrates. The best substrate was Fe2+, with a Km close to 2 micro M. Collectively, these results suggest that the P. chrysosporium genome does not encode a typical laccase but rather encodes a unique extracellular multicopper oxidase with strong ferroxidase activity. PMID- 14532089 TI - Detection of the free-living forms of sulfide-oxidizing gill endosymbionts in the lucinid habitat (Thalassia testudinum environment). AB - Target DNA from the uncultivable Codakia orbicularis endosymbiont was PCR amplified from sea-grass sediment. To confirm that such amplifications originated from intact bacterial cells rather than free DNA, whole-cell hybridization (fluorescence in situ hybridization technique) with the specific probe Symco2 was performed along with experimental infection of aposymbiotic juveniles placed in contact with the same sediment. Taken together, the data demonstrate that the sulfide-oxidizing gill endosymbiont of Codakia orbicularis is present in the environment as a free-living uncultivable form. PMID- 14532090 TI - An alternative efficient procedure for purification of the obligate intracellular fish bacterial pathogen Piscirickettsia salmonis. AB - Piscirickettsia salmonis is an obligate intracellular bacterial pathogen of salmonid fish and the etiological agent of the aggressive disease salmonid rickettsial syndrome. Today, this disease, also known as piscirickettsiosis, is the cause of high mortality in net pen-reared salmonids in southern Chile. Although the bacteria can be grown in tissue culture cells, genetic analysis of the organism has been hindered because of the difficulty in obtaining P. salmonis DNA free from contaminating host cell DNA. In this report, we describe a novel procedure to purify in vitro-grown bacteria with iodixanol as the substrate to run differential centrifugation gradients which, combined with DNase I digestion, yield enough pure bacteria to do DNA analysis. The efficiency of the purification procedure relies on two main issues: semiquantitative synchrony of the P. salmonis-infected Chinook salmon embryo (CHSE-214) tissue culture cells and low osmolarity of iodixanol to better resolve bacteria from the membranous structures of the host cell. This method resulted in the isolation of intact piscirickettsia organisms and removed salmon and mitochondrial DNA effectively, with only 1.0% contamination with the latter. PMID- 14532091 TI - Identification of peptides that mimic the pertussis toxin binding site on bovine fetuin. AB - The introduction of acellular pertussis vaccines has greatly enhanced the safety profile of vaccines to prevent whooping cough. Pertussis toxin (Ptx) is one component produced by Bordetella pertussis that is contained in all of these vaccines, either in combination with other known pertussis virulence factors or as the sole pertussis component, combined with tetanus and diphtheria toxoids. A hydrogen peroxide toxoid of Ptx has been shown to be efficacious in preventing pertussis infections in a mass vaccination trial and is presently licensed in the United States and Europe (B. Trollfors, J. Taranger, T. Lagergard, L. Lind, V. Sundh, G. Zackrisson, C. U. Lowe, W. Blackwelder, and J. B. Robbins, N. Engl. J. Med. 333:1045-1050, 1995). The industrial production of Ptx can be performed through the cultivation of B. pertussis in well-defined growth media, in which the components can be well characterized and their origins can be documented. Once the bacteria are removed from the culture, Ptx can be isolated from the supernatant and purified by using the technique described by Sekura et al. (R. D. Sekura, F. Fish, C. R. Manclark, B. Meade, and Y. L. Zhang, J. Biol. Chem. 258:14647-14651, 1983). The only drawback of this procedure, which combines two affinity chromatography steps, one with Blue Sepharose and a second with matrix bound bovine fetuin (BF), is the source and purity of the BF. Concern about vaccine preparations that may possibly risk contamination by material associated with bovine spongioform encephalopathy has continued to increase. We thus sought a replacement for the BF affinity chromatography and, more specifically, for the glycosidic moiety on BF. We describe here the identification of a seven-amino acid peptide that mimics the glycosidic moiety on BF to which Ptx binds. Furthermore, we have constructed an affinity column containing this peptide that can be used to replace BF in Ptx purification. Finally, we used the X-ray crystallographic structure of Ptx bound to the oligosaccharide moiety of BF as a scaffold and replaced the oligosaccharide with the peptide. PMID- 14532092 TI - Comparison of velocity profiles for different flow chamber designs used in studies of microbial adhesion to surfaces. AB - Flow chambers are commonly used to study microbial adhesion to surfaces under environmentally relevant hydrodynamic conditions. The parallel plate flow chamber (PPFC) is the most common design, and mass transport occurs through slow convective diffusion. In this study, we analyzed four different PPFCs to determine whether the expected hydrodynamic conditions, which control both mass transport and detachment forces, are actually achieved. Furthermore, the different PPFCs were critically evaluated based on the size of the area where the velocity profile was established and constant with a range of flow rates, indicating that valid observations could be made. Velocity profiles in the different chambers were calculated by using a numerical simulation model based on the finite element method and were found to coincide with the profiles measured by particle image velocimetry. Environmentally relevant shear rates between 0 and 10,000 s(-1) could be measured over a sizeable proportion of the substratum surface for only two of the four PPFCs. Two models appeared to be flawed in the design of their inlets and outlets and allowed development of a stable velocity profile only for shear rates up to 0.5 and 500 s(-1). For these PPFCs the inlet and outlet were curved, and the modeled shear rates deviated from the calculated shear rates by up to 75%. We concluded that PPFCs used for studies of microbial adhesion to surfaces should be designed so that their inlets and outlets are in line with the flow channel. Alternatively, the channel length should be increased to allow a greater length for the establishment of the desired hydrodynamic conditions. PMID- 14532094 TI - Single-cell enumeration of an uncultivated TM7 subgroup in the human subgingival crevice. AB - Specific oligonucleotide hybridization conditions were established for single cell enumeration of uncultivated TM7 and IO25 bacteria by using clones expressing heterologous 16S rRNA. In situ analysis of human subgingival crevice specimens revealed that a greater proportion of samples from sites of chronic periodontitis than from healthy sites contained TM7 subgroup IO25. In addition, IO25 bacterial cells from periodontitis site samples were more abundant and fourfold longer than IO25 cells from healthy site samples. PMID- 14532093 TI - Species-specific peptide ligands for the detection of Bacillus anthracis spores. AB - Currently available detectors for spores of Bacillus anthracis, the causative agent of anthrax, are inadequate for frontline use and general monitoring. There is a critical need for simple, rugged, and inexpensive detectors capable of accurate and direct identification of B. anthracis spores. Necessary components in such detectors are stable ligands that bind tightly and specifically to target spores. By screening a phage display peptide library, we identified a family of peptides, with the consensus sequence TYPXPXR, that bind selectively to B. anthracis spores. We extended this work by identifying a peptide variant, ATYPLPIR, with enhanced ability to bind to B. anthracis spores and an additional peptide, SLLPGLP, that preferentially binds to spores of species phylogenetically similar to, but distinct from, B. anthracis. These two peptides were used in tandem in simple assays to rapidly and unambiguously identify B. anthracis spores. We envision that these peptides can be used as sensors in economical and portable B. anthracis spore detectors that are essentially free of false-positive signals due to other environmental Bacillus spores. PMID- 14532095 TI - Development of a mariner-based transposon for use in Sorangium cellulosum. AB - In order to generate marked insertions in the myxobacterium Sorangium cellulosum, a transposon based on the eukaryotic mariner transposon was developed. The transposition frequency was increased with the use of a mutated tnp gene. The transposon randomly inserts into the chromosome, as demonstrated by targeted mutagenesis of the epoK gene. PMID- 14532097 TI - Effects of minerals on resistance of Bacillus subtilis spores to heat and hydrostatic pressure. AB - Among Bacillus subtilis IFO13722 spores sporulated at 30, 37, and 44 degrees C, those sporulated at 30 degrees C had the highest resistance to treatments with high hydrostatic pressure (100 to 300 MPa, 55 degrees C, 30 min). Pressure resistance increased after demineralization of the spores and decreased after remineralization of the spores with Ca(2+) or Mg(2+), whereas the resistance did not change when spores were remineralized with Mn(2+) or K(+), suggesting that former two divalent ions were involved in the activation of cortex-lytic enzymes during germination. PMID- 14532096 TI - Application of host-specific bacteriophages to the surface of chicken skin leads to a reduction in recovery of Campylobacter jejuni. AB - Retail poultry products are widely purported as the major infection vehicle for human campylobacteriosis. Numerous intervention strategies have sought to reduce Campylobacter contamination on broiler carcasses in the abattoir. This study reports the efficacy of bacteriophage in reducing the number of recoverable Campylobacter jejuni cells on artificially contaminated chicken skin. PMID- 14532098 TI - Teschoviruses as indicators of porcine fecal contamination of surface water. AB - Teschoviruses specifically infect pigs and are shed in pig feces. Hence, their presence in water should indicate contamination with pig fecal residues. To assess this hypothesis, we have developed a real-time reverse transcriptase PCR (RT-PCR) method that allows the quantitative detection of pig teschovirus (PTV) RNA. The method is able to detect 92 fg of PTV RNA per ml of sample. Using this method, we have detected the presence of PTV RNA in water and fecal samples from all pig farms examined (n = 5). Feces from other animal species (cattle, sheep, and goats) were negative in this test. To compare the PTV RNA detection method with conventional chemical determinations currently in use for evaluation of water contamination, we analyzed water samples collected downstream from a pig slurry spillage site. We have found a positive correlation within both types of determinations. The sensitivity of the PTV detection assay was similar to that achieved by unspecific organic matter determination and superior to all other conventional chemical analyses performed. Furthermore, the new method is highly specific, revealing the porcine origin of the contamination, a feature that is lacking in currently available methods for the assessment of water contamination. PMID- 14532099 TI - Comparison of Campylobacter isolates from poultry and humans: association between in vitro virulence properties, biotypes, and pulsed-field gel electrophoresis clusters. AB - The in vitro virulence properties of 197 temporally and geographically related Campylobacter isolates from chicken broilers and humans were compared. Comparisons of the virulence properties associated with genotypes and biotypes were made. All isolates adhered to, and 63% invaded, INT-407 cells, whereas 13% were cytotoxic for CHO cells. CHO cell-cytotoxic extracts were also cytotoxic for INT-407 cells, but the sensitivity for Vero cells was variable. The proportion of isolates demonstrating a high invasiveness potential (>1,000 CFU ml(-1)) or Vero cell cytotoxicity was significantly higher for human than for poultry isolates. Invasiveness was associated with Campylobacter jejuni isolates of biotypes 1 and 2, whereas CHO and INT-407 cell cytotoxicity was associated with C. jejuni isolates of biotypes 3 and 4. Cytotoxic isolates were also clustered according to pulsed-field gel electrophoresis profiles. PMID- 14532100 TI - Formate-dependent growth and homoacetogenic fermentation by a bacterium from human feces: description of Bryantella formatexigens gen. nov., sp. nov. AB - Formate stimulates growth of a new bacterium from human feces. With high formate, it ferments glucose to acetate via the Wood-Ljungdahl pathway. The original isolate fermented vegetable cellulose and carboxymethylcellulose, but it lost this ability after storage at -76 degrees C. 16S rRNA gene sequencing identifies it as a distinct line within the Clostridium coccoides supra-generic rRNA grouping. We propose naming it Bryantella formatexigens gen. nov., sp. nov. PMID- 14532101 TI - Multiplex real-time PCR method to identify Shiga toxin genes stx1 and stx2 and Escherichia coli O157:H7/H- serotype. AB - A multiplex real-time PCR method to simultaneously detect the stx1 and stx2 genes of Shiga toxin-producing Escherichia coli and a unique conserved single nucleotide polymorphism in the E. coli O157:H7/H- uidA gene has been developed. There is more than 98.6% sensitivity and 100% specificity for all three gene targets based on a panel of 138 isolates. The PCR efficiencies were >/= 1.89, and as few as 6 CFU/reaction could be detected. PMID- 14532102 TI - The high salt requirement of the moderate halophile Chromohalobacter salexigens DSM3043 can be met not only by NaCl but by other ions. AB - The growth rate of Chromohalobacter salexigens DSM 3043 can be stimulated in media containing 0.3 M NaCl by a 0.7 M concentration of other salts of Na+, K+, Rb+, or NH4+, Cl-, Br-, NO3-, or SO4(2-) ions. To our knowledge, growth rate stimulation by a general high ion concentration has not been reported for any organism previously. PMID- 14532103 TI - Petroleum pollution bioremediation using water-insoluble uric acid as the nitrogen source. AB - The biodegradation of hydrocarbon pollutants in open systems is limited by the availability of a utilizable nitrogen source. This limitation can be overcome by using uric acid. Enrichment cultures grown on crude oil-uric acid media yielded mixed and pure cultures that degraded petroleum. In a simulated open system, uric acid bound to crude oil and was available for bacterial growth and petroleum biodegradation. PMID- 14532104 TI - Bacterial activity in South pole snow is questionable. PMID- 14532106 TI - EZH2 is downstream of the pRB-E2F pathway, essential for proliferation and amplified in cancer. AB - Recent experiments have demonstrated that the Polycomb group (PcG) gene EZH2 is highly expressed in metastatic prostate cancer and in lymphomas. EZH2 is a component of the PRC2 histone methyltransferase complex, which also contains EED and SUZ12 and is required for the silencing of HOX gene expression during embryonic development. Here we demonstrate that both EZH2 and EED are essential for the proliferation of both transformed and non-transformed human cells. In addition, the pRB-E2F pathway tightly regulates their expression and, consistent with this, we find that EZH2 is highly expressed in a large set of human tumors. These results raise the question whether EZH2 is a marker of proliferation or if it is actually contributing to tumor formation. Significantly, we propose that EZH2 is a bona fide oncogene, since we find that ectopic expression of EZH2 is capable of providing a proliferative advantage to primary cells and, in addition, its gene locus is specifically amplified in several primary tumors. PMID- 14532107 TI - Novel roles for the flagellum in cell morphogenesis and cytokinesis of trypanosomes. AB - Flagella and cilia are elaborate cytoskeletal structures conserved from protists to mammals, where they fulfil functions related to motility or sensitivity. Here we demonstrate novel roles for the flagellum in the control of cell size, shape, polarity and division of the protozoan Trypanosoma brucei. To investigate the function of the flagellum, its formation was perturbed by inducible RNA interference silencing of com ponents required for intraflagellar transport, a dynamic process necessary for flagellum assembly. First, we show that down regulation of intraflagellar transport leads to assembly of a shorter flagellum. Strikingly, cells with a shorter flagellum are smaller, with a direct correlation between flagellum length and cell size. Detailed morphogenetic analysis reveals that the tip of the new flagellum defines the point where cytokinesis is initiated. Secondly, when new flagellum formation is completely blocked, non flagellated cells are very short, lose their normal shape and polarity, and fail to undergo cytokinesis. We show that flagellum elongation controls formation of cytoskeletal structures (present in the cell body) that act as molecular organizers of the cell. PMID- 14532108 TI - Synaptotagmin 7 splice variants differentially regulate synaptic vesicle recycling. AB - The speed of synaptic vesicle recycling determines the efficacy of neurotransmission during repetitive stimulation. Synaptotagmins are synaptic C(2) domain proteins that are involved in exocytosis, but have also been linked to endocytosis. We now demonstrate that upon expression in transfected neurons, a short splice variant of synaptotagmin 7 that lacks C(2)-domains accelerates endocytic recycling of synaptic vesicles, whereas a longer splice variant that contains C(2)-domains decelerates recycling. These results suggest that alternative splicing of synaptotagmin 7 acts as a molecular switch, which targets vesicles to fast and slow recycling pathways. PMID- 14532109 TI - Structural basis for Nup2p function in cargo release and karyopherin recycling in nuclear import. AB - The yeast nucleoporin Nup2p is associated primarily with the nuclear basket of nuclear pore complexes and is required for efficient importin-alpha:beta-mediated nuclear protein import as well as efficient nuclear export of Kap60p/importin alpha. Residues 1-51 of Nup2p bind tightly to Kap60p and are required for Nup2p function in vivo. We have determined the 2.6 A resolution crystal structure of a complex between this region of Nup2p and the armadillo repeat domain of Kap60p. Nup2p binds along the inner concave groove of Kap60p, but its interaction interface is different from that employed for nuclear localization signal (NLS) recognition although there is some overlap between them. Nup2p binds Kap60p more strongly than NLSs and accelerates release of NLSs from Kap60p. Nup2p itself is released from Kap60p by Cse1p:RanGTP only in the presence of the importin-beta binding (IBB) domain of Kap60p. These data indicate that Nup2p increases the overall rate of nuclear trafficking by coordinating nuclear import termination and importin recycling as a concerted process. PMID- 14532110 TI - Mitochondrial translocation contact sites: separation of dynamic and stabilizing elements in formation of a TOM-TIM-preprotein supercomplex. AB - Preproteins with N-terminal presequences are imported into mitochondria at translocation contact sites that include the translocase of the outer membrane (TOM complex) and the presequence translocase of the inner membrane (TIM23 complex). Little is known about the functional cooperation of these translocases. We have characterized translocation contact sites by a productive TOM-TIM preprotein supercomplex to address the role of three translocase subunits that expose domains to the intermembrane space (IMS). The IMS domain of the receptor Tom22 is required for stabilization of the translocation contact site supercomplex. Surprisingly, the N-terminal segment of the channel Tim23, which tethers the TIM23 complex to the outer membrane, is dispensable for both protein import and generation of the TOM-TIM supercomplex. Tim50, with its large IMS domain, is crucial for generation but not for stabilization of the supercomplex. Thus, Tim50 functions as a dynamic factor and the IMS domain of Tom22 represents a stabilizing element in formation of a productive translocation contact site supercomplex. PMID- 14532111 TI - Rotation of the stalk/neck and one head in a new crystal structure of the kinesin motor protein, Ncd. AB - Molecular motors undergo conformational changes to produce force and move along cytoskeletal filaments. Structural changes have been detected in kinesin motors; however, further changes are expected because previous crystal structures are in the same or closely related conformations. We report here a 2.5 A crystal structure of the minus-end kinesin, Ncd, with the coiled-coil stalk/neck and one head rotated by approximately 75 degrees relative to the other head. The two heads are asymmetrically positioned with respect to the stalk and show asymmetry of nucleotide state: one head is fully occupied, but the other is unstably bound to ADP. Unlike previous structures, our new atomic model can be fit into cryoelectron microscopy density maps of the motor attached to microtubules, where it appears to resemble a one-head-bound motor with the stalk rotated towards the minus end. Interactions between neck and motor core residues, observed in the head that moves with the stalk, are disrupted in the other head, permitting rotation of the stalk/neck. The rotation could represent a force-producing stroke that directs the motor to the minus end. PMID- 14532112 TI - Interaction of fascin and protein kinase Calpha: a novel intersection in cell adhesion and motility. AB - Coordination of protrusive and contractile cell-matrix contacts is important for cell adhesion and migration, but the mechanisms involved are not well understood. We report an unexpected direct association between fascin, an actin-bundling component of filopodia, microspikes and lamellipodial ribs, and protein kinase Calpha (PKCalpha), a regulator of focal adhesions. The association is detectable by protein-protein binding in vitro, by coimmunoprecipitation from cell extracts, and in live cells as fluorescence resonance energy transfer detected by fluorescence imaging lifetime microscopy. The interaction is physiologically regulated by the extracellular matrix context of cells, depends on activation of PKCalpha and is mediated by the C1B domain of PKCalpha. Strikingly, a fascin mutant, fascin S39D, associates constitutively with PKCalpha. Through use of a newly developed set of membrane-permeable peptides that separately inhibit either fascin/PKCalpha or fascin/actin binding, we have uncovered that specific blockade of the fascin/PKCalpha interaction increases cell migration on fibronectin in conjunction with increased fascin protrusions and remodeling of focal adhesions. These results identify the fascin-PKCalpha interaction as an important novel intersection in the regulation and networking of cell-matrix contacts. PMID- 14532113 TI - Mechanisms underlying excitatory effects of group I metabotropic glutamate receptors via inhibition of 2P domain K+ channels. AB - Group I metabotropic glutamate receptors (mGluRs) are implicated in diverse processes such as learning, memory, epilepsy, pain and neuronal death. By inhibiting background K(+) channels, group I mGluRs mediate slow and long-lasting excitation. The main neuronal representatives of this K(+) channel family (K(2P) or KCNK) are TASK and TREK. Here, we show that in cerebellar granule cells and in heterologous expression systems, activation of group I mGluRs inhibits TASK and TREK channels. D-myo-inositol-1,4,5-triphosphate and phosphatidyl-4,5-inositol biphosphate depletion are involved in TASK channel inhibition, whereas diacylglycerols and phosphatidic acids directly inhibit TREK channels. Mechanisms described here with group I mGluRs will also probably stand for many other receptors of hormones and neurotransmitters. PMID- 14532114 TI - Phosphatidylinositol-4,5-bisphosphate, PIP2, controls KCNQ1/KCNE1 voltage-gated potassium channels: a functional homology between voltage-gated and inward rectifier K+ channels. AB - Phosphatidylinositol-4,5-bisphosphate (PIP(2)) is a major signaling molecule implicated in the regulation of various ion transporters and channels. Here we show that PIP(2) and intracellular MgATP control the activity of the KCNQ1/KCNE1 potassium channel complex. In excised patch-clamp recordings, the KCNQ1/KCNE1 current decreased spontaneously with time. This rundown was markedly slowed by cytosolic application of PIP(2) and fully prevented by application of PIP(2) plus MgATP. PIP(2)-dependent rundown was accompanied by acceleration in the current deactivation kinetics, whereas the MgATP-dependent rundown was not. Cytosolic application of PIP(2) slowed deactivation kinetics and also shifted the voltage dependency of the channel activation toward negative potentials. Complex changes in the current characteristics induced by membrane PIP(2) was fully restituted by a model originally elaborated for ATP-regulated two transmembrane-domain potassium channels. The model is consistent with stabilization by PIP(2) of KCNQ1/KCNE1 channels in the open state. Our data suggest a striking functional homology between a six transmembrane-domain voltage-gated channel and a two transmembrane-domain ATP-gated channel. PMID- 14532115 TI - Loss of K-Cl co-transporter KCC3 causes deafness, neurodegeneration and reduced seizure threshold. AB - K-Cl co-transporters are encoded by four homologous genes and may have roles in transepithelial transport and in the regulation of cell volume and cytoplasmic chloride. KCC3, an isoform mutated in the human Anderman syndrome, is expressed in brain, epithelia and other tissues. To investigate the physiological functions of KCC3, we disrupted its gene in mice. This severely impaired cell volume regulation as assessed in renal tubules and neurons, and moderately raised intraneuronal Cl(-) concentration. Kcc3(-/-) mice showed severe motor abnormalities correlating with a progressive neurodegeneration in the peripheral and CNS. Although no spontaneous seizures were observed, Kcc3(-/-) mice displayed reduced seizure threshold and spike-wave complexes on electrocorticograms. These resembled EEG abnormalities in patients with Anderman syndrome. Kcc3(-/-) mice also displayed arterial hypertension and a slowly progressive deafness. KCC3 was expressed in many, but not all cells of the inner ear K(+) recycling pathway. These cells slowly degenerated, as did sensory hair cells. The present mouse model has revealed important cellular and systemic functions of KCC3 and is highly relevant for Anderman syndrome. PMID- 14532116 TI - Caspase-12 and endoplasmic reticulum stress mediate neurotoxicity of pathological prion protein. AB - Prion diseases are characterized by accumulation of misfolded prion protein (PrP(Sc)), and neuronal death by apoptosis. Here we show that nanomolar concentrations of purified PrP(Sc) from mouse scrapie brain induce apoptosis of N2A neuroblastoma cells. PrP(Sc) toxicity was associated with an increase of intracellular calcium released from endoplasmic reticulum (ER) and up-regulation of several ER chaperones. Caspase-12 activation was detected in cells treated with PrP(Sc), and cellular death was inhibited by overexpression of a catalytic mutant of caspase-12 or an ER-targeted Bcl-2 chimeric protein. Scrapie-infected N2A cells were more susceptible to ER-stress and to PrP(Sc) toxicity than non infected cells. In scrapie-infected mice a correlation between caspase-12 activation and neuronal loss was observed in histological and biochemical analyses of different brain areas. The extent of prion replication was closely correlated with the up-regulation of ER-stress chaperone proteins. Similar results were observed in humans affected with sporadic and variant Creutzfeldt Jakob disease, implicating for the first time the caspase-12 dependent pathway in a neurodegenerative disease in vivo, and thus offering novel potential targets for the treatment of prion disorders. PMID- 14532117 TI - CHIP activates HSF1 and confers protection against apoptosis and cellular stress. AB - Induction of molecular chaperones is the characteristic protective response to environmental stress, and is regulated by a transcriptional program that depends on heat shock factor 1 (HSF1), which is normally under negative regulatory control by molecular chaperones Hsp70 and Hsp90. In metazoan species, the chaperone system also provides protection against apoptosis. We demonstrate that the dual function co-chaperone/ubiquitin ligase CHIP (C-terminus of Hsp70 interacting protein) regulates activation of the stress-chaperone response through induced trimerization and transcriptional activation of HSF1, and is required for protection against stress-induced apoptosis in murine fibroblasts. The consequences of this function are demonstrated by the phenotype of mice lacking CHIP, which develop normally but are temperature-sensitive and develop apoptosis in multiple organs after environmental challenge. CHIP exerts a central and unique role in tuning the response to stress at multiple levels by regulation of protein quality control and transcriptional activation of stress response signaling. PMID- 14532118 TI - Bcl-xL/Bcl-2 coordinately regulates apoptosis, cell cycle arrest and cell cycle entry. AB - Bcl-x(L) and Bcl-2 inhibit both apoptosis and proliferation. In investigating the relationship between these two functions of Bcl-x(L) and Bcl-2, an analysis of 24 Bcl-x(L) and Bcl-2 mutant alleles, including substitutions at residue Y28 previously reported to selectively abolish the cell cycle activity, showed that cell cycle delay and anti-apoptosis co-segregated in all cases. In determining whether Bcl-2 and Bcl-x(L) act in G(0) or G(1), forward scatter and pyronin Y fluorescence measurements indicated that Bcl-2 and Bcl-x(L) cells arrested more effectively in G(0) than controls, and were delayed in G(0)-G(1) transition. The cell cycle effects of Bcl-2 and Bcl-x(L) were reversed by Bad, a molecule that counters the survival function of Bcl-2 and Bcl-x(L). When control and Bcl-x(L) cells of equivalent size and pyronin Y fluorescence were compared, the kinetics of cell cycle entry were similar, demonstrating that the ability of Bcl-x(L) and Bcl-2 cells to enhance G(0) arrest contributes significantly to cell cycle delay. Our data suggest that cell cycle effects and increased survival both result from intrinsic functions of Bcl-2 and Bcl-x(L). PMID- 14532119 TI - Abl tyrosine kinases are required for infection by Shigella flexneri. AB - Infection by the opportunistic bacterial pathogen Shigella flexneri stimulates tyrosine phosphorylation of host cell proteins, but the kinases involved and their effects on the regulation of cell signaling pathways during bacterial entry remain largely undefined. Here, we demonstrate a requirement for the Abl family of tyrosine kinases during Shigella internalization. Family members Abl and Arg are catalytically activated upon Shigella infection, accumulate at the site of bacterial entry, and are required for efficient bacterial uptake, as internalization is blocked upon targeted deletion of these kinases or treatment with a specific pharmacological inhibitor. We identify the adapter protein Crk as a target for Abl kinases during Shigella uptake, and show that a phosphorylation deficient Crk mutant significantly inhibits bacterial uptake. Moreover, we define a novel signaling pathway activated during Shigella entry that links Abl kinase phosphorylation of Crk to activation of the Rho family GTPases Rac and Cdc42. Together, these findings reveal a new role for the Abl kinases, and suggest a novel approach to treatment of Shigella infections through inhibition of host cell signaling pathways. PMID- 14532120 TI - SCF(HOS) ubiquitin ligase mediates the ligand-induced down-regulation of the interferon-alpha receptor. AB - Down-regulation of activated signaling receptors in response to their ligands plays a key role in restricting the extent and duration of the signaling. Mechanisms underlying down-regulation of the type I interferon receptor consisting of IFNAR1 and IFNAR2 subunits remain largely unknown. Here we show that IFNAR1 interacts with the Homolog of Slimb (HOS) F-box protein in a phosphorylation-dependent manner, and that this interaction is promoted by interferon alpha (IFNalpha). IFNAR1 is ubiquitinated by the Skp1-Cullin1-HOS-Roc1 (SCF(HOS)) ubiquitin ligase in vitro. HOS expression and activities are required for IFNalpha-stimulated ubiquitination of IFNAR1, endocytosis of the type I interferon receptor, down-regulation of IFNAR1 levels, and IFNAR1 proteolysis via the lysosomal pathway. Furthermore, modulations of HOS activities affect the extent of Stat1 phosphorylation and Stat-mediated transcriptional activities as well as the extent of antiproliferative effects of type I interferons. These findings characterize SCF(HOS) as an E3 ubiquitin ligase that is essential for ubiquitination, proteolysis and down-regulation of IFNAR1, and implicate HOS in the regulation of cellular responses to IFNalpha. PMID- 14532121 TI - Activation of sphingosine kinase 1 by ERK1/2-mediated phosphorylation. AB - Sphingosine kinase 1 is an agonist-activated signalling enzyme that catalyses the formation of sphingosine 1-phosphate, a lipid second messenger that has been implicated in a number of agonist-driven cellular responses, including stimulation of cell proliferation, inhibition of apoptosis and expression of inflammatory molecules. Although agonist-induced stimulation of sphingosine kinase activity is critical in a number of signalling pathways, nothing has been known of the molecular mechanism of this activation. Here we show that this activation results directly from phosphorylation of sphingosine kinase 1 at Ser225, and present several lines of evidence to show compellingly that the activating kinase is ERK1/2 or a close relative. Furthermore, we show that phosphorylation of sphingosine kinase 1 at Ser225 results not only in an increase in enzyme activity, but is also necessary for translocation of the enzyme from the cytosol to the plasma membrane. Thus, these studies have elucidated the mechanism of agonist-mediated sphingosine kinase activation, and represent a key finding in understanding the regulation of sphingosine kinase/sphingosine 1 phosphate-controlled signalling pathways. PMID- 14532122 TI - Redox regulation of PI 3-kinase signalling via inactivation of PTEN. AB - The tumour suppressor PTEN is a PtdIns(3,4,5)P(3) phosphatase that regulates many cellular processes through direct antagonism of PI 3-kinase signalling. Here we show that oxidative stress activates PI 3-kinase-dependent signalling via the inactivation of PTEN. We use two assay systems to show that cellular PTEN phosphatase activity is inhibited by oxidative stress induced by 1 mM hydrogen peroxide. PTEN inactivation by oxidative stress also causes an increase in cellular PtdIns(3,4,5)P(3) levels and activation of the downstream PtdIns(3,4,5)P(3) target, PKB/Akt, that does not occur in cells lacking PTEN. We then show that endogenous oxidant production in RAW264.7 macrophages inactivates a fraction of the cellular PTEN, and that this is associated with an oxidant dependent activation of downstream signalling. These results show that oxidants, including those produced by cells, can activate downstream signalling via the inactivation of PTEN. This demonstrates a novel mechanism of regulation of the activity of this important tumour suppressor and the signalling pathways it regulates. These results may have significant implications for the many cellular processes in which PtdIns(3,4,5)P(3) and oxidants are produced concurrently. PMID- 14532123 TI - Determination of lymphoid cell fate is dependent on the expression status of the IL-7 receptor. AB - Signaling through the IL-7 receptor (IL-7R) is necessary for the development of the earliest B- and T-lineage cells. IL-7R is first expressed on common lymphoid progenitor cells and is not detected on primitive common myeloid progenitors. In this study, we show that enforced expression of IL-7R on multipotential stem cells does not influence lymphoid versus myeloid cell fate. T cell development was compatible with sustained IL-7R expression; however, we observed a near complete block in B cell development at the onset of B-lineage commitment. Unlike pre-proB cells from control animals, developmentally-arrested IL 7R(+)B220(+)CD19(-)NK1.1(-)Ly-6C(-) cells failed to express EBF and Pax5. These results suggest that transient downregulation of IL-7R signaling is a necessary event for induction of EBF and Pax5 expression and B-lymphocyte commitment. PMID- 14532124 TI - Groucho suppresses Pax2 transactivation by inhibition of JNK-mediated phosphorylation. AB - Pax proteins are DNA-binding transcription factors that regulate embryonic development through the activation and repression of downstream target genes. The Pax2 gene is absolutely required for kidney development and for patterning specific regions of the nervous system such as the eye, ear and hindbrain. The Pax2/5/8 family of proteins contains both transcription activation and repression domains. The activation domain of Pax2 is phosphorylated by the c-Jun N-terminal kinase (JNK) to enhance Pax2-dependent transcription. In this report, we demonstrate that the Groucho/TLE family protein, Grg4, interacts with Pax2 to suppress transactivation. Grg4 is able to specifically inhibit phosphorylation of the Pax2 activation domain, even in the presence of activated JNK. Furthermore, the Grg4 interaction and suppression of phosphorylation depends on Pax2 binding to its target DNA sequence and is independent of histone deacetylation. These data suggest a new model for Groucho mediated suppression of transcription through the specific inhibition of modifications in the activation domain of a transactivator. PMID- 14532125 TI - Genetic analysis of NF-kappaB/Rel transcription factors defines functional specificities. AB - The NF-kappaB transcription factors consist of dimeric proteins of the Rel homology family. They activate many promoters containing highly divergent kappaB site sequences. We have generated cell lines lacking individual and multiple NF kappaB proteins and used them to establish interactions between components of the NF-kappaB-IkappaB signaling system. Functional compensation within the family of dimers was evident in knockout cell lines. Analysis of transiently transfected genes gave an impression of promiscuity that was not borne out by analysis of endogenous genes. Using TNFalpha as an inducer, a panel of endogenous genes showed a wide range of subunit specificities as well as highly variable kinetics of induction. Comparing the function and subunit specificity of genes with the sequence of the kappaB DNA-binding site we found little correlation, indicating that NF-kappaB family member specificity for endogenous promoters is not solely encoded by the kappaB site sequence itself. PMID- 14532126 TI - Formation of facultative heterochromatin in the absence of HP1. AB - Facultative heterochromatin is a cytological manifestation of epigenetic mechanisms that regulate gene expression. Constitutive heterochromatin is marked by distinctive histone H3 methylation and the presence of HP1 proteins, but the chromatin modifications of facultative heterochromatin are less clear. We have examined histone modifications and HP1 in the facultative heterochromatin of nucleated erythrocytes and show that mouse and chicken erythrocytes have different mechanisms of heterochromatin formation. Mouse embryonic erythrocytes have abundant HP1, increased tri-methylation of H3 at K9 and loss of H3 tri methylation at K27. In contrast, we show that HP1 proteins are lost during the differentiation of chicken erythrocytes, and that H3 tri-methylation at both K9 and K27 is reduced. This coincides with the appearance of the variant linker histone H5. HP1s are also absent from erythrocytes of Xenopus and zebrafish. Our data show that in the same cell lineage there are different mechanisms for forming facultative heterochromatin in vertebrates. To our knowledge, this is the first report of cell types that lack HP1s and that have gross changes in the levels of histone modifications. PMID- 14532127 TI - Monocytic cells hyperacetylate chromatin protein HMGB1 to redirect it towards secretion. AB - High Mobility Group 1 protein (HMGB1) is a chromatin component that, when leaked out by necrotic cells, triggers inflammation. HMGB1 can also be secreted by activated monocytes and macrophages, and functions as a late mediator of inflammation. Secretion of a nuclear protein requires a tightly controlled relocation program. We show here that in all cells HMGB1 shuttles actively between the nucleus and cytoplasm. Monocytes and macrophages acetylate HMGB1 extensively upon activation with lipopolysaccharide; moreover, forced hyperacetylation of HMGB1 in resting macrophages causes its relocalization to the cytosol. Cytosolic HMGB1 is then concentrated by default into secretory lysosomes, and secreted when monocytic cells receive an appropriate second signal. PMID- 14532128 TI - Peripheral regions of natural hammerhead ribozymes greatly increase their self cleavage activity. AB - Natural hammerhead ribozymes are mostly found in some viroid and viroid-like RNAs and catalyze their cis cleavage during replication. Hammerheads have been manipulated to act in trans and assumed to have a similar catalytic behavior in this artificial context. However, we show here that two natural cis-acting hammerheads self-cleave much faster than trans-acting derivatives and other reported artificial hammerheads. Moreover, modifications of the peripheral loops 1 and 2 of one of these natural hammerheads induced a >100-fold reduction of the self-cleavage constant, whereas engineering a trans-acting artificial hammerhead into a cis derivative by introducing a loop 1 had no effect. These data show that regions external to the central conserved core of natural hammerheads play a role in catalysis, and suggest the existence of tertiary interactions between these peripheral regions. The interactions, determined by the sequence and size of loops 1 and 2 and most likely of helices I and II, must result from natural selection and should be studied in order to better understand the hammerhead requirements in vivo. PMID- 14532129 TI - A co-repressor assembly nucleated by Sex-lethal in the 3'UTR mediates translational control of Drosophila msl-2 mRNA. AB - Drosophila Sex-lethal (dSXL)-mediated translational repression of male-specific lethal 2 (msl-2) mRNA is essential for X-chromosome dosage compensation. Binding of dSXL to specific sites in both untranslated regions of msl-2 mRNA is necessary for inhibition of translation initiation. We describe the organization of dSXL as a translational regulator and show that the RNA binding and translational repressor functions are contained within the two RRM domains and a C-terminal heptapeptide extension. The repressor function is dormant unless dSXL binds to msl-2 mRNA with its own RRMs, because dSXL tethering via a heterologous RNA binding peptide does not elicit translational inhibition. We reveal proteins that crosslink to the msl-2 3' untranslated region (3'UTR) and co-immunoprecipitate with dSXL in a fashion that requires its intact repressor domain and correlates with translational regulation. Translation competition and UV-crosslink experiments show that the 3'UTR msl-2 sequences adjacent to dSXL-binding sites are necessary to recruit titratable co-repressors. Our data support a model where dSXL binding to the 3'UTR of msl-2 mRNA activates the translational repressor domain, thereby enabling it to recruit co-repressors in a specific fashion. PMID- 14532130 TI - Upstream AUGs in embryonic proinsulin mRNA control its low translation level. AB - Proinsulin is expressed prior to development of the pancreas and promotes cell survival. Here we study the mechanism affecting the translation efficiency of a specific embryonic proinsulin mRNA. This transcript shares the coding region with the pancreatic form, but presents a 32 nt extended leader region. Translation of proinsulin is markedly reduced by the presence of two upstream AUGs within the 5' extension of the embryonic mRNA. This attenuation is lost when the two upstream AUGs are mutated to AAG, leading to translational efficiency similar to that of the pancreatic mRNA. The upstream AUGs are recognized as initiator codons, because expression of upstream ORF is detectable from the embryonic transcript, but not from the mutated or the pancreatic mRNAs. Strict regulation of proinsulin biosynthesis appears to be necessary, since exogenous proinsulin added to embryos in ovo decreased apoptosis and generated abnormal developmental traits. A novel mechanism for low level proinsulin expression thus relies on upstream AUGs within a specific form of embryonic proinsulin mRNA, emphasizing its importance as a tightly regulated developmental signal. PMID- 14532131 TI - The roles of initiation factor 2 and guanosine triphosphate in initiation of protein synthesis. AB - The role of IF2 from Escherichia coli was studied in vitro using a system for protein synthesis with purified components. Stopped flow experiments with light scattering show that IF2 in complex with guanosine triphosphate (GTP) or a non cleavable GTP analogue (GDPNP), but not with guanosine diphosphate (GDP), promotes fast association of ribosomal subunits during initiation. Biochemical experiments show that IF2 promotes fast formation of the first peptide bond in the presence of GTP, but not GDPNP or GDP, and that IF2-GDPNP binds strongly to post-initiation ribosomes. We conclude that the GTP form of IF2 accelerates formation of the 70S ribosome from subunits and that GTP hydrolysis accelerates release of IF2 from the 70S ribosome. The results of a recent report, suggesting that GTP and GDP promote initiation equally fast, have been addressed. Our data, indicating that eIF5B and IF2 have similar functions, are used to rationalize the phenotypes of GTPase-deficient mutants of eIF5B and IF2. PMID- 14532132 TI - A replication silencer element in a plus-strand RNA virus. AB - Replication represents a key step in the infectious cycles of RNA viruses. Here we describe a regulatory RNA element, termed replication silencer, that can down regulate complementary RNA synthesis of a positive-strand RNA virus via an RNA RNA interaction. This interaction occurs between the 5-nucleotide-long, internally positioned replication silencer and the extreme 3'-terminus of the viral RNA comprising part of the minimal minus-strand initiation promoter. Analysis of RNA synthesis in vitro, using model defective interfering (DI) RNA templates of tomato bushy stunt virus and a partially purified, RNA-dependent RNA polymerase preparation from tombusvirus-infected plants, revealed that this interaction inhibits minus-strand synthesis 7-fold. This functional interaction was supported further by: (i) RNA structure probing; (ii) phylogenetic analysis; (iii) inhibition of activity by short complementary DNAs; and (iv) compensatory mutational analysis. The silencer was found to be essential for accumulation of DI RNAs in protoplasts, indicating that it serves an important regulatory role(s) in vivo. Because similar silencer-promoter interactions are also predicted in other virus genera, this type of RNA-based regulatory mechanism may represent a widely utilized strategy for modulating replication. PMID- 14532134 TI - Dmp53 protects the Drosophila retina during a developmentally regulated DNA damage response. AB - Ultraviolet (UV) light is absorbed by cellular proteins and DNA, promoting skin damage, aging and cancer. In this paper, we explore the UV response by cells of the Drosophila retina. We demonstrate that the retina enters a period of heightened UV sensitivity in the young developing pupa, a stage closely associated with its period of normal developmental programmed cell death. Injury to irradiated cells included morphology changes and apoptotic cell death; these defects could be completely accounted for by DNA damage. Cell death, but not morphological changes, was blocked by the caspase inhibitor P35. Utilizing genetic and microarray data, we provide evidence for the central role of Hid expression and for Diap1 protein stability in controlling the UV response. In contrast, we found that Reaper had no effect on UV sensitivity. Surprisingly, Dmp53 is required to protect cells from UV-mediated cell death, an effect attributed to its role in DNA repair. These in vivo results demonstrate that the cellular effects of DNA damage depend on the developmental status of the tissue. PMID- 14532133 TI - Requirement of the MRN complex for ATM activation by DNA damage. AB - The ATM protein kinase is a primary activator of the cellular response to DNA double-strand breaks (DSBs). In response to DSBs, ATM is activated and phosphorylates key players in various branches of the DNA damage response network. ATM deficiency causes the genetic disorder ataxia-telangiectasia (A-T), characterized by cerebellar degeneration, immunodeficiency, radiation sensitivity, chromosomal instability and cancer predisposition. The MRN complex, whose core contains the Mre11, Rad50 and Nbs1 proteins, is involved in the initial processing of DSBs. Hypomorphic mutations in the NBS1 and MRE11 genes lead to two other genomic instability disorders: the Nijmegen breakage syndrome (NBS) and A-T like disease (A-TLD), respectively. The order in which ATM and MRN act in the early phase of the DSB response is unclear. Here we show that functional MRN is required for ATM activation, and consequently for timely activation of ATM-mediated pathways. Collectively, these and previous results assign to components of the MRN complex roles upstream and downstream of ATM in the DNA damage response pathway and explain the clinical resemblance between A-T and A-TLD. PMID- 14532135 TI - Distinct regulators for Plk1 activation in starfish meiotic and early embryonic cycles. AB - The Polo-like kinase, Plk, has multiple roles in regulating mitosis. In particular, Plk1 has been postulated to function as a trigger kinase that phosphorylates and activates Cdc25C prior to the activation of cyclin B-Cdc2 and thereby initiates its activation. However, the upstream regulation of Plk1 activation remains unclear. Here we have studied the interplay between Plk1 and Cdc2 through meiotic and early embryonic cycles in starfish. Distinct kinases, cyclin B-Cdc2, MAPK along with cyclin B- and/or cyclin A-Cdc2 and cyclin A-Cdc2, were unique upstream regulators for Plk1 activation at meiosis I, meiosis II and embryonic M-phase, respectively, indicating that Plk1 is not the trigger kinase at meiotic reinitiation. When Plk1 was required for cyclin B-Cdc2 activation, the action of Plk1 was mediated primarily through suppression of Myt1 rather than through activation of Cdc25. We propose that Plk1 can be activated by either cyclin A- or cyclin B-Cdc2, and its primary target is Myt1. PMID- 14532137 TI - Advances in targeted therapies V: Proceedings of the 5th International Symposium. St Martin, French West Indies. 11-14 April 2003. PMID- 14532136 TI - Rec8 cleavage by separase is required for meiotic nuclear divisions in fission yeast. AB - Sister chromatid cohesion in meiosis is established by cohesin complexes, including the Rec8 subunit. During meiosis I, sister chromatid cohesion is destroyed along the chromosome arms to release connections of recombined homologous chromosomes (homologues), whereas centromeric cohesion persists until it is finally destroyed at anaphase II. In fission yeast, as in mammals, distinct cohesin complexes are used depending on the chromosomal region; Rec8 forms a complex with Rec11 (equivalent to SA3) mainly along chromosome arms, while Psc3 (equivalent to SA1 and SA2) forms a complex mainly in the vicinity of the centromeres. Here we show that separase activation and resultant Rec8 cleavage are required for meiotic chromosome segregation in fission yeast. A non-cleavable form of Rec8 blocks disjunction of homologues at meiosis I. However, displacing non-cleavable Rec8 restrictively from the chromosome arm by genetically depleting Rec11 alleviated the blockage of homologue segregation, but not of sister segregation. We propose that the segregation of homologues at meiosis I and of sisters at meiosis II requires the cleavage of Rec8 along chromosome arms and at the centromeres, respectively. PMID- 14532139 TI - Imaging: do erosions heal? AB - It was shown previously that reparative changes in erosions can be seen in individual joints and individual patients. Whether repair may occur at a group level, and can be induced by treatment, is not known. This manuscript describes a means of visualising data obtained in a clinical trial by the use of probability plots to better understand the results. These probability plots give a good insight into the coherence of the data. They can also be used to make the interpretation of repair at a group level easier. Probability plots also explain the hazard of using binomial cut off points to compare treatment effects. The interference of true repair with measurement error is demonstrated. Repair at a group level is suggested if the mean progression score is statistically significantly different from zero, which can be visualised by a 95% confidence interval of this mean change score below zero. Application of this technique may give us better information on the effects of new drugs on the induction of repair. PMID- 14532138 TI - Updated consensus statement on biological agents for the treatment of rheumatoid arthritis and other immune mediated inflammatory diseases (May 2003). PMID- 14532140 TI - Comparison of the efficacy of the tumour necrosis factor alpha blocking agents adalimumab, etanercept, and infliximab when added to methotrexate in patients with active rheumatoid arthritis. AB - OBJECTIVE: To determine, using the method of adjusted indirect comparisons, whether there are differences in efficacy of tumour necrosis factor (TNF alpha) blocking agents, as measured by the rate ratios for American College of Rheumatology (ACR) 20, 50, and 70 responses, in patients with rheumatoid arthritis with an incomplete response to methotrexate. METHODS: A systematic review was performed to identify placebo controlled trials of 24-30 weeks' duration of combination therapy that used a step-up approach with the addition of TNF alpha blocking agents to methotrexate. The method of "adjusted indirect comparisons" was used to compare results across trials to determine the relative risk for an ACR20 or 50 response. RESULTS: Placebo controlled trials for adalimumab, etanercept, and infliximab provided data on ACR20 and 50 responses. Both the similarity of demographic and disease characteristics of patients at entry, and the homogeneity of response rates in the placebo groups across trials, suggested that comparing results across trials was valid. The relative risk for obtaining an ACR20 and 50 response derived from the adjusted indirect comparisons of the TNF alpha blocking agents did not significantly differ from unity. CONCLUSION: The analysis showed that the three currently marketed TNF alpha blocking agents have similarly efficacy when added to methotrexate in the treatment of patients with rheumatoid arthritis with active disease. PMID- 14532141 TI - Roles of antimicrobial peptides such as defensins in innate and adaptive immunity. AB - A number of antimicrobial peptides such as defensins have multiple functions in host defence. Defensins are produced not only by phagocytic cells and lymphocytes, but also by the epithelial cell lining of the gastrointestinal and genitourinary tracts, the tracheobronchial tree, and keratinocytes. Some are produced constitutively, whereas others are induced by proinflammatory cytokines and exogenous microbial products. Defensins produced by cells in the course of innate host defence serve as signals which initiate, mobilise, and amplify adaptive immune host defences. Administration of defensins with antigens to mice enhances both cellular (Th1-dependent) and humoral (Th2-dependent) cytokine production and immune responses. Linkage of defensins to weak tumour antigens potentiates their immunoadjuvant effects. Defensins use multiple cellular receptors, which endows them with the capacity to marshall adaptive host defences against microbial invaders. PMID- 14532142 TI - Possible targeting of G protein coupled receptors to manipulate inflammation in vivo using synthetic and natural ligands. AB - Cyclic AMP elevating Gs protein coupled receptors were considered for a long time to be immunosuppressive. One of these receptors, adenosine A(2A) receptor, was implicated in a physiological mechanism that down regulates inflammation and protects tissues from excessive immune mediated damage. Targeting of these receptors by selective agonists may lead to better protocols of anti-inflammatory treatments. At the same time inhibiting the Gs protein coupled mediated signalling with antagonists could be explored in studies of approaches to enhance inflammation and tissue damage. Enhancement of targeted tissue damage is highly desirable when it is cancerous tissue, while enhancement of inflammatory events might be desirable in the development of new vaccine adjuvants. PMID- 14532143 TI - Actions of BAFF in B cell maturation and its effects on the development of autoimmune disease. AB - BAFF, a member of the family of tumour necrosis factor (TNF) ligands, is essential for the development of peripheral mature, long lived B lymphocytes. It binds to three different receptors, BCMA, TACI, and BAFF-R, which are all members of the family of TNF receptors. Defects in the genes encoding BAFF or BAFF-R abolish the generation of mature B cells. BAFF is made by myeloid cells whereas BAFF-R is expressed preferentially on B cells. BAFF induces polyclonal maturation of resting, short lived immature B cells to resting, long lived mature B cells without proliferation. Lupus erythematodes prone mice have elevated blood levels of BAFF, and treatment of these mice with the BAFF decoy receptor (BCMA-Ig) prevents the onset of this systemic autoimmune disease. Human lupus patients also have elevated blood levels of BAFF. Treatment with BAFF neutralising agents (decoy receptors, monoclonal antibodies) should prevent, delay, or, at least, slow down the disease. PMID- 14532144 TI - British Society for Rheumatology Biologics Register. AB - The British Society for Rheumatology (BSR) established a register of patients newly treated with biological agents, the BSR Biologics Register (BSRBR), which became active in January 2002. The goal is to register all patients in the United Kingdom with rheumatic diseases, newly starting treatment with these agents and to follow them up to determine the incidence of any short and long term hazards to health. The Register is also recruiting a comparison cohort of patients with rheumatoid arthritis treated with standard disease modifying antirheumatic drugs to determine the relative contributions of disease factors and other treatments apart from biological agents on any risks observed. PMID- 14532145 TI - Survival during treatment with tumour necrosis factor blocking agents in rheumatoid arthritis. AB - Tumour necrosis factor (TNF) blocking agents are an important advance in the clinical treatment of rheumatoid arthritis (RA). They were introduced into clinical practice while limited safety information was available. This means that intensive monitoring is needed early in the life cycle of these new drugs. Setting up large cohort studies to monitor efficacy, safety, and tolerability in long term use of these so-called biological agents will provide information about the consequences of using TNF blocking agents in chronic rheumatic disease like RA. Currently, a Dutch multicentre registry on biological agents in RA is being set up. This study aimed at investigating the efficacy and toxicity of TNF blocking agents in patients with RA at one participating academic centre by a drug survival analysis. Since 1997 230 patients with RA at the centre have been treated with TNF blocking agents for the first time (94 with adalimumab, 120 with infliximab, and 16 with etanercept). No differences in drug survival between the three TNF blocking agents were found despite the diversity in selection and patient numbers. Adverse events which occurred, leading to discontinuation, were similar to those from previous reports. PMID- 14532146 TI - Advances in targeted therapy: safety of biological agents. PMID- 14532147 TI - Role of tumour necrosis factor (TNF) in host defence against tuberculosis: implications for immunotherapies targeting TNF. AB - Studies in mouse infection models clearly demonstrate tumour necrosis factor (TNF) to be a critical component of both the antibacterially protective and the inflammatory immune response to Mycobacterium tuberculosis. It is therefore not surprising that treatment of patients-for example, those with rheumatoid arthritis-with biological agents interfering with TNF activity have shown an increased risk of reactivating tuberculosis. However, conceivably, TNF targeting biological agents can be developed that because of their particular mode of action and their specific pharmacodynamics may be less likely to have this side effect. PMID- 14532148 TI - Metalloproteinases, inflammation, and rheumatoid arthritis. AB - Ideally, the inflammatory response occurs rapidly to terminate infection. It also must halt in a timely manner to stop this reaction from inflicting self damage. Such a highly regulated process results from altering balances in pro- and anti inflammatory signals orchestrated by multiple cell types and factors within the tissue microenvironment. The discovery of new substrates of metalloproteinases within this microenvironment has disclosed a new function in inflammation. The role of these proteases now extends beyond extracellular matrix remodelling enzymes to that of mediators of inflammatory signals involving various chemokines and cytokines. As natural inhibitors of these metalloproteinases, TIMPs have the potential of regulating the inflammatory response and affecting diseases such as rheumatoid arthritis. TIMP-3, in particular, stands out as an important regulator of inflammation with its ability to specifically inhibit proinflammatory cytokines and tissue destruction in the joint. PMID- 14532149 TI - Role of interleukin 18 in rheumatoid arthritis. PMID- 14532150 TI - New strategies to control inflammatory synovitis: interleukin 15 and beyond. PMID- 14532151 TI - B cell therapy for rheumatoid arthritis: the rituximab (anti-CD20) experience. PMID- 14532153 TI - Interleukin 6 receptor as a target for the treatment of rheumatoid arthritis. PMID- 14532154 TI - Alpha 4 integrin blockade in inflammatory bowel disease. PMID- 14532152 TI - Angiogenesis as a target in rheumatoid arthritis. PMID- 14532155 TI - Cartilage morphogenetic proteins: role in joint development, homoeostasis, and regeneration. AB - BACKGROUND: Articular cartilage homoeostasis is critical for joint function. The steady state homoeostasis of articular cartilage is a balance between anabolic morphogens such as cartilage derived morphogenetic proteins (CDMPs) and bone morphogenetic proteins (BMPs) of the BMP family and catabolic cytokines such as interleukin (IL)1, IL17, and tumour necrosis factor alpha. Although bone and articular cartilage are adjacent tissues, there is a profound difference in their regeneration potential. Bone has the highest potential for regeneration. On the other hand, articular cartilage is recalcitrant to repair. OBJECTIVE: To examine the hypothesis that the feeble innate regeneration ability of cartilage is due to the preponderance of catabolic cytokines such as IL1 and IL17. RESULTS: During a systematic investigation of CDMPs and cytokines IL17B (chondroleukin) was found in bovine articular cartilage. DISCUSSION AND CONCLUSIONS: BMP-7 and IL17B are present in articular cartilage and synthesised in chondrocytes as shown by northern blots and real-time reverse transcription-polymerase chain reaction. The coexistence of anabolic morphogens and catabolic cytokines in articular cartilage has important implications for cartilage homoeostasis and regeneration. The networks of signalling systems of morphogens and cytokines determine the net capacity for regenerative morphogenesis of articular cartilage. Finally, the feeble innate capacity for articular cartilage may be improved by targeted therapy by soluble receptors to block catabolic cytokines. PMID- 14532156 TI - Therapeutic targets in osteoarthritis: from today to tomorrow with new imaging technology. PMID- 14532157 TI - Signalling in osteoclasts and the role of Fos/AP1 proteins. PMID- 14532158 TI - Signal transduction networks in rheumatoid arthritis. AB - Signal transduction pathways regulate cellular responses to stress and play a critical role in inflammation. The complexity and specificity of signalling mechanisms represent major hurdles for developing effective, safe therapeutic interventions that target specific molecules. One approach is to dissect the pathways methodically to determine their hierarchy in various cell types and diseases. This approach contributed to the identification and prioritisation of specific kinases that regulate NF-kappa B and the mitogen activated protein (MAP) kinase cascade as especially attractive targets. Although significant issues remain with regard to the discovery of truly selective kinase inhibitors, the risks that accompany inhibition of fundamental signal transduction mechanisms can potentially be decreased by careful dissection of the pathways and rational target selection. PMID- 14532159 TI - Prospects for the development of small molecular weight compounds to replace anti tumour necrosis factor biological agents. PMID- 14532160 TI - Rheumatoid arthritis in 2003: where are we now with treatment? PMID- 14532161 TI - Bone buffering of acid and base in humans. AB - The sources and rates of metabolic acid production in relation to renal net acid excretion and thus acid balance in humans have remained controversial. The techniques and possible errors in these measurements are reviewed, as is the relationship of charge balance to acid balance. The results demonstrate that when acid production is experimentally increased among healthy subjects, renal net acid excretion does not increase as much as acid production so that acid balances become positive. These positive imbalances are accompanied by equivalently negative charge balances that are the result of bone buffering of retained H+ and loss of bone Ca2+ into the urine. The data also demonstrate that when acid production is experimentally reduced during the administration of KHCO3, renal net acid excretion does not decrease as much as the decrease in acid production so that acid balances become negative, or, in opposite terms, there are equivalently positive HCO3- balances. Equivalently positive K+ and Ca2+ balances, and thus positive charge balances, accompany these negative acid imbalances. Similarly, positive Na+ balances, and thus positive charge balances, accompany these negative acid balances during the administration of NaHCO3. These charge balances are likely the result of the adsorption of HCO3- onto the crystal surfaces of bone mineral. There do not appear to be significant errors in the measurements. PMID- 14532162 TI - Noncoordinate regulation of ENaC: paradigm lost? AB - The epithelial sodium channel (ENaC) is composed of the three homologous subunits alpha, beta, and gamma. The basic oligomerization process inferred from all studies in heterologous systems is preferential assembly of the three subunits into a single oligomeric form. However, there is also considerable evidence that channels composed of only alpha-, alphabeta-, or alphagamma-subunits can form under some circumstances and that individual subunits expressed in heterologous systems can traffic to the cell membrane. In cells that express endogenous ENaC, the three subunits are often synthesized in a differential fashion, with one or two subunits expressed constitutively while the other(s) are induced by different physiological stimuli in parallel with increased ENaC activity. This phenomenon, which we term noncoordinate regulation, has been observed for both whole cell and apical membrane ENaC subunit expression. Several other heteromeric membrane proteins have also been observed to have differential rates of either turnover or trafficking of individual subunits after biosynthesis and membrane localization. Here, we examine the possibility that noncoordinate regulation of ENaC subunits may represent another mechanism in the arsenal of physiological responses to diverse stimuli. PMID- 14532163 TI - Activity of the basolateral K+ channels is coupled to the Na+-K+-ATPase in the cortical collecting duct. AB - Microelectrode and patch-clamp techniques were used in the isolated cortical collecting duct to study the effects of stimulating Na+-K+-ATPase by raising bath K+ (Fujii Y and Katz AI. Am J Physiol Renal Fluid Electrolyte Physiol 257: F595 F601, 1989 and Muto S, Asano Y, Seldin D, and Giebisch. Am J Physiol Renal Physiol 276: F143-F158, 1999) on the transepithelial (VT) and basolateral membrane (VB) voltages and basolateral K+ channel activity. Increasing bath K+ from 2.5 to 8.5 mM resulted in an initial hyperpolarization of both VT and VB followed by a delayed depolarization. The effects of raising bath K+ on VT and VB were attenuated by decreasing luminal Na+ from 146.8 to 14.0 mM and were abolished by removal of luminal Na+, whereas those were magnified in desoxycorticosterone acetate (DOCA)-treated rabbits. Increasing bath K+ also led to a significant reduction of the intracellular Na+ and Ca2+ concentrations. The transepithelial conductance (GT) or fractional apical membrane resistance (fRA) were unaltered during the initial hyperpolarization phase, whereas, in the late depolarization phase, there were an increase in GT and a decrease in fRA, both of which were attenuated in the presence of low luminal Na+ (14.0 mM). In tubules from DOCA-treated animals, bath Ba2+ not only caused a significantly larger initial hyperpolarization of VT and VB but also blunted the late depolarization by high bath K+. Nomega-nitro-l-arginine methyl ester (l-NAME) partially mimicked the effect of Ba2+ and decreased the amplitude of the late depolarization. Patch clamp experiments showed that raising bath K+ from 2.5 to 8.5 mM resulted in an increased activity of the basolateral K+ channel, which was absent in the presence of l-NAME. We conclude that stimulation of Na+-K+-ATPase increases the basolateral K+ conductance and that this effect involves suppression of nitric oxide-dependent inhibition of K+ channels. PMID- 14532164 TI - Proteomic analysis of long-term vasopressin action in the inner medullary collecting duct of the Brattleboro rat. AB - Vasopressin regulates water and solute transport in the renal collecting duct. In addition to short-term regulation of aquaporin-2 trafficking, vasopressin also has long-term effects to regulate the abundances of aquaporins-2 and -3 and beta- and gamma-subunits of the epithelial sodium channel in collecting duct principal cells. To investigate further the direct and indirect long-term regulatory actions of vasopressin in the inner medullary collecting duct (IMCD), we used a proteomic approach [difference gel electrophoresis (DIGE) coupled with MALDI-TOF identification of differentially expressed protein spots]. DDAVP or vehicle was infused subcutaneously in Brattleboro rats for 3 days, and IMCD cells were purified from the inner medullas for proteomic analysis. Forty-three proteins were found to be regulated in response to vasopressin infusion, including 18 that were increased in abundance, 22 that were decreased, and 3 that were shifted in the gel, presumably because of posttranslational modification. Immunocytochemistry confirmed collecting duct expression of several of the proteins that were identified. Immunoblot analysis of nine of the proteins confirmed the changes seen by the DIGE method. Of these nine proteins, six were increased in response to DDAVP infusion: nitric oxide synthase-2 (NOS2), GRP78, heat shock protein-70, annexin II, glutaminase, and cathepsin D. The remaining three were decreased in response to DDAVP: aldehyde reductase I, adenylyl cyclase VI, and carbonic anhydrase II. The findings point to a role for vasopressin in the coordinate regulation of several determinants of nitric oxide levels (NOS2, arginase II, NADPH oxidase) and of proteins potentially involved in vasopressin escape (adenylyl cyclase VI and G protein-coupled receptor kinase 4). PMID- 14532165 TI - Increased systemic and adipose tissue cytokines in patients with HIV-associated lipodystrophy. AB - The lipodystrophy syndrome (adipose tissue redistribution and metabolic abnormalities) observed with highly active antiretroviral therapy (HAART) during human immunodeficiency virus (HIV) infection may be related to increased proinflammatory cytokine activity. We measured acute cytokine (TNF-alpha, IL-6, leptin), glycerol, and lactate secretion from abdominal subcutaneous adipose tissue (SAT), and systemic cytokine levels, in HIV-infected subjects with and without lipodystrophy (HIVL+ and HIVL-, respectively) and healthy non-HIV controls. Lipodystrophy was confirmed and characterized as adipose tissue redistribution in HIVL+ compared with HIVL- and controls, by dual-energy X-ray absorptiometry and by whole body MRI. TNF-alpha secretion from abdominal SAT and circulating levels of IL-6, soluble TNF receptors I and II, and insulin were elevated in HIVL+ relative to HIVL- and/or controls, particularly in HIVL+ undergoing HAART. In the HIV-infected group as a whole, IL-6 secretion from abdominal SAT and serum IL-6 were positively associated with visceral fat and were negatively associated with the relative amount of lower limb adipose tissue (P < 0.01). Decreased leptin and increased lactate secretion from abdominal SAT were specifically associated with HAART. In conclusion, increased cytokine secretion from adipose tissue and increased systemic proinflammatory cytokine activity may play a significant role in the adipose tissue remodeling and/or the metabolic abnormalities associated with the HIV-lipodystrophy syndrome in patients undergoing HAART. PMID- 14532166 TI - Comparison of intraduodenal and intravenous glucose metabolism under clamp conditions in humans. AB - To determine whether the uptake and metabolic partition of glucose are influenced by its delivery route, 12 normal volunteers underwent two 3-h euglycemic (approximately 93 mg/dl) hyperinsulinemic (approximately 43 mU/l) clamps at a 3- to 5-wk interval, one with intravenous (i.v.) and the other with intraduodenal (i.d.) glucose labeled with [3-3H]- and [U-14C]glucose. Systemic glucose was traced with [6,6-2H2]glucose in eight subjects. During the last hour of the clamps, the average glucose infusion rate (5.85 +/- 0.37 vs. 5.43 +/- 0.43 mg.kg( 1).min(-1); P = 0.02) and exogenous glucose uptake (5.66 +/- 0.37 vs. 5.26 +/- 0.41 mg.kg(-1).min(-1); P = 0.04) were borderline higher in the i.d. than in the i.v. studies. The increased uptake was entirely accounted for by increased glycolysis (3H2O production), which was attributed to the stimulation of gut metabolism by the absorptive process. No difference was observed in glucose storage whether it was calculated as glucose uptake minus glycolysis (i.d. vs. i.v.: 2.44 +/- 0.28 vs. 2.40 +/- 0.31 mg.kg(-1).min(-1)) or as glucose uptake minus net glucose oxidation (2.86 +/- 0.33 vs. 2.81 +/- 0.35 mg.kg(-1).min(-1)). Because peripheral tissues were exposed to identical glucose, insulin, and free fatty acid levels under the two experimental conditions, we assumed that their glucose uptake and storage were similar during the two tests. We therefore suggest that hepatic glycogen storage (estimated as whole body minus peripheral storage) was also unaffected by the route of glucose delivery. On the other hand, in the i.d. tests, the glucose splanchnic extraction ratio calculated by the dual isotope technique averaged 4.9 +/- 2.3%, which is close to the figures published for i.v. glucose. Despite the limitations related to whole body measurements, these two sets of data do not support the idea that enteral glucose stimulates hepatic uptake more efficiently than i.v. glucose. PMID- 14532167 TI - Body cell mass: model development and validation at the cellular level of body composition. AB - Existing models to estimate the metabolically active body cell mass (BCM) component in vivo remain incompletely developed. The classic Moore model is based on an assumed BCM potassium content of 120 mmol/kg. Our objectives were to develop an improved total body potassium (TBK)-independent BCM prediction model on the basis of an earlier model (Cohn SH, Vaswani AN, Yasumura S, Yuen K, and Ellis KJ. J Lab Clin Med 105: 305-311, 1985), to apply this improved model in subjects to explore the sex and age dependence of the TBK/BCM ratio, to develop a new TBK/BCM model on the basis of physiological associations between TBK and total body water (TBW) at the cellular level of body composition, and to fit this new model with available reference data. Subjects were 112 healthy adults who had the following components measured: TBW by 2H2O or 3H2O, extracellular water by NaBr, total body nitrogen by in vivo neutron activation, bone mineral by dual energy X-ray absorptiometry, and TBK by whole body counting. Human reference data were collected from earlier published reports. The improved Cohn model-derived TBK/BCM ratio was (mean +/- SD) 109.0 +/- 10.9 mmol/kg and was not significantly related to sex and age. A simplified version of the new TBK-TBW model provided a TBK/BCM ratio almost identical (109.1 mmol/kg) to that derived by the improved Cohn model. The TBK-BCM prediction formula derived from the improved and new models [BCM (kg) = 1/109 x TBK (mmol); or BCM = 0.0092 x TBK] gives BCM estimates approximately 11% higher than the classic Moore model (BCM = 0.0083 x TBK) formulated on rough tissue composition estimates. The present analyses provide a physiologically based, improved, and validated TBK-BCM prediction formula that should prove useful in body composition and metabolism research. PMID- 14532168 TI - Insulin stimulates interleukin-6 and tumor necrosis factor-alpha gene expression in human subcutaneous adipose tissue. AB - High circulating levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) are found in patients with hyperinsulinemia. Insulin stimulates release of IL-6 from adipocyte cultures, and it stimulates IL-6 gene expression in insulin-resistant, but not control, rat skeletal muscle. In addition, TNF alpha may be involved in the pathogenesis of insulin resistance. Therefore, we studied the effect of insulin on IL-6 and TNF-alpha gene expression in human skeletal muscle and adipose tissue. Nine healthy young volunteers participated in the study. They underwent a 6-h hyperinsulinemic euglycemic clamp at a fixed insulin infusion rate, with blood glucose clamped at fasting level. Blood samples drawn at 0, 1, 2, 3, 4, 5, and 6 h were analyzed for IL-6 and TNF-alpha. Muscle and fat biopsies, obtained at 0, 2, 4, and 6 h, were analyzed for IL-6 and TNF alpha mRNA with real-time PCR. IL-6 mRNA increased 11-, 3-, and 5-fold at 2, 4, and 6 h, respectively, in adipose tissue (ANOVA P = 0.027), whereas there was no significant effect of insulin on skeletal muscles. Plasma IL-6 increased during insulin stimulation. TNF-alpha mRNA increased 2.4-, 1.4-, and 2.2-fold in adipose tissue (ANOVA P = 0.001) and decreased 0.74-, 0.64-, and 0.68-fold in muscle tissue (ANOVA P = 0.04). Plasma levels of TNF-alpha were constant. In conclusion, the finding that insulin stimulates IL-6 and TNF-alpha gene expression in adipose tissue only and inhibits the TNF-alpha production in skeletal muscles suggests a differential regulation of muscle- and adipose tissue-derived IL-6 and TNF-alpha. PMID- 14532169 TI - Impact of infection on glucose-dependent liver glucose uptake during TPN: interaction with insulin. AB - Chronic total parenteral nutrition (TPN) markedly augments net hepatic glucose uptake (NHGU). This adaptive increase is impaired by an infection despite accompanying hyperinsulinemia. In the nonadapted state, NHGU is dependent on the prevailing glucose levels. Our aims were to determine whether the adaptation to TPN alters the glucose dependence of NHGU, whether infection impairs this dependence, and whether insulin modulates the glucose dependence of NHGU during infection. Chronically catheterized dogs received TPN for 5 days. On day 3 of TPN, dogs received either a bacterial fibrin clot to induce a nonlethal infection (INF, n = 9) or a sterile fibrin clot (Sham, n = 6). Forty-two hours after clot implantation, somatostatin was infused. In Sham, insulin and glucagon were infused to match the level seen in Sham (9 +/- 1 microU/ml and 23 +/- 4 pg/ml, respectively). In infected animals, either insulin and glucagon were infused to match the levels seen in infection (25 +/- 2 microU/ml and 101 +/- 15 pg/ml; INF HI; n = 5) or insulin was replaced to match the lower levels seen in Sham (13 +/- 2 microU/ml), whereas glucagon was kept elevated (97 +/- 9 pg/ml; INF-LO; n = 4). Then a four-step (90 min each) hyperglycemic (120, 150, 200, or 250 mg/dl) clamp was performed. NHGU increased at each glucose step in Sham (from 3.6 +/- 0.6 to 5.4 +/- 0.7 to 8.9 +/- 0.9 to 12.1 +/- 1.1 mg.kg(-1).min(-1)); the slope of the relationship between glucose levels and NHGU (i.e., glucose dependence) was higher than that seen in nonadapted animals. Infection impaired glucose-dependent NHGU in both INF-HI (1.3 +/- 0.4 to 2.9 +/- 0.5 to 5.5 +/- 1.0 to 7.7 +/- 1.6 mg.kg(-1).min(-1)) and INF-LO (0.5 +/- 0.7 to 2.2 +/- 0.6 to 4.2 +/- 1.0 to 5.8 +/- 0.8 mg.kg(-1).min(-1)). In summary, TPN augments glucose-dependent NHGU, the presence of infection decreases glucose-dependent NHGU, and the accompanying hyperinsulinemia associated with infection does not sustain the glucose dependence of NHGU. PMID- 14532170 TI - AMPK activity and isoform protein expression are similar in muscle of obese subjects with and without type 2 diabetes. AB - Acute or chronic activation of AMP-activated protein kinase (AMPK) increases insulin sensitivity. Conversely, reduced expression and/or function of AMPK might play a role in insulin resistance in type 2 diabetes. Thus protein expression of the seven subunit isoforms of AMPK and activities and/or phosphorylation of AMPK and acetyl-CoA carboxylase-beta (ACCbeta) was measured in skeletal muscle from obese type 2 diabetic and well-matched control subjects during euglycemic hyperinsulinemic clamps. Protein expression of all AMPK subunit isoforms (alpha1, alpha2, beta1, beta2, gamma1, gamma2, and gamma3) in muscle of obese type 2 diabetic subjects was similar to that of control subjects. In addition, alpha1- and alpha2-associated activities of AMPK, phosphorylation of alpha-AMPK subunits at Thr172, and phosphorylation of ACCbeta at Ser221 showed no difference between the two groups and were not regulated by physiological concentrations of insulin. These data suggest that impaired insulin action on glycogen synthesis and lipid oxidation in skeletal muscle of obese type 2 diabetic subjects is unlikely to involve changes in AMPK expression and activity. PMID- 14532171 TI - Pioglitazone preserves pancreatic islet structure and insulin secretory function in three murine models of type 2 diabetes. AB - Thiazolidinediones may slow the progression of type 2 diabetes by preserving pancreatic beta-cells. The effects of pioglitazone (PIO) on structure and function of beta-cells in KKA(y), C57BL/6J ob/ob, and C57BL/KsJ db/db mice (genetic models of type 2 diabetes) were examined. ob/ob (n = 7) and db/db (n = 9) mice were randomly assigned to 50-125 mg.kg body wt-1.day-1 of PIO in chow beginning at 6-10 wk of age. Control ob/ob (n = 7) and db/db mice (n = 9) were fed chow without PIO. KKA(y) mice (n = 15) were fed PIO daily at doses of 62-144 mg.kg body wt-1.day-1. Control KKA(y) mice (n = 10) received chow without PIO. Treatment continued until euthanasia at 14-26 wk of age. Blood was collected at baseline (before treatment) and just before euthanasia and was analyzed for glucose, glycosylated hemoglobin, and plasma insulin. Some of the splenic pancreas of each animal was resected and partially sectioned for light or electron microscopy. The remainder of the pancreas was assayed for insulin content. Compared with baseline and control groups, PIO treatment significantly reduced blood glucose and glycosylated hemoglobin levels. Plasma insulin levels decreased significantly in ob/ob mice treated with PIO. All groups treated with PIO exhibited significantly greater beta-cell granulation, evidence of reduced beta-cell stress, and 1.5- to 15-fold higher levels of pancreatic insulin. The data from these studies suggest that comparable effects would be expected to slow the progression of type 2 diabetes, either delaying or possibly preventing progression to an insulin-dependent state. PMID- 14532172 TI - Overexpression of constitutively activated glutamate dehydrogenase induces insulin secretion through enhanced glutamate oxidation. AB - Glutamate dehydrogenase (GDH) catalyzes reversible oxidative deamination of l glutamate to alpha-ketoglutarate. Enzyme activity is regulated by several allosteric effectors. Recognition of a new form of hyperinsulinemic hypoglycemia, hyperinsulinism/hyperammonemia (HI/HA) syndrome, which is caused by gain-of function mutations in GDH, highlighted the importance of GDH in glucose homeostasis. GDH266C is a constitutively activated mutant enzyme we identified in a patient with HI/HA syndrome. By overexpressing GDH266C in MIN6 mouse insulinoma cells, we previously demonstrated unregulated elevation of GDH activity to render the cells responsive to glutamine in insulin secretion. Interestingly, at low glucose concentrations, basal insulin secretion was exaggerated in such cells. Herein, to clarify the role of GDH in the regulation of insulin secretion, we studied cellular glutamate metabolism using MIN6 cells overexpressing GDH266C (MIN6-GDH266C). Glutamine-stimulated insulin secretion was associated with increased glutamine oxidation and decreased intracellular glutamate content. Similarly, at 5 mmol/l glucose without glutamine, glutamine oxidation also increased, and glutamate content decreased with exaggerated insulin secretion. Glucose oxidation was not altered. Insulin secretion profiles from GDH266C overexpressing isolated rat pancreatic islets were similar to those from MIN6 GDH266C, suggesting observation in MIN6 cells to be relevant in native beta cells. These results demonstrate that, upon activation, GDH oxidizes glutamate to alpha-ketoglutarate, thereby stimulating insulin secretion by providing the TCA cycle with a substrate. No evidence was obtained supporting the hypothesis that activated GDH produced glutamate, a recently proposed second messenger of insulin secretion, by the reverse reaction, to stimulate insulin secretion. PMID- 14532173 TI - Nocardiosis: review of clinical and laboratory experience. PMID- 14532174 TI - Molecular subtyping of Vibrio cholerae O1 and O139 by pulsed-field gel electrophoresis in Hong Kong: correlation with epidemiological events from 1994 to 2002. AB - Two hundred twenty isolates of Vibrio cholerae O1 and O139 collected from 1994 to 2002 in Hong Kong were analyzed by pulsed-field gel electrophoresis (PFGE). Chromosomal DNAs from all V. cholerae isolates in agarose plugs were digested with the restriction enzyme NotI, resulting in 20 to 27 bands. Sixty distinctive PFGE patterns in the range of 10 to 300 kb were noted among 213 isolates typeable by PFGE. By comparing the common PFGE patterns obtained from four well-defined outbreaks of V. cholerae O1 and O139 with those obtained from other, epidemiologically unrelated isolates during the study period, indistinguishable and similar PFGE patterns were identified, indicating their close relatedness, in agreement with the results of epidemiological investigations. Heterogeneous PFGE patterns (with four to six banding differences), however, were identified among strains that were imported from other parts of Asia, including Indonesia, India, and Pakistan. Correlations with epidemiological information further support the usefulness of PFGE as an epidemiological tool in laboratory investigations of suspected outbreaks. Standardization of PFGE methodology will allow international comparison of fingerprint patterns and will form the basis of a laboratory network for tracking V. cholerae. PMID- 14532175 TI - Distribution of bovine herpesvirus type 5 DNA in the central nervous systems of latently, experimentally infected calves. AB - Bovine herpesvirus type 5 (BHV-5) is an alphaherpesvirus associated with meningoencephalitis, a disease highly prevalent in South America. In this study, we investigated the distribution of BHV-5 DNA in the brains of latently, experimentally infected calves by using a PCR for the glycoprotein B gene. Twelve calves inoculated intranasally with a Brazilian BHV-5 isolate were divided into two groups: group A calves (n = 4) were euthanized 55 days postinoculation (p.i.) for tissue collection; group B calves (n = 8) were submitted to dexamethasone administration at day 60 p.i. for reactivation of latent infection and were euthanized 50 days later. Latent infection was reactivated in all group B calves, as demonstrated by virus isolation from nasal secretions and/or seroconversion. Three calves developed neurological disease and died or were euthanized in extremis. For group A calves, viral DNA was consistently detected in the trigeminal ganglia (4/4), midbrain (4/4), thalamus (4/4), and olfactory cortex (4/4) and less frequently in the pons (3/4), cerebellum (3/4), anterior cerebral cortex (2/4), and olfactory bulb (2/4). For calves previously submitted to reactivation (group B), viral DNA was detected with roughly the same frequency in the same areas as for the group A calves. In addition, viral DNA was detected in the posterior (5/5) and dorso-lateral cortex (3/5). All DNA-positive tissues were negative for infectivity and viral antigens. These results demonstrated that latent BHV-5 DNA is present in several areas of the brain during latent infection and that virus reactivation may result in the establishment of latent infection in additional sites of the brain. PMID- 14532176 TI - Evaluation of reverse transcription-PCR assays for rapid diagnosis of severe acute respiratory syndrome associated with a novel coronavirus. AB - The reverse transcription (RT)-PCR protocols of two World Health Organization (WHO) severe acute respiratory syndrome (SARS) network laboratories (WHO SARS network laboratories at The University of Hong Kong [WHO-HKU] and at the Bernhard Nocht Institute in Hamburg, Germany [WHO-Hamburg]) were evaluated for rapid diagnosis of a novel coronavirus (CoV) associated with SARS in Hong Kong. A total of 303 clinical specimens were collected from 163 patients suspected to have SARS. The end point of both WHO-HKU and WHO-Hamburg RT-PCR assays was determined to be 0.1 50% tissue culture infective dose. Using seroconversion to CoV as the "gold standard" for SARS CoV diagnosis, WHO-HKU and WHO-Hamburg RT-PCR assays exhibited diagnostic sensitivities of 61 and 68% (nasopharyngeal aspirate specimens), 65 and 72% (throat swab specimens), 50 and 54% (urine specimens), and 58 and 63% (stool specimens), respectively, with an overall specificity of 100%. For patients confirmed to have SARS CoV and from whom two or more respiratory specimens were collected, testing the second specimen increased the sensitivity from 64 and 71% to 75 and 79% for the WHO-HKU and WHO-Hamburg RT-PCR assays, respectively. Testing more than one respiratory specimen will maximize the sensitivity of PCR assays for SARS CoV. PMID- 14532177 TI - Etiology of acute diarrhea in adults in southwestern Nigeria. AB - Stool specimens from 113 adult outpatients with diarrhea in southwestern Nigeria and 63 controls were examined for bacterial and parasitic enteric pathogens. Enterohemorrhagic Escherichia coli (EHEC) (P < 0.02), enteroaggregative E. coli (EAEC) (P < 0.02), and Entamoeba histolytica (P < 0.0002) were significantly associated with diarrhea. Salmonella, Shigella, nontoxigenic Vibrio cholerae, other categories of diarrheagenic E. coli, as well as a variety of helminths were recovered more frequently from the stools of patients than from the stools of controls but did not show a significant association with disease. Multiple pathogens were recovered from 36.3% of specimens, and bloody diarrhea was commonly associated with E. histolytica and diarrheagenic E. coli infections. The majority of EHEC isolates were non-O157 strains that carried the stx(2) gene. Of the 23 EHEC-infected patients, 12 (52.2%) presented during the 10th week of the study. EHEC strains isolated within this cluster were more likely to hybridize with the enterohemolysin gene probe, to be nonmotile and sorbitol positive, and to fail to agglutinate O157 antisera. Pulsed-field gel electrophoresis demonstrated that the only strains with XbaI profiles that occurred more than once were isolated during the 10th and 11th weeks of the study, suggesting an outbreak. The study has demonstrated that E. histolytica, EHEC, and EAEC are important diarrheal pathogens within the study area and that sporadic and epidemic EHEC infections occur in developing as well as developed countries. Routine surveillance for diarrheagenic E. coli, even only at the tertiary-care level, would be useful in identifying outbreaks and assist in identifying environmental reservoirs and transmission routes. PMID- 14532178 TI - New real-time reverse transcriptase-initiated PCR assay with single-copy sensitivity for human immunodeficiency virus type 1 RNA in plasma. AB - More sensitive assays for human immunodeficiency virus type 1 (HIV-1) RNA are needed to detect, quantify, and characterize persistent viremia in patients who are receiving antiretroviral therapy and whose plasma HIV-1 RNA levels are suppressed to less than 50 to 75 copies/ml. We therefore developed an internally controlled real-time reverse transcriptase-initiated PCR assay that quantifies HIV-1 RNA concentrations down to 1 copy per ml of plasma. This assay with single copy sensitivity (the single-copy assay) generates a reproducible linear regression plot of input copy number versus threshold cycle by using HIV-1 RNA transcripts at copy numbers ranging from 1 to 10(6) per reaction mixture. The single-copy assay was compared to the ultrasensitive AMPLICOR HIV-1 MONITOR assay and a more sensitive modification of the ultrasensitive assay by repeatedly testing a low-copy-number panel containing 200 to 0.781 copies of HIV-1 RNA per ml of plasma. This comparison showed that the single-copy assay had a greater sensitivity than the other assays and was the only assay that detected HIV-1 RNA at levels as low as 0.781 copies/ml. Testing of plasma samples from 15 patients who were receiving antiretroviral therapy and who had <75 HIV-1 RNA copies/ml revealed persistent viremia in all 15 patients, with HIV-1 RNA levels ranging from 1 to 32 copies/ml (median, 13 copies/ml). The greater sensitivity of the single-copy assay should allow better characterization of persistent viremia in patients who are receiving antiretroviral therapy and whose HIV-1 RNA levels are suppressed to below the detection limits of present assays. PMID- 14532179 TI - Madurella mycetomatis strains from mycetoma lesions in Sudanese patients are clonal. AB - Molecular diversity among clinical isolates of Madurella mycetomatis, the prime fungal agent of human mycetoma in Sudan, could possibly explain the diverse clinical presentations of this severely debilitating infectious disease. In addition, culture-independent DNA-mediated typing tests need to be developed for this organism, since M. mycetomatis DNA, but not the organism itself, can be identified in soil, the material from which infections are thought to originate. A collection of 38 different clinical M. mycetomatis isolates was characterized by large-scale random amplification of polymorphic DNA using 20 different primer species. These analyses, involving at least 2,600 annealing sites, showed a complete lack of DNA fingerprint variation among the various isolates. From the resulting homogeneous DNA fingerprints, seven fragments were cloned and sequenced, and novel, species-specific PCR restriction fragment length polymorphism (RFLP) tests were designed. The seven PCR RFLP tests were successfully performed on the 38 different M. mycetomatis strains. However, again all M. mycetomatis DNA patterns obtained appeared to be identical, whereas patterns produced using DNAs from other fungal species were clearly discriminatory. These results suggest that there is little genetic variation among clinically relevant M. mycetomatis strains from Sudan. The data tentatively imply that different manifestations of mycetoma are due to differences in host susceptibility rather than differential virulence of the causative agent. PMID- 14532180 TI - Molecular detection and identification of influenza viruses by oligonucleotide microarray hybridization. AB - Microarrays of virus-specific oligonucleotides may provide a method of screening samples for the presence or absence of a large variety of viruses simultaneously. Influenza viruses are ideal for evaluating such microarrays because of their genetic and host diversity, and the availability of an extensive sequence database. A collection of 476 influenza virus-specific oligonucleotides was spotted onto glass slides as probes. Viral RNAs were reverse transcribed and amplified by PCR, and the products were labeled with cyanine dyes. The presence of viruses and their identities were determined by hybridization. The fluorescence intensities of oligonucleotide spots were highly reproducible within each slide and satisfactorily proportional between experiments. However, the intensities of probe spots completely complementary to target sequences varied from background to saturation. The variations did not correlate with base composition, nucleotide sequence, or internal secondary structures. Therefore, thresholds for determining whether hybridization to a spot should be judged as positive were assigned individually. Considering only positive spots from probes predicted to be monospecific for influenza virus species, subtype, host source, or gene segment, this method made correct identifications at the species, hemagglutinin subtype, and gene segment levels. Monospecific neuraminidase (NA) subtype probes were insufficiently diverse to allow confident NA subtype assignment. Incorporating positive spots from polyspecific probes into the identification scheme gave similar results. Overall, the results demonstrate the potential of microarray-based oligonucleotide hybridization for multiple virus detection. PMID- 14532181 TI - Contribution of the Platelia Candida-specific antibody and antigen tests to early diagnosis of systemic Candida tropicalis infection in neutropenic adults. AB - The Platelia Candida-specific antigen and antibody assays (Bio-Rad Laboratories) were used to test serial serum samples from seven neutropenic adult patients with hematological malignancies who had developed systemic Candida tropicalis infections. The diagnosis of candidiasis was based on a positive blood culture (all seven patients) and the isolation of C. tropicalis from a normally sterile site (six patients). All patients received early antifungal therapy with amphotericin B and/or an azole derivative and had successful outcomes. When the combined assays were applied to sera collected at different time points before and after the first positive blood culture, all patients tested positive. In six patients, at least one positive test was obtained with sera collected, on average, 5 days (range, 2 to 10 days) prior to the first positive blood culture, while blood cultures were constantly negative. High and persistent mannanemias were detected in all patients during the neutropenic period. In five patients, an increased antibody response was detected when the patients recovered from aplasia. Controls consisted of 48 serum samples from 12 febrile neutropenic patients with aspergillosis (n = 4), bacteremia (n = 4), or no evidence of infection (n = 4). A low level of mannanemia was detected in only one serum sample, and none showed significant Candida antibody titers. Our data thus confirm the value of the combined detection of mannanemia and antimannan antibodies in individuals at risk of candidemia and suggest that in neutropenic patients, an approach based on the regular monitoring of both markers could contribute to the earlier diagnosis of C. tropicalis systemic infection. PMID- 14532182 TI - Clonal characterization of Staphylococcus aureus by multilocus restriction fragment typing, a rapid screening approach for molecular epidemiology. AB - We have developed a rapid and simplified approach for the strain characterization of Staphylococcus aureus on the basis of multilocus sequence typing (MLST) in which sequence variations in the MLST housekeeping gene loci are detected by restriction fragment pattern analysis rather than sequencing; we refer to this approach as multilocus restriction fragment typing (MLRFT). Briefly, MLRFT for S. aureus involves the PCR amplification of each of the seven MLST housekeeping gene loci by using the same primer pairs used in MLST. The amplicons are then digested directly with one or two restriction enzymes and the restriction fragments are resolved by agarose gel electrophoresis. Projection from published MLST data shows that MLRFT captures about 95% of the genetic diversity detected by MLST. The MLRFT approach was validated with a set of 59 methicillin-susceptible and 44 methicillin-resistant S. aureus isolates from community-acquired and nosocomial sources which had previously been characterized by pulsed-field gel electrophoresis (PFGE). MLRFT resolved the 103 isolates into 15 restriction fragment types, giving a discrimination index of 89.0%. Clonal groupings established by MLRFT correlated well with those established by PFGE. In short, MLRFT provides a convenient alternative to MLST and PFGE because it requires minimal laboratory facilities and is relatively simple and inexpensive to perform. PMID- 14532183 TI - Rapid diagnosis of mycobacterial infections and quantitation of Mycobacterium tuberculosis load by two real-time calibrated PCR assays. AB - Sensitive and specific techniques to detect and identify Mycobacterium tuberculosis directly in clinical specimens are important for the diagnosis and management of patients with tuberculosis (TB). We developed two real-time PCR assays, based on the IS6110 multicopy element and on the senX3-regX3 intergenic region, which provide a rapid method for the diagnosis of mycobacterial infections. The sensitivity and specificity of both assays were established by using purified DNA from 71 clinical isolates and 121 clinical samples collected from 83 patients, 20 of whom were affected by TB. Both assays are accurate, sensitive, and specific, showing a complementary pattern of Mycobacterium recognition: broader for the IS6110-based assay and restricted to the M. tuberculosis complex for the senX3-regX3-based assay. Moreover, the addition of a synthetic DNA calibrator prior to DNA extraction allowed us to measure the efficiency of DNA recovery and to control for the presence of PCR inhibitors. The mycobacterial burden of the clinical samples, as assessed by direct microscopy, correlates with the M. tuberculosis DNA load measured by the senX3-regX3-based assay. In addition, reduced levels of M. tuberculosis DNA load are present in those patients subjected to successful therapy, suggesting a potential use of this assay for monitoring treatment efficacy. Therefore, these assays represent a fully controlled high-throughput system for the evaluation of mycobacterial burden in clinical specimens. PMID- 14532184 TI - Fast and accurate quantitative detection of Helicobacter pylori and identification of clarithromycin resistance mutations in H. pylori isolates from gastric biopsy specimens by real-time PCR. AB - Rapid identification of patients infected with clarithromycin-resistant Helicobacter pylori without the need for culture can help to avoid useless prescriptions of clarithromycin. We developed and tested a routine real-time quantitative PCR assay dedicated to that purpose. One hundred ninety-six consecutive gastric biopsy specimens were examined by culture, histology performed by a trained physician, and rapid PCR with the LightCycler apparatus. Infection was defined as (i) positivity of culture, (ii) positivity of histology, or (iii) positivity of PCR if confirmed by positivity of a concomitant indirect test (serology or urea breath test). Susceptibility to clarithromycin was tested by E-test and PCR. The prevalence of infection was 33.7% (66 of 196 samples). The sensitivities of culture, histology, and PCR were 90.9% (60 of 66 samples), 87.9% (58 of 66 samples), and 97.0% (64 of 66 samples), respectively. The specificity of PCR was 94.6% (123 of 130 samples). The linearity of the PCR results was achieved over a 6-log range of input DNA, and we were able to accurately quantify as few as 300 bacteria and to qualitatively detect as few as 30 bacteria per DNA sample. For clarithromycin susceptibility testing, there was 98.2% (55 of 56 samples) concordance between E-test and PCR. Forty-eight strains were clarithromycin susceptible, and 9 strains were clarithromycin resistant. The single discrepancy concerned a culture which was a mixture of mutant and wild type, with a susceptible-to-resistant ratio of 11.5: the resistant population was detected by E-test but not by PCR. Our PCR assay is accurate for fast detection of H. pylori as well as of clarithromycin resistance and is also able to objectively determine bacterial density. PMID- 14532186 TI - Restriction fragment length polymorphism analysis of ribosomal DNA intergenic regions is useful for differentiating strains of Trichophyton mentagrophytes. AB - Twenty isolates of Tricophyton mentagrophytes var. mentagrophytes and 47 isolates of T. mentagrophytes var. interdigitale, identified by morphological characteristics, were screened by restriction fragment length polymorphism (RFLP) analysis of the PCR-amplified internal transcribed spacer (ITS) region of ribosomal DNA (rDNA). Sixty isolates (14 of 20 T. mentagrophytes var. mentagrophytes isolates and 46 of 47 T. mentagrophytes var. interdigitale isolates) shared an identical ITS RFLP profile and were further investigated by using a probe targeted to the rDNA nontranscribed spacer (NTS) region. Polymorphisms were observed in the NTS regions of both T. mentagrophytes var. mentagrophytes and T. mentagrophytes var. interdigitale isolates. Twenty-three individual RFLP patterns (DNA types P-1 to P-12 and A-1 to A-11) were recognized and divided into two groups depending on the presence (P) or absence (A) of a 2.5 kb band, which correlated to a large extent with the morphological variety. Eleven of 14 T. metagrophytes var. mentagrophytes isolates were A types, and all of the 46 T. mentagrophytes var. interdigitale isolates were P types. A majority of strains (23 of 60 [38.3%]) were characterized by one RFLP pattern (pattern P 1), and eight types (P-1 to P-6, P-8, and P-9) accounted for 75% (45 of 60) of all strains, including all of the T. mentagrophytes var. interdigitale isolates. The remaining 15 types were represented by one only isolate and included all of the T. mentagrophytes var. mentagrophytes isolates. We conclude that RFLP analysis of the rDNA NTS region is a valuable technique for differentiation of T. mentagrophytes strains. Furthermore, by use of this method, there appears to be a greater degree of diversity among T. mentagrophytes var. mentagrophytes isolates than among T. mentagrophytes var. interdigitale isolates. PMID- 14532185 TI - Association between handling of pet treats and infection with Salmonella enterica serotype newport expressing the AmpC beta-lactamase, CMY-2. AB - Resistance to the extended-spectrum cephalosporins can occur in Salmonella species via the production of extended-spectrum and AmpC beta-lactamases. We describe human infections with Salmonella enterica serotype Newport phage type 14 strains resistant to ceftazidime (CAZ) and cefoxitin (FOX) related to the handling of pet treats containing dried beef. These strains were isolated from five patients in Calgary, Alberta, Canada, during 2002 and were compared to a strain cultured from a commercial pet treat present at the property of one of the patients. The strains were resistant to FOX, CAZ, cefpodoxime, ampicillin, and chloramphenicol; intermediate resistant to ceftriaxone and cefotaxime; and sensitive to the aminoglycosides, ciprofloxacin, cefepime, and imipenem. Isoelectric focusing, multiplex PCR, and sequencing of the amplicons showed that all strains produced the plasmid-encoded AmpC beta-lactamase, CMY-2. Restriction analysis of plasmid DNA following transformation demonstrated that bla(CMY-2) was encoded on an approximately 140-kb plasmid. Pulsed-field gel electrophoresis showed the human and pet treat Salmonella strains to be highly related. This study is the first to implicate the transfer of multidrug-resistant Salmonella species through the handling of commercial pet treats containing animal products. In addition to documenting the first cases of human infection caused by CMY-2 producing S. enterica serotype Newport strains in Canada, this study illustrates the necessity of rapid and accurate laboratory-based surveillance in the identification of novel types of antimicrobial resistance. PMID- 14532187 TI - Evaluation of protocol using gene capture and PCR for detection of Helicobacter pylori DNA in feces. AB - The route of transmission of Helicobacter pylori, which is usually acquired in childhood and is one of the most common bacterial infections in humans, remains undetermined. Mapping the distribution of H. pylori genotypes within families could help to determine the routes of transmission and risk factors. Here we describe a noninvasive method for obtaining H. pylori DNA isolates from the feces of children. Children presenting with gastrointestinal symptoms at the Royal Hospital for Sick Children were tested for gastric H. pylori colonization by using the 13C-urea breath test (UBT) and were asked to provide fecal samples, which were tested for H. pylori by using the HpSA fecal antigen test. DNA was purified from fecal samples by using a novel method of gene capture with subsequent H. pylori PCR analysis. Fifteen UBT-positive and 15 UBT-negative children participated in the study. The positive and negative predictive values for the assay were 80 and 100%, respectively. Fecal DNA purification followed by H. pylori PCR analysis is an effective tool for harvesting H. pylori DNA isolates from the feces of children. This technique may be developed to allow the diagnosis and noninvasive genotyping of H. pylori in children and their families. PMID- 14532188 TI - Genome type analysis of adenovirus types 3 and 7 isolated during successive outbreaks of lower respiratory tract infections in children. AB - Adenovirus is an important cause of respiratory infections in infants and children. Fifty-one serotypes have been identified, and adenovirus type 3 (Ad3) and Ad7 have often been associated with outbreaks of severe respiratory tract infections. Each serotype can be further divided into genome types based on the patterns of digestion of their DNAs with restriction enzymes. DNA restriction analysis was performed with 56 strains of Ad3 and 98 strains of Ad7 by using 12 restriction enzymes recognizing 6 bp (BamHI, BclI, BglI, BglII, BstEII, EcoRI, HindIII, HpaI, SalI, SmaI, XbaI, and XhoI). The virus strains were isolated during outbreaks of lower respiratory tract infections in children during an 11 year period from 1990 to 2000 in Seoul, Korea. Among the Ad3 strains, seven genome types were identified; Ad3a and six novel types (Ad3a13, Ad3a14, Ad3a15, Ad3a16, Ad3a17, and Ad3a18). Multiple genome types cocirculated during outbreaks, and some of these were isolated during the 11-year observation period, while others were restricted to particular outbreaks. For Ad7, two genome types, Ad7d and Ad7l, the latter of which is a novel genome type, were identified. A shift in genome types occurred from Ad7d to Ad7l during successive outbreaks. Mortality was 3.6% among children with Ad3 infections and 18% among children infected with either of the Ad7 genome types. In conclusion, the data confirm that Ad3 genome types are more diverse than those of Ad7 and suggest that shifts of genome types may occur during successive outbreaks of Ad3 and Ad7. PMID- 14532189 TI - Segregation of human immunodeficiency virus type 1 subtypes by risk factor in Australia. AB - The aim of this study was to determine which human immunodeficiency virus type 1 (HIV-1) subtypes were circulating in Australia and to correlate the subtypes with risk factors associated with the acquisition of HIV-1 infection. DNA was extracted from peripheral blood mononuclear cells, and HIV-1 env genes were amplified and subtyped using heteroduplex mobility analysis, with selected samples sequenced and phylogenetic analysis performed. The HIV-1 env subtypes were determined for 141 samples, of which 40 were from female patients and 101 were from male patients; 13 samples were from children. Forty-seven patients were infected by homosexual or bisexual contact, 46 were infected through heterosexual contact, 21 were infected from injecting drug use (IDU), 13 were infected by vertical transmission, 8 were infected from nosocomial exposure, and 6 were infected by other modes of transmission, including exposure to blood products, ritualistic practices, and two cases of intrafamilial transmission. Five subtypes were detected; B (n = 104), A (n = 5), C (n = 17), E (CRF01_AE; n = 13), and G (n = 2). Subtype B predominated in HIV-1 acquired homosexually (94% of cases) and by IDU (100%), whereas non-subtype B infections were mostly seen in heterosexually (57%) or vertically (22%) acquired HIV-1 infections and were usually imported from Africa and Asia. Subtype B strains of group M viruses predominate in Australia in HIV-1 transmitted by homosexual or bisexual contact and IDU. However, non-B subtypes have been introduced, mostly acquired via heterosexual contact. PMID- 14532190 TI - Molecular evidence of persistent echovirus 13 meningoencephalitis in a patient with relapsed lymphoma after an outbreak of meningitis in 2000. AB - Enteroviral meningoencephalitis was diagnosed in a patient with an immunodeficiency syndrome acquired after treatment with rituximab for a relapsed primary B-cell lymphoma. A second meningoencephalitic episode was diagnosed 6 months later and was successfully treated with a combination of immunoglobulins and pleconaril. The infection was persistent since the enterovirus genome was detected in five sequential specimens of cerebrospinal fluid collected over 9 months. An echovirus 13 isolate was isolated in the first three samples. The viral sequence encoding the VP1 capsid protein of the three isolates was determined and was compared with that of four control viruses. The virus isolates recovered from the patient shared >99% nucleotide sequence similarity with one another. In a phylogenetic tree, they were directly related to a control virus obtained from a patient hospitalized in 2000 during an outbreak of enterovirus meningitis. The epidemiological origin of a chronic echovirus infection in a patient with immune deficiency suggests that the echovirus had been continuously circulating in the general population after the outbreak that had revealed its emergence. PMID- 14532191 TI - Effect of duplicate isolates of methicillin-susceptible and methicillin-resistant Staphylococcus aureus on antibiogram data. AB - Duplicate Staphylococcus aureus isolates were analyzed to determine the impact of multiple isolates from the same patient on annual antibiogram data. During a 6 year period (1996 to 2001), 3,227 patients with 4,844 S. aureus isolates were evaluated. A total of 39% of patients with methicillin-resistant S. aureus (MRSA) (n = 860) and 23% of patients with methicillin-susceptible S. aureus (MSSA) (n = 2,367) infections had duplicate isolates. Cumulative data show that 91% of the patients during this 6-year period with duplicate isolates (2 to 13 duplicates/year) did not switch between MSSA and MRSA but retained the original S. aureus strain whether it was MSSA or MRSA. Rates of MRSA were calculated for each year by using all isolates and then eliminating duplicates. The impact of duplicate MRSA and MSSA isolates was evaluated by using the ratio of isolates per patient such that ratios of >1.0 indicate >1 isolate per patient. The 6-year ratio for MRSA was 1.90 isolates/patient, and the ratio for MSSA was 1.35. A significant difference (P < 0.05) was noted in the MRSA rates in 4 of 6 years when duplicate isolates were removed. Common phenotypic antibiogram patterns were compared for all MRSA isolates during the 6-year period, and 64% were of a single antibiogram phenotype. Eighty-eight percent of patients with duplicate MRSA isolates had phenotypically identical multiple isolates. The rate of MRSA differs when duplicate isolates are removed from the antibiogram data. PMID- 14532192 TI - Predominance of Ehrlichia ewingii in Missouri dogs. AB - To investigate the species distribution of Ehrlichia present in Missouri dogs, we tested 78 dogs suspected of having acute ehrlichiosis and 10 healthy dogs. Blood from each dog was screened with a broad-range 16S rRNA gene PCR assay that detects known pathogenic species of Ehrlichia and ANAPLASMA: The species was determined by using species-specific PCR assays and nucleotide sequencing. Ehrlichia antibody testing was performed by using an indirect immunofluorescence assay with Ehrlichia chaffeensis as the antigenic substrate. The broad-range assay detected Ehrlichia or Anaplasma DNA in 20 (26%) of the symptomatic dogs and 2 (20%) of the asymptomatic dogs. E. ewingii accounted for 20 (91%), and E. chaffeensis accounted for 1 (5%) of the positives. Anaplasma phagocytophilum DNA was detected in one dog, and the sequences of regions of the 16S rRNA gene and the groESL operon amplified from the blood of this dog matched the published sequences of this organism. Antibodies reactive with E. chaffeensis were detected in 14 (67%) of the 21 PCR-positive dogs and in 12 (19%) of the 64 PCR-negative dogs. Combining the results of PCR and serology indicated that 33 (39%) of 85 evaluable dogs had evidence of past or current Ehrlichia infection. We conclude that E. ewingii is the predominant etiologic agent of canine ehrlichiosis in the areas of Missouri included in this survey. E. canis, a widely recognized agent of canine ehrlichiosis, was not detected in any animal. The finding of E. ewingii in asymptomatic dogs suggests that dogs could be a reservoir for this Ehrlichia species. PMID- 14532193 TI - Evaluation of the Hodge test and the imipenem-EDTA double-disk synergy test for differentiating metallo-beta-lactamase-producing isolates of Pseudomonas spp. and Acinetobacter spp. AB - Gram-negative bacilli with acquired metallo-beta-lactamase (MBL) production have been increasingly reported in some countries, necessitating their detection. The aim of this study was to evaluate the performance of the Hodge test and those of the imipenem (IPM)-EDTA, ceftazidime (CAZ)-mercaptopropionic acid (MPA), and CAZ sodium mercaptoacetic acid (SMA) double-disk synergy tests (DDSTs). The efficiencies of testing CAZ-resistant and IPM-nonsusceptible isolates were also compared. Strains used for the evaluation were known IMP-1 and VIM-2 MBL producing isolates and consecutive and CAZ-nonsusceptible isolates of pseudomonads and acinetobacters. The performance of the Hodge test was improved by addition of zinc sulfate (140 microg/disk) to an IPM disk. In DDSTs, EDTA (ca. 1,900 microg) disks were better at detecting MBL-producing strains among pseudomonads, while MPA (3 microl) and SMA (3 mg) disks performed better for acinetobacters. EDTA (ca. 750 microg)-plus-SMA (ca. 2 mg) disks performed better than EDTA, MPA, or SMA disks with both organisms. CAZ-SMA DDSTs failed to detect 22 of 80 (28%) MBL-producing acinetobacters. In conclusion, use of an IPM disk and an EDTA (750 microg)-plus-SMA (2 mg) disk improves performance, and testing IPM-nonsusceptible isolates rather than CAZ-resistant isolates could reduce screening work. Further evaluation of the test is required for the detection of other types of MBL-producing gram-negative bacilli. PMID- 14532194 TI - Detection of a point mutation associated with high-level isoniazid resistance in Mycobacterium tuberculosis by using real-time PCR technology with 3'-minor groove binder-DNA probes. AB - Tuberculosis remains one of the leading infectious causes of death worldwide. The emergence of drug-resistant strains of Mycobacterium tuberculosis is a serious public health threat. Resistance to isoniazid (INH) is the most prevalent form of resistance in M. tuberculosis and is mainly caused by mutations in the catalase peroxidase gene (katG). Among high-level INH-resistant isolates (MIC > or = 2), 89% are associated with a mutation at codon 315 of katG. There is a need to develop rapid diagnostic tests to permit appropriate antibiotic treatment and to improve clinical management. Therefore, a single-tube real-time PCR, using a novel kind of probe (3'-minor groove binder-DNA probe), was developed to detect either the wild-type or the mutant codon directly in Ziehl-Neelsen-positive sputum samples. The detection limit of the assay for purified DNA was 5 fg per well (one mycobacterial genome), and with spiked sputum samples, it was 20 copies per well, corresponding to 10(3) mycobacteria per ml of sputum. Sputum samples from 20 patients living in Kazakhstan or Moldova and infected with monodrug- or multidrug-resistant M. tuberculosis and 20 sputum samples from patients infected with INH-susceptible M. tuberculosis were tested. The sensitivities and specificities of the probes were 70 and 94% for the wild-type probe and 82 and 100% for the mutant probe. Binding to either probe was nonambiguous. This real time PCR allows the rapid identification of a mutant katG allele and can easily be implemented in a clinical microbiology laboratory. PMID- 14532195 TI - Multiplexed, real-time PCR for quantitative detection of human adenovirus. AB - Adenovirus infection is becoming increasingly recognized as a cause of morbidity and mortality in the immunosuppressed patient population. While early detection and quantitation of adenovirus in peripheral blood has been suggested as a means of directing and monitoring antiviral therapy in these patients, few methods have been published, particularly with respect to viral quantitation. A multiplexed real-time PCR assay was developed that can quantitatively detect a wide range of known serotypes of human adenovirus, including all of subgroups A to C. This assay was compared to a qualitative, Southern blot-based PCR assay by using 45 peripheral blood specimens from 16 patients. There was 100% concordance between the two tests in terms of qualitative results. The real-time assay detected adenovirus in patient samples at levels from <200 to 266,681 copies/ml of blood. By using control viral samples, sensitivity was demonstrated to less than 10 copies of viral genome per reaction and quantitative linearity was demonstrated from 10 to 10(6) copies of input viral DNA. Equivalent sensitivity and linearity were demonstrated for 15 different reference serotypes of adenovirus. Eleven other viral serotypes have complete target region sequence homology to one or more of the strains tested. No cross-reactivity was noted with other commonly isolated viral species. Sequence analysis showed no significant homology with any other human pathogens (bacterial or viral). This assay allows rapid, sensitive, and specific quantitation of adenovirus and may have a significant impact on the care of immunocompromised patients at risk for disseminated viral infection. PMID- 14532196 TI - Human metapneumovirus infection in the Canadian population. AB - Human metapneumovirus (hMPV), a newly discovered paramyxovirus, has been associated with acute respiratory tract infections (ARIs) ranging from upper ARIs to severe bronchiolitis and pneumonia. Important questions remain on the contribution of hMPV to ARIs and its impact on public health. During the 2001 2002 season, we conducted a collaborative study with four provincial public health laboratories to study the prevalence of this new virus in the Canadian population. A total of 445 specimens were collected from patients of all age groups with ARIs and were tested for the presence of hMPV by reverse transcription-PCR. Of these, 66 (14.8%) tested positive for hMPV. Positive specimens were found in all age groups and in all four provinces studied. Virus activity peaked in February and March. The age range of the patients with hMPV infection was 2 months to 93 years (median age, 25 years), with similar numbers of females (35%) and males (41%). Thirty-three percent (n = 22) of hMPV-infected patients were hospitalized; of these, 27% (n = 6) had rhinitis and pneumonia, 23% (n = 5) had bronchiolitis, and 9% (n = 2) had bronchitis. The hospitalization rates were significantly higher among patients <5 years of age (P = 0.0005) and those >50 years of age (P = 0.0044) than among those 6 to 50 years of age. Phylogenetic analysis of the F gene showed that two hMPV genetic clusters were cocirculating in the 2001-2002 season, and comparison with earlier studies suggests a temporal evolutionary pattern of hMPV isolates. These results provide further evidence of the importance of hMPV in ARIs, particularly in young children and elderly individuals. PMID- 14532197 TI - Use of 16S rRNA gene sequencing for rapid identification and differentiation of Burkholderia pseudomallei and B. mallei. AB - Burkholderia pseudomallei and B. mallei, the causative agents of melioidosis and glanders, respectively, are designated category B biothreat agents. Current methods for identifying these organisms rely on their phenotypic characteristics and an extensive set of biochemical reactions. We evaluated the use of 16S rRNA gene sequencing to rapidly identify these two species and differentiate them from each other as well as from closely related species and genera such as Pandoraea spp., Ralstonia spp., Burkholderia gladioli, Burkholderia cepacia, Burkholderia thailandensis, and Pseudomonas aeruginosa. We sequenced the 1.5-kb 16S rRNA gene of 56 B. pseudomallei and 23 B. mallei isolates selected to represent a wide range of temporal, geographic, and origin diversity. Among all 79 isolates, a total of 11 16S types were found based on eight positions of difference. Nine 16S types were identified in B. pseudomallei isolates based on six positions of difference, with differences ranging from 0.5 to 1.5 bp. Twenty-two of 23 B. mallei isolates showed 16S rRNA gene sequence identity and were designated 16S type 10, whereas the remaining isolate was designated type 11. This report provides a basis for rapidly identifying and differentiating B. pseudomallei and B. mallei by molecular methods. PMID- 14532198 TI - emm typing of M nontypeable invasive group A streptococcal isolates in Israel. AB - We performed emm typing of M nontypeable invasive group A streptococcal (GAS) isolates collected in a prospective population-based study in Israel. One hundred twenty of 131 isolates (92%) had emm sequences compatible with GAS, consisting of 51 different emm types. Eleven isolates were found to be group G streptococcus. Of the 120 isolates, 55 (46%) belonged to 32 types for which there were no typing sera available in the Streptococcal Reference Laboratory in Israel. The other 65 (64%) isolates, consisting of 19 types, had sera available and therefore could have been serotyped. Forty-three isolates had T and emm types which were not correlated according to standard M-typing protocols and were therefore missed. The principal effect of emm typing was the addition of 32 types not previously identified in Israel and the discovery of new associations between emm and T types. emm typing did not significantly change the proportion of M types; the five most common types were 3, 28, 2, 62, and 41. Twenty different types comprised 80% of all isolates. No new emm sequences were discovered. emm typing emphasized the unusually low incidence of M1 strains causing severe disease in Israel. As serological typing of GAS becomes more problematic due to lack of sera and the appearance of new emm types, reference laboratories should replace M typing with emm sequence typing. Development of a GAS vaccine relies on the emm type distributions in different geographical locations. In our study, 7% of isolates (types 41 and 62) are not included in a 26-valent vaccine that is being studied. PMID- 14532199 TI - Decreased vancomycin susceptibility of coagulase-negative staphylococci in a neonatal intensive care unit: evidence of spread of Staphylococcus warneri. AB - Coagulase-negative staphylococci (CoNS) are important pathogens in premature neonates; decreasing glycopeptide susceptibility has been observed among these isolates. The epidemiology of colonization with CoNS, the organisms' vancomycin susceptibilities, and genetic relatedness were studied over 6 months in a tertiary-care neonatal unit. A total of 321 isolates of CoNS were isolated. Seventy-five percent of the infants were colonized at admission, and virtually all were colonized thereafter. Common species were Staphylococcus epidermidis (69%), S. warneri (12%), S. haemolyticus (9.7%), and S. hominis (5.6%). A total of 3.9% of CoNS isolates had decreased vancomycin susceptibility (DVS) (MICs > 2.0 microg/ml); isolate recovery was associated with a stay in a neonatal intensive care unit for >28 days (P = 0.039), vancomycin exposure (P = 0.021), and S. warneri colonization (P < 0.0001). Nine of 12 (75%) CoNS with DVS were S. warneri, had enhanceable high-level resistance in vitro, were indistinguishable or closely related by pulsed-field gel electrophoresis, and were different from 29 vancomycin-susceptible S. warneri isolates. Epidemiological analysis suggested unsuspected nosocomial spread. Species determination in certain settings may aid in the understanding of emerging nosocomial problems. PMID- 14532200 TI - Prediction of decreased susceptibility to penicillin of Neisseria meningitidis strains by real-time PCR. AB - Sequence analysis of the penA gene, encoding penicillin-binding protein 2 (PBP2), in 30 penicillin-intermediate (PenI) Neisseria meningitidis strains showed altered gene sequences due to the translocation of exogenous DNA blocks derived from commensal neisseriae, which are known to have PBP2 proteins with decreased affinity for the antibiotic. In order to obtain a rapid and reproducible method for predicting the PenI phenotype, a real-time PCR assay was set up with primers and probes designed on the basis of the penA gene. The A-->G mutation at codon 566, in the transpeptidase domain of the penA gene (which is present in the whole sample of 30 PenI strains and in all the 41 sequences of PenI meningococci isolated worldwide and has been deposited in the sequence databank), was chosen as a marker of penA translocations. Two hybridization probes were designed to distinguish the wild-type penA gene in penicillin-susceptible (PenS) meningococci from the mutated penA gene at codon 566 in PenI strains. Thermal analysis of probe hybridization revealed a melting temperature difference of at least 6 degrees C between PenI and PenS strains. This real-time PCR protocol characterizes the penicillin phenotype of N. meningitidis in a few hours without DNA sequencing and is useful for rapid screening of the penicillin-intermediate genotype among meningococcal isolates. PMID- 14532201 TI - Comparison of a shiga toxin enzyme-linked immunosorbent assay and two types of PCR for detection of shiga toxin-producing Escherichia coli in human stool specimens. AB - Shiga toxin (Stx)-producing Escherichia coli (STEC) is a major cause of sporadic cases of disease as well as serious outbreaks worldwide. The spectrum of illnesses includes mild nonbloody diarrhea, hemorrhagic colitis, and hemolytic uremic syndrome. STEC produces one or more Stxs, which are subdivided into two major classes, Stx1 and Stx2. The ingestion of contaminated food or water, person to-person spread, and contact with animals are the major transmission modes. The infective dose of STEC may be less than 100 organisms. Effective prevention of infection is dependent on rapid detection of the causative bacterial pathogen. In the present study, we examined 295 stool specimens for the presence of Stx producing E. coli by three different methods: an Stx enzyme-linked immunosorbent assay, a conventional PCR assay, and a LightCycler PCR (LC-PCR) assay protocol recently developed by our laboratory at the Institute of Medical Microbiology at Hannover Medical School. Our intent was to compare these three methods and to examine the utility of the STEC LC-PCR protocol in a clinical laboratory. The addition of a control DNA to each sample to clearly discriminate inhibited specimens from negative ones enhanced the accuracy of the LC-PCR protocol. From our results, it can be concluded that LC-PCR is a very useful tool for the rapid and safe detection of STEC in clinical samples. PMID- 14532202 TI - Genotypic analyses of Vibrio parahaemolyticus and development of a pandemic group specific multiplex PCR assay. AB - A total of 54 Vibrio parahaemolyticus strains including pandemic O3:K6 strains and newly emerged O4:K68, O1:K25, O1:K26, and O1:K untypeable strains (collectively referred to as the "pandemic group") were examined for their pulsed field gel electrophoresis (PFGE) and arbitrarily primed PCR (AP-PCR) profiles and for the presence or absence of genetic marker DNA sequences, toxRS/new or orf8, that had been reported elsewhere to be specific for the pandemic group. Both PFGE and AP-PCR analyses indicated that all strains of the pandemic group formed a distinct genotypic cluster, suggesting that they originated from the same clone. In addition to the pandemic group, four O3:K6 strains that did not possess the thermostable direct hemolysin (tdh) gene also belonged to this cluster and possessed the toxRS/new sequence. However, three O3:K6 strains that clearly belonged to the pandemic group by PFGE and AP-PCR did not possess the orf8 sequence. The evidence suggests that neither the toxRS/new nor the orf8 sequence is a reliable gene marker for definite identification of the pandemic group. We therefore developed a novel multiplex PCR assay specific for the pandemic group. The assay successfully distinguished pandemic group strains from other V. parahaemolyticus strains by yielding two distinct PCR products for tdh (263 bp) and the toxRS/new sequence (651 bp). PMID- 14532203 TI - Detection of Staphylococcus aureus enterotoxins A to D by real-time fluorescence PCR assay. AB - Staphylococcus aureus is one of the most significant pathogens causing nosocomial and community-acquired infections. Among the secreted staphylococcal virulence factors, there is a growing list of enterotoxins which can induce gastroenteric syndrome and toxic shock syndrome. Here, we developed a real-time fluorescence PCR assay (TaqMan PCR) for the detection of genes encoding staphylococcal enterotoxins A, B, C1, and D (SEA, SEB, SEC1, and SED) of S. aureus as well as the mecA gene encoding methicillin resistance and the femB gene as a specific genomic marker for S. aureus. SEA to SED were selected because they are the four classically described enterotoxins of S. aureus and because they were detected by latex agglutination. In order to evaluate the reliability of TaqMan PCR, we investigated 93 isolates of S. aureus derived from patients at our hospital over 5 months and compared the results with data obtained by a commercially available reversed passive latex agglutination assay (SET-RPLA) for these isolates. Thirteen enterotoxin genes were detected by TaqMan PCR; however, no proteins expressed by these genes were detected by SET-RPLA. As a result, more isolates of S. aureus (n = 44) were found positive by TaqMan PCR for one or more enterotoxin genes than by SET-RPLA for the respective proteins expressed by these genes (n = 40). We conclude that TaqMan PCR is more sensitive because it offers the possibility for determining enterotoxins on a genotypic basis. Additionally, the assay allows the parallel detection of genes for SEA to SED and methicillin resistance in S. aureus. Furthermore, real-time PCR is well suited for screening large numbers of samples at the same time, allowing rapid, reliable, efficient, and cost-saving routine laboratory diagnosis. PMID- 14532204 TI - Six-month multicenter study on invasive infections due to group B streptococci in Argentina. AB - There is little information about invasive infections by group B streptococci (GBS) and their antimicrobial susceptibilities in Latin America. We performed a prospective multicenter study to determine the serotype distribution and the antimicrobial susceptibility of GBS in Argentina. We identified 58 cases, but only 44 had sufficient data to be evaluated. Eight early-, four late-, and one fatal late, late-onset neonatal infections due to GBS were found. A total of 31 patients were adults with bacteremia, skin and soft tissue infections, osteomyelitis, arthritis, meningitis, abdominal infections, and renal abscess. Serotype III was prevalent in late-onset neonatal disease, and several serotypes (Ia/c, III, Ia, and II) were involved in early-onset neonatal infections. Serotypes II, Ia/c, III, and IV were commonly found in adults, with serotype II prevalent in younger adults (18 to 69 years old) and serotype Ia/c prevalent in elderly adults (>70 years old). The mortality rate attributable to GBS infections was 10.8%. All GBS were susceptible to penicillin and ceftriaxone. Resistance to clindamycin (1.7%), erythromycin (5.2%), azithromycin (5.2%), minocycline (69%), and tetracycline (72.4%), to high levels of kanamycin and amikacin (1.7%), and to intermediately high levels of gentamicin (1.7%) was observed. The bifunctional enzyme AAC6'-APH2" was detected in the isolate resistant to aminoglycosides, and other genetic determinants were identified in other resistant isolates: tetM and tetO in tetracycline-resistant streptococci and mefA and ermTR for efflux mediated and inducible macrolide-lincosamide-streptogramin B-resistant streptococci, respectively. For clinical purposes and rapid and easy detection of high-level aminoglycoside-resistant GBS, a screening method that used 1,000- micro g kanamycin disks is proposed. PMID- 14532205 TI - Description of a new yeast species, Malassezia japonica, and its detection in patients with atopic dermatitis and healthy subjects. AB - Lipophilic yeasts of the genus Malassezia are part of the normal cutaneous microflora and are considered one of the factors that trigger atopic dermatitis (AD). We isolated two strains of Malassezia from a healthy Japanese female. Analysis of the D1/D2 26S ribosomal DNA and internal transcribed spacer region sequences of the isolates suggested that they are new members of the genus Malassezia. We propose the name Malassezia japonica sp. nov. for the isolates. M. japonica is easily distinguished from the seven known lipophilic species by its ability to assimilate Tween 40 and Tween 60 and its inability to assimilate Tween 20 and Tween 80 and to grow at 40 degrees C. Furthermore, by applying transparent dressings to the skin lesions of 36 patients with AD and the skin of 22 healthy subjects, M. japonica DNA was detected by a non-culture-based method consisting of nested PCR with M. japonica species-specific primers. M. japonica DNA was detected from 12 of the 36 patients (33.3%) and 3 of the 22 healthy subjects (13.6%). Although it is not known whether M. japonica plays a role in AD, this species was part of the microflora in both patients with AD and healthy subjects. PMID- 14532206 TI - Diagnosis of Helicobacter pylori infection in adults with intellectual disability. AB - Helicobacter pylori infection is common among adults with intellectual disability. The acceptabilities and accuracies of different diagnostic tests in this population are unknown. We aimed to determine (i) patient acceptability and (ii) performance characteristics of serology, fecal-antigen, and urea breath tests among adults with intellectual disability. One hundred sixty-eight such adults underwent H. pylori testing with serology and fecal-antigen tests, and a portion underwent treatment. One year later, the participants were retested with fecal-antigen, serology, and urea breath tests. The numbers of specimens obtained and difficulties in collection reported by caregivers were noted. Test performance characteristics were assessed among participants and 65 of their caregivers, using serology as the reference. All participants provided at least one specimen, despite reported collection difficulties for 23% of fecal and 27% of blood specimens. Only 25% of the participants provided breath specimens; failure to perform this test was associated with lower intellectual ability and higher maladaptive behavior. The sensitivity, specificity, and positive and negative predictive values of the fecal test (baseline and 12 months versus caregivers) were 70 and 63 versus 81, 93 and 95 versus 98, 96 and 92 versus 93, and 53 and 74 versus 93%, respectively; those of the urea breath test (12 months versus caregivers) were 86 versus 100, 88 versus 95, 75 versus 89, and 94 versus 100%, respectively. With assistance, fecal or blood specimens for H. pylori assessment can be provided by most patients with intellectual disability regardless of their level of function or behavior. Only those with greater ability can perform the urea breath test. Using serology as the reference test, the limitations of performance characteristics of the fecal-antigen and urea breath tests are similar to those among a control group of caregivers. PMID- 14532207 TI - Evaluation of the VITEK 2 system for rapid direct identification and susceptibility testing of gram-negative bacilli from positive blood cultures. AB - This study explores the possibility of combining the BacT/Alert Microbial Detection System with the VITEK 2 system to achieve rapid bacterial identification and susceptibility testing. Direct inoculation of bacterial suspension to the VITEK 2 ID-GNB card and AST-NO09 card was made by differential centrifugation of blood cultures of organisms with gram-negative enteric bacillus like morphology. A total of 118 strains were investigated; of these, 97 (82.2%) strains were correctly identified to the species level and 21 (17.8%) strains were not identified; by comparing the results with those of the reference method of API identification systems using a pure culture, it was found that no strain had been misidentified. Among the 21 strains with no identification, 13 (61.9%) strains were nonfermenters. The direct-identification reporting time of VITEK 2 was 3.3 h. Direct testing of susceptibility to 11 antibiotics, i.e., amikacin, cefepime, ceftazidime, ciprofloxacin, gentamicin, imipenem, meropenem, netilmicin, piperacillin, piperacillin-tazobactam, and tobramycin, was also performed by using the broth microdilution (MB) method according to the NCCLS guidelines as a reference. After comparing the MICs of the VITEK 2 system with those obtained by the MB method within +/-twofold dilution, it was determined that the 1,067 organism-antibiotic combinations had an overall correct rate of 97.6% (1,041 combinations). The rates of susceptibility to the 11 antibiotics ranged from 88.7 to 100%, respectively. Only two (0.2%) and four (0.4%) combinations of the susceptibility tests gave very major errors (i.e., reported as sensitive by the VITEK 2 system but shown to be resistant by the MB method) and major errors (i.e., reported as resistant by the VITEK 2 system but shown to be sensitive by the MB method), respectively. The reporting time for the direct testing of susceptibility against the 11 antibiotics for 97 blood culture isolates by the VITEK 2 system ranged from 3.3 to 17.5 h. Compared with conventional methods that require 1 or 2 days, this method can make same-day reporting possible and thus permit better patient management. PMID- 14532208 TI - Algorithm to determine cost savings of targeting antimicrobial therapy based on results of rapid diagnostic testing. AB - A rapid diagnosis of pneumococcal pneumonia may allow the earlier use of narrow spectrum antimicrobial therapy. It is unknown, however, whether rapid diagnostic testing of patients hospitalized with community-acquired pneumonia (CAP) admitted to hospital lowers costs. Therefore, an algorithm to calculate the costs associated with the diagnosis and treatment of CAP was formulated. Subsequently, the algorithm was applied to clinical data for 122 consecutively admitted patients with CAP whose sputum samples were Gram stained and whose urine was tested for Streptococcus pneumoniae antigen. The costs of initial antimicrobial therapy, personnel, and materials were measured. Compared to the most expensive empirical regimen, rapid diagnostic testing would result in cost savings per patient (PP) of 3.51 Euros for Gram staining and 8.11 Euros for urinary pneumococcal antigen testing (1 Euro is equal to 1.13 US dollars, from 2000 to 2002). Compared to the cheapest regimen, Gram staining would increase the cost by 2.25 Euros PP, and urinary antigen testing would increase the cost by 24.26 Euros PP. In our setting, the use of rapid diagnostic testing would not lower costs. Cost savings depend, however, on the differences in the prices of the different antibiotics chosen and the proportion of evaluable and positive samples. PMID- 14532209 TI - Detection of simulated candidemia by the BACTEC 9240 system with plus aerobic/F and anaerobic/F blood culture bottles. AB - We studied the ability of the BACTEC 9240 automated blood culture system to detect simulated candidemia, including both Candida albicans and non-albicans Candida species. Simulated blood cultures were produced using 50 Candida isolates and BACTEC Plus Aerobic/F and Anaerobic/F blood culture bottles. Ten milliliters of blood and a suspension of each isolate containing 1,000 CFU were introduced into each bottle and then incubated at 35 degrees C in the BACTEC 9240 system. The system detected growth in 56 of 100 bottles. Four isolates did not have growth detected in either bottle after 21 days of incubation, resulting in four missed episodes of candidemia. If the blood culture bottles had been incubated for 5 days, an additional episode of candidemia would have remained undetected. If the bottles had been incubated for only 3 days, another episode would have been missed, resulting in up to six missed episodes of candidemia (four Candida glabrata isolates, one C. albicans isolate, and one Candida rugosa isolate). Terminal subculture of bottles without detected growth recovered yeast in 93% (41 of 44) of the bottles, representing 41 false negatives. In bottles where growth was detected, the time to detection was approximately 24 h. However, the mean time to growth detection for C. glabrata isolates in anaerobic medium was 22.14 +/- 2.47 h, but it was 120.89 +/- 35.33 h in aerobic medium (P < 0.001). The BACTEC 9240 system detected growth of most Candida isolates; however, the delayed time to detection of C. glabrata is clinically significant. Given the high rate of false negatives, terminal subcultures may be helpful in certain situations. PMID- 14532210 TI - Use of turbidimetric growth curves for early determination of antifungal drug resistance of filamentous fungi. AB - A previously described microbroth kinetic system (J. Meletiadis, J. F. Meis, J. W. Mouton, and P. E. Verweij, J. Clin. Microbiol. 39:478-484, 2001) based on continuous monitoring of changes in the optical density of fungal growth was used to describe turbidimetric growth curves of different filamentous fungi in the presence of increasing concentrations of antifungal drugs. Therefore, 24 clinical mold isolates, including Rhizopus oryzae, Aspergillus fumigatus, Aspergillus flavus, and Scedosporium prolificans, were tested against itraconazole, terbinafine, and amphotericin B according to NCCLS guidelines. Among various parameters of the growth curves, the duration of the lag phase was strongly affected by the presence of antifungal drugs. Exposure to increasing drug concentrations resulted in prolonged lag phases of the turbidimetric growth curves. The lag phases of the growth curves at drug concentrations which resulted in more than 50% growth (for itraconazole and terbinafine) and more than 75% growth (for amphotericin B) after 24 h of incubation for R. oryzae, 48 h for Aspergillus spp., and 72 h for S. prolificans were 4 h longer than the lag phases of the growth curves at the corresponding drug-free growth controls which varied from 4.4 h for R. oryzae, 6.5 h for A. flavus, 7.9 h for A. fumigatus, and 11.6 h for S. prolificans. The duration of the lag phases showed small experimental and interstrain variability, with differences of less than 2 h in most of the cases. Using this system, itraconazole and terbinafine resistance (presence of >50% growth) as well as amphotericin B resistance (presence of >75% growth) was determined within incubation periods of 5.0 to 7.7 h for R. oryzae (for amphotericin B resistance incubation for up to 12 h was required), 8.8 to 11.4 h for A. fumigatus, 6.7 to 8.5 h for A. flavus, and 13 to 15.6 h for S. prolificans while awaiting formal MIC determination by the NCCLS reference method. PMID- 14532211 TI - Extended epidemic of nosocomial urinary tract infections caused by Serratia marcescens. AB - In recent years a significant increase in the incidence of Serratia marcescens infections was noted at the Chang Gung Memorial Hospital, Taoyuan, Taiwan. A review of laboratory (1991 to 2002) and infection control (1995 to 2002) records showed the possibility of an extended epidemic of nosocomial urinary tract infections (UTIs) caused by S. marcescens. Therefore, in 1998 and 1999, 87 isolates were collected from patients with such infections and examined and another 51 isolates were collected in 2001 and 2002. The patients were mostly elderly or the infections were associated with the use of several invasive devices. S. marcescens was usually the only pathogen found in urine cultures in our study. Neither prior infections nor disseminated infections with the organism were observed in these patients. Resistance to most antibiotics except imipenem was noted. Two genotyping methods, pulsed-field gel electrophoresis and infrequent-restriction-site PCR, were used to examine the isolates. A total of 12 genotypes were identified, and 2 predominant genotypes were found in 72 (82.8%) of the 87 isolates derived from all over the hospital. However, 63.9% of the isolates of the two genotypes were from neurology wards. A subsequent intervention by infection control personnel reduced the infection rate greatly. The number and proportion of the two predominant genotypes were significantly reduced among the 51 isolates collected in 2001 and 2002. Thus, a chronic and long-lasting epidemic of nosocomial UTIs caused by S. marcescens was identified and a successful intervention was carried out. Both a cautious review of laboratory and infection control data and an efficient genotyping system are necessary to identify such a cryptic epidemic and further contribute to the quality of patient care. PMID- 14532212 TI - Utility of multilocus sequence typing as an epidemiological tool for investigation of outbreaks of gastroenteritis caused by Campylobacter jejuni. AB - Multilocus sequence typing (MLST) has been proven useful for the study of the global population structure of Campylobacter jejuni; however, its usefulness for the investigation of outbreaks of disease caused by C. jejuni has not been proven. In this study, MLST plus sequencing of the flaA short variable region (SVR) were applied to 47 isolates from 12 outbreaks of C. jejuni infection whose relatedness has been determined previously, and the results were compared to those of serotyping and pulsed-field gel electrophoresis (PFGE). Isolates implicated in an outbreak were indistinguishable by all four subtyping methods, with sporadic isolates being distinguished from outbreak isolates. Two sporadic isolates from one outbreak were resistant to SmaI digestion and therefore nontypeable by PFGE but were differentiated from the outbreak strain by the other methods. PFGE and flaA SVR typing were the most discriminatory methods, with discriminatory indices (DI) of 0.930 and 0.923, respectively. However, an epidemic strain from one outbreak was distinguished from the other outbreak isolates by flaA SVR typing; its flaA allele was different at five nucleotides, suggesting that this change was possibly mediated by recombination. MLST was less discriminatory than PFGE and flaA SVR typing (DI = 0.859), and many of the epidemic strains possessed common sequence types (STs) including ST-8, -21, -22, and -42. However, further discrimination within STs was achieved by flaA SVR typing or PFGE. The results from this study demonstrate that a combined approach of MLST plus flaA SVR typing provides a level of discrimination equivalent to PFGE for outbreak investigations. PMID- 14532213 TI - Practical disk diffusion method for detection of inducible clindamycin resistance in Staphylococcus aureus and coagulase-negative staphylococci. AB - Resistance to macrolides in staphylococci may be due to active efflux (encoded by msrA) or ribosomal target modification (macrolide-lincosamide-streptogramin B [MLSB] resistance; usually encoded by ermA or ermC). MLSB resistance is either constitutive or inducible following exposure to a macrolide. Induction tests utilize closely approximated erythromycin and clindamycin disks; the flattening of the clindamycin zone adjacent to the erythromycin disk indicates inducible MLSB resistance. The present study reassessed the reliability of placing erythromycin and clindamycin disks in adjacent positions (26 to 28 mm apart) in a standard disk dispenser, compared to distances of 15 or 20 mm. A group of 130 clinical isolates of Staphylococcus aureus and 100 isolates of erythromycin resistant coagulase-negative staphylococci (CNS) were examined by disk approximation; all CNS isolates and a subset of S. aureus isolates were examined by PCR for ermA, ermC, and msrA. Of 114 erythromycin-resistant S. aureus isolates, 39 demonstrated constitutive resistance to clindamycin, while 33 showed inducible resistance by disk approximation at all three distances. Only one isolate failed to clearly demonstrate induction at 26 mm. Of 82 erythromycin resistant CNS isolates that contained ermA or ermC, 57 demonstrated constitutive clindamycin resistance, and 25 demonstrated inducible resistance, at 20 and 26 mm. None of the 42 S. aureus isolates or 18 CNS isolates containing only msrA and none of the erythromycin-susceptible isolates yielded positive disk approximation tests. Simple placement of erythromycin and clindamycin disks at a distance achieved with a standard disk dispenser allowed detection of 97% of S. aureus strains and 100% of CNS strains with inducible MLSB resistance in this study. PMID- 14532214 TI - Longitudinal study of intestinal Entamoeba histolytica infections in asymptomatic adult carriers. AB - To gain insight into the dynamics of intestinal Entamoeba histolytica infection, a longitudinal study was performed over an observation period of 15 months with a group of 383 randomly selected adult individuals (mean age, 38.5 years) living in an area of amebiasis endemicity in central Vietnam. Ameba infection was diagnosed by using species-specific PCR and DNA extracted directly from fecal samples. The results indicated an E. histolytica prevalence of 11.2% and an annual new infection rate of 4.1% in the study population. Follow-up of the 43 individuals who were E. histolytica positive at enrollment suggested a regular exponential decline in infection of about 3% per month and a mean half-life of infection of more than 15 months. However, the reinfection rate for this group of participants was 2.7 times higher than that predicted for the study population as a whole. Both the reappearance of the parasite after successful treatment of E. histolytica infection and changes in "genetic fingerprints" of parasites during the course of infection revealed an annual new infection rate of about 11.5%. Thus, the mean half-life of E. histolytica infection was calculated to be 12.9 months (95% confidence interval, 10.2 to 15.6 months). Notably, none of the participants developed symptoms compatible with invasive intestinal amebiasis, and only one of the subjects developed an amebic liver abscess during the observation period. PMID- 14532215 TI - Direct susceptibility testing of positive blood cultures by using Sensititre broth microdilution plates. AB - Traditional susceptibility testing of blood cultures requires overnight incubation in order to obtain isolated colonies. Susceptibility results can be reported up to 24 h sooner by using a bacterial pellet from the blood culture broth. This study evaluated the accuracy of direct susceptibility testing from positive ESP blood culture broths by using Sensititre broth microdilution plates compared to testing with isolated colonies. Practical inclusion criteria were applied to gram-positive organisms to avoid reporting susceptibilities for probable contaminants. All gram-negative organisms were tested directly. An aliquot of the blood culture was centrifuged, and the resulting pellet was used to make a 0.5 McFarland suspension. Microdilution plates were inoculated and interpreted according to the manufacturer's instructions. Colony counts were performed to ensure proper colony density was achieved. A total of 199 patient and seeded blood cultures were evaluated for both essential (within +/-1 twofold dilution) and categorical (sensitive, intermediate, or resistant) agreement. Testing of 93 gram-positive isolates (1,214 antimicrobial agent-organism combinations) yielded 98% essential agreement and categorical error rates of 0.3% minor, no major (false resistance), and 1.7% very major (false susceptibility) errors. For 106 gram-negative isolates (1,828 antimicrobial agent-organism combinations), the essential agreement was 99%. Categorical error rates were 0.5, 0, and 2.0% for minor, major, and very major errors, respectively. Performance was comparable for both gram-positive and gram-negative isolates, as well as for both aerobic and anaerobic media. Using this direct testing methodology, reliable susceptibility results can be reported to physicians 24 h sooner, allowing earlier appropriate modification of antimicrobial therapy. PMID- 14532216 TI - Microwave sterilization of femoral head allograft. AB - The potential shortage of allograft bone has led to the need to investigate other sources of bone for allografts. Some allograft bone donated from primary total hip arthroplasty recipients must be discarded or treated to become usable as a result of bacterial contamination. Femoral head allografts were contaminated with Staphylococcus aureus and Bacillus subtilis. A domestic microwave oven was used. The contaminated bone was exposed to microwave irradiation for different time periods. The samples were then cultured to attempt to grow the two bacterial species. The contaminated bone samples failed to grow any organisms after 2 min of exposure to microwave irradiation. This study shows that sterilization of femoral head allografts contaminated with S. aureus and B. subtilis can be achieved with microwave irradiation in a domestic microwave oven. This method of sterilization of bone allografts is cheap, easily used, and an effective way to process contaminated bone. PMID- 14532217 TI - Use of denaturing high-performance liquid chromatography to identify Bacillus anthracis by analysis of the 16S-23S rRNA interspacer region and gyrA gene. AB - Denaturing high-performance liquid chromatography (DHPLC) was evaluated as a method for identifying Bacillus anthracis by analyzing two chromosomal targets, the 16S-23S intergenic spacer region (ISR) and the gyrA gene. The 16S-23S ISR was analyzed by this method with 42 strains of B. anthracis, 36 strains of Bacillus cereus, and 12 strains of Bacillus thuringiensis; the gyrA gene was analyzed by this method with 33 strains of B. anthracis, 27 strains of B. cereus, and 9 strains of B. thuringiensis. Two blind panels of 45 samples each were analyzed to evaluate the potential diagnostic capability of this method. Our results show that DHPLC is an efficient method for the identification of B. anthracis. PMID- 14532218 TI - Species diversity and polymorphism in the Exophiala spinifera clade containing opportunistic black yeast-like fungi. AB - A monophyletic group of black yeast-like fungi containing opportunistic pathogens around Exophiala spinifera is analyzed using sequences of the small-subunit (SSU) and internal transcribed spacer (ITS) domains of ribosomal DNA. The group contains yeast-like and annellidic species (anamorph genus Exophiala) in addition to sympodial taxa (anamorph genera Ramichloridium and Rhinocladiella). The new species Exophiala oligosperma, Ramichloridium basitonum, and Rhinocladiella similis are introduced and compared with their morphologically similar counterparts at larger phylogenetic distances outside the E. spinifera clade. Exophiala jeanselmei is redefined. New combinations are proposed in EXOPHIALA: Exophiala exophialae for Phaeococcomyces exophialae and Exophiala heteromorpha for E. jeanselmei var. heteromorpha. PMID- 14532219 TI - Exophiala oligosperma causing olecranon bursitis. AB - A 62-year-old male with a history of Wegener's granulomatosis and immunosuppressive therapy presented with chronic olecranon bursitis. A black velvety mould with brown septate hyphae and tapered annellides was isolated from a left elbow bursa aspirate and was identified as an Exophiala species. Internal transcribed sequence rRNA sequencing showed the isolate to be identical to Exophiala oligosperma. The patient was successfully treated with aspiration and intrabursal amphotericin B. PMID- 14532220 TI - Rapid and specific detection of Mycobacterium tuberculosis by using the Smart Cycler instrument and a specific fluorogenic probe. AB - A procedure using the Smart Cycler instrument and a fluorescence quencher (FQ) probe for the specific identification of Mycobacterium tuberculosis complex (MTB) was used to detect organisms in 366 acid-fast bacillus smear-positive respiratory specimens. It was compared to culture and the AMPLICOR M. tuberculosis PCR test. MTB was isolated from 198 of these samples. The FQ PCR assay was sensitive (197 of 198, 99.5%) and specific (165 of 168, 98.2%); no significant difference was observed between the two PCR protocols. After DNA extraction, a final result was available within 1.5 h with the real-time PCR protocol. PMID- 14532221 TI - Differentiation of Candida dubliniensis from Candida albicans on Pal's agar. AB - Production of a hyphal fringe around colonies grown on Pal's agar (sunflower seed agar) at 30 degrees C for 48 to 72 h provides a simple means of discriminating between isolates of C. dubliniensis and C. albicans with 100% accuracy. Of 128 C. dubliniensis isolates tested on this medium, all produced a hyphal fringe. In contrast, none of the 124 C. albicans isolates tested produced a hyphal fringe. Pal's medium has the added advantage of being prepared from inexpensive, readily available seeds. PMID- 14532222 TI - Assessment of immunoglobulin M enzyme immunoassays for diagnosis of measles. AB - We evaluated the performance of three commercial measles immunoglobulin M enzyme immunoassays from Meddens, Denka Seiken, and Behring. The sensitivities were determined to be 96.7% for the Meddens and Denka Seiken assays and 87.9% for the Behring assay. The specificities of the assays were determined to be 94.6% for Meddens, 98.2% for Denka Seiken, and 98.7% for Behring. PMID- 14532223 TI - Use of hepatitis B virus DNA quantitation to predict hepatitis B e antigen reversion in cases of chronic hepatitis B. AB - Seventy-three chronic hepatitis B patients who had either hepatitis B e antigen (HBeAg) seroconversion (group I) or HBeAg-negative disease (group II) were studied. HBV DNA levels at HBeAg seroconversion (group I) and at initial visits (group II) were significantly lower among patients who were persistently negative for HBeAg than among those who underwent HBeAg reversion. PMID- 14532224 TI - Optimization of real-time PCR assay for rapid and sensitive detection of eubacterial 16S ribosomal DNA in platelet concentrates. AB - A real-time PCR assay was developed for rapid detection of eubacterial 16S ribosomal DNA in platelet concentrates. The sensitivity of this assay can be hampered by contaminating DNA in the PCR reagents. Digestion of the PCR reagents with Sau3AI prior to PCR amplification was effective in eliminating this contaminating DNA without affecting the sensitivity of the assay. PMID- 14532225 TI - Genetic diversity among clinical isolates of Candida glabrata analyzed by randomly amplified polymorphic DNA and multilocus enzyme electrophoresis analyses. AB - The genetic diversity of 47 clinical and reference strains of Candida glabrata from several geographical origins and diverse clinical disorders, with different antifungal susceptibilities, as well as their genetic relationships were studied through multilocus enzyme electrophoresis (MLEE) and randomly amplified polymorphic DNA (RAPD) techniques. The genetic diversity estimated for 11 MLEE loci measured as average heterozygosity (h) was 0.055. A high level of genetic relatedness among isolates was established by cluster analysis. Forty-nine RAPD markers were analyzed, and the average genetic diversity among isolates, estimated by Shannon's index (Ho), was 0.372. The PhiST values estimated through an analysis of molecular variance to assess genetic differentiation among isolates revealed no genetic differentiation among them. Our results revealed very low genetic diversity among isolates, a lack of differentiation, and no association with their geographic origin and the clinical characteristics. PMID- 14532226 TI - Comparison of PCR assay to culture for surveillance detection of vancomycin resistant enterococci. AB - Direct multiplex PCR assay using vanA and vanB primers, which provides rapid results, was more sensitive than culture on selective media for samples collected by rectal swab (20 of 46 versus 8 of 46; P < 0.001) or perianal swab (17 of 58 versus 12 of 58; P = 0.059) for the detection of gastrointestinal colonization by vancomycin-resistant enterococci. PMID- 14532227 TI - National Department of Defense surveillance data for antibiotic resistance and emm gene types of clinical group A streptococcal isolates from eight basic training military sites. AB - Antibiotic resistance and emm gene types were examined from 692 Group A streptococci isolates from eight United States military basic training sites between 1998 and 2001. Macrolide resistance was associated with geographic sites and emm type. These data are useful for vaccine development initiatives and antimicrobial treatment considerations. PMID- 14532228 TI - Cellular fatty acid profile distinguishes Burkholderia pseudomallei from avirulent Burkholderia thailandensis. AB - Burkholderia pseudomallei, the cause of melioidosis, can be distinguished from the closely related but nonpathogenic Burkholderia thailandensis by gas chromatography (GC) analysis of fatty acid derivatives. A 2-hydroxymyristic acid derivative (14:0 2OH) was present in 95% of B. pseudomallei isolates and no B. thailandensis isolates. GC mass spectrophotometry confirmed that 2 hydroxymyristic acid was present in B. pseudomallei. GC-fatty acid methyl ester analysis may be useful in distinguishing these two closely related species. PMID- 14532229 TI - Real-time PCR assay targets the 23S-5S spacer for direct detection and differentiation of Legionella spp. and Legionella pneumophila. AB - A real-time PCR for the ABI Prism 7000 system targeting the 23S-5S spacer of Legionella spp. was developed. Simultaneous detection and differentiation of Legionella spp. and Legionella pneumophila within 90 min and without post-PCR melting-curve analysis was achieved using two TaqMan probes. In sputum samples from 23 controls and 17 patients with legionellosis, defined by positive culture, urinary antigen testing, or seroconversion, 94% sensitivity and 100% specificity were observed. PMID- 14532230 TI - Comparison of in vitro activities of 17 antifungal drugs against a panel of 20 dermatophytes by using a microdilution assay. AB - The in vitro activities of 17 antifungal drugs against a panel of 20 dermatophytes comprising 6 different species were determined using a microdilution assay according to the NCCLS M38-P method with some modifications. Terbinafine was the most potent systemic drug while tolnaftate and amorolfine were the most active topical agents. PMID- 14532231 TI - Amplified-fragment length polymorphism as a complement to IS6110-based restriction fragment length polymorphism analysis for molecular typing of Mycobacterium tuberculosis. AB - The amplified-fragment length polymorphism (AFLP) technique was applied to clusters of Mycobacterium tuberculosis clinical isolates obtained by using IS6110 based restriction fragment length polymorphism (RFLP). Ten of the RFLP clusters showed identical AFLP patterns also, but the other 13 could be resolved into subclusters by AFLP. Our results suggest that some RFLP clusters may not be due to recent transmission and that AFLP may be a useful complementary technique. PMID- 14532232 TI - Investigation of Staphylococcus aureus isolates identified as erythromycin intermediate by the Vitek-1 System: comparison with results obtained with the Vitek-2 and Phoenix systems. AB - We identified 69 Staphylococcus aureus isolates that were erythromycin intermediate as reported by the Vitek-1 system using the GPS-105 card. Of the 57 strains that were available for further testing, all were erythromycin resistant by broth microdilution and the Phoenix system, while the Vitek-2 system identified 55 of 57 strains (96%) as erythromycin resistant. The majority of isolates (54 of 57 [95%]) exhibited the inducible MLS (macrolide-lincosamide streptogramin family) phenotype, as shown by the double-disk test. We recommend that all S. aureus strains determined as erythromycin intermediate by the Vitek-1 system be interpreted as resistant to erythromycin. PMID- 14532233 TI - Diagnosis of congenital toxoplasmosis: comparison of targets for detection of Toxoplasma gondii by PCR. AB - Three PCR targets (18S ribosomal DNA, B1, and AF146527) and mouse inoculation were compared for 83 samples in the context of congenital toxoplasmosis. These four techniques are not statistically different in terms of sensitivity and specificity. However, further analysis highlighted problems sometimes encountered with PCR diagnosis of congenital toxoplasmosis. PMID- 14532234 TI - Improved PCR for detection of the highly leukotoxic JP2 clone of Actinobacillus actinomycetemcomitans in subgingival plaque samples. AB - The JP2 clone of Actinobacillus actinomycetemcomitans is associated with early onset periodontitis in certain ethnic populations of African origin. Here, we describe and evaluate a set of primers for PCR to assay for the presence of A. actinomycetemcomitans and to discriminate between JP2-like strains and other genotypes in subgingival plaque samples. PMID- 14532235 TI - Subtyping of Salmonella enterica serovar typhimurium outbreak strains isolated from humans and animals in Iceland. AB - A total of 75 Salmonella enterica serovar Typhimurium strains of various (mainly human and animal) origins were typed by pulsed-field gel electrophoresis (PFGE) and phage typing. These strains were collected during an outbreak in Iceland in 1999 and 2000. The typing revealed that 84% of the strains belonged to the same PFGE and phage type (PT), namely, PFGE type 1Aa and PT 1. PMID- 14532236 TI - Dissemination of extended-spectrum beta-lactamase-producing Enterobacteriaceae in pediatric intensive care units. AB - To study the growing trend of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in pediatric intensive care units (PICUs), 88 nonrepetitive ESBL-producing isolates were prospectively collected and analyzed by molecular methods during a 16-month period. The emergence and dissemination of ESBL producing Enterobacteriaceae in PICUs are the consequence of the clonal dissemination of a few epidemic strains along with the horizontal transmission of resistance gene-carrying plasmids among bacterial organisms. PMID- 14532237 TI - Use of cefoperazone MacConkey agar for selective isolation of Laribacter hongkongensis. AB - A new selective medium, cefoperazone MacConkey agar (CMA), was developed for primary isolation of Laribacter hongkongensis from stool. Its performance in quantitative recovery and in a clinical evaluation of 4,741 human diarrheal stool specimens was superior to that of charcoal cefoperazone deoxycholate agar. In addition, with CMA, Arcobacter butzleri was unexpectedly isolated from the stools of six patients. PMID- 14532238 TI - Volume effect on sensitivity of nucleic acid amplification tests for detection of Chlamydia trachomatis in urine specimens from females. AB - Nucleic acid amplification tests (NAATs) for the detection of Chlamydia trachomatis are routinely used on first-catch urine (FCU) specimens. We analyzed data from a head-to-head comparison of NAATs on female FCU specimens and found that the volume of urine collected could affect test performance. PMID- 14532239 TI - Differentiation of Mycoplasma species by 16S ribosomal DNA PCR and denaturing gradient gel electrophoresis fingerprinting. AB - Denaturing gradient gel electrophoresis (DGGE) of a 16S ribosomal DNA PCR product was used to differentiate 32 mycoplasma species of veterinary significance. Twenty-seven (85%) species could be differentiated by DGGE. This method could enable the rapid identification of many mycoplasma species for which there is no specific PCR available and which are currently identified by using culture and serological tests. PMID- 14532240 TI - Detection of differences in the nucleotide and amino acid sequences of diphtheria toxin from Corynebacterium diphtheriae and Corynebacterium ulcerans causing extrapharyngeal infections. AB - While Corynebacterium ulcerans can mimic classical diphtheria, extrapharyngeal infections are extremely rare. Sequencing of the diphtheria toxin (DT)-encoding tox gene of two C. ulcerans isolates from extrapharyngeal infections revealed differences from C. diphtheriae DT sequences, mainly in the translocation and receptor-binding domains. C. ulcerans supernatants were much less potent than supernatant from C. diphtheriae. A C. ulcerans DT-specific PCR is described below. PMID- 14532241 TI - Biochemical properties of a newly described Escherichia species, Escherichia albertii. AB - Five strains of a newly described Escherichia species, Escherichia albertii, were extensively characterized by conventional biochemical methods and by commercial identification panels. E. albertii is an indole-negative species that ferments D mannitol but not D-xylose. Because these strains are not included in the databases of commercial systems at present, they were most often identified as Hafnia, Salmonella, Escherichia coli, or, on one system (MicroScan dried overnight panels), Yersinia ruckeri. PMID- 14532242 TI - Evaluation of the TRUGENE HCV 5'NC genotyping kit with the new GeneLibrarian module 3.1.2 for genotyping of hepatitis C virus from clinical specimens. AB - The TRUGENE HCV 5'NC genotyping kit (GeneLibrarian modules 3.1.1 and 3.1.2) and VERSANT HCV genotyping assay were compared by using 96 hepatitis C virus (HCV) RNA-positive patient specimens, including HCV genotypes 1, 2, 3, 4, 5, 6, and 10. The TRUGENE HCV 5'NC genotyping kit (GeneLibrarian module 3.1.2) yielded the most accurate genotyping results. PMID- 14532243 TI - Molecular analysis of group A streptococcal isolates associated with scarlet fever in southern Taiwan between 1993 and 2002. AB - Collected between 1993 and 2002 at a Taiwanese university hospital, 77 group A streptococcus isolates associated with scarlet fever were grouped by emm typing and pulsed-field gel electrophoresis. The predominance of an emm1 clone before 1996 and the presence of genetically diverse emm1 and emm4 strains thereafter were found. PMID- 14532244 TI - High disease burden of diarrhea due to enterotoxigenic Escherichia coli among rural Egyptian infants and young children. AB - The incidence of enterotoxigenic Escherichia coli diarrhea among Egyptian children was 1.5 episodes per child per year and accounted for 66% of all first episodes of diarrhea after birth. The incidence increased from 1.7 episodes per child per year in the first 6 months of life to 2.3 in the second 6 months and declined thereafter. PMID- 14532245 TI - Photographic and luminometric detection of luciferase reporter phages for drug susceptibility testing of clinical Mycobacterium tuberculosis isolates. AB - Luciferase reporter phages (LRPs) have proven to be efficient tools for drug susceptibility testing of Mycobacterium tuberculosis. Luminometric detection of LRP activity offers higher sensitivity and quantitative results, while a Polaroid film detection method offers a "low-tech" inexpensive alternative that is called the Bronx box. In this work we evaluated, improved, and compared the performance of the luminometer and the Bronx box formats for drug susceptibility testing with LRPs by using 51 clinical isolates of M. tuberculosis, with the agar proportion method (PM) serving as reference. The sensitivity in detecting resistance to isoniazid and rifampin, antibiotics that define multidrug resistance (MDR), was 100% for both methods. The turnaround time for results was reduced from 3 weeks for PM to 54 or 94 h for luminometry or the Bronx box, respectively. These results support the utility of LRPs as a screening test for the surveillance of MDR tuberculosis. PMID- 14532246 TI - Temporal and geographical distribution and overlap of Penner heat-stable serotypes and pulsed-field gel electrophoresis genotypes of Campylobacter jejuni isolates collected from humans and chickens in Finland during a seasonal peak. AB - The association of Penner heat-stable serotypes and pulsed-field gel electrophoresis genotypes of 208 human and 30 chicken Campylobacter jejuni isolates was studied. Overall, 46% of the human strains had overlapping sero- and genotype combinations with chicken strains. The percentage was reduced to 31% for strains that were considered temporally related. This suggests common environmental sources. PMID- 14532247 TI - Detection of Yersinia pestis in sputum by real-time PCR. AB - A 5' nuclease PCR assay for detection of the Yersinia pestis plasminogen activator (pla) gene in human respiratory specimens with simulated Y. pestis infection was developed. An internal positive control was added to the reaction mixture in order to detect the presence of PCR inhibitors that are often found in biological samples. The assay was 100% specific for Y. pestis. In the absence of inhibitors, a sensitivity of 10(2) CFU/ml of respiratory fluid was obtained. When inhibitors were present, detection of Y. pestis DNA required a longer sample treatment time and an initial concentration of bacteria of at least 10(4) CFU/ml. The test's total turnaround time was less than 5 h. The assay described here is well suited to the rapid diagnosis of pneumonic plague, the form of plague most likely to result from a bioterrorist attack. PMID- 14532248 TI - Epidemiological study of resistance to nalidixic acid and other antibiotics in clinical Yersinia enterocolitica O:3 isolates. AB - Forty-six Yersinia enterocolitica O:3 clinical isolates resistant to nalidixic acid were studied. The use of molecular typing techniques, other indicators of resistance patterns, the plasmid profile, and the presence of genes that encode aminoglycoside-modifying enzyme production suggested to us a clonal dissemination of the studied strains. PMID- 14532249 TI - Surveillance for respiratory syncytial virus in infants hospitalized for acute lower respiratory infection in Chile (1989 to 2000). AB - Hospitalized infants (4,618) were studied for lower respiratory infections from 1989 through 2000 by routine immunofluorescence assay and viral isolation. The hospitalization rate for respiratory syncytial virus (RSV) averaged 2% per year. The fatality rate was 0.1%. Monthly RSV detection varied from 14 to 88%, and epidemics lasted 3.5 to 6 months. From 1994 high-early versus low-late epidemic patterns alternately were observed, the first influenced by a group B strain. PMID- 14532250 TI - Staphylococcal exanthematous disease in a newborn due to a virulent methicillin resistant Staphylococcus aureus strain containing the TSST-1 gene in Europe: an alert for neonatologists. AB - We report a case of staphylococcal exanthematous disease in a newborn due to a toxic shock syndrome toxin 1- and SEC-producing methicillin-resistant Staphylococcus aureus strain and alert neonatologists to the probable emergence in France of the neonatal toxic shock syndrome-like exanthematous disease in newborns previously described in Japan. We advise neonatologists to pay careful attention to clinical parameters and to prescribe appropriate tests: platelet count, serum C-reactive protein concentration, and Vbeta2-positive T-cell counts. PMID- 14532251 TI - Bipolaris spicifera causes fungus balls of the sinuses and triggers polypoid chronic rhinosinusitis in an immunocompetent patient. AB - We report the rare case of a 19-year-old immunocompetent male suffering both from fungus balls of the sinuses and from chronic rhinosinusitis with massive polyposis. Endoscopic sinus surgery revealed grayish brown necrotic masses embedded in viscous eosinophilic mucus. Inoculated onto petri dishes, these masses as well as the mucus grew a dark pigmented fungus, which was identified as Bipolaris spicifera. PMID- 14532252 TI - Molecular characterization of a strain of group a streptococcus isolated from a patient with a psoas abscess. AB - We report the first case of a primary group A streptococcus (GAS) psoas abscess in a 31-year-old woman. The psoas abscess was preceded by an episode of acute pharyngitis. The M-protein gene (emm) and streptolysin S structural gene (sagA) were present in the isolate, with no significant amino acid differences from previously described sequences of M1 GAS isolates. Multilocus sequence typing (MLST) showed that the isolate belonged to MLST sequence type (MLST-ST) 28, the predominant MLST-ST associated with invasive disease caused by M1 isolates. PMID- 14532253 TI - Serial granulocyte transfusions as a treatment for sepsis due to multidrug resistant Pseudomonas aeruginosa in a neutropenic patient. AB - The emergence of multidrug-resistant Pseudomonas aeruginosa (MRPA) has become a major clinical problem. We successfully treated MRPA sepsis in a neutropenic patient undergoing peripheral blood stem cell transplantation with serial granulocyte transfusions. Granulocyte transfusion should be considered as a treatment for severe infection in patients with neutropenia. PMID- 14532254 TI - Occurrence of renal and pulmonary syndrome in a region of northeast Germany where Tula hantavirus circulates. AB - Hantavirus species Tula (TULV) is carried by European common voles (Microtus spp.). Its pathogenic potential for humans is unknown. In a rural region of northeast Germany, a 43-year-old man became ill with fever, renal syndrome, and pneumonia. Typing of late acute- and convalescent-phase sera by focus reduction neutralization assay revealed the presence of neutralizing antibodies against TULV. Moreover, we detected TULV genetic material in Microtus arvalis animals that were trapped at places only a few kilometers from the home village of the patient. Phylogenetic analysis of completely sequenced genomic S segments from three virus strains grouped them within a third genetic lineage of the TULV species. This is the first case of hemorrhagic fever with renal syndrome and pulmonary involvement which can be associated with TULV infection. PMID- 14532255 TI - Successful outcome of disseminated Fusarium infection with skin localization treated with voriconazole and amphotericin B-lipid complex in a patient with acute leukemia. AB - A disseminated Fusarium oxysporum infection with skin localization was diagnosed in a woman with a relapse of B-acute leukemia during induction chemotherapy. The infection was refractory to amphotericin B-lipid complex alone but responded successfully when voriconazole was added. PMID- 14532256 TI - Paronychia due to Prevotella bivia that resulted in amputation: fast and correct bacteriological diagnosis is crucial. AB - Prevotella bivia is mainly associated with endometritis. The case of a patient with paronychia in a thumb due to P. bivia resulting in osteitis and amputation is reported. The species was not acknowledged in the first bacterial culture 2 weeks before surgery. PMID- 14532257 TI - Persistent multiple pulmonary nodules in a nonimmunocompromised woman after varicella-related myelitis treated with acyclovir. AB - Persistent multiple pulmonary nodules were observed on the chest X ray of a nonimmunocompromised woman 6 months after she was treated with acyclovir for a varicella-related myelitis without respiratory symptoms. Early antiviral therapy given for varicella infections might decrease the intensity of clinical symptoms without actually preventing the occurrence of varicella-zoster virus-related lesions such as the persistent pulmonary nodules reported here. PMID- 14532258 TI - Macrolide resistance in Streptococcus pneumoniae strains collected in the Far East of Russia from 2000 to 2002. PMID- 14532259 TI - Heterogeneity of Mycobacterium tuberculosis isolates in Yangon, Myanmar. PMID- 14532260 TI - Molecular characterization of anthrax in positive powders: a Mexican experience. PMID- 14532261 TI - Sterilization of skin and catheters before drawing blood cultures. PMID- 14532262 TI - Need for procedural details in the protocol for specimen processing by the MagNA Pure LC Instrument. PMID- 14532263 TI - Sequence-specific dimerization of the transmembrane domain of the "BH3-only" protein BNIP3 in membranes and detergent. AB - Mitochondria-mediated apoptosis is regulated by proteins of the Bcl-2 superfamily, most of which contain a C-terminal hydrophobic domain that plays a role in membrane targeting. Experiments with BNIP3 have implicated the transmembrane (TM) domain in its proapoptotic function, homodimerization, and interactions with Bcl-2 and Bcl-xL. We show that the BNIP3 TM domain self associates strongly in Escherichia coli cell membranes and causes reversible dimerization of a soluble protein in the detergent SDS when expressed as an in frame fusion. Limited mutational analysis identifies specific residues that are critical for BNIP3 TM self-association in membranes, and these residues are also important for dimerization in SDS micelles, suggesting that the self-association observed in membranes is preserved in detergent. The effects of sequence changes at positions Ala176 and Gly180 suggest that the BNIP3 TM domain associates using a variant of the GXXXG motif previously shown to be important in the dimerization of glycophorin A. The importance of residue His173 in BNIP3 TM domain dimerization indicates that polar residues, which have been implicated in self association of model TM peptides, can act in concert with the AXXXG motif to stabilize TM domain interactions. Our results demonstrate that the hydrophobic C terminal TM domain of the pro-apoptotic BNIP3 protein dimerizes tightly in lipidic environments, and that this association has a strong sequence dependence but is independent of the identity of flanking regions. Thus, the transmembrane domain represents another region of the Bcl-2 superfamily of proteins that is capable of mediating strong and specific protein-protein interactions. PMID- 14532264 TI - Silencing of Sm proteins in Trypanosoma brucei by RNA interference captured a novel cytoplasmic intermediate in spliced leader RNA biogenesis. AB - In Trypanosoma brucei the small nuclear (sn) RNAs U1, U2, U4, and U5, as well as the spliced leader (SL) RNA, bind the seven Sm canonical proteins carrying the consensus Sm motif. To determine the function of these proteins in snRNA and SL RNA biogenesis, two of the Sm core proteins, SmE and SmD1, were silenced by RNAi. Surprisingly, whereas the level of all snRNAs, including U1, U2, U4, and U5 was reduced during silencing, the level of SL RNA was dramatically elevated, but the levels of U6 and spliced leader-associated RNA (SLA1) remained unchanged. The SL RNA that had accumulated in silenced cells lacked modification at the cap4 nucleotide but harbored modifications at the cap1 and cap2 nucleotides and carried the characteristic psi. This SL RNA possessed a longer tail and had accumulated in the cytoplasm in 10 and 50 S particles that were found by in situ hybridization to be present in "speckles." We propose a model for SL RNA biogenesis involving a cytoplasmic phase and suggest that the trypanosome specific "cap4" nucleotides function as a signal for export and import of SL RNA out and into the nucleus. The SL RNA biogenesis pathway differs from that of U sn ribonucleoproteins (RNPs) in that it is the only RNA that binds Sm proteins that were stabilized under Sm depletion in a novel RNP, which we termed SL RNP-C. PMID- 14532265 TI - Organic solutes rescue the functional defect in delta F508 cystic fibrosis transmembrane conductance regulator. AB - The most common defect in cystic fibrosis, deletion of phenylalanine from position 508 of the cystic fibrosis transmembrane conductance regulator (Delta F508 CFTR), decreases the trafficking of this protein to the cell surface membrane. Previous studies have shown that low temperature and high concentrations of glycerol or trimethylamine N-oxide can partially counteract the processing defect of Delta F508 CFTR. The present study investigates whether physiologically relevant concentrations of organic solutes, accumulated by cotransporter proteins, can rescue the misprocessing of Delta F508 CFTR. Myoinositol alone or myoinositol, betaine, and taurine given sequentially increased the processing of core-glycosylated, endoplasmic reticulum-arrested Delta F508 CFTR into the fully glycosylated form of CFTR in IB3 cells or NIH 3T3 cells stably expressing Delta F508 CFTR. Pulse-chase experiments using transiently transfected COS7 cells demonstrated that organic solutes also increased the processing of the core-glycosylated form of green fluorescent protein-Delta F508 CFTR. Moreover, the prolonged half-life of the complex glycosylated form of GFP-Delta F508 CFTR suggests that this treatment stabilized the mature form of the protein. In vitro studies of purified NBD1 stability and aggregation showed that myoinositol stabilized both the Delta F508 and wild type CFTR and inhibited Delta F508 misfolding. Most significantly, treatment of CF bronchial airway cells with these transportable organic solutes restores cAMP stimulated single channel activity of both CFTR and outwardly rectifying chloride channel in the cell surface membrane and also restores a forskolin-stimulated macroscopic 36Cl- efflux. We conclude that organic solutes can repair CFTR functions by enhancing the processing of Delta F508 CFTR to the plasma membrane by stabilizing the complex-glycosylated form of Delta F508 CFTR. PMID- 14532266 TI - Mutagenesis studies of protein farnesyltransferase implicate aspartate beta 352 as a magnesium ligand. AB - Protein farnesyltransferase (FTase) catalyzes the addition of a farnesyl chain onto the sulfur of a C-terminal cysteine of a protein substrate. Magnesium ions enhance farnesylation catalyzed by FTase by several hundred-fold, with a KMg value of 4 mM. The magnesium ion is proposed to coordinate the diphosphate leaving group of farnesyldiphosphate (FPP) to stabilize the developing charge in the farnesylation transition state. Here we further investigate the magnesium binding site using mutagenesis and biochemical studies. Free FPP binds Mg2+ with a Kd of 120 microM. The 10-fold weaker affinity for Mg2+ observed for the FTase.FPP.peptide ternary complex is probably caused by the positive charges in the diphosphate binding pocket of FTase. Furthermore, mutation of aspartate beta 352 to alanine (D beta 352A) or lysine (D beta 352K) in FTase drastically alters the Mg2+ dependence of FTase catalysis without dramatically affecting the rate constant of farnesylation minus magnesium or the binding affinity of either substrate. In D beta 352A FTase, the KMg increases 28-fold to 110 +/- 30 mM, and the farnesylation rate constant at saturating Mg2+ decreases 27-fold to 0.30 +/- 0.05 s-1. Substitution of a lysine for Asp-beta 352 removes the magnesium activation of farnesylation catalyzed by FTase but does not significantly enhance the rate constant for farnesylation in the absence of Mg2+. In wild type FTase, Mg2+ can be replaced by Mn2+ with a 2-fold lower KMn (2 mM). These results suggest both that Mg2+ coordinates the side chain carboxylate of Asp-beta 352 and that the role of magnesium in the reaction includes positioning the FPP prior to catalysis. PMID- 14532267 TI - Localization of an antithrombin exosite that promotes rapid inhibition of factors Xa and IXa dependent on heparin activation of the serpin. AB - We have previously shown that exosites in antithrombin outside the P6-P3' reactive loop region become available upon heparin activation to promote rapid inhibition of the target proteases, factor Xa and factor IXa. To identify these exosites, we prepared six antithrombin-alpha 1-proteinase inhibitor chimeras in which antithrombin residues 224-286 and 310-322 that circumscribe a region surrounding the reactive loop on the inhibitor surface were replaced in 10-16 residue segments with the homologous segments of alpha1-proteinase inhibitor. All chimeras bound heparin with a high affinity similar to wild-type, underwent heparin-induced fluorescence changes indicative of normal conformational activation, and were able to form SDS-stable complexes with thrombin, factor Xa, and factor IXa and inhibit these proteases with stoichiometries minimally altered from those of wild-type antithrombin. With only one exception, conformational activation of the chimeras with a heparin pentasaccharide resulted in normal approximately 100-300-fold enhancements in reactivity with factor Xa and factor IXa. The exception was the chimera in which residues 246-258 were replaced, corresponding to strand 3 of beta-sheet C, which showed little or no enhancement of its reactivity with these proteases following pentasaccharide activation. By contrast, all chimeras including the strand 3C chimera showed essentially wild type reactivities with thrombin after pentasaccharide activation as well as normal full-length heparin enhancements in reactivity with all proteases due to heparin bridging. These findings suggest that antithrombin exosites responsible for enhancing the rates of factor Xa and factor IXa inhibition in the conformationally activated inhibitor lie in strand 3 of beta-sheet C of the serpin. PMID- 14532268 TI - Molecular basis for the direct inhibition of AP-1 DNA binding by 15-deoxy-Delta 12,14-prostaglandin J2. AB - Cyclopentenone prostaglandins may interfere with cellular functions by multiple mechanisms. The cyclopentenone 15-deoxy-Delta 12,14-prostaglandin J2 (15d-PGJ2) has been reported to inhibit the activity of the transcription factor AP-1 in several experimental settings. We have explored the possibility of a direct interaction of 15d-PGJ2 with AP-1 proteins. Here we show that 15d-PGJ2 covalently modifies c-Jun and directly inhibits the DNA binding activity of AP-1. The modification of c-Jun occurs both in vitro and in intact cells as detected by labeling with biotinylated 15d-PGJ2 and mass spectrometry analysis. Attachment of the cyclopentenone prostaglandin occurs at cysteine 269, which is located in the c-Jun DNA binding domain. In addition, 15d-PGJ2 can promote the oligomerization of a fraction of c-Jun through the formation of intermolecular disulfide bonds or 15d-PGJ2-bonded dimers. Our results identify a novel site of interaction of 15d PGJ2 with the AP-1 activation pathway that may contribute to the complex effects of cyclopentenone prostaglandins on the cellular response to pro-inflammatory agents. They also show the first evidence for the induction of protein cross linking by 15d-PGJ2. PMID- 14532269 TI - Mechanisms of growth hormone (GH) action. Identification of conserved Stat5 binding sites that mediate GH-induced insulin-like growth factor-I gene activation. AB - Many of the actions of growth hormone (GH) on somatic growth and tissue maintenance are mediated by insulin-like growth factor-I (IGF-I), a secreted protein whose gene expression is rapidly and potently induced by GH by unknown mechanisms. Recent studies implicating Stat5a and Stat5b in the growth response to GH in mice and observations linking Stat5b to control of IGF-I gene transcription in rats have prompted the current investigations into the molecular determinants of a putative regulatory network extending from GH through Stat5b to IGF-I. Here we characterize as critical components of this hormone-activated transcriptional pathway two adjacent Stat5 binding sites in the second intron of the rat IGF-I gene located within a conserved region previously found to undergo acute and reversible changes in chromatin structure after in vivo GH treatment. As assessed by chromatin immunoprecipitation assays, GH rapidly induced binding of Stat5 to this DNA segment in the liver of GH-deficient rats, just prior to the onset of transcription from both major and minor IGF-I gene promoters. Biochemical reconstitution experiments showed that the two intronic Stat5 DNA elements were able to bind Stat5b in vitro after GH treatment could transmit GH- and Stat5b-dependent transcriptional responsiveness to the major IGF-I promoter and to a minimal neutral gene promoter and were required for full stimulation of reporter gene activity by GH. Taken together, these results identify an intronic enhancer as a key mediator of GH-induced IGF-I gene transcription working through Stat5b and provide new insight into the molecular architecture of this transcriptional pathway. PMID- 14532270 TI - A product of the human gene adjacent to parkin is a component of Lewy bodies and suppresses Pael receptor-induced cell death. AB - Parkin, a RING-type ubiquitin ligase, is the product of the gene responsible for autosomal recessive juvenile parkinsonism. A reverse strand gene located upstream of the parkin gene in the human genome has been identified. The gene product, termed Glup/PACRG, forms a large molecular chaperone complex containing heat shock proteins 70 and 90 and chaperonin components. Glup suppressed cell death induced by accumulation of unfolded Pael receptor (Pael-R), a substrate of Parkin. On the other hand, Glup facilitated the formation of inclusions consisting of Pael-R, molecular chaperones, protein degradation molecules, and Glup itself, when proteasome is inhibited. Glup knockdown attenuated the formation of Pael-R inclusions, which resulted in the promotion of cell death with extensive vacuolization. Moreover, Glup turned out to be a component of Lewy bodies in Parkinson's disease cases. These data suggest that Glup may play an important role in the formation of Lewy bodies and protection of dopaminergic neurons against Parkinson's disease. PMID- 14532271 TI - An unusual peptide deformylase features in the human mitochondrial N-terminal methionine excision pathway. AB - Dedicated machinery for N-terminal methionine excision (NME) was recently identified in plant organelles and shown to be essential in plastids. We report here the existence of mitochondrial NME in mammals, as shown by the identification of cDNAs encoding specific peptide deformylases (PDFs) and new methionine aminopeptidases (MAP1D). We cloned the two full-length human cDNAs and showed that the N-terminal domains of the encoded enzymes were specifically involved in targeting to mitochondria. In contrast to mitochondrial MAP1D, the human PDF sequence differed from that of known PDFs in several key features. We characterized the human PDF fully in vivo and in vitro. Comparison of the processed human enzyme with the plant mitochondrial PDF1A, to which it is phylogenetically related, showed that the human enzyme had an extra N-terminal domain involved in both mitochondrial targeting and enzyme stability. Mammalian PDFs also display non-random substitutions in the conserved motifs important for activity. Human PDF site-directed mutagenesis variants were studied and compared with the corresponding plant PDF1A variants. We found that amino acid substitutions in human PDF specifically altered its catalytic site, resulting in an enzyme intermediate between bacterial PDF1Bs and plant PDF1As. Because (i) human PDF was found to be active both in vitro and in vivo, (ii) the entire machinery is conserved and expressed in most animals, (iii) the mitochondrial genome expresses substrates for these enzymes, and (iv) mRNA synthesis is regulated, we conclude that animal mitochondria have a functional NME machinery that can be regulated. PMID- 14532272 TI - Interactions among gamma R268, gamma Q269, and the beta subunit catch loop of Escherichia coli F1-ATPase are important for catalytic activity. AB - Removal of the ability to form a salt bridge or hydrogen bonds between the beta subunit catch loop (beta Y297-D305) and the gamma subunit of Escherichia coli F1Fo-ATP synthase significantly altered the ability of the enzyme to hydrolyze ATP and the bacteria to grow via oxidative phosphorylation. Residues beta T304, beta D305, beta D302, gamma Q269, and gamma R268 were found to be very important for ATP hydrolysis catalyzed by soluble F1-ATPase, and the latter four residues were also very important for oxidative phosphorylation. The greatest effects on catalytic activity were observed by the substitution of side chains that contribute to the shortest and/or multiple H-bonds as well as the salt bridge. Residue beta D305 would not tolerate substitution with Val or Ser and had extremely low activity as beta D305E, suggesting that this residue is particularly important for synthesis and hydrolysis activity. These results provide evidence that tight winding of the gamma subunit coiled-coil is important to the rate-limiting step in ATP hydrolysis and are consistent with an escapement mechanism for ATP synthesis in which alpha beta gamma intersubunit interactions provide a means to make substrate binding a prerequisite of proton gradient driven gamma subunit rotation. PMID- 14532273 TI - Rpe65 is a retinyl ester binding protein that presents insoluble substrate to the isomerase in retinal pigment epithelial cells. AB - Photon capture by a rhodopsin pigment molecule induces 11-cis to all-trans isomerization of its retinaldehyde chromophore. To restore light sensitivity, the all-trans-retinaldehyde must be chemically re-isomerized by an enzyme pathway called the visual cycle. Rpe65, an abundant protein in retinal pigment epithelial (RPE) cells and a homolog of beta-carotene dioxygenase, appears to play a role in this pathway. Rpe65-/- knockout mice massively accumulate all-trans-retinyl esters but lack 11-cis-retinoids and rhodopsin visual pigment in their retinas. Mutations in the human RPE65 gene cause a severe recessive blinding disease called Leber's congenital amaurosis. The function of Rpe65, however, is unknown. Here we show that Rpe65 specifically binds all-trans-retinyl palmitate but not 11 cis-retinyl palmitate by a spectral-shift assay, by co-elution during gel filtration, and by co-immunoprecipitation. Using a novel fluorescent resonance energy transfer (FRET) binding assay in liposomes, we demonstrate that Rpe65 extracts all-trans-retinyl esters from phospholipid membranes. Assays of isomerase activity reveal that Rpe65 strongly stimulates the enzymatic conversion of all-trans-retinyl palmitate to 11-cis-retinol in microsomes from bovine RPE cells. Moreover, we show that addition of Rpe65 to membranes from rpe65-/- mice, which possess no detectable isomerase activity, restores isomerase activity to wild-type levels. Rpe65 by itself, however, has no intrinsic isomerase activity. These observations suggest that Rpe65 presents retinyl esters as substrate to the isomerase for synthesis of visual chromophore. This proposed function explains the phenotype in mice and humans lacking Rpe65. PMID- 14532274 TI - Dynamic features of a heme delivery system for cytochrome C maturation. AB - In Escherichia coli, heme is delivered to cytochrome c in a process involving eight proteins encoded by the ccmABCDEFGH operon. Heme is transferred to the periplasmic heme chaperone CcmE by CcmC and from there to apocytochrome c. The role of CcmC was investigated by random as well as site-directed mutagenesis. Important amino acids were all located in periplasmic domains of the CcmC protein that has six membrane-spanning helices. Besides the tryptophan-rich motif and two conserved histidines, new residues were identified as functionally important. Mutants G111S and H184Y had a clear defect in CcmC-CcmE interaction, did not transfer heme to CcmE, and lacked c-type cytochromes. Conversely, mutants D47N, R55P, and S176Y were affected neither in interaction with nor in delivery of heme to CcmE but produced less than 10% c-type cytochromes. A strain carrying a CcmCE fusion had a similar phenotype, suggesting that CcmC is important not only for heme transfer to CcmE but also for its delivery to cytochrome c. Co immunoprecipitation of CcmC with CcmF was not detectable although CcmE co precipitated individually with CcmC and CcmF. This contradicts the idea of CcmCEF supercomplex formation. Our results favor a model that predicts CcmE to shuttle between CcmC and CcmF for heme delivery. PMID- 14532275 TI - The intermediate S1' pocket of the endometase/matrilysin-2 active site revealed by enzyme inhibition kinetic studies, protein sequence analyses, and homology modeling. AB - Human matrix metalloproteinase-26 (MMP-26/endometase/matrilysin-2) is a newly identified MMP and its structure has not been reported. The enzyme active site S1' pocket in MMPs is a well defined substrate P1' amino acid residue-binding site with variable depth. To explore MMP-26 active site structure-activity, a series of new potent mercaptosulfide MMP inhibitors (MMPIs) with Leu or homophenylalanine (Homophe) side chains at the P1' site were selected. The Homephe side chain is designed to probe deep S1' pocket MMPs. These inhibitors were tested against MMP-26 and several MMPs with known x-ray crystal structures to distinguish shallow, intermediate, and deep S1' pocket characteristics. MMP-26 has an inhibition profile most similar to those of MMPs with intermediate S1' pockets. Investigations with hydroxamate MMPIs, including those designed for deep pocket MMPs, also indicated the presence of an intermediate pocket. Protein sequence analysis and homology modeling further verified that MMP-26 has an intermediate S1' pocket formed by Leu-204, His-208, and Tyr-230. Moreover, residue 233 may influence the depth of an MMP S1' pocket. The residue at the equivalent position of MMP-26 residue 233 is hydrophilic in intermediate-pocket MMPs (e.g. MMP-2, -8, and -9) and hydrophobic in deep-pocket MMPs (e.g. MMP-3, 12, and -14). MMP-26 contains a His-233 that renders the S1' pocket to an intermediate size. This study suggests that MMPIs, protein sequence analyses, and molecular modeling are useful tools to understand structure-activity relationships and provides new insight for rational inhibitor design that may distinguish MMPs with deep versus intermediate S1' pockets. PMID- 14532276 TI - Characterization of recombinant skeletal muscle (Ser-2843) and cardiac muscle (Ser-2809) ryanodine receptor phosphorylation mutants. AB - Phosphorylation of the skeletal muscle (RyR1) and cardiac muscle (RyR2) ryanodine receptors has been reported to modulate channel activity. Abnormally high phosphorylation levels (hyperphosphorylation) at Ser-2843 in RyR1 and Ser-2809 in RyR2 and dissociation of FK506-binding proteins from the receptors have been implicated as one of the causes of altered calcium homeostasis observed during human heart failure. Using site-directed mutagenesis, we prepared recombinant RyR1 and RyR2 mutant receptors mimicking constitutively phosphorylated and dephosphorylated channels carrying a Ser/Asp (RyR1-S2843D and RyR2-S2809D) and Ser/Ala (RyR1-S2843A and RyR2-S2809A) substitution, respectively. Following transient expression in human embryonic kidney 293 cells, the effects of Ca2+, Mg2+, and ATP on channel function were determined using single channel and [3H]ryanodine binding measurements. In both assays, neither the skeletal nor cardiac mutants showed significant differences compared with wild type. Similarly essentially identical caffeine responses were observed in Ca2+ imaging measurements. Co-immunoprecipitation and Western blot analysis showed comparable binding of FK506-binding proteins to wild type and mutant receptors. Finally metabolic labeling experiments showed that the cardiac ryanodine receptor was phosphorylated at additional sites. Taken together, the results did not support the view that phosphorylation of a single site (RyR1-Ser-2843 and RyR2-Ser-2809) substantially changes RyR1 and RyR2 channel function. PMID- 14532277 TI - Etk/Bmx transactivates vascular endothelial growth factor 2 and recruits phosphatidylinositol 3-kinase to mediate the tumor necrosis factor-induced angiogenic pathway. AB - Tumor necrosis factor (TNF), via its receptor 2 (TNFR2), induces Etk (or Bmx) activation and Etk-dependent endothelial cell (EC) migration and tube formation. Because TNF receptor 2 lacks an intrinsic kinase activity, we examined the kinase(s) mediating TNF-induced Etk activation. TNF induces a coordinated phosphorylation of vascular endothelial growth factor (VEGF) receptor 2 (VEGFR2) and Etk, which is blocked by VEGFR2-specific inhibitors. In response to TNF, Etk and VEGFR2 form a complex resulting in a reciprocal activation between the two kinases. Subsequently, the downstream phosphatidylinositol 3-kinase (PI3K)-Akt signaling (but not signaling through phospholipase C-gamma) was initiated and directly led to TNF-induced EC migration, which was significantly inhibited by VEGFR2-, PI3K-, or Akt-specific inhibitors. Phosphorylation of VEGFR2 at Tyr-801 and Tyr-1175, the critical sites for VEGF-induced PI3K-Akt signaling, was not involved in TNF-mediated Akt activation. However, TNF induces phosphorylation of Etk at Tyr-566, directly mediating the recruitment of the p85 subunit of PI3K. Furthermore, TNF- but not VEGF-induced activation of VEGFR2, Akt, and EC migration are blunted in EC genetically deficient with Etk. Taken together, our data demonstrated that TNF induces transactivation between Etk and VEGFR2, and Etk directly activates PI3K-Akt angiogenic signaling independent of VEGF-induced VEGFR2-PI3K-Akt signaling pathway. PMID- 14532278 TI - Release of arachidonic acid by stimulation of opsonic receptors in human monocytes: the FcgammaR and the complement receptor 3 pathways. AB - The role of the opsonic receptors FcgammaR and CR3 on the release of arachidonic acid (AA) by human monocytes was studied using IgG-ovalbumin (OVA) equivalence immune complexes (IC), anti-OVA IgG bound to OVA-coupled latex beads, and C3bi bound IC. Release of AA was produced by IC and latex-OVA beads bound to IgG, whereas binding of C3bi to IC inhibited the ability of IC to release AA. In contrast, coating of zymosan particles with C3bi enhanced AA release as compared with that produced by non-coated particles. Masking of C3bi on C3bi-bound IC by incubation with anti-C3 IgG resulted in the recovery of their ability to release AA, thereby suggesting that binding of C3b by IC reduces their flogogenic effects, whereas opsonization of microbial walls by complement may enhance their proinflammatory potential. The binding/uptake of opsonized zymosan particles was inhibited by anti-CR3 Ab and C3bi-bound IC, but not by beta-glucan, mannan, and anti-Toll-like receptor 2 Ab. These findings show that cooperative engagement of CR3 on both the lectin-like site involved in beta-glucan binding and the I-domain involved in C3bi binding, as it can be observed in the innate immune response, produces AA release, whereas the unique interaction of C3bi-bound IC with the I domain of CR3, as it may occur in the adaptive immune response, diverts the IC lattice from a productive interaction with FcgammaR linked to AA release. PMID- 14532279 TI - Analysis of transmembrane segment 7 of the dipeptide transporter hPepT1 by cysteine-scanning mutagenesis. AB - To investigate the involvement of transmembrane segment 7 (TMS7) of hPepT1 in forming the putative central aqueous channel through which the substrate traverses, we individually mutated each of the 21 amino acids in TMS7 to a cysteine and analyzed the mutated transporters using the scanning cysteine accessibility method. Y287C- and M292C-hPepT1 did not express at the plasma membrane. Out of the remaining 19 transporters, three (F293C-, L296C-, and F297C hPepT1) showed negligible glycyl-sarcosine (gly-sar) uptake activity and may play an important role in defining the overall hPepT1 structure. K278C-hPepT1 showed approximately 40% activity and the 15 other transporters exhibited more than 50% gly-sar uptake when compared with wild type (WT)-hPepT1. Gly-sar uptake for the 16 active transporters containing cysteine mutations was then measured in the presence of 2.5 mM 2-aminoethyl methanethiosulfonate hydrobromide (MTSEA) or 1 mM [2-(trimethylammonium) ethyl] methanethiosulfonate bromide (MTSET). Gly-sar uptake was significantly inhibited for each of the 16 single cysteine mutants in the presence of 2.5 mM MTSEA. In contrast, significant inhibition of uptake was only observed for K278C-, M279C-, V280C-, T281C-, M284C-, L286C-, P291C-, and D298C-hPepT1 in the presence of 1 mM MTSET. MTSET modification of R282C-hPepT1 resulted in a significant increase in gly-sar uptake. To investigate this further, we mutated WT-hPepT1 to R282A-, R282E-, and R282K-hPepT1. R282E-hPepT1 showed a 43% reduction in uptake activity, whereas R282A- and R282K-hPepT1 had activities comparable with WT-hPepT1, suggesting a role for the Arg-282 positive charge in substrate translocation. Most of the amino acids that were MTSET sensitive upon cysteine mutation, including R282C, are located toward the intracellular end of TMS7. Hence, our results suggest that TMS7 of hPepT1 is relatively solvent-accessible along most of its length but that the intracellular end of the transmembrane domain is particularly so. From a structure-function perspective, we speculate that the extracellular end of TMS7 may shift following substrate binding, providing the basis for channel opening and substrate translocation. PMID- 14532280 TI - Deformation of helix C in the low temperature L-intermediate of bacteriorhodopsin. AB - X-ray and electron diffraction studies of specific reaction intermediates, or reaction intermediate analogues, have produced a consistent picture of the structural mechanism of light-driven proton pumping by bacteriorhodopsin. Of central importance within this picture is the structure of the L-intermediate, which follows the retinal all-trans to 13-cis photoisomerization step of the K intermediate and sets the stage for the primary proton transfer event from the positively charged Schiff base to the negatively charged Asp-85. Here we report the structural changes in bacteriorhodopsin following red light illumination at 150 K. Single crystal microspectrophotometry showed that only the L-intermediate is populated in three-dimensional crystals under these conditions. The experimental difference Fourier electron density map and refined crystallographic structure were consistent with those previously presented (Royant, A., Edman, K., Ursby, T., Pebay-Peyroula, E., Landau, E. M., and Neutze, R. (2000) Nature 406, 645-648; Royant, A., Edman, K., Ursby, T., Pebay-Peyroula, E., Landau, E. M., and Neutze, R. (2001) Photochem. Photobiol. 74, 794-804). Based on the refined crystallographic structures, molecular dynamic simulations were used to examine the influence of the conformational change of the protein that is associated with the K-to-L transition on retinal dynamics. Implications regarding the structural mechanism for proton pumping by bacteriorhodopsin are discussed. PMID- 14532281 TI - Proteomics analysis of rat brain postsynaptic density. Implications of the diverse protein functional groups for the integration of synaptic physiology. AB - The postsynaptic density contains multiple protein complexes that together relay the presynaptic neurotransmitter input to the activation of the postsynaptic neuron. In the present study we took two independent proteome approaches for the characterization of the protein complement of the postsynaptic density, namely 1) two-dimensional gel electrophoresis separation of proteins in conjunction with mass spectrometry to identify the tryptic peptides of the protein spots and 2) isolation of the trypsin-digested sample that was labeled with isotope-coded affinity tag, followed by liquid chromatography-tandem mass spectrometry for the partial separation and identification of the peptides, respectively. Functional grouping of the identified proteins indicates that the postsynaptic density is a structurally and functionally complex organelle that may be involved in a broad range of synaptic activities. These proteins include the receptors and ion channels for glutamate neurotransmission, proteins for maintenance and modulation of synaptic architecture, sorting and trafficking of membrane proteins, generation of anaerobic energy, scaffolding and signaling, local protein synthesis, and correct protein folding and breakdown of synaptic proteins. Together, these results imply that the postsynaptic density may have the ability to function (semi-) autonomously and may direct various cellular functions in order to integrate synaptic physiology. PMID- 14532282 TI - HCF-1 functions as a coactivator for the zinc finger protein Krox20. AB - HCF-1 is a transcriptional cofactor required for activation of herpes simplex virus immediate-early genes by VP16 as well as less clearly defined roles in cell proliferation, cytokinesis, and spliceosome formation. It is expressed as a large precursor that undergoes proteolysis to yield two subunits that remain stably associated. VP16 uses a degenerate 4-amino acid sequence, known as the HCF binding motif, to bind to a six-bladed beta-propeller domain at the N terminus of HCF-1. Functional HCF-binding motifs are also found in LZIP and Zhangfei, two cellular bZIP transcription factors of unknown function. Here we show that the HCF-binding motif occurs in a wide spectrum of DNA-binding proteins and transcriptional cofactors. Three well characterized examples were further analyzed for their ability to use HCF-1 as a coactivator. Krox20, a zinc finger transcription factor required for Schwann cell differentiation, and E2F4, a cell cycle regulator, showed a strong requirement for functional HCF-1 to activate transcription. In contrast, activation by estrogen receptor-alpha did not display HCF dependence. In Krox20, the HCF-binding motif lies within the N-terminal activation domain and mutation of this sequence diminishes both transactivation and association with the HCF-1 beta-propeller. The activation domain in the C terminal subunit of HCF-1 contributes to activation by Krox20, possibly through recruitment of p300. These results suggest that HCF-1 is recruited by many different classes of cellular transcription factors and is therefore likely to be required for a variety of cellular processes including cell cycle progression and development. PMID- 14532283 TI - The CD81 tetraspanin facilitates instantaneous leukocyte VLA-4 adhesion strengthening to vascular cell adhesion molecule 1 (VCAM-1) under shear flow. AB - Leukocyte integrins must rapidly strengthen their binding to target endothelial sites to arrest rolling adhesions under physiological shear flow. We demonstrate that the integrin-associated tetraspanin, CD81, regulates VLA-4 and VLA-5 adhesion strengthening in monocytes and primary murine B cells. CD81 strengthens multivalent VLA-4 contacts within subsecond integrin occupancy without altering intrinsic adhesive properties to low density ligand. CD81 facilitates both VLA-4 mediated leukocyte rolling and arrest on VCAM-1 under shear flow as well as VLA-5 dependent adhesion to fibronectin during short stationary contacts. CD81 also augments VLA-4 avidity enhancement induced by either chemokine-stimulated Gi proteins or by protein kinase C activation, although it is not required for Gi protein or protein kinase C signaling activities. In contrast to other proadhesive integrin-associated proteins, CD81-promoted integrin adhesiveness does not require its own ligand occupancy or ligation. These results provide the first demonstration of an integrin-associated transmembranal protein that facilitates instantaneous multivalent integrin occupancy events that promote leukocyte adhesion to an endothelial ligand under shear flow. PMID- 14532284 TI - Latent membrane protein 1 of Epstein-Barr virus stimulates processing of NF-kappa B2 p100 to p52. AB - Recent studies have identified a limited number of cellular receptors that can stimulate an alternative NF-kappa B activation pathway that depends upon the inducible processing of NF-kappa B2 p100 to p52. Here it is shown that the latent membrane protein (LMP)-1 of Epstein-Barr virus can trigger this signaling pathway in both B cells and epithelial cells. LMP1-induced p100 processing, which is mediated by the proteasome and is dependent upon de novo protein synthesis, results in the nuclear translocation of p52.RelB dimers. Previous studies have established that LMP1 also stimulates the canonical NF-kappa B-signaling pathway that triggers phosphorylation and degradation of I kappa B alpha. Interestingly, LMP1 activation of these two NF-kappa B pathways is shown here to require distinct regions of the LMP1 C-terminal cytoplasmic tail. Thus, C-terminal activating region 1 is required for maximal triggering of p100 processing but is largely dispensable for stimulation of I kappa B alpha phosphorylation. In contrast, C-terminal-activating region 2 is critical for maximal LMP1 triggering of I kappa B alpha phosphorylation and up-regulation of p100 levels but does not contribute to activation of p100 processing. Because p100 deletion mutants that constitutively produce p52 oncogenically transform fibroblasts in vitro, it is likely that stimulation of p100 processing by LMP1 will play an important role in its transforming function. PMID- 14532285 TI - PKC epsilon is a unique regulator for hsp90 beta gene in heat shock response. AB - An early event in cellular heat shock response is the transmittance of stress signals from the cell surface into the nuclei, resulting in the induction of heat shock proteins (Hsps). Protein kinase C (PKC) has been implicated as a key player in transducing stress signals. However, mechanism(s) by which PKC regulates heat shock-induced events remains largely unknown. Here we present data that pan-PKC inhibitor GF109203X, but not classic PKC inhibitor Go6976, specifically repressed heat shock-induced accumulation of mRNA as well as promoter activity of hsp90 beta, but not hsp90 alpha, in Jurkat cells. Subcellular fractionation studies revealed that heat shock exclusively induced PKC-epsilon membrane translocation. Consistently, expression of a constitutively active PKC-epsilon(A159E) resulted in an enhanced promoter activity of hsp90 beta upon heat shock, whereas a dominant-negative PKC-epsilon(K437R) abolished this effect. In contrast, constitutively active-PKC-alpha or dominant-negative-PKC-alpha had no effects on heat shock induction of the gene. The effect of PKC-epsilon on hsp90 beta expression seems to be stimuli-specific, as phorbol myristate acetate-mediated hsp90 beta expression was PKC-epsilon-independent. We conclude that PKC-epsilon is specifically required in the signaling pathway leading to the induction of hsp90 beta gene in response to heat shock. PMID- 14532286 TI - A role for tumor necrosis factor receptor-2 and receptor-interacting protein in programmed necrosis and antiviral responses. AB - Members of the tumor necrosis factor (TNF) receptor (TNFR) superfamily are potent regulators of apoptosis, a process that is important for the maintenance of immune homeostasis. Recent evidence suggests that TNFR-1 and Fas and TRAIL receptors can also trigger an alternative form of cell death that is morphologically distinct from apoptosis. Because distinct molecular components including the serine/threonine protein kinase receptor-interacting protein (RIP) are required, we have referred to this alternative form of cell death as "programmed necrosis." We show that TNFR-2 signaling can potentiate programmed necrosis via TNFR-1. When cells were pre-stimulated through TNFR-2 prior to subsequent activation of TNFR-1, enhanced cell death and recruitment of RIP to the TNFR-1 complex were observed. However, TNF-induced programmed necrosis was normally inhibited by caspase-8 cleavage of RIP. To ascertain the physiological significance of RIP and programmed necrosis, we infected Jurkat cells with vaccinia virus (VV) and found that VV-infected cells underwent programmed necrosis in response to TNF, but deficiency of RIP rescued the infected cells from TNF-induced cytotoxicity. Moreover, TNFR-2-/- mice exhibited reduced inflammation in the liver and defective viral clearance during VV infection. Interestingly, death effector domain-containing proteins such as MC159, E8, K13, and cellular FLIP, but not the apoptosis inhibitors Bcl-xL, p35, and XIAP, potently suppressed programmed necrosis. Thus, TNF-induced programmed necrosis is facilitated by TNFR-2 signaling and caspase inhibition and may play a role in controlling viral infection. PMID- 14532287 TI - Electrophysiological analysis of the mutated Na,K-ATPase cation binding pocket. AB - Na,K-ATPase mediates net electrogenic transport by extruding three Na+ ions and importing two K+ ions across the plasma membrane during each reaction cycle. We mutated putative cation coordinating amino acids in transmembrane hairpin M5-M6 of rat Na,K-ATPase: Asp776 (Gln, Asp, Ala), Glu779 (Asp, Gln, Ala), Asp804 (Glu, Asn, Ala), and Asp808 (Glu, Asn, Ala). Electrogenic cation transport properties of these 12 mutants were analyzed in two-electrode voltage-clamp experiments on Xenopus laevis oocytes by measuring the voltage dependence of K+-stimulated stationary currents and pre-steady-state currents under electrogenic Na+/Na+ exchange conditions. Whereas mutants D804N, D804A, and D808A hardly showed any Na+/K+ pump currents, the other constructs could be classified according to the [K+] and voltage dependence of their stationary currents; mutants N776A and E779Q behaved similarly to the wild-type enzyme. Mutants E779D, E779A, D808E, and D808N had in common a decreased apparent affinity for extracellular K+. Mutants N776Q, N776D, and D804E showed large deviations from the wild-type behavior; the currents generated by mutant N776D showed weaker voltage dependence, and the current-voltage curves of mutants N776Q and D804E exhibited a negative slope. The apparent rate constants determined from transient Na+/Na+ exchange currents are rather voltage-independent and at potentials above -60 mV faster than the wild type. Thus, the characteristic voltage-dependent increase of the rate constants at hyperpolarizing potentials is almost absent in these mutants. Accordingly, dislocating the carboxamide or carboxyl group of Asn776 and Asp804, respectively, decreases the extracellular Na+ affinity. PMID- 14532288 TI - Role of ADMIDAS cation-binding site in ligand recognition by integrin alpha 5 beta 1. AB - Integrin-ligand interactions are regulated in a complex manner by divalent cations, and multiple cation-binding sites are found in both alpha and beta integrin subunits. A key cation-binding site that lies in the beta subunit A domain is known as the metal-ion dependent adhesion site (MIDAS). Recent x-ray crystal structures of integrin alpha V beta 3 have identified a novel cation binding site in this domain, known as the ADMIDAS (adjacent to MIDAS). The role of this novel site in ligand recognition has yet to be elucidated. Using the interaction between alpha 5 beta 1 and fibronectin as a model system, we show that mutation of residues that form the ADMIDAS site inhibits ligand binding but this effect can be partially rescued by the use of activating monoclonal antibodies. The ADMIDAS mutants had decreased expression of activation epitopes recognized by 12G10, 15/7, and HUTS-4, suggesting that the ADMIDAS is important for stabilizing the active conformation of the integrin. Consistent with this suggestion, the ADMIDAS mutations markedly increased the dissociation rate of the integrin-fibronectin complex. Mutation of the ADMIDAS residues also reduced the allosteric inhibition of Mn2+-supported ligand binding by Ca2+, suggesting that the ADMIDAS is a Ca2+-binding site involved in the inhibition of Mn2+-supported ligand binding. Mutations of the ADMIDAS site also perturbed transduction of a conformational change from the MIDAS through the C-terminal helix region of the beta A domain to the underlying hybrid domain, implying an important role for this site in receptor signaling. PMID- 14532289 TI - Heterodimerization of substance P and mu-opioid receptors regulates receptor trafficking and resensitization. AB - The micro-opioid receptor (MOR1) and the substance P receptor (NK1) coexist and functionally interact in nociceptive brain regions; however, a molecular basis for this interaction has not been established. Using coimmunoprecipitation and bioluminescence resonance energy transfer (BRET), we show that MOR1 and NK1 can form heterodimers in HEK 293 cells coexpressing the two receptors. Although NK1 MOR1 heterodimerization did not substantially change the ligand binding and signaling properties of these receptors, it dramatically altered their internalization and resensitization profile. Exposure of the NK1-MOR1 heterodimer to the MOR1-selective ligand [D-Ala2,Me-Phe4,Gly5-ol]enkephalin (DAMGO) promoted cross-phosphorylation and cointernalization of the NK1 receptor. Conversely, exposure of the NK1-MOR1 heterodimer to the NK1-selective ligand substance P (SP) promoted cross-phosphorylation and cointernalization of the MOR1 receptor. In cells expressing MOR1 alone, beta-arrestin directs the receptors to clathrin coated pits, but does not internalize with the receptor. In cells expressing NK1 alone, beta-arrestin internalizes with the receptor into endosomes. Interestingly, in cells coexpressing MOR1 and NK1 both DAMGO and SP induced the recruitment of beta-arrestin to the plasma membrane and cointernalization of NK1 MOR1 heterodimers with beta-arrestin into the same endosomal compartment. Consequently, resensitization of MOR1-dependent receptor functions was severely delayed in coexpressing cells as compared with cells expressing MOR1 alone. Together, our findings indicate that MOR1 by virtue of its physical interaction with NK1 is sequestered via an endocytotic pathway with delayed recycling and resensitization kinetics. PMID- 14532290 TI - Down-regulation of Sp1 activity through modulation of O-glycosylation by treatment with a low glucose mimetic, 2-deoxyglucose. AB - 2-Deoxyglucose (2-DG), a nonmetabolizable glucose analogue, blocks glycolysis at the phosphohexose isomerase step and has been frequently used as a glucose starvation mimetic in studies of a wide variety of physiological dysfuctions. However, the effect of 2-DG on protein glycosylation and related signal pathways has not been investigated in depth. In HeLa, an HPV18-positive cervical carcinoma line, 2-DG treatment down-regulates human papillomavirus early gene transcription. This down-regulation was also achieved by low glucose supply or hypoxia, suggesting that this is a response commonly modulated by cellular glucose or energy level. We investigated how 2-DG and low glucose affect transcriptional activity. Human papillomavirus gene transcription was only marginally affected by the inhibition of ATP synthesis or the supplementation of pyruvate to 2-DG-treated cells, suggesting that poor ATP generation is involved only to a limited extent. 2-DG treatment also inhibited activation of p21 WAF1 promoter, which is controlled by p53 and/or Sp1. In a reporter assay using p21 WAF1 promoter constructs, 2-DG exerted a strong inhibitory effect on Sp1 activity. DNA binding activity of Sp1 in 2-DG-treated HeLa cells was intact, whereas it was severely impaired in cells incubated in a low glucose medium or in hypoxic condition. Unexpectedly, Sp1 was heavily modified with GlcNAc in 2-DG treated cells, which is at least partially attributed to the inhibitory effect of 2-DG on N-acetyl-beta-D-glucosaminidase activity. Our results suggest that 2-DG, like low glucose or hypoxic condition, down-regulates Sp1 activity, but through hyper-GlcNAcylation instead of hypo-GlcNAcylation. PMID- 14532291 TI - Role of the conserved SRLFDQFFG region of alpha-crystallin, a small heat shock protein. Effect on oligomeric size, subunit exchange, and chaperone-like activity. AB - Small heat shock proteins (sHsps) are necessary for several cellular functions and in stress tolerance. Most sHsps are oligomers; intersubunit interactions leading to changes in oligomeric structure and exposure of specific regions may modulate their functioning. Many sHsps, including alpha A- and alpha B crystallin, contain a well conserved SRLFDQFFG sequence motif in the N-terminal region. Sequence-based prediction shows that it exhibits helical propensity with amphipathic character, suggesting that it plays a critical role in the structure and function of alpha-crystallins. In order to investigate the role of this motif in the structure and function of sHsps, we have made constructs deleting this sequence from alpha A- and alpha B-crystallin, overexpressed, purified, and studied these engineered proteins. Circular dichroism spectroscopic studies show changes in tertiary and secondary structure on deletion of the sequence. Glycerol density gradient centrifugation and dynamic light scattering studies show that the multimeric size of the mutant proteins is significantly reduced, indicating a role for this motif in higher order organization of the subunits. Both deletion mutants exhibit similar oligomeric size and increased chaperone-like activity. Urea-induced denaturation study shows that the SRLFDQFFG sequence contributes significantly to the structural stability. Fluorescence resonance energy transfer studies show that the rate of exchange of the subunits in the alpha Adel crystallin oligomer is higher compared with that in the alpha A-crystallin oligomer, suggesting that this region contributes to the oligomer dynamics in addition to the higher order assembly and structural stability. Thus, our study shows that the SRLFDQFFG sequence is one of the critical motifs in structure function regulation of alpha A- and alpha B-crystallin. PMID- 14532292 TI - Mullerian inhibiting substance promotes interferon gamma-induced gene expression and apoptosis in breast cancer cells. AB - This report demonstrates that in addition to interferons and cytokines, members of the TGF beta superfamily such as Mullerian inhibiting substance (MIS) and activin A also regulate IRF-1 expression. MIS induced IRF-1 expression in the mammary glands of mice in vivo and in breast cancer cells in vitro and stimulation of IRF-1 by MIS was dependent on activation of the NF kappa B pathway. In the rat mammary gland, IRF-1 expression gradually decreased during pregnancy and lactation but increased at involution. In breast cancer, the IRF-1 protein was absent in 13% of tumors tested compared with matched normal glands. Consistent with its growth suppressive activity, expression of IRF-1 in breast cancer cells induced apoptosis. Treatment of breast cancer cells with MIS and interferon gamma (IFN-gamma) co-stimulated IRF-1 and CEACAM1 expression and synergistic induction of CEACAM1 by a combination of MIS and IFN-gamma was impaired by antisense IRF-1 expression. Furthermore, a combination of IFN-gamma and MIS inhibited the growth of breast cancer cells to a greater extent than either one alone. Both reagents alone significantly decreased the fraction of cells in the S-phase of the cell cycle, an effect not enhanced when they were used in combination. However, MIS promoted IFN-gamma-induced apoptosis demonstrating a functional interaction between these two classes of signaling molecules in regulation of breast cancer cell growth. PMID- 14532293 TI - 5'-AMP-activated protein kinase controls insulin-containing secretory vesicle dynamics. AB - Changes in 5'-AMP-activated protein kinase (AMPK) activity have recently been implicated in the control of insulin secretion by glucose (da Silva Xavier, G., Leclerc, I., Varadi, A., Tsuboi, T., Moule, S. K., and Rutter, G. A. (2003) Biochem. J. 371, 761-774). Here, we examine the possibility that activation of AMPK may regulate distal steps in insulin secretion, including vesicle movement and fusion with the plasma membrane. Vesicle dynamics were imaged in single pancreatic MIN6 beta-cells expressing lumen-targeted pH-insensitive yellow fluorescent protein, neuropeptide Y.Venus, or monomeric red fluorescent protein by total internal reflection fluorescence and Nipkow disc confocal microscopy. Overexpression of a truncated, constitutively active form of AMPK (AMPKalpha1, 1 312, T172D; AMPK CA), inhibited glucose-stimulated (30 versus 3.0 mM) vesicle movements, and decreased the number of vesicles docked or fusing at the plasma membrane, while having no effect on the kinetics of individual secretory events. Expression of the activated form of AMPK also prevented dispersal of the cortical actin network at high glucose concentrations. Monitored in permeabilized cells, where the effects of AMPK CA on glucose metabolism and ATP synthesis were bypassed, AMPK CA inhibited Ca2+ and ATP-induced insulin secretion, and decreased ATP-dependent vesicle movements. These findings suggest that components of the vesicle transport network, including vesicle-associated motor proteins, may be targets of AMPK in beta-cells, dephosphorylation of which is required for vesicle mobilization at elevated glucose concentrations. PMID- 14532294 TI - Cis-acting elements stimulating kinetoplastid guide RNA-directed editing. AB - The coding sequence of several mitochondrial mRNAs of the kinetoplastid protozoa is created through the insertion and deletion of specific uridylates. The editing reactions are required to be highly specific in order to ensure that functional open reading frames are created in edited mRNAs and that potentially deleterious modification of normally nonedited sequence does not occur. Selection amplification and mutagenesis were previously used to identify the optimal sequence requirements for in vitro editing. There is, however, a minority of natural editing sites with suboptimal sequence. Several cis-acting elements, obtained from an in vitro selection, are described here that are able to compensate for a suboptimal editing site. An A + U sequence element within the 5' untranslated region of cytochrome b mRNA from Leishmania tarentolae is also demonstrated to function as a cis-acting guide RNA and is postulated to compensate for a suboptimal editing site in vivo. Two proteins within an enriched editing extract are UV-cross-linked to two different in vitro selected editing substrates more efficiently than poorly edited RNAs. The results suggest that these proteins contribute to the specificity of the editing reaction. PMID- 14532295 TI - RasGRP1 represents a novel non-protein kinase C phorbol ester signaling pathway in mouse epidermal keratinocytes. AB - The mouse skin model of carcinogenesis has been instrumental in our appreciation of the multistage nature of carcinogenesis. In this system, tumor promotion is a critical step in the generation of tumors and is usually achieved by treatment with the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA). Although it is generally assumed that protein kinase C (PKC) is the sole receptor for TPA in this system, we sought to evaluate whether non-PKC pathways could also contribute to the effects of phorbol esters in skin. We documented expression of the high affinity non-PKC phorbol ester receptor and Ras activator RasGRP1 in mouse primary keratinocytes. Overexpression of RasGRP1 in keratinocytes increased the level of active GTP-loaded Ras. TPA treatment further elevated this Ras activation in a PKC-independent manner and induced the translocation and down regulation of RasGRP1. Overexpression of RasGRP1 in keratinocytes also caused apoptosis. Finally, induction of keratinocyte differentiation by elevation of extracellular calcium suppressed expression of endogenous RasGRP1, whereas overexpression of RasGRP1 inhibited expression of the differentiation markers keratins 1 and 10 induced by high calcium in the medium. Taken together, our results demonstrate that RasGRP1 is an additional diacylglycerol/phorbol ester receptor in epidermal keratinocytes and suggest that activation of this novel receptor may contribute to some of the phorbol ester- and Ras-mediated effects in mouse epidermis. PMID- 14532296 TI - Fibrillogenic amylin evokes islet beta-cell apoptosis through linked activation of a caspase cascade and JNK1. AB - Fibrillogenic human amylin elicits pancreatic beta-cell apoptosis that may contribute to development of type-2 diabetes. Here, we demonstrated that activation of a caspase cascade is necessary for induction of apoptosis by fibrillogenic amylin variants in two pancreatic beta-cell lines. Human amylin, as well as truncated 8-37human amylin, evoked sequential activation of caspases-8 and -3, and apoptosis, whereas non-beta-sheet forming and non-fibrillogenic homologs, such as [25,28,29triprolyl]human amylin, did not, implying that the beta-sheet conformer is required for human amylin-induced caspase activation. Significant inhibition of apoptosis was evoked by a selective caspase-1 inhibitor, indicating that caspase-1 is also essential for activation of the caspase cascade. Furthermore, we showed that specific jnk1 antisense oligonucleotides, which suppress phospho-JNK1 expression, effectively decreased human amylin-induced activation of c-Jun. Studies of the interplay between the caspase cascade and the JNK pathway showed that both apoptosis and caspase-3 activation were suppressed by treatment with a JNK inhibitor and by transfection of antisense jnk1 oligonucleotides or antisense-c-jun, whereas a selective inhibitor of caspases-1 and -3 prevented apoptosis but not c-Jun activation. Thus, the JNK1 activation preceded activation of caspases-1 and -3. However, selective JNK inhibition had no effect on caspase-8 activation, and selective caspase-8 inhibition only partially suppressed apoptosis and c-Jun activation, indicating that caspase-8 may partially act upstream of the JNK pathway. Our studies demonstrate a functional interaction of a caspase cascade and JNK1. Fibrillogenic amylin can evoke a JNK1-mediated apoptotic pathway, which is partially dependent and partially independent of caspase-8, and in which caspase 3 acts as a common downstream effector. PMID- 14532297 TI - A novel human cytochrome P450, CYP26C1, involved in metabolism of 9-cis and all trans isomers of retinoic acid. AB - Retinoids are potent regulators of cell proliferation, cell differentiation, and morphogenesis and are important therapeutic agents in oncology and dermatology. The gene regulatory activity of endogenous retinoids is effected primarily by retinoic acid isomers (all-trans and 9-cis) that are synthesized from retinaldehyde precursors in a broad range of tissues and act as ligands for nuclear retinoic acid receptors. The catabolism of all-trans-retinoic acid (atRA) is an important mechanism of controlling RA levels in cell and tissues. We have previously identified two cytochrome P450s, P450RAI-1 and P450RAI-2 (herein named CYP26A1 and CYP26B1), which were shown to be responsible for catabolism of atRA both in the embryo and the adult. In this report, we describe the identification, molecular cloning, and substrate characterization of a third member of the CYP26 family, named CYP26C1. Transiently transfected cells expressing CYP26C1 convert atRA to polar water-soluble metabolites similar to those generated by CYP26A1 and -B1. Competition studies with all-trans, 13-cis, and 9-cis isomers of retinoic acid demonstrated that atRA was the preferred substrate for CYP26C1. Although CYP26C1 shares extensive sequence similarity with CYP26A1 and CYP26B1, its catalytic activity appears distinct from those of other CYP26 family members. Specifically, CYP26C1 can also recognize and metabolize 9-cis-RA and is much less sensitive than the other CYP26 family members to the inhibitory effects of ketoconazole. CYP26C1 is not widely expressed in the adult but is inducible by RA in HPK1a, transformed human keratinocyte cell lines. This third CYP26 member may play a specific role in catabolizing both all-trans and 9-cis isomers of RA. PMID- 14532298 TI - Researchers probe anemia-heart failure link. PMID- 14532299 TI - Breast cancer racial gap examined: no easy answers to explain disparities in survival. PMID- 14532300 TI - Telemedicine eyed for mental health services: approach could widen access for older patients. PMID- 14532304 TI - Patients with viral infections who demand antibiotics. PMID- 14532305 TI - The value of diagnostic tests for low back pain. PMID- 14532306 TI - Patients with viral infections who demand antibiotics. PMID- 14532307 TI - The value of diagnostic tests for low back pain. PMID- 14532308 TI - The value of diagnostic tests for low back pain. PMID- 14532309 TI - Computer use and risk of carpal tunnel syndrome. PMID- 14532310 TI - Computer use and risk of carpal tunnel syndrome. PMID- 14532311 TI - Causes of renal failure in patients with type 2 diabetes mellitus. PMID- 14532312 TI - Causes of renal failure in patients with type 2 diabetes mellitus. PMID- 14532313 TI - Methyl isocyanate exposure and growth patterns of adolescents in Bhopal. PMID- 14532314 TI - Association of low-level ozone and fine particles with respiratory symptoms in children with asthma. AB - CONTEXT: Exposure to ozone and particulate matter of 2.5 microm or less (PM2.5) in air at levels above current US Environmental Protection Agency (EPA) standards is a risk factor for respiratory symptoms in children with asthma. OBJECTIVE: To examine simultaneous effects of ozone and PM2.5 at levels below EPA standards on daily respiratory symptoms and rescue medication use among children with asthma. DESIGN, SETTING, AND PARTICIPANTS: Daily respiratory symptoms and medication use were examined prospectively for 271 children younger than 12 years with physician diagnosed, active asthma residing in southern New England. Exposure to ambient concentrations of ozone and PM2.5 from April 1 through September 30, 2001, was assessed using ozone (peak 1-hour and 8-hour) and 24-hour PM2.5. Logistic regression analyses using generalized estimating equations were performed separately for maintenance medication users (n = 130) and nonusers (n = 141). Associations between pollutants (adjusted for temperature, controlling for same- and previous-day levels) and respiratory symptoms and use of rescue medication were evaluated. MAIN OUTCOME MEASURES: Respiratory symptoms and rescue medication use recorded on calendars by subjects' mothers. RESULTS: Mean (SD) levels were 59 (19) ppb (1-hour average) and 51 (16) ppb (8-hour average) for ozone and 13 (8) microg/m3 for PM2.5. In copollutant models, ozone level but not PM2.5 was significantly associated with respiratory symptoms and rescue medication use among children using maintenance medication; a 50-ppb increase in 1-hour ozone was associated with increased likelihood of wheeze (by 35%) and chest tightness (by 47%). The highest levels of ozone (1-hour or 8-hour averages) were associated with increased shortness of breath and rescue medication use. No significant, exposure-dependent associations were observed for any outcome by any pollutant among children who did not use maintenance medication. CONCLUSION: Asthmatic children using maintenance medication are particularly vulnerable to ozone, controlling for exposure to fine particles, at levels below EPA standards. PMID- 14532315 TI - Excess length of stay, charges, and mortality attributable to medical injuries during hospitalization. AB - CONTEXT: Although medical injuries are recognized as a major hazard in the health care system, little is known about their impact. OBJECTIVE: To assess excess length of stay, charges, and deaths attributable to medical injuries during hospitalization. DESIGN, SETTING, AND PATIENTS: The Agency for Healthcare Research and Quality (AHRQ) Patient Safety Indicators (PSIs) were used to identify medical injuries in 7.45 million hospital discharge abstracts from 994 acute-care hospitals across 28 states in 2000 in the AHRQ Healthcare Cost and Utilization Project Nationwide Inpatient Sample database. MAIN OUTCOME MEASURES: Length of stay, charges, and mortality that were recorded in hospital discharge abstracts and were attributable to medical injuries according to 18 PSIs. RESULTS: Excess length of stay attributable to medical injuries ranged from 0 days for injury to a neonate to 10.89 days for postoperative sepsis, excess charges ranged from 0 dollar for obstetric trauma (without vaginal instrumentation) to 57 727 dollars for postoperative sepsis, and excess mortality ranged from 0% for obstetric trauma to 21.96% for postoperative sepsis (P<.001). Following postoperative sepsis, the second most serious event was postoperative wound dehiscence, with 9.42 extra days in the hospital, 40 323 dollars in excess charges, and 9.63% attributable mortality. Infection due to medical care was associated with 9.58 extra days, 38 656 dollars in excess charges, and 4.31% attributable mortality. CONCLUSION: Some injuries incurred during hospitalization pose a significant threat to patients and costs to society, but the impact of such injury is highly variable. PMID- 14532317 TI - Lifetime risk for diabetes mellitus in the United States. AB - CONTEXT: Although diabetes mellitus is one of the most prevalent and costly chronic diseases in the United States, no estimates have been published of individuals' average lifetime risk of developing diabetes. OBJECTIVE: To estimate age-, sex-, and race/ethnicity-specific lifetime risk of diabetes in the cohort born in 2000 in the United States. DESIGN, SETTING, AND PARTICIPANTS: Data from the National Health Interview Survey (1984-2000) were used to estimate age-, sex , and race/ethnicity-specific prevalence and incidence in 2000. US Census Bureau data and data from a previous study of diabetes as a cause of death were used to estimate age-, sex-, and race/ethnicity-specific mortality rates for diabetic and nondiabetic populations. MAIN OUTCOME MEASURES: Residual (remaining) lifetime risk of diabetes (from birth to 80 years in 1-year intervals), duration with diabetes, and life-years and quality-adjusted life-years lost from diabetes. RESULTS: The estimated lifetime risk of developing diabetes for individuals born in 2000 is 32.8% for males and 38.5% for females. Females have higher residual lifetime risks at all ages. The highest estimated lifetime risk for diabetes is among Hispanics (males, 45.4% and females, 52.5%). Individuals diagnosed as having diabetes have large reductions in life expectancy. For example, we estimate that if an individual is diagnosed at age 40 years, men will lose 11.6 life-years and 18.6 quality-adjusted life-years and women will lose 14.3 life years and 22.0 quality-adjusted life-years. CONCLUSIONS: For individuals born in the United States in 2000, the lifetime probability of being diagnosed with diabetes mellitus is substantial. Primary prevention of diabetes and its complications are important public health priorities. PMID- 14532316 TI - Treatment of corticosteroid-responsive autoimmune inner ear disease with methotrexate: a randomized controlled trial. AB - CONTEXT: A number of therapies have been proposed for the long-term management of corticosteroid-responsive, rapidly progressive, bilateral sensorineural hearing loss (autoimmune inner ear disease [AIED]). Methotrexate has emerged as the benchmark agent but has not been rigorously evaluated for hearing improvement in patients with AIED. OBJECTIVE: To assess the efficacy of long-term methotrexate in maintaining hearing improvements achieved with glucocorticoid (prednisone) therapy in patients with AIED. DESIGN, SETTING, AND PARTICIPANTS: A randomized, double-blind, placebo-controlled trial conducted from February 3, 1998, to November 5, 2001, of 67 patients with rapidly progressive, bilateral sensorineural hearing loss at 10 tertiary care centers in the United States. INTERVENTION: Randomization to either oral methotrexate (15 to 20 mg/wk; n = 33) or placebo (n = 34), in combination with an 18-week prednisone taper. Follow-up examinations, including audiometric evaluation, were performed at 4, 8, 12, 24, 36, 48, and 52 weeks, or until hearing loss was documented. MAIN OUTCOME MEASURE: Maintenance of hearing improvement achieved from prednisone treatment. RESULTS: Sixty-seven patients (57.8%) enrolled in the prednisone challenge experienced hearing improvement. Twenty-five patients (37%) experienced hearing improvements in both ears. Of the individuals who reached study end points, 24 (80%) of 30 end points were because of measured hearing loss in the methotrexate group and 29 (93.5%) of 31 end points were because of measured hearing loss in the placebo group (P =.15). Methotrexate was no more effective than placebo in maintaining the hearing improvement achieved with prednisone treatment (hazard ratio, 1.31; 95% confidence interval, 0.79-2.17; P =.30). CONCLUSION: Methotrexate does not appear to be effective in maintaining the hearing improvement achieved with prednisone therapy in patients with AIED. PMID- 14532318 TI - Mortality benefit of immediate revascularization of acute ST-segment elevation myocardial infarction in patients with contraindications to thrombolytic therapy: a propensity analysis. AB - CONTEXT: There are no definitive recommendations for the management of acute myocardial infarction (AMI) in patients with ST-segment elevation who have contraindications to thrombolytic therapy. It is not clear whether, and the extent to which, immediate mechanical reperfusion (IMR) reduces in-hospital mortality in this population. OBJECTIVE: To determine whether IMR (defined as percutaneous coronary intervention or coronary artery bypass graft surgery) is associated with a mortality benefit in patients with acute ST-segment elevation AMI who are eligible for IMR but have contraindications to thrombolytic therapy. DESIGN, SETTING, AND PATIENTS: From June 1994 to January 2003, the National Registry of Myocardial Infarction 2, 3, and 4 enrolled 1 799 704 patients with AMI. A total of 19 917 patients with acute ST-segment elevation were eligible for IMR but had thrombolytic contraindications after excluding patients who were transferred in from or out to other facilities, patients who received intracoronary thrombolytics, and those who received no medications within 24 hours of arrival. MAIN OUTCOME MEASURE: In-hospital mortality. RESULTS: Of the 19 917 patients, 4705 patients (23.6%) received IMR and 5173 patients (25.9%) died. In-hospital mortality rates in the IMR and non-IMR treated groups in the unadjusted analysis were 11.1%, representing 521 of 4705 patients, and 30.6%, representing 4652 of 15 212 patients, respectively, for a risk reduction of 63.7% (odds ratio [OR], 0.28; 95% confidence interval [CI], 0.26-0.31). In a further analysis using a propensity matching score to reduce the effects of bias, 3905 patients who received IMR remained at lower risk for in-hospital mortality than 3905 matched patients (10.9% vs 20.1%, respectively, for a risk reduction of 45.8%; OR, 0.48; 95% CI, 0.43-0.55). Following a second logistic model applied to the matched groups to adjust for residual differences, a significant treatment effect persisted (OR, 0.64; 95% CI, 0.56-0.75). CONCLUSIONS: In this population, IMR was associated with a reduced risk of in-hospital mortality after appropriate adjustments. Of those we studied who were eligible for IMR, 15 212 patients (76.4%) did not receive it. These results suggest that using IMR in patients with acute ST-segment elevation AMI and contraindications to thrombolytics should be strongly considered. PMID- 14532319 TI - Safety of patients isolated for infection control. AB - CONTEXT: Hospital infection control policies that use patient isolation prevent nosocomial transmission of infectious diseases, but may inadvertently lead to patient neglect and errors. OBJECTIVE: To examine the quality of medical care received by patients isolated for infection control. DESIGN, SETTING, AND PATIENTS: We identified consecutive adults who were isolated for methicillin resistant Staphylococcus aureus colonization or infection at 2 large North American teaching hospitals: a general cohort (patients admitted with all diagnoses between January 1, 1999, and January 1, 2000; n = 78); and a disease specific cohort (patients admitted with a diagnosis of congestive heart failure between January 1, 1999, and July 1, 2002; n = 72). Two matched controls were selected for each isolated patient (n = 156 general cohort controls and n = 144 disease-specific cohort controls). MAIN OUTCOME MEASURES: Quality-of-care measures encompassing processes, outcomes, and satisfaction. Adjustments for study cohort and patient demographic, hospital, and clinical characteristics were conducted using multivariable regression. RESULTS: Isolated and control patients generally had similar baseline characteristics; however, isolated patients were twice as likely as control patients to experience adverse events during their hospitalization (31 vs 15 adverse events per 1000 days; P<.001). This difference in adverse events reflected preventable events (20 vs 3 adverse events per 1000 days; P<.001) as opposed to nonpreventable events (11 vs 12 adverse events per 1000 days; P =.98). Isolated patients were also more likely to formally complain to the hospital about their care than control patients (8% vs 1%; P<.001), to have their vital signs not recorded as ordered (51% vs 31%; P<.001), and more likely to have days with no physician progress note (26% vs 13%; P<.001). No differences in hospital mortality were observed for the 2 groups (17% vs 10%; P =.16). CONCLUSION: Compared with controls, patients isolated for infection control precautions experience more preventable adverse events, express greater dissatisfaction with their treatment, and have less documented care. PMID- 14532320 TI - Obstructive sleep apnea: implications for cardiac and vascular disease. AB - CONTEXT: Obstructive sleep apnea (OSA) has been increasingly implicated in the initiation and progression of cardiovascular diseases. OBJECTIVE: To systematically review the interactions of OSA with cardiovascular pathophysiology and diseases. DATA SOURCES AND STUDY SELECTION: The MEDLINE database from January 1966 to March 2003 was searched using the Medical Subject Headings sleep, sleep apnea, obesity, hypertension, heart failure, cardiac arrhythmia, coronary artery disease, stroke, sympathetic activity, endothelium, inflammation, and continuous positive airway pressure (CPAP) to identify peer-reviewed studies of OSA. Priority was given to large prospective cohort studies and to randomized controlled trials. DATA EXTRACTION: We identified 154 original investigations and reviews of sleep-related breathing disorders. Data from these studies were examined for relevance and extracted by one of the authors. DATA SYNTHESIS: Approximately 1 in 5 adults has at least mild OSA (apnea-hypopnea index [ie, the number of apneic and hypopneic events per hour], 5-15), and 1 in 15 adults has at least moderate OSA (apnea-hypopnea index, 15-30). Repetitive apneic events disrupt the normal physiologic interactions between sleep and the cardiovascular system. Such sleep fragmentation, as well as abnormalities evident in patients with OSA (eg, increased sympathetic activation, vascular endothelial dysfunction, increased oxidative stress, inflammation, increased platelet aggregability, metabolic dysregulation), may be implicated in the initiation and progression of cardiac and vascular disease. Persuasive data implicate OSA in the development of hypertension, and OSA also may contribute to cardiac ischemia, congestive heart failure, cardiac arrhythmias, and perhaps also to cerebrovascular disease and stroke. CONCLUSIONS: Obstructive sleep apnea is common, readily diagnosed, and usually treatable. It frequently coexists undiagnosed in patients with cardiovascular disease, activates disease mechanisms known to elicit cardiac and vascular damage, and may be implicated in progression of cardiovascular disease and resistance to conventional therapeutic strategies. In the absence of definitive evidence from large-scale trials and a better understanding of potential cost-effectiveness, the likely benefits of diagnosis and treatment of OSA are presently best appraised on an individualized patient basis. PMID- 14532321 TI - Air pollution as an underappreciated cause of asthma symptoms. PMID- 14532322 TI - Looking for medical injuries where the light is bright. PMID- 14532323 TI - JAMA patient page. Health effects of ozone. PMID- 14532324 TI - A frameshifting mutation in CHRNE unmasks skipping of the preceding exon. AB - A frameshifting 7 bp deletion (epsilon553del7) in exon 7 of CHRNE encoding the acetylcholine receptor epsilon subunit, observed in seven congenital myasthenic syndrome patients, enhances expression of an aberrantly spliced transcript that skips the preceding 101 bp exon 6. To recapitulate the aberrant splicing, we cloned the entire CHRNE spanning 12 exons and 11 introns and expressed it in COS cells. Scanning mutagenesis revealed that epsilon553del7 does not disrupt an exonic splicing enhancer. Inhibition of protein synthesis and of nonsense mediated mRNA decay (NMD) by anisomycin shows that even wild-type CHRNE produces an exon 6-skipped transcript, and that even epsilon553del7-CHRNE yields a normally spliced transcript. Both transcripts, however, are degraded by NMD due to a premature stop codon. In contrast, the normally spliced transcript from wild type CHRNE and the exon 6-skipped transcript from epsilon553del7-CHRNE carry no premature stop codon and hence are immune to NMD. Optimization of splicing signals for exon 6 prevents it being skipped even in the presence of anisomycin and/or epsilon553del7, indicating that inherently weak splicing signals for exon 6 account for its skipping. We suggest that a similar mechanism probably operates in other genes in skipping of remote exons. The presence of weak splicing signals for exon 6 also prompted us to search for mutations in exon 6 that disrupt an exonic splicing enhancer. Indeed, we found that epsilonEF157V and epsilonE154X in exon 6, observed in two other patients, caused aberrant splicing of exon 6. PMID- 14532325 TI - Fragile X Mental Retardation protein determinants required for its association with polyribosomal mRNPs. AB - Fragile X Mental Retardation protein (FMRP) is an RNA-binding protein that contains multiple domains with apparently differential affinity to mRNA and to the ribonucleotide homopolymer poly(G). Attempts have been made to map the RNA binding sites along the protein sequence with a view to determining which of the KH1, KH2 and RGG domains are required to recognize and bind to RNA. While these studies have greatly contributed to the delineation of domains that bind homopolymers or mRNA in vitro, little is known concerning their implications in FMRP function(s) in vivo. To address this question, we have prepared a series of FMRP versions, in which each known in vitro functional domain has been individually deleted, leaving the rest of the protein intact. Constructs with deletions in the protein-protein interaction and RNA-binding as well as in the phosphorylation domains were expressed in STEK-KO cells lacking FMRP and their recruitment into polyribosomal mRNPs and their intra-cellular localization were determined. Our results indicate that the KH RNA-binding domains and the Protein Protein Interacting domain are essential for FMRP to associate with polyribosomal mRNPs, while the RGG box and the phosphorylated domains are dispensable. PMID- 14532326 TI - A mutation in the canine BHD gene is associated with hereditary multifocal renal cystadenocarcinoma and nodular dermatofibrosis in the German Shepherd dog. AB - Hereditary multifocal renal cystadenocarcinoma and nodular dermatofibrosis (RCND) is a naturally occurring canine kidney cancer syndrome that was originally described in German Shepherd dogs. The disease is characterized by bilateral, multifocal tumors in the kidneys, uterine leiomyomas and nodules in the skin consisting of dense collagen fibers. We previously mapped RCND to canine chromosome 5 (CFA5) with a highly significant LOD score of 16.7 (theta=0.016). We have since narrowed the RCND interval following selection and RH mapping of canine genes from the 1.3 x canine genome sequence. These sequences also allowed for the isolation of gene-associated BACs and the characterization of new microsatellite markers. Ordering of newly defined markers and genes with regard to recombinants localizes RCND to a small chromosomal region that overlaps the human Birt-Hogg-Dube locus, suggesting the same gene may be responsible for both the dog and the phenotypically similar human disease. We herein describe a disease-associated mutation in exon 7 of canine BHD that leads to the mutation of a highly conserved amino acid of the encoded protein. The absence of recombinants between the disease locus and the mutation in US and Norwegian dogs separated by several generations is consistent with this mutation being the disease-causing mutation. Strong evidence is provided that the RCND mutation may have a homozygous lethal effect (P<0.01). PMID- 14532327 TI - The usefulness of different density SNP maps for disease association studies of common variants. AB - Large-scale discovery and validation of single-nucleotide polymorphisms (SNPs) facilitates indirect association mapping. It has recently been estimated that, in Europeans, 77% of all SNPs with frequency of 10% or more could be ascertained through linkage disequilibrium (LD) by genotyping variants in the database dbSNP. Using a sampling approach from 73 genes with near complete SNP maps, we show here the usefulness of SNP maps at different densities and the large variability of SNP coverage in different genomic regions. While even sparse SNP maps are of some value to genetic mapping, in order to undertake disease association studies providing at least 80% of SNPs in 90% of genes, much denser maps need to be constructed, at more than one SNP per kb in some regions. PMID- 14532328 TI - Paternal imprints can be established on the maternal Igf2-H19 locus without altering replication timing of DNA. AB - Genomic imprinting in mammals marks the parental alleles in gametes, resulting in differential gene expression in offspring. A number of epigenetic features are associated with imprinted genes. These include differential DNA methylation, histone acetylation and methylation, subnuclear localization and DNA replication timing. While DNA methylation has been shown to be necessary both for establishment and maintenance of imprinting, the connections with the other types of epigenetic marking systems are not clear. Specifically, it is not known whether the other marking systems, either on their own or in conjunction with DNA methylation, are required for imprinting. Here we show that in the mouse mutant Minute (Mnt) the Igf2-H19 locus acquires a paternal methylation imprint in the maternal germline. DNA methylation of the H19 DMR is established in oogenesis, maintained during postzygotic development on the maternal allele, and erased in primordial germ cells. The fact that a paternal type methylation imprint can also be established in the maternal germline indicates that trans-acting factors that target methylation to this imprinted region are likely to be the same in both germlines. Surprisingly, however, asynchrony of DNA replication of the locus is maintained despite the altered expression and methylation imprint of Igf2 and H19. These results show clearly that replication asynchrony of this region is neither the determinant factor for, nor a consequence of, epigenetic modifications that are critical for genomic imprinting. Replication asynchrony may thus be regulated differently from methylation imprints and have a separate function. PMID- 14532329 TI - Noradrenergic neuronal development is impaired by mutation of the proneural HASH 1 gene in congenital central hypoventilation syndrome (Ondine's curse). AB - Congenital central hypoventilation syndrome (CCHS, Ondine's curse) is a rare disorder of the chemical control of breathing. It is frequently associated with a broad spectrum of dysautonomic symptoms, suggesting the involvement of genes widely expressed in the autonomic nervous system. In particular, the HASH-1 PHOX2A-PHOX2B developmental cascade was proposed as a candidate pathway because it controls the development of neurons with a definitive or transient noradrenergic phenotype, upstream from the RET receptor tyrosine kinase and tyrosine hydroxylase. We recently showed that PHOX2B is the major CCHS locus, whose mutation accounts for 60% of cases. We also studied the proneural HASH-1 gene and identified a heterozygous nucleotide substitution in three CCHS patients. To analyze the functional consequences of HASH-1 mutations, we developed an in vitro model of noradrenergic differentiation in neuronal progenitors derived from the mouse vagal neural crest, reproducing in vitro the HASH-PHOX-RET pathway. All HASH-1 mutant alleles impaired noradrenergic neuronal development, when overexpressed from adenoviral constructs. Thus, HASH-1 mutations may contribute to the CCHS phenotype in rare cases, consistent with the view that the abnormal chemical control of breathing observed in CCHS patients is due to the impairment of noradrenergic neurons during early steps of brainstem development. PMID- 14532330 TI - Laforin, the dual-phosphatase responsible for Lafora disease, interacts with R5 (PTG), a regulatory subunit of protein phosphatase-1 that enhances glycogen accumulation. AB - Progressive myoclonus epilepsy of Lafora type (LD, MIM 254780) is a fatal autosomal recessive disorder characterized by the presence of progressive neurological deterioration, myoclonus, epilepsy and polyglucosan intracellular inclusion bodies, called Lafora bodies. Lafora bodies resemble glycogen with reduced branching, suggesting an alteration in glycogen metabolism. Linkage analysis and homozygosity mapping localized EPM2A, a major gene for LD, to chromosome 6q24. EPM2A encodes a protein of 331 amino acids (named laforin) with two domains, a dual-specificity phosphatase domain and a carbohydrate binding domain. Here we show that, in addition, laforin interacts with itself and with the glycogen targeting regulatory subunit R5 of protein phosphatase 1 (PP1). R5 is the human homolog of the murine Protein Targeting to Glycogen, a protein that also acts as a molecular scaffold assembling PP1 with its substrate, glycogen synthase, at the intracellular glycogen particles. The laforin-R5 interaction was confirmed by pull-down and co-localization experiments. Full-length laforin is required for the interaction. However, a minimal central region of R5 (amino acids 116-238), including the binding sites for glycogen and for glycogen synthase, is sufficient to interact with laforin. Point-mutagenesis of the glycogen synthase-binding site completely blocked the interaction with laforin. The majority of the EPM2A missense mutations found in LD patients result in lack of phosphatase activity, absence of binding to glycogen and lack of interaction with R5. Interestingly, we have found that the LD-associated EPM2A missense mutation G240S has no effect on the phosphatase or glycogen binding activities of laforin but disrupts the interaction with R5, suggesting that binding to R5 is critical for the laforin function. These results place laforin in the context of a multiprotein complex associated with intracellular glycogen particles, reinforcing the concept that laforin is involved in the regulation of glycogen metabolism. PMID- 14532331 TI - Haplotype transmission analysis provides evidence of association for DISC1 to schizophrenia and suggests sex-dependent effects. AB - We have previously reported a linkage peak on 1q42 in a Finnish schizophrenia sample. In this study we genotyped 28 single nucleotide polymorphisms (SNPs) from 1q42 covering the three candidate genes TRAX, DISC1 and DISC2, using a study sample of 458 Finnish families ascertained for schizophrenia. Two-point and haplotype association analysis revealed a significant region of interest within the DISC1 gene. A common haplotype (HEP3) was observed to be significantly under transmitted to affected individuals (P=0.0031). HEP3 represents a two SNP haplotype spanning from intron 1 to exon 2 of DISC1. This haplotype also displayed sex differences in transmission distortion, the under-transmission being significant only to affected females (P=0.00024). Three other regions of interest were observed in the TRAX and DISC genes. However, analysis of only those families with complete genotype information specifically highlights the HEP3 haplotype as a true observation. The finding of a common under-transmitted SNP haplotype might imply that this particular allele offers some protection from the development of schizophrenia. Analysis of component-traits of schizophrenia, derived from the Operational Criteria Checklist of Psychotic Illness (OCCPI), displayed association of HEP3 to features of the general phenotype of schizophrenia, including traits representing delusions, hallucinations and negative symptoms. This study provides further evidence for the hypothesis that the DISC1 gene is involved in the aetiology of schizophrenia, and implies a putative sex difference for the effect of the gene. Our findings would also encourage more detailed analyses of the effect of DISC1 on the component-traits of schizophrenia. PMID- 14532332 TI - Hypothalamic malonyl-CoA as a mediator of feeding behavior. AB - Previous studies showed that i.p. administration of C75, a potent inhibitor of fatty acid synthase (FAS), blocked fasting-induced up-regulation of orexigenic neuropeptides and down-regulation of anorexigenic neuropeptides in the hypothalami of mice. As a result, food intake and body weight were drastically reduced. Here we provide evidence supporting the hypothesis that hypothalamic malonyl-CoA, a substrate of FAS, is an indicator of global energy status and mediates the feeding behavior of mice. We use a sensitive recycling assay to quantify malonyl-CoA to show that the hypothalamic malonyl-CoA level is low in fasted mice and rapidly (< or = 2 h) increases (approximately 5-fold) on refeeding. Intracerebroventricular (i.c.v.) administration of C75 to fasted mice rapidly (< or = 2 h) increased (by 4-fold) hypothalamic malonyl-CoA and blocked feeding when the mice were presented with food. Moreover, prior i.c.v. administration of an acetyl-CoA carboxylase inhibitor, 5-(tetradecyloxy)-2-furoic acid, rapidly (although only partially) prevented the C75-induced rise of hypothalamic malonyl-CoA and prevented the C75-induced decrease of food intake. These effects correlated closely with the rapid (< or = 2 h) and reciprocal effects of i.c.v. C75 on the expression of hypothalamic orexigenic (NPY and AgRP) and anorexigenic (proopiomelanocortin) neuropeptide mRNAs. Previous results showing that C75 administered i.c.v. rapidly activates hypothalamic neurons of the arcuate and paraventricular nuclei are consistent with the results reported in this paper. Together these findings suggest that level of hypothalamic malonyl CoA, which depends on the relative activities of acetyl-CoA carboxylase and FAS, is an indicator of energy status and mediates feeding behavior. PMID- 14532333 TI - Microarray experimental design: power and sample size considerations. AB - Gene expression analysis using high-throughput microarray technology has become a powerful approach to study systems biology. The exponential growth in microarray experiments has spawned a number of investigations into the reliability and reproducibility of this type of data. However, the sample size requirements necessary to obtain statistically significant results has not had as much attention. We report here statistical methods for the determination of the sufficient number of subjects necessary to minimize the false discovery rate while maintaining high power to detect differentially expressed genes. Two experimental designs were considered: 1) a comparison between two groups at a single time point, and 2) a comparison of two experimental groups with sequential time points. Computer programs are available for the methods discussed in this paper and are adaptable to more complicated situations. PMID- 14532334 TI - Activation of the protein kinase A pathway in human endometrial stromal cells reveals sequential categorical gene regulation. AB - Decidualization of endometrial stromal cells is a prerequisite for human implantation and occurs in vivo in response to progesterone and involves activation of the protein kinase A (PKA) pathway. The objective of this study was to determine the molecular signatures and patterns of gene expression during stimulation of this pathway with an analog of cAMP. Endometrial stromal cells from two subjects were treated with or without 8-Br-cAMP (1 mM) for 0, 2, 12, 24, 36, and 48 h and were processed for microarray analysis, screening for 12,686 genes and ESTs. Most abundantly upregulated genes included neuropeptides, immune genes, IGF family members, cell cycle regulators, extracellular matrix proteases, cholesterol trafficking, cell growth and differentiation, hormone signaling, and signal transduction. Most abundantly downregulated genes included activator of NF kappaB, actin/tropomyosin/calmodulin binding protein, cyclin B, IGFBP-5, alpha1 type XVI collagen, lipocortin III, l-kynurenine hydrolase, frizzle-related protein, and cyclin E2. RT-PCR validated upregulation of IGFBP-1, preprosomatostatin, and IL-11, and Northern analysis validated their kinetic upregulation. RT-PCR confirmed downregulation of IGFBP-5, cyclin B, and TIL-4. K means analysis revealed four major patterns of up- and downregulated genes, and genes within each ontological group were categorized into these four kinetic patterns. Within each ontological group different patterns of temporal gene expression were observed, indicating that even genes within one functional category are regulated differently during activation of the PKA pathway in human endometrial stromal cells. Overall, the data demonstrate kinetic reprogramming of genes within specific functional groups and changes in genes associated with nucleic acid binding, cell proliferation, decreased G protein signaling, increased STAT pathway signaling, structural proteins, cellular differentiation, and secretory processes. These changes are consistent with cAMP modulating early events (0-6 h) primarily involving cell cycle regulation, subsequent events (12 24 h) involving cellular differentiation (including changes in morphology and secretory phenotype), and late events (24-48 h) mediating more specialized function, including immune modulators, in the human endometrial stromal cell. PMID- 14532335 TI - Genomic map of cardiovascular phenotypes of hypertension in female Dahl S rats. AB - Genetic linkage analyses in human populations have traditionally combined male and female progeny for determination of quantitative trait loci (QTL). In contrast, most rodent studies have focused primarily on males. This study represents an extensive female-specific linkage analysis in which 236 neuroendocrine, renal, and cardiovascular traits related to arterial pressure (BP) were determined in 99 female F2 rats derived from a cross of Dahl salt sensitive SS/JrHsdMcwi (SS) and Brown Norway normotensive BN/SsNHsdMcwi (BN) rats. We identified 126 QTL for 96 traits on 19 of the 20 autosomal chromosomes of the female progeny. Four chromosomes (3, 6, 7, and 11) were identified as especially important in regulation of arterial pressure and renal function, since aggregates of 8-11 QTL mapped together on these chromosomes. BP QTL in this female population differed considerably from those previously found in male, other female, or mixed sex population linkage analysis studies using SS rats. Kidney weight divided by body weight was identified as an intermediate phenotype that mapped to the same region of the genome as resting diastolic blood pressure and was correlated with that same BP phenotype. Seven other phenotypes were considered as "potential intermediate phenotypes, " which mapped to the same region of the genome as a BP QTL but were not correlated with BP. These included renal vascular responses to ANG II and ACh and indices of baroreceptor responsiveness. Secondary traits were also identified that were likely to be consequences of hypertension (correlated with BP but not mapped to a BP QTL). Seven such traits were found, notably heart rate, plasma cholesterol, and renal glomerular injury. The development of a female rat systems biology map of cardiovascular function represents the first attempt to prioritize those regions of the genome important for development of hypertension and end organ damage in female rats. PMID- 14532336 TI - Mutation in tcpR gene (Vc0832) of Vibrio cholerae O1 causes loss of tolerance to high osmolarity and affects colonization and virulence in infant mice. AB - Vibrio cholerae, the agent of cholera, multiplies and colonizes human intestinal tract where it survives high osmolarity due to bile and other sodium salts. In this work, by TnphoA mutagenesis, a mutant of V. cholerae O1 which could not grow and form colonies on LB agar containing 400 mM NaCl has been characterized. The mutant, designated CD83, adhered normally to freshly isolated rabbit intestinal discs, colonized poorly the gut of infant mice and was avirulent in the same model, whereas the parent strain CD81 would colonize the gut and cause death of mice. Attenuation of virulence of CD83 was not attributable to its inability to produce cholera toxin, as no difference was found in the cholera toxin produced in vitro by the mutant and parent strains. Molecular cloning and sequencing of the mutated gene revealed that insertion of transposon occurred in tcpR gene (Vc0832) of V. cholerae. Complementation of the mutant with wild-type tcpR gene resulted in restoration of the ability to survive at high salt concentration (400 mM NaCl), and to colonize the gut and restore virulence. The results suggest that tcpR plays a role in survival of V. cholerae in the small intestine of host as the osmolarity in the intestinal lumen is thought to be equivalent to 300 mM NaCl or higher. PMID- 14532337 TI - Subtyping of virulence genes in verocytotoxin-producing Escherichia coli (VTEC) other than serogroup O157 associated with disease in the United Kingdom. AB - Verocytotoxin-producing Escherichia coli (VTEC) causes a wide spectrum of disease in humans, from mild diarrhoea to haemolytic uraemic syndrome (HUS). The verocytotoxin (vtx) and intimin (eae) genes of VTEC strains, other than those of serogroup O157, were subtyped to identify common properties that may be associated with increased pathogenicity. Strains were isolated from patients with HUS, those with diarrhoea or from asymptomatic individuals. Strains of VTEC that carried vtx(2) gene subtypes vtx(2) and vtx(2c) were most commonly associated with HUS, whereas strains from patients with less severe disease and from the healthy control group were more likely to have vtx(1c) or vtx(2d) genes. The eae gene was detected more frequently in strains isolated from HUS patients than in those associated with cases of diarrhoea; beta-intimin was the most common intimin subtype in strains isolated from both groups of patients. None of the strains from the healthy control group carried the eae gene. PMID- 14532338 TI - Increased prevalence of seropositivity for non-gastric Helicobacter species in patients with autoimmune liver disease. AB - Various Helicobacter species have been isolated from the stomach, intestinal tract and liver of a variety of mammalian and some avian species, and Helicobacter DNA has been detected in human bile and liver samples. An immunoblot assay was established to analyse serum antibody responses to non-gastric Helicobacter species in patients with autoimmune liver diseases, in comparison with healthy individuals. Sera from 36 patients with primary sclerosing cholangitis (PSC), 21 with primary biliary cirrhosis, 19 with autoimmune chronic hepatitis and 80 blood donors were analysed by immunoblot, using cell-surface proteins from Helicobacter pullorum, Helicobacter bilis and Helicobacter hepaticus as antigens. Prior to testing, sera were cross-absorbed with a whole cell lysate of Helicobacter pylori. Antibody reactivity to various proteins of these three Helicobacter species was measured by densitometric scanning and results were processed by computer software to estimate antigenic specificity. Results were also compared with antibody response to H. pylori. For H. pullorum, reactivity to at least two of the proteins with molecular masses of 48, 45, 37, 20 and 16 kDa, for H. hepaticus, reactivity to the 76, 30 and 21 kDa proteins and for H. bilis, reactivity to the 22 and 20 kDa proteins, seemed to have high specificity. Positive immunoblot results with sera from patients with PSC to antigens of H. pullorum, H. bilis and H. hepaticus were found in 38, 22 and 25 % of cases, respectively, and from patients with other autoimmune liver diseases, in 30, 22 and 22 % of cases, respectively. Prevalence of serum antibodies to non gastric Helicobacter species was significantly higher in patients with autoimmune chronic liver diseases than in healthy blood donors (P < 0.001). Increased antibody levels to enterohepatic Helicobacter species raise questions concerning an infectious role of these emerging bacterial pathogens in human autoimmune liver diseases. PMID- 14532339 TI - Characterization of variants of the gene encoding the p55 antigen in Pneumocystis from rats and mice. AB - Variants of the p55 gene in rat-derived Pneumocystis carinii have been identified and its counterpart in mouse-derived P. carinii f. sp. muris has been cloned. By PCR amplification of P. carinii genomic DNA, five variants were identified that differed from each other in size and sequence, primarily in the number and size of encoded amino acid repeats. For P. carinii f. sp. muris, a single PCR fragment (471 bp) was obtained, which contained an incomplete ORF encoding a 157 aa protein that was most similar to a p55 variant in P. carinii, with nucleotide and amino acid sequence identity of 79 and 68 %, respectively. Southern blot analysis revealed the presence of more than one copy of the p55 gene in both Pneumocystis species. Thus, like other Pneumocystis antigens, p55 exhibits polymorphism that could potentially benefit the organism in host interactions. PMID- 14532340 TI - Cryptococcus neoformans induces alterations in the cytoskeleton of human brain microvascular endothelial cells. AB - The fungal pathogen Cryptococcus neoformans has a predilection for the central nervous system (CNS), resulting in devastating meningoencephalitis. At present, it is unclear how C. neoformans traverses the blood-brain barrier (BBB) and causes CNS infection. The present study has examined and characterized the interaction of C. neoformans with human brain microvascular endothelial cells (HBMEC), which constitute the BBB. Adhesion of and transcytosis of HBMEC by C. neoformans was inoculum- and time-dependent and occurred with both encapsulated and acapsulated strains. C. neoformans induced marked morphological changes in HBMEC, for example membrane ruffling, irregular nuclear morphology and swelling of the mitochondria and the ER. These findings suggest that C. neoformans induced actin cytoskeletal reorganization of the host cells. In addition, it was observed that the dephosphorylated form of cofilin was increased during cryptococcal adherence to HBMEC, concomitant with the actin rearrangement. Cryptococcal binding to HBMEC was increased in the presence of Y27632, a Rho kinase (ROCK) specific inhibitor. Since ROCK activates LIM kinase (LIMK), which phosphorylates cofilin (inactive form), this suggests the involvement of the ROCK-->LIMK- >cofilin pathway. In contrast, the phosphatase inhibitor sodium orthovanadate decreased adherence of Cryptococcus to HBMEC, concomitant with the increase of phosphorylation of cofilin. Furthermore, the tight junction marker protein occludin became Triton-extractable, indicating alteration of tight junctions in brain endothelial cells. This is the first demonstration that C. neoformans is able to adhere to and transcytose across the HBMEC monolayer and alter the cytoskeleton morphology in HBMEC. Further characterization of the interactions between C. neoformans and HBMEC should help the development of novel strategies to prevent cryptococcal meningitis and its associated morbidity. PMID- 14532341 TI - High levels of hydrolytic enzymes secreted by Candida albicans isolates involved in respiratory infections. AB - Differences in production of two putative virulence factors of Candida albicans, phospholipase and proteinase, were determined for a large panel of clinical C. albicans isolates (n = 186) obtained from the European SENTRY programme. Seventy two per cent of isolates produced detectable amounts of phospholipase and 95 % of isolates produced detectable amounts of proteinase. There was no clear correlation between the results of the phospholipase and proteinase assays and the geographical distribution of the isolates. However, isolates that originated from respiratory infections produced significantly higher amounts of phospholipase than isolates obtained from blood, the urinary tract or wounds/skin/soft tissue and also appeared to produce more proteinase. These virulent isolates involved in respiratory infections may originate from the oral cavity. Whether these results are caused by selection for these highly virulent isolates remains to be resolved. PMID- 14532342 TI - Evaluation of a new serotyping kit for Streptococcus pneumoniae. AB - A new serotyping test kit (Streptococcus pneumoniae antisera "Seiken" set; Denka kit) was evaluated for 285 strains of Streptococcus pneumoniae in comparison with the standard capsular reaction (Quellung test). This new kit is based on the slide-agglutination method and is composed of eight pool sera, 40 group or type sera and 41 specific type sera. All serotyping results by using the Denka kit were completely identical to those obtained by using the conventional Quellung test. For types and groups, sensitivity and specificity were 100 and 100 %, respectively. For specific types, sensitivity and specificity were 100 and 100 %, respectively. The Denka kit is relatively rapid (mean test time, 5 min, versus 15 min by Quellung test), cheap (0.5 US$ per test, versus 1.4 US$ per Quellung test), easy to perform and does not require special equipment. The Denka kit may be useful for fieldworkers in developing countries involved in epidemiological surveys and vaccine development. PMID- 14532343 TI - Characterization of German penicillin non-susceptible serotype 23F pneumococci using multilocus sequence typing. AB - Three nationwide multicentre studies (n = 5071) showed an increase in antibiotic resistance in pneumococci in Germany. Serotype 23F was the predominant serotype (n = 45, 22.4 %), followed by 6B (n = 30, 14.9 %) and 9V (n = 19, 9.5 %). Multilocus sequence typing was used to characterize 45 serotype 23F strains with reduced penicillin susceptibility. The Spanish(23F)-1 clone [profile 4-4-2-4-4-1 1, sequence type (ST) 81] contributes significantly to the emergence of penicillin resistance in Germany (n = 21, 46.7 % of all penicillin non susceptible serotype 23F isolates). Isolates of ST 277 (profile 7-13-8-6-6-12-8), which has been found previously in the Netherlands, are also observed, particularly in western Germany (n = 8, 17.8 %). A high proportion of strains (n = 11, 24.4 %) have sequence types that have not been reported to date from other countries (STs 353-362). The major penicillin-resistant clones are present in Germany, a country with relatively low levels of beta-lactam resistance. PMID- 14532344 TI - Evaluation of a multilocus sequence typing system for Staphylococcus epidermidis. AB - Staphylococcus epidermidis is a significant cause of nosocomial disease. However, the taxonomy of this pathogen, particularly at subspecies level, is unclear. A multilocus sequence typing (MLST) scheme has therefore been investigated as a tool to elucidate taxonomic relationships within this group, based on genetic relatedness. DNA sequences for internal fragments of seven housekeeping genes were compared in 47 geographically and temporally diverse S. epidermidis isolates that were obtained from clinical infections. Twenty-three different allelic profiles were detected; 17 of these were represented by single strains and the largest profile group contained 17 isolates. Diversity of the same collection of isolates was investigated by using PFGE of SmaI-digested genomic DNA to test the discrimination and validity of the MLST approach. Isolates within the largest profile group were resolved into four distinct PFGE clusters on the basis of their SmaI digest patterns. Isolates within other profile groups that contained multiple isolates had matching PFGE SmaI patterns within each group. It appears that MLST is an effective method for grouping S. epidermidis strains at the subspecies level; however, it is not as discriminatory as it has been for other species for which MLST schemes have been established and, used alone, would not be a useful method for epidemiological studies. In addition, it was demonstrated that this method was effective for confirming the identity of S. epidermidis CoNS (coagulase-negative) isolates. PMID- 14532345 TI - Detection of katG Ser315Thr substitution in respiratory specimens from patients with isoniazid-resistant Mycobacterium tuberculosis using PCR-RFLP. AB - Mutations in the katG locus of catalase peroxidase in Mycobacterium tuberculosis (MTB) account for major isoniazid (INH) resistance. In the South China region, a collection of 906 respiratory specimens and 142 MTB isolates was used to evaluate the sensitivity and specificity of a PCR-RFLP method for the detection of INH resistance-associated mutations. Except for four catalase-negative MTB isolates, katG PCR for a 620-bp amplicon was successful for all purified MTB isolates. For respiratory specimens, diagnostic sensitivity and specificity of katG PCR was 85 and 100 %. Subsequent RFLP of the katG amplicons by MspI digestion identified that 51 % of INH-resistant MTB were associated with the Thr315 phenotype, and that codon 463 was a polymorphic site with no linkage to INH resistance. The Arg463 wild-type MTB isolates predominant in the Western world were replaced by isolates carrying Leu463 in the South China region. RFLP patterns of katG amplicons from respiratory specimens were identical to those of the corresponding MTB cultured colonies. This method has potential application for rapid diagnosis of INH resistance due to katG Ser315Thr mutation. PMID- 14532346 TI - Comparison of usefulness of randomly amplified polymorphic DNA and amplified fragment length polymorphism techniques in epidemiological studies on nasopharyngeal carriage of non-typable Haemophilus influenzae. AB - Randomly amplified polymorphic DNA (RAPD) and automated amplified-fragment length polymorphism (AFLP) techniques with fluorescently labelled primers were used to type non-serotypable Haemophilus influenzae (NTHI) isolates. Eighty-seven isolates from healthy children attending day-care centres or living at orphanages in southern Poland were investigated. Through comparison of the AFLP data with RAPD analysis, it has been concluded that the discriminatory power of AFLP for NTHI typing is higher than RAPD. Generally, the NTHI isolates analysed were highly heterogeneous, as detected with a HindIII/TaqI AFLP genotyping scheme on intra/inter similarity levels of 94 and 96 % using Pearson's correlation coefficient. The range of similarity values found for isolates from children permanently residing at a particular day-care centre was much wider than that for isolates from orphanages. AFLP can efficiently access NTHI strain diversity and can monitor their turn-over for comparative typing in local and inter-local epidemiological investigations. PMID- 14532347 TI - High prevalence of atypical enteropathogenic Escherichia coli (EPEC) in Norwegian children with diarrhoea. AB - The aim of the present study was to investigate the relative contribution of enteropathogenic Escherichia coli (EPEC) as a cause of infectious diarrhoea in Norwegian children. Data from faecal specimens from children <2 years old with diarrhoea during the year 2001 were analysed. E. coli isolates with the attaching and effacing genotype (eae+) were examined for the presence of the bundle-forming pilus (bfpA) and Shiga toxin genes by PCR, and for genetic relatedness by PFGE. During the 1-year period, 598 specimens from 440 patients <2 years old were analysed. Potential enteric pathogens were identified in 124 patients (28.2 %). EPEC was the most frequently identified agent (44 patients), followed by rotavirus (41 patients), Campylobacter jejuni (17 patients) and adenovirus (17 patients). All other agents were detected in five patients or less. Only one of the eae+ E. coli isolates was classified as typical EPEC (bfpA+). Among the 43 isolates that were classified as atypical EPEC (bfpA-), eight strains belonged to EPEC serogroups, whereas the majority of strains (n = 35) were not agglutinated by EPEC antisera. None of the EPEC isolates were genetically related. This study demonstrates that atypical EPEC of non-EPEC serogroups is highly prevalent among Norwegian children with diarrhoea. PMID- 14532348 TI - Experimental Ehrlichia chaffeensis infection in beagles. AB - A canine model for human monocytic ehrlichiosis was used to assess persistent infection and antigenic variation of Ehrlichia chaffeensis. Two beagle dogs were infected subcutaneously with E. chaffeensis Arkansas strain. The dogs were observed for 6 months after inoculation for clinical signs, blood chemistry changes, antibodies to E. chaffeensis and presence of E. chaffeensis in the blood. Both dogs developed thrombocytopenia, but exhibited normal body temperatures during the entire course of infection. In one dog, E. chaffeensis was cultivated for up to 74 days post-inoculation and E. chaffeensis DNA was detected in the dog's blood for up to 81 days. In the other dog, E. chaffeensis was cultured for up to 102 days and E. chaffeensis DNA was detected in the blood for up to 117 days. PCR amplification and DNA sequence analysis indicated that there was no genetic variation in the 120 kDa outer-membrane glycoprotein gene of E. chaffeensis during infection of the dogs. The dogs developed antibodies to the immunodominant proteins of E. chaffeensis, including the 175, 140, 120, 80, 50 and 28 kDa proteins, starting in the fifth week post-inoculation. The dogs maintained high antibody titres throughout the 6-month study period. These results indicate that dogs become carriers of E. chaffeensis for 2-4 months after infection without exhibiting signs of clinical disease, suggesting that dogs may serve as a natural host for E. chaffeensis. PMID- 14532349 TI - Absence of Mycoplasma species DNA in chronic fatigue syndrome. PMID- 14532350 TI - Susceptibility to antimicrobials and mechanisms of erythromycin resistance in clinical isolates of Streptococcus agalactiae from Rio de Janeiro, Brazil. PMID- 14532351 TI - Clinical insignificance of (1-->3)-beta-D-glucan in early diagnosis of invasive pulmonary aspergillosis in a patient with chronic obstructive pulmonary disease. PMID- 14532352 TI - Comprehensive proteomic profiling of the membrane constituents of a Mycobacterium tuberculosis strain. AB - Mycobacterium tuberculosis is an infectious microorganism that causes human tuberculosis. The cell membranes of pathogens are known to be rich in possible diagnostic and therapeutic protein targets. To compliment the M. tuberculosis genome, we have profiled the membrane protein fraction of the M. tuberculosis H37Rv strain using an analytical platform that couples one-dimensional SDS gels to a microcapillary liquid chromatography-nanospray-tandem mass spectrometer. As a result, 739 proteins have been identified by two or more distinct peptide sequences and have been characterized. Interestingly, approximately 450 proteins represent novel identifications, 79 of which are membrane proteins and more than 100 of which are membrane-associated proteins. The physicochemical properties of the identified proteins were studied in detail, and then biological functions were obtained by sorting them according to Sanger Institute gene function category. Many membrane proteins were found to be involved in the cell envelope, and those proteins with energy metabolic functions were also identified in this study. PMID- 14532353 TI - Profiling core proteomes of human cell lines by one-dimensional PAGE and liquid chromatography-tandem mass spectrometry. AB - Protein expression profiles vary considerably between human cell lines and tissues, which is in part a reflection of their specialized roles within an organism. It is of considerable practical use to establish which proteins constitute the primary components of the respective proteomes. When compiled into databases, such information can facilitate the assessment of selectivity and specificity of a wide range of proteomic experiments. Here we describe the major constituents of proteomes of six human immortalized cell lines. By employing a combination of one-dimensional SDS-PAGE and nanocapillary liquid chromatography tandem mass spectrometry (LC-MS/MS), we identified up to 1785 non-redundant cytoplasmic and nuclear proteins from a single cell line using 50 and 30 microg of total protein from the corresponding fractions. Up to 38 proteins could be identified from a single band in one liquid chromatography-MS/MS experiment. When combined with systematic gridding of gel lanes into 48 slices, a dynamic range for protein identification of approximately 1:2000 can be envisaged for this approach. Identified proteins range from 4-553 kDa in size, cover the pI range between 3.4 and 12.8, and include 255 proteins with predicted transmembrane domains. Repeated analysis of peptides derived from the same gel band showed that the reproducibility of nanocapillary liquid chromatography-MS/MS of such complex mixtures is about 60-70% suggesting that a particular analytical experiment would need to be repeated about three times to arrive at a representative estimate of the set of highly abundant proteins in a given proteome. Given its technical simplicity, sensitivity, and wealth of generated information, we have adopted this experimental approach to characterize every cell line and tissue that is the subject of experimentation in our laboratory. The combined dataset for the six cell lines consists of 2341 non-redundant human proteins and thus constitutes one of the largest collections of human proteomic data published to date. PMID- 14532355 TI - Signal transduction underlying carbachol-induced contraction of rat urinary bladder. II. Protein kinases. AB - We have investigated the role of several protein kinases in carbachol-stimulated, M(3) muscarinic receptor-mediated contraction of rat urinary bladder. Concentration-response curves for the muscarinic receptor agonist carbachol were generated in the presence of multiple concentrations of inhibitors of various protein kinases, their inactive controls, or their vehicles. Bladder contraction was not significantly inhibited by three protein kinase C inhibitors (chelerythrine, 1-10 microM; calphostin C, 0.1-1 microM; and 2-[1-(3 dimethylaminopropyl)-1H-indol-3-yl]-3-(1H-indol-3-yl)-maleimide (Go 6850), 1-10 microM), by the tyrosine kinase inhibitor genistein or its inactive control daidzein (3-30 microM each), or by two inhibitors of activation of mitogen activated protein kinase [10-100 microM 2'-amino-3'-methoxyflavone (PD 98,059) and 3-30 microM 1,4-diamino-2,3-dicyano-1,4-bis(2-aminophenylthio)butadiene (U 124)] or their negative control 1,4-diamino-2,3-dicyano-1,4 bis(methylthio)butadiene (U 126) (3-30 microM). Although high concentrations of wortmannin (3-30 microM) inhibited bladder contraction, this was not mimicked by another inhibitor of phosphatidylinositol-3-kinase, 2-(4-morpholinyl)-8-phenyl-4H 1-benzopyran-4-one (LY 294,002) (3-30 microM) and, hence, was more likely due to direct inhibition of myosin light chain kinase by wortmannin than to an involvement of phosphatidylinositol-3-kinase. In contrast, trans-4-[(1R)-1 aminoethyl]-N-4-pyridinylcyclohexanecarboxamide (Y 27,632) (1-10 microM), an inhibitor of rho-associated kinases, concentration-dependently and effectively attenuated the carbachol responses. We conclude that carbachol-induced contraction of rat urinary bladder does not involve protein kinase C, phosphatidylinositol-3-kinase, tyrosine kinases, or extracellular signal regulated kinases; in contrast, rho-associated kinases appear to play an important role in the regulation of bladder contraction. PMID- 14532354 TI - Signal transduction underlying carbachol-induced contraction of rat urinary bladder. I. Phospholipases and Ca2+ sources. AB - We have reexamined the muscarinic receptor subtype mediating carbachol-induced contraction of rat urinary bladder and investigated the role of phospholipase (PL)C, D, and A2 and of intra- and extracellular Ca2+ sources in this effect. Based on the nonsubtype-selective tolterodine, the highly M2 receptor-selective (R)-4-[2-[3-(4-methoxy-benzoylamino)-benzyl]-piperidin-1-ylmethyl]-piperidine-1 carboxylic acid amide (Ro-320-6206), and the highly M3 receptor-selective darifenacin and 3-(1-carbamoyl-1,1-diphenylmethyl)-1-(4 methoxyphenylethyl)pyrrolidine (APP), contraction occurs via M3 receptors. Carbachol stimulated inositol phosphate formation in rat bladder slices, and this was abolished by the phospholipase C inhibitor 1-(6-[([17beta]-3-methoxyestra 1,3,5[10]-trien-17-yl)-amino]hexyl)-1H-pyrrole-2,5-dione (U 73,122; 10 microM). Nevertheless, U 73,122 (1-10 microM) did not significantly affect carbachol stimulated bladder contraction. Carbachol had only little effect on PLD activity in bladder slices, but the PLD inhibitor butan-1-ol, relative to its negative control butan-2-ol (0.3% each), caused detectable inhibition of carbachol-induced bladder contraction. The cytosolic PLA2 inhibitor arachidonyltrifluoromethyl ketone weakly inhibited carbachol-induced contraction at a concentration of 300 microM, but the cyclooxygenase inhibitor indomethacin (1-10 microM) remained without effect. The Ca2+ entry blocker nifedipine (10-100 nM) almost completely inhibited carbachol-induced bladder contraction. In contrast, 1-[beta-[3-(4 methoxyphenyl)propoxy]-4-methoxyphenethyl]-1H-imidazole HCl (SKF 96,365; 10 microM), an inhibitor of store-operated Ca2+ channels, caused little inhibition. We conclude that carbachol-induced contraction of rat bladder largely depends on Ca2+ entry through nifedipine-sensitive channels and, perhaps, PLD, PLA2, and store-operated Ca2+ channels, whereas cyclooxygenase and, surprisingly, also PLC are not involved to a relevant extent. PMID- 14532356 TI - A tragic death: a time to blame or a time to learn? PMID- 14532357 TI - Using research knowledge to improve health care. PMID- 14532358 TI - Medical school differences: beneficial diversity or harmful deviations? PMID- 14532359 TI - Monitoring clinical outcomes in primary care. PMID- 14532360 TI - What can ethnography do for quality and safety in health care? PMID- 14532361 TI - Basing decisions on better quality routine clinical data. PMID- 14532362 TI - Deriving recommendations in clinical practice guidelines. PMID- 14532363 TI - Medical school attended as a predictor of medical malpractice claims. AB - OBJECTIVES: Following earlier research which showed that certain types of physicians are more likely to be sued for malpractice, this study explored (1). whether graduates of certain medical schools have consistently higher rates of lawsuits against them, (2). if the rates of lawsuits against physicians are associated with their school of graduation, and (3). whether the characteristics of the medical school explain any differences found. DESIGN: Retrospective analysis of malpractice claims data from three states merged with physician data from the AMA Masterfile (n=30288). STUDY SUBJECTS: All US medical schools with at least 5% of graduates practising in three study states (n=89). MAIN OUTCOME MEASURES: Proportion of graduates from a medical school for a particular decade sued for medical malpractice between 1990 and 1997 and odds ratio for lawsuits against physicians from high and low outlier schools; correlations between the lawsuit rates of successive cohorts of graduates of specific medical schools. RESULTS: Medical schools that are outliers for malpractice lawsuits against their graduates in one decade are likely to retain their outlier status in the subsequent decade. In addition, outlier status of a physician's medical school in the decade before his or her graduation is predictive of that physician's malpractice claims experience (p<0.01). All correlations of cohorts were relatively high and all were statistically significant at p<0.001. Comparison of outlier and non-outlier schools showed that some differences exist in school ownership (p<0.05), years since established (p<0.05), and mean number of residents and fellows (p<0.01). CONCLUSIONS: Consistent differences in malpractice experience exist among medical schools. Further research exploring alternative explanations for these differences needs to be conducted. PMID- 14532364 TI - Is it possible and worth keeping track of deaths within general practice? Results of a 15 year observational study. AB - AIM: To assess the value of maintaining a death register in a general practice with particular reference to monitoring quality of care. DESIGN OF STUDY: Observational study. SETTING: Inner London general practice. METHOD: The practice maintained a manual death register, retained medical records of all deceased patients, and requested information on cause of death from health authorities and coroners for 15 years. MAIN OUTCOME MEASURES: Number and causes of deaths; 3 yearly age standardised death rates; proportion of deaths formally notified to the practice; place of death; source of cause of death information. RESULTS: During the study period 578 patients died. Practice age standardised death rates fell significantly from 35.59 to 27.12/1000. 498 (86.2%) deaths were formally notified to the practice, 392 within 7 days of death. Of 143 deaths reported to the coroner, only 45 coroners' reports were received. 360 (64.1%) died in hospital, 139 (24.8%) at home, and 38 (6.8%) in a hospice. Death certificate cause of death information was obtained from patients' records in 33.6% (n=194) of cases and from health authority sources for 50% (n=289). The pattern of ascertained causes of deaths was similar to the national pattern. CONCLUSION: A death register can examine trends in practice deaths by age and place of death and comparisons undertaken with nationally published mortality data. An accurate picture of cause of death cannot be generated from routine data flows alone. There is delay in informing GPs of patient deaths. Meaningful and timely monitoring of deaths cannot be undertaken by individual practices. National Statistics should provide routine analysis of GP death certificate information. PMID- 14532365 TI - Causes of intravenous medication errors: an ethnographic study. AB - BACKGROUND: Intravenous (IV) medication errors are frequent events. They are associated with considerable harm, but little is known about their causes. Human error theory is increasingly used to understand adverse events in medicine, but has not yet been applied to study IV errors. Our aim was to investigate causes of errors in IV drug preparation and administration using a framework of human error theory. METHODS: A trained and experienced observer accompanied nurses during IV drug rounds on 10 wards in two hospitals (one university teaching hospital and one non-teaching hospital) in the UK. Information came from observation and talking informally to staff. Human error theory was used to analyse the causes of IV error. RESULTS: 265 IV drug errors were identified during observation of 483 drug preparations and 447 administrations. The most common type of error was the deliberate violation of guidelines when injecting bolus doses faster than the recommended speed of 3-5 minutes. Causes included a lack of perceived risk, poor role models, and available technology. Mistakes occurred when drug preparation or administration involved uncommon procedures such as the preparation of very small volumes or the use of unusual drug vial presentations. Causes included a lack of knowledge of preparation or administration procedures and complex design of equipment. Underlying problems were the cultural context allowing unsafe drug use, the failure to teach practical aspects of drug handling, and design failures. CONCLUSIONS: Training needs and design issues should be addressed to reduce the rate of IV drug preparation and administration errors. This needs a coordinated approach from practitioners, regulators, and the pharmaceutical industry. PMID- 14532366 TI - Directory of clinical databases: improving and promoting their use. AB - BACKGROUND: The controversy surrounding the actual and potential use of clinical databases partly reflects the huge variation in their content and quality. In addition, use of existing clinical databases is severely limited by a lack of knowledge of their availability. OBJECTIVES: To develop and test a standardised method for assessing the quality (completeness and accuracy) of clinical databases and to establish a web based directory of databases in the UK. METHODS: An expert group was set up (1). to establish the criteria for inclusion of databases; (2). to develop a quality assessment instrument with high content validity, based on epidemiological theory; (3). to test empirically, modify, and retest the acceptability to database custodians, face validity and floor/ceiling effects; and (4). to design a website. RESULTS: Criteria for inclusion of databases were the provision of individual level data; inclusion in the database defined by a common circumstance (e.g. condition, treatment), an administrative arrangement, or an adverse outcome; and inclusion of data from more than one provider. A quality assessment instrument consisting of 10 items (four on coverage, six on reliability and validity) was developed and shown to have good face and content validity, no floor/ceiling effects, and to be acceptable to database custodians. A website (www.docdat.org) was developed. Indications over the first 18 months (number of visitors to the site) are that it is increasingly popular. By November 2002 there were around 3500 hits a month. CONCLUSIONS: A website now exists where visitors can identify clinical databases in the UK that may be suitable to meet their aims. It is planned both to develop a local version for use within a hospital and to encourage similar national systems in other countries. PMID- 14532368 TI - Make no mistake-errors can be controlled. AB - Traditional quality control methods identify "variation" as the enemy. However, the control of variation by itself can never achieve the remarkably low non conformance rates of world class quality leaders. Because the control of variation does not achieve the highest levels of quality, an inordinate focus on these techniques obscures key quality improvement opportunities and results in unnecessary pain and suffering for patients, and embarrassment, litigation, and loss of revenue for healthcare providers. Recent experience has shown that mistakes are the most common cause of problems in health care as well as in other industrial environments. Excessive product and process complexity contributes to both excessive variation and unnecessary mistakes. The best methods for controlling variation, mistakes, and complexity are each a form of mistake proofing. Using these mistake proofing techniques, virtually every mistake and non-conformance can be controlled at a fraction of the cost of traditional quality control methods. PMID- 14532369 TI - Value for money of changing healthcare services? Economic evaluation of quality improvement. AB - There are many instances of perceived or real inefficiencies in health service delivery. Both healthcare providers and policy makers need to know the impact and cost of applying strategies to change the behaviour of individuals or organisations. Quality improvement or implementation research is concerned with evaluating the methods of behavioural change. Addressing inefficiencies in healthcare services raises a series of issues, beginning with how inefficiency itself should be defined. The basic concepts of cost analysis and economic evaluations are explained and a model for working through the economic issues of quality improvement is discussed. This model combines the costs and benefits of corrected inefficiency with the costs and degree of behavioural change achieved by a quality improvement method in the policy maker's locality. It shows why it may not always be cost effective for policy makers to address suboptimal behaviour. Both the interpretation of quality improvement research findings and their local application need careful consideration. The limited availability of applicable quality improvement research may make it difficult to provide robust advice on the value for money of many behavioural quality improvement strategies. PMID- 14532367 TI - Ethnic specific recommendations in clinical practice guidelines: a first exploratory comparison between guidelines from the USA, Canada, the UK, and the Netherlands. AB - OBJECTIVES: To investigate whether clinical practice guidelines in different countries take ethnic differences between patients into consideration and to assess the scientific foundation of such ethnic specific recommendations. DESIGN: Analysis of the primary care sections of clinical practice guidelines. SETTING: Primary care practice guidelines for type 2 diabetes mellitus, hypertension, and asthma developed in the USA, Canada, the UK, and the Netherlands. MAIN OUTCOME MEASURES: Enumeration of the ethnic specific information and recommendations in the guidelines, and the scientific basis and strength of this evidence. RESULTS: Different guidelines do address ethnic differences between patients, but to a varying extent. The USA guidelines contained the most ethnic specific statements and the Dutch guidelines the least. Most ethnic specific statements were backed by scientific evidence, usually arising from descriptive studies or narrative reviews. CONCLUSION: The attention given to ethnic differences between patients in clinical guidelines varies between countries. Guideline developers should be aware of the potential problems of ignoring differences in ethnicity. PMID- 14532370 TI - Promoting quality improvement in French healthcare organisations: design and impact of a compendium of models and tools. AB - Relevant and user friendly information should be provided to professionals who wish to promote quality improvement in healthcare organisations (HCOs). In response to requests from French HCOs, we designed a compendium of methods and tools for use in quality improvement. Its contents were based on a critical review of the literature, face-to-face interviews with three industrial/business experts in quality, the views of 13 healthcare professionals knowledgeable in quality issues, and comments from over 40 potential users of the compendium. Overall, 14 methods and 20 tools relevant and applicable to the healthcare sector were identified. They were classified according to their main thrust, explained in detail, illustrated with specific cases from the literature or from personal experience, and published as a loose leaf compendium. The compendium was posted on the worldwide web and presented to healthcare managers in September 2000. It has become one of the most popular ANAES publications (approximately 5400 downloads over the first 6 months), partly because all French HCOs are legally bound to undergo accreditation which has been set up and is being implemented by ANAES. PMID- 14532371 TI - The role of error in organizing behaviour. 1990. AB - During recent years the significance of the concept of human error has changed considerably. The reason for this has partly been an increasing interest of psychological research in the analysis of complex real life phenomena, and partly the changes of modern work conditions caused by advanced information technology. Consequently, the topic of the present contribution is not a definition of the concept or a proper taxonomy. Instead, a review is given of two professional contexts for which the concept of error is important. Three cases of analysis of human-system interaction are reviewed: (1). traditional task analysis and human reliability estimation; (2). causal analysis of accidents after the fact, and (3). design of reliable work conditions in modern sociotechnical systems. It is concluded that "errors" cannot be studied as a separate category of behaviour fragments; the object of study should be cognitive control of behaviour in complex environments. PMID- 14532374 TI - Effects of interventions for the treatment of nocturnal enuresis in children. AB - The effectiveness of interventions for the treatment of nocturnal enuresis in children published in a recent issue of Effective Health Care is reviewed. PMID- 14532375 TI - Proceedings of the 4th International Meeting on Calcitonin Gene-Related Peptide and its Receptor. AB - Calcitonin Gene-Related Peptide (CGRP), a 37 amino acid peptide identified as the alternately spliced gene product of calcitonin gene, is a sensory neuropeptide with potent cardiovascular effects. CGRP is distributed throughout the central and peripheral nervous systems and possesses diverse biological actions. CGRP has been suggested to play a role in diseases such as migraine, diabetes, pain, and inflammation. Two forms of CGRP (alpha and beta) that differ in three amino acids have been identified and are encoded by different genes. Based on the differential biological activities of various CGRP analogs, the CGRP receptors have been classified into CGRP1 and CGRP2. Structure-activity studies of CGRP analogs showed that the C- and N-terminal regions of the peptide interact independently with their receptors. While C-terminal peptide, CGRP (8-37) behaves as a CGRP1 receptor antagonist, N-terminal peptide CGRP (1-12) behaves as a weak agonist. Structural modifications of CGRP(28-37) have yielded micromolar to nanomolar affinity ligands. CGRP receptor belongs to the calcitonin receptor like receptor (CRLR) family of G-protein-coupled receptors and has been shown to require a single transmembrane domain protein called receptor activity modifying protein-1 (RAMP1) for its functional expression as well as activity. Human, rat, and porcine CRLRs have been cloned and characterized. Currently, the major focus is on the identification of potent and specific nonpeptide antagonists for this receptor in order to understand the physiological and pathophysiological role of this peptide. PMID- 14532423 TI - Computer-assisted reconstruction and motion analysis of the three-dimensional cell. AB - Even though several microscopic techniques provide three-dimensional (3D) information on fixed and living cells, the perception persists that cells are two dimensional (2D). Cells are, in fact, 3D and their behavior, including the extension of pseudopods, includes an important 3D component. Although treating the cell as a 2D entity has proven effective in understanding how cells locomote, and in identifying defects in a variety of mutant and abnormal cells, there are cases in which 3D reconstruction and analysis are essential. Here, we describe advanced computer-assisted 3D reconstruction and motion analysis programs for both individual live, crawling cells and developing embryos. These systems (3D DIAS, 3D-DIASemb) can be used to reconstruct and motion analyze at short time intervals the nucleus and pseudopodia as well as the entire surface of a single migrating cell, or every cell and nucleus in a developing embryo. Because all images are converted to mathematical representations, a variety of motility and dynamic morphology parameters can be computed that have proven quite valuable in the identification of mutant behaviors. We also describe examples of mutant behaviors in Dictyostelium that were revealed through 3D analysis. PMID- 14532424 TI - Quality of life as medicine. II. A pilot study of a five-day "quality of life and health" cure for patients with alcoholism. AB - Alcoholism can be understood as a self-treatment for existential pain. A 5-day treatment was designed to relieve this psychological pain and existential anxiety, and thereby diminish the need for self-treatment with alcohol. The basic principle behind the treatment was holistic, restoring the quality of life (QOL) and relationship with self, which according to the life mission theory happens when life-denying views are corrected and inner emotional conflicts are solved. The method in this treatment was a course with teachings in philosophy of life, psychotherapy, and body therapy. The synergy attained was considerable and the outcome demonstrates that in the course of 1 week, people have time to revise essential life-denying views and to integrate important, unfinished life events involving negative feelings. This was demonstrated by an improved QOL and a decrease in their dependency and need for alcohol abuse. In the week before, after the 5-day course, and again after 1 and 3 months, the 16 participants completed the SEQOL questionnaire on QOL and health. This was a pilot study based on a pre-experimental design, without a control group and without clinical control. Common for the group were a low QOL, numerous health problems, and alcohol dependency in spite of treatment with Antabus (disulfiram). The study showed an increase in QOL from 57.6% before the course to 69.4% 3 months after the course, or an improvement in QOL of 11.8%. There was a 24.0% improvement in self-perceived mental health, and satisfaction with health in general was improved by 11.1%. The total sum of health symptoms in the group was reduced from 59% of maximum to 33%. It is concluded that for this small and motivated group with alcohol problems, it was possible to improve QOL and health in only 5 days with a holistic treatment that combined philosophy of life, psychotherapy, and body therapy, but the results are not final. Further research is needed. PMID- 14532425 TI - Modern approach to ureteral stones. AB - Urolithiasis is a very common affliction of mankind. In western countries incidence is increasing steadily. An increasing proportion of patients are presenting with ureteral stones, of which renal colic most often is the first complaint and the most common reason for an emergency visit to a urologist. Proper imaging strategy is of paramount importance in the diagnosis of acute flank pain and in the subsequent therapy planning once a ureteral stone is diagnosed. Renal colic during pregnancy poses specific problems, both in imaging and therapy. Apart from the adequate treatment of renal colic, modern therapy of those ureteral calculi that will not pass spontaneously will consist of a judicious combination of ESWL (extracorporeal shock wave lithotripsy), endourology, and laparoscopy. Open surgery should only be reserved for limited and very specific indications. Although beyond the scope of this article, metaphylaxis should take an important role in the follow-up of stone patients in general. PMID- 14532427 TI - Rabs, Rips, FIPs, and endocytic membrane traffic. AB - Rab GTPases, proteins belonging to the Ras-like small GTP-binding protein superfamily, have emerged as master regulators of cellular membrane transport. Rab11 GTPase, a member of the Rab protein family, plays a role in regulating various cellular functions, including plasma membrane recycling, phagocytosis, and cytokinesis. Rab11 acts by forming mutually exclusive complexes with Rab11 family binding proteins, known as FIPs. Rab11-FIP complexes serve a role of "targeting complexes" by recruiting various membrane traffic factors to cellular membranes. Recent studies have identified several Rab11-FIP complex-binding proteins that regulate distinct membrane traffic pathways. PMID- 14532428 TI - Voluntary spatial attention has different effects on voluntary and reflexive saccades. AB - Although numerous studies have investigated the relationship between saccadic eye movements and spatial attention, one fundamental issue remains controversial. Some studies have suggested that spatial attention facilitates saccades, whereas others have claimed that eye movements are actually inhibited when spatial attention is engaged. However, these discrepancies may be because previous research has neglected to separate and specify the effects of attention for two distinct types of saccades, namely reflexive (stimulus-directed) and voluntary (antisaccades). The present study explored the effects of voluntary spatial attention on both voluntary and reflexive saccades. Results indicate that voluntary spatial attention has different effects on the two types of saccades. Antisaccades were always greatly facilitated following the engagement of spatial attention by symbolic cues (arrows) informing the subject where the upcoming saccade should be directed. Reflexive saccades showed little or no cueing effects and exhibited significant facilitation only when these cues were randomly intermixed with uncued trials. In addition, the present study tested the effects of fixation condition (gap, step, and overlap) on attentional modulation. Cueing effects did not vary due to fixation condition. Thus, voluntary spatial attention consistently showed different effects on voluntary and reflexive saccades, and there was no evidence in these studies that voluntary cues inhibit reflexive saccades, even in a gap paradigm. PMID- 14532645 TI - Tetraaquacobalt(II) bis[vanadyl(IV) phosphate], [Co(H(2)O)(4)][VO(PO(4))](2). AB - The structure of [Co(H(2)O)(4)][VO(PO(4))](2) is composed of [VO(PO(4))] layers and interlayer tetrahydrated Co(2+) ions. Alternating VO(5) square pyramids and PO(4) tetrahedra share O-atom vertices, thus forming the vanadyl phosphate layers. Two vanadyl oxo groups from neighbouring layers are coordinated to each Co atom in a trans fashion, with Co-O distances of 2.157 (4) A, thus generating a three-dimensional framework structure. PMID- 14532646 TI - Cs[Si(3)O(6)(OH)] and Rb[Si(2)O(4)(OH)]: two novel phyllosilicates. AB - The crystal structures of two novel phyllosilicates with compositions Cs[Si(3)O(6)(OH)] (caesium hydroxohexaoxotetraotrisilicate) and Rb[Si(2)O(4)(OH)] (rubidium hydroxohexaoxotetraodisilicate) have been characterized by X-ray diffraction. The topology of the caesium phyllosilicate silica sheet consists of interconnected four- and six-membered rings and thus differs from all of the previously reported phyllosilicates. The topology of the rubidium phyllosilicate silica sheet consists of six-membered rings only, in boat conformations, resulting in a corrugated sheet similar to that observed in delta-Na(2)Si(2)O(5). Both of the title compounds exhibit the characteristic sandwich structure of sheet silicates, with the Cs atom ninefold coordinated and the Rb atom eightfold coordinated to the framework O atoms. PMID- 14532647 TI - beta-Y(2)Si(2)O(7), a new thortveitite-type compound, determined at 100 and 280 K. AB - A new form of Y(2)Si(2)O(7) (diyttrium heptaoxodisilicate) has been synthesized which is isotypic with thortveitite, Sc(2)Si(2)O(7), and crystallizes in the centrosymmetric space group C2/m, both at 100 and 280 K. The Y(3+) cation occupies a distorted octahedral site, with Y-O bond lengths in the range 2.239 (2)-2.309 (2) A. The SiO(4) tetrahedron is remarkably regular, with Si-O bond lengths in the range 1.619 (2)-1.630 (2) A. The bridging O atom of the Si(2)O(7) pyrosilicate group shows a large anisotropic displacement perpendicular to the Si O bond. Changes in lattice and structural parameters upon cooling are small with, however, a distinct decrease of the anisotropic displacement of the briding O atom. Structure solution and refinement in the non-centrosymmetric space group C2 are possible but do not yield a significantly different structure model. The Si-O Si bond angle of the isolated Si(2)O(7) groups is 179.2 (1) degrees at 280 K in C2 and 180 degrees per symmetry in C2/m. The C2/m structure model is favoured. PMID- 14532648 TI - Tris(3,5-dimethyl-1H-pyrazole-1-thiocarboxamidato-kappa(2)N(2),N)cobalt(III). AB - In the crystal structure of the title complex, [Co(C(6)H(8)N(3)S)(3)], the Co(III) atom is octahedrally coordinated by three monodeprotonated bidentate 3,5 dimethyl-1H-pyrazole-1-thiocarboxamide ligands with two thiocarboxamide N atoms in axial positions. The asymmetric unit contains two molecules (A and B) and these molecules are arranged in chains in an alternating fashion connected by N H...S interactions. PMID- 14532649 TI - A uranyl ion complex of N-methyl-p-tert-butyldihomoammoniocalix[4]arene with diaquadipyridinelithium as counter-ion. AB - The title complex, diaquadipyridinelithium (N-methyl-p-tert butyldihomoammoniocalix[4]arene-kappa(4)O)dioxouranium(VI) tripyridine solvate monohydrate, [Li(C(5)H(5)N)(2)(H(2)O)(2)][UO(2)(C(46)H(58)NO(4))].3C(5)H(5)N.H(2)O, contains an 'internal' tetraphenoxide-coordinated uranyl complex of the macrocycle, in which the protonated N atom is involved in an intramolecular hydrogen bond with the uranyl oxo group located in the cavity. The Li(+) ion is in a tetrahedral environment and its two water ligands are involved in hydrogen bonds with two phenoxide O atoms, two pyridine molecules and one water molecule. This arrangement is compared with those obtained previously for other homoazacalixarenes and also for homooxacalixarenes in the presence of alkali metal hydroxides. PMID- 14532650 TI - Helical catena-poly[[tris(1H-benzimidazole-kappaN(3))cobalt(II)]-micro-maleato kappa(3)O,O':O"]. AB - The crystal structure of the title compound, [Co(C(4)H(2)O(4))(C(7)H(6)N(2))(3)](n), consists of polymeric chains of the Co(II) complex. Two maleate dianions and three benzimidazole ligands coordinate to the Co(II) atom with a distorted octahedral geometry. The maleate dianions bridge neighbouring Co(II) atoms via both terminal carboxylic acid groups, one of which is monodentate and the other bidentate, to form a helical structure of alternating maleate dianions and Co(II) atoms, with a pitch height of 9.2667 (17) A. The absolute structure has been determined, and the crystal contains only right-handed helices. Intrahelical N-H...O hydrogen bonds stabilize the helical structure, while interhelical N-H...O hydrogen bonds link neighbouring helices to form the supramolecular structure. PMID- 14532651 TI - Tris[diphenyl(2-pyridyl)phosphine]-micro(3)-iodo-tri-micro(3)-sulfido sulfidotrisilvertungsten dichloromethane hemisolvate. AB - The title compound, [Ag(3)WIS(4)(C(17)H(14)NP)(3)].0.5CH(2)Cl(2), is a cubane type heterometallic cluster containing diphenyl(2-pyridyl)phosphine (Ph(2)PPy). The pyridyl group of Ph(2)PPy remains uncoordinated, so the Ph(2)PPy ligand is monodentate and coordinates to one Ag atom. The W atom and three Ag atoms form a distorted tetrahedral geometry, capped by one I atom and three S atoms. PMID- 14532652 TI - Bis(dicyanamido)(diethylenetriamine-kappa(3)N)copper(II) and (dicyanamido)(triethylenetetramine-kappa(4)N)copper(II) dicyanamide. AB - Two new complexes, [Cu(C(2)N(3))(2)(dien)] (dien is diethylenetriamine, C(4)H(13)N(3)), (I), and [Cu(C(2)N(3))(trien)](C(2)N(3)) (trien is triethylenetetramine, C(6)H(18)N(4)), (II), have been characterized by single crystal X-ray diffraction. Both complexes display a distorted tetragonal pyramidal geometry. In (I), the Cu atom is coordinated in the basal plane by three diethylenetriamine N atoms [Cu-N = 2.000 (2), 2.004 (2) and 2.025 (2) A] and one terminal N atom [Cu-N = 1.974 (2) A] from one monodentate dicyanamide group, and in the apical position by one terminal N atom [Cu-N = 2.280 (2) A] from the other monodentate dicyanamide group. In (II), the Cu atom is surrounded by four triethylenetetramine N atoms [Cu-N = 2.012 (2), 2.014 (2), 2.019 (2) and 2.031 (2) A in the basal plane] and a terminal N atom [Cu-N = 2.130 (2) A in the apical site] from one monodentate dicyanamide group. The other dicyanamide anion is not directly coordinated to the metal atom. In both (I) and (II), hydrogen bond interactions between the uncoordinated terminal N atoms of two dicyanamide ions and the amine H atoms lead to the formation of three-dimensional networks. PMID- 14532653 TI - Two isomorphous imidazole (Him) complexes: [MCl(2)(Him)(2)(H(2)O)(2)] (M = Co and Ni). AB - The structures of aquadichlorobis(1H-imidazole)cobalt(II), [CoCl(2)(Him)(2)(H(2)O)(2)] (Him is 1H-imidazole, C(3)H(4)N(2)), (I), and aquadichlorobis(1H-imidazole)nickel(II), [NiCl(2)(Him)(2)(H(2)O)(2)], (II), are isomorphous and consist of monomers with inversion symmetry. The three monodentate ligands (imidazole, chlorine and aqua), together with their symmetry equivalents, define almost perfect octahedra. Hydrogen-bonding interactions via the imidazole and aqua H atoms lead to a three-dimensional network. PMID- 14532654 TI - Tris(1,10-phenanthroline)sodium 2,4,6-trimercapto-1,3,5-triazin-1-ide. AB - The title compound, [Na(C(12)H(8)N(2))(3)](C(3)H(2)N(3)S(3)), contains an Na(+) centre which is ionicly bonded to three 1,10-phenanthroline (phen) ligands and one trithiocyanurate(1-) (ttcH(2)) anion. In the crystal structure, the anions are linked via hydrogen bonds to form linear chains. The S and H atoms of the ttcH(2) anion participate in intermolecular N-H.S hydrogen bonding, with N.S distances of 3.298 (2) and 3.336 (2) A. The phen ligands are almost parallel, with dihedral angles of 3.92 (5), 11.75 (5) and 15.45 (5) degrees; moreover, they are nearly perpendicular to the ttcH(2) chains, with angles of 81.94 (7), 85.86 (7) and 85.96 (7) degrees. PMID- 14532655 TI - catena-poly[[[(1,10-phenanthroline-kappa(2)N,N')manganese(II)]-microL-tartrato kappa(4)O(1),O(2):O(3),O(4)] hexahydrate]. AB - The title compound, [[Mn(C(4)H(4)O(6))(C(12)H(8)N(2))].6H(2)O](n), has a linear chain structure containing monomeric [Mn(C(4)H(4)O(6))(C(12)H(8)N(2))] repeat units. Each manganese ion is six-coordinate, with the two phenanthroline N atoms [Mn-N = 2.229 (2) and 2.235 (2) A] and four O atoms from two tartrate anions [Mn O(COO) = 2.1252 (19) and 2.1310 (19) A, and Mn-O(OH) = 2.2404 (19) and 2.2424 (19) A] forming a seriously distorted octahedral coordination environment. Six water molecules exist outside every repeat unit as solvate molecules. Extensive hydrogen-bonding interactions and pi-pi stacking of the phenanthroline moieties exist between the chains. PMID- 14532656 TI - Tetra-microchloro-1:2kappa(4)Cl;1:3kappa(4)Cl-bis(4,4'-dimethyl-2,2'-bipyridine) 2kappa(2)N,3kappa(2)N-lithium(I)dipalladium(II) tetrakis(pentafluorophenyl)borate dichloromethane 1.196-solvate. AB - In the crystal structure of the title compound, [LiPd(2)Cl(4)(C(12)H(12)N(2))(2)](C(24)F(20)B).1.196CD(2)Cl(2) or [[(Me(2)bipy)PdCl(2)](2)(micro-Li)](+).B(C(6)F(5))(4)(-).1.196CD(2)Cl(2) (Me(2)bipy is 4,4'-dimethyl-2,2'-bipyridine), an Li(+) cation is stabilized by complexation with two (Me(2)bipy)PdCl(2) units through weak Li-Cl interactions. This compound is thus a rare example of a complex that exhibits an arrested Cl(-) abstraction. PMID- 14532657 TI - The isothiocyanate complex of triphenylborane forms an unusual coordination polymer with [K(18-crown-6)](+), both in the solid state and in solution. AB - The title salt, (1,4,7,10,13,16-hexaoxacyclooctadecane kappa(6)O)[(isothiocyanato)triphenylborato-kappaS]potassium(I), [K(C(19)H(15)BNS)(C(12)H(24)O(6))] or [K(SCNBPh(3))(18-crown-6)], where 18-crown 6 is 1,4,7,10,13,16-hexaoxacyclooctadecane and [SCNBPh(3)](-) is the (isothiocyanato)triphenylborate anion, exhibits a supramolecular structure that is best described as a helical coordination polymer or molecular screw. This unusual supramolecular structure is based on a framework in which the SCN(-) ion bridges the chelated K(+) ion and the B atom of BPh(3) in a micro(2) fashion. The X-ray crystal structure of the title salt has been determined at 100 (1) and 293 (2) K. The K(+) ion exhibits axial ligation by the S atom of the [SCNBPh(3)](-) anion, with a K-S distance of 3.2617 (17) A (100 K). The trans-axial ligand is an unexpected eta(2)-bound C=C bond of a phenyl group (meta- and para-C atoms) that belongs to the BPh(3) moiety of a neighboring molecule. The K-C bond distances span the range 3.099 (3)-3.310 (3) A (100 K) and are apparently retained in CDCl(3) solution (as evidenced by (13)C NMR spectroscopy). By virtue of the latter interaction, the supramolecular structure is a helical coordination polymer, with the helix axis parallel to the b axis of the unit cell. IR spectroscopy and semi-empirical molecular orbital (AM1) calculations have been used to investigate further the electronic structure of the [SCNBPh(3)](-) ion. PMID- 14532658 TI - Bis(acetonitrile-kappaN)[(1RS,4RS,8SR,11SR)-1,4,8,11-tetraazatetradecane kappa(4)N]copper(II) bis[tetrakis[3,5-bis(trifluoromethyl)phenyl]borate] 0.31 hydrate. AB - The title compound, [Cu(C(2)H(3)N)(2)(C(10)H(24)N(4))](C(32)H(12)BF(24))(2).0.31H(2)O, crystallizes as an ionic species with no interactions between the ions. The [Cu(II)(cyclam)(MeCN)(2)](2+) dication (cyclam is 1,4,8,11-tetraazatetradecane), located on a 2/m symmetry site, forms as a distorted octahedral species with four Cu-N(cyclam) bonds of 2.013 (2) A and two C-N(MeCN) bonds of 2.499 (4) A. The [B[C(6)H(3)(CF(3))-3,5](4)](-) anion, located on a twofold axis, is a distorted octahedral species. A small amount of water is present, occupying sites between columns of ions. PMID- 14532659 TI - Three yttrium crotonate complexes with diimines. AB - The synthesis and crystal structures of three new yttrium crotonate (crot) compounds, associated with three different nitrogenous bases, namely 1,10 phenanthroline (phen), 4-methyl-1,10-phenanthroline (mphen) and 2,2' bipyridylamine (bpa), are presented. All three compounds organize as centrosymmetric dimers, to give tetra-micro-crotonato-bis[crotonato(1,10 phenanthroline)yttrium(III)] dihydrate, [Y(2)(C(4)H(5)O(2))(6)(C(12)H(8)N(2))(2)].2H(2)O or [Y(crot)(3)(phen)](2).2H(2)O, (I), tetra-micro-crotonato-bis[crotonato(4-methyl-1,10 phenanthroline)yttrium(III)] dihydrate, [Y(2)(C(4)H(5)O(2))(6)(C(13)H(10)N(2))(2)].2H(2)O or [Y(crot)(3)(mphen)](2).2H(2)O, (II), and tetra-micro-crotonato bis[diaqua(crotonato)yttrium(III)] 2,2'-bipyridylamine tetrasolvate, [Y(2)(C(4)H(5)O(2))(6)(H(2)O)(4)].4C(10)H(9)N(3) or [Y(crot)(3)(aq)(2)](2).4(bpa), (III). Complexes (I) and (II) are isomorphous, with the bases acting as chelating ligands. In complex (III), the coordination sphere is built up of carboxylate and aqua ligands, with the non-coordinated diimine acting as included solvent. PMID- 14532660 TI - Potassium 3-cyano-4-(dicyanomethylene)-5-oxo-4,5-dihydro-1H-pyrrol-2-olate. AB - The crystal structure of the title potassium salt, K(+).C(8)HN(4)O(2)(-), of the organic anion 3-cyano-4-(dicyanomethylene)-5-oxo-4,5-dihydro-1H-pyrrol-2-olate shows that the dicyanomethylene moiety is able to accept an electron in the same way as does tetracyanoethylene, to yield the novel product. The organic anion is nearly planar, with deviations caused by steric crowding among the exocyclic cyano groups. The K(+) cations lie within tricapped trigonal prisms that stack to form channels. The three-dimensional structure is completed by the formation of hydrogen-bonded chains by the anions. PMID- 14532661 TI - [1,3-Bis(4-nitrophenyl)triazenido](triphenylphosphine)gold(I). AB - In the title complex, [Au(C(12)H(8)N(5)O(4))(C(18)H(15)P)], the coordination geometry about the Au(I) ion is linear, with one deprotonated 1,3-bis(4 nitrophenyl)triazenide ion, [O(2)NC(6)H(4)N=N-NC(6)H(4)NO(2)](-), acting as a monodentate ligand (two-electron donor), and one neutral triphenylphosphine molecule completing the metal coordination. The triazenide ligand is almost planar (r.m.s. deviation = 0.0767 A), with the largest interplanar angle being 11.6 (7) degrees between the phenyl ring of one of the terminal 4-nitrophenyl substituents and the plane defined by the N=N-N triad. The Au-N and Au-P distances are 2.108 (5) and 2.2524 (13) A, respectively. Pairs of molecules generated by centrosymmetry are associated into a supramolecular array via intermolecular C-H...O interactions, and N...C and N...O pi-pi interactions. PMID- 14532662 TI - Isolated pentaiodide anions in [K(18-crown-6)]I(5). AB - (1,4,7,10,13,16-Hexaoxacyclooctadecane-kappa(6)O)potassium pentaiodide, [K(C(12)H(24)O(6))]I(5), obtained by slow evaporation of an ethanol solution of KI, 18-crown-6 and I(2), contains [K(18-crown-6)](+) cations (C(i) symmetry) and I(5)(-) anions (C(2) symmetry), which are arranged in alternating layers parallel to (001). In contrast to the well known tendency of I(5)(-) ions to form chains and nets, the I(5)(-) units in the title compound are isolated. PMID- 14532663 TI - ap-9-(meta-tert-butylphenyl)fluorene. AB - The title compound, C(23)H(22), (I), crystallizes in an ap conformationThe designations sp (synperiplanar) and ap (antiperiplanar) for these fluorene rotamers are in accordance with Rule E-6.6, IUPAC Tentative Rules, Section E, Fundamental Stereochemistry [J. Org. Chem. (1970), 35, 2861]. and its melt readily recrystallizes on cooling, in contrast to the corresponding 9-fluorenol compound, (II), which is sp and which melts without decomposition and fails to recrystallize over a long period. Both of these differences are ascribed to the intermolecular hydrogen bonding in (II), which is absent in (I) and which leads to distinctly different molecular packing in the two compounds. PMID- 14532664 TI - Ethyl N-phenyloxamate. AB - The oxamate group in the title compound, C(10)H(11)NO(3), is almost coplanar with the phenyl ring because of intramolecular hydrogen-bonding interactions, and the structure can be described as an anilide single bonded to an ethyl carboxylate group. The supramolecular structure is achieved through intermolecular hard N H...O and soft C-H...X (X = O and phenyl) hydrogen-bonding interactions. PMID- 14532665 TI - 2-Amino-1,3-benzothiazole-ethyl coumarin-3-carboxylate (1/1). AB - The title adduct, C(7)H(6)N(2)S.C(12)H(10)O(4), is formed via N-H.O and N-H.N hydrogen-bonding interactions, which generate a tetrameric unit with a pseudo centre of symmetry. The tetramer further packs through parallel-displaced pi-pi stacking interactions along the a direction. PMID- 14532666 TI - Benzyltripropylammonium bromide: a structure in polar space group P4(2)bc. AB - Crystals of the title compound, C(16)H(28)N(+).Br(-), were grown from solution in a mixture of acetone and propan-2-ol by slow evaporation. The structure was solved in the polar space group P4(2)bc with five moieties in the asymmetric unit, namely two benzyltripropylammonium cations in general positions, and two Br(-) anions in special Wyckoff positions (4a2. and 4b2.) and one in a general position. The structure consists of two kinds of molecular columns parallel to c, built of cations connected through C-H.pi hydrogen bonds and stabilized by weak C H.Br interactions. PMID- 14532667 TI - 1,2-dinitroguanidine. AB - The title compound, CH(3)N(5)O(4), is almost planar, and the conformation is fixed by two intramolecular N-H...O hydrogen bonds. Owing to the delocalization of pi-electron density over the whole molecule, there is through-conjugation, with the C-N, N-N and N-O bond lengths having values intermediate between those typical for the corresponding single and double bonds. PMID- 14532669 TI - C.I. Pigment red 266. AB - C.I. Pigment Red 266, or 4-[[4-(aminocarbonyl)phenyl]hydrazono]-N-(2 methoxyphenyl)-3-oxo-3,4-dihydronaphthalene-2-carboxamide, C(25)H(20)N(4)O(4), adopts the keto-hydrazone tautomeric form with significant intramolecular hydrogen bonding. The molecules pack to form layers involving an extensive network of intermolecular hydrogen bonds, in which the primary amide group plays a prominent role. The good technical performance of this pigment in application may be attributed principally to the pattern of intra- and intermolecular hydrogen bonding. PMID- 14532668 TI - Bis(glycylglycinium) oxalate at 100 K. AB - The structure of the title compound, 2C(4)H(9)N(2)O(3)(+).C(2)O(4)(2-), which has been determined by X-ray diffraction, contains discrete glycylglycine (HGly Gly)(+) cations in general positions and oxalate anions which lie across centres of inversion. Although the geometry of the (HGly-Gly)(+) cation is not significantly different compared with other structures containing this residue, a few changes in conformation are observed which indicate the presence of molecular interactions. The molecular network in the crystal consists of one nearly linear O-H...O, five N-H...O and two weak C-H.O hydrogen bonds. PMID- 14532670 TI - Crystal engineering with heteroboranes. II. 1,2-Dicarboxy-1,2-dicarba-closo dodecaborane(12) ethanol hemisolvate. AB - The title compound, 1,2-(COOH)(2)-1,2-closo-C(2)B(10)H(10).0.5C(2)H(6)O or C(4)H(12)B(10)O(4).0.5C(2)H(6)O, forms a tetramer by incorporating ethanol (solvent) molecules through hydrogen bonding. Two eight-membered rings [graph set R(2)(2)(8)] are formed by hydrogen bonding between two carboxylic acid groups, whereas two ten-membered rings [R(3)(3)(10)] are formed by hydrogen bonding between two carboxylic acid groups and the OH group of an ethanol molecule (solvent). Two crystallographically independent tetramers are present in the crystal structure. PMID- 14532671 TI - R4/4 (30) rectangular rings in 2,5-dioxopiperazine-1,4-diacetic acid. AB - Molecules of the title 2,5-dioxopiperazinedione derivative, C(8)H(10)N(2)O(6), occupy centres of symmetry in the crystal structure. The six-membered ring has an almost planar conformation, with the substituent on nitrogen nearly perpendicular to the ring. The ideal geometry of the isolated molecule, as determined by ab initio HF-LCAO quantum-mechanical calculations, is slightly more puckered than that observed in the solid state. In the crystal structure, a strong hydrogen bond joins neighbouring molecules, thus forming a network of rectangular R(4)(4)(30) rings. PMID- 14532672 TI - Rearrangement products of 3-methanesulfonyl-N-methyl-N-nitroaniline. AB - Two isomeric products (C(8)H(10)N(2)O(4)S) of the rearrangement of 3 methanesulfonyl-N-methyl-N-nitroaniline have been investigated, viz. 3 methanesulfonyl-N-methyl-2-nitroaniline, which was the main product of the rearrangement, and 5-methanesulfonyl-N-methyl-2-nitroaniline. In both molecules, the aromatic rings are appreciably deformed towards ortho-quinonoidal geometry by electronic and steric interactions. The crystal structure is stabilized, in both cases, by weak C-H.O hydrogen bonds. PMID- 14532673 TI - Three polymorphs (alpha, beta, and delta) of D-mannitol at 100 K. AB - In the monoclinic delta polymorph of D-mannitol, C(6)H(14)O(6), both the molecule and the packing have approximate twofold rotational symmetry. The P2(1) structure thus approximates space group C222(1), and the alpha' polymorph, previously reported in that space group, is almost certainly identical to the delta polymorph. However, torsion angles along the main backbone of the molecule deviate from twofold symmetry by as much as 7.4 (3) degrees and the hydrogen bonding pattern does not conform to the higher symmetry. The alpha polymorph reported here is identical to the previously reported kappa polymorph, and the low-temperature structure of the beta polymorph agrees well with previously reported room-temperature determinations. The range of C-O bond lengths over the three polymorphs is 1.428 (2)-1.437 (4) A, and the range of C-C distances is 1.515 (4)-1.5406 (19) A. The delta polymorph has the highest density of the three, both at room temperature and at 100 K. PMID- 14532674 TI - Tribromo(3,5-dimethyl-2-nitrophenyl-kappa(2)C(1),O)tellurium(IV), bromo(3,5 dimethyl-2-nitrophenyl-kappa(2)C(1),O)tellurium(II) and bromo(3,5-dimethyl-2 nitrosophenyl-kappa(2)C(1),O)tellurium(II). AB - All three title compounds, prepared from bis(3,5-dimethyl-2 nitrophenyl)ditellurium, exhibit high degrees of intramolecular Te-O coordination. Their Te-O distances increase in the order C(8)H(8)BrNOTe < C(8)H(8)BrNO(2)Te < C(8)H(8)Br(3)NO(2)Te, with distances of 2.165 (3), 2.306 (1) and 2.423 (6) A, respectively, indicating that C(8)H(8)BrNOTe may be more aptly described as 1-bromo-4,6-dimethyl-2,1,3-benzoxatellurazole. PMID- 14532675 TI - Pergolide mesylate form II. AB - A new polymorph of pergolide mesylate or 8beta-[(methylsulfanyl)methyl]-6 propylergoline methanesulfonate, C(19)H(27)N(2)S(+).CH(3)SO(3)(-), is reported. Pergolide mesylate form II crystallizes in the trigonal system, which is unique for ergot derivatives. Although the hydrogen-bond system in form II differs completely from that in form I, the conformation of the pergolide moiety in various related structures is very similar. PMID- 14532676 TI - O-(2-tert-butyl-6-dimethylthiocarbamoyl-4-methylphenyl) N,N-dimethylthiocarbamate dichloromethane solvate. AB - In the title compound, C(17)H(26)N(2)OS(2).CH(2)Cl(2), the C=S distances are 1.650 (4) and 1.679 (3) A, and the torsion angle between the planes of the thiocarbamate and carbonothioyl fragments is 54.4 (2) degrees. The steric and electronic effects that these substituents exert on one another determine the observed anti configuration with respect to the phenyl C atoms to which they are attached. PMID- 14532678 TI - Berberine formate-succinic acid (1/1). AB - The title compound [systematic name: 9,10-dimethoxy-2,3-methylenedioxy-5,6 dihydrodibenzo[a,g]quinolizinium formate-succinic acid (1/1)], C(20)H(18)NO(4)(+).CHO(2)(-).C(4)H(6)O(4), contains centrosymmetric pairs of almost planar berberine cations, and hydrogen-bonded (C(4)H(6)O(4).HCOO(-))(2) rings of succinic acid with formate anions, bonded by O-H...O hydrogen bonds with O...O distances of 2.4886 (15) and 2.5652 (16) A. Pairs of cations and molecules of succinic acid are connected by non-conventional weak C-H.O hydrogen bonds, with C...O distances of 3.082 (2) and 3.178 (2) A. PMID- 14532677 TI - (-)-2-(1,2,3,4,4a,5,6,7,8,8aalpha-Decahydro-4abeta,8alpha-dimethyl-7-oxo-2beta naphthyl)propionic acid: catemeric hydrogen bonding in a bicyclic sesquiterpenoid zeta-keto acid. AB - In the title compound, C(15)H(24)O(3), derived from a naturally occurring sesquiterpenoid, the asymmetric unit consists of two molecules differing by 167.4 (8) degrees in the rotational conformation of the carboxyl group. Each molecule aggregates separately with its own type as carboxyl-to-ketone hydrogen-bonding catemers [O...O = 2.715 (6) and 2.772 (6) A, and O-H...O = 169 and 168 degrees ]. This generates two crystallographically independent single-strand hydrogen bonding helices passing through the cell in the b direction, with opposite end-to end orientations. One intermolecular C-H...O=C close contact exists for the carboxyl group of one of the molecules. The structure is isostructural with that of a closely related unsaturated keto acid reported previously. PMID- 14532679 TI - Crystal engineering with heteroboranes. III. 2-Carboxy-1-methoxymethyl-1,2 dicarba-closo-dodecaborane(12). AB - The title compound, 1-CH(2)OCH(3)-2-COOH-1,2-closo-C(2)B(10)H(10) or C(5)H(16)B(10)O(3), forms a discrete centrosymmetric tetramer, via hydrogen bonding, involving two inner and two outer carborane molecules. One conventional eight-membered hydrogen-bonded ring [graph set R(2)(2)(8)] is formed between two carboxylic acid groups of the inner carboranes. This interaction is then supplemented by an open finite hydrogen bond (graph set D) between the ether O atom of the inner carborane and the carboxylic acid H atom of the outer carborane. PMID- 14532680 TI - L-methionyl-L-alanine: a dipeptide with hexagonal symmetry and Z' = 7. AB - The crystal structure of L-methionyl-L-alanine, C(8)H(16)N(2)O(3)S, is very similar to that of L-valyl-L-alanine [Gorbitz & Gundersen (1996). Acta Cryst. C52, 1764-1767] and other related dipeptides in space group P6(1), but there are seven molecules in the asymmetric unit. The Z value of 42 is the highest ever observed for a chiral molecule. PMID- 14532681 TI - Dibothrioclinin I and II, epimers from Gerbera piloselloides (L.) Cass. AB - Dibothrioclinin I and II, namely (+)-(11R,12S,25R,27S)- and (+/-) (11RS,12RS,25RS,27SR)-3,3,7,17,21-pentamethyl-4,12,18,26 tetraoxaheptacyclo[15.11.1.0(2,15).0(5,14).0(6,11).0(19,28).0(20,25)]nonacosa 5(14),6,8,10,19(28),20,22,24-octaene-13,27-dione, respectively, are C(30)H(28)O(6) epimers which are derived from two bothrioclinin moieties joined so as to create an additional six-membered ring. Structurally, the epimers differ only by inversion at one C atom of a central ring junction and the corresponding six-membered rings have similar conformations in each molecule, except for one ring adjacent to this inversion site. PMID- 14532682 TI - 2-bromo-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine, a new unexpected bifunctional building block for combinatorial chemistry. AB - The first reported structure of a pyridin-2-ylboron derivative, viz. the title compound, C(11)H(15)BBrNO(2), (I), is compared with its regioisomer 2-bromo-5 (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine, (II) [Sopkova-de Oliveira Santos, Lancelot, Bouillon & Rault (2003). Acta Cryst. C59, o111-o113]. Structural differences are observed, firstly in the orientation of the dioxaborolane ring with respect to the pyridine ring and secondly in the bond angles of the BO(2) group. These differences do not explain the experimentally observed differences in chemical reactivity between (I) and (II) but do confirm the relatively lower stability of (I). However, ab initio calculations of the HOMO (highest occupied molecular orbital) and LUMO (lowest unoccupied molecular orbital), based on the known crystal structures of the two compounds, show different distributions, which correspond to the differences observed during chemical reactions. PMID- 14532683 TI - Methyl (+/-)-1-ethyl-2-hydroxy-4-(4-methoxybenzoyl)-5-(4-methoxyphenyl)-3-oxo-2,3 dihydro-1H-pyrrole-2-acetate. AB - The title compound, C(24)H(25)NO(7), is a racemic mixture of 2,3-dihydro-1H pyrrol-3-ones. It crystallizes in the triclinic system, space group P1, with Z = 2. The asymmetric unit contains two enantiomorphic molecules and the structure is stabilized by hydrogen-bond contacts. PMID- 14532684 TI - 2-[(2-hydroxy-4-nitrophenyl)aminomethylene]cyclohexa-3,5-dien-1(2H)-one. AB - The title compound, C(13)H(10)N(2)O(4), adopts the keto-amine tautomeric form and displays an intramolecular N-H...O [N...O = 2.579 (2) A] and three intermolecular O-H...O [O...O = 2.561 (2) A] and C-H...O [C...O = 3.274 (2) and 3.318 (2) A] hydrogen bonds. The keto-amine structure is favoured by through-molecule conjugation between the hydroxy O atom and imine N atom. The dihedral angle between the planes of the two aromatic rings is 10.79 (4) degrees. PMID- 14532685 TI - 5-chloro-2-hydroxybenzophenone, featuring O-H...O, C-H...O, C-H...pi and pi-pi interactions. AB - Molecules of the title compound, C(13)H(9)ClO(2), contain an intramolecular O H...O hydrogen bond, and the two aromatic rings are inclined at 57.02 (3) degrees with respect to one another. The crystal structure is supported by C-H...O, C H...pi and pi-pi interactions. PMID- 14532686 TI - Receptor-mediated transport of lactoferrin into the cerebrospinal fluid via plasma in young calves. AB - Milk, especially colostrum, contains different kinds of macromolecules abundantly, such as immunoglobulin G (IgG), lactoferrin (Lf), transferrin (Tf), and growth factors. These are essential for the development and maintenance of health, which greatly depends on the absorption and transportation of macromolecules to the target organs. To evaluate the macromolecular transport, and concentrations in plasma and cerebrospinal fluid (CSF), colostrum was fed to newborn calves followed by milk and milk replacer, and maintained up to the 4th week under farm conditions. Plasma and CSF were collected at different times, and were analyzed for Lf, Tf, IgG and iron concentrations. Lf, Tf and IgG concentrations were steeply increased in plasma and CSF after colostrum feeding, and fluctuating patterns were observed during the experiments. Furthermore, intraduodenal administration of bovine Lf alone in young calf experiments revealed that the Lf concentration reached a peak at 4 hr, and was 7 and 4 times higher than preadministration in plasma and CSF, respectively. To explore the transport mechanism of Lf into CSF in young calves, epithelial membranes of the choroid plexus were prepared and a binding assay for Lf receptors (Lf-R) was carried out with 125I-Lf. The saturation kinetics revealed that the Bmax of epithelial membranes was 26.15 nmol/mg protein with a Kd of 0.11 microM, which also showed that Lf-R is saturable and specific. Scatchard plot transformation showed the presence of a single type of Lf-R in the choroid plexus. These results suggest that Lf is transported into the CSF through receptor mediated transcytosis in young calves, and that Lf may play an important role(s) in brain function. PMID- 14532687 TI - Pressure-flow relationship and longitudinal distribution of pulmonary vascular resistance in heartworm-infected dogs. AB - The pressure-flow relationships and the longitudinal distributions of pulmonary vascular resistance in normal and heartworm-infected (HWI) dogs were compared in an isolated, blood perfused preparation. The pulmonary circulation was partitioned into pulmonary arterial, middle, and venous segment based on the concept of a five element lumped model. The pulmonary arterial pressure-flow relationships were found to be non-linear and convex to the pressure axis in both normal and HWI lungs. The pressure-flow relationships of the pulmonary arterial and venous segment were linear and these slopes in the HWI lungs were significantly higher than the normal lungs. The pressure gradient of the middle segment was increased as flow increased at lower flow range, however, it was not increased during higher perfusion range in both lungs. At higher flow, the pressure gradient of the middle segment in the HWI lungs was significantly higher than the normal lungs. These results suggest that the ohmic resistance was almost equal to the sum of the two slopes of the pressure-flow relationships of the pulmonary arterial and venous segment because the pressure gradient of the middle segment was not altered as flow increased during higher perfusion rate. Because the slopes of the pressure-flow relationships of the pulmonary arterial and venous segment were increased with heartworm infection, the ohmic resistance of HWI lungs would be higher than normal lungs. The intercept pressure on the pressure axis of the linear portion of the pulmonary arterial pressure-flow relationship, a critical closing pressure, was regarded as pressure gradient of the middle segment during higher perfusing rate because the intercept pressures of pressure-flow relationships of pulmonary arterial and venous segment were almost equal to zero. Therefore, the critical closing pressure of HWI lungs would be higher than normal lungs. The pulmonary hypertension of filariasis appears to be due to an increase in ohmic resistance and elevated critical closing pressure. PMID- 14532688 TI - Migration of Strongyloides venezuelensis in rats after oral inoculation of free living infective larvae. AB - Strongyloides venezuelensis (SVZ) infection was chronologically monitored in 85 Sprague-Dawley rats (SDR), which were orally inoculated with approximately 1,000 infective larvae. In order to describe the characteristics of migrating larvae (MLS) in various visceral organs (the liver, lung, cardiac blood, and small intestine), 5 SDR were sacrificed at 20 min, 45 min, 1 hr, 2 hr, 3 hr, 4 hr, 8 hr, 12 hr, 16 hr, 48 hr, 72 hr, 96 hr, 120 hr, 144 hr, 168 hr and 192 hr post inoculation (PI). MLS were recovered from the liver and blood 20 and 45 min PI and measured 788 +/- 26 microm and 846 +/- 40 microm in length, respectively. MLS were first observed in the lung tissue 45 min PI and measured 925 +/- 38 microm on the average. In the trachea, MLS measuring 849 +/- 75 microm appeared 3 to 96 hrs PI. Adult worms (AWS) measuring 1,926 +/- 521 microm to 2,956 +/- 159 microm in length were observed in the small intestine from 120 hr PI. The worms appeared to mature more than 168 hr PI and attained the average maximum length of 2,420 +/ 532 microm. At 3 hr PI focal hyperemic and necrotic lesions were evidently observed in the liver and lung, together with eosinophilic infiltration in the stomach, liver, and lung. The parasites were histologically detectable in the lung tissues but were very difficult to find in the liver and the epithelial layer of small intestine. These data demonstrate that SVZ parasites take 20 min to reach the liver via the stomach and only three hours to reach the trachea through the same route. The development from eggs to adults takes 168 hr in the SDR model. PMID- 14532689 TI - Cerebrospinal setariosis with Setaria marshalli and Setaria digitata infection in cattle. AB - Setaria digitata and S. marshalli larvae were observed in the cerebrospinal cavity of 2 paralyzed cattle in Taiwan. The 2 affected cattle showed quadriplegia and lumbar paralysis, respectively. At necropsy, which was performed 7 days after the 7-month-old cattle became quadriplegic, three and nineteen S. marshalli larvae as well as two female adult worms were found in the cranial cavity, spinal cavity and peritoneal cavity of the cattle, respectively. Necropsy on the other 8 month-old cattle was also performed 3 days after it showed lumbar paralysis, and ten S. digitata larvae were found in the spinal cavity. In both cattle, many mononuclear inflammatory cells mixed with a few eosinophils were seen accumulated in the connective tissue around the root of the spinal nerves. Infiltration of eosinophils and mononuclear inflammatory cells into the epidura and arachnoidea of the brain were also observed. The major inflammatory cell was lymphocytes, but neutrophils and eosinophils were also present. The number of cells in the cerebrospinal fluid collected initially from the two affected cattle were 105/0.01 ml and 143/ 0.01 ml, respectively. This is the first report of cerebrospinal setariosis in cattle associated with S. marshalli. PMID- 14532690 TI - Effects of a macrolide antibiotic on enamel formation in rat incisors--primary lesion of ameloblast at the transition stage. AB - A novel macrolide antibiotic was administered orally to 5-week-old Jcl:Wistar rats at a dose of 5,000 mg/kg/day for 5 weeks, and then a half of animals were maintained without any treatment for 10 weeks. A white discolored lesion with horizontal stripes developed on the surface of the upper and lower incisors after dosing for 4 weeks, and these macroscopical incisal lesions disappeared with the eruption in 4 weeks after stop of administration. Histopathologically, increase in number of karyopycnosis of ameloblast at the transitional stage, vacuolar degeneration of ameloblast and cystic change in the maturation stage, and impaired iron pigment secretion at the pigmentation stage were observed. Microradiography, calcio-traumatic zones, which means hypocalcification, were observed on the superficial layer of enamel. These results suggest that the primary lesion induced by a novel macrolide antibiotic is the increased karyopycnosis of ameloblast at the transitional stage, and followed by later stage. PMID- 14532691 TI - Effect of fusion/activation protocol on in vitro development of porcine nuclear transfer embryos constructed with foreign gene-transfected fetal fibroblasts. AB - The effect of fusion/activation protocol on in vitro development of porcine nuclear transfer (NT) embryos constructed with foreign gene-transfected somatic cells were investigated. NT embryos were produced by using enucleated M II oocytes and enhanced green fluorescence protein (EGFP) gene-transfected or non transfected porcine fetal fibroblasts. One group of NT embryos received a single electrical pulse to induce fusion and activation simultaneously (FAS). The other group was fused 2 hr before activation (FBA) using two kinds of electrical pulses. Electrically activated NT embryos in both groups were treated with cycloheximide (CHX) before culture to assess the development to the blastocyst stage. After 6 days of culture, all morulae and blastocysts derived from EGFP transfected fibroblasts emitted green fluorescence without mosaicism, and EGFP gene product was also detected in all morulae and blastocysts examined. NT embryos undergoing FAS showed higher developmental capacity to blastocysts than those undergoing FBA, regardless of the EGFP transfection into the nuclear donor cells. The results also indicated that EGFP-gene transfection into nuclear donor cells has no obvious deleterious effect on the development of NT embryos to blastocysts. PMID- 14532692 TI - Flow cytometric analysis of chicken NK activity and its use on the effect of restraint stress. AB - Immune system is organized by the influence of both neural and endocrine systems. NK activity plays an important role in the innate immunity. In this study, we observed the effects of restraint stress on chicken peripheral blood NK activity. Viability of FITC-labeled RP9 was measured with PI after treatment with the effector cells. Chicken peripheral blood CD8alpha+ cells expressed strong cytotoxic activity, in contrast to thrombocytes, while peripheral blood CD3+ CD8alpha+ cells and CD4+ cells had little cytotoxic activity. Con A supernatant enhanced the cytotoxic activity of CD8alpha+ cells. Therefore, it is considered that these cytotoxic activities measured by flow cytometry (FCM) analysis are NK activity. When chickens were exposed to restraint stress, the levels of serum corticosterone increased transiently over a short period of time while the NK activity decreased. The decreased NK activity, however, did not recover to the intact levels for a long time, even once the serum corticosterone levels had recovered. These data indicate that chicken NK activity is able to be measured by flow cytometric analysis and that restraint stress causes severe damage to the chicken NK activity. PMID- 14532693 TI - Seasonal changes in serum vitamin A, vitamin E and beta-carotene concentrations in Japanese Black breeding cattle in Hyogo prefecture. AB - The influence of the season on serum vitamin A, E and beta-carotene concentrations was studied in Japanese Black breeding cattle. Blood samples were collected from 13 cows once a month for a year to determine the serum vitamin concentrations. Mean serum concentrations (+/- SE) of retinol, alpha-tocopherol and beta-carotene were 21.3 +/- 0.5 microg/dl (range 9.9 to 44.9), 215.9 +/- 8.0 microg/dl (range 59.7 to 551.6) and 67.3 +/- 5.1 microg/dl (range 10.6 to 425.2), respectively. All these concentrations were lower than the recommended levels. Serum retinol and alpha-tocopherol concentrations in summer were lower than those in winter. These results suggest that the vitamin A, E and beta-carotene status of Japanese Black breeding cows is inadequate, and feeding high quality forages or dietary vitamin supplementation during summer should be recommended. PMID- 14532694 TI - Optimal timing for canine artificial insemination with frozen semen and parentage testing by microsatellite markers in superfecundency. AB - Parentage testing was performed in sixteen litters by canine artificial inseminations with frozen semen from different sires on Days 5 and 7 after the LH surge. It became apparent that only 25% of dams had superfecundation, but 43.8% of dams were whelped after insemination only on Day 5 after the LH surge and 31.3% of dams after insemination only on Day 7. Of the total 87 puppies, 46% were born after insemination on Day 5 after the LH surge and 54% after insemination on Day 7. This result strongly suggested that canine artificial insemination with frozen semen could be sufficiently successful also on Days 5 and 7 after the LH surge. PMID- 14532695 TI - Efficacy of enamel matrix protein applied to spontaneous periodontal disease in two dogs. AB - Enamel matrix protein (EMP) was applied for regeneration of periodontal tissue in 2 dogs with spontaneous periodontal disease. Case 1 had bony resorption around the root and root apex of the maxillary fourth premolars. Case 2 had vertical resorption of bone between the mandibular first and second molars. A flap was formed in the buccal gingiva, and EMP was applied onto the surface of the exposed root. One or 4 months postoperatively, increased bone level and clinical attachment were recognized. EMP was therefore suggested to be effective to induce regeneration of periodontal tissues in the cases with periodontal disease. PMID- 14532696 TI - Bronchoalveolar lavage fluid findings in the atelectatic regions of anesthetized horses. AB - To evaluate the effects of compression atelectasis on the composition of pulmonary secretions in anesthetized horses, cytological and biochemical examinations were performed on bronchoalveolar lavage (BAL) fluids obtained from both dependent and independent lung regions. Six horses were anesthetized with isoflurane and oxygen for 120 min, and were positioned in right lateral recumbency. Percentage of neutrophils and total protein concentration in BAL fluids significantly increased at the end of anesthesia, and total phosphorous concentration significantly decreased at 72 hr after anesthesia in dependent lung. Such changes were not observed within 168 hr after anesthesia. These findings suggest that the effects of compression atelectasis on the composition of pulmonary secretions may be eliminated within 168 hr after anesthesia. PMID- 14532697 TI - Seroepidemiologic studies on Babesia equi and Babesia caballi infections in horses in Jilin province of China. AB - The prevalence of equine piroplasmosis caused by Babesia equi and Babesia caballi in northeast China has remained unknown, although the People's Republic of China is recognized as an endemic country for the diseases. In the present study, we investigated the prevalence of equine piroplasmosis in Jilin province, a part of northeast China. A total of 111 serum samples were taken from horses in eastern Jilin, and examined for diagnosis of B. equi and B. caballi infections by the enzyme-linked immunosorbent assays with recombinant antigens, equi merozoite antigen-1 and P48, respectively. Of the 111 samples, 38 (34%) and 36 (32%) samples were sero-positive for B. equi infection and B. caballi infection, respectively. In addition, 14 (12%) samples were sero-positive for both B. equi and B. caballi infections. These results indicate that equine piroplasmosis is widespread and therefore a cause for serious concern in northeast China. PMID- 14532698 TI - Substances derived from 4-ethyl octanoic acid account for primer pheromone activity for the "male effect" in goats. AB - A major constituent of the characteristic "goaty odor" 4-ethyl octanoic acid (4EOA) was previously shown to have no primer pheromone activity. This was also confirmed by our own bioassay system utilizing the recording technique of neural activity of the hypothalamic gonadotropin-releasing hormone pulse generator in goats. However, when the synthetic 4EOA solution was kept at room temperature for several months, primer pheromone activity appeared in the same solution. Headspace gas chromatography/mass spectrometry analysis revealed that there were several newly formed substances in addition to 4EOA samples with primer pheromone activity. These results suggest that 4EOA derived substance(s) but not 4EOA itself is(are) primer pheromone in goats. PMID- 14532699 TI - Effects of alpha-pinene odor in different concentrations on stress-induced hyperthermia in rats. AB - Stress-induced hyperthermia is observed in animals exposed to stressful conditions. In our previous study, plant-derived fragrances such as green odor and alpha-pinene were shown to suppress this stress response in rats. In the present study, we examined the concentration-dependence of the alpha-pinene effects on stress-induced hyperthermia. Male rats carrying telemetry transmitters were transferred individually to a new cage containing bedding that had been sprayed with 0.3, 0.03, and 0.003% concentrations of alpha-pinene or control solvent. Following transfer to the novel environment, the body temperature increased significantly, and this response was clearly suppressed when the cage was scattered with 0.03% alpha-pinene only. These results suggest that the effect of alpha-pinene on stress-induced hyperthermia can be observed only at a certain concentration. PMID- 14532700 TI - The pathogenicity of canine parvovirus type-2b, FP84 strain isolated from a domestic cat, in domestic cats. AB - Canine parvovirus type-2a (CPV-2a) and type-2b (CPV-2b) have recently been isolated from domestic cats. The pathogenicity of CPV-2b in domestic cats is still unclear. In this study, we performed infection tests to examine the pathogenicity of CPV-2b, FP84 strain, isolated from a domestic cat. The results demonstrated that the CPV strain FP84 is able to infect and replicate well in domestic cats. Two of the 3 cats used in the test died. They showed loss of appetite, diarrhea, leukopenia and dehydration. Since FP84 was found to be virulent to domestic cats, it is necessary to examine the efficacy of inactivated feline panleukopenia virus vaccines against CPV infection in domestic cats. PMID- 14532701 TI - Optimization of in situ hybridization protocols for detection of feline herpesvirus 1. AB - In situ hybridization (ISH) protocol including microwaving pre-treatment regimes was developed and compared with protease digestion as a pre-treatment regime for its effects on detecting feline herpesvirus 1 (FHV-1) in formalin fixed, paraffin embedded tissue. We found that optimum results were obtained using microwave pre treatment. The results showed that the use of microwave irradiation would be recommended as a means of supplementing ISH methods, especially when using long term formalin fixed, paraffin-embedded tissue. PMID- 14532702 TI - Effects of intravenous infusion of hypotonic lactated Ringer's solution on calves with diarrhea. AB - The effects of intravenous infusion of hypotonic lactated Ringer's (LR: n=14) on plasma volume and venous blood gases were compared to those of hypotonic Ringer's solutions (RS: n=7) in diarrheic Japanese Black breed calves with metabolic acidosis. Venous blood samples were collected immediately before and after, and at 24 hr after the fluid infusion therapy. The LR and RS infusions increased relative plasma volume to 147.1 +/- 25.5% and 134.2 +/- 18.6%, respectively, just after the fluid therapy. The LR infusion induced an increase in the BE value (+5.1 +/- 4.8 mM) at 24 hr compared to that of RS. LR infusion should be explored as a treatment for dehydration and moderate metabolic acidemia caused by naturally occurring diarrhea in calves. PMID- 14532703 TI - Occurrence of ateliosis in Japanese Black calves and their secretory function of growth hormone. AB - Unthrifty calves occurred sporadically in Japanese Black (beef cattle) in an area in northeastern Japan. The states of unthrifty development, pedigree, clinico biochemistry and the secretory function of bovine growth hormone (bGH) in pituitary were investigated. The total cholesterol concentration and CK, AST and LDH activities in the serum showed higher values than those of control calves. Basal bGH concentrations in the serum and bGH secretory reactivity in the insulin tolerance test (ITT) were showed to be significantly lower than those of the control calves. Furthermore, sperm donated from a specific bull had been used for these unthrifty calves. This study suggested that the present occurrence of unthrifty calves represented ateliosis possibly caused by congenital hypopituitarism which decreased of bGH secretory function. PMID- 14532704 TI - Acute hepatitis in a piglet experimentally inoculated with tissue homogenates from pigs with postweaning multisystemic wasting syndrome. AB - Five 2 day-old colostrum-deprived piglets were inoculated with tissue homogenates from pigs with postweaning multisystemic wasting syndrome. One of the five piglets developed icterus and died 23 days post-inoculation. Histologic examination revealed acute hepatitis. Porcine circovirus type 2 (PCV-2) antigen and nucleic acid were detected in hepatocytes and phagocytic cells. Ultrastructurally, hepatocytes and phagocytic cells had large numbers of cytoplasmic inclusions, which were composed of electron-dense paracrystalline arrays of small non-enveloped viral particles approximately 17 nm in diameter. Apoptotic hepatocytes were confirmed by the TUNEL method and electron microscopic examination. These findings may indicate that hepatocellular necrosis is associated with replication of PCV-2. Apoptosis of hepatocytes also contributes to the pathogenesis of hepatic lesions in this case. PMID- 14532705 TI - Seroprevalence of Coxiella burnetii infections among cats in different living environments. AB - The seroprevalence of Coxiella burnetii infection among pet cats in Japan and Korea and stray cats in Japan was investigated by an indirect fluorescent antibody technique and PCR test. Forty-four (14.2%) of 310 pet cats in Japan were seropositive, as were 15 (41.7%) of 36 stray cats in Japan and 10 (8.6%) of 116 pet cats in Korea. The antibody positive rate in stray cats was significantly higher than that in pet cats, but there was no correlation between the rates in Japanese and Korean pet cats. In this study, the prevalence of C. burnetii infection among cats in different living environments was found and it is difficult to deny that stray cats would be one of the important sources of infection for human Q fever. PMID- 14532706 TI - Time scale of eutherian evolution estimated without assuming a constant rate of molecular evolution. AB - Controversies over the molecular clock hypothesis were reviewed. Since it is evident that the molecular clock does not hold in an exact sense, accounting for evolution of the rate of molecular evolution is a prerequisite when estimating divergence times with molecular sequences. Recently proposed statistical methods that account for this rate variation are overviewed and one of these procedures is applied to the mitochondrial protein sequences and to the nuclear gene sequences from many mammalian species in order to estimate the time scale of eutherian evolution. This Bayesian method not only takes account of the variation of molecular evolutionary rate among lineages and among genes, but it also incorporates fossil evidence via constraints on node times. With denser taxonomic sampling and a more realistic model of molecular evolution, this Bayesian approach is expected to increase the accuracy of divergence time estimates. PMID- 14532707 TI - Polymorphism at the esterase isozyme locus Est10 associated with phylogenetic differentiation in rice. AB - A new esterase isozyme locus, Est10, with 6 alleles including the null form, has been found in rice by using polyacrylamide gel electrophoresis. Thirty F(2) populations of all possible combinations between 5 different band morphs were studied. The segregation pattern indicated that bands 1, 2, 3, 4, and the null form (0) were allelic with each other. The alleles of Est10 were distributed at different frequencies among different varietal groups of rice and also between cultivated rice and its wild relatives (Oryza rufipogon Griff.). Alleles 1 and 2 were frequently found in Japonica and Indica types, respectively. Allele 3 showed a high frequency in Aus and Boro, both Indica types cultivated in South Asia. Allele 4 was frequent in wild rice O. rufipogon. Judging from the linkage between Est10 and RFLP marker RG220 and isozyme marker Est5, Est10 is located on chromosome 1. The importance of this locus in evolutionary studies of rice is discussed. PMID- 14532708 TI - Origin, dispersal and genomic structure of a low-copy-number hypervariable RFLP clone in Triticum and Aegilops species. AB - The genome of common wheat has evolved through allopolyploidization of three ancestral diploid genomes. A previously identified restriction fragment length polymorphism (RFLP) marker, pTag546, has the unique feature of showing hypervariability among closely related common wheat cultivars. To understand the origin and the mode of dispersal of this hypervariable sequence in the wheat genome, the distribution and structure of the homologous sequences were studied using ancestral diploid species, tetraploid disomic substitution lines and synthetic hexaploid lines. Comparative Southern blot and PCR analyses suggested that pTag546 homologs in the tetraploid and hexaploid wheat were derived from the S genome of Aegilops speltoides. Some pTag546 homologs were found to have transposed to A and D genomes in polyploid wheat. Evidence of transposition and elimination in some synthetic hexaploid lines was also obtained by comparing their copy numbers with those in the parental lines. Southern blot analysis of a genomic clone using a contiguous subset of sequences as probes revealed a core region of hypervariability that coincided with the region containing pTag546. No obvious structural characteristics that could explain the hypervariability, however, were found around the pTag546 sequence, except for accumulation of small repetitive sequences at one border. It was concluded that pTag546 increased its copy number through yet unknown mechanism(s) of transposition to various chromosomal locations over the period of allopolyploid evolution and during the artificial genome manipulation in wheat. PMID- 14532709 TI - Genomic distribution of three repetitive DNAs in cultivated hexaploid Diospyros spp. (D. kaki and D. virginiana) and their wild relatives. AB - To understand the genomic organization of Diospyros species with different ploidy levels, we cloned three different repetitive DNAs and compared their genomic distributions in ten Diospyros species, including hexaploid D. kaki and D. virginiana. Genomic Southern hybridization demonstrated that the EcoRV-repetitive DNA was present in tandem in the genomes of D. glandulosa (2n=2x=30), D. oleifera (2n=2x=30), D. lotus (2n=2x=30), D. virginiana (2n=6x=90) and D. kaki (2n=6x=90). All of these species except D. virginiana also contained the HincII-repetitive DNA in tandem. Fluorescent in situ hybridization showed that the EcoRV- and HincII-repetitive DNAs were predominantly located at the proximal or centromeric regions of chromosomes. The DraI-repetitive sequence cloned from D. ehretioides (2n=2x=30) was not found in the other Diospyros species tested. This suggests that D. ehretioides has a genomic organization different from that of the other Diospyros species. Speciation of hexaploid Diospyros species is also discussed with respect to the genomic distribution of the three repetitive DNAs cloned. PMID- 14532710 TI - Evolution of the GC content of the histone 3 gene in seven Drosophila species. AB - The molecular evolution of the histone multigene family was studied by cloning and determining the nucleotide sequences of the histone 3 genes in seven Drosophila species, D. takahashii, D. lutescens, D. ficusphila, D. persimilis, D.pseudoobscura, D. americana and D. immigrans. CT repeats, a TATA box and an AGTG motif in the 5' region, and a hairpin loop and purine-rich motifs (CAA(T/G)GAGA) in the 3' region were conserved even in distantly related species. In D. hydei and D.americana, the GC content at the third codon position in the protein coding region was relatively low (49% and 45%), while in D. takahashii and D. lutescens it was relatively high (64% and 65%). The non- significant correlation between the GC contents in the 3' region and at the third codon position as well as the evidence of less constraint in the 3' region suggested that mutational bias may not be the major mechanism responsible for the biased nucleotide change at the third codon position or for codon usage bias. PMID- 14532711 TI - [Current status of intrahepatic stones in Korea]. AB - Hepatolithiasis, or primary intrahepatic stones, is prevalent in the Far East. The majority of primary intrahepatic stones were calcium bilirubinate stones 30 40 years ago in Korea. However, intrahepatic stones with high cholesterol purity have recently been recognized. Since 1990s, mixed stones constituting about 40 50% of all intrahepatic stones, and pure intrahepatic cholesterol stones have been observed in Korea. Although advances in interventional radiological techniques, cholangioscopic technique, and novel surgical technique have led to significant changes in treating the patients with hepatolithiasis, it is still considered as an intractable disease and more common in Korea than western countries. Therefore, Korean gastroenterologists should have more interest and accomplish passionate researches on hepatolithiasis. PMID- 14532712 TI - [Prevalence of erosive esophagitis in patients with subtotal gastrectomy: case control study]. AB - BACKGROUND/AIMS: Enterogastric bile reflux has been implicated as a definite causative factor for the development of postoperative bile reflux gastritis. However, little is known about the role of bile reflux into the stomach in gastro esophageal reflux disease in patients with subtotal gastrectomy. In this study, we tried to prove that the bile reflux gastritis does increase the development of erosive esophagitis in patients who underwent subtotal gastrectomy. METHODS: From January 1997 to December 2001, 222 patients with previous subtotal gastrectomy were enrolled. We also reviewed the endoscopic findings in 1,633 age and sex matched healthy controls who had visited our hospital for routine check-up without significant gastrointestinal symptoms. RESULTS: The prevalence of erosive esophagitis is 5% (LA A 2.7%, LA B 2.3%) in gastrectomized patients and 4.9% (LA A 3.2%, LA B 1.7%, LA C 0.0%) in healthy controls. There was no significant difference in the prevalence and the degree of esophagitis between the two groups. CONCLUSIONS: These results suggest that subtotal gastrectomy may not be a risk factor for developing the erosive reflux esophagitis. PMID- 14532713 TI - [A prospective study on rabeprazole-based triple therapy for Helicobacter pylori eradication in patients with peptic ulcer]. AB - BACKGROUND/AIMS: Rabeprazole sodium is a potent proton pump inhibitor. We assessed the efficacy, safety and compliance of one-week triple therapy including rabeprazole with amoxicillin and clarithromycin for eradication of Helicobacter pylori (H. pylori). METHODS: Eighty-eight H. pylori-positive patients with peptic ulcer disease were received rabeprazole 10 mg bid, amoxicillin 1,000 mg bid and clarithromycin 500 mg bid for a week. Endoscopic examination with five biopsies (two specimens from the antrum, two from the gastric body, and one from the gastric angle) was performed. The status of H. pylori infection was assessed by histology (immunohistochemistry) of the biopsy specimens, 13C urea breath test, and CLO test at the beginning and 13C urea breath test 4 weeks after the completion of treatment. RESULTS: H. pylori eradication rates were 74.71% by intention-to-treat analysis and 87.84% by per-protocol analysis. The percentage of side effects was 12.5% and these side effects were not serious. CONCLUSIONS: One-week rabeprzole based triple therapy is an effective and safe regimen for H. pylori eradication in patients with peptic ulcer. PMID- 14532714 TI - [Helicobacter pylori infection in the remnant stomach after radical subtotal gastrectomy]. AB - BACKGROUND/AIMS: It was reported that the prevalence of Helicobacter pylori (H. pylori) infection decreased after gastrectomy, but persistent H. pylori infection may cause residual gastritis or stump cancer. We studied the prevalence of H. pylori infection in patients who had undergone subtotal gastrectomy for the treatment of gastric cancer and the factors that influence H. pylori positivity in the remnant stomach. METHODS: Ninety-eight patients who had undergone radical subtotal gastrectomy (RSG group) for the treatment of gastric cancer and eighty four patients diagnosed as having gastric cancer (GC group) were enrolled. H. pylori status was diagnosed by rapid urease test, histological examination, and 13C-urea breath test. We evaluated whether there were differences in various clinical characteristics according to the H. pylori status in the remnant stomach. RESULTS: The prevalences of H. pylori infection in RSG group and GC group were 55% and 69%, respectively. In RSG group, the prevalence of H. pylori was 76.9% in patients aged 49 or less, and it decreased with age. The prevalence of H. pylori within 3 years of gastrectomy was 59.5% and it decreased to 28.6% after 3 years of gastrectomy. CONCLUSIONS: The positive rate of H. pylori in RSG group is lower than that in GC group and decreases with age and time interval after operation. PMID- 14532715 TI - [Diagnosis of Helicobacter pylori infection with urine and stool]. AB - BACKGROUND/AIMS: The urine antibody and stool antigen test are newly developed non-invasive tests for Helicobacter pylori (H. pylori) infection. The aim of this study was to assess the diagnostic value of these tests. METHODS: Urine and stool specimens from 50 consecutive patients who had undergone gastroscopy (28 men, mean age 54 years) were obtained. A kit for antibody against H. pylori (RAPIRUN test) was used for urine specimens and a kit for H. pylori antigen using ELISA (HpSA test) was used for stool specimens. None of patients had ever received any treatment to eradicate H. pylori. The H. pylori status was evaluated based on three different tests (histology, 13C-urea breath test, rapid urease test) and defined as positive when two of three tests were positive. RESULTS: Of 50 patients, 31 patients were diagnosed as H. pylori positive and 15 patients were negative. The RAPIRUN test was positive in 24 of 31 patients (sensitivity 77.4%) and negative in 13 of 15 patients (specificity 86.7%). The HpSA test was positive in 25 of 31 patients (sensitivity 80.1%) and negative in 14 of 15 patients (specificity 93.3%). CONCLUSIONS: The accuracies of H. pylori urine antibody and stool antigen test are similar to those of Western studies. These two tests are found to be useful methods for diagnosis of H. pylori infection in Korea. PMID- 14532716 TI - [Characterization of pacemaking currents in cultured interstitial cells of Cajal from mice small intestine]. AB - BACKGROUND/AIMS: Gastrointestinal motility is initiated by the periodic generation of slow waves. Interstitial cells of Cajal (ICC) are pacemaker cells that generate slow waves and drive spontaneous mechanical contractions of the gastrointestinal smooth muscle. Slow waves generate the periodic activation of spontaneous inward currents (pacemaker currents). The aim of this study was to investigate the characterization of pacemaker currents of ICC. METHODS: The ICC in mice small intestine were cultured with stem cell factor for 2 days, and then we recorded pacemaker currents and slow waves using a whole-cell patch clamp technique. RESULTS: Under voltage clamp at -80 mV of holding potential, ICC generated pacemaker currents. Tetrodotoxin and nifedipine did not affect on the pacemaker currents. In addition, tetraethylammonium, 4-aminopyridine and glibenclamide did not affect on the pacemaker currents. The reduction of external Na+ concentrations inhibited pacemaker currents. Moreover, these currents were completely abolished in the external Ca2+-free condition. Gadolinium and flufenamic acid, inhibitors of non-selective cationic currents, inhibited pacemaker currents. Thapsigargin and cyclopiazoic acid, inhibitors of Ca2+-ATPase in endoplasmic reticulum, abolished pacemaker currents. Carbachol depolarized membrane potential and increased inward currents. CONCLUSIONS: These results suggest that pacemaker currents are mediated by the activation of non-selective cation channel and become a target of neurotransmitters in regulation of intestinal motility. PMID- 14532717 TI - [Clinical features and therapeutic responses of perianal lesions in Crohn's disease]. AB - BACKGROUND/AIMS: The aim of this study was to assess the clinical features and therapeutic responses of perianal lesions in Crohn's disease. METHODS: We retrospectively reviewed medical records of 94 cases with perianal lesions among 231 cases registered as Crohn's disease in Seoul National University Hospital between January 1975 and June 2001. The cases were investigated after dividing into two different groups. One group included the cases registered between January 1975 and December 1996 and the other group between January 1997 and June 2001. RESULTS: The perianal lesions were observed in 41% of the cases. Perianal lesions were more frequent in colonic Crohn's disease after 1997, but not before 1996. Fistula was significantly prevalent, followed by abscess, fissure, and stricture. Perianal lesions preceded intestinal symptoms such as abdominal pain and diarrhea in 25% of the cases. The remission rate with non-specific conservative treatment (10%) was lower than that before 1996. The remission rate after specific treatment (77%) was higher than that after surgery (53%). In cases of specific treatment, cumulative relapse rates were 7% in 1 year and 39% in 5 years. CONCLUSIONS: Crohn's disease should be considered in young male patients who have the refractory perianal lesions. Perianal lesions should be managed initially with medical treatment. PMID- 14532719 TI - [Role of hepatic stellate cells in the angiogenesis of hepatoma]. AB - BACKGROUND/AIMS: Hepatic stellate cells (HSCs) in the hepatocellular carcinoma are responsible for tumor encapsulation as a host defense mechanism. Recently, it was suggested that HSCs might play an important role in hepatic angiogenesis. Thus, HSCs in the HCC may be involved in tumor angiogenesis and pathogenesis of hepatic carcinogenesis. The purpose of this study was to examine the involvement of activated HSCs in the angiogenesis of hepatoma. METHODS: We investigated the effect of human HSC conditioned medium (CM) on the endothelial cell proliferation with or without stimulation of HepG2 CM, using [3H] thymidine incorporation assay. Additionally, we investigated the effect of HepG2 CM on HSCs proliferation and messenger RNA (mRNA) expression of various pro-angiogenic factors such as interleukin 8 (IL-8), basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) in HSCs. RESULTS: HSC CM caused a significant increase in DNA synthesis in human umbilical vein endothelial cells (HUVEC). The endothelial proliferation effect of HSCs was augmented by HepG2 CM. HepG2 CM significantly increased HSCs proliferation and stimulated IL-8 and bFGF mRNA expression in HSCs. CONCLUSIONS: HSCs promote endothelial proliferation through various soluble factors. The soluble factors secreted in HepG2 stimulate HSC proliferation and up-regulate mRNA expression of proangiogenic factors. This result suggests that HSCs may play an important role in the angiogenesis of hepatoma. PMID- 14532718 TI - [Inhibitory effect of angiotensin II receptor antagonist on the contraction and growth of hepatic stellate cells]. AB - BACKGROUND/AIMS: This study aimed to investigate the effects of angiotensin II (ANG II) and its receptor antagonist (losartan) on the contraction and growth of HSCs. METHODS: HSCs were isolated from Sprague Dawley rat and cultured at various conditions as follows: control, pretreatment of 10(-5) M ANG II, pretreatment of 10(-5) M endothelin, and pretreatment of 10(-5) M ANG II and 10(-6) M losartan. We conducted morphologic analysis with cellular area and length by image analysis system to estimate cell growth in each group. In addition, we measured the change of intracellular calcium currents via electrophysiological methods to evaluate the contractile effect of ANG II and losartan on HSCs. RESULTS: At the fifth day of incubation, the mean cellular area of ANG II-pretreated group and ANG II with losartan-pretreated group were 704.68+/-22.6 micro m2 and 332.90+/-32.6 micro m2, respectively. This difference was statistically significant (p<0.05). ANG II induced an increase in the intracellular calcium current by 22.0+/-3.0% compared with basal current level (p<0.05). However, when losartan was pretreated, ANG II did not cause a significant increase in calcium current (3.1+/-0.8%, p>0.05). CONCLUSION: ANG II accelerates the contraction and growth of HSCs, while its receptor blocker, losartan, inhibits the contraction and growth of HSCs. PMID- 14532720 TI - [The effect of endotoxin on the expression of tissue factor in the CCl4-treated rat liver]. AB - BACKGROUND/AIMS: Endotoxemia is known to cause hepatic microcirculation disturbance and hepatic injury through blood coagulation and thrombosis in the cirrhotic patients. This study was to investigate the effect of endotoxin on the expression of tissue factor in rat liver with persistent injury. METHODS: Male Sprague-Dawley rats were used for this experiment. Persistent hepatic injury was induced by injecting 0.1 mL of CCl4 and 0.1 mL of mineral oil per 100 grams of body weight, intraperitoneally. Twenty-four rats were divided into 4 groups and lipopolysaccharide (LPS, E. coli O111:B4) was injected intraperitoneally in 3 groups in a dose of 200 microgram/kg, 400 microgram/kg, and 800 microgram/kg, respectively. Expression of tissue factor was evaluated by real time PCR method. SYBR Green I, a DNA binding fluorophore was used for the detection of PCR product. RESULTS: Expression of hepatic tissue factor tended to increase according to the amounts of LPS. Specially, the expression of tissue factor was significantly increased in the group into which 400 microgram/kg of LPS was injected, compared with control group (p<0.05). CONCLUSIONS: Expression of tissue factor tends to increase according to the amounts of LPS in the persistent hepatic injury. LPS may play an important role in the expression of tissue factor in the CCl4-treated rat liver. PMID- 14532721 TI - [A result of surgical treatment for ductal adenocarcinoma of the head of the pancreas]. AB - BACKGROUND/AIMS: Ductal adenocarcinoma of the head of the pancreas can only be cured with pancreaticoduodenectomy. However, the prognosis after resection has been poor. This study analyzed the possible prognostic factors of resected ductal adenocarcinoma through pancreaticoduodenectomy in a single institution. METHODS: Between Oct. 1994 and May. 2002, 81 patients underwent pancreatoduodenectomy for ductal adenocarcinoma of the head of the pancreas. Perioperative and pathologic factors were analyzed with univariate and multivariate analysis. RESULTS: The overall 5-year survival rate of the 81 patients was 10.8% with median survival of 11.8 months. Ninety-six percent of the patients were in stage IIa, IIb or III. In univariate analysis, factors favoring survival were as follows: CA19-9 antigen0.70). Furthermore, intraclass correlation for the first and second administration of the VCI was 0.79. In addition, there was good correlation between the validated SF-36 and FACT-VCI (r = 0.81). The result was initial validation of a questionnaire for the assessment of HRQOL in patients following RC and UD. CONCLUSIONS: Instruments designed to measure accurately HRQOL following RC and UD are in early development. We have now constructed and validated a disease and treatment specific questionnaire that can objectively assess HRQOL following RC and UD. A prospective longitudinal study of FACT-VCI is currently ongoing. PMID- 14532810 TI - Fear of cancer recurrence in patients undergoing definitive treatment for prostate cancer: results from CaPSURE. AB - PURPOSE: Contemporary cancer treatments have resulted in patients living longer but with the risk of disease recurrence. Studies suggest that fear of recurrence is a significant burden. We described fear of cancer recurrence in patients with prostate cancer undergoing treatment with radical prostatectomy (RP), radiation (XRT) or brachytherapy (BT). MATERIALS AND METHODS: A total of 519 patients (326 RP, 53 XRT, 140 BT) were identified from CaPSURE (Cancer of the Prostate Strategic Urologic Research Endeavor), a national longitudinal registry of men with prostate cancer. To be included in the study patients had to complete at least 1 pretreatment and 2 posttreatment health related quality of life questionnaires and have complete clinical information. Fear of cancer recurrence was assessed with a validated 5-item scale, and was described at baseline and up to 2 years after treatment. Multivariate linear regression was performed to determine significant predictors of fear of cancer recurrence. RESULTS: Men receiving XRT were older and had worse clinical disease characteristics than patients treated with RP or BT. For all groups fear of cancer recurrence was more severe before treatment and improved after treatment but did not change substantially in the 2 years thereafter. Regression revealed that only general health and mental health were important predictors of fear of cancer recurrence. No other general or disease specific health related quality of life domains or clinical characteristics contributed appreciable explanatory power. CONCLUSIONS: Fear of prostate cancer recurrence imposes a substantial burden in patients before and after treatment. Understanding the fear of cancer recurrence associated with different treatments can help physicians better counsel patients and promote psychological well-being. PMID- 14532811 TI - Intravenous stent placement for treatment of the nutcracker syndrome. PMID- 14532812 TI - Subcutaneous emphysema, pneumomediastinum and bilateral pneumothoraces after laparoscopic pyeloplasty. PMID- 14532813 TI - Laparoscopic management of chylous ascites after donor nephrectomy. PMID- 14532814 TI - Chronic pain syndrome after laparoscopic radical nephrectomy. PMID- 14532815 TI - Spontaneous rupture of adrenal carcinoma during the puerperal period. PMID- 14532816 TI - Thrombosed urethral hemangioma. PMID- 14532817 TI - Adenofibroma of the testis discovered at exploration for torsion after trauma. PMID- 14532818 TI - Phyllodes tumor of the prostate: a rare condition requiring careful surveillance. PMID- 14532820 TI - Bilharzial pyelitis: a rare cause of secondary ureteropelvic junction obstruction. PMID- 14532819 TI - Left perinephric abscess caused by Salmonella enteritidis due to colon perforation. PMID- 14532821 TI - Transcutaneous laser hair ablation for management of intraurethral hair after hypospadias repair: initial experience. PMID- 14532822 TI - Encapsulation of a porcine dermis pubovaginal sling. PMID- 14532823 TI - Benign prostatic stromal hyperplasia with bizarre nuclei. PMID- 14532824 TI - Novel technique in the management of low flow priapism. PMID- 14532825 TI - Recurrent polyarteritis nodosa limited to the testis. PMID- 14532826 TI - Re: Enhancement of diethylstilbestrol induced cytotoxicity by bcl-2 antisense oligodeoxynucleotides and a glutathione depletor for prostate cancer. PMID- 14532827 TI - Re: Charlson co-morbidity index as a predictor of outcome after surgery for renal cell carcinoma with renal vein, vena cava or right atrium extension. PMID- 14532828 TI - Re: Disease progression and survival of patients with positive lymph nodes after radical prostatectomy. Is there a chance of cure? PMID- 14532829 TI - Re: Sacral neuromodulation for the symptomatic treatment of refractory interstitial cystitis: a prospective study. PMID- 14532830 TI - Dried fruits decrease nocturnal polyuria. PMID- 14532831 TI - Re: Influence of voiding dysfunction on the outcome of endoscopic treatment for vesicoureteral reflux. PMID- 14532832 TI - Renal size and function in patients with neuropathic bladder due to myelomeningocele: the role of growth hormone. AB - PURPOSE: Patients with spina bifida have smaller kidneys than healthy individuals. We evaluated the correlation between small size and decreased renal function, and the possible role of growth hormone deficiency. MATERIALS AND METHODS: A total of 54 patients (mean age 11.5 years, median 11, standard deviation +/- 4.52) were healthy except for neuropathic bladder due to spina bifida. Renal function was evaluated with mercaptoacetyltriglycine renal scintigraphy and creatinine clearance. Renal anatomy was evaluated with renal ultrasound and voiding cystourethrography. Serum insulin-like growth factor-1 (IGF-1) levels were measured in all patients with immunoradiometric assay. Renal measurements in our patients were compared using the Sutherland nomogram. RESULTS: A total of 22 patients (41%) had smaller kidneys than normal subjects and 31 appeared to have creatinine clearance values lower than 120 ml per minute per 1.73 m2. The statistical comparison between kidney size and creatinine clearance was significant (p <0.05, r = 0.381). Scintigraphic data showed total effective renal plasma flow less than 568 ml per minute per 1.73 m2 body surface area (normal mean value for age). Comparison between effective renal plasma flow and creatinine clearance was significant (p <0.05, r = 0.31). Serum levels of IGF 1 were normal for age in all patients (mean 332.06 ng/ml, median 303.4, range 39.4 to 732.3). CONCLUSIONS: The kidneys are smaller in patients with spina bifida than in healthy subjects when compared using the Sutherland nomogram. There is a significant correlation between smaller renal length and decreased renal function in all patients, even in those who are healthy except for neurogenic bladder secondary to spina bifida. IGF-1 levels were normal for age, and, therefore, these patients had no growth hormone deficiency. These findings call into question the hypothesis that growth hormone deficiency contributes to smaller kidney size. Other hypotheses can be suggested, such as a defect of embryological growth secondary to malformation, or the result of a defect in homocysteine-methionine metabolism. PMID- 14532833 TI - Bilateral laparoscopic transperitoneal nephrectomy with early peritoneal dialysis in an infant with the nephrotic syndrome. PMID- 14532834 TI - Results of complete penile disassembly for epispadias repair in 42 patients. AB - PURPOSE: We evaluated the Mitchell complete penile disassembly technique for epispadias repair. MATERIALS AND METHODS: A total of 42 males 1 month to 22 years old presented for repair of epispadias between 1998 and 2002. Cases were divided into 2 groups. Group 1 included 29 cases of complete epispadias as a component of bladder exstrophy, 8 with previous continent urinary diversion. Group 2 included 13 cases of epispadias alone (10 primary and 3 secondary). Of the 29 patients in group 1, 21 underwent complete penile disassembly as part of 1-stage primary closure of bladder exstrophy. RESULTS: Mean followup was 37.5 months (range 6 to 52). Ischemic changes at the glans penis were observed in 5 cases during our initial experience. Ventral orthotopic meatus was observed in all 42 patients, conical glans in 40 (95.2%), straight shaft in 34 (81%) and urethral fistula in 1 (2.4%). There were no cases of dehiscence, meatal stenosis or urethral stricture. Erectile function was preserved in all patients. CONCLUSIONS: Complete penile disassembly is a safe procedure that can provide normalization of the urethra and penis together with satisfactory cosmetic and functional outcome. PMID- 14532835 TI - Dynamic renal scintigraphy in children with vesicoureteral reflux and suspected coexisting ureteropelvic junction obstruction. AB - PURPOSE: We evaluated whether findings on voiding cystourethrography suggesting ureteropelvic junction (UPJ) obstruction coexists with vesicoureteral reflux (VUR) are associated with parameters on dynamic renal scintigraphy that support significant obstruction. MATERIALS AND METHODS: We reviewed records of 44 patients referred for scintigraphy after voiding cystourethrography performed at age 1 day to 9.4 years (mean 7 months, median 1.7 months) showed VUR and findings suggestive of UPJ obstruction (blockage of contrast material at the UPJ, contrast dilution in the renal pelvis, slow renal pelvic drainage). Results were correlated with Society for Fetal Urology hydronephrosis grade and ureteral morphology. RESULTS: Halftime was in the obstructive range (20 minutes or greater) for 7 of 47 kidneys (15%). The prevalence of a post-furosemide pelvicaliceal drainage halftime in the obstructive range increased with hydronephrosis grade (0% grade 1, 17% grade 2, 50% grade 3 to 4, p = 0.002) but did not vary with ureteral morphology (p = 0.08). In 12 of 38 cases (31%) where suspected UPJ obstruction was unilateral and a contralateral kidney was present differential uptake of the affected kidney was less than 45%. The prevalence of differential uptake less than 45% was higher in patients with than without ureteral dilatation (48% vs 12%, p = 0.02) but did not vary with hydronephrosis grade (p = 0.93). CONCLUSIONS: In children with VUR and suspected coexisting UPJ obstruction dynamic renal scintigraphy may support significant obstruction when hydronephrosis is at least moderate in degree or ureteral dilatation is present but is unlikely to do so if neither is observed. PMID- 14532836 TI - Lower urinary tract dysfunction in children with generalized hypermobility of joints. AB - PURPOSE: We assessed the relationship between general joint hypermobility (GJH) and lower urinary tract symptoms presenting as nonneurogenic bladder sphincter dysfunction in children. MATERIALS AND METHODS: Parents of 89 children 5 to 12 years old who were diagnosed with GJH according to the Beighton and Bulbena scales were asked to fill out a questionnaire concerning symptoms of nonneurogenic bladder sphincter dysfunction, namely daytime urinary incontinence, nighttime urinary incontinence, urinary tract infection, constipation and fecal soiling. The control group comprised 116 healthy schoolchildren 5 to 12 years old. RESULTS: Constipation was reported in 19% of boys with GJH and 4% of male controls (p = 0.02). Fecal soiling occurred more often in the GJH group than in the control group (34% versus 18%, p = 0.07). In girls daytime and nighttime urinary incontinence was more prevalent in the GJH group (38% and 14%, respectively) than in controls (13% and 2%, respectively, p = 0.004 and p = 0.02, respectively). Of girls with GJH 24% had a history of urinary tract infections compared with 11% of the control group (p = 0.08). CONCLUSIONS In children with generalized hypermobility of joints symptoms of nonneurogenic bladder sphincter dysfunction are more prevalent. In boys this condition manifests as constipation and possibly fecal soiling, and in girls as urinary incontinence and possibly urinary tract infections. PMID- 14532837 TI - Bladder dysfunction after bilateral ectopic ureterocele repair. AB - PURPOSE: We examined postoperative outcome, with emphasis on bladder function, in pediatric patients who underwent bilateral ectopic ureterocele repair. MATERIAL AND METHODS: We reviewed the records of 117 patients with orthotopic and ectopic ureteroceles treated between 1977 and 2000. Twelve of these patients had bilateral ectopic ureteroceles. All patients with bilateral ureteroceles were females 1 day to 2 years old at referral. Initial treatment was transureteral puncture in 6 cases, transurethral unroofing in 2 and extravesical bilateral reimplantation in 1. Of the remaining 3 patients the initial treatment was unilateral heminephroureterectomy in 1 and bilateral heminephroureterectomy in 2. Definitive treatment included bilateral upper to lower ureteroureterostomy, ureterocelectomy with trigonal and bladder neck reconstruction, and bilateral ureteroneocystotomy with or without tapering of the recipient ureter. In patients who underwent upper pole partial nephrectomy the distal upper pole ureter was removed in conjunction with trigonal surgery and ureteroneocystotomy. RESULTS: There was significant morbidity in this group of patients, including voiding dysfunction and poor bladder emptying with residual urine greater than 20% of bladder capacity in 7 of 10 patients studied by serial bladder ultrasonography. Urodynamic evaluation in 3 patients revealed increased bladder compliance and large volume residual urine. Of these 3 patients 2 perform clean intermittent catheterization. Recurrent symptomatic bacteruria was noted in 7 of the 12 patients. CONCLUSIONS: Patients undergoing bilateral ectopic ureterocele repair are at increased risk for postoperative voiding dysfunction. Whether this risk is present preoperatively or is a result of trigonal surgery is unclear. PMID- 14532838 TI - Trospium chloride for the treatment of detrusor instability in children. AB - PURPOSE: We assessed the efficacy and most appropriate dosage of trospium chloride (TCl) for managing bladder instability in children as compared with a placebo. MATERIALS AND METHODS: A total of 58 patients with bladder instability were allocated at random to 1 of 5 groups-10, 15, 20 or 25 mg TCl, or placebo administered daily in a multicenter, randomized, single-blind clinical study. Patients were treated for 21 days, and current symptoms, voiding diary and urodynamic values were collected at the beginning and end of the treatment period. All adverse events were recorded at the last visit. RESULTS: Of 50 patients treated with TCl 41 (82%) had a positive therapeutic result (excellent, good or fair) versus only 3 of 8 patients with improvement in the placebo group (37.5%, p = 0.006). In all responding patients clinical symptoms either resolved or decreased markedly, and in 37 (74%) this improvement was accompanied by urodynamic improvement. In these 37 children the average number of uninhibited contractions decreased by 54.3% (p <0.0001) and the volume at first contraction increased by 71.4% (p = 0.001). There were no statistically significant differences with regard to therapeutic efficacy between TCl dosages. Fourteen patients (9 with TCl, 5 with placebo) showed no clinical improvement, although some had improved urodynamic parameters. Furthermore, TCl was well tolerated with few patients (10%) experiencing adverse effects. CONCLUSIONS: Trospium chloride (10 to 25 mg total daily dosage, split into 2 doses) is an effective option for the management of detrusor instability in children. PMID- 14532839 TI - The role of Ras superfamily proteins in bladder cancer progression. AB - PURPOSE: The prognosis of patients with bladder cancer is strongly dependent on whether the lesion is superficial or invasive at initial presentation. In addition, a significant fraction of patients presenting with superficial disease have invasive tumor during followup. Understanding how superficial bladder cancer progresses to invasive forms of the disease is of paramount importance for early diagnosis and successful treatment. Molecular mechanisms underlying bladder cancer progression are being elucidated. We reviewed the roles that members of the Ras superfamily of monomeric G proteins, an important class of cellular regulator, have in bladder cancer and its progression. MATERIALS AND METHODS: We performed MEDLINE searches focusing on members of the Ras superfamily of monomeric G proteins and their involvement in transitional cell carcinoma, which is the most common form of bladder cancer. General involvement in cancer of key superfamily members, focusing on mechanisms and downstream pathways, was also reviewed through MEDLINE and manual bibliographic searches. RESULTS: With more than 100 members in humans the Ras superfamily is a diverse group of monomeric G proteins. These proteins regulate many cellular processes, such as cell cycle progression, actin cytoskeletal dynamics and membrane traffic. Members of the Ras and Rho family are also known to be involved in human cancer through mutation, over expression and dysregulation. In this review we focus on bladder cancer. In particular we focus on how H-Ras, RalA/B and RhoGDI2, a regulator of Rho family members, participate in bladder cancer progression and how their participation may be related to other molecules associated with bladder cancer progression, such as epidermal growth factor receptor, p53 and PTEN (phosphatase and tensin homologue deleted on chromosome 10). CONCLUSIONS: The findings discussed offer the hopeful possibility that signaling pathways mediated by Ras superfamily members may offer new opportunities for diagnostic and therapeutic interventions in bladder cancer. PMID- 14532840 TI - Expression of lipoxygenase in human bladder carcinoma and growth inhibition by its inhibitors. AB - PURPOSE: The metabolism of arachidonic acid by the cyclooxygenase or lipoxygenase pathway generates eicosanoids, which have been implicated in the pathogenesis of various human diseases, including cancer. They are now believed to have important roles in tumor promotion, progression and metastasis. The involvement of lipoxygenase expression and function in tumor growth and metastasis has been reported in human tumor cell lines. MATERIALS AND METHODS: The expression of 5 and 12-lipoxygenase in patients with bladder tumor and chronic cystitis, and in normal bladder tissues was examined. We also examined the effects of their inhibitors on cell proliferation in a bladder cancer cell line. The expression of 5 and 12-lipoxygenase protein was detected by immunohistochemistry. The effects of lipoxygenase inhibitors on bladder cancer cell growth were examined by MTT (3 [4,5-dimethylthiazol-2-thiazolyl]-2,5-diphenyltetrazolium bromide) assay, while Hoechst (Sigma Chemical Co., St. Louis, Missouri) staining was used to determine whether lipoxygenase inhibitors induce apoptosis. RESULTS: While slight 5 and 12 lipoxygenase expression was detected in chronic cystitis and normal bladder tissues, marked 5 and 12-lipoxygenase expression was detected in bladder cancer tissues. Lipoxygenase inhibitors caused marked inhibition of bladder cancer cells in a concentration and time dependent manner. Cells treated with lipoxygenase inhibitors showed chromatin condensation, cellular shrinkage, small membrane bound bodies (apoptotic bodies) and cytoplasmic condensation. CONCLUSIONS: Lipoxygenase is induced in bladder cancer. Results suggest that lipoxygenase inhibitors may mediate potent antiproliferative effects against bladder cancer cells. Thus, lipoxygenase may become a new target in the treatment of bladder tumors. PMID- 14532841 TI - The anti-angiogenic activity of human endostatin inhibits bladder cancer growth and its mechanism. AB - PURPOSE: We investigated whether recombinant human endostatin can inhibit the growth of bladder cancer in an experimental model and its possible mechanism of action. MATERIALS AND METHODS: The recombinant human endostatin protein was induced and confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot assays. Its biological activities and the possible mechanisms of action were studied in vitro and in vivo. RESULTS: Recombinant human endostatin inhibited the proliferation of endothelial cells (ECV304) but not bladder tumor cells (EJ). Endostatin induced the expression of matrix metalloproteinases and tissue inhibitors of matrix metalloproteinases in bladder cancer cells. Endostatin slowed the growth of xenograft bladder tumors. Immunohistochemistry revealed that endostatin blocked angiogenesis by decreasing vascular endothelial growth factor expression and inducing apoptosis in bladder cancer cells. CONCLUSIONS: These findings demonstrate that endostatin can inhibit xenograft bladder cancer growth and this effect is likely to be mediated by regulating matrix metalloproteinases, tissue inhibitors of matrix metalloproteinases and vascular endothelial growth factor expression, and by inducing apoptosis. PMID- 14532842 TI - Cytokine gene expression in a mouse model: the first instillations with viable bacillus Calmette-Guerin determine the succeeding Th1 response. AB - PURPOSE: Bacillus Calmette-Guerin (BCG) therapy for superficial bladder cancer is immune dependent and activation of a Th1 immune response is probably required for clinical efficacy. Given the empirical approach to improving BCG therapy we investigated in a mouse model the consequences of modifications in BCG therapy with regard to Th1 and Th2 cytokine responses in the bladder. These studies may provide a rationale for possible modifications of the established clinical treatment protocol. MATERIALS AND METHODS: The dynamics of Th1 (interferon-gamma, interleukin [IL]-2, IL-12p40 and tumor necrosis factor-alpha) and Th2 (IL-10 and IL-4) cytokine responses during and after 6 once weekly intravesical BCG instillations in mice was determined by a semiquantitative reverse transcriptase polymerase chain reaction based method. RESULTS: During 6 weekly BCG instillations a dose and time dependent induction of the various Th1 as well as Th2 cytokines was observed. The response pattern was comparable to urinary cytokine induction patterns in patients. Electrocauterization prior to BCG instillations led to lower and more variable levels of cytokine polymerase chain reaction products compared with noncauterization. Lowering the dose of BCG seemed to affect the Th1 cytokine response most, whereas the Th2 response was less influenced by dilution of the BCG preparation. Six instillations with nonviable BCG induced Th2 but failed to induce Th1 cytokines, which may explain the necessity of BCG viability for antitumor activity. However, when mice were first treated 3 times with viable BCG and subsequently received 3 instillations with killed BCG, the Th1 and Th2 cytokine pattern was comparable to the standard 6 week regimen with viable BCG. CONCLUSIONS: The model seems an appropriate one in which to investigate changes in Th1 and Th2 cytokine gene expression levels in bladders resulting from modifications in intravesical BCG treatment. It was possible to induce a local Th1 cytokine response with nonviable BCG provided that local sensitization to BCG antigens had occurred during preceding instillations with a viable BCG preparation. Whether such an approach could decrease BCG therapy toxicity, while maintaining antitumor efficacy, remains to be further investigated in patients. PMID- 14532843 TI - Androgen dependent regulation of bacillus Calmette-Guerin induced interleukin-6 expression in human transitional carcinoma cell lines. AB - PURPOSE: Autocrine expression of interleukin (IL)-6 by transitional cell carcinoma (TCC) in response to bacillus Calmette-Guerin (BCG) may have an important role in promoting BCG adherence to TCC and consequently in BCG treatment efficacy. IL-6 expression in response to BCG requires nuclear factor (NF)-kappaB mediated signal transduction. We evaluated the influence of androgens on BCG induced, NF-kappaB mediated IL-6 expression. MATERIALS AND METHODS: Reverse transcriptase-polymerase chain reaction was used to confirm androgen receptor expression in the human TCC lines 253J and T24. A reporter construct containing an androgen response element was used to establish the integrity of androgen mediated signal transduction. Subsequently the dose dependent effect of dihydrotestosterone (DHT) on BCG induced IL-6 expression and NF-kappaB signaling was evaluated. Two pharmacological androgen receptor blockers were used to determine if receptor blockade inhibited the effect of DHT on activation of the androgen response element, NF-kappaB signaling and BCG induced IL-6 expression. RESULTS: The 2 human TCC lines expressed androgen receptor and demonstrated intact androgen stimulated signaling pathways. DHT suppressed BCG induced, NF kappaB mediated signaling and IL-6 expression in a dose dependent manner. DHT decreased mRNA levels of IL-6, expression of the full-length IL-6 promoter construct and expression of an NF-kappaB specific reporter construct in response to BCG relative to controls. Competitive pharmacological blockade of androgen receptor inhibited the effect of DHT on BCG induced signaling in dose dependent fashion. CONCLUSIONS: DHT down-regulates NF-kappaB mediated IL-6 expression by human TCC lines in response to BCG. This effect depends on a functional androgen receptor signaling pathway and it can be blocked by the inhibition of androgen/androgen receptor binding. PMID- 14532844 TI - Progesterone induces the proliferation of urothelial cells in an epidermal growth factor dependent manner. AB - PURPOSE: We have previously reported that estrogen induced proliferation of urothelial cells is modulated by nerve growth factor (NGF). In this study we investigated whether progesterone induces urothelial cell proliferation and whether this effect is modulated by NGF or by epidermal growth factor (EGF). MATERIALS AND METHODS: Experiments were performed using human urothelial cells immortalized by human papillomavirus E6. Cell proliferation was determined using the alamarBlue (Trek Diagnostic, Westlake, New York) assay. Human papillomavirus were seeded in 48-well plates. They were incubated with 5% alamarBlue and different concentrations of progesterone, EGF or NGF in the presence or absence of neutralizing EGF or NGF antibody, K252a (an inhibitor of trkA, the high affinity receptor for NGF), Ru-486 (an antagonist of progesterone and glucocorticoid receptor) or ZK 137 316 (a specific antagonist of progesterone receptor). Immunoblotting was performed using specific antibodies for progesterone receptor, glucocorticoid receptor or EGF receptor. EGF content in conditioned medium was determined by enzyme-linked immunosorbent assay. RESULTS: In the presence of 10 nM to 1 microM progesterone urothelial cell proliferation was significantly increased 8.6% to 51.1%. This effect was abolished by ZK137 316 or by Ru-486. Hydrocortisone also induced urothelial cell proliferation. This effect was blocked by Ru-486 but not by ZK137 316. In addition, progesterone stimulated urothelial cell proliferation was inhibited by neutralizing EGF antibody but not by NGF antiserum or K252a. We also found that EGF synthesis and release by urothelial cells was increased by exogenous progesterone. This effect of progesterone was inhibited by ZK 137 316. CONCLUSIONS: These findings indicate that progesterone has the capacity to induce urothelial cell proliferation through its cognate receptor and this effect is mediated by EGF but not by NGF. PMID- 14532845 TI - 5alpha-reductase type 1 immunostaining is enhanced in some prostate cancers compared with benign prostatic hyperplasia epithelium. AB - PURPOSE: In the prostate testosterone is converted to the more potent androgen dihydrotestosterone by the enzymes 5alpha-reductase (5alphaR) types 1 (5alphaR1) and 2 (5alphaR2). Since 5alphaR2 is the dominant prostatic enzyme, the 5alphaR2 selective inhibitor finasteride has been widely used to treat benign prostatic hyperplasia (BPH). However, inhibition of both 5alphaR enzymes provides a greater decrease in serum dihydrotestosterone. We developed a specific antibody to 5alphaR1 and assessed expression in BPH and prostate cancer (pCa) tissue. The presence of this isoenzyme in localized prostate cancer would provide a rationale for assessing the efficacy of dual inhibition for prostate cancer prevention. MATERIALS AND METHODS: A polyclonal antibody to 5alphaR1 was developed and validated using 5alphaR1 and 5alphaR2 transfected COS-1 cells. A total of 26 BPH and 53 pCa specimens were assessed for 5alphaR1 protein expression using immunocytochemical methods. Also, 29 BPH and 37 pCa specimens were assayed for 5alphaR1 and 5alphaR2 enzyme activity. RESULTS: Specificity of the 5alphaR1 antibody was confirmed using transfected COS-1 cells. Cells transfected with 5alphaR1 showed specific staining in immunocytochemistry experiments and on Western blotting of cell lysates the expected 24 kDa band was observed. High intensity immunoreactivity for 5alphaR1 was observed in the tumor epithelium of 28% of pCa specimens. No high intensity epithelial staining was observed in BPH specimens. In 19% of pCa and 7% of BPH specimens 5alphaR1 enzyme activity was detected. CONCLUSIONS: The presence of increased 5alphaR1 in some prostatic malignancies suggests that it is worthwhile to investigate the use of a dual 5alphaR inhibitor to prevent or treat early stage prostate cancer. PMID- 14532846 TI - Inhibition of dendropoiesis by tumor derived and purified prostate specific antigen. AB - PURPOSE: Prostate specific antigen (PSA) is a serine protease produced by the prostate gland at high concentrations. Serum PSA may be significantly elevated in prostate cancer and benign prostatic diseases. It has recently become evident that, in addition to being a tissue and/or serum marker, PSA may also have biological effects. Despite the voluminous literature on this biomarker in the diagnosis of prostatic diseases relatively few reports have addressed the issue of the physiological function, biological role and immune effects of PSA in the context of prostate cancer development and progression. MATERIALS AND METHODS: Human dendritic cell (DC) cultures were generated from CD34+ hematopoietic precursors in the presence of PSA. The DC phenotype was assessed by flow cytometry and DC ability to induce T-cell proliferation was detected by allogeneic mixed lymphocyte reaction assay. DCs were also generated in co cultures with LNCaP cells in the presence of antiPSA antibodies. The concentrations of PSA in cultures were determined by the AXSYM System (Abbott Laboratories, Wiesbaden, Germany). RESULTS: We noted that purified and LNCaP derived PSA inhibited the generation and maturation of DC (dendropoiesis) in vitro, which might have a crucial role in the induction and regulation of specific antitumor immune responses. The addition of active PSA to DC cultures significantly inhibited the generation and maturation of DC, as assessed by the levels of expression of CD83, CD80, CD86 and HLA DR. The ability of DC to induce T-cell proliferation, which depends on the expression of co-stimulatory and major histocompatibility complex molecules, was also suppressed in PSA treated DC cultures. CONCLUSIONS: The antidendropoietic effect of PSA in vitro suggests a new mechanism of prostate cancer induced immunosuppression and tumor escape, and provides novel evidence of the immunoregulatory properties of PSA. PMID- 14532847 TI - Modulation of prostate carcinoma cell growth and apoptosis by chromogranin A. AB - PURPOSE: We elucidated the effect and possible pathways of chromogranin A in regulating prostatic cancer cell growth. MATERIALS AND METHODS: Chromogranin A expression in prostatic cancer cell lines were detected with immunofluorescence flow cytometry (FCM) and the inhibition of cell growth due to chromogranin A antibody was measured using microculture tetrazolium. Cell cycle and RNA changes were evaluated with acridine orange FCM. Intracellular chromogranin A variations were detected with FCM, as were apoptotic changes, p53, Fas and tumor necrosis factor-alpha. Apoptosis and caspase-3 expression of tumor cells was assessed with dual immunohistochemical staining. RESULTS: Increased chromogranin A expression was observed in PC-3, DU145 and LNCap cells independent of hormone dependence. Dose and time dependent growth inhibition occurred at 12 hours. Chromogranin A antibody arrested PC3 cells in the S-phase immediately after treatment. The number of G0/G1 and G2/M cells subsequently decreased. PC3 tumor cells had transiently increased RNA at 12 hours with a marked decrease at 48 hours. Decreasing chromogranin A expression started at 12 hours and was most prominent at 48 hours. Apoptotic cells markedly increased at 12 hours with an increase in p53, Fas and tumor necrosis factor-alpha (Fas more than the others). Increased apoptotic cell and caspase-3 expression was observed on immunohistochemical stains. CONCLUSIONS: Chromogranin A is an important neuropeptide regulating the growth of prostate cancer cells. Specific antibodies can suppress its function through apoptotic pathways (Fas and caspase-3), leading to programmed cell death. Chromogranin A antibody mediated apoptosis may be a specific alternative treatment for prostate cancer. PMID- 14532848 TI - Cyclooxygenase-2 inhibitor NS398 enhances antitumor effect of irradiation on hormone refractory human prostate carcinoma cells. AB - PURPOSE: We examined the potential therapeutic effect of NS398, a selective cyclooxygenase-2 (COX-2) inhibitor, combined with irradiation on human prostate adenocarcinoma DU145 cells. MATERIALS AND METHODS: The effect on tumor growth, proliferation testing and clonogenic survival was determined to evaluate its antitumor effect when exposed to NS398 or combined with irradiation. Immunoblotting analyses were done to detect the expression of COX-2, nuclear factor-kappaB p50 and Rel A p65 because evidence suggested a biological association of COX-2 with alterations in these markers. Reverse transcriptase polymerase chain reaction was also performed to show its effect on the transcription level of COX-2. RESULTS: Exposure of DU145 cells to NS398 alone suppressed proliferation in a dose and time dependent manner. Examination of the NS398 effect on the radiation response showed marked enhancement of radiosensitivity. Western blot indicated that NS398 down-regulated the expression of COX-2, nuclear factor-kappaB, p50 and Rel A p65, whereas the effect was more pronounced when combined with irradiation. Reverse transcriptase-polymerase chain reaction showed that the NS398 antitumor effect was associated with COX-2 transcription inhibition. Importantly COX-2 expression was enhanced by irradiation but this phenomenon was abolished when cells were exposed to NS398. Inhibition of tumor growth in animal model was observed when mice were treated with NS398 alone and irradiation alone, and this effect was maximal when treated with NS398 and irradiation. CONCLUSIONS: These results suggest that NS398 could be used as a potential therapeutic agent combined with irradiation for prostate adenocarcinoma. PMID- 14532849 TI - Laminin expression patterns in human ureteral tissue. AB - PURPOSE: Laminins are extracellular matrix proteins that are involved in various cellular functions, including adhesion, proliferation and differentiation. In this study we examined the expression patterns of the laminin chains in human ureteral tissue in vitro and in vivo. MATERIALS AND METHODS: We screened the expression of laminin chains at the mRNA level and determined the major laminins expressed in epithelial and stromal cells of human ureteral tissue by reverse transcriptase-polymerase chain reaction and Western blot analysis. We also examined their expression in vivo by immunofluorescence study. RESULTS: Reverse transcriptase-polymerase chain reaction analysis revealed that urothelial cells in vitro expressed laminins alpha1, alpha3, alpha5, beta1, beta2, beta3, gamma1 and gamma2, while stromal cells expressed alpha1, alpha2, alpha4, alpha5, beta1, beta2 and gamma1. Western blot analysis under reduced conditions confirmed that urothelial cells expressed laminin alpha3 and alpha5, while stromal cells strongly expressed alpha4 and alpha2. Immunofluorescence labeling with antilaminin alpha chain antibodies in human ureteral tissues confirmed that laminin alpha2, alpha3 and alpha5 chains were strongly expressed in the basement membrane of the urothelium, while alpha4 was expressed predominantly in smooth muscle cells. CONCLUSIONS: The most prominent laminin of urothelial cells was laminin 5 (alpha3beta3gamma2), whereas in stromal cells we noted laminins 8/9 (alpha4beta1/2gamma1). Laminin alpha5 showed the widest distribution pattern in vivo. Our data may be useful for understanding the mechanism involved not only in cell growth and differentiation, but also in cancer invasion in the urinary tract. PMID- 14532850 TI - Gene therapy for bladder pain with gene gun particle encoding pro opiomelanocortin cDNA. AB - PURPOSE: Interstitial cystitis is a bladder hypersensitivity disease associated with bladder pain that has been a major challenge to understand and treat. We hypothesized that targeted and localized expression of endogenous opioid peptide in the bladder could be useful for the treatment of bladder pain. Pro opiomelanocortin (POMC) is one of such precursor molecules. In this study we developed a gene gun method for the transfer of POMC cDNA in vivo and investigated its therapeutic effect on acetic acid induced bladder hyperactivity in rats. MATERIALS AND METHODS: Human POMC cDNA was cloned into a modified pCMV plasmid and delivered into the bladder wall of adult female rats by direct injection or the gene gun. Three days after gene therapy continuous cystometrograms were performed using urethane anesthesia by filling the bladder (0.08 ml per minute) with saline, followed by 0.3% acetic acid. Bladder immunohistochemical testing was used to detect endorphin after POMC cDNA transfer. RESULTS: The intercontraction interval was decreased after intravesical instillation of acetic acid (73.1% or 68.1% decrease) in 2 control groups treated with saline or the gene gun without POMC cDNA, respectively. However, rats that received POMC cDNA via the gene gun showed a significantly decreased response (intercontraction interval 35% decreased) to acetic acid instillation, whereas this antinociceptive effect was not detected in the plasmid POMC cDNA direct injection group. This effect induced by POMC gene gun treatment was reversed by intramuscular naloxone (1 mg/kg), an opioid antagonist. Increased endorphin immunoreactivity with anti-endorphin antibodies was observed in the bladder of gene gun treated animals. CONCLUSIONS: The POMC gene can be transferred in the bladder using the gene gun and increased bladder expression of endorphin can suppress nociceptive responses induced by bladder irritation. Thus, POMC gene gun delivery may be useful for the treatment of interstitial cystitis and other types of visceral pain. PMID- 14532851 TI - The use of halofuginone in limiting urethral stricture formation and recurrence: an experimental study in rabbits. AB - PURPOSE: We developed a reproducible animal model for the induction of urethral stricture in the rabbit and evaluated the role of halofuginone in limiting stricture formation. MATERIALS AND METHODS: A total of 20 New Zealand male rabbits were used in the first phase of the experiment. Bulbar urethral stricture was induced by electrocoagulation. The animals were then randomly assigned to 2 groups of 10 each, which received a diet containing halofuginone or a normal diet. In the second phase electrocoagulation induced stricture was treated with visual internal urethrotomy in 45 rabbits. These rabbits were randomly assigned to 2 groups, namely a halofuginone and a control group. RESULTS: In the first phase stricture developed in 2 study rabbits (20%) vs 10 controls (100%). In the second phase 37 rabbits were evaluable (8 died). Recurrent stricture was observed in 5 of the 18 study rabbits (27%) vs 14 of the 19 controls (73%). CONCLUSIONS: Halofuginone is effective in limiting the occurrence of de novo urethral stricture and recurrent stricture after visual internal urethrotomy. This antifibrotic molecule may become an important therapy to treat urethral stricture and/or recurrence following endoscopic manipulation of stricture in humans. PMID- 14532852 TI - Prospective analysis of 16S rDNA as a highly sensitive marker for bacterial presence in Peyronie's disease plaques. AB - PURPOSE: Previous studies have implicated an infectious agent induced pathogenesis in Peyronie's disease. To our knowledge an association with venereal diseases or other infectious diseases has not been demonstrated to date, although Peyronie's disease is an inflammatory disorder. In case of an infectious origin of the disorder bacteria or at least their fragments should occur in the plaque. We investigated prospectively the occurrence of 16S rDNA as a highly sensitive marker for the presence of bacteria in inflammatory processes. MATERIALS AND METHODS: In 19 patients with idiopathic Peyronie's disease biopsy of the tunica albuginea was sampled. A total of 16 men with no evidence of Peyronie's disease served as the control group. In these men tissue from the tunica albuginea was obtained during penile prosthesis implantation for erectile dysfunction or during a Nesbit procedure for congenital penile curvature. Screening for bacterial DNA was performed prospectively using a polymerase chain reaction targeting bacterial 16S rDNA. RESULTS: In the tunica albuginea specimen 16S rDNA was not detectable in patients with Peyronie's disease or in the control group. CONCLUSIONS: The results of this study do not indicate an association between Peyronie's disease and bacterial infection. PMID- 14532853 TI - Alterations of renal hemodynamics in unilateral ureteral obstruction mediated by activation of endothelin receptor subtypes. AB - PURPOSE: Unilateral ureteral obstruction (UUO) for 21 hours causes severe renal vasoconstriction. We examined the role of endothelin (ET)-A receptor in renal hemodynamic alterations induced by UUO. MATERIALS AND METHODS: Hemodynamic and clearance experiments were performed in 3 groups of anesthetized dogs. In group 1, 6 sham operated dogs received intrarenal infusion of the specific ET-A receptor antagonist BQ-610 (Peninsula Laboratories, Inc., Belmont, California), followed by infusion of the nitric oxide synthase substrate L-arginine. In the 7 group 2 dogs release of 21-hour UUO was followed by intrarenal infusion of BQ-610 and L-arginine. In the 5 group 3 dogs release of 21-hour UUO was followed by L arginine infusion. RESULTS: UUO caused marked decreases in renal blood flow (RBF) and glomerular filtration rate (GFR) in groups 2 and 3 compared with group 1. In group 1 BQ-610 and L-arginine infusion did not alter RBF or GFR. In contrast, BQ 610 infusion in group 2 after UUO release led to a significant increase in RBF and GFR as well as additional increases after L-arginine infusion. After UUO release in group 3 L-arginine infusion alone did not change RBF or GFR. CONCLUSIONS: After UUO release blockade of the ET-A receptor ameliorates renal vasoconstriction. The addition of L-arginine, which is a substrate for nitric oxide synthase, superimposed on ET-A receptor blockade confers a further decrease in renal vascular resistance, suggesting that the ET and L-arginine-nitric oxide systems are involved in renal hemodynamic alterations caused by UUO. PMID- 14532854 TI - Mitochondrial oxidative substrate selection in porcine bladder smooth muscle. AB - PURPOSE: Alterations in bladder smooth muscle (BSM) metabolism due to alterations in plasma lipid levels may be important with the increasingly high fat diets eaten by most Americans. To determine the susceptibility of BSM to lipotoxicity we examined the normal pattern of mitochondrial substrate selection in BSM and the ability of BSM to respond to changes in metabolic substrate provision. MATERIALS AND METHODS: BSM strips were incubated in 5 mM 1-13C-glucose and 0 to 5 mM 1,2-13C-acetate. The pattern of substrate use measured by 13C-nuclear magnetic resonance using BSM extracts. BSM was also cultured for 4 days to elicit changes in cell phenotype. RESULTS: At physiological levels of glucose and acetate about 50% of the substrate used by mitochondria was glucose. When acetate concentration was changed from physiological levels (0.1 mM) to pathophysiological levels (0.5 mM), BSM was able to increase the use of acetate, while sparing the use of glucose and intracellular substrates, likely lipids. Above 0.5 mM acetate BSM was unable to further use acetate. With increasing acetate use anaplerosis increased, consistent with a depletion of tricarboxylic acid cycle intermediates. After 4 days of organ culture BSM mitochondria used significantly more unlabeled intracellular substrates and less 13C labeled glucose than control bladder, consistent with metabolic adaptation to increase lipid use, such as what occurs with hyperlipidemia. CONCLUSIONS: We conclude that BSM has modest plasticity of the pattern of mitochondrial substrate selection and excess lipid provision may be able to induce lipotoxicity in BSM, resulting in impaired detrusor function. PMID- 14532855 TI - Alterations in voiding frequency and cystometry in the clomipramine induced model of endogenous depression and reversal with fluoxetine. AB - PURPOSE: Because serotonin (5-HT) in the central nervous system may inhibit bladder activity, we postulated that depression associated with altered 5-HT function might be associated with overactive bladder (OAB). We examined a rat model of endogenous depression caused by lowering 5-HT for effects on voiding frequency (VF) and awake cystometry (CMG), and examined the effect of reversal using the selective serotonin reuptake inhibitor fluoxetine. MATERIALS AND METHODS: Wistar rat pups were divided into 2 groups, namely clomipramine treated (CL) and saline control (SC). From postnatal days 8 to 21 each pup was injected with CL hydrochloride (22.5 mg/kg body weight) or an equal volume of saline. VF was assessed at 10 and 15 weeks. Behavioral correlates of depression were assessed using the forced swim test. At age 15 weeks CMG was performed. Fluoxetine (20 mg/kg daily) was administrated to a subset of SC/CL rats, and VF and CMG were repeated. RESULTS: In CL rats immobility was increased when rats were submitted to the forced swim test, indicating depression. CL rats voided more frequently than the SC group at 10 and 15 weeks but the difference was significant only in females. CMG of female CL group showed decreased bladder capacity, micturition volume and intermicturition contractions compared with the SC group. Treatment with fluoxetine reversed these changes, as in SC rats. CONCLUSIONS: These results support the hypothesis that OAB in a subgroup of depressed individuals may be associated with altered 5-HT function. It may explain reports of an association between depression and OAB. PMID- 14532856 TI - Collagen directly stimulates bladder smooth muscle cell growth in vitro: regulation by extracellular regulated mitogen activated protein kinase. AB - PURPOSE: Bladders clinically subjected to excessive pressure or distention demonstrate an altered extracellular matrix (ECM) composition. We determined how an altered collagen substratum might affect bladder smooth muscle cell (bSMC) growth in vitro and probed the mechanism of this response. MATERIALS AND METHODS: Primary culture rat bSMCs were seeded onto culture plates pre-coated with normal type I collagen (NC) or heat denatured type I collagen (DNC) under standard culture conditions. In separate experiments bSMCs from the 2 substrates were enzymatically released and changed to growth on normal collagen (NC-->NC or DNC- >NC) or denatured collagen (DNC-->DNC or NC-->DNC). At 24 hours proliferation was assessed by 3H-thymidine incorporation. Statistical significance in triplicate wells was determined by ANOVA. RESULTS: The proliferation of bSMCs on DNC was 5 fold greater than on NC (p <0.0001). Passage onto damaged collagen (DNC-->DNC) showed 2-fold further augmentation in proliferation (p <0.0001) but only a 50% decrease when NC was reintroduced (DNC-->NC) (p <0.001). Conversely replating on NC (NC-->NC) generated a 33% decrease in the already low proliferation rate (p <0.001) but 9-fold stimulation of proliferation when changed to damaged ECM (NC- >DNC) (p <0.0001). The mitogenic effect of damaged ECM on bSMC growth was abolished by specific inhibition of extracellular regulated kinase mitogen activated protein kinase signaling using PD98059. CONCLUSIONS: Damaged type I collagen (ECM) is mitogenic to bSMCs. The response is amplified by re-exposure to DNC. However, mitogenicity is only partially reversible by re-introducing NC. These results demonstrate striking bSMC responsiveness to ECM conformation. Signaling through the extracellular regulated kinase mitogen activated protein kinase pathway supports bSMC-ECM interaction. We speculate that remodeling the ECM in vivo may regulate bSMC growth. PMID- 14532857 TI - Calcineurin mediates bladder smooth muscle hypertrophy after bladder outlet obstruction. AB - PURPOSE: Bladder outlet obstruction (BOO) leads to compensatory bladder hypertrophy. However, the hypertrophy mechanism remains elusive. We report that calcineurin (Cn) is involved in bladder hypertrophy. MATERIALS AND METHODS: Partial BOO was surgically induced in 10-week-old female Wistar rats. The bladder was excised 2, 4 and 6 weeks following surgery in 9 rats each. Histological study was performed at each time point. Cn expression was examined by Western blot analysis. Myosin heavy chain expression was evaluated on gels stained with Coomassie blue. Primary cultured bladder smooth muscle cells were infected with recombinant adenoviruses encoding a constitutive active form of CnA (DeltaCnA), CnB and lacZ, and cell size was measured. RESULTS: In histological findings bladder smooth muscle hypertrophy was observed 2 and 4 weeks after surgery. However, the thickened muscles became thinner 6 weeks after BOO. CnA expression 2 weeks after BOO increased 3.2-fold compared with that of controls. Expression significantly decreased 4 and 6 weeks after surgery. In contrast, CnB expression was unchanged throughout hypertrophy development. Changes in myosin heavy chain expression correlated with changes in CnA. We observed significant hypertrophy in DeltaCnA and CnB over expressing smooth muscle cells. Moreover, FK506, which is a potent inhibitor of Cn, blocked hypertrophy in DeltaCnA and CnB over expressing smooth muscle cells. CONCLUSIONS: These data suggest that Cn has an important role in the induction of bladder smooth muscle hypertrophy. PMID- 14532858 TI - Effect of age on hydrogen peroxide mediated contraction damage in the male rat bladder. AB - PURPOSE: Recent studies introduced the concept that reactive oxygen species (ROS) may be a major factor in the progressive deterioration of bladder function induced by benign prostatic hyperplasia in men and following partial outlet obstruction in animals. We determined whether bladder contraction sensitivity to ROS changes with aging. Using H2O2 to simulate ROS damage we compared the sensitivity of the contractile responses of bladder smooth muscle isolated from young and elderly rats to H2O2. MATERIALS AND METHODS: Male 3-month-old (young) and 12-month-old (elderly) Sprague-Dawley rats were used for this study with 24 per group. Each rat was anesthetized and the bladder was excised. Two longitudinal strips were cut from the bladder body. Each strip was placed in individual 15 ml baths containing oxygenated Tyrode's solution at 37C. Each strip was stimulated at 32 Hz for 20 seconds with pulses 1 millisecond in duration at 80 V. After electrical field stimulation (EFS) the response to 20 microM carbachol and 120 mM KCl was determined. The bath solution was then exchanged for H2O2 solutions of 6 different concentrations including 0%, 0.0625%, 0.125%, 0.25%, 0.5% and 1.0%. After 1 hour of exposure to H2O2 the tissues were washed free of H2O2, and the response to EFS, carbachol and KCl were measured again. At the end of the experiment the strips were weighed and frozen at -70C for malondialdehyde analysis. RESULTS: The magnitude of the contractile responses of the young and elderly rats to all forms of stimulation were equal. Hydrogen peroxide caused a dose dependent decrease in the contractile responses of bladder strips to all forms of stimulation. Contractile responses to carbachol and KCl were more sensitive to H2O2 than to EFS. Contractile responses of bladder strips isolated from elderly rats were significantly more sensitive to H2O2 damage than strips isolated from young rats. Malondialdehyde generation of bladder strips isolated from elderly rats was significantly greater than those from young rats. CONCLUSIONS: It is suggested that aging increases the sensitivity of detrusor contraction to oxidative damage. PMID- 14532859 TI - Reperfusion injury of the rat bladder is worse than ischemia. AB - PURPOSE: Previous studies have demonstrated that in vivo and in vitro ischemia of the bladder results in decreased contractile responses. However, to our knowledge the effect of reperfusion following ischemia of the bladder is not known. MATERIALS AND METHODS: Adult male rats were subjected to bilateral bladder ischemia and varying periods of reperfusion. In vivo ischemia was created for 4 hours by reversibly clamping the 2 vesical arteries for 4 hours. Reperfusion was produced by removing the clamps and allowing the animals to recover for 1 day, 1 week or 1 month after surgery. Following recovery bladders strips were studied using field stimulation (FS), carbachol and KCl. The maximal contractile response and rate of response generated were recorded digitally and analyzed. RESULTS: The maximal responses to FS, carbachol and adenosine triphosphate (ATP) were not decreased by 4-hour ischemia alone, whereas the response to KCl was decreased significantly. The contractile responses to FS and KCl were significantly decreased after 1 day and 1 week of reperfusion. Responses after 1 month of reperfusion were increased significantly compared with responses after 1 week of reperfusion. The responses to ATP were not affected by ischemia or reperfusion. The contractile response to KCl was significantly more sensitive to ischemia than the responses to carbachol, ATP or FS, whereas the contractile response to FS was significantly more sensitive to reperfusion than the other forms of stimulation. CONCLUSIONS: This study demonstrates clearly that injury by reperfusion following ischemia is more detrimental than the effects of ischemia alone and FS contraction is the most sensitive form of stimulation to reperfusion damage. This study also demonstrates the ability of the bladder to recover from ischemic and reperfusion injuries. PMID- 14532860 TI - The N-terminal domain of vimentin alters bladder permeability. AB - PURPOSE: We assessed the effect of the vimentin amino terminal polypeptide (NT1) on barrier function of rabbit bladder epithelium. MATERIALS AND METHODS: The effect of NT1 on the properties of rabbit bladder epithelium were studied using Ussing chambers and electrophysiological methods. RESULTS: NT1 increased transepithelial conductance (Gt) in a voltage dependent manner. At a transepithelial voltage (Vt) of -70 mV (serosal solution ground) the addition of NT1 to mucosal solution did not result in a change in Gt. When Vt was clamped to 0 mV, there was a time dependent increase in Gt. The increase in Gt was reversed by clamping Vt back to -70 mV or by removing NT1 from the mucosal bath at 0 mV. The polypeptide acts primarily at the apical membrane with a conductance increase that is concentration dependent. Induced conductance is nonselective for small monovalent cations and anions. The ability of NT1 to increase membrane conductance was decreased in the presence of bath calcium. CONCLUSIONS: The data suggest that the amino terminus of vimentin can interact with the plasma membrane of bladder epithelium and increase ion permeability in a voltage dependent manner. PMID- 14532861 TI - Editorial: Membrane protein interactions in the bladder--charges of disorderly conduct. PMID- 14532862 TI - New way to deliver fluids: endoscopic jet injection into the beagle prostate. AB - PURPOSE: Local therapy of the prostate could have a role in the treatment of benign prostatic hyperplasia as well as prostate cancer. Endoscopic jet injection could simplify these treatments. We evaluated the tissue effects of endoscopic jet injection in an animal model. MATERIALS AND METHODS: An endoscopic jet injector was developed. In 12 beagle dogs the prostate was injected with single shot and continuous mode jet injection. Macroscopic and histological analysis was performed 0, 2 and 14 days after jet injection. RESULTS: Jet injection allowed complete penetration of the beagle prostate. The single shot technique resulted in only minimal tissue effects. However, the continuous mode led to major tissue damage, even complete aseptic necrosis, in the injected area. CONCLUSIONS: A new method for endoscopic application of fluids is described. Depending on the jet injection technique it could be used to apply pharmaceutical agents or plasmids, and/or induce aseptic necrosis. PMID- 14532863 TI - Effects of chronic administration of dehydroepiandrosterone on serum testosterone levels and prostatic tissue in rats. AB - PURPOSE: We evaluated the effects of chronic administration of dehydroepiandrosterone (DHEA) on serum total testosterone and DHEA sulfate (DHEAS), prostatic weight and histological features in the rat. MATERIALS AND METHODS: Wistar rats were divided into 2 groups. The first group of 9 rats received saline solution orally once daily during 5 days of the week for 10 months and the second group of 15 received 5 mg/kg daily DHEA in suspension orally in the same pattern. RESULTS: At the end of the 10-month study period total testosterone levels were higher in the DHEA group compared with controls (2.0 +/- 0.4 vs 0.8 +/- 0.2 mg/dl, p <0.0001) and DHEAS levels were also greater in the treated group compared with placebo (222.1 +/- 41.5 vs 2.0 +/- 0.3 mg/dl, p <0.0001). Weight of the prostate, testis and body did not differ between the groups (p >0.05). No differences between the 2 groups were noted in regard to the degree of hyperplasia, atrophy, a papillary component or the stromal-to-gland ratio (p >0.05). CONCLUSIONS: Oral ingestion of DHEA on a chronic basis in the rat increases serum DHEAS and total testosterone without any evident change in prostate weight or histology. PMID- 14532864 TI - Angiotensin II dependent testicular fibrosis and effects on spermatogenesis after vasectomy in the rat. AB - PURPOSE: Vasectomy induces testicular interstitial fibrosis in time dependent fashion and inhibits spermatogenesis. We investigated the contribution of angiotensin II (Ang II) to the development of interstitial fibrosis after vasectomy. MATERIALS AND METHODS: Bilateral vasectomy was performed in 8-week-old Wistar rats and the testes were harvested 1 to 24 weeks after vasectomy. Interstitial fibrosis was evaluated by Masson's trichrome staining. Western blotting and immunohistochemistry were done to examine the expressions of heat shock protein 47 (HSP47), HNE (4-hydroxy-2,3-nonenal) and transforming growth factor-beta1 (TGF-beta1). The antioxidative agent N-acetylcysteine (NAC), the Ang II type 1 receptor antagonist losartan or the Ang converting enzyme inhibitor enalapril was given orally for 24 weeks to vasectomized rats. Spermatogenesis was evaluated by testicular weight and the percent of haploid cells was analyzed by flow cytometry. RESULTS: Vasectomy significantly increased interstitial fibrosis (more than 8 weeks) and induced the expression of HSP47, HNE modified protein and TGF-beta1. TGF-beta1 and HSP47 immunoreactivity was localized to Leydig cells and fibroblasts. NAC or losartan but not enalapril inhibited the expression of these molecules induced by vasectomy. Increased interstitial fibrosis and impaired spermatogenesis were partially abrogated by NAC or losartan administration. CONCLUSIONS: There is Ang II type 1 receptor dependent fibrosis after vasectomy. Oxidative condition is considered to trigger and promote these fibrogenic processes. Ang II contributes to the regulation of intratesticular autocrine or paracrine functions after vasectomy. PMID- 14532865 TI - Effects of adenovirus mediated gene transfer to mouse testis in vivo on spermatogenesis and next generation. AB - PURPOSE: We directly injected DNA into mouse testes in vivo using an adenovirus vector to transfect testicular cells. We then analyzed the transfection efficiency, immunological problems and effects of gene transfer on spermatogenesis and the next generation. In this study we discuss the potential of gene therapy for male infertility. MATERIALS AND METHODS: A replication incompetent human adenovirus serotype 5 contained 2 deletions (E1 and E3 deletions) and was constructed such that the transgene was driven by the chicken beta-actin promoter to promote over expression of the downstream target gene (Lac Z). This adenovirus vector or control solution was injected into the interstitial space (intratesticular injection) or seminiferous tubules (intratubular injection) of the mouse testis. We investigated beta-galactosidase gene expression by X-gal (5-bromo-4-chloro-3-indolyl-beta-d-galactopyranoside) staining, the effects of gene transfer on spermatogenesis by evaluating the frequency of apoptotic cells by the TUNEL method, the inflammatory response on testes by detecting CD4 and CD8 positive cells immunohistochemically, and interleukin (IL)-6 and IL-8 by immunoblot analysis, epididymides sperm motility and the reproductive response of each mouse 3, 7, 14 and 28 days after injection. RESULTS: Intratesticular injection of adenovirus vector resulted in strong transgene expression in Leydig cells. In contrast, intratubular injection resulted in strong expression in Sertoli cells. Transgene expression was not detected in germ cells by either method. The peak of beta-galactosidase activity was on day 7, ie 0.674 +/- 0.20 (intratesticular) and 0.534 +/- 0.22 U (intratubular), and it decreased with time thereafter. The apoptosis index on day 7 was significantly higher in adenovirus injected groups than in noninjected groups, ie 0.46 +/- 0.20 vs 0.10 +/- 0.11 (intratesticular) and 0.78 +/- 0.31 vs 0.24 +/- 0.10 (intratubular). Transfected animals showed a slight mononuclear inflammatory response in the testes composed of CD4 and CD8 positive cells. Adenovirus vector stimulation resulted in the induction of IL-6 and IL-8 secretion in the testis. These immune responses subsided after day 7. There were no significant differences in the percent of motile sperm or the rate of abnormal sperm between the groups on any day after injection. Reproductive ability remained almost normal even after adenovirus mediated gene transfer with no effect observed in offspring. CONCLUSIONS: Our results suggest that although slight spermatogenic damage and inflammatory response caused by these methods may present problems, adenovirus mediated gene transfer may be effective for transfecting testicular somatic cells and applicable for in vivo gene therapy for male infertility in the future. PMID- 14532866 TI - Surveillance for elevated blood lead levels among children--United States, 1997 2001. AB - PROBLEM/CONDITION: Lead is neurotoxic and particularly harmful to the developing nervous systems of fetuses and young children. Extremely high blood lead levels (BLLs) (i.e.,> or =70 microg/dL) can cause severe neurologic problems (e.g., seizure, coma, and death). However, no threshold has been determined regarding lead's harmful effects on children's learning and behavior. In 1990, the U.S. Department of Health and Human Services established a national goal to eliminate BLLs >25 microg/dL by 2000; a new goal targets elimination of BLLs > or =10 microg/dL in children aged <6 years by 2010. REPORTING PERIOD: Information regarding children's BLLs comes from 1) National Health and Nutrition Examination Surveys (NHANES) conducted during 1976-1980, 1988-1991, 1991-1994, and 1999-2000; and 2) state child blood lead surveillance data for test results collected during 1997-2001. DESCRIPTION OF SYSTEM: CDC tracks children's BLLs in the United States by using both NHANES and state and local surveillance data. NHANES reports data regarding children aged 1-5 years; state and local surveillance systems report data regarding children aged <72 months. Because lead exposure in children varies among populations and communities, both surveys are needed to determine the burden of elevated BLLs among young children throughout the United States. NHANES uses highly standardized data-collection procedures and probability samples to gather information regarding the health and nutritional status of the civilian, noninstitutionalized U.S. population. State and local childhood lead surveillance systems are based on reports of blood lead tests from laboratories. State and local programs submit data to CDC annually. In this report, confirmed elevated BLLs are defined as one venous blood specimen > or =10 microg/dL or two capillary blood specimens > or =10 microg/dL drawn within 12 weeks of each other. RESULTS: The NHANES 1999-2000 survey estimated that 434,000 children (95% confidence interval = 189,000-846,000) or 2.2% of children aged 1-5 years had BLLs > or =10 microg/dL. For 2001, a total of 44 states, the District of Columbia (DC), and New York City (NYC) submitted child blood lead surveillance data to CDC. These jurisdictions represent 95% of the U.S. population of children aged <72 months and 97% of the nation's pre-1950 housing. The number of children tested and reported to CDC increased from 1,703,356 in 1997 (37 states, DC, and NYC reporting), to 2,422,298 in 2001 (44 states, DC, and NYC reporting). During that time, the number of children reported with confirmed elevated BLLs > or =10 microg/dL steadily decreased from 130,512 in 1997 to 74,887 in 2001. In 2000, the year targeted for national elimination of BLLs >25 microg/dL, a total of 8,723 children had BLLs > or =25 microg/dL. INTERPRETATION: Both national surveys and state surveillance data indicate children's BLLs continue to decline throughout the United States. However, thousands of children continue to be identified with elevated BLLs. The 2000 goal of eliminating BLLs >25 microg/dL was not met. Attaining the 2010 goal of eliminating BLLs > or =10 microg/dL will require intensified efforts to target areas at highest risk, evaluate preventive measures, and improve the quality of surveillance data. PUBLIC HEALTH ACTIONS: States will continue to use surveillance data to 1) promote legislation supporting lead poisoning prevention activities, 2) obtain funding, 3) identify risk groups, 4) target and evaluate prevention activities, and 5) monitor and describe progress toward elimination of BLLs > or =10 microg/dL. CDC will work with state and local programs to improve tracking systems and the collection, timeliness, and quality of surveillance data. PMID- 14532867 TI - Assisted reproductive technology surveillance--United States, 2000. AB - PROBLEM/CONDITION: In 1996, CDC initiated data collection regarding assisted reproductive technology (ART) procedures performed in the United States to determine medical center-specific pregnancy success rates, as mandated by the Fertility Clinic Success Rate and Certification Act (FCSRCA) (Public Law 102-493, October 24, 1992). ART includes fertility treatments in which both eggs and sperm are handled in the laboratory (i.e., in vitro fertilization and related procedures). Patients who undergo ART treatments are more likely to deliver multiple-birth infants than women who conceive naturally. Multiple births are associated with increased risk for mothers and infants (e.g., pregnancy complications, premature delivery, low-birthweight infants, and long-term disability among infants). REPORTING PERIOD COVERED: 2000. DESCRIPTION OF SYSTEM: CDC contracts with a professional society, the Society for Assisted Reproductive Technology (SART), to obtain data from fertility medical centers located in the United States. Since 1997, CDC has compiled data related to ART procedures. The Assisted Reproductive Technology Surveillance System was initiated by CDC in collaboration with the American Society for Reproductive Medicine, the Society for Assisted Reproductive Technology, and RESOLVE: The National Infertility Association. RESULTS: In 2000, a total of 25,228 live-birth deliveries and 35,025 infants resulting from 99,629 ART procedures were reported to CDC from 383 medical centers that performed ART in the United States and U.S. territories. Nationally, 75,516 (76%) of ART treatments were freshly fertilized embryos using the patient's eggs; 13,312 (13%) were thawed embryos using the patient's eggs; 7,919 (8%) were freshly fertilized embryos from donor eggs; and 2,882 (3%) were thawed embryos from donor eggs. The national live-birth delivery per transfer rate was 30.8%. The five states that reported the highest number of ART procedures were California (13,194), New York (11,239), Massachusetts (8,041), Illinois (7,323), and New Jersey (5,506). These five states also reported the highest number of live-birth deliveries and infants born as a result of ART. Overall, 47% of women undergoing ART-transfer procedures using freshly fertilized embryos from their own eggs were aged <35 years; 23% were aged 35-37 years; 19% were aged 38-40 years; 7% were aged 41-42 years; and 4% were aged >42 years. Among ART treatments in which freshly fertilized embryos from the patient's eggs were used, substantial variation in patient age, infertility diagnoses, history of past infertility treatment, and past births was observed. Nationally, live birth rates were highest for women aged <35 years (38%). The risk for a multiple birth delivery was highest for women who underwent ART-transfer procedures using freshly fertilized embryos from either donor eggs (40%) or from their own eggs (35%). Among women who underwent ART-transfer procedures using freshly fertilized embryos from their own eggs, further variation by patient age and number of embryos transferred was observed. Of the 35,025 infants born, 44% were twins, and 9% were triplet and higher order multiples, for a total multiple-infant birth rate of 53%. Patient's residing in states with the highest number of live-birth deliveries also reported the highest number of infants born in multiple-birth deliveries. INTERPRETATION: Whether an ART procedure was successful (defined as resulting in a pregnancy and live-birth delivery) varied according to different patient and treatment factors. Patient factors included the age of the woman undergoing ART, whether she had previously given birth, whether she had previously undergone ART, and the infertility diagnosis of both the female and male partners. Treatment factors included whether eggs were from the patient or a woman serving as an egg donor, whether the embryos were freshly fertilized or previously frozen and thawed, how long the embryos were kept in culture, how many embryos were transferred, and whether various specialized treatment procedures were used in conjunction with ART. ART poses a major risk for multiple births. This risk varied according to the patient's age, the type of ART procedure performed, and the number of embryos transferred. In addition, the increased risk for multiple births has a notable population impact in certain states. PUBLIC HEALTH ACTIONS: As use of ART and ART success rates continue to increase, ART related multiple births are an increasingly important public health problem nationally and in many states. The proportion of infants born through ART in 2000 that were multiple births (53%) was substantially higher than in the general U.S. population during the same period. Data in this report indicate a need to reduce multiple births associated with ART. Efforts should be made to limit the number of embryos transferred for patients undergoing ART. In addition, continued research and surveillance is key to understanding the effect of ART on maternal and child health. PMID- 14532868 TI - State-specific prevalence of selected chronic disease-related characteristics- Behavioral Risk Factor Surveillance System, 2001. AB - PROBLEM: High-risk behaviors and lack of preventive care are associated with higher rates of morbidity and mortality in the United States. Without continued monitoring of these factors, state health departments would have difficulty tracking and evaluating progress toward Healthy People 2010 and their own state objectives. Monitoring chronic disease-related behaviors is also key to developing targeted education and intervention programs at the national, state, and local levels to improve the health of the public. REPORTING PERIOD COVERED: Data collected in 2001 are compared with data from 1991 and 2000, and progress toward Healthy People 2010 targets is assessed. DESCRIPTION OF SYSTEM: The Behavioral Risk Factor Surveillance System (BRFSS) is an ongoing, state-based, telephone survey of persons aged > or =18 years. State health departments collect the data in collaboration with CDC. In 2001, participants in data collection included all 50 states, the District of Columbia, Guam, the U.S. Virgin Islands, and the Commonwealth of Puerto Rico. BRFSS data are used to track the prevalence of chronic disease-related characteristics and monitor progress toward national health objectives related to 1) decreasing high-risk behaviors, 2) increasing awareness of medical conditions, and 3) increasing use of preventive health services. For certain national objectives, BRFSS is the only source of data. RESULTS: BRFSS data indicate changes in certain high-risk behaviors from 1991 to 2001. Among the findings are substantial increases in the prevalence of obesity among adults aged > or =20 years. Among states, prevalence of persons classified as obese in 2001 ranged from 15.5% in Colorado to 27.1% in Mississippi. From 1991 to 2001, the median prevalence for all participating states and territories increased from 12.9% to 21.6%. In 1991, no state had an obesity prevalence of > or =20%; in 2001, 37 states had a prevalence of > or =20%. Percentage increases in prevalence of obesity, from 1991 to 2001, ranged from 24.9% in the District of Columbia to 140.2% in New Mexico. In 2001, substantial variations also existed among states and territories regarding prevalence of other high-risk behaviors and awareness of medical conditions. Ranges included, for no leisure-time physical activity, 16.5% (Utah) to 49.2% (Puerto Rico); cigarette smoking, 9.6% (Virgin Islands) to 31.2% (Guam); binge drinking, 6.8% (Tennessee) to 25.7% (Wisconsin); heavy drinking, 2.5% (Tennessee) to 8.7% (Wisconsin); persons ever told they had diabetes, 4% (Alaska) to 9.8% (Puerto Rico); persons ever told they had high blood pressure, 20% (New Mexico) to 32.5% (West Virginia); and persons ever told they had high blood cholesterol, 24.8% (New Mexico) to 37.7% (West Virginia). Substantial variations also existed among states regarding prevalence of using preventive health services. Ranges included, for persons aged > or =50 years ever screened for colorectal cancer by use of sigmoidoscopy or colonoscopy, 30.5% (Virgin Islands) to 62% (Minnesota); persons aged > or =65 years who received an influenza vaccination in the past year, 36.8% (Puerto Rico) to 79% (Hawaii); persons aged > or =65 years who ever received a pneumococcal vaccination, 24.1% (Puerto Rico) to 70.9% (Oregon). In 2001, 13 states, Guam, and the U.S. Virgin Islands used the women's health module. Ranges included, for women aged > or =18 years who had a Papanicolaou (Pap) smear test in the past 3 years, 79.8% (Virgin Islands) to 89.6% (Wisconsin); women aged > or =40 years who ever had a mammogram, 71.9% (Virgin Islands) to 93% (Rhode Island); and women aged > or =40 years who had a mammogram in the past 2 years, 57.2% (Virgin Islands) to 85.1% (Rhode Island). BRFSS data in 2001 also indicated variations by sex, race or ethnicity, and age group. Greater percentages of men than women reported cigarette smoking, binge drinking, heavy drinking, and were classified as overweight; greater percentages of women reported no leisure-time physical activity. Among racial or ethnic groups, greater percentages of black non Hispanics than other groups reported being told by a health professional they had high blood pressure and diabetes, and were classified as obese; greater percentages of white non-Hispanics than other groups reported being told they had high cholesterol. Among age groups, greater percentages of persons aged 18-24 years than those in older groups reported smoking cigarettes, binge drinking and heavy drinking; greater percentages of persons in older age groups than younger age groups reported being told they had diabetes, high blood pressure, and high blood cholesterol. Also, comparison of 2001 BRFSS data with 12 targets from Healthy People 2010 indicates that, in 2001, no state had met the targets for obesity, cigarette smoking, binge drinking, receiving a fecal occult blood test within the past 2 years, receiving annual influenza vaccinations, receiving pneumococcal vaccinations, and receiving Pap tests. Certain states had already met targets for no leisure-time activity, receiving a sigmoidoscopy or colonoscopy, having blood cholesterol checked within the past 5 years, and receiving a mammogram within the past 2 years. INTERPRETATION: BRFSS data in this report indicate that despite certain improvements, persons in a high proportion of U.S. states and territories continue to engage in high-risk behaviors and do not report making sufficient use of preventive health practices. Substantial variations (i.e., by state, sex, age group, and race/ethnicity) in prevalence of behaviors, awareness of medical conditions, and use of preventive services indicate a continued need to monitor these factors at state and local levels and assess progress toward reducing morbidity and mortality. PUBLIC HEALTH ACTIONS: BRFSS data can be used to guide public health actions at local, state, and national levels. For certain states, BRFSS is the only reliable source of chronic disease-related, risk-behavioral data. BRFSS data enable states to design, implement, evaluate, and monitor health-promotion strategies, targeting specific high-risk behaviors among populations experiencing high burdens of disease. BRFSS data continue to be key sources for assessing progress toward both national Healthy People 2010 objectives and state health objectives. PMID- 14532869 TI - Surveillance for health behaviors of American Indians and Alaska Natives. Findings from the Behavioral Risk Factor Surveillance System, 1997-2000. AB - PROBLEM/CONDITION: In the United States, disparities in risks for chronic disease (e.g., diabetes, cardiovascular disease, and cancer) and human immunodeficiency virus (HIV) and acquired immunodeficiency syndrome (AIDS) are evident among American Indians and Alaska Natives (AI/ANs) and other groups. This report summarizes findings from the 1997-2000 Behavioral Risk Factor Surveillance System (BRFSS) for health-status indicators, health-risk behaviors, and HIV testing and perceived risk for HIV infection among AI/ANs, compared with other racial/ethnic groups in five regions of the United States. REPORTING PERIOD COVERED: 1997-2000. DESCRIPTION OF SYSTEM: BRFSS is a state-based telephone survey of the civilian, noninstitutionalized, adult (i.e., persons aged > or =18 years) population. For this report, responses from the 36 states covered by the Indian Health Service administrative areas were analyzed. RESULTS: Region and sex-specific variations occurred in the prevalence of high-risk behaviors and health-status indicators. For example, the prevalence of current cigarette smoking ranged from 21.2% in the Southwest to 44.1% in the Northern Plains, and the awareness of diabetes was lower in Alaska than in other regions. Men were more likely than women to report binge drinking and drinking and driving. For the majority of health behaviors and status measures, AI/ANs were more likely than respondents of other racial/ethnic groups to be at increased risk. For example, AI/ANs were more likely than respondents of other racial/ethnic groups to report obesity (23.9% versus 18.7%) and no leisure-time physical activity (32.5% versus 27.5%). INTERPRETATION: The 1997-2000 BRFSS data demonstrate that health behaviors vary regionally among AI/ANs and by sex. The data also reveal disparities in health behaviors between AI/ANs and other racial/ethnic groups. The reasons for these differences by region and sex, and for the racial/ethnic disparities, are subjects for further study. However, such patterns should be monitored through continued surveillance, and the data should be used to guide prevention and research activities. For example, states with substantial AI/AN populations, and certain tribes, have successfully used BRFSS data to develop and monitor diabetes and tobacco prevention and control programs. PUBLIC HEALTH ACTIONS: Federal and state agencies, tribes, Indian health boards, and urban Indian health centers will continue to use BRFSS data to develop and guide public health programs and policies. The BRFSS data will also be used to monitor progress in eliminating racial and ethnic health disparities. Regional Indian health boards, tribal epidemiology centers, and Indian Health Service Area Offices can use the findings of this report to prioritize interventions to prevent specific health problems in their geographic areas. Moreover, tribes and other institutions that promote AI/AN health care can use the report to document health needs when applying for resources. PMID- 14532870 TI - Trichinellosis surveillance--United States, 1997-2001. AB - PROBLEM/CONDITION: Trichinellosis is a parasitic disease caused by tissue dwelling roundworms of the species Trichinella spiralis. The organism is acquired by eating Trichinella-infected meat products. The disease has variable clinical manifestations, ranging from asymptomatic to fatal. In the United States, trichinellosis has caused hundreds of preventable cases of illness and occasional deaths. The national trichinellosis surveillance system has documented a steady decline in the reported incidence of this disease, as well as a change in its epidemiology. REPORTING PERIOD COVERED: This report summarizes surveillance data for trichinellosis in the United States for 1997-2001. DESCRIPTION OF SYSTEM: Trichinellosis became a nationally reportable disease in 1966, but statistics have been kept on the disease since 1947. The national trichinellosis surveillance system is a passive system that relies on existing resources at the local, state, and federal levels. Cases are diagnosed based on clinical history with laboratory confirmation. Cases are reported weekly to CDC through the National Electronic Telecommunications System for Surveillance (NETSS). Detailed data regarding signs and symptoms, diagnostic tests, and food consumption are gathered by using a supplementary standardized surveillance form and are reported to CDC by fax or mail. This information is compared with NETSS data several times a year by CDC staff. Discrepancies are reviewed with the state health departments. The purpose of the surveillance system is to determine the incidence of trichinellosis, to maintain awareness of the disease, to monitor epidemiologic changes, to identify outbreaks, to guide prevention efforts, and to measure the effectiveness of those efforts. RESULTS: Although trichinellosis was associated historically with eating Trichinella-infected pork from domesticated sources, wild game meat was the most common source of infection during 1997-2001. During this 5-year period, 72 cases were reported to CDC. Of these, 31 (43%) cases were associated with eating wild game: 29 with bear meat, one with cougar meat, and one with wild boar meat. In comparison, only 12 (17%) cases were associated with eating commercial pork products, including four cases traced to a foreign source. Nine (13%) cases were associated with eating noncommercial pork from home-raised or direct-from-farm swine where U.S. commercial pork production industry standards and Regulations do not apply. INTERPRETATIONS: The majority of the decline in reported trichinellosis cases is a result of improved observance of standards and regulations in the U.S. commercial pork industry, which has altered animal husbandry practices resulting in reduced Trichinella prevalence among swine. PUBLIC HEALTH ACTIONS: Because of the change in epidemiology of trichinellosis and the continued occurrence of cases among consumers of wild game meat and noncommercial pork, more targeted public education is needed to further reduce the incidence of this disease. PMID- 14532871 TI - Training for infrainguinal bypass surgery. AB - BACKGROUND: Despite satisfactory results for surgery performed by trainees, vascular surgeons need to improve training methods to ensure that aspiring surgeons are adequately trained with less clinical exposure during fewer dedicated years of training. OBJECTIVES: To review the wide range of workshop, laboratory and seminar-room based methods available to train for the diverse range of skills required for distal arterial revascularisation. Training methods include anastomotic suturing skills with bench-top training apparatus, working with realistic plastic models and prosthetic conduits, cadaveric dissections and virtual-reality simulations. Many of these also provide excellent opportunities for objective assessment of technical skills and trainees' progress. DESIGN AND METHODS: A review of the literature on surgical education, surgical skills training and assessment. An evaluation of some of the apparatus, facilities, training curricula and courses, currently available to European trainees, is carried out. CONCLUSIONS: Many methods are now available to allow focused training for particular skills in non-clinical settings. Objective tools are also available that allow assessment of trainees at many levels or practicing surgeons. These technical skills assessment methods are important for trainees and surgeons who, in the future, will increasingly need to demonstrate competence in vascular surgery. PMID- 14532872 TI - Smoking cessation interventions in cardiovascular patients. AB - OBJECTIVES: To review current evidence for the effectiveness of smoking cessation interventions in cardiovascular patient populations. DESIGN AND MATERIALS: Studies were obtained from systematic reviews of the Cochrane Tobacco Addiction Review Group. Twelve Cochrane reviews related to secondary prevention in a variety of populations were used to extract eligible studies. Studies were eligible when they considered smokers diagnosed with symptomatic cardiovascular disease. METHODS: Data concerning the comparison between the intervention of interest and a placebo or standard treatment group were derived from eligible papers. Based on these data, Absolute Risk Reduction figures were calculated to express the effectiveness of each intervention on smoking cessation. RESULTS: We found 12 studies examining smoking cessation interventions in cardiovascular patients. Five studies reported significant results. No evidence was found for Nicotine Replacement Therapy or other pharmacology to be effective, neither for self-help materials, group, individual or telephone counseling. There is limited evidence for physician's advice and nurse-delivered interventions to be effective. CONCLUSIONS: Smoking cessation studies in cardiovascular patients are scarce. The available studies show very limited effects. The reasons for the lack of success of smoking cessation strategies in these patients remain unclear. Further research is needed to find effective cessation strategies for patients with cardiovascular diseases. PMID- 14532874 TI - Ruptured abdominal aortic aneurysm: endovascular repair is feasible in 40% of patients. AB - INTRODUCTION: Open repair of ruptured abdominal aortic aneurysm (rAAA) still has a high mortality. Endovascular aortic repair (EVAR) may be the way to improve survival rates. However, it is not clear how many patients with rAAA will be suitable for acute EVAR. METHODS: Between October 2000 and April 2002 all patients with acute symptomatic or ruptured AAA were assessed for EVAR on an intention-to-treat basis with emergency computed tomographic angiography (CTA). Patient and logistic characteristics were analysed. We used two commercially available aorto uni-iliac devices with a maximum proximal diameter of 28 and 34 mm. RESULTS: Five out of 26 patients were excluded for CTA and EVAR because of severe and persistent hypotension (3 pts) or logistic reasons (2 pts, both eligible). Twelve patients were found not eligible for EVAR due to unsuitable infrarenal aortic neck length (3 pts), neck diameter (1 pt) or a combination of both (8 pts). The remaining six patients were treated with EVAR. After 6 months no graft failure or aneurysm related deaths were recorded in the EVAR group. CONCLUSION: A total of 28% of patients with symptomatic or ruptured AAA was treated with EVAR. A potential 42% of patients could have been suitable for EVAR, if the correct devices had been in stock and all patients had been properly assessed. PMID- 14532875 TI - Rupture of infra-renal aortic aneurysm after endovascular repair: a series from EUROSTAR registry. AB - OBJECTIVES: Although small, the risk of rupture after EVAR remains a major concern. The aim of this study was to identify mechanisms of late aneurysm rupture after endovascular repair. METHODS: Patients who suffered a proven aneurysm rupture after EVAR were identified from the EUROSTAR (European Collaborators on Stent-graft Techniques for Abdominal Aortic Aneurysm Repair) registry. Complications preceding rupture were studied to identify common patterns and possible mechanisms of late rupture. RESULTS: A rupture was documented in 34 patients resulting in death of 21 (62%). Adverse events documented during previous follow-up in these patients included endoleak (30%), migration (18%), limb occlusion (12%) and kinking (12%). The findings at time of rupture were documented in 24 patients and including endoleak: Type III (10), Type I (9), Type II (1); stent-graft disintegration (2) and migration (3). Aneurysm diameter changes could be ascertained in 24 patients and had increased in only seven. CONCLUSION: The importance of graft-related endoleak, stent-graft disintegration and migration in the causation of aneurysm rupture was confirmed. Poor compliance with follow-up schedule was also identified as a common feature. However, the absence of complications in some patients, who attended regularly for follow-up, highlights the difficulty of predicting rupture after EVAR. PMID- 14532876 TI - The impact of endovascular stent grafting on reducing mortality rates after surgical treatment of abdominal aortic aneurysms. AB - OBJECTIVES: to investigate whether appropriate selection in patients with infrarenal abdominal aortic aneurysms (AAA) for transfemoral endovascular aneurysm management (TEAM) or open graft replacement (OGR) may decrease in hospital mortality rates (MR). DESIGN: analysis of a clinical series over three periods in an university vascular center. Conclusions of the second period were drawn and prospectively applied in a third period and compared. METHODS: during the period 1989-1994 only OGR was available (n=170). In the interval 1995-2000 either OGR or TEAM were carried out (n=454). During the period 01/2001-07/2002 the conclusions concerning selection of treatment modality were drawn and prospectively applied in 132 consecutive patients. MR were recorded and possible significant differences were checked. RESULTS: during the first period MR was 6.5%. Overall MR decreased to 3.7% in the second interval. Overall MR of the last period was improved to 1.5% (p<0.05). No patient died after OGR (0% vs 6.5%, p<0.04). As all patients with significant individual risk profiles were treated by TEAM, MR slightly increased (2.9%), but the difference remained insignificant (2.4% in period 2). CONCLUSIONS: risk adjusted selection of treatment modality influences the results after OGR significantly, thereby reducing overall MR of elective AAA treatment. PMID- 14532877 TI - Endotension as a result of pressure transmission through the graft following endovascular aneurysm repair--an in vitro study. AB - BACKGROUND: endovascular aneurysm repair (EVAR) significantly reduces, but does not abolish aneurysm sac pressure, possibly because of trans-fabric transmission. OBJECTIVE: to investigate how blood pressure is transmitted through different types of grafts into the aneurysm sac. DESIGN: experimental study, in vitro. METHODS: a latex aneurysm was inserted into an in vitro circulation model. The systemic mean pressure (SPmean) was varied from 50 to 120 mmHg. The grafts used for aneurysm exclusion were: thin wall polyethylene (PE), thick wall polyethylene (PE) and thin wall ePTFE. Mean aneurysm sac pressure (ASPmean) was measured, as was pulse pressure (ASPpulse). RESULTS: at an SPmean of 70 mmHg, the ASPmean was 34 +/- 0.8 mmHg (polyethylene knitted, thick wall), 30 +/- 1.0 mmHg (polyethylene woven, thin wall), and 17 +/- 0.6 mmHg (thin wall ePTFE). The ASPmean increased with SPmean, the relationship depending on the graft material. Stiffer grafts were associated with lower ASPmean and ASPpulse (p<0.001). CONCLUSIONS: the relationship between aneurysm sac mean pressure and systemic pressure (SP) depends on the graft material. These data highlights the need for further studies regarding endotension. PMID- 14532878 TI - Endovascular treatment of popliteal artery aneurysms. AB - OBJECTIVE: To present the results of the endovascular treatment of popliteal artery aneurysms. METHODS: From April 1999 to January 2002, 11 patients, aged 40 94 years, with 12 popliteal aneurysms were treated. Nine (75%) underwent an endoluminal repair, of whom three were done emergently due to an aneurysm rupture. Aneurysm diameter was 28-105 (mean 69) mm. A Hemobahn stent graft was inserted in six, Wallgraft in two and Passager in one case. RESULTS: During a mean follow-up of 14 (3-31) months, four (44%) thromboses occurred: two in the early postoperative period (30 days) and two during the late postoperative period. Two of the four occluded grafts were successfully reopened, and in the one a stenosis of the distal end of the stent graft was treated with balloon dilatation. Patency rates at 1 and 12 months were 64/47% (primary patency) and 88/75% (secondary patency), respectively. CONCLUSION: Initial experience with endovascular treatment of the popliteal aneurysm in high-risk patients yielded modest results. Larger number of patients and further follow-up time is necessary to evaluate the long-term results. PMID- 14532879 TI - Early and long-term results of conventional surgical treatment of secondary aorto enteric fistula. AB - OBJECTIVE: To retrospectively evaluate early and late results of surgical treatment of secondary aorto-enteric fistulas (SAEFs) with prosthetic excision and extra-anatomic bypass (conventional treatment) in a single centre institution (teaching hospital). MATERIALS AND METHODS: Between January 1990 and March 2002, 30 patients underwent conventional surgical treatment for SAEF. Data concerning these operations were collected in a dedicate database and 30-day mortality, patency and limb salvage rates were evaluated by mean of chi-square test and logistic regression analysis. Clinical and ultrasonographic follow-up was performed; late results were evaluated by mean of Kaplan-Meyer curves. RESULTS: Thirty day mortality rate was 26% (8 patients). Timing and sequence of interventions (simultaneous or staged, prosthetic excision or revascularization before) had no significative influence on perioperative mortality. There were six extranatomic bypass thromboses at 30 days, but no amputation. Mean duration of follow-up was 24 months; estimated 12- and 24-month survival rates were 60 and 50%, respectively. There were better results in terms of long-term survival in patients undergone prosthetic graft excision before. Primary patency rate was 62% and limb salvage rate was 95%, both at 24 months. Two prosthetic graft reinfections occurred during follow-up (9%). Cumulative reinterventions rate during follow-up was 18%. CONCLUSIONS: Conventional surgical treatment of SAEF permitted, in our experience, satisfactory early and long terms results, with fair rates of patency and limb salvage. Surgical timing and sequence do not seem to affect early results. PMID- 14532880 TI - Prevention of incisional hernia after aortic aneurysm repair. AB - BACKGROUND: Incisional hernia is a common late complication of elective abdominal aneurysm (AAA) repair. This paper describes a technique that could prevent the development of this condition. METHODS: Since Jan 2001, a polypropylene mesh has been sutured prophylactically in the pre-peritoneal space during abdominal closure after elective AAA repair. RESULTS: Twenty-eight consecutive elective procedures were performed. One patient died from a myocardial infarct 13 days after operation. Four patients (14%) had a wound infection (1 deep methicillin resistant Staphylococcus aureus (MRSA) infection and 3 superficial) that were treated successfully with antibiotics and dressings. One additional patient had a positive MRSA wound swab but required no treatment. Two patients required late re operations. One, who was on warfarin, required an urgent laparotomy for a leaking false aneurysm of the distal anastomoses 3 months after elective repair. A second patient had an anterior resection 18 months after aneurysm repair. Both re operations were uneventful. No patient has yet developed a clinically evident incisional hernia. CONCLUSIONS: These early data suggest that this mesh technique is a simple, safe and potentially effective method to decrease the incidence of incisional hernia following aortic aneurysm repair. PMID- 14532881 TI - Vascular endothelial cadherin expression in human carotid atherosclerotic plaque and its relationship with plaque morphology and clinical data. AB - OBJECTIVES: To determine the relationship between Vascular Endothelial (VE) cadherin expression in carotid plaques, carotid plaque morphology and clinical findings of carotid disease. MATERIALS AND METHODS: Fifty-three formalin-fixed, paraffin embedded specimens of human carotid atherosclerotic plaque obtained by endarterectomy and 20 normal postmortem arteries (control group) were studied. Thirty patients were symptomatic and 23 asymptomatic. The expression of VE cadherin was examined by an avidin-biotin immunoperoxidase technique using specific monoclonal antibodies against this molecule. We used a scale for the estimation of the expression of the VE-cadherin, in which negative expression was indicated by 0, weak expression by 1, and strong expression by 2. In serial sections we also determined the cellular phenotype of atherosclerotic plaques: i.e. the endothelial cells (F8), macrophage (CD68) and smooth muscle cells. Possible relations between variables in statistical analysis were examined by the chi-square test or Fisher's exact test. RESULTS: Expression of VE-cadherin was observed in small newly established vessels, particularly in areas with intense inflammatory infiltrations by macrophages and leucocytes. A strong expression of VE-cadherin was evident particularly in symptomatic instead in asymptomatic patients (43% vs. 13%, p=0.057), in high degree stenosis group (81% vs. 0%, p=0.005), and in patients with ischaemic infarct in brain scan (71% vs. 23%, p=0.021). On the other hand, there was no relation between molecule expression and plaque ultrasonic characteristics (echogenic or echolucent, p=0.499). Finally, there was a significant statistical correlation in the expression of VE cadherin and the histological type of the plaque, namely fibrotic and complicated plaques. Strong VE-cadherin expression was observed in 64% of complicated plaques instead of 6.5% in fibrotic plaques (p=0.001). CONCLUSION: An intense expression of VE-cadherin in carotid plaques is linked with plaque instability, high degree of stenosis and clinical events. This molecule seems to be a marker of progression of the atherosclerotic plaque. PMID- 14532882 TI - Carotid endarterectomy improves cognitive function in patients with exhausted cerebrovascular reserve. AB - OBJECTIVE: To investigate changes in cognitive function following carotid endarterectomy (CEA). DESIGN: Prospective study with controls. METHODS: CEA patients (n=159) were compared to a urology surgery control group (n=20). In CEA patients cerebrovascular reserve (CVR) was measured preoperatively. During surgery emboli and blood flow velocity in the middle cerebral artery were measured by transcranial Doppler (TCD) and cerebral oxygen saturation (CsO2) by near infrared spectroscopy. Cognitive function was measured preoperatively and at 5 days and 8 weeks postoperatively using a standardised computer battery of tests. RESULTS: Only 8% of patients had normal CVR bilaterally. The median number of emboli during CEA was 12 (range 0-181). On carotid clamping, TCD velocity fell a median of 41% and cerebral oxygen saturation by 5%. Attention deteriorated compared to controls 5 days following CEA (p=0.003) and this deterioration was related to the rise in TCD velocity on declamping (r=-0.3, p=0.002). Median attention reaction times improved significantly by 8 weeks (p=0.001) especially in patients' with severely impaired CVR before surgery (p=0.02). CONCLUSIONS: Attention improved at 2 months following CEA in patients with impaired CVR. CEA may offer more than reduced stroke risk to patients with impaired CVR. PMID- 14532883 TI - Patients' preferences for MR angiography and duplex US in the work-up of peripheral arterial disease. AB - PURPOSE: To measure patients' preferences for magnetic resonance (MR) angiography and duplex ultrasound (US) in the imaging work-up of peripheral arterial disease (PAD). METHODS: Ninety-eight patients, who underwent both MR angiography and duplex US, received a questionnaire by mail and their preferences for imaging work-up were assessed in a telephone interview. The questionnaire consisted of one question on which imaging test they preferred, a rating scale score ranging from 0 (not bothersome at all) to 10 (extremely bothersome), and specific questions on whether patients experienced discomfort due to the imaging work-up. Responses were presented as means and frequencies, and differences were tested by non-parametric tests. RESULTS: Half of the patient population (50%) had no preference for MR angiography or duplex US, 41% had a preference for MR angiography, and 9% had a preference for duplex US. The average rating scale scores for MR angiography (1.6) and duplex US (1.7) were not significantly different (p=0.53). Four out of 98 (4%) patients responded that they experienced pain during the MR angiography versus 18 (18%) for duplex US (p=0.001). Eight out of 98 (8%) patients responded that they were anxious during the MR angiography versus 1 (1%) for duplex US (p=0.02). CONCLUSION: The results suggest that the majority of patients have no preference between MR angiography and duplex US in the diagnostic work-up of PAD. Among patients who do have a preference, MR angiography was preferred over duplex US. PMID- 14532884 TI - Haemodynamic properties of PTFE femoropopliteal bypass grafts as determined by a new magnetic resonance technique. AB - OBJECTIVES: To apply the automatic three dimensional paraboloid (3DP) method for measurement of wall shear stress (WSS), blood flow, blood velocity, and cross sectional graft lumen area to magnetic resonance phase contrast velocity mapping data acquired from polytetrafluoroethylene (PTFE) suprageniculate 6 mm femoropopliteal bypass grafts to determine the biomechanical and haemodynamic properties immediately and three years after implantation. MATERIALS AND METHODS: Two patient cohorts, each of ten patients, with intermittent claudication were investigated by the 3DP method, one group (A) within 36 h after implantation of a femoropopliteal graft and the other group (B) 35 months after implantation. RESULTS: The mean blood flow was 5.9 ml/s (SD: 1.6), and WSS at peak systole was 2.2 N/m2 (SD: 0.6) 5 cm upstream to the distal anastomosis for group A, and 3.7 ml/s (SD: 1.9) and 1.9 N/m2 (SD: 0.4) for Group B. The WSS varied according to the angular position of the graft circumference. The implanted grafts were not circular but had a slightly elliptical circumference with a greater anterior/posterior diameter. CONCLUSION: The 3DP method is a precise tool with subpixel resolution for determining the biomechanical and haemodynamic properties of implanted PTFE grafts, and it can be used to assess graft function immediately after implantation. It is potentially applicable for routine graft surveillance. PMID- 14532885 TI - Rifampin and Triclosan but not silver is effective in preventing bacterial infection of vascular dacron graft material. AB - OBJECTIVES: To evaluate the efficacy of silver- or Triclosan-coated prosthetic material compared to Rifampin bonded Dacron concerning their resistance to infection following subcutaneous implantation and contamination with Staphylococcus aureus. DESIGN: Animal experimental study in mice. MATERIAL AND METHODS: Thirty-six C3H/HcN mice (Charles River Lab., Sulzfeld, Germany) with a weight between 24 and 27 g were randomised into six groups counting six animals each. Group I: control, gel-sealed dacron graft, group II: gel-sealed dacron graft and local contamination, group III: Intergard-Silver-prosthesis and contamination, group IV: silver/gel-sealed dacron prosthesis (test graft) and contamination, group V: Rifampin-bonded gel-sealed graft and contamination, group VI: Triclosan/collagen-coated dacron graft and contamination. Dacron graft material 0.8x1 cm was subcutaneously implanted in mice. Local contamination with 2x10(7)/0.2 ml S. aureus ATCC 25923 was carried out in groups II to VI. On day 14 the animals were killed and the grafts were explanted. The microscopic, histologic and microbiological evaluation of the graft material and the perigraft tissue was performed. RESULTS: In control group I no case of infection was detected. In group II, 6 of 6 animals showed infection. In group III (Intergard Silver) and group IV (silver/gel-test graft) were 6 of 6, in group V (Rifampin) only 1 of 6 grafts and in group VI (Triclosan) 4 of 6 grafts were infected. The difference between the low rate of infection in group V (Rifampin) in comparison to the completely infected groups III and IV (Silver) as well as the control group II was significant. Treatment of grafts with Triclosan could prevent infection only in 1/3 of the cases in group IV. CONCLUSION: Silver coating failed to prevent graft infection material. A potential antimicrobial property was evident for Triclosan whereas Rifampin-bonded grafts exhibit a significantly reduced infection rate. Thus, silver-coated vascular grafts cannot ensure protection from vascular graft infection. PMID- 14532886 TI - The incidence of hyperhomocysteinaemia in vascular patients. AB - INTRODUCTION: Hyperhomocysteinaemia has recently been identified as an important risk factor for atherosclerotic vascular disease. Screening for hyperhomocysteinaemia has been recommended, however, the incidence of hyperhomocysteinaemia in vascular patients is not known. AIMS: To determine the incidence of hyperhomocysteinaemia in vascular patients, to determine the relation of hyperhomocysteinaemia with folate, vitamin B12 levels and lipid profiles in vascular patients. To examine if there is a relationship between the degree of vascular injury and homocysteine concentration. METHODS: New vascular patients at The Queen Elizabeth Hospital were recruited and divided into peripheral, and aneurysmal presentations. Patients demographics were recorded, blood samples were taken for fasting lipid profile, and homocysteine concentration. Samples were also taken for vitamin B12, plasma and red cell folate levels. Sixty age and sex matched controls were included for comparison. RESULTS: One hundred and twenty-six patients have been recruited, (95 men and 31 women) with a median age of 68 years (61-74 years). The incidence of elevated homocysteine, and cholesterol levels was 33, 47 and 24%. The levels of vitamin B12 and folate were normal in all patients. Homocysteine was elevated in 27% of claudicants, 50% of patients with rest pain and 53% of patients with an aortic aneurysm. CONCLUSION: There is a high rate of hyperhomocysteinaemia in vascular patients with a higher incidence in patients with rest pain. There was also a high incidence of elevated homocysteine levels in patients with an abdominal aortic aneurysm. The rate of growth of these aneurysms is currently under review. Low folate or B12 concentrations is not the cause of raised homocysteine levels. PMID- 14532887 TI - Microvascular steal syndrome in the pedal bypass and free muscle transfer? PMID- 14532888 TI - A novel technique to control internal iliac artery back bleeding. PMID- 14532889 TI - Re: True juxta-anastomotic aneurysms in the residual infra-renal abdominal aorta. PMID- 14532897 TI - Genital and perianal warts: new treatment opportunities for human papillomavirus infection. AB - Human papillomaviruses are among the most common sexually transmitted diseases in the United States. Genital warts are a common phenotypic expression of human papillomaviruses, affecting 1% of the population; therefore, the obstetrician/gynecologist will invariably be required to advise and treat patients with this clinical manifestation. Issues essential in the diagnosis, counseling, and management of patients with genital warts will be examined, including epidemiology, transmission, molecular biology, and host immune response. This review will also provide the health care professional with a thorough examination of the new patient-applied treatment opportunities for anogenital condyloma, in addition to an overview of current provider-administered therapies, to assist in patient management. PMID- 14532898 TI - Molluscum contagiosum: the importance of early diagnosis and treatment. AB - Molluscum contagiosum is a viral infection that is becoming an increasing problem in sexually active individuals and in patients with human immunodeficiency virus. Although molluscum contagiosum lesions are generally self-limiting, it may take 6 months to 5 years for lesions to disappear. Furthermore, patients with weakened immune systems have increased difficulty in clearing lesions; therefore lesions typically persist for prolonged periods. Although there has been continued debate about whether molluscum contagiosum lesions should be treated or allowed to resolve spontaneously, many clinicians recommend treatment of genital molluscum contagiosum lesions to reduce the risk of sexual transmission, prevent autoinoculation, and increase patient quality of life. Treatment options for molluscum contagiosum include physician-administered and patient-administered therapies. Novel patient-administered treatment options allow administration in the privacy of a patient's home, providing added convenience and reducing patient embarrassment or stress. With the novel treatment opportunities currently available or in development, physicians are able to improve patient quality of life while providing patients with a convenient, well-tolerated, easily administered treatment regimen. This review summarizes the clinical diagnosis of molluscum contagiosum and provides a critical assessment of several current and emerging treatment options. PMID- 14532899 TI - Vulvar squamous cell carcinoma: guidelines for early diagnosis and treatment. AB - The incidence of vulvar squamous cell carcinoma (SCC), the most common vulvar cancer in women, is increasing worldwide. Furthermore, despite the increasing prevalence, this disease is commonly misdiagnosed by physicians. Unfortunately, late-stage vulvar SCC has a poorer prognosis compared with early vulvar neoplasia and requires aggressive therapy such as radical surgical intervention. Therefore, early diagnosis by physicians is critical to reduce both the mortality and morbidity rates of vulvar SCC. This review discusses the etiology, clinical characteristics, and diagnostic staging of vulvar SCC. Treatment options are reviewed, including novel topical immunotherapies such as imiquimod. PMID- 14532900 TI - New concepts in vulvodynia. AB - Vulvodynia is chronic vulvar burning/pain without clear medical findings. The etiology of vulvodynia is unknown and health care professionals should thoroughly rule out specific, treatable causes or factors such as dermatoses or group B Streptococcus infections. Vulvodynia is divided into 2 classes: vulvar vestibulitis syndrome is vestibule-restricted burning/pain and is elicited by touch; dysesthetic vulvodynia is burning/pain not limited to the vestibule and may occur without touch/pressure. After diagnosis, critical factors in successful patient management include education and psychological support/counseling. Unfortunately, clinical trials on potential vulvodynia therapies have been few. Standard therapy includes treating neuropathic pain (eg, tricyclic medications, gabapentin) thought to play a role. Additional therapies may be considered: pelvic floor rehabilitation combined with surface electromyography, interferon alfa, estrogen creams, and surgery. Importantly, any therapy should be accompanied by patient education and psychological support. Because definitive data on effective therapies are lacking, further clinical investigations of treatment options are warranted. PMID- 14532901 TI - Mitochondrial function: the heart of myocardial preservation. AB - Over the past several decades, it has become widely recognized that the mitochondria serve an important role in energy production and transfer to myocardial cells. More recently, mitochondria have been shown to play a key role in cell-death pathways by activating mitochondrial permeability transition pore formation and causing the release of several proteins, such as cytochrome c and proapoptotic factors. Although the molecular mechanisms for this process are still under investigation, important mitochondrial adaptations have been identified that can inhibit permeability transition activation and prevent apoptosis and necrosis. Specifically, myocardial preconditioning is an intriguing adaptation by which brief ischemia and reperfusion prime the mitochondria so that a subsequent prolonged period of ischemia is better tolerated. Mitochondrial ATP dependent potassium channels seem to play a critical role in triggering or mediating this protection. The objective of this review is to outline the role of the mitochondria in supporting myocardial energy under normal and ischemic conditions. It will also provide insight into the mechanisms by which mitochondrial signaling in preconditioning can protect the myocyte during subsequent prolonged ischemic periods. PMID- 14532902 TI - Xenogeneic cells and superantigen induce human T-cell activation in the absence of T-cell recognition of xenoantigen. AB - The extent of interaction between human T-lymphocytes and xenogeneic antigen presenting cells (APCs) is species-dependent. A successful interaction requires high-affinity receptor-ligand pairing across species and recognition of antigens presented by foreign major histocompatibility complex (MHC). A deficient human T lymphocyte response to xenogeneic cells is likely the result of a defect in these interactions. However, the requirements for a T-cell response to superantigen (SAg) may differ from those of other T-cell responses. Using irradiated murine splenocytes, which are believed to be incapable of eliciting human T-cell responses, and porcine aortic endothelial cells (PAECs) as the APC populations, we studied the human T-lymphocyte response to antigens presented by these cells. Direct proliferation of human T-lymphocytes to SAg presented by murine APCs was demonstrated; it was blocked by anti-human LFA-1 and anti-murine MHC class II but not by anti-human MHC class II. PAECs also presented SAg to human T-cells, generating a proliferative response greater than the primary response to porcine xenoantigen. Culture of human T-cells with murine splenocytes or PAECs and SAg Staphylococcus enterotoxin A (SEA) for 7 days primed human T-cells to proliferate in a secondary culture in response to autologous APCs. This autologous secondary response was human MHC class II-dependent and was inhibited by anti-human LFA-1, anti-human CD2, and anti-human CD98. Surprisingly, both of these responses were also blocked by anti-SEA, suggesting that despite vigorous washing, a small amount of functionally important SAg was carried over from primary to secondary culture, probably bound to the surface of T-cells. Xenogeneic APCs, even those that fail to stimulate human T-cells directly, can serve as APCs for primary human T-cell responses. After such interactions T-cells can develop secondary responses in autologous interactions based on retention of minute amounts of SAg. Such interactions may have important implications for xenotransplantation. PMID- 14532903 TI - In vitro and in vivo antipseudomonal activity, acute toxicity, and mode of action of a newly synthesized fluoroquinolonyl ampicillin derivative. AB - Compounds N-(6,7-difluoroquinolonyl)-ampicillin (AU-1) and N-(6-fluoroquinolonyl) ampicillin (FQ-1), synthesized by coupling of the carboxyl group of 6,7 difluoroquinolone (FP-3) and 6-fluoroquinolone (FP4), respectively, with the alpha-amino-group of ampicillin side chain, exhibit antipseudomonal activity similar to and lower acute toxicity than that of norfloxacin, whereas neither ampicillin nor the fluoroquinolone moieties, compound FP-3 or FP4, alone have such activity. Also, AU-1 and FQ-1 are active against tested clinical isolates of Pseudomonas aeruginosa that are highly resistant to norfloxacin, gentamicin, or both. The therapeutic efficacies of FQ-1 and norfloxacin were assessed and compared in neutropenic mice infected with a 90% lethal dose of P aeruginosa. Mice intraperitoneally administered FQ-1 (10 mg/kg) 4, 8, 24, and 48 hours after infection had survival rates as high as 80%, comparable to those of mice treated with norfloxacin at the same dosage and dosing schedule. The study of protoplast formation revealed that FQ-1 did not inhibit cell-wall biosynthesis but did induce cell filamentation of Bacillus subtilis at a level close to its minimal inhibition concentration. Both AU-1 and FQ-1 were able to intercalate into the double-stranded DNA. However, that FQ-1 lost such activity after it was treated with penicillinase suggests that the lactam-ring structure in ampicillin moiety of FQ-1 was hydrolyzed by penicillinase and that the hydrolyzed structure of FQ-1 does not own DNA-intercalation activity. PMID- 14532904 TI - Insulin and losartan reduce proteinuria and renal hypertrophy in the pregnant diabetic rat. AB - This study was designed to investigate the effect of hyperglycemia and angiotensin II (AngII) on renal hypertrophy and proteinuria in the pregnant diabetic rat. Secondary objectives were to evaluate changes in components of the renin-angiotensin axis and the effects of administration of losartan on pregnancy outcome. Fifty-three pregnant rats were allocated to 6 groups (1) nondiabetic controls (n = 12), (2) nondiabetic controls administered losartan (70-80 mg/kg/day; n = 10), (3) rats in which intravenous streptozotocin (STZ) was used to induce diabetes (55 mg/kg on day 10 of pregnancy; n = 10), (4) diabetic rats treated with losartan (n = 7), (5) diabetic rats treated with insulin (4 U/day; n = 7), and (6) diabetic rats treated with insulin and losartan (n = 7). Urinary protein excretion measured 4 days after STZ was 4 times greater in the rats with STZ-induced diabetes and significantly less in diabetic rats given losartan, insulin, or both. Postpartum kidney weight was greater in the rats with STZ induced diabetes (2.04 +/- 0.21 g) than in the controls (1.37 +/- 0.14 g; P <.05) and reduced in the diabetic rats given losartan, insulin, or both (1.57 +/- 0.22, 1.73 +/- 0.13, and 1.51 +/- 0.14 g, respectively; P <.05). Plasma levels of angiotensin II in rats given losartan were more than 3.5 times greater than those in controls (749 +/- 436, 596 +/- 323, 567 +/- 349, and 159 +/- 28 pg/mL; P <.001). Postpartum activity of angiotensin-converting enzyme was increased in the untreated diabetic rats compared with that in control rats (162 +/- 12 vs 117 +/- 16 nmol/mL/min; P <.05). This increase was abolished by treatment with losartan or insulin. The number of newborns and mean weight of each newborn was similar in all groups. In summary, administration of losartan or insulin prevented, in part, kidney hypertrophy and protein excretion in the diabetic pregnant rat. Losartan did not affect the number or weight of newborns. Because angiotensin II receptor blockers are contraindicated in pregnancy, good control of diabetes through the use of insulin should be advantageous. PMID- 14532905 TI - Activities of xanthine oxidoreductase and antioxidant enzymes in different tissues of diabetic rats. AB - Oxidative stress is an important pathogenic constituent in diabetic endothelial dysfunction. The aim of this study was to investigate whether an increase in oxidative stress related to xanthine oxidoreductase occurs in diabetes. Liver, brain, heart, and kidney xanthine oxidase (XO), xanthine dehydrogenase (XDH), antioxidant enzymes (glutathione peroxidase, superoxide dismutase, catalase), and nitrite levels were measured in control and early and late diabetic rat models. Although diabetes had no impact on liver XO and XDH activity, XDH activity in heart, kidney, and brain was significantly greater in late diabetic rats than in controls. Selenium glutathione peroxidase (GPx) activity was found to be lower in the liver, brain, kidney, and heart of late diabetic rats than in controls. The measured decrease in selenium GPx activity was also observed in early diabetic heart, kidney, and brain. No significant change was observed in liver, brain, and kidney copper/zinc superoxide dismutase (Cu/Zn SOD) activity in early and late diabetic rat models compared with that in controls, whereas heart Cu/Zn SOD activity was significantly decreased in both early and late diabetic rats. Liver and brain catalase activity remained similar among the different experimental groups, whereas increased heart and kidney catalase activity was observed in both early and late diabetic rats. Liver, kidney, and brain nitrite levels were found to be increased in early diabetic rat models compared with those in controls. These data suggest that the increased XDH and decreased selenium GPx activity observed in the later stages of diabetes leads to enhanced oxidative stress in the heart, kidney, and brain, resulting in secondary organ damage associated with the disease. PMID- 14532906 TI - Differential roles of hydrogen peroxide and hydroxyl radical in cisplatin-induced cell death in renal proximal tubular epithelial cells. AB - Reactive oxygen species (ROS) have been suggested as important mediators of cisplatin-induced acute renal failure in vivo. However, our previous studies have shown that cisplatin-induced cell death in vitro could not be prevented by scavengers of hydrogen peroxide and hydroxyl radical in rabbit renal cortical slices. This discrepancy may be attributed to differential roles of ROS in necrotic and apoptotic cell death. We therefore examined, in this study, the roles of ROS in necrosis and apoptosis induced by cisplatin in primary cultured rabbit proximal tubule. Cisplatin induced necrosis at high concentrations over a few hours and apoptosis at much lower concentrations over longer periods. Necrosis induced by high concentration of cisplatin was prevented by a cell permeable superoxide scavenger (tiron), hydrogen peroxide scavengers (catalase and pyruvate), and antioxidants (Trolox and deferoxamine), whereas hydroxyl radical scavengers (dimethythiourea and thiourea) did not affect the cisplatin induced necrosis. However, apoptosis induced by lower concentration of cisplatin was partially prevented by tiron and hydroxyl radical scavengers but not by hydrogen peroxide scavengers and antioxidants. Cisplatin-induced apoptosis was mediated by the signaling pathway that is associated with cytochrome c release from mitochondria and caspase-3 activation. These effects were prevented by tiron and dimethylthiourea but not by catalase. Dimethylthiourea produced a significant protection against cisplatin-induced acute renal failure, and the effect was associated with an inhibition of apoptosis. These results suggest that hydrogen peroxide is involved in the cisplatin-induced necrosis, whereas hydroxyl radical is responsible for the cisplatin-induced apoptosis. The protective effects of hydroxyl radical scavengers are associated with an inhibition of cytochrome c release and caspase activation. PMID- 14532907 TI - Experimental hypersensitivity pneumonitis: role of MCP-1. AB - Inhalation of Saccharopolyspora rectivirgula causes "farmer's lung" disease, a classic example of hypersensitivity pneumonitis (HP). Monocyte chemoattractant protein-1 (MCP-1) is increased in the bronchoalveolar lavage fluid of mice challenged with S rectivirgula, and S rectivirgula induces MCP-1 secretion by alveolar macrophages. We tested the hypothesis that MCP-1 and its receptor CC chemokine receptor-2 (CCR2) are essential to the development of experimental HP by treating mice with MCP-1 antibody and using CCR2(-/-) mice. Administration of anti-MCP-1 did not change the response to intratracheally administered S rectivirgula. CCR2(-/-) animals responded in a fashion similar to that of wild type animals to intratracheally administered.S rectivirgula. To determine the influence of the MCP-1-CCR2 interaction in vitro, we transferred S rectivirgula cultured spleen cells from S rectivirgula-sensitized mice, to naive recipients. Later, challenge of the recipients with intratracheal S rectivirgula and examination of both lung histology and bronchoalveolar lavage fluid characteristics were used to determine whether adoptive transfer had occurred. We found that cultured cells from CCR2(-/-) animals were fully capable of adoptive transfer. We conclude that interaction of MCP-1 with CCR2 is not necessary for the development of pulmonary inflammation in response to intratracheally administered S rectivirgula or cells able to adoptively transfer experimental HP. PMID- 14532908 TI - Coxsackievirus B1-induced chronic inflammatory myopathy: differences in induction of autoantibodies to muscle and nuclear antigens by cloned myopathic and amyopathic viruses. AB - Infection of susceptible strains of mice with the myopathic Tucson strain of coxsackievirus B1 (CVB1(T)) leads to the development of chronic inflammatory myopathy (CIM). The underlying mechanism of CIM appears to be immunopathic, but it is not known whether autoimmunity is involved. The objectives of this study were to determine whether autoantibodies are produced and whether they correlate with the pathology of CIM. Mice were infected with either a myopathic (MP1.23 or MP1.24) or an amyopathic (AMP2.17) CVB1(T) cloned virus. The two myopathic (MP) viruses cause CIM, whereas the amyopathic (AMP) virus, derived from a variant of the same parent, causes the same acute disease but does not cause CIM. Antimuscle IgG was found in 51% of MP1.23-infected and 58% of MP1.24-infected mice but in just 18% of mice infected with AMP2.17 and in 10% of controls (MP vs AMP: chi(2), P < or =.006). Several staining patterns were observed, indicating that autoantibodies of multiple specificities were produced. Antinuclear antibodies were found in 57% of MP1.23-infected and 27% of MP1.24-infected mice but were rare in mice infected with AMP2.17 (0%) or in controls (4%) (MP vs AMP: chi(2), P < or =.01). Antiviral-antibody titers were higher with MP virus than with AMP virus (ANOVA, P <.001). A trend toward an association between antiviral antibody or autoantibodies and the presence or severity of clinical measures of CIM was noted but was not significant. These data suggest that the autoantibodies do not mediate muscle disease but are an independent manifestation of an immunopathic response induced by infection with MP but not AMP CVB1(T). PMID- 14532909 TI - Left orbital frontal cortex volume and performance on the benton visual retention test in older depressives and controls. AB - Changes within the prefrontal cortex (PFC) have been associated with both mood disorders and with specific impairments in cognitive testing. The left PFC has been implicated in relational processing, that is, binding different pieces of information. We hypothesized that among older depressives and elderly controls, lower performance on one test of relational processing would be associated with smaller volume of the orbital frontal cortex (OFC). A total of 30 depressed and 40 control subjects were included in the study. All subjects were administered the Benton Visual Retention Test (BVRT). Subjects received a standardized magnetic resonance imaging, for which volumes of the OFC and total brain were calculated. We found that, controlling for age and education, total correct on BVRT was associated with left OFC volume normalized for total brain volume among the entire sample. For the depressed sample only, the number of perseverative errors was negatively associated with left OFC volume normalized for total brain volume after controlling for age and education. These results add to the literature linking mood and cognitive disturbances to the PFC. Future studies with a larger sample employing functional measures are warranted. PMID- 14532910 TI - Downregulation of the CCAAT-enhancer binding protein beta in deltaFosB transgenic mice and by electroconvulsive seizures. AB - Previous studies demonstrate that chronic, but not acute electroconvulsive seizures (ECS), increases levels of deltaFosB, a long-lasting transcription factor, in the hippocampus, and this effect correlates with the slow onset and long-lasting clinical effects of antidepressant treatment. To understand how deltaFosB mediates long-term plasticity in the hippocampus, we analyzed the gene expression profile of inducible transgenic mice expressing deltaFosB with a highly sensitive microarray assay and a customized computer analysis program. The CCAAT-enhancing binding protein-beta (C/EBPbeta) was identified as one of the genes downregulated by deltaFosB in the hippocampus. The downregulation of C/EBPbeta in the inducible deltaFosB transgenic mice was confirmed by other quantitative assays including real-time RT-PCR and low density dot blotting. Analysis of the C/EBPbeta expression in the hippocampus of rats treated with ECS revealed that the C/EBPbeta mRNA was also downregulated by chronic, but not acute ECS administration, the most effective treatment for depression. Given the reported role of C/EBPbeta in behavioral conditioning models, it is possible that the deltaFosB-mediated downregulation of C/EBPbeta in the hippocampus may be a molecular mechanism by which antidepressants alleviate some of the symptoms of depressed patients. PMID- 14532911 TI - Stress impairs alpha(1A) adrenoceptor-mediated noradrenergic facilitation of GABAergic transmission in the basolateral amygdala. AB - Intense or chronic stress can produce pathophysiological alterations in the systems involved in the stress response. The amygdala is a key component of the brain's neuronal network that processes and assigns emotional value to life's experiences, consolidates the memory of emotionally significant events, and organizes the behavioral response to these events. Clinical evidence indicates that certain stress-related affective disorders are associated with changes in the amygdala's excitability, implicating a possible dysfunction of the GABAergic system. An important modulator of the GABAergic synaptic transmission, and one that is also central to the stress response is norepinephrine (NE). In the present study, we examined the hypothesis that stress impairs the noradrenergic modulation of GABAergic transmission in the basolateral amygdala (BLA). In control rats, NE (10 microM) facilitated spontaneous, evoked, and miniature IPSCs in the presence of beta and alpha(2) adrenoceptor antagonists. The effects of NE were not blocked by alpha(1D) and alpha(1B) adrenoceptor antagonists, and were mimicked by the alpha(1A) agonist, A61603 (1 microM). In restrain/tail-shock stressed rats, NE or A61603 had no significant effects on GABAergic transmission. Thus, in the BLA, NE acting via presynaptic alpha(1A) adrenoceptors facilitates GABAergic inhibition, and this effect is severely impaired by stress. This is the first direct evidence of stress-induced impairment in the modulation of GABAergic synaptic transmission. The present findings provide an insight into possible mechanisms underlying the antiepileptogenic effects of NE in temporal lobe epilepsy, the hyperexcitability and hyper-responsiveness of the amygdala in certain stress-related affective disorders, and the stress-induced exacerbation of seizure activity in epileptic patients. PMID- 14532912 TI - Nociceptin/orphanin FQ increases anxiety-related behavior and circulating levels of corticosterone during neophobic tests of anxiety. AB - Intracranial administration of nociceptin/orphanin FQ (N/OFQ) increases circulating concentrations of adrenocorticotrophic hormone and corticosterone in unstressed rats, and elevates the responsiveness of these hormones during mild stress. Furthermore, N/OFQ and its cognate receptor are both abundant in a variety of limbic nuclei, and stress exposure decreases neuronal N/OFQ content in forebrain neurons. In light of these and other findings, we examined the potential involvement of N/OFQ in regulation of anxiety-related behaviors in rats. In the open field, elevated plus maze, and dark-light neophobic tests, intracerebroventricular N/OFQ (1.0 pmole-1.0 nmole) increased the expression of anxiety-related behaviors. Specifically, N/OFQ increased the latency to enter, decreased the number of entries into, and decreased the time spent in the exposed or brightly lit environments of all three tests. N/OFQ also enhanced thigmotactic responses in the open field test. The effects of diazepam and of the benzodiazepine inverse agonist FG 7142 were also assessed in independent groups of rats. In all three tests, the behavioral effects of N/OFQ resembled the anxiogenic actions of FG 7142, and contrasted with the anxiolytic actions of diazepam. N/OFQ administration also increased circulating concentrations of corticosterone during anxiety testing, in comparison with the concentrations in vehicle-treated controls. We conclude that N/OFQ administration is anxiogenic, and elevates responsiveness of the hypothalamic pituitary-adrenal axis during neophobic tests of anxiety. This supports the possibility that N/OFQ neurotransmission participates in processing of emotionally-salient and stressful stimuli, and suggests that normal functioning of the N/OFQ system may be important in physiological and psychological well-being. PMID- 14532913 TI - Attenuation of morphine tolerance, withdrawal-induced hyperalgesia, and associated spinal inflammatory immune responses by propentofylline in rats. AB - The activation of glial cells and enhanced proinflammatory cytokine expression at the spinal cord has been implicated in the development of morphine tolerance, and morphine withdrawal-induced hyperalgesia. The present study investigated the effect of propentofylline, a glial modulator, on the expression of analgesic tolerance and withdrawal-induced hyperalgesia in chronic morphine-treated rats. Chronic morphine administration through repeated subcutaneous injection induced glial activation and enhanced proinflammatory cytokine levels at the lumbar spinal cord. Moreover, glial activation and enhanced proinflammatory cytokine levels exhibited a temporal correlation with the expression of morphine tolerance and hyperalgesia. Consistently, propentofylline attenuated the development of hyperalgesia and the expression of spinal analgesic tolerance to morphine. The administration of propentofylline during the induction of morphine tolerance also attenuated glial activation and proinflammatory cytokines at the L5 lumbar spinal cord. These results further support the hypothesis that spinal glia and proinflammatory cytokines contribute to the mechanisms of morphine tolerance and associated abnormal pain sensitivity. PMID- 14532914 TI - Motor-learning impairment by amantadine in healthy volunteers. AB - NMDA receptor antagonists impair learning and memory in animal models, presumably by inhibiting long-term potentiation in the motor cortex. Human studies are limited and restricted by the paucity of safe NMDA antagonists. Here, we investigated the contribution of glutamatergic neurotransmission to the capacity of acquiring motor-adaptation learning in humans. In a double-blind design, 200 mg of amantadine (a low-affinity NMDA receptor channel blocker) or a matching placebo were given orally to groups of 14 and 13 human healthy young volunteers, respectively. Blood samples were collected 3 h after treatment to assay plasma concentrations, and the subjects were then tested using a motor-adaptation paradigm consisting of an eight-target-pointing task. To rule out drug-related generalized impairments such sedation, tests measuring motor dexterity and attention were also administered pre- and post-treatment. Comparison of the mean performance levels on the motor-adaptation task revealed that subjects in the amantadine group performed at a lower level than those in the placebo group, but this difference did not reach significance. Interestingly, however, despite plasma amantadine concentrations being relatively low, ranging from 2.09 to 4.74 microM (mean=3.3 microM), they nevertheless correlated negatively with motor learning. Furthermore, when the amantadine group was divided into low-performance and high-performance subgroups, subjects in the former subgroup displayed mean amantadine concentrations 36% higher than the latter subgroup, and performed significantly worser than the placebo group. No change in performance was found on the motor-dexterity and attention tests. Altogether, our results lend support to the hypothesis that normal NMDA receptor function is necessary for the acquisition of motor adaptation. PMID- 14532917 TI - [Prevalence of viral hepatitis type C in blood donors in Peru]. AB - In Peru there is limited information on the prevalence of HVC in the general population, for which reason we seek this information through blood donors in the different departments throughout the country. MATERIALS AND METHODS: Information was gathered from the Blood Bank Programs of the Ministry of Health establishments between 2000 and 2001, on which basis prevalences and their distribution by departments were obtained. RESULTS: The average prevalence, nationwide, was in the year 2000, 0.25% for HVC (0.08-0.48), 0.95% for HbsAg, 4.25% for Anti-HBc. In 2001, it was 0.60% for HVC (0.18-1.33), 0.9% for HbsAg and 4.51% for anti HBc. During 2000, for HVC, the result was 0.28% in the Coast and Jungle area 0.20% in the Sierra (Andean area) and during 2001 it was 0.89% in the jungle, 0.6% in the Coast and 0.46% in the Sierra. CONCLUSION: HVC prevalence in blood donors in Peru is low, however, it is necessary to extend the studies in the general population. PMID- 14532918 TI - [Omeprazole, amoxicillin and clarithromycin in the treatment of helicobacterpylori, in 7 and 10-day regimens]. AB - AIM: The most accepted treatment for infection by Helicobacter pylori is the proton pump inhibitor based therapy with two antibiotics. However, there is no consensus regarding the duration. The purpose here was to compare eradication percentages in the omeprazole+amoxicillin+clarithromycin regimen administered during 7 days versus 10 days and confront the results with a previous 14-day* experience in Peru. METHOD: Patients from the Central Military Hospital and Peruvian-Japanese Hospital evidencing chronic upper gastrointestinal tract symptoms were recruited. We excluded patients with peptic ulcer. Biopsies were taken for diagnosis, for urease and PCR tests, culture and coloring with silver. Omeprazole+clarithromycin+amoxicillin was used during 7 days versus 10 days. Control endoscopy was performed one month after treatment had been completed and molecular biology techniques were used to differentiate recurrences from new infections. Susceptibility to clarithromycin was assessed. RESULTS: 36 patients were included in each group. Eradication was the same in both groups: 86.1% (31/36). In several patients in whom the bacteria persisted, the same initial nucleus was found. In a previous study* using this same regimen during 14 days, a 93% eradication was obtained. 91.18% of our samples were susceptible to clarithromycin. CONCLUSIONS: In Peru, the omeprazole+clarithromycin+amoxicillin combination gives results higher than 80% in the eradication of infection by Helicobacter pylori. The 7 and 10 days regimens eradicated the bacteria in 86% of our patients. PMID- 14532919 TI - [Five-year survival prognosis factors in patients with resectable advanced serosa exposed gastric carcinoma]. AB - PURPOSE: To identify the main five-year survival prognosis factors in patients with resectable advanced serosa exposed gastric carcinoma, who have undergone radical gastric resection with limited and extended lymphadenectomy. MATERIAL AND METHODS: This retrospective study examined 137 patients with resectable advanced serosa exposed gastric carcinoma, in the Belen Hospital, Trujillo, Peru, between 1991 and 2000. RESULTS: The average age of the total series was 58.1 +/- 14.8 year (ranging from 20 to 84 years). This series was formed by 137 patients, of which 77 (56.2%) were male and 60 (43.8%) were female (M:F ratio=1.3:1). The univariate analysis using the log-rank test, showed that the following variables were significantly associated with five-year survival: lack of palpable mass (p=0.0308), serum haemoglobin concentration of 10 g/dl or higher (p=0.05) neoplasias located in the distal third of the stomach (p=0.0001) regional ganglionic condition N0-N1 (p=0.03), clinical stage II (p=0.0327) ganglionic dissection D2-D3 (p=0.0366) and curative intended surgery (p=0.000). The actual survival rate of the entire series after 5 years was of 16.3%. In the group subjected to lymphadenectomy D0-D1 (n=105) survival rate after 5 years was of 12.6%, while in the group subjected to D2-D3 (n=32) it was of 30.2% (p=0.0366). Five-year survival rate in patients who underwent curative intended surgery (n=56) was of 33.4% and those who had palliative surgery (n=81) had a five-year survival rate of 3.1% (p=0.000). CONCLUSIONS: Early detection, availability of curative surgery and the use of extended lymphadenectomy are factors that have an influence on the survival rate. These parameters should be considered for the staging of patients and their subsequent post-surgery assisting treatment. PMID- 14532920 TI - [New Beta-lactam agent in the treatment of intra-abdominal sepsis: double blind and randomized stage III study of ertapenem versus piperacillin/tazobactam]. AB - The clinical and safety efficacy of a new wide spectrum beta-lactam agent for most pathogen intra-abdominal infection germs is evaluated herein. Its chemical name is Ertapenem (MK-0826). Its pharmacokinetic characteristics and the known antibacterial spectrum enable the potential use of one daily dose in the treatment of infections by aerobic and anaerobic bacteria. This is a sub-group of patients that have been treated within a multinational, prospective, randomized, controlled and double-blind study, to compare the safety and efficacy of ertapenem (100% vs 88%) with piperacillin/tazobactam in patients that have undergone surgery due to complicated intra-abdominal infection, from April 1998 to October 1999, pursuant to the IDSA/FDA standards. Twenty local patients were evaluated from a total of 623 patients in 17 countries. Acute perforated appendicitis was the most frequent pathology in both groups. The recovery ratio was slightly higher in the group, which was administered ertapenem, with no documented clinical failure. This study shows the efficacy of ertapenem in the treatment of intra-abdominal infections using a single 1-gr/day dose, equivalent to 3.375 gr of piperacillin/tazobactam every six hours. Tolerance and safety were similar in both groups. No side effects, or mortality cases were registered. The results of this study indicate that ertapenem might be the therapeutic option to discard the combination of antibiotics or the use of multiple doses in intra abdominal infections. PMID- 14532921 TI - [Classification of stomach adenocarcinomas]. AB - Malignant stomach tumors include carcinomas, lymphomas, leimiosarcomas, carcinoids and other less frequent tumors. Adenocarcinoma has been classified in many different ways and by many different authors. Depending on its stage, early or advanced, on one side and according to the TNM staging system (Tumor, Nodes, Metastases) on the other. The early-stage adenocarcinoma, from the macroscopic point of view has been classified in I, IIa, IIc, IIb and III and combinations therefrom. Early-stage cancer has been denominated as O type and advanced cancer, which has been denominated by common practice, as Borrmann: I, II, III and IV, is now numbered using Arabic numbers 1, 2, 3 and 4. Type 5 is included, which would correspond to the non-classifiable carcinoma. Histologic classification according to Lauren, comprises intestinal, diffuse and the mixed or undifferentiated type which produces no mucus. According to Mulligan, it is classified as: pyloric glands and intestinal type cancer on one side and gastric type cell cancer on the other side. The WHO (World Health Organization) classifies them as: Papillary, tubular (tub.1, tub.2 and tub.3) signet ring cell, undifferentiated and mucinous adenocarcinoma. Nakamura, Kato and Hirota classify them as: differentiated and undifferentiated adenocarcinomas. Ming classifies them as: expanding type and infiltrating type. There is a tendency, when dealing with early-stage cancer, to group its forms in ulcerating carcinomas, vegetating carcinomas, localized gastritis-like and advanced-like carcinomas. The gastritis-like classification would correspond to form IIb of the initial classification of early-stage cancer. Broders' classification of Adenocarcinoma grade 1, 2, 3 and 4 is mentioned here as a classification solely on basis of the cellular differentiation. As historical classification, we include that of James Ewing. The above mentioned classifications relate to each other and are not excluding from the conceptual point of view. PMID- 14532922 TI - [Giant esophageal fibrovascular polyp]. AB - Fibrovascular polyps are extremely rare benign neoplasias of the esophagus, which usually originate in the lower cricoid area. They do not produce any discomfort in the patient for a long time, however it may make itself evident by the patient's regurgitation of the polyp, producing asphyxia or, more frequently, dysphagia. The case of a 58 year old male patient is presented herein, with a 9 month record of dysphagia, weight loss and intermittent melena. The barium x-ray showed a distended esophagus, with a tumor running from the upper esophageal sphincter to the cardia. The endoscopy confirmed the presence of a pediculated tumor, implanted in the cervical esophagus. Surgeons suspected the potential malignancy of the tumor and performed a transhiatal esophagectomy. The final pathologic diagnosis was giant fibrovascular esophageal polyp. PMID- 14532923 TI - [New case of human infection by a Pseudoterranova decipiens larva (Nematode, Anisakidae) in Peru]. AB - The purpose of this report is to divulge a new case of human anisakidosis in Peru, caused by a larva of the Pseudoterranova decipiens nematode found in a 17 year old female patient from Lima, who had ingested "cebiche" in a restaurant in the city of Ica, in the central coast of Peru. Approximately 4 hours after having ingested the food, the patient reported a feeling of uneasiness, with a nauseous sensation in the epigastric region, which intensified 5 hours later when she vomited. In the vomit content a live nematode was found, which was identified as larva L3 of P. decipiens. This is one of the etiologic agents of anisakidosis. PMID- 14532924 TI - [Gastric histoplasmosis simulating a malignant gastric ulcer]. AB - The case evaluated hereunder is that of a young patient, from a Histoplasmosis endemic area, reporting chronic abdominal pain. An upper gastric endoscopy evidenced the presence of gastric compromise, characterized by evident infiltrating ulcerous damage simulating a carcinoma. The final diagnosis was chronic disseminated histoplasmosis with gastric compromise. The patient presented as an underlying condition, a chronic alcoholic hepatopathy. Histoplasmosis must be included in the differential diagnosis of ulcerated and/or infiltrated lesions of the gastrointestinal tract in patients from endemic areas or susceptible to immunosuppression conditions. PMID- 14532925 TI - The appropriate use of sodium phosphates oral solutions. PMID- 14532926 TI - A randomized trial comparing seven-day ranitidine bismuth citrate and clarithromycin dual therapy to seven-day omeprazole, clarithromycin and amoxicillin triple therapy for the eradication of Helicobacter pylori. AB - OBJECTIVE: To assess Helicobacter pylori eradication after one week dual ranitidine bismuth citrate-clarithromycin (RBC-C) or triple omeprazole, clarithromycin and amoxicillin (OCA) therapy. METHODS: In this multicentre Canadian trial, H pylori-positive patients with functional dyspepsia or inactive peptic ulcer disease were randomized to open-label treatment with RBC-C (ranitidine bismuth citrate 400 mg plus clarithromycin 500 mg) or OCA (omezaprole 20 mg, clarithromycin 500 mg and amoxicillin 1000 mg), given twice a day for seven days. Treatment allocation was randomly assigned. H pylori infection was confirmed by positive 13C-urea breath test (13C-UBT). H pylori status was reassessed by UBT at least four and 12 weeks after treatment (negative: d13CO2 below 3.5 per mil). Intention-to-treat (ITT) eradication rates were determined for all patients with confirmed H pylori infection. Per protocol (PP) rate was determined for all patients treated with at least two evaluable follow-up visits. RESULTS: Three hundred five patients were included in the ITT and 222 in the PP analysis. The ITT eradication rates were 66% for RBC-C and 78% for OCA. The PP success rates were 84% for RBC-C and 96% for OCA. The difference for both ITT 12% (95% CI 2 to 22) and PP 12% (95% CI 4 to 19) were statistically significant, P=0.030 and P=0.007, respectively. Treatment was generally well tolerated. CONCLUSION: The eradication rate for the seven-day dual RBC-C regimen was lower than that for OCA. PMID- 14532927 TI - Electrocardiographic changes in elderly patients during endoscopic retrograde cholangiopancreatography. AB - BACKGROUND: Cardiorespiratory complications may occur during gastrointestinal endoscopy, and elderly people seem to be more vulnerable to these complications during endoscopic procedures involving the manipulation of abdominal viscera. OBJECTIVES: To determine the incidence of cardiac arrhythmias, changes in oxygen saturation, heart rate and blood pressure during endoscopic retrograde cholangiopancreatography (ERCP) via Holter monitoring in elderly patients older than 70 years of age. METHODS: Holter monitoring and 12-lead electrocardiograms were performed in 30 elderly patients undergoing ERCP and in 30 control subjects undergoing routine chest, abdomen, bone and upper gastrointestinal small bowel follow-through studies. A computerized nontriggered template system was used to analyze the electrocardiograms qualitatively and quantitatively. Arrhythmias, cardiac axis, conduction defects, pauses, ST segment changes, ectopic beats, oxygen desaturation and changes in blood pressure and rate-pressure product were evaluated. RESULTS: Increased heart rate, ST segment changes resulting from myocardial ischemia, oxygen desaturation and transient atrial and ventricular ectopic beats were frequent during ERCP compared with the control group. In one patient, transient left bundle branch block developed and this was attributed to pre-existing hypertension with cardiomegaly. One patient developed ventricular tachycardia and one other sinus bradycardia, but this was attributed to sick sinus syndrome. CONCLUSIONS: Transient myocardial ischemia and various cardiac arrhythmias are frequent in elderly patients undergoing ERCP. Appropriate noninvasive monitoring seems to be justified during this procedure. PMID- 14532928 TI - Cirrhotic cardiomyopathy: does it contribute to chronic fatigue and decreased health-related quality of life in cirrhosis? AB - Diminished health-related quality of life (HRQoL) and fatigue have been reported in patients with cirrhosis. The presence of cirrhotic cardiomyopathy and the attendant poor cardiac response to physical stress may affect HRQoL and contribute to fatigue in these patients. AIMS: To evaluate the contribution of cirrhotic cardiomyopathy to HRQoL and fatigue in ambulatory cirrhotic patients. METHODS: Thirty ambulatory cirrhotic patients (14 preascitic, Child-Pugh score 5.6 +/- 0.3; 16 ascitic, Child-Pugh score 9.1 +/- 0.5) underwent cardiopulmonary exercise testing and assessment of HRQoL, fatigue and depressive symptomatology. HRQoL and fatigue scores were correlated with liver disease severity, depressive symptomatology and parameters of cardiac function and exercise physiology. RESULTS: Subscales of all HRQoL measuring the effect of disease on the ability to perform physical, social and emotional roles were significantly impaired when compared with controls (P<0.001). The impact of fatigue, as assessed by the Fatigue Assessment Inventory, was greater in cirrhotics (4.46 versus 2.53 in healthy age-matched controls, P<0.01). Diminished HRQoL, impact of fatigue and vitality were related to depressive symptoms rather than to cardiac structural and functional abnormalities, which were present in cirrhotic patients. Poorer physical quality of life scores correlated with diminished mental health. CONCLUSIONS: Depression, rather than the presence of cirrhotic cardiomyopathy, may have contributed to the diminished HRQoL and vitality in patients with cirrhosis with or without ascites. PMID- 14532929 TI - Serous cystadenoma of the pancreas presenting as a third primary neoplasm. AB - Serous cystadenomas are the most common cystic neoplasms of the pancreas. They may occur solely or coexist with other neoplasms. A 10 cm mass involving the body of the pancreas was observed in the computed tomography of a 61-year-old man with a previous history of bladder and prostate carcinoma. Ultrasonography and computed tomography of the mass demonstrated multiple small cysts associated with a central calcified scar. A distal pancreatectomy was performed. Pathological examination confirmed the diagnosis of serous microcystic adenoma. This is the first report of a serous cystadenoma of the pancreas with two metachronous neoplasms. This feature should be kept in mind during the diagnosis and evaluation of patients with serous cystadenoma. PMID- 14532930 TI - Octreotide for enterocutaneous fistulas of Crohn's disease. AB - Crohn's disease is a chronic, life-long disease and has many troublesome complications. Fistulas are a part of Crohn's disease, and although there are many therapeutic modalities used in treating these fistulas they are only partially successful. As we are still very far from being able to cure it, the aim is to improve the patients' quality of life and to reduce the iatrogenic harm that may be caused by various treatment modalities. Somatostatin is an inhibitor of intestinal secretion and has been used to treat pancreatic and surgical fistulas. Therefore, we decided to treat enterocutaneous fistulas using somatostatin. METHODS: Five patients with Crohn's disease were treated with four daily injections of 300 microg octreotide. The total period of treatment was eight weeks. RESULTS: Closure of fistulas was achieved in four of the five patients. CONCLUSION: Somatostatin may have a role in treating Crohn's disease enterocutaneous fistulas and may prevent surgery or prolonged immunosuppressive therapy. PMID- 14532931 TI - Intestinal thromboangiitis obliterans in a woman: a case report and discussion of chronic ischemic changes. AB - Although traditionally regarded as a disease of distal extremities, mesenteric vasculature can also manifest thromboangiitis obliterans (TAO). There are 31 cases of intestinal TAO in the English literature and the majority of subjects are male. However, cases of women with TAO are becoming more common, coinciding with an increased incidence of smoking in this sex. We describe the sixth case of a female patient with classic extremity manifestations paralleled by paroxysms of abdominal angina. Intestinal TAO can mimic extremity disease of smoldering chronic ischemia punctuated by unpredictable acute episodes of gangrene. In the present case, chronic ischemia manifested as partial bowel obstruction due to stricture deformity of the ileum and profound adipocyte atrophy of mesentery. PMID- 14532934 TI - What's new on the net? Quality. PMID- 14532935 TI - Connecting the pieces of the cardiovascular care puzzle in Canada. PMID- 14532936 TI - The everlasting effect of primary percutaneous coronary intervention--even after transport? PMID- 14532937 TI - Hospitalization rates and length of stay for cardiovascular conditions in Canada, 1994 to 1999. AB - BACKGROUND: Cardiovascular diseases (CVDs) are a leading cause of hospitalization in Canada. An examination of recent trends in cardiovascular hospitalization rates across Canada is of considerable value and interest to health policy decision makers and administrators, clinicians and researchers. OBJECTIVES: To examine temporal trends and regional variation in hospitalization rates and length of stay for CVD conditions in Canada. METHODS: Hospital discharge data for fiscal years 1994/95 to 1999/2000 were used to identify all Canadians who were hospitalized with the most responsible diagnoses of acute myocardial infarction (AMI), congestive heart failure (CHF), angina and chest pain. Direct age- and sex standardized hospitalization rates were calculated by province and health region. Length of stay (LOS) for episodes of hospital care were adjusted for age, sex and cardiac procedures using ordinary least squares regression. RESULTS: Overall, AMI, angina and chest pain hospitalization rates increased 6%, 8% and 11%, respectively, between fiscal years 1994/95 and 1999/2000 and decreased by 7% for CHF in Canada. There was wide regional variation in cardiovascular hospitalization rates in Canada, with the greatest variation seen in CHF, chest pain and angina, and the least seen in AMI. There was a modest downward trend in adjusted LOS between fiscal years 1994/95 and 1999/2000. In general, patients hospitalized in provinces in western Canada and Ontario had shorter LOS for all conditions when compared with those in Quebec and the eastern provinces. CONCLUSIONS: AMI, angina and chest pain hospitalization rates in Canada increased between fiscal years 1994/95 and 1999/2000, while CHF rates declined. There is considerable regional variation in the cardiovascular hospitalization rates across the country that may be amenable to further interventional strategies. PMID- 14532938 TI - Interhospital transport for primary angioplasty improves the long-term outcome of acute myocardial infarction compared with immediate thrombolysis in the nearest hospital (one-year follow-up of the PRAGUE-1 study). AB - OBJECTIVE: Comparison of the long-term outcomes of three reperfusion strategies in patients with acute ST elevation myocardial infarction presenting to community hospitals. METHODS: One-year clinical outcomes were compared for 300 patients randomized in the PRimary Angioplasty in patients transferred from General community hospitals to specialized percutaneous coronary intervention Units with or without Emergency thrombolysis (PRAGUE-1) study to one of three treatment strategies: thrombolysis in a community hospital (group A, n=99); thrombolysis during immediate transportation for coronary angioplasty (group B, n=100); and immediate transportation for coronary angioplasty without thrombolysis (group C, n=101). RESULTS: Total mortality rates in group A, B and C patients were 18%, 12% and 13%, respectively (not significant). Nonfatal reinfarction occurred in 12%, 6% and 3% of patients, respectively (P<0.05). The combined endpoint (total mortality and nonfatal reinfarction rate) was reported in 30%, 18% and 16% of patients, respectively (P<0.05). In patients randomized within 2 h of the onset of symptoms, mortality rates were 18%, 3% and 8%, respectively (P<0.05). Additional revascularization procedures (percutaneous transluminal coronary angioplasty, coronary artery bypass graft surgery) were performed in 35%, 14% and 15% of patients, respectively (P<0.001). CONCLUSIONS: Primary angioplasty (even if delayed due to patient transportation to an interventional centre) is associated with better short- and long-term clinical outcomes than thrombolysis. The combination of the two strategies did not prove superior to coronary angioplasty alone. However, it may be superior in a subset of patients with early admission. The coronary angioplasty strategy decreases the need for revascularization procedures during the subsequent one-year follow-up. PMID- 14532939 TI - Infective endocarditis due to Streptococcus bovis in a series of nonaddict patients: clinical and morphological characteristics of 20 cases and review of the literature. AB - BACKGROUND AND OBJECTIVES: Controversy frequently exists about the actual frequency of Streptococcus bovis infective endocarditis (IE), its incidence of malignancy and its outcome. Consequently, the characteristics of S bovis IE were examined in an unselected population of nondrug-addicted patients. The literature was also reviewed. METHODS: Nondrug-addicted patients with S bovis IE were retrospectively reviewed. Clinically definite IE was diagnosed according to the Duke classification criteria at the single reference hospital for a defined population in northwestern Spain over a 13-year period. The clinical features, need for surgery and mortality rate of these patients were compared with those of other nondrug-addicted patients with IE examined during the same time period. RESULTS: Between 1987 and 1999, S bovis IE was diagnosed in 20 consecutive patients. This pathogen was responsible for 16.8% of the cases of definite IE in nondrug-addicted patients. Underlying conditions and embolic septic events were common. The aortic valve was the most common site of IE. Simultaneous involvement of two cardiac valves and moderate to severe regurgitation were more common in patients with S bovis IE. Colonic neoplasms were observed in 77% of patients. However, in-hospital mortality rate, need for in-hospital surgery and surgery during follow-up did not differ between patients with S bovis IE and the other nondrug-addicted patients with IE. CONCLUSIONS: In unselected patients, the rate of mortality due to S bovis IE is similar to that observed in IE due to other microorganisms. However, colonoscopic evaluation during admission and follow-up is required. PMID- 14532940 TI - Validation of a three-dimensional intravascular ultrasound imaging technique to assess atherosclerotic burden: potential for improved assessment of cardiac allograft coronary artery disease. AB - BACKGROUND: Serial analysis of intracoronary ultrasound images is limited by difficulty with spatial registration and inability to assess the full extent of vascular disease. Three-dimensional (3D) imaging of coronary arteries can potentially overcome these limitations. OBJECTIVES: To assess the feasibility of using a PC-based 3D rendering technique to assess atherosclerotic burden. METHODS: To define the accuracy of 3D intravascular ultrasound (IVUS) measurements in vitro, six porcine iliac arteries and nine human cadaveric iliac arteries were pressure fixed and imaged with a commercial IVUS system. 3D datasets of the arteries were constructed, and measurements were correlated with histomorphometry. In vivo studies of 53 arterial segments (19 right coronary, 26 anterior descending and eight circumflex) were scanned in 18 patients, one month to nine years post-transplantation and correlated to corresponding angiographic images for the presence of atherosclerosis. RESULTS: Porcine artery length and volume measurements by IVUS showed a high degree of correlation with histomorphometry measurements (r=0.99, P<0.0003 and r=0.99, P<0.0001, respectively). Human arterial length, total artery volume and lumen volume measurements were similarly correlated (r=0.99, P<0.0001, r=0.99, P<0.0001 and r=0.98, P<0.0001, respectively). For plaque volume, r=0.84, P<0.05. In vivo 3D IVUS scans demonstrated atherosclerotic lesions in nine of 18 patients, compared with five detected by angiography alone. CONCLUSIONS: 3D IVUS imaging allows rapid and accurate measurement of arterial length, volume and plaque dimensions in addition to lumenal area and can demonstrate the full extent of atherosclerotic pathology. Because of its superior reproducibility, this technique may be used to assess the progression of coronary artery disease and allow for more accurate evaluation of interventions aimed at preventing or retarding coronary artery disease. PMID- 14532941 TI - Incidence, coronary risk profile and angiographic characteristics of prediabetic and diabetic patients in a population with ischemic heart disease. AB - BACKGROUND: Recently, the threshold of fasting blood glucose indicating diabetes mellitus was lowered to 7.00 mmol/L (126 mg/dL) and the term 'impaired fasting glucose' (IFG; fasting blood glucose ranging from 6.11 mmol/L to 6.99 mmol/L or from 110 mg/dL to 126 mg/dL) was introduced to define a prediabetic state. OBJECTIVE: To evaluate the incidence of the above states in a Canadian population with suspected coronary artery disease and to compare their risk profiles and angiographic status to normoglycemic subjects. PATIENTS AND METHODS: Revision of the database of 1108 consecutive patients (793 males and 315 females; mean age 58.1+/-9.8 years) undergoing clinical, biochemical and elective angiographic studies because of suspected coronary artery disease. RESULTS: One third of the patients had either IFG (8.5%), or were diabetics (24.2%). Unlike the 747 normoglycemic patients, both IFG (n=94) and diabetic (n=267) subjects showed an insulin resistance profile, with abdominal obesity, and dislipidemia characterized by high triglycerides in the presence of low high density lipoprotein-cholesterol and high normal or elevated blood pressure. Both prediabetics and diabetics had a significantly higher homeostatic model assessment insulin resistance index than normoglycemics (P<0.0001), the index also being higher for diabetics than for prediabetics (P<0.0001). Coronary atherosclerosis was documented in most patients of the three groups and was significantly more severe in diabetics than in IFG patients (P=0.0359) or normoglycemics (P=0.0069), with no differences between the former two groups. CONCLUSIONS: As expected, the new definitions identify more patients with impaired homeostasis than earlier criteria. IFG patients have similar coronary risk profile as diabetics, suggesting the need for similar coronary precautions. PMID- 14532942 TI - Milk whey protein decreases oxygen free radical production in a murine model of chronic iron-overload cardiomyopathy. AB - BACKGROUND: Chronic iron overload is a major cause of organ failure worldwide, but its pathogenesis remains to be elucidated. OBJECTIVES: To examine in an experimental murine model of iron-overload cardiomyopathy the relation between milk whey protein and, first, the production of reactive oxygen free radical species and, second, antioxidant reserve status. METHODS: B6D2F1 mice were randomly assigned to four treatment groups (n=8 per treatment group): placebo control; iron only; whey only; and iron with whey. Reactive oxygen free radical species in the heart were quantified by the cytotoxic aldehydes malondialdehyde (MDA), 4-hydroxy-nonenal (HNE) and hexanal, while antioxidant reserve status was quantified by glutathione (GSH) and glutathione peroxidase (GPx) activity in the heart tissue. RESULTS: Significantly decreased concentrations (pmol/100 mg wet weight tissue) of MDA (2468+/-261), HNE (912+/-38) and hexanal (5385+/-927) were observed in the heart tissue of the group receiving iron with whey, in comparison with the iron-only treatment group (MDA 9307+/-387, HNE 1416+/-157, hexanal 14,874+/-2955; P<0.001). Significantly increased GPx (141+/-38 IU/L) and GSH (521+/-136 IU/L) activity were observed in mice receiving iron with whey, in comparison with mice receiving iron only (GPx 100+/-10 IU/L, GSH 446+/-33 IU/L; P<0.001). CONCLUSION: Mice receiving iron treatments with whey supplementation had significantly lower concentrations of cytotoxic aldehydes and significantly higher cardiac levels of GPx and GSH activity than did iron-only treated mice. Additional basic research is warranted to examine the exact mechanisms by which milk whey protein protects the heart. PMID- 14532943 TI - National Cholesterol Education Program Adult Treatment Panel III guidelines and obesity: implications for Canada. AB - BACKGROUND: The National Cholesterol Education Program Adult Treatment Panel (ATP) III recommendations incorporate new evidence for treating elevated low density lipoprotein cholesterol. OBJECTIVES: Comparisons between the prevalence of drug-eligible Canadians under the old ATP II and the new ATP III guidelines were made, and the impact of obesity on current and future drug eligibility was explored using various models. METHODS: Participants from the Canadian Heart Health Surveys (1986 to 1992; n=17,728; 20 to 74 years of age) were assigned to therapeutic lifestyle change or drug-eligible groups in the event of elevated low density lipoprotein cholesterol. Body mass index was used to classify participants as having normal weight, or as being overweight or obese. The prevalence of overweight and obese status for 2001, 2006 and 2011 were projected from past trends by linear regression. Population attributable risk was used to model reductions or increases in the prevalence of obesity in drug-eligible participants using several nationally representative population health surveys. RESULTS: In 2001, an additional 1.1 million Canadians were drug-eligible under ATP III (16.0% of men, 9.5% of women), compared with ATP II (7.7% of men, 7.7% of women). Drug eligibility was elevated in overweight participants (men: OR=1.87 [1.51 to 2.31]; women: OR=1.60 [1.13 to 2.28]) and the obese (men: OR=2.86 [1.86 to 4.38]; women: OR=2.28 [1.63 to 3.18]) versus normal weight participants. The population attributable risk was higher in men (overweight 22.6%, obese 11.5%) than in women (overweight 9.4%, obese 9.2%). IMPLICATIONS: A 10% reduction in overweight and obesity prevalence could have prevented 69,530 cases of drug eligibility in 2001. On the other hand, by 2011 over one million Canadians will be drug-eligible because of an elevated body mass index, if the recent trends in overweight and obese status continue. PMID- 14532944 TI - An unusual case of a giant aneurysm of the ascending aorta. PMID- 14532945 TI - Churg-Strauss syndrome with myocarditis manifesting as acute myocardial infarction with cardiogenic shock: case report and review of the literature. AB - A patient with a two-year history of worsening asthma presented with chest pain and shortness of breath. She developed cardiogenic shock. Analysis of blood chemistry detected increased troponin I concentration. Her electrocardiographic changes were consistent with a diagnosis of anteroseptal myocardial infarction. However, angiography showed normal coronary arteries. Left ventriculography showed severe mitral regurgitation and global hypokinesis. Peripheral eosinophilia was detected. Subsequent endomyocardial biopsy showed myocarditis with prominent eosinophil and plasma cell components. Churg-Strauss syndrome was diagnosed based on her history of asthma, evidence of peripheral eosinophilia and results of endomycardial biopsy. Treatment with a high dose of corticosteroids was initiated. As symptoms of heart failure improved - without recurrence of cardiac and respiratory symptoms - the dose of corticosteroids was gradually reduced. Eight months after her original presentation, she developed urticarial lesions on her abdomen and legs, with muscle soreness but no other associated symptoms. She was treated with a combination of prednisone and dapsone. After the diagnosis of Churg-Strauss syndrome, the patient remained symptom free with a normal ejection fraction for 15 months while taking prednisone. PMID- 14532946 TI - Subaortic stenosis: recurrence of a fibrous ring after 28 years. AB - Aortic stenosis is a common cardiac problem. Morphological aortic stenosis can be due to valvular, subvalvular and supravalvular causes. Subvalvular causes include subaortic rings and membranes, which usually manifest at a young age, depending on the size of the ring and the degree of obstruction. Recurrent (post-operative) stenosis is a rare potential problem. A case of recurrent subaortic stenosis due to a subaortic ring, 28 years after the initial surgical excision of the ring is presented. PMID- 14532947 TI - The economics of medical education. PMID- 14532948 TI - Why JOGC has "The Pharmacotherapy File". PMID- 14532949 TI - Metastatic gastrointestinal tract cancer presenting as ovarian carcinoma. AB - OBJECTIVES: (1) To assess if there has been an increase of metastatic gastrointestinal (GI) cancers presenting as ovarian cancer; (2) to determine from which areas of the GI tract malignancies are most frequently associated with ovarian metastasis; (3) to determine if a higher proportion of premenopausal women present with an ovarian tumour of GI tract origin. METHODS: A retrospective chart review of women presenting to the Hotel-Dieu de Quebec Hospital from January 1993 to June 2002, searching for the diagnosis of ovarian metastasis of GI origin at the time of primary surgery. RESULTS: Colon cancer accounted for 65% of the malignancies found to have ovarian metastases at the time of primary surgery, with an increased percentage of cases in recent years. Metastatic GI cancer mimicking ovarian primary was equally as frequent in pre- and postmenopausal women. The use of the cancer antigen (CA) 125/carcinoembryologic antigen (CEA) ratio may help to discriminate GI carcinoma from ovarian carcinoma. CONCLUSION: Because GI tumours may present as advanced ovarian cancer, increased vigilance is required in order to make the right diagnosis and offer the best treatment. PMID- 14532950 TI - Maternal serum screening: practice patterns of physicians in Newfoundland. AB - OBJECTIVE: To compare the utilization of the second trimester maternal serum screen (MSS) of a-fetoprotein, human chorionic gonadotrophin, and unconjugated estriol, in Newfoundland, by practice location, training, and gender. METHODS: Four hundred eighteen anonymous self-reported questionnaires were mailed out to all practising family physicians, general practitioners, and obstetricians in Newfoundland, who were identified through the provincial medical board. The survey included questions on demographic characteristics, provision of antenatal care, gestational age at which MSS is ordered, reasons for offering or not offering MSS, and the use of routine antenatal ultrasound. Categorical data were analyzed using chi-square and Fisher exact tests, as appropriate. RESULTS: Overall, 63% of physicians responded to the survey. Forty percent of respondents had an urban practice. Female physicians, regardless of specialty, were more likely to offer MSS to their patients (89% vs. 78%; P = 0.04), whereas family physicians and obstetricians were more likely to offer screening than general practitioners (85% vs. 83% vs. 25%; P = 0.02). Among physicians offering MSS, 54% offered it only to women 35 years and older. Practice location did not affect whether a woman was offered MSS (P = 0.41). Twenty-five percent of family physicians offering MSS did not offer it at the appropriate gestational age of 15 to 20 weeks. Ninety-four percent of pregnant women were routinely offered an ultrasound during pregnancy. CONCLUSION: The utilization of MSS in Newfoundland is affected by physician training and gender, but not by practice location. Further education of physicians is required to ensure appropriate use and timing of this screening test. PMID- 14532951 TI - Diclectin therapy for nausea and vomiting of pregnancy: effects of optimal dosing. AB - OBJECTIVES: (1) To quantify rates of suboptimal use of pyridoxine hydrochloride doxylamine (Diclectin); and (2) to study responses to optimal doses of Diclectin in women previously taking a suboptimal dose. METHODS: Women who called the Motherisk NVP helpline, and were taking only Diclectin (vitamin B6 10 mg and doxylamine 10 mg), were enrolled in the study and assessed for the severity of nausea and vomiting of pregnancy (NVP) with the Motherisk-PUQE (pregnancy-unique quantification of emesis and nausea) scoring system. Their Diclectin doses were subsequently increased according to body weight and individual symptoms. A follow up phone call occurred within 1 to 3 weeks after the intervention, at which time the overall PUQE score was repeated, along with individual scoring of symptoms of nausea, vomiting, and retching. RESULTS: Sixty-eight women were enrolled and completed the study. Despite moderate to severe NVP, defined by the validated PUQE scoring system, most women (50/68) were receiving 2 tablets a day of Diclectin instead of the recommended dose of 4 tablets a day. Following a mean doubling of the dose to 4 tablets a day, there was a significant decrease in length of nausea (from 4 to 3 hours, P < 0.001), frequency of vomiting (from mean 1.6 to 1.3 a day, P = 0.02), and overall PUQE score (from mean 7.5 to 6.1, P < 0.001). CONCLUSION: Women suffering from NVP are often given subtherapeutic doses of Diclectin. Women should receive a dosage according to their body weight and severity of their symptoms. PMID- 14532952 TI - The TGF-beta superfamily and its roles in the human ovary and placenta. AB - The transforming growth factor-beta (TGF-beta) superfamily consists of a large group of growth and differentiation factors, such as TGF-betas, activins, inhibins, growth and differentiation factors (GDFs), and bone morphogenetic proteins (BMPs). These molecules act through specific receptor complexes that are composed of type I and type II serine/threonine receptor kinases. The receptor kinases subsequently activate Smad proteins, which then propagate the signals into the nucleus to regulate target gene expression. Several ligands in this family, such as TGF-betas, activins, inhibins, BMP-15, and GDF-9, play important roles in regulating human ovarian functions, including follicle development and maturation. Activin and TGF-beta are also involved in regulating placental development and functions. Abnormal expression or function of these ligands has been found in several pathological conditions. This review summarizes the role of the TGF-beta superfamily in human ovarian and placental regulation and function, and the potential clinical implications. PMID- 14532953 TI - Vaginal birth after caesarean section: clinical and legal perspectives. AB - Vaginal birth after Caesarean section (VBAC) is currently the preferred method of delivery for pregnant women who have undergone 1 previous low transverse Caesarean section. This common practice warrants some reconsideration in light of recent clinical data on the risks associated with VBAC, and it is incumbent upon clinicians to ensure that women under their care are fully aware of these risks. Indeed, in some circumstances, an attempt at VBAC may be perceived by the courts to represent a negligent standard of care. PMID- 14532954 TI - The "glitazones": rosiglitazone and pioglitazone. AB - Rosiglitazone and pioglitazone are two new additions to the therapeutic options for the treatment of type 2 diabetes mellitus. These agents differ from our current therapies in their mode of action. They have potential non-glucose lowering effects that may reduce cardiovascular risk and are effective both as monotherapy and in combination with sulfonylureas, metformin, and insulin. They are generally well tolerated, with the main side effects being weight gain and fluid retention. However, special precaution is warranted in patients with congestive heart failure or hepatic disease, and monitoring of liver enzymes is recommended for the first year of therapy. Despite their effectiveness, rosiglitazone and pioglitazone remain second-line agents to metformin and glyburide, agents that have demonstrated efficacy in decreasing the microvascular and macrovascular complications associated with type 2 diabetes mellitus. PMID- 14532955 TI - Medical management of a cervical pregnancy: a case report. AB - BACKGROUND: Cervical pregnancy is a rare condition that can become life threatening if heavy bleeding occurs. CASE: Grace (pseudonym) is a 32-year-old woman who was admitted to hospital at approximately 6 weeks' gestational age after several days of heavy bleeding. Ultrasonographic examination revealed the presence of cervical pregnancy. She received multiple doses of methotrexate with folinic acid rescue, and did not require surgical intervention. Nine months after the resolution of the cervical pregnancy, Grace again became pregnant. She had an uncomplicated gestation and a normal vaginal delivery of a healthy baby at term. CONCLUSION: Multiple-dose methotrexate with folinic acid rescue was a safe option for the medical treatment of ectopic cervical pregnancy in this case. Long-term toxicity studies and case reports are required to provide more information regarding safety and for clinicians to individualize treatment regimens. PMID- 14532956 TI - The use of first trimester ultrasound. AB - OBJECTIVE: To review the clinical indications for first trimester ultrasound. OUTCOME: Proven clinical benefit from first trimester ultrasound. EVIDENCE: MEDLINE search and bibliography reviews in relevant literature. VALUES: Content and recommendations reviewed by the principal authors and the Diagnostic Imaging Committee of the Society of Obstetricians and Gynaecologists of Canada. Levels of evidence were judged as outlined by the Canadian Task Force on the Periodic Health Examination. RECOMMENDATIONS: 1. First trimester ultrasound is recommended for assessment of threatened abortion to document fetal viability (II-2B) or for incomplete abortion to identify retained products of conception (II-2B). 2. First trimester ultrasound is not recommended to diagnose pregnancy, to date pregnancy when last normal menstrual period and physical examination are concordant, or to investigate an inevitable abortion (II-2B). First trimester ultrasound is indicated when last menstrual period date is uncertain (I-A). 3. First trimester ultrasound is recommended prior to pregnancy termination. (II-2B) 4. First trimester ultrasound is recommended during diagnostic or therapeutic procedures requiring visual guidance (e.g., chorionic villus sampling, amniocentesis) and prior to cervical cerclage placement. (I-A) 5. First trimester ultrasound is recommended for suspected multiple gestation to allow for reliable determination of chorionicity or amnionicity. (II-2A) 6. First trimester ultrasound is recommended for suspected ectopic pregnancy, molar pregnancy, and suspected pelvic masses. (II-1A) 7. First trimester ultrasound is recommended for early assessment of anatomic development in situations of increased risk for major fetal congenital malformations. (II-3C) 8. Nuchal translucency screening should only be offered as part of a comprehensive prenatal screening and counselling program by experienced operators with appropriate quality assurance processes in place (II-2A). PMID- 14532957 TI - Recent developments in cardiac pacing. PMID- 14532958 TI - Balloon angioplasty of native coarctation of the aorta in a local group of children: acute results and midterm angiographic re-assessment. AB - OBJECTIVE: Balloon angioplasty of native coarctation of the aorta is gaining acceptance as an alternative to surgery in children. The aim of this study was to assess the acute and midterm effectiveness and safety of the procedure. METHODS: During a 3-year period, nine patients with native coarctation underwent balloon angioplasty at a median age of 2.9 years (range: 4 mo - 12 y) and median weight of 11 kg (range: 3.1 - 51 kg). Balloon diameter selected was at least twice the diameter of the narrowed segment, but never more than the diameter of the aorta at diaphragmatic level. RESULTS: There was a significant improvement in the mean systolic gradient across the coarctation from 36 +/- 13mm Hg to 4 +/- 4 mm Hg (p < 0.001), and in the mean diameter of the narrowed segment from 4 +/- 1.3 mm to 7.2 +/- 2.7 mm (p < 0.001). There was no immediate mortality or complication. Follow-up catheterisation was done in eight patients, a mean of 2.7 years (range: 0.8 - 4 y) after the initial procedure. No evidence of restenosis was found, but one patient developed an aneurysm that was surgically resected. CONCLUSION: Balloon angioplasty is effective and safe for the treatment of native coarctation in children. Radiographic follow-up is advisable in the midterm. Balloon angioplasty offers an alternative to surgery in the treatment of selected children older than 3 months with native coarctation of the aorta. PMID- 14532959 TI - Prosthetic valve obstruction at Tygerberg Hospital between January 1991 and February 2001. AB - BACKGROUND: Prosthetic valve obstruction is a relatively rare, but potentially fatal complication in patients with prosthetic heart valves. The diagnosis and appropriate management of these patients present a challenge to both the cardiologist and the cardiac surgeon. Despite efforts over the last 30 years to prevent this complication, it remains a lifelong risk. Obstruction is caused by pannus formation, thrombus formation or a combination of pannus and thrombus. Valve replacement has traditionally been the treatment of choice. METHODS: Patients were selected from echocardiography and surgical reports between January 1991 and February 2001. All patients were analysed with regard to demographic information, clinical features, imaging results, treatment and outcome data. INR values on presentation were obtained from hematology archives. RESULTS: A total of 32 patients presented on 34 occasions. There were 25 women and seven men. Obstruction occurred in the mitral position in 56% of cases and in the aortic position in 44% of cases. All but two valves were St Jude bileaflet valves. Patients generally presented with sever dyspnoea (NYHA class IV in 64.7%) and poor anticoagulation control (INR < 2.5 in 75.8% ). The initial imaging modality used in all cases was transthoracic echocardiography. Fluoroscopy was used in five cases transesophageal echocardiography in only two cases. Valve replacement was performed on 20 patients, six patients received thrombolysis and the remaining eight patients did not receive any treatment. Outcome was poor with an overall mortality of 643.7%. CONCLUSION: Given the extremely high mortality rate with current management, the treatment of prosthetic valve obstruction with thrombolysis in selected patients deserves consideration in a prospective study. PMID- 14532960 TI - Mitral valve prolapse: a study of 45 children. AB - The knowledge pertaining to mitral valve prolapse is mainly based on studies in adults. In this study, the clinical profile as described in adults was compared with that found in children up to the age of 13 years. Forty-five children with echocardiographic-proven mitral valve prolapse and who met the inclusion criteria were included in the study. The male:female ratio in this study was 1:1.37 and was not statistically significantly different from reported ratios. Most of the children were asymptomatic. Twenty-one of the 31 patients referred from outside the hospital had an incidentally found murmur. The symptoms found in this study were not similar to those described in adults. The most commonly found symptoms were shortness of breath and fatigue, in contrast to those of chest pain and palpitations described in adults. Comparing males to females in this study, significantly lower weight (p = 0.005) and body mass index (p = 0.003) were found in girls, and a significantly lower pulse rate (p = 0.002) in boys. Left-sided cardiac enlargement was diagnosed in 11 patients on chest X-ray and in six patients on electrocardiogram. One patient had Marfan syndrome and four others had a Marfanoid appearance. In conclusion, most children with mitral valve prolapse are asymptomatic. Mitral valve prolapse is not an uncommon finding in children younger than 13 years of age. Patients with mitral valve regurgitation were advised to take infective endocarditis prophylaxis prior to invasive procedures. PMID- 14532961 TI - Ability of Nigerian hypertensive patients to perceive changes in their blood pressure. AB - The objective of this study was to examine the ability of hypertensive patients to predict correctly when their blood pressure (BP) was elevated, and the symptoms they felt when such changes occurred. We also sought to determine whether literacy levels or the number of years that a patient had been diagnosed as hypertensive had a correlation with whether a patient was a predictor and whether he was an accurate predictor. One hundred and seventy-seven patients were studies, of which 101 (57.1%) claimed they could tell when their BP was elevated. Only 45 (44.5%) of the predictors were right in their prediction. Although being literate and longer time of being hypertensive was found to be likely to make a patient a predictor, neither of these variables was likely to make a predictor accurate. Predictors were also likely to be younger than non-predictors. The symptoms the patients were likely to complain about as denoting a rise in BP included headache (20.4%), weakness/tiredness (20.4%) and palpitation (18.5%). The study has shown that the majority of predictors were not accurate and so hypertensive patients should be counseled not to depend on how they feel but to visit physicians regularly for BP checks. PMID- 14532962 TI - Is there a role for 99mTc-HMPAO leucocyte scintigraphy in infective endocarditis? AB - Infective endocarditis is an important disease in developing countries. A combination of clinical findings, typical organisms cultured from blood, and specific echocardiographic features are used to establish the diagnosis. The diagnosis is often difficult because cultures are not always positive and transthoracic echocardiography lacks sensitivity and specificity, leading to delayed treatment or inappropriate therapy. As (99m)Tc-HMPAO-labeled leucocyte scintigraphy is used routinely to evaluate patients with suspected infectious processes, it was postulated that this technique might also aid in the diagnosis of infective endocarditis in patients with underlying chronic rheumatic heart disease or other valvular disease. Six patients were referred for (99m)Tc-HMPAO labeled leucocyte scintigraphy. The white blood cells were labeled according to standard procedures. Whole-body planar imaging and single photon emission tomography of the chest area, with imaging at 30 minutes, 3 hours and 24 hours after the administration of the labeled leucocytes, were performed on all patients. All the scintigrams were negative. The reasons for the negative findings are not entirely clear, but probably reflect the pathological nature of vegetations, which consist mainly of masses of fibrin, clotted platelets and blood cell debris, containing the causative organisms. Leucocytes do not play a major role in the pathological process. Although only six patients were studied, it appears that (99m)Tc-HMPAO leucocyte scintigraphy is of little value in the evaluation of patients with infective endocarditis. A study after the administration of radiolabelled antibiotics may be of greater value and should be considered in these patients. PMID- 14532963 TI - Separation of parental genomes in human blood and bone marrow cells and its implications. AB - The linear order of genes is apparently interrupted at chromosomal ends. Our observations on human blood and bone marrow cells indicate that the chromosomes of each of the two parental sets maintain coherence, perhaps in tandem, forming a ring. Two such rings in a diploid cell join building a larger ring, which folds up to form the interphase nucleus. The linear order of genes thus extends beyond the chromosomal ends. These observations become especially significant when seen in the light of cell biologic findings on interaction of chromosomes or chromatin and centrioles in different cell cycle phases, in polymorphonuclear cells and during the zygotic developments. They may explain how the genomic order and the sequential continuity of the genes are maintained and why such order remains often cryptic. PMID- 14532964 TI - Reduced expression of the regulatory A subunit of serine/threonine protein phosphatase 2A in human breast cancer MCF-7 cells. AB - The beta1 subunit of integrin is serine/threonine phosphorylated in growth arrested human breast cancer MCF-7 cells, while it is not in quiescent normal human breast epithelial (HBE) cells. Using the affinity-purified antibodies PB788 9 against the synthetic oligopeptide that contained phosphothreonines corresponding to threonines 788 and 789 on beta1 integrin, beta1 integrin in MCF 7 cells, but not in HBE cells, was found to react with PB788-9. The beta1 integrin immunoprecipitates from HBE cells co-immunoprecipitated the core enzyme of serine/threonine protein phosphatase (PP) 2A, consisting of the regulatory A (PP2A-A) and the catalytic C (PP2A-C) subunits, with the protein phosphatase activity susceptible to okadaic acid (OA), an inhibitor of PP2A and PP1, but not to a PP1 inhibitor. In contrast, beta1 integrin from MCF-7 cells co immunoprecipitated PP2A-C, but not PP2A-A, with no protein phosphatase activity. Immunoblotting of whole cell lysates revealed that a comparable amount of PP2A-C was present in either HBE or MCF-7 cells, but the amount of PP2A-A was significantly reduced in MCF-7 cells compared to that in HBE cells. The results suggest that the failure of beta1 integrin dephosphorylation at threonines 788 and 789 may be due to a significant reduction in the PP2A-A expression in MCF-7 cells. PMID- 14532965 TI - The early response of p53-dependent proteins during radiotherapy in human rectal carcinoma and in adjacent normal tissue. AB - The aim of this study was to investigate the activation of the p53 pathway and the induction of apoptosis during preoperative radiotherapy in normal human rectal tissue and in rectal carcinoma. Twelve patients with rectal cancer of the lower third were enrolled in this study. Tumor specimens and adjacent normal tissue were obtained before radiation, after the third radiation cycle and from the surgically removed rectum. All specimens were analyzed be means of immunohistochemistry for expression of p53 and its downstream target genes MDM2 and p21. In normal mucosal crypts, irradiation led to p53 accumulation and MDM2 induction in more than 70% of the cells. The accumulation of p53 in basal crypts was associated with high expression of p21. Apoptosis was also induced in crypts and occurred in 15% of the cells. Activation of the p53 pathway was not seen in the resting cells at the luminal border of the epithelium. In interstitial cells, p21 was highly upregulated, whereas p53 and MDM2 showed weak expression. The level of bcl-2 was not altered during radiotherapy in healthy tissue. In rectal carcinoma cells, p53 expression was unaltered by irradiation in 11 out of 12 tumors. The p53 non-functional tumors were characterized by a weak induction of MDM2 and p21 and by the lack of apoptosis in the presence of bcl-2. Our findings demonstrate that sequential immunohistochemical analysis is suitable to detect a deregulation of the p53 pathway in human rectal cancer cells during radiotherapy. Further investigations are necessary to elucidate its value as a prognostic marker and potential predictor of therapy responsiveness. PMID- 14532966 TI - Aspirin inhibits highly invasive prostate cancer cells. AB - Cell adhesion, proteolytic degradation and cell migration are interrelated processes responsible for the invasion and metastasis of cancer. One of the crucial molecules involved in cancer metastasis is urokinase-type plasminogen activator (uPA). An elevated concentration of uPA is a strong indicator of poor prognosis. In addition to the proteolytic activity of uPA, which degrades the extracellular matrix, uPA also binds to its receptor (uPAR) and controls cell adhesion and migration through the reorganization of actin cytoskeleton. We have recently demonstrated that constitutively active nuclear factor-kappa B (NF kappaB) is responsible for the increased secretion of uPA and that inhibition of NF-kappaB suppresses secretion of uPA and cell migration of highly invasive cancer cells. Aspirin and other nonsteroidal anti-inflammatory drugs have been recently shown to have a chemopreventive effect in colon and pancreatic cancers. Here we show that aspirin inhibits NF-kappaB, resulting in the suppression of uPA secretion from the highly invasive human prostate cancer cells PC-3. Furthermore, aspirin inhibited migration of PC-3 cells, suggesting an effect on the uPA-uPAR signaling complex. Finally, aspirin suppressed adhesion of PC-3 cells to fibronectin (FN), which binds to an alpha3beta1 integrin receptor, and to vitronectin (VN), which binds to alphavbeta3 integrin receptor. Altogether, our data suggests that aspirin inhibits the formation of uPA-uPAR-FN-alpha3beta1 and uPA-uPAR-VN-alphavbeta3 complexes, resulting in the suppression of cell adhesion and cell motility of the highly invasive prostate cancer cells PC-3. These results indicate that aspirin may contribute directly to reducing invasion and metastasis of prostate cancers by inhibiting cell migration and invasion. PMID- 14532967 TI - Relationship between HER2 status and proliferation rate in breast cancer assessed by immunohistochemistry, fluorescence in situ hybridisation and standardised AgNOR analysis. AB - Lack of standardisation and inaccuracy in HER2 test results may adversely influence patient evaluation and therapy selection. In the present study we applied immunohistochemistry (IHC) using the A0485 and CB11 antibodies and fluorescence in situ hybridisation (FISH) for detection of HER2 in 74 routinely processed breast carcinoma specimens. The rapidity of cellular proliferation was assessed by standardised AgNOR analysis and compared with HER2 status. Protein over-expression was found in 30/74 cases by A0485 and in 20/74 by CB11 antibodies, while amplification was detected in 22/74 carcinomas by FISH. Twenty seven of 74 tumours were high-level AgNOR expressors (mean AgNOR area >3.369 microm2), 19 of which revealed amplification. The highest concordance between results was achieved by FISH and CB11-IHC (97%), the concordance between FISH and A0485-IHC was 89 and 84% between FISH and AgNOR quantity, respectively. The overall concordance between A0485 and CB11-IHC was 85% with 10 incongruent cases, all scored 2+ by A0485 and 0/1+ by CB11. Eight of the discordant tumours were non amplified by FISH and 7 were low AgNOR-expressors. Our results indicate that using CB11 antibody, a nearly complete agreement can be achieved between HER2 IHC and FISH in diagnostic paraffin material. Moreover, in 2+ positive IHC cases, the AgNOR analysis may represent an additional tool to select patients as candidates for Herceptin therapy due to the strong negative predictor value. PMID- 14532968 TI - Establishment and characterization of novel xenograft models of human biliary tract carcinomas. AB - In order to develop new therapeutic regimens for biliary tract cancers, which carry dismal prognoses, the establishment of a human biliary tract cancer xenograft model is essential. Herein, we report the successful establishment and characterization of two xenograft models of human biliary tract cancers. An adenosquamous gallbladder cancer cell line (TGBC-44) and a bile duct adenocarcinoma cell line (TGBC-47) were obtained from fresh surgical specimens in our department and subcutaneously inoculated into nude mice. The overall tumor take rate was 100% and solid tumors grew measurable after 5 and 7 days for TGBC 44 and TGBC-47, respectively. Tumor doubling time was 3.9+/-1.1 and 4.1+/-0.5 days in the exponential growth phase in TGBC-44 and TGBC-47 xenografts, respectively. Isozyme test and karyotype analysis confirmed the human origin. Histopathology analysis revealed that the TGBC-44 xenograft retained both the squamous and the adenocarcinoma components, and the TGBC-47 xenograft exhibited poorly differentiated adenocarcinoma as in the corresponding original tumors. Immunohistochemistry and Western blotting studies revealed positive and similar expression of platelet derived endothelial growth factor/thymidine phosphorylase (PDGF/TP), thymidylate synthase (TS), and cyclooxygenase-2 (COX-2) in both original tumors and xenograft models. No macroscopic metastases were found at the time of sacrifice. We have successfully established two models of human biliary tract cancer, gallbladder and bile duct cancer. Models retained the morphological and biochemical characteristics of the original tumor and demonstrated constant biological behavior in all transplanted mice. These models could be useful tools for developing new diagnostic and therapeutic strategies against biliary tract cancers. PMID- 14532969 TI - Accumulation of Hsp70/Hsc70 molecular chaperone regulator BAG-1 on COPI-coated structures in gastric epithelial cells. AB - The multifunctional protein BAG-1 binds to molecular chaperones Hsc70/Hsp70 and regulates their activity. While a role for BAG-1 in regulating Hsc70 chaperone activity is firmly established in vitro, physiological roles of this regulation remain obscure. Here we show that BAG-1 is highly expressed in gastrointestinal epithelial cells and accumulates at the Golgi apparatus, probably through interaction with COPI coated structure. Subcellular fractionation of gastric mucosa revealed that BAG-1 was mainly recovered in the Golgi-enriched fractions. Confocal immunofluorescence studies revealed that BAG-1 was overlapped with a cis Golgi matrix protein GM130 and a COPI component beta-COP in the vicinity of the Golgi. In response to brefeldin A, which blocks recruitment of COPI coats to the Golgi membrane, BAG-1 was dispersed throughout the cytoplasm as beta-COP was. Extensive overlap of BAG-1 immunofluorescence with beta-COP was still observed when cells were treated with nocodazole, which depolymerizes microtubules and induces fragmentation of the Golgi stacks. Immunoelectron microscopy revealed that BAG-1 signals were predominantly found on vesicular membrane structures adjacent to Golgi stacks, but not on mitochondrial membrane. Taken together, we suggest that BAG-1 is targeted to the COPI coated structures, implying its contribution toward the COPI vesicular transport in gastrointestinal epithelial cells. PMID- 14532970 TI - Bystander effect in suicide gene therapy is directly proportional to the degree of gap junctional intercellular communication in esophageal cancer. AB - Gap junctional intercellular communication (GJIC) has been shown to be involved in the bystander effect through herpes simplex virus thymidine kinase/ganciclovir (HSV-tk/GCV) gene therapy. In this study, we examined the expression of connexins, the components of gap junction, and the degree of GJIC in esophageal cancer cell lines and compared the bystander effect in cells with different capacities of GJIC. We found loss in connexin 26 expression and reduced connexin 43 in esophageal cancer. GJIC capacity varied among cell lines and was dependent on the connexin 43 expression in the cell-cell contact areas. In mixing assay, the extent of the bystander effect was tightly correlated with the degree of GJIC capacity. The effects of retinoic acid and cAMP on the bystander effect were also investigated. Treatment with retinoic acid, but not with cAMP, was associated with augmented bystander killing by increase in GJIC in some esophageal cancer cell lines. Our results indicated that the degree of GJIC was predictive to identify a tumor as suitable for gene therapy with the HSV-tk/GCV system. Also GJIC chemically-enhanced with retinoic acid might be useful to improve response in suicide gene therapy. PMID- 14532971 TI - Cyclooxygenase-2 upregulates vascular endothelial growth factor expression and angiogenesis in human gastric carcinoma. AB - Although gastric cancer with cyclooxygenase (COX)-2 overexpression is associated with poor prognosis, the mechanistic pathway remains unknown. We examined the associations between expressions of COX-2 and vascular endothelial growth factor (VEGF) in both gastric cancer cells and in human gastric cancer. The gastric cell line, Kato III, was transiently transfected with cox-2 expressing vector. The levels of COX-2, prostaglandin (PG) E2 and VEGF expression were measured post transfection. Additionally, expressions of COX-2 and VEGF in human gastric cancer were determined by immunohistochemistry in archive gastrectomy specimens. Tumor angiogenesis was assessed by the microvessel density (MVD), which was determined by anti-CD34 immunostaining. Transient transfection of Kato III with cox-2 was associated with increased COX-2 expression, higher PGE2 production and upregulated VEGF expressions. Treatment with NS398, a specific COX-2 inhibitor, reduced VEGF expression in COX-2 expressing Kato III cells by 25%. Among the 67 gastric cancers examined, COX-2 overexpression was found in 45 (67%) cases whereas increased VEGF expression was detected in 46 (69%) cases. There was a significant association between COX-2 and VEGF expressions in gastric cancer (r=0.25, p=0.041). Additionally, tumor MVD was associated with both COX-2 (r=0.32, p=0.008) and VEGF (r=0.39, p=0.001) expressions. Our results showed that overexpression of COX-2 in both gastric cells and primary gastric cancer is associated with upregulation of VEGF and angiogenesis. Future studies should evaluate the potential anti-angiogenic effect of COX-2 inhibitors on human gastric cancer. PMID- 14532972 TI - Proteome analysis identified maspin as a special feature of papillary thyroid carcinoma. AB - This study aimed at investigating new mechanisms of carcinogenesis in thyroid cancer at the molecular level and at finding potential protein markers involved in the initiation of the different histological subtypes. For this, we performed differential proteome analysis on primary cultured thyrocytes (PT) and transformed thyrocytes (TT) derived from 238Pu alpha-particle irradiation using 2 dimensional electrophoresis (2-DE) and peptide mass fingerprinting (PMF) with matrix-assisted laser desorption/ionisation-time of flight mass spectrometry (MALDI-TOF MS). Image analysis showed that one protein was very strongly expressed in TT; 55 proteins were weaker, different in intensity, including 26 spots that were increased in PT, and 29 spots were decreased. The hot spot was identified as maspin, a unique member of the serpin family considered to be a class II tumor suppressor gene. To clarify the role of maspin in thyroid carcinogenesis we searched for protein expression in 20 normal (tumor-free) tissues, as well as in 20 follicular adenomas (FAD), 20 papillary carcinomas (PTC), 20 follicular carcinomas (FTC), 20 poorly differentiated carcinomas (PDTC), and 20 undifferentiated carcinomas (UTC). Maspin protein expression was detectable in none of the cases of normal tumor-free thyroid tissue, nor in FAD, FTC, PDTC and UTC. In contrast 14 of 20 PTC (70%) showed a moderate or strong cytoplasmic staining; 4 of these 14 cases had a moderate cytoplasmic and nuclear staining. In conclusion, we hypothesize that maspin protein expression is a special feature in the cascade of PTC genesis and that the way of initiating PTC is different from other thyroid carcinoma types. PMID- 14532973 TI - Targeting foreign major histocompatibility complex molecules to tumors by tumor cell specific single chain antibody (scFv). AB - Down-regulation of the major histocompatibility complex (MHC) is one of the major mechanisms that tumor cells adopted to escape immunosurveillance. Therefore, specifically coating tumor cells with foreign MHC may make tumor cells a better target for immune recognition and surveillance. In this study, we designed and generated a fusion protein, H2Kd/scPSMA, consisting of a single chain antibody against human prostate specific membrane antigen (PSMA) and the extracellular domain of mouse H-2Kd. The expression of this fusion protein in B16F0 mouse melanoma cells was confirmed by RT-PCR and fluorescent activated cell sorting (FACS). Our animal study showed that the expression of H2Kd/scPSMA in B16F0/PSMA5, a B16F0 cell line expressing human PSMA, significantly inhibited tumor growth as demonstrated in the pulmonary metastasis assay and tumor growth study and improved overall survival. PMID- 14532974 TI - The modifier subunit of glutamate cysteine ligase (GCLM) is a molecular target for amelioration of cisplatin resistance in lung cancer. AB - It is unclear whether a subunit of glutamate cysteine ligase [a modifier subunit (GCLM) and a catalytic subunit (GCLC)] is an effective target for ameliorating cisplatin (CDDP)-resistance. We inhibited each subunit of GCL mRNA using a specific ribozyme (M-Rz and C-Rz) in the pulmonary adenocarcinoma cell line A549. GCL activity was suppressed by the ribozyme. CDDP-resistance was more effectively ameliorated when GCLM rather than GCLC was inhibited. GCLM is a potentially more effective pharmacologic target for ameliorating CDDP-resistance in non-small cell lung cancer than GCLC. PMID- 14532975 TI - Macrophages play a critical role in the anti-tumor activity of Virulizin. AB - Virulizin, a novel biological response modifier (BRM), has been demonstrated to have a high level of anti-tumor activity against pancreatic cancer and melanoma in many clinical trials and preclinical studies. However, its anti-tumor mechanism has not been fully elucidated. The purpose of this study was to define the mechanism of Virulizin anti-tumor activity in cultures and in a murine xenograft model. The presence of Virulizin stimulated in a dose-dependent manner the cytolytic activity against tumor cells by splenocytes and macrophages, but not by non-adherent splenocytes. The cytotoxic activity of macrophages was significantly increased (approximately 5-fold) in cultures containing 2.5% of Virulizin compared to that of cultures without Virulizin (p<0.001). An increase of 21% in the protease secretion was observed in Virulizin (2.5%)-stimulated macrophages compared to PBS-treated cells (p<0.0001). Moreover, the anti-tumor efficacy of Virulizin observed in CD-1 nude mice was abrogated in mice that were depleted of macrophages, thus stimulation of macrophages may be one mechanism through which Virulizin acts. These results suggest that macrophages may play a critical role in the anti-tumor activity of Virulizin. PMID- 14532976 TI - Enhanced antitumor activity of irofulven in combination with 5-fluorouracil and cisplatin in human colon and ovarian carcinoma cells. AB - Irofulven (6-hydroxymethylacylfulvene, MGI-114, NSC 683863) is a semisynthetic derivative of illudin S, a natural product obtained from the Omphalotus mushroom. Irofulven has demonstrated potent activity against a broad range of solid tumors in both cellular and xenograft models and has shown promising activity in clinical trials. To guide the clinical use of irofulven, the present study used the MTT viability assay to examine the cytotoxic effects obtained by combining irofulven with two other anticancer agents: cisplatin and 5-fluorouracil (5-FU). The study was carried out with HT-29 and HCT-116 colorectal and A2780 ovarian carcinoma cells as well as with their irofulven- (HT-29/IF2, HCT-116/IF27) or cisplatin-resistant (A2780/CP70) variants. The combinations showed strong sequence specificity. Simultaneous exposure to cisplatin and irofulven was at least additive for four cell lines including the cisplatin-resistant A2780/CP70 ovarian cells which exhibit a multifactorial resistance phenotype. Cisplatin followed by irofulven was additive for parental HCT-116 and A2780 cells whereas irofulven followed by cisplatin was antagonistic in all cellular models. Simultaneous exposure to 5-FU and irofulven was at least additive for all six cell lines. 5-FU followed by irofulven was additive for the parental HT-29 and A2780 cells and synergistic for the irofulven-resistant HCT-116 cell line. Irofulven followed by 5-FU was synergistic for the two ovarian cell lines and additive for the two parental colon cell lines. These studies demonstrate that simultaneous exposure to irofulven and cisplatin is at least additive for most cell lines whereas simultaneous exposure to irofulven and 5-FU is additive to synergistic for all the cell lines tested, including the irofulven- and cisplatin resistant variants. The enhanced cytotoxicity of irofulven in combination with cisplatin and 5-FU support the clinical application of these regimens. PMID- 14532977 TI - Microsatellite instability at chromosome 8p in non-small cell lung cancer is associated with lymph node metastasis and squamous differentiation. AB - Genetic alterations at chromosome arm 8p are associated with advanced disease and poor patient outcome in several types of malignant tumors. We studied the frequency of microsatellite instability (MSI) and loss of heterozygosity (LOH) at chromosome 8p in early stage non-small cell lung cancer (NSCLC) of 47 patients with stage I or II disease (25 squamous cell carcinomas and 22 adenocarcinomas). Microsatellite analysis was performed after laser microdissection using 5 polymorphic tetranucleotide microsatellite markers and 4 dinucleotide markers at chromosome 8p. A pentanucleotide repeat marker at the chromosomal locus 17p13.1 (TP53.Alu) was also analyzed. Expression of the mismatch repair (MMR) proteins hMSH2, hMSH6 and hMLH1 was evaluated by immunohistochemistry. Microsatellite instability (MSI) in at least 2 markers was detected in 9 of 47 patients (19.1%) and was predominantly found at tetranucleotide repeats. Sixteen of 47 (34.0%) NSCLC demonstrated LOH at chromosome 8p. All MSI-positive tumors showed normal expression of the MMR proteins. The presence of MSI at chromosome 8p was associated with lymph node metastasis (p=0.02), squamous differentiation (8/25; 32%-p=0.03), and the presence of LOH at the p53 locus (p=0.06). None of the other investigated clinical, pathologic or molecular factors correlated with MSI. Our study showed that an elevated MSI at selected tetranucleotide sequences (EMAST) on chromosome 8p is frequent in early stage squamous cell carcinomas of the lung with lymphatic spread. The tetranucleotide marker panel used in this study was able to indicate lymph node metastasis and high risk disease in patients with resectable squamous cell lung cancer. PMID- 14532978 TI - Expression of ADAMs (a disintegrin and metalloproteases) and TIMP-3 (tissue inhibitor of metalloproteinase-3) in human prostatic adenocarcinomas. AB - A disintegrin and metalloproteases (ADAMs) are proteins that contain both a disintegrin and metalloprotease domain and have potential implications for the metastasis of human cancer cells via cell adhesion and protease activities. In this study, we analyzed the expression levels of ADAM-9, ADAM-10 and ADAM-17 (TNF alpha converting enzyme, TACE), and TIMP-3 (tissue inhibitor of metalloproteinase 3) in human prostatic tumor cell lines as well as in clinical patient materials (BPH and tumor tissue samples). Human prostatic tumor cell lines (MDA PCa 2b, LNCaP-C33, -C51, -C81, -Pro5, -Ln3, -C4-2, PC3, and DU145) showed varied levels of expression for ADAM-9, -10 and -17 mRNA. A strong expression of ADAM-17/TACE was further revealed by Western blot analysis in prostatic tumor cell lines. In the case of clinical material, all the tumor samples (8/8) revealed the expression of ADAM-9, -10, and -17 compared to the specimens of benign prostatic hyperplasia (BPH) where 80% of (8/10) samples showed the expression of ADAM-9, 86% (6/7) of ADAM-10, and 30% (3/10) of ADAM-17/TACE. Interestingly, expression of a potent inhibitor of ADAM-17 (TIMP-3) was not detected in any prostatic tumor cell lines (0/9), while TIMP-3 expression was detected in 82% (9/11) of BPH samples. Androgen-sensitive LNCaP-C33 cells exhibited differences in ADAMs regulation by 5alpha-dihydrotestosterone (DHT), while such differences were not detectable in androgen-independent LNCaP-C81 cells. These results suggest that an inverse expression pattern of ADAM-17/TACE and TIMP-3, and the regulation of ADAMs with DHT might play an important role in the pathogenesis of prostate cancer. PMID- 14532979 TI - Down-regulation of TNF-alpha receptors by conophylline in human T-cell leukemia cells. AB - In the course of our screening for tumor necrosis factor-alpha (TNF-alpha) function inhibitors, conophylline, a vinca alkaloid isolated from the plant Ervatamia microphylla, was found to inhibit TNF-alpha-induced NF-kappaB activation. We studied the effect of conophylline on TNF-alpha-induced NF-kappaB and JNK activations in human T-cell leukemia Jurkat cells. Conophylline inhibited both of these TNF-alpha-induced activations. It also inhibited phosphorylation and degradation of I-kappaB-alpha. Moreover, a receptor binding assay using [125I]-TNF-alpha showed that this inhibitory effect was due to a decrease in the binding of TNF-alpha to the cells. Scatchard analysis of the binding data indicated that conophylline induced only a small change in the affinity of the receptors but a significant change in the receptor number. FACS analysis showed that conophylline reduced the expression of CD120a/TNFR1, the high-affinity receptor for TNF-alpha, on the cell surface. On the other hand, conophylline did not affect the kinetics of internalization and degradation of TNF-alpha/receptor complexes or the half-life of TNF-alpha binding sites. These results indicate that conophylline down-regulates the expression of the TNF-alpha receptors on the cell surface. PMID- 14532980 TI - HER2 testing in gastric cancer: molecular morphology and storage time-related changes in archival samples. AB - Seventy-two archival specimens of advanced gastric adenocarcinoma were analyzed by fluorescence in situ hybridization and immunohistochemistry for HER2 gene status. Gene amplification and concomitant protein overexpression were detected in 15.2% of cases. Intratumor topographical variability of HER2 amplification was observed, indicative of the late involvement of HER gene in gastric tumorigenesis. No significant correlations were found between HER2 status and histopathologic type, grade of differentiation, and Goseki classification of carcinomas, suggesting that HER2 gene is variably linked with the morphogenesis of gastric adenocarcinoma. A significant decrease of the incidence of HER2 amplified carcinomas in function of the duration of the storage of paraffin blocks was observed: 42-33% for tumor specimens paraffin-embedded in 2002-2001; 8.3%, 15.7%, 11.1% for the years 2000, 1999, 1998, respectively; 0% for cases embedded during 1997. According to these results, the reliability of the FISH and immunohistochemical assays decreases after prolonged storage of paraffin-embedded specimens. PMID- 14532981 TI - Clinical significance of telomerase activity in basal cell carcinomas and in tumour-free surgical margins. AB - To distinguish the infiltrative from the non-infiltrative basal cell carcinoma (BCC), cell cycle markers are being used to supplement histopathological assessment, and proliferation markers are proving particularly useful. A successful radical therapeutic intervention depends on a clear histopathological diagnosis, especially for the tumour margins. For this purpose we investigated whether proof of telomerase activation is a suitable adjunctive molecular marker. We were also interested in the telomerase activity (TA) of BCC-free margin tissues as a prognostic parameter of relapse. Using PCR-ELISA kits, we found TA in 26/30 (87%) BCC tissues and in 8/25 (32%) of the tumour-free surgical margin tissues. Telomerase levels and the incidence of telomerase-positive tumour tissues are not always associated with positive p53 immunoreactive scores in BCC tissues. But telomerase levels correlate significantly with p53 expression levels. In the Kaplan-Meier curve, patients with telomerase-positive tumour-free surgical margin tissues showed significantly shorter relapse-free periods than patients with telomerase-negative tumour-free margin tissues. PMID- 14532982 TI - Hypericin-mediated photodynamic therapy induces lipid peroxidation and necrosis in nasopharyngeal cancer. AB - Photoactivation of hypericin is known to generate singlet oxygen and superoxide anion radicals. Reactive oxygen species (ROS) produced by photodynamic therapy (PDT) has the capacity to induce oxidative damage and tumor destruction. We have previously shown that hypericin-PDT induces tumor shrinkage and regression in the human nasopharyngeal cancer (NPC)/HK1 murine tumor model. In this extended study, we show by electron microscopy that subcutaneously implanted HK1 NPC cells from Balb/c nude mice perished by cell necrosis with hypericin-PDT treatment. There was evidence of cytoplasmic swelling accompanied by loss of cell membrane integrity and autophagic vacuolization of cytoplasm but no nuclear changes. There was also no significant difference in the apoptotic index of control and PDT treated tumors, when analyzed by in situ end labeling of DNA strand breakage to detect apoptosis. This further supports the observation that cell death in PDT treated NPC/HK1 tumors was by necrosis. Lipid peroxidative stress analyzed by the malonaldehyde assay was significantly elevated in PDT-treated cells. However, PDT had no effect on the activity of superoxide dismutase, an intracellular antioxidant enzyme. The findings show that hypericin-PDT of nasopharyngeal tumors in vivo induces tumor necrosis with accompanying lipid peroxidation. PMID- 14532983 TI - Black cohosh, a menopausal remedy, does not have estrogenic activity and does not promote breast cancer cell growth. AB - Black cohosh is an increasingly popular alternative to estrogen replacement therapy for the relief of menopausal symptoms, primarily hot flushes. However, an important consideration for long-term therapy is potential toxicity and carcinogenicity. Therefore, we undertook a study to assess the estrogenic activity of black cohosh to examine its safety for those with, or at high risk of developing, breast cancer. Several assays were utilized as listed: RNAse protection assays, which ascertain the regulation of the expression of E2 responsive genes; estrogen-responsive-element (ERE)-luciferase, which determines modulation of the ER function by transactivation of the ERE; the Ishikawa cell system, which has an E2-regulated endogenous alkaline phosphatase; and colony formation of ER-expressing breast cancer cells, which indicates possible progression of early stage breast cancer into a more aggressive state. Black cohosh extracts did not demonstrate estrogenic activity in any of these assay systems. This is an encouraging step in the assessment of the safety of black cohosh for treatment of menopausal hot flushes. PMID- 14532984 TI - Growth inhibitory and apoptotic effects of inositol hexaphosphate in transgenic adenocarcinoma of mouse prostate (TRAMP-C1) cells. AB - Inositol hexaphosphate (IP6), a dietary agent, has been extensively studied for its cancer chemopreventive and anti-cancer efficacy in several different animal models, but not in prostate cancer (PCA) possibly because the known PCA models are both expensive and highly time-consuming. One such PCA model is transgenic adenocarcinoma of mouse prostate (TRAMP), which reproduces the spectrum of benign latent, aggressive and metastatic forms of human PCA. In this study, we investigated the anti-cancer effects of IP6 in TRAMP-derived established TRAMP-C1 cell line. IP6 (0.5-4.0 mM) treatment of cells for 24-72 h resulted in 17-76% cell growth inhibition and 6-35% cell death, in a dose- and time-dependent manner. In the studies assessing whether cell growth inhibition by IP6 is associated with an alteration in cell cycle progression, IP6 treatment resulted in up to 92% cells in G0-G1 phase as compared to controls. In other studies assessing its apoptotic efficacy, IP6 induced a moderate to strong (up to 14-fold over control) apoptotic cell death. In additional studies, pretreatment of cells with all caspases inhibitor for 2 h followed by 2 mM IP6 for 48 h resulted in approximately 50% reversal in IP6-induced apoptosis suggesting a partial involvement of caspases activation in IP6-caused apoptosis. Furthermore, IP6 showed significant induction (6-fold) in caspase-3 activity compared to control suggesting the involvement of caspases activation in IP6-induced apoptosis. However, pretreatment of cells with all caspase inhibitor, which partially reversed the IP6-induced apoptosis, completely inhibited IP6-induced caspase-3 activation, providing convincing evidence of both caspase-dependent and independent mechanisms in IP6-induced apoptotic death of TRAMP-C1 cells. Together, these results warrant further mechanistic and in vivo efficacy studies with IP6 in TRAMP and other pre-clinical models of PCA. PMID- 14532985 TI - Quantitative molecular diagnosis of peritoneal lavage fluid for prediction of peritoneal recurrence in gastric cancer. AB - We developed a quantitative multiple-marker RT-PCR assay for sensitive detection of free cancer cells in the peritoneal cavity and examined the significance of this molecular diagnostic technique for detection and prediction of peritoneal dissemination in patients with gastric cancer. Preoperative peritoneal lavage fluid samples obtained from 129 patients with gastric cancer were subjected to RT PCR assay with primers specific for carcinoembryonic antigen (CEA) and cytokeratin-20 (CK-20), and conventional cytological examination with Papanicolaou staining. The multi-marker RT-PCR assay was positive in 59 of 129 (46%) gastric cancer patients, whereas conventional cytology was positive in only 9 of 129 (7%) patients. Thirty-two of 129 (22%) patients suffered disease recurrence after surgery. Twenty-one of these patients were confirmed to have had peritoneal recurrence. Although conventional cytology was positive on peritoneal washes in only 9 patients, the RT-PCR assay was positive in 20 of these 21 patients. Furthermore, in cases with negative cytology, patients with PCR positive findings in peritoneal lavage fluid had a significantly poorer prognosis than those with negative PCR, mainly because of peritoneal recurrence. Our results suggest that the multiplex RT-PCR assay for CEA and CK-20 was highly sensitive for detection and might be useful for prediction of peritoneal dissemination in gastric cancer. PMID- 14532986 TI - Differentially expressed genes and estrogen receptor status in breast cancer. AB - There is a large and increasing body of experimental and clinical data supporting the involvement of estrogen on the proliferation of hormone-dependent breast tumors. Estrogen acts via its receptor (ER) stimulating cellular proliferation. ER and progesterone receptor (PR), which is regulated by estrogen via ER, have been used as prognostic markers in the clinical management of breast cancer patients. The aim of the present study was the identification of tumor-associated genes differentially expressed in breast tumors regarding the presence or absence of ER and PR. Using the technique of differential display reverse transcriptase polymerase chain reaction (DDRT-PCR) we have isolated and cloned 127 cDNA fragments that showed differential expression in either ER+/PR+ or ER-/PR- breast tumors. Sequencing analysis of these clones revealed that 119 cDNAs had homology with known sequences in the National Center of Biotechnology Information (NCBI) and 8 were novel, showing no homology to known genes. Among these differentially expressed transcripts are metabolic enzymes, ribosomal proteins, transcription factors, hypothetical proteins, cell cycle regulators, cytoskelectum related genes, cell adhesion and motility genes. Differences in gene expression profiles are likely to explain the phenotypic differences between hormone-responsive and hormone-unresponsive breast tumors. PMID- 14532987 TI - p53-deficient cells display increased sensitivity to anthracyclines after loss of the catalytic subunit of the DNA-dependent protein kinase. AB - DNA-dependent protein kinase (DNA-PK) is involved in non-homologous end joining which repairs DNA double-strand breaks introduced by irradiation and radiomimetic agents. DNA-PK interacts with p53 but may also have p53-independent functions. The present study investigated whether disruption of the gene for the catalytic subunit DNA-PKcs affects chemosensitivity in p53-deficient cells. Drug sensitivity of DNA-PKcs(+/+)/p53(+/+), DNA-PKcs(+/+)/p53(-/-), DNA-PKcs(-/ )/p53(+/+), and DNA-PKcs(-/-)/p53(-/-) mouse lung-fibroblasts was determined by the MTT assay, the clonogenic assay, and trypan blue exclusion. Susceptibility to apoptosis was determined by DNA fragmentation (TUNEL) and by caspase-3 cleavage. We show that p53-deficient cells were 2 to 3-fold resistant to treatment with doxorubicin, epirubicin, cisplatin, and docetaxel as compared to wild-type cells. We further demonstrate that the additional loss of DNA-PKcs function in p53 deficient cells resulted in a 2-fold increase in sensitivity to doxorubicin and epirubicin as documented by the MTT assay, clonogenic assay, and trypan blue exclusion. Doxorubicin-induced hypersensitivity in these cells correlated with a transient G2/M checkpoint activation but did not seem to correlate with apoptosis. The data indicate that additional loss of DNA-PKcs in p53-deficient cells reverses anthracycline-resistance imposed by p53-deficiency, and that DNA PKcs modulates p53-independent pathways responding to DNA damage induced by anthracyclines. They also indicate that processes other than apoptosis may contribute to the increased cytotoxicity to anthracyclines. DNA-PKcs may thus be a potential target for functional inhibition, which might increase the efficacy of some anti-tumour agents in the treatment of cancers mutated in the p53 gene. PMID- 14532988 TI - Bcl-2 expression is a prognostic factor in the subgroups of patients with colorectal cancer. AB - The prognostic significance of Bcl-2 expression in colorectal cancer has been intensively studied, however, the results were controversial in the whole group of colorectal cancer patients. We proposed that one of the main reasons for such controversial results may be that Bcl-2 played variable roles in the subgroup of patients. We, therefore, investigated the prognostic importance of Bcl-2 expression by using immunohistochemistry in the various subgroups of 147 patients with colorectal cancer. Among these tumours, 85 (58%) expressed Bcl-2 protein and 62 (42%) were negative. Bcl-2 expression was positively related to DCC expression (p=0.0002). Survival analyses in the subgroups of the patients showed that lack of Bcl-2 expression was related to a worse prognosis in the male patients (p=0.02) but not in female patients (p=0.53), in the patients with DNA diploid tumours (p=0.005) not in the patients with non-diploid tumours (p=0.46), and in the patients with ras negative tumours (p=0.01) not in the patients with ras positive tumours (p=0.25). Bcl-2 expression was not related to prognosis in the total group of the patients (p=0.20). In conclusion, Bcl-2 protein may play variable prognostic roles in the subgroups of the patients with colorectal cancer. Analysis of Bcl-2 expression in the tumour may be of value in predicting prognosis and therapeutic response. PMID- 14532989 TI - Photodynamic therapy of transitional cell carcinoma multicellular tumor spheroids with hypericin. AB - The objective of the present study was to evaluate the sensitivity of the bladder transitional cell carcinoma (TCC) to hypericin PDT in a 3-D system using multicellular tumor spheroids. The photodynamic response in RT-112 human bladder TCC spheroids was also compared to 2-D cultured monolayer cells. Following a 2-4 h incubation with 8-30 microM hypericin, spheroids or monolayer cells were irradiated at the light dose of 12 J/cm2, delivered at a fluence rate of 10-100 mW/cm2. The PDT effects were evaluated using a clonogenic assay. The results show that compared with the cells in a monolayer, cells in spheroids were dramatically less sensitive to hypericin PDT (<2000-fold). Studies of fluorescence microphotographs of centrally cut frozen sections of hypericin-exposed spheroids showed a gradient in hypericin concentration from the peripheral to the central region of the spheroid. Although it can be suggested that heterogeneity of drug uptake might be responsible for the observed resistance of spheroid to hypericin PDT, hypericin sensitized spheroids that were dissociated prior to light irradiation were as sensitive as the monolayer cells to hypericin PDT, suggesting that other factors such as oxygen depletion might be responsible for the resistance of spheroids to hypericin PDT. PMID- 14532990 TI - Telomerase in relation to expression of p53, c-Myc and estrogen receptor in ovarian tumours. AB - Telomerase activity and its subunits (hTERC, hTERT mRNA) were evaluated in ovarian tumours in relation to the expression of p53, c-Myc and estrogen receptor (ER). Furthermore, relations between telomerase activity, hTERC and hTERT with known clinicopathologic prognostic factors and survival in patients with malignant tumours was investigated. Telomerase activity was determined with TRAP, hTERC and hTERT with RT-PCR, while p53, c-Myc and ER expression with immunohistochemistry. Telomerase activity and hTERT mRNA were more frequently observed in malignant ovarian tumours compared to borderline and benign tumours, whereas hTERC was present in all tumour types. p53 and c-Myc were more frequently detected in malignant compared to borderline and benign tumours. Telomerase activity was positively related to hTERT mRNA, p53 and c-Myc expression, but not to hTERC and ER expression. In malignant tumours, hTERC levels were related to tumour stage, while telomerase activity and hTERT mRNA expression were not related to any clinicopathologic feature. Tumour stage, differentiation grade, residual tumour after first laparotomy and presence of ascites were related to (progression free) survival, whereas telomerase activity or its subunits were not. In conclusion, these data suggest that p53 expression (e.g. p53 mutation) as well as c-Myc expression may have a role in regulation of telomerase activity in ovarian tumours. PMID- 14532991 TI - PC-SPES: Molecular mechanism to induce apoptosis and down-regulate expression of PSA in LNCaP human prostate cancer cells. AB - PC-SPES is an eight-herbal mixture which has activity against prostate cancer cells and can reduce the serum level of prostate specific antigen (PSA) in more than 80% of individuals with prostate cancer. We conducted this study to begin to clarify the molecular mechanism by which PC-SPES inhibited the growth of prostate cancer cells and down-regulated expression of PSA. Western blot analysis, luciferase reporter assay using a variety of promoters of the PSA gene and the isolated androgen receptor response elements (ARE), as well as electrophoretic mobility shift assay (EMSA) were employed to study the effect of PC-SPES on DHT induced expression of PSA in LNCaP androgen-dependent human prostate cancer cells. Also, Western blot analysis and luciferase reporter assay using 12-0 tetradecanoylphorbol-13-acetate response elements were employed to study the ability of PC-SPES to activate the c-Jun NH2-terminal kinase (JNK)/c-Jun/AP-1 signal pathway in these cells. Reporter studies showed that PC-SPES inhibited DHT induced PSA promoter/enhancer-luciferase activity via inhibition of ARE transcriptional activity. Western blot analysis showed that PC-SPES down regulated DHT-induced expression of PSA without decreasing DHT-induced nuclear level of AR. EMSA demonstrated that PC-SPES inhibited the binding of DHT activated AR to ARE. Moreover, we found that PC-SPES phosphorylated JNK, increased levels of phosphorylated and unphosphorylated forms of c-Jun, and enhanced AP-1 transcriptional activity in LNCaP cells. Interestingly, when LNCaP cells were stably tranfected with the dominant negative JNK binding domain (JBD) of JNK-interacting protein-1 (JIP-1), these cells no longer underwent apoptosis and growth inhibition in the presence of PC-SPES. But, PC-SPES still decreased levels of PSA in the LNCaP-JIP-1 cells. Taken together, PC-SPES inhibited binding of DHT-activated AR to AREs of PSA gene resulting in down-regulation of ARE transcriptional activity and expression of PSA, and this occurred independently of the JNK/c-Jun/AP-1 signal pathway. Also, PC-SPES activated the JNK/c-Jun/AP-1 signal pathway resulting in growth arrest and apoptosis of prostate cancer cells. PMID- 14532992 TI - Identification and characterization of human KIAA1391 and mouse Kiaa1391 genes encoding novel RhoGAP family proteins with RA domain and ANXL repeats. AB - The commonly deleted region of breast cancer at human chromosome 11q23.1 is located between microsatellite markers D11S927 and D11S1347. We have previously identified and characterized the KIAA1735 gene, encoding MTH and DIX domain protein, within the 11q23.1 commonly deleted region. The BTG4 gene, encoding BTG/TOB family protein, as well as the SIK2 gene, encoding salt-inducible serine/threonine kinase 2, were also located within the 11q23.1 region. We identified and characterized the KIAA1391 gene within the 11q23.1 commonly deleted region by using bioinformatics. The nucleotide position 11-3586 of human KIAA1391 cDNA was predicted as the coding region of human KIAA1391 gene. Mouse Kiaa1391 gene was located within mouse genome sequences RP23-95E13 (AC102770.4), RP23-212M11 (AC140363.1), and mouse chromosome 9 genomic contig NT_039473.1. Exon intron structure was well conserved between human KIAA1391 gene and mouse Kiaa1391 gene. Nucleotide sequence of mouse Kiaa1391 cDNA was determined in silico by assembling nucleotide sequences of exon 1-15 of mouse Kiaa1391 gene. Human KIAA1391 protein (1191 aa) and mouse Kiaa1391 protein (1182 aa) showed 79.1% total-amino-acid identity. RA domain (codon 198-274), RhoGAP domain (codon 363-546), and two Annexin-like (ANXL) repeats (codon 657-767 and 796-909) of human KIAA1391 protein were conserved in mouse Kiaa1391 protein. This is the first report on the comprehensive characterization of human KIAA1391 gene and mouse Kiaa1391 gene, encoding RhoGAP proteins with RA domain and two ANXL repeats. PMID- 14532993 TI - Identification and characterization of human LL5A gene and mouse Ll5a gene in silico. AB - ARCN1, KIAA0638, TREH, DDX6, BLR1, BCL9L, UPK2, DLNB13, DLNB14, RPS25, SBDN, G6PT1, HYOU1, VPS11, HMBS, H2AFX, DPAGT1, KIAA0285, MIZF, ABCG4, NOD9, PDZK2, CBL, MCAM, RNF26, C1QTNF5, MFRP, USP2, THY1, and PVRL1 genes are located within the commonly deleted region of neuroblastoma at human chromosome 11q23.3. Here, we characterized the KIAA0638 gene within the 11q23.3 region by using bioinformatics. Because human KIAA0638 gene was homologous to human LL5B gene encoding LL5beta protein, KIAA0638 was designated LL5A gene encoding LL5alpha protein. LL5A isoform 1 (FLJ00141 type) consists of exons 1-12, 14-21 and 23, while LL5A isoform 2 (KIAA0638 type) consists of exon 1-23. LL5A isoform 1 was the major transcript among LL5A isoforms generated due to alternative splicing. Nucleotide sequence of mouse Ll5a cDNA was determined by assembling CB522359 EST and 5'-truncated mKIAA0638 cDNA. Human LL5alpha isoform 2 showed 94.4 and 35.9% total-amino-acid identity with mouse Ll5alpha and human LL5beta, respectively. LL5alpha proteins consist of Forkhead associated (FHA) domain, bipartite nuclear localization signal (NLS), Chromosome segregation ATPases (SMC) domain, and Pleckstrin homology (PH) domain. LL5alpha proteins were homologous to PtdIns(3,4,5)P3 sensor protein LL5beta in the SMC and PH domains, and were also homologous to KIF1A, KIF1B, KIF13A, KIF13B, KIF14, and SNX23 proteins in the FHA domain. LL5alpha protein might be a transducer of PtdIns(3,4,5)P3 levels to the intracellular trafficking system. PMID- 14532994 TI - Delivery of recombinant adeno-associated virus by jet injection. AB - The jet-injection technology was used for delivery of recombinant adeno associated virus (rAAV). Although AAV-based vectors are an attractive tool in gene therapy, some methodological and technical problems of their targeted delivery remain to be solved. We tried to address some of these cell-targeting problems by using a new low-volume needleless injection device the Swiss Injector. First we tested, by electron microscopy, whether jet-injection would have any detrimental effect on rAAV particle integrity. Second, we compared transgene expression after infection of 293T cells with fired or control (non fired) rAAV that expressed the green fluorescent protein (GFP), beta galactosidase (beta-gal), the B7.1 molecule, and interleukin 2 (IL2). Third, an rAAV carrying the genes coding for beta-gal was jet-injected into mouse subcutaneous (s.c.) tumours. The staining of tumour cryosections revealed beta gal expression 72 h after the delivery. Our study demonstrated the applicability of the Swiss Injector for the delivery of rAAV into tumour tissue without either vector particle integrity or the level of expression of the transgenes, as tested in vitro, being affected. The jet-injection technology could improve the distribution of vector particles in the tumour mass without leakage of liquid from the injection site. PMID- 14532996 TI - Treatment of ulcerative colitis patients by long-term administration of germinated barley foodstuff: multi-center open trial. AB - Germinated barley foodstuff (GBF), which mainly consists of dietary fiber and glutamine-rich protein, is a prebiotic for ulcerative colitis (UC). In our previous study, we carried out a clinical trial of GBF with mildly to moderately active UC patients and showed that GBF treatment was able to attenuate the symptoms of UC in a relatively short-term. The aim of this study was to investigate the efficacy of long-term administration of GBF in the treatment of UC in a multi-center open trial. Twenty-one patients with mildly to moderately active UC received 20-30 g of GBF for 24 weeks in an open-label protocol while baseline treatments (5-amino-salicyrate compounds and/or steroids) were continued. The response to the GBF treatment was evaluated using a clinical scoring and after 24 weeks of observation, the GBF group showed a significant decrease in clinical activity index (especially, the degree of visible blood in stools and the presence of nocturnal diarrhea) compared with the control group (p<0.05). No side effects related to GBF were observed. In conclusion, GBF can reduce the clinical activity of UC over long-term as well as short-term administration. Nutraceutical GBF therapy may have a place in long-term management of UC, but controlled studies are needed to demonstrate its efficacy in the treatment of this disorder. PMID- 14532995 TI - E-cadherin-dependent intercellular adhesion enhances chemoresistance. AB - E-cadherin, an intercellular adhesion molecule, is important in cell growth and differentiation. Adhesion between cells is thought to decrease as cancers develop and disseminate. Knowledge of the effect of cell adhesion on proliferation and chemosensitivity may help individualize cancer treatment. Lovo and MCF-7 cells, which express E-cadherin, and PC-3 cells, which do not, were used in this study. Proliferation and chemosensitivity were measured in two-dimensional (2-D) culture and three-dimensional (3-D) culture. Protein and mRNA expression of E-cadherin, catenin, and cyclin-dependent kinase inhibitors were determined. Growth of Lovo and MCF-7 but not PC-3 cells was markedly suppressed in 3-D relative to 2-D. MCF 7 cells express high levels for E-cadherin, catenin, and p27 in 3-D, but catenin and p27 expression was decreased by exposure to anti-E-cadherin neutralizing antibody. Chemosensitivity of PC-3 was similar in 2-D and 3-D, but chemosensitivity of Lovo and MCF-7 was less in 3-D than 2-D. Moreover, the presence of anti-E-cadherin antibody increased chemosensitivity of MCF-7 in 3-D. E-cadherin affected the regulation of cell proliferation and differentiation, and decreased chemosensitivity. Chemosensitivity of cancer is affected by the state of cell adhesion and expression of intercellular adhesion molecules. Consideration of intercellular adherence characteristics in different chemosensitivity tests is likely to improve their reliability. PMID- 14532997 TI - Overexpression of X-linked inhibitor of apoptosis in human hepatocellular carcinoma. AB - The X-linked inhibitor of apoptosis (XIAP) is a member of a novel family of inhibitors of apoptosis. Since suppression of apoptosis is fundamentally important for carcinogenesis and tumor growth, we investigated the expression and function of XIAP in human hepatocellular carcinomas (HCCs). XIAP was expressed constitutively in HCC cell lines. Fourteen out of 20 (70%) HCC tissues demonstrated moderate or strong cytoplasmic staining for XIAP, whereas non-tumor parts showed negative or weak staining for XIAP by immunohistochemistry. In addition, XIAP expression was inversely correlated with apoptosis, but not with proliferation in HCC tissues. These results indicated that XIAP is a principal inhibitor of apoptosis overexpressed in human HCCs and that XIAP may be a potential target for gene therapy of human HCCs. PMID- 14532998 TI - Overexpression of cyclooxygenase-2 protein and its relationship to apoptosis in cervical carcinoma treated with neoadjuvant chemotherapy. AB - The aims of this study were to examine the overexpression of COX-2 protein and its relationship to apoptosis in cervical carcinoma patients treated with neoadjuvant chemo-therapy (NAC), and to assess the potential role of COX-2 as a predictor of the response to NAC in a series of patients with cervical carcinoma. For immunohistochemical analysis, cervical cancer tissue samples were collected before NAC and 3 weeks after NAC using transcatheter arterial infusion of cisplatin from 40 patients who underwent surgery for advanced cervical carcinoma in stages IB, IIA and IIB and from 5 normal cervical tissues between 1991 and 2000 at the Department of Obstetrics and Gynecology, under informed consent. Patients were randomly assigned to receive one or two arterial infusions of cisplatin. COX-2 protein expression was detected by immunohistochemical staining and was classified as no expression for tumors with negative or <10%, while > or =10% positive staining was defined as overexpression. Detection of apoptosis was done by the TUNEL method. The percentage of cells with DNA fragmentation (apoptotic index, AI) was calculated before NAC and 3 weeks after NAC. The AI ratio (AI after NAC/AI before NAC) was also calculated. COX-2 expression was not detected in the normal cervix. Overexpression of COX-2 protein was detected in 18 out of 40 (45.0%) cervical cancers. A higher incidence of COX-2 protein overexpression was observed in patients with adenocarcinoma than in those with squamous cell carcinoma (p=0.1797, Fisher's exact text). The average AI value before and after NAC was 8.85 versus 11.82 respectively. In COX-2 protein negative patients with squamous cell carcinoma, the AI ratio was 0.96+/-0.46 following one arterial infusion of cisplatin and 3.19+/-2.72 following two infusions of cisplatin. There was a significant positive correlation between apoptosis and the number of infusions of cisplatin (p=0.0098, Mann-Whitney, U test). Our findings suggest that COX-2 protein expression could be used as a predictor of chemoresistance and that assessment of the COX-2 status could be useful to identify cervical cancer patients who may benefit from NAC. PMID- 14532999 TI - Differential expression of homing receptor CD103 on lamina propria lymphocytes and association of CD103 with epithelial adhesion molecules in inflammatory bowel disease. AB - Intraepithelial lymphocytes play an important role in mucosal immunology, and are involved in the pathogenesis of inflammatory bowel disease. We studied expression of CD103 on mucosal lymphocytes with epithelial adhesion molecules in patients with inflammatory bowel disease. Surgical specimens of human colon were obtained from 12 patients with ulcerative colitis, 12 patients with Crohn's disease, and 5 controls. Frozen sections were cut and expression of CD103 on lymphocytes, E cadherin, CD44V3, and CD44v6 on intestinal epithelium was studied. Frequency of CD103-positive intraepithelial lymphocytes did not differ among controls, patients with ulcerative colitis, and patients with Crohn's disease. The frequency of CD103-positive lamina propria lymphocytes was significantly higher in patients with Crohn's disease than in controls and patients with ulcerative colitis. The frequency of CD103-positive intraepithelial lymphocytes was significantly correlated with that of lamina propria lymphocytes in patients with ulcerative colitis. The frequency of CD103-positive intraepithelial lymphocytes was significantly correlated with epithelial E-cadherin expression but that of lamina propria lymphocytes was not. Differential up-regulation of CD103 expression on lamina propria lymphocytes in Crohn's disease may indicate differential humoral or cellular regulation in inducing CD103 molecules on lymphocytes in patients with this disease. PMID- 14533000 TI - N-3 fatty acid-rich diet prevents early response of interleukin-6 elevation in trinitrobenzene sulfonic acid-induced enteritis. AB - Dietary fat is an important factor involved in the pathogenesis of inflammatory bowel disease (IBD). It remains unclear how n-3 and n-6 fatty acids modulate intestinal inflammation. In this study, we investigated the effects of n-3 and n 6 fatty acid-rich diets on trinitrobenzene sulfonic acid (TNBS)-induced enteritis. The rats were fed an n-3 or n-6 rich-diet for 12 days, and then starved for the following 2 days. An intraileal injection of TNBS was administered, and TNBS-enteritis subsequently developed. Macroscopic and histological examination was performed after 24 h. Serum cytokine levels were determined by ELISA. The n-6 fatty acid-rich diet markedly enhanced mucosal damage as compared to the n-3 fatty acid-rich diet. The damage score was significantly higher in the n-6 fatty acid-rich diet group (P<0.05). Histological changes in the mucosa were more severe in the n-6 fatty acid-rich diet group than in the n-3 fatty acid-rich group. Serum IL-6 levels were significantly higher in the n-6 fatty acid-rich diet group than in the n-3 fatty acid-rich group (P<0.05). On the other hand, there were no differences in serum TNF-alpha levels. The n-3 fatty acid-rich diet effectively reduced early mucosal inflammation in TNBS enteritis. The effects of the n-3 fatty acids were associated with blockage of mucosal IL-6 secretion. Our data suggest that n-3 fatty acid-rich diet may be applicable for enteral nutrition in the treatment of IBD patients. PMID- 14533001 TI - 11q23-24 loss is associated with chromosomal instability in endometrial cancer. AB - A loss of the DNA copy number at chromosomal region 11q23-24 as detected by comparative genomic hybridization (CGH) is a marker of poor prognosis in patients with endometrial cancer. Malignant tumors display genetic instability, which is classified into two types: microsatellite instability (MIN) and chromosomal instability (CIN). In the present study, we examined whether there is a relation between loss of 11q23-24 and genetic instability in endometrial adenocarcinoma. Loss of 11q23-24 was detected in 4 of 70 endometrial cancers by fluorescence in situ hybridization (FISH), and DNA aneuploidy was detected by laser scanning cytometry (LSC) in 14 tumors. All tumors with 11q23-24 loss were aneuploid, and three of them were considered to have CIN. These findings suggest that 11q23-24 contains gene(s) necessary for normal chromosome replication and cell division. PMID- 14533002 TI - Induction of apoptosis by Se-MSC in U937 human leukemia cells through release of cytochrome c and activation of caspases and PKC-delta: mutual regulation between caspases and PKC-delta via a positive feedback mechanism. AB - Se-methylselenocysteine (Se-MSC) has been shown to possess potent chemopreventive and anti-tumor properties. However, its exact mechanism of action is still not well understood. The present study investigated the mechanism of Se-MSC on the induction of apoptosis using U937 human leukemia cells. Se-MSC induced dose- and time-dependent apoptosis of U937 cells as assessed by flow cytometry analysis, DNA fragmentation, and proteolytic cleavage of poly-(ADP-ribose) polymerase (PARP). Se-MSC increased time- and dose-dependent cytochrome c accumulation in the cytosol, which was greatly inhibited by overexpression of Bcl-2, suggesting that the apoptotic effect by Se-MSC in U937 cells is mitochondrial-dependent. Se MSC also induced activation of caspases, followed by proteolytic cleavage of PKC delta. The Se-MSC-induced apoptosis required activities of caspases since pretreatment of a pan-caspase inhibitor z-VAD-fmk greatly suppressed the Se-MSC induced apoptosis as well as proteolytic cleavage of PKC-delta, suggesting activation of caspases is critical for the Se-MSC-induced apoptosis, and caspases lie upstream of PKC-delta. The Se-MSC-induced apoptosis of U937 cells also required activity of PKC-delta because pretreatment of rottlerin, a specific PKC delta inhibitor greatly blocked the Se-MSC-induced apoptosis as well as processing and activities of caspases, suggesting activation of PKC-delta is also important for the Se-MSC-induced apoptosis of U937 cells, and PKC-delta lies upstream of caspases. Together, our data suggest the apoptotic mechanism by Se MSC in U937 cells may be related to cytochrome c release from the mitochondria, and mutual activation between caspases and PKC-delta via a positive feedback mechanism, which may potentiate the apoptotic action by Se-MSC in U937 cells. PMID- 14533003 TI - Mechanism of thermotolerance induction by split-dose hyperthermia in Deinococcus radiodurans DNA repair deficient mutants. AB - We examined the phenomenon of thermotolerance induction in the radioresistant prokaryote, Deinococcus radiodurans, which was initially exposed to 30 min at 52 degrees C followed by various intervals up to 6 h at 30 degrees C in TGY medium and then re-exposed to 52 degrees C for various periods, i.e., split-dose hyperthermia. This thermotolerance induction was analyzed in DNA repair deficient mutants (strain 302, 251, UVS25, rec30 and KH840) and the wild-type strain MR1. The strain UVS25 is a double mutant for the mtcA and uvsD genes, and strain rec30 is a mutant for the deinococcal recA gene. The induction was suppressed to 1/10 and 1/25 in strains UVS25 and rec30 respectively, as compared with the maximum level in the wild-type strain MR1. However, the induction in strain 302 (mutant for the uvrA gene) was not suppressed. Therefore, we conclude that proteins synthesized during the interexposure interval, i.e., the products of the uvsD (UV endonuclease beta) and recA (RecA protein) genes contribute to the induction of thermotolerance in D. radiodurans. PMID- 14533004 TI - Genotypes of CCR2 and CCR5 chemokine receptors in human myasthenia gravis. AB - The aim of this study was to examine the association of human autoimmune myasthenia gravis (MG) with two DNA polymorphisms of the chemokine receptors CCR5 Delta 32 and CCR2-64I. CCR2 and CCR5 interact primarily with the human CC family ligands CCL2 (formerly called monocyte chemoattractant protein; MCP-1), CCL3 and CCL4 (macrophage inflammatory protein-1 alpha and -1 beta; MIP-1 alpha/beta), and their main function is to recruit leukocytes from circulation into the tissues, thus playing an important role in human inflammatory disorders. A PCR-based genotyping method was used to determine the genetic variation at the CCR5 gene and an automated real-time Pyrosequencing technology was employed for the analysis of G right curved arrow A point mutation at the CCR2 gene. Results obtained from 158 patients and 272 healthy controls demonstrate no evidence of association between genetic variants of CCR2 and CCR5 with MG and its clinical manifestations. CCR2-64I and CCR5-Delta 32 genotypes are thus unlikely to be involved in protection or predisposition to MG. PMID- 14533005 TI - Preventive effect of zinc acexamate administration in streptozotocin-diabetic rats: Restoration of bone loss. AB - The preventive effect of zinc compounds on bone loss in streptozotocin (STZ) diabetic rats was investigated. Rats received a single subcutaneous administration of STZ (6.0 mg/100 g body weight), and 7, 14 or 21 days later the animals were sacrificed by bleeding. STZ administration caused a significant decrease in body weight and a significant increase in serum glucose and triglyceride levels, indicating diabetic condition. Femoral-diaphyseal and metaphyseal alkaline phosphatase activity, calcium and deoxyribonucleic acid (DNA) contents were significantly decreased by STZ administration, showing that diabetic condition causes bone loss. Zinc sulfate (2.5 mg Zn/100 g) or zinc acexamate (2.5 mg Zn/100 g) was orally administered once daily for 14 days to rats received a single subcutaneous administration of STZ (6.0 mg/100 g). STZ administration-induced increase in serum glucose and triglyceride levels and decrease in body weight, femoral-diaphyseal and -metaphyseal alkaline phosphatase activity, DNA and calcium contents were significantly prevented by the administration of zinc acexamate. The preventive effect of zinc sulfate on bone components was not seen. The present results demonstrate that the administration of zinc acexamate has a preventive effect on bone loss in STZ-diabetic rats in vivo. PMID- 14533006 TI - Expression and localization of ecto-nucleotide pyrophosphatase/phosphodiesterase I-3 (E-NPP3/CD203c/PD-I beta/B10/gp130RB13-6) in human colon carcinoma. AB - Ecto-nucleotide pyrophosphatase/phospho-diesterase-I enzyme (E-NPP), one of the type II transmembrane proteins, cleaves phosphodiester and phosphosulfate bonds of a variety of substrates including deoxynucleotides, NAD, and nucleotide sugars. Mammalian E-NPP consists of three closely related family proteins; E-NPP1 (PC-1), E-NPP2 (PDNP2/PD-Ialpha/autotaxin), and E-NPP3 (CD203c/PDNP3/PD Ibeta/B10/gp130RB13-6) that express in different cells or at different locations even in the same cell. E-NPP3 is associated with malignant subversion and invasive properties. In this study, the expression and localization of E-NPP3 were investigated in human colon carcinoma. Western blotting showed strong E-NPP3 expression in cancer tissues and in the serum of colon carcinoma patients. Immunohistochemically, E-NPP3 was expressed not only in the apical but also in the basolateral plasma membranes of cancer cells. No prominent pattern of intracellular localization, and no relation between clinical stage and E-NPP3 expression were observed. Our results suggested that E-NPP3 is associated with carcinogenesis of human colon cancer and that serum E-NPP3 might be a tumor marker of colon carcinoma. PMID- 14533007 TI - RB1CC1 suppresses cell cycle progression through RB1 expression in human neoplastic cells. AB - RB1-inducible Coiled-Coil 1 (RB1CC1) is a putative transcription factor that functions as a key regulator of retinoblastoma 1 (RB1). RB1CC1 mutations lacking this function are involved in the tumorigenesis of breast cancers. RB1CC1 is distributed in various tissues other than the breast, and is thought to play a biological role in controlling cell growth and progression of various cancers. The present study examined the correlation between RB1CC1 and cell cycle-related molecules in human neoplastic cells, and the ratios of cells at various phases of the cell cycle were verified in the RB1CC1-transduced human leukemic cell lines, K562 and Jurkat. The results showed that RB1CC1 was synchronously expressed with RB1 in various cell lines and that introducing RB1CC1 induced RB1 expression in human leukemic cell lines, although independently of the other molecules. Western blotting showed that underphosphorylated forms of RB1 were elicited by RB1CC1, whereas E2F1 was not affected. Cell cycle analysis demonstrated that G2-M phases were suppressed in RB1CC1-transduced cells. These data suggested that RB1CC1 induces the expression of RB1, especially of underphosphorylated forms, then suppresses cell cycle progression in human neoplastic cells. PMID- 14533008 TI - Molecular detection of free cancer cells in pleural lavage fluid from esophageal cancer patients. AB - The clinical significance of free cancer cells in pleural lavage fluid detected by molecular methods during surgery remains uncertain in esophageal squamous cell cancer (ESCC). We therefore evaluated the relationship between free cancer cells and clinicopathological findings, and compared the reverse transcriptase polymerase chain reaction (RT-PCR) method with conventional cytological examination. Pleural lavage fluid from 38 consecutive patients was obtained at two time points; immediately after thoracotomy and before thorax closure. Papanicolaou and Giemsa staining as well as carcinoembryonic antigen (CEA) specific RT-PCR were performed. The positivity rates obtained using cytological examination and CEA-mRNA expression were 5.3 and 15.8%, respectively. Positive results were observed in pleural lavage fluid after tumor resection. No significant differences in clinicopathologic factors were seen, irrespective of CEA-mRNA expression status. Among the 5 patients exhibiting CEA-mRNA positivity, 2 experienced hematogenous recurrence, 2 experienced mixed recurrence and 1 experienced pleural dissemination. With regard to mode of recurrence and mean period between surgery and relapse, no significant differences were seen between CEA-mRNA-positive and CEA-mRNA-negative patients. Although disease recurred in almost all patients exhibiting CEA-mRNA expression, due to the relatively small sample in the present study the clinical significance must be investigated further in a larger number of patients. PMID- 14533009 TI - Ghrelin clearance is reduced at the late stage of polymicrobial sepsis. AB - The cardiovascular response to sepsis is characterized by an early, hyperdynamic phase followed by a late, hypodynamic phase. Ghrelin, a newly-identified endogenous ligand for growth hormone secretagogue receptor (i.e., ghrelin receptor), was recently demonstrated to be a potent vasoactive peptide in addition to its effects on growth hormone release and energy homeostasis. We have shown that ghrelin (via its receptor) may play an important role in regulating cardiovascular responses in the progression of polymicrobial sepsis. However, it remains unknown whether the clearance of this peptide is altered in sepsis. To determine this, male adult rats were injected with 125I-ghrelin through the jugular vein at 5 or 20 h after cecal ligation and puncture (CLP, i.e., sepsis model) or sham operation. The blood sample was collected every 2 min for 30 min for determining half-life (t1/2). Tissue samples (i.e., kidneys, liver, brain, heart, lungs, spleen, stomach, small intestine, large intestine, skin and muscle) were then harvested. The radioactivities of samples were counted. The results indicate that 125I-ghrelin's t1/2 and its distribution were not significantly altered in early sepsis (5 h after CLP). However, the t1/2 increased significantly in late sepsis (20 h after CLP). Tissue distribution of 125I ghrelin was far greater in the kidneys than in any other tissues tested in both sham and septic animals. Moreover, the kidneys and liver had significantly less radioactive uptake at 20 h after CLP, but the radioactivity in blood was much higher at the same time point. There were no significant changes in 125I-ghrelin distribution in other organs at the late stage of sepsis. These results indicate that the kidneys are the primary site of ghrelin clearance, which is significantly diminished in late sepsis. In addition, the liver also plays a role in the clearance of ghrelin, which was also reduced in late sepsis. The decreased clearance of ghrelin by the kidneys and liver may be due to renal and hepatic dysfunctions under such conditions. PMID- 14533010 TI - Identification and expression analysis of a novel splice variant of human Sprouty1 gene. AB - Sprouty (SPRY) was first identified in a genetic screen in Drosophila to be an antagonist of fibroblast growth factor (FGF) and epidermal growth factor (EGF) signaling, seemingly by inhibiting the Ras/MAP kinase pathway. During large scale DNA sequencing of the human fetal brain cDNA library, we cloned a novel splice variant of human Sprouty1 gene, and termed it human Sprouty1b (SPRY1b). It has the same deduced protein as Sprouty1a, which has been reported. And like other members of Sprouty family, SPRY1b deduced protein also has a C-terminal cysteine rich region. According to the search against human genome database, SPRY1b was mapped to 4q25-28. Expression analysis of SPRY1a and SPRY1b shows that they are hardly expressed in adult human, but have different expression patterns in fetus, which confirmed that SPRY1 is an important gene during fetal development. PMID- 14533011 TI - Experimental therapy of HPV16 induced tumors with IL12 expressed by recombinant vaccinia virus in mice. AB - Recombinant vaccinia viruses derived from strain Praha, clone P13, and strain MVA were used for intratumoral delivery and expression of IL12 genes in tumors induced by HPV16 E6+E7 oncogenes in mice. Intratumoral injection of 10(3) PFU of P13-IL12 virus resulted in an increase of intra-tumoral IL12 on days 6-13, while only low levels of IL12 were found in sera. After the inoculation of 10(6) PFU of MVA-IL12, the same levels of IL12 were found as in animals injected with control virus. The intratumoral inoculation of 10(3) PFU P13-IL12 resulted in only approximately 30% of the tumors being virus positive, which was a consequence of reduced multiplication of the recombinant virus in vivo. The number of virus positive tumors was not increased by repeated inoculations on three consecutive days. Intratumoral therapy with a dose of 10(3) PFU of P13-IL12 slowed down the growth of TC1 tumors, but never caused their regression. When local P13-IL12 treatment was combined with antigen-specific, DNA-vaccination therapy, no synergy between the two treatments was observed. The treatment with IL12-expressing virus retarded tumor growth to some degree, but did not change the number of regressing tumors. The highest efficacy of intra-tumoral P13-IL12 therapy was observed when the TC-1/A9 cell subline, with downregulated MHC class I expression, was used. TC 1/A9 tumors are less refractory to treatment with 10(3) P13-IL12/EL than are parental TC-1 cells. PMID- 14533012 TI - Human soluble tumor-associated antigens promote the suppression of rat mammary tumors by 5-fluorouracil and stimulate the functional activity of immune organs: Experimental and morphological studies. AB - This study examined whether the soluble 66 and 51 kDa tumor-associated antigens (sTAA) could promote suppression by the anticancer drug 5-fluorouracil (5-Fu) of chemically induced mammary tumorigenesis, and which, if any, morphological changes in the immune organs accompany this treatment. Dimethylbenzanthracene (DMBA, 8 mg/rat, twice) was used to induce mammary tumors. After the appearance of many large tumors, the preparations of sTAA and 5-Fu, alone or in combination, were administered in weekly doses, for 4 weeks. The following groups of mammary tumor-bearing rats were studied: 1) control non treated rats, 2) rats treated with sTAA, 3) rats treated with 5-Fu, 4) rats treated with 5-Fu and sTAA. The experiment was terminated when tumors in 70% of control rats became ulcerous. Treatment with sTAA alone significantly decreased tumor yield and their total area relative to controls. Both of these parameters showed an even larger significant decrease after treatment with 5-Fu, and the most marked decrease was obtained after the combined treatment with 5-Fu and sTAA. Results demonstrated that not only do sTAA have tumor-suppressive properties, they also enhance the anticancer effects of 5-Fu and prevent its toxic side effects. Morphologically, the treatment with sTAA was manifested in a significant increase in the size of the spleen follicles and mantle layer compared to control rats with large tumors. The treatment with 5-Fu decreased the sizes of almost all areas of the spleen compared to control rats, whereas the combined treatment with 5-Fu and sTAA increased all these parameters to the levels found in rats treated with sTAA alone. The total areas of the cortex and paracortex in the lymph nodes increased after treatment with sTAA. Treatment with 5-Fu alone resulted in a significant decrease of these areas which, as seen in the spleen, increased after combined treatment with 5-Fu and sTAA. Similar changes were seen in the areas of the separate lymph node zones. We concluded that the addition of sTAA to conventional tumor chemotherapy regimens has a remarkable synergistic effect on mammary tumors leading to curative antitumor responses of the host's immune organs. PMID- 14533013 TI - Differential effects of estradiol and raloxifene on collagen biosynthesis in cultured human skin fibroblasts. AB - The dose-dependent effect of a 24 h treatment with estradiol (E(2)) (1, 2, 5, 10 nM) and raloxifene (Rx) (1, 5, 10, 20 microM) on ER alpha and ER beta mRNA expression, collagen bio-synthesis, prolidase activity, MMP-2, MMP-9, insulin like growth factor I receptor expression (IGF-1R) and beta1-integrin expressions in cultured fibroblasts obtained from postmenopausal women were examined. Both ligands increased mRNA expression of ER compared to control. Rx at 5 and 10 microM concentrations had greater stimulative effect on collagen biosynthesis, prolidase activity and IGF-1R expression compared to E(2) at 2 and 5 nM concentration. Both studied ER ligands had no effect on beta1-integrin receptor expressions. MMP-2 expression was not detected in human skin fibroblast culture. In contrast to estradiol raloxifene inhibited the expression of MMP-9. Raloxifene had stronger positive stimulative effects on collagen biosynthesis, through different biochemical mechanisms, than estradiol in human skin fibroblasts and might reverse some of the postmenopausal changes in skin or connective tissue. Increase of collagen synthesis induced by raloxifene may be activated by both estrogen receptor dependent and independent pathways such as up-regulation of estrogen receptors, up-regulation of IGF receptor, transcriptional regulation of collagen genes by estrogen receptor-raloxifene complex, increasing of prolidase activity or finally by inhibition of MMP-9 expression. PMID- 14533014 TI - WNT2 and human gastrointestinal cancer (review). AB - WNT2 gene on human chromosome 7q31 is a paralog of the WNT2B gene on human chromosome 1p13. Rat Wnt2 gene was identified within rat genome draft sequence AC095247.4. Human WNT2 showed 96.4% total-amino-acid identity with rat Wnt2, 96.1% with mouse Wnt2, 68.6% with zebrafish wnt2, and 67.8% with fugu wnt2. WNT2 is an evolutionarily conserved secreted-type glycoprotein belonging to the WNT family. WNT2 mRNA is expressed in human fetal lung and placenta, but almost undetectable in normal gastrointestinal tract. WNT2 mRNA is frequently up regulated in human gastric cancer due to tumor-stromal interaction, and WNT2 gene is rarely amplified in human gastric cancer. WNT2 mRNA is also frequently up regulated in colorectal polyps, primary colorectal cancer of stage A-C, and also in liver metastasis from colorectal cancer. Putative biding sites for estrogen receptor, GATA-1, AP-2, TCF-1, BHLH, MBF-I, p53, and HNF-5 are located within the 5'-flanking region of human WNT2 gene. WNT2 is up-regulated by beta-estradiol in human MCF-7 cells; however, the mechanism of WNT2 up-regulation in most cases of gastrointestinal cancer remains to be elucidated. WNT2 is a tumor marker of gastric and colorectal cancer. Detection of theWNT2 protein in feces by immunohistochemistry or ELISA and WNT2 mRNA in feces by cDNA-PCR or custom-made microarray could be applied for screening of colorectal cancer. Because WNT2 up regulation leads to carcinogenesis through activation of the WNT-beta-catenin pathway, WNT2 specific antagonist might be applied for chemoprevention or treatment of gastrointestinal cancer. WNT2 gene is one of the targets for pharmacogenomics in the field of oncology. PMID- 14533015 TI - Identification and characterization of TRIP8 gene in silico. AB - TRIP1-TRIP15 genes encode thyroid hormone receptor beta (TR beta)-binding proteins. TRIP10 gene encodes FNBP1 family protein with FCH, FBH, and SH3 domains. Among 15 TRIP genes, TRIP8 gene remained uncharacterized except TRIP8 partial cDNA (L40411). Here, we determined the complete coding sequence of TRIP8 gene by using bio-informatics. Nucleotide sequence of full-length TRIP8 cDNA was determined in silico by assembling nucleotide sequences of FLJ14374 and DKFZp761F0118 cDNAs. TRIP8 protein (2540 aa) was found to consist of two bipartite nuclear localization signals (codon 352-368 and 2365-2381), TRI8H1 domain (codon 1697-1873), TRI8H2 domain (codon 2057-2351), and JMJC domain (codon 2387-2486). TRI8H1, TRI8H2 and JMJC domains were conserved among TRIP8, 5qNCA (C5orf7) and TSGA proteins. TR beta-binding domain was overlapped with N-terminal part of TRI8H2 domain, and C2HC4-type zinc finger-like motif was located within C terminal part of TRI8H1 domain. Because JMJC domain proteins are implicated in chromatin remodeling, TRIP8 was predicted to be a transcriptional regulator associated with nuclear hormone receptors. Human TRIP8 gene, consisting of 26 exons, was about 300 kb in size. Intra-species comparative genomics revealed that TRIP8-EGR2 locus at human chromosome 10q21.3 and 5qNCA-EGR1 locus at human chromosome 5q31 are paralogous regions within human genome. Microsatellite marker D10S1225, associated with Alzheimer's disease, non-syndromic congenital retinal non-attachment (NCRNA) and non-syndromic autosomal recessive persistent hyperplastic primary vitreous (arPHPV), was located within the TRIP8-EGR2 locus. This is the first report on comprehensive characterization of the TRIP8 gene. PMID- 14533016 TI - The contributions of John Money: a personal view. AB - John Money has been a dominant voice in sexology in the last part of the 20th century, breaking new ground in a wide variety of areas. In the process, he has been cantankerous, outspoken, and ever willing to do battle, but also original and thought provoking. This paper begins with an examination of science in general, moves on to psychology and sexology, and then examines Money s contributions to sexology in some detail. The latter are many and varied, including the development of the concept of gender, his theory of gender identity based on his work with intersex individuals, the John-Joan case, and his importance in establishing transsexualism as a diagnostic category and an academic discipline. Also important are his contributions to the development of the nomenclature of sexology, his importance to the sexology movement as a teacher, his significant research on a large variety of sexual topics, his ability to convince government agencies that sex was deserving of funding, and his association with the Erickson Educational Foundation. He also was a significant figure in the development of the Society for the Scientific Study of Sexuality (SSSS) and in raising the standard of its journal (The Journal of Sex Research), and therefore it is only fitting that an award be named after him. Though Money remains controversial, he has contributed significantly to the development of sexology as a discipline. PMID- 14533017 TI - History, causality, and sexology. AB - In 1896, Krafft-Ebing published Psychopathia Sexualis. Popularly defined as hereditary weakness or taintedness in the family pedigree, degeneracy was called upon as a causal explanation for perversions of the sexual instinct. Although Krafft-Ebing accepted Karl Ulrichs proposal that homosexuality could be innate and probably located in the brain, he paid little attention to neuropathological sexology. Alfred Binet challenged Krafft-Ebing's orthodoxy by explaining fetishism in terms of associative learning, to which Krafft-Ebing's response was that only those with a hereditary taint would be vulnerable. Thus did the venerable nature-nurture antithesis maintain its rhetoric, even to the present day. Krafft-Ebing died too soon to meet the Freudian challenge of endopsychic determinism, and too soon also to encounter the idea of a developmental multivariate outcome of what I have termed the lovemap. Like other brain maps, for example the languagemap, the lovemap requires an intact human brain in which to develop. The personalized content of the lovemap has access to the brain by way of the special senses. PMID- 14533018 TI - The effect of stimulus content on volunteering for sexual interest research among college students. AB - Much of the literature on volunteer bias in sexual interest research has been limited to either identifying variables on which volunteers differ from nonvolunteers or examining how the intrusiveness of the measurement device affects volunteer rates. The present study was designed to examine whether stimulus content would affect volunteer rates and volunteer/nonvolunteer differences in a large sample of college students (206 men and 358 women). The study also sought to determine whether such findings could also apply to research that recruits from exclusively heterosexual samples. Students completed questionnaires in small groups and were asked whether they would be willing to volunteer for studies that would involve viewing and rating five different types of sexually explicit images (nude men, nude women, heterosexual behavior, male homosexual behavior, and female homosexual behavior). Results indicate that men and women differed in the types of images that they would volunteer to view, based upon the content of the images. Furthermore, volunteers for each type of image reported significantly greater self-monitoring as well as sexual and general sensation seeking than did nonvolunteers, while differences on other measures were less consistent. Discussion is given to specific ways in which the findings and generalizability of sexual interest study results can be affected. PMID- 14533019 TI - Straw men and fairy tales: Evaluating reactions to A Natural History of Rape. AB - In this paper we respond to two frequent criticisms of our book, A Natural History of Rape (Thornhill & Palmer, 2000). The first criticism portrays the book as little more than a "just-so story" that human rape is an adaptation. We demonstrate that this portrayal is not accurate. The second criticism reflects a common response to the book s challenge of the popular assertion that rapists are not motivated by sexual desire but instead commit these crimes motivated by the urge to power, domination, and violence, and the urge to degrade and humiliate women. We demonstrate that such criticisms of our book are inherently contradictory and illogical. We believe it is important for sex researchers to understand that these sorts of criticisms are seriously flawed so that future research efforts toward understanding the causes of sexual coercion are not stalled. PMID- 14533020 TI - The amount and timing of parent-adolescent sexual communication as predictors of late adolescent sexual risk-taking behaviors. AB - The present study examined the moderating role of timing of first discussion of sexual intercourse with mothers and fathers on the relationship between the amount of sexual communication and sexual risk-taking behaviors in late adolescence. Late adolescents (N = 214) completed questionnaire measures regarding the amount and timing of sexual communication with their mothers and fathers. In addition, the participants provided information about their various sexual risk-taking behaviors. In general, the results suggest that timing of first discussion of sexual intercourse contributes additional variance in several sexually risky behaviors beyond that contributed by the amount of communication with both fathers and mothers. The implications of these findings are discussed. PMID- 14533021 TI - Sexual functioning and practices in a multi-ethnic study of midlife women: baseline results from SWAN. AB - This study examined the sexual practices and function of midlife women by ethnicity (African American, Caucasian, Chinese, Hispanic, Japanese) and menopausal status. Sexual behavior was compared in 3,262 women in the baseline cohort of SWAN. Participants were 42 to 52 years old, premenopausal or early perimenopausal, and not hysterectomized or using hormones. Analysis used multivariate proportional odds regression. In our sample, 79% had engaged in sex with a partner in the last 6 months, and a third considered sex to be very important. Common reasons for no sex (n = 676) were lack of partner (67%), lack of interest (33%), and fatigue (16%). Compared with Caucasians, Japanese and Chinese women were less likely, and African Americans more likely, to report sex as very important (p < 0.005). Significant ethnic differences were found for frequency of all practices. Perimenopause status was associated only with higher frequencies of masturbation and pain during intercourse. PMID- 14533022 TI - Puerto Rican drug users experiences of physical and sexual abuse: comparisons based on sexual identities. AB - This study integrates the results of quantitative and qualitative methods to elucidate the association between sexual identity and physical and sexual abuse among Puerto Rican drug users. A structured questionnaire was administered to 800 subjects in New York and 399 in Puerto Rico. A total of 93 subjects (7.9%) self identified as homosexual or bisexual. Gay males were significantly more likely than heterosexual males to report first occurrence of physical abuse by a family member in childhood. Both gay and bisexual males were more likely than their heterosexual counterparts to report first experiencing unwanted sex in childhood and intimate partner physical abuse later in life. Lesbians were more likely than female heterosexuals to report unwanted sex in childhood. Qualitative data were collected through in-depth life histories with 21 subjects and suggest that gay and lesbian subjects perceive antihomosexual prejudice on the part of family members as one cause of childhood physical and sexual abuse. PMID- 14533023 TI - The construction and psychometric properties of an instrument measuring support for sexual rights. AB - The purpose of this paper is to report the findings of an attempt to construct and examine the psychometric properties of an instrument that measures support for the 11 sexual rights formulated by the World Association of Sexology. Since international conferences held in the mid-1990s, a new approach to sexual issues has been promoted by health professionals, sex educators, and human rights advocates. This approach emphasizes the empowerment of individuals and examines sexual health concerns within a human rights framework. Goals of a sexual rights approach have been to promote awareness and advocacy of sexual issues and especially to facilitate the work toward better sex education and sexual health services. Analysis of reliability and validity data from two university samples (n = 388 and n = 175) indicated that the majority of the sexual rights scales have acceptable psychometric properties. This paper concludes with a discussion of ways to improve the scales for future use and the need to validate the scales using samples that allow generalizability of findings. PMID- 14533024 TI - Ambivalent sexism and attitudes toward women who engage in premarital sex in Turkey. AB - This study examined the relationship of ambivalent sexism, political conservatism, demographic variables (age, education, and gender), and prior sexual experience to Turkish men s and women s attitudes toward women who engage in premarital sex. Participants included 124 Turkish undergraduate students and 60 nonstudent Turkish adults. Benevolent but not hostile sexism uniquely predicted more negative views of women who engage in premarital sex once other variables were controlled. Regression analyses demonstrated that for both men and women, older, more politically conservative and less sexually experienced respondents and more educated men (but not women) respondents were more likely to disapprove of women who engage in premarital sex. Similarly, regression analysis revealed that men who were older, politically conservative, and less sexually experienced expressed stronger preferences for marrying a virgin. Both hostile and benevolent sexism predicted men s preference for marrying a virgin after all other variables were controlled. PMID- 14533025 TI - The myth of innocence: sexual scripts and the recognition of child sexual abuse by female perpetrators. AB - Although child sexual abuse has been studied extensively, minimal attention has been paid to sexual abuse by females. This paper explores the prevalence of female sex offending and reveals the paradoxes that exist within the available data. Moreover, it highlights the role of traditional sexual scripts in impeding the official recognition of the problem. Traditional sexual scripts, particularly the perception of females as sexually passive, harmless, and innocent, appear not only to have influenced broader societal views concerning sexuality and sexual abuse but also to have permeated the criminal law, victim reporting practices, and professional responses to female sex offending. The implicit denial of women s potential for sexual aggression within these three domains may ultimately contribute to the underrecognition of the problem in official sources. PMID- 14533026 TI - Methotrexate in inflammatory eye disease. PMID- 14533027 TI - Chemokines--their role in immunotherapy for intraocular inflammation. AB - Chemotactic cytokines are responsible for leukocyte migration and the immunopathogenesis of various inflammatory lesions. Together with other types of cytokines, chemokines play a major role in inducing/regulating inflammation and various immune responses. By targeting chemokines, immunotherapies could become another option for treating patients with uveitis. Indeed, a variety of chemokine based therapies have been tested for their possible application for various pathological diseases, including intraocular inflammation. An example of chemokine-based therapy is anti-tumor necrosis factor (TNF)-alpha therapy, a very successful treatment. Chemokine- and cytokine-based therapies, therefore, appear to be a promising choice for the treatment of intraocular inflammation. PMID- 14533028 TI - Participation of pigment epithelium in ocular immune privilege. 3. Epithelia cultured from iris, ciliary body, and retina suppress T-cell activation by partially non-overlapping mechanisms. AB - PURPOSE: The ocular microenvironment is immunosuppressive and anti-inflammatory. Since various ocular pigmented epithelia contribute to this microenvironment, we studied the relative capacities of pigment epithelial (PE) cells cultured from the iris, ciliary body, and retina of mouse eyes to suppress T-cell activation in vitro. METHODS: Pigment epithelium was cultured from iris, ciliary body, and retina for 14 days, then assayed for the capacity, directly or across transwell membranes, to suppress mixed lymphocyte reactions and anti-CD3 stimulation of T cells. Potential molecules responsible for suppression were examined by attempting to block suppression with appropriate reagents, and by using mice with pertinent mutant or disrupted genes. RESULTS: We found that PE cells from all three ocular tissue sources profoundly suppressed T-cell activation in vitro. While iris PE suppressed poorly when separated from T cells by a transwell membrane (implying that cell contact is necessary), retina PE suppressed fully even in the presence of such a membrane (implying that soluble factors were responsible). Ciliary body PE used both soluble factors as well as cell contact to achieve suppression. Suppression could not be ascribed to TGFbeta, IFNgamma, TNFalpha, CD48, or ICAM-1, or to interactions between CD40 and CD154, or CD95 and CD95 ligand. Galectin-1, a galactoside-binding protein, was found to be expressed on all cultured PE cells, but only retinal pigment epithelium (RPE) from galectin 1 KO mice showed reduced capacity to inhibit T-cell activation. CONCLUSIONS: Cultured pigment epithelia from iris, ciliary body, and retina comparably suppress T-cell activation in vitro, but by partially different mechanisms. Although RPE cells suppress in part through expression of galectin-1, the molecular mediators of suppression by iris and ciliary body PE remain to be identified. PMID- 14533029 TI - Prognostic significance of Fas expression in retinoblastoma. AB - PURPOSE: The loss of Fas is believed to enhance metastatic spread in diverse human tumors. We studied the expression of Fas by immunohistochemistry in retinoblastomas and correlated the results clinicopathologically. METHODS: Archival specimens of 76 retinoblastomas were included in this study. The tumors were divided into three groups: Group A (n = 21) comprising tumors with no invasion; Group B (n = 46) tumors with invasion of choroid (diffuse), optic nerve (post-laminar), and orbit; and Group C (n = 9) tumors with both invasion and metastasis. Sections were immunostained with monoclonal antibody CD95 (APO-1/Fas) and the immunoreactivity was assessed. RESULTS: Negative immunoreactivity of Fas was observed in Group B and Group C tumors (p < 0.001). Poorly differentiated retinoblastomas showed a decreased Fas expression (p = 0.006). CONCLUSIONS: Decreased expression of Fas is seen in aggressive tumors and tumors that are poorly differentiated. This preliminary observation deserves further investigation, which may then shed more light on the immune escape mechanisms of this tumor and thus enable novel therapeutic strategies. PMID- 14533030 TI - The roles of T-lymphocyte subpopulations (CD4 and CD8), intercellular adhesion molecule-1 (ICAM-1), HLA-DR receptor, and mast cells in etiopathogenesis of pterygium. AB - PURPOSE: The etiopathogenesis of pterygia is controversial. We wished to explore the effects of inflammatory cells and mediators such as T-lymphocyte subpopulations (CD4 and CD8), mast cells, intercellular adhesion molecule-1 (ICAM 1), and HLA-DR receptors on pterygium development. METHODS: Immunohistochemical stainings were performed for T-lymphocyte subpopulations, mast cells, ICAM-1, and HLA-DR on sections of pterygium specimens. Nasal bulbar conjunctival specimens of patients undergoing cataract or vitrectomy operations were used for comparision. The mean number of mast cells/mm2 was calculated using an ocular grid, and the difference between the numbers derived from pterygium and from normal conjunctival sections was analyzed statistically using the Mann-Whitney U test. CD4 and CD8 lymphocytes in the epithelial and subepithelial regions, and ICAM-1 and HLA-DR expression on pterygium epithelium were compared qualitatively. RESULTS: Increased CD4 and CD8 lymphocytic infiltration in pterygium sections was observed. Only occasional lymphocytes were detected in normal conjunctival sections. The CD4/CD8 ratio was 0.33 for pterygium epithelium and 1.34 for pterygium substantia propria. Increased HLA-DR and scattered ICAM-1 expression were also detected on pterygium epithelium, but not on the normal conjunctival epithelium. The difference in mast cell numbers between the pterygium and control groups was not statistically significant. CONCLUSIONS: The detection of T lymphocyte infiltration and ICAM-1 and HLA-DR expression in pterygium epithelium strongly supports the suggestion that cellular immunity plays an important role in pterygium formation. Pterygium epithelium also seems to participate actively in the augmentation of this inflammatory process by expressing ICAM-1 and HLA-DR molecules. Mast cells may participate in some stages or subtypes of pterygium during its development. PMID- 14533031 TI - The relationship between soluble L-selectin and the development of diabetic retinopathy. AB - Diabetic retinopathy is a leading cause of adult vision loss and blindness. Earlier studies have shown that polymorphonuclear neutrophils play an important role in the pathogenesis of diabetic vascular complications. Stimulation of these cells is associated with the desquamation of L-selectin. The aim of the study was to evaluate the relationship between the serum concentration of L-selectin and the development of retinopathy in Type 2 diabetic patients. The study comprised 51 Type 2 diabetic patients, aged 65.2 +/- 7.5 years, with a diabetes duration of 10.1 +/- 8.9 years and HbA1c of 8.2 +/- 1.4. The study subjects were divided into two groups: Group A diabetic patients with retinopathy (n = 28) and Group B diabetic patients without retinopathy (n = 23). Twenty age- and sex-matched nondiabetic healthy subjects were enrolled as the control group. Patients with any inflammatory disease were excluded. Retinopathy was assessed by centrally graded retinal photographs. The serum concentration of sL-selectin was estimated using an ELISA test. We observed significantly higher serum concentrations of sL selectin in Type 2 diabetic patients with retinopathy than in healthy subjects (36.5 +/- 18.1 vs. 11.4 +/- 7.5 ng/ml, p < 0.001). There was also a significant difference between Group A and Group B (36.5 +/- 18.1 vs. 24.2 +/- 13.5 ng/ml, p < 0.05) as well as between Group B and the controls (24.2 +/- 13.5 vs. 11.4 +/- 7.5 ng/ml, p < 0.01) with regard to sL-selectin levels. sL-selectin was significantly correlated with HbA1c (r = 0.93, p < 0.001) and with diabetes duration (r = 0.44, p < 0.001). These results suggest that there was a strong relationship between sL-selectin and diabetic retinopathy. The strong correlation between sL-selectin and HbA1c levels supports the concept that the sL-selectin level is increased with poor glycemic control, which may affect endothelial cell activity and cause subsequent microvascular complications. PMID- 14533032 TI - Methotrexate as a first-line corticosteroid-sparing therapy in a cohort of uveitis and scleritis. AB - PURPOSE: To evaluate the clinical experience with methotrexate as a first-line corticosteroid-sparing drug in patients with resistant ocular inflammation. METHODS: We retrospectively studied 39 consecutive patients with uveitis (n = 36) or scleritis (n = 3) who were treated with methotrexate following inadequate control with corticosteroids lasting five years. Criteria for initiating treatment with methotrexate and defining outcome were strictly defined. RESULTS: The cohort included 21 females and 18 males, all Caucasians, with a mean age of 26.6 years (range: 3-73 years). Patients were followed up for 21.5 +/- 12.6 months. Treatment was discontinued due to side effects in 10 patients (26%). Of the remaining 29 patients, full or partial control of inflammation was achieved in 23 (79%). Response to treatment was observed after a mean of 2.4 +/- 0.8 months. Ten patients were fully controlled and discontinued methotrexate therapy after a mean of 20.9 +/- 9.2 months, with no recurrence of inflammation. Use of topical and systemic corticosteroids was markedly reduced in responsive patients. CONCLUSIONS: Methotrexate is recommended as a first-line adjunct to or replacement of systemic corticosteroids in the treatment of ocular inflammation. PMID- 14533033 TI - Successful treatment with combination of systemic antiviral drugs and intravitreal ganciclovir injections in the management of severe necrotizing herpetic retinitis. AB - PURPOSE: To report the use of intravenous (IV) antiviral agents and intravitreal ganciclovir injections in three immunocompetent patients with severe acute retinal necrosis (ARN). METHOD: Case series. RESULTS: Three immunocompetent patients, who had lost vision in the first eye due to ARN, received intensive treatment with IV foscarnet or acyclovir or ganciclovir and intravitreal ganciclovir injections for the treatment of severe ARN involving the fellow eye. The retinitis resolved and final visual acuity of the fellow eye improved to 20/20 in all three cases after a mean follow-up of 17 months. CONCLUSIONS: Intensive treatment with a combination of two intravenous antiviral drugs and intravitreal ganciclovir injections was successful in the management of patients with acyclovir-resistant ARN and monocular status. PMID- 14533034 TI - Ocular filariasis due to Wuchereria bancrofti presenting as panuveitis: a case report. AB - PURPOSE: To report a case of ocular filariasis due to Wuchereria bancrofti manifesting as panuveitis. DESIGN/METHODS: Observational case report. RESULT: A 30-year-old, healthy, Asian, Indian male presented with decreased vision and panuveitis with secondary glaucoma in the right eye. Careful slit-lamp examination revealed multiple, tiny, motile larvae in the anterior chamber. Indirect ophthalmoscopy showed vitritis with plenty of vitreous membranes, and subretinal yellow lesions in the peripheral retina along with retinal pigment epithelial tracts. An aqueous tap and a peripheral blood smear isolated microfilariae of W. bancrofti. Therapy with diethyl carbamazine citrate along with systemic steroids provided symptomatic relief. CONCLUSIONS: Ocular filariasis due to W. bancrofti is very rare. It can present in an otherwise asymptomatic patient without any constitutional symptoms. Careful examination, prompt diagnosis, and early treatment can reduce ocular morbidity. PMID- 14533035 TI - Severe allergic blepharoconjunctivitis induced by a dye for eyelashes and eyebrows. AB - PURPOSE: To report a case of severe allergic blepharoconjunctivitis induced by a dye for eyebrows and eyelashes. METHODS: A 38-year-old healthy female was examined one day after dyeing her eyebrows and eyelashes with a black cream dye intended for this purpose (FDA-approved). The patient complained of severe eye itching, redness, and epiphora. RESULTS: Vision was 20/40 in both eyes; a moderate edema of the eyebrows and eyelid margins was noted. The conjunctiva was severely hyperemic with papillary reaction and chemosis. The corneas, anterior chambers, irides, lenses, and posterior segments were normal. The patient was treated with Dexamethasone 0.1% and ocular lubrication. After five days of treatment, the ocular symptoms improved, her vision returned to 20/20 in both eyes, and the ocular examination was within normal limits. CONCLUSION: FDA approved dyes for eyebrows and eyelashes can sometimes irritate the ocular surface. PMID- 14533036 TI - [Outpatient primary stent-angioplasty in symptomatic internal carotid artery stenoses]. AB - In 81 patients, average age 69.3 (57-79) years with symptomatic ICA stenosis stent implantation was performed under outpatient conditions. An interdisciplinary consensus ascertained that the patients were suitable for outpatient therapy. In accordance with the NASCET criteria all patients belonged to the high-risk group. Within 120 days before the intervention, the following clinical events had occurred: 32 minor strokes, 43 TIA, 20 amaurosis fugax. 61/81 patients revealed a bulbar, 20/81 a postbulbar ICA stenosis, the average degree of stenosis was 88.6 % (75-97). Stent application, using 3 different stent models, and subsequent PTA were always performed under protection. The transfemoral route was always treated with an endovascular closure system. All patients were given follow-up examinations by means of colour duplex sonography (CDS) of the neck vessels on days 1, 30, 180 and 360, and contrast-medium intensified MRT on day 5 post-intervention. The technical success rate with regard to stenting and PTA was 100 %. One case of hypotonic dysregulation occurred during the intervention. Atheromatous particles and plaque fragments were found in the protection system in 3/81 (3.7%) of the patients. A plaque embolism was excluded clinically and in the contrast-medium intensified MRT 5 days after the intervention in 79/81 (97.5%) of the patients, while two patients were found to sustain an asymptomatic cortical barrier impairment. After 180 days, no restenosis or stent dislocation were found in the CDS, all ECAs were perfused. After 360 days, one asymptomatic occlusion and four restenoses > 70% were found (5/81 = 6.2% restenosis rate after 1 year), which were successfully treated by angioplasty. The method presented bears a low complication rate and shows a high success rate. If patient recruitment is adequate, this intervention can safely be performed on an outpatient basis. PMID- 14533037 TI - [Infrainguinal bypass grafting in patients with end-stage renal disease and critical limb ischaemia: is it worthwhile?]. AB - Patients with end-stage renal disease (ESRD) constitute an increasing proportion of patients undergoing infrainguinal bypass surgery for critical limb ischaemia (CLI). The aim of this retrospective study was to determine graft patency, healing of pedal lesions, limb salvage and survival following infrainguinal arterial reconstruction in this high-risk subset of patients. 34 patients with ESRD undergoing 37 bypass procedures for CLI (rest pain 2; tissue loss 35) were identified from the vascular registry. These included 13 femoropopliteal and 24 femorotibial bypasses with autogenous (67.6%) or prosthetic (32.4%) materials. The median age in this series was 62 years and 79% were diabetics. Using life table analysis, the cumulative primary patency rate was 88% at 1 month and 81% at 2 years. The resulting limb salvage rate amounted to 94 and 86% at 1 month and 2 years, respectively. Healing of the pedal lesions was accomplished in only 50% of patients at 6 months. Toe lesions could be treated more successfully than forefoot and deep heel defects (p = 0.04). With a perioperative mortality of 3/37 cumulative survival rate declined to 21% at 2 years. Late mortality correlated significantly with a history of previous myocardial infarction or congestive heart failure (p = 0.001). Infrainguinal revascularisation can be performed in dialysis-dependent patients with acceptable patency and limb salvage rates. However, bypass grafting should be mainly reserved to patients without severe cardiac disease and to those without extensive tissue loss. PMID- 14533038 TI - [Limb salvage in diabetic foot syndrome with pedal bypass using the in-situ technique]. AB - INTRODUCTION: Treatment of the ischemic diabetic foot syndrome still represents a medical and economic challenge. Contrary to the aims of the Saint Vincent declaration a dramatic reduction of major amputations in Germany was not noted, although in the diabetic patients the predominant type of tibial artery occlusion allows construction of pedal bypasses for limb salvage. METHOD: In patients with ischemic diabetic foot syndrome following angiographic evaluation of the ischemic limb, the indication for surgical revascularisation of patent pedal arteries was established. The in-situ technique was preferred in the presence of a suitable ipsilateral greater saphenous vein whenever possible. Revascularisation was followed by treatment of foot ulcerations or, if necessary, minor amputations. Patients were followed by clinical examination and duplex scan investigation of the bypass in regular intervals. RESULTS: From 01/89 to 12/01 in 79 patients (59 men and 20 women) with non healing ulcerations or established gangrene from a total of 175 pedal bypasses 84 pedal bypass operations in 84 limbs were performed using the in-situ technique. All patients were diabetic and in addition 13.9% were dependent on hemodialysis for end stage renal disease. 59.5% of the bypasses originated from the popliteal artery (distal origin bypass). The dorsalis pedis artery was chosen for the distal anastomosis in 83% and the posterior tibial artery in 17%. Two patients (2.4%) died postoperatively from cardiac events. Early bypass occlusion occurred in 8.4% resulting in a major amputation rate of 6%. After 60 months primary, primary assisted and secondary patency was 67.7%, 71.5% and 75.3% respectively with a limb salvage rate of 78%. CONCLUSION: Pedal bypass using the in-situ technique provides excellent long term limb salvage rates in a disease with a generally unjustified bad prognosis with respect to limb salvage. PMID- 14533039 TI - [Short distal origin vein graft in diabetic foot syndrome]. AB - PURPOSE: World wide increase of diabetes compound with diabetic foot syndrome becomes a challenge in vascular surgery to avoid limb loss. In diabetics a special pattern of atherosclerosis is prevalent with disease limited to the infrageniculate arteries but sparing inflow vessels and distal tibial and pedal arteries. This provides short bypass grafting from popliteal to tibial and pedal arteries, a concept first described by F. Veith in 1981. METHODS: Diabetics with severe atherosclerotic disease and limb-threatening ischemia got general evaluation and vascular imaging. Falling in this special category the patients underwent short vein bypass grafting originating at the first or third popliteal segment extending to the tibial or pedal arteries. Follow up of patency and limb salvage was nearly complete. RESULTS: From 1988 to 2001 124 diabetics received 140 vein bypass grafts for limb salvage, 95.7% already preoperatively with foot necrosis. Operative mortality rate was 1.4%, major morbidity rate was 9.3%, early graft failure rate 8.5% and early amputation rate was 3.8%. 2 year primary patency, primary assisted patency, secondary patency rates and limb salvage were 73.3%, 75.7%, 76.4% and 87.2%. 5 years results were 63.6%, 69.2%, 70.0% and 81.9% respectively. DISCUSSION: This series revealed exceptionally good results in patients with diabetes mellitus after short vein bypass grafting in concert with earlier studies since 1981. Compared to long femorodistal grafts there was no difference in longterm patency. Bypass grafting in diabetic foot syndrome is still regarded to have a poor prognosis. Just the contrary is the case. This study in concert with former studies revealed distal origin bypass grafting a durable and effective procedure to fight limb loss in diabetic foot syndrome. CONCLUSION: Distal origin vein bypass grafting is an excellent method just for patients with diabetes. Looking at limb salvage rates over 80 % in 5 years, this procedure should be offered more often suitable patients with diabetic foot syndrome. PMID- 14533040 TI - [Borderline amputations in diabetics--open questions and critical evaluation]. AB - Minor amputations are frequently performed in neuroischemic or neuropathic lesions of the diabetic foot. In German-speaking countries, the term "Grenzzonenamputation" was created to describe the combination of minor amputation exactly in the border zone to vital tissues, necrosectomy and debridement. For early and optimal rehabilitation, as much vital tissue as possible should be conserved, especially the skeletal structures of the foot. "Grenzzonenamputationen" are of utmost importance for the prevention of ascending infections, and they reduce the duration of clinical and outpatient treatment. "Grenzzonenamputationen" should be performed only by experienced surgeons and if the arterial perfusion is sufficient. They should be as tissue-sparing as possible, and structured interdisciplinary postoperative care is mandatory. Controversial opinions exist with respect to the use of tourniquets, conservation or resection of cartilage and sesamoids, open amputation or primary closure of the wound, interdigital spacer function of toes, aseptic proximal transection of tendons, postoperative wound care, and antibiotic therapy. In view of these controversies, the most important distal minor amputations are described and discussed, with special regard to the habitual surgical way of thinking. PMID- 14533041 TI - [Diabetic neuropathic osteoarthropathy (Charcot foot)]. AB - Diabetic neuropathic osteo-arthropathy (DNOAP; Charcot's foot) is a dramatic complication of diabetic polyneuropathy. The diabetic foot is the most neglected long-term sequela of diabetes mellitus, and this is especially true for DNOAP. The increasing number of diabetics presenting in the foot ambulance with DNOAP demonstrates this general lack of knowledge and indicates that this complication is more frequent than hitherto assumed. In view of this dilemma, the pathogenesis, diagnosis and therapy of DNOAP are reviewed. Special emphasis is given to the differentiation between the neuropathic and neuroischemic foot as well as between acute DNOAP and bacterial infection. Therapy of DNOAP is predominantly conservative. The indication for surgery should be restricted to serious deformities of the foot, instability of joints, imminent skin perforation caused by fracture-induced dislocation of bones, and recurrent ulcers caused by prominent bones. Surgery comprises ulcer excision, ablation of bony prominences, as well as orthopedic surgery for the reconstruction of the foot skeleton. PMID- 14533042 TI - [Hemobahn-endoprosthesis: long-term experience (< or = 4 years follow-up) with percutaneous application in stenoses and occlusions of the superficial femoral artery]. AB - We tested the value of hemobahn-endoprosthesis in a prospective, nonrandomized, clinical trial in PAD-patients with femoral artery occlusive disease. During 4 years we placed in 52 patients (55 extremities) a total of 78 hemobahn endoprostheses for treatment of stenoses (15%) or occlusions (85%) of femoral artery. The average length of the treated lesions was 11.6 cm, the mean length of the hemobahn-endoprosthesis was 15.9 cm, the average device diameter 6.1 mm. In 45% of the patients we used cross-over and in 55% ipsilateral antegrade technique. Fontaine's clinical stages were II b (89%), III (2%) and IV (9%); there were 15 diabetics. The average period of follow-up was 33 months (1-52 months). The primary/secondary patency rates after the first year (n = 50) were 64% vs. 78%, after 2 years (n = 46) 61% vs. 76%, after 3 years (n = 32) 53% vs. 66% and after 4 years (n = 14) 29% vs. 50%. After the first experiences in 1998 with 10 stent-occlusions in 14 treated extremities, indications were checked and thereafter hemobahn was only used in "ideal indications" as segments with a length of > or = 1 cm proximal and distal without any obstruction, no lesions in the popliteal artery, at least one open lower leg vessel and no severe calcifications. The primary/secondary patency rates were significantly better after exclusion of nonideal patients treated during the first year: Thus, primary/secondary patency rates were 76 % vs. 90% after first year (n = 32), 74 % vs. 89% after second year (n = 38) and 71% vs. 83% after third year (n = 24); after 4 years of 8 treated arteries 4 were primary and 7 were secondary open. Hemobahn-endoprostheses are suitable for treatment of long femoral artery occlusions. The medium patency rate in "ideal indications" is higher than that obtained by angioplasty with or without uncovered stents. PMID- 14533043 TI - [Knotted intravascular catheters--what should be done?]. AB - INTRODUCTION: Fixed intravasal catheters are mainly caused by knots. Removal can be achieved by intervention or surgical exploration, but this is associated with additional morbidity and mortality. METHODS: 2 patients were operated for knotted catheters in our institution during the last 2 years, and their records are demonstrated. Treatment options, possible complications, catheter types and locations of knotting are analyzed by a medline search. RESULTS: the search revealed the data from 115 patients. 53 (46.1%) of all "lost" catheters were Swan Ganz catheters. In 60.9% the catheters could be removed by radiological interventions. Open revision was necessary in 33% of all cases. The catheters were left in place when the clinical condition of the patient did not allow removal (n = 7). However, these patients suffered from a high mortality (5 of 7 patients). Over all mortality reached 8.7%. In the own two cases one removal by sternotomy and one by exploration of the right internal jugular vein were necessary, both operations succeeded without complications. CONCLUSION: Most of all "lost" intravasal catheters are removed by radiological intervention; only one third needs open surgical therapy. These procedures are harmful for the patient and bear considerable risks for complications. PMID- 14533044 TI - [Does venous thrombectomy still play a role in the treatment of Paget-von Schroetter-Syndrome?]. AB - In a retrospective study we reviewed 24 patients treated by venous thrombectomy for acute thrombosis of the axillary vein during the past 23 years. We performed 25 venous thrombectomies without any complications. Complete recanalisation could be achieved in all cases of acute thrombosis. Whereas similar clinical results can be achieved by thrombolytic therapy, complete recanalisation by thrombolytic therapy is only achieved in 60% of cases. PMID- 14533045 TI - [Dialock--an alternative to Permcath and access graft]. AB - PURPOSE: An adequate vascular access is the precondition for a well-functioning hemodialysis. Due to the increasing age and the rising co-morbidity of hemodialysis patients the number of those with grafts or central venous catheters (CVC) is steadily growing. The Dialock vascular access system provides a subcutaneously implantable device for hemodialysis that combines the advantages of central venous access with percutaneous puncture. PATIENTS AND METHODS: Over a period of 30 months 26 Dialock vascular access systems were implanted at our department. 17 patients were male, 9 patients female. In 11 patients the implantation of this system was indicated for internal medicine reasons. In 15 long-term hemodialysis patients implantation was carried out as ultimate solution because of persisting vascular access problems. In a comparable control group of 22 hemodialysis patients 47 vascular accesses were registered within the same period. In parallel, another 110 patients with grafts implanted at our department at that time but not put to hemodialysis at our clinic were analysed. RESULTS: In a comparable number of access days the graft and the central venous catheter showed a considerably higher infection rate (Exit site + blood stream infection) per 1,000 hemodialysis access days than the vascular access device. The rate of malfunctions with or without subsequent lysis was higher using the CVC than when applying Dialock or graft. Angiographies with percutaneous transluminal angioplasty (PTA), or thrombectomies, respectively were significantly more often required with the graft than changes of catheter when using the vascular access system. More than half of the 110 patients observed at the same time, who had a graft implanted at our department without subsequent hemodialysis at our clinic had to undergo another surgical or radiologic intervention. CONCLUSION: The Dialock access system represents in our experience a safe and with careful application less complicated vascular access device for the complex hemodialysis patient. With comparatively low infection and complication rates the Dialock vascular access system serves as a good alternative to the permanent CVC (Permcath) as well as to the graft. PMID- 14533046 TI - [The stenosed Brescia-Cimino fistula: operation or intervention?]. AB - INTRODUCTION: Stenosis of the vein close to the arteriovenous anastomosis is the most frequent cause for late failure of Brescia-Cimino fistulae (BCF). Although since decades proximal re-anastomosis has been regarded as the surgical standard treatment, success rates can hardly be deducted from the literature. Considering the increasing activities of interventional radiologists surgical position finding seems necessary. METHODS: Over three years 30 anastomotic BCF stenoses were treated in 28 patients. In 15 patients the stenosis had caused fistula thrombosis. In all cases the fistula vein was re-anastomosed to the proximal radial artery. All patients could be followed up (average 12 months). Each fistula functioning after 24 hours was classified as procedural success. For calculation of patency rates (life table analysis), however, usability of the needling segment of the access was assessed. RESULTS: Procedural success rate was 100%. One fistula thrombosed on the second postoperative day after a successful dialysis session due to an overlooked proximal stenosis of its feeding radial artery. In five fistulae stenoses developed after 4 to 13 months. In only one fistula this was a true re-stenosis of the newly created anastomosis (0.03 per patient-year). Two of the stenoses occurred in the needling segment of the access vein and within its central venous outflow, respectively. Overall re-intervention rate was 0.3 per patient-year, and primary (secondary) patency was 80% (95%) at one year and 67% (87%) at two years. CONCLUSION: In BCF proximal re-anastomosis is a simple and effective therapeutic option for anastomotic venous stenosis. Re intervention rates are low and procedural success rates as well as primary and secondary patency rates at least equal those of interventional radiology. PMID- 14533047 TI - [Impact of ultrasonographic vascular mapping on constructing autogenous arteriovenous fistulas for permanent hemodialysis access?]. AB - PURPOSE: We used high resolution ultrasonography to identify usable veins and arteries in the forearm for creation of autogenous arteriovenous fistulas (AVF) for permanent hemodialysis access. The effect of preoperative vascular mapping followed by intraoperative controls on the outcome of AVF should be reported. METHODS: study subjects were adults referred for primary permanent hemodialysis access between January 2001 and November 2002. In all patients sonographic assessment was performed before surgical evaluation. A feeding artery was considered adequate if the diameter was more than 1.5 mm, the vein more than 2.0 mm. All AVF were controlled by intraoperative sonographic measurements of PSV and diameter of the fistula-vein. RESULTS: AVF were placed in 94.1% of all patients. The early failure rate of AVF was 6.3%. Primary patency rate of AVF after 24 hours: 93.7%, after 30 days: 91.4%, after 3 months: 86.9%, preliminary patency rate after 1 year: 70.1%. No unsuccessful surgical explorations were performed. 85.5 % of AVF were constructed as forearm fistulas. Suboptimal vessels (artery < 2.0 mm, vein < 3.0 mm) were used in 31.3% of patients. Patency rates did not differ in this subgroup. In 2 patients synthetic grafts were placed because of non-maturation of AVF. In 15 patients the AVF had to be cannulated by experts for 3 to 6 months. 50.0% of all AVF were constructed in diabetic patients. Patency rates were equal to that of non-diabetic patients. No patient suffered on signs of steal-syndrome. CONCLUSION: the assessment of forearm vessels by high resolution sonographic vascular mapping helps to find the optimal location for constructing an arteriovenous wrist fistula in almost all patients needing a permanent hemodialysis access. The aggressive approach to the creation of autogenous fistulas could be realized without unsuccessful surgical explorations and with a minimal early failure rate, a high maturation rate including patients with diabetes mellitus and no signs of steal-syndrome PMID- 14533048 TI - [Iliac artery-ureteral fistula and its treatment]. AB - Arterio-ureteral fistulas are rare. A life-threatening complication is severe hemorrhage. More than 50 per cent of the cases were reported during the last ten years. The predisposing causes mainly are operations of malignomas of the kidney and the efferent urinary tract as well as gynecological neoplasias with or without irradiation. Stenting of the ureter or reconstructive vascular surgery may also cause arterio-ureteral fistulas. Because of the intermittent symptomatology diagnosis may be very difficult. Therapy and prognosis for life as well as for organ failure depend on pretherapeutic examination. In former times surgical repair was the therapy of choice. With the introduction of interventional techniques morbidity and mortality decreased significantly. During the last five years three patients with iliac-ureteral fistulas were treated in our department. In two cases the fistulas was excluded by covered stent grafts, in one case coiling alone of the internal iliac artery was successful. Reviewing the literature and our experience with arterio-ureteral fistula angiography is recommended for diagnostics. Covered iliac stent grafts are the therapeutic option of choice. The leakage of the ureter needs not to be repaired mandatoryly. PMID- 14533050 TI - Provision of speech-language services in the schools: working with the law. AB - Decisions regarding eligibility for speech-language services in the schools can be the most difficult decisions facing school-based speech-language pathologists. Clinicians and parents must be informed about the federal mandates for services in the schools for children with special needs in order to make appropriate decisions and to ensure that children with disabilities are receiving the services to which they are entitled. This article discusses the Individuals with Disabilities Education Act (IDEA), the federal law guaranteeing school-based services to children with disabilities, as an educational planning process that promotes meaningful access to the general curriculum for all students. Steps are then outlined for making eligibility decisions by following this established process and using IDEA's goals as the standard by which services are developed and implemented. Examples are given linking speech-language services to state learning standards. PMID- 14533051 TI - Speech-language caseloads in the schools: a workload analysis approach to setting caseload standards. AB - This article highlights key concepts of the recently revised policy documents of the American Speech-Language-Hearing Association on workload and caseload size for speech-language pathologists (SLPs) in schools. A major premise is that the total workload activities required and performed by school-based SLPs must be taken into account to set appropriate and reasonable caseload standards. Current policies and practices used to define caseloads appear to be inadequate to ensure caseloads that allow enough time for the provision of quality services. In this article we describe school-based SLPs' workload, including the expanded roles and responsibilities that are mandated by the Individuals with Disabilities Education Act (IDEA) and other legislation. We consider the available research on the effects of large caseloads on the service options chosen by SLPs, treatment outcomes for students, and SLP attrition and burnout. Finally, we present an approach for analyzing a SLP's total workload activities, along with a discussion of how that analysis can be used to help determine caseload standards consistent with meeting the intent of IDEA and the needs of students with disabilities. PMID- 14533052 TI - Service delivery options for secondary students with language disorders. AB - Numerous adolescents are still undetected, unserved, and underserved by speech language pathologists, resulting in astronomical financial and psychological costs to them and society. The purpose of this article is to provide an overview of past and present service delivery options for adolescents with language disorders. Before illustrating the authors' recommended model, the following background is provided: an overview of the three stages of adolescent development, characteristic expectations and problems for older students with language disorders, and a rationale for adolescent speech-language services. The comprehensive service delivery model for secondary-level speech-language students, designed and reported by the authors, consists of six components: information dissemination, identification, assessment, program planning, intervention, and follow-up. Some of the unique features of the comprehensive model are presenting services as a course for credit, providing grades, and using supportive course titles such as Individualized Communication Class. PMID- 14533053 TI - Reaching for the stars: SLPs shine on literacy teams. AB - Speech-language pathologists (SLPs) who work with school-age children have the opportunity to contribute to the literacy development of specific students as well as the literacy instruction and intervention programs in their district. These efforts are made in collaboration with other professionals and parents and may be referred to as participation on literacy teams. This article describes successful literacy teams in elementary schools including the variety of possible members as well as common characteristics of successful literacy teams. We describe how SLPs participate by developing collegial relationships with their team, using best practices for literacy, and supporting children as they progress in the general curriculum. Finally, we discuss some strategies for SLPs to take stock of their own literacy team participation, identify new goals, and map out a plan to reach new heights in literacy as integral members of literacy teams. PMID- 14533054 TI - An interdisciplinary team approach to providing dysphagia treatment in the schools. AB - The diagnosis and treatment of pediatric dysphagia in school systems are still in the infancy stages. They are being addressed in a variety of ways and by a variety of personnel. Speech-language pathologists (SLPs) in school systems throughout the country are faced with students who exhibit the signs and symptoms of dysphagia; however, there is a lack of organized procedures for the diagnosis and treatment of dysphagia in the schools. In some cases, SLPs are asked to treat a disorder for which they have no training. In both scenarios there are concerns about the services that students are receiving as well as professional liability. The purpose of this article is to address some of the more pressing issues related to the diagnosis and treatment of dysphagia in the schools and to provide the school-based SLP with some solutions to these problems. The question to be answered is, What is the best way to address dysphagia in the schools? PMID- 14533055 TI - Roles and responsibilities of the interpreter in interactions with speech language pathologists, parents, and students. AB - The Individuals with Disabilities Education Act Amendments of 1997 (IDEA '97) specify that students must be assessed in their primary language. With the ever increasing number of diverse languages spoken by students, it is very unlikely that even an English-bilingual speech-language pathologist's other language will match that of a student who is referred for a speech-language assessment. Knowing how to work effectively with an interpreter is, therefore, essential for all speech-language pathologists. This article reviews the process of interpretation during parent conferences and student assessments. It describes the specific roles and responsibilities of the speech-language pathologist-interpreter team, with a focus on the linguistic, cultural, and professional skills that the interpreter should have to ensure a successful outcome. Data obtained from bilingual Spanish/English-speaking speech-language pathologists are provided to confirm and describe some important features desired in an interpreter who collaborates with a speech-language pathologist. Suggestions for training and certifying future interpreters are outlined. Finally, models are presented for successful interactions with interpreters. PMID- 14533056 TI - Lessons from the National Outcomes Measurement System (NOMS). AB - Significant reforms in education are changing the way school-based speech language pathologists document and provide treatment. Faced with resource constraints and new mandates for accountability, clinicians are being pressured to demonstrate the value and impact of their services. To assist clinicians with meeting these challenges, the American Speech-Language-Hearing Association developed the National Outcomes Measurement System (NOMS) as a way to determine the effectiveness of school-based speech and language services. Data collected from NOMS are used to discuss: the effects of speech and language intervention on a student's ability to perform in the classroom, the key factors influencing progress for preschoolers with articulation disorders, and the impact of a clinician's caseload size on the outcomes students achieve. PMID- 14533057 TI - On "Minimal Pair Approaches to Phonological Remediation," (Semin speech lang 2002;23:57-68). PMID- 14533059 TI - [Cranial arteritis - An Overview]. AB - BACKGROUND: Cranial arteritis is one of the most important emergency situations in ophthalmology. METHOD: Recommendations of the literature how to diagnose and how to treat the disease are described. RESULTS: The basic diagnostic steps such as the "Five Criteria Classification" together with certain exceptions of the classification criteria and the recognition of different facets of the varying clinical picture are emphasised. Special arteritic appearances of the facial artery, the occipital artery and the vertebral artery should be noted and identified. It is pointed out that an arteritis without headache is of great clinical importance, known as occult arteritis, because of the lack of classical symptoms, often leading to a wrong diagnosis. Anterior ischaemic optic neuropathy (AION) is the most frequent cause of blindness. Differential diagnosis between non-arteritic and arteritic AION is emphasised. The different arguments concerning the performance of unilateral or bilateral arterial resection ("biopsy") and the findings of colour duplex ultrasonography in comparison to the histological results are discussed. It is also pointed out that some new diagnostic procedures such as the MRI examination with gadolinium and the PET Scan examination may be helpful in supporting the diagnosis. CONCLUSIONS: Cranial arteritis is more often diagnosed in recent years. The diagnosis should be made in cooperation by the ophthalmologist and internists and neurologists. Blindness caused by cranial arteritis is preventable in the majority of patients if diagnosed early and treated properly. For decades of clinical experience it has been well established that early diagnosis and systemic treatment with high doses of systemic corticosteroids are essential in preventing blindness in one or both eyes. PMID- 14533060 TI - [Clinical experiences and results of application of mitomycin C in trabeculectomy]. AB - BACKGROUND: Mitomycin C (MMC) used as an additive agent in glaucoma surgery has been shown to improve the postoperative results of intraocular pressure (IOP), but may also lead to higher incidences of postoperative wound healing disorders and of hypotonia with choroidal detachment. In this retrospective study the levels of IOP, the incidence of complications and changes of the visual acuity (VA) were monitored. METHODS: A trabeculectomy was performed on 70 eyes (57 patients). During surgery MMC was applied in three different concentrations: 0.1 mg/ml MMC (n = 8), 0.2 mg/ml MMC (n = 53) and 0.4 mg/ml MMC (n = 9) for 2.5 minutes, respectively. Patients were reexamined one day, and at 3 and 12 months after surgery. RESULTS: IOP was lowered with high statistical significance at all dates of control examination no matter what concentration of MMC had been used. At the time of discharge from hospital the success rate without additional topical glaucoma medication was 96 %, 78 % after 3 months and 68 % after 12 months. With glaucoma medication the success rate was 92 % after 3 months and 85 % after 12 months. Frequent complications were choroidal detachments in 40 % and hypotonia in 21 % of all cases. The VA did not change significantly during the follow-up period. CONCLUSIONS: This study underlines the efficacy of MMC as an additive agent in glaucoma surgery. However, considerable complications may be expected. Considering the effectiveness and the complication rate a concentration of 0.2 mg/ml MMC should be preferred. PMID- 14533061 TI - [Tetracaine drops versus lidocaine gel for topical anaesthesia in cataract surgery]. AB - BACKGROUND: in addition to intracameral anaesthesia, drop anaesthesia with tetracaine drops has been established for topical anaesthesia in cataract surgery. Anaesthesia with lidocaine gel is less established. We compared pain sensitivity of patients undergoing surgery with drop anaesthesia to patients who underwent surgery with gel anaesthesia. PATIENTS AND METHODS: In a questionnaire based survey in which patients assessed their sensitivity to pain during cataract surgery, we compared the sensitivity to pain of 49 patients during surgery with gel anaesthesia to 62 patients who consecutively underwent surgery with drop anaesthesia. Patients assessed pain intensity postoperatively by checking off a 10-graduation pain scale. Lidocaine gel (2 %) was applied once while tetracaine drops (1 %) were applied three times. All interventions were carried out by two surgeons as phako emulsifications with implantation of a posterior chamber lens via posterior-limbal access. In addition to pain sensitivity we also considered the patients' age, duration of surgery and surgical complications. The data for the patients' age, the duration of surgery and the assessment of intensity were not normally distributed so that a Mann-Whitney-rank-sum test was performed. RESULTS: No differences were observed between tho two groups, neither regarding age (P = 0.63) nor the duration of the surgical intervention (p = 0.43). However, in patients treated with tetracaine drops pain sensitivity was reported as being significantly less (mean 1,77) than in patients treated with lidocaine gel (mean 2.22; P < 0.043). 73.5 % of patients with gel anaesthesia and 83.9 % of those with drop anaesthesia reported "negligible" pains. There were no intra-operative complications. CONCLUSION: Both gel anaesthesia and drop anaesthesia are suitable methods for cataract surgery under local anaesthesia. According to patients' assessment, drop anaesthesia with tetracaine is better suited for pain elimination. PMID- 14533062 TI - [Shutter-glasses haploscope with eye-tracking control for fusion therapy]. AB - BACKGROUND: Common haploscopes require a manual input of the presented disparity and particularly prepared images; moreover, they can be used predominantly by orthoptists. We developed a device that makes possible a haploscope on the basis of a computer monitor with shutter-glasses. The presented disparity is determined with help of an eye-tracker. MATERIALS AND METHODS: A patient looks at a monitor through shutter-glasses, the eyes being recorded by a camera. An eye-tracking software determines the fixation points and the angle of strabismus. Images are presented at the monitor with a disparity that is regulated according to these measurements. This is controlled by means of a software so that the disparity is gradually decreased according to the patients abilities. Diagnostic data are recorded permanently. RESULTS: Any monitor contents, such as videos, TV or computer applications, can be used for fusion training or diagnostic purposes with the new apparatus. Initial investigations of patients with heterophoria or intermittent exophoria indicate a decrease of the subjective angle of strabismus by compensation and an increase of the fusional range. CONCLUSIONS: The developed shutter-glass-haploscope with eye-tracking control provides several methods for diagnosis. The appliance provides automated fusion training adapted to the patient that can be done at the working place or at home. PMID- 14533063 TI - [Flap Amputation in Mycobacterium chelonae Keratitis after Laser-in-situ Keratomileusis (LASIK)]. AB - BACKGROUND: Keratitis due to Mycobacterium chelonae after laser-in-situ keratomileusis (LASIK) is a rare, but severe complication. In the following report, we present clinical findings, microbiological investigation, treatment and outcome of the first case of Mycobacterium chelonae reported in Europe. PATIENT AND METHODS: A 52-year old woman presented with atypical unilateral keratitis after LASIK. Mycobacterium chelonae keratitis was diagnosed by microbiological investigation. Interface irrigation and treatment with topical and oral antibiotics was performed. RESULTS: Despite intensive treatment, flap removal was necessary to control the infection. Best-corrected visual acuity dropped from preoperatively 1.0 to postoperatively 0.2. CONCLUSION: The diagnosis of mycobacterial keratitis after laser-in-situ keratomileusis is often delayed due to atypical clinical appearance. Therefore consideration of atypical pathogens and rapid microbiological diagnosis is necessary to provide adequate treatment. PMID- 14533064 TI - [Cerebral atrophy in multiple sclerosis patients treated periodically with boluses of methylprednisolone]. AB - INTRODUCTION: In the last years advances in the treatment of the remittant and secondary progressive forms of the multiple sclerosis (MS) have taken place. In the primary progressive forms (PP) the side effects of potentially useful drugs prevent their use; in some studies an improvement of the evolution with the intravenous administration of periodic pulses of methylprednisolone (MP) has been observed. OBJECTIVE: To evaluate if periodic pulses of intravenous MP injected every 4-6 weeks increase the degree of cerebral atrophy of the patients with PP MS. PATIENTS AND METHODS: We studied 11 patients with PP MS treated during 33 months with periodic pulses of intravenous MP. The degree of cerebral atrophy was evaluated in axial cuts of 10 mm of studies of magnetic resonance with the indices of Evans, bicaudate, bifrontal, frontal spears and bithalamic. The statistical evaluation of the results was made applying the test of Wilcoxon-Mann Whitney. RESULTS: There are not statistically significant differences in the degree of cerebral atrophy with any of the used indices. CONCLUSIONS: Periodic pulses of intravenous MP do not accelerate the cerebral atrophy in patients with progressive forms of multiple sclerosis. PMID- 14533065 TI - [Posterior reversible encephalopathy in infancy]. AB - INTRODUCTION: Posterior reversible encephalopathy (PRE) is a radiological clinical syndrome that was initially reported in adults and which is characterised by seizures, headache, visual disorders, decreased awareness and anomalous neuroimages. It is an acute transient disorder. The most frequent causes are arterial hypertension (AHT) and immunosuppressive therapy. AIMS: The aim of this study is to describe the characteristics of PRE in patients of paediatric age. PATIENTS AND METHODS: From a total of 109,267 patients admitted to hospital between 1/1/1999 and 1/8/2002, we reviewed the case histories of seven patients who satisfied PRE criteria. RESULTS: Seven patients presented AHT associated with kidney disease (6) and with an undetermined causation (1). The initial symptoms were vomiting in seven and headaches in five patients, followed by seizures in six cases, which were partial (4), generalised (1), motor status (1). Five of them suffered visual disorders. All of them had decreased awareness and one of them required the assistance of mechanical ventilation. The condition cleared within 3 to 12 days after antihypertensive therapy. The anomalies that showed up in the neuroimaging studies (hypodensities in CAT, hyperintensities in T2 and hypointensities in T1 in MR) revealed the involvement of cortical and parieto occipital cortico subcortical regions asymmetrically. Frontal (2), temporal (2) and cerebellous (1) involvement was also observed. These disorders disappeared in patients from whom control images were obtained. CONCLUSIONS: PRE must be considered in the presence of the symptoms and the anomalous neuroimages described above, when associated with acute AHT. With correct management of arterial hypertension it is possible to clear the neurological symptoms and normalise the neuroimages that confirm this diagnosis. PMID- 14533066 TI - [Cerebrovascular disease in children with sickle cell anaemia]. AB - INTRODUCTION: Cerebrovascular disease (CVD) is one of the most clinically devastating complications in children with sickle cell anaemia (SCA) and an important cause of death in this population. AIMS: To determine the incidence of CVD in children with SCA and to characterise the risk factors and other related clinical and imaging aspects. PATIENTS AND METHODS: We conducted a retrospective, cross sectional, descriptive study of paediatric patients with SCA and CVD that covered the period between January 1992 and February 2002. RESULTS: A total of 131 children with SCA were registered in the period between January 1992 and February 2002, during which five children suffered some kind of CVD event. This figure corresponds to 3.81% of the patients with SCA and 7.69% of the total number of children with CVD. The most frequent clinical presentation of the CVD is bouts of seizures and the most frequently observed type of CVD is ischemic, chiefly located in the middle cerebral artery. Of the different predictors that exist for ischemic CVD, haemoglobin count was found to be lower than in haemorrhagic CVD. CONCLUSION: CVD is a common complication in the progress of patients with SCA with a high percentage of neurological sequelae. PMID- 14533067 TI - [Intracranial lipomas in paediatrics: a retrospective study of 20 patients]. AB - INTRODUCTION: Intracranial lipomas (ICL) are congenital malformations that are due to an anomalous differentiation of the primitive meninges. AIMS: The purpose of this study is to determine the most frequent symptoms related to ICL in the paediatric age, as well as to evaluate whether they are directly related to their location and to their association with other brain malformations. PATIENTS AND METHODS: A retrospective clinico-radiological study was conducted involving 20 paediatric patients diagnosed as suffering from ICL between 1985 and 2003 at three hospitals in the Valencian Community. RESULTS: 70% of the cases were females and the average age when ICL was diagnosed was 5.3 years (interval from 2 months to 13 years). The most common initial clinical justification for the neuroradiological study was psychomotor retardation in eight patients (40%) and persistent, predominantly migrainous, headaches in six of them (30%). Only one patient was examined because of epilepsy. The prevailing site was the interhemispheric fissure in 10 patients (50%), followed by the quadrigeminal cistern in five (25%) and the mamillary body infundibulum axis in three of them (15%). One case was located in the cerebellum and another was found in the lateral ventricle, with involvement of the choroid fissure. Associated anomalies were observed in eight patients (40%). Alterations were seen in the development of the corpus callosum in six patients (30%), five in an isolated way and in one case there was partial absence of the associated septum pellucidum. Another patient was found to have an isolated partial agenesis of the septum and Goldenhar syndrome was detected in another case. The radiological follow up, which varied according to the initial clinical features, did not show any differences in the morphology or the size of the lipoma. CONCLUSIONS: 1) ICL are more frequently located in the interhemispheric fissure and in the quadrigeminal cistern; 2) In these locations they can be associated with other brain malformations, the most important being defects in the callosa and septa; 3) Most ICL were found incidentally during the course of a neuroimaging examination. Only in the case of three patients (15%) was there any presumed relation between the location of the ICL (pericallosal), the symptomatology (overall psychomotor retardation) and the associated malformation (hypoplasia of the corpus callosum); 4) The incidence of epilepsy was much lower than that claimed in earlier reports, and the predominant associated neurological symptomatology was found to be psychomotor retardation and headache (70%); 5) ICL is a developmental anomaly that does not require neurosurgical treatment and which does not usually change with the passage of time, although it is liable to increase because of adipose hypertrophy. PMID- 14533068 TI - [Differences between migraine patients referred to a headache unit or a general neurology service]. AB - INTRODUCTION: Migraine accounts for 10% of patients first visits due to neurological reasons in Spain and over half the new visits in headache units (HU) and hence the importance of this pathology. AIMS: The aim of this study is to determine whether there are any differences between migraine patients referred to a general neurology service (GNS) or to a HU. PATIENTS AND METHODS: Two groups of patients with migraine were compared: those sent for the first time to a GNS and the others, who were sent directly to a HU. RESULTS: In a GNS, 10.7% (374 patients) of the overall number of new visits concerned migraines; these were compared with 107 migraines (64%) from the total number of headaches treated for the first time in the HU during the year 2000. The average age and distribution of sexes were similar in both groups. In the group of migraines from the HU there were more requests for CAT/MRI (20%), 77.5% had previous treatment, 71% were given preventive therapy, 51% received triptans and 44.8% needed an examination. In the group of migraines from visits to general service, there were fewer requests for CAT/MRI (14%), only 20% had previous therapy, preventive therapy was started in 45%, 6% received triptans and 25% required an examination. CONCLUSIONS: The group of patients with migraine who were sent to the HU presented a more serious pathology, required more preventive therapies, more triptans and more monitoring than the group of patients with migraine referred to the GNS. PMID- 14533069 TI - [Mucopolysaccharidosis type I in the Cuban population]. AB - INTRODUCTION: Of all the innate errors of the metabolism, the mucopolysaccharidoses (MPS), a kind of lysosomal disease, are especially significant because of the serious clinical features they give rise to and the therapeutic difficulties they entail. Diagnosis of the index case is essential so that families can gain access to the preventive benefits of genetic counselling. To date, seven types of MPS and 11 enzyme deficiencies have been described. AIMS: Patients with a clinical diagnosis that leads the clinician to suspect they may be suffering from some type of MPS are referred to the Institute of Neurology and Neurosurgery to determine the possible enzyme deficiencies that will provide us with the key to a successful diagnosis. PATIENTS AND METHODS: A total of 588 patients who were clinically suspected of suffering from MPS, in whom 1,502 enzyme analyses were performed in order to classify the type of MPS they were suffering from. The MPS under examination are MPS I, MPS IIIB, MPS IVB, MPS VI and MPS VII. RESULTS: MPS I, or alpha-L-iduronidase deficiency, is the most commonly found with 23 cases (4.08% of the patients studied); 13 were females and the other 10 were males. Of the 23 cases, 10 presented the severe Hurler phenotype with mental retardation, five had the Scheie phenotype with preserved intelligence and eight displayed the intermediate Hurler-Scheie phenotype. Diagnosis was reached before the end of the first year in eight patients, between 1 and 5 years in nine of them and between 6 and 10 years in two cases. Enzyme activity in leucocytes was significantly lower in patients as compared to a control group and with respect to the parents (heterozygotes), and even comparing these to the control group, with a slight and expected overlap. CONCLUSION: The biochemical methodology used allows us, then, to reach a sure biochemical diagnosis and to offer the families the benefits of genetic counselling. PMID- 14533070 TI - [Treatment of a case of generalised dystonia using subthalamic stimulation]. AB - INTRODUCTION: Generalised dystonia is an entity that does not usually respond well to medical treatment. Different surgical targets have therefore been used in the treatment of dystonia, including several thalamic nuclei or the internal globus pallidus. The subthalamic nucleus plays a fundamental role in the physiology of the basal ganglia. It could therefore be considered to be a good potential target for stimulation. CASE REPORT: A patient who was confined to a wheelchair and who had not responded to a number of different medical treatment protocols or to a bilateral thalamotomy was treated with bilateral deep brain stimulation in the subthalamic nucleus. Tetrapolar electrodes were placed in both subthalamic nuclei in two stages. The patient showed a significant improvement from the very beginning of the post-operative period. After six months' progression, the patient was able to walk unaided and the dystonic seizures diminished significantly. Unfortunately, the patient died from choking. We used a bipolar stimulation protocol at 50 Hz with 210 micros pulses, which do not reach the levels of maximum charge density that are considered to be harmful. CONCLUSION: The subthalamic nucleus can be a good surgical target for deep brain stimulation in cases of generalised dystonia; it responds well to stimulation at intermediate frequencies with safe charge densities. PMID- 14533071 TI - [Multiple cerebral tuberculomas]. AB - INTRODUCTION: The tuberculosis is a disease that continues being important cause of morbidity and mortality at worldwide level. Its presentation as tuberculomas cerebral manifold at level of the central nervous system is little frequent in immunocompetent patients and can be confused with other etiology. CASE REPORT: An indigenous young man, immunocompetent consulted for history of headache, nausea, vomits, convulsions, double vision and hemiparesia left side, which in the cerebral tomography of revenue was showing injuries compatible with cerebral abscesses; for which he received treatment with antibiotics without improvement for what there takes biopsy of the injuries that reported tuberculomas, specific treatment being initiated later and the primary area being investigated without the same one be detecting. After the first procedural step with evident clinical and radiographic improvement. CONCLUSIONS: The tuberculosis in anyone of their forms of presentation must be included within the diagnosis differential of the patients in our endemic countries for this disease. The clinical and radiological diagnosis of cerebral injuries is difficult and single usually it obtains to the diagnosis during a pathology study that shows tuberculomas with caseosa necrosis, epiteliodes cell and the acid alcohol bacilli resistant. PMID- 14533072 TI - [Progressive myoclonic cerebellar ataxia as a manifestation of Creutzfeldt-Jakob disease]. AB - INTRODUCTION: Progressive myoclonic cerebellar ataxia is a clinical entity with an important spectrum of possible diagnoses that requires a complex and exhaustive differential diagnosis. CASE REPORT: A 53-year-old male patient with no relevant medical history who was admitted to hospital because of an unstable gait, together with mild bilateral dysymmetry and the progressive and insidious widening of the base of support. The patient's symptoms then became more pronounced and included myoclonus and a deterioration of the higher functions. The patient died four months after the onset of the symptoms. The explorations that were conducted included a pathological study of the brain, which confirmed the diagnosis of classical spongiform encephalopathy (Creutzfeldt-Jakob disease). CONCLUSIONS: Creutzfeldt-Jakob disease must be included in the differential diagnosis of progressive cerebellar ataxias. PMID- 14533073 TI - [Calculating how long phenytoin intoxication lasts]. AB - INTRODUCTION: We report a case of intoxication with phenytoin (DPH), in which the actual time required for it to disappear was compared with that estimated using linear regression. CASE REPORT: A 23-year-old female with tonic-clonic seizures, receiving chronic therapy with DPH 300 mg/day. The patient came to hospital because of tremors, balance disorders, vomiting and headaches. Neurologically, she presented horizontal nystagmus in the two extreme gazes, generalised hyperreflexia and ataxic gait. Cranial CAT scan and cerebrospinal fluid were both normal. Serum concentration of DPH was found to be 60.2 mg/L. When DPH concentration is >8-10 mg/L, its rate of elimination diminishes disproportionately and the risk of toxicity increases. Use of mathematical methods makes it possible to calculate the time required for a toxic concentration to come down to therapeutic values. In our patient the DPH took 204 hours to drop below the toxic level (20 mg/L), whereas by using a linear regression with only two different concentrations a figure of 155 hours was obtained. CONCLUSIONS: The method employed here can be useful as a quick, simple and easily applicable way of estimating the time a toxic concentration of DPH takes to return to a normal level. PMID- 14533074 TI - [An association between moya-moya disease and morning glory anomaly]. AB - INTRODUCTION: The morning glory disc anomaly is a not very frequent congenital alteration of the optic papilla associated with a retinal vascular dysgenesia. Several cases of association have been described between this disc anomaly and the presence of Moyamoya vessels. We report the case of a boy with this association. CASE REPORT: A 5 year-old boy was evaluated for several episodes of disconnection, some of them preceded by intense headaches and associated to incoherent language, and in other occasions with hypotonia and unmotivated laugh of variable duration. The physical and neurological exploration was normal, except for the optic disc papilla of the right eye that was very big and dug, compatible with Morning glory disc anomaly. No alterations were found at cranial CT and EEG. In the magnetic resonance imaging were visualized multiple small round low intensity areas in the basal ganglia, because of the high number of collateral vessels. The angioRM showed stenosis of the supraclinoid portions of both internal carotids as well as the proximal segments of the anterior cerebral artery and the left middle cerebral artery with abnormal network of collateral vessels at the base of the brain like a puff of smoke . This is a typical imaging for Moyamoya disease. CONCLUSIONS: In all patient with Morning glory disc anomaly, mainly if they presents neurological clinical features, the realization of angioRM studies will be evaluated, to discard the existence of Moyamoya disease. PMID- 14533075 TI - [Experimental approaches to perceptual dysfunction in schizophrenia]. AB - METHOD: Starting from the studies of Conrad on early schizophrenia, we quantified the gestaltic alteration that he has described using a visual-motor gestaltic test (Bender test) in acute and chronic schizophrenic patients. We observed that rotations and distortions were significantly higher in chronic patients, and perseverations in acute patients. Global scores and time employed showed significant differences in both groups when compared with controls. Time is classically considered a compensation in the failure. We observed that the Bender gestaltic test allows quantification of perceptual alterations due to the loss of the objective structure of the perception in schizophrenic patients. Aiming to reproduce these findings in an animal model, we proposed a study of pharmacological modification of nucleus accumbens septi (Acc) neurotransmission, traditionally linked with physiopathology of schizophrenia. In this way, we developed a discrimination shape model in a skinner box in trained pigeons, bilaterally implanted in Acc by stereotaxic surgery. CONCLUSIONS: The antagonists of N-methyl-D-aspartic acid (NMDA) glutamatergic receptor induced a significant decrease in performance in the discrimination task, and a significant increase in correcting trials. The last parameter was considered a perseveration, manifestation of a behavioral inflexibility in relationship with environmental changes. The glutamatergic blockade of Acc in rats using a passive avoidance task induced a disturbance in acquisition, and the same procedure in the plus maze test led to a significant decrease in anxiety levels, suggesting additional homologies with schizophrenic psychoses. PMID- 14533076 TI - [Palliative surgical treatment of spastic paralysis in the lower extremity]. AB - AIMS: The purpose of this paper is to describe our experience with and to review the best results in the surgical treatment of patients suffering from spastic paralysis of the lower limbs. DEVELOPMENT: To enable a correct indication of the techniques to be employed the authors recommend a thorough examination of the types of deformity (fixed, dynamic or mixed) and the use of specific tests for exploring the different deformities. These are necessary steps to be able to interpret the different disorders in a global manner and thus reach diagnostics that provide us with a proper surgical therapeutic orientation about the spastic hip, knee, ankle and foot. Due to the importance of the overall problem, it is becoming increasingly more frequent to advise multidisciplinary work involving the collaboration of different specialists (neurologists, rehabilitators, physiotherapists, psychologists, paediatricians, neurophysiologists and orthopaedic surgeons). Spasticity is as heterogeneous as the results of the different treatment projects. The techniques used must allow the rehabilitation therapy to be continued. Surgical intervention is recommended when the damage to the CNS has stabilised and the patient is over 4 years old. The psychic state of the patient and the family must also be evaluated. CONCLUSIONS: The objective of the treatment in patients who can walk is to improve motor functioning, the type of gait and to prevent fixed deformities from developing. In patients who do not walk, the aim is to improve their hygiene and their capacity to sit and to walk. These indications are indispensable to be able to successfully perform a little known area of orthopaedic surgery which does not respond to the techniques used in flaccid paralysis surgery. PMID- 14533077 TI - [Levetiracetam in children and adolescents with epilepsy]. AB - AIMS: The aim of this paper is to report on the clinical experiences that have been published concerning the association of levetiracetam (LEV) in children and adolescents with refractory epilepsies. DEVELOPMENT: Although LEV has been approved for use in polytherapy in patients over the age of 16, some short works (communications and posters in international congresses) and three open studies have already dealt largely with the fact that the drug has been associated in children and in adolescents under the age of 16, with all types of refractory seizures, especially focal seizures. Half the patients can be considered to be responders, since the seizure rate is reduced by more than 50% when LEV is associated, and were completely eradicated in 16% of cases. In a third of the cases side effects were reported, although they can be seen as being mild and transient since they meant that LEV application was interrupted in less than 10% of cases. CONCLUSIONS: LEV is a drug that is effective and well tolerated in children and adolescents with difficult-to-treat epilepsies; it is very straightforward to manage due to its excellent pharmacokinetic characteristics and suggested dosages at these ages are between 40 and 50 mg/kg/day (up to 3,000 mg per day, in two doses). PMID- 14533078 TI - [The development of attention, memory and the inhibitory processes: the chronological relation with the maturation of brain structure and functioning]. AB - Development during childhood and adolescence is characterized by greater efficiency in performing cognitive tasks. The correlations between cognitive and brain development are not altogether clear and have not been studied in depth. The aim of this study is to survey the research carried out into the development of cognitive functioning in children, adolescents and adults, and its chronological relation with brain development. Anatomofunctional and cognitive behavioural studies are presented. Anatomical studies have shown that the white matter increases linearly throughout childhood and adolescence, whereas cortical and subcortical grey matter increases in the pre-adolescent period and later diminishes in the post adolescent stage. It has been claimed that these changes are regional and that the prefrontal cortex (PFC) is one of the last areas to mature. Functional research has studied cognitive processes attributed to the functioning of the PFC, such as attention, working memory and the inhibition of irrelevant responses. The findings from these studies have shown a behavioural and physiological development of these three processes during childhood and adolescence. Behavioural results have evidenced greater efficiency in capacities such as discriminating between relevant and irrelevant information, storing and handling information in the memory and the inhibition of unsuitable responses during the performance of a task. The physiological results have presented changes in the magnitude, spread and integration of the regions activated during task performance. Cognitive and behavioural maturation is consecutive to structural and physiological maturing and this is produced in a chronologically and qualitatively different way in the distinct regions of the brain. PMID- 14533079 TI - [Applications of functional magnetic resonance imaging in pre-surgical patients: motor, mnemonic and linguistic functions]. AB - AIMS: This paper reviews and describes cases involving useful protocols for pre surgical evaluation in functional magnetic resonance imaging (fMRI) of the motor, mnemonic and linguistic functions. DEVELOPMENT: The application of fMRI in pre surgical evaluation allows us to pinpoint the anatomical location of a function in relation to the lesion. This technique therefore appears as an alternative to the classical intraoperative tests, which are more aggressive and costly. The paper describes aspects and considerations of fMRI with regard to the methodology and procedure employed in its application, as well as the control of contaminating variables that can affect results. The language protocols are described in terms of lateralisation and localisation, and are divided into expressive and receptive language protocols, together with linguistic protocols for overall stimulation that allow the localisation of both expressive and receptive areas. Concerning the motor protocols, we describe the procedures for patients with and without difficulties in their movement. Lastly, we review and describe the memory protocols that give rise to bilateral stimulation in the mesial temporal areas, which allows both lateralisation and localisation of the hippocampal and parahippocampal areas that are functionally significant for the patients' memory. Each of the protocols is illustrated with cases carried out by our work team. CONCLUSIONS: The development of new protocols for pre-surgical evaluation must be guided by greater delimitation and consistency in functional stimulation, as well as analysis of the behaviour displayed. PMID- 14533080 TI - [Mid-latency auditory evoked potential]. AB - AIMS: Given the diversity of criteria regarding the use of the midlatency auditory evoked potential (MLAEP) for diagnostic purposes, we carry out a review of the subject and highlight areas such as the generators of fundamental components, the conditions in which responses are obtained, factors that can modify results and the clinical uses that have been reported. DEVELOPMENT: There seems to be a strong tendency towards considering areas of the primary and secondary auditory cortex as generators of the response, with the subcortical structures playing a moderating role. Factors such as age, the method of stimulation used and the monitoring conditions can all exert an influence on the characteristics of this evoked response and modify its capacity for detection and repetition. Clinical uses of this potential range from the detection of the hearing threshold, monitoring the depth of anaesthesia, the functional evaluation of cochlear implants, and the diagnosis of neurological disorders, among others. CONCLUSIONS: MLAEP is a technique that is widely used in the study of the most rostral segments of the auditory pathway and this information complements that provided by short latency responses and increases the diagnostic sensitivity of auditory evoked potentials. PMID- 14533081 TI - [Hypertension, cardiovascular reactivity to stress and sensibility to pain]. AB - OBJECTIVE: To provide a review of empirical evidence of decreased pain perception in hypertensive persons or exaggerated cardiovascular reactivity to stress. DEVELOPMENT: To following article will briefly review the existing literature on the association between hypoalgesia and high blood pressure. In particular, evidence of hypoalgesia in normotensive individuals at increased risk for hypertension (exaggerated cardiovascular reactivity to stress) will be offered in support of the notion that high cardiovascular reactivity to stress and decreased pain perception may result from a common physiological dysfunction. Cardiovascular reactivity refers to changes in cardiovascular activity associated primarily with exposure to psychological stress. Different individuals show different amounts of reactivity under the same conditions. The greater cardiovascular reactivity to behavioral stressors may play some role in the development of sustained arterial hypertension. Central opioid hyposensitivity is hypothesized as a mechanism of both hypoalgesia and exaggerated autonomic and neuroendocrine responses to stress in individuals at risk for hypertension. The paraventricular nucleus of the hypothalamus (PVN) serves the crucial function of integrating cardiovascular and painful responses. The central opioid hyposensitivity model of hypoalgesia asserts that attenuation of inhibitory opioid input to the PVN may have important consequences for pain modulation. These consequences includes: 1) greater activation of baroreceptor reflex arcs, 2) enhanced release of endogenous opioids during stress, and 3) increased stimulation of descending pain modulation pathways. CONCLUSIONS: High elevated thresholds to painful thermal stressors might serve as a behavioral marker of risk for hypertension before the onset of high blood pressure levels. PMID- 14533084 TI - [Somatosensory evoked potentials in decompression sickness]. PMID- 14533085 TI - [Topiramate therapy in patients with refractory epilepsy]. AB - AIMS: The purpose of this study was to examine the result of administrating topiramate (TPM) to patients with epilepsy that is refractory to treatment with two or more antiepileptic agents. PATIENTS AND METHODS: A total of 50 patients were evaluated, 90% of which had partial seizures (simple partial, complex partial and partial with secondary generalisation) and the remaining 10% suffered from generalised seizures. The most frequent aetiology was that which corresponded to the symptoms (52% of the cases). 98% of the patients were treated with two or three drugs. TPM was added with a dosage interval between 75 and 550 mg and follow-up visits were carried out throughout a period of nine months. RESULTS: 12 patients remained seizure-free for at least the first three months of the study; in 20 patients the number of seizures decreased by 50% or more; in 14 patients no changes were observed, and in the other four there was an increase in the number of seizures. The best results were obtained in cases of seizures with an idiopathic aetiology, while complex partial seizures offered the worst results. We also analysed the factors that might exert an influence on the different degrees of response. CONCLUSIONS: TPM was effective in all kinds of seizures. 24% of the patients became seizure free, and the total percentage of respondents rose to 64%. No patients dropped out of the study because of intolerance to the drugs. PMID- 14533086 TI - [Subthalamic nucleus targeting and spatial variability]. AB - AIM: The effectiveness of anatomic localization of the subthalamic nucleus (EAL) was assessed and the mapping method is described here. The symmetry of contralateral nuclei (SCN) was analyzed on 11 parkinsonian patients submitted to bilateral subthalamotomy with ablative lesioning. PATIENTS AND METHODS: To assess EAL the percentage so much of first trajectory (p1) as the total of trajectories (pt) that hit the target and the rest of subthalamic nucleus average distance (d) was calculated. The anatomic localization error (epsilon) is determined as a difference between first trajectory coordinates with those of medial determined nucleus point, through electrophysiological data as to the statistical significance of this error. SCN is analyzed by contrasting equality hypothesis at the nucleus maximum height alongside a trajectory, average electrophysiological position center and spatial distribution of all intranuclear recordings found in each hemisphere in all patients. RESULTS: The pi, pt and d obtained values were 86.36%, 86.13% and 1.41 +/- 1.01 mm respectively. The epsilon value was greater in anteroposterior direction of 1.11 +/- 0.83 mm without statistical significance. The average number of recorded trajectories for the first procedure was 6.45 and 6 for the second. The asymmetry of contralateral nucleus was not significant. CONCLUSIONS: An indirect method with CT brain images and a new electrophysiological mapping method with a multiunitary recording for first and second nucleus is safe enough and it yields a high effectiveness in anatomofunctional nucleus localization. The nucleus of a same patient are symmetrical. There is little space variability among patient non related to the differences in the intercommissural distance. PMID- 14533087 TI - [Neonatal convulsions: influence of the electroencephalographic pattern and the response to treatment on the outcome]. AB - INTRODUCTION: Neonatal convulsions continues being motive for multiple controversies: the diagnosis only by clinical approaches, the necessity to confirm with EEG record and their treatment and control. OBJECTIVE: To establish the incidence of type of clinical neonatal seizures and the correspondence of these to the EEG trace background and the EEG epileptic activity, the underlying etiology, the response to antiepileptic treatment, and their prognosis. PATIENTS AND METHODS: Retrospective study of admitted newborns with the diagnosis of neonatal seizures in our hospital, during the period included between January 1993 and October 2001. Some of variables collected were: birth and gestational age, Apgar, clinical pattern, duration of the convulsions, critical and intercritical EEG traced, EEG background, etiological diagnosis, treatment used and response of the same, evolution and neurological state at hospital discharge and at one year of age (corrected age in preterm infants). RESULTS: 74 children were admitted with neonatal convulsion diagnosis, of these only 56 EEG convulsions were confirmed (42% presented subtle seizures, 33.9% tonic, 64.3% clonic multifocal, 10.7% clonic focal, and 16.1% myoclonic multifocal). 55.4% of the infants had 2 or more types of clinical convulsions, 25% of all had an epileptic state, and 42.9% had at some time of the EEG record, electroclinical dissociation. The more frequent critical EEGs abnormalities was multifocal discharges (64.3%), and together with the focal discharges of low frequency had significant (p<0.01) worse pharmacological control, and also unfavourable outcome. The infants having had EEGs background moderately and markedly abnormal showed unfavourable outcome in 72.2% and 100% respectively, while it was only in 15.4% of the infants who had EEGs background normal or lightly abnormal. With the antiepileptic treatment the clinical control of the convulsions was obtained in more than 80% of the cases, while control of the electrical convulsions was only in 62.5%. There was a higher significant association between favourable response to treatment and normal neurological examination at hospital discharge and at 1 year of age. CONCLUSIONS: The necessity to confirm by means of EEG record the neonatal clinical convulsions, before and after having established the anticonvulsant treatment, due to the control of electrical convulsions improves their neurological outcome. PMID- 14533088 TI - [The control of somesthesia caused by neuroreflex stimulation: quantification of stimulus response as opposed to energy]. AB - INTRODUCTION: In the reflexotherapy of classical acupuncture it is usually claimed that below a certain level of stimulation, which is determined by the appearance of a peculiar sensation known as De-Qi, no appreciable therapeutic effect is obtained. The control exerted by the afferences over the central nervous system, however, do not work in this way, but instead adjust themselves, albeit with certain peculiarities, to a classical pattern of proportionality between stimulus and response. Our aim is, therefore, to evaluate the extent to which increasingly intense acupuncture stimuli can modify neurophysiological parameters in a way that also grows. We will focus on the quantification on a metametic level, through the study of the F-wave, and in the telencephalon, through somatosensory evoked potentials. PATIENTS AND METHODS: A total number of 21 healthy volunteers were studied and we collected the values corresponding to the parameters indicated in the basal situation, when needling an extra acupunctural point and following needling of the classical point IG4 until the sensation of De-Qi was felt, and electrostimulation was added after detecting the abovementioned sensation. RESULTS: We observed a clear progression in the modifications of both the F values and the somesthetic potential, with growing stimuli. Basic inflection does indeed occur after obtaining De-Qi, but on overstimulating from then onwards greater variations are produced. This is especially obvious in the case of the latency of the somesthetic potential. Needling a point that is not on a meridian guarantees that merely piercing the skin does not exert any of the effects mentioned above. DISCUSSION: In spite of the classical postulates, it can be observed that the more intense the acupuncture stimulus is, the greater the modifications to the neurophysiological parameters being studied are. The control of the somesthetic afference by acupuncture is something that has already been demonstrated and quantified. What is now seen, in addition, is the direct relation between the intensity of that effect and that of the stimulus used to produce it, which is quite different from the classical conceptions of energy. CONCLUSIONS: The effect of acupuncture stimulation is closely related to the intensity of the stimulus applied; the sensation of De-Qi is not necessary to obtain neurofunctional modifications and stimulations that go beyond it cause greater effects. PMID- 14533089 TI - [First episode non-traumatic subarachnoid haemorrhage. An epidemiological study of incidence and of arterial hypertension as a risk factor]. AB - AIMS: Our aim was to conduct an epidemiological study of the incidence of non traumatic subarachnoid haemorrhage (NTSAH) and to evaluate a history of arterial hypertension (AHT) as a risk factor (RF). We also sought to determine the gross annual rate (GAR) and specific rates, by sex and age, of first episode non traumatic subarachnoid haemorrhage (FENTSAH) adjusted to the Censo Nacional de Poblacion de la Argentina (CNPA), and to carry out a population-based case control study about the history of AHT as a RF in an Argentine community. PATIENTS AND METHODS: Our institution, the Hospital Privado de Comunidad de Mar del Plata, Argentina, attends a population of about 89,500 individuals exclusively and completely. There is a single filing system of case histories with all the diagnoses in a database. Patients who suffered a FENTSAH between the years 1992 and 1996 (n=62) were investigated to determine the specific GAR by sex and age and adjusted to the 1991 CNPA. The population was compared at random with two controls per case for the study of a history of AHT as a RF, and was then stratified according to sex and age. RESULTS: 1. GAR: 13.6/100,000 inhab./year; 2. Specific rate for males: 10.5/100,000 inhab./year; 3. Specific rate for females: 15.9/100,000 inhab./year; 4. CNPA adjusted rate: 7.3/100,000 inhab./year; 5. A significantly important number of patients with FENTSAH presented a history of hypertension as compared to controls subjects (odds ratio: 4.73; CI 95%: 2.39 9.34). CONCLUSIONS: Our rates are not substantially different from the rates found in other western countries and our findings indicate that a history of AHT is a RF for FENTSAH. PMID- 14533090 TI - [A report of 62 cerebral aneurysms that were clipped at the Hospital Regional Docente Vicente Corral Moscoso in the city of Cuenca, Ecuador]. AB - AIMS: The purpose of this study was to establish certain epidemiological characteristics of cerebral aneurysm surgery at the Hospital Regional Docente Vicente Corral Moscoso (HVCM), in the city of Cuenca. PATIENTS AND METHODS: We examined the medical records of the patients who were treated at the HVCM, in Cuenca, over a 14 year period after being diagnosed as suffering from aneurysmal subarachnoid haemorrhage (ASAH), which was confirmed using cerebral arteriography. Patients were catalogued according to age, sex, origin, marital status, the number of months the aneurysm had been ruptured, their Hunt and Hess score on admission, incurrent diseases, habit of smoking, neurological signs on admission, the site of the aneurysm, treatment received, complications, surgery rupture and admission-surgery time and outcomes when discharged. Only patients who had undergone surgical treatment were considered. RESULTS: There were an average of 6.07 cases of ASAH and 4.7 aneurysm operations a year. Most patients were females from the provinces of Azuay and El Oro; the average age was 45.9 years, and incidence was higher in the months of March-April and August September. The most common site for presentation was the posterior communicating artery, which occurred in 44% of the cases. The most frequently observed preoperative complications were rebleeding and vasospasm. Surgery was performed on a total of 67 patients and none of them were submitted to early surgery. Operative death rate was 6%. Overall results were optimal in 55.2%. CONCLUSIONS: The therapeutic option offered by our service for this pathology is satisfactory. Indeed, it is similar to that employed in other centres throughout Latin America and has improved over the last few years thanks to the experience that has been acquired in this field. PMID- 14533091 TI - [Duplication of the PLP gene and the classical form of Pelizaeus-Merzbacher disease]. AB - INTRODUCTION: Pelizaeus-Merzbacher disease (PMD) is a rare form of sudanophilic leukodystrophy which is transmitted by recessive inheritance linked to the X chromosome. It only affects the myelin of the central nervous system (CNS) and is caused by a proteolipid protein (PLP) deficit, which is coded for in Xq21.2-q22. Presentation follows a classical or connatal pattern and is associated with nystagmus, stridor and pyramidal/extrapyramidal manifestations within the framework of a clinical picture of psychomotor retardation and regression with variable clinical course and presentation. CASE REPORT: A 37-month-old male, with sever psychomotor retardation, nystagmus and choreoathetotic movements with a stationary developmental profile. An MRI scan of the brain showed severe supratentorial hypomyelination and peripheral electrophysiological explorations (EMG and NCS) were normal. The genetic study using PCR revealed duplication in the PLP gene. CONCLUSION: This observation corresponds to a classical form of PMD, which must be taken into account when associated with: 1) Psychomotor retardation; 2) Early nystagmus; 3) Pyramidal/extrapyramidal involvement; 4) Absence of peripheral neurophysiological involvement; 5) A neuroradiological pattern of hypomyelination of the CNS. PMID- 14533092 TI - [The onset of an anaplastic ganglioglioma during the post-natal period with signs of toxemia of pregnancy]. AB - INTRODUCTION: Gangliogliomas are infrequent neuronoglial tumours which present in youngsters and are usually located in the temporal lobe. They usually appear with epileptic seizures and prognosis after surgical excision is usually good. The anaplastic forms are even less frequent and prognosis is poorer. The onset of epileptic seizures during the early post-natal period means that the clinician has to resort to a broad differential diagnosis. CASE REPORT: Hours after a preterm birth, at the 32nd week of gestation, a 35-year-old primipara began to suffer seizures and also presented arterial hypertension, proteinuria and a low platelet count. A cranial computerized tomography scan was carried out where a left frontal hypodense lesion was observed. Transcranial echo Doppler scan showed medium speeds and suggested eclampsia. The seizures, however, recurred during the days that followed and a magnetic resonance scan of the head revealed a lesion with nodular contrast enhancement, which was excised, and finally an anatomopathological diagnosis of an anaplastic ganglioglioma was reached. DISCUSSION: The toxemia of pregnancy, which gave rise to a vasogenic cerebral edema, accelerated the clinical onset of a brain tumour during the post-natal period. A ganglioglioma, although infrequent, is always a possibility to be borne in mind in young patients. PMID- 14533093 TI - [Hyperhomocysteinemia as a risk factor for central retinal vein thrombosis in a young patient]. AB - INTRODUCTION: Central retinal vein thrombosis (CRVT) is a multifactorial disease and known risk factors include high blood pressure, diabetes mellitus, hypercholesterolemia and primary open-angle glaucoma. Yet in patients below the age of 60, a state of hyperviscosity and hypercoagulability in the absence of other vascular risk factors are important factors, as is hyperhomocysteinemia, an independent risk factor for venous thromboses. Homocysteine is an essential amino acid produced by the transmethylation of methionine. It can be remethylated using enzymes that require folate and cobalamin to re-form methionine or to catabolize for cystathionine beta-synthetase, which is dependent on pyridoxine to form cysteine. Hyperhomocysteinemia can give rise to a dysfunction in the endothelium of the vessel, with a proliferation of vascular smooth muscle and prothrombotic homostatic changes. CASE REPORT: A male, aged 33, with no vascular risk factors except cigarette smoking who presented a sharp drop in visual acuity in the right eye. An ophthalmological examination revealed the presence of CRVT, which was confirmed by fluorescein angiography. The results of all analytical and imaging studies conducted while the patient was in hospital were negative. Later analytical monitoring showed a moderate increase in homocysteine. CONCLUSION: From a survey of the literature we have found an important relation between CRVT and hyperhomocysteinemia, which has been noted as a potential risk factor and which requires therapy. PMID- 14533094 TI - [Iatrogenic botulism: a complication to be taken into account in the treatment of child spasticity]. AB - INTRODUCTION: During the last decades the use of botulinum toxin for management of muscular disorders and spasticity associated to cerebral palsy has become a widespread practice. CASE REPORT: A 6-years female suffering of cerebral palsy secondary to a partial agenesis of the corpus callosum who was receiving bolulinum toxin since October 2001. One week after the last dose (Dysport 46 U/kg/dose) she started high grade fever, malaise, food refusal, choking, constipation, eyelid ptosis, absence of deep tendon reflexes, and abundant mucous discharge. Such features were so progressively severe that ventilatory support became mandatory. After a previous dose five months before, she developed similar features but they were less severe and thought to be related to a respiratory infection. CONCLUSIONS: At the present, there are two forms of botulinum toxin commercially available: the British brand Dysport and the American brand Botox. Bio equivalences are 1 U Botox to 2 or 6 U Dysport. Dosing (U/kg per session) has been established as follows: 5 U for Botox, 35 U for Dysport. Safety limits are a wide range with a therapeutic toxic index rate of 1:10. Generalized side effects after diffusion to blood of locally injected botulinum toxin are rare. PMID- 14533095 TI - [Cerebellar ataxia and absence epilepsy: genes, channels, neurons and mice]. AB - AIMS: To briefly review the most significant progress made in the fields of molecular genetics and cellular electrophysiology that have contributed to reveal that some paroxysmal disorders including epilepsy, are linked to voltage gated Ca2+ (CaV) channel abnormalities. DEVELOPMENT: Recent studies showed that the gene encoding the P/Q type Ca2+ channel is altered in a number of mutant mice strains. Phenotypic alterations of these mice include cerebellar dysfunction and absence epilepsy. CONCLUSIONS: These mutant mice offer a unique opportunity to elucidate the molecular, developmental, and physiological mechanisms underlying CaV channel activity in the brain, and provide the possibility to link specific CaV channel subunits to cellular disease processes, including altered excitability, synaptic signaling, and cell death. PMID- 14533096 TI - [Palliative surgical treatment of spastic paralysis in the upper extremity]. AB - AIMS: In this paper we review the main studies conducted on therapy applied to the bony and soft parts in spastic paralysis of the upper extremity. DEVELOPMENT: Spasticity presents muscular hypertonia and hyperexcitability of the stretch reflex, which are typical of upper motoneuron syndrome. Physiopathologically, spasticity is due to the medullar and supramedullar alteration of the afferent and efferent pathways. Treatment is multidisciplinary and involves the collaboration of rehabilitators, neurophysiologists, neurologists, paediatricians, orthopaedic surgeons and psychologists, who all contribute with their different therapeutic aspects and characteristics (which can be pharmacological, peripheral neurological blockages, surgical, etc.). The characteristic posture of the upper extremities in spastic cerebral palsy is the inward rotation of the shoulder, flexion of the elbow and pronated forearm, and the deformity of the fingers (swan-neck and thumbs-in-palm). The primary objectives in these patients will be to improve communication with their surroundings, perform activities of daily living, increase mobility and walking. CONCLUSIONS: The surgical treatment applied by orthopaedic surgeons in the upper extremities are aimed at achieving an enhanced adaptive functionality rather than morphological normality. Factors to be taken into account include age, voluntary control over muscles and joints, level of severity of the spasticity (Ashworth scale) and stereognostic sensitivity. In general, on soft parts we will use procedures such as dehiscence or lengthening of the flexor muscles of the shoulder and elbow or of the adductor of the thumb; transfer of the pronators in order to adopt the supinating function or of the flexors so as to reinforce the extensors of the forearm, and capsulodesis or tenodesis in the hand. The bony procedures will consist in derotational osteotomies of the humerus and radius and arthrodesis in the wrist or in the metacarpophalangeal joints of the thumb, depending on whether there is greater rigidity or age in the former cases or instability in the latter. PMID- 14533097 TI - [Advances in the molecular treatment of depression]. AB - AIMS: The objective of this study is to survey present and future antidepressant drug therapy based on the progress made in the field of biotechnology. DEVELOPMENT: The simplistic and mistaken view that one single system of neurotransmission is altered in depression and that there is, therefore, just one single treatment has changed. Molecular biology and Genetics have enabled us to determine other possible chemical alterations in the brain, beyond the sole participation of the monoaminergic modulation systems, which is the classical hypothesis. In this paper we describe the evidence for the relations between depression and the therapeutic effect the classical antidepressants have on: 1. The peptidergic system of the corticotropin-releasing hormone, cortisol and the functional state of its receptors; 2. Intracellular signalling systems such as cAMP on transcription factors like CREB and neurotrophins; 3. The immune system and cytosines; 4. Glutamate transmission; and 5. The neuropeptidergic system of substance P, neuroactive steroids and the neuroglia. This has allowed other biochemical hypotheses about depression and the possibility of new treatments to be put forward. CONCLUSIONS: We are still not certain about the exact cause or the processes that determine mental illnesses such as depression or how improvements are achieved with the antidepressants we currently have available. Nevertheless, biotechnology is expected to be a great help in advancing towards a better understanding of the interrelations between the nervous, immune and endocrine systems, with their intracellular cascades and final outcomes in genetic expression and protein function, in depression. This will enable more efficient, more selective and faster-acting drugs to be developed and, in the future and with the help of psychogenomics, even make it possible to produce tailor-made medication for each patient. PMID- 14533098 TI - [Alcoholism-related genetic alteration]. AB - AIMS: A number of studies about human genetics have shown how inheritance is involved in a predisposition towards alcohol abuse or towards developing alcoholism. Similarly, molecular genetics researchers have drawn attention to the importance of the enzymatic systems that metabolize ethanol and acetaldehyde in the study of alcoholism. These systems are essentially alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH), although the roles played by cytochrome P450 2E1 and catalase are notable. DEVELOPMENT: The existence of a functional polymorphism in some of the genes that codify for these enzymes could generate an alteration in the acetaldehyde synthesis interval or lower the extent to which it is later oxidized. A selection of this genetic polymorphism might act in certain human populations, such as the oriental population, as a factor that protects against alcohol abuse and the damage and injuries associated with this drug. CONCLUSION: Subjects with a sensitivity to alcohol that is genetically controlled by the abnormality contained in their allele for ALDH2*2 would present some kind of protection against excessive consumption of alcohol. Moreover, subjects with a heterozygotic genotype of ALDH2*2 (ALDH2*1/2*2) would present a greater risk of developing injuries and damage in the organs related to alcohol abuse as compared to subjects who present a homozygotic genotype of ALDH2*1/2*1. PMID- 14533099 TI - [Diseases of the peripheral and visual nervous system during infection with human immunodeficiency virus]. AB - INTRODUCTION: Infection with human immunodeficiency virus (HIV) is often accompanied by neurological complications. One of these includes disorders affecting the peripheral and visual nervous system, especially during the acquired immunodeficiency syndrome (AIDS) stage. DEVELOPMENT: The peripheral neuropathies associated with infection by HIV are an assorted group of disorders, which include acute or chronic inflammatory demyelinating polyneuropathy, multiple mononeuropathy and neuropathies related to the herpes zoster virus or cytomegalovirus. The most common and clinically important of the neuropathies is painful distal sensory polyneuropathy (DSP). The most severely affected cranial nerves are V and VII. The isolation of HIV from the affected nerves suggests a direct role, but an immune mechanism is also possible. Although cytomegalovirus may be associated with a variety of peripheral nerve syndromes, its clinical presentation as a primary demyelinating polyneuropathy is unusual. CONCLUSIONS: DSP and antiretroviral toxic neuropathy are the most common HIV-associated neuropathies. Both HIV infection, by itself, and the neurotoxicity of certain drugs in tritherapy contribute to the development of painful peripheral sensory neuropathy. In researching into the cause of HIV-associated neuropathy further studies are needed to determine the relative roles played by the viral infection and the activation of the immunological factors that contribute to the pathogenesis of the damage done in axons, the dorsal root ganglion and in the sensory pathways in the spinal cord. PMID- 14533100 TI - [The possible role of acetaldehyde in the brain damage caused by the chronic consumption of alcohol]. AB - AIMS: The purpose of this study is to collect and evaluate the experimental evidence suggesting that acetaldehyde, the first oxidative metabolite of ethyl alcohol (ethanol), plays a mediating role in the brain damage associated with the chronic consumption of this substance. DEVELOPMENT: Although the brain damage associated with the chronic consumption of ethanol is multifactorial and, possibly, dependent on the numerous actions of this substance on the central nervous system (CNS), there is data to suggest that the oxidative metabolism of ethanol and resulting substances are involved in the aetiology of such processes. Similarly, the generation of free radicals and the formation of adducts between the products of this metabolism and substrates contained in the CNS could be the main mediators in such pathological processes. This idea is supported by the fact that the adducts derived from the metabolism of ethanol are formed in the same areas of the brain as those which present structural and functional disorders in chronic consumers of alcohol. CONCLUSIONS: There are currently different experimental findings that appear to support the proposal that the substances resulting from the metabolism of ethanol can be important mediators in the brain damage associated with the chronic consumption of alcohol. Although more research is required, this theoretical proposal is very interesting, not just because of its relative novelty, but also because it is based on the same principles put forward to explain the toxic effects of alcohol consumption on organs and tissues. PMID- 14533101 TI - [The neurobiology of sleep: Cajal and present-day neuroscience]. AB - AIMS: We briefly describe the most significant findings obtained recently concerning the sleep-waking cycle in comparison to the studies conducted by Cajal on the same subject. DEVELOPMENT: This paper includes a short biographical sketch of Santiago Ramon y Cajal, with special emphasis on his importance within the framework of neuroscience. Cajal represents the decisive turning point in neurobiological studies, with the discovery of the synapse and his law of dynamic polarization. We conduct a short survey of the current knowledge about the phases of sleep and oneiric phenomena, based on their anatomo-physiological foundations. We present a summary of the history of the subject and analyze the contributions made by Cajal to this field, i.e. his study of the associative cortices, which are essential in memory processes and related to the mechanisms governing the sleep-waking cycle. For Cajal the fine anatomy of the thalamus must be considered in relation to the specificity of its connections an idea that is still completely valid today. He did not observe any projections of the thalamic reticular nucleus towards the cerebral cortex, a fact that has been corroborated using modern-day techniques. He spoke of the involvement of neuroglia in the attentional and sleep processes, which is so, although not quite in the way Cajal thought. He considered the production of dreams to be based on intimate neural mechanisms, which is still so. He also studied other brain structures related with the regulation of the sleep waking cycle, although avoiding any specific mention of the mechanisms controlling such a cycle. Furthermore, he conducted self-observation studies with a high degree of insight. CONCLUSIONS: Cajal studied the phenomena of attention and sleep in an objective manner and contributed a number of significant interpretations, some of which are now somewhat outdated while others are still wholly valid today. PMID- 14533102 TI - [Magnetic resonance findings in eclampsia]. PMID- 14533103 TI - [Scant knowledge about cerebrovascular disease in a sample of teenagers]. PMID- 14533104 TI - [Evaluation of hemodynamic parameters by transcranial Doppler in patients with leukoaraiosis]. AB - INTRODUCTION: Leukoaraiosis is commonly found in neuroimaging in ancient people. The pathogenic theory that most suitably explains its origin is the vascular one, mainly linked to cerebral hemodynamic abnormalities. The techniques most frequently used in cerebral hemodynamic evaluation (PET and SPECT) are expensive and not widespread. Transcranial Doppler instead is cheaper and much more widespread. OBJECTIVE: Our aim has been to show whether transcranial Doppler is a useful tool for cerebral hemodynamic evaluation in leukoaraiosis. PATIENTS AND METHODS: We have prospectively included 116 patients aged 60-90 who came to the Neurology department with unspecific complaints such as dizziness or mild headache. Patients with recent history of stroke or moderate to severe cognitive impairment were excluded. RESULTS: Mean age was 74.4 +/- 6.3 years old. The prevalence of leukoaraiosis was 68.7%. Leukoaraiosis was significantly correlated with older age, lacunar infarctions, past history of stroke, cognitive impairment and to lower systolic and diastolic velocities in middle cerebral artery (MCA) and higher pulsatility index. Multivariate analysis only retained the lower diastolic velocity in MCA in the model. CONCLUSIONS: Transcranial Doppler can detect hemodynamic abnormalities in patients with leukoaraiosis and is therefore a very useful technique for the evaluation of this entity. PMID- 14533105 TI - [Retrograde suction decompression technique for paraclinoidal aneurysms]. AB - INTRODUCTION: Paraclinoidal carotid artery aneurysms pose technical surgical problems with regard to acquisition of the proximal control and safe intracranial exposure due to brisk retrograde flow through the ophthalmic artery and cavernous branches. Simple trapping of the aneurysm by cervical internal carotid artery clamping and intracranial clipping may not adequately soften the lesion. PATIENTS AND METHODS: Fifteen patients with giant and complexes large paraclinoidal aneurysms were operated on. We utilized microneurosurgical techniques, specialized neuroanesthesia, mild hypothermia, barbiturical brain protection, electroencephalographic monitoring and retrograde suction decompression technique described by Batjer et al. Patients were followed at Intensive Care Unit and were evaluated three months later. RESULTS: . 74% were female and there was a 14% of multiplicity. There was not intraoperative aneurysmal rupture and all aneurysms were clipped. We obtained 67% of good recovery, 27% of partial recovery, and no patient died. CONCLUSION: Retrograde suction decompression technique was useful for surgical treatment of paraclinoidal aneurysms. We introduced three technical modifications to the original procedure: intraoperative mild hypothermia, heparin utilization for endovascular procedure and intraoperative surgical evaluation of the vascular anatomical wall to select optimal area for angiocatheter insertion and temporary clipping. Modifications improve results. PMID- 14533106 TI - [Delayed-type subacute measles encephalitis]. AB - INTRODUCTION: We analyze the clinical, neurological, EEG, neuroradiological features and evolution of two patients with subacute measles encephalitis. CASE REPORTS: The patients, aged five years and eleven months respectively showed an acute, progressive neurological compromise and deterioration of consciousness, epilepsia partialis continua and progressive damage on neuroimaging, with a history of measles in the first case and exposure to the virus in the second. The first patient had Hodgkin's disease and the other had a familial C4 deficit disorder. Fundoscopic examination showed lesions on the retina. The EEG showed unilateral slow waves and spikes. Brain CT and MRI revealed progressive cerebral atrophy and a unilateral corticosubcortical lesion. Measles antibodies in CSF were found in the first child and oligoclonal bands in the second. Our first patient died after three months and the second has a severe neurological damage. CONCLUSION: In immunocompromised patients with the exposure to a history of measles, acute neurological compromised and deterioration of consciousness, epilepsia partialis continua and progressive damage on neuroimaging, subacute measles encephalitis should be considered. PMID- 14533107 TI - [The efficiency of carbamazepine in a case of post-streptococcal hemichorea]. AB - INTRODUCTION: Chorea is an infrequent disorder at the paediatric age which has a number of both hereditary and acquired causes. Post-streptococcal or Sydenham's chorea (SC) is still the main cause of chorea in children, in spite of the drop in prevalence of rheumatic fever in the last few years. SC is a benign, self limiting disorder, but may last for several months and can be highly disabling. Several different types of symptomatic treatment have been proposed, for example corticoids, haloperidol, valproic acid, and carbamazepine. In each case, both the speed with which the clinical improvement is brought about, and the degree to which they are tolerated and the absence of side effects must be evaluated. CASE REPORT: We present a new case of SC that had been developing for three months. Carbamazepine was effective from the tenth day onwards and total remission of the symptoms was achieved in six weeks. Total follow-up time was nine months, and in this time no relapses or side effects were observed. CONCLUSIONS: This contribution offers new evidence supporting carbamazepine as another first choice medication in the treatment of this type of chorea. PMID- 14533110 TI - [Malformations of cortical development and their clinical repercussions in a series of 144 cases]. AB - OBJECTIVE: To show the disorders of the brain cortical development and the possible origin in base to a large series studied in a Pediatric Neurology service. PATIENTS AND METHODS: A series of 144 children with ages ranging between newborn and 12 years was studied from the clinic, image (MR, 3DMR) and evolutive point of views. RESULTS: The diagnosis was: polymicrogyria in 61 cases, lissencephaly in 22, eschizencephaly in 16, heterotopia in 16, cortical dysplasia in 9, hemimegalencephaly in 8, cobblestone in 7, sublobar dysplasia in 3, and 'double cortex' in 2. Mental retardation, motor disorders and epilepsy were the most important anomalies. CONCLUSIONS: Actually, the image is the most important study to make the diagnosis of every type of cerebral malformation. However, to know the specific gene that origin every disorder seems to be the most important thing to make the classification of every malformative type and the possible prevention of this pathology. PMID- 14533111 TI - [West's syndrome. Analysis, aetiological factors and therapeutic options]. AB - West's syndrome (WS), which is also known as infantile myoclonic encephalopathy with hypsarrhythmia, is one of the generalized epileptic syndromes with a cryptogenic or symptomatic origin. It is an age-dependent epileptic syndrome. The latest neuroimaging techniques have enabled us to gain a better understanding of its physiopathology and to identify new aetiological factors responsible for the clinical symptoms. WS can be due to a number of aetiologies, the most notable of which are congenital errors of metabolism. The incidence of cases due to phenylketonuria or hypoglycaemia is currently diminishing, yet, there is a rise in the number of new metabolic diseases that are responsible for the symptoms of WS. These include the carbohydrate-deficient glycoprotein syndromes or biotinidase deficiency. In all cases, and especially so in those that are idiopathic, it is wise to conduct exhaustive aetiological studies, since on some occasions metabolic diseases will be shown to be responsible, and this will then modify the prognosis, therapy and genetic counselling. It is important to have a protocol for both study and therapy available for this syndrome. The therapeutic options available can be implemented after ruling out a neurometabolic disease as being responsible for the syndrome and quickly beginning treatment with vigabatrine, sodium valproate plus pyridoxine, ACTH or hydrocortisone. If there is no response then topiramate can be used. Other therapeutic options, such as the use of zonisamide, a ketogenic diet or even surgical treatment, are also analyzed. PMID- 14533112 TI - [Attention deficit hyperactivity syndrome]. AB - INTRODUCTION: Attention deficit/hyperactivity disorder (ADHD) is an early onset childhood condition that is estimated to occur in 7-10 % of school age children. DEVELOPMENT: There are two specific syndromes: a) Attention deficit disorder, without hyperactivity; b) A more complex syndrome, with hyperactivity, with traits of impulsivity, accompanied by attention deficit disorder. The etiology of this syndrome is still unclear, but there have been some hypothesis about hypoperfused areas in the frontal lobe, in some patients where PET has been available. There have been also some response to stimulants; which leads to the conclusion that there is some relation with neurotransmitters like noradrenaline and dopamine, as a determinant factor in the origin of this process. Among 80% of children with this diagnosis, respond positively to the use of psychostimulants. Although, there are some cases that respond poorly, which are evaluated in this review. CONCLUSION: This article also highlights the importance of multidisciplinary assessment, in the diagnosis and treatment of these cases. It gives a clinical characterization of the syndrome, following DSM IV guidelines. Finally, it addresses, the use of non stimulant drugs, that can be helpful in the treatment of this disorder. PMID- 14533113 TI - [Hyperbaric oxygen treatment for children with cerebral palsy]. AB - AIMS: Demand from parents has made hyperbaric oxygenation (HO) inhalation the most popular and rapid growing therapy for children with cerebral palsy (CP). To review peer reviewed articles of HO in children with CP to determine its efficacy and risks, literature was searched on-line using PubMed indexed for MEDLINE (1996 2003) for articles under CP and HO headings. METHOD: We found 16 references: 5 articles (1 uncontrolled pilot study, 2 from a single controlled study, 1 case report of complications and 1 revision) and 11 letters to the editor. The control study showed significant improvements in the middle, at the end and 3 months after 40 treatments with OH (O2=100%/1.75 AA) and placebo (O2=21%/1.3 AA) in the gross motor function measure, (2.9% vs 3%), self-control, auditory attention and visual working memory. There were no significant differences between the groups. Side effects included barometric otitis media (48.2% and 22.2% in the OH and placebo groups). The authors and the Advisory Scientific Committee of the American Academy of Cerebral Palsy and Developmental Medicine agreed that the positive results in both groups were due to a participation effect. The Southern Africa Undersea and Hyperbaric Association discouraged the ongoing, widespread, and informal use of HO for children with CP in South Africa based on the results of this randomized controlled study. CONCLUSION: There are no scientific evidences for the use of HO in children with CP. Risks include barometric otitis media. PMID- 14533114 TI - [Headaches in children and adolescents. An etiopathogenic classification]. AB - Headache is one of the most frequent reasons for presenting to Neuropaediatric units. Although in most cases the patient is not suffering from any serious disease, the parents and even the paediatrician do tend to fear that this is indeed the case. Evaluating a patient with headache requires a detailed interview, as well as a careful clinico-neurological examination. From these elements it is basically possible to sort headaches into four large groups: acute, acute-recurrent, chronic non-progressive and chronic progressive. Acute headaches are an emergency and it is therefore necessary to have an algorithm available which can be used regardless of the degree of complexity of the clinical environment in which the patient is being treated. Invasive studies such as a spinal tap and/or neuroradiological studies are often needed. The International Headache Society criteria have been adapted to the juvenile population for the diagnosis of patients suffering from acute-recurrent headaches (migraine). Chronic non-progressive headaches are a group in which prevalence increases with age and is the most frequently observed form of headache in adolescents. Emotional factors, situations involving family and/or social conflict, either alone or in combination, are usually predisposing or triggering factors in this type of headache. Chronic progressive headache is a consequence of some organic disorder. In the search for and treatment of headache we must not forget to calm the pain experienced by the patient. PMID- 14533115 TI - [The treatment of children with difficult to control epilepsy]. AB - A survey is conducted of the way difficult-to-control epileptic seizures are currently managed in paediatric practice. We also highlight the alternative means of therapy available, such as epilepsy surgery, a ketogenic diet, the use of hormones, steroids, gamma globulin and the stimulation of the vagal nerve, together with their indications, their efficiency in the different types of epilepsy and their contraindications. Mention is also made of the new antiepileptic drugs that have appeared since the nineties, as well as the reappearance of others that had fallen into disuse. PMID- 14533117 TI - [Parkinson's disease as seen by a patient with this disease]. AB - INTRODUCTION: The idea of writing this paper stemmed from the interest shown by Dr Francisco Javier Jimenez-Jimenez in giving a firsthand description of the experiences of a patient suffering from Parkinson's disease, as a complement to the scientific view. The author is not a member of the medical profession. DEVELOPMENT: This study is based on the analysis of the disease as seen by the person actually suffering from it. From this biography we analyze the possible causes that could have exerted an influence on the development of the disease, and the physical causes are also outlined. We analyze the drugs that are administered as therapy and we establish the relations that exist between the symptoms presented by the author and the possible deterioration of the governing centres of the brain which should control them but, due to the degeneration brought about by the disease, do not. The effects produced by the medication are studied and we also describe how help can be obtained from the outside by employing other types of medicine that are not strictly allopathic. In the final part of the paper the relations between the patient and the people around them are discussed. CONCLUSIONS: It becomes clear that there is a void in our knowledge about this disease due to our not knowing what exactly causes it. This means that therapies are directed towards palliating the symptoms rather than fighting the causes. Something that must be highlighted is the important role played by the patient's will to fight against the disease. PMID- 14533118 TI - SARS and psychogeriatrics: perspective and lessons from Hong Kong. PMID- 14533119 TI - Consumption of psychotropic medication in the elderly: a re-evaluation of its effect on cognitive performance. AB - BACKGROUND: There have been few general population studies of the effects of psychotropic treatment on cognitive functioning in the elderly. Current evidence based on studies with numerous procedural shortenings supports the notion of the detrimental effect. OBJECTIVES: To examine changes in a wide range of specific cognitive abilities across time in a general population sample in order to establish a relationship between psychotropic drug use and cognitive performance, and to estimate to what extent such cognitive changes may be attributable to psychotropic use or other factors, notably age and co-morbidity. METHOD: We analysed the data from the Eugeria longitudinal study of cerebral ageing. Three hundred and seventy two subjects (263 female and 109 male) were visited at their place of residence and given a computerized cognitive examination. Depressive symptomatology and depressive episodes were defined according to ICD-9 criterias and medication use were established. Four categories of psychotropic consumers was differentiated. Using a logistic regression model, comparisons were made between consumers and non-consumers. RESULTS: A significant positive effect in chronic consumers was found on tests of secondary memory (delayed verbal recall: Odds Ratio (OR)=1.22; 95% Confidence Intervals (CI) [1.04-1.43]; p=0.013) and this effect is principally attributable to antidepressants with significant effects being shown for both verbal (OR=1.59; 95%CI [1.18-2.14]; p=0.002) and visual recall (OR=1.51; 95%CI [1.05-2.16]; p=0.025). No effect is found for benzodiazepines. CONCLUSIONS: Contrary to the common belief that psychotropic drug use has a detrimental effect on cognitive function of elderly people, even long term use is seen to be benign. We attest to the positive effects of antidepressant therapy on secondary memory. PMID- 14533120 TI - Clinical predictors of response to Acetyl Cholinesterase Inhibitors: experience from routine clinical use in Newcastle. AB - BACKGROUND: Acetyl Cholinesterase Inhibitors (AChEIs) have been in clinical use for the past five years in the UK for the symptomatic treatment of Alzheimer's disease (AD). There are few data on the patterns and predictors of response to AChEI therapy in routine clinical practice. We therefore investigated clinical variables that may distinguish between AChEI responders and non-responders. METHODS: A retrospective sample of 160 consecutive patients with dementia who were treated on clinical grounds with an AChEI was studied. Treatment response was defined in two ways: (a) A clinical response was achieved when there was no deterioration or there was an improvement on a global clinical assessment (CGI) and (b) a Mini-Mental-State-Examination (MMSE) response when there was an improvement of 2 or more MMSE points. RESULTS: A total of 62 (45%) patients achieved an MMSE response. A diagnosis of dementia with Lewy Bodies (DLB) and Parkinson's disease+Dementia (PDD) was associated with a MMSE response, as were hallucinations, and lower MMSE scores at baseline. 125 (78%) patients achieved a CGI response for which there were no clinical predictors. CONCLUSIONS: Severity of illness, a diagnosis of DLB and PDD, and presence of hallucinations at baseline were predictive of a MMSE response. Non-AD dementia and severe dementia responded equally well to AChEI treatment and results of further randomised, placebo-controlled studies are needed to clarify the role of AChEI in the treatment of these disorders. PMID- 14533121 TI - Does integration really make a difference? A comparison of old age psychiatry services in England and Northern Ireland. AB - OBJECTIVE: This paper seeks to address whether integrated structures are associated with more integrated forms of service. Northern Ireland has one of the most structurally integrated and comprehensive models of health and personal social services in Europe. Social and health services are jointly administered and this arrangement should, in theory, promote collaborative working and interdisciplinary arrangements. DESIGN: The study employed a cross-sectional survey of consultants in old age psychiatry in England and Northern Ireland. Potential respondents were sourced from the UK Royal College of Psychiatrists membership list and locally collected information. MEASURES: A self-administered postal questionnaire. Along with general service arrangements, the domains measured reflect core policy issues for older people's services. Under particular scrutiny in this study were the degree of integration of health and social service provision, as well as inter-professional team working. RESULTS: The integrated health and social care services in Northern Ireland do appear to provide more integrated patterns of working, primarily in managerial arrangements and in the location of staff. There was no evidence of the impact of integration on practice in areas such as: assessment, referral and medical screening. The factors found to be associated with greater integration of health and social care in the prediction model fell into three categories: provision of specialist services; provision of outreach activities; and shared policies by which the whole team worked. CONCLUSIONS: Health and personal social services in Northern Ireland have a distinct advantage over their counterparts in comparable areas of England. The results indicate that integrated structures in old age psychiatry services are associated more with integrated management systems and less with integrated practice-related activities. Further research is required on the effectiveness and cost effectiveness of integrating services in general. It is important that future intervention studies systematically measure the component parts, nature and extent of integration and their individual and joint contribution to the effectiveness and efficacy of services. PMID- 14533122 TI - The efficacy and safety of ECT in depressed older adults: a literature review. AB - BACKGROUND: Although little doubt exists among practising clinicians in old age psychiatry about the efficacy and safety of ECT in depression, opinions about acceptability differ widely. The objectives of this review were to determine the efficacy and safety of ECT based on both randomised and non-randomised evidence in elderly with a major depressive disorder. METHODS: Randomised and non randomised studies on efficacy and safety of ECT in elderly with and without concomitant disorders such as cerebrovascular disorders, Alzheimer's dementia, vascular dementia and Parkinson's disease were selected. Literature was systematically searched in a number of electronic databases. RESULTS: Although 121 studies were included in the review process, only four provided randomised evidence. No negative studies with respect to efficacy were found. ECT is effective in the acute treatment of late life depression. ECT is generally safe, although a number of serious complications possibly related to ECT have been described. Most of the objectives of this review could not be answered or refuted with certainty, because firm randomised evidence on the efficacy and safety of ECT in the depressed elderly is missing. CONCLUSIONS: ECT is effective in the acute treatment of late life depression and is generally safe. Important questions such as the relative efficacy of ECT over antidepressants, the long term efficacy of ECT, morbidity and mortality related to ECT, cost-effectiveness and the efficacy of ECT in subgroups of patients cannot be answered and need to be studied further. PMID- 14533123 TI - The impact of religious practice and religious coping on geriatric depression. AB - OBJECTIVE: Both religiousness and social support have been shown to influence depression outcome, yet some researchers have theorized that religiousness largely reflects social support. We set out to determine the relationship of religiousness with depression outcome after considering clinical factors. METHODS: Elderly patients (n=114) in the MHCRC for the Study of Depression in Late Life while undergoing treatment using a standardized algorithm were examined. Patients completed measures of public and religious practice, a modified version of Pargament's RCOPE to measure religious coping, and subjective and instrument social support measures. A geriatric psychiatrist completed the Montgomery-Asberg Depression Rating Scale (MADRS) at baseline and six months. RESULTS: Both positive and negative religious coping were related to MADRS scores in treated individuals, and positive coping was related to MADRS six months later, independent of social support measures, demographic, and clinical measures (e.g. use of electro-convulsive therapy, number of depressed episodes). Public religious practice, but not private religious practice was independently related to MADRS scores at the time of completion of the religiousness measures. Religious coping was related to social support, but was independently related to depression outcome. CONCLUSIONS: Clinicians caring for older depressives should consider inquiring about spirituality and religious coping as a way of improving depressive outcomes. PMID- 14533124 TI - Diagnosis-seeking at subspecialty memory clinics: trigger events. AB - BACKGROUND: Research has shown that dementia often goes unrecognized, and diagnostic assessment is often further delayed. Understanding families' decision to seek care at memory clinics is relevant to efforts to facilitate early diagnosis. OBJECTIVE: To examine the population seeking care at two memory clinics and the triggers causing caregivers to seek diagnostic assessment for a family member. METHOD: We surveyed a consecutive sample of caregivers who accompanied a patient to an assessment at two university memory disorders clinics. Caregivers (n=416) described events that led them to seek a memory assessment for the patient, as well as who first suggested an assessment and diagnosis received. RESULTS: Changes in the patient (cognitive, personality/behavioral, physical, or unspecified) accounted for 81% of 903 trigger events reported. Nearly half of the caregivers noting specific patient changes recorded some combination, rather than cognitive changes alone. Of the 338 respondents who noted a change in the patient as a trigger, 85% specified at least one cognitive change, while 40% specified at least one personality/behavioral change. Memory loss was most frequent trigger reported, followed by disorientation and recommendations (lay or professional). Caregivers themselves and non-specialist physicians were the most frequent sources of recommendations noted by all respondents. CONCLUSION: A broad range of trigger events, beyond cognitive or symptomatic changes, caused caregivers to seek diagnosis at a memory clinic. Awareness of triggers significant to families may help physicians reduce the number and severity of events needed to convince caregivers a memory assessment is indicated. PMID- 14533125 TI - Validation of a short cognitive tool for the screening of dementia in elderly people with low educational level. AB - AIM: To validate the 'Prueba Cognitiva de Leganes' (PCL) as a screening tool for cognitive impairment in elderly people with little formal education. METHODS: The PCL is a simple cognitive test with 32 items that includes two scores of orientation and memory and a global score of 0-32 points. It was applied to a population sample of 527 elderly people over 70 with low educational level, who were independently diagnosed by consensus between two neurologists as having normal cognitive function, age associated cognitive decline (AACD, IPA-OMS criteria) or dementia (DSM-IV criteria). Individuals with severe visual or hearing defects and those who rejected the exam were excluded from the study. The PCL was validated in a sample of 375 individuals: 300 normal, 42 with AACD and 33 with dementia. The sensitivity, specificity, accuracy and likelihood ratios, as well as the ROC curves for dementia and for AACD-dementia, were calculated. The confounding effect of sociodemographic variables was assessed by logistic regression analysis and convergent validity by partial correlations of the PCL with other cognitive tests. Inter-rater reliability was evaluated with the intraclass correlation coefficient. RESULTS: The PCL identified dementia (cut-off < or =22) and AACD-dementia (cut-off < or =26), with the following diagnostic parameters, respectively: sensitivity 93.9%-80%, specificity 94.7%-84.3%, positive likelihood ratio 17.8-5.1, negative likelihood ratio 0.06-0.24, and accuracy 94.6%-83.4%. The areas under the ROC curve were 0.985 (95% Confidence Intervals (CI) 0.967-0.995) and 0.904 (95% CI: 0.870-0.932) respectively. The intraclass correlation coefficient was 0.79 (0.74-0.83). CONCLUSION: The PCL is a simple instrument, which is both valid and reliable, for the screening of dementia in population samples of individuals with low educational level. This instrument could be useful in primary health care. PMID- 14533126 TI - Cognitive, psychiatric and motor response to galantamine in Parkinson's disease with dementia. AB - BACKGROUND: Cholinesterase inhibitors with additional nicotinic activity, such as galantamine, may be useful in PD patients with dementia (PDD) since stimulation of nicotinic receptors may prevent the down-regulation that is likely to accompany cholinesterase inhibition and facilitate dopamine release in the striatum. METHODS: Sixteen PDD patients (six female) with onset of cognitive impairment after at least one year with parkinsonism participated in this open label trial of galantamine. Cognitive, psychiatric, and motor symptoms were assessed before and after 8 weeks of treatment with galantamine using unstructured clinical assessment as well as rating scales including the Mini Mental State Examination (MMSE), clock drawing test, verbal fluency and selected items from the Neuropsychiatric Inventory (NPI). RESULTS: Age (mean, SD) was 75.6 (5.2) years, duration of PD 13.4 (5.9), duration of dementia 2.1 (1.7) years, Hoehn and Yahr score was 3.8 (0.8) and baseline MMSE score was 17.7 (6.7). Side effects caused discontinuation in three patients, but were rare and mild in the remaining 13. Improvement of global mental symptoms was noted in eight patients, whereas worsening was reported in four. Hallucinations improved in seven of the nine patients with hallucinations before treatment. Parkinsonism improved in six patients, but a mild worsening of tremor was noted in three. Clock-drawing improved (p=0.016), and trends towards improvement on MMSE (p=0.09) and verbal fluency (p=0.16) were found. CONCLUSIONS: Although controlled trials are needed, the findings suggest that galantamine is useful in patients with PDD. PMID- 14533127 TI - The effects of galantamine treatment on caregiver time in Alzheimer's disease. AB - AIM: The aim of the study was to determine whether the clinical benefits of galantamine for patients with Alzheimer's disease lead to benefits for caregivers. METHODS: Data were pooled from two concurrent, multi-centre, randomized, double-blind, placebo-controlled, 6-month trials. Time caregivers spent assisting with activities of daily living (ADL) and time patients could be left unsupervised each day were assessed using the Allocation of Caregiver Time Survey. In total, 825 patients with mild-to-moderate Alzheimer's disease were included. RESULTS: At endpoint, caregivers of galantamine-treated patients were more likely to report reductions (41% vs 37%), maintenance (19% vs 14%) or smaller increases (26% vs 34% reporting an increase >30 minutes) in time assisting with ADL compared with the placebo group (p=0.026; Wilcoxon rank-sum test). The mean daily time difference was 32 minutes (p=0.011). Among patients with moderate Alzheimer's disease, caregivers of galantamine-treated patients were even more likely to report reductions (46% vs 37%), maintenance (15% vs 6%) or smaller increases (25% vs 42% for increases >30 min) vs placebo (p=0.004), with a mean daily time saving of 53 minutes (p=0.021). Caregivers of galantamine treated patients were more likely to report increases (22% vs 18%), maintenance (45% vs 43%) or smaller reductions (30% vs 37% for reductions >30 minutes) in time the patient could be left unsupervised compared with placebo (p=0.027). Mean daily time saving was 27 minutes. Among patients with moderate Alzheimer's disease, the treatment effect was greater (p=0.029), with caregivers in the galantamine group reporting the change in time left unsupervised as 68 minutes longer each day than caregivers of patients receiving placebo. CONCLUSION: The clinical benefits of galantamine for patients with Alzheimer's disease are also associated with benefits to caregiving. PMID- 14533128 TI - Anxiety disorder and accompanying subjective memory loss in the elderly as a predictor of future cognitive decline. AB - OBJECTIVE: The aim of the study was to empirically investigate the hypothesis that anxiety in the elderly, secondary to loss of memory, predicts future cognitive decline. METHOD: The participants were 137 elderly subjects with no depression or cognitive impairment from a community geriatric assessment unit, 45% with anxiety. In addition to demographic characteristics, cognitive status was assessed using the Mini Mental State Examination; depression was assessed by Tucker's short Interviewer-Assisted Depression Rating Scale; anxiety by Sinoff's Short Anxiety Screening Test and Activities of Daily Living function by Shah's modified Barthel's Index. RESULTS: At follow-up 37 persons had dropped out, leaving 100 participants for final analysis. Mean re-examination time was 3.2 years with no group differences. The mean MMSE and modified Barthel scores decreased significantly more in those with anxiety. A relative risk of 3.96 for developing future cognitive impairment was found. Regression analysis showed that only anxiety was a significant predictor of cognitive decline. By path analysis, a more parsimonious model showed anxiety to have both a direct and an indirect effect on predicting future cognitive decline, and that the effect of loss of memory on cognitive decline was via anxiety. CONCLUSIONS: Anxiety is inter related and inseparable with loss of memory and its presence is a strong predictor for future cognitive decline, directly or indirectly via depression. It appears that loss of memory is the initial problem with consequent development of anxiety. Therefore, anxiety, like depression, is probably an early predictor of future cognitive decline and even possible future cognitive impairment. PMID- 14533129 TI - Does personality disorder influence the likelihood of in-patient admission in late-life depression? PMID- 14533130 TI - Post-marketing studies: the Italian CRONOS project. PMID- 14533132 TI - Acute, sub-acute and long-term subjective consequences of 'ecstasy' (MDMA) consumption in 430 regular users. AB - This study examined the reported psychological effects of different patterns of MDMA use in men and women, and how they are modified by use of other psychoactive substances. A semi-structured interview was conducted with 466 regular MDMA users, exploring the perceived acute, sub-acute and long-term subjective effects of this drug. Factor analysis established three main categories of acute effects of MDMA: (i) positive and (ii) negative effects on mental health, and (iii) physical effects. In terms of subacute effects, 83% of participants reported experiencing low mood and 80% reported impaired concentration between ecstasy taking sessions. Factors affecting these effects included age, gender, extent of MDMA use and concomitant use of cocaine or amphetamine. The long-term effects most frequently reported included the development of tolerance to MDMA (59%), impaired ability to concentrate (38%), depression (37%) and 'feeling more open towards people' (31%). In terms of what might persuade users to stop using MDMA, their most prominent concern was the drug's long-term effects on mental health. PMID- 14533133 TI - Death rates from ecstasy (MDMA, MDA) and polydrug use in England and Wales 1996 2002. AB - The present study reports on all deaths related to taking ecstasy (alone, or in a polydrug combination) occurring in England and Wales in the time frame August 1996-April 2002. Data presented here are based on all information recorded in the National Programme on Substance Abuse Deaths (np-SAD) database. The np-SAD regularly receives all information on drug related deaths in addicts and non addicts from coroners. A total of 202 ecstasy-related fatalities occurred in the chosen time-frame, showing a steady increase in the number of deaths each year. The ratio male:female was 4:1 and 3 of 4 victims were younger than 29. In 17% of cases ecstasy was the sole drug implicated in death and in the remaining cases a number of other drugs (mostly alcohol, cocaine, amphetamines and opiates) have been found. According to toxicology results, MDMA accounted for 86% of cases and MDA for 13% of cases; single deaths were associated with MDEA and PMA. This is the largest sample of ecstasy related deaths so far; possible explanations are given for the observed steady increase in ecstasy-related deaths and a tentative 'rationale' for this polypharmacy combination is then proposed. PMID- 14533134 TI - Saccade and cognitive impairment associated with kava intoxication. AB - Kava is an extract from the Piper methysticum Forst. f. plant that has social and spiritual importance in Pacific islands societies. Herbal remedies that contain kava are used for the psychiatric treatment of anxiety and insomnia. Laboratory studies have found only subtle, if any, changes on cognitive or motor functions from the acute effects of consuming small clinical doses of kava products. Intoxication from recreational doses of kava has not been studied. The performance of individuals intoxicated from drinking kava (n=11) was compared with a control group (n=17) using saccade and cognitive tests. On average, intoxicated individuals had consumed 205 g of kava powder each (approximately 150 times clinical doses) in a group session that went for 14.4 h and ended 8 h prior to testing. Intoxicated kava drinkers showed ataxia, tremors, sedation, blepharospasm and elevated liver enzymes (GGT and ALP), together with saccadic dysmetria, saccadic slowing and reduced accuracy performing a visual search task that only became evident as the task complexity increased. Kava intoxication is characterized by specific abnormalities of movement coordination and visual attention but normal performance of complex cognitive functions. Saccade abnormalities suggest disruption of cerebellar and GABAergic functions. PMID- 14533135 TI - Preliminary investigation of the association between psychotropic medication use and accidents, minor injuries and cognitive failures. AB - RATIONALE: Psychotropic medication may impair psychomotor and cognitive function, and so increase accident susceptibility. However, few studies examine any association in the framework of other demographic, health and occupational factors. OBJECTIVES: To examine the relationship between sleeping tablets and antidepressants, and accidents, minor injuries and cognitive failures. METHODS: Postal questionnaire study of 3111 individuals who participated in an earlier phase of the Bristol Stress and Health Study. RESULTS: Sleeping tablet use was associated with accidents, minor injuries and cognitive failures both at work and outside work. Antidepressant use was associated with accidents and cognitive failures both at work and outside work. Adjusting for health, demographic and occupational factors explained many of these associations, but sleeping tablet use was independently associated with accidents (both at work and outside work), and antidepressant use with cognitive failures (outside work). CONCLUSIONS: Antidepressant and sleeping tablet use were apparently associated with accidents, minor injuries and cognitive failures. Although most of the associations could be accounted for by other factors the results suggested links between sleeping tablet use and accidents, and between antidepressant use and cognitive failures. PMID- 14533136 TI - Stem-completion tasks (indirect, direct inclusion and exclusion) are differently affected by equipotent doses of lorazepam and flunitrazepam. AB - This study was designed to explore the effects on performance in stem-completion tasks of two benzodiazepines (BZ) in equipotent doses: lorazepam, a drug that atypically disrupts perceptual priming, and flunitrazepam, a compound with standard BZ effects. The study followed a placebo-controlled, double-blind, parallel-group design. Thirty-six young and healthy subjects carried out three completion tasks at theoretical peak-plasma concentrations of drugs: (a) indirect tasks, in which the subjects were instructed to complete stems with the first word that came to mind; (b) direct inclusion tasks/cued recall, in which the participants had to try to use words seen at study as completions; and (c) direct exclusion tasks, in which words seen at study were to be avoided. The PDP was applied to the results in the inclusion and exclusion tasks, to obtain indices of explicit/controlled (C) and implicit/automatic (A) memory. The C index was lowered by both BZs and A was equivalent in all treatments, confirming the general amnestic action of BZs. However, lorazepam led to decreases in completions in the indirect and inclusion tasks, while flunitrazepam impaired performance in the exclusion task. The qualitative differences between the drugs in their effects on performance suggest that these BZs may lead to differences in response bias. PMID- 14533137 TI - The relationship of changes in leptin, neuropeptide Y and reproductive hormones to antipsychotic induced weight gain. AB - OBJECTIVES: Weight gain is an important side effect of antipsychotic (AP) treatment. Weight is regulated by multiple systems, including leptin, neuropeptide Y (NPY) and gonadal steroids. The aim was to investigate whether AP induced weight gain was related to leptin and NPY abnormalities and whether these were associated with a disruption of gonadal steroid production. METHODS: Twenty two female patients with schizophrenia receiving standard AP treatment were studied over a 3-month period. Plasma leptin, NPY, gonadal steroids and their regulators were measured along with weight and BMI. RESULTS: Weight, leptin and testosterone levels increased over time. There were significant relationships between a change in oestrogen levels and both a change in NPY levels and a change in BMI. Change in BMI, weight and leptin all correlated strongly with a change in the testosterone/luteinizing hormone ratio. CONCLUSIONS: AP treatment results in increase in weight over time and this increase is accompanied by increased leptin levels. AP-induced weight gain is also associated with disruption of the hypothalamic-pituitary-gonadal axis. Altered regulation of NPY, either through abnormal leptin control or serotonin blockade, is a possible explanation for the effects of AP medication on both weight and gonadal steroid levels. PMID- 14533138 TI - Quetiapine as adjunctive treatment of a case of rapid-cycling bipolar disorder with comorbidity. PMID- 14533139 TI - Donepezil in a chronic drug user--a potential treatment? AB - The objective of the current study was to explore the potential cognitive benefits of an anticholinesterase inhibitor, donepezil, in a former chronic drug user. A neuropsychological test battery composed of the vocabulary and matrix reasoning subtests of the Wechsler adult intelligence scale-III, measures of everyday executive functioning (behavioural assessment of the dysexecutive syndrome [BADS]), and verbal learning and memory tasks (California verbal learning test-II; Rivermead behavioural memory test) was completed at baseline, at 3 months after introducing donepezil, and at 3 months after donepezil was discontinued. After donepezil treatment, substantial improvements were found on tasks of nonverbal fluid reasoning (i.e. matrix reasoning) and other executive functioning tests (i.e. BADS). At entry into the study, poor academic performance and subjective problems with memory and concentration were reported, particularly after amphetamine use (i.e. MDMA and crystal methamphetamine); after donepezil treatment, dramatic increases in memory, concentration and academic achievement were observed. The finding of improvements in tests of executive functioning and in academic performance in this case study, together with the minimal adverse side effects of donepezil, warrants the investigation of controlled studies of cholinergic enhancement in chronic amphetamine and other drug users. PMID- 14533140 TI - Induction of mania in depression by paroxetine. AB - INTRODUCTION: An investigation of the proportion of patients who have experienced mania with antidepressant treatment and their characteristics would seem to be of clinical use. AIMS: The purpose of this clinical study was to examine the predictors of induction of mania in depression patients as a result of paroxetine treatment. METHOD: A retrospective cohort analysis was carried out among depression patients treated in the Department of Psychiatry, Kawasaki Medical School Hospital, Kurashiki, Japan, in 2000 and 2001. Some 79 patients were identified who were receiving paroxetine to treat depression. A variety of clinical factors including gender, the type of depression, frequency of episodes, family history, age and daily dose were examined as possible predictors of induction of mania by paroxetine. RESULTS: Seven (8.86%) of the 79 paroxetine treated patients developed mania. A Cox proportional hazards analysis showed the type of depression and the history of family psychiatric illness to be independent predictive factors of the induction of mania by paroxetine treatment. CONCLUSIONS: The rate of mania induction of paroxetine is not substantially different from that found for conventional antidepressants. The type of depression and the history of family psychiatric illness may be considered as predictors of mania induction in depression patients taking paroxetine treatment. PMID- 14533141 TI - Effect of pindolol and milnacipran versus milnacipran and placebo on plasma prolactin and adrenocorticotrophic hormone in depressed subjects. AB - OBJECTIVE: Evidence that pindolol accelerates the action of antidepressants has been contradictory, and it is not clear why. The present study analyses the relationship between plasma prolactin (PRL) and ACTH levels and changes in relation to a milnacipran and pindolol combination versus milnacipran plus placebo. METHOD: Eighty depressed patients agreed to take part in a double-blind randomized trial of milnacipran plus pindolol or placebo. Fifty-eight of them agreed to also take measures of ACTH and PRL levels. ACTH and PRL plasma levels were estimated on days 0 and 42 of the 6-week study. Age, gender and time of blood collection were recorded for each individual. The Montgomery-Asberg depression rating scale (MADRS) was used to measure the response to treatment. The patients were grouped into those with higher versus lower basal ACTH levels using the median of the sample (25 ng/l). RESULTS: There were statistical differences in MADRS scores between the treatment groups on day 42. There were correlations between PRL levels on days 0 and 42; age and PRL levels on day 0; time of the PRL sample and the PRL levels on day 0 and day 42; ACTH and PRL levels on day 42. Regression analysis of the 58 patients showed that on day 0, PRL levels were dependent on the ACTH plasma levels on day 0, the time of the collection of the blood sample and the age. On day 42, the PRL levels were dependent on the ACTH levels and the time of the blood collection but not on the age. Patients with lower baseline ACTH levels on day 0 displayed a better clinical outcome when taking the combination of milnacipran and pindolol as shown in the differences in MADRS on day 42. The same group of patients showed lower PRL levels on day 42. CONCLUSIONS: ACTH plasma levels at baseline or screening may help to predict the response to antidepressant treatment. PMID- 14533142 TI - Milnacipran for the treatment of chronic pain. PMID- 14533143 TI - Trichotillomania: response to lithium in a person with comorbid bipolar disorder. PMID- 14533147 TI - Metal-ion-binding peptides: from catalysis to protein tagging. PMID- 14533148 TI - Magnetic control of electrocatalytic and bioelectrocatalytic processes. AB - Bioelectronics is a rapidly progressing interdisciplinary research field that has important implications for the development of biosensors, biofuel cells, biomaterial-based computers, and bioelectronic devices. Magneto-controlled molecular electronics and bioelectronics are new topics that examine the effect of an external magnetic field on electrocatalytic and bioelectrocatalytic processes of functionalized magnetic particles associated with electrodes. In this article we describe the progress in the developments of magneto-switchable electrocatalytic and bioelectrocatalytic transformations, and the effects of the rotation of the magnetic particles on the electrocatalytic and bioelectrocatalytic processes are discussed. Finally, the implications of the results on the development of biosensors, amplified immunosensors, and DNA sensors are described. PMID- 14533149 TI - Molybdenum and tungsten imido alkylidene complexes as efficient olefin-metathesis catalysts. AB - Catalytic olefin metathesis has quickly emerged as one of the most often-used transformations in modern chemical synthesis. One class of catalysts that has led the way to this significant development are the high-oxidation-state alkylidene complexes of molybdenum. In this review key observations that resulted in the discovery and development of molybdenum- and tungsten-based metathesis catalysts are outlined. An account of the utility of molybdenum catalysts in the synthesis of biologically significant molecules is provided as well. Another focus of the review is the use of chiral molybdenum complexes for enantioselective synthesis. These highly efficient catalysts provide unique access to materials of exceptional enantiomeric purity and often without generating solvent waste. PMID- 14533150 TI - A covalent tetrathiafulvalene-tetracyanoquinodimethane diad: extremely low HOMO LUMO gap, thermoexcited electron transfer, and high-quality langmuir-blodgett films. PMID- 14533152 TI - First structurally authenticated zinc alkylperoxide: a model system for the epoxidation of enones. PMID- 14533151 TI - Design of environment-sensitive supramolecular assemblies for intracellular drug delivery: polymeric micelles that are responsive to intracellular pH change. PMID- 14533153 TI - Terpenoid-derived sulfides as ultimate organic sulfur compounds in extensively desulfurized fuels. PMID- 14533154 TI - One-pot synthesis of hierarchically ordered porous-silica materials with three orders of length scale. PMID- 14533155 TI - Synthesis and characterization of a cobalt(II) single-molecule magnet. PMID- 14533156 TI - A programmable one-pot oligosaccharide synthesis for diversifying the sugar domains of natural products: a case study of vancomycin. PMID- 14533157 TI - Rapid diversity-oriented synthesis in microtiter plates for in situ screening: discovery of potent and selective alpha-fucosidase inhibitors. PMID- 14533158 TI - Dormant states of rhodium hydroformylation catalysts: carboalkoxyrhodium complex formed from enones in the alkene feed. PMID- 14533159 TI - Measuring reaction kinetics by using multiple microcoil NMR spectroscopy. PMID- 14533160 TI - Rhodium-catalyzed Mizoroki-Heck-type arylation of alkenes with aroyl chlorides under phosphane- and base-free conditions. PMID- 14533161 TI - A practical solid-phase synthesis of glycosylphosphatidylinositol precursors. PMID- 14533162 TI - Transition-metal variation as a probe of the origins of hypoelectronic metallaboranes: eight- and ten-vertex open ruthenaboranes. PMID- 14533163 TI - Sequential Wittig olefination-catalytic asymmetric epoxidation with reuse of waste Ph3P(O): application of alpha,beta-unsaturated N-acyl pyrroles as ester surrogates. PMID- 14533164 TI - Total synthesis of (+)-wailupemycin B. PMID- 14533165 TI - Poly(methylene amine): a polymer with the maximum possible number of amino groups on a polymer backbone. PMID- 14533166 TI - The trifluoromethyltrioxy radical, CF3OOO. PMID- 14533167 TI - From particle-assisted wetting to thin free-standing porous membranes. PMID- 14533168 TI - Mesoscopic rings by controlled wetting of particle imprinted templates. PMID- 14533169 TI - Investigation of reactive intermediates of chemical reactions in solution by electrospray ionization mass spectrometry: radical cation chain reactions. PMID- 14533171 TI - Access of autonomic nerves through the optic canal, and their orbital distribution in man. AB - The notion that autonomic nerves from the internal carotid plexus are transmitted to the orbit with the ophthalmic artery through the optic canal has been variously assumed, disregarded, or denied, but never demonstrated. The objective of this study was to examine the contents of the canal, identify any autonomic nerves, and follow their passage within the orbit. The soft tissues of the optic canal, and the apical tissues of the orbit were removed and examined histologically using 10 cadaver preparations. Additionally, tissues from an orbital exenteration and 10 ocular enucleation or donor specimens were prepared. Some of the latter material was examined with an electron microscope. Numerous autonomic nerves (four to 25, ranging in diameter from 23 to 130 microm) entered the orbit from the internal carotid plexus in the periosteum of the optic canal, the optic nerve dura mater, or the adventitia of the ophthalmic artery. In the orbit they advanced in the loose connective tissue covering the optic nerve dura and joined ciliary nerves close to the eye or entered the eye directly. None were observed to penetrate the dura, apart from a nerve accompanying the central retinal artery. Others were distributed with the ophthalmic artery and its branches. It is concluded that the optic canal is a regular, and often major, route for autonomic nerve distribution to the eye and orbit. PMID- 14533172 TI - Morphological changes induced in the pig kidney by extracorporeal shock wave lithotripsy: nephron injury. AB - While shock wave lithotripsy (SWL) is known to cause significant damage to the kidney, little is known about the initial injury to cells along the nephron. In this study, one kidney in each of six juvenile pigs (6-7 weeks old) was treated with 1,000 shock waves (at 24 kV) directed at a lower pole calyx with an unmodified HM-3 lithotripter. Three pigs were utilized as sham-controls. Kidneys were fixed by vascular perfusion immediately after SWL or sham-SWL. Three of the treated kidneys were used to quantitate lesion size. Cortical and medullary samples for light (LM) and transmission electron microscopy (TEM) were taken from the focal zone for the shock waves (F2), the contralateral kidney, and the kidneys of sham-SWL pigs. Because preservation of the tissue occurred within minutes of SWL, the initial injury caused by the shock waves could be separated from secondary changes. No tissue damage was observed in contralateral sham-SWL kidneys, but treated kidneys showed signs of injury, with a lesion of 0.2% +/- 0.1% of renal volume. Intraparenchymal hemorrhage and injury to tubules was found at F2 in both the cortex and medulla of SWL-treated kidneys. Tubular injury was always associated with intraparenchymal bleeding, and the range of tissue injury included total destruction of tubules, focal cellular fragmentation, necrosis, cell vacuolization, and membrane blebbing. The initial injury caused by SWL was cellular fragmentation and necrosis. Cellular vacuolization, membrane blebbing, and disorganization of apical brush borders appear to be secondary changes related to hypoxia. PMID- 14533173 TI - Cartilage-free areas in the elbow joint of young golden retrievers. AB - The present study describes cartilage-free areas on the ulnar trochlear notch and the humeral condyle of eight very young golden retrievers with otherwise healthy elbow joints. Remarkably, the youngest dog with full-thickness cartilage-free areas was only 8 weeks old. The younger dogs showed no macroscopic abnormalities on the locations that were affected in the older dogs. Two kinds of cartilage modifications were found. Cartilage-free areas at the edges of the articular cartilage layer were present on the humeral capitulum and on two locations of the ulna, (the medial and lateral at the base of the anconeal process, and the trochlear notch near the lateral coronoid process, which was fractured in two cases). Histological examination showed that these cartilage-free areas were filled with dense supportive tissue. Synovial cells covered this tissue as well as the surrounding hyaline cartilage. The synovial membrane covering the areas was macroscopically enlarged, but histological examination revealed no signs of inflammation. The second type of modification consisted of discoloration of the articular surface at the humeral trochlea. Histological examination revealed that in this area the articular surface was composed of fibrocartilage instead of hyaline cartilage. Apparently, there are locations within the elbow joint in which articular cartilage is not necessary for normal joint functioning. The presence of fibrocartilage on the articular surface of the humeral condyle is a surprising finding, for which no explanation has yet been found. PMID- 14533174 TI - Normal ontogeny of perineal muscles and testosterone levels in Mongolian gerbils; response to testosterone in developing females. AB - The spinal nucleus of the bulbocavernosus (SNB) of Mongolian gerbils (Meriones unguiculatus) becomes sexually dimorphic during postnatal life, rather than prenatally as in rats. We therefore examined the early postnatal ontogeny of Mongolian gerbils, focusing on growth, serum testosterone (T) levels, and the sexually dimorphic perineal musculature innervated by the SNB. Serum T levels were higher in males than in females from birth through adulthood, with several early postnatal peaks and a large increase in T occurring during puberty in males. The SNB target muscles-the bulbocavernosus (BC) and levator ani (LA)-were present in both sexes on postnatal day 1 (PND1). Cross-sectional areas of BC fibers in males increased with age, and concurrently the myofibers of the BC became more fully developed and organized. In PND10 female pups, the BC muscle was virtually absent, while the LA muscle remained (although it was reduced in size). Postnatal treatment of female gerbils with androgen caused the BC muscle to remain and the LA muscle to become larger by PND10. Sexual dimorphism of the SNB develops differently in gerbils compared to other species, although its target muscles appear to respond to androgen in a manner similar to that in rats. PMID- 14533175 TI - Spatiotemporally separated cardiac neural crest subpopulations that target the outflow tract septum and pharyngeal arch arteries. AB - We used lacZ-retrovirus labeling combined with neural crest ablation in chick embryos to determine whether the cardiac neural crest cells constitute one group of multipotent cells, or they emigrate from the neural tube in time-dependent groups with different fates in the developing cardiovascular system. We demonstrated that early-migrating cardiac neural crest cells (HH9-10) massively target the aorticopulmonary septum and pharyngeal arch arteries, while the late migrating cardiac neural crest cells (HH12) are restricted to the proximal part of the pharyngeal arch arteries. These results suggest a prominent role for early migrating cells in outflow tract septation, and a function for late-migrating cells in pharyngeal arch artery remodeling. We demonstrated in cultures of neural tube explants an intrinsic difference between the early and late populations. However, by performing heterochronic transplantations we showed that the late migrating cardiac neural crest cells were not developmentally restricted, and could contribute to the condensed mesenchyme of the aorticopulmonary septum when transplanted to a younger environment. Our findings on the exact timing and migratory behavior of cardiac neural crest cells will help narrow the range of factors and genes that are involved in neural crest-related congenital heart diseases. PMID- 14533176 TI - Hepatic microvascular development in relation to the morphogenesis of hepatocellular plates in neonatal rats. AB - The development of hepatic microvascular heterogeneity after birth, and its temporal relationship to the development of parenchymal cell plates have received little attention. As a result, the morphogenesis of some of the parameters contributing to this heterogeneity in suckling and weaned rats was studied as a function of time between postpartum days 4 and 30 using in vivo light microscopic, electron microscopic, and immunocytochemical methods. During the early suckling period, the sinusoid network is highly anastomotic, with little evidence of zonation, and the parenchymal cell plates contain multiple cells and are irregularly arranged throughout the lobule. Sinusoidal endothelial fenestration is sparse at 4 days, but phagocytic Kupffer cell (KC) function already exists and exhibits zonal heterogeneity, with more cells located in the periportal zone. With increasing age, endothelial fenestrae increase and organize as sieve plates. Widened centrilobular radial sinusoids form through a loss ("drop-out") of intersinusoidal sinusoids (ISS). Concomitantly, the associated cell plates straighten and become one cell thick. Hepatocyte DNA synthesis and mitosis are higher in the periportal zone, which retains thickened cell plates and anastomotic sinusoids. The centrilobular sinusoids may widen to accommodate the increased volume of blood that results from the loss of ISS as well as the increased numbers of periportal sinusoids containing flow that feed these vessels. KC phagocytic activity increases during the suckling period concomitant with an increase of gut-derived endotoxin in the portal blood, which suggests that the KCs may be releasing mediators that affect sinusoid diameter, blood flow, endothelial fenestration, and perhaps parenchymal growth either directly or through the stimulation of growth factors. PMID- 14533177 TI - Innervation of the levator ani and coccygeus muscles of the female rat. AB - In humans, the pelvic floor skeletal muscles support the viscera. Damage to innervation of these muscles during parturition may contribute to pelvic organ prolapse and urinary incontinence. Unfortunately, animal models that are suitable for studying parturition-induced pelvic floor neuropathy and its treatment are rare. The present study describes the intrapelvic skeletal muscles (i.e., the iliocaudalis, pubocaudalis, and coccygeus) and their innervation in the rat to assess its usefulness as a model for studies of pelvic floor nerve damage and repair. Dissection of rat intrapelvic skeletal muscles demonstrated a general similarity with human pelvic floor muscles. Innervation of the iliocaudalis and pubocaudalis muscles (which together constitute the levator ani muscles) was provided by a nerve (the "levator ani nerve") that entered the pelvic cavity alongside the pelvic nerve, and then branched and penetrated the ventromedial (i.e., intrapelvic) surface of these muscles. Innervation of the rat coccygeus muscle (the "coccygeal nerve") was derived from two adjacent branches of the L6 S1 trunk that penetrated the muscle on its rostral edge. Acetylcholinesterase staining revealed a single motor endplate zone in each muscle, closely adjacent to the point of nerve penetration. Transection of the levator ani or coccygeal nerves (with a 2-week survival time) reduced muscle mass and myocyte diameter in the iliocaudalis and pubocaudalis or coccygeus muscles, respectively. The pudendal nerve did not innervate the intrapelvic skeletal muscles. We conclude that the intrapelvic skeletal muscles in the rat are similar to those described in our previous studies of humans and that they have a distinct innervation with no contribution from the pudendal nerve. PMID- 14533178 TI - Partial cranium of Cercopithecoides kimeui Leakey, 1982 from Rawi Gully, southwestern Kenya. AB - The Rawi Gully, located on the Homa Peninsula in southwestern Kenya, has produced several fossil elements of a large cercopithecid from sediments approximately 2.5 million years old (Ma). Nearly all of these elements appear to represent a single adult male individual of the colobine species Cercopithecoides kimeui Leakey, 1982. Part of the face, mandible, dentition, and several small postcranial fragments were collected by the Homa Peninsula Paleoanthropological Project (HPPP) in 1994 and 1995. This individual also appears to be represented by material collected in two previous expeditions to the site, one led by David Pilbeam in the 1970s and an earlier expedition led by L.S.B. Leakey in 1933. This specimen may extend the first appearance of C. kimeui by approximately 500 Kyr, and provides the first evidence for much of the male facial morphology in this species. Furthermore, Rawi may represent a more wooded habitat than the other occurrences of C. kimeui at Olduvai Gorge, Tanzania, and Koobi Fora, Kenya, indicating that C. kimeui may have been relatively flexible in its habitat preferences. PMID- 14533179 TI - Mobility in Upper Paleolithic and Mesolithic Europe: evidence from the lower limb. AB - A growing body of archeological evidence suggests that the dramatic climatic events of the Last Glacial Maximum in Europe triggered important changes in foraging behavior, involving a significant decrease in mobility. In general, changes in mobility alter patterns of bending of the midshaft femur and tibia, resulting in changes in diaphyseal robusticity and shape. This relationship between levels of mobility and lower limb diaphyseal structure was used to test the hypothesized decrease in mobility. Cross-sectional geometric data were obtained for 81 Upper Paleolithic and Mesolithic European femora and tibiae. The sample was divided into three time periods: Early Upper Paleolithic (EUP), Late Upper Paleolithic (LUP), and Mesolithic (Meso). In addition, because decreased mobility often results in changes in sex roles, males and females were analyzed separately. All indicators of bending strength decrease steadily through time, although few of the changes reach statistical significance. There is, however, a highly significant change in midshaft femur shape, with LUP and Meso groups more circular in cross-section than the EUP sample, supporting archeologically based predictions of decreased mobility. Sexual dimorphism levels in diaphyseal strength remain low throughout the three time periods, suggesting a departure in Upper Paleolithic and Mesolithic foragers away from the pattern of division of labor by sex observed in modern hunter-gatherers. Results confirm that the onset of the Last Glacial Maximum represents a crucial stage in Late Pleistocene human evolution, and signals the appearance of some of the behavioral adaptations that are usually associated with the Neolithic, such as sedentism. PMID- 14533180 TI - Matrix decomposition model for investigating prehistoric intracemetery biological variation. AB - A matrix decomposition model for analyzing prehistoric intracemetery biological variability is presented. The model, based on the correlation between interindividual biological and burial distance matrices, provides an estimate of the number of distinct burial populations interred within a single cemetery, which effectively ameliorates identifiability problems typical of general variability analyses (Raemsch [1995] Ph.D. dissertation, SUNY at Albany). Unlike other methods for analyzing intrasite variability analysis (e.g., kinship analysis and microchronology), this method is not reliant on a priori subgroup definition. Assuming a kin-structured burial pattern, a residual matrix is defined based on the raw data matrices; the proportion of negative or zero residuals to the total number of residuals provides an estimate of the number of hidden subgroupings (lineages) within the cemetery. The comparative utility of the model is demonstrated on a series of protohistoric and historic period skeletal samples from the Georgia coast, where it demonstrated that the increase in phenotypic variability during the early mission period is the result of population aggregation at the missions and not necessarily genetic admixture between local and nonlocal groups. PMID- 14533181 TI - Probable destructive meningioma in an archaeological adult male skull from Alaska. AB - Several intracranial pathological conditions can affect the bones of the skull. The most common cause of these conditions is tumor, but infection and other diseases are also known to affect the bones of the skull. Distinguishing between the various causes of intracranial skeletal pathology in archaeological human remains is usually a challenging exercise, and a specific diagnosis will often be impossible. Meningiomas are tumors that arise in arachnoid tissues embedded in the outer layer of the dura. Because of this association, they occur almost exclusively in the skull and vertebral column. Usually meningiomas are slow growing tumors that do not metastasize to other organs and tissues of the body. However, rare cases can be malignant and, even when meningiomas are benign, their presence and growth can adversely affect the nervous and vascular supply to other tissues in the skull and vertebral column. Their effect on adjacent bone tissue varies from stimulating bone-forming lesions to causing highly destructive lesions. A few examples of meningioma have been described in the paleopathological literature. Most of these cases are bone-stimulating meningiomas. The case presented here is a probable example of a highly destructive meningioma of the skull base, with unilateral extension into the left side of the cranium. This case is compared with a modern clinical case of destructive intracranial meningioma that was documented both radiographically and pathologically. Destructive meningiomas can be confused with other pathological conditions, including benign and malignant tumors. Criteria for differentiating the diagnostic options are reviewed. PMID- 14533182 TI - Predicting nose projection and pronasale position in facial approximation: a test of published methods and proposal of new guidelines. AB - Many prediction guidelines exist in facial approximation for determining the soft tissue features of the face, and the reliability of each is generally unknown. This study examines four published and commonly used soft-tissue prediction guidelines for estimating nose projection, two of which also estimate the position of the pronasale. The methods tested are those described by: 1) Gerasimov ([1971] The Face Finder; London: Hutchinson & Co.), using the distal third of the nasal bones and the nasal spine; 2) Krogman ([1962] The Human Skeleton in Forensic Medicine; Springfield: Charles C. Thomas), using the average soft-tissue depth at midphiltrum, plus three times the length of the nasal spine (and a variation of this technique: plus three times the distance of the tip of the nasal spine from the nasal aperture); 3) Prokopec and Ubelaker ([2002] Forensic Sci Commun 4:1-4), using the reflected profile line of the nasal aperture; and 4) George ([1987] J Forensic Sci 32:1305-1330), using a variation of the Goode method. Four identical hard-tissue tracings were made of 59 adult lateral head cephlograms (29 males, mean age 24, SD 10 years; 30 females, mean age 23, SD 5 years) on separate sheets of tracing paper. One soft-tissue tracing was also made for each radiograph. All tracings were marked with three identical reference points. Soft-tissue tracings were isolated from one of us (C.N.S.), who attempted under blind conditions to predict pronasale position and nose projection on the hard-tissue tracings, using the soft-tissue prediction guides above. Actual soft-tissue tracings were then compared to each of the predicted tracings, and differences in projection/pronasale position were measured. Results indicate that for nose projection, methods 3 and 4 performed well, while methods 1 and 2 performed poorly. Features which are most related to nose projection/pronasale are described in this paper, as are regression equations generated from these variables that predict pronasale/nose projection better than the traditional methods mentioned above. The results of this study are significant because they: 1) indicate that the popular facial approximation methods used to build the nose are inaccurate and produce incorrect nose anatomy; and 2) indicate that the new pronasale prediction methods developed here appear to have less error than traditional methods. PMID- 14533183 TI - Pair-specific usage of sleeping sites and their implications for social organization in a nocturnal Malagasy primate, the Milne Edwards' sportive lemur (Lepilemur edwardsi). AB - Safe sleeping sites may be a limited resource crucial for survival. In order to investigate their potential significance for social organization in nocturnal primates, we analyzed the spatial distribution of daily sleeping sites, their characteristics, their usage, and sleeping group compositions in the nocturnal Milne Edwards' sportive lemur during a 6-month field study in the dry deciduous forest of northwestern Madagascar. Sexes did not differ either in body size or in body mass. Sleeping sites were used almost exclusively by adult male-female pairs. Individuals showed a high sleeping-site fidelity limited to 2-3 different sleeping sites in close vicinity during the whole study period. Most females showed a higher fidelity to one distinct sleeping site than their male partners. Sleeping groups consisted of one adult male and one adult female and remained stable in composition over the whole study period. Exclusive pair-specific usage of sleeping sites suggests sleeping site related territoriality of male-female pairs, perhaps influenced by inter- and intrasexual resource competition. Results give first insights into the distribution patterns and social organization of this species. They imply dispersed monogamy for the Milne Edwards' sportive lemur, with sleeping sites as a potentially restricted and defendable resource. PMID- 14533184 TI - Reconstruction of human evolutionary tree using polymorphic autosomal microsatellites. AB - Allelic frequencies of 182 tri- and tetra-autosomal microsatellites were used to examine phylogenetic relationships among 19 extant human populations. In particular, because the languages of the Basques and Hunza Burusho have been suggested to have an ancient relationship, this study sought to explore the genetic relationship between these two major language isolate populations and to compare them with other human populations. The work presented here shows that the microsatellite allelic diversity and the number of unique alleles were highest in sub-Saharan Africans. Neighbor-joining trees based on genetic distances and principal component analyses separated populations from different continents, and are consistent with an African origin for modern humans. For the first time, with biparentally transmitted markers, the microsatellite tree also shows that the San are the first branch of the human tree before the branch leading to all other Africans. In contrast to an earlier study, these results provided no evidence of a genetic relationship among the two language isolate groups. Genetic relationships, as ascertained by these microsatellites, are dictated primarily by geographic proximity rather than by remote linguistic origin, Mantel test, R(0) = 0.484, g = 3.802 (critical g value = 1.645; P = 0.05). PMID- 14533185 TI - Longitudinal analysis of deciduous tooth emergence: III. Sexual dimorphism in Bangladeshi, Guatemalan, Japanese, and Javanese children. AB - Previous studies, mostly in European populations, found sex differences in the pattern of deciduous tooth emergence. Most studies find that the anterior dentition in males is precocial relative to the female dentition, and the pattern reverses so that females lead males in the emergence of the posterior deciduous dentition. Less is known about sex differences in the dental development and emergence of non-European populations. Here we examine the pattern of sex differences in deciduous tooth emergence in Japanese, Javanese, Guatemalan, and Bangladeshi children. The data come from four longitudinal or mixed longitudinal studies using similar study protocols. Survival analysis was used to estimate parameters of a log-normal distribution of emergence for each of the 10 teeth of the left dentition, and sexual dimorphism was assessed by sex-specific differences in mean emergence times and by Bennett's index. The results support the pattern of developmental cross-over observed in other populations. We conclude that there is little evidence to support the hypothesis of Tanguay et al. ([1984] J. Dent. Res. 63:65-68) that ethnic factors mediate sex differences in the emergence of deciduous teeth. PMID- 14533186 TI - Marriage, parenting, and testosterone variation among Kenyan Swahili men. AB - Male variation in testosterone (T) levels may, in part, reflect a differential behavioral allocation to mating and parenting effort. This research tests whether demographic indicators of pair bonding and parenting were associated with salivary T levels among Kenyan Swahili men. Men in the sample were either unmarried (N = 17), monogamously married (N = 57), or polygynously married (N = 14), and between ages 29-52. In contrast with earlier findings among North American men, monogamously married men did not have lower T levels than unmarried men. However, among all married men, men with younger genetic children tended to have marginally lower T levels. Polygynously married men, all of whom had two wives, had higher T levels than all other Swahili men. Possible explanations of higher T levels among polygynously married men are explored. PMID- 14533187 TI - Remembering Malthus II: establishing sustainable population optimums. PMID- 14533188 TI - Remembering Malthus III: implementing a global population reduction. PMID- 14533191 TI - Rational development of a HIV-1 gene therapy vector. AB - BACKGROUND: HIV-1 provides an attractive option as the basis for gene transfer vectors due to its ability to stably transduce non-cycling cell populations. In order to fully utilise the promise of HIV-1 as a vector it is important that the effects of viral cis sequence elements on vector function are carefully delineated. METHODS: In this study we have systematically evaluated the effect of various cis elements from the HIV-1 YU-2 genome that have been implicated as either affecting vector performance, or HIV-1 replication, on the efficiency of vector production (titre and infectivity). As a measure of the relative safety of vectors their propensity to inadvertently transfer the gagpol gene to transduced cells was assessed. RESULTS: Sequences that were found to increase vector titre were from the 5' end of the gag gene, from the 5' and 3' ends of the env gene, from immediately upstream of the polypurine tract, and the central polypurine tract. The substitution of the HIV-1 RRE with heterologous RNA transport elements, or the deletion of the RRE, resulted in greatly reduced vector titres. RNA analysis suggested that the role of the Rev/RRE system extends beyond simply acting as an RNA nuclear export signal. The relative safety of different vector designs was compared and an optimal construct selected. CONCLUSIONS: Based on our results we have constructed a vector that is both more efficient, and has better safety characteristics, than the widely used pHR' HIV-1 vector construct. PMID- 14533192 TI - Modulation of renal glomerular disease using remote delivery of adenoviral encoded solubletype II TGF-beta receptor fusion molecule. AB - BACKGROUND: Systemic adenoviral (Ad) gene therapy for renal disorders is largely hampered by the unique architecture of the kidney. Consequently, currently available Ad vectors are of only limited therapeutic utility in the context of glomerular and fibroproliferative renal diseases. METHODS: The Ad vectors studied in the context of blocking renal fibrosis were AdTbeta-ExR and AdCATbeta-TR. AdTbeta-ExR encodes a chimeric soluble molecule comprising the entire ectodomain of the human type II TGF-beta receptor, genetically fused to the Fc fragment of the human IgG1 (sTbetaRII), while AdCATbeta-TR encodes only the dominant-negative truncated ectodomain of the human type II TGF-beta receptor. The biologic activity of the type II TGF-beta receptor was evaluated in vitro by its ability to inhibit cellular proliferation and in vivo in a unilateral ureter obstruction fibrosis model. Renal targeting with sTbetaRII was evaluated immunohistochemically after intramuscular (IM) delivery of AdTbeta-ExR. The renal antifibrotic effect of the Ad vectors was evaluated in a lupus murine model with both light and electron microscopy and urinalysis. RESULTS: sTbetaRII was detected in the glomeruli after remote IM injection of AdTbeta-ExR, but not the control AdCATbeta-TR, indicating renal deposition of the heterologous soluble fusion protein after its expression in the muscle and secretion into the circulation. AdTbeta-ExR, but not AdCATbeta-TR, could transiently inhibit mesangial expansion, glomerular hypercellularity, proteinuria and cortical interstitial fibrosis in a murine lupus model. However, the autoimmune renal disease eventually surpassed the antifibrotic effect. CONCLUSIONS: These results indicate the superiority of a soluble type II TGF-beta receptor over a dominant negative, non-soluble type II TGF-beta receptor in the context of blocking renal fibrosis in murine models. PMID- 14533193 TI - Chemotherapy increases transgene expression in leukemic cells. AB - BACKGROUND: Patients with acute myeloid leukemia (AML) often obtain complete remission with chemotherapy but the majority of patients relapse. Combining chemotherapy and gene therapy appears to be a promising approach; however, the effects of chemotherapy on transgene expression in leukemic cells have not yet been investigated. METHODS: DA1-3b leukemic cells were transfected with pCDNA3 plasmids carrying GM-CSF or LacZ cDNA. The leukemic K562 cell line and primary cultured AML cells were transduced with an Ad5.CMV-LacZ adenoviral vector. Cells were then incubated with various concentrations of daunorubicin (DNR) and cytosine arabinoside (Ara-C), and expression of the transgene was measured. Murine DA1-3b-pCDNA3/LacZ leukemic cells were also injected into syngeneic C3H/Hej mice. RESULTS: In the cells carrying pCDNA3, DNR and Ara-C dramatically increased expression of the LacZ and GM-CSF transgenes. Over-expression depended on drug dose and was due to increased transcription. Enhancement was also observed in K562 cells and in some primary cultured AML samples transduced with the Ad5.CMV-LacZ adenovirus. Addition of N-acetyl-L-cysteine inhibited the over expression, suggesting that reactive oxygen species were involved in activating the CMV promoter. In the A549 lung carcinoma cell line transduced with Ad5.CMV LacZ, Ara-C had only a minor effect, and DNR had a detrimental effect, suggesting that expression depends on cell type. In vivo experiments in which mice received DA1-3b-pCDNA3/LacZ leukemic cells, and were then treated with Ara-C, also showed increased transgene expression in these leukemic cells. CONCLUSIONS: In leukemic cells, chemotherapeutic agents can induce over-expression of transgenes. This suggests a promising combined strategy for the treatment of acute leukemia. PMID- 14533194 TI - Combination with CD/5-FC gene therapy enhances killing of human bladder-cancer cells by radiation. AB - BACKGROUND: Resistance to radiation and chemotherapy is a significant obstacle to the treatment of advanced bladder cancer. Gene therapy combined with radiation represents a new approach to cancer treatment. In the present study, we investigated whether adenovirally directed, cytosine deaminase (CD)/5 fluorocytosine (5-FC) gene therapy could induce cell toxicity and radiosensitization through the intracellular production of 5-fluorouracil (5-FU) in bladder-cancer cells. METHODS: Three human bladder-cancer cell lines, KK47 (wild-type p53+), T24 (p53 mutated) and 5637 (p53 mutated), were investigated. A recombinant adenovirus vector containing the CD gene (Ad-RSV-CD) was used. Cells were infected with Ad-RSV-CD and treated with 5-FC. Forty-eight hours after infection, the cells were irradiated and cytotoxicity assays performed to determine the extent of increase in in vitro cytotoxicity. A KK47 subcutaneous tumor-xenografts model was used in an animal study to examine the tumor growth inhibitory effect of this combination therapy. Ad-RSV-CD was directly injected into the tumor and daily 5-FC was intraperitoneally injected. Forty-eight hours after injection of Ad-RSV-CD, the tumor was irradiated. The tumor volume was measured every day. RESULTS: In all three cell lines, the combination treatment enhanced the cell killing of human bladder-cancer cells in vitro. It also enhanced the tumor-growth inhibition in the KK47 tumor model. CONCLUSIONS: In the present study, we demonstrated that CD/5-FC gene therapy combined with radiation therapy enhances cell killing of human bladder-cancer cells in in vitro and in vivo animal models. PMID- 14533195 TI - Enhancing adenovirus-mediated gene transfer in vitro and in vivo by addition of protamine and hydrocortisone. AB - BACKGROUND: Inclusion of positively charged polymers such as protamine in adenovector formulations has been reported to improve the efficiency of adenovirus-mediated gene transfer in vitro and in vivo. On the other hand, corticosteroids are known to inhibit inflammation and thus might be useful in minimizing vector-related toxicity. In this study, we evaluated the combined effect of protamine sulfate and hydrocortisone on the efficiency of adenovirus mediated gene transfer in vitro and in vivo. METHODS: Protamine and hydrocortisone at different concentrations were added to adenovector formulations. In vitro transgene expression with or without inclusion of protamine and hydrocortisone was evaluated in the breast cancer cell lines MDA-MB 231 and MCF7 and the lung cancer cell lines A549 and H460. In vivo transgene expression in the mouse lung was determined after aerosolized vector delivery. RESULTS: The combination of 2 micro g/ml protamine and 125 ng/ml hydrocortisone significantly increased transgene expression in vitro in all the cell lines tested. Protamine is only effective when it is added to cells before or together with adenovectors, whereas hydrocortisone is effective when it is added to cells before, together with, or after adenovectors. Inclusion of protamine and hydrocortisone also augmented apoptosis induction caused by adenovectors expressing proapoptotic genes in cancer cells. Moreover, protamine and hydrocortisone dramatically enhanced transgene expression in the mouse lung after aerosolized vector delivery. CONCLUSIONS: Inclusion of protamine and hydrocortisone in adenovector formulations can improve adenovector-mediated gene expression and may be useful for clinical applications of current adenovirus mediated gene therapy. PMID- 14533196 TI - Coxsackie-and-adenovirus receptor mRNA expression in human heart failure. AB - BACKGROUND: Adenoviral vectors are widely used as gene-transfer vehicles in experimental and clinical studies. Since virus incorporation and transfection efficacy depend to a large extent on the concentration of the coxsackie-and adenovirus (CAR) receptor on target cells the aim of this study was to quantify the CAR-receptor concentration in various human cardiomyopathies. METHODS: After RNA isolation from myocardial biopsies obtained during surgical procedures, cDNA was generated by reverse transcription. The relative RNA content was analyzed by quantitative PCR using glyceraldehydes-3-phosphate dehydrogenase (GAPDH) as a standard reference. The cardiomyopathies (CM) analyzed were categorized according to their etiology in dilated CM (DCM, n = 28), ischemic CM (ICM, n = 52), CM in mitral valve disease (MVCM, n = 32) and aortic valve disease (AVCM, n = 32). Data were related to non-cardiomyopathic tissue from donor hearts (non-CM, n = 64). RESULTS: Compared with non-CM hearts DCM showed a 34-fold (+/-5.4) increase in CAR mRNA concentration, in ICM CAR mRNA was elevated by a factor of 12 (+/-4.3), in MVCM by 27 (+/-7) and AVCM by factor 47 (+/-9.3) (ANOVA p < 0.001). Compared with the expression in rat hearts CAR levels were found to be similar to those in human ICM. CONCLUSIONS: These results show that cardiomyopathies associated with heart failure transcribe substantially higher levels - on average by a factor of 30 - of CAR-mRNA than non-failing control hearts. Myocardial gene transfer using adenoviral vectors should therefore be facilitated in human cardiomyopathies and may present a promising approach for therapeutic interventions. PMID- 14533198 TI - Enhanced antitumor activity of a combination of MBD2-antisense electrotransfer gene therapy and bleomycin electrochemotherapy. AB - BACKGROUND: MBD2 is a methylated DNA-binding protein that has been previously suggested to have transcriptional silencing as well as DNA demethylase activities. We have previously shown that electrotransfer of an MBD2-antisense encoding plasmid inhibits tumor growth in vivo. In this study we tested whether a combination of MBD2-antisense gene therapy and bleomycin chemotherapy has an augmented antitumor effect in comparison with either monotherapy. METHODS: Mice bearing human non-small-cell lung carcinoma line H1299 xenoplants were treated with electrotransfer of either bleomycin or MBD2-antisense expression plasmid or a combination of both therapies and tumor growth following treatment was monitored. RESULTS: A combination of electrotransfer of MBD2-antisense and bleomycin electrochemotherapy has an additive inhibitory effect on the rate of tumor growth and a synergistic effect on the number of tumor-free animals when compared with either monotherapy. CONCLUSIONS: Our results suggest that a combination of MBD2-antisense electrotransfer gene therapy and chemotherapy with bleomycin is a candidate new approach to anticancer therapy. PMID- 14533197 TI - Functional delivery of large genomic DNA to human cells with a peptide-lipid vector. AB - BACKGROUND: Nonviral gene transfer vectors have the potential to deliver much larger DNA constructs than current viral vectors but suffer from a low transfection efficiency. The LID vector, composed of Lipofectin (L), an integrin targeting peptide (I) and DNA (D), is a highly efficient synthetic vector, both in vitro and in vivo, which may allow the transfer of genomic loci for gene therapy. METHODS: Transfection efficiencies were quantitated using the green fluorescent protein (GFP) reporter. Expression of a large genomic locus (NBS1 [Nijmegen breakage syndrome], encoding nibrin) was assessed by immunofluorescence. RESULTS: We report a systematic study of the parameters influencing delivery of BAC-based plasmids ranging in size from 12 to 242 kb using the LID vector. We showed 60% of cells were transfected with the smaller plasmids while plasmids up to 242 kb were consistently delivered to over 10% of cells. The number of transfected cells was related to number of plasmids in the transfection complex independent of plasmid size. Atomic force microscopy showed that LID particle size increased with plasmid size consistent with one plasmid molecule per particle. When LID vectors were used to deliver the NBS1 gene as a 143 kb construct to primary NBS cells, at least 57% of cells expressing GFP also expressed functional nibrin. CONCLUSIONS: We show that LID vectors represent a promising tool for the transfer of complete genomic loci. PMID- 14533200 TI - DNA transfection of mononuclear cells in muscle tissue. AB - BACKGROUND: Genes encoding non-self proteins may be injected into skeletal muscles in vivo to obtain induction of cellular and humoral immune responses against the encoded antigens (DNA vaccination). Bone marrow derived professional antigen-presenting cells (APCs) play a key role in the induction of immunity by DNA vaccination. In the present work we have investigated whether the APCs are transfected by DNA injection into muscle. METHODS: DNA encoding green fluorescent protein (GFP) was injected into rat and mouse limb muscle and followed by electroporation. Whole mount muscle tissue with GFP-positive mononuclear cells (MNCs) were treated with immunocytochemical markers specific for leukocytes, and studied with fluorescent microscopy. To detect transfected cells migrating to peripheral lymphoid tissue RT-PCR was applied on RNA isolated from the draining popliteal lymph node and spleen. Lymphoid tissue was also analyzed with real-time PCR for distribution of the injected plasmid. RESULTS: MNCs were transfected after intramuscular DNA injection, and, following DNA injection with electroporation, the number of GFP-positive MNCs increased 6-fold in rats and 14 fold in mice. None of the GFP-positive MNCs were stained with leukocyte-specific antibodies. Even though GFP encoding DNA was detected in the popliteal lymph node, no RNA encoding GFP was found in the lymph node or spleen. However, MHC II positive cells in the muscle tissue appeared preferentially around the transfected MNCs. CONCLUSIONS: Many MNCs in the muscle are transfected after intramuscular DNA injection. Electroporation significantly increases the number of transfected MNCs. None of the observed transfected MNCs however were leukocytes. MHC II-positive cells accumulated around transfected MNCs; this suggests that transfer of antigen from transfected MNCs to APCs may contribute to the immune response. PMID- 14533199 TI - Myocardial injection of CA promoter-based plasmid mediates efficient transgene expression in rat heart. AB - BACKGROUND: Although naked plasmid injection is the safest and most convenient method for gene delivery, a major limitation of this approach is currently poor transgene expression. The CA promoter (chicken beta-actin promoter with cytomegalovirus, CMV, enhancer) is one of the strongest transcriptional control modules found; however, it is uncertain whether a CA promoter-based vector is efficient enough for naked gene therapy in a cardiovascular context. METHODS: The beta-galactosidase (LacZ) expression provided by CA promoter plasmid (pCAZ2) injection into the skeletal muscle or the heart of Lewis rats was compared with CMV promoter plasmid or adenoviral vector (AxCAZ3). The effect of Simian virus 40 of the replication origin (SV40ori) deletion from pCAZ2 on transgene expression was also evaluated. RESULTS: pCAZ2 showed the highest LacZ expression in both skeletal muscle and heart in comparison with the CMV promoter-based vector 5 days after naked plasmid injection. LacZ expression in the heart obtained using 20 micro g of pCAZ2 was almost equivalent to that shown with AxCAZ3 at 6.0 x 10(9) optical particle units. The time course of transgene expression driven by CMV and CA promoters in the heart were similar, with the CA promoter providing significantly higher gene expression than the CMV promoter across all time points examined. SV40ori deletion from pCAZ2 did not affect transgene expression in either skeletal muscle or heart. CONCLUSIONS: Transgene expression mediated by naked CA promoter-based plasmid injection was shown to be quite efficient in the heart. We propose that the CA promoter vector is suitable for myocardial gene therapy. PMID- 14533203 TI - How much does it hurt? The pervasive problem of pain. PMID- 14533204 TI - International Cancer Research Portfolio: new on-line tool will aid in cancer research collaboration and planning. PMID- 14533206 TI - Workplace issues of HIV-positive nurses. PMID- 14533208 TI - Nursing workforce supply and demand: implications for nursing education capacity and the health care system in Colorado. PMID- 14533209 TI - Know your staff numbers--and know you're right. PMID- 14533210 TI - Improving end-of-life care in the intensive care unit: are nurses involved? PMID- 14533211 TI - The closed tracheal suction catheter: 24 hour or 48 hour change? AB - Closed suction catheters (CSC) for removal of bronchial secretions in intubated patients have been used in intensive care units (ICU) for many years. Manufacturers still recommend daily changes of the catheter in order to reduce the incidence of ventilator associated pneumonia (VAP). There is, however, a lack of clinical evidence to support this recommendation. The objective of this study was therefore to compare the incidence of VAP in patients who receive either 24 hourly or 48 hourly changes of the CSC. Eligible patients were randomised to one of the two groups to receive either a 24 hourly change (n = 53) or a 48 hourly change (n = 48) of the CSC. Sputum specimens were sent second daily for quantitative culture. Chest x-rays (CXR) and white blood cell counts were attended daily. A VAP was diagnosed according to previously established criteria. A second set of modified criteria were also used to conduct a further analysis of the results. Of the 158 patients randomised, 101 completed the study. These patients had a mean age of 65 years and a mean APACHE II score of 28.2 in the first 24 hours of the study. The average duration in the trial was 10 days. The two groups were comparable in terms of demographic features. There were no reported cases of VAP in either group using the criteria originally selected in the study design. Using a modified criteria to diagnose VAP there were 10 (19%) patients with VAP in the 24 hour group and 13 (27%) in the 48 hour group. The incidence of VAP between the two groups was not statistically different (p = 0.35). To conclude, there was no difference in the incidence of VAP between the two groups studied. Based on previous studies conducted by Quirke and Kollef and the experience of our study we have changed our clinical practice to a 48 hour change of the CSC. We would, however, suggest further study or a meta-analysis of the available literature before a recommendation is made. PMID- 14533212 TI - Physical stresses related to the transport of the critically ill: optimal nursing management. AB - The critically ill patient's physical well being is constantly at risk. This fragile physical state means the patient is vulnerable to a host of hazards within the critical care setting, none of which are more relevant to patient outcome than the hazards awaiting them should the patient require transportation. The stresses exerted on the human body and the vulnerability of the critically ill to various means of movement, transport and subsequent environmental changes are explored through this review. The review includes a brief history of the evolution of patient transportation, which has provided a basis for the inception of formal strategies in patient management related to transportation. Within this framework current optimal nursing management minimising the detrimental effects of transport on the critically ill is reasoned to its scientific base. The basic laws of physics are an important consideration in all modes of patient movement or transport and play an integral role in the critical care nurses' practice. A clear understanding of pathophysiological and technological processes involved in caring for the critically ill and applying the principles of physics to effective contextual practice enhances the capability of the critical care nurse. PMID- 14533213 TI - Chest x-ray quiz. PMID- 14533214 TI - Oral care of the critically ill: a review of the literature and guidelines for practice. AB - Maintaining oral health in the critically ill patient is imperative in reducing the risk of nosocomial infections and improving patient comfort and discharge outcomes. Critically ill patients are at great risk for poor oral health as many are elderly, undernourished, dehydrated, immunosuppressed, have a smoking or alcohol history, are intubated or on high-flow oxygen, and are unable to mechanically remove dental plaque. Many modalities for delivering oral care have been reported in the literature. The use of the toothbrush in the mechanical removal of plaque, even in the intubated patient, has been proven to be superior to the swab. Brushing of the gums in edentulous patients is of benefit. Although electric toothbrushes are preferable, their cost, size and the potential for cross-infection limits their use. Chlorhexidine has long been the gold standard for mouthwashes and provides up to 24 hours of antimicrobial activity; therefore infrequent applications are adequate. Sodium bicarbonate and hydrogen peroxide are of limited use due to lack of convincing evidence regarding their safety and antimicrobial effects in the critically ill population. Saliva stimulants or substitutes including lemon and glycerine are also inappropriate for moistening the oral cavity in the critically ill patient. Regular oral assessment and individualized oral care, along with the use of a standardised protocol for oral care (incorporating proven modalities) is vital for optimal oral care in the critically ill patient. PMID- 14533215 TI - The ventilated patient undergoing hydrotherapy: a case study. AB - The ascending peripheral neuropathy and paralysis that result from Guillain-Barre Syndrome's (GBS) demyelination of peripheral nerves is a challenge to health professionals; the patient requires support during the acute disease process and during the remyelination recovery period, often lasting months to years. The staff of a major metropolitan teaching hospital's critical care unit (CCU) and physiotherapy departments developed a hydrotherapy treatment programme for a ventilated patient with GBS. Through careful planning and appropriate preparation, it was found that hydrotherapy could successfully and safely be incorporated into a patient's treatment regimen. The benefits included improved range of movement due to the supportive nature of water, anecdotal increased strength, size and movement of remyelinating muscles and a psychological improvement. Although this patient has not recovered from GBS to be independent, hydrotherapy was a valuable part of the treatment regimen and it could be suggested the increase muscle strength lead to improved respiratory function and enabled weaning from ventilation, reducing intensive care length of stay and cost. PMID- 14533216 TI - The long haul. PMID- 14533217 TI - Let us get on with it. PMID- 14533218 TI - In the blood. PMID- 14533219 TI - The CJD mystery. PMID- 14533220 TI - Too clever by half. PMID- 14533221 TI - Reading between the lines. PMID- 14533222 TI - 'My son is an i.v. drug user'. PMID- 14533223 TI - Modernization: the role of ward sisters and charge nurses. AB - BACKGROUND: The nursing profession is being directed by the government to modernize alongside the NHS as an organisation (DoH 2000). The role of ward sister/charge nurse has taken the brunt of increasing managerialization and the reduction in junior doctors' hours. This article examines how the role has developed and explores future opportunities. CONCLUSION: The ward sister/charge nurse role is pivotal to the modernization of the NHS. However, unless the role, responsibilities and expectations of these experienced nurses are clearly defined, the position may cease to be a promotional aspiration of junior nurses. The negative consequences of this for standards of patient care and recruitment and retention of staff are likely to be significant. PMID- 14533224 TI - Nurse education: why Socrates would disapprove. PMID- 14533225 TI - Scar management. AB - Scarring has major psychological and physical repercussions--for example, scarring on the face and visible regions of the body can be very distressing for the patient, whether it is simple acne scars or large, raised surgical or traumatic scars. This article discusses the process of scar formation, the differences between scars and proposes a number of ways in which the nurse can manage scars. PMID- 14533226 TI - Effective interventions for pressure ulcer prevention. AB - This article examines the causes of pressure ulcers and provides an overview of the best advice available in preventing them in the clinical setting. This should enable nurses to provide more effective interventions for preventing patients from developing pressure ulcers. PMID- 14533227 TI - More than cosmetic. PMID- 14533228 TI - Asking for help. PMID- 14533229 TI - Understanding the biological bases of mental illness. PMID- 14533230 TI - Becoming a mental health nurse: a qualitative study [Part-2]. AB - This is a two-part commentary reporting on a qualitative piece of research concerning the experiences of student nurses and the processes involved in becoming a mental health nurse (Morrissey 2003). It is really an attempt to describe, connect, discuss, and understand the process of becoming a mental health nurse in theory and practice using a qualitative approach (Holliday, 2002; Field & Morse, 1994; Hammersley, 1992; Burnard, 1991; Glaser & Strauss, 1967;). A questionnaire and semi-structured interviews were employed to examine the student nurses experiences prior to and one year into their first staff nurse post. The term psychiatric nurse and mental health nurse continue to be used interchangeably in practice and in written texts. However for simplicity the term mental health nurse is used for this text. PMID- 14533231 TI - Social constructionism, discourse analysis and mental health nursing: a natural synergy. AB - This paper has been developed to identify the natural synergy between social constructionism, discourse analysis and mental health research. It is based on research undertaken to explore mental health nurses' identity. The proposal is that nurses' identities are rhetorically constructed in the language they use to account for and justify their work in the practice context. PMID- 14533232 TI - Mentor relationship as a tool of professional development of student nurses in clinical practice. AB - This is a condensed version of a research project relating to the design and the development of a research instrument concerning 'Mentor Relationship as a Tool of Professional Development of Student Nurses in Clinical Practice'. This short research paper is taken from and reproduces the research work undertaken by Saarikoski (2002). The main themes refer to: (1) Evaluation scale to assess the quality of clinical learning environment and (2) Supervision of student nurses during their clinical placements. Parts one and two are taken from the main research study and include the following (i) developing and testing an evaluation tool (ii) describing how nursing students experience their clinical learning environment and (iii) the supervision given by qualified staff nurses working in a hospital setting. This abridged report discusses the methodology approaches undertaken by the author and includes: (a) comparative phased twin centred study (b) a pilot scheme and (c) a primary research instrument that was developed into an extensively validated assessment-measuring tool. This report strongly suggests that there is clear evidence in this research report that the supervisory relationship is the most important single element of pedagogical activities of staff nurses. The total satisfaction of students correlated most clearly with the method of supervision and that those satisfied students had a successful mentor relationship and frequently enough access to private supervision sessions with mentor. In the sample of this empirical study (n = 279 student nurses in Finland) individualized supervision system was most common on psychiatric wards. All nurse educators and clinical practitioners working across Europe and around the World in clinical learning environments will find this paper very useful in helping them to improve and quantify the supervisory process. This study starts bridging the gap between using and integrating both at a National and European level qualitative assessment systems that relate to the learning and supervisory process. The study encourages the need for professionals to test these new instruments in other nursing cultures and reflects upon the need for further research work in this area. PMID- 14533233 TI - Treatment for substance abuse in Australia: a comparison of public and private programs. AB - Alcohol and drug abuse is a major problem in Australia and caused over 4000 deaths in 1998 alone. The national policy for reducing the harmful consequences of drug and alcohol use focuses on decreasing the production and supply of drugs, reducing the demand for drugs including a focus on abstinence in treatment, and minimizing the harm to individuals and communities. This research examines substance abuse treatment in Australia, and identifies similarities and differences in funding, philosophy, purpose, and strategies between public and private treatment programs. Interviews of 21 treatment program directors in Sydney and Melbourne were done, using a valid and reliable semi-structured interview guide. Nine public, not-for-profit programs primarily focused on consultation to medical providers, detoxification, outpatient counseling, and harm minimization including methadone treatment and needle exchange. Rehabilitation, prevention, and/or research were objectives of a limited number. Twelve private, non-governmental programs primarily provided abstinence-based treatment in a variety of settings. Some for-profit programs, funded by insurance, provided three to four weeks inpatient care, usually with outpatient follow-up. Not-for-profit programs provided residential treatment in a therapeutic community over a period of 6 months to 2 years. Still others (both profit and not-for-profit) provided methadone treatment or outpatient treatment. Four were church related and six received funding through government contracts, social security illness benefits, and housing assistance. The general lack of structured rehabilitation programs seemed to be the greatest weakness of the Australian public programs. A low percentage of patients completing detoxification entered counseling, and studies were not available to show the outcomes of detoxification without follow-up. The availability of consultations to hospitals and health care practitioners seemed to be an excellent advantage, but the lack of structured SAT beyond detoxification may have minimized that advantage. Nurses were involved in a variety of roles in Australia, and six of the program directors or managers we interviewed were nurses. Nurses were usually in clinical roles or management roles, rather than counseling roles. PMID- 14533234 TI - Importance of holistic nutritional assessment in wound healing. PMID- 14533235 TI - An evaluation of the leg and foot ulcer treatment provided in Oslo. AB - OBJECTIVE: This study set out to gather more information about the treatment of leg and foot ulcers in the community in Oslo, Norway. METHOD: A structured questionnaire on leg and foot ulcer treatment was sent to 173 community nurses in Oslo, of whom 102 responded with data on 145 patients. RESULTS: Of the sample, 55.9% had leg ulcers, 32.2% had foot ulcers and 11.9% both; 26.2% had diabetes. Thirty-four per cent of the patients had not had a diagnosis of leg ulceration. Saline was the preferred cleansing solution. Compression therapy was used on all types of ulcer. Pain was a problem for 52.9% of the patients, of whom 9% did not receive analgesia. CONCLUSION: Nurses and doctors working in this community need to update their knowledge of wound treatments. PMID- 14533236 TI - Necrotising fasciitis: its aetiology, diagnosis and management. PMID- 14533237 TI - Rewarding best practice: the 2004 Journal of Wound Care/Tendra Honours and Scholarships. PMID- 14533238 TI - An in vitro analysis of the antimicrobial properties of 10 silver-containing dressings. PMID- 14533239 TI - Does lymphoedema bandaging reduce the risk of toe ulceration? PMID- 14533240 TI - Use of Allevyn heel in the management of heel ulcers. AB - OBJECTIVE: This small uncontrolled study examined the effectiveness of Allevyn Heel. METHOD: Twenty-two patients were recruited: 13 with heel pressure ulcers, four with diabetic heel ulcers, four with arterial heel ulcers and one 'other'. Ulcers were assessed before treatment and after the start of the dressing regimen at weeks two and four and at the end of the study. RESULTS: Patients were treated with Allevyn Heel for a mean of 47.2 days, and dressings were left in place for an average of 2.2 days before being changed. By the end of treatment, 32% of the ulcers had completely or almost healed, and a further 27% were showing evidence of granulation. Application was considered easy in 98% of instances, and 91% of the patients reported that Allevyn Heel was comfortable to wear. CONCLUSION: Allevyn Heel was found to be effective and easy to use. This extends the range of wounds that are suitable for management with Allevyn, although a larger-scale study is needed to confirm the utility of Allevyn Heel for these type of heel wounds. DECLARATION OF INTEREST: This study was supported by Smith and Nephew, Germany. PMID- 14533241 TI - Silver-containing dressings: have we got the full picture? PMID- 14533242 TI - Malodorous wounds--taking the patient's perspective into account. PMID- 14533243 TI - Mix and match. PMID- 14533244 TI - A foot in the door. PMID- 14533245 TI - Nothing to sneeze at. PMID- 14533246 TI - One for all. PMID- 14533247 TI - Experience of a lifetime. PMID- 14533248 TI - Give and take. PMID- 14533249 TI - Menorrhagia: endometrial ablation or hysterectomy? AB - BACKGROUND: For many woman who suffer from menorrhagia, treatment would previously have meant hysterectomy. Today there are a number of less invasive surgical options available, such as endometrial ablation, which involves removal of the endometrium and part of the myometrium. This article compares two endometrial ablative techniques--transcervical resection of the endometrium and thermal balloon therapy--with hysterectomy in the management of menorrhagia. CONCLUSION: While satisfaction with outcome is generally higher in women undergoing hysterectomy, satisfaction is also high following ablation and the procedure is less invasive and safer. Further research is required, however, to evaluate the long-term efficacy of ablation when compared to hysterectomy. PMID- 14533250 TI - Using non-invasive ventilation in acute wards: Part 1. AB - Non-invasive ventilation (NIV) is increasingly being used in domestic and acute health settings. Part one of this article identifies the benefits of NIV and describes the use of continuous positive airway pressure (CPAP). Part two, published in next week's Nursing Standard, discusses bilevel NIV, which has fewer complications and so is largely replacing CPAP. Part two also examines how to monitor and assess patients receiving NIV. PMID- 14533251 TI - Developing portfolios for peri-operative nurses. AB - Compiling and maintaining a professional portfolio is a requirement for all practising nurses. This article focuses on the process of building a portfolio for nurses working in operating theatres. PMID- 14533253 TI - The cultural virus. PMID- 14533252 TI - Lead the way. PMID- 14533254 TI - "Laser root canal therapy". PMID- 14533255 TI - Is your patient having a stroke? PMID- 14533257 TI - The aesthetic hands-on continuum. PMID- 14533256 TI - Implant treatment and the role of platelet rich plasma. AB - Implant supported restorations have become and continue to be a very popular clinical option for patients who are either partially or completely edentulous. This article, the third in a series dealing with implant dentistry, describes the role of platelet rich plasma as an adjunct to help with the healing process following tooth extraction, socket preservation, and, ultimately, replacement with implant supported restorations. A clinical case is used to illustrate surgical options available in the restoration of the posterior partially edentulous maxilla. PMID- 14533258 TI - New harvesting technique for bone grafting. Case reports. AB - BACKGROUND: Indications for grafting bone to the oral and facial skeleton are increasing as greater numbers of reconstructive procedures are performed. This article discusses a simple harvesting technique for autogenous bone. CASE DESCRIPTION: Successful outcomes of three grafted mal-union mandibular fractures and a grafted bilateral sinus lift procedure are reported. CLINICAL IMPLICATIONS: This system allows for simple, rapid retrieval of autogenous bone in multiple surgical procedures requiring bone grafts, and can be performed by all dentists and dental specialists. PMID- 14533259 TI - The success of organized dentistry begins at the component. PMID- 14533260 TI - Victoria's story. PMID- 14533261 TI - Wyeth found guilty of breaching infant formula advertising ban. 'Cynical and deliberate' breach of regulations by company. PMID- 14533262 TI - Government fails to deliver childbirth reforms, say MPs. Women still not given greater choice. PMID- 14533263 TI - Midwife struck off. PMID- 14533264 TI - Preventing premature births. PMID- 14533265 TI - Being used? Motives for user involvement. PMID- 14533266 TI - Building bridges: involving Pakistani women. PMID- 14533267 TI - Powerful sharing? Creating effective user groups. PMID- 14533268 TI - Culture, control and the birth environment. AB - These findings illustrate that women's needs are not being adequately met in many hospital birth units. Women, particularly those expecting their first baby, often know little about how the culture of hospitals varies, or the helpfulness of facilities and opportunities for control and one-to-one support that are more readily available at home or in a midwife-led unit. Nor do they know how much their opportunities for comfort and control may be compromised in a conventional hospital setting. If they are feeling anxious about whether they will be able to cope with the pain of labour and whether their baby will be born safely, it is perhaps not surprising that a significant proportion feel it is important to have access to an epidural service and a special care baby unit (House of Commons Health Committee 2003a). However, these facilities are not in themselves more likely to make labour straightforward and manageable. Midwives and organisations such as the NCT have a key role to play in sharing knowledge about what women actually find useful--or disruptive and unhelpful--during labour, so that all pregnant women can make choices that are informed by the full range of relevant information. The recommendations from the early 1990s, that women should have care from a known midwife, has not been realised consistently, although in environments that are highly medicalized neither knowing your midwife nor one-to one support seem sufficient to affect labour outcomes substantially (Johanson et al 2002). Where there is strong midwifery leadership, a clear philosophy of normality and one-to-one support, outcomes are different (Biringer, Davies. Nimrod et al 2000). Women appear to be offered more choices than a decade ago, but the range of options available still tends to be dominated by a medical model of care. Women still do not receive adequate information on the significance of alternatives as good-quality, evidence-based information is not [table: see text] consistently available, nor are they given the support to choose freely from the full range of options (Singh and Newburn 2000). However, evidence-based information leaflets alone are known to be inadequate to overcome a range of cultural barriers (O'Cathain et al 2002; Stapleton et al 2002). A small and growing proportion of women are having home births and have access to a midwife led unit. Use of a birthpool in labour has become more accepted in all birth settings, although access to suitable facilities and protocols for use in hospital units are sometimes restrictive. Further change is needed to provide care during labour as part of a midwifery model, so that the kinds of medical interventions women find intrusive can be limited as far as possible without compromising safety. PMID- 14533269 TI - The new pathology of shortness. PMID- 14533270 TI - The ethics of extreme prematurity and late abortion. PMID- 14533271 TI - Trust me--I'm a midwife. PMID- 14533272 TI - Going backwards: the concept of 'pasmo'. PMID- 14533274 TI - Genetic and environmental risk factors for the development of atopy and related diseases. PMID- 14533273 TI - Respiratory hazards in agriculture. PMID- 14533275 TI - The psychological impact of comprehensive pulmonary rehabilitation: which evidence? PMID- 14533276 TI - Guidelines for COPD management. PMID- 14533277 TI - Opto-electronic plethysmography. AB - Recent developments of Opto-Electronic plethysmography allow a noninvasive and accurate measurement of the volume of the chest wall and its different thoraco abdominal compartments, without any calibration specific on the subject under analysis. OEP provides the complete ventilatory pattern without using any device attached to the subject in any conditions and posture. Combined with pressure and airflow measurements, it can be used to study statics, dynamics and energetics of the chest wall and to estimate possible blood shifts between the thorax and the extremities. PMID- 14533278 TI - Pulmonary gene therapy. Realistic hope for the future, or false dawn in the promised land? AB - Pulmonary gene therapy offers the hope of treatment for conditions such as cystic fibrosis, lung cancer, pulmonary fibrosis and acute respiratory distress syndrome for which current therapy is inadequate. Although initial clinical trials in cystic fibrosis and non-small cell lung cancer have shown promise the results have not been as good as might have been anticipated. However, clinical improvement has been demonstrated in conditions such as haemophilia [82], cardiovascular disease [83], head and neck cancer [84] and X-linked severe combined immunodeficiency disease [85]. The lack of success of pulmonary gene therapy is due, in part on the physical barriers to transfection perfected by the lung to prevent toxicity from inhaled particles, and partly due to the poor transfection efficiency of non-viral systems, and the immunogenicity of viral systems, of gene transfer. The LID vector goes some way to addressing the problems associated with current gene delivery strategies. With continued improvements in the properties of both viral and non-viral gene delivery systems leading to improved transfection efficiency with reduced toxicity, as well as the development of strategies aimed at reducing the physical barriers to pulmonary transfection, and targeting gene delivery systems to the site of injury, it is likely that pulmonary gene therapy will be used successfully to ameliorate a number of devastating pulmonary conditions. PMID- 14533279 TI - Anti-IgE monoclonal antibody: a new approach to the treatment of allergic respiratory diseases. AB - The immunoglobulin E (IgE) antibody plays a central role in the allergic immune responses. The ability to reduce circulating IgE with a humanized monoclonal antibody (omalizumab) represents a new therapeutic approach for the treatment of IgE-mediated allergic diseases. The use of an anti-IgE antibody in the treatment of asthma was first suggested in preliminary studies in which omalizumab demonstrated efficacy in attenuating both the early- and late-phase bronchial responses to inhaled aeroallergens. Therapy with omalizumab has demonstrated both a significant beneficial effect on a number of measures and a favorable safety profile. It reduces the frequency of asthma exacerbations and the need for inhaled corticosteroids (ICSs), and improves asthma symptoms, lung function, and quality of life. The anti-IgE approach to asthma treatment has several potential advantages, such as the treatment of other concomitant atopic diseases (allergic conjunctivitis and rhinitis, atopic dermatitis, and food allergy) regardless of atopiC of allergic sensitization. PMID- 14533280 TI - How signaling pathways interact with gene transcription. AB - Activation of inflammatory cells by extracellular stimuli such as cytokines, viruses, allergens or oxidants lead to stimulation of intracellular amplifying signalling pathways. These activation pathways lead to enhanced inflammatory gene expression through activation of transcription factors such as nuclear factor kappa B (NF-kappa B). Upon activation, NF-kappa B translocates to the nucleus where it binds specific sites in the promoter regions of responsive genes in a time-dependent manner. This leads to the alterations in the structure of the surrounding DNA/chromatin complex resulting in local unwinding of DNA. Changes in chromatin structure occur as a result of recruitment of large, essentially pre formed, complexes that contain chromatin remodelling factors and histone acetyltransferases (HATs) such as CREB binding protein (CBP). Acetylations of histones are regulated by HATs whose activity is controlled by a number of factors including phosphorylation. Phosphorylation of HATs by signalling pathways, differentiate themselves from those controlling specific transcription factor activation and result in enhanced transcriptional activity as a result of cross-talk between signalling pathways at the level of chromatin modifications. PMID- 14533281 TI - Early life exposures and the prevalence of atopic disorders in a sample of school age infants. AB - This study was undertaken to determine the relationship between genetic factors, early life environmental exposures, and the prevalence of atopic disorders in infants. 402 children attending schools in rural Essex were sampled in June 1999. Logistic regression models were fitted to examine the association between asthma, eczema, and rhinitis prevalence and a range of genetic and environmental risk factors. After adjustment, infant asthma was associated with the report of a serious chest infection before 3 years of age, a parent employed in a professional occupation, an asthmatic mother, and an asthmatic sibling. Amongst infants who had attended nursery school there was also an affect of age at first attendance. Associations with a chest infection and a family history of atopic conditions were similarly strong predictors of eczema and rhinitis prevalence. The results suggest that genetic influences and patterns of exposure to infections appear to be the greatest influences on the development of infant atopy. PMID- 14533282 TI - Clinical benefits of a combined hospital and home-based exercise programme over 18 months in patients with severe COPD. AB - BACKGROUND: Long-term exercise training is capable of improving exercise performance and quality of life in patients with severe COPD. In the present study we examined the effects of an 18-month home-based training on the rate of hospital admissions and bronchodilator use as primary end-points. Secondary end points were exercise capacity and quality of life. METHODS: The study comprised 26 patients with severe COPD (20m/6f; mean +/- SD FEV1, 37 +/- 6% pred) who were recruited in a previous trial and randomised into a training (n = 14) and a control group (n = 12). After initial recovery from an exacerbation the training group had performed a 10-day walking training in the hospital. This was followed by 18 months of individually defined, supervised training at home that was integrated into the patients' daily activities. The control group had no exercise programme, neither in hospital nor at home. RESULTS: During the 18-month period patients of the training group showed a lower number of hospital admissions (total, n = 3 vs n = 14, p = 0.026; disease-related, n = 3 vs n = 12, p = 0.050) and used less short-acting beta 2-agonists (mean [95% CI], 2.4 [1.4-3.4] vs 5.7 [4.2-7.2] puffs per day; p < 0.001) than the control group. Furthermore, the improvements in 6-min treadmill distance and quality of life (CRQ) achieved in the hospital were fully maintained in the training group, whereas the control group did not show significant improvements at any time but a tendency toward deterioration. CONCLUSIONS: Our data indicate that an individually defined, home based, long-term walking programme initiated by a short hospital-based training can reduce disease-related medical consumption, in addition to sustained benefits in exercise performance and quality of life. PMID- 14533283 TI - Risk factors for chronic respiratory disorders in a sample of farmers in middle Italy. AB - OBJECTIVE: To evaluate the prevalence of respiratory symptoms in agricultural workers in a rural area of Tuscany and to relate them to some occupational risk factors. SUBJECTS AND METHODS: A cross sectional study was performed on a sample of 461 agricultural workers from two rural area of Tuscany. All subjects underwent to respiratory questionnaire, pulmonary function test and skin prick tests. Occupational risk factors were evaluated as response to questions related to 11 specific jobs, most of them related to cow-shed works. RESULTS: Subjects had high mean age and a long work duration. Smoking habit was significantly associated with symptoms of chronic bronchitis, while atopy was significantly associated with chronic cough, phlegm and dyspnea. In comparison with a control group of workers which did not report any occupational risk factors (reference group), logistic analysis showed that most symptoms were significantly associated with some specific jobs related to animal breeding, taking into account differences due to age, smoking habit, atopy and district. Mean pulmonary function tests were in the normal range, without any significant relationship with different risk factors, except smoking habit. Increased work duration was associated with significant decrease of pulmonary function. CONCLUSION: Symptoms of chronic bronchitis were frequent in this population of farmers of Middle Italy. The analysis of risk factors confirms a relevant role for some specific occupational risk factors, related to jobs in the cow-shed, in the development of respiratory disorders. PMID- 14533284 TI - Impact of comprehensive pulmonary rehabilitation on anxiety and depression in hospitalized COPD patients. AB - To prospectively evaluate the effect of inpatient pulmonary rehabilitation (iPR) on anxiety and depression as outcome measures in patients with COPD, we studied 149 consecutive adults COPD referred to our iPR after an exacerbation. Patients were divided according to the GOLD staging into: Group 1 (stage 2a, n = 48, FEV1 63 +/- 9% pred.), Group 2 (stage 2b, n = 53, FEV1 42 +/- 6% pred.) and Group 3 (stage 3, n = 48, FEV1 25 +/- 7% pred.). The iPR consisted of twelve 3-hours daily sessions. Hospital Anxiety Depression (HAD) Scale as well as 6-minute walk (6MWD) with evaluation of dyspnea (D) and leg fatigue (F) at rest and end of effort, and health related quality of life by means of St. George Respiratory Questionnaire (SGRQ) were assessed before (T0) and after (T1) the iPR. 6MWD, D and F at end of effort and SGRQ total score similarly improved (p < 0.001) in all groups after iPR. The mean level of HAD-anxiety (from 9.1 +/- 4.0 to 7.7 +/- 3.5, from 9.0 +/- 4.6 to 7.2 +/- 4.6 and from 8.1 +/- 4.1 to 6.7 +/- 4.3 in group 1,2 and 3 respectively) and HAD-depression (from 9.4 +/- 3.5 to 8.2 +/- 3.5, from 9.1 +/- 4.2 to 8.2 +/- 4.5 and from 9.0 +/- 4.0 to 7.4 +/- 4.5 respectively) similarly changed (p < 0.0001) over time in all groups. The total percentage of patients with abnormal score (> 10) of HAD-anxiety (from 31% to 21%) and HAD depression (from 30% to 22%) significantly decreased (p < 0.05) after the iPR. Inpatient pulmonary rehabilitation may improve levels of anxiety and depression as well as symptoms, exercise capacity and health related quality of life in moderate to severe COPD patients after an acute exacerbation. PMID- 14533285 TI - Management of COPD according to guidelines. A national survey among Belgian physicians. AB - Current management of COPD by Belgian physicians was compared with the recommendations of the recently published GOLD guidelines. A random sample of 386 general practitioners and 86 pulmonologists filled in a questionnaire based on the GOLD guidelines and examining their attitudes towards COPD management. Several important deviations from the guidelines were noted. Only few GP's performed spirometry themselves and about 55% of the diagnoses were not based on spirometry. Both GP's and pulmonologists used inhaled corticosteroids considerably more often than prescribed by the guidelines, with 49% and 25% respectively, prescribing them to all COPD patients. Chronic systemic steroids were also overused in stable disease, with 55% of the GP's and 52% of the pulmonologists prescribing them in patients with repeated exacerbations. GP's did not use enough systemic corticosteroids and overused antibiotics in the treatment of exacerbations. Pulmonologists did not pay enough attention to pulmonary rehabilitation, as only 44% of them had a structured programme. Pulmonologists did not use non-invasive ventilation frequently enough in the treatment of exacerbations, as only 22% would use it in the correct indication. Both GP's and pulmonologists did not prescribe enough pharmacotherapy for smoking cessation, with 35% and 46%, respectively using it. Some interesting differences between Dutch and French speaking physicians were noted. These specific deviations from the guidelines will be addressed in a second phase implementation project. PMID- 14533286 TI - Adult croup--a case report. AB - A case of a female adult patient with croup is described. Inflammation, as well as pseudomembranes, restricting the patency of trachea, developed within several hours after the first symptoms of infection were observed. Tracheostomy was performed Bronchofiberoscopies were repeated on a regular basis several times a day over a period of two weeks, with the removal of fibrinous casts and dense secretion being the only way to save the patient's life. PMID- 14533287 TI - Common variable immunodeficiency in an adult with recurrent pneumonia. AB - Primary immunodeficiency syndromes are rarely diagnosed among adults. Common variable immunodeficiency (CVID) is a congenital immunological disorder characterized by defective antibody production. In this report, we describe a 35 year-old male suffering from a common variable immunodeficiency, referred to us because of a lobar pneumonia. He had a history of recurrent pulmonary infections, which was present months before presentation, suggesting hypogammaglobulinemia. We found a severe hypogammaglobulinemia, which confirmed the diagnosis of CVID. His immunoglobulin profiles upon admission before infusion of immunoglobulin (normal ranges) were: IgG < 1.41 (8-17) g/l, IgA 0.25 (0.85-4.9) g/l, IgM 0.182 (0.5-3.7) g/l, and IgE < 2 (< 120) IU/ml. His HLA profiles were HLA A2 A26, B18 B38, Cw7, DR11 and DQ7 DQ9. He was treated with intravenous immunoglobulin. After this regimen, his IgG was maintained at > 6.0 g/L. On follow up, he has been free of opportunistic infections. In conclusion, CVID should be considered in the differential diagnosis of recurrent pneumonia in adults. PMID- 14533288 TI - Cardiac arrest due to ventricular fibrillation as a complication occurRing during rigid bronchoscopic laser therapy. AB - Laser bronchoscopy is a usually well tolerated procedure for the treatment of obstructive lesions on the tracheobronchial tree, with a very low morbidity and mortality rate. Cardiovascular complications, including atrial and ventricular arrhythmias, and myocardial ischemia, have only rarely been reported during laser bronchoscopy. Cardiac arrhythmias during such a procedure are usually well tolerated but occasionally may be life threatening. Here we report a case of a young, female patient affected by Pulmonary Tuberculosis with a cicatricial stenosis of the left main bronchus who developed an episode of prolonged cardiac arrest due to ventricular fibrillation (and no signs of acute myocardial ischemia) during rigid broncoscopic laser-therapy. Underlying Coronary Artery Disease and other cardiac abnormalities were also excluded by subsequent cardiovascular examination. The clinical implications are also discussed. PMID- 14533289 TI - Time to first cigarette; the best single indicator of tobacco dependence? PMID- 14533290 TI - Editorial board's eye view. PMID- 14533291 TI - Asking too much. PMID- 14533292 TI - Turning lives around. PMID- 14533293 TI - Mental health triage. PMID- 14533294 TI - When the roof caves in. AB - Rescuing patients after structural collapse can be alien to emergency nurses but their ability to adapt should stand them well to provide good patient care in these unusual circumstances. Generally, they are held back until patients are released from the collapsed structures or specific requests for analgesia, anaesthesia or aggressive fluid resuscitation are made. PMID- 14533295 TI - Detecting high-risk patients with chest pain. AB - AIM: To assess the ability of nurses using the Manchester Triage System (MTS) to identify those patients with chest pain requiring immediate electrocardiogram (ECG) and physician assessment within ten minutes. METHOD: A four-week prospective cohort of all patients attending with chest pain compared detection of risk by nurses using the MTS to that of researchers using best available evidence-based prognostic indicators from history. RESULTS: The study of 167 patients showed that nurses using MTS had a sensitivity of 86.8 per cent (95 per cent confidence interval (CI), 78.4-92.3 per cent) and a specificity of 72.4 per cent (95 per cent CI, 61.4-81.2 per cent) when identifying high risk cardiac chest pain. CONCLUSION: Nurses using the MTS are a sensitive tool for identifying high risk cardiac chest pain but further work is required to assess whether additional training can improve sensitivity. PMID- 14533296 TI - Emergency care. Advanced practice nursing in the US: an overview. AB - The CNS role preceded that of the NP in the United States. While CNSs have always been educated to master's degree level, NP education has been more varied although it is now predominantly at this level too. An APN with a master's degree in nursing can attend a post-master's degree certificate programme to obtain education in a new specialty area or to change roles. Emergency care is one clinical specialty area in which NPs and CNSs can obtain education specific to their area of practice. To function in the APN role, nurses must be licensed as registered nurses, have graduated from a NP or CNS educational programme and then have obtained state approval. Certification in the specialty may be required to obtain state approval. Nurse practitioners in emergency care have a scope and standards of practice although these are currently under development for CNSs in emergency care. Nurse practitioners function in roles where they provide direct patients care. Since the CNS role focus is improving nursing care, these individuals typically provide direct care to patients with complex health problems but also focus on care systems. PMID- 14533297 TI - Assessing acute myocardial infarction. PMID- 14533298 TI - Wernicke's encephalopathy. PMID- 14533299 TI - Nurse led fast track for vulnerable older people. PMID- 14533300 TI - Sharing the vision--report on the June 2003 leadership council meeting. PMID- 14533301 TI - AHNA Holistic Nurse of the Year 2003--Carla Mariano EdD, RN, HNC. PMID- 14533302 TI - Update on nurses practicing holistic therapies in Minnesota. PMID- 14533303 TI - Field trips for the kids. PMID- 14533304 TI - Weakening the grip of cluster headache. PMID- 14533305 TI - Travel needn't mean hassle when paying your bills. PMID- 14533306 TI - Travel essentials. PMID- 14533308 TI - A day in the life ... of a medical salesman. PMID- 14533309 TI - Reforming emergency care. PMID- 14533310 TI - The dilemma of conjoined twins. PMID- 14533311 TI - Managing a national pilot site project. The Derby Theatre Project experience. AB - The National Theatre Project was set up in March 2001 by the Modernization Agency to improve the patient and carer experience, improve employee satisfaction, optimise theatre utilisation and reduce cancelled operations. This is the first in a series of four articles, looking at issues involved in managing a national project, improving operating theatre performance and how to engage the organisation in innovation and change. PMID- 14533312 TI - Student nurse theatre placements. The new curriculum. AB - Following the implementation of the Fitness for Practice (FFP) nurse education curriculum, opportunities are available for student nurses to return to the operating theatre environment during their surgical module. This follows a ten year absence in most parts of Wales. This article investigates what realistic placement objectives are available for student nurses in Wales and how close mentor supervision can enhance the learning process and promote the clinical experience of theatre nursing. PMID- 14533313 TI - The theatre nurse role. A review of the literature. PMID- 14533314 TI - Role perception in a climate of change. The role of the anaesthetic nurse. PMID- 14533315 TI - Will fast-track units benefit nurses? PMID- 14533316 TI - Forcing people to give up drugs is not the answer. PMID- 14533317 TI - Nutrition. The big challenge. PMID- 14533318 TI - Community pharmacy and NHS Direct in new team initiative. AB - Over the past five years, NHS Direct has handled over 18 million calls and become one of the world's principal providers of telephone-based health care advice. Research has shown that almost six per cent of callers seek advice about medicines and approximately 40 per cent of responses include advice about medicines. It was as a result of this identified need that the community pharmacy project was born. It has since been rolled out across all NHS Direct sites. The project aims to provide a framework to enable NHS Direct advisers to offer safe and appropriate information and referral. PMID- 14533319 TI - What you need to know about ... sinusitis. PMID- 14533320 TI - Barium swallow. PMID- 14533321 TI - Understanding the physiological effects of unrelieved pain. AB - Pain produces a physiological stress response that includes increased heart and breathing rates to facilitate the increasing demands of oxygen and other nutrients to vital organs. Failure to relieve pain produces a prolonged stress state, which can result in harmful multisystem effects. Good acute pain management is an essential part of holistic nursing care. PMID- 14533322 TI - The development of a nurse-led outpatient orthopaedic clinic. AB - In the past 18 months nurses at the orthopaedic outpatients department at Harrogate District Hospital have radically transformed their roles by developing a nurse-led joint replacement follow-up clinic. This clinic, initiated in response to both local and national developments, has led to increased nurse autonomy as well as a demonstrable improvement in patient services. PMID- 14533323 TI - Laxatives should be the last resort in constipation. PMID- 14533324 TI - A world first for nursing. Interview by David Crouch. PMID- 14533325 TI - Equip as to care. Playing the charity lottery. PMID- 14533326 TI - Achieving safe sharps practice. AB - The use of sharps should be avoided where possible. However, when their use is essential, particular care is required in handling and disposal and, if possible, safer devices should be used. PMID- 14533327 TI - Risks and effects of sharps injuries. AB - Nurses face the risk of exposure to blood-borne infections if they suffer a needlestick or other injury from sharp devices such as lancets, scalpels and razors. In order to minimise their risk of contracting viruses such as HIV, hepatitis B and C, or other infections, risk assessment exercises must be undertaken so that safer systems of work can be implemented to protect nurses and other health care professionals. PMID- 14533328 TI - Gaining patient consent for blood tests following sharps injuries. AB - There is a high risk of staff receiving needlestick and other sharps injuries while working in health care. Concern is so great that a seminar on sharps injury prevention and single-use medical devices was held in March 2003 at the European Parliament in Brussels. Injuries caused by needles and other sharp medical devices, and the related risk of potentially fatal disease transmission, remain a major threat to the health and safety of health care workers across the European Union. PMID- 14533329 TI - A service to address the sexual health needs of the homeless population. AB - Between 1991 and 2001, the clinical workload of genitourinary medicine services increased by 155 per cent, and diagnoses of sexually transmitted infections (STIs) increased by 61 per cent in England, Wales and Northern Ireland (Public Health Laboratory Service et al, 2002). However there is a paucity of data regarding the rate of sexually transmitted infections in the homeless population. PMID- 14533330 TI - Health care-associated infections in primary and community care. AB - The National Institute for Clinical Excellence recently published guidelines for preventing health care-associated infections (HAIs) in primary and community care (NICE, 2003). These were commissioned by the Department of Health and NICE to complement previously published guidelines for preventing HAIs in hospitals (Pratt et al, 2001). The new guidelines aim to facilitate a seamless approach to using the best evidence for infection prevention practices as patients transfer between primary and secondary health care services. This article discusses the need for the guidelines and describes the methods used to develop them. PMID- 14533331 TI - Understanding the implications of NICE guidance on infection control. AB - Following the publication of Getting Ahead of the Curve (Department of Health, 2001), greater emphasis has been placed on infection control in primary and community care (called community care throughout this article). For years, guidance on reducing the incidence of preventable health care-associated infections in community care has been based on standards developed for acute care settings. But in many cases, practices common in acute care are not directly transferable to the community. Also, shorter hospital stays mean care previously provided in hospital is increasingly carried out in the community. PMID- 14533332 TI - Health implications of body piercing and tattooing: a literature review. AB - Tattooing and body piercing have become increasingly popular in recent years, particularly among young adults and adolescents. Montgomery and Parks (2001) say tattoos represent attributes such as individualism, risk, sexual fantasies and uniqueness, which are important for adolescents attempting to prove their individuality. PMID- 14533333 TI - A return of real leadership to the wards. PMID- 14533334 TI - Incorporation of the School of Oral Health Sciences of the University of Stellenbosch and the Faculty of Dentistry at the University of the Western Cape. PMID- 14533335 TI - R Lalloo and the perverse incentives. PMID- 14533336 TI - Effectiveness of four manual toothbrushes in a cohort of patients undergoing fixed orthodontic treatment in an academic training hospital. AB - A single-blind, cross-over study design was used to evaluate the effectiveness of four different manual toothbrushes and to determine whether patient toothbrush preference is directly related to plaque control in patients with fixed orthodontic appliances. The brushes evaluated were Orthodontic Oral-B, Oral-B Advantage 30, Colgate Precision and Aquafresh. Forty-six subjects, aged 11 to 27, undergoing fixed orthodontic treatment were screened and recruited with parental consent. Subjects were randomly allocated into four groups. All subjects had their teeth scaled and polished at week 0. Baseline recordings of PI and GI were done four weeks later and the first toothbrush was given. After using the toothbrush for a period of 2 weeks the PI and GI were again recorded and teeth were again scaled and polished. A period of four weeks elapsed before new baseline recordings were done and the sequence followed as described for the first toothbrush. This was done until all subjects had used all four toothbrushes. At the end of the clinical trial, each subject was asked which toothbrushes they preferred. PI and GI values were relatively low at baseline as well as after the use of the toothbrushes. General linear model procedure showed no statistical difference between the Mean Plaque Index (MPI) before and after use of each toothbrush as well as the Difference in Mean Plaque Index (DMPI). There was a slight difference in the Difference in Mean Gingival Index (DMGI) between the Colgate Precision and Aquafresh toothbrush. For all the other comparisons general linear model procedure showed no difference between the Mean Gingival Index before and after use of each brush. There was no correlation between the toothbrush preferred by the patient and oral cleanliness as measured by DMPI and DMGI. PMID- 14533337 TI - Cancrum oris--reflected. PMID- 14533338 TI - The Umgeni bridge, a resin bonded pontic using a preformed fibre post an aesthetic procedure of low biological and fiscal cost. AB - A missing anterior tooth or open embrasures destroys the aesthetics of a smile. The replacement of a missing anterior tooth is one of the most rewarding and challenging aspects of restorative dentistry. There are a number of techniques to do this, however, most involve the use of a technician, which means additional cost to the patient and multiple visits. In today's financial climate with practice expenses climbing and medical aid remuneration dwindling, one must seek methods to reduce costs and provide patients with more affordable dentistry. In addition, in crisis situations such as the traumatic loss of a tooth an immediate replacement technique albeit temporary is necessary. To replace a missing anterior tooth directly and immediately presents a rewarding challenge to the clinician. The Umgeni bridge named after the river that flows on the north side of Durban, is particularly useful in replacing congenitally missing laterals after completion of orthodontic treatment in the case of congenitally missing laterals and cleft palates. PMID- 14533339 TI - Inlay design--wrap around versus overlay design. PMID- 14533340 TI - Evidence-based dentistry in practice. PMID- 14533341 TI - General practitioner's radiology case 13. PMID- 14533342 TI - [The smile line, a literature search]. AB - Beautiful teeth, visible when smiling, are in line with the present ideal of beauty. The display of teeth when smiling is determined by the smile line: the projection of the lower border of the upper lip on the maxillary teeth when smiling. On the basis of a literature search the determining methods of the smile line are discussed, demographic data of the position of the smile line are given, and factors of influence are examined. There is no unequivocal method for determining the position of the smile line. A rough distinction can be made between qualitative and (semi)-quantitative methods. The (semi)-quantitative methods have clear advantages for research purposes, but their reliability is unknown. It was demonstrated that among minimally 40% of subjects the maxillary gingiva was not visible when smiling. The mandibular gingiva was not visible when smiling among more than 90% of subjects. Furthermore, it appeared that among women the smile line was on average higher situated than among men and that it has not yet been proven that the smile line will be situated lower when growing older. PMID- 14533343 TI - [Spontaneous blood blister formation swellings of the oral mucosa]. AB - An 88 year-old man was hospitalized because of bullous pemphigoid of the skin with detectable autoantibodies against type XVII collagen. During his stay in the hospital, he developed easily bleeding hemorrhagic bullous lesions of the oral mucosa, which were thought to be an oral manifestation of bullous pemphigoid. A thorough blood examination showed acquired haemophilia A, related to the development of autoantibodies against factor VIII. PMID- 14533344 TI - [Unilateral nasal bleeding due to an intranasal tooth]. AB - Intranasal teeth is a rare phenomenon. In order to understand it's etiology access to high-quality case reports is necessary. Given that intranasal teeth often induce morbidity through obstruction, rhinorrhoe or nasal bleeding, more rapid and effective diagnosis and treatment are needed. This article presents two case reports and a review of the literature, revealing that different methods for the removal of intranasal teeth are available. A new method is removing the tooth using endoscopy. It was suggested that this method reduces the risk of comorbidity associated with more conventional removal methods. PMID- 14533345 TI - [Sarcoidosis]. AB - Sarcoidosis is a chronic generalized benign granulomatous disorder of perhaps an autoimmune character which can afflict many organs. The insidious form prevails, a (sub)acute onset covers about one third of cases. In most patients the lungs are involved; tissue damage by granulomas and inflammatory cells causes a reduction of respiratory capacity, with dyspnoea as a result. Other often affected organs are larger joints, skin and eyes. The diagnosis is based on the symptoms, thoracic x-ray and histology. In most cases prednisone is the drug of first choice. It is tantalizing to have to decide whether to treat or not. The decision is based on the severity of the process in vital organs. PMID- 14533346 TI - Do all drugs have to be destroyed at the expiration date? PMID- 14533347 TI - Adverse response to pegylated interferon therapy in two patients with chronic hepatitis C. AB - Pegylated interferons have recently been approved for treatment of hepatitis C. The safety of these formulations is reported to be similar to that of non pegylated interferon. We present two patients who experienced exacerbations of their liver disease following administration of pegylated interferon alfa-2b. Vigilant monitoring of patients treated with these new agents is recommended. PMID- 14533348 TI - Descriptive epidemiological analysis of diving accidents in Hawaii from 1983 to 2001. AB - The Hyperbaric Treatment Center (HTC) at the University of Hawaii, has evaluated and treated over 1100 divers for dysbaric disease from 1983 to 2001. We describe some epidemiological parameters and compare trends between local residents and tourist divers in this article. Data obtained from this review were analyzed for age, gender, type of injury and resident status. While trends in Hawaii have mirrored national figures, we did determine that there were some significant differences between resident and tourist divers' patterns of injury over this period of time. PMID- 14533349 TI - Pelvic examination teaching: linking medical student professionalism and clinical competence. AB - The Professional Patient teaching program is an essential part of assuring competency in the performance of the pelvic examination and professional behaviors. Through a series of integrated teaching sessions from the first year reproductive anatomy laboratory, the second year basic clinical pelvic examination teaching program, to the third year teaching program, students perform at an enhanced level of clinical competency and professional behavior. A Professional Patient commented: "We create a safe environment where medical students not only learn the clinical portion of the exam, but also focus on the patient as the primary source of information on patient comfort. Students receive immediate feedback from us and have ample opportunity to ask questions about aspects of the clinical pelvic examination or doctor/patient communication skills. We guide them, teach them, and help them prepare for examinations with other patients who will not be as open or in tune with their bodies as we are." PMID- 14533350 TI - Laboratory analysis of HER-2/neu: search for the optimal testing strategy. PMID- 14533351 TI - [Focus on colorectal adenomas--therapeutic possibilities in precancerous changes and conditions of the colon and rectum in our daily practice]]. AB - The health government and also the professionals except the new screening test programme to put a stop the increasing number of colon and rectum cancer diseases, which have caused high mortality in Hungary. Believe in the success of this programme the authors give a short overview about the precancer status and alterations of the large bowel, focusing on colorectal adenoma. In particular, the pathological anatomy, symptomatology, diagnostics, therapy, treatment and prevention possibilities of adenomas are described in details. There were 3419 total colonoscopies carried out in the Laboratory of Endoscopy in Albert Schweitzer General Hospital of Hatvan between 31st January 1991 and 31st December 2001. All patients had bowel motility problems, abdominal pains, bloody stools or change of bowel movement habit. 941 large bowel polyps were diagnosed at 628 patient. 821 of them were removed with endoscopic polypectomy, 11 with mucosectomy, 43 with piecemeal polypectomy and 20 with surgical interventions. 38 carcinoma in situ and 94 serious dysplasia alteration were removed. After the endoscopical therapeutical interventions 5 complications were detected and one patient died. There were no complications after the surgical interventions. They analyse their patients in base of the polyps place, shape, number and histological structure of alterations, level of dysplasia and the type of interventions. It is confirmed that the total colonoscopy is the most adaptable method for examining the large bowel, and endoscopic polypectomy is the most efficient colorectal cancer preventing method, which could only be realized with the successful interdisciplinary cooperation of the gastroenterologist, the surgeon, the pathologist and radiologist. PMID- 14533352 TI - [Seroprevalence of Helicobacter pylori infection and anti-CagA positivity in the county Szabolcs-Szatmar-Bereg]. AB - The authors studied the seroprevalence of Helicobacter pylori infection and the risk factors for infection, among adult volunteers from Szabolcs-Szatmar-Bereg county, East Hungary in year 2000. Sera were collected from 756 adults (328 males, 428 females, m/f = 1.5:2; mean age 42.4 yrs.; range 18-69 yrs.). Anti Helicobacter pylori IgG and anti-CagA IgG antibodies were identified serologically using a sensitive ELISA technique. A questionnaire was completed to obtain necessary informations. The overall rate of seropositivity was 58.6% (45.2% in those aged 18-29 and 69.6% in those aged 50-59). Seropositivity was significantly higher in rural population than in urban area (p < 0.001), sharing a bed (p < 0.001), if parents were manual workers (p < 0.001), between keepers of cat (p < 0.005). The authors found a significant inverse correlation between Helicobacter pylori infection and educational level (p < 0.001), income (p < 0.005), absence of water supply and sanitation (childhood: p < 0.001, adulthood: p < 0.05). There were no significant differences in Helicobacter pylori infection related to gender, subject's occupation, smoking and drinking habits, gastric symptoms, family history of ulcer or gastric cancer. 67.5% of Helicobacter pylori positives were anti-CagA positives too. Helicobacter pylori positivity is higher than in developed countries, and than in the more developed county Vas. PMID- 14533353 TI - [Potential consequences of the sudden postnatal drop of estrogen levels in preterm neonates]. AB - Estrogens have profound effects on bone- and lipid homeostasis, modulate immune functions and play a major role in the regulation of the development and function of central nervous system, cardiovascular system and respiratory system. They induce hepatic protein production. There is a strong connection between estrogens and several hormone systems including growth hormone--IGF axis and renin angiotensin system. The extragenital effects of estrogens are probably more evident in the fetus than in fertile women. However, direct data are not available because of the lack of estrogen-deficient pregnancy models. Theoretically, in immature infants the premature cessation of in utero high estrogen levels is associated with the progression of perinatal acute complications. The volume of uterus in female premature infants is lower than in term newborns, the plasma levels of atherogen lipids are high, osteopenia due to inadequate bone acquisition is frequent. Inflammatory complications are common. Recently, the physiological effects of estrogen- and progesterone supplementation of premature infants have been evaluated in a pilot study. The preliminary results indicate, that the risk of bronchopulmonary dysplasia, osteopenia might be decreased in treated infants. Further studies are needed to confirm the effectivity and long term safety of estrogen supplementation of preterm neonates. PMID- 14533354 TI - [Physiologic mechanisms in extremity veins supporting chronic orthostatic tolerance]. AB - As a consequence of chronic orthostatic load of the organism several long-term antigravitation defense mechanism are activated in the lower extremity veins and microvessels. The following adaptive mechanisms are brought into focus: acute pressure-induced myogenic tone and its augmentation which promote the venous capacity autoregulation, potassium channels counterregulating the depolarization of the venous smooth muscle membrane, functional and structural remodeling of sympathetic innervation in the vascular wall, changes in venous network properties, decreased capillary supply in oxidative muscles of the extremities, and probably transendothelial homeostatic water transport in the wall of veins. PMID- 14533355 TI - [Current aspects in neurocysticercosis]. AB - Neurocysticercoses with intraventricular cysts are presented in two operated patients. Cysticercosis, the most frequent parasitic infection of the central nervous system develops only sporadically in Hungary. In this paper epidemiology, different presenting forms, diagnosis, differential diagnosis and therapy are summarized. The purpose of the authors with this publication is to call attention to the diagnosis and the adequate treatment of neurocysticercosis imported from endemic areas. PMID- 14533356 TI - [Treatment if acute hepatic failure]. PMID- 14533357 TI - [Eugenics in neurology and psychiatry in Hungary between the two World Wars]. AB - Eugenics became an organized movement in Hungary in 1914 with the establishment of the Section of Eugenics within the Association of Social Sciences (Tarsadalomtudomanyi Egyesulet). Its secretary, the prominent biologist Istvan Apathy jr., defined the aims and the place of eugenics within the sanitation as well as the necessary steps of organization which should be taken in Hungary. It was especially some Hungarian psychiatrists: Mark Goldberger, Lajos Namenyi, Gyula Donath and Laszlo Benedek, the Hungarian Representative of the Committee of International Eugenical Organisations who after World War I urged eugenical law which envisaged voluntary sterilization. According to the opponents of eliminative eugenics--like Karoly Csorsz, one of the first distinguished medical geneticists in Hungary--sterilization had not been scientifically based yet, because the ways and the probability of heredity of nervous and mental diseases, except for Huntington chorea, had not been explored yet. The outstanding neurologist, Karl Schaffer also opposed the sterilization bill worked out by Professor Benedek, so it was rejected by the National Council of Public Health in 1932 and was not discussed by the Hungarian Parliament either. In spite of its illegality, some sterilizations were executed, for example with the consent and the request of parents in the case of mentally retarded female to stop her reproduction. Only positive eugenics was legally supported by the marriage advisory bureau in the capital, but the interest in it was rather limited in interwar Hungary. PMID- 14533358 TI - Call of the wild. PMID- 14533359 TI - Toil and trouble. PMID- 14533360 TI - No kidding. PMID- 14533361 TI - The HSJ interview: Dr Sue Atkinson. Double impact. Interview by Helen Mooney. PMID- 14533362 TI - Clinical management--where medicine meets management. Trails of blood. Interview by Carol Lewis. PMID- 14533363 TI - Finance. Bond on the run. PMID- 14533364 TI - Health inequalities. On a roll. AB - The New Opportunities Fund is now five years old and has distributed 750 m Pounds, generally to address health inequalities. Healthy living centres have been a high-profile NOP beneficiary and are now being evaluated. Morecambe Bay suffers from a diverse range of inequality issues and has responded with a series of initiatives. PMID- 14533365 TI - HSJ people. Fit for purpose. PMID- 14533366 TI - Facilitation of clinician-initiated research. PMID- 14533367 TI - Cryotherapy after total knee replacement: a survey of current practice. AB - BACKGROUND AND PURPOSE: Cryotherapy is commonly used during physiotherapeutic rehabilitation after total knee replacement (TKR). Evidence for treatment effectiveness is contradictory and there are no clinical guidelines to inform treatment protocols within this patient group. The present survey investigated current cryotherapy practice after TKR throughout the United Kingdom (UK). METHOD: A postal survey, containing open and closed questions, was distributed to senior inpatient orthopaedic physiotherapists (n = 453). RESULTS: The response rate was 67% (304/453) and 58% (263/453) of the responses were completed by physiotherapists who treated TKR patients in the acute phase. Of these, 33% (85/263) routinely applied some form of cryotherapy after TKR. Physiotherapists working in the private sector were more likely to use cryotherapy and had greater access to Cryocuff equipment. The two main methods of cryotherapy application were the Cryocuff device, 59% (155/263) and crushed ice, 30% (79/263). Treatments were applied most frequently between 24 and 48 hours post-surgery, for 20 minutes, twice a day. Lack of proven efficacy was the most frequently stated reason for not applying cryotherapy treatment, and swelling was the most common indicator for treatment. There was particular uncertainty about the physical management of the Cryocuff device. CONCLUSIONS: There was little consensus with regard to treatment indicators, method of application and the management of cryotherapy after TKR. The results highlight a lack of consistency in the application of cryotherapy after TKR, indicating a need for further research. PMID- 14533368 TI - Impact of an exercise and walking protocol on quality of life for elderly people with OA of the knee. AB - BACKGROUND AND PURPOSE: The knee is the weight-bearing joint most commonly affected by osteoarthritis (OA). The symptoms of pain, morning stiffness of short duration and physical dysfunction in the activities of daily living (ADL) can have an effect on many aspects of health, affecting quality of life. Regular and moderate physical activity adapted to individuals' life-styles and education, and joint protection strategies have been advocated as conservative management. The purpose of the present study was to assess the impact of an exercise and walking protocol on the quality of life of elderly people with knee OA. METHOD: The study design was a randomized controlled clinical trial. The subjects comprised 50 elderly people, aged 65 or more, with knee OA who had been referred to the geriatric outpatient unit for rehabilitation. Changes in severity of pain and quality of life were compared between a control group (CG) and an experimental group (EG). Both groups participated in an educational session and the EG also received a 12-week exercise and walking protocol. Both groups were assessed at baseline and after three and six months by an independent observer. The Lequesne Index of Knee OA Severity (LI), the Health Assessment Questionnaire (HAQ) and the Medical Outcomes Short-Form Health Survey (SF-36) were used as measurement instruments. RESULTS: In the CG, the measures of quality of life (SF-36), the HAQ and the LI between subjects did not yield statistically significant differences over the three measurement points. For the EG, there was a significant improvement in function, measured by HAQ, and decreasing OA symptom severity, measured by LI. For the SF-36 there were significant improvements in physical function, functional role limitation and pain. Comparisons between the groups showed statistically significant differences after three and six months for all measures, except for the SF-36 emotional domains. CONCLUSION: The exercise protocol and walking programme had a positive effect on the quality of life of elderly individuals with knee OA. PMID- 14533369 TI - Age- and gender-related changes in the temporal-spatial characteristics of forwards and backwards gaits. AB - BACKGROUND AND PURPOSE: Backward walking is used increasingly as a rehabilitation technique for individuals with neurological and orthopaedic impairments. The purpose of the present study was to examine changes in the temporal-spatial characteristics of gain resulting from walking backwards as opposed to forwards, and to determine age and gender effects on these changes. METHODS: Thirty young and 40 aged, independently functioning, subjects were asked to walk forwards and backwards across a computer-based walkway system, providing data on gait velocity, stride length, cadence, swing phase and double support phase. Subjects were divided into groups based on age (young and old) and gender, and each subject was tested under two walking conditions (forwards and backwards). Five temporal-spatial gait parameters were evaluated separately as a function of the three independent variables, with the walking condition repeated for each subject. RESULTS: Backwards ambulation is characterized by a slower velocity, shorter stride length and an increased double support phase in both young and older adults. These changes were significantly greater in the older subjects, among whom the swing phase was also decreased. Cadence, however, was not affected by direction of ambulation in either group. The female subjects had a shorter stride length in both movement directions, associated with reduced speed only in backwards ambulation. CONCLUSIONS: Older individuals are capable of walking backwards for short distances. However, changes in gait characteristics typical to the reversal of movement direction are accentuated with age. These effects must be considered when planning to use backwards ambulation as a rehabilitation technique for older individuals. PMID- 14533370 TI - The effect of task uncertainty on muscle activation patterns in 8-10-year-old children. AB - BACKGROUND AND PURPOSE: Postural muscle activity accompanying voluntary arm movements has been researched extensively in adults; however, few studies of this kind have been conducted with children. It is not clear whether children respond like adults to internal perturbations, and if so, whether this response is similar across a variety of task conditions. The aims of the present study were to determine the postural muscle activity demonstrated by 8-10-year-old children when performing a rapid shoulder flexion movement and to compare the pattern produced under simple reaction-time versus complex reaction-time conditions. METHOD: Fifteen children with typical motor development participated in this comparative design study. The activity of anterior deltoid and contralateral rectus abdominis, internal oblique, erector spinae, rectus femoris, biceps femoris, tibialis anterior and gastrocnemius muscles was recorded via surface electromyography (EMG). Two tasks were performed: a simple task involving a right shoulder flexion movement; and a complex task involving a right or left shoulder flexion movement. Reaction time (RT) and movement time (MT) of the arm action were also recorded. RESULTS: Muscle latencies in the complex task were longer than those for the simple task, although only the gastrocnemius muscle showed a significant difference (p = 0.013). Reaction time was also significantly later in the complex task (p = 0.003). Movement time was not significantly different between conditions. CONCLUSIONS: A decrease in task certainty delayed postural muscle activation and reaction time in children aged 8-10 years. The study also demonstrated that children in this age group utilized a mixed postural strategy rather than a proximal-to-distal or distal-to-proximal pattern, reported in adult studies. Movement time remained unchanged. PMID- 14533371 TI - Effect of manual hyperinflation on haemodynamics in an animal model. AB - BACKGROUND AND PURPOSE: Manual hyperinflation is a physiotherapy technique that improves static compliance and mobilizes secretions, but has the potential to alter haemodynamic function. The aim of the present study was to investigate the effects of manual hyperinflation on haemodynamic function in a healthy animal model, without the usual confounding effects inherent in an heterogeneous intensive care population. METHOD: The study used a within-subjects design, in an animal research theatre. Nine healthy sheep (eight Border Leicester, one Merino, mean weight 39.5 kg, standard deviation (SD) 1.6 kg) completed the study. The sheep were induced (thiopentane 15-20 ml), intubated, ventilated and surgically instrumented for an arterial line and pulmonary artery catheter. Anaesthesia was maintained by 1.5% halothane/oxygen. Manual hyperinflation was delivered for two minutes with a Mapleson C circuit, using a peak inspiratory pressure of 35 cmH2O and an inspiratory:expiratory ratio of 2:1. RESULTS: Mean tidal volume during manual hyperinflation was 294% (SD 22%) of the ventilator tidal volume. A paired Student's t-test demonstrated that cardiac output (thermodilution method) decreased significantly (p < 0.05) and systemic vascular resistance increased significantly (p < 0.01) after manual hyperinflation. A repeated-measures analysis of variance (ANOVA) and a least-significant difference pairwise comparison revealed that mean arterial pressure and pulse pressure decreased significantly during (p < 0.01) and increased significantly (mean arterial pressure, p < 0.05 and mean pulse pressure p < 0.001) after the technique. Pulmonary artery pressure also increased significantly during manual hyperinflation (p < 0.01). There were no significant effects on right atrial pressure, pulmonary artery occlusion pressure or heart rate. CONCLUSION: Significant haemodynamic changes occurred in this animal model. The increased intrathoracic pressure, applied for an increased period during inspiration, decreased cardiac output with compensatory vasoconstriction evident by the increased systemic vascular resistance and mean arterial pressure. The results suggest that there may be a decrease in cardiac output after increased positive pressure in subjects with normal cardiac and respiratory function. PMID- 14533372 TI - [On validity of the term "chronic" rupture of abdominal aortic aneurysm]. AB - Treatment of patients with abdominal aortic aneurysms has been analyzed. From 506 patients (265 emergency and 241 elective cases) treated in the department of urent vascular surgery 36 had "chronic disruptions". Validity of this term is discussed. The authors consider the term "repeated disruption" as the most adequate. PMID- 14533373 TI - [Lateral extraperitoneal mini-approach for adrenalectomy]. AB - The techniques of adrenalectomy from lateral extraperitoneal mini-approach are described. This approach creates necessary space and ensures adrenalectomy in organic diseases of the adrenal glands. Fourteen operations with good short- and long-term results were performed. Limiting factors of this approach are the following: depth of the surgical wound 15 cm and more due to stage IV obesity and acute costal angle, sclerosis of retroperitoneal fat, significant splenomegaly, size of adrenal tumor more 6 cm. The "assisting hand" and endovideoscopy expand possibilities of the method. PMID- 14533374 TI - [Endoscopic drainage of postnecrotic pancreatic cysts]. AB - Results of endoscopic drainage of postnecrotic pancreatic cysts in 12 patients were analyzed. Possibilities of the method are limited by location of the cysts in pancreatic head and corpus. Clear visualization of the cysts on the gastric or duodenal wall is the main condition for this procedures feasibility. Surgery must be started with point burning in the wall that is safe and permits the surgeon to stop procedure when cyst is not found. When the cyst is drained through the duodenal lumen, it is necessary to visualize Veter's papilla and perform the procedure below it to prevent lesion of a distal part of the common bile duct. Section of the cyst with wall thickness to 3-4 mm from the duodenal lumen and the ikness to 6-7 mm from the stomach is safe enough. Endoscopic procedures are not indicated in hypertension in the bile ducts. PMID- 14533375 TI - [Antegrade papillosphincterotomy in laparoscopic cholecystectomy]. AB - Stenotic lesions of the distal part of the common bile duct are often revealed in laparoscopic cholecystectomy. From 1997 intraoperative antegrade papillosphincterotomy (APST) has been introduced in clinical practice of the endoscopic department of the A.V. Vishnevsky Institute of Surgery as the method able both to treat choledocholithiasis and restore adequate bile outflow. Indications to APST were stenosis of Vater's papilla (13 patients), stricture of a terminal part of the common bile duct (6 patients), choledocholithiasis in combination with distal bile duct stenosis (20 patients), isolated choledocholithiasis (4 patients). APST was successful in 36 patients and in 35 ones it was a final procedure for correction of ducts lesions. There were no complications of APST. It is an effective method in the treatment of intraoperatively defected stenotic lesions of the biliary tracts and choledocholithiasis. It makes the base of one-stage surgical policy in line with cholangioscope-assisted lithoextraction, intraoperative retrograde papillosphincterotomy and balloon dilatation of Vater's papilla. Antegrade papilloshincterotomy in patients with cholelithiasis and lesions of the ducts revealed during surgery permits one to reduce number of complications, time of hospital stay and increase comfort of surgical care. PMID- 14533376 TI - [Problems of hemostasis and hermetic properties in liver resections with application of fibrin-collagen substance]. AB - Forty-seven patients with various diseases of the liver were treated from 1992 to 2002. Patients had undergone liver resections of different scope. In 20 patients (study group) wound covering "Tachokomb" was used for final hemostasis and high hermetic properties of wound surface. Conventional suturing of the vessels and bile ducts was performed in 27 patients of the control group. Reliable hemostasis of the hepatic stump after anatomic resection was achieved in all the patients of the study group. Application of wound covering decreased time of final hemostasis, severity of hemorrhage and blood transfusion during surgery. Biliary complications after surgery were seen more often. Application of combined glue composition failed to reduce significantly rate of biliary complications after surgery, their rate increased proportionally to extension of liver resection. PMID- 14533377 TI - [Jejunostomy by Midle in patients with insufficiency of anastomotic sutures of upper parts of gastrointestinal tract]. AB - Insufficiency of anastomosis is one of the main causes of peritonitis. Progressed cachexia due to great losses through intestinal fistulas in combination with peritonitis was the indication to jejunostoma creation. Jejunostomy by Middle in the treatment of postoperative peritonitis due to sutures insufficiency of upper anastomosis of gastrointestinal tract permits to optimize treatment: to realize adequate enteral nutrition, intestinal decompression, to return fistula's losses. Polyorganic insufficiency, number of repeated surgeries and volume of loss through fistulas influence results of the treatment. Respiratory distress syndrome and errosive bleeding have the most prognostic significance among the syndromes of polyorganic insufficiency. PMID- 14533378 TI - [Healing of purulent wounds of soft tissues in local treatment]. AB - GOALS: To improve results of treatment of patients with purulent wounds of soft tissues of various etiology and location. METHODS: Developed method of local treatment was used in patients with purulent wounds of soft tissues. Efficacy of the treatment was evaluated with clinical, cytological, bacteriologic and bacterioscopic methods. RESULTS: When proposed antibacterial drugs were used, cleaning of wounds in inflammatory phase procedea 2 times faster compared to local treatment with standard drugs. Collagen wound covering Lioplast promoted in phases of regeneration and epithelisation 1.5 times faster reparative processes in the wound and prevention of secondary contamination of the wound. CONCLUSION: This method permits to improve results of treatment and to shorten hospital stay of patients. PMID- 14533379 TI - [Surgical treatment of a carbuncle]. AB - The results of the treatment of 250 patients with a carbuncle of various location were analyzed retrospectively. A differential approach to choice of surgical method with consideration of the degree, phase and localization of inflammation is preferable. Early and radical surgery, antimicrobic drugs, infusion therapy make base of up-to-date and adequate treatment. In the phase of infiltration deep incision often stops the disease or promotes a decrease of the necrotic zone. In suppurations linear incisions were made only on the face for cosmetic reasons. In other areas a few large connected T-, [symbol: see text]-, [symbol: see text]- or cross-formed incisions were preferable. In 30 cases I-formed incision with two rectangular flaps was used. PMID- 14533380 TI - [Results of an open comparative study on efficacy and safety of Voscopran bandages used for local treatment of venous ulcers in the II and III stages of wound healing]. PMID- 14533381 TI - [Phlegmon-necrotic rose]. PMID- 14533382 TI - [Mesothelial cyst of the diaphragm]. PMID- 14533383 TI - [Diagnosis of diaphragm damages]. PMID- 14533384 TI - [Duplication of the gall bladder]. PMID- 14533385 TI - [Phytobezoar of the stomach as the cause of intestinal obstruction]. PMID- 14533386 TI - [Occlusive intestinal obstruction due to gallstones]. PMID- 14533387 TI - [Surgical videothoracoscopy in children]. PMID- 14533388 TI - [Lasers in thoracic surgery]. PMID- 14533389 TI - [Surgical approaches to adrenal tumors]. PMID- 14533390 TI - [Problem of surgical suture of the colon]. PMID- 14533391 TI - [Professor Aleksei Sergeevich Rovnov]. PMID- 14533392 TI - The little puppet: working with autistic defences in mother-infant psychotherapy. AB - This paper outlines a constellation around birth in our society that is related to advanced western medicine and the way that families move around the globe. It describes a brief therapy with a three-year-old child who was diagnosed as autistic, and a mother who resorted to 'heroic' defences against the anxiety of being unsupported in a strange country. The therapist undertook to process the catastrophic experiences of the birth as expressed in the child's play and transform the accompanying monstrous fantasies into human experiences and thoughts. By providing a maternal container for both mother and child, the therapist was able to dissolve the couple's negative enmeshment and help create a space in the mother's mind in which she could process her own feelings, and thereby process and understand the child's feelings and communications. PMID- 14533393 TI - Possible psychosomatic hazards to the therapist: patients as self-objects. AB - This speculative paper concerns certain fundamentals of healing and psychotherapy which we mistakenly tend to take for granted. I discuss our need for the feeling of harmony, wholeness, and oneness. I call this archetypal need our 'normal autistic expectation'. When met, we experience well-being and 'healing'. If not sufficiently and reliably met, this expectation becomes an omnipotent demand ('autistic demand'). Frustration then brings about angry destructiveness, either outwardly directed or inwardly directed, with bodily changes which must be processed if bodily damage is to be minimized. Bereavement, the loss of a person necessary for one's feeling of wholeness (a 'self-object'), is an extreme and well-researched example of such damage. Our selfobjects are 'healing' when they help us to complete our sense of self. Our patients-, our profession, our colleagues, our place of work and our financial security are normally all part of our self-object structure. I give examples where patients' own needs for survival or intactness mean that they have to externalize their own hurt and anger for long periods of their therapy. This often means that the therapist's own wholeness and health are under attack, and even damaged permanently, or at least until the damage can be processed. The nature of 'processing' in this sense is therefore in need of energetic research. PMID- 14533394 TI - Understanding Jung: recent biographies and scholarship. AB - The tendency to associate Jung with Freud has undergone a change and both are increasingly perceived as founders of depth psychological schools whose exact relationship is unclear. The separation of the two was largely due to Jung's rejection by the psychoanalytic community because of his perceived spiritual inclinations. Recent scholarship has emphasized these spiritual inclinations in both a positive and negative way and brought to light Jung's non-Freudian sources, while other Jungian practitioners are seeking a closer association with psychoanalysis. This conflicting development is related to tendencies in Jung himself that are evident in his own life and in research conducted into the writing and publication of Memories, Dreams, Reflections. Though the status of the latter as Jung's autobiography has been called into question there remains the necessity to explain the myth of Jung's life enshrined there and the impact this has had on a public looking for meaning in a time of considerable change. PMID- 14533395 TI - Jung's mediatory science as a psychology beyond objectivism. AB - In this paper, the author argues that Jung's non-objectivist--yet scientific- epistemology and his empirical/hermeneatic methods of inquiry situate him within a psychological tradition that, in many respects, began with William James and, today, is finding expression in the work of many non-Jungian cognitive scientists. In an effort to encourage dialogue between Jungians and scholars within related intellectual traditions, the author presents evidence from the corpus of Jung's work that demonstrates that, like William James, Jung intentionally rejected the absolutist claims of objectivism and the opposite position on 'anything goes' relativism, emotivism, or subjectivism. Instead, Jung forged a path that led to the meta-psychological position similar to internal realism (Putnam 1981) or experientialism (Lakoff 1987) and to a theoretical psychology that gave a central place both to unconscious cognitive structure and to imagination. This he labelled a 'mediatory science'. The psychological theories developed within this mediatory science framework represent an early articulation of key constructs that are currently used by a number of cognitive scientists seeking to understand how we make sense of experience. PMID- 14533396 TI - The Orpheus complex. AB - This paper examines the possible psychological implications of two adaptations of the myth of Orpheus and Eurydice, both of which were completed in 1997. The first is by a man: 'Deconstructing Harry', a film by Woody Allen. The second is by a woman: 'Eurydice in the Underworld', a short story written by Kathy Acker in the last year of her life. The paper argues that there are only four 'necessary events' in the myth of Orpheus and Eurydice. It defines the sequence of these events as a 'mythic pattern' that represents the experience of loss, unconscious yearning, depression, and psychological inflation. The film is examined as an expression of an 'Orpheus complex', the short story as an expression of an 'Eurydice complex'. The paper suggests a possible reason for the persistence of interest in the myth throughout the twentieth century. Although it notes that women appear to find it easier to free themselves from identification with the mythic pattern, it also provides reasons for thinking that men may be about to do the same. PMID- 14533397 TI - Active imagination and the analysis of film. AB - The emotions aroused by the pleasure-generating mechanisms of the cinema are usually benign because they refer to a virtual world. Experiencing affects aroused by fictions can resemble being drawn into a rehearsal for a possible, imagined future that just might (but more likely never will) occur in the individual's life in the real world. Movie-goers, no less than the readers of novels, actively engage with the symbols that film makers inscribe in the screen text. In so doing, they take a measure of control over elements of the fiction. Screened fiction has thus the potential to help the individual grow in self awareness. Shifting its focus to the work of the film critic, the paper argues that, suitably adapted to the requirements of working with a pre-recorded film or tape, Jung's concept of the active imagination helps us to model, license, limit and endorse the subjective element in textual analysis. Because the method proposed makes it possible formally to recognize that the interpretation of fiction is inevitably integrated with the analyst's personality, interpretation is always a matter of a reader bringing a text into being rather than disinterring a pre-existing object. PMID- 14533398 TI - The prospects for a Jungian psychology in China. AB - The possibility of a Jungian psychology developing in China is considered by a brief historical excursion through the early translations of psychoanalytical works. Translation problems and the contentious nature of some of Freud's ideas have made for their difficult reception in China. The inattention to Jung's ideas in universities in the west in the past, and a reliance on science based subjects by Chinese students studying abroad, have meant little opportunity to study Jung, and, by implication, to translate him. The turbulent political climate in China over the course of the past century has also hindered developments in psychology generally. In addition, certain traditional practices of understanding mind-body relationships and reporting 'illnesses' have precluded the possibility of any psychotherapeutic psychology emerging. However, the present climate looks more favourable for the dissemination of Jungian knowledge, but the question of an appropriate context and a receptive readership remains. Certain Jungian notions can be seen to fit comfortably within traditional systems of Chinese thought but the present day psychology department in China is no more a congenial environment for Jungian psychology than its counterpart in the west. It may be that the success of importing Jungian ideas into China rests with those with a predilection towards arts and cultural sciences, and with the innovations of the organizers of conferences. PMID- 14533399 TI - Dismemberment and reintegration: Aztec themes in contemporary Mexican practice. AB - In a depth process the unconscious presents images of destruction. In my analytic work with a female Mexican artist, the themes of destruction and creation alive in her psyche echo the motifs of the pre-Hispanic myth of Coyolxauhqui. I will illustrate the mythological amplifications of the patient's personal material with illustrations of her paintings. I will also discuss how this myth operates in a male patient, and between the masculine and feminine in Mexican culture. Similar images by the German artist Anselm Kiefer refer to the dismembering of the newly discovered lands and government violence against the youth of Mexico. PMID- 14533400 TI - Pre-Hispanic perspectives on the modern Mexican psyche; contemporary subjects and ancient objects. A Mayan text of evolution: the stages of creation in the Popol Vuh or sacred book of the Mayans. AB - The ancestral tribes of Mexico, like any people living within a certain culture, may be affected by the archetypal images and values of their surroundings. Access to the imagery of the Mayan creation myth, the Popol Vuh, has provided an orientation in my analytical work with Mexican patients as they attempt to recreate themselves by engaging their conflicts around their reality, individuality and capacity to relate. I will address the psychological meaning of the different stages of creation: the original creative event, the man of mud, the man of wood, the false sun and the man of corn. With these images, I will illustrate their clinical application in analytic work with three Mexican middle age male patients dealing with different issues of identity and with a young female patient struggling to separate from a manipulative and destructive family system. PMID- 14533401 TI - 'Is analytical psychology a religion?'. PMID- 14533402 TI - Transplantation ethics: old questions, new answers? PMID- 14533403 TI - Putting patients first in organ allocation: an ethical analysis of the U.S. debate. PMID- 14533404 TI - Public policy, public opinion, and consent for organ donation. PMID- 14533405 TI - The moral status of preferences for directed donation: who should decide who gets transplantable organs? PMID- 14533406 TI - Autonomy's limits: living donation and health-related harm. PMID- 14533407 TI - Procreation for donation: the moral and political permissability of "having a child to save a child". PMID- 14533408 TI - Respecting bodies and saving lives: Jewish perspectives on organ donation and transplantation. PMID- 14533409 TI - Transplantation ethics: old questions, new answers? CQ sources/bibliography. PMID- 14533410 TI - Response to "abortion and assent" by Rosamond Rhodes (CQ Vol 8, No 4) and "abortion, disability, assent, and consent" by Matti Hayry (CQ Vol 10, No 1). Assent and selective abortion: a response to Rhodes and Hayry. PMID- 14533411 TI - Before pigs' germs fly: xenotransplantation and a call for federal action. PMID- 14533412 TI - The new Dutch law on legalizing physician-assisted death. PMID- 14533413 TI - "Help me die". PMID- 14533414 TI - Coming of age! PMID- 14533415 TI - Standardized screening of elderly patients' needs for social work assessment in primary care: use of the SF-36. AB - Fewer hospitalizations and decreased lengths of stay in the hospital have resulted in an increased need for extensive support services and continuing care planning for elderly people in primary care. Early identification of elderly patients needing community and hospital nonmedical services is necessary so that timely appropriate services can be delivered. This study addresses the issue of whether a standardized health-related quality of life questionnaire, the SF-36, can be used independently as a screen predicting primary care elderly patients' needs for social work assessment. In addition, the question of what scales on the SF-36 a social worker would use to screen patients in need of assessment is explored. PMID- 14533416 TI - Treatment of forensic patients: an expanding role for public psychiatric hospitals. AB - This article explores the increased role of state-operated public psychiatric hospitals in treating forensic patients. Patients with a forensic legal status have a mental illness and are involved with the criminal justice system. Using data from Missouri's forensic system, the article compares the characteristics of voluntary hospital patients with those of the largest hospitalized group of forensic patients--those found not guilty by reason of insanity (NGRI). Overall, NGRI patients tended to be higher functioning, less likely to have committed assaultive acts, and more likely to have substance abuse and personality disorder diagnoses. The article explores the treatment needs of forensic patients who reside in public psychiatric hospitals and discusses the effect of the strong presence of forensic patients in public psychiatric hospitals on social work practice, including clinical work with forensic patients, social work administration, and social work advocacy. PMID- 14533417 TI - Through indigenous eyes: Native Americans and the HIV epidemic. AB - This article examines the phenomenon of HIV within the context of a Native American culture. Native Americans have some risk factors for HIV transmission that differ from those found in other populations. In addition, prevention and intervention activities with this population must consider cultural variables to maximize their effectiveness. Brief anecdotes are used to illustrate various concepts related to HIV and Native Americans and to include a human face along with facts and statistics. The author's unique perspective, coupled with a broad discussion of relevant issues enables non-Native American readers to understand better the phenomenon of HIV as it exists within a Native American context. PMID- 14533418 TI - Promises in the plague: passage of the Ryan White Comprehensive AIDS Resources Emergency Act as a case study for legislative action. AB - In a conservative political climate, it is very important that social workers and others who are concerned with social welfare understand how to affect social change through legislative advocacy. This article presents a case study in successful intervention using the passage by the Congress of the Ryan White Comprehensive AIDS Resources Emergency (CARE) Act of 1990. The major influences in the passage of this landmark bill were building a diverse coalition that provided a very broad base of advocacy; securing bipartisan support; using recognizable personalities to call attention to the problem; and defining the issue as one that affected people from all socioeconomic and ethnic groups. PMID- 14533419 TI - The social worker in the emerging field of home care: Professional activities and ethical concerns. AB - Although home care is one of the fastest growing segments of the health care delivery system, there is little empirical data on the professional functions or ethical concerns of social workers employed in home care. To provide this information 118 social workers in proprietary and nonprofit agencies in one midwestern and one southern state were surveyed. The results indicated that social workers performed a wider array of professional functions with a more diverse population of patients than had been documented previously. Workers experienced ethical concerns related to self-determination, barriers to access of services, implementing advance directives, and assessment of mental competence most often. Workers in proprietary agencies rated the ethical concern of barriers to access of services as occurring significantly more often than workers in nonprofit agencies. PMID- 14533420 TI - Preparation for oncology settings: what hospice social workers say they need. AB - According to recent data, 78 percent of hospice social workers' direct practice hours are spent with patients and families diagnosed with terminal cancer. A national sample of 108 hospice social workers participated in an exploratory study investigating the influence of their formal academic training, informal on the-job training, and related professional and personal experiences on their preparation for oncology social work. Findings suggest that social workers view both the generalist or core courses as well as the specialist and elective courses as important in preparing them for oncology social work. In addition, on the-job training focusing on medical terminology, interdisciplinary, collaboration, death and dying and the integration of personal experiences are important for continuing growth and success in this work. Implications for social work practice, research, and formal and informal educational opportunities are discussed. PMID- 14533421 TI - Social work in psychiatric home care: regulations, roles, and realities. AB - Home care services for people with psychiatric diagnoses are increasing in popularity with consumers, providers, and insurance companies. Traditionally, home care social workers have not provided in-home clinical services because of restrictive regulations and industry culture. This article presents strategies for establishing a clinical role for social workers in a psychiatric home care program that meets Medicare, Joint Commission on Accreditation of Healthcare Organizations, and NASW standards. A discussion of skills requirements, appropriate case assignment, professional role clarifications and differentiations, and marketing strategies is presented. PMID- 14533422 TI - The aging are coming and they are us. PMID- 14533423 TI - Spirituality and political correctness. PMID- 14533424 TI - Are nurses ready to take over junior doctors' roles? PMID- 14533425 TI - The higher education debate--reignited. PMID- 14533426 TI - Students' childcare needs should be provided for. PMID- 14533427 TI - Should nursing students be paid a salary? PMID- 14533428 TI - Student life. The real picture. PMID- 14533429 TI - An overview of permanent and temporary cardiac pacemakers. AB - As pacemaker technology comes closer to mimicking the heart's own pacing system, patients in need of such devices can hope to live increasingly normal and productive lives. This article outlines common types of cardiac pacing and indications for pacemaker insertion. Nursing considerations are outlined both during and following the insertion of a pacemaker. PMID- 14533430 TI - What you need to know about ... glomerulonephritis. PMID- 14533431 TI - Subcutaneous injections. PMID- 14533432 TI - Patient use of the Internet to obtain health information. AB - With increasing use of the internet by patients, questions are being raised in the press and by professionals on whether members of the public may be harming themselves by gaining access to previously hidden material. A search of the literature from a variety of sources revealed that the evidence was conflicting. Because the internet is here to stay, it is suggested that health professionals use it in partnership with patients, at the same time acknowledging its pitfalls. PMID- 14533433 TI - Health care assistants: developing their role to include nursing tasks. AB - Detailed information about the educational needs and aspirations of health care assistants in acute care settings is scarce. A study was undertaken across four acute trusts to develop an advanced health care assistant training programme. The findings support the drive to further develop the role of this workforce group. PMID- 14533434 TI - A guide to surviving your student years. PMID- 14533435 TI - Patients are paying the price. PMID- 14533436 TI - How to ... write a great essay. PMID- 14533437 TI - Biweekly low-dose sequential gemcitabine, 5-fluorouracil, leucovorin, and cisplatin (GFP): a highly active novel therapy for metastatic adenocarcinoma of the exocrine pancreas. AB - Phase II studies have suggested an improved response rate and acceptable toxicity profile associated with gemcitabine combinations compared to gemcitabine alone for treatment of metastatic adenocarcinoma of the pancreas. The GFP regimen (gemcitabine, 5-fluorouracil, leucovorin, and cisplatin) is based on laboratory evidence of disease-specific chemotherapy interaction. This retrospective analysis examined the outcome of 49 consecutive patients with histologically confirmed metastatic pancreatic adenocarcinoma treated between July 1998 and September 2000. Day 1 treatment consisted of gemcitabine 500 mg/m2 over 30 minutes and then leucovorin 300 mg bolus, 5-fluorouracil (5-FU) 400 mg/m2 bolus, followed by infusional 5-FU 600 mg/m2 over 8 hours. Day 2 consisted of leucovorin 300 mg bolus, 5-FU 400 mg/m2 bolus, followed by cisplatin 50-75 mg/m2 over 30 minutes and then infusional 5-FU 600 mg/m2 over 8 hours. Treatment was administered every 2 weeks. Median patient age was 61.5 years, 74% were men, and 20 patients had refractory disease (11 patients had disease progression upon gemcitabine-based therapy). Grade 3-4 toxic effects (% patients) consisted of neutropenia (30%), thrombocytopenia (14%), anemia (8%), and neutropenic fever (2%). Grade 3-4 nonhematological toxicities (% patients) consisted of neuropathy (14%), ototoxicity (8%), nephrotoxicity (6%), nausea/vomiting (14%), and mucositis (10%). The majority of dose reductions were made for neuropathy or cytopenias. Filgrastim and erythropoietin were given as needed to promote dose intensity. Eight patients attained a partial response (PR) by RECIST criteria. Fourteen had stable disease (SD). Two patients attaining PR and two attaining SD had progressive disease with prior gemcitabine-based therapy. The median time to disease progression (TTP) from GFP start was 9 weeks. For all 49 patients, the median overall survival (OS) from GFP start was 10.6 months, 12-month survival was 46%, and 24-month survival was 30%. Notably, upon disease progression, 31 patients continued to receive the GFP regimen with irinotecan 80 mg/m2 inserted on day 1 following gemcitabine, the G-FLIP regimen (gemcitabine, 5-fluorouracil, leucovorin, irinotecan, and cisplatin). Measured from G-FLIP initiation, the TTP for the 31 patients treated sequentially was 10 weeks, and for the 14 patients attaining SD or PR the TTP was 25 weeks. The median overall survival measured from GFP initiation was 11.8 months. The response rate, non-cross resistance, TTP, OS, and tolerability warrant prospective development of this novel combination. This experience also demonstrates that adding a single new drug such as irinotecan to the same first-line chemotherapy combination upon disease progression may be an important alternative for the treatment of relapsed/resistant cancer. PMID- 14533438 TI - Vinorelbine and cisplatin for metastatic breast cancer: a salvage regimen in patients progressing after docetaxel and anthracycline treatment. AB - PURPOSE: To assess the antitumor efficacy and safety of a combination of vinorelbine (VNR) and cisplatin in patients with metastatic breast cancer previously treated with anthracyclines and docetaxel. PATIENTS AND METHODS: Thirty-six patients with assessable metastatic breast cancer previously treated with anthracyclines and docetaxel (adjuvant n = 1, palliative n = 20, both n = 15) were studied. Cisplatin was given at 75 mg/m2 on day 1 followed by 25 mg/m2 VNR on days 1 + 8 in a 5-minute i.v. infusion. Courses were repeated every 3 weeks. Treatment was continued until disease progression, excess toxicity, or patient refusal. Patients were classified according to their response to anthracyclines according to criteria published previously: 1) Anthracycline and/or docetaxel resistant were patients who progressed during treatment with anthracyclines and docetaxel or within 4 months after cessation of treatment (metastatic). In addition, adjuvant patients who progressed within 6 months after completion of chemotherapy belong to this group. 2) Anthracycline and/or docetaxel relapsed were either metastatic patients who responded initially and then progressed after 4 months of completing an anthracycline- and docetaxel based chemotherapy or patients who progressed after 6 months from completion of anthracycline/docetaxel-based adjuvant chemotherapy. RESULTS: Two patients (5.6%) achieved a complete response (CR) and 15 patients (41.6%) achieved a partial response (PR), for an overall response rate (OR) of 47.2% (95% confidence interval, 31-63). Of 18 patients relapsed to anthracycline/docetaxel, 2 had a CR (11%) and 8 a PR (44.4%), giving an objective response of 55.5%. Stable disease (SD) was observed in one patient (5.5%); seven patients had progressive disease (PD) (39%). Among the 18 resistant patients, 7 PRs (39%) were observed (p = 0.5), one patient (5.5%) had stable disease, 10 patients (55.5%) progressed. The median time to progression (TTP) was 16 weeks and median overall survival 36 weeks. Relapsed patients had a longer TTP than resistant patients (24 vs. 8 weeks, p = 0.05) but similar survival (48 vs. 24 weeks, p = 0.173). All patients were assessed for toxicity. The main toxicity was neutropenia grade 3 and 4 in 47% of patients. Febrile neutropenia requiring hospitalization was absent. There were no treatment-related deaths. Thrombocytopenia grade 3 and 4 occurred in four patients (11%). Phlebitis, orthostatic hypotension, and asthenia, all reversible, were observed in 3% of patients, respectively. CONCLUSION: This cisplatin/VNR regimen is well tolerated and active in patients who failed anthracyclines and docetaxel treatment. The response rate, TTP, and survival data are high and indicate that cisplatin/VNR may have a place as salvage treatment in this group of patients. If these results can be verified in multi-institutional trials, this combination of drugs would merit investigation as part of a first-line therapy in breast cancer. PMID- 14533439 TI - A phase II study of modified deGramont 5-fluorouracil, leucovorin, and oxaliplatin in previously treated patients with metastatic colorectal cancer. AB - PURPOSE: To evaluate the toxicity and efficacy of a modified deGramont regimen of 5-fluorouracil (5-FU), leucovorin, and oxaliplatin in patients with advanced colorectal cancer who have progressed on at least one but not more than two prior chemotherapy regimens. PATIENTS AND METHODS: Patients with stage 4 colorectal cancer were treated with oxaliplatin 85 mg/m2 by a 2-hour intravenous infusion, followed by leucovorin 500 mg/m2 by a 2-hour intravenous infusion, followed by 5 FU 400 mg/m2 by bolus injection, followed by 5-FU 2.4 g/m2 administered by a 46 hour continuous infusion. Cycles were administered every 2 weeks. RESULTS: Seventy patients were treated and 68 patients had previously received irinotecan. Eleven percent of patients had a partial response, 33% of CEA-evaluable patients had a > or = 50% drop in their CEA level. The median time to progression was 6.2 months, and the median overall survival was 8.7 months. Toxicity was mild to moderate, as 14% of patients experienced grade 3 or 4 neutropenia and 3% of patients experienced grade 3 neuropathy. CONCLUSION: The modified deGramont regimen of 5-FU, leucovorin, and oxaliplatin is tolerable and is associated with a modest degree of antitumor activity in patients who have progressed on both 5 FU and irinotecan. PMID- 14533440 TI - Phase II trial of 96-hour paclitaxel in previously treated patients with advanced esophageal cancer. AB - BACKGROUND: A 96-hour infusion schedule of paclitaxel demonstrates tolerability and antitumor activity in lung cancer and breast cancer refractory to short duration infusion paclitaxel. Given the activity of paclitaxel in esophageal cancer, a phase II trial of 96-hour infusion paclitaxel in esophageal cancer was undertaken. METHODS: Both adenocarcinoma and squamous cell histology were included. Paclitaxel was administered at 140 mg/m2 over 96 hours every 21 days. Patients who had metastatic disease to the liver and transaminases greater than two times normal value received 120 mg/m2. Response to treatment was evaluated after the first two cycles and subsequently every third cycle. RESULTS: Ten men and four women were entered. All were eligible for response and had stage IV disease. Thirteen patients were previously treated. All 13 received prior short duration paclitaxel-containing chemotherapy regimens. Eleven patients had adenocarcinoma and three squamous cell cancer. Patients completed a mean of two cycles (range one to eight) prior to disease progression. No major responses were observed. Toxicity was minimal and included grade 3/4 neutropenia in 14% of patients. One patient with adenocarcinoma demonstrated stable disease for 28 weeks. CONCLUSION: No major activity was observed in a population of previously treated patients. Ninety-six-hour paclitaxel in metastatic esophageal cancer is generally well tolerated with minimal toxicity; however, it is ineffective in previously treated patients. Further evaluation of this schedule of paclitaxel in combination with concurrent radiotherapy, where its radiosensitizing potential may be useful, is ongoing in locally advanced esophageal cancer. PMID- 14533441 TI - Combined therapy with topotecan and gemcitabine in patients with inoperable or metastatic non-small cell lung cancer. AB - PURPOSE: We conducted a phase I/II trial of topotecan combined with gemcitabine in patients with metastatic or unresectable non-small cell lung cancer (NSCLC) based on preclinical data showing in vitro synergy against an established lung adenocarcinoma cell line. The aim was to determine the maximally tolerated dose (MTD) of topotecan when the gemcitabine dose is held constant, as well the dose limiting toxicity (DLTs) of this combination in NSCLC patients. PATIENTS AND METHODS: Twenty-four patients with stage IIIB or IV NSCLC were treated weekly times 3 with a week break with gemcitabine (1250 mg/m2 over 30 minutes) and topotecan (30 minutes) at varying doses. The starting dose of topotecan was 1.0 mg/m2 and doses were escalated in 0.25-mg/m2 increments until the MTD was achieved. RESULTS: The MTD of gemcitabine/topotecan was 1250 mg/m2 of gemcitabine and 2.00 mg/m2 of topotecan (level 5). Neutropenia was the DLT. Few nonhematologic toxicities were observed. There were 5 (21%) partial responses among 24 patients. The median survival was 22 weeks. Two patients have had prolonged (> 2 year) survival. CONCLUSION: The combination of gemcitabine and topotecan seems to be active against NSCLC with acceptable hematologic toxicity and minimal nonhematologic toxicity. The recommended dose for further study is 1250 mg/m2 of gemcitabine (days 1, 8, 15) and 2.0 mg/m2 of topotecan (days 1, 8, 15) administered every 28 days. PMID- 14533442 TI - What are the most important symptom targets when treating advanced cancer? A survey of providers in the National Comprehensive Cancer Network (NCCN). AB - We derived a set of brief, clinically relevant symptom indices for assessing symptomatic response to chemotherapy for advanced bladder, brain, breast, colorectal, head and neck, hepatobiliary/pancreas, lung, ovarian, and prostate cancers. Questions were extracted from a multidimensional cancer quality of life (QOL) measurement system, the Functional Assessment of Cancer Therapy (FACT). Surveys of disease-related symptoms were presented to expert physicians and nurses at 17 National Comprehensive Cancer Network (NCCN) member institutions. In a two-step procedure, each expert narrowed the list to no more than five of the very most important to attend to when assessing the value of drug treatment for advanced disease. Symptoms endorsed at a frequency greater than chance probability were retained for the nine symptom indices. The resulting NCCN/FACT symptom indices are comprised of 6-15 items, depending on disease. Fatigue, pain, nausea, weight loss, worry about worsening condition, and contentment with current QOL were consistently selected by experts as priority symptoms across tumor sites. These nine tumor-specific symptom indices indicate the most important clinician-rated targets of chemotherapy for many advanced cancers. These results await validation in patient populations and examination of the extent to which changes in symptomatology translate into meaningful improvement to the patient. PMID- 14533443 TI - Detection of recurrent or residual nasopharyngeal carcinomas after radiotherapy with technetium-99m tetrofosmin single photon emission computed tomography and comparison with computed tomography--a preliminary study. AB - The effectiveness of technetium-99m tetrofosmin (Tc-TF) single photon emission computed tomography (SPECT) of the head and neck for detecting recurrence of nasopharyngeal carcinomas (NPC) was evaluated and compared with computed tomography (CT). Both Tc-TF SPECT and CT of the head and neck were performed in 36 NPC patients 4 months after radiotherapy. All 36 then underwent histopathological examinations of nasopharyngeal biopsies. Based on the histopathological results, the sensitivity, specificity, and accuracy of visually interpreted Tc-TF SPECT images, to differentiation of recurrent NPC from benign lesions, were 64%, 96%, and 86%, respectively. The sensitivity, specificity, and accuracy of CT for detecting recurrent NPC were 73%, 88%, and 83%, respectively. The Tc-TF SPECT has a better specificity but a lower sensitivity to differentiate benign lesions and recurrent/residual NPC when compared with CT. Combined Tc-TF SPECT and CT sensitivity and specificity were 100% and 88%, respectively. There are much better either test alone. However, further studies, including a larger NPC patient population, are warranted to determine the exact role and clinical usefulness of Tc-TF SPECT to differentiate benign lesions and recurrent/residual NPC. PMID- 14533444 TI - A phase I human trial of mitoguazone and gemcitabine sequential bi-weekly treatment of cancer patients. AB - Our previous studies have demonstrated the existence of synergism in a combination therapy using mitoguazone and gemcitabine when the mitoguazone is administered 24 hours before gemcitabine. Based on the cell culture and animal experimental results, a phase I clinical trial was performed in order to determine the toxicity of the combined treatment. Mitoguazone and gemcitabine were administered sequentially: mitoguazone on day 1 and gemcitabine on day 2. This cycle was repeated every 2 weeks. The dosages of these two drugs were varied between patients. Ten patients were enrolled in the study. Six patients began treatment at dose level 1 (mitoguazone 500 mg/m2, gemcitabine 1500 mg/m2), three at dose level 2 (mitoguazone 500 mg/m2, gemcitabine 2000 mg/m2), and one at dose level 3 (mitoguazone 600 mg/m2, gemcitabine 2000 mg/m2). Dose-limiting toxicity (DLT) was only observed in two patients treated at dose level 1 and one patient treated at dose level 3, while all the other patients only experienced nonhematologic toxicity, such as asthenia and mucositis. Two melanoma patients showed responses (one partial and one minor) to the treatment. One lymphoma patient also showed a brief partial response. This phase I trial indicated that the combination of mitoguazone and gemcitabine had limited but noticeable activity for treatment of cancer patients. Further study on the toxicity and on the effect of the scheduled mitoguazone-gemcitabine combination is needed. PMID- 14533445 TI - Epiphora induced by intermittent docetaxel (taxotere) in patients with non-small cell lung cancer. PMID- 14533446 TI - Gastrointestinal stromal tumors: a paradigm for molecularly targeted therapy. PMID- 14533447 TI - Symptom control. AB - Symptom control has become increasingly recognized as an important goal in patient care. In this article, advances in symptom assessment, and various definitions of symptom improvement are reviewed. Theoretical concepts underlying symptom control and clinically significant change are presented, as well as the role of symptom control as an endpoint in clinical trials. Symptom control is then surveyed in two broad categories for selected symptoms. The first area is therapy related symptoms, secondary to chemotherapy, radiation, hormonal therapy, and surgery. Symptoms reviewed include chemotherapy related mucositis, emesis, fatigue; hot flashes; and radiation related dermatitis, xerostomia, and mucositis. The second area is palliative oncologic approaches to disease-related symptoms. Results in palliative chemotherapy, palliative radiation therapy, cancer pain, and lack of appetite are summarized. Areas requiring further research are noted. Findings are presented in both a clinical and research context to help guide the reader with interpreting symptom control studies. PMID- 14533448 TI - Leukemia in severe congenital neutropenia: defective proteolysis suggests new pathways to malignancy and opportunities for therapy. AB - Severe congenital neutropenia (SCN), a heterogeneous disorder that includes Kostmann syndrome, predisposes to myelodysplasia and acute myelogenous leukemia. Recently identified heterozygous mutations in the gene ELA2, encoding neutrophil elastase on human chromosome 19pter, account for the majority of autosomal dominant cases of SCN, including those demonstrating neoplastic progression. The involvement of the serine protease neutrophil elastase, localized to the granules of neutrophils and monocytes, implies an unexpected role for proteolytic regulation of hematopoiesis. Continued elucidation of the clinical features, molecular genetics, and biochemistry is likely to provide insight into novel pathways of leukemia induction with attendant prospects for new avenues of therapy. PMID- 14533449 TI - Psychosocial issues associated with genetic testing for breast and ovarian cancer risk: an integrative review. AB - The identification of the BRCA1/2 genes, and their possible etiologic relationship with various forms of inherited cancer, has been recognized universally as a cornerstone in the search for cancer's genetic link and has made it possible to identify specific individuals and families who harbor a mutation in one of these predisposition genes. Genetic testing for breast and ovarian cancer susceptibility may pose unanticipated psychological and social problems. Because of the recent availability of predisposition genetic testing, research efforts have begun to investigate factors that may influence an individual's intention to undergo testing and the psychosocial sequelae associated with testing. The purpose of this article is to provide an integrative review of the literature that will delineate what is currently known about the psychosocial issues associated with genetic testing for breast and ovarian cancer risk. Important generalizations from the literature include: (a) a positive test for breast cancer susceptibility may ignite a psychological response similar to the diagnosis of breast cancer itself; (b) there is likely a subset of individuals at increased risk for hereditary breast and ovarian cancer who are also at risk for sustained psychosocial problems; (c) available literature challenges a common notion that only individuals with a positive test result will need psychosocial services; and (d) at-risk individuals are basing health care decisions on genetic testing information, thus they are making important decisions under conditions of uncertainty. Clinical issues and directions for future research were highlighted. PMID- 14533450 TI - UFT and oral leucovorin as radiation sensitizers in rectal and other gastrointestinal malignancies. AB - UFT is the combination of the fluorouracil (5-FU) prodrug tegafur, and the dihydrophyrimidine dehydrogenase inhibitor uracil. UFT and leucovorin have been used in the oral treatment of a variety of malignancies. Given the radiosensitizing properties of UFT, its combination with radiation therapy is a logical step in the development of novel strategies for the treatment of gastrointestinal malignancies. Several phase I and phase II studies, described in this review, have attempted to substitute UFT for 5-FU in chemoradiation strategies for the treatment of patients with esophageal, pancreatic and rectal cancer. The results thus far suggest that this approach has acceptable toxicity and promising efficacy. Given the availability of the novel oral fluorinated pyrimidines and their efficacy in metastatic diseases, it remains to be determined whether 5-FU will eventually be substituted by oral fluorinated pyrimidines in the adjuvant and chemoradiation settings. PMID- 14533451 TI - Cancer pharmacogenomics: SNPs, chips, and the individual patient. AB - There is great heterogeneity in a patient's response to medications, often requiring empirical strategies to define the appropriate drug therapy for each patient. Pharmacogenomics aims to elucidate further the inherited nature of interindividual differences in drug disposition and effects, with the ultimate goal of providing a stronger scientific basis for selecting the optimal drug therapy and dosage for each patient. These genetic insights should also lead to mechanism-based approaches to the discovery and development of new medications. Genetic polymorphisms in drug metabolizing enzymes, transporters, receptors, and other drug targets have been linked to interindividual differences in the efficacy and toxicity of many medications. For example, polymorphism in thiopurine methyltransferase (TPMT) results in altered degradation of the commonly prescribed agent 6-mercaptopurine. This genetic variant has significant clinical implications because patients with functionally relevant homozygous mutations in the TPMT gene experience extreme or fatal toxicity after administration of normal doses of 6-MP. In addition, patients heterozygous for mutations in TPMT require slight dosage reduction of 6-MP and experience a greater degree of systemic toxicity from the agent. This and other examples of genetic polymorphism relevant to the treatment of cancer are highlighted to illustrate the promise and pitfalls of the exciting area of cancer therapeutics, with the potential of providing a stronger scientific basis to optimize drug therapy on the basis of each patient's genetic constitution. PMID- 14533452 TI - Novel perspective: focusing on the X chromosome in reproductive cancers. AB - In an XX female, one of the two X chromosomes has been inactivated during early embryonic life to achieve a compensation of X-linked gene products between males and females, leaving only one allele of X-linked genes functional. There are some X-linked genes escaping the X-inactivation, i.e., being expressed from both alleles. Escape from X-inactivation varies at different levels; some genes have both alleles active in some women but only one allele active in others, whereas some other genes have both alleles active in neoplastic tissue but only one allele active normally. The X-inactivation may be considered functionally equivalent to a loss of heterozygosity (LOH) for some genes, whereas escape from X-inactivation may be equivalent to functional gene amplification for others. The physiological LOH may make X-linked tumor suppressor genes lose their function more easily, compared with autosomal tumor suppressor genes, thus predisposing women to cancer formation more easily. Moreover, the human X chromosome contains many genes related to cancer or to sex and reproduction. All these properties of the X chromosome suggest that it may play more important roles than any autosomal chromosome in the development and progression of reproductive and urologic cancers. PMID- 14533453 TI - 96-hour paclitaxel infusions: at least 93 hours too long. PMID- 14533454 TI - Has the moment arrived for symptoms? PMID- 14533455 TI - Breast cancer in HIV positive women: a report of four cases and review of the literature. PMID- 14533456 TI - 2004 JCAHO National Patient Safety Goals approved. PMID- 14533457 TI - Revisions to assessment and environment of care requirements for assisted living. PMID- 14533458 TI - Subacute care requirements integrated into general long-term care standards. PMID- 14533459 TI - Preventing surgical fires. PMID- 14533460 TI - [Dietary calcium and the prevalence of spinal deformities in a sample of the elderly population]. AB - The aim of the study was to determine the role of calcium intake through nutritional consumption of milk and other milk products currently and in three time periods during life (until 25 years, 25-50 years and 50 years and over) and to relate the results to finding of vertebral deformities in an unselected sample of population of Zagreb (Croatia), aged 50 years and above. Each of 425 recruited individuals (269 women and 156 men) completed interviewer administered questionnaire and had lateral thoracolumbar radiography performed according to a standard protocol. The questions related to number of days in which each subject consumed milk and its products in the last week, and milk consumption in three life-periods. The morphometric measurements of vertebral height were used to determine the occurrence of vertebral deformity according to the method described by McCloskey et al. Adding numbers of days in which each subjects in the last week consumed milk and its products the score ranging from 0-28 was obtained. Between two groups of subjects (group I = score 0-10 and group II = score 11 and over), there was no difference by age, but there was by sex, women being those with greater current consumption of milk products. There was an increased risk of finding vertebral deformity in women appertaining to group I (OR = 3.10, 95% CI = 1.20-7.97), but not for men (OR = 0.93, 95% CI = 0.37-2.35), nor for both sexes (OR = 1.89, 95% CI = 0.99-3.63). There was a statistically significant difference in frequency of drinking milk in period 25-50 years of life, women being again more frequent consumers, but not in other periods of life. There was no correlation in frequency of milk consumption and prevalence of vertebral deformity in any of the periods of life, separately for men and women and taken together. To estimate milk consumption throughout the whole life we added the figures for all three periods of life, getting the score ranging from 3-12. The subjects were divided into two groups, and no increased risk of vertebral deformity was found in those who had been drinking less milk (OR = 1.43, 95% CI = 0.77-2.66). In our study women with currently less dietary calcium intake have greater risk for vertebral deformities. No association was found between frequency of milk consumption throughout the life and prevalence of vertebral deformities. PMID- 14533461 TI - [Prevalence of gallstones in patients on hemodialysis]. AB - End-stage renal disease affects all organ systems. Nevertheless, frequency of some pathologic conditions does not differ in end-stage renal disease patients in comparison with their age- and sex-matched controls recruited from general population. The paper presents the results of the survey on the prevalence of gallstones in a relatively big group of hemodialysis patients (n = 114; 49 females, 65 males). The goal of the study was to examine whether the genesis of gallstones could be explained by renal failure itself. Gallstones have been found in 24 of 114 hemodialysis patients (21.05%), predominantly females (n = 14). Gallstones have been found in 28.75% of female patients, and in only 15.38% of male patients. The frequency of gallstones correlated significantly with age of the patients (r = 0.26, p < 0.05), number of pregnancies (r = 0.20, p < 0.005) and deliveries (r = 0.22, p < 0.05). Patients suffering from gallstones were significantly older (60.58 +/- 11.99:51.72 +/- 14.01 years, t = 3.10, p < 0.01), with every third older than 54 affected. The results observed do not differ from the results of other authors that report the morbidity in general population. In conclusion, gallstones in hemodialysis patients show same frequency as in general population and probably result from the same pathophysiologic mechanisms. PMID- 14533462 TI - [Isolated cervical spinal cord hemangioblastoma--case report]. AB - Hemangioblastomas are rare tumors which account for 0.9-2.1% of central nervous system neoplasms. The most common site of hemangioblastomas is the cerebellum, while they are rarely located in spinal cord, cerebrum and brain stem. Hemangioblastomas occur as a sporadic entity, and as a manifestation of von Hippel-Lindau syndrome. A 33-year old patient with isolated cervical spinal cord hemangioblastoma is presented. PMID- 14533464 TI - [Streptococcus suis infection, a zoonosis we should have in mind--2 case reports]. AB - Streptococcus suis is primarily pig's pathogen, but can cause disease in a man exposed to contact with pigs (butchers, abatoir workers, farmers). Infection in man is most frequently manifested as purulent meningitis, with deafness and ataxia, but there are rare reports on septic shock with multiple organ failure and death. We report two patients with Streptococcus suis type 1 infection, treated in "Dr Josip Bencevic" General Hospital, Slavonski Brod. The first patient suffered a very abrupt and severe illness, with septic shock, multiple organ failure and lethal outcome. The second patient had purulent meningitis with deafness. Both of them had a defect in immunologic function before infection. They were probably infected during manipulation with pork meat at home. Our patients had infection with Streptococcus suis type 1, contrary to reports on prevalent human infection with type 2. It is necessary to make epidemiologic survey on human infection with that pathogen, especially in those professionally exposed to pigs and pork meat. PMID- 14533463 TI - [Bronchiolitis obliterans with organizing pneumonia in a patient treated with amiodarone]. AB - Bronchiolitis obliterans organizing pneumonia (BOOP) is a well-defined clinicopathological entity. The aetiology of BOOP is generally unknown, although it has been associated with specific diseases or various pharmaceutical drugs. The amiodarone is one of them. We report a patient with BOOP secondary to amiodarone therapy, who presented with cough, fever and sputum production, dyspnoea and night sweats lasting for two months. A chest radiograph showed bilateral patchy and interstitial infiltrates. Lymphocyte phenotyping of bronchoalveolar lavage fluid showed decreased ratio of CD4+:CD8+ lymphocytes. Transbronchial lung biopsy established the diagnosis of BOOP. After stopping amiodarone therapy, symptoms disappeared and the chest radiograph remained normal within two months. PMID- 14533465 TI - [Loco typico radius fractures--personal experience]. AB - Fractura radii loco typico (FRLT) is the fracture of the distal radius. That is the one of the most frequent fractures of locomotor system with the widest range of treatment in traumatology. Therapy depends on the stability of the fracture: nonoperative or operative. We analysed the five-year experience of our Department of Traumatology where more than 1500 patients with FRLT have been treated in the urgent surgery clinic and 126 of them were hospitalized. In our study we evaluated the results of the operative treatment and postoperative functional status of a treated wrist. According to the A-O classification, we hospitalized 36 patients with A type, 28 patients with B type, and 62 patients with C type of FRLT. We operated 80 patients. The postoperative functional status of a treated wrist was excellent or good in 64 patients and good in 16 patients. The treatment of FRLT depends on the type and the complications of the fracture and the age of the patients. Operative therapy is indicated in the unstable FRLT or after an inadequate nonoperative treatment. PMID- 14533466 TI - [Bronchoalveolar lavage]. AB - Bronchoalveolar lavage (BAL) is a bronchoscopic technique that reveals specific insight in the distal parts of lung parenchyma. During the past twenty years, research of cellular and extracellular bronchoalveolar profiles gave important information on pathogenesis of some pulmonary disorders, promoting this technique as a diagnostic tool in pulmonary infections and interstitial lung diseases. Bronchoalveolar lavage is a safe, well-tolerated and suitable diagnostic procedure in immunocompromised patients. The patients with alveolar proteinosis gain therapeutic benefit of bronchoalveolar lavage. In this article we described technical notes, sampling, storage, cellular and noncellular analyses of bronchoalveolar lavage, including interpretation of results and significance in pulmonary diseases. PMID- 14533467 TI - [Effects of organic aerosols on the respiratory system in occupational exposure]. AB - The effects of organic dusts on the respiratory system of workers employed in food processing, in textile workers, as well as in agricultural and livestock farmers are reported. Deposition and elimination of particles from the lung as well as the possible mechanisms of the organic dust effects on respiratory system are described. Separately are presented results of our investigations on the prevalence of respiratory symptoms and diseases as well as lung function tests as indicators of the condition of the respiratory system. Data on the experimental studies with aqueous extracts of organic dusts on smooth muscle of isolated guinea pig trachea suggest that the bronchoconstrictive effects in humans can be reproduced in nonsensitized guinea pigs. The preventive measures in the development of respiratory impairments in subjects occupationally exposed to organic dusts are described. PMID- 14533468 TI - [Hepatitis B vaccination in patients on chronic hemodialysis]. AB - Patients on chronic haemodialysis (PCHD) respond less well to vaccination with recombinant hepatitis B virus superficial antigen (HbsAg) because of immunity disorders in uraemic patients. Today many schemes and vaccination modification for nonresponding PCHD are proposed. The reaction on vaccination with HbsAg is weaker in those PCHD who had diabetes, older age and insufficient nutritive parameters. In those patients some alternative schemes of vaccinating for nonrespondering PCHD must be considered, especially one of the proposed intradermal ways of vaccine inoculation. PMID- 14533469 TI - [Rapid identification of methicillin-resistant strains of Staphylococcus aureus (MRSA)]. PMID- 14533470 TI - [Importance of hospital registries in cancer research and the pathologist's role]. PMID- 14533471 TI - The Royal College of Nursing Research Society: nurses and research ethics. PMID- 14533472 TI - Research governance and students: what are NHS trusts looking for? AB - Student research in the National Health Service (NHS) must now be conducted within the framework of research governance and research ethics. In this paper, Tony Soteriou and Gill Hek highlight the considerations that undergraduate, post registration and postgraduate students, as well as their supervisors and course lecturers need to make at the outset of planning student research in the NHS. PMID- 14533473 TI - Research governance and postgraduate nurse education: the tensions and some solutions. AB - In this paper Lorraine Ellis and Sue Peckover discuss the implications of the current NHS Research Governance Framework for the supervision of Masters level students undertaking empirical research as part of their programme of study. The paper highlights some of the inherent tensions resulting from the introduction of research governance, and offers some solutions. PMID- 14533474 TI - Research philosophy: towards an understanding. AB - In this paper, Frank Crossan argues that the distinction between quantitative and qualitative philosophies and research methods is sometimes overstated, and that triangulation of methods in contemporary research is common. It is, therefore, important to understand the strengths and weaknesses of each approach, and this paper aims to provide the novice researcher with a basis for developing that understanding. A descriptive analysis of the philosophies of positivism and post positivist thinking in relation to research methodology is presented both as an introduction to the philosophical basis of research, and as a sound basis from which to discuss the 'quantitative-qualitative' debate. PMID- 14533475 TI - Discovering the literature of nursing: a guide for beginners. PMID- 14533478 TI - Antimicrobial activity of some 2- and 3-pyridinyl-1H-benzimidazoles and their FeIII, CuII, ZnII, and AgI complexes. AB - 2-(2-Pyridinyl)- (LI), 2-(6-methyl-2-pyridinyl)- (LII), 2-(6-methyl-2-pyridinyl) 5-methyl-(LIII), 2-(3-pyridinyl)- (LIV), 2-(3-pyridinyl)-5-methyl-1H benzimidazoles (LV) and their complexes with Fe(NO3)3, Cu(NO3)2, Zn(NO3)2, and AgNO3 were synthesized and antibacterial activity of the compounds was tested toward Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella typhi, Shigella flexneri, Proteus mirabilis and antifungal activity against Candida albicans. The methyl groups of LIII increase the antimicrobial activity. The AgI complexes have considerable activity toward the microorganisms. Some ZnII complexes show an antimicrobial effect against S. aureus and S. flexneri, although the ligands themselves have no effect. CuII complexes have a considerable antibacterial effect to S. aureus and S. epidermidis. PMID- 14533477 TI - Structure and biosynthesis of fungal cell walls: methodological approaches. AB - Fungal cell walls possess a characteristic chemical composition differentiating fungal cells from other cell types. For this reason, the mechanisms involved in cell-wall formation represent a potential target for selective antifungal drugs. Understanding the structure and biosynthesis of fungal cell walls opens the ways for design of effective drugs for treating fungal diseases. This article reviews the history methods employed in chemical and structural analysis of fungal cell walls and in studies concerning their formation. PMID- 14533479 TI - Targeted disruption of Streptomyces globisporus lndF and lndL cyclase genes involved in landomycin E biosynthesis. AB - Streptomyces globisporus strains with knockouts in lndF and lndL genes, previously identified as possibly encoding cyclases governing cyclization of the nascent oligoketide ('polyketide') chain during the biosynthesis of the antitumor angucycline landomycin E, were prepared. On combining the results of sequence analysis and HPLC of extracts from mutant strains, lndL was suggested to control the first cyclization-aromatization event and lndF to be responsible for the 3rd 4th ring formation. PMID- 14533476 TI - Structure and functional properties of prokaryotic small noncoding RNAs. AB - Most biochemical, computational and genetic approaches to gene finding assume the Central Dogma and look for genes that make mRNA and have ORFs. These approaches essentially do not work for one class of genes--the noncoding RNA. In all living organisms RNA is involved in a number of essential cell processes. Functional analysis of genome sequences has largely ignored RNA genes and their structures. Different RNA species including rRNA, tRNA, mRNA and sRNA (small RNA) are important structural, transfer, informational, and regulatory molecules containing complex folded conformations that participate in recognition and catalytic processes. Noncoding RNAs play an number of important structural, catalytic and regulatory roles in the cell. The size of the sRNA genes ranges from 70 to 500 nucleotides. Several transcripts of these genes are processed by RNAases and their final products are smaller. The encoding genes are localized between two ORFs and do not overlap with ORFs on the complementary DNA strand. As aptamers, some sRNA bind small molecular components (metal ions, peptides and nucleotides). This review summarizes recent data on the functions of prokaryotic sRNAs and approaches to their identification. PMID- 14533480 TI - Characterization of the Streptomyces coelicolor A3(2) wblE gene, encoding a homologue of the sporulation transcription factor. AB - The gene corresponding to the recently identified whiB-paralogous gene wblE in S. coelicolor was found after sequencing the downstream region of the stress response sporulation-specific sigma-factor gene, sigH, in S. coelicolor A3(2). Sequence analysis has revealed an ORF exhibiting high similarity to sporulation transcription factors WhiB and WhiD. A stable null mutant of the wblE gene was obtained by integrative transformation, via double cross-over. Disruption of the S. coelicolor wblE gene appeared to have no obvious effect on growth, morphology, differentiation, and production of the pigmented antibiotics actinorhodin and undecylprodigiosin. Expression of the wblE gene was investigated during differentiation by S1 nuclease mapping, using RNA prepared from S. coelicolor A3(2) and its isogenic sigF and sigH mutants grown to various developmental stages. A single promoter was identified upstream of the wblE coding region. The wblEp promoter was induced at the beginning of aerial mycelium formation and its activity decreased later in differentiation. No differences in expression of the wblEp promoter were detected in S. coelicolor A3(2) mutants in sigF and sigH genes for sporulation-specific sigma factors. Sequence of the wblEp promoter showed partial similarity to the consensus sequence of the extracytoplasmic sigma factors. PMID- 14533482 TI - Viability of dried vegetative cells or filaments, survivability and/or reproduction under water and light stress, and following heat and UV exposure in some blue-green and green algae. AB - Vegetative cells in dried, mucilagenous mass of Gloeocapsa aeruginosa and Aphanothece nidulans, reticulum of Hydrodictyon reticulatum, mucilagenous mass of Chroococcus minor, and filaments of Oedogonium sp. and Scytonema hofmanni died within 1/2, 1/2, 1/2, 1, 3 and 6 h, respectively, while dried vegetative filaments of Phormidium foveolarum retained under similar storage conditions viability for 4 d. P. foveolarum tolerated 1 mol/L NaCl. The resistance to desiccation in P. foveolarum exhibited similar dependence as that to heat or UV light. The water stress imposed on growing algae either on high-agar solid media or in NaCl-containing liquid media reduced at various levels or altogether inhibited the survival of vegetative parts in all, the cell division in C. minor, G. aeruginosa and A. nidulans, formation of heterocyst and false branch in S. hofmanni, oogonium in Oedogonium sp., and daughter net in H. reticulatum. Heat or UV shock of any level also produced similar effects as that by water stress. P. foveolarum tolerated low light level of 10 and 2 mumol m-2 s-1 and no light longer than the rest of other algae studied. Tolerance of microalgal forms to water, heat or UV stress depends primarily upon cell-wall characteristics or cell sap osmotic properties rather than their habitats, morphology and prokaryotic or eukaryotic nature. PMID- 14533481 TI - Cross-resistance to strobilurin fungicides in mitochondrial and nuclear mutants of Saccharomyces cerevisiae. AB - In yeast the resistance to kresoxim-methyl and azoxystrobin, like the resistance to strobilurin A (mucidin) is under the control of both mitochondrial cob gene and the PDR network of nuclear genes involved in multidrug resistance. The mucidin-resistant mucl (G137R) and muc2 (L275S) mutants of Saccharomyces cerevisiae containing point mutations in mtDNA were found to be cross-resistant to kresoxim-methyl and azoxystrobin. Cross-resistance to all three strobilurin fungicides was also observed in yeast transformants containing gain-of-function mutations in the nuclear PDR3 gene. On the other hand, nuclear mutants containing disrupted chromosomal copies of the PDR1 and PDR3 genes or the PDR5 gene alone were hypersensitive to kresoxim-methyl, azoxystrobin and strobilurin A. The frequencies of spontaneous mutants selected for resistance either to kresoxim methyl, azoxystrobin or strobilurin A were similar and resulted from mutations both in mitochondrial and nuclear genes. The results indicate that resistance to strobilurin fungicides, differing in chemical structure and specific activity, can be caused by the same molecular mechanism involving changes in the structure of apocytochrome b and/or increased efflux of strobilurins from fungal cells. PMID- 14533483 TI - Polyphasic taxonomy of symbiotic rhizobia from wild leguminous plants growing in Egypt. AB - About 20 strains of rhizobia from wild legumes were characterized based on numerical analysis of phenotypic characteristics, nodulating ability, fatty acid methyl esters (FAME) and SDS-PAGE profiles of whole cell proteins. FAME analysis revealed that palmitic (16:0), stearic (18:0) and arachidonic (20:0) were detected in most of wild-legume rhizobia, the latter being uncommon in fatty acid profiles of Rhizobium and Sinorhizobium. Numerical analysis of FAME classified strains of wild-legume rhizobia into 9 clusters and one heterogeneous group. There was both agreement and disagreement with the clustering data based on phenotypic analysis and FAME analysis. Four strains were grouped together in the same cluster based on both methods. However, 4 another strains, which were placed in one cluster of phenotypic analysis, were distributed in several clusters after FAME analysis. SDS-PAGE of whole-cell proteins revealed that the rhizobial strains exhibited protein profiles with peptide bands ranging from 5-19 band per profile and showed molar mass of 110-183 kDa. As in the case of FAME analysis, numerical analysis of protein bands was compared with clustering of phenotypic analysis. Agreement of the two methods was obvious when clustering some strains but conflicted in the classification of some other strains. However, integration of the three methods could be the basis of a polyphasic taxonomy. The twenty strains of wild-legume rhizobia were finally classified as follows: 12 strains related to Rhizobium leguminosarum, 5 strains related to Sinorhizobium meliloti and 3 strains to Rhizobium spp. Rhizobia nodulating wild herb legumes are among indigenous strains nodulating crop legumes in cultivated as well as noncultivated lands. PMID- 14533484 TI - Detection of Burkholderia pseudomallei in rice fields with PCR-based technique. AB - Burkholderia pseudomallei Ara- in rice fields was detected using PCR-based techniques with 16S RNA and flagella gene primer sets. The sensitivity of these PCRs was at least 1 CFU/mL of B. pseudomallei Ara- preincubated into Ashdown's medium for 6 h. B. pseudomallei Ara- DNA from watery soil were more detectable than from dry soil. The distribution of this DNA was mainly found at a depth of 300-600 mm under crop-covered fields, but not detected in the location of soil close to the land surface. The results suggest that PCR based on 16S RNA and flagella gene primer sets can be applied to investigate the presence of B. pseudomallei Ara- in contaminated soil of rice fields. PMID- 14533485 TI - Decolorization and bioremediation of molasses wastewater by white-rot fungi in a semi-solid-state condition. AB - Molasses wastewater (vinasse; the by-product of distillation of fermented sugar) was decolorized and its chemical oxygen demand (COD) was reduced in static cultivation using the fungi Coriolus versicolor, Funalia trogii, Phanerochaete chrysosporium and Pleurotus pulmonarius ('Pleurotus sajorcaju'). The effect of cotton stalk on decolorizing and COD removing capability of four fungi was determined. In the entire concentration range tested (10-30%), wastewater was effectively decolorized by C. versicolor and F. trogii. Cotton stalk addition stimulated the decolorization activity of all fungi. The utilization of cotton stalk represents several advantages due to its function as an attachment place and as a source of nutrients; its use also reduces process costs. PMID- 14533486 TI - Lack of efflux mechanism in a clinical isolate of Pseudomonas aeruginosa highly resistant to beta-lactams and imipenem. AB - An isolate of Pseudomonas aeruginosa from cystic fibrosis was highly resistant to beta-lactams and beta-lactamase inhibitors. The resistant determinants of clinical isolate to imipenem, ceftazidim, cefriaxone and cefepime were conjugally nontransferable. The slow or nonenzymically mediated breakdown of imipenem and other broad-spectrum beta-lactams suggested the resistance of P. aeruginosa isolate to these drugs which may be attributed to both permeability and efflux. Impaired penetration of imipenem and other beta-lactams through the membrane was detected by a diminished expression of outer-membrane proteins of approximate molar mass of 46 and 39 kDa, matched to OprD and OprF, respectively. Efflux resistance mechanism for meropenem and beta-lactams has been ruled out since the isolate failed to express outer-membrane protein of approximately 50 kDa which is matched to the OprM protein channel. Thus, reduced permeability in the clinical isolate is the main mechanism conferring resistance against beta-lactams including imipenem. PMID- 14533487 TI - Changes in amount of bacteria during gut passage of leaf litter and during coprophagy in three species of Bibionidae (Diptera) larvae. AB - To elucidate the interaction between bacteria and saprophagous Diptera larvae, the amounts of bacteria in leaf litter, individual gut compartments, and feces of three species of Bibionidae (Bibio pomonae, Bibio marci, and Penthetria holosericea), feeding either directly on leaf litter or on fecal pellets produced from leaf litter by larvae of the same species, were assessed by determining total direct counts and viable counts on solid media at different pH. In P. holosericea, the effect of various cultivation temperatures on direct counts of bacteria in individual compartments was also demonstrated. In all species, the amount of bacteria in the anterior mesenteron was lower than in the consumed food, regardless of whether the larvae were feeding on leaf litter or feces, and increased again in the posterior part of the gut. The amount of bacteria in these compartments was generally higher in larvae feeding on feces than in those feeding on leaf litter, whereas the amount of bacteria found in the ceca varied. In B. marci, the amount of bacteria in the mesenteron sections able to grow on alkaline medium (pH 9) was higher than that of bacteria able to grow on slightly acidic medium (pH 5.5) during both the first and the second gut passage. In B. pomonae and P. holosericea, this increase was observed only during the second gut passage. The effect of gut passage in P. holosericea on changes in direct counts of bacteria was more pronounced when the larvae were fed at 5 degrees C as compared to 20 degrees C. Radiolabeled bacteria were digested in the gut and utilized as a source of energy and nutrients by the larvae; digested bacteria represented up to 10% of the material assimilated by the larvae. Lysozyme activity in whole-gut extracts of P. holosericea had a pH optimum of at pH 7, indicating a low in situ activity in the alkaline mesenteron. Proteinase activity, however, had an optimum at pH > 12, suggesting that the digestion of bacteria in the bibionid gut is caused by a combination of digestive proteinases and alkaline pH in the anterior mesenteron. PMID- 14533488 TI - Activation of human leukocytes by lipid A from E. coli strains adapted to quaternary ammonium salt and amine oxide. AB - The immunomodulatory activities of monophosphoryl lipid A (MLA) and diphosphoryl lipid A analogues obtained from the sensitive strain of E. coli and from the resistant strains adapted to a quaternary ammonium salt and an amine oxide were compared. All analogues considerably stimulated the activity of human leukocytes although the analogue from the sensitive strain at a higher concentration significantly suppressed phagocytosis. The MLA analogue exhibited a suppressive effect on the microbicidal activity of human leukocytes against E. coli and the peroxidase activity. Adaptation of bacteria to amphiphilic antimicrobial compounds, which is accompanied by chemical changes in their lipid A, only slightly reduced their immunomodulatory activity when compared with the analogue from the sensitive strain. On the other hand, the diphosphoryl analogues were less active than MLA. PMID- 14533489 TI - Elaboration of enzyme immunoassay based on recombinant antigens and intended for diagnostics of syphilis. AB - Enzyme immunoassay (EIA) using recombinant antigens for the detection of Treponema pallidum-specific antibodies in sera of syphilis patients was developed. Four low-molar-mass Treponema antigens (Tp15, Tp17, TmpA, Tp47) were investigated; 17- and 47-kDa proteins were demonstrated as immunodominant as they permitted to obtain the most sensitive EIA. Using a mixture of these proteins a 3rd-generation-EIA kit Dia-Syph was constructed, its sensitivity being 99.4% during tests of 165 sera of syphilitic patients. No false result was obtained on the commercial panel PSS01 (BBI, USA). The specificity of the elaborated test system (99.7%) was determined on 295 sera. PMID- 14533490 TI - The detection of anti-Candida antibodies on experimental animal models. AB - The levels of anti-Candida antibodies were determined in experimental animals immunized with 4 different yeast doses (0.3, 0.6, 1.2, 2.4 x 10(8) CFU) at weekly intervals. After immunization (sampling intervals 1, 2, and 3 months), the intravenous blood was examined for the presence of serum anti-Candida antibodies. After one month, the titers of anti-Candida antibodies reached 1:40-1: 1280 and remained at the same level after two months in the majority of animals; only in a few of them the titers increased or were detected de novo. After three months, when the animals were no longer immunized, a decreasing trend in antibody titers was detected in the majority of animals. PMID- 14533491 TI - [Delay in the diagnosis of tuberculosis]. AB - In five patients, a 46-year-old Surinamese man, a 45-year-old Dutch barkeeper, a 41-year-old woman from Eritrea, and an 18-year-old Afghan woman and her 43-year old mother, tuberculosis was diagnosed and treated after a considerable delay. Such a late diagnosis and subsequent treatment is not uncommon for tuberculosis as the symptoms, physical examination, laboratory tests, imaging techniques and bacteriological results are often not very specific. However under certain circumstances, tuberculosis (including the extrapulmonary manifestations) should be considered at an earlier stage, especially among certain risk groups such as the homeless, drug addicts, immigrants and asylum seekers. In the five patients discussed the causes of delay were identified as: delay caused by the asymptomatic phase of the disease, the patient's delay in presenting, a delay on the part of the physician, a delay in diagnostic confirmation and a delay in the treatment. PMID- 14533492 TI - [Breast cancer and the use of the hormones around the menopause]. AB - Following the publication of the 'Million women study', commotion has broken out in the media about the relationship between hormone use around the menopause and breast cancer. In this study, women were asked about possible risk factors for breast cancer, including hormone replacement therapy, prior to breast cancer screening. After an average follow-up period of 2.6 years, it was found that breast cancer more frequently occurred among women who had recently used an oestrogen-progestagen preparation (relative risk (RR): 2.00; 95%-CI: 1.88-2.12), an oestrogen preparation (RR: 1.30; 1.21-1.40) or tibolone (RR: 1.45; 1.25-1.68) than among women who had not used either of these. Methodologically, the study contains both strengths and weaknesses. However, the findings agree with those from an earlier randomised study. Current users of hormone use were more likely than never users to die from breast cancer (RR: 1.22; 1.00-1.48). For the prescription of hormone replacement therapy, the twice as high relative risk of breast cancer together with the other potentially life-threatening side effects of hormone replacement therapy need to be weighed up against the complaints which form the indication for the prescription. Therefore, the advice for hormone replacement therapy continues to be that it should, if necessary, be made available as a treatment option for a short period (3 months) and only in the case of serious and debilitating complaints. Long-term preventative use, as happens in practice, is not indicated due to the risk of relatively serious side effects. PMID- 14533493 TI - [Improved risk counselling possible for patients with unruptured intracranial aneurysms]. AB - The 'International Study of Unruptured Intracranial Aneurysms' (ISUIA) investigators have recently published the results of a large prospective, observational study on risks of rupture and risks of treatment in patients with unruptured intracranial aneurysms. In patients with aneurysms smaller than 7 mm no rupture of the carotid, middle cerebral and anterior communicating arteries occurred during follow up (mean 4.1 years) if there was no history of subarachnoid haemorrhage from a separate aneurysm. Risks were higher in aneurysms of the posterior circulation (2.5% in 5 years), and in patients with a previous episode of subarachnoid haemorrhage from another aneurysm (1.5 to 3.4% in 5 years for aneurysms < 7 mm). Size was the most important risk factor for rupture, and aneurysms of the posterior circulation > 25 mm had a 50% cumulative risk of rupture over 5 years. The absence of rupture in some subgroups, such as small aneurysms of the anterior communicating artery, is probably explained by the small numbers in these subgroups. Similarly, the lack of identification of risk factors other than size is probably explained by insufficient power of the study. The overall risk (defined as death or dependence) of surgical treatment was 13%, with patient age and size and site of the aneurysm as the most important risk factors. Endovascular treatment carried a 7 to 10% risk in most subgroups; only patients with aneurysms > 25 mm had increased risks. Age was not a risk factor for endovascular treatment. By making use of these results, patients can now be given sound advice regarding the benefits of preventive treatment in each individual case. PMID- 14533494 TI - [Research into new methods for diagnosing, treating and preventing tuberculosis]. AB - Tuberculosis control requires improved diagnostics, drugs, and vaccines. Their development is facilitated by progress in immunology, molecular biology, and genomics. In addition to sputum smear and culture, amplification techniques can already be used to diagnose tuberculosis and antigen-detection tests for this purpose are being developed. Molecular typing and DNA microarrays provide new insights in the natural history and transmission of tuberculosis. In addition to established drugs such as rifampicin, isoniazid, pyrazinamide, and ethambutol, a limited number of new drugs have been discovered such as rifampicin derivates and fluorochinolones. Improved screening techniques and insights from genomics may lead to new drugs being discovered. Factors hampering the development and evaluation of new vaccines include problems with extrapolation from animal models, incomplete natural immunity, and limited knowledge about protective immunity. However, new candidate vaccines are being developed and will be tested on humans in the near future. PMID- 14533496 TI - [Overweight and obesity; recommendations from the National Health Council]. AB - The Health Council of the Netherlands has issued a report on overweight and obesity. These are considered major threats to personal health, the public health care services and economic welfare. Overweight and obesity are no longer considered variations of the norm, but as a disease. As genetically predisposing factors cannot be influenced, major initiatives to contain further deterioration of the problem must be on environmental factors i.e. on macronutrient intake and physical exercise. Special target groups (young people, people of low socioeconomic status and immigrants) need to be defined. Government, health care organisations, industry and the general public should work together to combat overweight and obesity. The chances of success are not predictable but are certainly big enough to justify action. PMID- 14533495 TI - [Thrombosis prophylaxis in orthopaedic surgery: considerations and uncertainties]. AB - As it is not always possible to identify patients at extra risk of deep-venous thrombosis after major orthopaedic surgery, it is considered advisable to prescribe prophylactic treatment. Prophylaxis with a low dose of unfractionated heparin is more effective than no prophylaxis at all but less effective than low molecular-weight heparin or cumarine derivatives. Acetylsalicylic acid does not appear to have a place in the prevention of venous thromboemboli in elective total-hip and total-knee surgery. Its usefulness in the operative treatment of hip fractures has not been established. Starting prophylaxis postoperatively has not been shown to be less safe or effective than starting it preoperatively. The incidence of thrombosis is lower if the course of prophylactic treatment is lengthened to 4-6 weeks following arthroplasty of the hip or knee. The optimal period of medicinal prophylaxis has yet to be ascertained. Intermittent pneumatic compression appears to be effective after total-knee arthroplasty but in practice is difficult to deliver. Support stockings, a foot pump and rapid mobilization have little effect on the incidence of proximal deep-venous thrombosis. There are insufficient data regarding the use of thrombosis prophylaxis in day surgery. PMID- 14533497 TI - [Diagnostic image (157). A man with fever and a 'tache noire' after a holiday in South Africa. Rickettsiosis, probably African tick-bite fever]. AB - A 54-year-old man presented with fever and an eschar, probably caused by African tick-bite fever, contracted during a holiday in South Africa. He recovered rapidly after treatment with doxycyclin. PMID- 14533498 TI - [Hormone replacement therapy in the peri-menopausal and post menopausal period: more than half of the women were treated for more than one year]. AB - OBJECTIVE: To describe the use of hormone replacement therapy (HRT) and oral contraceptives (OC) among women in the Netherlands in the 45 to 69 years age group during the year 2002. DESIGN: Descriptive. METHOD: Pharmaceutical data on 42,032 women who were in the 45-69 years age group on 31 December 2002 was selected from the InterAction database, a collection of community pharmacy data from the north of the Netherlands. The duration of the therapy was investigated for women who had received at least one HRT or OC prescription in 2002. RESULTS: Oral contraceptives were used by 11.3% of women aged 50-54 years and 1.8% of women aged 55-59 years. The prevalence of HRT ranged from 6.0% to 11.7% depending on age. Of these users (n = 3885) 28.6% used an oestrogen-progestagen preparation. A treatment period of more than one year was found in 62.3% of women aged 45-49 years and 82.6% of women aged 55-59 years. In particular, preparations containing just oestrogen had been used for a longer period; 43.4% had used these for more than 5 years. CONCLUSION: More than half of the women who used HRT had done so for more than one year. PMID- 14533499 TI - [Thrombosis prevention in orthopaedic surgery: clinical practice in the Netherlands in 2002]. AB - OBJECTIVE: Establish the use of thromboprophylaxis in orthopaedic surgery both during and after the hospital admission. DESIGN: Cross-sectional study. METHOD: In April 2002, a letter was sent to all orthopaedic surgeons in the Netherlands announcing that at every hospital with a Department of Orthopaedic Surgery, an orthopaedic surgeon would be approached for a telephone survey. They were phoned in the months April-June 2002. This study included hospitals where major orthopaedic surgery (e.g. the insertion of hip or knee prostheses and hip fracture surgery) took place (n = 124) as well as clinics that only performed day treatments (n = 5). RESULTS: For major orthopaedic operations, 91% of the hospitals used low molecular weight heparin (LMWH) during the admission period: 36% as monotherapy and 55% in combination with coumarin derivates. In 85% of cases the use of LMWH was started preoperatively. Coumarin derivates were used as a monotherapy in 9% of the hospitals. In 37% of the hospitals the use of NSAIDs was continued, particularly in patients with rheumatoid arthritis. In 94% of the hospitals, the use of acetylsalicylic acid was always stopped. In 97% of the hospitals prophylaxis was given after discharge in the form of LMWH (37% of the cases) or coumarin derivates (63% of the cases). The use of thromboprophylaxis with respect to arthroscopies and plaster immobilisation was variable. PMID- 14533500 TI - [The waiting time for heart interventions: trends for percutaneous coronary interventions and cardiothoracic interventions]. AB - OBJECTIVE: (a) To describe trends in the number of heart interventions performed over time, (b) to determine the length of waiting lists for elective heart interventions in the Netherlands according to the monthly survey of the Supervisory Committee for Heart Interventions in the Netherlands [Begeleidingscommissie Hartinterventies Nederland (BHN)], (c) to compare the length of the waiting lists with existing standards, and (d) to determine the reliability of the waiting list survey. DESIGN: Prospective. METHOD: Data were obtained from the monthly waiting list survey of the 13 heart centres in the Netherlands (1 January 1999-30 November 2002) and from the intervention registry (1 January 1999-30 June 2001), which was complete for 10 centres. Both the survey and the maintenance of the registry are carried out by the Supervisory Committee for Heart Interventions in the Netherlands. RESULTS: (a) The number of percutaneous coronary interventions performed in the Netherlands has increased. The number of cardiothoracic interventions remained stable. (b) The number of patients waiting for a percutaneous coronary intervention is increasing by 16% per annum. In November 2002 there were 751 patients on the waiting list. The number of patients waiting for a cardiothoracic intervention increased by 20% per annum until August 2001 and since then there has been a decrease of 21% per annum. In November 2002, 1557 patients were on the waiting list. (c) The percentage of patients treated within existing standards has fallen to 78% for percutaneous coronary interventions and to 53% for cardiothoracic interventions. (d) The length of the waiting list and the waiting times obtained in the survey concurred with the data taken from the intervention registry. CONCLUSIONS: The length of the waiting list for heart interventions has increased and complies increasingly less with existing standards. The monthly waiting-list survey was a reliable method of determining the length of waiting lists for elective heart interventions. PMID- 14533501 TI - [Trigeminal trophic syndrome: 2 patients with a non-healing ulcer on the ala nasi]. AB - Two female patients, aged 74 and 91 years respectively, had a skin ulcer on a nostril and ipsilateral sensory paralysis in the area covered by the N. trigeminus. This is known as trigeminal trophic syndrome. It may be caused by (iatrogenic) damage to the trigeminal nerve by chronic manipulation of the senseless skin. Only in case of doubt, histological examination of a tissue sample is advised to exclude malignancy. The main treatment consists of instructing the patient not to manipulate the skin. In some patients surgical reconstruction is necessary. PMID- 14533502 TI - [Centenary of the Royal Dutch Central Association for the Prevention of Tuberculosis]. AB - The Royal Dutch Tuberculosis Association (Koninklijke Nederlandse Centrale Vereniging tot bestrijding der Tuberculose (KNCV)) was founded in 1903. Since then various interventions against tuberculosis have been introduced on the basis of medical and technological opportunities and in response to the epidemiological situation. The introduction of effective drugs during the 1940s and 1950s was by far the most important development and led to a sharp decline in the annual rate of infection and to the disappearance of tuberculosis as a common disease. Following the appointment of the independent National Policy Committee for Tuberculosis Control, Royal Dutch Tuberculosis Association could concentrate on its innovating tasks: the development of new interventions based on epidemiological data and scientific research. PMID- 14533503 TI - [The Dutch contribution to worldwide tuberculosis control: Royal Netherlands Association for the Prevention of Tuberculosis (KNCV) Tuberculosis Foundation]. AB - During the 1960s and 1970s tuberculosis was severely neglected in developing countries. Less than 50% of patients diagnosed were cured. However, an international breakthrough occurred in the 1980s with the excellent results from a short-course treatment: 80% cured. This was first achieved in Tanzania. Since 1993, this innovative approach has been known throughout the world as the 'directly observed treatment, short-course' (DOTS) strategy. Over the past 15 years the interest in international tuberculosis control has substantially improved. There has been a strong increase in the number of initiatives to rapidly extend the use of DOTS so that internationally set targets for tuberculosis control can be met (by 2005, 70% of infectious patients detected and 85% of these cured). The highest priority has been given to the 22 countries with the highest incidence of tuberculosis. There has been a significant increase in the number of countries implementing DOTS. The average rate of cure using DOTS is 80%. Since the 1970s the Royal Netherlands Tuberculosis Association (Koninklijke Nederlandse Centrale Vereniging tot Bestrijding der Tuberculose (KNCV)) has been involved in many international initiatives, such as the Stop TB Partnership, and it has participated in the development and implementation of policies for tuberculosis and tuberculosis-HIV control. However, there are still causes for concern: the number of tuberculosis patients is increasing in particular due to the tuberculosis-HIV co-epidemic and it is estimated that only 1 in 4 infectious tuberculosis patients were detected and treated in DOTS programmes during the year 2000. PMID- 14533504 TI - [The anti-synthetase syndrome: muscle disease and multisystem disorder at the same time]. PMID- 14533505 TI - [Withdrawal symptoms in a neonate following exposure to venlafaxine during pregnancy]. PMID- 14533506 TI - Ethics in action. "Fess up" to patients? PMID- 14533508 TI - When lymph edema takes hold. PMID- 14533509 TI - Defenses gone awry. Primary biliary cirrhosis. PMID- 14533510 TI - Spotting drug use. PMID- 14533511 TI - PA catheters. What the waveforms reveal. PMID- 14533512 TI - Do you suspect child abuse? PMID- 14533513 TI - Scribner made routine dialysis a reality. PMID- 14533514 TI - From here to eternity. How do physicians learn to let go? Part 2. Parting gifts what begins when technology ends. PMID- 14533515 TI - Common concerns among hospital-based dialysis providers. AB - Studies by Alan Collins, MD, and the U.S. Renal Data System have demonstrated that a proper balance of good dialysis therapies, including appropriate use of medications, leads to improved patient outcomes, including lower hospitalization rates and mortality. Additionally, this reduction in hospitalization and increase in patient longevity will improve the financial performance of both hospital based and freestanding dialysis providers. Good care for the patients is of financial benefit to the dialysis program for both hospital-based and freestanding dialysis facilities. This article highlights many of the challenges facing hospital-based dialyis providers, but it only scratches the surface. Hospital-based programs, while in the minority of dialysis providers, need to incorporate many of the lessons that the freestanding providers have learned. These lessons are not just modification of staffing models and the monitoring of productivity standards; the focus is on improving quality of care and capturing all of the revenue the program is entitled to receive. For hospital-based dialysis programs, improving patient outcomes also improves financial performance. PMID- 14533516 TI - NRAA develops group purchasing organization ... an idea whose time has come. PMID- 14533517 TI - Improving diabetes foot care in the dialysis facility. PMID- 14533518 TI - Patients educating patients. Let's add a little PEP to the renal community. PMID- 14533519 TI - Study looks at effectiveness of generics.. PMID- 14533520 TI - An ANNA invitational summit: nephrology nursing shortage and solutions. PMID- 14533521 TI - The nursing shortage and potential solutions: an overview. AB - Successfully solving the nephrology nursing shortage will require multiple strategies on national and local levels. Retaining current nephrology nurses is a key strategy. In addition, we, as a specialty, must help recruit people into nursing, and recruit student nurses and graduate nurses into the field of nephrology nursing. This article will describe the national nursing shortage and discuss how to create work environments that promote retention and satisfaction. PMID- 14533522 TI - Provider panel presentations. AB - To wrap up the three big items--attracting talent, the professional nurse role, and our structural disadvantages--I would offer the following take-away thoughts: We ought to do what we can to get our disproportionate share of RNs out of the health care system to work in dialysis. If we work together, we have a shot at making that happen. As an industry, we must exploit our strengths. We have to exploit the pride that comes in the services and the care that we provide. We need to create in a very visual way that difference for nurses who are considering which area of specialty to go into and where to create their careers. We, as providers, have a huge opportunity to create the right environment for our nurses. That's something we own. We have to fix it. We have to create a better place to work for nurses than any place else. We must create a more enriching place for nurses than anywhere else. That's up to us. I know a lot of us on the provider side are doing tons of things to make that happen and we need to keep driving that. We need a level playing field. We've proven the fact that we can deliver clinical outcomes with the best. Now we need help from our biggest customer to make sure we can compete for the resources that are necessary to sustain and improve those clinical outcomes on an ongoing basis. PMID- 14533523 TI - Renal hemodynamics: an overview. AB - The formation of urine is a process that begins with glomerular filtration and is greatly influenced by changes in renal hemodynamics. Selective filtration of the blood is possible because of the unique characteristics of the glomerulus and renal circulation. Many factors interact to maintain a consistent blood flow allowing filtration and urine formation to continue despite systemic changes in blood pressure. Factors that impact on renal hemodynamics include the autoregulatory mechanism, the renin-angiotensin mechanism, eicosanoids, kinins, the sympathetic nervous system (SNS), catecholamines, antidiuretic hormone, endothelin, nitric oxide, atrial natriuretic peptide, and dopamine. Knowledge of the effects of these factors will allow the nephrology nurse to predict, identify, and assist in the treatment of clinical conditions that can alter renal hemodynamics and glomerular filtration. PMID- 14533524 TI - Achieving optimal patient outcomes with intravenous iron. AB - Optimizing i.v. iron management is essential to the overall health of hemodialysis patients. Establishing an i.v. iron protocol, administering a safe and effective i.v. iron supplement, and developing a strong nurse/patient relationship are all indispensable to an effective and efficient hemodialysis program. PMID- 14533525 TI - Hemoglobin variability: impact on anemia management practices. AB - Data indicate that a wide range of factors can contribute to inter- and intrapatient variability in hemoglobin (Hb) levels in dialysis patients. Some of these factors are controllable and can be minimized by implementing nursing practices that encourage consistent and accurate Hb assessments and management. However, many of these factors are difficult or impossible to modify, and natural intrapatient variations are therefore expected and unavoidable. When natural variability leads to Hb levels that temporarily exceed 12 g/dL, appropriate protocol-guided nursing responses should include long-term, patient-specific laboratory trend analyses, data-driven adjustments in the anemia management prescription, and accurate nursing documentation of assessments, interventions, and outcomes. PMID- 14533526 TI - Inverse relationship between blood volume and blood pressure. AB - The outcome objectives for EG were met. In order for her blood pressure to improve, cardiac output had to improve. In her case, this could only be accomplished by reducing the vascular volume through aggressive ultrafiltration. Starling's Law and Laplace's Theorem provide the key to understanding this concept, namely that an increase or decrease in cardiac output will result in a corresponding increase or increase or decrease in mean arterial pressure (Smith & Kampine, 1990, p. 93). Thus, there is an inverse relationship between blood volume and blood pressure. In this case ultrafiltration treated the cause of hypotension. Discontinuing the ultrafiltration and/or giving a saline bolus would have been an incorrect attempt to treat the effects of hypotension. PMID- 14533527 TI - Readers respond to questions on heparin prescription and administration. PMID- 14533528 TI - A vascular access surveillance/monitoring program that can be instituted immediately. PMID- 14533529 TI - Diabetes update: the prevention and management of diabetes. PMID- 14533530 TI - Seeing visions become realities at kidney camp. PMID- 14533532 TI - Post-operative retention of urine in children. PMID- 14533533 TI - Nurse and mother: is there a conflict of interest? PMID- 14533534 TI - Adolescent facilities: a parent's view.... PMID- 14533535 TI - Adolescent facilities: the potential.... PMID- 14533536 TI - Child protection: every nurse's responsibility. PMID- 14533537 TI - Interprofessional learning and family-centred care. PMID- 14533538 TI - ISABEL: support with clinical decision making. PMID- 14533539 TI - Developing career opportunities through on-going education. PMID- 14533540 TI - Eating disorders: the role of hypnosis. PMID- 14533541 TI - Writing academically: using existing knowledge to improve writing style. PMID- 14533542 TI - The difference between incidence and prevalence. PMID- 14533543 TI - HAART and cardiology--current controversies and consequences. PMID- 14533544 TI - Evolution of the involvement of the cardiovascular system in HIV infection. PMID- 14533545 TI - Cardiovascular pathology in AIDS. PMID- 14533546 TI - Pathogenesis of HIV-associated cardiovascular disease. PMID- 14533547 TI - The pathogenesis of HIV-associated pulmonary hypertension. PMID- 14533548 TI - Pathogenesis of the HAART-associated metabolic syndrome. PMID- 14533549 TI - HIV-associated lipodystrophy: pathogenesis and clinical features. PMID- 14533550 TI - Assessment of autonomic and cardiovascular function in HIV disease. PMID- 14533551 TI - Atherosclerosis and HIV infection: diagnosis and treatment. PMID- 14533552 TI - Coronary heart disease in HIV-infected individuals. PMID- 14533553 TI - Coronary artery disease and stroke in HIV-infected patients: prevention and pharmacological therapy. PMID- 14533554 TI - Vasculitic syndromes in HIV-infected patients. PMID- 14533555 TI - HIV-associated pulmonary hypertension: diagnosis and treatment. PMID- 14533556 TI - HIV-associated cardiovascular complication in HIV-infected children. PMID- 14533557 TI - Cardiac surgery in the patient with human immunodeficiency virus. PMID- 14533558 TI - Guidelines for the prevention and management of cardiovascular complications in HIV-infected patients receiving HAART: The Pavia Consensus Statement. PMID- 14533559 TI - Interactions between antiretroviral agents and drugs commonly used to treat cardiovascular diseases according to the Pavia Consensus Statement. PMID- 14533560 TI - [Evidence-based evaluation of preventive procedures for osteoporosis, osteoporotic fractures and other diseases]. AB - In the present special articles entitled "Evidence-based prevention of osteoporosis and osteoporotic fractures", several preventive procedures for these diseases were evaluated for their effectiveness according to the best available evidence of literature. These procedures included optimization of calcium intake and physical activity as the primary prevention, bone density measurement as the screening, and prevention of falls in the elderly people. The present article described a method for the evaluation of preventive procedures for osteoporosis based on systematic review of the evidence, which was applicable for other preventive services of diseases. The next step to the review of evidence is how to communicate the evidence established to practitioners in the fields of preventive medicine and community health. One of the effective tools for this purpose is evidence-based preventive practice guideline. The Japanese Society for Hygiene should play a leading role in compiling a series of guidelines for the preventive procedures of various kinds of health problems such as lifestyle related diseases. PMID- 14533561 TI - [Optimization of calcium intake for the prevention of osteoporosis and osteoporotic fractures: a review of the evidence]. AB - One of the main focuses of lifestyle modification for the prevention of osteoporosis and osteoporotic fractures in Japan is improvement in dietary calcium intake. However, virtually no randomized controlled trial to assess the preventive effects of administration of calcium on the risk of fractures has been conducted in Japan. In this study, we reviewed all the scientific papers currently available from medical literature databases to propose evidence-based recommendations on the preventive procedures for osteoporosis. The result of the present systematic review gives the evidence showing that calcium supplementation or optimal dietary calcium intake increases bone density in childhood and adolescence and reduces the risk of fracture due to osteoporosis in the elderly people regardless of the gender. The evidence also supports the current health policy guiding the elderly to increase their dietary calcium intake in daily life. PMID- 14533562 TI - [Exercise and physical activities for the prevention of osteoporotic fractures: a review of the evidence]. AB - According that osteoporosis is the common condition in an aging society such as in Japan, much progress has been made in understanding the treatment and prevention of osteoporosis. Among potential risk factors, exercise and physical activities have been recognized as lifestyle factors that might influence the risk of osteoporosis and osteoporotic fractures. To assess the relationship between exercises including physical activities and the risk for low bone mass and osteoporosis-related fractures, a literature search over past 13 years was conducted. Accumulating evidence indicates that exercises decrease the risk for hip fractures among middle aged and older men and women. Exercises also help to maintain muscle strength, muscle volume, balance, and joint flexibility, which might prevent falls and fall-related fractures. One randomized controlled trial showed back-stretching exercise reduced the risk for vertebral fractures. The literature search also indicates that high-impact and/or weight-bearing exercise might increase the bone density in the elderly and the peak bone mass among young women, while there is no association between moderate or lower-impact exercise and bone mineral density. Future research should be required to evaluate the types and quantity of physical activity needed for the prevention of osteoporosis. PMID- 14533563 TI - [Effectiveness of screening for osteoporosis by bone density measurement for the prevention of fractures: a review of the evidence]. AB - To review evidence on the benefits of screening women and men for osteoporosis, a Pub Med search was performed in English papers published between 1990 and 2002. We used data from a cohort study to estimate risk of fracture from bone mineral density. Bone mineral density measured by dual X-ray absorptiometry (DXA) can predict bone fracture among elderly women, peri- and early post-menopausal women, and elderly men. It is recommended that all white women older than 65 years be screened routinely for osteoporosis. We suggest that Japanese elderly women should receive BMD measurements as a screening, but we have still issues to be solved including age from when the screening should be started, methods, and how to treat the women found to have osteoporosis at the screening. For peri- and postmenopausal women and elderly men, it might be beneficial to measure BMD as a screening and start treatment for those patients found to have osteoporosis. However, incidence of fractures for these people is lower than that for elderly women. One bone mass measurement can predict bone fracture risk for as long as over 10 years or more, but predictive ability of BMD decreases with time. Therefore, cost effectiveness needs to be reviewed to determine the benefits of screening among peri-menopausal women and men. Although bone assessment by quantitative ultra sound (QUS) method by ultrasound can also predict future fractures, only a relatively small number of longitudinal studies have been conducted in the Western countries, and there is no established evidence by means of longitudinal studies among Japanese. It is necessary in Japan to seek such evidence, however, since this method is widely used for an osteoporosis examinations. PMID- 14533564 TI - [Preventive strategy of fall for the prevention of fractures in older people: a review of the evidence]. AB - This review aims to examine the effectiveness of fall-related strategies for fracture prevention among elderly population. Previous studies were reviewed using PubMed and Japan Centra Revuo Medicina databases. Our five research questions concerned prediction of fracture using history of accidental falls, fracture risk management, and effectiveness of exercise programs, home modification and usage of hip protector. We used "accidental falls" and "fractures" as search strategies. Obtained results were as follows: a) History of falls predicts future hip fracture. b) Exercise programs including balance training have positive effects for fracture prevention. c) Fracture risk evaluation and management reduce the number of hip fractures among elderly population. d) Environmental hazard assessment and necessary home modification are effective in preventing fractures especially among elderly population with the history of falls. e) The hip protector is a beneficial device for the prevention of hip fractures among elderly people at high risk of falling. PMID- 14533566 TI - [The relationship of quality of life (QOL) with physical fitness, competence and stress response in elderly in Japan]. AB - OBJECTIVES: In order to evaluate the relationship of quality of life (QOL) with physical fitness, competence and stress response in the elderly population in Japan, a cross sectional field survey of elderly subjects was conducted. METHODS: This survey was taken in Naguri village, Saitama. The data collected included physical fitness, competence, stress response and QOL in addition to demographic variables. As for physical fitness indexes, grip strength (GS), single leg balance with eyes closed (SLB), bar grip ping reaction time (RT), trunk flexion (RF), ten-meter walking time (WT) and vital capacity (VC) were measured. The SF 36 was used for QOL assessment. RESULTS: A total of 120 elderly subjected participated to the survey. There were 42 males (73.5 +/- 5.74 years) and 78 females (74.2 +/- 6.17 years). The associations between physical health parameters in SF-36 and WT were highly significant: physical functioning (beta = 2.96, p < 0.001), role physical (beta = -3.64, p < 0.001), bodily pain (beta = 3.27, p < 0.001) and general health (beta = -3.14, p = 0.001). Psychological stress response had a negative correlation with social functioning (beta = -0.74, p = 0.024), role-emotional (beta = -2.34, p < 0.007) and mental health (beta = 0.97, p = 0.024) as determined by multiple regression analysis. The goodness-of fit indexes of the structural equation model describing the relationships among physical fitness, competence, stress response and QOL indicated excellent fit to the data with GFI = 0.95 and AGFI = 0.88. Stress response showed relatively stronger influence on QOL than physical fitness or competence. CONCLUSION: Although there were slight differences in degree of influence, physical fitness, stress response and competence were found to be clearly related to QOL in elderly subjects. To keep good QOL status, it is important to maintain good physical fitness and level of competence and to reduce stress response. PMID- 14533565 TI - [Characteristics of patients with intractable diseases receiving public financial aid for treatment--results of a nationwide survey, 1997]. AB - OBJECTIVES: In order to clarify the characteristics of medical institutions visited by patients with selected intractable diseases, we analyzed data from the fourth nationwide survey in 1997. METHODS: We asked 47 prefectural governments to provide data concerning every patient receiving financial aid for treatment of 39 selected intractable diseases from April 1997 to March 1998. Out of 399,719 whose information was reported by prefectural governments, we analysed data of 370,232 patients whose medical institutions were reported. We performed detailed analysis on the relation between patients' residences and locations of medical institution which the patients visited, and on the characteristics of medical institutions. These analyses were respectively compared by sex and age, the beginning year of the financial aid, whether the patients were inpatients or outpatients, type of insurance, and clinical division where the patient was treated. RESULTS: 1. Analysis showed that 7.4% of all patients were treated in medical institutions outside the prefectures where they lived. Patients who lived in the neighboring prefectures of huge cities like Tokyo, tended to be treated in the medical institutions there. 2. We found that 23.5% of patients were treated in university hospitals, and 11.9% were seen in clinics. 3. There was a difference between patients with SMON and patients with myastenia gravis, pemphigus, epidermolysis bullosa or primary pulmonary hypertension. Of the two groups, the former preferred to visit clinics and be treated in medical institutions located in the same cities, towns, and villages where they lived. On the contrary, patients with the latter 4 diseases tended to visit large hospitals and be treated in those outside their prefectures. 4. Elderly patients over 70 years old tended to be treated in clinics or hospitals located in their neighborhoods. 5. Compared with past surveys, the percentage of patients treated in university hospitals had decreased, and that of patients treated in clinics had increased year by year. No change was found in the proportion of patients treated in medical institutions outside their prefectures. CONCLUSIONS: By the present analysis of a nationwide survey taken in fiscal year 1997, we were able to clarify the characteristics of medical institutions visited by patients with selected intractable diseases. This kind of analysis should be continued to obtain important information on the epidemiology of intractable diseases. PMID- 14533567 TI - [Utilization of school lunch dietary data to estimate theoretical maximum daily intake (TMDI) of selected pesticide residues]. AB - OBJECTIVES: The theoretical maximum intake values of pesticides by body weight (TMI) were estimated for school children and the differences in district and age group were compared. Furthermore TMI values for children (7-11 years old) were calculated from Japanese National Nutrition Survey (NNS) data. Our TMI values for school children were compared with those from NNS data and evaluated to assess whether our values were appropriate for use in the calculation of theoretical maximum of daily intake (TMDI) of the pesticides. METHODS: The TMI values of pesticides for children were estimated using the intake amounts of agricultural products calculated from school lunch data in three elementary school districts (in Hyogo, Tokyo and Yamagata prefectures) and the survey of data from meals at home. Furthermore, TMI values for children (7-11 years old) were estimated from NNS data relating to agricultural products intake. Target pesticides for TMI calculations were chlorpyrifos, cyhalothrin, fenbutatin oxide, malathion, meltribuzin, oxamyl, permethrin and vamidothion. RESULTS: TMI of malathion from school lunch data corresponded to 58% of ADI and was too large for exposure from one meal in comparison to other pesticides (8-24%). Our estimations of TMI of chlorpyrifos, cyhalothrin, oxamyl and vamidothion in the school lunches were smaller than those from the NNS data. However TMI of malathion calculated from the school lunch data was larger than that from the NNS data. TMI of fenbutatin oxide, meltribuzin and permethrin from the school lunch data were similar to those from the NNS data. CONCLUSIONS: The TMI values for children in three districts depended upon the agricultural products used in school lunches. Even though the survey methods differed between the NNS's and our data, the TMI values closely corresponded. We concluded that the school lunch data might compensate for the lack of NNS data in order to estimate more appropriate TMDI for children's ingestion of agricultural products. PMID- 14533568 TI - [Association between the rates of low birth-weight and/or preterm infants and aircraft noise exposure]. AB - OBJECTIVES: Intense noise exposure having been observed in vicinal areas around the U.S. military airfields in Okinawa, Japan, suggests the possibility of adverse effects on fetal growth, as studies have reported such effects around other airfields. This study analyzes the birth records in Okinawa prefecture and investigates whether lower birth weights of infants and shorter gestation periods are observed around the airfields. METHODS: The records of 160,460 births in 15 municipalities around the Kadena and Futenma airfields from 1974 to 1993 were subjected to analysis. Average WECPNL among residents in each municipality was calculated as a measure of noise exposure, since the birth records did not contain information on precise birth addresses but only the municipalities. The odds ratios of low birth weight, i.e. under 2,500 grams, and preterm birth, i.e. less than 37 weeks, were obtained by multiple logistic regression analysis with adjustment for the primary factors that would be related to fetal growth. The factors included sex, maternal age, live birth order, occupation of householder, legitimacy of the infant, year of birth and interaction between maternal age and live birth order. RESULTS: The logistic regression analysis showed a significant dose-response relationship between low birth weight and noise exposure. The significance probability of trend test was less than 0.0001. The adjusted odds ratio was 1.3 in the highest noise exposure area, which was Kadena Town, located in the immediate vicinity of the Kadena airfield. Significantly higher rates of preterm births were also found across the noise exposed municipalities. Although the obtained results were not adjusted for some confounding factors like smoking habit, another survey conducted by the present authors did not show particular differences in the female smoking rate between Kadena Town and the other municipalities around the Kadena airfield. CONCLUSION: Aircraft noise exposure is most likely to cause adverse effects on fetal growth, raising the rates of low birth weight and preterm infants around the Kadena airfield. PMID- 14533569 TI - [Health and socio-economic condition of certified Minamata disease patients, 1999]. AB - OBJECTIVES: To clarify the needs and to consider establishing a social support system for patients with Minamata disease (MD), or methylmercury poisoning, by investigating their health and socioeconomic conditions. METHODS: The total number of people certified as having MD in May 1999 by the Kumamoto and Kagoshima Prefecture Government Committees on MD was 2265. We sent two questionnaires to 917 individuals who were surviving at that time, which corresponded to 40.5% of the total number of MD patients. The first survey sought information on the individual's health-seeking behavior, and the second survey was about their socio economic conditions and requirements for welfare and medical care in the future. RESULTS: The average age among male patients was 68.0 +/- 13.2 yrs (n = 477) and that among female patients was 71.2 +/- 13.0 yrs (n = 440). The response rates were 45.7% (n = 416) for the first questionnaire and 38.6% (n = 354) for the second questionnaire. Among the MD patients, 71.7% judged their health condition to be 'bad' or 'very bad', and 97.4% received medical treatments that included acupuncture or moxacautery and massage. Regarding the activity of daily living (ADL), which includes 'communicating', 'walking', 'eating', 'use of toilet', 'dressing' and 'taking a bath', the rates of 'independent' were relatively low among those under 49 yrs and those over 75 yrs compared with the other age groups. Many individuals emphasized that they had anxiety about their health and health care in the future. CONCLUSION: We concluded that the quality of life (QOL) of MD patients was low. It is important to consider developing a social support system for MD patients. PMID- 14533570 TI - Reporting breast implant complications. AB - The Food and Drug Administration (FDA) defines a complaint (or complication) as "any written, electronic, or oral communication that alleges deficiencies related to the identity, quality, durability, reliability, safety, effectiveness, or performance of a device after it is released for distribution." The FDA, the medical device manufacturers, and the health care providers using these devices share the responsibility for complaint reporting, investigation and follow-up. Each has unique yet interrelated roles and responsibilities in this process. PMID- 14533571 TI - Overcoming complications of breast implants. AB - Having nice breasts is important to many women and their self-images. For those women who have had congenital deformities, were not blessed naturally with the shape they wanted, have had experiences that have adversely affected breast tissue, or have had a mastectomy as a result of breast cancer, breast implants can provide a boost in self-confidence. This article provides an overview of breast implant issues and treatments, and reviews breast implant complications, their potential causes and treatments. PMID- 14533572 TI - BOTOX: a review. AB - BOTOX cosmetic is proving to be an affordable alternative for thousands of people looking to enhance their appearance without the cost, risks and downtime associated with surgical procedures. Botulinum toxin is a neuromuscular blocking agent produced by Clostridium Botulinum, an anaerobic bacterium. When first discovered BOTOX was pinpointed as the cause of severe paralysis acquired through the ingestion of contaminated food. Paralysis related to BOTOX is caused by chemodenervation, which is a result of blockage of the presynaptic release of acetylcholine at the neuromuscular junction. The corrugator (medial eyebrows), crow's feet, forehead, platysmal neckbands, and the jowl are frequently targeted areas for BOTOX injections. There are no current guidelines for physical limitations following the administration of BOTOX. Side effects associated with BOTOX infections appear to be either local, due to the paralysis of adjacent muscles, or mild flu-like symptoms lasting a few days in a small number of patients. PMID- 14533573 TI - AAAASF releases revised criteria. PMID- 14533574 TI - Mixing medicines, foods, herbs, and surgery: monoamine oxidase inhibitors (MAOIs). PMID- 14533575 TI - Health Insurance Portability and Accountability Act (HIPAA): protected health information (PHI)--physician's office challenges. PMID- 14533576 TI - No easy answers. PMID- 14533577 TI - Playing God. Moral tensions in health care. PMID- 14533578 TI - Moral integrity. Faithful living in a fallen world. PMID- 14533579 TI - Caught! Charting ahead. PMID- 14533580 TI - The ethic of care. A christian perspective. PMID- 14533581 TI - A listening heart. PMID- 14533582 TI - Human cloning. Investigating the issues. PMID- 14533583 TI - Medical futility. When is enough, enough? PMID- 14533584 TI - A view from the combat zone. PMID- 14533585 TI - War within & without. PMID- 14533586 TI - Navy nurse. A call to lay down my life. PMID- 14533587 TI - Remembering Vietnam. PMID- 14533588 TI - Pursuing peace in a world at war. PMID- 14533589 TI - Edith Cavell. WWI nurse, hero, martyr. PMID- 14533590 TI - Do you still need an annual Pap? PMID- 14533591 TI - Knee arthritis, part II: treatments that work. PMID- 14533592 TI - A guide for paying less for prescription drugs. PMID- 14533593 TI - Debugging your travel plans. PMID- 14533594 TI - Should I try using glucosamine to relieve the pain of arthritis? PMID- 14533595 TI - What causes toenail fungus and how can I get rid of it? PMID- 14533596 TI - Fetal respiratory activity and pulmonary hypoplasia. PMID- 14533597 TI - Fetal upper respiratory tract function in cases of antenatally diagnosed congenital diaphragmatic hernia: preliminary observations. AB - Fetal upper respiratory tract function was studied in five cases of antenatally diagnosed congenital diaphragmatic hernia and in 16 cases of uncomplicated pregnancy at gestational ages ranging from 27 to 38 weeks. The evaluation of fetal upper respiratory tract function was performed using ultrasonography combined with color-flow and spectral Doppler analysis. In all cases with uncomplicated pregnancy, fetal breathing-related nasal and oropharyngeal fluid flow was seen at the level of the nose. The five cases with congenital diaphragmatic hernia all demonstrated fetal breathing activity by thoracic wall movement. In four of the fetuses, perinasal fluid flow was seen by the Doppler technique. The fetus with no demonstrated perinasal flow during breathing movements died in the early neonatal period and had pulmonary hypoplasia. Observation of the fetal breathing-related nasal and oropharyngeal fluid flow in cases of antenatally diagnosed congenital diaphragmatic hernia provides a rationale to hypothesize that the absence of this phenomenon is a useful marker for prenatal prediction of pulmonary hypoplasia. PMID- 14533598 TI - Fetal echocardiography and color Doppler flow imaging: the Rotterdam experience. AB - The adjunctive role of Doppler color flow mapping in the evaluation of cardiac structures and function was studied in 440 normal fetuses between 17 and 22 weeks of gestation (median, 20 weeks) and in 73 fetuses with suspected congenital heart disease between 16 and 38 weeks of gestation (median, 28 weeks). Flow through atrioventricular and arterial valves was generally easy to identify and identification was successful in approximately 90% of the normal fetuses. Flow in the pulmonary veins and through the foramen ovale was visualized in approximately 60% of normal cases. In the group with suspected congenital heart disease, Doppler color flow imaging provided additional information on both cardiac structure and function in 34 fetuses, on function alone in 13 fetuses and on structure alone in 20 fetuses. No additional information was collected in six fetuses. PMID- 14533599 TI - Measurement of limb circumferences with three-dimensional ultrasound for fetal weight estimation. AB - A large number of formulae have been published for fetal weight estimation. This study examines the possibility of using three-dimensional ultrasound to measure the limb circumference. This technology provides us with the opportunity to visualize two simultaneous, orthogonal planes, allowing the exact determination of the transverse plane at the mid-point of the limb. A total of 157 patients were scanned in the week prior to delivery. The following variables were measured in 154 fetuses: biparietal diameter, head circumference, transverse abdominal diameter, abdominal circumference, femur length, thigh circumference and arm circumference. The whole population was split into three subgroups according to the abdominal circumference centile (< 10th centile, 10-90th centile, and > 90th centile), because this single variable has the strongest correlation with neonatal weight (R = 0.901). With forward stepwise multiple regression analysis, four models were elaborated, one for the whole population and one for each subgroup. The small-for-dates fetuses need a targeted formula including only two variables, thigh circumference and femur length. Our data suggest that the thigh circumference measurement improves the estimation for growth-retarded fetuses and that the arm circumference improves the weight prediction for the others. The measurement of the two variables, made possible with three-dimensional ultrasound, has improved our results in fetal weight estimation. PMID- 14533600 TI - The sensitivity and specificity of subjective and a semi-quantitative technique of amniotic fluid volume assessment in predicting intrapartum morbidity. AB - The aim of this study was to evaluate the ability of two techniques of amniotic fluid volume assessment to predict intrapartum morbidity in a high-risk population. A total of 213 pregnant women at the Grace Maternity Hospital, Vancouver, Canada were examined ultrasonically and the amniotic fluid volume was assessed subjectively and using the amniotic fluid index. Both techniques showed high specificity and negative predictive values for all outcome measures, but had an associated poor sensitivity and positive predictive value. Both techniques were similar in their ability to identify situations involving intrapartum morbidity. It is concluded that amniotic fluid assessment is a weak predictor of intrapartum morbidity but may be useful in certain conditions such as when the pregnancy is post-dates. PMID- 14533601 TI - Preoperative staging of breast cancer by palpation, mammography and high resolution ultrasound. AB - In breast cancer preoperative determination of the tumor size is important for planning breast-conserving operations. In 100 patients with breast cancer, the preoperative tumor size was measured using clinical, mammographic and sonographic examinations and correlated with the results of a subsequent histological examination. Using a high-resolution real-time system, 98 tumors were visible. It was possible to detect not only early tumors under 1 cm in diameter, but also intraductal tumor components. This contributed greatly to the accuracy of the diagnosis. The sonographic measurement of tumor size demonstrated a correlation coefficient of 0.91 and was thus superior to mammography (0.79) and palpation (0.77). Measurement of the total tumor spread, including 39 multicentric lesions, showed an overestimation of 5% for the mammographic measurements and an overestimation of 4% for the sonographic measurements. Tumor extension was underestimated in 33% of the mammograms but in only 3% using ultrasound examination. The results, along with those of other studies, highlight the role of sonography in the diagnosis of breast cancer. PMID- 14533602 TI - Echocontrast agent enhanced color flow imaging of the breast. AB - An ultrasound contrast agent which survives transit through the pulmonary circulation after injection into a peripheral vein can also be used for the demonstration of flow characteristics in the peripheral organs. Such a substance (SH U 508 A, Schering AG, Berlin) was tested in the Gynecological Department of the University of Marburg for its suitability for enhancing color-coded Doppler signals in mammary diagnosis in a group of ten women with breast tumors. The observations made are compared with the histologically confirmed diagnoses. The very distinct enhancement of the color-coded Doppler signals allows reliable demonstration of the vascularization characteristics, not only of the malignant, but also of all benign solid tumors examined. Color signals were even recorded from normal mammary tissue, which means that the demonstration of perfusion can no longer be used on its own as a diagnostic criterion. Observation of the characteristics of the blood supply to a tumor might, on the other hand, be an advantage, not only in respect of tumor typing, but also as regards the prognosis of a tumor. In this connection, the registration of the arrival, retention and wash-out phases specific to a contrast agent appears to furnish particularly interesting information about the growth dynamics of a tumor. PMID- 14533603 TI - Stage I ovarian cancer by transvaginal color Doppler sonography: a report of 18 cases. AB - A total of 18 cases with ovarian carcinoma stage I (15 stage Ia and three stage Ib) were studied retrospectively to evaluate the efficiency of transvaginal color Doppler sonography in detecting the disease in its early stages. Four asymptomatic women (two cases with morphologically normal ovaries, and two with simple unilocular cysts) were found during the screening program. These cases would have been missed without the use of transvaginal color Doppler sonography. One ovary with stage Ia, and another ovary in a case with stage Ib ovarian cancer were missed. These data show the ability of transvaginal color Doppler sonography to detect ovarian cancer as early as stage I even in asymptomatic women as well as in the morphologically normal ovary. PMID- 14533604 TI - Where is the "E" in MCH? PMID- 14533605 TI - Parents' needs after ultrasound diagnosis of a fetal malformation: an empirical deficit analysis. AB - We studied 56 pregnant women and 24 partners after ultrasound examination in the 18-24h gestational week revealed a fetal malformation. The subjects were followed through the process of examination, information sharing and counselling about the option of terminating the pregnancy. Regardless of sociodemographic variables or attitude towards the pregnancy, the diagnosis was always traumatic. A detailed critique of the physician's behavior and function emphasized his important role in the process of coming to terms with the malformation and gave clear indications of the positive expectations parents had. The affected patients expected the role of the professionally competent expert to be combined with that of the empathetic approachable counsellor, whose ste-by-step explanation would allow them to decide for themselves whether to terminate the pregnancy. The partner's participant was an essential aid to adjustment. It remains to be seen to what extent the functions of the doctor are realistic and justified. PMID- 14533606 TI - From the editors. PMID- 14533607 TI - Toward a pluralistic account of parenthood. AB - What is it that makes someone a parent? Many writers--call them 'monists'--claim that parenthood is grounded solely in one essential feature that is both necessary and sufficient for someone's being a parent. We reject not only monism but also 'necessity' views, in which some specific feature is necessary but not also sufficient for parenthood. Our argument supports what we call 'pluralism', the view that any one of several kinds of relationship is sufficient for parenthood. We begin by challenging monistic versions of gestationalism, the view that gestation uniquely grounds parenthood. Monistic and necessity gestationalism are implausible. First, we raise the 'paternity problem'--necessity gestationalists lack an adequate account of how men become fathers. Second, the positive arguments that necessity gestationalists give are not compelling. However, although gestation may not be a necessary condition for parenthood, there is not good reason to think that it is sufficient. After further rebutting an 'intentionalist' account of parenthood, in which having and acting on intentions to procreate and rear is necessary for parenthood, we end by sketching a pluralistic picture of the nature of parenthood, rooted in causation, on which gestation, direct genetic derivation, extended custody, and even, sometimes, intentions, may be individually sufficient for parenthood. PMID- 14533608 TI - Reasoning in bioethics. AB - It is striking that some arguments in the bioethical literature seem implausible, counterintuitive, and even ridiculous when reported to competent moral agents. When examined, these arguments bear uncanny resemblances to the discourse of patients with debilitating mental disorders. I examine the kinds of irrationality involved, and discuss the fact that such irrationality is worrying in a discipline that purports to serve as a guide for real-life practical reasoning. I offer some thoughts about correctives that we might use to temper some of the odd opinions that bedevil our subject in the name of ethical analysis. It seems that one ought to be suspicious of neatly rational arguments that produce counterintuitive conclusions, but the alternative seems to be that we explore new constructions of old problems in Bioethics such that our discussion of them does justice to what we regard as of fundamental significance to our lives together as human beings. PMID- 14533609 TI - Anorexia nervosa and refusal of naso-gastric treatment: a response to Heather Draper. AB - Imposing artificial feeding on people with anorexia nervosa may be unethical. This seems to be Heather Draper's suggestion in her article, 'Anorexia Nervosa and Respecting a Refusal of Life-Prolonging Therapy: A Limited Justification.' Although this is an important point, I shall show that the arguments supporting this point are flawed. Draper should have made a brave claim: she should have claimed that people with anorexia nervosa, who competently decide not to be artificially fed, should be respected because everybody is entitled to exercise their autonomy, not only 'in the middle' of their life, but also at the end of it, or when their own life is at stake, because autonomy also extends to the most difficult moments of our life, and, ultimately, 'stretches [...] far out into the distance' at the end of it. I explain why Draper should have made the brave claim, and why she has not made it. I conclude that a defence of people's entitlement to competently refuse artificial feeding cannot rest upon the arguments developed by Draper. Whether or not we should respect competent refusal of artificial feeding depends on the normative strength that we are ready to ascribe to the principle of autonomy, to the moral relevance that we ascribe to the circumstances in which a person's autonomy is exercised, and, perhaps, eventually, on our sense of compassion. PMID- 14533610 TI - "I do not care for a lunatic's role": modes of regulation and resistance inside the Colquitz Mental Home, British Columbia, 1919-33. AB - This article chronicles the experiences of 100 male patients who were confined at the Provincial Mental Home, Colquitz, Vancouver Island between 1919 and 1933. Through an analysis of clinical records and other sources,I reconstruct the organizational and human environment that prevailed at Colquitz during the tenure of its first lay supervisor, Granby Farrant; I chart the diverse modes of discipline through which officials sought to maintain the institutional regimen; and I document the efforts undertaken by patients to resist authority, transcend their surroundings, and seek redemption and hope both inside and beyond the Colquitz wall. PMID- 14533611 TI - [The sick patients of Dr. Tissot (1728-1797)]. AB - Samuel Auguste Tissot (1728-1797) was a Swiss physician from Lausanne, who enjoyed a great reputation in his lifetime, thanks to his publications and professional expertise. His personal records contain many pieces of correspondence and more than a thousand written consultations, which are documents describing the disease of a person, that were sent to a physician in order to solicit advice on diagnosis and treatments. This collection is very interesting in many aspects, notably for giving an insight into the patients' points of view. This perspective, which is often difficult to adopt because of the lack of archival materials, has been presented recently as an essential alternative to the traditional history of medicine. Written consultations allow an original reflection on the perception or expression of sickness and shed new light on the patient-practitioner relationship. They reveal a complex network of mediations, that is, actions undertaken by individuals at any step of the epistolary consultations. This article aims at defining the different kinds or levels of mediations that one can find in the documents in order to understand when and why there are sometimes mediators, and which role they play. Such a study should demonstrate that the experience of sickness is not only the business of a single person, but involves several people. The patient-doctor relationship is not exclusively a binary one, but often includes the active participation of third parties. PMID- 14533612 TI - "A state bordering on insanity"?: identifying drug addiction in nineteenth century Canadian asylums. AB - This article examines the growing awareness of drug addiction as form of mental illness in several Canadian lunatic asylums in the last half of the nineteenth century and the beginning of the twentieth. Whereas in the 1870s and 1880s, medical and reform associations formed to cure and treat addiction and inebriety, asylum evidence suggests that it was not until the turn of the century that drug habituation was considered a condition which merited admission to asylum. Prior to the turn of the century, drug use appeared in the psychological profile of asylum entrants only as an attendant condition of a more traditional form of mental illness, such as mania or melancholia. Asylum physicians, seeking traditional categories, and utilizing subjective classification methods, generally would not consider addiction to be a distinct mental illness. At the end of the century, shifts in diagnostic convention and the official endorsement of those shifts signalled a change that was taking place in the asylum. The impact of drug addiction on the psychological profile of a patient was attracting more attention in the asylum. Subsequently drug addiction joined other earlier causes of mental illness, such as masturbation, and also began to be recognized as a mental condition worthy of treatment at the public asylum. Its status as mental disease proper, however, remained a point of debate. PMID- 14533613 TI - [Epidemic disease and regulation of migration in Canada]. AB - Beginning with the French Revolution and the Napoleonic Wars, the Parliament of Lower Canada adopted a rational and scientific mechanism for regulating migratory movements within its territory. Other than internal imperatives pertaining to security, this mechanism revolved around a prophylactic control of these movements of population. With the great migrations of 1830-1834 and 1847, the politicians of Lower Canada and the United Canada subsequently needed to revisit their regulatory mechanism in order to render it more efficient. In truth, new arrivals were frequently the spreaders of infectious diseases. Yet more importantly, the great epidemics, such as that of cholera in 1832, or of typhus in 1847, were not merely physically lethal, but were potentially disastrous to the social order. How then to resolve the potential pathological threats to the physical body with those to the political body? The political powers of the day adopted an entire method of thinking and a vocabulary which was more scientific, replete with medical references. Adopted under the seal of an emergency, these new measures of state control of prophylaxis were greatly inspired by those medical theories concerning infection and contagion which were prevalent prior to the arrival of the Pasteurian model of medicine. Those in power then used this new framework to analyze the question as a rational political problem. In the end, they would establish a tighter mechanism of state-controlled prophylaxis through the law, through regulations, and through devices for surveillance and control such as quarantine. The guarantor of social discipline through the erosion of individual freedom, this mechanism of state-run prophylaxis of migratory movements would, from that point forward, be justified by the health demands of the Cite organique, of the public health. PMID- 14533614 TI - Anorexia nervosa and refusal of naso-gastric treatment: a reply to Simona Giordano. PMID- 14533615 TI - Ezetimibe for lowering blood cholesterol. AB - Ezetimibe is the first drug in a new class called cholesterol absorption inhibitors. There is no evidence that ezetimibe will reduce rates of death or hospitalization. When it is taken alone or when it is added to existing statin therapy, ezetimibe can reduce serum cholesterol in patients who have an increased risk of future coronary artery disease. The safety and tolerability of ezetimibe alone or combined with a statin has not been established in trials beyond 12 weeks. PMID- 14533616 TI - Maternalism and the homeopathic mission in late-Victorian Montreal. AB - This article examines the interactions between homeopathy and maternal values in the nineteenth century through a case study of Montreal's homeopathic community and the Montreal Homeopathic Hospital, which opened in 1894. Through an analysis of women's diverse roles within the institution, it demonstrates the variety of ways in which maternalism found expression. It argues that the maternal ideology was particularly compatible with homeopathy's self-proclaimed mission to the poor, the emphasis it placed upon inculcating familial values within the homeopathic community, its appeal to women as patients, its claim to reduce child mortality, and the image it cultivated of the special role of the homeopathic mother. PMID- 14533617 TI - [The moral lessons of miasma: the "Memorandum on cholera" of Joseph-Charles Tache]. AB - In the spring of 1866, the Canadian government feared that a cholera epidemic was imminent. A conference of medical experts was held at Ottawa to determine the nature of the disease on the basis of "authentic facts" but the participants were unable to agree among themselves. Faced with the necessity of advising Canadians on how to live in time of plague, and yet unable to define clearly the nature of cholera, in his Memorandum on Cholera the deputy-minister of Agriculture, J.-C. Tache, translated medical preoccupations into matters of the government of oneself and others, in terms heavily accented by his fundamentalist Catholic religious beliefs. PMID- 14533618 TI - [Deaths at an early age in the city of Quebec, 17th-19th centuries]. AB - Archaeological excavations carried out in two ancient cemeteries (1657-1844) of Notre-Dame-de-Quebec Basilica in old Quebec City allowed us to analyze the skeletal remains of more than 175 children who died before they were 10 years old. The proportion of infants is higher than expected according to the parish registers presumably because archaeologists recovered the remains of infants whose death had not been recorded in the registers. Two peaks of mortality have been noted: among the newborns and between one and two years old. The observed bone lesions lead us to believe that prenatal stress and rickets might be responsible for these high mortality rates. Although the samples from the two cemeteries were contemporaneous and drawn from the same community, there are some significant differences from the morbidity and mortality standpoint, tentatively attributed to the fact that most of the children recovered from one of the cemeteries were the victims of an epidemic while those from the other cemetery died in normal circumstances. This study shows how the analysis of skeletal remains can shed light on pathological conditions which otherwise would remain undocumented and how paleoanthropology opens new perspectives of research on diseases and death in the past. PMID- 14533619 TI - Pedestrian roadway fatalities. PMID- 14533620 TI - Major ongoing stroke trials. PMID- 14533621 TI - Prevention & women's health: making the health care system more responsive to women. Grantmakers In Health. AB - This report is the first in a series of programs made possible by a grant to Grantmakers In Health (GIH) from The Commonwealth Fund to bring grantmakers together with experts in policy, practice, and research to exchange information and ideas about key health issues facing the nation. On May 10, 1999, GIH convened a small group of grantmakers along with experts in community health, research, and government for an Issue Dialogue on prevention and women's health. Intended to both share lessons learned and stimulate new ideas, the Issue Dialogue proved to be a lively and insightful forum on the challenges and opportunities in funding projects in this area. This Issue Brief brings together key points from the day's discussion with factual information on women's health and grantmaker activities drawn from a background paper prepared for Dialogue participants. PMID- 14533622 TI - Bioethics after the Enlightenment. PMID- 14533623 TI - Coping with managed care: responding to the needs of the elderly. Grantmakers In Health. AB - This report is the second in a series of programs made possible by a grant to Grantmakers In Health (GIH) from The Commonwealth Fund to bring grantmakers together with experts in policy, practice, and research to exchange information and ideas about key health issues facing the nation. On June 17, 1999, GIH convened a small group of grantmakers along with officials from the Health Care Financing Administration and other experts in consumer education, managed care, and health services research for an Issue Dialogue on managed care and the elderly. Intended to both share lessons learned and stimulate new ideas, the Issue Dialogue proved to be a lively and insightful forum on what health funders can do to support development of consumer information, tools, and program models for this vulnerable population. This Issue Brief brings together key points from the day's discussion with factual information on managed care within the Medicare program and grantmaker activities drawn from a background paper prepared for Dialogue participants. PMID- 14533624 TI - [Folic acid isn't sufficient for prevention of congenital defects. Report from the 42nd Teratologic Society Conference--6/22-27, 2002--Scottsdale Az, USA]. PMID- 14533625 TI - Youth mentoring: a social development approach to youth health promotion. Grantmakers In Health. AB - On November 3, 1999, GIH convened a forum, co-sponsored with Grantmakers for Children, Youth and Families (GCYF), featuring grantmakers who have made major commitments to youth mentoring as a health promotion strategy as well as researchers and those operating programs in the field. Participants engaged in a lively and open exchange of ideas, experiences, and information about what grantmakers can do to support and improve such programs. This Issue Brief brings together keypoints from the day's discussion with factual information on youth mentoring and grantmaker activities drawn from a background paper prepared for Dialogue participants. PMID- 14533626 TI - Reflections on (the age of) thresholding: a commentary on the (coming) age of thresholding. PMID- 14533627 TI - The threshold of bio-ethics: philosophical warrant in the thought of Stephen Erickson. PMID- 14533628 TI - Bioethics for thresholders: a brief introduction. PMID- 14533629 TI - The new age, Christianity, and bioethics. PMID- 14533630 TI - The (coming) age of thresholding. A concluding unscientific postscript. PMID- 14533631 TI - Searching for the truly human: standing at the precipice of a post-Christian age. PMID- 14533632 TI - Bioethics and the coming age of thresholding. PMID- 14533633 TI - The new eugenics and the newborn: the cousinage of eugenics and infanticide. PMID- 14533634 TI - Catholic reflections on the human genome. PMID- 14533635 TI - The wrongs of animal rights. PMID- 14533636 TI - Femoral blockade for fractured neck of femur in the emergency department. PMID- 14533637 TI - Gestating the embryos of others. Surrogacy? Adoption? Rescue? PMID- 14533638 TI - Multiple doses of activated charcoal: time for reappraisal II. PMID- 14533639 TI - Aquinas on euthanasia, suffering, and palliative care. PMID- 14533640 TI - Special crash investigations. PMID- 14533641 TI - Precocious signs of polycystic ovaries in young girls. PMID- 14533642 TI - Prevnar's cost creates treatment disparity. PMID- 14533643 TI - Flu treatment for the elderly makes economic sense, study says. PMID- 14533644 TI - New technique lowers computed tomography radiation dose for children. PMID- 14533645 TI - Dedicated trauma service improves emergency department triage time, lowers mortality. PMID- 14533646 TI - Current awareness in prenatal diagnosis. PMID- 14533647 TI - Current awareness in prenatal diagnosis. PMID- 14533648 TI - Home test measures total cholesterol. PMID- 14533649 TI - [Human disease genomics: the main topic of post-genomic era]. PMID- 14533650 TI - Black walnut. PMID- 14533651 TI - Bioaerosols and the scientific method. PMID- 14533652 TI - Outcomes to measure the effectiveness of asthma treatment. PMID- 14533653 TI - Toxic mold: phantom risk vs science. AB - OBJECTIVE: To review the available literature on the subject of fungi (molds) and their potential impact on health and to segregate information that has scientific validity from information that is yet unproved and controversial. DATA SOURCES: This review represents a synthesis of the available literature in this area with the authors' collective experience with many patients presenting with complaints of mold-related illness. STUDY SELECTION: Pertinent scientific investigation on toxic mold issues and previously published reviews on this and related subjects that met the educational objectives were critically reviewed. RESULTS: Indoor mold growth is variable, and its discovery in a building does not necessarily mean occupants have been exposed. Human response to fungal antigens may induce IgE or IgG antibodies that connote prior exposure but not necessarily a symptomatic state. Mold-related disease has been discussed in the framework of noncontroversial and controversial disorders. CONCLUSIONS: When mold-related symptoms occur, they are likely the result of transient irritation, allergy, or infection. Building-related illness due to mycotoxicosis has never been proved in the medical literature. Prompt remediation of water-damaged material and infrastructure repair should be the primary response to fungal contamination in buildings. PMID- 14533654 TI - Intolerance to dietary biogenic amines: a review. AB - OBJECTIVE: To evaluate the scientific evidence for purported intolerance to dietary biogenic amines. DATA SOURCES: MEDLINE was searched for articles in the English language published between January 1966 and August 2001. The keyword biogenic amin* was combined with hypersens*, allerg*, intoler*, and adverse. Additionally, the keywords histamine, tyramine, and phenylethylamine were combined with headache, migraine, urticaria, oral challenge, and oral provocation. Articles were also selected from references in relevant literature. STUDY SELECTION: Only oral challenge studies in susceptible patients were considered. Studies with positive results (ie, studies in which an effect was reported) were only eligible when a randomized, double-blind, placebo-controlled design was used. Eligible positive result studies were further evaluated according to a number of scientific criteria. Studies with negative results (ie, studies in which no effect was reported) were examined for factors in their design or methods that could be responsible for a false-negative outcome. Results of methodologically weak or flawed studies were considered inconclusive. RESULTS: A total of 13 oral challenge studies (5 with positive results and 8 with negative results) were found. Three of them (all with positive results) were considered ineligible. By further evaluation of the 10 eligible studies, 6 were considered inconclusive. The 4 conclusive studies all reported negative results. One conclusive study showed no relation between biogenic amines in red wine and wine intolerance. Two conclusive studies found no effect of tyramine on migraine. One conclusive study demonstrated no relation between the amount of phenylethylamine in chocolate and headache attacks in individuals with headache. CONCLUSIONS: The current scientific literature shows no relation between the oral ingestion of biogenic amines and food intolerance reactions. There is therefore no scientific basis for dietary recommendations concerning biogenic amines in such patients. PMID- 14533655 TI - Recurrent staphylococcal infections and chronic dermatitis in a 45-year-old man. PMID- 14533656 TI - Resource costs for asthma-related care among pediatric patients in managed care. AB - BACKGROUND: In 1998, the economic burden of asthma in the United States was estimated to be 12.7 billion dollars. Yet few studies have examined the relationship between the total costs of asthma-related care and measures of asthma morbidity. Understanding the relationship between total costs of asthma related care and morbidity can assist in designing the most cost-effective asthma care strategies to improve patient outcomes and minimize total costs. OBJECTIVE: To investigate correlates of asthma costs for children with mild-to-moderate persistent asthma and, specifically, to characterize how closely the percentage of predicted forced expiratory volume in 1 second (FEV1) and symptom days were correlated with costs of illness. METHODS: A total of 638 parents and children with mild-to-moderate persistent asthma in 4 managed care delivery systems in 3 different US geographic regions were enrolled. Symptom burden and annual resource utilization were determined from reports of physician visits, hospitalizations, emergency department visits, medication use, and parental missed workdays. Spirometry was conducted on children who were 5 years and older. To characterize the relationship between symptom days and the percentage of predicted FEV1 with costs, we specified a multivariate regression model. RESULTS: The median total annual asthma-related cost for the group was 564 dollars (interquartile range [IQR], 131 dollars-1602 dollars). Indirect costs represented 54.6% of total costs. Medicines accounted for 52.6% of direct costs. The mean percentage of predicted FEV1 was 101.6% (range, 39.3%-183.5%; IQR, 91.6%-111.3%), with 91.4% of patients with a percentage of predicted FEV1 of more than 80%. Based on multivariate modeling, increasing asthma severity, use of peak expiratory flow rate meters, younger age, low-income status and nonwhite race, and longer duration of asthma were significantly associated with increasing cost. Symptom days (P < 0.001) predicted annual costs better than percentage of predicted FEV1 (P < 0.16) in this group of children. CONCLUSIONS: For the large number of children with mild-to-moderate persistent asthma and normal or near-normal lung function, symptom days are predictive of health care costs. For these insured children receiving care from 3 large managed care providers, low-income status and nonwhite race were the strongest correlates for increased asthma-related costs. PMID- 14533657 TI - Histamine skin test reactivity following single and multiple doses of azelastine nasal spray in patients with seasonal allergic rhinitis. AB - OBJECTIVE: To determine whether azelastine nasal spray suppresses the dermal response to epicutaneous histamine in allergic patients and the duration of suppression after azelastine use is discontinued. METHODS: Seventy-eight patients with seasonal allergic rhinitis were entered into this randomized, double-blind, parallel-group, placebo-controlled study. Patients received either azelastine nasal spray (2 sprays per nostril twice daily) or placebo nasal spray for 14 days. Skin tests were performed 5 hours after the first dose of study drugs to determine the effect of a single dose of azelastine nasal spray on the wheal-and flare response to histamine. At the end of the 14-day treatment period, skin tests were performed 5 hours after the last dose of study drugs and at 24-hour intervals thereafter, until each patient's wheal-and-flare response to histamine (1.0 and 5.0 mg/mL) returned to within 20% of baseline values. RESULTS: A single dose of azelastine nasal spray did not significantly alter the wheal-and-flare response to histamine. The wheal response was within 20% of the baseline value in 82% and 88% (1.0 and 5.0 mg/mL of histamine, respectively) of the patients 5 hours after discontinuing 14 days of treatment with azelastine nasal spray. Wheal responses were within 20% of baseline values 48 hours after treatment was discontinued, whereas flare responses returned to within 20% of baseline within 48 hours in 92% of the patients. CONCLUSIONS: Azelastine nasal spray should be discontinued for at least 48 hours before beginning allergy skin test procedures. PMID- 14533658 TI - Efficacy and safety of fluticasone propionate 44 microg/salmeterol 21 microg administered in a hydrofluoroalkane metered-dose inhaler as an initial asthma maintenance treatment. AB - BACKGROUND: We wanted to evaluate whether treatment with an inhaled corticosteroid and an inhaled long-acting beta2-agonist is more effective than an inhaled corticosteroid alone for patients using as-needed albuterol who are initiating maintenance treatment. OBJECTIVE: To compare the efficacy and safety of twice-daily fluticasone propionate (FP) 88 microg and salmeterol 42 microg combined in a chlorofluorocarbon (CFC)-free (hydrofluoroalkane 134a) metered-dose inhaler (MDI) with the individual agents alone, each delivered through an MDI containing CFC propellants, in patient with persistent asthma previously uncontrolled with as-needed short-acting beta2-agonists alone. METHODS: Patients with asthma (n = 283) were randomized to twice-daily treatment for 12 weeks with FP 88 microg combined with salmeterol 42 microg (FSC) in a CFC-free MDI or the individual components alone from CFC-containing MDIs. RESULTS: At endpoint, mean change from baseline in morning predose forced expiratory volume in 1 second was significantly (P < or = 0.016) greater with FSC (0.69 L) compared with FP (0.51 L) or salmeterol (0.47 L). Fewer patients treated with FSC withdrew due to worsening asthma (1%) compared with FP (3%) or salmeterol (8%; P = 0.024). FSC significantly increased (P < or = 0.002) morning and evening peak expiratory flow rate at endpoint (66.5 and 51.5 L/min, respectively) compared with FP (43.0 and 29.9 L/min, respectively) and salmeterol (29.2 and 21.6 L/min, respectively). In addition, asthma symptom scores were reduced, and percentages of days with no asthma symptoms increased in all treatment groups. CONCLUSIONS: Treatment with FSC in a CFC-free MDI is more effective than FP or salmeterol alone in asthma patients who are symptomatic taking short-acting beta2-agonists alone. PMID- 14533659 TI - Elevations of local leukotriene C4 levels during viral upper respiratory tract infections. AB - BACKGROUND: One potential mechanism by which respiratory viruses trigger illness and complications is via the local elaboration of inflammatory mediators. OBJECTIVE: To determine whether there is an increase in local leukotriene C4 (LTC4) levels during experimental infection with influenza A virus (FLU), rhinovirus (RV), or respiratory syncytial virus (RSV). METHODS: Healthy adults were intranasally inoculated with a safety-tested strain of FLU (n = 29), RV (n = 16), or RSV (n = 21). Nasal lavage samples were collected, symptoms were recorded, and expelled nasal secretions were weighed before and then daily after challenge. Lavage samples were submitted for viral culture and assayed for LTC4 levels by radioimmunoassay. Serum antibody titers to the challenge viruses were assayed at baseline and 21 days after challenge. RESULTS: All subjects were infected as evidenced by viral shedding and/or seroconversion. Following infection, significant increases (P < 0.05 by analysis of variance) in LTC4 levels were measured for each virus. Furthermore, there was a temporal association between the local LTC4 levels and the development of illness. CONCLUSIONS: The results of this study, which used an adult experimental model, demonstrate elevations in locally produced LTC4 during respiratory infection with FLU, RV, and RSV. Future studies using antileukotriene agents may help elucidate the precise role of leukotrienes in mediating disease expression. PMID- 14533660 TI - Allergic rhinitis and polymorphisms of the interleukin 1 gene complex. AB - BACKGROUND: Allergic rhinitis is a chronic inflammatory disease with a genetic background. Inflammatory reactions are regulated by cytokines. Cytokine genes are polymorphic and have been implicated as candidate genes in allergy. OBJECTIVES: To study the significance of the interleukin 1 (IL-1) gene complex in allergic rhinitis. METHODS: Population-based, cross-sectional study. We studied the polymorphisms of 3 IL-1 gene complex genes, IL1A (+4845G>T), IL1B (-511 degrees C>T), and IL1RN (variable number of tandem repeats; IVS2, 86 bp, duplicates 2 to 5), in patients with allergic rhinitis. The study group consisted of 405 nonasthmatic individuals of whom 56 had allergic rhinitis. RESULTS: The genotype distribution differed significantly in all cytokine genes studied between subjects with and without allergic rhinitis. The difference was mainly due to an increased number of IL1A allele G homozygotes (67.9% vs 43.2%; odds ratio [OR], 2.8; 95% confidence interval [CI], 1.5-5.1), IL1B heterozygotes (72.2% vs 47.4%; OR, 2.8; 95% CI, 1.5-5.3), and IL1RN allele 2 homozygotes (18.5% vs 7.5%; OR, 2.8; 95% CI, 1.3-6.2) in allergic rhinitis. Haplotype analysis revealed a significant difference in the distribution of IL-1 gene complex haplotypes between subjects with and without allergic rhinitis (P = 0.005, 10 df). CONCLUSIONS: The IL-1 gene complex polymorphism is strongly associated with allergic rhinitis in nonasthmatic individuals. PMID- 14533661 TI - IgE-mediated allergy to fungal allergens in Finland with special reference to Alternaria alternata and Cladosporium herbarum. AB - BACKGROUND: Alternaria alternata and Cladosporium herbarum are common fungi in outdoor environments, but their clinical significance has not been elucidated in Finland. OBJECTIVE: To evaluate the prevalence of IgE-mediated allergy and clinical outcomes caused by sensitization to fungal allergens in patients with suspected allergy. METHODS: Skin prick tests (SPTs) were performed with C. herbarum in 6,376 patients and also with A. alternata in 1,504 of these patients. SPTs were repeated in 40 patients who showed a positive reaction to either allergen using commercial and in-house extracts. The association of SPT with allergen-specific IgE antibodies in serum was evaluated. Seven patients also underwent a conjunctival challenge test with these fungal allergens. RESULTS: The prevalence of positive SPT results to A. alternata and C. herbarum was low (2.8% and 2.7%, respectively). Among the 40 patients, atopic eczema/dermatitis syndrome was found in 58%, asthma in 44%, and rhinitis in 31%. Most of the patients displayed SPT reactions also to several other fungal allergens, and 75% to 80% showed a positive SPT reaction to allergens of pet animals or pollens. Four patients had a positive reaction to A. alternata and 6 to C. herbarum in the conjunctival challenge test. CONCLUSION: In the Finnish population with allergic symptoms, IgE-mediated sensitization to 2 common fungal allergens was rare and of minor clinical importance. SPT reactions to fungi are mostly observed in patients with multiple sensitivity to various allergens. PMID- 14533662 TI - Epidemiology of Japanese cedar pollinosis throughout Japan. AB - BACKGROUND: Japanese cedar pollinosis (JCP) is a common disease posing a major public health problem in Japan. For health care policy planning and development of new treatment modalities, investigation of the accurate prevalence and current status of JCP nationwide is imperative. OBJECTIVE: To ascertain the prevalence and the current status of JCP in Japan with use of a cross-sectional random sampling method. METHODS: In a nationwide survey conducted shortly after the peak pollen season, self-evaluation questionnaire were mailed to 10,920 subjects from 390 of 3,370 places in 12 regions in Japan. RESULTS: The response rate was 53.7%, and the usable response rate was 51.5%. The age-adjusted prevalence was 19.4%; the estimated prevalence was 13.1% after adjustment for misdiagnosis, incorrect answers, response rate, and case mix. JCP was most prevalent in the Kanto, Tokai, and Kinki areas and in working adults rather than in children or subjects ages 60 to 79 years. Total pollen count during the pollen season correlated well with the prevalence of JCP in individual regions. In subjects with JCP, nasal symptom were more severe than eye symptoms, 62.5% had severe or moderate interference with daily activities and consulted physicians, 54.1% took prescribed drugs, and 82% used some method to avoid pollen. CONCLUSIONS: The age-adjusted prevalence of JCP was 19.4% of the Japanese population, and estimated prevalence after correction of possible biases was 13.1%. Prescribed drugs treated approximately 60% of subjects with JCP, and 80% of subjects tried self-care by avoiding pollen. PMID- 14533663 TI - Absence of association of peripheral blood eosinophilia or increased eosinophil cationic protein with bronchial hyperresponsiveness during asthma remission. AB - BACKGROUND: The mechanisms responsible for persistent bronchial hyperresponsiveness (BHR) in adolescents with long-term asthma remission are poorly understood. OBJECTIVE: To determine whether BHR in adolescents with asthma remission is associated with peripheral blood eosinophilia, increased serum levels of eosinophil cationic protein (ECP), or both findings. METHODS: We classified 51 adolescents with long-term asthma remission (neither asthma-related symptoms nor medication during the previous 2 years) into 28 BHR-positive patients (methacholine PC20 [provocative concentration causing a 20% decrease in forced expiratory volume in 1 second] <18 mg/mL) and 23 BHR-negative patients. The peripheral blood eosinophil counts and serum ECP concentrations were compared between these 2 groups. Twenty-eight patients with symptomatic asthma (symptomatic group), matched for methacholine PC20 level with study subjects in the BHR-positive remission group, and 28 healthy adolescents (control group) were also studied. RESULTS: No significant differences in the peripheral blood eosinophil counts (262.1 +/- 117.0/microL vs 253.9 +/- 165.0/microL) and the serum ECP levels (15.6 +/- 10.0 microg/L vs 15.8 +/- 11.9 microg/L) were found between the BHR-positive and BHR-negative remission groups, respectively. The BHR positive remission group differed from the symptomatic group (372.9 +/- 190.3/microL, P < 0.05; 26.6 +/- 11.3 microg/L, P < 0.01) in both blood indices but resembled the control group (214.6 +/- 118.6/microL and 12.1 +/- 4.8 microg/L; both, no significant difference). CONCLUSIONS: BHR in adolescents with long-term asthma remission is not associated with peripheral blood eosinophilia or an increase in serum ECP concentration. This finding suggests that the mechanism underlying BHR in this clinical setting may differ from that in symptomatic asthma. PMID- 14533664 TI - Clinical characteristics of melon (Cucumis melo) allergy. AB - BACKGROUND: Although melon is a frequent allergy-eliciting fruit, allergic reactions to melon have rarely been reported. OBJECTIVE: To evaluate and describe the clinical characteristics of melon allergy in melon-allergic patients. MATERIALS AND METHODS: We evaluated patients allergic to melon and a control group of patients allergic to pollen. The diagnosis of melon allergy was based on a convincing clinical history, positive skin test results (prick-by-prick test), and positive results on oral challenge tests to melon. RESULTS: A total of 161 patients were included in the study: 66 in the melon allergy group and 95 in the pollen control group. The melon allergy group included 35 female and 31 male patients with a mean age of 26.6 +/- 2.7 years (range, 5-61 years). Although all patients had oral symptoms, 13 (19.7%) of the patients had extraoral symptoms and none experienced generalized urticaria or anaphylaxis. Excluding other Cucurbitaceae fruits, peach, fig, and kiwi most frequently elicited positive skin test results and symptoms. Up to 23% of melon-allergic patients had a concomitant latex sensitization. Melon allergy was especially linked to pollen allergy, since all the melon-allergic patients were also allergic to pollen. Some differential features with respect to the pollen allergy control group were a higher prevalence of asthma (odds ratio [OR], 2.13; P < 0.05) and a statistical increase in the frequency of sensitization to several tree and weed pollens, including Ulmus (OR, 42.8) and Ambrosia (OR, 22.4). CONCLUSION: The most important conditions linked to melon allergy are pollen allergy (100%), allergy to other nonrelated fruits, mainly peach (up to 62%), and latex sensitivity (up to 23%). Some differential features of the pollinosis in melon allergy were a higher prevalence of asthma and a higher frequency of sensitization to several weed and tree pollens. PMID- 14533665 TI - Comparisons of the complementary effect on exhaled nitric oxide of salmeterol vs montelukast in asthmatic children taking regular inhaled budesonide. AB - BACKGROUND: Inhaled, long-acting beta2-agonists or antileukotrienes are alternatives as add-on therapy for asthmatic children taking regular inhaled steroids. Any complementary effects would be relevant to the choice between these alternatives. Exhaled nitric oxide (FeNO) may reflect these effects. OBJECTIVE: To compare the control of FeNO provided by salmeterol or montelukast add-on therapy in asthmatic children undergoing regular maintenance treatment with a daily dose of 400 microg of budesonide. METHODS: The study included children with increased FeNO despite regular treatment with budesonide, 400 microg/d, and normal lung function. Montelukast, 5 mg/d, salmeterol, 50 microg twice daily, or placebo was compared as add-on therapy to budesonide, 400 microg, in a randomized, double-blind, double-dummy, crossover study. RESULTS: Twenty-two children completed the trial. The geometric mean FeNO level was 20 ppb (95% confidence interval [CI], 15-27 ppb) after salmeterol, which was significantly higher than after montelukast (mean, 15 ppb; 95% CI, 11-18 ppb; P = 0.002) and placebo (mean, 15 ppb; 95% CI, 10-21 ppb; P = 0.03). There was no difference in FeNO between the montelukast and placebo groups. Mean forced expiratory volume in 1 second (FEV1) was significantly increased after salmeterol (mean, 2.63 L; 95% CI, 2.34-2.91 L) compared with placebo (mean, 2.48 L; 95% CI, 2.19-2.77 L). Montelukast (mean, 2.57 L; 95% CI, 2.33-2.80 L) was no different than placebo. CONCLUSIONS: The FeNO levels were significantly higher after salmeterol add-on treatment compared with both placebo and montelukast add-on treatment. Salmeterol significantly improved lung function (FEV1) compared with placebo and nonsignificantly compared with montelukast. Montelukast failed to reduce FeNO and improve lung function compared with placebo in this group of children taking regular budesonide, 400 microg. PMID- 14533666 TI - Hepatic metabisulfite sensitivity in a patient with sclerosing cholangitis. AB - BACKGROUND: Primary sclerosing cholangitis is an uncommon chronic cholestatic liver disease with a poor prognosis in symptomatic cases. Genetic and immunologic alterations have been identified, and many possible etiologies have been entertained. Most treatments have limited benefit, and primary sclerosing cholangitis is a common cause for liver transplantation. OBJECTIVE: To describe a patient with documented primary sclerosing cholangitis associated with chronic ulcerative colitis, who developed hepatic toxicity following ingestion of metabisulfite. RESULTS: A placebo-controlled oral challenge suggested metabisulfite hypersensitivity with liver toxicity. He was treated with cobalamin (to prevent sulfite toxicity), low-sulfite diet, steroids, and antibiotics and has had an unusually benign course for 19 years. CONCLUSIONS: The hypersensitivity to oral metabisulfite in our patient appeared to be a significant trigger to flare-ups of his disease. Controlling the response to metabisulfite (along with recurrent antibiotic and steroid therapy) may have contributed significantly to the remarkably good outcome in this patient. PMID- 14533668 TI - Presidential address 2002. Parasitology in the 21st century. AB - During the past several weeks, I, like many of my predecessors, have read many speeches previously delivered at the annual meeting of the American Society of Parasitologists (ASP). These are wonderful, exciting papers, and I recommend them to you. The history of our Society and much of parasitological research and philosophy are contained in these speeches. An understanding of parasitology and specifically of the ASP in the 21st century necessitates an understanding of our roots. Just as proteomics, genomics, DNA, and Mendel are a continuum, so is our progression as a science and as a Society. This thought is not original with me. Several presidents have discussed this point in detail: Eloise Cram in 1956, "Stepping Stones in the History of the American Society of Parasitologists" (Cram, 1956); Martin Ulmer in 1978, "What's Past is Prologue" (Ulmer, 1978); Harry Hoogstraal in 1984, "ASP: Its Historic Role and Modern Opportunities" (Hoogstraal, 1985); and Mike Kemp in 1988, "Parasitology a Degenerate Discipline, Populated by Degenerate Scientists, Studying Degenerate Organisms?" (Kemp, 1989). PMID- 14533667 TI - Acute workup of vocal cord dysfunction. PMID- 14533669 TI - Morphology of the spores of Myxosoma cerebralis (Hofer, 1903) and M. cartilaginis (Hoffman, Putz, and Dunbar, 1965) [classical article]. PMID- 14533670 TI - Whirling disease of salmonid fish: life cycle, biology, and disease. AB - Myxobolus cerebralis is the myxozoan parasite responsible for causing whirling disease in salmonid fish. Although the parasite was first described nearly 100 yr ago, it received relatively little attention until the discovery of its 2-host life cycle in the mid 1980s. This was the first, complete, myxozoan life cycle to be described, and it was greeted with some skepticism because it united 2 stages of M. cerebralis that were previously classified in 2 separate taxa. In the last decade, there has been a renewed interest in this parasite because whirling disease has been implicated in the decline of wild trout populations in several western states in the United States. Subsequent research efforts have dramatically increased the understanding of the biology of M. cerebralis and the numerous factors that affect the severity of whirling disease in salmonid hosts. These efforts also have provided a great deal of new information concerning interactions between M. cerebralis and its aquatic oligochaete host Tubifex tubifex. This review examines the current state of M. cerebralis in relation to 3 categories: the life cycle, the salmonid hosts, and the oligochaete host. PMID- 14533671 TI - Field evidence for density-dependent effects in the trematode Microphallus papillorobustus in its manipulated host, Gammarus insensibilis. AB - Numerous studies have demonstrated that parasites with complex life cycles frequently manipulate the phenotype of their hosts to increase their transmission rate. Little is known, however, concerning density-dependent processes within infrapopulations of manipulative parasites--whether parasites cooperate to manipulate the host, whether competition counteracts with these potential cooperative benefits, or both. Here we explored these ideas, focusing on the association between the manipulative trematode Microphallus papillorobustus and its second intermediate host, the gammarid Gammarus insensibilis. From the data collected in the field, we found no evidence that co-occurring M. papillorobustus individuals benefit from the presence of conspecifics; instead, individuals in larger infrapopulations suffered reduced size and fecundity. Thus, the net effect of increasing density suggests that competition rather than cooperation is the dominant force in infrapopulations of M. papillorobustus. PMID- 14533672 TI - Helminth community structure of sympatric eastern American toad, Bufo americanus americanus, northern leopard frog, Rana pipiens, and blue-spotted salamander, Ambystoma laterale, from southeastern Wisconsin. AB - One-hundred twelve amphibians, including 51 blue-spotted salamanders, Ambystoma laterale, 30 eastern American toads, Bufo americanus americanus, and 31 northern leopard frogs, Rana pipiens, were collected during April-October 1996 from Waukesha County, Wisconsin and examined for helminth parasites. The helminth compound community of this amphibian assemblage consisted of at least 10 species: 9 in American toads, 8 in leopard frogs, and 3 in blue-spotted salamanders. American toads shared 7 species with leopard frogs, and 2 species occurred in all 3 host species. Although there was a high degree of helminth species overlap among these sympatric amphibians, statistically significant differences were found among host species and percent of indirect or direct-life cycle parasites of amphibian species individual component communities (chi2 = 1,015, P < 0.001). American toads had a higher relative abundance of nematodes, 59%, than larval cestodes, 31%, and larval and adult trematodes, 10%, whereas leopard frogs had a higher relative abundance of larval cestodes, 71.3%, and larval and adult trematodes, 25.3%, than nematodes 3.4%. This is related to ecological differences in habitat and dietary preferences between these 2 anuran species. Helminth communities of blue-spotted salamanders were depauperate and were dominated by larval trematodes, 94%, and few nematodes, 6%. Low helminth species richness in this host species is related to this salamander's relatively small host body size, smaller gape size, lower vagility, and more fossorial habitat preference than the other 2 anuran species. Adult leopard frogs and toads had significantly higher mean helminth species richness than metamorphs, but there was no significant difference in mean helminth species richness among adult and metamorph blue-spotted salamanders. Considering adult helminths, the low species richness and low vagility of caudatans as compared with anurans suggest that local factors may be more important in structuring caudatan helminth communities of salamanders than of anuran hosts. Helminth species infecting salamanders may be more clumped in their geographic distribution as compared with anurans, and the role of other hosts and their parasites at the compound community level may be important in structuring helminth communities of salamanders. PMID- 14533673 TI - Host sex preferences and transmission success by the water mite Unionicola foili (Acari: Unionicolidae) parasitic on the midge Chironomus tentans (Diptera: Chironomidae). AB - This study examined whether ecoparasitic larval Unionicola foili exhibited a sex bias when infecting laboratory populations of the host insect Chironomus tentans and whether an association with male or female midges increased the likelihood of larval mites returning to the aquatic habitat. When laboratory populations of C. tentans were exposed to larval U. foili, there was a higher prevalence of mites among female hosts at emergence (17 of 30 males vs. 25 of 30 females infected by mites). However, there was no significant difference in the distribution or abundance of larvae among infected male (mean = 2.3 larvae per host) and female (mean = 2.6 larvae per host) midges. Larval mites parasitizing both male and female chironomids were more likely to return to water than could be expected by chance. Mite larvae infesting female C. tentans were more likely to return to water when female hosts deposited egg masses in water, suggesting that oviposition plays an important role in cueing larvae parasitizing female midges to detach. The mechanism responsible for increasing the likelihood that mites parasitizing male hosts return to water remains unclear. Future studies will address the possibility of parasite-mediated changes in host behavior. PMID- 14533674 TI - Random spatial distribution of Schistosoma mansoni and hookworm infections among school children within a single village. AB - Schistosomes and soil-transmitted helminths currently infect a third of the world's human population. An important feature of these parasitic infections is their focal distribution, which has significant implications for control. Only a few studies have been carried out at the microepidemiological scale, comparing infection levels among individuals or households within a single village. In this study, data are presented from a cross-sectional survey, examining all children attending a primary school in rural Cote d'Ivoire over several consecutive days for Schistosoma mansoni, soil-transmitted helminths, and intestinal protozoa. All houses in the main village were mapped, and school children were linked to these households for small-area spatial analyses. Comparison between the 260 school children who live within the main village and the 89 children who reside in nearby settlements revealed significant differences in the overall prevalence and intensity of infections with S. mansoni and hookworm, confirming the focal nature of these 2 parasites. On the other hand, S. mansoni and hookworm infections exhibited random spatial patterns within the main village. The validity of these results is discussed in the context of this epidemiological setting, drawing attention to the issue of scale. Our findings have direct implications for intervention because they call for a uniform, community-wide approach to control schistosomiasis and soil-transmitted helminthiasis. Implementation can be relatively straightforward, and the proposed control approach might be cost effective and prove sustainable. PMID- 14533675 TI - Bots (Diptera: Oestridae) infesting a neotropical forest rodent, Proechimys semispinosus (Rodentia: Echimyidae), in Panama. AB - Botfly larvae (Cuterebra sp.) infesting spiny rats (Proechimys semispinosus) in 8 small islands in the Panama Canal were studied. Rats were live trapped monthly on each island from January 1991 through February 2000 and visually examined for the presence of bots. Overall, bot prevalence was 4.6% and differed statistically among island rat populations. Rats were simultaneously infested by as many as 4 bots. Overall bot intensity was 1.3 bots per infested rat and did not differ among islands. Mean bot density across all islands was 0.0111 and was greater during the dry seasons than during the rainy seasons, but it did not differ among islands. Bots were found during all the 12 calendar months, suggesting a multivoltine reproductive schedule. Although bot activity varied seasonally, there was little synchrony of bot activity among islands. Bot density was related negatively to rainfall but was not related to host density, suggesting that drier ambient conditions may promote reproduction by adult bot flies in this system. PMID- 14533676 TI - Neotropical Monogenoidea. 43. Diplectanum monticellii n. sp. (Diplectanidae) from the gills of Cynoscion leiarchus (Perciformes: Sciaenidae) in Brazil. AB - Diplectanum monticellii n. sp. is described from the gills of Cynoscion leiarchus, a marine Sciaenidae, from Itacuruca, Rio de Janeiro (type locality); Baia de Guaratuba, Parana; and Pontal do Sul, Parana in Brazil. The new species is characterized by the following features: bell-shaped male copulatory organ with a sleevelike base, accessory piece absent, vas deferens looping left intestinal cecum, vaginal aperture sinistroventral, and ventral anchor with elongate superficial and deep roots. PMID- 14533677 TI - Argentinochondria patagonensis n. gen., n. sp. (Copepoda: Chondracanthidae) parasitic on Genypterus brasiliensis (Pisces: Ophidiidae) from Patagonia, Argentina. AB - Argentinochondria patagonensis n. gen., n. sp. (Copepoda: Chondracanthidae) is described from adult specimens recovered from the smooth kingklip Genypterus brasiliensis Reagan, 1903, from San Jorge Gulf, Patagonia, Argentina. The new genus is placed in the Chondracanthinae. Females of the new genus differ from other members of this subfamily in having 2 pairs of modified legs, a bulbose head, a long neck region, and a trunk devoid of processes. Males have usual chondracanthid form, oral appendages as in female, except a longer paragnath, fewer teeth on maxilla, and 2 pairs of reduced legs. PMID- 14533678 TI - A new species of laelapine mite (Acari: Parasitiformes: Laelapidae) associated with Proechimys dimidiatus in the Atlantic forests of Brazil. AB - Tur megistoproctus, a new species of Laelapinae, is described from the pelage of the echimyid rodent Proechimys dimidiatus from the Atlantic forests of Ilha Grande, south of Rio de Janeiro. Measurements and illustrations are included for females and males. Another laelapine mite species, Tur turki Fonseca, co-occurred with T. megistoproctus in our studies and was recorded from the same host individuals and localities. These 2 laelapine mite species appear to be exclusively associated with a complex of echimyid rodent species (subgenus Trinomys) in the Atlantic forests of southeastern Brazil. PMID- 14533679 TI - Mucin-type O-glycosylation in helminth parasites from major taxonomic groups: evidence for widespread distribution of the Tn antigen (GalNAc-Ser/Thr) and identification of UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase activity. AB - This article focuses on the initiation pathway of mucin-type O-glycosylation in helminth parasites. The presence of the GalNAc-O-Ser/Thr structure, also known as Tn antigen, a truncated determinant related to aberrant glycosylation in mammal cells, and the activity of the UDP-GalNAc:polypeptide N-acetyl galactosaminyltransferase (ppGaNTase), the enzyme responsible for its synthesis, were studied in species from major taxonomic groups. Tn reactivity was determined in extracts from Taenia hydatigena, Mesocestoides corti, Fasciola hepatica, Nippostrongylus brasiliensis, and Toxocara canis using the monoclonal antibody 83D4. The Tn determinant was revealed in all preparations, and multiple patterns of Tn-bearing glycoproteins were observed by immunoblotting. Additionally, the first evidence that helminth parasites express ppGaNTase activity was obtained. This enzyme was studied in extracts from Echinococcus granulosus, F. hepatica, and T. canis by measuring the incorporation of UDP-(3H)GalNAc to both deglycosylated ovine syalomucin (dOSM) and synthetic peptide sequences derived from tandem repeats of human mucins. Whereas significant levels of ppGaNTase activity were detected in all the extracts when dOSM was used as a multisite acceptor, it was only observed in F. hepatica and E. granulosus extracts when mucin-derived peptides were used, suggesting that T. canis ppGaNTase enzyme(s) may represent a member of the gene family with a more restricted specificity for worm O-glycosylation motifs. The widespread expression of Tn antigen, capable of evoking both humoral and cellular immunity, strongly suggests that simple mucin type O-glycosylation does not constitute an aberrant phenomenon in helminth parasites. PMID- 14533680 TI - Prevalence of antibodies to Neospora caninum, Sarcocystis neurona, and Toxoplasma gondii in wild horses from central Wyoming. AB - Sarcocystis neurona, Neospora caninum, N. hughesi, and Toxoplasma gondii are 4 related coccidians considered to be associated with encephalomyelitis in horses. The source of infection for N. hughesi is unknown, whereas opossums, dogs, and cats are the definitive hosts for S. neurona, N. caninum, and T. gondii, respectively. Seroprevalence of these coccidians in 276 wild horses from central Wyoming outside the known range of the opossum (Didelphis virginiana) was determined. Antibodies to T. gondii were found only in 1 of 276 horses tested with the modified agglutination test using 1:25, 1:50, and 1:500 dilutions. Antibodies to N. caninum were found in 86 (31.1%) of the 276 horses tested with the Neospora agglutination test--the titers were 1:25 in 38 horses, 1:50 in 15, 1:100 in 9, 1:200 in 8, 1:400 in 4, 1:800 in 2, 1:1,600 in 2, 1:3,200 in 2, and 1:12,800 in 1. Antibodies to S. neurona were assessed with the serum immunoblot; of 276 horses tested, 18 had antibodies considered specific for S. neurona. Antibodies to S. neurona also were assessed with the S. neurona direct agglutination test (SAT). Thirty-nine of 265 horses tested had SAT antibodies--in titers of 1:50 in 26 horses and 1:100 in 13. The presence of S. neurona antibodies in horses in central Wyoming suggests that either there is cross reactivity between S. neurona and some other infection or a definitive host other than opossum is the source of infection. In a retrospective study, S. neurona antibodies were not found by immunoblot in the sera of 243 horses from western Canada outside the range of D. virginiana. PMID- 14533681 TI - Evidence of altered secondary lymphoid-tissue chemokine responsiveness in Balb/c mice infected with Schistosoma mansoni. AB - To determine the extent to which splenic T cells were affected by Schistosoma mansoni infection, we investigated the ability of the T cells to produce interferon (IFN)-gamma, as well as their chemotactic ability 7 wk PI. In this study, we report that splenic T cells from Balb/c mice with S. mansoni infections were capable of producing levels of IFN-gamma comparable with splenic T cells from naive mice. However, the T cells exhibited altered chemotactic activity, as evidenced by an inability to respond to secondary lymphoid-tissue chemokine (SLC/CCL21). Although no difference in chemokine expression was found between the spleens of infected versus control mice, chemokine production was greater in the livers of infected versus control mice. Collectively, these data indicate that Balb/c mice with 7-wk S. mansoni infection possess splenic T cells with altered chemotactic activity and that the alterations may be a consequence of the granulomatous response in the liver. PMID- 14533682 TI - In situ detection of antigenic glycoproteins in Taenia solium metacestodes. AB - Taenia solium has a complex life cycle. Its cysticercus can lodge in the brain, causing neurocysticercosis (NCC), and the adult tapeworm's survival in the intestine results in taeniasis. In this study, the in situ detection of previously described glycoprotein antigens used for serological diagnosis of NCC and the detection of other glycoconjugates was explored in cysticerci and the surrounding porcine tissue to understand their potential role in pathogenesis. Immunohistochemistry with an antiserum specific for glycoprotein antigens rich in N-linked carbohydrates and in situ histochemistry with a battery of lectins that have affinity to a variety of glycoconjugates were performed. The glycoconjugates rich in N-linked carbohydrates were detected in the vesicular fluid and tegument of the vesicular membrane and scolex, where the parasite has direct contact with the host tissues during cysticercosis and taeniasis, respectively. Additionally, as the inflammatory response progressed, the parasite's antigenic glycoproteins were also detected in the cytoplasm of inflammatory cells in the surrounding granuloma. In contrast, the spiral canal tegument, which will be exposed to intestinal enzymes in taeniasis, had N-acetyl-galactosamine-rich mucins. Thus, the differential saccharidic composition in T. solium metacestode structures may be important for the survival of the parasite in different host sites. PMID- 14533683 TI - Inhibition of Trypanosoma cruzi egress from infected fibroblasts is mediated by CD4+ and mu+ immune cells. AB - Trypanosoma cruzi, the causative agent of Chagas disease, is able to reproduce intracellularly in many host cell types while in the mammalian host. Although cellular immunity is known to be important in resistance to infection, the ability of immune cells to interfere with the completion of the intracellular growth cycle of T. cruzi has not been described. Using a tissue culture system to study the parasite growth cycle, we have found that spleen cells from infected mice are able to decrease the number of parasites released from infected fibroblasts. Spleen cells from mice infected for as few as 14 days and as long as 300 days display this inhibitory ability. Parasite egress from infected cells is inhibited by factor(s) released by immune cells during coculture with infected fibroblasts. Immune cell depletion studies indicate that the inhibitory activity requires the presence of both CD4+ T cells and mu+ B cells. These results suggest a direct ability of immune cells to somehow interfere with the completion of the intracellular cycle, and this ability may play a role in control of this parasite. PMID- 14533684 TI - Parasitological analysis of Leonese royalty from Collegiate-Basilica of St. Isidoro, Leon (Spain): helminths, protozoa, and mites. AB - The royal burial chamber of what is today the Collegiate-Basilica of St. Isidoro in Leon, Spain, built and remodeled between the 10th and 13th centuries and in the 20th century renamed the Kings' Pantheon, has 13 royal tombs that were opened in the presence of the Abbot-Prior of the Collegiate to enable a group of researchers to obtain all possible information from the royal remains. Several samples were sent to the Parasitology Unit of the Animal Pathology (Animal Health) Department at the Veterinary Faculty of Leon (Spain). In all the tombs, eggs and remains of nonparasitic mites were observed. In a piece of linen cloth from the bottom of 1 tomb, an Anoplocephala perfoliata egg was found. Furthermore, 4 mummified bodies were found. In 2 of these, those belonging to Infantes Maria and Fernando, Ascaris lumbricoides eggs were found and in the latter Trichuris trichiura eggs. We have not found in the literature reviewed any records of studies of this kind carried out in Spain. PMID- 14533685 TI - Tolerance to low temperatures of domestic and sylvatic Trichinella spp. in rat muscle tissue. AB - The present study was designed to investigate the tolerance to low temperatures of 9 Trichinella isolates in rat muscle tissue. Nine groups of 24 rats were infected with encapsulated Trichinella spiralis, Trichinella nativa, Trichinella britovi, Trichinella murrelli, Trichinella T6, Trichinella nelsoni, and 3 nonencapsulated Trichinella pseudospiralis strains. Six rats from each of the groups were necropsied at 5, 10, 20, and 40 wk postinfection (wpi). Muscle tissues containing Trichinella larvae were exposed to temperatures of -18, -5, and 5 C for 1 or 4 wk, and afterward the reproductive capacity index (RCI) in mice was determined for the 9 individual Trichinella isolates. Only T. nativa muscle larvae were infective after freezing at a temperature of -18 C. At 5 wpi all encapsulated isolates, except for the tropical species T. nelsoni, remained infective after exposure to a temperature of -5 C for both 1 and 4 wk, whereas nonencapsulated T. pseudospiralis survived only 1 wk of exposure. All Trichinella spp. remained infective after exposure to a temperature of 5 C. Muscle larvae for all investigated species remained infective as long as they persisted in live rats during the experiment. Analysis of variance showed a significant effect of age on the temperature tolerance of encapsulated T. spiralis and nonencapsulated T. pseudospiralis. In addition, significant interaction between age of muscle larvae and length of exposure was found. In general Trichinella muscle larvae of medium age (10 and 20 wpi) tolerated freezing better than early and late stages of infection (5 and 40 wpi). This is the first study to demonstrate such a relationship between age of infection and temperature tolerance of Trichinella spp. muscle larvae. PMID- 14533686 TI - Morphological description and life cycle of Paragonimus sp. (Trematoda: Troglotrematidae): causal agent of human paragonimiasis in Colombia. AB - The first morphological description is made of all stages of the life cycle of a Paragonimus species infecting humans in Colombia. Larval stages were obtained both in vitro and from field collections. Adult Paragonimus spp. are described. The aquatic snail Aroapyrgus sp. serves as an intermediate host of this species, both naturally and experimentally, yielding rediae and cercariae. Crabs (Hypolobocera bouvieri monticola and H. emberarum) were found to be the natural second intermediate hosts, and individuals of another crab species (Strengeria sp.) were also infected in the laboratory. PMID- 14533687 TI - Seasonal and yearly population dynamics of two exotic helminths, Camallanus coti (Nematoda) and Bothriocephalus acheilognathi (Cestoda), parasitizing exotic fishes in Waianu Stream, O'ahu, Hawaii. AB - Exotic poeciliid fishes introduced into Hawaiian freshwaters are responsible for the introduction of several exotic parasites, of which the most important are Camallanus cotti and Bothriocephalus acheilognathi in terms of potential disease threat to native stream fishes. This roundworm and tapeworm are the most prevalent and abundant freshwater fish helminths in Hawaiian streams. This study examined the seasonal and yearly population structure of C. cotti and B. acheilognathi to determine if the tropical Hawaiian environment characterized by low climatic variability permits continuous opportunities for parasite transmission regardless of time of year. Camallanus cotti displayed seasonal differences in prevalence and mean abundance, whereas B. acheilognathi did not. Camallanus cotti prevalence and mean abundance were higher in the Hawaiian summer (47.7%, 0.79) than in winter (25.8%, 0.36). A seasonal relationship of C. cotti levels is likely explained by extensive rains associated with the Hawaiian winter season, which may act to decrease parasite transmission by flushing infected poeciliid hosts, intermediate copepod hosts, and possibly free-living infective worm stages downstream. Bothriocephalus acheilognathi displayed low prevalence and mean abundance in both summer (4.0%, 0.06) and winter (6.5%, 0.07), and it may be difficult to detect seasonal changes due to these low levels. Camallanus cotti prevalence and mean abundance remained relatively constant from the summer of 1995 to the summer of 1999, indicating that levels of this roundworm are stable in Waianu Stream. Whereas B. acheilognathi prevalence and mean abundance were low during the summer of 1995 and the summer of 1997, a dramatic peak in prevalence and mean abundance was observed in the summer of 1998 (41.2%, 1.06), with levels decreasing sharply in the summer of 1999 (4.4%, 0.07). It appears that B. acheilognathi also is present in stable populations at low levels, even though levels rose sharply during a single year. PMID- 14533688 TI - Differential gene expression during desiccation stress in the insect-killing nematode Steinernema feltiae IS-6. AB - The developmentally arrested life stage of the entomopathogenic nematode Steinernema feltiae is exposed to threats of survival, including desiccation. We adopted a comprehensive approach to the study of the molecular mechanisms of desiccation stress tolerance in S. feltiae IS-6. We identified, expressed sequence tags (ESTs) that are differentially expressed during desiccation stress in S. feltiae IS-6 infective juveniles using DNA subtractive hybridization. These ESTs included genes that are known to be stress related, genes that are homologous to hypothetical Caenorhabditis elegans proteins, and novel genes that may be involved in traits specific to S. feltiae. Expression pattern characterization revealed that all analyzed ESTs were induced during 8 and 24 hr of dehydration of S. feltiae IS-6. Our results unveiled some of the components of the genetic networks that are activated in S. feltiae IS-6 during dehydration and suggested a differing pattern of temporal regulation during nematode dehydration. PMID- 14533689 TI - Cis and trans factors involved in apicoplast targeting in Toxoplasma gondii. AB - Ribosomal subunit protein 9 (rps9) is a nuclearly encoded protein that resides in the apicoplast organelle of Toxoplasma gondii. Two cis-acting regions within the rps9 transit domain (amino acids 38-49 and 79-86), when combined with the rps9 signal sequence, were necessary and sufficient for apicoplast targeting. To investigate proteins interacting with the rps9 leader sequence, parasites expressing rps9 leader constructs fused to a glutathione S-transferase (GST) reporter were prepared, and proteins associated with the leader constructs were purified from extracts by affinity chromatography. In addition to GST-containing peptides, proteins with apparent masses of 92, 90, 86, and 160 kDa were purified. Mass spectrometry data suggested that the 92- and 90-kDa polypeptides appear to be subtilisin-like proteins, whereas the 86-kDa polypeptide was identified as the molecular chaperone BiP of T. gondii. PMID- 14533690 TI - Low virulence of oocysts of Czech Toxoplasma gondii isolates on the basis of biological and genetic characteristics. AB - The virulence of the oocysts of 7 Czech Toxoplasma gondii isolates was tested. The oocysts were obtained by experimental infection of cats with the tissue cysts of T. gondii isolates from dogs, cats, and rabbits. The cats shed the oocysts in feces, with prepatent periods of 3-5 days postinfection (PI); the patent period was 7-18 days. The number of oocysts shed varied between 0.94 million and 47 million, with 0.66 million-39 million oocysts found in the daily samples of excrement. The cats ceased oocyst production at 11-22 days PI. Sporulated oocysts were used to prepare infective doses of 1 to 10(5) oocysts for oral infection of 10 mice. Deoxyribonucleic acid isolated from 4 T. gondii isolates was used in polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for amplification of the ROP1 gene and restriction of the product of amplification by restriction endonuclease DdeI. On the basis of their biological characteristics, all 7 isolates belonged to the group of "avirulent" strains. In the PCR-RFLP tests, 2 isolates, K9 and K19, showed an "avirulent" strain pattern. PMID- 14533691 TI - Spinitectus macrospinosus n. sp. (Nematoda: Cystidicolidae) from the channel catfish Ictalurus punctatus in southern Manitoba and its distribution in other Ictalurus spp. AB - Spinitectus macrospinosus n. sp., a parasite of the channel catfish Ictalurus punctatus, from the Red and Assiniboine rivers in southern Manitoba, Canada, is characterized by spines reaching a maximum length of 21.8-26.8 microm (means for males and females, respectively) in the anterior esophageal region, arranged in 4 sectors in the anterior region of the body. Spines in this region of the body increase in size and decrease in number (from a maximum of 6 to 3 per sector). The combination of spine length, number, and arrangement and the position of the excretory pore (between spine rows 6 and 7) readily distinguishes it from all its North American congeners, including the species it is most similar to, S. carolini. It is different from S. gracilis and S. acipenseri in having a posteriorly directed vagina, an excretory pore between spine rows 6 and 7, a longer stoma, a right spicule with a terminal ventral barb, and a heart-shaped caudal mucron. Spinitectus macrospinosus is similar to other North American species of Spinitectus, e.g., S. carolini, S. micracanthus, S. mexicanus, S. osorioi, and S. humbertoi, in having the anterior rows of spines arranged in 4 sectors. It is also similar to S. micracanthus and S. carolini in possessing a relatively long stoma, a posteriorly directed vagina, a right spicule with a terminal ventral barb, and a heart-shaped caudal mucron. Furthermore, it is similar to S. carolini in possessing an "area rugosa" with 2 rows of precloacal cuticular cleats. In southern Manitoba, S. macrospinosus appears to mature only in the channel catfish. Reexamination of museum specimens revealed that the nematode also is found in I. furcatus in Kentucky Lake (Kentucky-Tennessee) and in I. lacustris in Lake Texoma (Oklahoma-Texas). PMID- 14533692 TI - The taxonomic status of Digramma (Pseudophyllidea: Ligulidae) inferred from DNA sequences. AB - The traditional classification of the ligulid tapeworms into 2 genera, Ligula Bloch, 1782 and Digramma Cholodkovsky, 1914, remains controversial. Molecular data of sequences for the 5' end of the nuclear 28S ribosomal ribonucleic acid (rRNA) gene, the mitochondrial cytochrome c oxidase subunit I (COI) gene, and the nicotinamide adenine dinucleotide dehydrogenase subunit 1 (ND1) gene, as well as the first internal transcribed spacer (ITS1) of the nuclear ribosomal deoxyribonucleic acid (DNA), were used to characterize Digramma and to investigate its relationship with Ligula. Digramma spp. exhibited identical sequences with Ligula intestinalis both in the 28S rRNA and the COI gene and differed from L. intestinalis by 0.7% in the ITS1 region and 7.4% in the ND1 gene, respectively. A high degree of genetic conservation within 28S ribosomal DNA, COI, ITS1, and even ND1 genes, was found in Ligula and Digramma. The low genetic divergence in the 4 genes between Ligula and Digramma indicates that Digramma is probably not an independent genus. Therefore, it is proposed that Ligula and Digramma should be considered as 2 species within the genus Ligula and the tapeworms of Digramma collected from diverse localities in China belong to the same species. The present study also suggests that ITS1 and ND1 sequences can act as useful genetic markers to distinguish Ligula and Digramma. PMID- 14533693 TI - Echinostoma luisreyi n. sp. (Platyhelminthes: Digenea) by light and scanning electron microscopy. AB - The entire life cycle of the Brazilian Echinostoma luisreyi n. sp., of the 37 collar spine E. revolutum group, has been observed under experimental conditions. The snail Physa marmorata serves as its first natural intermediate host. This species and the planorbid Biomplalaria glabrata act as experimental second intermediate hosts. The worm recovery rate was 51.3 and 0.6% for mice and hamsters, respectively, but the infection did not develop in the quail Coturnix coturnix. The natural vertebrate host is not known. The most important morphological character of the new species separating it from the other Echinostoma species studied is the oral corner spines that increase in size from the latero-oral to the ventro-oral regions. The ratio of the sizes of the smaller oral spines and the larger aboral ones was 1.7:1.0. Also, the excretory pore is radially wrinkled and dorsally subterminal. Images obtained by scanning electron microscopy confirmed the characters that differentiate the new species and the most closely related species. PMID- 14533694 TI - A redescription of Cryptosporidium galli Pavlasek, 1999 (Apicomplexa: Cryptosporidiidae) from birds. AB - Cryptosporidium galli Pavlasek, 1999, described from the feces of birds, is redescribed with additional molecular and biological data. Oocysts are ellipsoidal, are passed fully sporulated, lack sporocysts, and measure 8.25 x 6.3 microm (range 8.0-8.5 x 6.2-6.4 microm) with a length-width ratio of 1.30 (n = 50). Oocysts are structurally similar to those of Cryptosporidium baileyi described from chickens, but in addition to being considerably larger than oocysts of C. baileyi, these oocysts infect the proventriculus in a variety of birds and not the respiratory tract. Oocysts were successfully transmitted from chickens to chickens, and morphologically similar oocysts also were observed in a variety of exotic and wild birds (Order Passeriformes, Phasianidae, Fringillidae, and Icteridae). Molecular and phylogenetic analyses at the 18S rRNA, HSP70, and actin gene loci demonstrate that this species is genetically distinct from all known species and genotypes of Cryptosporidium and, thus, was named C. galli. PMID- 14533695 TI - Whallwachsia illuminata n. gen., n. sp. (Trematoda: Digenea: Plagiorchiformes: Prosthogonimidae) in the steely-vented hummingbird Amazilia saucerrottei (Aves: Apodiformes: Trochilidae) and the yellow-olive flycatcher Tolmomyias sulphurescens (Aves: Passeriformes: Tyraninidae) from the Area de Conservacion Guanacaste, Guanacaste, Costa Rica. AB - A new species of digenean found in the intestines of the steely-vented hummingbird Amazilia saucerrottei and the yellow-olive flycatcher Tolmomyias sulphurescens from the Area de Conservacion Guanacaste, Guanacaste, Costa Rica, resembles members of the Prosthogonimidae in having a highly lobate ovary; an elongate cirrus sac containing the cirrus, pars prostatica, and internal seminal vesicle; no external seminal vesicle; 2 fields of extracecal vitelline follicles restricted to the area between the intestinal bifurcation and testes; and uterine loops occupying all available space in the hind body. The new species differs from all other members of the family in having genital pores opening laterally to the cecum, immediately anterior to the acetabular level, and markedly oblique rather than symmetrical testes. Consequently, we propose the new genus Whallwachsia for the species. Preliminary phylogenetic assessment suggests that the species is the sister group of all other prosthogonimids. PMID- 14533696 TI - Tinamutrema canoae n. gen. et n. sp. (Trematoda: Digenea: Strigeiformes: Brachylaimidae) in Crypturellus cinnamomeus (Aves, Passeriformes, Tinamidae) from the Area de Conservacion Guanacaste, Costa Rica. AB - We propose Tinamutrema as a new genus for Brachylaima centrodes (Braun, 1901) Dollfus, 1935 and for T. canoae, as a new species inhabiting tinamus in the Area de Conservacion Guanacaste, Costa Rica. Specimens from Costa Rica resemble B. centrodes in having an elongate body, pretesticular genital pore and terminal genitalia, intercecal uterine loops occupying all available space between the anterior testis and the intestinal bifurcation, an oral sucker width:pharynx width ratio of approximately 1:0.55, an oral sucker:ventral sucker width ratio of approximately 1:1, and vitelline follicles extending into the forebody closer to the pharynx than to the anterior margin of the ventral sucker and by living in the cloaca. They differ from B. centrodes in having vitelline follicles that do not extend as far anteriorly as those in B. centrodes, which extend anteriorly to the level of the anteriormost extent of the cecal "shoulders," dense tegumental spination as opposed to sparse or no spination, relatively smaller cirrus with fewer spines, longer and more sinous pars prostatica, and forebody averaging 36% of total body length (TBL) as opposed to 42% TBL. Both species differ from other members of the Brachylaimidae in possessing a spinose cirrus and a cirrus sac containing both the cirrus and the pars prostatica. Preliminary phylogenetic assessment suggests that these traits are plesiomorphic, and thus the species are basal to the rest of the Brachylaimidae, whose diagnosis we emend accordingly. PMID- 14533697 TI - Bakkeius moragai n. gen. et n. sp. and Pojmanskia riosae n. gen. et n. sp. (Trematoda: Digenea: Brachylaimoidea) in birds from the Area de Conservacion Guanacaste, Costa Rica. AB - We describe 2 new species of leucochloridiid-like brachylaimoid digeneans parasitizing a variety of birds in the Area de Conservacion Guanacaste, Costa Rica, each of which we assign to a new genus. According to Pojmanska's (Pojmanska, T. 2002a. Superfamily Brachylaimoidea Joyeux & Foley, 1930. In Keys to the Trematoda, D. I. Gibson, A. Jones, and R. A. Bray [eds.]. CAB International and The Natural History Museum, London, U.K., p. 31-36.) key for the Brachylaimoidea, we are unable to place either species in any family. One species most closely resembles members of Leucochloridium by having well developed suckers, lacking an esophagus, and having cecal shoulders, gonads at the posterior end, and the genital pore at posterior end of body but differs by having symmetrical testes, a posttesticular ovary, and a terminal genital pore; thus, we propose the genus Bakkeius for it. The second new genus resembles members of Michajlovia by having ventral genital pores but differs by having extracecal uterine loops in the forebody, a cirrus sac containing the pars prostatica and seminal vesicle, and gland cells surrounding the genital pore; thus, we propose Pojmanskia for it. These new genera must currently be treated as incertae sedis according to Pojmanska (op. cit.); however, we feel that future phylogenetic analyses will require emendation of the family diagnosis for Leucochloridiidae to include those taxa with terminal and ventral genital pores and with preovarian testes. PMID- 14533698 TI - Neohaematotrephus arayae n. sp. (Digenea: Echinostomiformes: Cyclocoelidae) in Jacana spinosa (Aves: Charadriiformes: Jacanidae) from the Area de Conservacion Guanacaste, Costa Rica. AB - Specimens of a species of cyclocoelid digenean inhabiting Jacana spinosa from the Area de Conservacion Guanacaste, Costa Rica, most closely resemble Haematotrephus facioi (Brenes and Arroyo, 1962) Yamaguti, 1971, in the same host from Aranjuez, Puntarenas Province, Costa Rica, in having confluent vitelline follicles posteriorly, diagnostic of Neohaematotrophus, and in pharynx length, ovary width, and cirrus sac on the sinistral side. The new species is also highly similar in appearance to H. gendrei Dubois, 1959, also inhabiting a jacanid (from West Africa), which has vitelline follicles confluent posteriorly, and extending anteriorly to the intestinal bifurcation and genital pore opening immediately posterior to the anterior margin of the pharynx. Like H. facioi, H. gendrei has a relatively much shorter and broader cirrus sac than does the new species. Examination of the holotype and paratype of H. facioi confirmed that the specimens from Guanacaste differ in having a longer body, a larger ovary and eggs, and smaller testes. They also have the ovary on the sinistral rather than the dextral side of the body, genital pore anterior to the pharynx rather than at or posterior to the level of the posterior margin of the pharynx, longer and thinner cirrus sac, and eggs without eyespotted miracidia. Half the eggs in both specimens of H. facioi have well-developed eyespotted miracidia, whereas the typical condition for cyclocoelids is for virtually all eggs to exhibit eyespotted miracidia. Both H. facioi and H. gendrei are transferred to Neohaematotrophus, along with the new species. PMID- 14533699 TI - The use of immunoblot analysis in the diagnosis of canine visceral leishmaniasis in an endemic area of Rio de Janeiro. AB - In this article, we describe the findings obtained using immunoblot analysis in the diagnosis of canine visceral leishmaniasis (VL) and its correlation with serological titer and clinical status. We found that all animals bearing amastigote forms recognized antigens with 29 and 32 kDa and that this pattern can be exploited for diagnostic and epidemiological purposes. The recognition of the 29- and 32-kDa antigens was verified even in seronegative dogs and preceded seroconversion in periods ranging from several months to 2 yr. We found a correlation between serological titer and parasite burden. Although no correlation between antigenic recognition pattern and clinical status was observed, immunoblot analysis proved to be a reliable test to detect antibodies against Leishmania sp. antigens in dogs from areas with endemic VL. PMID- 14533701 TI - Optimized serodiagnosis of sheep fascioliasis by Fast-D protein liquid chromatography fractionation of Fasciola hepatica excretory-secretory antigens. AB - Current methods for the serodiagnosis of sheep fascioliasis show suboptimal sensitivity, specificity, or both. With the aim of developing an improved method, we fractionated native Fasciola hepatica excretory-secretory antigens (ESAs) by size-exclusion FPLC (fast protein liquid chromatography) on a Superdex 75 HR 10/30 column and then tested the serodiagnostic value of the antigens contained in each one of the 4 peaks obtained (peaks I-IV). Serodiagnostic value was assessed using sera from sheep naturally infected with F. hepatica (group A); sera from the individuals of a fluke-free herd (most of which also had other intestinal nematodes, lung nematodes, Moniezia spp., and/or Cysticercus tenuicollis) sera from a fluke-free herd (group B); sera from lambs experimentally infected with 10-40 F. hepatica metacercariae (group C); and sera from uninfected control lambs (group D). Enzyme-linked immunosorbent assay (ELISA) with peak I or II as target antigens (and to a lesser extent with peak III as target) showed reactivity with negative sera, so that it was not possible to establish cutoff values discriminating infected and uninfected animals. In contrast, when peak IV was used as target, a low cutoff value of 0.235 optical density units (mean + 4 SD) discriminated infected and uninfected animals, with 100% sensitivity and 100% specificity. ELISA with peak IV as a target identified infected animals (even animals that had received only 10 metacercariae) within 3 5 wk of infection and subsequently throughout the rest of the 14-wk monitoring period. In Western blotting analysis, again only the antigens contained in peak IV (range 7-40 kDa, under reducing conditions) were specific for diagnosis of infected animals. These results indicate that molecular sieving of F. hepatica ESAs by this procedure is a fast, simple, reproducible way of obtaining antigens useful for serodiagnosis of sheep fascioliasis. PMID- 14533700 TI - Activities of therapeutic agents against Naegleria fowleri in vitro and in a mouse model of primary amebic meningoencephalitis. AB - Inhalation of water contaminated with Naegleria fowleri may lead to a potentially fatal infection of the central nervous system known as primary amebic meningoencephalitis (PAM). Amphotericin B (AMB), an antifungal drug, is the only agent with established clinical efficacy in the treatment of PAM, though therapy with this drug is not always effective and has been associated with adverse effects on the kidneys and other organs. We investigated the activity of various therapeutic agents against N. fowleri in an attempt to identify other useful agents for treating PAM. Several of these agents exhibited in vitro activity against the Lee (M67) strain of N. fowleri. The minimum inhibitory concentrations of these agents were 0.1 microg/ml (ketoconazole), 1 microg/ml (liposomal AMB), and 10 microg/ml (minocycline, quinupristin-dalfopristin, and trifluoperazine). Other agents had a minimum inhibitory concentration > 10 microg/ml (linezolid) or > 100 microg/ml (rifampin). In a mouse model of PAM, none of the untreated control mice survived, whereas the survival of treated animals was 50% (quinupristin-dalfopristin), 30% (ketoconazole and liposomal AMB), 20% (trifluoperazine), and 10% (linezolid and minocycline). Further studies are needed to ascertain whether these agents have synergistic activity with AMB in vitro and in vivo. PMID- 14533702 TI - Seroprevalence of Toxoplasma gondii antibodies in the rodent capybara (Hidrochoeris hidrochoeris) from Brazil. AB - Capybaras (Hidrochoeris hidrochoeris) are 1 of the largest rodents used for meat in South and Central America. Prevalence of anti-Toxoplasma gondii antibodies in 149 feral H. hidrochoeris from the state of Sao Paulo, Brazil, was evaluated using the indirect immunofluorescent antibody test (IFAT) and the modified agglutination test (MAT). Using IFAT, antibodies (>1:16) were found in 104 (69.8%) and with the MAT, antibodies (>1:25) were found in 63 (42.3%) capybaras. This is the first report of prevalence of T. gondii antibodies in this host. PMID- 14533703 TI - Toxoplasma gondii isolates of free-ranging chickens from Rio de Janeiro, Brazil: mouse mortality, genotype, and oocyst shedding by cats. AB - Most isolates of Toxoplasma gondii can be grouped into 3 genetic lineages. In the present study, 67 isolates of T. gondii were obtained by bioassay in mice inoculated with brains and hearts of 96 asymptomatic chickens from an area highly endemic to human infection in Rio de Janeiro, Brazil. Of the 48 isolates genotyped using the SAG2 locus, 34 (70%) were of type I and 13 (27%) were of type III. No isolate of type II was recovered. Isolates from 1 chicken contained a type I and type III mixed infection, indicating natural multiparasite infection in the same animal. Cats fed mice infected with 11 type I strains shed 19-535 million oocysts in their feces, indicating that type I isolates can circulate in the environment. PMID- 14533704 TI - Effect of vaccination with a recombinant fusion protein encoding an astacinlike metalloprotease (MTP-1) secreted by host-stimulated Ancylostoma caninum third stage infective larvae. AB - Laboratory dogs were vaccinated intramuscularly with a recombinant fusion protein (expressed and isolated from Escherichia coli) formulated with the Glaxo SmithKline Adjuvant System 02 (AS02). The fusion protein encoded Ac-MTP-1, a developmentally regulated astacinlike metalloprotease secreted by host-stimulated Ancylostoma caninum third-stage larvae (L3). Control dogs were injected intramuscularly with an equivalent amount of AS02 adjuvant alone. The vaccinated and control dogs were then challenged by s.c. injection of 500 L3 of the canine hookworm A. caninum. The vaccinated dogs developed prechallenge immunoglobulin G2 (IgG2) antibody responses specific to anti-Ac-MTP-1-fusion protein with titers ranging between 1:40,000 and 1:364,000, whereas they developed antigen-specific immunoglobulin E antibody responses with titers ranging between 1:500 and 1:1,500. By immunoblotting, canine sera obtained from the vaccinated dogs recognized a protein of the estimated apparent molecular weight of Ac-MTP-1 in activated L3 secretory products. Spearman rank order correlations between the canine intestinal adult hookworm burden and quantitative egg counts at necropsy and anti-Ac-MTP-1 IgG2 antibody titers revealed a statistically significant inverse association (r = -0.89; P = 0.04), suggesting that this molecule offers promise as a recombinant vaccine. PMID- 14533705 TI - Neurotoxicity and immunotoxicity assessment in CBA/J mice with chronic Toxoplasma gondii infection and multiple oral exposures to methylmercury. AB - The present study was conducted to determine the effect of multiple low doses of methylmercury (MeHg) on the course of a chronic Toxoplasma gondii infection. Four groups of 6-wk-old female CBA/J mice either were fed 25 T. gondii tissue cysts of the ME-49 strain or were vehicle control. Six weeks later, half of each group was orally gavaged with 8-mg/kg body weight doses of MeHg on days 0, 2, 4, 7, 10, and 13, totaling 4 experimental groups. Mice were killed on day 17 or 18 after MeHg exposure. Flow cytometric analysis of lymphocyte subpopulations in the thymus demonstrated a significant increase in the percentage of CD4- CD8+ T-cells in mice exposed to MeHg with a concurrent T. gondii infection. Groups of mice exposed to MeHg showed a decrease in total thymic cellularity and cellularity of all T-cell subpopulations when compared with control mice, but viability of these cells was unaffected. Splenic cell viability was decreased in mice exposed to MeHg, but alterations in T-cell subpopulations were not noted. These data indicate that multiple low doses of MeHg may not exacerbate chronic toxoplasmosis, but MeHg-induced effects on the immune system were evident. PMID- 14533706 TI - Meningoencephalitis associated with an unidentified apicomplexan protozoan in a Pacific harbor seal. AB - A Pacific harbor seal (Phoca vitulina richardsii) was found on the central California coast with neurologic signs and labored breathing, which were unresponsive to treatment. Necropsy revealed a nonsuppurative necrotizing meningoencephalitis, a multilocular thymic cyst, and nonsuppurative cystitis and renal pyelitis. Microscopic examination revealed protozoans in the brain, thymic cyst, and bladder mucosa. Ultrastructurally, the protozoal tachyzoites were different from those of Neospora caninum, Toxoplasma gondii, and Sarcocystis neurona; the rhoptries were small and had electron-dense contents, and the organism divided by endodyogeny. Specific antibodies were not detected in serum using agglutination (N. caninum, T. gondii) and immunoblot assays (S. neurona). Immunohistochemistry for these organisms was negative. Polymerase chain reaction on brain tissue using specific primers did not amplify T. gondii deoxyribonucleic acid. The meningoencephalitis in this seal thus appears to have been caused by a novel protozoan. PMID- 14533707 TI - A gastropod scavenger serving as paratenic host for larval helminth communities in shore crabs. AB - The whelk Cominella glandiformis is an important predator-scavenger of New Zealand intertidal ecosystems; a few whelks can quickly eat all the soft tissues of recently dead crabs. In this study, we demonstrate that whelks can also ingest and act as paratenic hosts for at least 4 helminth species that use crabs as intermediate hosts: metacercariae of the trematode Maritrema sp. and of another unidentified trematode, larval acuariid nematodes, and cystacanths of the acanthocephalans Profilicollis spp. Large whelks ingest disproportionately more helminth larvae than small whelks, but the survival of parasites during their short stay in the whelks is not affected by whelk size. The majority of metacercariae and nematodes are passed out in whelk feces within 3 days of ingestion, whereas the few cystacanths found did not leave whelks until after that time; no parasite was left in whelks 5 days postingestion. Survival of all 4 helminth species was generally very high, though it decreased day by day in 2 species. Given that the avian definitive hosts of all 4 helminths also eat whelks, our results indicate that alternative transmission pathways exist and that parasites can take routes through food webs that are too often ignored. PMID- 14533708 TI - Autofluorescence of Toxoplasma gondii and related coccidian oocysts. AB - This is the first report of blue autofluorescence as a useful characteristic in the microscopic detection of Toxoplasma gondii, Hammondia hammondi, Hammondia heydorni, Neospora caninum, Besnoitia darlingi, and Sarcocystis neurona oocysts or sporocysts. This autofluorescence is of sufficient intensity and duration to allow identification of these oocysts from complex microscopic sample backgrounds. As with the autofluorescence of related coccidia, the oocysts glow pale blue when illuminated with an ultraviolet (UV) light source and viewed with the correct UV excitation and emission filter set. PMID- 14533709 TI - Ascorbic acid is a requirement for the morphogenesis of the human filarial parasite Brugia malayi. AB - The nematode parasites Wuchereria bancrofti, Brugia malayi, and B. timori cause a disease in humans known as lymphatic filariasis, which afflicts approximately 120 million people worldwide. The parasites enter the human host from the mosquito either as L3 or as infective larvae and subsequently differentiate through 2 molts. In this article, we show that B. malayi depends on an exogenous source of vitamin C to complete the L3 to L4 molt, a critical morphogenic step in its life cycle. Brugia malayi apparently belongs to a small group of living organisms that depend on an exogenous source of vitamin C. This group includes only primates (including man) and guinea pigs among mammals. PMID- 14533710 TI - Seroprevalence of Neospora caninum and Toxoplasma gondii antibodies in the South American opossum (Didelphis marsupialis) from the city of Sao Paulo, Brazil. AB - Antibodies to Neospora caninum and Toxoplasma gondii were assayed in sera of 396 opossums (Didelphis marsupialis) from the city of Sao Paulo, Brazil. Antibodies to N. caninum were assayed using the indirect immunofluorescent antibody test (IFAT). Antibodies (IFAT, approximately 1:25) to N. caninum were found in 84 opossums (D. marsupialis) in titers of 1:25 in 46, 1:50 in 20, 1:100 in 17, and 1:400 in 1. Antibodies to T. gondii were assayed with the modified agglutination test (MAT) and the IFAT. Antibodies to T. gondii (MAT, approximately 1:25) were found in 82 (20.4%) of the 396 opossums, in titers of 1:25 in 24, 1:50 in 26, 1:100 in 18, 1:200 in 13, and 1:800 in 1. The IFAT antibodies to T. gondii were found in 148 of 396 opossums, in titers of 1:16 in 41, 1:32 in 23, 1:64 in 13, 1:128 in 6, 1:256 in 20, 1:512 in 17, 1:1,024 in 10, 1:2,048 in 10, 1:4,096 in 7, and 1:8,192 in 1. This is the first report of N. caninum and T. gondii infections in D. marsupialis. PMID- 14533711 TI - Chromosomal polymorphism in the yeast species Debaryomyces hansenii. AB - Pulse field gel electrophoresis karyotypes of 41 strains of the genus Debaryomyces, including 35 strains confirmed as D. hansenii species by D1/D2 ribosomal DNA sequence analysis, were performed. Electrophoretic karyotypes of the 41 strains exhibited 4 to 10 chromosomal bands ranging between 0.7 Mb and 4.2 Mb. Among D. hansenii species, the patterns of strains obtained from the CBS collection and cheese isolates differed strongly from D. hansenii var. hansenii CBS767T. Both D. hansenii var. hansenii and D. hansenii var. fabryii showed chromosome length polymorphism. Electrophoretic karyotypes of the D. hansenii strains were analyzed by Southern hybridization with various species-specific probes isolated from D. hansenii var. hansenii CBS767T. Repeated sequences including the F01pro, M18pro, the Ty1-copia retrotransposon Tdh5 and hypothetical telomeric sequence hybridized to several chromosomal bands, while a D1/D2 probe derived from the large ribosomal sub-unit hybridized only to the largest chromosome. Unique probes such as those hybridizing to actin ACT1, glycerol-3 phosphate dehydrogenase GPD1 and beta-glucosidase LAC4 encoding genes were assigned to specific chromosomal bands of D. hansenii var. hansenii CBS767T. These probes failed to hybridize to D. hansenii var. fabryii strongly suggesting that strains of this variety actually represent a different taxon. PMID- 14533712 TI - Nitrile hydrolysing activities of deep-sea and terrestrial mycolate actinomycetes. AB - Nitrile metabolising actinomycetes previously recovered from deep-sea sediments and terrestrial soils were investigated for their nitrile transforming properties. Metabolic profiling and activity assays confirmed that all strains catalysed the hydrolysis of nitriles by a nitrile hydratase/amidase system. Acetonitrile and benzonitrile, when used as growth substrates for enzyme induction experiments, had a significant influence on the biotransformation activities towards various nitriles and amides. The specific activities of selected deep-sea and terrestrial acetonitrile-grown bacteria against a suite of nitriles and amides were higher than those of the only other reported marine nitrile-hydrolysing R. erythropolis, isolated from a shallow sediment. The increase of nitrile chain length appeared to have negative influence on the nitrile hydratase activity of acetonitrile-grown bacteria, but the same was not true for benzonitrile-grown bacteria. The nitrile hydratases and amidases were constitutive in 10 of the 16 deep-sea and terrestrial actinomycetes studied, and one strain showed an inducible hydratase and a constitutive amidase. Most of the deep-sea strains had constitutive activities and showed some of the highest activities and broadest substrate specificities of organisms included in this study. PMID- 14533713 TI - Differential pulse polarography: a method for the direct study of biosorption of metal ions by live bacteria from mixed metal solutions. AB - The technique of differential pulse polarography is shown here to be applicable to the monitoring directly the biosorption of metal ions from solution by live bacteria from mixed metal solutions. Biosorption of Cd(II), Zn(II) and Ni(II) by P. cepacia was followed using data obtained at the potential which is characteristic of the metal ion in the absence and presence of cells. Hepes buffer (pH 7.4, 50 mM) was used as a supporting electrolyte in the polarographic chamber and metal ion peaks in the presence of cells of lower amplitude were obtained due to metal-binding by the cells. Well defined polarographic peaks were obtained in experiments involving mixtures of metal ions of Cd(II)-Zn(II), Cu(II) Zn(II), Cu(II)-Cd(II) and Cd(II)-Ni(II). Biosorption of Cd(II), Zn(II) increased with solution pH. The method was also tested as a rapid technique for assessing removal of metal ions by live bacteria and the ability of the polarographic technique in measuring biosorption of metal ions from mixed metal solutions is demonstrated. Cu(II) was preferentially bound and removal of metals was in the order Cu(II) > Ni(II) > Zn(II), Cd(II) by intact cells of P. cepacia. PMID- 14533714 TI - Mycogloea nipponica--the first known teleomorph in the heterobasidiomycetous yeast genus Kurtzmanomyces. AB - A culture was obtained from a spore print of a basidiocarp of Mycogloea nipponica collected in Taiwan. A yeast stage and basidia identical to those of M. nipponica developed in laboratory media. The Taiwanese specimen of M. nipponica and its yeast anamorph were characterised in the present study. Comparative morphological, molecular, and ultrastructural studies indicated that the yeast stage can be assigned to the genus Kurtzmanomyces. The revealed connection between the sexual species Mycogloea nipponica and the asexual genus Kurtzmanomyces demonstrates the importance of anamorphic characteristics in the modem systematics of heterobasidiomycetous fungi. PMID- 14533715 TI - Low sequence similarity and gene content of symbiotic clusters of Bradyrhizobium sp. WM9 (Lupinus) indicate early divergence of "lupin" lineage in the genus Bradyrhizobium. AB - Two sequenced nodulation regions of lupin Bradyrhizobium sp. WM9 carried the majority of genes involved in the Nod factor production. The nod region I harbored: nolA, nodD, nodA, nodB, nodC, nodS, nodI, nodJ, nolO, nodZ, fixR, nifA, fixA, nodM, nolK and noeL. This gene arrangement resembled that found in the nodulation region of Bradyrhizobium japonicum USDA110, however strain WM9 harbored only one nodD gene copy, while the nodM, nolK and noeL genes had no counterparts in the 410 kb symbiotic region of strain USDA110. Region II harbored nolL and nodW, but lacked an nodV gene. Both regions carried ORFs that lacked similarity to the published USDA110 sequences, though they had homologues in symbiotic regions of Rhizobium etli, Sinorhizobium sp. NGR234 and Mesorhizobium loti. These differences in gene content, as well as a low average sequence identity (70%) of symbiotic genes with respect to B. japonicum USDA110 were in contrast with the phylogenetic relationship of USDA110 and WM9 revealed by the analysis of 16S rDNA and dnaK sequences. This most likely reflected an early divergence of symbiotic loci, and possible co-speciation with distinct legumes. During this process the loss of a noel gene and the acquisition of a nolL gene could be regarded as an adaptation towards these legumes that responded to Nod factors carrying 4-O-acetylfucose rather than 2-O-methylfucose. This explained various responses of lupins and serradella plants to infection by mutants in nodZ and nolL genes, knowing that serradella is a stringent legume while lupins are more promiscuous legumes. PMID- 14533716 TI - Osmotolerance and leavening ability in sweet and frozen sweet dough. Comparative analysis between Torulaspora delbrueckii and Saccharomyces cerevisiae baker's yeast strains. AB - The response of Saccharomyces cerevisiae and freeze-tolerant Torulaspora delbrueckii strains to osmotic stress and their CO2 production capacity in sweet and frozen-sweet dough has been examined. T. delbrueckii strains, IGC5321 and IGC5323 showed higher leavening ability than Saccharomyces, specially after exposure to hyperosmotic stress of bread dough containing 20% sucrose and 2% salt added. In addition, Torulaspora and especially T. delbrueckii IGC5321 exhibited no loss of CO2 production capacity during freeze-thaw stress. Overall, these results appeared to indicate that Torulaspora cells are more tolerant than Saccharomyces to osmotic stress of bread dough. This trait correlated with a low invertase activity, a slow rate of trehalose mobilisation and the ability to respond rapidly to osmotic stress. Growth behaviour on high osmotic synthetic media was also examined. Cells of the IGC5321 strain showed intrinsic osmotolerance and ion toxicity resistance. However, T. delbrueckii IGC5323 exhibited a clear phenotype of osmosensitivity. Hence, this characteristic may not be essential or the only determinant for leavening ability in salted high sugar dough. PMID- 14533717 TI - Exopolysaccharide production by filamentous fungi: the example of Botryosphaeria rhodina. AB - One-hundred and five fungal strains, belonging to 46 different species, were screened for exopolysaccharide production. Phytopathogenicity and, in particular, inability to produce conidia, were physiological characteristics positively associated and correlated with the fungal ability to produce polysaccharides. Among the 29 positive strains, Botryosphaeria rhodina DABAC-P82 was the most interesting reaching, when grown on optimal nitrogen source and concentration (NaNO3 and 2.0 g l(-1), respectively) and culture medium pH (3.7), 17.7 g l(-1) of exopolysaccharide production after only 24 h of fermentation; yield and productivity were 0.69 g g(-1) and 0.73 g l(-1)h(-1), respectively. The purified polysaccharide was characterised as a homopolysaccharide of glucose with a molecular weight of 4.875 x 10(6) Da. Studies of structural analysis indicated the presence of beta-1,3 and beta-1,6 linkages; the EPS structure was very similar to that of scleroglucan. PMID- 14533718 TI - Six new methanol assimilating yeast species from wood material. AB - Ten yeast strains representing six hitherto unknown methanol utilizing yeast species were isolated from tree exudate, bark and rotten wood samples. Following the sequencing of the D1/D2 region of their large (26S) subunit rDNA, the four ascosporogenous species were assigned to the genus Pichia, and the two anascosporogenous ones to the genus Candida. Although genetically clearly separated, three of the four new Pichia species are phenotypically very similar to P. pini, and they can be differentiated only by minor physiological and morphological characteristics. The description is given for the six new species (C. suzukii, C. hungarica, P. trehaloabstinens, P. pilisensis, P. dorogensis and P. zsoltii). PMID- 14533719 TI - Competing interests: the importance of transparency. PMID- 14533720 TI - Is parental sense of coherence associated with child health? AB - Antonowsky's concept of sense of coherence (SOC) has, during recent years, gained increased attention as a salutogenic model on the relationship between health and disease. However, only sparse information connecting child chronic health conditions to parental SOC is yet available. This article presents results from a cross-sectional study of about 10,000 children aged 2-17 years in the five Nordic countries in 1996. Factors associated with parental SOC were analysed, with focus on child chronic health conditions. Sense of coherence was measured according to a short and condensed three-item instrument based on Antonovsky's original 29 item instrument. Overall, about 23% of parents in Nordic countries had a poor sense of coherence, the lowest proportion found among Icelandic parents. Compared to the higher social classes, poor SOC was more common in the lower social classes. The association of child chronic health conditions with parental poor SOC was found to be disability specific. Parents of children with diabetes, epilepsy or psychiatric/nervous problems had approximately 2-5 higher odds of having poor SOC compared to parents of children without a specific diagnosis. The overall effect of having a child with chronic health conditions was, however, low, lower than the effect of the parents' own health complaints. PMID- 14533721 TI - Visual disturbances in a population-based survey of 6962 subjects: the German National Health Examination Survey 1998. AB - BACKGROUND: Available prevalence estimates of visual disturbances (excluding blindness) in Germany are based on data from highly selective populations. This report describes the prevalence of visual disturbances and potential determinants based on the German National Health Examination Survey from 1998. METHODS: A population-based cross-sectional study of the non-institutionalized population in unified Germany. People aged 18-79 years were eligible and were contacted by a multi-mode approach. The response proportion was 61%, resulting in a sample of 7124 subjects who participated in the study. Visual disturbances were assessed by a self-administered questionnaire. RESULTS: Prevalance rates are higher among women in unified as well as in East- and West Germany. The higher overall prevalence rates among women is mostly driven by higher prevalance rates at ages 18-49 years, especially for shortsightedness. The prevalence rates are higher in West Germany than East Germany. Visual disturbances are more prevalent among the middle and upper social class than the lower social class. Above the age group 40 44, the prevalence of visual disturbances considerably increases, to approximately 100% in the age group 55 years or older for both sexes and in both parts of Germany. CONCLUSIONS: Nearly 100% of German adults aged 50-79 years have some degree of visual disturbance that requires refractive correction. Prevalence rates of visual disturbances are higher among people from West Germany, people of higher social status and among women. Uncorrected visual disturbances are most prevalent in the age group 18-34 years and more often among males and subjects of lower social status. PMID- 14533723 TI - Psychological distress one year after childbirth: a cross-cultural comparison between France, Italy and Quebec. AB - BACKGROUND: The aim of this study was to compare rates of psychological distress, one year after childbirth, between women living in France, Italy and Quebec (Canada). METHODS: Analysis was performed for a sample of 1663 mothers of one year-old children. Psychological distress rates, measured by the General Health Questionnaire and a measure of psychotropic drug use, were compared in bivariate and multivariate analysis. RESULTS: According to the GHQ scores, the psychological distress rate was significantly higher in Quebec (16% scoring over 5) than in France (11%) and Italy (9%). Differences between countries were most marked for women with a low level of education, who were more distressed in Quebec than elsewhere. However, psychotropic drug use was significantly more common in France than in Italy and Quebec. When the two indicators of psychological distress were combined (high GHQ score or use of psychotropic drugs), France and Quebec presented similar distress rates, both significantly different from Italy. CONCLUSION: Differences in the expression of distress may partly account for the observed differences in distress rate between countries. However, the social structure of each country probably plays a role in the development of psychological distress, possibly leading to differences in distress rate between countries. PMID- 14533722 TI - Psychosocial factors, lifestyle, and fetal growth: the added value of both pre- and post-natal assessments. AB - BACKGROUND: Psychosocial resources as well as lifestyle habits during pregnancy have been shown to effect the risk of having a small-for-gestational-age (SGA) child. Most previous studies are based on a single assessment of these exposures, which does not take into account the possibility of different effects during early and late stages of pregnancy. METHODS: The impact of psychosocial and lifestyle factors on the risk of giving birth to an SGA child (as measured by ultrasound) was examined among 747 nulliparous Swedish women who completed both a prenatal baseline, and a post-partum assessment. RESULTS: Those registering low social participation on both assessments showed increased risk of giving birth to an SGA infant (OR = 2.44 and 95% CI: 1.06-5.66), while at one assessment (OR = 1.70 and 95% CI: 0.74-3.91). Maternal smoking confirmed by both or one assessments yielded an OR = 2.72 and 95% CI: 1.37-5.39 and OR = 1.60 and 95% CI: 0.58-4.46, respectively. During early pregnancy, poor instrumental support, maternal smoking, or passive smoking yielded increased risks of SGA, adjusted for confounding (OR = 2.39 and 95% CI: 1.11-5.17; OR = 2.38 and 95% CI: 1.27-4.49; OR = 2.92 and 95% CI: 1.17-7.32, respectively). In late pregnancy, only maternal smoking yielded a significant association (OR = 2.34 and 95% CI: 1.24-4.41). CONCLUSION: Scheduling repeated assessments of psychosocial resources and lifestyle factors during pregnancy yielded additional information. The findings suggest that there can be differential effects of such exposures depending on gestational stage. This information is of importance when designing appropriate intervention strategies for maternal health services as well as for public health relevant policy formulation (e.g. regarding exposure to environmental tobacco during pregnancy). PMID- 14533724 TI - High coronary heart disease rates among Dutch women of the baby boom, born 1945 1959: age-cohort analysis and projection. AB - BACKGROUND: After a steep decline in older generations, coronary heart disease mortality is stagnating in female cohorts born after the Second World War. We analysed past trends and predicted future health care needs for coronary heart disease in the Dutch population. METHODS: A loglinear age-cohort model relates numbers of deaths and hospital admissions for coronary heart disease to sex, age, birth cohort and population size, and projects age-cohort changes over the future population. Population size, population forecasts and coronary heart disease mortality (period 1970-1999) are from vital statistics. Numbers of hospitalised acute coronary events are from the nationwide hospital register (period 1980 1999). RESULTS: Among men, the rate ratios of deaths and hospital admissions were, respectively, 0.21 (death) and 0.78 (survivors at discharge) in the cohorts born in the period 1948-1962 compared to the period 1918-1922. Among women, the same rate ratios were 0.41 and 1.89. The projection model predicts 22% less deaths from coronary heart disease and 22% more survivors of an infarction in 2015, among men. Among women, there will be 5% less deaths and 70% more survivors of an infarction, most of these being middle age members of the baby boom cohorts. CONCLUSIONS: Stagnating all-cause mortality is correlated with an upward trend in coronary heart disease risk in the female baby boomers. Heart health care needs among middle-aged women will increase sharply. These changes are correlated to high lung cancer mortality and high smoking rates in these cohorts. PMID- 14533725 TI - Increasing mortality among adults in Scotland 1981 to 1999. AB - BACKGROUND: The aim of the study was to describe the change in overall and cause specific mortality in Scotland between the early 1980s and late 1990s, with particular reference to the mortality experience of young adults. METHOD: The study was based on death records for Scottish residents. Changes in age and cause specific death rates between 1981-83, 1989-91 and 1997-99 were compared. RESULTS: Between 1981-83 and 1989-91 death rates in Scotland began to rise among young men aged 20-24 while for those aged over 25 rates declined. The greatest fall in rates was experienced at ages 40 to 59. When death rates during 1997-99 were compared to rates in 1989-91 this pattern had changed. During the 1990s death rates among 20 to 34-year-olds increased, with a slight rise at ages 35-44. At older ages overall mortality continued to decline but the greatest fall was at ages 60 and over. Trends among women shared similarities with men. For both men and women falls in mortality from heart disease, stroke, and cancers were being differentially offset by increases in other causes of death across all age groups. The causes of death that contributed to the increased death rate among young adults include to various degrees, suicides, drug deaths, alcohol and violence. CONCLUSION: In Scotland changes in mortality result from a complex combination of different trends in mortality from various causes of death. The rate of decline in mortality among men aged 59 and below is slowing down, and death rates among young men aged 15-44 are increasing. If these trends continue there is a suggestion that future death rates may begin to rise at older ages. PMID- 14533726 TI - Health promotion activities in annual reports of local governments: 'Health for All' targets as a tool for content analysis. AB - BACKGROUND: This article presents an instrument to study the annual reporting of health promotion activities in local governments within the three intervention municipalities of the Stockholm Diabetes Prevention Program (SDPP). The content of health promotion activities are described and the strengths, weaknesses and relevance of the method to health promotion discussed. METHOD: A content analysis of local governmental reports from 1995-2000 in three Swedish municipalities. A matrix with WHO's 38 'Health for All' (HFA) targets from 1991 was used when coding the local health promotion activities. RESULTS: There are many public health initiatives within the local governmental structure even if they are not always addressed as health promotion. The main focuses in the local governmental reports were environmental issues, unemployment, social care and welfare. CONCLUSIONS: Local governmental reports were found to be a useful source of information that could provide knowledge about the priorities and organizational capacities for health promotion within local authorities. Additionally the HFA targets were an effective tool to identify and categorize systematically local health promotion activities in the annual reports of local governments. Identifying local health promotion initiatives by local authorities may ease the development of a health perspective and joint actions within the existing political and administrative structure. This paper provides a complementary method of attaining and structuring information about the local community for developments in health promotion. PMID- 14533728 TI - European regulatory policies on medicines and public health needs. AB - The establishment of the EMEA has been a revolutionary step in the European pharmaceutical system. The 15 Member States of the European Union now share a common system for the evaluation of new medicinal products entering the European market. The decisions taken apply to the whole EU, with important implications for both industry and patients who may benefit from new therapies. The main immediate consequences of this system are: i) the time and effort saved by Member States in the evaluation of new drug applications; ii) more consistent and quicker availability of medicines in EU countries; iii) the establishment of a homogeneous regulatory policy throughout the EU. Public health has been presented as the fundamental concern of the EMEA, the mission statement of which is 'to promote the protection of human health ... and of consumers of medicinal products'. However, we note that there are some inconsistencies with this objective and the current system, such as those regarding drug trial requirements and the institutional location and financing of the EMEA. In this paper, some aspects of the new system are reviewed and consideration given as to how they relate to public health needs. Proposals are made for debate alternatives and improvements to the present system that would better respond to patients' health needs. PMID- 14533727 TI - Building integrated health systems in central and eastern Europe: an analysis of WHO and World Bank views and their relevance to health systems in transition. AB - BACKGROUND: Two questions are addressed. i) What are the views on health and health systems as expressed in the World Development Report 2000/2001 of the WB and the World Health Report 2000 and Health 21 of the World Health Organization, and how compatible are those views? ii) To what extent will compliance of CEEC and NIS with the WHO and WB recommendations result in health systems that produce maximum health for all by adequately addressing the needs of their populations? METHOD: The reports prepared by the World Bank and the World Health Organization were assessed against the theoretical framework of a needs-based public health approach. RESULTS: It is observed that the WHO and WB approaches are currently converging, although there remain differences in their respective focuses. The main merit of the WHO approach is its focus on performance and the systems approach towards health (care). The merit of the WB view is the integrated approach to health, education and poverty. It is argued that CEEC and NIS need to anticipate an ageing population and growing numbers of chronically ill. This calls for integrated health care systems and more integrated funding and payment systems. CONCLUSION: The recommendations provided in the WHR and the WDR with regard to integrated care and integrated financing remain rather abstract. Advisors of CEEC and NIS on health care reform and Western assistance projects should focus more on future needs, in order to avoid building health systems that consistently lag behind the needs of their populations. PMID- 14533729 TI - Legal access to needles and syringes/needle exchange programmes versus HIV counselling and testing to prevent transmission of HIV among intravenous drug users: a comparative study of Denmark, Norway and Sweden. AB - BACKGROUND: Countries have adopted different strategies to prevent the transmission of HIV among intravenous drug users. Legal access to needles and syringes/needle exchange programmes as part of such a strategy has been heavily debated. HIV counselling and testing has also been part of prevention strategies. The objective of this study was to discuss the effectiveness of legal access to needles and syringes/ needle exchange programmes versus HIV counselling and testing among intravenous drug users (IDUs) as part of HIV prevention strategies. METHODS: Differences in HIV prevention strategies in Denmark, Norway and Sweden among IDUs are described. Outcome variables of effectiveness were HIV incidence rates over time. These were estimated by back calculation methods from 1980 through 1996, using data from the national HIV and AIDS registers. RESULTS: A comparison of HIV prevention strategies in Denmark, Norway and Sweden suggests that a high level of HIV counselling and testing might be more effective than legal access to needles and syringes/needle exchange programmes. Sweden and Norway, with higher levels of HIV counselling and testing, have had significantly lower incidence rates of HIV among IDUs than Denmark where there was legal access to needles and syringes and a lower level of HIV counselling and testing. In Sweden there was no legal access to drug injection equipment. CONCLUSION: Promotion and accessibility of HIV counselling and testing among intravenous drug users should be considered in countries where such a strategy is not adopted or has low priority. PMID- 14533730 TI - Deprivation and AIDS in a southern European city: different patterns across transmission group. AB - OBJECTIVE: To analyse deprivation and AIDS among three AIDS transmission groups (men who have sex with men--MSM, heterosexuals, and intravenous drug users--IDUs) in Barcelona, Spain, during the period 1990-95. METHODS: This is an ecological study, the unit of analysis being the neighbourhoods. Included were AIDS cases residents in Barcelona. The association among AIDS rate and deprivation was studied using Spearman correlation coefficients and Poisson regression. RESULTS: For MSM, inner city neighbourhood residence meant a greater risk of AIDS; but lower educational level was inversely related with AIDS rates. For heterosexuals, variables related with AIDS rates were younger age, inner city areas and social unrest for women, and extreme poverty for men. Among UDIs variables related with AIDS were unemployment and social unrest for both sexes. CONCLUSION: The association between AIDS rates and deprivation differs across transmission groups in a southern European city. PMID- 14533731 TI - Development and validation of a short food list to assess the intake of total fat, saturated, mono-unsaturated, polyunsaturated fatty acids and cholesterol. AB - BACKGROUND: The aim of this study was the development and validation of a short list of food items to assess the intake of total fat, saturated, mono unsaturated, polyunsaturated fatty acids, and cholesterol. The short list should be able to correctly classify persons according to their intake. METHODS: A short list of 20 food items was selected out of 1009 seven-day food records by means of the variance-based method Max_r. This list was validated using data from a further 479 persons who completed seven-day food records (validation sample 1, VS1) as well as a food frequency questionnaire (validation sample 2, VS2). The intake of total fat, different fatty acids, and cholesterol from the complete VS1 (VS1(complete)) and from the complete VS2 (VS2(complete)), respectively, was computed. Further, the intake in VS1 (VS1(short)) as well as in VS2 (VS2(short)) using only the 20 food items on the short list were calculated. RESULTS: Pearson correlation coefficients between the intake calculated from the items on the short list and the nutrient intake calculated from the full instrument in VS1 and VS2, respectively, were r = 0.81-0.91. In a quartile's cross-classification 53.4 64.1% of the participants were assigned to the same quartile. When comparing VS1(complete) with VS2(short), neither correlation coefficients nor the cross classification differ much from the comparison of VS1(complete) with VS2(complete). CONCLUSIONS: The short list shows good results in both validation samples. Thus, the short list can assess the variability of fat intake and classify persons according to their intake. PMID- 14533732 TI - Parental education on passive smoking in infancy does work. AB - BACKGROUND: Passive smoking is harmful to young children. A protocol has been developed to allow health care workers to communicate with parents about preventing passive smoking. The main message was to refrain from smoking in the presence of the child. The aim of the study was to assess the effectiveness of this education programme. METHOD: The prevalence of smoking in the presence of infants aged 0-10 months was compared before and after the implementation of the education programme. National samples of mothers completed questionnaires in 1996 (n = 1,129) and in 1999 (n = 2,534). Questions were asked about smoking in the living room in the presence of infants, and about parental smoking, and background characteristics. RESULTS: The prevalence of passive infant smoking decreased from 41% to 18%. The adjusted odds ratio for passive infant smoking in 1999 compared to 1996 was 0.34 (0.26-0.44) when none of the parents smoked, 0.19 (0.14-0.27) when one of the parents smoked, and 0.30 (0.20-0.44) when both parents smoked. CONCLUSION: The implementation of this health education programme seems to have been very successful in reducing passive smoking in children. Implementation of similar health education programmes in other countries is recommended. PMID- 14533733 TI - Tobacco- and alcohol-attributable mortality and years of potential life lost in Germany. AB - AIM: To compare the ages of death caused by tobacco smoking and alcohol risk drinking. METHODS: Smoking rates from the largest population survey, alcohol drinking data from the National Health Survey and data from the vital statistics from Germany are used and attributable fractions computed. RESULTS: Alcohol attributable deaths occurred at the youngest age, followed by tobacco- plus alcohol-attributable cases, whereas death cases attributable to tobacco smoking only occur latest. CONCLUSION: The overlap in the two substance-use behaviours has to be taken into account when considering attributable mortality data. PMID- 14533735 TI - 'Salicylic acid deficiency' has important public health implications. PMID- 14533734 TI - Cigarette smoking, alcohol consumption, and serum lipid profile among medical students in Greece. AB - BACKGROUND: The lack of data regarding health habits of medical students in Greece prompted a cross-sectional study to assess tobacco use, alcohol consumption and serum lipoprotein levels among students in the University of Crete School of Medicine. METHODS: A self-administered questionnaire on smoking and alcohol consumption was distributed to third-year medical students for twelve consecutive years (1989-2000). A total of 849 students (462 males, 387 females) participated in the survey. Biochemical measurements were taken and multi-variant analysis of the data was performed. RESULTS: The prevalence of smoking among males and females was 33.2% (N = 150) and 28.4% (N = 108), respectively (mean cigarette consumption 13/day). As many as 349 males (77.2%) and 220 females (58.0%) reported consuming alcohol on a regular basis. The prevalence of low HDL cholesterol (< 0.9 mmol/l) was 14.5% in males and 5.1% in females, and of high LDL-cholesterol levels (> 4.1 mmol/l) in 11.1% of male and 5.5% of female participants. Smoking was related to higher triglyceride (p = 0.032), and lower HDL-cholesterol (p = 0.037) serum levels. Total cholesterol, LDL-cholesterol, and the TC/HDL-cholesterol ratio were strongly related with the level of smoking (p = 0.006, p = 0.008, and p = 0.006 respectively). CONCLUSIONS: The results document a high prevalence of smoking among physicians-to-be in Greece. Tobacco use was strongly associated with a lipid profile predisposing to increased risk for cardiovascular disease. Health promotion programmes should therefore be instituted not only during the first years of medical studies, but rather at a much earlier stage in life. PMID- 14533736 TI - Postscript, commentary ministerial challenges: post-conflict, post-election issues in Kosovo's health sector. PMID- 14533737 TI - Tight junction-related structures in the absence of a lumen: occludin, claudins and tight junction plaque proteins in densely packed cell formations of stratified epithelia and squamous cell carcinomas. AB - Tight junctions (TJs), hallmark structures of one-layered epithelia and of endothelia, are of central biological importance as intramembranous "fences" and as hydrophobic "barriers" between lumina represented by liquid- or gas-filled spaces on the one hand and the mesenchymal space on the other. They have long been thought to be absent from stratified epithelia. Recently, however, constitutive TJ proteins and TJ-related structures have also been identified in squamous stratified epithelia, including the epidermis, where they occur in special positions, most prominently in the uppermost living epidermal cell layer, the stratum granulosum. Much to our surprise, however, we have now also discovered several major TJ proteins (claudins 1 and 4, occludin, cingulin, symplekin, protein ZO-1) and TJ-related structures in specific positions of formations of epithelium-derived tissues that lack any lumen and do not border on luminal or body surfaces. Using immunohistochemistry and electron microscopy we have localized TJ proteins and structures in peripheral cells of the Hassall's corpuscles of human and bovine thymi as well as in specific central formations of tumor nests in squamous cell carcinomas, including the so-called "horn pearls". Such structures have even been found in carcinoma metastases. In carcinomas, they often seem to separate certain tumor regions from others or from stroma. The structural significance and the possible functional relevance of the locally restricted synthesis of TJ proteins and of the formations of TJ-related structures are discussed. It is proposed to include the determination of the presence or absence of such proteins and structures in the diagnostic program of tumor pathology. PMID- 14533738 TI - Targeting of the human adrenoleukodystrophy protein to the peroxisomal membrane by an internal region containing a highly conserved motif. AB - In this study we addressed the targeting requirements of peroxisomal ABC transporters, in particular the human adrenoleukodystrophy protein. This membrane protein is defective or missing in X-linked adrenoleukodystrophy, a neurodegenerative disorder predominantly presenting in childhood. Using adrenoleukodystrophy protein deletion constructs and green fluorescent protein fusion constructs we identified the amino acid regions 1-110 and 67-164 to be sufficient for peroxisomal targeting. However, the minimal region shared by these constructs (amino acids 67-110) is not sufficient for peroxisomal targeting by itself. Additionally, the NH2-terminal 66 amino acids enhance targeting efficiency. Green fluorescent protein-labeled fragments of human peroxisomal membrane protein 69 and Saccharomyces cerevisiae Pxa1 corresponding to the amino acid 67-164 adrenoleukodystrophy protein region were also directed to the mammalian peroxisome. The required region contains a 14-amino-acid motif (71-84) conserved between the adrenoleukodystrophy protein and human peroxisomal membrane protein 69 and yeast Pxa1. Omission or truncation of this motif in the adrenoleukodystrophy protein abolished peroxisomal targeting. The single amino acid substitution L78F resulted in a significant reduction of targeting efficiency. The in-frame deletion of three amino acids (del78-80LLR) within the proposed targeting motif in two patients suffering from X-linked adrenoleukodystrophy resulted in the mislocalization of a green fluorescent protein fusion protein to nucleus, cytosol and mitochondria. Our data define the targeting region of human adrenoleukodystrophy protein containing a highly conserved 14-amino-acid motif. PMID- 14533739 TI - Membrane targeting of p115 phosphorylation mutants and their effects on Golgi integrity and secretory traffic. AB - The cytosolic phosphoprotein p115 is required for ER to Golgi traffic and for Golgi reassembly after mitosis. In cells, p115 is localized to ER exit sites, ER Golgi Intermediate Compartment (ERGIC) and the Golgi, and cycles between these compartments. P115 is phosphorylated on serine 942, and this modification appears to control p115 association with membranes. P115 is likely to function by reversibly interacting with effector proteins, and in the Golgi, two proteins, GM130 and giantin, have been shown to bind p115. The GM130-p115 and the giantin p115 interactions are enhanced by p115 phosphorylation. Phosphorylation appears to be essential for p115 function, since substitutions of serine 942 abolish p115 ability to sustain cisternal reformation in an in vitro assay reconstituting Golgi reassembly after mitosis. Here, we explored how phosphorylation of p115 affects its intracellular targeting to distinct cellular compartments, and its function in secretory traffic. We generated phosphorylation mutants of p115 and tested their ability to target to ER exit sites, ERGIC and the Golgi. In addition, we explored whether expression of the mutants causes disruption of Golgi structure and perturbs ER-Golgi traffic of a VSV-G cargo protein. PMID- 14533740 TI - Role of apoptosis and mitosis during human eye development. AB - The spatial and temporal distribution as well as ultrastructural and biochemical characteristics of apoptotic and mitotic cells during human eye development were investigated in 14 human conceptuses of 4-9 postovulatory weeks, using electron and light microscopy. In the 5th developmental week, apoptotic and mitotic cells were found in the neuroepithelium of the optic cup and stalk, being the most numerous at the borderline between the two layers of the optic cup, and at the place of transition of the optic cup into stalk. They were also found at the region of detachment of the lens pit from the surface ectoderm. In the later developmental stages (the 6th-the 9th week), apoptotic and mitotic cells were observed in the neural retina and the anterior lens epithelium. Throughout all stages examined, mitotic cells were found exclusively adjacent to the lumen either of the intraretinal space or the optic stalk ventricle, or were restricted to the superficial epithelial layer of the lens primordium. Unlike mitotic cells, apoptotic cells occurred throughout the whole width both of the neuroepithelium and the surface epithelium. Ultrastructurally, apoptotic cells were characterised by round- or crescent-shaped condensations of chromatin near the nuclear membrane, while in the more advanced stages of apoptosis by apoptotic bodies. The distribution of caspase-3-positive cells coincided with the location of apoptotic cells described by morphological techniques indicating that the caspase-3 dependent apoptotic pathway operates during the all stages of human eye development. The location of cells positive for anti-apoptotic bcl-2 protein was in accordance with the regions of eye with high mitotic activity, confirming the role of bcl-2 in protecting cells from apoptosis. In the earliest stage of eye development, apoptosis and mitosis might be associated with the sculpturing of the walls of optic cup and stalk, while high mitotic activity along the intraretinal space and optic stalk ventricle indicates its role in the gradual luminal closure. These processes also participate in the detachment of the lens pit epithelium from the surface ectoderm as well as in further closure of the lens vesicle. Later on, both processes seem to be involved in the neural retina differentiation, lens morphogenesis and secondary lens fibre differentiation. PMID- 14533741 TI - Posttranslational isomerization of a neuropeptide in crustacean neurosecretory cells studied by ultrastructural immunocytochemistry. AB - Isomerization of the third amino acid residue (a phenylalanine) of crustacean hyperglycemic hormone (CHH) has been previously reported to occur as a late step of hormone precursor maturation in a few neurosecretory cells in the X-organ sinus gland complex of the crayfish Orconectes limosus. In the present report, using conformation-specific antisera combined with immunogold labeling, we have studied, at the ultrastructural level, the distribution of L- and D-CHH immunoreactivity in CHH-secreting cells of the crayfish Astacus leptodactylus. Two CHH-secreting cell populations were observed, the first one (L-cells), the most numerous, exhibited only labeling for L-CHH. In the second one (D-cells), four secretory granule populations were distinguished according to their labeling: unlabeled, either L- or D- exclusively or both L- and D-granules. Labeling quantification by image analysis in D-cells showed a marked increase in D-labeling from the cell body to the axon terminal. However some L- and mixed granules remain in axon terminals. Our results demonstrate that Phe3 isomerization of CHH occurs within the secretory granules of specialized neurosecretory cells and progresses as the granules migrate along the axonal tract. The observation that not all the CHH synthesized is isomerized, and the great variability in the proportion of L- and D-immunoreactivity in granules in every cell region may suggest an heterogeneous distribution of the putative enzyme involved in Phe3 isomerization, a peptide isomerase, within the secretory pathway. PMID- 14533742 TI - Creating a coherent set of indicators to monitor health across Europe: the Euro REVES 2 project. AB - The Euro-REVES 2 project, 'Setting up of a coherent set of health expectancies for the European Union', was begun in 1998 under the European Health Monitoring Programme with the aim of selecting a concise set of instruments to simultaneously monitor mortality and the different facets of health. An in-depth analysis of the current health survey instruments in Europe together with a review of past research, found that, although harmonization in instruments appeared to exist superficially, major differences existed. Four instruments have been recommended (where necessary using existing instruments with modifications suggested by the research literature) covering physical and sensory functional limitations, activity restriction, self-perceived health and mental health. Additionally a new global activity limitation indicator (GALI) has been developed. These instruments are firmly anchored to past research and the health concepts behind the indicators and their relevance to policy and guidelines for implementation are explicitly made. The second phase of the project will recommend further instruments, leading to health expectancies that cover all the conceptual framework of population health measurement. This will allow assessment of health inequalities between the European Union countries, an appreciation of the causes and the production of profiles for each country in terms of the various facets of health. PMID- 14533743 TI - Assessing comparability of dressing disability in different countries by response conversion. AB - BACKGROUND: Comparability of health data is a major challenge within the context of the Health Monitoring Programme of the European Commission. A common problem in surveys is that many variations of essentially the same question exist. METHODS: Response conversion is a new method for improving comparability by scaling the data onto a common scale. Comparisons between member states can then be made in terms of the common scale. A first step is the construction of a conversion key. This is a relatively complex activity, but needs to be done only once. The second step is the actual data transformation. This is simple, and can be repeatedly done on a routine basis as new information arrives. Construction of the key is only possible if enough overlapping information can be found. RESULTS: The method is illustrated for dressing disability from five European countries. Differences occur between countries, between sexes and between age groups. These were similar in magnitude. CONCLUSION: Response conversion is a new method for enhancing comparability among existing data. Conversion can only be done if a key is available. More work is needed to establish the technique. Future implications within the Health Monitoring Programme are discussed. PMID- 14533744 TI - International monitoring of musculoskeletal complaints: a need for consensus. AB - Musculoskeletal pain is the complaint most frequently reported in health interview surveys. There is a confusing magnitude of names given to musculoskeletal pain complaints reflecting diverging opinions and a lack of consensus of what it is all about. This article discusses how to monitor the occurrence of unspecified musculoskeletal complaints and possible sources of information are presented. The main focus of the article is on how to include musculoskeletal complaints in health interview surveys, and a standard question to be included in all general health questionnaires is recommended. PMID- 14533745 TI - European food availability databank based on household budget surveys: the Data Food Networking initiative. AB - BACKGROUND: There is a need in Europe for sources of dietary data that would provide a regular flow of comparable nutrition information. In this context, the Data Food Networking (DAFNE) project has been developing a cost-effective food databank that allows monitoring of food availability within and between countries. METHODS: The DAFNE project has developed a common classification system for the food and socio-demographic variables recorded in the national household budget surveys. Daily individual food availability was estimated for each participating country and basic descriptive statistics were used to depict the nutritional habits of the populations. RESULTS: The DAFNE databank is freely accessible through DafneSoft (www.nut.uoa.gr). Three figures, based on Greek, Norwegian and UK data respectively, were selected to indicatively present the potential of the DAFNE data in order to monitor changes in the availability of 12 food groups over time; to study the effect of education in the total added lipid availability; and to follow time changes in fresh vegetable availability between manual and non-manual households. CONCLUSION: The HBS data allows the highlighting of issues related to nutrition surveillance. The application of the DAFNE methodology in developing a databank of standardized dietary data offers a realistic option for monitoring dietary habits in Europe. Benefiting from this potential, the European Union may additionally consider the undertaking of a pan European individual dietary survey, in a sub-sample of the household budget population. PMID- 14533747 TI - Child health indicators for Europe: a priority for a caring society. AB - BACKGROUND: Measurement of children's health is important for two reasons: first, because young people are citizens in their own right, yet largely unable to act as self-advocates, particularly at the population level; and second, because their health determines the health of the future population. Indicators based on measurements of child health are important for identifying progress, problems and priorities, changes over time, and newly emergent issues. The European Community Health Monitoring Programme (HMP) is a comprehensive programme to develop and implement a set of national-level indicators. The Child Health Indicators of Life and Development (CHILD) project is the only population group-specific project, seeking to determine a holistic set of measures. METHODS: The project endeavoured to address all aspects of child health and its determinants, balancing positive and negative aspects. It undertook a structured search of published evidence to seek to identify, and validate, indicators of health and illness, health determinants and challenges to health, quality of healthcare support and health promoting national policies. A systematic approach was used in identifying valid indicators, and in assembling a balanced composite list. All ages from infancy to adolescence were covered. RESULTS: The project's final report identifies 38 core desirable national indicators, citing purpose and evidence for each. Of equal importance, it also identifies 17 key child health topics on which further research work is needed in order to identify and validate indicators appropriate across different national settings. PMID- 14533746 TI - PERISTAT: indicators for monitoring and evaluating perinatal health in Europe. AB - BACKGROUND: The PERISTAT project aimed to develop an indicator set for monitoring and describing perinatal health in Europe. The challenge was to define indicators that cover common concerns and have the same meaning within the different European health care systems. METHODS: PERISTAT included i) a review of existing recommendations on perinatal health indicators, ii) a DELPHI consensus process with a scientific advisory committee composed of a clinician and an epidemiologist or statistician from each European member state as well as with a panel of midwives, and iii) a study of the availability of national statistics to construct recommended indicators. This article describes the first two components. RESULTS: The review identified 10 international and 13 national recommended indicator sets. It also included indicators routinely compiled by WHO, EUROSTAT and OECD. Because of the methodological limits to using existing indicators for European comparisons, a high priority was placed on improving indicators already collected. Using the DELPHI method based on the results of the review, the scientific committee achieved a consensus on ten core and 23 recommended indicators, including 12 requiring further development. CONCLUSIONS: The PERISTAT project was successful in identifying a set of indicators, which drew on and consolidated previous work. Consensus was not achieved on precise indicators in areas where uncertainty about appropriate indicators was high, although areas were targeted for future development. Finally, the feasibility study, which is in progress, is an essential part of the project, since it will enable member states to evaluate their capacity to produce these indicators. PMID- 14533748 TI - Health indicators for people with intellectual disabilities: a European perspective. AB - People with intellectual disabilities make up about 1% of the population of Europe. As trends toward community life advance, they have become more visible and more likely to access generic health systems. Yet evidence suggests that there are striking disparities between the health of this group and that of the general population. Increased longevity means that adults in this group expect to live longer lives and thus to encounter age-related risks for various health conditions. The 'Pomona' project, funded by the EU Health Monitoring Unit, aims to develop a set of health indicators for people with intellectual disabilities. It will build on the work accomplished by the team developing ECHI--European Community Health Indicators. This article outlines the rationale for the project, key elements in its implementation and expected outcomes. PMID- 14533749 TI - European Union diabetes indicators: fact or fiction? AB - Diabetes mellitus is one of the major causes of morbidity and mortality in EU/EFTA countries. Monitoring risk factors for diabetes and its complications will offer the possibility to evaluate the development in time as well as the influence of possible interventions. In this investigation a list with core and secondary indicators is proposed. Availability of these indicators and their data sources is discussed. An important variability of data sources is used in EU/EFTA countries, interfering with the comparability of the outcome. Further harmonisation as well as continuous evaluation of data sources will be necessary to provide reliable tools to monitor diabetes mellitus and its outcome on a routine basis. PMID- 14533750 TI - Coronary and cerebrovascular population-based registers in Europe: are morbidity indicators comparable? Results from the EUROCISS Project. AB - BACKGROUND: The EUROCISS Project (European Cardiovascular Indicators Surveillance Set), as part of the Health Monitoring Programme financed by the European Commission, has been implemented to develop health indicators and recommendations for the monitoring of cardiovascular diseases (CVDs). Morbidity data are rarely available in the different countries and when available, they are very rarely comparable. The aims of this paper are to list the existing population-based registers of acute myocardial infarction (AMI) and stroke in Europe, describe their methodology, and discuss their comparability. METHODS: using a questionnaire a comprehensive and updated picture on available sources of information, data, indicators, and methods were collected for population-based registers. The information requested generally included: the studied disease; the scope of the study (geographical area, temporal duration, age range, population); adopted methodologies (case definition, ICD coding for mortality and hospital discharge records, linkage and validation methods); morbidity indicators (attack rate, incidence, prevalence, case fatality rate). RESULTS: Belgium, Denmark, Finland, France, Germany, Italy, Norway, Spain, and Sweden have ongoing population-based registers for AMI. Denmark, Finland, France, Germany, Italy, Norway, and Sweden have ongoing population-based registers for stroke. Selection procedures of events, differences in age range, different validation procedures and methods make the results from these registers difficult to compare. CONCLUSIONS: Population-based registers provide the best indicators for AMI and stroke, such as attack rate and case fatality. Registers cover large samples of the population, usually regions or large municipalities. The comparability of data across countries depends on standardization, case definition, completeness, proper linkage, common diagnostic criteria and validation procedures. Given the high burden of AMI and stroke, efforts are needed in implementing registers in all European countries. PMID- 14533751 TI - The comprehensive cancer monitoring programme in Europe. AB - BACKGROUND: There continue to be major public health challenges arising from the increasing cancer burden in Europe. Drawing upon expertise from other European centres and networks, the Comprehensive Cancer Monitoring Programme in Europe project (CaMon) provides a central information resource of the cancer profile in European populations. METHODS: The cancer indicators fundamental to disease monitoring in Europe are illustrated in terms of definitions and availability. Where necessary data are supplemented by estimates, in order to make available cancer data to individuals and institutions in all Member and Applicant countries of the European Union (EU). The relevant methodologies are discussed. Finally, a major ongoing project examining time trends of cancer incidence and mortality in 38 European countries is described. RESULTS: In the European Union, there were approximately 1.6 million new cases of cancer according to the latest year available, and approximately, one million cancer deaths. About 2.6 million new cases of cancer, and 1.6 million deaths were estimated in Europe. Lung cancer is the most common cancer in Europe and together with cancers of the colon and rectum and female breast represent approximately 40% of new cases in Europe. CONCLUSION: The statistics generated by the project on cancer incidence, mortality, survival and prevalence, together with time trends and projections will be regularly updated and made available to a European Commission, and to a Community-wide audience via the CaMon website and via other means of dissemination, such as peer-reviewed journals. PMID- 14533752 TI - Evaluation of Health Interview Surveys and Health Examination Surveys in the European Union. AB - BACKGROUND: The project 'Health surveys in the EU: HIS and HIS/HES evaluations and models' aims to assess the coverage of specific health and health-related areas in national and international surveys by reviewing and evaluating surveys, their methods and comparability, and by recommending appropriate survey designs and methods. METHODS: As basis for the evaluation, the project developed a health survey database. At present, Health Interview Surveys (HIS) and Health Examination Surveys (HES) from 18 Western European countries as well as from Canada, Australia and the USA are included. RESULTS: National HISs have been carried out regularly in almost all Western European countries. The HIS may consist of short health sections or modules within multi-purpose surveys or lengthy health interviews with several questionnaires. National HESs with a comprehensive focus have been conducted at regular or irregular intervals in five countries. The HES may comprise an interview and a few measurements or a comprehensive health examination. Sampling frames, fieldwork, quality control procedures and response rates vary greatly. Differences between measurement instruments used, in the wording of questions and in examination protocols reduce the comparability of many findings. CONCLUSION: The Internet based HIS/HES database allows for a quick reference and comparison of methods and instruments used in national health surveys. It illustrates the need for improving comparability. Collaboration and co-ordination is needed to promote comprehensive health monitoring supporting the development of national and European-level health policy. PMID- 14533753 TI - The Hospital Data Project: comparing hospital activity within Europe. AB - BACKGROUND: The ability to measure and compare hospital activity between EU member states is important for policy, planning, financing and assessment of population health. Earlier initiatives in this area have been largely directed at standardising high-level indicator definitions without proper account of differences in health systems and health information systems. The Hospital Data Project (HDP) develops a methodology for improved comparability of hospital inpatient and day case activity data across Europe and produces a pilot common data set. All EU members, Iceland and the World Health Organisation are participants. METHODS: The approach comprises a detailed inventory of patient level hospital data, identification of common areas, specification of data transformations and production of pilot data sets and metadata in a common format. An expert group developed a new diagnosis shortlist based on ICD-10. The project takes account of current work in the area of health care and morbidity indictors and applies the functional specification of health systems developed by the OECD. RESULTS: Seventeen countries have submitted data and metadata in the common format for a single year. Data on inpatients and day cases are classified by age, gender, diagnosis and type of admission. Numbers of hospital discharges, mean and median lengths of stay and population rates are reported. Test data on selected hospital procedures has also been collected. The full data set contains approximately 500,000 records, and software has been designed to facilitate validation and use. CONCLUSION: Results to date are promising. It is a first step in a complex area, and further work is required to extend and refine this approach. PMID- 14533754 TI - Health monitoring in sentinel practice networks: the contribution of primary care. AB - BACKGROUND: The health monitoring programme of the European Commission has proposed a set of health indicators whereby the health status of member states can be measured. As part of that programme we considered how primary care might contribute relevant data. METHODS: Using a questionnaire distributed to personal contacts and health authorities, we investigated the activities of sentinel practice networks and sought opinions on the place of primary care as a provider of information on health indicators. Studies on the prevalence of diabetes mellitus and on the incidence of chickenpox were undertaken within selected networks. RESULTS: 33 networks were found who provided relevant information on a timely and continuing basis. Contributions varied; some were limited to monitoring influenza but others recorded morbidity data from every consultation. Recording methods ranged from the paper based to fully automated systems in which all morbidity was coded electronically at data entry. The study of diabetes mellitus showed less variation between national networks than currently suggested on the WHO database. For chickenpox we estimated the incidence of cases not presenting to general practitioners ranged between 3 and 27%. CONCLUSIONS: Information on health indicators needs to come from the place where relevant care is delivered; for many conditions that is from primary care. It can be delivered from appropriately resourced practices where the population is defined, the practice populations are nationally representative and data collection is automated. PMID- 14533755 TI - European Emergency Data Project (EED Project): EMS data-based health surveillance system. AB - Emergency Medical Services (EMS) constitute a unique component of health care at the interface between primary and hospital care. EMS data within the pre-hospital setting represents an unparalleled source of epidemiological and health care information that have so far been neglected for public health monitoring. The European Emergency Data Project (EED Project) thus intends to identify common indicators for European EMS systems and to evaluate their suitability for integration into a comprehensive public health monitoring strategy. The article provides a brief overview on objectives and methodology in the form of a progress report. PMID- 14533756 TI - Work-related health monitoring in Europe from a public health perspective. AB - BACKGROUND: In contrast to the enormous importance of work to the life of humans and societies, the working environment has so far played only a minor role in health monitoring. However, increasingly it is realized that work has a strong public health impact and therefore is also a cost factor to modern societies. The aim of the project WORKHEALTH, which is currently being carried out under the EU health monitoring programme, is the establishment of indicators for work-related health monitoring in Europe from a public health perspective. WORKING PROCEDURE: The work will be carried out by means of three work packages: a synopsis of existing work-related indicator sets together with the identification of areas still to be developed, resulting specification of new indicators for work-related health monitoring, and the development of operational definitions for these indicators. The project includes strong links to the fields of occupational health and safety, public health, social insurance and labour inspectorates to include demands from, as well as to disseminate results to, these different institutions also involved in aspects of work-related health monitoring. EXPECTED RESULTS AND CONTRIBUTIONS: Expected results will be indicators for work-related health monitoring that have been developed jointly with the participation of all Member States and different stakeholders within this field, and which permit comparable and reliable data at a national as well as European level. PMID- 14533757 TI - EURO-MED-STAT: monitoring expenditure and utilization of medicinal products in the European Union countries: a public health approach. AB - BACKGROUND: There is uncertainty about the level of utilization and expenditure for medicines in the European Union (EU), making assessment of their impact on public health difficult. Our aim is to develop indicators to monitor price, expenditure and utilization of medicinal products in the EU, so as to facilitate comparisons. METHODS: There are four major tasks. Task 1: To catalogue data sources and available data in each EU Member State. Task 2: To assess the reliability and comparability of data among the EU Member States by ATC/DDD on country coverage, reimbursement, prescriptions, price category (e.g. wholesale, hospital, retail) and private versus public spending. Task 3: To develop Standard Operating Procedures for data management and to define clearly the proposed indicators in terms of objective, definition, description, rationale, and data collection. Task 4: To pool, compare and report the validated data according to the established indicators, using cardiovascular medicines as an example. RESULTS: Preliminary results from Tasks 1 and 2 are available and demonstrate the methodological difficulties in comparing data from different countries. Multiple data sources must be used. These cover different populations, and refer to different prices or costs. Nevertheless, useful data can be derived, illustrated by the example of lipid lowering medicines. The data shows that only five products are commonly available in all countries. Even when a medicine is available in all countries, there may be substantial differences in packages, which can hinder comparison. Data on utilization of statins shows high usage in Scandinavian countries and least in Italy. CONCLUSION: The preliminary results of EURO-MED-STAT show wide differences in availability, and use of medicines across Europe that may have substantial implications for public health. PMID- 14533758 TI - The ECHI project: health indicators for the European Community. AB - BACKGROUND: Within the EU Health Monitoring Programme (HMP), the ECHI project has proposed a comprehensive list of 'European Community Health Indicators'. METHODS: In the design of the indicator set, a set of explicit criteria was applied. These included: i) be comprehensive and coherent, i.e. cover all domains of the public health field; ii) take account of earlier work, especially that by WHO-Europe, OECD and Eurostat; and iii) cover the priority areas that Member States and Community health policies currently pursue. Flexibility is an important characteristic of the present proposal. In ECHI, this has been emphasized by the definition of 'user-windows'. These are subsets from the overall indicator list, each of which should reflect a specific user's requirement or interest. RESULTS: The proposed indicators are, in most cases, defined as generic indicators, i.e. their actual operational definitions have not yet been attempted. This work has been, and is being carried out to a large part by other projects financed under the HMP, which cover specific areas of public health or areas of data collection. Apart from indicators covered by regularly available data, indicators (or issues) have been proposed for which data are currently difficult to collect but which from a policy point of view would be needed. CONCLUSION: All this points to the fact that establishing an indicator list which is actually used by Member States is a continuously developing process. This process is now continued by the first strand of the new EU Public Health Action Programme. PMID- 14533759 TI - Advancing a theoretical model for public health and health promotion indicator development: proposal from the EUHPID consortium. AB - This paper discusses the work of the EUHPID Project to develop a European Health Promotion Monitoring System based on a common set of health promotion indicators. The Project has established three working groups to progress this task--health promotion policy and practice-driven, data-driven and theory-driven. The work of the latter group is reviewed in particular. EUHPID has taken a systems theory approach in order to develop a model as a common frame of reference and a rational basis for the selection, organization and interpretation of health promotion indicators. After reviewing the strengths and weaknesses of those health promotion models currently proposed for indicator development, the paper proposes a general systems model of health development, and specific analytical, socio-ecological models related to public health and health promotion. These are described and discussed in detail. Taking the Ottawa Charter as the preferred framework for health promotion, the socio-ecological model for health promotion adopts its five action areas to form five types of systems. The structure and processes for each of these five systems are proposed to form the basis of a classification system for health promotion indicators. The paper goes on to illustrate such a system with reference to indicators in the workplace setting. The EUHPID Consortium suggest that their socio-ecological model could become a common reference point for the public health field generally, and offer an invitation to interested readers to contribute to this development. PMID- 14533760 TI - The Euphin, the telematics support for public health in the EU. PMID- 14533761 TI - Cancer control in Europe: a proposed set of European cancer health indicators. PMID- 14533763 TI - New recommendations on anthelmintic use in sheep. PMID- 14533762 TI - Health indicators in the European Regions. PMID- 14533764 TI - Positives and negatives of pet ownership. PMID- 14533766 TI - Expression of cytokine mRNA in canine anal furunculosis lesions. AB - The pattern of expression of cytokine mRNA in the lesions of anal furunculosis was evaluated in tissue biopsies from 15 dogs, and compared with the pattern in control skin samples from 24 dogs, by reverse-transcriptase PCR using canine cytokine-specific primers and a semi-quantitative multiplex PCR assay. Interleukin-2 (IL-2) was detected in 11 of the 15 affected dogs but in only one of the controls, and interferon-gamma was detected in 14 of the affected dogs but none of the controls. In contrast, IL-4 was detected only in one of the affected dogs. Increased expression of mRNA for IL-1beta, IL-6, tumour necrosis factor alpha, IL-8, IL-10 and transforming growth factor beta1 was detected in the biopsies from the lesions of anal furunculosis relative to the control tissues (P < 0.05). PMID- 14533765 TI - Prevalence of faecal excretion of verocytotoxigenic Escherichia coli O157 in cattle in England and Wales. AB - During the decade to 1999, the incidence of human infections with the zoonotic pathogen verocytotoxin-producing Escherichia coli O157 (VTEC O157) increased in England and Wales. This paper describes the results of a survey of 75 farms to determine the prevalence of faecal excretion of VTEC O157 by cattle, its primary reservoir host, in England and Wales. Faecal samples were collected from 4663 cattle between June and December 1999. The prevalence of excretion by individual cattle was 4.2 per cent (95 per cent confidence interval [CI] 2.0 to 6.4) and 10.3 per cent (95 per cent CI 5.8 to 14.8) among animals in infected herds. The within-herd prevalence on positive farms ranged from 1.1 to 51.4 per cent. At least one positive animal was identified on 29 (38.7 per cent; 95 per cent CI 28.1 to 50.4) of the farms, including dairy, suckler and fattening herds. The prevalence of excretion was least in the calves under two months of age, peaked in the calves aged between two and six months and declined thereafter. The phage types identified most widely were 4, 34 and 2, which were each found on six of the 29 positive farms. PMID- 14533767 TI - Evaluation of biochemical methods for estimating the sodium intake of dairy cows. AB - Samples of blood and urine were taken from 334 dairy cows in 29 herds, and the concentrations of sodium and potassium were measured in the serum and urine. The herds were split into five groups according to the amount of supplementary salt they were given: three groups given 10 to 20, 30 to 50 or 70 to 100 g salt per day, and two groups fed an ad libitum supply, given either in bowls or in the form of salt blocks, which were replaced either regularly or irregularly. The groups which received 70 to 100 g of salt daily or were supplied regularly ad libitum had significantly higher urinary sodium concentrations than the other groups. The group receiving 10 to 20 g a day had a significantly higher ratio of potassium:sodium in their urine than all the other groups, in which the ratio decreased as the level of supplementary salt increased. There were wider differences between the groups in terms of the urinary potassium:sodium ratio than in terms of the urinary sodium concentration but less variability within each group. PMID- 14533768 TI - Intradural epithelial cyst in a cat. PMID- 14533769 TI - Collection of wild boar semen by electroejaculation. PMID- 14533770 TI - Isolation of Neospora caninum from a blind calf in Parana, southern Brazil. PMID- 14533771 TI - Spontaneous cholelithiosis in a hogget. PMID- 14533772 TI - Artificial insemination with frozen-thawed spermatozoa in dogs. PMID- 14533773 TI - Veterinary surveillance and client consent. PMID- 14533774 TI - Lay TB testing. PMID- 14533775 TI - Lack of PCV-2 infection in non-porcine species in Spain. PMID- 14533776 TI - Potential treatment for canine spinal cord injury. PMID- 14533777 TI - Piroplasmosis in a badger from Turkey. PMID- 14533778 TI - Cerebral amyloid angiopathies: a pathologic, biochemical, and genetic view. AB - Amyloid deposition can take place in the walls of arteries, arterioles, and, less often, capillaries and veins of the central nervous system, a phenomenon known as cerebral amyloid angiopathy (CAA). The major clinicopathological manifestations of CAA include cerebral hemorrhage, ischemic lesions, and dementia. CAA may be classified according to the amyloid protein deposited. In the most common form, sporadic CAA, and in CAA related to sporadic Alzheimer disease (AD). A beta deposition is characteristic. CAA can also be severe in variants of familial AD caused by mutations of the amyloid-beta precursor protein or presenilin-1 genes in which deposition of A beta variants and/or wild-type A beta occurs. Other amyloid proteins involved in familial CAAs include 1) the mutant cystatin C (ACys) in hereditary cerebral hemorrhage with amyloidosis of Icelandic type, 2) variant transthyretins (ATTR) in meningo-vascular amyloidoses, 3) mutated gelsolin (AGel) in familial amyloidosis of Finnish type, 4) disease-associated prion protein (PrP(Sc)) in a variant of the Gerstmann-Straussler-Scheinker syndrome, and 5) ABri and ADan in CAAs observed in the recently described BRI2 gene-related dementias, familial British dementia and familial Danish dementia, respectively. This review addresses issues related to the correlation between morphology, biochemistry, and genetics, and briefly discusses both the pathogenesis and animal models of CAAs. PMID- 14533780 TI - Decline in rate of colonization of oligodendrocyte progenitor cell (OPC)-depleted tissue by adult OPCs with age. AB - Rates of remyelination decline with age and this has been attributed to slower recruitment of oligodendrocyte progenitor cells (OPCs) into areas of demyelination and slower differentiation of OPCs into remyelinating oligodendrocytes. When considering causes for reduced recruitment rates, intrinsic causes (alterations in biological properties of OPCs) need to be separated from extrinsic causes (age-related differences in the lesion environment). Using 40 Gy of X-irradiation to deplete tissue of its endogenous OPC-population, we examined the effects of age on the rate at which adult rat OPCs colonize OPC-depleted tissue. We found a significant reduction in the rate of colonization between 2 and 10 months of age (0.6 mm/week versus 0.38 mm/week). To determine if this represented an intrinsic property of OPCs or was due to changes in the environment that the cells were recolonizing, OPCs from 10-month old animals were transplanted into 2-month-old hosts and OPCs from 2-month-old animals were transplanted into 10-month-old hosts. These experiments showed that the transplanted OPCs retained their age-related rate of colonization, indicating that the decline in colonizing rates of OPCs with age reflects an intrinsic property of OPCs. This age-related decline in the ability of OPCs to repopulate OPC-depleted tissue has implications for understanding remyelination failure in multiple sclerosis (MS) and developing therapies for remyelination failure. PMID- 14533779 TI - CX3CL1 and CX3CR1 expression in human brain tissue: noninflammatory control versus multiple sclerosis. AB - An important role for CX3CL1 in neuroinflammation and neurodegeneration has been suggested in recent publications. In this study, we compared the expression of CX3CL1 and its receptor CX3CR1 in human brain tissue derived from control patients without neurological complications and in multiple sclerosis (MS) patients. Results from this study demonstrate that CX3CL1 is constitutively expressed in human central nervous system (CNS) astrocytes in vivo and under basal conditions in human adult astrocyte cultures. CX3CR1 is expressed on astrocytes and microglial cells both in vivo and in vitro. Chemotaxis assay shows a functional response upon CX3CR1 signaling in microglial cells. Although CX3CL1 expression is upregulated in cultured astrocytes in response to proinflammatory cytokines, no evidence for expression differences of CX3CL1 between control patients and MS patients was found. Our data suggest that CX3CL1 has more general physiological functions, which occur also in the absence of proinflammatory conditions. PMID- 14533781 TI - Parallel compensatory and pathological events associated with tau pathology in middle aged individuals with Down syndrome. AB - Aged individuals with Down syndrome (DS) develop senile plaques and neurofibrillary tangles consistent with Alzheimer disease (AD). Prior to or in parallel with AD pathology, compensatory growth responses may occur. Immunohistochemistry and confocal microscopy studies in the hippocampus from 15 individuals ranging in age from 5 months to 67 years compared markers of normal and abnormal tau accumulation (phosphorylated tau [AT8, MC-1], tau-1, N-terminal tau) with the extent and location of neuronal growth marker immunoreactivity (BDNF, GAP-43, MAP-2). In middle age (30-40 years), prior to entorhinal neuron loss, the earliest tau accumulation occurred in the outer molecular layer (OML), which was consistent with both pathological and compensatory fetal tau expression. These events were followed at a later age, associated with entorhinal neuron loss, by an increase in GAP-43. Hilar neurons exhibiting a sprouting morphology were also noted. Age-dependent observations in the DS brain in the current study parallel hippocampal compensatory responses described in entorhinal cortex lesion studies in rodents. Thus, compensatory growth responses may occur in DS prior to extensive AD pathology and may be one mechanism underlying observations in PET studies of hypermetabolism in the entorhinal cortex of individuals with DS. PMID- 14533782 TI - Structural genomic abnormalities of chromosomes 9 and 18 in myxopapillary ependymomas. AB - Myxopapillary ependymomas (MPEs) are low-grade neuroepithelial tumors typically occurring in the conus-cauda equina-filum terminale region. Limited molecular and cytogenetic analysis of MPEs has not demonstrated consistent abnormalities. In an attempt to clarify the chromosomal status of these tumors and identify commonly aberrant regions in the genome we have combined 3 molecular/cyto/genetic methods to study 17 MPEs. Comparative genomic hybridization of 7/17 tumors identified concurrent gain on chromosomes 9 and 18 as the most frequent finding. The majority of the 17 tumors were also studied using microsatellite analysis with marker spanning the whole chromosomes 9 and 18 and interphase-FISH with centromeric probes for both chromosomes. Our combined results were consistent with concurrent gain in both chromosomes 9 and 18 in 11/17 cases, gain of either chromosome 9 or 18 and imbalance in the other chromosome in 3/17 tumors and allelic imbalances of chromosomes 9 or 18 in 3/17 and 1/17 tumors, respectively. Other abnormalities observed included gain of chromosomes 3, 4, 7, 8, 11, 13, 17q, 20, and X and loss of chromosomes 10 and 22. Our findings represent some steps towards understanding the molecular mechanisms involved in the development of MPE. PMID- 14533783 TI - Neurofilament RNA causes neurodegeneration with accumulation of ubiquitinated aggregates in cultured motor neurons. AB - The mechanisms whereby mutant gene expression triggers neurodegeneration are poorly understood but have generally been attributed to translated gene products. We now demonstrate direct neuropathic effects of untranslated RNA on cultured motor neurons. We show that expression of untranslated light neurofilament (NF-L) RNA sequence in the 3'UTR of an EGFP transgene (pEGFP/NF-L RNA) or in a separate expression vector (pRc/NF-L RNA) causes dose-dependent, neuron-specific motor neuron degeneration. Neither unfused EGFP protein (pEGFP/wt) nor EGFP-tagged NF-L protein (pEGFP/NF-L protein) has similar neuropathic effects. The findings are the first demonstration of a direct RNA-mediated neurotoxic effect. Moreover, the resulting neuropathological changes show that untranslated RNA can lead to early degeneration of neuritic processes and accumulations of ubiquitinated aggregates in the perikarya and nuclei of degenerating motor neurons. The latter findings are hallmark neuropathological features of neurodegenerative diseases and their occurrence as a result of altered RNA expression raises the prospects of an RNA mediated component in the pathogenesis of neurodegenerative states. PMID- 14533784 TI - Resetting the problem of cell death following muscle-derived cell transplantation: detection, dynamics and mechanisms. AB - We conducted a study in mice to reevaluate and clarify many aspects of the early survival of muscle cells following transplantation. Male mouse muscle cells (primary-cultures and T-antigen-immortalized clones) labeled with [14C]thymidine and beta-galactosidase were injected into female muscles. Each label was detected in the muscles after different time periods. TUNEL, alizarin red, and immunodetection of active caspase-3 were done in muscle sections. The donor cell labels disappeared from the muscles following donor cell death, but this was not instantaneous and even if the donor cells were killed before transplantation, the first 6 hours were not enough to clear [14C]thymidine and Y chromosome. Using the cell pellet before injection as the 100% baseline for cells injected to evaluate cell death can lead to misinterpretations: the Y-chromosome band was 5-fold stronger than that of a muscle injected with cells, irrespective of whether the cells were previously killed or not. There was no evidence of an immediate massive donor cell death. Necrosis (detected by alizarin red) and apoptosis (detected by active caspase-3) were present among the donor myoblasts following transplantation. Necrosis seemed to be the most important mechanism during the first hours. T-antigen immortalized cells died earlier and more massively than primary-cultured cells, but the surviving cells proliferated more. Indeed, they seemed to exhibit more apoptosis and they triggered a more rapid CD8+ cell infiltration. As a result of our findings, many concepts concerning the early donor cell death following myoblast transplantation must be reconsidered. PMID- 14533785 TI - Temporal course of upregulation of Na(v)1.8 in Purkinje neurons parallels the progression of clinical deficit in experimental allergic encephalomyelitis. AB - Multiple sclerosis (MS) is recognized to involve demyelination and axonal atrophy but accumulating evidence suggests that dysregulated sodium channel expression may also contribute to its pathophysiology. Recent studies have demonstrated that the expression of Na(v)1.8 voltage-gated sodium channels, which are normally undetectable within the CNS, is upregulated in cerebellar Purkinje cells in experimental allergic encephalomyelitis (EAE) and MS, and suggest that the aberrant expression of these channels contributes to clinical dysfunction by distorting the firing pattern of these neurons. In this study we examined the temporal pattern of upregulation for Na(v)1.8 mRNA and protein in chronic relapsing EAE by in situ hybridization and immunocytochemistry, respectively. Our results demonstrate a positive correlation between disease duration and degree of upregulation of Na(v)1.8 mRNA and protein in Purkinje neurons in chronic relapsing EAE. The progressive deterioration in clinical baseline scores (i.e. in clinical scores during remissions) is paralleled by a continued increase in Na(v)1.8 mRNA and protein expression, but temporary worsening during relapses is not associated with transient changes in Na(v)1.8 expression. These results provide evidence that the expression of sodium channel Na(v)1.8 contributes to the development of clinical deficits in an in vivo model of neuroinflammatory disease. PMID- 14533786 TI - The structural basis for carbohydrate recognition by lectins. AB - 1. Different carbohydrate-specific proteins, such as lectins, may combine with the same monosaccharide or oligosaccharide by different H-bonding and hydrophobic side chains. 2. Homologous proteins with distinct specificities may bind different monosaccharides (e.g., for glucose and galactose that differ in the configuration of a single hydroxyl) by the same set of invariant residues that are identically positioned in their tertiary structures. 3. The energetics of protein-carbohydrate interactions cannot be derived from structural information. 4. Nature solves in a variety of different ways the problem of constructing combining sites for carbohydrates, just as it provides diverse solutions for other functions of proteins. PMID- 14533787 TI - Lectin cross-linking interactions with multivalent carbohydrates. AB - The present findings provide a molecular basis for a new paradigm of specificity in multivalent carbohydrate-lectin interactions, namely the formation of type 2 homogeneous cross-linked lattices between multivalent carbohydrates and lectins. The present x-ray data demonstrate that the cross-linked complexes formed between a series of structurally related divalent carbohydrates and a single tetravalent lectin (SBA) are distinct and due to crystal packing interactions. These results thus provide a molecular basis for the formation of homogeneous type 2 cross linked complexes between lectins and multivalent carbohydrates and glycoconjugates. These findings are also relevant to the observations that lectin carbohydrate cross-linking interactions are involved in cellular recognition and signal transduction processes. For example, activated human T-cells undergo apoptosis due to binding and cross-linking of specific glycoprotein receptors by galectin-1 (Pace et al., 1999). Confocal microscopy shows that the galectin cross linked glycoprotein receptors form homogeneous aggregates from a population of previously dispersed molecules on the surface of the cells. The crystal structures of the four SBA/pentasaccharide complexes thus repesent models for lectin-carbohydrate clustering in vivo. PMID- 14533788 TI - Classification of plant lectins in families of structurally and evolutionary related proteins. AB - The majority of plant lectins can be classified in seven families of structurally and evolutionary related proteins. Within a given lectin family most but not necessarily all members are built up of protomers with a similar primary structure and overall 3-D fold. The overall structure of the native lectins is not only determined by the structure of the protomers but depends also on the degree of oligomerization and in some cases on the post-translational processing of the lectin precursors. In general, lectin families are fairly homogeneous for what concerns the overall specificity of the individual lectins, which illustrates that the 3-D structure of the binding site has been conserved during evolution. In the case of the jacalin-related lectins the occurrence of a mannose and galactose-binding subfamily can be explained by the fact that a post translational cleavage of the protomers (of the galactose-binding subfamily) yields a slightly altered binding site. Unlike the other families, the legume lectins display a wide range of specificites, which is clearly reflected in the occurrence of sugar-binding sites with a different 3-D structure. PMID- 14533789 TI - Expression of binding properties of Gal/GalNAc reactive lectins by mammalian glycotopes (an updated report). AB - Expression of the binding properties of Gal/GalNAc specific lectins, based on the affinity of decreasing order of mammalian glycotopes (determinants) rather than monosaccharide inhibition pattern, is probably one of the best ways to express carbohydrate specifity and should facilitate the selection of lectins as structural probes for studying mammalian glycobiology. Eleven mammalian structural units have been selected to express the binding domain of applied lectins. They are: 1. F, GalNAcalpha1 --> 3GalNAc; 2. A, GalNAcalpha1 --> 3Gal; 3. T, Galbeta1 --> 3GalNAc; 4. I, Galbeta 1 --> 3GlcNAc; 5. II, Galbeta1 --> 4GlcNAc; 6. B, Galalpha1 --> 3Gal; 7. E, Galalpha1--> 4Gal; 8. L, Galbeta1 --> 4Glc; 9. P, GalNAcbeta1 --> 3Gal; 10. S, GalNAcbeta1 --> 4Gal and 11. Tn, GalNAcalpha1 --> 4Ser (Thr) of the peptide chain. Thus, the carbohydrate specificity of Gal/GalNAc reactive lectins can be divided into classes according to their highest affinity for the above disaccharides and/or Tn residue. Examples of the binding properties of these lectins can be demonstrated by Ricimus communis agglutinin (RCA1), grouped as II specific lectin and its binding property is II > I > B > T; Ahrus precatorius agglutinin (APA), classified as T and its carbohydrate specificity is T > I/II > E > B > Tn; Artocarpus integrifolia (jacalin, AIL), as T/Tn specific and its binding reactivity is T > Tn >> I (II) and Geodia cydonium (GCL), as F/A specific, and with affinity for F > Ah [GalNAcalpha1-->43(L(Fuc)alpha1-->2)Gal] >> I > L. Due to the multiple reactivity of lectins toward mammalian glycotopes, the possible existence of different combining sites or subsites in the same molecule has to be examined, and the differential binding properties of these combining sites (if any) have to be characterized. To establish the relationship among the amino acid sequences of the combining sites of plant lectins and mammalian glycotopes should be an important direction to be addressed in lectinology. PMID- 14533791 TI - Animal lectins: from initial description to elaborated structural and functional classification. AB - The genetic code connects the two biochemical dimensions of nucleic acids and proteins. Theoretical calculations on coding capacity reveal that oligosaccharides as hardware surpass peptides by more than seven orders of magnitude based on hexamer synthesis. Thus, the sugar code establishes the third dimension of biological information transfer. Using carbohydrate-binding proteins (lectins, enzymes and antibodies) the information content of such epitopes is decoded. Currently, five families of animal lectins are defined in structural terms. i.e. the C-type, I-type and P-type groups, the galectins and the pentraxins. They are involved in intra- and intercellular glycan routing using oligosaccharides as postal-code equivalents and acting as defense molecules homing in on foreign or aberrant glycosignatures, as crosslinking agent in biosignaling and as coordinator of transient or firm cell-cell/cell-matrix contacts. By delineating the driving forces toward complex formation, knowledge about the causes for specificity can be turned into design of custom-made high affinity ligands for clinical application, e.g. in anti-adhesion therapy, drug targeting or diagnostic histopathology. PMID- 14533790 TI - 'Glyco-epitope' assignments for the selectins: advances enabled by the neoglycolipid (NGL) technology in conjunction with synthetic carbohydrate chemistry. AB - The neoglycolipid (NGL) technology involving the preparation of lipid-linked oligosaccharide probes for binding experiments with carbohydrate-recognizing proteins, and their analysis by mass spectrometry, is a unique and powerful means of discovering oligosaccharide ligands for carbohydrate-binding proteins, and assigning details of their specificities. The key feature is that it enables the pinpointing and sequence determination of bioactive oligosaccharides within highly heterogeneous mixtures derived from natural glycoconjugates. A new generation of NGLs incorporating a fluorescent label now establishes the principles for a streamlined technology whereby oligosaccharide populations are carried through ligand detection and isolation steps, and sequence determination. Advances in selectin research made through applications of the NGL technology include (i) demonstration of the importance of density of selectin expression, and of oligosaccharide ligands, in the magnitude and the specificity of the binding signals; (ii) demonstration of the efficacy of lipid-linked oligosaccharides in supporting selectin-mediated cell interactions; (iii) the discovery of 3-sulphated Le(a)/Le(x) as selectin ligands; (iv) the isolation and sequencing of carbohydrate ligands for E-selectin on murine myeloid cells and kidney; (v) the finding that sulphation at position 6 of the penultimate N acetylglucosamine confers superior L-selectin binding signals not only to 3 sialyl-Le(x) but also to 3'-sulpho-Le(x); and (vi) the finding that sialic acid de-N-acetylation, or further modification with formation of an intra-molecular amide bond in the carboxyl group, enhances or virtually abolishes, respectively, the potency of the 6'-sulfo-sialyl-Le(X) ligand. Working with biotinylated forms of the oligosaccharide ligands, we have observed that their presentation on a streptavidin matrix influences differentially the efficacy of interactions of the L- and P-selectins (but not E-selectin) with the sialylated and sulphated ligands. PMID- 14533792 TI - Galectins and other endogenous carbohydrate-binding proteins of animal bladder. AB - Defects in the glycocalyx of the bladder epithelium may be related to the development of bladder diseases including interstitial cystitis which is a chronic bladder disease of unknown etiology. Indirect evidence has implicated alterations in the bladder epithelial glycoconjugates in interstitial cystitis and vesicaler instillation of glycosaminoglycans is promoted as treatments. However, information on the nature of the glycoconjugates of the bladder epithelium and lectins that may interact with the exogenous instilled glycoconjugates is very limited. We have examined the endogenous lectin associated with bladder epithelium by immunohistochemistry using biotinylated neoglycoconjugates. The strong calcium-independent binding of beta-D-galactose probe suggested the presence of galectins in rabbit and human bladder. Extracts of rabbit bladder organ cultures metabolically labeled with [14C]-amino acids were subjected to affinity chromatography on immobilized lactose and the specifically bound material eluted with 0.2 M lactose. SDS-PAGE of the recovered proteins revealed a major band of approximately 30 kDa and a minor band of 21 kDa. Polymerase chain reaction and northern blot analysis showed that both galectin-3 and galectin-4 are expressed in rabbit bladder. Since galectin-3 from rabbit had been previously cloned, we cloned and sequenced galectin-4 from rabbit bladder. The deduced full length sequence of 328 amino acids revealed four distinct regions: a N-terminal peptide of 19 residues, two carbohydrate recognition domains of 130 residues each, and a linker region of 49 residues. Comparison of the rabbit galectin-4 sequence with those of human, pig, rat, and mouse revealed two invariant peptide motifs that are proposed as signature sequences for identifying related galectins. PMID- 14533793 TI - Binding properties and applications of Aplysia gonad lectin. AB - Adult Aplysia gonad contains high levels of a galactophilic lectin (MW around 65 kDa; composed of 2 subunits of apparent single species). It binds galactose and various alpha/beta-galactosides (but not N-acetylgalactosamine), in addition to an outstanding high affinity for galacturonic acid. This lectin is relatively resistant to heating up to 70 degrees C and to alkaline pH, but sensitive to proteolysis and low pH. It resembles galectins in binding to poly LacNAc (preferentially branched) complexes at low temperatures (0 degrees-4 degrees C) more avidly than at room temperature or at 37 degrees C, but differs from them in being Ca(2+)-dependent. It agglutinates papain/sialidase-treated erythrocytes more strongly than untreated cells and stimulates mitosis in peripheral human lymphocytes (inducing IL-2 formation). This lectin also enhances neurite outgrowth and increases their viability, while suppressing cell tumorigenicity. It is useful for histochemical/ cytochemical studies of galacturonic acid in plant tissues and fungi and for the study of cell surface composition of various prokaryotic (including halophilic Archaea) and eukaryotic cells and for their typing. It is useful as a reagent for I-antigen detection in adult human erythrocytes (anti-I), exhibiting strongest agglutination of O(h) Bombay-type erythrocytes and also exhibits sensitivity to the T antigen. It binds galactosylated molecules in human body fluids (shown by hemagglutination- inhibition tests), including saliva, seminal fluid and milk (detecting individual divergence) and in fowl egg albumens (exhibiting highest affinity for that of pigeon). Therefore, it might be valuable as a probe and fishhook for fishing compounds exhibiting anti-bacterial/neoplastic cell adhesion activities. PMID- 14533794 TI - Lectin and anti-carbohydrate antibody assays using chemically modified ligands. AB - Microtiter plate assays and 'lectinoblotting' with the use of biotinylated lectins are sensitive and easy to perform methods that can be combined with simple procedures of chemical modifications of glycoproteins immobilized on ELISA plates or blots (desialylation by mild acid hydrolysis, Smith degradation, beta elimination). These modifications are helpful in the determination of lectin and anti-carbohydrate antibody specificities, or in the characterization of glycoconjugates by means of lectins and antibodies. PMID- 14533795 TI - Intramolecular carbohydrate-protein interaction. AB - In this report, an overview is presented of studies on the three-dimensional structure in solution of glycoproteins. The investigations were carried out by using high resolution NMR spectroscopy in conjunction with molecular dynamics calculations and molecular modelling. For pine apple stem bromelain, ribonuclease 2 and the free alpha-subunit of human chorionic gonadotropin it could be shown that intramolecular interactions between carbohydrate and protein exist. Glycosylation sites are rather unique structural entities in glycoproteins, because of the local spatial arrangements. This is reflected in different intramolecular interactions between carbohydrate and protein. PMID- 14533797 TI - Antigenic properties of human glycophorins--an update. AB - Glycophorins are complex heavily glycosylated antigens carrying peptidic and glycopeptidic epitopes. Detailed immunochemical studies showed that GPA/GPB and GPC/GPD molecules have defined sites which are particularly immunogenic. These sites include N-terminal portions of all glycophorins, internal fragments of their extracellular domains, and cytoplasmic tails. The extracellular epitopes involve directly oligosaccharide chains (e.g. blood group M- and N-related epitopes, or N-terminal epitopes of GPC) or have peptidic character, shown by the reaction of respective antibodies with synthetic peptides. Peptidic eitopes are independent of glycosylation, or are variably affected by adjacent O-glycans which may mask the epitopes or may be required for a proper exposure of an antibody binding site. Several low incidence epitopes are present on variant glycophorin molecules. Among anti-glycophorin antibodies there are the 'bispecific' ones, or antibodies recognizing an epitope formed by an interaction of two proteins (Wr(b)). Alltogether, the glycophorins serve as convenient model antigens for studying Ag-Ab interaction and a role of O-glycosylation in protein antigenic properties. Moreover, well defined specificty of monoclonal anti glycophorin antibodies makes them more precise tools in serological investigation and identification of normal and variant antigens. Last but not least, elucidation of antigenic properties of glycophorins is important for identification and characterization of human anti-glycophorin antibodies, which in some cases create medical problems at transfusion or pregnancy. PMID- 14533796 TI - Red blood cell antigens responsible for inherited types of polyagglutination. AB - The three described types on inheritable polyagglutination are related to altered carbohydrate structures in glycoproteins or/and glycolipds on the erythrocyte surface. HEMPAS, a condition causing anemia and other pathological symptoms, is characterized by impaired biosynthesis of N-glycans, mostly those carried by band 3 and band 4.5 erythrocyte membrane proteins. Cad erythrocytes have abnormal glycophorin O-glycans, structurally related to the more common human Sd(a) and murine CT determinants, and accumulate an Sd(a)-like ganglioside. NOR erythrocytes express recently detected abnormal alpha-galactose-terminated glycosphingolipids, which strongly react with G. simplicifolia IB4 isolectin, but do not react with human anti-Galalpha1-3Gal antibodies. PMID- 14533799 TI - Glycoconjugates from parasitic helminths: structure diversity and immunobiological implications. AB - We have provided an account of the progress we and others have made over the last decade on the structural characterization of glycans from parasitic helminths. We hope to have illustrated a few principles and patterns governing helminth glycosylation, as well as the experimental approaches adopted and their associated strengths and limitations. Schistosomes remain the best studied systems but are still punctuated with gaps of knowledge. An important theme developed here is the regulated developmental stage-specific expression of various glycan epitopes and their interplay with immediate host environments for successful parasitism. It is anticipated that more novel or unusual structures will continuously be uncovered in the future and that despite many difficulties, current analytical techniques should be well up to meet the challenge in at least elucidating the major or key glycoconjugates from each of the diverse range of worms. The bottle neck will in fact reside in finding suitable experimental models to test their putative immunobiological functions from which the intricate host-parasite interactions can be delineated and rational vaccine design be achieved. The glycobiology of parasitic helminths is an area waiting to be more fully explored and the rewards should be sweet. PMID- 14533798 TI - KDN-containing glycoprotein from loach skin mucus. AB - It has been widely recognized that the mucus coat of fish plays a variety of important physical, chemical, and physiological functions. One of the major constituents of the mucus coat is mucus glycoprotein. We found that sialic acids in the skin mucus of the loach, Misgurnus anguillicaudatus, consisted predominantly of KDN. Subsequently, we isolated KDN-containing glycoprotein from loach skin mucus and characterized its chemical nature and structure. Loach mucus glycoprotein was purified from the Tris-HCl buffer extract of loach skin mucus by DEAE-cellulose chromatography, Nuclease P1 treatment, and Sepharose CL-6B gel filtration. The purified mucus glycoprotein was found to contain 38.5 KDN, 0.5% NeuAc, 25.0% GalNAc, 3.5% Gal, 0.5% GlcNAc and 28% amino acids. Exhaustive Actinase digestion of the glycoprotein yielded a glycopeptide with a higher sugar content and higher Thr and Ser contents. The molecular size of this glycopeptide was approximately 1/12 of the intact glycoprotein. These results suggest that approximately 11 highly glycosylated polypeptide units are linked in tandem through nonglycosylated peptides to form the glycoporotein molecule. The oligosaccharide alditols liberated from the loach mucus glycoprotein by alkaline borohydride treatment were separated by Sephadex G-25 gel filtration and HPLC. The purified sugar chains were analyzed b --> 6GalNAc-ol, KDNalpha2 --> 3(GalNAcbeta1 --> 14)GalNAc-ol, KDNalpha2 --> 6(GalNAcalpha1 --> 3)GalNAc-ol, KDNalpha2 --> 6(Gal3alpha1--> 3)GalNAc-ol, and NeuAcalpha2 --> 6Gal NAc-ol. It is estimated that one loach mucus glycoprotein molecule contains more than 500 KDN containing sugar chains that are linked to Thr and Ser residues of the protein core through GalNAc. PMID- 14533800 TI - Glycosyl phosphatidylinositol-linked glycoconjugates: structure, biosynthesis and function. AB - The purpose of this review is to summarize the most recent advances on GPI research. Structural studies on GPI-linked glycoconjugates indicate that there are significant variations in different organisms, although there is a conserved core structure. Furthermore, structural studies suggest that in different cell types, there is an army of glycosyltransferases dedicated to the synthesis of GPI linked glycoconjugates. Biochemical studies on the synthesis of these GPI-linked glycoconjugates suggest that not only many different enzymes are involved but also that enzymes from different cell types, involving in the conserved core structure can have different substrate specificity. Genetic cloning of the yeast genes involved in synthesizing the core structure suggests that many of these enzymes also have human homologues. However, paroxysmal nocturnal hemogobinuria (PNH) is the only known human disease associated with the synthesis of GPI-linked glycoconjugates. Functional studies suggest that GPI-anchor can act as a signal for protein sorting and localization. Furthermore, GPI-linked receptors play an important role in T-cell activation. PMID- 14533801 TI - Enzymatic and chemical approaches for the synthesis of sialyl glycoconjugates. AB - In conclusion, either enzymatic or chemical approaches have their unique features and unavoidable disadvantages. Enzyme-catalyzed sialylations provide the desired sialo-glycosidic linkages in the two enzyme reactions (CMP-NeuAc synthetase and sialyltransferase) with exclusive stereoselectivity and high yield as long as the required sialyltransferase is available. High substrate specificity of the two enzymes is a limitation so that many unnatural glycoconjugates cannot be prepared enzymatically. As for chemical glycosylations of sialic acids, it is possible to introduce any modification in sialyl donor and acceptor, in addition to create special sugar linkages. Nevertheless, reducing the number of reaction steps (for preparing both donors and acceptors of glycosylation), and enhancing stereoselectivity, as well as reaction yield are still problems to be overcome. PMID- 14533802 TI - The genetic regulation of fucosylated and sialylated antigens on developing myeloid cells. AB - The first part of this article reviews the stages of normal development of haemopoietic cells committed to the myeloid lineage, properties of leukaemic cell lines that are arrested at specific maturation stages along the granulocytic pathway, the structures of carbohydrate antigenic markers that appear on myeloid cell surfaces, with especial reference to sialyl-Le(x) (NeuAcalpha2-3Galbeta1 4[Fucalpha1-3]GlcNAc), and the role of this antigen on mature granulocytes as a ligand for selectin molecules. The families of fucosyl- and sialyltransferase genes encoding enzymes responsible for the biosynthesis of sialyl-Le(x), and the pathways leading to the formation of this antigen, and more complex related structures, are described. The second part of the article outlines the work carried out in the authors' laboratory with leukaemic cell lines in an attempt to ascertain the biochemical and genetic basis of the lowering of sialyl-Le(x) expression that occurs at intermediate stages of normal haemopoietic development. Analysis of enzyme levels and mRNA expression of the fucosyl- and sialyltransferase genes has led to the conclusion that depletion of substrate resulting from high levels of enzyme activity from co-expressed genes FUT4 and ST6Gal1 probably accounts for the dip in expression of sialyl-Le(x), rather than a change in the level of expression of FUT7, the gene in myeloid cells encoding the enzyme ultimately responsible for the synthesis of sialyl-Le(x). The possible significance of this change in relation to normal cell maturation is discussed. PMID- 14533803 TI - Transcriptional regulation of expression of carbohydrate ligands for cell adhesion molecules in the selectin family. AB - Cell adhesion mediated by selectins and their carbohydrate ligands is involved in the adhesion of cancer cells to endothelial cells during the course of hematogenous metastasis of cancer. In patients with leukemia, this adhesion is involved in the extravascular infiltration of leukemic cells. Extravasation and tissue infiltration of malignant cells in patients with adult T-cell leukemia is mediated by the interaction of selectins and their carbohydrate ligand sialyl Lewis X, which is strongly and constitutively expressed on the leukemic cells. Constitutive expression of Lewis X in these cells is due to the transcriptional activation of Fuc-T VII, the rate-limiting enzyme in the sialyl Lewis X synthesis, induced by the Tax protein encoded by the human T-cell leukemia virus 1, the etiological virus for this leukemia. This transactivation is in clear contrast to the regulation of typical CRE-element found in various cellular genes in that it is independent of phosphorylation-dependent regulation. This must be the reason for the strong and constitutive expression of sialyl Lewis X, which exacerbates the tissue infiltration of leukemic cells. This is a good example corroborating the proposition that the abnormal expression of carbohydrate determinant at the surface of malignant cells is intimately associated with the genetic mechanism of malignant transformation of cells. PMID- 14533804 TI - Biosynthesis and function of beta 1,6 branched mucin-type glycans. AB - The contribution of carbohydrate structure to biomolecular, cellular, and organismal function is well-established, but has not yet received the attention it deserves, perhaps due to the complexity of the structures involved and to a lack of simple experimental methods for relating structure and function. In particular, beta1,6 GlcNAc branching plays a key functional role in processes ranging from inflammation and immune system function to tumor cell metastasis. For instance, synthesis of the core 2 beta1,6 branched structure in the mucin glycan chain by C2GnT enables the expression of functional structures at the termini of polylactosamine chains, such as blood group antigens and sialyl Lewis x. Also, IGnT can create multiple branches on the polylactosamine chain, which may serve as a mechanism for amplifying the functional potency of cell surface glycoproteins and glycolipids. The family of enzymes which creates beta1,6 branched structure in mucin glycans is proving to be quite complex, since multiple isoforms appear to exist for these enzymes, and some of the enzymes are adept at forming more than one type of beta1,6 branched structure, as in the case of C2GnT-M. Furthermore, the enzymes do not appear to be restricted to acting on mucin-type acceptor structures, but are able to act on glycolipid structures as well. Much remains to be learned regarding the specific biological niche filled by each of these enzymes and how their activities complement one another, as well as the manner in which the activities of these enzymes are regulated in the cell. PMID- 14533805 TI - Differentiation-associated expression of beta-N-acetylgalactosaminylated N-linked oligosaccharides in mammary epithelial cells. AB - Not only mammalian pituitary glycoprotein hormones but also many glycoproteins from a variety of animal species have been shown to contain N-linked oligosacchardies with the GalNAcbeta1 --> 4GlcNAc structure. Two types of beta 1,4-GalNAcT were found; one transfers N-acetylgalactosamine to acceptor oligosaccharides, which is stimulated by the hormone peptide and the other simply transfers sugar without such activation. In the case of bovine mammary membrane glycoproteins, the expression of beta-N-acetylgalactosaminylated N-linked oligosaccharides was associated with the functional differentiation of the epithelial cells. In contrast, the expression level of such oligosaccharides was much reduced in glycoprotein samples from human breast tumors compared with those from the unaffected regions. These results strongly suggest that the beta-N acetylgalactosaminylation is one which is regulated under cellular differentiation and dedifferentiation of the mammary gland. Whether or not beta-N acetylgalactosaminylated N-linked oligosaccharides have unique functions in addition to clearance of the hormone from the circulation remains to be elucidated. PMID- 14533806 TI - Metabolism and role of O-acetylated sialic acids. PMID- 14533807 TI - Contradictory functions of the sulfatide in the blood coagulation system. AB - It has long been controversial whether sulfatide stimulates blood coagulation or inhibits blood coagulation. In this paper, I demonstrated not only anticoagulant activity but also coagulant activity of sulfatide in vivo by using experimental animal models and presented the possibility that sulfatide may function contradictorily under physiological and/or pathological conditions. For instance, I presented possible involvement of sulfatide in thrombotic diseases associated with cancers. PMID- 14533808 TI - Interaction of GM2 activator protein with glycosphingolipids. AB - GM2 activator protein is a protein cofactor that stimulates the hydrolysis of the GalNAc and the NeuAc in GM2 by beta-hexosaminidase A and sialidase, respectively. To understand the mechanism of action of GM2 activator, the interaction of this protein with GM2 and/or beta-hexosaminidase A has been the subject of interest since the purified GM2 activator became available. Numerous techniques including ultracentrifugation, isoelectric a focusing, polyacrylamide gel electrophoresis, gel filtration, thin layer chromatogram overlay, and fluorescence dequenching assay have been used to investigate the binding and the affinity of GM2 activator to various glycosphingolipids. It has been generally accepted that GM2 activator must have a very weak binding with the enzyme, because they can be easily separated from each other by gel filtration. Therefore, the interaction of GM2 and GM2 activator has been the focus for most of he study. Although preferential association of GM2 activator with GM2 was detected by some methods, GM2 activator was found also to bind other glycosphingolipids. Isolation of the specific complex that consists of only GM2 activator and GM2 from incubation mixture containing the activator protein and mixed glycosphingolipids has not been successfully carried out. Ultracentrifugation and gel-filtration are the mildest methods for the isolation of the complexes. However, these methods do not separate the complexes formed by specific interaction from that formed by non specific association. In fluorescence dequenching assay, the attempt to isolate the complex of R18 lipid probe with GM2 activator was also not successful. Since GM2 activator and glycosphingolipids contain hydrophobic domains in their molecules, the non-specific hydrophobic interactions between the two components can greatly interfere with the isolation of true functional complexes. Among the reported methods, thin layer chromatography overlay and the assay based on the inhibition of fluorescence dequenching by various glycosphingolipids are more informative than the others on the binding between GM2 activator and the carbohydrate head groups of glycosphingolipids. PMID- 14533809 TI - Tumor-associated carbohydrate antigens defining tumor malignancy: basis for development of anti-cancer vaccines. AB - Tumors expressing a high level of certain types of tumor-associated carbohydrate antigens (TACAs) exhibit greater metastasis and progression than those expressing low level of TACAs, as reflected in decreased patient survival rate. Well documented examples of such TACAs are: (i) H/Le(y)/Le(a) in primary non-small cell lung carcinoma; (ii) sialyl-Le(x) (SLe(x)) and sialyl-Le(a) (SLe(a)) in various types of cancer; (iii) Tn and sialyl-Tn in colorectal, lung, breast, and many other cancers; (iv) GM2, GD2, and GD3 gangliosides in neuroectodermal tumors (melanoma and neuroblastoma); (v) globo-H in breast, ovarian, and prostate cancer; (vi) disialylgalactosylgloboside in renal cell carcinoma. Some glycosylations and TACAs suppress invasiveness and metastatic potential. Well documented examples are: (i) blood group A antigen in primary lung carcinoma; (ii) bisecting beta1 --> 4GlcNAc of N-linked structure in melanoma and other cancers; (iii) galactosylgloboside (GalGb4) in seminoma. The biochemical mechanisms by which the above glycosylation changes promote or suppress tumor metastasis and invasion are mostly unknown. A few exceptional cases in which we have some knowledge are: (i) SLe(x) and SLe(a) function as E-selectin epitopes promoting tumor cell interaction with endothelial cells; (ii) some tumor cells interact through binding of TACA to specific proteins other than selectin, or to specific carbohydrate expressed on endothelial cells or other target cells (carbohydrate-carbohydrate interaction); (iii) functional modification of adhesive receptor (integrin, cadherin, CD44) by glycosylation. So far, a few successful cases of anti-cancer vaccine in clinical trials have been reported, employing TACAs whose expression enhances malignancy. Examples are STn for suppression of breast cancer, GM2 and GD3 for melanoma, and globo-H for prostate cancer. Vaccine development canbe extended using other TACAs, with the following criteria for success: (i) the antigen is expressed highly on tumor cells; (ii) high antibody production depending on two factors: (a) clustering of antigen used in vaccine; (b) choice of appropriate carrier protein or lipid; (iii) high T cell response depending on choice of appropriate carrier protein or lipid; (iv) expression of the same antigen in normal epithelial tissues (e.g., renal, intestinal, colorectal) may not pose a major obstacle, i.e., these tissues are not damaged during immune response. Idiotypic anti-carbohydrate antibodies that mimic the surface profile of carbohydrate antigens, when administered to patients, elicit anti-carbohydrate antibody response, thus providing an effect similar to that of TACAs for suppression of tumor progression. An extension of this idea is the use of peptide mimetics of TACAs, based on phage display random peptide library. Although examples are so far highly limited, use of such "mimotopes" as immunogens may overcome the weak immunogenicity of TACAs in general. PMID- 14533810 TI - Carbohydrate-mediated adhesion of human colon carcinoma cells to human liver sections. AB - Clinicopathological observations have revealed that the expression level of a carbohydrate antigen recognized by the monoclonal antibody (mAb) FH6 in colorectal carcinomas is higher at advanced stages than at early stages. The present study aimed to elucidate whether human colon carcinoma cell surface glycans recognized by mAb FH6 determine the ability of these cells to adhere to sections of human liver. Variant human colon carcinoma cell lines selected for high and low binding of mAb FH6 were compared with respect to their adhesive capacity. The cells expressing the higher level of FH6 binding also showed a greater ability to adhere to liver sections. This adhesion was not blocked by anti-E-selectin monoclonal antibodies, but pretreatment of the carcinoma cells with endo-beta-galactosidase significantly reduced both cell surface binding of mAb FH6 and the ability of the cells to adhere to liver sections. Our observations suggest that endo-beta-galactosidase-sensitive carbohydrate chains containing an epitope recognized by mAb FH6 play an important role in the adhesion of human colon carcinoma cells to human liver sections. Whether these interactions have any relationship to the mechanism(s) of liver metastasis remains to be elucidated. PMID- 14533811 TI - O-linked N-acetylglucosamine and cancer: messages from the glycosylation of c Myc. AB - Altered protein glycosylation is known to correlate with tumorigenesis, but its role remains enigmatic. Cells transformed by altered oncogene or tumor suppressor gene expression often also show changes of carbohydrate on cell surface glycoconjugates which correlate with the potential for tumor invasion and metastasis. In recent years, many oncogene and tumor suppressor gene products, such as c-Myc, SV40 large T antigen, and p53, were shown to be modified by O GlcNAc. O-GlcNAc is a form of protein glycosylation found almost exclusively in the nucleus and cytoplasm of eukaryotic cells. The known O-GlcNAc-bearing proteins are phosphoproteins and form reversible multimeric complexes. O-GlcNAc modification is dynamic and appears to have a reciprocal relationship with protein phosphorylation. The enzymes which catalyze O-GlcNAc addition and removal have been characterized and used as effective tools in O-GlcNAc studies. It is of great interest in the future to investigate the alteration of O-GlcNAc in different cancers since addition/removal of O-GlcNAc on oncoproteins, tumor suppressor proteins, and other tumor-related proteins very likely plays a key role in the pathogenesis of tumors. PMID- 14533812 TI - Analysis of a murine anti-ganglioside GD2 monoclonal antibody expressing both IgG2a and IgG3 isotypes: monoclonality, apoptosis triggering, and activation of cellular cytotoxicity on human melanoma cells. AB - In this study we have documented a hybridoma secreting an unusual MAb, which expresses both IgG3 and IgG2a subclasses with a lambda-light chain. How this dual expression of isotypes was exactly brought about is not clear. To resolve this problem, it will have to wait the complete sequence analysis the heavy chain gene of MAb 9C4. Although the expression of IgG2a was about 50% that of IgG3, antibody titration studies showed the major binding affinity of MAb 9C4 to GD3-positive cells being mostly contributed by the IgG3 rather than IgG2a part of the antibody. This antibody could induce apoptosis in melanoma cells in 10-15% of cells in vitro, but the generality of this phenomenon is yet to be confirmed by the use of different cell targets and different anti-GD2 MAbs other than 9C4. Aside from the demonstrated indirect killing mechanisms of many anti-GD2 MAbs through CDC and ADCC, MAb 9C4 induction of apoptosis represents an alternative mechanism of tumor cell killing, by which direct killing of anti-GD2 antibody takes its effect. This apoptotic effect was demonstrated for the first time with an anti-ganglioside monoclonal antibody. From the therapeutic point of view, the cytolytic activity of MAb 9C4-targeted ADCC/LAK killing against GD2-positive tumor cells to be more effective than that of LAK alone and a possibility for dendritic cells to effectively acquire antigen through pulsing with MAb-induced apoptotic cells are both of great clinical importance. Further studies are warranted aiming at elucidating the molecular basis of bi-isotypic specificity and aberrant isotype switching, molecular pathway of anti-GD2 antibody-induced apoptosis, and ways to improve clinical utility of this unusual hybridoma/MAb 9C4. PMID- 14533813 TI - Pathogen-host protein-carbohydrate interactions as the basis of important infections. AB - Microbe-host protein-carbohydrate interactions are most likely the essential first step to produce an infection, although this has been strictly proven only in a few cases. Improved glycotechnology will help identification of new carbohydrate receptors and this knowledge may be used to identify microbial carbohydrate-binding proteins by affinity proteomics approaches. In some cases such conserved proteins may prove to be successful vaccine components, in other cases, like influenza, saccharide analogues may be the only rational alternative. The prognosis may be, based on these improvements, that infection medicine will make considerable progress in the near future. PMID- 14533814 TI - Host mediated variation and receptor binding specificity of influenza viruses. AB - There are at least two possible mechanisms for the host range variation of influenza A viruses. First mechanism is the pressure of the antibody, this results has already reported by us, in 1989(12). In this case, Ser 205 which is located in antigenic site D in the hemagglutinin trimer. When Ser 205 is substituted to Tyr, receptor binding specificity become 2-6 predominant rather than 2-3 predominant. Ser205 is located far away from the receptor binding pocket in the subunit itself, however, very close to the pocket in the next subunit. When the Ser 205 is substituted to bulky amino acid Tyr, the three dimensional structure of the receptor binding pocket in next subunit may be influenced to become 2-6 binding property rather than 2-3 binding. Second mechanism is the selective pressure for the appearance of host cell variant with altered receptor binding specificities. In this case, amino acid 226 located in receptor binding pocket is clitical for the recognition of 2-6 and 2-3 linkages. Only single amino acid alteration of Leu 226 to Gln made remarkable change of receptor binding specificity 2-6 to 2-3. In this study, we also identified amino acid 155 Thr, 158 Glu, and 228 Ser of the hemagglutinin which may be clitical for the recognition of the molecular species of sialic acid, such as Neu5Ac and Neu5Gc. Very recently, we reported the amino acid residues contributing to the substrate specificity of the influenza A virus sialidase. If these amino acid alteration occurres in the host animal bodies in nature, the virus aquires the new receptor binding specificity which allows the infection to the different animal species, such as bird to human population. This process may be one of the important mechanism for the host mediated variation, and the influenza virus transmission between different hosts, and also emerging of new subtypes of influenza viruses in human population. PMID- 14533815 TI - Bacterial polysaccharides as vaccines--immunity and chemical characterization. AB - Studies on protective immunity and biochemical characterization of bacterial capsular polysaccharides have led to significant contributions to understanding of the mechanisms of infectious diseases and development of effective vaccines. Immunity to encapsulated bacteria is related to antibody response to polysaccharide (PS) antigen, interactions with T- and B-lymphocytes, and host defense mechanisms. Meningococcal, pneumococcal and Salmonella vi PSs and Haemophilus type b PS-protein conjugate vaccines have been licensed and provided effective immunity for prevention of these bacterial infections. Capsular PSs are cell-surface polymers consisting of oligosaccharide repeating units. Many PSs are highly polar and hydrophilic and interfere with cell-to-cell interactions with phagocytes. Most pneumococcal PSs are negatively charged and possess acidic components such as D-glucuronic acid and phosphate in phosphodiester bonds. Extensive immunologic cross-reactivity has been observed among bacterial capsular PSs. In infants the antibody responses to most capsular PSs are generally poor. Enhanced immunogenicity of PS antigens can be achieved through PS-protein conjugate vaccines, immunization during a critical period of perinatal development and effective antigen delivery system. PMID- 14533816 TI - The conformational epitope of type III group B Streptococcus capsular polysaccharide. AB - In order to further characterize the conformational epitope of GBSPIII, we synthesized various oligosaccharides with the GBSPIII-related structures by a tailor-assembly synthetic scheme and a more traditional block-wise chemo enzymatic approach. The oligosaccharides were used to probe the conformational epitope of GBSPIII using number of complementary techniques. The protective epitope of GPSPIII was further defined as length-dependent and conformational. The results of the studies confirmed that two repeating units (2RU) is the minimum binding unit and the epitope optimization mainly takes place between chain length 2RU to 7RU. Epitope optimization and multivalency were observed between 7RU and 20RU. The data support our hypothesis that the conformational epitope is an extended helical segment of the GBSPIII. GBSPIII exists mainly in the random coil form, which structurally mimics short oligosaccharide self antigens, but it can infrequently and spontaneously form extended helices. Although not prevalent in GBSPIII the immune system preferentially selects these helical epitopes because they are unique to the polysaccharide. PMID- 14533817 TI - T-independent IgA responses to microbial polysaccharides. AB - There is accumulating evidence indicating the presence in vivo of T-independent routes of IgA response in addition to the conventional T-dependent IgA response. Factors influencing these alternative pathways of IgA responses may include the structural characteristics of a stimulating antigen, the nature of responding B cells, and the microenvironment. The structural complexity of polysaccharide antigens has made it difficult to summarize a general scheme for the antibody responses they induce. Instead, one may expect that each individual polysaccharide may be able to create a unique microenvironment by activation of specific cell populations in the repertoires of non-T cell types. A specific pattern of B cell response may thus be elicited by TI stimulation. Recognition of such a unique property of a TI antigens is necessary for us to better understand the T-independent IgA response. Information obtained may have an impact on the development of vaccination strategies directed at the mucosal immunity mediated by IgA antibodies. PMID- 14533818 TI - Protective immunity and gene expression related to pneumococcal glycoconjugate. AB - The immunogenicity of most polysaccharides (PSs) contained in the pneumococcal vaccine is low in children less than 2 years of age. Enhancement of immune response in early life can be induced by immunization of pneumococcal glycoconjugate as well as plasma DNA coding for cell-surface protein antigen. Pneumococcal type 19F PS conjugated with inactivated pneumolysin (Ply) induced in mice remarkable antibody responses to both type 19F PS and the protein carrier. In addition, the conjugate was administered to pregnant mice during gestation and/or lactation, and to their offspring during early infancy. When the young mice were challenged with type 19F pneumococci, the bacteria were cleared more rapidly from the blood of immunized mice than from that of the control group. The mortality rate of young mice from immunized mothers was also significantly lower than the control group. These results indicate that the effective protective immunity against pneumococcal infection can be induced in young mice by the maternal immunization with the glycoconjugate during gestation and at early infancy. Studies have been conducted to express type 19A pneumolysin gene (ply) in mammalian cells. Ply DNA was inserted into the cloning site of a vector containing CMV promoter. The recombinant plasmid DNA containing ply was transfected in human rhabdomyosarcoma cells and the gene expression was confirmed by immunoblot. Injection of mice three times 50-100 ug per dose ply DNA produced high serum levels of Ply IgG and IgM antibodies and showed rapid bacterial clearance from the blood. PMID- 14533819 TI - Structure and functional genomics of lipopolysaccharide expression in Haemophilus influenzae. AB - The involvement of genes in the lic loci in H. influenzae LPS expression has been known for some time. However, it was not until recently that it was shown that the lic1 locus contains genes required for phase variable expression of phosphocholine substituents, while genes in the lic2 locus and lgtC are required for expression of the globoside trisaccharide, alpha-D-Galp-(1 --> 4)-beta-D-Galp (1 --> 4)-beta-D-Glcp (i.e., the pK blood group epitope). The availability of the complete sequence of the H. influenzae strain Rd genome has facilitated significant progress in understanding the role of these and other genes in the expression and biosynthesis of LPS. We have employed a comparative structural fingerprinting strategy to establish the structural relationships among LPS from H. influenzae mutant strains in which putative biosynthesis genes were inactivated. Using this functional genomics approach, we have gained considerable insight into the genetic basis for intra-strain and strain-to-strain variation in epitope expression. PMID- 14533820 TI - Molecular mimicry of host structures by lipooligosaccharides of Neisseria meningitidis: characterization of sialylated and nonsialylated lacto-N neotetraose (Galbeta1-4GlcNAcbeta1-3Galbeta1-4Glc) structures in lipooligosaccharides using monoclonal antibodies and specific lectins. AB - Neisseria meningitidis lipooligosaccharides (LOSs) are classified into 12 immunotypes. Most LOSs are heterogeneous in having a few components by SDS-PAGE analysis that differ antigenically and chemically. We have utilized a monoclonal antibody that recognizes lacto-N-neotetraose (LNnT) and the lectin, Maackia amurensis leukoagglutinin (MAL), which is specific for NeuNAcalpha2-3Galbeta1 4GlcNAc trisacchride sequence to characterize the 12 N. meningitidis LOSs. Using the combination of ELISA, SDS-PAGE, Western blotting, and other chemical analyses, we have shown that the LNnT (Galbeta1-4GlcNAcbeta1-3Galbeta1-4Glc) sequence was present in the 4.0-kDa LOS components of seven immunotype LOSs seen on SDS-PAGE. Six of the seven LNnT-containing LOSs also bound the MAL lectin indicating that N-acetylneuraminic acid (NeuNAc) was alpha2,3-linked to the LNnT sequence in the LOSs. Sialylation of the terminal Gal of LNnT-containing 4.0-kDa component caused only a slight increase in its apparent MW to 4100 on SDS-PAGE. The one LOS with the LNnT-containing component, but not MAL-binding, was from a Group A N. meningitidis, which does not synthesize CMP-NeuNAc, the substrate needed for LOS sialylation. Thus, it is concluded (1) a common LNnT sequence is present in seven immunotype LOSs in addition to their immunotype epitopes, and (2) NeuNAc is alpha2 --> 3 linked to the terminal Gal of LNnT if a organism synthesizes CMP-NeuNAc such as Groups B and C organisms. The above conclusions are consistent with the published structures of N. meningitidis LOSs. The results also demonstrate that specific carbohydrate-binding lectins and monoclonal antibodies can be used as simple yet effective tools to characterize specific carbohydrate sequences in a bacterial LOS or LPS such as N. meningitidis LOS. It is intriguing that N. meningitidis LOSs mimic certain glycosphingolipids, such as paragloboside (LNnT-ceramide) and sialylparagloboside, and some glycoproteins of the host in having LNnT and N-acetyllactosamine sequences respectively with or without alpha2 --> 3 linked NeuNAc. Epidemiological studies of N. meningitidis suggest that the molecular mimicry of host structures by its LOS plays a role in the pathogenesis of N. meningitidis by helping the organism to evade host immune defenses in man. The molecular mimicry of host structures by LOS or LPS is also found in other human pathogens such as N. gonorrhoeae, Haemophilus ducreyi, H. influenaze, Moraxella catarrhalis, Campylobacter jejuni, and Helicobacter pylori. PMID- 14533821 TI - NMR and molecular modeling of complex carbohydrates and carbohydrate-protein interactions. Applications to anti-bacteria vaccines. AB - In order to characterize the conformational epitope of the group B meningococcal polysaccharide and of the type III group B Streptococcus capsular polysaccharide NMR measurements were done on a wide variety of native and modified polysaccharides and oligosaccharides. Since these saccharides are highly mobile and exist as random coils in solution, the analysis of the NMR data and molecular modeling was done to take into account this inherent flexibility. The conformational model of extended high-order helices being selected upon binding to a protein, although still hypothetical at this stage, has proven useful in explaining the serology for the conformational epitopes for polysaccharides of group B Neisseria meningitidis, group B Streptococcus type III and Streptococcus pneumoniae type 14. PMID- 14533822 TI - A guide to the carbohydrate specificities of applied lectins-2 (updated in 2000). PMID- 14533823 TI - A guide to monoclonal antibodies directed to glycotopes. PMID- 14533825 TI - Implications of UV-induced inflammation and immunomodulation. AB - Sunscreens are the most effective and widely available interventions for sun damage, other than sun avoidance or clothing. However, sun-screens vary widely in their ability to screen various UV wavelength components. Testing methods for sunscreens rely on UV-induced erythema to determine a sun protection factor (SPF), primarily a measure of UVB protection only. Determination of an immune protection factor (IPF) has been proposed as an alternative or adjunctive measure to SPF, and, indeed, recent studies show that the IPF can detect the added in vivo functionality of sunscreens--such as high levels of UVA protection--that the SPF cannot. Consensus on the definition of IPF, however, is required. Data are available on quantification of the IPF for restoring the afferent or induction arm of contact sensitivity, but other immune parameters also have been measured. A review of in vivo studies in humans, in which sunscreens are used to intervene in UV-induced modulation of immune response, cells, or cytokines, highlights the technical variables and statistical approaches that must be standardized in the context of an IPF for regulatory product claim purposes. Development of such IPF standards would allow the integration of both UVB and non-UVB solar wave-band effect-reversals. In addition, it could be applied to integrate the effects of other ingredients with protective function (ie, antioxidants, retinoids, or other novel products) and spur the development of more advanced and complete protection products. PMID- 14533824 TI - Skin aging 2003: recent advances and current concepts. AB - In developed countries, interest in cutaneous aging is in large part the result of a progressive, dramatic rise over the past century in the absolute number and proportion of the population who are elderly. The psychosocial, as well as physiologic, effects of skin aging on older persons have created a demand for a better understanding of the aging process and particularly for effective interventions. Skin aging is a complex process determined by the genetic endowment of the individual and by environmental factors. The appearance of old skin and the clinical consequences of skin aging have been well-known for centuries, but it is only in the past 50 years that mechanisms and mediators have been pursued systematically. Still, within a relatively short time, there has been tremendous progress, a progress greatly enhanced by basic gerontologic research using immunologic, biochemical, and in particular, molecular biologic approaches. PMID- 14533826 TI - Medical implications of DNA repair. AB - When the sun damages the skin, it also impairs the DNA. The DNA repair system is needed to maintain the genetic integrity of the epidermis. Defects in DNA repair (eg, xeroderma pigmentosum) commonly result in skin cancer. Sun-screens are important for preventing sun damage, but inadequate application by consumers is common, and protection against DNA damage is incomplete. The human population varies widely in its formation and response to DNA damage. Sequencing of individuals' DNA repair genes has revealed unexpected diversity, and some polymorphisms may be related to skin cancer risk. DNA damage has been linked to immune suppression in humans, and variations in this immune response also are linked to cancer risk. PMID- 14533827 TI - Problem pets. PMID- 14533828 TI - Botanical briefs: the century plant--Agave americana L. PMID- 14533829 TI - Trichotillomania: a review and case report. AB - Although patients with trichotillomania typically present to dermatologists, the diagnosis and treatment lie in the field of psychiatry. We report an unusual case of a 33-year-old woman with severe trichotillomania. We review common clinical and pathologic findings of this often chronic and socially debilitating disorder. In addition, we discuss treatment options for dermatologists and how collaboration with psychiatrists is the most effective management for these difficult-to-treat patients. PMID- 14533830 TI - What is your diagnosis? Nevus lipomatosus cutaneous superficialis. PMID- 14533831 TI - Childhood cutaneous hemangiomas. AB - An hemangioma is a common benign growth of vascular endothelium that may have multiple clinical manifestations. Either identified at birth or shortly thereafter, hemangiomas have both proliferative and involutional phases. Certain varieties are associated with congenital anomalies. Numerous therapy options are discussed. PMID- 14533832 TI - Wells syndrome. AB - Wells syndrome, or eosinophilic cellulitis, is a rare disorder characterized by erythematous plaques evolving into dermal or subcutaneous masses. The histopathology shows degenerative collagen, histiocytes, and eosinophils. Although most cases of Wells syndrome occur in adults, we present a child with these clinical and histopathologic findings. PMID- 14533833 TI - Granuloma faciale: distribution of the lesions and review of the literature. AB - Granuloma faciale (GF) is an uncommon inflammatory condition characterized by reddish brown papules and plaques that usually involve the facial area. Extrafacial lesions are rare. Histologically, the lesions are marked by leukocytoclastic vasculitis and extensive fibrin deposition. There are a variety of treatment options available for GF. PMID- 14533834 TI - Widespread cutaneous involvement by invasive Apophysomyces elegans in a gravid patient following trauma. AB - Invasive infections in humans with organisms from the fungal subclass Zygomycetes are most commonly seen in immunocompromised and diabetic patients. Rarely, such fungal infections may be seen in immunocompetent, nondiabetic individuals. In these cases, cutaneous trauma with direct implantation of fungal organisms into the wound from soil contamination is the frequent scenario. We present the case of a 31-year-old gravid woman involved in a single-vehicle automobile accident who presented to our institution with severe head trauma. On admission, a small ecchymotic area on her right forearm was noted. The lesion eventually expanded and ulcerated. Culture and histologic examination of tissue from the site revealed fungal organisms consistent with Zygomycetes. Subsequent studies confirmed the fungal organism as Apophysomyces elegans. Antifungal therapy was initiated, and multiple debridements were performed. Amputation of the right arm above the elbow was eventually necessary, but aggressive surgical intervention and antifungal therapy were unsuccessful in preventing the spread of the infection. The patient died 2 weeks after admission from polymicrobial sepsis. This case illustrates the dangerously invasive nature of A elegans, even in immunocompetent individuals. PMID- 14533835 TI - Marked hyperkeratosis of the soles in keratitis-ichthyosis-deafness syndrome: treatment with hydrocolloid dressing. AB - Keratitis-ichthyosis-deafness (KID) syndrome is a rare congenital disorder that exhibits marked hyperkeratosis of the skin. We successfully treated cutaneous lesions on the soles of a patient with KID syndrome using hydrocolloid dressing. PMID- 14533836 TI - Onycholysis with the appearance of a "sunset" secondary to capecitabine. AB - Capecitabine is an antineoplastic agent that is currently the only effective treatment for patients with advanced or metastatic breast cancer in whom anthracycline or taxoid treatment has failed. Dermatologic side effects have included hand-foot syndrome and one case of onycholysis. We report a case of onycholysis with appearance of a "sunset" induced by capecitabine. PMID- 14533837 TI - Comparison of azithromycin and cefadroxil for the treatment of uncomplicated skin and skin structure infections. AB - In this multicenter, investigator-blind trial, we compared the efficacy and safety of azithromycin and cefadroxil for the treatment of uncomplicated skin and skin structure infections (SSSIs). A total of 296 patients were randomized to receive either azithromycin (500 mg on day 1, followed by 250 mg once a day on days 2 to 5) or cefadroxil (500 mg twice a day for 10 days). Outpatients, ranging in age from 18 to 75 years, with acute uncomplicated SSSIs were enrolled in the study. Clinical and bacteriologic response was assessed between days 10 and 13 (primary end point) and between days 28 and 32. In a modified intent-to-treat analysis, clinical success rates assessed between days 10 and 13 were 97% (111/114) for azithromycin and 96% (101/105) for cefadroxil (P = .717). For azithromycin and cefadroxil, corresponding rates of bacteriologic eradication for Staphylococcus aureus were 94% (64/68) and 86% (60/70), respectively, and for Streptococcus pyogenes, 80% (4/5) and 100% (6/6), respectively. Clinical success rates assessed between days 28 and 32 were 100% (82/82) for azithromycin compared with 90% (75/83) for cefadroxil (P = .007). Corresponding rates of eradication for S aureus were 100% (59/59) versus 89% (56/63), respectively; and for S pyogenes, 100% (4/4) versus 83% (5/6), respectively. The incidence of treatment related adverse events was similar in the 2 treatment groups. However, 5 of the 139 patients (4%) in the cefadroxil group discontinued therapy because of treatment-related adverse events compared with none of the 152 patients in the azithromycin group (P = .02). Five-day therapy with azithromycin was as effective as 10-day therapy with cefadroxil for treating uncomplicated SSSIs. PMID- 14533838 TI - Optimizing cochlear implant speech performance. AB - Results of studies performed in our laboratory suggest that cochlear implant recipients understand speech best if the following speech processor parameters are individually chosen for each person: minimum and maximum stimulation levels on each electrode in the speech processor program (MAP), stimulation rate, and speech coding strategy. If these and related parameters are chosen to make soft sounds (from approximately 100 to 6,000 Hz) audible at as close to 20 dB hearing level as possible and loud sounds not too loud, recipients have the opportunity to hear speech in everyday life situations that are of key importance to children who are learning language and to all recipients in terms of ease of communication. PMID- 14533839 TI - Signal coding in cochlear implants: exploiting stochastic effects of electrical stimulation. AB - Speech perception in quiet with cochlear implants has increased substantially over the past 17 years. If current trends continue, average monosyllabic word scores will be nearly 80% by 2010. These improvements are due to enhancements in speech processing strategies, to the implantation of patients with more residual hearing and shorter durations of deafness, and to unknown causes. Despite these improvements, speech perception in noise and music perception are still poor in most implant patients. These deficits may be partly due to poor representation of temporal fine structure by current speech processing strategies. It may be possible to improve both this representation and the dynamic range of electrical stimulation through the exploitation of stochastic effects produced by high-rate (eg, 5-kilopulse-per-second) pulse trains. Both the loudness growth and the dynamic range of low-frequency sinusoids have been enhanced via this technique. A laboratory speech processor using this strategy is under development. Although the clinical programming for such an algorithm is likely to be complex, some guidelines for the psychophysical and electrophysiological techniques necessary can be described now. PMID- 14533840 TI - Prosthetic stimulation of the auditory system with intraneural electrodes. AB - Prosthetic electrical stimulation of the auditory system is presently accomplished either via scala tympani electrode arrays or via cochlear nucleus surface electrode arrays. Many of the early cochlear implant studies, however, used electrode arrays placed within the auditory nerve itself--either within the modiolus or within the trunk of the nerve. For many reasons, such intraneural electrode arrays were abandoned in favor of intrascalar arrays. There remain, however, several theoretical and practical reasons why intraneural arrays might be advantageous, and recent developments in electrode technology solve many of the problems posed by early attempts at intraneural stimulation. In this article, we review the history and current status of intraneural auditory stimulation, and present some preliminary results of this mode of stimulation in an animal model. PMID- 14533841 TI - Technology and the future of cochlear implants. AB - Cochlear implants are one of the dramatic success stories of the bioengineering enterprise. Although these prostheses are used extensively, they still can be improved substantially. We suggest that high-density electrode designs will permit field shaping and field steering to an extent not presently possible with the arrays that are used today. Those opportunities will make it possible to make use of the phase information that is richly available to normal listeners. Although this information makes possible more precise location of sound sources in the auditory environment, and will likely improve the recognition of intelligent sound in noise, it will require the consumption of additional power in future cochlear prostheses. Those opportunities and trade-offs provide the designers of cochlear implants with exciting goals for the future. PMID- 14533842 TI - Identification and culture of olfactory neural progenitors from GFP mice. AB - The olfactory epithelium (OE) is one of the best sources for obtaining adult stem cells from the nervous system, because it contains neural progenitors that regenerate continuously throughout life. The OE is accessible through the nasal cavity, which facilitates stem cell harvest for examination and transplantation. The mitotic activity of OE progenitors can be stimulated by intranasal irrigation with zinc sulfate (ZnSO4). In the study reported here, we focused on OE from a transgenic mouse line transfected with green fluorescent protein (GFP). Histological examination demonstrated the site of highest yield of OE in the transgenic and wild type littermates. Cultures were established from that site four days in vitro following ZnSO4 exposure. The GFP-derived primary cultures contained a heterogeneous population of fluorescent cells. After 10-12 days, a population of round, mitotically active cells emerged that formed fluorescent neurospheres. The neurosphere forming cells (NSFCs) were collected and subcultured up to four times. The NSFCs were primarily neuronal with only a few cells of glial lineage. Furthermore, the NSFCs were nestin positive and keratin negative, suggesting that they were neural progenitors. The endogenous GFP fluorescence of these cells provides a readily identifiable label that will facilitate their identification following transplantation into nontransfected hosts. They should provide a useful model for evaluating the potential therapeutic utility of OE progenitors in neurodegenerative diseases and neurotrauma repair. PMID- 14533843 TI - Sieve element specific labeling with an anti-idiotypic monoclonal antibody mimic of the phytotoxin dothistromin. AB - A monoclonal antibody, 12C9, an anti-idiotypic mimic of dothistromin, a toxin produced by Dothistroma pini, was found to label the cell wall of sieve elements in a number of different plant tissues and species. The antibody labeled apple leaf tissue, tobacco leaf mid vein, leaf and meristem, and Coprosma robusta leaf mid vein. Labeling was restricted to cell walls of sieve elements and did not label the companion cells or the lumen of the cells. The antibody labeled over a wide range of dilutions. This antibody could be used to differentiate sieve elements from other types of phloem. It could also be used to co-localize sieve elements and microorganisms such as phytoplasmas stained with DAPI. PMID- 14533844 TI - Formats of image data files that can be used for routine digital light micrography. Part one. AB - Failing to open computer files that describe image data is not the most frustrating experience that the user of a computer can suffer, but it is high on list of possible aggravations. To ameliorate this, the structure of uncompressed image data files is described here. The various ways in which information that describes a picture can be recorded are related, and a primary distinction between raster or bitmap based and vector or object based image data files is drawn. Bitmap based image data files are the more useful of the two formats for recording complicated images such as digital light micrographs, whereas object based files are better for recording illustrations and cartoons. Computer software for opening a very large variety of different formats of digital image data is recommended, and if these fail, ways are described for opening bitmap based digital image data files whose format is unknown. PMID- 14533845 TI - Formats of image data files that can be used in routine digital light micrography. Part two. AB - This is part two of an article that describes the properties of the image data files that are encountered routinely in digital light micrography. In the current part of the article, the differences between saving image data as large intact files and smaller files that have had some information removed, i.e., using lossy compression, are related first. Subsequently, appropriate ways of configuring computers to deal with the large intact image data files are suggested. The structures of the image data files used for recording dynamic sequences and kinematic animations of series of digital light micrographs, i.e., movie formats, are then described. Finally, some information is supplied about choosing file formats for compressing both static and dynamic image data sets. PMID- 14533846 TI - Decreased activity of cathepsins L + B and decreased invasive ability of PC3 prostate cancer cells. AB - Cancer metastasis involves multiple factors, one of which is the production and secretion of matrix degrading proteases by the cancer cells. Many metastasizing cancer cells secrete the lysosomal proteases, cathepsins L and B, which implicates them in the metastatic process. Cathepsins L and B are regulated by endogenous cysteine proteinase inhibitors (CPI) known as cystatins. An imbalance between cathepsin L and/or B and cystatin expression/activity may be a characteristic of the metastatic phenotype. To determine whether cystatins can attenuate the invasive ability of PC3 prostate cancer cells, cells were transfected with a cDNA coding for chicken cystatin. Expression of chicken cystatin mRNA was determined by PCR analysis. Total cysteine proteinase inhibitory activity, cathepsins L + B activity, and invasion through a Matrigel matrix were assessed. Stably transfected cells expressed the chicken cystatin mRNA and exhibited a significant decrease in secreted cathepsin L + B activity and a small increase in secreted cysteine proteinase inhibitor activity. The ability of cystatin transfected cells to invade the reconstituted basement membrane, Matrigel, was attenuated compared to nontransfected cells or cells transfected with vector alone. We have demonstrated that the cysteine proteinases cathepsins L and B participate in the invasive ability of the PC3 prostate cancer cell line, and we discuss here the potential of using cysteine proteinase inhibitors such as the cystatins as anti-metastatic agents. PMID- 14533847 TI - Cell shape and gene expression in human intervertebral disc cells: in vitro tissue engineering studies. AB - The objective of the present study was to examine the relation between gene expression and the shape of human intervertebral disc cells cultured in vitro in three-dimensional (3D) scaffolds. Disc cells from 19 subjects were seeded into either a collagen sponge or collagen gel and cultured for 10 days. In situ hybridization was performed on serial sections of paraffin embedded specimens and assessed for expression of selected genes important for extracellular matrix formation: Types I and II collagen, aggrecan and chondroitin-6 sulfotransferase. Rounded cells grown in collagen gel showed expression of Types I and II collagen, aggrecan and chondroitin-6 sulfotransferase; expression of these genes was absent in spindle shaped cells. Cells in the collagen sponge that lay on the sponge margin were frequently spindle shaped; these cells expressed type I collagen, but not type II collagen, aggrecan or chondroitin-6 sulfotransferase. Results presented here provide novel data concerning disc cell gene expression with collagen 3D constructs. This information is useful for future tissue engineering studies that have the challenging goal of selectively modulating gene expression. PMID- 14533848 TI - A comparison of microwave heating and proteolytic pretreatment antigen retrieval techniques in formalin fixed, paraffin embedded tissues. AB - Antigen retrieval (AR) is a technique that re-exposes epitopes in formalin fixed, paraffin embedded sections and makes them detectable by immunohistochemistry. We compared the effects of two AR procedures, enzyme digestion and microwave heating, on immunostaining of vimentin and desmin in formalin fixed, paraffin embedded tissues. Our results showed that AR is necessary for vimentin and desmin immunostaining in tissues fixed in formalin for more than 48 h. With prolonged fixation times, microwave heating showed better results than enzyme digestion for AR. The same results were obtained using 1% zinc sulfate or Citra Plus solution as retrieval solutions for microwave heating. We recommend microwave heating for AR, because it is easier to use and produces better results compared to enzyme treatment. PMID- 14533849 TI - A morphological and immunolabeling study of freeze-substituted Bacteroides forsythus. AB - We used a rapid freezing and freeze-substitution technique without glutaraldehyde and OsO4 fixation for the electron microscopic immunocytochemical demonstration of the surface structure of Bacteroides forsythus, an anaerobic Gram-negative periodontopathogen. Cells were applied to pieces of filter paper and freeze substituted by plunge-freezing in liquid propane, substituted in methanol containing 0.5% uranyl acetate, and infiltrated with LR White resin. The membrane ultrastructure of B. forsythus was preserved well, and the labeling density of the freeze-submitted cells was compared to a conventional processing method. Our results show the usefulness of the freeze-substitution method for immunohistochemical studies of B. forsythus. PMID- 14533850 TI - Stability and solubility of 3,3'-diaminobenzidine (DAB). PMID- 14533851 TI - Storage of frozen tissue blocks. PMID- 14533852 TI - Income, age and financial satisfaction. AB - Although the effects of income and age on subjective well-being have been widely studied, research on the effects of income and age on financial satisfaction, a major life domain to which income has direct relevance, remains limited. Analyzing data from the General Social Surveys, this article empirically examined the effects of income and age on financial satisfaction. These findings suggest that the social-psychological mechanisms underlying the age differences in the effects of income on financial satisfaction might not reflect a clear-cut status attainment versus status maintenance framework. The findings also served to caution future financial satisfaction research in the choice of income measures and the age grouping. PMID- 14533853 TI - Gender differences in physical health and psychosocial well being among four age groups of elderly people in Israel. AB - The purpose of this study was to investigate the extent to which the well established gender differences in physical and psychosocial well being in adulthood persist throughout different age groups of elderly persons, in order to support one of two opposing hypotheses: the convergence and divergence hypotheses. Data were collected by structured interviews from a random sample of 987 Israeli elderly (70+) in 1994. They were divided into four age groups for analysis: 70-74, 75-79, 80-84, and 85 and over. Findings indicate that in nearly every gender comparison by age, women score lower than men on indicators of physical and psychosocial well being, and in both genders increasing limitations on activities of daily living (ADL) were noticed. However, on all measures of physical health, except for ADL, the male advantage declines in the older age groups. This pattern is even stronger for the psychosocial indicators of well being, where no significant gender differences are found between the oldest groups. The trend of convergence among men and women thus occurs mainly in the age group of 85+. The results of multivariate analyses indicate that the sense of control of one's life is an important explanatory variable of satisfaction with life for men but not for women. The significant decline in the sense of control of men, aged 85 and over, is one of the main reasons for the more significant decline found in men's psychosocial well being in comparison to women. Our findings indicate that decline in health status, and other losses experienced with aging, affect more significantly men's sense of control over life, and therefore have a more deleterious effect on the psychosocial well being of men than on that of women. This conclusion, however, has to be supported by longitudinal studies. PMID- 14533854 TI - Attitudes toward embodied old age among Swedes. AB - Messages in the consumer culture are often youth oriented, aiming at the prevention of the bodily decay associated with biological aging. In gerontological discourses, this has been hypothesized to generate negative attitudes toward embodied aging and old age. Studies about general attitudes toward old age show that younger respondents have more negative attitudes than do older respondents, and gerontological discourses also hypothesize a gendered ageism, with especially negative attitudes toward elderly women. The empirical study of embodied aging among 1,250 Swedes aged 20-85 years contradicts these hypotheses. The results show rather positive attitudes toward embodied old age, especially among young and middle-aged respondents. Neither do the results unequivocally confirm the hypothesis of gendered ageism, which predicts considerably more negative attitudes toward old women than toward old men. One interpretation of the results is that, counter to many hypotheses, the consumer culture, with its new opportunities and roles for old people, may positively affect these attitudes. PMID- 14533855 TI - Preferences for prolonging life: a prospect theory approach. AB - Kahneman and Tversky's (1979) Prospect theory was tested as a model of preferences for prolonging life under various hypothetical health statuses. A sample of 384 elderly people living in congregate housing (263 healthy, 131 frail) indicated how long (if at all) they would want to live under each of nine hypothetical health conditions (e.g., limited to bed or chair in a nursing home). Prospect theory, a decision model which takes into account the individual's point of reference, would predict that frail people would view prospective poorer health conditions as more tolerable and express preferences to live longer in worse health than would currently healthy people. In separate analyses of covariance, we evaluated preferences for continued life under four conditions of functional ability, four conditions of cognitive impairment, and three pain conditions--each as a function of participant's current health status (frail vs. healthy). The predicted interaction between frailty and declining prospective health status was obtained. Frail participants expressed preferences for longer life under more compromised health conditions than did healthy participants. The results imply that such preferences are malleable, changing as health deteriorates. They also help explain disparities between proxy decision-makers' and patients' own preferences as expressed in advance directives. PMID- 14533856 TI - Material-dependent levels of heat-shock protein 70 (hsp70) in human plasma following contact of blood with artificial surfaces. AB - Recently, it has been shown that heat-shock protein 70 (hsp70) functions in a dual role as a chaperone and a cytokine. However, no information is available on the occurrence of hsp70 in the extracellular milieu or on its ability to modulate cellular immune response. This study shows a material-dependent increase of hsp70 levels in plasma following contact of fresh heparinized whole human blood with three different biomaterials (PVC, heparin-coated PVC, Silicone). We report a previously unknown behavior of hsp70 to act as a plasma-adsorption protein. Further, high binding capacities for hsp70 to artificial surfaces (measured by Western blotting) and elevated hsp70 levels in plasma (measured by EIA) following contact with blood correspond with a reduced hemocompatibility. The degree of surface-induced activation of blood was determined by analysis of markers for coagulation, inflammation and complement activation. These findings indicate that the selective adsorption of hsp70 on artificial surfaces and the increased hsp70 levels in plasma may be important in directing host inflammatory and immune responses. We suggest that the levels of hsp70 in human plasma may represent a new prognostic factor or a diagnostic biomarker in hemostasis research. PMID- 14533857 TI - N-methacryloly-(L)-histidinemethylester carrying a pseudospecific affinity sorbent for immunoglobulin-G isolation from human plasma in a column system. AB - N-methacryloly-(L)-histidinemethylester (MAH) as a pseudospecific ligand was synthesized by using methacryloyl chloride and histidine. Spherical beads with an average size of 63-75 microm were obtained by suspension polymerization of ethylene glycol dimethacrylate (EGDMA), 2-hydroxyethyl methacrylate (HEMA) and MAH conducted in an aqueous dispersion medium. The specific surface area of the beads was found to be 18.3 m2/g. Poly(EGDMA-HEMA-MAH) beads were used in the separation of immunoglobulin-G (HIgG) from aqueous solutions and/or human plasma in a packed-bed column system. HIgG adsorption capacity of the beads decreased with an increase in the flow-rate of plasma. The maximum HIgG adsorption on the poly(EGDMA-HEMA-MAH) sorbents was observed at pH 7.4. HIgG adsorption onto the poly(EGDMA-HEMA) sorbents was negligible. Higher adsorption values (up to 135 mg/g) were obtained when the poly(EGDMA-HEMA-MAH) sorbents were used from aqueous solutions. HIgG adsorption increased with decreasing temperature and the maximum adsorption achieved at 4 degrees C. MAH incorporation significantly affected HIgG adsorption capacity (135 mg/g). Higher amounts of HIgG were adsorbed from human plasma (up to 165 mg/g). Adsorption capacities of other blood proteins were obtained as 8.7 mg/g for fibrinogen and 14.6 mg/g for albumin. The total protein adsorption was determined as 191 mg/g. The pseudospecific affinity beads allowed one-step separation of HIgG from human plasma. HIgG molecules could be repeatedly adsorbed and desorbed with these sorbents without noticeable loss in their HIgG adsorption capacity. PMID- 14533858 TI - Synthesis, characterization and drug release from three-arm poly(epsilon caprolactone) maleic acid/poly(ethylene glycol) diacrylate hydrogels. AB - A biodegradable polymer network hydrogel with both hydrophobic and hydrophilic components was synthesized and characterized. The hydrophobic and hydrophilic components were a three-arm poly(epsilon-caprolactone) maleic acid (PGCL-Ma, as the hydrophobic constituent) and poly(ethylene glycol) diacrylate macromer (PEGDA, as a hydrophilic constituent), respectively. These two polymers were chemically photo-crosslinked to generate a three-dimensional network structure, which were characterized by FT-IR, DSC and SEM. The swelling property of the networks was studied in phosphate-buffered saline (PBS, pH 7.4). The results of this study showed that a wide-range swelling property was obtained by changing the composition ratio of PGCL-Ma to PEGDA. The in vitro release of bovine serum albumin (BSA) from these hydrogels as a function of the PEGDA to PGCL-Ma composition ratio and incubation time was examined and we found that the incorporation of PEGDA into PGCL-Ma increased the initial burst release of BSA. As the PEGDA component increased, the rate of formation of a loose three dimensional (3D) network structure increased; consequently, the sustained rate and extent of BSA release increased. We suggest that the release of BSA was controlled by both diffusion of BSA through swelling of the hydrophilic phase during an early stage and degradation of the hydrophobic phase during a late stage; and that the relative magnitude of diffusion versus degradation controlled release depended on composition ratio and immersion time. PMID- 14533859 TI - Hydrogel-microsphere-enhanced surface plasmon resonance for the detection of a K ras point mutation employing peptide nucleic acid. AB - Highly-sensitive detection of a K-ras point mutation in codon 12, frequently found in pancreatic cancer, based on DNA-carrying hydrogel microspheres as a response enhancer for surface plasmon resonance (SPR), is described. Acrylamide based microspheres with carboxyl groups were conjugated with DNA probes. Use of the DNA-carrying microsphere in the sandwich method, that is, binding of the microspheres with target DNAs at the sensor surface, enhanced the SPR response as a combined result of increased dielectric constant by the DNA-carrying microspheres. Microspheres lead to response enhancement, as shown by a 100-fold increase in sensitivity compared to that of non-amplified DNA target hybridization. In addition, the advantage of peptide nucleic acid (PNA) in the detection of a K-ras point mutation at the sensor surface by increasing temperature and flow rate is discussed. Results illustrate that the sandwich method through DNA-carrying microspheres for a SPR sensor is a promising approach for ultrasensitive DNA detection. PMID- 14533860 TI - A novel controlled drug-delivery system for growth hormone applied to healing skin wounds in diabetic rats. AB - Controlled release systems for drugs, hormones and growth factors can be particularly useful in tissue repair processes. These systems act as a biodegradable support containing the substance to be delivered, allowing their gradual release. In the past years, the local application of growth factors has acquired special relevance as a therapeutic option for use in subjects who show deficient tissue scarring, the hormone dose being the limiting factor for its success. In this study, the in vitro biocompatibility of a copolymer formed by vinylpyrrolidone and 2-hydroxyethyl methacrylate, used as an administration vehicle for hGH, was evaluated. The system was then tested in vivo in terms of its capacity for healing incisional wounds in healthy and diabetic rats. For the in vitro studies, polymer and hormone degradation rates were determined, and polymer biocompatibility was evaluated in fibroblast cultures. In the in vivo experiments, an incision was made in the back of the animals, and polymers discs with/without hGH, were introduced in the aperture. Morphological, immunohistochemical and morphometric evaluations were performed on wound tissue specimens 3-10 days after surgery. In vitro, the polymer was found to be biodegradable and showed no toxic effects on fibroblasts, the hormone being slowly released to the culture medium. In untreated diabetic rats, a delayed skin scarring and cell response were observed, compared to that noted in healthy animals. Skin closure, keratinisation and fibrosis occurred earlier in the presence of the polymer-hGH system. The use of this co-polymer as an administration vehicle for hGH improves the wound scarring process in the pathological setting of diabetes. PMID- 14533861 TI - Biodegradation of a dental composite by esterases: dependence on enzyme concentration and specificity. AB - Studies have shown that inflammatory (cholesterol esterase, CE) and salivary (pseudo-cholinesterase, PCE) enzymes can cause the breakdown of bisphenol-A diglycidyl dimethacrylate (bisGMA) and triethylene glycol dimethacrylate (TEGDMA) components from composite resins. Based on the above consideration, it was desired to show how CE- and PCE-catalyzed hydrolysis of resin components was dependent on the enzymes' concentration and to determine their distinct specificities (if any) towards resin components. Photopolymerized model composite resin samples (60% weight fraction silanated barium glass filler) based on bisGMA and TEGDMA monomers (55/45 weight ratio of the matrix, respectively) were incubated with PBS and either 0.01, 0.05, 0.1 or 1 unit/ml of CE or PCE for 16 days (pH 7.0, 37 degrees C). Incubation solutions were analyzed by high performance liquid chromatography (HPLC), UV spectroscopy and mass spectrometry. The composite samples were characterized by scanning electron microscopy (SEM). Degradation rates of bisGMA and TEGDMA monomers were assessed. The results showed that CE had a greater specificity towards cleaving bisGMA while PCE showed a greater specificity towards TEGDMA. A strong enzyme concentration dependence was observed which suggests that the level of degradation products generated for a material will depend on the esterase make-up of an individual's saliva in combination with the specific formulation of monomer components used. PMID- 14533862 TI - Self-aggregation behavior of alkylated chitosan and its effect on the release of a hydrophobic drug. AB - Chitosan (CS) was hydrophobically modified with butyl bromide and dodecyl bromide. The self-aggregation in acetic acid solution was characterized by fluorescence spectroscopy and dynamic light-scattering method. The results indicate that introducing butyl and dodecyl moieties leads to the formation of self-aggregates. Along with the enhancement in the hydrophobicity of chitosan the self-association occurs at a lower concentration, and the mean size of self aggregates increases. The loading capacity of butylated chitosan (4-CS) and dodecylated chitosan (12-CS) for vitamin B2 are markedly increased compared to that of chitosan, and the release of drug from alkylated chitosans is somewhat hindered due to its increased affinity for hydrophobic carriers. PMID- 14533863 TI - Thermal dehydration treatment and glutaraldehyde cross-linking to increase the biostability of collagen-chitosan porous scaffolds used as dermal equivalent. AB - A biodegradable scaffold for skin-tissue engineering was designed using collagen and chitosan, which are common materials for biomedical application. The scaffolds containing different amounts of chitosan were prepared by mixing the collagen and chitosan solutions followed by removal of the solvent using a freeze drying method. The cross-linking treatment of these scaffolds was performed using the dehydrothermal treatment (DHT) method or glutaraldehyde (GA) to increase their biostability. The effect of the chitosan concentration and the cross linking methods on the morphology of these scaffolds was studied by SEM. The water retention and the biodegradability in vitro of various collagen-chitosan scaffolds were investigated. Finally the biocompatibility of the collagen chitosan (10 wt% chitosan) scaffold treated with different cross-linking methods was evaluated using a in vivo animal test. A mild inflammatory reaction could be detected in the early stages, and GA treatment can decrease the inflammatory reaction in a long-term implantation. After implantation for four weeks, all kinds of scaffolds, especially the GA-treated scaffolds (Col-GA) were filled with a large number of fibroblasts and were vascularized to a certain extent. These results suggest that the GA-treated scaffold has an increased biostability and excellent biocompatibility. It can be a potential candidate for skin-tissue engineering. PMID- 14533864 TI - Safety assessment of Bt 176 maize in broiler nutrition: degradation of maize-DNA and its metabolic fate. AB - Insect resistant Bt 176 maize has been developed by genetic modification to resist European borer infection. In the present investigation, the experiment was conducted to determine the effect of feeding a new hybrid of Bt 176 maize (NX 6262- Bt 176) on general health condition and performance of broiler chickens. Maize grains and diets were subjected to proximate analysis. Amino and fatty acids investigation were applied for both maize grains before used. To evaluate the degradation of NX 6262- Bt 176 maize DNA and its metabolic fate in broiler blood, muscles and organs. One-day-old male broilers were fed ad libitum on either an experimental diet containing NX 6262- Bt 176 or a control diet containing the non-modified maize grains for 35 days. Feed consumption and body weight were recorded weekly during the experimental period. All chickens were subjected to nutritional evaluation period at day 20 of age for 5 successive days, to calculate the percentage of apparent digestible nutrients in both diets. At day 35 samples were collected at several intervals after feed withdrawal. Prior to slaughter blood samples were collected from all birds by heart puncture to prevent DNA cross contamination. Samples from pectoral and thigh muscles, liver, spleen, kidney, heart muscle, bursa and thymus glands were collected. Digesta from different sections of the gastrointestinal tract (GIT) were collected as well. Packed cell volume (PCV) and some serum parameters were investigated. There were no significant differences between control and experimental group concerning chemical composition of feeds, apparent digestible nutrients, and all performance parameters measured (P > 0.05). Furthermore, there were no differences in the PCV and the analysed serum parameters between the control and experimental group. The results of maize DNA digestibility showed that the new variety takes the normal physiological passage along broiler GIT similar to the conventional line. In addition, Bt 176 maize DNA appears to be partially degraded in different parts of GIT comparable to the DNA of the control maize line. Results of the metabolic fate of maize DNA in broiler blood, muscles and organs indicated that only short DNA fragments (199 bp) derived from the plant chloroplast gene could be detected in the blood, skeletal muscles, liver, spleen and kidney, which disappeared after prolongation the fasting time. In heart muscle, bursa of Fabricius and thymus, no plant chloroplast DNA was found. Bt gene specific constructs from Bt 176 maize were not detected in any investigated blood or tissue samples. PMID- 14533865 TI - Nutrient digestibility and prediction of metabolizable energy in total mixed rations for ruminants. AB - The aim of the present study was to determine equations that predict ME in total mixed rations (TMR) based on routine methods. The ME content of 30 TMR for dairy cows was determined based on digestible crude nutrients obtained with wether sheep. Concentrations in the TMR (in g/kg DM) varied between 118 and 234 for crude protein, 26 and 48 for crude lipid, 131 and 250 for crude fibre, 281 and 488 for NDF, and 173 and 304 for ADF. Gas production ranged from 40.7 to 54.1 ml/200 mg DM, and enzymatically degraded organic matter from 652 to 800 g/kg DM. Digestibility [%] ranged from 68.6 to 84.0 for organic matter, from 55.6 to 84.3 for crude lipid, from 55.0 to 77.8 for crude fibre, from 57.6 to 77.0 for NDF and from 53.1 to 79.6 for ADF. ME ranged from 9.6 to 11.9 MJ/kg DM, and NEL from 5.7 to 7.4 MJ/kg DM. ME content was highly correlated with the concentration of both crude fibre and enzymatically degradable organic matter as well as with organic matter digestibility. A multiple regression equation based on crude fibre and crude lipid predicted ME with a reasonable goodness of fit (r2 = 0.81; s(y.x) = 2.4%). The inclusion of other nutrients, of neutral and acid detergent fibre, neither of gas production did improve the goodness of fit. The best prediction was achieved with inclusion of enzymatically degraded organic matter (r2 = 0.90; s(y.x) = 1.7%). PMID- 14533867 TI - Effects of protozoa on methane production in rumen and hindgut of calves around time of weaning. AB - Effects of the presence or absence of ciliate protozoa on methanogenesis in the rumen and hindgut were investigated in young calves during a 7-week period. Ten Holstein calves, aged 7 days, were divided in two groups (n = 5) and fed an increasing amount of a commercial milk replacer and small amounts of a calves starter. One group was inoculated with ciliate fauna on two occasions, week 5 and 6, while the second remained ciliate-free. The absence of protozoa in the rumen decreased rumen empty weight (-23%, P < 0.01), and rumen pool size of N (-36%, P < 0.01) and crude fat (-37%, P < 0.05). Rumen bacteria of non-faunated calves contained a higher proportion of total amino acid-N per 16 g N (+3%, P < 0.01) and D-alanine-N per 16 g N (+13%, P < 0.05) compared to faunated calves. Further results contain a reference for a higher bacterial mass in the ciliate-free rumen with an increased number of bacteria adherent to rumen mucosa. The CH4 production in the rumen increased exponentially with the increase in protozoa population size (R2 = 0.68). In presence of 46 x 10(4) protozoa per ml rumen fluid, the in vitro CH4 production of rumen fluid per mol total VFA was about 34% higher in faunated than in non-faunated calves (P < 0.001). Hydrogen (2H) recovery of rumen fermentation was positively correlated (R2 = 0.55) to the CH4 production rate. Methanogens were attached on rumen mucosa. Methanogenesis, induced by rumen mucosa attached bacteria, was stimulated by ruminal protozoa. In the absence of protozoa in the rumen, the acetate-propionate ratio and butyrate proportion of VFA were reduced. In vivo, in the absence of protozoa not only the whole animal CH4 production (-30%, P < 0.05) but also the digestibility of carbohydrates (-4%, P < 0.05) was reduced. Thereby no difference was observed in the intake of ME per kg DM between the groups. In conclusion, the methanogenesis in the rumen, but not in hindgut, is associated with the development of the ruminal protozoa population. The level of methanogenesis (mol/mol VFA) in the hindgut amounts to 20% of the ruminal methanogenesis. PMID- 14533866 TI - Anti-nutritional effects of a moderate dose of soybean agglutinin in the rat. AB - This study was conducted to investigate the effects of a moderate dose of purified soybean agglutinin on performance and nitrogen digestibility in rats as well as to determine its effects on the protein, DNA and RNA content of the small intestine and pancreas. Twenty-four Sprague-Dawley rats were randomly allotted into one of four groups for a 10-day nitrogen balance experiment. The four groups of rats were fed 7 g of a casein-cornstarch based diet or a similar diet supplemented with 0.1, 0.2 or 0.4 mg/g purified soybean agglutinin. All experimental diets were adjusted to an identical nutrient level. Dose of soybean agglutinin had no significant effect on rat performance. Incorporation of soybean agglutinin in the diet reduced apparent protein digestibility and the utilization of dietary protein by increasing nitrogen loss from the faeces and urine. Fresh pancreatic weight increased in rats fed soybean agglutinin at a level of 0.4 mg/g in the diet compared to the control, but the dry pancreatic weight and the protein content of the pancreas did not differ among the four groups. However the DNA and RNA content of the pancreas had a tendency to increase with a higher level of soybean agglutinin. The weight of the jejunum and its protein, DNA and RNA content were not significantly affected by soybean agglutinin, but the dry weight and the RNA of the jejunum tended to increase with higher levels of soybean agglutinin in the diet. In conclusion, purified soybean agglutinin, at moderate levels in the rats diet, had negative effects on digestive function, such as nitrogen digestibility, nitrogen retention and nitrogen balance. As the level of soybean agglutinin increased, the effects became more pronounced. Meanwhile, hypertrophy of the pancreas was observed with higher doses of soybean agglutinin incorporation in the diets. PMID- 14533868 TI - Apparent ileal digestibility of amino acids and the digestible and metabolizable energy content of high-oil corn varieties and its effects on growth performance of pigs. AB - Two experiments were conducted to compare the nutritional value of normal and high-oil corn for pigs. The normal corn and the two varieties (high-oil corns A and B) of high-oil corn contained 4.41, 7.35 and 8.86% ether extract, on DM basis, respectively. In experiment 1, six non-littermate crossbred barrows (37.8 +/- 1.3 kg BW) were fitted with ileal T-cannulas and used in a double replicated Latin Square digestion trial. Three diets were formulated containing 96.6% of one of the three varieties of corn as the only protein source. Chromic oxide (0.4%) was added as a digestibility marker. Additional vitamins and minerals were added to meet requirements. The digestible energy concentrations for normal corn and high-oil corn A and B were 16.53, 16.99 and 17.07 MJ/kg while the metabolizable energy values were 15.82, 16.32 and 16.36 MJ/kg, on DM basis, respectively. The ileal amino acid digestibility of high-oil corn was generally higher than that of normal corn with significant differences being observed for the essential amino acids isoleucine and phenylalanine. In experiment 2, 96 pigs (8.01 +/- 0.14 kg BW) were used to evaluate four diets in a 2 x 2 factorial design conducted over a 35-day period. Corn variety (high-oil vs. normal corn) and nutrient density (high content of protein and ME vs. low content of protein and ME) were set as the two main effects. During the first 14 days, pigs fed high-oil corn diets consumed more feed and tended to get higher daily gain than pigs fed normal corn. Over the entire 35-day experiment, increasing dietary nutrient density increased daily gain and tended to increase feed conversion, while variety of corn had no significant effects on performance. Overall, the present results indicate that the energy concentration and ileal amino acid digestibility of high-oil corn varieties were equal or superior to those in normal corn and therefore they should be able to be effectively utilized in diets fed to swine. PMID- 14533869 TI - Research note: changes in the concentration of beta-carotene, alpha-tocopherol and retinol in the bovine corpus luteum during the ovarian cycle. AB - Causes and possible consequences of the accumulation of beta-carotene during the development of the corpus luteum in cattle are still unknown. Therefore, in a descriptive study a total of 43 corpora lutea from ovaries of non-pregnant cows representing four stages of the ovarian cycle the concentration of beta-carotene, alpha-tocopherol and retinol was determined. Beta-carotene and alpha-tocopherol continuously increased from stage I to stage IV (beta-carotene: 14 +/- 8 vs. 175 +/- 117 microg/g, P < 0.05; alpha-tocopherol: 15 +/- 7 vs. 132 +/- 66, P < 0.001). In contrast, retinol decreased from highest values at stage I (250 +/- 160 ng/g) to lowest (90 +/- 80 ng/g tissue) at stage III (P < 0.05). The results support the concept, that both beta-carotene and alpha-tocopherol accumulation in the corpus luteum secondary to the uptake of lipoprotein-bound cholesterol during steroid synthesis. Lowest retinol levels were observed at highest metabolic activity of the corpus luteum indicating a possible consumption of retinol during stereogenesis. In conclusion assuming an importance of a local conversion of beta carotene into retinol in the corpus luteum dietary supplementation should be sufficient enough to support such an accumulation of beta-carotene in the corpus luteum as local precursor of retinol. PMID- 14533870 TI - Lack of efficacy of the combination of pamidronate and vitamin D on regression of prostate cancer in the Dunning rat model. PMID- 14533871 TI - Molecular regulation of cholesterol biosynthesis: implications in carcinogenesis. AB - Cholesterol synthesis was demonstrated to be mandatory for cellular growth and serves to supply one of the necessary building blocks for new membranes demanded by dividing cells during growth. The mevalonate pathway, which is regulated through a finely tuned mechanism, is responsible mainly for cholesterol enrichment to cells. Among the various steps, the production of mevalonate from 3 hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) is the most critically regulated step catalyzed by HMG-CoA reductase. The ability of sterols to regulate both the transcriptional rates of the reductase gene and the degradative machinery for the reductase protein provides a multilevel system for controlling the expression of this enzyme. Much convincing evidence indicates that cells manifest a higher flux through the mevalonate pathway when proliferating than when they are in the cell cycle arrest condition; furthermore, tumors undergo deregulated cholesterogenesis mainly at the critical rate-controlling juncture (i.e., the reaction catalyzed by HMG-CoA reductase). The mevalonate component of the cholesterol biosynthesis plays a key role in controlling cell proliferation by generating prenyl intermediates, particularly farnesyl and geranyl-geranyl moieties. These isoprenoids covalently modify and thus modulate the biological activity of signal transducing proteins, such as that of the Ras superfamily. The prenylated Ras mediated signal transduction pathway provides much of the molecular information needed to trigger cell proliferation. Therefore, depletion of mevalonate can block both the processing and the transforming activities of Ras, indicating that drugs such as lovastatin and compactin, which had previously been exploited for lowering cholesterol levels, may be useful chemotherapeutic agents for treating tumors harboring oncogenic Ras mutation. In addition, Ras prenylation, which provides much of the molecular information needed to trigger cell proliferation, represents an inviting target for the design of chemotherapeutic drugs that would interrupt such signaling events and arrest tumor cell proliferation. PMID- 14533872 TI - Membrane-associated MMP-2 in human cervical cancer. AB - Matrixmetalloproteinases (MMPs) are a family of secreted or membrane-associated proteins capable of digesting extracellular matrix components. The importance of MMPs in tumor development and invasion is well known. Recent studies have strongly indicated the presence of a functional complex consisting of alpha(v)beta3 integrin, membrane type-1 metalloproteinase-2 (MT1-MMP), and tissue inhibitor of metalloproteinase-2 (TIMP-2) on tumor cell surface, which helps the activation of MMP-2. In this article, we report on the association of active MMP 2 with the membrane fraction of human cervical cancer cells. The association of MMP-2 with alpha(v)beta3 integrin and the expression of membrane type MT1-MMP strongly indicate the role of alpha(v)beta3/MT1-MMP/TIMP-2 complex in the activation of MMP-2 in cervical cancer tissue membrane fraction. The membrane associated activated MMP-2 may have a role in the migration of tumor cells during invasion. PMID- 14533873 TI - Genotoxicity of textile dyes evaluated with Ames test and rec-assay. AB - We evaluated for mutagenicity 14 commercial textile dyes used extensively in the northern part of India using both the Ames Salmonella typhimurium microsome reversion test as well as the recombination-repair (rec)-assay. The Ames test revealed that 57.14% of dyes were mutagenic and acting directly. The rec-assay detected 50% of dyes to be mutagenic; of these, 71.43% were direct acting, whereas 28.57% required Aroclor-induced exogenous metabolic activation. Used together, the two tests detected 78.57% of the dyes to be mutagenic, and a 50% correlation was found between these two tests. Groupwise, three out of four azo dyes and all five anthraquinone dyes were found to be mutagenic by the Ames assay; the rec-assay detected methine/polymethine (1 out of 3), an oxazine, and a triphenylmethane dye to be mutagenic, besides the azo (1 out of 4) and the anthraquinone (3 out of 5) dyes. The structure-activity analysis attributed the mutagenicity of dyes to the structural alerts such as phenylenediamine, amino and nitro-groups, methylation, CH=CH, and chloro groups; whereas deamination, bulkier groups, and sulfonation may be responsible for diminishing mutagenicity. PMID- 14533874 TI - Expression of cAMP-responsive element modulator (CREM) in rat testes following chronic cocaine administration. AB - OBJECTIVE: c-AMP-responsive element modulator (CREM), one of the nuclear factors involved in the regulation of gene expression by cAMP, has an important role in spermatogenesis. Our recent study has shown that chronic administration of cocaine to male rats results in disruption of spermatogenesis, including reduction of germ cells. As a further step toward understanding this process, we have studied the role of CREM in cocaine-induced testicular damage. MATERIALS AND METHODS: Sprague-Dawley rats were administered cocaine hydrochloride subcutaneously daily for 90 days. Control animals received equal volumes of normal saline daily for 90 days. Testes were removed after 15, 30, 90 days of cocaine administration. Total RNA was extracted from the testes and subjected to RT-PCR. Testicular tissue was also homogenized in a lysis buffer, and Western blotting was performed using anti-CREM antibody. RESULTS: RT-PCR analysis detected a single fragment of approximately 520 base pairs (bp) in control testes at all time points. The cocaine-treated testes showed reduced expression of CREM fragment. Western blot analysis using CREM antibodies confirmed the RNA data. There were reduced CREM proteins in the cocaine-treated testes compared with controls. CONCLUSIONS: The CREM gene is essential for spermatogenesis. Our results indicate that the reduction in testicular CREM expression may be one of the mechanisms responsible for disruption or impairment of spermatogenesis in the testes following chronic cocaine administration. PMID- 14533875 TI - The role of pteridines in physiology and pathology. AB - Disorders or persistent noxious stimulation of the neuroimmunological circuitry can lead to neoplastic, neurological, immunological, psychiatric, or multiorgan pathology. The cause/effect relationship has encouraged a search for neuroimmunological markers possessing functional or pathological correlates. Among the molecules studied, those associated with the biopterin biosynthetic pathway (including neopterin) have proven to be useful and informative. This review addresses the relevance of the latter mediators to our understanding of the pathophysiology of neoplastic, neurological, psychiatric, and immunological disease processes. PMID- 14533876 TI - Dietary cured meat and the risk of adult glioma: a meta-analysis of nine observational studies. AB - OBJECTIVE: N-nitroso compounds (NOCs) are recognized neural carcinogens in animal models and are suspected human carcinogens. A meta-analysis was performed examining the possible association of dietary N-nitroso intake from cured meats and the risk of gliomas in adults. METHODS: A prospective protocol was developed outlining the intent, methods, and statistical analysis of the meta-analysis. Data from nine epidemiological studies were pooled using a general variance-based meta-analytic method employing confidence intervals as described by Greenland. The outcome of interest was a summary relative risk (SRR) reflecting the risk of brain tumor (glioma) development associated with high dietary intake of cured meats (as defined by individual study investigators). Sensitivity analyses were performed when necessary to explain any observed statistical heterogeneity. RESULTS: Nine observational studies met protocol-specified inclusion criteria. Analysis for heterogeneity demonstrated a lack of statistical heterogeneity (p = 0.58). Pooling the data on dietary cured meat intake of all types yielded an RR of 1.48 (1.20-1.83), suggesting a 48% increased risk of glioma development among adults ingesting high levels of cured meat. Analyzing brain tumor risk by meat type yielded an RR of 0.90 (0.63-1.25) for hotdogs (a nonstatistically significant result), 1.31 (1.00-1.71) for bacon, and 1.64 (1.27-2.14) for ham. Sensitivity analyses showed that the failure of most studies to adjust for total energy intake might lead to a spurious positive association between cured meat intake and brain tumor risk. Insufficient data were available for analyzing dose response relationships, although a few individual studies showed evidence of increasing risk with increasing cured meat intake. CONCLUSION: The available data do not provide clear support for the suspected causal association between ingestion of NOCs from cured meat in adults and subsequent brain tumor risk. Uncontrolled confounding may account for the previously noted positive association seen in some epidemiological studies. PMID- 14533877 TI - Perioperative immunosuppression in cancer patients. AB - Cancer patients commonly undergo surgical procedures. The perioperative period is characterized by immunosuppression and may predispose already immunosupressed cancer patients to tumor spread. Cancer patients typically show depression of both cellular and humoral immune functions. Possible mediating factors for immunosuppression during the perioperative period include anesthetic agents, opioids, surgery, blood transfusions, temperature changes, pain, and psychological stress. A surgically mediated decrease in natural killer (NK) cell activity has been implicated as the major contributing factor associated with an increase in metastasis. The decreased NK cell activity during the perioperative period is associated with increased risk of mortality and cancer. Commonly used anesthetic agents and opioids are known to inhibit NK cell activity. Despite the in vivo evidence of anesthetic- and analgesic-agent-mediated immunosupression, surgery by itself results in a three- to four-fold increase in retention of metastasis when compared to the groups in which anesthesia and analgesia were combined. The negative consequences associated with perioperative immunosuppression may be decreased by several strategies, including aggressive pain control, selection of specific anesthetic and analgesic agents, avoidance of unnecessary transfusions, and delay of elective surgeries until the patient's nutritional and immune status is optimized. Recognizing and neutralizing its mediating factors, perioperative immunosuppression in cancer patients may be reduced. PMID- 14533878 TI - A convenient method for the synthesis of 2-(beta-D-glycopyranosylthio) pyridines. AB - Treatment of piperidinium salts of dihydropyridinethiolates 3 with glycosyl bromides 4 in dry acetone provides a convenient and high yielding synthesis of 1,4-dihydro-3-cyanopyridine thioglycosides 5. The structures of 5 were confirmed by oxidation as well as by 1H NMR and 13C NMR spectral analysis. PMID- 14533879 TI - Open-chain carbocyclic analogs of adenosine with dihalovinyl unit as potential inhibitors of S-adenosyl-L-homocysteine hydrolase. AB - Vinylogously extended deoxyeritadenine derivatives were synthesized as acyclic/ carbocyclic analogues of the 6'-halo(homovinyl)adenosines, which are known to be potent inhibitors of S-adenosyl-L-homocysteine hydrolase. Swern oxidation of 9-[3 (t-butyldimethylsilyloxy)-4-hydroxybutyl]adenine (4) followed by Wittig olefination and desilylation gave access to ethyl 6-(adenin-9-yl)-4-hydroxy-2(E) hexenoate (7) and 5-(adenin-9-yl)-1,1-dibromo-1-penten-3-ol (9). No inhibition of AdoHcy Hydrolase was observed with 7 and 9. PMID- 14533880 TI - Characterization and elucidation of coordination requirements of adenine nucleotides complexes with Fe(II) ions. AB - In spite of the significant role of iron ions-nucleotide complexes in living cells, these complexes have been studied only to a limited extent. Therefore, we fully characterized the ATP:Fe(II) complex including stoichiometry, geometry, stability constants, and dependence of Fe(II)-coordination on pH. A 1:1 stoichiometry was established for the ATP:Fe(II) complex based on volumetric titrations, UV and SEM/EDX measurements. The coordination sites of ferrous ions in the complex with ATP, established by 1H-, 31P-, and 15N-NMR, involve the adenine N7 as well as P(alpha), P(beta), and P(gamma). Coordination sites remain the same within the pH range of 3.1-8.3. By applying fluorescence monitored Fe(II)-titration, we established a logK value of 5.13 for the Fe(ATP)2- complex, and 2.31 for the Fe(HATP)-complex. Ferrous complexes of ADP3- and AMP2- were less stable (log K 4.43 and 1.68, respectively). The proposed major structure for the Fe(ATP)2- complex is the 'open' structure. In the minor 'closed' structure N7 nitrogen is probably coordinated with Fe(II) through a bridging water molecule. The electronic and stereochemical requirements for Fe(II)-coordination with ATP4- were probed using a series of modified-phosphate or modified-adenine ATP analogues. We concluded that: Fe(II) coordinates solely with the phosphate-oxygen atom, and not with sulfur, amine, or borane in the cases of phosphate-modified analogues of ATP; a high electron density on N7 and an anti conformation of the adenine-nucleotide are required for enhanced stability of ATP analogues:Fe(II) complexes as compared to ATP complexes (up to more than 100-fold); there are no stereochemical preferences for Fe(II)-coordination with either Rp or Sp isomers of ATP-alpha-S or ATP-alpha-BH3 analogues. PMID- 14533881 TI - Novel synthesis of 4'C-aryl-branched acyclic nucleoside using [3,3]-sigmatropic rearrangement. AB - A very efficient synthetic route for preparing a novel 4'-C-aryl branched-1',2' seco-2',3'-dideoxy-2',3'-didehydro-nucleoside is described. Mesylate 7 was successfully synthesized via a Horner-Wadsworth-Emmons reaction and a [3,3] sigmatropic rearrangement, with which an adenine base was coupled by nucleophilic substitution conditions (K2CO3, 18-Crown-6, DMF) to give the target nucleoside 9. PMID- 14533882 TI - 3'-Azido-3'-deoxy-5'-O-isonicotinoylthymidine: a novel antiretroviral analog of zidovudine. II. Stability in aqueous media and experimental and theoretical ionization constants. AB - Degradation of 3'-azido-3'-deoxy-5'-O-isonicotinoylthymidine (AZT-Iso), an antiretroviral derivative of zidovudine, was investigated in buffer pH 7.4, mu = 300 mOsm at 37, 50 and 60 degrees C, and in water (pH 6.6, 37 degrees C), giving zidovudine (AZT) and isonicotinic acid (INA) as products. The rate constants were determined by reversed-phase HPLC showing pseudo-first-order kinetics related to the residual amount of AZT-Iso. In this way, the studied compound was demonstrated to be 153 times more stable in water than in buffer solution at 37 degrees C. The analytical method was conveniently validated demonstrating to be a rapid and accurate stability-indicating technique. In addition, experimental and theoretical values of pKa were determined. PMID- 14533883 TI - Synthesis of some new 2-alpha-L-arabinopyranosyl-1,2,4-triazines as potential antitumor chemotherapeutics. AB - Synthetic routes towards different 2-alpha-L-arabinopyranosyl-3-thioxo-2,3 dihydro-1,2,4-triazin-5(4H)-/ones or thiones were investigated. Primary human anticancer screening of two selected compounds resulted in an active compound against SF-268 (CNS) cell line. PMID- 14533884 TI - Synthesis of some N-galactosides of 3-aryl-5-benzyl (or substituted benzyl)-1,2,4 triazin-6(1H)-/ones or thiones of expected biological activity. AB - The 1-(2,3,4,6-tetra-O-acetyl-beta-D-galactopyranosyl)-3-aryl-5-benzyl (or substituted benzyl)-1,2,4-triazin-6(1H)-/ones or thiones were prepared via galactosidation of 3-aryl-5-benzyl (or substituted benzyl)-1,2,4-triazin-6(1H) /ones or thiones with 2,3,4,6-tetra-O-acetyl-alpha-D-galactopyranosyl bromide. The structure of the new galactosyl derivatives was based on both spectroscopic and chemical evidences. PMID- 14533885 TI - Permanganate oxidation reactions of DNA: perspective in biological studies. AB - KMnO4 has been well known as a powerful chemical probe for numerous applications in biological fields, particularly for those used in conformational studies of DNA. The KMnO4 assay provides essential information for understanding biochemical processes and detecting aberrant DNA, which is associated with many genetic diseases. Elegant examples are sequencing techniques, foot-printing assays for transcriptional studies, an interference method for hormone receptor binding assays as well as DNA conformational studies of Z-DNA, Z-Z junctions, hairpins, curvatures, short nucleotide base repeats, binding of intercalators and groove binders, etc. Recently, KMnO4 has been successfully applied to detect single base changes and mutations in DNA (chemical cleavage of mismatch method, CCM) as well as other types of base damage (8-oxoguanine and thymine dimers). This paper aims to review the usefulness and limitations of the permanganate oxidation reaction used in various biological studies of DNA. PMID- 14533886 TI - Maternal-fetal surgery. PMID- 14533887 TI - In their footsteps: a brief history of maternal-fetal surgery. AB - These four individuals are recognized because at a critical point in the evolution of maternal-fetal surgery they contributed in ways that uniquely advanced progress in the field, enabling other researchers to make even greater discoveries. They were not the only significant contributors; there were many, too numerous to recount. Although their contributions were crucial, other researchers accomplished more. Among their peers, however, these four pioneers stand out because at a critical nexus their vision and perseverance carried researchers forward to a new level of excellence. We all follow in their footsteps. PMID- 14533888 TI - Ethics: "life before birth" and moral complexity in maternal-fetal surgery for spina bifida. AB - This article considers the ethical significance of a moral belief common among pregnant women (and their partners) who seek open uterine repair for fetal spina, namely that their fetuses are already "babies." The need to recognize and interact sensitively with a pregnant woman's vulnerability to her own beliefs and concerns regarding potential disabilities, the fetal intervention, and its potential outcome is emphasized. Such recognition and explicit discussions are ethically important for informed consent and to safeguard against the judgments, enthusiasms, and biases of surgeons and other team members. PMID- 14533889 TI - Congenital diaphragmatic hernia: advances in prenatal therapy. AB - Despite intensive clinical and experimental efforts, mortality from CDH remains high. More than two decades of research in multiple centers has led to a better understanding of the pathophysiology, prognosis, and treatment options for fetuses that have CDH. It now appears that fetuses that have prenatally diagnosed CDH can be stratified into high- and low-risk groups based upon sonographic parameters. Fetuses that do not have liver herniation into the chest that have a favorable LHR have an excellent chance of survival with postnatal therapy. Prenatal diagnosis allows the time and place of delivery to be planned in advance so these infants can be treated in a tertiary care nursery that has maximal medical and surgical therapy. Fetuses that have liver herniation into the chest and an unfavorable LHR have a grim prognosis. These fetuses might benefit from in utero intervention. There is no role for open fetal repair of the diaphragmatic detect; however, fetoscopic temporary tracheal occlusion might improve lung growth and development and might decrease morbidity and mortality in these infants. The FETENDO strategy appears to work, and for the first time it offers hope to the fetus that has high-risk CDH, but its efficacy must be proven in a proper randomized, controlled trial. PMID- 14533890 TI - Management of fetal lung lesions. AB - We have learned from prenatal diagnosis that there is a wide spectrum of clinical severity for fetuses that have a lung mass. Accurate prognostic information is necessary for providing appropriate management and parental counseling. If an associated life-threatening anomaly is present or if the mother is ill with the mirror syndrome, then the family might choose to terminate the pregnancy. If the fetus is not hydropic and an isolated fetal lung lesion is present, the mother is followed by serial ultrasound and arrangements are made for the best possible care after birth. Some CCAMs and many BPSs will shrink in size, so it is important to try to differentiate these lesions using prenatal diagnostic criteria, although this technique is not always possible. All fetuses that had fetal thoracic masses without hydrops in our series survived in the setting of maternal transport, planned delivery, and postnatal evaluation at a facility with ECMO capability. Many of the babies that had large lesions at our center required ventilatory support, and six babies needed treatment with ECMO. Our impression is that these nonhydropic fetuses that had lung masses had less lung hypoplasia and a much better prognosis than those that had diaphragmatic hernia despite a similar degree of mediastinal shift as judged by prenatal sonography. In asymptomatic neonates that have a cystic lung lesion, we believe that elective resection is warranted because of the risks of infection and occult malignant transformation. Malignancies consist mainly of pleuropulmonary blastoma in infants and young children and bronchioloalveolar carcinoma in older children and adults. After confirmation of CCAM location by postnatal chest CT scan with intravenous contrast, we recommend elective resection at 1 month of age or older. This age has been chosen because anesthetic risk in babies decreases after 4 weeks of age. An experienced pediatric surgeon can safely perform a lobectomy in infants with minimal morbidity. Early resection also maximizes compensatory lung growth. In contrast, we have usually followed patients with a tiny, asymptomatic, noncystic BPS if we are confident of the diagnosis based on postnatal imaging studies. We do not favor the approach of catheterization and embolization for the treatment of larger BPS lesions. If the fetus is hydropic at presentation or if hydrops develops during serial follow-up, management depends upon the gestational age. For hydropic fetuses of greater than 32 weeks' gestation, early delivery should be considered so that the lesion can be resected ex utero, but the neonatal outcome is dismal. We recently managed two such cases using an ex utero intrapartum therapy (EXIT) strategy with resection of the mass during the EXIT procedure. Both fetuses survived and one required the use of ECMO. For hydropic fetuses of less than 32 weeks' gestation, there is now a new therapeutic option, treating the lesion before birth. PMID- 14533891 TI - Fetal surgery for sacrococcygeal teratoma. AB - A great deal has been learned regarding the natural history and pathophysiology of fetal SCT. The logic behind fetal intervention for SCT and hydrops appears to be correct, and open and minimal access techniques of fetal intervention have been shown to be feasible. The development of fetal intervention for SCT has mirrored those developed for other diseases such as congenital diaphragmatic hernia. In a recent presentation, Harrison, the original pioneer in fetal surgery, outlined trends in fetal intervention. The first trend is that of moving from open, invasive techniques to minimally invasive techniques. In the case of SCT surgeons are moving from open resection to RFA and possibly to fetoscopic resection. The second trend outlined by Harrison is a movement away from total in utero repair of a defect that recapitulates postnatal treatment and toward manipulation of fetal pathophysiology to reverse life-threatening events. In SCT surgeons employ RFA to ablate causative blood vessels to reverse fetal hydrops with the knowledge that these fetuses will require postnatal resection of the tumor. In contrast to resection, RFA requires less time and significantly less maternal morbidity than open resection. Further study is required to determine the role of minimal access techniques in SCT. Future directions for treatment of fetal SCT with hydrops might include fetoscopic resection or high-intensity ultrasound ablation. PMID- 14533892 TI - Distal urinary obstruction. AB - Despite the sound experimental basis and initial promise of early animal models, the results of antenatal intervention have been disappointing, with high rates of misdiagnosis of urethral valves, complications from vesicoamniotic shunting, perinatal mortality, and long-term renal impairment and bladder dysfunction in survivors. The recent development of a cystoscopic approach might obviate some of these problems, but to date the procedure been limited by technical difficulty in negotiating the urethrovesical angle. Overcoming these difficulties through equipment modifications might allow definitive testing of whether or not alleviating distal urinary obstruction in utero is beneficial. PMID- 14533893 TI - Intrauterine myelomeningocele repair. AB - Preliminary studies have suggested that IUMR reduces the incidence of shunt dependent hydrocephalus and the severity of the Chiari malformation. An RCT is in progress to confirm these results. Future efforts will revolve around refinement of the procedure with the goal of reducing risk. Robot-assisted surgery holds the promise of achieving this goal. If it is successful. IUMR might well become the standard therapy for myelomeningocele, resulting in a significant reduction in the devastating morbidity associated with this disease. PMID- 14533894 TI - Fetal hydrocephalus. AB - Fetal hydrocephalus is a dynamic process. Its natural progression is not completely understood, but it is almost always associated with other intracranial and extracranial anomalies. Fetal hydrocephalus might begin as mild ventriculomegaly and progress to a more serious state. Mild ventricular enlargement without an increase in head size might be a normal variant. Careful prenatal evaluation is always indicated when the diagnosis of ventricular enlargement has been made, and it should include a detailed anatomic survey and genetic amniocentesis. Therapeutic options need to be discussed in detail with the patient. If multiple associated anomalies--especially other CNS anomalies- are present, the prognosis for a viable outcome is dismal. If isolated hydrocephalus is seen, gender determination is appropriate in the consideration of X-linked hydrocephalus secondary to aqueductal stenosis. Communicating hydrocephalus can also be mistaken for obstructive ventriculomegaly, but it can usually be excluded by demonstration of dilation of the subarachnoid cistern. There is a small subset of fetuses that have isolated hydrocephalus that might benefit from prenatal surgical intervention. Feasibility studies are in progress to assess the therapeutic benefit of this type of therapy. Gestational age requirements and strict criteria are being evaluated. As with the multiple therapies discussed in this issue, many interventions are becoming more and more feasible, and antenatal surgery is likely to be limited only by the ability to think imaginatively and to act creatively. PMID- 14533895 TI - Balloon valvuloplasty for congenital heart disease in the fetus. AB - Fetal cardiac interventions are relatively new and promising therapeutic options for modifying CHD in utero. Techniques for safe access to the fetus must be improved and patient selection criteria must be developed. Most important is early detection and referral of all patients who have CHD, enabling improved outcomes for infants. Finally, it should be stressed that performing successful fetal cardiac interventions requires multidisciplinary collaboration between perinatologists, cardiologists, fetal surgeons, and anesthesiologists. Each discipline has specific skills for providing safe access to the fetus, performing the procedure, and providing perioperative care. PMID- 14533896 TI - Prenatal management of gastroschisis: the place of the amnioexchange procedure. AB - Gastroschisis is a malformation of the anterior abdominal wall that consists of a right paraumbilical defect with bowel loops bathed in the amniotic fluid. The survival rate is now greater than 90% and the prognosis relies mainly on morbidity attributable to bowel dysfunction. Recent research has examined gastrointestinal waste present in amniotic fluid that induces bowel toxicity and an inflammatory process. The amnioexchange procedure (changing the amniotic fluid regularly) involves a new therapeutic approach: reducing bowel injuries in the fetuses. This article shows that there is an inflammatory reaction in human gastroschisis and in the authors' model, and that the clinical and biological data plead for the practice of amnioexchange in human beings. A randomized, controlled study is now needed. PMID- 14533897 TI - Treatment of previable premature ruptured membranes. AB - We have gained significant experience in the clinical understanding and treatment of PPROM in the last 5 years. From our work we can conclude that iatrogenic PPROM is not only a different entity etiologically, it is also different than spontaneous PPROM in its clinical behavior and response to therapeutic measures. Our current success rate with the amniopatch allows us to suggest that iatrogenic PPROM should no longer be viewed as a devastating complication of pregnancy. Instead, as other investigators corroborate our successful experience with the amniopatch, this procedure might become standard treatment for this unusual but frustrating complication of prenatal intervention. Regarding spontaneous PPROM, our work suggests that perhaps, in a select group of patients (in whom infection has not set in and in whom a smaller and better defined membrane defect might exist), there is the potential for developing a surgical technique that might graft the defect successfully and allow the integrity of the amniotic membrane to be restored. The actual surgical technique, the materials to be used, and the approach (transabdominal versus transcervical) are areas of research for the future. Meanwhile, continued efforts are warranted to decipher whether or not this obstetrical complication can one day be conquered. PMID- 14533898 TI - Twin-twin transfusion syndrome. AB - The understanding and management of twin-twin transfusion syndrome has evolved significantly over the past few years. Improved and standardized sonographic diagnostic criteria, understanding of the heterogeneic nature of the syndrome, development of an anatomical and reproducible surgical technique for the identification of vascular anastomoses, and technological advances and developments now allow clinicians to view the disease as a more readily understandable and treatable condition. Many tasks remain, including education of peers, better screening and diagnosis, and further development of surgical instruments. Generalization of treatment outcomes should no longer apply given the varied results with disease stage. Confirmation of our tailored approach to management of the disease according to stage should soon be corroborated with an appropriate clinical trial. PMID- 14533899 TI - Vascular occlusion in the management of complicated multifetal pregnancies. AB - The decision to undergo ST is a personal one for the involved couple, and it can have many psychosocial implications. Appropriate counseling including offering all possible management options with related risks is imperative. Choosing the technique that best serves the clinical situation with minimization of maternal risks should be done taking under consideration the operator's experience. Ultimately, vascular occlusion techniques can help improve multifetal pregnancy outcomes in otherwise challenged gestations. PMID- 14533900 TI - Multifetal pregnancy reduction and selective termination. AB - MPR and ST are important options for patients who have multifetal pregnancies. Both procedures have been shown to be technically safe and result in acceptable pregnancy loss rates and GAs at delivery. An important caveat is that these findings are observed in centers that have vast experience performing this type of procedure and should not be generalized to all centers. The authors believe that the good outcomes reported here and elsewhere are a result of having a relatively limited number of operators adhering to a strict common protocol and that they should not be generalized to all centers. Awareness of the ethical and psychological issues aids counseling of patients and their follow-up, but more information is needed in this area. Finally, it is the authors' hope that advances in ART will decrease the need for MPR procedures in the future. PMID- 14533901 TI - A selective history of hernia surgery in the late eighteenth century: the treatises of Percivall Pott, Jean Louis Petit, D. August Gottlieb Richter, Don Antonio de Gimbernat, and Pieter Camper. AB - Of the different eras in the evolution of hernia surgery one of the most intriguing is the late eighteenth century, when surgeon/anatomists first began to publish their studies of the abdominal wall and the inguinal and femoral canals. It became known as the age of dissection, and many of the surgical successes of subsequent periods can be traced back to the anatomical knowledge gained from 1750 to 1800. These fifty years also served as the all-important transition era from text-only hernia treatises to lavishly illustrated monographs. The works of Percivall Pott, Jean Louis Petit, D. August Gottlieb Richter, Don Antonio de Gimbernat, and Pieter Camper were among the most influential hernia-related tomes of this time. Biographies of these five surgeons and extracts from their writings are presented in this article. PMID- 14533902 TI - Demographic and socioeconomic aspects of hernia repair in the United States in 2003. AB - Data from the National Center for Health Statistics reveals that approximately 800,000 groin hernia repairs were completed in the United States in 2003. More than 90% of these operations involve the use of mesh prosthesis and are performed on an outpatient basis. The two most common groin hernia repair techniques are the Lichtenstein and plug hernioplasties. Economic evaluation of groin hernia surgery demonstrates that the most important component of cost effectiveness is the aggregate time the patient spends in the operating room, recovery room, and the length of his or her overall stay in the facility. PMID- 14533903 TI - Classification systems for groin hernias. AB - All groin hernia classifications are somewhat arbitrary and artificial. Currently, there is no consensus among either general surgeons or hernia specialists as to a preferred system. A survey by Zollinger in 1998 of hernia specialists in North American and Europe showed, that although the Nyhus, Gilbert, and Schumpelick-Arit systems were commonly used, the majority of these specialists still used the traditional classification for groin hernias. It is apparent that only the traditional classification of groin hernias has stood the test of time. As stated by Fitzgibbons, "the primary purpose of a classification system for any disease is to stratify for severity so that reasonable comparisons can be made between various treatment strategies." Given the multiplicity of operative techniques and approaches for the repair of groin hernias, it appears that no one classification system can satisfy all. With time, it is likely that we surgeons will settle upon a given operation for a specific type of inguinal hernia. For that given operation to be accepted as proven best, however, it is essential the competing operations be applied to simliar (classified) groups of groin hernia patients. PMID- 14533904 TI - Anesthesia for groin hernia surgery. AB - Just as surgeons disagree about which is the best type of hernia repair, anesthesiologists disagree on which is the best anesthetic. Well-known and respected clinicians have used each of the techniques to produce anesthetics that, in terms of safety and dependable, speedy discharge, would have seemed impossible 15 years ago. I have mentioned problems, some more serious than others, with each of the techniques, but they are problems that can be addressed. All of the anesthetics can produce very satisfactory conditions and outcomes if carefully, thoughtfully, and gently applied. The best results, however, can be expected when a surgeon and anesthesiologist choose one technique, as we have done with epidural anesthesia, and develop a set of practices that enable them to exploit the benefits and minimize the problems inherent in that technique. PMID- 14533905 TI - The PerFix plug repair for groin hernias. AB - Since the mid-1990s, the PerFix plug hernioplasty has become one of the mainstays of a surgeon's operative armamentarium. The repair is a technically simple surgical operation, which can be used to treat most groin hernias. To demonstrate the simplicity and effectiveness of the plug technique, a 15-year experience with over 4400 patients is reported. Technical details are presented, including an in depth discussion concerning PerFix plug preparation and placement. PMID- 14533906 TI - The Lichtenstein repair for groin hernias. AB - The Lichtenstein tension-free mesh repair opened a new era in groin hernia repair. Without the risk of serious morbidity, it is now readily performed using local anesthesia in patients previously considered unsuitable for surgery. Fears of serious complications related to mesh implantation have proved to be without foundation. As a local anesthetic day care technique without the need for complex and expensive instrumentation, overall costs can be kept to a minimum without compromising the safety or the long-term success of the procedure. It is a straightforward technique to learn, with high patient acceptability and an exceedingly low recurrence rate. At present it must be considered the gold standard of groin hernia repair. PMID- 14533907 TI - The Kugel repair for groin hernias. AB - The Kugel repair is a minimally invasive but nonlaparoscopic preperitoneal hernia repair. It has certain advantages over other repairs, but will require a little additional effort initially to learn the repair and the associated anatomy encountered in this approach. This anatomy has not, historically, been well taught in medical schools. This repair is an attempt to achieve the fastest recovery possible after groin hernia surgery while assuring a very low risk of recurrence. PMID- 14533908 TI - Laparoscopic repair for groin hernias. AB - So where do things stand in 2003? Laparoscopic herniorrhaphy appears to result in less postoperative pain (acute and chronic) and in a shorter convalescence and an earlier return to work, compared with the open repair. It can be performed safely and with a low recurrence rate. However, it takes longer to do, is more difficult to learn, and costs more, all reasons why it is not more commonly performed. Currently, laparoscopic herniorrhaphy accounts for 15% to 20% of hernia operations in America and around the world. Who can blame the surgeon in a community practice for opting for the open mesh repair, operating on familiar anatomy, and using familiar techniques? Nevertheless, with efforts to cut costs by eliminating disposable equipment and honing skills to decrease operating time, laparoscopic herniorrhaphy will probably continue to be a contender, especially for the younger patient who wants to return to work quickly and for patients with bilateral and recurrent hernias. It is arguable that surgeons should possess skill in both open and laparoscopic techniques and should know the indications for each--some hernias are best repaired laparoscopically. That said, laparoscopic herniorrhaphy will most likely be performed by those with a special interest and proficiency in the technique. At the least, the laparoscopic revolution and laparoscopic hernia repair have helped elevate the study of hernia anatomy and herniorrhaphy to a position it deserves and this has made us all better hernia surgeons. What was once the stepchild of general surgery now occupies a more prominent and respectable place. With the continuing efforts of dedicated, energetic investigators, we should continue to see advances in the safe and effective repair of this most common of surgical maladies. PMID- 14533909 TI - The Shouldice repair for groin hernias. AB - The Shouldice pure tissue repair for groin hernias, a tested and proven procedure, continues to provide great patient satisfaction. The complete dissection lends itself to the discovery of secondary hernias. The procedure is designed so that these secondary defects are easily incorporated into the repair. Mesh is employed infrequently, only in those cases where there is a lack of tissue or scarring has minimized flexibility. Performed under local anesthetic, complications are minimal and early return to work is a feature. With a more than 50-year history of thousands of successful operations annually, the Shouldice procedure should continue to merit consideration by all surgeons striving for excellence in hernia repair. PMID- 14533910 TI - Femoral hernia repair. AB - Femoral hernia repair has a long history. In the nineteenth century, simple closure of the femoral orifice by the femoral approach was favored. Such renowned surgeons as Bassini, Marcy, and Cushing authored papers about the femoral approach to femoral hernia. The recurrence rate was so high, however, that it was replaced by the inguinal approach. The man who popularized the inguinal approach was Chester McVay, who demonstrated the precise insertion of the tranversus abdominis muscle and transversalis fascia to the Cooper's ligament. He used Cooper's ligament for the femoral hernia repair by the inguinal approach. The complication and recurrence rate after the Cooper's ligament repair for femoral hernia was not satisfactory, however, due to tension on the approximated tissues, which caused postoperative pain and inability to resume normal activities. Irving Lichtenstein first introduced the plug technique to femoral hernia repair and it was further developed by Gilbert and Rutkow. In the present series, all elective cases were repaired by the PerFix mesh plug technique without any complications. Patients were discharged from the hospital on the first postoperative day and returned to normal activities shortly thereafter. These patients had few complaints of pain in the groin. The operating time using a PerFix plug was markedly shorter when contrasted with the Cooper's ligament repair. No infection of the prosthesis occurred, even in the cases in which the small intestine was necrotic and resected. From our 7-year experience of mesh plug femoral hernia repairs, I have come to regard this operation as the first choice in elective and noninfected cases of femoral hernia. In strangulated cases in which severe infection occurs. Cooper's ligament repair should be used, because there is a risk or infection to implanted prosthesis. Finally, femoral hernia is usually thought of as requiring emergency surgical treatment. Only 30% of our cases were treated as emergency operations, however, whereas 70% were elective. Unless patients complain of severe abdominal pain or ileus, surgeons need not perform emergency operations. In summary, the PerFix mesh plug hernia repair for femoral hernia has resulted in a reduced recurrence rate, shortened hospital stay, and a low rate of postoperative complications. PMID- 14533911 TI - Umbilical and epigastric hernia repair. AB - The repair of umbilical and epigastric hernias still represents a challenge to surgeons. Although a common and relatively simple procedure, there is no exact protocol today on how the repair should be done. The Mayo technique and its alterations could not stand the test of time: a recurrence rate of 20% and higher is not acceptable for any surgical procedure. Although there is no consensus opinion, one thing is clear: the importance of an anatomic repair without tension and without an artificial enlargement of the defect. In 1987 Lichtenstein reported on 6321 cases of herniorraphy with a tension free repair, and in 1994 Stuart reemphasized that special importance in his editorial in the Lancet. A newer study from Brancato and coworkers in Italy also states the advantage of a tension-free prosthetic repair in 16 patients with epigastric hernia. We have gone even further and recommend a tailored-to-the-patient repair using a customized polypropylene mesh and a one-layer running suture. The advantages should be obvious: no artificial creation of an even bigger than original defect, a completely tension-free repair, and little to no recurrence of the hernia. Our results clearly prove that assumption. Moreover, the procedure is extremely safe and complications are very rare and minor. We conclude that using a mesh plug in a customized tension-free repair of umbilical and epigastric hernia shows many advantages over the commonly used methods. And we finally conclude with the words of Albert Einstein: "The only source of knowledge is experience." PMID- 14533912 TI - Incisional hernia repair. AB - Incisional ventral hernias are a common problem encountered by surgeons, with over 100,000 repairs being performed annually in the United States. Although many predisposing factors for incisional ventral hernia are patient-related, some factors such as type of primary closure and materials used may reduce the overall incidence of incisional ventral hernia. With the advent of prosthetic meshes being used for incisional ventral hernia repair, the recurrence rate has dropped to approximately 10%. More recently, with the development of prosthetic mesh that is now safe to place intraperitoneally, the recurrence rate has dropped to under 5%. The current controversies that exist for incisional ventral hernia repair are which approach to use (open versus laparoscopic) and what type of fixation (partial- versus full-thickness abdominal muscular/fascial wall) is necessary to stabilize the position of the mesh while tissue ingrowth occurs. During the next decade the answers to these controversies should be available in the surgical literature. PMID- 14533913 TI - Spigelian and other uncommon hernia repairs. AB - This article offers an overview of abdominal wall hernias, which are uncommon because of the unusual contents of their sacs. These include: Meckel's diverticulum, segments of the intestinal wall antimesenteric border, the vermiform appendix, the bladder; plus the penetration by the sac within the different muscle layers of the abdominal wall. These hernias present diagnostic difficulties and some are associated with high morbidity, but modern technology may help their prognosis. This article describes their histories and their unique presenting signs and symptoms and suggests up-to-date methods of imaging, as well as surgical tactics and technique. PMID- 14533914 TI - Complications of open groin hernia repairs. AB - As large numbers of patients undergo hernioplasty each year the surgical technique should be a simple one. The results obtained by general surgeons using various open, tension-free techniques, irrespective of the anesthetic used, are excellent and appear to approach those of specialists. This can not be said for laparoscopic hernioplasty, which has a well-known learning curve, is more expensive and is not without complications, some of which may be serious or life threatening. Although proper training in laparoscopic techniques is essential, the same applies to open repair with mesh. Surgical residents should be taught open-mesh repairs under local anesthesia before embarking on training in laparoscopic techniques if they show interest in pursuing this approach. Indeed, we may soon be faced with an increasing number of patients who are not fit for a general anesthetic. Not all hernias need be repaired by specialists and visiting centers with experience in the use of different prosthetic devices allows you to draw your own conclusions. Finally, when consulting a patient with an inguinal hernia, primary or recurrent, the surgeon should pose the question "which combination of anesthetic and hernia repair is the safest and best for my patient?" Local anesthesia with appropriate analgesia and sedation is the safest of all techniques and is suitable for most if not all open repairs. Using this approach, any type of open-mesh repair makes the ideal combination and all can be safely carried out on an ambulatory basis. Attention to surgical technique is paramount, and given the number of hernias repaired annually, it is pertinent to recall the words of Wakely, who said "A surgeon can do more for the community by operating on hernia cases, and seeing that his recurrence rate is lower, than he can by operating on cases of malignant disease." PMID- 14533915 TI - Outcomes analysis for groin hernia repairs. AB - Outcomes studies after inguinal hernia operations show a pattern of continued improvement throughout the last decade. OHRs are intrinsically less costly and less complicated to perform than LHR. The most recent modification in OHR has been the improvement of the mesh prostheses for insertion into the preperitoneal space. The earlier return to work seen in the early 1990s with LHR has been offset by comparable recuperation in the late 1990s associated with improvements in prosthetic repair in OHR. Both behavioral and technical factors must be evaluated to improve outcomes with hernia surgery. PMID- 14533916 TI - Teratogenic effect of broad-band electromagnetic field on neonatal mice (Mus musculus). AB - Pregnant mice (Mus musculus), strain Swiss Webster, were exposed to a continuous electromagnetic field (12.8V/m) beginning in the third week of pregnancy. Histological and hematological analysis showed gender specific responses in 21 day-old mice after in-utero and post-natal continuous exposure. Automated lymphocyte percentage and total white blood cell counts were significantly elevated in exposed 21 day-old female mice compared to control mice. Lymphoma like cells were seen in higher numbers in exposed 21 day-old male mice. Megaloblastic changes, such as hypersegmented neutrophils, were observed in exposed mice. The blood from control neonatal mice was more viscous than that of exposed mice, enough to interfere with making a blood smear. The adult female mice showed no significant differences in the above hematologic parameters between exposed and control groups. Histological study showed the following pathological changes in the adrenal cortex: degeneration/necrosis in the zona glomerulosa; hypertrophy in zona reticularis; degeneration/necrosis, intracytoplasmic inclusions and inflammation in the zona fasciculata/reticularis, more prominent in exposed female neonates; and lipidosis in the zona fasciculata. In the adrenal medulla: atrophy was more common in exposed female neonates; and intracytoplasmic inclusions and vacuolation were more common in exposed male neonates. Cystic proliferations were found in the cortical area of the thymus. In the medulla of the thymus, there was vacuolation, inflammation, or eosinophilic intracytoplasmic inclusions in exposed adults. Behavioral differences occurred in both neonates and adult females. Control neonates were able to manipulate through a maze more quickly than exposed neonates; and control adult females displayed more thorough grooming behavior than exposed mothers, and maintained more distance between the nest and dropping location than did the exposed group. PMID- 14533917 TI - Ammonium removal using a vertically moving biofilm system. AB - Ammonium-nitrogen (NH4-N) removal using a vertically moving biofilm system was investigated at a temperature of 11 degrees C. The biofilms in this process were grown on biofilm modules, consisting of high surface area plastic media, which were vertically and repeatedly moved up into the air and down into the wastewater. The maximum NH4-N removal efficiency was up to 98.8% and the areal NH4-N removal rates, based on the surface area of the biofilm modules, were within the range of 0.68-1.94 g NH4-N m(-)2 day(-1). The kinetics of ammonium removal occurring after carbonaceous oxidation was completed can be simulated by using a zero-order empirical model. The ammonium removal rate was found to be proportional to the square root of the mass of biofilms on the modules. Monitoring DO concentrations in the bulk fluid showed that the vertically moving biofilm system possessed good aeration efficiency. PMID- 14533918 TI - A preliminary study of the application of some predictive modeling techniques to assess atmospheric mercury emissions from terrestrial surfaces. AB - Predictive modeling techniques are applied to investigate their potential usefulness in providing first order estimates on atmospheric emission flux of gaseous soil mercury and in identifying those parameters most critical in controlling such emissions. Predicted data by simulation and statistical techniques are compared to previously published observational data. Results showed that simulation techniques using air/soil coupling may provide a plausible description of mercury flux trends with a RMSE of 24.4ngm(-2)h(-1) and a mean absolute error of 10.2ngm(-2)h(-1) or 11.9%. From the statistical models, two linear models showed the lowest predictive abilities (R2=0.76 and 0.84, respectively) while the Generalized Additive model showed the closest agreement between estimated and observational data (R2=0.93). Predicted values from a Neural Network model and the Locally Weighted Smoother model showed also very good agreement to measured values of mercury flux (R2=0.92). A Regression Tree model demonstrated also a satisfactory predictability with a value of R2=0.90. Sensitivities and statistical analyses showed that surface soil mercury concentrations, solar radiation and, to a lesser degree, temperature are important parameters in predicting airborne Hg flux from terrestrial soils. These findings are compatible with results from recent experimental studies. Considering the uncertainties associated with mercury cycling and natural emissions, it is concluded, that predictions based on simple modeling techniques seem quite appropriate at present; they can be useful tools in evaluating the role of terrestrial emission sources as part of mercury modeling in local and regional airsheds. PMID- 14533919 TI - Ambient concentrations of total suspended particulate matter and its elemental constituents at the wider area of the mining facilities of TVX Hellas in Chalkidiki, Greece. AB - To assess ambient air quality at the wider area of TVX Hellas mining facilities, the Total Suspended Particulate matter (TSP) and its content in characteristic elements, i.e., As, Cd, Cu, Fe, Mn, Pb, Zn are being monitored for more than thirty months as part of the established Environmental Monitoring Program. High Volume air samplers equipped with Tissue Quartz filters were employed for the collection of TSP. Analyses were effected after digestion of the suspended particulate with an HNO3-HCl solution and determination of elemental concentrations with an Atomic Absorption Spectroscopy equipped with graphite furnace. The sampling stations were selected to record representatively the existing ambient air quality in the vicinity of the facilities and at remote sites not affected from industrial activities. Monitoring data indicated that the background TSP concentrations ranged from 5-60 microg/m3. Recorded TSP concentrations at the residential sites close to the facilities ranged between 20 100 microg/m3, indicating only a minimal influence from the mining and milling activities. Similar spatial variation was observed for the TSP constituents and specifically for Pb and Zn. To validate the monitoring procedures, a parallel sampling campaign took place with different High Volume samplers at days where low TSP concentrations were expected. The satisfactory agreement (+/- 11%) at low concentrations (50-100 microg/m3) clearly supported the reproducibility of the techniques employed specifically at the critical range of lower concentrations. PMID- 14533920 TI - VOC emission characteristics of petrochemical wastewater treatment facilities in southern Taiwan. AB - The wastewater treatment plants (WWTPs) of three representative petrochemical plants in southern Taiwan were sampled to investigate their VOC emission characteristics. Generally, emissions decline along the process flow, while the VOC concentration accumulates to as much as around 2400mg/m3 (as total hydrocarbons) at several closed vessel vents during the primary treatment. VOC emission rates (g/s) and fluxes (g/m2 hr) exhibit a similar trend of reducing concentrations in the WWTPs. From the field analysis data. VOC emission potential is best indicated by the concept of emission flux rather than concentration or emission rate. The Water 8 emission model version 4, developed by the U.S. EPA in 1995, was used to simulate the emission rate. The output results were compared to the calculated VOC liquid-gas mass equilibrium based essentially on the saturated gas concentration (Cg*) of the individual VOC components in wastewater. The comparisons clearly identified the proposed mass transfer approach using (Cg* as a reliable timesaving alternative to Water 8, which requires numerous input parameters and water analysis data. PMID- 14533921 TI - Odor control in evaporation ponds treating olive mill wastewater through the use of Ca(OH)2. AB - Different amounts of Ca(OH)2 were added in 2 L beakers containing 1 L of olive mill wastewater (OMW). The mixture was stirred for 45 min and left to settle. Wastewater analysis was used in order to determine the effect of the different amounts of calcium hydroxide in the treating process, three days after the application. The Odor Detection Threshold was used for determining the effect of the treatment in the odors produced in the beakers, three and 30 days after. Both sets of measurements indicated an important reduction in wastewater pollutants and odor emission when 10 g/L of Ca(OH)2 were added. In order to evaluate these results in more realistic conditions. 10 L plastic containers were filled with 6 L of OMW, relevant amounts of Ca(OH)2 were added, the mixture was stirred manually and left to settle in the open. Again, 10 g/L of calcium hydroxide produced the best results in odor reduction and wastewater treatment. PMID- 14533922 TI - Estimation of methane emission from rice cultivation in Korea. AB - This study focused on estimating a methane (CH4) inventory and developing mitigation options in South Korea, and was performed jointly in an integrated national research program on CH4 from rice fields conducted by three National Agricultural Research Institutes, under the Rural Development Administration during 1993-1997. Methane emissions were measured by a closed chamber method, in rice plots at three locations (Suwon, Iksan, and Milyang) with the single rice cropping system. All experimental data from 5 years of study were summarized and used for calculating nation-wide CH4 emissions. Temperature, soil type, cultural practices, water management, organic matter management, and cultivar selection significantly affected the fluctuations of CH4 emissions. The two most promising mitigation options for reducing emissions were altered water management, in particular mid-season aeration by short-term drainage, and improved organic matter management, by promotion of aerobic degradation through composing or soil incorporation. Annual total CH4 emission in Korea changed from 410Gg in 1990 to 339Gg in 2000, due mainly to a decrease in the cultivated area of paddy rice. If we convert annual CH4 emission to Global Warming Potential as CO2 equivalent, it amounts to 7.1 M CO2 t yr(-1) of greenhouse gas emitted to the atmosphere in 2000 from the rice fields in Korea, which is just 5.3% of the annual CO2 emission from the industry and energy sector. More importantly, the balance between CO2 uptake by photosynthesis and CO2 emission is positive (a net sink), so that rice culture actually has net benefits for the global atmospheric carbon issue. Further reductions in emission amounts, by following recommendations in this article, could make these benefits even greater. PMID- 14533923 TI - Natural occurrence of arsenic in shallow groundwater, Shanyin, Datong Basin, China. AB - In Shanyin, Shanxi province, China, As concentration in shallow groundwater exceeds guide concentrations, set internationally and nationally at 10-50 microg/L, and may reach 1932.0 microg/L, which has resulted in severe clinical symptoms of arsenic toxity. In this article, chemical characteristics of groundwaters containing anomalous As and mechanisms of arsenic release to shallow groundwaters were studied. Groundwaters containing anomalous As in the study area were characterized by higher pH, higher concentration of phosphate, higher concentration of naphthenic acid, and lower concentrations of sulfate and nitrate. Microbial metabolism of sedimentary organic matter, which is present as high as 1.0% organic C, results in the lower concentrations of sulfate and nitrate. The reactions decrease Eh and produce CO2, which promotes the dissolution of carbonates and increases pH. The clay minerals and colloids including organic matters scavenging arsenic would release arsenic to groundwater in higher pH and lower Eh environment. In addition, the competitive absorption between As and anions (such as phosphate and fluoride) contributes to the release of As from kaolinite, montmorillonite, illite, and Fe oxyhydroxides. Naphthenic acid also promotes mobilization and translocation of As in groundwater systems. PMID- 14533924 TI - Photocatalytic degradation of two selected textile dye derivatives, eosine yellowish and p-rosaniline, in aqueous suspensions of titanium dioxide. AB - Thephotocatalytic degradation of two selected textile dye derivatives, eosine yellowish (1) and p-rosaniline (2) has been investigated in aqueous suspensions of titanium dioxide under a variety of conditions. The degradation was studied by monitoring the change in substrate concentration employing UV spectroscopic technique and decrease in Total Organic Carbon (TOC) content as a function of irradiation time under a variety of conditions. The degradation of the dye was studied under different conditions such as pH, catalyst concentration, substrate concentration, different types of TiO2 and in the presence of electron acceptor such as hydrogen peroxide (H2O2), potassium bromate (KBrO3), and ammonium persulphate (NH4)2S2O8) besides molecular oxygen. The degradation rates were found to be strongly influenced by all the above parameters. The photocatalyst Degussa P25 was found to be more efficient as compared with other photocatalyst in the case of p-rosaniline, whereas UV100 was found to be better for the degradation of eosine yellowish. The xanthene dye (1) was found to degrade faster as compared to the triphenylmethane dye (2). The degradation products were analyzed by GC/MS technique and plausible mechanism for the formation of products have been proposed. PMID- 14533925 TI - Photochemical degradation performance of quinoline aqueous solution in the presence of hydrogen peroxide. AB - Photochemical degradation performance of quinoline aqueous solution in the presence of H2O2 was carried out, and some intermediates produced during quinoline degradation were also identified tentatively. The experimental results showed that the advanced oxidation of quinoline by UV/H2O2 process accorded well with the pseudo first order kinetics, and the dependence of concentrations of H2O2 and quinoline, and pH value on the photodegradation kinetics has been investigated in detail. It is found that the concentrations of hydrogen peroxide and quinoline have opposite effect on photodegradation kinetics. That means the photodegradation rate of quinoline increased significantly as the hydrogen peroxide concentration increasing, while the photodegradation rate decreased critically as the initial concentration of quinoline increasing. It also concluded that the photodegradation of quinoline by H2O2/UV process is more favorable under alkali solution than acid solution. PMID- 14533926 TI - Concentration of polycyclic aromatic hydrocarbons (PAHs) in moss (Hypnum cupressiforme) from Hungary. AB - Mosses of the species Hypnum cupressiforme were collected from different parts of Hungary to investigate the air quality of the region. The concentrations of polycyclic aromatic hydrocarbons (PAHs) in species were determined. Low molecular weight PAHs (up to three ring compounds) contribute more than 99% to the concentrations of PAHs in Moss samples. The pattern of PAHs suggests local sources rather than long-range transport of contamination in the air. Possible local sources are industrial, urban, and traffic activities. The results indicate incomplete combustion of fuel is largely responsible for the air quality of the area. PMID- 14533927 TI - Removal of reactive dye from wastewater by adsorption using ECH cross-linked chitosan beads as medium. AB - The feasibility of the removal of reactive dye from wastewater using a novel adsorbent, ECH cross-linked chitosan beads, as medium was examined. The effect of the pH and the initial concentration of the dye (RR222) solution on the adsorption capacity of the ECH cross-linked chitosan beads were also investigated. It was found that the initial dye concentration and the pH of the solution significantly affected the adsorption capacity. An increase in initial dye concentration results in the increase of adsorption capacity while an acidic pH was found to be favorable for the adsorption of dye. It was also found that the equilibrium adsorption of RR222 could be adequately described by using the Langmuir model (r2 > or = 0.999). Moreover, results showed that the adsorption rate of RR222 onto ECH cross-linked chitosan beads could be described by using the second-order kinetic model, suggesting that chemical sorption instead of mass transfer was the rate-limiting step for the adsorption process. The maximum monolayer adsorption capacity obtained from the Langmuir model was extremely high as compared to the data reported in literature; 2252 g/kg at 30 degrees C with a pH of 3.0. Therefore, ECH cross-linked chitosan beads could be a feasible medium for the removal of reactive dye from wastewater and potentially an alternative for the decolorization of wastewater. PMID- 14533928 TI - Anaerobic-aerobic sewage treatment using the combination UASB-SBR activated sludge. AB - The performance of a pilot scale sewage treatment system composed of an upflow anaerobic sludge blanket (UASB) digester and a sequencing batch activated sludge reactor (SBR) is described. The system constitutes a simple, economic, and compact treatment option and is able to produce consistently a final effluent quality that is equal or better than that of a conventional activated sludge plant. The entire treatment system had a retention time of only 9h and an average operational temperature of 25 degrees C. Due to the efficient anaerobic pre treatment the aerobic sludge production was low and could easily be accommodated for stabilisation in the UASB reactor. The system exhibited excellent operational stability with full nitrification for aerobic sludge ages longer than 9 days. At shorter sludge ages there was excessive wash out of sludge particles and the sludge mass could not be maintained in the SBR reactor. Sludge settleability was good throughout the experimental investigation period of one year. The reduction of the reactor volume and oxygen consumption was more than 50% compared to conventional activated sludge. The anaerobic excess sludge had a high concentration and good stability so that its dewatering and final disposal was a relatively minor problem. PMID- 14533929 TI - Concentration of copper, iron, zinc, cadmium, lead, and nickel in boar semen and relation to the spermatozoa quality. AB - The concentration of copper, iron, zinc, cadmium, lead, and nickel as well as its relation to spermatozoa quality was investigated. The semen samples were analyzed by atomic absorption spectrophotometry (AAS). The concentration of copper in boar semen was 1.64 +/- 0.28 mg kg(-1) and of iron 16.14 +/- 10.35 mg kg(-1). The concentration of zinc in boar semen reached an average value of 171.74 +/- 64.72 mg kg(-1) and the level of cadmium reached 0.01-0.16 mg kg(-1) with the average value of 0.05 mg kg(-1). The analysis of lead showed that the concentration of this element in boar semen was 0.02 +/- 0.03 mg kg(-1) and the average level of nickel was 0.06 +/- 0.08 mg kg(-1). The total percentage of pathological spermatozoa was 9.82 +/- 1.47%. Detail analysis determined 3.18% of separated flagellum, 2.26% knob twisted flagellum, 0.88% flagellum torso, 0.85% flagellum ball, 0.42% broken flagellum, 0.23% retention of the cytoplasmic drop, 0.14% small heads, 0.03% large heads, and 1.83% forms other of pathological changes. Correlation analysis showed significant (p < 0.05) positive correlation between copper and lead (r = 0.52). High correlation between small head and knob twisted tail (r = 0.67), small head and broken flagellum (r = 0.88) as well as between small head and total number of pathological spermatozoa (r = 0.73) was determined. PMID- 14533930 TI - Extraction of heavy metals by amines adsorbed onto silica gel. AB - Previous work has established the possibility of using known coordinating agents supported on silica gel in the removal of heavy metals such as lead, copper, cadmium, and nickel from aqueous media. Functionalized silica gel has been used in the removal of heavy metals with notable success. Silica gel is currently being used as a support for various monofunctional aliphatic amines as coordinating ligands. The current study reports the results of an investigation involving the use of saturated, straight chain primary amines as coordinating ligands in the removal of copper(II), cadmium(II), lead(II), nickel(II), and silver(I) ions from aqueous solutions of known concentration. Primary amines used in this investigation were n-butylamine, n-hexylamine, n-octylamine, n decylamine, and n-hexadecylamine. PMID- 14533931 TI - The feasibility study of the octane-enhancing process: isobutylene dimerization using C4 mixture. AB - The dimerization of isobutylene to diisobutylene in C4 mixture was evaluated using various catalysts such as nickel salts system, zeolite system and acidic ion exchange resin. There were shown that the strong acidic type ion exchange resin (Amberlyst 15) under 6 kg/cm2 and 60 degrees C gave rise to 90% conversion and more than 60% selectivity to diisobutylene with acceptable amounts of oligomer as a best catalyst. Therefore, Amberlyst 15 was employed to investigate the optimum reaction condition. In the batch reaction, since the catalyst activity was increased with the increasing reaction temperature, the reaction time should be shortened appropriately to prevent a high yield of oligomer. The reaction rate was increased by the increased amount of catalyst without much change in the product composition. The formation of oligomer was inhibited when a small amount of organic solvents (methyl tert-butyl ether) are attended. In the fixed-bed study, the results indicate that the increase in reaction temperature was accompanied by the increase in reaction rate, however, the selectivity to the dimer was affected. Higher WHSV will enhance the selectivity but reduce the conversion. Conclusively, the appropriate reaction condition was 45 degrees C with WHSV of 10. PMID- 14533932 TI - Mineral accumulation, growth, and physiological functions in Dalbergia sissoo seedlings irrigated with different effluents. AB - Present study aimed to observe the mineral accumulation added through effluent application and their influence on physiological functions and growth of Dalbergia sissoo and ultimately to find out suitable combination of industrial and municipal effluent for their utilization in raising tree plantation. Dalbergia sissoo seedlings were irrigated with: canal water (T1); municipal effluent (T2); textile effluent (T3); steel effluent (T4); textile + municipal effluent in 1:1 ratio (T5); steel + municipal effluent in 1:2 ratio (T6); steel + municipal + textile in 1:2:2 ratio (T7); and steel + textile in 1:2 ratio (T8). Mineral accumulation, water relations and gas exchange, growth and biomass production were the recorded observations. Mortality occurred within a day for the seedlings in T4, 45 days in T6, and 60 days in T7 and T8 treatments. This was probably the result of high (P<0.01) Mn, Fe, Cu, and Zn and low N, P, K, Ca, and Mg concentration in different parts of the seedlings in these treatments affecting physiology and growth. Reduction in rate of photosynthesis (Pn) and transpiration (T) to the level of 90%, leaf water potential (LWP) and increased stomatal resistance (R) at two months of age is the indicator of metal toxicity in these treatments. Pn and T were 20 and 17% high in T2 treatment, respectively. However, the reduction was only 21 and 10% in Pn and 18 and 26%, respectively in the seedlings of T3 and T5 than that in T1 treatment. During nine months of observation, the seedlings of T2 and T5 maintained high LWP and same rate of Pn and T but they reduced significantly (P<0.01) in the seedlings of T3 treatment affecting biomass production. This was believed to be due to increase in Na concentration leading to decrease in Mg, Ca, Mn, Fe, Cu and Zn concentration and ratio of Mg/Na and Mg/K and increased ratio of N/Mg and K/Ca + Mg. The enhanced physiological functions in T5 treatment were probably the result of ameliorative effect of municipal effluent through increased mineral status producing biomass equivalent to that in T1 treatment at 10 months of age. The seedlings of T2 attained greater (P<0.01) height and collar diameter and produced 120g seedling( 1) of dry biomass. The study suggests that addition of excess mineral elements leads to metal toxicity that adversely affects physiology and ultimately growth and productivity of tree seedlings. Mixing of effluents to ameliorate the toxic effects could be the better management practices for their use in tree irrigation. PMID- 14533933 TI - Effects of soil fines and surfactant sorption on contaminant reduction of coarse fractions during soil washing. AB - The reduction of contaminants sorbed on the coarse fraction of soils to the level below clean-up requirements is essential for an effective soil washing process. This study investigated the effects of soil texture and surfactant sorption on the reduction of total petroleum hydrocarbons (TPH) in coarse fraction during soil washing. Batch TPH sorption experiments were conducted on soil slurry with various soil fine/coarse ratios and surfactants Octylpheny polyoxyethylene (TX 100) and Dodecylpyridinium chloride (DPC) at the dosage below their saturation levels of sorption. In a sandy loam soil of low silt and clay contents, increasing the fine/coarse ratio from 0.4 to 1.2 without adding surfactants resulted in a reduction of TPH levels in the coarse fraction by 30%. Increasing the fine/coarse ratio along with sorbed surfactant (3000 mg TX-100 or 10,000 mg DPC per kg soil) further reduced TPH concentrations in the coarse fraction. For a silty loam soil already containing a high percentage of fine particles, increasing the fine/coarse ratio from 5.9 to 18.8 without surfactant addition yielded no further TPH reduction in the coarse fraction. On the other hand, surfactant sorption at the fine/coarse ratio of 5.9 improved the washing efficiency of the coarse fraction. These experimental results suggested the importance of high contents of soil fines and surfactant sorption in achieving low contaminant concentrations of coarse fractions during soil washing. PMID- 14533934 TI - A comparison of water quality indices for coastal water. AB - The present article discusses and compares five different water quality indices, viz arithmetic water quality index, multiplicative water quality index, unweighted arithmetic water quality index, unweighted multiplicative water quality index, and Harkin's water quality index, which were considered for characterizing the coastal water quality at the Jawaharlal Nehru Port Trust, Bombay, India. Dissolved oxygen, pH, biochemical oxygen demand (BOD), temperature, suspended solid, and turbidity were used as the parameters of water quality indices. The value function graphs used for above-mentioned variables were developed using harbour water quality standards and aquatic life. The product moment correlation coefficients for various water quality indices were determined using the SPSS software package to evaluate correlation among various indices. It was found that the unweighted arithmetic water quality index was higher than weighted arithmetic water quality index while the multiplicative water quality index was lower than unweighted multiplicative water quality index. All the indices were well correlated with each other except Harkin's water quality index. The Harkin's water quality index was different from other water quality indices. The comparison of different form of indices showed that the multiplicative water quality index was the most suitable water quality index for coastal waters. PMID- 14533935 TI - Neuroendocrine tissue-specific transcription factor, BETA2/NeuroD, in gastric carcinomas: a comparison with chromogranin A and synaptophysin expressions. AB - BETA2/NeuroD (NeuroD) is a basic helix-loop-helix type of transcription factor mainly involved in neuroendocrine differentiation. In this study, we evaluated the prevalence of neuroendocrine differentiation in gastric carcinomas by analyzing the NeuroD expression in comparison with those of chromogranin A and synaptophysin. Of the 70 cases of gastric adenocarcinoma, the expressions of NeuroD, chromogranin A, and synaptophysin were detected in 17 (24.3%), four (5.7%), and 24 cases (34.3%), respectively, with preferential expressions in a non-solid type of poorly differentiated adenocarcinoma. The expression pattern of NeuroD was mostly concordant with that of synaptophysin and partly with chromogranin A, indicating that NeuroD serves as a good neuroendocrine marker in gastric adenocarcinomas. On the other hand, no immunoreactivity against NeuroD was detectable in nine cases of gastric neuroendocrine carcinomas, including small cell carcinomas, despite the presence of synaptophysin and chromogranin A expressions. These findings led us to conclude that neuroendocrine differentiation is estimated to be present in 20-30% of gastric adenocarcinomas with preference to a non-solid type of poorly differentiated adenocarcinoma. In addition, the negative expression of NeuroD in neuroendocrine carcinomas suggests that other regulatory mechanisms are possibly involved in the development of neuroendocrine carcinoma. PMID- 14533936 TI - Vascular remodeling process in reversibility of pulmonary arterial hypertension secondary to congenital heart disease. AB - Pulmonary vascular remodeling process was analyzed using morphometry in lung biopsy specimens taken from 26 children, aged 6 to 160 months, who had congenital heart disease and significant pulmonary arterial hypertension. Reparative surgery was performed in 22 patients and palliative surgery was performed in four patients. One patient expired postoperatively and four others after hospital discharge. Vascular remodeling examination revealed a characteristic pathological picture: pronounced medial thickening with increased collagen content (fibrosis), without significant arterial intimal proliferation. At a mean follow-up of 44 months, 72% of the survivors were asymptomatic with no medication. Diagnosed by echocardiogram, 22% of these patients were shown to have pulmonary arterial hypertension. The characteristic pathological features described above occurred in 38% of the patients who either expired or had pulmonary hypertension postoperatively. These findings were an aid to identifying a high risk group in which the outcome does not meet expectations for the classical grade I and II changes. We concluded that the presence of isolated medial thickening does not ensure either survival or a normal postoperative pulmonary arterial pressure at late follow-up and that the collagen content can be a better reference for good outcome. Early intracardiac repair is recommended before the development of significant medial fibrosis. PMID- 14533937 TI - Cdc2 expression in primary thyroid lymphoma: its relationship with biological aggressiveness and G2 cyclins. AB - Previous studies have demonstrated that the G2-M modulators contribute to the progression of human neoplasms. In this study, we investigated the expression of these modulators, cyclin-dependent kinase 1 (cdc2), and G2 cyclins, cyclin A and cyclin B1, in primary thyroid lymphoma. Cdc2 immunoexpression was observed in 51.0% of the 49 cases examined and was related to grade of malignancy, high Ki-67 labeling index, and aberrant p53 expression. The incidences of immunoexpression of cyclin A and cyclin B1 were 63.3% and 40.9%, respectively, and they were also related to the above three parameters. Furthermore, a correlation was found between the immunoexpression of cdc2 and G2 cyclins. These findings suggest that the cdc2 and G2 cyclins play an important role in the progression of thyroid malignant lymphoma. PMID- 14533938 TI - Immunophenotyping of angioimmunoblastic T-cell lymphomas by multiparameter flow cytometry. AB - Angioimmunoblastic T-cell lymphoma (AITL) is a distinct form of peripheral T-cell lymphoma (PTCL) frequently involving lymph nodes, spleen and bone marrow, and is associated with systemic symptoms. Its histologic features may be subtle at an early phase and difficult to diagnose. Despite the success of flow cytometry (FCM) in diagnosing B-cell neoplasm, FCM has not been widely accepted as a useful method for establishing the diagnosis of PTCL. Recently, the neoplastic T-cells in AITL have been shown to express CD10. We prospectively applied multiparameter FCM immunophenotyping to three cases of histologically confirmed AITL and identified a small (5-7%) population of CD4+/CD10+ T-cells in two cases. In one case, the CD4+/CD10+ population lacked surface signals of CD3 and CD7, but strongly expressed CD2, whereas CD45 expression was very weak; partial loss of surface CD3 was observed in the other. None of the lymph nodes with reactive hyperplasia, B-cell lymphomas, or Hodgkin's lymphoma studied during the same time period contained the CD4+/CD10+ population. These findings suggest that addition of CD4/CD10 and CD3/CD10 to FCM immunophenotyping panels is useful in the diagnosis of AITL. To the best of our knowledge, this is the first report to demonstrate CD10-expressing T-cells in AITL by FCM. PMID- 14533939 TI - Classical Hodgkin lymphoma occurring in clusters of nodal marginal zone B lymphocytes in association with progressive transformation of germinal center. A case report. AB - We present a case of a classical Hodgkin lymphoma occurring in clusters of marginal zone B-lymphocytes (MZBLs). Most lymphoid follicles possessed hyperplastic germinal centers, while a portion of the follicles exhibited a progressive transformation of the germinal center (PTGC). Clusters of MZBLs showed a perifollicular distribution. The classic Reed-Sternberg cells were found in clusters of MZBLs. A portion of the Reed-Sternberg cells were CD15+, CD20+, CD30+, CD79a+, fascin+, vimentin+, EMA-, and bcl-2-. Some Reed-Sternberg cells were surrounded by CD3+ CD45RO+ CD57-rosettes. In situ hybridization studies demonstrated strong expression of EBER in classic Reed-Sternberg cells and their variants. The overall morphological, immunohistological, and EBV findings confirmed that the present case is a classical Hodgkin lymphoma. The MZBLs were CD20+, CD79a+, sIgM+/-, sIgD-, CD5-, CD21-, CD43-, CD45RO-, and Bcl-2-. Some MZBLs had polytypic intracytoplasmic immunoglobulin. Problems arising in the differential diagnosis between lymphocyte-predominant Hodgkin lymphoma and PTGC have been described. An occasional association between MZBLs clusters and PTGC has been reported previously. This case suggests that classical Hodgkin lymphoma should be added to the differential diagnosis of PTGC. PMID- 14533940 TI - Small cell carcinoma of the endometrium: report of a case with analysis of Wnt/beta-catenin pathway. AB - Small cell carcinoma of the endometrium (SCCE) is extremely rare. Previous reports indicate that SCCE frequently shows systemic spread and has a poor prognosis. Beta-catenin has been shown to be a key downstream effector of the Wnt signaling pathway, which regulates cell growth and survival. Decreased membranous expression of beta-catenin in cancers correlates with poor prognosis and is associated with dissemination of tumor cells and the formation of metastases. Recently, some different investigators demonstrated aberrant beta-catenin accumulation in neuroendocrine tumors arising in different organs, suggesting a role for the Wnt/beta-catenin signaling pathway during neuroendocrine tumorigenesis. Here, we report a new case of SCCE associated with peritoneal spreading and aggressive course; the patient died one month after surgery. This study also aimed at assessing the involvement of the Wnt signaling pathway in this rare neuroendocrine tumor. Interestingly, both intense nuclear beta-catenin accumulation and cyclin D1 immunoreactivity were restricted to carcinoma cells invading lymphatic vessels. However, mutation analysis failed to demonstrate any mutation in exon 3 of the beta-catenin gene or exon 15 of the APC gene in the present case. Although the mechanism of nuclear accumulation of beta-catenin is still unknown, the heterotopic nuclear localization of beta-catenin may play a role in the tumor invasion process and, subsequently, may be associated with the aggressive behavior of SCCE. PMID- 14533941 TI - Large cell neuroendocrine carcinoma of the urinary bladder with lymphoepithelioma like features. AB - The group of undifferentiated carcinomas of the urinary bladder encompasses small cell undifferentiated carcinoma, giant cell carcinoma, lymphoepithelioma-like carcinoma (LELC), and large cell neuroendocrine carcinoma (LCNEC). These tumors are either pure or can be associated with other components, such as transitional cell carcinoma, squamous cell carcinoma, and adenocarcinoma. We report a case of LCNEC of the urinary bladder in a 54-year-old woman. Histologically, the tumor showed features of LELC; immunohistochemically, the tumor cells reacted to chromogranin A, NSE, and synaptophysin. In addition to these neuroendocrine markers, tumor cells were positive for cytokeratin CAM 5.2 and AE1/AE3, and there was focal positivity for vimentin. In situ hybridization for the detection of Epstein-Barr virus was negative. Despite radical cystourethrectomy and six courses of chemotherapy, the patient developed metastases invading the left inguinal lymph nodes 11 months postoperatively. Currently, 16 months postoperatively, the patient has developed metastases spreading into the lymph nodes of the right ischiorectal fossa; therefore, she is receiving a new cyclus of chemotherapy. There are only three previously reported cases of LCNEC of the urinary bladder, and the significance of neuroendocrine differentiation in non small cell carcinomas at this location remains to be established. However, LELC appears to be a separate clinicopathological entity with sensitivity to chemotherapy and a relatively favorable prognosis. The differentiation between LELC and LCNEC with prominent inflammatory reaction could be of therapeutic relevance. However, in our case, this was possible using immunohistochemistry only. PMID- 14533942 TI - Acute adenoviral infection of a graft by serotype 35 following renal transplantation. AB - Adenoviral infections of immunocompetent patients usually present as self limiting pharyngitis, gastroenteritis, urocystitis, or conjunctivitis. In immunosuppressed patients, development of the illness can be severe, even life threatening or fatal, and therapeutical intervention is difficult. Previous case reports of adenoviral infections after kidney transplantation have described a symptomatology of hemorrhagic cystitis, fever, renal dysfunction, and rarely fatal systemic dissemination. Here we report on a 46-year-old female renal transplant recipient suffering from adenoviral serotype 35 nephritis of the donor organ 29 days after transplantation. In this case, the main symptoms of the adenoviral infection were high fever and progressive renal failure of the transplanted organ. At the peak of the clinical symptoms, owing to histological and immunohistochemical evaluations of a kidney biopsy, we were able to establish the diagnosis in time so that adequate therapy could be employed. Immunosuppression was reduced and modified, and a self-limiting course of the infection was observed, followed by significant improvement of graft function. Subsequent to histological diagnosis, adenoviral particles were isolated from urine and identified as adenovirus serotype 35. Adenoviral nephritis of the transplanted organ should be considered in the differential diagnosis of persistent anuria after kidney transplantation. Our case highlights the importance of applying all possible diagnostic techniques, including histological evaluation of renal biopsies. PMID- 14533943 TI - SSX antigens as tumor vaccine targets in human sarcoma. AB - The efficacy of current standard therapies for the treatment of sarcoma remains limited. With the aim of identifying target antigens relevant to the development of vaccine-based immunotherapy of sarcoma, we have addressed the relevance of tumor-specific antigens encoded by genes belonging to the SSX family as vaccine targets in sarcoma tumors. Expression of SSX-1 to -5 was analyzed in a collection of sarcoma tumors of diverse histological subtypes and in sarcoma cell lines. We found expression of at least one SSX-encoded antigen in 42% of sarcoma tumors, including 5 of 7 different histological subtypes, and in 50% of sarcoma cell lines. SSX-1 was the most frequently expressed family member, followed by SSX-4, 2 and -5. Expression of SSX-3 was detected in only one sample. Importantly, most SSX positive samples co-expressed more than one family member. In addition, assessment of CD8+ T cell recognition of HLA-A2+ SSX-2+ sarcoma cells showed that the latter were efficiently recognized and lysed by SSX-2-specific CTLs. The results of this study indicate that SSX antigens are relevant targets for the development of vaccine-based immunotherapy of sarcoma and encourage the start of vaccination trials using SSX-derived immunogens in sarcoma patients. PMID- 14533945 TI - Clinically significant drug interactions with cholinesterase inhibitors: a guide for neurologists. AB - Cholinesterase inhibitors are the only pharmacological class indicated for the treatment of mild to moderate Alzheimer's disease. These drugs are also being used off label to treat severe cases of Alzheimer's disease or vascular dementia and other disorders. The widespread use of cholinesterase inhibitors raises the possibility of their use in combination regimens, with the subsequent risk of deleterious drug-drug interactions in high-risk populations. The purpose of this review is to present the possible sources of pharmacokinetic or pharmacodynamic drug-drug interactions involving cholinesterase inhibitors. The four cholinesterase inhibitors (tacrine, donepezil, rivastigmine and galantamine) that are currently available have different pharmacological properties that expose patients to the risk of several types of drug interactions of nonequivalent clinical relevance. The principal proven clinically relevant drug interactions involve tacrine and drugs metabolised by the cytochrome P450 (CYP) 1A2 enzyme, as well as tacrine or donepezil and antipsychotics (which results in the appearance of parkinsonian symptoms). The bioavailability of galantamine is increased by coadministration with paroxetine, ketoconazole and erythromycin. It is of interest to note that because rivastigmine is metabolised by esterases rather than CYP enzymes, unlike the other cholinesterase inhibitors, it is unlikely to be involved in pharmacokinetic drug-drug interactions. Care must be taken to reduce the risk of inducing central (excitation, agitation) or peripheral (e.g. bradycardia, loss of consciousness, digestive disorders) hypercholinergic effects via drug interactions with cholinesterase inhibitors. A review of the literature does not reveal any alarming data but does highlight the need for prudent prescription, particularly when cholinesterase inhibitors are given in combination with psychotropics or antiarrhythmics. Possible interactions involving other often coprescribed antidementia agents (e.g. memantine, antioxidants, cognitive enhancers) remain an open area requiring particularly prudent use. PMID- 14533944 TI - Diagnosis and treatment of dysthymia in children and adolescents. AB - Dysthymic disorder is a chronic depressive condition occurring in 0.6-4.6% of children and 1.6-8.0% of adolescents. Although symptoms are less severe than those observed in major depression, childhood-onset dysthymic disorder is characterised by a persistent and long-term depressed or irritable mood (mean episode duration 3-4 years), a worse outcome than major depression and, frequently, comorbid disorders (in around 50% of patients). Long-lasting depressive symptoms seem responsible for long-term disabling consequences on social skill learning, psychosocial functioning and consequent professional life, probably contributing to a higher risk of relapse or development of major depression. Consistently, the first episode of major depression occurs 2-3 years after the onset of dysthymic disorder, suggesting that the latter is one of the gateways to recurrent mood disorders. The primary aims of treatment for dysthymic disorder should be to resolve depressive symptoms, reduce the risk of developing other mood disorders over time and strengthen psychosocial functioning, especially in children and adolescents, in order to prevent the potentially serious sequelae of this disorder. As children with dysthymia often have multiple problems, interventions should involve multiple levels and measures: individual psychotherapy, family therapy/education and pharmacological treatment. Psychotherapeutic techniques, such as cognitive-behaviour therapy and interpersonal therapy, have been found to be efficacious interventions in treating children and adolescents with mild to moderate depression in studies including patients with either dysthmia or double depression. SSRIs are the first line drug treatment for children and adolescents because of their safety, adverse effect profile and ease of use (the safety of paroxetine is currently under investigation). Several nonblind studies have shown the efficacy and good tolerability of SSRIs in children and adolescents with dysthymic disorder, but further research is needed to confirm their efficacy and that of newer antidepressants in the treatment of this disorder. Regardless of whether psychotherapeutic or medical treatments are planned, according to clinical experience, psychoeducational interventions and psychosocial support should be provided to parents and other caregivers during the acute treatment phase to help manage the child's irritable mood and foster a therapeutic alliance and better compliance with treatment. Unfortunately, no studies have focused on continuation treatment of paediatric dysthymic disorder. Given the chronicity, recurrence, psychosocial consequences and peculiar response pattern to treatment of dysthymic disorder, establishing effective 'acute' and 'continuation' interventions in this group of patients should be a priority in mental health management. PMID- 14533946 TI - Pramipexole in routine clinical practice: a prospective observational trial in Parkinson's disease. AB - OBJECTIVE: The mixed dopamine D2/D3 receptor agonist pramipexole is effective as monotherapy in early Parkinson's disease and as adjunctive therapy in advanced disease. Clinical trials suggest that the benefits of pramipexole may extend beyond the relief of motor symptoms (akinesia, rigidity and tremor at rest) to amelioration of depressive symptoms in Parkinson's disease. The aim of this study was to confirm the beneficial effects of pramipexole on the core symptoms of Parkinson's disease (with a focus on tremor), as well as to assess its antidepressant activity, during routine clinical practice. The study also aimed to demonstrate the practicability of the Snaith-Hamilton Pleasure Scale (SHAPS D), the Tremor Impact Scale (TIS) and the Short Parkinson's Evaluation Scale (SPES) under conditions of routine clinical practice. STUDY DESIGN: This was a prospective observational study. PATIENTS: Data for 657 outpatients with Parkinson's disease were collected from German hospitals and specialist practices. The majority of patients were in Hoehn & Yahr stage II or III and were receiving levodopa. METHODS: Pramipexole (Sifrol) was initiated at a dosage of 0.375 mg/day (using a three-times-daily schedule) and titrated upwards, as required, at weekly intervals over a 4-week period to a maximum dosage of 4.5 mg/day (three times daily). Clinical evaluation was performed at baseline, at the end of the titration phase and at the end of maintenance therapy. Patients were assessed via the German questionnaire versions of the physician-assessed SPES, the self-evaluated TIS and the SHAPS-D. Changes in scale scores were evaluated nonparametrically, using the Wilcoxon-matched pairs test. Cronbach's alpha was used as a measure for item consistency. RESULTS: Pramipexole significantly improved SPES subscores for motor symptoms, complications of therapy, psychological status and activities of daily living. Pramipexole also reduced the detrimental effect of tremor on activities of daily living and social interactions, as assessed by patients via the TIS. As indicated by the results of the SHAPS-D questionnaire, pramipexole significantly reduced anhedonia in patients who had associated depression. Internal consistency of SPES subscales was found to be unaltered between the initial evaluation and follow-up. Likewise, internal consistency for TIS and SHAPS-D was demonstrated. Pramipexole was well tolerated and accepted by the vast majority of physicians and patients. CONCLUSION: In addition to ameliorating the core symptoms of akinesia and rigidity in Parkinson's disease, pramipexole improves tremor and depressive symptoms in routine clinical practice. The SPES, TIS and SHAPS-D were found to be useful instruments with validity in this study. PMID- 14533948 TI - Gabapentin: a viewpoint by Brett R. Stacey. PMID- 14533947 TI - Gabapentin: in postherpetic neuralgia. AB - Gabapentin is a structural analogue of the neurotransmitter gamma-aminobutyric acid (GABA) approved for use in adults with postherpetic neuralgia. Gabapentin does not bind to GABA(A) or GABA(B) receptors. Its mechanism of action in humans is unclear, but may involve binding to alpha2delta calcium channel subunits in animal models. Reductions in the mean daily pain score from baseline to week 7 or 8 of treatment (primary endpoint) were significantly greater with gabapentin 1800 3600 mg/day than placebo therapy in two well designed trials in patients with postherpetic neuralgia. The proportion of responders (patients showing a > or =50% reduction in mean daily pain score at endpoint versus baseline) was significantly greater with gabapentin than placebo. Daily sleep rating scores, the Short Form McGill Pain Questionnaire (total pain scores), Patient and Clinician Global Impression of Change and measures on the Short Form-36 Health Survey (including physical functioning, role-physical, bodily pain, vitality or mental health) improved to a significantly greater extent with gabapentin than placebo. Adverse events associated with gabapentin in patients with postherpetic neuralgia were usually mild to moderate in intensity, with dizziness, somnolence and peripheral oedema being commonly reported. PMID- 14533949 TI - Gabapentin: a viewpoint by Ian Gilron. PMID- 14533950 TI - Topiramate: as monotherapy in newly diagnosed epilepsy. AB - Topiramate is an antiepileptic drug that has a broad spectrum of antiseizure effects, which appear to be the result of several neurostabilising pharmacological mechanisms. These include blockade of ion channels, potentiation of GABA neuroinhibition and glutamate receptor antagonism at non-NMDA receptors, as well as mild inhibition of carbonic anhydrase. Topiramate monotherapy dose dependently reduced the number of patients who met seizure related exit criteria in children (aged > or =6 years) and adults with epilepsy. This effect was also observed in patients who had previously experienced partial onset seizures and for those who had experienced generalised tonic clonic seizures. Six-month and 1 year seizure-free rates were dose-dependently reduced. In epilepsy, topiramate monotherapy 100 or 200 mg/day was as effective as carbamazepine 600 mg/day or valproate 1250 mg/day as measured by time to study exit for any reason, time to first seizure and percentage of patients seizure-free in the final 6 months of treatment (mean treatment duration 244 days). Adverse events associated with topiramate monotherapy that were dosage related included paraesthesia, weight loss and diarrhoea. Renal calculi were also reported in both fully published trials. PMID- 14533951 TI - Topiramate: a viewpoint by Nathan Watemberg, MD. PMID- 14533952 TI - Topiramate: a viewpoint by Edward Faught. PMID- 14533954 TI - Endoprosthetic treatment of a mycotic superficial femoral artery aneurysm. AB - PURPOSE: To describe the successful stent-graft exclusion of a mycotic aneurysm of the superficial femoral artery. CASE REPORT: A 78-year-old man presented with Salmonella arteritis and the formation of a mycotic false aneurysm of the superficial femoral artery. Antibiotics were administered; the aneurysm was excluded using 2 Hemobahn stent-grafts, and the surrounding hematoma was drained. One year postoperatively, there are no clinical or biochemical signs of infection. Ultrasound examination does not show any fluid around the patent stent graft. CONCLUSIONS: Stent-graft placement might be an alternative to traditional surgery in selected cases of mycotic aneurysm. PMID- 14533953 TI - Delayed diagnosis of leiomyosarcoma of the common femoral artery after endovascular repair. AB - PURPOSE: To describe a unique case of misdiagnosed leiomyosarcoma of the common femoral artery presenting with signs and symptoms of high-grade stenosis, which was treated with stent placement. CASE REPORT: A 31-year-old woman with a history of diabetes and hyperlipidemia had recurrent claudication and showed significant in-stent restenosis of the common femoral artery in a postoperative angiogram at 5 months. The patient's clinical stage remained unchanged after repeat percutaneous intervention, and leiomyosarcoma was diagnosed from surgical specimens. The patient underwent resection and repair of the involved artery. She has survived 30 months with no further evidence of local recurrence or systemic metastatic disease. CONCLUSIONS: Endovascular repair may cause a long delay in the diagnosis of an arterial leiomyosarcoma mimicking peripheral occlusive disease. PMID- 14533955 TI - Transseptal stent treatment of anastomotic stricture after repair of partial anomalous pulmonary venous return. AB - PURPOSE: To report the endovascular treatment of a stenosis of the pulmonary venous anastomosis following surgical treatment for partial anomalous pulmonary venous return. CASE REPORT: A 60-year-old man presented with recurrent pleural effusions after correction of a partial anomalous left pulmonary venous return. Magnetic resonance imaging demonstrated focal stenosis at the anastomosis of the anomalous pulmonary vein to the left atrial appendage. Using a transseptal approach, the pulmonary vein stenosis was accessed and successfully stented. The patient's symptoms improved, and follow-up imaging demonstrated a patent stent without residual pressure gradient. CONCLUSIONS: Endovascular repair of a stenosis at the anastomosis of an anomalous pulmonary vein is possible and should be considered as a treatment option for this lesion. PMID- 14533956 TI - Hyperperfusion syndrome of the deltoid muscle after subclavian artery angioplasty and stenting. AB - PURPOSE: To report a case of hyperperfusion syndrome of the deltoid muscle after percutaneous transluminal angioplasty of a symptomatic high-grade subclavian artery stenosis. CASE REPORT: Immediately after balloon dilation of a left-sided subclavian artery stenosis, a 53-year-old man developed severe ipsilateral shoulder pain and swelling. Computed tomographic angiography revealed no extravasation or hematoma. Sonography showed massive edema and increased anteroposterior diameter of the left deltoid muscle (3.5 cm compared to 2.0 cm on the right). Hyperperfusion syndrome was suspected, and decompression by anterolateral fasciotomy was performed. Subsequently, both pain and swelling decreased. At day 3, the skin incision, which was temporarily covered with a synthetic skin substitute, was sutured; the wound healed uneventfully. Two weeks after surgery, both muscle strength and shoulder movements showed no restrictions. CONCLUSIONS: Hyperperfusion syndrome after endovascular treatment of subclavian artery stenosis should be considered in the differential diagnosis of atypical muscle pain in the upper extremity. It may present as a compartment syndrome requiring surgical decompression. PMID- 14533957 TI - Transbrachial stenting of a critical ostial arteria lusoria stenosis. AB - PURPOSE: To report the endovascular treatment of a critical ostial stenosis in an aberrant right subclavian artery (ARSA), a rare embryologic anomaly of the aortic arch. CASE REPORT: A 76-year-old woman presented with acute ischemia of her right forearm. Her medical history was notable for hyperlipidemia but otherwise negative for risk factors of atherosclerosis. Color-coded duplex ultrasonography revealed occlusion of the distal brachial and cubital artery, poststenotic flow in the distal subclavian and axillary artery, and reversal of flow in the right vertebral artery. The patient underwent contrast-enhanced multidetector computed tomography of the thorax, which revealed the presence of an ARSA with a calcified, high-grade ostial stenosis. The lesion was successfully stented via a brachial access. CONCLUSIONS: Endovascular treatment of an ostial stenosis in an aberrant right subclavian artery is feasible by means of transbrachial approach. PMID- 14533958 TI - False aneurysm after balloon dilation of a calcified innominate artery stenosis. AB - PURPOSE: To describe the occurrence of a false aneurysm after percutaneous transluminal angioplasty of a severely calcified, high-grade stenosis of the brachiocephalic trunk. CASE REPORT: A 68-year-old patient was admitted with cerebrovascular insufficiency, right upper extremity ischemia, and recurrent amaurosis fugax. Magnetic resonance arteriography (MRA) and duplex ultrasound revealed a severely calcified, high-grade stenosis of the innominate artery. Balloon dilation of this lesion was performed with minimal morphological success. After dilation, the patient suffered a reversible ischemic neurological deficit, from which he recovered completely. The patient's symptoms improved, but after 3 weeks he was re-admitted with palsy of the recurrent laryngeal nerve. An MRA showed a false aneurysm at the dilation site. Open surgery was performed, and the patient received a bifurcated aorto-subclavian-carotid bypass graft. He was discharged free of his preoperative symptoms. CONCLUSIONS: This case illustrates the potential hazards of angioplasty of severely calcified stenoses of the supra aortic vessels. PMID- 14533959 TI - Biocompatibility and biostability of metallic endovascular implants: state of the art and perspectives. AB - More than a million metallic endovascular devices are implanted each year, but the quest for the perfect material continues. The importance of interfacial properties in the overall biocompatibility of metals and alloys has been recognized for a long time. In particular, these properties modulate the hemocompatibility of devices in contact with blood and, in turn, strongly influence implantation outcomes. In this article, the relative properties of metallic materials commonly used in endovascular applications are reviewed. Particular emphasis is given to the corrosion behavior of metallic endovascular materials and the specific surface treatments used in the production processes. Issues relative to corrosion assays will also be reviewed in terms of their relevance to in vivo applications. The potential adverse effects of degradation products with respect to endovascular applications will be described. Finally, this review addresses future perspectives of metallic devices in endovascular procedures in view of the recent promises of antiproliferative strategies that are likely to profoundly modify current procedures. PMID- 14533960 TI - Calcification and proteoglycan accumulation following balloon angioplasty and stenting. AB - PURPOSE: To examine in a porcine model if a correlation exists between calcification and the hyperplastic response of arteries to balloon angioplasty and stenting. METHODS: Eleven Sinclair miniature swine on an atherogenic diet underwent balloon angioplasty or endovascular stenting in nondiseased external iliac arteries using standard procedures. Ninety days postoperatively, the animals were euthanized, and histological sections of the dilated, stented, and control arteries were examined for evidence of calcification and increased proteoglycan accumulation. RESULTS: An increase in proteoglycan accumulation and the loss of integrity of the internal elastic lamina were observed in both the dilated and stented arteries. Diffuse calcification was seen in the media/neomedia of both groups, and large calcium salt deposits were observed in the dilated arteries at the site of the internal elastic lamina and near the stent struts in stented arteries. CONCLUSIONS: Internal elastic lamina loss of integrity appears to be related to an increase in proteoglycans through the formation of a neointima, and the arterial hyperplastic response to balloon angioplasty and stenting seems to lead to calcification. PMID- 14533961 TI - A rapid-exchange monorail stent system for salvage of failing femoropopliteal bypass grafts. AB - PURPOSE: To analyze the safety and effectiveness of a new monorail stent system for the treatment of failing femoropopliteal bypass grafts. TECHNIQUE: Acute distal occlusions or stenoses of femoropopliteal bypass grafts were treated with balloon-expandable stents (13 or 18-mm diameter) pre-mounted on a monorail balloon catheter system. The delivery system was assessed subjectively for (1). compatibility with the sheath, (2). lesion crossing potential, (3). radiopacity, (4). flexibility of the catheter, (5). adequacy of stent expansion, and (5). balloon refolding. In 8 failing bypass grafts with distal lesions, the delivery system successfully deployed the stent at the desired location. Sheath compatibility, catheter flexibility, lesion crossing, and stent expansion were rated "excellent" by all examiners for the 18-mm device. Radiopacity of the mounted stent was graded "good" before and during positioning, but only "sufficient" following expansion. For this type of lesion, the investigators rated the overall performance of the device superior to conventional "over-the wire" systems. CONCLUSIONS: The monorail balloon-expandable stent delivery system provides rapid introduction of the device over the guidewire, and its low profile facilitates the use of small sheaths to minimize access-site complications. PMID- 14533962 TI - Endovascular treatment of malignant superior vena cava syndrome: is bilateral wallstent placement superior to unilateral placement? AB - PURPOSE: To report our experience with unilateral versus bilateral stent placement in the treatment of malignant superior vena cava syndrome (SVCS). METHODS: The records and films of 84 consecutive patients (69 men; mean age 64+/ 10 years, range 39-79) referred for stent placement in malignant SVCS were reviewed for venous compromise, technical and clinical success, complications, and reocclusions. Wallstents were placed covering the SVC and both (bilateral technique) brachiocephalic veins (BCV) preferentially; unilateral stenting of only one BCV in addition to the SVC was performed based on operator preference or inability to access both sides. Technical success was defined as the ability to stent the SVC and at least one BCV; clinical success was the elimination of SVCS symptoms. RESULTS: Technical success was achieved in 83 (99%) patients, using the unilateral technique in 22 and bilateral stenting in 61 patients. The groups did not differ with regard to age, sex, underlying diseases, or location and extent of venous compromise. Immediate clinical success was achieved in 20 (91%) of 22 patients in the unilateral group and 55 (90%) of 61 patients in the bilateral group. Two patients suffered late occlusion in the unilateral group, while in the bilateral group, 8 patients had early occlusion and 9 had late occlusion. Thus, the total occlusion rate was significantly (p<0.05) lower in the unilateral group. There was 1 other complication (pericardial tamponade) in the bilateral group, for a 28% total complication rate, which was significantly higher (p=0.039) than the 9% in the unilateral group. The 1, 3, 6, and 12-month primary stent patency rates were 90%, 81%, 76%, and 69%, respectively. Patency tended to last longer in the unilateral group, but the difference was not significant (p=0.11). CONCLUSIONS: Although bilateral Wallstent placement achieved equal technical and clinical success, it tended to confer shorter-lived patency and caused more complications. PMID- 14533963 TI - The "Y" stent: a technique using nitinol stents to treat bifurcations. AB - PURPOSE: To investigate a new method of stent deployment using commercially available self-expanding stents to treat bifurcation lesions, providing complete lesion coverage without obstructing branches. METHODS: After preliminary in vitro and clinical testing with biliary obstructions, 11 vascular obstructive lesions in 10 patients (7 men; mean age 63 years, range 32-81) were treated by percutaneous Y-shaped stenting. Five patients had iliac arterial stenoses involving the iliac bifurcation; the other 5 patients had 6 central venous obstructions involving branch points. Bifurcated stents were constructed in situ by deploying a nitinol stent extending into one branch and selective balloon fracturing of struts to create a fenestration. A second stent was deployed through the hole, followed by symmetrical fracturing of struts in the second stent to form a Y-shaped configuration. RESULTS: All deployments were technically successful and resulted in complete coverage of the bifurcation lesions. Protrusion of fractured stent material into flow lumens was minor as determined by angiography, intravascular ultrasound, and passage of balloons, but it required the stents to be oversized. The lengths and luminal diameters were tailored for each arm of the Y-shaped stents. CONCLUSIONS: In situ creation of a Y-shaped stent using 2 self-expanding nitinol stents is a feasible option with potential advantages in the treatment of obstructive lesions involving a bifurcation. PMID- 14533964 TI - Clinical experience with the OptiMed sinus stent in the peripheral arteries. AB - PURPOSE: To retrospectively evaluate the performance of a new self-expanding nitinol stent in the treatment of lower limb arterial occlusive lesions. METHODS: The Sinus stent is a flexible, laser-cut device with good radiopacity and radial strength, allowing precise placement via a 7-F introducer, even in tortuous arteries. Over a 3.5-year period, 315 patients (254 men; mean age 64.9+/-10.8 years, range 39-93) had 403 lower limb lesions treated: 172 iliac (132 stenoses, 40 occlusions), 204 femoral (131 stenoses, 73 occlusions), and 27 popliteal (19 stenoses, 8 occlusions). The majority of patients (272, 86%) were in stage IIb of Fontaine's classification, 31 (10%) in stage III, and 12 (4%) in stage IV. Mean lesion lengths were iliac: 51.9+/-32.1 mm, femoral: 66.3+/-51.5 mm, and popliteal: 44.3+/-27.2 mm. Indications for stenting were 222 postdilation residual stenoses, 125 dissections, and 35 restenoses; 21 lesions were directly stented. RESULTS: Immediate technical success was 100%. Clinical success was 98%. The ankle-brachial index increased from 0.62+/-0.12 to 0.91+/-0.14. In 64 lesions, there were 3 early thromboses. Mean follow-up was 16.1+/-9.7 months (range to 40). There were 79 restenotic episodes: 14 iliac, 56 femoral, and 9 popliteal. Primary and secondary patencies for all lesions at 3 years were 72.5%+/-6.6% and 81.8%+/-5.9%, respectively. Stenoses demonstrated better patency than occlusions (primary: 78.0%+/-7.3% versus 60.2%+/-7.1%, p<0.001; secondary: 86.8%+/-6.1% versus 71.3%+/-6.7%, p<0.001). Men had better secondary patency at 3 years (83.7% versus 73.4%, p<0.05). For femoral lesions >8 cm, the results were acceptable for stenoses (66.7% primary and secondary patency at 3 years), but for lengthy occlusions, the restenosis rate was high (50.0% for occlusions versus 25.0% for stenoses, p<0.05). CONCLUSIONS: Angioplasty with the Sinus stent seems safe and effective in the treatment of peripheral arterial occlusive diseases, giving excellent results at the iliac and common femoral artery levels. Treatment results are better for stenoses than for occlusions. An occlusion length 4 showers of microemboli at postdilation (odds ratio [OR] 3.2, 95% CI 1.3 to 7.8, p=0.03), particulate macroemboli (OR 9.1, 95% CI 5.1 to 16.1, p<0.001), massive air embolism from ruptured balloons (OR 11.3, 95% CI 7.6 to 16.6, p<0.001), and angioplasty-induced asystole with significant hypotension plus MCA blood flow reduction (OR 3.3, 95% CI 1.4 to 8.3, p=0.03). Of the patient characteristics, male gender (OR 10.5, 95% CI 1.4 to 75.8, p=0.02) and preoperative cerebral ischemia (OR 3.3, 95% CI 1.6 to 6.6, p=0.003) were also related to outcome. CONCLUSIONS: In CAS, TCD monitoring provides insight into the pathogenesis of procedure-related cerebral events. Microemboli during poststent dilation, particulate macroembolism, massive air embolism, and angioplasty-induced asystole are associated with adverse outcome, as are male gender and prior cerebral ischemia. PMID- 14533975 TI - Belgian experience with FilterWire EX in the prevention of embolic events during carotid stenting. AB - PURPOSE: To evaluate the performance of the FilterWire EX for the capture and removal of debris and prevention of embolic events during endovascular carotid interventions. METHODS: In the period from October 2001 to May 2002, 100 patients (60 men; mean age 72 years, range 48-90), 69 symptomatic with >or=50% internal carotid artery (ICA) stenosis and 31 asymptomatic with >or=70% stenosis were scheduled for carotid stenting with cerebral protection provided by the FilterWire EX system. Satisfactory performance of the filter was defined as effectiveness in preventing intraprocedural embolic events shown by visible debris in the filter and unchanged neurological status assessed by an independent neurologist. RESULTS: The placement and retrieval of the FilterWire EX was technically successful in 93 of 100 patients. Five delivery failures were due to severe ICA angulation; one stenosis was underestimated on preprocedural imaging, and one case was aborted following a major stroke during sheath manipulation before filter introduction. In 56.9% of the protected cases, the filter contained debris. One neurological event occurred 6 hours after the procedure: a transient ischemic attack from which the patient recovered fully after 2 hours. No other procedure-related events were recorded within 30 days of the procedure. The procedural success among the 93 patients stented under FilterWire EX protection was 98% (92% by intention to treat). CONCLUSION: The FilterWire EX appears to be an effective tool in the prevention of embolic events and in preserving neurological status during endovascular treatment of carotid artery stenosis. PMID- 14533976 TI - Carotid stenting versus carotid surgery: a prospective cohort study. AB - PURPOSE: To compare the results of carotid stenting and carotid surgery in a consecutive group of 455 patients (482 lesions). METHODS: Between January 1995 and July 2002, 107 patients (114 lesions) were treated with carotid stenting and 348 patients (368 lesions) with carotid surgery. A cerebral protection device was routinely used in the last 46 stent cases. All patients were followed with duplex examination at 1 and 6 months postoperatively and yearly thereafter. RESULTS: The stent and surgery groups were similar in terms of mean age (70.5 and 71.1 years, respectively), sex distribution (men 72% versus 75%), and symptoms (transient ischemic attack [TIA] or minor stroke 32.7% versus 42.2%). Median follow-up was 15 months in stent patients and 14 months in the surgical group. At 1 month postoperatively, there were 7 minor strokes (5 temporary lasting <21 days and 2 persistent) and 1 major stroke in the stent group versus 1 persistent minor stroke in the surgical patients. The overall neurological event rate (including TIA) was 10.5% versus 1.9% (p=0.0002) in the surgical patients; cardiac morbidity was 3.5% versus 1.6% (p=NS), and the death rate was 0% versus 0.8% (p=NS). The combined permanent stroke/death rate was 2.6% in stent patients and 1.1% in surgery patients (p=NS). During follow-up, the cumulative all stroke rate was 8.8% versus 1.9% (p=0.001), but the 3-year cumulative survival rate free from ipsilateral major neurological events was 95.2% in stent patients and 96.9% in the surgery cohort (p=NS). There was a 7.5% rate of restenosis in stented arteries versus 1.4% in surgery patients (p=0.001). CONCLUSIONS: This series showed encouraging and comparable major stroke and deaths rate of carotid stenting and carotid surgery. However, there was more restenosis in the stented group on midterm follow-up. PMID- 14533978 TI - A feeling in the waters: diagnosis of heart failure using urinary natriuretic peptides. AB - Plasma levels of brain natriuretic peptide (BNP) and N-terminal pro-BNP (N-BNP) are highly sensitive markers of ventricular dysfunction and/or hypertrophy and, in established disease, offer prognostic value and may be useful for guidance of therapy. Ng and co-workers report in this issue of Clinical Science that urinary levels of N-BNP may be as useful as plasma levels for the discrimination of patients with and without heart failure. This raises the potential for a relatively simple urine test that could be used for the diagnosis of heart failure. Roles in prognostication and the guidance of therapy may also be possible but, perhaps of most significance, measurement of urinary N-BNP may be applied to screening of patients at high risk of heart failure. The main limitations of the study were that the sample of heart failure patients comprised only 34 individuals with New York Heart Association functional Class IV and that the observed correlation between levels of urinary N-BNP and plasma creatinine seemed counter-intuitive. The latter issue needs clarification, as renal impairment is a frequent co-morbidity among patients with heart failure and will potentially confound any observed association between ventricular dysfunction and urinary N-BNP levels. Another caveat is that it is unclear if testing for urinary N-BNP can be cheaply and conveniently administered on a large scale. Nevertheless, this first demonstration of elevated N-BNP in the urine of patients with heart failure raises a number of exciting possibilities with regard to the management of patients with established or possible heart failure. Further investigation is required and eagerly awaited. PMID- 14533979 TI - Catalytic- and ecto-domains of membrane type 1-matrix metalloproteinase have similar inhibition profiles but distinct endopeptidase activities. AB - Membrane type 1-matrix metalloproteinase (MT1-MMP/MMP-14) is a major collagenolytic enzyme that plays a vital role in development and morphogenesis. To elucidate further the structure-function relationship between the human MT1 MMP active site and the influence of the haemopexin domain on catalysis, substrate specificity and inhibition kinetics of the cdMT1-MMP (catalytic domain of MT1-MMP) and the ecto domain DeltaTM-MT1-MMP (transmembrane-domain-deleted MT1 MMP) were compared. For substrate 1 [Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH(2), where Mca stands for (7-methoxycoumarin-4-yl)acetyl- and Dpa for N -3-(2,4 dinitrophenyl)-L-2,3-diaminopropionyl], the activation energy E (a) was determined to be 11.2 and 12.2 kcal/mol (1 cal=4.184 J) for cdMT1-MMP and DeltaTM MT1-MMP respectively, which is consistent with k (cat)/ K (M) values of 7.37 and 1.46x10(4) M(-1).s(-1). The k (cat)/ K (M) values for a series of similar single stranded peptide substrates were determined and found to correlate with a slope of 0.17 for the two enzyme forms. A triple-helical peptide substrate was predicted to have a k (cat)/ K (M) of 0.87x10(4) M(-1).s(-1) for DeltaTM-MT1-MMP based on the value for cdMT1-MMP of 5.12x10(4) M(-1).s(-1); however, the actual value was determined to be 2.5-fold higher, i.e. 2.18x10(4) M(-1).s(-1). These results suggest that cdMT1-MMP is catalytically more efficient towards small peptide substrates than DeltaTM-MT1-MMP and the haemopexin domain of MT1-MMP facilitates the hydrolysis of triple-helical substrates. Diastereomeric inhibitor pairs were utilized to probe further binding similarities at the active site. Ratios of K (i) values for the inhibitor pairs were found to correlate between the enzyme forms with a slope of 1.03, suggesting that the haemopexin domain does not significantly modify the enzyme active-site structure. PMID- 14533980 TI - Investigating conformational stability of bovine pancreatic phospholipase A2: a novel concept in evaluating the contribution of the 'native-framework' of disulphides to the global conformational stability of proteins. AB - Bovine pancreatic PLA(2) (phospholipase A(2)) is a 14 kDa protein whose structure is highly cross-linked by seven disulphide bonds. We investigated the structural stability of this enzyme by the method of 'disulphide-scrambling' with denaturants such as urea, GdmCl (guanidine hydrochloride), GdmSCN (guanidine thiocyanate) and at high temperatures in the presence of 2-mercaptoethanol (0.2 mM) as thiol initiator. Reverse-phase HPLC was used to follow denaturation. To denature 50% of the native protein, 1.25 M GdmSCN, approx. 3 M GdmCl and higher than 8 M urea were required. Only 20% of the protein was denatured after 2 h at 60 degrees C, whereas complete denaturation was seen after 2 h at 70 degrees C and within 30 min at 80 degrees C. A distinct enhancement of stability was observed when denaturation was conducted in the presence of 10 mM calcium chloride, which has not been reported previously. CD studies of GdmCl denaturation of bovine PLA(2) showed that 2.5 M GdmCl was required to denature 50% of the protein in the presence of 0.2 mM 2-mercaptoethanol (in agreement with the HPLC analysis), whereas 6.4 M GdmCl was necessary to denature 50% of the protein in the absence of a thiol initiator. Conformational stability (Delta G (water)) was estimated to be 8.7 kcal/mol (1 cal=4.184 J) by 'disulphide-intact' denaturation (where 'native' disulphide framework was unaffected) and 2.5 kcal/mol by 'disulphide-scrambling' denaturation (involved breaking of native disulphides and formation of 'non-native' ones). The difference, Delta(Delta G (water)), of 6.2 kcal/mol was the conformational stability contributed by the 'native-framework' of seven disulphides. Using bovine PLA(2) as an example, we have demonstrated a novel comparative technique, where the conformational stability study of a disulphide-containing protein, with a common denaturant, in both the presence and absence of catalytic amounts of a thiol initiator can be used as a convenient method to estimate selectively and quantitatively the actual contribution of the 'native disulphide bond network' towards the global conformational stability of the protein. PMID- 14533981 TI - Favorable patient acceptance of ambulatory blood pressure monitoring in a primary care setting in the United States: a cross-sectional survey. AB - BACKGROUND: The use of ambulatory blood pressure monitoring (ABPM) in the diagnosis and management of hypertension in primary care settings in the United States is increasing. Insufficient information is available describing patients' experiences and acceptance of this technology in the United States, where medical insurance coverage of the procedure is often limited. The objective of this study was to describe patient satisfaction with ABPM performed in a primary care office in the United States, using modern ABPM technology. METHODS: Cross-sectional survey performed on consecutive patients referred to the ABPM service of the Family Care Center, University of Iowa Hospitals and Clinics, Iowa City, Iowa from January 2001 to July 2003. Measures of patient satisfaction and acceptance with the device, comfort, and overall session were assessed via a 9-question, Likert-scale response survey. RESULTS: Since its inception two and a half years ago, 245 total ABPM sessions have been conducted in 235 unique patients. Of the 235 eligible respondents, 177 returned completed surveys, yielding a 75% response rate. Three-fourths (75%) of patients believed that undergoing the test was worthwhile considering the time and monetary cost involved, while most (90%) reported they thought the information provided by the test would be helpful to their physician in making treatment decisions. Patients reporting that their physician had clearly explained the benefit of undergoing the testing were more likely to report that they thought the results of the test would be more helpful in making treatment decisions. Few patients (20%) found that wearing the monitor was uncomfortable. CONCLUSIONS: When clinically indicated, clinicians should not hesitate to order ABPM testing for fear of subjecting patients to an uncomfortable test, or an uncovered insurance benefit. When ordering ABPM, they should be sure to educate the patient about the potential benefits of undergoing the testing. Most patients believe the test will provide useful information in making treatment decisions, despite probable lack of insurance coverage, and appear willing to experience some discomfort for the overall gain of the results obtained from undergoing the session. PMID- 14533982 TI - An outbreak of conjunctivitis caused by multiresistant Pseudomonas aeruginosa in a Brazilian newborn intensive care unit. AB - We report an outbreak of conjunctivitis due to Pseudomonas aeruginosa involving seven infants admitted in the Neonatal Intensive Care Unit (NICU) of the Uberlandial Federal University Hospital between March and September 2001. Three infants developed systemic complications (01 sepsis and 02 pneumonias). Ten isolates were obtained from conjunctival cultures and all were resistant to ceftazidime and aminoglycosides. Fast identification of the organism and treatment with imipenem were important in containing the outbreak of P. aeruginosa. PMID- 14533983 TI - Frequency of polymorphisms of genes coding for HIV-1 co-receptors CCR5 and CCR2 in a Brazilian population. AB - Entry of human immunodeficiency type 1 virus (HIV-1) into target cells requires both CD(4)and one of the chemokine receptors. Viruses predominantly use one, or occasionally both, of the major co-receptors CCR5 and CXCR4, although other receptors, including CCR2B and CCR3, function as minor co-receptors. A 32 nucleotide deletion (D32) within the b-chemokine receptor 5 gene (CCR5) has been described in subjects who remain uninfected despite extensive exposition to HIV 1. The heterozygous genotype delays disease progression. This allele is common among Caucasians, but has not been found in people of African or Asian ancestry. A more common transition involving a valine to isoleucine switch in transmembrane domain I of CCR2B (64I), with unknown functional consequences, was found to delay disease progression but not to reduce infection risk. As the Brazilian population consists of a mixture of several ethnic groups, we decided to examine the genotype frequency of these polymorphisms in this country. There were 11.5% CCR5 heterozygotes among the HIV-1 infected population and 12.5% among uninfected individuals, similar to data from North America and Western Europe. The prevalence of CCR2-64I homozygotes and heterozygotes was 0.06 and 15.2%, respectively, also similar to what is known for North America and Western Europe. PMID- 14533984 TI - C-reactive protein in the diagnosis of community-acquired pneumonia. AB - Qualitative determination of C-reactive protein (CRP) was evaluated as a diagnostic method for community-acquired pneumonia. Paired serum and pleural fluid samples from child patients were examined with a CRP test, compared to bacterial cultures, counterimmunoelectrophoresis and immunoassay. The CRP test gave excellent parameters of sensitivity, specificity and predictive values for the diagnosis of bacterial pneumonia. PMID- 14533985 TI - Use of Rhu-GM-CSF in pulmonary tuberculosis patients: results of a randomized clinical trial. AB - It has been postulated that deficient or incomplete clinical and/or microbiological response to tuberculosis treatment is associated with cell mediated immunological dysfunction involving monocytes and macrophages. A phase 2 safety trial was conducted by treating patients with either recombinant human granulocyte-macrophage colony-stimulating factor (rhu-GM-CSF) or a placebo, both in combination with anti-tuberculosis chemotherapy. Thirty-one patients with documented pulmonary tuberculosis were treated with rifampin/isoniazid for six months, plus pyrazinamide for the first two months. At the beginning of treatment, rhu-GM-CSF (125mg/M(2)) was randomly assigned to 16 patients and injected subcutaneously twice weekly for four weeks; the other 15 patients received a placebo. The patients were accompanied in the hospital for two weeks, then monthly on an out patient basis, for 12 months. Clinical outcomes were similar in both groups, with no difference in acid-fast bacilli (AFB) clearance in sputum at the end of the fourth week of treatment. Nevertheless, a trend to faster conversion to negative was observed in the rhu-GM-CSF group until the eighth week of treatment (p=0.07), after which all patients converted to AFB negative. Adverse events in the rhu-GM-CSF group were local skin inflammation and an increase in the leukocyte count after each injection, returning to normal 72 hours after rhu-GM-CSF injection. Three patients developed SGOP and SGPT > 2.5 times the normal values. All patients included in the GM-CSF group were culture negative at six months, except one who had primary TB resistance. None of the patients had to discontinue the treatment in either group. We conclude that rhu GM-CSF adjuvant immunotherapy could be safely explored in a phase 3 trial with patients who have active tuberculosis. PMID- 14533986 TI - Viral hepatitis in patients infected with human immunodeficiency virus. AB - From 1992 to 1995 we studied 232 (69% male, 87% Caucasian) anti-human immunodeficiency virus (anti-HIV) positive Brazilian patients, through a questionnaire; HIV had been acquired sexually by 50%, from blood by 32%, sexually and/or from blood by 16.4% and by an unknown route by 1.7%. Intravenous drug use was reported by 29%; it was the most important risk factor for HIV transmission. The alanine aminotransferase quotient (qALT) was >1 for 40% of the patients, 93.6% had anti-hepatitis A virus antibody, 5.3% presented hepatitis B surface antigen, 44% were anti-hepatitis B core antigen positive and 53.8% were anti hepatitis C virus (anti-HCV) positive. The anti-HCV test showed a significant association with qALT>1. Patients for whom the probable HIV transmission route was blood had a 10.8 times greater risk of being anti-HCV positive than patients infected by other routes. Among 30 patients submitted to liver biopsy, 18 presented chronic hepatitis. PMID- 14533987 TI - Seroprevalence of HbsAg, Anti-HBc and anti-HCV in Southern Brazil, 1999-2001. AB - The prevalence of infection by the hepatitis B (HBV) and C (HCV) viruses varies among geographical regions. We evaluated 263,795 blood donor samples collected from 1999-2001 in various cities in the state of Santa Catarina to determine the prevalence of HbsAg, anti-HBc and anti-HCV markers. The markers were analyzed by immunoenzymatic tests, as determined by the Ministry of Health, and the data were obtained from blood banks and from ANVISA (the Brazilian National Agency for Sanitary Vigilance). There was a significant reduction in the mean frequency of HbsAg and anti-HBc during the study period, from 0.98% to 0.64% and from 8.83% to 5.35%, respectively, though they varied considerably among the different regions. There was also a decrease in the mean frequency of anti-HCV, although it was not significant, decreasing from 0.38% to 0.34%. Even with this reduction, the frequency of these markers was still high compared with that found in other countries, indicating high rates of infection by hepatitis B and C viruses. This emphasizes the urgency of vaccination programs against HBV, especially in some regions of Santa Catarina state, in order to reduce the prevalence of this infection and consequently reduce the risk of transmission through sexual relations or from the donation of blood and/or hemocomponents. PMID- 14533988 TI - A case study of disseminated histoplasmosis linked to common variable immunodeficiency. AB - Histoplasma capsulatum, a ground fungus, can infect humans, normally in endemic areas; the resulting disease can be asymptomatic or it can have a benign development, but in rare cases it can develop into a serious clinical condition and can even be fatal. Its most characteristic initial location is in the lungs, resembling tuberculosis, often accompanied by mediastinitis and an exuberant fibrotic response. The spread of this infection can be caused by the concomitance of another illness that alters the immunological balance. Sometimes such an association is not clear. Therefore, disseminated histoplasmosis is defined as a clinical condition where the fungus is present in more than one location. Common variable immunodeficiency is characterized by a generalized failure in the synthesis of antibodies, leading the affected individuals to present recurrent infections, especially those caused by encapsulated bacteria, most often involving the respiratory tract. We studied a serious case of disseminated histoplasmosis, accompanied by common variable immunodeficiency, observed at the Infirmary of the department of Medical Practice of the Federal University at Parana Hospital das Clinicas. PMID- 14533989 TI - Atypical presentation of syphilis in an HTLV-I infected patient. AB - We report the case of a 44 year-old female, who presented a long-lasting, clinically atypical, secondary syphilis ("malignant syphilis") in the right foot, which started six months before medical evaluation. The patient had a serological diagnosis of HTLV-I infection and syphilis two years before the onset of the skin lesions, following a blood donation. As she believed she was allergic to penicillin, she initially received sulfamethoxazole + trimethoprim, without any improvement of the clinical picture. After failure of this first treatment regimen, she was given penicillin, which promoted complete healing of the lesion. We found evidence that infection by HTLV-I is capable of modifying the clinical course of secondary syphilis. PMID- 14533990 TI - First isolation of enterovirus 71 (EV-71) from Northern Brazil. AB - Enterovirus 71 (EV-71) has been associated to cases of neurological disease in many countries including Brazil. This virus has now been reported from three of the five Brazilian regions. Our study relates the findings concerning to the first isolate of this virus in Northern region of Brazil. A 15-month old female patient, from the rural zone of the municipality of Santana do Araguaia in southern Para state was admitted at the hospital with acute, flaccid, asymmetric and ascending motor deficiency, located in the right lower limb. Stools samples from this child were inoculated in RD cells and was isolated an EV-71. We plan to sequence our strain and compare it to other isolates in Brazil. Differences at the molecular level can explain why EV-71 strains circulating in other continents, such as Asia, appear to be more virulent. PMID- 14533991 TI - [The public health cost of chronic obstructive pulmonary disease]. PMID- 14533992 TI - [Delays in the diagnosis of lung cancer]. AB - OBJECTIVE: To study the clinical and demographic factors associated with delays in the diagnosis of lung cancer. PATIENTS AND METHODS: A 2-year prospective study of patients admitted to the respiratory medicine ward with a suspected diagnosis of lung cancer. We studied demographic factors, health care received, place of residence, and delays in carrying out diagnostic procedures. The following diagnostic time periods were defined: consultation (from first symptom to first medical visit), middle period (from first medical visit to hospital admission) and diagnostic (from hospital admission to histological diagnosis and clinical staging). RESULTS: One hundred thirteen patients with a mean age of 65 years (range, 36-90), 103 men and 10 women, were studied. The most frequent symptoms leading to consultation were coughing (10.6%), hemoptysis (19.5%), chest pain (26.5%), and shortness of breath (9.7%). First visits were to a primary care physician for 72%, to the hospital emergency room for 22%, or to a pulmonologist for 6%. Forty-four percent of the patients visited the doctor 2 or 3 times. The mean SD, numbers of days for the different time periods were as follows: consultation, 30.3 60; diagnosis, 18.6 19; middle period 37.9 63. The mean total time from first symptom to diagnosis was 85.7 87 days. The middle period, the time in hospital until diagnosis, and the total time were shorter when patients were referred by the primary care physician to the emergency room or were directly admitted to the hospital (P<.001). Only 25.7% of the staged lung cancers were operable. CONCLUSIONS: Delays in lung cancer diagnosis are long. The attitudes of primary care physicians and their relations with specialized care providers are crucial for reducing delays. PMID- 14533993 TI - [Variations in the use of hospital resources in treating patients with chronic obstructive pulmonary disease]. AB - AIM: To analyze factors associated with the length of stay and pharmaceuticals expense for patients admitted with chronic obstructive pulmonary disease (COPD) and to evaluate whether treatment by different physicians means greater or less use of hospital resources. METHODS: We retrospectively studied a cohort of 1033 COPD patients admitted to the Hospital de Mataro, Catalonia, Spain, during the years 1996 through 1998 in order to analyze factors associated with length of stay and pharmaceuticals use. We used the Minimum Basic Data Set, laboratory databases, and pharmacy single-dose database. We also analyzed the differences among patients treated by different physicians and developed multiple linear regression models to evaluate differences in treatment between one physician and another. RESULTS: The length of stay increased with patient age, the number of times admitted, the presence of atrial fibrillation or respiratory insufficiency, ventilatory alterations, chronicity, a forced expiratory volume in 1 second less than 50% of predicted, and treatment by certain physicians. Medication costs showed a similar pattern. Although the characteristics of patients treated by different physicians were quite homogeneous, the median length of stay varied from 9 to 11 days depending on the physician, while the median cost for medication varied from;43.62 to;54.39 (from $41.07 to $51.21). After removing the effects of several covariables by multiple regression analysis, an effect of physician persisted. CONCLUSIONS: Significant differences in length of hospital stay and consumption of pharmaceuticals are related to attending physician and continue to have an important effect after controlling for the differences in the severity of patient status. PMID- 14533994 TI - [Clinical predictors of sleep apnea-hypopnea syndrome susceptible to treatment with continuous positive airway pressure]. AB - OBJECTIVE: To analyze the predictive value of clinical data for identifying patients suspected of sleep apnea-hypopnea syndrome with an apnea-hypopnea index (AHI)> or = 30. MATERIAL AND METHODS: Patient characteristics, cardiorespiratory medical history, and clinical signs and symptoms were recorded for all patients. Exclusion criteria were daytime respiratory insufficiency or heart failure. All patients underwent polysomnographic testing (AutoSet Portable Plus II, ResMed Corp, Sydney, Australia) for automatic AHI calculation and manual determination of central and obstructive apneas. A logistic regression model was constructed to calculate the likelihood of an individual's presenting an AHI> or = 30 as well as the predictive value of each variable and of the final model. RESULTS: Three hundred twenty-nine patients with a mean +/- SD age of 58 +/- 13.45 years were studied; 76.4% were men. Data for 207 patients were used to construct the logistic regression model: logit (P) = 2.5 blood pressure + 1.5 Epworth test + body mass index + 0.6 repeated observed episodes of apnea 2.1. Logit(P) was loge (1-P)/P and variables were dichotomized with cut points of 11 for the Epworth test and of 30 kg/m2 for body mass index. The diagnostic sensitivity of the model was 80.2% (75%-86%), specificity was 93.4% (89%-95%), positive predictive value was 89.6% (84%-93%) and negative predictive value was 86.9% (81%-90%), such that 89.6% of the patients were correctly classified. The variable with the greatest predictive value was high blood pressure. The model was validated prospectively in the remaining 102 patients. CONCLUSIONS: Prior to diagnostic tests for SAHS, clinical data can be useful for identifying patients suspected to have a AHI> or = 30. PMID- 14533995 TI - [Tuberculosis control in the Central Health Region of Catalonia during the period 1986-2000]. AB - OBJECTIVE: To assess tuberculosis control in the Central Health Region, Catalonia, Spain, from the implementation of the area's Tuberculosis Control Program in 1986 until the year 2000. METHOD: To study the epidemiological profile of tuberculosis and the outcome of the following control measures in sputum smear positive patients: final outcome of treatment and monitoring, and the percentage of patients for whom a contact investigation (CI) was carried out. Tuberculosis control is considered effective if the sum of noncompliant patients, plus the cases in which treatment failed, plus the patients transferred out remains below 10% of the cohort of patients studied, and if a CI has been conducted in at least 80% of this cohort. RESULTS: The number of cases reported was 6326, of which 7% were retreatments. A total of 5865 new cases was detected. Of these, 5652 (96%) were patients born in Spain. The number of cases reported annually peaked in 1990 (474) and then declined continuously until 2000, when it was 54% lower (220). Foreign-born patients numbered 213 (4%), and 95% of them came from countries with a high prevalence of tuberculosis. In the cohort of patients studied between 1997 and 2000, the sum of noncompliant patients, plus cases in which treatment failed, plus the patients transferred out came to 11%. Since 1994, a CI has been carried out for over 80% of patients, and this figure reached 92% in 2000. CONCLUSIONS: We consider that a substantial degree of tuberculosis control has been achieved given the decline in tuberculosis morbidity among people born in Spain, the fact that the percentage of noncompliant patients, treatment failures and transfers was only slightly over 10%, and that a CI has been conducted for over 80% of patients since 1994 (92% in 2000). PMID- 14533996 TI - [Series 4: respiratory muscles in neuromuscular diseases and the chest cavity. Pathophysiological consequences of lung volume reduction surgery in patients with emphysema]. PMID- 14533997 TI - [Consensus conference: home intravenous antibiotic treatment for cystic fibrosis. The Sira Carrasco Foundation. 26 April 2002]. PMID- 14533998 TI - [Sildenafil as a substitute for subcutaneous prostacyclin in pulmonary hypertension]. AB - Subcutaneous prostacyclin (treprostinil) is an effective short-term treatment for pulmonary hypertension. The most frequently described adverse effect-pain in the area of injection-rarely requires that treatment be withdrawn. Sildenafil is a selective fosfodiesterase-5 inhibitor with pulmonary vasodilating effects. We describe the use of sildenafil as a substitute for treprostinil in a patient with pulmonary hypertension associated with lupus erythematosus. Treatment with treprostinil was discontinued due to uncontrollable abdominal pain. PMID- 14533999 TI - [Pulmonary infection due to Nocardia nova]. PMID- 14534000 TI - [Recurrent tuberculous pneumothorax and tuberculous empyema: an association of two rare complications]. PMID- 14534001 TI - [What should surgeons explain to their patients?]. PMID- 14534004 TI - [Value of brain perfusion SPECT in the follow-up of patients surgically treated for chronic adult hydrocephalus syndrome]. AB - OBJECTIVE: The aim of this study was to characterize regional cerebral blood flow in patients with Adult Hydrocephalus Syndrome (AHS) and to evaluate the changes in brain perfusion after surgical derivation treatment. PATIENTS AND METHODS: 20 patients with AHS (age: 72 +/- 14, 12 men) were studied before and six months after surgery. All patients underwent a brain perfusion SPECT (99mTc-HMPAO) prior to surgery and at 6 months post-surgery. Semi-quantitative analysis was done for brain uptake: 0=Normal, 1=Mild, 2=Moderate, 3= Severe, 4=No uptake. The severity of ventricular dilatation was assessed by classifying the intensity and extension of subcortical defects: 0=Normal, 1=Mild, 2=Moderate, 3=Severe. The scores of the pre- and post-surgical studies were compared using the Student-t test. RESULTS: A global reduction of brain uptake was observed (mean score 12.85), mainly in frontal, parietal and temporal lobes, with a significant improvement in post surgical studies (mean score 6, p<0,001). After surgery, 16 (80%) of the 20 patients improved brain uptake. In relationship to subcortical uptake, 5 patients showed mild defects, 9 moderate defects and 6 patients presented severe uptake reduction. In post-surgical studies 15 (75%) patients improved almost one degree in the subcortical score and 65% of the patients showed a normal or mild subcortical uptake reduction. CONCLUSIONS: Brain perfusion SPECT is useful in patients with AHS, detecting brain perfusion defects and evaluating cerebral blood flow improvement after shunt operation. PMID- 14534006 TI - [Improvement of the imaging quality by a non-uniform attenuation correction system in cardiac SPECT]. AB - A non-uniform attenuation correction system has been purchased recently by the Department of Nuclear Medicine of the University Hospital in Salamanca to be used in a dual-detector Picker Axis gammacamera. This system is based on the generation of an attenuation map from each patient using a transmission scan with and without the patient using two Ba-133 sources. At present, this system is only available for a 102 configuration between the detectors so its use is restricted to cardiac single photon emission computed tomography (SPECT). The aim of this work has been to evaluate improvement of the image quality of this attenuation correction system by doing three different tests (evaluation of the recovery coefficient, activity concentration ratio and attenuation residual error). After analyzing all the tests, the results for the non-uniform attenuation correction system have been favorable compared to the conventional correction method employed in the clinical practice. PMID- 14534005 TI - [Recombinant human thyrotropin in the follow-up of patients with differentiated thyroid carcinoma]. AB - PURPOSE: To determine the value of the use of radio-iodine scanning diagnostic and radio-iodine therapy during the follow-up of advanced differentiated thyroid cancer without stopping thyroid hormone suppression therapy. MATERIAL AND METHODS: We performed 7 radio-iodine scans and 4 radioiodine therapies in 5 men and 1 woman, aged 39 to 79 years. Five patients had papillary thyroid cancer and one follicular thyroid cancer. Human recombinant thyrotropin was given intramuscularly at a dose of 0.9 mg/ml once a day for two days. Twenty-four hours after the second dose, serum thyrotropin, thyroglobulin (Tg) and thyroglobulin autoantibodies were measured and each patient was given 2 mCi of 131I in postsurgery scans and 5 mCi of 131I in patients previously treated with 131I. Whole-body scans were obtained within 54 hours of injection and in the cases of diagnostic uncertainty a second scan was performed 72 hours after the injection. RESULTS: Four scans were positive and two were negative. One positive scan patient was not treated due to a serious clinical condition. An effective increase of serum TSH was observed in all 6 patients. Four patients had an effective increase of serum Tg; the other 2 cases had a decrease of serum Tg: One case was undifferentiated and in the other case the mass was smaller after surgery. CONCLUSIONS: rhTSH is an effective means of stimulating TSH levels and allowing scan and/or radioiodine treatment. rhTSH has no significant adverse effects. The use of rhTSH avoids all the disadvantages of hypothyroidism and maintains quality of life. PMID- 14534007 TI - [Follow-up of children and young adults with differentiated thyroid cancer treated with radioiodine]. AB - This is a retrospective study carried out in a group of 30 patients with differentiated thyroid cancer (age at diagnosis equal to or less than twenty years old). The aim of the study is to evaluate outcome after 131I therapy. Patients were classified into three groups on the basis of initial surgery, pathology and scintigraphic results: group I (thyroid extent), group II (locoregional extent), and group III (distant metastatic disease). Clinical parameters, 131I scans, serum thyroglobulin determinations and 131I therapeutic administered doses were evaluated in the follow-up. Some other complementary techniques such as chest X-ray and pulmonary function tests are also described. Scintigraphic absence of thyroid tissue has been observed in 83% of the cases; high thyroglobulin level is still detectable in 34% of the patients as a single evidence of disease, and 21% remain without any abnormal clinical, scintigraphic or analytical findings. Total doses administered have increased in groups I, II and III respectively, and have also been inversely proportional to the extension of lymph node surgery. At present, all the patients are alive and in good general condition. According to the results obtained, we conclude that children and young adults with DTC should undergo periodical 131I therapeutic doses in case of positive scans (once total thyroidectomy has been realized, with or without lymph node resection depending on the extension of disease). In our experience, the use of radioiodine is effective and safe in the follow-up of children and youngs with DTC. PMID- 14534008 TI - [Brain perfusion SPECT in the prognostic assessment of stroke]. AB - PURPOSE: To compare the diagnostic yield of CT scan and perfusion SPECT on admission and its prognostic value in clinical outcome. METHODS: 25 ischemic stroke cases were studied on admission (<24 h) and at 30-60 days by CT scan, 99mTc-HMPAO-SPECT and neurological scales. Infarct size and severity on SPECT were assessed: visually "Total Weighted Score," added value in 22 areas, and by several semiquantitative count-based indices. RESULTS: Sensitivity: the first CT scan was positive in 24% patients, initial SPECT in 75% (73% of pure subcortical infarcts and 91% of those with cortical involvement). Localization: kappa: 0.725 between SPECT findings on admission and those in control-CT at 5 days. Extent and severity: correlations between count-based and visual indices (r: >0.719), the latter correlated slightly better with clinical scales. Both predicted similarly (Rho>0.739) infarct size in CT diagnostic scan. Early Outcome: There were statistical differences between deceased and survivors in SPECT (<24h) indices and CT-infarct size (mean 5 days), but not in neurological scores on admission. Long term Outcome: Correlation of initial SPECT indices with follow-up functional scores (SNSLP, Barthel index; mean 37 days) was only significant for visual SPECT indices (Rho:0.560 to 0.620). Nevertheless the best predictor of functional status on discharge was the Barthel Index on admission. CONCLUSIONS: 1) Early SPECT has good sensitivity and accurate infarct size prediction so it can be a useful tool for deciding thrombolytic therapy; 2) Visual scores perform as well as more complex indices; 3) Infarct volume seems to be a critical determinant in vital outcome; other factors (strategic localization, etc.) might influence long term functional status. PMID- 14534009 TI - [Scintigraphy with thallium chloride in a case of clear cell sarcoma of the tendons and aponeurosis]. AB - Clear cell sarcoma of the soft parts (CCSSP) is a rare and highly malignant tumor. This is a case report of a 31 years old woman who presented with a tumor in the internal face of left thigh. The tumor biopsy was suggestive of a possible malignant tumor of the peripheral nerve sheath (malignant schwannoma). Biochemical analyses, computed tomography and magnetic resonance were performed and reported a sarcoma of soft parts (CCSSP) without abnormal inguinal lymph nodes. With this diagnostic suspicion, the patient was sent to the Nuclear Medicine Service of our center where a 201Thallium scintigraphy study was performed. In this study, it showed the primary tumor together with a hot spot in the homolateral inguinal region, suggestive of the presence of a metastastic lymph node. This finding would change the surgical attitude in the patient, including inguinal lymphadenectomy. The post-surgical histological study confirmed a clear cell sarcoma of soft parts (CCSSP) with a metastatic inguinal lymph node. PMID- 14534010 TI - [Artifactual image by intra-arterial injection of 99mTC-MDP simulating an osteoarticular condition]. PMID- 14534011 TI - [Hiatus hernia mimicking an endothoracic goiter in thyroid scintigraphy]. PMID- 14534012 TI - [Tc99m-HMPAO-labeled leukocyte scan in a case of infected aortic aneurysm]. PMID- 14534014 TI - [Diagnosis and treatment of differentiated thyroid carcinoma]. PMID- 14534016 TI - [Interventional radiology, angioplasty and TIPS in Budd-Chiari syndrome]. AB - INTRODUCTION: Budd-Chiari syndrome consists of complete or partial obstruction of the hepatic veins. Many treatment options are available; new interventional radiology techniques may play an important role as an alternative to surgical shunting. PATIENTS AND METHOD: Of 11 patients diagnosed with Budd-Chiari syndrome, 7 were treated with percutaneous balloon angioplasty or transjugular intrahepatic portosystemic shunt (TIPS) (2 and 5 patients, respectively). The efficacy of both techniques in the treatment of this syndrome was evaluated. RESULTS: Two patients presented significant isolated stenosis of a suprahepatic vein and underwent percutaneous balloon angioplasty. In both patients outcome was favorable with resolution of ascites. Five patients underwent TIPS and four showed clinical improvement with significant reduction in Child-Pugh score (p < 0.05) and resolution of ascites. No shunt malfunction was detected during follow up. One patient showed no improvement after placement of TIPS and died soon after the procedure and a further two died from their underlying disease during follow up. CONCLUSIONS: Treatment of Budd-Chiari syndrome requires multidisciplinary evaluation and should be individualized. In patients with Budd-Chiari syndrome uncontrolled by medical therapy, TIPS may become the decompressive method of choice as an alternative to surgical shunting. Liver transplantation may be reserved to patients in whom these techniques are ineffective. PMID- 14534017 TI - [Interferon, ribavirin and amantadine in prior nonresponders to interferon and ribavirin therapy with chronic hepatitis C (genotype 1)]. AB - OBJECTIVE: Despite advances in the treatment of chronic hepatitis C virus (HCV), the disease persists after treatment with interferon and ribavirin in a large percentage of patients and other therapeutic options are lacking. We investigated the efficacy of retreatment with antiviral therapy including amantadine. EXPERIMENTAL DESIGN: prospective and open pilot study. PATIENTS: Thirty-nine patients with chronic HCV, genotype 1, who were nonresponders to interferon and ribavirin were included. The patients were given repeat treatment with interferon alpha 2A (9 MU/week), ribavirin (1,000-1,200 mg/day) and amantadine (200 mg/day) for 48 weeks. RESULTS: HCV-RNA was undetectable in 5 patients in week 48 (12.8%) and in only 2 patients after 24 weeks of follow-up (5.1% of sustained responses). In patients with basal viremia of < 8 105 U/ml the probability of response at the end of treatment and of sustained response was 26.3 and 10.5%, respectively; in patients with elevated viremias response was 0%. CONCLUSIONS: In patients with chronic HCV genotype 1 without response to interferon and ribavirin, triple antiviral therapy with interferon, ribavirin and amantadine is not useful. PMID- 14534018 TI - [Lactose malabsorption in ulcerative colitis. A case-control study]. AB - BACKGROUND: The role of lactose malabsorption in ulcerative colitis is controversial. The aim of this study was to compare the prevalence of lactose malabsorption in a group of ulcerative colitis patients and a control group and to modify lactose consumption in view of the results. METHODS: Lactose malabsorption was studied using the hydrogen breath test in 52 patients with ulcerative colitis and 34 controls after ingestion of 25 g of lactose. A questionnaire on ingestion of milk products was also administered. RESULTS: Of the 52 patients with ulcerative colitis, 13 (25%) presented lactose malabsorption compared with 11 of the 34 (32%) controls (p = 0.45). Twenty-four patients (46%) had been advised to completely eliminate lactose from their diets. Twenty-seven of the 39 patients without malabsorption had reduced or eliminated lactose consumption after being diagnosed with ulcerative colitis. CONCLUSIONS: No significant differences in the prevalence of lactose malabsorption was found between patients with ulcerative colitis and controls. We believe that systematic elimination of lactose from the diets of these patients is erroneous. In our environment, we recommend the hydrogen breath test only in patients with symptoms of lactose intolerance. PMID- 14534019 TI - [Autoimmune hepatitis and anti-Ro antibodies. Is there a link?]. AB - Antinuclear antibodies (ANA) are considered to be markers of autoimmune disease. Several specific reactivities of ANA are known, among them the Ro (SS-a) complex. Although the presence of anti-Ro antibodies is widely known in systemic connective tissue diseases such as lupus erythematous or Sjogren's syndrome, few studies have attempted to link this finding with autoimmune hepatitis (AIH). We present a case of acute AIH in a 55-year-old woman who tested positive for anti Ro 60 kD antibodies. The Ro (SS-A) antigen is a complex ribonucleoprotein whose structure is still the subject of debate. Two fractions, of 50 and 60 kD, have been identified. A review of the literature revealed frequencies of 21-34% for anti-Ro 52 kD and of 9-13-6% for anti-Ro 60 kD in AIH. Although no relationship has been demonstrated between this complex and the etiopathogenic mechanisms or clinical patterns of AIH, we propose that multicenter investigations with large series should be performed before the possible involvement of anti-Ro antibodies in these diseases is definitively ruled out. PMID- 14534020 TI - [Eosinophilic enteritis as a rare cause of ascites]. AB - Eosinophilic gastroenteritis is an infrequent entity characterized by tissular eosinophilia that can affect different layers of the intestinal wall. This entity can affect any area of the digestive apparatus from the esophagus to the rectum. Clinical manifestations depend on the affected layers and range from barely perceptible symptoms to intestinal obstruction or ascites. We present the case of an 18-year-old woman who showed abdominal ascites as a rare form of presentation with difficult differential diagnosis with peritoneal carcinomatosis. PMID- 14534021 TI - [Ileo-ileal and ileocecal invagination due to intestinal lipomatosis]. AB - Intestinal lipomatosis is a rare entity and few cases have been reported in the literature. The condition is usually asymptomatic. Symptomatic cases usually present as obstruction or, less frequently, as bleeding. Intestinal barium studies, ultrasonography and computed tomography are useful diagnostic techniques. We present the case of a 47-year-old man with no relevant medical history who presented with intestinal obstruction of several months' duration. Complementary investigations yielded a diagnosis of intestinal obstruction due to ileocecal invagination secondary to endoluminal tumors of the ileum. Surgery and pathological analysis revealed the latter to be intestinal lipomatosis. This rare clinical entity has been associated with diverticulosis and intestinal volvulus. PMID- 14534022 TI - [Clinical strategy to prevent the gastrointestinal adverse effects of nonsteroidal anti-inflammatory agents]. PMID- 14534023 TI - [Role of dietary lipids in the mechanisms of inflammation, proliferation, differentiation and cell death in the gastrointestinal tract]. PMID- 14534024 TI - [Combined treatments for esophageal varices]. PMID- 14534025 TI - [Nodular regenerative hyperplasia and common variable immunodeficiency]. PMID- 14534029 TI - Differences in adolescent smoker and nonsmoker perceptions of strategies that would help an adolescent quit smoking. AB - This study assessed adolescent smoker and nonsmoker perceptions of strategies that would help an adolescent smoker in his or her attempt to stop smoking. Surveys were distributed primarily in the schools at 4 geographic and ethnically diverse study sites. Respondents were 965 adolescents (49% female; 46% minority). Current smokers (n = 232) were asked to rate the extent to which they agreed or disagreed that supportive behaviors of friends and family, quitting strategies, or learning about quitting strategies would be helpful if they decided to quit. Nonsmokers (n = 733) were asked to indicate the degree to which they agreed or disagreed that these behaviors and strategies would be helpful if a friend decided to quit. Responses to each of the 33 attitude items were rated on a 5 point scale ranging from strongly disagree to strongly agree. Marked differences were observed between smokers and nonsmokers in the level of agreement on each item. In general, smokers reported far less enthusiasm for cessation strategies than nonsmokers. After adjusting for gender, age, and other covariates, smoking status was the strongest independent predictor of the number of items endorsed as agree or strongly agree. The results have implications for the design of peer based and other interventions for adolescent smokers. PMID- 14534028 TI - Methodological challenges in research on sexual risk behavior: II. Accuracy of self-reports. AB - Assessing sexual behavior with self-report is essential to research on a variety of health topics, including pregnancy and infertility, sexually transmitted infections, and sexual health and functioning. Recent methodological research has provided new insights regarding the accuracy of self-reports of sexual behavior. We review these studies, paying particular attention to a promising new development: the use of computer-assisted assessments. The collection of sexual risk behavior data with computers has increased dramatically in recent years, but little is known about the accuracy of such assessments. We summarize the evidence, discuss methodological issues that arise in studies evaluating the accuracy of self-reports, and offer recommendations for future research. PMID- 14534027 TI - Methodological challenges in research on sexual risk behavior: I. Item content, scaling, and data analytical options. AB - Investigation of sexual behavior involves many challenges, including how to assess sexual behavior and how to analyze the resulting data. Sexual behavior can be assessed using absolute frequency measures (also known as counts) or with relative frequency measures (e.g., rating scales that range from never to always). We discuss these 2 assessment approaches in the context of research on HIV risk behavior. We conclude that these 2 approaches yield nonredundant information and, more important, that only data yielding information about the absolute frequency of risk behavior have the potential to serve as valid indicators of HIV contraction risk. However, analyses of count data may be challenging because of non-normal distributions with many outliers. Therefore, we identify new and powerful data analytical solutions that have been developed recently to analyze count data and discuss limitations of a commonly applied method (viz., analysis of covariance using baseline scores as covariates). PMID- 14534030 TI - Autonomic responses to psychological stress: the influence of menopausal status. AB - Cardiovascular disease is the leading killer of women in developed countries. Menopause, stress, and lack of social support may contribute to the increased risk of heart disease morbidity and mortality in women. This study examined the effects of psychological stress and social support on autonomic nervous system control of the heart in 18 pre- and 34 postmenopausal women. Autonomic activity, as indexed by high-frequency heart rate variability and pre-ejection period, was assessed at rest and during a public speech task. Social support was determined using the Berkman Social Support Inventory. Postmenopausal women had higher heart rate (F = 4.4, p pound.04) and less parasympathetic activity (F = 11.9, p pound.001) compared with premenopausal women at rest. In response to stress, sympathetic nervous system activity increased in postmemopausal women (F = 6.1, p pound.02); however, this effect was no longer significant when age was used as a covariate in the analysis. Social support did not significantly affect measures of autonomic activity in either the pre- or postmenopausal women. Postmenopausal women have greater sympathetic and less parasympathetic activity than premenopausal women, which may account for their increased risk of coronary artery disease. PMID- 14534031 TI - Signaling does not adequately improve diary compliance. AB - HYPOTHESIS: Compliance with a paper diary protocol would be improved by using auditory signaling. BACKGROUND: Prior research has demonstrated that compliance with the reporting schedule in paper diary protocols is poor. METHODS: Adults with chronic pain (N = 27) were recruited from the community to participate in a 24-day experience sampling protocol of 3 pain assessments per day (10:00 a.m., 4:00 p.m., 8:00 p.m.). Diaries were instrumented to record openings and closings, thereby permitting determination of date and time when the participant could have made diary entries. Participants were signaled with a programmed wristwatch at the onset of each 30-min assessment window. Two compliance windows were defined: /+ 15 min and -/+ 45 min of the targeted assessment time. RESULTS: Self-reported compliance based on participants' paper diaries was 85% and 91% for the 30- and 90-min windows. Verified compliance was 29% and 39% for the two windows. Signaling produced a significant increment in verified compliance when compared with an identical trial without signaling. A significant eroding of verified compliance was observed across the 3 weeks of the study. CONCLUSIONS: Self-report dating of diary entries may be misleading investigators about compliance with diary protocols. Although auditory signaling enhances compliance, the result is still unsatisfactory. PMID- 14534032 TI - The role of postpartum weight retention in obesity among women: a review of the evidence. AB - Young, adult women appear to be at increased risk for substantial weight gain. Pregnancy has frequently been cited as a contributor to overweight in women. This article reviews the literature examining the role of pregnancy in the development of overweight. Average postpartum weight retention ranges from 0.5 to 3 kg; however, postpartum weight retention appears to be highly variable among women, with some women retaining as much as 17.7 kg. Excessive postpartum weight retention seems to be especially prevalent among minority women. Factors such as prepregnancy weight and excessive gestational weight gain have the strongest support as risk factors for postpartum weight retention and could guide targeted intervention efforts. However, there are few controlled studies of behavioral interventions to prevent substantial pregnancy-related weight gain or postpartum weight retention. Weight loss methods successful in promoting weight control in other populations would likely also be effective with pregnant or postpartum women, although modifications for the needs of mothers may be required. Particular attention to the needs of minority mothers is warranted given the likelihood of greater weight retention in this group. PMID- 14534033 TI - Drug effects on calcium homeostasis in mouse CA1 hippocampal neurons. AB - Voltage-dependent Ca2+ channels (VDCC) are important in control of neuronal excitability, synaptic transmission, and many other cellular process. Even the slightest alteration in Ca2+ currents can have a considerable impact on the neuronal function. However, it is still unknown whether Ca2+ currents are affected by neurotoxic drugs such as lead, cobalt, zinc, cadmium, thallium, lanthanum, and aluminum. We have characterized the effects of neurotoxic drugs on Ca2+ homeostasis in CA1 hippocampal C57BL mice. Fura 2-AM fluorescence photometry was used to measure intracellular Ca2+ concentration ([Ca2+]i) in the presence and absence of neurotoxic drugs (10 microM) in response to KCl application. The peak [Ca2+]i due to KCl application was reduced in the presence of lead (60%), cobalt (35%), zinc (62%), cadmium (71%), thallium (27%), and lanthanum (66%). By contrast, in the presence of aluminum the peak [Ca2+]i was either increased (46%) or it was not affected. These results indicate that neurotoxic drugs could block the entry of calcium into CA1 neurons via VDCC. PMID- 14534034 TI - Relearning of locomotion in injured spinal cord: new directions for rehabilitation programs. AB - Locomotor activity-based rehabilitation programs seem to bring encouraging recovery to incomplete spinal cord injury paralytics. But in "complete" injuries recovery is long delayed and in small increments. Developmental and computer neural networks studies show that motor relearning requires the participation of redundant numbers of and activity-dependent competition of synapses among spinal interneurons. The present programs do not meet this prerequisite. They focus mainly on the mechanical (retraining) part while the neural (relearning) part plays a rather passive role. Here we suggest how retraining can be integrated with relearning by inductive lability procedures (Krishnan, 2003a, 2003b) with the objective of enhancing and hastening recovery. PMID- 14534035 TI - Electrophysiological evaluation of genito-sphincteric dysfunction in multiple system atrophy. AB - Thirteen patients with multiple system atrophy underwent multimodality neurophysiological evaluation, including sphincteric needle electromyography (EMG), sacral reflexes, pudendal nerve terminal latency, pudendal (PSEPs) and tibialis posterior nerve somatosensory evoked potentials (TPSEPs), and perineal motor evoked potentials (PMEPs). EMG revealed denervation or neurogenic changes, with reduction in spontaneous tonic activity at rest and abnormal pudendal nerve terminal latency in 10 patients (76.9%); anal reflex was delayed in 7 patients (53.8%). TPSEPs scalp responses were clearly abnormal in 4 patients (30.7%), whereas PSEPs exhibited changes in 9 (69.2%): in 6 patients responses were delayed at lumbar level (46.2%), and in 5 over the scalp (38.4%). PMEPs showed an increase in latency with a mild prolongation of central motor conduction time (CMCT) in 2 cases (15.3%); 1 patient had prolonged latencies following both cortical and sacral stimulation, but a normal CMCT. Even if diagnostic yield is not improved using these investigations they provide evidence of multiple lesion sites other than Onuf's nucleus. PMID- 14534036 TI - Prevalence of left-handedness among patients with different respiratory diseases. AB - We investigated the prevalence of left-handedness among patients with different respiratory diseases. Hand preference was assessed using the modified Edinburg Handedness Inventory. There was a statistically significant decline in left handedness with shortened life span in patients. Difference among the rates of left-handedness in different pulmonary diseases was significant. The rate of left handedness was lowest in patients with chronic obstructive pulmonary disease (2.14%) and pneumonia (2.04%) and highest in patients with asthma (12.96%). The results suggest that left-handed subjects may be protected from certain pulmonary diseases while being more susceptible to others. PMID- 14534037 TI - Evidence for a role of dopaminergic mechanisms in the immunostimulating effect of mu-opioid receptor agonist DAGO. AB - The present study demonstrates that: (1) activation of micro -opioid receptors by systemic administration of a highly selective agonist DAGO (100 microg/kg) results in a significant increase in the number of plaque- and rosette-forming cells in the spleen of CBA mice as well as Wistar rats on the 5th day following sheep red blood cells (5 x 10(8)) immunization, (2) the immunostimulatory effect of DAGO is mediated by central mechanisms including the hypothalamus-hypophysis complex; (3) the postsynaptic dopamine (DA) receptors of D2 type are involved in the DAGO-induced immunostimulation since the combined treatment of animals with haloperidol (2 mg/kg), a blocker of DA D2 receptors, and DAGO abolished this effect; (4) the nuclei caudatus and accumbens of the nigrostriatal and mesolimbic DAergic systems, respectively, are implicated in the immune response stimulation caused by DAGO. PMID- 14534039 TI - Serotonin reduces potassium current in rutabaga and wild-type Drosophila neurons. AB - The Drosophila learning mutant rutabaga is defective in short-term memory and has a reduced intracellular cyclic adenosine monophosphate (cAMP) concentration. The delayed-rectifier potassium current (IKDR) was measured from cultured (2 days) wild-type and rutabaga neurons. IKDR was smaller in rutabaga neurons (382 +/- 41 pA) than in wild-type neurons (542 +/- 33 pA). IKDR was measured from neurons before and after addition of serotonin to the external solution. IKDR was reduced by serotonin in wild type (decreasing 37 +/- 7%) and rutabaga (decreasing 33 +/- 6%) neurons (single-cell studies). In the presence of serotonin, IKDR was smaller in rutabaga (218 +/- 24 pA) than in wild-type (426 +/- 35 pA) neurons (population studies). These results indicate that serotonin has affected IKDR so that the inherent difference between the two genotypes was preserved. PMID- 14534038 TI - Hypothalamic digoxin, hemispheric chemical dominance, and peptic ulcer disease. AB - The isoprenoid pathway produces three key metabolites--endogenous digoxin-like factor (EDLF) (membrane sodium-potassium ATPase inhibitor and regulator of neurotransmitter transport), ubiquinone (free radical scavenger), and dolichol (regulator of glycoconjugate metabolism). The pathway was assessed in peptic ulcer and acid peptic disease and its relation to hemispheric dominance studied. The activity of HMG CoA reductase, serum levels of EDLF, magnesium, tryptophan catabolites, and tyrosine catabolites were measured in acid peptic disease, right hemispheric dominant, left hemispheric dominant, and bihemispheric dominant individuals. All the patients with peptic ulcer disease were right-handed/left hemispheric dominant by the dichotic listening test. The pathway was upregulated with increased EDLF synthesis in peptic ulcer disease (PUD). There was increase in tryptophan catabolites and reduction in tyrosine catabolites in these patients. The ubiquinone levels were low and free radical production increased. Dolichol and glycoconjugate levels were increased and lysosomal stability reduced in patients with acid peptic disease (APD). There was increase in cholesterol:phospholipid ratio with decreased glyco conjugate levels in membranes of patients with PUD. Acid peptic disease represents an elevated EDLF state which can modulate gastric acid secretion and the structure of the gastric mucous barrier. It can also lead to persistence of Helicobacter pylori infection. The biochemical pattern obtained in peptic ulcer disease is similar to those obtained in left-handed/right hemispheric chemically dominant individuals. But all the patients with peptic ulcer disease were right-handed/left hemispheric dominant by the dichotic listen ing test. Hemispheric chemical dominance has no correlation with handedness or the dichotic listening test. Peptic ulcer disease occurs in right hemispheric chemically dominant individuals and is a reflection of altered brain function. PMID- 14534040 TI - Hypothalamic digoxin, hemispheric chemical dominance, and interstitial lung disease. AB - The isoprenoid pathway produces three key metabolites--endogenous digoxin, dolichol, and ubiquinone. This was assessed in patients with idiopathic pulmonary fibrosis and in individuals of differing hemispheric dominance to find out the role of hemispheric dominance in the pathogenesis of idiopathic pulmonary fibrosis. All 15 cases of interstitial lung disease were right-handed/left hemispheric dominant by the dichotic listening test. The isoprenoidal metabolites -digoxin, dolichol, and ubiquinone, RBC membrane Na(+)-K+ ATPase activity, serum magnesium, tyrosine/tryptophan catabolic patterns, free radical metabolism, glycoconjugate metabolism, and RBC membrane composition--were assessed in idiopathic pulmonary fibrosis as well as in individuals with differing hemispheric dominance. In patients with idiopathic pulmonary fibrosis there was elevated digoxin synthesis, increased dolichol and glycoconjugate levels, and low ubiquinone and elevated free radical levels. There was also an increase in tryptophan catabolites and a reduction in tyrosine catabolites. There was an increase in cholesterol phospholipid ratio and a reduction in glycoconjugate level of RBC membrane in patients with idiopathic pulmonary fibrosis. Isoprenoid pathway dysfunction con tributes to the pathogenesis of idiopathic pulmonary fibrosis. The biochemical patterns obtained in interstitial lung disease are similar to those obtained in left-handed/right hemispheric chemically dominant individuals by the dichotic listening test. However, all the patients with interstitial lung disease were right-handed/left hemispheric dominant by the dichotic listening test. Hemispheric chemical dominance has no correlation with handedness or the dichotic listening test. Interstitial lung disease occurs in right hemispheric chemically dominant individuals and is a reflection of altered brain function. PMID- 14534041 TI - Systematic rules underlying spectral pattern variability: experimental results and a review of the evidence. AB - On the basis of three different experiments: oddball task (visual, auditory, and audio-visual stimuli), modified Sternberg's, and multistage memory tasks, it was shown that: a) there was not a single typical spectral pattern type that would characterize the majority of the trials; b) the total number of the different spectral pattern types was limited; c) different spectral pattern types had different importance to the brain--their occurrence was more or less probable; d) the total number and the number of the most probable spectral pattern types were dependent on the functional brain state; e) actual spectral pattern of variability during rest with closed eyes was relatively high (around 65% from the maximum possible rate), but significantly less than stochastic spectral pattern variability. It is suggested that identical sensory events can potentially trigger a limited number of several different alternative reaction patterns in EEG/MEG, depending on the situational context. PMID- 14534042 TI - Research and practice--are we giving children the best treatment? PMID- 14534043 TI - Dysarthria and dysphagia as long-term sequelae in a child treated for posterior fossa tumour. AB - The current case report provides a comprehensive description of the persistent dysarthria and dysphagia evident in a 7.5 year old child treated for recurrent posterior fossa tumour (PFT). AC was assessed on a comprehensive perceptual and instrumental test battery incorporating all components of the speech production system (respiration, phonation, resonance, articulation and prosody) 2 years and 4 months following completion of her treatment. The nature of her swallowing impairment was investigated through the use of videofluoroscopic evaluation of swallowing (VFS). A mild dysarthria with ataxic and LMN components was identified, although overall speech intelligibility was not affected. A moderate dysphagia was also identified with impairment in all three phases of the swallowing process; oral preparatory, oral and pharyngeal. Dysarthria and dysphagia as persistent sequelae in children treated for PFT have implications for the long-term management of these children. The need for appropriate treatment regimes, as well as pre-surgical counselling regarding dysarthria and dysphagia as possible outcomes following surgery are highlighted. PMID- 14534044 TI - Development of a disease-specific disability instrument for Pompe disease. AB - PURPOSE: The purpose of this study was to modify the Paediatric Evaluation of Disability Inventory (PEDI) to create a Pompe disease-specific disability instrument for use in clinical trials and natural history studies. METHODS: PEDI item content was revised to include self-care and mobility items appropriate for children and youth with Pompe disease. Data were collected on 30 individuals with Pompe disease (mean age 7.7+/-5.6 years; range 0.4-22.1 years) by parent proxy through telephone interviews. New items were merged with original PEDI items using Rasch rating scale methods. RESULTS: The Pompe-PEDI extended the content range and scoring precision of the original PEDI. Construct validity was demonstrated and test-re-test reliability was excellent. CONCLUSIONS: The Pompe PEDI is a reliable and valid instrument to assess and monitor the functional changes of children and youth with Pompe disease. PMID- 14534045 TI - Evidence-based approach of the use of Botulinum toxin type A (BTX) in cerebral palsy. AB - A systematic review with the Sachett model of evidence-based medicine of the use of Botulinum toxin type A (BTX) for intervention in children with Cerebral Palsy (CP) is highlighted. Currently, the evidence showed that BTX is useful for treating pes equinus due to spasticity of the gastrocnemius-soleus muscles. However, careful patient selection and goals of treatment have to be addressed. More multi-centre clinical trials with standardized protocols are needed before widespread recommendation of the use of BTX in treating spasticity in CP can be made. PMID- 14534046 TI - An assessment of the impact of thimerosal on childhood neurodevelopmental disorders. AB - The prevalence of autism in the US has risen from 1 in approximately 2500 in the mid-1980s to 1 in approximately 300 children in the mid-1990s. The purpose of this study was to evaluate whether mercury from thimerosal in childhood vaccines contributed to neurodevelopmental disorders. Neurodevelopmental disorder dose response curves for increasing mercury doses of thimerosal in childhood vaccines were determined based upon examination of the Vaccine Adverse Events Reporting System (VAERS) database and the 2001 US' Department of Education Report. The instantaneous dosage of mercury children received in comparison to the Food and Drug Administration (FDA)'s maximum permissible dose for the oral ingestion of methylmercury was also determined. The dose-response curves showed increases in odds ratios of neurodevelopmental disorders from both the VAERS and US Department of Education data closely linearly correlated with increasing doses of mercury from thimerosal-containing childhood vaccines and that for overall odds ratios statistical significance was achieved. Similar slopes and linear regression coefficients for autism odds ratios in VAERS and the US Department of Education data help to mutually validate each other. Controls employed in the VAERS and US Department of Education data showed minimal biases. The evidence presented here shows that the occurrence of neurodevelopmental disorders following thimerosal containing childhood vaccines does not appear to be coincidental. PMID- 14534047 TI - Age and recovery from brain injury: legal opinions, clinical beliefs and experimental evidence. AB - Despite considerable scientific evidence to the contrary, many medical practitioners maintain that children recover from brain injury better than adults. This belief, which is commonly referred to as the "Kennard Principle", has important ramifications for personal injury compensation claims in which the amount of financial damages claimed is partly based on medical experts' prognoses for recovery and long-term outcome. The present study investigated whether legal practitioners' beliefs are consistent with those of medical practitioners. Lawyers were asked to estimate their confidence in consultant neurologists' estimates of recovery in four clinically-based but fictitious case studies which differed only in the reported age of the patient. The lawyers showed more confidence in estimates which coincided with the Kennard Principle than those which did not. These results support previous findings in showing widespread belief that "younger is better" in recovery from brain injury. In consequence, it is likely that financial compensation for children with brain injury is currently being underestimated in litigation, thereby prejudicing the long-term outcome of the child. PMID- 14534048 TI - Incidence of surgery in conservatively treated patients with scoliosis. AB - The effectiveness of bracing and the effectiveness of conservative treatment of scoliosis as a whole is widely denied and rejected. We therefore carried out the present study in order to establish whether the work involved in the conservative treatment of scoliosis is reasonable and worthwhile, including as it does the education of patients, techniques of brace moulding, time-consuming follow-ups and the strain put on patients by the treatment itself. STUDY DESIGN: A retrospective analysis of the incidence of surgery in patients with scoliosis presenting every kind of aetiology. To improve comparison with data already published by other centres, this group of patients was matched with the study design of the control group. MATERIALS AND METHODS: Since 1992, in addition to the intensive inpatient rehabilitation programme developed at our centre, we have also been practising moulding techniques according to Cheneau in order to construct scoliosis orthoses of high correction. For the purposes of this study we chose a number of scoliosis patients from our data bank who had had conservative treatment at our centre between 1993 and 1996. All the patients, like those of the untreated control group, were at least 15 years of age at the time they were last investigated or questioned. The incidence of surgery in our group was compared statistically with data published by other centres. RESULTS: Three hundred and forty-three patients (females only) with a curvature of 33.4 degrees (SD = 18.9) were included in our study and could be followed up under the described pre-conditions. Forty-one patients (11.95%) had had surgery. In patients with adolescent idiopathic scoliosis, we found an incidence of surgery of only 7.3% compared with 28% in the control group with the same diagnosis. Statistically, therefore, the incidence of surgery in our group was highly significantly lower than the incidence of surgery in the control group from Ireland and other centres described in literature. CONCLUSIONS: The conservative treatment of scoliosis (which includes outpatient physiotherapy, inpatient intensive scoliosis rehabilitation (SIR) and high-correction-effect bracing based on plastercasting) shows an obvious reduction in the rate of surgery in patients with idiopathic scoliosis compared with untreated scoliosis patients. Thus it can be seen that the conservative treatment of scoliosis may reduce the incidence of surgery, which not only causes considerable medical expenses but also carries with it the risk of repeated operations and further complications. In our study, we also found highly significant differences by comparison with the work of a bracing centre in the USA. Therefore, the quality of the bracing itself and the effects of primary correction must receive greater consideration in future scientific studies on the subject of "conservative" treatment of scoliosis. PMID- 14534049 TI - Executive functions and rehabilitation of the integrated self: a commentary on Ylvisaker and Feeney. AB - This commentary is a response to Ylvisaker and Feeney's paper "Executive functions, self-regulation and learned optimism in paediatric rehabilitation: a review and suggestions for intervention". Drawing on a diverse literature, the authors provide a broad empirical basis for a pragmatic approach to rehabilitation for children. Their contribution is critically reviewed in terms of the validity of their central thesis and the likely impact of their proposals. Although theoretically ambiguous in places and lacking the benefit of a truly developmental framework for intervention, their proposals, nevertheless, represent a coherent heuristic framework for teachers, clinicians and parents that has corollary implications for how paediatric rehabilitation is funded. PMID- 14534057 TI - Quality of life of mothers having nosocomially HIV-infected children in Russia. AB - BACKGROUND: 274 children were HIV-infected during 1988-89 in hospitals in the South of Russia during treatment of heavy pathology requiring an intensive therapy and catheterization of large venous vessels. Today only 140 children, receiving HAART now, remain alive. PATIENTS AND METHODS: In 1988 - 89 mothers having nosocomially HIV-infected children were tested on a 'WHOQOL-100' technique (Russian version). FINDINGS: Though the majority of mothers estimated the basic domains of quality of life (QoL) as average and good, the significant part considers their QoL as bad (in physical health, 22.7%, in psychological health, 20.4%, in environment, 25%, in social relationship, 11.4%). The least data of the test were in facets of financial resources, medical and social help and 47.7% consider a general QoL and condition of health as bad, 54.5% marks a lack of positive emotions, and 52.3% had plenty of negative emotions. 45.5% of mothers marked unsatisfactory opportunities for rest and entertainments, 31.8% of those interrogated had problems with an environment of a house. For the most part, mothers considered their health as 'neither bad, nor good' (63.9%), as 'basically bad' (9.1%), and 22.7% as 'basically good'. The basic problems are 'heart problems' (38.6%), 'increased blood pressure' (25%), 'chronic nervous/emotional problems' (36.4%), 'depression' (31.8%). The absence of problems with health was noted by only 9.1% of those surveyed. Overall, 54.5% of mothers considered themselves as patients, and 22.7% as requiring medical treatment 'rather urgently'. At the same time, only 11.4% had received stationary treatment 'in the last 2 weeks', and 15.9%, any out-patient treatment. INTERPRETATION: The problems of QoL and of psychosocial rehabilitation of mothers having nosocomially HIV infected children are discussed. PMID- 14534058 TI - Long-term disabilities and handicaps following sports injuries: outcome after outpatient treatment. AB - PURPOSE: The aim was to investigate whether long-term disabilities and handicaps arise from a sports injury requiring outpatient treatment and to identify the potential risk factors. METHOD: A representative sample was taken from a population of patients treated as outpatients due to a sports injury. The selected patients were sent a questionnaire, 2-5 years after the injury. RESULT: Thirty-nine percent of the patients studied were unable to work for up to 1 month after the injury, 19% were not able to work for up to 3 months and another 5% could not work for a maximum of 8 months. Participation in sporting activities was hampered for up to 1 year in 76% of the patients and 11% had not resumed sports participation at all. In addition, 20% of the population stated that they still suffered from disabilities and handicaps following the sports injury. The outcome of the SIP68 underlines these results. Nine percent of the patients had a sumscore larger than 0. The variables which could be identified as risk factors were the body region: knee and sex: female. CONCLUSIONS: Sports injuries requiring outpatient treatment can lead to long-term disabilities and handicaps, especially in patients with knee injuries and injuries in women. On average these consequences are less severe than those associated with inpatients; however, this finding is of great value since the number of outpatients is much higher than in patients admitted to a hospital. PMID- 14534059 TI - Characteristics of disability and handicap among Toxic Oil Syndrome (TOS) cohort patients: a cross-sectional study, 17 years after the original food intoxication. AB - PURPOSE: In 1981, a progressive multi-systemic disease called Toxic Oil Syndrome (TOS) appeared in Spain as an epidemic that affected 20,000 people. The International Classification of Impairments, Disabilities and Handicaps (ICIDH) was chosen to characterize the health status of patients more severely affected by TOS. METHODS: A random sample of 292 with permanent disability was selected. Disability was assessed with a questionnaire based on ICIDH and the Stanford Health Assessment Questionnaire. Handicap was measured using London Handicap Scale. Distributions of the proportions and 95% confidence intervals for disabilities, handicaps were calculated and stratified by dimensions, age and sex. The chi2 test was used for inter-group comparisons. RESULTS: Two hundred and fourteen patients were interviewed. Mobility-related and behaviour disabilities were most prevalent. Disability rose with age and was higher among women, except for behaviour disabilities which were more frequent in young men. Mean handicap score was 78.0 +/- 12.7. Handicap dimensions most affected were physical independence and economic self-sufficiency. CONCLUSIONS: The health profile of the population hardest hit by TOS is characterized by the presence of important functional and psychosocial disabilities that limit performance of daily living activities and social role, and are in accord with the handicap that such persons suffer. PMID- 14534060 TI - Experiencing the loss of mobility: perspectives of older adults with MS. AB - PURPOSE: The purpose of this study was to develop an understanding the experience and meaning of mobility loss among older adults with multiple sclerosis (MS). This work is part of a larger study on the health concerns and service needs of older adults with MS. METHODS: Twenty-seven people with MS (mean age = 62 years, range = 55-82) were interviewed using a phenomenological approach. RESULTS: Thematic analysis found that three factors contributed to participants' experience of mobility as a person with MS: reality of having MS, mobility needs, and contextual factors. Participants discussed how the reality of MS meant variability in their ability to get around and the importance of trying to remain in control over their mobility experiences. The experience of mobility lead participants to mourn losses, take action, and contemplate their futures. They expressed concerns about the continual declines in their mobility, losing independence, becoming a burden on caregivers, and the potential for having to move into a nursing home. CONCLUSIONS: Findings provide insight into the experience of mobility loss among older adults with MS, and point to potential strategies that can be used to help people cope with and adapt to these losses. PMID- 14534061 TI - Determinants of handgrip strength in free-living elderly at risk of malnutrition. AB - PURPOSE: Handgrip strength is necessary for performing activities of daily living, which, in turn, are required to maintain functional autonomy. The purpose of this study was to determine anthropometric and personal factors that affect handgrip strength in a group of free-living elderly at risk of malnutrition. METHOD: The factors associated with handgrip strength (Pearson r, t-test, alpha = 0.10) were entered in a multiple linear regression model (n = 166) to identify the independent prediction factors. Reliability of the model was verified with a sub-group of 65 subjects randomly selected from the initial sample. RESULTS: Both groups were statistically similar regarding all factors studied even though the validation group (n = 65) had more men (32%) compared to the development group (n = 166; 22%). Bivariate analyses showed that handgrip strength was associated with sex, age, pain, hand circumference, and waist-hip ratio. Multiple linear regression analysis identified age, pain, and sex as independent determinants of handgrip strength (R2 = 0.16). This model predicted handgrip strength better in the validation group (R2 = 0.29). CONCLUSIONS: As previously shown, women have a smaller handgrip strength than men. In addition, handgrip strength decreases with increasing age and the presence of pain. Emphasis should be placed on the importance of strategies to relieve pain in the free-living frail elderly population. PMID- 14534063 TI - Should coronary artery revascularization strategies be different for men and women? PMID- 14534062 TI - The environmental barriers of children with Muscular Dystrophies and its effect on mother's low back pain. AB - PURPOSE: The aim of this study is to determine the architectural barriers of children with Muscular Dystrophy at home according to their diminished functional capacity and its effect on their mother's back problem. METHOD: Twenty children with Muscular Dystrophy (11.1 +/- 4.14 years) were evaluated with Brooke Scale and WeeFIM. Quebec and Oswestry Disability Scales were used for determining physical disability of their mothers (38.65 +/- 5.91 years). The architectural barriers have been detected with attention to standard criterias of International Organizations for all countries around the world. RESULTS: The heights of thresholds, switches of wall plugs, shelves, cupboards, mirrors, hangers, arm of windows and siphons were found to be higher than standard values. The relation between back disability questionnaire results of mothers and dependency level of children on Activity of Daily Living has showed negative correlation (p < 0.05). CONCLUSIONS: The protective back health programmes and home arrangements that are suitable for disabled children must be added to the neuromuscular rehabilitation programmes. PMID- 14534064 TI - Glibenclamide blunts coronary flow reserve induced by adenosine and dipyridamole. AB - OBJECTIVE: A reduced coronary flow reserve is considered indicative of significant coronary stenosis. As experimental data suggest that adenosine and dipyridamole induce vasodilatation by opening of ATP-sensitive potassium channels, we sought to determine the effect of glibenclamide, an antidiabetic blocker of ATP-sensitive potassium channels, on adenosine- and dipyridamole induced coronary flow reserve. DESIGN: Coronary flow velocities were measured in 15 pigs using a Doppler flow wire. The effect of increasing glibenclamide concentrations (0.1-10 microM) on adenosine-induced coronary flow reserve was examined in five animals. Ten pigs served as time controls. The time controls were subsequently treated by 3 microM glibenclamide (n = 5) or corresponding vehicle (n = 5) and the flow response to 0.56 mg/kg dipyridamole determined. RESULTS: Glibenclamide elicited a concentration-dependent inhibition of adenosine induced coronary flow reserve, reaching significance at glibenclamide concentrations of 3 and 10 microM. The coronary flow reserve stimulated by dipyridamole was reduced significantly by 3 microM glibenclamide. CONCLUSION: Glibenclamide blunts coronary flow reserve stimulated by adenosine and dipyridamole. This interaction may have clinical implications in diabetics undergoing adenosine- or dipyridamole-dependent diagnostic procedures. PMID- 14534065 TI - Systolic long axis function of the left ventricle. Global and regional information. AB - OBJECTIVE: To compare global systolic measurements of mitral annular motion by M mode and tissue velocity time integral, and annular velocity by pulsed and colour Doppler for precision and bias. Secondly, to compare the ability of annular motion to identify regional dysfunction with segmental analysis by strain rate imaging. DESIGN: Nineteen normal subjects and 19 patients with myocardial infarction were studied with echocardiography. RESULTS: There were significant correlations between ejection fraction (EF) and annular motion/velocity by all methods, ranging from 60 to 80%. Measurements had 95% limits of agreement intervals between 7.7 and 15.6 mm for annulus excursion and 8.8 cm/s for annulus velocities with biases between methods of 0.7-1.9 mm and 2.6 cm/s. Annular motion and velocity were reduced in the patients compared with the control group, but were depressed at all points so the infarcted region could not be identified. Only segmental analysis could identify the region of dyssynergy. CONCLUSION: Annular motion and velocity measure global function, but have high variability and measurements are method dependent. Only segmental analysis can identify regional dyssynergy. This is possible with strain rate imaging, but the precision is still too low for clinical use. PMID- 14534066 TI - Assessment of left ventricular function from M-mode measurement of circumflex artery motion recorded by coronary angiography. AB - OBJECTIVE: To evaluate the usefulness of M-mode measurement of circumflex artery motion (CAM) for assessment of left ventricular (LV) function. DESIGN: Seventy two patients referred for coronary angiography and LV angiography were included. Ejection fraction (EF) was calculated from LV angiography and systolic and diastolic parameters of CAM were measured by M-mode from coronary angiography. Twenty-three patients, examined by echocardiography of mitral annulus motion (MAM) within 24 h before the angiographic examination, formed a subgroup for comparison between angiographic M-mode of CAM and echocardiographic M-mode of MAM. RESULTS: In addition to previous reported CAM amplitude and longitudinal fractional shortening (FSL) the maximal systolic velocity of CAM can be reliably recorded by M-mode. The diastolic indices, atrial contribution to the total amplitude and maximal early and late diastolic velocities, are also well monitored by M-mode of CAM in comparison with echocardiographic MAM. CONCLUSION: LV systolic and diastolic function can be assessed by M-mode of CAM. PMID- 14534067 TI - Optimal timing of aortic valve replacement for aortic stenosis--are we operating late? AB - OBJECTIVE: To evaluate the adherence to current guidelines for surgery in patients with aortic valve stenosis. DESIGN: From 1 January 1997 to 31 May 1999, 99 patients were accepted for aortic valve surgery with preserved left ventricular function and normal coronary angiogram. On admission for operation, 20 patients were evaluated regarding symptoms, exercise capacity, and left ventricular morphology and function. RESULTS: There were 14 men and 6 women, mean age 64.3 years. Years from symptom onset varied from 2.1 to 3.2. Dyspnoea was the most common limiting symptom. Thirty per cent of the patients were classified as NYHA IIIB. Physical capacity was reduced to 79% of the expected. Left ventricular hypertrophy was present in 14/20 patients. Left ventricular systolic function was reduced with mean ejection fraction of 0.46. Diastolic dysfunction (E/A ratio <1) was present in 12 patients. CONCLUSION: Many patients accepted for aortic valve replacement due to aortic stenosis show advanced disease and are referred for surgery later in the disease process than is recommended in the current guidelines. PMID- 14534068 TI - Intramyocardial partial oxygen pressure in patients undergoing transmyocardial laser revascularization and bypass surgery. AB - OBJECTIVE: Numerous investigations could not clarify the exact mechanism of transmyocardial laser revascularization (TMLR). The aim of this study was to investigate, whether TMLR leads to an increase of myocardial oxygenation in comparison to patients undergoing coronary artery bypass grafting (CABG). DESIGN: Twelve patients (TMLR group) underwent TMLR alone with an 800 W CO2 laser through a left anterior thoracotomy. Seventeen patients (CABG group) underwent standard CABG. Myocardial oxygenation was determined by measuring intramyocardial partial oxygen pressure (ptiO2 ). PtiO2 was measured online and mean values at 1, 24, 32, and 48 h postoperatively were compared with baseline before intervention. Parameters influencing ptiO2 (arterial pO2, hemodynamic parameters, hemoglobin) were recorded. RESULTS: Mean baseline ptiO2 was significantly lower in the TMLR group compared with the CABG group (p < 0.05). In both groups ptiO2 increased significantly in the postoperative course, whereby ptiO2 in the TMLR group was significantly lower compared with the CABG group. CONCLUSION: Although the exact mechanism of action of TMLR remains unclear, ptiO2 and thus oxygen supply in the myocardium increased in patients undergoing TMLR at least in the early postoperative course. However, ptiO2 increased to a lesser extent compared with CABG. PMID- 14534069 TI - Does angiographic severity of coronary artery disease predict postoperative outcome after coronary artery bypass surgery? AB - OBJECTIVE: It is not clear whether the severity of coronary artery disease as assessed on angiography has an impact on the postoperative outcome after coronary artery bypass surgery (CABG). DESIGN: The angiographic status of 15 coronary arteries/segments of 2,233 patients who underwent isolated on-pump CABG was graded according to the following criteria: 1 = no stenosis; 2 = stenosis <50%; 3 = stenosis of 50-69%; 4 = stenosis of 70-89%; 5 = stenosis of 90-99%; 6 = vessel occlusion; and 7 = vessel is not visualized. RESULTS: Thirty-seven patients (1.7%) died during the in-hospital stay and 108 (4.8%) developed postoperatively low cardiac output syndrome. Multivariate analysis showed that along with other risk factors the overall coronary angiographic score was predictive of postoperative death (p = 0.03; OR: 1.027, 95% CI: 1.003-1.052) and of low cardiac output syndrome (p = 0.04; OR: 1.172, 95% CI: 1.010-1.218). The status of the proximal segment of the left circumflex coronary artery, the diagonal arteries and the left obtuse marginal arteries was most closely associated with adverse postoperative outcome. CONCLUSION: The angiographic status of coronary arteries has an impact on the immediate outcome after CABG. PMID- 14534070 TI - Unspecific elevation of plasma troponin-T and CK-MB after coronary surgery. AB - OBJECTIVE: Biochemical markers of myocardial injury are frequently elevated after cardiac surgery. It is generally accepted that release unrelated to permanent myocardial damage explains a proportion of these elevations. However, little is known about the magnitude and temporal characteristics of this diagnostic noise. One way to address this issue would be to study a group without permanent myocardial injury. DESIGN: The unique release kinetics of troponin-T (permanent myocardial injury causes a sustained release of structurally bound troponin) were used to identify patients with no or minimal permanent myocardial injury. Blood was sampled from patients undergoing coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB) before surgery, 3 and 8 h after unclamping the aorta, and each morning until postoperative day 4, for analysis of enzymes and troponin-T. From 302 consecutive patients a subgroup was identified that fulfilled the following criteria: (a) normalized troponin-T levels < or = postoperative day 4; (b) no ECG changes indicating myocardial injury. RESULTS: Seventy-seven patients fulfilled the criteria above and in this subgroup troponin T (2.08 +/- 1.42 microg/l; range 0.35-8.99 microg/l) peaked at the 3 h recording and creatine kinase monobasic (CK-MB) (28.6 +/- 11.3 microg/l; range 11.9-86.0 microg/l) peaked at the 8 h recording after unclamping the aorta. CONCLUSION: Substantial early elevations of plasma CK-MB and troponin-T occurred in patients with no or minimal permanent myocardial injury after CABG. Unspecific release was most pronounced during the timeframe that is usually studied to evaluate myocardial protective strategies or to compare revascularization procedures. PMID- 14534071 TI - The role of magnesium in the endothelial dysfunction caused by global ischemia followed by reperfusion: in vitro study of canine coronary arteries. AB - OBJECTIVE: To study the role of magnesium in the endothelial dysfunction of canine coronary arteries caused by cardiopulmonary bypass (CPB) global ischemia followed by reperfusion. DESIGN: Segments of canine coronary arteries were suspended in organ chambers to measure isometric contraction by prostaglandin F (2alpha), and relaxed by acetylcholine (ACh), sodium fluoride (NaF), calcium ionophore (A23187) and sodium nitroprusside (SNP) in crescent concentrations. The investigation protocol had groups with six dogs: CONTROL group (without CPB), CPB group (105 min of CPB without aortic cross-clamping), ISCH group (45 min of CPB with aortic cross-clamping), ISCH/REP group (45 min of aortic cross-clamping followed by 60 min of reperfusion). The coronary relaxations were evaluated with (phase I), without (phase II) and restored magnesium (phase III) to the organ bath. RESULTS: The presence of magnesium in the organ bath was associated with the greater relaxation in response to agonists of the nitric oxide production. The removal of magnesium from the organ bath was associated with the reduction in the intensity of vessel relaxation. The magnesium restoration to the organ bath was associated with the additional reduction in the intensity of relaxation with the exception of NaF that allowed re-acquisition of the relaxation observed in the presence of magnesium. CONCLUSION: This in vitro study demonstrates that magnesium ion favorably influences the nitric oxide production by the coronary endothelium, attenuating the endothelial dysfunction caused by global ischemia followed by reperfusion. PMID- 14534072 TI - Myocardial cooling for reperfusion injury protection achieved by organ specific hypothermic autologous perfusion. AB - OBJECTIVE: Whole body hypothermia has been suggested to reduce myocardial injury in patients with ST-segment elevation myocardial infarction. Because of the large human thermal mass, induction of generalized hypothermia is slow and the technique has encountered considerable side effects. The aim was to develop and validate a method for regional cooling during myocardial reperfusion using hypothermic autologous blood. DESIGN: In a myocardial ischemia-reperfusion pig model (n = 10), arterial blood was cooled in a closed circuit, and returned to the myocardium during reperfusion either through a perfusion catheter or through the guiding catheter. Myocardial temperatures were recorded using temperature electrodes. RESULTS: Stabile regional myocardial cooling was induced without complications within 4 min. Both flow rate and blood temperature had significant impact on temperature in the reperfused myocardium but did not influence systemic temperature. CONCLUSION: A method for organ specific hypothermic autologous arterial blood reperfusion has been developed and validated. The method is a simple and much faster alternative to systemic cooling and may have the potential to reduce myocardial injury in patients with acute myocardial infarction. PMID- 14534073 TI - Control of LH secretory-burst frequency and interpulse-interval regularity in women. AB - Hypothalamic neurons generate discrete bursts of gonadotropin-releasing hormone (GnRH) and thereby pulses of luteinizing hormone (LH) at randomly timed intervals centered on a probabilistic mean frequency. We tested the hypothesis that physiological mechanisms govern not only the number but also the stochastic dispersion of the GnRH/LH pulse-renewal process in humans; for example, in young women in the early (EF) and late (LF) follicular and midluteal (ML) phases of the menstrual cycle (n = 18) and in postmenopausal individuals (PM, n = 16). To this end, we quantify stochastic interpulse variability by way of the order independent, two-parameter Weibull renewal process (Keenan DM and Veldhuis J. Am J Physiol Regul Integr Comp Physiol 281: R1917-R1924, 2001) and the sequence specific, model-free approximate-entropy statistic (ApEn) (Pincus SM. Proc Natl Acad Sci USA 88: 2297-2301, 1991). Statistical testing unveiled 1) reduced probabilistic mean LH secretory-burst frequency (lower lambda of the Weibull distribution) in ML compared with each of EF, LF, and PM (P < 0.001); 2) quantifiably more regular LH interburst-interval sets (elevated gamma of the Weibull density) in PM than in each of EF, LF, and ML (P < 0.01); 3) uniquely prolonged latency to maximal LH secretion within individual secretory bursts in ML (P < 0.01); and 4) comparably mean random, sequential LH interburst-interval and mass values (normalized ApEn) among the distinct hormonal milieus. From these data, we postulate that sex steroids and age determine daily LH secretory-burst number, quantifiable pulse-renewal variability, and secretory-waveform evolution. PMID- 14534074 TI - Aerobic training improves exercise-induced lipolysis in SCAT and lipid utilization in overweight men. AB - The aim of this study was to investigate whether endurance training improves lipid mobilization and oxidation in overweight subjects. Eleven young men (25.6 +/- 1.4 yr and body mass index 27.7 +/- 0.2) performed a 4-mo training program consisting of practicing aerobic exercise 5 days/wk. Before and after the training period, lipid oxidation was explored during a 60-min exercise at 50% of peak O2 consumption by use of indirect calorimetry. Lipid mobilization and antilipolytic alpha2-adrenoceptor effect were also studied using the microdialysis method in abdominal subcutaneous adipose tissue (SCAT). After training, plasma nonesterified fatty acid (NEFA) levels, at rest and during exercise, were significantly lower than before (P < 0.001). Lipolysis in SCAT was significantly higher after than before training. An antilipolytic alpha2 adrenoceptor effect in SCAT was underlined during exercise before training and disappeared after. The respiratory exchange ratio was lower after training, i.e., the percentage of lipid oxidation was higher only at rest. The amount of lipid oxidized was higher after training, at rest, and during exercise. Although exercise power was higher after training, the relative intensity was equivalent, as suggested by a similar increase in plasma catecholamine concentrations before and after training. In conclusion, 4-mo training in overweight men improved lipid mobilization through a decrease of antilipolytic alpha2-adrenoceptor effect in SCAT and lipid oxidation during moderate exercise. Training induced a decrease of blood NEFA, predicting better prevention of obesity. PMID- 14534075 TI - Glucagon-like peptide-1 induces a cAMP-dependent increase of [Na+]i associated with insulin secretion in pancreatic beta-cells. AB - Glucagon-like peptide-1 (GLP-1) elevates the intracellular free calcium concentration ([Ca2+]i) and insulin secretion in a Na+-dependent manner. To investigate a possible role of Na ion in the action of GLP-1 on pancreatic islet cells, we measured the glucose-and GLP-1-induced intracellular Na+ concentration ([Na+]i), [Ca2+]i, and insulin secretion in hamster islet cells in various concentrations of Na+. The [Na+]i and [Ca2+]i were monitored in islet cells loaded with sodium-binding benzofuran isophthalate and fura 2, respectively. In the presence of 135 mM Na+ and 8 mM glucose, GLP-1 (10 nM) strongly increased the [Na+]i, [Ca2+]i, and insulin secretion. In the presence of 13.5 mM Na+, both glucose and GLP-1 increased neither the [Na+]i nor the [Ca2+]i. In a Na+-free medium, GLP-1 and glucose did not increase the [Na+]i. SQ-22536, an inhibitor of adenylate cyclase, and H-89, an inhibitor of PKA, incompletely inhibited the response. In the presence of both 8 mM glucose and H-89, 8-pCPT-2'-O-Me-cAMP, a PKA-independent cAMP analog, increased the insulin secretion and the [Na+]i. Therefore, we conclude that GLP-1 increases the cAMP level via activation of adenylate cyclase, which augments the membrane Na+ permeability through PKA dependent and PKA-independent mechanisms, thereby increasing the [Ca2+]i and promoting insulin secretion from hamster islet cells. PMID- 14534076 TI - Carbon monoxide stimulates insulin release and propagates Ca2+ signals between pancreatic beta-cells. AB - A key question for understanding the mechanisms of pulsatile insulin release is how the underlying beta-cell oscillations of the cytoplasmic Ca2+ concentration ([Ca2+]i) are synchronized within and among the islets in the pancreas. Nitric oxide has been proposed to coordinate the activity of the beta-cells by precipitating transients of [Ca2+]i. Comparing ob/ob mice and lean controls, we have now studied the action of carbon monoxide (CO), another neurotransmitter with stimulatory effects on cGMP production. A strong immunoreactivity for the CO producing constitutive heme oxygenase (HO-2) was found in ganglionic cells located in the periphery of the islets and in almost all islet endocrine cells. Islets from ob/ob mice had sixfold higher generation of CO (1 nmol.min-1.mg protein-1) than the lean controls. This is 100-fold the rate for their constitutive production of NO. Moreover, islets from ob/ob mice showed a threefold increase in HO-2 expression and expressed inducible HO (HO-1). The presence of an excessive islet production of CO in the ob/ob mouse had its counterpart in a pronounced suppression of the glucose-stimulated insulin release from islets exposed to the HO inhibitor Zn-protoporhyrin (10 microM) and in a 16 times higher frequency of [Ca2+]i transients in their beta-cells. Hemin (0.1 and 1.0 microM), the natural substrate for HO, promoted the appearance of [Ca2+]i transients, and 10 microM of the HO inhibitors Zn-protoporphyrin and Cr mesoporphyrin had a suppressive action both on the firing of transients and their synchronization. It is concluded that the increased islet production of CO contributes to the hyperinsulinemia in ob/ob mice. In addition to serving as a positive modulator of glucose-stimulated insulin release, CO acts as a messenger propagating Ca2+ signals with coordinating effects on the beta-cell rhythmicity. PMID- 14534077 TI - Insulin/IGF-I-signaling pathway: an evolutionarily conserved mechanism of longevity from yeast to humans. AB - Although the underlying mechanisms of longevity are not fully understood, it is known that mutation in genes that share similarities with those in humans involved in the insulin/insulin-like growth factor I (IGF-I) signal response pathway can significantly extend life span in diverse species, including yeast, worms, fruit flies, and rodents. Intriguingly, the long-lived mutants, ranging from yeast to mice, share some important phenotypic characteristics, including reduced insulin signaling, enhanced sensitivity to insulin, and reduced IGF-I plasma levels. Such genetic homologies and phenotypic similarities between insulin/IGF-I pathway mutants raise the possibility that the fundamental mechanism of aging may be evolutionarily conserved from yeast to mammals. Very recent findings also provide novel and intriguing evidence for the involvement of insulin and IGF-I in the control of aging and longevity in humans. In this study, we focus on how the insulin/IGF-I pathway controls yeast, nematode, fruit fly, and rodent life spans and how it is related to the aging process in humans to outline the prospect of a unifying mechanism in the genetics of longevity. PMID- 14534078 TI - Role of hypothalamic inputs in maintaining pituitary-adrenal responsiveness in repeated restraint. AB - The role of hypothalamic structures in the regulation of chronic stress responses was studied by lesioning the mediobasal hypothalamus or the paraventricular nucleus of hypothalamus (PVH). Rats were acutely (60 min) and/or repeatedly (for 7 days) restrained. In controls, a single restraint elevated the plasma adrenocorticotropin (ACTH), corticosterone, and prolactin levels. Repeated restraint produced all signs of chronic stress, including decreased body and thymus weights, increased adrenal weight, basal corticosterone levels, and proopiomelanocortin (POMC) mRNA expression in the anterior pituitary. Some adaptation to repeated restraint of the ACTH response, but not of other hormonal responses, was seen. Lesioning of the mediobasal hypothalamus abolished the hormonal response and POMC mRNA activation to acute and/or repeated restraint, suggesting that the hypothalamo-pituitary-adrenal axis activation during repeated restraint is centrally driven. PVH lesion inhibited the ACTH and corticosterone rise to the first restraint by approximately 50%. In repeatedly restrained rats with PVH lesion, the ACTH response to the last restraint was reduced almost to basal control levels, and the elevation of POMC mRNA level was prevented. PVH seems to be important for the repeated restraint-induced ACTH and POMC mRNA stimulation, but it appears to partially mediate other restraint-induced hormonal changes. PMID- 14534079 TI - Developmental switch in brain nutrient transporter expression in the rat. AB - Normal development of both human and rat brain is associated with a switch in metabolic fuel from a combination of glucose and ketone bodies in the immature brain to a nearly total reliance on glucose in the adult. The delivery of glucose, lactate, and ketone bodies from the blood to the brain requires specific transporter proteins, glucose and monocarboxylic acid transporter proteins (GLUTs and MCTs), respectively. Developmental expression of the GLUTs in rat brain, i.e., 55-kDa GLUT1 in the blood-brain barrier (BBB), 45-kDa GLUT1 and GLUT3 in vascular-free brain, corresponds to maturational increases in cerebral glucose uptake and utilization. It has been suggested that MCT expression peaks during suckling and sharply declines thereafter, although a comparable detailed study has not been done. This study investigated the temporal and regional expression of MCT1 and MCT2 mRNA and protein in the BBB and the nonvascular brain during postnatal development in the rat. The results confirmed maximal MCT1 mRNA and protein expression in the BBB during suckling and a decline with maturation, coincident with the switch to glucose as the predominant cerebral fuel. However, nonvascular MCT1 and MCT2 levels do not reflect changes in cerebral energy metabolism, suggesting a more complex regulation. Although MCT1 assumes a predominantly glial expression in postweanling brain, the concentration remains fairly constant, as does that of MCT2 in neurons. The maintenance of nonvascular MCT levels in the adult brain implies a major role for these proteins, in concert with the GLUTs in both neurons and astrocytes, to transfer glycolytic intermediates during cerebral energy metabolism. PMID- 14534080 TI - Model to assess muscle protein turnover: domain of validity using amino acyl-tRNA vs. surrogate measures of precursor pool. AB - Current models to measure protein turnover across muscle bed are based on many surrogate measures of amino acyl-tRNA. We measured muscle protein turnover based on tracer-to-tracee ratios of the stable isotopes of leucine, phenylalanine, and ketoisocaproate (KIC) in artery and vein and muscle amino acyl-tRNA and muscle tissue fluid (TF) in 26 healthy subjects. A three-compartment model calculation based on arteriovenous and tRNA measurements was first performed and its domain of validity assessed. The results were then compared with those using simpler approaches based on surrogate measures of tRNA such as those of TF and KIC and a one-compartment model based on arteriovenous amino acids. In 96% of cases, the model using tRNA was applicable, but only in a lower percentage of cases were the results using surrogate measures applicable. Protein breakdown, protein synthesis, and shunting of amino acids from artery to vein were consistently underestimated, and fluxes of amino acid from artery to intracellular compartment and from intracellular compartment to vein were overestimated, when surrogate measures were used. The one-compartment model also underestimated protein breakdown and synthesis. Measurements using tissue fluid gave results closer to those based on tRNA. In conclusion, a three-compartment model using arteriovenous samples and amino acyl-tRNA provides measurements of muscle protein turnover of acceptable precision in 96% of cases. The precision was unacceptable in a substantial percentage of cases, and the accuracy of the estimation of protein fluxes was significantly affected when surrogate measures were used. PMID- 14534081 TI - Stimulation of adhesion molecule expression in human endothelial cells (HUVEC) by adrenomedullin and corticotrophin. AB - Adrenomedullin (AM) and corticotrophin (ACTH) are both vasoactive peptides produced by a variety of cell types, including endothelial cells. Although AM and ACTH are considered to be important in the control of blood pressure and the response to stress, respectively, their role in inflammation and the immune response has not been clarified. This study shows, with the use of a cell-based ELISA, that AM and ACTH induce cell surface expression of the adhesion molecules E-selectin, VCAM-1, and ICAM-1 on human umbilical vein endothelial cells (HUVEC). Furthermore, this effect appears to be mediated in part via elevation of cAMP, given that both peptides elevate cAMP, the cell-permeable cAMP analog dibutyryl cAMP is able to mimic induction of all three cell adhesion molecules and the effect of AM and ACTH is inhibited by the adenylyl cyclase inhibitor SQ-22536. These findings demonstrate a role for AM and ACTH in the regulation of the immune and inflammatory response. PMID- 14534082 TI - Ca transients from Ca channel activity in rat cardiac myocytes reveal dynamics of dyad cleft and troponin C Ca binding. AB - The properties of the dyad cleft can in principle significantly impact excitation contraction coupling, but these properties are not easily amenable to experimental investigation. We simultaneously measured the time course of the rise in integrated Ca current (ICa) and the rise in concentration of fura 2 with Ca bound ([Ca-fura 2]) with high time resolution in rat myocytes for conditions under which Ca entry is only via L-type Ca channels and sarcoplasmic reticulum (SR) Ca release is blocked, and compared these measurements with predictions from a finite-element model of cellular Ca diffusion. We found that 1) the time course of the rise of [Ca-fura 2] follows the time course of integrated ICa plus a brief delay (1.36 +/- 0.43 ms, n = 6 cells); 2) from the model, high-affinity Ca binding sites in the dyad cleft at the level previously envisioned would result in a much greater delay (>/=3 ms) and are therefore unlikely to be present at that level; 3) including ATP in the model promoted Ca efflux from the dyad cleft by a factor of 1.57 when low-affinity cleft Ca binding sites were present; 4) the data could only be fit to the model if myofibrillar troponin C (TnC) Ca binding were low affinity (4.56 microM), like that of soluble troponin C, instead of the high-affinity value usually used (0.38 microM). In a "good model," the rate constants for Ca binding and dissociation were 0.375 times the values for soluble TnC; and 5) consequently, intracellular Ca buffering at the rise of the Ca transient is inferred to be low. PMID- 14534083 TI - Purification of interstitial cells of Cajal by fluorescence-activated cell sorting. AB - Interstitial cells of Cajal (ICC) in the gastrointestinal tract generate and propagate slow waves and mediate neuromuscular neurotransmission. Although damages to ICC have been described in several gastrointestinal motor disorders, analysis of their gene expression in health and disease has been problematic because of the difficulties in isolating these cells. Our goal was to develop techniques for large-scale purification of ICC. Murine ICC were identified in live gastrointestinal muscles with fluorescent Kit antibodies. Because this technique also labels resident macrophages nonspecifically, we attempted to separate ICC from these cells by fluorescence-activated cell sorting with or without immunomagnetic presorting. Efficacy and specificity of ICC purification were tested by quantitative RT-PCR of cell-specific markers. Fluorescence-based separation of small intestinal ICC from unlabeled cells and macrophages tagged with F4/80 antibodies yielded 30,000-40,000 cells and approximately 60-fold enrichment of c-kit mRNA. However, the macrophage marker CD68 was also enriched approximately 6-fold. Magnetic presorting of ICC did not significantly improve selectivity. After labeling contaminating cells with additional paramagnetic (anti-CD11b, -CD11c) and fluorescent antibodies (anti-CD11b) and depleting them by magnetic presorting, we harvested approximately 2,000-4,000 cells from single gastric corpus-antrum muscles and detected an approximately 30-fold increase in c kit mRNA, no enrichment of mast cells, and an approximately 4-fold reduction of CD68 expression. Adding labeled anti-CD45 antibody to our cocktail further increased c-kit enrichment and eliminated mast cells and macrophages. Smooth muscle cells and myenteric neurons were also depleted. We conclude that immunofluorescence-based sorting can yield ICC in sufficiently high numbers and purity to permit detailed molecular analyses. PMID- 14534086 TI - Can poetry make better doctors? Teaching the humanities and arts to medical students and residents at the University of California, Irvine, College of Medicine. AB - The Program in Medical Humanities & Arts at the University of California, Irvine, College of Medicine has been in existence for five years. The program was implemented to enhance aspects of professionalism including empathy, altruism, compassion, and caring toward patients, as well as to hone clinical communication and observational skills. It contains elective or required curriculum across all four years of medical school and required curriculum in two residency programs, organized according to structural principles of horizontal coherence, vertical complexity, and patient care applications. The program emphasizes small-group, interdisciplinary teaching and faculty development, and is notable for learners' use of creative projects to reflect on patients and themselves. Evaluation of the program indicates a positive response among learners. More systematic studies point to increases in empathy and positive attitudes toward the humanities as tools for professional development as a result of exposure to the program curriculum. Future directions include closer collaboration with the University of California, Irvine, Schools of the Arts and Humanities, involvement of local artists and writers, and development of a graduation with distinction in humanities for medical students. PMID- 14534084 TI - [Ca2+]i regulates trafficking of Cav1.3 (alpha1D Ca2+ channel) in insulin secreting cells. AB - Chronic exposure of pancreatic beta-cells to high concentrations of glucose impairs the insulin secretory response to further glucose stimulation. This phenomenon is referred to as glucose desensitization. It has been shown that glucose desensitization is associated with abnormal elevation of beta-cell basal intracellular free Ca2+ concentration ([Ca2+]i). We have investigated the relationship between the basal intracellular free Ca2+ and the L-type (Cav1.3) Ca2+ channel translocation in insulin-secreting cells. Glucose stimulation or membrane depolarization induced a nifedipine-sensitive Ca2+ influx, which was attenuated when the basal [Ca2+]i was elevated. Using voltage-clamp techniques, we found that changing [Ca2+]i could regulate the amplitude of the Ca2+ current. This effect was attenuated by drugs that interfere with the cytoskeleton. Immunofluorescent labeling of Cav1.3 showed an increase in the cytoplasmic distribution of the channels under high [Ca2+]i conditions by deconvolution microscopy. The [Ca2+]i-dependent translocation of Cav1.3 channel was also demonstrated by Western blot analysis of biotinylation/NeutrAvidin-bead-eluted surface proteins in cells preincubated at various [Ca2+]i. These results suggest that Cav1.3 channel trafficking is involved in glucose desensitization of pancreatic beta-cells. PMID- 14534087 TI - Humanities Education at Northwestern University's Feinberg School of Medicine. AB - The Medical Humanities and Bioethics Program at Northwestern University's Feinberg School of Medicine is responsible for humanities education in all four years of medical school: five units of the required four-year Patient, Physician, and Society course, 37 to 40 medical humanities seminars in years one and two, more than 125 ethics case conferences in third-year clerkships, and electives for fourth-year students. The program faculty also participate in ethics and humanities education in residencies, and the program offers an annual one-year fellowship. The program introduced the small-group teaching that now characterizes much of the school's curriculum, and its course units and seminars have been a resource for faculty development and curricular innovation. Drawing on literature, religion, ethics, philosophy of medicine, film, history, social and cultural anthropology, and jurisprudence, humanities education is designed to foster habits of discourse on social and moral issues in medicine. Small-group teaching and interactive learning are its central pedagogical methods. Essential to their successful use in a school that enrolls approximately 170 students each year is a large cadre of volunteer clinicians who serve as tutors and the college system, a four-part division of each class instituted by the 1993 curriculum reform. Students are evaluated on preparation, class participation, and regular writing assignments. All course units and seminars are pass/fail (as are all first- and second-year courses); tutors supply narrative comments. The courses themselves are thoroughly evaluated by students and reviewed both by the relevant faculty-student committee and at an annual curriculum retreat. PMID- 14534088 TI - Literature & medicine: humanities at the heart of health care: a hospital-based reading and discussion program developed by the maine humanities council. AB - Created by the Maine Humanities Council in 1997, Literature & Medicine: Humanities at the Heart of Health Care is a hospital-based humanities reading and discussion program for practicing health care professionals. To date, a total of 25 (65%) of Maine's hospitals have implemented the six-month program at least once, reaching over 900 participants. At the monthly meetings, participants discuss assigned readings-works of fiction, poetry, drama, and nonfiction that illuminate issues central to caring for people. Scholars selected to facilitate the discussion foster nonhierarchical, wide-ranging discussions of the texts. Participants connect the world of science with the world of lived experience and engage with humanistic perspectives and insights that help them do their work better. Literature & Medicine is unique in promoting statewide programs that involve a heterogeneous mix of veteran health care professionals. The program has been or will be implemented in seven other states, and organizers are exploring further expansion of the program through a national institute. PMID- 14534089 TI - Ethics, professionalism, and humanities at Michigan State University College of Human Medicine. AB - This article describes the variety of approaches used at Michigan State University's College of Human Medicine for teaching ethics, professionalism, and humanities to undergraduate medical students: courses in ethics and health policy; mentoring programs; selectives in history, literature, and spirituality; structured patient care experiences; and discussions with students in their clinical years on the ethical and professional challenges confronting them in their clinical experiences. Some of these approaches, such as the structured patient-care experience, may be unique to Michigan State. The authors place special emphasis on discussing the challenges that confront this curriculum, including struggles to keep up with the pace of change in the health care system, preserving and highlighting the linkages between the "ethics" and the "professionalism" strands of the curriculum, making optimal use of Web technologies, successfully communicating to students the ultimately practical importance of the medical humanities other than ethics, and solving the problems of geography created by a widely dispersed community campus system. PMID- 14534090 TI - The Program in Medical Humanities at the University of Missouri-Kansas City School of Medicine. AB - The University of Missouri-Kansas City School of Medicine opened officially in 1971, offering a combined BA/MD in a six-year program. The program has a defined goal of offering humanities courses throughout the curriculum. The Sirridge Office of Medical Humanities was established in 1992, and it assumed the responsibility of developing conveniently scheduled courses in medical humanities, making more specific requirements for such courses in the medical curriculum. The goal of these courses has been to provide students a different way of looking at things by giving them insight into the ways that reading literature and writing stories helps medical students better read patient experiences. A course entitled " LITERATURE: A Healing Art" was first offered in 1992. In 1994, The Body Image in Medicine and the Arts course was added. With the help of a Culpeper grant in 1995, five interdisciplinary undergraduate courses were created. In 1999 Medicine, LITERATURE, and Law was developed from two previous courses. Bioethics is offered as part of a series called Correlative Medicine and in a required course, Behavioral Science. The present director of the Office of Medical Humanities has resigned and at present there is a search for a replacement. A new director may decide to change some of the offerings and add new activities. The medical school remains committed to supporting the office and its programs in keeping with the goals of the original academic plan to educate safe and caring physicians. PMID- 14534091 TI - Medical humanities at New York University School of Medicine: an array of rich programs in diverse settings. AB - The New York University School of Medicine has a rich tradition of cultivating programs in medical humanities and professionalism. They are drawn from the departments, centers, students, and faculty in the School of Medicine, have linkages throughout the university, and are interwoven into the fabric and culture of the institution. Some are centrally based in the School of Medicine's deans' office, and others are located in individual departments and receive support from the dean's office. This article describes representative programs for medical students and faculty. Curricular initiatives, the fundamental components of medical students' learning, include a course entitled "The Physician, Patient, and Society," a clerkship essay in the Medicine Clerkship, an opportunity for reflection during the medicine clerkship, and a medical humanities elective. In 2002, the Professionalism Initiative was launched to enhance and reflect the values of the medical profession. Its curriculum consists of a series of events that coordinate, particularly, with existing elements of the first-year curriculum (e.g., orientation week, a session during anatomy, a self-assessment workshop, and a peer-assessment workshop). The Master Scholars Program is a group of five, theme-based master societies consisting of faculty and students who share common interests around the society's themes. Programs developed for the societies include colloquia, faculty-led seminars, a mandatory student-mentoring program, and visiting scholars. Finally, the authors describe three high-quality literary publications created at New York University School of Medicine. Each of the initiatives undergoes regular critical examination and reflection that drive future planning. PMID- 14534093 TI - Developing a medical humanities concentration in the medical curriculum at the University of Rochester School of Medicine and Dentistry, Rochester, New York, USA. AB - To the author's knowledge, the University of Rochester School of Medicine and Dentistry is the only medical school in the United States that offers a concentration or minor in medical humanities for medical students. This article presents how the author first thought of offering a concentration in medical humanities and explains the educational elements students must satisfy. In 1998, the university underwent a major curriculum revision, dubbed the "double-helix" curriculum because of its goal of intertwining basic science and clinical medicine over all four years of medical school. As course director of the Medical Humanities Seminars for more than ten years, the author saw this change as an opportunity to expand the humanities curriculum. The number of sessions and courses offered in the first two years doubled as part of the transition to the new curriculum. In addition, the author proposed to the medical school curriculum steering committee to approve a concentration in clinical ethics and humanities. The concentration option motivates students to continue to pursue their humanistic interests in the third and fourth years of medical school. About 25% of the student body has signed up in the first two years the concentration has been available. PMID- 14534094 TI - Human contexts: Medicine in Society at Stony Brook University School of Medicine. AB - Humanities teaching was introduced at Stony Brook University School of Medicine by Edmund Pellegrino, the first dean of the Medical School and founder of the Health Sciences Center. Since 1990, "Medicine in Society" has been a substantial presence throughout the curriculum, introducing students to the perspectives of a wide range of humanities disciplines as they apply to health care, and continuing as a sustained presence throughout the four years of training. Medicine in Society serves as a reminder that medicine is a human and communal endeavor, situated in sociocultural contexts, reliant on human values, and articulated most often through narratives. The authors describe the structure and function of the Medicine in Society curriculum and the Institute for Medicine in Contemporary Society, summarize their evaluation of the program, and outline their plans for meeting current and future challenges. PMID- 14534095 TI - The Biomedical Humanities program: merging humanities and science in a premedical curriculum at Hiram College. AB - The Biomedical Humanities program at Hiram College, established in 1999, engages premedical and other qualified students in ethical and informed decision making, improves their ability to interact with persons of different backgrounds and cultures, provides them an active introduction to basic medical research and clinical practice, and coaches them in communicating across barriers, appreciating that scientists and humanists typically learn and work differently. The program offers both a major and a minor in biomedical humanities topics. The major requires the core biology and chemistry curriculum necessary for further studies in medicine as well as courses in genetics and statistics. The remainder of the major is devoted to four core areas: Communications, Relationships and Cultural Sensitivity, Ethics and Medical Humanities, and a nonacademic core area, Experiential Learning. Many of the ethics and medical humanities options are team taught interdisciplinary courses. The Experiential Learning area requires students to take two special topics seminars, two service seminars, and two internships-one shadowing a professional in his or her area of interest and one engaging in basic biomedical research. The shadowing internship and service seminars focus not only on career exploration, but also on human interactions. Students reflect on the personal interactions they observe during their various experiences, and on their own strengths and weaknesses. Essays, designed to help students learn more about their roles in these settings, push them to deal with the sociopolitical issues involved in their service. The major, a robust and vital component of Hiram's undergraduate program, has attracted academically gifted students with a diverse array of career goals. PMID- 14534096 TI - The Medical Humanities at the Northeastern Ohio Universities College of Medicine: historical, theoretical, and curricular perspectives. AB - The Human Values in Medicine Program (HVM) at the Northeastern Ohio Universities College of Medicine is composed of 120 required hours in medical humanities, social sciences, and behavioral sciences. In addition to a required HVM month in the fourth year when the bulk of the course work is completed, students can also choose from lectures, seminars, and short courses during the first, second, and third years. The broad goal of the HVM program as originally conceived was for students to use the content and skills of the humanities disciplines to reflect on their own and others' values, and to appraise their role in the patient physician relationship, both in the community and in the larger culture. During the past several years, a cultural studies orientation has also been included, particularly the practice of critical analysis aimed at identifying the inequities and injustices within the doctor-patient relationship, in medical training, and in health care access and delivery in the U.S. and beyond. Current program development includes standardizing a bioethics curriculum for all students and developing a required fourth-year course that all students take during their HVM elective month. PMID- 14534097 TI - Humanities education at Pennsylvania State University College of Medicine, Hershey, Pennsylvania. AB - The Department of Humanities at Pennsylvania State University's (Penn State) College of Medicine, created at the founding of the College in 1967, was the first of its kind at any medical school. This article begins by describing how the department has developed over the years, and then discusses its present configuration, including kinds of faculty appointments, information about how it is funded, specific courses that comprise the department's four-year curriculum, and activities it sponsors. That a College of Medicine would make the teaching and practice of humanistic medicine a major and explicit commitment attracted the notice of Drs. Lawrence and Jane Kienle, who made possible the Doctors Kienle Center for Humanistic Medicine. The Humanities Department is closely allied with the Doctors Kienle Center, which integrates the College and the Medical Center by supporting a variety of projects, research, and awards that further the teaching and practice of humanistic medicine. Faculty in the department are encouraged to demonstrate excellence in scholarship and in teaching, but are also expected to become engaged in the life of the Medical Center, especially in furthering humanistic patient care. As such, the Humanities Department plays a major role in furthering the mission of the Pennsylvania State University's College of Medicine. PMID- 14534098 TI - Medical Humanities at the University of Texas Medical Branch at Galveston. AB - The Institute for the Medical Humanities of the University of Texas Medical Branch at Galveston (UTMB) was established in June 1973 to ensure that humanities teaching and research became an integral part of the education of future scientists and health-care professionals at UTMB. The multidisciplinary faculty of the Institute-who currently represent the disciplines of art, drama, history, law, literature, philosophy, and religious studies-teach in all four years of the undergraduate medical curriculum as well as in various residency programs. In addition to its focus on students and residents in the School of Medicine, the Institute has a vibrant graduate program in medical humanities with several joint degree options, including an MD/MA and an MD/PhD program, and the Institute has always included the School of Nursing, the School of Allied Health Sciences, and the Graduate School of Biomedical Sciences in its activities. After 30 years, the Institute's commitment to health-professional education remains strong and enduring. Challenged by major curriculum reform in the School of Medicine and increasingly tight state budgets, Institute faculty members look forward to continued collaboration with their basic science and clinical colleagues to improve our evolving curriculum and to seek research funding from external sources. PMID- 14534099 TI - On Becoming a Humanities Curriculum: the Center for Medical Humanities and Ethics at the University of Texas Health Science Center at San Antonio. AB - The authors describe the development and implementation of a new longitudinal and integrated humanities curriculum in the School of Medicine at the University of Texas Health Science Center at San Antonio. Included are a history of ethics and humanities education in the medical school leading to the creation of the Center for Medical Humanities and Ethics in July 2002; an articulation of the Center's basic principles; a curricular blueprint outlining the core objectives and methodologies, such as a narrative approach of this required, four-year humanities curriculum for undergraduate medical students; and an overview of course materials and activities. PMID- 14534100 TI - The Medical Humanities Program at the University of Manitoba, Winnipeg, Manitoba, Canada. AB - The current Medical Humanities Program at the University of Manitoba has evolved from a series of voluntary sessions into an integral element of the curriculum since its inception as the Human Values Program in 1986. With strong academic and financial support, the Medical Humanities Program has greatly benefited from dedicated leadership and a commitment to ongoing curricular review and redevelopment. The current Medical Humanities Program comprises six distinct components: Clinical Ethics; History of Medicine; Law; Complementary and Alternative Medicine; Palliative Care; and Human Values. Each of these components is compulsory and the first five are tested through examinations and assignments. Human Values sessions are designed to be experiential and to explore the human side of medicine as well as the intersections between medicine and the arts, literature, social psychology, and spirituality. The authors outline the origins and evolution of this successful program and describe its current components, student and faculty opinions, funding, advantages, disadvantages, and anticipated growth. PMID- 14534101 TI - Development of a Medical Humanities Program at Dalhousie University Faculty of Medicine, Nova Scotia, Canada, 1992-2003. AB - The Medical Humanities Program at Dalhousie University Faculty of Medicine in Nova Scotia, Canada, was initiated in 1992 to incorporate the medical humanities into the learning and experiences of medical students. The goal of the program was to gain acceptance as an integral part of the medical school. The program assumed a broad concept of the medical humanities that includes medical history, literature, music, art, multiculturalism, philosophy, epistemology, theology, anthropology, professionalism, history of alternative therapies, writing, storytelling, health law, international medicine, and ethics. Phase I of the program has provided the same elective and research opportunities in the medical humanities that are available to the students in clinical and basic sciences, and has encouraged and legitimized the involvement of the humanities in the life and learning of the medical student through a wide array of programs and activities. Phase II will focus on further incorporation of the humanities into the curriculum. Phase III will be the development of a graduate program in medical humanities to train more faculty who will incorporate the humanities into their teaching and into the development of education programs. PMID- 14534102 TI - Teaching Humanities at the National University of la Plata, Argentina. AB - In this article the author begins by giving a brief history of medical education in Argentina, then provides some background information on the National University of La Plata. He describes two major initiatives at La Plata: a new and pioneering admission policy (implemented in 1993) and a change in the number of hours and years in the curriculum. He then looks back to the introduction in 1976 of La Plata's medical humanities program. Over its 20 year existence, the program has undergone a number of changes in response both to students' interests and financial concerns. The revised humanities curriculum now consists of four elective courses: medicine and literature, anthropology, history of medicine, and "medical kalology" (which focuses on music and dance). Unfortunately, the program, while it has been well received by both students and faculty and has inspired programs at other Argentinean schools, is threatened by Argentina's economic crisis. PMID- 14534103 TI - The history of medicine teaching program in Croatia. AB - A history of medicine program was first introduced at the University of Zagreb, Croatia, in 1927 by the internist Lujo Thaller, who became the first lecturer on this subject at Zagreb. However, the development of the program did not go smoothly and medicohistorians in Croatia were engaged in a constant struggle for proper status and a permanent place for their program within the medical school curriculum. Today a 15-hour medical history course (offered in the last semester of the 6-year curriculum) is mandatory at all four Croatian medical schools. The course draws on historical, cultural, sociological, epistemological, and bioethical aspects of medicine, with the goal of broadening students' perspectives on the practice of medicine and thereby enabling them to better meet the demands they will face as practicing physicians. PMID- 14534104 TI - A medical humanities special study module on principles of medical theory and practice at the Charite, Humboldt University, Berlin, Germany. AB - The authors are members of a committee in charge of a special study module (SSM) entitled Principles of Medical Theory and Practice in a problem-based and integrated reformed curriculum track at the Charite, the medical school and university hospital of the Humboldt University, Berlin, Germany. The SSM contextualizes medicine by highlighting the societal contexts of the doctor patient relationship and the medical profession. Integrating the humanities into medical education helps students develop an awareness of the strengths and limitations of modern medicine, develop their own personalities and sense of social responsibility, and generally broaden their outlook. Teachers in the SSM seminars are from different disciplines, such as the history of medicine, bioethics, sociology, anthropology, and complementary medicine. Once a week, one or two teachers meet with as many as 21 students per group for a 90-minute course. Twelve courses constitute a seminar. Students are required to participate in four seminars during five years of studies. They can choose different topics from a set range. Although this SSM has been largely successful, some problems have occurred. Results from the course evaluations and experiences show that the seminars differ from one another in many ways. Financial restraints and the departmental structure of the faculty have influenced implementation of the SSM. However, the SSM is a new concept and is continuously reviewed and renewed. Future plans will be to specify outcomes, continue to discuss reasonable seminar topics, establish continuous support and training for teachers, and motivate students to become actively involved in the seminar discussions. PMID- 14534105 TI - Medicine and the arts in the undergraduate medical curriculum at the University of Oslo Faculty of Medicine, Oslo, Norway. AB - The authors describe a course titled Medicine and the Arts, established in 1996, that is part of the first semester of the undergraduate medical curriculum at the University of Oslo Faculty of Medicine. The course comprises four two-hour seminars on literature, visual arts, architecture, and music. The core objectives of the course are to demonstrate how art can be a source of personal and professional development, and also how art represents a source of insight into patients' experiences and the social, cultural, and historical context of medical practice. The course emphasizes that art is ambiguous and many layered and that its interpretation requires sensitivity, engagement, imagination, and reflection. Fostering these skills is a major aim of the course because these skills are also essential for clinical competence and professional development. The course's seminars are integrated into the curriculum but are not compulsory. Although the topics covered by the course have not been explicitly addressed in the formal examination of students, there has been some discussion about doing so, which would be a signal that those topics are as important as others in the curriculum. PMID- 14534106 TI - The Humanistic Medicine program at the Karolinska Institute, Stockholm, Sweden. AB - In 1998, the Humanistic Medicine program was established at the Karolinska Institute, Stockholm, Sweden. A fundamental element of the program is to promote medical humanities within clinical practice. The program's design focuses on three interconnected areas of study, the history of medicine, philosophy of medical science and practice, and aspects of the clinical encounter. The program offers undergraduate and postgraduate studies. The program's humanities content is bolstered in the medical curriculum by The Doctor School, a line of teaching medical students follow through their first four semesters. From this parallel series of lectures and seminars, students are exposed to further humanities and medical training. Students also have the option to select from humanities courses for their 17 eligible weeks of electives. It is hoped that the Karolinska Institute will continue to develop the humanities content of its curriculum, intertwining scientific exploration and humanistic understanding. PMID- 14534107 TI - Medical humanities: a new undergraduate teaching program at the University of Geneva School of Medicine, Switzerland. AB - In 2001, a new program of medical humanities was initiated at the University of Geneva School of Medicine in Switzerland. Four mandatory seminars and one optional 2-week internship are offered to second-through fifth-year medical students. The program has four interdependent goals: contextualizing, developing personal reflection and judgment, encouraging imagination, and offering specific ways to improve the quality of the therapeutic relationship. The program is based on an integrated vision of the humanities and stimulates the students' imagination and reflection in a way that medical students should find useful. Three steps help teachers to build an integrated vision: familiarization, confrontation, and adjustment to the medical culture. The mandatory seminars are taught by a team consisting of a physician and a humanities teacher. All the physicians, department heads, and clinicians involved in each seminar actively collaborated. The medical humanities program is in the Bioethics Unit, which is housed in the Department of Community Health and Medicine with medical history and legal medicine. This intellectual, institutional, and physical proximity encourages informal dialogue and ensures a more coherent and unified vision of the different disciplines. In their assessments of the program, students stated that the seminars gave them food for thought and met their expectations. However, it is premature to draw conclusions from these assessments because the program is still in its infancy. The program strives to provide students with tools specific to the humanities so that they can strengthen their own judgment, listening skills, open-mindedness, creativity, and curiosity, attributes that are needed to ensure that the therapeutic relationship will be satisfying for both physicians and patients. PMID- 14534108 TI - The Centre for Medical Humanities, Royal Free and University College Medical School, London, England. AB - The Centre for Medical Humanities is based within the Royal Free and University College Medical School. With a student body of more than 2,000, this large United Kingdom medical school benefits from being a fully integrated component of University College London. With its strong humanities and science faculties, University College London was founded more than 175 years ago to make education available to wider social and religious groups. With this background, it is logistically and ideologically well suited to sustaining a medical humanities program. The Centre's aims are to raise awareness of the field of medical humanities, to develop resources for the academic and teaching communities, and to build a broad and well-grounded educational program supported by sound educational principles and innovative research. Since 1998, the Centre and its predecessor, the Medical Humanities Unit, which was established by a group of medical educators with a diverse range of interests in the arts and humanities, has built interdisciplinary links and collaborative relationships with individuals and institutions within the university and beyond, nationally and internationally. These links and relationships have provided invaluable contributions and stimulus to the Centre's activities. The undergraduate educational program described in this article includes core and optional teaching throughout the curriculum, and a 1-year intercalated Bachelor of Science in Medical Humanities degree. For postgraduates, the Centre offers the United Kingdom's first continuing professional development accredited 2-day course in medical humanities, and an annual residential retreat open only to graduates of the course. PMID- 14534109 TI - Annotation of Toni Morrison's Recitatif. PMID- 14534110 TI - Commentary. Reference system for the fiction and poetry. PMID- 14534111 TI - A humanistic role model in my medical career. PMID- 14534113 TI - University of Arkansas College of Medicine, division of medical humanities. PMID- 14534114 TI - Stanford University School of Medicine, Arts and Humanities Medical Scholars Program. PMID- 14534115 TI - University of California, San Francisco, School of Medicine, Department of Anthropology, History and Social Medicine. PMID- 14534116 TI - University of South Florida, Program in Bioethics and Medical Humanities. PMID- 14534117 TI - Southern Illinois University School of Medicine, Department of Medical Humanities. PMID- 14534118 TI - University of Illinois at Chicago, Medical Humanities Program. PMID- 14534119 TI - University of Michigan Medical School, Program in Society and Medicine. PMID- 14534120 TI - Saint Patrick Hospital and Health Sciences Center and the University of Montana, Institute of Medicine and Humanities. PMID- 14534121 TI - Davidson College, Medical Humanities Program. PMID- 14534122 TI - University of Nebraska College of Medicine, section on humanities and law, Department of Preventive and Societal Medicine. PMID- 14534123 TI - The Brody School of Medicine at East Carolina University, Department of Medical Humanities. PMID- 14534124 TI - Drexel University College of Medicine, Medicine and the Arts online. PMID- 14534125 TI - University of Virginia School of Medicine, Program of Humanities in Medicine. PMID- 14534126 TI - Memorial University of Newfoundland Faculty of Medicine, the Medical Humanities in the clinical skills course. PMID- 14534127 TI - University of Sydney, Medical Humanities Program. PMID- 14534128 TI - University Hospital Hamburg-Eppendorf, University of Hamburg, Institute for the History and Ethics of Medicine. PMID- 14534129 TI - Faculty of Medicine of the Technion-Israel Institute of Technology, Humanities in Medicine. PMID- 14534130 TI - University of Auckland, Faculty of Medical and Health Sciences, Medical Humanities courses. PMID- 14534131 TI - National Taiwan University College of Medicine, the design of medical humanities courses for clerkships. PMID- 14534132 TI - University of Durham, Centre for Arts and Humanities in Health and Medicine. PMID- 14534133 TI - lockjaw encodes a zebrafish tfap2a required for early neural crest development. AB - The neural crest is a uniquely vertebrate cell type that gives rise to much of the craniofacial skeleton, pigment cells and peripheral nervous system, yet its specification and diversification during embryogenesis are poorly understood. Zebrafish homozygous for the lockjaw (low) mutation show defects in all of these derivatives and we show that low (allelic with montblanc) encodes a zebrafish tfap2a, one of a small family of transcription factors implicated in epidermal and neural crest development. A point mutation in low truncates the DNA binding and dimerization domains of tfap2a, causing a loss of function. Consistent with this, injection of antisense morpholino oligonucleotides directed against splice sites in tfap2a into wild-type embryos produces a phenotype identical to low. Analysis of early ectodermal markers revealed that neural crest specification and migration are disrupted in low mutant embryos. TUNEL labeling of dying cells in mutants revealed a transient period of apoptosis in crest cells prior to and during their migration. In the cranial neural crest, gene expression in the mandibular arch is unaffected in low mutants, in contrast to the hyoid arch, which shows severe reductions in dlx2 and hoxa2 expression. Mosaic analysis, using cell transplantation, demonstrated that neural crest defects in low are cell autonomous and secondarily cause disruptions in surrounding mesoderm. These studies demonstrate that low is required for early steps in neural crest development and suggest that tfap2a is essential for the survival of a subset of neural crest derivatives. PMID- 14534134 TI - AXR3 and SHY2 interact to regulate root hair development. AB - Signal transduction of the plant hormone auxin centres on the regulation of the abundance of members of the Aux/IAA family of transcriptional regulators, of which there are 29 in Arabidopsis. Auxin can influence Aux/IAA abundance by promoting the transcription of Aux/IAA genes and by reducing the half-life of Aux/IAA proteins. Stabilising mutations, which render Aux/IAA proteins resistant to auxin-mediated degradation, confer a wide range of phenotypes consistent with disruptions in auxin response. Interestingly, similar mutations in different family members can confer opposite phenotypic effects. To understand the molecular basis for this functional specificity in the Aux/IAA family, we have studied a pair of Aux/IAAs, which have contrasting roles in root hair development. We have found that stabilising mutations in AXR3/IAA17 blocks root hair initiation and elongation, whereas similar mutations in SHY2/IAA3 result in early initiation of root hair development and prolonged hair elongation, giving longer root hairs. The phenotypes resulting from double mutant combinations, the transient induction of expression of the proteins, and the pattern of transcription of the cognate genes suggest that root hair initiation is controlled by the relative abundance of SHY2 and AXR3 in a cell. These results suggest a general model for auxin signalling in which the modulation of the relative abundance of different Aux/IAA proteins can determine which down-stream responses are induced. PMID- 14534135 TI - LET-99 opposes Galpha/GPR signaling to generate asymmetry for spindle positioning in response to PAR and MES-1/SRC-1 signaling. AB - G-protein signaling plays important roles in asymmetric cell division. In C. elegans embryos, homologs of receptor-independent G protein activators, GPR-1 and GPR-2 (GPR-1/2), function together with Galpha (GOA-1 and GPA-16) to generate asymmetric spindle pole elongation during divisions in the P lineage. Although Galpha is uniformly localized at the cell cortex, the cortical localization of GPR-1/2 is asymmetric in dividing P cells. In this report, we show that the asymmetry of GPR-1/2 localization depends on PAR-3 and its downstream intermediate LET-99. Furthermore, in addition to its involvement in spindle elongation, Galpha is required for the intrinsically programmed nuclear rotation event that orients the spindle in the one-cell. LET-99 functions antagonistically to the Galpha/GPR-1/2 signaling pathway, providing an explanation for how Galpha dependent force is regulated asymmetrically by PAR polarity cues during both nuclear rotation and anaphase spindle elongation. In addition, Galpha and LET-99 are required for spindle orientation during the extrinsically polarized division of EMS cells. In this cell, both GPR-1/2 and LET-99 are asymmetrically localized in response to the MES-1/SRC-1 signaling pathway. Their localization patterns at the EMS/P2 cell boundary are complementary, suggesting that LET-99 and Galpha/GPR 1/2 signaling function in opposite ways during this cell division as well. These results provide insight into how polarity cues are transmitted into specific spindle positions in both extrinsic and intrinsic pathways of asymmetric cell division. PMID- 14534136 TI - C. elegans ankyrin repeat protein VAB-19 is a component of epidermal attachment structures and is essential for epidermal morphogenesis. AB - Elongation of the epidermis of the nematode Caenorhabditis elegans involves both actomyosin-mediated changes in lateral epidermal cell shape and body muscle attachment to dorsal and ventral epidermal cells via intermediate filament/hemidesmosome structures. vab-19 mutants are defective in epidermal elongation and muscle attachment to the epidermis. VAB-19 is a member of a conserved family of ankyrin repeat-containing proteins that includes the human tumor suppressor Kank. In epidermal cells, VAB-19::GFP localizes with components of epidermal attachment structures. In vab-19 mutants, epidermal attachment structures form normally but do not remain localized to muscle-adjacent regions of the epidermis. VAB-19 localization requires function of the transmembrane attachment structure component Myotactin. vab-19 mutants also display aberrant actin organization in the epidermis. Loss of function in the spectrin SMA-1 partly bypasses the requirement for VAB-19 in elongation, suggesting that VAB-19 and SMA-1/spectrin might play antagonistic roles in regulation of the actin cytoskeleton. PMID- 14534137 TI - Stepwise formation of a SMAD activity gradient during dorsal-ventral patterning of the Drosophila embryo. AB - Genetic evidence suggests that the Drosophila ectoderm is patterned by a spatial gradient of bone morphogenetic protein (BMP). Here we compare patterns of two related cellular responses, both signal-dependent phosphorylation of the BMP regulated R-SMAD, MAD, and signal-dependent changes in levels and sub-cellular distribution of the co-SMAD Medea. Our data demonstrate that nuclear accumulation of the co-SMAD Medea requires a BMP signal during blastoderm and gastrula stages. During this period, nuclear co-SMAD responses occur in three distinct patterns. At the end of blastoderm, a broad dorsal domain of weak SMAD response is detected. During early gastrulation, this domain narrows to a thin stripe of strong SMAD response at the dorsal midline. SMAD response levels continue to rise in the dorsal midline region during gastrulation, and flanking plateaus of weak responses are detected in dorsolateral cells. Thus, the thresholds for gene expression responses are implicit in the levels of SMAD responses during gastrulation. Both BMP ligands, DPP and Screw, are required for nuclear co-SMAD responses during these stages. The BMP antagonist Short gastrulation (SOG) is required to elevate peak responses at the dorsal midline as well as to depress responses in dorsolateral cells. The midline SMAD response gradient can form in embryos with reduced dpp gene dosage, but the peak level is reduced. These data support a model in which weak BMP activity during blastoderm defines the boundary between ventral neurogenic ectoderm and dorsal ectoderm. Subsequently, BMP activity creates a step gradient of SMAD responses that patterns the amnioserosa and dorsomedial ectoderm. PMID- 14534138 TI - Nkx6.1 controls migration and axon pathfinding of cranial branchio-motoneurons. AB - As many studies have focused on the mechanisms of motoneuron specification, little is known about the factors that control the subsequent development of postmitotic motoneurons. Previously, we showed that the transcription factor Nkx6.1 is required for the early specification of somatic motoneuron progenitors in the spinal cord. Our present analysis of hindbrain motoneuron development in Nkx6.1-deficient mouse embryos reveals that the early specification of branchio motoneurons is independent of Nkx6.1 function, but that it is required for their subsequent development. In Nkx6.1 mutant mice, we observed defects in the migration, as well as in the axon projections of branchio-motoneurons. A detailed analysis of the migratory defect in facial branchio-motoneurons reveals ectopic expression of the cell surface receptors Ret and Unc5h3 in premigratory neurons, but no changes in the rhombomeric environment. Taken together, our findings demonstrate a requirement for Nkx6.1 in the development of postmitotic motoneurons, and suggest a cell-autonomous function in the control of branchio motoneuron migration. PMID- 14534139 TI - Noradrenergic neurons in the zebrafish hindbrain are induced by retinoic acid and require tfap2a for expression of the neurotransmitter phenotype. AB - Tfap2a is a transcriptional activator expressed in many different cell types, including neurons, neural crest derivatives and epidermis. We show that mutations at the zebrafish locus previously called mont blanc (mob) or lockjaw (low) encode tfap2a. The mutant phenotype reveals that tfap2a is essential for the development of hindbrain noradrenergic (NA) neurons of the locus coeruleus, medulla and area postrema, as well as for sympathetic NA neurons, epibranchial placode derived visceral sensory ganglia, and craniofacial and trunk crest derivatives. We focus our analysis on the role of tfap2a NA differentiation in the CNS. In the locus coeruleus, Phox2a and Tfap2a are co-expressed and are both required for NA development. By contrast, in the medulla Phox2a and Tfap2a are expressed in adjacent overlapping domains, but only tfap2a activity is required for NA differentiation, as NA neurons develop normally in soulless/phox2a mutant medulla. phox2a and tfap2a do not appear to affect each others expression. Our studies show that two distinct inductive mechanisms control NA development in the zebrafish hindbrain. For the posterior hindbrain, we identify retinoic acid as an important signal to induce NA differentiation in the medulla oblongata and area postrema, where it expands the tfap2a expression domain and thus acts upstream of tfap2a. By contrast, previous work revealed Fgf8 to be involved in specification of NA neurons in the locus coeruleus. Thus, although the inductive signals may be distinct, hindbrain NA neurons of the locus coeruleus and the posterior groups both require Tfap2a to establish their noradrenergic identity. PMID- 14534140 TI - Caspase-independent cell engulfment mirrors cell death pattern in Drosophila embryos. AB - Programmed cell death plays an essential role during Drosophila embryonic development. A stereotypic series of cellular changes occur during apoptosis, most of which are initiated by a caspase cascade that is triggered by a trio of proteins, RPR, HID and GRIM. The final step in apoptosis is engulfment of the cell corpse. To monitor cell engulfment in vivo, we developed a fluorogenic beta galactosidase substrate that is cleaved by an endogenous, lysosomal beta galactosidase activity. The pattern of cell engulfment in wild-type embryos correlated well with the known pattern of apoptosis. Surprisingly, the pattern of cell engulfment persisted in apoptosis-deficient embryos. We provide evidence for a caspase-independent engulfment process that affects the majority of cells expected to die in developing Drosophila embryos. PMID- 14534141 TI - Multimodal tangential migration of neocortical GABAergic neurons independent of GPI-anchored proteins. AB - Neuronal migration is crucial for the construction of neuronal architecture such as layers and nuclei. Most inhibitory interneurons in the neocortex derive from the basal forebrain and migrate tangentially; however, little is known about the mode of migration of these neurons in the cortex. We used glutamate decarboxylase (Gad)67-green fluorescent protein (GFP) knock-in embryonic mice with expression of GFP in gamma-aminobutyric acid (GABA)-ergic neurons and performed time-lapse analysis. In coronal slices, many GFP-positive neurons in the lower intermediate zone (IZ) and subventricular zone (SVZ) showed robust tangential migration from lateral to medial cortex, while others showed radial and non-radial migration mostly towards the pial surface. In flat-mount preparations, GFP-positive neurons of the marginal zone (MZ) showed multidirectional tangential migration. Some of these neurons descended toward the cortical plate (CP). Intracortical migration of these neurons was largely unaffected by a treatment that cleaves glycosylphosphatidylinositol (GPI) anchors. These findings suggest that tangential migration of cortical interneurons from lateral to medial cortex predominantly occurs in the IZ/SVZ and raise the possibility that a part of the pial surface-directed neurons in the IZ/SVZ reach the MZ, whereby they spread into the whole area of the cortex. At least a part of these neurons may descend toward the CP. Our results also suggest that intracortical migration of GABAergic neurons occurs independent of GPI-anchored proteins. PMID- 14534142 TI - Embryonic handedness choice in C. elegans involves the Galpha protein GPA-16. AB - The mechanism by which polarity of the left-right (LR) axis is initially established with the correct handedness is not understood for any embryo. C. elegans embryos exhibit LR asymmetry with an invariant handedness that is first apparent at the six-cell stage and persists throughout development. We show here that a strong loss-of-function mutation in a gene originally designated spn-1 affects early spindle orientations and results in near randomization of handedness choice. This mutation interacts genetically with mutations in three par genes that encode localized cortical components. We show that the spn-1 gene encodes the Galpha protein GPA-16, which appears to be required for centrosomal association of a Gbeta protein. We will henceforth refer to this gene as gpa-16. These results demonstrate for the first time involvement of heterotrimeric G proteins in establishment of embryonic LR asymmetry and suggest how they might act. PMID- 14534144 TI - Emerging concept in angiogenesis: specification of arterial and venous endothelial cells. PMID- 14534143 TI - Fishing for novel angiogenic therapies. AB - The zebrafish has recently emerged as an important model for the study of vascular embryogenesis. Its genetic accessibility, external development, and optically clear embryo are just a few of the features that set the zebrafish apart as a particularly well-suited model for studying vascular development. However, there is little precedent for its use as a tool for the experimental study of therapeutic angiogenesis. Here, we review the use of the zebrafish for studying vascular development and patterning, and discuss how the zebrafish might be used more directly as a model for developing and testing effective therapeutic angiogenesis approaches. PMID- 14534145 TI - Myoblast-mediated gene transfer for therapeutic angiogenesis and arteriogenesis. AB - Therapeutic angiogenesis aims at generating new blood vessels by delivering growth factors such as VEGF and FGF. Clinical trials are underway in patients with peripheral vascular and coronary heart disease. However, increasing evidence indicates that the new vasculature needs to be stabilized to avoid deleterious effects such as edema and hemangioma formation. Moreover, a major challenge is to induce new vessels that persist following cessation of the angiogenic stimulus. Mature vessels may be generated by modulating timing and dosage of growth factor expression, or by combination of 'growth' factors with 'maturation' factors like PDGF-BB, angiopoietin-1 or TGF-beta. Myoblast-mediated gene transfer has unique characteristics that make it a useful tool for studying promising novel approaches to therapeutic angiogenesis. It affords robust and long-lasting expression, and can be considered as a relatively rapid form of 'adult transgenesis' in muscle. The combined insertion of different gene constructs into single myoblasts and their progeny allows the simultaneous expression of different 'growth' and 'maturation' factors within the same cell in vivo. The additional insertion of a reporter gene makes it possible to analyze the phenotype of the vessels surrounding the transgenic muscle fibers into which the myoblasts have fused. The effects of timing and duration of gene expression can be studied by using tetracycline-inducible constructs, and dosage effects by selecting subpopulations consistently expressing distinct levels of growth factors. Finally, the autologous cell-based approach using transduced myoblasts could be an alternative gene delivery system for therapeutic angiogenesis in patients, avoiding the toxicities seen with some viral vectors. PMID- 14534146 TI - Designer blood vessels and therapeutic revascularization. AB - Inadequate vascular perfusion leads to fatal heart attacks, chronic ulcers, and other serious clinical conditions. The body's capacity to restore vascular perfusion through angiogenesis and arteriogenesis is often impaired by pre existing disease, and availability of native replacements for nonfunctional arteries is limited in many patients. Thus, recreating blood vessels of various calibres through novel engineering technologies has emerged as a radical option among therapeutic strategies for revascularization. Ranging from artificial, recycled or reassembled natural conduits to sophisticated microdevices, we refer to these as 'designer blood vessels'. Our common efforts to continuously improve vascular replacement design have provided many clues about our own blood vessels, but nature's ability to create nonthrombogenic, immunocompatible, strong, yet biologically responsive blood vessels remains unparalleled. Just as art reproductions never equal the original masterpiece, designer blood vessels may never attain nature's perfection. Nevertheless, they will provide a valuable option as long as they come close enough and are available to many. PMID- 14534147 TI - Testing clinical therapeutic angiogenesis using basic fibroblast growth factor (FGF-2). AB - Therapeutic angiogenesis represents an attempt to relieve inadequate blood flow by the directed growth and proliferation of blood vessels. Neovascularization is a complex process involving multiple growth factors, receptors, extracellular matrix glycoproteins, intracellular and extracellular signaling pathways, and local and bone-marrow-derived constituent cells, all responding to a symphonic arrangement of temporal and spatial cues. In cardiovascular disease, patients with refractory angina and lower extremity intermittent claudication seem most amenable to early tests of therapeutic angiogenesis. Monotherapy with the recombinant protein basic fibroblast growth factor (FGF-2) has been tested in six human trials. These have shown provisional safety, and two have provided 'proof of concept' for the strategy of therapeutic angiogenesis. One large randomized phase II trial failed to show significant efficacy in coronary artery disease. Another showed significant efficacy in peripheral artery disease, although the magnitude of benefit was disappointing at the dose tested. This overview details the suitable clinical trial design and further steps toward the clinical development of FGF-2. PMID- 14534148 TI - Regulation of leptin distribution between plasma and cerebrospinal fluid by cholecystokinin receptors. AB - Cholecystokinin (CCK) is a postprandial hormone that elicits a satiating effect and regulates feeding behaviour. CCK has been shown to enhance the effect of leptin in several experimental paradigms. The goal of this work was to characterize the effect of endogenous CCK on plasma leptin content by using CCK receptor antagonists. Therefore, we administered SR-27897, a selective CCK1 receptor antagonist, and L-365260, a selective CCK2 receptor antagonist, to fed and food-deprived rats and determined plasma leptin concentration by enzyme immunoassay. Plasma insulin and glucose concentration as well as food intake were also determined. Under our conditions, SR-27897 increased plasma concentration of leptin both in fed and food-deprived rats. It also increased food intake as well as plasma concentration of insulin in fed animals. L-365260 increased plasma leptin concentration only in fed rats. In animals receiving exogenous leptin, CCK 8 increased the ratio between the concentration of leptin in cerebrospinal fluid and plasma. These results show that CCK receptor antagonists increases plasma concentration of leptin and suggest that endogenous CCK may facilitate the uptake of plasma leptin to the cerebrospinal fluid. PMID- 14534149 TI - Inhibition of cardiac sodium currents by toluene exposure. AB - Toluene is an industrial solvent widely used as a drug of abuse, which can produce sudden sniffing death due to cardiac arrhythmias. In this paper, we tested the hypothesis that toluene inhibits cardiac sodium channels in Xenopus laevis oocytes transfected with Nav1.5 cDNA and in isolated rat ventricular myocytes. In oocytes, toluene inhibited sodium currents (INa+) in a concentration dependent manner, with an IC50 of 274 microm (confidence limits: 141-407 microm). The inhibition was complete, voltage-independent, and slowly reversible. Toluene had no effect on: (i). the shape of the I-V curves; (ii). the reversal potential of Na+; and (iii). the steady-state inactivation. The slow recovery time constant from inactivation of INa+ decreased with toluene exposure, while the fast recovery time constant remained unchanged. Block of INa+ by toluene was use- and frequency-dependent. In rat cardiac myocytes, 300 microm toluene inhibited the sodium current (INa+) by 62%; this inhibition was voltage independent. These results suggest that toluene binds to cardiac Na+ channels in the open state and unbinds either when channels move between inactivated states or from an inactivated to a closed state. The use- and frequency-dependent block of INa+ by toluene might be responsible, at least in part, for its arrhythmogenic effect. PMID- 14534150 TI - Ginsenoside Rg3 inhibits phenylephrine-induced vascular contraction through induction of nitric oxide synthase. AB - Ginsenoside Rg3 (Rg3) isolated from Panax ginseng relaxes vessels and exerts a cytoprotective effect. In view of the fact that nitric oxide (NO) is involved in vascular hyporeactivity and immunostimulation, the effects of total ginsenosides (GS) and Rg3 on the vascular responses and the expression of inducible nitric oxide synthase (iNOS) were investigated. Vasocontraction of endothelium-denuded aortic ring was induced by phenylephrine with or without GS or Rg3. The expression of iNOS was assessed by Western blot and RT-PCR analyses. NF-kappaB activation was monitored by gel shift, immunoblot and immunocytochemical analyses. Incubation of the endothelium-denuded aortic ring with GS or Rg3 inhibited phenylephrine-induced vasocontraction, which was abrogated by NOS inhibition. GS or Rg3 increased NO production in aortic rings, but Rb1, Rc, Re and Rg1 had no effect. Aortic rings obtained from rats treated with GS or Rg3 responded to phenylnephrine to a lesser extent, while producing NO to a larger extent, than those from control animals. GS or Rg3 induced iNOS in vascular smooth muscle. Rg3 induced iNOS with increase in NO production in Raw264.7 cells. Rg3 increased NF-kappaB DNA binding, whose band was supershifted with anti-p65 and anti-p50 antibodies, and elicited p65 nuclear translocation, which was accompanied by phosphorylation and degradation of I-kappaBalpha. PKC regulated iNOS induction by Rg3. In conclusion, Rg3 relaxes vessels as a consequence of NO production, to which iNOS induction contributes, and iNOS induction by Rg3 accompanied NF-kappaB activation, which involves phosphorylation and degradation of I-kappaBalpha and nuclear translocation of p65. PMID- 14534151 TI - Mechanisms involved in the early increase of serotonin contraction evoked by endotoxin in rat middle cerebral arteries. AB - The present study investigated the mechanisms involved in the increased 5 hydroxytryptamine (5-HT) vasoconstriction observed in rat middle cerebral arteries exposed in vitro to lipopolysaccharide (LPS, 10 microg x ml-1) for 1-5 h. Functional, immunohistochemical and Western blot analysis and superoxide anion measurements by ethidium fluorescence were performed. LPS exposure increased 5-HT (10 microm) vasoconstriction only during the first 4 h. In contrast to control tissue, indomethacin (10 microm), the COX-2 inhibitor NS 398 (10 microm), the TXA2/PGH2 receptor antagonist SQ 29548 (1 microm) and the TXA2 synthase inhibitor furegrelate (1 microm) reduced 5-HT contraction of LPS-treated arteries from hour one. The iNOS inhibitor aminoguanidine (0.1 mm) increased 5-HT contraction from hour three of LPS incubation. The superoxide anion scavenger superoxide dismutase (SOD, 100 U ml-1) and the H2O2 scavenger catalase (1000 U ml-1), as well as the respective inhibitors of NAD(P)H oxidase and xanthine oxidase, apocynin (0.3 mm) and allopurinol (0.3 mm), reduced 5-HT contraction after LPS incubation. LPS induced an increase in superoxide anion levels that was abolished by PEG-SOD. Subthreshold concentrations of the TXA2 analogue U 46619, xanthine/xanthine oxidase and H2O2 potentiated, whereas those of sodium nitroprusside inhibited, the 5-HT contraction. COX-2 expression was increased at 1 and 5 h of LPS incubation, while that of iNOS, Cu/Zn-SOD and Mn-SOD was only increased after 5 h. All the three vascular layers expressed COX-2 and Cu/Zn-SOD. iNOS expression was detected in the endothelium and adventitia after LPS. In conclusion, increased production of TXA2 from COX-2, superoxide anion and H2O2 enhanced vasoconstriction to 5-HT during the first few hours of LPS exposure; iNOS and SOD expression counteracted that increase at 5 h. These changes can contribute to the disturbance of cerebral blood flow in endotoxic shock. PMID- 14534152 TI - Pharmacological characterization of a 7-benzylidenenaltrexone-preferring opioid receptor in porcine ileal submucosa. AB - In the intestine, opioids produce antidiarrhoeal and constipating actions that are mediated by enteric neurones. Through interactions with opioid receptors (ORs) on submucosal neurones, opioids suppress active ion transport evoked by transmural electrical stimulation (TES) in mucosa-submucosa sheets from the porcine ileum. In this study, we examined the pharmacological characteristics of the previously described OR, which is sensitive to the delta1-OR antagonist 7 benzylidenenaltrexone and modulates neurogenic transepithelial ion transport in this tissue preparation. Increases in short-circuit current (Isc, a measure of active anion transport) evoked by TES in ileal mucosa-submucosa sheets were inhibited by opioid agonists possessing high selectivity for either delta- or micro-ORs including [d-Pen2,5]enkephalin (DPDPE), [d-Ala2, Glu4]deltorphin II, and [d-Ala2, N-Me-Phe4, Gly5-ol]enkephalin (DAMGO). As determined by the Schild analysis, the actions of these agonists were competitively inhibited by 7 benzylidenenaltrexone. The nonequilibrium micro-OR antagonist beta-funaltrexamine inhibited the actions of DAMGO only at a high concentration (1 microm) but did not alter DPDPE or deltorphin II action. At concentrations up to 10 microm, the nonequilibrium delta-OR antagonist naltrindole 5'-isothiocyanate did not alter the actions of delta- or micro-OR agonists. Radioligand binding analyses of neuronal homogenates from the ileal submucosa revealed that the nonselective OR ligand [3H]diprenorphine bound to two populations of specific binding sites. One of these sites possessed binding characteristics similar to the delta-OR. In summary, neurogenic ion transport in the porcine intestine is modulated by an OR which shares pharmacological characteristics of both micro- and delta-ORs and may represent a novel receptor entity. PMID- 14534154 TI - Vanilloid type 1 receptors (VR1) on trigeminal sensory nerve fibres play a minor role in neurogenic dural vasodilatation, and are involved in capsaicin-induced dural dilation. AB - Capsaicin, the active substance in chilli peppers, activates the vanilloid type 1 receptor (VR1) rather than the vanilloid-like receptor (VRL1) in the trigeminal ganglion and nucleus of small and medium C- and Adelta-fibres. Capsaicin induces calcitonin gene-related peptide (CGRP) release when VR1 receptors are activated, and this can be reversed by both the VR1 receptor antagonist capsazepine and the CGRP blocker alphaCGRP8-37 in vitro. In this study we used intravital microscopy to look at the possible role of the VR1 receptor in the trigeminovascular system in producing dilation of dural blood vessels. Capsazepine (3 mg kg-1) was given to study the effect of the VR1 receptor in dural vessel dilation produced by either electrical stimulation, CGRP (1 microg x kg-1) or capsaicin (7 microg x kg 1) bolus injection. We also looked at the effect of the CGRP blocker alphaCGRP8 37 (300 microg x kg-1) on capsaicin-induced dilation so that we could see if the results found in vitro could also be found in vivo. Electrical stimulation of the dura mater produced a robust vasodilator response between 130 and 137% of baseline diameter that was no different across four repeat stimuli (F3,18=0.6, P=0.61). CGRP similarly produced a dilatation of 99-111% that was no different across four baseline infusions (F3,15=2.4, P=0.113). Capsaicin also produced a consistent dilation of between 112 and 120% of baseline across three injections (F2,10=0.6, P=0.567). Capsazepine did not inhibit the dilation brought about by either electrical stimulation or CGRP injection, while it was able to inhibit the dilation brought about by capsaicin (t5=3.4, P<0.05). AlphaCGRP8-37 also inhibited the capsaicin-induced dilation (t5=7.4, P<0.05) probably inhibiting the action of released CGRP at the CGRP receptor. The study demonstrates that capsaicin can repeatedly induce dural vessel dilation in vivo, presumably through inducing CGRP release from trigeminal sensory nerve fibres, while C-fibres may have been desensitised. The data imply that the VR1 receptor plays only a minor role in trigeminovascular-induced dural vessel dilation. PMID- 14534153 TI - Chronic oral supplementation with sepiapterin prevents endothelial dysfunction and oxidative stress in small mesenteric arteries from diabetic (db/db) mice. AB - We previously reported that acute incubation with tetrahydrobiopterin (BH4) or sepiapterin, a cofactor for endothelial nitric oxide synthase and a stable precursor of BH4, respectively, enhanced the acetylcholine (Ach)-induced relaxation of isolated small mesenteric arteries (SMA) from diabetic (db/db) mice. In this study, we investigated the effect of chronic oral supplementation of sepiapterin (10 mg x kg-1 x day-1) to db/db mice on endothelium function, biopterin levels and lipid peroxidation in SMA. Oral dietary supplementation with sepiapterin had no effect on glucose, triglyceride, cholesterol levels and body weight. SMA from db/db mice showed enhanced vascular reactivity to phenylephrine, which was corrected with sepiapterin supplementation. Furthermore, Ach, but not sodium nitroprusside-induced relaxation, was improved with sepiapterin supplementation in db/db mice. BH4 levels and guanosine triphosphate cyclohydrolase I activity in SMA were similar in db/+ and db/db mice. Sepiapterin treatment had no effects on BH4 or guanosine triphosphate cyclohydrolase I activity. However, the level of dihydrobiopterin+biopterin was higher in SMA from db/db mice, which was corrected following sepiapterin treatment. Thiobarbituric acid reactive substance, malondialdehyde, a marker of lipid peroxidation, was higher in SMA from db/db mice, and was normalized by sepiapterin treatment. These results indicate that sepiapterin improves endothelial dysfunction in SMA from db/db mice by reducing oxidative stress. Furthermore, these results suggest that decreased biosynthesis of BH4 may not be the basis for endothelial dysfunction in SMA from db/db mice. PMID- 14534155 TI - In vitro inhibition of human neutrophil histotoxicity by ambroxol: evidence for a multistep mechanism. AB - Neutrophils are major culprits for the protease/antiprotease imbalance during various lung diseases, that is, chronic obstructive pulmonary disease, cystic fibrosis, idiopathic pulmonary fibrosis and adult respiratory distress syndrome. Thus, these cells are presently considered an ideal target for the pharmacologic control of tissue injury during these diseases. This study was planned in order to investigate if ambroxol and its precursor bromhexine are actually capable of preventing alpha-1-antitrypsin (A1AT) inactivation by stimulated neutrophils and possibly to look into the mechanisms underlying this event. Ambroxol inhibited the production of superoxide anion by activated neutrophils, whereas bromhexine had no inhibitory effect. Ambroxol decreased the production of hypochlorous acid (HOCl) from activated neutrophils with high efficiency, whereas bromhexine had a modest activity. Ambroxol and bromhexine were capable of limiting the chlorination of monochlorodimedon by HOCl, displaying the capacity of directly scavenging the oxidant. Ambroxol decreased the release of elastase and myeloperoxidase from activated neutrophils, whereas bromhexine was ineffective. Ambroxol prevented the A1AT inactivation by neutrophils, whereas bromhexine was completely ineffective. Among drugs currently available for in vivo use in humans, ambroxol is unique by virtue of its ability to prevent neutrophil mediated A1AT inactivation via inhibition of HOCl production as well as HOCl scavenging. Also taking into account its capacity for curbing elastase release, the drug displays the potential to lessen the burden of oxidants/proteases and to increase the antiprotease shield at the site of inflammation. Thus, ambroxol appears to be a good candidate for raising attempts to develop new therapeutic histoprotective approaches to inflammatory bronchopulmonary diseases. PMID- 14534156 TI - Roles of KATP channels in delayed cardioprotection and intracellular Ca(2+) in the rat heart as revealed by kappa-opioid receptor stimulation with U50488H. AB - The effect of preconditioning with U50488 H (UP), a selective kappa-opioid receptor (kappa-OR) agonist, on infarct size and intracellular Ca2+ ([Ca2+]i) in the heart subjected to ischaemic insults were studied and evaluated. U50488 H administered intravenously reduced the infarct size 18-48 h after administration in isolated hearts subjected to regional ischaemia/reperfusion (I/R). The effect was dose dependent. A peak effect was reached at 10 mg x kg-1 U50488 H and at 24 h after administration. The effect of 10 mg x kg-1 U50488 H at 24 h after administration was abolished by nor-binaltorphimine (nor-BNI), a selective kappa OR antagonist, indicating the effect was kappa-OR mediated. The infarct reducing effect of U50488 H was attenuated when a selective blocker of mitochondrial (5 hydroxydecanoic acid, 5-HD) or sarcolemmal (HRM-1098) ATP-sensitive potassium channel (KATP) was coadministered with U50488 H 24 h before ischaemia or when 5 HD was administered just before ischaemia. U50488 H also attenuated the elevation in [Ca2+]i and reduction in electrically induced [Ca2+]i transient in cardiomyocytes subjected to ischaemic insults. The effects were reversed by blockade of KATP channel, which abolished the protective effect of preconditioning with U50488 H. The results indicated that mitochondrial KATP channel serves as both a trigger and a mediator, while sarcolemmal KATP channel as a trigger only, of delayed cardioprotection of kappa-OR stimulation. The effects of these channels may result from prevention/attenuation of [Ca2+]i overload induced by ischaemic insults. PMID- 14534157 TI - KCNQ2 and KCNQ3 potassium channel genes in benign familial neonatal convulsions: expansion of the functional and mutation spectrum. AB - Benign familial neonatal convulsions (BFNC) is a rare autosomal dominant generalized epilepsy of the newborn infant. Seizures occur repeatedly in the first days of life and remit by approximately 4 months of age. Previously our laboratory cloned two novel potassium channel genes, KCNQ2 and KCNQ3, and showed that they are mutated in patients with BFNC. In this report, we characterize the breakpoints of a previously reported interstitial deletion in the KCNQ2 gene and show that only KCNQ2 is deleted. We identify 11 novel mutations in KCNQ2 and one novel mutation in the KCNQ3 potassium channel genes. In one family, the phenotype extends beyond neonatal seizures and includes rolandic seizures, and a subset of families has onset of seizures in infancy. In the Xenopus oocyte expression system, we characterize five KCNQ2 and one KCNQ3 disease-causing mutations. These mutations cause a variable loss of function, and selective effects on the biophysical properties of KCNQ2/KCNQ3 heteromultimeric channels. We report here the first dominant negative mutation in KCNQ2 that has a phenotype of neonatal seizures without permanent clinical CNS impairment. PMID- 14534158 TI - Gradual loss of myelin and formation of an astrocytic scar during Wallerian degeneration in the human spinal cord. AB - Axons undergo Wallerian degeneration distal to a point of injury. Experimental investigations have documented many of the cellular and molecular events that underlie this behaviour. Since relatively little is known about such events in human CNS pathologies and current experimental intervention strategies indicate the possibility of significant axon regeneration along the original degenerated fibre tract, we performed an immunohistochemical investigation of the dynamics of Wallerian degeneration in post mortem spinal cords of patients who died 2 days to 30 years after either cerebral infarction or traumatic spinal cord injury. Neurofilament (NF) staining demonstrated a spatio-temporal pattern of axonal loss within degenerating descending nerve fibre tracts that could be detected close to the lesion as early as 12 days after injury and progressed to an almost complete loss of NF immunoreactivity at survival times of 1 year and longer. Immunohistochemistry for glial fibrillary acidic protein revealed a late astrocytic reaction starting at 4 months after injury in the degenerating tracts, leading to the long-term deposition of a dense astrocytic scar. These events were accompanied by the gradual reduction of myelin basic protein in affected nerve fibre tracts, leading to almost complete loss by 3 years after injury. Since the extracellular matrix molecule chondroitin sulphate proteoglycan (CSPG) is known to be strongly inhibitory for axonal regeneration and to be a major component of gliotic scar tissues, we investigated the possible deposition of CSPG within the degenerating nerve fibre tracts. Apart from a local up-regulation close to the lesion site, our results show no enhanced CSPG expression within degenerated tracts at any survival time. This suggests that despite the apparent lack of CSPG in Wallerian degeneration, the slow reduction of CNS myelin and the long-term deposition of a dense astrocytic scar may present an environment that is non supportive for axon regrowth. PMID- 14534159 TI - Histopathology and reorganization of chandelier cells in the human epileptic sclerotic hippocampus. AB - Impairment of GABA-mediated inhibition is one of the main hypotheses invoked to explain seizure activity, both in experimental models and in human epilepsy. We have studied the distribution and the neurochemical characteristics of certain GABAergic circuits in the normal and epileptic human sclerotic hippocampal formation. We have focused our attention mainly on chandelier cells because, together with basket cells, they are considered to have powerful effects on spike generation. Chandelier cells represent a unique type of interneuron whose axon terminals (Ch-terminals) form synapses with the axon initial segments of cortical pyramidal cells and granular cells of the dentate gyrus. Different neurochemical subpopulations of chandelier cells have been identified by immunocytochemistry, mainly in the neocortex. Markers for Ch-terminals include the GABA transporter 1 (GAT-1), the polysialylated form of the cell-surface glycoprotein neural cell adhesion molecule (PSA-NCAM) and the calcium-binding proteins parvalbumin (PV) and calbindin D-28k (CB). In the normal hippocampal formation, GAT-1- and PV immunoreactive (-ir) Ch-terminals were identified in the granular and polymorphic layers of the dentate gyrus, in the strata pyramidale and oriens of the CA fields, and in the pyramidal layer of the subicular complex. In addition, and in contrast to the hippocampus and dentate gyrus, subsets of Ch-terminals in the upper pyramidal layer of the normal subiculum express CB and PSA-NCAM. The sclerotic hippocampus of epileptic patients presented an impressive morphological and neurochemical reorganization of Ch-terminals and basket formations. This was apparent in the dentate gyrus and hippocampal formation, but not in the subiculum, which appeared to remain unaltered. Principally, numerous and more complex PV- and CB-ir Ch-terminals, as well as dense PV-ir basket formations, appeared in some hippocampal segments, whereas in other regions there was a lack of labelled elements. These changes varied considerably not only between different patients, but also within different hippocampal fields in a given patient. In general, the changes were not correlated with the clinical characteristics or degree of histopathological alterations observed in the patients, such as granular cell dispersion, neuron loss and proliferation of mossy fibres. However, some surviving neurons in the regions adjacent to the areas of neuron loss were consistently innervated by dense basket formations and complex Ch-terminals. These results indicate that, in the human epileptic hippocampus, GABAergic circuits are more highly modified than previously thought. When considered along with other extrahippocampal alterations, we suggest that these changes are important in the pathophysiology of temporal lobe epilepsy associated with hippocampal sclerosis. PMID- 14534160 TI - Minute quantities of misfolded mutant superoxide dismutase-1 cause amyotrophic lateral sclerosis. AB - Mutant forms of superoxide dismutase-1 (SOD1) cause amyotrophic lateral sclerosis (ALS) by an unknown noxious mechanism. Using an antibody against a novel epitope in the G127insTGGG mutation, mutant SOD1 was studied for the first time in spinal cord and brain of an ALS patient. The level was below 0.5% of the SOD1 level in controls. In corresponding transgenic mice the content of mutant SOD1 was also low, although it was enriched in spinal cord and brain compared with other tissues. In the mice the misfolded mutant SOD1 aggregated rapidly and 20% occurred in steady state as detergent-soluble protoaggregates. The misfolded SOD1 and the protoaggregates form, from birth until death, a potentially noxious burden that may induce the motor neuron injury. Detergent-resistant aggregates, as well as inclusions of mutant SOD1 in motor neurons and astrocytes, accumulated in spinal cord ventral horns of the patient and mice with terminal disease. The inclusions and aggregates may serve as terminal markers of long-term assault by misfolded SOD1 and protoaggregates. PMID- 14534164 TI - Computational detection of cis -regulatory modules. AB - MOTIVATION: The transcriptional regulation of a metazoan gene depends on the cooperative action of multiple transcription factors that bind to cis-regulatory modules (CRMs) located in the neighborhood of the gene. By integrating multiple signals, CRMs confer an organism specific spatial and temporal rate of transcription. RESULTS: Based on the hypothesis that genes that are needed in exactly the same conditions might share similar regulatory switches, we have developed a novel methodology to find CRMs in a set of coexpressed or coregulated genes. The ModuleSearcher algorithm finds for a given gene set the best scoring combination of transcription factor binding sites within a sequence window using an A(*)procedure for tree searching. To keep the level of noise low, we use DNA sequences that are most likely to contain functional cis-regulatory information, namely conserved regions between human and mouse orthologous genes. The ModuleScanner performs genomic searches with a predicted CRM or with a user defined CRM known from the literature to find possible target genes. The validity of a set of putative targets is checked using Gene Ontology annotations. We demonstrate the use and effectiveness of the ModuleSearcher and ModuleScanner algorithms and test their specificity and sensitivity on semi-artificial data. Next, we search for a module in a cluster of gene expression profiles of human cell cycle genes. AVAILABILITY: The ModuleSearcher is available as a web service within the TOUCAN workbench for regulatory sequence analysis, which can be downloaded from http://www.esat.kuleuven.ac.be/~dna/BioI. PMID- 14534161 TI - Metabolic changes and electro-clinical patterns in mesio-temporal lobe epilepsy: a correlative study. AB - Interictal hypometabolism is commonly found in mesio-temporal lobe epilepsy (MTLE), but its pathophysiology remains incompletely understood. We hypothesized that metabolic changes reflect the preferential networks involved by ictal discharges. We analysed the topography of interictal hypometabolism according to electro-clinical patterns in 50 patients with unilateral hippocampal sclerosis (HS) and consistent features of MTLE. Based on electro-clinical correlations, we identified four groups: (i) mesial group (13 cases) characterized by mesial seizure onset without evidence of early spread beyond the temporal lobe; (ii) anterior mesio-lateral group (AML; 18 cases) with early anterior spread involving the anterior lateral temporal cortex and insulo-fronto-opercular areas; (iii) widespread mesio-lateral group (WML; 15 cases) with wide spread (involving both anterior and posterior lateral temporal and perisylvian areas); and (iv) bitemporal (BT) group (four cases) with early contralateral temporal spread. Results of [18F]fluorodeoxyglucose-PET imaging in each group were compared with those of 10 control subjects using statistical parametric mapping software (SPM99). MRI data and surgical outcome in each group were compared with metabolic findings. Hypometabolism was limited to hippocampal gyrus, temporal pole and insula in the mesial group. Gradual involvement of lateral temporal cortex, insula and perisylvian areas was observed in the AML and WML groups. The BT group differed from the others with mild bitemporal involvement, bilateral insular hypometabolism and longer epilepsy duration. MRI structural abnormalities outside of the mesial formations were detected in 65% of the cases. Neither the severity of HS nor temporal atrophy appeared related to the topography of hypometabolism. However, temporal hypometabolism was more extended when temporo-polar signal changes were detected. Among operated patients (n = 43), seizure-free outcome was obtained in 82%. Surgical outcome appeared more favourable in the mesial group. However, the difference between the four groups was not significant. Our results suggest that hypometabolism in MTLE may be related to ictal discharge generation and spread pathways, even if structural changes and epilepsy duration may also play a role. PMID- 14534166 TI - Searching for statistically significant regulatory modules. AB - MOTIVATION: The regulatory machinery controlling gene expression is complex, frequently requiring multiple, simultaneous DNA-protein interactions. The rate at which a gene is transcribed may depend upon the presence or absence of a collection of transcription factors bound to the DNA near the gene. Locating transcription factor binding sites in genomic DNA is difficult because the individual sites are small and tend to occur frequently by chance. True binding sites may be identified by their tendency to occur in clusters, sometimes known as regulatory modules. RESULTS: We describe an algorithm for detecting occurrences of regulatory modules in genomic DNA. The algorithm, called mcast, takes as input a DNA database and a collection of binding site motifs that are known to operate in concert. mcast uses a motif-based hidden Markov model with several novel features. The model incorporates motif-specific p-values, thereby allowing scores from motifs of different widths and specificities to be compared directly. The p-value scoring also allows mcast to only accept motif occurrences with significance below a user-specified threshold, while still assigning better scores to motif occurrences with lower p-values. mcast can search long DNA sequences, modeling length distributions between motifs within a regulatory module, but ignoring length distributions between modules. The algorithm produces a list of predicted regulatory modules, ranked by E-value. We validate the algorithm using simulated data as well as real data sets from fruitfly and human. AVAILABILITY: http://meme.sdsc.edu/MCAST/paper PMID- 14534167 TI - Ab initio reconstruction of metabolic pathways. AB - We propose a new formulation for the problem of ab initio metabolic pathway reconstruction. Given a set of biochemical reactions together with their substrates and products, we consider the reactions as transfers of atoms between the chemical compounds and we look for successions of reactions transferring a maximal (or preset) number of atoms between a given source and sink compound. We state this problem as the one of finding a composition of partial injections that maximizes the image size. First, we study the theoretical complexity of this problem, state some related problems and then give a practical algorithm to solve them. Finally, we present two applications of this approach to the reconstruction of the tryptophan biosynthesis pathway and to the glycolysis. PMID- 14534169 TI - HMM sampling and applications to gene finding and alternative splicing. AB - The standard method of applying hidden Markov models to biological problems is to find a Viterbi (maximal weight) path through the HMM graph. The Viterbi algorithm reduces the problem of finding the most likely hidden state sequence that explains given observations, to a dynamic programming problem for corresponding directed acyclic graphs. For example, in the gene finding application, the HMM is used to find the most likely underlying gene structure given a DNA sequence. In this note we discuss the applications of sampling methods for HMMs. The standard sampling algorithm for HMMs is a variant of the common forward-backward and backtrack algorithms, and has already been applied in the context of Gibbs sampling methods. Nevetheless, the practice of sampling state paths from HMMs does not seem to have been widely adopted, and important applications have been overlooked. We show how sampling can be used for finding alternative splicings for genes, including alternative splicings that are conserved between genes from related organisms. We also show how sampling from the posterior distribution is a natural way to compute probabilities for predicted exons and gene structures being correct under the assumed model. Finally, we describe a new memory efficient sampling algorithm for certain classes of HMMs which provides a practical sampling alternative to the Hirschberg algorithm for optimal alignment. The ideas presented have applications not only to gene finding and HMMs but more generally to stochastic context free grammars and RNA structure prediction. PMID- 14534170 TI - Predicting gene function in Saccharomyces cerevisiae. AB - MOTIVATION: S.cerevisiae is one of the most important model organisms, and has has been the focus of over a century of study. In spite of these efforts, 40% of its open reading frames (ORFs) remain classified as having unknown function (MIPS: Munich Information Center for Protein Sequences). We wished to make predictions for the function of these ORFs using data mining, as we have previously successfully done for the genomes of M.tuberculosis and E.coli. Applying this approach to the larger and eukaryotic S.cerevisiae genome involves modifying the machine learning and data mining algorithms, as this is a larger organism with more data available, and a more challenging functional classification. RESULTS: Novel extensions to the machine learning and data mining algorithms have been devised in order to deal with the challenges. Accurate rules have been learned and predictions have been made for many of the ORFs whose function is currently unknown. The rules are informative, agree with known biology and allow for scientific discovery. AVAILABILITY: All predictions are freely available from http://www.genepredictions.org, all datasets used in this study are freely available from http://www.aber.ac.uk/compsci/Research/bio/dss/yeastdataand software for relational data mining is available from http://www.aber.ac.uk/compsci/Research/bio/dss/polyfarm. PMID- 14534171 TI - Exploring potential target genes of signaling pathways by predicting conserved transcription factor binding sites. AB - Many cellular signaling pathways induce gene expression by activating specific transcription factor complexes. Conventional approaches to the prediction of transcription factor binding sites lead to a notoriously high number of false discoveries. To alleviate this problem, we consider only binding sites that are conserved in man-mouse genomic sequence comparisons. We employ two alternative methods for predicting binding sites: exact matches to validated binding site sequences and weight matrix scans. We then ask the question whether there is a characteristic association between a transcription factor or set thereof to a particular group of genes. Our approach is tested on genes, which are induced in dendritic cells in response to the cells' exposure to LPS. We chose this example because the underlying signaling pathways are well understood. We demonstrate the benefit of conserved predicted binding sites in interpreting the LPS experiment. Additionally, we find that both methods for the prediction of conserved binding sites complement one another. Finally, our results suggest a distinct role for SRF in the context of LPS-induced gene expression. PMID- 14534173 TI - Inferring strengths of protein-protein interactions from experimental data using linear programming. AB - MOTIVATION: Several computational methods have been proposed for inference of protein-protein interactions. Most of the existing methods assume that protein protein interaction data are given as binary data (i.e. whether or not each protein pair interacts). However, multiple biological experiments are performed for the same protein pairs and thus the ratio (strength) of the number of observed interactions to the number of experiments is available for each protein pair. RESULTS: We propose a new method for inference of protein-protein interactions from such experimental data. This method tries to minimize the errors between the ratios of observed interactions and the predicted probabilities in training data, where this problem is formalized as a linear program based on a probabilistic model. We compared the proposed method with the association method, the EM method and the SVM-based method using real interaction data. It is shown that a variant of the method is comparable to existing methods for binary data. It is also shown that the method outperforms existing methods for numerical data. AVAILABILITY: Programs transforming input data into LP format files are available upon request. PMID- 14534174 TI - Upper bounds on maximum likelihood for phylogenetic trees. AB - We introduce a mechanism for analytically deriving upper bounds on the maximum likelihood for genetic sequence data on sets of phylogenies. A simple 'partition' bound is introduced for general models. Tighter bounds are developed for the simplest model of evolution, the two state symmetric model of nucleotide substitution under the molecular clock. This follows earlier theoretical work which has been restricted to this model by analytic complexity. A weakness of current numerical computation is that reported 'maximum likelihood' results cannot be guaranteed, both for a specified tree (because of the possibility of multiple maxima) or over the full tree space (as the computation is intractable for large sets of trees). The bounds we develop here can be used to conclusively eliminate large proportions of tree space in the search for the maximum likelihood tree. This is vital in the development of a branch and bound search strategy for identifying the maximum likelihood tree. We report the results from a simulation study of approximately 10(6) data sets generated on clock-like trees of five leaves. In each trial a likelihood value of one specific instance of a parameterised tree is compared to the bound determined for each of the 105 possible rooted binary trees. The proportion of trees that are eliminated from the search for the maximum likelihood tree ranged from 92% to almost 98%, indicating a computational speed-up factor of between 12 and 44. PMID- 14534175 TI - A rapid method for detection of putative RNAi target genes in genomic data. AB - RNAi, inhibition of gene expression by double stranded RNA molecules, has rapidly become a powerful laboratory technique to study gene function. The effectiveness of the procedure raised the question of whether this laboratory technique may actually mimic a natural cellular control mechanism that works on similar principles. Indeed recent evidence is accumulating to suggest that RNAi is a natural control mechanism that might even serve as a primitive immune response against RNA viruses and retroposons. Three different interference scenarios seem to be utilized by various RNAi mechanisms. One of the mechanisms involves degradation of mRNA molecules. Here we suggest a method to systematically scan entire genomes simultaneously for RNAi elements and the presence of cellular genes that are degraded by these RNAi elements via exact short base-pair matching. The method is based on scanning the genomes using a suffix tree data structure that was specifically modified to identify sets of combinations of repeated and inverted repeated sequences of 20 bp or more. Initial scan suggest that a large number, about 7% of C.elegans and 3% of C.briggsae genes, have the potential to be subject to natural RNAi control. Two methods are proposed to further analyze these genes to select the cases that are more likely to be actual cases of RNAi control. One method involves looking for ESTs that can provide direct evidence that RNAi control element are indeed expressed. The other method looks for synteny between C.elegans and C.briggsae assuming that genes that might be under RNAi control in both organisms are more likely to be biological significant. Taken together, supportive evidence was found for about 70 genes to be under RNAi control. Among these genes are: transposase, hormone receptors, homeobox proteins, defensin, actins, and several types of collagens. While our method is not capable of detecting all cases of natural RNAi control, it points to a large number of potential cases that can be further verified by experimental work. PMID- 14534176 TI - A novel approach to fold recognition using sequence-derived properties from sets of structurally similar local fragments of proteins. AB - Comparative modeling methods can consistently produce reliable structural models for protein sequences with more than 25% sequence identity to proteins with known structure. However, there is a good chance that also sequences with lower sequence identity have their structural components represented in structural databases. To this end, we present a novel fragment-based method using sets of structurally similar local fragments of proteins. The approach differs from other fragment-based methods that use only single backbone fragments. Instead, we use a library of groups containing sets of sequence fragments with geometrically similar local structures and extract sequence related properties to assign these specific geometrical conformations to target sequences. We test the ability of the approach to recognize correct SCOP folds for 273 sequences from the 49 most popular folds. 49% of these sequences have the correct fold as their top prediction, while 82% have the correct fold in one of the top five predictions. Moreover, the approach shows no performance reduction on a subset of sequence targets with less than 10% sequence identity to any protein used to build the library. PMID- 14534178 TI - Discovery of significant rules for classifying cancer diagnosis data. AB - METHODS AND RESULTS: We introduce a new method to discover many diversified and significant rules from high dimensional profiling data. We also propose to aggregate the discriminating power of these rules for reliable predictions. The discovered rules are found to contain low-ranked features; these features are found to be sometimes necessary for classifiers to achieve perfect accuracy. The use of low-ranked but essential features in our method is in contrast to the prevailing use of an ad-hoc number of only top-ranked features. On a wide range of data sets, our method displayed highly competitive accuracy compared to the best performance of other kinds of classification models. In addition to accuracy, our method also provides comprehensible rules to help elucidate the translation between raw data and useful knowledge. PMID- 14534179 TI - Modeling sequencing errors by combining Hidden Markov models. AB - Among the largest resources for biological sequence data is the large amount of expressed sequence tags (ESTs) available in public and proprietary databases. ESTs provide information on transcripts but for technical reasons they often contain sequencing errors. Therefore, when analyzing EST sequences computationally, such errors must be taken into account. Earlier attempts to model error prone coding regions have shown good performance in detecting and predicting these while correcting sequencing errors using codon usage frequencies. In the research presented here, we improve the detection of translation start and stop sites by integrating a more complex mRNA model with codon usage bias based error correction into one hidden Markov model (HMM), thus generalizing this error correction approach to more complex HMMs. We show that our method maintains the performance in detecting coding sequences. PMID- 14534180 TI - A suffix tree approach to the interpretation of tandem mass spectra: applications to peptides of non-specific digestion and post-translational modifications. AB - MOTIVATION: Tandem mass spectrometry combined with sequence database searching is one of the most powerful tools for protein identification. As thousands of spectra are generated by a mass spectrometer in one hour, the speed of database searching is critical, especially when searching against a large sequence database, or when the peptide is generated by some unknown or non-specific enzyme, even or when the target peptides have post-translational modifications (PTM). In practice, about 70-90% of the spectra have no match in the database. Many believe that a significant portion of them are due to peptides of non specific digestions by unknown enzymes or amino acid modifications. In another case, scientists may choose to use some non-specific enzymes such as pepsin or thermolysin for proteolysis in proteomic study, in that not all proteins are amenable to be digested by some site-specific enzymes, and furthermore many digested peptides may not fall within the rang of molecular weight suitable for mass spectrometry analysis. Interpreting mass spectra of these kinds will cost a lot of computational time of database search engines. OVERVIEW: The present study was designed to speed up the database searching process for both cases. More specifically speaking, we employed an approach combining suffix tree data structure and spectrum graph. The suffix tree is used to preprocess the protein sequence database, while the spectrum graph is used to preprocess the tandem mass spectrum. We then search the suffix tree against the spectrum graph for candidate peptides. We design an efficient algorithm to compute a matching threshold with some statistical significance level, e.g. p = 0.01, for each spectrum, and use it to select candidate peptides. Then we rank these peptides using a SEQUEST-like scoring function. The algorithms were implemented and tested on experimental data. For post-translational modifications, we allow arbitrary number of any modification to a protein. AVAILABILITY: The executable program and other supplementary materials are available online at: http://hto c.usc.edu:8000/msms/suffix/. PMID- 14534181 TI - Artificial gene networks for objective comparison of analysis algorithms. AB - MOTIVATION: Large-scale gene expression profiling generates data sets that are rich in observed features but poor in numbers of observations. The analysis of such data sets is a challenge that has been object of vigorous research. The algorithms in use for this purpose have been poorly documented and rarely compared objectively, posing a problem of uncertainty about the outcomes of the analyses. One way to objectively test such analysis algorithms is to apply them on computational gene network models for which the mechanisms are completely know. RESULTS: We present a system that generates random artificial gene networks according to well-defined topological and kinetic properties. These are used to run in silico experiments simulating real laboratory microarray experiments. Noise with controlled properties is added to the simulation results several times emulating measurement replicates, before expression ratios are calculated. AVAILABILITY: The data sets and kinetic models described here are available from http://www.vbi.vt.edu/~mendes/AGN/as biochemical dynamic models in SBML and Gepasi formats. PMID- 14534182 TI - MCMC genome rearrangement. AB - MOTIVATION: As more and more genomes have been sequenced, genomic data is rapidly accumulating. Genome-wide mutations are believed more neutral than local mutations such as substitutions, insertions and deletions, therefore phylogenetic investigations based on inversions, transpositions and inverted transpositions are less biased by the hypothesis on neutral evolution. Although efficient algorithms exist for obtaining the inversion distance of two signed permutations, there is no reliable algorithm when both inversions and transpositions are considered. Moreover, different type of mutations happen with different rates, and it is not clear how to weight them in a distance based approach. RESULTS: We introduce a Markov Chain Monte Carlo method to genome rearrangement based on a stochastic model of evolution, which can estimate the number of different evolutionary events needed to sort a signed permutation. The performance of the method was tested on simulated data, and the estimated numbers of different types of mutations were reliable. Human and Drosophila mitochondrial data were also analysed with the new method. The mixing time of the Markov Chain is short both in terms of CPU times and number of proposals. AVAILABILITY: The source code in C is available on request from the author. PMID- 14534183 TI - Gene networks inference using dynamic Bayesian networks. AB - This article deals with the identification of gene regulatory networks from experimental data using a statistical machine learning approach. A stochastic model of gene interactions capable of handling missing variables is proposed. It can be described as a dynamic Bayesian network particularly well suited to tackle the stochastic nature of gene regulation and gene expression measurement. Parameters of the model are learned through a penalized likelihood maximization implemented through an extended version of EM algorithm. Our approach is tested against experimental data relative to the S.O.S. DNA Repair network of the Escherichia coli bacterium. It appears to be able to extract the main regulations between the genes involved in this network. An added missing variable is found to model the main protein of the network. Good prediction abilities on unlearned data are observed. These first results are very promising: they show the power of the learning algorithm and the ability of the model to capture gene interactions. PMID- 14534185 TI - The shortest common supersequence problem in a microarray production setting. AB - MOTIVATION: During microarray production, several thousands of oligonucleotides (short DNA sequences) are synthesized in parallel, one nucleotide at a time. We are interested in finding the shortest possible nucleotide deposition sequence to synthesize all oligos in order to reduce production time and increase oligo quality. Thus we study the shortest common super-sequence problem of several thousand short strings over a four-letter alphabet. RESULTS: We present a statistical analysis of the basic ALPHABET-LEFTMOST approximation algorithm, and propose several practical heuristics to reduce the length of the super-sequence. Our results show that it is hard to beat ALPHABET-LEFTMOST in the microarray production setting by more than 2 characters, but these savings can improve overall oligo quality by more than four percent. AVAILABILITY: Source code in C may be obtained by contacting the author, or from http://oligos.molgen.mpg.de. PMID- 14534184 TI - Finding subtle motifs by branching from sample strings. AB - Many motif finding algorithms apply local search techniques to a set of seeds. For example, GibbsDNA (Lawrence et al. 1993, Science, 262, 208-214) applies Gibbs sampling to random seeds, and MEME (Bailey and Elkan, 1994, Proceedings of the Second International Conference on Intelligent Systems for Molecular Biology (ISMB-94), 28-36) applies the EM algorithm to selected sample strings, i.e. substrings of the sample. In the case of subtle motifs, recent benchmarking efforts show that both random seeds and selected sample strings may never get close to the globally optimal motif. We propose a new approach which searches motif space by branching from sample strings, and implement this idea in both pattern-based and profile-based settings. Our PatternBranching and ProfileBranching algorithms achieve favorable results relative to other motif finding algorithms. AVAILABILITY: http://www cse.ucsd.edu/groups/bioinformatics/software.html PMID- 14534187 TI - A description of dynamical graphs associated to elementary regulatory circuits. AB - The biological and dynamical importance of feedback circuits in regulatory graphs has often been emphasized. The work presented here aims at completely describing the dynamics of isolated elementary regulatory circuits. Our analytical approach is based on a discrete formal framework, built upon the logical approach of R. Thomas. Given a regulatory circuit, we show that the structure of synchronous and asynchronous dynamical graphs depends only on the length of the circuit (number of genes) and on its sign (which depends on the parity of the number of negative interactions). This work constitutes a first step towards the analytical characterisation of discrete dynamical graphs for more complex regulatory networks in terms of contributions corresponding to their embedded elementary circuits. PMID- 14534186 TI - Reconstructing tumor genome architectures. AB - Although cancer progression is often associated with genome rearrangements, little is known about the detailed genomic architecture of tumor genomes. The attempt to reconstruct the genomic organization of a tumor genome recently resulted in the development of the End Sequence Profiling (ESP) technique, and the application of this technique to human MCF7 tumor cells. We formulate the ESP Genome Reconstruction Problem, and develop an algorithm to solve this problem in the case of sparse ESP data. We apply our algorithm to analyze human MCF7 tumor cells, and obtain the first reconstruction of the putative architecture of human MCF7 tumor genome. Our results assist in the ongoing ESP analysis of MCF7 tumors by suggesting additional ESP experiments for the completion of a reliable reconstruction of the MCF7 tumor genome, and by focusing BAC re-sequencing efforts. PMID- 14534188 TI - Fast feature selection using a simple estimation of distribution algorithm: a case study on splice site prediction. AB - MOTIVATION: Feature subset selection is an important preprocessing step for classification. In biology, where structures or processes are described by a large number of features, the elimination of irrelevant and redundant information in a reasonable amount of time has a number of advantages. It enables the classification system to achieve good or even better solutions with a restricted subset of features, allows for a faster classification, and it helps the human expert focus on a relevant subset of features, hence providing useful biological knowledge. RESULTS: We present a heuristic method based on Estimation of Distribution Algorithms to select relevant subsets of features for splice site prediction in Arabidopsis thaliana. We show that this method performs a fast detection of relevant feature subsets using the technique of constrained feature subsets. Compared to the traditional greedy methods the gain in speed can be up to one order of magnitude, with results being comparable or even better than the greedy methods. This makes it a very practical solution for classification tasks that can be solved using a relatively small amount of discriminative features (or feature dependencies), but where the initial set of potential discriminative features is rather large. PMID- 14534189 TI - Divide-and-conquer multiple alignment with segment-based constraints. AB - A large number of methods for multiple sequence alignment are currently available. Recent benchmarking tests demonstrated that strengths and drawbacks of these methods differ substantially. Global strategies can be outperformed by approaches based on local similarities and vice versa, depending on the characteristics of the input sequences. In recent years, mixed approaches that include both global and local features have shown promising results. Herein, we introduce a new algorithm for multiple sequence alignment that integrates the global divide-and-conquer approach with the local segment-based approach, thereby combining the strengths of those two strategies. PMID- 14534190 TI - Biclustering microarray data by Gibbs sampling. AB - MOTIVATION: Gibbs sampling has become a method of choice for the discovery of noisy patterns, known as motifs, in DNA and protein sequences. Because handling noise in microarray data presents similar challenges, we have adapted this strategy to the biclustering of discretized microarray data. RESULTS: In contrast with standard clustering that reveals genes that behave similarly over all the conditions, biclustering groups genes over only a subset of conditions for which those genes have a sharp probability distribution. We have opted for a simple probabilistic model of the biclusters because it has the key advantage of providing a transparent probabilistic interpretation of the biclusters in the form of an easily interpretable fingerprint. Furthermore, Gibbs sampling does not suffer from the problem of local minima that often characterizes Expectation Maximization. We demonstrate the effectiveness of our approach on two synthetic data sets as well as a data set from leukemia patients. PMID- 14534191 TI - Finding optimal degenerate patterns in DNA sequences. AB - MOTIVATION: The problem of finding transcription factor binding sites in the upstream regions of given genes is algorithmically an interesting and challenging problem in computational biology. A degenerate pattern over a finite alphabet Sigma is a sequence of subsets of Sigma. A string over IUPAC nucleic acid codes is also a degenerate pattern over Sigma = {A, C, G, T}, and is used as one of the major patterns modeling transcription factor binding sites in the upstream regions of genes. However, it is known that the problem of finding a degenerate pattern consistent with both positive and negative string sets is in general NP complete. Our aim is to devise a heuristic algorithm to find a degenerate pattern which is optimal for positive and negative string sets w.r.t. a given score function. RESULTS: We have proposed an enumerative algorithm called SUPERPOSITION for finding optimal degenerate patterns with a pruning technique, which works with most all reasonable score functions. The performance score of the algorithm has been compared with those of other popular motif-finding algorithms YMF, MEME and AlignACE on various sets of co-regulated genes of yeast. In the computational experiment, SUPERPOSITION has outperformed the others on several gene sets. AVAILABILITY: The python script SUPERPOSITION is available at http://www.math.kyushu-u.ac.jp/~om/softwares.html PMID- 14534192 TI - Gene prediction with a hidden Markov model and a new intron submodel. AB - MOTIVATION: The problem of finding the genes in eukaryotic DNA sequences by computational methods is still not satisfactorily solved. Gene finding programs have achieved relatively high accuracy on short genomic sequences but do not perform well on longer sequences with an unknown number of genes in them. Here existing programs tend to predict many false exons. RESULTS: We have developed a new program, AUGUSTUS, for the ab initio prediction of protein coding genes in eukaryotic genomes. The program is based on a Hidden Markov Model and integrates a number of known methods and submodels. It employs a new way of modeling intron lengths. We use a new donor splice site model, a new model for a short region directly upstream of the donor splice site model that takes the reading frame into account and apply a method that allows better GC-content dependent parameter estimation. AUGUSTUS predicts on longer sequences far more human and drosophila genes accurately than the ab initio gene prediction programs we compared it with, while at the same time being more specific. AVAILABILITY: A web interface for AUGUSTUS and the executable program are located at http://augustus.gobics.de. PMID- 14534194 TI - Estimating gene networks from gene expression data by combining Bayesian network model with promoter element detection. AB - We present a statistical method for estimating gene networks and detecting promoter elements simultaneously. When estimating a network from gene expression data alone, a common problem is that the number of microarrays is limited compared to the number of variables in the network model, making accurate estimation a difficult task. Our method overcomes this problem by integrating the microarray gene expression data and the DNA sequence information into a Bayesian network model. The basic idea of our method is that, if a parent gene is a transcription factor, its children may share a consensus motif in their promoter regions of the DNA sequences. Our method detects consensus motifs based on the structure of the estimated network, then re-estimates the network using the result of the motif detection. We continue this iteration until the network becomes stable. To show the effectiveness of our method, we conducted Monte Carlo simulations and applied our method to Saccharomyces cerevisiae data as a real application. PMID- 14534196 TI - Extracting active pathways from gene expression data. AB - MOTIVATION: A promising way to make sense out of gene expression profiles is to relate them to the activity of metabolic and signalling pathways. Each pathway usually involves many genes, such as enzymes, which can themselves participate in many pathways. The set of all known pathways can therefore be represented by a complex network of genes. Searching for regularities in the set of gene expression profiles with respect to the topology of this gene network is a way to automatically extract active pathways and their associated patterns of activity. METHOD: We present a method to perform this task, which consists in encoding both the gene network and the set of profiles into two kernel functions, and performing a regularized form of canonical correlation analysis between the two kernels. RESULTS: When applied to publicly available expression data the method is able to extract biologically relevant expression patterns, as well as pathways with related activity. PMID- 14534198 TI - Flexible structure alignment by chaining aligned fragment pairs allowing twists. AB - MOTIVATION: Protein structures are flexible and undergo structural rearrangements as part of their function, and yet most existing protein structure comparison methods treat them as rigid bodies, which may lead to incorrect alignment. RESULTS: We have developed the Flexible structure AlignmenT by Chaining AFPs (Aligned Fragment Pairs) with Twists (FATCAT), a new method for structural alignment of proteins. The FATCAT approach simultaneously addresses the two major goals of flexible structure alignment; optimizing the alignment and minimizing the number of rigid-body movements (twists) around pivot points (hinges) introduced in the reference protein. In contrast, currently existing flexible structure alignment programs treat the hinge detection as a post-process of a standard rigid body alignment. We illustrate the advantages of the FATCAT approach by several examples of comparison between proteins known to adopt different conformations, where the FATCAT algorithm achieves more accurate structure alignments than current methods, while at the same time introducing fewer hinges. PMID- 14534199 TI - Association between chloroplast DNA and mitochondrial DNA haplotypes in Prunus spinosa L. (Rosaceae) populations across Europe. AB - Chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) were studied in 24 populations of Prunus spinosa sampled across Europe. The cpDNA and mtDNA fragments were amplified using universal primers and subsequently digested with restriction enzymes to obtain the polymorphisms. Combinations of all the polymorphisms resulted in 33 cpDNA haplotypes and two mtDNA haplotypes. Strict association between the cpDNA haplotypes and the mtDNA haplotypes was detected in most cases, indicating conjoint inheritance of the two genomes. The most frequent and abundant cpDNA haplotype (C20; frequency, 51 %) is always associated with the more frequent and abundant mtDNA haplotype (M1; frequency, 84 %). All but two of the cpDNA haplotypes associated with the less frequent mtDNA haplotype (M2) are private haplotypes. These private haplotypes are phylogenetically related but geographically unrelated. They form a separate cluster on the minimum-length spanning tree. PMID- 14534200 TI - Photosynthetic responses of the tropical spiny shrub Lycium nodosum (Solanaceae) to drought, soil salinity and saline spray. AB - Water relations and photosynthetic characteristics of plants of Lycium nodosum grown under increasing water deficit (WD), saline spray (SS) or saline irrigation (SI) were studied. Plants of this perennial, deciduous shrub growing in the coastal thorn scrubs of Venezuela show succulent leaves which persist for approx. 1 month after the beginning of the dry season; leaf succulence is higher in populations closer to the sea. These observations suggested that L. nodosum is tolerant both to WD and salinity. In the glasshouse, WD caused a marked decrease in the xylem water potential (psi), leaf osmotic potential (psi(s)) and relative water content (RWC) after 21 d; additionally, photosynthetic rate (A), carboxylation efficiency (CE) and stomatal conductance (gs) decreased by more than 90 %. In contrast, in plants treated for 21 d with a foliar spray with 35 per thousand NaCl or irrigation with a 10 % NaCl solution, psi and RWC remained nearly constant, while psi(s) decreased by 30 %, and A, CE and gs decreased by more than 80 %. An osmotic adjustment of 0.60 (SS) and 0.94 MPa (SI) was measured. Relative stomatal and mesophyll limitations to A increased with both WD and SS, but were not determined for SI-treated plants. No evidence of chronic photoinhibition due to any treatment was observed, since maximum quantum yield of PSII, Fv/Fm, did not change with either drought in the field or water or salinity stress in the glasshouse. Nevertheless, WD and SI treatments caused a decrease in the photochemical (qP) and an increase in the non-photochemical (qN) quenching coefficients relative to controls; qN was unaffected by the SS treatment. The occurrence of co-limitation of A by stomatal and non-stomatal factors in plants of L. nodosum may be associated with the extended leaf duration under water or saline stress. Additionally, osmotic adjustment may partly explain the relative maintenance of A and gs in the SS and SI treatments and the tolerance to salinity of plants of this species in coastal habitats. PMID- 14534201 TI - Roots of Pisum sativum L. exhibit hydrotropism in response to a water potential gradient in vermiculite. AB - In the present study, root hydrotropism in an agravitropic mutant of Pisum sativum L. grown in vermiculite with a steep water potential gradient was examined. When wet and dry vermiculite were placed side by side, water diffused from the wet (-0.04 MPa) to the dry (-1.2 MPa) and a steep water potential gradient became apparent in the dry vermiculite close to the boundary between the two. The extent and location of the gradient remained stable between the fourth and sixth day after filling a box with vermiculite, and the steepest gradient (approx. 0.02 MPa mm-1) was found in the initially dry vermiculite between 60 and 80 mm from the boundary. When seedlings with 25-35 mm long roots were planted in the initially dry vermiculite near where the gradient had been established, each of the main roots elongated toward the wet vermiculite, i.e. toward the high water potential. Control roots elongated without curvature in both the wet and the dry vermiculite, in which no water potential gradient was detectable. These results show that pea roots respond to the water potential gradient around them and elongate towards the higher water potential. Therefore, positive hydrotropism occurs in vermiculite just as it does in air. Hydrotropism in soil may be significant when a steep water potential gradient is apparent, such as when drip irrigation is applied. PMID- 14534202 TI - Floral structure, breeding system and fruit-set in the threatened sub-shrub Tetratheca juncea Smith (Tremandraceae). AB - Tetratheca juncea Smith (Tremandraceae) has undergone a range contraction of approx. 50 km in the last 100 years and is now listed as a vulnerable sub-shrub restricted to the central and north coast regions of New South Wales, Australia. There are approx. 250 populations in a 110 km north-south distribution and populations are usually small with fewer than 50 plants/clumps. The reproductive ecology of the species was studied to determine why seed-set is reportedly rare. Flowers are bisexual, odourless and nectarless. Flowers are presented dependentally and there are eight stamens recurved around the pistil. Anthers are poricidal, contain viable pollen and basally contain a deep-red tapetal fluid that is slightly oily. Thus flowers are presented for buzz pollinators, although none were observed at flowers during our study. The species was found to be facultatively xenogamous with only one in 50 glasshouse flowers setting seed autogamously, i.e. without pollinator assistance. Field studies revealed fertile fruit in 24 populations but production varied significantly across sites from exceedingly low (0.6 fruits per plant clump) to low (17 fruits per plant clump). Fruit-set ranged from 0 to 65 %, suggesting that pollen vectors exist or that autogamy levels in the field are variable and higher than glasshouse results. Fruit production did not vary with population size, although in three of the five populations in the south-west region more than twice as much fruit was produced as in populations elsewhere. A moderately strong relationship between foliage volume and fruit : flower ratios suggests that bigger plants may be more attractive than smaller plants to pollinators. A review of Tetratheca pollination ecology revealed that several species are poorly fecund and pollinators are rare. The habitat requirements for Tetratheca, a genus of many rare and threatened species, is discussed. PMID- 14534204 TI - Immigrant status and smoking. PMID- 14534206 TI - Mandatory HIV testing in prisons. PMID- 14534207 TI - Eliminating health inequalities. PMID- 14534208 TI - Achieving health equity: an incremental journey. PMID- 14534209 TI - The Veterans Health administration's promotion of health equity for racial and ethnic minorities. PMID- 14534210 TI - Addressing racial and ethnic health disparities. PMID- 14534211 TI - Images in public health. PMID- 14534212 TI - Restrictions on undocumented immigrants' access to health services: the public health implications of welfare reform. AB - The Personal Responsibility and Work Opportunity Reconciliation Act of 1996 greatly restricts the provision of many federal, state, and local public services to undocumented immigrants. These restrictions have prompted intense debates about the provision of free and discounted primary and preventive health care services and have placed significant burdens on institutions that serve large undocumented immigrant populations. Intended to serve as a tool for reducing illegal immigration and protecting public resources, federal restrictions on undocumented immigrants' access to publicly financed health services unduly burden health care providers and threaten the public's health. These deleterious effects warrant the public health community's support of strategies designed to sustain provision of health services irrespective of immigration status. PMID- 14534213 TI - Tribal benefits counseling program: expanding health care opportunities for tribal members. AB - American Indian tribal clinics hired benefits counselors to increase the number of patients with public and private insurance coverage, expand the range of health care options available to tribal members, and increase third-party revenues for tribal clinics. Benefits counselors received intensive training, technical assistance, and evaluation over a 2-year period. Six tribal clinics participated in the full training program, including follow-up, process evaluation, and outcomes reporting. Participating tribal sites experienced a 78% increase in Medicaid enrollment among pregnant women and children, compared with a 26% enrollment increase statewide during the same period. Trained benefits counselors on-site at tribal clinics can substantially increase third-party insurance coverage among patients. PMID- 14534214 TI - A no-name tuberculosis tracking system. AB - Foreign-born persons from countries where tuberculosis (TB) is endemic make up a significant percentage of poultry industry workers in Delaware, a leading poultry producing state. Many of these workers enter the United States without documentation and assume multiple identities, making it difficult for public health staff to investigate TB contacts who work in the poultry plants. The Sussex County Health Unit of the Delaware Division of Public Health developed a no-name TB tracking system to facilitate identification and treatment of poultry plant workers with TB infection and disease in a high-risk population whose members assume one or more aliases. Completion rates for treatment of latent TB infection in this group increased from 48% to 64% 2 years after the program's implementation. PMID- 14534215 TI - A global network for early warning and response to infectious diseases and bioterrorism: applied epidemiology and training programs, 2001. AB - In many ministries of health, applied epidemiology and training programs (AETPs) are responsible for detecting and responding to acute health events, including bioterrorism. In November 2001, we assessed the bioterrorism response capacity of 29 AETPs; 17 (59%) responded. Fifteen countries (88%) had bioterrorism response plans; in 6 (40%), AETPs took the lead in preparation and in 6 (40%) they assisted. Between September 11 and November 29, 2001, 12 AETPs (71%) responded to a total of 3024 bioterrorism-related phone calls. Six programs (35%) responded to suspected bioterrorism events. AETPs play an important role in bioterrorism surveillance and response. Support for this global network by various health agencies is beneficial for all developed and developing countries. PMID- 14534216 TI - Shame on you, uncle! PMID- 14534217 TI - Medical care for interned enemy aliens: a role for the US Public Health Service in World War II. AB - During World War II, the US Public Health Service (USPHS) administered health care to 19 000 enemy aliens and Axis merchant seamen interned by the Justice Department through its branch, the Immigration and Naturalization Service (INS). The Geneva Prisoners of War Convention of 1929, which the United States applied to civilian internees, provided guidelines for belligerent nations regarding humanitarian treatment of prisoners of war, including for their health. The INS forged an agreement with the USPHS to meet these guidelines for the German, Italian, and Japanese internees and, in some cases, their families. Chronic shortages and crowded camps continuously challenged USPHS administrators. Nevertheless, the USPHS offered universal access to care and provided treatment often exceeding care received by many American citizens. PMID- 14534219 TI - Parents, practitioners, and researchers: community-based participatory research with early head start. AB - Community-based participatory research (CBPR) is an approach to research and evaluation that is receiving increased attention in the field of public health. Our report discusses the application of this approach to research and evaluation with an Early Head Start (EHS) program in Pittsburgh, Pa. Our primary purpose is to illustrate the key elements that contributed to effective collaboration among researchers, EHS practitioners, and parents of EHS children in the conduct of the study. The focus is not on research findings but on research process. Our goal is to make the practices of CBPR visible and explicit so they can be analyzed, further developed, and effectively applied to a range of public health issues in a diversity of community contexts. PMID- 14534218 TI - Race/ethnicity, gender, and monitoring socioeconomic gradients in health: a comparison of area-based socioeconomic measures--the public health disparities geocoding project. AB - Use of multilevel frameworks and area-based socioeconomic measures (ABSMs) for public health monitoring can potentially overcome the absence of socioeconomic data in most US public health surveillance systems. To assess whether ABSMs can meaningfully be used for diverse race/ethnicity-gender groups, we geocoded and linked public health surveillance data from Massachusetts and Rhode Island to 1990 block group, tract, and zip code ABSMs. Outcomes comprised death, birth, cancer incidence, tuberculosis, sexually transmitted infections, childhood lead poisoning, and nonfatal weapons-related injuries. Among White, Black, and Hispanic women and men, measures of economic deprivation (e.g., percentage below poverty) were most sensitive to expected socioeconomic gradients in health, with the most consistent results and maximal geocoding linkage evident for tract-level analyses. PMID- 14534220 TI - Eligibility for government insurance if immigrant provisions of welfare reform are repealed. PMID- 14534221 TI - Racial disparities in alcohol use: comparison of 2 American Indian reservation populations with national data. PMID- 14534222 TI - Hospital policy and practice regarding the collection of data on race, ethnicity, and birthplace. PMID- 14534223 TI - Racial/ethnic variations in physician recommendations for cardiac revascularization. AB - OBJECTIVES: We sought to examine whether physician recommendations for cardiac revascularization vary according to patient race. METHODS: We studied patients scheduled for coronary angiography at 2 hospitals, one public and one private, between November 1997 and June 1999. Cardiologists were interviewed regarding their recommendations for cardiac resvacularization. RESULTS: African American patients were less likely than Whites to be recommended for revascularization at the public hospital (adjusted odds ratio [OR] = 0.31; 95% confidence interval [CI] = 0.12, 0.77) but not at the private hospital (adjusted OR = 1.69; 95% CI = 0.69, 4.14). CONCLUSIONS: Physician recommendations for cardiac revascularization vary by patient race. Further studies are needed to examine physician bias as a factor in racial disparities in cardiac care and outcomes. PMID- 14534225 TI - Tailored interventions to increase influenza vaccination in neighborhood health centers serving the disadvantaged. AB - OBJECTIVES: We designed and evaluated interventions to increase adult immunizations within inner-city health centers. METHODS: Interventions included reminders, standing orders, and walk-in "flu shot clinics." Patients were surveyed and records evaluated. RESULTS: Records from 1 center showed that immunization rates increased from 24% to 30% (P <.001) for patients aged 50 to 64 years and from 45% to 53% for patients aged 65 years and older (P <.001). Self reported vaccination rates did not increase. In logistic regression analyses, the strongest predictor of vaccination among patients aged 50 to 64 years was the belief that unvaccinated persons will contract influenza (odds ratio [OR] = 5.4; 95% confidence interval [CI] = 2.4, 12.0). Among patients aged 65 years and older, the strongest predictor of vaccination was the belief that friends/relatives thought that they should be vaccinated (OR = 9.7; 95% CI = 4.2, 22.3). CONCLUSIONS: Tailored interventions can improve immunization rates at inner-city health centers. PMID- 14534224 TI - Health behaviors and quality of care among Latinos with diabetes in managed care. AB - OBJECTIVES: We evaluated whether ethnicity and language are associated with diabetes care for Latinos in managed care. METHODS: Using data from 4685 individuals in the Translating Research Into Action for Diabetes (TRIAD) Study, a multicenter study of diabetes care in managed care, we constructed multivariate regression models to compare health behaviors, processes of care, and intermediate outcomes for Whites and English- and Spanish-speaking Latinos. RESULTS: Latinos had lower rates of self-monitoring of blood glucose and worse glycemic control than did Whites, higher rates of foot self-care and dilated-eye examinations, and comparable rates of other processes and intermediate outcomes of care. CONCLUSIONS: Although self-management and quality of care are comparable for Latinos and Whites with diabetes, important ethnic disparities persist in the managed care settings studied. PMID- 14534226 TI - Determinants of mortality following a diagnosis of prostate cancer in Veterans Affairs and private sector health care systems. AB - OBJECTIVES: We compared patterns of mortality among men with prostate cancer at 2 Department of Veterans Affairs (VA) and 2 private-sector hospitals in the Chicago area. METHODS: Mortality rates for 864 cases diagnosed between 1986 and 1990 were estimated using Cox proportional hazards models that incorporated age; income; cancer stage, differentiation, and treatments; and baseline comorbidity. RESULTS: Race tended to associate with all-cause mortality irrespective of health care setting (Blacks vs Whites: hazard rate ratio [HRR] = 1.68 [95% confidence interval (CI) = 1.06, 2.67]; P <.001 in the private sector; HRR = 1.50 [95% CI = 0.94, 2.38]; P =.088 in the VA). However, comorbidity determined risk in the VA, whereas age and income predicted risk in the private sector. CONCLUSIONS: Determinants of all-cause mortality in men with prostate cancer vary according to health care setting. PMID- 14534227 TI - Patient-physician relationships and racial disparities in the quality of health care. AB - OBJECTIVES: This study explored whether racial differences in patient-physician relationships contribute to disparities in the quality of health care. METHODS: We analyzed data from The Commonwealth Fund's 2001 Health Care Quality Survey to determine whether racial differences in patients' satisfaction with health care and use of basic health services were explained by differences in quality of patient-physician interactions, physicians' cultural sensitivity, or patient physician racial concordance. RESULTS: Both satisfaction with and use of health services were lower for Hispanics and Asians than for Blacks and Whites. Racial differences in the quality of patient-physician interactions helped explain the observed disparities in satisfaction, but not in the use of health services. CONCLUSIONS: Barriers in the patient-physician relationship contribute to racial disparities in the experience of health care. PMID- 14534228 TI - Research expectations among African American church leaders in the PRAISE! project: a randomized trial guided by community-based participatory research. AB - OBJECTIVES: This study sought to examine the expectations and satisfaction of pastors and lay leaders regarding a research partnership in a randomized trial guided by community-based participatory research (CBPR) methods. METHODS: Telephone and self-administered print surveys were administered to 78 pastors and lay leaders. In-depth interviews were conducted with 4 pastors after study completion. RESULTS: The combined survey response rate was 65%. Research expectations included honest and frequent communication, sensitivity to the church environment, interaction as partners, and results provided to the churches. Satisfaction with the research partnership was high, but so was concern about the need for all research teams to establish trust with church partners. CONCLUSIONS: Pastors and lay leaders have high expectations regarding university obligations in research partnerships. An intervention study based on CBPR methods was able to meet most of these expectations. PMID- 14534229 TI - Understanding racial disparities in HIV using data from the veterans aging cohort 3-site study and VA administrative data. AB - OBJECTIVES: We identified race-associated differences in survival among HIV positive US veterans to examine possible etiologies for these differences. METHODS: We used national administrative data to compare survival by race and used data from the Veterans Aging Cohort 3-Site Study (VACS 3) to compare patients' health status, clinical management, and adherence to medication by race. RESULTS: Nationally, minority veterans had higher mortality rates than did white veterans with HIV. Minority veterans had poorer health than white veterans with HIV. No significant differences were found in clinical management or adherence. CONCLUSIONS: HIV-positive minority veterans experience poorer survival than white veterans. This difference may derive from differences in comorbidities and in the severity of illness of HIV-related disease. PMID- 14534230 TI - Agreement between administrative data and patients' self-reports of race/ethnicity. AB - OBJECTIVES: We examined agreement of administrative data with self-reported race/ethnicity and identified correlates of agreement. METHODS: We used Veterans Affairs administrative data and VA 1999 Large Health survey race/ethnicity data. RESULTS: Relatively low rates of agreement (approximately 60%) between data sources were largely the result of administrative data from patients whose race/ethnicity was unknown, with least agreement for Native American, Asian, and Pacific Islander patients. After exclusion of patients with missing race/ethnicity, agreement improved except for Native Americans. Agreement did not increase substantially after inclusion of data from individuals indicating multiple race/ethnicities. Patients for whom there was better agreement between data sources tended to be less educated, non-solitary living, younger, and White; to have sufficient food; and to use more inpatient Department of Veterans Affairs (VA) care. CONCLUSIONS: Better reporting of race/ethnicity data will improve agreement between data sources. Previous studies using VA administrative data may have underestimated racial disparities. PMID- 14534231 TI - Health status, health insurance, and health care utilization patterns of immigrant Black men. AB - OBJECTIVES: This study sought to describe the health status, health insurance, and health care utilization patterns of the growing population of immigrant Black men. METHODS: We used data from the 1997-2000 National Health Interview Survey to examine and then compare health variables of foreign-born Black men with those of US-born Black and White men. Logistic regression analyses were used to examine health outcomes. RESULTS: Foreign-born Black men were in better overall health than their US-born Black counterparts and were much less likely than either US born Black or White men to report adverse health behaviors. Despite these health advantages, foreign-born Black men were more likely than either US-born Black or White men to be uninsured. CONCLUSIONS: In the long term, immigrant Black men who are in poor health may be adversely affected by lack of health care coverage. PMID- 14534232 TI - Occupational risk factors for selected cancers among African American and White men in the United States. AB - OBJECTIVES: This study examined occupational risks for non-Hodgkin's lymphoma, Hodgkin's disease, and soft-tissue sarcoma among African American and White men. METHODS: Race-specific multivariate logistic regression analyses were conducted using data from a large US population-based case-control study. RESULTS: Significant occupational risks were limited to African Americans; chromium was associated with non-Hodgkin's lymphoma (odds ratio [OR] = 3.9, 95% confidence interval [CI] = 1.2, 12.9) and wood dust was associated with Hodgkin's disease (OR = 4.6, 95% CI = 1.6, 13.3) and soft-tissue sarcoma (OR = 3.7, 95% CI = 1.6, 8.6). CONCLUSIONS: Race-specific occupational risk factors for cancer were evident only among African American men. This may reflect racial disparities in levels of exposure to occupational carcinogens. PMID- 14534234 TI - Human alpha4beta2 acetylcholine receptors formed from linked subunits. AB - We prepared concatamers of alpha4 and beta2 subunits for human nicotinic acetylcholine receptors (AChRs), in which the C terminus of alpha4 was linked to the N terminus of beta2, or vice versa, via a tripeptide sequence repeated 6 or 12 times, and expressed them in Xenopus oocytes. Linkage did not substantially alter channel amplitude or channel open-duration. Linkage at the C terminus of alpha4 prevented AChR potentiation by 17-beta-estradiol by disruption of its binding site. Assembly of AChRs from concatamers was less efficient, but function was much more efficient than that of unlinked subunits. With both linked and free subunits, greater ACh-induced currents per surface AChR were observed with the (alpha4)3(beta2)2 stoichiometry than with the (alpha4)2(beta2)3 stoichiometry. The (alpha4)3(beta2)2 stoichiometry exhibited much lower ACh sensitivity. When concatamers were expressed alone, dipentameric AChRs were formed in which the (alpha4)2(beta2)3 pentamer was linked to the (alpha4)3(beta2)2 pentamer. Dipentamers were selectively expressed on the cell surface, whereas most monopentamers with dangling subunits were retained intracellularly. Coexpression of concatamers with monomeric beta2, beta4, or alpha4 subunits resulted in monopentamers, the stoichiometry of which was determined by the free subunit added. Linkage between the C terminus of beta2 and the N terminus of alpha4 favored formation of ACh-binding sites within the concatamer, whereas linkage between the C terminus of alpha4 and the N terminus of beta2 favored formation of ACh-binding sites between concatamers. These protein-engineering studies provide insight into the structure and function of alpha4beta2 AChRs, emphasizing the functional differences between alpha4beta2 AChRs of different stoichiometries. PMID- 14534233 TI - Stage at diagnosis and survival in a multiethnic cohort of prostate cancer patients. AB - OBJECTIVES: We evaluated the effects of socioeconomic status and comorbidity on stage of disease and survival among 1509 population-based prostate cancer patients. METHODS: We applied logistic regression and Cox proportional hazards regression to data from Whites, African Americans, and Asian Americans who were diagnosed from 1987 to 1991. RESULTS: Patients with existing comorbid conditions were less likely than those without these conditions to be diagnosed with advanced cancer. Compared with Whites, African Americans (odds ratio [OR] = 1.5; 95% confidence interval [CI] = 1.1, 2.2) and foreign-born Asian Americans (OR = 1.6; 95% CI = 1.0, 2.4) were more likely to be diagnosed with advanced cancer. Among men with localized disease, prostate cancer death rates were higher for African Americans than for Whites (death rate ratio = 2.3; 95% CI = 1.2, 4.7). CONCLUSIONS: These findings support the need for further investigation of factors that affect access to and use of health care among African Americans and Asian Americans. PMID- 14534235 TI - Improvement in fear memory by histamine-elicited ERK2 activation in hippocampal CA3 cells. AB - Consolidation of associative memories appears to require extracellular signal related kinase2 (ERK2) activation, which is modulated by several factors, including neurotransmitter receptor stimulation. Here we show that in vitro stimulation of either H2 or H3 histaminergic receptors activates ERK2 in hippocampal CA3 pyramidal cells. In behaving animals, bilateral posttraining injections into the dorsal hippocampus of histamine H2 or H3 receptor agonists improve memory consolidation after contextual fear conditioning. Local administration of U0126, a selective inhibitor of ERK kinase, prevents memory improvements exerted by the agonists, without causing any behavioral effect per se. This is the first evidence of a positive correlation between ERK phosphorylation and memory improvement. Moreover, we demonstrate that the brain histaminergic system regulates hippocampal ERK cascade. Finally, our data indicate that early ERK2 hippocampal activation is not required for the expression of long-term fear memories. PMID- 14534236 TI - Functional mapping and Ca2+ regulation of nicotinic acetylcholine receptor channels in rat hippocampal CA1 neurons. AB - Diverse subtypes of nicotinic acetylcholine receptors (nAChRs), including fast desensitizing alpha7-containing receptors thought to be Ca2+-permeable, are expressed in the CNS, where they appear to regulate cognitive processing and synaptic plasticity. To understand the physiological role of nAChRs in regulating neuronal excitability, it is important to know the distribution of functional receptors along the surface of neurons, whether they can increase [Ca2+]i, and/or are regulated by Ca2+. We mapped the distribution of receptors on the membrane of rat hippocampal CA1 stratum radiatum interneurons and pyramidal cells in acute slices by recording nAChR-mediated currents elicited by local UV laser-based photolysis of caged carbachol in patch-clamped neurons. The local application (approximately 7 microm patches) allowed mapping of functional nAChRs along the soma and dendritic tree, whereas the fast uncaging minimized the effects of desensitization of alpha7-containing nAChRs and allowed us to measure the kinetics of responses. The alpha7-containing nAChRs were the predominant subtype on interneurons, and were located primarily at perisomatic sites (<70 microm from the soma; in contrast to the more uniform distribution of glutamate receptors); no currents were detectable on pyramidal neurons. The activation of nAChRs increased [Ca2+]i, indicating that these native receptors in acute slices are significantly Ca2+-permeable, consistent with previous observations made with recombinant receptors. In addition, they exhibited strong desensitization, the rate of recovery from which was controlled by [Ca2+]i. Our results demonstrate the strategic location and Ca2+ regulation of alpha7-containing nAChRs, which may contribute to understanding their involvement in hippocampal plasticity. PMID- 14534237 TI - Quantifying generalization from trial-by-trial behavior of adaptive systems that learn with basis functions: theory and experiments in human motor control. AB - During reaching movements, the brain's internal models map desired limb motion into predicted forces. When the forces in the task change, these models adapt. Adaptation is guided by generalization: errors in one movement influence prediction in other types of movement. If the mapping is accomplished with population coding, combining basis elements that encode different regions of movement space, then generalization can reveal the encoding of the basis elements. We present a theory that relates encoding to generalization using trial by-trial changes in behavior during adaptation. We consider adaptation during reaching movements in various velocity-dependent force fields and quantify how errors generalize across direction. We find that the measurement of error is critical to the theory. A typical assumption in motor control is that error is the difference between a current trajectory and a desired trajectory (DJ) that does not change during adaptation. Under this assumption, in all force fields that we examined, including one in which force randomly changes from trial to trial, we found a bimodal generalization pattern, perhaps reflecting basis elements that encode direction bimodally. If the DJ was allowed to vary, bimodality was reduced or eliminated, but the generalization function accounted for nearly twice as much variance. We suggest, therefore, that basis elements representing the internal model of dynamics are sensitive to limb velocity with bimodal tuning; however, it is also possible that during adaptation the error metric itself adapts, which affects the implied shape of the basis elements. PMID- 14534238 TI - Mechanisms of mitochondria-neurofilament interactions. AB - Mitochondria are localized to regions of the cell where ATP consumption is high and are dispersed according to changes in local energy needs. In addition to motion directed by molecular motors, mitochondrial distribution in neuronal cells appears to depend on the docking of mitochondria to microtubules and neurofilaments. We examined interactions between mitochondria and neurofilaments using fluorescence microscopy, dynamic light scattering, atomic force microscopy, and sedimentation assays. Mitochondria-neurofilament interactions depend on mitochondrial membrane potential, as revealed by staining with a membrane potential sensitive dye (JC-1) in the presence of substrates/ADP or uncouplers (valinomycin/carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone) and are affected by the phosphorylation status of neurofilaments and neurofilament sidearms. Antibodies against the neurofilament heavy subunit disrupt binding between mitochondria and neurofilaments, and isolated neurofilament sidearms alone interact with mitochondria, suggesting that they mediate the interactions between the two structures. These data suggest that specific and regulated mitochondrial-neurofilament interactions occur in situ and may contribute to the dynamic distribution of these organelles within the cytoplasm of neurons. PMID- 14534239 TI - Overexpression of a hyperpolarization-activated cation current (Ih) channel gene modifies the firing activity of identified motor neurons in a small neural network. AB - The hyperpolarization-activated cation current (Ih) is widely distributed in excitable cells. Ih plays important roles in regulation of cellular excitability, rhythmic activity, and synaptic function. We previously showed that, in pyloric dilator (PD) neurons of the stomatogastric ganglion (STG) of spiny lobsters, Ih can be endogenously upregulated to compensate for artificial overexpression of the Shal transient potassium channel; this maintains normal firing properties of the neuron despite large increases in potassium current. To further explore the function of Ih in the pyloric network, we injected cRNA of PAIH, a lobster gene that encodes Ih, into rhythmically active PD neurons. Overexpression of PAIH produced a fourfold increase in Ih, although with somewhat different biophysical properties than the endogenous current. Compared with the endogenous Ih, the voltage for half-maximal activation of the PAIH-evoked current was depolarized by 10 mV, and its activation kinetics were significantly faster. This increase in Ih did not affect the expression of IA or other outward currents. Instead, it significantly altered the firing properties of the PD neurons. Increased Ih depolarized the minimum membrane potential of the cell, reduced the oscillation amplitude, decreased the time to the first spike, and increased the duty cycle and number of action potentials per burst. We used both dynamic-clamp experiments, injecting the modeled PAIH currents into PD cells in a functioning STG, and a theoretical model of a two-cell network to demonstrate that the increased Ih was sufficient to cause the observed changes in the PD activity. PMID- 14534240 TI - The spinal interneurons and properties of glutamatergic synapses in a primitive vertebrate cutaneous flexion reflex. AB - Unlike the monosynaptic "stretch" reflex, the exact neuronal pathway for a simple cutaneous reflex has not yet been defined in any vertebrate. In young frog tadpoles, we made whole-cell recordings from pairs of spinal neurons. We found direct, excitatory, glutamatergic synapses from touch-sensitive skin-sensory neurons to sensory pathway interneurons, and then from these sensory interneurons to motoneurons and premotor interneurons on the other side of the body. We conclude that the minimal pathway for this primitive reflex, in which stroking the skin on one side leads to flexion on the other side, is disynaptic. This detailed circuit information has allowed us to ask whether the properties of glutamatergic synapses during the first day of CNS development are tuned to their function in the tadpole's responses. Stroking the skin excites a few sensory neurons. These activate primarily AMPA receptors producing short, strong excitation that activates many sensory pathway interneurons but only allows temporal summation of closely synchronous inputs. In contrast, the excitation produced in contralateral neurons by the sensory pathway interneurons is weak and primarily mediated by NMDA receptors. As a result, considerable summation is required for this excitation to lead to postsynaptic neuron firing and a contralateral flexion. We conclude that from their early functioning, synapses from sensory neurons are strong and those from sensory pathway interneurons are weak. The distribution of glutamate receptors at synapses in this developing circuit is tuned so that synapses have properties suited to their roles in the whole animal's reflex responses. PMID- 14534241 TI - Conditional ablation of striatal neuronal types containing dopamine D2 receptor disturbs coordination of basal ganglia function. AB - Dopamine (DA) exerts synaptic organization of basal ganglia circuitry through a variety of neuronal populations in the striatum. We performed conditional ablation of striatal neuronal types containing DA D2 receptor (D2R) by using immunotoxin-mediated cell targeting. Mutant mice were generated that express the human interleukin-2 receptor alpha-subunit under the control of the D2R gene. Intrastriatal immunotoxin treatment of the mutants eliminated the majority of the striatopallidal medium spiny neurons and cholinergic interneurons. The elimination of these neurons caused hyperactivity of spontaneous movement and reduced motor activation in response to DA stimulation. The elimination also induced upregulation of GAD gene expression in the globus pallidus (GP) and downregulation of cytochrome oxidase activity in the subthalamic nucleus (STN), whereas it attenuated DA-induced expression of the immediate-early genes (IEGs) in the striatonigral neurons. In addition, chemical lesion of cholinergic interneurons did not alter spontaneous movement but caused a moderate enhancement in DA-induced motor activation. This enhancement of the behavior was accompanied by an increase in the IEG expression in the striatonigral neurons. These data suggest that ablation of the striatopallidal neurons causes spontaneous hyperactivity through modulation of the GP and STN activity and that the ablation leads to the reduction in DA-induced behavior at least partly through attenuation of the striatonigral activity as opposed to the influence of cholinergic cell lesion. We propose a possible model in which the striatopallidal neurons dually regulate motor behavior dependent on the state of DA transmission through coordination of the basal ganglia circuitry. PMID- 14534242 TI - Prestin and the dynamic stiffness of cochlear outer hair cells. AB - The outer hair cell (OHC) lateral wall is a unique trilaminate structure consisting of the plasma membrane, the cortical lattice, and subsurface cisternae. OHCs are capable of altering their length in response to transmembrane voltage change. This so-called electromotile response is presumed to result from conformational changes of membrane-bound protein molecules, named prestin. OHC motility is accompanied by axial stiffness changes when the membrane potential of the cell is altered. During length changes, intracellular anions (mainly Cl-) act as extrinsic voltage sensors. In this study, we inquired whether the motor proteins are responsible for the voltage-dependent axial stiffness of OHCs, and whether ACh, the neurotransmitter of efferent neurons, modulates the stiffness of the cortical lattice and/or the stiffness of the motor protein. The experiments were done on isolated guinea pig OHCs in the whole-cell voltage-clamp mode. Axial stiffness was determined by loading a fiber of known stiffness onto the apical surface of the cells. Voltage-dependent stiffness and cell motility disappeared, and the axial stiffness of the cells significantly decreased after removal of intracellular Cl-. The result suggests that the stiffness of the motor protein is a major contributor to the global axial stiffness of OHCs. ACh was found to affect both the motor protein and other lateral wall stiffness components. PMID- 14534243 TI - Altered Abeta formation and long-term potentiation in a calsenilin knock-out. AB - Calsenilin has been identified as a presenilin-binding protein, a transcription factor regulating dynorphin expression, and a beta-subunit of Kv4 channels and could, thus, be a multifunctional protein. To study these functions of calsenilin in vivo and to determine the neuroanatomical expression pattern of calsenilin, we generated mice with a disruption of the calsenilin gene by the targeted insertion of the beta-galactosidase gene. We found that calsenilin expression (as represented by beta-galactosidase activity) is very restricted but overlaps better with that of presenilins and Kv4 channels than with dynorphin, suggesting that calsenilin may regulate presenilin and Kv4 channels in brain. Abeta peptide levels are reduced in calsenilin knock-out mice, demonstrating that calsenilin affects presenilin-dependent gamma-cleavage in vivo. Furthermore, long-term potentiation (LTP) in dentate gyrus of hippocampus, in which calsenilin is strongly and selectively expressed, is enhanced in calsenilin knock-out mice. This enhancement of LTP coincides with a downregulation of the Kv4 channel dependent A-type current and can be mimicked in wild-type animals by a Kv4 channel blocker. The data presented here show that lack of calsenilin affects both Abeta formation and the A-type current. We suggest that these effects are separate events, caused by a common mechanism possibly involving protein transport. PMID- 14534244 TI - 3,4-methylenedioxymethamphetamine (ecstasy) inhibits dyskinesia expression and normalizes motor activity in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated primates. AB - Ecstasy [3,4-methylenedioxymethamphetamine (MDMA)] was shown to prolong the action of L-3,4-dihydroxyphenylalanine (L-DOPA) while suppressing dyskinesia in a single patient with Parkinson's disease (PD). The clinical basis of this effect of MDMA is unknown but may relate to its actions on either dopaminergic or serotoninergic systems in brain. In normal, drug-naive common marmosets, MDMA administration suppressed motor activity and exploratory behavior. In 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated, L-DOPA-primed common marmosets, MDMA transiently relieved motor disability but over a period of 60 min worsened motor symptoms. When given in conjunction with L-DOPA, however, MDMA markedly decreased dyskinesia by reducing chorea and to a lesser extent dystonia and decreased locomotor activity to the level observed in normal animals. MDMA similarly alleviated dyskinesia induced by the selective dopamine D2/3 agonist pramipexole. The actions of MDMA appeared to be mediated through 5-HT mechanisms because its effects were fully blocked by the selective serotonin reuptake inhibitor fluvoxamine. Furthermore, the effect of MDMA on L-DOPA-induced motor activity and dyskinesia was partially inhibited by 5-HT1a/b antagonists. The ability of MDMA to inhibit dyskinesia results from its broad spectrum of action on 5-HT systems. Serotoninergic receptors appear to play an important modulatory role in l-DOPA-induced dyskinesia, and this study may provide a framework for the use of serotoninergic agents in the treatment of L-DOPA-induced dyskinesia. PMID- 14534245 TI - Impaired long-term memory and NR2A-type NMDA receptor-dependent synaptic plasticity in mice lacking c-Fos in the CNS. AB - The immediate early gene c-fos is part of the activator protein-1 transcription factor and has been postulated to participate in the molecular mechanisms of learning and memory. To test this hypothesis in vivo, we generated mice with a nervous system-specific c-fos knock-out using the Cre-loxP system. Adult mice lacking c-Fos in the CNS (c-fosDeltaCNS) showed normal general and emotional behavior but were specifically impaired in hippocampus-dependent spatial and associative learning tasks. These learning deficits correlated with a reduction of long-term potentiation (LTP) in hippocampal CA3-CA1 synapses. The magnitude of LTP was restored by a repeated tetanization procedure, suggesting impaired LTP induction in c-fosDeltaCNS mice. This rescue was blocked by a selective inhibitor of NR2B-type NMDA receptors. This blockade was compensated in wild-type mice by NR2A-type NMDA receptor-activated signaling pathways, thus indicating that these pathways are compromised in c-fosDeltaCNS mice. In summary, our data suggest a role for c-Fos in hippocampus-dependent learning and memory as well as in NMDA receptor-dependent LTP formation. PMID- 14534246 TI - Quantitative estimate of synaptic inputs to striatal neurons during up and down states in vitro. AB - Up states are prolonged membrane potential depolarizations critical for synaptic integration and action potential generation in cortical and striatal neurons. They commonly result from numerous concurrent synaptic inputs, whereas neurons reside in a down state when synaptic inputs are few. By quantifying the composition, frequency, and amplitude of synaptic inputs for both states, we provide important constraints for state transitions in striatal network dynamics. Up and down states occur naturally in cortex-striatum-substantia nigra cocultures, which were used as an in vitro model in the present study. Spontaneous synaptic inputs during down states were extracted automatically in spiny projection neurons and fast spiking interneurons of the striatum using a newly developed computer algorithm. Consistent with a heterogeneous population of synaptic inputs, PSPs and PSCs showed no correlation in amplitude and rise time and occurred at relatively low frequencies of 10-40 Hz during the down state. The number of synaptic inputs during up states, estimated from the up-state charge and the unitary charge of down-state PSCs, was 217 +/- 44. Given the average up state duration of 284 +/- 34 msec, synaptic input frequency was approximately 800 Hz during up-states for both neuronal types. Many down-state events reversed at the chloride reversal potential and were blocked by GABA(A) antagonists. The high correlation between up- and down-state reversal potential suggests that despite these drastic changes in synaptic input frequency, the ratio of inhibitory to excitatory currents is similar during both states. PMID- 14534247 TI - sup-9, sup-10, and unc-93 may encode components of a two-pore K+ channel that coordinates muscle contraction in Caenorhabditis elegans. AB - Genetic studies of sup-9, unc-93, and sup-10 strongly suggest that these genes encode components of a multi-subunit protein complex that coordinates muscle contraction in Caenorhabditis elegans. We cloned sup-9 and sup-10 and found that they encode a two-pore K+ channel and a novel transmembrane protein, respectively. We also found that UNC-93 and SUP-10 colocalize with SUP-9 within muscle cells, and that UNC-93 is a member of a novel multigene family that is conserved among C. elegans, Drosophila, and humans. Our results indicate that SUP 9 and perhaps other two-pore K+ channels function as multiprotein complexes, and that UNC-93 and SUP-10 likely define new classes of ion channel regulatory proteins. PMID- 14534248 TI - Encoding of whisker vibration by rat barrel cortex neurons: implications for texture discrimination. AB - Rats, using their whiskers, have excellent capabilities in texture discrimination. What is the representation of texture in rat somatosensory cortex? We hypothesize that as rats "whisk" over a surface, the spatial frequency of a grooved or pebbled texture is converted to a temporal frequency of whisker vibration. Surface features such as groove depth or grain size modulate the amplitude of this vibration. Validation of the hypothesis depends on showing that vibration parameters have distinct neuronal representations in cortex. To test this, we delivered sinusoidal vibrations to the whisker shaft and analyzed cortical neuronal activity. Seven amplitudes and seven frequencies were combined to construct 49 stimuli while recording activity through a 10 x 10 microelectrode array inserted into the middle layers of barrel cortex. We find that cortical neurons do not explicitly encode vibration frequency (f) or amplitude (A) by any coding measure (average spike counts over different time windows, spike timing patterns in the peristimulus time histograms or in autocorrelograms). Instead, neurons explicitly encode the product of frequency and amplitude, which is proportional to the mean speed of the vibration. The quantity Af is an invariant because neuronal response encodes this feature independently of the values of the individual terms A and f. This was true across a wide time scale of firing rate measurements, from 5 to 500 msec. We conclude that vibration kinetics are rapidly and reliably encoded in the firing rate of cortical ensembles. Therefore, the cortical representation of vibration speed could underlie texture discrimination. PMID- 14534249 TI - Illusory sound perception in macaque monkeys. AB - In most natural listening environments, noise occludes objects of interest, and it would be beneficial for an organism to correctly identify those objects. When a sound of interest ("foreground" sound) is interrupted by a loud noise, subjects perceive the entire sound, even if the noise was intense enough to completely mask a part of it. This phenomenon can be exploited to create an illusion: when a silent gap is introduced into the foreground and high-intensity noise is superimposed into the gap, subjects report the foreground as continuing through the noise although that portion of the foreground was deleted. This phenomenon, referred to as auditory induction or amodal completion, is conceptually similar to visual induction, fill-in, illusory motion, and illusory contours. Two rhesus macaque monkeys performed a task designed to assess auditory induction. They were trained to discriminate complete stimuli from those containing a silent gap in the presence of two types of noise. Interrupting noise temporally coincided only with the gap, and in humans this causes induction. Surrounding noise temporally encompassed the entire foreground, and in humans this causes masking without auditory induction. Consistent with previous human psychophysical results, macaques showed better performance with surrounding masking noise than interrupting noise designed to elicit induction. These and other control experiments provide evidence that primates may share a general mechanism to perceptually complete missing sounds. PMID- 14534250 TI - Neuroprotective activities of sodium valproate in a murine model of human immunodeficiency virus-1 encephalitis. AB - Human immunodeficiency virus-1 (HIV-1) infection of the nervous system can result in neuroinflammatory events leading first to neuronal dysfunction then to cognitive and behavioral impairments in infected people. The multifaceted nature of the disease process, commonly called HIV-1-associated dementia (HAD), provides a number of adjunctive therapeutic opportunities. One proposed adjunctive therapy is sodium valproate (VPA), an anticonvulsant known to promote neurite outgrowth and increase beta-catenin through inhibiting glycogen synthase kinase 3beta activity and tau phosphorylation. We now show that VPA treatment of rat cortical neurons exposed to HIV-1 gp120 prevents resultant neurotoxic activities. This includes the induction of significant neurite outgrowth and microtubule associated protein 2 (MAP-2) and neuron-specific nuclear protein (NeuN) antigens in affected neuronal cell bodies and processes. Similarly, VPA protects severe combined immunodeficient (SCID) mice against the neurodegeneration of HIV-1ADA infected monocyte-derived macrophages (MDMs). In SCID mice with HIV-1 MDM-induced encephalitis, VPA treatment significantly reduced neuronal phosphorylatedbeta catenin and tau without affecting HIV-1 replication or glial activation. We conclude that VPA protects neurons against HIV-1 infected MDM neurotoxicity, possibly through its effects on the phosphorylation of tau and beta-catenin. The use of VPA as an adjuvant in treatment of human HAD is being pursued. PMID- 14534252 TI - Brain activation during human male ejaculation. AB - Brain mechanisms that control human sexual behavior in general, and ejaculation in particular, are poorly understood. We used positron emission tomography to measure increases in regional cerebral blood flow (rCBF) during ejaculation compared with sexual stimulation in heterosexual male volunteers. Manual penile stimulation was performed by the volunteer's female partner. Primary activation was found in the mesodiencephalic transition zone, including the ventral tegmental area, which is involved in a wide variety of rewarding behaviors. Parallels are drawn between ejaculation and heroin rush. Other activated mesodiencephalic structures are the midbrain lateral central tegmental field, zona incerta, subparafascicular nucleus, and the ventroposterior, midline, and intralaminar thalamic nuclei. Increased activation was also present in the lateral putamen and adjoining parts of the claustrum. Neocortical activity was only found in Brodmann areas 7/40, 18, 21, 23, and 47, exclusively on the right side. On the basis of studies in rodents, the medial preoptic area, bed nucleus of the stria terminalis, and amygdala are thought to be involved in ejaculation, but increased rCBF was not found in any of these regions. Conversely, in the amygdala and adjacent entorhinal cortex, a decrease in activation was observed. Remarkably strong rCBF increases were observed in the cerebellum. These findings corroborate the recent notion that the cerebellum plays an important role in emotional processing. The present study for the first time provides insight into which regions in the human brain play a primary role in ejaculation, and the results might have important implications for our understanding of how human ejaculation is brought about, and for our ability to improve sexual function and satisfaction in men. PMID- 14534253 TI - Cochlear phase and amplitude retrieved from the auditory nerve at arbitrary frequencies. AB - It is currently impossible to mechanically measure the overall vibration pattern of the intact mammalian cochlea because of its inaccessibility and vulnerability. At first sight, data from the auditory nerve are a poor substitute because of their limited temporal resolution. The nonlinear character of neural coding, however, causes low-frequency interactions among the components of multitone stimuli. We designed a novel stimulus for which these interactions take a particularly systematic form, and we recorded the response of the auditory nerve to this stimulus. A careful analysis of interactions in the data allowed us to reconstruct frequency transfer functions (both their amplitude and their phase) at multiple points spanning the entire length of the cochlea. The generic character of our stimuli and analysis suggests its wider use in nonlinear system analysis, particularly in those instances in which limitations in temporal resolution restrict the use of customary methods. PMID- 14534251 TI - Dorsal horn neurons firing at high frequency, but not primary afferents, release opioid peptides that produce micro-opioid receptor internalization in the rat spinal cord. AB - To determine what neural pathways trigger opioid release in the dorsal horn, we stimulated the dorsal root, the dorsal horn, or the dorsolateral funiculus (DLF) in spinal cord slices while superfusing them with peptidase inhibitors to prevent opioid degradation. Internalization of mu-opioid receptors (MOR) and neurokinin 1 receptors (NK1R) was measured to assess opioid and neurokinin release, respectively. Dorsal root stimulation at low, high, or mixed frequencies produced abundant NK1R internalization but no MOR internalization, indicating that primary afferents do not release opioids. Moreover, capsaicin and NMDA also failed to produce MOR internalization. In contrast, dorsal horn stimulation elicited MOR internalization that increased with the frequency, being negligible at <10 Hz and maximal at 500 Hz. The internalization was abolished by the MOR antagonist D-Phe Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2 (CTAP), in the presence of low Ca2+ and by the Na+ channel blocker lidocaine, confirming that it was caused by opioid release and neuronal firing. DLF stimulation in "oblique" slices (encompassing the DLF and the dorsal horn of T11-L4) produced MOR internalization, but only in areas near the stimulation site. Moreover, cutting oblique slices across the dorsal horn (but not across the DLF) eliminated MOR internalization in areas distal to the cut, indicating that it was produced by signals traveling in the dorsal horn and not via the DLF. These findings demonstrate that some dorsal horn neurons release opioids when they fire at high frequencies, perhaps by integrating signals from the rostral ventromedial medulla, primary afferents, and other areas of the spinal cord. PMID- 14534254 TI - Decreased temporal precision of auditory signaling in Kcna1-null mice: an electrophysiological study in vivo. AB - The voltage-gated potassium (Kv) channel subunit Kv1.1, encoded by the Kcna1 gene, is expressed strongly in the ventral cochlear nucleus (VCN) and the medial nucleus of the trapezoid body (MNTB) of the auditory pathway. To examine the contribution of the Kv1.1 subunit to the processing of auditory information, in vivo single-unit recordings were made from VCN neurons (bushy cells), axonal endings of bushy cells at MNTB cells (calyces of Held), and MNTB neurons of Kcna1 null (-/-) mice and littermate control (+/+) mice. Thresholds and spontaneous firing rates of VCN and MNTB neurons were not different between genotypes. At higher sound intensities, however, evoked firing rates of VCN and MNTB neurons were significantly lower in -/- mice than +/+ mice. The SD of the first-spike latency (jitter) was increased in VCN neurons, calyces, and MNTB neurons of -/- mice compared with +/+ controls. Comparison along the ascending pathway suggests that the increased jitter found in -/- MNTB responses arises mostly in the axons of VCN bushy cells and/or their calyceal terminals rather than in the MNTB neurons themselves. At high rates of sinusoidal amplitude modulations, -/- MNTB neurons maintained high vector strength values but discharged on significantly fewer cycles of the amplitude-modulated stimulus than +/+ MNTB neurons. These results indicate that in Kcna1-null mice the absence of the Kv1.1 subunit results in a loss of temporal fidelity (increased jitter) and the failure to follow high frequency amplitude-modulated sound stimulation in vivo. PMID- 14534255 TI - A receptor-like inositol lipid phosphatase is required for the maturation of developing cochlear hair bundles. AB - A screen for protein tyrosine phosphatases (PTPs) expressed in the chick inner ear yielded a high proportion of clones encoding an avian ortholog of protein tyrosine phosphatase receptor Q (Ptprq), a receptor-like PTP. Ptprq was first identified as a transcript upregulated in rat kidney in response to glomerular nephritis and has recently been shown to be active against inositol phospholipids. An antibody to the intracellular domain of Ptprq, anti-Ptprq, stains hair bundles in mice and chicks. In the chick ear, the distribution of Ptprq is almost identical to that of the 275 kDa hair-cell antigen (HCA), a component of hair-bundle shaft connectors recognized by a monoclonal antibody (mAb) that stains inner-ear hair bundles and kidney glomeruli. Furthermore, anti Ptprq immunoblots a 275 kDa polypeptide immunoprecipitated by the anti-HCA mAb from the avian inner ear, indicating that the HCA and Ptprq are likely to be the same molecule. In two transgenic mouse strains with different mutations in Ptprq, anti-Ptprq immunoreactivity cannot be detected in the ear. Shaft connectors are absent from mutant vestibular hair bundles, but the stereocilia forming the hair bundle are not splayed, indicating that shaft connectors are not necessary to hold the stereocilia together; however, the mice show rapid postnatal deterioration in cochlear hair-bundle structure, associated with smaller than normal transducer currents with otherwise normal adaptation properties, a progressive loss of basal-coil cochlear hair cells, and deafness. These results reveal that Ptprq is required for formation of the shaft connectors of the hair bundle, the normal maturation of cochlear hair bundles, and the long-term survival of high-frequency auditory hair cells. PMID- 14534256 TI - Glutamate receptor subunit 2 Serine 880 phosphorylation modulates synaptic transmission and mediates plasticity in CA1 pyramidal cells. AB - The cytoplasmic C termini of AMPA receptor subunits contain PDZ (postsynaptic density 95/Discs large/zona occludens 1) ligand domains that can control their synaptic trafficking during plasticity. The glutamate receptor subunit 2 (GluR2) PDZ ligand domain can be phosphorylated at serine 880 (S880), and this disrupts interactions with GRIP/ABP (glutamate receptor-interacting protein/AMPA-binding protein) but not with PICK1 (PKC-interacting protein 1). Here, the impact of GluR2 S880 phosphorylation on synaptic transmission and plasticity was explored by expressing, in hippocampal slice cultures, GluR2 subunits containing point mutations that mimic or prevent phosphorylation at this residue. Our results indicate that mimicking GluR2 S880 phosphorylation excludes these receptors from synapses, depresses transmission, and partially occludes long-term depression (LTD). Conversely, mutations that prevent phosphorylation reduce LTD. Disruption of the interaction between GluR2 and GRIP/ABP by S880 phosphorylation may thus facilitate removal of synaptic AMPA receptors and mediate some forms of activity dependent synaptic depression. PMID- 14534257 TI - The transmembrane semaphorin Sema4D/CD100, an inhibitor of axonal growth, is expressed on oligodendrocytes and upregulated after CNS lesion. AB - Semaphorins are a family of secreted and membrane-bound proteins, known to regulate axonal pathfinding. Sema4D, also called CD100, was first isolated in the immune system where it is involved in B and T cell activation. We found that in the mouse, Sema4D is expressed in cells throughout the CNS white matter, with a peak during the myelination period. Double-labeling experiments with different markers of oligodendrocyte lineage such as olig1, olig2, platelet-derived growth factor receptor alpha, and proteolipid protein showed that Sema4D was expressed selectively by oligodendrocytes and myelin. The presence of Sema4D in myelin was confirmed using Western blot. Sema4D expression in myelinating oligodendrocytes was further observed using neuron-oligodendrocyte cocultures. Moreover, using stripe assay, we found that Sema4D is strongly inhibitory for postnatal sensory and cerebellar granule cell axons. This prompted us to examine whether Sema4D expression is modified after CNS injury. At 8 d after spinal cord lesions, Sema4D expression was strongly upregulated in oligodendrocytes at the periphery of the lesion. Sema4D-positive cells were not colabeled with the astrocyte marker GFAP, with the microglial and macrophagic marker isolectin B4, or with NG2, a marker of oligodendrocyte precursors. This upregulation was transient because from 1 month after the lesion, Sema4D expression was back to its normal level. These results indicate that Sema4D is a novel inhibitory factor for axonal regeneration expressed in myelin. PMID- 14534258 TI - Brain structures differ between musicians and non-musicians. AB - From an early age, musicians learn complex motor and auditory skills (e.g., the translation of visually perceived musical symbols into motor commands with simultaneous auditory monitoring of output), which they practice extensively from childhood throughout their entire careers. Using a voxel-by-voxel morphometric technique, we found gray matter volume differences in motor, auditory, and visual spatial brain regions when comparing professional musicians (keyboard players) with a matched group of amateur musicians and non-musicians. Although some of these multiregional differences could be attributable to innate predisposition, we believe they may represent structural adaptations in response to long-term skill acquisition and the repetitive rehearsal of those skills. This hypothesis is supported by the strong association we found between structural differences, musician status, and practice intensity, as well as the wealth of supporting animal data showing structural changes in response to long-term motor training. However, only future experiments can determine the relative contribution of predisposition and practice. PMID- 14534259 TI - Fast synaptic currents in Drosophila mushroom body Kenyon cells are mediated by alpha-bungarotoxin-sensitive nicotinic acetylcholine receptors and picrotoxin sensitive GABA receptors. AB - The mushroom bodies, bilaterally symmetric regions in the insect brain, play a critical role in olfactory associative learning. Genetic studies in Drosophila suggest that plasticity underlying acquisition and storage of memory occurs at synapses on the dendrites of mushroom body Kenyon cells (Dubnau et al., 2001). Additional exploration of the mechanisms governing synaptic plasticity contributing to these aspects of olfactory associative learning requires identification of the receptors that mediate fast synaptic transmission in Kenyon cells. To this end, we developed a culture system that supports the formation of excitatory and inhibitory synaptic connections between neurons harvested from the central brain region of late-stage Drosophila pupae. Mushroom body Kenyon cells are identified as small-diameter, green fluorescent protein-positive (GFP+) neurons in cultures from OK107-GAL4;UAS-GFP pupae. In GFP+ Kenyon cells, fast EPSCs are mediated by alpha-bungarotoxin-sensitive nicotinic acetylcholine receptors (nAChRs). The miniature EPSCs have rapid rise and decay kinetics and a broad, positively skewed amplitude distribution. Fast IPSCs are mediated by picrotoxin-sensitive chloride conducting GABA receptors. The miniature IPSCs also have a rapid rate of rise and decay and a broad amplitude distribution. The vast majority of spontaneous synaptic currents in the cultured Kenyon cells are mediated byalpha-bungarotoxin-sensitive nAChRs or picrotoxin-sensitive GABA receptors. Therefore, these receptors are also likely to mediate synaptic transmission in Kenyon cells in vivo and to contribute to plasticity during olfactory associative learning. PMID- 14534260 TI - Signaling at the gliovascular interface. AB - Advances in fluorescent calcium indicating dyes over the past decade have identified calcium signaling as the tool by which astrocytes communicate among themselves and with neighboring neurons. Studies of astrocyte-neuron interactions have shown that calcium signaling is a potent modulator of the strength of both excitatory and inhibitory synapses. The concept that astrocytes possess a mechanism for rapid cell communication has not been incorporated, however, into the supportive functions of astrocytes. Because many of the classical tasks of astrocytes are linked to the blood-brain barrier, we have here examined the expression of proteins required for calcium signaling in their vascular end-foot processes. The gap junction protein, Cx43, was expressed intensively around the vessels interconnecting astrocytic end-foot processes. These gap junctions permitted diffusion of Lucifer yellow, specifically along the path of glial end feet apposed to the vessel wall. The purinergic receptors, P2Y(2) and P2Y(4), were also strongly expressed at the gliovascular interface and colocalized with GFAP around larger vessels in cortex. Multiphoton imaging of freshly prepared brain slices loaded with Fluo-4/AM revealed that ATP mobilized cytosolic calcium in astrocytic end feet, whereas electrical stimulation triggered calcium waves propagating along the vessel wall. Brain endothelial cells and pericytes were physically separated from astrocytes by the basal lamina and responded only weakly to ATP. These observations identify astrocytic end-foot processes plastered at the vessel wall as a center for purinergic signaling. It is speculated that calcium signaling may play a role in astrocytic functions related to the blood-brain barrier, including blood flow regulation, metabolic trafficking, and water homeostasis. PMID- 14534262 TI - Connecting the dots between animal and human studies of locomotion. Focus on "Infants adapt their stepping to repeated trip-inducing stimuli". PMID- 14534263 TI - Cortex controls multisensory depression in superior colliculus. AB - Multisensory depression is a fundamental index of multisensory integration in superior colliculus (SC) neurons. It is initiated when one sensory stimulus (auditory) located outside its modality-specific receptive field degrades or eliminates the neuron's responses to another sensory stimulus (visual) presented within its modality-specific receptive field. The present experiments demonstrate that the capacity of SC neurons to engage in multisensory depression is strongly dependent on influences from two cortical areas (the anterior ectosylvian and rostral lateral suprasylvian sulci). When these cortices are deactivated, the ability of SC neurons to synthesize visual-auditory inputs in this way is compromised; multisensory responses are disinhibited, becoming more vigorous and in some cases indistinguishable from responses to the visual stimulus alone. Although obtaining a more robust multisensory SC response when cortex is nonfunctional than when it is functional may seem paradoxical, these data may help explain previous observations that the loss of these cortical influences permits visual orientation behavior in the presence of a normally disruptive auditory stimulus. PMID- 14534264 TI - Differential thermosensitivity of sensory neurons in the guinea pig trigeminal ganglion. AB - Intracellular recordings were employed to study the effects of temperature on membrane properties and excitability in sensory neurons of the intact guinea pig trigeminal ganglion (TG) maintained in vitro. Neurons were classified according to the shape and duration of the action potential into F (short-duration, fast spike) and S (long duration, slow spike with a "hump") types. Most type F (33/34) neurons had axons with conduction velocities >1.5 m/s, while only 30% (6/23) of type S neurons reached these conduction speeds suggesting differences in myelination. Cooling reduced axonal conduction velocity and prolonged spike duration in both neuronal types. In F-type neurons with strong inward rectification. cooling also increased the excitability, augmenting the input resistance and reducing the current firing threshold. These effects were not observed in S-type neurons lacking inward rectification. In striking contrast to results obtained in cultured TG neurons, cooling or menthol did not induce firing in recordings from the acutely isolated ganglion. However, after application of submillimolar concentrations (100 microM) of the potassium channel blocker 4 aminopyridine (4-AP), 29% previously unresponsive neurons developed cold sensitivity. An additional 31% developed ongoing activity that was sensitive to temperature. Only neurons with strong inward rectification (mostly F-type) became thermosensitive. Cooling- and 4-AP-evoked firing were insensitive to intracellular application of 4-AP or somatic membrane hyperpolarization, suggesting that their action was most prominent at the level of the axon. The lack of excitatory actions of low temperature in the excised intact ganglion contrasts with the impulse discharges induced by cooling in trigeminal nerve terminals of the same species, suggesting a critical difference between cold transduction mechanisms at the level of the nerve terminals and the soma. PMID- 14534265 TI - Low-calcium epileptiform activity in the hippocampus in vivo. AB - It has been clearly established that nonsynaptic interactions are sufficient for generating epileptiform activity in brain slices. However, it is not known whether this type of epilepsy model can be generated in vivo. In this paper we investigate low-calcium nonsynaptic epileptiform activity in an intact hippocampus. The calcium chelator EGTA was used to lower [Ca2+]o in the hippocampus of urethane anesthetized rats. Spontaneous and evoked field potentials in CA1 pyramidal stratum and in CA1 stratum radiatum were recorded using four-channel silicon recording probes. Three different types of epileptic activity were observed while synaptic transmission was gradually blocked by a decline in hippocampal [Ca2+]o. A short latency burst, named early-burst, occurred during the early period of EGTA application. Periodic slow-waves and a long latency high-frequency burst, named late-burst, were seen after synaptic transmission was mostly blocked. Therefore these activities appear to be associated with nonsynaptic mechanisms. Moreover, the slow-waves were similar in appearance to the depolarization potential shifts in vitro with low calcium. In addition, excitatory postsynaptic amino acid antagonists could not eliminate the development of slow-waves and late-bursts. The slow-waves and late-bursts were morphologically similar to electrographic seizure activity seen in patients with temporal lobe epilepsy. These results clearly show that epileptic activity can be generated in vivo in the absence of synaptic transmission. This type of low calcium nonsynaptic epilepsy model in an intact hippocampus could play an important role in revealing additional mechanisms of epilepsy disorders and in developing novel anti-convulsant drugs. PMID- 14534266 TI - Postnatal development of cochlear function in the mustached bat, Pteronotus parnellii. AB - Postnatal development of the mustached bat's cochlea was studied by measuring cochlear microphonic and compound action potentials. In adults, a cochlear resonance is involved in enhanced tuning to the second harmonic constant frequency component (CF2) of their echolocation calls at approximately 61 kHz This resonance is present immediately after birth in bats that do not yet echolocate. Its frequency is lower (46 kHz) and the corresponding threshold minimum of cochlear microphonic potentials is broader than in adults. Long lasting ringing of the cochlear microphonic potential after tone stimulus offset that characterizes the adult auditory response close to CF2 is absent in newborns. In the course of the first 5 postnatal weeks, there is a concomitant upward shift of CF2 and the frequency of cochlear threshold minima. Up to the end of the third postnatal week, sensitivity of auditory threshold minima and the Q value of the cochlear resonance increase at a fast rate. Between 2 and 4 wk of age, two cochlear microphonic threshold minima are found consistently in the CF2 range that differ in their level-dependent dynamic growth behavior and are 1.5 5.7 kHz apart from each other. In older animals, there is a single minimum that approaches adult tuning in its sharpness. The data provide evidence to show that during maturation of the cochlea, the frequency and the sensitivity of the threshold minimum associated with CF2 increases and that these increases are associated with the fusion of two resonances that are partly dissociated in developing animals. PMID- 14534267 TI - Development of echolocation calls in the mustached bat, Pteronotus parnellii. AB - Adult mustached bats employ Doppler-sensitive sonar to hunt fluttering prey insects in acoustically cluttered habitats. The echolocation call consists of 4-5 harmonics, each composed of a long constant frequency (CF) component flanked by brief frequency modulations (FM). The 2nd harmonic CF component (CF2) at 61 kHz is the most intense, and analyzed by an exceptionally sharply tuned auditory system. The maturation of echolocation calls and the development of Doppler-shift compensation was studied in Cuba where large maternity colonies are found in hot caves. In the 1st postnatal week, infant bats did not echolocate spontaneously but could be induced to vocalize CF-FM signals by passive body motion. The CF2 frequency emitted by the smallest specimens was at 48 kHz (i.e., 0.4 octaves lower than the adult signal). CF-FM signals were spontaneously produced in the 2nd postnatal week at a CF2 frequency of 52 kHz. The CF2 frequencies of induced and spontaneous calls shifted upward to reach a value of 60.5 kHz in the 5th postnatal week. Standard deviations of CF2 frequency were large (up to +/-1.5 kHz) in the youngest bats and dropped to values of +/-250 Hz at the end of the 3rd postnatal week. Some individuals in the 4th and 5th postnatal weeks emitted with adultlike frequency precision of about +/-100 Hz. In the youngest bats, the 1st harmonic CF component (CF1) was up to 22 dB stronger than CF2. Adultlike relative levels of CF1 (-28 dB relative to CF2) were reached in the 5th postnatal week. In spontaneously emitted CF-FM calls, the duration of the CF2 component gradually increased with age from 5 ms to maximum values of 18 ms. Durations of the CF2 component in induced calls averaged 7 +/- 2.6 ms in the 1st postnatal week and 8.2 +/- 1.5 ms in the 5th postnatal week. There were no age-related changes in duration of the terminal FM sweep (3 +/- 0.4 ms) in both induced and spontaneous calls. The magnitude of the terminal FM sweep in spontaneous calls was not correlated with age (mean 13.5 +/- 2 kHz). Values for induced calls slightly increased with age from 11 +/- 2 to 13 +/- 2 kHz. The emission rate of induced CF-FM signals increased with age from values of 2.5 +/- 2 to 17 +/- 5 pulses/s. Values for spontaneously emitted calls were 4.4 +/- 3 and 9 +/- 4.5 pulses/s, respectively. Doppler-shift compensation, as tested in the pendulum task, emerged during the 4th postnatal week in young bats that were capable of very brief active flights, but before the time of active foraging outside the cave. PMID- 14534268 TI - Suppression of corticospinal excitability during negative motor imagery. AB - To investigate the effect of negative motor imagery on corticospinal excitability, we performed transcranial magnetic stimulation (TMS) studies in seven healthy subjects during imagination of suppressing movements. Subjects were asked to imagine suppression of TMS-induced twitching movement of their nondominant left hands by attempting to increase the amount of relaxation after receiving an auditory NoGo cue (negative motor imagery), but to imagine squeezing hands after a Go cue (positive motor imagery). Single- and paired-pulse TMS were triggered at 2 s after Go or NoGo cues. Motor-evoked potentials (MEPs) were recorded in the first dorsal interosseus (FDI), abductor pollicis brevis (APB), and abductor digiti minimi (ADM) muscles of the left hand. Paired-pulse TMS with subthreshold conditioning stimuli at interstimulus intervals of 2 (short intracortical inhibition) and 15 ms (intracortical facilitation) and that with suprathreshold conditioning stimuli at interstimulus interval of 80 ms (long intracortical inhibition) were performed in both negative motor imagery and control conditions. Compared with the control state (no imagination), MEP amplitudes of FDI (but not APB and ADM) were significantly suppressed in negative motor imagery, but those from all three muscles were unchanged during positive motor imagery. F-wave responses (amplitudes and persistence) were unchanged during both negative and positive motor imagery. During negative motor imagery, resting motor threshold was significantly increased, but short and long intracortical inhibition and intracortical facilitation were unchanged. The present results demonstrate that excitatory corticospinal drive is suppressed during imagination of suppressing movements. PMID- 14534269 TI - The role of the corpus callosum in the coupling of bimanual isometric force pulses. AB - Two split-brain patients, a patient with callosal agenesis, and 6 age-matched control participants were tested on a bimanual force production task. The participants produced isometric responses with their index fingers, attempting to match the target force specified by a visual stimulus. On unimanual trials, the stimuli were presented in either the left or right visual field and the response was made with the ipsilateral hand. On bimanual trials, two stimuli were presented, one on each side, and the target forces could be either identical or different. Bimanual responses of the control subjects showed strong evidence of coupling. Forces produced by one hand were influenced by the forces produced by the other hand with positive correlations observed for all target force combinations. These assimilation effects and correlations were greatly attenuated in the acallosal group, with similar results observed for the split-brain patients and participant with callosal agenesis. Furthermore, the processes involved in selecting and planning the two responses occurred independently in the acallosal group; in contrast to the controls, the three acallosal participants exhibited no differences in reaction times or accuracy between bimanual trials in which the two target forces were the same or different. We also found a striking temporal desynchronization of the responses in the split brain patients, indicating that in this context, temporal coupling is impaired after callosotomy. These results are congruent with the hypothesis that interference related to response selection and planning of bimanual force pulses arises from callosal interactions. PMID- 14534270 TI - Retinogeniculate synaptic properties controlling spike number and timing in relay neurons. AB - Retinal ganglion cells (RGC) transmit visual signals to thalamocortical relay neurons in the lateral geniculate nucleus via retinogeniculate synapses. Relay neuron spike patterns do not simply reflect those of RGCs, but the mechanisms controlling this transformation are not well understood. We therefore examined synaptic properties controlling the strength and precision of relay neuron firing in mouse (p28-33) brain slices using physiological stimulation patterns and a combination of current clamp and dynamic clamp. In tonic mode (-55 mV), activation of single RGC inputs elicited stereotyped responses in a given neuron. In contrast, responses in different neurons varied from unreliable, to faithfully following, to a gain in the number of spikes. Dynamic clamp experiments indicated these different responses primarily reflected variability in the amplitudes of the N-methyl-d-aspartate (NMDA) and AMPA components. Each of these components played a distinct role in transmission. The AMPA component evoked a single precisely timed, short-latency spike per stimulus, but efficacy decreased during repetitive stimulation due to desensitization and depression. The NMDA component elicited longer-latency spikes and multiple spikes per stimulus and became more effective during repetitive stimuli that led to NMDA current summation. We found that in burst mode (-75 mV), where low-threshold calcium spikes are activated, AMPA and NMDA components and synaptic plasticity influenced spike number, but no combination enabled relay cells to faithfully follow the stimulus. Thus the characteristics of AMPA and NMDA currents, the ratio of these currents and use dependent plasticity interact to shape how RGC activity is conveyed to relay neurons. PMID- 14534271 TI - Changes in segmental and motor cortical output with contralateral muscle contractions and altered sensory inputs in humans. AB - Motor or sensory activity in one arm can affect the other arm. We tested the hypothesis that a voluntary contraction can affect the motor pathway to the contralateral homologous muscle and investigated whether alterations in sensory input might mediate such effects. Responses to transcranial magnetic stimulation [motor-evoked potentials (MEPs)], stimulation of the descending tracts [cervicomedullary MEPs (CMEPs)], and peripheral nerve stimulation (H-reflex) were recorded from the relaxed right flexor carpi radialis (FCR), while the left arm underwent unilateral interventions (5 s duration) that included voluntary contraction, muscle contraction evoked through percutaneous stimulation, tendon vibration, and cutaneous and mixed nerve stimulation. During moderate to strong voluntary wrist flexion on the left, MEPs in the right FCR increased, CMEPs were unaffected, and the H-reflex was depressed. These results are consistent with an increase in excitability of the motor cortex, no effect on the motoneuron pool, and presynaptic inhibition of Ia afferents. In contrast, percutaneous muscle stimulation facilitated both MEPs and the H-reflex. However, muscle contraction produced by a combination of voluntary effort and electrical stimulation also reduced the contralateral H-reflex. After voluntary contractions, the H-reflex remained depressed for 35 s, but after stimulation-evoked contractions, it rapidly returned to baseline. Under both conditions, MEPs recovered rapidly. After voluntary contractions, CMEPs were also depressed for approximately 10 s despite their lack of change during contractions. Wrist tendon vibration (100 Hz) did not affect, and 20-Hz median nerve stimulation or forearm medial cutaneous nerve stimulation mildly facilitated, the H-reflex without affecting MEPs. Voluntary wrist extension, similarly to wrist flexion, increased MEPs and depressed H-reflexes. However, ankle dorsiflexion facilitated the H-reflex akin to the Jendrassik maneuver. These data suggest that a unilateral voluntary muscle contraction has contralateral effects at both cortical and segmental levels and that the segmental effects are not replicated by stimulated muscle contraction or by input from muscle spindles or non-nociceptive cutaneous afferents. PMID- 14534272 TI - NO enhances presynaptic currents during cerebellar mossy fiber-granule cell LTP. AB - Nitric oxide (NO) is a candidate retrograde messenger in long-term potentiation (LTP). The NO metabolic pathway is expressed in the cerebellar granule cell layer but its physiological role remained unknown. In this paper we have investigated the role of NO in cerebellar mossy fiber-granule cell LTP, which has postsynaptic N-methyl-d-aspartate (NMDA) receptor-dependent induction. Pre- and postsynaptic current changes were simultaneously measured by using extracellular focal recordings, and NO release was monitored with an electrochemical probe in P21 rat cerebellar slices. High-frequency mossy fiber stimulation induced LTP and caused a significant NO release (6.2 +/- 2.8 nM; n = 5) in the granular layer that was dependent on NMDA receptor as well as on nitric oxide synthase (NOS) activation. Preventing NO production by perfusing the NOS inhibitor 100 microM NG-nitro-l arginine (L-NNA), blocking extracellular NO diffusion by 10 microM MbO2, or inhibiting the NO target guanylyl cyclase (sGC) with 10 microM 1H [1,2,4]oxadiazolo[4,3-a]quinoxalin-1-dione (ODQ) prevented LTP. Moreover, the NO donor 10 microM 2-(N,N-diethylamino)-diazenolate-2-oxide.Na (DEA-NO) induced LTP, which was mutually occlusive with LTP generated by high-frequency stimulation, prevented by ODQ, and insensitive to NMDA channel blockade (50 microM APV + 25 microM 7-Cl-kyn) or interruption of mossy fiber stimulation. Thus NO is critical for LTP induction at the cerebellar mossy fiber-granule cell relay. Interestingly, LTP manipulations were accompanied by consensual changes in the presynaptic current, suggesting that NO acts as a retrograde signal-enhancing presynaptic terminal excitability. PMID- 14534273 TI - Temporal pattern recognition based on instantaneous spike rate coding in a simple auditory system. AB - Auditory pattern recognition by the CNS is a fundamental process in acoustic communication. Because crickets communicate with stereotyped patterns of constant frequency syllables, they are established models to investigate the neuronal mechanisms of auditory pattern recognition. Here we provide evidence that for the neural processing of amplitude-modulated sounds, the instantaneous spike rate rather than the time-averaged neural activity is the appropriate coding principle by comparing both coding parameters in a thoracic interneuron (Omega neuron ON1) of the cricket (Gryllus bimaculatus) auditory system. When stimulated with different temporal sound patterns, the analysis of the instantaneous spike rate demonstrates that the neuron acts as a low-pass filter for syllable patterns. The instantaneous spike rate is low at high syllable rates, but prominent peaks in the instantaneous spike rate are generated as the syllable rate resembles that of the species-specific pattern. The occurrence and repetition rate of these peaks in the neuronal discharge are sufficient to explain temporal filtering in the cricket auditory pathway as they closely match the tuning of phonotactic behavior to different sound patterns. Thus temporal filtering or "pattern recognition" occurs at an early stage in the auditory pathway. PMID- 14534274 TI - Short-term desensitization of G-protein-activated, inwardly rectifying K+ (GIRK) currents in pyramidal neurons of rat neocortex. AB - Whole cell recordings from acutely isolated rat neocortical pyramidal cells were performed to study the kinetics and the mechanisms of short-term desensitization of G-protein-activated, inwardly rectifying K+ (GIRK) currents during prolonged application (5 min) of baclofen, adenosine, or serotonin. Most commonly, desensitization of GIRK currents was characterized by a biphasic time course with average time constants for fast and slow desensitization in the range of 8 and 120 s, respectively. The time constants were independent of the agonist used to evoke the current. The biphasic time course was preserved in perforated-patch recordings, indicating that neither component of desensitization is attributable to cell dialysis. Desensitization of GIRK currents displayed a strong heterologous component in that application of a second agonist substantially reduced the responsiveness to a test agonist. Fast desensitization, but not slow desensitization, was lost in cells loaded with GDP, suggesting that the hydrolysis cycle of G proteins might underlie the initial, rapid current decline. Hydrolysis of phosphatidylinositol biphosphate is an unlikely candidate underlying short-term desensitization, because both components of desensitization were preserved in the presence of the phospholipase C inhibitor U73122. We conclude that short-term desensitization does neither result from receptor downregulation nor from altered channel gating but might involve modifications of the G-protein-dependent pathway that serves to translate receptor activation into channel opening. PMID- 14534275 TI - Human ocular pursuit during the transient disappearance of a visual target. AB - During the course of pursuing a moving target there are often periods of transient disappearance as it moves behind objects and surfaces. In experimental settings, eye velocity decays rapidly on the extinction of a moving target. However, eye velocity does not decay to zero if there is an expectation the target will reappear further along its trajectory. Increasing eye velocity to coincide with target reappearance could minimize the developing velocity error, but it remains to be empirically verified whether this can be achieved. The aim of the present study was to examine the influence of stimulus predictability, target velocity, and interstimulus interval (ISI) on ocular pursuit during the transient disappearance of a visual target. We confirmed that subjects (n = 9) did not maintain eye velocity close to target velocity for the duration of the ISI. In general, after an initial reduction in eye velocity the majority of subjects (n = 7) exhibited a significant increase before target reappearance. The timing of the velocity increase was not influenced by target velocity, stimulus predictability, or ISI. Consequently, for the 900-ms ISI the increase occurred too early and the eye was decelerating at the moment of target reappearance. These results are consistent with a reduction in gain being applied to the visuomotor drive when the target disappeared, followed by a reactivation in expectation of target reappearance. We modeled this process such that gain was modulated within a reafferent feedback system, hence preserving its output in the absence of negative visual feedback and enabling an anticipatory increase in eye velocity before expected target reappearance. PMID- 14534276 TI - Electrophysiological evidence of monosynaptic excitatory transmission between granule cells after seizure-induced mossy fiber sprouting. AB - Mossy fiber sprouting is a form of synaptic reorganization in the dentate gyrus that occurs in human temporal lobe epilepsy and animal models of epilepsy. The axons of dentate gyrus granule cells, called mossy fibers, develop collaterals that grow into an abnormal location, the inner third of the dentate gyrus molecular layer. Electron microscopy has shown that sprouted fibers from synapses on both spines and dendritic shafts in the inner molecular layer, which are likely to represent the dendrites of granule cells and inhibitory neurons. One of the controversies about this phenomenon is whether mossy fiber sprouting contributes to seizures by forming novel recurrent excitatory circuits among granule cells. To date, there is a great deal of indirect evidence that suggests this is the case, but there are also counterarguments. The purpose of this study was to determine whether functional monosynaptic connections exist between granule cells after mossy fiber sprouting. Using simultaneous recordings from granule cells, we obtained direct evidence that granule cells in epileptic rats have monosynaptic excitatory connections with other granule cells. Such connections were not obtained when age-matched, saline control rats were examined. The results suggest that indeed mossy fiber sprouting provides a substrate for monosynaptic recurrent excitation among granule cells in the dentate gyrus. Interestingly, the characteristics of the excitatory connections that were found indicate that the pathway is only weakly excitatory. These characteristics may contribute to the empirical observation that the sprouted dentate gyrus does not normally generate epileptiform discharges. PMID- 14534277 TI - Glutamatergic propagation of GABAergic seizure-like afterdischarge in the hippocampus in vitro. AB - Previous investigations have suggested that GABA may act actively as an excitatory mediator in the generation of seizure-like (ictal) or interictal epileptiform activity in several experimental models of temporal lobe epilepsy. However, it remains to be known whether or not such GABAergic excitation may participate in seizure propagation into neighboring cortical regions. In our in vitro study using mature rat hippocampal slices, we examined the cellular mechanism underlying synchronous propagation of seizure-like afterdischarge in the CA1 region, which is driven by depolarizing GABAergic transmission, into the adjacent subiculum region. Tetanically induced seizure-like afterdischarge was always preceded by a GABAergic, slow posttetanic depolarization in the pyramidal cells of the original seizure-generating region. In contrast, the slow posttetanic depolarization was no longer observed in the subicular pyramidal cells when the afterdischarge was induced in the CA1 region. Surgical cutting of axonal pathways through the stratum oriens and the alveus between the CA1 and the subiculum region abolished the CA1-generated afterdischarge in the subicular pyramidal cells. Intracellular loading of fluoride ions, a GABAA receptor blocker, into single subicular pyramidal cells had no inhibitory effect on the CA1-generated afterdischarge in the pyramidal cells. Furthermore, the CA1 generated afterdischarge in the subicular pyramidal cells was largely depressed by local application of glutamate receptor antagonists to the subiculum region during afterdischarge generation. The present results indicate that the excitatory GABAergic generation of seizure-like activity seems to be restricted to epileptogenic foci of origin in the seizure-like epilepsy model in vitro. PMID- 14534278 TI - Synaptically activated Cl- accumulation responsible for depolarizing GABAergic responses in mature hippocampal neurons. AB - It is known that GABA, a major inhibitory transmitter in the CNS, acts as an excitatory (or depolarizing) transmitter transiently after intense GABAA receptor activation in adult brains. The depolarizing effect is considered to be dependent on two GABAA receptor-permeable anions, chloride (Cl-) and bicarbonate (HCO3-). However, little is known about their spatial and temporal profiles during the GABAergic depolarization in postsynaptic neurons. In the present study, we show that the amplitude of synaptically induced depolarizing response was correlated with intracellular Cl- accumulation in the soma of mature hippocampal CA1 pyramidal cells, by using whole cell patch-clamp recording and Cl- imaging technique with a Cl- indicator 6-methoxy-N-ethylquinolinium iodide (MEQ). The synaptically activated Cl- accumulation was mediated dominantly through GABAA receptors. Basket cells, a subclass of fast-spiking interneurons innervating the somatic portion of the pyramidal cells, actually fired at high frequency during the Cl- accumulation accompanying the depolarizing responses. These results suggest synaptically activated GABAA-mediated Cl- accumulation may play a critical role in generation of an excitatory GABAergic response in the mature pyramidal cells receiving intense synaptic inputs. This may be the first demonstration of microscopic visualization of intracellular Cl- accumulation during synaptic activation. PMID- 14534279 TI - Saccades exhibit abrupt transition between reactive and predictive; predictive saccade sequences have long-term correlations. AB - To compensate for neural delays, organisms require predictive motor control. We investigated the transition between reaction and prediction in saccades (rapid eye movements) to periodically paced targets. Tracking at low frequencies (0.2 0.3 Hz) is reactive (eyes lag target) and at high frequencies (0.9-1.0 Hz) is predictive (eyes anticipate target); there is an abrupt rather than smooth transition between the two modes (a "phase transition," as found in bistable physical systems). These behaviors represent stable modes of the oculomotor control system, with attendant rapid switching between the neural pathways underlying the different modes. Furthermore, predictive saccades exhibit long term correlations (slow decay of the autocorrelation function, manifest as a 1/f alpha spectrum). This indicates that predictive trials are not independent. The findings have implications for the understanding of predictive motor control: predictive performance during a given trial is influenced by a feedback process that takes into account the latency of previous trials. PMID- 14534280 TI - Contribution of head movement to gaze command coding in monkey frontal cortex and superior colliculus. AB - Most of what we know about the neural control of gaze comes from experiments in head-fixed animals, but several "head-free" studies have suggested that fixing the head dramatically alters the apparent gaze command. We directly investigated this issue by quantitatively comparing head-fixed and head-free gaze trajectories evoked by electrically stimulating 52 sites in the superior colliculus (SC) of two monkeys and 23 sites in the supplementary eye fields (SEF) of two other monkeys. We found that head movements made a significant contribution to gaze shifts evoked from both neural structures. In the majority of the stimulated sites, average gaze amplitude was significantly larger and individual gaze trajectories were significantly less convergent in space with the head free to move. Our results are consistent with the hypothesis that head-fixed stimulation only reveals the oculomotor component of the gaze shift, not the true, planned goal of the movement. One implication of this finding is that when comparing stimulation data against popular gaze control models, freeing the head shifts the apparent coding of gaze away from a "spatial code" toward a simpler visual model in the SC and toward an eye-centered or fixed-vector model representation in the SEF. PMID- 14534281 TI - Static ocular counterroll is implemented through the 3-D neural integrator. AB - Static head roll about the naso-occipital axis is known to produce an opposite ocular counterroll with a gain of approximately 10%, but the purpose and neural mechanism of this response remain obscure. In theory counterroll could be maintained either by direct tonic vestibular inputs to motoneurons, or by a neurally integrated pulse, as observed in the saccade generator and vestibulo ocular reflex. When simulated together with ocular drift related to torsional integrator failure, the direct tonic input model predicted that the pattern of drift would shift torsionally as in ordinary counterroll, but the integrated pulse model predicted that the equilibrium position of torsional drift would be unaffected by head roll. This was tested experimentally by measuring ocular counterroll in 2 monkeys after injection of muscimol into the mesencephalic interstitial nucleus of Cajal. Whereas 90 degrees head roll produced a mean ocular counterroll of 8.5 degrees (+/-0.7 degrees SE) in control experiments, the torsional equilibrium position observed during integrator failure failed to counterroll, showing a torsional shift of only 0.3 degrees (+/-0.6 degrees SE). This result contradicted the direct tonic input model, but was consistent with models that implement counterroll by a neurally integrated pulse. PMID- 14534282 TI - Is autocrine ATP release required for activation of volume-sensitive chloride channels? PMID- 14534283 TI - Spatial organization of frequency response areas and rate/level functions in the developing AI. AB - The current study was conducted to extend our understanding of changes in spatial organization and response properties of cortical neurons in the developing mammalian forebrain. Extracellular multiunit responses to tones were recorded from a dense array of penetrations covering entire isofrequency contours in the primary auditory cortex (AI) of pentobarbital anesthetized kittens. Ages ranged from postnatal day 14 (P14), shortly after acquisition of normal auditory response thresholds, through postnatal day 111 (P111), when the kittens were largely mature. Spatial organization of the AI was tonotopically ordered by P14. The tonotopic gradient decreased with chronological maturation. At P14 the gradient was about 3.5 kHz/mm. By P111 it had declined to about 2.5 kHz/mm, so that the cortical region encompassing a fixed 3- to 15-kHz frequency range enlarged along its posterior-anterior dimension. Response properties of developing AI neurons changed in both frequency selectivity and intensity selectivity. The mean frequency tuning bandwidth increased with age. Initially, tuning bandwidths were narrow throughout the entire AI. With progressive maturation, broader bandwidths were observed in areas dorsal and ventral to a central region in which neurons remained narrowly tuned. The resulting spatial organization of tuning bandwidth was similar to that reported in adult cats. The majority of recording sites manifested nonmonotonic rate/level functions at all ages. However, the proportion of sites with monotonic rate/level functions increased with age. No spatial organization of rate/level functions (monotonic and nonmonotonic) was observed through P111. The relatively late development of bandwidth tuning in the AI compared with the early presence of tonotopic organization suggests that different developmental processes are responsible for structuring these two dimensions of acoustic selectivity. PMID- 14534284 TI - Lurcher mice exhibit potentiation of GABA(A)-receptor-mediated conductance in cerebellar nuclei neurons in close temporal relationship to Purkinje cell death. AB - In heterozygous Lurcher mice (Lc/+), the Purkinje cells (PCs) degenerate almost totally during postnatal development. On the other hand, their projection target, the deep cerebellar nuclei (DCN), shows few signs of degeneration and seems to play an important role in maintaining a residual cerebellar function in Lc/+. We asked whether the DCN in Lc/+ develop cellular adaptations allowing them to cope with the loss of GABAergic PC input. Using whole-cell patch-clamp recordings, we measured inhibitory postsynaptic currents from DCN of Lc/+ and wild-type mice (WT). In experiments on phenotypically striking Lc/+ studied well after the onset of the PC degeneration, we found enlarged average synaptic conductances (g(syn)) compared with WT. We next investigated postnatal mice before and after the onset of PC death. In younger animals 1200 microm, suggesting a temporal "suppression annulus" surrounding each initiation site. Within the annulus, the likelihood for initiating the next sIS was reduced. Suppression annulus did not have a noticeable change in the presence of GABA(b) antagonist, suggesting it did not depend on the GABA(b) inhibition. We also applied bicuculline locally to a spot of 800 x 800 microm(2) for approximately 45 min. During this period approximately 1000 sISs occurred within the spot. Bicuculline or picrotoxin was then applied to the entire craniotomy window. The pretreatment created an obvious cluster of initiation sites. Around this cluster, the suppression annulus became obvious in individual animals. Our results suggest that, in disinhibited cortex, epileptiform events were initiated from small sites. The initiation sites may cluster in an area with increased local activity. Surrounding each initiation site there may be a temporal suppression annulus. PMID- 14534287 TI - Optic flow signals in extrastriate area MST: comparison of perceptual and neuronal sensitivity. AB - The medial superior temporal area of extrastriate cortex (MST) contains signals selective for nonuniform patterns of motion often termed "optic flow." The presence of such tuning, however, does not necessarily imply involvement in perception. To quantify the relationship between these selective neuronal signals and the perception of optic flow, we designed a discrimination task that allowed us to simultaneously record neuronal and behavioral sensitivities to near threshold optic flow stimuli tailored to MST cells' preferences. In this two alternative forced-choice task, we controlled the salience of globally opposite patterns (e.g., expansion and contraction) by varying the coherence of the motion. Using these stimuli, we could both relate the sensitivity of neuronal signals in MST to the animal's behavioral sensitivity and also measure trial-by trial correlation between neuronal signals and behavioral choices. Neurons in MST showed a wide range of sensitivities to these complex motion stimuli. Many neurons had sensitivities equal or superior to the monkey's threshold. On the other hand, trial-by-trial correlation between neuronal discharge and choice ("choice probability") was weak or nonexistent in our data. Together, these results lead us to conclude that MST contains sufficient information for threshold judgments of optic flow; however, the role of MST activity in optic flow discriminations may be less direct than in other visual motion tasks previously described by other laboratories. PMID- 14534286 TI - Factors underlying bursting behavior in a network of cultured hippocampal neurons exposed to zero magnesium. AB - Factors contributing to reduced magnesium-induced neuronal action potential bursting were investigated in primary hippocampal cell culture at high and low culture density. In nominally zero external magnesium medium, pyramidal neurons from high-density cultures produced recurrent spontaneous action potential bursts superimposed on prolonged depolarizations. These bursts were partially attenuated by the NMDA receptor antagonist d-APV. Pharmacological analysis of miniature excitatory postsynaptic currents (EPSCs) revealed 2 components: one sensitive to d-APV and another to the AMPA receptor antagonist DNQX. The components were kinetically distinct. Participation of NMDA receptors in reduced magnesium induced synaptic events was supported by the localization of the NR1 subunit of the NMDA receptor with the presynaptic vesicular protein synaptophysin. Presynaptically, zero magnesium induced a significant increase in EPSC frequency likely attributable to increased neuronal hyperexcitability induced by reduced membrane surface charge screening. Mean quantal content was significantly increased in zero magnesium. Cells from low-density cultures did not exhibit action potential bursting in zero magnesium but did show increased EPSC frequency. Low-density neurons had less synaptophysin immunofluorescence and fewer active synapses as determined by FM1-43 analysis. These results demonstrate that multiple factors are involved in network bursting. Increased probability of transmitter release presynaptically, enhanced NMDA receptor-mediated excitability postsynaptically, and extent of neuronal interconnectivity contribute to initiation and maintenance of elevated network excitability. PMID- 14534288 TI - Acetylcholine increases intracellular Ca2+ via nicotinic receptors in cultured PDF-containing clock neurons of Drosophila. AB - Light entrains the biological clock both in adult and larval Drosophila melanogaster. The Bolwig organ photoreceptors most likely constitute one substrate for this light entrainment in larvae. Acetylcholine (ACh) has been suggested as the neurotransmitter in these photoreceptors, but there is no evidence that ACh signaling is involved in photic input onto circadian pacemaker neurons. Here we demonstrate that the putative targets of the Bolwig photoreceptors, the PDF-containing clock neurons (LNs), in the larval brain express functional ACh receptors (AChRs). With the use of GAL4-UAS-driven expression of green fluorescent protein (GFP), we were able to identify LNs in dissociated cell culture. After loading with the Ca(2+)-sensitive dye fura-2, we monitored changes in intracellular Ca(2+) levels ([Ca(2+)](i)) in GFP-marked LNs while applying candidate neurotransmitters. ACh induced transient increases in [Ca(2+)](i) at physiological concentrations. These increases were dependent on extracellular Ca(2+) and Na(+) and were likely caused by activation of voltage dependent Ca(2+) channels. Application of nicotinic and muscarinic agonists and antagonists showed that the AChRs on cultured LNs have a nicotinic pharmacology. Antibodies to several subunits of nicotinic AChRs (nAChRs) labeled the putative contact site of the Bolwig organ axon terminals with the dendrites of LNs, as well as dissociated LNs in culture. Our findings support a role of ACh as input factor onto the LNs and suggest that Ca(2+) is used as a second messenger mediating cholinergic input within the LNs. Experiments using a more general GAL4 UAS-driven expression of GFP showed that functional expression of nAChRs is a widespread phenomenon in peptidergic neurons. PMID- 14534289 TI - Response growth with sound level in auditory-nerve fibers after noise-induced hearing loss. AB - People with sensorineural hearing loss are often constrained by a reduced acoustic dynamic range associated with loudness recruitment; however, the neural correlates of loudness and recruitment are still not well understood. The growth of auditory-nerve (AN) activity with sound level was compared in normal-hearing cats and in cats with a noise-induced hearing loss to test the hypothesis that AN fiber rate-level functions are steeper in impaired ears. Stimuli included best frequency and fixed-frequency tones, broadband noise, and a brief speech token. Three types of impaired responses were observed. 1) Fibers with rate-level functions that were similar across all stimuli typically had broad tuning, consistent with outer-hair-cell (OHC) damage. 2) Fibers with a wide dynamic range and shallow slope above threshold often retained sharp tuning, consistent with primarily inner-hair-cell (IHC) damage. 3) Fibers with very steep rate-level functions for all stimuli had thresholds above approximately 80 dB SPL and very broad tuning, consistent with severe IHC and OHC damage. Impaired rate-level slopes were on average shallower than normal for tones, and were steeper in only limited conditions. There was less variation in rate-level slopes across stimuli in impaired fibers, presumably attributable to the lack of suppression-induced reductions in slopes for complex stimuli relative to BF-tone slopes. Sloping saturation was observed less often in impaired fibers. These results illustrate that AN fibers do not provide a simple representation of the basilar-membrane I/O function and suggest that both OHC and IHC damage can affect AN response growth. PMID- 14534290 TI - Spike-frequency adaptation in the inferior colliculus. AB - We investigated spike-frequency adaptation of neurons sensitive to interaural phase disparities (IPDs) in the inferior colliculus (IC) of urethane-anesthetized guinea pigs using a stimulus paradigm designed to exclude the influence of adaptation below the level of binaural integration. The IPD-step stimulus consists of a binaural 3,000-ms tone, in which the first 1,000 ms is held at a neuron's least favorable ("worst") IPD, adapting out monaural components, before being stepped rapidly to a neuron's most favorable ("best") IPD for 300 ms. After some variable interval (1-1,000 ms), IPD is again stepped to the best IPD for 300 ms, before being returned to a neuron's worst IPD for the remainder of the stimulus. Exponential decay functions fitted to the response to best-IPD steps revealed an average adaptation time constant of 52.9 +/- 26.4 ms. Recovery from adaptation to best IPD steps showed an average time constant of 225.5 +/- 210.2 ms. Recovery time constants were not correlated with adaptation time constants. During the recovery period, adaptation to a 2nd best-IPD step followed similar kinetics to adaptation during the 1st best-IPD step. The mean adaptation time constant at stimulus onset (at worst IPD) was 34.8 +/- 19.7 ms, similar to the 38.4 +/- 22.1 ms recorded to contralateral stimulation alone. Individual time constants after stimulus onset were correlated with each other but not with time constants during the best-IPD step. We conclude that such binaurally derived measures of adaptation reflect processes that occur above the level of exclusively monaural pathways, and subsequent to the site of primary binaural interaction. PMID- 14534291 TI - Tyrosine kinase p56lck regulates cell motility and nuclear factor kappaB-mediated secretion of urokinase type plasminogen activator through tyrosine phosphorylation of IkappaBalpha following hypoxia/reoxygenation. AB - Nuclear factor kappaB (NFkappaB) plays major role in regulating cellular responses as a result of environmental injuries. The molecular mechanism(s) by which hypoxia/reoxygenation (H/R) regulates p56lck-dependent activation of NFkappaB through tyrosine phosphorylation of IkappaBalpha and modulates the expression of downstream genes that are involved in cell migration in human breast cancer cells are not well defined. In this paper, we investigated the involvement of protein-tyrosine kinase p56lck in the redox-regulated activation of NFkappaB following H/R in highly invasive (MDA-MB-231) and low invasive (MCF 7) breast cancer cells. We demonstrated that H/R induces tyrosine phosphorylation of p56lck, nuclear translocation of NFkappaB, NFkappaB-DNA binding, and transactivation of NFkappaB through tyrosine phosphorylation of IkappaBalpha. Transfection of these cells with wild type Lck but not with mutant Lck F394 followed by H/R induces the tyrosine phosphorylation of inhibitor of nuclear factor kappaB (IkappaBalpha) and transcriptional activation of NFkappaB, and these are inhibited by Lck inhibitors. In vitro kinase assay demonstrated that immunoprecipitated p56lck but not Lyn or Fyn directly phosphorylate IkappaBalpha in presence of H/R. Pervanadate, H2O2, and H/R induce the interaction between Lck and tyrosine-phosphorylated IkappaBalpha, and this interaction is inhibited by Src homology 2 domain inhibitory peptide, suggesting that tyrosine-phosphorylated IkappaBalpha interacts with Src homology 2 domain of Lck. Luciferase reporter gene assay indicated that Lck induces NFkappaB-dependent urokinase type plasminogen activator (uPA) promoter activity in presence of H/R. Furthermore, H/R stimulates the cell motility through secretion of uPA. To our knowledge, this is the first report that p56lck in presence of H/R regulates NFkappaB activation, uPA secretion, and cell motility through tyrosine phosphorylation of IkappaBalpha and further demonstrates an important redox-regulated pathway for NFkappaB activation following H/R injury that is independent of IkappaB kinase/IkappaBalpha-mediated signaling pathways. PMID- 14534292 TI - Identification of selective inhibitors of NAD+-dependent deacetylases using phenotypic screens in yeast. AB - Sir2 and Hst1 are NAD+-dependent deacetylases involved in transcriptional repression in yeast. The two enzymes are highly homologous yet have different sensitivity to the small-molecule inhibitor splitomicin (compound 1) (Bedalov, A., Gatbonton, T., Irvine, W. P., Gottschling, D. E., and Simon, J. A. (2001) Proc. Natl. Acad. Sci. U. S. A. 98, 15113-15118). We have now defined a critical amino acid residue within a small helical module of Hst1 that confers relative resistance to splitomicin. Parallel cell-based screens of 100 splitomicin analogues led to the identification of compounds that exhibit a higher degree of selectivity toward Sir2 or Hst1. A series of compounds based on a splitomicin derivative, dehydrosplitomicin (compound 2), effectively phenocopied a yeast strain that lacked Hst1 deacetylase while having no effect on the silencing activities of Sir2. In addition, we identified a compound with improved selectivity for Sir2. Selectivity was affirmed using whole-genome DNA microarray analysis. This study underscores the power of phenotypic screens in the development and characterization of selective inhibitors of enzyme functions. PMID- 14534293 TI - Proteomics-based target identification: bengamides as a new class of methionine aminopeptidase inhibitors. AB - LAF389 is a synthetic analogue of bengamides, a class of marine natural products that produce inhibitory effects on tumor growth in vitro and in vivo. A proteomics-based approach has been used to identify signaling pathways affected by bengamides. LAF389 treatment of cells resulted in altered mobility of a subset of proteins on two-dimensional gel electrophoresis. Detailed analysis of one of the proteins, 14-3-3gamma, showed that bengamide treatment resulted in retention of the amino-terminal methionine, suggesting that bengamides directly or indirectly inhibited methionine aminopeptidases (MetAps). Both known MetAps are inhibited by LAF389. Short interfering RNA suppression of MetAp2 also altered amino-terminal processing of 14-3-3gamma. A high resolution structure of human MetAp2 co-crystallized with a bengamide shows that the compound binds in a manner that mimics peptide substrates. Additionally, the structure reveals that three key hydroxyl groups on the inhibitor coordinate the di-cobalt center in the enzyme active site. PMID- 14534294 TI - A bound water molecule is crucial in initiating autocatalytic precursor activation in an N-terminal hydrolase. AB - Cephalosporin acylase is a member of the N-terminal hydrolase family, which is activated from an inactive precursor by autoproteolytic processing to generate a new N-terminal nucleophile Ser or Thr. The gene structure of the precursor cephalosporin acylases generally consists of a signal peptide that is followed by an alpha-subunit, a spacer sequence, and a beta-subunit. The cephalosporin acylase precursor is post-translationally modified into an active heterodimeric enzyme with alpha- and beta-subunits, first by intramolecular cleavage and, second, by intermolecular cleavage. Intramolecular autocatalytic proteolysis is initiated by nucleophilic attack of the residue Ser-1beta onto the adjacent scissile carbonyl carbon. This study determined the precursor structure after disabling the intramolecular cleavage. This study also provides experimental evidence showing that a conserved water molecule plays an important role in assisting the polarization of the OG atom of Ser-1beta to generate a strong nucleophile and to direct the OG atom of the Ser-1beta to a target carbonyl carbon. Intramolecular proteolysis is disabled as a result of a mutation of the residues causing conformational distortion to the active site. This is because distortion affects the existence of the catalytically crucial water at the proper position. This study provides the first evidence showing that a bound water molecule plays a critical role in initiating intramolecular cleavage in the post translational modification of the precursor enzyme. PMID- 14534295 TI - Decreased hepatic triglyceride accumulation and altered fatty acid uptake in mice with deletion of the liver fatty acid-binding protein gene. AB - Liver fatty acid-binding protein (L-Fabp) is an abundant cytosolic lipid-binding protein with broad substrate specificity, expressed in mammalian enterocytes and hepatocytes. We have generated mice with a targeted deletion of the endogenous L Fabp gene and have characterized their response to alterations in hepatic fatty acid flux following prolonged fasting. Chow-fed L-Fabp-/- mice were indistinguishable from wild-type littermates with regard to growth, serum and tissue lipid profiles, and fatty acid distribution within hepatic complex lipid species. In response to 48-h fasting, however, wild-type mice demonstrated a approximately 10-fold increase in hepatic triglyceride content while L-Fabp-/- mice demonstrated only a 2-fold increase. Hepatic VLDL secretion was decreased in L-Fabp-/- mice suggesting that the decreased accumulation of hepatic triglyceride was not the result of increased secretion. Fatty acid oxidation, as inferred from serum beta-hydroxybutyrate levels, was increased in response to fasting, although the increase in L-Fabp-/- mice was significantly reduced in comparison to wild type controls, despite comparable induction of PPAR alpha target genes. Studies in primary hepatocytes revealed indistinguishable initial rates of oleate uptake, but longer intervals revealed reduced rates of uptake in fasted L-Fabp-/- mice. Oleate incorporation into cellular triglyceride and diacylglycerol was reduced in L-Fabp-/- mice although incorporation into phospholipid and cholesterol ester was no different than wild-type controls. These data point to an inducible defect in fatty acid utilization in fasted L-Fabp-/- mice that involves targeting of substrate for use in triglyceride metabolism. PMID- 14534296 TI - Aquifex aeolicus aspartate transcarbamoylase, an enzyme specialized for the efficient utilization of unstable carbamoyl phosphate at elevated temperature. AB - Aquifex aeolicus, an organism that flourishes at 95 degrees C, is one of the most thermophilic eubacteria thus far described. The A. aeolicus pyrB gene encoding aspartate transcarbamoylase (ATCase) was cloned, overexpressed in Escherichia coli, and purified by affinity chromatography to a homogeneous form that could be crystallized. Chemical cross-linking and size exclusion chromatography showed that the protein was a homotrimer of 34-kDa catalytic chains. The activity of A. aeolicus ATCase increased dramatically with increasing temperature due to an increase in kcat with little change in the Km for the substrates, carbamoyl phosphate and aspartate. The Km for both substrates was 30-40-fold lower than the corresponding values for the homologous E. coli ATCase catalytic subunit. Although rapidly degraded at high temperature, the carbamoyl phosphate generated in situ by A. aeolicus carbamoyl phosphate synthetase (CPSase) was channeled to ATCase. The transient time for carbamoyl aspartate formation was 26 s, compared with the much longer transient times observed when A. aeolicus CPSase was coupled to E. coli ATCase. Several other approaches provided strong evidence for channeling and transient complex formation between A. aeolicus ATCase and CPSase. The high affinity for substrates combined with channeling ensures the efficient transfer of carbamoyl phosphate from the active site of CPSase to that of ATCase, thus preserving it from degradation and preventing the formation of toxic cyanate. PMID- 14534297 TI - Crystal structure of a superstable mutant of human p53 core domain. Insights into the mechanism of rescuing oncogenic mutations. AB - Most of the cancer-associated mutations in the tumor suppressor p53 map to its DNA-binding core domain. Many of them inactivate p53 by decreasing its thermodynamic stability. We have previously designed the superstable quadruple mutant M133L/V203A/N239Y/N268D containing the second-site suppressor mutations N239Y and N268D, which specifically restore activity and stability in several oncogenic mutants. Here we present the x-ray structure of this quadruple mutant at 1.9 A resolution, which was solved in a new crystal form in the absence of DNA. This structure reveals that the four point mutations cause only small local structural changes, whereas the overall structure of the central beta-sandwich and the DNA-binding surface is conserved. The suppressor mutation N268D results in an altered hydrogen bond pattern connecting strands S1 and S10, thus bridging the two sheets of the beta-sandwich scaffold in an energetically more favorable way. The second suppressor mutation N239Y, which is located in close proximity to the DNA-binding surface in loop L3, seems to reduce the plasticity of the structure in large parts of loop L3 as indicated by decreased crystallographic temperature factors. The same is observed for residues in the vicinity of the N268D substitution. This increase in rigidity provides the structural basis for the increase in thermostability and an understanding how N268D and N239Y rescue some of the common cancer mutants. PMID- 14534298 TI - Beta-arrestin1 mediates insulin-like growth factor 1 (IGF-1) activation of phosphatidylinositol 3-kinase (PI3K) and anti-apoptosis. AB - beta-arrestins (1 and 2) are widely expressed cytosolic proteins that play central roles in G protein-coupled receptor signaling. beta-arrestin1 is also recruited to the insulin-like growth factor 1 (IGF-1) receptor, a receptor tyrosine kinase, upon agonist binding. Here we report that, in response to IGF-1 stimulation, beta-arrestin1 mediates activation of phosphatidylinositol 3-kinase in a pathway that leads to the subsequent activation of Akt and anti-apoptosis. This process is independent of both Gi and ERK activity. The pathway fails in mouse embryo fibroblasts lacking both beta-arrestins and is restored by stable transfection of beta-arrestin1. Remarkably, this pathway is insensitive to chemical inhibition of IGF-1 receptor tyrosine kinase activity. These results suggest that, in addition to their roles in G protein-coupled receptor signaling, beta-arrestins couple the IGF-1 receptor tyrosine kinase to the phosphatidylinositol 3-kinase system and suggest that this mechanism is operative independently of the tyrosine kinase activity of the receptor. PMID- 14534299 TI - Heterodimerization of type A and B cholecystokinin receptors enhance signaling and promote cell growth. AB - Dimerization of several G protein-coupled receptors has recently been described, but little is known about its clinical and functional relevance. Cholecystokinin (CCK) and gastrin are structurally related gastrointestinal and neuronal peptides whose functions are mediated by two structurally related receptors in this superfamily, the type A and B CCK receptors. We previously demonstrated spontaneous homodimerization of type A CCK receptors and the dissociation of those complexes by agonist occupation (Cheng, Z. J., and Miller, L. J. (2001) J. Biol. Chem. 276, 48040-48047). Here, for the first time, we also demonstrate spontaneous homodimerization of type B CCK receptors, as well as heterodimerization of that receptor with the type A CCK receptor. Unlike type A CCK receptor dimers, the homodimerization of type B CCK receptors was not affected by ligand occupation. However, although heterodimers of type A and B CCK receptors bound natural agonists normally, they exhibited unusual functional and regulatory characteristics. Such complexes demonstrated enhanced agonist stimulated cellular signaling and delayed agonist-induced receptor internalization. As a likely consequence, agonist-stimulated cell growth was markedly enhanced in cells simultaneously expressing both of these receptors. Our results provide the first evidence that heterodimerization of G protein-coupled receptors can form a more "powerful" signaling unit, which has potential clinical significance in promoting cell growth. PMID- 14534300 TI - Multiple states of stalled T7 RNA polymerase at DNA lesions generated by platinum anticancer agents. AB - Transcription inhibition by DNA adducts of cisplatin is considered to be one of the major routes by which this anticancer drug kills cancer cells. Stalled RNA polymerases at platinum-DNA lesions evoke various cellular responses such as nucleotide excision repair, polymerase degradation, and apoptosis. T7 RNA polymerase and site-specifically platinated DNA templates immobilized on a solid support were used to study stalled transcription elongation complexes. In vitro transcription studies were performed in both a promoter-dependent and independent manner. An elongation complex is strongly blocked by cisplatin 1,2 intrastrand d(GpG) and 1,3-intrastrand d(GpTpG) cross-links located on the template strand. Polymerase action is inhibited at multiple sites in the vicinity of the platinum lesion, the nature of which can be altered by the choice and concentration of NTPs. The [(1R,2R-diaminocyclohexane)Pt]2+ DNA adducts formed by oxaliplatin, which carries a stereochemically more demanding spectator ligand than the ammine groups in cisplatin, also strongly block the polymerase with measurable differences compared with cis-[(NH3)2Pt]2+ lesions. Elongation complexes stopped at sites of platinum damage were isolated and characterized. The stalled polymerase can be dissociated from the DNA by subsequent polymerases initiated from the same template. We also discovered that a polymerase stalled at the platinum-DNA lesion can resume transcription after the platinum adduct is chemically removed from the template. PMID- 14534301 TI - Detection of protein folding defects caused by BRCA1-BRCT truncation and missense mutations. AB - Most cancer-associated BRCA1 mutations identified to date result in the premature translational termination of the protein, highlighting a crucial role for the C terminal, BRCT repeat region in mediating BRCA1 tumor suppressor function. However, the molecular and genetic effects of missense mutations that map to the BRCT region remain largely unknown. Using a protease-based assay, we directly assessed the sensitivity of the folding of the BRCT domain to an extensive set of truncation and single amino acid substitutions derived from breast cancer screening programs. The protein can tolerate truncations of up to 8 amino acids, but further deletion results in drastic BRCT folding defects. This molecular phenotype can be correlated with an increased susceptibility to disease. A cross validated computational assessment of the BRCT mutation data base suggests that as much as half of all BRCT missense mutations contribute to BRCA1 loss of function and disease through protein-destabilizing effects. The coupled use of proteolytic methods and computational predictive methods to detect mutant BRCA1 conformations at the protein level will augment the efficacy of current BRCA1 screening protocols, especially in the absence of clinical data that can be used to discriminate deleterious BRCT missense mutations from benign polymorphisms. PMID- 14534302 TI - Modulation of glucocorticoid receptor transcriptional activation, phosphorylation, and growth inhibition by p27Kip1. AB - The cyclin-dependent kinase inhibitor p27Kip1 is frequently inactivated in human cancers. Glucocorticoids, acting through the glucocorticoid receptor (GR), are frequently used to treat certain malignancies and are growth inhibitive, but the relationship between GR activity and p27 status has not been explored. We have therefore examined GR-dependent transcriptional activation, receptor phosphorylation, and glucocorticoid-dependent growth inhibition in p27-deficient (p27-/-) murine embryonic fibroblasts (MEFs). We find that GR transcriptional enhancement as well as receptor phosphorylation at two putative cyclin-dependent kinase sites are elevated in p27-/- MEFs, relative to control cells. This increased GR transcriptional activation appears to be mediated through the GR N terminus, and coexpression of the GR N-terminal coactivator, DRIP150, further enhanced GR-dependent transcriptional activation. Furthermore, p27-/- MEFs are partially resistant to the growth inhibitory effects of glucocorticoids. Thus, p27 appears to be an important element in the GR transcription and growth inhibitory responses. PMID- 14534303 TI - Reversal of cyanide inhibition of cytochrome c oxidase by the auxiliary substrate nitric oxide: an endogenous antidote to cyanide poisoning? AB - Nitric oxide (NO) is shown to overcome the cyanide inhibition of cytochrome c oxidase in the presence of excess ferrocytochrome c and oxygen. Addition of NO to the partially reduced cyanide-inhibited form of the bovine enzyme is shown by electron paramagnetic resonance spectroscopy to result in substitution of cyanide at ferriheme a3 by NO with reduction of the heme. The resulting nitrosylferroheme a3 is a 5-coordinate structure, the proximal bond to histidine having been broken. NO does not simply act as a reversibly bound competitive inhibitor but is an auxiliary substrate consumed in a catalytic cycle along with ferrocytochrome c and oxygen. The implications of this observation with regard to estimates of steady-state NO levels in vivo is discussed. Given the multiple sources of NO available to mitochondria, the present results appear to explain in part some of the curious biomedical observations reported by other laboratories; for example, the kidneys of cyanide poisoning victims surprisingly exhibit no significant irreversible damage, and lethal doses of potassium cyanide are able to inhibit cytochrome c oxidase activity by only approximately 50% in brain mitochondria. PMID- 14534304 TI - Metalloproteinase expression in PMA-stimulated THP-1 cells. Effects of peroxisome proliferator-activated receptor-gamma (PPAR gamma) agonists and 9-cis-retinoic acid. AB - The PPAR gamma agonists, thiazolidinediones (TZDs), have anti-inflammatory properties as well as increasing insulin sensitivity. This has widened their therapeutic scope to treat inflammatory diseases such as atherosclerosis in addition to Type 2 Diabetes. TZDs are known to reduce monocyte/macrophage expression of Matrix metalloproteinase (MMP)-9, which is implicated in atherosclerotic plaque destabilization. This study aims to identify other metalloproteinase genes of the ADAM (A Disintegin And Metalloproteinase) and ADAMTS families that are regulated by PPAR gamma or RXR agonists, which are potentially important in type 2 diabetes and/or related atherosclerosis. The synthetic PPAR gamma agonist, GW7845, and the natural agonist 15d-PGJ2, suppressed PMA stimulated MMP-9 in human monocyte-like cells (THP-1) only in the presence of 9-cis-retinoic acid. Quantitative Real-Time PCR showed that this reduction was regulated at the mRNA level. Expression of ADAMs 8, 9, and 17 were increased, and ADAM15 was decreased by stimulation of THP-1 with PMA, although these ADAMs were not regulated by PPAR gamma or RXR agonists. PMA-induced ADAM28 expression was further enhanced by the addition of 9-cis-retinoic acid. ADAMTS4, implicated in rheumatoid arthritis, was expressed in THP-1 cells, and significantly increased after 24 h of PMA stimulation. ADAMTS4 expression was suppressed by both PPAR gamma and RXR agonists and was undetectable when the agonists were combined. Pretreatment of THP-1 cells with the PPAR gamma antagonist, GW9662, suggests that PPAR gamma plays subtly different roles in the regulation of MMP-9, ADAMTS4 and ADAM28 gene expression. These results indicate that PPAR gamma and RXR agonists have complex effects on monocyte metalloproteinase expression, which may have implications for therapeutic strategies. PMID- 14534305 TI - Store-operated Ca2+ current and TRPV6 channels in lymph node prostate cancer cells. AB - The contribution of endogenous and recombinant transient receptor potential vanilloid type 6 (TRPV6) channels to Ca2+ entry across the plasma membrane was studied in the human lymph node prostate cancer cell line (LNCaP). LNCaP cells do express the TRPV6 gene, and Ca2+ entry currents in these cells were detected after active and passive Ca2+ store depletion by intracellular application of inositol 1,4,5-trisphosphate, Ca2+ chelators, and the sarcoplasmic/endoplasmic reticulum Ca2+-ATPase inhibitor thapsigargin. This store-operated Ca2+ current (ISOC) had biophysical properties similar to those of the Ca2+ release-activated Ca2+ current (ICRAC) in rat basophilic leukemia cells such as the activation mechanism, inward rectification, and Ca2+ selectivity. These properties are also shared by the Ca2+-sensing Ca2+ current (ITRPV6) recorded after heterologous expression of TRPV6 cDNA in human embryonic kidney and rat basophilic leukemia cells (Bodding, M., Wissenbach, U., Flockerzi, V. (2002) J. Biol. Chem. 277, 36656-36664). TRPV6 cDNA transfection of LNCaP cells restored recombinant ITRPV6, which can be distinguished from ISOC by the mechanism of activation, the voltage dependence of monovalent currents in the absence of external divalent cations, and the changes in Ca2+ current densities due to different membrane potentials. In addition, ISOC was not affected by antiandrogen or 1,25-dihydroxyvitamin D3 treatment of LNCaP cells, which up-regulates TRPV6 gene expression, or by androgen treatment, which has the opposite effect. Therefore, native channels responsible for ISOC are different from those for recombinant ITRPV6 and do not appear to be affected if one of their assumed subunits, TRPV6, is up- or down regulated, suggesting a rather rigid subunit composition in vivo. PMID- 14534306 TI - Exploratory and confirmatory gene expression profiling of mac1Delta. AB - Exploratory outlier identification methods and confirmatory gene expression studies showed induction of the iron regulon in Saccharomyces cerevisiae lacking Mac1p, a copper-responsive transcription factor. The Aft1p/Aft2p binding motif was the most discriminating motif between up- and down-regulated genes, and we identified new genes potentially regulated by Aft1p/Aft2p. In addition, multiple genes encoding proteins containing Fe-S clusters were down-regulated suggesting metabolic reorganization to conserve iron in mac1Delta. Null mutants of each of the differentially expressed genes were characterized for copper- or iron-related phenotypes. New or additional support for a role in copper and iron homeostasis is provided in this study for the gene products of AKR1, MRS4, PCA1, SSU1, TIS11, YBR047W, YHL035C, YHR045W, YLR047C, YLR126C, and YTP1. PMID- 14534307 TI - The N-terminal input domain of the sensor kinase KdpD of Escherichia coli stabilizes the interaction between the cognate response regulator KdpE and the corresponding DNA-binding site. AB - The sensor kinase/response regulator system KdpD/KdpE of Escherichia coli regulates expression of the kdpFABC operon, which encodes the high affinity K+ transport system KdpFABC. The membrane-bound sensor kinase KdpD consists of an N terminal input domain (comprising a large cytoplasmic domain and four transmembrane domains) and a cytoplasmic C-terminal transmitter domain. Here we show that the cytoplasmic N-terminal domain of KdpD (KdpD/1-395) alone supports semi-constitutive kdpFABC expression, which becomes dependent on the extracellular K+ concentration under K+-limiting growth conditions. However, it should be noted that the non-phosphorylatable derivative KdpD/H673Q or the absence of KdpD abolishes kdpFABC expression completely. KdpD/1-395 mediated kdpFABC expression requires the corresponding response regulator KdpE with an intact phosphorylation site. Experiments with an Escherichia coli mutant unable to synthesize acetyl phosphate as well as transposon mutagenesis suggest that KdpE is phosphorylated in vivo by low molecular weight phosphodonors in the absence of the full-length sensor kinase. Various biochemical approaches provide first evidence that kdpFABC expression mediated by KdpD/1-395 is due to a stabilizing effect of this domain on the binding of KdpE approximately P to its corresponding DNA-binding site. Such a stabilizing effect of a sensor kinase domain on the DNA-protein interaction of the cognate response regulator has never been observed before for any other sensor kinase. It describes a new mechanism in bacterial two-component signal transduction. PMID- 14534308 TI - Quantitative analysis of platelet alpha v beta 3 binding to osteopontin using laser tweezers. AB - To determine whether platelet adhesion to surfaces coated with the matrix protein osteopontin requires an agonist-induced increase in the affinity of the integrin alpha v beta 3 for this ligand, we used laser tweezers to measure the rupture force between single alpha v beta 3 molecules on the platelet surface and osteopontin-coated beads. Virtually all platelets stimulated with 10 microM ADP bound strongly to osteopontin, producing rupture forces as great as 100 piconewtons (pN) with a peak at 45-50 pN. By contrast, 90% of unstimulated, resting non-reactive platelets bound weakly to osteopontin, with rupture forces rarely exceeding 30-35 pN. However, approximately 10% of unstimulated platelets, resting reactive platelets, exhibited rupture force distributions similar to stimulated platelets. Moreover, ADP stimulation resulted in a 12-fold increase in the probability of detecting rupture forces >30 pN compared with resting non reactive platelets. Pre-incubating stimulated platelets with the inhibitory prostaglandin E1, a cyclic RGD peptide, the monoclonal antibody abciximab, or the alpha v beta 3-specific cyclic peptide XJ735 returned force histograms to those of non-reactive platelets. These experiments demonstrate that ADP stimulation increases the strength of the interaction between platelet alpha v beta 3 and osteopontin. Furthermore, they indicate that platelet adhesion to osteopontin coated surfaces requires an agonist-induced exposure of alpha v beta 3-binding sites for this ligand. PMID- 14534309 TI - Elevated intracellular calcium triggers recruitment of the receptor cross-talk accessory protein calcyon to the plasma membrane. AB - Calcyon is called a "cross-talk accessory protein" because the mechanism by which it enables the typically Gs-linked D1 dopamine receptor to stimulate intracellular calcium release depends on a priming step involving heterologous Gq linked G-protein-coupled receptor activation. The details of how priming facilitates the D1R calcium response have yet to be precisely elucidated. The present work shows that calcyon is constitutively localized both in vesicular and plasma membrane compartments within HEK293 cells. In addition, surface biotinylation and luminescence assays revealed that priming stimulates a 2-fold increase in the levels of calcyon expressed on the cell surface and that subsequent D1R activation produces further accumulation of the protein in the plasma membrane. The effects of priming and D1R agonists were blocked by nocodazole implicating microtubules in the delivery of calcyon-containing vesicles to the cell surface. Accumulation of calcyon in the plasma membrane correlated well with increased intracellular calcium levels as thapsigargin mimicked, and 2-aminoethoxydiphenylborane abrogated, the effects of priming. KN 62, an inhibitor of calcium/calmodulin-dependent protein kinase II (CaMKII) also blocked the effects of priming and D1R agonists. Furthermore, expression of constitutively active forms of the kinase bypassed the requirement for priming indicating that CaMKII is a key effector in the Ca2+ and microtubule-dependent delivery of calcyon to the cell surface. PMID- 14534310 TI - Structures of gamma-aminobutyric acid (GABA) aminotransferase, a pyridoxal 5' phosphate, and [2Fe-2S] cluster-containing enzyme, complexed with gamma-ethynyl GABA and with the antiepilepsy drug vigabatrin. AB - Gamma-aminobutyric acid aminotransferase (GABA-AT) is a pyridoxal 5'-phosphate dependent enzyme responsible for the degradation of the inhibitory neurotransmitter GABA. GABA-AT is a validated target for antiepilepsy drugs because its selective inhibition raises GABA concentrations in brain. The antiepilepsy drug, gamma-vinyl-GABA (vigabatrin) has been investigated in the past by various biochemical methods and resulted in several proposals for its mechanisms of inactivation. In this study we solved and compared the crystal structures of pig liver GABA-AT in its native form (to 2.3-A resolution) and in complex with vigabatrin as well as with the close analogue gamma-ethynyl-GABA (to 2.3 and 2.8 A, respectively). Both inactivators form a covalent ternary adduct with the active site Lys-329 and the pyridoxal 5'-phosphate (PLP) cofactor. The crystal structures provide direct support for specific inactivation mechanisms proposed earlier on the basis of radio-labeling experiments. The reactivity of GABA-AT crystals with the two GABA analogues was also investigated by polarized absorption microspectrophotometry. The spectral data are discussed in relation to the proposed mechanism. Intriguingly, all three structures revealed a [2Fe-2S] cluster of yet unknown function at the center of the dimeric molecule in the vicinity of the PLP cofactors. PMID- 14534311 TI - Bcl-XL mutations suppress cellular sensitivity to antimycin A. AB - Cells expressing high levels of the BCL-X(L) anti-apoptotic protein are preferentially killed by the mitochondrial inhibitor antimycin A (AA). Computational modeling predicts a binding site for AA in the extended hydrophobic groove on BCL-X(L), previously identified as an interface for dimerization to BAX and related proapoptotic proteins. Here, we identify BCL-X(L) hydrophobic groove mutants with normal cellular anti-apoptotic function but suppressed sensitivity to AA. The LD(50) of AA for cells expressing BCL-X(L) mutants directly correlates with the measured in vitro dissociation constants for AA binding. These results indicate that BCL-X(L) is a principal target mediating AA cytotoxicity. PMID- 14534312 TI - Understanding resistance to beta-lactams and beta-lactamase inhibitors in the SHV beta-lactamase: lessons from the mutagenesis of SER-130. AB - Bacterial resistance to beta-lactam/beta-lactamase inhibitor combinations by single amino acid mutations in class A beta-lactamases threatens our most potent clinical antibiotics. In TEM-1 and SHV-1, the common class A beta-lactamases, alterations at Ser-130 confer resistance to inactivation by the beta-lactamase inhibitors, clavulanic acid, and tazobactam. By using site-saturation mutagenesis, we sought to determine the amino acid substitutions at Ser-130 in SHV-1 beta-lactamase that result in resistance to these inhibitors. Antibiotic susceptibility testing revealed that ampicillin and ampicillin/clavulanic acid resistance was observed only for the S130G beta-lactamase expressed in Escherichia coli. Kinetic analysis of the S130G beta-lactamase demonstrated a significant elevation in apparent Km and a reduction in kcat/Km for ampicillin. Marked increases in the dissociation constant for the preacylation complex, KI, of clavulanic acid (SHV-1, 0.14 microm; S130G, 46.5 microm) and tazobactam (SHV 1, 0.07 microm; S130G, 4.2 microm) were observed. In contrast, the k(inact)s of S130G and SHV-1 differed by only 17% for clavulanic acid and 40% for tazobactam. Progressive inactivation studies showed that the inhibitor to enzyme ratios required to inactivate SHV-1 and S130G were similar. Our observations demonstrate that enzymatic activity is preserved despite amino acid substitutions that significantly alter the apparent affinity of the active site for beta-lactams and beta-lactamase inhibitors. These results underscore the mechanistic versatility of class A beta-lactamases and have implications for the design of novel beta lactamase inhibitors. PMID- 14534313 TI - Surface-exposed glycopeptidolipids of Mycobacterium smegmatis specifically inhibit the phagocytosis of mycobacteria by human macrophages. Identification of a novel family of glycopeptidolipids. AB - Phagocytosis by macrophages represents the early step of the mycobacterial infection. It is governed both by the nature of the host receptors used and the ligands exposed on the bacteria. The outermost molecules of the nonpathogenic Mycobacterium smegmatis were extracted by a mechanical treatment and found to specifically and dose dependently inhibit the phagocytosis of both M. smegmatis and the opportunistic pathogen M. kansasii by human macrophages derived from monocytes. The inhibitory activity was attributed to surface lipids because it is extracted by chloroform and reduced by alkaline hydrolysis but not by protease treatment. Fractionation of surface lipids by adsorption chromatography indicated that the major inhibitory compounds consisted of phospholipids and glycopeptidolipids (GPLs). Mass spectrometry and nuclear magnetic resonance spectroscopy analyses, combined with chemical degradation methods, demonstrated the existence of a novel family of GPLs that consists of a core composed of the long-chain tripeptidyl amino-alcohol with a di-O-acetyl-6-deoxytalosyl unit substituting the allo-threoninyl residue and a 2-succinyl-3,4-di-O-CH3-rhamnosyl unit linked to the alaninol end of the molecules. These compounds, as well as diglycosylated GPLs at the alaninol end and de-O-acylated GPLs, but not the non serovar-specific di-O-acetylated GPLs, inhibited the phagocytosis of M. smegmatis and M. avium by human macrophages at a few nanomolar concentration without affecting the rate of zymosan internalization. At micromolar concentrations, the native GPLs also inhibit the uptake of both M. tuberculosis and M. kansasii. De-O acylation experiments established the critical roles of both the succinyl and acetyl substituents. Collectively, these data provide evidence that surface exposed mycobacterial glycoconjugates are efficient competitors of the interaction between macrophages and mycobacteria and, as such, could represent pharmacological tools for the control of mycobacterial infections. PMID- 14534314 TI - Mediator influences Schizosaccharomyces pombe RNA polymerase II-dependent transcription in vitro. AB - The fission yeast Schizosaccharomyces pombe has proved an important model system for cross-species comparative studies of many fundamental processes in the eukaryotic cell, such as cell cycle control and DNA replication. The RNA polymerase II transcription machinery is, however, still relatively poorly understood in S. pombe, partially due to the absence of a reconstituted in vitro transcription system. We have now purified S. pombe RNA polymerase II and its general initiation factors TFIIB, TFIIF, TFIIE, and TFIIH to near homogeneity. These factors enable RNA polymerase II to initiate transcription from the S. pombe alcohol dehydrogenase promoter (adh1p) when combined with Saccharomyces cerevisiae TATA-binding protein. We use our reconstituted system to examine effects of Mediator on basal transcription in vitro. S. pombe Mediator exists in two distinct forms, a free form, which contains the spSrb8, spTrap240, spSrb10, and spSrb11 subunits, and a smaller form, which lacks these four subunits and associates with RNA polymerase II to form a holoenzyme. We find that spSrb8/spTrap240/spSrb10/spSrb11 containing Mediator repress basal transcription, whereas Mediator lacking these subunits has a stimulatory effect on transcription. Our findings thus demonstrate that the spSrb8/spTrap240/spSrb10/spSrb11 subcomplex governs the ability of Mediator to stimulate or repress basal transcription in vitro. PMID- 14534315 TI - Identification and characterization of an intermediate in the alkali degradation of (6-4) photoproduct-containing DNA. AB - The (6-4) photoproduct formed by ultraviolet light is known as an alkali-labile DNA lesion. Strand breaks occur at (6-4) photoproducts when UV-irradiated DNA is treated with hot alkali. We have analyzed the degradation reaction of this photoproduct under alkaline conditions using synthetic oligonucleotides. A tetramer, d(GT(6-4)TC), was prepared, and its degradation in 50 mm KOH at 60 degrees C was monitored by high performance liquid chromatography. A single peak with a UV absorption spectrum similar to that of the starting material was detected after the reaction, and this compound was regarded as an intermediate before the strand break. The formation of this intermediate was compared with intermediates from the degradation of other alkali-labile lesions such as the abasic site, thymine glycol, and 5,6-dihydrothymine. The results strongly suggested that the first step of the alkali degradation of the (6-4) photoproduct was the hydrolysis between the N3 and C4 positions of the 5'-pyrimidine component. Analyses by NMR spectroscopy and mass spectrometry supported the chemical structure of this product. Assays of the complex formation with XPC.HR23B and the translesion synthesis by DNA polymerase eta revealed that the biochemical properties are indistinguishable between the intact and hydrolyzed photoproducts. PMID- 14534316 TI - A cytochrome b562 variant with a c-type cytochrome CXXCH heme-binding motif as a probe of the Escherichia coli cytochrome c maturation system. AB - Cytochrome b562 is a periplasmic Escherichia coli protein; previous work has shown that heme can be attached covalently in vivo as a consequence of introduction of one or two cysteines into the heme-binding pocket. A heterogeneous mixture of products was obtained, and it was not established whether the covalent bond formation was catalyzed or spontaneous. Here, we show that coexpression from plasmids of a variant of cytochrome b562 containing a CXXCH heme-binding motif with the E. coli cytochrome c maturation (Ccm) proteins results in an essentially homogeneous product that is a correctly matured c-type cytochrome. Formation of the holocytochrome was accompanied by substantial production of its apo form, in which, for the protein as isolated, there is a disulfide bond between the two cysteines in the CXXCH motif. Following addition of heme to reduced CXXCH apoprotein, spontaneous covalent addition of heme to polypeptide occurred in vitro. Strikingly, the spectral properties were very similar to those of the material obtained from cells in which presumed uncatalyzed addition of heme (i.e. in the absence of Ccm) had been observed. The major product from uncatalyzed heme attachment was an incorrectly matured cytochrome with the heme rotated by 180 degrees relative to its normal orientation. The contrast between Ccm-dependent and Ccm-independent covalent attachment of heme indicates that the Ccm apparatus presents heme to the protein only in the orientation that results in formation of the correct product and also that heme does not become covalently attached to the apocytochrome b562 CXXCH variant without being handled by the Ccm system in the periplasm. The CXXCH variant of cytochrome b562 was also expressed in E. coli strains deficient in the periplasmic reductant DsbD or oxidant DsbA. In the DsbA- strain under aerobic conditions, c-type cytochromes were made abundantly and correctly when the Ccm proteins were expressed. This contrasts with previous reports indicating that DsbA is essential for cytochrome c biogenesis in E. coli. PMID- 14534317 TI - Characterization of HdnoR, the transcriptional repressor of the 6-hydroxy-D nicotine oxidase gene of Arthrobacter nicotinovorans pAO1, and its DNA-binding activity in response to L- and D-nicotine Derivatives. AB - Utilization of L-nicotine as growth substrate by Arthrobacter nicotinovorans pAO1 starts with hydroxylation of the pyridine ring at C6. Next, the pyrrolidine ring is oxidized by 6-hydroxy-L-nicotine oxidase, which acts strictly stereo-specific on the L-enantiomer. Surprisingly, L-nicotine also induces the synthesis of a 6 hydroxy-d-nicotine-specific oxidase in the bacteria. Genes of nicotine-degrading enzymes are located on the catabolic plasmid pAO1. The pAO1 sequence revealed that the 6-hydroxy-D-nicotine oxidase gene is flanked by two open reading frames with a similarity to amino acid permeases and a divergently transcribed open reading frame with a similarity to proteins of the tetracycline repressor TetR family. Reverse transcription PCR and primer extension analysis of RNA transcripts isolated from A. nicotinovorans pAO1 indicated that the 6-hydroxy-D nicotine oxidase gene represents a transcriptional unit. DNA electromobility shift assays established that the purified TetR-similar protein represents the 6 hydroxy-D-nicotine oxidase gene repressor HdnoR and binds to the 6-hydroxy-D nicotine oxidase gene operator with a Kd of 21 nM. The enantiomers 6-hydroxy-D- and 6-hydroxy-L-nicotine acted in vitro as inducers. In vivo analysis of 6 hydroxy-D-nicotine oxidase gene transcripts from bacteria grown with L- and D nicotine confirmed this conclusion. The poor discrimination by HdnoR between the 6-hydroxy-L- and 6-hydroxy-D-nicotine enantiomers explains the presence of the 6 hydroxy-D-nicotine-specific enzyme in bacteria grown on L-nicotine. PMID- 14534318 TI - Self-association of the amino-terminal domain of the yeast TATA-binding protein. AB - The amino-terminal domain of yeast TATA-binding protein has been proposed to play a crucial role in the self-association mechanism(s) of the full-length protein. Here we tested the ability of this domain to self-associate under a variety of solution conditions. Escherichia coli two-hybrid assays, in vitro pull-down assays, and in vitro cross-linking provided qualitative evidence for a limited and specific self-association. Sedimentation equilibrium analysis using purified protein was consistent with a monomer-dimer equilibrium with an apparent dissociation constant of approximately 8.4 microM. Higher stoichiometry associations remain possible but could not be detected by any of these methods. These results demonstrate that the minimal structure necessary for amino-terminal domain self-association must be present even in the absence of carboxyl-terminal domain structures. On the basis of these results we propose that amino-terminal domain structures contribute to the oligomerization interface of the full-length yeast TATA-binding protein. PMID- 14534319 TI - Overexpression of repressive cAMP response element modulators in high glucose and fatty acid-treated rat islets. A common mechanism for glucose toxicity and lipotoxicity? AB - The hyperlipidemia and hyperglycemia of the diabetic state accelerate beta-cell dysfunction, yet the mechanisms are not fully defined. We used rat islet-specific oligonucleotide arrays (Metabolex Rat Islet Genechips) to identify genes that are coordinately regulated by high glucose and free fatty acids (FFA). Exposure of rat islets to FFA (125 microM for 2 days) or glucose (27 mM for 4 days) reduced glucose-stimulated insulin secretion by 70 +/- 5 and 40 +/- 4%, respectively, relative to control-cultured islets. These treatments also substantially reduced the insulin content of the islets. Islet Genechips analysis revealed that the mRNA levels of cAMP response element modulator (CREM)-17X and inducible cAMP early repressor were significantly increased in both 27 mM glucose- and FFA treated islets. Removing FFA or high glucose from the culture medium restored glucose-stimulated insulin secretion and the mRNA levels of the two CREM repressors to normal. Northern blot analysis revealed a 5-fold increase in the abundance of CREM-17X mRNA and a concomitant 50% reduction in the insulin mRNA in FFA-treated islets. Transient transfection of the insulin-secreting beta HC9 cells with CREM-17X suppressed rat insulin promoter activity by nearly 50%. Overexpression of CREM-17X in intact islets via adenovirus infection decreased islet insulin mRNA levels and insulin content and resulted in a significant decrease in glucose- or KCl-induced insulin secretion. Taken together, these data suggest that up-regulation of CREM repressors by either FFA or high glucose exacerbates beta-cell failure in type 2 diabetes by suppressing insulin gene transcription. PMID- 14534320 TI - Werner syndrome protein contains three structure-specific DNA binding domains. AB - Werner syndrome (WS) is a premature aging syndrome caused by mutations in the WS gene (WRN) and a deficiency in the function of the Werner protein (WRN). WRN is a multifunctional nuclear protein that catalyzes three DNA-dependent reactions: a 3'-5'-exonuclease, an ATPase, and a 3'-5'-helicase. Deficiency in WRN results in a cellular phenotype of genomic instability. The biochemical characteristics of WRN and the cellular phenotype of WRN mutants suggest that WRN plays an important role in DNA metabolic pathways such as recombination, transcription, replication, and repair. The catalytic activities of WRN have been extensively studied and are fairly well understood. However, much less is known about the domain-specific interactions between WRN and its DNA substrates. This study identifies and characterizes three distinct WRN DNA binding domains using recombinant truncated fragments of WRN and five DNA substrates (long forked duplex, blunt-ended duplex, single-stranded DNA, 5'-overhang duplex, and Holliday junction). Substrate specific DNA binding activity was detected in three domains, one N-terminal and two different C-terminal WRN fragments (RecQ conserved domain and helicase RNase D conserved domain-containing domains). The substrate specificity of each DNA binding domain may indicate that each protein domain has a distinct biological function. The importance of these results is discussed with respect to proposed roles for WRN in distinct DNA metabolic pathways. PMID- 14534325 TI - The jasmonate biochemical pathway. AB - Plants possess an interrelated and interacting family of potent fatty acid derived regulators--the jasmonates. These compounds, which play roles in both defense and development, are derived from tri-unsaturated fatty acids [alpha linolenic acid (18:3) or 7Z,10Z,13Z-hexadecatrienoic acid (16:3)]. The lipoxygenase-catalyzed addition of molecular oxygen to alpha-linolenic acid initiates jasmonate synthesis by providing a 13-hydroperoxide substrate for the formation of an unstable allene oxide that is then subject to enzyme-guided cyclization to produce 12-oxo-phytodienoic acid (OPDA). OPDA, a key regulatory lipid in the plant immune system, has several fates, including esterification into plastid lipids or transformation into the 12-carbon co-regulator jasmonic acid (JA). JA, the best-characterized member of the family, regulates both male and female fertility (depending on the plant species), and is an important mediator of defense gene expresssion. JA is itself a substrate for further diverse modifications. Genetic dissection of the pathway is revealing how the different jasmonates modulate different physiological processes. Each new family member that is discovered provides another key to understanding the fine control of gene expression in immune responses, in the initiation and maintenance of long distance signal transfer in response to wounding, and in the regulation of fertility, among other processes. The Jasmonate Biochemical Pathway provides an overview of the growing jasmonate family, and new members will be included in future versions of the Connections Map. Science Viewpoint R. Liechti, E. E. Farmer, The jasmonate pathway. Science 296, 1649-1650 (2002). [Abstract] [Full Text] PMID- 14534321 TI - Yeast beta-alanine synthase shares a structural scaffold and origin with dizinc dependent exopeptidases. AB - beta-Alanine synthase (beta AS) is the final enzyme of the reductive pyrimidine catabolic pathway, which is responsible for the breakdown of pyrimidine bases, including several anticancer drugs. In eukaryotes, beta ASs belong to two subfamilies, which exhibit a low degree of sequence similarity. We determined the structure of beta AS from Saccharomyces kluyveri to a resolution of 2.7 A. The subunit of the homodimeric enzyme consists of two domains: a larger catalytic domain with a dizinc metal center, which represents the active site of beta AS, and a smaller domain mediating the majority of the intersubunit contacts. Both domains exhibit a mixed alpha/beta-topology. Surprisingly, the observed high structural homology to a family of dizinc-dependent exopeptidases suggests that these two enzyme groups have a common origin. Alterations in the ligand composition of the metal-binding site can be explained as adjustments to the catalysis of a different reaction, the hydrolysis of an N-carbamyl bond by beta AS compared with the hydrolysis of a peptide bond by exopeptidases. In contrast, there is no resemblance to the three-dimensional structure of the functionally closely related N-carbamyl-d-amino acid amidohydrolases. Based on comparative structural analysis and observed deviations in the backbone conformations of the eight copies of the subunit in the asymmetric unit, we suggest that conformational changes occur during each catalytic cycle. PMID- 14534326 TI - Finally, proof of weapons of mass destruction. AB - Allelopathy (one species' use of chemicals to harm other species) may be a key ingredient in successful invasions of alien plants into established communities. Bais et al. show that in response to elicitation by common soil fungi, spotted knapweed (Centaurea maculosa) launches an ineffective defense against the fungi that results in extensive collateral damage to neighboring plants. Specifically, the flavonoid (-)-catechin, released from the roots of knapweed, produces a massive reactive oxygen species (ROS)-mediated oxidative burst, Ca2+ signaling, and rapid cell death in those unadapted native species whose habitats the plant regularly invades. The roots release both (+) and (-) enantiomers, but only the ( ) enantiomer functions as a weapon of mass destruction; the (+) enantiomer inhibits the growth of numerous common soil-borne bacterial pathogens. Eliciting apoptotic response for a competitive advantage is an example of signal cross-talk between the genomes of interacting organisms and highlights how the internal signaling of one organism can be used by others to adjust their phenotypes in an adaptive manner. The study provides strong circumstantial evidence for an allelopathic interaction, but the genetic manipulation of (-)-catechin release would allow researchers to determine if these responses occur in nature. Precise genetic control over the release of secondary metabolites from plants would benefit ecological research. PMID- 14534327 TI - Reducing integrins improves the quality of fly life. AB - Integrins are cell surface receptors that mediate adhesion to the extracellular matrix. In this Perspective, we focus on the recent finding that a decrease in the levels of integrins yields fruit flies that retain youthful mobility as they age and display an increased mean life span. We discuss how the reduction of integrin activity might lead to the observed changes. PMID- 14534328 TI - mTor is required for hypertrophy of Pten-deficient neuronal soma in vivo. AB - The mechanisms that regulate mammalian cell size during development and homeostatic maintenance are poorly understood. The tumor suppressor Pten is required for correct maintenance of mammalian neuronal soma size. Selective inactivation of Pten in postnatal granule neurons of the cerebellum and dentate gyrus in mouse causes cell-autonomous hypertrophy as well as more complex phenotypes, including progressive macrocephaly, seizures, and premature death. To determine the contribution of mTor signaling to Pten-mediated growth regulation in the mammalian nervous system, we treated Pten conditional knockout mice with CCI-779, a specific mTor inhibitor. mTor inhibition decreased the seizure frequency and death rate in Pten mutant mice, prevented the increase in Pten deficient neuronal soma size in young mice, and reversed neuronal soma enlargement in adult mice. mTor inhibition did not decrease the size of wild-type adult neurons. Thus, mTor is required for neuronal hypertrophy downstream of Pten deficiency, but is not required for maintenance of normal neuronal soma size. mTOR inhibitors may be useful therapeutic agents for diseases in brain resulting from PTEN deficiency such as Lhermitte-Duclos disease or glioblastoma multiforme. PMID- 14534329 TI - Cyclothiazide potently inhibits gamma-aminobutyric acid type A receptors in addition to enhancing glutamate responses. AB - Ionotropic glutamate and gamma-aminobutyric acid type A (GABAA) receptors mediate critical excitatory and inhibitory actions in the brain. Cyclothiazide (CTZ) is well known for its effect of enhancing glutamatergic transmission and is widely used as a blocker for alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptor desensitization. Here, we report that in addition to its action on AMPA receptors, CTZ also exerts a powerful but opposite effect on GABAA receptors. We found that CTZ reversibly inhibited both evoked and spontaneous inhibitory postsynaptic currents, as well as GABA application-induced membrane currents, in a dose-dependent manner. Single-channel analyses revealed further that CTZ greatly reduced the open probability of GABAA receptor channels. These results demonstrate that CTZ interacts with both glutamate and GABAA receptors and shifts the excitation-inhibition balance in the brain by two independent mechanisms. Understanding the molecular mechanism of this double faceted drug-receptor interaction may help in designing new therapies for neurological diseases. PMID- 14534330 TI - An endogenous retroviral long terminal repeat is the dominant promoter for human beta1,3-galactosyltransferase 5 in the colon. AB - LTRs of endogenous retroviruses are known to affect expression of several human genes, typically as a relatively minor alternative promoter. Here, we report that an endogenous retrovirus LTR acts as one of at least two alternative promoters for the human beta1,3-galactosyltransferase 5 gene, involved in type 1 Lewis antigen synthesis, and show that the LTR promoter is most active in the gastrointestinal tract and mammary gland. Indeed, the LTR is the dominant promoter in the colon, indicating that this ancient retroviral element has a major impact on gene expression. Using colorectal cancer cell lines and electrophoretic mobility-shift assays, we found that hepatocyte nuclear factor 1 (HNF-1) binds a site within the retroviral promoter and that expression of HNF-1 and interaction with its binding site correlated with promoter activation. We conclude that HNF-1 is at least partially responsible for the tissue-specific activation of the LTR promoter of human beta 1,3-galactosyltransferase 5. We demonstrate that this tissue-specific transcription factor is implicated in the activation of an LTR gene promoter. PMID- 14534331 TI - Minimal residual disease in chronic myeloid leukemia. PMID- 14534332 TI - Pharmacologic approaches to correcting the basic defect in cystic fibrosis. PMID- 14534333 TI - First-trimester screening for trisomies 21 and 18. AB - BACKGROUND: Screening for aneuploid pregnancies is routinely performed after 15 weeks of gestation and has a sensitivity of approximately 65 percent, with a false positive rate of 5 percent. First-trimester markers of aneuploidy have been developed, but their use in combination has not been adequately evaluated in clinical practice. METHODS: We conducted a multicenter study of screening for trisomies 21 and 18 among patients with pregnancies between 74 and 97 days of gestation, based on maternal age, maternal levels of free beta human chorionic gonadotropin and pregnancy-associated plasma protein A, and ultrasonographic measurement of fetal nuchal translucency. A screening result was considered to be positive for trisomy 21 if the calculated risk was at least 1 in 270 pregnancies and positive for trisomy 18 if the risk was at least 1 in 150. RESULTS: Screening was completed in 8514 patients with singleton pregnancies. This approach to screening identified 85.2 percent of the 61 cases of Down's syndrome (95 percent confidence interval, 73.8 to 93.0), with a false positive rate of 9.4 percent (95 percent confidence interval, 8.8 to 10.1). At a false positive rate of 5 percent, the detection rate was 78.7 percent (95 percent confidence interval, 66.3 to 88.1). Screening identified 90.9 percent of the 11 cases of trisomy 18 (95 percent confidence interval, 58.7 to 99.8), with a 2 percent false positive rate. Among women 35 years of age or older, screening identified 89.8 percent of fetuses with trisomy 21, with a false positive rate of 15.2 percent, and 100 percent of fetuses with trisomy 18. CONCLUSIONS: First-trimester screening for trisomies 21 and 18 on the basis of maternal age, maternal levels of free beta human chorionic gonadotropin and pregnancy-associated plasma protein A, and measurement of fetal nuchal translucency has good sensitivity at an acceptable false positive rate. PMID- 14534334 TI - A longitudinal, population-based, cohort study of childhood asthma followed to adulthood. AB - BACKGROUND: The outcome of childhood asthma in adults has been described in high risk cohorts, but few population-based studies have reported the risk factors for persistence and relapse. METHODS: We assessed children born from April 1972 through March 1973 in Dunedin, New Zealand, repeatedly from 9 to 26 years of age with questionnaires, pulmonary-function tests, bronchial-challenge testing, and allergy testing. RESULTS: By the age of 26 years, 51.4 percent of 613 study members with complete respiratory data had reported wheezing at more than one assessment. Eighty-nine study members (14.5 percent) had wheezing that persisted from childhood to 26 years of age, whereas 168 (27.4 percent) had remission, but 76 (12.4 percent) subsequently relapsed by the age of 26. Sensitization to house dust mites predicted the persistence of wheezing (odds ratio, 2.41; P=0.001) and relapse (odds ratio, 2.18; P=0.01), as did airway hyperresponsiveness (odds ratio for persistence, 3.00; P<0.001; odds ratio for relapse, 3.03; P<0.001). Female sex predicted the persistence of wheezing (odds ratio, 1.71; P=0.03), as did smoking at the age of 21 years (odds ratio, 1.84; P=0.01). The earlier the age at onset, the greater the risk of relapse (odds ratio, 0.89 per year of increase in the age at onset; P<0.001). Pulmonary function was consistently lower in those with persistent wheezing than in those without persistent wheezing. CONCLUSIONS: In an unselected birth cohort, more than one in four children had wheezing that persisted from childhood to adulthood or that relapsed after remission. The factors predicting persistence or relapse were sensitization to house dust mites, airway hyperresponsiveness, female sex, smoking, and early age at onset. These findings, together with persistently low lung function, suggest that outcomes in adult asthma may be determined primarily in early childhood. PMID- 14534336 TI - Gentamicin-induced correction of CFTR function in patients with cystic fibrosis and CFTR stop mutations. AB - BACKGROUND: Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene containing a premature termination signal cause a deficiency or absence of functional chloride-channel activity. Aminoglycoside antibiotics can suppress premature termination codons, thus permitting translation to continue to the normal end of the transcript. We assessed whether topical administration of gentamicin to the nasal epithelium of patients with cystic fibrosis could result in the expression of functional CFTR channels. METHODS: In a double-blind, placebo-controlled, crossover trial, patients with stop mutations in CFTR or patients homozygous for the DeltaF508 mutation received two drops containing gentamicin (0.3 percent, or 3 mg per milliliter) or placebo in each nostril three times daily for two consecutive periods of 14 days. Nasal potential difference was measured at base line and after each treatment period. Nasal epithelial cells were obtained before and after gentamicin treatment from patients carrying stop mutations, and the C-terminal of surface CFTR was stained. RESULTS: Gentamicin treatment caused a significant reduction in basal potential difference in the 19 patients carrying stop mutations (from -45+/-8 to -34+/-11 mV, P=0.005) and a significant response to chloride-free isoproterenol solution (from 0+/-3.6 to 5+/-2.7 mV, P<0.001). This effect of gentamicin on nasal potential difference occurred both in patients who were homozygous for stop mutations and in those who were heterozygous, but not in patients who were homozygous for DeltaF508. After gentamicin treatment, a significant increase in peripheral and surface staining for CFTR was observed in the nasal epithelial cells of patients carrying stop mutations. CONCLUSIONS: In patients with cystic fibrosis who have premature stop codons, gentamicin can cause translational "read through," resulting in the expression of full-length CFTR protein at the apical cell membrane, and thus can correct the typical electrophysiological abnormalities caused by CFTR dysfunction. PMID- 14534335 TI - Frequency of major molecular responses to imatinib or interferon alfa plus cytarabine in newly diagnosed chronic myeloid leukemia. AB - BACKGROUND: In a randomized trial, 1106 patients with chronic myeloid leukemia (CML) in chronic phase were assigned to imatinib or interferon alfa plus cytarabine as initial therapy. We measured levels of BCR-ABL transcripts in the blood of all patients in this trial who had a complete cytogenetic remission. METHODS: Levels of BCR-ABL transcripts were measured by a quantitative real-time polymerase-chain-reaction assay. Results were expressed relative to the median level of BCR-ABL transcripts in the blood of 30 patients with untreated CML in chronic phase. RESULTS: In patients who had a complete cytogenetic remission, levels of BCR-ABL transcripts after 12 months of treatment had fallen by at least 3 log in 57 percent of those in the imatinib group and 24 percent of those in the group given interferon plus cytarabine (P=0.003). On the basis of the rates of complete cytogenetic remission of 68 percent in the imatinib group and 7 percent in the group given interferon plus cytarabine at 12 months, an estimated 39 percent of all patients treated with imatinib but only 2 percent of all those given interferon plus cytarabine had a reduction in BCR-ABL transcript levels of at least 3 log (P<0.001). For patients who had a complete cytogenetic remission and a reduction in transcript levels of at least 3 log at 12 months, the probability of remaining progression-free was 100 percent at 24 months, as compared with 95 percent for such patients with a reduction of less than 3 log and 85 percent for patients who were not in complete cytogenetic remission at 12 months (P<0.001). CONCLUSIONS: The proportion of patients with CML who had a reduction in BCR-ABL transcript levels of at least 3 log by 12 months of therapy was far greater with imatinib treatment than with treatment with interferon plus cytarabine. Patients in the imatinib group with this degree of molecular response had a negligible risk of disease progression during the subsequent 12 months. PMID- 14534337 TI - Images in clinical medicine. Spindle-cell thymoma. PMID- 14534338 TI - Clinical practice. Combination estrogen-progestin oral contraceptives. PMID- 14534339 TI - Chronic myeloid leukemia--advances in biology and new approaches to treatment. PMID- 14534340 TI - Clinical problem-solving. Mind the gap. PMID- 14534341 TI - Screening for Down's syndrome--too many choices? PMID- 14534342 TI - Toward asthma prevention--does all that really matters happen before we learn to read? PMID- 14534343 TI - High-dose chemotherapy for breast cancer. PMID- 14534344 TI - Nitrous oxide and 5,10-methylenetetrahydrofolate reductase. PMID- 14534345 TI - The vulture and stem cells. PMID- 14534346 TI - Stem cells. PMID- 14534347 TI - Case 21-2003: permission for postmortem examination. PMID- 14534348 TI - Case 11-2003: ulcerative colitis and primary sclerosing cholangitis in a 14-year old boy. PMID- 14534349 TI - Residual disease in chronic myeloid leukemia after induction of molecular remission. PMID- 14534350 TI - Images in clinical medicine. Epidural lipomatosis causing spinal cord compression. PMID- 14534351 TI - Identification of metal-binding proteins in human hepatoma lines by immobilized metal affinity chromatography and mass spectrometry. AB - The metalloproteome is defined as the set of proteins that have metal-binding capacity by being metalloproteins or having metal-binding sites. A different metalloproteome may exist for each metal. Mass spectrometric characterization of metalloproteomes provides valuable information relating to cellular disposition of metals physiologically and in metal-associated diseases. We examined the Cu and Zn metalloproteomes in three human hepatoma lines: Hep G2 and Mz-Hep-1, which retain many functional characteristics of normal human hepatocytes, and SK-Hep-1, which is poorly differentiated. Additionally we studied a single specimen of normal human liver and Hep G2 cells depleted in vitro of cellular copper. We used matrix-assisted laser desorption ionization and electrospray ionization quadrupole time-of-flight mass spectrometry to analyze peptide sequences of tryptic digests obtained by either in-gel digestion of metal-binding proteins or peptides on an immobilized metal affinity chromatography column loaded with either Cu or Zn. Mainly high abundance proteins were identified. Cu-binding proteins identified included enolase, albumin, transferrin, and alcohol dehydrogenase as well as certain intracellular chaperone proteins. The Cu metalloproteome was not identical to the Zn metalloproteome. Peptide binding experiments demonstrated that Cu coordination prefers the order of residues histidine > methionine > cysteine. Although the Cu metalloproteome was similar from line to line, subtle differences were apparent. Gel profiling showed more extensive variation in expression of annexin II in SK-Hep-1 and Mz-Hep-1 than in Hep G2 and normal liver tissue. Glycerylphosphorylethanolamine was identified as a post-translational modification at residue Glu-301 of elongation factor 1-alpha in Hep G2. Intracellular copper depletion was associated with loss of the glycerylphosphoryl side group. These findings suggest that post-translational modification could be affected by intracellular actions of copper. Comparison of the Cu and Zn metalloproteomes in Hep G2 with a published general proteome of Hep G2 disclosed little overlap (Seow, T. K., et al. (2001) Proteomics 1, 1249-1263). Proteins in the metalloproteomes of human hepatocytes can be identified by these methods. Variations in these metalloproteomes may have important physiological relevance. PMID- 14534352 TI - A proteomic analysis of arginine-methylated protein complexes. AB - Arginine methylation is a post-translational modification that results in the formation of asymmetrical and symmetrical dimethylated arginines (a- and sDMA). This modification is catalyzed by type I and II protein-arginine methyltransferases (PRMT), respectively. The two major enzymes PRMT1 (type I) and PRMT5 (type II) preferentially methylate arginines located in RG-rich clusters. Arginine methylation is a common modification, but the reagents for detecting this modification have been lacking. Thus, fewer than 20 proteins have been identified in the last 40 years as containing dimethylated arginines. We have generated previously four arginine methyl-specific antibodies; ASYM24 and ASYM25 are specific for aDMA, whereas SYM10 and SYM11 recognize sDMA. All of these antibodies were generated by using peptides with aDMA or sDMA in the context of different RG-rich sequences. HeLa cell extracts were used to purify the protein complexes recognized by each of the four antibodies, and the proteins were identified by microcapillary reverse-phase liquid chromatography coupled on line with electrospray ionization tandem mass spectrometry. The analysis of two tandem mass spectra for each methyl-specific antibody resulted in the identification of over 200 new proteins that are putatively arginine-methylated. The major protein complexes that were purified include components required for pre-mRNA splicing, polyadenylation, transcription, signal transduction, and cytoskeleton and DNA repair. These findings provide a basis for the identification of the role of arginine methylation in many cellular processes. PMID- 14534353 TI - Chronic ethanol ingestion facilitates N-methyl-D-aspartate receptor function and expression in rat lateral/basolateral amygdala neurons. AB - Withdrawal anxiety after chronic alcohol is likely to contribute to drug seeking and relapse in alcoholics. The brain regions regulating fear/anxiety behaviors, especially neurotransmitter systems with acute ethanol sensitivity, are potential targets for chronic ethanol-induced adaptations. We have therefore examined N methyl-d-aspartate (NMDA) receptors after chronic ethanol ingestion in rat lateral/basolateral amygdala. Whole cell patch-clamp measurements indicate that chronic ethanol ingestion significantly increased NMDA receptor current density. This enhanced NMDA receptor function was also associated with an increase in ifenprodil inhibition and a decrease in apparent calcium-dependent current inactivation. These findings suggest that NR2B-containing receptors may be specifically enhanced and suggest that processes dependent upon calcium influx through amygdala NMDA receptors may potentially be enhanced by chronic ethanol ingestion. We measured subunit mRNA expression to investigate possible molecular mechanisms that control functional receptor adaptations to chronic ethanol. Quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) demonstrated that NR1 subunit mRNA expression, but not NR2 or NR3 expression, was enhanced in samples from chronic ethanol-exposed animals. Single-cell RT-PCR was then used to confirm that NR2 mRNA expression was unaltered by chronic ethanol. Most GAD-, presumed projection neurons expressed both NR2A and NR2B mRNAs, and this profile did not change during chronic ethanol exposure. Our results suggest that both transcriptional and nontranscriptional adaptations to chronic ethanol ultimately contribute to alterations in NMDA receptor function. Because amygdala NMDA receptors play a significant role in many learned fear behaviors, chronic ethanol-induced adaptations in these receptors may influence the expression of withdrawal anxiety. PMID- 14534355 TI - A region-specific increase in Galphaq and Galpha11 proteins in brains of rats during cocaine withdrawal. AB - Serotonin 2A (5-HT2A) receptor-mediated increases in plasma hormone levels become supersensitive after 42 h of withdrawal from cocaine treatment. The present study investigated which components of the 5-HT2A receptor signaling system are associated with this supersensitivity. Rats were injected daily for 14 days with either saline or cocaine (15 mg/kg i.p.) twice a day or were injected using a "binge" protocol (three injections per day, 1 h apart). Rats were sacrificed 2 or 7 days after the last cocaine injection, and the levels of membrane and cytosol associated 5-HT2A receptors, Galphaq, Galpha11, regulators of G protein signaling (RGS)4, and RGS7 proteins were assayed in the hypothalamic paraventricular nucleus, amygdala, and frontal cortex using Western blot analysis. Two days of withdrawal from cocaine, administered twice a day or using a binge protocol, produced an increase in membrane-associated Galphaq and Galpha11 proteins in the paraventricular nucleus and the amygdala (but not in the frontal cortex). This effect was reversible after 7 days of withdrawal. The protein levels of the 5 HT2A receptor, Galphaz protein, and RGS4 or RGS7 proteins were not altered by cocaine withdrawal in any of the above-mentioned brain regions. These findings suggest that the supersensitivity of the 5-HT2A receptors, during withdrawal from chronic cocaine, is associated with an increase in membrane-associated Galphaq and Galpha11 proteins and not with changes in the expression of 5-HT2A receptors. PMID- 14534354 TI - Simultaneous modeling of abciximab plasma concentrations and ex vivo pharmacodynamics in patients undergoing coronary angioplasty. AB - An integrated structural pharmacokinetic/pharmacodynamic (PK/PD) model was developed for the glycoprotein IIb/IIIa antagonist abciximab administered to patients undergoing percutaneous transluminal coronary angioplasty. PK/PD data, in the form of plasma abciximab concentrations and ex vivo platelet aggregation in the presence of 20 microM adenosine diphosphate, were obtained from two previously conducted clinical studies. Study 1 consisted of patients who were given abciximab as a single intravenous injection of 0.25 mg/kg (n = 32). Patients in study 2 received an identical bolus dose, followed by a 36-h infusion at 0.125 microg/kg/min (n = 15). The PK component of the final model included drug-receptor binding, nonspecific distribution, and linear systemic clearance, whereas the PD module assumed that ex vivo dynamics were controlled by free plasma drug concentration. Mean PK/PD data from both studies were fitted simultaneously using nonlinear regression. PK profiles from both studies show rapidly decreasing plasma abciximab concentrations at early time points, but with extended terminal disposition phases. The maximum effect (Emax = 83.6%) was achieved rapidly and gradually returned to baseline values, although inhibition could be measured long after abciximab concentrations dropped below the detection limit. The final model well described the resulting PK/PD profiles and allowed for parameter estimation with relatively small coefficients of variation. Simulations were conducted to assess predicted receptor-occupancy and effects of selected parameters on PK/PD profiles. Models such as the one developed in this study demonstrate how drug binding to pharmacological targets may influence the PK of certain drugs and also provide a suitable paradigm for defining the PK/PD relationships of similar compounds. PMID- 14534356 TI - Pumping of drugs by P-glycoprotein: a two-step process? AB - The apparent inhibition constant, Kapp, for the blockade of P-glycoprotein (P-gp) by four drugs, verapamil, cyclosporin A, XR9576 (tariquidar), and vinblastine, was measured by studying their ability to inhibit daunorubicin and calcein-AM efflux from four strains of Ehrlich cells with different levels of drug resistance and P-gp content. For daunorubicin as a transport substrate, Kapp was independent of [P-gp] for verapamil but increased strictly linearly with [P-gp] for vinblastine, cyclosporin A, and XR9576. A theoretical analysis of the kinetics of drug pumping and its reversal shows that Kapp for inhibition should increase linearly with the amount of pumps present in the membrane for a reverser that inhibits pumping from the cytoplasmic face. In contrast, if the reverser acts by blocking transport from the outer face, i.e., preemptively, Kapp should be independent of the number of pumps present. The experimental data suggest that verapamil blocks pumping at the extracellular face of the membrane, whereas the other three blockers act on pumping from the cytoplasmic phase. The maximum degree of inhibition was the same for all four blockers; thus, they do not act in parallel but rather, in serial, i.e., a drug that is pumped from the cytoplasmic phase has to pass the preemptive route upon leaving the cell. Our results are consistent with the Sauna-Ambudkar two-step model for pumping by P-gp. We suggest that the vinblastine/cyclosporin A/XR9576-binding site accepts daunorubicin at the cytoplasmic face and transfers it to the verapamil-binding site, from where daunorubicin is emptied at the extracellular surface. PMID- 14534357 TI - A novel potent radical scavenger, 8-(4-fluorophenyl)-2-((2E)-3-phenyl-2 propenoyl)-1,2,3,4-tetrahydropyrazolo[5,1-c] [1,2,4]triazine (FR210575), prevents neuronal cell death in cultured primary neurons and attenuates brain injury after focal ischemia in rats. AB - Reactive oxygen species (ROS) play a vital role in brain damage after cerebral ischemia-reperfusion injury, and ROS scavengers have been shown to exert neuroprotective effects against ischemic brain injury. We have recently identified 8-(4-fluorophenyl)-2-((2E)-3-phenyl-2-propenoyl)-1,2,3,4 tetrahydropyrazolo[5,1-c][1,2,4]triazine (FR210575) as a novel, powerful free radical scavenger. In the present study, the neuroprotective efficacy of FR210575 was evaluated in two neuronal death models in vitro as well as rat focal cerebral ischemia models in vivo. In the first model, primary cortical cultures were exposed to a high oxygen atmosphere (50% O2) for 48 h to induce cell death with apoptotic features. Treatment with FR210575 (10-7-10-5 M) significantly inhibited neuronal death. The second model used a growth-factor withdrawal paradigm. Withdrawal of TIP (transferrin, insulin, putrescine and progesterone) supplemented medium induced apoptotic cell death after 2 days, but treatment with FR210575 exhibited dramatic protection against neuronal death. In two models of cerebral ischemia [photothrombotic occlusion of middle cerebral artery (MCA) for transient model and by permanent MCA occlusion for permanent model], rats received 3-h intravenous infusion (1-10 mg/kg/3 h) of FR210575, with brain damage determined 24 h later. FR210575 (3.2 mg/kg/3 h) significantly reduced the volume of focal damage in the cortex by 36% in the transient model and also reduced the size of ischemic brain damage in the permanent model. These findings indicate that the powerful radical scavenger FR210575 has potent neuroprotective activity and that FR210575 could be an attractive candidate for the treatment of stroke or other neurodegenerative disorders. PMID- 14534358 TI - Evidence for the involvement of a pulmonary first-pass effect via carboxylesterase in the disposition of a propranolol ester derivative after intravenous administration. AB - The disposition kinetics of O-butyryl propranolol (butyryl-PL), a model compound containing an ester moiety, after intravenous administration was compared with that of PL in rats and beagle dogs. Rats showed only 30% conversion of butyryl-PL to PL up to 2 h after dosing, whereas dogs showed nearly complete conversion within 10 min after administration. The CL(total) of butyryl-PL in rats was 5.8 l/h/kg and that in dogs was 65.6 +/- 18.6 l/h/kg, both of which were greater than hepatic blood flow. The in vivo conversion from butyryl-PL to PL in the rat could be explained on the basis of the hydrolysis characteristics in the liver and blood. The in vitro hydrolysis data and the in vivo data after intra-arterial administration clearly demonstrated that the extremely high CL(total) of butyryl PL in dogs was dependent on first-pass hydrolysis in the lung in addition to hydrolysis at a blood flow-limited rate in the liver and kidney. The availability of butyryl-PL after passage through the lung was 50%. Furthermore, the isoform of carboxylesterase involved in the pulmonary hydrolysis of butyryl-PL in the dog was identified as D1, a CES-1 group enzyme. However, butyryl-PL was not recognized as a substrate by CES-1 family carboxylesterases, which are present at high levels in the rat lung (RH-1) and kidney (RL-1). These findings indicate that extrahepatic metabolism, especially in the lung, is important in the disposition of drugs containing an ester moiety after intravenous administration and that the substrate specificity of carboxylesterase isozyme distinguishes from others. PMID- 14534359 TI - Identification of critical residues in the amino terminal domain of the human NR2B subunit involved in the RO 25-6981 binding pocket. AB - N-Methyl-d-aspartate (NMDA) receptors play key roles in both physiological processes, particularly synaptic plasticity, and in neuropathological states such as epilepsy and acute neurodegeneration. R-(R*,S*)-alpha-(4-Hydroxyphenyl)-beta methyl-4-(phenyl-methyl)-1-piperidine propanol (RO 25-6981), is a high-affinity and selective blocker of NMDA receptors containing the NR2B subunit. Using site directed mutagenesis, [3H]RO 25-6981 binding, Xenopus oocyte voltage-clamp recordings, and molecular modeling, we have identified several critical residues involved in the RO 25-6981 binding site within the N-terminal LIVBP-like domain of the human NR2B subunit. Two mutations, NR2B(D101A) and NR2B(F176A), resulted in a complete loss of [3H]RO 25-6981 binding and also abolished the high-affinity RO 25-6981-mediated inhibition of NMDA-induced currents. The mutation NR2B(T233A) led to a marked reduction in binding affinity by 13-fold. Mutations F182A, D104A, or K234A had a more moderate influence on the binding affinity (KD values increased by 8-, 7-, and 6-fold, respectively). In a three-dimensional model of the NR2B LIVBP-like domain based on the X-ray crystal structure of the amino terminal domain of the mGlu1 receptor, the critical residues are located in the central cleft where interaction with RO 25-6981 may stabilize the closed structure of the domain. Our results suggest that the three amino acids Asp-101, Phe-176, and Thr-233 are important molecular determinants for the high-affinity binding of RO 25-6981 to the LIVBP-like domain of human NR2B. A possible binding mode for RO 25-6981 is proposed. PMID- 14534360 TI - Fructose-fed rats are protected against ischemia/reperfusion injury. AB - This study examines the relationship between insulin resistance (IR) induced by fructose feeding (FF) and susceptibility to myocardial ischemia/reperfusion injury (MI/R). Six-week-old male Sprague-Dawley rats were randomized into control (CON; n = 59) or FF (n = 58) groups. After 4 weeks, rats were further randomized into one of the following groups: placebo, ischemic preconditioning (IPC), 5 hydroxydecanoic acid (5-HD) (10 mg/kg), or 5-HD + IPC. Moreover, to determine the role of fructose, a second model of IR (Zucker obese) and rats fed fructose diet for 3 days (FF-3) were also subjected to MI/R. In all experiments, rats were subjected to 30 min of myocardial ischemia and 4 h of reperfusion. In rats randomized to placebo, infarct size was significantly reduced by FF (24 +/- 5%) compared with CON (54 +/- 1%, p < 0.05). Pretreatment with 5-HD did not alter the infarct size in CON (45 +/- 5%) but inhibited the protection afforded by FF (53 +/- 7%). IPC reduced the infarct size to an equivalent level in both groups, whereas 5-HD administration prior to IPC blunted the IPC effect. In Zucker obese rats, infarct size was significantly larger (57 +/- 4%) compared with lean controls (37 +/- 4%, p < 0.05). In FF-3 rats, infarct size was also decreased (20 +/- 2%, p < 0.01) compared with CON. This study suggests that fructose feeding affords protection against MI/R that is related to or mimics preconditioning. This protection is not consistent with other models of IR and is likely related to the fructose diet itself. PMID- 14534361 TI - Amiodarone inhibits sarcolemmal but not mitochondrial KATP channels in Guinea pig ventricular cells. AB - ATP-sensitive K(+) (KATP) channels are present on the sarcolemma (sarcKATP channels) and mitochondria (mitoKATP channels) of cardiac myocytes. Amiodarone, a class III antiarrhythmic drug, reduces sudden cardiac death in patients with organic heart disease. The objective of the present study was to investigate the effects of amiodarone on sarcKATP and mitoKATP channels. Single sarcKATP channel current and flavoprotein fluorescence were measured in guinea pig ventricular myocytes to assay sarcKATP and mitoKATP channel activity, respectively. Amiodarone inhibited the sarcKATP channel currents in a concentration-dependent manner without affecting its unitary amplitude. The IC50 values were 0.35 microM in the inside-out patch exposed to an ATP-free solution and 2.8 microM in the cell-attached patch under metabolic inhibition, respectively. Amiodarone (10 microM) alone did not oxidize the flavoprotein. In addition, the oxidative effect of the mitoKATP channel opener diazoxide (100 microM) was unaffected by amiodarone. Exposure to ouabain (1 mM) for 30 min produced mitochondrial Ca(2+) overload, and the intensity of rhod-2 fluorescence increased to 246 +/- 16% of baseline (n = 9). Amiodarone did not alter the ouabain-induced mitochondrial Ca(2+) overload (236 +/- 10% of baseline, n = 7). Treatment with diazoxide significantly reduced the ouabain-induced mitochondrial Ca(2+) overload (158 +/- 15% of baseline, n = 8, p < 0.05 versus ouabain); this effect was not abolished by amiodarone (154 +/- 10% of baseline, n = 8, p < 0.05 versus ouabain). These results suggest that amiodarone inhibits sarcKATP but not mitoKATP channels in cardiac myocytes. Such an action of amiodarone may effectively prevent ischemic arrhythmias without causing ischemic damage. PMID- 14534362 TI - Site of action of the general anesthetic propofol in muscarinic M1 receptor mediated signal transduction. AB - Although a potential target site of general anesthetics is primarily the GABA A receptor, a chloride ion channel, a previous study suggested that the intravenous general anesthetic propofol attenuates the M1 muscarinic acetylcholine receptor (M1 receptor)-mediated signal transduction. In the present study, we examined the target site of propofol in M1 receptor-mediated signal transduction. Two electrode voltage-clamp method was used in Xenopus oocytes expressing both M1 receptors and associated G protein alpha subunits (Gqalpha). Propofol inhibited M1 receptor-mediated signal transduction in a dose-dependent manner (IC50 = 50 nM). Injection of guanosine 5'-3-O-(thio)triphosphate (GTPgammaS) into oocytes overexpressing Gqalpha was used to investigate direct effects of propofol on G protein coupled with the M1 receptor. Propofol did not affect activation of Gqalpha-mediated signal transduction with the intracellular injection of GTPgammaS. We also studied effects of propofol on l-[N-methyl-3H]scopolamine methyl chloride ([3H]NMS) binding and M1 receptor-mediated signal transduction in mammalian cells expressing M1 receptor. Propofol inhibited the M1 receptor mediated signal transduction but did not inhibit binding of [3H]NMS. Effects of propofol on Gs- and Gi/o-coupled signal transduction were investigated, using oocytes expressing the beta2 adrenoceptor (beta2 receptor)/cystic fibrosis transmembrane conductance regulator or oocytes expressing the M2 muscarinic acetylcholine receptor (M2 receptor)/Kir3.1 (a member of G protein-gated inwardly rectifying K(+) channels). Neither beta2 receptor-mediated nor M2 receptor mediated signal transduction was inhibited by a relatively high concentration of propofol (50 microM). These results indicate that propofol inhibits M1 receptor mediated signal transduction by selectively disrupting interaction between the receptor and associated G protein. PMID- 14534363 TI - Agonist-dependent internalization of the human melanocortin-4 receptors in human embryonic kidney 293 cells. AB - A chimeric protein comprised of melanocortin-4 receptor (MC4R) and the green fluorescent protein (GFP) was created for studying receptor/ligand localization and trafficking. The ligand binding affinities and second messenger stimulation induced by MC4R-GFP closely resembled those of the wild-type receptor, suggesting functional integrity of the chimeric protein. As observed with a confocal microscope, in human embryonic kidney (HEK)-293 cells MC4R/GFP was distributed evenly along the cell membrane. Addition of [Nle4-d-Phe7]-alpha-melanocyte stimulating hormone (NDP-MSH), a peptide MC4R agonist, induced receptor translocation into intracellular compartments in a time- and concentration dependent manner. [Ac-Nle-c[Asp-His-d-Nal(2')-Arg-Trp-Lys]-NH2] (SHU9119), a potent MC4R antagonist, completely inhibited NDP-MSH-mediated internalization. MC4R-GFP internalization was unaffected by a protein kinase A inhibitor N-[2-(p bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide (H89), but was impaired by pretreatment with inhibitors of endocytosis through clathrin-coated pits, hypertonic sucrose, or concanavalin A. Time-dependent colocalization of MC4R-GFP with rhodamine-transferrin, an early endosome marker, and with LysoTraker, a lysosome marker, was observed after short-term (45 min) and prolonged (20 h) agonist exposure, respectively. Rhodamine-[AcNle-c[Asp-His-d-Phe-Arg-Trp-Lys] NH2] (MTII), a fluorescent derivative of an MC4R agonist, was found to cointernalize with MC4R-GFP into intracellular vesicles. No significant receptor recycling or segregation from the ligand was observed 60 min after removal of the agonist. In contrast, an antagonist rhodamine-Ac-Cys-Glu-His-(d-Nal)-Arg-Trp-Gly Cys-Pro-Pro-Lys-Asp-NH2 (HS014) bound to and colocalized with MC4R-GFP on the cell surface and did not stimulate receptor internalization. In sum, these results suggest that MC4R is subject to agonist-dependent endocytosis via clathrin-coated pits. Prolonged agonist exposure directs MC4R into lysosomes, possibly for degradation. Receptor and ligand recycling is not efficient for MC4R in HEK-293 cells. PMID- 14534364 TI - Renal extraction of angiotensin II. AB - Angiotensin II regulates many aspects of renal function and thereby influences long-term blood pressure. The effects of angiotensin II on the kidney have been exhaustively studied; however, the converse (i.e., effects of the kidney on angiotensin II) has received little attention. Accordingly, the focus of this study was to determine whether renal degradation of angiotensin II is regulated by chronic levels of angiotensin II or long-term levels of blood pressure. Twenty hypertensive rats and 22 normotensive rats were treated for 1 week with either vehicle, angiotensin II (50 ng/kg/min, subcutaneously) or captopril (100 mg/kg/day, orally). Right kidney vascular resistance was measured during infusions of angiotensin II into the left renal artery or vena cava at the level of left renal vein. Dose-response data were curve-fitted, and the extraction of angiotensin II by the left kidney was calculated by comparing the doses of angiotensin II required to elicit equal increases in right renal vascular resistance during intravenous versus left intrarenal artery infusions. Renal extraction of angiotensin II was high (mean, 81%) and demonstrated little animal to-animal variation (coefficient of variation, 23%; standard deviation, 19%). Renal extraction of angiotensin II was independent of hypertension (P = 0.257) or previous chronic exposure to angiotensin II or captopril (P = 0.270), and there was no interaction between hypertension and chronic exposure to angiotensin II or captopril (P = 0.950). We conclude that renal degradation of angiotensin II is constitutively high, is unaffected by chronic levels of arterial blood pressure, and is independent of long-term changes in levels of angiotensin II. PMID- 14534365 TI - Steroid hormone interactions with target cells: cross talk between membrane and nuclear pathways. AB - The biological effects of steroid hormones are mediated by receptors associated with the plasma membrane as well as located inside of target cells. This perspective focuses on recent advances in our understanding of the integration that occurs between membrane-associated rapid signaling events and various changes in gene transcription that modulate the function and phenotype of steroid responsive cells. Three frequently studied members of the steroid hormone receptor superfamily, the estrogen receptors, the thyroid hormone receptors, and the vitamin D receptors, are included to illustrate the emerging concepts. Each of these hormones has been conclusively shown to function at multiple subcellular sites leading to a continuum of signals intimately linked by intracellular cross talk. Understanding the molecular mechanisms by which these steroid hormones and their receptors transduce cellular signals will allow us to create new pharmacologic therapies aimed at treatment of a variety of human diseases affecting the cardiovascular system, the reproductive system, the skeletal system, the nervous system, the mammary gland, and many others. PMID- 14534366 TI - Comparison of [Dmt1]DALDA and DAMGO in binding and G protein activation at mu, delta, and kappa opioid receptors. AB - [Dmt1]DALDA (H-Dmt-d-Arg-Phe-Lys-NH2; Dmt = 2',6'-dimethyltyrosine) binds with high affinity and selectivity to the mu opioid receptor and is a surprisingly potent and long-acting analgesic, especially after intrathecal administration. In an attempt to better understand the unique pharmacological profile of [Dmt1]DALDA, we have prepared [3H][Dmt1]DALDA and compared its binding properties with that of [3H]DAMGO ([d-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin). Kinetic studies revealed rapid association of [3H][Dmt1]DALDA when incubated with mouse brain membranes (K+1 = 0.155 nM(-1) min(-1)). Dissociation of [3H][Dmt1]DALDA was also rapid (K(-1) = 0.032 min(-1)) and indicated binding to a single site. [3H][Dmt1]DALDA binds with very high affinity to human mu opioid receptor (hMOR) (Kd = 0.199 nM), and Kd and Bmax were reduced by sodium but not Gpp(NH)p [guanosine 5'-(beta,gamma-imido)triphosphate]. Similar Kd values were obtained in brain and spinal cord tissues and SH-SY5Y cells. The hMOR:hDOR (human delta opioid receptor) selectivity of [Dmt1]DALDA ( approximately 10,000) is 8-fold higher than DAMGO. However, [Dmt1]DALDA is less selective than DAMGO against hKOR (human kappa opioid receptor) (26-versus 180-fold). The Ki values for a number of opioid ligands were generally higher when determined by competitive displacement binding against [3H][Dmt1]DALDA compared with [3H]DAMGO, with the exception of Dmt1-substituted peptide analogs. All Dmt1 analogs showed much higher affinity for the mu receptor than corresponding Tyr1 analogs. [35S]GTPgammaS (guanosine 5' O -(3-[35S]thio)triphosphate) binding showed that [Dmt1]DALDA and DAMGO are full agonists at hMOR and hDOR but are only partial agonists at hKOR. The very high affinity and selectivity of [3H][Dmt1]DALDA for the mu receptor, together with its very low nonspecific binding (10-15%) and metabolic stability, make [3H][Dmt1]DALDA an ideal radioligand for labeling mu receptors. PMID- 14534367 TI - DPI-3290 [(+)-3-((alpha-R)-alpha-((2S,5R)-4-Allyl-2,5-dimethyl-1-piperazinyl)-3 hydroxybenzyl)-N-(3-fluorophenyl)-N-methylbenzamide]. II. A mixed opioid agonist with potent antinociceptive activity and limited effects on respiratory function. AB - Allyl-2,5-dimethyl-1-piperazines have been of interest as analgesic agents for the management of moderate-to-severe pain. In this study, we compared the antinociceptive properties and respiratory depressant activity of one such agent, (+)-3-((alpha-R)-alpha-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3 hydroxybenzyl)-N-(3-fluorophenyl)-N-methylbenzamide (DPI-3290), with those of established narcotic analgesics, morphine and fentanyl. Intravenous administration of DPI-3290 in conscious laboratory rats increased antinociception in a dose-dependent manner with a corresponding ED(50) value of 0.05 +/- 0.0072 mg/kg. Simultaneous measurement of arterial blood gas in animals treated with DPI 3290 demonstrated dose-dependent increases in pCO2 with an ED(50) value of 0.91 +/- 0.22 mg/kg. In comparison, morphine and fentanyl increased antinociception in rats with ED(50) values of 2.01 +/- 0.0005 and 0.0034 +/- 0.00024 mg/kg, respectively, and the ED(50) value for morphine-induced changes in pCO2 was 4.23 +/- 0.72 mg/kg, whereas the ED(50) value for fentanyl-induced changes in pCO2 was 0.0127 +/- 0.0035 mg/kg. A separate series of experiments were designed to examine the effects of DPI-3290 on mu-opioid receptor induced antinociception and hypercapnia. Intravenous bolus doses of DPI-3290 that ranged from 0.2 to 1.0 mg/kg had no effect on antinociception mediated by alfentanil (2 microg/kg/min i.v.) but reduced hypercapnia by approximately 50%. Results from these studies demonstrate the equivalent antinociceptive efficacy of DPI-3290, morphine, and fentanyl but dramatic differences in the hypercapnia that antinociceptive doses of these drugs produce. When measured simultaneously, DPI-3290 had an 18.2-fold difference in the ratio comparing the ED(50) value for antinociception with the ED(50) value for changes in pCO2; this ratio was 2.1 for morphine and 3.7 for fentanyl. Furthermore, DPI-3290 reduced the alfentanil-mediated hypercapnia without any effect on antinociception. Together, the balanced opioid agonist activity of DPI-3290 may provide a means of powerful analgesia while mitigating the mu-opioid receptor-mediated hypercapnia. PMID- 14534368 TI - DPI-3290 [(+)-3-((alpha-R)-alpha-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3 hydroxybenzyl)-N-(3-fluorophenyl)-N-methylbenzamide]. I. A mixed opioid agonist with potent antinociceptive activity. AB - Compound (+)-3-((alpha-R)-alpha-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3 hydroxybenzyl)-N-(3-fluorophenyl)-N-methylbenzamide (DPI-3290), is one of a series of novel centrally acting agents with potent antinociceptive activity that binds specifically and with high affinity to opioid receptors. In saturation equilibrium binding studies performed at 25 degrees C using membranes from rat brain or guinea pig cerebellum, the Ki values measured for DPI-3290 at delta-, mu , and kappa-opioid receptors were 0.18 +/- 0.02, 0.46 +/- 0.05, and 0.62 +/- 0.09 nM, respectively. In vas deferens isolated from laboratory mice, DPI-3290 decreased electrically induced tension development in a concentration-dependent manner with corresponding IC50 values of 1.0 +/- 0.3, 6.2 +/- 2.0, and 25.0 +/- 3.3 nM at delta-, mu-, and kappa-receptors, respectively. The activity of DPI 3290 in isolated vas deferens tissue was approximately 20,000, 175.8, and 1500 times more efficacious than morphine, and 492, 2.5, and 35 times more efficacious than fentanyl at delta-, mu-, and kappa-receptors, respectively. In ileal strips isolated from guinea pigs, DPI-3290 inhibited tension development with a corresponding IC50 value of 3.4 +/- 1.6 nM at mu-opioid receptors and 6.7 +/- 1.6 nM at kappa-opioid receptors. Intravenous administration of 0.05 +/- 0.007 mg/kg DPI-3290 produced a 50% antinociceptive response in rats. The antinociceptive properties of DPI-3290 were blocked by naloxone (0.5 mg/kg s.c.). Compared with morphine, this study demonstrated that DPI-3290 is more potent and elicited a similar magnitude of antinociceptive activity in the rat, actions mediated by its mixed opioid receptor agonist activity. The marked antinociceptive activity of DPI-3290 will likely provide a means for relieving severe pain in patients that require analgesic treatment. PMID- 14534369 TI - Tumor necrosis factor-alpha and troglitazone regulate plasminogen activator inhibitor type 1 production through extracellular signal-regulated kinase- and nuclear factor-kappaB-dependent pathways in cultured human umbilical vein endothelial cells. AB - Plasminogen activator inhibitor type 1 (PAI-1) plays a role in the development of atherosclerosis in diabetic patients. PAI-1 is produced by endothelial cells stimulated with various inflammatory cytokines, such as tumor necrosis factor (TNF)-alpha, which induces insulin resistance. In diabetic patients, troglitazone, a thiazolidinedione, can lower the concentration of PAI-1. We investigated the TNF-alpha-induced signaling pathway that leads to PAI-1 synthesis and the target step of troglitazone in this pathway. TNF-alpha induced PAI-1 mRNA expression and protein production in human umbilical vein endothelial cells (HUVECs). A specific inhibitor for p38 mitogen-activated protein kinase, 4 (4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)1H-imidazole (SB 203580), and a protein kinase C inhibitor, calphostin C, had no inhibitory effects on TNF-alpha-induced PAI-1 secretion. A protein tyrosine kinase inhibitor, genistein, completely inhibited TNF-alpha-induced PAI-1 secretion, whereas an inhibitor of extracellular signal-regulated kinase (ERK) kinase, 2' amino-3'-methoxyflavone (PD98059), and a nuclear factor-kappaB (NF-kappaB) inhibitor, emodin, partly inhibited TNF-alpha-induced PAI-1 secretion. Together, PD98059 and emodin completely inhibited TNF-alpha-induced PAI-1 secretion, suggesting that both NF-kappaB-dependent and NF-kappaB-independent pathways are involved in TNF-alpha-induced signal pathway to PAI-1 production and that the latter pathway is mediated by activation of ERK. Furthermore, we have shown that troglitazone inhibited both TNF-alpha-induced PAI-1 protein secretion and mRNA in HUVECs. Genistein, but neither PD98059 nor emodin, was additive to the inhibitory effect of troglitazone on TNF-alpha-induced PAI-1 secretion. These results indicate That ERK and NF-kappaB are possible targets of TNF-alpha and troglitazone in the regulation of PAI-1 production. PMID- 14534370 TI - Ginger reduces hyperglycemia-evoked gastric dysrhythmias in healthy humans: possible role of endogenous prostaglandins. AB - Acute hyperglycemia evokes gastric slow wave dysrhythmias via endogenous prostaglandin generation. Ginger exhibits slow wave antiarrhythmic effects in other models, but its actions on hyperglycemia-evoked gastric dysrhythmias are unexplored. We hypothesized that ginger prevents disruption of slow wave rhythm by acute hyperglycemia via inhibition of prostaglandin production but not its actions. Twenty-two healthy humans underwent fasting electrogastrography during hyperglycemic clamping to 250 to 290 mg/dl after double-blind placebo or ginger root (1 g). Responses were compared with the prostaglandin E1 analog misoprostol (400 microg). Dominant frequencies (DF) and the percentage of recording times in the bradygastric [0.5-2 cycles/min (cpm)], normal (2-4 cpm), and tachygastric (4 9 cpm) frequency ranges were analyzed. After placebo, hyperglycemia reduced normal 2 to 4 cpm activity from 94.4 +/- 2.6 to 66.0 +/- 10.4%, increased the DF from 2.96 +/- 0.04 to 4.09 +/- 0.45 cpm, and increased tachygastria from 2.0 +/- 1.4 to 29.3 +/- 10.7% (P < 0.05). Hyperglycemia effects on normal activity (77.3 +/- 8.3%), DF (3.46 +/- 0.37 cpm), and tachygastria (15.6 +/- 8.6%) were significantly reduced by ginger (P < 0.05). Misoprostol evoked decreases in normal activity from 95.4 +/- 2.0 to 81.7 +/- 3.0% and increases in tachygastria from 3.1 +/- 1.6 to 11.2 +/- 2.4% (P < 0.05). However, ginger did not correct these abnormalities versus placebo (P = N.S.). In conclusion, acute hyperglycemia evokes gastric slow wave dysrhythmias that are prevented by ginger root. Conversely, the compound has no effect on dysrhythmias elicited by a prostaglandin E(1) analog, indicating that ginger likely acts to blunt production of prostaglandins rather than inhibiting their action. These findings suggest novel mechanisms for the traditional Chinese herbal remedy ginger. PMID- 14534371 TI - Pregnancy outcomes following the diagnosis of interstitial cystitis. AB - OBJECTIVE: To describe the outcome of pregnancy in patients treated for interstitial cystitis. METHODS: Over a 4-year period, 12 patients with interstitial cystitis who had one course of intravesical dimethyl sulphoxide (DMSO) therapy for interstitial cystitis were studied. Information on the effects of the treatment-to-conception interval, symptom control during pregnancy and immediately post partum, other pregnancy events and pregnancy outcome were collected. RESULTS: The mean age was 32 years (SD = 5.4), and 6 (50%) were nulliparous. Fifty-eight percent of the women presented with combined urinary and pain symptoms, 25% with a pain symptom only and 17% with a urinary symptom only. Eleven women had 1 course (6 doses) of DMSO therapy. Symptom relief was maintained during pregnancy in almost all the women. The mean interval from diagnosis of interstitial cystitis to the first day of the last menstrual cycle preceding all pregnancies beyond 28 weeks was 19.3 months (SD = 14.7). This interval was 6 months or less for 2 pregnancies that were terminated on account of symptoms attributable to interstitial cystitis. Thirteen healthy babies were delivered. CONCLUSION: Pregnancy outcomes appear good following treatment for interstitial cystitis with 1 course of intravesical DMSO. PMID- 14534372 TI - Atypical endometrial hyperplasia: grounds for possible misdiagnosis of endometrial adenocarcinoma. AB - OBJECTIVE: The diagnoses of atypical hyperplasia and well-differentiated adenocarcinoma imply totally different approaches because of clinical and patient oriented ramifications, especially when morphological differences are not entirely conclusive. The purpose of this study was to examine the relationship between the diagnosis of atypical hyperplasia during curettage or endometrial biopsy and the definitive histological findings from hysterectomy material. STUDY DESIGN: 23 patients were found fit for the current study and subsequently their clinical histories were reviewed for relevant clinical data, histopathological profiling and type of therapeutic interventions. RESULTS: Adenocarcinoma was observed in 12 (52.17%) of 23 hysterectomy cases. The hyperplasia was found in 10 (43.47%) cases, although 4 of them lacked atypia and 1 case proved to be hyperplasia-free. CONCLUSION: Hysterectomy was prescribed as the next step in the diagnosis of atypical endometrial hyperplasia. Other wait-and-see approaches could have easily forfeited the chances of providing an adequate treatment for an operable and curable cancer in approximately half of the studied cases. PMID- 14534373 TI - Gastrostomy tube insertion into intestinal-cutaneous tract fistulas is a new technique to improve fistula control. AB - BACKGROUND: The management of gastrointestinal-cutaneous fistulas may be complicated by the difficulty in obtaining adequate control of the fistula tract. This study describes a new method to obtain better fistula control utilizing a semi-rigid stent in the form of a gastrostomy tube. METHODS: Consecutive patients with intestinal-cutaneous fistulas of at least 3 weeks duration and treated by the new technique were analyzed. The technique involved the insertion of a guide wire into the fistula tract from the luminal side using an endoscope, snaring the wire with a Dormia basket inserted into the fistula tract from the cutaneous side and then exteriorized. The gastrostomy tube was then pulled with the guide wire from the lumen along the fistula tract and out through the skin. RESULTS: Five patients had had fistulas for a median duration of 42 (range 26-140) days before insertion of the gastrostomy tube. The gastrostomy tube was replaced with a smaller diameter tube in 4 of the patients (range 1-3 changes). The patients were discharged from the hospital at a median of 14 (range 12-23) days after the tube insertion but with the tube in situ. The median time from the insertion of the tube to its removal was 42 (range 32-108) days. CONCLUSIONS: Gastrostomy tube insertion using minimally invasive techniques may improve fistula control enabling patients to be discharged home sooner than otherwise and improve the rate of healing. PMID- 14534374 TI - Experimental model of obstructive, chronic pancreatitis in pigs. AB - BACKGROUND: We aimed to develop a reproducible, experimental model of obstructive pancreatitis for future analysis of surgical interventions, and characterized this model using functional, histological and biochemical parameters. ANIMALS AND METHODS: In 10 female pigs the pancreatic duct (PD) was ligated. After 4, 6 or 8 weeks the animals were sacrificed. Before and after ligation, glucose tolerance and intraductal pressure were measured, and pancreatic juice was collected after stimulation with cholecystokinin and secretin. Amylase and lipase activities were analyzed in plasma and juice. Pancreatic tissue was collected for histochemical analysis. RESULTS: Within 4 weeks of ligation, the pancreas appeared atrophic. Intraductal pressure had risen significantly. Acinar-to-ductal metaplasia was accompanied by strong proliferation of stellate cells and increased collagen deposition. Islets of Langerhans appeared smaller and more numerous. Blood amylase and lipase levels were normal and glucose tolerance was unaffected. Pancreatic juice volume and amylase and lipase activities were significantly lower. CONCLUSION: Ligation of the PD in pigs resulted in a marked fibrosing obstructive pancreatitis within 4 weeks, similar to human chronic pancreatitis in regard to clinical, functional, histological and biochemical parameters. PMID- 14534375 TI - Splenectomy versus spleen-preserving surgery for splenic echinococcosis. AB - BACKGROUND/AIMS: The spleen is the third most common location of hydatid disease after liver and lung. The aim of this study was to analyse the long-term outcome of surgical treatment of patients with splenic echinococcosis comparing splenectomy with spleen-preserving surgery. METHODS: During a period of 25 years (1976-2001), 19 (5.4%) patients with splenic echinococcosis were treated in our department out of 349 patients with abdominal hydatid disease. In 16 patients the spleen was the only organ involved, while in 3 patients the liver was also affected. RESULTS: Eleven patients had splenectomy and in the other 8 the spleen was preserved: enucleation (n = 4), partial cystectomy and omentoplasty (n = 2) and cystojejunal Roux-en-Y anastomosis (n = 2). One (6%) patient died in the early postoperative period and 5 (29%) patients had postoperative complications. There was no significant difference between the splenectomy and spleen-preserving groups concerning median hospital stay and postoperative complication rate. The median follow-up in 15 patients was 52 (range 6-300) months. Two patients (13%) developed recurrence of the disease requiring re-operation at 2 and 3 years, respectively. Recurrence occurred in 1 (12%) patient in the splenectomy group and in 1 (14%) out of 7 patients in the spleen-preserving group. CONCLUSION: In the present series it was possible to preserve the spleen in 8 (42%) of 19 patients, without significant increase of recurrent echinococcosis. PMID- 14534376 TI - Serum immunosuppressive acidic protein as an interleukin-6 related index of deteriorating condition in gastric cancer patients. AB - BACKGROUND/AIMS: Immunosuppressive acidic protein (IAP) is an acute-phase reactant which has a close correlation with the impairment of the host's immunity. The present study aims to investigate the significance of serum IAP as an index of cytokine-related disease status in gastric cancer patients. METHODS: Serum IAP levels were determined in 76 gastric cancer patients and 20 healthy subjects. In a subgroup of 39 patients, tissue interleukin-6 (IL-6) concentrations and expression of IL-6 protein in tumor tissues were also examined. RESULTS: The mean serum IAP level in the patients was significantly higher than that in the normal controls. The serum IAP level in the patients was associated with clinicopathological features, such as tumor size and serosal invasion. The prognosis of patients with high IAP levels was significantly worse than that of those with low IAP levels. Moreover, the serum IAP level was closely correlated with various parameters reflecting the host's nutritional and immunological conditions. Immunohistochemically, IL-6 was overexpressed in the cytoplasm of tumor cells. The IL-6 concentration and immunoreactivity of IL-6 protein in tumor tissue was significantly correlated with the serum IAP level. CONCLUSIONS: Elevated serum IAP, which may be upregulated by an activated IL-6 network in tumor tissue, may reflect not only tumor progression, but also a deteriorated condition that is associated with malnutrition and immunosuppression in gastric cancer patients. PMID- 14534377 TI - Evaluation of the POSSUM scoring system for comparative audit in pancreatic surgery. AB - BACKGROUND: Wide variations exist in the reported morbidity and mortality rates for major pancreatic resections. The Physiological and Operative Scoring System for enUmeration of Morbidity and mortality (POSSUM) was developed for comparative audit in general surgical patients. It has also been found to be reliable for audit in colorectal, thoracic and vascular surgery with minor modifications. AIMS: To evaluate POSSUM and its modification for mortality, P-POSSUM, in pancreatic surgery. METHODS: Retrospective analysis of 50 patients undergoing partial pancreaticoduodenectomy (PD) (46 tumours, 4 chronic pancreatitis) using the POSSUM and P-POSSUM as predictors of morbidity and mortality. These were then compared with the observed values. RESULTS: The POSSUM-predicted mortality was 26%. The P-POSSUM predicted a mortality risk of 6%. The observed mortality was 4%. Using POSSUM for morbidity, the predicted value was 76%. The observed morbidity was 46%. The risk scores for patients with and without morbidity were similar (66.4 +/- 11.0 vs. 68.8 +/- 12.9, p = 0.49). CONCLUSIONS: While P-POSSUM appeared satisfactory for predicting mortality risk, POSSUM overestimated morbidity and mortality for PD in a specialist centre. Modifications are needed prior to its application for comparative audit in pancreatic surgery. PMID- 14534378 TI - Ivor Lewis (1895-1982) - Welsh pioneer of the right-sided approach to the oesophagus. PMID- 14534379 TI - Incident cerebrovascular disease in rural and urban Alberta. AB - STUDY OBJECTIVE: This study examines the pattern of incidence and health service utilisation of cerebrovascular disease cases in urban and rural areas within a publicly funded health care system. DESIGN: A population-based study covering a large geographic region, using population-wide administrative health data. Age- and sex-standardised incidence and mortality rates were calculated for rural and urban areas. Final status (discharge or death), place of service and place of residence were reported for all cases across several different subsets of cerebrovascular disease. SETTING: The province of Alberta, located in western Canada. PARTICIPANTS: Incident cases of cerebrovascular disease (stroke and transient ischaemic attack) and 4 different definitions of incident stroke were identified from data on emergency department admissions in the 1999/2000 fiscal year. MAIN RESULTS: The rate of cerebrovascular disease per 10,000 was similar between urban (13.24) and rural (13.82) areas. Rural residents frequently reported their incident episode to urban emergency departments. Although the mortality is similar between urban and rural residents, rural dwellers die more frequently in the emergency department setting than urban dwellers, who die more often as in-patients. CONCLUSIONS: Overall mortality is similar between urban and rural residents. A large proportion of rural residents receive diagnoses and treatment for cerebrovascular disease in urban areas. Location of service and location of death differs between rural and urban cases of cerebrovascular disease. PMID- 14534380 TI - Cerebral vasculitis with multiple infarcts caused by lyme disease. PMID- 14534381 TI - Wegener's granulomatosis presenting with intracerebral hemorrhages. PMID- 14534382 TI - Focal thrombi in the common carotid artery. PMID- 14534383 TI - Superficial siderosis of the brain as a late complication of subarachnoid haemorrhage. PMID- 14534384 TI - Economy class stroke syndrome. PMID- 14534385 TI - Components of difference in HIV seropositivity rate among injection drug users between low- and high-HIV-prevalence regions. AB - Comparative studies on regional HIV seroprevalence or seropositivity rate among injection drug users (IDUs) have focused primarily on assessing the risk factors for HIV infection. This study used a nonparametric analytic approach, known as standardization and decomposition, to to compare HIV seropositivity rates among IDUs between low- and high-HIV-prevalence regions in the United States. The regional difference in HIV seropositivity rate was decomposed into different components: (1) a "rate effect," which was attributed to the differences in factor-specific rates, and (2) "compositional factor effects," which were attributed to the differences in distributions of sociodemographic factors across regions. The analytic results show that the regional difference in HIV seropositivity rate was considerable (21.04%); however, the difference would be adjusted down to 17.65% if sociodemographic factors were proportionally distributed across the regions. Differential distribution of ethnic groups between the two regions accounted for about 15.02% of the regional difference in HIV seropositivity rate. The application of the standardization and decomposition method provides HIV researchers with opportunities to look at familiar data from a different perspective. PMID- 14534386 TI - Sexual health risks among young Thai women: implications for HIV/STD prevention and contraception. AB - This paper examines factors that may place female Thai adolescents and young adults at risk for HIV, sexually transmitted diseases (STDs), and unintended pregnancies. A total of 832 female vocational students participated in a cross sectional audio-computer-assisted self-interview (ACASI) survey after providing informed consent. The questionnaire covered: sociodemographic characteristics; knowledge, attitudes, and beliefs related to HIV and STDs; contraceptive practices; sexual experiences and behaviors; and drug use. Oral fluid was tested for HIV antibodies and urine was tested for illicit drugs and for the presence of gonococcal or chlamydial nucleic acids. A total of 359 women (43.1%) reported sexual intercourse history, with an average age at first sex of 17.6 years, and a 2.6 mean number of lifetime sex partners. Twenty-one percent of the entire sample reported coerced sexual contact or intercourse. Among those with sexual intercourse experience, 27.3% (n=98) had been pregnant and the majority of their most recent pregnancies were terminated. Three tested positive for HIV antibodies. Sexually active young Thai women report behaviors or experiences that may expose them to HIV/STD infection and unintended pregnancy in the future. These include unprotected intercourse, sexual coercion, low levels of contraceptive use, and drug and alcohol use. Culturally appropriate interventions that increase their awareness of and ability to respond to these sexual health risks are needed. PMID- 14534387 TI - A Guttman scale for assessing condom use skills among college students. AB - HIV risk-reduction interventions frequently teach correct condom use skills, yet few researchers actually measure those skills. Without measuring condom use skills, it is not possible to evaluate the effectiveness of condom use interventions, or determine what variables may be related to those skills. The Measure of Observed Condom Use Skills (MOCUS) was developed using standard Guttman scaling procedures, and administered to 106 female and 71 male undergraduates. The MOCUS was administered by the primary investigator to each participant individually. Guttman scale analyses demonstrated the MOCUS has acceptable reproducibility. These findings support future use of the MOCUS to assess condom use skills. PMID- 14534388 TI - HIV risk behaviors and their correlates among HIV-positive adults with serious mental illness. AB - HIV risk behaviors and their correlates were examined in a sample of 154 HIV seropositive and seriously mentally ill adults (83% male, 56% sexually active). Most sexually active participants engaged in HIV risk behavior during the past 6 months, although the rates of these behaviors were generally not higher, and in some cases were lower, than those reported in studies of otherwise comparable noninfected people. Variables significantly associated with one or more HIV risk behaviors in bivariate analyses included being female, any limitation in instrumental functioning, not having a bipolar disorder, more psychotic mental health symptoms, problem drinking, and not receiving HIV counseling. The latter three variables accounted for 22% of the variance in the total number of HIV risk behaviors. Interventions to reduce risky sexual practices may be most appropriately provided through public mental health systems, given that this is the primary setting in which seriously mentally ill adults receive formal treatment. PMID- 14534389 TI - Stress and depression among HIV-positive and HIV-negative gay and bisexual AIDS caregivers. AB - From a survey of 416 gay and bisexual male AIDS caregivers, we examined differences in the stress process and predictors of depressive symptomatology by caregivers' HIV serostatus (n=164 HIV-positive and n=252 HIV-negative). Results indicate that HIV-positive caregivers reported higher levels of depressive symptomatology than those who were HIV-negative. Parallel regression analyses revealed that a younger age, role-related stress, and low self-esteem were common predictors of depressive symptomatology for both groups. Group-specific predictors of depression for the HIV-positive caregivers were poorer health and financial concerns. In contrast, the group-specific predictor for the HIV negative caregivers was higher stress associated with direct acts of caregiving. These results suggest that in order to stem depressive symptomatology, gay and bisexual male caregivers need support directed at reducing stress stemming from their role as caregivers as well as services designed to address low self-esteem. In addition, serostatus should be taken into account when designing service plans because seropositive and seronegative caregivers may require different supportive services. PMID- 14534390 TI - Situational correlates of condom use in a sample of African-American drug users who are primarily crack cocaine users. AB - We studied the situational determinants of condom use for vaginal sex in 151 African-American crack cocaine users in Houston, Texas, using situational presentation (Sitpres) methodology, which uses hypothetical scenarios with randomly generated levels of eight variables across 10 scenarios that may impact the decision to use a condom. Multiple regression showed that variables associated with a high probability of using a condom were older age, how badly the partner wants to use a condom, and how badly the respondent wants vaginal sex. Crack craving or level of intoxication were not correlates. The Sitpres methodology was successfully used by these crack users and data indicated that it was sexual variables, rather than drug-associated variables, that were significantly associated with condom use. PMID- 14534391 TI - Adherence to medication treatment: a qualitative study of facilitators and barriers among a diverse sample of HIV+ men and women in four US cities. AB - Most studies examining HIV antiretroviral medication treatment adherence involve quantitative surveys. Although these studies have identified factors associated with medical adherence, no single variable or combination of variables is sufficiently consistent to apply to any individual or group of people. Using qualitative methods, an ethnically diverse sample (N=110) of HIV+ women, men who have sex with men, and male injecting drug users in four U.S. cities were interviewed in depth to elicit their experiences, perspectives, and life contexts regarding knowledge, attitudes, beliefs, and experiences with HIV medication adherence. Most described multiple influences on medication-taking behavior, describing adherence as a dynamic phenomenon that changes over time with their changing beliefs, attitudes, emotions, and daily and larger life events. Prevalent themes include ambivalence toward HIV medication and intentional nonadherence, usually to address physical side effects. Factors from different domains (e.g., cognitive, emotional, interpersonal) can have compensatory influences on behavioral outcomes. Findings are discussed in terms of social action theory, contributing to our theoretical understanding of the phenomenon of adherence. PMID- 14534392 TI - HIV risk reduction among African-American women who inject drugs: a randomized controlled trial. AB - A community-based HIV intervention for African-American women who are active injection drug users (IDUs) was evaluated. Seventy-one women (aged 20-54 years) were randomly assigned to one of two enhanced gender- and culturally specific intervention conditions or to the NIDA standard condition. Substantial decreases (p<.001) were found in the frequency of drug use and the frequency of drug injection as well as in the sharing of injection works or water and the number of injections. Trading sex for drugs or money, having sex while high, as well as other sexual risk behaviors were also reduced significantly. Furthermore, women in both enhanced intervention conditions were more likely to reduce their drug using and sexual risk behaviors than were women in the standard condition. Results indicate the value of including additional components in interventions designed to reduce the risk of infection with HIV among women who inject drugs. PMID- 14534393 TI - A qualitative investigation of antiretroviral therapy among injection drug users. AB - Recent advances in antiretroviral therapy for HIV/AIDS have improved the quality of life and life expectancies of many with this fatal disease. Nevertheless, a significant proportion of individuals from disadvantaged groups, which traditionally have had difficulty accessing high-quality health care in the United States, have not benefited from these treatments. For example, injection drug users (IDUs), now a principal source of new cases of AIDS, have received antiretroviral therapy at significantly lower rates than other groups. Whereas numerous studies have described this group quantitatively, few studies have examined in depth the influences that lead to IDUs' problematic relationship with HIV/AIDS treatment. The study described here is based on a longitudinal survey of 516 IDUs. A quantitative analysis was conducted with 103 HIV-positive IDUs and an in-depth qualitative analysis was performed with a subsample of 34 HIV-positive IDUs. This paper presents findings on the consequences of antiretroviral therapy use and nonuse among these individuals, focusing on their perspectives regarding this treatment regimen. Findings suggest that problematic access to, and usage of, antiretroviral therapy is related to wider societal problems of drug abuse, access to health care and housing, and the social and historical divisions that impact the lives of IDUs and their communities. Efforts to improve the availability and the adherence to antiretroviral therapy among IDUs should involve these individuals as a critical component in the design of culturally appropriate and supportive health care services. PMID- 14534394 TI - Sleep disorders in patients with congestive heart failure. AB - PURPOSE OF REVIEW: This review of recent literature pertains to the growing evidence that obstructive sleep apnea contributes to the development of systemic hypertension and congestive heart failure. RECENT FINDINGS: There is irrefutable evidence that OSA causes systemic hypertension and that continuous positive airway pressure (CPAP) treatment of OSA causes a reduction in blood pressure. Moreover there is evidence that untreated OSA is associated with left ventricular diastolic and systolic failure and that treatment with CPAP improves systolic function. SUMMARY: OSA should be considered in patients with systemic hypertension or heart failure. PMID- 14534395 TI - Recent advances in understanding the pathogenesis of obstructive sleep apnea. AB - PURPOSE OF REVIEW: The pathogenesis of obstructive sleep apnea (OSA) is incompletely understood. Historically it was believed that patients with OSA have a small upper airway (often due to obesity) that is kept patent during wakefulness by the activity of upper airway dilating muscles. With the reduction in muscle tone at sleep onset, the airway collapses and causes apnea. While this appears to be the case for many patients with OSA, other patients show no major airway anatomic defects or minimal obesity. RECENT FINDINGS: This has led to the concept that other factors such as unstable ventilatory control and changes in lung volume during sleep may be involved in the pathogenesis of OSA. Recently there have been several advances in our understanding of how these mechanisms are involved in OSA pathogenesis. SUMMARY: A more complete understanding of apnea pathogenesis may improve therapeutic techniques and reduce the consequences of OSA. PMID- 14534396 TI - Role of portable sleep studies for diagnosis of obstructive sleep apnea. AB - PURPOSE OF REVIEW: There is growing awareness of the significance of obstructive sleep apnea in the general population and in the medical community and, as a result, there is a growing demand for diagnosis and treatment. Attended, in laboratory polysomnography is resource intensive and not readily available in some communities. Alternate diagnostic strategies have been proposed including the use of home sleep studies. Although these portable systems have been in use for many years, only in the past few years have a significant number of studies been performed to evaluate these systems in the home setting. The use of actigraphy and peripheral arterial tonometry for diagnostic purposes has also recently been investigated. RECENT FINDINGS: In the laboratory setting, measurements of sleep-disordered breathing with specific portable sleep systems correspond well with measurements provided by standard polysomnography. In the home setting, portable systems demonstrate several important limitations including lost or inadequate data collection, logistic concerns, and mildly reduced diagnostic accuracy. Data regarding the potential cost benefit of home studies is inconclusive. SUMMARY: Home polysomnography is a viable option for evaluating patients with moderate or high clinical suspicion for sleep-disordered breathing. However, patients with failed or equivocal home studies and those with negative studies but persistent symptoms should undergo standard polysomnography. Further investigations are needed to compare long-term outcomes in patients evaluated using portable devices versus standard polysomnography. PMID- 14534397 TI - Sleep disorders in children. AB - PURPOSE OF REVIEW: Childhood sleep disorders are one of the most prevalent complaints in the pediatrician's office. Infant sleep rhythm complaints from new mothers reach 46%, while childhood obstructive sleep apnea has a prevalence of 2% and adolescent insomnia with daily consequences surpasses that percentage. RECENT FINDINGS: Each sleep disorder must be considered in context of age, as age influences the presentation and impact on the developing child or adolescent. For example, sleep-disordered breathing resulting in adult sleepiness can contribute to death in infants. The symptoms of narcolepsy are often masked until after adolescence, resulting in psychologically costly misdiagnoses. SUMMARY: There are no outcome studies that track the long-term consequences of pediatric sleep disorders or their contribution to adult sleep problems, but this is an area of increasing research interest. This review assesses the most recent literature on pediatric sleep disorders from May 1, 2002, until April 30, 2003. PMID- 14534398 TI - Sleep disorders in pregnancy. AB - PURPOSE OF REVIEW: Sleep disturbances are frequent during pregnancy. The spectrum of association between pregnancy and sleep disturbances ranges from an increased incidence of insomnia, nocturnal awakenings, and parasomnias (especially restless legs syndrome) to snoring and excessive sleepiness. These disturbances occur as a result of physiologic, hormonal, and physical changes associated with pregnancy. Although the timing and occurrence of different sleep disorders varies, they are most prevalent during the third trimester. Despite reports of the various sleep problems, the exact nature and incidence of sleep disorders in pregnancy is not known. Given that limitation, we are presenting an up-to-date review of the current understanding of the relation between sleep and pregnancy. RECENT FINDINGS: Studies suggest that pregnancy affects sleep in multiple ways. There are hormonal changes, physiologic changes, physical factors, and behavioral changes in a pregnant woman-all of which may affect her sleep. They may affect the duration and quality of sleep and lead to a variety of sleep disorders. Pregnancy may also affect an existing sleep disorder. Particular attention may be given to obese pregnant women who would gain more weight during pregnancy or those who develop hypertensive conditions (eg, preeclampsia). Snoring may be more common in women with preeclampsia and the pressor responses to obstructive respiratory events during sleep may be enhanced in preeclamptic women when compared with those with obstructive sleep apnea alone. Several investigators have suggested that obstructive sleep apnea (OSA) may be common in pregnant women despite the presence of intrinsic mechanisms that seem to be geared towards preventing sleep apnea. However, the exact incidence and prevalence of sleep apnea in pregnant women is uncertain. In addition, it is unclear if criteria that are used to define sleep apnea in the general population should be applied to pregnant women. Further investigations are needed to determine if lower thresholds for management of OSA should be used in pregnant women to prevent harm to the fetus. SUMMARY: In conclusion, sleep disturbances are common during pregnancy though the full extent of this relation remains undefined. Large, multi center, prospective studies are needed for better understanding. PMID- 14534399 TI - Cystic fibrosis. PMID- 14534400 TI - Update on pathogenesis of cystic fibrosis lung disease. AB - PURPOSE OF REVIEW: It has been an ongoing challenge to translate knowledge pertaining to the molecular basis of cystic fibrosis (CF) into a clear understanding of the development of CF lung disease. Various hypotheses have attempted to explain the apparent breach of innate defenses in CF, although a definitive explanation has been elusive. RECENT FINDINGS: Recent data suggest that altered ion transport functions--namely sodium hyperabsorption and reduced chloride secretion--lead to a depletion of airway surface liquid. As a result, the overlying mucus layer may encroach upon cell surfaces and become adherent, thus interfering with cilia-dependent and cough clearance. These static, and ultimately anaerobic, niches provide a favorable environment for the development of bacterial biofilms and persistent infection with Pseudomonas aeruginosa. SUMMARY: With a better understanding of pathogenic steps leading to CF lung disease, we may now be able to direct the development of therapies that will substantially improve disease outcomes. PMID- 14534401 TI - Pseudomonas acquisition in young patients with cystic fibrosis: pathophysiology, diagnosis, and management. AB - PURPOSE OF REVIEW: To summarize the pathophysiology of, risk factors for, and outcomes of early Pseudomonas aeruginosa (Pa) infection in CF; to review the results of trials of early intervention and to describe treatment options for early Pa infection. RECENT FINDINGS: Chronic lower airway Pa infection is associated with significant morbidity and mortality among CF patients. However, first acquisition of Pa does not appear to cause an immediate and rapid decline in lung function. Early Pa isolates are generally non-mucoid, antibiotic sensitive, and present at low density, suggesting a possible "window of opportunity" for early intervention. SUMMARY: Anti-pseudomonal therapy for early infection results in transient Pa eradication, but re-infection with Pa appears inevitable despite early aggressive treatment. There are no controlled trials demonstrating clinical benefit in young children. There is a critical need for further investigation of the clinical outcomes associated with early intervention, the long-term safety profile, and the optimal drug regimen. PMID- 14534403 TI - Long-acting bronchodilators in cystic fibrosis. AB - PURPOSE OF REVIEW: Over 80% of patients with cystic fibrosis (CF) have bronchodilator therapy prescribed, yet bronchodilator use in CF remains controversial. The development of long-acting beta-agonist drugs for clinical use has provided additional rationale for considering bronchodilator therapy in CF. This paper will review recent developments in bronchodilator use in CF patients, with emphasis on the long-acting beta agonists. RECENT FINDINGS: It is reported that 50 to 60% of CF patients demonstrate significant intermittent airway hyperreactivity in response to bronchodilators or challenges. The beta-agonist drugs are the most commonly prescribed bronchodilators. Several mechanisms may be implicated in therapeutic response of CF patients to bronchodilators including direct smooth muscle relaxation, increased mucociliary clearance, direct effects on inflammatory cells and bacterial adherence, and possible direct effects on CF transmembrane conductance regulator (CFTR) function. Several recent studies have shown improved outcomes with long-acting bronchodilators. SUMMARY: In spite of the widespread use of bronchodilators, there are very few long-term studies of their effects in CF patients. However, there are clearly clinical benefits in certain situations. Further research into the most appropriate utilization of these medications to improve outcomes in patients with CF would be helpful. PMID- 14534402 TI - Nonclassic cystic fibrosis and CFTR-related diseases. AB - PURPOSE OF REVIEW: To review the spectrum of disease caused by mutations in the cystic fibrosis (CF) gene. RECENT FINDINGS: The growing recognition of "atypical" cases of cystic fibrosis presenting in adolescence or adulthood and manifested by disease in only one or two organ systems, along with CF diagnostic criteria based not only on sweat chloride values but genetic screening and nasal ion transport measurements, have made the diagnosis of CF less straightforward for many clinicians. SUMMARY: This review seeks to clarify the key diagnostic criteria for CF and uses the Cystic Fibrosis Foundation's Consensus Diagnostic Guidelines and recent publications to discuss the characteristics of classic CF, nonclassic CF, and CFTR-related diseases. PMID- 14534404 TI - Approach to resistant gram-negative bacterial pulmonary infections in patients with cystic fibrosis. AB - PURPOSE OF THE REVIEW: Patients with cystic fibrosis are living longer with chronic pulmonary bacterial infections. One consequence of antibiotic treatment of these chronic infections has been the increasing prevalence of antibiotic resistance seen in bacterial isolates recovered from patients with cystic fibrosis. RECENT FINDINGS: Bacteria such as Pseudomonas aeruginosa and Burkholderia cepacia are able to acquire antibiotic resistance by either spontaneous mutation or gene transfer via plasmids or integrins. In addition, bacteria survive by forming antibiotic-resistant biofilms within the airways of patients with cystic fibrosis. Therapeutic approaches to dealing with antibiotic resistant bacterial pulmonary infections include the use of in vitro synergy testing to determine optimal double antibiotic combinations or multiple combination bactericidal testing to determine bactericidal double and triple antibiotic combinations to use against the bacteria in the clinical setting of acute exacerbations. SUMMARY: Therapy for antibiotic-resistant bacterial infections in cystic fibrosis involves the use of new laboratory methods (synergy testing or multiple-combination bactericidal testing) to optimize antibiotic treatment strategies. Clinical trials are required to address whether treatment guided by susceptibility testing improves clinical outcomes. Future novel approaches will likely include drugs that can disrupt bacterial biofilm formation and the use of cationic peptide antimicrobial compounds. PMID- 14534405 TI - Noninvasive biomarkers of airway inflammation in cystic fibrosis. AB - PURPOSE OF REVIEW: Airway inflammation plays a central role in the lung disease of cystic fibrosis (CF). Biomarkers of inflammation may be useful for monitoring disease progression and evaluating response to therapy. Much of our knowledge of the chronic inflammatory process in the CF airway derives from studies of bronchoscopy and bronchoalveolar lavage. A number of noninvasive approaches have been recently developed to more readily assess airway inflammation including sputum induction, collection of exhaled air, analysis of systemic markers of inflammation, and computed tomography imaging. RECENT FINDINGS: While measurements of biomarkers of inflammation continue to advance our understanding of the underlying disease process, there is as yet no established role for these markers in clinical practice. This review summarizes the current state of knowledge of various inflammatory markers relevant to CF lung disease, with an eye towards application as surrogate outcome measures in CF clinical trials. SUMMARY: It is hoped that biomarkers obtained by noninvasive means will be useful in determining specific pathways of injury (ie, oxidative or proteolytic) in individual persons with CF and in assessing response to antiinflammatory treatments. PMID- 14534407 TI - Cochlear compression: perceptual measures and implications for normal and impaired hearing. AB - This article provides a review of recent developments in our understanding of how cochlear nonlinearity affects sound perception and how a loss of the nonlinearity associated with cochlear hearing impairment changes the way sounds are perceived. The response of the healthy mammalian basilar membrane (BM) to sound is sharply tuned, highly nonlinear, and compressive. Damage to the outer hair cells (OHCs) results in changes to all three attributes: in the case of total OHC loss, the response of the BM becomes broadly tuned and linear. Many of the differences in auditory perception and performance between normal-hearing and hearing-impaired listeners can be explained in terms of these changes in BM response. Effects that can be accounted for in this way include poorer audiometric thresholds, loudness recruitment, reduced frequency selectivity, and changes in apparent temporal processing. All these effects can influence the ability of hearing-impaired listeners to perceive speech, especially in complex acoustic backgrounds. A number of behavioral methods have been proposed to estimate cochlear nonlinearity in individual listeners. By separating the effects of cochlear nonlinearity from other aspects of hearing impairment, such methods may contribute towards identifying the different physiological mechanisms responsible for hearing loss in individual patients. This in turn may lead to more accurate diagnoses and more effective hearing-aid fitting for individual patients. A remaining challenge is to devise a behavioral measure that is sufficiently accurate and efficient to be used in a clinical setting. PMID- 14534409 TI - A software tool for analyzing multichannel cochlear implant signals. AB - A useful and convenient means to analyze the radio frequency (RF) signals being sent by a speech processor to a cochlear implant would be to actually capture and display them with appropriate software. This is particularly useful for development or diagnostic purposes. sCILab (Swiss Cochlear Implant Laboratory) is such a PC-based software tool intended for the Nucleus family of Multichannel Cochlear Implants. Its graphical user interface provides a convenient and intuitive means for visualizing and analyzing the signals encoding speech information. Both numerical and graphic displays are available for detailed examination of the captured CI signals, as well as an acoustic simulation of these CI signals. sCILab has been used in the design and verification of new speech coding strategies, and has also been applied as an analytical tool in studies of how different parameter settings of existing speech coding strategies affect speech perception. As a diagnostic tool, it is also useful for troubleshooting problems with the external equipment of the cochlear implant systems. PMID- 14534410 TI - Effects of presentation level on phoneme and sentence recognition in quiet by cochlear implant listeners. AB - OBJECTIVE: The objectives of this study were to characterize the effects of presentation level on speech recognition in quiet by cochlear implant users with the Nucleus 22 SPEAK and Clarion v1.2 CIS speech-processing strategies, and to relate speech recognition at low presentation levels to stimulus audibility as measured by sound field thresholds. It was hypothesized that speech recognition performance in both Nucleus SPEAK and Clarion CIS participants would decrease as presentation level was decreased below 50 to 60 dBA, due to audibility limitations. However, it was expected that such level effects would be less severe in CIS participants than in SPEAK participants because the Clarion v1.2 device encodes a wider acoustic dynamic range (up to 60 dB) than the Nucleus 22 device (30 dB). DESIGN: Performance-intensity (P-I) functions for vowels, consonants and sentences in quiet were obtained from each participant. P-I functions incorporated speech levels of 70, 60, 50, 40 and 30 dBA. Subjects used their clinical speech processor maps and adjusted the loudness (volume/sensitivity) controls on their processors so that speech presented at 60 dBA was comfortably loud. Maps were created using default clinical procedures and were not adjusted to optimize sound field thresholds. Sound field thresholds and dynamic ranges were measured for warbled pure tones with frequencies of 250 to 6000 Hz. RESULTS: Consonant and sentence recognition showed strong level effects for both SPEAK and CIS participants, with performance decreasing substantially at levels below 50 dBA in most individuals. Vowel recognition showed weaker level effects. For all three speech materials, SPEAK and CIS participants demonstrated similar mean performance at 70 dBA; however, SPEAK participants showed larger reductions in performance than CIS participants with decreasing level. Sound field thresholds were more sensitive for CIS participants than for SPEAK participants, supporting the hypothesis that performance differences were related to audibility. CONCLUSIONS: Cochlear implant listeners are unable to maintain good speech recognition at low presentation levels due to reduced stimulus audibility, and this may significantly limit their ability to communicate in daily life. It is likely that audibility differences between SPEAK and CIS participants in the present study can be attributed at least partly to differences in the acoustic dynamic range used by the respective processors. However, several additional factors may have contributed to differences in audibility and perception of soft speech among individual listeners with both devices. These include the minimum and maximum electrical stimulation levels specified in participants' maps and the speech processor sensitivity setting used for testing. PMID- 14534408 TI - Sources and mechanisms of DPOAE generation: implications for the prediction of auditory sensitivity. AB - Otoacoustic emissions (OAEs) have become a commonly used clinical tool for assessing cochlear health status, in particular, the integrity of the cochlear amplifier or motor component of cochlear function. Predicting hearing thresholds from OAEs, however, remains a research challenge. Models and experimental data suggest that there are two mechanisms involved in the generation of OAEs. For distortion product, transient, and high-level stimulus frequency emissions, the interaction of multiple sources of emissions in the cochlea leads to amplitude variation in the composite ear canal signal. Multiple sources of emissions complicate simple correlations between audiometric test frequencies and otoacoustic emission frequencies. Current research offers new methods for estimating the individual components of OAE generation. Input-output functions and DP-grams of the nonlinear component of the 2f2-f2 DPOAE may ultimately show better correlations with hearing thresholds. This paper reviews models of OAE generation and methods for estimating the contribution of source components to the composite emission that is recorded in the ear canal. The clinical implications of multiple source components are discussed. PMID- 14534411 TI - Efficient stimuli for evoking auditory steady-state responses. AB - OBJECTIVE: To compare the magnitudes of the steady-state responses evoked by several types of stimuli, and the times required to recognize these responses as significant. DESIGN: In the first two experiments, we examined auditory steady state responses to pure tones, broadband noise and band-limited noise. The stimuli were amplitude modulated in the 75 to 100 Hz range with sinusoidal or exponential envelopes. A third experiment investigated the effects of exponential envelopes on the responses to broadband noise. The final experiment examined auditory steady-state responses evoked by rapidly presented transient stimuli, such as clicks, brief tones and brief noise-bursts. All stimuli were presented dichotically at intensities 30 to 50 dB above behavioral thresholds. The subjects were adults, who drowsed or slept during the recording sessions. RESULTS: The responses to the noise were larger than the responses to the tones. At an intensity of 32 dB nHL, the average amount of time needed to obtain significant responses for the amplitude-modulated noise was 43 sec and the maximum time was 2 minutes. The average time for pure tone stimuli was approximately 2 minutes but 25% of the responses remained undetected after 5 minutes. Combining the responses to all the frequency-specific stimuli showed results similar to using noise stimuli. Using exponential envelopes did not increase response amplitudes for noise stimuli. At 45 dB nHL, the steady-state responses to clicks and other transient stimuli were larger than responses to the broadband noise. The average time to detect steady-state responses to transient stimuli was approximately 20 sec, which was a little faster than for amplitude modulated noise. CONCLUSIONS: Auditory steady-state potentials evoked by amplitude modulated noise or transient stimuli might be useful in providing rapid and objective tests of hearing during screening procedures. Another approach might be to record responses to multiple frequency-specific stimuli and to evaluate the combined responses for a rapid indication that some hearing is present. PMID- 14534412 TI - Full time directional versus user selectable microphone modes in hearing aids. AB - OBJECTIVE: The purpose of this experiment was to systematically examine hearing aid benefit as measured by speech recognition and self-assessment methods across omnidirectional and directional hearing aid modes. These data were used to compare directional benefit as measured by speech recognition in the laboratory to hearing aid wearer's perceptions of benefit in everyday environments across full-time directional, full-time omnidirectional, and user selectable directional fittings. Identification of possible listening situations that resulted in different self reported hearing aid benefit as a function of microphone type was a secondary objective of this experiment. DESIGN: Fifteen adults with symmetrical, sloping sensorineural hearing loss were fitted bilaterally with in the-ear (ITE) directional hearing aids. Measures of hearing aid benefit included the Profile of Hearing Aid Benefit (PHAB), the Connected Sentence Test (CST), the Hearing in Noise Test (HINT), and a daily use log. Additionally, two new subscales were developed for administration with the PHAB. These subscales were developed to specifically address situations in which directional hearing aids may provide different degrees of benefit than omnidirectional hearing aids. Participants completed these measures in three conditions: omnidirectional only (O), directional only with low-frequency gain compensation (D), and user selectable directional/omnidirectional (DO). RESULTS: Results from the speech intelligibility in noise testing indicated significantly more hearing aid benefit in directional modes than omnidirectional. PHAB results indicated more benefit on the background noise subscale (BN) in the DO condition than in the O condition; however, this directional advantage was not present for the D condition. Although the reliability of the newly proposed subscales is as yet unknown, the data were interpreted as revealing a directional advantage in situations where the signal of interest was in front of the participant and a directional disadvantage in situations where the signal of interest was behind the listener or localization was required. CONCLUSIONS: Laboratory directional benefit is reflected in self assessment measures that focus on listening in noise when the sound source of interest is in front of the listener. The use of a directional hearing aid mode; however, may have either a positive, a neutral, or a negative impact on hearing aid benefit measured in noisy situations, depending on the specific listening situation. PMID- 14534413 TI - GJB2 mutations in the Swiss hearing impaired. AB - OBJECTIVE: Mutations in the GJB2 gene encoding connexin 26 (Cx26) protein are a major cause for nonsyndromic autosomal recessive and sporadic deafness. However, its contribution to hearing impairment in Switzerland remains undefined. To determine the frequency and type of GJB2 mutations in the Swiss hearing-impaired population diagnosed under the age of 2 yr and at 2 yr and older and to assess the effectiveness of denaturing high-performance liquid chromatography (DHPLC) in screening for mutation in GJB2. METHODS: Thirty-four patients with hearing impairment underwent mutation screening of the single coding exon of GJB2 with DHPLC followed by bidirectional sequencing to identify sequence alterations. RESULTS: GJB2 mutations were more common in children diagnosed with hearing impairment under the age of 2 yr compared to the group 2 yr and older. In patients under age 2 yr, 9 of 20 (45%) harbored 13 GJB2 mutations including a common 313del14nt mutation; four of these patients were homozygous or compound heterozygous for GJB2 mutations. In contrast, 2 of 14 patients in the 2 yr and older group (14%) had a single mutation in GJB2. The 35delG mutation was exclusively found in 5 patients under the age of 2 yr. DHPLC for mutation screening was 100% sensitive and 83% specific for detecting sequence alterations in GJB2. CONCLUSIONS: In Switzerland, GJB2 mutations are a major cause of nonsyndromic hearing impairment in children under the age of 2. Similar to other populations, GJB2 mutations are uncommon in the affected Swiss patients identified after 2 yr. Although 35delG mutation is common in the hearing-impaired children under the age of 2, it was absent in patients diagnosed with hearing impairment after the age of 2. DHPLC is a highly sensitive tool for detection of GJB2 mutations. PMID- 14534414 TI - Comorbid auditory processing disorder in developmental dyslexia. AB - OBJECTIVE: The primary objective of this study was to investigate the extent of comorbid auditory processing disorder (APD) in a group of adults with developmental dyslexia. An additional objective was to compare performance on auditory tasks to results from standardized tests of reading in an attempt to generate a clinically useful profile of developmental dyslexics with comorbid APD. DESIGN: A group of eleven persons with developmental dyslexia and 14 age- and intelligence-matched controls participated in the study. Behavioral audiograms, 226-Hz tympanograms, and word recognition scores were obtained binaurally from all subjects. Both groups were administered the frequency-pattern test (FPT) and duration-pattern test (DPT) monaurally (30 items per ear) in both the left and right ear. Gap detection results were obtained in both groups (binaural presentation) using narrowband noise centered at 1 kHz in an adaptive two-alternative forced-choice (2-AFC) paradigm. The FPT, DPT, and gap detection results were analyzed for interaural (where applicable), intergroup, and intragroup differences. Correlations between performance on the auditory tasks and the standardized tests of reading were examined. Additive logistic regression models were fit to the data to determine which auditory tests proved to be the best predictors of group membership. RESULTS: The persons with developmental dyslexia as a group performed significantly poorer than controls on both the FPT and DPT. Furthermore, the group differences were significant in both monaural conditions. On the FPT and DPT, five of the eleven participants with dyslexia performed below the widely used clinical criterion for APD of 70% correct in either ear. All five of these participants performed below criterion on the FPT, whereas four of the five additionally performed below 70% on the DPT. The data also were analyzed by fitting a series of stepwise logistic regression models, which indicated that gap detection did not significantly predict group membership, whereas the FPT and DPT were significant predictors. The addition of the FPT score after the DPT did not result in a significant change in the residual deviance. CONCLUSIONS: Approximately half of the participants with developmental dyslexia showed clinically significant diminished performance on the FPT and DPT indicative of APD. These results indicate that the percentage of persons with developmental dyslexia and comorbid APD may be substantial enough to warrant serious clinical considerations. PMID- 14534415 TI - Adaptation by a cochlear-implant patient to upward shifts in the frequency representation of speech. AB - The purpose of this project was to assess the degree to which a patient, after 1 wk of experience, could adapt to 3.2-mm and 6.8-mm basal shifts in the representation of speech. Only small deficits in performance were found after practice after the 3.2-mm shift. After practice after the 6.9-mm shift, scores on tests that emphasized amplitude envelope cues returned to baseline levels. Scores on vowel and sentence tests that emphasized frequency-based cues remained poor. Scores for "place," however, showed some recovery. Vowel recognition may be the limiting factor in recognizing basally shifted speech. PMID- 14534417 TI - The neural computation of the aperture problem: an iterative process. AB - The aperture problem is defined as one of integrating motion information from inside and outside of the aperture, and determination of the true direction of motion of a line. Much is known about it and many models have been proposed for its neural mechanisms. However, it is still a matter of debate whether the brain solves the problem by using only feed-forward neural connections, also known as the one-shot algorithm, or by using the iterative algorithm while utilizing feedback as well as horizontal neural connections. Here we show unequivocal evidence for the latter model. The model was tested using critically designed psychophysical experiments and the results were perfectly in line with the psychophysical performance of the observers. PMID- 14534416 TI - Voltage-gated K(+) channel subunits on cholinergic and dopaminergic amacrine cells. AB - The expression of voltage-gated K+ (Kv) channel subunits on rat retinal cholinergic and dopaminergic amacrine cells was studied using double immunofluorescence labeling and confocal laser scanning microscopy. Staining for Kv3.1b was found in the cholinergic cells, being present on the membrane of somata, and on the processes, but not in the dopaminergic cells. Kv4.3 immunoreactivity was localized on the somatodendritic compartment of the dopaminergic cells, but was not found in the cholinergic cells. Differential expression between the two cell types was not found for 10 other subunits tested. These results suggest that the Kv3.1b and Kv4.3 subunits may differentially contribute to the electrophysiological properties underlying distinct functions of these two retinal interneurons. PMID- 14534418 TI - ERP abnormalities of illusory contour perception in Williams syndrome. AB - Williams syndrome is a genetic disorder in which visuo-spatial performance is poor. Theorists have claimed that the deficit lies in high-level processing, leaving low-level visual processes intact. We investigated this claim by examining an aspect of low-level processing, perceptual completion, i.e. the ability of this clinical group to perceive illusory Kanizsa squares. We then used event-related potentials to examine neural correlates of perceptual completion. While participants were able to perceive illusory contours, the neural correlates of this apparently normal perception were different from controls. Such differences in low-level visual processes may significantly impact on the development of higher-level visual processes. We conclude that, contrary to earlier claims, there is atypical neural processing during low-level visual perception in Williams syndrome. PMID- 14534419 TI - Lithium inhibits apoptosis of mouse neural progenitor cells. AB - As undifferentiated precursor cells in the CNS, neural progenitor cells (NPCs) supply new neurons and glial cells to repair damage within the adult brain. Recently, NPCs were found to undergo apoptosis. In serum-free basic fibroblast growth factor-containing culture medium, primary culture cells from fetal mouse neuroepithelium expressed nestin, and thus might be regarded as NPCs. These NPCs demonstrated apoptosis under electron microscopy and TUNEL assay. Treatment of NPCs with lithium, a specific inhibitor of glycogen synthase kinase 3beta (GSK3beta), significantly suppressed apoptosis. Activity of pro-apoptotic protease caspase-3 was not detected in either lithium-untreated or -treated NPCs. These findings suggest that lithium may protect NPCs against apoptosis by inhibiting caspase-3-independent apoptotic pathways. PMID- 14534420 TI - Reciprocal accumulation of beta-synuclein in alpha-synuclein lesions in multiple system atrophy. AB - Alpha-Synuclein is a major component of neuronal and glial cytoplasmic inclusions in multiple system atrophy (MSA), one of the alpha-synucleinopathies. Recent studies have shown that beta-synuclein, a homolog of alpha-synuclein, inhibits alpha-synuclein aggregation in vitro. We immunohistochemically examined the MSA brain, using specific antibodies against alpha-synuclein and beta-synuclein. alpha-synuclein-positive filamentous aggregates were frequently found in neurons in the pontine and inferior olivary nuclei. No abnormal accumulation of alpha synuclein was noted in Purkinje cells. In contrast, beta-synuclein accumulation occurred extensively in Purkinje cells, and only minimally in pontine and olivary neurons. Thus, neuronal alpha-synuclein inclusions appear to occur only rarely in neurons in which beta-synuclein accumulates. These findings support the possibility that beta-synuclein is a negative regulator of alpha-synuclein aggregation. PMID- 14534421 TI - Sensory deprivation changes the pattern of synaptic connectivity in rat barrel cortex. AB - We examined whether sensory deprivation during formation of the cortical circuitry influences the pattern of intracortical single-cell connections in rat barrel cortex. Excitatory postsynaptic potentials from layer 5 pyramidal neurons were recorded in vitro using patch-clamp techniques. In order to evoke such postsynaptic potentials presumptive presynaptic neurons were stimulated by photolytically applied glutamate thus generating action potentials. Synaptic connections between the stimulated and the recorded neuron were identified by the occurrence of postsynaptic potentials following photostimulation. Sensory deprivation altered the projections from layer 2/3 neurons to layer 5 pyramidal cells (L2/3-->L5 projections). In slices of non-deprived rats the input probability of L2/3-->L5 projections showed a periodic pattern with more synaptic connections originating from the borders of the barrel columns, and less synaptic connections originating from the centres. After whisker clipping this periodic pattern disappeared completely and the input probability declined monotonically with increasing distance between stimulated and recorded neuron. These results indicate that sensory input is a prerequisite to establish a synaptic projection pattern which is correlated to the columnar organisation of the anatomical barrel structure. PMID- 14534422 TI - The effects of stimulus competition and voluntary attention on colour-graphemic synaesthesia. AB - Colour-graphemic synaesthetes experience vivid colours when reading letters, digits and words. We examined the effect of stimulus competition and attention on these unusual colour experiences in 14 synaesthetes and 14 non-synaesthetic controls. Participants named the colour of hierarchical local-global stimuli in which letters at each level elicited synaesthetic colours that were congruent or incongruent with the display colour. Synaesthetes were significantly slower to name display colours when either level was incongruent than when both levels were congruent. This effect was significantly reduced when synaesthetes focused attention on one level while the congruency of letters at the ignored level was varied. These findings suggest that competition between multiple inducers and mechanisms of voluntary attention influence colour-graphemic synaesthesia. PMID- 14534423 TI - Atrophy pattern in SCA2 determined by voxel-based morphometry. AB - We applied voxel-based morphometry, an indirect volumetric technique, to MRI volumes of patients carrying the spinocerebellar ataxia type 2 mutation to determine patterns of brain atrophy. Nine patients were compared to 27 controls matched for age, sex and handedness. An optimised voxel-based morphometry protocol was used for pre-processing to minimize systematic bias. We observed significant volume loss in the cerebellar hemispheres, vermis, pons, mesencephalon and thalamus. Also affected were several supratentorial areas such as the right orbito-frontal cortex, right temporo-mesial cortex and the primary sensorimotor cortex bilaterally. The volumetric changes of cerebellar hemispheres were inversely correlated to cerebellar symptoms rated by a cerebellar ataxia scale. Two mechanisms could contribute to the observed cortical atrophy. It could be either the result of primary supratentorial degeneration as part of the disease process and/or secondary atrophy due to cerebellar deafferentation. PMID- 14534424 TI - Grip force efficiency in long-term deprivation of somatosensory feedback. AB - A subject with complete and chronic absence of cutaneous and proprioceptive feedback applied inefficiently elevated grip forces to lift and hold instrumented objects of varying weight. However, the ability to differentially scale grip forces according to various weights (0.36 kg, 0.61 kg and 0.86 kg) was mainly maintained when the subject was allowed to lift a constant weight repeatedly under constant visual control. The normally very precise temporal coupling between grip and load forces was substantially disturbed for lifts performed by the deafferented subject. In particular, maximum grip force lagging some 100 ms behind maximum load, suggesting rather feedback-based than predictive grip force control. Our findings demonstrate the importance of cutaneous and proprioceptive afferents for predictive regulation of prehensile finger forces. However, alternative sensory signals, e.g. from visual cues, small myelinated and unmyelinated muscle afferents or neck proprioception, may provide some provisional grip force control in the absence of cutaneous and proprioceptive feedback. PMID- 14534425 TI - LTP-induced depression of response to hypoxia in hippocampus: effects of adenosine receptor activation. AB - Previous work has shown that long-term potentiation (LTP) can reduce the effects of hypoxia in depressing population spikes in rat hippocampal slices. We have now investigated the role of adenosine in this phenomenon. There is no mutual inhibition between the depressant effects of hypoxia and adenosine, but LTP reduces responses to both hypoxia and adenosine, as does application of an A1 receptor antagonist. The effect of LTP is not due to a change in the balance of activation of A1 and A2A adenosine receptors since a selective A2A receptor antagonist did not prevent the interaction. We suggest that LTP may reduce the response to hypoxia by attenuating neuronal sensitivity to adenosine A1 receptors. PMID- 14534426 TI - Effect of sesame antioxidants on LPS-induced NO production by BV2 microglial cells. AB - Sesame antioxidants have been shown to inhibit lipid peroxidation and regulate cytokine production. In this study, we focused on the effect of sesamin and sesamolin, on nitric oxide (NO) induction by lipopolysaccharides (LPS) in the murine microglial cell line BV-2 and rat primary microglia. The results showed that sesamin and sesamolin significantly inhibited NO production, iNOS mRNA and protein expression in LPS-stimulated BV-2 cells. Sesamin or sesamolin significantly reduced LPS-activated p38 MAPK of BV-2 cells. Furthermore, SB203580, a specific inhibitor of p38 MAP kinase, dose-dependently inhibited NO production in LPS-stimulated BV-2 cells. Taken together, the inhibition of NO production might be due to the reduction of LPS-induced p38 MAPK signal pathway by sesamin and sesamolin. PMID- 14534427 TI - SRC-1 localisation in lumbosacral spinal cord of male and female Wistar rats. AB - Nuclear receptor co-activators play an important role in enhancing transcriptional activity of steroid hormone receptors, however there is currently little information concerning their distribution within the spinal cord. In this study, the distribution of steroid receptor co-activator-1 (SRC-1) was examined with immunocytochemistry, in the lumbosacral cord of Wistar rats of both sexes. In all rats, regardless of sex, SRC-1 was predominant in neurons of the superficial laminae of the dorsal horn and within motorneurons of lamina IX. Sexually dimorphic nuclei exhibited robust SRC-1 immunoreactivity in young rats, including orchidectomised animals, but this appeared to decline in aged rats. Dorsal horn labelling appeared similarly reduced suggesting a possible age related down-regulation of the transcription mediated by steroid receptors in some spinal neurons. PMID- 14534428 TI - ApoE4 is more efficient than E3 in brain access by herpes simplex virus type 1. AB - Apolipoprotein E (ApoE) plays a relevant role in herpes simplex type 1 (HSV-1) infection of the CNS; after infection by the hematogenous route, the viral neuroinvasiveness directly depends on the APOE gene dose. To analyze the effect of ApoE isoforms on the HSV-1 infectivity to the brain, we have used a model of hematogenous infection of mice humanized for the ApoE3 or the ApoE4 alleles, and we have analyzed the presence of viral DNA in several organs by real time quantitative PCR. We have found that animals expressing human ApoE4 present very high levels of virus in the brain when compared to those expressing the ApoE3 allele; in contrast, there were no significant differences in the viral levels found in peripheral organs. Apolipoprotein E4 seems to facilitate the entry and/or spread of HSV-1 in the brain much more efficiently than E3, pointing to a novel potential mechanism underlying the susceptibility to neurodegenerative processes associated with the ApoE4 allele. PMID- 14534429 TI - Excessive weight gain in rats over extended kindling of the basolateral amygdala. AB - Previous lesion studies have indicated a role of the amygdala in the central regulation of food intake. In the present experiments, twice-daily electrical stimulation of the basolateral nucleus of the amygdala in female Wistar rats was found to be associated with a significant body weight gain compared to unstimulated controls. On average, significant increases in body weight were observed after 25 amygdala stimulations, using a kindling paradigm for stimulation. Compared to kindled rats, in which amygdala stimulations were terminated after about 20 stimulations, extended kindling of the amygdala with up to 280 stimulations led to progressive weight increases and compulsive hyperphagia. No gross neuronal damage was seen in thionin-stained sections of the amygdala after extended kindling, but degeneration of a specific type of neurons can not be excluded. The results substantiate that amygdaloid nuclei are an important extrahypothalamic site for the regulation of food intake and body weight. The extensive weight gain over extended amygdala kindling provides an interesting new model for experimentally induced obesity. PMID- 14534430 TI - Abnormal anterior cingulum in patients with schizophrenia: a diffusion tensor imaging study. AB - Diffusion tensor imaging (DTI) can non-invasively examine the molecular diffusion of water in vivo and directly reflects the anatomical integrity of neural fibers in white matter. Fractional anisotropy (FA) can be calculated from DTI data, and utilized to evaluate white matter integrity. DTI was performed on 30 patients with schizophrenia and 19 healthy controls, and their FA values were subsequently measured in multiple brain regions. Statistical analyses revealed that FA values were decreased in the anterior cingulum of schizophrenia subjects. There were no significant differences between patients and controls in any other regions. This study supports the hypothesis that schizophrenia is associated with abnormal white matter integrity of the anterior cingulum. PMID- 14534431 TI - Identification and distribution of aspartoacylase in the postnatal rat brain. AB - Aspartoacylase metabolizes N-acetylaspartic acid to produce L-aspartate and acetate. An aspartoacylase deficiency in humans is responsible for Canavan disease, a lethal autosomal recessive leukodystrophy. The role of aspartoacylase in the mammalian brain is unclear. Here we have generated and characterized a highly specific polyclonal antibody against aspartoacylase which recognizes a 37 kDa monomer and a dimer in normal but not in aspartoacylase-deficient rat tissue. Aspartoacylase protein expression sharply increases at P14, peaks at P28 and plateaus thereafter. Biochemical analysis reveals immunoreactivity in cytosolic but not in membrane fractions. Histologically, most abundant expression was observed in white matter tracts and thalamus. On the cellular level, aspartoacylase immunoreactivity is restricted to oligodendrocyte somata in both white and gray matter. PMID- 14534432 TI - Visual processing of facial affect. AB - To evaluate the role of the fusiform gyrus in identifying and processing facial emotional expression in humans, MEG data were collected while six healthy subjects judged whether photographs of faces displayed emotion (happiness or disgust) compared to neutral faces and equiluminant scrambled faces. For all six subjects, a magnetic source localizing to right fusiform gyrus was evident approximately 150 ms following presentation of face stimuli, but not following non-face stimuli. MEG source strength for this component was greatest for happy, intermediate for disgust, and lowest for neutral facial expressions, suggesting that activity in fusiform gyrus is sensitive to both face-specific stimuli and to the affective content of the face. These findings are considered in the context of a specialized neural face-dependent information system. PMID- 14534433 TI - Neural expression and regulation of NSSR1 proteins. AB - Neural salient serine-/arginine-rich protein 1 (NSSR1) is a newly identified SR protein that regulates pre-mRNA splicing. In the present study, we demonstrated the neural specialization of NSSR1 protein expression in humans and mice. Strong immunoreactive signals to NSSR1 were observed in mouse cerebral neurons, cerebellar Purkinje cells, pyramidal neurons in CA1, CA2 and CA3 regions of the hippocampus and granule cells in the dentate gyrus. In primarily cultured mouse neural progenitor cells (NPCs), at the undifferentiated status, NSSR1 transcripts were detected, but not the proteins. In comparison, in differentiated NPCs both NSSR1 transcripts and proteins were expressed and significantly up-regulated. The results suggest that NSSR1 is important in regulation of brain function and neural differentiation, possibly via regulating the neural-specific alternative splicing of genes. PMID- 14534434 TI - Novel distribution of calcitonin gene-related peptide in rodent subcommissural organs. AB - For the first time we showed the presence of CGRP immunoreactivity in the SCO of golden hamsters, Wistar rats and Mongolian gerbils. In hamsters, the intense CGRP like immunoreactivity was detected in the whole SCO, including Reissner's fiber- and granule-like structures at the ventricular surface of the SCO. The CGRP positive hypendymal cells were frequently in contact with local blood vessels and some of them were situated intimately to the leptomeningeal space. In the SCOs of rats and gerbils, only the supranuclear area and apical cytoplasm of the ependymal cells were positive for CGRP, whereas the basal pole of the cells and the hypendyma were CGRP-negative. The existence of CGRP in rodents SCO was confirmed by the expression of CGRP mRNA in rat SCO by RT-PCR. The present results indicate that the SCOs of rodent species contain CGRP that may be in part synthesized by ependymocytes themselves. A species difference in the CGRP distributions of rodents SCOs may therefore imply its different synthesis rates or release pathways. PMID- 14534435 TI - Responses to noxious stimuli in mice lacking alpha(1d)-adrenergic receptors. AB - Nociceptive behaviors were examined in the mice lacking alpha1d-adrenergic receptor (alpha1d-AR) and wild type littermates using tail-flick, hot-plate (hindpaw-licking and jumping), tail-pinch and formalin tests. The distribution of alpha1d-AR was studied using in situ hybridization in the wild type mice. Mutant mice showed longer tail-flick and hindpaw-licking latencies while their jumping latency was shorter. Mechanical and chemical nociception was not altered in alpha1d-knockout mice. In situ hybridization study revealed dense alpha1d-AR mRNA expression in the reticular thalamic nucleus, the hippocampus, the cingulate cortex and the spinal cord. These results suggest that alpha1d-AR in the spinal cord contributes to thermal pronociception; and that the jump behavior seen when escaping from heat is inhibited via the supraspinal alpha1d-AR. PMID- 14534436 TI - The Ro52/SS-A autoantigen has elevated expression at the brain microvasculature. AB - An expression cloning technique was used to identify proteins selectively expressed at the blood-brain barrier that may mediate interactions between the brain microvasculature and IgG molecules. Ro52/SS-A is an IgG-binding protein and is an autoantigen implicated in Sjogren's syndrome and systemic lupus erythematosus. The present studies describe the expression cloning of this gene product from a bovine brain microvascular cDNA library. The Ro52 protein interacted with IgG molecules independent of specificity suggesting a potential role as a general antibody receptor. Northern blotting shows the Ro52/SS-A transcript is enriched in the microvascular compartment of brain. The selective expression of Ro52/SS-A in brain at the microvasculature may play a role in brain vascular involvement in autoimmune diseases. PMID- 14534437 TI - Increased serum and cerebrospinal fluid FGF-2 levels in amyotrophic lateral sclerosis. AB - Lack of growth factors and hypoxia are two recent hypotheses about mechanisms underlying motor neuron death in amyotrophic lateral sclerosis. With this background, serum from 15 patients with amyotrophic lateral sclerosis and 15 controls, and CSF from 15 patients and 10 controls were analysed for fibroblast growth factor 2 (FGF-2) using an immunoassay. Serum FGF-2 levels were higher in the patient group than in the control group. FGF-2 was detected in CSF in 11/15 patients, but in none of the 10 control subjects. There were no correlations between age, duration of disease or clinical rating and FGF-2 levels. The findings indicate that FGF-2 is involved in the pathophysiological chain of events in this disorder. PMID- 14534438 TI - Auditory magnetic responses of healthy newborns. AB - We recorded magnetic brain activity from healthy human newborns when they heard frequency changes in an otherwise repetitive sound stream. We were able to record the magnetic counterpart of the mismatch negativity (MMN) previously described only with electric recordings in infants. The results show that these recordings are possible, although still challenging due to the small head size and head movements. The modelling of the neural sources underlying the recorded responses suggests cortical sources in the temporal lobes. PMID- 14534439 TI - Boosting GABA improves impaired auditory temporal resolution in the gerbil. AB - Elderly humans often not only experience peripheral hearing loss but also suffer from more central deficits in temporal auditory processing affecting speech perception. Impaired auditory temporal resolution has also been observed in old rodents. Other studies have demonstrated a reduction of GABAergic function in the auditory pathway of old animals. Here we test the hypothesis that deficits in the GABAergic system affect central auditory processing. Our data suggests that pharmacological augmentation of the GABAergic system ameliorates impaired temporal auditory processing in the gerbil and might be a strategy for the treatment of at least some forms of central hearing loss in humans. PMID- 14534440 TI - Free radical scavenger protects against inner hair cell loss after cochlear ischemia. AB - We investigated the protective effects of edaravone, a free radical scavenger, against ischemic damage of inner hair cells (IHCs) in gerbils. Cochlear ischemia was induced in the animals by occluding the vertebral arteries bilaterally for 15 min. Edaravone (1 mg/kg, i.v.) or saline was administered 1 h after ischemia. Hearing was assessed by auditory brain response (ABR). In animals treated with saline, the ABR threshold shift was 24.1 dB and there was a 26.5% decrease in the number of IHCs. By contrast, in animals treated with edaravone, the threshold shift was 7.5 dB and only 8.8% of IHCs was lost. These results suggest that edaravone protects against damage to the inner ear following transient ischemia. PMID- 14534441 TI - Noise induced hearing loss. PMID- 14534442 TI - Fibromyalgia and the conundrum of disability determination. PMID- 14534443 TI - Immunological findings in a group of coke-oven workers exposed to polycyclic aromatic hydrocarbons. AB - Coke-oven workers are exposed to a high concentration of polycyclic aromatic hydrocarbons, which may change the immunologic responses. In this study, we evaluated humoral immunity by measuring serum IgA, IgE, IgG, and tumor necrosis factor-alpha in 251 coke-oven workers and 89 rolling steel workers in Taiwan. Mean values of serum IgE and tumor necrosis factor-alpha levels were 178.8 IU/mL and 8.4 pg/mL in coke-oven workers, respectively, which were significantly higher than in rolling steel workers (102.6 IU/mL and 2.0 pg/mL; P=0.003 and <0.001). In contrast, serum IgA levels were significantly lower in coke-oven workers than in rolling steel workers (mean=264.7 vs 312.0 mg/dL, P<0.001). Our findings suggest that polycyclic aromatic hydrocarbons exposure may alter the immune responses in coke-oven workers. PMID- 14534444 TI - Unexplained symptoms after terrorism and war: an expert consensus statement. AB - Twelve years of concern regarding a possible "Gulf War syndrome" has now given way to societal concerns of a "World Trade Center syndrome" and efforts to prevent unexplained symptoms following the most recent war in Iraq. These events serve to remind us that unexplained symptoms frequently occur after war and are likely after terrorist attacks. An important social priority is to recognize, define, prevent, and care for individuals with unexplained symptoms after war and related events (eg, terrorism, natural or industrial disasters). An international, multidisciplinary, and multiinstitutional consensus project was completed to summarize current knowledge on unexplained symptoms after terrorism and war. PMID- 14534445 TI - Association between wellness score from a health risk appraisal and prospective medical claims costs. AB - This study examines how wellness scores generated from the Health Risk Appraisal are associated with prospective medical claims costs, controlling for age, gender, and disease status. The study was conducted among 19,861 active employees who participated in the Health Risk Appraisal and selected indemnity or PPO medical plans from 1996 to 1998. A multiple regression model based on group averages of age, gender, disease status, and wellness score levels was developed among a randomly selected screening subsample (n=10,172) from the study sample. Total medical claim costs of -$56, $88, and $3574 were estimated for one additional point on the wellness score, 1 year of additional age, and an existing major disease, respectively. No significant differences were found between the model predicted and actual medical claims costs for the individuals in both screening and calibration (n=9689) subsamples. PMID- 14534449 TI - Effects of firm size on risks and reporting of elevation fall injury in construction trades. AB - Although many occupational safety programs are targeted toward large firms, the construction industry is dominated by smaller firms. This study examines the differential effect of firm size on the risk and the reporting of over 3000 serious and minor nonfatal elevation fall injuries in Danish construction industry trades (1993 to 1999). Small firms (<20 employees) accounted for 93% of all firms and 55% of worker-years. There was an inverse relationship between firm size and serious injury rates and a direct relationship between firm size and minor injury rates. An inverse relationship between firm size and injury severity odds ratios (serious versus minor) was found for carpentry, electrical work, general contracting, and the remaining other trades. Health and safety issues, legislation, and enforcement in the construction industry should, to a greater degree, be focused on smaller firms. PMID- 14534447 TI - Cancer incidence among union carpenters in New Jersey. AB - A cohort of 13,354 male union carpenters in New Jersey was linked to cancer registry data to investigate cancer incidence during 1979 through 2000. Surveillance, Epidemiology and End Results data were used to calculate standardized incidence ratios (SIRs). A total of 592 incident cancers were observed among this cohort (SIR=1.07), which was not statistically in excess. However, significant excesses were observed for cancers of the digestive system and peritoneum (SIR=1.24) and the respiratory system (SIR=1.52). Workers in the union more than 30 years were at significant risk for cancers of the digestive organs and peritoneum (SIR=3.98), rectum (SIR=4.85), trachea, bronchus, and lung (SIR=4.56), and other parts of the respiratory system (SIR=11.00). Testicular cancer was significantly in excess (SIR=2.48) in analyses that lagged results 15 years from initial union membership. Additional etiologic research is needed to evaluate possible occupational and nonoccupational risk factors for testicular cancer. PMID- 14534448 TI - Lung function in Pakistani welding workers. AB - Our objective was to study the effects of welding fumes and their duration of exposure on lung function. This was a matched case-control cross-sectional study of spirometry in 50 nonsmoking manual metal arc welders who work without the benefit of welding fume control ventilation or respiratory protective devices. Welding workers, with exposures longer than 9 years, showed a significant reduction in spirometry (forced expiratory volume in one second [FEV1], FEV1/forced vital capacity [FVC%], and peak expiratory flow [PEF]) relative to controls. Lung function in nonsmoking welding workers is impaired and stratification of results shows a dose-effect of years of welding on lung function. This effect primarily shows an obstructive pattern of airways disease. Preventive measures in developing countries have to be adopted to avert long-term lung damage in welders. PMID- 14534450 TI - Safety practices, neurological symptoms, and pesticide poisoning. AB - Depression, anxiety, inability to concentrate, and spatial disorientation associated with pesticide poisoning may influence farmers' ability to comply with established safety procedures. The purpose of this article is to describe the relationship between safety practices, neurological symptoms, and pesticide poisoning. A survey of farm residents was conducted in an eight-county area in Colorado. Multivariate logistic regression models were used to determine associations between safety practices, neurological symptoms, and previous pesticide poisoning. A number of safety practices were associated with the following neurological symptoms: difficulty concentrating; feeling irritable; relatives noticing memory difficulties; and difficulty understanding reading materials. The associations between safety practices and neurological symptoms were increased in the presence of pesticide poisoning. Factors associated with failure to engage in established safety practices in this study were neurological symptoms. PMID- 14534451 TI - Malingered neurocognitive dysfunction in neurotoxic exposure: an application of the Slick criteria. AB - Toxic torts are increasing across the country and often the results of the neuropsychological evaluation are crucial for defining damages. Therefore, the accurate differentiation of those damaged by toxic exposure from those exaggerating or fabricating deficits is important. However, there is little research on malingering in this context. Presented are four patients claiming cognitive deficits after apparent occupational neurotoxic exposure who were diagnosed as malingering using the Slick, Sherman, and Iverson criteria. The goals of this article were to (1) illustrate the application of the Slick Criteria; (2) discuss current knowledge about the neurological and neurocognitive effects of toxic substances and its impact on clinical decision-making; (3) discuss the application of the Slick Criteria, specifically, and malingering research, generally, to toxic exposure cases; and (4) propose a paradigm in which medical, toxicological and neuropsychology professionals coordinately evaluate cases of alleged neurotoxic chemical exposure. PMID- 14534452 TI - Risk of laryngeal cancer by occupational chemical exposure in Turkey. AB - Laryngeal cancer is the second most common cancer among men in Turkey. In this hospital based case-control study, we evaluated laryngeal cancer risks from occupational chemical exposures. We analyzed 940 laryngeal cancer cases and 1519 controls. Occupational history, tobacco, and alcohol use and demographic information were obtained by a questionnaire. The job and industries were classified by special seven-digit codes. We calculated odds ratios (ORs) and 95% confidence intervals (CIs) based on a developed exposure matrix for chemicals, including diesel exhaust, gasoline exhaust, polycyclic aromatic hydrocarbons (PAHs), formaldehyde, and solvents. An excess of laryngeal cancer occurred with diesel exhaust (OR=1.5, 95% CI=1.3-1.9), gasoline exhaust (OR=1.6, 95% CI=1.3 2.0), and PAHs (OR=1.3, 95% CI=1.1-1.6). There was a dose-response relationship for these substances with supraglottic cancers (P<0.000). The PAH association only occurred among those who also had exposure to diesel exhaust. PMID- 14534453 TI - Estimating the induction period of pleural mesothelioma from aggregate data on asbestos consumption. AB - This study aimed to estimate the induction period from causal action of asbestos exposure to the manifestation of mesothelioma. We included the 9 countries for which we could find published aggregate data on the use of raw asbestos for a relevant time period. We extracted the annual numbers of cases of pleural cancer among men from the World Health Organization mortality database for those years using the International Classification of Diseases, 9th revision, classification. For the Scandinavian countries, we used published national cancer incidence data. In autoregressive Poisson regression modeling, we invoked different time lags of the mean annual use of asbestos to specify which time span produced the best correlation between the 2 time series. The ecologic analysis suggested that the most probable estimate for the mean induction period (use versus morbidity at society level) is approximately 25 years. PMID- 14534455 TI - The gap between actual practice and evidence-based "best practice". PMID- 14534454 TI - Primary congenital hypothyroidism, newborn thyroid function, and environmental perchlorate exposure among residents of a Southern California community. AB - The objectives of this study were to evaluate whether there were higher rates of primary congenital hypothyroidism (PCH) or elevated concentrations of thyroid stimulating hormone (TSH) in a community where perchlorate was detected in groundwater wells. The adjusted PCH prevalence ratio and 95% confidence interval (CI) comparing the study community to San Bernardino and Riverside counties combined was 0.45 (95% CI=0.06-1.64). The odds ratios for elevated TSH concentration were 1.24 (95% CI=0.89-1.68) among all newborns screened and 0.69 (95% CI=0.27-1.45) for newborns whose age at screening was 18 hours or greater. Age of the newborn at time of screening was the most important predictor of the TSH level. These findings suggest that residence in a community with potential perchlorate exposure has not impacted PCH rates or newborn thyroid function. PMID- 14534456 TI - Ten ways to retain per diem staff. AB - Recruiting and retaining part-time staff require unique considerations. This article reflects my suggestions based on recent experiences as a part-time home care staff nurse and my previous role as a home care manager. PMID- 14534457 TI - ACHC, CHAP, or the joint commission: which one is for you? AB - Selecting the right accreditation organization is an important decision. This month's column reviews the three nationally recognized accreditation organizations, and examines their standards and survey activities. Factors home care organizations should consider when choosing an accreditation body are presented to facilitate knowledgeable decision making. PMID- 14534458 TI - Making nutrition sense from OASIS. AB - Assessing nutrition is an integral part of the comprehensive assessment for each patient. Although OASIS does not have a specific section or identified items labeled for nutrition, there are numerous nuances of nutritional status imbedded in the OASIS-B1. This article will identify the many OASIS items that contribute to the nutritional assessment and point out when OASIS responses should trigger a referral to a registered dietitian. PMID- 14534459 TI - Magnet home care agencies: a professional way to impact quality and retention. AB - By 2009 the nursing shortage will reach a critical level with home care agencies seriously affected. Hospitals have begun to embrace one proven method for recruiting and retaining nurses: achieving magnet recognition from the American Nurses Credentialing Center. Although magnet recognition has been available to home health agencies since 2000, no agency has applied. This article outlines how agencies can take advantage of the program. PMID- 14534460 TI - Cultural assessment and care planning. AB - Culturally competent care adapts care to the patient's cultural needs and preferences and begins with a cultural assessment that forms the care plan's foundation. Nurses who assess their patients' cultural beliefs, values, and practices are better able to individualize care and achieve positive outcomes. This article describes a cultural assessment tool, strategies for obtaining cultural assessment data, and a process for creating a culturally appropriate care plan. PMID- 14534461 TI - Using the new OASIS-based reports in OBQI. AB - OASIS-derived outcome reports first became available to home care agencies in February 2002 and depicted the agency's current (observed) outcome achievement rate as compared to a reference (predicted) rate on 41 separate outcomes. New three-bar reports are now available that allow agencies to evaluate their current outcome rates against their previous year's rates and a reference rate. This article compares the newly-available three-bar reports with the prior two-bar reports, including information provided in those reports in the OBQI process. PMID- 14534462 TI - New tool to use with patient tally reports. AB - A new tool is available for agencies to use with their patient tally reports derived from OASIS data. By using queries submitted to the outcome and case mix tally reports, agencies can select a group of care episodes for follow-up in the process-of-care investigation portion of their outcome enhancement activities. An example of how an agency used the tool to select care episodes for review is provided. PMID- 14534463 TI - Learning to have difficult conversations leads to increased hospice referrals. PMID- 14534464 TI - Depression is more than just an OASIS item. PMID- 14534465 TI - [The silent revolution of the elderly. Interview by Veronique Girault]. PMID- 14534466 TI - [Colorectal cancer: assessment of two-yearly screening of the population in health care examination centres]. AB - OBJECTIVE: To present the first assessment of the two-yearly faecal occult blood test used to screen patients attending health care centres for examinations. Method The Hemoccult II test was proposed by correspondence to 26 530 subjects aged 52-74 years, having exhibited a negative test during their periodical check up 2 years before. RESULTS: The test was performed by 20 590 subjects and the corrected participation rate, taking into account unjustified sending, was of 83.1%. The positivity rate was 3.5%. In the 519 colonoscopies performed (71% of positive Hemoccult II), 23 cancers and 91 adenomas (31 larger than 1 cm) were observed (positive predictive value=22%). In 213 subjects, the information on the follow-up of the positive Hemoccult II were unavailable or unexploitable (lost to follow-up, coloscopy refused, inappropriate examinations). DISCUSSION: These results show that the two-yearly screening for colorectal cancer in an informed population after a periodical check-up results in a good participation rate. The follow-up rate of positive subjects, fundamental for the assessment of the program, is lower than during the regular check-up but remains relatively satisfactory. PMID- 14534467 TI - [Ogilvie syndrome in the elderly: retrospective study of 40 cases]. AB - OBJECTIVE: To assess, in a population of elderly patients, the circumstances clinical and progressive characteristics and form of management of an acute colonic pseudo-obstruction (ACPO). METHOD: Retrospective study of the files of 40 patients aged 70 and more having presented an acute colonic pseudo-obstruction and hospitalised in the university hospital in Dijon from January 1995 to June 2000. RESULTS: The population was composed of 24 men (60%) and 16 women (40%) with a mean age of 80.8 years. The reasons for hospitalisation were varied: only 15 patients had been referred for an occlusive syndrome. 39 patients presented with abdominal distension, 30 suffered from abdominal pain. In this population, 17 patients exhibited reduced or clearly limited mobility, 20 patients had altered cognitive capacity with an MMS < 20. Thirty-eight patients suffered from a progressive heart disease and 8 patients presented with advanced dementia. The usual treatment of these patients to help the intestinal transit included diuretics in 25 cases and agents slowing the transit in 19 cases. Biologically speaking, hypokalaemia was observed in 21 cases and increased thyroid stimulating hormone (TSH) in 3 cases. An image was obtained of the abdomen without preparation in all patients: the mean caecum diameter was of 9.8 cm. A colon aspiration was performed in 20 patients and molecules to improve peristaltism were administered in 21 cases, with neostigmine prescribed for 9 patients. Surgery was required for 7 patients and 3 of them subsequently died. In terms of progression, 20 patients were cured after treatment, 13 relapsed and 7 patients worsened. Unfortunately, 10 patients died in our series. DISCUSSION: In our study, the clinical profiles of acute colonic pseudo-obstruction were similar to those described in the literature: they occurred in varying circumstances, in medical or surgical settings ina predominantly male population of elderly, heavily dependent,patients. Treatment of this affection is not clearly codified for fragile patients with multiple diseases and the mortality rate observed should stimulate further studies, notably on pharmacological level. PMID- 14534469 TI - [Are Egyptians pioneers of auscultation?]. PMID- 14534468 TI - [Upper mesenteric venous thrombosis revealing familial resistance to activated protein C]. AB - INTRODUCTION: Mesenteric venous thrombosis is a rare disease but potentially severe because of the prognosis of intestinal infarction with high mortality rate (60%). OBSERVATION: We report the case of a 55 year-old man who presented with an upper mesenteric venous thrombosis related to a familial resistance to C reactive protein through factor V Leiden mutation. COMMENTS: The discovery of a mesenteric venous thrombosis requires aetiological research that is usually multifactorial. Among the most frequent genetic coagulation abnormalities observed resistance to C reactive protein due to G202110A prothrombin gene mutation is the most common. Although factor V Leiden mutation is less frequent, it requires anticoagulation therapy for life in the case of the appearance of a thrombosis. PMID- 14534470 TI - [Specific immunotherapy for respiratory allergies. A modern treatment]. AB - AN EFFICIENT AETIOLOGICAL TREATMENT: Specific immunotherapy or allergen vaccination currently constitutes the only aetiological treatment for allergic respiratory diseases and is more effective in children than in adults. Since its introduction in 1911, it has been much disparaged in spite of its effectiveness demonstrated in double blind placebo controlled studies, for the treatment of both allergic rhinoconjunctivitis and stable and controlled asthma. MECHANISM OF ACTION: Its mechanism of action varies depending on the type of allergen (venoms or pneumallergenes) and its administration route (subcutaneous or local) and recent studies indicate that the prevalent mechanism proceeds from a modification of the T cell response by an immunological deviation (stimulation of the Th0/Th1 lymphocytes) or T lymphocyte anergy (reduction in Th0/Th2) or both at same time. The IL-10, immunosuppressive cytokine secreted by the third population of T-Tr1 lymphocytes, would have an important role. IN PRACTICE: The indications for specific immunotherapy have been specified in international consensuses, and its practical administration must respect preconditions and a certain number of rules. Improvement in quality of allergenic extracts by standardization, better knowledge of its mechanism of action and the availability of local administration routes reinforce its place in the global therapeutic strategy for the management of respiratory allergies. A greater use of the sublingual route, together with biological and clinical research will allow the widening of its indications. PMID- 14534471 TI - [Therapeutic use of anti-TNF-alpha agents in spondyloarthropathies]. AB - CLASSICAL DATA: Spondyloarthropathies regroup several rheumatological entities (ankylosing spondylitis, reactive arthritis, psoriatic rheumatism, entero colopathic disease rheumatism, undifferentiated spondyloarthropathies ) with validated diagnosis criteria. Drug therapy is based upon NSAIDs (non-steroidal antiinflammatories). Refractory forms may lead to severe functional impairment, raising the need of more effective treatments. IN FAVOUR OF ANTI-TNF-ALPHA AGENTS: Several arguments (TNF-alpha serum levels, elevated levels of mRNA, TNF messengers, in sacro iliac biopsies, efficacy of anti TNF-alpha agents in Crohn's disease ) justify the use of anti-TNF-alpha agents in the treatment of spondyloarthropathies. Two biologic agents have been assessed in these circumstances: a monoclonal antibody (Infliximab) and a soluble form of the TNF receptor (Etanercept). EFFICACY AND SAFETY: Results of open and controlled studies, although on small series, demonstrated the significant efficacy of anti TNF-alpha agents on the various clinical, biological, functional and quality of life parameters, and confirmed by imaging (MRI ). Tolerance is fair, but two cases of diffuse tuberculosis have been reported with Infliximab. THERAPEUTIC PROGRESS: Even if additional studies are required to answer some questions (long term efficacy and safety, treatment modalities), anti TNF agents appear as a therapeutic progress in refractory spondyloarthropathies, for which few validated options have existed up till now. PMID- 14534472 TI - [Admission to intensive care of very old patients]. AB - THE EXTENT OF THE PROBLEM: Because of the costs of intensive care, there is a possibility that access to this sector may be limited for those above a certain age. It would therefore appear necessary to develop an ethical clinical strategy in order to assess as precisely as possible and on individual level, the benefits and risks of these techniques, since age itself is simply a criterion among so many others. Among the elements that are important for the decision is the respect of the patient's wish, when it can be obtained since this is a fundamental point. In studies conducted in the United States, 50 to 90% of the elderly persons interviewed did not wish to be resuscitated in the case of cardiac arrest. CRITERIA FOR ADMISSION: The decision to admit an elderly patient in an intensive care unit must take into account the functional state of the patient, appreciated on daily activity and mobility scores and the neuro psychological assessment, before hospitalization. In parallel, the severity of the underlying disease and the impact on visceral failures, assessed by the severity scores on admission appear to be more reliable prognostic elements than the patients' age itself. FOLLOWING RESUSCITATION: The quality of life of elderly patients within the months following resuscitation is difficult to assess, but is considered as acceptable in the majority of surviving patients. IN PRACTICE: The choice of admission in intensive care of an elderly patient requires a multidisciplinary approach that takes into account the patient's and/or family's wishes, the benefit/risk ratio of the technical act but which also, in certain cases, bears in mind the principle of end of life and the patient's dignity. PMID- 14534473 TI - [New strategies in the care of sore throat with fast diagnosis tests and short therapies]. PMID- 14534474 TI - [Acute mastoiditis in children: clinical and bacteriological study of 17 cases]. AB - OBJECTIVE: Acute mastoiditis (AM) represents the most frequent complication of acute otitis media (AOM) in children. In the literature, its incidence is stable but with an impressive increase in pneumococci with reduced sensitivity to penicillin (PRSP). The aim of this study was to assess the incidence of AM in the area of Tours and the prevalence of PRSP. METHOD: This was a retrospective study of children admitted to the regional paediatric centre in the Tours area, between January 1994 and May 2001. The coded AM files were analysed, excluding all the sub-acute forms or those complicating a cholesteatoma. The criteria studied concerned the clinical signs on admission, the imaging data and the bacteriological samples and the results of treatment. RESULTS: Seventeen children (8 boys, 9 girls) were admitted for an AM during the study period. Their mean age was of 3.2 years (range: 6 months to 13 years). In 24% of cases, the mastoiditis existed on admission and in 59% of cases it complicated an AMO already treated with antibiotics. In 3 cases (18%), the AM was complicated on admission with peripheral facial paralysis in one case and thrombosis of the lateral sinus in 2 cases. The germ responsible was identified in 14 cases (82%) with a predominance of pneumococci (11 cases). Eight were PRSP-type. Scan of the pars petrosa identified a subperiosteal abscess in 13 cases. Fourteen mastoidectomies were performed and the mean duration of antibiotic therapy was of 23 days. The outcome was always good. CONCLUSION: Over the past 7 years, the incidence of AM has been estimated at 1.2/100 000 children aged under 15 per year. The prevalence of PRSP is high but does not change the principles or the results of the treatment of AM. PMID- 14534475 TI - [The general practitioners' emergency bag; what is needed for life-threatening emergencies?]. AB - OBJECTIVE: The emergency bag is an essential tool for the general practitioner and its contents vary depending on the practitioners' activity. The objective was to compare the emergency bags with those that are recommended in the literature and to define what is necessary in the case of life-threatening emergencies. METHOD: A survey on the general practitioners' emergency bag was conducted in the Cote d'Or area during the month of September 2000. RESULTS: Globally, emergency bags were not always adapted to the practitioners' activity and expiry dates were not systematically checked. CONCLUSION: Three types of emergency bags were defined depending on three areas of activity: urban, semi-rural and rural. PMID- 14534476 TI - [Primary hypertrophic osteoarthropathy without cutaneous involvement (Currarino's disease)]. AB - INTRODUCTION: Hypertrophic osteoarthropathy (HOA) is a syndrome associating hippocratic fingers, arthropathy and periostosis of long bones. Currarino's disease, considered at present as a clinical form of primary HOA, is characterized by the absence of pachydermia. OBSERVATION: A 24-year-old Caucasian man, consulted for a painful swelling of both ankles that had developed over the past year. Clinical examination revealed hippocratic fingers without pachydermia. The ankles were swollen. The X-rays showed periosteal apposition and an acro osteolysis. In view of this triad: arthropathy, hippocratic fingers and periostosis, primary HOA without cutaneous involvement or Currarino's disease was diagnosed. The search for a secondary cause remained negative. Clinical improvement was obtained after 15 months with non-steroidal anti-inflammatory drugs (NSAID) and colchicine. COMMENTS: Although exceptional, primary HOA without cutaneous involvement is a genetic disease which must not be ignored. PMID- 14534477 TI - [Bilateral blindness and sphenoethmoid sinusitis]. PMID- 14534478 TI - [Monoarthritis with Haemophilus parainfluenzae septicaemia during multiple myeloma of the bones]. PMID- 14534479 TI - [Tolosa-Hunt syndrome]. PMID- 14534480 TI - [Portal vein thrombosis]. AB - DIAGNOSTIC CIRCUMSTANCES: Portal vein thrombosis is the second cause of portal hypertension after cirrhosis in Western countries. Diagnosis can be either made at the acute stage in the context of abdominal pain or after appearance of a porto-portal collateral venous circulation leading to the formation of a portal cavernoma, the diagnosis being made in the circumstance of rupture of oesophageal varicose veins or manifestations of hypersplenism. AETIOLOGICAL SURVEY: In the absence of hepatocellular carcinoma, causes that need to be investigated are cirrhosis, local factors (intra-abdominal sepsis, abdominal surgery, splenectomy or pancreatitis), and one or several prothrombotic affections (acquired or inherited prothrombotic states are present in 70% of cases, with myeloproliferative disease ranking first). REGARDING TREATMENT: Anticoagulant therapy generally allows recanalisation of the thombosed veins in recently constituted thrombosis. Some patients at the portal cavernoma stage can also benefit from anticoagulant therapy: patients with a prothrombotic state without large oesophageal-gastric varicose veins. In the case of large oesophageal gastric varicose veins that have never bled, treatment to prevent haemorrhages due to portal hypertension according to the same modalities as in cirrhosis must be associated with the prescription of an anticoagulant. In the absence of prothrombotic affection or in patients having already suffered from haemorrhages due to portal hypertension, the benefit of anticoagulant therapy is less clearly established. PMID- 14534481 TI - [The psychiatric forms of Creutzfeldt-Jakob disease]. AB - CLINICAL FEATURES: Frequent in the new variant of Creutzfeldt-Jakob disease, psychiatric disorders are less common in the sporadic form. The disorders are non specific: mood disorders, personality changes and psychotic symptoms. DIAGNOSIS: The diagnosis of Creutzfeldt-Jakob disease is evoked on the emergence of the first neurological signs, notably ataxia and myoclonia. The diagnosis is supported by the search for P 14-3-3, EEG in the cerebrospinal liquid as well as electroencephalogram and magnetic imaging that permit distinction between the sporadic and the new variant forms. PMID- 14534482 TI - [Management of sexual dysfunction in patients with coronary heart disease]. AB - THE CONTEXT: Patients with coronary heart disease are generally males aged more than 55 and in whom the question of sexual activity must be evoked, not only with regard to the risks involved with the sexual act itself but also regarding the management of an eventual erectile dysfunction. POSITIVE DATA: A negative and maximal for the age stress test is of predictive value and can eliminate the hypothesis of recurrent coronary ischaemia during sexual intercourse. Drug induced effects on the libido and erectile function are known but only led to suspension of treatment in one out 438 patients treated for one year. THERAPEUTIC MANAGEMENT: In the case of documented erectile dysfunction, the combination of sildenafil nitrate derivatives or NO suppliers is formally contra-indicated because of the risk of hypotension. Post-marketing registers and specific studies in patients with coronary heart disease demonstrate the good haemodynamic and coronary tolerance to sildenafil in this category of patient, so long as the contra-indications are respected. The Princeton Consensus Panel has proposed a therapeutic strategy adapted to each patient and according to their level of risk and its treatment. PMID- 14534483 TI - [Skin ageing and its prevention]. AB - INTRINSIC AND EXTRINSIC FACTORS: Skin ageing is due to the conjunction of intrinsic (chronological ageing) and extrinsic factors (fundamentally photo ageing). The physiopathological mechanisms of intrinsic ageing rejoin those of the ageing of all the other organs. Among the intrinsic causes, tobacco and above all ultra-violet radiation, UVB and also UVA, play a preponderant role. Photo ageing is secondary to complex mechanisms that are increasingly known. The UVB directly interact with the DNA of the cutaneous cells. The deleterious effects of UVA are principally due to the formation of free radical oxygen, which result in an alteration in the nuclear and also mitochondrial DNA, but also an activation of the enzymes, metalloproteinase, capable of damaging the extra-cellular matrix. DELETERIOUS CONSEQUENCES: The phenomena of ageing provoke the decline in defence, healing and perception mechanisms and in the thermoregulation of the skin tissue. There are numerous and often unsightly clinical manifestations. Photo-ageing can be considered as a marker of risk of photo-carcinogenesis requiring increased clinical surveillance. PREVENTIVE AND CURATIVE MEASURES: The prevention of skin ageing must be based on the use of sunscreens protecting against both UVB and UVA, but, in order for them to be effective, they require a change in general life style. There are many efficient therapeutic means, but the possible side effects must be known and explained to the patient. Retinoids, in view of their innocuousness and efficacy not only in prevention but also treatment of skin ageing, should be considered as a therapeutic option of choice. PMID- 14534484 TI - [Hepcidin, the conductor of iron homeostasis]. PMID- 14534485 TI - [The interest and assessment of the application of antibiotic protocols in a department of pneumology]. AB - OBJECTIVE: To assess whether antibiotic prescriptions in a department of pneumology are in accordance with the hospital conventions and whether documented prescriptions and probabilistic prescriptions are correctly adapted with regard to microbiological results and clinical progression of patients. METHOD: A 3 month prospective observational study was performed from november 1999 to january 2000 in a department of pneumology on all antibiotic prescriptions. The referential used for the conformity of probabilistic was the guide to pneumology antibiotic protocols regarding the most frequently encountered diseases. Treatment was considered adapted if the choice of the molecule and the dose administered were effective. Documented antibiotherapy was considered appropriate if the dose was correct and/or the micro-organisms were sensitive to at least one of the molecules administered. Probabilistic treatment was considered appropriated if progression was good without any change in the antibiotics. RESULTS: Among the 404 patients hospitalised in Pneumology, 132 (33%) received at least one antibiotherapy. There was a total of 163 treatments, 142 (87%) of which were probabilistic and 21 were documented. Seventy-two percent of the probabilistic prescriptions were in accordance with the protocols. The majority of those which did not conform (60.5%) was due to the dose rather than the choice of the antibiotic. More than three quarters of the probabilistic and documented treatments (79%) were clinically adapted. Treatment failures were more often due to a mistake in spectrum rather than an inappropriate dose. CONCLUSION: The use of antibiotics has become increasingly complex because of the need for therapeutic efficacy, limitation of the selection of microbial resistance, and the cost of treatment. Application of antibiotic protocols drawn-up by the clinicians, microbiologists and chemists concerned appears to be an efficient solution that is clearly acceptable to the various participants. PMID- 14534486 TI - [Chronic obstructive pulmonary disease in France: the patients' perspective. Results of an international survey (Confronting COPD)]. AB - INTRODUCTION: Chronic obstructive pulmonary disease (COPD) has become a real public health problem throughout the world. In spite of its association with high rates of morbidity and mortality, it is not accorded sufficient attention. In this paper, we present the results of the French part of the international survey "Confronting COPD in North America and Europe" concerning the impact of COPD from the perspective patients'. METHODS: Subjects were randomly recruited by means of telephone questionnaires. Patients included in the survey were to be aged > or =45 years with a cumulative cigarette consumption of > or =10 packets-yrs, and having been diagnosed with COPD, emphysema or chronic bronchitis, or whose symptoms fulfilled a definition of chronic bronchitis. RESULTS: According to this survey, the prevalence of COPD among smokers (mean number of packets per year: 38.2 +/- 26.1) was 3.2%. COPD was undiagnosed in 17% of patients in spite of the presence of characteristic symptoms. Respiratory difficulties had a marked adverse effect on the quality of life although patients tended to play down the severity of their problems. DISCUSSION: This survey reveals that COPD appears to be under-diagnosed and under-treated in France, as shown by the results of the same survey in the other 7 countries taking part. Considerable efforts are thus required to improve the speed and precision of diagnosis and to improve management of these patients. PMID- 14534487 TI - [Association of distal tubular acidosis, Hashimoto's thyroiditis and Gougerot Sjogren's syndrome]. AB - INTRODUCTION: Distal tubular acidosis is associated with auto-immune diseases not specific to organ. The coexistence of distal tubular acidosis and auto-immune thyroid affection is very rare. OBSERVATION: A 36-year-old woman exhibiting primary hypothyroidism, Gougerot-Sjogren's syndrome and hypergammaglobulinemia, presented distal tubular acidosis revealed by severe hypokaliemia and complicated by quadriplegia and circulatory arrest. Correcting the thyroid defect did not appear to influence the progression of acidosis. COMMENTS: Based on this observation, one can discuss the pathogenesis of distal tubular acidosis during auto-immune diseases (hypothyroidism, Gougerot-Sjogren's syndrome and monoclonal hypergammaglobulinemia) and its impact on therapy. PMID- 14534488 TI - [Discovery of Chlamydia trachomatis in the throat during diagnosis of secondary syphilis in an HIV+ patient]. AB - INTRODUCTION: Chlamydia trachomatis (CT) pharyngitis has rarely been described in the literature. Studies in the last decade have shown a prevalence of less than 5%, with more women than men infected in the pharynx. Among homosexual men, only one study, conducted more than 5 years ago, detected the presence of CT in the throat (in one patient out of 13 tested). OBSERVATION: A 33 year-old homosexual man consulted for cutaneous lesions on the palms and the chest. Infected by HIV, he was taking antiretroviral therapy but no antibiotic prophylaxis. He admitted practising unprotected (insertive and receptive) fellatio with unknown partners. The lesions were those of secondary syphilis but the location of the chancre remained unknown. An enlarged screening for sexually transmitted infections detected the presence, through molecular amplification, of CT in the throat without further localisation (urethra, anus). After intramuscular injection of Extencillin and an 8-day-regimen of cycline, CT was no longer detected in the throat. The course of HIV infection did not appear modified by this infectious episode. COMMENTS: According to a review of the literature, the 4 most recent studies have not revealed CT in the throat of homosexuals although the bacteria was detected in the urethra of 3 to 4% of them. Chlamydia trachomatis is more frequently detected in the genital tract rather than the pharynx of women also screened in various localisations. These results must be tempered by the participant recruitment methods, diagnostic methods used, sample size and history of recent antibiotherapy. The pharyngeal mucosa might be less receptive to CT than the urethral mucosa. Association of CT pharyngitis with syphilis or its occurrence during HIV infection are not documented. The molecular amplification technique detects CT in the throat with sensitivity and specificity. A one-week regimen of cycline can cure this pharyngitis. Cost-effectiveness of screening for pharyngeal CT has not yet been assessed in persons 'at risk'. PMID- 14534489 TI - [Pleuro-pulmonary metastases of a cerebral glioblastoma]. PMID- 14534490 TI - [Sigmoid neo-vaginal inflammation. Haemorrhagic rectocolitis or diversion colitis?]. PMID- 14534491 TI - [Tetanus vaccination. Re-vaccination is necessary but not always sufficient]. PMID- 14534492 TI - [Epidemiology of atherosclerotic cardiovascular risk in HIV-1 infected patients]. AB - OBVIOUS EFFICACY, BUT ALSO RISKS: The administration of combinations of highly active antiretroviral since 1996 has greatly decreased morbidity and mortality in HIV-1 infected patients. Side effects associated with these treatments were rapidly described. Other than abnormal distribution of fat, the dyslipidemia and insulin-resistance leads to a pro-atherogenic profile in these patients. Since 1998, the publication of several observations of acute cardio-vascular events in HIV-1 infected patients most often treated by antiretrovirals has raised the question of the increased risk of atherosclerosis in these patients. Today, some studies have been published and several are ongoing to evaluate the cardiovascular risk in HIV-1 infected patients. TWO FUNDAMENTAL QUESTIONS: The purpose of this review was to critically report the results of published studies in order to answer the two major questions: do HIV-1 infected patients have an increased cardiovascular risk compared to non infected subjects, and what are these cardiovascular risk factors? REPLY ELEMENTS: The review of available published results is in favour of an increased risk in HIV-1 infected patients compared to the general population. However, it is difficult to evaluate the importance of the absolute risk compared to the benefits of antiretroviral treatments and also the inherent effect of antiretroviral molecules. The main risk factors seem to be classical risk factors such as smoking habits, age and dyslipidemia that are partly due themselves to highly active antiretroviral treatments PMID- 14534493 TI - [Acute higher funghi mushroom poisoning and its treatment]. AB - PRINCIPLE: The various mushroom poisoning syndromes are summarised together with elements underlining uncertainty and lack of knowledge. For each of the classical syndromes concerned, classified in delays inferior or superior to 6 hours, the toxins and their mechanisms of action, the main mushrooms responsible, the symptoms and their treatment are all presented. EARLY SYNDROMES: Characterised by early onset within 6 hours, these represent the majority of intoxications. There are 6 syndromes: gastro-intestinal (resinoid), muscarine (sudorien, cholinergic), pantherine (myco-atropine, anticholinergic), coprine (similar to the antabuse syndrome), narcotine (psilocybin, hallucinatory) and paxillus syndrome (exceptional). LATE SYNDROMES: Characterised by an onset after six hours, they regroup the phalloid syndrome that is responsible for 90 to 95% of deaths due to higher funghi mushrooms, the orellanine and gyromitrin syndrome and new syndromes identified over the past decade concerning acute renal failure with shorter onset than during the orellanine syndrome (Amanita proxima), erythermalgia (Clitocybe amoenolens), rhabdomyolysis (Tricholoma equestre) and central nervous system failure (Hapalopilus rutilans). PMID- 14534494 TI - [The role of the physician when faced with mutilations of the female genitalia]. PMID- 14534495 TI - [Prevention of hepatitis C. How to update the pioneer work done by Eugene Jamot?]. PMID- 14534496 TI - [Are the indications and choice of labile blood products adapted? National survey on prescriber's knowledge]. AB - OBJECTIVE: There is little data available on current practice related to prescription of labile blood products (LBP) by French physicians. The aim of this study was to assess whether prescriptions were conform to Anaes (French Medicine's agency) guidelines, with regard not only to indications but also quality of the products, so as to define the improvements that could be made. METHOD: Thirty-four clinical case reports, classified by specialties were sent to prescribing physicians working in the regional health centers, from 17 different blood banks, from October 1997 to February 1998. The prescribers were requested to answer only the questions that were specific to their particular field of experience. Each case description included multiple choice questions on the indication for transfusion of concentrated of red blood cells (RBC) and/or platelets (CP) and/or plasma, and the possible requirements for specification or modification of the guidelines applicable to these products. The primary end point of analysis was the adequation of the answers to the Anaes recommendations. RESULTS: Answers were obtained regarding 5092 clinical cases from 818 physicians. The participation rate was of 30%. The specialties were as follows: 34% anesthesiologists, 14% oncologists-haematologists, 13% internal medicine specialists, 11% emergency physicians, 10% paediatricians, 8% obstetricians, 7% geriatricians, and 3% transplantation surgeons. Eighty-two percent of the answers came from physicians working in the public health services. The adequation with the indication for transfusion was of 90.3% for RBC, 92.3% for platelets and 93.8% for plasma. The percentages of correct answers regarding the indications for specification or modification of the LBP were as follows: 90.3% were correct for irradiation (of either RBC or platelets); 68.8% and 53.2% respectively for leukocyte depletion from RBC and platelets; 64% for phenotyped RBC; 68.2% for compatibilized RBC; and 57.3% for apheresis platelet concentrates. There was no difference in results depending on the type of center, private or public, and the quality of LBP prescribed. The answers obtained from the anaesthesiologists' clinical cases were less accurate with regard to RBC but more accurate with regard to PC compared with other specialists. CONCLUSION: This study shows the correct management of the indications for transfusion by the prescribing physicians who participated in the study, but the lack of knowledge with regard to the indications for specifications and/or transformations of LBP. The respect of the indications for transfusion is the corner stone of safe transfusion and this phase should be optimized with improved dissemination of information on transfusion and training for the physicians and programs that would improve the quality. PMID- 14534497 TI - [Severe acute respiratory syndrome (SARS). The questions raised by the management of a patient in Besancon and Strasbourg]. AB - INTRODUCTION: In mid-June 2003, 8500 probable cases of Severe Acute respiratory Syndrome (SARS) responsible for 800 deaths in 29 countries were reported by the World Health Organization (WHO). In France, 5 probable cases of SARS were diagnosed until May 2003. We report the case of one of the 5 French cases, hospitalized successively in Besancon then in Strasbourg. OBSERVATION: This 54 year-old man was infected during the Air France flight from Hanoi to Paris via Bangkok by the index case, who himself had been contaminated at the French hospital in Hanoi and who was hospitalized in a critical state in the Tourcoing hospital shortly after his arrival in France. The first clinical signs (fever and dry cough) appeared 4 days after contact with the index case. The chest X-ray showed a pulmonary consolidation 6 days after the first clinical symptoms. Blood test revealed deep lymphopenia and elevated LDH. The patient finally recovered with a treatment combining ribavirin and steroids. CONCLUSION: The clinical symptoms of SARS are unspecific and its evolution may be misleading. The epidemiological context (contact with the index case) is a key point for the diagnosis. The management of such a patient requires the careful protection of contacts, the adequation between medical and paramedical staff and excess work and communication to the general public. PMID- 14534499 TI - [Disseminated cryptococcosis during severe decompensated alcoholic cirrhosis]. PMID- 14534498 TI - [Progressive myelopathy revealing an extra-medullar arteriovenous fistula]. PMID- 14534500 TI - [The management of anemia during cancer. An adapted solution]. PMID- 14534501 TI - [Epidemiological data on anaemia during malignant affections]. AB - FREQUENT IN THE CASE OF CANCER: According to a European multicentre study on 15 000 cancer patients, the percentage of anaemia exceeded 66% during the development of solid tumours and 72% during development of malignant tumours. The lowest levels of haemoglobin were correlated with the lowest performance status, however no treatment had been prescribed for anaemia in the majority of patients. MULTIPLE MYELOMAS AND LYMPHOMAS: The frequency of anaemia appeared just as high during these malignant affections and the correlation between haemoglobin levels and performance status was just as clear. Likewise, around one half of the anaemic patients had not been treated in an adapted or efficient manner. SIGNIFICANCE OF ANEMIA DURING LYMPHOMAS: It appeared that the anaemia of Hodgkin's disease, related to the inflammation, was correlated with shorter survival, whereas the anaemia of non-hodgkin lymphoma, related to medullar infiltration, generally affected the complete rate of response. PMID- 14534502 TI - [The basis of treatment with recombinant human erythropoietin]. AB - EXPERIMENTAL DATA: A line of transgenic mice overexpressing erythropoietin was created. These mice retained their capacity to reduce their gastro-intestinal absorption of iron and to regulate the changes in their iron metabolism and they could serve as a model for the in vivo study of iron homeostasis and erythropoiesis. NEW INDICATIONS FOR RECOMBINANT HUMAN ERYTHROPOIETIN: After chronic terminal kidney failure, the treatment of chronic dialysed kidney failure patients and patients treated with azathioprine or patients having undergone surgery and requiring transfusion, other indications have been proposed. Such as anaemia in children following inadequate production of endogenous erythropoietin and/or direct inhibition of the erythroid cell line in the bone marrow or anaemia during pregnancy and, since the Sixties, anaemia during cancer. TO ASSESS THE PHYSIOPATHOLOGY OF ANEMIA:In anaemic patients suffering from a malignant blood disease, it would be useful to calculate the relationship between the predicted and observed rates of erythropoietin as well as the transferin serum receptors. PMID- 14534503 TI - [The benefits of treatment with recombinant human erythropoietin in cancer patients]. AB - IMPROVED QUALITY OF LIFE WITH EPOETIN BETA: In a study against a placebo, there was evidence that the quality of life scores were significantly improved in patients treated with epoetin beta, whether they exhibited a solid tumour or a malignant lymphoma. The same was noted in children with cancer exhibiting severe neoplasia and treated with chemotherapy. The efficacy and tolerance to treatment were equivalent, whatever the administration regimen. IN PATIENTS SUFFERING FROM MYELODYSPLASTIC SYNDROMES: A particular entity among malignant blood diseases, myelodysplastic syndromes are at the origin of anaemia against which repeated transfusions and growth factors are proposed with varying results and disadvantages, and against which erythropoietin may be moderately effective (a mean of 25% in non-selected cohorts of patients). DEPENDING ON THE PROTOCOLS OF ERYTHROPOIETIN ADMINISTRATION IN CASES OF MYELODYSPLASTIC SYNDROMES: Recombinant human erythropoietin, irrespective of its concentration in myelodysplastic cell culture, does not appear capable of restoring normal erythropoiesis. The influence of prolonged treatment is not admitted by all. The effects of the addition of growth factors (notably G-CSF) are obvious but some are controversial because of the costs and the prolonged duration of such treatments. PMID- 14534504 TI - [The problem of acquired aplastic anaemia]. AB - PURE RED CELL APLASIA: Designated by the acronym PRCA or the term erythroblastopenia, pure red cell aplasia is characterised by severe anaemia with reticulocytopenia. It may occur in acute form induced by infectious agents, following drug toxicity or transplantation of allogeneic haematopoietic cells, associated with autoimmune haemolytic anaemia. The chronic form is rarely constitutional but can be acquired and is usually associated with blood or idiopathic diseases. IMMUNOLOGICAL INHIBITION OF ERYTHROPOIESIS: Among the mechanisms responsible for PRCA is immunological erythropoiesis inhibition. This may be of lymphocyte T cell origin or due to the presence of antibodies in the patient's serum. Although observations of PRCA with presence of neutralising antierythropoietin antibodies in patient's serum have multiplied over the past 5 years, they still remain extremely rare. From a therapeutic point of view, they require withdrawal of epoetin and often the administration of immunosuppressors and transfusion for symptomatic treatment. GROWTH FACTORS: The role of growth factors in restoring aplastic anaemia appears to be only partial, at random and temporary. PMID- 14534505 TI - [rHU-EPO in cancer patients. Target or wider use?]. AB - THE DEBATE: Although some believe that rHu-EPO should not be widely used in malignant affections, others think that because of the varied impact of these anaemia, its wider use should be recommended. FOR A TARGET USE: Various observations (influence of the degree of extension of the myelomas to the skeleton and response to specific treatment in the case of myeloma, whatever the haemoglobin concentration, degree of prevention of rHu-EPO chemo-induced anaemia) are in favour of its use in selected patients. FOR A WIDER USE: The benefits of treatment with rHu-EPO are not limited to the symptomatology of anaemia but extend to its potential complications in the most fragile patients. Other than the risks of infection, the heavy costs of transfusions must also be taken into account. PMID- 14534506 TI - [Epoetin beta, new strategies to optimise the management of anaemia in cancer patients]. PMID- 14534507 TI - Rosuvastatin--a new lipid-lowering drug. PMID- 14534508 TI - Lp-PLA2: a new marker of vascular risk. PMID- 14534509 TI - Cigarette smoking among adults--United States, 2001. AB - One of the national health objectives for the United States for 2010 is to reduce the prevalence of cigarette smoking among adults to > or =12% (objective 27.1a). To assess progress toward this objective, CDC analyzed self-reported data from the 2001 National Health Interview Survey (NHIS). The findings of this analysis indicate that, in 2001, approximately 22.8% of U.S. adults were current smokers compared with 25.0% in 1993. During 1965-2001, smoking prevalence declined faster among non-Hispanic blacks aged > or =18 years than among non-Hispanic whites the same age. Preliminary data for January-March 2002 indicate a continuing decline in current smoking prevalence among adults overall. However, the overall decline in smoking is not occurring at a rate that will meet the national health objective by 2010. Increased emphasis on a comprehensive approach to cessation that comprises educational, economic, clinical, and regulatory strategies is required to further reduce the prevalence of smoking in the United States. PMID- 14534510 TI - Yersinia enterocolitica gastroenteritis among infants exposed to chitterlings- Chicago, Illinois, 2002. AB - During December 2002-January 2003, the Chicago Department of Public Health (CDPH) investigated a cluster of Yersinia enterocolitica infections reported during a 10 week period among nine Chicago infants aged < or =1 year. This report summarizes the investigation of these cases and underscores the continuing risks for enteric infection among infants exposed to chitterlings (i.e., pork intestines), and the need for health-care providers to be aware of Y. enterocolitica as a cause of gastroenteritis, particularly in black children during traditional winter holiday celebrations. PMID- 14534511 TI - Racial/ethnic disparities in influenza and pneumococcal vaccination levels among persons aged > or =65 years--United States, 1989-2001. AB - Influenza and pneumococcal diseases are key causes of mortality among persons aged > or =65 years, accounting for approximately 36,000 and 3,400 deaths per year, respectively, during 1990-1999. Substantial racial/ethnic disparities in adult vaccination have been documented in national surveys. Although the national health objective for 2000 of 60% receipt of influenza vaccination during the preceding 12 months by persons aged > or =65 years (objective no. 20.11) was met in 1997, and the objective of 60% for pneumococcal vaccination was nearly met in 2000, vaccine coverage levels among non-Hispanic blacks and Hispanics were 31% and 30%, respectively, compared with 57% for non-Hispanic whites. To characterize these disparities, CDC analyzed data from the 2000 and 2001 National Health Interview Surveys (NHIS) and examined trends in NHIS results for 1989-2001. This report summarizes the results of these analyses, which indicate that marked differences in vaccination coverage by race/ethnicity are observed even among persons most likely to be vaccinated (e.g., persons with the highest education level and persons with frequent visits to health-care providers). Racial/ethnic disparities in influenza and pneumococcal vaccination coverage have persisted over time. Several approaches to reduce these disparities are needed, including increasing demand for vaccination among racial/ethnic minority populations and the use of standing orders and other systems changes that promote vaccination. PMID- 14534512 TI - Primary amebic meningoencephalitis--Georgia, 2002. AB - In early September 2002, the Georgia Division of Public Health and CDC were notified about a fatal case of primary amebic meningoencephalitis (PAM) caused by Naegleria fowleri in a boy aged 11 years who had recently swum in a local river. This report summarizes the case investigation. In response to this case, the district health department recommended that local community authorities advise persons to avoid swimming in this river during periods of high temperature and low water depth. PMID- 14534513 TI - West Nile virus activity--United States, October 2-8, 2003. AB - This report summarizes West Nile virus (WNV) surveillance data reported to CDC through ArboNET as of 3 a.m., Mountain Daylight Time, October 8, 2003. PMID- 14534514 TI - Diagnosis and treatment of autism spectrum disorders. PMID- 14534516 TI - The Gly389Arg beta1-adrenergic receptor polymorphism: a predictor of response to beta-blocker treatment? PMID- 14534517 TI - Respiratory and miotic effects of morphine in healthy volunteers when P glycoprotein is blocked by quinidine. AB - AIM: Our objective was to evaluate whether P-glycoprotein inhibition after quinidine pretreatment results in modified central nervous effects of morphine. METHODS: Twelve healthy volunteers received 7.5 mg morphine as an intravenous infusion over a 3-hour period. In a randomized, double-blind, 2-way crossover fashion, subjects received either 800 mg quinidine or placebo 1 hour before the start of morphine administration. The miotic and respiratory depressive effects of morphine were assessed by means of pupillometry and the respiratory response to carbon dioxide rebreathing, respectively. Quinidine effects were assessed by electrocardiogram recordings. Plasma concentrations of morphine and its glucuronide metabolites were measured throughout the observation period of 5 hours. RESULTS: Morphine significantly reduced both the respiratory response to carbon dioxide and the pupil diameter. Throughout the observation period, quinidine had significant effects on the corrected QT interval (QTc increase of >60 milliseconds), indicating clinically relevant quinidine action. However, quinidine pretreatment did not enhance the respiratory depressive effects of morphine, nor did it alter the miotic effects of morphine to a statistically significant or clinically relevant extent. Plasma concentrations of morphine and its glucuronides were not significantly changed by quinidine pretreatment. CONCLUSIONS: Whereas morphine clearly produced miosis and respiratory depression, pretreatment with quinidine as an inhibitor of P-glycoprotein did not result in an enhancement of central nervous opioid effects in healthy volunteers. PMID- 14534518 TI - Cisapride disposition in neonates and infants: in vivo reflection of cytochrome P450 3A4 ontogeny. AB - BACKGROUND: Cisapride, a prokinetic agent and substrate for cytochrome P450 (CYP) 3A4, has been used to treat neonates and infants with feeding intolerance and apnea or bradycardia associated with gastroesophageal reflux. At age 1 month, CYP3A4 activity has been reported to be only 30% to 40% of adult activity. This known developmental delay in the expression of CYP3A4 prompted us to conduct a classical open-label pharmacokinetic study of cisapride in neonates and young infants. METHODS: A total of 35 infants with a postconceptional age of 28 to 54 weeks at the time of the study received a single oral cisapride dose (0.2 mg/kg) at a postnatal age of 4 to 102 days, followed by repeated (n = 7) blood sampling over a 24-hour period. Cisapride and norcisapride were quantitated from plasma by HPLC-tandem mass spectrometry and pharmacokinetic data determined (n = 32) by noncompartmental methods. RESULTS: The pharmacokinetic parameters (mean +/- SD) were as follows: time to reach peak plasma concentration (t(max)), 4.4 +/- 2.8 hours (range, 0.9-12 hours); peak plasma concentration (C(max)), 29.3 +/- 16.6 ng/mL (range, 5.2-71.7 ng/mL); elimination half-life (t(1/2)), 10.7 +/- 3.7 hours (range, 1.9-18.1 hours); apparent total body clearance (Cl/F), 0.62 +/- 0.43 L. h(-1). kg(-1) (range, 0.2-1.9 L. h(-1). kg(-1)); and apparent volume of distribution (VD(ss)/F), 9.0 +/- 7.1 L/kg (range, 2.2-30.5 L/kg). The apparent renal clearance (CL(R)) of cisapride in infants (n = 28) was estimated to be 0.003 +/- 0.003 L. h(-1). kg(-1). Substratification of the population based on postconceptional age demonstrated the following findings for cisapride: (1) The mean (+/-SD) C(max) for cisapride was higher in the oldest postconceptional age category (44.5 +/- 19.6 ng/mL) than the middle and youngest categories (23.4 +/- 11.7 ng/mL and 30.0 +/- 17.5 ng/mL, respectively); (2) the t(max) for cisapride was shortest in the oldest postconceptional age category (2.2 +/- 1.1 hours) compared with the middle and youngest categories (4.4 +/- 3.3 hours and 5.0 +/- 2.6 hours, respectively); (3) the CL/F for cisapride in the youngest postconceptional age group was significantly lower (0.45 +/- 0.26 L. h(-1). kg( 1), P <.05) than in the middle and oldest categories (0.75 +/- 0.46 L. h(-1). kg( 1) and 0.85 +/- 0.69 L. h(-1). kg(-1), respectively); (4) a positive linear correlation was found between postconceptional age and the apparent terminal elimination rate constant (lambda(z)) for cisapride (P <.001, r(2) = 0.47) but not with CL/F. For norcisapride, the mean apparent C(max) was highest and the t(max) was shortest in the oldest postconceptional age group, although no association between postconceptional age and the norcisapride/cisapride area under the curve ratio was observed. All infants tolerated a single dose of cisapride well without significant alteration in QTc. CONCLUSIONS: (1) In neonates and infants, cisapride absorption and metabolism to its primary metabolite, norcisapride, were developmentally dependent; (2) approximately 99% of cisapride CL/F in neonates and young infants was nonrenal in nature; (3) CL/F of cisapride in neonates and infants noted in this study was reduced compared with data from older children and adults, likely as a result of developmental reductions in CYP3A4 activity; (4) as reflected by the correlation between postconceptional age and lambda(z), a rapid increase in total CYP3A4 activity occurs in the first 3 months of life. PMID- 14534519 TI - Inhibition of cytochrome P450 2B6 activity by hormone replacement therapy and oral contraceptive as measured by bupropion hydroxylation. AB - AIM: Our objective was to study the effect of hormone replacement therapy and a combination oral contraceptive on the cytochrome P450 (CYP) 2B6 activity with bupropion (INN, amfebutamone) hydroxylation used as a probe reaction. METHODS: This was a 3-way crossover study with 12 healthy female volunteers. The first phase was a control phase, in which all subjects received a single 150-mg dose of bupropion (sustained release) without pretreatment. In the second and third phases, in randomized balanced crossover order, subjects received a 10-day pretreatment with either hormone replacement therapy, containing 2 mg estradiol valerate and 250 microg levonorgestrel, or an oral contraceptive, containing 30 microg ethinyl estradiol (INN, ethinylestradiol) and 150 microg desogestrel, and the bupropion dose was given 1 hour after the last hormone dose. The bupropion, hydroxybupropion, and hydrobupropion plasma concentrations were determined for up to 72 hours. RESULTS: The 10-day hormone replacement therapy pretreatment reduced the hydroxybupropion/bupropion area under the plasma concentration-time curve (AUC) ratio by 49% (P <.001; 95% confidence interval [CI], -58% to -40%) as a result of a marked 47% decrease (P <.001; 95% CI, -54% to -41%) in the AUC of hydroxybupropion. Moreover, the AUC of hydrobupropion was significantly (64%; P =.003; 95% CI, 22% to 106%) increased. The AUC of hydroxybupropion was also reduced after the oral contraceptive treatment but to a lesser extent (-31%; P <.001; 95% CI, -37% to -26%). However, the hydroxybupropion/bupropion AUC ratio was not significantly affected. CONCLUSIONS: Hormone replacement therapy markedly inhibited the CYP2B6-catalyzed hydroxylation of bupropion, whereas a combination oral contraceptive had only a modest effect on CYP2B6 activity. Patients receiving hormone replacement therapy or oral contraceptives may need dose adjustment when treated with drugs metabolized by CYP2B6. PMID- 14534520 TI - Effect of rifampin on the pharmacokinetics and pharmacodynamics of gliclazide. AB - OBJECTIVE: Our objective was to investigate the effect of rifampin (INN, rifampicin) on the pharmacokinetics and pharmacodynamics of gliclazide, a sulfonylurea antidiabetic drug. METHOD: In a randomized 2-way crossover study with a 4-week washout period, 9 healthy Korean subjects were treated once daily for 6 days with 600 mg rifampin or with placebo. On day 7, a single dose of 80 mg gliclazide was administered orally. Plasma gliclazide, blood glucose, and insulin concentrations were measured. RESULTS: Rifampin decreased the mean area under the plasma concentration-time curve for gliclazide by 70% (P <.001) and the mean elimination half-life from 9.5 to 3.3 hours (P <.05). The apparent oral clearance of gliclazide increased about 4-fold after rifampin treatment (P <.001). A significant difference in the blood glucose response to gliclazide was observed between the placebo and rifampin phases. CONCLUSION: The effect of rifampin on the pharmacokinetics and pharmacodynamics of gliclazide suggests that rifampin affects the disposition of gliclazide in humans, possibly by the induction of cytochrome P450 2C9. Concomitant use of rifampin with gliclazide can considerably reduce the glucose-lowering effects of gliclazide. PMID- 14534521 TI - Pharmacokinetics of high doses of intramuscular and oral heroin in narcotic addicts. AB - BACKGROUND: In several countries medical prescription of diacetylmorphine is currently being evaluated as a treatment option for heavily dependent narcotic addicts. Because of damaged veins, many patients administer diacetylmorphine intramuscularly or orally. Therefore we characterized the pharmacokinetics of intramuscular and oral diacetylmorphine in the high dose range usually required in narcotic addicts. METHODS: Three intramuscular doses, 3 oral doses, and 1 intravenous dose of diacetylmorphine and oral and intravenous test doses of deuterium-labeled morphine (morphine-N-methyl-d3 [morphine-d3]) were administered to 8 heroin-addicted patients. Arterial plasma concentrations of diacetylmorphine, monoacetylmorphine, morphine, morphine-3-glucuronide, morphine 6-glucuronide, and morphine-d3 were measured by liquid chromatography-mass spectrometry. RESULTS: Intramuscularly administered diacetylmorphine ( 1, and the model also predicts temperature correlations in matching circles on the sky. PMID- 14534578 TI - Plant recognition of symbiotic bacteria requires two LysM receptor-like kinases. AB - Although most higher plants establish a symbiosis with arbuscular mycorrhizal fungi, symbiotic nitrogen fixation with rhizobia is a salient feature of legumes. Despite this host range difference, mycorrhizal and rhizobial invasion shares a common plant-specified genetic programme controlling the early host interaction. One feature distinguishing legumes is their ability to perceive rhizobial specific signal molecules. We describe here two LysM-type serine/threonine receptor kinase genes, NFR1 and NFR5, enabling the model legume Lotus japonicus to recognize its bacterial microsymbiont Mesorhizobium loti. The extracellular domains of the two transmembrane kinases resemble LysM domains of peptidoglycan- and chitin-binding proteins, suggesting that they may be involved directly in perception of the rhizobial lipochitin-oligosaccharide signal. We show that NFR1 and NFR5 are required for the earliest physiological and cellular responses to this lipochitin-oligosaccharide signal, and demonstrate their role in the mechanism establishing susceptibility of the legume root for bacterial infection. PMID- 14534580 TI - Fermi-liquid breakdown in the paramagnetic phase of a pure metal. AB - Fermi-liquid theory (the standard model of metals) has been challenged by the discovery of anomalous properties in an increasingly large number of metals. The anomalies often occur near a quantum critical point--a continuous phase transition in the limit of absolute zero, typically between magnetically ordered and paramagnetic phases. Although not understood in detail, unusual behaviour in the vicinity of such quantum critical points was anticipated nearly three decades ago by theories going beyond the standard model. Here we report electrical resistivity measurements of the 3d metal MnSi, indicating an unexpected breakdown of the Fermi-liquid model--not in a narrow crossover region close to a quantum critical point where it is normally expected to fail, but over a wide region of the phase diagram near a first-order magnetic transition. In this regime, corrections to the Fermi-liquid model are expected to be small. The range in pressure, temperature and applied magnetic field over which we observe an anomalous temperature dependence of the electrical resistivity in MnSi is not consistent with the crossover behaviour widely seen in quantum critical systems. This may suggest the emergence of a well defined but enigmatic quantum phase of matter. PMID- 14534581 TI - Temperature-induced valence transition and associated lattice collapse in samarium fulleride. AB - The different degrees of freedom of a given system are usually independent of each other but can in some materials be strongly coupled, giving rise to phase equilibria sensitively susceptible to external perturbations. Such systems often exhibit unusual physical properties that are difficult to treat theoretically, as exemplified by strongly correlated electron systems such as intermediate-valence rare-earth heavy fermions and Kondo insulators, colossal magnetoresistive manganites and high-transition temperature (high-T(c)) copper oxide superconductors. Metal fulleride salts-metal intercalation compounds of C60--and materials based on rare-earth metals also exhibit strong electronic correlations. Rare-earth fullerides thus constitute a particularly intriguing system--they contain highly correlated cation (rare-earth) and anion (C60) sublattices. Here we show, using high-resolution synchrotron X-ray diffraction and magnetic susceptibility measurements, that cooling the rare-earth fulleride Sm2.75C60 induces an isosymmetric phase transition near 32 K, accompanied by a dramatic isotropic volume increase and a samarium valence transition from (2 + epsilon) + to nearly 2 +. The negative thermal expansion--heating from 4.2 to 32 K leads to contraction rather than expansion--occurs at a rate about 40 times larger than in ternary metal oxides typically exhibiting such behaviour. We attribute the large negative thermal expansion, unprecedented in fullerene or other molecular systems, to a quasi-continuous valence transition from Sm(2+) towards the smaller Sm((2+epsilon)+), analogous to the valence or configuration transitions encountered in intermediate-valence Kondo insulators like SmS (ref. 3). PMID- 14534582 TI - Understanding and tuning the epitaxy of large aromatic adsorbates by molecular design. AB - If the rich functionality of organic molecules is to be exploited in devices such as light-emitting diodes or field-effect transistors, interface properties of organic materials with various (metallic and insulating) substrates must be tailored carefully. In many cases, this calls for well-ordered interfaces. Organic epitaxy-that is, the growth of molecular films with a commensurate structural relationship to their crystalline substrates--relies on successful recognition of preferred epitaxial sites. For some large pi-conjugated molecules ('molecular platelets') this works surprisingly well, even if the substrate exhibits no template structure into which the molecules can lock. Here we present an explanation for site recognition in non-templated organic epitaxy, and thus resolve a long-standing puzzle. We propose that this form of site recognition relies on the existence of a local molecular reaction centre in the extended pi electron system of the molecule. Its activity can be controlled by appropriate side groups and--in a certain regime--may also be probed by molecularly sensitized scanning tunnelling microscopy. Our results open the possibility of engineering epitaxial interfaces, as well as other interfacial nanostructures for which specific site recognition is essential. PMID- 14534583 TI - Growth of early continental crust by partial melting of eclogite. AB - The tectonic setting in which the first continental crust formed, and the extent to which modern processes of arc magmatism at convergent plate margins were operative on the early Earth, are matters of debate. Geochemical studies have shown that felsic rocks in both Archaean high-grade metamorphic ('grey gneiss') and low-grade granite-greenstone terranes are comprised dominantly of sodium-rich granitoids of the tonalite-trondhjemite-granodiorite (TTG) suite of rocks. Here we present direct experimental evidence showing that partial melting of hydrous basalt in the eclogite facies produces granitoid liquids with major- and trace element compositions equivalent to Archaean TTG, including the low Nb/Ta and high Zr/Sm ratios of 'average' Archaean TTG, but from a source with initially subchondritic Nb/Ta. In modern environments, basalts with low Nb/Ta form by partial melting of subduction-modified depleted mantle, notably in intraoceanic arc settings in the forearc and back-arc regimes. These observations suggest that TTG magmatism may have taken place beneath granite-greenstone complexes developing along Archaean intraoceanic island arcs by imbricate thrust-stacking and tectonic accretion of a diversity of subduction-related terranes. Partial melting accompanying dehydration of these generally basaltic source materials at the base of thickened, 'arc-like' crust would produce compositionally appropriate TTG granitoids in equilibrium with eclogite residues. PMID- 14534584 TI - Large Cretaceous sphenodontian from Patagonia provides insight into lepidosaur evolution in Gondwana. AB - Sphenodontian reptiles successfully radiated during Triassic and Jurassic times, but were driven almost to extinction during the Cretaceous period. The sparse Early Cretaceous record of sphenodontians has been interpreted as reflecting the decline of the group in favour of lizards, their suspected ecological successors. However, recent discoveries in Late Cretaceous beds in Patagonia partially modify this interpretation. Numerous skeletons of a new sphenodontian, Priosphenodon avelasi gen. et sp. nov., were collected from a single locality in the Cenomanian Turonian Candeleros Formation, where it is more abundant than any other tetrapod group recorded in the quarry (for example, Crocodyliformes, Serpentes, Dinosauria and Mammalia). Adult specimens of Priosphenodon reached one metre in length, larger than any previously known terrestrial sphenodontian. Here we propose, using available evidence, that sphenodontians were not a minor component of the Cretaceous terrestrial ecosystems of South America, and that their ecological replacement by squamates was delayed until the early Tertiary. The new discovery helps to bridge the considerable gap in the fossil record (around 120 million years) that separates the Early Cretaceous sphenodontians from their living relatives (Sphenodon). PMID- 14534585 TI - Ultraviolet vision in a bat. AB - Most mammals, with the exception of primates, have dichromatic vision and correspondingly limited colour perception. Ultraviolet vision was discovered in mammals only a decade ago, and in the few rodents and marsupials where it has been found, ultraviolet light is detected by an independent photoreceptor. Bats orient primarily by echolocation, but they also use vision. Here we show that a phyllostomid flower bat, Glossophaga soricina, is colour-blind but sensitive to ultraviolet light down to a wavelength of 310 nm. Behavioural experiments revealed a spectral-sensitivity function with maxima at 510 nm (green) and above 365 nm (ultraviolet). A test for colour vision was negative. Chromatic adaptation had the same threshold-elevating effects on ultraviolet and visible test lights, indicating that the same photoreceptor is responsible for both response peaks (ultraviolet and green). Thus, excitation of the beta-band of the visual pigment is the most likely cause of ultraviolet sensitivity. This is a mechanism for ultraviolet vision that has not previously been demonstrated in intact mammalian visual systems. PMID- 14534586 TI - Consolidation during sleep of perceptual learning of spoken language. AB - Memory consolidation resulting from sleep has been seen broadly: in verbal list learning, spatial learning, and skill acquisition in visual and motor tasks. These tasks do not generalize across spatial locations or motor sequences, or to different stimuli in the same location. Although episodic rote learning constitutes a large part of any organism's learning, generalization is a hallmark of adaptive behaviour. In speech, the same phoneme often has different acoustic patterns depending on context. Training on a small set of words improves performance on novel words using the same phonemes but with different acoustic patterns, demonstrating perceptual generalization. Here we show a role of sleep in the consolidation of a naturalistic spoken-language learning task that produces generalization of phonological categories across different acoustic patterns. Recognition performance immediately after training showed a significant improvement that subsequently degraded over the span of a day's retention interval, but completely recovered following sleep. Thus, sleep facilitates the recovery and subsequent retention of material learned opportunistically at any time throughout the day. Performance recovery indicates that representations and mappings associated with generalization are refined and stabilized during sleep. PMID- 14534587 TI - Dissociable stages of human memory consolidation and reconsolidation. AB - Historically, the term 'memory consolidation' refers to a process whereby a memory becomes increasingly resistant to interference from competing or disrupting factors with the continued passage of time. Recent findings regarding the learning of skilled sensory and motor tasks ('procedural learning') have refined this definition, suggesting that consolidation can be more strictly determined by time spent in specific brain states such as wake, sleep or certain stages of sleep. There is also renewed interest in the possibility that recalling or 'reactivating' a previously consolidated memory renders it once again fragile and susceptible to interference, therefore requiring periods of reconsolidation. Using a motor skill finger-tapping task, here we provide evidence for at least three different stages of human motor memory processing after initial acquisition. We describe the unique contributions of wake and sleep in the development of different forms of consolidation, and show that waking reactivation can turn a previously consolidated memory back into a labile state requiring subsequent reconsolidation. PMID- 14534588 TI - Environmentally mediated synergy between perception and behaviour in mobile robots. AB - The notion that behaviour influences perception seems self-evident, but the mechanism of their interaction is not known. Perception and behaviour are usually considered to be separate processes. In this view, perceptual learning constructs compact representations of sensory events, reflecting their statistical properties, independently of behavioural relevance. Behavioural learning, however, forms associations between perception and action, organized by reinforcement, without regard for the construction of perception. It is generally assumed that the interaction between these two processes is internal to the agent, and can be explained solely in terms of the neuronal substrate. Here we show, instead, that perception and behaviour can interact synergistically via the environment. Using simulated and real mobile robots, we demonstrate that perceptual learning directly supports behavioural learning and so promotes a progressive structuring of behaviour. This structuring leads to a systematic bias in input sampling, which directly affects the organization of the perceptual system. This external, environmentally mediated feedback matches the perceptual system to the emerging behavioural structure, so that the behaviour is stabilized. PMID- 14534589 TI - Regulation of neuroblast competence in Drosophila. AB - Individual neural progenitors generate different cell types in a reproducible order in the retina, cerebral cortex and probably in the spinal cord. It is unknown how neural progenitors change over time to generate different cell types. It has been proposed that progenitors undergo progressive restriction or transit through distinct competence states; however, the underlying molecular mechanisms remain unclear. Here we investigate neural progenitor competence and temporal identity using an in vivo genetic system--Drosophila neuroblasts--where the Hunchback transcription factor is necessary and sufficient to specify early-born cell types. We show that neuroblasts gradually lose competence to generate early born fates in response to Hunchback, similar to progressive restriction models, and that competence to acquire early-born fates is present in mitotic precursors but is lost in post-mitotic neurons. These results match those observed in vertebrate systems, and establish Drosophila neuroblasts as a model system for the molecular genetic analysis of neural progenitor competence and plasticity. PMID- 14534590 TI - The Wnt/beta-catenin pathway regulates cardiac valve formation. AB - Truncation of the tumour suppressor adenomatous polyposis coli (Apc) constitutively activates the Wnt/beta-catenin signalling pathway. Apc has a role in development: for example, embryos of mice with truncated Apc do not complete gastrulation. To understand this role more fully, we examined the effect of truncated Apc on zebrafish development. Here we show that, in contrast to mice, zebrafish do complete gastrulation. However, mutant hearts fail to loop and form excessive endocardial cushions. Conversely, overexpression of Apc or Dickkopf 1 (Dkk1), a secreted Wnt inhibitor, blocks cushion formation. In wild-type hearts, nuclear beta-catenin, the hallmark of activated canonical Wnt signalling, accumulates only in valve-forming cells, where it can activate a Tcf reporter. In mutant hearts, all cells display nuclear beta-catenin and Tcf reporter activity, while valve markers are markedly upregulated. Concomitantly, proliferation and epithelial-mesenchymal transition, normally restricted to endocardial cushions, occur throughout the endocardium. Our findings identify a novel role for Wnt/beta catenin signalling in determining endocardial cell fate. PMID- 14534594 TI - Young scientist: Italian biotechnologists organize. PMID- 14534595 TI - Nuts and bolts. PMID- 14534591 TI - A receptor kinase gene of the LysM type is involved in legume perception of rhizobial signals. AB - Plants belonging to the legume family develop nitrogen-fixing root nodules in symbiosis with bacteria commonly known as rhizobia. The legume host encodes all of the functions necessary to build the specialized symbiotic organ, the nodule, but the process is elicited by the bacteria. Molecular communication initiates the interaction, and signals, usually flavones, secreted by the legume root induce the bacteria to produce a lipochitin-oligosaccharide signal molecule (Nod factor), which in turn triggers the plant organogenic process. An important determinant of bacterial host specificity is the structure of the Nod-factor, suggesting that a plant receptor is involved in signal perception and signal transduction initiating the plant developmental response. Here we describe the cloning of a putative Nod-factor receptor kinase gene (NFR5) from Lotus japonicus. NFR5 is essential for Nod-factor perception and encodes an unusual transmembrane serine/threonine receptor-like kinase required for the earliest detectable plant responses to bacteria and Nod-factor. The extracellular domain of the putative receptor has three modules with similarity to LysM domains known from peptidoglycan-binding proteins and chitinases. Together with an atypical kinase domain structure this characterizes an unusual receptor-like kinase. PMID- 14534597 TI - Theory of functional systems and human general pathology. AB - We analyze the role of the theory of functional systems for human general pathology and the necessity of integration of this theory with the concepts of pathological and ambivalent systems. Multiple (qualitatively heterogeneous) nature of system-forming factors and principle possibility of the formation of physiological, pathological, and ambivalent systems by the same factors are discussed. These theses broaden the application of the theory of functional systems as the fundamental basis for studies of informational mechanisms of vital activity under normal and pathological conditions. PMID- 14534598 TI - Role of nitric oxide and lipid peroxidation in pathophysiological mechanisms of audiogenic seizures in GEP Rats and DBA/2 mice. AB - We evaluated the role of nitric oxide and lipid peroxidation in the pathophysiological mechanisms of seizures in genetically epilepsy prone (GEP) rats and DBA/2 mice with genetically determined audiogenic epilepsy. In rats and mice acoustic stimulation led to locomotor activation followed by clonic-tonic seizures. The contents of nitric oxide and lipid peroxidation products at the peak of seizures markedly surpassed the control level. PMID- 14534599 TI - Effect of transitory ischemia on liver lysosomal apparatus in rats with different resistance to hypoxia. AB - We studied the state of lysosomal apparatus and pro- and antioxidant activity in the liver of rats with different resistance to hypoxia during postischemic recovery. Under normal conditions the lysosomal apparatus did not differ in highly and low resistant animals. During ischemia and reperfusion the damage to hepatic lysosomal membranes in rats highly resistant to hypoxia was less pronounced than in low resistant animals. These differences also concerned labilization of lysosomes during exposure to damaging factors (hypotonia and Triton X-100). The rats highly resistant to hypoxia differed from low resistant animals by higher stability of lysosomal membranes, lower prooxidant activity (malonic dialdehyde content), and higher tissue concentration of alpha-tocopherol during reperfusion. PMID- 14534600 TI - Effect of physical training on blood level of endogenous modulators of beta adreno- and m-cholinoreactivity in patients with a history of myocardial infarction. AB - The relative content of myocyte-active factors and endogenous muscarinic receptor blocker in the blood increased, while the concentration of endogenous beta adrenoceptor sensitizer decreased in coronary patients with a history of acute myocardial infarction. Physical training produced a therapeutic effect, normalized the content of these factors and, probably, improved beta-adrenergic and muscarinic cholinergic regulation of the heart and vessels. PMID- 14534601 TI - Alteration of the liver in rats with experimental dysbiosis. AB - We evaluated the relationship between pathological changes in the liver and the state of intestinal microflora in rats with experimental dysbiosis. Changes in the intestinal microflora were accompanied by alteration of the morphological structure in the liver. Enhanced proliferation of Ito cells served as an indirect evidence of damage to the liver. Ito cells did not undergo transformation into myofibroblasts that excluded the possibility of fibrosis. PMID- 14534602 TI - Effect of chronic stress during early postnatal ontogeny on structural characteristics of the myocardium and glomerular apparatus of the kidneys in NISAG rats. AB - We studied structural characteristics of the myocardium and glomerular apparatus of the kidneys in 3-week- and 6-month-old male NISAG rats (hereditary stress induced arterial hypertension) subjected to handling on days 1-21 of postnatal ontogeny. The animals were daily isolated from mothers for 10 min. Handling did not modulate the development of arterial hypertension and typical morphological signs in the myocardium and kidneys. PMID- 14534603 TI - Detection and evaluation of NO stores in awake rats. AB - We demonstrated and substantiated the possibility of detection and evaluation of NO stores in freely moving awake rats. NO stores were created by administering NO donor or by heat shock and were then detected by hypotensive reaction to diethyldithiocarbamate (blood pressure monitoring) under conditions of NO synthase inhibition. Electron paramagnetic resonance revealed NO release from its stores by incorporation into paramagnetic mononitrosyl-iron complexes with diethyldithiocarbamate. Five hours after administration of NO donor or heat shock diethyldithiocarbamate induced a blood pressure drop and the appearance of electron paramagnetic resonance signals from the mononitrosyl-iron diethyldithiocarbamate complex in rat heart, liver, kidneys, and brain. The hypotensive reaction to diethyldithiocarbamate and electron paramagnetic resonance signals were absent in control rats. PMID- 14534604 TI - Effect of transmural pressure on constrictor reactions of caudal artery in hypotensive and hypertensive rats. AB - Clipping of the abdominal aorta distally to the renal arteries produces a persistent decrease in blood pressure in hindquarter vessels by 35-40%. On week 6 7 postoperation, the reactions of the caudal artery perfused in vitro under constant pressure to norepinephrine were studied. At transmural pressure of 150 mm Hg, the vascular responses in hypotensive rats were reduced compared to those in normotensive control. By contrast, the responses of hypertensive vessels were more pronounced at 75 mm Hg even after deendothelization. PMID- 14534605 TI - Parameters of adenylate pool as predictors of energy metabolism disturbances in hepatocytes during hypoxia. AB - We studied the dependence of various parameters of adenylate pool in hepatocytes on oxygen concentrations. Isolated cells responded to a decrease in oxygen content in their microenvironment by changes in components of the adenine nucleotide system, which attested to phasic nature of this process. Three ranges of oxygen concentrations differing by the type of changes in the parameters of adenylate pool were distinguished: steady-state range of these parameters; primary changes in the adenylate pool aimed at minimization of energy losses (compensatory stage characteristic of the initial stages of hypoxia); and linear drop of ATP content paralleled by decompensation of the regulatory mechanisms of ATP formation and adenine nucleotide degradation. Hence, parameters of the adenylate pool can serve as predictors of different stages of hypoxia. Differences in the parameters of adenylate pool depending on the level of O(2) in hepatocytes of rats highly and low-resistant to hypoxia indicate that energy metabolism is a mechanism involved in the formation of individual cell resistance to oxygen deficiency. These data suggest that suspension of isolated hepatocytes as an adequate cellular model for experimental studies of the effects of hypoxia on energy metabolism and functional activity of the cell. PMID- 14534606 TI - Dose-dependent effect of alpha-tocopherol on activity of xenobiotic metabolizing enzymes in rat liver. AB - Oral treatment with alpha-tocopherol for 4 days dose-dependently increased the content of cytochrome P450 (CYP), catalytic activities of CYP1A1, CYP1A2, CYP2B1, CYP2C, and activity of NADPH-cytochrome-P450 reductase in the liver of male rats, but did not change activity of glutathione S-transferase. These results suggest that alpha-tocopherol induced the enzymes of phase I of xenobiotic metabolism, including CYP1 and CYP2 families involved in the metabolism of drugs and procarcinogenes. PMID- 14534607 TI - Oxidation of plasma low-density lipoproteins from coronary patients with various forms of hypercholesterolemia. AB - The duration of lag-phase of copper-induced free-radical oxidation of atherogenic LDL isolated from the plasma of coronary patients without hypercholesterolemia virtually does not differ from that of normal human LDL. On the other hand, lag phase of plasma LDL oxidation was minimal in coronary patients with primary hypercholesterolemia without familial history and especially in patients with familial hypercholesterolemia. This can be attributed to sharply decreased content of natural lipid antioxidants in LDL of patients with familial hypercholesterolemia. However, therapy with natural antioxidant vitamin E did not modulate oxidizability of these LDL. By contrast, therapy with beta-hydroxy-beta methylglutaryl-coenzyme A reductase inhibitor suppressing biosynthesis of ubiphenol Q induced sharp accumulation of lipoperoxides in LDL in vivo. These data suggest that reduced form of ubiquinone Q is the main antioxidant protecting LDL from free-radical oxidation. PMID- 14534608 TI - Cystein proteinase inhibitor stefin A as an indicator of efficiency of tumor treatment in mice. AB - The concentration of stefin A (cystatin A in mice) was measured in animals with experimental tumors (LS lymphosarcoma, HA-1-hepatoma, and Lewis lung carcinoma) during effective antitumor therapy. In mice with these tumors serum concentrations of stefin A increased, while the concentration of cystatin C (extracellular cystein proteinase inhibitor) decreased. The concentration of stefin A in tumor tissue in Lewis lung carcinoma was higher than in LS lymphosarcoma and HA-1-hepatoma ascitic cells, which can be explained by the degree of their malignancy. The content of stefin A in tumor tissue was similar to that in the liver and spleen of tumor-bearing animals, while its concentration in the liver and spleen of tumor-bearing animals was lower than in intact mice. The level of stefin A is an important marker of malignancy and an indicator of the efficiency of antitumor therapy. PMID- 14534609 TI - Content of autoantibodies to bradykinin and beta-amyloid(1-42) as a criterion for biochemical differences between Alzheimer's dementias. AB - We measured serum content of autoantibodies to beta-amyloid protein Abeta(1-42), its neurotoxic fragment Abeta(25-35), vasopressin, bradykinin, thrombin, antithrombin III, alpha(2)-macroglobulin, and angiotensin II in patients with various forms of Alzheimer's dementias, including presenile and senile dementias of the Alzheimer type. The ratio of antibradykinin and anti-Abeta(1-42) autoantibody contents differed by 39% in these patients. Our results can be used for the development of a new biochemical method for differential diagnostics of dementias of the Alzheimer type. PMID- 14534610 TI - Mechanisms underlying combined action of clonidine and local anesthetics. AB - We studied the mechanisms of combined action of clonidine and local anesthetics amethocaine and imidazo-benzimidazole derivative RU-1117. In contrast to amethocaine, RU-1117 in therapeutic concentrations binds to imidazoline receptors and, to a lesser extent, to alpha(2A)-adrenoceptors on human platelets. Clonidine and RU-1117 produce opposite effects on platelet aggregation induced by ADP in low concentrations. Our results suggest that the pharmacodynamic interaction of clonidine and RU-1117 is associated with their ability to activate the imidazolinergic system. PMID- 14534611 TI - Drug correction of behavioral reactions and metabolic disorders in rats with craniocerebral trauma. AB - Intraperitoneal injection of bemithyl in a dose of 25 mg/kg for 3 days after craniocerebral injury reduced psychopathological symptoms in rats with different resistance to acute hypoxia, restored the structure of individual behavior, and prevented metabolic disorders in the brain. PMID- 14534612 TI - Effect of pyroglutamylasparagine amide on plastic characteristics of synaptic transmission in the hippocampus. AB - Preincubation of rat hippocampal slices with 0.05-0.5 microM pyroglutamylasparagine amide improved characteristics of long-term potentiation of focal responses in the synaptic system of Schaffer collaterals-CA1 field pyramids facilitating LTP development and increasing its amplitude and duration. Presumably, the positive modulation of plastic characteristics of synaptic transmission in the hippocampusis is responsible for facilitation of learning and memory induced by pyroglutamylasparagine. PMID- 14534613 TI - Therapeutic efficiency of dimephosphone and xydiphone in experimental pulse therapy with prednisolone. AB - Experiments on rats showed that pulse therapy with prednisolone (100 mg/kg intraperitoneally for 3 days) stimulated urinary excretion of hydroxyproline, increased the content of inorganic phosphorus, promoted the increase in the content of dienic conjugates and catalase activity, and decreased serum levels of MDA and ceruloplasmin. Ten-day treatment with dimephosphone (208 mg/kg) or xydiphone (45 mg/kg) after pulse therapy with prednisolone normalized urinary excretion of hydroxyproline and reduced the levels of dienic conjugates. Dimephosphone did not change, while xydiphone normalized the level of MDA decreased by prednisolone. PMID- 14534614 TI - Effect of lycopine on the resistance of rat liver microsomes to in vitro induced LPO. AB - Lycopine in concentrations of 0.5-50 microM suppressed LPO in microsomes induced by NADPH-Fe(2+) and by ascorbic acid-Fe(2+). Lycopine in a concentration of 20 microM completely prevented the decrease in the rate of benz[a]pyrene hydroxylation and activation of p-nitrophenyl-UDP-glucuronosyl transferase caused by LPO induction in microsomes. PMID- 14534615 TI - Immunostimulating activity of synthetic bursopeptides. AB - Biologically active peptides bursopeptide-1 (Tyr-Glu-Glu) and bursopeptide-2 (Trp Thr-Ala-Glu-Glu-Lys-Gln-Leu) from the bursa of Fabricius were isolated and synthesized. Bursopeptide-1 stimulated expression of B cell differentiation antigens in lymphocyte culture from patients with burns. Bursopeptide-2 produced a variety of effects and stimulated expression of differentiation antigens on T and B cells and natural killer cells. Bursopeptides can modulate the mechanisms of interleukin-2-mediated lymphocyte activation. Bursopeptides stimulated the immune response in mice with cyclophosphamide-induced immunodeficiency, which manifested in increased hemagglutinin and hemolysin titers and increased count of antibody-producing cells in the spleen. PMID- 14534616 TI - Relationship between glutathione S-transferase P1 polymorphism and bronchial asthma and atopic dermatitis. AB - We determined the prevalence of GSTP1-Ile105 and GSTP1-Val105 alleles in patients with bronchial asthma and atopic dermatitis and healthy children of 2 groups (randomized and nonatopic control). The GSTP1-Ile105/Val105 genotype determines the resistance to atopic dermatitis (odds ratio=0.51; 95% confidence interval: 0.28-0.92; p=0.023). However, both homozygotes are at high risk of developing atopic dermatitis (near-significant differences). PMID- 14534618 TI - Amyloidosis in macaques in adler primatological center. AB - Results of pathomorphological studies of 133 amyloidosis cases in macaques of 3 species (M. mulatta, M. nemestrina, and M. fascicularis) in the Adler Primatological Center are presented. A relationship between the development of amyloidosis and animal age is detected and the incidence of involvement of various organs is determined. Generalized and isolated forms of amyloidosis are described, the most incident of which (36.8%) was hepatic amyloidosis. Simian and human amyloidosis are compared. Amyloidosis in macaques can be used as the model process for studies of amyloidosis in humans. PMID- 14534617 TI - Effects of etoposide and fludarabine in subtoxic doses on karyotype of human malignant lymphoid Namalwa cells. AB - We studied changes in the karyotype of transplanted Namalwa cells induced by DNA damaging antitumor preparations etoposide and fludarabine in subtoxic doses. The relative number of cells containing increased number of chromosomes and the incidence of chromatid aberrations with primary damage to chromosomes 2, 5, 11, 16, and 17 increased. Cytogenetic changes developed even after short-term incubation of cells with antitumor preparations and were observed during further culturing in a medium not containing etoposide or fludarabine. PMID- 14534619 TI - Labeled citrate metabolism in bone fractures and impaired innervation. AB - Changes in (14)C incorporation into regenerate after bone fracture and impairment of mandibular innervation, and injection of [3-(14)C]cytrate corresponded to the stages of reparative osteogenesis: after 1 week (14)C incorporation in the cellular-fibrous callus surpassed its release, after 2 weeks the rates of (14)C incorporation and release in the chondroid callus become similar, and after 4 weeks the release of the label predominated in the primary bone callus. Denervation reduced (14)C incorporation into regenerate, which impaired bone remodeling. Citrate in the bones is characterized by high metabolic activity. PMID- 14534620 TI - Xenotransplantation of embryonic precursors of human myogenesis for the correction of dystrophinopathy in mice with hereditary muscular dystrophy. AB - Human embryonic myogenic precursors were transplanted into muscles of mdx mice with hereditary dystrophin-deficient muscular dystrophy. Transplantation induced the synthesis of human dystrophin. The number of dystrophin-positive fibers progressively decreased, however, some of them were preserved even 5 months after transplantation. Our results indicate that xenogeneic transplantation of embryonic myogenic precursors compensates the genetic defect in dystrophin deficient mice. PMID- 14534621 TI - Pathomorphological criteria of arrhythmogenic heart. AB - Pathomorphological criteria of arrhythmogenic heart include structural compartmentalization with primary changes in the right ventricle and interventricular septum, fibro- and lipomatosis of the myocardium, and disseminated coronary obstruction. Ischemic focuses in the conducting system are the site of formation of arrhythmogenic substance promoting the development and progression of cardiac arrhythmias. Cardioneuropathy and pathological motility of the interventricular septum lead to systolic dysfunction and contribute to asynchronous excitation and contraction of ventricles in arrhythmogenic heart. PMID- 14534622 TI - Morphocytochemical assay of neuronal populations in cerebellar nuclei in birds and mammals of some ecological and morphological groups. AB - We performed a quantitative morphocytochemical study of neuronal populations in the lateral cerebellar nucleus in birds and mammals of some ecological and morphological groups. Morphological parameters of neuronal density, linear values, and structural nuclear-cytoplasmic index of cells were compared. Specific features of neuronal proteins in cerebellar nuclei were revealed. We revealed objective criteria for adaptive capacities at the level of cell populations in the lateral cerebellar nuclei of animals adapted to various environmental conditions. Our results extend the notion about morphological characteristics of synanthropic birds and rodents that carry infections and invasions hazardous to humans. PMID- 14534623 TI - Ultrasound as a tool in experimental research of macro- and microcirculation. AB - Peculiarities of application of two modes of ultrasonic measurements of blood flow in experimental research are considered, which are based on Doppler effect and on differential transit-time of upstream and downstream sound propagation. The efficiency of high-frequency ultrasound flowmeter equipped with 26.8 MHz transducers was demonstrated in measurements of blood flow in rat midbrain and coronary arteries. This approach can be used for evaluation of the dynamics of cardiac output with an intravascular catheter 0.6 mm in diameter working at 33 MHz. The probe and electronic scheme of the devise for measuring blood flow in microvessels are described. Blood flow rate measured in mesentery and m. cremaster arterioles under normal conditions was 2-12 mm/sec. One-element probe working at 38.5 MHz provided stable recording of blood microflows in 30-40-micro vessels. PMID- 14534625 TI - ICT in health care: sociotechnical approaches. AB - The importance of the social sciences for medical informatics is increasingly recognized. As ICT requires inter-action with people and thereby inevitably affects them, understanding ICT requires a focus on the interrelation between technology and its social environment. Sociotechnical approaches increase our understanding of how ICT applications are developed, introduced and become a part of social practices. Socio-technical approaches share several starting points: 1) they see health care work as a social, 'real life' phenomenon, which may seem 'messy' at first, but which is guided by a practical rationality that can only be overlooked at a high price (i.e. failed systems). 2) They see technological innovation as a social process, in which organizations are deeply affected. 3) Through in-depth, formative evaluation, they can help improve system design and implementation. PMID- 14534624 TI - Use of isopropyl alcohol in histological assays: dehydration of tissue, enbessing into paraffin, and processing of paraffin sections. AB - Isopropyl alcohol (isopropanol) can be used as a substitute for ethyl alcohol in tissue dehydration during embedding into paraffin and dehydration of stained sections. The use of isopropyl alcohol during paraffinization allows us to exclude treatment with intermediate solvents of paraffin (chloroform, xylene, and benzene), which reduces the degree of tissue compaction and simplifies and accelerates histological assay. PMID- 14534626 TI - Medical image quality as a socio-technical phenomenon. AB - OBJECTIVES: The study aims to interpret image quality in laparoscopic surgery not only as a technical parameter but also as the result of the situation of use. METHODS: Observational studies of laparoscopy in use, semi-structured and informal interviews with laparoscopists. RESULTS: When medical images are digitized to exploit novel technical possibilities, image quality becomes a paramount issue. Image quality is often discussed exclusively in technical terms, but the socio-technical study of image quality in surgical telemedicine presented in this paper showed that it is definitely more than a purely technical parameter. CONCLUSIONS: While the resulting quality of the image was significantly shaped by the persons involved, the concept of "quality" itself was also relative and changing with the situation of use. A given technology does not determine image quality. Rather than focusing only on the technical quality, the attention of designers and decision makers should also be directed to the socio technical network surrounding the image and its use. PMID- 14534627 TI - Notions of reliability: considering the importance of difference in guiding patients to health care Web sites. AB - OBJECTIVES: This article analyzes the efforts of three organizations to provide a standard that guides Internet users to reliable health care sites. METHODS: Comparison of health Internet sites, interviews and document studies. RESULTS: In comparing these approaches, three different constructions of reliability are identified. CONCLUSIONS: The resulting possibilities and restrictions of these constructions for users that are searching for health information on the Internet are revealed. PMID- 14534628 TI - Perceptions of physician order entry: results of a cross-site qualitative study. AB - OBJECTIVE: To identify perspectives of success factors for implementing computerized physician order entry (POE) in the inpatient setting. DESIGN: Qualitative study by a multidisciplinary team using data from observation, focus groups, and both formal and informal interviews. Data were analyzed using a grounded approach to develop a taxonomy of patterns and themes from the transcripts and field notes. RESULTS: A taxonomy of ten high level themes was developed, including 1) separating POE from other processes, 2) terms, concepts, and connotations, 3) context, 4) tradeoffs, 5) conflicts and contradictions, 6) collaboration and trust, 7) leaders and bridgers, 8) the organization of information, 9) the ongoing nature of implementation, and 10) temporal concerns. CONCLUSION: The identified success factors indicate that POE implementation is an iterative and difficult process, but informants perceive it is worth the effort. PMID- 14534629 TI - Getting the big picture: the macro-politics of information system development (and failure) in a Canadian hospital. AB - OBJECTIVES: While recognized that global actors influence health information system design, studies of health informatics have largely focused on micro politics of technology design and implementation. Here a problematic patient care information system (PCIS) is discussed in relation to federal and provincial policies and corporate strategies to demonstrate that our understanding of health informatics can be enhanced by linking micro studies of health informatics to larger macro contexts. METHODS: Interviews and document study. RESULTS: Although the extent to which federal initiatives influenced (or failed to influence) provincial and hospital initiatives remains debateable, events initiated at one level (the hospital's decision to implement software, initiated at the organizational level) are influenced (perhaps indirectly) by developments in other contexts (federal/macro changes gave an initiative more weight; provincial initiatives such as the Labour Accord altered the industrial relations environment in which system development occurred). CONCLUSIONS: Micro-studies of work practice, invaluable in addressing interactions between technologies, users and work practices, often fail to account for the historic reach of global actors, although it is often these historic circumstances that contribute to present-day interactions between user, information system and organization, and that find expression - often indirectly - in daily work practices. PMID- 14534630 TI - Cognitive tools in medical teamwork: the spatial arrangement of patient records. AB - OBJECTIVES: As a preliminary for the design of Computer-Based Patient Records, the aim of this paper is to build an understanding of the roles physical artifacts like paper-based patient records play in supporting cognition and collaboration in the health-care settings. METHOD: A small ethnographically informed study was conducted in the emergency room at a 250-bed hospital in Sweden from the perspective of Distributed Cognition. RESULTS: To track work-in progress, clinicians placed patient records on a desk to form a shared public display that represented the current problem state for the health-care team. The results of the study suggest that the patient records and other physical artifacts are used by clinicians in different ways to form cognitive tools that offload memory tasks and support joint attention and collaboration. CONCLUSION: To design Computer-Based Patient Records that more appropriately support cognition and teamwork, it is important to investigate how clinicians make use of the paper-based patient records. Practitioners take advantage of existing tools frequently to deal with cognitively demanding tasks and collaboration issues. PMID- 14534631 TI - The search for synergy: interrelating medical work and patient care information systems. AB - OBJECTIVES: This article aims to search for the way patient care information systems can be most fruitfully put to work in health care. METHODS: Given the calls for improved health care quality, there is a need for IT's coordination capacities in health care. Yet IT's track record in this area is less than many expected. Moreover, many argue that the nature of health care work sets natural limits to the possibilities of IT to revolutionize this work. Starting with an analysis of the paper record, this article explores the way IT and professional work can be interrelated synergistically. RESULTS: Two principles are discussed: 1) The key to a fruitful operation of IT in health care work lies in the unraveling of the care process, and the redistribution of tasks between professionals and the IT application. 2) Professionals should be given the skills and resources to adapt the IT application's demands to the needs of their work practices. CONCLUSION: IT can bring true process support to health care when taking the two principles discussed here into account. PMID- 14534632 TI - Trusting the record. AB - OBJECTIVES: This paper aims to contribute to a longstanding interest in documents and paperwork in healthcare work through an examination of everyday work with patient records in a clinic. METHODS: An ethnographic study of record keeping practices in a deliberate self harm clinic was conducted to consider the role that document work plays in the development of trust in the routine social interactions of a working division of labor. RESULTS AND CONCLUSIONS: Issues of trust are seen to play central roles within the complexities of organizational working and some consequent implications for the deployment and use of electronic medical record systems are considered. PMID- 14534633 TI - Images of health technology in national and local strategies. AB - OBJECTIVES: This paper examines the potential of various models relating technology to society and institutional structures to inform health policy. Among the models discussed are various versions of technological determinism, social constructivism, actor network theory and critical theory. METHODS: The paper considers recent developments in policy and strategy that aim to shape the way the UK's National Health Service (NHS) integrates Information and Communication Technologies (ICTs) into health care and considers what these alternative models highlight or emphasise, and how they might influence the activities of setting local implementation strategies. RESULTS AND CONCLUSIONS: Contemporary ICTs are often presented as having a particular relevance and power in reforming or transforming the delivery of health care. Understanding how such technologies might be conceived of, implemented and become an integral part of some future health care system is an important and challenging task that requires innovative theoretical treatments. PMID- 14534634 TI - Rescuing the emergency--multiple expertise and IT in the emergency field. AB - OBJECTIVES: The rapid growth, both in technical and social approaches to the innovation phenomenon in health care suggests that profound changes are occurring. The focus of the project was to evaluate these changes through the case study of the French emergency field. METHODS: The consequences of information technology implementations in the Emergency Services are presented as models to investigate the importance of technical expertise, with both the involvement of all the stakeholders from the beginning of the design and the test of the new tool in real settings. RESULTS AND CONCLUSION: A new form of work resulted for the management of the crisis, which questioned the status of the actors and their actions, and the characteristics of the protocols of rescue. The outcome of this project was a new model built for the rescue of the emergency field which encompasses alternative forms of expertise. PMID- 14534635 TI - Big is beautiful: electronic patient records in large Norwegian hospitals 1980s 2001. AB - OBJECTIVES: This paper aims to describe and analyze the prolonged efforts - spanning close to two decades - of developing and using electronic patient records in the large, university-based hospitals in Norway. METHODS: This study belongs to an interpretative approach to the development and use of information systems. RESULTS: The increase in organizational, institutional, political and technological complexity has been seriously underestimated. This paper describes and analyses the prolonged efforts - spanning close to two decades - of developing and using EPRs in the large, university-based hospitals in Norway. The investments involved were considerable, implying that a crucial aspect of these efforts has been the way alliances have been forged with public institutions and agendas. CONCLUSIONS: The conditions for small-scale, bottom-up and evolutionary approaches never succeeded in constructing themselves as a viable alternative to the larger, more sweeping electronic patient record initiative, reiterating a more general tendency to privilege the more comprehensive and daring projects. PMID- 14534636 TI - Design and introduction of an electronic patient record: how to involve users? AB - OBJECTIVES: Two forms of participatory design (PD) - management-dominated and end users focused - are described in the design and introduction of an electronic patient record (EPR) for a mental health care centre. METHODS: Qualitative research methods were used to achieve a 'fine-grained' insight into the EPR development process and its impacts. These methods included interviews, document study and observation of project group meetings. RESULTS: Management-dominated PD is highly structured, led by managers and centred around rationalisation of work. End-user focused PD is more bottom-up oriented. Central in this approach are users' working patterns and user needs. In the case described, both forms actually lead to a poorly functioning EPR. It is explained why these results could be expected. CONCLUSION: Our findings suggest not to reject PD, but to find a more appropriate balance between the two forms of PD. PMID- 14534637 TI - Order creation and communication in healthcare. AB - OBJECTIVES: The aim of this paper is to examine the adequacy of the concept of Physician Order Entry (POE) as a model for clinical systems, and to suggest an alternative understanding of the order creation and communication process. METHODS: The study is based on an interpretative analysis of POE as a model for clinical systems and the results of our recent fieldwork. RESULTS: Observations from our recent fieldwork suggest that orders, like patient care in general, emerge from interactions among patients, physicians, nurses, family members, and others, employing a variety of technologies and information resources in the process. Orders as we have observed them originate, are negotiated, and are carried out in a dynamically evolving group with fluctuating membership and shifting role responsibilities. Furthermore, orders by themselves represent only a partial picture of what is done for the patient. CONCLUSION: We argue that information systems are more likely to be helpful if they accommodate and facilitate POE as a multidisciplinary collaboration effort and fit better into the larger system of patient care. PMID- 14534638 TI - Design as bootstrapping. On the evolution of ICT networks in health care. AB - OBJECTIVES: This paper assumes that in addressing major challenges related to telemedicine as networks enabling huge improvements of heath services we need to move beyond complexity and rather focus on the very nature of such networks. METHODS: The results of this paper are based on an interpretive analysis of three case studies involving telemedicine, i.e. broadband networks in minimal invasive surgery, EDI infrastructures and telemedicine in ambulances. RESULTS AND CONCLUSION: The well-known concept of "critical mass" focuses on the number of users as a significant factor of network growth. We argue however, that we should not only consider the size of the network, but also the heterogeneity of its elements. In order to discuss heterogeneity along several dimensions, we find Granovetter's and Schelling's models of diversity in individual preferences helpful. In addition to the heterogeneity of the individual users, we discuss heterogeneity related to use areas and situation, to technologies, etc. The interdependencies and possible conflicts between these dimensions are discussed, and we suggest "bootstrapping" as a concept to guide the navigation/exploitation in/of these dimensions. PMID- 14534639 TI - Working IT out in medical practice: IT systems design and development as co realisation. AB - OBJECTIVES: The paper explores possibilities for situating IT design and development work within the context of use so as to support the co-realisation of technology and 'design in use'. The aim is to build a new understanding between IT professionals and users which is grounded upon what happens as the latter grapple with the problems of applying IT, appropriating its functionalities and affordances into their work practices and relations. METHODS: Following a discussion of participatory design and ethnomethodology, a novel method called co realisation, which aims to provide a synthesis of the preceding methods, is suggested as an alternative. Through a discussion of findings from a case study of IT systems design and development in healthcare we show how the co-realisation approach might provide work-affording systems and how user-designer relations might be reformulated. We suggest that work-affording systems can be developed through the deployment of an engaged facilitator who works with the users to unpack the work site-specific potentialities of technology. RESULTS: The case study shows how risk of non-adoption might be minimised through the development of partnerships, and how the presence of the facilitator in the workplace capitalises on the mundane work undertaken therein and how the facilitator might work with the users to develop artefacts that support this work as opposed to reconfiguring it. CONCLUSIONS: The case study illustrates co-realisation in action and how it might be seen to reconfigure relations between users and designers in a way that appears productive. Co-realisation can help address the widely observed problem of IT systems failures in healthcare. PMID- 14534640 TI - An activity-theoretical method for studying user participation in IS design. AB - OBJECTIVES: This paper aims to present an activity-theoretical method for studying the effects of user participation in IS development. METHODS: This method is developed through a case study of the process of designing a diabetes database. RESULTS AND CONCLUSIONS: The method consists of a historical analysis of the design process, an ethnographical study of the use of the database, and researcher-driven interventions into the on-going user-producer interaction. In the historical analysis, we study particularly which user groups of the database have influenced the design work and which perspectives need to be incorporated into the design in the near future. An analytical model consisting of perspectives on local design, particular technology, and societal domain is introduced as a conceptual tool for this analysis. We also introduce the possibility of employing the historical analysis in guiding an ethnographical study of the user sites and researcher-driven interventions, which provide the participants with tools for improving their design process. PMID- 14534641 TI - The 'Seamless Web': the development of the electronic patient record in Aarhus region, Denmark. AB - OBJECTIVES: The article surveys the organization of the current project to develop an electronic patient record in the Aarhus Region, Denmark. METHODS: The article is based on various policy documents and reports as well as a number of semi-structured interviews with project managers from the EPR organization in Aarhus County and with participants in the development process at local hospitals. This material is used to present and discuss the framing of the project in a 'discourse coalition'. RESULTS: The stabilization of a specific discourse coalition has been an important factor in ensuring the success of the development project up to the present moment. This coalition became relatively stable by integrating a diverse set of actors in a story-line about the relationships between co-operation, management and technology in the medial sector, and has influenced the modular organization of the project. CONCLUSIONS: The successful maintenance of the discourse coalition allows the project to appear 'seamless' from the outside. Conversely, the project is likely to be continually reviewed as successful only to the extent that it is able to flexibly keep the fluctuating set of relevant actors in alignment. If the practical work of keeping a coalition in place remains invisible it becomes easy to imagine an ideal way of planning large socio-technical projects, like developing an ECR. But practical success is more likely to be achieved if one takes seriously the thorough intertwining of discursive, organizational and technical aspects of development projects. PMID- 14534642 TI - "Computers can land people on Mars, why can't they get them to work in a hospital?" Implementation of an Electronic Patient Record System in a UK Hospital. AB - OBJECTIVES: This paper aims to describe and interpret the implementation of a hospital information system in a large UK hospital. METHODS: The paper is based on a longitudinal case study over a three-year period in which a cross section of hospital staff involved with the information system were interviewed. RESULTS AND CONCLUSIONS: Ambitious government targets for the use of Information Technology in the UK National Health Service sit alongside a history of notable project failures. The decision by a UK hospital to install an advanced, integrated electronic patient record system therefore faced conflicting demands and expectations. This paper suggests that its simple categorisation as either a success or failure is problematic. Rather, the differing viewpoints that lead some clinicians to express "disappointment" with its performance, while others described its features as "tremendous" and managers suggested that the system had become "taken for granted" are explored. A number of broader phenomena relating to the organisational processes surrounding information systems implementation are also identified. PMID- 14534643 TI - Managers see the problems associated with coding clinical data as a technical issue whilst clinicians also see cultural barriers. AB - OBJECTIVE: In UK general practice, the coding of clinical data (Read Coding) is far from universal. This study set out to examine the barriers to recording structured information in computerised medical records; and to explore whether managers and clinicians had different perspectives in how these barriers should be overcome. METHOD: A qualitative study, using semi-structured interviews of general practitioners, primary care nurses and practice managers. The interviews were recorded verbatim, and then underwent thematic analysis; additional interviews were conducted until thematic saturation was achieved. RESULTS: For clinicians the recording of structured data within a consultation is not a neutral activity, they are highly aware of diagnostic uncertainty and sensitive to the potential impact of both a correct and incorrect diagnostic label on their relationship with their patient. Clinicians accept that data has to be coded if they are to demonstrate that appropriate evidence based care has been provided to populations; but alongside this they require free-text as a more powerful reminder of the individual human encounter. Managers felt that they could encourage clinicians to code data for re-use as part of population data or as quality target indicators rather than as an enabler of the next consultation. CONCLUSIONS: The primary care consultation is a complex social interaction, and coding of the medical diagnosis in itself imposes the bio-medical model, carries assumptions about certainty, and is perceived by clinicians to potentially jeopardise their relationships with their patient. Further research to elicit patients' views may help clarify the magnitude of this barrier. PMID- 14534644 TI - Two patient care information systems in the same hospital: beyond technical aspects. AB - OBJECTIVES: To compare two clinical workstations in one hospital with respect to technical, organizational, cultural and human factors. One clinical workstation was a GUI to the HIS. The other was an electronic patient record for stroke. METHODS: Data were collected by means of in-depth interviews with end-users of both clinical workstations. The interviews were audio taped and transcribed for analysis. RESULTS: End users assessed both clinical workstations as user friendly. Coordination between health care workers was perceived to be enhanced. However, in both situations poor communication between management, implementers and users resulted in uncertainty and skepticism about future perspectives. Further-more, it appeared that inpatient and outpatient settings needed clinical workstations with different requirements for an optimal fit between work practices and information system. CONCLUSIONS: Regardless of the domain and content of a workstation, it can support coordination between disciplines. The communication concerning the information technology strategy deserves much attention. Finally, the requirements for inpatient and outpatient workstations differ. PMID- 14534645 TI - Integrating health information systems: a critical appraisal. AB - OBJECTIVES: The aim of this paper is to critically assess some of the underlying assumptions behind these initiatives and analyze the dominant expressions, forms and mechanisms of integration. METHODS: This paper employs a discourse analysis to elicit the notion of integration of information systems and its consequences for electronic patient record systems. RESULTS AND CONCLUSIONS: An alternative strategy is outlined, encouraging a more decentralized, multi-vocal approach acknowledging the productive role of related and duplicated information and preserving the existing variety of information systems. PMID- 14534646 TI - Medicine and aviation: a review of the comparison. AB - OBJECTIVE: This paper aims to understand the nature of medical error in highly technological environments and argues that a comparison with aviation can blur its real understanding. METHODS: This study is a comparative study between the notion of error in health care and aviation based on the author's own ethnographic study in intensive care units and findings from the research literature on errors in aviation. RESULTS AND CONCLUSIONS: Failures in the use of medical technology are common. In attempts to understand the area of medical error, much attention has focused on how we can learn from aviation. This paper argues that such a comparison is not always useful, on the basis that (i) the type of work and technology is very different in the two domains; (ii) different issues are involved in training and procurement; and (iii) attitudes to error vary between the domains. Therefore, it is necessary to look closely at the subject of medical error and resolve those questions left unanswered by the lessons of aviation. PMID- 14534647 TI - Sociotechnical requirements analysis for clinical systems. AB - OBJECTIVE: We explore sociotechnical requirements by examining the use of a computerized patient record system in an intensive care unit of a U.S. hospital and present two sociotechnical requirements, awareness and coordination, embedded in the users' work. METHOD: The study is based on observation during seven months of the use of a computerized patient record system in a surgical intensive care unit. During that period semi-formal interviews, informal interviews were held. RESULTS AND CONCLUSIONS: A key step in the design of clinical systems is the development and analysis of requirements. However, traditional requirements analysis is based on a set of assumptions that break down in the highly collaborative, exception-filled clinical domain. Sociotechnical requirement analysis enabled the designers to gather a much richer description of the environment surrounding the computer system, highlighting awareness and coordination, embedded in the users' work. PMID- 14534648 TI - Patient reports as stories of clinical work: narrative and work in neuroradiology. AB - OBJECTIVE: This paper describes the connections between the radiology report and clinical work and considers the implications for computerisation. METHOD: A story representation is described that allows consideration of the radiology report as an active unit of narrative rather than a passive collection of data. This paper draws upon the results of a qualitative study of a neuroradiology department. RESULTS AND CONCLUSION: Radiology reports recount a patient condition but also represent and influence clinical work. PMID- 14534649 TI - Teleconsultation: rejected and emerging uses. AB - OBJECTIVE: The aim of this paper is to analyse telemedicine as a new means to improve health care accessibility. METHOD: A case study design was used to understand how medical specialists perceived, made sense of, and appropriated a teleconsultation system. RESULTS AND CONCLUSIONS: The technology was used neither in the manner nor to the extent anticipated by its designers. A fundamental modification to the traditional medical consultation process has emerged. Unable to be used as a substitute to the traditional medical consultation process, the teleconsultation system imposes a greater burden on the shoulders of participating physicians who, after a few trials, returned to their traditional mode of practice. PMID- 14534650 TI - Integrating quantitative and qualitative methods in patient care information system evaluation: guidance for the organizational decision maker. AB - OBJECTIVE: The aim of this paper is twofold. First, we describe two important dimensions of patient care information systems (PCIS) evaluation: the domain of evaluation and the different phases of the PCIS implementation. Second, we claim that, though Randomized Controlled Trials (RCTs) are often still seen as the standard approach, this type of design hardly generates relevant information for the organizational decision maker. METHOD: Interpretive study of evaluation literature. RESULTS AND CONCLUSIONS: The field of evaluation is scattered and the types of questions that can be asked and methods that can be used seem infinite and badly demarcated. Different stakeholders, moreover, often have different priorities in evaluating ICT. The most important reason for the lack of relevance of RCTs is that they are ill suited for investigating why and how a PCIS is being used, or not, and what the (often unplanned) effects and consequences are. Subsequently, our aim is to contribute to the discussion about the viability of qualitative versus quantitative methods in PCIS evaluation, by arguing for a specific integration of quantitative and qualitative research methods. The joint utilization of these methods, we claim, yields the richest results. PMID- 14534651 TI - Point of care technology: a sociotechnical approach to home health implementation. AB - OBJECTIVES: This paper describes an ongoing evaluation study of the design and implementation of Point of Care (POC) technology in a home health agency. METHODS: During the implementation of POC technology data about user acceptance were collected with the help of surveys and statistically analyzed. The survey also allowed for entering non-structured observations. A time series study was conducted to identify factors associated with technology change and its effect on professional competency. RESULTS AND CONCLUSIONS: Healthcare markets and consumers are demanding that providers use sophisticated information technology to document, transmit and access clinical and financial information across all delivery points. Newly adapted Patient Care Information Systems (PCIS) are available for clinical use in home care. The factors associated with home care nurses' end user acceptance and use of the POC technology in clinical work are described. A sociotechnical approach with selected change management strategies appears to have positively influenced the nurse end users' POC technology adoption and use in clinical work. PMID- 14534652 TI - Resistance to computerized care planning systems by qualified nurses working in the UK NHS. AB - OBJECTIVE: This study investigated whether computerized systems, designed to produce detailed plans for the nursing care of inpatients, were resisted by the nurses who were expected to use them. METHODS: Qualitative study, using semi structured interviews with nurses working in the UK National Health Service. RESULTS AND CONCLUSIONS: Resistance took the form of 'resistive compliance,' and this resistance is analyzed and explained. Resistance can best be understood in terms of the culture of nursing. This implies that the design and implementation of computerized systems in health care should take these factors into account. PMID- 14534653 TI - Positioning the patient: normative analysis of electronic patient records. AB - OBJECTIVE: This paper aims to take the current ethical research on electronic patient records beyond the ethical-legal issues of privacy to include contemporary issues of the 'politics of technology' and 'value sensitive design'. METHODS: The paper employs an interpretive approach to analyze research on electronic patient records with concepts of the politics of technology and value sensitive design. RESULTS AND CONCLUSIONS: The broad development towards computerization of patient records lacks needed attention to the potential effects of such systems on the patients. This paper argues for a broader understanding of the normative aspects involved in the design and implementation of electronic patient records (EPRs). It suggests three supplemental dimensions for normative analysis of EPR design relevant to the position of the patient: presence, agency, and identity. These dimension are discussed and illustrated as concepts connecting ethical dimensions of patienthood with design features of EPRs. PMID- 14534654 TI - Measuring the completeness and currency of codified clinical information. AB - OBJECTIVES: The paper describes how an objective score (CCscore) of the 'completeness' and 'currency' of codified clinical information relevant to the management of diabetes mellitus may be derived for individual practices. METHODS: A questionnaire was developed and administered to 35 practices and statistical methods were used to test for correlation between the prevalence for diabetes mellitus and the relevant CCscores RESULTS: No significant correlation could be found. CONCLUSIONS: The 'quality' of computer-stored information varies widely across English General practices for reasons that are incompletely understood. We demonstrated how CCscores may be calibrated for different 'views' of 'relevance', 'completeness', and 'currency' and yet be consistent across practices for a given 'view'. The potential value of this score and how it may contribute to our understanding of variation in 'information quality' are discussed. PMID- 14534655 TI - "We fill in our working understanding": on codes, classifications and the production of accurate data. AB - OBJECTIVES: This paper describes differences in the way general practitioners in Denmark, The Netherlands and Great Britain make codes fit into the local conditions under which they work. METHODS: An ethnographic study method has been used to collect data in Dutch, British and Danish general practices. RESULTS AND CONCLUSIONS: The paper argues that what counts as "accurate data" is locally constructed. As codes are produced in local networks of human and technological actors, the way accuracy is constructed is dependent on the extra work that is carried out (by actors inside the clinic as well as outside of it). On the basis of differences between coding practices and classification systems the paper discusses how inherent tensions between coding for primary and secondary purposes can be solved. The paper concludes that instead of evaluating data in terms of how accurate they are in general, they should be looked at in terms of pertinence to specific research questions. PMID- 14534656 TI - Portal fibrosis and schistosomal portal hypertension: what is the best strategy for primary and secondary prevention of hemorrhage from esophageal varices. PMID- 14534657 TI - The influence of periportal (pipestem) fibrosis on long term results of surgical treatment for schistosomotic portal hypertension. AB - AIM: To evaluate the degree of influence that periportal fibrosis has on clinical development and the long term results of surgical treatment on patients with hepatic-splenic schistosomiasis with previous gastrointestinal hemorrhages. METHODS: During the period of 1992-1998, 111 patients underwent surgical treatment for the treatment of hepatic-splenic schistosomiasis with previous gastrointestinal hemorrhages. The degree of fibrosis was classified as: degree I the portal spaces show a rich increase of young connective cells, a slight collagen production and a varying presence of inflammatory infiltrate. The periportal blade unchangeable (29/111); degree II - there is an expansion of the connective tissue with the emission of radial collagen septa, producing a star shaped aspect (38/111); degree III - the connective septa form bridges with other portal spaces or with the vein, with evident angiomatoid neo-formation (44/111). CONCLUSION: The patients with periportal fibrosis degree I present recurrent hemorrhages statistically less than patients with periportal fibrosis degrees II and III, and that the intensity of the periportal fibrosis is not the only pathophysiological factor of the esophageal varices, gastric varices, prevalence of post-operative portal vein thrombosis and hematological and biochemical alterations of the patients with pure mansoni schistosomiasis. PMID- 14534658 TI - [Bacterial infection in cirrhotic patient]. AB - BACKGROUND: Bacterial infections at admission or during hospitalization are frequent complication of cirrhosis that occurs in about 30% of the cases. Furthermore they are responsible for 25% of deaths in this population. AIM: Evaluate the prevalence of bacterial infections in cirrhotic patients at a general hospital and determine its correlation with alcoholic etiology of liver disease; degree of hepatic dysfunction and upper gastrointestinal bleeding. PATIENTS/METHODS: Five hundred and forty one admissions were retrospectively evaluated in 426 cirrhotic patients at years 1992 to 2000. The mean age was 50.5 years (15-95), being 71.2% male. The alcoholic etiology of cirrhosis was 35.4%. The main outcome considered was discharge or death during admission. RESULTS: One hundred and thirty five episodes of bacterial infections (25%) were diagnosed. The most frequent are urinary tract infection (31.1%), spontaneous bacterial peritonitis (25.9%) and pneumonia (25.2%). The association between urinary tract infection and pneumonia occurred in 3.7% and erysipelas or cellulites in 11.1%. Bacteremia occurred in 2.9%. There was a correlation between bacterial infection and alcoholic etiology of liver disease, hepatic dysfunction and upper gastrointestinal bleeding. The mortality was higher in the infected patients (8.9%) and in those with a poor hepatic function. CONCLUSIONS: Bacterial infections are common complications in cirrhotic patients and are correlated with alcoholic etiology, Child Pugh classification and upper gastrointestinal bleeding. Furthermore, bacterial infections are correlated with poor prognosis. PMID- 14534659 TI - [Megaesophagus microbiota and carcinogenesis]. AB - BACKGROUND: The risk of development of spin cell carcinoma of the esophagus is 33 times higher in patients with chagasic achalasia. It is possible that the production of N-nitroso compounds in the esophageal lumen by of bacterial action in the stasis liquid that reduce nitrates from diet into nitrites may play a role in this process. AIM: To analyze qualitatively and quantitatively the microbiota in chagasic megaesophagus with special attention to bacteria capable of transforming nitto reduction. PATIENTS: Fifteen patients (six men and nine women) were prospectively studied, with ages varying from 28 to 73 years. Patients were divided into three sub-groups according to Rezende et al. classification of esophageal dilation (grade I, grade II and grade III). METHOD: The sample collection was performed using a method specially developed to avoid contamination with microorganisms of the oral cavity and oropharynx, using a Levine catheter n 14 and a 7,5 oro-traqueal tube. RESULTS: Ninety three point three percent of the cultures were positive, with great bacterial variability and predominance of a variety of aerobic Gram-positive and anaerobic bacteria. The bacterial concentrations were generally more elevated in grade III in comparison to grade I and grade II. Among the microorganisms found, Staphylococcus sp, Corynebacterium sp, Peptostreptococcus sp e a Veillonella sp were those with the capability of nitrate reduction. CONCLUSION: It was concluded that patients with megaesophagus present some bacteria in the esophageal lumen that are able to reduce nitrates intro nitrites, an important step in the formation of N-nitroso compounds. PMID- 14534660 TI - [Recommended diet for reflux spectrophotometry]. AB - BACKGROUND: The spectrophotometric probe, which uses bilirubin as a marker for the detection of duodenoesophagic reflux is subject to interference from strongly colored foods, which can cause erroneously high bilirubin absorbance readings. To overcome this problem it is necessary to ingest a diet that is free from such substances. OBJECTIVE: To test the absorbance of 48 different food substances in an in vitro environment. MATERIAL AND METHOD: Dry foods were blended with water or milk and non-dry solid foods were blended undiluted. It was utilized the proper calibration recipient to test them. RESULTS: The absorbance of weakly colored foods was usually lesser than the commonly accepted threshold of 0.14, and the absorbance of strongly colored foods was usually above this. Thirty-two from the 48 substances tested are suitable when the absorbance threshold is set at 0.14, but scrambled eggs, lacteous flour mush, green beans, beetroot, carrot, chayote, squash, "baroa" potato, boiled corn, orange, cashew, purple grape, avocado, mango, papaya and peach can alter the results and must be avoided. CONCLUSION: From the foods evaluated, enough are suitable at the 0.14 threshold to enable a suitable diet to be constructed for most patients during Bilitec studies. PMID- 14534661 TI - [Leukocyte count, C reactive protein, alpha-1 acid glycoprotein and erythrocyte sedimentation rate in acute appendicitis]. AB - BACKGROUND: The diagnosis of acute appendicitis is clinic, but in some cases, it can present unusual symptoms. The diagnostic difficulties still lead surgeons to unnecessary laparotomies, which reach rates from 15% to 40%. Laboratory exams, then, may become important to complement appendicitis diagnosis. The leucocyte count seems to be the most important value, but measurement of acute phase proteins, specially, the C-reactive protein, is object of several studies. PATIENTS AND METHODS: This was a prospective study, involving 63 patients submitted to appendecectomies for acute appendicitis suspicion, in "Hospital das Clinicas", Federal University of Uberlandia, MG, Brazil, in whose blood were made dosages of acute phase proteins and the leucocyte count. RESULTS: The sample was composed by 44 male and 19 female patients, and the majority of them was between 11 and 30 years of age. The flegmonous type was the most freq ent (52.4%). The leucocyte count was altered in 74.6% of the cases and C-reactive protein elevation was observed in 88.9%. The alfa-1 acid glycoprotein and the erithrocyte sedimmentation rate were predominantly normal. The C-reactive protein was augmented in more than 80% of the cases in all ages. Leucocyte count and C reactive protein were altered in 80% of the patients with the limit of 24 hours from the beginning of symptoms. With clinical evolution time superior than 24 hours, the leucocyte count was altered in 69.7% of the cases, whereas C-reactive protein was in 97%. Sensibility and specificity of the leucocyte count were 88.7% and 20%. For the C-reactive protein, the values were, respectively, 88.9% and 10%. C-reactive protein dosage presented more sensible in cases with more than 24 hours of evolution (96.9%), although with no specificity. The alfa-1 acid glycoprotein and erithrocyte sedimmentation rate presented low sensitivity and specificity. CONCLUSION: The leucocyte count and the C-reactive protein present significantly altered in acute appendicitis cases, independent from genre or age interval. The leucocyte count and, mainly, the C-reactive protein must be considered in individuals with more than 24 hours of clinical evolution. Augmented values, as a matter of fact, should never substitute the doctor's clinical examination, but complement it. The erithrocyte sedimmentation rate and the alfa-1 acid glycoprotein do not contribute to acute appendicitis diagnosis. PMID- 14534662 TI - [Prevalence of heartburn in Pelotas, RS, Brasil: population-based study]. AB - OBJECTIVE: Estimation of the prevalence of heartburn in an adult population of Pelotas, RS, Brazil, once there is no national study done with adequate and specific statistic analysis. DESIGN: Cross-sectional study in populational base. METHOD: Were included citizen over 20 years in samples obtained in 40 populational areas according to Brazilian Institute of Geography and Statistics. An uniform and precodified questionnaire was applied to all participants. A quality control was done using a simplified questionnaire in 10% of the people that were included in the sample. Codification and the input of the data was done two times through the Epi-Info programme in order to minimize the consistency errors. The analysis of the figures was done in the SPSS programme and obtained comparations between the prevalence of heartburn in relation to demographic variables through the prevalence rate, confidence interval of 95% and the 'x - square test. Also were analysed the figures of heartburn associated to meals and decubit position. RESULTS: Among 1,263 persons included in the study were found rates of the heartburn prevalence of 48.2%, 32.2% and 18.2% in the last year, month and week, respectively. All forms of heartburn were associated to females with statistic significance. There was no association with age. The prevalence of heartburn associated with meals and decubit position in the last year was 33% and 16.6%, respectively. These association in the last month was 23% and 11.4% and in the last week was 14.3% and 7.2%, respectively. CONCLUSIONS: The high prevalence found in this study should call attention to those persons which could be exposed to the risk of complications, as well as to those that have alterations in the life quality. PMID- 14534663 TI - Stapled hemorrhoidectomy for the treatment of hemorrhoids. AB - BACKGROUND: The use of circular staplers in the treatment of hemorrhoidal disease is known as a simple procedure, with low morbidity, less post-treatment pain and with the same efficacy when compared to the classical hemorrhoidectomy. AIM: Analyze the operative technique, intra-operative and immediate postoperative complications and late results in 100 patients treated for hemorrhoid disease by stapling technique. PATIENTS AND METHODS: The group included 53 males and 47 females with mean age of 49.8 years, operated during the period June 2000 to June 2002 in the "Hospital Universitario" (Sao Paulo University Hospital) and "Hospital Sirio Libanes", in Sao Paulo, SP, Brazil. RESULTS: The majority of patients (78%) were discharged on the first post-operative day. Eight patients required supplementary analgesia and were given intramuscular diclofenac sodium and four of them received intramuscular tramadol. One intraoperative complication was bleeding which was difficult to control and required a blood transfusion. One patient was reoperated on the first postoperative day due to intermittent and persistent bleeding, however without hemodynamic changes or a drop in hematocrit. Two patients presented hemorrhoidal thrombosis in the early postoperative stage. The postoperative follow-up displayed: recurrence of prolapse, five cases (5%); anal sub-stenosis, two cases (2%); anal fissure, one case (1%); persistent pain, two cases (2%). Seven reoperations were performed: one due to bleeding, one due to sub-stenosis and five due to recurrence of hemorrhoidal prolapse and persistence of symptoms. CONCLUSIONS: Stapling is simple to accomplish, has low postoperative pain and rate of complications, however, the incidence of late reoperations is rather high and therefore major follow-up for better analysis is required. PMID- 14534664 TI - Effect of antihypertensive agents on stellate cells during liver regeneration in rats. AB - BACKGROUND: Although most studies have focused on the hepatocytes, all the hepatic cells participate in the regenerative process, among them the stellate cells. The stellate cells are mesenchymal cells involved in local neurotransmission and paracrine regulation of several liver functions. Acute hepatic tissue loss promotes the proliferation and activation of stellate cells from a quiescent state to myofibroblast-like cells. AIM: Investigate the effects of antihypertensive agents on the stellate cell population during the liver regenerative phenomenon in rats. METHODS: Adult male Wistar rats received lisinopril, losartan, bradykinin, or saline solution in a proportional volume, intraperitoneally, before and after 70% partial hepatectomy. Animals from the experimental and saline groups were sacrificed at 36 hours after partial hepatectomy. The alpha-smooth muscle actin labelled stellate cells population was counted in the periportal and pericentral zones of the liver specimen. RESULTS: The labelled stellate cells were more numerous in the control group both in the periportal and pericentral zones at 36 hours after partial hepatectomy than at the other times. The population of stellate cells was significantly lower in the losartan group and higher in the bradykinin and lisinopril groups than in the control group. CONCLUSIONS: These results suggest that losartan can inhibit and bradykinin and lisinopril can stimulate the stellate cell population during liver regeneration in rats. These cells synthesize several substances to stimulate liver regeneration. PMID- 14534665 TI - [Laparoscopic treatment of peri-appendicitis caused by intrauterine contraceptive device]. AB - BACKGROUND: Intrauterine device may perforate the uterus and cause several complications. AIM: To report a case of laparoscopic treatment of peri appendicitis caused by intrauterine device. PATIENT AND METHOD: A young female presented with pain in the right lower abdomen of 4 days of duration. On the physical examination, she had localized abdominal pain in the right lower abdomen with discrete guarding and pain on percussion and rebound tenderness. Ultrasonography showed an intrauterine device outside of the uterus, in the right lower of the abdomen. RESULTS: At laparoscopy, an inflammatory mass that consisted of the intrauterine device blocked by the mesoappendix and the appendix was observed. Appendectomy and removal of the intrauterine device were performed. CONCLUSION: Laparoscopy allows adequate treatment of peri-appendicitis caused by intrauterine device, inclusive with complete evaluation of the entire pelvic region to rule out associated complications PMID- 14534666 TI - [Overwhelming postsplenectomy infection]. AB - BACKGROUND: Splenectomy performed at any age and for any reason increases the risk for death due to overwhelming infection. AIM: To evaluate definition, etiology, incidence, risk factors and prophylaxis of overwhelming postsplenectomy infection, as well as the methods related to splenic tissue preservation when total splenectomy is necessary. METHOD: Bibliographic review. RESULTS: The etiological agents more frequently found are Streptococcus pneumoniae, Haemophilus influenza and type B, and Neisseria meningitidis. Other bacteria like Escherichia coli, Streptococcus b-hemolytic, Staphylococcus aureus and Pseudomonas sp represent a significant risk as well. In addition, a great variety of agents including other enteric Gram-negative microorganisms and non-bacteria pathogens can also be sporadically identified. The prophylaxis is based on three main aspects: patient's education, immune prophylaxis and chemical prophylaxis. However these are not enough to prevent the higher risk of developing sepsis. When total splenectomy is unavoidable, heterotopic splenic autotransplantation seems to be the only alternative for splenic tissue preservation. According to clinical and experimental studies, the splenic autotransplanted tissue present a similar structure to a normal spleen and preserve the splenic immune function. CONCLUSIONS: The high risk of overwhelming postsplenectomy infection, reduced the indication for total splenectomy, in trauma and several diseases. Prophylactic methods have been developed to minimize the effects of the sepsis. Several researches have been done to determine the immunocompetence of autogenous splenic grafts in response to bacteria invasion. PMID- 14534667 TI - The impact of Helicobacter pylori resistance on the efficacy of a short course pantoprazole based triple therapy. AB - BACKGROUND: Many of the currently used Helicobacter pylori eradication regimens fail to cure the infection due to either antimicrobial resistance or poor patient compliance. Those patients will remain at risk of developing potentially severe complications of peptic ulcer disease. AIM: We studied the impact of the antimicrobial resistance on the efficacy of a short course pantoprazole based triple therapy in a single-center pilot study. METHODS: Forty previously untreated adult patients (age range 20 to 75 years, 14 males) infected with Helicobacter pylori and with inactive or healing duodenal ulcer disease were assigned in this open cohort study to 1 week twice daily treatment with pantoprazole 40 mg, plus clarithromycin 250 mg and metronidazole 400 mg. Helicobacter pylori was assessed at entry and 50 3 days after the end of treatment by rapid urease test, culture and histology of gastric biopsies. The criteria for eradication was a negative result in the tests. Susceptibility of Helicobacter pylori to clarithromycin and metronidazole was determined before treatment with the disk diffusion test. RESULTS: One week treatment and follow up were complete in all patients. Eradication of Helicobacter pylori was achieved in 35/40 patients (87.5%) and was higher in patients with nitroimidazole-susceptible strains [susceptible: 20/20 (100%), resistant: 10/15 (67%)]. There were six (15%) mild adverse events reports. CONCLUSIONS: A short course of pantoprazole-based triple therapy is well tolerated and effective in eradicating Helicobacter pylori. The baseline metronidazole resistance may be a significant limiting factor in treatment success. PMID- 14534670 TI - Color Doppler sonography with contrast in the differentiation of ovarian tumors. AB - The objective of this study was to differentiate benign ovarian tumors from malignant ones before surgery using color and pulsed Doppler sonography, and to compare results obtained before and after use of contrast medium, thereby verifying whether contrast results in an improvement in the diagnostic sensitivity. METHODS: Sixty two women (mean age 49.9 years) with ovarian tumors were studied, 45 with benign and 17 with malignant tumors. All women underwent a transvaginal color Doppler ultrasonographic exam. A study of the arterial vascular flow was made in all tumor areas, as well as an impedance evaluation of arterial vascular flow using the resistance index. RESULT: Localization of the vessels in the tumor revealed a greater proportion of malignant tumors with detectable internal vascular flows (64%) than benign tumors with such flows (22%). There was a considerable overlap of these findings. The use of contrast identified a greater number of vessels with confirmation in the totality of tumors, but did not improve the Doppler capacity in tumoral differentiation. Malignant tumors presented lower values of resistance index than the benign ones, whether or not contrast was used. The cutoff value for resistance index that better maximized the Doppler sensitivity and specificity was 0.55. Through this value, an increase of the sensitivity after contrast use was obtained, varying from 47% to 82%, while specificity remained statistically unchanged. CONCLUSION: Although the injection of a microbubble agent improved the sensitivity of the method detecting vascularization of tumors, a positive finding for vascularization by this method was not clinically useful in the differentiation of benign and malignant ovarian tumors. PMID- 14534671 TI - Short- and long-term outcomes of ileal pouch-anal anastomosis for ulcerative colitis. AB - Ileal pouch-anal anastomosis was an important advancement in the treatment of ulcerative colitis. The aim of this study was to determine whether early complications of ileal pouch-anal anastomosis in patients with ulcerative colitis are associated with poor late functional results. PATIENTS AND METHODS: Eighty patients were operated on from 1986 to 2000, 62 patients with ileostomy and 18 without. The early and late complications were recorded. Specific emphasis has been placed on the incidence of pouchitis with prolonged follow-up. RESULTS: The ileostomy was closed an average of 9.2 months after the first operation. Fourteen patients were excluded from the long-term evaluation; 6 patients were lost to regular follow-up, 4 died, and 4 patients still have the ileostomy. Of the 4 patients that died, 1 died from surgical complications. Early complications after operation (41) occurred in 34 patients (42.5%). Late complications (29) occurred in 25 patients as follows: 16 had pouchitis, 3 associated with stenosis and 1 with sexual dysfunction; 5 had stenosis; and there was 1 case each of incisional hernia, ileoanal fistula, hepatic cancer, and endometriosis. Pouchitis occurred in 6 patients (9.8%) 1 year after ileal pouch-anal anastomosis, 9 (14.8%) after 3 years, 13 (21.3%) after 5 years, and 16 (26.2%) after more than 6 years. The mean daily stool frequency was 12 before and 5.8 after operation. One pouch was removed because of fistulas that appeared 2 years later. CONCLUSIONS: Ileal pouch anal anastomosis is associated with a considerable number of early complications. There was no correlation between pouchitis and severe disease, operation with or without ileostomy, or early postoperative complications. The incidence of pouchitis was directly proportional to duration of time of follow-up. PMID- 14534672 TI - Use of licit and illicit drugs at the University of Alfenas. AB - This paper reports the study of drug consumption carried out within the population of undergraduate students from 2 colleges of Alfenas, in the state of Minas Gerais state. Both licit and illicit drugs were studied, including alcohol, tobacco, marijuana, cocaine, heroin, crack, inhalants, glue, tranquilizers, stimulants, and others. METHODOLOGY: The research included a wide bibliographical search and the application of a questionnaire to approximately 23% of the students (total of 6500 students). RESULTS: A total of 1500 students participated in this investigation. The results demonstrated that there was a significant consumption of both licit and illicit drugs. The pattern of drug consumption in the research sample was similar to other investigations conducted in Brazil and in other countries. DISCUSSION: It was observed that 55% of the university students use drugs. However, the most surprising finding was that most of the students (88%) answered "yes" to the inquiry, "Have you already tried any type of drug, including alcohol and cigarettes?" The students revealed that they had taken drugs even prior to the admission to the university. The results suggest clearly that the university environment does not necessarily represent the starting point for student drug consumption. PMID- 14534673 TI - Clinical and epidemiological features of AIDS/tuberculosis comorbidity. AB - Considering the relevance of AIDS/tuberculosis comorbidity worldwide, especially in Brazil, this study was developed to describe the clinical and epidemiological features of the comorbid cases identified from 1989 to 1997 by the epidemiology service of the Hospital das Clinicas of the Universidade de Sao Paulo. METHODS: Databases containing information on all identified AIDS/tuberculosis cases cared for at the hospital were used to gather information on comorbid cases. RESULTS: During the period, 559 patients were identified as presenting with AIDS/tuberculosis comorbidity. Risk behavior for AIDS was primarily heterosexual contact (38.9%), followed by intravenous drug use (29.3%) and homosexual/bisexual contact (23.2%). Regarding clinical features, there were higher rates of extrapulmonary tuberculosis when compared to tuberculosis without comorbidity. There was an increase in reporting of AIDS by ambulatory units during the period. Epidemiologically, there was a decrease in the male/female ratio, a predominance in the 20 to 39 year-old age group, and a majority of individuals who had less than 8 years of schooling and had low professional qualifications. CONCLUSIONS: High rates of AIDS/tuberculosis cases at our hospital indicate the need for better attention towards early detection of tuberculosis, especially in its extrapulmonary form. Since the population that attends this hospital tends to be of a lower socioeconomic status, better management of AIDS and tuberculosis is required to increase the rates of treatment adherence and thus lower the social costs. PMID- 14534674 TI - Endovascular techniques for placement of long-term chemotherapy catheters. AB - PURPOSE: To analyze the results from using endovascular techniques to place long term chemotherapy catheters when advancing the catheter using the external jugular vein is difficult due to obstructions or kinking. METHODS: Between July 1997 and August 2000, 320 long-term chemotherapy catheters were placed, and in 220 cases the external jugular vein was used as the primary venous approach. In 18 of these patients, correct positioning was not achieved and several endovascular techniques were then utilized to overcome these obstacles, including manipulation of a J-wire with a moveable core, venography, and the exchange wire technique. RESULTS: In 94.5% of the patients with difficulties in obtaining the correct positioning, we were able to advance the long-term catheter to the desired position with the assistance of endovascular techniques. CONCLUSIONS: Venography and endovascular guidance techniques are useful for the placement of long-term catheters in the external jugular vein. PMID- 14534675 TI - Incisional hernia at the insertion site of the laparoscopic trocar: case report and review of the literature. AB - Laparoscopic operations offer a myriad of advantages resulting in a rapid postoperative recovery. Incisional hernia is an uncommon cause of morbidity in operative procedures performed by laparoscopic access, and the diagnosis may not be easily made. In our service we identified 2 patients with incisional hernia the site of trocar insertion. The records of these 2 cases were reviewed. We report on these 2 cases and present a review of the literature. PMID- 14534676 TI - Bile duct-duodenal fistula caused by AIDS/HIV-associated tuberculosis. AB - Although infrequent, digestive fistulae in HIV/AIDS patients have been reported throughout the digestive tract from the esophagus to the anus, with predominance of esophageal fistulae. AIDS/HIV-associated opportunistic infections may invade the digestive system and lead to fistula formation. Tuberculosis is the most common infection associated with these esophageal fistulae. We report here one case of bile duct-duodenal fistula in a female AIDS patient with associated abdominal Mycobacterium tuberculosis infection compromising lymph nodes of the hepatic pedicle where the fistula was found. According to the reviewed literature, this is the third case of bile duct-duodenal fistula associated with abdominal tuberculosis in AIDS patient, and the first where both the fistula and the tuberculosis infection were diagnosed at laparotomy for acute abdomen. Whether the AIDS patient with abdominal pain needs or not a laparotomy to treat an infectious disease is often a difficult matter for the surgeon to decide, as most of the times appropriate medical treatment will bring more benefit. PMID- 14534677 TI - The vacuum assisted closure of complex wounds: report of 3 cases. AB - Treatment of wounds using conventional methods is frequently limited by inadequate local wound conditions, or by a poor systemic clinical situation. Vacuum system may promote faster granulation tissue formation, remove excessive exudate, increase blood flow in the wound, and attract the borders of the wound to the center, reducing its dimension. We present 3 cases of patients with difficult wounds, due to bad local conditions, or poor clinical situation, in whom we used a vacuum system to prepare the wound for the surgical closure. One patient had a pressure ulcer, another had a diabetic foot ulcer, and the third one had an open foot stump. In the 3 cases a significant improvement of the wound conditions was achieved after 7 to 8 days, allowing successful surgical treatment with flap or skin grafts. PMID- 14534678 TI - Critical analysis of old and new vaccines against N. meningitidis serogroup C, considering the meningococcal disease epidemiology in Brazil. AB - Worldwide, the impact of meningococcal disease is substantial, and the potential for the introduction and spread of more virulent strains of N. meningitidis or strains with increased resistance to current antibiotics causes concern, making prevention essential. OBJECTIVES: Review the indications for meningococcal disease vaccines, considering the epidemiological status in Brazil. METHODS: A critical literature review on this issue using the Medline and Lilacs databases. RESULTS: In Brazil, MenB and MenC were the most important serogroups identified in the 1990s. Polysaccharide vaccines available against those serogroups can offer only limited protection for infants, the group at highest risk for meningococcal disease. Additionally, polysaccharide vaccines may induce a hypo responsive state to MenC. New meningococcal C conjugate vaccines could partially solve these problems, but it is unlikely that in the next few years a vaccine against MenB that can promote good protection against multiple strains of MenB responsible for endemic and epidemic diseases will become available. CONCLUSIONS: In order to make the best decision about recommendations on immunization practices, better quality surveillance data are required. In Brazil, MenC was responsible for about 2,000 cases per year during the last 10 years. New conjugate vaccines against MenC are very effective and immunogenic, and they should be recommended, especially for children less than 5 years old. Polysaccharide vaccines should be indicated only in epidemic situations and for high-risk groups. Until new vaccines against MenC and MenB are available for routine immunization programs, the most important measure for controlling meningococcal disease is early diagnosis of these infections in order to treat patients and to offer chemoprophylaxis to contacts. PMID- 14534679 TI - Cisplatin and platinum drugs at the molecular level. (Review). AB - Over twenty years of intensive work toward improvement of cisplatin, and with hundreds of platinum drugs tested, has resulted in the introduction of the widely used carboplatin and of oxaliplatin used only for a very narrow spectrum of cancers. A number of interesting platinum compounds including the orally administered platinum drug JM216, nedaplatin, the sterically hindered platinum(II) complex ZD0473, the trinuclear platinum complex BBR3464, and the liposomal forms Lipoplatin and SPI-77 are under clinical evaluation. This review summarizes the molecular mechanisms of platinum compounds for DNA damage, DNA repair and induction of apoptosis via activation or modulation of signaling pathways and explores the basis of platinum resistance. Cisplatin, carboplatin, oxaliplatin and most other platinum compounds induce damage to tumors via induction of apoptosis; this is mediated by activation of signal transduction leading to the death receptor mechanisms as well as mitochondrial pathways. Apoptosis is responsible for the characteristic nephrotoxicity, ototoxicity and most other toxicities of the drugs. The major limitation in the clinical applications of cisplatin has been the development of cisplatin resistance by tumors. Mechanisms explaining cisplatin resistance include the reduction in cisplatin accumulation inside cancer cells because of barriers across the cell membrane, the faster repair of cisplatin adducts, the modulation of apoptotic pathways in various cells, the upregulation in transcription factors, the loss of p53 and other protein functions and a higher concentration of glutathione and metallothioneins in some type of tumors. A number of experimental strategies to overcome cisplatin resistance are at the preclinical or clinical level such as introduction of the bax gene, inhibition of the JNK pathway, introduction of a functional p53 gene, treatment of tumors with aldose reductase inhibitors and others. Particularly important are combinations of platinum drug treatments with other drugs, radiation and the emerging gene therapy regimens. PMID- 14534680 TI - Intervention efficacy and malignant transformation to oral cancer among patients with leukoplakia (Review). AB - Elimination of leukoplakia is one of the strategies for prevention of oral cancer. However, the efficacy of reducing malignant transformation for the treated leukoplakia has rarely been reported despite a number of studies addressing its malignant transformation to oral cancer after intervention. The obstacle to do so is partially due to different lengths of follow-up and partially due to lack of information pertaining to the disease natural history that can be taken as a standard group for comparison. This work aimed to quantify the progression from leukoplakia to carcinoma with and without intervention after systematic literature review. The overall comparison of the efficacy of intervention across studies was therefore made. The literature between 1934 and 2001 was first reviewed, including two studies addressing the disease natural history model and 15 studies pertaining to intervention model in reducing malignant transformation. The simple constant-incidence exponential model and non constant incidence models (Weibull and Quadratic models) were therefore applied to estimate annual malignant transformation after intervention in various studies. Annual transition rates pertaining to the disease natural history were also estimated by Markov models. Intervention efficacy index using 10-year cumulative incidence from both models was also developed to assess intervention efficacy across studies. For the disease natural history, the estimates of annual transition rate of leukoplakia, annual transition rate from leukoplakia to oral cancer in the PCDP, and annual transition rate from the PCDP to clinical phase were 0.00121 (0.00019-0.00150), 0.0605 (0.0436-0.0755), and 1.8797 (0.13242 2.4352), respectively. Similar findings were observed in another retrospective study. Annual malignant transformation rates after intervention range from 0.0003 to 0.062 assuming constant incidence. Studies assuming non-constant incidence show different patterns of increasing or decreasing risk with time. The estimates regarding the efficacy of intervention for each study with different follow-up periods ranged from 42.9 to 99.3%. The heterogeneity of intervention efficacy due to a wide range of annual rates of malignant transformation was demonstrated in this review. This implies that prevention program for reducing malignant transformation may vary with different areas. Factors need to be considered including different diagnosis criteria, different histological type, distributions of risk factor, different patient resources, and different compliance rates or insufficient medical intervention. PMID- 14534682 TI - The usefulness of 99mTc-tetrofosmin SPECT in monitoring the response to chemotherapy in lung cancer patients. AB - We evaluated the usefulness of 99mTc-tetrofosmin (TF) scintigraphy in monitoring chemotherapy response in 31 inoperable lung cancer (LC) patients, 24 NSCLC and 7 SCLC. In all cases after 740 MBq 99mTc-TF i.v. injection both planar and SPECT images were acquired, before and after 3 cycles of chemotherapy; SPECT was analysed both qualitatively and semiquantitatively by calculating tumor/background ratio (T/B). Scintigraphic data were always related to CT findings, according to which patients were classified into 2 groups after therapy: responders (with >or=50% reduction in tumor size) and non-responders (with an increase or no change/no significant reduction in tumor size). Four patients were rechecked for the third time, during long-term follow-up. SPECT images, positive before therapy in all cases, were concordant with CT in assessing treatment response in 13/13 responders and in 18/18 non-responder patients, showing tumor reduction in the former and an increase or no change/no significant reduction in the latter. Planar imaging failed to give additional information but also led to disease down-staging in some cases. T/B ratio significantly decreased after therapy (1.67+/-0.39 vs. 3.02+/-0.87, p<0.005) in responders or=15 for Dukes' B patients (p=0.0388); (v-) vs. (v+) (p=0.0059), lymphatic invasion (ly-) vs. (ly+) (p=0.0435) for Dukes' A and B patients combined; D>n vs. D=n (p=0.0033), depth of tumor invasion or=se/a2 (p=0.0329) for Dukes' C patients. When the detection of >or=3 ONCs was defined as positive, the sensitivity, specificity, PPV, and NPV were respectively 77%, 100%, 100% and 71% in Dukes' B patients, as well as 75%, 72%, 73% and 74% in Dukes' C patients. The high-risk groups for recurrence/metastasis were identified by the following criteria: (v+) and (ly+), or=se/a2, and ONCs (+) of those with >or=2 factors for Dukes' C patients (selection rate; approximately 21.2-37.5%). These factors seem to be appropriate for separating patients into high-risk and low-risk groups of colorectal cancer recurrence/metastasis. PMID- 14534689 TI - Proliferative, infiltrative, and metastatic activities in colorectal tumors assessed by MIB-1 antibody. AB - MIB-1 antibody staining discriminates the cells in phases other than G0 of the cell cycle. The current study examined the proliferative activity assessed by MIB 1 antibody in colorectal adenoma, primary lesions of colorectal carcinoma (CRC) to investigate the relation between the histologic atypia, the proliferative, infiltrative, and metastatic activities. The MIB-1 antibody positive rate was immunohistologically determined in primary lesions in 311 patients, 22 having adenoma or carcinoma in situ, 207 invasive CRC without distant metastasis, and 82 invasive CRC with distant metastasis. The MIB-1 antibody positive rate was significantly higher in cases of adenoma with severe atypia and carcinoma in situ, showing a close relation between histologic atypia and proliferative activity. Among invasive CRC, the positive rate in poorly differentiated adenocarcinoma and mucinous carcinoma is significantly lower than in well differentiated and moderately differentiated adenocarcinomas. The positive rate was significantly lower in carcinomas with subserosa or deeper invasion than in carcinomas with submucosa or muscularis propria invasion, showing no distinct relation between the proliferative activity and the infiltrative activity. The positive rate of primary lesion was significantly lower in cases with metachronous liver or lung metastasis than in synchronous cases, indicating that metachronous hematogenous metastasis occurs even in cancers with low proliferative activity. The MIB-1 antibody positive rate showed a close relation between histologic atypia and proliferative activity in mucosal colorectal tumors although its relation with infiltrative activity was unclear in invasive CRC. It was apparent that metachronous hematogenous cancer metastasis might take place even in cases with low proliferative activity. PMID- 14534692 TI - Alpha-2,6-sialylation of L-PHA reactive oligosaccharides and expression of N acetylglucosaminyltransferase V in human diffuse large B cell lymphoma. AB - Cell surface sialylation and beta1-6 branching of L-PHA reactive oligosaccharides play an important role in metastatic capacities of various tumor cell lines. We analyzed the expression and sialylation of L-PHA reactive oligosaccharides in human diffuse large B cell lymphoma (DLBCL). DLBCL was grouped into three types; i). Group A, non-reactive type with no expression of L-PHA reactive oligosaccharides, ii). Group B, sialylated type with expression of sialylated L PHA reactive oligosaccharides and iii). Group C, non-sialylated type with expression of non-sialylated L-PHA reactive oligosaccharides. To clarify the linkage of sialic acid residues in L-PHA reactive oligosaccharides of Group B cases, L-PHA lectin histochemistry after treatment with two different neuraminidases was performed. In all Group B cases, L-PHA binding reactivity was found after treatment with Vibrio cholerae neuraminidase. But not after treatment with Newcastle disease virus neuraminidase. These data indicate that alpha2,6 linked sialic acid residues were predominantly involved in sialylation of L-PHA reactive oligosaccharides of Group B. To clarify the relationship between expression of N-acetylglucosaminyltransferase V (GnT-V), which catalyzes beta1-6 branching of L-PHA reactive oligosaccharides, and L-PHA reactivities in DLBCL, we investigated the expression of GnT-V using immunohistochemical methods. Most of the Group B and C cases expressed GnT-V while 33% of Group A cases showed no expression of GnT-V. These data suggest that expression of GnT-V is not always correlated with the expression of L-PHA reactive glycoconjugates. Furthermore, survival of patients in Group A which showed no expression of GnT-V was significantly shorter than that of patients in Group C which expressed GnT-V. Therefore, loss of non-sialylated L-PHA reactive oligosaccharides due to lack of expression of GnT-V in lymphoma cells may be associated with aggressiveness of DLBCL. PMID- 14534684 TI - Expression of the tumor suppressor gene PTEN is not altered in the progression of ovarian carcinomas and does not correlate with p27Kip1 expression. AB - This study was designed to investigate the role of PTEN in the progression of ovarian cancer. We performed mutation analysis and determined PTEN gene expression in tissue from both primary and relapsed cancers and in the corresponding occult metastases. Furthermore, p27Kip1 staining was conducted in order to explore a putative functional link. The study group comprised 112 tumor tissue specimens from 37 ovarian cancer patients. Expression of both PTEN and p27Kip1 was determined by immunohistochemistry. The PTEN mutational spectrum was determined by PCR-based sequence analysis. Fifty-six per cent of the tumors were positive for PTEN expression and 75% were p27Kip1 positive. For both markers, tumor cells ranged from 0 to 90% positivity. In 55% (20/37) of the cases, PTEN expression in the primary tumor was consistent and in the corresponding advanced cancer tissues, whereas the remainder showed considerable variation. p27Kip1 was consistently expressed in 16 out of 37 cases (43%). No mutations were observed in the coding region of the PTEN gene. No correlation was observed between PTEN and p27Kip1 expression. Our data indicate that expression of PTEN, but not p27Kip1 (one of the major mediators of PTEN function) is unchanged during the progression of ovarian cancer. This study suggests that in ovarian cancer PTEN does not play a major role in disease progression and is not involved in the alteration of p27Kip1 expression. PMID- 14534693 TI - Study of cancer gene therapy using IL-12-secreting endothelial progenitor cells in a rat solid tumor model. AB - To test the hypothesis that genetically modified bone marrow-derived endothelial progenitor cells (EPCs) can be effective carriers of therapeutic agents to tumor sites, we utilized our conditionally immortalized endothelial progenitor cell line, TR-BME-2. In the syngenic rat, systemically injected TR-BME-2 cells were immediately distributed to the organs (lung, bone marrow, peripheral blood, liver, spleen). Trapped cells were cleared within 4 days, but selective accumulation in the Walker256 tumor was maintained for over 4 days. The tumor growth was enhanced by administration of TR-BME-2 cells. It is suggested that accumulated TR-BME-2 differentiated to tumor vasculature, increased the tumor blood supply, and thereby increased the tumor volume. We conducted IL-12 gene transfection of TR-BME-2 cells with a virus vector in vitro, and used the resultant IL-12-secreting TR-BME-2 to deliver IL-12, which strongly activates cytotoxic lymphocytes and natural killer cells, to the tumor site in vivo. However, the tumor-progressive character of TR-BME-2 offset the anti-tumor effect of IL-12. Nevertheless, our results suggest that gene-transfected EPCs could be useful as a tumor-specific drug delivery system, especially if the tumor vasculature-promoting effect of EPCs can be blocked. PMID- 14534694 TI - Expression of cell cycle/growth regulator genes in human hepatocellular carcinoma and adjacent normal liver tissues. AB - We investigated the gene expression of the cell cycle/growth regulators in hepatocellular carcinoma (HCC) through the usage of Atlas human cancer array membranes, semi-quantitative reverse transcription polymerase chain reaction (RT PCR) and Northern blot. Hybridization of cDNA array membrane was performed with alpha-32P-labeled cDNA probes synthesized from RNA, isolated from HCC and adjacent non-cirrhotic normal liver. RT-PCR of 24 paired specimens and Northern blot of 4 paired specimens were used to confirm the expression patterns of the cell cycle/growth regulator genes identified by Atlas array hybridization. Among 79 genes related to cell cycle/growth regulators, transcription factor DP2 (TFDP 2) and E2F-3 were up-regulated, whereas dual-specificity mitogen-activated protein kinase kinase 1 (MAPKK1) and cell division protein kinase 3 (CDK3) were down-regulated in HCC. RT-PCR of TFDP-2 gave result consistent with Atlas human cancer cDNA array findings. Northern blot analysis of TFDP-2 and E2F-3 of 4 paired specimens all showed up-regulation in HCC compared to normal liver tissues. The results obtained from Atlas microarray provided for the first time a liver cancer-specific expression profile, which identified the gene expressions comprehensively and systematically. The findings may lead to better understanding of the mechanism of onset and progression of HCC. The rapid and high-throughout method of profiling gene expression by cDNA array provides an overview of the key factors that may be involved in HCC. Some genes are reported for the first time in HCC. PMID- 14534685 TI - Direct effects of octreotide, galanin and serotonin on human colon cancer cells. AB - The effects of mono, duple and triple treatment with octreotide, galanin and serotonin on a human colon cancer cell line (SW 620) were investigated. The cancer cells were exposed to a dose corresponding to 20 microg/kg body weight/day, and to 50 and 25% of this dose (0.2, 0.1 and 0.05 microg/ml). The cells were observed at the intervals: 3, 6, 12, 24 and 48 h. MTT-assay was used to determine numbers of viable cells. Proliferation and apoptosis were detected by immunocytochemistry using the avidin-biotin complex (ABC) method. The antibodies used were anti-Ki-67, anti-poly (ADP-ribose) polymerase 'PARP' and anti-Bcl-x. Proliferative and apoptotic indices were determined by computerized image analysis. Almost all the mono and duple treatments of the bioactive substances succeeded in reducing the numbers of viable cells. With triple treatment, however, this decrease was greater and was evident at all observation times. The effect on proliferation varied between none, and an enhancing or inhibiting action, depending on the dose, combination and observation time. The effect on apoptosis of mono or duple exposure to the bioactive gut substances varies, depending on the concentration, combination and observation time. Triple combination at the effective dose increased the apoptotic index, with the two markers used, and appeared after 6 h, extending up to 48 h. The reduction in the number of viable cancer cells was greater and occurred earlier than the increase in apoptosis and was observed whether the bioactive substances were used alone or in combinations and at different concentrations. It is therefore conceivable that some other mechanism(s) than apoptosis is involved which inhibits cancer cell respiration. It is possible that some of the dramatic in vivo changes seen earlier, following triple treatment with octreotide, galanin and serotonin, may have been direct effects of these substances on cancer cells. PMID- 14534695 TI - Heat shock protein 105 is overexpressed in a variety of human tumors. AB - We earlier reported that heat shock protein 105 (hsp105), which we identified by serological analyses of recombinant cDNA expression libraries (SEREX), was overexpressed in human colon and pancreatic adenocarcinoma. We went on to examine hsp105 expression in various colorectal cancers and adenomas, using immunohistochemical analysis. The 44 of 53 patients with colorectal cancers (83.0%) and only 2 of 21 (9.5%) with colorectal adenomas had an evident overexpression of hsp105, which means that overexpression of hsp105 is a late event in the colorectal adenoma-carcinoma sequence. Subsequently, we asked if hsp105 was overexpressed in other human tumors. During immunohistochemical studies, we discovered that overexpression of hsp105 occurred not only in cases of colorectal and pancreatic adenocarcinoma, but also thyroid, esophageal, breast, and bladder carcinoma and islet cell tumor, gastric malignant lymphoma, pheochromocytoma, and seminoma. On the other hand, hsp105 was evidently overexpressed only in the testis in human adult normal tissues. Thus, hsp105 is a useful marker of a variety of human tumors and hsp105 may prove to be a target molecule for designing anti-tumor immunotherapy. PMID- 14534696 TI - Maspin in thyroid cancer: its relationship with p53 and clinical outcome. AB - The serine protease inhibitor maspin has been reported to inhibit invasiveness and motility of tumor cells. Additionally, a p53-dependent regulatory pathway of maspin in human cancer has been indicated. In a pre-study we were able to detect maspin protein in papillary thyroid carcinomas (PTC), whereas normal (tumor-free) thyroid tissue, follicular adenomas, follicular carcinomas, poorly differentiated carcinomas and undifferentiated carcinomas of the thyroid were maspin-negative. The first aim of our study was to determine the prognostic value of maspin protein expression for the recurrence-free and overall survival of PTC patients undergoing radical thyroidectomy and postoperative irradiation. Secondly, maspin expression was correlated to p53 protein expression in order to gain additional information on a possible regulatory influence of the wild-type p53 protein on maspin. An immunohistochemical approach study was performed on 68 tumor specimens. Maspin protein expression was detectable in 48 of 68 patients (71%; M+). After a median follow-up of 81 (26-117) months the median recurrence-free survival was 60 (28-117) months for M+ and 42 (11-108) months for M- (p=0.03). After 110 months 83% of patients had recurrence-free disease in M+, whereas in M- only 40% of patients were recurrence-free. The median long-term survival was 81 (42-108) months for M+ and 55 (21-99) months for M- (p=0.03). After 5 years, M+ and M- patients had a total survival of 98 and 80%, and after 9 years 90 and 60%, respectively. Mutant-type p53 expression was detectable in 17 of 68 PTC (25%). Mt p53 was positive in 1 of 47 M+ (2%) compared with 16 of 20 M- (80%, p<0.01). This study indicates that maspin protein possibly functions as a clinically relevant inhibitor of tumor progression, preventing local invasiveness and further systemic progression of papillary thyroid carcinomas. Our data hint of a p53 dependent regulatory pathway of the maspin protein in human cancer. PMID- 14534698 TI - Intraperitoneal treatment using taxol is effective for experimental peritoneal carcinomatosis in a rat model. AB - Following resection of colorectal carcinoma, a considerable local recurrence rate within the previous tumor bed or at the peritoneal site remains an unsolved problem. Currently, there are no established protocols for the treatment or prevention of peritoneal carcinomatosis. Taxol showed benefit in patients with advanced tumor growth, in particular, gynecological carcinomas. Taxol was used to test whether it can either prevent or treat peritoneal tumor growth derived from colon carcinoma. In rats divided in 3 groups, peritoneal carcinomatosis was induced by tumor cell transfer: Taxol (170 mg/m(2)) was given i). directly (group A); ii). on days 5, 10, 15 postoperatively (representing early administration; group B), or iii). as late intraperitoneal (i.p.) chemotherapy (15, 20, 25 days following surgery; aiming for reduction of a manifest peritoneal carcinomatosis; group C) into the abdominal cavity. Tumor growth was quantified by tumor weight of the greater omentum and the mesenteric site, number of detectable tumor lesions, occurrence of hepatic and pulmonary metastases and the amount of ascites. Taxol was highly effective in preventing or reducing i.p. tumor spread when the drug was given directly or within a short time interval after tumor cell implantation (groups A and B), whereas no significant antineoplastic potential was found in the treatment of an established peritoneal carcinomatosis. In conclusion, Taxol appears to be a promising chemotherapeutic agent to be investigated in further detail with possible potential for a later human phase-I trial in peritoneal carcinomatosis. PMID- 14534697 TI - Complex chromosomal rearrangements in a secondary acute myeloblastic leukemia after chemotherapy in TRAPS. AB - We report on the cytogenetic findings from a patient with a de novo TNF-receptor associated periodic syndrome (TRAPS), who showed first symptoms at the age of four months. Thus, he obtained a long-term therapy with cortisone, chlorambucile, methotrexate and cyclophosphamide. At the age of 14 he developed a secondary acute myeloblastic leukemia. Highly complex chromosomal rearrangements were detected after banding analysis. The exact definition of karyotype and the involved breakpoints could only be resolved after application of sophisticated multicolor-FISH techniques: 44,XY,-5,der(6)t(6;7)(6pter right curved arrow 6q12::7p22.2 right curved arrow 7pter or 7pter right curved arrow 7p22.2), dic(7;19)t(6;19;6;7;19;7;19)(19qter right curved arrow 19q12::7p13 right curved arrow 7p11.1::19q12 right curved arrow 19p12 or 19p12 right curved arrow 19q12::7p11.1 right curved arrow 7q21.3::6q12 right curved arrow 6q26::19p13.3 right curved arrow 19p12::6q26-6qter),dic(12;13)(13qter right curved arrow 13p11.2::12p13.1 right curved arrow 12qter),ace(12;13)(13pter right curved arrow 13p11.2::12p13.1 right curved arrow 12pter), -19. The simultaneous presence of two dicentric chromosomes has not been reported previously and is striking, as such chromosomes are suggested to be instable. However, such chromosomes are observed frequently after chemo- or radiotherapy and in secondary, i.e. therapy related AML (tAML). Thus, AML in this case may result from a long-term therapy of TRAPS with methotrexate, cyclophosphamide, chlorambucile and cortisone. PMID- 14534699 TI - Cyclooxygenase-2 expression in lymph node metastasis of cervical and vulvar cancer. AB - Overexpression of cyclooxygenase-2 (COX-2), characterizes tumors with high potential for local invasion and lymph node involvement. We investigated the expression of COX-2 in primary tumors and metastatic regional lymph nodes (TDL) from untreated and chemotherapy treated cervical cancer, as well as vulvar cancer. Immunostaining of COX-2, expressed as values of COX-2 intensity density (COX-2 IDV) was performed on 57 metastatic TDL and 24 corresponding primary rumors from 14 cervical and 9 vulvar cancer patients admitted to the Department of Obstetrics and Gynecology, Catholic University of Rome. In 6 locally advanced cervical cancer tissue samples, from both primary tumor and TDL, were obtained after chemotherapy treatment. In untreated cervical cancer, COX-2 IDV in tumor cells from positive TDL were significantly lower (median 0.69, range 0.22-0.92) than those from primary tumors (median = 3.84, range 0.19-7.67) (p=0.011). In cervical cancer exposed to chemotherapy, COX-2 IDV in tumor cells from positive TDL were significantly lower (median = 2.06, range 1.48-6.52) than those from primary tumors (median = 6.4, range 4.5-13.7) (p=0.037). In vulvar cancer COX-2 IDV in tumor cells from positive TDL were lower (median = 0.39, range 0.02-6.09) than those from primary tumors (median = 2.49, range 0.71-8.10) (p=0.04). In conclusion, we showed that COX-2 expression is down-regulated in cervical and vulvar tumor cells invading the regional lymph nodes with respect to primary tumors, thus emphasizing the need for deeper insight into the tissue specific relation between tumor cells and node microenvironment. PMID- 14534700 TI - Pentoxifylline promotes development of murine colon adenocarcinoma-derived metastatic tumors in liver. AB - Pentoxifylline is commonly used in the treatment of peripheral vascular diseases. It improves microcirculatory flow and tissue perfusion. Moreover, pentoxifylline displays some immunomodulatory properties that presumably might affect the anticancer response. Therefore, the aim of the present study was to evaluate the influence of pentoxifylline on tumor development. Balb/c mice were injected with murine colon adenocarcinoma C-26 cells intravenously, into the vena portae, and divided into two groups. Mice from the experimental groups received daily intraperitoneal injections of pentoxifylline (30 mg/kg) while the controls were injected with 0.9% NaCl. Two weeks after C-26 cell inoculation mice were sacrificed and autopsy was performed. It was found that the livers of control animals revealed only several small tumor foci, whereas the livers of pentoxifylline-treated mice displayed numerous cancerous outgrowths. The mean liver weight in pentoxifylline group was 2.21+/-0.62 g as compared to 1.36+/-0.15 g in controls (P=0.004). Moreover, the influence of pentoxifylline on murine and human cell line proliferation in vitro was evaluated. It has been observed that pentoxifylline, at pharmacologically achievable concentrations, stimulated the proliferation of murine (C-26) and human (CaSki, U-937) cell lines. However, it did not stimulate human melanoma (WM-35) cell proliferation. Since there has been no evidence so far that pentoxifylline may promote tumor progression, it is still considered to be a safe drug. Moreover, some beneficial properties of pentoxifylline, which could be useful in cancer treatment, have been reported and a few clinical trials with oncological patients have been performed. Surprisingly, our study revealed that pentoxifylline significantly promoted C-26 derived metastatic tumor growth in liver. Although this model might be unique in its sensitivity to tumor-promoting effects of pentoxifylline, it cannot be excluded that similar effects might occur in some cases of tumors developing in humans. Such effects could be relevant, since the stimulatory influence of pentoxifylline on proliferation does not appear to be species- or tissue specific. PMID- 14534701 TI - Tumor-derived aberrant methylation in plasma of invasive ductal breast cancer patients: clinical implications. AB - Progressive p16 methylation has been associated with metastasis and invasive phenotypes in many cancers. Loss of E-cadherin (CDH1) function contributes to breast cancer progression by promoting cell proliferation, invasion and metastasis. Using methylation-specific PCR, aberrant hypermethylation of p16 and CDH1 in tumor and plasma was analyzed and correlated with levels of serum protein markers, carcinoembryonic antigen (CEA) and carcinoma antigen 15-3 (CA15.3), in 36 patients with invasive ductal breast cancer. Aberrant p16 methylation was found in 11% (4/36) of primary tumors and 8% (3/36) of plasma samples. Aberrant CDH1 methylation was detected in 25% (9/36) of primary tumors and 20% (7/36) of plasma samples. p16 and/or CDH1 hypermethylation was found in 31% (11/36) of primary breast carcinomas and 82% (9/11) of breast cancer patients with tumoral methylation showing identical epigenetic changes in plasma. The 25 patients without tumoral methylation did not show epigenetic changes in the plasma. Tumoral p16 methylation was significantly associated with advanced tumor stage (p=0.028; Fisher's exact test), tumor size (p=0.017) and nodal metastasis (p=0.002). However, p16 methylation in plasma was only associated with nodal metastasis (p=0.012). Altogether, aberrant p16 methylation in plasma and elevated serum CEA level were associated with advanced tumor stage (p=0.033), tumor size (p=0.022) and extensive nodal metastasis (p=0.003). With clinical implications, p16 hypermethylation in plasma and/or raised serum CEA levels may prove useful as diagnostic and prognostic markers for breast cancer. PMID- 14534702 TI - Paclitaxel and liposomal doxorubicin (Caelyx) combination in advanced breast cancer patients: a phase II study. AB - Due to the cumulative cardiotoxicity of doxorubicin in cancer treatment, the tendency of the physician to find a substitute has led to the use of liposomal encapsulated doxorubicin, as well as other similar compounds. Doxorubicin and paclitaxel, two of the most active agents for breast cancer, have often been used in combination for this condition. In the present study we combined liposomal doxorubicin with paclitaxel with the intention of diminishing the toxicity of the cardiac muscle. Twenty-three patients were evaluated for response rate, survival and toxicity. All patients had metastatic disease and were chemotherapy-naive after generalization of the disease. Liposomal doxorubicin was infused at a dose of 30 mg/m(2) and paclitaxel at a dose of 175 mg/m(2) once every 3 weeks. The response rate was complete in 2 patients (8.70%) and partial in 14 patients (60.87%) totalling 69.57%. The median duration of response was 6 months (range 2 13+) and median survival was 10 months (range 4-20+). Cardiotoxicity, myelotoxicity and gastrointestinal toxicity were well-tolerated. The high hand foot adverse reaction (47.83%) inhibited the continuation of treatment in half of the patients and due to this toxicity the trial was terminated after the 23rd patient. PMID- 14534703 TI - Repeated hepatic arterial infusion chemotherapy using an implanted port system in patients with unresectable malignant liver neoplasms: significant factors affecting early hepatic arterial occlusion. AB - The purpose of this study was to identify significant factors affecting early hepatic arterial occlusion in patients who received repeated hepatic arterial infusion chemotherapy using an implanted port system. Eighty-five patients with unresectable liver neoplasms who underwent implantation of the port system were studied. Arterial infusion chemotherapy was performed every 1-4 weeks. Arterial occlusion was evaluated by hepatic arteriography performed via the port every 3 months. Twenty variables were analyzed using univariate and multivariate analyses to identify significant factors affecting early hepatic arterial occlusion. Hepatic arterial occlusion was found in 25.9% (22/85) of the patients. Thirteen of them experienced early arterial occlusion within 6 months. The mean survival period was significantly worse in patients who experienced early arterial occlusion than those who did not (16 months vs. 26 months, p<0.05). In the multivariate analysis, the following 3 variables had independent value for early arterial occlusion; i). diameter of the common hepatic artery, ii). gender, and iii). previous systemic chemotherapy. Early arterial occlusion affects therapeutic effects and survival in patients who undergo arterial infusion chemotherapy with an implanted port. Factors demonstrated here are important to classify patients at risk of early hepatic arterial occlusion. PMID- 14534704 TI - Genetic polymorphisms of CYP1A1, GSTM1 and GSTT1 genes and lung cancer risk. AB - Genetic polymorphisms of the genes encoding for the xenobiotic metabolizing enzymes result in individual variations in the efficiency of detoxification of environmental carcinogens, and have been extensively associated with variable risk for lung neoplasms in different ethnic and environmental backgrounds. In this study, using PCR-RFLP based assays, we investigated the distribution of genetic polymorphisms in CYP1A1, GSTM1 and GSTT1 genes in Greek lung cancer patients (N=122) and healthy controls (N=178). The frequency of CYP1A1 m1 homozygous genotype was 0.04 in patients and 0.02 in controls (detected in 4.10% of patients and in 1.69% of controls, respectively), that of GSTM1 null genotype was 0.52 in patients and 0.54 in controls, whereas those of GSTT1 null genotype was 0.17 and 0.11, in patients and controls, respectively. The GSTM1 null genotype was more frequent in adenocarcinoma, as well as in lung cancer patients with history of chronic obstructive pulmonary disease (COPD). The GSTT1 null genotype correlated with advanced age of the patients at the time of diagnosis. Three combinations of rare genotypes - in subjects carrying simultaneously deviations from the common genotype in more than one gene - were over-represented in lung cancer patients, compared to control population, and were furthermore significantly associated with history of heavy tobacco consumption in lung cancer patients. The results imply involvement of specific genotype combinations of CYP1A1, GSTM1 and GSTT1 alleles in the development of lung cancer in heavy smokers. PMID- 14534705 TI - Somatic mutation in mitochondrial DNA and nuclear microsatellite instability in gastric cancer. AB - It was reported that somatic mutations in the mitochondrial DNA (mtDNA) are associated with high-frequency microsatellite instability (MSI-H) of the nuclear in gastric cancers. However, no correlation between mtDNA mutations and nuclear MSI-H was found in colorectal, breast, and renal cancers. Therefore, the association between mtDNA mutations and nuclear MSI-H in gastric cancers is controversial. We examined mtDNA mutations and nuclear MSI in a large panel of gastric cancers. One-hundred and five gastric cancers were selected. Mutations in the mononucleotide repeat (D310) of mtDNA and nuclear MSI at 5 microsatellite loci were examined by microsatellite assay. Somatic mutations in the mtDNA and nuclear MSI-H were detected in 16 (15%) and 14 (13%) of the gastric cancers, respectively. mtDNA mutations were detected in 2 of the 14 (14%) and 14 of the 91 (15%) tumors with and without nuclear MSI-H, respectively. There was no significant difference between them. These results suggest that somatic mutations in the mtDNA and nuclear MSI-H play important roles in gastric carcinogenesis, and that mtDNA mutations may not be associated with nuclear MSI-H in gastric cancers. PMID- 14534706 TI - Role of tumor-associated macrophages in renal cell carcinoma. AB - We investigated the biologic meaning of tumor-associated macrophages (TAM) in renal cell carcinoma (RCC). The study group comprised of 83 patients with RCC. TAM was isolated by plastic adherence following enzymatic digestion of surgically removed tumor tissues. In some of the patients, monocytes were also isolated from peripheral blood mononuclear cells by plastic adherence. When TAM and monocytes were compared in the same patients, TAM produced interleukin 6 (IL-6), tumor necrosis factor alpha (TNFalpha) and interleukin 1beta (IL-1beta) without lipopolysaccharide (LPS) stimulation, while monocytes hardly produced IL-6, TNFalpha and IL-1beta without LPS stimulation. However, with LPS stimulation, monocytes produced more IL-6, TNFalpha and IL-1beta than TAM. In stage T1 RCC patients, there was a significant positive correlation between TNFalpha production of TAM and tumor size. In order to investigate the effects of TAM on cancer cells, TAM was co-cultured with A498, K562 and in some cases, with short term established RCC lines for 96 h. As a result, TAM largely enhanced cell proliferation. These results suggested that TAM may play an important role in certain steps of tumor progression. PMID- 14534707 TI - Induction of tumor cell death by high hydrostatic pressure as a novel supporting technique in orthopedic surgery. AB - As vegetative forms of microorganisms are impaired by high hydrostatic pressure (HHP) in the range of 400-600 MPa, the non-thermal inactivation of vegetative bacteria, yeasts, and moulds present in foods such as jams, fruit juices, and dressings by HHP is now well-established. Eukaryotic cells, when subjected to HHP are also damaged. In the present study, the effect of HHP on cell viability of human osteoblasts (HOB), human fibroblasts (HFB), and different tumor cell lines such as osteosarcoma cells SAOS-2, human histiocytic leukemia cells U-937, and the ovarian cancer cell line OV-MZ-6 was investigated. Therefore the different cell lines were subjected to pressures between 50 and 400 MPa and tested for viability. At HHP of 100 MPa (10 min) about 80% of the various cell lines were still alive. At 350 MPa all of the cells were damaged and dead. The three tumor cell lines investigated were slightly more resistant to HHP (50% dead at 170-193 MPa) than HOB and HFB (50% dead at 130-145 MPa). The present study demonstrates that both normal cells and tumor cells are rapidly inactivated by HHP treatment. PMID- 14534708 TI - Alteration in the metastatic potential of ovarian cancer cells by transfection of the antisense gene of beta-1,4-galactosyltransferase. AB - beta-1,4-galactosyltransferase (beta-1,4-GT) has been reported to be activated in ovarian carcinoma cells and an isoform of this enzyme has been used as a tumor marker for ovarian cancer. The present study was undertaken to clarify how beta 1,4-GT affected the cell biological characteristics of ovarian cancer. To this end, we transfected an ovarian tumor cell line with an antisense gene of beta-1,4 GT. Proliferative potential and morphology of the cells transfected with the antisense gene did not differ from those of the control cells. Adhesive potential to the constituents of extracellular matrix was reduced in the antisense gene transfectants. In a nude mouse, the number of peritoneal dissemination foci of the antisense transfectants was smaller than that of the control cells. These results indicated that beta-1,4-GT is closely related to the invasive and metastatic potentials of ovarian cancer while it is not involved in the proliferative potential. PMID- 14534709 TI - Amphoterin induction in prostatic stromal cells by androgen deprivation is associated with metastatic prostate cancer. AB - Amphoterin, the major product of the high mobility group-1 gene, is a ligand associated with cancer invasion and metastasis through activation of the receptor for advanced glycation end products (RAGE). Expression of amphoterin and RAGE was examined in prostatectomy specimens from 40 patients with pT3 prostate cancer (18 non-metastatic and 22 metastatic) preoperatively treated with lutenizing hormone releasing hormone (LH-RH) agonist. Amphoterin expression was detected in tumor cells of 6 (27%) metastatic and 0 non-metastatic cases (p<0.0001). Amphoterin was also detected in prostatic stromal cells of 14 (63%) metastatic cases and 2 (11%) non-metastatic cases (p=0.0010). RAGE production was detected in cancer cells of 16 (73%) metastatic and 6 (33%) non-metastatic cases (p=0.0244). A total of 2 (22%) non-metastatic and 16 (73%) metastatic cases showed co-expression of amphoterin and RAGE in tumor cells or in tumor cells and stromal cells (p=0.0001). The in vitro invasive capacity of PC-3, a prostatic cancer cell line that co-expressed amphoterin and RAGE, was suppressed by treatment with amphoterin antisense S-oligodeoxynucleotide (ODN). Primary cultured human prostatic stromal cells secreted no amphoterin; however, amphoterin secretion was induced by androgen deprivation. The conditioned medium of human prostatic stromal cells deprived of androgen recovered the in vitro invasive capacity of PC 3 cells suppressed by amphoterin antisense S-ODN. These results suggest that androgen deprivation provides a paracrine interaction between cancer and stromal cells through the RAGE-amphoterin system in advanced prostate cancer. PMID- 14534710 TI - The inhibition of growth and angiogenesis in heterotransplanted esophageal carcinoma via intratumoral injection of arsenic trioxide. AB - To investigate the antitumor action of arsenic trioxide (As2O3) by intratumoral injection into solid tumors, tumor growth inhibition (TGI) and angiogenesis of heterotransplanted esophageal carcinoma in mice was carried out. The cultured human esophageal carcinoma cells were inoculated into both laterals of the abdominal wall of severe combined immunodeficient (SCID) mice. When both lateral tumors had grown to about 10x8x5 mm(3), the right tumors were treated with an intratumoral injection of As2O3 in dosage of 1, 5 and 10 microg per day, respectively, for 10 days sequentially. Left tumors were treated with PBS (phosphate buffer solution) as control. The weight of transplanted tumor masses were measured and counted for TGI. The tissue of tumor, liver, kidney, heart, lung and brain was examined histopathologically and tumor tissues were examined by light- or electron-microscope. Ki-67 and CD34 were assessed by immunohistochemistry and positive nuclei of Ki-67 and microvessel density (MVD) labeled by CD34 were measured. The results revealed that on the 20th day after the first injection, As2O3-treated tumors were suppressed markedly as compared with the contrarily situated tumor, accompanied by a marked apoptosis and necrosis in tumor cells. The tissue of liver, kidney, heart, lung and brain was unaffected by As2O3. MVD in tumor tissue was decreased in the right side tumor with the significant difference in the 5 micro g and 10 micro g group (p<0.01). TGI was 5.80 (p>0.05), 58.66 (p<0.01) and 73.97% (p<0.01) in the 1, 5 and 10 micro g groups respectively, but 2.21% (p>0.05) in the control group. Conclusively, a repeated administration of As2O3 (5 and 10 microg x 10) induced an increase of tumor growth inhibition and decrease of angiogenesis in the solid tumor in tumor progressive periods. These results suggest that intra-tumoral injection of As2O3 may be investigated as a modality to treat some solid tumors. PMID- 14534711 TI - Evaluation of CD44 variant 6 expression and clinicopathological factors in pulmonary metastases from colon carcinoma. AB - Although relatively little is known about the molecular mechanisms underlying tumor progression, recently CD44 glycoproteins and the c-Met receptor tyrosine kinase have been identified as potentially important components of the metastatic cascade. CD44 is a family of transmembrane receptors generated from a single gene by alternative splicing and differential glycosylation. Important biological processes involving CD44 glycoproteins include cell adhesion, lymphocyte homing, hematopoiesis, tumor progression and metastasis. The precise mechanism via which CD44 promotes tumorigenesis have not yet been elucidated. We evaluated the expression of adhesion molecule CD44 variant 6 in pulmonary metastases from colorectal carcinomas and its correlation with clinicopathological parameters. Twenty patients were randomly selected from the patients who had undergone a resection of pulmonary metastasis from colorectal cancer. Formalin-fixed, paraffin-embedded archival specimens of tumor tissues and adjacent normal mucosa from these patients were the subjects of the present study. Immunoreactivity for CD44 was quantified. Specimens were considered positive if almost 25% of the neoplastic cells were stained. CD44 v6 expression was related to the interval between colon resection and metastases diagnosis, the number of pulmonary metastases, and the survival after lung resection. No statistical correlation was found between CD44 v6 positivity and disease-free interval after colon resection, number of metastases or 2-year survival after lung resection. Probably CD44 v6 is necessary and sufficient to confer metastatic potential to carcinoma cells increasing the migration capacity and participating in invasion via changes in adhesion to the extracellular ligands, but is not necessary to modify the clinical history of the metastases. Therefore the evaluation of CD44 v6 expression in lung metastases does not influence the therapeutic scheme. PMID- 14534712 TI - Bone marrow micrometastasis detected by RT-PCR in esophageal squamous cell carcinoma. AB - The clinical implications of bone marrow micrometastases (BMM) detected by RT-PCR in esophageal squamous cell carcinoma (ESCC) have not been elucidated. We evaluated the relation between the presence of BMM, both before and after surgery, and clinicopathologic findings in patients with ESCC. Bone marrow samples from 48 patients with ESCC were obtained from the iliac crest before and after surgery. After total RNA was extracted from each bone marrow sample, carcinoembryonic antigen (CEA)-specific RT-PCR was performed. BMM was detected by RT-PCR in 10 of the 48 patients. Four patients each had positive signals only before or only after surgery and 2 patients had positive signals both before and after surgery. There were no significant differences in clinicopathologic factors, including neoadjuvant therapy, between patients with BMM and without BMM. To date, the rates of recurrent disease in patients with BMM and without BMM are 80% (8/10) and 50% (19/38), respectively, a difference which is not significant. The 4-year survival rates of patients with BMM and without BMM are 10.0% and 47.3%, respectively. Recurrence and survival rates were poorer in patients with RT-PCR positivity, although the differences were not significant. A larger study is required to clarify the clinical impact of BMM. PMID- 14534713 TI - Independent action of serine/threonine protein phosphatase in ovarian cancer plasma membrane and cytosol during gonadotropin-releasing hormone stimulation. AB - Reversible serine/threonine protein phosphorylation catalyzed by kinases and phosphatases 2A (PP2A) plays a crucial role in cellular growth and differentiation. We attempted to determine the subcellular location of PP2A in ovarian cancer cells and its regulation by gonadotropin-releasing hormone (GnRH), which is known to have anti-proliferative actions on ovarian cancers. Surgically removed ovarian cancers were screened for GnRH receptor expression prior to subcellular fractionations. PP2A activity was assessed by measuring the dephosphorylation of phosphopeptide highly selective for the PP2A in cytosol and membranes fractionated on a continuous sucrose density gradient. To assess GnRH effects, cloned cell lines were pretreated with or without a GnRH agonist. There were three peaks of PP2A activity, corresponding by marker enzyme analysis to the cytoplasm, plasma membrane and endoplasmic reticulum fractions. The kinetic analysis showed a different activity in cytosol and membrane; Km values for substrate of 185 microM and Vmax of 555 pmol/mg protein/30 min for cytosol, and 28 microM and 83 pmol/mg protein/30 min for plasma membrane, respectively. PP2A specific inhibitor okadaic acid inhibited the cytosolic and membrane-associated activity by 50% when added at 2 nM and 50 nM (p<0.001). A 50% inhibitory effect of NaF was obtained at 0.5-1 mM for cytosol and 5 mM for membranes (p<0.001). In Caov-3 cells exposed GnRH, PP2A activity of plasma membrane increased by 1.3-fold (p<0.001) but that of cytosol was not affected. PP2A activity in the plasma membrane of ovarian cancer cells might be distinct from that present in the cytosol. The plasma membrane PP2A may be responsible for a portion of an increased ser/thr protein phosphorylation-dephosphorylation turnover that occurs during cell exposure to GnRH. PMID- 14534714 TI - Protein and gene expression of tumor-associated antigen RCAS1 in esophageal squamous cell carcinoma. AB - RCAS1 (receptor-binding cancer antigen expressed on SiSo cells) is expressed on the tumor cell membrane and induces apoptosis on infiltrated immune lymphocytes. RCAS1 has been reported to correlate with the escape of tumor cells from host immune surveillance, and with poor prognosis. However, the clinical importance of RCAS1 protein and gene expression in esophageal squamous cell carcinoma (ESCC) has not been well investigated. In the present study, RCAS1 gene and protein expression levels were evaluated and compared with clinical findings in 67 patients with ESCC. Expression levels of RCAS1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) messenger RNAs (mRNAs) from tumors and non-cancerous epithelia were analyzed quantitatively by real-time reverse transcriptase polymerase chain reaction (RT-PCR). RCAS1 protein expression was analyzed by immunohistochemistry. The mean RCAS1/GAPDH ratio of tumors (0.7) was not different from that of non-cancerous epithelia (0.7, p=0.715). RCAS1 immunoreactivity was detected in 19 tumors (28.4%). The mean RCAS1/GAPDH ratio of tumors with RCAS1 protein positive (0.6) did not differ from tumors without RCAS1 expression (0.8, p=0.131). RCAS1 gene and protein expressions did not correlate with tumor size, depth of invasion, lymph node metastasis, or patient prognosis. Thus, RCAS1 gene or protein expression may not correlate with tumor progression in ESCC. PMID- 14534715 TI - beta-carotene supplementation in patients radically treated for stage I-II head and neck cancer: results of a randomized trial. AB - This study was aimed at evaluating the efficacy of beta-carotene in improving survival (S) and in disease-free survival (DFS) and reducing the incidence of second primary tumors (SPT) in patients with a radically treated stage I-II squamous head and neck tumors. Eligible patients were randomly allocated to receive beta-carotene (n=104) or no treatment (n=110). beta-carotene was administered at the dose of 75 mg/day for 3-month cycles within one month intercycle intervals for a 3-year period. The 3-year compliance to the beta carotene was 68.7%. Only eight patients reported drug-related toxicity (7.8%). The median follow-up of all patients was 59 months. The median follow-up was 61 months (range 1-116 months) in the beta-carotene and 58 months (1-123 months) in the control group. The 10-year DFS was 75.7% for the patients in the beta carotene and 74.3% for those in the control group (P=0.56). The 10-year S was 85.9% in the beta-carotene group and 80.9% in the control group (P=0.20). beta carotene supplementation had no significant effect on the incidence of second primary tumors (RR=0.99; 95% C.I. 0.28-3.44). A statistically non-significant 40% reduction in the risk of death among subjects assigned to the beta-carotene compared to the controls was observed (RR=0.60; 95% C.I. 0.26-1.38). No increase in the death from cardiovascular diseases was observed among patients treated with beta-carotene. Our results might support the hypothesis that an adequate beta-carotene treatment could be potentially associated with a decreased risk of death in these patients. PMID- 14534716 TI - Higher processing rates of Alu-containing sequences in kidney tumors and cell lines with overexpressed Alu-mRNAs. AB - Tumor cell growth and differentiation involve several molecular mechanisms that control gene expression and define specific genomic molecular profiles in cancer cells. Among these mechanisms, it has been shown that Alu-repetitive sequences are capable of regulating gene expression at transcriptional and posttranscriptional levels, and also of modulating cellular growth, differentiation and tumor suppression. Furthermore, repetitive sequences have also been implicated in alternative RNA splicing, although the specific mechanisms involved remain unknown. Nonetheless, exactly what the involvement of Alu-containing sequences in tumor cell growth and differentiation is or to what extent they might be related to tumorigenesis or to alternative splicing is not yet clear. In order to address some of these issues, we analyzed the level of expression of Alu-containing sequences in renal tumors and cell lines and their association with immunoprecipitated ribonucleoprotein splicing complexes in nuclear RNA fractions. Over-expression of Alu-containing sequences was detected in the poly(A)-RNA fractions of all analyzed tumors and cell lines. Furthermore, Alu-sequences were associated with tumor cell growth and differentiation and found overexpressed in purified small nuclear ribonucleoprotein fractions. Overall, our results suggest the involvement of Alu-sequences in the overexpression of Alu-containing-mRNAs in human tumors, and also higher processing rates of Alu-containing sequences at the spliceosome associated with tumor cell growth and differentiation. PMID- 14534717 TI - Tissue harmonic imaging sonography of the axillary lymph nodes: evaluation of response to neoadjuvant chemotherapy in breast cancer patients. AB - In breast cancer patients, the number of surgically resected metastatic axillary lymph nodes has been considered to correlate with prognosis. Therefore, the lymph nodes' response to chemotherapy may provide a favorable prognosis by suggesting an appropriate regimen of post-operative chemotherapy. In the present study, we evaluated the therapeutic efficacy of neoadjuvant chemotherapy of the axillary lymph nodes of breast cancer patients, using tissue harmonic imaging ultrasonography. Thirty-three female patients with breast cancer >2 cm in the longest diameter were examined by tissue harmonic imaging both before and following neoadjuvant chemotherapy. Nodes were defined as metastasis positive or negative on the basis of Yang's criteria, at each ultrasonography examination. The ultrasonography findings were correlated with the histopathologic results of the surgically resected specimens. Patients with constantly positive nodes over two ultrasonography examinations had a higher pathological nodes positive rate (8/9, 88.9%) than those (3/10, 30.0%) whose lymph nodes shrunk from the first to second ultrasonography exam (p=0.02, Fisher's exact method). All patients with negative nodes at both ultrasonography examinations had negative pathological results. Thus, changes in consecutive tissue harmonic imaging findings may be used to accurately evaluate the response of axillary lymph nodes to neoadjuvant chemotherapy. PMID- 14534718 TI - Rituximab in combination with platinum-containing chemotherapy in patients with relapsed or primary refractory diffuse large B-cell lymphoma. AB - The aim of the study was to evaluate the efficacy of a regimen consisting of rituximab and a platinum-containing chemotherapy with either Ifosfamide, Carboplatin and Etoposide (ICE) or Cisplatin, high-dose Ara-C and Dexamethasone (DHAP) in patients with relapsed or primary refractory diffuse large B-cell lymphoma. Ten patients with relapsed or primary refractory diffuse large B-cell lymphoma were treated from June 2000 until May 2001 with a platinum-containing chemotherapy regimen according to the ICE- or DHAP-protocol in combination with rituximab at the University of Muenster. Two cycles of ICE or DHAP and rituximab were given. In case of at least a minor response after 2 cycles, 2 additional cycles of the same combination were applied. Response rate, remission duration and duration of survival were evaluated. All 10 patients could be analysed with respect to these endpoints. No treatment related mortality was observed. The response rate (CR/PR) was 60% (10/50%). Twenty percent of the patients had progressive disease. The median duration of remission and survival was 3 and 3.5 months, respectively (range: 1-6 and 1-7 months, respectively), the survival rate was 10%. Eight of 10 patients died because of their underlying disease with short remission duration, 1 patient died of complications of allogeneic transplantation in CR. In conclusion, the combination of platinum-containing chemotherapy (ICE or DHAP) with rituximab demonstrates significant activity in intensively pretreated patients with relapsed or primary refractory diffuse large B-cell lymphoma. Considering the short duration of remission and survival, respectively, other experimental therapeutic approaches (e.g. allogeneic stem cell transplantation, radioimmunotherapy) should be pursued following this treatment in order to induce long-term remission. PMID- 14534719 TI - CD44 variant exon 6 expressions in colon cancer assessed by quantitative analysis using real time reverse transcriptase-polymerase chain reaction. AB - CD44 is a family of transmembrane glycoproteins that serve as a major receptor for hyaluronate and the splice variants play a very important role in tumor progression and metastasis. We examined the relationship between cancer progression and mRNA levels of CD44 variant exon 6 (CD44v6) in specimens of colon cancer at different diagnostic stages from 31 patients using real time RT-PCR analysis. Increased mRNA levels of CD44v6 were observed in 82% of the specimens in comparison with those in the corresponding non-cancerous tissue specimens. A statistically significant correlation between the CD44v6 expression and the cancerous state was found in most specimens at all Dukes stages. None of the other parameters were related to the expression in the cancerous specimens. Quantitative real time RT-PCR analysis showed that there was no correlation of CD44v6 expression with tumor progression, although CD44v6 is upregulated in transformation. Thus, CD44v6 expression may be a clinically useful indicator of colon cancer. PMID- 14534720 TI - Death receptor-dependent and -independent pathways in anticancer drug-induced apoptosis of breast cancer cells. AB - Apoptosis is a crucial event for anticancer drug efficacy. The signal pathways activated by anticancer drugs are classified as death receptor (DR) -dependent or -independent. The mechanisms by which anticancer drugs induce apoptosis via DR dependent pathways are not fully understood. In the present study, we assessed differential activation of signal transduction pathways leading to apoptosis by anticancer drugs in breast cancer cell lines. Three breast cancer cell lines, MDA MB-231, MCF-7, and MCF-7ADM, which is drug-resistant, were used. Drug sensitivity and apoptosis were assessed by MTT assay and TUNEL, respectively. Expression of apoptosis-related protein was assessed by Western blotting and RT-PCR. The sensitivities of MDA-MB-231 and MCF-7 cells to mitomycin C (MMC) and adriamycin (ADM) were similar. In contrast, sensitivity of MDA-MB-231 cells to paclitaxel (TXL) was 30-fold greater than that of MCF-7 cells, 0.03 micro M in MDA-MB-231 and 0.9 micro M in MCF-7 cells, respectively. Treatment with MMC increased expression of DR4 and Fas in a time-dependent manner in MDA-MB-231 cells. Treatment with ADM increased expression of DR4 and DR5 but not Fas, whereas treatment with TXL increased expression of Fas but not DR4 and DR5 in MDA-MB-231 cells. Although treatment of MCF-7 cells with ADM increased expression of DR4 and DR5 but not Fas, expression of DR4, DR5, and Fas by the drug-resistant cells did not change following treatment with ADM. Activation of Fas, DR4, and DR5 in drug sensitive cells in response to anticancer drugs is dependent on the cytotoxic effect of each drug. In drug-resistant cells, apoptosis is induced via DR independent pathways mediated by mitochondrial dysfunction. PMID- 14534721 TI - The cytotoxicity of hydrophobic bile acids is ameliorated by more hydrophilic bile acids in intestinal cell lines IEC-6 and Caco-2. AB - Bile acids, especially those with hydrophobic properties, are known to possess cytotoxicity. However, the mechanisms responsible for the cytotoxicity of bile acids are still under investigation. On the other hand, the hydrophilic bile acid, ursodeoxycholic acid has been reported to exhibit therapeutic effects against cytotoxic hydrophobic bile acids. The aim of the present study was to investigate the cytotoxicity of individual bile acids and combinations of bile acids using the intestinal cell lines IEC-6 and Caco-2 cells. The cytotoxicities of individual bile acids and the effects of various bile acid combinations were evaluated using the MTS [3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy phenyl)-2-(4-sulfophenyl)-2H-tetrazolium] assay. The bile acids induced cytotoxic effects depending on their hydrophobicity except for hyodeoxycholic acid. In the study for the effects of combined bile acids, not only ursodeoxycholic acid but other hydrophilic bile salts such as cholic acid and hyocholic acid exhibited cytoprotection against deoxycholic acid-induced cytotoxicity. Moreover, even some hydrophobic bile acids, such as chenodeoxycholic acid also exhibited cytoprotection. It is possible that the cytotoxicity of hydrophobic bile acids is ameliorated by more hydrophilic bile acids under certain conditions. The understanding of the precise mechanism of this phenomenon remains to be determined. PMID- 14534722 TI - Clinical significance in molecular detection of p53 mutation in serum of patients with colorectal carcinoma. AB - Circulating DNA can be isolated from serum of patients with various carcinomas and p53 mutation can be observed in colorectal carcinoma. The aim of this study was to investigate the correlation between p53 mutation in DNA extracted from colorectal carcinoma and that in DNA extracted from serum of patients with colorectal carcinoma. The clinical significance in molecular detection of p53 mutation in serum of patients with colorectal carcinomas was also investigated. DNA was extracted from tumors and non-tumorous colorectal tissues of 46 patients with single sporadic colorectal carcinomas of stage I (n=6), stage II (n=18), stage III (n=15), and stage IV (n=7) according to the TNM classification. Circulating DNA was also extracted from the serum of the 46 patients with colorectal carcinoma and from 7 healthy volunteers for normal control. Mutations of the p53 gene were analyzed using a fluorescence-based polymerase chain reaction (PCR)-single-strand conformation polymorphism (SSCP) method. DNA sequences were determined in DNA fragments with shifted peaks by SSCP methods. Mutations in tumors were found in 22 (48%) of 46 patients, and mutations in serum were found in 3 (14%) of these 22 patients. Of 4 patients with stage IV disease, 3 (75%) had serum p53 mutation and the mutation pattern of these 3 patients was the same in both tumor and serum. No correlation was seen between p53 mutation in serum and the level of serum DNA. There was no significant difference between the presence of p53 mutation in serum and tumor size, depth of invasion, vascular invasion, or lymph node metastasis. However, liver metastasis showed significant difference (p=0.0026). The presence of p53 mutation in serum was associated with a clinically advanced stage accompanied by liver metastasis. PMID- 14534723 TI - Detection of a novel alteration of the Axin gene in various pediatric neoplasms. AB - The Wnt signaling pathway is conserved in various species from worms to mammals, and plays critical roles in development, cellular proliferation and differentiation. As part of Wnt signal transduction, the function of the Axin complex is inhibited, leading to the accumulation of beta-catenin protein. Axin gene mutations have been detected in several kinds of human cancers. In this study, we investigated Axin gene alterations in a series of 58 pediatric neoplasms including neuroblastomas, teratomas, rhabdomyosarcomas. Forty-eight non tumor tissues were used as a control series to compare gene alterations and their frequencies between tumors and normal tissues. The whole coding region of the Axin gene was examined by PCR-SSCP method using 24 sets of the primers. Samples revealing aberrant band patterns were subjected to direct sequencing analysis. In total, we identified six variants in the exons and four intronic nucleotide substitutions in the tumor series. Similar sequence variants except a rare sequence variant at codon 98 (CCG right curved arrow CTG) were observed in the control series and these were regarded as non-pathogenetic polymorphisms. Our results indicated that a tumor-associated mutation of the Axin gene is generally a rare event in our series of pediatric neoplasms. PMID- 14534724 TI - Effects of tumor necrosis factor alpha in renal cell carcinoma. AB - Tumor necrosis factor alpha (TNFalpha) and TNFalpha receptor mRNA expression, the effects of TNFalpha on DNA synthesis and cell proliferation as well as its effects on interleukin-6 (IL-6) production and expression in renal cell carcinoma (RCC) were studied using RCC cell lines. The effects of TNFalpha on DNA synthesis and IL-6 production were also studied using short-term established RCC cell lines. A total of 8 cell lines, 4 RCC cell lines (RC-2, RGB, 14T, 4T) and 4 cell lines established at our laboratory (OCUU-1, 2, 4, 5), as well as 10 short-term established RCC cell lines were used. When TNFalpha and TNFalpha receptor mRNA expression was examined by RT-PCR, p55 TNF receptor mRNA expression was observed while TNFalpha and p75 TNF receptor mRNA expression was not observed in all cell lines. When the effects of TNFalpha on DNA synthesis were studied by [3H] thymidine incorporation assay, DNA synthesis was induced in RGB, 14T, 4T, OCUU-2 and OCUU-4 by adding 10 and 100 pg/ml of TNFalpha while it was not induced in RC 2 and OCUU-5 at all concentrations. When its effects on cell proliferation were evaluated by MTT assay, cell proliferation was observed in RGB, 14T and 4T at TNFalpha concentration of 10 pg/ml and in RGB, 14T, 4T, OCUU-4 and OCUU-5 at TNFalpha concentration of 100 pg/ml. However, cell proliferation was not detected in RC-2 and OCCU-2 at any concentration. When the effects of TNFalpha on IL-6 production were studied by ELISA, IL-6 production was induced in RC-2, RGB, 14T, OCUU-1, OCUU-2 and OCUU-5 while not in 4T and OCUU-4. When its effects on IL-6 expression were examined by Northern blot analysis, the results were similar to those obtained by ELISA. As for the 10 short-term established RCC cell lines, DNA synthesis and IL-6 production were induced with the addition of TNFalpha. These results suggested that TNFalpha induced cell proliferation in RCC. PMID- 14534725 TI - Case report: recurrence of soft tissue MFH in bone due to minute intravenous tumor emboli detected by MRI. AB - We recently encountered a case with local recurrence of malignant fibrous histiocytoma (MFH) in the bone after wide resection, caused by minute intravenous tumor emboli which were retrospectively detected in MR imaging. The patient was a 69-year-old woman who initially noticed a mass in her left thigh. The tumor was diagnosed to be MFH, therefore a wide resection was performed; although the tumor was closely attached to the periosteum, it was not difficult to dissect the tumor subperiosteally from the cortex of the femur. The patient received postoperative brachytherapy, but no chemotherapy. Two years later, the tumor recurred with bony destruction of the femur. We reviewed the pre-operative films obtained by various imaging modalities, as well as the histology of the primary tumor, and found minute intravenous tumor emboli in the MR imaging obtained before surgery. Tumor emboli were also observed histologically in the small vessels of the surgically resected tumor. Such intravenous tumor emboli have recently been implicated in the development of regional bone metastasis near the site of the primary lesion in cases of malignant soft tissue tumors. Therefore, we concluded that the tumor recurrence in our case was caused by small tumor emboli invading the perforating veins of the femur. It is therefore emphasized that MR images should be carefully reviewed for the presence of such intravenous tumor emboli before surgery in cases of high-grade malignant sarcomas. As at the time of writing, our patient remains alive and disease-free, with no evidence of any local recurrence or distant metastasis after wide tumor resection for the recurrent tumor. PMID- 14534726 TI - Effect of retinoblastoma tumor suppressor gene expression on chemosensitivity of human osteosarcoma cell lines. AB - We examined the effects of inactivation of the RB gene on chemosensitivity of human osteosarcoma cell lines, using the MTT assay and calculating the inhibition index. Although the human osteosarcoma cell lines HOS and MG63 have a wild-type RB gene, SaOS-2 and OSrb (established from retinoblastoma patient) have no active RB gene. We used these 4 cell lines in growth inhibition assays for doxorubicin, cisplatin and methotrexate, and assessed the chemosensitivity. In the growth inhibition assay for methotrexate, cell lines lacking an active RB gene were more resistant than cell lines with an active RB gene. PMID- 14534727 TI - The effect of epidermal growth factor receptor antisense morpholino oligomer on non-small cell lung cancer cell line. AB - Overexpression of the epidermal growth factor receptor (EGFR) has been identified as a common component of various cancer types including lung cancer. Recently morpholino oligonucleotides appeared as a promising modification for antisense applications with few toxic effects and their stability. We investigated the effect of EGFR antisense morpholino oligomer on non-small cell lung cancer (NSCLC) cell line by evaluating EGFR mRNA, protein product and cell proliferation. The EGFR antisense morpholino oligomer was designed to target the translation start site in the EGFR mRNA. The four base-mismatch morphlino oligomer was designed as a control for EGFR antisense morpholino oligomer. These morpholino oligomers were introduced into NCI-H125 cell line which showed overexpression of EGFR. The EGFR mRNA and protein expression were quantified by real time RT-PCR and ELISA, respectively. The significant repression in both EGFR mRNA and protein expression was observed for three days after single treatment with EGFR antisense morpholino oligomer. Furthermore, the growth of NCI-H125 cell line was significantly inhibited with treatment by EGFR antisense morpholino oligomer. Our results indicate that EGFR antisense morpholino oligomer represses the EGFR expression at both mRNA and protein level and inhibits the proliferation of NSCLC cell line suggesting that it may be a promising strategy as one of antisense therapies for NSCLC. PMID- 14534728 TI - Pituitary stalk metastasis from breast cancer treated with systemic chemotherapy. AB - We describe a 61-year-old woman with diabetes insipidus caused by a pituitary stalk metastasis from breast cancer. She had a medical history of breast conservation therapy for early breast cancer 5 years previously. Pituitary, lung, liver, bone and neck lymphnode metastases was revealed at the same time. She received systemic chemotherapy consisting of docetaxel and cisplatin. After chemotherapy MRI finding in pituitary gland was improved, lung and liver metastases also improved, however, symptoms of diabetes insipidus did not improve. She is alive, receiving endocrine treatment for 2 years, since onset of diabetes insipidus. PMID- 14534729 TI - The relative contributions of carboxylesterase and beta-glucuronidase in the formation of SN-38 in human colorectal tumours. AB - Irinotecan (CPT-11) is a prodrug that is used to treat metastatic colorectal cancer. It is activated to the topoisomerase poison SN-38 by carboxylesterases. SN-38 is subsequently metabolised to its inactive glucuronide, SN-38G, which can however be reactivated to SN-38 by beta-glucuronidase. The purpose of this study was to examine the role of carboxylesterases and beta-glucuronidase in the in vitro production of SN-38 in human colorectal tumours. The production of SN-38 from CPT-11 and SN-38G was measured by HPLC in human colorectal tumour homogenates. Carboxylesterase and beta-glucuronidase activities were found to be lower in tumour tissues compared to matched normal colon mucosa samples. In colorectal tumour, beta-glucuronidase and carboxylesterase-mediated SN-38 production rates were comparable at clinically relevant concentrations of SN-38G and CPT-11, respectively. Therefore, tumour beta-glucuronidase may play a significant role in the exposure of tumours to SN-38 in vivo, particularly during prolonged infusions of CPT-11. PMID- 14534730 TI - Evaluation of MUC1 splice variants as prognostic markers in patients with ductal carcinoma in situ of the breast. AB - Despite intense research in the field of breast cancer it still remains the most common cancer in women in the Western world. A decreasing trend in mortality was mainly achieved by improved early detection which led to an increased incidence of ductal carcinoma in situ (DCIS) of the breast. For the patient's prognosis and the administration of a patient-tailored therapy strategy it is crucial to identify diagnostic and prognostic markers for high-risk DCIS patients. MUC1 is associated with tumour aggressiveness in human breast cancer. Recent studies used MUC1 splice variant A to identify malignant thyroid cancer. In the present study we have examined the usefulness of MUC1 splice variants as prognostic markers in DCIS. We used laser capture microdissection of paraffin-embedded tissue to isolate RNA from isolated tumour cells and determined the MUC1 splice variant distribution by RT-PCR. In the majority of cases variant B was more highly expressed than variant A. This was true for pure DCIS (66%) as well as for DCIS with adjacent invasive cancer (66%). In 7 out of 18 cases (38%) of pure DCIS variant A was not expressed at all. In DCIS with adjacent invasive cancer only 2 samples out of 12 showed this expression pattern (16%). The situation that variant A was more highly expressed than B, or that variant B was not expressed at all, was similar for pure DCIS (27%) and for DCIS with adjacent invasive cancer (33%). The present study describes the differences of MUC1 splice variant expression in pure DCIS compared to DCIS with adjacent invasive cancer. A discriminating pattern of MUC1 splice variants could not be demonstrated. PMID- 14534732 TI - Long-term cryopreservation of peripheral blood stem cells in patients with advanced germ cell tumors using the dump-freezing method at -80 degrees C. AB - The objectives of this study were to evaluate the hematopoietic activity of peripheral blood stem cells (PBSCs) harvested from patients with advanced germ cell tumors after long-term cryopreservation in a mechanical freezer at -80 degrees C. Forty-one patients with advanced tumors were enrolled in this study. Flow cytometric analysis was performed to determine the total cell viability and the viability of CD34 positive cells of PBSCs stored between January 1995 and December 2001. The 41 patients were divided into two groups according to the duration of cryopreservation. In the 21 patients in whom the duration of storage was greater than 5 years, the total cell number did not change significantly before and after long-term cryopreservation (p=0.75). The percentage of viable cells and the total number of viable cells was significantly decreased after long term cryopreservation (p<0.001 and p<0.05, respectively). However, the percentage of CD34 positive cells was significantly increased to 2.34% from 1.85% (p<0.05), resulting in non-statistically significant change in the total viable CD34 positive cells (p=0.98). A similar relation was found in the 20 patients whose storage duration was less than 2 years, although a significant change was found only in the percentage of viable cells. Moreover, pre-PBSCH factors, including age, cumulative dosage of cisplatin and the International Germ Cell Cancer Collaborative Group classification at the diagnosis did not affect the viable CD34 positive cell count after long-term cryopreservation. In conclusion, the present results suggest that CD34 positive cells could be successfully cryopreserved in a mechanical freezer at -80 degrees C for more than 5 years with no relation to pre-PBSCH factors and efficacy of PBSCH itself. PMID- 14534731 TI - Genetic and biochemical changes in colorectal carcinoma in relation to morphologic characteristics. AB - A distinct genetic pathway may be involved in the development of polypoid and flat colorectal cancers, two morphologically different cancer subtypes. The present study was undertaken to clarify whether different combinations of some genetic alterations commonly involved (such as K-ras and p53 gene mutations) may exist between polypoid and flat types. In addition, to investigate any different proliferative behavior between the two distinct types of colorectal cancer, we tested the enzymatic activity of ornithine decarboxylase (ODC). A total of 29 polypoid type and 21 flat type colorectal cancers were selected for this study. We investigated K-ras and p53 mutations by polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP) and single strand conformational polymorphism (PCR-SSCP) analysis, respectively. A radiometric method was used to evaluate ODC activity. K-ras and p53 gene mutations were present in 30 and 48% of cases, respectively. A significant association between the p53 mutation and the flat type of colorectal cancer was detected; on the contrary, no significant difference in frequency of K-ras mutation between polypoid and flat type colorectal cancer was found. A statistically significant difference in ODC activity levels was observed between polypoid and flat types. Moreover, we found that ODC activity was significantly higher in neoplastic tissue than in surrounding normal mucosa in polypoid type colorectal cancer. Different mutation patterns and proliferative behavior were observed in polypoid and flat colorectal malignant tumors. Further studies will be required to ascertain whether the distinct growth appearance of colorectal cancer can affect the outcome and prognosis of patients with this type of malignant disease. PMID- 14534733 TI - Angiogenesis is continuous with two peaks during azaserine-induced rat pancreatic adenocarcinoma progression: an electron microscopic morphometrical study. AB - Although the indispensable role of neoangiogenesis in tumour growth became generally accepted, quantitative microvascular changes associated to tumour progression have been followed only in a few experimental models. In our study a further experimental system, the azaserine-induced rat pancreatic adenocarcinoma was applied for this purpose. The 15-20 month long progression of this tumour provides us a special opportunity to dissect multistage carcinogenesis in different respects. Tumour samples were taken at months 6, 8 (early lesions), 10, 13, 15 (adenomas), and 20 (differentiated and anaplastic adenocarcinomas) and identified by semithin section histology. We applied reliable and reproducible electron microscopic morphometry for quantification of capillary volume and surface densities in unit tissue volume. Correlation of microvessel volume (MVVD) and surface (MVSD) density data indicated that their changes were the consequence of the changing balance between tumour growth and neoangiogenesis. The significantly decreased but persistent size of the tumour microvasculature during the early slow premalignant growth (months 6-10) is indicative for a slow, but continuous balanced angiogenesis. A dramatic MVVD and MVSD increase at month 13 and the consecutive decrease measured in month 15 adenomas depicted a hitherto unprecedented angiogenic wave within the month 10-15 premalignant growth period. The size of the tumour microvasculature became again stabilized below the control level for the next 5 months of slow growth during which, however, several differentiated adenocarcinomas were transformed to fastly growing angiogenic anaplastic carcinomas, some of which gave rise to invasive tumours. These findings once again verify the angiogenic switch theory. PMID- 14534734 TI - Tumour MHC class I downregulation and immunotherapy (Review). AB - MHC class I downregulation is an important mechanism of tumour escape from T cell mediated immune responses. Approximately 40-90% of human tumours derived from various MHC class I+ tissues were reported to be MHC class I deficient. Decreased or absent MHC class I expression is frequently associated with the invasive and metastatic tumour phenotype. Altered MHC class I antigen expression involves total loss, loss of haplotype, locus downregulation, allelic loss or downregulation, and combinations. Description of partial or complete losses of MHC class I molecules in tumour cells as mechanisms of immune escape often fails to consider an increased susceptibility to NK cell-mediated lysis, which is a direct consequence of such losses. A low MHC class I level favours NK cells as effectors, whereas a high level of MHC class I favours T cells as effectors. The microheterogeneity of MHC class I expression in tumour cell populations, the balance of the MHC-restricted and MHC-unrestricted defence as well as the selective pressure of antigen-specific (CTL) and antigen non-specific (NK) effector mechanisms decide the final outcome of the MHC class I expression in the primary tumour and its metastases as well as the final outcome of the tumour defence reaction. Despite the MHC class I molecule deficiency and the resulting absence of the CD8+ T cell-mediated immunity, the tumour hosts were found to be capable of being immunized against MHC class I- tumours. The purpose of this review is to discuss the positive results of MHC class I- tumour treatment obtained with immunomodulatory cytokines and tumour vaccines, as well as the prospects and limitations of such therapy. PMID- 14534735 TI - In vitro evaluation of chemopreventive agents using cultured human prostate epithelial cells. AB - The effects of nine potential cancer chemopreventive agents on cell growth or clonogenic survival were evaluated in normal human prostate epithelial cells, an immortalized but non-tumorigenic human prostate epithelial cell line (267B1), a human benign prostatic hyperplasia (BPH) cell line (BRF-55T), and a human prostate cancer cell line (267B1/Ki-ras). Of the nine agents tested, 9-cis retinoic acid, liarozole fumarate, phenylenebis(methylene)-selenocyanate (p-XSC), and L-selenomethionine demonstrated much stronger growth inhibitory effects on prostate cancer cells than on the normal prostate epithelial cells, suggesting that these agents may be useful as prostate cancer chemopreventive agents. 9-cis retinoic acid, genistein, liarozole fumarate, p-XSC, L-selenomethionine and vitamin E also showed much stronger growth inhibitory effects on BRF-55T cells than on the normal prostate epithelial cells, indicating that these agents may also be useful for the prevention and treatment of BPH. Difluoromethylornithine (DFMO), DHEA analogue 8354 (fluasterone), and oltipraz did not show strong inhibitory effects on the growth or survival of normal prostate epithelial cells, 267B1 or 267B1/Ki-ras cells, suggesting that these agents may not be effective as prostate cancer preventive or therapeutic agents. PMID- 14534736 TI - Plasma leptin and colorectal cancer risk: a prospective study in Northern Sweden. AB - Obesity is associated with an increased risk of colorectal cancer. Circulating levels of leptin are high in obesity and strongly correlated to levels of insulin. Leptin stimulates growth of colon cancer cells. In a nested case-control study, we measured leptin levels in prediagnostic plasma from 75 men and 93 women who were diagnosed with colorectal cancer mean time 3.4 years (SD 2.4) after recruitment and among 327 control subjects. Logistic regression analyses showed increases in colorectal cancer risk in men with increasing levels of leptin, odds ratios (OR) were 1.00 (ref), 0.85 (95% C.I.=0.33-2.23), 1.04 (0.43-2.53), and 2.15 (0.89-5.22), (pfor trend=0.08). There was a distinct threshold between the third and fourth quartile of leptin, and the odds ratio for top quartile vs. three bottom quartiles was 2.28 (1.09-4.76). Adjustment for body mass index and insulin did not affect risk estimates. In separate analysis, odds ratio for top vs. bottom tertile of colon cancer was 1.96 (95% C.I.=0.72-5.29), whereas no increase was seen for rectal cancer. In women, no association between leptin and risk was seen. These data support the hypothesis that leptin is a risk marker for colorectal cancer in men, but not in women. PMID- 14534737 TI - Potentiation of the antiproliferative activity of MKT-077 by loperamide, diltiazem and tamoxifen. AB - MKT-077, a delocalized lipophilic cation, selectively targets cancer cells. MKT 077 has been reported to inhibit the growth of several tumor types and has undergone phase I clinical testing. We have examined the effect of MKT-077, alone and in combination with the antidiarrheal drug loperamide. Ten human cancer cell lines, four prostate (PC3, DU145, LNCaP, MDA-PCA-2B), two breast (MCF-7 and MDA MB-231) and four colon (LoVo, Colo320DM, SW1116 and LS174t) were tested in vitro. Cells were grown to confluency prior to treatment. Loperamide potentiated the antiproliferative effect of MKT-077 in all ten cell lines, in a dose-dependent manner. The sensitivity of MDA-PCA-2B cells, the two breast and four colon cancer cell lines to MKT-077 was relatively low (>2.5 microg/ml MKT-077 required to inhibit growth by 95%). In these cell lines, 0.5-5 microg/ml (1-10 microM) loperamide caused a marked increase in the response to MKT-077. Loperamide is known to activate micro-opioid receptors at nanomolar concentrations and block voltage-gated calcium channels at micromolar doses. We found that calcium channel blockers diltiazem and nifedipine (10-20 microg/ml), as well as tamoxifen (1.5 2.5 microg/ml) can also potentiate the growth-inhibitory effects of MKT-077. These synergistic interactions could be exploited for therapeutic benefit. PMID- 14534738 TI - Aphidicolin decreases ex vivo resistance to cytosine arabinoside in childhood acute leukaemia. AB - In order to assess the effect of the DNA polymerase inhibitor aphidicolin on resistance to cytosine arabinoside, blast cells from 15 children with ALL and 9 with AML were exposed to a range of concentrations of ara-C +/- aphidicolin. Cell survival was measured using the MTT assay. Aphidicolin significantly increased sensitivity to ara-C in blast cells from both ALL (p=0.001) and AML (p<0.01). The median fold increase (sensitisation ratio) for ALL was 3.4 (range 1.2-13.6) compared to 12.4-fold (range 6.0-148) for AML blasts (p=0.005). There was a striking relationship between increasing ara-C resistance and increasing effect of aphidicolin in AML (p<0.001) but not ALL (p>0.05). These remarkable results suggest that aphidicolin should be considered for future clinical trials as a modulator of ara-C resistance, particularly in AML. PMID- 14534739 TI - P53 mutations as an identification marker for the clonal origin of bladder tumors and its recurrences. AB - The clonality of synchronous and metachronous bladder tumors has been studied for years with controversial results. Some recent studies support the 'polyclonal origin' hypothesis, i.e. that independently transformed different tumor cell clones exist in the same bladder cancer patient and arise from the field cancerogenisation affecting the entire bladder urothelium by environmental mutagens. Others could demonstrate a monoclonal origin of primary bladder tumors and its recurrences due to a single genetically transformed cell clone spread through the urinary system. With increasing understanding of the clonal origin of bladder tumors and recurrences, clonality markers might contribute to an early and accurate prediction of tumor recurrence and progression. We used p53 mutations as an identification marker permitting the prediction of clonality in bladder tumors and its recurrences. Primary tumors (n=33) and recurrences (n=63) were screened by direct genomic sequencing the p53 mutation hot spot region, exons 5-8. P53 mutations occurred in 12% in our cohort, predominantly in higher malignant (>or=G2), invasive (>or=T1) tumor samples. We were able to demonstrate intratumoral heterogeneity regarding the p53 status and that recurrences may occur from genetically unrelated primary tumor sites. Some of our results argue for a polyclonal origin of synchronous and metachronous bladder tumors possibly due to the field effect in bladder carcinogenesis. Evidence for a monoclonal origin was found in two cases: one case with a high malignant primary tumor and 3 metachronous recurrences, all of them harbouring the same exon 8 mutation found in the primary tumor; one case with identical mutations of exon 8 in the primary and one recurrent tumor. For further implications concerning clonality of recurrent bladder tumors, p53 status should be combined with a broader range of markers such as CGH and LOH pattern. PMID- 14534740 TI - Expression of transglutaminases in human breast cancer and their possible clinical significance. AB - Implicated in several physiologic processes, including tumour-host, some transglutaminases (TGase) have been noted to play a regulatory role in the extracellular matrix in cell adhesion and migration of cancer cells. This study sought to determine the level of expression of TGases and their possible clinical significance in a cohort of human breast cancer patients using RT-PCR and quantitative RT-PCR. Normal breast tissues generally expressed low levels of TGases-1, 2, 3 and 7, and higher levels of TGases-4, 5 and plasma TGase (FXIII). Significantly increased levels of transcripts of TGases-4 and 7, and significantly lower levels of FXIII were seen in tumour tissues (n=110) compared with normal mammary tissues (n=27), p=0.05, 0.04 and 0.05, respectively. Node positive tumours exhibited significantly higher levels of TGase-2 and lower levels of TGase-3 (p=0.05 and 0.046, respectively). The lowest levels of TGases-3 and 7 were seen in patients with metastatic disease, and TGase-3 in patients who died of breast cancer, compared with those who remained disease-free (median follow-up 72 months). Higher levels of TGases-4 and 5 were noted in patients with local recurrence. Breast cancer displays an aberrant expression of TGases, wherein the levels of TGases-2, 3 and 7 have a relationship with node involvement and patient outcome. PMID- 14534741 TI - Sulforaphane and 2-oxohexyl isothiocyanate induce cell growth arrest and apoptosis in L-1210 leukemia and ME-18 melanoma cells. AB - Flow cytometry and laser-scanning confocal microscopy were used to study the effect of sulforaphane (SFN) and 2-oxohexyl isothiocyanate on the growth and viability of mouse leukemia L-1210 and human melanoma ME-18 cells during their exponential growth. Sulforaphane belongs to a group of compounds known as isothiocyanates. Isothiocyanates mainly occur in Cruciferous family. In particular, they occur in many vegetables such as broccoli and their sprouts. SFN and 2-oxohexyl isothiocyanate are potent inducers of detoxication phase 2 enzymes in mouse tissues and murine hepatoma cells in culture. Sulforaphane was shown to induce cell growth arrest in a dose dependent manner, followed by cell death. Sulforaphane induced the cell death via an apoptotic process. Two markers of apoptosis were investigated: phosphatidylserine externalization, which occurs in the early stages of apoptosis, and DNA strand breaks. Our results strongly suggest of chemopreventive activity toward cancer by the induction of apoptosis by SFN and 2-oxohexyl isothiocyanate. PMID- 14534742 TI - Older age and second skin cancer as prognostic factors in localized malignant melanomas. AB - Of the numerous prognostic factors for patients with localized malignant melanoma (LMM), none is superior to the simple parameter of tumor thickness. The aim of the present study was to better define prognostic factors for this disease. Between January 1992 and December 1994, 188 consecutive patients with LMM were treated at the Rabin Medical Center. Patient and tumor characteristics were retrospectively examined as potential prognostic factors. Patients (n=173) who had had at least two-year follow-up were included in the overall survival (OS) analysis, and 159 patients for whom accurate data on recurrent disease were available were included in the disease-free survival (DFS) analysis. At a median follow-up of 85 months (range 24-114), 48 patients (30%) had recurrent disease which resulted in death in 35 (20%). The five-year OS and DFS rates for the entire group were 82 and 72%, respectively. On univariate analysis, female gender, age younger than 75 years, metachronous or synchronous second skin cancer (including melanoma), light skin color, tumor thickness and TNM stage were predictive of both OS and DFS. Tumor location and ulceration, correlated with only one endpoint, OS or DFS, respectively. On multivariate analysis, three factors retained statistical significance with regard to both OS and DFS: tumor thickness (p=0.000 for both), second skin cancer (p=0.02 for both), and age (p=0.04 for both). Alongside the well-established predictive factor of tumor thickness in LMM, older age and the presence of a second skin cancer also have prognostic significance. The prognostic importance of the latter is reported here for the first time. PMID- 14534743 TI - Autologous soluble tumor-associated antigens prevent the toxic side effects of cancer chemotherapy and inhibit the progress of tumorigenesis: case report. AB - In this communication, we report for the first time, that immunization of cancer patients with autologous soluble tumor-associated antigens (sTAA) isolated from their own serum prevents the toxic side effects of chemotherapy, improves the patients' clinical status, and has therapeutic effects without chemotherapy. In 2001 and 2002, two cancer patients were treated, during chemotherapy, with autologous sTAA. Another benign tumor-bearing patient was treated with a medicinal herb and autologous sTAA. Doses for subcutaneous injections varied between 2.5 and 3 mg of sTAA in 0.5 ml of sterile distilled water. Injections were performed twice a week or at weekly intervals. In each case, the clinical status of the patient became more stable and healthier. Toxic side effects caused by chemotherapy decreased or even disappeared. No additional toxic side effects were observed after vaccination with sTAA. In the studied cases, a polyp disappeared and a metastatic brain tumor began to encapsulate. No metastases were seen in the case with colon adenocarcinoma. We concluded that vaccination of patients with autologous sTAA prevents the toxic side effects of chemotherapy in cancer patients and improves their clinical status. In the case with the benign tumor, this vaccination activated the host's immune system, prevented progress of the disease and even promoted tumor disappearance. We suggest that immunotherapy with autologous sTAA provides significant clinical benefits in cancer patients and appears to be an important new adjuvant treatment of cancer. PMID- 14534744 TI - Development of artificial chimerical gene regulatory elements specific for cancer gene therapy. AB - To achieve satisfactory outcome by the expression of therapeutic genes, it is of great importance to obtain efficient and high level of gene expression as well as minimizing inappropriate gene expression in non-target cells. To accomplish this goal for cancer gene therapy, we have evaluated the potential of cancer specific gene expression of functional promoter/enhancer elements in six putative cancer specific genes (Tcf1alpha, C-Ha-Ras, CyclinE, Cdc25A, HK II, and hTert) using a luciferase reporter assay. Most of the reporter constructs showed higher activity in HepG2 cells than in non-transformed or stem cells, and, in particular, the hTert (E) or Tcf1alpha (T) regulatory element showed significantly higher activity. We have also constructed a series of artificial chimerical regulatory elements by combinatorial linking of E promoter and T enhancer. A dramatic decrease of activity was observed as the copy number of concatenated T/E regulatory elements increased. In contrast, in chimerical constructs containing two or three copies of regulatory elements of T/E, cell type preferential expression profiles were changed. Thus, both pGL3-TE and -TEE showed higher activity specifically in MCF7 breast cancer cells, whereas pGL3-TET showed moderate activity in several cancer cell lines of different origins. Our results demonstrate that although the transcriptional activities of synthetic promoters are weak, some cancer-specific regulatory elements are useful in developing optimized and systemic cancer-specific regulatory regions with potential application in targeted cancer cell therapy. PMID- 14534745 TI - Artifactual formation of 8-oxo-2'-deoxyguanosine: role of fluorescent light and inhibitors. AB - The pro-mutagenic oxidative DNA lesion, 8-oxo-2'-deoxyguanosine (8-oxodG) has been a subject of numerous studies. However, the absolute 8-oxodG levels in tissue DNA reported by various methods have been debated due to its artifactual production during DNA isolation and/or the DNA processing. We have investigated factors that may result in such artifacts during isolation and analysis of DNA as well as means for its prevention. 8-OxodG content was measured by a recently described TLC enrichment-mediated 32P-postlabeling. Liver DNA from 3 month-old, female Sprague-Dawley rats was isolated by a standard solvent-extraction procedure (phenol, phenol:Sevag, and Sevag; 23 degrees C), a modified solvent extraction procedure (phenol:Sevag, and Sevag; 4 degrees C; KCl-SDS-protein precipitation) or sodium iodide extraction procedure. 8-OxodG was analyzed in the DNA by the 32P-postlabeling assay using a fluorescent light box during the workup, as well as in its absence. The 8-oxodG levels, when the fluorescent light box was used, were in similar range irrespective of the DNA isolation procedure (16.4+/-1.6 to 28.7+/-6 8-oxodG/10(6) nucleotides). However, the values were significantly lower (3.1+/-0.4 to 3.4+/-0.2 8-oxodG/10(6) nucleotides) in the absence of fluorescence light box, room fluorescent light (suspended through the ceiling) and natural room light did not alter the 8-oxodG levels. Further, the addition of 0.3 mM of PBN (N-t-butyl-alpha-phenyl nitrone) or TEMPO (2,2,6,6 tetramethylpiperidine-N-oxyl), or 6.8 mM 8-hydroxy-quinoline during the DNA isolation resulted in still lower values (0.8+/-0.1 to 1.8+/-0.5 8-oxodG/10(6) nucleotides) although this reduction was not consistently observed in different experiments. These results suggest that fluorescent light is the major 'culprit' in artifactual production and variability reported in the 8-oxodG levels. PMID- 14534746 TI - Immunohistochemical profile and treatment of uncommon types of thyroid carcinomas. AB - The distinction of uncommon types of thyroid carcinomas is important, because their treatment and prognosis differ. The aim of this study was to describe retrospectively the immunohistochemical profile of uncommon types of thyroid carcinomas and mode of treatment. Of the 1194 patients with thyroid carcinomas treated in Rabin Medical Center from 1954 to 2001, 153 were uncommon types (not papillary or follicular carcinomas). Specimens from archival tissue obtained from thyroidectomies in all these cases were revised and immunohistochemically examined. Anaplastic carcinomas (n=59) were positive for high molecular weight cytokeratin (CK HMW), low molecular molecular weight cytokeratin (CK LMW), cytokeratin (CK) 7, CK 8 and 18, thymoglobulin, EMA and vimentin; medullary carcinomas (n=39) were positive for CK LMW, CK 19, CK 8 and 18, CK 10, CK 7, carcinoembryonic antigen (CEA) and calcitonin; Hurthle cell carcinomas (n=30) for CK LMW, CK 19, CK 8 and 18, thyroglobulin, epithelial membrane antigen (EMA) and CEA; squamous cell carcinomas (SCC) (n=12) for CK HMW and cytokeratin total (CKs); lymphomas (n=7) for leukocyte common antigen (LCA) and B-cells (CD 20), and clear cell carcinomas (n=6) for CK LMW, CEA and thyroglobulin. Use of an immunohistochemical panel has diagnostic value in the differentiation of uncommon types of thyroid carcinoma, which help to plan the best mode of treatment. PMID- 14534747 TI - Radiotherapy for partially resected spinal ependymomas: a retrospective study of 60 cases. AB - Ependymomas (Es) and myxopapillary ependymomas (mpEs) are the most common primary tumours of the spinal cord. Recurrence-free survival depends on local control of the tumour. The value of additional radiotherapy (RT) is still a matter of debate. The aim of this retrospective study was to analyse radiotherapy, surgery and the preoperative state with regard to recurrence rate and long-term neurological outcome. Sixty patients with spinal Es (40) and spinal mpEs (20) were included. According to local policy, 14 patients who underwent total resection and 20 patients with incomplete resection were irradiated postoperatively. Total resection was achieved in 34 of the 60 tumours. Preoperative state and long-term outcome was assessed according to a previously published scale. When postoperative RT was applied after partial resection, only 3 of 11 Es and 1 of 9 mpEs recurred. All partially resected non-radiated Es (n=3) and 2 of the 3 partially resected non-radiated mpEs recurred. There was no recurrence after total resection. Only one of 6 patients with disseminated mpEs had clinical symptoms caused by the disseminated tumour. Long-term neurological outcome was related to preoperative conditions with no difference between partially and totally resected tumours. Our study shows that RT is only beneficial for partially resected Es and mpEs. Local recurrence-free survival of spinal Es and mpEs is obtained by total resection. Long-term neurological outcome is related to preoperative conditions. Seeding is seen in mpEs and does not cause clinical symptoms in most of the patients. PMID- 14534748 TI - Alterations in the activity and expression of serine/threonine protein phosphatases during all trans retinoic acid-induced apoptosis in breast cancer cells. AB - Retinoids exert different effects on malignant cells with various phenomena. They can induce differentiation and apoptosis in various cancer cells. However, the underlying mechanism of these effects is not clear. There are data related to the role of protein phosphatases during retinoid-induced leukemic cell differentiation. The aim of this study was to evaluate effects of the All trans retinoic acid (ATRA) on protein/serine phosphatases during ATRA induced apoptosis in the breast cancer cells. The MTT assay was used to determine drug-mediated cytotoxicity. A cell death detection ELISA kit was used for detection of the DNA fragments. The activity of serine/threonine protein phosphatases was assayed by the serine/threonine phosphatase system. The expression of serine/threonine protein phosphatases was evaluated by Western blot. During ATRA treatment, a significant decrease in the activity of serine/threonine phosphatases 2A, B and C occurred. The decreased activity of PP2A correlated with the up-regulation of PP2A catalytic and PP2A/B gamma, PP2A/B alpha regulatory subunits. The decrease in activity of the PP2B correlated with down-regulation of PP2B catalytic and up regulation of PP2B regulatory subunit expression. In addition, there was an up regulation in PP4C and down regulation in PP2C alpha/beta subunits protein expression. We demonstrated clear alteration in the activity and expression of serine/threonine protein phosphatases in breast cancer cells during ATRA treatment, and we suggest that the ATRA-induced apoptosis of the MCF-7 cells is significantly related to the phosphorylation dynamics. PMID- 14534749 TI - p53 gene mutations are rare in human papillomavirus-associated colon cancer. AB - Recent studies suggest that infection with high risk human papillomavirus (HPV) is a common event in colon tumors. Infection by oncogenic HPV may result in functional inactivation of the p53 protein in absence of mutations. Thus far no studies have been made to examine the frequency of p53 mutations in HPV associated colon cancer. The purpose of this study was to investigate the interrelationship between p53 mutations and HPV infection. The 'hot-spot' region of the p53 gene for mutations was analyzed by PCR-SSCP and direct sequencing in HPV-positive tumor samples. Only 2 mutations were identified in 56 samples. This rate was much lower than reported for sporadic colon tumors. Our results indicate an inverse relationship between p53 mutations and HPV infection and suggest that p53 inactivation caused by HPV infection may play a role in the pathogenesis of colon cancer. PMID- 14534750 TI - Genetic analysis of Agrobacterium tumefaciens susceptibility in Brassica oleracea. AB - The genetic control and heritability of Agrobacterium tumefaciens susceptibility was investigated using a doubled haploid (DH) mapping population of Brassica oleracea and the associated RFLP map. Preliminary studies were carried out by analysis of an 8 x 8 diallel, for which the parental lines were selected to include a range of susceptibilities to A. tumefaciens. The variation observed within the diallel was attributed to both additive and dominant gene effects, with additive gene effects being more important. A broad sense heritability value of 0.95 suggested that 95% of the observed variation was due to genetic effects, with just 5% attributed to non-genetic or environmental effects. A high narrow sense heritability value of 0.79 suggested that 79% of this trait was controlled by additive gene effects and, therefore, the potential to introduce this trait into breeding material is high. Fifty-nine DH lines from the mapping population were screened for susceptibility towards A. tumefaciens. Variation in susceptibility was observed across the population. The results of the DH screen were entered into the mapping programme MAPQTL and a highly significant quantitative trait loci (QTL) associated with susceptibility to A. tumefaciens was identified on linkage group 09. The use of substitution lines covering this region confirmed the location of this QTL. This work shows that susceptibility to A. tumefaciens is a heritable trait, and the transfer of susceptibility into resistant lines is demonstrated. These findings may help to overcome genotype restrictions to genetic transformation. PMID- 14534751 TI - Lr41, Lr39, and a leaf rust resistance gene from Aegilops cylindrica may be allelic and are located on wheat chromosome 2DS. AB - The leaf rust resistance gene Lr41 in wheat germplasm KS90WGRC10 and a resistance gene in wheat breeding line WX93D246-R-1 were transferred to Triticum aestivum from Aegilops tauschii and Ae. cylindrica, respectively. The leaf rust resistance gene in WX93D246-R-1 was located on wheat chromosome 2D by monosomic analysis. Molecular marker analysis of F(2) plants from non-critical crosses determined that this gene is 11.2 cM distal to marker Xgwm210 on the short arm of 2D. No susceptible plants were detected in a population of 300 F(2) plants from a cross between WX93D246-R-1 and TA 4186 ( Lr39), suggesting that the gene in WX93D246-R 1 is the same as, or closely linked to, Lr39. In addition, no susceptible plants were detected in a population of 180 F(2) plants from the cross between KS90WGRC10 and WX93D246-R-1. The resistance gene in KS90WGRC10, Lr41, was previously reported to be located on wheat chromosome 1D. In this study, no genetic association was found between Lr41 and 51 markers located on chromosome 1D. A population of 110 F(3 )lines from a cross between KS90WGRC10 and TAM 107 was evaluated with polymorphic SSR markers from chromosome 2D and marker Xgdm35 was found to be 1.9 cM proximal to Lr41. When evaluated with diverse isolates of Puccinia triticina, similar reactions were observed on WX93D246-R-1, KS90WGRC10, and TA 4186. The results of mapping, allelism, and race specificity test indicate that these germplasms likely have the same gene for resistance to leaf rust. PMID- 14534752 TI - Subungual melanoma with osteocartilaginous differentiation. AB - Osteocartilaginous metaplasia is known to occur rarely in melanomas, particularly in subungual melanomas. We present a case of a calcified subungual soft tissue tumour in which biopsy of the lesion showed malignant round and spindle-shaped tumour cells, many of which were associated with the formation of cartilage and osteoid-like material. Subsequent resection showed clear histological evidence of a subungual melanoma. Tumour cells expressed S100, melan-A and neurone-specific enolase but were negative for HMB45. Diagnostic radiological and histological features and the nature of the osteocartilaginous differentiation within this lesion is discussed. PMID- 14534753 TI - Analysis of peptaibol sequence composition: implications for in vivo synthesis and channel formation. AB - The sequence entries in the Peptaibol Database were analysed to provide information on compositional features of this unusual family of peptides. The non standard amino acid alpha-aminoisobutyric acid represents almost 40% of the residues in all the known sequences. Glutamine is the only significant polar residue in peptaibols, and the position and number of these residues appear to be related to their functional properties as ion channels. Aromatic residues are clustered at the termini, which may contribute to stabilization of the peptide vertically within the bilayer. The peptide chain length is strongly weighted towards the longer members of the family (16-20 residues) and likely to be an important feature in their mode of action as transmembrane permeabilizers. The significant skewing towards even numbers of residues and the bias in pairwise distributions of amino acids have implications for the nature of the in vivo synthesis of these peptides via large non-ribosomal protein complexes. PMID- 14534755 TI - Isovaleric acidaemia: cranial CT and MRI findings. AB - Isovaleric acidaemia is an inborn error of leucine metabolism due to deficiency of isovaleryl-CoA dehydrogenase, which results in accumulation of isovaleric acid in body fluids. There are acute and chronic-intermittent forms of the disease. We present the cranial CT and MRI findings of a 19-month-old girl with the chronic intermittent form of isovaleric acidaemia. She presented with severe metabolic acidosis, hyperglycaemia, glycosuria, ketonuria and acute encephalopathy. Cranial CT revealed bilateral hypodensity of the globi pallidi. MRI showed signal changes in the globi pallidi and corticospinal tracts of the mesencephalon, which were hypointense on T1-weighted and hyperintense on T2-weighted images. PMID- 14534756 TI - A fading Meckel's diverticulum: an unusual scintigraphic appearance in a child. AB - We describe the case of a 13-month-old boy with significant rectal bleeding in which the (99m)Tc pertechnetate scan showed an initial focus of uptake in the left iliac fossa, which faded rapidly at 15 min. At surgery an ulcerated Meckel's diverticulum was found. We therefore highlight the need to consider a Meckel's diverticulum in cases where this atypical scintigraphic appearance is seen. PMID- 14534754 TI - Intussusception. Part 3: Diagnosis and management of those with an identifiable or predisposing cause and those that reduce spontaneously. AB - In the previous two parts of this review on intussusception, the diagnosis and management of symptomatic, "idiopathic" ileocolic and ileoileocolic intussusceptions, which are considered to result from hyperplasia of lymphoid tissue in the distal ileum, were discussed. In this third part, those intussusceptions with an identifiable cause including pathologic lead point, those due to gastrojejunostomy or other feeding tubes, and those that are seen in the postoperative period as well as those that may be asymptomatic or may reduce spontaneously (usually limited to the small bowel) are discussed. PMID- 14534757 TI - Neuroimaging findings in glutaric aciduria type 1. AB - OBJECTIVE: To review the imaging features of glutaric aciduria type 1 (GA-1) in a group of 20 patients, the largest published series to date. To document the findings not previously reported and compare our findings with the imaging characteristics of GA-1 previously reported in the literature. MATERIALS AND METHODS: For 14 patients the original scans were examined and in the remaining 6, where the imaging was unavailable, the radiology reports were consulted. Nine patients had serial cranial US examinations, 13 had 18 CT scans performed and 14 patients had 39 MRI scans. RESULTS: Widening of the sylvian fissures and of the fluid spaces anterior to the temporal lobes was seen in 93% of cases. The mesencephalic cistern was also widened in 86%. Abnormal high-signal intensity on T2-weighted (T2-W) images was seen in the basal ganglia and periventricular white matter in 64% of children. Subdural collections were found in 3 patients, all of which resolved spontaneously. Four neonates followed with serial cranial US showed bilateral multiple caudothalamic cysts. Abnormal high signal on T2-W images was seen in the dentate nucleus, substantia nigra and the pontine medial lemniscus in 79, 43 and 64%, respectively. CONCLUSIONS: Widening of the sylvian fissure, mesencephalic cistern and expansion of CSF spaces anterior to the temporal lobes are cardinal signs of GA-1. If combined with abnormalities of the basal ganglia and white matter, GA-1 should be strongly suspected. PMID- 14534758 TI - Severe sinus bradycardia in a patient with Rett syndrome: a new cause for a pause? AB - Rett syndrome is a progressive disorder seen primarily in young females. It is characterized by autonomic dysfunction affecting many organ systems. Although sudden death is common in these patients, little is known about the cardiovascular manifestations of this dysautonomia. Earlier reports focused on the role of sympathetic overactivity manifest as reduced heart rate variability and prolonged QT intervals indicating a propensity for lethal ventricular arrhythmias. Bradyarrhthmias and sinus node dysfunction, however, have not previously been reported. We describe a 2-year-old with Rett syndrome and severe sinus bradycardia requiring a pacemaker implant. This case represents the first description of a new cardiovascular manifestation of Rett syndrome and may provide an explanation for sudden and unexpected death in some of these patients. PMID- 14534759 TI - Blalock-Taussig shunt for a neonatal case of Ebstein's anomaly with unobstructed right ventricular outflow tract. AB - A neonate with Ebstein's anomaly presented with severe cyanosis because of massive right-to-left shunt through an atrial septal defect and reduced blood flow through an unobstructed right ventricular outflow tract. This atypical patient underwent a right modified Blalock-Taussig shunt, which resolved the respiratory. PMID- 14534760 TI - Morbidities in patients with hypoplastic left heart syndrome. AB - We sought to document morbidities and growth for patients with hypoplastic left heart syndrome (HLHS) to inform the initial surgical decision and understand healthcare needs. Data were obtained on 137 patients with HLHS, born between 1989 and 1994, who survived staged surgery ( n = 62) or transplantation ( n = 75) and had follow-up information available from four pediatric cardiac surgical centers. In patients with HLHS older than 1 year of age at follow-up, 93% experienced at least one major postsurgical morbidity. Morbidities depended on the surgery received. Hypertension, renal compromise, and abnormal infections were more common in transplanted patients than staged surgery patients. Staged surgery patients used more anticongestive medications and experienced more morbidities requiring interventional catheterization than did transplanted patients. Rejection was common for transplanted patients. On average these children spent 23 days per year in the hospital. Patients with HLHS were small for their age; 43% of staged surgery patients weighed below the third percentile at last information, compared to 19% of transplanted patients ( p = 0.003). The median height percentile was the 10th in both groups. Normal activity level was reported in more transplanted patients (90%) than staged surgery patients (49%; p < 0.001). Trade-offs between mortality and morbidity outcomes can help inform the initial surgical decision. PMID- 14534761 TI - Intravenous nicardipine for treatment of postcoarctectomy hypertension in children. AB - Our objective was to evaluate the efficacy of intravenous (IV) nicardipine for the treatment of postcoarctectomy hypertension in children with coarctation of the aorta. We carried out a retrospective review in a pediatric intensive care unit at a tertiary care children's hospital. The patients were children with coarctation of the aorta treated for postcoarctectomy hypertension. Children with postcoarctectomy hypertension defined as a systolic blood pressure >95th percentile for age measured by indwelling arterial catheter were treated with IV nicardipine. We measured change in mean arterial blood pressure (MAP), mean systolic and diastolic blood pressure, and mean heart rate (HR) from baseline after initiating treatment with IV nicardipine. The outcome measure was a reduction in MAP and mean systolic and diastolic blood pressure after treatment with IV nicardipine. During a 4-year period, 10 children met the study criteria. Median age was 3.25 months (range, 0.25 to 180 months). Initial median treatment dose of IV nicardipine was 1.0 micro g/kg/min (range, 0.5 to 6 micro g/kg/min); median dose used to control hypertension was 1.5 micro g/kg/min (range, 0.25 to 6 micro g/kg/min). Median duration of therapy was 26.3 h (range, 13 to 49 h). Treatment with IV nicardipine resulted in a 26.5% decrease in MAP from baseline during the first hour of treatment ( p = 0.0006). Mean systolic blood pressure decreased from 133 to 105 mmHg ( p = 0.005), and mean diastolic blood pressure decreased from 75 to 52.5 mmHg ( p = 0.001) during the first hour of therapy with nicardipine. There was a significant reduction ( p = 0.0005) in MAP during continued treatment with IV nicardipine. The mean HR of 150 remained unchanged during the first hour of therapy with nicardipine, and no significant change in mean HR or adverse effects was noted during continued therapy. Two children receiving other antihypertensive therapy demonstrated further reduction in their blood pressure when IV nicardipine was initiated. Tachycardia and hypotension were not observed in any child treated with IV nicardipine. We concluded that IV nicardipine reduced MAP with no significant change in mean HR and no adverse effects in patients with postcoarctectomy hypertension. Nicardipine produced a further reduction in MAP in children receiving other antihypertensive agents. Nicardipine is an effective agent for treatment of postcoarctectomy hypertension in children with coarctation of the aorta. PMID- 14534762 TI - Resolution of plastic bronchitis with atrial pacing in a patient with fontan physiology. AB - We describe a 5-year-old boy with Fontan physiology and a ventricular pacemaker who developed severe plastic bronchitis. Evaluation by cardiac catheterization revealed profoundly altered hemodynamics, which improved with atrial pacing. Following implantation of an atrial pacemaker, which restored atrioventricular (A V) synchrony, the patient's hemodynamics greatly improved and his plastic bronchitis resolved. PMID- 14534763 TI - Evaluation of the efficacy of treatment of Kawasaki disease before day 5 of illness. AB - We evaluated the efficacy of treating Kawasaki disease earlier than Day 5 of illness with a standard dose of immunoglobulin and aspirin. We performed a case control study of patients with Kawasaki disease admitted to Princess Margaret Hospital from 1994 to 1999. Patients with pretreatment coronary aneurysm or those treated after day 10 of illness were excluded. All patients received immunoglobulin (2 g/kg) and aspirin (80-100 mg/kg/day) until fever subsided for 48 hours. Immunoglobulin retreatment was given for persistent fever 48 hours after the first dose of immunoglobulin or recrudescent fever. The case group consisted of 15 patients who received treatment earlier than day 5 of illness, and the control group consisted of 66 patients who were treated on or after day 5. Patients' sex, age, duration of posttreatment fever, need for additional immunoglobulin, and coronary artery status were noted. Treatment efficacy was assessed by the duration of posttreatment fever and the prevalence of coronary artery aneurysms. Eighty-one patients were included in this study. There were 15 patients in the case group and 66 in the control group. No significant difference was noted in age and sex between the case and control groups. Thirty-three percent (5/15) and 8% (5/66) of the case and control groups, respectively, had persistent/ recrudescent fever 48 hours after the first dose of immunoglobulin that required retreatment ( p = 0.017). Thirteen percent (2/15) and 5% (3/66) of the case and control groups, respectively, had coronary aneurysms ( p = 0.158). Treatment of Kawasaki disease before day 5 of illness was associated with persistent/recrudescent fever that required retreatment. However, there was no significant increase in the prevalence of coronary aneurysm if retreatment was given. PMID- 14534764 TI - Proton magnetic resonance spectroscopy with a body coil in the diagnosis of carcinoma prostate. AB - We evaluate the feasibility of proton magnetic resonance spectroscopy ((1)H MRS) using a spine coil receiver (body coil) in the diagnosis of carcinoma prostate. Seventeen patients with biopsy-proven prostate carcinoma, five patients of benign prostatic hyperplasia (BPH) and five healthy young volunteers underwent (1)H MRS investigation. MRS was performed at 1.5 Tesla using a spine receiver coil for signal reception. In vivo citrate levels are reported as a ratio of citrate peak area to the sum of the areas of choline and creatine peak. MRS spectrum with good sensitivity and signal to noise (S/N) ratio was obtained in all 27 subjects. The citrate to creatine plus choline ratio was 0.31+/-0.25 in patients with cancer, 1.43+/-0.58 in BPH and 2.16+/-0.56 in controls. The difference in ratios between cancer and BPH and cancer and control was statistically significant ( p<0.01). Within the cancer group, there was a statistically significant decline in levels with higher-grade malignancy ( p<0.05). There were no complications of the procedure. There is a statistically significant decline in the ratio of citrate to choline plus creatine in the regions of cancer prostate when compared with BPH or normal control. The study demonstrates that MRS data can be reliably acquired using a spine coil receiver. MRS may also play a role in differentiating well differentiated tumors from the anaplastic variety. PMID- 14534766 TI - Navigated diffusion-weighted imaging with interpolated phase-correction for high resolution imaging of stroke. AB - Stroke imaging was revolutionised with the introduction of diffusion-weighted MRI (DWI). The commonly used echoplanar DWI suffers from geometrical distortion near the skull base and the frontal regions and from reduced spatial resolution and fat suppression. To allow a voxel-by-voxel comparison between high-resolution spin-echo images, we implemented spin-echo-based DWI. Motion artefacts were eliminated by phase correction in hybrid frequency-Fourier domain using navigator echoes. In a novel approach, distorted navigator echoes which did not eliminate motion artefacts were replaced with interpolated navigator echoes, leading to restored image information. Navigated DWI yielded high-resolution images in 21 of 24 patients with brain ischaemia, allowing diagnosis of even small or diffuse zones of ischaemia. We determined the spatial distribution and mean of T(2)- and DWI signal intensity and apparent diffusion coefficients (ADC), using a multidimensional histogram-based analysis. Mean ADC were decreased in ischaemic areas less than 9 days old. The technique may also be useful for high-resolution DWI of tissue other than the brain. PMID- 14534765 TI - The evaluation of rat prostate blood flow using a laser speckle flowmetry: a comparative study using the microsphere method in castrated and androgen replenished rats. AB - To evaluate the accuracy and reliability of laser speckle blood flowmetry in the measurement of prostate blood flow, we performed a comparative study with the ordinary intra-arterial radioactive microsphere injection method using the well known castrate-rat model. Adult male Wistar strain rats were used for the study. The rats were either only castrated or subcutaneous testosterone replenishment was followed subcutaneously 6 h after castration. The prostate blood flow was measured at different time courses after castration. The effect of a single androgen replenishment on prostate blood flow was also evaluated. The measurement of prostate blood flow was performed by both the radioactive microsphere injection method and laser speckle blood flowmetry, and then the outcomes were compared. Rapid reduction in prostate blood flow to 30% of the normal level was observed 24 h after castration in the measurements with the microsphere injection technique. The prostate blood flow decreased gradually after 24 h post-castration to 23 and 21% of the normal level at 48 and 72 h after castration, respectively. The laser speckle blood flowmetry also detected the decrease in prostate blood flow well, but in a more gradual manner. The prostate blood flow was 70, 52 and 35% of the normal level at 24, 48 and 72 h after castration, respectively. The effect of a single administration of testosterone to castrated rats had recovered the prostate blood flow to 74 and 98% of the normal level by measurement with the microsphere injection technique and laser speckle blood flowmetry, respectively. The different outcome in blood flow rate change between the methods can be explained according to their different mechanism of measurement, thus suggesting the capillary vessels are the early and most responsive portion for hormonal manipulation. In conclusion, the laser speckle blood flowmetry is a convenient and reliable method for evaluating prostate blood flow, especially when the organ is required for other biological and molecular assays, since the method does not require the excision of the organ for the measurement. PMID- 14534767 TI - Rupture of an internal carotid artery aneurysm during angiography with leakage of contrast medium via an external ventricular drain. AB - With a ruptured intracranial aneurysm producing subarachnoid haemorrhage (SAH) cerebral angiography is currently used for identification of the affected vessel. Aneurysm rerupturing is one of the more serious complications of cerebral angiography and has been frequently described. We report a 61-year-old man who presented with SAH who had rerupture of a large aneurysm of the internal carotid artery during angiography. A substantial amount of contrast medium escaped via a ventricular drain. The three main risk factors for rerupture of an aneurysm are: angiography performed within 6 h of the primary SAH, an aneurysm on the internal carotid artery and an unfavourable Glasgow coma score. PMID- 14534768 TI - Follow-up of intracranial aneurysms treated with detachable coils: comparison of plain radiographs, 3D time-of-flight MRA and digital subtraction angiography. AB - All patients with aneurysms treated with Guglielmi detachable coils (GDC) are undergo angiography to assess long-term stability of aneurysm exclusion or to show recurrence of the aneurysm sac, which may require further treatment. We prospectively compared the plain-film appearance of the coil-mass, 3D time-of flight MR angiography (TOF MRA) and digital subtraction angiography (DSA) for the detection of aneurysm recanalisation during follow-up. We studied 60 patients with 74 intracranial aneurysms treated with Guglielmi detachable coils. We used the unsubtracted image of the angiograms performed at the completion of any embolisation procedure and at follow-up as the plain radiographs. Recanalisation was considered if loosening, compaction or reorientation of the coil mass was apparent. TOF MRA was performed to assess the presence and size of a neck remnant. DSA was regarded as the definitive investigation. Comparison of the techniques showed good agreement as regards aneurysm recanalisation. MRA was more accurate than plain radiography and could replace DSA for long term follow- up. The initial follow-up examination should, however, include both modalities. In cases of contraindications or limitations to MRA, the interval between follow-up angiographic examinations could be increased if there is no change in the plain film coil-mass appearances. PMID- 14534769 TI - Modeling variability in cortical representations of human complex sound perception. AB - This study investigated methodological (task, stimulus) and intersubject variability in the cortical representation of auditory processing of complex sounds, including speech. Subjects were adult seizure patients undergoing left hemisphere electrocortical mapping (ECM). We tested auditory discrimination of complex sounds, including frequency-modulated tones and speech syllables (digitized, synthesized) contrasted by phonetic features and lexical status. To measure task effects, auditory comprehension was also tested. Within- and across patient differences in the distribution of deficits induced by ECM were modeled statistically using the recently developed method of Template Mixture Modeling. Cortical representations of auditory discrimination were smaller, more localized, and less variable across subjects than auditory comprehension. Stimulus effects were observed only for speech-tone contrasts. When tasks and stimuli were held constant, two auditory discrimination centers were identified in the posterior temporal lobe. There was also an interaction between task and intersubject effects, with more intersubject variability in cortical maps of auditory comprehension than auditory discrimination. These results demonstrate the utility of using the statistical modeling approach of Template Mixture Modeling to quantify sources of variability in cortical functional organization. PMID- 14534770 TI - Changes in corticospinal motor excitability induced by non-motor linguistic tasks. AB - The excitability of the corticospinal motor pathways to transcranial magnetic stimulation (TMS) can be differentially modulated by a variety of motor tasks. However, there is emerging evidence that linguistic tasks may alter excitability of the corticospinal motor pathways also. In this study we evaluated the effect of several movement-free, low-level linguistic processes involved in reading and writing on the excitability of the bilateral corticospinal motor pathways in a group of right-handed subjects. The study included two series of tasks, visual searching/matching and imaginal writing/drawing. The tasks were designed to roughly correspond with elemental aspects of the reading and writing, grapheme recognition and grapheme generation, respectively. Each task series included separate blocks with different task targets: letters, digits, semantically easy to-code (i.e. geometric) shapes, and semantically hard-to-code shapes, as well as control blocks with no task. During task performance, TMS was delivered randomly over the hand area of either the left or right motor cortex and the modulation of the excitability of the corticospinal motor pathways was measured bilaterally through changes of the size of the motor-evoked potential (MEP) induced in the relaxed right and left first dorsal interosseous (FDI) muscles. We found that the size of the MEP in hand muscles increased during visual searching/matching tasks, particularly when targets were letters or geometric shapes, and the increase was significant for the dominant hand (left hemisphere) only. No such consistent effects were seen across subjects during imaginal tasks. This study provides evidence that even the performance of certain low-level linguistic tasks can modulate the excitability of the corticospinal motor pathways, particularly those originating from the left (dominant) hemisphere, despite the absence of overt motor activity. Moreover, in the light of the recently increased awareness of the role of "mirror neurons" in perception, the results suggest that activation of motor circuits used in generation of the written output may be an essential part of the perception of the written material as well. Understanding the patterns of task-dependent changes in excitability of the corticospinal motor pathways will provide insights into the organisation of central nervous system functional networks involved in linguistic processes, and may also be useful for future development of novel approaches to rehabilitation therapy of linguistic and motor functions. PMID- 14534771 TI - Chronic transdermal nicotine patch treatment effects on cognitive performance in age-associated memory impairment. AB - OBJECTIVES: Chronic transdermal nicotine has been found to improve attentional performance in patients with Alzheimer's disease (AD), but little is known about chronic nicotine effects in age-associated memory impairment (AAMI), a milder form of cognitive dysfunction. The current study was performed to determine the clinical and neuropsychological effects of chronic transdermal nicotine in AAMI subjects over a 4-week period. DESIGN: The double-blind, placebo-controlled, cross-over study consisted of two 4-week periods separated by a 2-week washout period. SETTING: An outpatient setting was used. PARTICIPANTS: The subjects ( n=11) met criteria for AAMI. INTERVENTIONS: The subjects were given nicotine patches (Nicotrol) to wear for 16 h a day at the following doses: 5 mg/day during week 1, 10 mg/day during week 2 and week 3 and 5 mg/day during week 4. MEASUREMENTS: The effects of nicotine treatment were determined with the clinical global impressions questionnaire, Conners' Continuous Performance test, and the automated neuropsychologic assessment metrics (ANAM) computerized neuropsychology battery. RESULTS: Nicotine significantly improved the clinical global impression score as assessed by participants, as well as objective tests of attentional function on the Connors' Continuous Performance Test and decision reaction time on the neuropsychology test battery. Nicotine did not improve performance on other tests measuring motor and memory function. CONCLUSION: Chronic transdermal nicotine treatment in AAMI subjects caused a sustained improvement in clinical symptoms and objective computerized tests of attention. These results support the further investigation of nicotinic treatment as a promising therapy for AAMI. PMID- 14534774 TI - Diagnosis and treatment of antibody-mediated kidney allograft rejection. AB - Evidence of a significant pathogenetic role of donor-reactive antibodies (DRA) in kidney allograft rejection is accumulating. At least, partially owing to the recent discovery of the complement split product C4d as a valuable rejection marker, antibody-mediated rejection (AMR) has regained increasing attention. We review here the value of various diagnostic criteria, including immunohistochemistry (C4d staining), histomorphology and posttransplant serology, for the diagnosis of AMR. Furthermore, the mechanisms underlying alloantibody/complement-mediated allograft injury are discussed in detail. Finally, a thorough discussion of recently proposed "anti-humoral" therapeutic strategies is provided. PMID- 14534772 TI - Differential region-specific regulation of central alpha 1-adrenoceptor binding following chronic haloperidol and clozapine administration in the rat. AB - RATIONALE: Many antipsychotics exhibit potent anti-alpha(1)-adrenergic receptor activity, which has been suggested to contribute to typical and atypical antipsychotic effects and to the production of centrally mediated side effects. OBJECTIVES: To assess the relative contribution of alpha(1)-adrenoceptors to the mechanism of action of haloperidol and clozapine and to identify possible sites of action. METHODS: We examined the effect of chronic haloperidol and clozapine treatment on alpha(1)-adrenoceptor characteristics in several rat brain regions. For comparison, D(2)-like dopamine receptor density in the striatum was also determined. RESULTS: Clozapine administration (25 mg/kg/day i.p., 21 days) significantly increased alpha(1)-adrenoceptor density in the frontal cortex (44%), remaining cortex (49%) and thalamus (93%) but binding levels in the hippocampus and spinal cord were unchanged relative to vehicle. Haloperidol treatment (1.5 mg/kg/day i.p., 21 days) also significantly increased the density of alpha(1)-adrenoceptor binding in the thalamus (73%), but had no effect on alpha(1)-adrenoceptor levels in any other region examined. alpha(1)-Adrenoceptor affinity in the cortex was not significantly altered by either antipsychotic treatment. Haloperidol, in contrast to clozapine, significantly upregulated dopamine D(2)-like binding in the striatum. CONCLUSIONS: Central alpha(1) adrenoceptors are differentially regulated after chronic haloperidol and clozapine treatment. It is suggested that thalamic alpha(1)-adrenoceptors may represent a common anatomical locus contributing to the antipsychotic activity and/or alpha(1)-adrenoceptor centrally mediated side effects of both drugs, whereas the selective upregulation of cortical alpha(1)-adrenoceptor density by clozapine may contribute, in part, to its superior atypical properties. PMID- 14534773 TI - Predicting the effects of estrogen replacement therapy on lumbar bone mineral density in oophorectomized women: analysis of a 10-year longitudinal study. AB - The purpose of this study was to investigate whether it is possible to predict the long-term effects of estrogen replacement therapy (ERT) on lumbar bone mineral density (BMD) of oophorectomized women based on changes in BMD. In this study, we retrospectively investigated the changes in lumbar BMD of 70 oophorectomized women under ERT for more than 10 years, and examined whether it was possible in the early stage of ERT to predict the amount of lumbar BMD based on various parameters. Seventy oophorectomized Japanese women (56.8+/-3.9 years old) treated with conjugated equine estrogen (oral) at a dosage of 0.625 mg/day for 10 years were enrolled. Lumbar (L2-L4) BMD was measured annually by dual energy X-ray absorptiometry (DXA; CV<1.0%). The correlation between changes in BMD after 10 years on ERT (DeltaBMD10) and several clinical factors was examined using a stepwise multiple regression model. The change in BMD after 1 year on ERT (%DeltaBMD1) was the only independent factor that correlated with changes in BMD after 10 years on ERT; the coefficient of correlation was R(2)=0.557 ( R=0.750, P<0.001). Based on the %DeltaBMD1, the 70 women were divided into two groups: women with a positive change in the 1st year (%DeltaBMD1 >or=0%) in group A ( n=40) and those with a negative value in the first year ( %DeltaBMD1<0%) in group B ( n=30). We investigated the sensitivity and specificity in the coincidence of %DeltaBMD1 changes in BMD after 10 years on ERT. The %DeltaBMD1 coincided with changes in BMD after 10 years on ERT; the sensitivity was 92.5% and specificity was 70.0%. In conclusion, changes in lumbar BMD on ERT can be predicted from the changes in lumbar BMD at the end of the 1st year. PMID- 14534775 TI - Differentiated HFO and CMV for independent lung ventilation in a pediatric patient. PMID- 14534776 TI - Gastric PgCO2 and Pg-aCO2 gap are related to D-lactate and not to L-lactate levels in patients with septic shock. AB - OBJECTIVE: Intestinal ischemia causes an increase in lactate production and gastric intramucosal carbon dioxide partial pressure (PgCO(2)). However, no linear relationship between systemic l-lactate levels and gastric tonometry during intestinal ischemia has been found, probably since l-lactate is rapidly cleared from the circulation by the liver. In contrast, the rate of d-lactate clearance from the circulation by the liver is considerably lower than that of l lactate, and d-lactate may therefore be more closely related to measurements of gastric tonometry than l-lactate values. DESIGN AND SETTING: Prospective, observational study in a university-affiliated mixed intensive care unit. SUBJECTS: Twenty critically ill patients with septic shock. MEASUREMENTS AND RESULTS: During the first 24 h of admission to the intensive care unit at least two blood samples were taken for d- and l-lactate measurements and arterial blood gases, Simultaneously, gastric PgCO(2) was measured using capnographic tonometry. The intramucosal-arterial PCO(2) gap was calculated using gastric intramucosal PgCO(2) and arterial PCO(2) from arterial blood. d-Lactate was significantly correlated to PgCO(2) values and to the mucosal-arterial PCO(2) gap. There was no relationship between l-lactate and PgCO(2) or the mucosal-arterial PCO(2) gap. d lactate and l-lactate values were significantly correlated. CONCLUSIONS: During sepsis intestinal production of d-lactate is related to gastric intramucosal PCO(2). No such relationship was found between l-lactate values and PgCO(2) PMID- 14534777 TI - Using hierarchical modeling to measure ICU quality. AB - OBJECTIVE: To determine whether hierarchical modeling agrees with conventional logistic regression modeling on the identity of ICU quality outliers within a large multi-institutional database. DESIGN: Retrospective database analysis. SETTING AND PATIENTS: Subset of the Project IMPACT database consisting of 40435 adult patients admitted to surgical, medical, and mixed surgical-medical ICUs ( n=55) between 1997 and 1999 who met inclusion criteria for SAPS II. MEASUREMENTS AND RESULTS: The SAPS II score was customized to this database using conventional logistic regression and using a hierarchical (random coefficients) model. Both models exhibited excellent discrimination ( Cstatistic) and calibration (Hosmer Lemeshow statistic). The hierarchical and nonhierarchical models had C statistics of.870 and.865, and HL statistics of 3.71 ( p>.88, df=8) and 8.94 ( p>.35, df=8), respectively. Since the random effects component of the hierarchical model accounts for between-hospital variability, only the fixed-effects coefficients were used to calculate the expected mortality rate based on the hierarchical model. The ratio and 95% confidence intervals of the observed to expected mortality rate were calculated using both models for each ICU. ICUs whose observed/expected ratio was either less than 1 or greater than 1, and whose 95% confidence interval did not include 1 were labeled as either high-performance or low-performance outliers, respectively. Analysis using kappa statistic revealed almost perfect agreement between the two models (nonhierarchical vs. hierarchical) on the identity of ICU quality outliers. CONCLUSIONS: Models obtained by customizing SAPS II using a nonhierarchical and a hierarchical approach exhibit excellent agreement on the identity of ICU quality outliers. PMID- 14534779 TI - Parenteral nutrition in the critically ill. PMID- 14534781 TI - Founder population size and number of source populations enhance colonization success in waterstriders. AB - Understanding the factors that underlie colonization success is crucial both for ecological theory and conservation practices. The most effective way to assess colonization ability is to introduce experimentally different sets of individuals in empty patches of suitable habitat and to monitor the outcome. We translocated mated female waterstriders, Aquarius najas, into 90 streams that were not currently inhabited by the species. We manipulated sizes of propagules (from 2 to 16 mated females) and numbers of origin populations (one or two). Three origin populations were genetically different from each other, but they were less than 150 km from the streams of translocation. The results demonstrate clearly that both the larger propagule size and the high number of source populations have positive effects on the probability of colonizing a new stream. Thus, in addition to the stochastic factors related to the propagule size it may be essential to consider also the diversity of genetic origin for colonization success. PMID- 14534780 TI - Elevated serum vascular endothelial growth factor is associated with visceral fat accumulation in human obese subjects. AB - AIMS/HYPOTHESIS: Adipose tissue expresses some bioactive molecules, which may be involved in the development of obesity-associated metabolic disorders and cardiovascular diseases. Vascular endothelial growth factor (VEGF) an important angiogenic factor is implicated in normal and pathological vessel formation. The aim of this study is to investigate clinically the association between blood serum VEGF concentrations and body fat accumulation as well as distribution. The study also aims to show the effect of serum VEGF protein on gene expression of transcriptional factor E26 transformation-specific-1 (Ets-1) and matrix metalloproteinase (MMP)-3. METHODS: Serum VEGF concentrations were measured in 38 overweight or obese subjects. Fat distribution in the abdominal subcutaneous as well as visceral fat areas was assessed by computed tomography scans at umbilical level. Furthermore, the changes of serum VEGF concentrations following body weight reduction therapy were analyzed in eight subjects recruited from the original pool of subjects. Semi-purified circulating VEGF proteins were obtained by heparin-sepharose and its biological activities were shown to alter gene expressions in human aortic endothelial cells. RESULTS: Serum VEGF concentrations were positively correlated with BMI (r=0.433, p=0.007) and visceral fat area (r=0.488, p=0.002). Stepwise regression analysis showed the visceral fat area as the most important determinant factor for VEGF circulating levels. Following body weight reduction therapy, VEGF concentrations as well as visceral fat area were decreased. The serum semi-purified VEGF protein enhanced expressions of Ets-1 and MMP-3 in human aortic endothelial cells. CONCLUSION/INTERPRETATION: Increased serum VEGF concentrations associated with visceral fat accumulation could influence vascular endothelial function. PMID- 14534782 TI - Effectiveness of six species of rodents as dispersers of singleleaf pinon pine (Pinus monophylla). AB - The effectiveness of six species of rodents as dispersers of singleleaf pinon pine (Pinus monophylla) was investigated in field enclosures. The rodent species were Panamint kangaroo rat (Dipodomys panamintinus), Great Basin pocket mouse (Perognathus parvus), pinyon mouse (Peromyscus truei), deer mouse (Peromyscus maniculatus), least chipmunk (Tamias minimus), and white-tailed antelope ground squirrel (Ammospermophilus leucurus). Five measures of seed harvesting and caching behavior were assessed and used to calculate the effect of species on the probability of effective dispersal (i.e., movement of seeds to potential establishment sites) of pinon pine seeds. These measures were proportion of seeds harvested, proportion of harvested seeds that were scatter hoarded, relative dispersion of scatter-hoarded seeds, seedling emergence and seedling establishment. To evaluate the effects of the last two variables, a 2-year study of seed germination and seedling establishment was conducted in the field. Seedling survival was 72-90% for seedlings emerging from 1-4 cm deep but was markedly lower from surface caches and for depths >4 cm. All seedlings in the open microsite perished before the end of the first year, whereas 38-97% of seedlings under shrubs survived. Cache size did not affect seedling survival over the 2 years, but large caches reduced the total number of potential establishment sites. All six species harvested over 90% of the pinon seeds presented and scatter hoarded from 31% (pocket mouse) to 66% (chipmunk) of the seeds they harvested. Pinon mice and deer mice distributed seeds to more cache sites (i.e., put fewer seeds in caches) than the other species and, consequently, created more potential establishment sites. All species scatter hoarded seeds at depths suitable for seedling emergence, but pocket mice and pinon mice cached under shrubs more than the other species, the microhabitat where seedlings had the greatest probability of establishing. Factors that had the greatest effect on the probability of effective dispersal were microhabitat selection and cache size. PMID- 14534783 TI - Cis-acting elements sufficient for induction of FDH1 expression by formate in the methylotrophic yeast Candida boidinii. AB - The FDH1 gene of Candida boidinii encodes an NAD+-dependent formate dehydrogenase, which catalyzes the last reaction in the methanol dissimilation pathway. FDH1 expression is strongly induced by methanol, as are the promoters of the genes AOD1 (alcohol oxidase) and DAS1 (dihydroxyacetone synthase). FDH1 expression can be induced by formate when cells are grown on a medium containing glucose as a carbon source, whereas expression of AOD1 and DAS1 is completely repressed in the presence of glucose. Using deletion analyses, we identified two cis-acting regulatory elements, termed UAS-FM and UAS-M, respectively, in the 5' non-coding region of the FDH1 gene. Both elements were necessary for full induction of the FDH1 promoter by methanol, while only the UAS-FM element was required for full induction by formate. Irrespective of whether induction was achieved with methanol or formate, the UAS-FM element enhanced the level of induction of the FDH1 promoter in a manner dependent on the number of copies, but independent of their orientation, and also converted the ACT1 promoter from a constitutive into an inducible element. Our results not only provide a powerful promoter for heterologous gene expression, but also yield insights into the mechanism of regulation of FDH1 expression at the molecular level. PMID- 14534784 TI - Tuberculous peritonitis mimicking peritonitis carcinomatosis: a case report. AB - A 13-year-old girl presented with non-specific symptoms of peritonitis, low grade fever and missed her normal menses. Computed tomography of her abdomen and pelvic cavity showed omental cake and a generalised thickened peritoneum. An elevated serum cancer antigen-125 (CA-125) level of 1248.5 U/ml was also noted and a diagnosis of peritonitis carcinomatosis was first suspected. Diagnostic laparoscopy revealed multiple tubercles over the whole peritoneum and the pathology report described granulomatous nodules with giant cells and epithelioid cells. Culture of the ascitic fluid revealed Mycobacterium tuberculosiswhich resulted in a final diagnosis of tuberculous peritonitis. The patient's fever and abdominal distension gradually subsided after anti-tuberculosis treatment. The serum CA-125 level also decreased significantly to 10.2 U/ml after treatment. CONCLUSION: cancer antigen-125 levels may serve as a potential follow-up marker of disease activity and treatment response in tuberculous peritonitis. PMID- 14534785 TI - Reproducibility of the 1998 World Health Organization/International Society of Urologic Pathology classification of papillary urothelial neoplasms of the urinary bladder. AB - OBJECTIVES: This study assessed the diagnostic agreement and intra- and inter observer reproducibility of the World Health Organization/International Society of Urologic Pathology Consensus Classification of Urothelial Neoplasms (1998 WHO/ISUP classification) and the 1973 WHO classification. METHODS: A teaching set with 5 slides of each papillary neoplasm of low malignant potential, low-grade papillary carcinoma, high-grade papillary carcinoma, and a guideline, as well as a study set of 30 slides containing ten cases of each category, were sent to participants. Six pathologists expert in urological pathology reviewed the 30 slides of non-invasive papillary urothelial tumors in the study set. Diagnostic accuracy and reproducibility were evaluated using intra- and inter-rater techniques (kappa statistic). RESULTS: A moderate to substantial intra- and inter observer reproducibility was achieved for both the 1998 WHO/ISUP and 1973 WHO classification. The results of the two classification systems were not different statistically ( P>0.05). Reproducibility was lower in low-grade tumors for both classifications. CONCLUSIONS: The new proposed classification system for non invasive urothelial neoplasms does not increase the reproducibility. There is still a need for uniformity in grading in order to compare the different studies and therapies and to provide more accurate information for management. PMID- 14534786 TI - The cellular localization of prosystemin: a functional role for phloem parenchyma in systemic wound signaling. AB - The systemin precursor, prosystemin, has been previously shown to be sequestered in vascular bundles of tomato ( Lycopersicon esculentum Mill.) plants, but its subcellular compartmentalization and association with a specific cell type has not been established. We present in situ hybridization and immunocytochemical evidence at the light, confocal, and transmission electron microscopy levels that wound-induced and methyl jasmonate-induced prosystemin mRNA and protein are exclusively found in vascular phloem parenchyma cells of minor veins and midribs of leaves, and in the bicollateral phloem bundles of petioles and stems of tomato. Prosystemin protein was also found constitutively in parenchyma cells of various floral organs, including sepals, petals and anthers. At the subcellular level, prosystemin was found compartmentalized in the cytosol and the nucleus of vascular parenchyma cells. The cumulative data indicate that vascular phloem parenchyma cells are the sites for the synthesis and processing of prosystemin as a first line of defense signaling in response to herbivore and pathogen attacks. PMID- 14534787 TI - Transforming petals into sepaloid organs in Arabidopsis and oilseed rape: implementation of the hairpin RNA-mediated gene silencing technology in an organ specific manner. AB - Oilseed rape ( Brassica napus L.) genotypes with no or small petals are thought to have advantages in photosynthetic activity. The flowers of field-grown oilseed rape form a bright-yellow canopy that reflects and absorbs nearly 60% of the photosynthetically active radiation (PAR), causing a severe yield penalty. Reducing the size of the petals and/or removing the reflecting colour will improve the transmission of PAR to the leaves and is expected to increase the crop productivity. In this study the 'hairpin' RNA-mediated (hpRNA) gene silencing technology was implemented in Arabidopsis thaliana (L.) Heynh. and B. napus to silence B-type MADS-box floral organ identity genes in a second-whorl specific manner. In Arabidopsis, silencing of B-type MADS-box genes was obtained by expressing B. napus APETALA3( BAP3) or PISTILLATA ( BPI) homologous self complementary hpRNA constructs under control of the Arabidopsis A-type MADS-box gene APETALA1 ( AP1) promoter. In B. napus, silencing of the BPI gene family was achieved by expressing a similar hpRNA construct as used in Arabidopsis under the control of a chimeric promoter consisting of a modified petal-specific Arabidopsis AP3 promoter fragment fused to the AP1 promoter. In this way, transgenic plants were generated producing male fertile flowers in which the petals were converted into sepals ( Arabidopsis) or into sepaloid petals ( B. napus). These novel flower phenotypes were stable and heritable in both species. PMID- 14534788 TI - Identification of a new gene controlling plant height in rice using the candidate gene strategy. AB - A gene underlying a quantitative trait locus (QTL) controlling plant height on chromosome 1 (QTLph1) in rice ( Oryza sativa L.) was identified using the candidate-gene strategy. First, the function of a targeted gene was analyzed using near isogenic lines (NILs) in which the chromosomal region of a targeted QTL was substituted with that of another line. Second, for physiological information, the candidate gene was selected in the annotation data by the genome sequencing. Physiological analyses of an NIL-expressing QTLph1 (NIL6) suggested that the targeted gene controls plant height by enabling higher amounts of sucrose to be translocated in leaves. The results indicated that the gene for sucrose phosphate synthase (SPS; EC 2.4.1.14), the major limiting enzyme for sucrose synthesis, is a candidate gene for QTLph1 among the annotation results of the region of QTLph1. The higher level of SPS transcripts and the activity of SPS in NIL6 compared to control plants, and the fact that the relative SPS activity per SPS protein content was almost the same between NIL6 and Nipponbare suggested that the higher plant height in NIL6 compared to Nipponbare was due to the high SPS activity in NIL6. In agreement with this hypothesis, transgenic rice plants with a maize SPS gene that had about 3 times the SPS activity of that in Nipponbare (control plants) were significantly taller than Nipponbare from the early growth stage. From these results and the physiological data from NIL6, we concluded that SPS is the targeted gene underlying QTLph1. PMID- 14534789 TI - Post-translational modifications of alpha-tubulin in Zea mays L are highly tissue specific. AB - To further understand post-translational modifications (PTMs) of plant alpha tubulin, post-translationally modified alpha-tubulin isoforms from selected tissues of Zea mays L. were examined using two-dimensional electrophoresis and immunoblotting. Except for polyglycylated tubulin, tyrosinated, detyrosinated, acetylated and polyglutamylated alpha-tubulin isoforms were all present in maize tissues. Tyrosinated alpha-tubulin was the predominant variant in all cases, with isoforms alpha1-alpha4 (alpha5) being the most common components. Leaves exhibited a striking difference in PTM patterns of alpha-tubulin isoforms compared to other tissues examined. In leaves, several major specific isoforms were highly modified by detyrosination, acetylation and polyglutamylation. In pollen and anthers, only the most abundant isoform alpha3 was acetylated to an appreciable extent, and no acetylated isoform was found in roots. Similarly, in pollen, anthers and roots, only alpha3 was appreciably polyglutamylated. Additionally, a detyrosinated isoform alpha6 was present in anthers and in leaves, while the tyrosinated isoform alpha6 seemed to be pollen specific. These results indicate that certain types of PTM of plant alpha-tubulin preferentially occur in a tissue-specific way. PMID- 14534790 TI - Angiotensin-(1-7) stimulates water transport in rat inner medullary collecting duct: evidence for involvement of vasopressin V2 receptors. AB - The peptide angiotensin-(1-7) [Ang-(1-7)] is known to enhance water transport in rat inner medullary collecting duct (IMCD). The aim of this study was to determine the mechanism of the Ang-(1-7) effect on osmotic water permeability (Pf). Pf was measured in the normal rat IMCD perfused in vitro in presence of agonists [Ang-(1-7), arginine vasopressin (AVP) and Ang-(3-8)], and antagonists of the angiotensin and the vasopressin cascade. Ang-(1-7), but not Ang-(3-8), increased Pf significantly. The effect of Ang-(1-7) on Pf was abolished by its selective antagonist, A-779, added before or after Ang-(1-7). Prostaglandin E2 and the protein kinase A inhibitor H8 also blocked the Ang-(1-7) effect. Blockade of vasopressin V1 receptors by antagonists did not change the Ang-(1-7) effect, but pre-treatment with a V2 antagonist abolished the effect of Ang-(1-7) on Pf. Similarly, pre-treatment with A-779 inhibited AVP's effect on Pf. Forskolin stimulated Pf was blocked both by A-779 and by the V2 antagonist. Finally, Ang-(1 7) increased cAMP levels in fresh IMCD cell suspensions whilst the forskolin stimulated cAMP synthesis was decreased by A-779 and the V2 antagonist. These data provide evidence that Ang-(1-7) interacts via its receptor with the AVP V2 system through a mechanism involving adenylate-cyclase activation. PMID- 14534791 TI - Effects of heat stress and mechanical stretch on protein expression in cultured skeletal muscle cells. AB - Effects of heat stress, mechanical stretching or a combination of both on the expression of heat shock proteins (HSPs) and total protein level were studied in a culture system. Rat skeletal muscle cells (L6) were cultured on flexible bottomed culture plates. They were subjected to one of the four following conditions: (1) 97 h incubation at 37 degrees C, (2) 1 h incubation at 41 degrees C followed by 96 h incubation at 37 degrees C, (3) 1 h incubation at 37 degrees C followed by 96 h cyclic stretching (18% of initial length, 2-s stretch and 4-s release) at 37 degrees C or (4) 1 h incubation at 41 degrees C followed by 96 h cyclic stretching at 37 degrees C. The expression of HSP72 and HSP90 and total protein was determined in the crude homogenates, supernatant and pellets. Cellular protein concentrations in the homogenates and pellets were increased by heat stress and/or mechanical stress (stretch). A cumulative effect of the combination of heating and stretch on the protein concentration in the homogenates and in the pellets was noted. The expressions of HSP72 and HSP90 in the pellets were also increased by heat stress and/or stretch. However, HSP90 in the supernatant did not change following heat stress and/or stretch. The regulation of HSP72 and HSP90 expression in skeletal muscle cells may be closely related to total protein, the abundance of which is also stimulated by mechanical and heat stresses. These observations suggest strongly that heating and passive stretch of muscle may be useful as a means of increasing muscle mass, not only in athletes but also in patients during rehabilitation. PMID- 14534792 TI - The cardioprotective effects of Na+/H+ exchange inhibition and mitochondrial KATP channel activation are additive in the isolated rat heart. AB - The mechanisms of recovery of the isolated rat heart were studied after 30 min of global ischemia. Functional recovery was assessed by the percentage recovery of developed pressure after 30 min reperfusion and by the magnitude of the contracture on reperfusion. After a control ischemia, developed pressure recovered to only 12+/-2% of pre-ischemic control and the reperfusion contracture was very large (81+/-6 mmHg). Activation of the mitochondrial KATP channel with 100 microM diazoxide present throughout ischemia and reperfusion improved recovery of developed pressure to 36+/-3% and reduced the reperfusion contracture (53+/-4 mmHg). Inhibition of the sodium/hydrogen exchanger with 10 microM cariporide caused a larger recovery of developed pressure to 72+/-4% and further reduced the reperfusion contracture (11+/-3 mmHg). The combination of both drugs increased recovery of developed pressure to 96+/-4% and the reperfusion contracture remained small (11+/-5 mmHg). The effectiveness of the timing of exposure to these drugs was explored. When both diazoxide and cariporide were applied 2 min before the end of ischaemia and remained present during reperfusion the recovery of developed pressure was 81+/-4% and the reperfusion contracture was small (12+/-3 mmHg); neither was significantly different to the recovery when both drugs were present throughout ischemia and reperfusion. We conclude that mitochondrial damage, blocked by diazoxide, and the coupled exchanger pathway, blocked by cariporide, are two of the principal damage pathways and functional recovery appears to be complete when both are blocked. The combination of these drugs is also highly effective when given 2 min before the end of ischemia. PMID- 14534793 TI - The role of cell type in bone healing mediated by ex vivo gene therapy. AB - BACKGROUND: The ideal cellular vehicle for use in cell-mediated gene therapy to enhance bone healing has not yet been identified. The purpose of this study was to compare the capacity of two types of cells transduced with retro-bone morphogenetic protein 4 (BMP4)-muscle-derived cells (MDCs) and unfractioned bone marrow stromal cells (BMSCs). METHOD: Primary rat MDCs and unfractioned rat BMSCs were transduced with a retrovirus to express BMP4. A 7-mm, critical-sized femur defect was created in adult rats, and 5 x 10(6) transduced cells were implanted into the femoral defect. Bone healing was monitored radiographically and histologically at 4, 8, and 12 weeks post-implantation. RESULTS: All specimens in the MDC-BMP4 group and BMSC-BMP4 group showed a bridging callus at 8 and 12 weeks. At 12 weeks post-implantation the calluses of the MDC-BMP4 femora displayed significantly higher bone photodensity than the BMSC-BMP4 femora (P<0.05). Histomorphometry revealed no difference between the two treatment groups. However, non-union between newly formed and original bone was observed in none of the MDC femora but in six femora from the BMSC-BMP4 group. CONCLUSION: Both MDCs and unfractioned BMSCs can improve healing of a critical-sized bone defect following transduction of the cells with retroBMP4. However, MDCs appear to yield superior results when compared with BMSCs in terms of improved healing of segmental defects. PMID- 14534794 TI - Temporal profile of microvascular disturbances in rat tibial periosteum following closed soft tissue trauma. AB - BACKGROUND AND AIMS: Bone devascularization due to impaired periosteal perfusion following fracture with severe soft tissue trauma has been proposed to precede and underlie perturbed bone healing. The extent and temporal relationship of periosteal microcirculatory deteriorations after severe closed soft tissue injury (CSTI) are not known. We hypothesized that periosteal microcirculation is adversely affected and the manifestation of trauma-initiated microvascular impairment in periosteum is substantially prolonged following CSTI. MATERIAL AND METHODS: Using the controlled-impact injury device, we induced standardized CSTI in the tibial compartment of 35 isoflurane-anesthetized rats. Following the trauma the rats were assigned to five groups, differing in time of analysis (2 h, 24 h, 48 h, 1 and 6 weeks). Non-injured rats served as controls. Before the metaphyseal/diaphyseal periosteum was surgically exposed, intramuscular pressure within tibial compartment was measured. Using intravital fluorescence microscopy (IVM) we studied the microcirculation of the tibial periosteum. We calculated the edema index (EI) by measuring the skeletal muscle wet-to-dry weight ratio (EI = injured limb/contralateral limb). RESULTS: Microvascular deteriorations of periosteal microhemodynamics caused by isolated CSTI were reflected by persistent decrease in nutritive perfusion, markedly prolonged increase in microvascular permeability associated with increasingly sustained leukocyte rolling and adherence throughout the entire study period, mostly pronounced 48 h after the trauma. Peak level in capillary leakage coincided with the maximum leukocyte adherence, tissue pressure, and edema. Microcirculation of tibial periosteum in control rats demonstrated a homogeneous perfusion with no capillary or endothelial dysfunction. CONCLUSION: Isolated CSTI in absence of a fracture exerts long-lasting disturbances in periosteal microcirculation, suggesting a delayed temporal profile in manifestation of CSTI-induced periosteal microvascular dysfunction and inflammation. These observations may have therapeutic implications in terms of preserving periosteal integrity and considering the interaction of skeletal muscle damage and periosteal microvascular injury during management of musculoskeletal trauma. PMID- 14534795 TI - Sequence polymorphism of the mitochondrial DNA control region in the Slovenian population. AB - The forensic application of mitochondrial DNA (mtDNA) typing requires large and regionally well-defined databases. To expand the database for forensic identification purposes in Slovenia, the mtDNA control region sequences of the hypervariable regions HVI and HVII were determined in a population of 129 maternally unrelated Slovenians, using a fluorescent-based capillary electrophoresis sequencing method. A total of 111 different haplotypes resulting from 124 polymorphic positions (80 polymorphic positions in HVI and 44 in HVII) were found. Of these, 101 mtDNA types were unique, 6 haplotypes were shared by 2 individuals, 1 haplotype by 3 individuals, 2 haplotypes by 4 individuals, and the most common haplotype was found in 5 individuals. The most frequent haplotypes in the Slovenian population,263(G), 315.1(C) and 263(G), 309.1(C), 315.1(C) are also the most common in other European populations. The data support the concept that these haplotypes may represent a common European mtDNA sequence types. The sequence poymorphisms were compared to the databases of west Austria and central Italy and the HVI and HVII sequence matching probabilities within and between populations were calculated. It is 1.1-4.5 times more likely to find a sequence match in a random pair of Slovenians than in a random Slovenian-Italian pair and in a random Slovenian-Austrian pair. The length heteroplasmy in the homopolymeric C-stretch regions located at nucleotide positions 16184-16193 in HVI and at positions 303-315 in HVII was observed in 17% and 8% of individuals, respectively. A statistical estimate of the results for this population showed the random match probability and the genetic diversity of 1.16% and 0.996, respectively. PMID- 14534796 TI - Studies on the time frame for ossification of the medial clavicular epiphyseal cartilage in conventional radiography. AB - Radiological assessment of the degree of ossification of the medial clavicular epiphyseal cartilage plays a vital part in forensic age diagnosis of living adolescents and young adults. A total of 873 plain chest radiographs requested by the staff medical officer for members of staff aged 16-30 at the University Hospital Charite were evaluated retrospectively. Of these X-rays 699 permitted an assessment of ossification of at least 1 side of the clavicle. In addition to the customary stages (1: non-ossified epiphysis, 2: discernible ossification centre, 3: partial fusion, 4: total fusion) a stage 5 was also defined, characterised by the disappearance of the epiphyseal scar following total fusion. The earliest age at which stage 3 was detected in either gender was 16 years. Stage 4 was first observed in women at 20 years and in men at 21 years. In both genders, the earliest observation of stage 5 was at 26 years. It was concluded that plain chest radiographs can essentially be used to assess clavicular ossification. In practice, if overlap in posterior-anterior views impedes evaluation, a lateral view should also be taken to facilitate age estimation. In forensic practice the reference values of the present paper should be applied. PMID- 14534797 TI - Diameter of mammary terminal ducts as an additional tool in evaluation of women with polycystic ovarium disease. AB - OBJECTIVE: Aim of this study was to show the change of diameter of mammary ducts in relation to polycystic ovaries disease. DESIGN: Women who took part were divided into two groups- the first with polycystic ovaries disease and another second group with no such disease. METHOD: To both groups a series of laboratory, clinical and ultrasound (u/s) exams were used to diagnose polycystic ovaries disease. Furthermore u/s measurement of mammary ducts was used to define the diameter. RESULTS: In women with polycystic ovaries disease u/s showed an increase in diameter of mammary ducts whereas in the second group we didn't observe such finding. CONCLUSION: Our study showed that there seems to be a strong relation between the increased diameter of the main terminal lactiferous ducts- the polycystic configuration of the ovaries and the hormonal levels. PMID- 14534798 TI - Remodeling capacity and functional outcome of palmarly versus dorsally displaced pediatric radius fractures in the distal one-third. AB - INTRODUCTION: The purpose of this retrospective study was to compare the remodeling capacity and functional outcome of palmarly and dorsally displaced pediatric radius fractures in the distal one-third. MATERIALS AND METHODS: Fifty three children with a residual dorsal angulation of 15 degrees (range 10 degrees 28 degrees, +/-SD 5.32) and 31 children with a residual palmar angulation of 15 degrees (range 10 degrees -30 degrees, +/-SD 4.88) at fracture healing were re examined clinically and radiologically with a median follow-up time of 10 years (range 5-15 years). RESULTS: There was no difference in the remodeling capacity, palmar tilt, radial inclination, and ulnar variance between both groups at follow up. Pain as well as flexion/extension of the wrist and pronation showed no difference in both groups. Palmarly displaced fractures showed a significantly higher restriction of supination ( p=0.01). CONCLUSION: We conclude that remodeling of residual palmar angulation occurs to the same extent as it does in dorsal angulation. Functional outcome differs in forearm supination. PMID- 14534799 TI - Congenital contracture of the quadriceps muscle: a case report with magnetic resonance imaging. AB - INTRODUCTION: Case report of a rare form of congenital contracture of the quadriceps muscle. Congenital contracture of the quadriceps muscle is encountered very rarely in daily orthopaedic practice. A few cases have been reported, but unfortunately these did not detail the MRI findings of congenital contracture. MATERIALS AND METHODS: A 34-year-old woman presented with difficulty in sitting with full flexion of the bilateral knee joints. She had no history of intramuscular injection, and her brother had a similar abnormality. A physical and radiographical review of the case was conducted. RESULTS: A palpable corded induration was detected in the quadriceps muscle which prevented further flexion of the bilateral knee joints. Magnetic resonance imaging of both thighs demonstrated marked atrophy of the rectus femoris muscle and dark signal intensity of the muscle on both T1-weighted and T2-weighted images. It was suggested that the muscles had been replaced by fibrosis. CONCLUSION: This appears to be the first report to include MRI findings of congenital contracture. Clinical awareness of congenital contracture with unique clinical symptoms and radiographic findings may aid the correct diagnosis. PMID- 14534800 TI - Hypomethylation of L1 retrotransposons in colorectal cancer and adjacent normal tissue. AB - BACKGROUND AND AIMS: Malignant cells often exhibit perturbations in the pattern of cytosine methylation. Hypermethylation of CpG islands has been extensively documented, but genome-wide hypomethylation is also a common feature of malignant cells. The bulk of cytosine methylation in the mammalian genome occurs on repetitive elements. This study analysed the methylation status of L1 retrotransposons in colorectal cancer. PATIENTS AND METHODS: Methylation sensitive Southern blotting was used to determine L1 promoter methylation in colon tumours, adjacent normal tissue, and normal colonic mucosa from healthy individuals. RESULTS: Hypomethylation of L1 promoter sequences was detected in all tumours but was also detected in the histologically normal colonic mucosa of 6 of 19 cancer patients, even at a considerable distance from the tumour. L1 hypomethylation was not detected in matched normal peripheral blood, lymph node or smooth muscle tissue from cancer patients or in the colonic mucosa of 14 healthy individuals. We also assayed for the total proportion of methylated CpG in normal bowel specimens from normal and colon cancer patients. Normal mucosa from cancer patients exhibited lower levels of genomic methylation than the mucosa from healthy individuals, and levels were significantly lower in those patients exhibiting L1 promoter hypomethylation. CONCLUSION: These results suggest that genomic hypomethylation is an early event in tumourigenesis. Progressive demethylation of L1 promoter sequences could lead to disturbance of normal gene expression and facilitate the process of neoplastic progression. PMID- 14534801 TI - Defecation mechanisms after proctocolectomy and ileal pouch--anal anastomosis in dogs. AB - BACKGROUND AND AIMS: The role of intrinsic pouch motility after ileal pouch-anal anastomosis (IPAA) during defecation is still speculative. MATERIALS AND METHODS: IPAA was performed in 12 dogs. Defecation mechanisms were evaluated by motility recordings during spontaneous defecation and during expulsion of an endoluminal balloon and by radiography with sequential sector-related gray scale analysis. RESULTS: Spontaneous defecations appeared without significant changes in electrical or mechanical activity of the pouch. Sequential filling of the pouch led to defecation in only seven dogs while the others did not succeed in emptying their pouch even with maximal balloon inflation. Neither strain gauge measurements nor electromyography demonstrated peristaltic contractions of the pouch during defecation while sector-related gray scale analysis revealed strong contractions of the abdominal wall during pouch emptying. CONCLUSION: Pouch emptying is independent of intrinsic pouch motility. The ileoanal pouch acts as a functionally passive reservoir, and its evacuation is initiated by a rise of the intra-abdominal pressure. PMID- 14534803 TI - Preliminary reference levels in interventional cardiology. AB - This article describes the European DIMOND approach to defining reference levels (RLs) for radiation doses delivered to patients during two types of invasive cardiology procedures, namely coronary angiography (CA) and percutaneous transluminal coronary angioplasty (PTCA). Representative centres of six European countries recorded patients' doses in terms of dose-area product (DAP), fluoroscopy time and number of radiographic exposures, using X-ray equipment that has been subject to constancy testing. In addition, a DAP trigger level for cardiac procedures which should alert the operator to possible skin injury, was set to 300 Gyxcm2. The estimation of maximum skin dose was recommended in the event that a DAP trigger level was likely to be exceeded. The proposed RLs for CA and PTCA were for DAP 45 Gyxcm2 and 75 Gyxcm2, for fluoroscopy time 7.5 min and 17 min and for number of frames 1250 and 1300, respectively. The proposed RLs should be considered as a first approach to help in the optimisation of these procedures. More studies are required to establish certain "tolerances" from the proposed levels taking into account the complexity of the procedure and the patient's size. PMID- 14534804 TI - Diffusion-weighted imaging: basic concepts and application in cerebral stroke and head trauma. AB - Diffusion-weighted imaging (DWI) of the brain represents a new imaging technique that extends imaging from depiction of neuroanatomy to the level of function and physiology. DWI measures a fundamentally different physiological parameter compared with conventional MRI. Image contrast is related to differences in the diffusion rate of water molecules rather than to changes in total tissue water. DWI can reveal pathology in cases where conventional MRI remains unremarkable. DWI has proven to be highly sensitive in the early detection of acute cerebral ischemia and seems promising in the evaluation of traumatic brain injury. DWI can differentiate between lesions with decreased and increased diffusion. In addition, full-tensor DWI can evaluate the microscopic architecture of the brain, in particular white matter tracts, by measuring the degree and spatial distribution of anisotropic diffusion within the brain. This article reviews the basic concepts of DWI and its application in cerebral ischemia and traumatic brain injury. PMID- 14534805 TI - Diagnostic management of patients with SAPHO syndrome: use of MR imaging to guide bone biopsy at CT for microbiological and histological work-up. AB - Propionibacterium acnes (P. acnes) is suspected to be involved in the pathophysiology of SAPHO syndrome, since it has been isolated repeatedly through open surgical bone biopsy. This study demonstrates the role of MRI in identifying inflamed bone areas in patients with SAPHO syndrome and the role of CT-guided bone biopsies in obtaining samples from these areas for microbiological and histopathological investigations, thus obviating open surgery. Fourteen consecutive patients with SAPHO syndrome were investigated by MRI to identify acute inflammatory changes in hyperostotic periarticular bone. The CT-guided biopsies for microbiological investigations were taken from the areas identified. Patients positive for P. acnes were started on long-term antibiotic therapy according to antibiotic susceptibility. On MRI the inflammatory changes appeared as hyperintense areas on fat-saturated T2 fast-spin-echo (FSE) images and showed signal increase on fat-saturated T1 SE images after Gd-DTPA. With MR localization CT-guided bone biopsies yielded P. acnes in 8 patients. No bacteria could be isolated from the remaining 6 patients. Acute inflammatory bone changes in SAPHO syndrome are well localized by MRI. With MR localization, CT-guided bone biopsies offer a minimally invasive alternative to open surgery in the detection of. P. acnes leading to the institution of a specific antibiotic therapy. PMID- 14534806 TI - Influence of the radiographer on the pain felt during mammography. AB - Mammography is the only useful examination in screening for breast cancer. Mortality from breast cancer can be reduced if women go regularly for a screening mammography. Moreover, it is still the key examination in diagnosis of breast diseases and in the follow-up of patients treated for breast cancer. Pain with mammography can deter women from going for regular screening or follow-up; therefore, it is important to reduce pain experience or discomfort from mammography. In this study we evaluate the impact of the "radiographer" on the pain risk during mammography by analysing questionnaires filled in by women and radiographers. Study results reveal that the opinion of the radiographer, the information and communication during the examination and the number of years of experience are important factors in pain and discomfort experience. The attitude of the radiographer plays an important role in the pain experience. PMID- 14534807 TI - Visually lossless threshold determination for microcalcification detection in wavelet compressed mammograms. AB - The aim of this study was to determine the visually lossless threshold of a wavelet-based compression algorithm in case of microcalcification cluster detection in mammography. The threshold was determined by means of observer performance using a set of digitized mammograms. In addition, the transfer characteristics of the compression algorithm were assessed by means of image quality parameters using computer-generated test images. The observer performance study was based on rating performed by four independent radiologists, who reviewed 68 mammograms, from the Digital Database for Screening Mammography (DDSM), at six different compression ratios. Receiver operating characteristics (ROC) analysis was performed on observers' responses and the area under ROC curve (A(z)) was calculated at each compression ratio for each observer. The parameters used for assessment of transfer characteristics of the compression algorithm were input/output response, noise, high-contrast response, and low-contrast-detail response. The computer-generated test image, used for this assessment, mimicked mammographic image characteristics (pixel size, pixel depth, and noise) as well as microcalcification characteristics (size and contrast). The ROC analysis for microcalcification cluster detection indicated a threshold at compression ratio 40:1, as Student's t-test shows statistically significant differences in A(z) values (p<0.05) for compression ratios 70:1 and 100:1. Observers' grading of mammogram quality lowers this threshold at 25:1. Low-contrast-detail detectability in the transfer characteristics study indicate a threshold of 35:1, whereas non-perceptibility of image-quality-parameters degradation lowers this threshold to 30:1. The ROC and transfer characteristics analysis provided comparable thresholds, indicating the potential use of the latter in limiting the target range of compression ratios for subsequent observer studies. PMID- 14534810 TI - Antibiotic prevention of pneumococcal infections in asplenic hosts: admission of insufficiency. AB - Asplenic and hyposplenic patients have an increased risk for overwhelming pneumococcal infections, even several decades after splenectomy. Pneumococcal vaccination and daily oral administration of penicillin V are recommended to prevent such infections, 2-5 years after splenectomy, and for at least 5 years in children affected with sickle cell disease. In order to assess whether the infectious risk is actually known and prevented, we interviewed physicians (belonging to a general practitioner and pediatrician network) who followed patients having undergone a splenectomy and/or children with sickle cell disease under 5 years of age. We received replies from 104 physicians monitoring 152 patients replied. Potential infection risk was not known for 28% of the asplenic patients and 40% of the children with sickle cell disease. Only 75% of the asplenic patients and 36% of the children with sickle cell disease had been vaccinated against pneumococcus. Of the patients who had undergone splenectomy, 27% had been treated with an antibiotic after surgery and 60% had discontinued it, the vast majority of them during the same year. Of the children with sickle cell disease, 48% were not receiving an antibiotic. This study demonstrates that risk of infections in asplenic patients is widely misunderstood, indicating the urgent need to improve their management. PMID- 14534808 TI - Diagnostic performance of liquid crystal and cathode-ray-tube monitors in brain computed tomography. AB - The aim of this study was to evaluate feasibility of reporting brain CT examinations on liquid crystal display (LCD) flat-screen monitors vs state-of-the art cathode-ray-tube (CRT) monitors. Ninety-five brain CT examinations of 95 patients were displayed on Picture archiving and communications system (PACS) workstations equipped either with a dedicated medical imaging LCD colour monitor or on a high-resolution CRT which is used for routine reporting of CT, MRI and digital radiography images in our institution. Fifty cases were negative and 45 cases were positive for early brain infarction (EBI), the latter being defined by a combination of one or more signs: dense artery; hypodensity of brain parenchyma; and local brain swelling verified by control scans. Ten radiologists had to rate presence or absence of EBI on a five-point scale. Ratings were evaluated by CORROC2 ROC software and areas under the ROC curve (A(z)) were computed. Significance of differences between the two viewing conditions were evaluated with Wilcoxon test. Mean A(z) of the ten observers was 0.7901 with LCD vs 0.7695 with CRT which did not show statistical significance (p=0.2030). In the setting investigated, reporting of CT studies from high-performance LCD monitors seems feasible without significant detriment to diagnostic performance. PMID- 14534817 TI - Definition and classification of chronic pancreatitis. AB - Chronic pancreatitis has been defined by features that are clinical and etiologic in origin. From such definitions a number of classifications have been developed, starting with the Marseille Classification in 1963. Defining features of chronic pancreatitis must consider the etiology of the disease, which then presents characteristic clinical evidence through symptomatic sequelae of organ pathology, including complications of the disease. Advancing technology has improved the imaging capabilities available to document derangements of the pancreas. The main purpose of international classifications of disease states is to provide a common language for purposes of clinical, research, and health outcomes studies. Modification of the 10th World Health Organization I.C.D. classification may provide this tool. PMID- 14534819 TI - Lateral pancreaticojejunostomy for chronic pancreatitis. AB - Chronic pancreatitis is a progressive fibrosis of the pancreas that leads to loss of endocrine and exocrine function. The most common symptom in this disease is intractable pain. The etiology of pain in chronic pancreatitis is not clearly understood. However, many of these patients have dilated ducts consisting of saccular dilations and intervening constructions referred to as the "chain of lakes" phenomenon. These patients can be diagnosed with either endoscopic retrograde cholangiopancreatography (ERCP) or computed tomography (CT). These patients are best treated by the Partington Rochelle modification of the Puestow Procedure otherwise known as lateral pancreaticojejunostomy. Overall pain relief in published studies occurs in 50-90% of patients. Another proposed advantage of the lateral pancreaticojejunostomy is preservation of endocrine and exocrine pancreatic function as long as the pancreas is not further damaged by alcohol. PMID- 14534818 TI - Treatment of pancreatic exocrine deficiency. AB - In treating pancreatic exocrine insufficiency the diagnosis of a cause, wherever possible, is a first step. If there is an obstructing lesion, then its relief may be curative. Adequate management depends on dietary manipulation and enzyme replacement. A reduction in fat intake, with accentuation of protein and carbohydrate consumption, is recommended. For enzyme replacement, enteric-coated microspheres are required, taken with adequate acid-suppression with a proton pump inhibitor. The enzyme supplements, using standard preparations of pancreatin, usually of porcine source, may be as much as 4 to 8 tablets per meal and should be taken with food. Endocrine insufficiency usually requires insulin. PMID- 14534820 TI - Pancreatoduodenectomy in the treatment of chronic pancreatitis. AB - The value of surgical resection for patients with chronic pancreatitis has been debated on account of persistent symptoms and the morbidity of the operative procedure, both immediate and delayed. This paper explores the consequence of pancreatoduodenectomy in 175 patients with chronic pancreatitis who were operated on between 1976 and 1997. All patients were carefully selected after varying periods of conservative management. The operation was a classical Kausch-Whipple resection in 98 patients and a pylorus-preserving procedure in 67. There were four postoperative deaths (days 7, 10, 35, and 70), and only two reoperations were performed. The median number of postoperative events was one, with delayed gastric emptying being the most common (31 patients). The median length of in hospital stay was 20 days (range: 8-215 days), but no patient was discharged until medical and social disabilities were resolved. There were seven late deaths, most of them linked to cigarette smoking and alcohol consumption; 75% of patients had a good clinical outcome, but 18 patients required further pancreatic surgery at a mean of 12 months, either a pancreatojejunostomy or a completion pancreatectomy. Diabetes occurred in 40% of patients by 5 years, and most, at some stage of their postoperative period, required pancreatic enzyme supplementation. It is suggested that resection of the pancreas provides a reasonable life-style in 75% of patients, but the outcome depends in large part on the predisposing disease. PMID- 14534822 TI - Subtotal pancreatectomy for chronic pancreatitis. AB - Chronic pancreatitis results when pancreatic structure or function is irreversibly damaged by repeated or ongoing inflammation, regardless of the underlying etiology. Most patients present with medically intractable pain and radiological evidence of diffuse gland involvement. Surgical therapy is directed mainly toward palliation of symptoms, and cure is unusual except when the inflammatory process is limited to a specific segment of the pancreas. Surgical strategy should be individualized on the basis of alterations in pancreatic morphology and duct anatomy. In properly selected patients, duct drainage procedures effectively relieve pain and preserve pancreatic function with low perioperative morbidity and mortality. Extensive distal pancreatectomy is effective in relieving pain, but it can be technically challenging and in general should be limited to patients with small-duct disease because of severe metabolic consequences. Intraportal islet cell autotransplantation or segmental pancreatic autotransplantation may ameliorate the long-term effects of insulin-dependent diabetes, but it will have limited applicability until methods for optimizing and purifying islets are developed and the optimal route and site of islet cell implantation have been identified. PMID- 14534821 TI - Comparison of local resection of the head of the pancreas combined with longitudinal pancreaticojejunostomy (frey procedure) and duodenum-preserving resection of the pancreatic head (beger procedure). AB - The etiology of pain in chronic pancreatitis may be ductal hypertension, increased parenchymal pressure, or neural damage. It is difficult to assess the severity of pain in this patient population, a problem made more challenging by the frequency of narcotic dependency. Therapeutic interventions developed to relieve the pain of chronic pancreatitis include denervation of the pancreas, decompression of the main duct of the pancreas, resection of part or all of the diseased pancreas, and reduction of pancreatic secretion. Operative intervention for patients with chronic pain is indicated when severe pain, complications of pain, or potential malignancy are present. The operations that consistently provide long-lasting pain relief all have in common resection of all or a portion of the head of the pancreas. Adverse effects on exocrine and endocrine function, nutrition, and quality of life are related to the amount of pancreas resected. The ideal procedure should be easy to perform, have a low morbidity and mortality rate, provide long-lasting pain relief, and not augment endocrine and exocrine insufficiency. No single operation fulfills this ideal. The local resection of the head of the pancreas combined with longitudinal pancreaticojejunostomy (LR LPJ) proposed by Frey and the duodenum-preserving resection of the head of the pancreas (DPHR) proposed by Beger are discussed. The conceptualization, development, and technique of LR-LPJ are discussed, and comparisons of patient outcomes are made with the outcomes of other procedures for chronic pancreatitis. PMID- 14534823 TI - Nerve blocks and neuroablative surgery for chronic pancreatitis. AB - Surgical decompressive procedures for "large-duct" chronic pancreatitis have been notably successful in relieving pain. However, management of patients with intractable pain from "small-duct" chronic pancreatitis has been difficult, often resulting in narcotic addiction and/or malnutrition from major pancreatic resection. In view of the disappointing results from extensive pancreatic resections in these cases, denervation of pancreatic sympathetic pain afferents has been suggested as an alternative. Although denervation procedures have been attempted at multiple anatomical levels, results have unfortunately been mixed. The observed variability in results has been attributed to poor patient selection, incomplete understanding of neurophysiology of pancreatic pain, and perhaps inadequate knowledge of pancreatic neuroanatomy. At present, the preferred form of neural ablation is splanchnicectomy. However, a consistent and reliable method for identifying candidates for splanchnicectomy is critical, as it is clinically difficult to distinguish true pancreatic pain from other nociceptive conditions masquerading as pancreatic pain. Differential epidural anesthesia (DEA) is a promising, safe test for initial evaluation and patient selection, although it is not as precise as sometimes claimed. Patients responding to sympathetic block during DEA seem to be the best candidates for operative sympathetic ablation. At the moment, the optimal surgical approach to splanchnic ablation, which offers the least morbid technique, most favorable results, and an attractive risk-benefit ratio, is bilateral thoracoscopic splanchnicectomy. More experience and longer follow-up will be necessary to evaluate this approach. PMID- 14534825 TI - Modified alar swing procedure in saddle nose correction. AB - Reconstruction of the saddle nose may involve the use of different augmentation materials, from autogenous bone and cartilage to alloplastic materials. The most important problems when considering the choice of reconstructive technique, besides underlying pathology and expected result, include: long-term stability, donor morbidity, tendency of the implant to infection, extrusion, and resorption. The use of the lateral crura of the lower lateral cartilages as dorsal onlay was reserved for the corrections of minor supratip depressions (flying wing and alar swing procedure). The authors suggest the use of pedicled flaps of cephalic portions of lateral crura as dorsal septal strut, which may increase the profile line more than dorsal onlay. Reconstruction is performed using open rhinoplasty approach. Pedicled flaps of the cephalic portions of lateral crura are transfixed in the sagittal plane and, following separation of upper lateral cartilages and medial crura, placed on the dorsum of nasal septum. Upper laterals are sutured to newly formed cartilaginous dorsum, or a new bridge is created using conchal cartilage. Columellar strut may be formed of the septal cartilage. Authors have performed such corrections in 15 patients with good long-term functional and aesthetic results. PMID- 14534824 TI - Management of biliary and duodenal complications of chronic pancreatitis. AB - Biliary stricture and duodenal obstruction have been increasingly recognized as complications of chronic pancreatitis. The anatomical relationship of the distal common bile duct and the duodenum with the head of the pancreas is the main factor for their involvement in chronic pancreatitis. In hospitalized patients with pancreatitis, the incidence of biliary stricture and duodenal obstruction is reported to be about 6% and 1.2%, respectively. For patients requiring an operation for chronic pancreatitis the incidence increases to 35% for biliary stricture and 12% for duodenal obstruction. Fibrosis around the distal common bile duct can cause stenosis with obstruction of bile flow. Clinically, the presentation of these patients ranges from being asymptomatic with elevated alkaline phosphatase or bilirubin, or both, to being septic with cholangitis. Jaundice, cholangitis, hyperbilirubinemia, and persistent elevation of serum alkaline phosphatase occur more frequently in patients with pancreatitis with a biliary stricture. A twofold elevation of alkaline phosphatase is a marker of possible common duct stenosis in patients with chronic pancreatitis. The incidence of both biliary cirrhosis and cholangitis in these patients is about 10%. ERCP reveals a characteristic long, smoothly tapered stricture of the intrapancreatic common bile duct. In duodenal obstruction, the factors that convert self-limiting edema to chronic fibrosis and stricture formation are unknown, but ischemia superimposed on inflammation may be the major cause. These patients present with a prolonged history of nausea and vomiting. Barium studies typically show a long constricting lesion of the duodenum, and endoscopy reveals reactive inflammatory changes in a narrowed duodenum. Operation is indicated in patients with common bile duct strictures secondary to chronic pancreatitis when there is evidence of cholangitis, biliary cirrhosis, common duct stones, progression of stricture, elevation of alkaline phophatase and/or bilirubin for over a month, and an inability to rule out cancer. The operation of choice is either choledochoduodenostomy or choledochojejunostomy. A cholecystoenterostomy is less favored because of its higher failure rate (23%). Endoscopic stenting plays a role in patients who are unfit for surgery, but it is not recommended as definitive therapy. For duodenal obstruction, failure to resolve the obstruction with 1-2 weeks of conservative therapy is an indication for bypass. The operation of choice is a gastrojejunostomy. Not uncommonly, combined obstruction of the pancreatic duct, common bile duct, and duodenum will develop. Combined drainage procedures or resection are used to manage these problems. PMID- 14534826 TI - Temporal lift by galeapexy: a review of 270 cases. AB - The purpose of this technique is to correct the ptotic tail of the brow and crow's feet as well as to redrape the premalar skin to allow us to make a greater skin resection in the lower lid. The purpose of this presentation is to expose the advantages, the inconveniences, and the complications of this technique. We reviewed the charts of 270 patients who were operated on between June 1996 and January 2002. The average age of the patients was 45 years old. Temporal lifts were performed during a frontotemporal lift in 92 cases, and during a three-stage lift (frontal, facial, and cervical) in 170 cases. In eight cases, an isolated temporal lift was performed. An upper blepharoplasty was performed in 252 cases and transconjunctival fat pad excision was performed in 188 cases. Finally, a lower lid skin resection was done following a galeapexy in 241 cases. The surgical approach consists of a temporal incision, followed by a subgaleal dissection, and then a subcutaneous dissection that allows the cephalic galea, previously incised, to be anchored to the temporal aponeurosis. This allows good cutaneous redraping that raises the brow tail and a detachment of the orbicularis fibers. The suturing of the scalp is done without tension and without eliminating hair. The results of the technique with a follow-up period of more than five years for the earlier cases and six months for the last cases, were evaluated with three criteria: (1) the distance between the brow and the eyelashes, (2) the reduction of crow's feet, (3) the reduction of wrinkles at the level of the lower lid and the malar region. Overall satisfaction was also rated. Of 270 patients, 225 had ratings of very good to good, 13 had average results, and 13 had unsatisfactory results. As far as complications, five had unilateral hematoma requiring an evacuation, four patients had temporary alopecia, and eight patients had temporary frontal muscle weakness that resolved within one to three months. This technique, performed under local anesthesia, is very efficient, reproducible, and simple. The incidence of complications can be reduced when one is meticulous during dissection. PMID- 14534827 TI - Mammaplasty: a new approach. AB - In this paper, the authors demonstrate a single stitch that brings together and encircles the mammary base at the aponeurosis of the musculus pectoralis major, closing the mammary base while elevating the mammary cone and achieving better projection of the superior poles. PMID- 14534829 TI - Primary Stanmore total hip arthroplasty with increased cup loosening in rheumatoid patients. AB - We studied all Stanmore total hip arthroplasties (THA, n=325) implanted at our center between 1980 and 1990. Seven patients (seven hips), followed for less than 12 months, were excluded. Five hips were lost at follow-up. The mean follow-up was 117 (12-252) months. A total of 18 hips were revised, with median time between operation and revision 163 months. With revision as endpoint, a survival rate analysis was calculated. The mean survival for all hips was 82% at 18 years (95% confidence interval 64-101%). For hips with rheumatoid arthritis, the survival rate was 58%, whereas in osteoarthritic hips the rate was 95% after 18 years. A high proportion of acetabular loosening was seen in rheumatoid hips. PMID- 14534830 TI - The femoral cement mantle in three total hip replacements. AB - We measured the size and volume of the femoral broaches and stems supplied with three commercial total hip replacement (THR) systems (JRI Furlong, Biomet Stanmore, and Howmedica Exeter). Using an in vitro method, we created cement mantles that could be directly measured. The broaches supplied with the Biomet Stanmore and the Howmedica Exeter systems allowed the creation of significantly thicker cement mantles than the JRI broaches and stems (Stanmore mean thickness = 1.7 mm, defects at 3% of measurement sites; Exeter mean thickness = 2.0 mm, no defects; JRI mantles mean thickness = 0.9 mm, cement defects at 29% of measurement sites). We conclude that, in vivo, the broaches supplied with the Stanmore and Exeter systems are significantly larger than the corresponding stems, and will excavate a cavity large enough to accommodate the appropriate femoral stem surrounded by an intact cement mantle. PMID- 14534831 TI - Glove punctures in orthopaedic surgery. AB - From February 2001 to May 2001, 792 latex gloves used in 100 operations from three orthopaedic sub-specialties (paediatrics, hand and spine) were tested for puncture by a water infusion test. Sixty-nine gloves from 45 operations were punctured, giving an overall glove perforation rate of 8.7% (69/792) and an operative perforation rate of 45% (45/100). The hand operations had the lowest operative perforation rate (19.4%) while the spine operations had the highest (63.6%). The glove perforation rate increased in bony procedures (60% versus 22.5%), in procedures with major instrumentation (66% versus 18%) and in more lengthy procedures. The thumbs and the left index finger had more punctures than other parts. In addition, the glove perforation rate for single gloving was 9.6% (53/552) while that for the inner glove of double gloving was 0.8% (1/120). Based on these findings, we would like to recommend double gloving and regular glove changing in these high-risk surgeries. PMID- 14534832 TI - Characterisation of [123I]iomazenil distribution in a rat model of focal cerebral ischaemia in relation to histopathological findings. AB - Iodine-123 labelled iomazenil ([(123)I]IMZ) has been reported to be a useful marker of neuronal viability. The brain distribution of [(123)I]IMZ, however, has not been correlated with the pathophysiological response in detail after an ischaemic insult. To characterise [(123)I]IMZ as a marker of neuronal viability, we compared its brain distribution with cyclooxygenase-2 (COX-2) expression, DNA fragmentation and cellular integrity. [(123)I]IMZ and [(125)I]IMP were injected into rats with focal cerebral ischaemia for the purpose of dual-tracer autoradiography. COX-2 and microtubule-associated protein-2 (MAP-2, a marker of cellular integrity) were immunostained. In situ DNA polymerase-I-dependent dUTP incorporation into damaged DNA was used as an indicator of DNA fragmentation. Lesion to normal ratios (LNRs) for [(123)I]IMP and [(125)I]IMZ were calculated. [(123)I]IMZ accumulation was preserved in several regions with impaired [(123)I]IMP accumulation. COX-2 expression was occasionally observed, whereas neither DNA fragmentation nor MAP-2 denaturation was detected in these regions. DNA fragmentation and impaired MAP-2 immunostaining were observed only in the regions with reduced LNRs for both tracers. The LNR for [(123)I]IMZ was significantly lower in regions with impaired MAP-2 immunostaining (0.120+/-0.152, P<0.0001), in regions positive for dUTP incorporation (0.488+/-0.166, P<0.0001) and in regions positive for COX-2 expression (0.626+/-0.186, P<0.001) than in histologically normal regions (0.784+/-0.213). Thus, neuronal DNA is still intact and cellular integrity is maintained in the ischaemic regions with preserved [(123)I]IMZ accumulation. The impairment of [(123)I]IMZ accumulation precedes DNA fragmentation and denaturation of cellular integrity. These results provide the molecular basis of [(123)I]IMZ distribution. PMID- 14534834 TI - Respiratory motion artefacts on PET/CT. PMID- 14534833 TI - Radiation exposure during transmission measurements: comparison between CT- and germanium-based techniques with a current PET scanner. AB - In positron emission tomographic (PET) scanning, transmission measurements for attenuation correction are commonly performed by using external germanium-68 rod sources. Recently, combined PET and computed tomographic (CT) scanners have been developed in which the CT data can be used for both anatomical-metabolic image formation and attenuation correction of the PET data. The purpose of this study was to evaluate the difference between germanium- and CT-based transmission scanning in terms of their radiation doses by using the same measurement technique and to compare the doses that patients receive during brain, cardiac and whole-body scans. Measurement of absorbed doses to organs was conducted by using a Rando Alderson phantom with thermoluminescent dosimeters. Effective doses were calculated according to the guidelines in the International Commission on Radiation Protection Publication Number 60. Compared with radionuclide doses used in routine 2-[fluorine-18]-fluoro-2-deoxy-D-glucose PET imaging, doses absorbed during germanium-based transmission scans were almost negligible. On the other hand, absorbed doses from CT-based transmission scans were significantly higher, particularly with a whole-body scanning protocol. Effective doses were 8.81 mSv in the high-speed mode and 18.97 mSv in the high-quality mode for whole-body CT based transmission scans. These measurements revealed that the doses received by a patient during CT-based transmission scanning are more than those received in a typical PET examination. Therefore, the radiation doses represent a limitation to the generalised use of CT-based transmission measurements with current PET/CT scanner systems. PMID- 14534838 TI - The effect of epidural free fat graft on the outcome of lumbar disc surgery. AB - Numerous materials have been used to prevent epidural scar tissue after lumbar disc surgery. Free fat grafts are common both experimentally and clinically, but there is some doubt about their protection against fibrosis, and some complications have been reported. In this prospective study, the usefulness of free fat grafts during lumbar disc surgery was evaluated. Ninety-nine patients who had undergone operation due to lumbar disc herniation were divided in two groups: those with implantation of free fat grafts (group A) and those without (group B). Outcome was evaluated at a mean of 2.6 years postoperatively according to the following criteria: visual analog scale for back and leg pain, Hannover Questionnaire on activities of daily living, reflex findings, sensory and motor deficits, consumption of analgesics, walking distance, straight leg raising test, and clinical examination. The outcome variables showed no significant differences between the two groups ( P>0.05). This study suggests that the use of free fat grafts during lumbar disc surgery was clinically ineffective. PMID- 14534835 TI - The caspase pathway in noise-induced apoptosis of the chinchilla cochlea. AB - We previously reported that intense noise exposure causes outer hair cell (OHC) death primarily through apoptosis. Here we investigated the intracellular signal pathways associated with apoptotic OHC death. Chinchillas were exposed to a 4 kHz narrowband noise at 110 dB SPL for 1 h. After the noise exposure, the cochleas were examined for the activity of each of three caspases, including caspase-3, 8, or -9 with carboxyfluorescein-labeled fluoromethyl ketone (FMK)-peptide inhibitors. The cochleas were further examined for cytochrome c release from mitochondria by immunohistology and for DNA degradation by the TUNEL method. The results showed that the noise exposure triggered activation of caspase-3, an important mediator of apoptosis. The noise exposure also caused the activation of caspase-8 and caspase-9, each of which is associated with a distinct signaling pathway that leads to activation of caspase-3. Caspase activation occurred only in the apoptotic OHCs and not in the necrotic OHCs. These results indicate that multiple signaling pathways leading to caspase-3 activation take place simultaneously in the apoptotic OHCs. In addition to caspase activation, noise exposure caused the release of cytochrome c from mitochondria, resulting in a punctate fluorescence in the cytosol. In contrast to activation of caspases, the release of cytochrome c took place in both apoptotic and necrotic OHCs. Moreover, the release of cytochrome c in a subpopulation of OHCs took place early in the cell death process, prior to any outward signs of necrosis or apoptosis. These data suggest that in this subpopulation there exists a common step that is shared by cell death pathways before entering either necrosis or apoptosis. Lastly, use of the TUNEL assay in combination with PI labeling provides a more accurate discrimination between apoptosis and necrosis. PMID- 14534839 TI - Sulfite and base for the treatment of familial amyloidotic polyneuropathy: two additive approaches to stabilize the conformation of human amyloidogenic transthyretin. AB - Recently, we presented evidence that sulfite protects transthyretin (TTR) from normal human individuals and heterozygotes with amyloidogenic TTR mutations against the decay of tetramers into monomers. In this paper we demonstrate a stabilizing effect of sulfite on TTR tetramers from a familial amyloidotic polyneuropathy (FAP) patient homozygous for the most-common amyloidogenic TTR-V30 M mutation. We compare the conformational stability of partially sulfonated TTR from a heterozygote for normal TTR and amyloidogenic TTR-V30 M with the stability of untreated TTR from a compound heterozygote for amyloidogenic TTR-V30 M and TTR T119 M known to have only minor or no problems of FAP. Using a combination of polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate (SDS) gradient PAGE we demonstrate that TTR dimers containing amyloidogenic TTR mutations decay into monomers at pH<7.4. Increasing the pH by some 0.2 units within physiological ranges, i.e., pH 7.0-7.4, and sulfonation of TTR were observed to have additive inhibitory effects on the transition of dimers into monomers. We conclude that mild acidifying episodes in the interstitial volume of tissues at risk for amyloidosis could contribute to the development of FAP. Early and permanent efforts to counteract acidosis by treatment with base could possibly help to delay the onset of the disease. The intake of sulfite could support these efforts. PMID- 14534840 TI - Absence of pathogenic mutations in presenilin homologue 2 in a conclusively 17 linked tau-negative dementia family. PMID- 14534841 TI - Treatment of six hepatic artery aneurysms. AB - Hepatic artery aneurysms are rare lesions but of significant clinical importance because rupture is associated with elevated mortality. Although diagnosis using CT scanning and, more importantly, angiography has been well defined, the therapeutic choices are less clear. We retrospectively selected patients from 1985 to 2000 who were treated with either traditional surgical or percutaneous techniques. In total we treated six patients, four males and two females. This represents 17.6% of 34 patients treated by us for splanchnic artery aneurysms. The treatment was surgical in two cases (33.3%) and percutaneous in four cases (66.6%). All lesions were successfully treated with exclusion/ablation of the aneurysm. Mortality was nil; in one of the surgical cases we reported a transient hepatic failure and in the endovascular group, one right pleural effusion, one small splenic infarction, and one pseudoaneurysm of the gastroduodenal artery. The first therapeutic strategy to be taken into consideration is always the percutaneous approach. However, surgery still has a role in those cases where the lesion cannot be repaired percutaneously. Based on our own experience (good results, no mortality, and few complications) and in accordance with the literature, it is clear that an aggressive approach is warranted in those patients. PMID- 14534842 TI - Evolving complexity of open aortofemoral reconstruction done for occlusive disease in the endovascular era. AB - Available endovascular and less invasive surgical interventions have diminished the need for aortofemoral bypass (AFB) construction for chronic inflow occlusive disease but have potentially increased its complexity. We reviewed our results with AFB done in 107 consecutive patients between 1997 and June 2002 (83 men, 24 women, mean age 62 +/- 7 years) with chronic limb ischemia due to aortoiliofemoral occlusive disease. Perioperative factors and surgical outcomes (<30 days) were evaluated and compared between patients requiring complex (redo AFB, need for visceral aortic clamp for juxtarenal occlusion, adjunctive visceral revascularization, or simultaneous inflow/outflow bypass) and conventional reconstructions by contingency table analysis. AFB was done for limb threat in 65 patients (61%) and 44 patients (41%) had failed previous inflow procedures (22 endovascular, 43 open; 1.5/patient). Operative complexity (36 patients, 34%) was evidenced by the need for redo AFB in 8 patients, suprarenal (13) or supramesenteric/celiac (6) aortic clamp and pararenal endarterectomy in 19 cases, adjunctive renal (10) or mesenteric (2) revascularization, or simultaneous construction of AFB and femoropopliteal/tibial bypasses in 9 patients. Overall AFB operative mortality and major complication rates were 3.7% ( n = 4) and 34% ( n = 36), respectively. Mortality ( p = 0.32) and nonvisceral related complications ( p = 0.3) were not statistically more frequent after complex AFB (5.6%, 31%) than after conventional reconstructions (2.8%, 21%). However, renal, mesenteric, or spinal cord (visceral) ischemic complications or death (10.3%) were greater after complex reconstructions (19.4%) than after conventional AFB (5.6%) ( p = 0.03). Pre-existing renal insufficiency (Cr >/=1.5, n = 9) was not predictive of postoperative renal failure (>2x preop Cr, n = 7) in this series ( p = 0.4). Our recent experience with AFB suggests its increasing use as a tertiary modality after failed endovascular or less invasive open reconstructions. Despite the added operative complexity associated with manipulation of the visceral aorta and its branches and the need for extended infrainguinal revascularization, satisfactory clinical outcomes can be achieved. PMID- 14534843 TI - Outcomes following distal bypass graft occlusion in diabetics. AB - The objectives of this study were to define clinical outcomes following distal bypass graft occlusion in diabetic patients and identify factors predictive of limb loss. A retrospective review was conducted of all distal graft occlusions over a 5-year period in diabetic patients. Popliteal grafts, perioperative (30 days) failures, and redo distal bypasses were excluded. Eighty grafts were studied (43 cases done for ulcers, 31 for gangrene, and 6 for rest pain). Time to occlusion averaged 13 +/- 17 months post-bypass (median 6 months). Interventions following graft occlusion (lysis, thrombectomy, revision, or new bypass) were carried out in only 26 cases, leading to limb salvage in 11 cases (42%). This compares to a limb salvage rate of 54% in the 54 cases where no intervention was done ( p = NS). The overall amputation rate was 50% (14 AKAs and 26 BKAs). The interval between graft occlusion and amputation averaged 2 +/- 2 months. Limbs saved were followed an average of 14 +/- 11 months after graft occlusion. The presence of a foot wound (new or old) was associated with a higher likelihood of amputation (67% vs. 32% for cases with no foot wounds at the time of bypass failure; p < 0.005). In cases requiring an amputation, the toebrachial index averaged 0 +/- 0 post-bypass failure vs. 0.2 +/- 0.2 in cases where limbs were saved ( p < 0.05). Patients with congestive heart failure or who were nonambulatory at the time of graft occlusion were more likely to end up with an amputation ( p < 0.05). Limb loss was also more likely when gangrene had been the initial operative indication. No other factors were predictive of limb loss, including vein configuration, inflow or outflow level, prior bypass revision, known graft stenosis, time to occlusion, age, gender, other medical conditions, pre- or post-bypass ABI or TBI, graft velocities, or use of anticoagulants. Limb loss following primary distal bypass failure in diabetics is high. Repeat interventions have limited success. The findings justify aggressive programs of graft maintenance, and vigorous attempts at early closure of foot wounds. PMID- 14534844 TI - A prospective randomized evaluation of negative-pressure wound dressings for diabetic foot wounds. AB - Optimal treatment for large diabetic foot wounds is ill defined. The purpose of this study was to compare the rate of wound healing with the Vacuum Assisted Closure device trade mark (VAC) to conventional moist dressings in the treatment of large diabetic foot wounds. Diabetics with significant soft tissue defects of the foot were considered for enrollment. Patients were randomized to receive either moist gauze dressings or VAC treatments for 2 weeks, after which they were treated with the alternative dressing for an additional 2 weeks. Wounds were photographed weekly and wound dimensions calculated in a blinded fashion with spatial analysis software. Percent change in wound dimensions were calculated and compared for each weekly assessment and over 2 weeks of therapy with each dressing type. Ten patients were enrolled in the trial, but two were lost to follow-up and two were withdrawn. Complete data were available for analysis on seven wounds in six patients. Average length, width, and depth of the wounds at initiation of the trial was 7.7, 3.5, and 3.1 cm, respectively. Only the wound depth was significantly decreased over the weeks of the trial to 1.2 cm ( p < 0.05). VAC dressings decreased the wound volume and depth significantly more than moist gauze dressings (59% vs. 0% and 49% vs. 8%, respectively). VAC dressings were associated with a decrease in all wound dimensions while wound length and width increased with moist dressings. In summary, over the first several weeks of therapy, VAC dressings decreased wound depth and volume more effectively than moist gauze dressings. Negative-pressure wound treatment may accelerate closure of large foot wounds in the diabetic patient. PMID- 14534845 TI - Carotid artery stenting in a community setting: experience outside of a clinical trial. AB - Carotid artery angioplasty and stenting (CAS) currently represents a less invasive percutaneous alternative to conventional endarterectomy for the treatment of carotid occlusive disease. We report here the results and complication rates of CAS performed by a team of interventionalists at a non clinical trial center utilizing a standardized CAS protocol. CAS was attempted in 51 arteries in 48 patients (mean age 71 +/- 9 years, range 53-90). Fifteen (29%) of 48 patients were symptomatic. Indications for CAS were previous ipsilateral endarterectomy (15/51, 29%), previous neck radiation therapy (1/51, 2%), or significant coronary artery disease (30/51, 59%). SMART((R)) stents were deployed via percutaneous femoral artery access, with anticoagulation (heparin, abciximab, aspirin, clopidogrel) and temporary transvenous cardiac pacemakers employed in all patients. Neuroprotection was not used. Neurological examination and duplex scans were performed in follow-up. CAS was successfully performed in 96% of cases (49 lesions/46 patients) with angiographic stenoses of 88 +/- 8%. Neurological complications included one (2%) minor stroke that occurred 12 hr after CAS. There were no periprocedural mortalities. Clinically significant bradycardia or asystole occurred in 11/49 (22%) procedures, necessitating short-term ventricular pacing. All stented vessels remained patent during 12.2 +/- 10.1 (range 1-37) months follow-up period. One asymptomatic restenosis (>70%) occurred at 3 months, which was successfully reangioplastied; we thus had 1-year angiographic restenosis rate of 2%. Patients selected for CAS may represent a subset of patients with carotid disease who have considerable comorbidities or unfavorable anatomy compared to those undergoing conventional endarterectomy. CAS may be performed safely outside of a clinical trial with results similar to those of published series from trial centers using a standardized protocol. PMID- 14534846 TI - Percutaneous therapy for renal artery fibromuscular dysplasia. AB - Fibromuscular dysplasia (FMD) accounts for 10% of cases of renal artery stenosis. This study evaluates the anatomic and functional outcomes of endovascular therapy for symptomatic renal artery FMD at an academic medical center. A retrospective analysis of records from patients who underwent percutaneous transluminal renal artery angioplasty (PTRA) found 14 patients (all female) who underwent 19 interventions on 18 renal artery segments. Significant cardiovascular comorbidities were few in this patient population. The PTRA technical success rate was 95%. There were no periprocedural mortalities. Primary patency rates were 81%, 69%, 69%, and 69% at 2, 4, 6, and 8 years. Assisted primary patency rates were 87%, 87%, 87%, and 87% at 2, 4, 6, and 8 years. The restenosis rate was 25% at 8 years. Clinical benefit (improved or cured hypertension) was seen in 79% of patients overall; 65% of patients maintained this benefit at 8 years by life-table analysis. Percutaneous endovascular intervention for clinically symptomatic FMD of the renal arteries is technically successful, safe, and durable. Most patients demonstrate immediate clinical benefit and retain durable functional outcomes. PMID- 14534847 TI - Re-evaluation of carotid duplex for visual complaints: who really needs to be studied? AB - Amaurosis fugax (AF), Hollenhorst plaques, central retinal artery occlusion (CRAO), and nonspecific visual symptoms are all reasons for patient referral for carotid artery evaluation. This study reviews the management of patients with visual signs or symptoms based on their clinical presentation, carotid duplex results, follow-up data, and outcome. We performed a retrospective review of all patients presenting to the Vascular Surgery Clinic between June 1996 and December 2001 for carotid duplex scanning because of the indication of a visual disturbance. A total of 3560 carotid duplex examinations were performed during the study period; 98 were performed for a visual complaint or finding. A total of 11.1% of group 1 (Hollenhorst plaques), 22.2% in group 2 (CRAO), 45% in group 3 (AF), and 9.8% in group 4 (nonspecific visual symptoms) had significant carotid disease and underwent carotid endarterectomy. No patient who underwent screening carotid duplex and did not have surgically correctable disease developed significant carotid disease or symptoms from carotid disease during the study period. Hollenhorst plaques, CRAO, and nonspecific visual complaints are a poor predictor of significant carotid stenosis, while AF had a significantly higher rate of surgically correctable carotid stenosis. Patients with visual signs or symptoms need an initial screening carotid duplex examination. If this does not show surgically correctable disease, patients do not need to return for further examinations unless another indication arises. PMID- 14534848 TI - Vascular injuries associated with elective orthopedic procedures. AB - The objective of this study was to review the diagnosis, management, and outcome of the rare iatrogenic arterial injury associated with elective orthopedic joint procedures. A retrospective review was conducted of all patients presenting to the vascular surgery service with arterial injury after elective orthopedic procedures between 1997 and 2002. Clinical records were reviewed for presentation, type of injury, management, and outcome. During the study period, 20 patients having 21 total orthopedic procedures were identified with 27 arterial injuries. There were 4350 elective orthopedic procedures during this period for an incidence of 0.005%. There were 14 total knee arthroplasties, 4 total hip arthroplasties, and 3 ankle reconstructions in the study group. Presenting signs included acute ischemia with loss-of-limb Doppler-detected arterial flow/pulses (13 patients, 62%), intraoperative arterial bleeding (3 patients, 14%), nonhealing wounds (3 patients, 14%), and limb edema (2 patients, 10%); the diagnosis was delayed >24 hr in 5 patients (25%). Arterial thrombosis was the most common abnormality identified (21 of 27 injuries, 78%), followed by laceration/avulsion (3 injuries, 11%) and pseudoaneurysm development (3 patients, 11%), and involved the iliac ( n = 3), common femoral ( n = 2), profunda ( n = 1), superficial femoral ( n = 4), popliteal ( n = 12), or tibial ( n = 5) arteries. Concomitant popliteal venous injury was present in one patient. Injured arterial segments had preexisting atherosclerotic disease (33%) and 15 patients (71%) had prior surgery in proximity to the arterial injury while an additional 9 (43%) had prior traumatic injury (7 [78%] of whom had revision orthopedic surgery as well). Management consisted of vein bypass grafting ( n = 15, 56%), primary repair ( n = 3, 11%), and thrombectomy with thrombolysis ( n = 2, 7%). One patient (5%) underwent primary above-knee amputation. There was one death from septic shock and there were three limb losses (14%). Arterial injury associated with elective orthopedic joint surgery is more common during redoprocedures and in patients with preexisting atherosclerosis. Despite arterial repair/bypass, limb morbidity is common and related to preexisting occlusive disease or extent of arterial thrombosis. PMID- 14534850 TI - Characterization and spatial distribution of ectomycorrhizas colonizing aspen clones released in an experimental field. AB - Ectomycorrhizas (EM) from aspen clones released on an experimental field were characterized by morphotyping, restriction analysis and internal transcribed spacer (ITS) sequencing. In addition, their community structure and spatial distribution was analyzed. Among the 23 observed morphotypes, six mycobionts dominated, forming roughly 90% of all ectomycorrhizas: Cenococcum geophilum, Laccaria sp., Phialocephala fortinii, two different Thelephoraceae, and one member of the Pezizales. The three most common morphotypes had an even spatial distribution, reflecting the high degree of homogeneity of the experimental field. The distribution of three other morphotypes was correlated with the distances to the spruce forest and deciduous trees bordering the experimental field. These two patterns allowed two invasion strategies of ectomycorrhizal fungi (EMF) to be recognized, the success of which depends on adaptation of the EMF to local ecological conditions. PMID- 14534849 TI - The aging spine: new technologies and therapeutics for the osteoporotic spine. AB - Osteoporosis results in low-energy fractures of the spine. The load necessary to cause a vertebral fracture is determined by the characteristics related to the vertebral body structure, mineral content, and quality of bone. Radiographic techniques centered on dual X-ray absorptiometry (DXA) permit a determination of bone mass and fracture risk. Current medical therapies principally using bisphosphonate and pulsatile PTH profoundly decrease the risk of fracture (50+%). Fall prevention strategies can further decrease the possibility of fracture. A comprehensive approach to osteoporosis can favorably alter the disease. PMID- 14534851 TI - The population of the hypogeous fungus Tuber aestivum syn. T. uncinatum on the island of Gotland. AB - The aim of our study was to examine the genetic variation within Tuber aestivum on the Baltic island of Gotland, Sweden. Variation in such a limited geographical area should help illuminate the dispersal abilities of T. aestivum. Knowledge of the genetic variation in this northern outpost could also be useful in the selection of inoculum for the establishment of truffle orchards. Genetic structure and homogeneity of the population were studied using principal component and parsimony analyses of randomly amplified polymorphic DNA data. Our inventories showed that T. aestivum is abundantly distributed in suitable habitats on Gotland. The genetic variation observed suggests sexual reproduction and slow dispersal on the island. It is possible that the present population was established from one introduction, which may be due to ability to survive in this habitat rather than to rare colonising events. The T. aestivum population on Gotland may be an ecotype adapted to the climate and soil conditions on the island. PMID- 14534853 TI - Emerging immunomodulatory therapies targeting the co-stimulatory pathways for the prevention of transplant rejection. AB - The continued discovery of novel immune molecules and pathways over the last decade has spurred a tremendous amount of research into the targeting of these pathways for the prevention of transplant rejection. Of particular interest are members of the ever-growing family of T-cell co-stimulatory pathways; the classic co-stimulatory pathways are the CD28/CTLA4:B7-1/2 and CD40:CD154, while the ICOS:ICOSL and PD-1:PD-L1/PD-L2 pathways are novel. Various chimeric molecules and monoclonal antibodies have been developed for targeting these pathways with promising results, especially with the newer generation of agents and in combination among themselves, with other immunomodulatory therapies and/or with conventional immunosuppressive agents. These novel immunomodulatory strategies have the potential to bring us a step closer to achieving transplantation tolerance. PMID- 14534852 TI - Preoperative factors predictive of complicated postoperative management after Roux-en-Y gastric bypass for morbid obesity. AB - INTRODUCTION: This study was undertaken to determine preoperative predictive factors of complicated postoperative management after Roux-en-Y gastric bypass (RYGB) for morbid obesity. METHODS: Between January 1999 and January 2002, 158 patients who underwent a RYGB received a standardized preoperative evaluation and data were collected prospectively. Complicated postoperative management was defined as patients requiring postoperative ICU admission for > or =48 h, or those needing transfer from the floor to the ICU. Patients with complicated management were compared with those in whom ICU admission was not necessary. RESULTS: Twenty-three patients (14.5%) required prolonged ICU admission (mean stay of 6.3 +/- 1.7 days). After multivariate analysis, body mass index (BMI) >50 kg/m2, forced expiratory volume (FEV1) <80% predicted, previous abdominal surgeries, and abnormal EKG were found to be independently associated with an increased likelihood of complicated postoperative care. CONCLUSION: BMI >50 kg/m2, FEV1 <80% predicted, previous abdominal surgeries, and abnormal EKG increase the likelihood of complicated postoperative management after RYGB for morbid obesity. PMID- 14534854 TI - Bradykinin receptor antagonists in cerebral ischemia and trauma. AB - Bradykinin, an endogenous nonapeptide produced by activation of the kallikrein kinin system, promotes neuronal tissue damage as well as disturbances in blood brain barrier function, mainly through the activation of B2 receptors. Both cerebral ischemia and trauma are associated with an activation of the kallikrein kinin system, resulting in a sustained production of kinins. This is strong evidence supporting an involvement of bradykinin-mediated secondary brain damage following cerebral ischemia and trauma. Accordingly, specific inhibition of bradykinin B2 receptors attenuates brain swelling and limits tissue damage, indicating its potential in patients with acute cerebral ischemia and trauma. PMID- 14534855 TI - Dexanabinol: a novel cannabinoid with neuroprotective properties. AB - Dexanabinol was developed as a synthetic analog of the cannabinoid tetrahydrocannabinol, however, the action of the drug is as a weak, uncompetitive antagonist at the N-methyl-D-aspartate (NMDA) receptor. Dexanabinol is an effective neuroprotectant in animal models of hypoxemia/ischemia, neurotoxin exposure and nerve crush. Unlike some other uncompetitive NMDA receptor antagonists, dexanabinol does not produce psychotrophic effects and is generally well tolerated in humans. In this respect it is similar to the uncompetitive NMDA receptor antagonist memantine. Dexanabinol is currently in phase III clinical trials for severe head injury. It is a promising drug that appears to be able to provide the therapeutic benefits of uncompetitive NMDA-receptor antagonists without the adverse side effects associated with some of the earlier drugs of this type. PMID- 14534856 TI - Anidulafungin Vicuron. AB - Anidulafungin, a 1,3-beta-glucan synthesis inhibitor and the pentyloxyterphenyl side chain derivative of echinocandin B, is being developed by Vicuron Pharmaceuticals Inc, under license from Eli Lilly & Co, for the potential treatment of fungal infection. PMID- 14534857 TI - Ferumoxtran-10 Advanced Magnetics. AB - Ferumoxtran-10 (Combidex) is an ultra-small superparamagnetic iron oxide molecular resonance imaging contrast agent under development by Advanced Magnetics Ltd and Guerbet for the principal indication of lymph node imaging. PMID- 14534858 TI - [Evidence and advances in cardiology]. PMID- 14534859 TI - [Dentogenic foci and non-evidence based restorative methods]. PMID- 14534860 TI - [Too many heart catheter procedures in Germany ?]. AB - BACKGROUND AND OBJECTIVE: Absolute numbers of cardiovascular procedures are higher in Germany as compared to other European countries. This fact is used as an argument for overuse. Therefore other indicators of an inappropriate use of these resources should be of interest. PATIENTS AND METHODS: The relationship between diagnostic cardiac catheterisations and consequent revascularisation procedures were compared in 8 European countries. In addition the indication criteria for cardiac catheterisations were reviewed in a German registry of 205.581 consecutive inpatients. RESULTS: Revascularisation procedures after diagnostic catheterisations in 8 countries range from 39,1 % to 57,9 %. Germany reaches 43,2 %. A relation between absolute numbers of diagnostic and percent subsequent revascularisation procedures does not exist. In a German registry the following indications for cardiac catheterisation could be identified: Acute Coronary Syndrome 22,9 %. Angina pectoris according to the Canadian Cardiac Society classification was present: CCS II/III in 80,3 %, CCS IV in 17,2 %. An exercise test was performed in 43 %. Final diagnoses were: significant coronary disease 69,5 %, exclusion of disease 9,4 %, lesions < 50 % 9 %, other cardiac disease 12,1 %. CONCLUSION: Absolute numbers cannot be used as an indicator of overuse of cardiovascular procedures. Instead standards for data acquisition should be established on European, national and regional levels. In addition a validation procedure for criteria has to be developed in order to judge the appropriateness of indications for invasive cardiac procedures in different health care systems. PMID- 14534861 TI - [Clinical features and current treatment of hypertensive crisis]. PMID- 14534862 TI - [Topical questions regarding off-label use]. PMID- 14534863 TI - [Aortic valve insufficiency: when is the optimal moment for surgery?]. PMID- 14534864 TI - [Optimal thrombolytic strategies for acute myocardial infarction: nevertheless opting for percutaneous intervention (PCI) ?]. PMID- 14534865 TI - [Diagnosis of ischemia--adequate clinical and exercise-ECG for indicating the need for cardiac catheterization?--Pro]. PMID- 14534866 TI - [Diagnosis of ischemia--adequate clinical and exercise-ECG for indicating the need for cardiac catheterization?--Contra]. PMID- 14534867 TI - [If the spirit of the heart agonizes. Nervous heartbeats, First World War and the popularization of cardiac neurosis]. PMID- 14534868 TI - What modern physicians can learn from Hippocrates. PMID- 14534869 TI - High time for low-dose prospective clinical trials. PMID- 14534870 TI - Application of immunofluorescence for intraoperative evaluation of sentinel lymph nodes in patients with breast carcinoma. AB - BACKGROUND: Sentinel lymph node (SLN) sampling is used to predict axillary lymph node (AxLN) metastasis in patients with breast carcinoma; AxLN sampling can be avoided in SLN-negative patients. Multistep sectioning and immunohistochemistry are reported to increase the sensitivity of micrometastasis detection. The authors used immunofluorescence (IF) staining with cytokeratin (CK) antibodies for intraoperative evaluation of touch imprints from multistep sections of SLNs. METHODS: A combined total of 150 SLNs from 69 patients who had breast carcinoma and underwent surgery were analyzed. SLNs were sectioned at 2 mm intervals, and touch imprints subsequently were prepared. After ethanol fixation, slides were incubated with fluorescein isothiocyanate-conjugated CK and propidium iodide and then subjected to microwave irradiation for 3 minutes. Finally, fluorescence microscopy was used to evaluate slides for the presence or absence of metastatic disease. RESULTS: Metastatic disease was identified in 36 (24.0%), 26 (17.3%), 32 (21.3%), and 31 (20.7%) of 150 lymph nodes using IF staining, Papanicolaou (Pap) staining, immunocytochemical (ICC) staining, and hematoxylin and eosin (H and E) staining, respectively. The sensitivities of IF, Pap, ICC, and H and E staining were 94.6%, 70.3%, 86.5%, and 83.8%, respectively. Except for IF staining (specificity, 99.1%), each method had a specificity of 100%. The accuracy rates for IF, Pap, ICC, and H and E staining were 98.0%, 92.7%, 96.7%, and 96.0%, respectively. CONCLUSIONS: IF is a rapid, highly sensitive, and highly specific staining technique by which touch imprints can be used to intraoperatively evaluate SLNs in patients with breast carcinoma. PMID- 14534871 TI - Disruption of the expected positive correlation between breast tumor size and lymph node status in BRCA1-related breast carcinoma. AB - BACKGROUND: A positive correlation between breast tumor size and the number of axillary lymph nodes containing tumor is well established. It has been reported that patients with BRCA1-related breast carcinoma are more likely than patients with nonhereditary breast carcinoma to have negative lymph node status. Therefore, the authors questioned whether the known positive correlation between tumor size and lymph node status also was present in women with BRCA1-related breast carcinomas. METHODS: The relation between the greatest dimension of the resected breast tumor (size) and the presence of positive axillary lymph nodes (expressed as a percentage of all lymph nodes examined) was evaluated in 1555 women with invasive breast carcinoma who were ascertained at 10 centers in North America between 1975 and 1997. There were 276 BRCA1 mutation carriers, 136 BRCA2 carriers, and 1143 women without a known mutation (208 BRCA1/BRCA2 noncarriers and 935 untested women). Patients were stratified according to tumor size, and odds ratios were estimated for the presence of positive lymph nodes with increasing tumor size. RESULTS: A highly significant positive correlation between tumor size and the frequency of positive axillary lymph nodes was seen for BRCA1/BRCA2 noncarriers, for BRCA2 carriers, and for untested women (overall P < 0.0001 for each). In contrast, there was no clear correlation between tumor size and positive lymph node status in BRCA1 carriers (overall P = 0.20). CONCLUSIONS: The relation between tumor size and lymph node status in patients with breast carcinoma appears to be different in BRCA1 carriers compared with BRCA2 carriers and noncarriers. These findings have important implications for estimating the route of metastatic spread and for evaluating the effectiveness of early diagnosis in patients with BRCA1-related breast carcinoma. PMID- 14534872 TI - Coagulopathic complications in breast cancer. AB - Patients with cancer are highly susceptible to thromboembolic complications, which account for a significant percentage of the morbidity and mortality of the disease. Up to 15% of patients with clinically overt cancer present with venous thromboembolism during the course of their disease. Moreover, patients with cancer represent 20% of all patients in whom deep venous thrombosis and pulmonary embolism are diagnosed. This prothrombotic state in cancer can occur due to the ability of tumor cells to directly activate the blood-clotting cascade and cause thrombosis or induce procoagulant properties and inhibit anticoagulant properties of vascular endothelial cells, platelets, monocytes, and macrophages. It also is well established that this prothrombotic tendency in patients with cancer can be enhanced greatly by anticancer treatments, such as surgery and chemotherapy. This phenomenon can be seen in patients with breast cancer, particularly after surgery and chemotherapy. Increased clotting risk also is associated with the use of central venous access devices, commonly used to administer chemotherapeutic agents in patients with cancer. Thrombosis prophylaxis, therefore, should be considered for patients with breast cancer who are at risk before and during intervention. In the current review, the authors discuss the problem of thromboembolism in patients with breast cancer who are undergoing therapy, the mechanisms by which thromboembolisms occur, and the potential strategies by which these events may be prevented. Better understanding of these pathogenetic pathways may lead to the development of more targeted strategies to prevent thromboembolism in patients with cancer. PMID- 14534874 TI - Relation between the method of detection of initial breast carcinoma and the method of detection of subsequent ipsilateral local recurrence and contralateral breast carcinoma. AB - BACKGROUND: Mammography and physical examination are routine methods for the detection of ipsilateral local recurrence and contralateral breast carcinoma in patients initially undergoing breast conservation treatment. The current study reports the relation between the method of detection of the initial breast carcinoma and the method of detection of subsequent ipsilateral local recurrence and contralateral breast carcinoma. METHODS: A retrospective review was performed of the records of female patients with initial American Joint Committee on Cancer (AJCC) Stage I and II invasive breast carcinoma who developed ipsilateral local recurrence or contralateral breast carcinoma after breast conservation treatment. The method of detection of local recurrence in the ipsilateral breast and the method of detection of contralateral breast carcinoma were compared with the method of detection of the primary tumor. RESULTS: There were 125 ipsilateral breast local recurrences and 71 contralateral breast carcinoma cases detected. Of the 125 recurrences in the ipsilateral breast, 38% (48 recurrences) were detected by mammography only, 37% (46 recurrences) were detected by physical examination only, and 25% (31 recurrences) were detected by both methods. Of the 71 contralateral breast carcinoma cases, 53% (38 cases) were detected by mammography only, 23% (16 cases) were detected by physical examination only, and 24% (17 cases) were detected by both methods. When the primary tumors were detected by mammography only, 21% of the local recurrences (3 of 14 local recurrences) and 19% of the contralateral breast carcinoma cases (4 of 21 cases) were detected by physical examination only. When the primary tumors were detected by physical examination only, 24% of the local recurrences (14 of 58 local recurrences) and 42% of the contralateral breast carcinoma cases (8 of 19 cases) were detected by mammography only. When stratified by the interval between diagnosis of the primary tumor and ipsilateral local recurrence or contralateral breast carcinoma (< or = 5 years vs. > 5 years) or age of the patient at the time of ipsilateral breast recurrence or contralateral breast carcinoma (age < or = 49 years vs. age > or = 50 years), both breast examination and mammography were found to be important in the detection of locally recurrent tumor and contralateral breast carcinoma in each subgroup of the patients, regardless of the method of presentation of the primary tumor. CONCLUSIONS: Both mammography and physical examination were found to be significant in the detection of locally recurrent tumor in the ipsilateral breast and in the detection of contralateral breast carcinoma, regardless of the method of detection of the primary tumor. PMID- 14534873 TI - A comparison of five immunohistochemical biomarkers and HER-2/neu gene amplification by fluorescence in situ hybridization in white and Korean patients with early-onset breast carcinoma. AB - BACKGROUND: The objective of this article was to compare five tumor markers between white women in the U.S. and native Korean women with early-onset breast carcinoma. METHODS: Sixty Korean women who were diagnosed with breast carcinoma at age 45 years or younger and 60 white women with breast carcinoma who were matched by age were selected for this study. The median age of both groups was 37 years. Paraffin embedded blocks of the primary tumor were processed for immunohistochemical staining of estrogen receptor (ER), progesterone receptor (PR), p53, cyclin D1, and HER-2/neu. RESULTS: The proportion of tumors that stained positive for ER, PR, p53, and cyclin D1 in the Korean women were 47.5%, 42.4%, 28.8%, and 40.9%, respectively; in the white women, the proportions were 43.9%, 52.6%, 21.1%, and 59.1%, respectively. The differences between the white patients and the Korean patients were not statistically significant with respect to any of those variables. A significant difference was found in the expression of HER-2/neu. Specifically, positive HER-2/neu status was observed in 47.5% of Korean women, compared with overexpression in only 15.8% of white women (P < 0.001). Fluorescence in situ hybridization analysis for HER-2/neu gene amplification on all HER-2/neu positive samples that scored 2 + and 3 + demonstrated a significant difference (P = 0.007) in gene amplification between the two populations. Differences in HER-2/neu positivity were observed for the entire cohort as well as among the subsets of patients with negative and positive lymph node status. No association was found between immunoreactivity for the five markers and axillary lymph node metastasis. CONCLUSIONS: The findings of high positivity of HER-2/neu expression and gene amplification in Korean women with early-onset breast carcinoma may have potential implications for local and systemic management of breast carcinoma, especially anti-HER-2/neu therapy for patients with hormone receptor negativity. Further research will be needed to identify biologic and genetic factors and their effects on the survival between different racial groups. PMID- 14534875 TI - Phase II trial of oral cyclophosphamide, prednisone, and diethylstilbestrol for androgen-independent prostate carcinoma. AB - BACKGROUND: The authors evaluated the combination of oral cyclophosphamide, oral prednisone, and diethylstilbestrol (DES) in patients with androgen-independent prostate carcinoma (AIPC). METHODS: Thirty-seven patients with prostate carcinoma refractory to androgen ablation who had undergone antiandrogen withdrawal (if previously treated with an antiandrogen) were enrolled in the current study. They were treated with oral cyclophosphamide 100 mg per day on Days 1-20, prednisone 10 mg per day continuously, and DES 1 mg continuously, on a 30-day cycle. Warfarin 1 mg per day was given as prophylaxis for thrombosis. Patient levels of prostate-specific antigen (PSA) were monitored on a monthly basis, with imaging studies every 3 months. Patients continued to receive therapy until disease progression or the occurrence of significant toxicity. The effect of therapy on the patient's quality of life was assessed using the Functional Assessment of Cancer Therapy-Prostate. RESULTS: Thirty-six patients were evaluable for response. Of the 36 patients, 15 (42%) had a 50% or greater decline in PSA levels from pretreatment levels and 1 patient (6%) with measurable disease had a partial response to therapy. The median duration of response was 4.5 months (range, 4-18 months). The overall median survival period was 16.4 months. The treatment was well tolerated, with only three patients removed from the study for toxicities associated with treatment. One patient, who had been treated for more than 24 months, developed acute leukemia. Quality of life evaluation in 17 patients showed a significant improvement in responders, whereas nonresponders had no deterioration while receiving therapy. CONCLUSIONS: Cyclophosphamide, prednisone, and DES represent a well tolerated, low-cost combination therapy with significant activity in the treatment of patients with AIPC. PMID- 14534876 TI - Response assessment classification in patients with advanced renal cell carcinoma treated on clinical trials. AB - BACKGROUND: The objective of the current study was to evaluate the effect on response assessment classification in patients with metastatic renal cell carcinoma (RCC) using unidimensional (Response Evaluation Criteria in Solid Tumors) and bidimensional (World Health Organization) criteria, including or excluding measurements of the primary renal tumor and using a new index to compensate for the disproportionate effect of large renal tumors relative to their metastases. METHODS: Fifty-three imaging studies involving a total of 44 patients with metastatic RCC who were treated on clinical trials of interferon alpha analogue and/or thalidomide were reviewed retrospectively. The best overall response assessment and progression free survival were calculated with both unidimensional and bidimensional tumor measurements. Patients were then stratified into two groups: patients with primary renal tumors in situ and patients who underwent resection of their primary renal tumors. The best overall response and the time to disease progression were calculated based on the sum of measurements (conventional methodology), both including and excluding the primary tumor. A new method of response assessment, the 'normalized lesion index', which equalizes the differences in tumor size for an individual patient, was evaluated and compared with the conventional response assessment. RESULTS: There was an 11% disagreement rate in the best overall response assessment between unidimensional and bidimensional measurements. The time to progression was 9.2 months measured unidimensionally, compared with 6.4 months assessed bidimensionally. In the group of patients who had primary renal tumors in situ, using the conventional sum of measurements method, the apparent time to progression was an average of 4.2 months longer compared with measurements that did not include the primary renal tumor. The use of the normalized lesion index method resulted in an improved concordance in best overall response assessments and similar time to progression assessments when the primary renal tumor was included compared with patients who did not have primary renal tumors in situ. CONCLUSIONS: The use of unidimensional measurements in RCC therapy assessment results in significantly different time to progression classification compared with the use of bidimensional measurements. Response assessment classification in patients with RCC is affected by the exclusion or inclusion of measurements of the primary renal tumor. The normalized lesion index warrants further study in assessing response in patients with metastatic RCC and other solid tumor malignancies that often show substantial differences in sizes of measurable lesions. PMID- 14534877 TI - The development of bladder tumors and contralateral upper urinary tract tumors after primary transitional cell carcinoma of the upper urinary tract. AB - BACKGROUND: Contralateral, metachronous upper urinary tract (UUT) tumors after primary transitional cell carcinoma (TCC) of the UUT are reported rarely, and to the authors' knowledge the risk factors have not been determined to date. In addition, few reports have described the characteristics of recurrent bladder tumors and contralateral UUT tumors and any relation between theses tumor types. METHODS: Statistical analysis of data from 223 patients with documented primary UUT-TCC was undertaken. After excluding bilateral involvement and distant metastases, 12 variables were analyzed by multivariate analysis in 189 patients to determine the risk factors for recurrent urothelial tumors. RESULTS: The incidence rates of recurrent bladder tumors and contralateral UUT tumors were 31.2% and 5.8%, respectively. Multiplicity was determined as a risk factor for recurrent bladder tumors. Renal insufficiency, uremia, and concurrent bladder tumors significantly predisposed patients to develop contralateral UUT tumors after primary UUT-TCC. The time intervals and stage distributions differed significantly between recurrent bladder tumors and contralateral UTT tumors. Patients who had recurrent bladder tumors had earlier stage tumors and had a shorter time to recur compared with patients who had contralateral, metachronous UUT tumors. CONCLUSIONS: For patients with primary UUT-TCC, regular follow-up by cystoscopy is necessary to detect recurrent bladder tumors. Intravenous urography or retrograde pyelography should be performed for patients who have a high risk of developing contralateral UUT tumors. PMID- 14534878 TI - Weekly administration of docetaxel for symptomatic metastatic hormone-refractory prostate carcinoma. AB - BACKGROUND: The current Phase II study investigated the clinical benefit, impact on quality of life (QOL), and tolerability of weekly docetaxel in symptomatic patients with metastatic hormone-refractory prostate carcinoma (HRPC). METHODS: Patients received weekly docetaxel 35 mg/m(2) intravenously for 6 consecutive weeks followed by a 2-week rest repeatedly for a maximum of 24 weeks of treatment. Clinical benefit evaluations, based on Karnofsky performance status (KPS) and pain, were assessed weekly during therapy. A clinical benefit response was defined as a sustained (> or = 4-week) improvement in at least one of these parameters without worsening in the other. Patient-assessed QOL (using the European Organization for Research and Treatment of Cancer QLQ-C30), changes in prostate-specific antigen (PSA) levels, tumoral response, and toxicity also were evaluated. RESULTS: Thirty men (median age, 67 years), 15 of whom had received previous chemotherapy, were treated. Overall, 46% of patients achieved a positive pain response and 48% achieved a 50%-or-greater reduction in PSA. KPS was high at baseline (80%), and no significant changes in this parameter were observed. Compared with baseline, all scores improved after the first cycle of therapy, particularly emotional (P = 0.015), pain (P = 0.001), constipation (P = 0.001), and global QOL (P = 0.011) scores. After the second cycle, dyspnea scores decreased (P = 0.010). At the last QOL assessment, there also was deterioration in terms of fatigue (P = 0.013), dyspnea (P = 0.010), and physical functioning (P = 0.017). Toxicity was mild and included neutropenia (Grade 3-4, n = 2). CONCLUSIONS: Of these elderly symptomatic patients with HRPC, half had received previous chemotherapy. Weekly docetaxel was found to be associated with improvements in clinical benefit response and in QOL and was well tolerated. PMID- 14534879 TI - Factors that influence the results of salvage surgery in patients with chemorefractory germ cell carcinomas with elevated tumor markers. AB - BACKGROUND: A standard concept for the integration of surgery into the chemotherapy-based treatment of patients with advanced germ cell carcinoma has been that surgery should be avoided in patients with serum tumor markers (alpha fetoprotein [AFP] and human chorionic gonadotropin [HCG]) that remain elevated. However, some patients may benefit from resection under such chemorefractory conditions. The objective of this retrospective study was to clarify the outcome and clinical prognostic variables of salvage surgery in patients with disseminated (AJCC Stage II or III) testicular germ cell carcinoma or extragonadal germ cell carcinoma who had elevated serum markers. METHODS: The authors reviewed the clinical records of 24 patients who underwent salvage surgery with elevated serum AFP and/or HCG levels after at least 3 courses of cisplatin-based, systemic chemotherapy between January, 1985 and December, 2000. The survival rates were compared between the subgroups with regard to preoperative and postoperative clinical parameters using the Kaplan-Meier method and a Cox proportional hazards model. RESULTS: Ten of 24 patients (41.7%) were rendered free of disease and alive without disease after the surgery with or without adjuvant therapy at a median follow-up of 74 months (range, 24-207 months). Among the preoperative parameters, high HCG levels were associated with significantly poorer survival (hazard ratio [HR], 8.321; 95% confidence interval [95% CI], 1.0753-64.553; P = 0.043 and P = 0.016, respectively; log-rank test). In addition, patients who had visceral lesions at resection had a significantly poorer prognosis compared with patients who had retroperitoneal and/or mediastinal lymph node lesions (P = 0.038; log-rank test). Among postoperative parameters, incomplete resection and persistently high HCG levels were associated significantly with poor survival, with a risk of death from disease of 12.516 fold (95% CI, 1.786-87.781) and 9.311-fold (95% CI, 1.796-48.256), respectively. CONCLUSIONS: Salvage surgery in patients with high serum tumors markers resulted in long-term disease free status in approximately 40% of patients in a small subset with advanced germ cell carcinoma. Patients with elevated AFP levels alone (i.e., normal HCG levels) or with lymph node lesions alone seem to be good candidates for such surgery. Complete resection of target lesions and normalization of HCG levels after surgery are mandatory to achieve long-term disease free status. PMID- 14534880 TI - Metronomic therapy with cyclophosphamide and dexamethasone for prostate carcinoma. AB - BACKGROUND: The current study was designed to evaluate the efficacy and toxicity of the continuous oral administration of a combination of cyclophosphamide (50 mg/day given in the morning) and dexamethasone (1 mg/day given in the evening) in patients with prostate specific antigen (PSA) progression despite single or multiagent hormone therapy and antiandrogen withdrawal. METHODS: The authors retrospectively evaluated the medical records of all patients with prostate carcinoma who were treated with dexamethasone and cyclophosphamide and who were unable to participate in Phase II drug trials or had failed previous chemotherapy regimens. RESULTS: Using clinical response guidelines set forth by the Prostate Specific Antigen Working Group, 29% of patients were found to have a > or = 80% reduction in PSA, 39% were found to have a 50-79% reduction in PSA, 6% were found to have a < 50% decrease in PSA, and 26% experienced disease progression while receiving treatment. The duration of response was 8 months (95% confidence interval [95% CI], 4-10 months). The duration of treatment was 9 months (95% CI, 6-14 months). The treatment was reported to be well tolerated with side effects being primarily bruising, Cushingoid facies, and gastrointestinal distress. CONCLUSIONS: In the current study, low-dose dexamethasone and cyclophosphamide demonstrated efficacy as salvage therapy in the treatment of patients with hormone-refractory prostate carcinoma. PMID- 14534881 TI - Motility-related protein 1 (MRP-1/CD9) expression in urothelial bladder carcinoma and its relation to tumor recurrence and progression. AB - BACKGROUND: CD9 has been implicated in cell adhesion, motility, and proliferation, and numerous studies have demonstrated its prognostic value in different solid tumors. The objective of this study was to determine the relation of CD9 expression to tumor grade and tumor stage of urothelial carcinoma of the bladder and to define the value of CD9 in predicting the behavior of superficial papillary tumors (SPTs) (pathologic Ta [pTa] and pT1). METHODS: Three hundred twenty patients (118 patients with pTa tumors, 111 patients with pT1 tumors, and 91 patients with pT2 tumors) were examined for CD9 expression using immunohistochemistry applied on formalin fixed, paraffin embedded tissue. Patients were stratified into 3 categories, depending on CD9 expression: positive (> 50% positive cells), reduced (5-50% positive cells), or negative (< 5% positive cells). RESULTS: Loss of CD9 expression was found to be associated significantly with high-grade and high-stage urothelial tumors (P < 0.0001). A reduced/negative (altered) CD9 expression was associated with SPT progression, but not with recurrence (P < 0.001). Patients who had pTa or pT1 tumors with altered CD9 expression had a relative risk of 5.59 (P = 0.005; 95% confidence interval [95% CI], 1.69-18.48) for progression compared with patients who had tumors with positive CD9 expression. Kaplan-Meier curves showed that a lack of CD9 expression was associated significantly with progression free survival (P < 0.001; log-rank test), but not with recurrence. In patients with SPTs, multivariate Cox proportional hazards regression analysis revealed that negative CD9 expression was an independent prognostic marker for the prediction of tumor progression (P = 0.007; 95% CI, 0.11-0.70). CONCLUSIONS: In patients with urothelial bladder carcinoma, CD9 expression was associated significantly with tumor stage and grade, and a loss of CD9 expression was an independent prognostic factor for predicting progression in patients with SPTs. Thus, CD9 immunoexpression is a potential new predictor of tumor behavior in patients with SPTs of the urinary bladder. PMID- 14534882 TI - Endometriosis-associated ovarian carcinoma: differential expression of vascular endothelial growth factor and estrogen/progesterone receptors. AB - BACKGROUND: Multiple epidemiologic and histologic studies have suggested that ovarian endometriosis can give rise to malignant ovarian tumors, primarily those of epithelial origin. The progression of endometriosis to endometriosis associated ovarian carcinoma (EAOC) has not been investigated thoroughly and is poorly understood at best. Using immunohistochemical methods, we compared the differential expression patterns of various cytokines and growth factors in atypical endometriosis (AE) and EAOC. METHODS: Using the Johns Hopkins Pathology Data Bank, tissue blocks from patients diagnosed with EAOC or AE were identified. Tissue blocks were stained for 4 markers: vascular endothelial growth factor (VEGF), Ki-67, estrogen receptor (ER), and progesterone receptor (PR). RESULTS: Seventeen cases of EAOC and 8 cases of AE were identified. Staining for VEGF was documented in 16 of 17 (94%) EAOC tissue blocks and in only 1 of 8 (12.5%) AE tissue blocks (P < 0.0001). Only 4 of the 17 (23%) EAOC tissue blocks exhibited positive staining for ER, compared with 8 of 8 (100%) AE tissue blocks (P = 0.0005). Positive staining for PR was noted in only 6 of 17 (35%) EAOC samples but was present in 8 of 8 (100%) AE samples (P = 0.003). Seventy percent of EAOC samples exhibited positive staining for Ki-67, compared with 37.5% of AE samples (P = 0.19). CONCLUSIONS: EAOC appears to be associated with overexpression of VEGF and reduced expression of both ER and PR. Variations in VEGF expression may be associated with the malignant transformation of endometriosis and may present both diagnostic and therapeutic options for the treatment of ovarian malignancies. PMID- 14534883 TI - Phase II evaluation of three-day topotecan in recurrent platinum-sensitive ovarian carcinoma: a gynecologic oncology group study. AB - BACKGROUND: Topotecan, administered intravenously at a dose of 1.5 mg/m(2) per day for 5 days every 21 days, is an established regimen in the treatment of recurrent ovarian carcinoma. Alternate dosing strategies have sought to improve toxicity. The authors evaluated the tolerability and antitumor activity of a 3 day topotecan regimen. METHODS: A multicenter Phase II study, which included patients with platinum-sensitive ovarian carcinoma, was conducted. Patients were to receive an intravenous dose of topotecan of 2.0 mg/m(2) per day for 3 days every 21 days until disease progression or unacceptable toxicity occurred. Doses were modified in 0.25-mg/m(2) increments based on tolerability. Granulocyte colony-stimulating factor support was used as necessary. RESULTS: From February to June 2000, 30 patients were enrolled. Their median age was 56 years (range, 41 81 years). Twenty-nine patients were evaluable for toxicity and efficacy. A median of 5 courses (range, 1-11 courses) of topotecan was administered. Eighteen of 30 (60%) patients experienced Grade 4 neutropenia. There was one report each of Grade 4 thrombocytopenia, anemia, and gastrointestinal toxicity (grading performed according to National Cancer Institute Common Toxicity Criteria). Ten patients developed Grade 3 leukopenia and 9 had Grade 3 neutropenia. Serious nonhematologic events were rare. There were 2 (7%) complete and 2 (7%) partial responses, for an overall response rate of 14%. Sixteen (55%) patients had stable disease and 9 (31%) experienced disease progression. CONCLUSIONS: A 3-day regimen of topotecan at a dose of 2.0 mg/m(2) per day was generally well tolerated, although the response rate was lower than that for the standard 5-day schedule. PMID- 14534884 TI - National Cancer Data Base report on osteosarcoma of the head and neck. AB - BACKGROUND: Osteosarcoma is the most common primary bone malignancy overall but is encountered infrequently in the head and neck. Limited data are available on the significance of prognostics factors and results of therapy for patients with head and neck osteosarcoma (HNOS). It is known that surgical therapy is critical to successful outcome. The impact of adjuvant chemotherapy has not been defined well. METHODS: The National Cancer Data Base has collected data on cases of HNOS by voluntary accrual from participating hospitals. This database was used to assess patient demographics, tumor characteristics, treatment, and outcome for an 11-year period extending from 1985 to 1996. RESULTS: The data set contained 496 cases of HNOS. Overall, the 5-year disease-specific survival rate was 59.7%. Factors associated with a poor prognosis were age older than 60 years; nonmandibular tumor location; tumor size > 6 cm; histologic type of osteoblastic, not otherwise specified (NOS); advanced disease stage; nonsurgical initial therapy; and positive margins of resection. Increased use of multimodality therapy was noted during the later years of the study. No substantial difference in the 5-year survival rate was noted between treatment with surgery alone (74.7%) and surgery with adjuvant chemotherapy (71.3%), despite the greater prevalence of tumors with poor prognostic factors in the surgery and adjuvant chemotherapy group. CONCLUSIONS: Surgery with clear margins is an important factor in successful therapy for patients with HNOS. Improved survival has been noted within the last 2 decades compared with historic controls. Multiple negative prognostic factors have been identified and should be considered in treatment planning. The role of chemotherapy has not been defined well, but its incorporation into treatment of patients with high-risk tumors may improve survival. PMID- 14534885 TI - The clinical significance of soluble CD86 levels in patients with acute myeloid leukemia and myelodysplastic syndrome. AB - BACKGROUND: Levels of the soluble form of CD86 (sCD86) are elevated in a proportion of patients with leukemia. Although it is a potential modulator of antitumor responses, the significance of sCD86 in patients with hematologic malignancies is unknown. METHODS: The authors evaluated sCD86 levels by enzyme linked immunosorbent assay in patients with acute myeloid leukemia (AML) (n = 57 patients) and patients with myelodysplastic syndrome (MDS) (n = 40 patients) and analyzed the relation between sCD86 levels and various clinical parameters. RESULTS: Levels of sCD86 were elevated (> 2.32 ng/mL) relative to normal donors (0.22-2.32 ng/mL; n = 51 patients) in 25% of patients with AML and in 27% of patients with MDS. Patients with AML who had elevated sCD86 levels had significantly lower complete remission (CR) rates compared with patients with AML who had normal sCD86 levels. In multivariate analysis using sCD86 as a continuous variable and including the interaction of age and sCD86 as a variable, sCD86 was a significant prognostic factor (P = 0.014) independent of cytogenetics. Further analysis demonstrated that, in patients with AML age 60 years and younger, but not in patients older than 60 years, elevated sCD86 levels were associated with significantly shorter survival (P = 0.04). There was no correlation between sCD86 levels and CR rates or survival in patients with MDS. CONCLUSIONS: The presence in patients with AML of elevated levels of circulating sCD86 were associated with lower CR rates and poor survival. The prognostic significance of sCD86 was independent of cytogenetics but was modulated by age, in that it was independently significant only in younger patients. The results suggest that sCD86 may play a role in modulating immune responses associated with the progression of AML. PMID- 14534886 TI - Patterns of initial disease recurrence after resection of gallbladder carcinoma and hilar cholangiocarcinoma: implications for adjuvant therapeutic strategies. AB - BACKGROUND: Current approaches to adjuvant treatment after resection of gallbladder carcinoma (GBCA) and hilar cholangiocarcinoma (HCCA) are based on an incomplete understanding of the recurrence patterns of these diseases. Through an in-depth analysis of the sites of initial recurrence after resection of GBCA and HCCA, the current study aimed to highlight differences in the biology of these tumors and to provide further insight for adjuvant therapeutic strategies. METHODS: Patients with either GBCA or HCCA who underwent a potentially curative resection were identified prospectively from a maintained database. Specific sites of initial disease recurrence were identified retrospectively and categorized as locoregional (resection margin, porta hepatis, or retroperitoneal lymph nodes) or distant (peritoneal, extraabdominal, or discontiguous liver metastases). Differences in disease recurrence patterns, time to disease recurrence, and overall and site-specific survival were analyzed. RESULTS: Between May 1990 and August 2001, 177 patients underwent potentially curative resection, 97 for GBCA and 80 for HCCA. Disease recurrence and follow-up data were available for 156 patients (80 with GBCA and 76 with HCCA). The median time to disease recurrence was shorter for patients with GBCA compared with patients with HCCA (11.5 vs. 20.3 months; P = 0.007). Overall, 52 (68%) patients with HCCA and 53 (66%) patients with GBCA had disease recurrene at a median follow-up of 24 months. Of those who developed disease recurrence, isolated locoregional disease as the first site of failure occurred in 15% of patients with GBCA compared with 59% of patients with HCCA (P < 0.001). By contrast, an initial GBCA recurrence involving a distant site, with or without concomitant locoregional recurrence, occurred in 85% of patients compared with 41% of patients with HCCA (P < 0.001). This pattern of disease recurrence was diagnosis specific and did not change significantly when patients were stratified by several clinicopathologic factors, including disease stage and its component variables. Using multivariate analysis, diagnosis was an independent predictor of the site of disease recurrence. Among patients who experienced disease recurrence, survival was greater among the patients with HCCA compared with patients with GBCA (29 months vs. 20.6 months, respectively; P = 0.037). For both tumors, the site of initial disease recurrence had no apparent impact on survival time. CONCLUSIONS: After resection, recurrent GBCA is much more likely than recurrent HCCA to involve a distant site. GBCA is also associated with a much shorter time to recurrence and a shorter survival period after recurrence. The results demonstrated significant differences in the clinical behavior of these tumors and suggested that an adjuvant therapeutic strategy targeting locoregional disease, such as radiotherapy, is unlikely to have a significant impact in the overall management of GBCA. Conversely, there is at least some rationale for such an approach in patients with HCCA based on the pattern of initial recurrence. PMID- 14534887 TI - XRCC3 genetic polymorphism, smoking, and lung carcinoma risk in minority populations. AB - BACKGROUND: The XRCC3 protein participates in DNA double-strand breaks and recombinational repair. A single C-to-T nucleotide change at codon 241 (Thr241Met) has been identified in the XRCC3 gene. Using a hospital-based case control approach, the authors studied the XRCC3 polymorphism as a possible genetic risk factor for lung carcinoma in African Americans and Mexican Americans. METHODS: A total of 112 patients with lung carcinoma were frequency matched with 190 control participants on the basis of age, gender, and ethnicity. Polymerase chain reaction-restriction fragment length polymorphism molecular analysis was used successfully to identify the XRCC3 polymorphism in peripheral blood lymphocytes. RESULTS: No significant association between the XRCC3 variant allele polymorphism (odds ratio [OR], 1.25; 95% confidence interval [95% CI], 0.72-2.15) and lung carcinoma risk was noted. However, a significantly increased risk for lung carcinoma (OR, 5.20; 95% CI, 1.59-17.03) was evident in heavy smokers with the variant T-allele genotypes. Furthermore, a joint effect of the T allele and heavy smoking was observed (OR, 37.31; 95% CI, 11.43-121.72). CONCLUSIONS: The XRCC3 polymorphism appeared to be associated with increased risk of lung carcinoma in certain subgroups. It is plausible that the association is very heavily dependent on the degree of smoking. Further studies with larger sample sizes will be necessary to confirm these findings. PMID- 14534888 TI - Gemcitabine, paclitaxel, and cisplatin as induction chemotherapy for patients with biopsy-proven Stage IIIA(N2) nonsmall cell lung carcinoma: a Phase II multicenter study. AB - BACKGROUND: The objective of the current study was to define the activity and tolerability, as well as the influence on resectability, of the combination of gemcitabine, paclitaxel, and cisplatin (GTP) as induction chemotherapy for patients with Stage IIIA(N2) nonsmall cell lung carcinoma (NSCLC). METHODS: Forty nine chemotherapy-naive patients (median age, 61 years; World Health Organization performance status, 0-1) with biopsy-proven Stage IIIA(N2) disease received 1000 mg/m(2) gemcitabine, 125 mg/m(2) paclitaxel, and 50 mg/m(2) cisplatin on Days 1 and 8 of every 3 weeks until reevaluation for surgery or definitive radiotherapy. RESULTS: Grade 3-4 neutropenia was the most common hematologic toxicity, occurring in 32.7% of patients; however, only 1 case of febrile neutropenia was reported. Grade 3-4 thrombocytopenia occurred in 12.2% of patients but was not associated with bleeding. Severe nonhematologic toxicities were uncommon; the only Grade 4 nonhematologic toxicity was diarrhea, which occurred in 4% of patients. One patient died after the first course of therapy, but this event was found to be unrelated to treatment. Thirty-six patients (73.5%) achieved an objective response, and an additional 4 patients had stable disease with clearance of mediastinal lymph nodes. Overall, 29 patients underwent thoracotomy and 27 (55%) underwent complete resection. Mediastinal nodes were free of tumor in 35% of all cases, and 8 pathologic complete responses (16%) were reported. Median survival was 23 months, with a 1-year survival rate of 85%. CONCLUSIONS: GTP is highly active as an induction chemotherapy regimen for Stage IIIA(N2) NSCLC and yields good toxicity results. The use of GTP in combination with radiotherapy and new biologic drugs should be explored. PMID- 14534889 TI - Cutaneous angiosarcoma of the scalp: a multidisciplinary approach. AB - BACKGROUND: Angiosarcoma is a malignant tumor of vascular endothelial cells that arises in the head and neck. It is a rare, difficult to treat, and lethal tumor. METHODS: Clinical data from patients who were diagnosed with angiosarcoma of the scalp between 1975 and 2002 at the University of Michigan were reviewed. Analysis was performed to assess for factors impacting time to recurrence and survival. RESULTS: The study was comprised of 29 patients with a median age of 71.0 years. Most patients presented after a delay in diagnosis with either a bruise-like macule (48.3%) or a nonbruise-like nodule (51.7%). Seventy-five percent of patients had pathologic Stage T2 disease, and 76% of patients had high-grade tumors. Virtually all patients underwent surgical excision (96.6%); however, negative surgical margins were achieved in only 21.4% of patients. Multiple lesions on presentation were associated with a shorter time to recurrence (P = 0.02). The median actuarial survival was 28.4 months. Younger patients and patients with Stage T1 disease had improved survival (P = 0.024 and P = 0.013, respectively). Radiation therapy was associated significantly with a decreased chance of death (hazard ratio, 0.16; P = 0.006). CONCLUSIONS: Although surgery remains the first option for the treatment of patients with angiosarcoma of the scalp, achieving negative margins often is impossible. Patients who are younger and who have less extensive disease fare better. Postoperative radiation therapy should be employed routinely, as it may lead to improved survival. PMID- 14534890 TI - Prostate carcinoma risk subsequent to diagnosis of benign prostatic hyperplasia: a population-based cohort study in Sweden. AB - BACKGROUND: Pathologically, benign prostatic hyperplasia (BPH) is not considered a precursor for prostate carcinoma. However, because the two conditions share not only a similar hormonal environment within the prostate but also several common risk factors, it is possible that men with BPH may be at increased risk of prostate carcinoma due to these shared factors. METHODS: To elucidate this further, the authors used Swedish nationwide population-based record-linkage data to assess prostate carcinoma risk up to 26 years after the diagnosis of BPH among 86,626 men. RESULTS: Overall, relative to the general population, patients with BPH experienced little, if any, excess risk of prostate carcinoma (2% excess incidence after 10 years of follow-up). However, patients with BPH with and without surgical intervention experienced different prostate carcinoma risk patterns. Those undergoing transvesicular adenomectomy had a significant 22% lower incidence and a 23% lower mortality after the first 5 years of follow-up and those undergoing transurethral resection had a significant 10% higher incidence but a 17% lower mortality. In contrast, after the first 5 years, patients with BPH who did not receive surgical intervention experienced significant excesses of both prostate carcinoma incidence (18%) and mortality (77%). CONCLUSIONS: The differences in prostate carcinoma incidence and mortality by BPH treatment type suggest that factors related to treatment or health reasons underlying the selection of treatment influence subsequent prostate carcinoma risk. Further studies are needed to confirm the minimal excess risk of prostate carcinoma among BPH patients overall and the possible impact of BPH treatment methods on subsequent prostate carcinoma risk. PMID- 14534891 TI - Long-term efficacy and safety of zoledronic acid compared with pamidronate disodium in the treatment of skeletal complications in patients with advanced multiple myeloma or breast carcinoma: a randomized, double-blind, multicenter, comparative trial. AB - BACKGROUND: The goal of the current study was to compare the long-term (25-month) safety and efficacy of zoledronic acid with pamidronate in patients with bone lesions secondary to advanced breast carcinoma or multiple myeloma. METHODS: Patients (n = 1648) were randomized to receive 4 mg or 8 mg (reduced to 4 mg) zoledronic acid as a 15-minute infusion or to receive 90 mg pamidronate as a 2 hour infusion every 3-4 weeks for 24 months. The primary endpoint was the proportion of patients with at least 1 skeletal-related event (SRE), defined as pathologic fracture, spinal cord compression, radiation therapy, or surgery to bone. Secondary analyses included time to first SRE, skeletal morbidity rate, and multiple-event analysis. Hypercalcemia of malignancy (HCM) was included as an SRE in some secondary analyses. RESULTS: After 25 months of follow-up, zoledronic acid reduced the overall proportion of patients with an SRE and reduced the skeletal morbidity rate similar to pamidronate. Compared with pamidronate, zoledronic acid (4 mg) reduced the overall risk of developing skeletal complications (including HCM) by an additional 16% (P = 0.030). In patients with breast carcinoma, zoledronic acid (4 mg) was significantly more effective than pamidronate, reducing the risk of SREs by an additional 20% (P = 0.025) compared with pamidronate and by an additional 30% in patients receiving hormonal therapy (P = 0.009). Zoledronic acid (4 mg) and pamidronate were tolerated equally well. The most common adverse events included bone pain, nausea, and fatigue. CONCLUSIONS: Long-term follow-up data confirm that zoledronic acid was more effective than pamidronate in reducing the risk of skeletal complications in patients with bone metastases from breast carcinoma and was of similar efficacy in patients with multiple myeloma. PMID- 14534892 TI - Is the long-term survival of patients with intracranial glioblastoma multiforme overstated? AB - BACKGROUND: The 5-year survival rate for intracranial glioblastoma multiforme (GBM) has remained at 4-5% for the last 30 years, in spite of multiple randomized prospective trials. The authors hypothesized, based on the literature, that even this remarkably poor survival rate is an overstatement. They investigated this hypothesis using the the Duke University Medical Center Tumor Registry. METHODS: The authors reviewed all patients with the diagnosis of intracranial GBM recorded in the Duke University Medical Center Tumor Registry from the registry's inception in 1976 through 1996. This search identified a population of patients with a minimum of 5 years of follow-up. Each of the long-term survivors was assigned a code number for clinical information. The pathology slides were provided to a neuropathologist in a coded fashion so that the patients could not be identified. The neuropathologist reviewed the slides to analyze the presence or absence of nine histologic factors. A match technique was used to identify a control population of patients with GBM who were not 5-year survivors and were all deceased. The control population was compared with the study population to ascertain if there are histologic correlates associated with long-term survivorship. RESULTS: The authors identified 766 patients recorded by the tumor registry as having an intracranial GBM with a minimum of 5 years of follow up. Of the total population, 32 patients initially appeared to be 5-year survivors (4%). Upon review of the medical records for these 32 patients, however, the authors found only 17 patients who were truly 5-year survivors. The most common reason for miscoding was the presence of a low-grade astrocytoma that subsequently dedifferentiated into GBM. The 17 long-term survivors included 11 males and 6 females. Their mean age at diagnosis was 40.2 years. Therapy consisted of a macroscopic total resection in 4 patients (22%), a biopsy in 1 patient (6%), a subtotal resection in 10 patients (56%), and unknown extent of resection in 2 patients (11%). All patients received partial brain irradiation (mean dose, 62.6 Gy) and chemotherapy. Thirteen different single-agent or combination chemotherapy programs were used. Two patients also received I-131 monoclonal antibody therapy. Analysis of the nine histopathologic factors studied showed that intermediate fibrillary elements were more common and small anaplastic elements were less common in the long-term survivors than in the control population. CONCLUSIONS: Survival data on intracranial GBM, based on tumor registry data, should be interpreted cautiously. Reliable conclusions can only be drawn when such data are supplemented with clinical information and the histopathology is reviewed carefully. The group of long-term survivors in the current study were younger than the typical GBM population. Conventionally treated patients with GBM, chosen from an unselected population from a tumor registry, have a smaller chance of long-term survival than is generally believed. PMID- 14534893 TI - Social networks and quality of life among female long-term colorectal cancer survivors. AB - BACKGROUND: The population of long-term colorectal cancer survivors in the United States continues to increase, but little is known about how they fare-physically, mentally, or socially-in the years after diagnosis. The current study examines female long-term colorectal cancer survivors' health-related quality of life (HRQoL) in relation to social networks. METHODS: A population-based sample of female colorectal cancer survivors (n = 726) residing in Wisconsin was recontacted approximately 9 years after the initial diagnosis. Of 443 women who were alive in 1999, 307 women completed a follow-up questionnaire. Analysis was conducted on 259 participants who completed the Medical Outcomes Study Short Form 36 Health Status Survey and a modified version of Berkman and Syme's Social Network Index. Using multivariate analyses, HRQoL summary scores were tested for associations with individual and composite measures of social networks, including marital/partner status; number of children, relatives, and friends; and the frequency of religious and community participation. RESULTS: After adjusting for age, extent of disease at diagnosis, number of comorbidities, body mass, and education, HRQoL was similar to norms published for the general population. Individual social network measures (including the number of relatives and friends) and composite network measures (including network size, the number of ties seen at least once per month, and overall social connectedness) were associated positively with mental health. CONCLUSIONS: Social networks may have an important relation with HRQoL-particularly mental health-among female long term colorectal cancer survivors. The results of this study should be of interest to those seeking to understand or improve HRQoL among this growing population. PMID- 14534894 TI - Reirradiation and hyperthermia in rectal carcinoma: a retrospective study on palliative effect. AB - BACKGROUND: The objective of this study was to evaluate the palliative effect of reirradiation and hyperthermia in patients with unresectable, recurrent colorectal carcinoma. METHODS: The medical records of 54 patients with unresectable, recurrent colorectal carcinoma that caused pain and who were treated with reirradiation and hyperthermia, were evaluated retrospectively. Previous radiotherapy was given up to a total dose ranging from 25-70 grays (Gy). The median interval between prior radiotherapy and reirradiation was 22 months (range, 4-97 months). The total reirradiation dose varied from 24 Gy to 32 Gy given in fractions of 4 Gy twice weekly. Three or four hyperthermia treatments were given once weekly. Toxicity was registered. The influence of World Health Organization (WHO) performance status, maximum tumor dimension, and time between first radiotherapy and reirradiation on therapeutic outcome was evaluated. The results of this study were compared with published results on patients who received radiotherapy with or without hyperthermia. RESULTS: Forty-seven patients (87%) completed the planned treatment schedule. The maximum toxicity was Grade 2. All patients were evaluated for palliative effect. The median follow-up was 10 months. A good or complete palliative effect was achieved in 72% of patients for a median duration of 6 months. Patients who had a better WHO performance status, smaller tumors, and a longer interval between first radiotherapy and reirradiation had slightly better outcomes, although none of those parameters reached statistical significance. CONCLUSIONS: The described combined treatment was feasible and well tolerated. Comparison of results from radiotherapy plus hyperthermia with results after radiotherapy alone suggested that additional hyperthermia prolonged the duration of palliation. Firm proof of the contribution of hyperthermia will require performing a Phase III study. PMID- 14534895 TI - A multicenter Phase I safety study of intratumoral photoactivation of talaporfin sodium in patients with refractory solid tumors. AB - BACKGROUND: Photodynamic therapy (PDT) currently is approved for the palliative treatment of malignancies of the aerodigestive tract using laser-activated porfimer sodium. A new approach has been developed, based on intratumoral placement of a nonlaser light device that activates talaporfin sodium, that may expand the use of PDT to include a broader range of treatment-resistant malignancies. The safety of this approach was assessed in a Phase I study in patients with locally advanced, refractory tumors. METHODS: Twenty-one patients with radiation-resistant or chemotherapy-resistant or inoperable malignancies were enrolled in four cohorts representing four light doses. Patients were treated with a single intratumoral light device and a fixed photosensitizer dose. Safety assessments were based on review of adverse events (AEs) and serious adverse events (SAEs), and independent evaluation of computed tomography (CT) images. RESULTS: The observed occurrence of treatment-related AEs and SAEs was minimal. No cutaneous phototoxicity was observed in any patient. The overall observed tumor response rate was 33%. CONCLUSIONS: Photoactivation of talaporfin sodium using intratumoral nonlaser light was found to be safe in the patient population of the current study at all light dose levels tested. PMID- 14534896 TI - The outcome of endometrial carcinoma surveillance by ultrasound scan in women at risk of hereditary nonpolyposis colorectal carcinoma and familial colorectal carcinoma. PMID- 14534898 TI - Microsatellite instability and expression of MLH1 and MSH2 in carcinomas of the small intestine. PMID- 14534900 TI - Prediction of initial cytogenetic response for subsequent major and complete cytogenetic response to imatinib mesylate therapy in patients with Philadelphia chromosome-positive chronic myelogenous leukemia. PMID- 14534902 TI - Introduction. PMID- 14534903 TI - Siliceous spicules and skeleton frameworks in sponges: origin, diversity, ultrastructural patterns, and biological functions. AB - Silica deposition is a fundamental process in sponges. Most sponges in the Classes Demospongiae and Hexactinellida secrete siliceous elements, which can subsequently fuse, interlock with each other, or form three-dimensional structures connected by spongin. The resulting skeletal frameworks allow sponges to grow upwards and facilitate water exchange with minimal metabolic cost. Several studies on sponge skeletogenesis have been published. We are beginning to understand the mechanisms of spicule secretion and the role of spicules and skeletal frameworks in the biology, ecology, and evolution of sponges. Molecular techniques and ecological experiments have demonstrated the genetic control of the process and the contribution of environmental factors to the expression of a sponge spicule, respectively. However, other classic topics such as the role of membranes in silicon transport or whether spicules are formed in situ or secreted anywhere in the sponge mesohyl and then transported to the skeletal framework require further investigation. We review the process of silica deposition in sponges at the molecular and cellular levels, as well as the biological and ecological functions of spicules and skeletons. The genetic control of spicule shapes makes them useful in the reconstruction of sponge phylogeny, although recent experiments have demonstrated the influence of environmental factors in modulating spicule size, shape, and the presence or absence of one or more spicule types. The implications of such variations in sponge taxonomy may be important. Besides supporting sponge cells, spicules can help larvae stay buoyant while in the plankton or reach the bottom at settlement, enhance reproduction success, or catch prey. Conversely, the role of spicules and skeletons in deterring predation has not been demonstrated. Knowledge of several aspects is still based on a single or a few species and extrapolations should be made only with caution. With the advent of new molecular techniques, new lines of research are presently open and active in this field. PMID- 14534904 TI - Comparative study of spiculogenesis in demosponge and hexactinellid larvae. AB - Spicule deposition was studied by electron microscopy in fixed embryos and larvae of the haplosclerid sponge Reniera sp. and the hexactinellid Oopsacas minuta. Spicules form in centrally located vacuoles within cells and within syncytia, as in the adult sponges. In Reniera, scleroblasts differentiate from micromeres prior to gastrulation. At gastrulation the scleroblasts migrate to the periphery of the embryo and commence spicule deposition around a hexagonal axial filament. Sclerocytes have numerous pseudopodia and migrate to the posterior pole where they become aligned along the antero-posterior axis in the free-swimming larva. Reniera larvae possess some 40-50 oxeas, each 70-75 microm long and 1 microm wide. Mature Oopsacas larvae have up to 14 stauractin spicules, which are produced on a rectangular axial filament in tissues that lie under the smooth epithelium at the posterior pole of the larva. Young sclerocytes have many pseudopodia. The 4-rayed spicules elongate along both the antero-posterior and medial axes, until the longitudinal rays become anchored in a lipid-filled cytoplasm at the anterior of the larva and the lateral rays intersect around the midline. The length of the transverse rays of the stauractins in free-swimming larvae are 27-45 microm each, while the length of the two longitudinal rays is 40 80 microm. Although spicule deposition begins in cells with a similar morphology in both cellular and syncytial sponges, the elaboration and organization of the spicules differ markedly in cellular and syncytial sponges and appear to be an outcome of the very distinct cellular differentiation and larval morphogenesis that occur in each of these groups. PMID- 14534905 TI - Some aspects of silica deposition in lithistid demosponge desmas. AB - Transmitted light microscope and SEM observations of various growth stages, including very young forms, of lithistid demosponge spicules called desmas provided a wealth of new observations on silica deposition in desmas of most lithistid demosponge groups. In typical (pachastrellid) demosponges a basic feature of silica deposition in young spicules is the formation of silica granules (100-160 nm in diameter) deposited in more or less regular concentric layers. Further growth stages in typical demosponges are similar, only silica granules are smaller and more densely packed. The shape of the spicule is controlled by an organic axial filament, while features of the outer spicule surface are also determined by silicalemma. In lithistid desmas the early stage of silica deposition is controlled by an organic axial filament or, in some cases, dispersed organic molecules only. The next step, after early arrest of axial filament growth and its total encasing by silica, is the deposition of various silica granules (40-300 nm in diameter), spheres, and/or cylinders (1,300 3,330 nm), which are either the result of precipitation or the effect of templating by organic molecules (proteins and polysaccharides), without direct control by the silicalemma. The later stages of desma growth are under direct control of the silicalemma, which molds secondary branches and/or elements of sculpture of the desmas. The tips of desmas, which articulate with older desmas, are also controlled by local spatial relationships. Differentiation of morphological forms of silica in desmas, which is at least genus-specific, clearly supports the polyphyletic nature of lithistid sponges. PMID- 14534906 TI - Sponge cell reactivity to various forms of silica. AB - Several sponge species incorporate a wide range of foreign material. Whether such material is actively selected by the sponge is controversial. Here we compare the available suspended matter and the sediment incorporated in the tissue of the demosponge Chondrosia reniformis. Field observations and laboratory experiments indicate that this species selects and incorporates only siliceous materials, in particular quartz particles and opal sponge spicules, avoiding carbonate particles. The reaction of ectosome cells of Chondrosia depends on the forms of silica: after settlement of crystalline quartz particles on the sponge surface, the pinacocytes contract uniformly, giving rise to a ruffled surface that remains throughout the incorporation of foreign material. In contrast, the opal spicules elicit a motile response in pinacocytes, which cover the spicules as a result. After incorporation, while the opal spicules remain unaltered within sponge tissue, the engulfed quartz particles are quickly etched, reduced in size, and released from the sponge. The etching of quartz particles by C. reniformis is produced by ascorbic acid, and is the first evidence of such activity from the animal kingdom. Ascorbic acid has been found to change the quartz surface features, which leads to an increased radical production and a consequent dissolution of quartz. This process does not occur on opal spicules. PMID- 14534907 TI - Physical and chemical analysis of the siliceous skeletons in six sponges of two groups (demospongiae and hexactinellida). AB - The siliceous skeletons of six hexactinellids and demosponges were compared using a series of physical and chemical tests. The sponges were two hermit-crab sponges (Class Demospongiae, family Suberitidae), one from Scotland, Suberites domuncula, and the other, Pseudospongosorites suberitoides, from the Gulf of Mexico, and four hexactinellids, Hyalonema sp., Euplectella aspergillum, Rhabdocalyptus dawsoni, and Aphrocallistes vastus. The operating hypothesis was that differences in the amorphous hydrated silica skeletons in Demosponges and Hexactinellids might prove taxonomically useful. Physical properties studied included SEM, glass density, glass transition temperature (Tg), TG/DTA to determine water content, and FTIR spectra. Chemical determinations were made using energy dispersive X-ray fluorescence (EDXRF). Spicular skeletal material is deposited in concentric layers around the axial canal. With the exception of differences at several bands in the IR spectra, the siliceous skeletons in demosponges and hexactinellids are largely indistinguishable. Density was similar in all sponges (range 2.03-2.13 g/cc) and similar to the density of opal (SiO2 x 1.5H2O) (mean = 2.09 g/cc). IR spectra were similar, with prominent absorption bands at 460-470, 800, and 1,090 1,100 cm(-1) (due to different vibrational modes of Si-O-Si linkages) and at 1,650 and 3,450-3,560 cm(-1) (due to water). The skeletons of all six showed similar spectra to that of silica gel both before and after heating to 1,200 degrees C. Water comprised 10-14% of the skeleton by weight, slightly higher in the demosponges. Average spicule chemical composition was 85.2% SiO2, 12.3% water, and 2.5% other elements (mainly S, Al, K, Ca, and Na). The percent amounts of Si did not differ significantly between the demosponges and the hexactinellids. PMID- 14534908 TI - Molecular biology of demosponge axial filaments and their roles in biosilicification. AB - For hundreds of years, the skeletal elements of marine and freshwater sponges have intrigued investigators with a diverse array of remarkably complex morphologies. Early studies of demosponge monaxonal megascleres revealed the presence of a central organic axial filament running their entire length. Until recently, however, the precise function of these axial filaments was largely unknown. The spicules from the temperate Eastern Pacific demosponge, Tethya aurantia, comprise approximately 75% of the dry weight of this species, facilitating the large-scale isolation and purification of the biosilica associated proteins. Silicateins, the most abundant proteins comprising the axial filaments of these spicules, prove to be members of a well-known superfamily of proteolytic and hydrolytic enzymes and can be easily collected after silica demineralization with hydrofluoric acid. Consistent with these findings, the intact filaments are more than simple, passive templates; in vitro, they actively catalyze and spatially direct the hydrolysis and polycondensation of silicon alkoxides to yield silica at neutral pH and low temperature. Catalytic activity also is exhibited by the monomeric subunits obtained by disaggregation of the protein filaments and those produced from recombinant DNA templates cloned in bacteria. These proteins also can be used to direct the polymerization of organosilicon polymers (silicones) from the corresponding organically functionalized silicon alkoxides. Based on these observations, the silicateins are currently being used as models for the design of biomimetic agents with unique catalytic and structure-directing properties. The presence of axial filaments in a diversity of spicule types and the evolutionary implications of these findings are also discussed. PMID- 14534910 TI - Fiber diffraction study of spicules from marine sponges. AB - A synchrotron radiation fiber diffraction structural study of the axial filament of siliceous spicules from two species of marine sponges (the Demosponge Geodia cydonium and the Hexactinellid Scolymastra joubini) was carried out. The sharpness of the spots in the diffraction patterns indicated that the protein units in the filament of both samples were highly organized. A possible explanation is that the arrangement of the protein units is similar to that of the pores in highly ordered siliceous mesoporous materials. Nevertheless, the diffraction patterns are quite different for the two types of spicules. The pattern of G. cydonium is consistent with a regular 2D hexagonal lattice of protein units in the direction perpendicular to the spicule axis, with a repeating distance of 5.8 nm; the units are linked to form fibers along the axis. The pattern of S. joubini indicates the presence of two different 2D lattices in which the repeating protein units are inclined by +50 degrees and -50 degrees with respect to the elongation axis; the distance between the units increases to 8.4 nm. This 2D model is consistent with hexagonal packing of spirally oriented cylindrical protein units elongated along the filament axis. PMID- 14534909 TI - Biochemistry and cell biology of silica formation in sponges. AB - The main inorganic material forming the skeletal elements in Demospongiae as well as in Hexactinellida, the spicules, is amorphous silica. The spicules occur in the cytoplasm and the extracellular space and also in the nucleus (as silicate crystals) of some sponge cells; the function in the latter compartment is unknown. Recent evidence shows that the formation of spicules is mediated by the enzyme silicatein. The cDNA as well as the gene encoding this enzyme was cloned from Suberites domuncula. The recombinant silicatein catalyzes the synthesis of amorphous silicate using tetraethoxysilane as substrate. The enzyme is dependent on ferric iron. Silicatein also has proteolytic (cathepsin-like) activity. Incubation of primmorphs, a special form of 3D-cell aggregates, with silicon resulted in a strong increase of their size from 1-7 mm to approximately 10 mm. The morphogenetic activity of silicate is underscored by the finding that this ion increases gene expression of silicatein and collagen. Based on these findings, it is concluded that both iron and silicate stimulate the activity of silicatein. Furthermore, it is proposed that the growing spicules are surrounded by the scavenger receptor which might be considered as a docking molecule for the collagen matrix into which the spicules are embedded. PMID- 14534912 TI - Developments in the molecular biology of DYT1 dystonia. AB - The identification of a mutation of the DYT1 gene as a cause of inherited dystonia has led to many insights regarding the genetics of this disorder. In addition, there is a rapidly expanding list of inherited dystonia syndromes, the genes for some of which have been identified or localized. The DYT1 mutation has been found in a variety of ethnic groups, and it may result in a range of phenotypes. To date, studies of torsinA, the protein product of the DYT1 gene, have not revealed its function, although its widespread distribution throughout the central nervous system suggests a universal role. TorsinA has structural homology to heat shock and chaperone proteins. Evidence from studies in cell cultures and Caenorhabditis elegans, and the presence of torsinA in inclusion bodies in several neurodegenerative diseases may be indicative of a function of this nature. Preliminary studies in humans with DYT1 dystonia and in DYT1 transgenic mice suggest disruption of the dopaminergic nigrostriatal system. A functional interference with neuronal signal processing induced by mutation of torsinA is consistent with current hypotheses regarding impairment of the center surround mechanism in the striatum. PMID- 14534913 TI - Levodopa improves physical fatigue in Parkinson's disease: a double-blind, placebo-controlled, crossover study. AB - We quantitatively investigated the effect of carbidopa/levodopa (25/100) on physical fatigue during finger tapping and force generation in a double-blind, placebo-controlled crossover study. Parkinson's disease (PD) subjects were randomly assigned to carbidopa/levodopa or placebo for Visit 1 or 2 and participated in the following two studies: (1) Finger tapping. Twenty-five PD patients used their index fingers to strike two keys 20 cm apart on a musical instrument digital interface (MIDI) keyboard. The slopes of the regression line of dwell time and movement time were used to assess the rate of fatigue development. (2) Force generation. Twelve PD patients contracted the wrist extensors maximally to obtain a baseline maximum voluntary contraction (BMVC) force. Then they repetitively contracted the wrist extensors at 50% of the BMVC for 7 seconds and rested for 3 seconds. An interval maximum voluntary contraction (IMVC) was measured every three repetitions. Fatigue was defined as an IMVC of less than 60% of the BMVC. The slope of the regression line of IMVC was used to assess the rate of force decline. These two studies were repeated 1 hour after carbidopa/levodopa (25/100) or placebo. Subjects filled out the Multidimensional Fatigue Inventory (MFI) at the beginning of the first visit. Results showed that the slope of dwell time decreased with levodopa but not with placebo (P = 0.004). The rate of force decline also decreased with levodopa but not with placebo (P = 0.01). The subscores in the dimension of physical fatigue in the MFI did not correlate with the rate changes in dwell time or the rate changes in force decline. We concluded that (1) levodopa improves physical fatigue in finger tapping and force generation, (2) physical fatigue in Parkinson's disease is at least partially related to dopamine deficiency, and (3) the MFI measures different aspects of physical fatigue compared with those measured by finger tapping and force generation. PMID- 14534914 TI - Emotion recognition in patients with idiopathic Parkinson's disease. AB - Emotion recognition (ER) was examined in 64 patients with idiopathic Parkinson's disease (PD; 56 bilateral and 8 right-sided) and 64 matched healthy volunteers. Participants were administered an ER battery, consisting of the following subscores: overall ER (OER), overall facial ER, facial emotion identification (FEI) and discrimination, overall prosodic ER, and prosodic emotion identification (PEI) and discrimination. Measures of visuospatial functions, auditory attention, and depression were also administered. After controlling for visuospatial functions, auditory attention and depression, results indicated that patients with bilateral PD had poorer performance on all ER subscores, regardless of the modality and type of experimental task involved, relative to healthy volunteers. However, patients with right-sided PD had difficulty on FEI and PEI only. Whereas none of the clinical variables examined in this study predicted any of the ER subscores, visual organization and auditory attention positively predicted OER in patients with PD. In addition, visual organization also positively predicted FEI in these patients. Implications are discussed in terms of the neural substrates underlying ER. PMID- 14534915 TI - Unclassifiable parkinsonism in two European tertiary referral centres for movement disorders. AB - In view of reports on high frequencies of atypical parkinsonism from different parts of the world and in non-white communities in the United Kingdom, we have prospectively surveyed 1,000 consecutive patients with parkinsonism presenting to two European tertiary referral centres for movement disorders (London, UK, and Barcelona, Spain). The aims of our study were to assess in a cross-sectional, prospective manner the proportion of patients who could not be classified diagnostically, to identify the factors precluding classification, and to determine which diagnostic measures would increase the rate of classifiable cases. Diagnoses were established using published clinical diagnostic criteria for Parkinson's disease (PD) and for other conditions associated with parkinsonism. Twenty-nine patients in London and 25 in Barcelona were initially considered unclassifiable; nine could be classified after further investigations. Levodopa (L-dopa) responsiveness was found to have a pivotal role in establishing a clinical diagnosis in previously unclassifiable patients: In those 45 patients who remained unclassifiable, failure to respond to L-dopa without other exclusion criteria for PD was the most common finding in each centre. Our results show that 4.0 to 5.0% of parkinsonian patients presenting to specialist clinics in Western Europe cannot be categorised using currently available clinical diagnostic criteria for parkinsonian syndromes. Prolonged follow-up and neuropathological diagnosis will be needed to determine whether all these cases represent atypical presentations of established clinico-pathological entities or whether some represent unrecognised new disorders. PMID- 14534916 TI - Voxel-based morphometry detects cortical atrophy in the Parkinson variant of multiple system atrophy. AB - To determine magnetic resonance imaging (MRI) patterns of brain atrophy in parkinsonian syndromes, we applied voxel-based morphometry (VBM) to segmented gray matter, white matter, and cerebrospinal fluid compartments of T(1)-weighted brain volumes of 12 patients with probable multiple system atrophy-parkinson variant (MSA-P) and 12 Parkinson's disease patients, comparing them with 12 normal controls matched for age. In comparison to controls, a cortical atrophy pattern was observed in MSA-P patients with significant clusters of volume loss in primary sensorimotor cortices bilateral, supplementary motor areas bilateral, right premotor cortex, prefrontal cortex bilateral (middle frontal gyri) and insular cortices bilateral; subcortical atrophy occurred bilaterally in caudate nuclei and putamen as well as in the midbrain. Furthermore, an enlargement of the cerebrospinal fluid compartment was found in the lateral ventricles, third ventricle, perimesencephalic and cerebellomedullar cavities. In PD patients, significant atrophy only occurred in left caudate head with enlargement of left lateral ventricle. Comparing MSA-P to PD patients, MSA-P showed a similar cortical pattern of atrophy as compared to controls. We conclude that VBM reveals selective cortical atrophy in patients with MSA-P affecting primary and higher order motor areas as well as prefrontal and insular cortices. Further studies are required to determine clinical and/or subclinical correlates of cortical atrophy in MSA-P. PMID- 14534917 TI - Direct economic impact of Parkinson's disease: a research survey in the United Kingdom. AB - The direct costs of care were evaluated prospectively in a sample of people with Parkinson's disease (PD) in the United Kingdom in 1998. The subjects were drawn from a random sample of general practitioner practices within a representative sample of 36 Regional Health Authorities and the equivalent. A total of 444 resource use questionnaires with usable data were returned (response rate, 59%). The total mean annual cost of care per patient for all patients by age was 5,993 pounds (9,554 euro, n = 432). Hoehn and Yahr stage significantly (P < 0.001) influenced expenditure by stage as follows: 0 and I, 2,971 pounds (4,736 euro, n = 110); II, pound 3,065 (4,886 euro, n = 89); III, 6,183 pounds (9,857 euro, n = 120); IV, 10,134 pounds (euro;16,155, n = 87); V, 18,358 pounds (29,265 euro, n = 17). National Health Service costs accounted for approximately 38% and social services for 34% of the direct costs of care. Drug expenditure accounted for 24% of overall costs in the <65 years age group and 10% in patients aged >85 years. A move from home to residential care was associated with an approximately 500% cost increase. In conclusion, PD imposes significant direct costs on public services and on individuals. These costs should be taken into account when allocating public funds. PMID- 14534918 TI - Impact of placebo assignment in clinical trials of Parkinson's disease. AB - Informed consent procedures in placebo-controlled trials are developed to ensure that subjects entering studies clearly understand the possibility of placebo assignment. The extent of patient understanding and the impact of learning that they were assigned placebo treatment have not been extensively studied. By using a standardized questionnaire, we interviewed 50 consecutive placebo-treated patients from 14 placebo-controlled clinical trials of Parkinson's disease (PD) after completion of their involvement. All patients interviewed understood that their study contained a placebo arm. All had hoped to receive the study drug rather than placebo, and more than half the subjects believed they had improved clinically during their placebo exposure. Positive impressions of enrollment in placebo-controlled trials were more frequent than negative, included helping to advance science (86%); liking the experience, education, and attention associated with the clinical trial (90%); and participating in research for the benefit of other patients as well as for themselves (80%). If another placebo-controlled trial was offered, 88% expressed that they would possibly, likely, or definitely be interested in enrollment. PD patients clearly understand the concept of placebo-controlled trials they complete them, but they are inaccurate in assessing whether they received placebo treatment. Although most wish they received the active compound being tested, the overall view of participation in placebo-controlled trials is viewed very positively by PD patients. PMID- 14534919 TI - Randomized, double-blind study of pramipexole with placebo and bromocriptine in advanced Parkinson's disease. AB - We compared the efficacy and safety of pramipexole (PPX) with placebo in the treatment of advanced Parkinson's disease (PD) as an adjunct to levodopa. A bromocriptine (BR) group was included to enable determination of the noninferiority of PPX relative to BR as the standard treatment. PMID- 14534920 TI - Abnormal corticomuscular coherence is associated with the small amplitude cortical myoclonus in Parkinson's disease. AB - Coherence is the degree of time-locked correlation between two signals as a function of frequency. The purpose of this study was to test the following hypotheses: (1) corticomuscular coherence is abnormally increased in those Parkinson's disease (PD) patients with small amplitude cortical myoclonus, and (2) corticomuscular coherence peaks around the time of the myoclonus electromyographic (EMG) discharge. We studied Parkinson's disease patients with and without myoclonus and controls. The data were digitally collected and processed off-line with EMG rectification, creation of 511-msec epochs, Fast Fourier transform, and coherence analysis. In the 12 to 30 Hz frequency band, but not at 30 to 60 Hz or above, coherence peaks were observed in the PD subjects with myoclonus that were significantly greater than in the control subjects (P < 0.001) and in PD subjects without myoclonus (P < 0.001). The abnormal coherence values are evidence for abnormal rhythmic activity in cortical motor areas in those Parkinson's disease patients with myoclonus. In combination with previous findings on back-averaging, our results show that this myoclonus occurs when neuronal populations are driven to an extreme amount of synchronous activity with higher corticomuscular coherence values. These results have mechanistic implications for cortical dysfunction in Parkinson's disease and for cortical myoclonus in general. PMID- 14534921 TI - Subthalamic nucleus stimulation for Parkinson's disease preferentially improves akinesia of proximal arm movements compared to finger movements. AB - Deep brain stimulation of the subthalamic nucleus (STN-DBS) reduces akinesia in Parkinson's disease but its impact on fine motor functions was unknown. We assessed the effects of DBS and a levodopa (L-dopa) test on the timing of the precision grip in 18 patients. Improvement on UPDRS-items reflecting hand functions and the shortening of the first phases of the precision grip were more distinct in the L-dopa test than in the pure STN-DBS condition. Other akinesia items and the time for build-up of lifting force were equally improved in both conditions. This suggests that routine STN-DBS might not be equally effective on all aspects of fine motor functions. PMID- 14534923 TI - Is FDG-PET a useful tool in clinical practice for diagnosing corticobasal ganglionic degeneration? AB - Seven consecutive patients were suspected to suffer from corticobasal ganglionic degeneration (CBGD) and were studied with ([18])F-fluorodeoxyglucose (FDG) PET imaging of the brain. At the time of their FDG-PET scan, 4 of 7 patients fulfilled the clinical criteria of CBGD as proposed by Lang and associates [In Calne DB, 1994; Neurodegenerative disease. Philadelphia: Saunders]. For 2 of these 4 patients, however, an alternative clinical diagnosis was also considered. Three of the seven patients underwent an FDG-PET scan when their clinical features were not yet developed sufficiently to confirm a clinical diagnosis of CBGD. Simple visual analysis of the FDG-PET scans was carried out. All 7 patients showed an asymmetrical pattern of glucose metabolism that was demonstrated in previous studies to be characteristic in patients who had a clinical diagnosis of CBGD. The PET results helped to confirm the clinical suspicion of CBGD in 2 patients and to rule out other diagnoses in 2 other patients. For 3 patients with no sufficient symptoms to diagnose CBGD, the pattern of glucose metabolism was characteristically asymmetrical. A probable diagnosis of CBGD was made in these patients. Our results suggest that routine visual inspection of a cerebral FDG PET scan is a useful tool to confirm suspicion of the clinical diagnosis of probable CBGD, to differentiate from other hypokinetic-rigid syndromes, and to support a diagnosis CBGD in patients who do not (yet) sufficiently fulfill the clinical criteria. PMID- 14534922 TI - Is the target for thalamic deep brain stimulation the same as for thalamotomy? AB - Deep brain stimulation (DBS) has virtually replaced thalamotomy for the treatment of essential tremor. It is thought that the site for DBS is the same as the optimal lesion site; however, this match has not been investigated previously. We sought to determine whether the location of thalamic DBS matched the site at which thalamotomy would be performed. Eleven patients who had detailed microelectrode recording and stimulation for placement of DBS electrodes and subsequent successful tremor control were analysed. An experienced surgeon, blinded to outcome and final electrode position, selected the ideal thalamotomy site based on the reconstructed maps obtained intraoperatively. When the site of long-term clinically used DBS and theoretical thalamotomy location was calculated in three-dimensional space and compared for each of the x, y, and z axes in stereotactic space, there was no significant difference in the mediolateral location of DBS and theoretical lesion site. There was also no difference between the theoretical lesion site and the placement of the tip of the electrode; however, the active electrodes used for chronic stimulation were significantly more anterior (P = 0.005) and dorsal (P = 0.034) to the ideal thalamotomy target. This mismatch may reflect the compromise required between adverse and beneficial effects with chronic stimulation, but it also suggests different mechanisms of effect of DBS and thalamotomy. PMID- 14534924 TI - REM sleep behavior disorder and vocal cord paralysis in Machado-Joseph disease. AB - We evaluated the occurrence of REM sleep behaviour disorder (RBD) and vocal cord abductor paralysis (VCAP) in a group of 9 Machado-Joseph disease (MJD) patients. RBD was diagnosed by clinical history plus audiovisual polysomnography in 4 men and 1 woman (55%). While dreaming, 4 fell out of the bed and the other injured his arms. Laryngoscopy detected bilateral VCAP in 1 patient with stridor who required emergency tracheotomy, and partial vocal cord abductor restriction in 2. RBD and VCAP are two potentially injurious conditions that should be considered part of the clinical spectrum of MJD. PMID- 14534926 TI - Treatment of hypophonia with collagen vocal cord augmentation in patients with parkinsonism. AB - We conducted structured interviews of patients with Parkinson's disease (PD) and other parkinsonian disorders and their caregivers after augmentation of vocal folds with collagen injections. A total of 12 patients (mean age, 73.7 +/- 8.6 years) completed the questionnaire after an average of 3.0 +/- 1.7 collagen injections, with an average of 13.8 +/- 7.4 weeks between each injection. In addition to improvement in all six voice quality questions, there was significant improvement (P < 0.05) in the Glascow Benefit Inventory (GBI) score. The injection procedure was tolerated well by patients with an average duration of benefit lasting 7.8 +/- 8.5 weeks. Augmentation of vocal folds with collagen provides temporary improvement in voice in patients with parkinsonian hypophonia. PMID- 14534925 TI - One-year treatment with standard and sustained-release levodopa: appropriate long term treatment of restless legs syndrome? AB - To investigate the long-term efficacy and safety of sustained-release (SR) in combination with regular-release (RR) levodopa/benserazide in the treatment of restless legs syndrome (RLS), an open-label, prospective, extension study of a preceding double-blind crossover trial was performed for 12 months. Twenty-three severely disturbed RLS patients (7 men, 16 women) received a combination of RR and SR levodopa. Patients were treated on average for 10 months with a mean daily dose of 203 +/- 101 mg of RR and of 185 +/- 93 mg of SR levodopa. The mean daily total dose was 388 +/- 162 mg levodopa. Efficacy was documented using patient's rating scales, sleep diaries, and investigator's global ratings with the Clinical Global Impressions (CGI). Ten of 23 patients completed the 1-year extension. Between baseline of the crossover trial and endpoint of the extension study (last observation-carried-forward method, intention-to-treat population), quality of sleep improved (+3.5 +/- 1.9, 7-point scale), sleep latency was shortened (-131 +/- 152 minutes), and total sleep time lengthened (+ 190 +/- 136 minutes). Severity of RLS at time of falling asleep (-6.5 +/- 3.4, 11-point scale) and during the night (-6.0 +/- 3.5) was markedly lower at the end of the extension but severity of RLS during the day (+1.9 +/- 5.0) slightly increased. Of 13 dropouts, 8 patients discontinued therapy because of worsening RLS during the day. This trial shows that long-term treatment with the combination of RR and SR levodopa/benserazide in RLS patients with late-night problems was efficacious and not limited by tolerability problems in 40% of patients, whereas in the majority of patients, aggravating daytime problems required termination of the levodopa therapy within the 1-year treatment period. PMID- 14534927 TI - VIM thalamic stimulation for tremor in a patient with IgM paraproteinaemic demyelinating neuropathy. AB - We demonstrate the effect of deep brain stimulation of the ventral intermediate thalamic nucleus on intractable action tremor, in a 72-year-old man suffering from neuropathy associated with monoclonal gammopathy. PMID- 14534928 TI - Epidemiology of primary dystonias in Japan: comparison with Western countries. AB - We performed epidemiological studies of primary dystonia in the city of Kyoto. The prevalence was at least 10.1 per 100,000 persons, which was similar to that in Western countries. Facial dystonia was more common than other types, which contrasts with that reported in Europe. Age of onset for both genders was in agreement with that in other countries. PMID- 14534929 TI - Severe generalized dystonia as a presentation of a patient with aprataxin gene mutation. AB - A 14-year-old girl, homozygous for an insertion mutation of aprataxin (APTX), 689 ins T, is described. She presented with severe generalized dystonia, ataxia, ocular motor apraxia, and areflexia. The dystonia of this patient suggests involvement of the basal ganglia or thalamus, along with clinical diversity in this disorder. PMID- 14534930 TI - Periodic alternating nystagmus and rebound nystagmus in spinocerebellar ataxia type 6. AB - We report on a family with ataxia type 6 (SCA6) showing peculiar oculomotor symptoms. The proband presented with periodic alternating nystagmus (PAN), and her 2 brothers had rebound nystagmus and gaze-evoked nystagmus. They carried the identical mutation (the number of expanded CAG repeat, 24) in the CACNA1A gene. The intrafamilial variability of oculomotor symptoms may be ascribed to factors other than CAG repeat expansion size in SCA6. PMID- 14534931 TI - Brainstem myoclonus in generalised tetanus. AB - Electrophysiological studies of stimulus sensitive myoclonus in a man with generalised tetanus revealed features characteristic of hyperekplexia or brainstem myoclonus. PMID- 14534932 TI - Prevalence of Alzheimer lesions in Parkinson's disease. PMID- 14534933 TI - Quetiapine as treatment for dopaminergic-induced dyskinesias in Parkinson's disease. PMID- 14534934 TI - Persistent hemiballism after subthalamotomy: the size of the lesion matters more than the location. PMID- 14534935 TI - The 'risus sardonicus' of multiple system atrophy. PMID- 14534936 TI - Reflex sympathetic syndrome and peripheral dystonia. PMID- 14534938 TI - [Preventative biology of type 1 diabetes: implications for clinical preventative studies]. AB - Type 1 diabetes is a multifactorial disease caused by an interplay of poorly known environmental factors and partially characterized genetic factors. Focal infiltration of the endocrine pancreas by mononuclear cells and a strikingly decreased functional beta cell mass constitute the histopathological hallmarks of the disease at diagnosis, but there is a marked interindividual variability in terms of the extent of the lesions. The disease process is triggered long before clinical onset as testified by the appearance of circulating islet cell autoantibodies years before the development of hyperglycemia. After diagnosis the current insulin substitution therapy is unable to completely avoid the occurrence of the chronic complications of hyperglycemia. The design of effective prevention of hyperglycemia in subjects at risk or of a lasting cure in patients intends to eradicate the development of the invalidating chronic complications. Such clinical studies are complicated by the existence of large interindividual differences in the progression of beta cell destruction, both before and after diagnosis. Therefore it is important to first biologically characterize larger representative groups of patients and subjects at increased risk (e.g. first degree relatives), and to follow them up clinically in order to establish objective criteria for the selection of study subjects with a more homogeneous risk of beta cell destruction for participation in clinical prevention studies. In Belgium a national collaborative program--the Belgian Diabetes Registry--has allowed to collect epidemiological, clinical and biological data in more than 3,500 patients and more than 6,000 relatives. The detection of islet cell specific autoantibodies--if possible complemented with genetic and hormonal markers--facilitates the identification of subjects at high risk of rapid beta cell destruction. For example, in first degree relatives the combined presence of IA-2 antibodies and the HLA-DQ2/DQ8 genotype defines a small group of subjects (< 1%) with 75% risk of diabetes within 5 years. They qualify for participation in intervention studies aiming at beta cell mass preservation. These findings have helped to prepare several prevention studies in Belgium. PMID- 14534939 TI - [The use of fluorides in dentistry in the year 2003]. AB - During the 20th century the entire Western world has been plagued by a tremendous increase of carious lesions in the dentition of children and adults. Near the end of that century a definite decrease in the number of carious lesions was noticed. That improvement in dental health is mainly attributed to an increased presence of fluorides in the oral cavity. Fluorides can be administrated systemically or locally. Recent findings clearly suggest that a local administration is the preferred option. Correctly applicated the local use of fluorides provides a favourable and positive effect during demineralization and remineralisation processes under carious attack. Although fluorides are not the only measure in caries prevention it is widely believed that they have largely contributed to an improvement of oral health, the relief of dental pain and the safeguarding of financial means during the last twenty five years. PMID- 14534940 TI - [The relation between morphologic and functional airway changes in bronchial asthma]. AB - Asthma is currently defined as a chronic inflammatory disorder of the airways. The exact relationship between this inflammatory process and altered airway behavior in asthma remains to be fully established. More specifically, the question remains as to the exact causal relationship between airway inflammation and bronchial hyperresponsiveness (BHR), which constitutes the predominant lung function abnormality in asthma. The two main determinants of BHR are the hypersensitivity and the hyperreactivity of the airways. Hypersensitivity is reflected in a leftward shift of the dose response curve to the bronchoconstrictor effect of agonists such as histamine. More important from a clinical perspective is the hyperreactivity of the airways, which is characterized by excessive airway narrowing. The airway wall consists of three compartments, namely the inner airway wall area between the airway lumen and the smooth muscle layer, the airway smooth muscle layer and the outer airway wall area between the smooth muscle layer and the lung parenchyma. Mathematical models have calculated that changes within each of these compartments can contribute to airway hyperreactivity. Morphometric analysis of asthmatic airways confirm thickening of each of these three airway wall compartments. The contribution of airway inflammation to the thickening of each of these compartments and their relative functional impact on BHR remains to be further elucidated. Asthmatic airways display signs both of the acute and the chronic phase of an inflammatory reaction. The acute allergic inflammation is characterized by the presence of increased amounts of inflammatory cells including eosinophils, mast cells, macrophages, dendritic cells and T helper 2 (Th2) lymphocytes, and is regulated by a complex network of mutually interacting cytokines. The Th2 lymphocyte plays a crucial role within this network. Based, amongst other observations, on in vivo animal models, the hypothesis can be formulated that through the release of a range of cytokines, Th2 cells affect directly airway responsiveness. It would appear that neither crosslinking of IgE and subsequent mast cell degranulation nor eosinophil influx into the airways play a crucial role in this process. Asthmatic airways also display signs of a chronic inflammatory process, that results in more structural alterations, the so-called airway remodeling. This includes increased deposition of collagen and fibronectin, in addition to airway smooth muscle hypertrophy and hyperplasia. In vivo animal models indicate that these structural alterations have a more profound impact on BHR than the acute inflammation. These models also illustrate that depending on the exact extent and location of structural changes throughout the various airway wall compartments, remodeling can enhance but also protect against excessive airway narrowing, despite the presence of acute inflammation. These results illustrate the necessity to take into account the full extent of histological alterations throughout the airway wall, when evaluating the effect of individual cells and cytokines involved in the acute and chronic inflammatory response in asthma. PMID- 14534941 TI - [Psoriasis: an update]. AB - Psoriasis is a frequent dermatosis observed in 2 to 3% of people. It usually begin at the second decade of age but can also affect children. Diagnosis is often easily made by clinical exam. How ether many problems characterise this pathology and are not actually resolved: problems about epidemiology, etiology, treatment and psychological effects. PMID- 14534942 TI - [Myocardial perfusion scintigraphy]. AB - The aim of this review was to recall the relevance of the myocardial perfusion scintigraphy procedure in the diagnosis of coronary artery disease. Myocardial perfusion scintigraphy is undergone after treadmill exercise or pharmacological stress by infusion of Dipyridamole. Thallium-201, sestamibi-Tc99m and tetrofosmine-Tc99m are the most frequently used tracers. Gated single photon emission computed tomography provides a combined evaluation of both myocardial perfusion and function. Myocardial perfusion imaging is an accurate tool for the diagnosis of coronary artery disease with a high sensitivity 93% and good specificity 80%. Its prognostic value is now well established among patients with known coronary artery disease. In particular for risk stratification and patient management decisions. Thallium-201 SPECT is clinically accurate in assessing myocardial viability. As it is for assessment after myocardial revascularisation. Thus myocardial perfusion scintigraphy is a reliable and a cost-effective investigation for coronary artery disease. PMID- 14534943 TI - Biomedical research in developing countries: the case of Morocco in the 1990s. AB - Moroccan biomedical research occupies the third place among African or Arab countries, and its outputs considerably increased during the last decade. The quality of publications from developing countries should be improved as suggested by the comparison with developed countries. The gap between developed and developing countries is very large considering the number of publications and their quality, the number of edited journals, and the number of patented inventions, thus making developing countries more as consumers then producers. Accordingly, there is a large gap between developing and developed countries when considering the human and financial resources devoted to scientific research. PMID- 14534944 TI - [Antibiotic resistance among E. coli isolates from urinary tract infections (1999 2000): a multicenter study]. AB - E. coli is the most frequently isolate species in community as well as in nosocomial acquired urinary tract infections (UTI). Trimethoprim-sulfamethoxazole (TMP-SMX), is commonly used as empiric treatment for managing uncomplicated UTI whereas the rate of resistance to amoxicilline is increasing. A multicentric retrospective study was conducted from January 1st 1999 to December 31st 2000 to evaluate the prevalence of E. coli isolates from UTI and to guide empiric therapies. Among 6994 urinary isolates, 62.6% were resistant to amoxicilline, this rate fell to 33.6% when amoxicilline was associated to clavulanic acide, 2.7% were producing extended spectrum b-lactamases (BLSE), higher than reported in France and USA. 37.3% were resistant to TMP-SMX. The high prevalence of antimicrobial resistance among UTI E. coli isolates emphasizes the necessity to review the empiric therapies. Fluoroquinolones can be considered as an alternative therapy according to their excellent tissue penetration and their wide spectrum of activity. PMID- 14534945 TI - [A gastroenterology unit experience with adjuvant chemotherapy for non-metastatic colon cancer: results from 24 cases]. AB - A prospective study was carried from January 1996 to December 2000 including patients that received adjuvant chemotherapy type FUFOL for colon carcinoma after curative surgery. Chemotherapy was recommended for all stage Dukes B2 end C. Adjuvant chemotherapy was administered to 24 patients (13 men and 11 women, mean age 56.7 +/- 11.5 years) with surgically resected stage B 2 in 7 cases (29%), C in 17 cases (71%). Treatment was completed in 15 patients (62%). With a median follow-up of 31.6 months, 11 patients had no recurrence, one patient had locoregional recurrence and 2 patients developed liver metastasis. Adjuvant FUFOL chemotherapy is actually the recommended adjuvant post-surgical treatment for colon cancer. This valided protocol is easily realised with a low toxicity and it can be done in gastro-enterology unit. PMID- 14534947 TI - [Pressurized metered dose inhalers for the Tunisian asthmatic]. AB - For an experience done on one hundred (100) asthmatics twenty seven (27) year old, getting the "presurize doser aerosol, we fix to show how many the user of this treatment accept it as a the main goal, the inhale technique and its clinic affect. The first prescription, thirty four percent (34%) of asthma seack are afraid and judge that their disease is dangerous and not treated. The prealable learning of using technic had been realised just for seventy percent (70%) of asthmatic seattle to control the inhale technic show that only fifteen percent (15%) inhale correctly and seventy percent (70%) are not conscent of the necessity of the write coordination. To conclude, we are conducted to show and insist on the prealable learning, maintaining and finally the "auto-inhale" system which permit to contourn the problem hand-breathes (lung) cordination. PMID- 14534946 TI - Bone scan in initial staging of prostate cancer. AB - Bone scans of 64 patients with newly diagnosed prostate cancer were retrospectively analysed. Metastases were present in 29 patients (45%). In 75% of these cases, the pattern was manifeastly metastatic. The third threshold has high negative and positive predictive values. The topography of metastatic lesions is in favour of a systemic spread. There were no metastatic cases with a PSA level under 10 ng/ml. Multiple IAU and intense IAU are the most specific patterns of metastatic lesions. Also, focal lesions on sacroiliacs are also in favour of metastatic origin. The distribution of metastases is globally similar to that of the bone marrow in adult and systemic spread is the most probable. Staging bone scan must be reserved to patients with PSA level greater than 10 ng/ml, poorly degree of differentiation and advanced clinical stage. PMID- 14534948 TI - [Management of adnexal torsion]. AB - Adnexal torsion is a current emergency in gynecology because it may compromises young women fertility. We have conducted a retrospective study about 26 adnexal torsion cases managed in B service of C.M.N.T during 4 years (1997-2001). Objective of our study is to improve patient fertility prognosis; we propose a diagnosis and therapeutic attitude which respect the time between diagnosis and treatment. Laparoscopy authorizes diagnosis, treatment and prognosis of this situation. It must usually be done in young woman who declare acute pelvic pain. PMID- 14534949 TI - [Diagnosis and treatment of primary germ cell tumors of the mediastinum. Report of six cases]. AB - Primary germ cell tumours of the mediastinum are rare. We report a retrospective study of six cases. Sex ratio was 2/1. The tumor site was the posterior mediastinum in two patients and anterior in the remaining cases. High level serum markers was noted in two patients. At histologic exam we found two cases of seminoma, one embryonal carcinoma, one case of yolk sac tumor and two cases of composite tumor. All patients had non metastatic but locally advanced disease at diagnosis. Three patients were treated with cisplatin based chemotherapy followed by radiotherapy or surgical resection of residual masses. Only one patient is still alive. As it is often reported in the literature, the prognosis of this disease is poor especially for non seminomatous tumours. PMID- 14534950 TI - [Ureteritis cystica. Case report and review of the literature]. AB - The authors report one case of ureteritis cystica in 40 years old women. The diagnosis was confirmed by histologic examination. The epidemiologic, etiopathogenic, radio-diagnosis and therapeutic aspects are discussed with a review of literature. PMID- 14534951 TI - [Necrobiosis lipoidica. Report of 4 cases and review of the literature]. AB - Necrobiosis lipoidica belongs to the inflammatory granulomatous skin disorders groups with palisade reaction. The association to diabetes mellitus is classical involving 80% of diabetics in its tibial localization. A retrospective study of all cases of necrobiosis lipoidica conducted in our dermatology department over a 15-year period, found 4 diabetic patients, 3 of whom were women known as diabetic patient with extremities involvement, and 1 man not known as diabetic patient at the time of the diagnosis. Necrobiosis lipoidica appears to be a rare disorder in diabetic people and this association seems to be over estimated. PMID- 14534953 TI - [Osteoarticular manifestations of sickle cell disease]. AB - Sickle cell disease is a genetic chronic anemia caused by existence of abnormal haemoglobin. Osteo articular manifestations are often observed in the evolution of the disease, and in some cases, they inaugur the disease. Authors discuss physio-pathological mechanisms and describe main osteoarticular signs of sickle cell disease. PMID- 14534952 TI - [Congenital sick sinus syndrome in a healthy heart: case report]. AB - Isolated congenital sick sinusal syndrome on non harmed heart is a rare affection. Its association with an atrio-ventricular block is exceptional. The authors report a case of a 19 year-old patient, with an early history of bradycardia, hospitalised for effort intolerance. His electrocardiogram reveals a high degree sino-atrial block replaced by a junctional rhythm at 30/mn. During Treadmill test, the sinusal acceleration is satisfactory and an effort atrio ventricular block was present. He later had a definitive stimulation under DDDR. This report shows that the sinusal node, in the same way as the atrio-ventricular node may be injured by congenital dysimmunitary process. The coexistence of these two conductive troubles worsen the prognosis and should lead more often to the practice of definitive stimulation by the only mode DDDR. PMID- 14534954 TI - [Laparoscopic cholecystectomy. Apropos of 1570 cases]. AB - We realized a retrospective study concerning 1570 cases of celioscopic cholecystectomy screened during 6 years: from September 1995 to August 2001. There were 1328 females and 242 males with a wean age of 50 years. 256 patients underwent laparoscopic cholecystectomy for acute cholecystitis. There were one operative death. The morbidity is weak 2.1%. The rate of conversion is 12.4%. We performed 999 operative cholangiography, we found 82 common bile duct stones. Cholangiography should be systematic. PMID- 14534955 TI - Biomedical research in developing countries. I--The Algerian case (1993-1998). AB - Biomedical articles emerging from Algeria between 1993 an 1998 were analysed in order to evaluate the scientific output. By developing a search strategy based on complementary bibliographic databases, 221 articles were retrieved, which geographic location, research topic, language, journal name and impact factor were analysed. International collaborations resulted in a profound change of the publication profile in terms of language, journal impact factor, and above all a change in research interests. This study might encourage initiatives to improve literature access and contribute to define incentives for scientific research policy in Algeria as well as in other developing countries. PMID- 14534956 TI - [Acute myeloblastic leukemia in adults: evaluation of the AML 06/96 protocol]. AB - The treatment of acute myeloid leukemia (AML) permits in a population of 25 to 60 years, a complete remission (CR) about 60 to 85% with relapse free survival at 5 years from 45 to 60%. We report our therapeutic results during two years, from june 1996 to december 1998. 104 patients with the novo AML treated according to AML 06/96 protocol, the mean age was 32.5 years old, from 16 to 55 years old. The hyperleucocytar form (GB > 50,000 elts/mm3) represented the third of the cases, only 98 patients received the induction. 6 patients died before treatment. The whole rate of CR was 55%. The rate of failure was 16%, the deaths was about 15.5%, the relapse represented 30.6% with mean delay about 14.1 months, from 4 to 35 months. The CR has been maintained in 9 patients with mean recession of 53 months, from 36 months to 62 months. The overall survival at 5 years was 9%. Our results are still unsufficient in comparison with the literature and could be improved by recess of ARA-C and donorubicin doses in induction and consolidation, as well as a good knowledge about the cytogenetical aspect of the treated population. PMID- 14534957 TI - [Primary hyperaldosteronism. A study of 18 cases]. AB - Primary aldosteronism (PA) is defined as an autonomous over-production of aldosterone by the zona glomerulosa of the adrenal glands. It is classically considered as a rare cause of hypertension. Aldosterone producing adenoma (APA) and idiopatic hyperoldosteronism (IHA) represent the most frequent subtypes of PA. The authors report 18 cases of PA (14 women an 4 men) with a mean age of 42 +/- 13.5 years. All patients have hypertension and hypokaliemia (K < 3.5 mmol/l). Eleven patients have an APA, one patient has an adrenal carcinoma and three patients have IHA. The subtype of PA has not been established in three patients. We discuss in this article the clinical, biological, radiological and evolutif characteristics of our cases, and we insist on the necessity of making a precise etiologic diagnosis to propose the most adequate treatment. PMID- 14534958 TI - [Penetrating keratoplasty in children]. AB - Penetrating keratoplasty in children is a high risk transplantation. Indications increased with improvement of surgical techniques and therapies. Our purpose is to study anatomic and functional results of penetrating keratoplasty in children. We report the results of a retrospective study including 23 penetrating keratoplasties (22 eyes) performed in 20 patients younger than 16 years old, between 1991 and 2000 at the department of ophthalmology at Charles Nicolle University Hospital-Tunis. Penetrating keratoplasty was performed for congenital dystrophies in 6 cases (17.3%), for acquired traumatic opacities in 4 cases (26%) and for non traumatic acquired opacities in 13 cases (56.7%). Non traumatic acquired opacities were keratoconus in 8 cases (61.5%), keratitis in 3 cases (23%) and graft rejection in 2 cases (15.5%). The mean follow-up was 41.8 months. Grafts remained clear in 20 cases (83.3%). Graft rejection occurred in 4 cases. Graft failure occurred in 3 cases (16.6%). Mean postoperative visual acuity was 3/10 (ranging from 1/20 to 7/10). Penetrating keratoplasty in children allows a satisfying anatomic success but a moderate visual improvement. Amblyopia is the major obstacle to success in children corneal grafting. PMID- 14534959 TI - [Attitude of health personnel to organ donation and transplantation]. AB - It was shown that the rate of organ donation depends not only on the attitudes of the public but also of the predisposition of the health care staff. An opinion poll has interested 702 physicians, nurses and technicians. It comes out from this study that 59% of the sample are favorable to the organ donation. The reason of the most quoted acceptance is "humane reason" (46%). Concerning the refusal of the organ donation by the professionals of health, 3 reasons are held: (i) religious reserves (26.4%), (ii) without reasons (22.8%) and (iii) personal reasons (20.9%). For approximately 10% of the doctors the refusal is ethical. In conclusion, the Tunisian physicians adopt an attitude favorable to the organ donation, whereas nurses and technicians are rather reticent. Refusal reasons identified during this study could direct the organizers of the public awareness campaigns for the promotion of organs donation. PMID- 14534960 TI - [Hepatotoxicity of antitubercular drugs. Apropos of 28 cases]. AB - Anti-tuberculosis have up set the out come of this disease, however undesirable effects principally hepatotoxicity remain of ten the ransom of this therapy success. 28 observations of hepatic toxicity in allergology pneumo service in military hospital of Tunisia during 5 years (1996-2001) among 321 tuberculosis. Hepatic toxicity is observed in 28 cases, 21 cytolytics, 3 cholostatics and 4 mixt. 3 decease (death) have been observed. Anti-tuberculosis medicament (medecine) are sometimes this cause of many undesirable effect, which can be miner or major and even cases of decease. So that we have to take an interest at time and specialty to follow recommandation which are more and more codified. PMID- 14534961 TI - [Granuloma annulare: apropos of 18 cases]. AB - Granuloma annulare is a benign skin disorder which cause is unknown. The aim of this work is to report the results of a retrospective study dealing with 18 cases of granuloma annulare carried in the dermatology department of la Rabta hospital over a nine year period. We describe 3 clinical forms: localized granuloma annulare: 9 cases, generalized granuloma anulare: 7 cases and deep granuloma annulare: 2 cases. Diabetes mellitus was associated to granuloma annulare in 6 patients, from which 3 presented with multiple lesions. Corticosteroids were used topically in 11 cases. The outcome was marked by a partial resolution in 15 cases. In our study. We didn't found any malignancy with our granuloma annulare patients. PMID- 14534962 TI - [The overlap syndrome of autoimmune hepatitis and primary biliary cirrhosis. Clinical and therapeutic analysis of 5 cases]. AB - Autoimmune hepatitis and primary biliary cirrhosis are two different liver diseases. The aim of this study is to relate the clinical, serologic and histologic features of patients presenting the overlap syndrome, and to evaluate their response to therapy. This is a retrospective study concerning patients presenting overlap syndrome diagnosed between january 1998 and December 2001. Five patients fulfilled the diagnosis criteria of overlap syndrome. The optimal type of treatment of the overlap syndrome was not known. Treatment with UDCA or immunosuppressor alone are not efficient. Combination of these drugs should be evaluated in further studies. PMID- 14534963 TI - Neuroendocrine tumours of the digestive tract report of two cases and literature review. AB - Neuroendocrine tumors (NET) of the digestive system are rare. They comprise only 1% of all the tumors of the gastrointestinal tract. The aim of this study is to report two cases of exceptional localization of non secretary neuroendocrine tumors. Their difficulty in diagnosis, treatment and controversy in chemotherapy merit their study in depth. These are two patients that were treated in the department of general surgery (Beau Sejour) in Charles Nicolle Hospital for non secretary (NET) of the pancreas and stomach. The first was presented as a painless epigastria mass with frequent diarrhea. The second was presented as a painful mass in the left hypochondriac area without any other symptoms. Biochemical evaluation confirmed their non secretary properties. Surgery remains the first line of treatment. It was possible for the gastric, but not for the pancreatic tumor. Both were very advanced and had metastasis. Histopathological evaluation and immunohistochemical study using the method of peroxydase antiperoxydase (PAP) complexes for some special monoclonal antibodies confirmed the diagnosis. Chemotherapy of both cases was mal tolerated and was discussed in details. PMID- 14534965 TI - [A voluminous foraminal and extra foraminal disk hernia mimicking a tumor]. AB - In some cases, disc herniation can be voluminous and can then constitute a differential diagnosis with tumours. We report the case of a 46 years-old female with sciatica and crural neuralgia which resisted to medical treatment. X-rays exams were normal, but the computerized tomography showed a voluminous mass in contact with the L5 nervous root and which was developed in the psoas major muscle. The MRI revealed a voluminous foraminal and extra foraminal herniation at the L4-L5 vertebral disc, extended up to the psoas major muscle and associated with an important inflammatory infiltration. Surgical treatment was successful with a follow up of 8 months. PMID- 14534964 TI - Autologous hematopoietic stem cell transplantation for mediastinal extramedullary plasmocytoma. AB - Extramedullary plasmocytoma (EMP) is a rare cell neoplasm most frequently localised in the upper respiratory tract. We report the case of a 43 year-old man, with an unusual presentation of EMP developing in the mediastinum, two years after a diagnosis of solitary plasmocytoma of the bone which was successfully treated by local irradiation. In this aggressive presentation, we decided to perform an autologous hematopoietic stem cell transplantation. Two months after transplantation, CT scan showed disappearance of the mediastinal mass and immunofixation of the serum was normal. Selected cases of diffuse EMP, could benefit from intensive treatment followed by autologous hematopoietic stem cell transplantation. PMID- 14534966 TI - [Kikuchi disease: an unusual clinico-pathologic presentation]. AB - Histiocytic necrotizing lymphadenitis, also known as KIKUCHI disease is rare self limited condition of young adults. Manifestations include enlargements of the cervical lymph nodes, sometimes with fever, and can be associated with other non specific signs. The aetiologie of this affection is still unclear. This is a case report of a 27 year-old man who presented cervical and sus-clavicular lymphadenopathy diagnosed first as lymph nodes tuberculosis before the diagnostic of KIKUCHI disease was made. PMID- 14534967 TI - Oral biology, dental and beak disorders. Preface. PMID- 14534968 TI - Oral biology and disease in invertebrates. AB - It is hoped that this article has shown some of the amazing breadth of research into invertebrate mouthpart anatomy, development and pathology, and its relation to important disease processes that impinge on fields as diverse as environmental toxicology and antimalarial therapeutics, while also highlighting the need for further study in this field. PMID- 14534969 TI - Oral biology and conditions of amphibians. AB - This article reviews the anatomy of the skull and oral cavity in the three amphibian orders. Feeding behavior and feeding physiology are also described. A review is given of reported oral pathology. The pathology is divided into infectious causes (bacterial and parasitic) and noninfectious causes (trauma, neoplasia, and nutritional). PMID- 14534970 TI - Oral, dental, and beak disorders of reptiles. AB - Each species of reptile has evolved unique characteristics that are associated with their oral cavity, dentition, tongue, glands, and methods of mastication. These developments have provided each with an opportunity to interact with their surroundings. In captivity, the developmental relationship is often severed by the introduction of artificial surroundings and inappropriate husbandry. These changes predispose the animals to many stresses and disorders. Disorders of the oral cavity are often a representation of what is occurring systemically. Bacterial, viral, fungal, parasitic, neoplastic, and nutritional causes of disorders of the oral cavity in reptilian species are probably underestimated and likely cause a higher incidence of morbidity and mortality than has been reported. The misinformation that clients receive regarding husbandry may directly correlate with the frequency of oral diseases seen in a clinical setting. Prevention of disease of the oral cavity is primarily through providing appropriate environmental conditions and diet. PMID- 14534971 TI - Oral biology and beak disorders of birds. AB - The beak, or bill, of the bird is a complicated structure. This article describes beak anatomy as well as disorders of the beak, including trauma and diseases. PMID- 14534972 TI - Oral biology and disorders of chiroptera, insectivores, monotremes, and marsupials. AB - Oral disease occurs in all species, from the tiny marsupial mouse to the large red kangaroo, and from the smallest mole to the largest bat. Although there has been little research done in most of the species in this article, the whole range of dental and oral diseases has been recorded, including periodontal disease, caries, and many variations in position and number of teeth. Hopefully, the interest in these species will continue and further study will enable us to understand these diseases more thoroughly. PMID- 14534973 TI - Oral disorders of exotic ungulates. AB - There are many species classified as ungulates that possess a large variety of dental formulae and dental adaptations. Despite the diversity found within the ungulate group, the list of dental abnormalities remains short. Regardless of the contributing factors, cases of aveolitis accompanied by osteomyelitis ("lumpy jaw") occur more frequently than some of the other dental problems that are encountered in these species (fractures of tusks/canines, malocclusion, eruption cysts). Commonly, treatment of "lumpy jaw" requires exodontia with additional antibiotic therapy using a drug that is appropriate for the type of infection and the species of animal. Regardless of the dental abnormality, anesthetic, diagnostic, and treatment considerations typically provide the greatest challenge to the clinician in these cases. PMID- 14534974 TI - Exotic placental carnivore dentistry. AB - Dentistry in exotic carnivores is becoming more common as their level of care is enhanced. These cases present many interesting challenges, but adaptation of currently accepted dental procedures, use of newly developed instrumentation, and educated improvisation can handle almost any situation that a practitioner might encounter. It must be kept in mind, however, that there are virtually no medications, medicaments, or drugs that are approved for treatment of the various exotic carnivores. Therefore, the treatments that are discussed in this article should all be considered as "off label" use, and be reviewed closely before being applied to patients. PMID- 14534975 TI - Oral disorders of exotic rodents. AB - Our dental knowledge of rodents is still patchy but their increasing popularity and advances in technology allow us to make good strides toward better understanding. Cutting incisors with nail clippers and treating incisor problems without examining cheek teeth is no longer acceptable. Good practice dictates that a thorough examination is performed, a diagnosis is made, and treatment is planned and executed appropriately. Dentistry is expanding at a tremendous rate; the next few years should provide plenty of opportunities to solve most of the remaining problems with dental disease in rodents. PMID- 14534976 TI - Oral biology and disorders of lagomorphs. AB - Rabbit medicine, and dentistry in particular, is still at an early stage of development. With an understanding of the underlying oral physiology it is possible to devise an appropriate treatment regime for most dental problems after the nature and extent of disease has been assessed. Although many of the dental problems that are seen in practice cannot be cured, most can be controlled or managed to allow the affected rabbit to maintain a good quality of life. The continuously growing nature of the teeth makes recurrence and progression of problems the norm, so owner education and ongoing monitoring of animals is essential. By assessing the effects, beneficial or otherwise, of out treatments and communicating this to others, we will develop our knowledge and skills. Several treatments that are suggested in this article must be considered as "experimental" because they have not been assessed in large numbers of animals. If they work for you, or more importantly, if you find unexpected complications with a treatment method (as has happened with the use of calcium hydroxide paste treatment of abscess cavities) then please publicize the fact so that others can avoid the problem. Until the message on prevention can be reliably transmitted to owners, we will continue to have oral and dental problems to manage. After confidence and experience has been gained in anesthetizing rabbits it is possible to refine one's dental skills to be able to rapidly perform a thorough examination and basic treatments. Major and complex treatments require careful consideration because they may add to the animal's problems, rather than improving the situation. The best method for learning rabbit dentistry is to routinely perform postmortem examinations following euthanasia of affected animals, and spend an hour or two practicing handling the instruments and performing procedures on a cadaver. If you are not confident in your ability or do not have the best equipment for the job, the client should be informed and offered the opportunity to be referral to a "specialist." PMID- 14534977 TI - Nonhuman primate dentistry. AB - Dental procedures in nonhuman exotic primates present many out of the ordinary problems that are related to variations of the teeth and oral cavity and serious zoonotic considerations and procedural time limitations. Most of these challenges can be met with appropriate knowledge, training, equipment, and rapidity and precision in performance. Unfortunately, alacrity generally comes from repetitive performance of procedures and entails a learning curve. PMID- 14534978 TI - Oral biology and disorders of tusked mammals. AB - Tusked mammals can be terrestrial or aquatic. Many of these magnificent animals are kept in captivity all over the world. Functions of tusks vary as much as the species in which they occur. Dental anomalies and disorders of tusks and the rest of the dentition in these mammals were discussed, with an emphasis on the elephant. The tusk anatomy, with its large, conically-shaped pulp, makes it an ideal tooth for partial pulpectomy treatment in trauma cases where the pulp is exposed. Surgical techniques for tusks have been developed and were discussed. Oral tumors occur, but are rare. PMID- 14534980 TI - [Reform of health studies, 16 propositions of the Debouzie report]. PMID- 14534990 TI - [Expanded nursing competences, towards concept clarification]. PMID- 14534981 TI - [Students' health in 2003]. PMID- 14534992 TI - [New technologies in mobile services]. PMID- 14534991 TI - [Towards nursing competence evolution]. PMID- 14534993 TI - [Use of nanocrystalline silver in cancer wounds]. PMID- 14534994 TI - [Wound trauma and pain due to dressing removal, an international perspective]. PMID- 14534995 TI - ["Atraumatic dressings", a new terminology in wound care]. PMID- 14534996 TI - [Home care, realities and perspectives]. PMID- 14534997 TI - [New home occupations, enemies or partners?]. PMID- 14534998 TI - [Caring and suffering with what we are]. PMID- 14534999 TI - [What place today for community health nurses?]. PMID- 14535000 TI - [Security, quality and cost reduction....]. PMID- 14535001 TI - [Optimizing the management of wounds at home with MediSet dressing]. PMID- 14535002 TI - [Restoring the individual to his life milieu]. PMID- 14535003 TI - [Maintaining the bond at all costs]. PMID- 14535004 TI - [Which placement for the handicapped person?]. PMID- 14535005 TI - [Dying at home, between desire and reality....]. PMID- 14535006 TI - [Transmissions, you said transmissions?]. PMID- 14535007 TI - [Best use of drugs. B. Antibiotic use. 11/Bronchopulmonary infections]. PMID- 14535008 TI - [6/10 Heroin]. PMID- 14535009 TI - [Transformation of bar gene to tetraploid of Isatis indigotica]. AB - The transgenic tetraploid of Isatis indigotica mediated by Agrobacterium tumefaciens was obtained. To transfer the plant binary expression vector pCAMBIA 3300 carrying bar gene, the Agrobacterium tumefaciens strain EHA 105 was used as engineering bacterium. The results of PCR indicated that the bar gene had been transferred into and merged with the genome of Isatis indigotica. This study will make foundation for improvement of other characters of this species with genetic engineering. PMID- 14535010 TI - [Induction and identification of polyploid of Astragalus membranaceus var. mongholicus]. AB - In this article, improved agar painting method, with semi-solid of 0.2% colchicine and 0.1% agar, was adopted to treat apical buds of Astragalus membranaceus var. mongholicus (Bge.) Hsiao seedlings. Obtained plants were proved to be tetraploids by identification of biological characteristics and chromosome numbers. PMID- 14535011 TI - [Karyotype analysis of Picris davurica chromosome]. AB - OBJECTIVE: To systematically study Picris davurica Fisch chromosome karyotype. METHODS: Sections combined with micrograph were used to analyse chromosome. RESULTS: Chromosome number of normal diploid was 2n = 10, karyotype formula was K (2n) = 10 = 2 m + 8 sm, the relative length was 2n = 10 = 2 L + 2 M2 + 4 M1 + 2 S, which belonged to "3 A" type. The total length of chromosome groups was 12.51 microns, the total length of long arms was 8.36 microns, the AS. K% was 66.83%. The total volume of chromosome was 11.71 microns3. CONCLUSION: Picris davurica chromosome number and morphological character were clear. PMID- 14535012 TI - [Identification of Kochia scoparia and its substitutes by scanning electron microscope and UV spectrum]. AB - Scanning electron microscope (SEM) and UV spectrum methods were used to identify Kochia scoparia and its substitutes, including the fruits of Chenopodium album, Chenopodium serotinum and Kochia scoparia (L.) Schrad. f. trichophila Schirz. et Thell. The results showed that kochia scoparia differed from the substitutes in utride, trichome, stoma and shape of seed under SEM. The UV spectrum of Kochia scoparia was similar to that of Kochia scoparia f. trichophila, but significantly different from that of Chenopodium album and Chenopodium serotinum. PMID- 14535013 TI - [Identification of Aaron's beard with clustering analysis by attenuation reflection-Fourier transform infrared spectrometry]. AB - OBJECTIVE: To establish a new method to discriminate Aaron's beard by Fourier transformation infrared spectrometry. METHODS: Attenuation Reflection-Fourier transformation infrared spectrometry with clustering analysis was used to the identification of Aaron's beard. RESULTS: There were obvious differences in Aaron's beard. The results are consistent with that of morphologic study. CONCLUSION: This method is rapid, simple and economical, and can be used to the quality control. PMID- 14535014 TI - [Study on anti-tumor activity of extracts from cultured cells of Taxus chinensis]. AB - OBJECTIVE: To analyze the cytotoxicity and inhibitory effect of extracts from cultured cells(from stems) of Taxus chinensis on cancer cell line SMMC-7721. METHODS: MTT assay for cell viability and flow cytometry for cell cycle analysis. RESULTS: MTT results showed that the extact by dichloromethane was more effective than that by methanol on SMMC-7721, IC50 were 0.0834 mg DCW.ml-1 and 0.8002 mg DCW.ml-1 respectively. SMMC-7721 cells in G2-M stage increased with higher concentration of dichloromethane extracts from Taxus chinensis cells and longer incubation. CONCLUSION: Extracts from cultured cells of Taxus chinensis could have cytotoxic effect on SMMC-7721 and could induce apoptosis of cancer cells. PMID- 14535015 TI - [Effects of tengcha flavonoids on scavenging oxygen free radicals and inhibiting lipid-peroxidation]. AB - OBJECTIVE: To study the effects of Tengcha flavonoids (TCF) on scavenging oxygen free radicals and inhibiting lipid-peroxidation. METHODS: The effects of TCF on scavenging superoxide anion radical induced by xanthine-xanthine oxidase, inhibiting lipid-peroxidation of heart, liver, brain tissue homogenate and mitochondria induced by Fe2+ -VitC, Fe2+ -H2O2, Fe-Cys were discussed by TBA spectrometric method. RESULTS: TCF could significantly inhibit the superoxide anion radical, IC50 is 14.7 mg/L, and the lipid peroxidation of liver homogenate and mitochondria induced by auto-oxidation and induced-oxidation. The inhibition exhibited concentration-dependent manner. CONCLUSION: It indicated that TCF could scavenge oxygen free radicals and was capable of prophylacticly antagonizing against the oxidative injury by superoxide anion. PMID- 14535016 TI - [Effect of saponin from Tribulus terrestris on hyperlipidemia]. AB - The preventive and therapeutic effects of saponin from Tribulus terrestris L. on diet-induced hyperlipidemia in mice have been studied. It showed that in preventive experiment the saponin could significantly low the levels of serum TC (P < 0.05), LDL-c (P < 0.01) and liver TC (P < 0.05), TG (P < 0.05), and increase the activities of superoxide-dismutase (SOD) in liver. The therapeutic experiment showed that the saponin could significantly reduce the contents of serum TC (P < 0.05) and liver TC (P < 0.01). PMID- 14535017 TI - [Effect of Smilax china on adjunctive arthritis mouse]. AB - OBJECTIVE: To study the therapeutic action of decoction of Smilax china L. on adjunctive arthritis(AA) mouse and its mechanism. METHODS: To observe the change of secondary inflammation, thymus and spleen weights, T cell subgroup, B cell, NK cell. RESULTS: The decoction (90, 180 g.kg-1) intragastric injection (ig) could significantly inhibit AA mouse's secondary inflammatory swelling, reduce thymus and spleen weights, decrease CD4/CD8, but had little influence on B Cell. CONCLUSIONS: The decoction has therapeutic action by regulating cell-mediated immunity, but has little effect on humoral immunity. PMID- 14535018 TI - [Study on efficacy of "bushen huoxue liquid" in male rats with osteoporosis induced by dexamethasone and its mechanism]. AB - OBJECTIVE: To study the efficacy of "Bushen Huoxue Liquid" (BSHXL) in male rats with osteoporosis induced by dexamethasone (DM) and its mechanism. METHODS: Forty six male Sprague-Dawley rats at 18 months of age were chosen. They were divided into four groups, BSHXL and DM were administered appropriately for 12 weeks. General bone mineral density (GBMD), femur bending strength (FBS), serum-BGP, serum-PTH and serum-E2 were measured. RESULTS: GBMD, FBS in BSHXL prevention group were significantly higher than those in DM group (P < 0.01), and the same with those in normal control group (P > 0.05). GBMD, FBS in BSHXL treatment group were higher than those in DM group (P < 0.05), and lower than those in normal control group. Compared with DM group, level of serum-T in BSHXL prevention group and BSHXL treatment group increased (P < 0.05) and level of serum-PTH in those two groups decreased (P < 0.05). But in comparison with DM group, level of serum E2 and serum-BGP in the above-mentioned two groups had no significant difference (P > 0.05). CONCLUSION: BSHXL could prevent and treat osteoporosis in male rats induced by DM. PMID- 14535019 TI - [Study on purification of glycyrrhizic acid with macroporous adsorption resin]. AB - OBJECTIVE: To study the adsorption and desorption behavior of macroporous adsorbption resin AB-8 for glycyrrhizic acid and its effect factors. METHODS: The optimum purification conditions were investigated by high performance liquid chromatography and the orthogonal experiments. RESULTS: The optimum conditions were as follows: the flow rate at 1 ml/min, pH at 6.4-7.4 and the solution concentration at 10 mg/ml. CONCLUSION: AB-8 had a high adsorption capacity for glycyrrizic acid and elution was easy. PMID- 14535020 TI - [Study on optimum extraction conditions of alkaloids from Pinellia ternate]. AB - The optimum extraction conditions of alkaloids from Pinellia ternate (Thunb.) Breit were studied by orthogonal test. The results showed that the highest extraction rate of the alkaloids could be obtained by smashing the material in 60 (sieve number) of fragmentation and socking the material in 2.575 mol/L ammonia water, extracting alkaloids with 18 times as much chlorolform at room temperature for 25 hours. The highest extraction rate of alkaloids was 0.0817%. PMID- 14535021 TI - [Clinical observation of the treatment of 654-2 injection and "ganxian tui huang recipe" on liver cirrhosis with intractable jaundice]. AB - OBJECTIVE: To observe the effect of 654-2 injection and "GanXian Tui Huang Recipe" on hepatic fibrosis with intractable jaundice. METHODS: 60 patients were treated for 3 months and randomly divided into control group and experiment group. The experiment group was treated with 654-2 injection and "Gan Xian Tui Huang Recipe" and the control group was treated with routine treatment. All the patients have high level of Nitrogen monoxide(NO), hyaluronic acid(HA > 400 ng/ml) and hepatic fibrosis with intractable jaundice. RESULTS: In experiment group, the serum total bilirubin(TB), globulin(GLB), HA and NO were significantly decreased to normal level; 40% of portal veins were narrowed, 50% of them were not changed; 30% of spleen were thinned, 40% of them were fixed. 1 year later, 20 patients were stable and 2 patients were recurrent in 22 patients of the experiment group; in 24 patients of the control group, 7 patients were stable, 10 patients had hypersplenia, 2 patients occurred hepatocarcinoma and 5 patients were recurrent. CONCLUSION: Effect of 654-2 injection and "Gan Xian Tui Huang Recipe" on hepatic fibrosis with intractable jaundice is significant. PMID- 14535022 TI - [Promoting life style changes in the primary health care setting]. PMID- 14535023 TI - [Are ACE inhibitors and angiotensin receptor blockers useful in the treatment of atrial fibrillation?]. PMID- 14535024 TI - [The pathogenesis of psoriasis--investigating the genetics of a multifactorial disease]. PMID- 14535025 TI - [Prevention of dementia]. PMID- 14535026 TI - [Clinical picture and treatment of osteoarthritis]. PMID- 14535027 TI - [Transvaginal hydrolaparoscopy--a new method for diagnosing genital diseases]. PMID- 14535028 TI - [Basics of the drug therapy for attention deficit hyperactivity disorders in adolescents]. PMID- 14535030 TI - [Genetic fever--Internet consultation, mutation in the envelope]. PMID- 14535029 TI - [NARP syndrome--a less known mitochondrial disease]. PMID- 14535031 TI - [Recurrent headaches in a young man]. PMID- 14535032 TI - [A hand disease]. PMID- 14535033 TI - [Headache in children]. PMID- 14535034 TI - 401(k) plan asset allocation, account balances, and loan activity in 2002. PMID- 14535035 TI - [Continuous medical education in PRAXIS and online]. PMID- 14535036 TI - [Mild traumatic brain injury and its psychiatric assessment]. AB - Mild traumatic brain injuries without circumscripted necrotic areas in brain tissue were called concussion in classic terminology. It was generally assumed, that such lesions were to heal without any residues. This notion can not be upheld completely any longer, since according to today's state of knowledge on the pathogenesis of brain trauma, chronic troubles and impairments may occur even after mild traumatic brain injuries. Besides, a number of psychosocial stress factors making it more difficult to cope with the posttraumatic disturbances and the new challenging life situation, play a marked role as to the outcome. In mild brain injuries, these environmental factors as well as a general lack of internal resources may be even more crucial than the trauma's severity by itself. PMID- 14535037 TI - [New modalities and concepts in the treatment of hemorrhoids]. AB - The surgical treatment of haemorrhoids has significantly changed by introducing new techniques in the last years. Nowadays, low grade haemorrhoids, grade II and III, are easily and painfree treatable by a minimal invasive, Doppler transducer guided ligation of the haemorrhoidal arteries. In cases of circular protruding haemorrhoids, grade III and IV; the stapled mucosectomy described by Longo is also a new effective treatment. Both procedures can be performed for an outpatient or with short hospital stay and allows patients to return to work earlier compared to conventional techniques. Additionally, due to the new techniques the treatment of haemorrhoids is less painful and has increased patients' satisfaction. Therefore, the traditional haemorrhoidectomy, the Milligan-Morgan or the Ferguson procedure, has become less common and is only performed in a few special indications. PMID- 14535038 TI - [The startle reaction in emotion research]. AB - The human startle response is modulated by affective states. The startle response is facilitated in unpleasant, aversive context, such as fear, and reduced during pleasure. This association can be explored by simple technical methods. The modulation of startle is found also in animal. It offers the opportunity to investigate human emotional processes by a biologically based, language free paradigm. PMID- 14535039 TI - [A 58-year-old patient brought by his wife to the doctor's office because of amnesia]. PMID- 14535041 TI - [Doppler myocardial tissue imaging: a new promising echocardiographic technique]. AB - Tissue Doppler Imaging (TDI) is a recent ultrasound technique allowing quantification of regional myocardial function by measurement of myocardial velocities throughout the cardiac cycle. The physical principle of TDI is based on the ability of ultrasound machines to selectively display the low velocity- high amplitude Doppler signals from the myocardial walls. Three TDI modes may be used: the pulsed wave mode, the 2D color mode and the color M-mode. Several studies have already shown that TDI is a more sensitive and specific technique than conventional Doppler echocardiography for detecting moderate myocardial alterations in different settings such as hypertrophic and dilated cardiomyopathies, ischemia and heart transplant disorders. TDI might therefore be used as an accurate technique for screening cardiomyopathies, and could also represent in the next future a useful tool for evaluating the efficacy of early cardiac therapeutic interventions. PMID- 14535040 TI - [Possibilities of minimal invasive cardiac catheter interventions in the dog]. AB - The therapeutic possibilities in veterinary cardiology have developed rapidly in the past few years. Whereas until recently cardiac intervention in dogs could only be performed by thoracotomy, new minimally invasive techniques are adopted. Procedures like balloondilatation of pulmonic stenosis, coil embolisation of patent ductus arteriosus, pacemaker implantation in symptomatic bradyarrhyhtmia, and palliative balloon pericardiotomy are becoming more and more established. These alternative interventional methods are attractive, because no postsurgical pain and no complications potentially associated with thoracotomy ensue. The knowledge of such new treatment modalities and particularly the indications for an intervention are prerequisites to apply them optimally and broadly. PMID- 14535042 TI - [Cardiomyopathy, endocarditis valvularis thromboticans and pericarditis traumatica in cows--clinical and echocardiographical findings in three cases]. AB - The clinical, echocardiographical and pathomorphological findings of cows with either cardiomyopathy, endocarditis valvularis thromboticans and pericarditis traumatica are described. Based on these examples the use of echocardiography for improved diagnosis is presented. PMID- 14535043 TI - Spontaneous vegetative endocarditis due to Enterococcus faecalis in a rottweiler puppy. AB - A 5-month-old female Rottweiler was referred because of a 5-week diarrhea and a sudden onset of a cardiac murmur auscultated by its veterinarian. Definitive diagnosis of bacterial endocarditis was based on ultrasonographic visualization of vegetative cardiac lesions and positive cultures of Enterococcus faecalis in blood and urine. Complicating findings were suppurative nephritis and renal infarction. Despite intensive supportive care, the endocarditis and clinical condition deteriorated and the dog had to be euthanized. PMID- 14535044 TI - Diagnostic utility of somatosensory and motor evoked potentials in a patient with thoracic outlet syndrome. AB - OBJECTIVE: To evaluate the utility of somatosensory and motor evoked potentials in the diagnosis of neurogenic thoracic outlet syndrome (TOS). MATERIALS AND METHODS: In a female patient with progressive weakness and sensory disturbance, ulnar nerve somatosensory evoked potentials (SSEP), and intraoperative motor evoked potential (MEP) were performed. RESULTS: The SSEPs demonstrated no significant change in latency of major peaks at rest and after abduction of the shoulder. Diminishment of amplitude was noted after dynamic position of the arm. The MEPs demonstrated no significant change in latency after dynamic position. Diminishment of amplitude was noted after the dynamic position. The amplitude returned to normal after repositioning the arm. CONCLUSIONS: Electrophysiologic study is helpful in the diagnosis of neurogenic TOS. Reduced amplitude of cortical SSEP and myogenic MEP is expected during dynamic position of the affected arm. PMID- 14535045 TI - Cortical silent period in essential tremor. AB - BACKGROUND: Pathophysiology of essential tremor (ET) is controversial, current evidence indicating its genesis through a central oscillator and modulation by a peripheral component. Since transcranial magnetic stimulation induced silent period (SP) has been attributed mainly to intracortical inhibitory mechanisms, it has been found useful in assessing central motor disturbances. AIMS AND OBJECTIVES: To assess if the cortically induced SP was different in ET patients as compared to age matched healthy controls. MATERIAL AND METHODS: Twenty-four patients of ET (mean age 45.4 +/- 14.9 years) diagnosed according to criteria of Louis et al (1998) and an equal number of healthy controls (mean age 36.2 +/- 15.7 years) were recruited in the study. Cortically induced (magnetic) SP were recorded according to the standard methods described. Student's "t" test and Wilcoxon sign rank test were used for statistical analysis. RESULTS: The mean +/- SD cortical SP in ET patients (180.75 +/- 57.11 msec) in ET patients and in controls (163.83 +/- 35.65 msec) were statistically similar (p = 0.22). CONCLUSION: Our study shows that there is no significant difference in mean cortical SPs in patients with ET as compared to controls. PMID- 14535046 TI - EMG characteristics of low back and lower limb muscles during forward bending posture. AB - The study observed forward bending posture and evaluated its effect on muscular activity. The Muscular activity was monitored through the Electromyography (EMG). The objective was to extend the evaluation method of EMG due to muscular load in maintaining the posture and to investigate the existence of muscular coordination between low back and lower limb muscles during the posture. Twelve male subjects were asked to maintain the postures of six stages in bending angles from 0 to 180 degrees. The EMG of erector spinae (ES) at L1 and L5 levels, hamstrings, and gastrocnemius were recorded and analyzed by power spectral density. The characteristics of muscular activities were observed through the EMG power spectrum and the frequency. The activity changes were normalized by the relative presentation and the standardization. The relative presentation showed the level of activity changes in each posture by taking the activity during upright standing as a reference posture, and the standardization was applied to overcome the dependence of the power spectrum and the frequency upon the high standard deviation. A t test with the paired data was also applied to compare the values of EMG of the respective stages of the posture. The results showed that the activity of ES significantly increased during bending posture with an angle up to 45 degrees, and reached silent activity at 90 degrees of flexion. On the other hand, the activity of the hamstrings and gastrocnemius were recognized even when ES activity started and reached the silent activity. The lower limb muscles were considered to be always active during all stages of the posture, and they provided a greater contribution when the ES was in the silent activity. PMID- 14535047 TI - Peripheral sensory and motor abnormalities in patients with multiple sclerosis. AB - OBJECTIVES: Peripheral nerve abnormalities are uncommon in multiple sclerosis (MS). When present, they are usually attributed to factors associated with advanced disease, such as malnutrition or cytotoxic drugs or hereditary factors. However, a combination of MS and neuropathy has been reported. The aim of this study is to evaluate the question of possible peripheral involvement in MS. METHODS: We studied sensory and motor nerve conduction velocities (NCV) and amplitudes values in some sensory and motor nerves in upper and lower extremities in 20 definite multiple sclerosis patients diagnosed according to the criteria of Poser Scale and in 15 healthy subjects. The total number of studied nerves were 91 in patients group and 69 in control group. RESULTS: The most frequent electrophysiological abnormalities noted in patients group were low amplitude of the ulnar and sural nerve and slow NCV of the tibial and sural nerves. Electrophysiological abnormalities were found in 15 of 91 nerves examined (16.5%). The neurological disability was not associated with the presence of electrophysiological abnormalities. The electrophysiological abnormalities in control group subjects were a slight slow conduction of sural nerve in 2 of 69 nerves examined (2.9%). CONCLUSIONS: Our findings indicate a high frequency of sensory-motor electrophysiological nerve abnormalities in a selected group of multiple sclerosis patients. Electromyographers must be aware of these abnormalities when performing routine electromyography examinations in these patients. PMID- 14535048 TI - High-load coordination dynamics in athletes, physiotherapists, gymnasts, musicians and patients with CNS injury. AB - High-load coordination dynamics were measured in athletes, physiotherapists, gymnasts, musicians, patients with spinal cord injury and a patient with multiple sclerosis during exercise on a special coordination dynamic therapy device to quantify improvement in the central nervous system (CNS) organization due to therapy in patients and to quantify differences in the CNS organization between healthy subjects and patients with CNS injury. The values of high-load coordination dynamics for the group of athletes were two times better than those of physiotherapists, gymnasts and musicians, but still two times poorer than the best value achieved so far in a patient with a spinal cord injury after 10 months of continuous intensive coordination dynamics therapy. Especially the physiotherapists, gymnasts and musicians had poor coordination between arms and legs for the difficult intermediate coordinations between pace and trot gait for high load. Exhaustion of the CNS and improvement of CNS functioning in the short term memory could be made visible using hysteresis-like curves for load increase and decrease. When not receiving therapy, patients with CNS injury could not turn at high loads, and showed poor coordination at lower loads only. After exercising 7,000 coordinated arm and leg movements per month, the CNS organization for high load improved in 3 healthy subjects by 36%. In patients with CNS injury, such improvements of high-load coordination dynamics took several months of intensive coordination dynamics therapy including 350,000 coordinated movements per months. The rate of learning may differ in healthy subjects and patients very approximately by a factor of 50 depending on the severity of the injury. On the other hand however, the high-load coordination between arms and legs, necessary for walking could be improved during therapy even in patients with multiple sclerosis, with the consequence that they could manage better in every day life. PMID- 14535049 TI - EMG analysis of the upper and lower fascicles of the orbicularis oris muscle in deaf individuals. AB - The aim of this study was to analyze electromyographically the upper and lower fascicles of the orbicularis oris muscle in deaf patients comparing them with clinically normal patients. These patients present several disorders in facial muscles, temporomandibular joints disorders and constantly headache in virtue of not the use of the face expression muscles. This was performed in 30 patients from both sexes with a mean age of 18.5 years, divided into 3 groups of 10 patients each: 1. deaf patients with buccal breathing and incompetent lips; 2. in deaf patients with nasal breathing and competent lips; 3. clinically normal patients, in 7 clinical conditions: at rest with and without labial contact, sucking water, blowing, pursing and reciprocal compression of the lips and the emission of the syllabi "PA". It was observed that in the deaf patients group, both fascicles of the orbiculares oris muscles presented hyperactivity in most of the analyzed movements, mainly in buccal breathing with incompetent lips, and the lower fascicle demonstrated the highest electromyographic activity. PMID- 14535050 TI - Food-borne botulism cases in Van region in eastern Turkey: importance of electromyography in the diagnosis. AB - OBJECTIVES: Food-borne botulism is an acute form of poisoning that results from ingestion of a toxin produced by Clostridium botulinum. Botulism toxin causes its major effect by blocking neuromuscular transmission in autonomic and motor nerve terminals. METHODS: In this study, we present the features of eleven cases of food-borne botulism admitted to our hospital in 2001. All of the cases were caused by home-prepared foods; green beans. In these cases, the main symptoms and signs were generalized muscular weakness, dry mouth, dysphagia, disponea and diplopia. Electrophysiological studies were performed on four patients. RESULTS: Motor conduction studies showed that compound muscle action potentials were decreased with normal latencies and conduction velocities. The needle electromyography showed signs of denervation potentials like fibrillation and positive waves in four patients. Repetitive nerve stimulation with high frequency (20 Hz) induced an increment close to 100% in the amplitudes in 2 of 4 patients. CONCLUSION: Although toxin could not be detected in the patients, the electromyographic findings supported our diagnosis. We concluded that electromyography has an important role in diagnosis of botulism, especially in the condition that serologic tests are negative or cannot be performed. PMID- 14535051 TI - Electromyographic fatigue threshold of erector spinae muscle induced by a muscular endurance test in health men. AB - PURPOSE: To identify the electromyographic fatigue threshold in the erector spinae muscle. METHODS: Eight 19 to 24-year-old male volunteers participated in this study, in which surface electrodes were used, as well as a biological signals acquisition module (Lynx) with a sampling frequency of 1000 Hz, a 1000 times gain, a 20 Hz high pass filter and a 500 Hz low pass filter. The test consisted of repeated isometric contractions of the erector spinae muscle in a 45 degrees hip flexion posture, with 30%, 40%, 50% and 60% of the maximum voluntary isometric contraction. RESULTS: A positive correlation of the RMS (root mean square) value as a function of time was found for most of the subjects with 40% (N = 6), 50% (N = 7) and 60% (N = 8) loads of the maximum voluntary isometric contraction. CONCLUSIONS: It was concluded, from this study, that the proposed protocol provides evidence, through the electromyographic signal, of the development of fatigue in the erector spinae muscle with loads of 40%, 50% and 60% of the maximum voluntary isometric contraction. The protocol also allows the electromyographic fatigue threshold and its probable applicability in the diagnosis of this phenomenon during repetitive activities to be determined. PMID- 14535052 TI - M wave and H-reflex of soleus muscle before and after electrical muscle stimulation in healthy subjects. AB - PURPOSE: This study was the preliminary research for applying the evaluation of muscle fatigue using the evoked electromyography in the field of physical therapy. In this study, we speculated that muscle fatigue was induced by electrical muscle stimulation (EMS). And we studied the M wave and amplitude ratio of H/M before and after EMS because the M wave and amplitude ratio of H/M have been often used as the parameter in the study of muscle fatigue. METHODS: Subjects were five healthy males. In this study, the intermittent EMS (30 Hz) was administered to the soleus muscle of dominant leg for 10 minutes and we analyzed the amplitude of maximal M wave and the amplitude ratio of H/M in this study. RESULTS: The amplitude of maximal M wave after EMS significantly decreased compared with that before the EMS (p < 0.05). And the results of amplitude ratio of H/M varied as follows; decrease in two subjects, increase in one subject and unchanged results in two subjects. DISCUSSION: M wave reflects the excitability of muscle membrane related to the change in force during muscle fatigue and the amplitude ratio of H/M has been considered as the index of a relative excitability of alpha motoneuron pool. From the results of this study, we considered that muscle fatigue was induced by EMS as predictability because the amplitude of maximal M wave significantly decreased after EMS. And it was considered that the excitability of spinal neural function corresponding to fatigued soleus muscle by EMS was not consist change in this study. Therefore we thought that results of amplitude ratio of H/M might be influenced by excitability of spinal neural function in subject's ordinary state. CONCLUSION: It was suggested that the muscle fatigue was induced because the amplitude of maximal M wave significantly decreased after EMS in this study. And also it was suggested the excitability of spinal neural function corresponding with fatigued soleus muscle by EMS was not consist change in this study. PMID- 14535053 TI - [Effect of the Tabby mutation on the dentition of mice]. AB - The X-linked hypohidrotic ectodermal dysplasia in man leads to dental defects and is homologous to the Tabby (Ta) mutation in mouse. We currently investigate the effects of the Ta mutation on odontogenesis. The incisor germ of Ta showed an abnormal size and shape, a change in the balance between prospective crown- and root-analogue tissues and retarded cytodifferentiation. Although the enamel organ in Ta incisors was smaller, a larger proportion of the dental papilla was covered by preameloblasts-ameloblasts. The independent development of the labial and lingual parts of the enamel organ in rodent lower incisor might reflect their heterogeneous origin, as demonstrated for the upper incisor. The mandibular cheek dentition in Ta mice exhibits large variations classified in five morphotypes, based on the tooth number, shape, size and position. In Ta embryos, the mesio distal extent of the dental epithelium was similar to that in WT, but its segmentation was altered. These morphotypes could be explained by a tentative model suggesting that 1) the positions of tooth boundaries differ in Ta and WT molars and among the Ta morphotypes; 2) the tooth patterns are determined by the distal boundary of the most mesial tooth primordium while the distal teeth take advantage of the remaining dental epithelium; 3) one tooth primordium in Ta mice might derive from adjacent parts of two primordia in WT. PMID- 14535054 TI - Local application of IGF1 on dental pulp mechanically exposed; in vivo study on rabbit. AB - IGF1 (Insulin Growth Factor, 1) was intentionally applied onto pulp tissues, aiming to provoque a dentine regeneration process through the stimulation of the dentinoblasts' potententials. 72 cavities were hence performed on rabbit molars, intentionally exposing the dental pulp. Different concentrations of IGF1 were then applied; The histo and anatomo-pathological observations showed persistent vitality of the pulp without any sign of necrosis, even 6 weeks after the IGF1 application. Dentinoblasts layers (as an indication of the regeneration activity) were counted, according to a pre-established protocol, at days 7, 14, 22, 28 and 42. The type of the applied IGF1, was carefully selected to be "Binding Protein Resistant" (IGF-BPR), so to avoid any inhibition of the IGF1 action by the endogenous binding proteins (Hochscheid and coll). The results were conclusive in indicating the IGF1 as an efficient dental pulp capping product. PMID- 14535055 TI - The secretion of prostaglandin E2 and interleukin 1-beta in women with periodontal diseases and preterm low-birth-weight. AB - Prematurity is of one of the main causes of neonatal morbidity and mortality. Clinical observations show, that periodontitis in pregnant women can be a direct risk factor for preterm labor, with a greater influence rate compared to other risk factors. The aim of the study was to asses the relationship between periodontal diseases and PLBW in the population of women from the Lower Silesian Region (Poland), and the evaluation of prostaglandin E2 (PGE2), interleukin-1 beta (IL-1 beta) levels in gingival cervicular (GCF) and blood serum in women with PLBW and women giving birth on time as well as secretion of these proinflammatory mediators in whole blood after bacterial lipopolysaccharide stimulation. The study group consisted of 84 women with PLBW (39.2% primiparous), aged 17-41 (mean 27.57). The controls were 44 women (47.7% primiparous) aged 16 38 (mean 26.36) who gave birth on time to a normal birthweight baby. PGE2 and IL 1 beta concentrations in serum and GCF were determined by means of immunoenzymatic method (EIA). In the studied population women over 28 years and exposed to medical risk factors had more frequent PLBW occurrence probability. In primiparous over 28 there is 4 times greater probability of preterm labor, and in case of the severe and generalized periodontitis presence there is 3.9 times higher possibility of PLBW compared to women with healthy periodontium. In all women with PLBW there is a significantly higher PGE2 and IL-1 beta concentration in GCF, and in primiparous also PGE2 level in blood serum, compared to controls. PMID- 14535056 TI - Study on the occurrence of microorganisms on the post-surgical maxillary prostheses with obturators and in the post-surgical cavities of maxilla. AB - The bacterial environment of the mouth cavity may be subjected to change under influence of various factors, such as surgical removal of neoplasm tumors and in consequence the wearing of post-surgical prostheses with obturators. The purpose of the paper was to study the conceivable differences in occurrence of particular types of microorganisms found on the margin of post-surgical cavities and on the prosthetic obturators. The performed microbiologic examinations revealed that more pathologic bacterial flora was found on the obturators than in the post surgical cavities. The authors conclude that the post-surgical patients should pay more attention to the very accurate hygiene of their prostheses and the mouth cavity as well. PMID- 14535057 TI - In vitro evaluation of the marginal seal of four restoration materials on deciduous molars. AB - The aim of this study was to compare the marginal microleakage of Fuji II LC (A), composite resin Z250 (B), Fuji IX GP (C), and Dyract AP (F) in class V cavities and at the Fuji II LC/Z250 (D) and Fuji IX GP/composite resin Z250 (E) interfaces of an open sandwich technique on deciduous teeth. After thermocycling the mean marginal dye penetration at the enamel junction was 21.6 microns +/- 14.2 for group A; 83.6 microns +/- 32.3 for group B; 7.5 microns +/- 7.5 for group C; 38.7 microns +/- 27.5 for group D, and 0 micron for groups E and F. Mean dye penetration at the cementum junction was 37.1 +/- 20.2 (A); 123 +/- 42.1 (B); 28.7 +/- 17.1 (C); 0 (D); 14.4 +/- 14.4 (E); and 0 (F) microns. No leakage was seen at the junction between Fuji II LC and Z250 (0 micron), whereas a mean leakage of 184 microns between Fuji IX and Z250 was measured. In enamel the best seal was obtained with Dyract AP, but with differences at the limit of significance (P = 0.07). Sealing was significantly worse with Z250 (p = 0.03 versus Fuji II LC; p = 0.006 versus Fuji IX GP; and p = 0.003 versus Dyract AP). In cementum, the comparison between the grouped data Z250-Fuji II LC versus Fuji IX GP-Dyract AP was highly significant (p < 0.001), while there was no detectable difference between Z250 and Fuji II LC. PMID- 14535058 TI - [Microbial risks--from animal farming to the food]. AB - Microorganisms by far are representing the most important cause for food safety risks. Sources of food contamination are given along the whole food chain. The most frequent causative agents of food infections, Salmonella and Campylobacter, mostly can be found in the animals herds. A real improvement of the situation can only be reached by a strong inclusion of the agricultural area in the programs for repressing the pathogens. Problems in animal health and premortal stress of the slaughter animals cause considerable bacterial translocation processes and increase the consumer risks. Only few publications exist on the influence of the animal farming systems on the development of microbial food risks. The consumer accepts only animal management systems, which meet the demands for animal welfare, economical efficiency and especially the demand for product safety. PMID- 14535059 TI - [Effects of animal housing facilities on residues and contaminants in food]. AB - There are--especially in the case of food of animal origin--only a few well controlled studies available comparing organically and conventionally produced food with a focus on residues and contaminants. The differences found seemed to be much lower than expected, and the amounts of residues and contaminants were mostly below regulatory maximum residue levels. In the case of organically and conventionally produced milk there have been slight but usually not significant differences reported. One important observation is, that conventionally produced milk contains aflatoxin M1 more frequently than biologically produced milk. The investigation of eggs from different housing systems for laying hens showed in the case of dioxins the highest concentrations in free range eggs. In contrast, propoxur, an insecticide used against the poultry red mite, was found in eggs from battery cages in higher amounts than in those from enriched cages and in eggs obtained from an aviary system. Further research in this field is highly recommended, but there should be more sophisticated evaluation of the data sets from national and international monitoring programs. PMID- 14535060 TI - [Consumer expectations for livestock farming and food safety]. AB - Food safety optimization measures neglect the consumers' psychological requirements. To avoid guilt--delight-discords, problems of livestock farming are not registered in the first place. Food checks at the purchase are felt to be a matter of the consumer's own competence, using his five senses and primarily seen as a tasting test. Market mechanisms supply additional safety in the background. Consumer behavior in acute crises must be seen from the point of protective magie. Governmental pre-tests raise doubts on the consumer's shopping competence, they are felt to be insulting and unreliable. Certificates and quality seals only stand a chance as complimentary service to the consumer's own competence. PMID- 14535061 TI - [Poultry husbandry and animal health]. AB - Close interactions are existing between poultry husbandry and poultry health. The more housing systems and the environment of the animals can be controlled, the less the general risk of disorders in poultry flocks--especially of diseases which are caused by the introduction of microoganisms. Resulting deterimental effects will affect not only the animals themselves, but also pose a risk indirectly for humans via food originating from animals under production. Also, by keeping the risk of infections as low as possible, the use of therapeutics can be avoided. This will reduce the risk of residues in food of animal origin. In summary, with all probability open poultry husbandry systems, especially those including free range systems pose increased risks for poultry health and consequently for the quality of food originating from poultry production. At least, those systems require highest standards of biosecurity, defined as management, location, farm layout, cleaning and desinfection incl. pest control programs, immunization and specific veterinary monitoring concepts to prevent infections. PMID- 14535062 TI - [Parasitological risks--from animal husbandry to food and humans]. AB - Livestock management influences the infection risk with parasites. Extensive outdoor management increases the well being of the animals, but may also increase the infection risk with vector or intermediate host transmitted parasites. Several parasites can be transmitted by food. Trichinellosis again represents an increasing threat to human health by newly discovered species, new reservoir hosts and increasing meat import from eastern Europe. Tapeworm infections with Taenia saginata are more frequent than reported in the official statistics. The accurate prevalence and ways of distribution are not known. Toxoplasma gondii is a serious threat to pregnant, non-immune women and their babies. Cat faeces and pork meat are the most important sources of infection. Information, prevention of transmission and diagnostic examination of endangered persons are necessary to minimise the infection risk. Sarcocystis suihominis forms cysts in muscles of pigs, which may cause gastrointestinal symptoms after oral ingestion. In most cases however this protozoan parasite is not or only transiently harmful. PMID- 14535063 TI - [Housing- and management-specific effects on the health of swine]. AB - Important diseases with regard to inadequate housing and management conditions in pig production are given. Typical examples like claw lesions, cannibalism and infectious factorial diseases are described. Beside negative factors of pig housing and management which are of particular pathogenic importance some aspects of disease control and herd health are listed finally. PMID- 14535064 TI - [Practical information on classical fowl plague (highly pathogenic avian influenza)]. AB - This brief review summarises some structural and biological properties of the highly pathogenic avian influenza virus and its biological significance for animal and man. Sources of actual information in case of an acute disease outbreak are also given. PMID- 14535065 TI - [Case report--polydactyly in a German holstein calf]. AB - A female German Holstein twin calf showed polydactyly in both front limbs and in one of the hind limbs. In three limbs an additional medial toe was seen. No malformations of the limbs were present in the male twin partner and four maternal half sibs. The analysis of the pedigree revealed that the affected calf had common ancestors. The coefficient of inbreeding of the affected calf was 1.312%. As a monogenic or digenic inheritance was suspected in similar cases, it may be probable that the polydactyly observed here has a genetic background. PMID- 14535066 TI - [What role for fluoroquinolones in intensive therapy?]. PMID- 14535067 TI - [What role for glycopeptides in intensive therapy?]. PMID- 14535068 TI - [What role for streptogramin in intensive therapy?]. PMID- 14535069 TI - [Evaluation of a number of commercial diets for iguana (Iguana iguana), bearded dragons (Pogona vitticeps), and land and marsh tortoises]. AB - Fifteen commercially available, complete diets for iguanas, bearded dragons, and chelonians were analysed and compared with the ideal composition of a diet formulated by the authors. The clinical risks of feeding with a too high or a too low content of specific nutrients are described. Nutrient deficiencies are not expected if these diets are the sole source of nutrients. However, one diet had only a marginally adequate protein content, five diets for herbivorous reptiles contained a too low percentage of crude fibre, three diets contained an undesirably high percentage of calcium, and three diets had an extremely high iron content. As a rule, the calcium and iron contents of diets are not declared on the food label, which complicates evaluation of the diets. PMID- 14535070 TI - [Predecessors: veterinarians from earlier times (52). James Law (1838-1921)]. PMID- 14535071 TI - [Waiting times in animal health care?]. PMID- 14535072 TI - [Hypercalcemia in the dog and cat]. PMID- 14535073 TI - [Introduction to Floor Stembert, with publication about her research employment disability. "We don't have to reinvent the wheel!" (Interview by Lisetta Verkleij)]. PMID- 14535074 TI - [Veterinarian: a healthy profession?]. AB - Data of the insurance company Movir reveals that veterinarians are more often occupationally disabled compared to other occupations. The causes for this disability are: problems with the locomotion apparatus, accidents and psychological problems. Regarding the complaints of the locomotion apparatus is found that proportionally between 51 and 65 years there are more vets disabled. In absolute numbers the majority of the problems at Movir involves the farm animal vet. About the relative importance or indemnities of other insurance companies no statements can be made. The localisation of the affections are: lower back, knees, shoulders, neck and wrist/hand. Regarding the accidents is found that a lot of problems happen in the private situation, but that the high percentage of disability caused by accidents compared with other professions is probably caused by the profession related accidents. From the numbers of Movir comes forth that especially working with horses is dangerous, but there is no specific veterinarian skill very risky. Regarding the psychological problems comes forth that there are no differences between men and women. Proportionally from 46 years until 65 years there is a higher incidence of burnout then at younger ages. In absolute numbers more companion animal vets are burnout, but no statements can be made about the relative importance. There is no difference at Movir between the solopractice and the partnership regarding burnout. The professional circumstances playing a role in developing burnout are: a too high workload, problems with colleagues within the partnership, irregular shifts, organisation and management and financial management. The personal aspects of these veterinarians that play a role in burnout are: high expectations and demands, perfectionism and insecurity about the own abilities. In many of the files there were also problems within the private situation. PMID- 14535075 TI - [Career planning....also for veterinarians?]. PMID- 14535076 TI - [Dentists easily win from veterinarians]. PMID- 14535077 TI - [Congress member Hendrik Tolsma about the Yearly Congress 2003. "Intercollegial contacts can contribute to better communication within the veterinary world." (Interview by Bert Hendriks)]. PMID- 14535078 TI - Should you outsource your transcription? PMID- 14535079 TI - CMS issues HIPAA transactions compliance guidance; questions remain. PMID- 14535080 TI - Medicare conferees reach agreement on regulatory relief provisions. Long advocated MGMA issues decided favorably. PMID- 14535081 TI - Good doctor/good leader. PMID- 14535082 TI - Comin' to the country. Recruiting physicians to rural communities. PMID- 14535083 TI - Make your last step a smart step. Post-investment audit--the last step of the capital budgeting process. PMID- 14535084 TI - Strategic planning: it's worth the effort. Homework makes retreats work. AB - Medical groups that achieve superior operational and financial performance frequently have leaders who take time to hold strategic planning retreats. These gatherings allow practice leaders to address difficult issues such as expansion, compensation structure and governance--away from the workplace and facilitated by an outsider. The result: organizational direction that all leaders help shape and commit to. PMID- 14535085 TI - Come hither. Recruiting today's disillusioned medical residents. AB - It seems that one-fourth of today's medical residents would choose another field if they had their education to do over again. Why the disillusionment with medicine? This article discusses the reasons and offers guidelines for recruiting young physicians who approach employment with uncertainty, skepticism and priorities that differ from those of the previous generation. PMID- 14535086 TI - Four-square. Practice profitability stands on four foundations. AB - A medical practice's profitability stands on four legs: physician productivity, accounts receivable, overhead costs and ancillary revenue. The author describes where weakness can occur in each of these foundations and how to remedy such structural defects. PMID- 14535087 TI - Addressing uncompensated hospital services. Case study: physician revenue enhancement program. AB - Traditionally, physicians have provided many uncompensated services to hospitals such as emergency room on-call duty, charity care and committee appointments. But physicians can no longer afford to give away services, and hospitals need their assistance to become more cost-efficient. This article describes an innovative physician-driven cost-management program that pays physicians for their hospital work and saves the hospital money. PMID- 14535088 TI - [Study of 8-OH-dG and its correlation with several cancer related gene in lung cancer tissues]. AB - To investigate the relationship among 8-OH-dG and the development of human lung cancer and cancer related genes, an 8-OH-dG-specific monoclonal antibody and biotin-streptavidin immuno-staining were used to detect the 8-OH-dG in 150 cases of human lung cancer tissues, 120 adjacent lung tissues without cancer cells, 40 benign lung lesions and 40 normal lung tissues. The expressions of P53, C-MYC, K RAS, BCL-2 and hTERT(human telomerase reverse transcriptase) were determined by immunohistochemistry and the relationship among the 8-OH-dG and these genes was analyzed. The 8-OH-dG were positive in 139 of 150 (92.7%) lung cancer specimens, and the percentage of adduct labeling cell in lung cancer specimens was (24.00 +/ 25.11)% (mean +/- SE). 21 of 120 (17.5%) adjacent lung tissues were adduct positive, and the percentage of adduct labeling cell was 2.42 +/- 5.98%. 4 of 40 (10.0%) benign lung lesions were adduct positive, and the percentage of adduct labeling cell was 0.80 +/- 1.30%, whereas 2 of 40 (5.0%) normal lung tissues were weak positive with 8-oh-dG, and the percentage of adduct labeling cell in this group was (0.34 +/- 1.01)%. The level of 8-OH-dG in lung cancer tissues was significantly higher than that of adjacent lung tissues, benign lung lesions and normal lungs (P < 0.01). The lung cancer patients were stratified by sex, age, cell types and smoking history, but these characteristics were not correlated with the level of 8-OH-dG. In the investigation of the relationship between the 8 OH-dG and five cancer related genes, higher 8-OH-dG levels were observed in lung cancer patients with over-expression of K-RAS and BCL-2 than those of negative expressed patients (P-value were 0.035 and 0.034 respectively), whereas the expression of P53, C-MYC and hTERT were not correlated with level of 8-OH-dG. 8 OH-dG was an important biomarker that may reflect the oxidative DNA damages of cells, and 8-OH-dG may affect K-RAS and BCL-2 genes in the carcinogenesis of lung cancer. PMID- 14535089 TI - [Effect of arsenic toxicity on morphology and viability of enzyme in primary culture of rat hippocampal neurons]. AB - In order to study the toxic consequences of arsenic at different levels on hippocampal neurons of rat, different doses of sodium arsenite (0, 0.08, 0.40, 2.00 and 10.00 mg/L) were given to observe the changes of viability, morphological and biochemical parameters. The results showed that the arsenite content in primarily cultured rat hippocampal neurons decreased in growth of viable cells. The morphometric analysis showed that the isolated neonatal hippocampal neurons exposed to lower concentration of arsenic had cytoplasm vacuoles, frothing and nuclear condensation with the intact membrane, indicating the possible induction of apoptosis, whereas higher dose of arsenic leaded to the appearance of necrosis. The degeneration of cytoplasm with the appearance of large vacuoles and loss of cell-cell junction were also observed. Meanwhile the Nissles decreased in the cytoplasm confirmed by the Tolidine blue. Biochemically determined increase in the viability of L-LDH showed increased in extra-cellular release as well as decrease in AchE. Some results showed the dose-time dependent relationship. It is suggested that the neonatal hippocampal neurons of rat treated with different doses of arsenic suffered from morphological changes including degeneration, apoptosis and necrosis in various degrees. PMID- 14535090 TI - [Effects of lead on somatostatin positive neuron and its mRNA expression in rat's hippocampus]. AB - In order to study the changes of hippocampal SS and SSmRNA positive neurons, Wistar rats were exposed to lead by drinking 2000 micrograms/ml lead acetate solution for three months, the blood and hippocampal lead concentration were measured with atomic absorption spectometry, and the number of hippocampal SS and SSmRNA positive neurons were observed with immunohistochemistry, and in situ hybridization respectively. The results showed that the blood and hippocampal lead content in lead-exposed rats were obviously higher than those of the control rats (130.05 +/- 14.21) mg/L and (1011.62 +/- 59.54) ng/L respectively v.s (5.54 +/- 0.76) mg/L and (181.98 +/- 47.27) ng/L respectively (P < 0.05). The number of SS and SSmRNA positive neurons in CA1 area of lead-exposed rats were significantly smaller than those of the control rats (23.27 +/- 4.58 and 37.60 +/ 9.69 respectively v.s. 40.38 +/- 7.61 and 53.48 +/- 10.22 respectively, P < 0.01). The numbers of SS and SSmRNA positive neurons in CA3 area of lead-exposed rats were smaller than that of the controls rats (12.16 +/- 3.40 and 16.18 +/- 6.19 respectively v.s 17.68 +/- 5.86 and 25.93 +/- 7.47 respectively, P < 0.05). The study suggested that the hippocampal SS and SSmRNA positive neurons of lead exposed rat have a lower expression of SS and SSmRNA. PMID- 14535091 TI - [Distribution of microcystin-LR and anatoxin-A and their influence factors in Dianshan Lake during summer and autumn]. AB - Six different sampling locations in Dianshan Lake were selected to study the distributions of algaes and algae toxins and their influencing factors in summer and autumn. Total nitrogen (TN), total phosphorous (TP), Fe, chlorophyla(Chla) and COD in water samples were determined. At the same time, water temperature (TEM), pH value, illumination, turbidity degree (TUR) of water body were recorded. In addition, the total number and attribute of algae cell and the content of microcystin-LR(MC-LR) and anatoxin-A(ANTX-A) were checked. The results showed that the pollution of both toxins was most serious in July. The MC-LR were mostly distributed in Wangxiang, but the ANTX-A were mostly distributed in Jishuigang. Generalized estimating equations (GEE) regression analysis showed that total number of algae cell was significantly correlated with TN, TP and Chla (P < 0.05). The distribution of MC-LR was significantly affected by total number of algae cells and TEM (P < 0.01), but TUR mainly affected the distributions of ANTX-A(P < 0.01). There was weak correlation between both toxins (P < 0.05). In short, there was a serious pollution of MC-LR, but ANTX-A pollution was light. The factors influencing the distributions of both algae toxins were not completely consistent. PMID- 14535092 TI - [Study on food contaminants monitoring in China during 2000-2001]. AB - The contamination levels of 10 main kinds of foodstuffs in 11 provinces and cities were monitored by the national food contamination monitoring system from 2000-2001. About 45 thousand data were obtained in strict quality control. The data indicated that the contamination level of lead was increased in 10 years in meat, eggs, milk-based products especially in preserved eggs, but the contamination levels of arsenic, cadmium and hydrargyrum were not serious by adopting the national standards. Organochlorine pesticides residues (666, DDT) maintained downtrend and organophosphorus pesticides residues met the national associated criteria except methyl parathion in cereals. PMID- 14535093 TI - [Effects of amino acids on the survival of heat-exposed Drosophila]. AB - To investigate the effects of amino acids on the survival of heat-exposed Drosophila and to select the amino acids that can protect the Drosophila from heat exposure, Drosophila melanogaster were fed with basic culture in 25 degrees C. The male virgin Drosophilas were distinguished from female. Both male and female virgin Drosophilas were divided respectively to the trail groups and control group with randomization. The trail groups were fed with culture supplying different amino acids in 37.5 degrees C. After 10 hours, the dead insects were counted every two hours until all of them were dead. The mortality rates of heat-exposed Drosophilas that feed in the culture added with the glutamine, arginine, phenylalanine, tyrosine and glutamic acid were obviously higher than that of control. The glutamine, arginine, phenylalanine, tyrosine and glutamic acid might increase survival time of heat-exposed Drosophila, but a dose response relationship was not observed. PMID- 14535094 TI - [Effects of cisplatin and sodium selenite on transformation and rRNA gene transcription activity of human T lymphocytes]. AB - In order to investigate the effect of sodium selenite on the action of cisplatin inhibiting transformation of human T lymphocytes, explore the influence of cisplatin and sodium selenite on rRNA gene transcription activity of lymphocytes and to explore the relationship between transformation of T cells and rRNA gene transcription activity, cisplatin was administered to the culture of lymphocytes after incubation for 24 hours with three different dosages (0.05, 0.2, and 0.5 mg/L). The culture was also supplemented with sodium selenite (0.05 mg/L) at the beginning of the cell culture or at the same time as cisplatin was administered for different treatment. The slide specimens were prepared using cytogenetical method after the cells were cultured for 72 hours, and then, the changes of the transformation rate of T lymphocytes and frequencies of Ag-NORs that reflect rRNA gene transcription activity were investigated. The results showed that the rate of T lymphocyte transformation in the groups treated with both sodium selenite and cisplatin were significantly increased (p < 0.01) as compared with that in the groups without sodium selenite supplementation and suffering exposures to cisplatin alone at corresponding dose, up to or over the rate of negative control group. There were no significant differences in the ratios of lymphocyte transformation between the groups pre-treated by sodium selenite and the groups treated by both sodium selenite and cisplatin on the condition that the other treatments were the same. There were no remarkable differences (p > 0.05) in the frequencies of Ag-NOR and satellite association among the all groups. This study indicated that sodium selenite supplementation can preclude the inhibition of transformation of T lymphocytes in vitro by cisplatin, regardless the pre treatment by sodium selenite or co-treatment with cisplatin. PMID- 14535095 TI - [Study on the effect of the growth and aflatoxin production by Aspergillus flavus parasiticus NRRL 2999 in the present of Lactobacillus plantarum ATCC 8014]. AB - The effect of L. plantarum ATCC 8014 on growth and aflatoxin B1 production of A. flavus subsp. parasiticus NRRL 2999 was studied and determined. A spore suspension of Aspergillus flavus parasiticus NRRL 2999 is inoculated into MRS medium containing L. plantarum and incubated at 28 degrees C for 15 days. The pH of the medium, weight of mycelia and aflatoxin B1 production were determined on day 3, 6, 9, 12, 15 of incubation. The growth of A. flavus subsp. parasiticus NRRL 2999 was inhibited when spores were added to a 24 h L. plantarum ATCC8014 culture and when both organisms were added simultaneously. The inhibition of the growth of A. flavus subsp. parasiticus NRRL 2999 was probably due to inactivating the viability of spores. The spores of A. flavus subsp. parasiticus NRRL 2999 became swollen in the presence of L. plantarum ATCC8014 culture, at the same time, and A. flavus subsp. parasiticus NRRL 2999 also affected the shape of L. plantarum ATCC8014 cells, which became enlarged. However, when L. plantarum ATCC8014 was added to a 3-day A. flavus subsp. parasiticus NRRL 2999 culture, there was not effect on mold growth and aflatoxin B1 production. Lactic acid inhibited growth of A. flavus subsp. culture in the MRS medium with initial pH value of 4.5. However, the production of aflatoxin B1 was not affected. PMID- 14535096 TI - [Investigation of concentration of microcystin in raw materials and finished products of spires health food]. AB - In order to investigate the concentration of micromystin in raw materials and finished products of spires health food, we collected 33 water samples from water resources, 160 water samples from culturing pool, 86 samples of Spires slurry and 70 samples of powders of Spires raw materials from 7 Spires manufacturers in Jiangsu, Yunnan, Fujian and Guandong provinces which are the main spires producing areas from July to August 2002. At the same time, we also collected 71 samples of 19 finished products of Spires health foods in markets of above mentioned 4 provinces. Microcystin were tested for all samples by ELISA. The results show: microcystin were not detected in 12 tap water samples, were detected in 6 of 9 under water samples, 8 of 12 surface water samples; microcystin concentration of water samples from culturing pool was 207.9 pg/ml and Spires slurry was 31.9 ng/g, the former is 153 times as the latter; powder of Spires raw material was 206.4 ng/g. The average concentration of microcystin of all Spires health food samples from market was 317.2 ng/g, there into, microcystin was 142.7 ng/g in a tables and 222.6 ng/g in capsule, respectively. The results suggested: we shouldn't ignore the risk of exposuring microcystin when consuming Spires health food, it is necessary to further study and set a standard of limited microcystin in Spires health food. PMID- 14535097 TI - [Effect of apoptosis induced by different vitamin E homologous analogues in human hepatoma cells(HepG2)]. AB - To observe the effect of apoptosis induced by alpha-tocopherol(alpha-T), gamma tocopherol(gamma-T), delta-tocopherol(delta-T), and vitamin E succinate (VES) in human hepatoma cells (HepG2), apoptosis was detected by flow cytometry (FCM) and DNA ladder with different VE homologues analogues (alpha-T, gamma-T, delta-T and VES) and different concentrations (12.5 mg/L, 25 mg/L, 50 mg/L, 100 mg/L and 200 mg/L) at 48 h. The growth of HepG2 cells was inhibited by delta-T and VES at 12.5 200 mg/L in comparison with the negative control group, while (-T showed weak effect of inhibition and alpha-T did not show any inhibition effect. However, as an exception, alpha-tocopherol, delta-tocopherol and VES were effective in induction of apoptosis in HepG2 cells at concentrations of 12.5-200 mg/L. gamma tocopherol showed effect only at 200 mg/L. Conclusions delta-tocopherol and VES lowered HepG2 cells growth and viability, and increased apoptotic propensity significantly. A dose-dependent of antiproliferation and induction apoptosis was found in HepG2 cells line. The order of efficiency of four vitamin E analogues was delta-tocopherol > VES > gamma-tocopherol > alpha-tocopherol. The difference in nature and magnitude of the anticancer effects did not correlate with their reported antioxidant activity and this might be due to minor differences in their structure important to their biological activities. The results from this study suggested that delta-tocopherol and VES could be the promising anti-hepatoma agents. PMID- 14535098 TI - [Study on the safe edible of puffer fish resources]. AB - In order to utilize the puffer fish resources furthest and to develop traditional diet culture, a project on the safe eating of puffer fish resources was conducted. The results showed that the distributing of teterodotoxin(TTX) in puffer body was not in uniformity. Owing that the content of TTX changes in different seasons and bodies, different methods were selected to crank out the delicious globefish food. The number of globefish food consumer has reached the amount to 121239 person-time since 1998. The technological process on teterodotoxin was proved reliable by mice bioassay and feeding trial in human. PMID- 14535099 TI - [Study on the toxicological effect of chloropropanols on rats]. AB - Toxicological effect of 3-chloro-1,2-propanediol on rats were studied to provide scientific basis for assessing the effect of Chloropropanols on human health. 170 SD rats were divided randomly into 8 groups and the dose of 0, 0.25, 0.5, 1.0, 2.0, 4.0, 8.0, 16.0 mg/kg 3-chloro-1,2-propanediol were given to rats for 90 days by gavages per day, respectively. The weight and food efficiency, hematology and clinical chemistry, NAG, GGT and total protein in urine, sperm number, sperm survive rate and sperm aberration rate, the LDH and LDH-X activity in testis, rate of organ/weight and histopathological analysis were measured. The results showed that different dose of 3-chloro-1,2-propanediol did not has adverse effect on body weight, food efficiency, Hb, red cell, white cell, serum AST, ALT, creatine, ALP, LDH, total protein and albumin, urine GGT and total protein, LDH activity in testis. At the dose of 4.0, 8.0 and 16.0 mg/kg group, the activity of NAG in urine and the rate of kidney/weight was significantly increased compared with negative control groups; the pathological changes in kidney were observed in the same groups, and the sperm number was also significantly decreased. At the dose of 8.0 and 16.0 mg/kg group, sperm survive rate and the X-LDH activity were significantly decreased and pathological changes were also observed in testis and caudal epididymis. It was concluded that the activity of NAG in urine and sperm number is the sensitive biological effective marker. Because urine is a kind of convenient available biological material, NAG activity in urine is a good biological effective marker for assessing effect of Chloropropanols on health. If the NAG activity can be used as sensitive marker for assessment on human health need to be tested further in human study. PMID- 14535100 TI - [Effect of chronic intake of dietary fiber complex on the intestinal structure and function in hypercholesterolemic rats]. AB - To investigate the long-term effect of dietary fiber complex (DFC) on intestinal structure and function in hypercholesterolemic rats, 60 healthy SD rats were feed with food rich in lipids and hypercholesterolemic animal models were established. The animals were randomly divided into 5 groups. Rats were fed DFC at levels of 4%, 16%, or 64% for three month in the experimental groups. Wheat fiber was used in the hypercholesterolemic control (HC) group and rats feeding on normal food were used as normal control (NC). Morphology of the small intestine, reticum and caecum were observed by light and electron microscope examination. Intestinal function was measured physically. The results showed that (1) compared with NC group, fecal weight was significantly raised in DFC group of higher level (group D and E, P < 0.05); (2) the weights of small intestine wall in D and E group were significantly higher than those of NC and HC group and weights of caecum wall in E group were significantly higher than those of NC and HC group (P < 0.05); (3) widen villi and thickened muscle layer of small intestine were observed in DFC group of higher level. No demonstrable changes in reticulum morphology in any group of animals were found under the observation of light microscope (4) microvilla becoming short and/or absent, mitochondria swelling, impairment of the integrity of the cristae were commonly observed in DFC groups. Conclusions Long term intake of DFC composed mainly of Hippophae rhamnoides L, Bran, oat bran and guar gum at higher levels might induce some morphological changes of intestine and caecum. Therefore, DFC might be used at low level as an effective cholesterol lowering agent. PMID- 14535101 TI - [Effects of soybean isoflavone and calcium on proliferation, differentiation and mineralization of cultured osteoblasts in vitro]. AB - To explore the effects of soybean isoflavone and calcium on proliferation, differentiation and mineralization of cultured osteoblastic cells. Osteoblastic cells from newborn rat calvarial was cultured. Osteoblastic cells proliferation was measured by MTT, activity of APL was observed with Golden's method and ARS was used to measure the mineral nodes of osteoblasts. The results showed that the calcium group had not different from the control group on stimulating the proliferation, differentiation and mineralization of rat calvarial osteoblastic cells. The soybean isoflavone group had significant effects on stimulating the proliferation, differentiation of osteoblasts and increase the forming of the mineral nodes of osteoblasts in rat. Calcium and soybean isoflavone group had more benefit for forming mineral nodes. The soybean isoflavone stimulates the proliferation, differentiation and mineralization of rat calvarial osteoblastic cells, and increase calcium in the same time is more benefit for mineralization. PMID- 14535102 TI - [Study on the relationship between body mass index and risk factors of chronic diseases of Beijing urban residences]. AB - This study aimed to analysis the relationship among body mass index (BMI) and waist circumference (WC) with chronic diseases risk factors--serum total cholesterol (TC), triglyceridaemia(TG), high density lipoprotein cholesterol (HDL C), fasting blood glucose (FBG), systolic blood pressure (SBP), diastolic blood pressure (DBP) and the prevalence of chronic diseases. The data was collected from 1995 to 1998 in the study of community base integrated intervention of non communicable chronic diseases. RESULTS: The prevalence of overweight and obesity in male was 41.7% and 14.6%, in female was 32.0% and 20.4% in this population, respectively. BMI and WC had significant positive relationship with TC, TG, SBP and DBP but negative with HDL-C and HDL-C/TC. WC had significant positive relation with FBG. The prevalence of dyslipidaemia hypertension and diabetes were significantly increased in overweight and obese groups. For early prevention, we have to pay more attention to the population with BMI > 24.0 kg/m2, they had increased risk factors of non-communicable chronic diseases. PMID- 14535103 TI - [Prevalence survey of smoking pattern among peasants in China]. AB - To study smoking patterns among farmers so as to provide scientific basis for developing effective tobacco control measures via policy making. The Survey was conducted by randomly selecting, in multi-stage, about 50 thousand farmers aged over 15 years old as the participants of the study according to economic status and geographical distribution in Guangdong, Henan, Jiangsu, Qinghai and Heilongjiang provinces. Results The overall prevalence of smoking was 33.45% (prevalence in men was 56.94% and in women was 6.40%) showing a decrease of 5%, 10% and -2.5% as compared with those of in 1996 survey. The average number of cigarettes consumed per day by men and women was 16 and 13 respectively as compared with 15 and 11 in 1996. CONCLUSION: Smoking prevalence among farmers in China was still remarkably high. The measures for tobacco control in rural areas should be emphasized. For reducing this prevalence, health promotion strategy might be prioritized. PMID- 14535104 TI - [Study on efficacy of adult obesity after intervention in community, Anyang]. AB - To evaluate the efficacy of a community-based adult obesity intervention in Angang. The intervention included health education activities among residents in a community in Anyang. 2400 and 1200 individual were randomly selected as intervention group and control group respectively from a community in Anyang. The questionnairing survey and body weight measuring were conducted and repeated at the end of the study for one year. The results showed that the awareness on the prevention of obesity was enhanced(P < 0.05) and mean weight of target population decreased. The community-based intervention for obesity was effective as well as feasible. PMID- 14535105 TI - [Toxicity of tetrodotoxin towards mice and rabbits]. AB - The toxicity of tetrodotoxin(TTX) in Kunming mice and rabbits was investigated in this study. The median lethal doses (LD50) were 10.7, 12.5, 532 micrograms/kg for intraperitoneal (i.p.), subcutaneous (s.c.) and intragastric (ig) administration of toxin respectively in mice and, it was found that the male mice was more sensitive to TTX than the female was. TTX toxic doses for both intramuscular (i.m.) and intravenous (i.v.) administration via injection in rabbits were reported first time and the minimal lethal doses(MLD) were 5.3, 3.1 micrograms/kg and wholly lethal doses(LD) were 5.8, 3.8 micrograms/kg for i.m. and i.v. injection respectively. TTX-intoxication symptoms and characteristics were described in both animal species and in different administrations of toxin. TTX was found to be about fifty times less toxic and to have more delayed death occurrence to mice via oral route than that via i.p. injection, providing, more likely, a favorable potential for the cure of actual TTX-poisoning victims. PMID- 14535106 TI - [Determination of dissociate formaldehyde in cosmetics by derivative gas chromatography analysis]. AB - A method for determination of dissociate formaldehyde in cosmetics by derivative gas chromatography analysis was presented in the paper. The lowest detection limit of analysis was 5.75 pg; the linear calibration curve for formaldehyde were measured within the range of 8.3-166 micrograms/L, and the correlative coefficient was 0.9996. It was showed that this method was simple and easy to handle, and interferential factors were less, and accuracy was high. PMID- 14535107 TI - [Study on the effect of NaFeEDTA on preventing lead poisoning in rats and its impact on zinc, copper]. AB - In order to explore the effect of sodium iron ethylenediaminetetraacetate (NaFeEDTA) on preventing rats from lead poisoning and reducing the lead toxicity, and at the same time to explore its impact on zinc and copper, sixty Wistar rats were randomly divided into six groups including blank control group, model control group, positive control group and three different level NaFeEDTA (high, middle and low) groups. With the exception of the blank control group, which only received double distilled water treatment throughout the experiment, all the other rats were continuously treated with lead solution. At the same time, with the exception of the blank and model control groups, the other four groups were treated with acidi dimercaptosuccinici (DMSA), high, middle and low level NaFeEDTA solutions respectively. At the end of the study, several indicators were determined. The results showed that the hemoglobin levels in the three different level NaFeEDTA groups were significantly higher than those of the other four groups and at the same time the lead levels in blood and organs were lower than those of model group significantly. There were significant reverse correlations between iron supplement and the lead levels in liver, kidney and tibia among the three different level NaFeEDTA groups, and the correlation coefficients were 0.4432, -0.6134 and -0.3878 respectively. There were no significant differences in zinc levels and copper levels between the three level NaFeEDTA groups and the negative control group except the brain zinc levels in the middle and high level NaFeEDTA groups were lower and the kidney copper level in the low level NaFeEDTA group was higher than those of the model control group. It is concluded that the NaFeEDTA can effectively improve hemoglobin level, prevent rats from lead poisoning and reduce the lead toxicity. At the same time, NaFeEDTA does not have significant influence on zinc and copper levels in rats. PMID- 14535108 TI - [Study of ceratocystis fimbriata toxins determination from sweet potato by high performance thin-layer chromatography]. AB - The extract solution from Sweet Potato was washed with 5% Na2CO3 and followed with distilled water and dried with dehydrated sodium sulfate (Na2SO4). The ether extract was evaporated and the residue (crude sample) was dissolved in a small volume of chloroform-soluble solution and was spotted on a silica gel G thin layer plate developed with different solvent systems. Pure ipomeamarone and ipomeamaronol were used as standards. The resolution was the best in the solvent system of petroleum ether-ethylacetate at 20 degrees C. The separated spots were scanned by CS-930 TLC scanner at 520 nm and quantitative determination by external standard calibration. Ipomeamarone was detected by high performance thin layer chromatography and the limits of detection were 0.002 microgram for pure and 0.01 microgram for crude sample. PMID- 14535109 TI - [Analysis of kojic acid in Aspergillus oryzae ferment by ion-pair reversed-phase high performance liquid chromatography]. AB - A new and an efficient method for analyzing kojic acid in aspergillus oryzae ferment by ion-pair reversed-phase high performance liquid chromatography was developed. The kojic acid was well separated on a C18 column using methanol-0.01 mol/L disodium hydrogen phosphate and 2 mmol tetrabutylammonium bromide (4:96) as the mobile phase, followed by an UV detection at 226 nm. The detection limit was 0.012 microgram/ml. The spiked recoveries in wort-yeast culture, wort-peptone culture and potato-yeast-sugar culture were 98.2%, 103.4% and 97.2%, respectively. The RSD values were 0.51%, 0.45% and 0.43% respectively. PMID- 14535110 TI - [Prediction formulas for estimating body fat percent of obesity from bioelectrical impedance]. AB - The bioelectrical impedance method has become a popular and widely used method to determine body composition. In this study, body fat was measured by densitometry in a group of 66 males and 130 females obesity Chinese, aged 18 to 67 years. Body weight, body height were measured as well as total body bioelectrical impedance at 50 kHz. BMI and BF% was (27.7 +/- 3.1) kg/m2 and (38.81 +/- 6.27)% respectively (mean +/- SD). The prediction formulas for body fat from bioelectrical impedance developed in western societies, are significant underestimate body fat of Chinese obesity from body density. The prediction formulas for body fat with our subjects is; BF = 0.846 * Wt - 0.185 * Ht2/Z - 2.361 * Sex - 24.977. (r = 0.923, SEE = 3.43, F = 365.7, P < 0.001). Wt: body weight; Ht: body height; Sex; male = 1, female = 0; Z: bioelectrical impedance. PMID- 14535111 TI - [Simultaneous spectrophotometric determination of mixed colorants by artificial neural network]. AB - The back propagation artificial neural network was applied to process spectrophotometric data of mixed colorants. By optimizing the network structure and parameters, the predication accuracy was enhanced. The method was used for simultaneous determination of mixed colorants in synthetic samples and drinks on sale with satisfactory results. Therefore, the artificial neural network may provide a new approach to determine the mixed colorants is drinks by spectrophotometry without any preliminary chemical separation. PMID- 14535112 TI - [Rapid determination of formaldehyde in food packaging material by oscilloscopic polarography]. AB - The polarographic behavior of the reaction product of formaldehyde and phenyldrazine hydrochloride on the drop mercury electrode was investigated in a medium of 0.01 mol/L H3PO4 and sensitive second order polarographic reduction wave was obtained at -0.76 V (vs. SCE). The optimum conditions and disturbance of other co-existing substances were also studied, and were compared with those of national standard method. In the study, the detectable range, detection limit, recovery rate, relative standard deviation(RSD) of the improved method were 0.005 0.25 mg/L, 0.005 mg/L, 92.3%-104.6%, 1.1%-3.0% respectively, revealing the accordance with the national standard method. PMID- 14535114 TI - [Advance in di-n-butyl phthalate]. AB - Di-n-butyl phthalate is ubiquitous because it is widely used in plastic and cosmetic production. In this paper, the research progress on the exposure, metabolism, toxicity and endocrine disrupting effects of di-n-butyl phthalate were reviewed. PMID- 14535113 TI - [Genetic polymorphism and risk of diseases induced by chemicals]. AB - The response of the individuals to xenobiotics is various due to the genetic polymorphism, that is, most likely, the determinant factor of the susceptibility of the individuals to xenobiotics. Therefore, the risk of diseases taking cancers as an example is different in population. In this paper, progresses of genetic polymorphism of toxicant-metabolizing enzyme genes and DNA repair genes and, their effects on individuals' risk to diseases were briefly reviewed. PMID- 14535115 TI - [Studies on the effects of chromium complexes]. AB - The existence of chromium complexes in nature, their industrial uses, and their health impacts were reviewed in this paper. The toxic and the formation of Cr(V/IV) complexes and the mechanisms of DNA damage were discussed in particular. The major damaging species responsible for the genotoxicity were Cr(V) or Cr(IV) complexes. However, the function of reactive organic intermediates could not be excluded. PMID- 14535116 TI - [Toxicogenomics--the research of environment and health at gene expression level]. AB - Toxicogenomics is a new scientific field that elucidates how the entire genome is involved in biological responses of organisms exposed to environmental stressors and provides profound understanding of the roles of gene-environment interactions in pathogenesis of disease using high throughput technologies. In this paper, the research progress on the toxicogenomics and its application in both improving environmental health and preventing environmentally related disease were reviewed. PMID- 14535117 TI - [Factors of antioxidant and prooxidant activities of carotenoids]. AB - The effectiveness of carotenoids as antioxidants is dependent on a number of influencing factors. It is likely that carotenoids exhibit a tendency to lose their effectiveness as antioxidants or act as prooxidants. In this paper, factors influencing the antioxidant or prooxidant activities of carotenoids such as the molecule structure of carotenoids, the location or site of action of the carotenoid molecule within the cell, the concentration of carotenoids, the properties of reactants, the partial pressure of oxygen and the interaction with other antioxidants were reviewed. PMID- 14535118 TI - Serious business. While physicians are busy seeing patients, practice managers drive the financial success of the operation with information technology. PMID- 14535119 TI - Scrip for success. Kentucky family practice uses electronic prescriptions to improve efficiency, revenue and customer service. PMID- 14535120 TI - Calling for automation. Pediatric call center at a Missouri children's hospital sees lower call times and higher productivity from a fully automated system. PMID- 14535121 TI - Converting interim HIPAA fixes into long-term strategies. HIPAA still represents an opportunity for MCOs to strengthen their financial efficiency, but only if they employ a strategic approach. PMID- 14535122 TI - New promise for interorganizational collaboration. As a network platform, VPNs can help beat the top five obstacles healthcare organizations face in trying to initiate and sustain information-sharing networks. PMID- 14535123 TI - What works. Taking it to the next level. Georgia specialty practice provides greater care by improving efficiency with EMR system. PMID- 14535124 TI - Security considerations when choosing an EMR system. PMID- 14535125 TI - What works. Paper-plagued to paperless. Florida healthcare system now has instant access to patient charts anytime and anywhere it has Internet access. PMID- 14535126 TI - Practice management systems hotlist. PMID- 14535127 TI - Patient participation in EHR benefits. AB - Patient participation in EHR creation can significantly broaden the range of healthcare information in the record and, ultimately, improve clinical decision support. The care with which patient-authored information is handled, however, must equal the care devoted to clinician data entry and review. Web-based access, self-service management, consent delegation and agent permissions management are just a few of the features that will encourage active and positive patient involvement with their healthcare records and with their own health management. PMID- 14535128 TI - Consumer-driven health care: transforming medical management. PMID- 14535129 TI - Staging: a revolution in helping people change. PMID- 14535130 TI - Managing dyslipidemia in adults with diabetes. PMID- 14535131 TI - Prevention: providing lipid management in a large clinical practice. PMID- 14535132 TI - Patient-focused care boosts patient satisfaction while enhancing safety. AB - Bringing treatment to patients lessens their pain and suffering. Services delivered at point of care include monitoring and testing. New technology helps avoid handoffs of patients and information. PMID- 14535133 TI - Checkoffs play key role in SICU improvement. AB - Unit sees 25% drop in mortality following implementation of new form. Checklist not a to-do list, but enumeration of clinical, safety goals. All members of the multidisciplinary team are involved in process. PMID- 14535134 TI - Program distinguishes illness from disease. AB - Student teams will pay eight home visits in two-year program. Topics will examine the impact of disease on patients and their family members. First-year med students form beliefs that will last their whole career. PMID- 14535135 TI - HFAP survey earns facility referral status. AB - Accreditation alternatives to the Joint Commission on Accreditation of Health Care Organizations (JCAHO) are not necessarily an either- or proposition. The American Osteopathic Association is seen as more document-driven, while JCAHO focuses on outcomes. Consultants prove invaluable in helping center prepare for Healthcare Facilities Accreditation Program survey. PMID- 14535136 TI - IOM cites core functions to improve EHR systems. AB - Key elements must address quality, efficiency, and feasibility. Committee piggybacks on ongoing study for faster results. Common industry standard for software developers being created. PMID- 14535138 TI - Patient safety alert. Study shows safety risks for children, value of tool. PMID- 14535137 TI - Patient safety alert. Hospital safety initiative targets behavioral health unit. PMID- 14535139 TI - Professional development: getting the most from the summit. PMID- 14535141 TI - Faculty matters. Jennifer Couvillon. PMID- 14535142 TI - A snapshot of nursing in Qatar. AB - Between September 2002 and February 2003, shortly before the military action in Iraq. Virginia Nehring served on the faculty of the College of Science in Doha, Qatar. For her second experience as a Fullbright scholar--she taught nursing in Africa in 1994--Dr. Nehring sought to experience an entirely different region of the world, one where English would be spoken and understood. When she made her decision to work in Qatar, a small peninsula nation off the coast of Saudi Arabia, the country was not well known to Americans. Now it is known as Central Command headquarters for the American military during the Iraq military action and as the home of Al Jazeera, the influential television news and opinion station for the Middle East. During her stay in Qatar, Dr. Nehring became the fourth, and only non-Egyptian, doctorally prepared faculty person in the nursing unit of the College of Science. Besides teaching courses such as ethics and issues and trends, she carefully assessed the nursing program, much as an accreditation site visitor might do, with feedback to university administration as well as the nursing unit. She left the region shortly before the military action began. PMID- 14535143 TI - "That spirit, that thing inside". Using qualitative research techniques to produce a recruitment film for Hispanic/Latino and American Indian students. AB - This article reports on the use of qualitative research methodology in producing a 23-minute recruitment film to attract Hispanic/Latino and American Indian students into a baccalaureate nursing program. The research question addressed in this pilot project was, "What is the meaning of the educational experience to Hispanic/Latino and American Indian nurses in the Yakima Valley of south central Washington State?" The conceptual development of the project, recruitment of interview participants, generation of interview protocol, data collection, and analysis are described and correlated to accepted qualitative research elements. Themes and patterns identified in participant interviews, the "findings" of the project, were used during the postproduction phase as guides for the story that unfolds in the film, which addresses the experience of being a nurse of color. PMID- 14535144 TI - Analyzing the teaching style of nursing faculty. Does it promote a student centered or teacher-centered learning environment? AB - The purposes of this study were to a) describe the predominant teaching style of a group of nursing faculty members, either as teacher centered or student centered, and b) to compare teaching style to the instructional methods the faculty members used in the courses they taught and to their stated philosophies of teaching/learning. Findings indicate that the participants were more teacher centered than student centered; their written philosophies supported the teacher centered approach. However, evidence that faculty used student-centered language, often in a teacher-centered context, indicates that participants in the study may recognize the need for a student-centered environment but may have difficulty with implementation. Recommendations for faculty members and administrators are offered. PMID- 14535145 TI - Academic performance of nursing students: do prerequisite grades and tutoring make a difference? AB - The purpose of this study was to evaluate the relationship of prerequisite science course performance and tutoring services with academic performance of first semester nursing students. Quantitative data were gathered from student records after a semester in which tutoring services were offered. The convenience sample consisted of 37 students enrolled in first semester nursing courses. Influence on academic performance was determined by comparing the academic performance of two groups, students who attended four or fewer tutorial sessions and students who attended five or more tutorial sessions. Previous academic performance was shown to have a statistically significant relationship with first semester nursing school performance. Recommendations for nursing faculty are offered. PMID- 14535146 TI - A collective case study of nursing students with learning disabilities. AB - This collective case study described the meaning of being a nursing student with a learning disability and examined how baccalaureate nursing students with learning disabilities experienced various aspects of the nursing program. It also examined how their disabilities and previous educational and personal experiences influenced the meaning that they gave to their educational experiences. Seven nursing students were interviewed, completed a demographic data form, and submitted various artifacts (test scores, evaluation reports, and curriculum based material) for document analysis. The researcher used Stake's model for collective case study research and analysis (1). Data analysis revealed five themes: 1) struggle, 2) learning how to learn with LD, 3) issues concerning time, 4) social support, and 5) personal stories. Theme clusters and individual variations were identified for each theme. Document analysis revealed that participants had average to above average intellectual functioning with an ability-achievement discrepancy among standardized test scores. Participants noted that direct instruction, structure, consistency, clear directions, organization, and a positive instructor attitude assisted learning. Anxiety, social isolation from peers, and limited time to process and complete work were problems faced by the participants. PMID- 14535148 TI - The pursuit of excellence in nursing education. AB - The National League for Nursing is clear in its mission and its commitment to providing leadership in re-creating the landscape of nursing education. This commitment is grounded in the certainty that excellence in nursing education can be achieved. The initiatives described here are the newest and most exciting programs advanced by the NLN. We look forward to your joining us in this multifaceted effort to advance excellence in all types of nursing programs, and we ask you to remember the words contained in a contemporary piece of advice as you face the challenges inherent in this work: Excellence can be attained if you care more than others think is wise, risk more than others think is safe, dream more than others think is practical, and expect more than others think is possible. PMID- 14535147 TI - An innovative strategy to enhance new faculty success. AB - Faculty stress is a factor in the current shortage of nursing faculty. New faculty members in schools of nursing are subject to stress from many sources. This article reports on an innovative strategy that decreases stress for new faculty while facilitating faculty tenure success. PMID- 14535149 TI - Implant surface design: using biotechnology to enhance osseointegration. Interview. PMID- 14535150 TI - A concept analysis of the role of fatherhood: a Japanese perspective. AB - A concept analysis of the father's role is presented from a Japanese perspective. To conceptualize the term father's role, the process for concept analysis was employed. The analysis resulted in a definition of the term and four defining attributes. Two dimensions of the father's role emerged as being most commonly used or implied: a behavioral dimension and a consequential dimension. Each of these dimensions had another four subcategories in terms of the direction of the consequences: those were to the child, the mother, the family, and the father himself. The findings will assist in instrument development and to a further understanding of the concept that will stimulate nursing theory development and research. PMID- 14535152 TI - A study of resilience in young Ethiopian famine survivors. AB - We studied resilience related to childhood experiences of famine in Ethiopia. We sought an understanding of how young Ethiopians survived and coped with the devastating effects of famine. Participant observations and in-depth interviews at an orphanage in Addis Ababa with eight boys and girls (ages 18 to 23), survivors from famine catastrophe in Ethiopia 1984-1985, were conducted. Significant resilience factors identified were faith and hope, having a living relative, and having memories of one's past roots. Exposure to famine and multiple early losses may have long-term effects on an individual's capacity to maintain resilience. We discuss how an understanding of resilience can be applied in different cultural settings. Future research on resilience in Africa is suggested to increase our knowledge base of this concept. PMID- 14535151 TI - Culturally interpreting environment as determinant and experience of health. AB - In minority communities, experiences of the environment and health disparities are frequently related. An important component of nursing practice in communities is to address these issues in cultural context to improve overall community health experiences. This present ethnographic study was conducted in an urban barrio community in which residents have responded to health threats posed by local environmental hazards with sustained community-focused health and development strategies. Data generation occurred during field visits in the community and included interviews, participant observation, field notes, participant-drawn maps, examination of artifacts and existing documents, and photography. Two dimensions define the community experience of the environment in this community: contamination and unhealthy community. The environment is a complex and multidimensional concept of central importance to community experiences. Environmental health cannot be addressed without first understanding that experience in cultural context. PMID- 14535153 TI - Symptoms experienced during menopausal transition: Korean women in South Korea and the United States. AB - This article reports on cultural influences on symptoms experienced during menopausal transition of Korean women in South Korea and Korean immigrant women in the United States. Data from independent studies of two groups of Korean women were triangulated and analyzed using descriptive and inferential statistics. The analysis indicated that Korean women in South Korea tended to report more symptoms than Korean immigrant women in the United States. Types and severity of prevalent symptoms were also found to be different between the two groups. The findings suggest that recent introduction of menopausal industries in South Korea and contextual influences on Korean women's work and immigration in the United States would be the reason for differences. Based on the findings, implications for future research are proposed. PMID- 14535154 TI - The effect of social support and acculturation on postpartum depression in Mexican American women. AB - Postpartum depression was studied in 66 multiparous Mexican American women using a prospective longitudinal design. Interviews were conducted at 34 to 36 weeks prenatally and 4 to 6 weeks postpartum. Women expressing prenatal depression were more likely to continue to experience depression postpartum. Other predictors included a high need for postpartum support, specific support network characteristics, acculturation, and poor quality relationships within the family. It is important for the health care provider to assess the presence of depressive symptoms and available social support during the prenatal period and work with Mexican American mothers to increase support and secure help in dealing with daily life stressors. PMID- 14535155 TI - An exploration of the experiences of cultural safety educators in New Zealand: an action research approach. AB - This research is a study of the experiences of four cultural safety educators in nursing education in Aotearoa, New Zealand. Action research methods assisted the participants to implement change in their practice and gain positive personal involvement in the study. Reflective diaries provided the major tool in this process as participants were able to achieve at least one action research cycle by identifying issues, planning action, observing the action, and reflecting. The findings of the research revealed that the participants not only coped with everyday stressors of teaching but were also required to formulate knowledge for cultural safety. For the Maori participants, their stress was confounded with recruiting and retaining Maori students and macro issues such as commitment to their iwi (tribe). Lack of support to teach cultural safety was identified to be a key theme for all participants. An analysis of this theme revealed that many issues were organizational in nature and out of their control. Action research provided a change strategy for participants to have a sense of control of issues within their practice. Recommendations include the following: support for cultural safety educators to have a dialogue on a regular basis, the introduction of nurse educator programmes, paid leave provisions for educators to conduct and publish research so that a body of knowledge can be developed, and that Maori cultural safety educators be recognised for their professional and cultural strengths so that they do not fall victim to burnout. PMID- 14535156 TI - A transcultural ethos underpinning curriculum development: a master's programme in disaster relief nursing. AB - This article discusses the development and implementation of a masters' degree programme in disaster relief nursing by cooperating partners in European countries. It aims to enhance the provision of aid by endorsing the role of nurses at the forefront of health care in disaster-stricken areas around the world. As an attempt to address the implications of globalization in health care, the programme was established as a recognition of the rapidly developing specialty of transcultural/transnational nursing. Innovative teaching and learning strategies used include e-learning, simulation exercises, and competency portfolio development. A rigorous evaluation of programme outcomes is ongoing. PMID- 14535157 TI - Linguistic services in ambulatory clinics. AB - A review of the literature reveals few studies that focus on the challenge of language barriers in primary care settings. Recognizing the need for a national consensus on cultural and linguistic standards for health care in the United States, the Office of Minority Health recently released a set of standards for culturally and linguistically appropriate services (CLAS). These standards were utilized to examine the linguistic services available at eight ambulatory care centers in a small New England state in an effort to determine compliance with recommended national standards. Although myriad studies have focused on provision of linguistically appropriate care in emergency rooms (ERs), few studies have specifically examined ambulatory care settings. Numerous strategies have been adopted by individual clinics in an attempt to deal effectively with linguistic barriers. Yet without clear national regulations and dedicated funding for interpreter services, a large spectrum of services exists. Survey data were obtained from on-site visits at select community health clinics to ascertain availability, need, and utilization of linguistic services for patients with limited English proficiency. The majority of patients served by the clinics surveyed were predominantly Spanish-speaking. Results reveal that although most of the clinics provided informal mechanisms of interpreter services, few directly addressed linguistic services as a component of culturally competent care. PMID- 14535158 TI - [Human activities and American trypanosomiasis. Review of the literature]. AB - Trypanosoma cruzi, the causative agent of American trypanosomiasis (Chagas disease) is a parasite of wild mammals of Americas. It has been further transmitted to man through an insect vector belonging to the Reduviidae family. In the context of an ecoepidemiological study of this vector borne anthropozoonosis, a medical anthropological overview tries, by using data from literature, to clarify the different ways human activities play on the disease epidemiological cycle. The everyday human activities support domiciliation of triatomine bugs. But, the human intervention in the natural environment disturbs strongly the ecology of the vectors or hosts and leads to a new structuring of the natural foci of the disease with its displacement toward the human environment. The way society or individuals manage the disease plays also an important role. Some human activities, as building dwelling structures, have been well studied, but some others, such as the real impact of the human modes of production, are less known. A medical anthropology study of the role of human activities on the foci of Chagas disease is still needed. PMID- 14535159 TI - Pseudophysaloptera enigmatica sp. nov. (Nematoda: Physalopteridae) from Dactylopsila trivirgata Gray, 1858 (Marsupialia: Petauridae) in Australia, with a review of the genus. AB - The nematode Pseudophysaloptera enigmatica sp. nov. is described from the stomach of Dactylopsila trivirgata Gray, 1858, in Queensland, Australia. The new species is distinguished from congeners by its size and features present on the tail of the male. P. enigmatica sp. nov. is considerably larger than virtually all congeners, and can be differentiated by the presence of spicules and the shape and distribution of caudal papillae in the male tail. This is the first record of the genus in Australia and in marsupials. A review of the genus is provided. PMID- 14535161 TI - Litomosoides yutajensis n. sp., first record of this filarial genus in a mormoopid bat. AB - Twenty-five bats were trapped in Yutaje (Amazonas, Venezuela) and examined for Litomosoides (Filarioidea: Onchocercidae). Of the nine recovered bat species, only Pteronotus parnelli was infected; it is a cave-dwelling species belonging to a family, Mormoopidae, which has not previously been included in the host range of the genus. The new species, L. yutajensis n. sp., has two median cephalic bosses covered with rugosities and differs from the 15 recognized species and subspecies from bats in several characters. Alike L. molossi Esslinger, 1973, L. chandleri Esslinger, 1973 and L. chitwoodi Bain, Guerrero, Rodriguez 2003, the new species has cuticular lateral bosses on the body. Eight of 10 P. parnelli were microfilaraemic, but only three had adult worms, showing that microfilariae survive longer than adults, which could lengthen the period of transmission. No infective larvae were detected in the following macronyssid mites: 58 Ornithonyssus bacoti, Ornithonyssinae, experimentally fed on microfilaraemic bats and dissected 15 days later, and a few Radfordiella sp., Macronyssinae, recovered from P. parnelli. PMID- 14535160 TI - Examination of type material of two species of Litomosoides (Filarioidea: Onchocercidae), parasites from bats; taxonomic consequences. AB - Type material of Litomosoides hamletti Sandground, 1934 from Glossophaga soricina soricina in Brazil and L. penai Jimenez-Quiros & Arroyo, 1960 from Carollia perspicillata azteca in Costa Rica, was examined. The morphology of the spicules shows that these species belong to the carinii group. Their synonymy with L. guiterasi Perez Vigueras, 1934, from Artibeus jamaicensis yucatanicus in Cuba, does not appear justified because they are distinct in several characters (body length, width of female, size and shape of buccal cavity and capsule, shape of right spicule). L. hamletti is a valid species; L. penai is closely related to it and is considered to be a sub-species, L. hamletti penai Jimenez-Quiros & Arroyo, 1960. The material recovered from Glossophaga spp., previously assigned to L. guiterasi by several authors, is identified as L. h. hamletti. L. guiterasi appears to be closely related to L. chandleri Esslinger, 1973; L. chitwoodi n. sp. (= Litomosoides sp. Chitwood, 1938) seems close to these species; all three are parasites of Artibeus spp. PMID- 14535162 TI - Corynosoma acanthocephalans in their paratenic fish hosts in the northern Baltic Sea. AB - Cystacanth stages of three Corynosoma (Acanthocephala) species, C. strumosum and C. semerme, and a new species in the Bothnian Bay, C. magdaleni, were studied in the fourhorn sculpin (Myoxocephalus quadricornis) in 1996-1997. The length of the trunk and proboscis differentiated the three species in the fish. The stability of the Corynosoma infection was studied by comparing the present results with those collected from the same areas (central and coastal) in 1977-1982 (Valtonen, 1983a). As C. magdaleni and C. strumosum were not separated at that time, the joint infections of these two species (called "C. strumosum") were compared. The stability of infection with C. semerme in the central Bothnian Bay was noticeable (prevalences were 82.9 in earlier period and 81.9% in later period), while the prevalences of "C. strumosum" in the same areas had decreased (21 and 13.5%, respectively). This is suggested to be due to the disappearance of the marine bull-rout, Myoxocephalus scorpius, from the Bothnian Bay during the 1990s due to an overall decline in salinity throughout the Baltic Sea. In the coastal area Corynosoma infection was clearly lower than in central area in both periods. PMID- 14535163 TI - Rileyella petauri gen. nov., sp. nov. (Pentastomida: Cephalobaenida) from the lungs and nasal sinus of Petaurus breviceps (Marsupialia: Petauridae) in Australia. AB - A new cephalobaenid pentastome, Rileyella petauri gen: nov., sp. nov. from the lungs and nasal sinus of the petaurid marsupial, Petaurus breviceps, is described. It is the smallest adult pentastome known to date, represents the first record of a mammal as the definitive host of a cephalobaenid and may represent the only pentastome known to inhabit the lungs of a mammal through all its instars, with the exception of patent females. Adult males, non-gravid females and nymphs moulting to adults occur in the lungs; gravid females occur in the nasal sinus. R. petauri is minute and possesses morphological features primarily of the Cephalobaenida but the glands in the cephalothorax and the morphology of the copulatory spicules are similar to some members of the remaining pentastomid order, the Porocephalida. This unusual combination of features distinguish the new genus from other genera in the Cephalobaenida. The occurrence of only seven fully-formed larvae in eggs in the uterus, each representing about 10% of the length of the patent female, and her presence in the nasal sinus of a dependent juvenile P. breviceps (36 gm) implies a direct life cycle. PMID- 14535164 TI - Insights into the relationships of Palearctic and Nearctic lymnaeids (Mollusca: Gastropoda) by rDNA ITS-2 sequencing and phylogeny of stagnicoline intermediate host species of Fasciola hepatica. AB - Fascioliasis by Fasciola hepatica is the vector-borne disease presenting the widest latitudinal, longitudinal and altitudinal distribution known. F. hepatica shows a great adaptation power to new environmental conditions which is the consequence of its own capacities together with the adaptation and colonization abilities of its specific vector hosts, freshwater snails of the family Lymnaeidae. Several lymnaeid species only considered as secondary contributors to the liver fluke transmission have, however, played a very important role in the geographic expansion of this disease. Many of them belong to the so-called "stagnicoline" type group. Stagnicolines have, therefore, a very important applied interest in the Holarctic region, to which they are geographically restricted. The present knowledge on the genetics of stagnicolines and on their parasite-host interrelationships is, however, far from being sufficient. The present paper analyses the relationships between Palaearctic and Nearctic stagnicoline species on the base of the new light furnished by the results obtained in nuclear rDNA ITS-2 sequencing and corresponding phylogenetic studies of the lymnaeid taxa Lymnaea (Stagnicola) occulta, L. (S.) palustris palustris (topotype specimens) and L. (S.) p. turricula from Europe. Natural infections with F. hepatica have been reported in all of them. Surprisingly, ITS-2 length and GC content of L. occulta were similar and perfectly fitted within the respective ranges known in North American stagnicolines. Nucleotide differences and genetic distances were higher between L. occulta and the other European stagnicolines than between L. occulta and the North American ones. The ITS-2 sequence of L. p. turricula from Poland differed from the other genotypes known from turricula in Europe. The phylogenetic trees using the maximum-parsimony, distance and maximum-likelihood methods confirmed (i) the inclusion of L. occulta in the branch of North American stagnicolines, (ii) the link between the North American stagnicolines-L. occulta group with Galba truncatula, and (iii) the location of the L. p. turricula genotype from Poland closer to L. p. palustris than to other European L. p. turricula genotypes. The Palaearctic species occulta is included in the genus Catascopia, together with the Nearctic species catascopium, emarginata and elodes. The results suggest a potential of transmission capacity for C. occulta higher than that of other European stagnicolines or Omphiscola glabra. The relatively low genetic distances between C. occulta and G. truncatula and the clustering of both species in the same clade suggest that C. occulta may be potentially considered as the second lymnaeid intermediate host species of F. hepatica in importance in eastern and northern Europe, and probably also western and central Asia, after G. truncatula. L. p. turricula may be considered as a potential secondary vector of F. hepatica, at a level similar to that of L. p. palustris. PMID- 14535165 TI - The life cycle and seasonal changes in the occurrence of Pomphorhynchus laevis (Palaeacanthocephala, Pomphorhynchidae) in a small isolated lake. AB - In a small isolated lake in Slovakia, the fish acanthocephalan Pomphorhynchus laevis using Gammarus balcanicus and the minnow Phoxinus phoxinus, respectively, as its intermediate and final hosts, represented a dominant helminth species. Its prevalence and intensity of infection in fish showed no significant variation during a year fluctuating above the mean values of 89% and 6.6 worms per fish. The mean prevalence of P. laevis larvae in Gammarus was 41.4% with a maximum in the late summer and autumn; individual crustaceans were infected by 1-9 larvae. There was one generation of P. laevis per year. Following up an annual cycle, an occurrence of new infections of Gammarus culminated in October and in the next May for Phoxinus. The sex ratios of both the adults and larvae of acanthocephalans were near unity but favoured slightly males in spring and autumn. The distribution of P. laevis in minnows and crustaceans was highly aggregated and fitted with the negative binomial model. The spatial distribution analysis of parasites along the fish alimentary tract showed a clear preference of P. laevis for its proximal half, with the maximum numbers in the site of the first intestinal loop. Immature worms of both sexes predominated in the proximal region and moved slightly down the alimentary tract during their growth and maturation. PMID- 14535166 TI - Parasitological and immunological effects induced by immunization of Mandrillus sphinx against the human filarial Loa loa using infective stage larvae irradiated at 40 Krad. AB - Six mandrills were immunized with 150 Loa loa infective stage larvae (L3) irradiated with 40 Krad, and challenged with 100 L3, 60 days after initial vaccination. The parasitological outcome of this immunization was compared to results from six mandrills infected with normal L3. No clear association was seen between vaccination and microfilaremia until day 245 when a significant drop in the level of microfilaria occurred in vaccinated compared to infected animals (5 vs 10 mf/ml; p = 0.012). A one-year follow-up of the humoral immune response showed a strong adult, microfilariae (Mf) and L3 specific IgG response, with distinct profiles for each extract. In immunized animal a significant decrease in antibody level was systematically observed between days 90-145 for the anti-L3 and anti-adult IgG. However, in the same group anti-Mf antibody levels that peaked around 160-175 days post-challenge, were inversely correlated with the decrease in Mf density between day 200 and day 386. These results suggest that immunization with irradiated L3 using these specific conditions may affect the appearance of Mf. PMID- 14535167 TI - [New data on parasitization of Glossina palpalis palpalis (Diptera: Glossinidae) by Hexamermis glossinae (Nematoda: Mermithidae) in a forested area of Ivory Coast]. AB - Observations on the parasitism of Glossina palpalis palpalis by Hexamermis glossinae were carried out over a period of one year by catching flies at Abengourou, Aboisso and Daloa (forested area of Ivory Coast). No parasite is observed out of 2,168 Glossina palpalis palpalis caught in Abengourou and 9,732 in Aboisso. At Daloa, dissections of 7,341 Glossina reveal 1.75% parasited flies. All the worms were located in the abdominal cavity, loosely intertwined with the internal organ. Males were more infested than females (2.68% versus 1.26%). The parasites were more abundant among the nulliparous (2.30%) than the young parous (1.19%) and than the old parous (0.52%). The majority of infected flies were caught at the beginning of the rainy season (5.17%) and few in the dry season (0.23%). The low parasitic infection rate observed here indicates a minimal effect on the population dynamics of the vector of sleeping sickness in Ivory Coast. PMID- 14535168 TI - [Molecular characterization of mosquitoes of the Anopheles gambiae complex from Mayotte and Great Comoro]. AB - The mosquitoes of the Anopheles gambiae complex have been characterised at specific and sub-specific levels in two islands of the Comoros archipelago: the island of Mayotte (French departmental collectivity) and the island of Grande Comore (Comoros Union). Results are similar in the two islands and are presented together. The species An. gambiae s.s. was observed alone (determination performed on 149 specimens by PCR product of IGS of rDNA). The molecular form observed alone was S, and corresponds in this geographic area to the chromosomal form Savanna (determination performed on 123 specimens by another PCR product of IGS of rDNA). The haplotype IB was observed alone (determination performed on ten specimens, by sequencing the ITS of rDNA, with special attention at the position 871 of ITS), as previously observed by other authors in East Africa. Finally, in Mayotte and Grande Comore the An. gambiae complex is only composed by An. gambiae s.s. from the molecular form S/type IB. PMID- 14535169 TI - Erythrocyte antioxidative enzyme activities and lipid peroxidation levels in patients with cutaneous leishmaniasis. AB - Although cutaneous leishmaniasis (CL) is a local infection, the cellular immune response against the disease is systemic, and reactive oxygen intermediates (ROIs) are an important part of cellular immune response involved in killing the parasite. However, whether these intermediates cause oxidative damage in host is unknown. In this study, the metabolism of ROIs were investigated in patients with CL, and compared with healthy subjects. Erythrocyte lipid peroxidation was determined, as an index of oxidative damage, by measurement of malondialdehyde (MDA) levels. Erythrocyte MDA levels and SOD activities were found to be significantly higher in CL patients than in control subjects (p < 0.01 and p < 0.01, respectively). However, CAT and GSH-Px activities were significantly lower in the CL group (p < 0.05 and p < 0.01, respectively). There was a tendency to decreased hemoglobin (Hb) levels, but values did not reach statistical significance. These findings suggest that changes in antioxidant enzyme activities may amplify the leishmanicidal effect in patients with CL. However, these changes may not only cause the killing of parasite but also may cause oxidative damage in the other cells or tissues. PMID- 14535170 TI - A second species of Tunga (Insecta, Siphonaptera) infecting man: Tunga trimamillata. PMID- 14535171 TI - [Development of seasonal malarial illness in a riparian region of Senegal]. PMID- 14535172 TI - Seventy-two cases of antidiabetic-induced hepatopathy treated by huatan yigan decoction. AB - In order to observe the therapeutic effects of Huatan Yigan Decoction ([symbol: see text] Decoction for resolving phlegm and replenishing the liver) on antidiabetic-induced hepatopathy, 129 cases were divided into two groups, 72 cases in the treatment group were administrated with Huatan Yigan Decoction and 57 cases in the control group with the liver-protecting drugs. Their liver function, plasma prothrombin time (PT), blood urea nitrogen and symptoms of the patients before and after treatment were observed during the three therapeutic courses. RESULTS: The total effective rate and cured rate of the treatment group manifested an obvious difference in comparison with those of the control group (P < 0.05 or P < 0.01). Even though the liver function, blood urea nitrogen (BUN) and symptoms in both groups were improved markedly, these indicators in the treatment group were improved more markedly than those in control group (P < 0.01). No adverse effect was found during the treatment. CONCLUSION: Huatan Yigan Decoction shows a repair action on hepatic injury. PMID- 14535173 TI - TCM treatment of Parkinson's syndrome--a report of 40 cases. PMID- 14535174 TI - TCM treatment of interstitial pneumonia with chronic cough--a case report. PMID- 14535175 TI - TCM treatment for 40 cases of rheumatoid arthritis with channel blockage due to yin deficiency. AB - To verify the therapeutic effects of the method of softening and lubricating the joints, and calming the endogenous wind in case of rheumatoid arthritis (RA) with the syndrome of channel blockage due to yin deficiency, 60 RA patients with the syndrome of channel blockage due to yin deficiency were randomly divided into a treatment group (40 cases) and a control group (20 cases) and treated respectively by the above method for the former and with Zheng Qing Feng Tong Ning Tablets ([symbol: see text]) for the latter. The result turned out to be that the effect in the treatment group was very satisfying. The treatment group obtained a better result in the accumulated points of syndrome and RA, morning rigidity of the joints, grip strength, 20m walking time and erythrocyte sedimentation rate (ESR) (P < 0.01 or P < 0.05). The above indicates that channel blockage due to yin deficiency is an important pathogenesis of RA, and calming the endogenous wind is a method of choice for treating RA. PMID- 14535176 TI - Personal experience in TCM treatment of reflux esophagitis. PMID- 14535177 TI - Differential treatment for membranoproliferative glomerulonephritis with TCM prescriptions plus triptoryph tablets--a report of 30 cases. PMID- 14535178 TI - Clinical application of bai du san. PMID- 14535179 TI - Clinical observation in 40 cases of postmenopausal coronary heart disease treated with yanghuo sanzi tang. PMID- 14535180 TI - Professor Ouyang Heng's experience in treating neurodermatitis. PMID- 14535181 TI - Clinical observation on treatment of acquired paralytic strabismus by acupuncture plus oral administration of Chinese herbs. PMID- 14535183 TI - Electro-acupuncture combined with plum-blossom needle tapping for treatment of supraorbital neuritis--a clinical observation of 59 cases. PMID- 14535182 TI - Clinical study on the needling and drug treatment of acute cerebral hemorrhage. PMID- 14535184 TI - Therapeutic effects of the point-injection therapy on primary trigeminal neuralgia--an observation of 103 cases. PMID- 14535185 TI - Piriformis syndrome treated by triple puncture with the bai hu yao tou maneuver. PMID- 14535186 TI - Acute lymphangitis treated by moxibustion with garlic in 118 cases. AB - TCM holds that acute lymphangitis is pathogenically caused by the accumulated internal fire and toxicity, and the external skin injury-induced infection, leading to toxicity going upward along the meridians. The treatment should be given to clear away heat and toxin. Blood letting by a three-edged needle may promote qi and blood circulation and remove the toxic heat. Garlic has the function of antagonizing Staphylococcus aureus, Bacillus dysenteriae and various kinds of bacteria. Since the volatile oil in moxa is antagonistic to bacteria, the burning moxa can also promote the local flow of qi and blood, and enhance the antagonistic effect of garlic on bacteria. Therefore, satisfactory curative results can be achieved by the therapy. PMID- 14535187 TI - Clinical observation in 46 cases of angina pectoris treated by the catgut embedding therapy. PMID- 14535188 TI - Point injection for treating nephritic colic in 101 cases. PMID- 14535189 TI - Two-hundred and ten cases of shoulder periarthritis treated by needling lingxia and sanjian. PMID- 14535190 TI - Thirty cases of acute lumbar sprain treated by acupuncture combined with point injection at tianzhu. PMID- 14535191 TI - Treatment of migraine with acupuncture at points pertaining to the liver and gallbladder channels. PMID- 14535192 TI - Clinical observation in 30 cases of chloasma treated by auricular point pressing and pricking. PMID- 14535193 TI - Treatment of hyperlipemia by acupoint catgut-embedding in 34 cases. PMID- 14535194 TI - Clinical experience in multiple needling therapy. PMID- 14535195 TI - Acupuncture and the elevation manipulation of massage for treatment of frozen shoulder. PMID- 14535196 TI - A follow-up study of 69 discharged SARS patients. AB - Sixty-nine patients with severe acute respiratory syndrome (SARS) discharged from Guangdong Provincial TCM Hospital were followed up from January to April 2003 during which the patients were asked to fill the questionnaire form and at the same time received blood routine examination, hepatic, renal, pulmonary and immune function tests, and spiral computerized tomography (CT) of the chest, color B-ultrasonography of the heart with the collected data treated by descriptive analysis and deductive analysis. The results showed that in the 69 followed-up patients, impairment of the hepatic function was found in 5 cases, hypoimmune state in 18, impediment of ventilation in the distal air passages with normal major air passages in 15, increased residual volume in 40, mild disturbance of pulmonary diffusion function in 14, incomplete absorption of inflammatory exudates, focal or multiple interstitial lesions, pulmonary interstitial fibrosis and pleural adhesion in 24; increased resistance or mild systolic hypertension in the pulmonary circulation, and segmental ischemia of the left myocardium in 34; and decreased visual acuity in 2. According to TCM differentiation 24 cases belonged to the type of deficiency of both qi and yin, 8 deficiency of both the heart and spleen, 37 depression of the liver and deficiency of the spleen, 18 intermingling with damp-heat, and 7 intermingling with stagnant blood. Some patients still had psychological problems. The study indicates that though clinically cured and discharged from hospital, some SARS patients have functional impairment of the heart, lung and liver, hypoimmune state as well as psychological problems, and need to be treated accordingly for a complete recovery. A rationale for suggested TCM treatment is expounded. PMID- 14535197 TI - The therapeutic effects of zhaohai (KI 6) on hoarseness in 92 cases. PMID- 14535198 TI - Effects of san qi on gastric secretion and protective factors of gastric mucosa in the rat with precancerous lesion of stomach. AB - In the model rat with precancerous lesion of stomach induced by the combined method of insertion of a spring into the pylorus and high salt hot paste, effects of San Qi ([symbol: see text] Radix Notoginseng) on gastric secretion and protective factors of stomach were investigated. The results indicated that gastric secretion, gastric mucosal blood flow (GMBF) and aminohexose content lowered significantly, and malondialdehyde (MDA) increased significantly (P < 0.01) in the model group as compared with the normal group; after treatment with San Qi Powder for 12 weeks, both gastric secresion and GMBF increased, and MDA content decreased as compared with the negative control group (P < 0.01), with no significant increase of aminohexose content. It is suggested that San Qi ([symbol: see text] Radix Notoginseng) may improve gastric secretion, and that increase of GMBF and antagonism against the lesion of oxygen free radicals are possibly one of its mechanisms. PMID- 14535199 TI - Effects of "moxibustion serum" on proliferation and phenotypes of tumor infiltrating lymphocytes. AB - Tumor infiltrating lymphocytes (TIL) were cultured with "moxibustion serum" (MS), and the results were examined by flow cytometry. The results indicated that MS could enhance the proliferation of TIL, accelerate it to reach the exponential growth phase, and assist recombinant interleukin 2 (rIL-2) to enhance successively the percentage of CD3+ positive cells, maintain the number of CD4+ positive T cells, promote greatly the percentage of CD8+ positive T cells among TILs, and reverse the CD4+/CD8+ ratio. Such cooperative effects rely on relative specificity of acupoints. It is suggested that MS is beneficial to the growth of TIL both in the aspects of proliferation and phenotypes. PMID- 14535200 TI - A survey on TCM treatment of polymyositis and dermatomyositis. PMID- 14535201 TI - Can TCM treatment be performed on the basis of differential diagnosis of the diseases instead of the syndromes? PMID- 14535202 TI - Essentials for the acupuncture treatment of radicular sciatica. PMID- 14535203 TI - [Are enemies' enemies friends?--plant-carnivore interactions mediated by herbivore-induced plant volatiles]. PMID- 14535204 TI - [Complex tritrophic interaction networks in cabbage field]. PMID- 14535205 TI - [Herbivore elicitors and plant signaling pathways involved in the production of herbivore-induced plant volatiles]. PMID- 14535206 TI - [Regulatory mechanism of plant volatile formation involved in chemical communications]. PMID- 14535207 TI - [Molecular basis of plant-plant communication mediated by plant volatiles]. PMID- 14535209 TI - [Vesicular glutamate transporter and glutamatergic signaling]. PMID- 14535208 TI - [Systemic regulatory system sustaining symbiosis and organ development in legume]. PMID- 14535210 TI - [Photobiological hydrogen production by cyanobacteria: toward development of renewable energy source alternative to fossil fuels]. PMID- 14535211 TI - Endotracheal tubes. AB - A variety of different endotracheal tubes are available for distinct purposes, though the majority of patients will be well served with the standard single lumen endotracheal tube. Specialized endotracheal tubes have been developed to aid in specific situations and novel tubes continue to be evaluated as clinicians strive for improved outcomes in various clinical conditions. Ultimately, the choice of an endotracheal tube depends on the purpose it is intended to serve. PMID- 14535212 TI - Tracheotomy application and timing. AB - Tracheotomy is one of the most commonly performed surgical procedures among critically ill patients. In the past, tracheotomy was delayed as long as possible in ventilator-dependent patients because of concerns regarding injury to the airway from the surgical procedure. Greater recognition of the benefits of tracheotomy in terms of greater patient comfort and mobility has promoted its earlier performance. No data identify an ideal time for tracheotomy. The decision to convert a patient from translaryngeal intubation to a tracheostomy requires anticipation of the duration of expected mechanical ventilation and the weighing of the expected benefits and risks of the procedure. The convenience of percutaneous tracheotomy performed in the ICU by critical care specialists without formal surgical training has further promoted the adoption of tracheotomy for ventilator-dependent patients. Regardless of the method for performing tracheotomy, meticulous surgical technique and careful postoperative management are necessary to maintain the excellent safety record of tracheotomy for critically ill patients. PMID- 14535213 TI - Percutaneous dilational tracheostomy techniques. AB - In summary, PDT is a bedside procedure that can be performed with very low morbidity by skilled practitioners. Established methods are undergoing constant evolution and we hope that technical adjuncts are improving patient safety. Evaluation of procedural modifications will require evaluation in randomized clinical trials. PMID- 14535214 TI - Percutaneous tracheostomy--special considerations. AB - Percutaneous tracheostomy is safe and highly effective in well-trained hands in establishing a long-term artificial airway. Most alleged contraindications and some suggestions on how the procedures should be performed likely stem from early trials when only "perfect candidates" were chosen. Most of those contraindications should not be viewed as prohibitions, but as suggestions related to the skill level and training of the operator. We have used this technique in many situations where the small incision and tamponading effect of the tracheostomy tube has been quite beneficial, in selected patients with coagulapathies and severe venous congestion from superior cava syndromes as well as thyroid cancers, and in whom operative approaches would have been difficult. Knowing one's level of expertise and comfort in choosing and rejecting patients and procedures accordingly is the key to keeping PT a procedure with an excellent safety record. As the experience with PT grows, more and more perceived contraindications will disappear. Studies will address the role of PT in children and as a means of establishing emergent airway access. Also, the exact coagulation limits will need to be established. Few contraindications will most likely remain absolute, such as active infections over the proposed entry site, uncontrollable bleeding disorders and excessive ventilatory and oxygenation requirements. In our institution, taking into account these absolute contraindications, fewer than 5% of patients in need of a tracheostomy in the intensive care unit will undergo a primary open procedure. PMID- 14535215 TI - Techniques of surgical tracheostomy. AB - Tracheostomy has become one of the most commonly performed procedures in the critically ill patient. Variations in technique and expertise have led to a wide range of reported procedural related morbidity and rarely mortality. The lack of prospective, controlled trials, physician bias and patient comorbidities further confound the decisions regarding the timing of tracheostomy. With careful attention to anatomy and technique, the operative complication rate should be less than 1%. In such a setting, the risk-benefit ratio of prolonged translaryngeal intubation versus tracheostomy begins to weight heavily in favor of surgical tracheostomy. At exactly what point this occurs remains undefined, but certainly by the tenth day of intubation, if extubation is not imminent, arrangements should be made for surgical tracheostomy by a team experienced with the chosen technique. PMID- 14535216 TI - Comparison of surgical and percutaneous dilational tracheostomy. AB - When significant clinical end points are considered, PDT is a cost-effective and safe alternative to ST in critically ill patients in the ICU when performed by skilled and experienced practitioners [1, 40]. There are insufficient data to establish clear superiority of either technique. Important advantages of PDT may include eliminating the need for operating room facilities and personnel by the performance of the procedure at the bedside and significantly decreasing the time interval between the decision to perform tracheostomy and the actual procedure [1, 2, 20]. PMID- 14535217 TI - Minitracheostomy. AB - Minitracheostomy is a safe bedside procedure that is a useful adjunct to facilitate secretion clearance in patients with sputum retention. It can also be used in a prophylactic fashion after operations in high-risk patients. Insertion requires a trained physician and an assistant. Complications are rare and are usually avoidable. PMID- 14535218 TI - The Montgomery T-tube tracheal stent. AB - The Montgomery T-tube is a valuable tracheal stent that provides a functional airway while supporting the tracheal mucosa. It is used in benign and malignant tracheal diseases and provides symptomatic relief to the majority of the patients. T-tubes are simple to insert and rarely cause serious complications. The use of T-tubes continues to gain popularity with the increasing incidence of benign trachea stenosis following the use of artificial airways. Physicians dealing with diseases of the airways should be familiar with the indications, contraindication, complications, and care of the Montgomery T-tubes. PMID- 14535219 TI - Acute complications of artificial airways. AB - In conclusion, though there has been a dramatic reduction in the acute complications of artificial airways in the last hundred years, it remains crucial for the intensivist/anesthesiologist to have an implicit understanding of the anatomy and physiology of the process of ETI. As new techniques such as PDT are introduced, we must investigate their utility compared with the current standard of care in the most rigorous fashion. Additionally, as many of the complications of ETI can lead to increases in morbidity and mortality, prompt diagnosis and management are essential. PMID- 14535220 TI - Long-term complications of artificial airways. AB - In summary, long-term complications of artificial airways are rare but important sequelae of artificial airways. Many of the potential long-term complications of translaryngeal intubation and tracheotomy are similar and overlapping. Although most patients who undergo these procedures tend to tolerate them without difficulties, significant morbidity and mortality may occur. Identifying the exact cause of the complication may not be possible at times, due to the multiple risk factors involved in the pathogenesis. It is hoped that understanding these potential complications will lead to a more vigilant preventive measures during the institution of long-term artificial airways and a judicious early search for the underlying pathology when a complication is suspected. PMID- 14535221 TI - Long-term care of the tracheostomy patient. AB - Care of the long-term tracheostomy patient is changing. By moving the initiation of tracheostomy out of the operating room and shifting responsibility for the procedure to the medical specialist, more patients are undergoing tracheostomy for a wider spectrum of diagnoses. With much of the aftercare now directed by the medical specialist, successful reintegration of the long-term tracheostomy patient into a productive life is dependent upon the collaborative care of several disciplines directed by the specialist. To effectively care for these challenging patients, it is critical for the physician who performs tracheostomy to be aware of the new caregiving role that is now theirs. PMID- 14535222 TI - Transtracheal oxygen catheters. AB - Over the past 20 years a variety of transtracheal catheters have been developed for long-term oxygen therapy. A modified Seldinger technique has been the standard in the past, but a more recent procedure for surgical creation of the tracheocutaneous tract presents a number of potential advantages. TTO should be administered as a program of care, and recent advances with a streamlined and shortened program have simplified and improved the delivery of a technology that has a number of potential benefits and established safety. TTO may further increase the oxygen conservation efficiency of demand oxygen controller devices, and studies have shown TTO to be a potential alternative to nasal oxygen, continuous positive airway pressure, and tracheotomy for severe obstructive sleep apnea. Very high flows (> 10 L/minute) of a humidified air/oxygen blend, termed transtracheal augmented ventilation, extend the physiologic benefits of TTO and have promise in both the outpatient nocturnal ventilatory support of patients with severe respiratory disease and in liberation of patients from prolonged mechanical ventilation. PMID- 14535223 TI - Management of hypoxemia during flexible bronchoscopy. AB - Under controlled conditions, FB is a safe procedure that has few significant adverse events. Significant hypoxemia may sometimes occur during FB despite the use of supplemental oxygen. UAO has been shown to be the dominant cause of hypoxemia during FB, and this is successfully managed with nasopharyngeal tube insertion. Other strategies that may need to be implemented include oxygen supplementation with intratracheal catheter, administration of sedation reversal medication, removal of the bronchoscope, bag-and-mask ventilation, and, rarely, endotracheal intubation and ventilation. Access to an anesthetist, availability of propofol, backup rigid bronchoscopy, and fluoroscopy are optional but desirable components in the bronchoscopy suite. PMID- 14535225 TI - Update on national certification for dialysis technologists. AB - Certification is an important step in the dialysis technologist's professional practice. We hope that candidates who are eligible will consider writing the national dialysis technology certification exam in order to validate the extensive and specialized body of knowledge required to practise. PMID- 14535226 TI - Over-the-counter remedies in chronic renal insufficiency: risks versus benefits. AB - It is common for individuals with renal insufficiency of varying degrees to supplement their prescriptive treatment with non-prescription medications, herbal medicines, nutritional supplements and vitamins. These products, although often perceived as "safe" to the consumer, do carry their own profiles of side effects and precautions. Renal insufficiency may put individuals at an even higher risk for adverse events. The purpose of this review is to give health care professionals an increased appreciation for the use of over-the-counter remedies by patients with renal insufficiency and the inherent risks associated with the use of these products. The uses and potential hazards of common non-prescription medicines, herbal medicines, nutritional supplements and vitamins will be discussed. PMID- 14535227 TI - The use of intradialytic parenteral nutrition to treat malnutrition: a case study. AB - Protein energy malnutrition in dialysis patients has been well-described in the literature. Most malnourished patients with end stage renal disease (ESRD) suffer from a mixed marasmus-kwashiorkor type of malnutrition with loss of both somatic and visceral protein mass. Malnutrition is associated with increased risk of morbidity and mortality. Up to 50% of patients on dialysis have protein energy malnutrition (Mortelmans & Vanholder, 1999). Malnutrition may be under-recognized and under-reported in dialysis patients. Malnutrition may result from inadequate food intake secondary to the uremic condition, nausea, vomiting, loss of appetite, altered taste and other physiologic conditions that impede food intake or metabolism. The usual indices of nutritional assessment--body weight, body mass index (BMI), anthropometrics, etc., may be inaccurate in patients with ESRD, as the results are often skewed by fluid retention. Therefore, we often rely on weight loss, bloodwork, a pre-dialysis low serum potassium, phosphorus and urea, as early signs of a decreased food intake. When patients are malnourished, measures such as oral supplements and/or tube feedings may be used to augment protein and calorie intake. However, when these interventions are inadequate to reverse the malnutrition condition, intradialytic parenteral nutrition (IDPN) should be implemented. Although there is no definite supportive data to show that the use of IDPN improves morbidity and mortality of dialysis patients, there are data to support that IDPN has positive effects on numerous nutritional parameters (Acchiardo, 2000; Capelli et al., 1994; Foulks, 1999; Hiroshige et al., 1998; Ikizler et al., 1995; Korzets et al., 1999; Mortelmans & Vanholder, 1999; Saunders et al., 1999; Smolle et al., 1995). In this article, we will discuss the causes of malnutrition in dialysis patients, the use of IDPN on one of our patients, and the potential complications associated with IDPN. PMID- 14535228 TI - Monthly hemodialysis nutrition lab report cards. PMID- 14535229 TI - The legal liabilities of nurse educators. PMID- 14535230 TI - When to cannulate new peripheral vascular accesses. PMID- 14535231 TI - The contribution of chemical speciation to internal dosimetry. AB - Speciation studies refer to the distribution of species in a particular sample or matrix. These studies are necessary to improve the description, understanding and prediction of trace element kinetics and toxicity. In the case of internal contamination with radionuclides, speciation studies could help to improve both the biokinetic and dosimetric models for radionuclides. There are different methods to approach the speciation of radionuclides in a biological system, depending on the degree of accuracy needed and the level of uncertainties accepted. Among them, computer modelling and experimental determination are complementary approaches. This paper describes what is known about speciation of actinides in blood, GI tract, liver and skeleton and of their consequences in terms of internal dosimetry. The conclusion is that such studies provide very valuable data and should be targeted in the future on some specific tissues and biomolecules. PMID- 14535233 TI - Scientific basis and development of internal dosimetry models (BIODOS). AB - Assessments of doses and risks to workers and members of the public exposed to radionuclides require biokinetic and dosimetric models which describe the behaviour of the radionuclides and provide tools for dose calculations. The overall objective of BIODOS is to improve the scientific basis of existing models and to provide new or improved models. One part aims to provide models targeting specific uncertainties, including estimates of doses and risks from radionuclides ingested by adults and children; estimates of the transfer of radionuclides to breast milk and doses to infants; development of systemic models to improve the interpretation of bioassays; and studies of the importance of heterogeneous distribution of dose within tissues and cells. The other part forms a comprehensive research programme on the inhalation of radionuclides and addresses four areas of uncertainty, namely ultrafine particles, inter-subject variation, absorption into blood and clearance mechanisms. PMID- 14535234 TI - Abstracts of the American Dietetic Association's Food and Nutrition Conference and Expo (FNCE). San Antonio, Texas, United States, October 25-28, 2003. PMID- 14535232 TI - Efficacy of 3,4,3-LI(1,2-HOPO) for decorporation of Pu, Am and U from rats injected intramuscularly with high-fired particles of MOX. AB - This study aimed to assess the efficacy of 3,4,3-LI(1,2-HOPO) for reducing uranium, plutonium and americium in rats after intramuscular injection of (U Pu)O2 particles (MOX). Sixteen rats were contaminated by intramuscular injection of a 1 mg MOX suspension and then treated daily for 7 d with LIHOPO (30 or 200 micromol kg(-1)) or DTPA (30 micromol kg(-1)). LIHOPO was inefficient for removing Pu, Am and U from the wound site. However, it reduced Pu retention in carcass and liver by factors of 2 and 6 respectively, and Am retention in carcass and liver by factors of 10 and 30. In contrast, the effect of LIHOPO on U was to decrease the retention in kidneys by a factor of 75. These results confirm that LIHOPO is a good candidate for use after contamination with MOX, in combination with localised wound lavage or surgical treatment aimed at removing most of the contaminant at the wound site. PMID- 14535235 TI - Abstracts of the VII Symposium on Research in Head and Neck Cancer. 4-6 September 2003, Dusseldorf, Germany. PMID- 14535236 TI - American College of Emergency Physicians Research Forum. October 12-13, 2003, Boston, Massachusetts, USA. Abstracts. PMID- 14535237 TI - EUROTOX 2003. Abstracts of the 41st Congress of the European Societies of Toxicology. Florence, Italy, September 28-October 1, 2003. PMID- 14535238 TI - Abstracts of the American Society for Therapeutic Radiology and Oncology 45th annual meeting. Salt Lake City, Utah, USA, October 19-23, 2003. PMID- 14535240 TI - Unraveling Inflammation. Abstracts of the 36th annual meeting of the Society for Leukocyte Biology. October 2-5, 2003, Philadelphia, Pennsylvania, USA. PMID- 14535239 TI - Missouri State Medical Association 2003-2004 membership directory. PMID- 14535241 TI - [Ultraschall 2003. 27th Three-country meeting of the Swiss, German, and Austrian Societies for Ultrasound in Medicine. 11-13 September 2003, Bregenz. Abstracts]. PMID- 14535242 TI - Abstracts of the 18th Congress of the International Diabetes Federation. Paris, France, 24-29 August 2003. PMID- 14535243 TI - Ophthalmological Society of the West Indies 14th annual congress. July 9-12, 2003. Montego Bay, Jamaica, West Indies. Abstracts. PMID- 14535244 TI - Skin grafts 2: management of donor site wounds in the community. AB - Skin grafting is a surgical procedure used to quickly restore skin integrity in large wounds or those wounds which cannot be directly closed by suturing. The procedure of skin grafting necessitates the creation of a second wound; the donor site. Although often viewed as secondary importance by surgeons once skin has been harvested from the area, it is the donor site which frequently causes complications such as pain/discomfort and slow healing (Wilkinson, 1997). Because skin graft sites and donor sites are viewed as part of a specialist practice, their wound management is regarded as being 'something different'. However, the donor site is a partial-dermal thickness wound and should be seen as such, rather than a 'special' wound. This may help to lessen the anxiety felt by both patient and nurse in dealing with donor site wounds. PMID- 14535245 TI - Growth factors: the wound healing therapy of the future. AB - 'Growth factors' is an umbrella term used to describe a variety of chemical messengers found in a wide range of tissues, which have a number of functions including the regulation of cell and tissue growth and development. Several growth factors have been proposed as ideal agents to promote wound healing. This review discusses their functions and how they can be applied in the management of chronic wounds. PMID- 14535247 TI - The nursing management of enterocutaneous fistulae: a challenge for all. AB - This article focuses on the principles of managing patients with enterocutaneous fistulae. Successful management involves comprehensive assessment, planning, implementation and evaluation of care and involves all members of the multidisciplinary team in both primary and secondary care. The main challenge for the nursing team is effective containment of the effluent to prevent excoriation of the skin thereby ensuring patient comfort. The article includes suggestions for choice and application of suitable appliances which have been successfully used by the authors. Early involvement in of patient and carer in all aspects of management is recommended for successful discharge of patients into the community. PMID- 14535246 TI - Introducing the use of sterile maggots into a primary care trust: overcoming barriers. AB - Larval therapy has been available to patients in hospital where there has been a clinical need. Not all primary care trusts realize the clinical and cost effectiveness of this treatment, or if they do, are prepared to recognise funding to allow patients to receive this treatment in the privacy of their own home. Funding can be an issue as can instigating changes in practice, particularly when there are clinicians and managers who have an aversion to the therapy. PMID- 14535248 TI - Women with bipolar disorder at higher risk for alcoholism. PMID- 14535249 TI - Millions of Americans suffer from severe depression. PMID- 14535250 TI - Studies explore effects of depression on pregnancy and beyond. PMID- 14535251 TI - Medical defence and insurance. 1952. PMID- 14535252 TI - The millennium bug: health and medicine around the year 1000. AB - This introductory paper has two main themes: first, what can be said about the diseases characteristic of the period in question; second, what generalizations are possible about the medicine that could be deployed against them. After a preliminary discussion of the extent to which the system of dating from Christ's incarnation governed the chronology of those who fall within our scope, the paper raises the question of whether the year 1000 prompted millenial 'fever'. This leads into an account of the concept of 'pathocoenosis', a tableau of the interrelated diseases prevalent at any one place and time. The limitations of the tenth- to eleventh-century evidence available for such a project are examined, philosophical objections to disease history reviewed, and agenda for future research outlined. The paper then turns to the main features of the medical texts on which any investigation of therapeutic practice in the early Middle Ages must depend: features such as amorphousness, mutability, lack of theory, elusive connection with clinical reality. Finally, some wider implications of the special issue are delineated-concerning presumed contrasts between East and West, and, with respect to the relationship of theory to practice, between early and later Middle Ages. PMID- 14535253 TI - The practice of medicine in England about the year 1000. AB - The paper begins with the medical practitioners of late Anglo-Saxon England, were apparently both physicians and surgeons, describing the kinds of ailments they are evidenced as treating. The majority were monastic; whether there were also lay medics is uncertain. Most Anglo-Saxon ecclesiastical foundations appear to have had an infirmary, where sick monks or nuns, or those who were about to die, were looked after. Some infirmaries may have tended lay people, but there do not appear to have been any hospitals in the later medieval sense. The rest of the paper looks at the contents of one manuscript written about the year 1000: London, British Library MS. Harley 585, which contains texts of three compilations in old English: the Herbarium and the Medicina de Quadrupedibus, both translated from Latin, and the Lacnunga, which is a collection of remedies from diverse sources, some translated from Latin, some of native origin, some wholly rational, some containing Christian or folkloric incantations and rituals, and including four metrical charms. Because of its lack of selectivity and orgarization, it gives an invaluable insight into the condition of, and the attitude to, Anglo-Saxon medicine about the end of the first millennium. PMID- 14535254 TI - Dr Monk's medical digest. AB - The Liber passionalis is an early and hitherto mostly unexplored example of a composite medical work on diagnosis and therapy, similar to the better-known compilations circulating under the titles Petroncellus and Gariopontus (Passionarius Galieni). It shows the efforts made to provide comprehensive coverage of morbid conditions drawn from a choice of the best sources available to the compiler, sources which in some instances complement or enhance our knowledge of ancient medicine in a way overlooked by specialists in the field for a long time and which provide the best clue when trying to assess medical expertise at the turn of the first millennium. The paper explores the transmission, structure and sources of the Liber passionalis, touching on the body of medical literature available in the early Middle Ages. PMID- 14535255 TI - Medicine and hagiography in Italy c. 800-c. 1000. AB - A vast range of sources, from chronicles, hagiography and the liturgy to medical manuscripts and charters, is at our disposal for the study of health and healing in Italy between the ninth and eleventh centuries. What is needed in order to exploit this material is a methodology for the careful examination of sources in their regional, social and political context. I focus on what hagiography can contribute to the study of medicine and disease, discussing in detail two ninth century episcopal hagiographies, the Milanese second Vita Ambrosii, and the Vita Barbati from Benevento in southern Italy. At the same time, however, I stress the limitations of studying texts in isolation, given that one region, city, or community could be simultaneously producing a number of different kinds of evidence, with possibly more than one view of sickness and healing. There is great potential for medical history in further investigation of the archaeological, liturgical, and charter evidence from early medieval Italy. The study of all surviving manuscripts from this period, not just medical words, can provide a bridge between the detailed examination of a particular text and a discussion of the wider literary and cultural traditions into which they fitted. PMID- 14535256 TI - Signs and senses: diagnosis and prognosis in early medieval pulse and urine texts. AB - The character of early medieval medical manuscripts makes it difficult to generalize about the nature of medical knowledge in this period. In order to reconstitute one field of medical science, namely diagnosis and prognosis, while avoiding the pitfalls of unjustified generalization, this essay limits itself to reconstructing the understanding of pulse and urine inspection available in a particular place and time: the Italian monastery of Monte Cassino at the end of the first millennium. The available texts reveal little about the rationale behind these bedside techniques; indeed, pulse and urine seem to be signs without any semiotics, any underlying theory. The clue to this paradox is the fact that these texts see pulse and urine as primarily prognostic rather than diagnostic. Prognosis was understood to be analogous to forms of intuition, judgement, revelation, and prophecy that operated outside the logic of causality. Hence a fully rationalized semiotics was not regarded as necessary for effective medical practice. PMID- 14535257 TI - Medical practice and manuscripts in Byzantium. AB - Scholars, past and present, have belittled Byzantine medicine for its perceived static and derivative nature. Applied to the medicine of the centuries immediately before and after the year 1000, these criticisms, though apparently sustainable, fail to recognize its underlying vigour. It was a practical craft medicine, and one which used elements of magic and religion to compensate for the intractability of the many diseases that were not amenable to medicine. These centuries were a time when hospitals flourished in Byzantium, and this paper assembles and describes some of the manuscripts that can be associated with them. The influence of islamic medicine, which had hitherto borrowed from Byzantium, is also examined. If the Byzantine medicine of this period could not claim great originality or innovation, it had none the less distilled what was best and most useful from the long and often complex medical writings of antiquity, ensured its transmission, and preserved much from earlier times that would otherwise now be lost. PMID- 14535258 TI - Practice versus theory: tenth-century case histories from the Islamic Middle East. AB - Medicine and disease in medieval Islam have thus far been approached through theoretic medical treatises, on the assumption that learned medical texts are a transparent account of reality. A question yet to be sufficiently explored is the extent to which the ideas and theoretical principles they contain were actually carried out in practice. This paper deals with the description of diseases occurring in a tenth-century Casebook (Kitab al-Tajarib) by Abu Bakr Muhammad ibn Zakariya' al-Razi (known to Europeans as Rhazes)-the largest and oldest collection of case histories, so far as is known, in medieval Islamic medical literature. Since the author was a prolific medical writer, this study also includes a review of his medical and therapeutic principles dealing with eye diseases, as described in his learned treatises, and a comparison with those therapies actually employed in his everyday practice, as exemplified by the Casebook. The comparative analysis shows that the medical knowledge and the therapeutic advice so meticulously described in theoretical works were not paralleled in the physician's medical performance. On the contrary, it appears that learned treatises served other purposes than determining medical practice. PMID- 14535259 TI - The practice of surgery in Islamic lands: myth and reality. AB - This paper analyses evidence for the practice of surgery, as opposed to its theory, in the Islamic Middle East at the end of the first millennium. The inclusion in formal Arabic medical treatises of complex or invasive surgical procedures is compared with the lack of evidence for their actual performance, as well as with statements to the effect that such techniques were unknown at that time or should be avoided. Areas in which there is greater evidence of the practice of surgery-such as the removal of superficial growths and the treatment of eye diseases-are also discussed. In particular, the paper focuses upon treatises by 'Abu al-Qasim al-Zahrawi (known to the Europeans as Albucasus), Abu Bakr Muhammad ibn Zakariya' al-Razi (Rhazes), 'Ali ibn al-'Abbas al-Majusi (Haly Abbas) and Ibn Sina (Avicenna). PMID- 14535260 TI - High-dose chemotherapy for breast cancer. PMID- 14535261 TI - High-dose chemotherapy for breast cancer. PMID- 14535262 TI - High-dose chemotherapy for breast cancer. PMID- 14535263 TI - Nitrous oxide and 5,10-methylenetetrahydrofolate reductase. PMID- 14535264 TI - Stem cells. PMID- 14535265 TI - Implementation of the Methamphetamine Anti-Proliferation Act; thresholds for retailers and for distributors required to submit mail order reports; changes to mail order reporting requirements. Final rule. AB - This regulation implements the new threshold requirements and mail order reporting requirements of the Methamphetamine Anti-Proliferation Act of 2000 (MAPA), which was enacted on October 17, 2000. DEA is amending its regulations to reduce the thresholds for pseudoephedrine and phenylpropanolamine for retail distributors and for distributors required to submit mail order reports. Also, DEA is amending its regulations to require mail order reports for certain export transactions. DEA is codifying exemptions from the mail order reporting requirements for certain distributions to nonregulated persons and certain export transactions. This rule is consistent with the intent of MAPA to prevent the diversion of drug products to the clandestine manufacture of methamphetamine and amphetamine, and simultaneously reduce the industry reporting burden. PMID- 14535266 TI - Case 11-2003: ulcerative colitis and primary sclerosing cholangitis in a 14-year old boy. PMID- 14535267 TI - Pathology, danger, and power: women's and physicians' views of pregnancy and childbirth in Weimar Germany. AB - This article shows how the very real risks of reproduction were culturally constituted by expectant mothers and physicians in Germany following the first World War. Both saw a strong potential for maternal and fetal death and injury. However, expectant mothers and physicians differed in the extent to which they regarded risk as contingent or inherent. Most physicians believed that pregnancy and childbirth first became dangerous in conjunction with a wide variety of other ailments, including complications specific to pregnancy. This view of pathology held a promise of prevention or at least treatment. Physicians further participated in inventing new, modern understandings of pathology, which also encompassed factory work as a reproductive hazard. Expectant mothers, in contrast, viewed pregnancy and childbirth as inherently dangerous based on their experiences and those of the women they knew, which not infrequently involved illness, disability, or close encounters with death. Many of them also held the ancient idea that a woman could harm the foetus with her imagination. As a result, though women varied in their beliefs (influenced by class, education, region, and religion), as a group they feared childbirth. Paradoxically, competition among lay and medical ideas gave women new options while also gradually enhancing medical authority. PMID- 14535268 TI - Patients' rights and the law of contract in eighteenth-century England. AB - The study of 34 lawsuits between practitioners and patients shows how the law relating to contracts was brought to bear on convicts over medical practice in eighteenth-century England. It shows that patients in this period had rights, and explores them though the practice of the courts. The article illuminates two substantial changes: the decline of the contract of cure, and the creation of the patient's right to disregard bills for fees from physicians and apothecaries. It argues that the common law created a medical market-place that was differentiated by the legal status of the patient, and that this affected the character of many healing relationships, even though the legal scrutiny of a patient-practitioner encounter was a relatively infrequent occurrence. The inability of married women and minors to make contracts, along with the legal and customary responsibility of employers for the health of servants and apprentices, meant that many patients in eighteenth-century England were not the autonomous consumers of medical care that existing histories of the patient in this period suggest. The essay thus investigates the impact of the law on the culture of medicine, whilst using legal sources to address questions about power in healing relationships. PMID- 14535269 TI - Slavery, smallpox, and revolution: 1792 in Ile de France (Mauritius). AB - In 1792 a slave-ship arrived on the french Indian Ocean island of Ile de France (Mauritius) from South India, bringing with it smallpox. As the epidemic spread, a heated debate ensued over the practice of inoculation. The island was in the throes of revolutionary politics and the community of French colonists were acutely aware of their new rights as 'citizens'. In the course of the smallpox epidemic, many of the political tenisons of the period came to focus on the question of inoculation, and were played out on the bodies of slaves. Whilst some citizens asserted their right, as property owners, to inoculate their slaves, others, equally vehemently, objected to the practice and asserted their right to protect their slaves from infection. Eighteenth-century colonial medicine was largely geared to keeping the bodies of slaves and workers productive and useful, but formal medicine never had a monopoly. Slaves on Ile de France brought with them a rich array of medical beliefs and practices from Africa, India, and Madagascar. We have little direct historical evidence for these, but we do know that many slaves came from areas in which forces of smallpox inoculation were known and practised. PMID- 14535270 TI - Trauma and bereavement: conceptual and clinical issues revolving around relationships. AB - The article re-examines trauma and bereavement and the category of loss considered to be traumatic bereavement. It is argued that the perspective of the relation to the deceased is an insufficiently acknowledged source of traumatic disruption following bereavement. The significance of the relational aspect in bereavement is present in the paradigm of the Two-Track Model of Bereavement (S. Rubin, 1981, 1999). The implications of this approach are examined in the case of a soldier exposed to trauma and death. The case is examined from a traditional trauma perspective stressing exposure to life threat and again from a perspective stressing the significance of the relationship to the deceased. An examination of the different intervention strategies and their outcome lends support to the importance of the relational aspects in this case of trauma and supports the use of the Two-Track Model of Bereavement. The location of trauma as basic to the experience of all bereavement for its impact on the reorganization vis a vis the representation of the loved one now deceased is considered as fundamental to the trauma of all bereavement. The additional contribution of bereavement occurring under conditions of life threat (traumatic situations) is an important feature potentially impacting the bereavement process, but it is not what makes bereavement traumatic per se. An additional source of trauma in bereavement, occurring when the previous representation or introject of the deceased is shattered, is considered for its status as an additional source of complication in the traumas of bereavement. PMID- 14535271 TI - Physicians and famine in Japan: Takano Choei in 1830s. AB - Takano Choei (1804-50) was a rangakusha: a scholar of Dutch learning, and a physician of Western medicine. Although he was one of the foremost scholars in his field, his career was cut off in its prime when he became the victim of an intrigue in 1839. As a result, much of his early work has been overshadowed by the political events that followed. This article takes a different approach to the writing of Choei's history by presenting an example of his work as a physician in the period before his arrest. It examines the responses of Choei and his colleagues to the famine and pestilence which ravaged the population during the Tempo Famine of the 1830s. The study is based on two short articles: one concerns the precautions people should take against epidemic disease, while the other deals with hardy, rapidly maturing crops thought helpful in warding off starvation. The focus of this article is on the way knowledge from Western sources was 'received'; that is, how it was actively assimilated and transformed, through a process of cultural exchange. PMID- 14535272 TI - Taking prisoners: Havelock Ellis, Sigmund Freud, and the construction of homosexuality, 1897-1951. AB - This paper addresses the efforts of both Havelock Ellis and Sigmund Freud to posit a theory of homosexuality, and especially considers their efforts to (re )negotiate each other's theories. Its central premise derives from the sociology of scientific knowledge: that it is not what is written, but the way that what is written is treated by ensuing experts, that makes knowledge. In the case study used in this paper, Ellis and Freud struggle to posit what they consider to be the proper model for understanding homosexual desire. They utilize aspects of each other's word, but are careful not to appear to be following each other too closely. Such a struggle to establish different schools of thought is exemplified by the informal negotiations engaged in when a student, Joseph Wortis, made contact with both Freud and Ellis. Again following sociology of scientific knowledge precepts, these informal negotiations (contained in published and archival letters) are used to show how knowledge claims are constructed, deconstructed any reconstructed by the actors who have stakes in the outcome of what is to be regarded as knowledge in the relevant communities. PMID- 14535273 TI - Framing tropical disease in London: Patrick Manson, Filaria perstans, and the Uganda sleeping sickness epidemic, 1891-1902. AB - Much of the historical literature on tropical medicine represents the periphery as the chief site for the production of western knowledge about disease in the British empire. This study on the Filaria perstans-sleeping sickness hypothesis revises this perspective by showing how the imperial metropole functioned as a culture space for the construction of knowledge about the empire. Beginning in 1891, Patrick Manson used the publicity resources of London to generate a rhetorical imperative for the confirmation of his hypothesis without ever leaving Britain. Later, while he was medical adviser to the imperial state, the 1900 sleeping sickness epidemic in Uganda presented Manson with a unique opportunity to determine the validity of his hypothesis. By exaggerating the possible spread of the epidemic privately among Foreign Office personnel and publicly in the medical press, he succeeded in mobilizing the first Royal Society sleeping sickness research expedition to Africa in 1902. While this expedition ultimately disproved Manson's hypothesis, this outcome ironically created the very conditions for the identification of the actual causal agent (Trypanosoma gambiense) and its vector (tsetse fly) by Aldo Castellani and David Bruce respectively. PMID- 14535274 TI - Social history of medicine in Spain: points of departure and directions for research. AB - The development and extension of activities in the domain of die social history of medicine in Spain since the 1960s is reviewed. Attention is paid, first, to the institutional setting and theoretical background of this broad line of research, taking the perspective of the professionalization of the subject of the 'history of medicine' as an under-graduate discipline in the syllabus of the Faculties of Medicine. Secondly, four main directions for research are outlined, namely disease and society, health policies, health professions and gender and medicine, including critical attention to the principal works, active authors, and working and narrative styles. PMID- 14535275 TI - Sources in the history of occupational health: the Turner & Newall archive. AB - Sources in the History of occupational health are scanty-a reflection perhaps of the contentious nature of the documentation. In the UK, asbestos company records have until recently been unavailable and consequently historians and policy makers have been hindered from exploring a major public health issue. In 1995, however, Chase Manhattan Bank in New York sued the leading British asbestos producer, Turner & Newall, in a property-damage claim. During pre-trial discovery, Chase had copied a large proportion of Turner & Newall's vast archive and under American law was able to put the records in the public domain. This article describes how this collection of records-perhaps the largest anywhere on the history of an occupational health hazard-was generated. It also suggests ways of navigating the documents; discusses the nature of the material; and the archive's uses to medical and industrial historians. PMID- 14535276 TI - Social construction in a cold climate: a response to David Harley, 'Rhetoric and the social construction of sickness and healing' and to Paolo Palladino's comment on Harley. PMID- 14535277 TI - Psychiatric treatment in the twentieth century. PMID- 14535278 TI - Visions of the dead: imagination and mourning. AB - Contemporary research recognizes the existence of an ongoing attachment between the mourner and the deceased. This research tends to focus on the subjective aspects of the attachment--the mourner's memory and emotions. A complementary perspective is offered by archetypal psychology, which considers the deceased to exist outside the mourner's subjectivity as an autonomous image. This approach is illustrated by an exemplary case: a woman named Verda who experienced the death of her twin sister,Vera.Their account suggests that Verda's mourning derives from her encounters with Vera's continuing imaginal presence. PMID- 14535280 TI - Abstracts of the British Society for Haemostasis and Thrombosis annual meeting. 4 6 September 2002, University of Leicester. PMID- 14535279 TI - Managing bereavement in the classroom: a conspiracy of silence? AB - The ways in which teachers in British schools manage bereaved children are underreported. This article reports the impact of students' bereavement and their subsequent management in primary and secondary school classrooms in Southeast London. Thirteen school staff working in inner-city schools took part in in-depth interviews that focused on the impact of bereaved children on the school and how teachers responded to these children.All respondents had previously had contact with a local child bereavement service that aims to provide support, advice, and consultancy to children, their parents, and teachers. Interviews were audiotaped, transcribed verbatim, and analyzed using ATLAS-ti. Three main themes were identified from analysis of interview data. Firstly, British society, culture, local communities, and the family were significant influences in these teachers' involvement with bereaved students. Secondly, school staff managed bereaved students through contact with other adults and using practical classroom measures such as "time out" cards and contact books. Lastly, teachers felt they had to be strong, even when they were distressed. Surprise was expressed at the mature reaction of secondary school students to deaths of others. The article recommends that future research needs to concentrate on finding the most effective way of supporting routinely bereaved children, their families, and teachers. PMID- 14535281 TI - Strategies for reducing racial and ethnic disparities in health. AB - This report is the fourth in a series of programs made possible by a grant to Grantmakers In Health (GIH) from The Commonwealth Fund to bring grantmakers together with experts in policy, practice, and research to exchange information and ideas about key health issues facing the nation. On May 18, 2000, GIH convened a select group of grantmakers along with officials from the U.S. Department of Health and Human Services and other experts in research and practice for an Issue Dialogue on racial and ethnic disparities in health. Intended to both share lessons learned and stimulate new ideas, the Issue Dialogue proved to be a rich, although difficult discussion about the conditions that give rise to health disparities. Among the ideas discussed were the interaction of race and socioeconomic status on health, environmental justice, and the complexities of working with different racial and ethnic groups. At the same time, the forum focused on what health funders can do to support the development of programs to improve the health of minorities. This Issue Brief brings together key points from the day's discussion with factual information on racial and ethnic disparities in health and grantmaker activities drawn from a background paper prepared for Dialogue participants. PMID- 14535282 TI - Long-term care quality: facing the challenges of an aging population. AB - As part of its continuing mission to serve trustees and staff of health foundations and corporate giving programs, Grantmakers In Health (GIH) convened a select group of grantmakers and national experts who have made a major commitment to improve the quality of long-term care, particularly the care provided in nursing homes. The roundtable explored various factors influencing both the quality of care provided to elderly patients, as well as the quality of their lives. The discussion ultimately centered upon the importance of grantmaker involvement to improve the quality of long-term care, including the services delivered, training for professional and paraprofessional staff, continued research and evaluation, and public policies regulating the long-term care industry. This report brings together key points from the day's discussion with factual information on demographic, financing, and public policy trends drawn from a background paper prepared for the meeting. When available, recent findings, facts, and figures have been incorporated. PMID- 14535283 TI - Progress and peril: examining antibiotic resistance and systemic contaminants. AB - As part of its ongoing mission to inform and support trustees and staff of health foundations and corporate giving programs, Grantmakers In Health (GIH), in conjunction with the Health and Environmental Funders Network (HEFN), held a day long Issue Dialogue on two important environmental health issues: antibiotic resistance and systemic contaminants. Experts and advocates from many organizations came together to discuss concerns and opportunities, as well as current and potential roles for health philanthropy. This Issue Brief brings together key points from the day's discussion with factual information on environmental health drawn from a background paper prepared for the participants. PMID- 14535284 TI - Early childhood development: putting knowledge into action. AB - As part of its continuing mission to serve trustees and staff of health foundations and corporate giving programs, Grantmakers In Health (GIH) convened a select group of grantmakers and national experts who have made a major commitment to improve the health and well being of young children. The roundtable explored the latest research examining early childhood development, as well as public and private programs serving families with young children. The discussion ultimately centered upon the importance of grantmaker involvement to improve early childhood development, including the services delivered to young children and their families, training for professionals, and continued research and evaluation. This report brings together key points from the day's discussion with factual information on demographic, health and human services, and public policy trends drawn from a background paper prepared for the meeting. When available, recent findings, facts, and figures have been incorporated. PMID- 14535285 TI - Long-term remission of Crohn's disease treated with thalidomide: a seminal case report. AB - A 31-year-old female with severe Crohn's disease for 15 years who had been treated with corticosteroids and 6-mercaptopurine, was treated with thalidomide initially for erythema nodosum. While on thalidomide all symptoms of Crohn's disease disappeared and she was able to discontinue all other drugs. At this writing she has been on thalidomide as sole therapy for over 4 years with the exception of a 5-week hiatus, during which time her symptoms recurred, but again disappeared after resumption of thalidomide therapy. This case suggests that thalidomide may be a useful therapy for Crohn's disease and provides impetus for a clinical trial of thalidomide for Crohn's disease. PMID- 14535286 TI - The C-terminal half of heat shock protein 90 represents a second site for pharmacologic intervention in chaperone function. AB - The molecular chaperone heat shock protein 90 (Hsp90) is required for stability and function of multiple mutated, chimeric, and over-expressed signaling proteins that promote cancer cell growth and/or survival. It is also critical for the function of many normally expressed proteins, including protein kinases, steroid receptors and other transcription factors, and it may protect the cell from incapacitating or deleterious mutations. The recent identification of a nucleotide binding pocket within the first 220 amino acids of the protein, together with the discovery that at least two structurally distinct classes of antibiotic can replace nucleotide at this site and alter chaperone activity, has deservedly focused attention on Hsp90's aminoterminus as an important regulator of function. However, data continue to accumulate pointing to the C-terminal half of the chaperone as an equally important regulator of activity, and small molecules that bind to this portion of Hsp90 have been identified. PMID- 14535287 TI - Advancing quality through patient safety. PMID- 14535289 TI - Filling the gap: strategies for improving oral health. AB - As part of its continuing mission to serve trustees and staff of health foundations and corporate giving programs, Grantmakers In Health (GIH) convened, with the Children's Dental Health Project (CDHP), a select group of grantmakers and national experts who have made a major commitment to improving oral health. This Issue Dialogue - held on May 16,2001, in Washington, DC - explored current challenges related to oral health in the United States, and highlighted public and private sector initiatives to overcome these challenges. The roundtable also illustrated current activities and future opportunities for foundations in the area of oral health. This Issue Brief synthesizes key points from the day's discussion with a background paper previously prepared for Issue Dialogue participants. It includes quantitative and qualitative information on oral health as well as profiles of public sector, private sector, and grantmaker strategies for promoting improvements. PMID- 14535290 TI - Weighing in on obesity: America's growing health epidemic. AB - On October 31, 2001, Grantmakers In Health (GIH) convened an Issue Dialogue in Washington, DC, on the problem of obesity and its implications for personal and public health. The purpose of this meeting was to help the staff and trustees of health foundations and corporate giving programs understand the nature of today's epidemic and explore ways in which they could play a role in identifying and promoting effective solutions. In addition to discussing the scope and implications of overweight and obesity in the United States, presenters and discussants provided insights into the strategies that grantmakers might want to consider to address this serious health problem. This Issue Brief incorporates the information and ideas shared during the meeting into the background paper which was prepared for participants at the Issue Dialogue. It offers detailed data on the prevalence and growth of overweight and obesity in various populations, discusses their impact on health and the costs of care, reviews the major causes, and suggests strategies for both prevention and treatment. In the context of those strategies, this report also profiles a number of efforts by both grantmakers and government agencies to tackle some of the root causes of this important public health issue. PMID- 14535291 TI - Training the health workforce of tomorrow. AB - As part of its continuing mission to serve trustees and staff of health foundations and corporate giving programs, Grantmakers In Health (GIH) convened a small group of grantmakers and national experts concerned about health workforce issues. This roundtable - held on October 31, 2001, in Washington, DC - explored various issues related to the supply, composition, and competency of the health workforce, and the role that these factors play in maintaining and improving the health status of individual patients and broader populations. The session also highlighted the current activities of and future opportunities for foundations.This Issue Brief synthesizes key points from the day's discussion with a background paper prepared for roundtable participants. It includes quantitative and qualitative information on workforce issues and profiles public sector and grantmaker strategies for addressing workforce problems. PMID- 14535292 TI - Rx for progress: putting patient safety into practice. PMID- 14535293 TI - Examining e-health. AB - As part of its continuing mission to serve trustees and staff of health foundations and corporate giving programs, Grantmakers In Health (GIH) convened a small group of representatives from foundations, healthcare organizations, and the technology field to share their experiences and expertise on electronic health (e-health). This roundtable, held April 28, 2002, in Chicago, Illinois, highlighted emerging opportunities and challenges for foundations that wish to fund e-health initiatives. This report summarizes some of the key points from the day's discussion, providing both an introduction for grantmakers new to this area and offering the perspectives of some funders already experienced in funding e health activities. PMID- 14535294 TI - Positive youth development: a pathway to healthy teens. AB - As part of its continuing mission to serve trustees and staff of health foundations and corporate giving programs, Grantmakers In Health (GIH) convened a group of experts from the fields of philanthropy, research, government, and health care to examine recent research on youth development and effective approaches for promoting healthy decision making by adolescents. This Issue Dialogue, Positive Youth Development: A Pathway to Healthy Teens-held June 13, 2002 in Washington, DC-provided an opportunity for grantmakers to explore how they can apply youth development principles and asset-based strategies to their grantmaking to promote healthy development and healthy decision making by the adolescents in their communities. This Issue Brief incorporates the information and ideas shared during the meeting into a background paper prepared for participants at the Issue Dialogue. It provides an introduction to positive youth development; describes the characteristics of effective services and programs; reviews the research in this area; and provides examples of programs sponsored by the government, national organizations, and foundations that are intended to support and promote positive youth development. PMID- 14535295 TI - Turning the tide: preserving community mental health services. AB - As part of its continuing mission to serve trustees and staff of health foundations and corporate giving programs, Grantmakers In Health (GIH) convened a group of experts from the fields of philanthropy, research, government, and health care on September 19, 2002 to examine the status of community-based services for people with mental disorders. This Issue Dialogue, "Turning the Tide: Preserving Community Mental Health Services," explored how health grantmakers can support community programs that provide critical mental health intervention and treatment services to children and adults. This Issue Brief synthesizes key points from the day's discussion with a background paper previously prepared for Issue Dialogue participants. It includes quantitative and qualitative information on mental health, as well as profiles of public sector, private sector, and grantmaker strategies for promoting improvements. PMID- 14535296 TI - Strengthening the public health system for a healthier future. AB - On November 6, 2002, Grantmakers In Health (GIH) convened a group of experts from the fields of philanthropy, research, government, and policy to examine the nation's public health infrastructure and explore opportunities for grantmakers to strengthen and sustain this fragile and troubled system. During the course of this meeting, representatives from the Institute of Medicine, which recently released a report on this topic, and the Centers for Disease Control and Prevention (CDC) discussed the role of the national public health system and reviewed weaknesses in its infrastructure. Elaborating on these points, local public health leaders offered an on-the-ground perspective of the challenges they face as they struggle to maintain and improve the health of the public at the same time as they prepare for the possibility of bioterrorism. Finally, several foundations with experience in this area shared their strategies for helping to buttress the infrastructure so vital to the health of the nation. This Issue Brief builds on a paper prepared in advance of the meeting by incorporating the highlights from presentations and discussions that took place at GIH's Issue Dialogue. It also incorporates comments and discussion from GIH's 2002 Fall Forum intensive breakout session Worst Case Public Health Scenario: Can Philanthropy Respond? Following a brief introduction to the public health system and its evolution over the past several decades, the Issue Brief discusses essential elements of the public health infrastructure, reviews the issues confronting each of these elements, and profiles various philanthropic initiatives aimed at repairing and improving the public health infrastructure. It concludes with an in depth look at the infrastructure issues involved in emergency preparedness. PMID- 14535297 TI - In the right words: addressing language and culture in providing health care. AB - As part of its continuing mission to serve trustees, executives, and staff of health foundations and corporate giving programs, Grantmakers In Health (GIH) convened a group of experts from philanthropy, research, health care practice, and policy on April 4, 2003, to discuss the roles of language and culture in providing effective health care. During this Issue Dialogue, In the Right Words: Addressing Language and Culture in Providing Health Care, health grantmakers and experts from policy and practice participated in an open exchange of ideas and perspectives on language access and heard from fellow grantmakers who are funding innovative programs in this area. Together they explored ways to effectively support comprehensive language services, including the use of interpreters and translation of written materials. This Issue Brief synthesizes key points from the day's discussion with a background paper previously prepared for Issue Dialogue participants. It focuses on the challenges and opportunities involved with ensuring language access for the growing number of people who require it. Sections include: recent immigration trends and demographic changes; the effect of language barriers on health outcomes and health care processes; laws and policies regarding the provision of language services to patients, including an overview of public financing mechanisms; strategies for improving language access, including enhancing access in delivery settings, promoting advocacy and policy change, improving interpreter training, and advancing research; and roles for foundations in supporting improved language access, including examples of current activities. The Issue Dialogue focused mainly on activities and programs that ensure linguistic access to health care for all patients. Although language and culture are clearly inseparable, a full exploration of the field of cultural competence and initiatives that promote its application to the health care setting are beyond the scope of this Issue Brief. The day's discussion did, however, raise provocative issues of culture that are reflected throughout this report. PMID- 14535298 TI - The capacity of duckweed to treat wastewater: ecological considerations for a sound design. AB - Duckweed species are promising macrophytes for use in sustainable wastewater treatment due to their rapid growth, ease of harvest, and feed potential as a protein source. This paper reviews growth rates of different duckweed species on wastewater and ammonia toxicity to duckweed and summarizes insights into the mechanism of organic matter and nutrient removal. Results were gained from laboratory experiments in small, shallow, duckweed-covered semicontinuous batch systems. Growth rates on different types of wastewater vary considerably among different species. Ammonia is toxic for duckweed in both the ionized and un ionized forms. Duckweed, however, can be used to treat wastewater containing very high total ammonia concentrations as long as certain pH levels are not exceeded. The degradation of organic material is enhanced by duckweed through both additional oxygen supply and additional surface for bacterial growth. The duckweed mat with attached bacteria and algae is, independent of the loading rates, responsible for three-quarters of the total nitrogen (N) and phosphorus (P) loss in very shallow systems. Based on our results we suggest that full-scale pilot plants with duckweed should be shallower than the range encountered in the literature. A harvesting schedule that allows doubling times of 2 to 3.5 d, maintenance of a full coverage, and plug flow conditions are recommended. PMID- 14535299 TI - Potential environmental benefits of ionophores in ruminant diets. AB - A concern of the USEPA is the volatilization of NH3 from animal manure and CH4 produced from ruminal fermentation. Excess N in the environment has been associated with adverse effects on human health, and CH4 and N2O emissions are sources of greenhouse gases. The objectives of this paper are to summarize and quantify the benefits of ionophores, principally monensin, in decreasing NH3 and CH4 emissions to the environment and reducing resource utilization in cattle (Bos spp.) production. The data indicate that monensin in the diets of ruminants may decrease protein degradation in the rumen and may increase feed protein utilization by an average of 3.5 percentage units. These changes would have an effect in reducing N losses and decreasing fecal N and the amount of protein that must be fed to meet animal requirements. Additionally, CH4 is produced by enteric fermentation in ruminants, which is responsible for about 33 to 39% of CH4 emissions from agriculture. Ionophores can reduce CH4 production by 25% and decrease feed intake by 4% without affecting animal performance. The inclusion of monensin in beef and dairy cattle diets may benefit air quality by reducing CH4 and N emissions and water quality by reducing N in manure, which can potentially leave the farm through leaching into ground water and through runoff into surface water. PMID- 14535300 TI - Growth and yield responses of potato to mixtures of carbon dioxide and ozone. AB - Elevated carbon dioxide (CO2) concentrations in the atmosphere can stimulate plant growth and yield, whereas ground-level ozone (O3) concentrations cause the opposite effect in many areas of the world. Recent experiments show that elevated CO2 can protect some plants from O3 stress, but this has not been tested for most crop species. Our objective was to determine if elevated CO2 protects Irish potato (Solanum tuberosum L.) from foliar injury and suppression of growth and yield caused by O3. An O3-resistant cultivar (Superior) and an O3-sensitive cultivar (Dark Red Norland) were exposed from within 10 d after emergence to maturity to mixtures of three CO2 and three O3 treatments in open-top field chambers. The three CO2 treatments were ambient (370 microL L(-1)) and two treatments with CO2 added to ambient CO2 for 24 h d(-1) (540 and 715 microL L( 1)). The O3 treatments were charcoal-filtered air (15 nL L(-1)), nonfiltered air (45 nL L(-1)), and nonfiltered air with O3 added for 12 h d(-1) (80 nL L(-1)). Elevated O3 and CO2 caused extensive foliar injury of Dark Red Norland, but caused only slight injury of Superior. Elevated CO2 increased growth and tuber yield of both cultivars, whereas elevated O3 generally suppressed growth and yield, mainly of Dark Red Norland. Elevated CO2 appeared to protect Dark Red Norland from O3-induced suppression of shoot, root, and tuber weight as measured at midseason but did not protect either cultivar from O3 stress at the final harvest. The results further illustrate the difficulty in predicting effects of O3 + CO2 mixtures based on the effects of the individual gases. PMID- 14535301 TI - Atmospheric deposition of pesticides to an agricultural watershed of the Chesapeake Bay. AB - The Choptank River watershed, located on the Delmarva Peninsula of the Chesapeake Bay, is dominated by agricultural land use, which makes it vulnerable to runoff and atmospheric deposition of pesticides. Agricultural and wildlife areas are in close proximity and off-site losses of pesticides may contribute to toxic effects on sensitive species of plants and animals. High-volume air samples (n = 31) and event-based rain samples (n = 71) were collected from a single location in the watershed representing regional background conditions. Surface water samples were collected from eight stations in the tidal portion of the river on five occasions during 2000. Chlorothalonil, metolachlor, atrazine, simazine, endosulfan, and chlorpyrifos were frequently detected in the air and rain, with maximal concentrations during the period when local or regional crops were planted. The wet deposition load to the watershed was estimated at 150 +/- 16, 61 +/- 7, and 51 +/- 6 kg yr(-1) for chlorothalonil, metolachlor, and atrazine, respectively. The high wet deposition load compared with the estimated annual usage for chlorothalonil (13%) and endosulfan (14-90%) suggests an atmospheric source from outside the watershed. Net air-water gas exchange fluxes for metolachlor varied from -44 +/- 19 to 9.3 +/- 4.1 ng m(-2) d(-1) with negative values indicating net deposition. Wet deposition accounted for 3 to 20% of the total metolachlor mass in the Choptank River and was a more important source to the river than gas exchange. Estimates of herbicide flux presented here are probably a low estimate and actual rates may be significantly higher in areas closer to pesticide application. PMID- 14535302 TI - Predicting and measuring environmental concentration of pesticides in air after soil application. AB - Pesticides can volatilize into the atmosphere, which affects the air quality. The ability to predict pesticide volatilization is an essential tool for human risk and environmental assessment. Even though there are several mathematical models to assess and predict the fate of pesticides in different compartments of the environment, there is no reliable model to predict volatilization. The objectives of this study were to evaluate pesticide volatilization under agricultural conditions using malathion [ O,O-dimethyl-S-(1,2-dicarbethoxyethyl) dithiophosphate], ethoprophos (O-ethyl S,S-dipropylphosphorodithioate), and procymidone [N-(3,5-dichlorophenyl)-1,2-dimethylcyclopropane-1,2-dicarboximide] as test compounds and to evaluate the ability of the Pesticide Leaching Model (PELMO) to calculate the predicted environmental concentrations of pesticides in air under field conditions. The volatilization rate of procymidone, malathion, and ethoprophos was determined in a field study during two different periods (December 1998 and September 1999) using the Theoretical Profile Shape (TPS) method. The experiments were performed on bare silty soil in the Bologna province, Italy. Residues in the air were continuously monitored for 2 to 3 wk after the pesticide applications. The amount of pesticide volatilized was 16, 5, and 11% in December 1998 and 41, 23, and 19% in September 1999 for procymidone, malathion, and ethoprophos, respectively. In both these experiments, the PELMO simulations of the concentration of ethoprophos and procymidone were in good agreement with the measured data (factor +/- 1.1 on average). The volatilization of malathion was underestimated by a factor of 30 on average. These results suggest that volatilization described by PELMO may be reliable for volatile substances, but PELMO may underpredict volatilization for less-volatile substances. PMID- 14535303 TI - Simple protocols to determine dust potentials from cattle feedlot soil and surface samples. AB - Cattle feedlot dust is an annoyance and may be a route for nutrient transport, odor emission, and pathogen dispersion, but important environmental factors that contribute to dust emissions are poorly characterized. A general protocol was devised to test feedlot samples for their ability to produce dust under a variety of environmental conditions. A blender was modified to produce dust from a variety of dried feedlot surface and soil samples and collect airborne particles on glass fiber filters by vacuum collection. A general blending protocol optimized for sample volume (150-175 cm3), blending time (5 min of pre-blending), and dust collection time (15 s) provided consistent dust measurements for all samples tested. The procedure performed well on samples that varied in organic matter content, but was restricted to samples containing less than 200 to 700 g H2O kg(-1) dry matter (DM). When applied to field samples, the technique demonstrated considerable spatial variability between feedlot pen sites. Mechanistically, dust potential was related to moisture and organic matter content. An alternative protocol also demonstrated differences within pen sites in maximum dust potential and dust airborne residence time. The two protocols were not intended, nor are they suitable, for predicting actual particulate matter emissions from agricultural sources. Rather, the protocols rapidly and inexpensively compared the potential for dust emission from samples of differing composition under a variety of environmental conditions. PMID- 14535304 TI - In situ reduction of chromium(VI) in heavily contaminated soils through organic carbon amendment. AB - Chromium has become an important soil contaminant at many sites, and facilitating in situ reduction of toxic Cr(VI) to nontoxic Cr(III) is becoming an attractive remediation strategy. Acceleration of Cr(VI) reduction in soils by addition of organic carbon was tested in columns pretreated with solutions containing 1000 and 10 000 mg L(-1) Cr(VI) to evaluate potential in situ remediation of highly contaminated soils. Solutions containing 0,800, or 4000 mg L(-1) organic carbon in the form of tryptic soy broth or lactate were diffused into the Cr(VI) contaminated soils. Changes in Cr oxidation state were monitored through periodic micro-XANES analyses of soil columns. Effective first-order reduction rate constants ranged from 1.4 x 10(-8) to 1.5 x 10(-7) s(-1), with higher values obtained for lower levels of initial Cr(VI) and higher levels of organic carbon. Comparisons with sterile soils showed that microbially dependent processes were largely responsible for Cr(VI) reduction, except in the soils initially exposed to 10 000 mg L(-1) Cr(VI) solutions that receive little (800 mg L(-1)) or no organic carbon. However, the microbial populations (< or = 2.1 x 10(5) g(-1)) in the viable soils are probably too low for direct enzymatic Cr(VI) reduction to be important. Thus, synergistic effects sustained in whole soil systems may have accounted for most of the observed reduction. These results show that acceleration of in situ Cr(VI) reduction with addition of organic carbon is possible in even heavily contaminated soils and suggest that microbially dependent reduction pathways can be dominant. PMID- 14535305 TI - Removal of selenate in simulated agricultural drainage water by a rice straw bioreactor channel system. AB - Removal of selenium (Se) from agricultural drainage water is important in protecting wetland wildlife. Three flow-through bioreactor channel systems (BCSs), each with three channels filled with rice (Oryza sativa L.) straw, were set in the laboratory to determine removal of selenate [Se(VI)] (1020 microg L( 1)) from drainage water with a salinity of 10.4 dS m(-1), a pH of 8.1, and a nitrate (NO3-) range of 0 to 100 mg L(-1). Results showed that the rice straw effectively reduced Se(VI) during 122 to 165 d of the experiments. Calculation of Se mass in the three BCSs showed that 89.5 to 91.9% of the input Se(VI) was reduced to red elemental Se [Se(0)], where 96.6 to 98.2% was trapped in the BCSs. Losses of each gram of rice straw were almost equal to the removal of 1.66 mg of Se from the drainage water as a form of red Se(0), indicating that rice straw is a very effective organic source for removing Se(VI) from drainage water. PMID- 14535306 TI - Evaluation of water treatment sludge for ameliorating acid mine waste. AB - This study investigated the liming effect of water treatment sludge on acid mine spoils. The study was conducted with sludge from a water purification plant along the Vaal River catchments in South Africa. The optimum application rate for liming acid spoils and the speed and depth with which the sludge reacted with the mine waste were investigated. Chemical analysis indicated that the sludge is suitable as a liming agent because of its alkaline pH (8.08), high bicarbonate concentration (183.03 mg L(-1)), and low salinity (electrical conductivity = 76 mS m(-1)). The high cation exchange capacity of 15.47 cmol(c) kg(-1) and elevated nitrate concentration (73.16 mg L(-1)) also increase its value as an ameliorative material. The soluble concentrations for manganese, aluminum, lead, and selenium were high at a pH of 5 although only selenium (0.83 mg L(-1)) warranted some concern. According to experimental results, the application of 10 Mg ha(-1) of sludge to acid gold tailings increased the leach water pH from 4.5 to more than 7.5 and also increased the medium pH from 2.4 to 7.5. The addition of sludge further reduced the solubility of iron, manganese, copper, and zinc in the ameliorated gold tailings, but increased the electrical conductivity. The liming tempo was highest in the coal discard profile that had a coarse particle size distribution and took the longest to move through the gold tailings that had a fine particle size distribution. Results from this study indicate that the water treatment sludge investigated is suitable as a liming agent for rehabilitation of acid mine waste. PMID- 14535307 TI - The role of nitrilotriacetate in copper uptake by tobacco. AB - In growth chamber experiments we studied the effect of nitrilotriacetate (NTA) on Cu uptake by tobacco (Nicotiana tabacum L.). Plants were exposed for 6 d to 126 microM Cu and 500 microM NTA in nutrient solutions without and with 10 g L(-1) montmorillonite. Approximately seven times less Cu was dissolved in the montmorillonite solutions than in the nutrient solutions alone. In the absence of NTA, montmorillonite effectively competed with plant roots for Cu, although Cu remained bound to the roots. Nitrilotriacetate increased Cu uptake and translocation into shoots of tobacco by a factor of 3.5 from the nutrient solution and by a factor of 26 from the montmorillonite nutrient solution. Neither growth reduction nor any other visible sign of Cu toxicity was found in the presence of NTA with Cu concentrations of 190 mg kg(-1) in the shoots. In the absence of NTA, high Cu concentrations in root samples led to a brownish discoloration of the roots. PMID- 14535308 TI - Assessing the application of an additive model to estimate toxicity of a complex effluent. AB - A number of industries monitor levels of chemicals in their effluent, but few have undertaken prolonged biological monitoring of this wastewater. The focus of the present study was to determine whether past chemical data for effluent from a lead smelter could be used to estimate its past toxicity. Since the interactive effects of metals in effluents are often assumed to be additive, it was hypothesized that an additive model, 100/[sigma(metal concentration in effluent/EC50 for individual metal)], could be used to generate an EC50 from chemical data (where EC50 is the concentration of test material that affects 50% of the test organisms). To test the approach, a larval development toxicity test with the marine polychaete, Galeolaria caespitosa, was used to test 26 separate samples of effluent from a lead smelter, generating empirical EC50 values. EC50 values for each individual metal in the effluent were also generated using the larval development toxicity test. The concentrations of trace metals in each effluent sample were determined and, using the additive model, EC50 values were calculated. For the majority of effluent samples tested, the additive model underestimated toxicity, suggesting the presence of additional unidentified contaminants in the effluent samples. Additionally, a nonlinear rather than linear regression curve was found to best describe the relationship between the model and empirically derived EC50 values. This relationship was then used to estimate past trends in toxicity of the smelter effluent. Forty-eight percent of the variability in measured toxicity was explained by the model, with the model underestimating toxicity in the majority of samples. PMID- 14535309 TI - Sorption and desorption of cadmium by different fractions of biosolids-amended soils. AB - To evaluate the importance of both the inorganic and organic fractions in biosolids on Cd chemistry, a series of Cd sorption and desorption batch experiments (at pH 5.5) were conducted on different fractions of soils from a long-term field experimental site. The slope of the Cd sorption isotherm increased with rate of biosolids and was different for the different biosolids. Removal of organic carbon (OC) reduced the slope of the Cd sorption isotherm but did not account for the observed differences between biosolids-amended soils and a control soil, indicating that the increased adsorption associated with biosolids application was not limited to the increased OC from the addition of biosolids. Removal of both OC and Fe/Mn further reduced the slopes of Cd sorption isotherms and the sorption isotherm of the biosolids-amended soil was the same as that of the control, indicating both OC and Fe/Mn fractions added by the biosolids were important to the increased sorption observed for the biosolids amended soil samples. Desorption experiments failed to remove from 60 to 90% of the sorbed Cd. This "apparent hysteresis" was higher for biosolids-amended soil than the control soil. Removal of both OC and Fe/Mn fractions was more effective in removing the observed differences between the biosolids-amended soil and the control than either alone. Results show that Cd added to biosolids-amended soil behaves differently than Cd added to soils without biosolids and support the hypothesis that the addition of Fe and Mn in the biosolids increased the retention of Cd in biosolids-amended soils. PMID- 14535310 TI - Long-term effects of clipping and nitrogen management in turfgrass on soil organic carbon and nitrogen dynamics: the CENTURY model simulation. AB - Experiments to document the long-term effects of clipping management on N requirements, soil organic carbon (SOC), and soil organic nitrogen (SON) are difficult and costly and therefore few. The CENTURY ecosystem model offers an opportunity to study long-term effects of turfgrass clipping management on biomass production, N requirements, SOC and SON, and N leaching through computer simulation. In this study, the model was verified by comparing CENTURY-predicted Kentucky bluegrass (Poa pratensis L.) clipping yields with field-measured clipping yields. Long-term simulations were run for Kentucky bluegrass grown under home lawn conditions on a clay loam soil in Colorado. The model predicted that compared with clipping-removed management, returning clippings for 10 to 50 yr would increase soil C sequestration by 11 to 25% and nitrogen sequestration by 12 to 28% under a high (150 kg N ha(-1) yr(-1) nitrogen (N) fertilization regime, and increase soil carbon sequestration by 11 to 59% and N sequestration by 14 to 78% under a low (75 kg N ha(-1) yr(-1)) N fertilization regime. The CENTURY model was further used as a management supporting system to generate optimal N fertilization rates as a function of turfgrass age. Returning grass clippings to the turf-soil ecosystem can reduce N requirements by 25% from 1 to 10 yr after turf establishment, by 33% 11 to 25 yr after establishment, by 50% 25 to 50 yr after establishment, and by 60% thereafter. The CENTURY model shows potential for use as a decision-supporting tool for maintaining turf quality and minimizing negative environmental impacts. PMID- 14535311 TI - Kerogen in aquifer material and its strong sorption for nonionic organic pollutants. AB - Sorption of organic pollutants by subsurface materials has been found to not only correlate with the total organic carbon (TOC) content, but also depend on the types of soil and sediment organic matter (SOM). Characterization of geochemically heterogeneous SOM is key to elucidating sorption mechanisms and predicting pollutant transport in ground water systems. In this study, kerogen, a nonextractable organic material, was isolated with an acid demineralization procedure from a sandy aquifer material (Borden, Ontario, Canada) having a TOC content of approximately 0.021% (w/w). Petrographical examinations reveal that the kerogen has three major types of macerals including bituminite (Kerogen Type I and II), vitrinite (Type III), and fusinite (Type IV or charred kerogen). The solid-state 13C nuclear magnetic resonance (NMR) spectrum shows two dominant peaks, aliphatic and aromatic carbons, for the isolated material. Sorption isotherms measured using phenanthrene, naphthalene, 1,3,5-trichlorobenzene (TCB), and 1,2-dichlorobenzene (DCB) as sorbates showed that both the isolated kerogen and the original sand exhibited nonlinear sorption and that the phenanthrene and TCB isotherms measured for the kerogen material are more nonlinear than the respective isotherms for the original sand. The single-point organic carbon- normalized sorption capacity measured for the isolated kerogen can be several times greater than that measured for the original sand for a given sorbate. The study suggests that kerogen plays a major role in overall sorption isotherm nonlinearity and could yield higher-than-predicted sorption capacities for the subsurface material even though the content of this organic material is very low. PMID- 14535312 TI - Spatial distribution of DDT in sediments from estuarine rivers of central Florida. AB - Sediments may act as both a carrier for and a potential source of contaminants such as toxic organics in aquatic environments. This study investigated the spatial distribution of the pesticide DDT [1,1, 1-trichloro-2,2-bis(p chlorophenyl)ethane] in sediments from the Cedar and Ortega Rivers located in the lower St. Johns River basin, Florida, USA, using field measurements and three dimensional kriging analysis. High DDT concentrations were found near the junction of the Cedar and Ortega Rivers and at the north end of the Ortega River in the upper 0.5 m of the sediments, indicating that the sediment was enriched with DDT in the top layer although use of this chlorinated compound was banned in 1972. Further study revealed that the influence of sediment grain size or texture on DDT contamination was negligible in this river system and no linear correlations existed among DDT and its metabolites such as DDD [1,1-dichloro-2,2 bis(p-chlorophenyl)ethane] and DDE [1,1-dichloro-2,2-bis(p chlorophenyl)ethylene]. Comparison of three-dimensional distribution of DDT content to the Florida sediment quality assessment guideline or probable effect level (PEL) showed several "hot spots" in the Ortega River sediments, where DDT contents exceeded the PEL value of 4.78 microg kg(-1). Such contamination may pose a significant hazard to aquatic life. PMID- 14535313 TI - Pilot-scale treatment of RDX-contaminated soil with zerovalent iron. AB - Soils in Technical Area 16 at Los Alamos National Laboratory (LANL) are severely contaminated from past explosives testing and research. Our objective was to conduct laboratory and pilot-scale experiments to determine if zerovalent iron (Fe(0)) could effectively transform RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine) in two LANL soils that differed in physicochemical properties (Soils A and B). Laboratory tests indicated that Soil A was highly alkaline and needed to be acidified [with H2SO4, Al2(SO4)3, or CH3COOH] before Fe(0) could transform RDX. Pilot-scale experiments were performed by mixing Fe(0) and contaminated soil (70 kg), and acidifying amendments with a high-speed mixer that was a one-sixth replica of a field-scale unit. Soils were kept unsaturated (soil water content = 0.30-0.34 kg kg(-1)) and sampled with time (0-120 d). While adding CH3COOH improved the effectiveness of Fe(0) to remove RDX in Soil A (98% destruction), CH3COOH had a negative effect in Soil B. We believe that this difference is a result of high concentrations of organic matter and Ba. Adding CH3COOH to Soil B lowered pH and facilitated Ba release from BaSO4 or BaCO3, which decreased Fe(0) performance by promoting flocculation of humic material on the iron. Despite problems encountered with CH3COOH, pilot-scale treatment of Soil B (12 100 mg RDX kg(-1)) with Fe(0) or Fe(0) + Al2(SO4)3 showed high RDX destruction (96-98%). This indicates that RDX-contaminated soil can be remediated at the field scale with Fe(0) and soil-specific problems (i.e., alkalinity, high organic matter or Ba) can be overcome by adjustments to the Fe(0) treatment. PMID- 14535314 TI - Enhancing metolachlor destruction rates with aluminum and iron salts during zerovalent iron treatment. AB - Pesticide-contaminated soil may require remediation to mitigate ground and surface water contamination. We determined the effectiveness of zerovalent iron (Fe(0)) to dechlorinate metolachlor [2-chloro-N-(2-ethyl-6-methylphenyl)-N-(2 methoxy-1-methyl ethyl) acetamide] in the presence of aluminum and iron salts. By treating aqueous solutions of metolachlor with Fe(0), we found destruction kinetics were greatly enhanced when Al, Fe(II), or Fe(II) salts were added, with the following order of destruction kinetics observed: Al2(SO4)3 > AlCl3 > Fe2(SO4)3 > FeCl3. A common observation was the formation of green rusts, mixed Fe(II)-Fe(III) hydroxides with interlayer anions that impart a greenish-blue color. Central to the mechanism responsible for enhanced metolachlor loss may be the role these salts play in facilitating Fe(II) release. By tracking Al and Fe(II) in a Fe(0) + Al2(SO4)3 treatment of metolachlor, we observed that Al was readily sorbed by the corroding iron with a corresponding release of Fe(II). The manufacturing process used to produce the Fe(0) also profoundly affected destruction rates. Metolachlor destruction rates with salt-amended Fe(0) were greater with annealed iron (indirectly heated under a reducing atmosphere) than unannealed iron. Moreover, the optimum pH for metolachlor dechlorination in water and soil differed between iron sources (pH 3 for unannealed, pH 5 for annealed). Our results indicate that metolachlor destruction by Fe(0) treatment may be enhanced by adding Fe or Al salts and creating pH and redox conditions favoring the formation of green rusts. PMID- 14535315 TI - Competitive degradation between the fumigants chloropicrin and 1,3 dichloropropene in unamended and amended soils. AB - The mixture of 1,3-dichloropropene (1,3-D) and chloropicrin (CP) is used as a preplant soil fumigant. In comparison with individual fumigants, application of a mixture may affect the environmental dissipation and fate of each chemical, such as emission and degradation. We investigated the degradation of CP, 1,3-D, and their mixture in fresh soils and sterile soils, and evaluated the competitive characteristic of fumigants in the mixture. The degradation of low concentrations of CP in fresh soil was accelerated at early times in the presence of 1,3-D, whereas the addition of CP reduced the degradation rate of trans-1,3-D, possibly by inhibiting the activity of trans-1,3-D degrading microorganisms. The potential of applying amendments to the soil to increase the rate of CP and 1,3-D degradation was also illustrated. The degradation of both fumigants was significantly enhanced in soils amended with ammonium thiosulfate (ATS) and sodium diethyldithiocarbamate (Na-DEDTC) compared with unamended soil. Competitive degradation was observed for CP in amended soils in the presence of 1,3-D. The degradation of cis-1,3-D in amended soils spiked as a mixture of 1,3-D and CP was repressed compared with the rate of degradation in samples spiked with 1,3-D only. This implied that in abiotic degradation, CP and cis-1,3-D competed for a limited number of reaction sites in amended soil, resulting in decreased degradation rates. No significant influence of fumigant mixtures was observed for trans-1,3-D in amended soil. PMID- 14535316 TI - Organic manure and urea effect on metolachlor transport through packed soil columns. AB - Application of organic manure (OM) amendments and nitrogen fertilizers can affect the sorption and movement of pesticides in soil. This study summarizes the sorption and leaching of metolachlor [2-chloro-N-(2-ethyl-6-methylphenyl)-N-(2 methoxy-1-methylphenyl) acetamide] in soils after cow (Bos taurus) manure (2.5 and 5.0%) and urea (60 and 120 kg N ha(-1)) amendments in batch and column experiments. Both cow manure and urea applications increased metolachlor sorption in soils. The values of the Freundlich adsorption parameter K(r)(1/n) for treatments T0, T1 (OM), and T2 (OM) were 2.31, 3.32, and 3.96 in Soil 1; 2.02, 2.77, and 3.32 in Soil 2; and 1.10, 1.46, and 2.02 in Soil 3, respectively. Similarly, K(f)(1/n) values for treatment T1 (urea) and T2 (urea) were 2.37 and 2.84 in Soil 1; 2.16 and 2.83 in Soil 2; and 1.50 and 1.70 in Soil 3, respectively. Column leaching studies using Soil 1 indicated that OM application drastically reduced the metolachlor leaching losses from 50% (natural soil) to < 1.0% (5.0% OM amendment). Likewise, urea application also decreased metolachlor mobility and leaching losses in columns treated with 60 and 120 kg N ha(-1) urea were 33 and 20%, respectively. The reduction in the metolachlor leaching losses was achieved through the increase in the sorption capability of the OM- and urea amended soil. Therefore, coapplication of metolachlor with cow manure or urea fertilizers will not enhance metolachlor mobility and reduces metolachlor leaching losses in low-organic-matter soil. PMID- 14535317 TI - Phenanthrene sorption to structurally modified humic acids. AB - Several studies emphasize the importance of soil organic matter characteristics in hydrophobic contaminant sorption and outline the strong dependence of sorption on organic matter aromaticity. In this study, the role of organic matter aromaticity in phenanthrene sorption was investigated using humic acids (HAs) from compost, peat, and soil that were structurally modified by bleaching, hydrolysis, oximation, and subcritical water extraction. The HAs were characterized with cross polarization magic angle spinning carbon-13 nuclear magnetic resonance (CPMAS 13C NMR) spectroscopy and used in batch equilibrations with phenanthrene. Bleaching substantially reduced the aromaticity of the samples whereas the other treatments increased the relative aromaticity. Phenanthrene sorption increased, even though there was a substantial reduction in sorbent aromaticity with some samples. The HAs that exhibited comparable CPMAS 13C NMR spectra and aromaticity did not behave similarly with respect to phenanthrene sorption. When the sorption data (K(oc) values) were correlated to sample aromaticity, the correlation coefficients (r2) did not exceed 0.39. Comparisons with the atomic H to C ratio provided slightly better r2 values (up to 0.54). This study demonstrates that macroscopic sorbent characteristics could not explain the observed phenanthrene sorption coefficients, aliphatic structural components of HAs can contribute appreciably to phenanthrene sorption, and organic matter physical conformation may regulate access to organic matter structures. Therefore, the use of only macroscopic sorbent properties, such as aromaticity, to predict and rationalize sorption values cannot solely be used to explain the behavior of organic contaminants in soil environments. PMID- 14535318 TI - Uptake rates of thorium progeny in a semiarid environment. AB - The release rates and transformation processes that influence the mobility, biological uptake, and transfer of radionuclides are essential to the assessment of the health effects in the food chain and ecosystem. This study examined concentrations of 222Th in both soil and vegetation at a closed military training site, Kirtland Air Force Base (KAFB), New Mexico. Brazilian sludge was intentionally introduced into the topsoil in the early 1960s to simulate nuclear weapon accidents. Soil (60) and vegetation (120) samples were collected from 1996 to 2000 and analyzed for radionuclides and progeny. High-resolution gamma-ray spectroscopy was used to determine radionuclide activities. The results indicate that the thorium progeny were the predominant contaminant in soil and vegetation. Concentration ratios (CRs) were calculated based on actinium levels. PMID- 14535319 TI - Nitrate losses in subsurface drainage from a corn-soybean rotation as affected by time of nitrogen application and use of nitrapyrin. AB - Subsurface drainage, a water management practice used to remove excess water from poorly drained soils, can transport substantial amounts of NO3 from agricultural crop production systems to surface waters. A field study was conducted from the fall of 1986 through 1994 on a tile-drained Canisteo clay loam soil (fine-loamy, mixed, superactive, calcareous, mesic Typic Endoaquoll) to determine the influence of time of N application and use of nitrapyrin [NP; 2-chloro-6 (trichloromethyl) pyridine] on NO3 losses from a corn (Zea mays L.)-soybean [Glycine max (L.) Merr.] rotation. Four anhydrous ammonia treatments [fall N, fall N + NP, spring preplant N, and split N (40% preplant and 60% sidedress)] were replicated four times and applied at 150 kg N ha(-1) for corn on individual drainage plots. Sixty-two percent of the annual drainage and 69% of the annual NO3 loss occurred in April, May, and June. Flow-weighted NO3-N concentrations in the drainage water were two to three times greater in the two years following the three-year dry period compared with preceding and succeeding years. Nitrate N concentrations and losses in the drainage from corn were greatest for fall N with little difference among the other three N treatments. Nitrate losses from soybean were affected more by residual soil NO3 following corn than by the N treatments per se. Averaged across the four rotation cycles, flow-normalized NO3-N losses ranked in the order: fall N > split N > spring N = fall N + NP. Under these conditions NO3 losses from a corn-soybean rotation into subsurface drainage can be reduced by 13 to 18% by either applying N in the spring or using NP with late fall-applied ammonia. PMID- 14535320 TI - Biosolids decomposition after surface applications in west Texas. AB - In a semiarid environment, climate is a critical factor in the decomposition of surface-applied biosolids. This study examined the effect of 2- to 7-yr exposure times on the composition of single applications of New York, NY biosolids in western Texas. Exposure time effects on organic matter, N, P, S, Cu, Cr, Pb, Hg, and Zn were studied near Sierra Blanca, TX. Due to organic matter decomposition, total organic C decreased from 340 g kg(-1) in fresh biosolids to 180 g kg(-1) in biosolids after 82 mo of exposure, whereas the inorganic ash content of the biosolids increased from 339 to 600 g kg(-1). Total N decreased from 50 to 10 g N kg(-1) and total S decreased from 12 to 6 g S kg(-1). Bicarbonate-available P in the biosolids decreased from 0.9 to 0.2 g kg(-1). Successive H2O extractions yielded soluble P concentrations consistent with dicalcium phosphate (dical) for fresh biosolids and tricalcium phosphate (trical) for biosolids exposed for 59 months or more. Sparingly soluble phosphates, such as dical and trical, potentially yield > 0.5 mg P L(-1) in runoff waters for extended periods after biosolids applications, especially after multiple applications. Selective dissolution of the biosolids indicated that as much as 66 to 78% of P exists as iron phosphates, 16 to 21% as Fe oxides, and 5 to 12% as insoluble Ca phosphates. Chemical analyses of ash samples suggest that Cu and Zn have been lost from biosolids through leaching or runoff and no losses of Pb, Cr, or Hg have occurred since application. PMID- 14535322 TI - Stream transport of herbicides and metabolites in a tile-drained, agricultural watershed. AB - The occurrence of metabolites of many commonly used herbicides in streams has not been studied extensively in tile-drained watersheds. We collected water samples throughout the Upper Embarras River watershed [92% corn, Zea mays L., and soybean, Glycine max (L.) Merr.] in east-central Illinois from March 1999 through September 2000 to study the occurrence of atrazine (2-chloro-4-ethylamino-6 isopropylamino-s-triazine), metolachlor 12-chloro-N-(2-ethyl-6-methylphenyl)-N (methoxy-1-methylethyl) acetamide], alachlor [2-chloro-N-(2,6-diethylphenyl)-N (methoxymethyl) acetamide], acetochlor [2-chloro-N-(ethoxymethyl)-N-(2-ethyl-6 methylphenyl) acetamide], and their metabolites. River water samples were collected from three subwatersheds of varying tile density (2.8-5.3 km tile km( 2)) and from the outlet (United States Geological Survey [USGS] gage site). Near record-low totals for stream flow occurred during the study, and nearly all flow was from tiles. Concentrations of atrazine at the USGS gage site peaked at 15 and 17 microg L(-1) in 1999 and 2000, respectively, and metolachlor at 2.7 and 3.2 microg L(-1); this was during the first significant flow event following herbicide applications. Metabolites of the chloroacetanilide herbicides were detected more often than the parent compounds (evaluated during May to July each year, when tiles were flowing), with metolachlor ethanesulfonic acid [2-[(2-ethyl 6-methylphenyl)(2-methoxy-1-methylethyl)amino]-2-oxoethanesulfonic acid] detected most often (> 90% from all sites), and metolachlor oxanilic acid [2-[(2-ethyl-6 methylphenyl)(2-methoxy-1-methylethyl)amino]-2-oxoacetic acid] second (40-100% of samples at the four sites). When summed, the median concentration of the three chloroacetanilide parent compounds (acetochlor, alachlor, and metolachlor) at the USGS gage site was 3.4 microg L(-1), whereas it was 4.3 microg L(-1) for the six metabolites. These data confirm the importance of studying chloroacetanilide metabolites, along with parent compounds, in tile-drained watersheds. PMID- 14535321 TI - Manure history and long-term tillage effects on soil properties and phosphorus losses in runoff. AB - Manure additions to cropland can reduce total P losses in runoff on well-drained soils due to increased infiltration and reduced soil erosion. Surface residue management in subsequent years may influence the long-term risk of P losses as the manure-supplied organic matter decomposes. The effects of manure history and long-term (8-yr) tillage [chisel plow (CP) and no-till (NT)] on P levels in runoff in continuous corn (Zea mays L.) were investigated on well-drained silt loam soils of southern and southwestern Wisconsin. Soil P levels (0-15 cm) increased with the frequency of manure applications and P stratification was greater near the surface (0-5 cm) in NT than CP. In CP, soil test P level was linearly related to dissolved P (24-105 g ha(-1)) and bioavailable P (64-272 g ha(-1)) loads in runoff, but not total P (653-1893 g ha(-1)). In NT, P loads were reduced by an average of 57% for dissolved P, 70% for bioavailable P, and 91% for total P compared with CP. This reduction was due to lower sediment concentrations and/or lower runoff volumes in NT. There was no relationship between soil test P levels and runoff P concentrations or loads in NT. Long-term manure P applications in excess of P removal by corn in CP systems ultimately increased the potential for greater dissolved and bioavailable P losses in runoff by increasing soil P levels. Maintaining high surface residue cover such as those found in long-term NT corn production systems can mitigate this risk in addition to reducing sediment and particulate P losses. PMID- 14535323 TI - Irrigated mountain meadow fertilizer application timing effects on overland flow water quality. AB - Nonpoint-source pollution from agricultural activities is currently the leading cause of degradation of waterways in the United States. Applying best management practices to flood-irrigated mountain meadows may improve agricultural runoff and return flow water quality. Prior research has focused on fertilizer use for increased hay yields, while few studies have investigated the environmental implications of this practice. We examined the effects of fertilizer application timing on overland flow water quality from an irrigated mountain meadow near Gunnison, Colorado. Application of 40 kg phosphorus (P) and 19 kg nitrogen (N) ha(-1) using monoammonium phosphate (11-52-0, N-P-K) fertilizer to plots in the fall significantly reduced concentrations of reactive P and ammonium N in irrigation overland flow compared with early or late spring fertilization. Reactive P loading was 9 to almost 16 times greater when fertilizer was applied in the early or late spring, respectively, compared with in the fall. Ammonium N followed a similar trend with early spring loading more than 18 times greater and late spring loading more than 34 times greater than loads from fall-fertilized plots. Losses of 45% of the applied P and more than 17% of the N were measured in runoff when fertilizer was applied in the late spring. These results, coupled with those from previous studies, suggest that mountain meadow hay producers should apply fertilizer in the fall, especially P-based fertilizers, to improve hay yields, avoid economic losses from loss of applied fertilizers, and reduce the potential for impacts to water quality. PMID- 14535325 TI - Spatial and temporal patterns of Campylobacter contamination underlying public health risk in the Taieri River, New Zealand. AB - New Zealand's freshwater ecosystems are subject to microbial contamination from a predominantly agricultural landscape. This study examines the spatial and temporal distribution of the human pathogen Campylobacter in the lower Taieri River, South Island (New Zealand). Enumeration of thermophilic Campylobacter from river samples was performed using a most probable number (MPN) method. Seasonal variation in Campylobacter levels was evident, with higher median levels detected in summer, when human exposure through recreational water use is maximal. Campylobacter levels varied significantly among the 10 sampling sites, increasing below a major tributary entering the river and then showing a downstream decrease. These changes probably resulted from inputs from adjacent farms and instream Campylobacter losses (settling, death). Two main peaks in the flux of Campylobacter were observed, one in winter and one in summer. A decrease in notified cases of campylobacteriosis in the human population was observed when levels of Campylobacter at the main recreational bathing site on the river were low. Continuing land use change and intensification in New Zealand may lead to further increases in microbial contamination of freshwaters, and an associated increase in waterborne enteric diseases such as campylobacteriosis. PMID- 14535324 TI - Speciation of phosphorus in phosphorus-enriched agricultural soils using X-ray absorption near-edge structure spectroscopy and chemical fractionation. AB - Knowledge of phosphorus (P) species in P-rich soils is useful for assessing P mobility and potential transfer to ground water and surface waters. Soil P was studied using synchrotron X-ray absorption near-edge structure (XANES) spectroscopy (a nondestructive chemical-speciation technique) and sequential chemical fractionation. The objective was to determine the chemical speciation of P in long-term-fertilized, P-rich soils differing in pH, clay, and organic matter contents. Samples of three slightly acidic (pH 5.5-6.2) and two slightly alkaline (pH 7.4-7.6) soils were collected from A or B horizons in two distinct agrosystems in the province of Quebec, Canada. The soils contained between 800 and 2100 mg total P kg(-1). Distinct XANES features for Ca-phosphate mineral standards and for standards of adsorbed phosphate made it possible to differentiate these forms of P in the soil samples. The XANES results indicated that phosphate adsorbed on Fe- or Al-oxide minerals was present in all soils, with a higher proportion in acidic than in slightly alkaline samples. Calcium phosphate also occurred in all soils, regardless of pH. In agreement with chemical fractionation results, XANES data showed that Ca-phosphates were the dominant P forms in one acidic (pH 5.5) and in the two slightly alkaline (pH 7.4 7.6) soil samples. X-ray absorption near-edge structure spectroscopy directly identified certain forms of soil P, while chemical fractionation provided indirect supporting data and gave insights on additional forms of P such as organic pools that were not accounted for by the XANES analyses. PMID- 14535326 TI - Impact of dissolved organic matter on copper mobility in aquifer material. AB - Naturally occurring dissolved organic matter (DOM) and biosolids-derived DOM have been implicated in the mobility of metals in soils and aquifer materials. To investigate the effect of DOM on copper mobility in aquifer material, DOM derived from sewage biosolids was separated into two apparent molecular-weight (MW) fractions, 500 to 3500 Da (LMW) and > 14 000 Da (HMW). In each MW fraction, the DOM was further fractionated into hydrophilic, hydrophobic acid, and hydrophobic neutral compounds by an XAD-8 chromatography technique. The mobility of these DOM components and their influences on copper transport in a sesquioxide-coated, sandy aquifer material were examined with column transport experiments. The LMW DOM was found to be highly mobile, whereas the HMW DOM had a greater tendency to be retained by the aquifer material. Within the same MW fraction, the mobility of DOM followed the order of hydrophilic DOM > hydrophobic acid DOM > hydrophobic neutral DOM. Copper breakthrough curves in the presence of various DOM components showed that, except for the HMW hydrophilic fraction, DOM components enhanced Cu transport through the aquifer columns at early stages of transport (the first 75 pore volumes). In the later stages, however, all the DOM components substantially inhibited Cu mobility. We hypothesize that several mechanisms could account for retardation of Cu movement in the presence of the DOM fractions, including the formation of ternary complexes between the aquifer material, Cu, and DOM; changes in the electrostatic potential at the solid-phase surface; and pH buffering by DOM. PMID- 14535327 TI - Electromagnetic induction methods applied to an abandoned manure handling site to determine nutrient buildup. AB - Movement of nutrients from livestock manure handling sites has the potential to negatively impact the environment. This study was conducted using electromagnetic induction (EMI) measurements to develop apparent soil electrical conductivity (EC(a)) maps to identify regions of nutrient buildup beneath an abandoned compost site. A trailer-mounted EM-38 coupled with a global positioning satellite system was towed across an area used for composting of feedlot manure. The resulting EC(a) maps were compared with known locations of compost rows confirming the alignment of row locations with high EC(a) regions. The identified rows were cored and compared with the region between the rows. The identified rows with a compost history demonstrated significant (P < 0.05) increases in soluble salts (1.6 times greater), NO3 (6.0 times greater), and Cl (2.0 times greater) compared with the area between the rows at a 1.5-m depth. Image processing techniques were used to display yearly changes that were associated with nutrient movement and transformations in the soil beneath the site. Correlations between EMI measurements and soil core analyses for NO3-N, Cl, and EC provided ancillary support for the EMI methods. The use of EMI for mapping of sites having a history of livestock waste application was effective in delineating high nutrient buildup areas and for observing spatial EC(a) changes over time. PMID- 14535328 TI - Nitrogen management considerations for landspreading municipal solid waste compost. AB - Many municipalities have examined composting as an alternative to landfilling for the management of organic solid waste materials. Ultimately these materials will be land-applied and therefore some knowledge of nutrient availability will be necessary to optimize crop yield and minimize environmental risk. Field studies were conducted in 1993 and 1994 on a silt loam and a loamy sand soil in Wisconsin to determine the effect of municipal solid waste compost (MSWC) on corn (Zea mays L.) yield, plant nutrient concentration, and soil nitrate N content. Municipal solid waste composts with ages of 7, 36, and 270 d were applied at rates of 22.5, 45, and 90 Mg ha(-1) to small plots. Rates of commercial nitrogen (N) fertilizer, ranging from 0 to 179 kg N ha(-1), were applied to separate plots to determine the N availability from the MSWC. Treatments were applied in the spring and incorporated before planting corn. The 270-d MSWC increased corn whole-plant dry matter and grain yield at each location in both years above the 7- and 36-d MSWC. Rate of MSWC only affected grain yield at the loamy sand site in 1994. Municipal solid waste compost had minimal effect on the levels of plant nutrients in the whole-plant tissue measured at physiological maturity. Nitrate N measured in the top 90 cm of soil was higher throughout the growing season in treatments receiving recommended N fertilizer when compared with any of the MSWC treatments. It was estimated that 6 to 17% of the total N in the 270-d MSWC became available in the first year. The land-application of mature MSWC at the tested rates would be an agronomically and environmentally admissible practice. PMID- 14535329 TI - Biosolids-derived nitrogen mineralization and transformation in forest soils. AB - Utilization of biosolids through land application is becoming increasingly popular among wastewater managers. To minimize the potential contamination of receiving waters from biosolids-derived nitrogen (N), it is important to understand the availability of N after land application of biosolids. In this study, four secondary biosolids (two municipal and two pulp and paper industrial biosolids) were used in a laboratory incubation experiment to simulate N mineralization and transformation after land application. Municipal biosolids were from either aerobically or anaerobically digested sources, while pulp and paper industrial biosolids were from aerated wastewater stabilization lagoons. These biosolids were mixed with two New Zealand forest soils (top 100 mm of a volcanic soil and a brown soil) and incubated at two temperatures (10 and 20 degrees C) for 26 wk. During incubation, mineralized N was periodically leached from the soil-biosolids mixture with 0.01 M CaCl2 solution and concentrations of NH4 and NO3 in leachate were determined. Mineralization of N from aerobically digested municipal biosolids (32.1%) was significantly more than that from anaerobically digested biosolids (15.2%). Among the two pulp and paper industrial biosolids, little N leached from one, while as much as 18.0% of total organic N was leached from the other. As expected, mineralization of N was significantly greater at 20 degrees C (average 22.8%) than at 10 degrees C (average 9.7%). It was observed that more N in municipal biosolids was mineralized in the brown soil, whereas more N in pulp and paper industrial biosolids mineralized in the volcanic soil. Transformation of NH4 to NO3 was affected by soil type and temperature. PMID- 14535330 TI - Phosphorus retention mechanisms of a water treatment residual. AB - Water treatment residuals (WTRs) are a by-product of municipal drinking water treatment plants and can have the capacity to adsorb tremendous amounts of P. Understanding the WTR phosphorus adsorption process is important for discerning the mechanism and tenacity of P retention. We studied P adsorbing mechanism(s) of an aluminum-based [Al2(SO4)3 x 14H2O] WTR from Englewood, CO. In a laboratory study, we shook mixtures of P-loaded WTR for 1 to 211 d followed by solution pH analysis, and solution Ca, Al, and P analysis via inductively coupled plasma atomic emission spectroscopy. After shaking periods, we also examined the solids fraction by X-ray diffraction (XRD) and electron microprobe analysis using wavelength dispersive spectroscopy (EMPA-WDS). The shaking results indicated an increase in pH from 7.2 to 8.2, an increase in desorbed Ca and Al concentrations, and a decrease in desorbed P concentration. The pH and desorbed Ca concentration increases suggested that CaCO3 controlled Ca solubility. Increased desorbed Al concentration may have been due to Al(OH)4 formation. Decreased P content, in conjunction with the pH increase, was consistent with calcium phosphate formation or precipitation. The system appeared to be undersaturated with respect to dicalcium phosphate (DCP; CaHPO4) and supersaturated with respect to octacalcium phosphate [OCP; Ca4H(PO4)3 x 2.5H2O]. The Ca and Al increases, as well as OCP formation, were supported by MINTEQA2 modeling. The XRD and EMPA-WDS results for all shaking times, however, suggested surface P chemisorption as an amorphous Al P mineral phase. PMID- 14535331 TI - Rainfall timing and frequency influence on leaching of Escherichia coli RS2G through soil following manure application. AB - The time between swine (Sus scrofa) manure application to soil as a crop fertilizer, the first rainfall event, and the frequency of rainfall events should influence leaching potential of fecal pathogens. Soil microcosms were inoculated in the lab with a swine manure isolate of Escherichia coli, strain RS2G, expressing green fluorescent protein, to examine how timing and frequency of rainfall events influences RS2G leaching and survival in soil. Liquid swine manure inoculated with RS2G was applied to intact soil cores (20 cm in diameter x 30 cm long) 4, 8, or 16 d before the first rainfall event (50.8 mm over a 4-h period), and each core received one to three rainfall events. Manure application methods (no-till surface-broadcast, broadcast and incorporated, and tilled before broadcast) had no affect on leaching, although there was greater survival in soils when the manure had been incorporated. Most of the RS2G in the leachate appeared following the first rainfall event and RS2G leaching decreased with increasing time between manure application and the first rainfall, although leachates contained as much as 3.4 to 4.5 log colony forming units (CFU) 100 mL( 1) of RS2G when the first rainfall occurred 16 d after manure application. With increasing frequency of rainfalls there was a decrease in subsequent concentrations of RS2G in the leachate. There was no correlation between leachate RS2G and total coliforms or fecal streptococci concentrations. Soil RS2G numbers were 1 to 10% of the inoculum regardless of the length of time between manure application and the first rainfall. RS2G leaching was mostly influenced by the time between manure application and first rainfall event, and significant leaching and survival in soil was possible even if the first rain occurred 16 d after manure application. PMID- 14535332 TI - On-farm assessment of biosolids effects on soil and crop tissue quality. AB - Agronomic use of biosolids as a fertilizer material remains controversial in part due to public concerns regarding the potential pollution of soils, crop tissue, and ground water by excess nutrients and trace elements in biosolids. This study was designed to assess the effects of long-term commercial-scale application of biosolids on soils and crop tissue sampled from 18 production farms throughout Pennsylvania. Biosolids application rates ranged from 5 to 159 Mg ha(-1) on a dry weight basis. Soil cores and crop tissue samples from corn (Zea mays L.), soybean (Glycine spp.), alfalfa (Medicago sativa L.), orchardgrass (Dactylis spp.) hay, and/or sorghum [Sorghum bicolor (L.) Moench] were collected for three years from georeferenced locations at each farm. Samples were tested for nutrients, trace elements, and other variables. Biosolids-treated fields had more post-growing season soil NO3 and Ca and less soil K than control fields and there was some evidence that soil P concentrations were higher in treated fields. The soil concentrations of Cu, Cr, Hg, Mo, Mn, Pb, and Zn were higher in biosolids-treated fields than in control fields; however, differences were < or = 0.06 of the USEPA Part 503 cumulative pollutant loading rates (CPLRs). There were no differences in the concentrations of measured nutrients or trace elements in the crop tissue grown on treated or control fields at any time during the study. Commercial-scale biosolids application resulted in soil trace element increases that were in line with expected increases based on estimated trace element loading. Excess NO3 and apparent P buildup indicates a need to reassess biosolids nutrient management practices. PMID- 14535333 TI - Recycling soil nitrate nitrogen by amending agricultural lands with oily food waste. AB - With current agricultural practices the amounts of fertilizer N applied are frequently more than the amounts removed by the crop. Excessive N application may result in short-term accumulation of nitrate nitrogen (NO3-N) in soil, which can easily be leached from the root zone and into the ground water. A management practice suggested for conserving accumulated NO3-N is the application of oily food waste (FOG; fat + oil + greases) to agricultural soils. A two-year field study (1995-1996 and 1996-1997) was conducted at Elora Research Center (43 degrees 38' N, 80 degrees W; 346 m above mean sea level), University of Guelph, Ontario, Canada to determine the effect of FOG application in fall and spring on soil NO3-N contents and apparent N immobilization-mineralization of soil N in the 0- to 60-cm soil layer. The experiment was planned under a randomized complete block design with four replications. An unamended control and a reference treatment [winter wheat (Triticum aestivum L.) cover crop] were included in the experiment to compare the effects of fall and spring treatment of oily food waste on soil NO3-N contents and apparent N immobilization-mineralization. Oily food waste application at 10 Mg ha(-1) in the fall decreased soil NO3-N by immobilization and conserved 47 to 56 kg NO3-N ha(-1), which would otherwise be subject to leaching. Nitrogen immobilized due to FOG application in the fall was subsequently remineralized by the time of fertilizer N sidedress, whereas no net mineralization was observed in spring-amended plots at the same time. PMID- 14535335 TI - Particulates, not plants, dominate nitrogen processing in a septage-treating aerated pond system. AB - In pond and wetland systems for wastewater treatment, plants are often thought to enhance the removal of ammonium and nitrogen through the activities of root associated bacteria. In this study, we examined the role of plant roots in an aerated pond system with floating plants designed to treat high-strength septage wastewater. We performed both laboratory and full-scale experiments to test the effect of different plant root to septage ratios on nitrification and denitrification, and measured the abundances of nitrifying bacteria associated with roots and septage particulates. Root-associated nitrifying bacteria did not play a significant role in ammonium and total nitrogen removal. Investigations of nitrifier populations showed that only 10% were associated with water hyacinth [Eichhornia crassipes (Mart.) Solms] roots (at standard facility plant densities equivalent to 2.2 wet g roots L(-1) septage); instead, nitrifiers were found almost entirely (90%) associated with suspended septage particulates. The role of root-associated nitrifiers in nitrification was examined in laboratory batch experiments where high plant root concentrations (7.4 wet g L(-1), representing a 38% net increase in total nitrifier populations over plant-free controls) yielded a corresponding increase (55%) in the non-substrate-limited nitrification rate (V(max)). However, within the full-scale septage-treating pond system, nitrification and denitrification rates remained unchanged when plant root concentrations were increased to 7.1 g roots L(-1) (achieved by increasing the surface area available for plants while maintaining the same tank volume). Under normal facility operating conditions, nitrification was limited by ammonium concentration, not nitrifier availability. Maximizing plant root concentrations was found to be an inefficient mechanism for increasing nitrification in organic particulate-rich wastewaters such as septage. PMID- 14535334 TI - Bedding and seasonal effects on chemical and bacterial properties of feedlot cattle manure. AB - Nutrients, soluble salts, and pathogenic bacteria in feedlot-pen manure have the potential to cause pollution of the environment. A three-year study (1998-2000) was conducted at a beef cattle (Bos taurus) feedlot in southern Alberta, Canada to determine the effect of bedding material [barley (Hordeum vulgare L.) straw versus wood chips] and season on the chemical and bacterial properties of pen floor manure. Manure was sampled for chemical content (N, P, soluble salts, electrical conductivity, and pH) and populations of four groups of bacteria (Escherichia coli, total coliforms, and total aerobic heterotrophs at 27 and 39 degrees C). More chemical parameters of manure were significantly (P < or = 0.05) affected by season (SO4, Na, Mg, K, Ca, sodium adsorption ratio [SAR], total C, NO3-N, NH4-N, total P, and available P) than by bedding (K, pH, total C, C to N ratio, NH4-N, and available P). Bedding had no significant (P > 0.05) effect on the four bacterial groups whereas season affected all four groups. Numbers of E. coli and total coliforms (TC) were significantly higher by 1.72 to 2.02 log10 units in the summer than the other three seasons, which was consistent with a strong positive correlation of E. coli and TC with air temperature. The low ratio of bedding to manure in the pens was probably the major cause of the lack of significant bedding effects. Bedding material and seasonal timing of cleaning feedlot pens and land application of manure may be a potential tool to manage nutrients, soluble salts, and pathogens in manure. PMID- 14535336 TI - Corer-reactors for contaminant flux measurement in sediments. AB - Design details and operating instructions are provided for a sediment corer that can be converted into a reactor for the measurement of the fluxes of contaminants from sediments to overlying waters. The corer-reactor permits measurements, under controlled laboratory conditions, on intact, largely undisturbed sediment cores, without significantly perturbing the physical and chemical conditions found in the field. The design can be constructed in-house for around US dollar 240 (A dollar 400) (excluding motor and corer lid), making it a relatively inexpensive system. PMID- 14535337 TI - Cadmium availability from granulated and bulk-blended phosphate-potassium fertilizers. AB - Recent field experiments have shown that high chloride (Cl) in irrigation waters can increase soil cadmium (Cd) uptake by crops because of the formation of soluble ion-pair complexes of Cd with Cl in soil solution. The present study was aimed at testing a hypothesis that KCl in granulated PK fertilizers may enhance Cd uptake by crops from Cd-containing P fertilizers because of close contact between Cd and Cl in the same granules. Less effect would be expected if the same granular PK fertilizers were bulk-blended because of separation of Cd and Cl in different granules. A single superphosphate (SSP) containing 32 mg Cd kg(-1) was granulated by the compaction process with KCl at a P to K ratio = 1:1. Granular KCl was also bulk-blended with granular SSP or Cd-free monocalcium phosphate (MCP) at the same P to K ratio. An acid Ultisol (pH 5.2) was treated with PK fertilizers at 400 mg kg(-1) each for P and K. Upland rice (Oryza sativa L.) and soybean [Glycine max (L.) Merr.] were grown to maturity, and signalgrass (Brachiaria decumbens Stapf.) was cut four times during the study. The results showed that the agronomic effectiveness in increasing crop yield was the same with SSP and MCP whether granulated or bulk-blended with KCl. Concentrations of Cd in plant tissue samples of all crops were much lower for MCP than for SSP. In all the plant tissue samples, except grain samples of upland rice, Cd concentrations obtained with granulated (SSP + KCl) were significantly higher than that with bulk-blended (SSP) + (KCl). PMID- 14535338 TI - Gas-phase sorption-desorption of propargyl bromide and 1,3-dichloropropene on plastic materials. AB - The goal of this research was to provide information for choosing appropriate materials for studying gas-phase concentrations of propargyl bromide (3BP) and 1,3-dichloropropene (1,3-D) in laboratory experiments. Several materials were tested and found to sorb both gas-phase chemicals in the following order: stainless steel (SS) < Teflon polytetrafluoroethylene (PTFE-FEP) approximately flexible polyvinyl chloride (PVC) approximately acrylic < low-density polyethylene (PE) < vinyl approximately silicone < polyurethane foam (PUF). Sorption of SS was insignificant and PUF sorbed all the fumigant that was applied. For the other materials, linear sorption coefficients (Kd) for 3BP ranged from 3.0 cm3 g(-1) for PVC to 215 cm3 g(-1) for silicone. Freundlich sorption coefficients for 1,3-D ranged from 11.5 to 371 cm3 g(-1). First-order desorption rate constants in an open system ranged from 0.05 to 1.38 h(-1) for 3BP and from 0.07 to 1.73 h(-1) for 1,3-D. In a closed system, less than 2% of sorbed fumigant desorbed from vinyl while up to 99% desorbed from PVC within 24 h when equilibrated at the highest headspace concentration. Sorption of both fumigants was linearly related to the square root of time except for vinyl and silicone. This may indicate non-fickian diffusion of fumigant into the polymer matrix. Vinyl, silicone, PE, and PUF should be avoided for quantitative study of organic gases, except possibly as a trapping medium. Use of PTFE, PVC, and acrylic may require correction for sorption-desorption and diffusion. PMID- 14535339 TI - Determination of polyacrylamide in soil waters by size exclusion chromatography. AB - Determination of polyacrylamide (PAM) concentration in soil waters is important in improving the efficiency of PAM application and understanding the environmental fate of applied PAM. In this study, concentrations of anionic PAM with high molecular weight in soil waters containing salts and dissolved organic matter (DOM) were determined quantitatively by size exclusion chromatography (SEC) with ultraviolet (UV) absorbance detection. Polyacrylamide was separated from interferential salts and DOM on a polymeric gel column eluted with an aqueous solution of 0.05 M KH2PO4 and then detected at a short UV wavelength of 195 nm. Analysis of PAM concentrations in soil sorption supernatants, soil leachates, and water samples from irrigation furrow streams showed that SEC is an effective approach for quantifying low concentrations (0-10 mg L(-1)) of PAM in waters containing soil DOM and salts. The method has a lower detection limit of 0.02 microg and a linear response range of 0.2 to 80 mg L(-1). Precision studies gave coefficients of variation of < 1.96% (n = 4) for > 10 mg L(-1) PAM and < 12% (n = 3) for 0.2 to 3 mg L(-1) PAM. PMID- 14535340 TI - Nuclearia pattersoni sp. n. (Filosea), a new species of amphizoic amoeba isolated from gills of roach (Rutilus rutilus), and its rickettsial endosymbiont. AB - A new species of amphizoic amoeba, Nuclearia pattersoni sp. n., isolated from gills of Rutilus rutilus L. is described. It is characterised by elongate flattened trophozoites of irregular shape. The longer dimension of their bodies is 13.2 (11.0-15.7) microm. Filopodia radiating mostly from the poles are 2 to 2.5 times longer than the body. The diameter of less frequently observed spherical trophozoites is 8.2-10.8 microm; their filopodia radiate to all directions. Cyst-like stages have shorter pseudopodia that arise from one pole only. The surface of locomotive forms from agar plate cultures has a thin amorphous glycocalyx, while most cells are covered by two layers of extracellular matrix. Mitochondria have flattened cristae, dictyosomes are located in the perinuclear zone. A conspicuous ultrastructural feature of the morphologically similar N. simplex, perinuclear striated band, is not present. Light microscopic and ultrastructural data are completed with the sequence of SSU rRNA gene and phylogenetic analysis including sequences of related taxa. The bacterial endosymbiont found in N. pattersoni type strain RR2G2 is assigned to the genus Rickettsia. PMID- 14535341 TI - Life cycle and epizootiology of Amblyospora ferocis (Microspora: Amblyosporidae) in the mosquito Psorophora ferox (Diptera: Culicidae). AB - A natural population of Psorophora ferox (Humbold, 1820) infected with the microsporidium Amblyospora ferocis Garcia et Becnel, 1994 was sampled weekly during a seven-month survey in Punta Lara, Buenos Aires Province, Argentina. The sequence of development of A. ferocis in larvae of P. ferox leading to the formation of meiospores followed the developmental pathway previously reported for various species of Amblyospora. The natural prevalence of A. ferocis in the larval population of P. ferox ranged from 0.4% to 13.8%. Spores were detected in the ovaries of field-collected females of P. ferox and were shown to be responsible for transovarial transmission of A. ferocis to the next generation of mosquito larvae in laboratory tests. These spores were binucleate and slightly pyriform in shape. The prevalence of A. ferocis in the adult population ranged from 2.7% to 13.9%. Data on effects of the infection on female fecundity showed that infected field-collected adults of P. ferox laid an average of 47.6 +/- 6.5 eggs of which 35.8% +/- 4.1% hatched. Uninfected field-collected adults of P. ferox laid 82.8 +/- 6.8 eggs of which 64.1% +/- 5.5% hatched. Six species of copepods living together with P. ferox were fed meiospores from field-infected larvae but none became infected. Horizontal transmission of A. ferocis to P. ferox larvae remains unknown. PMID- 14535342 TI - Rheumatoid factor-like IgM in Plasmodium berghei (Apicomplexa: Haemosporida) infections of BALB/c mice. AB - Groups of female BALB/c mice infected by intravenous injection with 50 erythrocytes containing Plasmodium berghei Vincke et Lips, 1948 were sacrificed on days 3 through 12 after infection. Rheumatoid factor-like IgM (RF-IgM) and parasite-specific IgG levels were determined by enzyme-linked immunosorbent assay in serum specimens and in culture medium removed from spleen cell cultures established at sacrifice. All four mouse IgG subisotypes were recognized by RF IgM molecules induced by Plasmodium berghei infection, and in this regard, the parasite-induced RF-IgM response resembled that induced by lipopolysaccharide polyclonal activation. Plasmodium berghei infection resulted in a biphasic RF-IgM response, with infected animals demonstrating significantly increased levels of RF-IgM early in the infection and significantly decreased levels late in the infection, compared to uninfected control mice. The decreased levels of RF-IgM observed late in infection correlated with increasing parasitaemia levels, and were primarily due to a decrease in RF-IgM specific for mouse IgG2a. Late infection levels of RF-IgM specific for IgGI, IgG2b, and IgG3 were not significantly different from those of control animals. PMID- 14535343 TI - Myxozoans infecting the sharptooth catfish, Clarias gariepinus in the Okavango River and Delta, Botswana, including descriptions of two new species, Henneguya samochimensis sp. n. and Myxobolus gariepinus sp. n. AB - During a recent investigation of parasites infecting fishes from the Okavango River and Delta, Botswana (southern Africa) fourteen sharptooth catfish, Clarias gariepinus (Burchell, 1822) (Siluriformes: Clariidae) were examined for the presence of myxozoan infections. Results revealed the presence of two species of the genus Henneguya Thelohan, 1895 and one species of the genus Myxobolus Butschli, 1882 infecting this fish host. Two of the sampled fish exhibited large plasmodia of Henneguya suprabranchiae Landsberg, 1987 in the cartilage of the accessory breathing organ, another two individuals were infected with H. samochimensis sp. n. plasmodia in the gills and another three individuals revealed an infection with Myxobolus gariepinus sp. n. plasmodia in the ovaries. PMID- 14535344 TI - Myxidium biliare sp. n. (Myxozoa) from gall bladder of Galaxias maculatus (Osmeriformes: Galaxiidae) in Patagonia (Argentina). AB - Myxidium biliare sp. n., a new myxosporean species parasitizing the gall bladder of Galaxias maculatus (Jenyns), in Patagonia, is described. Its coelozoic plasmodia were floating free in the bile. Spores are fusiform 13.7 +/- 0.9 microm long and 6.9 +/- 0.6 microm wide, with rounded ends in frontal view and slightly pointed ends in sutural view; shell with ridges and sinuous sutural line. Both maximum prevalence and maximum percentage of immature plasmodia occurred in summer. In winter the prevalence and the percentage of immature plasmodia fell to their lowest values. Prevalence was independent of host sex but increased with host length. Prevalence in 15 Patagonian Andean lakes (situated from 39 degrees 25'S to 41 degrees 30'S) ranged between 4.2% and 70%. PMID- 14535345 TI - Four new species of Cichlidogyrus (Monogenea: Ancyrocephalidae), gill parasites of Tilapia cabrae (Teleostei: Cichlidae), with discussion on relative length of haptoral sclerites. AB - Tilapia cabrae Boulenger, 1899, a cichlid fish from coastal lowlands of the Republic of Congo (Africa), was examined for gill parasites. Four new species of the Monogenea were found, all belonging to Cichlidogyrus (Ancyrocephalidae): C. berradae sp. n., C. revesati sp. n., C. legendrei sp. n., and C. lemoallei sp. n. A possible relationship between the relative size of haptoral sclerites (i.e., uncinuli compared to gripi) and microhabitat selection in the studied host parasite model is discussed. PMID- 14535346 TI - Three species of Magnibursatus Naidenova, 1969 (Digenea: Derogenidae) from Atlantic and Black Sea marine teleosts. AB - Three species of Magnibursatus Naidenova, 1969 are described from marine teleosts: M. skrjabini (Vlasenko, 1931), the type species of the genus, from the gobiid Zosterisessor ophiocephalus on the Bulgarian Black Sea coast; M. bartolii sp. n. from the sparid Boops boops off the Atlantic coast of Spain; and M. minutus sp. n. from the gobiid Neogobius eurycephalus on the Bulgarian Black Sea coast. M. bartolii differs from all other Magnibursatus species in its larger sinus-sac (length > 250 microm, width > 150 microm) and the more posterior location of testes. This species is also unusual in that it occurs in the branchial chamber and on the gills of its host. M. minutus is distinguished by the distinctly smaller dimensions of the body (length < 1000 microm, width < 200 microm), organs and eggs. These species are also distinguished from both M. caudofilamentosa (Reimer, 1971) and Tyrrhenia blennii Paggi et Orecchia, 1975. A key to the species of Magnibursatus is presented. PMID- 14535347 TI - Posterotestes unelen gen. et sp. n. (Digenea: Apocreadiidae) from Percichthys trucha (Osteichthyes: Percichthyidae) in Patagonia, Argentina. AB - Posterotestes gen. n. (Digenea: Apocreadiidae, Apocrcadiinae) is proposed for specimens with the following features: spiny body, posterior position of gonads, extension of vitelline follicles up to level of intestinal bifurcation and absent at post-testicular space, long oesophagus and extension of caeca up to anterior testis. Posterotestes unelen sp. n. is described from the intestine of the native fish, Percichthys trucha (Cuvier et Valenciennes) (Osteichthyes: Percichthyidae) from Patagonian Andean lakes. PMID- 14535348 TI - Transmission electron microscopy of intra-tegumental sensory receptors in the forebody of Crepidostomum metoecus (Digenea: Allocreadiidae). AB - In the adult fish trematode Crepidostomum metoecus (Braun, 1900), four types of sensory receptors were observed inside the forebody tegument and one type beneath the tegument basal lamina. Two types of sensory receptors extend through the thickness of tegument and have a free cilium inside a pit (types I and II). Two types (III and IV) are nonciliate and entirely intra-tegumental in location. Type IV receptor with large horizontal and thin vertical rootlets was described earlier in aspidogastreans only. Below the basal lamina, nerve endings in close association with muscle fibres, comparable with those in the Aspidogastrea, were detected. PMID- 14535349 TI - Some nematodes of fishes from central China, with the redescription of Procamallanus (Spirocamallanus)fulvidraconis (Camallanidae). AB - The present paper comprises a systematic survey of nematodes based on helminthological examinations of 176 specimens of freshwater fishes, belonging to 22 species, from central China (mostly lakes in Hubei Province) collected during the autumn of 2001. The following six species were recorded: Procamallanus (Spirocamallanus)fulvidraconis Li, 1935, Camallanus cotti Fujita, 1927, Dentiphilometra monopteri Moravec et Wang, 2002, Pingus sinensis Hsu, 1933, Proleptinae gen. sp. larv., and Eustrongylides sp. larv. Data on their morphology, morphological variability, host range, prevalence, intensity and distribution are provided. SEM studies of P. fulvidraconis and larval Physalopterinae, used for the first time in these species, revealed some additional morphological details and made it possible to redescribe the former. In contrast to the existing description of P. fulvidraconis, this species was found to possess two spicules and a V-shaped gubernaculum with unequal arms (originally mistaken for the left spicule), as well as deirids, whose location can be considered an important taxonomic feature. Larvae of the Physalopterinae have not previously been reported from fishes in China. The finding of larval Eustrongylides in Paramisgurnus dabryanus represents a new host record. All but one nematode species from this zoogeographically interesting region are briefly described and illustrated. PMID- 14535350 TI - Ecological analyses of the intestinal helminth communities of the wolf, Canis lupus, in Spain. AB - This work describes the ecological characteristics of the intestinal helminth communities of 50 wolves (Canis lupus L.) from Spain. The species found were classified into three groups according to prevalence, intensity and intestinal distribution. Taenia hydatigena Pallas, 1766 and Uncinaria stenocephala (Railliet, 1884) are the core species of the community. Taenia multiceps (Leske, 1780) is a secondary species. The rest of the species, Alaria alata (Goeze, 1782), Taenia serialis (Gervais, 1847). Taenia pisiformis (Bloch, 1780), Dipylidium caninum (Linnaeus, 1758), Mesocestoides sp. aff. litteratus, Toxocara canis (Werner, 1782), Toxascaris leonina (von Linstow, 1902), Ancylostoma caninum (Ercolani, 1859) and Trichuris vulpis (Froelich, 1789), behave as satellite species. The linear intestinal distribution of all helminth species was analysed. The location of most species can be considered predictable, especially for core and secondary species. The analysis of interspecific relationships between infracommunities shows that negative associations are more numerous than positive associations. The role of A. caninum in the community is compared with that of U. stenocephala. PMID- 14535351 TI - Babesia microti: parasitaemia and antibody responses to primary and challenge infections in BALB/c mice. PMID- 14535352 TI - Entrainment of circadian programs. AB - Of the three defining properties of circadian rhythmicity--persisting free running rhythm, temperature compensation, and entrainment--the last is often poorly understood by many chronobiologists. This paper gives an overview of entrainment. Where have we been? Where are we now? Whence should we be going? Particular emphasis is given to a discussion of the Discrete vs. Continuous models for entrainment. We provide an integrated mechanism for entrainment from a limit-cycle perspective. PMID- 14535353 TI - The circadian control of eclosion. AB - Eclosion is the stage in development when the adult insect emerges from the shell of its old cuticle. The sequence of behaviors necessary for eclosion is coordinated by an integrated system of hormones and is activated by hormones that relay developmental readiness. The circadian clock, which controls the timing of behaviors such as the rest: activity rhythm of adult insects, also controls eclosion timing. A number of groups are actively investigating the mechanisms by which the circadian clock restricts or gates eclosion to a particular time of day. Data from these studies are beginning to reveal details of the molecular and physiological basis of the eclosion rhythm. PMID- 14535355 TI - Effects of light and temperature on the circadian system controlling sperm release in moth Spodoptera littoralis. AB - Reproductive physiology of male moths is regulated by a peripheral circadian system, which controls the timing of sperm release from the testis into the upper vas deferens (UVD) and timing of sperm transfer from the UVD to the seminal vesicles. We investigated various effects of light and temperature on sperm release and transfer rhythms in the moth Spodoptera littoralis. We report that both rhythms persist for up to 1 week in constant darkness without significant dampening and are also temperature compensated in the range from 20 degrees C to 30 degrees C. However, the duration of sperm retention in the UVD is temperature dependent; consequently, temperature exerts a masking effect on the rhythm of sperm transfer. Experimental manipulations of light and temperature regime demonstrated that light dominates over temperature in entraining the timing of sperm release and transfer. Nevertheless, temperature plays a critical role in the absence of light Zeitgeber. Sperm release and transfer are arrhythmic in constant light (LL); however, both rhythms are restored by temperature cycles. PMID- 14535354 TI - Circadian aspects of postprandial metabolism. AB - Time-dependent variations in the hormonal and metabolic responses to food are of importance to human health, as postprandial metabolic responses have been implicated as risk factors in a number of major diseases, including cardiovascular disease. Early work reported decreasing glucose tolerance in the evening and at night with evidence for insulin resistance at night. Subsequently an endogenous circadian component, assessed in constant routine (CR), as well as an influence of sleep time, was described for glucose and insulin. Plasma triacylglycerol (TAG), the major lipid component of dietary fat circulating after a meal, also appears to be influenced by both the circadian clock and sleep time with higher levels during biological night (defined as the time between the onset and offset of melatonin secretion) despite identical hourly nutrient intake. These time-dependent differences in postprandial responses have implications for shiftworkers. In the case of an unadapted night shift worker, meals during work time will be taken during biological night. In simulated night shift conditions the TAG response to a standard meal, preceded by either a low-fat or a high-fat premeal, was higher after a nighttime meal than during a daytime meal, and the day/night difference was larger in men than in women. In real night shift workers in Antarctica, insulin, glucose, and TAG all showed an increased response after a nighttime meal (second day of night shift) compared to a daytime meal. Night shift workers are reported to have an approximately 1.5 times higher incidence of heart disease risk and also demonstrate higher TAG levels compared with matched dayworkers. As both insulin resistance and elevated circulating TAG are independent risk factors for heart disease, it is possible that meals at night may contribute to this risk. PMID- 14535356 TI - Daily profiles of plasma prolactin (PRL), growth hormone (GH), insulin-like growth factor-1 (IGF-1), luteinizing hormone (LH), testosterone, and melatonin, and of pituitary PRL mRNA and GH mRNA in male Long Evans rats in acute phase of adjuvant arthritis. AB - We studied the effects of adjuvant arthritis (AA) on the endocrine circadian rhythms of plasma prolactin (PRL), growth hormone (GH), insulin-like growth factor-1 (IGF-1), luteinizing hormone (LH), testosterone, and melatonin and of pituitary PRL and GH mRNA in male Long Evans rats. Groups of control and AA rats (studied 23 days after AA induction) that were housed under a 12/12 h light/dark cycle (light on at 06:00 h) were killed at 4 h intervals starting at 14:00 h. Cosinor analysis revealed a significant 12 h rhythm in PRL and PRL mRNA (p < 0.001) in controls with peaks at 14:00 h and 02:00 h, respectively. The peak at 02:00 h was abolished in the AA group resulting in a significant 24 h rhythm in parallel with that of PRL (p < 0.05) and PRL mRNA (p < 0.0001). Growth hormone showed no rhythm, but a significant rhythm of GH mRNA was present in both groups (p < 0.0001). Insulin-like growth factor-1 showed a 24 h rhythm in control but not in AA rats. The mean values of GH, GH mRNA, and IGF-1 were significantly reduced in AA. Luteinizing hormone displayed a significant 24 h rhythm (p < 0.01) peaking in the dark period in the control but not AA group. Testosterone showed in phase temporal changes of LH levels with AA abolishing the 02:00 h peak. Melatonin exhibited a significant 24 h rhythm in control (p < 0.001) and AA (p < 0.01) rats with maximum levels during the dark phase; the mesor value was higher in the AA males. These results demonstrate that AA interferes with the rhythms of all the studied hormones except the non-24 h (arrhythmic) GH secretion pattern and the rhythm in melatonin. The persistence of a distinct melatonin rhythm in AA suggests the observed disturbances of hormonal rhythms in this condition do not occur at the level of the pineal gland. PMID- 14535357 TI - Zeitgeber hierarchy in humans: resetting the circadian phase positions of blind people using melatonin. AB - Four blind individuals who were thought to be entrained at an abnormal circadian phase position were reset to a more normal phase using exogenous melatonin administration. In one instance, circadian phase was shifted later. A fifth subject who was thought to be entrained was monitored over four years and eventually was shown to have a circadian period different from 24 h. These findings have implications for treating circadian phase abnormalities in the blind, for distinguishing between abnormally entrained and free-running blind individuals, and for informing the debate over zeitgeber hierarchy in humans. PMID- 14535358 TI - Effect of evening exposure to dim or bright light on the digestion of carbohydrate in the supper meal. AB - In a previous study we found that daytime exposure to bright as compared to dim light exerted a beneficial effect on the digestion of the evening meal. This finding prompted us to examine whether the digestion of the evening meal is also affected by evening light intensity. Subjects lived in light of 200 lux during the daytime (08:00-17:00 h) and took their evening meal at 17:00 h under 20 lux (evening dim-light condition: 17:00-02:00 h) or 2000 lux (evening bright-light condition: 17:00-02:00 h) until retiring at 02:00 h. Assessment of carbohydrate digestion of the evening meal was accomplished by a breath hydrogen test that is indicative of the malabsorption of dietary carbohydrate. Hydrogen excretion in the breath in the evening under the dim-light condition was significantly less than under the bright-light condition (p < 0.05). This finding is the opposite to that obtained in previous experiments in which subjects were exposed to the different intensities of light during the daytime, and indicates that the exposure to dim light in the evening exerts a better effect on carbohydrate digestion in the evening meal than does the exposure to bright light. PMID- 14535359 TI - Circadian variation in cortisol reactivity to an acute stressor. AB - To investigate the role of the circadian pacemaker in cortisol reactivity to a cold pressor challenge, 26 diurnally subjects participated in a constant-routine protocol and were divided into two groups. Group 1 started immediately after a monitored sleep period at 09:00 h, while group 2 started 12 h later. After 2 h of adaptation, a cold pressor test was presented every 3 h. The cortisol response was assessed by means of saliva samples that were taken before and after the test. The pretest samples were considered to be base-rate measures and base-rate values as subtracted from post-test values were considered as reactivity measures. Both measures showed distinct Time-of-Day variations (respectively: F(7,168) = 16.92, p < 0.001, epsilon = 0.383; and F(7,175) = 8.01, p < 0.001, epsilon = 0.523). These findings are interpreted as evidence for the existence of an endogenous circadian periodicity underlying the sensitivity of the hypothalamus-pituitary-adrenal (HPA)-axis to acute stress. PMID- 14535360 TI - Diurnal variations in cycling kinematics. AB - Physiological and biomechanical constraints as well as their fluctuations throughout the day must be considered when studying determinant factors in the preferred pedaling rate of elite cyclists. The aim of this study was to monitor the diurnal variation of spontaneous pedaling rate and movement kinematics over the crank cycle. Twelve male competitive cyclists performed a submaximal exercise on a cycle ergometer for 15 min at 50% of their W(max). Two test sessions were performed at 06:00 and 18:00 h on two separate days to assess diurnal variation in the study variables. For each test session, the exercise bout was divided into three equivalent 5-min periods during which subjects were requested to use different pedal rates (spontaneous cadence, 70 and 90 rev min(-1)). Pedal rate and kinematics data (instantaneous pedal velocity and angle of the ankle) were collected. The results show a higher spontaneous pedal rate in the late afternoon than in the early morning (p < 0.001). For a given pedal rate condition, there was a less variation in pedal velocity during a crank cycle in the morning than in the late afternoon. Moreover, diurnal variations were observed in ankle mobility across the crank cycle, the mean plantar flexion observed throughout the crank cycle being greater in the 18:00 h test session (p < 0.001). These results suggest that muscular activation patterns during a cyclical movement could be under the influence of circadian fluctuations. PMID- 14535361 TI - Case study: differences in human Per2 gene expression, body temperature, cortisol, and melatonin parameters in remission and hypersomnia in a patient with recurrent hypersomnia. AB - Recurrent hypersomnia is characterized by recurring episodes of hypersomnia of 18 h or more per day lasting from several days to several weeks. We report the case of a 17-year-old male subject with recurrent hypersomnia who displayed change in the 24 h expression of the hPer2 gene in whole red and white blood cells as well as markers [deep body temperature (DBT) and cortisol] of the circadian time structure during an episode of hypersomnia compared to remission. The patient was studied for the temporal characteristics of hPer2 gene, DBT, cortisol, and melatonin expression during a single 24 h span during an episode of hypersomnia and again during a single 24 h span in the following remission. The approximation of a 24 h cosine curve to the time series data revealed circadian rhythmicity (P < 0.05) only in DBT in the two stages of the disease with differences in amplitude and acrophase. Cortisol circadian rhythmicity was detected during remission, but not during hypersomnia. Statistically significant differences were detected by ANOVA between the remission and active disease stages in the 24 h mean level of hPer2 gene expression (P < 0.05), cortisol (P < 0.05), and DBT (P < 0.05). The findings of this case study suggest the expression of hPer2 gene and alterations in circadian time structure might play an important role in the pathogenesis of recurrent hypersomnia, although additional study is required. PMID- 14535362 TI - Variant Creutzfeldt-Jakob disease. PMID- 14535363 TI - BSE risks for humans consuming beef and beef products: how any risks are managed. PMID- 14535364 TI - Pathogenesis of BSE. AB - Before the emergence of bovine spongiform encephalopathy (BSE) and recognition of its zoonotic potential, the major example of the transmissible spongiform encephalopathies (TSEs) of animals was scrapie of sheep. But there is no evidence that scrapie transmits naturally to any species other than sheep and goats. The pathogenesis of scrapie has been studied most in experimental laboratory rodent species. In most experimental models of scrapie, after peripheral non-neural routes of infection, replication of the agent can first be detected in lymphoreticular system (LRS) tissue. When the route of introduction of agent into the body is localized, initial involvement will be in LRS tissue draining the infection site. Thereafter, there is a striking amplification of the agent in the LRS and spread by lymphatic/haematogenous routes, giving widespread dissemination in the LRS. This precedes replication in the CNS, but is not the means by which infection reaches the CNS. There is now substantial evidence from experimental models of scrapie that involvement of the CNS is by peripheral nervous system (PNS) pathways. In some models employing oral exposure the earliest localized LRS replication is in the gut-associated lymphoid tissue (GALT) and autonomic PNS routing to the CNS has been implicated. However, the relative importance of different routes of spread of TSEs within the body is determined by a number of host- and agent-dependent factors and, therefore, generalizations from an experimental model to a natural disease across a species barrier may not be appropriate. With the occurrence of BSE and recognition of its food-borne route of transmission via meat and bone meal, has come greater awareness of the probable importance of the oral route of infection in ruminant species affected by TSEs. In consequence, studies have increasingly focused on the natural host species to examine pathogenetic events. PMID- 14535365 TI - The role of CEA (Center of Animal Encephalopathies) in the BSE surveillance: BSE in Italy. PMID- 14535366 TI - Animal transmissible spongiform encephalopathies and genetics. AB - The genotype of the host plays a crucial role in the pathogenesis of transmissible spongiform encephalopathies (TSEs). In this respect, the most important factor is represented by the gene of the prion protein (PrP). The present work summarizes the currently available knowledge on the genetic basis of TSEs focusing, in particular, on sheep scrapie. Interest in this disease has grown markedly following the discovery of bovine spongiform encephalopathy, both for scientific and health reasons. In Italy, specific research grants from the Ministry of Health and the National Research Council (CNR), together with cooperation between the Istituto Superiore di Sanita and the Istituti Zooprofilattici Sperimentali, have allowed us to study the PrP genotype and to investigate the genetic susceptibility to scrapie in the most important Italian sheep breeds, with special reference to Sarda, Comisana and Massese. The PrP genotype in relation to scrapie susceptibility was also studied in goats of Ionica breed. PMID- 14535367 TI - Impact of BSE on livestock production system. AB - The small number of BSE cases diagnosed in Italy from January 2001 to 12 September 2001 (a total of 28, one every 9000 head) does not allow for a statistical analysis of the relationship between this disease and the livestock systems. However, some indications can be noted: (a) only dairy cattle, which represent three-quarters of the cattle raised in Italy, are involved; (b) 58% of the cases belong to medium-large farms that breed 27% of all head; (c) 13 out of 28 cases are 5-year-old animals and 26 out of 28 are between 5 and 7 years of age; (d) 15 of 28 cases come from Lombardia, where 27% of Italian dairy cattle are raised. The following factors may have affected the livestock system: (1) trends of beef meat consumption; (2) changes in livestock management; (3) changes in animal feeding; (4) possible effects on selection. A strong decline in beef meat consumption (4 kg/year) has been observed in the UK and other European countries since 1996 (the year of the discovery of the relationship between BSE and nvCJD). In Italy, from January 2001 the consumption of beef meat has declined as well as slaughter: a drop of 31% in the total slaughtered head in the period January-February, a drop of 14% in January-May. A fall in the price of calves has promoted, in some dairy farms, the start of the production of light beef less than one year old (advantages in the marketing of meat favour this initiative), a phenomenon which is not yet well established. Traceability and certification of meat have improved, thanks to breeders' associations and interprofessional agreements. The breeders associations have also started insurance initiatives against BSE risks. In Italy the employment of plant protein meals would increase the total feedstuff consumption by about 7%. Direct effects of BSE could slow down the genetic progress (GP) of cattle populations within breed and country. Indirect effects on GP may also happen as a consequence of an increase in the replacement rate (rr). This increase in rr reduced the generation interval and will therefore proportionally increase GP. Some important questions for the livestock production system are: Does the vertical transmission of BSE exist? Is there a genetic basis favouring the disease and is it inheritable? Are in vivo diagnostic tests possible? Are vaccination schemes against BSE possible and useful or is it better to pursue eradication? PMID- 14535368 TI - Critical observations on BSE control in Italy. PMID- 14535369 TI - The 'BSE Strategic Project' of the National Council of Research: results of four years of research. PMID- 14535370 TI - The BSE emergency in Lombardy. PMID- 14535371 TI - The anatomy and physiology of diarthroses. PMID- 14535372 TI - Pathophysiology of osteoarthritis. PMID- 14535373 TI - Arthroscopy in the horse. PMID- 14535374 TI - Arthroscopy in the dog. PMID- 14535375 TI - Swine influenza in Italy. PMID- 14535376 TI - Ecological aspects of influenza A virus circulation in wild birds of the Western Palearctic. PMID- 14535377 TI - Long-term monitoring for avian influenza viruses in wild bird species in Italy. PMID- 14535378 TI - Influenza viruses: structure and interspecies transmission mechanisms. PMID- 14535379 TI - The 1999-2000 avian influenza (H7N1) epidemic in Italy. PMID- 14535380 TI - Physiological and metabolic adaptations in the mammary gland and consequences for the dairy cow. PMID- 14535381 TI - Transition cow: non-specific immune response. PMID- 14535382 TI - Transition cow: interaction with fertility. AB - In recent years a progressive worsening of fertility indices in dairy cow herds has been observed. Several factors (genetic, dietary and management) seem to be more related to poor fertility than milk yield level. The degree and the length of the energy deficit during the transition period are inversely related to reproductive indices (e.g. conception rate is <30% for BCS decreases over one unit). A serious energy deficit reduces (or suppresses) pulsatile secretion of gonadotrophins (ovarian dysfunction and/or smaller follicles): IGF-I and insulin plasma levels (slower follicle growth and higher embryonic mortality); and progesterone production from the corpus luteum (higher rate of embryonic abortions). The diet influences fertility in several ways. Excess of rumen degradable proteins, apart from negative energy balance, negatively affects reproductive activity. Conversely, some nutrients (i.e. some polyunsaturated fatty acids or some amino acids) seem to show positive effects on fertility. Finally, the relationship between health status, often compromised during the transition period, and fertility efficiency is discussed. The release of cytokines seems to be related directly and indirectly (mainly by the change in usual hepatic metabolism to the malfunction of reproductive apparatus. Quick recovery of reproductive activity requires the adoption of strategies around calving to cover the higher environmental and nutritive requirements and to prevent disorders of any kind. PMID- 14535383 TI - Transition cow: nutritional prophylaxis. PMID- 14535384 TI - Dioxin and PCB contamination of fish and shellfish: assessment of human exposure. Review of the international situation. PMID- 14535385 TI - Analysis of the risks in the marketing of fishing products. PMID- 14535386 TI - Algal biotoxins of marine origin: new indications from the European Union. AB - Marine biotoxins, more or less complex molecules with various origins that can accumulate in the tissues of fish products through the food chain, are reviewed. The EU, aware of the danger incurred in eating certain fish products, has issued a set of hygiene and health directives for the purpose of preventing disease and safeguarding consumer health. In particular, directive 91/492/EEC, of 15 July 1991, lays down the sanitary norms applicable to the production and commercialization of live bivalve molluscs, echinoderms, tunicates and marine gastropods and regulates the whole system involving these products from their origin to consumption. More recently, through Commission Decision dated 15 March 2002 (EC OJ 175/62 of 16.3.2002) the EU has set new standards for the implementation of directive 91/492/EEC with respect to the maximum levels and analysis methods for some marine biotoxins. PMID- 14535388 TI - Sex determination in the common buzzard (Buteo buteo) by means of endoscopy and DNA analysis. PMID- 14535389 TI - Effect of physical exercise on thiols in the plasma in the athletic horse: connection with the immune system. PMID- 14535387 TI - Heavy metal, polychlorinated biphenyl and organochlorine pesticide residues in marine organisms: risk evaluation for consumers. AB - This survey provides information on the levels of heavy metal, polychlorinated biphenyl and organochlorine pesticide residues in marine organisms to ascertain whether these concentrations exceeded the prescribed legal limits. In order to assess the potential human health impact, the weekly intake was estimated. Most of the organisms analysed showed higher levels of mercury than the maximum permissible limit, while cadmium and lead were below the proposed permissible limits in all samples. The estimated intake was far above the established Provisional Tolerable Weekly Intake for all metals, except for total mercury. PMID- 14535390 TI - Apoptosis in cooled porcine oocytes: role of calcium (Ca2+) and Ca2+-dependent enzymes. PMID- 14535392 TI - Evaluations of testicular biopsy by tru-cut in the stallion. PMID- 14535391 TI - Na,K-ATPase and other parameters in bivalve molluscs from the Adriatic Sea under different environmental conditions. PMID- 14535393 TI - Biochemical changes during post-larval growth in white muscle of gilthead sea bream (Sparus aurata) fed zinc-fortified diets. PMID- 14535394 TI - Effect of food deprivation on blood concentration of metabolic hormones in Merino rams: the role of leptin. PMID- 14535395 TI - Endothelin-1 (ET-1) involvement in respiratory dysfunctions during endotoxic shock in the pig. PMID- 14535396 TI - Apoptosis and expression of related proteins in mammary gland of heifers during early lactation. PMID- 14535397 TI - Variations in some electrocardiographic parameters in the trotter during racing and training. PMID- 14535398 TI - VEGF, bFGF and swine granulosa cells: proliferation, steroidogenesis and NO production. PMID- 14535399 TI - Alpha-inhibin expression in canine ovarian neoplasms: preliminary results. PMID- 14535401 TI - Relationship between dysplastic and neoplastic mammary lesions and pseudopregnancy in the bitch. PMID- 14535400 TI - Paternal chromatin remodelling in mouse oocytes following fertilization. PMID- 14535402 TI - Estimation of the content of pectins in feeds: fractional extraction and quantitative determination. PMID- 14535404 TI - Estimation of dry matter intake by grazing Comisana ewes. PMID- 14535403 TI - Seasonal variations of free fatty acids in plasma of 'Nero Siciliano' pigs living in extensive conditions. PMID- 14535405 TI - Different protein sources in piglet feeding. PMID- 14535406 TI - Milk choline, alpha-tocopherol and neutrophil chemotaxis in the periparturient dairy cow. PMID- 14535407 TI - Composition, acidity and rennet-coagulation properties of early- and late lactation milks from Italian Friesian cows. PMID- 14535408 TI - Effect of the use of maize-silage with low aerobic stability on the performance of lactating dairy cows. PMID- 14535409 TI - The standardization of a molecular biology method to verify the presence of Microcystis aeruginosa. PMID- 14535410 TI - Isolation of Salmonella enterica from slaughtered pigs. PMID- 14535411 TI - Trawling lesions: incidence in some fish species and preliminary statistical evaluations. PMID- 14535412 TI - Typing of Staphylococcus aureus isolates from raw milk. PMID- 14535413 TI - Effect of NaCl concentration on the survival and growth of coliforms in raw seasoned sausages. PMID- 14535414 TI - Cd, Hg and As concentrations in fish caught in the North Adriatic Sea. PMID- 14535415 TI - Inspective consideration in detection of enterobacteria in the blood of normally slaughtered swine. PMID- 14535416 TI - Isolation of cytotoxic Aeromonas spp. from food. PMID- 14535417 TI - Low-cost biosensors: an inspective application in milk control. PMID- 14535418 TI - Occurrence of lesions in the organs of regularly slaughtered cattle. PMID- 14535419 TI - Pseudotuberculosis in sheep as a concern of veterinary public health. PMID- 14535420 TI - Behaviour of Listeria monocytogenes in anchovies during marination. PMID- 14535421 TI - Effect of light exposure on lipid and colour stability of Southern and Northern European type sausages. PMID- 14535422 TI - First report of morphological variants of urothelial carcinoma in the dog. PMID- 14535423 TI - Transmissible spongiform encephalopathies: in-vitro evaluation of the therapeutic potentiality of new molecules. PMID- 14535424 TI - Application of biomolecular methods to Staphylococcus aureus strains from dairy cows. PMID- 14535425 TI - Use of degenerate PCR to identify an anti-apoptotic gene of BHV-1 corresponding to the gamma34.5 gene of HSV-1. PMID- 14535426 TI - Expression of the focal adhesion protein paxillin in normal and breast cancer tissues. PMID- 14535427 TI - Development of in vitro cell cultures for the evaluation of molecules with antiprionic activity. PMID- 14535428 TI - Isolation stress in sheep: effects on neutrophil gene expression of CD18, IL8 and C5a receptors. PMID- 14535430 TI - Canine ovarian tumours: a retrospective study of 49 cases. PMID- 14535429 TI - Molecular analysis of the NP gene of Italian CDV isolates. PMID- 14535431 TI - Endogenous retroviruses in swine cell lines and evaluation of possible transmission to primate cellular systems. PMID- 14535432 TI - Acute myeloid leukaemia in five dogs: clinical findings and cytochemical characterization. PMID- 14535433 TI - Doppler evaluation of the ophthalmic vasculature in the beagle: normal values. PMID- 14535434 TI - Effects of the administration of spiramycin and tiamulin on in vivo complex formation and hepatic cytochrome P4503A (CYP3A)-dependent monooxygenases in rabbits. PMID- 14535435 TI - Use of computed tomography in thoracic diseases of small animals. PMID- 14535437 TI - Hypertrophic obstructive cardiomyopathy associated to mitral valve dysplasia in the Dalmatian dog: two cases. PMID- 14535436 TI - Medroxyprogesterone acetate increases anthracycline-induced lipid peroxidation and membrane fluidity. PMID- 14535438 TI - Treadmill endoscopic findings in 15 racehorses presented for poor performance. PMID- 14535439 TI - Evaluation of the clinical signs and computed tomographic findings in 27 dogs with intracranial space-occupying lesions (1999-2000). PMID- 14535440 TI - A case report of T-cell lymphoma in a horse. PMID- 14535441 TI - Clinical usefulness of peripheral blood lymphocyte subsets in canine lymphoma. PMID- 14535442 TI - Peritoneal ultrasonography in dogs and cats. PMID- 14535443 TI - Identification of functional beta-adrenoceptors in Caco-2 cell membranes. PMID- 14535444 TI - Emphysematous cystitis in two glycosuric dogs. PMID- 14535445 TI - Evaluation of leukotriene B4 in the canine exhaled breath: standardization of a technique of sample collection. PMID- 14535446 TI - Effects of interferon alpha (INF-alpha) therapy on peripheral blood lymphocyte subsets from FIV and FeLV naturally infected cats. PMID- 14535447 TI - Kinetics of ketamine plasma and urine metabolite levels following intravenous administration in the dog. PMID- 14535448 TI - Chronic renal failure (CRF) in horses: personal reports. PMID- 14535449 TI - Sebaceous adenitis in the dog: three cases. PMID- 14535450 TI - Application of the 'extraction bag' in laparoscopic treatment of unilateral and bilateral abdominal cryptorchidism in dogs. PMID- 14535451 TI - Diagnostic and prognostic value of serum lactate dehydrogenase (LDH) and LDH isoenzymes in canine lymphoma. PMID- 14535452 TI - Short and long-term prospects for consumption of fish. AB - If real prices for fish remain at the levels they had attained by 1999, in the year 2050 demand for fish and shellfish as food could be of the order of 270 million tons (live weight equivalent) per year. If producers were able to supply these quantities consumption would rise by 176% over this 50-year period. To meet the demand supply would have to expand at the rate of 2.1% annually; but, a review of the pattern of population growth--and of historical patterns of increases in per capita consumption of fish--shows that annual growth in the volume of fish demanded is likely to be largest in the coming two decades, and then to taper off. Will producers be able to deliver? It is clear that wild marine stocks at present harvested by capture fishermen cannot support fisheries that would yield much more than 100 million tons per year and of this amount a significant proportion will continue to be used for fish meal and oil production. The question therefore narrows down to: can aquaculture, or non-traditional marine species, supply the required amounts? The historical context of supply is considerably different from that which has prevailed during the past 30 years. At that time the growing demand in OECD countries was met partly through imports of fish produced in the seas and lakes of developing countries. During coming decades the increased demand in developing countries must be met essentially through their own resources. In fact, in poor countries it seems unlikely that supply will respond to demand unless they experience economic growth. PMID- 14535453 TI - Aquaculture and disease control in Italy. PMID- 14535455 TI - Species identification and safety of fish products. PMID- 14535454 TI - Emerging pathologies in aquaculture: effects on production and food safety. AB - Infectious diseases represent a limiting factor for the further development of Italian aquaculture. The recent introduction and spreading of new pathogens, along with the global climatic change, has contributed to a considerable decrease in trout production. Emerging pathologies in rainbow trout culture include viral diseases, e.g. infectious haematopoietic necrosis (IHN), bacterial diseases, such as lactococcosis and visceral flavobacteriosis, and parasitical diseases, e.g. proliferative kidney disease (PKD). Higher mortality rates in trout fry and fingerlings are generally induced by visceral flavobacteriosis and IHN, while the main losses in large trout during the warm season are due to lactococcosis and PKD. Mariculture has at present a better sanitary status compared to trout culture, but a rapid dissemination of pathogens, including zoonosis agents, is envisaged also for seabass and seabream. Emerging pathologies in sea bass include VNN, pseudotuberculosis, streptococcosis and tuberculosis. Seabream is much more resistant and is mainly affected by novel Vibrio infections and enteromyxidiosis. A good sanitary management of fish farms is essential for avoiding or limiting losses caused by emerging pathologies. Transmission of zoonosis agents to man, through the consumption of cultured fish, is very remote in Italy. On the contrary, transmission of Streptococcus iniae, Vibrio vulnificus and Mycobacterium marinum by means of improper manipulation of infected fish, could represent a potential hazard for fish farmers and fish processors, as well as for people preparing fish meals. PMID- 14535456 TI - Application of risk management to the production chain of intensively reared fish. AB - Principles of risk management, as a part of risk analysis, are described with respect to their application to the production chain of intensively reared fish. According to the outcomes of the FAO/WHO Expert consultation on the Application of Risk Management (Rome, 1997), there are four steps: risk evaluation, risk management option assessment, implementation of management decisions, and monitoring and review. There is a lack of information on risk assessment for farmed fish though, regarding bacteriological hazards, the data on farmed fish pathology and microbiology suggest a potential prevalence of Vibrio spp. and Aeromonas spp. Other bacterial contaminants could be introduced on handling during selection and packaging of products. On the basis of the above qualitative risk evaluation, risk management options have been individuated. These concern the monitoring of the health status of fish during the fattening period, harvest parameters, hygiene of selection and packaging, and storage characteristics. PMID- 14535457 TI - Traceability issues in fishery and aquaculture products. PMID- 14535458 TI - The health benefit of seafood. PMID- 14535459 TI - Large animals transportation procedures in Europe: present and future. PMID- 14535460 TI - Causes of poor welfare in large animals during transport. AB - The welfare of animals during transport should be assessed using a range of behavioural, physiological and carcass quality measures. In addition, health is an important part of welfare so the extent of any disease, injury or mortality resulting from, or exacerbated by, transport should be measured. Many of the indicators are measures of stress in that they involve long-term adverse effects on the individual. Key factors affecting the welfare of animals during handling and transport which are discussed are: attitudes to animals and the need for training of staff; methods of payment of staff; laws and retailers' codes; genetics, especially selection for high productivity; rearing conditions and experience; the mixing of animals from different social groups; handling procedures: driving methods; stocking density; increased susceptibility to disease and increased spread of disease. PMID- 14535462 TI - Effects of transport on health of farm animals. PMID- 14535461 TI - Evaluation of stress during transport. AB - Domestic animals are transported for a variety of reasons including breeding, biomedical purposes, slaughter and, in the case of sporting horses, for competitions, pleasure activities or ceremonial proceedings. Studies to determine the amount of stress on farm animals during transport often have highly variable results and are difficult to interpret. The reaction of animals to stressors depends on the duration and intensity of the stressors, the animal's previous experience, its physiological status and the immediate environmental restraints. Behavioural, haematological, haematochemical, physiological and neuro-hormonal (beta-endorphin, ACTH, cortisol, iodothyronines) variables are discussed on the basis of handling, loading and transport procedures of animals. PMID- 14535463 TI - The transport of animals in the European Union: the legislation, its enforcement and future evolutions. AB - Animal transport is an important activity of the farming industry. It is also the most controversial area of animal welfare. After presenting the current legislative framework in the European Union that protect animals during transport and the main enforcement difficulties, this paper considers the international, political and scientific perspectives that lead the European Commission to draft new rules in this regard. PMID- 14535464 TI - Immune system and ageing in the dog: possible consequences and control strategies. PMID- 14535465 TI - Diagnosis and management of geriatric canine endocrine disorders. PMID- 14535466 TI - Cardiovascular diseases in the ageing dog: diagnostic and therapeutic problems. PMID- 14535467 TI - Workshop on wildlife: conservation and sanitary aspects. PMID- 14535468 TI - General directives for wildlife protection. PMID- 14535469 TI - Wildlife veterinarian, conservation and public health. AB - In recent years wildlife diseases (infectious and non-infectious) have played a relevant role in both wildlife conservation and public health. Global environmental changes have determined a bimodal evolution of wildlife. On one side a huge loss of biodiversity has been observed leading to the increasing of threatened or endangered species. In contrast few opportunistic taxa increased their aboundances and ranges. The above scenarios claim the intervention of wildlife veterinarians. In conservation the understanding of the ecological role of the host parasite relationship and the perturbations on the host population dynamics have to be assessed and eventually modified. In public health the increased overlapping among wildlife, livestock, pets and human beings represents a risk for diseases spread (no matter in which directions). Serious limits are, still now, observed in the acceptance of this 'new world' by veterinary academics. As a consequence curricula often fail in providing adequate skill at both undergraduate and graduate levels. An addressed approach towards wildlife diseases should be promoted as an essential component of environmental management. PMID- 14535470 TI - Alteration of constitutive heat shock protein 70 (HSC70) production by in vitro culture of porcine preimplanted embryos. PMID- 14535471 TI - Central distribution of A delta and C afferent fibres from the dental pulp and periodontium in the rat. PMID- 14535472 TI - In vitro growth of sheep preantral follicles: which perspectives? PMID- 14535473 TI - Evaluation of alpha-glucosidase activity in dog semen and its use in fertility diagnosis. PMID- 14535474 TI - Presence of S100-like protein in non-mammalian vertebrate kidney. An immunocytochemical study. PMID- 14535475 TI - Periodic monitoring of some physiological parameters during training in the athletic horse. PMID- 14535477 TI - Double labelling immunohistochemistry on the peripheral autonomic neurons projecting to the bulbospongiosus muscle in male impuberal pigs. PMID- 14535476 TI - Characterization of serum 'glycosaminoglycan (GAG) chains' and 'native proteoglycan' fractions in the ostrich. PMID- 14535478 TI - Mare embryonic resorption and homocysteine. PMID- 14535480 TI - Effect of manual imitation of grooming on riding horses' heart rate in different environmental situations. PMID- 14535479 TI - Expression of inhibin/activin alpha, betaA and betaB subunits in the ovary of the bitch. PMID- 14535481 TI - Embryo recovery rate in a mare affected by Cushing's syndrome. PMID- 14535482 TI - Localization of the telomerase catalytic subunit (TERT) in pig ovarian follicles. PMID- 14535484 TI - Recovery rate and recovery time in cows with luteal and follicular ovarian cysts after treatment with buserelin. PMID- 14535483 TI - Biochemical analysis of the molecular mechanism of action of natural integrin inhibitors. PMID- 14535485 TI - Endonuclease activity in swine ovarian cysts. PMID- 14535486 TI - Preliminary studies on compatibility between high yield levels and the well-being of dairy cows. PMID- 14535487 TI - Effects of the dietary inclusion of vegetable fat and dehydrated alfalfa meal on the technological properties of rabbit meat. PMID- 14535488 TI - Effect of body condition score variation on milk yield and fertility in post partum dairy cows. PMID- 14535489 TI - Influence exerted by certain factors during rearing and before slaughter on post mortem characteristics of sea bass. PMID- 14535490 TI - Detection of cross-contamination in feedstuffs: presence of constituents of animal origin. PMID- 14535491 TI - Antigenic recombinant proteins expressed in tobacco seeds as a model for edible vaccines against swine oedema. PMID- 14535492 TI - Seasonal variations of milk characteristics and cheesemaking losses in the manufacture of Parmigiano-Reggiano cheese. PMID- 14535493 TI - Design of universal primers for the detection of animal tissues in feedstuff. PMID- 14535494 TI - Identification with multiplex PCR assay of enterococcus species isolated from dairy products in Umbria, Italy. PMID- 14535495 TI - Identification of moulds belonging to the Penicillium genus using a polymerase chain reaction (PCR-SSCP) method followed by 'computer assisted' statistical analysis of the electrophoretic patterns. PMID- 14535496 TI - A new multiplex PCR for the detection of hbl genes in strains of the 'Bacillus cereus group'. PMID- 14535497 TI - Physico-chemical and biochemical modifications of Mytilus galloprovincialis after harvesting. PMID- 14535498 TI - Immunological and histochemical investigation of darkened turkey muscles. PMID- 14535499 TI - Effect of copper exposure on the antioxidant enzymes in bivalve mollusc Scapharca inaequivalvis. PMID- 14535500 TI - Farmed fish pathology: quality aspects. PMID- 14535501 TI - Detection of genetically modified organisms (GMOs) in food and feedstuff. PMID- 14535502 TI - Characterization of endogenous retroviruses in pig cell cultures. PMID- 14535503 TI - Diagnostic and prognostic features of feline cutaneous mast cell tumours: a retrospective analysis of 40 cases. PMID- 14535504 TI - BSE: active and passive control system. Data comparison in the district of Lodi. PMID- 14535505 TI - Acquired canine myasthenia gravis associated with thymoma: histological features and immunohistochemical localization of HLA type II and IgG. PMID- 14535506 TI - Development of in vitro cell cultures for the evaluation of molecules with antiprionic activity. PMID- 14535507 TI - The effect of natural and pharmacological stressors on sheep: haematological, biochemical and granulocytic functional changes. PMID- 14535508 TI - Characteristic clinical, haematological and histopathological findings in feline mastocytoma. PMID- 14535509 TI - Electrophoresis using pretreatment of serum with neuraminidase and affinity electrophoresis in the evaluation of ALP isoenzymes. PMID- 14535510 TI - Complex congenital heart disease: prevalence and clinical findings. PMID- 14535511 TI - Diagnosis and treatment of progressive ethmoidal haematoma (PEH) in horses. PMID- 14535512 TI - Brainstem auditory evoked-potentials in dairy cows recorded with Nape-Vertex derivation. PMID- 14535513 TI - Endoscopic examination of equine upper respiratory tract by computed image analysis. PMID- 14535514 TI - Clinical, laboratory, ultrasonographic and histopathological findings in dogs affected by portosystemic shunts, following surgery or medical treatment. PMID- 14535516 TI - Clinical and clinicopathologic findings in three dogs with steroid-responsive meningitis-arteritis (SRMA). PMID- 14535515 TI - Gastroscopic findings in 63 Standardbred racehorses in training. PMID- 14535517 TI - Clinical findings associated with Borrelia burgdorferi infection in the dog. PMID- 14535518 TI - Canine leishmaniasis in the newborn puppy. PMID- 14535519 TI - Effect of storage and cryopreservation on the lymphocyte responses to polyclonal mitogens in cattle. PMID- 14535520 TI - Detection of proinflammatory and regulatory cytokines in bovine milk using RT PCR. PMID- 14535522 TI - Tumour thrombus: direct endoluminal 'caudal caval vein-right atrium' extension in a dog affected by adrenal neoplasia. PMID- 14535521 TI - Intermittent gastroesophageal intussusception in a dog: clinical features, radiographic and endoscopic findings, and surgical management. PMID- 14535523 TI - Inflammatory bowel disease mimicking alimentary lymphosarcoma in a cat. PMID- 14535524 TI - Feline idiopathic ulcerative dermatosis: three cases. PMID- 14535525 TI - Asymptomatic dilated cardiomyopathy in Great Danes: clinical, electrocardiographic, echocardiographic and echo-Doppler features. PMID- 14535526 TI - Intraoperative electroencephalographic monitoring: quantitative analysis of bioelectrical data detected during surgical stimulation. PMID- 14535527 TI - Cadmium as an environmental factor of hypertension in animals: new perspectives on mechanisms. PMID- 14535528 TI - The use of electrocardiographic recording with Holter monitoring during treadmill exercise to evaluate cardiac arrhythmias in racehorses. PMID- 14535529 TI - New approaches to breast cancer therapy. AB - Breast cancer treatment is a rapidly changing area of oncology, with advances in diagnosis, treatment and prevention reported each year. Endocrine therapy is particularly important in the management of the disease because of its demonstrated efficacy and favorable safety profile. In response to rapidly emerging data, it is therefore crucially important that the positioning of established endocrine treatments is continually reassessed relative to the newer agents, thereby allowing clinical practice to be adapted in a measured and informed manner. This reassessment should also ensure that the broad treatment options (chemotherapy, endocrine therapy, and biological therapy) are no longer seen as discrete entities, but rather viewed as being complementary, with each agent used at an appropriate point within a planned, individualized treatment schedule. In this way, the therapeutic options can be refined and adapted to the ultimate benefit of all women with breast cancer. PMID- 14535530 TI - Three years' follow-up from the ATAC trial is sufficient to change clinical practice: a debate. AB - Tamoxifen is currently the adjuvant treatment of choice for postmenopausal women with hormone-sensitive breast cancer. However, in the treatment of postmenopausal women with advanced disease, the third-generation aromatase inhibitor anastrozole ('Arimidex') has been shown to be at least as effective as tamoxifen, and to be more effective than tamoxifen in patients with estrogen receptor-positive disease. Furthermore, anastrozole is well tolerated and is associated with fewer adverse reactions (such as thromboembolic events, vaginal bleeding, and endometrial cancer) compared with tamoxifen. A change in clinical practice has now emerged for the first-line treatment of postmenopausal advanced disease in patients, with tamoxifen becoming the second- or third-line choice for many clinicians. These data have raised questions about the optimal adjuvant treatment for postmenopausal women with early breast cancer. The 'Arimidex', Tamoxifen, Alone or in Combination (ATAC) trial has compared the efficacy and safety of tamoxifen and anastrozole in the adjuvant treatment of postmenopausal women with early breast cancer. At 3 years' follow-up in the overall population, anastrozole demonstrated a significant benefit compared with tamoxifen for disease-free survival (DFS) (89.4% vs. 87.4%; p = 0.013), time to recurrence (hazard ratio = 0.79; p = 0.008), and contralateral breast cancers (odds ratio = 0.42; 95% confidence interval: 0.22-0.79; p = 0.007). Anastrozole produced improvements in quality of life similar to tamoxifen and was better tolerated for a number of predefined adverse events. Of course, a large body of evidence is available regarding the safety profile of tamoxifen and some feel that more data are needed from the ATAC trial to demonstrate that the early advantages of anastrozole over tamoxifen can be maintained in the longer term. However, a follow-up analysis at 47 months has confirmed that the tolerability profile and the absolute benefit of anastrozole were maintained over the extended follow-up period, demonstrating that the benefits of anastrozole are likely to be maintained over the long term. This review assesses these and other data from the ATAC trial and presents the arguments for and against whether 3 years' follow-up is sufficient to inform a change in clinical practice for the adjuvant treatment of postmenopausal women with early breast cancer. PMID- 14535531 TI - Sequential hormonal therapy for metastatic breast cancer after adjuvant tamoxifen or anastrozole. AB - The use of adjuvant endocrine therapy in the treatment of hormone receptor positive, early breast cancer has become important in both pre- and postmenopausal women. Tamoxifen has been the principal adjuvant hormonal therapy in pre- and postmenopausal women with hormone receptor-positive breast cancer for nearly 20 years. Recent data in premenopausal women suggest benefit from ovarian ablation with or without tamoxifen. Early results from the 'Arimidex', Tamoxifen, Alone or in Combination (ATAC) trial have demonstrated that the third-generation, selective aromatase inhibitor (AI) anastrozole ('Arimidex') is a suitable alternative adjuvant therapy for postmenopausal women with hormone receptor positive disease. After recurrence or relapse on adjuvant endocrine therapy, responses to the sequential use of additional endocrine agents are common. The increase in the number of options now available for adjuvant therapy will have important implications for the selection of the optimal sequence of endocrine agents in the treatment of recurrent breast cancer. Menopausal status is an important factor in determining the endocrine therapy that a patient receives. For premenopausal women, tamoxifen and/or a luteinizing hormone-releasing hormone agonist such as goserelin ('Zoladex') are both options for adjuvant endocrine treatment. After progression on adjuvant and first-line tamoxifen, ovarian ablation is an appropriate second-line therapy. For premenopausal women who have undergone ovarian ablation, the use of third-line therapy with an AI becomes possible. For postmenopausal women, a wide choice of endocrine treatment options is available and an optimal sequence has yet to be determined. Options for first line therapy of metastatic disease include an AI for women who have received adjuvant tamoxifen or tamoxifen for patients who have received adjuvant anastrozole. In addition, data suggest that fulvestrant ('Faslodex'), a novel estrogen receptor (ER) antagonist that downregulates the ER protein and has no known agonist effects, is a promising therapeutic option that has shown efficacy in the treatment of postmenopausal women with advanced breast cancer. Other agents that may be used in the sequence include the steroidal AI exemestane and the progestin megestrol acetate. The widening range of adjuvant endocrine options therefore represents an opportunity to prolong patient benefits in the treatment of hormone receptor-positive breast cancer, and will require the further refinement of the optimal sequence of endocrine agents for the treatment of recurrent breast cancer. PMID- 14535532 TI - The biology of antihormone failure in breast cancer. AB - Many estrogen receptor-positive breast cancer patients initially respond to treatment with antihormonal agents such as tamoxifen, but remissions are often followed by acquisition of resistance and ultimately disease relapse. The development of a rationale for the effective treatment of tamoxifen-resistant breast cancer requires an understanding of the complex signal transduction mechanisms that contribute towards loss of antiestrogen response. Interactions between estrogen and growth factor signaling pathways have been identified in estrogen-responsive cells that are thought to reinforce their individual cellular effects on growth and gene responses. Increasing evidence indicates that abnormalities occurring in growth factor signaling pathways, notably the epidermal growth factor receptor (EGFR) signaling pathway, could dramatically influence steroid hormone action and may be critical to antihormonal-resistant breast cancer cell growth. Thus, inhibitory agents targeting growth factor receptors, or their intracellular pathway components, may prove clinically beneficial in antihormone refractory disease. One example, gefitinib ('Iressa', ZD1839), an EGFR-tyrosine kinase inhibitor, is an interesting therapeutic option that may provide benefit in the treatment of antihormonal-resistant breast cancer. Rapid progress with pharmacological and molecular therapeutic agents is now being made. Therapies that target growth factor signaling pathways may prevent development of resistance. PMID- 14535533 TI - Future directions in the endocrine therapy of breast cancer. AB - Data from the 'Arimidex', Tamoxifen, Alone or in Combination (ATAC) trial have indicated that anastrozole ('Arimidex') may supplant tamoxifen as the preferred adjuvant endocrine therapy for hormone receptor-positive, early breast cancer in postmenopausal women. The acceptability of this change in clinical practice at this time is currently under debate, and depends upon how confident we can be that 4 years' follow-up is sufficient to allow the overall risk:benefit balance to be assessed. The data supporting the benefits of sequential endocrine therapy are more certain, and if anastrozole does become the adjuvant agent of choice, the optimal sequence of endocrine agents will need to be identified for use after recurrence on anastrozole for early disease. It will be important that new endocrine options, such as the estrogen receptor antagonist fulvestrant ('Faslodex'), are suitably incorporated into these sequences. Developments such as the sequential use of endocrine therapies in the management of breast cancer have stimulated evolving concepts in our understanding of the basic cell biology of the disease. This has emphasized the importance of understanding the interactions between the different cell-signaling pathways that underlie the development of endocrine resistance. This will present a range of new opportunities for therapeutic intervention and will provide the foundation for the rational design of clinical trials. The resulting incorporation of more target-oriented approaches and newer endocrine agents into the treatment of breast cancer should stimulate the development of promising treatment paradigms and will ultimately provide further benefit to patients. PMID- 14535534 TI - Estimating conditional distributions using a method based on S-distributions reference percentile curves for body mass index in Spanish children. AB - BACKGROUND: Reference intervals are a fundamental tool for characterizing the health status in a given population and play a central role in defining diagnostic values in clinical applications. Estimation of the conditional distribution of a variable, as the body mass index (BMI), is necessary for providing reference values when there is a trend as a function of the covariate. SUBJECTS AND METHOD: We studied 1453 boys and young between 5 and 16 years old measured in a study carried out in the schools of Lleida (Spain). BMI conditional distributions with age have been derived using a new parametric method based on the one proposed by Sorribas et al. [Stat. Med. (2000) 19:697-713]. This method is based on S-distributions as a parametric model for the distribution and uses maximum likelihood estimation of the conditional distribution. RESULTS: The methods commonly used for estimating reference curves are based on a smoothing of sample quantiles using different techniques. However, these methods do not provide information on the conditional distribution of the target variable. Our method provides an estimation of such distribution and the corresponding reference curves for the quantiles as a function of a covariate, in our case age. CONCLUSIONS: The suggested methodology provides appropriate reference quantiles for the BMI. Our results allow characterizing the change in distribution within the age range considered. Besides describing a raise in BMI with age, we observe an increase in dispersion around puberty. This must be considered when using BMI as a diagnostic variable. PMID- 14535536 TI - The development of bone mass and bone strength in the mandible of the female rat. AB - The present study provides baseline data for a number of mandibular growth dimensions, specially on bone mass and bone strength, that were collected between the 21st and the 180th days of postnatal life, which are intended as a reference for researchers designing experimental studies, specially on mandibular catch-up growth, and as an aid for clinicians who must evaluate results from published animal studies for validity and potential extrapolation to the human clinical situation. Fifty weanling female Wistar (Hsd:Wi) rats were fed ad libitum a diet previously shown to allow normal, undeformed mandibular growth. Five of them were randomly selected at different times between 21 and 180 d of life. Mandibular growth was estimated directly on the right hemimandible by taking measurements between anatomical points; mandibular bone mass (calcium mass) was estimated from the mg of calcium, determined by atomic absorption spectrophotometry, present in the ashes of the left hemimandible; and mechanical properties of the right hemimandible were determined using three-point bending mechanical test. Dimensions, bone calcium mass and bone strength of the female rat mandible increased linearly from day 21 to approximately day 90. Bone growth, as expected, was more than twice when assessed from bone weight than when derived from mandibular area, length or height when the parameters were expressed as the relative increase from the mean infant condition. The growth rate of the posterior part of the mandible (behind the third molar) was almost five times greater than that of the anterior part. The rates of growth of the studied parameters showed a marked decline after day 90. ANOVA indicated that no statistical differences were found between day 90 and day 120 values. It could be concluded that the female rat mandible attains its adult size, peak bone calcium mass and bone structural mechanical properties at some point between 90 and 120 d of postnatal life. Because of the extremely high positive correlation between mandibular bone calcium mass and both mandibular area and mandibular weight, it was possible to calculate the mandibular peak bone mass from the relations 7.69 mgCa/cm2 and 0.19 mgCa/mg bone. PMID- 14535535 TI - Sexual dimorphism and postnatal growth of intrauterine growth retarded rats. AB - The intrauterine growth retardation (IUGR) is a complex phenomenon since it causes variability in postnatal growth responses. The aim of the present study was to analyze the effects of lactational rehabilitation on body and cranial growth and sexual dimorphism in IUGR rats. IUGR model was carried out by means of uterine vessels bending in pregnant rats at the first day of pregnancy. Control and sham-operated animals were also included. During suckling, IUGR and sham operated pups were cross-fostered to a control dam. The animals were x-rayed every four days, from birth to weaning. Body weight and length, and neural and facial variables were measured. The reduced placental blood flow delayed growth in pups. Facial growth was more impaired than the neural one. An incomplete catch up growth was found, since males reached sham values only in neurocranial height and females--except facial length and width--were similar to sham. Because of sex differences in growth recovery, normal sexual dimorphism in body and craniofacial pattern was not expressed in the IUGR group. This study suggests that reduced uteroplacental blood supply in the rat interferes with the full expression of normal dimorphism based on gender and confirms the impact of IUGR on postnatal growth. PMID- 14535537 TI - Changes in schoolgirls' body measurements during a 30 year period. AB - Two data-sets containing the anthropometric measurements of Estonian schoolgirls aged 7-18 were analysed, one (set A) from the late sixties, size 1627 and the other (set K) from the late nineties, size 1546. In all data sets 8 measurements- three height, three breadth measures, chest circumference and mass--were measured using the same methodology, as well as BMI was calculated. The means of all the characteristics in age groups (size 100-150 individuals) were calculated and growth curves of means were approximated using polynomial least squares approximation. The maximal velocity points were found and the growth intensities in different body build groups were compared. It was proved that the usage of cross-sectional data for estimating growth curves is suitable for estimating the velocity of acceleration in different age groups. We found that the height of Estonian girls has increased during the last 30 years by about 4-5%, the breadth measurements--by about 1-2%, chest circumference up to 8% and BMI has decreased by 3-5%. The changes are quite modest in pre-puberty, most significant for girls aged 12-16 and smaller again for girls aged 17 and more. The maximal growth velocity has shifted for all characteristics to the earlier time. Comparison of different body build groups showed that the small girls of the sixties are quite similar to small girls in the nineties, but the differences are greater in the case of large and pycnomorphic girls. PMID- 14535538 TI - Identification of canine transmissible venereal tumor cells using in situ polymerase chain reaction and the stable sequence of the long interspersed nuclear element. AB - Canine transmissible venereal tumor (CTVT) is a unique tumor that can be transplanted across the major histocompatibility complex (MHC) barrier by viable tumor cells. In dogs, CTVT grows progressively for a few months and then usually regresses spontaneously. A long interspersed nuclear element (LINE) insertion is found specifically and constantly in the 5' end of the CTVT cell c-myc gene, outside the first exon. The rearranged LINE-c-myc gene sequence has been used with polymerase chain reaction (PCR) to diagnose CTVT. However, in CTVT cells, the total length of the inserted LINE gene is not constant. In this experiment, variation in the inserted LINE gene was studied to determine which parts of the LINE sequence can be used as primers to identify CTVT cells with in situ PCR (IS PCR). The LINE gene was inserted between the TATA boxes in the promoter region of c-myc. In CTVT cells, deletions of different lengths are frequent in this gene. However, the 550-bp segment at the 5' end of the LINE-c-myc gene was stable. Thus, primers were designed to cover the stable 0.55-kb segment from the 5' end outside the first exon of the c-myc gene to the 5' end of LINE gene stable segment. With these primers and IS PCR, individual CTVT cells in formalin-fixed tissue sections and CTVT cultures were identified. Cells from other canine tumors were negative for this gene. In addition, the CTVT-specific, 0.55-kb segment was not found in any spindle-shaped cells from progressive or regressive phase CTVT. The IS PCR technique also did not detect any positive spindle-shaped cells in CTVT cell cultures. Thus, fibroblastic terminal differentiation is less likely to be a mechanism for spontaneous regression of CTVT cells. PMID- 14535539 TI - Infectious salmon anemia virus RNA in fish cell cultures and in tissue sections of atlantic salmon experimentally infected with infectious salmon anemia virus. AB - Current understanding of the etiopathogenesis of infectious salmon anemia (ISA) virus (ISAV) infection in fish comes mostly from virus detection in homogenized tissues taken from ISA-suspected mortalities. This study combined in situ hybridization (ISH) and histology to demonstrate viral RNA transcripts in different fish cell lines infected with ISAV and in tissues collected during the clinical phase of ISAV infection in Atlantic salmon. For this, a riboprobe to mRNA transcripts of ISAV RNA segment 8 was shown to detect viral mRNA in ISAV infected TO, CHSE-214, and SHK-1 cell cultures. Specific hybridization was initially detected exclusively in the nuclei of infected cells, which is consistent with the nuclear transcription of orthomyxoviruses. For use of the riboprobe on fish tissues fixed in paraformaldehyde or formalin, the conditions used to permeabilize tissues before ISH (Proteinase K or Tween 20) were first optimized. Tissues were collected 15-20 days after challenge from 7 fresh mortalities of Atlantic salmon parr (approximately 20 g) showing severe gross and microscopic lesions, consistent with ISAV infection. Reverse transcription polymerase chain reaction on tissue pools confirmed the presence of ISAV in each of the 7 fish. Of the tissues examined in each fish, the heart and liver consistently showed the strongest hybridization signal and, therefore, the most in situ virus, which was located in the endothelium of small blood vessels and in macrophage-like cells. PMID- 14535540 TI - Lead contamination of chicken eggs and tissues from a small farm flock. AB - Twenty mixed-breed adult laying hens from a small farm flock in Iowa were clinically normal but had been exposed to chips of lead-based paint in their environment. These chickens were brought to the Iowa State University Veterinary Diagnostic Laboratory, Ames, Iowa, where the concentration of lead in blood, eggs (yolk, albumen, and shell), and tissues (liver, kidney, muscle, and ovary) from 5 selected chickens was determined over a period of 9 days. Blood lead levels ranged from less than 50 to 760 ppb. Lead contamination of the yolks varied from less than 20 to 400 ppb, and shells were found to contain up to 450 ppb lead. Albumen contained no detectable amount. Lead content of the egg yolks strongly correlated with blood lead levels. Deposition of lead in the shells did not correlate well with blood lead levels. Mean tissue lead accumulation was highest in kidneys (1,360 ppb), with livers ranking second (500 ppb) and ovarian tissue third (320 ppb). Muscle contained the lowest level of lead (280 ppb). Lead contamination of egg yolks and edible chicken tissues represents a potential public health hazard, especially to children repeatedly consuming eggs from contaminated family-owned flocks. PMID- 14535541 TI - Severe persistent orf in young goats. AB - Orf (contagious ecthyma) is a viral disease of small and wild ruminants, humans, and less frequently other species. In sheep and goats, the disease is characterized by the formation of vesiculo-proliferative lesions in the skin of lips and nostril. Here, a form of generalized orf in 16 goat kids from 2 different locations in west Texas is described. The disease was characterized by multifocal, severe, proliferative dermatitis that persisted from about 2 months of age until the goat kids were euthanized 3 months later. All affected goats were Boer or Boer crosses under 1 year of age. The mean immunoglobulin concentration in sera of affected goats was elevated compared with healthy control goats. Severe to moderate lymphadenomegaly of the nodes draining the areas of the skin affected with orf lesions was present in all 16 goat kids. Suppurative arthritis, chronic fibrinous pneumonia, and premature thymic involution were found in 3, 5, and 7 of the goat kids, respectively. The skin lesions of 3 goat kids were infested with larvae of the opportunistic black garbage fly (Ophira sp.). The orf virus was identified in skin lesions by isolation in Marbin-Darby ovine kidney cells, electron microscopy, and amplification of viral DNA by polymerase chain reaction. The orf virus was not detected in peripheral blood or lymph node mononuclear cells of any of the goats. Cross-neutralization experiments showed that an ovine orf virus antiserum raised in sheep was more effective in neutralizing a sheep orf virus isolate than a caprine orf virus isolate. The clinical and epidemiological characteristics of these orf cases may be the result of susceptibility factors within some individuals of the Boer breed of goats. PMID- 14535542 TI - Ascaridoid nematodes and associated lesions in stomachs of subadult male northern fur seals (Callorhinus ursinus) on St. Paul Island, Alaska: (1987-1999). AB - Stomachs from subadult male northern fur seals (SAMs) (Callorhinus ursinus) (n = 5,950) from St. Paul Island, Alaska, were examined from 1987 to 1999 for lesions and parasites. On gross examination of these stomachs, parasite nodules were evident in 92% of the stomachs and active and healing ulcers were found in 14% and 10%, respectively. Prevalence of nematodes from 1987 to 1997 was 88% for 5,700 SAMs for which numbers of parasites were estimated but not identified to the genus level. All nematodes recovered from 250 SAMs examined in 1998 (n = 124; 99% infected) and in 1999 (n = 126; 91% infected) were identified and counted. Prevalences in 1998 and 1999 were 5% and 0% for Anisakis spp., 52% and 18% for Contracaecum spp., and 96% and 89% for Pseudoterranova spp., respectively. PMID- 14535543 TI - Preparation and characterization of polyclonal and monoclonal antibodies against Lawsonia intracellularis. AB - Proliferative enteropathy is an intestinal infectious disease caused by the obligate intracellular bacterium Lawsonia intracellularis. Immunohistochemistry staining has superior sensitivity over hematoxylin and eosin and silver staining for detecting L. intracellularis in histological sections. A L. intracellularis specific monoclonal antibody (MAb) produced in the UK (IG4 MAb) has been described in the literature. However, no monoclonal or polyclonal antibodies are commercially available. Therefore, the objective of this study was to produce and characterize new polyclonal and monoclonal antibodies against L. intracellularis that are suitable for diagnostic use. The new monoclonal (2001 MAb) and polyclonal antibodies (1999 PAb) were compared with the IG4 MAb using Western blot analysis of outer membrane proteins (OMPs) of 6 L. intracellularis isolates, Bilophila wadsworthia and Brachyspira hyodysenteriae and using immunohistochemistry of known positive and negative histologic samples and pure cultures of L. intracellularis, B. wadsworthia, B. hyodysenteriae, Salmonella choleraesuis, S. typhimurium, and Escherichia coli K88. Immunogold staining using 2001 MAb was performed to show the specificity of the antibody against an L. intracellularis surface protein. Western blot analysis showed that the 2001 MAb targeted an OMP of 77 kD, which made it different from the IG4 MAb that targeted an 18-kD OMP. The immunogold stain demonstrated the specificity of the 2001 MAb to a surface protein of L. intracellularis. The polyclonal antibody (1999 PAb) targeted 5 OMPs (77, 69, 54, 42, and 36 kD). Both the 2001 MAb and 1999 PAb stained known positive, but not negative, histologic samples. Both the 2001 MAb and 1999 PAb reacted with a pure culture of L. intracellularis but not with any other common enteric pathogens. These two new antibodies will be useful for immunodiagnosis of L. intracellularis. PMID- 14535544 TI - Use of a Japanese quail fibrosarcoma cell line (QT-35) in serologic assays to determine the antigenic relationship of avian metapneumoviruses. AB - The ability of a Japanese quail fibrosarcoma cell line (QT-35) to support the replication of avian metapneumoviruses belonging to the 3 subgroups A (14/1 virus), B (Colorado virus), and C (Hungary virus) enabled the development of assays for the detection and evaluation of virus-specific antibodies. On the basis of the results of enzyme-linked immunosorbent assay (ELISA), plaque reduction neutralization assay (PRNA), immunofluorescent assay (IFA), and Western blot analysis, some degree of antigenic cross-reactivity was observed between prototype viruses belonging to each of the 3 subgroups A, B, and C. The antigen produced in QT-35 cells was found to be superior with respect to its reactivity with virus-specific antibodies, as determined when used in ELISA and IFA. Standardization of both the input virus and the virus-specific antibodies in PRNA enabled a more detailed analysis of the antigenic relationship between these viruses. Specifically, it was observed that 14/1 virus shared more neutralizing regions with Hungary and Colorado viruses than did either of these viruses with 14/1 virus. In addition, Hungary virus shared comparatively fewer neutralizing epitopes with the Colorado virus than did 14/1 virus. Western blot analysis of the reactivity patterns of virus antigen, produced in QT-35 cells, with subgroup specific antibodies identified a cross-reactive protein migrating at approximately 18 kD. These assays and the information from the Western blot will enable further analysis of avian metapneumovirus isolates to determine antigenic relationships. PMID- 14535545 TI - Multiple cutaneous leiomyomas in the perineum of a horse. AB - Multiple cutaneous masses developed in the perineum of a 14-year-old Saddlebred stallion over a period of approximately 5 years. Clinically, the masses ranged in size from 3- to 9-mm diameter and were not ulcerated, painful, or pruritic. Three of the masses were surgically excised and submitted for microscopic evaluation. The masses were dome shaped to nodular, located in the superficial dermis, and composed of haphazardly arranged bundles of plump spindle-shaped cells. The tumor cells immunoreacted with monoclonal antibodies directed against desmin, muscle specific actin, and smooth muscle actin, confirming a smooth muscle origin. Multiple cutaneous leiomyomas have not been reported previously in horses. PMID- 14535546 TI - Neuronal ceroid-lipofuscinosis in a Labrador Retriever. AB - An 8-year-old Labrador Retriever with an 11-month history of progressive partial seizures and necropsy examination findings characteristic of the lamellar form of canine neuronal ceroid-lipofuscinosis (NCL) is presented. The clinical, light microscopic, and ultrastructural features of this case most closely resemble human adult-onset NCL (Kufs disease). This is the first report of NCL occurring in the Labrador Retriever breed. PMID- 14535547 TI - Comparison of the diagnostic sensitivity of a commercially available culture kit and a diagnostic culture test using Diamond's media for diagnosing Tritrichomonas foetus in bulls. AB - A number of different culture media have been described for use in the diagnosis of Tritrichomonas foetus infection in bulls, and recently, a commercial culture kit has become available. The objective of this study was to compare the sensitivity of 2 culture-based diagnostic tests for T. foetus in bulls. One test used a commercial kit for transport and culture of the samples. The other test used a thioglycollate transport medium (TFTM) for transport and a modified Diamond's medium (MDM) for culture of the samples. Twenty-one bulls infected with T. foetus were sampled repeatedly. On each sampling day, samples collected from the left and right sides of the bull were tested with one of the 2 diagnostic tests being compared. The effect of the type of diagnostic test on the outcome of the test was evaluated with a chi-square test for the calculated odds ratio. Because repeated tests from the same bull cannot be considered independent measures, unadjusted chi-square tests were adjusted for the effect of clustering by bull. Samples tested using the commercial kit were 6.95 times as likely to be positive as samples tested with a diagnostic test using MDM (P < 0.001). PMID- 14535548 TI - 16S ribosomal RNA sequence-based identification of veterinary clinical bacteria. AB - This study evaluated 16S rRNA gene sequence analysis methods as tools for identification of 22 phenotypically difficult to identify veterinary clinical bacterial isolates in a veterinary diagnostic laboratory. The study compared 16S rRNA gene sequencing and conventional phenotypic identification methods. Using 16S rRNA full-gene sequencing, 95% (21/22) of the isolates were identified to the genus level and 86% (19/22) to the species level. The conventional or commercially available manual identification phenotypic characterization methods presumptively identified 91% (20/22) of the isolates to the genus level and 1 isolate to the species level. However, only 55% (12/22) or 4.5% (1/22) of the phenotypic identifications were correct at the genus or species level when they were compared with the 16S rRNA full-gene sequencing. This study also compared 16S rRNA full-gene and partial-gene sequencing. The results demonstrated that the best 16S rRNA gene-sequencing approach is full-gene sequencing because it gives the most precise species identification. Sequencing of the variable regions 1, 2, and 3 of the 16S rRNA gene could be used for tentative identification because the ability of this sequencing to identify bacteria to the genus level is similar to that of the 16S rRNA full-gene sequencing. This method identified only 14% (3/22) isolates differently to the species level compared with the 16S rRNA full gene sequence. Sequencing of the variable regions 7, 8, and 9 is not recommended because it gives more ambiguous identifications. The cost of a 16S RNA full-gene sequencing analysis was Can 160 dollars and Can 60 dollars for a partial 16S rRNA gene sequence, i.e., sequencing of variable regions 1, 2, and 3 or variable regions 7, 8 and 9. PMID- 14535549 TI - Rhodococcus equi and genetic susceptibility: assessing transferrin genotypes from paraffin-embedded tissues. AB - Rhodococcus equi is a bacterial pathogen, ubiquitous in the soil, that infects many foals and is lethal to some. Transferrin is an iron-binding protein that has bacteriostatic properties in the blood. Transferrin is also highly polymorphic in most species, with 15 variants identified for horses using biochemical methods, and may be responsible for variation in susceptibility to bacterial pathogens. The objectives of this study were 1) to compare biochemical typing with DNA typing of transferrin, 2) to determine if transferrin DNA from archival paraffin embedded tissue samples can be recovered and typed with molecular methods, and 3) to determine if there was an association between foal death caused by R. equi and transferrin type. Comparing biochemical methods and DNA sequencing for 41 horses demonstrated correspondence between the typing methods. The allele frequency of archival paraffin-embedded tissue samples from 34 Thoroughbred foals that succumbed to R. equi showed an excess of the F allele and a deficiency of the D allele (P < 0.05). Year of collection (P > 0.8), age of foal (P > 0.3), and sex of foal (P > 0.6) were not statistically associated with transferrin type. The archival material was successfully transferrin typed using DNA sequencing, and there may be an association between foal death caused by R. equi and transferrin type. PMID- 14535550 TI - Valvular endocarditis associated with Helcococcus ovis infection in a bovine. AB - A 12-month-old Angus bull calf with a history of fever and lethargy of several weeks' duration was necropsied. Macroscopic findings included general dehydration, congestion, and edema within the craniodorsal lobes of the lung, multifocal ecchymotic hemorrhages on the dorsal epaxial and gluteal muscles, bloody ingesta within the gastrointestinal tract, and a 4- x 4- x 5-cm irregular plaque located on the right atrioventricular heart valve. Microscopically, there were focally extensive pulmonary alveolar infiltrates of histiocytes and neutrophils, large numbers of necrotic hypereosinophilic hepatocytes located within the centrilobular and midzonal regions of the liver, and, within the plaque from the right atrioventricular valve, a large mass formed by abundant laminated fibrin that contained numerous small multifocal aggregates of gram positive cocci. This report describes the first apparent isolation of Helcococcus ovis from cattle. PMID- 14535551 TI - The seroprevalence of Johne's disease in Georgia beef and dairy cull cattle. AB - Beef and dairy cattle serum samples, collected during 2000 at sale barns throughout Georgia, were obtained from the Georgia State Brucellosis Laboratory and were used to conduct a retrospective epidemiological study. Statistical samplings of 5,307 sera, from over 200,000 sera, were tested for antibodies to Mycobacterium avium ssp. paratuberculosis, (Johne's disease) using a commercial enzyme-linked immunosorbent assay test kit. An overall period seroprevalence in all classes of cattle tested was 4.73%. The period seroprevalence in dairy cattle was 9.58%, in beef cattle it was 3.95%, and in cattle of unknown breed it was 4.72%. It was concluded that the seroprevalence of Johne's disease in cull beef and dairy cattle in Georgia is economically significant. PMID- 14535552 TI - Cyanide toxicosis in goats after ingestion of California Holly (Heteromeles arbutifolia). AB - Cyanogenic glycosides are found in many native and naturalized plants throughout North America. The glycosides themselves are not toxic, but they yield hydrogen cyanide (hydrocyanic or prussic acid) when they are hydrolyzed by beta glycosidases, either as a result of injury to the plant cells or by microbial action in the rumen. Hydrogen cyanide is readily absorbed from the gastrointestinal tract. Cyanide ion binds with iron in cytochrome oxidase, interfering with cellular respiration. The clinical effects are peracute, often resulting in death less than 1 hour after ingestion. This study describes a case that resulted in significant morbidity and mortality in a herd of goats after exposure to California holly (Heteromeles arbutifolia). PMID- 14535553 TI - Simple capillary electrophoretic determination of soluble oxalate and nitrate in forage grasses. AB - A simple capillary electrophoretic method is described for the simultaneous determination of soluble oxalate and nitrate in forage grasses. Grass samples were ground and extracted with water. The extracts were filtered and submitted to capillary electrophoresis. Electrophoresis was performed in a 75 microm x 50 cm fused silica capillary with 30 mM sodium sulfate containing an electroosmotic flow modifier under constant voltage at -8 kV. Separated oxalate and nitrate were detected with direct UV absorption at 214 nm. The present method can be used for routine monitoring of the concentration of soluble oxalate and nitrate in grasses. PMID- 14535554 TI - Cerebral cysticercosis by Taenia crassiceps in a domestic cat. AB - Cerebral cysticercosis by Taenia crassiceps was diagnosed in an adult female domestic shorthair cat. The animal was euthanized 6 weeks after the initial presentation with signs of vomiting, lethargy, and ataxia. The disease took an intermittent relapsing course with the neurological signs progressing eventually to recumbancy and coma. At necropsy, numerous cysticerci were found in the dilated left lateral ventricle and the adjacent brain parenchyma. The cysticerci were identified as metacestodes of T. crassiceps larvae based on size and morphology of the cysts; shape, number, and size of the rostellar hooks; and mode of proliferation, including endogenous and exogenous budding. Cerebral cysticercosis by T. crassiceps is rare in atypical intermediate hosts and has not been described in cats. PMID- 14535555 TI - Nested multiplex RT-PCR for detection and differentiation of West Nile virus and eastern equine encephalomyelitis virus in brain tissues. AB - A traditional nested reverse transcription-polymerase chain reaction (RT-PCR) assay specific for eastern equine encephalomyelitis (EEE) virus was designed to multiplex with a previously described West Nile (WN) virus nested RT-PCR assay. Differentiation of EEE and WN was based on base pair size of the amplified product. One hundred fifty-seven mammalian and avian brain tissues were tested by EEE/WN nested multiplex RT-PCR, EEE nested RT-PCR, and WN nested RT-PCR, and results were compared with other diagnostic test results from the same animals. Serological and virus isolation testing confirmed the results of the multiplex PCR assay. When compared with cell culture virus isolation, the multiplex assay was shown to be more sensitive in detecting the presence of EEE or WN virus in brain tissues. The multiplex assay was shown to be sensitive and specific for North American EEE and WN and provided a rapid means of identifying both viruses in brain tissues. No apparent sacrifice in sensitivity was observed in the multiplex procedure compared with the individual EEE and WN nested RT-PCR assays. Data collected from an additional 485 multiplex RT-PCR tests conducted during the summer and fall of 2002 further support the validity of the procedure. PMID- 14535556 TI - Efficacy of resorbable plates for reduction and stabilization of laryngeal fractures. AB - We evaluated the efficacy of resorbable reconstruction plates (polylactic acid copolymer) for the open reduction and stabilization of displaced laryngeal fractures. Both MacroPore and Leibinger reconstruction plates were used with equal ease of application in 3 adult male patients. We found the plating system to be especially effective for the reduction of comminuted cricoid fractures. Adequate skeletal stabilization allowed early resumption of phonatory and respiratory function without long-term intraluminal stenting for skeletal support. No complications of hematoma, seroma, or infection were experienced. Resorbable plates appear to be relatively safe and useful for internal fixation of both cartilaginous and ossified parts of the larynx, allowing rapid rehabilitation and return of function. PMID- 14535557 TI - 585-nm pulsed dye laser treatment of glottal dysplasia. AB - Management of glottal dysplasia can be difficult and often results in a suboptimal treatment outcome. The surgeon and patient must cooperatively balance decisions regarding the effects of possible malignancy, vocal dysfunction, and recurrences leading to multiple use of general anesthetics. A pilot study was done in 57 cases (36 patients and 97 vocal folds) without complication to evaluate the effectiveness of a 585-nm pulsed dye laser (PDL; 450-micros pulse width, 19 to 76-J/cm2 fluence, 1- to 2-mm spot size) in the treatment of vocal fold keratosis. Forty of the 57 cases had bilateral treatment. Phonomicrosurgical resection was done in 35 of the 57 cases after PDL treatment. Of this group, 10 cases were found to have hyperplasia, 21 dysplasia, 4 carcinoma in situ, and 1 carcinoma. One patient had phonomicrosurgical resection before PDL treatment. In 21 of the 57 cases, the disease was irradiated without resection (4 unilateral lesions and 17 bilateral lesions). Approximately 80% of the patients in this series had a greater than 70% reduction in the size of the lesion with the use of the PDL irrespective of whether they underwent resection. Clinical observation revealed no new anterior commissure web formation despite bilateral anterior commissure treatment in 28 of the 57 cases. The PDL enhanced the epithelial excision by improving hemostasis and by creating an optimal dissection plane between the basement membrane and the underlying superficial lamina propria. In this initial trial, the PDL provided relatively safe and effective treatment for glottal dysplasia. Analysis of patterns of recurrence will require longer follow up. PMID- 14535558 TI - Preoperative and postoperative voice in Tis-T1 glottic cancer treated by endoscopic cordectomy: an additional issue for patient counseling. AB - Radiotherapy contends with endoscopic surgery for the role of treatment of choice for Tis-T1 glottic cancer. The amount of vocal cord to be surgically removed logically depends on the surface and deep extension of the neoplasm. Thus, a prerequisite for proper management includes an analysis of the voice changes after each of the progressive types of cordectomy described in the European Laryngological Society Classification. Between January 1998 and December 2000, 89 patients with glottic cancer (8 Tis, 63 T1a, 18 T1b) underwent different types of endoscopic cordectomy. Perceptual analysis (GRBAS scale); objective analyses of jitter, shimmer, and noise-to-harmonics ratio; and subjective (Voice Handicap Index) evaluation of voice were performed in 51 patients. Statistical evaluation of preoperative and postoperative objective results by analysis of covariance, as well as perceptual and subjective data, showed significant voice improvement after type I and II cordectomies, with the voice attaining nearly normal parameters. By contrast, after type III, IV, and V cordectomies, the vocal outcome was not significantly different from the preoperative pattern. It can therefore be concluded that type I and II resections, whenever indicated, are adequate procedures even for professional voice users. By contrast, accurate counseling is mandatory before type III, IV, and V cordectomies. PMID- 14535559 TI - Congenital bilateral adductor vocal cord paralysis. AB - Bilateral adductor vocal cord paralysis (BAdP), presenting with features of laryngeal incompetence, is a rare form of congenital bilateral vocal cord paralysis, and only 2 small series of BAdP have previously been published. Three cases are reported here. The BAdP occurred as an isolated abnormality in 1 child, and was associated with a recognizable syndrome (Robinow's syndrome and 22q deletion) in the other 2 children. Gastrostomy tube feeding was required in 2 children, who both remain gastrostomy tube-dependent at 26 months and 10 years 9 months of age. The child with Robinow's syndrome received parenteral nutrition until 2 months, but was then able to feed orally after partial improvement in vocal cord function. The global impairment in vocal cord constrictor function observed in these 3 children is consistent with the site of lesion's being at the level of the laryngeal constrictor motoneurons in BAdP. PMID- 14535560 TI - Morphological comparison between neonatal and adult human tongues. AB - There are currently no descriptions of neonatal tongue anatomy. Therefore, there have been no reports on the morphological differences between it and the adult tongue that would suggest its suitability for suckling. Serial coronal sections of a neonatal tongue were used to create a 3-dimensional model that was compared to that of the adult tongue. Compared to the adult human tongue, the neonatal tongue was found to contain 1) considerably less fat and soft tissue; 2) a thinner mucosa; 3) relatively enlarged extrinsic musculature; 4) a less-developed superior longitudinal muscle, resulting in a flat dorsal surface; and 5) attachments between the extrinsic muscles and the transverse muscle group that have not been identified in the adult tongue. The particular structure of the neonatal tongue suggests how the neonatal tongue is specialized for suckling. PMID- 14535561 TI - Two-stage removal of an impacted foreign body with an epoxied anchor. AB - A stone impacted in a child's external auditory canal had defied all conventional means of removal. It was extracted successfully after attachment of a specially formed metal anchor with epoxy glue. The technique of and rationale for this approach are discussed. PMID- 14535562 TI - Zoledronic acid inhibits osteoclastogenesis in vitro and in a mouse model of inflammatory osteolysis. AB - This study assessed effects of the bisphosphonate zoledronic acid (ZLNA) on osteoclastogenesis. To assess the effect of ZLNA on osteoclast formation in vitro, we cultured mouse bone marrow cells under conditions that promote osteoclastogenesis. Administered at concentrations from 10(-6) to 10(-9) mol/L, ZLNA led to a dose-dependent inhibition of osteoclastogenesis. Combined TUNEL staining and histochemical staining for tartrate-resistant acid phosphatase showed that ZLNA induced apoptosis in osteoclasts and monocytic precursor cells. To study the effects of ZLNA in vivo, we placed keratin particles onto the surface of the parietal bone of mice to induce localized inflammatory bone resorption. Three experimental groups received daily subcutaneous injections of ZLNA (1, 3, or 10 microg/kg body weight) from 4 days before surgery until 5 days after keratin implantation. The ZLNA significantly reduced osteoclast recruitment in a dose-dependent manner, but did not affect the degree of inflammation or the mineral apposition rate. PMID- 14535563 TI - Systematic direct laryngoscopy: the Lindholm laryngoscopes. AB - The authors, each with 40 years of experience in laryngology, aim to lay out the general principles and details of a systematic method of direct laryngoscopy for adults, children, and infants. Advances in laryngoscope design and application, advantages of telescopes, use of the operating microscope, and principles of modem anesthesia are highlighted. Particular reference is made to classification of laryngoscopes, advantages of Lindholm laryngoscopes, suspension laryngoscopy, the principles of biopsy, and problems of laryngoscopy. The difficult airway and the obstructed airway are discussed in detail. With the recent renewed interest in investigation and treatment of laryngeal problems and a better understanding of laryngeal physiology and voice production, the future will, no doubt, see new procedures to treat and restore laryngeal function. The fundamentals in this report form a basis for direct laryngoscopy, endolaryngeal microsurgery, laser surgery, and phonosurgery. PMID- 14535564 TI - Successful treatment of idiopathic laryngotracheal stenosis by resection and primary anastomosis. AB - We studied the early and long-term response of idiopathic laryngotracheal stenosis (ILTS) to treatment by 1-stage laryngotracheal or tracheal resection and reconstruction in 73 patients. Nineteen of the 72 (26%) noted no difference in their voices or any difficulty in breathing after reconstruction. Forty-seven (64%) described loss of ability to project their voices as loudly as before or noted some difficulty in singing as well as they did before. Five (7%) had various degrees of dyspnea or stridor on effort. One needed continued dilation. The median follow-up was 8 years. Surgical treatment gave good or excellent results in 90% of these patients with ILTS. Recurrence or progression of stenosis was not evident. PMID- 14535565 TI - Aeration following intact canal wall mastoidectomy. AB - This study examines the prevalence and extent of re-aeration of the mastoid cavity following intact canal wall (ICW) mastoidectomy. Temporal bone computed tomography scans from patients with prior unilateral ICW mastoidectomy were identified. Three-dimensional volume reconstruction of the temporal bone was performed to measure aeration bilaterally. Thirty-five scans were analyzed; 16 (46%) showed good aeration in the operated ear and 19 showed poor aeration. The aeration (volume) in the surgical ears and the contralateral ears was significantly less than that in subjects without a history of ear disease. Those with poor aeration were more likely to require additional surgery. For temporal bone pairs with greater volume in the operated ear, the average difference was 1.3 cm3. Surgical creation of a mastoid cavity does not produce a large increase in aeration as compared with the contralateral ear. Following surgery, mastoid opacification may presage recurrent disease. Routine use of mastoidectomy in an attempt to improve aeration is not advocated. PMID- 14535566 TI - Tubes of vascularized cartilage used for replacement of rabbit cervical trachea. AB - This study was performed to evaluate the healing process of vascularized cartilage tubes after reconstruction of circumferential tracheal defects in rabbits. Vascularized cartilage was obtained by applying ear cartilage to a vascularized fascia flap in the lateral thoracic area in 10 animals. Five animals (control group) were used to evaluate the viability of the vascularized cartilage after 2 weeks. Circumferential tracheal defects were reconstructed in 5 animals by means of tubes of vascularized cartilage with preservation of the established blood supply around the grafts. The experimental animals were followed until signs of dyspnea became apparent. After follow-up and euthanasia, cartilage viability and regeneration of respiratory epithelium were evaluated. A viable plate of cartilage that was intensely attached to the vascularized fascia flap was found in the control animals. The animals from the experimental group showed dyspnea after a mean follow-up period of 22.6 days because of cartilage necrosis with loss of airway support. Cartilage graft revascularization and remucosalization were limited to 18.1% of the initial surface area of the cartilage tube. Mucosal coverage was seen at the anastomoses, whereas the middle part of the cartilage tube underwent necrosis. We conclude that tubes of autologous cartilage show problematic healing when placed inside the airway. Migration of vascularized connective tissue, migration of respiratory epithelium, and preservation of the viability of the cartilaginous graft were limited to a short segment at the anastomotic sites. PMID- 14535567 TI - Pediatric intraparotid Castleman's disease. AB - Castleman's disease (CD) is a rare, benign lymphoepithelial disease of unknown cause that presents most commonly in the mediastinum. There are 2 histologic types of CD: the hyaline vascular type and the plasma cell type. In the head and neck, 98% of these lesions are of the hyaline vascular type. The differential diagnosis of CD includes reactive lymphadenopathy, lymphoproliferative disorders, lymph node metastasis, and other conditions. Approximately 80 cases of CD have been reported in children; head and neck involvement in children is extremely rare. A case of a child with intraparotid CD is presented; the clinical course, histopathologic presentation, radiographic findings, and management of CD are reviewed. PMID- 14535568 TI - Worldwide distribution of Waardenburg syndrome. AB - To clarify the multiracial occurrence of Waardenburg syndrome, we present a case series and literature review. A computerized review of the English-language literature was conducted to assess the distribution of reported occurrences of Waardenburg syndrome in populations around the world. We detail the clinical features of 2 family cohorts: one of Western European origin and the other from South Asia. A computerized literature review found sporadic cases of the syndrome in many ethnic groups, including Japanese, Taiwanese, and Middle Eastern families. The highest reported incidence is among Kenyan Africans. Waardenburg syndrome accounts for between 2% and 5% of cases of congenital deafness. It was first described in Northern European cohorts and is widely identified in fair skinned populations. We hope to raise awareness of the worldwide distribution of this important cause of hearing loss. PMID- 14535569 TI - Apnea spells in an infant with vallecular cyst. PMID- 14535570 TI - Hearing impairment in Usher syndrome type II. PMID- 14535571 TI - Why is chemical brain injury ignored? Pondering causes and risks. PMID- 14535572 TI - Effect of air pollution on respiratory health in Indonesia and its economic cost. AB - A total of 16,663 pairs of junior high school students and their mothers in Indonesian cities were surveyed, using a self-administered questionnaire, to measure the effect of air pollution on respiratory health and the cost of associated illness. Multiple regression analysis showed that the prevalence rates of the symptoms of cough, phlegm, persistent cough, wheezing without a cold, and asthma, in the student and mother groups, were significantly correlated with the nitrogen dioxide (NO2) emitted along large roads near their residences, and to a lesser extent with smoking. In Central Jakarta and Tangerang, where the average NO2 concentrations were highest (37 ppb and 31 ppb, respectively), reduction of NO2 to a proposed level of 25 ppb could yield savings in mean direct out-of pocket expense per capita for treatment of the above symptom(s) of 15,639-18,165 Indonesian rupiah (6.80-7.90 US dollars), and reduce average work/school days lost per capita by 3.1-5.5 days. PMID- 14535573 TI - Environmental exposure to asbestos in eastern Turkey. AB - In this study, the authors investigated the prevalence of asbestos-related disorders among the inhabitants of Guzelyurt, a town in Malatya, located in eastern Turkey. The authors examined river bed, white soil, and stucco samples taken from various locales in Guzelyurt, and they confirmed the presence of tremolite and chrysotile asbestos fibers. Subjects (N = 920; 449 males and 471 females) were examined by photofluoroscopy. Eighty-five patients (9.2%) had asbestos-related radiological findings; risk increased with age. Calcified pleural plaques were seen more frequently in individuals > or = 50 yr of age, compared with younger subjects (p < 0.01). Asbestos-related disorders were prevalent in the inhabitants of Guzelyurt, the population of which is exposed environmentally to asbestos--primarily the result of the stuccoing and whitewashing of houses with soil that contains asbestos. PMID- 14535574 TI - Relationship of jute dust to interstitial fibrosis in rat lung. AB - The relationship between jute dust and lung interstitial fibrosis was studied by instilling groups of rats, via trachea, with jute dust and comparing the results with those for positive (quartz) and negative (saline) controls. The rats were sacrificed at regular intervals and their lungs and hilar lymph nodes were analyzed for collagen content and morphologic changes. The earliest changes consisted of alveolar edema, increased numbers of intraalveolar macrophages, and marked thickening of the interalveolar septa, with mixed cellular infiltrates. Moderate thickening of the alveolar walls and the zones around the peribronchioles was seen in the test groups at 6 mo. After 12 mo, some fibrosis of the alveoli walls and peribronchiole zones occurred. Interstitial cellular nodules were observed occasionally, composed mainly of dust particles, fibroblasts, reticular fibers, and collagen fibers. The collagen content in the lungs of the jute dust groups was significantly higher than for the saline control group for all test periods. The authors conclude that jute dust may induce lung interstitial fibrosis. PMID- 14535575 TI - Assessment of respiratory symptoms and asthma prevalence in a U.S.-Mexico border region. AB - The authors studied children who were 10-12 yr of age and who resided in sister cities in a U.S.-Mexico border region to determine the prevalence of asthma and respiratory symptoms. The relationship of symptoms to ambient levels of particulate matter less than 10 microm in diameter (PM10), and to several indoor environmental conditions, was assessed. The study was conducted in the border cities of Ambos Nogales (Nogales, Arizona [United States], and Nogales, Sonora [Mexico]). At the beginning of the 11-wk study, during the autumn of 1996, 631 students and their parents completed baseline questionnaires. While in school, the children completed daily symptom diaries and daily peak expiratory flow maneuvers. PM10 values and daily temperatures were also measured. The authors found that the prevalence of self-reported asthma among 5th-grade students was comparable on both sides of the border (i.e., 7.6% on the Arizona side and 6.9% on the Sonora side). Wheezing was a frequent complaint (29.5-35.6%), as was cough (16.8-29.6%). Smoking in the home was common on both sides of the border, and it was associated with a greater occurrence of self-reported asthma and respiratory complaints. Increased respiratory symptoms were also associated with increased ambient PM10 levels. The prevalence of respiratory symptoms such as wheezing and frequent cough among all children in this study, combined with the limitations inherent in self-reporting, suggest that asthma may actually be more prevalent than has been previously reported. PMID- 14535576 TI - Ecologic study of water hardness and cerebrovascular mortality in Japan. AB - Elevated levels of water hardness have been associated with reduced mortality from cardiovascular and cerebrovascular diseases. In this ecologic study, the authors examined the relationship between water hardness and cerebrovascular mortality in the 44 municipalities of Osaka Prefecture, Japan. Data on cerebrovascular death rates and water hardness in each municipality were obtained from public records; information on socioeconomic and health-care status in each municipality was incorporated with multiple logistic-regression analysis. The authors were unable to substantiate an inverse association of water hardness with cerebrovascular mortality--although the adjusted odds ratio in which the highest vs. lowest tertiles of water hardness were compared was 0.90 (95% confidence interval = 0.79, 1.02) in females. In addition, the difference between the sexes was not significant. In summary, the authors found no evidence that water hardness protected against cerebrovascular mortality. PMID- 14535577 TI - Creatine phosphate kinase elevations signaling muscle damage following exposures to anticholinesterases: 2 sentinel patients. AB - In this study, the authors describe 2 patients who experienced confirmed exposures to anticholinesterases that commenced in the 1970s. Subsequently, elevations in creatine phosphate kinase (CPK) were initially detected more than a decade following the first acute exposure. Beginning in the early 1980s, the patients suffered from progressive generalized muscle weakness, chronic fatigue, myopathy, neuropathy, and severe neurobehavioral impairments. Previous occupational exposures included pyridostigmine, as well as isopropyl methylphosphonofluoridate (percutaneous lethal dose [LD50] < 28 mg/kg body weight), and 1 patient had exposure to agricultural organophosphates. The authors hypothesize that the workers' CPK elevations, first detected more than a decade following acute exposures to anticholinesterases, were sentinel events for impending muscle damage and necrosis. Many Gulf War veterans with Gulf War disease who reported exposures to anticholinesterases 1 decade earlier currently suffer from vague neuromuscular and cognitive impairments. Therefore, medical programs for Gulf War veterans with Gulf War Syndrome should include surveillance for elevated CPK, abnormalities of neuromuscular conduction, and genetic susceptibility, and they should promote therapeutic trials for palliation. PMID- 14535578 TI - Blood lead levels and risk factors for lead poisoning among children in a Mexican smelting community. AB - The authors evaluated mean blood lead levels (BLLs) and the prevalence of elevated BLLs in children 1-6 yr of age living in Torreon, Mexico, and assessed risk factors for lead exposure in these children. The study involved a simple random sample of households in the area around a local smelter, as well as a 2 stage cluster sample of neighborhoods and households in the remainder of Torreon. The geometric mean BLL of children in this study (N = 367) was 6.0 microg/dl (95% confidence interval [CI] = 5.2, 6.8) (0.29 microM/l [95% CI = 0.25, 0.33]). Twenty percent of the children had BLLs > or = 10 microg/dl (0.48 microM/l), and 5% had BLLs > or = 20 microg/dl (0.97 microM/l). In multivariate analyses, distance from the smelter, amount of income, and education level of the primary caregiver predicted BLLs. In the environmental risk factor subsample (n = 124), dust and soil lead levels were associated with BLLs and distance from the smelter. BLLs in this study were moderately high, but the levels were lower than those in other smelting communities prior to remediation. PMID- 14535579 TI - Children's blood lead levels after the phasing out of leaded gasoline in Shantou, China. AB - Shantou, a city in the People's Republic of China, has a central urban population of approximately 700,000 and a large number of registered vehicles (motorcycles = 256,600; other motor vehicles = 261,000). Shantou is designated as 1 of the 5 Special Economic Zones in China. The use of leaded gasoline was prohibited in the city at the end of 1998. Blood samples for lead analysis were collected from children aged 1-5 yr during the summers of 1999-2001, and the samples were analyzed using graphite furnace atomic absorption spectrometry. In 1999, 44.3% (147/332) of the children had mean blood lead levels in excess of 100 microg/l. The percentage of children with blood lead levels that exceeded 100 microg/l was reduced to 35.8% (227/635) and 23.0% (105/457) in 2000 and 2001, respectively. Average blood lead levels declined from 104 microg/l, to 94 microg/l, and 79 microg/l during the 3-yr period, respectively; the respective concordant geometric means were 97 microg/l, 85 microg/l, and 71 microg/l. Mean blood lead levels for the children studied decreased significantly (p < 0.05) every year. Among all children tested during the 3-yr period, there were no significant differences by gender and no significant differences between children who were between the ages of 2 and 3 yr; however, those who were 4 yr of age had blood lead levels that were significantly (p < 0.05) higher than levels in those 2 and 3 yr of age. PMID- 14535580 TI - Canine transmissible venereal tumour: cytogenetic origin, immunophenotype, and immunobiology. A review. AB - Canine transmissible venereal tumour (CTVT) is the only known naturally occurring tumour that can be transplanted as an allograft across major histocompatibility (MHC) barriers within the same species, and even to other members of the canine family, such as foxes, coyotes and wolves. The progression of this tumour is unique in that, it follows a predictable growth pattern. In natural and experimental cases, the growth pattern includes progressive growth phase, static phase and regression phase, and this is followed by transplantation immunity in immunocompetent adults, while metastasis occurs in puppies and immunosuppressed dogs. Because of the uniqueness of CTVT transmission and progression, experimental investigations of various aspects of the biology of CTVT have been used to provide clues to the immunobiology of both animal and human tumours. This review examines the current state of knowledge of the aspects of the cytogenetic origin, immunophenotype, immunobiology and immunotherapy of CTVT. PMID- 14535581 TI - First cases of animal diseases published since 2000. 1. Dogs. AB - In this first article of a series of papers listing first case reports of animal diseases published since 2000, the following 19 cases of dog diseases are discussed: Blastomycotic granuloma involving the cranial vena cava. Congenital myocardial hamartoma. Discospondylitis: three cases caused respectively by Pseudomonas aeruginosa, Enterococcus faecalis and Staphylococcus epidermidis. Dystrophin deficient muscular dystrophy in a Labrador Retriever. Emphysematous prostatitis. Erythema multiforme major caused by a Parvovirus infection of keratinocytes. Hemochromatosis due to repeated blood transfusions. Intraspinal synovial cyst. Juvenile nephropathy in the Collie and the Irish Wolfhound. Primary cerebellar cortical degeneration (abiotrophy) in a Scottish terrier. Primary pulmonary artery chondrosarcoma. Renal dysplasia in a Bull Mastiff. Rhabdomyosarcoma (botryoid sarcoma) of the urinary bladder in a Maltese. Spinal mast cell tumor. Spongiform degeneration of the white matter in the central nervous system of Australian Cattle dog. Systemic pasteurellosis caused by Pasteurella canis. Thymic hemorrhage caused by dicumarol intoxication. Undimerized biclonal gammopathy with a single heavy chain class IgA in a dog with multiple myeloma. After a short introduction, the bibliographical data and the abstract of the author(s) and mostly some additional information derived from the article are given. The article will be regularly updated adding overlooked as well as new first reports. PMID- 14535582 TI - First cases of animal diseases published since 2000. 2. Cats. AB - In this second article of a series of papers listing first case reports of animal diseases published since 2000, the following nine cases of cat diseases are discussed: Congenital spongiform change in the brain stem nuclei. Enterococcus hirae enteropathy. Focal cerebral angiomatosis. Glomus tumor. Intraocular extramedullary plasmacytoma. Lens epithelial neoplasias. Phaeohyphomycosis due to Fonsecaea pedrosoi. Pulmonary lymphomatoid granulomatosis. Systemic amyloidosis in a Devon rex. After a short introduction, the bibliographical data, the abstract of the author(s) and some additional information derived from the article are given. The article will be regularly updated adding overlooked as well as new first reports. PMID- 14535583 TI - First cases of animal diseases published since 2000. 3. Cattle. AB - In this third article of a series of papers listing first case reports of animal diseases published since 2000, the following seven cases of cattle diseases are discussed: AL amyloidosis. Canola oil intoxication. Disseminated intracytoplasmic neuronal vacuolation. Encephalomyelitis associated with Akabane virus infection in adult cows. Lower limb deformity: "mirror image duplication of the plantar/palmar half of the distal portion of the digit". Lupinus argenteus intoxication. Novel Propionibacterium infection. After a short introduction, the bibliographical data, the abstract of the author(s), and some additional information derived from the article are given. The article will be regularly updated adding overlooked as well as new first reports. PMID- 14535584 TI - The importance of negative results: methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms in diffuse large B-cell lymphomas (DLBCL). PMID- 14535585 TI - Metallothionein: a multifunctional protein from toxicity to cancer. AB - The metallothionein (MT) family is a class of low molecular weight, intracellular and cysteine-rich proteins presenting high affinity for metal ions. Although the members of this family were discovered nearly 40 years ago, their functional significance remains obscure. Four major MT isoforms, MT-1, MT-2, MT-3 and MT-4, have been identified in mammals. MTs are involved in many pathophysiological processes such as metal ion homeostasis and detoxification, protection against oxidative damage, cell proliferation and apoptosis, chemoresistance and radiotherapy resistance. MT isoforms have been shown to be involved in several aspects of the carcinogenic process, cancer development and progression. MT expression has been implicated as a transient response to any form of stress or injury providing cytoprotective action. Although MT participates in the carcinogenic process, its use as a potential marker of tumor differentiation or cell proliferation, or as a predictor of poor prognosis remains unclear. In the present review the involvement of MT in defense mechanisms to toxicity and in carcinogenicity is discussed. PMID- 14535586 TI - Evaluation of sample handling in relation to levels of tissue inhibitor of metalloproteinases-1 measured in blood by immunoassay. AB - BACKGROUND: The possible effect of preanalytical conditions such as blood sample preparation and handling on TIMP-1 levels in blood needs thorough investigation. MATERIALS AND METHODS: Blood was collected in dry tubes and tubes containing EDTA and kept at 4 degrees C or 20 degrees C for 1, 3, 8, 24 or 72 hours before processing into serum or EDTA plasma. In addition, serum and EDTA plasma samples were frozen and thawed 1-8 times. TIMP-1 was measured by ELISA. RESULTS: Time to processing for up to 72 hours did not significantly affect TIMP-1 levels in serum. In EDTA plasma, TIMP-1 levels were stable for up to eight hours; however, if samples were kept for 24 hours or longer the TIMP-1 levels increased (p < 0.0001). Repeated freezing and thawing had a significant effect on TIMP-1 levels in serum (p = 0.04). In plasma, repeated freezing and thawing for up to six times did not influence TIMP-1. However, in plasma samples exposed to seven or eight freeze/thaw cycles TIMP-1 levels decreased, although not significantly (p = 0.23). CONCLUSIONS: Handling and processing of blood samples is crucial for TIMP 1 measurement by immunoassay. In serum, TIMP-1 levels are unaffected by time to processing. Plasma samples should be processed within eight hours to avoid a TIMP 1 increase. For the measurement of TIMP-1 in archival material, serum should not be used because TIMP-1 levels are significantly affected by repeated freezing and thawing; archival plasma can readily be used provided that samples have not been frozen and thawed more than six times. PMID- 14535587 TI - The value of CEA and CA 19-9 in detecting cancer in a group of high-risk subjects with gastrointestinal symptoms. AB - The aim of this study was to evaluate the clinical value of CEA and CA 19-9 in a potential high-risk population of subjects with gastrointestinal complaints. The basic question was whether the determination of these markers, in addition to some other clinical features in this high risk population, could be helpful in diagnosing intraabdominal cancer. Two hundred and two patients with gastrointestinal complaints underwent standard diagnostic procedures and were followed for at least one year. For every patient, CEA and CA 19-9 levels were obtained at the first examination; the evaluating physician was blinded to the marker levels. The determinants of the likelihood of cancer were evaluated by multivariate analysis. Seventeen patients were diagnosed as having intraabdominal cancers. With the presence of melena (RR = 101.63, p = 0.007), nonspecific gastrointestinal symptoms (RR = 12.54, p = 0.026), increasing age (RR = 1.09, p = 0.028) and abnormal CEA (RR = 240.79, p = 0.000), the risk of having cancer increased significantly and independently. The presence of a primary gastric complaint was associated with a lower risk of cancer in this cohort (RR = 0.01, p = 0.04). Markers were not used in the diagnostic workup. In conclusion, in patients presenting with gastrointestinal complaints, the finding of elevated CEA levels may help in the diagnosis of cancer by prompting a more extensive search for intraabdominal cancer. PMID- 14535588 TI - Assessment of the value of preoperative serum levels of CA 242 and CEA in the staging and postoperative survival of colorectal adenocarcinoma patients. AB - INTRODUCTION: CEA is the most frequently used tumor marker in colorectal cancer. There may be an improvement in its efficacy when used in association with CA 242. AIM: The purpose of this study was to evaluate the efficacy of preoperative serum levels of the tumor markers CA 242 and CEA in the staging and postoperative follow-up of colorectal adenocarcinoma patients. PATIENTS AND METHODS: Of a series of 134 patients with colorectal adenocarcinomas 90 underwent radical surgery and 44 palliative surgery. The control group consisted of 22 organ donors. The cutoff serum levels utilized were 5 ng/mL for CEA and 20 U/mL for CA 242. The mortality during follow-up was recorded in order to determine the duration of survival. The data were submitted to statistical analysis using diagnostic tests, the chi-square test, survival analysis (Kaplan and Meier) and ROC curves. A significance level of p < or = 0.05 was applied. RESULTS: The sensitivity of CEA in Dukes' stages A, B, C and D was 27.8%, 32.4%, 32.1% and 66.7%, respectively. The sensitivity of CA 242 was 11.1%, 16.2%, 30.8% and 50%. When both markers were combined, the sensitivity was 33.3%, 48.6%, 40.7% and 72.5%. In the group of patients who underwent radical surgery the mean survival was 60.47 months for those with high preoperative CEA levels, 52.22 months for those with high preoperative CA 242 levels, and 44.80 months for those with elevated levels of both markers. There was a statistically significant difference in survival between patients undergoing radical surgery with elevated CA 242 levels, especially when CEA was also elevated, and patients without elevated CA 242. CONCLUSION: Preoperative serum levels of CA 242 showed less efficacy than CEA levels for the staging of colorectal adenocarcinoma patients. Elevated preoperative serum levels of CA 242 alone were related to poor survival, especially in association with high levels of CEA. PMID- 14535590 TI - Genetic analysis of H-ras intron-1 polymorphic and variable tandem repeat regions in human breast cancer. AB - H-ras is a member of the ras superfamily of genes. This gene encodes for a 21 kDa protein (p21) which is located on the inner surface of the plasma membrane. Ras genes are involved in a wide variety of human tumors, and there is a known correlation between H-ras activation and breast carcinogenesis. H-ras contains a polymorphic region, a repeated hexanucleotide -GGGCCT - located in intron 1 close to the 5' of the gene (HRM region). Three alleles of this region, P1, P2 and P3, have been identified that contain two, three and four repeats of the hexanucleotide, respectively. H-ras possesses a minisatellite DNA of the variable tandem repeat (VTR) which is located 1000 bp downstream of the gene displaying linkage disequilibrium with HRM. The purpose of this study was to estimate the frequency of P1, P2 and P3 in the normal population and in patients with breast cancer. We studied 56 biopsy specimens from patients with breast cancer, 61 normal blood samples, and 30 pairs of normal and tumoral breast tissues for VTR analysis. There was a difference in the distribution of P1, P2 and P3 alleles between normal and breast cancer samples. The frequency of P1 homozygosity was shown to be almost twice as high in women with breast cancer compared to healthy women (72% versus 39%). These results suggest that P1 homozygosity may be considered as a potential risk factor in breast carcinogenesis. In VTR analysis one sample presented a shift in mobility, but no polymorphism in the BstN I pattern of the 28 bp repetition core was observed. PMID- 14535589 TI - Cytosolic levels of neuron-specific enolase in squamous cell carcinomas of the lung. AB - To study the behavior and possible correlations of neuron-specific enolase (NSE) with other clinicobiological parameters, we measured the cytosolic levels of this marker by means of an immunoradiometric assay (IRMA) in 95 squamous cell lung carcinoma samples. We also analyzed the levels of pS2, tissue-type plasminogen activator (t-PA), hyaluronic acid (HA), free beta subunit of human chorionic gonadotropin (beta-HCG), CYFRA 21.1 and CA 125 in cytosol. On the cell surface we analyzed the concentrations of epidermal growth factor receptor (EGFR), HA, erbB 2 oncoprotein, CD44s, CD44v5 and CD44v6. Other parameters considered were clinical stage, lymph node involvement, histological grade (HG), ploidy and the cellular S-phase fraction measured by flow cytometry on nuclei obtained from fresh tissues. In the 95 squamous cell carcinomas the cytosolic levels of NSE varied from 4.5 to 2235 ng/mg protein (median: 267) and were significantly higher (p < 0.001) than those observed in 38 samples of normal pulmonary tissue obtained from the same patients (range: 56-657; median: 141.5). When classifying tumors according to the different parameters analyzed, we observed that the levels of NSE were higher in aneuploid than in diploid cases (p = 0.046) and in those that were HG3 than in those that were HG2 (p < 0.001). Tumors with high NSE levels (> 422 ng/mg protein; 75th percentile) were more likely to have high S-phase values (p = 0.012) and were more frequently aneuploid (p = 0.038) and HG3 (p < 0.001) than those with low levels of NSE (< 180 ng/mg protein; 25th percentile). These results lead us to the following conclusions: 1) the cytosolic concentrations of NSE are significantly higher in squamous cell carcinomas than in healthy pulmonary tissue, and 2) the cytosolic concentrations of NSE are not correlated with clinical stage or nodal involvement. However, in our study higher levels of the enzyme were statistically correlated with aneuploidy, histological grade 3 and S-phase. This may explain its association with poorer outcome and progression, but also the more favorable response of tumors with elevated NSE to chemotherapy, as suggested by other groups. PMID- 14535591 TI - Epidermal growth factor receptor and c-erbB-2 contents in unresectable (UICC R1 or R2) gastric cancer. AB - BACKGROUND: Epidermal growth factor receptor (EGFR) and c-erbB-2 are membrane receptors expressed in a variety of solid human cancers and directly correlated with poor prognosis. The objective of this work was to evaluate the EGFR and c erbB-2 levels in non-resectable gastric carcinomas, their possible relationship with a variety of clinicopathological tumor parameters, and their prognostic significance. METHODS: This was a prospective analysis of 65 patients with unresectable gastric carcinomas (UICC R1 or R2), who underwent palliative surgery and were followed up for a median period of 13 months. Membranous EGFR levels were examined by radioligand binding assays and cytosolic c-erbB-2 levels by means of an immunoenzymatic assay. RESULTS: There was a wide variability in EGFR (80.3-2910 fmol/mg of protein) and c-erbB-2 (0.4-10071 NHU/mg of protein) levels in neoplastic tissues from patients with unresectable gastric carcinomas. Median c-erbB2 was significantly higher in tumors of the intestinal type than in tumors of the diffuse type (p = 0.035) and in R2 than in R1 tumors (p = 0.016). Statistical analysis showed that there was no relationship between tumor c-erbB-2 or EGFR content and any other patient or tumor characteristics. However, high levels of EGFR were significantly associated with a shorter overall survival (p = 0.01). CONCLUSION: Our data suggest a role of both transmembrane proteins in the progression of gastric cancer. EGFR and c-erbB-2 contents in unresectable gastric cancer could be utilized as appropriate biological markers for selecting candidates for treatment based on EGFR and/or c-erbB-2 inhibition. PMID- 14535593 TI - Methylenetetrahydrofolate reductase genotype in diffuse large B-cell lymphomas with and without hypermethylation of the DNA repair gene O6-methylguanine DNA methyltransferase. AB - C677T and A1298C methylenetetrahydrofolate reductase (MTHFR) polymorphisms have been suggested to affect susceptibility to malignant lymphoma, possibly by altering DNA methylation. The DNA repair gene O6-methylguanine DNA methyltransferase (MGMT) is transcriptionally silenced by promoter hypermethylation in diffuse large B-cell lymphomas (DLBCL). We analyzed the MTHFR677 and MTHFR1298 genotypes in 111 DLBCL patients and 465 controls. No significant difference in the frequency of MTHFR polymorphisms between patients and controls and no significant association between MTHFR677 or MTHFR1298 genotypes and methylation of MGMT promoter were observed. These results indicate that MTHFR variants are not related to DLBCL development and MGMT hypermethylation. PMID- 14535592 TI - Limited prognostic value of c-erbB-2 compared to uPA and PAI-1 in primary breast carcinoma. AB - In a retrospective study of 488 women with primary breast cancer, after a median follow-up of 10 years, we sought interactions between disease-free survival (DFS) and overall survival (OS) and tumor antigen levels of two components of the plasminogen system, urokinase-type plasminogen activator (uPA) and its inhibitor PAI-1, and the transmembrane growth factor receptor c-erbB-2. We used ELISAs (American Diagnostica, Greenwich, CT, USA) to quantify uPA and PAI-1 antigen levels in cytosols, and a double monoclonal antibody-based assay (EIA) (Ciba Corning Diagnostics, Alameda, CA, USA) to quantify c-erbB-2 in membrane extracts of the same tissues. Weak positive correlations were found between uPA and c-erbB 2 (r(s) = 0.146; p = 0.001) and between PAI-1 and c-erbB-2 (r(s) = 0.154; p < 0.001). In the overall population, using univariate analyses, c-erbB-2 overexpression and high uPA and PAI-1 antigen levels (> 300 IU/mg, > 1.40 ng/mg and > 5.53 ng/mg, respectively) were significantly associated with shorter DFS (p = 0.003, p < 0.001 and p < 0.001, respectively) and OS (p < 0.001 in all cases). Using multivariate analyses, PAI-1, node status and tumor size were independent predictors of DFS and c-erbB-2 was retained in the model only for OS. In the node negative subgroup, PAI-1 was the strongest significant survival predictor both for OS (p = 0.003; HR 2.52) and DFS (p < 0.001; HR 2.39). This study shows that in primary breast cancer c-erbB-2 offers no additional prognostic information when uPA and/or PAI-1 are candidates in the multivariate analyses. PMID- 14535594 TI - Are 90K/MAC-2BP serum levels correlated with poor prognosis in HCC patients? Preliminary results. AB - In this study we assessed the prognostic significance of 90K/MAC-2BP serum levels in a group of 40 hepatocellular carcinoma patients. This glycoprotein is a new, interesting serum marker that reflects the immune reaction of the host against certain viral infections and tumors such as breast, ovarian and pancreatic cancer. Hepatocellular carcinoma (HCC) is one of the most widespread tumors in the world. AFP is currently the most useful marker for HCC, in spite of its poor diagnostic sensitivity. In this study 40 cirrhotic HCC patients were enrolled. The prevalence of viral hepatic infections in this group was 73% for HCV, 8% for HBV, and 8% for both viruses. Thirteen percent of the patients showed non-virus related liver damage. 90K serum levels were assayed by an ELISA kit and AFP levels by a chemiluminescent enzyme immunometric system. The overall survival curves were estimated by the Kaplan-Meier method, taking into account age, sex, 90K and AFP serum levels. Statistical analysis showed a highly significant influence on overall survival of age below 70 years and 90K serum levels below the cutoff of 14 ng/mL. Serum AFP (< or = 20 ng/mL) had positive prognostic value only when it was associated with 90K levels (p < 0.02, log-rank). PMID- 14535595 TI - ROC curves--can differences in AUCs be significant? PMID- 14535596 TI - Interchangeability between control material and patient serum in tumor biomarker assessment. PMID- 14535597 TI - Racial/ethnic differences in the self-reported use of screening mammography. AB - The efficacy of mammography in reducing breast cancer mortality among women 50-69 years of age has been demonstrated in randomized controlled studies, but many women, especially ethnic minorities, have not been receiving regular mammographic screening. The current study investigated racial/ethnic differences in mammography use and their association with demographic characteristics and other factors. The study population consisted of 4,444 women aged 40 years and older who participated in the 1996 Medical Expenditure Panel Survey. Outcome measures studied included the self-reporting of mammography within the past two years and past year. Multivariate logistic regression modeling was used to examine the effect of race while controlling for other factors. In the univariate analysis, there was virtually no difference between white, black, and Hispanic women in mammography rates within either one or two years. However, multivariate logistic regression suggested that both blacks and Hispanics were more likely than whites to have received recent mammography, as black women were 31% and Hispanic women were 43% more likely than white women to have had a mammogram within the previous two years. Our results suggest that white women are no longer more likely to receive periodic screening mammography than black and Hispanic women, and in fact, might even be less likely to undergo the procedure. This reversal might indicate, at least in part, that programs and other activities to promote screening mammography among ethnic minority women have been successful and should now be expanded to include other women. PMID- 14535599 TI - Validation of birth certificate data in New York State. AB - Vital statistics birth certificate data are an important source of information for researchers, policy makers, and state officials to evaluate the quality of care delivered to pregnant women. The purpose of this study was to assess the validity of data elements being reported by the hospitals on the birth certificate record when compared to the medical record. This study used a random sample of birth certificates from two upstate and two downstate counties, in New York State, comprising a total of 100 records per county. The review assessed data elements from seven major categories: prenatal care, maternal medical risk factors, risk factors related to pregnancy, lifestyle risk factors, method of delivery, complications of labor and delivery, and infant information. Sensitivity, specificity, the positive predictive value and the negative predictive value assessed level of agreement between the birth certificate and the medical record. Overall, the birth certificate data reflected high specificity, because most conditions are rare. The sensitivity of the data was more varied, ranging from 0 to 100%, reflecting that if a rare condition was present it often was not documented on the birth certificate. Many of the data elements are reported accurately. However, caution should be used for data elements that are poorly reported. PMID- 14535598 TI - Healthcare access and utilization among women 40 and older at the U.S.-Mexico border: predictors of a routine check-up. AB - Mexican Americans are more likely to experience barriers to access and utilization of healthcare services than any other U.S. Hispanic group. In Mexico, where the majority of the population has access to care, the pressing issue is the underutilization of preventive services among adults. This study was conducted to assess access and utilization barriers among a U.S.-Mexico border population. A cross-sectional, population-based survey was conducted during 1999 2000 in a pair of contiguous U.S.-Mexico border communities. Household surveys were administered to U.S. and Mexican women, 40 years of age and older, to assess healthcare access and utilization, participation in chronic disease screenings, orientation toward prevention and personal history of chronic disease. Analysis indicates few statistically significant differences (p < 0.05) among access and utilization variables by country. Mexican participants were more likely to have a regular source of care and to have had a blood sugar test within the past 12 months. U.S. participants more often reported having had a Pap smear and mammogram during the previous year. Factors independently positively associated with having had a routine check-up during the past 12 months included age and having a regular provider or place to go when sick. Only going to the doctor when ill was independently inversely associated with routine check-ups in the past 12 months. Findings suggest that U.S. and Mexican border populations are similar with regard to healthcare access and utilization characteristics. Efforts to increase utilization of preventive health screenings among women are needed at the U.S.-Mexico border. PMID- 14535600 TI - Using an integrated approach to understand vaccination behavior among young men who have sex with men: stages of change, the health belief model, and self efficacy. AB - Studies continue to show that the majority of men who have sex with men (MSM) in the United States remain unvaccinated against hepatitis A (HAV); such limited vaccination coverage is a missed opportunity for preventing disease. This study sought to identify beliefs and attitudes associated with motivational readiness for vaccination against HAV among MSM, using a theoretically-integrated framework. Questionnaire data were collected from 358 MSM through bar outreach. MSM with increased readiness to complete the two-dose series perceived lower practical barriers and reported greater healthcare provider communication about their sexual orientation and risk behavior. They also perceived higher benefits to vaccination and higher severity of infection, and had higher self-efficacy to complete the vaccine series. Relationships between stages of change and theory based constructs from the health belief model and the social learning theory follow predicted patterns suggesting that these theories may provide useful frameworks for understanding vaccination readiness and intervention strategy development among MSM. PMID- 14535601 TI - Local newspapers, community partnerships, and health improvement projects: their roles in a comprehensive community initiative. AB - To understand local media's role in a community health initiative, a content analysis of 1,709 paragraphs from 173 news articles and editorials was undertaken. The articles were from three local newspapers, one in each of three communities. Analyses focused on article content combined with reflective personal interviews with local campaign directors. Results suggest that local campaign staff can be successful using commercial media to achieve objectives. Surprisingly, most coverage was not about projects with observable and easily identifiable benefits for local residents, but rather partnerships among influential residents engaged in decision-making about such projects. We conclude that the politics of resource distribution is more newsworthy to local journalists than tangible topics like access to health information, insurance coverage, and service provision. PMID- 14535602 TI - Osteoporosis prevention: knowledge and behavior in a southwestern community. AB - Prior to developing an osteoporosis prevention education program and social marketing campaign, we sought to (a) establish current status of osteoporosis related knowledge and behavior among women aged 25-55 years in Maricopa County, Arizona, and (b) assess factors that segment the population by age and ethnicity. Two-hundred women were surveyed by telephone using random-digit dialing selection. Data demonstrated knowledge of need to consume adequate calcium, but mixed understanding of forms of exercise that help prevent osteoporosis. Knowledge of osteoporosis prevention did not differ as a function of menopause status. Differences for Hispanic versus non-Hispanic women's knowledge showed fewer correct responses for Hispanics for dietary and physical activity questions, and more correct responses on the relationship between body weight and osteoporosis risk. Hispanic women and post-menopausal women generally fit the pattern of higher risk behaviors with Hispanic women exercising and using HRT less and postmenopausal women reporting lower calcium intake and physical activity and more tobacco use. Hispanic women appeared to have similar intake of dietary calcium despite lower levels of milk products. Social marketing campaigns for osteoporosis prevention should be segmented for cultural and age differences, especially considering differences in orientations toward exercise, milk consumption, and competing emphasis on other diseases. PMID- 14535603 TI - Age and the gender gap in distress. AB - Women report more psychological distress than men and recent evidence suggests that this gap increases with age. It has been argued that the widening differential in distress reflects the progressive and cumulative nature of women's disadvantaged work and family roles. Drawing on the cumulative disadvantage hypothesis and social stress theory, we test: (1) whether exposure to chronic stress accounts for an increasingly larger proportion of the gender effect on distress with age; and (2) whether women are increasingly more vulnerable to the effects of chronic stress on distress with age. Data are from the 1994 wave of the Canadian National Population Health Survey, a national probability sample of women and men aged 20 and older (N = 13,798). Exposure to long-term stress helps us understand gender differences in distress for those in their pre-retirement years. However, contrary to the cumulative disadvantage hypothesis, the model became increasingly less likely to explain such differences with age. Gendered vulnerability to long-term stress was not evident in the sample. The implications of these findings are discussed with particular reference to our ongoing efforts to understand health in the context of social structure and subjectivity. PMID- 14535604 TI - Women's satisfaction with breast and gynecological cancer screening. AB - PURPOSES: This study examined women's satisfaction with their breast and gynecological (GYN) cancer screening, clinical breast examination (CBE), mammogram (MAM), and pelvic examination (PE). Selected predictors (characteristics of the exam and characteristics of the health care provider) of satisfaction with their screening exams also were assessed. METHODS: Women (N = 242) ages 19 years and older were recruited from a residential mailing list, and from church and community organizations. Participants completed questionnaires in their homes or in the community setting where they had been recruited. Descriptive statistics were used to describe the level of women's satisfaction with their most recent exams. Hierarchical multiple regression was used to determine the influence of exam and health care provider characteristics on women's satisfaction with exams. RESULTS: We found that women were very satisfied with care received during all three screening exams. Women were more satisfied during CBE and PE if they perceived these exams as informative, clear and complete and if they perceived the provider as informative and responsive to them when they ask questions. Women also were more satisfied with all three exams when the provider was perceived as relaxed during these exams. PMID- 14535605 TI - Do main partner conflict, power dynamics, and control over use of male condoms predict subsequent use of the female condom? AB - This study assessed hypotheses that measures of power and control over male condom (MC) use would predict use of the female condom (FC) among women with main partners from two public STD clinics (n = 616). The women (mean age 24 years, 87% African American) were enrolled in an intervention study to promote barrier contraceptive use and were interviewed at baseline and at 6 monthly follow-up visits. Seven baseline predictor variables were assessed: her having requested MC use, his having objected, her having wanted a MC used but not asking, percentage of MC use, perceived control over MC use, anticipated consequences of refusing unprotected sex, and physical violence. In the first Poisson regression analysis, none of the hypothesized predictors was significantly associated with FC use during follow up. In the second regression analysis, which assessed the influence of the hypothesized set of predictors on follow-up FC use in situations when MCs were not used, we found two effects. Either no or inconsistent MC use before study entry was associated with less subsequent FC use; women who reported, at study entry, having more control over MC use were more likely to use FCs during follow up. We found no evidence of adoption of the FC by women in relationships marked by history of conflict over the MC, circumstances in which alternatives are most needed. On the contrary, we found that women with a history of control and consistent use of MCs were the most likely users of FCs when MCs were not used. PMID- 14535606 TI - Women's hysterectomy experiences and decision-making. AB - The goal of the study was to examine women's experiences with gynaecologic symptoms and how they decided to undergo hysterectomy. For this purpose, twenty nine women were interviewed in hospital within three days of undergoing hysterectomy. The interviews elicited information about the nature of the problem that caused the women to seek medical help, actions taken to solve their problem, their relationship with their gynaecologist, information seeking patterns and decision-making about hysterectomy. Although findings revealed that the symptoms women suffered had a negative impact on their lives, most women delayed seeking medical help and attributed their symptoms to factors other than a physical problem in their reproductive system. Most of the participants' information about the symptoms and possible treatments came from their consulting other women with similar problems. The women reported that their gynaecologist did not initiate a comprehensive discussion about other treatments and their advantages and disadvantages. Only women who had informed themselves about other treatments actively discussed alternatives to hysterectomy with their physicians. The women's decision-making process about undergoing hysterectomy was difficult and depended primarily on the women's illness experiences, age, wish for future children, information they gathered from their gynaecologist and from other women. The findings are discussed in relation to the importance of information provision by gynaecologists and its effects on women's decision-making about hysterectomy. PMID- 14535607 TI - Investigating Hong Kong's Filipino domestic workers' healthcare behavior, knowledge, beliefs and attitudes towards cervical cancer and cervical screening. AB - The aim of this study was to investigate Hong Kong Filipino domestic workers health behaviors, knowledge, beliefs and attitudes about cervical screening and cancer. A concurrent cross sectional survey design used a snowball method of recruitment was used because of the acknowledged problematic access to a random sample of immigrant women. A total of 98 female domestic helpers were actively recruited through designated recreation centers. The women were between 24-45 years old (mean = 37.9, SD = 7.7). The majority of these women were employed as full-time domestic helpers (91%), were earning less than dollar 4000 Hong Kong dollars/month (92%), were married (82%), with children (91%), were non-smokers (88%), and had at least a secondary level of education (100%), with 66% of these women having completed post-secondary education. While the majority of women had previously heard about cervical smears (78%) more than half (53%) reported never having a cervical smear taken. Women who had a prior cervical smear had significantly more knowledge about cervical smears and cervical cancer (mean = 51.34, SD = 2.5) than those who never had a cervical smear (mean = 49.72, SD = 3.2). Recommendations are made for culturally tailored mass screening programmers out of office hours and health information to be provided in both written and oral Tagalong. Doctors and nurse ideally of Filipino origin should be used to deliver health messages that prioritize the importance of self protection for the family and future fertility issues, culturally prized within Filipino society. PMID- 14535608 TI - Physical activity programs for refugee Somali women: working out in a new country. AB - Islamic refugee women from non-westernized countries face a number of challenges in adapting to their new country, especially when that new country is westernized and is not Islamic. Refugees are primarily women and children, so it is important that women be in their best health because they usually bear the responsibility of caring for each other and children, often in very difficult situations. Maintaining or obtaining good levels of physical activity contributes to good health: mentally, physically and socially. At the request of women in the local Somali community, a number of initiatives were taken to increase their opportunities for physical activity. Through interviews, observations and conversations we explored barriers to fitness and exercise, the social, physical, and cultural effects of physical activity, and solutions to facilitate Somali women's access to fitness and exercise opportunities. Physical activity interventions included exercise classes in a community center used by the Somali community, trial memberships at a local women-only fitness center, and walking and sports groups. We discuss the procedural issues relating to setting-up these physical activity opportunities, the results of interviews with 37 of the women about their health and perceptions and issues relating to the physical activity options, and our recommendations for setting up similar classes with other Somali or Islamic communities. PMID- 14535609 TI - Medicalization vs. adaptive models? Sense-making in magazine framing of menopause. AB - Women over the age of 40 are largely absent from media imagery. This study examined how magazines have framed menopause over the past two decades, as an increasing number of women have entered the phase. Using a traditional content analysis, we analyzed author and source gender, topics, and photographs in menopause articles in seven news and women's magazines. A computerized content analysis program was used to determine how the magazines framed menopause. "Framing" refers to the emphasis and theme(s) of an article. Women's magazines provided a broad range of topics to help women prepare and cope; news magazines reported scientific developments, particularly in fertility. More frames, including clearer descriptions of symptoms and effects, occurred in women's magazines. Female authors included menopausal women as sources, but males did not. Photos of menopausal women appeared in a small portion of articles, and the women shown were predominantly white. Findings indicate some consistency with media coverage of other women's health issues, but with greater differences between news and women's magazines. PMID- 14535610 TI - FRONTLINE-the suicide of patients and the quiet voice of the therapist. PMID- 14535611 TI - Terminal mental illness: resident experience of patient suicide. AB - Suicide prevention efforts are a major focus of psychiatry residency training. Residents are taught to identify suicide risk factors, monitor for suicidal ideation, and develop crisis stabilization plans for patients at risk for self harm. In contrast, training and support for dealing with suicide completion is often lacking. Although suicide remains a predictable outcome for many patients with severe mental illness, this topic may be avoided or reviewed only with residents who are directly affected by patient suicide. The purpose of this paper is to present a psychiatry resident's experience of dealing with a patient suicide and identify obstacles to developing this aspect of training. Options for "preventative" training in helping trainees deal with this unfortunate outcome of mental illness will be reviewed. PMID- 14535612 TI - When depression becomes terminal: the impact of patient suicide during residency. AB - Patient suicide often results in profound personal and professional crises for the treating clinicians. Residency training in psychiatry represents a critical time for experiencing the death of patients by suicide. While residents often treat some of the most acutely ill, high-risk patients, during a uniquely vulnerable phase of professional development, many training programs do not have formalized procedures in place for aiding residents in the event of patient suicide. The working through of a suicide offers an opportunity for personal and professional growth. This paper examines this issue from the perspective of a resident-in-training who experienced the suicide of a long-term outpatient. Issues explored include personal and professional reactions to the patient's death as well as a discussion of the organizational response to the event. PMID- 14535613 TI - When a psychiatry resident's patient commits suicide: transference trials and tribulations. AB - It is an unfortunate reality that a substantial proportion of psychiatry residents will experience the suicide death of one or more of their patients during the course of their training. The psychological impact of such deaths may result in marked repercussions on the treating resident; but, in addition, many other individuals and entities--resident peers, supervisors, other mental health professionals and staff, training directors, training programs, psychiatry departments--may be affected as well to a greater or lesser extent. This paper explores one manifestation of the impact on a psychiatry resident of a patient's suicide death: the activation of transference reactions and transference enactments on the part of both a resident and a residency training director. The author argues that the very breadth and depth of such transference responses offer invaluable opportunities for self-observation, psychological introspection, and personal growth if involved individuals have sufficient courage and supportive mechanisms for processing of the patient suicide and its aftermath. The failure of many current psychiatric trainees to undergo personal intensive psychotherapy is identified as a major obstacle to turning such potentially destructive experiences into learning and growth opportunities. PMID- 14535615 TI - Object relations in Harry Potter. AB - Good fiction helps children address their emotional dilemmas by evoking repressed content, and offering strategies and meaningful values that help them work towards resolutions. Because certain fundamental conflicts continue to be revisited and reworked throughout adulthood, it follows that masterful children's literature might enthrall adults as well. Given the extraordinary, worldwide success of the Harry Potter stories with both children and adults, it might be inferred that they, indeed, are among such literature. Common object relations themes, as well as other intrapsychic processes, are presented in such an imaginative and resonant way that the unconscious is readily engaged. The character of Harry Potter, specifically, embodies such universal (repressed) torments as the agony of destroying and losing the mother; the ominous perception of good and bad objects at war within the self; and the earnest reparative efforts offered to save the self from eternal separation from the beloved other. PMID- 14535614 TI - Somatization and conversion: distinct or overlapping constructs? AB - The terms somatization and conversion are used descriptively to define specific diagnostic entities, and phenomenologically to denote pathologic processes that underlie somatic symptom formation. There is a lack of clarity, and even controversy, however, as to whether somatization and conversion should be considered distinct or overlapping constructs, and whether they contribute to the pathogenesis of certain organic diseases or solely to medically unexplained somatic symptoms. This article attempts to resolve some of this confusion by reviewing the origins of the terms somatization and conversion, and describing how their meanings and uses have evolved over the last century. Whereas some psychoanalysts and psychiatrists adopted Stekel's view that somatization involves a psychological process analogous to conversion, others maintained Freud's distinction between the psychoneuroses and the actual neuroses and view somatization as a physiological process. Recent psychoanalytic attempts to understand somatization are based on a modern theory of emotional processing, which is rooted in cognitive science and open to empirical research. Maintaining a conceptual distinction between somatization and conversion has important implications for psychoanalytic therapy. PMID- 14535616 TI - Assaultive projective identification and the plundering of the victim's identity. AB - A destructive mode of projective identification is delineated: a predator's catastrophic attack calculated to cause the victim a stress disorder marked by a disarrayed identity. This discomposure enables the perpetrator to aggrandize a manifold inroad upon the victim's identity to imprint, intrude, mingle and/or lodge his representation into it; to ravage, steal from, impoverish, and/or corrupt it; finally to have his representation emerge as an internal regulator of the traumatized prey's functioning. The victim's debased integrity is manifest in the symptoms of his ensuing posttraumatic stress disorder (PTSD). PMID- 14535617 TI - Psychoanalytic peregrination VI: "the effect on countertransference of the collapse of civilization". AB - There is an aspect to countertransference that has not been discussed very much in the literature of psychoanalysis, one that is becoming increasingly important. This centers around the effect on the analyst's countertransference of the deterioration of the culture in which he or she is immersed. I discuss the decline of Western civilization as it is described by a number of contemporary authors in order to show how changes in the value system of the psychoanalyst or his or her turning a blind eye to the injustices and barbarism of the cultural surround both affect the capacity to immerse in the creative process of psychoanalysis, and send a tacit message to the patient about the permissability of hiding certain material. All this contradicts the "fundamental rule" of psychoanalysis, and disturbs the analyst's listening process. The impact of the dumbing down and deterioration of the educational, cultural, and home upbringing milieu of the new generation of psychoanalysts will be profound in influencing the future practice of psychoanalysis. PMID- 14535618 TI - The rape of boys and the impact of sexually predatory environments: review and case reports. AB - This paper describes the effect of rape and the impact of sexually predatory environments on young boys and the resulting sequelae in adulthood. It presents the literature and the case histories of Jeff and Todd to illustrate the psychic devastation that occurs, and the ensuing attempts at reestablishing boundaries and regaining a sense of autonomy and male identity. Issues including facing powerlessness, betrayal by trusted adults, and the effect of emasculation on developing sexual identity are explored. A study of almost 4,000 siblings by Nelson et al. (2002) showed that a twin who was not sexually abused himself but who grew up in the same milieu as his sexually abused twin, suffered similar psychopathologic symptoms, such as suicide attempts, alcohol dependence, and social anxiety. Todd spent his formative years in a Catholic boarding school where boys were physically and sexually abused by priests. His history illustrates the severe psychological damage this causes and is consistent with the findings of Nelson's study. PMID- 14535619 TI - The aim of my life is to study the nature. PMID- 14535620 TI - Kappa-receptor selective binding of opioid ligands with a heterocyclic bicyclo[3.3.1]nonan-9-one structure. AB - Previous pharmacological results have suggested that members of the heterocyclic bicyclo[3.3.1]nonan-9-one-like compounds are potent kappa-opioid receptor specific agonists. One lead molecule of this series. called compound 1 (dimethyl 7-methyl-2,4-di-2-pyridyl-3.7-diazabicyclo[3.3.1]nonan-9-one-1,5-dicarboxylate) exhibited high affinity for [3H]ethylketocyclazocine and [3H]U-69.593 binding sites in guinea pig cerebellar membranes which known to be a good source for kappa1 receptors. It was shown by molecular modelling that heterocyclic bicyclo[3.3.1]nonan-9-ones fit very well with the structure of ketazocine, a prototypic kappa-selective benzomorphan compound; when compared to the arylacetamide structure of U-69.593, a specific kappa1-receptor agonist, a similar geometry was found with a slightly different distribution of the charges. It is postulated, that the essential structural skeleton involved in the opioid activity is an aryl-propyl-amine element distributed along the N7-C6-C5-C4-aryl bonds. PMID- 14535621 TI - Upregulation of mu opioid receptors by voluntary morphine administration in drinking water. AB - Morphine was provided to rats in drinking water for 21 days. Profound analgesic tolerance was detected both in hot-plate and tail-flick tests. The density of [3H]DAMGO binding sites increased by 76% in spinal cord membranes due to morphine exposure compared to those in opioid naive animals. Slightly augmented [3H]DAMGO binding was measured in the synaptic plasma membranes, with a concomitant decrease in the microsomal membranes, of morphine tolerant/dependent brains. These observations suggest that the regulation of spinal mu opioid receptors might be different from those in the brain. It is emphasized that the molecular changes underlying tolerance/dependence are influenced by several factors, such as the tissue or subcellular fractions used, besides the obvious importance of the route of drug administration. Results obtained after voluntary morphine intake further support the growing number of experimental data that chronic morphine does not internalize/downregulate the mu opioid receptors in the central nervous system. PMID- 14535622 TI - Restraint stress and anti-tumor immune response in mice. AB - Psychological stress modulates the immune system through the hypothalamic pituitary-adrenal axis, the sympatho-adrenomedullary axis and the opioid system. According to literature data, restraint stress increases the immune cell apoptosis, decreases the spleen and thymus cell content, the natural killer (NK) activity in the spleen, and it compromises the anti-tumor immune response in mice. We immobilized mice in two consecutive nights, and then determined the cell number, apoptosis, NK cell content, NK activity and the level of cytokine mRNAs (TNF-beta, TNF-alpha, IL-4, IL-5, IL-1alpha, IFN-gamma, IL-2, IL-6, IL-1beta and IL-3) in the thymus and spleen. No consistent changes were detected in any of the immune parameters either in C57Bl/6 or in DBA/2 mice. Stressed or control B6 mice were injected with B16 melanoma cells immediately after the immobilization or one week later. No significant differences were found in the growth of primary tumors and lung metastases in stressed and control animals. Taken together, our mice, kept in a general-purpose non-SPF animal house, seemed to be refractory to the stress-induced immunomodulation. Our interpretation is that stress-induced immunomodulation can occur only in mice isolated from any background stressors, or rather natural stimuli, throughout their life. PMID- 14535623 TI - Effects of structural modifications of N-CPM-normorphine derivatives on agonist and antagonist activities in isolated organs. AB - The agonistic and antagonistic properties of N-cyclopropylmethyl (N-CPM) morphine derivatives were observed in mouse vas deferens (MVD), longitudinal muscle of guinea pig ileum (GPI) and rabbit vas deferens (LVD). In MVD the K(e) values of the titled compounds (N-CPM-morphine, N-CPM-isomorphine, N-CPM-dihydromorphine, N CPM-dihydroisomorpPhine, N-CPM-dihydromorphone and naltrexone) were measured for mu-, kappa- and delta-receptors using normorphine, ethylketocyclazocine (EKC) and D-Pen2-D-Pen5-enkephaline (DPDPE) as selective agonists on the receptors, respectively. For mu-receptors of MVD the tested compounds showed similar affinity. For kappa-receptors the non-iso-6-OH derivatives possessed much less affinity than the iso-derivatives. Similar difference could be observed for delta receptors. The agonistic activities of these compounds in MVD were observed to be between 0-20% of the inhibition of muscle contractions. In GPI the compounds except naltrexone possessed strong agonistic activities effectively antagonized by nor-binaltorphimine (nor-BNI) (K(e) of nor-BNI was 0.23 nM) suggesting that they were strong kappa-receptor agonists. We investigated these agents in LVD too, which contains kappa-receptors, but they did not produce any agonist potencies. It raises the possibility that the kappa-receptor subtypes of LVD and MVD are different from the kappa-receptor subtype of GPI or the vasa deferentia contain much fewer kappa-receptors than GPI and the intrinsic activities of these compounds are too small to reach the 50% inhibition of the contractions. PMID- 14535625 TI - The P2Y nucleotide receptors in the human genome. AB - Since the first identification of P2Y receptor sequences in 1993, it has quickly become apparent that this family of the G-protein coupled receptors is very diverse. Members of this receptor family are activated extra-cellularly by a wide variety of adenosine and uridine nucleotides including sugar-nucleotides. The recent decipherment of the Human Genome has enabled us to search for new, yet undiscovered P2Y receptor subtypes. In this article we examine the relationships of six orphan G-protein coupled receptor (GPCR) sequences which show considerable sequence homology to various P2Y receptors. The clustering at a few chromosomal loci of P2Y receptor genes and their related orphan genes further suggests that particular P2Y subsets were derived from the same ancestral gene during evolution. PMID- 14535624 TI - Potencies and selectivities of inhibitors of acetylcholinesterase and its molecular forms in normal and Alzheimer's disease brain. AB - Eight inhibitors of acetylcholinesterase (AChE), tacrine, bis-tacrine, donepezil, rivastigmine, galantamine, heptyl-physostigmine, TAK-147 and metrifonate, were compared with regard to their effects on AChE and butyrylcholinesterase (BuChE) in normal human brain cortex. Additionally, the IC50 values of different molecular forms of AChE (monomeric, G1, and tetrameric, G4) were determined in the cerebral cortex in both normal and Alzheimer's human brains. The most selective AChE inhibitors, in decreasing sequence, were in order: TAK-147, donepezil and galantamine. For BuChE, the most specific was rivastigmine. However, none of these inhibitors was absolutely specific for AChE or BuChE. Among these inhibitors, tacrine, bis-tacrine, TAK-147, metrifonate and galantamine inhibited both the G1 and G4 AChE forms equally well. Interestingly, the AChE molecular forms in Alzheimer samples were more sensitive to some of the inhibitors as compared with the normal samples. Only one inhibitor, rivastigmine, displayed preferential inhibition for the G1 form of AChE. We conclude that a molecular form-specific inhibitor may have therapeutic applications in inhibiting the G1 form, which is relatively unchanged in Alzheimer's brain. PMID- 14535626 TI - The molecular mechanisms of cellular tolerance to delta-opioid agonists. A minireview. AB - Chronic treatment with deltaopioid agonists, similar to other agonist drugs, causes tolerance. Tolerance is a complex adaptation process that consists of multiple, cellular and neural-system adaptations. Cellular tolerance to delta opioid agonists involves feedback-regulation of the function, concentration, and localization of the delta-opioid receptors (receptor desensitization) as well as of intracellular effectors (functional desensitization). We are using a recombinant Chinese hamster ovary cell line expressing the human delta-opioid receptors (hDOR/CHO) to investigate the molecular mechanisms of cellular tolerance. We found that the structurally distinct delta-opioid agonists mediate receptor down-regulation by different mechanisms. Thus, truncation of the last 35 C-terminal amino acids of the hDOR completely abolished DPDPE, but not SNC 80 mediated receptor down-regulation. In addition, down-regulation of the wild type , and the truncated hDORs exhibited different inhibitor sensitivity-profile. Chronic delta-opioid agonist treatment also causes functional desensitization of forskolin-stimulated cAMP formation and cAMP overshoot in the hDOR/CHO cells. We have demonstrated that chronic SNC 80 treatment also causes concurrent phosphorylation of the adenylyl cyclase (AC) VI isoenzyme hDOR/CHO cells. Both AC superactivation and AC VI phosphorylation were SNC 80 dose-dependent, naltrindole sensitive, and exhibited similar time course-, and protein kinase inhibitor sensitivity profile. We hypothesize that phosphorylation of AC VI plays an important role in delta-opioid agonist-mediated AC superactivation in hDOR/CHO cells. PMID- 14535627 TI - The relation between serum leptin levels and body fat mass in patients with active lung tuberculosis. AB - The relationship of leptin to diminished appetite and weight loss has been investigated in many diseases. Diminished appetite and weight loss are the most apparent characteristics of patients with active lung tuberculosis and in this study the relation of leptin to such diminished appetite and weight loss has been investigated in patients with active lung tuberculosis before and after treatment. Twenty-five patients (7 female, 18 male) with active tuberculosis having an age range of 18-70 years (mean 47.48 +/- 15.36 y) and 25 normal individuals (9 female, 16 male) having an age range of 25-71 years (mean 44.60 +/ 13.80 y) were included in this study. Leptin levels, body mass index (BMI), body fat ratio (BFR), and waist hip ratio (WHR) were measured before and after 6 months of antituberculosis treatment. The same measurements were also made in the control group and the results were compared. While the pretreatment BMI (22.02 +/ 4.31 kg/m2) and BFR (16.60% +/- 9.30%) values in the patient group were significantly lower than in the control group, we found no difference in their pretreatment WHR values. Pretreatment leptin levels (3.49 +/- 3.34 microg/L) were significantly higher in patients with tuberculosis than in the control group (2.33 +/- 1.10 microg/L). Leptin levels were found to be significantly increased at the 6th month of antituberculosis treatment (5.65 +/- 5.41 microg/L) than the pretreatment values (p < 0.05). We observed an evident increase in BMI (24.10 +/- 4.87 kg/m2) and BFR (17.51% +/- 9.25%) due to antituberculosis treatment (p < 0.05). This study suggests that leptin has a role in the diminished appetite and weight loss symptoms in patients with active lung tuberculosis. PMID- 14535628 TI - No effect of simvastatin treatment on insulin sensitivity in patients with primary hypercholesterolemia. AB - OBJECTIVE: Statins, in addition to cholesterol lowering, have nonlipid effects on formation and progression of atheromatous plaque. Insulin resistance and hyperinsulinemia may have detrimental influences on the arterial wall. Statins (may also) inhibit insulin signal transferring in vascular smooth cell cultures. However, their effect on insulin sensitivity remains controversial. Therefore, we decided to investigate the effect of simvastatin on insulin sensitivity in hypercholesterolemic patients. PATIENTS AND METHODS: Eighteen patients with primary hypercholesterolemia were divided into simvastatin group (n = 9; 4 females, 5 males; BMI 30.6 +/- 4 kg/m2; mean ages 57 +/- 6 years) and placebo group (n = 9; 4 females, 5 males; BMI 28 +/- 2.9 kg/m2; mean ages 49 +/- 10 years). Simvastatin (20 mg/day) or placebo were given for 2 months. Total and HDL cholesterol were measured and LDL cholesterol was calculated by Friedewald formula. Insulin sensitivity was determined by using euglycemic hyperinsulinemic clamp technique [40 microU/m2/min insulin infusion rate; glucose disposal rate (M)= mg/kg/min] before and after treatment. RESULTS: Plasma levels of total, LDL and HDL cholesterol decreased significantly in simvastatin group after treatment (p = 0.000, p = 0.000, and p = 0.048, respectively). Plasma levels of total cholesterol decreased significantly (p = 0.032), whereas LDL and HDL levels remained unchanged in placebo group. M value (mg/kg/min) decreased insignificantly in simvastatin group (4.32 +/- 1.57 vs. 3.71 +/- 1.91) and increased in placebo group (3.55 +/- 1.91 vs. 3.95 +/- 0.95). CONCLUSION: Short term simvastatin treatment did not affect insulin sensitivity determined by "gold standard" euglycemic hyperinsulinemic clamp method in hypercholesterolemic patients in this research. Further studies with simvastatin using higher doses and longer duration should be performed. PMID- 14535629 TI - Expression of cAMP response element binding protein (CREB)-binding protein (CBP) and the implication in retinoic acid-inducible transcription activation in human salivary gland adenocarcinoma cell line HSG. AB - In the process of retinoic acid (RA) signaling, retinoic acid receptor interacts with a coactivator complex composed of various transcription cofactors such as CREB-binding protein (CBP)/p300 and p160 family member proteins represented by steroid receptor coactivator-1 (SRC-1)/NCoA1 and p300/CBP cointegrator protein (p/CIP)/ACTR. In order to investigate the relationship of CBP to the RA signaling in a human salivary gland (HSG) adenocarcinoma cell line, we examined the expression of CBP in the cells. Immunoprecipitation and immunoblotting of the nuclear extract of HSG cells with anti-human CBP antibody showed a specific 270 kDa band, indicating the expression of CBP in HSG cells. The immunocytochemical analysis confirmed the nuclear localization of CBP. The transfection of HSG cells with a luciferase reporter plasmid harboring an RA-response element at the 5' upstream region of the reporter gene increased RA-dependent luciferase activity approximately 3-fold. Co-transfection with a CBP-expression plasmid and the luciferase reporter gene enhanced the RA-dependent transcription activation approximately 10-fold. The immunoprecipitates obtained with anti-CBP antibody exhibited a histone acetyl-transferase (HAT) activity 2-fold higher than that obtained with the control antibody, whereas the HAT activity of the immunoprecipitates with anti-SRC-1 and anti-p/CIP, which were used as comparisons, were only a little increased. The RA treatment had no effect on the level of HAT activity except in the case of using the immunoprecipitate obtained with anti-RARalpha, in which case it increased the activity. These findings indicate that CBP expressed in HSG cells mediates the RA-inducible growth and differentiation-regulating transcription activation in concert with the retinoic acid receptors. PMID- 14535630 TI - Comparing dual phase Tl-201 thyroid scan and fine-needle aspiration cytology to detect follicular carcinoma. AB - It remains difficult to pre-operatively differentiate follicular carcinoma and adenoma even under intraoperative investigations including histopathologic diagnosis of frozen sections. METHODS: We retrospectively compared dual phase thallium (Tl-201) thyroid scan and fine-needle aspiration (FNA) cytology to detect follicular carcinoma in 12 patients with cold thyroid nodules. Fine-needle aspiration cytology results were assessed and categorized by experienced pathologists into benign, suspicious, and malignant. Dual phase Tl-201 thyroid scan including an early (10-min) image and a delayed (3-hour) image were acquired after 2 mCi (74 MBq) of Tl-201 was injected intravenously. Dual phase Tl-201 thyroid scan findings were visually interpreted as positive and negative results. RESULTS: Based on FNA cytologic interpretation criteria, three nodules were diagnosed as benign, five nodules were diagnosed as suspicious, and four nodules were diagnosed as malignant. However, based on dual phase Tl-201 thyroid scan interpretation criteria, all of the 12 nodules could accurately diagnose as malignant. CONCLUSIONS: We conclude that dual phase Tl-201 thyroid scan was more accurate in diagnosing follicular carcinoma when comparing with FNA cytology. PMID- 14535631 TI - Serum ferritin levels in poorly- and well-controlled diabetes mellitus. AB - Serum ferritin level is related to body iron stores and is influenced by several diseases. We aimed to evaluate the association between serum ferritin concentration and the complications and nature of diabetes mellitus (DM). We examined association of ferritin concentration, fasting and post-prandial glucose levels and glycated hemoglobin in 329 patients with type 2 diabetes mellitus and 269 healthy controls. In the present study, 41 of 150 poorly controlled diabetic patients and 31 of 119 cases had hyperferritinemia. We confirmed that serum ferritin was increased in diabetic patients as long as glycemic control is not achieved. There was also a correlation between ferritin level and diabetic retinopathy. In conclusion, this study showed an independent positive association between serum ferritin concentration and markers of glucose homeostasis. PMID- 14535632 TI - Lipid profile and lipoprotein (a) as a risk factor for cardiovascular disease in women with subclinical hypothyroidism. AB - This study evaluated changes of lipids and lipoprotein (a) [Lp (a)] as atherosclerotic risk factors and the effects of levothyroxine (LT4) treatment on these parameters in patient with subclinical hypothyroidism (SH), defined by increased concentrations of thyrotropin (TSH) and normal thyroid hormone concentrations. We prospectively included 35 female patients with SH and 30 healthy controls. Serum lipid measurements and clinical score as well as fT3, fT4, and TSH levels were assessed at baseline. Body mass index (BMI) was also calculated. Women with proven SH received LT4 treatment that continued for 6 months. Twenty-six of 35 patients completed the study. At the end of treatment period, the same parameters were determined. Total cholesterol was significantly increased in patients with SH when compared with those of controls (p < 0.01), but increase of LDL-cholesterol (LDL-C) and Lp (a) were not significant (p > 0.05). In the levothyroxine-treated group, total cholesterol and LDL-C were significantly reduced when compared with the baseline values of women with SH (p < 0.05). Zulewski clinical score assessing symptoms and signs of hypothyroidism improved significantly with treatment (p < 0.001). In conclusion, serum lipids as important atherosclerotic risk factors increased before treatment and decreased with treatment. Levothyroxine therapy is effective in SH and improvements in serum lipids suggests that LT4 treatment also decreases the risk of atherosclerotic diseases. PMID- 14535633 TI - Correlation between bone mineral density and plasma lipids in Taiwan. AB - Many studies showed that depression is correlated with osteoporosis, while others showed that low cholesterol level is also related to depression. However, these relationships still remain controversial. Since the bone mineral density (BMD) is related to depression and depression is related to hypocholesterolemia, there might exist a correlation between BMD and plasma cholesterol levels. To prove this, we enrolled 5000 individuals, 2170 males, and 2830 females, who had health check-ups at a private clinic between 1998 and 1999. They were divided into three groups. Group 1 was composed of male subjects; Group 2, female subjects; and Group 3, females aged over 50 to exclude pre-menopausal females. Each subject had a routine physical examination, fasting blood drawing, BMD measured by dual energy x-ray absorptiometry (DEXA) over the wrist, and was given a questionnaire to answer. Between Groups 1 and 2, the females were significantly younger, had higher body mass index (BMI), total cholesterol (TC), high density lipoprotein cholesterol (HDLC), low density lipoprotein cholesterol (LDLC), and platelet, but lower BMD, fasting plasma glucose (FPG), triglycerides (TG), hemoglobin (Hgb), and white blood cell (WBC) count. As for Groups 1 and 3, all the aforementioned findings still remained the same except that the systolic blood pressure (SBP) was higher and diastolic blood pressure (DBP) was lower in Group 3. Our results showed that BMD is negatively related to age in males. In females, it is negatively correlated with age, FPG, PPG, SBP, DBP, TC, LDLC, TG, and Hgb, but positively related to BMI and platelet. However, for females in Group 3, BMD is only negatively related to age, FPG, SBP, and TG but positively related to BMI. Stepwise multiple regression analysis showed that the BMD is negatively related to age but positively related to BMI in both males and females. In Group 3, BMD is negatively related to age and FPG but positively related BMI. In conclusion, no correlation exists between BMD and cholesterol. This implies that the depression is not significantly related to cholesterol and/or BMD. This might be due to various confounding factors, which could affect their relationships. The negative correlation between BMD and FPG is only observed in females older than 50 years. Further studies are needed to clarify these relationships. PMID- 14535634 TI - The role of luteinizing hormone and prolactin in the regulation of insulin receptors in Leydig cells of the adult rat. AB - The role of luteinizing hormone (LH), prolactin and their combination in the regulation of insulin receptors in Leydig cells was studied. Leydig cells were isolated from adult male Wistar rats and measurement of insulin binding and internalization was done by incubating the cells with a saturating concentration of 125I-insulin in the presence or absence of different doses of unlabeled LH/insulin. LH exposure (100 and 200 ng dose) caused a significant increase in Leydig cell surface and internalized insulin receptor concentrations. Prolactin at all doses was ineffective in inducing a significant change in insulin receptor concentration. Under basal condition, Leydig cell surface binding of 125I-insulin was greater than the internalization at 34 degrees C but at 4 degrees C, surface binding remained lower than that at 34 degrees C with negligible internalization. Internalization of insulin receptors was measured by incubating the cells at 4 degrees C for 16h and then rapidly incubating at 34 degrees C for various time intervals (60, 120, and 180 min). LH/PRL or LH + PRL did not induce any significant change in the internalization of 125I-insulin at 60 and 120 min. The rate of internalization was greater at 120 min in basal as well as LH/PRL exposed Leydig cells, compared to 60 min of incubation. Prolactin alone did not evoke any appreciable change in internalization of 125I-insulin compared to basal at all three time points tested. Total and acid soluble release of 125I-insulin recorded a significant increase in Leydig cells exposed to LH, which was marginally potentiated when prolactin was added along with LH. Monensin treatment of Leydig cells prevented the recycling of insulin receptors to the cell surface and thereby suppressed the surface binding and enhanced the internalized 125I insulin. Under cycloheximide treatment, neither surface bound nor internalized 125I-insulin recorded a significant change compared to their respective basal values. It is concluded from the present study that LH has dose-dependent biphasic effects on insulin receptors in Leydig cells by modulating the internalization and intracellular processing of hormone-receptor complexes but prolactin has no such effects. PMID- 14535635 TI - Effects of estriol on the proliferation and differentiation of human osteoblastic MG-63 cells. AB - Estriol has been showed to prevent bone loss in osteoporotic rats and postmenopausal women, but the mechanisms remain unclear. In the present study, we evaluated the effect of estriol on osteoblastic MG-63 cells in vitro, and compared its action with 17beta-estradiol (E2). Cell proliferation was determined by measuring total cell numbers and [3H]thymidine incorporation. Cell function was studied by measuring alkaline phosphatase (ALP) activity and secreted osteocalcin. Our data showed that estriol stimulated MG-63 cells proliferation in a dose-dependent manner, but had no influence on ALP activity in MG-63 cells and osteocalcin production. Compared with estriol treatment, E2 showed a stronger proliferation. Estrogen receptor (ER) alpha and beta expression in MG-63 cells can be detected by Western immunoblot analysis, and the proliferative response to E2 and estriol can be all abrogated by ER antagonist ICI 182,780. In conclusion, estriol stimulates osteoblastic MG-63 cells proliferation, but has no effects on differentiation. The proliferative response to estriol is mediated by the ER. These results suggest that estriol has an effect on osteoblastic proliferation, and this may contribute to its actions on prevention of bone loss. PMID- 14535636 TI - 1alpha,25-dihydroxyvitamin D3 regulates the expression of membrane-type matrix metalloproteinase-1 in normal human osteoblast-like cells. AB - Recently, membrane-type matrix metalloproteinase-1 (MT1-MMP) was identified, and found that it can activate proMMP-2 on the cell membrane, degrade bone matrix, and participate in bone formation. Since bone is a target tissue of 1alpha,25 dihydroxyvitamin D3 [1alpha,25(OH)2D3], in the present study we observed the effects of 1alpha,25(OH)2D3 on MTI-MMP expression, and proMMP-2 activation in normal human ostcoblast-like cells (hOB). Western immunoblots showed 1alpha,25(OH)2D3 time and dose dependently stimulated MTI-MMP production. By ELISA, we found that the activation of proMMP-2 in cultures of hOB was intensified by 1alpha,25(OH)2D3. Our studies suggest that 1alpha,25(OH)2D3 induces MMP-2 activation in part by up-regulating MTI-MMP expression, and since MTI-MMP plays a role in bone metabolism, the induction of MT1-MMP levels by 1alpha,25(OH)2D3 in hOB cells may contribute to a new mechanism by which 1alpha,25(OH)2D3 promotes bone formation and stimulates bone resorption. PMID- 14535638 TI - Biotechnology for food, energy, and industrial products: new opportunities for bio-based products. PMID- 14535637 TI - Novel roles for the rho subfamily of GTP-binding proteins in succinate-induced insulin secretion from betaTC3 cells: further evidence in support of the succinate mechanism of insulin release. AB - We have previously demonstrated regulatory roles for Rho subfamily of G-proteins in glucose- and calcium-induced insulin secretion. Herein, we examined regulation by these proteins of insulin secretion from betaTC3 cells elicited by mitochondrial fuels, such as the succinic acid methyl ester (SAME). Preincubation of these cells with Clostridium difficile toxin-B (200 ng/mL), which monoglucosylates and inactivates Cdc42 and Rac1, markedly decreased (> 70%) SAME induced insulin secretion. Furthermore, exposure of betaTC3 cells to GGTI-2147 (20 microM), a selective inhibitor of the requisite prenylation of Rac1 and Cdc42, significantly reduced (> 80%) SAME-induced insulin release, suggesting that post-translational prenylation of these proteins is necessary for SAME induced insulin release. Western blot analysis indicated localization of Cdc42, Rac1, and Ras in the beta cell mitochondrial fraction. Confocal microscopy revealed a modest, but inconsistent, increase in the association of either Rac1 or Cdc42 with Mitotracker, a mitochondrial marker, following exposure to SAME. These data suggest that activation of preexisting intramitochondrial Rac1 and Cdc42 may be sufficient to regulate SAME-induced insulin secretion. Together, our findings support a role for G-proteins in insulin secretion at a step dependent on mitochondrial metabolism. They also identify mevalonate-derived, isoprenoid modified Rho G-proteins as specific signaling molecules in recently proposed succinate mechanism of insulin release. PMID- 14535639 TI - Effect of expandable clays and cometabolism on PAH biodegradability. AB - Pyrene and phenanthrene degradation was examined in both single and binary slurry systems for three different natural soils. It was found that the amount of total expandable clays (smectite and vermiculite) was in a good agreement with the achieved rate and extent of biodegradation. For instance, the intrinsic phenanthrene biodegradation rate was 626 microg/L/day for the soil with the largest expandable clay and 3203 microg/L/day for the soil with the least. Similarly, the smallest total pyrene biodegradation (65%) was found for the soil rich in expandable clays, compared to an 82% pyrene reduction in the soil that had the lowest amount. Mass transfer limitation after compound sorption to the clays was more pronounced for the more hydrophobic pyrene. In the presence of phenanthrene, total pyrene biodegradation increased by 2 to 7% due to cometabolism, while the total phenanthrene biodegradation was only enhanced by 0.5 to 5% in the binary system. This research demonstrated that expandable clays might govern the substrate availability to microorganisms and microbial accessibility to substrates. Therefore, the contribution of organic matter and expandable clays to sorption, desorption and biodegradation should be taken equally into account in order to better understand complex bioremediation issues. PMID- 14535640 TI - Toxic effects of orimulsion on rainbow trout Oncorhynchus mykiss. AB - GOAL, SCOPE AND BACKGROUND: Orimulsion (stable emulsion of natural bitumen and water) is a new imported industrial fuel in Lithuania. No data on its toxicity to fish is freely available. The aim of this study was to investigate sensitivity of rainbow trout (Oncorhynchus mykiss) to acute and chronic toxicity of orimulsion and to estimate the Maximum Acceptable Toxicant Concentration (MATC) of orimulsion to fish. METHODS: Laboratory tests were conducted on rainbow trout in all stages of development (embryos, larvae, adults). Acute toxicity (96-hour duration) and long-term (28 or 60-day duration) tests evaluating the wide range spectrum of biological indices were performed under semi-static conditions. RESULTS AND DISCUSSION: Median lethal concentration (96-hour LC50) values and their 95% confidence intervals derived from the tests were: 0.1 (0.09-0.12) to embryos, 0.06 (0.05-0.07) to larvae and 2.22 (2.02-2.43) to adult fish, and 28 day LC50 to adult fish was found to be 0.26 (0.21-0.32) g/l of total orimulsion respectively. The acute toxicity of orimulsion to rainbow trout can be characterised by a narrow zone of toxic effect and a sharp boundary between lethal and sublethal concentrations. The lowest 'safe' or 'no-effect' concentration values of total orimulsion obtained in long-term tests were equal to 0.09 g/l to adult fish, 0.019 g/l to embryos, and 0.0017 g/l to larvae. Proposed value of 'application factor' for orimulsion was found to be equal to 0.03. Since orimulsion has the property to disperse in all water volume, its toxic effect on fish can be characterised by the combined effects of dispersion and water-soluble-fraction. CONCLUSIONS: Maximum Acceptable Toxicant Concentration (MATC) of 0.0017 g/l of total orimulsion to fish was derived from long-term tests based on the most sensitive parameter of rainbow trout larvae (relative mass increase at the end of the test). According to substance toxicity classification accepted for Lithuanian inland waters, orimulsion can be referred to substances of 'moderate' toxicity to fish. RECOMMENDATIONS AND OUTLOOK: For prediction and evaluation of toxic impact of orimulsion accident spills on fish, some recommendations should be given. Since orimulsion has the property to disperse in all water volume during short time periods, the amounts of both spilled orimulsion and polluted water should be ascertained. Once both parameters are known, the real concentration of orimulsion in the water body must be determined. Then this concentration must be compared with 'safe' concentration to fish. By use of 'application factor' 0.03, approximate MATC for other fish species can be estimated when only acute toxicity data (96-hour LC50 value) is available. PMID- 14535641 TI - The changes in common carp blood after short-term zinc exposure. AB - Blood zinc level, hematological parameters and blood cell morphology were evaluated in common carp immediately after 3 h exposure to 20 mg dm(-3) of zinc (Zn0), and in 24, 48 and 96 hours after the end of it (Zn24, Zn48, Zn96). Blood zinc level in the non-exposed fish was 8 mg dm(-3), reached a maximum of 20 mg dm(-3) in Zn48, while it dropped to 9 mg dm(-3) in Zn96. Zinc caused a stress reaction in fish indicated by an increase in hematocrit value in Zn0, and elevated plasma glucose level and trombocytosis which persisted until the end of the experiment. Zinc-exposed fish showed an increased frequency of abnormal erythrocytes, and a compensatory release of immature erythrocytes to the blood stream. In zinc-treated fish, leukocyte count initially increased and subsequently decreased significantly below the control level due to a drop in lymphocyte number. Lymphocyte viability was reduced, and abnormal lymphocytes appeared. A decreased count of juvenile neutrophiles, and reduced phagocyte activity also occurred. The results indicate possible zinc-induced disturbances in both specific and non-specific immune mechanisms. PMID- 14535642 TI - Environmental impacts of coastal aquaculture in eastern Mediterranean bays: the case of Astakos Gulf, Greece. AB - GOAL, SCOPE AND BACKGROUND: Over the last decade Greece has become a leading country in the EU as concerns the cage farming of seabream and seabass. A strong debate has risen, however, about the environmental impacts of aquacultures in the coastal areas. The present paper deals with this problem and it is based on measurements of physico-chemical parameters in the water column, particulate matter and sediments in the area of Astakos Gulf, a coastal embayment in western Greece where three big fish farms are currently operating. METHODS: Water samples were collected by using Hydro-Bios sampling bottles, whereas a prototype sediment trap was installed under a fish cage for the collection of particulate matter. Temperature, dissolved oxygen, pH and salinity were measured in situ using portable equipment. Nutrients were determined by standard spectrophotometric methods. Trace metals were determined by Atomic Absorption Spectrometry. Dissolved organic carbon was determined by a Shimadzu 5000A carbon analyzer, whereas organic carbon in sediments was determined titrimetrically. RESULTS AND DISCUSSION: No clear eutrophication incidents have been identified, although the water column near the fish farms was enriched in nutrients and organic carbon. A sludge 'blanket' covers considerable parts of the seabed and is enriched in colloidal organic carbon and trace metals (Cd, Cu, Fe and Zn) that come from unused fish food. The biodegradation of this sludge leads to the development of anoxic conditions followed by the formation of undesirable gases, precipitation or remobilization of metals and the extinction of benthic fauna. CONCLUSION: The operation of fish farms at the coastal area of Astakos Gulf, and probably in similar Mediterranean gulfs, affects the marine environment, particularly in the vicinity of the cages. The most significant influence concerns the near-bottom water layer. The environmental impacts depend on the amount of food given to fishes, the mode of feeding, the fish density in cages, the annual production and the years of unit operation. The hydrology and the geomorphology of the area are also critical factors for its environmental quality. RECOMMENDATION AND OUTLOOK: The success of the fish-farming sector in the Mediterranean is accompanied by environmental and, in some extents, by social and marketing problems. These problems, derived from the rapid development of fish farming, can be solved only through an integrated management, using methods such as environmental impact assessment, risk assessment, economic evaluation, vulnerability assessments, resource accounting, cost-benefit analysis and outcome-based monitoring. PMID- 14535643 TI - Effects of a fire-retardant chemical to fathead minnows in experimental streams. AB - BACKGROUND: Each year millions of liters of fire-retardant chemicals are applied to wildfires across the nation. Recent laboratory studies with long-term fire retardant chemicals indicate a significant photoenhanced toxicity of products containing sodium ferrocyanide corrosion inhibitors. Our objective of this study was to determine the toxicity of fire-retardant chemicals to fathead minnows during exposure in experimental outdoor streams. METHODS: Stream tests were conducted to determine the potential toxicity of a pulse of exposure as might occur when fire retardant chemical is rinsed from the watershed by rainfall. Two artificial 55-meter experimental streams were dosed with different concentrations of Fire-Trol GTS-R, or uncontaminated for a control. Replicate groups of fathead minnows were added to screened containers (10 fish per container) and exposed to retardant chemicals in the recirculating flow of the stream for up to 6 hours. RESULTS AND DISCUSSION: Under field conditions toxicity of GTS-R only occurred in the presence of sunlight. When GTS-R was tested on sunny days, 100% mortality occurred. However, when tested during heavily overcast conditions, no mortality occurred. CONCLUSIONS: Lethal concentrations of cyanide were measured when GTS-R with YPS exposures were conducted under sunny conditions, but not under cloudy conditions, indicating that a minimum UV level is necessary to induce toxicity as well as the release of cyanide from YPS. The toxicity observed with GTS-R was likely associated with lethal concentrations of cyanide. Rainwater runoff following applications of this fire-retardant at the recommended rate could result in lethal concentrations in small ponds and streams receiving limited water flow under sunny conditions. RECOMMENDATIONS AND OUTLOOK: In addition to avoiding application to aquatic habitats, it is important to consider characteristics of the treated site including soil binding affinity and erosive properties. PMID- 14535644 TI - Generic scenario for the impact of cooling lubricants into the hydrosphere. AB - According to directive 93/67/EEC of the European Commission, this paper deals with the generic exposition evaluation of the environmental concentration of cooling Lubricant chemicals from the metal working industry into the hydrosphere. After considering the relevant life-cycle steps and the selection of a representative point source for the 'reasonable worst case', the emission per day Elocal(water) is determined. It leads to the predicted environmental concentration (PEClocal(water)) for the local stage in the compartment water. In order to gain the PEClocal(water) for an example--a corrosion inhibitor as additive in a cooling lubricant--, the relevant emission paths and the corresponding representative point source are described for the reasonable worst case. For non-water-miscible cooling lubricants, none of the operations in the life cycle leads to a release into the compartment water. To evaluate the hazard potential for cooling lubricant chemicals, the complete risk assessment has to be done. Also, the assessment has to be done for all high production volume chemicals, new substances and existing hazard chemicals. This means that even industrial categories like chemicals used in the textile industry or biocides and others have to be evaluated. PMID- 14535645 TI - Biodegradation of anthracene in the roots and growth substrate of poplar cuttings. AB - Following their exposure to anthracene, the roots of Populus nigra L. Loenen showed traces of 9 substances classed as products of biodegradation. The main substances detected were phthalic acid and 9,10-anthraquinone, followed by hydroxyanthracene and methoxyanthracene and five other compounds which could not be identified. Due to the relatively low concentration of degradation products found in the roots, further degradation to lower molecular compounds are discussed. The presence of 9,10-anthraquinone as the main product of the degradation of anthracene was also evident in the control tests with unplanted sandy substrate, although the content was higher in the planted series of tests. As a non-sterile approach was chosen, it may be assumed that a microbial degradation for 9,10-anthraquinone took place in the control series. However, it is difficult to differentiate clearly between a microbial degradation of anthracene in the substrate and metabolization in the roots due in part to the absence of specific degradation products in the various reaction areas. PMID- 14535646 TI - Investigation of the complexation of metal-ions by strong ligands in fresh and marine water. AB - SCOPE: The detection and investigation of metal ions bound in strong complexes in natural waters is a difficult task, due to low concentration of the metal ions themselves, and also of the strong ligands, which, moreover, are often not of a well-defined composition. Here, a method is proposed for the investigation of the speciation of metal ions in natural waters. OBJECTIVE AND METHOD: It is based on the sorption of metal ions on strongly sorbing ion exchange resins, i.e. complexing resins. For this reason the method is called Resin Titration. It has been shown in previous investigations that the concentration of metal ion totally sorbed by a particular resin, and its reaction coefficient in the solution phase in the presence of the resin, can be determined from the sorption data using a simple relationship. Here, a data treatment (the Ruzic linearization method) is proposed for also determining the concentration of the ligands responsible for the complex in equilibrium with the resin. RESULTS: The method was applied to data obtained by Resin Titration of a freshwater and a seawater. Copper(II) and aluminium(III) were considered, using Chelex 100 as a titrant, due to its strong sorbing properties towards these metal ions. The results were: the total metal concentration in equilibrium with the resin, the side reaction coefficients, and the concentration of ligands. In all these cases the ligands forming very strong complexes were found to be at concentration lower than that of the metals. CONCLUSION: The Ruzic linearization method allows the determination of the concentration of the ligands forming very strong complexes in equilibrium with Chelex 100. The reaction coefficient was better determined by the calculation method previously proposed for RT. The ligands responsible for the strong complexes were found to be at low concentration, often lower than that of the metal ions considered. The metal in the original sample is partly bound to these ligands, since the complexes are very strong. Only a part of the metal is linked to weaker ligands, or free. PMID- 14535648 TI - A new route for the chemical valorisation of lactose. AB - A totally protected di-O-benzyl derivative of triacetonlactose dimethyl acetal was transformed into a 4'-hexeno disaccharide by elimination of acetone with t BuOK in DMF and subsequently in 5'-C-methoxy derivative by oxidation with MCPBA in methanol as a solvent. The hydrolysis of this latter compound affords 2,6-di-O benzyl-L-arabino-aldohexosos-5-ulose, which by intramolecular aldol condensation with DBU gives an inosose that was stereoselectively reduced to epi-inositol. Therefore our synthetic strategy offers a new and simple method to transform lactose into carbocyclic monosaccharide analogues. PMID- 14535647 TI - Aromatic photonitration in homogeneous and heterogeneous aqueous systems. AB - This work describes the nitration of aromatics upon near-UV photolysis of nitrate and nitrite in aqueous solution and upon photocatalytic oxidation of nitrite in TiO2 suspensions. Phenol is used in this work as a model aromatic molecule and as a probe for *NO2/N2O4. The photoinduced nitration of phenol in aqueous systems occurs upon the reaction between phenol and *NO2 or N2O4, and is enhanced by the photocatalytic oxidation of nitrite to *NO2 by TiO2. Aromatic photonitration in the liquid phase can play a relevant role in the formation of nitroaromatics in natural waters and atmospheric hydrometeors, thus being a potential pathway for the condensed-phase nitration of aromatics. Furthermore, the photoinduced oxidation of nitrite to nitrogen dioxide suggests a completely new role for nitrite in natural waters and atmospheric aerosols. PMID- 14535649 TI - Bromine in waste incineration: partitioning and influence on metal volatilisation. AB - INTENTION, GOAL, SCOPE, BACKGROUND: The halogen bromine is far less abundant than chlorine, but it can be found at high concentrations in special materials like flame retarded plastics. The fate and effects of Br in waste incineration are not well understood. It may have similar implications like Cl for the volatilisation of heavy metals and the formation of low volatile organic compounds. Due to its lower oxidation potential, there is a risk of formation of elementary Br2 in the offgas. OBJECTIVE: Co-combustion tests of different types of Br containing plastic waste materials (up to 22%) and MSW in the TAMARA pilot plant for waste incineration were conducted to investigate the Br partitioning and the influence of Br on metal volatilisation. METHODS: The Br inventory of the fuel mix was elevated to approx. 1 wt-%. All input and output mass flows of the furnace have been sampled and the partitioning of Cl, Br, S, and a number of heavy metals, has been calculated on the basis of closed mass balances. RESULTS AND DISCUSSION: Organically-bound Br was typically released to more than 90% into the raw gas. Elementary Br2 was detected at high Br levels. Its presence was always analysed when all SO2 in the raw gas was oxidised to SO3. Br enhances the volatilisation of metals like K, Zn, Cd, Sn, Sb, and Pb out of the fuel bed principally in the same way as Cl. The tests gave strong indication that the promoting influence of the halogens on metal volatilisation is more pronounced than that of the fuel bed temperature. The volatilised metals are condensated on the fly ashes and are discharged along with the filter ashes. CONCLUSIONS: As long as a surplus of SO2 is present in the raw gas no Br2 is formed. Although the halogen induced transfer out of the fuel bed causes high concentrations of volatile metals in the filter ashes, a recovery is not economically feasible for the time being. The volatilisation gives no rise to metal emission problems as long as efficient dedusting is achieved. RECOMMENDATION AND OUTLOOK: If there is a risk of Br2 formation, in wet scrubbing a reducing agent has to be added to the neutral scrubber for efficient abatement. Filter ashes should be disposed of in a way that enables access for recovery in the future. The exact volatilisation characteristics of the various metals have to be studied in future using specifically tailored experiments. PMID- 14535650 TI - Heavy metals in plants and phytoremediation. AB - GOAL, SCOPE AND BACKGROUND: In some cases, soil, water and food are heavily polluted by heavy metals in China. To use plants to remediate heavy metal pollution would be an effective technique in pollution control. The accumulation of heavy metals in plants and the role of plants in removing pollutants should be understood in order to implement phytoremediation, which makes use of plants to extract, transfer and stabilize heavy metals from soil and water. METHODS: The information has been compiled from Chinese publications stemming mostly from the last decade, to show the research results on heavy metals in plants and the role of plants in controlling heavy metal pollution, and to provide a general outlook of phytoremediation in China. Related references from scientific journals and university journals are searched and summarized in sections concerning the accumulation of heavy metals in plants, plants for heavy metal purification and phytoremediation techniques. RESULTS AND DISCUSSION: Plants can take up heavy metals by their roots, or even via their stems and leaves, and accumulate them in their organs. Plants take up elements selectively. Accumulation and distribution of heavy metals in the plant depends on the plant species, element species, chemical and bioavailiability, redox, pH, cation exchange capacity, dissolved oxygen, temperature and secretion of roots. Plants are employed in the decontamination of heavy metals from polluted water and have demonstrated high performances in treating mineral tailing water and industrial effluents. The purification capacity of heavy metals by plants are affected by several factors, such as the concentration of the heavy metals, species of elements, plant species, exposure duration, temperature and pH. CONCLUSIONS: Phytoremediation, which makes use of vegetation to remove, detoxify, or stabilize persistent pollutants, is a green and environmentally-friendly tool for cleaning polluted soil and water. The advantage of high biomass productive and easy disposal makes plants most useful to remediate heavy metals on site. RECOMMENDATIONS AND OUTLOOK: Based on knowledge of the heavy metal accumulation in plants, it is possible to select those species of crops and pasturage herbs, which accumulate fewer heavy metals, for food cultivation and fodder for animals; and to select those hyperaccumulation species for extracting heavy metals from soil and water. Studies on the mechanisms and application of hyperaccumulation are necessary in China for developing phytoremediation. PMID- 14535651 TI - Workshop 'ecotoxicology: scientific profile and practical needs'. PMID- 14535652 TI - The adenine nucleotide translocator: a new potential chemotherapeutic target. AB - Identification of new targets is of utmost importance for the development of efficient apoptosis-modulating drugs. This has become possible from the unraveling of the basic apoptosis mechanisms and notably, from the demonstration of the mitochondrial membrane permeabilization as a central rate-limiting step of numerous models of cell death. Indeed, molecular and pharmacological studies revealed that the adenine nucleotide translocator (ANT) could be a therapeutic target. First, ANT is a bi-functional protein. It mediates the exchange of cytosolic ADP and mitochondrial ATP, and contributes to apoptosis via its capacity to become a lethal pore. Second, both ANT functions are under the control of the (anti)-oncogenes from the Bax/Bcl-2 family, and third, agents as diverse as proteins, lipids, ions, pro-oxidants or chemotherapeutic agents directly modulate the pore-forming activity of ANT. Here, we will review the mode of apoptosis induction by various classes of chemotherapeutic agents, which all influence directly ANT pro-apoptotic function. Hopefully, this will yield several clues to the modulation of apoptosis from a therapeutic perspective. PMID- 14535653 TI - Tumor escape from immune response: mechanisms and targets of activity. AB - Immune system plays an important role in control of tumor progression. Effective antitumor immune response depends on close interaction of several elements of immune system. They include antigen-presenting cells, different subsets of T cells, B cells and NK cells. However, tumor cells developed a number of mechanisms to escape recognition and elimination by immune system. In this review we will discuss these mechanisms and address possible approaches to correct them. PMID- 14535654 TI - Thirty years of polyamine-related approaches to cancer therapy. Retrospect and prospect. Part 1. Selective enzyme inhibitors. AB - As soon as the natural polyamines (PAs), putrescine (Put), spermidine (Spd) and spermine (Spm), were recognized as ubiquitous constituents of eukaryotic cells, their involvement in growth-related processes attracted particular interest. The high activities of ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (AdoMetDC) in rapidly growing tissues and cells, particularly in tumour cells, suggested PA biosynthesis as a target for antineoplastic therapy. In the course of the years selective inhibitors have been developed for literally all enzymes of PA metabolism. Some became important as tools in the elucidation of the PA metabolic system, but only few of them were efficient as inhibitors of tumour growth. A major reason for the inefficacy of selective enzyme inhibitors as anticancer drugs is the sophistication of the system, which regulates intracellular PA pools. Selective blockade of a single enzyme induces changes of metabolism and transport, which compensate for the deficit. The selective impairment of tumour growth is in addition hampered by the ubiquitous occurrence of the PAs, their importance in normal functions of nearly all mammalian cells, and the ability or the mammalian organism to utilize exogenous (gastrointestinal) PAs. Among the inhibitors of PA-related enzymes, the ODC inactivator (R, S)-2 (difluoromethyl)ornithine (DFMO) became most famous. Although it was disappointing in most therapeutic attempts to use it as single drug, it has- based on its low toxicity--considerable potential in cancer chemoprevention, and it turned out to be a highly efficient anti-trypanosome agent. Very likely DFMO is suitable to improve the efficacy of some of the current cytotoxic drugs, and it may allow one to create new therapies in combination with other PA-directed drugs. Some of the less selective enzyme inhibitors, particularly those, which inhibit two or more enzymes of PA metabolism, appear to have had a chance to become practically useful, but they have not been developed energetically. Disregarding DFMO, the AdoMetDC inhibitor SAM486A is the only compound for which clinical trials were published. The future of this drug is unclear at present; presumably phase III clinical trials have been discontinued. One of the lessons that had to be learned from the work on selective enzyme inhibitors was that PA metabolism is a much more difficult target, than has been expected on the basis of the simplicity of the PA structures, and the simple reactions involved in their biosynthesis. In order to inhibit tumour growth several reactions or regulatory functions of PA metabolism have to be impaired at the same time. Recent efforts devoted to the development new types of anticancer drugs, which are based on the perturbation of PA metabolism by structural analogues of the natural PAs, take this message into account. These approaches are the topic of the 2nd part of this overview. PMID- 14535655 TI - Thirty years of polyamine-related approaches to cancer therapy. Retrospect and prospect. Part 2. Structural analogues and derivatives. AB - Owing to their role in growth-related processes the natural polyamines (PAs), putrescine (Put), spermidine (Spd) and spermine (Spm) were identified about 30 years ago as potential targets for the development of anticancer drugs. It was presumed that inhibition of a key enzyme of PA biosynthesis, followed by the depletion of the intracellular PA pools results in the prevention of cell growth. Initial efforts were nearly exclusively focused on the design and synthesis of selective inhibitors of the PA biosynthetic enzymes. This period is reviewed in the 1st part. Selective inhibition of ODC caused in various cell lines growth inhibition, but was usually not sufficient to inhibit tumour growth, because the PA regulatory system outbalances selective enzyme blockade by enhancing compensatory reactions, and because exogenous PAs are used if de nova synthesis is impaired. When these facts were recognized, new targets were envisaged. Among these the PA uptake system and the deregulation of PA homeostasis became most attractive. They are the major topic of the present 2nd part. Inhibition of PA uptake from the cellular environment is expected to improve the efficacy of drugs, which rely on the depletion of intracellular PA pools. During the past few years several potent inhibitors of the PA uptake system became known. However, more work will be needed to allow their assessment as anticancer drugs in combination with DFMO and other compounds capable of depleting PA pools. The PA transport system also offers the possibility to improve the accumulation by tumors of compounds, which are tethered to PA structures. This can be achieved for the following reasons: (a) Structural requirements of the PA uptake systems are not stringent. (b) Tumour cells accumulate PAs more avidly than most non transformed cells. (c) The transport rate for PAs is higher in cells with depleted PA pools, than in cells with a normal PA content. (d) In cells, which proliferate rapidly, PA depletion by biosynthesis inhibitors is more effective, than in slowly growing cells. The most actively pursued approach is currently based on the inhibition of tumour growth by cytotoxic structural analogues of the natural PAs. Some of these compounds mimic regulatory properties of the natural PAs. If a PA mimetic accumulates in cells, it induces catabolic processes, suppresses biosynthetic reactions, and depletes the pools of Put. Spd and Spm. N1,N11 -bis(ethyl)norspermine is a prototype of the PA mimetics. During the last decade a very large number of PA derivatives and structural analogues have been prepared, which are capable of inhibiting cell growth at low microM concentrations. Among the new PA-like structures several compounds were identified, which prevent cells from growing, without depleting PA pools to an extent that would be necessary to prevent cell growth. They may be considered as PA antagonists, although their mode of action is not well understood. A therapeutically useful drug has not yet been identified among the PA analogues. In many instances investigations were stopped at a preliminary stage. Recently synthesized compounds have not yet been pursued far enough to justify the initiation of clinical trials. Only very few toxicological results of the new structures have been reported, although the knowledge of the toxicology of Spm analogues is of eminent importance. PAs are ubiquitous cell constituents and are indispensable for normal cell function. However. extracellular PAs, and particularly extracellular Spm is cytotoxic and neurotoxic. These properties are shared by close structural analogues. A major difficulty in the development of PA analogues to therapeutically useful drugs is, therefore, the identification of structures, which do not share neurotoxic properties with Spm. Several tetramines are presently in early phases of clinical trials. It will be a matter of a few more years to allow one to decide, whether PA-related approaches of cancer therapy are a success or a failure. PMID- 14535656 TI - Melanocortins and their receptors and antagonists. AB - The melanocortins are a group of small protein hormones derived by post translational cleavage of the proopiomelanocortin (POMC) gene product. The known melanocortin hormones include alpha-melanocyte stimulating hormone (MSH), beta MSH, gamma-MSH and adrenocorticotropic hormone (ACTH). Five melanocortin receptors (MCIR through to MC5R) have been identified and most of these show tissue-specific expression patterns, as well as different binding affinities for each of the melanocortin hormones. The central melanocortin system consists of alpha-MSH, agouti-related protein (AGRP), MC3R and MC4R. AGRP and alpha-MSH are believed to be the natural antagonist and agonist respectively of MC3R and MC4R. This central melanocortin system is thought to play a fundamental role in the control of feeding and body weight. Knock-out mice models and genetic studies have pointed to the importance of the melanocortins in complex human pathways such as pigmentation, lipolysis, food intake, thermogenesis, sexual behaviour, memory and inflammatory response. Recently the melanocortins and their receptors have been the target for drug-based treatment of human physiological processes. MC3R and MC4R are likely targets for controlling body weight; MCIR may be used in the treatment of inflammation and MC2R for the treatment of glucocortical deficiency. A role for MCSR still remains unclear, but the evidence suggests an exocrine gland function. PMID- 14535657 TI - Nonmetric subadult skeletal sexing traits: I. A blind test of the accuracy of eight previously proposed methods using prehistoric known-sex mummies from northern Chile. AB - While a number of nonmetric sex-related traits have been proposed, the accurate assignment of sex to subadult skeletal materials is generally considered problematic. Eight previously proposed nonmetric traits of the ilia and mandible have been demonstrated by other researchers to be related to the known-sex of subadults, yet there has been relatively little research attempting to evaluate the utility of these traits using independent collections of known-sex subadult skeletal remains. These eight traits include: elevation of the auricular surface, angle of the greater sciatic notch, depth of the greater sciatic notch, the "arch criteria," curvature of the iliac crest, gonial eversion, mandibular protrusion, and mandibular arcade shape. The present study blindly tested these eight nonmetric traits using 85 autopsied prehistoric known-sex mummified subadult remains from northern Chile that range in age between newborn and 15 skeletal years of age. The two primary questions examined for each trait are: (1) are the different character states for each of the eight traits related to known-sex, and (2) which of the eight traits are accurate enough for use in forensic cases? These questions are examined for each trait by both sex and age class so as to uncover potential sex- and age-related strengths and weaknesses for each of the characteristics. The results indicate that all eight of the previously proposed subadult sex-related traits are indeed related to known-sex, but that there is a great deal of variation by both sex and age in terms of the strength of congruent sex-related associations. With the exception of gonial eversion, all of the traits produced statistically significant chi2 values for their associations with known-sex when all subadult remains were considered. However, when all subadults are considered, only four traits demonstrated acceptable levels of accuracy for forensic applications. These traits include the arch criteria (82.3%), angle of the sciatic notch (80.7%), depth of the sciatic notch (79.0%), and mandibular arcade shape (77.6%). For subadults ranging in age from newborn to five skeletal years of age, only depth of the sciatic notch (81.5%) and the arch criteria (81.5%) approach acceptable levels of accuracy for use in forensic cases. The implications of these results are discussed and recommendations for bioarchaeological and forensic applications are made. PMID- 14535658 TI - Development of an Alu-based, real-time PCR method for quantitation of human DNA in forensic samples. AB - Determining the amount of human DNA extracted from a crime scene sample is an important step in DNA profiling. The forensic community relies almost entirely upon a technique (slot blot) to quantitate human DNA that is imprecise, time consuming, and labor intensive. We have previously described a method for quantitation of human DNA based on PCR amplification of a repetitive Alu sequence that uses a fluorescence plate reader. This manuscript describes and validates a variation of this assay using real-time PCR and SYBR Green I for quantitation. The advantages of the real-time assay over the plate reader assay are: reduced hands-on time, lower assay cost, and a greater dynamic range. The main disadvantage is the cost of the real-time instrument. However, for those forensic laboratories with access to a real-time instrument, this Alu-based assay has a dynamic range of 16 ng to 1 pg, is sensitive, specific, fast, quantitative, and uses only 2 microL of sample. PMID- 14535659 TI - A potential metallographic technique for the investigation of pipe bombings. AB - This study was conducted in an attempt to develop a metallographic method for the investigation of pipe bombings. Three common pipe materials, ASTM A53 steel, AISI 304L stainless steel, and 6061-T6 aluminum, were shock-loaded using five high explosives and three propellants. The explosives used were ANFO, Composition C4, C6 detasheet, nitroglycerine-based dynamite, and flake TNT. The propellants used were FFFFg black powder. Red Dot smokeless powder, and Turbo Fuel A. The post blast microstructure, hardness, and, in the case of 304L, transformed martensite content were examined for each test. The damage done to the microstructure was found to increase with increasing detonation velocity of the explosives and increase in pressure generated by the shock-metal interaction. Material hardness and, in the case of 304L, martensite content showed a sharp increase followed by a plateau as the shock pressure and detonation velocity increased. PMID- 14535660 TI - Vacuum collection of gunpowder residues from clothing worn by shooting suspects, and their analysis by GC/TEA, IMS, and GC/MS. AB - Experiments were conducted to collect gunpowder (propellant) residues from shooters' clothing by vacuum and to analyze them by gas chromatography/thermal energy analyzer (GC/TEA), ion mobility spectrometry (IMS), and gas chromatography/mass spectrometry (GC/MS). The residues were collected on fiberglass and Teflon filters using the portable vacuum sampler, all supplied with the IMS instrument. Several solvents were examined for the extraction of the propellant components from the filters. The extracts were centrifuged and/or filtered, concentrated by evaporation, and analyzed without any additional clean up procedure. Based on the results of the study, an operational method for analysis of gunpowder residues was introduced into casework without changing the present operational technique for gunshot (primer) residue (GSR) analysis on clothing implemented by the Israel Police. In the modified method, the clothing is first sampled by double-side adhesive-coated aluminum stubs (the tape-lift method) for GSR analysis (the existing method), followed by vacuum collection for propellant residue examination. The issue of interpretation of the analytical results is discussed. PMID- 14535661 TI - Fingerprint pattern restoration by digital image processing techniques. AB - Fingerprint evidence plays an important role in solving criminal problems. However, defective (lacking information needed for completeness) or contaminated (undesirable information included) fingerprint patterns make identifying and recognizing processes difficult. Unfortunately. this is the usual case. In the recognizing process (enhancement of patterns, or elimination of "false alarms" so that a fingerprint pattern can be searched in the Automated Fingerprint Identification System (AFIS)), chemical and physical techniques have been proposed to improve pattern legibility. In the identifying process, a fingerprint examiner can enhance contaminated (but not defective) fingerprint patterns under guidelines provided by the Scientific Working Group on Friction Ridge Analysis, Study and Technology (SWGFAST), the Scientific Working Group on Imaging Technology (SWGIT), and an AFIS working group within the National Institute of Justice. Recently, the image processing techniques have been successfully applied in forensic science. For example, we have applied image enhancement methods to improve the legibility of digital images such as fingerprints and vehicle plate numbers. In this paper, we propose a novel digital image restoration technique based on the AM (amplitude modulation)-FM (frequency modulation) reaction diffusion method to restore defective or contaminated fingerprint patterns. This method shows its potential application to fingerprint pattern enhancement in the recognizing process (but not for the identifying process). Synthetic and real images are used to show the capability of the proposed method. The results of enhancing fingerprint patterns by the manual process and our method are evaluated and compared. PMID- 14535662 TI - Development and validation of the Y-PLEX 5, a Y-chromosome STR genotyping system, for forensic casework. AB - A genotyping system, Y-PLEX 5, has been developed for use in human identification. The Y-PLEX 5 enables simultaneous amplification of five polymorphic short tandem repeat (STR) loci residing on the Y-chromosome, which are DYS389I, DYS389II, DYS439, DYS438, and DYS392. As little as 0.1 ng of template DNA can be used for analysis. The specificity of the amplification reaction enabled analysis of male DNA in a male: female DNA mixture at a ratio of 1:600. Mean stutter values ranged from 3.60-10.97%. Among the different primates investigated, the DNA from orangutan exhibited amplification at DYS438 locus and from gorilla at DYS439 and DYS438 loci. The DNA from cat, dog, and horse did not yield any amplified product. Studies on development of the genotyping system, generation and description of the allelic ladder, and validation of the multiplex PCR as per the FBI Director's Quality Assurance Standards were carried out. Y-STR allele and haplotype frequencies in two populations were generated. The data indicate that the Y-PLEX 5 genotyping system is sensitive and reliable, and can be used in human forensic and male lineage identification cases. PMID- 14535663 TI - STR DNA typing: increased sensitivity and efficient sample consumption using reduced PCR reaction volumes. AB - Improvements in detection limits/sensitivity and lower sample consumption are potential benefits of reducing PCR reaction volumes used in forensic DNA typing of crime scene samples. This premise was studied first with experimental mixtures and a nine-loci megaplex, which demonstrated stochiometric amplification and accurate detection. Next, adjudicated casework samples were subjected to amplification under 15 different template DNA to PCR reaction volume ratios. Reduction of PCR reaction volume and DNA down to 10 microL and 0.500 ng, respectively, produced identical profiles with the same signal intensity and heterozygous allele peak height ratio (HR). Reduction to 5 microL and 0.063 ng yielded HR values that were slightly affected in one to three STR loci. PCR reaction volume reduction can enhance detection and sensitivity while reducing the consumption of irreplaceable crime scene samples. PMID- 14535665 TI - Analysis of human fecal material for autosomal and Y chromosome STRs. AB - Human stool samples from eight volunteers were stored under various conditions and extracted by three different procedures. Fecal material and tissue paper soiled with fecal material obtained from a crime scene were also extracted. Extracted DNA was amplified using the AmpFlSTR Profiler Plus, AmpFlSTR COfiler, and the AmpFlSTR Identifiler PCR amplification kits for the detection of the autosomal STR allelic patterns. DNA extracted from the male volunteers and from the soiled tissue paper evidence sample was also amplified using the Y-PLEX 6 and Y-PLEX 5 amplification kits. Analysis of the amplified products was carried out by capillary electrophoresis on the ABI PRISM 310 Genetic Analyzer. Autosomal and Y-STR profiles obtained from the fecal material were concordant with the results from the donors' buccal swabs. PMID- 14535664 TI - AmpFlSTR profiler Plus short tandem repeat DNA analysis of casework samples, mixture samples, and nonhuman DNA samples amplified under reduced PCR volume conditions (25 microL). AB - As part of the validation of the AmpFlSTR Profiler Plus short tandem repeat (STR) system, under reduced polymerase chain reaction (PCR) volume conditions (i.e., 25 microL), a total of 275 casework samples were processed. Examples of profiles are presented along with amplification conditions to improve the odds of obtaining balanced and complete profiles for samples showing partial results or profiles with a descending slope. Data collected and used to develop our interpretation guidelines are included. From the mixture studies, full profiles were obtained for minor contributors, using 2 ng of DNA, with ratios of 10:1 or 1:20 and using 1 ng of DNA, with ratios of 10:1 and 1:8. The specificity of the Profiler Plus amplification reaction performed in 25 microL was examined and confirmed using a large spectrum of nonhuman DNAs. This report supports the use of the AmpFlSTR Profiler Plus STR system for casework DNA typing under reduced PCR volume conditions. PMID- 14535666 TI - Developmental validation of a real-time quantitative PCR assay for automated quantification of human DNA. AB - Our laboratory has developed an automated real-time quantitative PCR assay for detecting human DNA. The assay utilizes an in-house, custom-designed TaqMan-MGB sequence-specific probe (CFS-HumRT) and the ABD 7900HT SDS platform. Developmental validation has followed TWGDAM (1) guidelines and demonstrates that the assay is primate specific, is highly sensitive, yields consistent results, and works with human DNA extracted from a variety of body fluid stains. When operating within the dynamic range of the system using high-quality DNA samples. the technique yields similar quantification results to our current QuantiBlot assay with the added benefit of time saving through automation. Furthermore, the QPCR assay identifies how much amplifiable DNA is in a sample and thus has the potential to predict PCR success in downstream applications such as STR analysis. PMID- 14535667 TI - Forensic applications of chemical imaging: latent fingerprint detection using visible absorption and luminescence. AB - Chemical imaging technology is a rapid examination technique that combines molecular spectroscopy and digital imaging, providing information on morphology, composition, structure, and concentration of a material. Among many other applications, chemical imaging offers an array of novel analytical testing methods, which limits sample preparation and provides high-quality imaging data essential in the detection of latent fingerprints. Luminescence chemical imaging and visible absorbance chemical imaging have been successfully applied to ninhydrin, DFO, cyanoacrylate, and luminescent dye-treated latent fingerprints, demonstrating the potential of this technology to aid forensic investigations. In addition, visible absorption chemical imaging has been applied successfully to visualize untreated latent fingerprints. PMID- 14535668 TI - The development of reduced size STR amplicons as tools for analysis of degraded DNA. AB - New multiplex PCR sets of commonly used short tandem repeat (STR) markers have been developed to produce PCR products that are reduced in size when compared to standard commercial STR kits. The reduction in size of these amplicons can facilitate the examination and analysis of degraded DNA evidence by improving amplification efficiency. This "miniSTR" approach will permit current forensic practitioners to use STR markers and instrumentation already present in their laboratories and to generate genotyping data that is directly comparable to reference samples and searchable through the FBI's Combined DNA Index System (CODIS) databases. This paper discusses the development of these new primer sets and presents some initial results in the analysis of degraded and aged DNA samples. A method for removal of problematic fluorescent dye artifacts is also described. Comparison studies in over 100 samples have verified that these miniSTR primers can provide fully concordant results to commercial STR kits and can provide improved signal from degraded DNA specimens. These miniplex sets should prove valuable in the analysis of samples where allele dropout and reduced sensitivity of larger STR alleles occurs. PMID- 14535669 TI - Characterization of condom lubricant components using Raman spectroscopy and Raman chemical imaging. AB - A condom can be described as a protective sheath used as a contraceptive or to protect against sexually transmitted diseases. However, individuals also use condoms during the commission of sexual assaults to prevent identification through deposited biological material. Raman spectroscopy offers a novel approach to identifying the presence of condom lubricant components. Furthermore, Raman chemical imaging expands on conventional Raman spectroscopy to characterize multiple condom lubricant components simultaneously in a manner that effectively demonstrates heterogeneous sample mixtures both spectrally and spatially. Known reference materials, liquid and solid lubricant components of common condom brands were successfully characterized using Raman dispersive spectroscopy and Raman chemical imaging without extensive sample preparation inherent to other analytical methods. The characterization of these materials demonstrates the potential of this technique to become a routine screening method for condom lubricants. This preliminary investigation provides a basis for future studies to determine the feasibility of Raman spectroscopy and Raman chemical imaging for condom lubricant trace detection in case type samples. PMID- 14535670 TI - Optimization of the separation of organic explosives by capillary electrophoresis with artificial neural networks. AB - The separation of 12 explosives by capillary electrophoresis was optimized with the aid of artificial neural networks (ANNs). The selectivity of the separation was manipulated by varying the concentration of sodium dodecyl sulfate (SDS) and the pH of the electrolyte, while maintaining the buffer concentration at 10 mM borate. The concentration of SDS and the electrolyte pH were used as input variables and the mobility of the explosives were used as output variables for the ANN. In total, eight experiments were performed based on a factorial design to train a variety of artificial neural network architectures. A further three experiments were required to train ANN architectures to adequately model the experimental space. A product resolution response surface was constructed based on the predicted mobilities of the best performing ANN. This response surface pointed to two optima; pH 9.0-9.1 and 60-65 mM SDS, and pH 8.4-8.6 and 50-60 mM SDS. Separation of all 12 explosives was achieved at the second optimum. The separation was further improved by changing the capillary to an extended cell detection window and reducing the diameter of the capillary from 75 microm to 50 microm. This provided a more efficient separation without compromising detection sensitivity. PMID- 14535671 TI - Filtration based DNA preparation for sexual assault cases. AB - Police departments in the United States currently have as many as 500,000 unprocessed swabs taken from rape victims. The standard method for purifying sperm from these swabs is to resuspend first all cells and to digest selectively the excess of the victim's epithelial cells. The intact sperm are then separated from the contaminating solubilized DNA by centrifugation, careful removal of supernatant, and extensive washing of the sperm pellet, all steps that are difficult to automate. Vacuum driven filtration is an alternative method for separating sperm from digested epithelial cells that requires only pipetting steps and can be readily automated in a 96 well format. Sperm DNA is enriched 45 fold using this process and the yield of PCR ready DNA is roughly 20% of the amount originally present on the swab. PMID- 14535672 TI - A novel multiplex PCR system consisting of Y-STRs DYS441, DYS442, DYS443, DYS444, and DYS445. AB - We have developed a new sensitive multiplex PCR system consisting of five male specific and polymorphic tetranucleotide STRs--DYS441 (GDB: 10013873), DYS442 (GDB: 10030304), DYS443 (GDB: 10807127), DYS444 (GDB: 10807128), and DYS445 (GDB: 10807129) on the Y chromosome. Fifty pg DNA per 10 microL reaction volume was required for the correct typing of five STRs. Using this system, the five Y-STRs were correctly typed from blood and semen stains that had been stored for several years at room temperature. PMID- 14535673 TI - Evidence in support of self-declaration as a sampling method for the formation of sub-population DNA databases. AB - Well constructed sub-population databases are fundamental to the application of DNA-based forensic statistics. The size of such databases can affect the ability to examine adequately statistical or population genetic features, and the integrity of both the DNA profile and associated ethnicity information is also of importance. Use of short tandem repeat (STR) DNA profiling technology and the thoughtful construction of the governing legislation has seen large databases of DNA profiles collated for the four major sub-populations of New Zealand. Examination of the data illustrates the suitability of self-declaration as a means of categorizing samples on the basis of ethnicity. PMID- 14535674 TI - Modifications to the silver physical developer. AB - The silver physical developer is currently the most successful reagent used for visualizing the water-insoluble components (e.g., lipids) of latent prints on porous surfaces. It is normally used after the amino acid visualizing reagents (e.g., ninhydrin and DFO) are used. This work found that the performance of the current formulation of silver physical developer is strongly reduced when the water used is changed from the usual distilled water to the more purified reverse osmosis/deionized (RO/DI) water. Based on numerous experiments involving the systematic variation of the component concentrations, the performance was restored and even improved by reducing the concentration of all the components (except that of the ferric salt) and by including malic acid in the formulation. These modifications resulted in a new silver physical developer formulation that performs as well as or better than the current formulation and is less expensive to make. PMID- 14535675 TI - Elemental analysis of glass fragments by ICP-MS as evidence of association: analysis of a case. AB - Inductively coupled plasma mass spectrometry (ICP-MS) was used to analyze glass evidence from a case in which a person broke a window in each of 15 vehicles in a parking lot in order to gain entry into the vehicles. The results of the analysis by traditional methods, which measure the properties of color, thickness, density, and refractive index, are also reported. A total of 15 known samples representing the windows on the cars and 42 questioned glass fragments recovered from the suspect and the police vehicle where the suspect was sitting were submitted for analysis. Density comparisons separated one of the known samples into three samples, increasing the number of known samples from 15 to 17. The concentrations of 16 elements were measured for all but three of the samples using an external calibration ICP-MS method with internal standardization. Color assessment (non-instrumental) separated the 17 known samples into two groups, and refractive index measurements resulted in six groups when the Emmons double variation method was used and ten groups when the Glass Refractive Index Measurement 2 (GRIM2) system was used. Elemental analysis, by itself, differentiated all of the known samples from each other and associated four of the known sample fragments with several of the questioned sample fragments. The informing power of RI, density, and elemental analysis comparisons is evaluated and a summary of the case results is reported. PMID- 14535676 TI - Identification of DYS385 allele variants by using shorter amplicons and Northern Thai haplotype data. AB - Primers currently used for amplification of locus DYS385 on the Y chromosome give quite long amplicons that are difficult to resolve on native polyacrylamide gels. We therefore designed new primers that give much shorter products for easy separation. With this system, two allele variants that were characterized by sequencing as 14.1 (GAAA --> GAAAA) and 14.2 (GA insertion), were identified in two unrelated Northern Thai men due to their decreased mobility. Interestingly, initial capillary electrophoresis mistyped the 14.1 variant amplified with the traditional primers as allele 14. Forty different haplotypes were found in 147 Northern Thais with a haplotype diversity of 0.9430. Haplotype 13-18 (frequency 0.136) is the most frequent one: for comparison: haplotype 14-18 (frequency 0.074) was found to be the most common in a population from Bangkok (n = 95) in an earlier study. This striking difference within Thailand's population probably results from genetic differences in the founding populations. PMID- 14535677 TI - Suicide by fire in a car trunk: a case with potential pitfalls. AB - We present an unusual case in which an adult female committed suicide by fire while enclosed in the trunk of her vehicle. A finding of suicide was reached only because several key investigative facts were uncovered. Without these discoveries the case may still remain open as a suspicious death. Fire is rarely used as a mechanism of suicide in Western culture, and, as such, similar cases should be reported to advance our recognition of this phenomenon. PMID- 14535678 TI - A web-based survey of odontologist's opinions concerning bitemark analyses. AB - Within the field of forensic dentistry, the detection, collection, and analysis of bitemarks remains one of the most contentious areas. Attempts at the production of consensus documents have produced documents such as the ABFO's Guidelines for Bitemark Evidence Collection. Despite this, the range of differing analysis techniques, allied with a varied opinion base on the robustness of bitemark conclusions has led to polarized views within the profession. The purpose of this study was to survey forensic dentists to obtain their views on a number of crucial components of bitemark theory and contentious areas within the discipline. Using a web-based survey, 14 questions were asked of respondents. Seventy-two odontologists completed the survey, with 38% being of Diplomate status, 10% had completed 20 or more bitemark cases, and 20% between 10 and 20 cases, 91% of respondents believed that the human dentition was unique, with 78% believing that this uniqueness could be represented on human skin during the biting process. Seventy percent believed that they could positively identify an individual from a bitemark, and 22% stated that the statistical tool, the product rule, should be applied to bitemark conclusions. Over half of the odontologists used overlays for bitemark analysis. with a digital method of production the most popular. The implications of these and other findings are discussed. PMID- 14535679 TI - Dentists' qualifications affect the accuracy of radiographic identification. AB - This study assessed the effects of forensic odontologists' training and experience upon the accuracy of their dental radiographic identifications. Forty participating odontologists with various levels of training and experience completed a Web-based survey of their qualifications and then completed nine Web based radiographic identification cases. They made their identifications using the American Board of Forensic Odontology Categories and Terminology for Body Identification. The results indicate that odontologists with high levels of training and experience performed significantly more accurately than those with lower levels. We conclude that high levels of training and experience in forensic odontology should be developed, maintained, and required of dentists who participate in a forensic team dealing with challenging identification cases. PMID- 14535680 TI - The use of dental aging techniques in forensic odontological practice. AB - Many studies have been published describing numerous techniques to determine the aging of individuals from dental tissues. However, few case reports demonstrate how such techniques can be employed by the forensic odontologist undertaking casework. Indeed, many of the techniques are highly complex and utilize equipment not generally available to odontologists outside of the university or hospital systems. This paper describes five cases where dental aging was used for identification in the absence of materials for any other techniques. In each case the Bang and Ramm method was employed using sectioned teeth. Teeth were sectioned, photographed, and scaled in image analysis software. Bang and Ramm equations were employed, and in each case an age was derived. The estimated age was supplied to the coroner, who was able to use the information to positively identify five individuals for whom no other identification system was feasible. PMID- 14535681 TI - Insect faunal succession on decaying rabbit carcasses in Punjab, India. AB - Insect faunal succession on decaying rabbit carcasses was carried out at Punjabi University, Patiala (Punjab), India, from March 1997 to December 1999. Four stages of decomposition were recognized, i.e., fresh, bloated, decay, and dry. A total of 38 insect species belonging to four orders and 13 families were recorded. Diptera, Coleoptera, and Hymenoptera dominated the carrion fauna. Calliphorids were the first to arrive in all the seasons of the year. Five species of Calliphoridae, four of Sarcophagidae, ten of Muscidae, and one each from Anthomyiidae and Otitidae were observed on rabbit carcasses. Representatives of six Coleopteran families, i.e., Staphylinidae, Histeridae, Cleridae, Dermestidae, Tenebrionidae, and Silphidae, were recorded. Eight species belonging to family Formicidae (Hymenoptera) were also collected during the present studies. Only one species of Lepidoptera was observed on carrion. PMID- 14535682 TI - Mortality in cerebral palsy (CP): the importance of the cause of CP on the manner of death. AB - Cerebral palsy (CP) is a nonprogressive motor deficit present or sustained in the perinatal period. We reviewed the files of the Office of Chief Medical Examiner of the City of New York for the 1997-2001 interval seeking those with any mention of cerebral palsy. There were 26 such cases, including 18 natural deaths, three accidents, two homicides, two therapeutic complications, and one death classified as undetermined. Proper reporting and careful investigation of these deaths is required for accurate certification of cause and manner of death, as well as for adequate tracking of these deaths for public health purposes. PMID- 14535683 TI - Inflicted compressional asphyxia of a child. AB - Compressional or traumatic asphyxia is a well recognized entity to most forensic pathologists. The vast majority of reported cases have been accidental. The case reported here describes the apparent inflicted compressional asphyxia of a small child. A review of mechanisms and related controversy regarding proposed mechanisms is discussed. PMID- 14535684 TI - Suicidal hangings in jail using telephone cords. AB - In a three month period of 2000, three suicide hanging deaths involving the use of telephone cords occurred in Texas jails and were autopsied at the Southwestern Institute of Forensic Sciences in Dallas. In each case, the telephone was located within the cell, allowing the prisoners unhindered access at any time. Because of these incidents, two of the jails shortened their receiver cords to a total length of 6-8 in. The telephones were otherwise unaltered, and are still in the same locations. The third jail replaced their entire phone with a cordless telephone. Despite the attention that these cases received, standards have not been set regarding the type or placement of telephones used within jail cells, and a fourth incident in 2002 underscores this ongoing problem. These three incidents highlight the need to provide telephones that, if placed within holding cells or other jail cells, do not provide a possible means of suicide. PMID- 14535685 TI - Central nervous system consequences of an unusual body disposal strategy: case report and brief experimental investigation. AB - The body of a 73-year-old man was dismembered by his female companion for the purpose of covert disposal. The method employed included skillful separation of body parts with hacksaw and knife, piecemeal disposal of fragments, and prolonged boiling of the decapitated head. The latter treatment resulted in marked shrinkage of cranial dura mater, separation of dura mater from skull, and extrusion of brain fragments into the resultant enlarged epidural space through a dural defect due to the disproportionately greater shrinkage of dura mater compared to brain parenchyma. This resulted in curd-like brain fragments filling an enlarged epidural space and overlying a shrunken, leathery dura mater. The cranial dura mater, still adherent to the skull base, resembled a "shrunken brain" in contour but contained only the remnants of brain tissue not already extruded through the dural defect. This unusual thermal artifact is rarely illustrated or mentioned in forensic literature. The development of this postmortem artifact likely requires the presence of a specific combination of conditions which must be, but apparently rarely are, simultaneously present. PMID- 14535686 TI - Parasomnia pseudo-suicide. AB - Complex behaviors arising from the sleep period may result in violent or injurious consequences, even death. Those resulting in death may be erroneously deemed suicides. A series of case examples and review of the pertinent literature are provided to increase awareness of the possibility that some apparent "suicides" are the unfortunate, but unintentional, consequence of sleep-related complex behaviors and therefore are without premeditation, conscious awareness, or personal responsibility. The correct cause-of-death determination in such cases may have profound social, religious, and insurance implications for surviving friends and family members. PMID- 14535688 TI - The direct detection and identification of staining dyes from security inks in the presence of other colorants, on currency and fabrics, by laser desorption mass spectrometry. AB - Laser desorption mass spectrometry (LDMS) is useful for the direct desorption and MS analysis of dyes off materials such as paper. Here it is shown that staining dyes, produced by currency degradation devices, such as those used by financial institutions or armored transport companies, can be detected by LDMS. These staining dye packs are commonly used in bank security programs to deter theft or unauthorized removal of currency from teller stations or from ATM cash cassettes. Working automatically, these technologies release a security dye to degrade the surface of the notes and possibly mark the assailant involved in the attack. These dyes can be characterized and identified directly from paper currency, and from fabrics, and uniquely identified in the presence of other dyes that are used in the printing and dying of such materials, by LDMS. In these experiments, no extraction step is required. A pulsed UV laser directly irradiates a paper or fabric sample--colorants are desorbed, ionized and detected using time-of-flight MS. Results shown here suggest that dyes such as Basic Red 1:1 and Basic Violet 11:1 are used in such devices. PMID- 14535687 TI - The forensic analysis of thermal transfer printing. AB - Thermal transfer printing refers to printing processes that utilize heat to produce an image by either physical or chemical means or by a combination of both. As the technology has improved and the supplies have become less expensive, the use of thermal printing in the personal and business markets has increased significantly. Specifically, dye diffusion thermal transfer and thermal mass transfer have become predominant in the production of counterfeit credit cards, drivers' licenses, and other types of documents produced on plastic media. Chemical analysis by means of thin layer chromatography (TLC) has proven to be useful in characterizing various types of inks (e.g., writing and inkjet inks). In this study, the authors examined 81 different samples that included a total of 54 printer samples (43 photographic prints on paper and eleven plastic card samples) and 27 printer ribbons. A new TLC method was developed and tested utilizing a solvent system (80% n-hexane, 3% methyl ethyl ketone, and 17% ethyl acetate) that is capable of producing excellent resolution. PMID- 14535689 TI - Two fatal cases involving concurrent use of methamphetamine and morphine. AB - We report on two cases of simultaneous administration of methamphetamine (MAP) and morphine (MOR) with hyperthermia. The blood levels of MAP and MOR were toxic and putatively lethal, respectively, although hyperthermia is a known cause of intoxication due to MAP rather than MOR. In Japan, MAP is the most predominant cause of drug intoxication. The presented cases suggest that MOR may exert synergistic effect on hyperthermia due to the MAP intoxication, together with experimental findings. PMID- 14535690 TI - A Spanish population study of the STR loci D2S1338, D19S433, Penta D, and Penta E. PMID- 14535691 TI - Population data of the STRs D2S1338 and D19S433 from northeast of Spain. PMID- 14535692 TI - Chinese population data on DXS6797 and GATA144D04 loci. PMID- 14535693 TI - Polymorphism of two STR loci on chromosome 21 in a Chinese population. PMID- 14535694 TI - Allele frequencies of two Y-STRs in a Chinese population. PMID- 14535695 TI - Distribution of allele frequencies of one VNTR and two STR loci in five population groups of South India. PMID- 14535696 TI - Microsatellite markers in the population from Rio Grande do Norte, Northeastern Brazil. PMID- 14535697 TI - Haplotype frequency distribution of two autosomal microsatellites (D12S66 and D12S67) among five endogamous population groups in India. PMID- 14535698 TI - DNA polymorphism study at D1S80, DYS19, DYS287, and DYF155S2 in two tribal populations from Central India. PMID- 14535699 TI - Forensic assessment of ACTBP2 (SE33) microsatellite. PMID- 14535700 TI - Population genetics for the CODIS core STR loci in the population of Northeastern Poland. PMID- 14535701 TI - Gene frequencies for two Y-chromosomal STR loci in Chinese population. PMID- 14535702 TI - Commentary on: Thompson WC, Taroni F, Aitken CGG. How the probability of a false positive affects the value of DNA evidence. J Forensic Sci 2003;48(1):47-54. PMID- 14535703 TI - Commentary on: Thompson WC, Taroni F, Aitken CGG. How the probability of a false positive affects the value of DNA evidence. J Forensic Sci 2003 Jan.;48(1):47-54. PMID- 14535704 TI - Letter to editor-autopsies on executed federal prisoners. PMID- 14535705 TI - Commentary on: Dettmeyer R, Kandolf R, Baasner A, Banaschak S, Eis-hubinger AM, Madea B. Fatal parvovirus B19 myocarditis in an 8-year-old boy. J Forensic Sci 2003;48(1): 183-186. PMID- 14535706 TI - Semantic differences between "tumescent liposuction," "tumescent anesthesia," and "tumescent technique". PMID- 14535707 TI - Design and synthesis of C3-symmetric Lewis(x) antigen. AB - [structure: see text] C(3)-Symmetric glycoconjugates carrying three equivalent Lewis(X) antigens or beta-lactosides were synthesized from p-nitrophenyl glycosides and trimesic acid via regio- and stereocontrolled glycosylation reactions. An (1)H NMR study has shown that the C(3)-symmetric glycoconjugates soluble in water provide useful probes to investigate the Ca(2+)-dependent Lewis(X)-Lewis(X) association. PMID- 14535708 TI - Regiodependent luminescence quenching of biotinylated N-sulfonyl-acridinium-9 carboxamides by avidin. AB - [reaction: see text] Biotin was conjugated to chemiluminescent N sulfonylacridinium-9-carboxamides at the N-10 or 9-position carboxamide. Upon binding to avidin, the light output of the N-10 derivative (8) was quenched up to 92% upon triggering with basic peroxide, while the 9-position carboxamide conjugate (9) was quenched only 33%. The utility of this effect was demonstrated in a model homogeneous chemiluminescence assay. PMID- 14535709 TI - Dynamic multivalent lactosides displayed on cyclodextrin beads dangling from polymer strings. AB - [reaction: see text] Lactose-appended cyclodextrin derivatives have been synthesized and threaded onto hydrophobic polymers in aqueous solution to form dynamic multivalent lactosides for binding to lectins. The threading process, which proceeds quickly, can be observed by one- and two-dimensional NMR spectroscopies. PMID- 14535710 TI - Phase-vanishing methodology for efficient bromination, alkylation, epoxidation, and oxidation reactions of organic substrates. AB - [reaction: see text] In cases where both reactants in a phase-vanishing reaction are less dense than the fluorous phase, an alternative to the U-tube method is to employ a solvent with greater density than the fluorous phase, such as 1,2 dibromoethane. This modification has been successfully applied to the methylation of a phenol derivative with dimethyl sulfate and to the m-CPBA-induced epoxidation of alkenes, N-oxide formation from nitrogen-containing compounds, and S-oxide or sulfone formation from organic sulfides. PMID- 14535711 TI - Chemoselective nucleophilic attack on N-acyl derivatives of (S)-ethyl 4,4 dimethyl pyroglutamate (DMPG). AB - [reaction: see text] Heteronucleophiles and C-nucleophiles chemoselectively react with N-acyl (S)-ethyl 4,4-dimethyl pyroglutamate (DMPG) affording esters, amides, and ketones in high yield. The intramolecular process allows the stereoselective formation of beta-hydroxy acids likely by formation and ring opening of the corresponding beta-lactones PMID- 14535712 TI - Structurally novel Bi- and tricyclic beta-lactams via [2 + 2] cycloaddition or radical reactions in 2-azetidinone-tethered enallenes and allenynes. AB - [reaction: see text] Thermolysis of beta-lactam-tethered enallenyl alcohols gave tricyclic ring structures via a formal [2 + 2] cycloaddition of the alkene with the distal bond of the allene, while the tin-promoted radical cyclization in 2 azetidinone-tethered allenynes proceeded to provide bicyclic beta-lactams containing a medium-sized ring. The access to cyclization precursors was achieved by regio- and stereoselective metal-mediated carbonyl allenylation of 4 oxoazetidine-2-carbaldehydes in an aqueous environment. PMID- 14535713 TI - N,N-dimethyl glycine-promoted Ullmann coupling reaction of phenols and aryl halides. AB - [reaction: see text] Ullmann-type diaryl ether synthesis can be performed at 90 degrees C using either aryl iodides or aryl bromides as the substrates under the assistance of N,N-dimethylglycine. PMID- 14535714 TI - Palladium-catalyzed cross-coupling of acetates of Baylis-Hillman adducts and potassium organotrifluoroborates. AB - [reaction: see text] The cross-coupling of potassium organotrifluoroborates and acetates of Baylis-Hillman adducts proceeds readily in moderate to excellent yield in the presence of Pd(OAc)(2). The reaction tolerates hindered trifluoroborate salts, and the process is stereoselective. PMID- 14535715 TI - Efficient solid-phase synthesis of symmetric norbinaltorphimine derivatives. AB - [reaction: see text] We describe an efficient solid-phase synthesis of symmetric norbinaltorphimine (norBNI) derivatives 2. Pyrrole formation involving the homocoupling of two solid-supported ketones 6, followed by chemoselective and sequential N-alkylation, provided N-substituted norBNI derivatives 2. Use of this methodology led to the combinatorial synthesis of 120 norBNI derivatives. PMID- 14535716 TI - Oxa-di-pi-methane photochemical rearrangement of quinuclidinones. Synthesis of pyrrolizidinones. AB - [reaction: see text] The oxa-di-pi-methane (ODPM) photochemical rearrangement, a triplet-sensitized sigmatropic 1,2-acyl shift of beta,gamma-enones, was successful utilizing methyl and heptyl 1-aza-3-carboalkoxybicyclo[2.2.2]oct-2-en 5-ones (quinuclidinones) as the photoprecursors. The cyclopropane of the heptyl ester tricyclic photoproduct could be opened with lithium dimethylcuprate or via hydrogenolysis to produce the corresponding pyrrolizidinone skeletons. PMID- 14535717 TI - Palladium-catalyzed Heck reaction on 1-alkoxy-1,3-dienes: a regioselective gamma arylation of alpha,beta-unsaturated carbonyl compounds. AB - [reaction: see text] alpha,beta-Unsaturated acetals afford, in the presence of the LIC-KOR superbase, 1-alkoxybuta-1,3-dienes. These substrates cross couple with aryl derivatives in the presence of Pd catalyst (Heck conditions) in a regio and stereoselective mode. With dialkyl acetals, the reaction affords arylated dienes; on the other hand, in the case of 1,3-dioxane derivatives, the final outcome of the process formally corresponds to the direct gamma-arylation reaction of the starting alpha,beta-unsaturated material. PMID- 14535718 TI - Ethylene-promoted intermolecular enyne metathesis. AB - [reaction: see text] Intermolecular enyne metathesis between functional group rich alkynes and vinyl ethers was promoted by ethylene cometathesis. The concentration of ethylene was optimized to suppress the competing formation of butadiene through background ethylene metathesis. The role of ethylene appears to be both protective and rate enhancing. PMID- 14535719 TI - Highly stable keto-enamine salicylideneanilines. AB - [reaction: see text] Highly stable NH salicylideneanilines have been prepared by reaction of 1,3,5-triformylphloroglucinol with aniline derivatives. The NH form was confirmed by X-ray crystallographic data, as well as by NMR studies. A convenient one-step synthesis of triformylphloroglucinol is also reported. PMID- 14535720 TI - Efficient sialylation with phenyltrifluoroacetimidates as leaving groups. AB - [reaction: see text] Sialylation with N-phenyltrifluoroacetimidates as leaving groups and a catalytic amount of TMSOTf as promoter compares favorably with the previous protocols for direct sialylation and expand in essence the scope of the Schmidt glycosylation reaction. PMID- 14535721 TI - Synthesis of new chiral monodentate phosphite ligands and their use in catalytic asymmetric hydrogenation. AB - [reaction: see text] New monodentate phosphite ligands have been developed from axially chiral biphenols, which show excellent enantioselectivity in the Rh(I) catalyzed hydrogenation of dimethyl itaconate. The new chiral ligand system is suitable to create libraries and possesses fine-tuning capability. PMID- 14535722 TI - Indium-mediated tandem radical addition-cyclization-trap reactions in aqueous media. AB - [reaction: see text] Tandem carbon-carbon bond-forming reactions were studied by using indium as a single-electron-transfer radical initiator. The radical addition-cyclization-trap reaction of a substrate having a vinyl sulfonamide group and an olefin moiety proceeded smoothly in aqueous media. The radical addition-cyclization reaction of hydrazone gave the functionalized cyclic products. PMID- 14535723 TI - Enantioselective total synthesis of (+)-azimine and (+)-carpaine. AB - [reaction: see text] The enantioselective total syntheses of (+)-azimine and (+) carpaine have been developed, starting with (S)-1,2,4-butanetriol as a single source of chirality. The key common feature in these syntheses involves stereoselective intramolecular hetero-Diels-Alder reaction of an acylnitroso compound. The critical macrocyclic dilactonization of the N-Cbz derivatives of azimic acid and carpamic acid was efficiently achieved by using the Yamguchi macrocyclization conditions. PMID- 14535724 TI - 2-Aryl and 2-heteroaryl indoles from 1-alkynes and o-iodotrifluoroacetanilide through a domino copper-catalyzed coupling-cyclization process. AB - [reaction: see text] A general method for the synthesis of 2-aryl and 2 heteroaryl indoles from aryl iodides and 1-alkynes through a domino copper catalyzed process is reported. The best results have been obtained with [Cu(phen)(PPh(3))(2)]NO(3) in the presence of K(3)PO(4) in toluene or dioxane at 110 degrees C. 2-Aryl and 2-heteroaryl indoles can also be isolated in good yields by using catalysts derived from CuI and PPh(3) in dioxane at 110 degrees C. PMID- 14535725 TI - A synthesis of (+)-salvadione-A. AB - [reaction: see text] The p-benzoquinone shown is converted to the novel hexacyclic triterpene salvadione-A in four steps. PMID- 14535726 TI - Unprecedented intermolecular transamidation reaction of N-carbamoylmethyl-N' tosylguanidines. AB - [reaction: see text] N-Carbamoylmethyl-N'-tosyl guanidine 2 reacts easily with primary alkylamines to afford substituted carboxamides 3. The reaction proceeds via a five-membered-ring intermediate 5, which could be isolated, and features a rare example of an intermolecular transamidation reaction under mild conditions. PMID- 14535727 TI - Asymmetric synthesis of functionalized trans-2,6-disubstituted piperidines with N sulfinyl delta-amino beta-ketoesters. Synthesis of (-)-lasubine I. AB - [reaction: see text] The hydroxy-directed reduction of 1,2-dehydropiperidines with the "ate" complex of DIBAL-H and n-BuLi affords functionalized trans-2,6 disubstituted piperidines. This methodology was employed in the asymmetric synthesis of the quinolizidine alkaloid (-)-lasubine I. PMID- 14535728 TI - Selective synthesis of the para-quinone region of geldanamycin. AB - [structure: see text] The quinone portion of the ansamycin geldanamycin was made with complete selectivity from the 1,4-dihydroquinone generated from a 1,4-bis methoxymethyl (MOM) ether intermediate. Palladium catalysis with air gave the desired product in 98% isolated yield. The structure was established using NMR, UV, and X-ray analysis with comparisons to geldanamycin, ortho-quino-geldanamycin and a model compound. PMID- 14535729 TI - Highly efficient asymmetric Lewis acid catalysis with platinum group complexes of conformationally flexible 1,3-butadiene-bridged diphosphines, NUPHOS. AB - [structure: see text] Palladium and platinum complexes of conformationally flexible 1,3-butadiene-bridged diphosphines NUPHOS can be resolved with (S)-BINOL at elevated temperatures to afford diastereopure delta-[(NUPHOS)M[(S)-BINOL]] (M = Pd, Pt). The homochiral Lewis acid complexes delta-[(NUPHOS)M][OTf](2), generated by protonation of delta-[(NUPHOS)M[(S)-BINOL]] with trifluoromethanesulfonic acid, catalyze the Diels-Alder reaction between acryloyl N-oxazolidinones and cyclopentadiene to give ee values up to 96%. The corresponding enantiopure dichlorides delta-[(NUPHOS)PtCl(2)] react with AgClO(4) to form highly efficient catalysts that give good endo/exo selectivities and high endo enantioselectivity. PMID- 14535730 TI - Aminoborohydrides 15. The first mild and efficient method for generating 2 (dialkylamino)-pyridines from 2-fluoropyridine. AB - [reaction: see text] Lithium aminoborohydride (LAB) reagents promote the amination of 2-fluoropyridine under mild reaction conditions, providing 2 (dialkylamino)pyridines in excellent yield and purity. Treatment of 2 fluoropyridine with 1.1 equiv of lithium aminoborohydride at room temperature affords complete conversion after 1 h. This is the first general way by which 2 (dialkylamino)pyridines may be directly obtained from fluoropyridines under ambient reaction conditions. 2-Chloropyridine can also be converted to 2 (dialkylamino)pyridine by simply increasing the number of LAB equivalents and the reaction temperature. PMID- 14535731 TI - The Pt-catalyzed carboselenation of alkynes by selenoesters. AB - [reaction: see text] The Pt-catalyzed carboselenation of terminal alkynes with selenoesters provided vinylselenides regio- and stereoselectively in moderate yields. PMID- 14535732 TI - Intramolecular cycloaddition reactions of silyl nitronate tethered to vinylsilyl group: 2-nitroalkanols as precursors for amino polyols. AB - [reaction: see text] A method for converting 2-nitroalkanols to precursors for stereodefined amino polyols is described. Diphenylvinylsilylation of the 2 nitroalkanols' hydroxy groups and subsequent silyl nitronate generation by using TMS-Cl and Et(3)N in CH(3)CN at 0 degrees C to room temperature led to fused bicyclic heterocycles through stereoselective intramolecular nitronate-olefin [3 + 2] cycloaddition reaction. Some examples for transforming the cycloadducts to amino polyols are also presented. PMID- 14535733 TI - Conversion of D-glucose to cyclitol with hydroxymethyl substituent via intramolecular silyl nitronate cycloaddition reaction: application to total synthesis of (+)-cyclophellitol. AB - [reaction: see text] A diastereoisomeric mixture of 1-nitro-6-heptene-2,3,4,5 tetraol derivative (A) was prepared by Henry reaction between d-glucose-based aldehyde and nitromethane. Only the (2S)-isomer of A led to cyclitol (B) via nitronate-olefin cycloaddition on treatment with TMS-Cl and Et(3)N in the presence of catalytic DMAP followed by acid treatment. (+)-Cyclophellitol (1) was synthesized from B in eight steps. PMID- 14535734 TI - Stereoselective construction of cis-2,6-disubstituted tetrahydropyrans via the reductive etherification of delta-trialkylsilyloxy substituted ketones: total synthesis of (-)-centrolobine. AB - [reaction: see text] The stereoselective intramolecular reductive etherification of delta-trialkylsilyloxy substituted ketones with catalytic bismuth tribromide and triethylsilane provides a convenient method for the construction of cis-2,6 disubstituted tetrahydropyrans. This method was highlighted in the key step of an expeditious total synthesis of the antibiotic, (-)-centrolobine. PMID- 14535735 TI - Stereocontrolled synthesis of psi[CH(CF3)NH]gly-peptides. AB - [reaction: see text] A novel class of peptidomimetics having a stereogenic [CH(CF(3))NH] replacement for a [CONH] peptide bond has been synthesized. The new compounds have been obtained in a stereocontrolled fashion using a kinetically controlled aza-Michael addition of chiral alpha-amino acid esters to trans-3,3,3 trifluoro-1-nitropropene. The stereoselectivity is strongly influenced by the solvent, the base, its stoichiometry, and the R side-chain. Diastereomeric ratios higher than 11:1 were achieved using H-Val-OtBu.HCl in toluene with 1.1 equiv of DIPEA. PMID- 14535736 TI - A synthesis of L-vancosamine derivatives from non-carbohydrate precursors by a short sequence based on the Marshall, McDonald, and Du Bois reactions. AB - [reaction: see text] The carbamate-protected l-vancosamine glycal, viewed as a universal precursor for vancosamine derivatives, was prepared by a short scheme based on diastereoselective addition of an allenyl stannane to a lactaldehyde ether, the tungsten-catalyzed alkynol cycloisomerization, and the rhodium catalyzed C-H insertion of a carbamate nitrogen. This sequence is a prototype for a new and efficient strategy for the synthesis of 3-amino sugar derivatives. The key intermediate was elaborated to the silyl ether of N,N-dimethyl vancosamine glycal. PMID- 14535737 TI - Novel trifunctional building blocks for fluorescent polymers. AB - [reaction: see text] Herein, we describe the synthesis of fluorescent 2 (arylsulfonyl)methacrylates and its polymers. These novel trifunctional monomers, possessing a fluorescent arylsulfonyl (ArSO(2)) group, an alkyl group (R), and a polymerizable olefin, serve as useful building blocks for functionalized fluorescent polymers. PMID- 14535738 TI - Lewis acid-mediated reactions of alkyl azides with alpha,beta-unsaturated ketones. AB - [reaction: see text] Alkyl azides react with saturated ketones upon treatment with Lewis acids to afford ring-expansion products through the azido-Schmidt reaction, but this reaction does not proceed when alpha,beta-unsaturated ketones are used. In this study, alkyl azides were reacted with enones in the presence of Lewis acids to give enaminones (vinylogous amides), which formally involve a ring contraction reaction. The mechanism and scope of this reaction is discussed. PMID- 14535739 TI - Total synthesis and absolute configuration of otteliones A and B, novel and potent antitumor agents from a freshwater plant. AB - [reaction: see text] The first enantioselective total synthesis of otteliones A and B, biologically important and structurally novel natural products, has been successfully achieved. This total synthesis fully confirms the absolute configuration of these natural products. PMID- 14535740 TI - Novel synthetic approach to 6,7-dihydro-5H-imidazo[1,5-a]-pyrazin-8-ones. AB - [reaction: see text] A novel route to highly substituted chiral 6,7-dihydro-5H imidazo[1,5-a]pyrazine-8-ones starting from Meldrum's acid is disclosed. The key features of the methodology are the incorporation of amino esters as a chiral pool and facile mild intramolecular cyclization to form the pyrazine ring. Incorporation of various substituents at different stages of the synthesis from various building block sets makes this methodology readily amenable to parallel synthesis. PMID- 14535741 TI - Selective fluorescence quenching of 2,3-diazabicyclo[2.2.2]oct-2-ene by nucleotides. AB - [reaction: see text] The fluorescence quenching of 2,3-diazabicyclo[2.2.2]oct-2 ene (DBO) by nucleotides has been studied. The quenching mechanism was analyzed on the basis of deuterium isotope effects, tendencies for exciplex formation, and the quenching efficiency in the presence of a molecular container (cucurbit[7]uril). Exciplex-induced quenching appears to prevail for adenosine, cytidine, and uridine, while hydrogen abstraction becomes competitive for thymidine and guanosine. Compared to other fluorescent probes, DBO responds very selectively to the type of nucleotide. PMID- 14535742 TI - Palladium(0)-mediated desymmetrization of meso tetraols: an approach to the C3 C17 bis-oxane segment of phorboxazoles A and B. AB - [reaction: see text] meso-Tetraol bis(allylic acetates) 2 and 5 were synthesized via two-directional chain elongation. A palladium-mediated, ligand-controlled desymmetrization provided the desired bis-oxanes in greater than 98% ee. Bis oxanes 1 and 4 represent potential synthetic intermediates for the C3-C17 subunits of phorboxazoles A and B. PMID- 14535743 TI - First examples of transition-metal free Sonogashira-type couplings. AB - [reaction: see text] We report here our observation that, using appropriate reaction conditions, the Sonogashira reaction can be performed without the need for transition-metal catalysts. Our approach involves the use of water as a solvent, poly(ethylene glycol) as a phase-transfer agent, and sodium hydroxide as a base. The methodology works, to differing extents, for aryl iodides and bromides. PMID- 14535745 TI - A convergent synthetic approach to the nucleoside-type liposidomycin antibiotics. AB - [reaction: see text] A synthetic approach toward the liposidomycins, a family of complex nucleoside-type antibiotics, is reported based on the synthesis of epoxy amides derived from the reaction of sulfur ylides with the uridyl aldehyde derivative 6. To this end, the epoxy-amide derivative of indoline 14 was stereoselectively prepared and, after treatment with DDQ, transformed into the corresponding N-indole epoxyamide 15. The indole 15 provides ready access to a variety of structures related to the diazepanone core present in the liposidomycins by reaction with a variety of amines. PMID- 14535744 TI - A novel three-component reaction catalyzed by dirhodium(II) acetate: decomposition of phenyldiazoacetate with arylamine and imine for highly diastereoselective synthesis of 1,2-diamines. AB - [reaction: see text] A practical highly diastereoselective synthesis of 1,2 diamines through carbon-carbon bond formation involving an ammonium ylide intermediate is reported for the first time. By treating methyl phenyldiazoacetate with arylamine and imine in the presence of dirhodium acetate, the erythro diastereomer of methyl 1,2-diaryl-1,2-diaminopropanoate is formed with stereochemical preferences greater than 10:1. PMID- 14535746 TI - First total synthesis of the 4a-methyltetrahydrofluorene diterpenoids (+/-) dichroanal B and (+/-)-dichroanone. AB - [reaction: see text] A simple synthesis of the 4a-methyltetrahydrofluorene diterpenoids (+/-)-dichroanal B and (+/-)-dichroanone has been achieved through a common hexahydrofluorenone intermediate obtained via Pd(0)-catalyzed reductive cyclization of a substituted 2-(2-bromobenzyl) methylene cyclohexane. PMID- 14535747 TI - Formal [3 + 3] cycloaddition approach to chromenes and chromanes. Concise total syntheses of (+/-)-rhododaurichromanic acids A and B and methyl (+/-) daurichromenic ester. AB - [reaction: see text] Total syntheses of (+/-)-rhododaurichromanic acids A and B and methyl (+/-)-daurichromenic ester are described here. Despite the complex appearances of these compounds, their syntheses are completed in six steps with a 15% overall yield as a mixture by featuring our formal oxa-[3 + 3] cycloaddition methodology. PMID- 14535748 TI - Absolute helical arrangement of sulfonamide in the crystal. AB - [reaction: see text] 1,2-Bis(N-benzenesulfonyl-N-methylamino)benzene (2), which has no fixed asymmetric element, was crystallized from ethyl acetate as chiral crystals belonging to space group P4(1)2(1)2 (No. 92) or P4(3)2(1)2 (No. 96). The array of molecules built by the CH-pi interaction along the c-axis forms an enantiomeric helical superstructure in each individual crystal. The absolute configurations of the chiral crystals of 2 were determined by X-ray crystal structure analysis using the Flack parameter method. The solid-state CD spectra of the chiral crystals in KBr were mirror images. The equilibrium between the two enantiomers in solution is fast during crystallization at ambient temperature, and the energy barrier (DeltaG()) is estimated to be 11.7 +/- 0.3 kcal/mol (233 K). PMID- 14535749 TI - Improved synthesis of meso-aryl-substituted [26]hexaphyrins. AB - [reaction: see text] Acid-catalyzed reactions of 5-(2',3',4',5',6' pentafluorophenyl)dipyrromethane with aryl aldehydes and of 5 (aryl)dipyrromethanes with 2,3,4,5,6-pentafluorobenzaldehyde allowed the preparation of a variety of meso-aryl [26]hexaphyrins. PMID- 14535750 TI - Autocatalytic asymmetric reduction of 2,6-diacetylpyridine. AB - [reaction: see text] We report here that the C(2)-symmetric diol 2,6-bis(1 hydroxyethyl)pyridine (2) can effect chiral-catalyzed reduction of 2,6 diacetylpyridine (1) and produce more of the diol (2) with the same configuration in an enantiomerically enriched form. The two carbonyl functionalities of (1) are reduced in 90% conversion to produce the enantio-enriched C(2)-symmetric diol (40% ee, 47% de) using zinc trifluoromethanesulfonate and a catalytic amount of the chiral C(2)-symmetric diol (2). PMID- 14535751 TI - Conformational and electronic engineering of twisted diphenylacetylenes. AB - [structure: see text] Three tethered diphenylacetylene derivatives were prepared by alkyne metathesis. In these cycles, the twist angle between the two benzene rings is variable and determined by the nature of the linker. The engineering of the twist angle leads to a change of the UV-vis spectra of the cycles. The larger the twist angle in the macrocycles, the more blue shifted their lambda(max) (UV vis), the lower their fluorescence quantum yield, and the lower field shifted their (13)C NMR signals of the alkyne carbons are. PMID- 14535752 TI - Toward transition metal-catalyzed carbonylation of methanol without HI as copromoter: catalytic exocyclic carbonylation of cycloimino esters. AB - [reaction: see text] Initial studies of a rare exocyclic C-O bond carbonylation are reported. Under the catalysis of Co(2)(CO)(8) in the absence of HI as the copromoter, cycloimino esters are carbonylated to afford N-acyllactams in high yields under relatively mild conditions (100-160 degrees C and 200-1000 psi). The reaction is interesting because it opens up the possibility of carbonylation of alcohols in the absence of HI. PMID- 14535753 TI - Cobalt-catalyzed three-component coupling reaction of alkyl halides, 1,3-dienes, and trimethylsilylmethylmagnesium chloride. AB - [reaction: see text] A combination of CoCl(2) and 1,6 bis(diphenylphosphino)hexane catalyzes a novel three-component coupling reaction of alkyl bromides, 1,3-dienes, and silylmethylmagnesium chloride, yielding homoallylsilanes in good to excellent yields. The reaction involves a radical species from alkyl halides. PMID- 14535754 TI - Cobalt-catalyzed carbocyclization of o-iodobenzaldehydes and o-iodophenylketones with alkynes. AB - [reaction: see text] Treatment of various o-iodobenzaldehydes and o-iodophenyl ketones with alkynes in the presence of Co(dppe)I(2) and Zn powder in acetonitrile at 80 degrees C afforded the corresponding indenols in moderate to excellent yields with exceedingly high regioselectivity. For most unsymmetrical alkynes tested, the carbocyclization gave a single regioisomer. PMID- 14535755 TI - Water exclusion reaction in aqueous media: nitrone formation and cycloaddition in a single pot. AB - [reaction: see text] The formation of nitrone (a water exclusion reaction) in aqueous media using surfactant and subsequent cycloaddition in the same pot, a new example of green chemistry, is reported. The control of regioselectivity favors the formation of trans-5-substituted isoxazolidine. This work not only may lead to an environmentally benign system but also will provide a new aspect of reactions in water. PMID- 14535756 TI - Stereoselective synthesis of beta-(chloro)vinylsilanes using a regio- and (E) stereoselective bis-stannylation of unsymmetrically substituted butadiynes: application to the synthesis of a masked triyne. AB - [reaction: see text] A highly regio- and stereoselective bis-stannylation of unsymmetrically substituted butadiyne 3 provides bis-stannane 4. Selective lithiation of the internal tin residue effects a 1,4-retro-Brook rearrangement to afford vinylsilane 5. This was elaborated into the novel diethynylethene 1, which also functions as a masked triyne. PMID- 14535757 TI - Rapid and efficient synthesis of 1H-indol-2-yl-1H-quinolin-2-ones. AB - [reaction: see text] A concise and efficient synthesis of the novel indol-2-yl-1H quinolin-2-one ring system found in the potent and selective KDR kinase inhibitors 1-3 is presented. PMID- 14535758 TI - Useful enantioselective bicyclization reactions using an N-protonated chiral oxazaborolidine as catalyst. AB - [reaction: see text] Nine examples are reported of enantioselective [4 + 2] cycloaddition reactions of achiral, acyclic substrates to form chiral bicyclo[4.3.0]nonane or bicyclo[4.4.0]decane derivatives. PMID- 14535759 TI - Versatile route to centro-substituted triquinacene derivatives: synthesis of 10 phenyltriquinacene. AB - [reaction: see text] A versatile route to prepare centro-substituted triquinacene derivatives (1, R = various substituents), as exemplified by the preparation of 10-phenyltriquinacene (1, R = Ph), is reported. The quaternary, centro substituent (C-10) was installed by a trimethylsilyl chloride-promoted conjugate addition reaction of an organocuprate, derived from phenylmagnesium bromide, and the protected bicyclic enone (11). The resultant trimethylsilyl enol ether was then converted regioselectively to the C-2-allylated conjugate addition products (13, R = Ph). The allyl moiety, following oxidative cleavage of the carbon-carbon double bond, was used to elaborate the tricyclic ring system by an intramolecular aldol/acetal deprotection reaction. The product of this reaction was then converted to the target compound using a standard series of functional group transformation reactions. PMID- 14535760 TI - Asymmetric sulfur ylide mediated aziridination: application in the synthesis of the side chain of taxol. AB - [reaction: see text] Sulfur ylide methodology has been used to construct the Taxol side chain with a high degree of enantioselectivity via a trans-aziridine followed by stereospecific rearrangement of the trans-benzoylaziridine into a trans-oxazoline. PMID- 14535761 TI - Sieboldine A, a novel tetracyclic alkaloid from Lycopodium sieboldii, inhibiting acetylcholinesterase. AB - [structure: see text] A novel Lycopodium alkaloid with an unprecedented fused tetracyclic ring system consisting of an aza-cyclononane ring having a N-hydroxy group, a cyclohexanone, a cyclopentanone, and a tetrahydrofuran ring, sieboldine A (1), was isolated from the club moss Lycopodium sieboldii. The structure and relative stereochemistry were elucidated on the basis of 2D NMR data and X-ray analysis. Sieboldine A (1) exhibited a potent inhibitory activity against acetylcholinesterase and modest cytotoxicity. PMID- 14535762 TI - Enantioselective [4 + 2]-annulation of chiral crotylsilanes: application to the synthesis of a C1-C22 fragment of leucascandrolide A. AB - [reaction: see text] The asymmetric synthesis of a C1-C22 fragment (2) of leucascandrolide A is described. Synthetic highlights include the construction of the C9-C22 pyran fragment using a formal [4 + 2]-annulation of a chiral organosilane. A diastereoselctive Mukaiyama aldol was used to introduce the C9 stereocenter and complete the assembly of the macrocycle's carbon skeleton. PMID- 14535763 TI - Total synthesis of 3',5'-C-branched nucleosides. AB - [reaction: see text] A novel total synthesis of 3',5'-C-branched uridine azido acid has been accomplished using stereoselective aldehyde alkynylation, Ireland Claisen rearrangement, and iodolactonization as the key reactions. Compared to traditional routes that start from carbohydrates, the present methodology is more efficient, flexible for future optimization, and provides access to both enantiomers of the products. Because the key chemistry does not involve the 3'- and 5'-C substituents, our route is a general approach to 3',5'-C alkyl nucleoside analogues. PMID- 14535764 TI - Molecular recognition of anions through hydrogen bonding stabilization of anion ionophore adducts: a novel trifluoroacetophenone-based binding motif. AB - [reaction: see text] A novel trifluoroacetophenone-based binding motif has been developed that recognizes anions such as carboxylates through reversible formation of anion-ionophore adducts that are stabilized by intramolecular H bonding. The intramolecular H-bonding resulted in more than 10-fold enhancement in the binding affinity and an enthalpy gain (DeltaH degrees ) of 3.0 kcal/mol for the binding of an acetate ion when compared to the case without the intramolecular H-bonding. PMID- 14535765 TI - Enantiospecific formal total syntheses of (-)-salicylihalamides A and B from D glucose and L-rhamnose. AB - [reaction: see text] Two formal chiral pool syntheses of the (-) salicylihalamides A and B were achieved from commercially available 1,2,5,6 diacetone-d-glucose and l-rhamnose. PMID- 14535766 TI - Copper-mediated N-alkynylation of carbamates, ureas, and sulfonamides. A general method for the synthesis of ynamides. AB - [reaction: see text] A general amination strategy for the N-alkynylation of carbamates, sulfonamides, and chiral oxazolidinones and imidazolidinones is described. A variety of substituted ynamides are available by deprotonation of amides with KHMDS followed by reaction with CuI and an alkynyl bromide. PMID- 14535768 TI - Aggregation and reactivity of the dilithium and dicesium enediolates of 1 naphthylacetic acid. AB - UV-vis spectra of the dilithium, 1-Li, and dicesium, 1-Cs, enediolates of alpha naphthylacetic acid show no systematic change with concentration in dilute THF solution, but addition of small amounts of HMPA causes a bathochromic shift in the spectrum of 1-Li. These results indicate that these salts are aggregated and that HMPA breaks up the aggregates of 1-Li. The quantitative effect of small increments of HMPA indicates that 1-Li is a dimer. Alkylation reactions of 1-Cs show half-order kinetics in enediolate indicating that this salt is also dimeric but that the small amount of monomer in equilibrium is the actual reactant. Alkylation of 1-Li, however, is much slower and shows first-order kinetics interpreted as a direct reaction of the dimer; the amount of monomer in this case is too small to compete. A solution of 1-Li in THF containing 10% HMPA is much more reactive in alkylation than 1-Li alone and the first-order dependence in 1 Li is now interpreted as reaction of the monomer. Compound 1-Li is found to form a mixed aggregate with LDA, a finding that has possible synthetic significance since enediolates used in syntheses are frequently prepared using LDA. Structures of these compounds are suggested based on model ab initio computations. PMID- 14535769 TI - Spiro-fused (C2)-azirino-(C4)-pyrazolones, a new heterocyclic system. Synthesis, spectroscopic studies and X-ray structure analysis. AB - Reaction of 1-substituted 4-acyl-5-hydroxy-3-methyl-1H-pyrazoles (2) with hydroxylamine gives the corresponding "oximes" 3, which are mainly present as (Z) 2,4-dihydro-4-[(hydroxyamino)methylene]-3H-pyrazol-3-ones. Treatment of compounds 3 with trichloroacetyl isocyanate/potassium carbonate in anhydrous diethyl ether affords 7-methyl-1,5,6-triazaspiro[2.4]hepta-1,6-dien-4-ones (4). The structure of compounds 4 was elucidated by means of single-crystal X-ray analysis (4f, 4h) and confirmed by NMR spectroscopic investigations ((1)H, (13)C). PMID- 14535770 TI - Photolysis of alpha-azidoacetophenones: direct detection of triplet alkyl nitrenes in solution. AB - We report the first detection of triplet alkyl nitrenes in fluid solution by laser flash photolysis of alpha-azido acetophenone derivatives, 1. Alphazides 1 contain an intramolecular triplet sensitizer, which ensures formation of the triplet alkyl nitrene by bypassing the singlet nitrene intermediate. At room temperature, azides 1 cleave to form benzoyl and methyl azide radicals in competition with triplet energy transfer to form triplet alkyl nitrene. The major photoproduct 3 arises from interception of the triplet alkyl nitrene with benzoyl radicals. The triplet alkyl nitrene intermediates are also trapped with molecular oxygen to yield the corresponding 2-nitrophenylethanone. Laser flash photolysis of 1 reveals that the triplet alkyl nitrenes have absorption around 300 nm. The triplet alkyl nitrenes were further characterized by obtaining their UV and IR spectra in argon matrices. (13)C and (15)N isotope labeling studies allowed us to characterize the C-N stretch of the nitrene intermediate at 1201 cm(-)(1). PMID- 14535771 TI - Two novel alkaloids with a unique fused hexacyclic skeleton from Daphniphyllum subverticillatum. AB - Two novel major alkaloids, deoxycalyciphylline B (1) and deoxyisocalyciphylline B (2) with a unique fused hexacyclic skeleton, together with a quite recently reported alkaloid calyciphylline B (3), were isolated from the stem of Daphniphyllum subverticillatum. Their structures were established by spectral methods and chemical evidence, especially 2D NMR techniques. The structure of 1 was further confirmed by a single-crystal X-ray diffraction determination. PMID- 14535772 TI - Regulated-stereoselective construction of thirteen stereogenic centers necessary for the frame of (+)-discodermolide, based on iterative Lewis acid-promoted aldol reactions. AB - The segments C(1)-C(13) and C(15)-C(21) containing the 13 stereogenic centers required for the frame of (+)-discodermolide were synthesized in good to excellent enantio- and diastereoselectivities from a common racemic aldehyde, derived from 2-methyl-1,3-propanediol. The enantioselective aldol reactions of the racemic aldehyde with a silylketene acetal, derived from ethyl 2 bromopropionate, in the presence of chiral oxazaborolidinones, prepared in situ with N-p-toluenesulfonyl-(R)- and -(S)-valine and BH(3).THF, proceeded under kinetic control to give the stereotriads with a high degree of enantioselectivity. Enantioselective (chiral borane) and diastereoselective (BF(3).OEt(2) and TiCl(4)) aldol reactions with the silylketene acetal, coupled with diastereoselective radical debrominations (Bu(3)SnH, Et(3)B, with or without MgBr(2)), were used iteratively. This aldol reaction strategy for the construction of the polypropionate frame dramatically shortened the steps needed for the construction of the final segments. PMID- 14535773 TI - Highly efficient Peterson olefination leading to unsaturated selenoamides and their characterization. AB - The Peterson olefination of aromatic aldehydes with an alpha-silyl selenoacetamide proceeded smoothly with high stereoselectivity to give E alpha,beta-unsaturated selenoamides in good to high yields. The reaction with aldehydes bearing alkenyl and dienyl groups gave the corresponding selenoamides bearing dienyl and trienyl groups, but the stability of the products depended on the substituents on the aromatic ring. X-ray molecular analysis disclosed that the alpha,beta,gamma,delta-unsaturated selenoamides had a nearly planar structure. In the (77)Se NMR spectra, signals were observed in the region greater than 130 ppm depending on the substituents on the aromatic ring, whereas the (1)J coupling constant between the carbon atom and the selenium atom was almost independent of the substituents. A linear relationship was observed between the chemical shifts in the (77)Se NMR spectra and Hammet sigma parameters, and this correlation was retained even when one or two alkenyl groups were introduced to alpha,beta-unsaturated selenoamides, although it became less sensitive. PMID- 14535775 TI - Complete and remarkable reversal of chemoselectivity in [4 + 2] cycloadditions involving electron-poor indoles as dienophiles. Diels-Alder versus hetero-Diels Alder processes. AB - The reaction between indole-3-carboxaldehyde 1a or indole-3-glyoxalate 1b and 2,3 dimethylbutadiene under thermal activation leads exclusively to the Diels-Alder cycloadducts resulting from the participation of the indole 2,3-carbon-carbon double bond. The concomitant use of zinc chloride and high pressure (16 kbar) induces the primary cycloadduct to react further, and biscycloadducts 11 and 12 are now isolated in high yields, the result of two consecutive [4 + 2] processes on, first, the indole 2,3 C=C bond and, second, the 3-carbonyl unit. The possibility of using two different dienes in a tandem, sequential process is demonstrated by the preparation of tetracycle 13. Interactions between the carbonyl dienophile and Danishefsky diene yield exclusively yet another type of product, namely the gamma-dihydropyranones arising from the sequential [4 + 2] heterocycloaddition, hydrolysis of the silyl enol ether, and loss of methanol. Isolation of the Mukaiyama-type adduct 16 indicates that a stepwise mechanism may be involved, at least under zinc chloride catalysis. N,N-Disubstituted indole-3 glyoxamides undergo the expected, usual Diels-Alder process, with the 2,3 C=C bond acting as dienophile, and cycloadducts of the type 3 are obtained in high yields, regardless of the mode of activation. Remarkably, however, N monosubstituted indole-3-glyoxamides react almost exclusively as heterodienophiles, the 3-carbonyl unit being now the preferred site of reactivity, and gamma-dihydropyranones of the type 6 are isolated in yields ranging from 72 to 92%. Conformational analysis of the Diels-Alder adducts based on both (1)H NMR spectrometry and X-ray diffraction data indicates that the newly created cyclohexene and cyclohexanone rings adopt a pseudoboat conformation. PMID- 14535774 TI - Total synthesis of mappicine ketone (nothapodytine B) by means of sulfur-directed 5-exo-selective aryl radical cyclization onto enamides. AB - Enamides 5, on treatment with Bu(3)SnH-AIBN, underwent aryl radical cyclization in a 5-exo manner to give 1-[bis(phenylthio)methyl]dihydroisoindoles 6, which were partially desulfurized with Bu(3)SnH-AIBN to give 1-mono(phenylthio)methyl congeners 7. Formation of 6 from 5 may be explained by the presence of two phenylthio groups at the terminus of the N-vinylic bond of 5, since enamide 8a having no phenylthio group underwent aryl radical cyclization in a 6-endo manner. Compound 7d (R = CF(3)) was transformed into sulfoxide 16, which was treated with (CF(3)CO)(2)O and then with 10% NaOH to give a model compound 20 of mappicine ketone (MPK) (1) through aldol condensation of aldehyde 18. An attempt to synthesize MPK using this method with sulfoxide 28 prepared from 25, however, was unsuccessful, and, instead, photochemical cyclization of enamide 38 prepared from 25 furnished MPK. PMID- 14535776 TI - P(i-BuNCH2CH2)3N: an efficient ligand for the direct alpha-arylation of nitriles with aryl bromides. AB - A new catalyst system for the synthesis of alpha-aryl-substituted nitriles is reported. The bicyclic triaminophosphine P(i-BuNCH(2)CH(2))(3)N (1b) serves as an efficient and versatile ligand for the palladium-catalyzed direct alpha-arylation of nitriles with aryl bromides. Using ligand 1b, ethyl cyanoacetate and primary as well as secondary nitriles are efficiently coupled with a wide variety of aryl bromides possessing electron-rich, electron-poor, electron-neutral, and sterically hindered groups. PMID- 14535777 TI - Synthesis of aryl- and heteroaryl[a]pyrrolo[3,4-c]carbazoles. AB - Synthesis of aryl- and hetero[a]pyrrolo[3,4-c]carbazoles by photochemical oxidation and Heck cyclization are described. Photochemical oxidation of 2 naphthyl indolyl maleimide affords two different carbazole regioisomers, depending on the reaction conditions. The regiochemistry of the cyclization can be controlled using the Heck reaction. PMID- 14535778 TI - Tandem double intramolecular [4+2]/[3+2] cycloadditions of nitroalkenes. The fused/bridged mode. AB - A new class of tandem [4+2]/[3+2] cycloadditions of nitroalkenes is described in which both pericyclic processes are intramolecular. Two subclasses of intra [4+2]/intra [3+2] cycloadditions have been explored in which the dipolarophile is tethered at either C(5) or C(6) of the nitronate. For both families of precursors, the cycloadditions occur in good yield and are found to be highly regio- and stereoselective. This method converts linear polyenes to functionalized polycyclic systems bearing up to six stereogenic centers. PMID- 14535779 TI - Spin exchange interaction through phenylene-ethynylene bridge in diradicals based on iminonitroxide and nitronylnitroxide radical derivatives. 1. Experimental investigation of the through-bond spin exchange coupling. AB - A series of bis-iminonitroxide diradical derivatives of different lengths and geometry have been prepared that incorporate a conjugated phenylene-ethynylene bridge as a rigid spacer. This paper describes the synthesis of these new components and their main characterizations. An unexpected singlet ground state and substituent effects on the singlet-triplet gap have been found for substituted "m-phenylene"-based diradicals. The effects of the pi-conjugation on the intramolecular through-bond spin coupling have been investigated by changing the length of the spacer within linear derivatives. The EPR studies demonstrate the intramolecular magnetic coupling between the radical spins within all compounds. This result is very attractive and unusual, given the large distance between the radicals from 15 A in the dimer to 36 A in the pentamer. PMID- 14535780 TI - Asymmetric synthesis of fused bicyclic alpha-amino acids having a hexahydro cyclopenta[c]pyridine skeleton via intramolecular Pauson-Khand reaction of 1 sulfonimidoyl-substituted 5-azaoct-1-en-7-ynes. AB - An asymmetric synthesis of fused bicyclic amino acids having a hexahydro cyclopenta[c]pyridine skeleton and carrying besides an enone structural element a substituent at the beta-position is described. The key steps of the synthesis are a highly selective allylation of N-tert-butylsulfonyl imino ester with bis(allylsulfoximine)titanium complexes and a highly diastereoselective Pauson Khand cycloaddition of sulfonimidoyl-substituted gamma,delta-unsaturated alpha amino acid esters carrying a substituent at the beta-position and a propargyl group at the N-atom. The cyclization is accompanied by a reductive cleavage of the sulfoximine group of the primary cyclization product. Surprisingly, the removal of the sulfoximine group proceeds with inversion of the configuration at the S-atom and gives N-methyl-phenylsulfinamide with >/=98% ee. Deprotection of the bicyclic N-tert-butylsulfonyl-protected amino acid ester was accomplished through treatment with CF(3)SO(3)H under anhydrous conditions. The enantio- and diastereomerically pure sulfoximine-substituted gamma,delta-unsaturated alpha amino acid esters used as starting material were obtained through a highly regio- and diastereoselective allylation of N-tert-butylsulfonyl imino ester with acyclic bis(allylsulfoximine)titanium complexes, described previously. PMID- 14535781 TI - Synthesis of both possible isomers of the northwest quadrant of altromycin B. AB - The synthesis of the northwest quadrant of Altromycin B is described. The preparation of the two epimers at the quaternary carbon of the 6-deoxy-C-altrose moiety in the northwest quadrant is accomplished starting from d-glucose. A key step of our synthetic sequence is the formation of the C-glycoside linkage via the Ramberg-Backlund reaction. Two different routes are explored, which differ mainly on the timing of the conversion of glucose to altrose, either before or after the preparation of the C-glycoside. The conformation behavior of variously substituted C-altropyranoside rings is also discussed. PMID- 14535782 TI - Asymmetric synthesis of the quinolizidine alkaloid (-)-epimyrtine with intramolecular Mannich cyclization and N-sulfinyl delta-amino beta-ketoesters. AB - A concise, six-step, enantioselective synthesis of (-)-epimyrtine employing the N sulfinyl delta-amino beta-ketoester chiral building block is described. PMID- 14535783 TI - Rearrangements of bicyclic nitrones to lactams: comparison of photochemical and modified Barton conditions. AB - The rearrangement of nitrones to lactams can be carried out by photochemical activation or by treatment with Tf(2)O followed by KOH-promoted rearrangement (a modification of conditions originally introduced by Barton). Substrates in which the nitrone is part of a fused bicyclic ring system have traditionally proven problematic for this kind of reaction. In this study, a series of mono-, bi-, and tricyclic ring-fused nitrones were prepared to investigate the dependence of products on nitrone ring size and tether length. Results indicated that photochemical rearrangement of nitrones in benzene afforded reasonably good yields (30-68%) of lactams, while the two-step nonphotochemical process provided slightly better average yields (30-95%) of the same targets. PMID- 14535784 TI - Novel synthesis of carbapenam by intramolecular attack of lactam nitrogen toward eta1-allenyl and eta3-propargylpalladium complex. AB - When a THF solution of beta-lactam having propargyl phosphate was warmed in the presence of 5 mol % of Pd(2)(dba)(3) x CHCl(3), 20 mol % of a bidentate ligand, and sodium acetate (1.5 equiv) at 40 degrees C for 22 h, carbapenam was produced in high yield. In this reaction, the lactam nitrogen attacked the central carbon of a eta(3)-propargylpalladium complex, which was formed from propargyl phosphate and Pd(0). PMID- 14535786 TI - Lasonolide A: structural revision and total synthesis. AB - The proposed structure of lasonolide A was synthesized employing radical cyclization reactions of beta-alkoxyacrylates for preparation of the tetrahydropyranyl units A and B, but the spectroscopic data did not match those of the natural product. Both enantiomers of a revised structure featuring 17E,25Z double bonds were synthesized, and the (-)-isomer was found to be the biologically active enantiomer. PMID- 14535785 TI - Synthesis of a tritium-labeled, fipronil-based, highly potent, photoaffinity probe for the GABA receptor. AB - 3-[4-[1-(2,6-dichloro-4-trifluoromethylphenyl)pyrazolo]]-3 (trifluoromethyl)diazirine is a fipronil-based (i.e. fiprole), high-affinity probe for the GABA receptor. For synthesis of the tritium-labeled version of this trifluoromethyldiazirinylfiprole ([(3)H]TDF) the required intermediate, 3-[4-[1 (2,6-dichloro-3-iodo-4-trifluoromethylphenyl)-5-iodopyrazolo]]-3 (trifluoromethyl)diazirine, was prepared in 10 steps from pyrazole and 3,5 dichloro-4-fluorobenzotrifluoride. One of the key transformations was lithiation and subsequent iodination of the 4-(2,2,2-trifluoro-1-hydroxyethyl)pyrazole intermediate. The last step involved reduction of the diiodofiprole with tritium, Pd/C, and triethylamine in ethyl acetate and afforded [(3)H]TDF with a specific activity of 15 Ci/mmol and 99% radiopurity. PMID- 14535787 TI - Asymmetric synthesis of cyclic hydroxy ketones derived from enol ethers via sharpless asymmetric dihydroxylation. A study in the correlation of the enol ether chain length and enantioselectivity. AB - The Sharpless asymmetric dihydroxylation reaction of enol ethers derived from their corresponding cyclic ketones, gave alpha-hydroxyketones with high enantioselectivity. The enantiomeric excess was found to be proportional to the length of the unbranched enol ether chain with a maximum ee for the pentyl enol ether. An efficient synthesis of alpha-hydroxy chromanone in >90% ee was demonstrated using this method. PMID- 14535788 TI - General method for the palladium-catalyzed allylation of aliphatic alcohols. AB - A palladium catalysis-mediated approach to coupling aliphatic alcohols with allyl carbonates has been developed. The method allows for the allylation of primary, secondary, and tertiary alcohols efficiently under mild conditions. Limitations were explored as well as the asymmetric application of the chemistry. Regiochemical and olefin geometry was controlled in the coupling of unsymmetrical allylating agents. Transient allyl carbonates were observed in the coupling, which comprised the trans-carboxylation of the allyl-carbonate with the requisite alcohol. PMID- 14535789 TI - Epimerization of 2'-carbonylalkyl-C-glycosides via enolation, beta-elimination and intramolecular cycloaddition. AB - Treatment of 2'-carbonyl-alpha-C-glycopyranosides of gluco, galacto, manno, 2 deoxy, and 2-azido sugars with 4% NaOMe resulted in anomeric epimerization to give their respective beta-anomers in good to excellent yields. The epimerization of the 2'-aldehyde of alpha-C-galactopyranoside (10) in deuterium methanol, which afforded the beta-anomer with exclusive deuterium replacements at the 1' position, excluded the possibility of the exo-glycal as being involved as an intermediate. When 2'-aldehyde (36) and 2'-ketone (41) of 2,3-di-O-benzyl alpha/beta-l-C-arabinofuranoside were used as substrates we were able to obtain the respective equatorial alpha-C-arabinopyranosides (37 and 42). These observations confirmed that the epimerization involves an acyclic alpha,beta unsaturated aldehyde or ketone, which is formed by the enolation of 2'-carbonyl alpha-C-glycoside with subsequent beta-elimination. Thereafter an intramolecular hetero-Michael cycloaddition occurs, leading to the formation of thermodynamically controlled stable products, which were exclusively the equatorial C-glycopyranosides, except in the case of 2'-carbonyl-C-furanosides, where a mixture of two anomers was obtained. PMID- 14535790 TI - Application of silicon-based cross-coupling technology to triflates. AB - Aryl silatranes undergo fluoride-induced cross-coupling with aryl triflates to provide unsymmetrical biaryl derivatives in good to excellent yields. Silatranes also couple with aryl iodides and bromides, although the yields of adduct are lower than with the corresponding siloxane derivates. Aryl siloxanes (which had previously failed to couple with triflates) can be employed for triflate couplings using the Denmark modification, although the yields are lower than the corresponding silatrane reactions. PMID- 14535791 TI - Synthesis, electrochemistry, and gas-phase photoelectron spectroscopic and theoretical studies of 3,6-bis(perfluoroalkyl)-1,2-dithiins. AB - 3,6-bis(trifluoromethyl)- and 3,6-bis(pentafluoroethyl)-1,2-dithiin (1a,b), the first known perfluoroalkyl-substituted 1,2-dithiins, were synthesized from (Z,Z) 1,4-bis(tert-butylthio)-1,3-butadiene (2) to evaluate the effects of electron withdrawing groups on the ionization and oxidation potentials of 1,2-dithiins. Analysis of the photoelectron spectra of 1a and 1b provided a basis for assigning orbital compositions. Ab initio calculations on these compounds showed that they adopt a twist geometry as does 1,2-dithiin (1c) itself. Cyclic voltammetric studies on 1a and 1b revealed a reversible oxidation followed by an irreversible oxidation at much more positive potentials than for 1,2-dithiin and 3,6-dimethyl 1,2-dithiin (1d). The oxidation potentials determined electrochemically do not correlate with the ionization potentials determined by photoelectron spectroscopy. This result supports the previously advanced hypothesis that there is a geometry change on electrochemical oxidation leading to a planar radical cation. PMID- 14535792 TI - An efficient route for the synthesis of glycosyl phosphinic acids. AB - An efficient method for the synthesis of glycosyl phosphinic acids (21) from the corresponding C-phosphonates is described. The route developed involves three steps: reduction of the glycosyl C-phosphonate to a primary phosphine, reaction of this product with an alkylating agent to afford a secondary phosphine, and finally oxidation to the phosphinic acid. Deprotection provides compounds suitable for testing as glycosyl phosphate analogues. Although the focus of this report is the synthesis of analogues of arabinofuranosyl-containing phosphate esters, the method should be readily applicable to other systems, carbohydrate or otherwise. PMID- 14535793 TI - Synthesis of nanostructures based on 1,4- and 1,3,5-ethynylphenyl subunits with pi-extended conjugation. Carbon dendron units. AB - Nanometer-sized conjugated 1,4- and 1,3,5-ethynylphenyl oligomers were synthesized starting from 3,5-di(trimethylsilylethynyl)phenylacetylene and p-[3,5 di(trimethylsilylethynyl)-1-ethynylphenyl]phenyl acetylene by cross-coupling reaction with a convenient haloaryl derivative, catalyzed by palladium(II)/copper(I), in excellent yield. The terminal acetylenes were efficiently prepared by a specific protection-deprotection methodology. All ethynylphenyl homologues obtained show fluorescence emission, with the bathochromic shift of approximately 20 nm by each ethynylphenyl unit increasing the conjugate chain. Parallel conjugated ethynylphenyl chains were prepared through the insertion of a 1,5-naphthalene subunit, and the compounds exhibit fluorescence radiation emission. PMID- 14535794 TI - A formal total synthesis of the salicylihalamides. AB - The synthesis of the macrolactone 23 is described. The synthesis features a diastereoselective hydroboration of the chiral alkene 17 followed by a Suzuki cross-coupling reaction with the benzoate 5. The resulting seco acid 21 was converted to the macrolactone 23 by a Mitsunobu lactonization using immobilized triphenylphosphine. The stereogenic centers in the alkene 17 were established by a Noyori reduction of the beta-keto ester 8 and an Evans aldol reaction. The synthesis illustrates the conversion of a syn aldol product to the corresponding anti product by inversion of the methyl-bearing center. PMID- 14535795 TI - Using a lipase as a high-throughput screening method for measuring the enantiomeric excess of allylic acetates. AB - This report describes a high-throughput method for measuring the enantiomeric excess of allylic acetates. Such methods are useful tools for screening libraries of potential catalysts for enantioselective reactions. This technique, which is called EMDee for an enzymatic method for determining enantiomeric excess, uses the lipase from Pseudomonas cepacia to hydrolyze the (R) enantiomer of an allylic acetate, while the (S) enantiomer does not react. The rate of the reaction is monitored by measuring the acetic acid that is produced during the hydrolysis reaction with a pH indicator. Using the Michaelis-Menten equation, the rate of the reaction can be correlated with the concentration of the (R) enantiomer. This method can process 88 samples in less that 30 min. PMID- 14535796 TI - Influence of the 4,6-O-benzylidene, 4,6-O-phenylboronate, and 4,6-O polystyrylboronate protecting groups on the stereochemical outcome of thioglycoside-based glycosylations mediated by 1-benzenesulfinyl piperidine/triflic anhydride and N-iodosuccinimide/trimethylsilyl triflate. AB - The effect of 4,6-O-benzylidene acetals, 4,6-O-phenylboronate esters, and 4,6-O polystyrylboronate esters on the stereoselectivity of couplings to galacto-, gluco-, and mannopyranosyl thioglycosides, otherwise protected with benzyl ethers, has been investigated by the benzenesulfinyl piperidine/trifluoromethanesulfonic anhydride (BSP), diphenyl sulfoxide/trifluoromethanesulfonic anhydride (Ph(2)SO), and N iodosuccinimide/trimethylsilyl trifluoromethanesulfonate (NIS/TMSOTf) methods. The BSP and Ph(2)SO methods give comparable results in all three systems whereas the NIS method affords significantly different stereoselectivities in both the gluco and manno, but not the galacto series. The benzylidene acetal and boronate esters influence the stereochemistry in a similar manner in the beta-selective manno series and the alpha-selective galacto series but show significant differences with the glucose donors. The differences between the glucose, galactose, and mannose series reflect the established differences in reactivity and, especially for mannose, those in the anomeric effect and are best interpreted in terms of changes in the relative energetics between the alpha- and beta-covalent triflate intermediates and the various contact ion pairs with which they are necessarily in equilibrium. PMID- 14535797 TI - Binding of tetramethylammonium to polyether side-chained aromatic hosts. Evaluation of the binding contribution from ether oxygen donors. AB - A set of macrocyclic and open-chain aromatic ligands endowed with polyether side chains has been prepared to assess the contribution of ether oxygen donors to the binding of tetramethylammonium (TMA), a cation believed incapable of interacting with oxygen donors. The open-chain hosts consisted of an aromatic binding site and side chains possessing a variable number of ether oxygen donors; the macrocyclic ligands were based on the structure of a previously investigated host, the dimeric cyclophane 1,4-xylylene-1,4-phenylene diacetate (DXPDA), implemented with polyether-type side chains in the backbone. Association to tetramethylammonium picrate (TMAP) was measured in CDCl(3) at T = 296 K by (1)H NMR titrations. Results confirm that the main contribution to the binding of TMA comes from the cation-pi interaction established with the aromatic binding sites, but they unequivocally show that polyether chains participate with cooperative contributions, although of markedly smaller entity. Water is also bound, but the two guests interact with aromatic rings and oxygen donors in an essentially noncompetitive way. An improved procedure for the preparation of cyclophanic tetraester derivatives has been developed that conveniently recycles the oligomeric ester byproducts formed in the one-pot cyclization reaction. An alternative entry to benzylic diketones has also been provided that makes use of a low-order cyanocuprate reagent to prepare in fair yields a class of compounds otherwise uneasily accessible. PMID- 14535798 TI - Kinetic investigation of the reactions of S-4-nitrophenyl 4-substituted thiobenzoates with secondary alicyclic amines in aqueous ethanol. AB - The reactions of S-4-nitrophenyl 4-X-substituted thiobenzoates (X = H, Cl, and NO(2): 1, 2, and 3, respectively) with a series of secondary alicyclic amines (SAA) were subjected to a kinetic investigation in 44 wt % ethanol-water, at 25.0 degrees C and an ionic strength of 0.2 M (KCl). The reactions were followed spectrophotometrically by monitoring the release of 4-nitrobenzenethiolate anion at 420-425 nm. Under excess amine, pseudo-first-order rate constants (k(obsd)) are obtained for all reactions. The plots of k(obsd) vs [SAA] at constant pH are linear with the slope (k(N)) independent of pH. The statistically corrected Bronsted-type plots (log k(N)/q vs pK(a) + log p/q) for the reactions of 1 and 2 are nonlinear with slopes at high pK(a), beta(1) = 0.27 and 0.10, respectively, and slopes at low pK(a), beta(2) = 0.86 and 0.84, respectively. The Bronsted curvature is centered at pK(a) (pK(a)(0)) 10.0 and 10.4, respectively. The reactions of SAA with 3 exhibit a linear Bronsted-type plot of slope 0.81. These results are consistent with a stepwise mechanism, through a zwitterionic tetrahedral intermediate (T(+/-)). For the reactions of 1 and 2, there is a change in rate-determining step with amine basicity, from T(+/-) breakdown to products at low pK(a), to T(+/-) formation at high pK(a). For the reactions of 3, breakdown to products of T(+/-) is rate limiting for all the SAA series (pK(a)(0) > 11). The increasing pK(a)(0) value as the substituent in the acyl group becomes more electron withdrawing is attributed to an increasing nucleofugality of SAA from T(+/-). The greater pK(a)(0) value for the reactions of SAA with 1, relative to that found in the pyridinolysis of 2,4-dinitrophenyl benzoate (pK(a)(0) = 9.5), is explained by the greater nucleofugality from T(+/-) of the former amines, compared to isobasic pyridines, and the greater leaving ability from T(+/ ) of 2,4-dinitrophenoxide relative to 4-nitrobenzenethiolate. PMID- 14535799 TI - Total synthesis of (+)-geldanamycin and (-)-o-quinogeldanamycin: asymmetric glycolate aldol reactions and biological evaluation. AB - The total synthesis of (+)-geldanamycin (GA), following a linear route, has been completed using a demethylative quinone-forming reaction as the last step. Key steps include the use of two new asymmetric boron glycolate aldol reactions. To set the anti-C11,12 hydroxymethoxy functionality, (S,S)-5,6-bis-4 methoxyphenyldioxanone 8 was used. Methylglycolate derived from norephedrine 5 set the C6,7 methoxyurethane stereochemistry. The quinone formation step using nitric acid gave the non-natural o-quino-GA product 55 10:1 over geldanamycin. Other known oxidants gave an unusual azaquinone product 49. o-Quino-GA 55 binds Hsp90 with good affinity but is less cytotoxic compared to GA. PMID- 14535800 TI - A theoretical study of the mechanism of selective fluorination of saturated hydrocarbons by molecular fluorine. Participation of CHCl3 solvent molecules in the ionic process. AB - The fluorination reaction of methane and isobutane by molecular F(2) with and without CHCl(3) solvent was studied with the ab initio and ONIOM theoretical calculations. The electrophilic pathway for the RH + F(2) reaction in general is a two-step process, consisting of hydride abstraction leading to an intermediate of the type R(+)(delta)...HF...F(-)(delta), followed by complicated rearrangement to give the electrophilic substitution product, RF + HF. In the case of methane, the overall barrier for this reaction is too high for the reaction to take place under mild conditions even in the presence of CHCl(3) solvent molecules. In the case of isobutane without CHCl(3), the electrophilic pathway has a high rate determining barrier of 25 kcal/mol, and is not likely to take place; the radical process forming t-Bu* + HF + F* may be preferred. However, the t Bu(+)(delta)...HF...F(-)(delta) intermediate and, in particular, the transition state TS2 for rearrangement of the intermediate are highly ionic, and are stabilized dramatically when a few CHCl(3) solvent molecules form a solvation cage. The electrophilic reaction for isobutane + F(2) has a low overall barrier when at least three CHCl(3) molecules are present and can take place under mild conditions with full retention of configuration. PMID- 14535801 TI - Diastereoselective and enantioselective cyclopropanation of alkenes catalyzed by cobalt porphyrins. AB - Cobalt(II) porphyrin complexes were shown to be general and efficient catalysts for selective cyclopropanation of alkenes with ethyl diazoacetate (EDA). The catalytic system can operate with alkenes as limiting reagents, requiring only stoichiometric amounts of EDA. The protocol is performed in one-pot fashion without the need of slow addition of EDA. The diastereoselectivity of the current system can be tuned by using different porphyrin ligands or additives, giving either trans- or cis-dominant cyclopropanes. The asymmetric cyclopropanation was also demonstrated with the use of chiral cobalt porphyrin complexes. PMID- 14535802 TI - Synthesis of orthogonally protected lanthionines. AB - Synthetic approaches to the lantibiotics, a family of thioether-bridged antimicrobial peptides, require flexible synthetic routes to a variety of orthogonally protected derivatives of lanthionine 1. The most direct approaches to lanthionine involve the reaction of cysteine with an alanyl beta-cation equivalent. Several possibilities exist for the alanyl beta-cation equivalent, including direct activation of serine under Mitsunobu conditions: however, the low reactivity of sulfur nucleophiles in the Mitsunobu reaction has previously precluded its use in the synthesis of the lantibiotics. We report here a new approach to the synthesis of protected lanthionine, using a novel variant of the Mitsunobu reaction in which catalytic zinc tartrate is used to enhance the nucleophilicity of the thiol. In the course of these studies, we have also demonstrated that the synthesis of lanthionine from trityl-protected beta iodoalanines is prone to rearrangement, via an aziridine, to give predominantly trityl-protected alpha-iodo-beta-alanines, and hence norlanthionines, as the major products. PMID- 14535803 TI - Synthesis of a cyclic peptide containing norlanthionine: effect of the thioether bridge on peptide conformation. AB - Two diastereomeric analogues of ring C of nisin incorporating a novel norlanthionine residue have been synthesized via a triply orthogonal protecting group strategy. A full structural study was carried out by NMR, which elucidated the conformational properties of the two peptides and enabled the identity of each diastereoisomer to be proposed. PMID- 14535804 TI - Synthesis of cyclic hexameric porphyrin arrays. Anchors for surface immobilization and columnar self-assembly. AB - To investigate new architectures for the self-assembly of multiporphyrin arrays, a one-flask synthesis of a shape-persistent cyclic hexameric array of porphyrins was exploited to prepare six derivatives bearing diverse pendant groups. The new arrays contain 6-12 carboxylic acid groups, 12 amidino groups, 6 thiol groups, or 6 thiol groups and 6 carboxylic acid groups in protected form (S-acetylthio, TMS ethyl, TMS-ethoxycarbonyl). The arrays contain alternating Zn and free base (Fb) porphyrins or all Zn porphyrins. The one-flask synthesis entails a template directed, Pd-mediated coupling of a p/p'-substituted diethynyl Zn porphyrin and a m/m'-substituted diiodo Fb porphyrin. The porphyrin building blocks (trans A(2)B(2), trans-AB(2)C) contain the protected pendant groups at nonlinking meso positions. A self-assembled monolayer (SAM) of a Zn(3)Fb(3) cyclic hexamer containing one thiol group on each porphyrin was prepared on a gold electrode and the surface-immobilized architecture was examined electrochemically. Together, the work reported herein provides cyclic hexameric porphyrin arrays for studies of self-assembly in solution or on surfaces. PMID- 14535805 TI - The nicotinic pharmacophore: thermodynamics of the hydrogen-bonding complexation of nicotine, nornicotine, and models. AB - The thermodynamics of the hydrogen-bonding complexation of the acetylcholine agonists nicotine and nornicotine and of model pyridines, pyrrolidines, and N methylpyrrolidines has been measured in CCl(4) by FTIR spectrometry toward a reference hydrogen-bond donor, 4-fluorophenol. Various methods are devised for measuring separately the hydrogen-bond acceptor strength of each nitrogen of nicotine and nornicotine: variation of the stoichiometry of complexation; correlations with electrostatic potentials on nitrogens and with substituent constants in the series of 3-substituted pyridines, 2-substituted pyrrolidines, and 2-substituted N-methylpyrrolidines; and linear free energy relationships between 4-fluorophenol and hydrogen fluoride hydrogen-bonded complexes. It is consistently found that nicotine and nornicotine have two active hydrogen-bond acceptor sites, the pyridine and pyrrolidine nitrogens, and that ca. 90% (for nicotine) and 80% (for nornicotine) of the 1:1 hydrogen-bonded complexes are formed to the pyridine nitrogen, although the pyrrolidine nitrogen is the first protonation site of nicotine and nornicotine in water. The low hydrogen-bond basicity of the pyrrolidine nitrogen in nicotine is mainly explained by the inductive electron-withdrawing and steric effects of the 2-(3-pyridyl) substituent. The partition of the Gibbs energy of the isomerism of complexation (AH...Nsp(2) <==> AH...Nsp(3)) into enthalpic and entropic contributions shows that the selectivity in favor of the pyridine nitrogen is driven by entropy. It is important to recognize the bifunctionality of nicotine in hydrogen bonding for understanding its lipophilicity and molecular recognition in non protonic media. When monoprotonated on their sp(3) nitrogen, nicotine and nornicotine keep, through their sp(2) nitrogen, a significant hydrogen-bond basicity which is greater than that of the ester group of acetylcholine. PMID- 14535806 TI - Chiral hydroperoxides as oxygen source in the catalytic stereoselective epoxidation of allylic alcohols by sandwich-type polyoxometalates: control of enantioselectivity through a metal-coordinated template. AB - The epoxidation of allylic alcohols is shown to be efficiently and selectively catalyzed by the oxidatively resistant sandwich-type polyoxometalates, POMs, namely [WZnM(2)(ZnW(9)O(34))(2)](q)(-) [M = OV(IV), Mn(II), Ru(III), Fe(III), Pd(II), Pt(II), Zn(II); q = 10-12], with organic hydroperoxides as oxygen source. Conspicuous is the fact that the nature of the transition metal M in the central ring of polyoxometalate affects significantly the reactivity, chemoselectivity, regioselectivity, and stereoselectivity of the allylic alcohol epoxidation. For the first time, it is demonstrated that the oxovanadium(IV)-substituted POM, namely [ZnW(VO)(2)(ZnW(9)O(34))(2)](12-), is a highly chemoselective, regioselective, and also stereoselective catalyst for the clean epoxidation of allylic alcohols. A high enantioselectivity (er values up to 95:5) has been achieved with [ZnW(VO)(2)(ZnW(9)O(34))(2)](12)(-) and the sterically demanding TADOOL-derived hydroperoxide TADOOH as regenerative chiral oxygen source. Thus, a POM-catalyzed asymmetric epoxidation of excellent catalytic efficiency (up to 42 000 TON) has been made available for the development of sustainable oxidation processes. The high reactivity and selectivity of this unprecedented oxygen transfer process are mechanistically rationalized in terms of a peroxy-type vanadium(V) template. PMID- 14535807 TI - Calculations of substituent and solvent effects on the kinetic isotope effects of Menshutkin reactions. AB - Nitrogen, deuterium, halogen, and carbon kinetic isotope effects have been modeled for the Menshutkin reaction between methyl halides and substituted N,N dimethylaniline at the HF/6-31G(d) level of theory augmented by the C-PCM continuum solvent model for several solvents. Systematic changes in geometries of the transition states and Gibbs free energies of activation have been found with phenyl ring substituents, solvent, and the leaving group. Kinetic isotope effects also change systematically; however, these changes are predicted to be small, inside the usual precision of the experimental measurements. On the contrary, no correlation has been found between the kinetic isotope effects and the Hammett constants for para substituents. Thus opposite to previous assumptions, our results indicate that kinetic isotope effects on the Menshutkin reaction cannot be used to predict the position of the transition state on the reaction coordinate. PMID- 14535808 TI - Anomalous C-C bond cleavage in sulfur-centered cation radicals containing a vicinal hydroxy group. AB - 1,3-dithianyl cation radicals having alpha-hydroxy-neopentyl or similar groups in position 2, which are generated via oxidative photoinduced electron transfer, undergo anomalous fragmentation necessitating refinement of the accepted mechanism. Experimental and computational data support a rationale in which proton abstraction from the hydroxy group in the initial cation radical does not cause a Grob-like fragmentation, but rather produces a neutral radical species, the alkoxy radical, that undergoes fragmentation in either direction, i.e., cleaving the C-C bond to dithiane or to the tertiary alkyl group. PMID- 14535809 TI - A redox-mediated molecular brake: dynamic NMR study of 2-[2 (methylthio)phenyl]isoindolin-1-one and S-oxidized counterparts. AB - A redox-mediated molecular brake based on the sulfide-sulfoxide redox cycle is illustrated by modulation of the rotation rate of an N-Ar "shaft" by varying the oxidation state of sulfur in 2-[2-(sulfur-substituted)phenyl]isoindolin-1-ones. N Ar rotational barriers in methylsulfinyl (2) and methylsulfonyl (3) derivatives (13.6 kcal mol(-1)) are approximately 5 kcal mol(-1) higher than sulfide 1. Rate reduction for N-Ar rotation is approximately 10(4) s(-1) (280 K) upon oxidation. Correlated N-pyramidalization/N-Ar rotation reduces the effectiveness of the brake by decreasing the energy barrier to N-Ar bond rotation. PMID- 14535810 TI - Ferromagnetic behavior of formyl-group-carrying stable thioaminyl radicals. AB - Four formyl-group-carrying thioaminyl radicals were generated, and one radical could be isolated as radical crystals. Magnetic susceptibility measurements of the isolated radical showed a ferromagnetic regular linear-chain interaction of 2J/k(B) = 3.2 K, which was explained in terms of the X-ray crystallographic results. PMID- 14535811 TI - Zn(II)-catalyzed thiolysis of oxiranes in water under neutral conditions. AB - Thiolysis of a variety of 1,2-epoxides in water at 30 degrees C and pH 7.0 is strongly accelerated by ZnCl(2) (10 mol %) except when amino- and carboxythiophenol are used. The aqueous medium and the catalyst were recovered and reused in various runs without affecting the efficiency of the process. PMID- 14535812 TI - Short synthesis of enantiopure C2-symmetric 1,2:4,5-diepoxypentane and "pseudo" C2-symmetric 3-azido-1,2:4,5-diepoxypentane from arabitol. AB - On the basis of our previously described selective protection of arabitol as its 1,2:4,5-bis-pentylidene acetal 5, we report a straightforward synthesis of the novel "pseudo"-C(2)-symmetric 3-azido-1,2:4,5-diepoxypentane building block 4 in 6 steps from arabitol. Using a similar synthetic route, an improved synthesis of the C(2)-symmetrical 1,2:4,5-bis-epoxypentane building block 1 is described, also in 6 steps from arabitol. Both enantiomers of 1 and 4 are accessible, and all reactions involved are easily amenable for large-scale synthesis. PMID- 14535813 TI - 2-azabicyclo[2.2.2]octa-3,5-dione via a nitrile Diels-Alder reaction. AB - The hetero-Diels-Alder reaction of an electron-deficient nitrile, p toluenesulfonyl cyanide, with the silyl enol ether of cyclohexenone produced a hydrolytically sensitive [4 + 2] adduct in good yield. Use of Mander's reagent, ethyl cyanoformate, with the same diene, produced an unstable adduct. Hydrolysis of the tosyl cyanide adduct resulted in the isoquinuclidinone 2-aza[2.2.2]octa 3,5-dione as a crystalline compound in excellent yield from the nitrile. PMID- 14535814 TI - Photoaddition reactions of acetylene and butadiyne derivatives to benzodithiophene. AB - The photochemical [2pi +2pi] cycloaddition of dimethyl acetylenedicarboxylate to benzo[1,2-b:4,5-b']dithiophene has been used to synthesize substituted cyclobuta[b]thieno[2,3-f][1]benzothiophene. The first [2pi + 2pi] photocycloaddition reaction of a series of butadiynes to benzodithiophene is reported to yield regioselective and acetylene-substituted cyclobutene derivatives containing an aromatic thiophene moiety. PMID- 14535815 TI - Use of sonication for the coupling of sterically hindered substrates in the phenolic Mitsunobu reaction. AB - A vast rate increase in the Mitsunobu reaction of phenols with alcohols where either or both are sterically hindered has been achieved by the use of high concentration combined with sonication. PMID- 14535816 TI - Syntheses of regioisomerically pure 5- or 6-halogenated fluoresceins. AB - Three routes to regioisomerically pure 5- and 6-iodofluoresceins or 5- and 6 bromofluoresceins are described. The first, shown in Scheme 1, involves diazotization/iodination of the corresponding aminofluoresceins. In the second approach (Scheme 2) a mixture of regioisomeric fluoresceins was prepared, and the 5-bromo isomers were isolated as the ring closed diacetates 9b and 11 by fractional crystallization. Scheme 3 shows an approach to sulfonic acid derivatives 3 and 4 of 5-iodofluorescein. This is the most convenient procedure of the three, and it is particularly useful as sulfofluoresceins have more favorable water solubility characteristics than fluoresceins that lack the sulfonic acid group. PMID- 14535817 TI - A facile asymmetric route to (-)-aphanorphine. AB - 8O-methylaphanorphine was synthesized from 4-methoxyphenylacetaldehyde in 36% overall yield and in nine steps, featuring the formation of ring B via a Friedel Crafts alkylative cyclization with the concomitant stereospecific introduction of the benzylic quaternary carbon center. The current work constitutes an efficient enantioselective formal synthesis of 3-benzazepine marine alkaloid (-) aphanorphine. PMID- 14535818 TI - Oxidative aromatization of 9,10-dihydroanthracenes using molecular oxygen promoted by activated carbon. AB - Substituted 9,10-dihydroanthracenes were oxidatively aromatized to the corresponding anthracenes effectively by using molecular oxygen as an oxidant and activated carbon (Darco KB, Aldrich, Inc.) as a promoter in xylene. PMID- 14535819 TI - Facile one-pot synthesis of aromatic and heteroaromatic sulfonamides. AB - A series of arene and heteroarene sulfonamides were prepared in one vessel from aryl and heteroaryl bromides via conversion into the corresponding Grignard reagents using either magnesium or isopropylmagnesium chloride and subsequent reaction with sulfur dioxide, sulfuryl chloride, and an amine. PMID- 14535820 TI - Highly enantioselective conjugate addition of diethylzinc to acyclic enones with fine-tunable phosphite-pyridine ligands. AB - A new series of fine-tunable phosphite-pyridine (P,N) ligands derived from (S)-2 amino-2'-hydroxy-6,6'-dimethyl-1,1'-biphenyl and (S)-2-amino-2'-hydroxy-4,4',6,6' tetramethyl-1,1'-biphenyl was employed in Cu(I)-catalyzed conjugate addition of diethylzinc to acyclic enones. Excellent enantioselectivities (up to 98% ee) and highly catalytic activities were achieved for a variety of acyclic enones. PMID- 14535821 TI - Using cyanide to put noble gases inside C60. AB - After fullerenes are heated in the presence of a noble gas or an unreactive molecule at 650 degrees C and 3000 atm pressure, a small fraction of the fullerene molecules contain the atom or molecule. The incorporation fraction is greatly enhanced by adding potassium cyanide to the reaction mixture. The details of the preparation are described here. PMID- 14535822 TI - Evidence for 5-center 4-electron bonding in (C...H...C...H...C) array. AB - This paper brings new independent evidence for 5-center 4-electron bonding, whose existence in (C...H...C...H...C) fragment of the molecule I was anticipated in the recent study by Tantillo and Hoffmann. The evidence is based on the applications of the so-called multicenter bond indices, recently proposed as new efficient tool for the detection and localization of multicenter bonding in molecules. PMID- 14535823 TI - A novel method for the preparation of alpha,alpha'-difluoroesters and acids using BrF3. AB - Alkyl-, haloalkyl-, and ketoalkyl-2-ethoxycarbonyl-1,3-dithianes were easily made from the appropriate primary or secondary alkyl bromides, 1,3-dithiane, and ethyl chloroformate. They were reacted with BrF(3) to form the corresponding alpha,alpha-difluoro esters in 65-75% yield. Reaction conditions are very mild (1 2 min, 0 degrees C). The two sulfur atoms of the dithiane are essential for the reaction. PMID- 14535825 TI - Immature lung and acute lung injury. AB - Acute lung injury occurs mostly in the very low birth weight and extremely low birth weight infants. The pathological process leading to acute lung injury includes immature and/or diseased lung that experienced oxidative stress, inflammation and mechanical insult with the bronchial, alveolar and capillary injuries and cell death. It may be the first step to the subsequent development of chronic lung disease of prematurity or bronchopulmonary dysplasia. The mechanisms of lung injury are extensively investigated in the experimental models and clinical studies, mostly performed on the adult patients. At present, the explanations of the mechanism(s) leading to lung tissue injury in tiny premature babies are just derived from these studies. Acute lung injury seems to be rather a syndrome than a well-defined nosological unit and is of multifactorial etiology. The purpose of this review is to discuss the main factors contributing to the development of acute lung injury in the very low or extremely low birth weight infants--lung immaturity, mechanical injury, oxidative stress and inflammation. Nevertheless, numerous other factors may influence the status of immature lung after delivery. PMID- 14535826 TI - Short-term dynamics of coherence between respiratory movements, heart rate, and arterial pressure fluctuations in severe acute brain disorders. AB - In our previous study, healthy volunteers showed considerable short-term dynamics and patterns of the coherence of high time resolution between respiratory movements (RESP), heart rate fluctuations (HRF), and arterial blood pressure fluctuations (BPF). These are physiological indicators of autonomic short-term coordination mediated mainly by the brainstem which could be impaired in severe brain disorders. We hypothesized a direct or indirect impairment of these functions by these disorders and examined these patterns in 16 patients suffering from severe brain disorders. We calculated partial and ordinary coherence sequences and found almost the same patterns of coherence sequences as in healthy volunteers, but a distinctly reduced frequency of pattern incidence in patients (2.8+/-1.5/10 min/patient and 9.5+/-2.8/10 min/subject, P<0.05). Furthermore, there is a significantly smaller frequency of HRF-related patterns in patients with poor outcome, compared with those in patients with good outcome (1.8+/ 0.8/10 min/patient and 4.5+/-2.7/10 min/patient, P<0.05). We conclude that severe brain disorders reduce physiological short-term dynamics of autonomic coordination patterns in the mean values of patients, but not in every patient. PMID- 14535827 TI - Remodeling of the rat saphenous vein network in response to long-term gravitational load. AB - Our main objective was to test whether chronic orthostatic body position induces network changes in the saphenous vein superficial tributary system of the rat. Fourteen male Sprague-Dawley rats were kept in tilted tube cages (45-degree head up position) for two weeks to induce chronic gravitational load to their leg veins. Ten animals housed in normal cages and four animals kept in horizontally positioned tube cages served as controls. The whole superficial network of the left saphenous vein was microprepared surgically under anesthesia, superfused with saline and observed under a videomicroscope, while normal flow and pressure were maintained in the lumen. Branching angles, lengths of venous segments and their diameters were measured offline from digitized images using special image analyzing software. Several branching angles at the popliteal confluence were significantly reduced by 12.5-15.8 %. The in vivo diameter of the main branch (936+/-34 vs. 805+/-44 microm) and of one of the popliteal tributaries (776+/-38 vs. 635+/-36 microm) increased (p<0.05), comparing vessels from tilted animals with those from normal controls. Maintaining the animals in horizontal tube cages did not induce the above alterations. The increased diameters and reduced branching angles of the saphenous vein network observed are adaptive responses of the venous network to a long-term gravitational load. PMID- 14535828 TI - Influence of different oxygen modes on the blood oxygen transport and prooxidant antioxidant status during hepatic ischemia/reperfusion. AB - Oxygen supply was corrected in rabbits during the hepatic ischemia/reperfusion by means of different breathing mixtures: hypoxic (14.8 % O(2)+85.2 % N(2)), hyperoxic (78 % O(2)+20.2 % N(2)+ 1.8 % CO(2)), or hypercapnic (5 % CO(2) in air). Hepatic ischemia was induced for 30 min by ligation of hepatic artery, reperfusion period lasted 120 min. Indices of blood oxygen transport (p50(act), pCO(2), pH, pO(2), etc.) and prooxidant-antioxidant balance (Schiff bases, conjugated dienes, catalase, retinol, alpha-tocopherol) were measured in the blood and liver. The severity of reperfusion damage was evaluated by the activities of alanine and aspartate aminotransferases (ALT, AST) in the blood. Hepatic ischemia/reperfusion resulted in higher p50(act) in hepatic venous and mixed venous blood in all experimental groups. The changes of p50(act) were most marked in the hypercapnic group and were the weakest in the hypoxic group. The rise in p50(act) was accompanied by higher levels of lipid peroxidation products, ALT and AST in blood and liver homogenates, and by a simultaneous fall of alpha tocopherol and retinol concentrations, except in the hypoxic group. Catalase activity at the end of reperfusion increased under normoxia, decreased under hyperoxia or hypercapnia and did not change under hypoxia. The moderate hypoxia during reperfusion was accompanied by a better balance between the mechanisms of reactive oxygen species production and inactivation that may be observed by optimal changes in p50act and reduced the hepatic damage in this pathological condition. PMID- 14535829 TI - Inhibitory effects of Bcl-2 on mitochondrial respiration. AB - In contrast to the well-established anti-apoptotic effect of Bcl-2 protein, we have recently demonstrated that Bcl-2 overexpression by vaccinia virus causes apoptosis in BSC-40 cells, while it prevents apoptosis in HeLa G cells. Given the key role of mitochondria in the process of apoptosis, we focused on effects of Bcl-2 expression on mitochondrial energetics of these two cell lines. In this study we present data indicating that BSC-40 cells derive their ATP mainly from oxidative phosphorylation whereas HeLa G cells from glycolysis. More importantly, we show that in both cell lines, Bcl-2 inhibits mitochondrial respiration and causes a decrease of the ATP/ADP ratio. However, it appears that BSC-40 cells cannot sustain this decrease and die, while HeLa G cells survive, being adapted to the low ratio of ATP/ADP maintained by glycolysis. Based on this observation, we propose that the outcome of Bcl-2 expression is determined by the type of cellular ATP synthesis, namely that Bcl-2 causes apoptosis in cells relying on oxidative phosphorylation. PMID- 14535830 TI - Effect of resveratrol on some activities of isolated and in whole blood human neutrophils. AB - Resveratrol, which is a polyphenol present in red wines and vegetables included in human diets, exerts many biological effects. The aim of the present study was to investigate its effect on some activities of polymorphonuclear leukocytes, particularly the generation of superoxide anion ((O2)(-)) in whole blood, hypochlorous acid (HOCl) and nitric oxide (NO) production by isolated cells, and chemotaxis. Resveratrol showed significant dose-dependent inhibitory effect on all these activities. In particular, it inhibited O2(-) generation in stimulated but not in resting neutrophils, decreased HOCl much more than O2(-) production indicating an effect on myeloperoxidase secretion since HOCl production is directly and proportionally dependent on O2(-) generation and reduced cell motility. The small dose of resveratrol (4.38 nM) used is attainable with a diet including red wine and vegetables confirming its protective role against some pathological processes such as inflammation, coronary heart disease, and cancer. PMID- 14535831 TI - Protective influence of vitamin E on antioxidant defense system in the blood of rats treated with cadmium. AB - The effects of acute exposure to cadmium (Cd) on the blood antioxidant defense system, lipid peroxide concentration and hematological parameters, as well as the possible protective role of vitamin E were studied. Male Wistar albino rats (3 months old) were treated with cadmium (0.4 mg Cd/kg b.m., i.p., 24 h before the experiment) or with vitamin E + Cd (20 IU Vit E/kg b.m., i.m., 48 h + 0.4 mg Cd/kg b.m., i.p., 24 h before the experiment). The hematological parameters were assessed: red blood cell counts, hematocrit value and hemoglobin concentration were significantly decreased in the blood of Cd-treated rats. Intoxication with cadmium was also followed by significantly increased lipid peroxide concentrations. We also observed increased activity of antioxidant defense enzymes: copper zinc containing superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and glutathione-S-transferase as well as concentrations of non-enzymatic components of antioxidant defense system: reduced glutathione, vitamin C and vitamin E. Pretreatment with vitamin E exhibited a protective role on the toxic effects of cadmium on the hematological values, lipid peroxide concentration as well as on enzymatic and non-enzymatic components of antioxidant defense system. PMID- 14535832 TI - Kinetics of cardiac RyR channel gating studied at high temporal resolution. AB - Measurements of ryanodine receptor (RyR) activity during dynamic changes of calcium concentration have suggested that RyR has at least four calcium binding sites, and that activation transpires as an increase in the activity within the high open probability H-mode. Binding of several Ca2+ ions within the H-mode should manifest itself in the steady-state RyR activity by the presence of multiple closed times. However, previously only two closed times were detected in the H-mode of RyR activity. Here we recorded steady-state activity of single cardiac RyRs with high temporal resolution and compared it to data simulated under the same conditions using our previously published model of RyR gating. At a 10 kHz resolution, the closed time histograms of both experimental and simulated data had three exponential components. The closed times of simulated data were not significantly different from those obtained experimentally. After filtering at 2 kHz, only two exponential closed time components with time constants not significantly different from those previously published could be detected in both experimental and simulated records. The conformity of the steady state experimental data to the model derived from the dynamic data provides further support for the idea that RyRs need binding of multiple Ca2+ ions to open. PMID- 14535833 TI - Frequency-domain lifetime fluorometry of double-labeled creatine kinase. AB - Myofibril-bound creatine kinase EC 2.7.3.2 (CK), a key enzyme of muscle energy metabolism, has been selected for studies of conformational changes that underlie the cellular control of enzyme activity. For fluorescence spectroscopy measurements, the CK molecule was double-labeled with IAF (5 iodoacetamidofluorescein) and ErITC (erythrosin 5'-isothiocyanate). Measurement of fluorescence resonance energy transfer (FRET) from fluorescein to erythrosin was used to obtain information about the donor-acceptor pair distance. Frequency domain lifetime measurements evaluate the donor-acceptor distance in the native CK molecule as 7.8 nm. The Forster radius equals 5.3 nm with the resolution range from 0.2 to 1.0 nm. Erythrosin-fluorescein labeling (EFL) was tested for artificial conformational changes of the CK molecule with high-salt concentration treatment. The transition distance, defined by His-97 and Cys-283 and derived from a 3D model equals 0.766 nm for the open (inactive) form and 0.277 nm for the closed (reactive) form of the CK molecule. In this way, the resolution range of the used spectroscopy method is significant, concerning the difference of 0.489 nm. Nevertheless, the CK enzyme activity, assessed by the hexokinase-coupled assay, was diminished down to 1 % of the activity of the native enzyme. EFL is suitable for description of conformational behavior implied from the regulation of creatine kinase. However, the observed inhibition restricts EFL to studies of conformational changes during natural catalytic activity. PMID- 14535834 TI - Chloride cotransport in the membrane of earthworm body wall muscles. AB - The resting membrane potential (V(m)) of isolated somatic longitudinal muscles of the earthworm Lumbricus terrestris was studied by glass microelectrodes. The inhibition of chloride permeability by low pH did not affect V(m) of the muscle fibers in isolated somatic longitudinal muscles of the earthworm Lumbricus terrestris which was -48.7 mV (inside negative) at pH 7.3 and -49.1 at pH 5.6. On the other hand, bathing the muscles in Cl(-) and Na(+)-free solutions, or application of the chloride transporter inhibitor furosemide and Na(+)-K(+) ATPase inhibitor ouabain depolarized the V(m) by 3-5 mV. The effects of a Cl(-) free solution and ouabain were not additive. This demonstrates relatively small contribution of equilibrium potential for Cl(-) to the resting membrane potential and electrogenic effect of Na(+)K(+)-ATPase which is dependent on the supply of Na(+)(i) ions by furosemide-sensitive and Cl(-)(e)- and Na(+)(e)-dependent electroneutral transport (most probably Na(+)K(+)Cl(-) cotransport). PMID- 14535835 TI - Dynamics of cytokine production in human peripheral blood mononuclear cells stimulated by LPS or infected by Borrelia. AB - To specify the role of individual cytokines in the immune response to pyrogens, isolated and cultivated human peripheral blood mononuclear cells (PBMC) were used for the experiments. Different pyrogens (lipopolysaccharide from Escherichia coli - LPS and live Borrelia afzelii) were applied and the time course of changes in concentrations of different cytokines in the medium was followed using the ELISA method. It was found that nonstimulated human PBMC proliferate under in vitro conditions and produce IL-6, TNF-alpha, IL-10 and finally also IL-1beta. Productions of IL-12 and INF-gamma are not changed. Proliferation of PBMC is potentiated after incubation with LPS or live Borrelia. PBMC stimulated by LPS increase the net production (stimulated minus unstimulated) of IL-1beta and TNF alpha significantly, while production of IL-6 was smaller. A delayed increase in the production of IL-10 was also observed. Productions of IL-12 and INF-gamma were not influenced. In contrast to LPS, stimulation of PBMC with live Borrelia, increases also the production of IL-12 and IFN-gamma, besides IL-1beta, TNF alpha, IL-6 and IL-10. Productions of IL-1beta, IL-6 and TNFalpha increased immediately after incubation with both LPS and Borrelia, while productions of IL 12 and INF-gamma begin to increase 8 hours and production of IL-10 12 hours after stimulation. Data indicate that stimulation with different pyrogens may activate the cells of the immune cascade in a different way. Stimulation of BPMC by LPS seems to activate the initial steps of the immune response (macrophages and granulocytes) only, while infection with live Borrelia also stimulates the later phase of the immune response, probably due to effect of initially produced cytokines. PMID- 14535836 TI - Modulation of ionizing radiation-induced apoptosis and cell cycle arrest by all trans retinoic acid in promyelocytic leukemia cells (HL-60). AB - Acute promyelocytic leukemia is characterized by a block of myeloid differentiation. The incubation of cells with 1 micromol/l all-trans retinoic acid (ATRA) for 72 h induced differentiation of HL-60 cells and increased the number of CD11b positive cells. Morphological and functional changes were accompanied by a loss of proliferative capacity. Differentiation caused by preincubation of leukemic cells HL-60 with ATRA is accompanied by loss of clonogenicity (control cells: 870 colonies/10(3) cells, cells preincubated with ATRA: 150 colonies/10(3) cells). D0 for undifferentiated cells was 2.35 Gy, for ATRA differentiated cells 2.46 Gy. Statistical comparison of clonogenity curves indicated that in the whole range 0.5-10 Gy the curves are not significantly different, however, in the range 0.5-3 Gy ATRA differentiated cells were significantly more radioresistant than non-differentiated cells. When HL-60 cells preincubated with 1 micromol/l ATRA were irradiated by a sublethal dose of 6 Gy, more marked increase of apoptotic cells number was observed 24 h after irradiation and the surviving cells were mainly in the G1 phase of the cell cycle, while only irradiated cells were accumulated in G(2) phase. Our results imply that preincubation of cells with ATRA accelerates apoptosis occurrence (24 h) after irradiation by high sublethal dose of 6 Gy. Forty-eight hours after 6 Gy irradiation, late apoptotic cells were found in the group of ATRA pretreated cells, as determined by APO2.7 positivity. This test showed an increased effect (considering cell death induction) in comparison to ATRA or irradiation itself. PMID- 14535837 TI - Altered blood-brain barrier permeability and its effect on the distribution of Evans blue and sodium fluorescein in the rat brain applied by intracarotid injection. AB - The aim was to study the blood-brain permeability according to the distribution in the rat brain of Evans blue (EB) and sodium fluorescein (NaFl) administered by an intracarotid injection. Eighteen animals were divided into six groups according to the state of the blood-brain barrier (BBB) at the moment when the dyes were being applied. In the first two groups, the BBB was intact, in groups 3 and 4 the barrier had been opened osmotically prior to the application of the dyes, and in groups 5 and 6 a cellular edema was induced by hyperhydration before administration of the dyes. The intracellular and extracellular distribution of the dyes was studied by fluorescence microscopy. The histological picture thus represented the morphological correlate of the way BBB permeability had been changed before the application of the dyes. PMID- 14535838 TI - Changes in neuronal activity of the inferior colliculus in rat after temporal inactivation of the auditory cortex. AB - The role of cortico-tectal pathways in auditory signal processing was studied in anesthetized rats by comparing the extracellular single unit activity in the inferior colliculus (IC) before and after functional ablation of the auditory cortex (AC) by tetrodotoxin (TTX). The responses of several IC neurons to sound stimuli were simultaneously recorded with a 16-channel electrode probe introduced into the IC. Click-evoked middle latency responses (MLR) recorded from the AC were suppressed for several hours after TTX injection. During AC inactivation the firing rate of IC neurons increased (40 % of neurons), decreased (44 %) or did not change (16 %) in comparison with control conditions. In several IC neurons, TTX injection resulted in alterations in the shape of the rate-level functions. Response thresholds, tuning properties and the type of discharge pattern of IC neurons were not altered during AC inactivation. However, in one-third of the neurons, the initial part of the response was less altered than the later, sustained part. In two-thirds of neuronal pairs, functional decortication resulted in a change in the cross-correlation coefficient. The results reveal the complex changes that appear in IC neuronal activity after functional ablation of the ipsilateral auditory cortex. PMID- 14535839 TI - Subcellular targets of mercaptoborate (BSH), a carrier of 10B for neutron capture therapy (BNCT) of brain tumors. AB - The transformed C6 glial cells in cultures were treated with sodium mercaptoborate (Na(2)B(12)H(11)SH, BSH), a carrier of atomic targets ((10)B) of thermal neutrons for the neutron capture therapy of brain tumors. As shown by light microscopy, the therapeutic dose of BSH (100 microg/ml) did not alter the gross morphology and growth of the population of cells within a 72 h treatment interval. Electron microscopic analysis of these cells revealed activation of nucleoli and, occasionally, enlarged and bifurcated mitochondria. After 200 microg BSH/ml and 72 h treatment, growth of the cell population was inhibited and ultrastructural changes became more profound. They included condensation of chromatin and its allocation to the nuclear envelope which formed deeper invaginations. Mitochondria further increased in size and were characterized by slim or angular cristae. Moreover, in circumscribed segments of some of the slightly swollen mitochondria their cristae disappeared or were reduced to fine pouch-like structures localized near the continuous outer membrane, suggestive for a non-destructive restructuring of the inner mitochondrial membrane. The smooth pinocytotic vesicles near the plasma membrane, lysosomes and heterogeneous dense bodies were more frequent. The revealed subcellular targets of BSH may initiate the development of pharmacological protocols aimed to further improve the tolerance to BSH by the healthy tissues of patients undergoing BNCT of brain tumors, e.g. by intervention into the oxidative stress triggered likely by the altered mitochondria. PMID- 14535840 TI - Effects of lower body negative pressure on cardiac and vascular responses to carotid baroreflex stimulation. AB - The aim of this study was to assess carotid baroreflex responses during graded lower body negative pressure (LBNP). In 12 healthy subjects (age 29+/-4 years) we applied sinusoidal neck suction (0 to -30 mmHg) at 0.1 Hz to examine the sympathetic modulation of the heart and blood vessels and at 0.2 Hz to assess the effect of parasympathetic stimulation on the heart. Responses to neck suction were determined as the change in spectral power of RR-interval and blood pressure from baseline values. Measurements were carried out during progressive applications (0 to -50 mmHg) of LBNP. Responses to 0.1 and 0.2 Hz carotid baroreceptor stimulations during low levels of LBNP (-10 mmHg) were not significantly different from those measured during baseline. At higher levels of LBNP, blood pressure responses to 0.1 Hz neck suction were significantly enhanced, but with no significant change in the RR-interval response. LBNP at all levels had no effect on the RR-interval response to 0.2 Hz neck suction. The unchanged responses of RR-interval and blood pressure to neck suction during low level LBNP at -10 mmHg suggest no effect of cardiopulmonary receptor unloading on the carotid arterial baroreflex, since this LBNP level is considered to stimulate cardiopulmonary but not arterial baroreflexes. Enhanced blood pressure responses to neck suction during higher levels of LBNP are not necessarily the result of a reflex interaction but may serve to protect the circulation from fluctuations in blood pressure while standing. PMID- 14535841 TI - Familial dysbetalipoproteinemia in three patients with apoE 2*(Arg136-->Cys) gene variant. AB - Apolipoprotein E (apoE) is a polymorphic protein which occurs in three common isoforms and more than 25 rare variants. Some of the rare apoE variants have been implicated in a dominant mode of inheritance of familial dysbetalipoproteinemia (FD). We have identified three unrelated apoE 2*(Arg136-->Cys) carriers with FD. This finding supports the notion that although apoE 2*(Arg136-->Cys) mutation is perhaps not sufficient to cause FD itself, the presence of other genetic and/or environmental factors can lead to the phenotypic expression of the disease in the carriers. PMID- 14535842 TI - Action of GABA-B antagonist on cortical epileptic afterdischarges in rats is similar to that of GABA-A antagonist. AB - Threshold intensities for epileptic phenomena induced by cortical stimulation were used for comparison of the action of GABA-B and GABA-A antagonists in rats with implanted electrodes. Both CGP 35348 (200 mg/kg i.p.) and bicuculline (4 mg/kg i.p.) significantly decreased thresholds for spike-and-wave afterdischarges and their motor counterpart (clonic seizures) whilst transition into the second, limbic type of afterdischarge as well as threshold for movements directly bound to stimulation remained uninfluenced by either drug. PMID- 14535843 TI - Engineering conduits to resemble natural vascular tissue. AB - Research on the use of biomaterials as vascular substitutes has been dominated in the past by studies focusing on the properties of the materials used. These need to be resistant to protein absorption and should allow cell colonization and integration of the biomaterial within recipient tissue. Despite promising perspectives related both to the appearance of new synthetic materials with biomedical applications and to developments in arterial replacement techniques, there are still many unresolved problems. These include the appearance of occlusive phenomena when using medium- or small-calibre arterial substitutes, and the lack of growth and adaptability of the biomaterial to changes in the host. Notwithstanding, the tissue-engineering techniques developed in recent decades have vastly improved prosthetic behaviour at the blood/biomaterial interface. There is also no doubt that developments in engineered arterial grafts made of natural biomaterials colonized with cells harvested from the recipient patient are drawing us closer and closer to achieving the living graft. PMID- 14535844 TI - Tyr-298 in ephrinB1 is critical for an interaction with the Grb4 adaptor protein. AB - The Eph family of receptor tyrosine kinases and their membrane-bound ligands, the ephrins, are thought to play a role in the regulation of cell adhesion and migration during development by mediating cell-to-cell signalling events. The transmembrane ephrinB protein is a bidirectional signalling molecule that sends a forward signal through the activation of its cognate receptor tyrosine kinase residing on another cell. The reverse signal is transduced into the ephrinB expressing cell via tyrosine phosphorylation of its conserved C-terminal cytoplasmic domain. Previous work from our laboratory has implicated the activated FGFR1 (fibroblast growth factor receptor 1) as a regulator of a de adhesion signal that results from overexpression of ephrinB1. In the present study, we report the isolation of Xenopus Grb4 (growth-factor-receptor-bound protein 4), an ephrinB1-interacting protein, and we show that when expressed in Xenopus oocytes, ephrinB1 interacts with Grb4 in the presence of an activated FGFR1. Amino acid substitutions were generated in Grb4, and the resulting mutants were expressed along with ephrinB1 and an activated FGFR in Xenopus oocytes. Co immunoprecipitation analysis shows that the FLVR motif within the Src homology 2 domain of Xenopus Grb4 is vital for this phosphorylation-dependent interaction with ephrinB1. More importantly, using deletion and substitution analysis we identify the tyrosine residue at position 298 of ephrinB1 as being required for the physical interaction with Grb4, whereas Tyr-305 and Tyr-310 are dispensable. Moreover, we show that the region between amino acids 301 and 304 of ephrinB1 is also required for this critical tyrosine-phosphorylation-dependent event. PMID- 14535846 TI - Identification of sequences required for the import of human protoporphyrinogen oxidase to mitochondria. AB - Protoporphyrinogen oxidase (PPOX; EC 1.3.3.4), the penultimate enzyme of haem biosynthesis, is a nucleus-encoded flavoprotein strongly associated with the outer surface of the inner mitochondrial membrane. It is attached to this membrane by an unknown mechanism that appears not to involve a membrane-spanning domain. The pathway for its import to mitochondria and insertion into the inner membrane has not been established. We have fused human PPOXs containing N terminal deletions, C-terminal deletions or missense mutations to yellow fluorescent protein (YFP) and have used these constructs to investigate the mitochondrial import of PPOX in human cells. We show that all the information required for efficient import is contained within the first 250 amino acid residues of human PPOX and that targeting to mitochondria is prevented by fusion of YFP to the N-terminus. Deletion of between 151 and 175 residues from the N terminus is required to abolish import, whereas shorter deletions impair its efficiency. Fully efficient targeting appears to require both a major targeting signal, the whole or part of which is contained between residues 151 and 175, and which may be involved in anchoring to the inner mitochondrial membrane, together with interaction between this region and a sequence(s) within the first 150 residues. These features suggest that the mechanism for import of human PPOX to mitochondria differs from those identified for the translocation of nucleus encoded, membrane-spanning, inner membrane proteins. In addition, a missense mutation outside this region (Val(335)-->Gly) prevented targeting to mitochondria and delayed the appearance of YFP fluorescence. This mutation appeared to prevent import by a direct effect on protein folding rather than by altering a sequence required for targeting. It may lead to sequestration of the PPOX-YFP construct in an unfolded conformation, followed by proteolytic degradation, possibly through enhanced binding to a cytosolic chaperone protein. PMID- 14535845 TI - Molecular basis of the high-affinity activation of type 1 ryanodine receptors by imperatoxin A. AB - Both imperatoxin A (IpTx(a)), a 33-residue peptide toxin from scorpion venom, and peptide A, derived from the II-III loop of dihydropyridine receptor (DHPR), interact specifically with the skeletal ryanodine receptor (RyR1), which is a Ca(2+)-release channel in the sarcoplasmic reticulum, but with considerably different affinities. IpTx(a) activates RyR1 with nanomolar affinity, whereas peptide A activates RyR1 at micromolar concentrations. To investigate the molecular basis for high-affinity activation of RyR1 by IpTx(a), we have determined the NMR solution structure of IpTx(a), and identified its functional surface by using alanine-scanning analogues. A detailed comparison of the functional surface profiles for two peptide activators revealed that IpTx(a) exhibits a large functional surface area (approx. 1900 A(2), where 1 A=0.1 nm), based on a short double-stranded antiparallel beta-sheet structure, while peptide A bears a much smaller functional surface area (approx. 800 A(2)), with the five consecutive basic residues (Arg(681), Lys(682), Arg(683), Arg(684) and Lys(685)) being clustered at the C-terminal end of the alpha-helix. The functional surface of IpTx(a) is composed of six essential residues (Leu(7), Lys(22), Arg(23), Arg(24), Arg(31) and Arg(33)) and several other important residues (His(6), Lys(8), Arg(9), Lys(11), Lys(19), Lys(20), Gly(25), Thr(26), Asn(27) and Lys(30)), indicating that amino acid residues involved in RyR1 activation make up over the half of the toxin molecule with the exception of cysteine residues. Taken together, these results suggest that the site where peptide A binds to RyR1 belongs to a subset of macrosites capable of being occupied by IpTx(a), resulting in differing the affinity and the mode of activation. PMID- 14535850 TI - Epidemiology and demographics of prostatitis. AB - Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is a multifactorial problem affecting men of all ages and demographics. Currently, there is a relative dearth of epidemiological information on CPPS. It is clear that patients with CPPS have a dismal quality of life and many have benefited only minimally from empiric, goal-directed therapy. Long-term follow-up of the CPPS cohort will answer important questions about the natural and treated history of this syndrome. Similarly, ongoing and future studies will provide community-based and prevalence estimates for CPPS, morbidity rates for men with CPPS, and the rates of symptom improvement and symptom deterioration for these men, as well as the probability of benefits and harm from different treatments. Although men with CP routinely receive anti-inflammatory and antimicrobial therapy, recent studies suggest that leucocyte and bacterial counts do not correlate with severity of symptoms. These findings suggest that factors other than leucocytes and bacteria contribute to the symptoms associated with CPPS. The probability of benefits and harm from different treatments for CPPS, and reliable and valid measures to define these outcomes are eagerly awaited. PMID- 14535847 TI - Fatty acids enhance membrane permeabilization by pro-apoptotic Bax. AB - Fatty acids are known promoters of apoptosis. In the present study, the direct role of fatty acids with regard to their ability to cause membrane permeabilization by Bax was explored. Addition of fatty acids to liposomes in the presence of cations greatly enhanced the permeabilizing activity of Bax, a pro apoptotic Bcl-2 protein. This provides a putative mechanism for the role of fatty acids in apoptosis. It is not a result of detergent-like properties of fatty acids, since a different micelle-forming amphiphile, dilysocardiolipin, was strongly inhibitory. We also demonstrate that there is a synergistic effect on Bax-induced permeabilization between Ca(2+) and Mg(2+), both on the binding of Bax to liposomes as well as on the induction of the leakage of liposomal contents. Micromolar concentrations of Ca(2+) added externally or submicromolar concentrations of free Ca(2+) present in the medium were sufficient to promote Bax-induced permeabilization synergistically with externally added Mg(2+). These results indicate that Bax can induce leakage from liposomes at ion concentrations resembling those found physiologically. The synergistic effects of Ca(2+) and Mg(2+) were observed with liposomes with different lipid compositions. Thus the action of Bax is strongly modulated by the presence of bivalent cations that can act synergistically, as well as by micelle-forming lipid components that can be either stimulatory or inhibitory. PMID- 14535848 TI - A membrane proximal region of the integrin alpha5 subunit is important for its interaction with nischarin. AB - In a previous study [Alahari, Lee and Juliano (2000) J. Cell Biol. 151, 1141 1154], we have identified a novel protein, nischarin, that specifically interacts with the cytoplasmic tail of the alpha5 integrin subunit. Overexpression of this protein profoundly affects cell migration. To examine the nischarin-alpha5 interaction in detail, and to find the minimal region required for the interaction, several mutants of nischarin and alpha5 were created. The results obtained for the yeast two-hybrid system indicate that a 99-aminoacid region of nischarin (from residues 464 to 562) is indispensable for the interaction. Also, we demonstrate that the membrane proximal region (from residues 1017 to 1030) of the alpha5 cytoplasmic tail is essential for the interaction. To characterize more directly the properties of the interaction between nischarin and alpha5, we performed surface-plasmon resonance studies in which peptides were immobilized on the surface of a sensor chip, and the recombinant nischarin protein fragments were injected. Consistent with the two-hybrid results, recombinant nischarin binds well to immobilized alpha5 peptides. In addition, mutational analysis revealed that residues Tyr(1018) and Lys(1022) are crucial for alpha5-nischarin interactions. These results provide evidence that nischarin is capable of directly and selectively binding to a portion of the alpha5 cytoplasmic domain. Further studies demonstrated that the minimal alpha5 binding region of nischarin does not affect cell migration. PMID- 14535851 TI - The 2001 Giessen Cohort Study on patients with prostatitis syndrome--an evaluation of inflammatory status and search for microorganisms 10 years after a first analysis. AB - During the last years tremendous changes have occurred in the epidemiologic knowledge and the diagnostic process of the prostatitis syndrome. A new worldwide accepted classification system has become the gold standard in contemporary literature. The aim of this study was to compare the inflammatory and infectious status of men with prostatitis syndrome with results from our study cohort from 1992. A total of 168 symptomatic men (mean age 43.2 years; range 18-79) attending the Giessen prostatitis outpatient department were included. All men underwent a standard four-glass-test including leucocyte analysis in all specimens. A routine search for Ureaplasma urealyticum and Chlamydia trachomatis was performed. Ejaculate analysis following World Health Organization (WHO) criteria has been performed including the evaluation of increased number of peroxidase-positive leucocytes (PPL). Men were classified according to the National Institutes of Health (NIH) prostatitis classification. The distribution of patients according to NIH criteria is as follows: NIH II (4.2%), NIH IIIA (31.5%), NIH IIIB (50.0%) and urethroprostatitis (14.3%). Chlamydial infection was present in one man (0.6%). Only two men with increased leucocytes in prostatic secretions demonstrated > or =106 million ml-1 PPL in semen. As compared with our cohort study 10 years ago, the proportion of the different subtypes of the prostatitis syndrome have remained stable. The aetiological spectrum of chronic bacterial prostatitis has not changed whereas, in contrast, the prevalence of C. trachomatis now is found to be strikingly reduced. Using the WHO cutpoints for leucocytospermia the inclusion of seminal leucocytes to the diagnostic process has not influenced the distribution between inflammatory (type NIH IIIA) and noninflammatory (type NIH IIIB) chronic pelvic pain syndrome. PMID- 14535852 TI - Chlamydial and ureaplasmal infections in patients with nonbacterial chronic prostatitis. AB - Our study was designed to establish the necessity of routine evaluation of patients with inflammatory (IIIA) and noninflammatory (IIIB) types of nonbacterial prostatitis (NBP) for chlamydial and ureaplasmal infections. From 1999 to 2001, 165 patients with a mean age of 35 years (range 20-54 years) were evaluated for the syndrome of chronic prostatitis. The evaluation included scoring with Prostate Symptom Score Index (PSSI) and NIH Chronic Prostatitis Symptom Index (CPSI), Meares-Stamey test and culturing of post-massage urine portion (fourth glass). In all cases, polymerase chain reaction (PCR)-testing of the semen was performed to establish the persistence of Chlamydia trachomatis (ChT) and Ureaplasma urealyticum (UU). Based on laboratory findings (four glass test and post-massage urine culture), in 69 (42%) of 165 cases, NBP was diagnosed, which includes 30 patients with type IIIA and 39 with type IIIB of NBP. According to semen PCR tests, in 11 (36.6%) of 30 cases with IIIA type of NBP, chlamydial (six cases), ureaplasmal (four cases) and a mixture of both (one case) infections were described. Among 39 patients with IIIB type of NBP test was positive in 14 cases (36%), where UU was presented in eight and ChT in six cases. In patients with previously diagnosed inflammatory as also noninflammatory NBP, according to four glass test, chlamydial and/or ureaplasmal infections can be presented. Although their role in pathogenesis of prostatitis remains speculative, however, testing for infections is highly recommended. PMID- 14535853 TI - Seminal fluid analysis in chronic prostatitis/chronic pelvic pain syndrome. AB - Prostatitis is a common cause of morbidity among adult men. There are more than 2,000,000 doctor visits per year in the United States, approximately half to urologists (Collins et al., 1998, J Urol 159:1224; Roberts et al., 1998, Urology 51:578; Krieger et al., 2003, Urology). The problem is that very few patients have obvious infections, or functional or structural abnormalities. The aim of this study is to examine our experience with seminal fluid analysis in this patient population, and to outline the potential utility of this examination in patient evaluation. PMID- 14535854 TI - The limit of leucocytospermia from the microbiological viewpoint. AB - The aim of the study was to find out the correlation between white blood cell (WBC) counts in semen and quantitative composition of seminal microflora, and to establish the minimum WBC count associated with significant bacteriospermia. The research included 159 men with different WBC counts in their semen, 84 of them with chronic prostatitis/chronic pelvic pain syndrome. Semen samples were cultivated quantitatively for detecting anaerobic, microaerophilic and aerobic bacteria. Bryan-Leishman stained slides were used for detecting WBC in semen. Seminal fluid was colonized by eight different microorganisms, and the total count of microorganisms in semen ranged from 102 to 107 CFU ml-1. A high frequency of anaerobic microorganisms was found. A positive correlation was observed between the WBC count and the number of different microorganisms, and also between the WBC count and the total count of microorganisms in semen sample. The receiver operating characteristic curve analysis demonstrated that the WHO defined WBC cut-off point (1 x 106 WBC ml-1) has very low sensitivity for discriminating between patients with and without significant bacteriospermia, as a more optimal sensitivity/specificity ratio appears at 0.2 x 106 WBC ml-1 of semen. The quantitative microbiological finding of semen in the patients of National Institute of Health (NIH) categories IIIa and IV was very similar, i.e. a high number of different microorganisms and a high total count of microorganisms. In the control group (without leucocytospermia and prostatitis symptoms) both parameters were significantly lower. PMID- 14535855 TI - Cellular and biochemical markers in semen indicating male accessory gland inflammation. AB - Leucocytospermia is considered to be a sign of male accessory gland inflammation. The leucocytes in semen are mainly polymorphonuclear neutrophilic granulocytes. Leucocytospermia is not associated with the presence of bacteria and antibiotic treatment does not significantly lower the extent of leucocytospermia. A higher frequency of elevated herpes simplex antibodies titres were found in men with leucocytospermia. The concentration of inflammatory cytokines, interleukin-6 and 8, is closely correlated with the number of leucocytes. Their determination does not provide additional information. Reactive oxygen species (ROS) are generated at least in part by seminal leucocytes in response to stimulating factors. Purified leucocytes produce high levels of ROS. The determination of ROS appears to represent a parameter of functional activity of leucocytes. The role of chlamydiae in male accessory gland infection is unclear. Their determination in semen by DNA amplification and by immunological tests does not provide reliable results. PMID- 14535856 TI - Urogenital infection and sperm motility. AB - Male accessory sex gland infections are considered as potential hazards to male fertility. Various pathophysiological concepts have evolved from experimental and clinical studies that begin to explain the effects of bacteria and immunological events on the function of spermatozoa and sperm motility in particular. Besides direct influences of pathogenic bacteria on spermatozoa whose impact on the motility of human spermatozoa is reviewed herein, recent studies have identified and evaluated infectious mediators that appear to be responsible for specific molecular processes in infections that particularly affect the motility of spermatozoa. This review will focus in detail on direct bacterial effects of sperm motility, the role of seminal leucocytes and the impact of pro-inflammatory cytokines on the motility of spermatozoa. PMID- 14535857 TI - Morphological sperm alternations in different types of prostatitis. AB - The possible negative effect of the chronic pelvic pain syndrome (CPPS) on semen parameters and especially sperm morphology is still a controversial matter. The aim of this study was therefore to investigate if different types of prostatitis can have a negative effect on sperm morphology and to compare our results with that of a literature survey. Semen analyses were performed on 34 males with confirmed CPPS (NIH III A), 18 males with CPPS (NIH III B) and 17 males as controls. When sperm morphology was evaluated according to WHO criteria, no differences were found between the mean percentages of morphologically normal spermatozoa for the three groups. An extended sperm morphology evaluation according to strict criteria showed that the NIH III A group had a tendency for a lower percentage of morphologically normal spermatozoa (5.3 +/- 3.1%) and acrosome index (8.7 +/- 4.8%) compared with the control group with values of 7.3 +/- 5.6% and 12.7 +/- 7.3%, respectively. There was a statistically significant higher (P = 0.0186) mean percentage (17.5 +/- 15.7%) of elongated spermatozoa in the NIH III A group compared with the control group (7.2 +/- 9.5%) while the NIH III B group had values between those of the control and NIH III A group. Our results indicate that CPPS NIH III A can have a significant negative effect on sperm morphology parameters, as evaluated by strict criteria, and to a lesser extent in cases of CPPS NIH III B compared with a control group. PMID- 14535858 TI - Immunological alterations in the ejaculate of chronic prostatitis patients: clues for autoimmunity. AB - The aim of this prospective study was to observe immunophenotypic patterns in the ejaculate of patients with noninflammatory chronic pelvic pain syndrome (Cat IIIB CPPS) and to test for a possible autoimmune aetiology. Thirty-five patients of a total of 88 patients with chronic prostatitis Cat IIIB were consecutively selected. Monthly ejaculate testing was carried out for IgG, IgA, IgM, IL-1alpha, sIL-2R and IL-6. The control group for ejaculate analysis was composed of 96 normal ejaculates (according to the WHO criteria). Immunohistochemical detection of CD3 cells (T lymphocytes) and CD20 cells (B lymphocytes) was performed in 71 biopsy cylinders of Cat IIIB CPPS patients and in 25 prostate biopsy cylinders of subjects without symptoms or obstruction. Intra-acinar T-lymphocytic infiltrates were dominated by T-cytotoxic cells (P = 0.05). Ejaculate IL-6 and ejaculate IgA increased significantly and dropped again, correlating with a release of clinical symptoms. Inflammatory ejaculate interleukin concentrations correlated with the immunohistochemical findings with presence of large numbers of T cells (all P values < or = 0.01). Immunomodulation was performed in a pilot series of three patients by five monthly cycles of IgG (Sandoglobulin), 1 g kg-1 body weight. Immunomodulation with IgG decreased pain moderately and did not change ejaculate interleukin and immunoglobulin concentrations. In summary, interleukin and immunoglobulin determinations in the ejaculate revealed an inflammatory process even in Cat IIIB CPPS. The findings of intra-acinar T-cell rich infiltrates and the associated inflammatory reaction may indicate a possible autoimmune component in the aetiology of CPPS. Exact origin and role of interleukin changes in the ejaculate of CPPS patients need to be further evaluated. Unfortunately, pilot series with immunomodulation with IgG do not seem to provide clear clinical benefit. PMID- 14535859 TI - Male accessory gland infection: standardization of inflammatory parameters including cytokines. AB - Assessment of infection of the male accessory glands is usually based on the search for white blood cells in different specimens to document an inflammatory reaction. This widely used practice allows to establish the diagnosis of inflammation in many cases. However, clinical symptoms do not always correlate with the presence or absence of white blood cells. This is particularly true for chronic prostatitis/chronic pelvic pain syndrome. In the last few years different research efforts have been made to look for markers of inflammation other than elements of the white blood cell line. Several studies suggest that humoral rather than cellular parameters are involved in male accessory gland infections. Substances such as reactive oxygen species, nerve growth factor and cytokines seem to be important not only in the pathogenesis of the inflammatory reaction but may also serve as diagnostic markers to indicate the presence of inflammation. PMID- 14535860 TI - Seminal oxidative stress in patients with chronic prostatitis. AB - Prostatitis has been associated with abnormal semen parameters and may be the cause of infertility in some patients. Sperm antibodies and impaired sperm motility have been observed in the semen of patients with prostatitis. More recently, seminal oxidative stress has been detected in patients with prostatitis or accessory gland infection. Oxidative stress is the result of elevated Reactive Oxygen Species (ROS) or depressed Total Anti-oxidant Capacity, or both. Granulocytes, in prostatic fluid may be responsible for the generation of ROS, which are known to impair sperm function. Abnormal levels of ROS, however, have also been detected in the absence of leucocytospermia. Several investigators have observed significant oxidative stress in the semen or prostatic fluid from patients with bacterial and abacterial forms of prostatitis, comparable with levels reported in men with recognized infertility. Whether prostatitis is a risk factor for infertility or not, remains controversial. Therefore, the role of seminal oxidative stress as either the result of prostatic inflammation or the cause of infertility is likewise debatable, but plausible. We summarize the results of our studies, as well as those of others who are dedicated to the research of prostatitis, and its potential effects on semen quality. PMID- 14535861 TI - Urogenital inflammation: changes of leucocytes and ROS. AB - The presence of excess leucocytes in the semen has been associated with male infertility. According to the WHO, concentrations of more than 106 leucocytes ml 1 are considered as leucocytospermia, indicating genital tract infections. Up to now, no consensus has been achieved on how leucocytes should be quantified in semen. Using the peroxidase staining and monoclonal antibodies to CD15, CD45 and CD68, we found significant differences between the detection methods. Only 47.4% of the semen samples that were assessed as leucocytospermic by CD45 were identified as such by peroxidase staining. The concentration of peroxidase positive cells was significantly correlated with polymorphonuclear granulocyte (PMN) elastase (P < 0.0001). However, a negative correlation of peroxidase positive cells with the sperm concentration was only found in oligozoospermic patients (P < 0.0001). Moreover, the slightly positive correlation with normal sperm morphology seems to be applicable only in cases of oligozoospermia. Significant negative correlation of the number of peroxidase-positive cells were found for both maximal inducible acrosome reaction (P = 0.0219) and the inducibility of acrosome reaction (P = 0.0370), indicating a rather deleterious effect of leucocytes on this important sperm function. Concerning the result in the in vitro fertilization programme, none of the examined parameters (PMN elastase, concentration of round cells and peroxidase-positive cells) showed a correlation with either fertilization or pregnancy. This result seems to be reasonable as severely damaged spermatozoa and leucocytes are eliminated from the ejaculate by different sperm separation methods. Interestingly, a significant negative correlation of the TUNEL assay as a measure of sperm DNA fragmentation was found only with pregnancy (P = 0.006) but not with fertilization. As DNA fragmentation can also be caused by ROS that are generated by leucocytes, this causality should not be neglected. PMID- 14535862 TI - Acrosome reaction in Chlamydia-positive and negative patients. AB - Chlamydia trachomatis infections might have a detrimental effect on various sperm functions. Data concerning the effect of C. trachomatis on the capacitation activity of sperms are lacking. The study was undertaken to evaluate whether chlamydial infection influences acromsome reaction (AR). Three groups of men were investigated for ARs -Chlamydia negative (n = 46) and positive (n = 30) patients, and healthy men (n = 53) undergoing vasectomy. The fluorescence technique for the evaluation of AR was applied. The normal range for the induction of AR was assumed DeltaAR > 12.5% for this technique. Seminal plasma was examined for IgA antibodies against C. trachomatis. There was a significant difference in AR between healthy volunteers, Chlamydia-negative and Chlamydia-positive patients. DeltaARs were 15.8 +/- 1.6% in healthy volunteers versus 12.15 +/- 2.4% in Chlamydia-negative and 9.08 +/- 1.8% in Chlamydia-postive patients, respectively (P < 0.05). Significant elevated titres of C. trachomatis-specific IgA in seminal plasma showed a negative correlation with the AR of spermatozoa. AR seems to be a valuable marker, especially in couples with idiopathic infertility. PMID- 14535863 TI - Aetiology of haemospermia. AB - There are several unknown factors which cause haemospermia. An earlier developed diagnostic scheme has been expended by novel imaging techniques and biopsy methods. A detailed case history, physical examination and microscopic analysis of the ejaculate is required. In haemo-pyospermia a complete microbiological analysis must be escalated. Noninvasive imaging techniques (ultrasound, computer tomography and magnetic resonance imaging) help in detecting calculous and malignant diseases. So far, as a precise diagnosis has not been available, urethroscopy has been performed. Malignancies (prostate, seminal vesicles) must be histologically verified by biopsies. In contempt of our efforts the practice shows a part of haemospermia remaining essential. Analysing two time periods we found prostatic calculi, chronic prostatitis and carcinoma of the prostate unequivocally as most frequent causes. Considering the rare genital malignancies we find more than 10% frequency. Notably, in our study only 2.4% of the malignancies occurred in patients under 40 years of age. Hence a detailed diagnosis is advocated in haemospermia patients over 40 years. Finally, we may state that in contempt of the applied modern imaging techniques 15% of patients with haemospermia had unknown aetiology. PMID- 14535864 TI - Inflammatory-associated obstructions of the male reproductive tract. AB - A history of urogenital inflammation occurs in 5-12% of men attending infertility clinics. Usually, infection has a detrimental effect on sperm quality by reducing concentration and motility, and possibly affecting the number of morphological normal spermatozoa. In addition, infection may be the source of auto-antibodies against spermatozoa, found in about 8% of the infertile male population. In contrast to the situation in women, there is no clear evidence that male accessory gland infections can result in epididymal blockage or vassal obstruction, with the exception of genital tuberculosis. Although Chlamydia trachomatis is a well-documented source of chronic prostatitis, the infection does not seem to cause obstruction of the reproductive tract, as it does in women. If male urogenital infection causes obstruction it is most likely located at the level of the ejaculatory ducts. Chronic prostatitis has been proved to cause scarring of the prostatic and ejaculatory ducts, resulting in low seminal volume with low fructose and alpha-glucosidase. Many of these men present with severe oligozoospermia or azoospermia, normal size testis and normal gonadotrophins. We performed an excisional testicular biopsy in all men presenting with <1 million spermatozoa per millilitre and found that 39 of 78 (50%) had a normal spermatogenesis. A history of male accessory genital infection was found in 12% of the men and 10% had abnormalities found on transrectal ultrasound of the prostate (like oedema, dilatation of the seminal vesicles and ejaculatory ducts) intraprostatic calcifications and dilatation of the periprostatic venous plexus. Ejaculatory duct obstruction is a common cause of male infertility and infections are present in at least 22-50% of these men. Transurethral resection of the ejaculatory ducts may result in a significant improvement of the sperm quality and in spontaneous pregnancies in up to 25% of the couples. In case of failure sperm aspiration from the epididymis and intracytoplasmic sperm injection is the treatment of choice. PMID- 14535865 TI - Chronic prostatitis and male accessory gland infection--is there an impact on male infertility (diagnosis and therapy)? AB - The aim of this article was to discuss by means of a review of the literature and own study material the multifactorial aetiology of male infertility, extrapolate this hypothesis to male accessory gland infection (MAGI) and relate it to chronic prostatitis and its treatment. Infertility is a multifactorial disease and diagnosis and therapy must be oriented as such. Although the relationship between prostatitis and infertility remains unclear, bacteria, viruses, leucocytes, reactive oxygen species, cytokines, obstruction and immunological abnormalities must be seen as cofactors in the development of infertility in patients with MAGI and prostatitis. Infection, trauma, allergy, neurogenic damage, chemical or mechanical factors can lead to a long-lasting inflammation of the prostate or pelvic organs even after eradication of the aetiological agent, and is potentially related to infertility through cytokines. In relation to treatment of infertility, antibiotics play a role in bacterial prostatitis whereas in abacterial prostatitis other treatments like antioxidants, sacral nerve stimulation and anti-inflammatory treatment are worth to be studied in the future. PMID- 14535866 TI - Antibiotic therapy--rationale and evidence for optimal drug concentrations in prostatic and seminal fluid and in prostatic tissue. AB - The theoretical background of drug penetration into the prostate is outlined, emphasizing the phenomenon of ion-trapping and the role of nonionic diffusion of weak acids, bases and amphoteric drugs across biological membranes with a pH gradient. Determination of drug concentrations in human prostatic secretion are problematic because of possible urinary contamination. Studies have been carried out mainly in healthy volunteers. The results have to be interpreted with caution, if not care was taken to rule out or at least identify urinary contamination. Analysing the concentrations of various fluoroquinolones in prostatic and seminal fluid as well as in prostatic tissue, it becomes obvious that the fluoroquinolones differ not only in plasma concentrations but also in their penetration ability to these sites. In spite of intensive investigations, our knowledge is still limited concerning the mechanisms that govern the transport of antibiotic drugs into and their activity in the various prostatic compartments and how the findings can be applied clinically. Nevertheless, overall the concentrations at the site of infection of most of the fluoroquinolones should be sufficient for the treatment of chronic bacterial prostatitis and vesiculitis caused by susceptible pathogens. PMID- 14535868 TI - Meta-analysis: proton pump inhibitors vs. H2-receptor antagonists--their efficacy with antibiotics in Helicobacter pylori eradication. AB - BACKGROUND: It is unknown whether proton pump inhibitors are superior to H2 receptor antagonists in Helicobacter pylori eradication regimens. AIM: To perform a meta-analysis comparing the efficacy of both antisecretors when co-prescribed with antibiotics. METHODS: Randomized clinical trials comparing proton pump inhibitors vs. H2-receptor antagonists with the same antibiotics were selected. Data sources included PubMed, the Cochrane Controlled Trials Register and abstracts from congresses up to January 2002. A meta-analysis was performed by combining the odds ratios. RESULTS: Twenty studies fulfilled the inclusion criteria. In the intention-to-treat analysis, the mean eradication rates with proton pump inhibitors and H2-receptor antagonists plus antibiotics were 74% [95% confidence interval (CI), 71-76%] and 69% (95% CI, 66-71%), respectively. The odds ratio for this comparison was 1.31 (95% CI, 1.09-1.58). The number needed to treat with proton pump inhibitors to achieve eradication success, compared with H2-receptor antagonists, was 25. When studies prescribing very high doses of H2 receptor antagonists (two of the outliers) were excluded, the odds ratio (for proton pump inhibitors vs. H2-receptor antagonists) increased to 1.37, the number needed to treat decreased to 20 and the heterogeneity between the studies decreased. CONCLUSIONS: Overall, proton pump inhibitors are more effective than H2-receptor antagonists when prescribed at usual doses with antibiotics to eradicate H. pylori infection. PMID- 14535869 TI - Quality of life in relation to symptoms in patients with gastro-oesophageal reflux disease-- an analysis based on the ProGERD initiative. AB - AIMS: To determine the impact of gastro-oesophageal reflux disease (GERD) on the quality of life, to assess changes in the quality of life during treatment with esomeprazole and to define factors that can predict these changes. METHODS: Patients with GERD (n=6215) were included in a prospective cohort study (ProGERD). All patients underwent endoscopy and received esomeprazole. At baseline and after 2 weeks of treatment, symptoms and quality of life were assessed. Factors that influenced changes in the quality of life were determined by multiple regression analyses. RESULTS: At baseline, the quality of life in GERD patients was lower than that in the general population, and was similar to that in patients after acute coronary events. No differences in symptoms or quality of life were observed between the subgroups of patients with non-erosive GERD, erosive GERD and Barrett's oesophagus. After treatment with esomeprazole, the symptoms and quality of life were improved in all subscales within 2 weeks (P<0.001). The mean score of the disease-specific quality of life instrument (Quality of Life in Reflux and Dyspepsia Patients) increased from 4.6 to 6.2 points, representing a highly relevant clinical improvement. The generic quality of life (SF-36) reached levels similar to those in the general population, but, again, no difference was found between the three different subgroups of GERD patients. The main factors associated with an improvement in the quality of life after treatment were symptom relief, severe erosive reflux disease, absence of extra-oesophageal disorders, avoidance of non-steroidal anti-inflammatory drug intake and positive Helicobacter pylori status. CONCLUSIONS: GERD causes a significant impairment in the quality of life that can be attenuated or normalized within a time period as short as 2 weeks by treatment with esomeprazole. These findings were similar across the whole GERD patient spectrum. PMID- 14535871 TI - 13C-methacetin breath test for monitoring hepatic function in cirrhotic patients before and after liver transplantation. AB - BACKGROUND: In patients with chronic liver disease, the measurement of liver function is critical for monitoring disease progression, predicting the prognosis and choosing therapeutic strategies. The 13C-methacetin breath test is a simple, non-invasive diagnostic tool based on an inexpensive, non-toxic substance, which allows the accurate measurement of liver functional reserve. AIM: To investigate the 13C-methacetin breath test as a tool to monitor hepatic function in liver transplant candidates and recipients. METHODS: Twenty-eight cirrhotic patients listed for orthotopic liver transplantation and 10 healthy controls were studied. The 13C-methacetin breath test (75 mg per os) was performed at baseline and at 12 week intervals. Intra-operative measurements were obtained during the liver transplantation procedure in nine patients. Results were expressed as the 13C methacetin cumulative oxidation percentage 45 min after substrate ingestion. RESULTS: The mean 13C-methacetin cumulative oxidation at 45 min was 16.4 +/- 3.5% in healthy controls and 5.4 +/- 4.2% in cirrhotic patients at the time of listing. In 11 patients who underwent successful liver transplantation, mean oxidation increased from 3.3 +/- 1.6% before transplantation to 17.0 +/- 5.2% at 6 months of follow-up. Variations in methacetine oxidation were closely related to the recovery of liver function. The mean intra-operative 13C-methacetin cumulative oxidation increased from 0.1% during the anhepatic phase to 3.7 +/- 2.0% 2 h after reperfusion. CONCLUSIONS: The 13C-methacetin breath test is a simple and potentially useful tool for monitoring hepatic function in cirrhotic patients listed for liver transplantation, and during the intra-operative and post-operative phases. PMID- 14535870 TI - Effect of propranolol and depot lanreotide SR on postprandial and circadian portal haemodynamics in cirrhosis. AB - BACKGROUND: Long-acting somatostatin analogues have been suggested as an alternative to propranolol for the prevention of variceal rebleeding. AIM: To compare the effectiveness of lanreotide SR, a new depot formulation injected once weekly, and propranolol in reducing circadian portal blood flow (PVF) and meal stimulated hepatic venous pressure gradient (HVPG) in patients with liver cirrhosis. METHODS: Patients were randomized to receive either lanreotide SR intramuscularly (30 mg once weekly, n=12) or propranolol (n=12) orally. Hemodynamic measurements were performed on day 0 and on day 21 after a 3-week period of drug administration, while patients received three standard oral liquid test meals. On each study day 27 PVF measurements were performed over 24 h and eight measurements of HVPG during the first postprandial period. RESULTS: Propranolol was more effective than lanreotide SR in reducing baseline HVPG ( 21.9 vs. -13.6%, P=0.04) and meal-stimulated HVPG (-16.6 vs. -3.8%, P=0.04). Propranolol reduced circadian PVF significantly by 9.3% (P=0.03) but not lanreotide SR. CONCLUSIONS: Long-term treatment with propranolol reduced baseline and postprandial HVPG and circadian PVF, while lanreotide SR did not. The results of our study do not encourage clinical testing of lanreotide SR 30 mg for the prevention of variceal haemorrhage. PMID- 14535872 TI - Second-line treatment for failure to eradicate Helicobacter pylori: a randomized trial comparing four treatment strategies. AB - AIM: To compare the efficacy of different regimens in patients in whom previous Helicobacter pylori eradication therapy has failed. METHODS: In this study named StratHegy patients (n=287) were randomized to receive one of three empirical triple therapy regimens or a strategy based on antibiotic susceptibility. The empirical regimens were omeprazole, 20 mg b.d., plus amoxicillin, 1000 mg b.d., and clarithromycin, 500 mg b.d., for 7 days (OAC7), clarithromycin, 500 mg b.d., for 14 days (OAC14) or metronidazole, 500 mg b.d., for 14 days (OAM14). In the susceptibility-based strategy, patients with clarithromycin-susceptible strains received OAC14, whilst the others received OAM14. The 13C-urea breath test was performed before randomization and 4-5 weeks after eradication therapy. RESULTS: In the intention-to-treat analysis, the eradication rates for empirical therapies were as follows: OAC7, 47.4% (27/57); OAC14, 34.5% (20/58); OAM14, 63.2% (36/57); it was 74.3% (84/113) for the susceptibility-based treatment (P<0.01 when compared with OAC7 and OAC14). In patients receiving clarithromycin, the eradication rates were 80% for clarithromycin-susceptible strains and 16% for clarithromycin-resistant strains; in patients receiving OAM14, the eradication rates were 81% for metronidazole-susceptible strains and 59% for metronidazole resistant strains. CONCLUSIONS: Eradication rates of approximately 75% can be achieved with second-line triple therapy based on antibiotic susceptibility testing. If susceptibility testing is not available, OAM14 is an appropriate alternative. PMID- 14535873 TI - Esomeprazole-based one-week triple therapy with clarithromycin and metronidazole is effective in eradicating Helicobacter pylori in the absence of antimicrobial resistance. AB - AIM: This study aimed to investigate the effectiveness of a one-week triple therapy with esomeprazole, clarithromycin and metronidazole for eradication of Helicobacter pylori infection in the absence of antimicrobial resistance. METHODS: Patients testing positive for H. pylori susceptible to metronidazole and clarithromycin (E-test) were randomized to receive a one-week regimen with either esomeprazole 2 x 20 mg or omeprazole 2 x 20 mg in combination with clarithromycin 2 x 250 mg and metronidazole 2 x 400 mg. Follow-up endoscopy with histology and culture and/or rapid urease test was performed 4-8 weeks after the end of treatment. RESULTS: Eighty patients were randomized. Helicobacter pylori infection was cured in 38/39 patients of the esomeprazole group and 31/33 patients of the omeprazole group (per protocol 97.4% (95% confidence interval [CI], 86.2-99.9), 93.7% (95% CI, 79.2-99.2), P=0.59); intention-to-treat 90.4% (95% CI: 77.4-97.3), 81.6% (95% CI: 65.7-92.3), respectively. No major side effects occurred. Minor side effects occurred in eight (20%) and six (23%) patients during esomeprazole and omeprazole therapy, respectively. Post-treatment susceptibility testing revealed resistance to both metronidazole and clarithromycin in two of the three patients who failed. CONCLUSION: We conclude that esomeprazole, clarithromycin and metronidazole as one-week triple therapy is effective for eradication of H. pylori in the absence of antimicrobial resistance. PMID- 14535874 TI - Favourable effect of regular intake of fermented milk containing Lactobacillus johnsonii on Helicobacter pylori associated gastritis. AB - BACKGROUND: Lactobacillus johnsonii (Lj1) had an in vitro and in vivo inhibitory effect on Helicobacter pylori. Fermented milk containing Lj1 (LC1), coadministered with antibiotics had a favourable effect on H. pylori gastritis. AIM: Evaluate the effect of LC1 intake without antibiotics on H. pylori gastritis. METHODS: Fifty H. pylori positive healthy volunteers were randomised in a double-blind study to LC1 or placebo. Gastric biopsies from the antrum and corpus were obtained before, and after 3 and 16 weeks of treatment, for histology and quantitative cultures. RESULTS: Severity and activity of antral gastritis was reduced after 16-week LC1 intake (pretreatment and 16-week inflammatory cell score: 6.0 +/- 0.8 vs. 5.3 +/- 0.1; P=0.04). H. pylori density decreased in the antrum after LC1 intake (3-week: 4.4 +/- 0.6; 16-week: 4.3 +/- 0.5 log10 colony forming units (cfu) vs. pretreatment 4.5 +/- 0.4 log10 cfu; P=0.04, respectively). Mucus thickness increased after 16 weeks of LC1 consumption (change of mucus thickness with LC1 and placebo in the antrum: 0.6 +/- 1.3 vs. 0.2 +/- 1.0, P=0.01; in the corpus: 0.3 +/- 1.1 vs. -0.6 +/- 1.5, P=0.03). CONCLUSION: LC1 intake had a favourable, albeit weak, effect on H. pylori associated gastritis, particularly in the antrum. Regular ingestion of fermented milk containing L. johnsonii may reduce the risk of developing disorders associated with high degrees of gastric inflammation and mucus depletion. PMID- 14535875 TI - Randomized study of different 'second-line' therapies for Helicobacter pylori infection after failure of the standard 'Maastricht triple therapy'. AB - BACKGROUND: Triple therapy with proton pump inhibitor, clarithromycin and amoxicillin and, in the event of eradication failure, quadruple therapy with proton pump inhibitor, bismuth, tetracycline and metronidazole have been proposed in Maastricht as the optimal sequential treatment of Helicobacter pylori infection. AIM: To compare two second-line regimens with quadruple therapy. METHODS: One hundred and eighty patients with a previous failed course of standard therapy were randomly given one of the following 7-day treatments: ranitidine bismuth citrate 400 mg b.d. plus amoxicillin 1 g b.d. and tinidazole 500 mg b.d. (RBCAT), pantoprazole 40 mg b.d. plus amoxicillin 1 g b.d. and levofloxacin 500 mg/day (PAL) and pantoprazole 40 mg b.d., bismuth citrate 240 mg b.d., tetracycline 500 mg q.d.s. and metronidazole 500 mg b.d. (PBTM). The eradication rate was assessed by 13C-urea breath test. Side-effects and compliance were evaluated by a standardized questionnaire and by counting returned medication. RESULTS: The RBCAT, PAL and PBTM groups achieved mean intention-to-treat eradication rates of 85%, 63% and 83%, respectively (P<0.05 for PAL vs. either RBCAT or PBTM). Compliance was optimal in all patients, although side-effects were more commonly observed in the PBTM group than in the other two patient groups (P<0.0001). CONCLUSIONS: Both RBCAT and PBTM can be used as second-line therapies. Conversely, PAL did not achieve satisfactory eradication rates. PMID- 14535876 TI - Role of antibiotic sensitivity testing before first-line Helicobacter pylori eradication treatments. AB - BACKGROUND: The resistance of Helicobacter pylori to antibiotics has been advocated as a major cause of treatment failure, and antimicrobial sensitivity testing has been proposed to improve efficacy; however, its role before first line therapy has not been investigated in detail. AIM: To assess whether antimicrobial sensitivity testing improves the eradication rate of first-line anti-Helicobacter treatments and to compare the effectiveness of ranitidine bismuth citrate and omeprazole in the presence of H. pylori resistance to antibiotics. METHODS: Two hundred and forty-two patients were assigned to either empirical or antimicrobial sensitivity testing-based treatment; within each group, subjects were further randomized to receive ranitidine bismuth citrate, 400 mg b.d., tinidazole, 500 mg b.d., and clarithromycin, 500 mg b.d., or omeprazole, 20 mg b.d., clarithromycin, 500 mg b.d., and amoxicillin, 1 g b.d., for 1 week, with substitution of the resistant antibiotic in the antimicrobial sensitivity testing-based treatment group. RESULTS: Eradication rates were 67% [confidence interval (CI), 55-79%] in the empirical treatment group and 76% (CI, 65-87%) in the antimicrobial sensitivity testing-based group (P=N.S.). The overall success rate was 60% (CI, 51-69%) with omeprazole and 82% (CI, 73-91%) with ranitidine bismuth citrate (P<0.03); the latter overcame antibiotic resistance in 12 of 15 strains vs. zero of eight strains by omeprazole. CONCLUSIONS: Antimicrobial sensitivity testing before first-line treatment does not improve the eradication rate, which is greater when ranitidine bismuth citrate is included in the treatment. PMID- 14535877 TI - Lansoprazole reduces ulcer relapse after eradication of Helicobacter pylori in nonsteroidal anti-inflammatory drug users--a randomized trial. AB - AIM: To study whether prophylaxis with lansoprazole could prevent relapse of ulcers after eradication of Helicobacter pylori in patients with NSAID-related peptic ulcers. METHODS: Patients who presented with peptic ulcers and were found to be infected with H. pylori while receiving NSAIDs were recruited into the study. They received, twice daily, lansoprazole 30 mg, amoxicillin 1 g and clarithromycin 500 mg for 1 week, followed by lansoprazole 30 mg daily for 4 weeks. Patients with healed ulcers and H. pylori eradicated were given naproxen 750 mg daily, and randomly assigned to receive lansoprazole 30 mg daily or no treatment for 8 weeks. The primary endpoint was the cumulative recurrence of symptomatic and complicated ulcers. RESULTS: At the end of the 8-week treatment period, significantly fewer patients (1/22, 4.5%, 95% confidence interval [CI] 0 23) in the lansoprazole group compared with the group that received H. pylori eradication alone (9/21, 42.9%, 95% CI 22-66) developed recurrence of symptomatic and complicated ulcers (log rank test P=0.0025). CONCLUSIONS: Lansoprazole significantly reduced the cumulative relapse of symptomatic and complicated ulcers in patients requiring NSAIDs after eradication of H. pylori. PMID- 14535878 TI - Expression of carbonic anhydrase IX in human pancreatic cancer. AB - BACKGROUND: Carbonic anhydrase IX has been linked to cancer development and progression. AIM: To analyse carbonic anhydrase IX expression and anhydrase inhibition in pancreatic cancer and to correlate these findings with p53 expression and microvessel density. MATERIALS AND METHODS: Seventy-seven pancreatic cancers were examined (43 males, 34 females; mean age, 64 years). The anti-carbonic anhydrase IX M75 antibody was used for immunohistochemistry and Western blot analysis. Microvessels were visualized using the anti-CD34 antibody, and p53 expression in cancer cells was assessed with a specific anti-p53 antibody. Quantitative polymerase chain reaction was performed in order to assess carbonic anhydrase IX mRNA levels in the pancreas. Furthermore, pancreatic cancer cell lines were treated with acetazolamide, a carbonic anhydrase inhibitor. RESULTS: In the normal pancreas, carbonic anhydrase IX immunoreactivity was observed at the basolateral membrane of ductal cells in 24 cases (31%). Carbonic anhydrase IX expression was found at the membrane and in the cytoplasm of pancreatic cancer cells in 16 pancreatic cancers (21%). Carbonic anhydrase IX expression was independent of the localization, stage, size, metastases and differentiation of the tumour. p53 expression was significantly more frequent in poorly differentiated cancers (P=0.0323); however, p53 expression and microvessel density were independent of carbonic anhydrase IX expression. Overall, carbonic anhydrase IX expression was not altered in pancreatic cancers vs. adjacent normal pancreatic tissue as assessed by Western blot and quantitative polymerase chain reaction analysis. However, incubation of pancreatic cancer cell lines with acetazolamide led to a significant inhibition of cell proliferation in AsPC-1 and PANC-1 pancreatic cancer cells. CONCLUSION: Carbonic anhydrase IX expression is observed in both ductal epithelial and cancer cells of the pancreas. Although the expression of carbonic anhydrase IX in pancreatic cancer is not associated with angiogenesis or advanced disease, it may well be a target for carbo-anhydrase inhibitors in a subset of pancreatic cancers. PMID- 14535879 TI - Gastric acid-suppressive therapy and community-acquired respiratory infections. AB - BACKGROUND: Bacteria and viruses have been detected in the stomach of patients during acid-suppressive therapy. AIM: To investigate whether subjects using acid suppressive drugs more often develop community-acquired respiratory infections when compared to those who do not use acid-suppressive drugs. METHODS: 700 study subjects were recruited during a single week in December 2002. Information on the prevalence of clinical manifestations of infections and complications in the preceding month was assessed by questionnaire. Furthermore, subjects were asked to report antibiotic therapy and physician visits related to possible infection. RESULTS: Questionnaires were returned by 405 subjects (58%). Consumption of acid suppressive drugs was reported by 91 individuals, of whom 79 used proton-pump inhibitors (20%) and 12 H2-receptor antagonists (3%). Overall, 101 (25%) responders reported clinical manifestations of respiratory infection in the preceding month. Subjects using acid-suppressive drugs were 2.34 times [95% confidence interval (CI) 1.4-4.1] more likely to have clinical manifestations of infection than individuals not using acid-suppressive drugs. Subjects using acid suppressive drugs visited a physician 3.72 times more often (95% CI 2.1-6.8) for an infection and received antibiotic therapy 4.19 times more often (95% CI 2.2 8.1) in comparison to individuals not using acid-suppressive drugs. CONCLUSIONS: Subjects using acid-suppressive drugs more often reported community-acquired respiratory infections in comparison to those who did not use acid-suppressive drugs. PMID- 14535880 TI - Mass spectrometric approach for identifying putative plasma membrane proteins of Arabidopsis leaves associated with cold acclimation. AB - Although enhancement of freezing tolerance in plants during cold acclimation is closely associated with an increase in the cryostability of plasma membrane, the molecular mechanism for the increased cryostability of plasma membrane is still to be elucidated. In Arabidopsis, enhanced freezing tolerance was detectable after cold acclimation at 2 degrees C for as short as 1 day, and maximum freezing tolerance was attained after 1 week. To identify the plasma membrane proteins that change in quantity in response to cold acclimation, a highly purified plasma membrane fraction was isolated from leaves before and during cold acclimation, and the proteins in the fraction were separated with gel electrophoresis. We found that there were substantial changes in the protein profiles after as short as 1 day of cold acclimation. Subsequently, using matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS), we identified 38 proteins that changed in quantity during cold acclimation. The proteins that changed in quantity during the first day of cold acclimation include those that are associated with membrane repair by membrane fusion, protection of the membrane against osmotic stress, enhancement of CO2 fixation, and proteolysis. PMID- 14535881 TI - Formation of plant cuticle: evidence for the occurrence of the peroxygenase pathway. AB - Cuticle plays a major role as a protective barrier in plants. Despite its physiological importance, the mode of formation of this complex structure remains poorly understood. In particular, none of the putative enzymes involved in the biosynthesis of the cutin, the matrix of cuticle, have been cloned. We have shown previously that peroxygenase is able to catalyze in vitro the epoxidation step required for the biosynthesis of C18 cutin monomers. In the present work, we have confirmed in planta that this oxidase is indeed a key enzyme involved in the formation of cutin. Thus, in maize leaves, the specific inactivation of peroxygenase by organophosphorothioates resulted in a dramatic decrease of cuticular epoxide content, as visualized by a specific histochemical technique that was accompanied by a reduced thickness of the cuticle. A strict correlation could also be established between the extent of inhibition of the peroxygenase and the modification of the cuticle triggered by a family of structurally related inhibitors. Importantly, these effects were restricted to plants that contain a cutin originating from C18 monomers. The altered cuticle of maize, treated with the peroxygenase inhibitor, was characterized by an increased permeability to pesticides. In addition, such plants became largely susceptible to infection by fungi, implying that the cuticle represents a crucial target for the modulation of the response in plant-pathogen interactions. PMID- 14535882 TI - Enhanced formation of flowers in salt-stressed Arabidopsis after genetic engineering of the synthesis of glycine betaine. AB - Previously, we showed that transformation with the codA gene for choline oxidase allows plants to synthesize glycine betaine (GB) and enhances their ability to tolerate various kinds of stress during germination and vegetative growth. In this study, we examined the tolerance of transformed plants to salt stress at the reproductive stage, which is the stage at which plants are most sensitive to environmental stress. Salt-shock treatment of wild-type plants for 3 days resulted in the abortion of flower buds and decreased the number of seeds per silique. These deleterious effects were clearly visible 6 days after the termination of salt-shock treatment. Microscopic examination of floral structures revealed that salt stress inhibited the development of anthers, pistils, and petals. In particular, the production of pollen grains and ovules was dramatically inhibited. These effects of salt stress were significantly reduced by transformation with the codA gene, and our observations suggested that the enhanced tolerance of the transgenic plants was a result of the accumulation of GB in the reproductive organs. Indeed, levels of GB in flowers, siliques, and inflorescence apices were about five times higher than in leaves. PMID- 14535883 TI - Transcriptome analysis of Arabidopsis colonized by a plant-growth promoting rhizobacterium reveals a general effect on disease resistance. AB - RNA transcript levels of Arabidopsis plants, infected by the rhizobacterium Pseudomonas thivervalensis (strain MLG45), and axenic control plants were compared using cDNA microarrays representing approximately 14 300 genes. The analysis revealed an increase of defence-related transcripts in the shoots of bacterized plants relative to control (axenic) plants. These modifications of transcript levels were confirmed by physiological experiments. Plants infected with P. thivervalensis were more resistant to subsequent infections by the virulent pathogen P. syringae pv. tomato (strain DC3000) than control plants. In addition, photosynthesis rates were repressed consistently with the reduced growth of plants colonized by P. thivervalensis. These results highlight the value of molecular phenotyping to predict physiological changes. PMID- 14535884 TI - Roles of OsCKI1, a rice casein kinase I, in root development and plant hormone sensitivity. AB - Casein kinases are critical in cell division and differentiation across species. A rice cDNA fragment encoding a putative casein kinase I (CKI) was identified via cDNA macroarray under brassinosteroid (BR) treatment, and a 1939-bp full-length cDNA, OsCKI1, was isolated and found to encode a putative 463-aa protein. RT-PCR and Northern blot analysis indicated that OsCKI1 was constitutively expressed in various rice tissues and upregulated by treatments with BR and abscisic acid (ABA). Enzymatic assay of recombinant OsCKI1 proteins expressed in Escherichia coli showed that the protein was capable of phosphorylating casein. The physiological roles of OsCKI1 were studied through antisense transgenic approaches, and homozygous transgenic plants showed abnormal root development, including fewer lateral and adventitious roots, and shortened primary roots as a result of reduced cell elongation. Treatment of wild-type plants with CKI-7, a specific inhibitor of CKI, also confirmed these functions of OsCKI1. Interestingly, in transgenic and CKI-7-treated plants, exogenously supplied IAA could restore normal root development, and measurement of free IAA content in CKI deficient primary and adventitious roots revealed altered auxin content, indicating that OsCKI1 is involved in auxin metabolism or that it may affect auxin levels. Transgenic plants were less sensitive than control plants to ABA or BR treatment during germination, suggesting that OsCKI1 may be involved in various hormone-signaling pathways. OsCKI1-GFP fusion studies revealed the localization of OsCKI1 to the nucleus, suggesting a possible involvement in regulation of gene expression. In OsCKI1-deficient plants, differential gene expression was investigated using cDNA chip technology, and results indicated that genes related to signal transduction and hormone metabolism were indeed with altered expression. PMID- 14535885 TI - Genomic and physiological studies of early cryptochrome 1 action demonstrate roles for auxin and gibberellin in the control of hypocotyl growth by blue light. AB - Blue light inhibits elongation of etiolated Arabidopsis thaliana hypocotyls during the first 30 min of irradiation by a mechanism that depends on the phototropin 1 (phot1) photoreceptor. The cryptochrome 1 (cry1) photoreceptor begins to exert control after 30 min. To identify genes responsible for the cry1 phase of growth inhibition, mRNA expression profiles of cry1 and wild-type seedlings were compared using DNA microarrays. Of the roughly 420 genes found to be differentially expressed at the point of cry1 response incipience, approximately half were expressed higher and half lower in cry1 relative to the wild type. Many of the cry1-dependent genes encoded kinases, transcription factors, cell cycle regulators, cell wall metabolism enzymes, gibberellic acid (GA) biosynthesis enzymes, and auxin response factors. High-resolution growth studies supported the hypothesis that genes in the last two categories were indeed relevant to cry1-mediated growth control. Inhibiting GA4 biosynthesis with a 3beta-hydroxylase inhibitor (Ca-prohexadione) restored wild-type response kinetics in cry1 and completely suppressed its long-hypocotyl phenotype in blue light. Co-treatment of cry1 seedlings with Ca-prohexadione plus GA4 completely reversed the effects of the inhibitor, restoring the long-hypocotyl phenotype typical of the mutant. Treatment of wild-type seedlings with GA4 was not sufficient to phenocopy cry1 seedlings, but co-treatment with IAA plus GA4 produced cry1-like growth kinetics for a period of approximately 5 h. The genomic and physiological data together indicate that blue light acting through cry1 quickly affects the expression of many genes, a subset of which suppresses stem growth by repressing GA and auxin levels and/or sensitivity. PMID- 14535886 TI - Increased tryptophan decarboxylase and monoamine oxidase activities induce Sekiguchi lesion formation in rice infected with Magnaporthe grisea. AB - Sekiguchi lesion (sl)-mutant rice infected with Magnaporthe grisea showed increased light-dependent tryptophan decarboxylase (TDC) and monoamine oxidase (MAO) activities. TDC and MAO activities were observed before the penetration of M. grisea to rice cells and maintained high levels even after Sekiguchi lesion formation. Light-dependent expression of TDC gene was observed in leaves inoculated with M. grisea before Sekiguchi lesion formation. Spore germination fluid (SGF) of M. grisea also induced Sekiguchi lesion formation accompanied by increased enzymes activities and tryptamine accumulation. Sekiguchi lesion was also induced by treatments with tryptamine and beta-phenylethylamine, which are substrates for MAO, but was not induced by non-substrates such as indole-3 propionic acid, (+/-)-phenylethylamine and tryptophan under light. Light dependent induction of Sekiguchi lesion by tryptamine was significantly inhibited in the presence of MAO inhibitors, metalaxyl and semicarbazide, and H2O2 scavengers, ascorbic acid and catalase. H2O2 in M. grisea-infected leaves with and without Sekiguchi lesions was demonstrated directly in situ by strong 3,3' diaminobenzidine (DAB) staining. On the other hand, H2O2 induced Sekiguchi lesions on leaves of cv. Sekiguchi-asahi under light, but not in darkness. This difference was associated with the decrease of catalase activity in infected leaves under light and the absence of decrease in darkness. We hypothesize that the H2O2-induced breakdown of cellular organelles such as chloroplasts and mitochondria in mesophyll cells may cause high TDC and MAO activities and the development of Sekiguchi lesion, and that the sl gene products in wild-type rice may function as a suppressor of organelle breakdown caused by chemical or environmental stress. PMID- 14535887 TI - Vacuolar cation/H+ exchange, ion homeostasis, and leaf development are altered in a T-DNA insertional mutant of AtNHX1, the Arabidopsis vacuolar Na+/H+ antiporter. AB - The function of vacuolar Na+/H+ antiporter(s) in plants has been studied primarily in the context of salinity tolerance. By facilitating the accumulation of Na+ away from the cytosol, plant cells can avert ion toxicity and also utilize vacuolar Na+ as osmoticum to maintain turgor. As many genes encoding these antiporters have been cloned from salt-sensitive plants, it is likely that they function in some capacity other than salinity tolerance. The wide expression pattern of Arabidopsis thaliana sodium proton exchanger 1 (AtNHX1) in this study supports this hypothesis. Here, we report the isolation of a T-DNA insertional mutant of AtNHX1, a vacuolar Na+/H+ antiporter in Arabidopsis. Vacuoles isolated from leaves of the nhx1 plants had a much lower Na+/H+ and K+/H+ exchange activity. nhx1 plants also showed an altered leaf development, with reduction in the frequency of large epidermal cells and a reduction in overall leaf area compared to wild-type plants. The overexpression of AtNHX1 in the nhx1 background complemented these phenotypes. In the presence of NaCl, nhx1 seedling establishment was impaired. These results place AtNHX1 as the dominant K+ and Na+/H+ antiporter in leaf vacuoles in Arabidopsis and also suggest that its contribution to ion homeostasis is important for not only salinity tolerance but development as well. PMID- 14535888 TI - Stress memory in plants: a negative regulation of stomatal response and transient induction of rd22 gene to light in abscisic acid-entrained Arabidopsis plants. AB - All organisms, including plants, perceive environmental stress, and they use this information to modify their behavior or development. Here, we demonstrate that Arabidopsis plants have memory functions related to repeated exposure to stressful concentrations of the phytohormone abscisic acid (ABA), which acts as a chemical signal. Repeated exposure of plants to ABA (40 micro m for 2 h) impaired light-induced stomatal opening or inhibited the response to a light stimulus after ABA-entrainment under both dark/light cycle and continuous light. Moreover, there were transient expressions of the rd22 gene during the same periods under both the growing conditions. Such acquired information in ABA-entrained plants produced a long-term sensitization. When the time of light application was changed, a transient induction of the rd22 gene in plants after ABA-entrainment indicated that these were light-associated responses. These transient effects were also observed in kin1, rab18, and rd29B. The transient expression of AtNCED3, causing the accumulation of endogenous ABA, indicated a possible regulation by ABA-dependent pathways in ABA-entrained plants. An ABA immunoassay supported this hypothesis: ABA-entrained plants showed a transient increase in endogenous ABA level from 220 to 250 pmol g-1 fresh mass at 1-2 h of the training period, whereas ABA-deficient (aba2) mutants did not. Taking into account these results, we propose that plants have the ability to memorize stressful environmental experiences, and discuss the molecular events in ABA-entrained plants. PMID- 14535889 TI - AtAGP30, an arabinogalactan-protein in the cell walls of the primary root, plays a role in root regeneration and seed germination. AB - Arabinogalactan-proteins (AGPs) are extracellular proteoglycans that are implicated in many plant growth and developmental processes, but in no case has a biological function been assigned to a particular AGP. AtAGP30 is a non-classical AGP core protein from Arabidopsis that is expressed only in roots. Analysis of the corresponding mutant, agp30, has revealed that the wild-type gene product is required in vitro for root regeneration and in planta for the timing of seed germination. The mutant shows a suppression of the abscisic acid (ABA)-induced delay in germination and altered expression of some ABA-regulated genes. This suggests that AtAGP30 functions in the ABA response. By analogy to proteoglycan mediated regulation of growth-factor-signalling pathways in animals, our data indicate that phytohormone activity in plants can be modulated by AGPs. PMID- 14535890 TI - Telomere looping in P. sativum (common garden pea). AB - Telomeres vary greatly in size among plants and, in most higher plants, consist of a long array of 5'-TTTAGGG-3'/3'-AAATCCC-5' (TTTAGGG) repeats. Recently, telomeric DNA in human, mouse, oxytricha, and trypanosome chromosomes have been found arranged into loops (t-loops), proposed to sequester the telomere from unwanted repair events and prevent activation of DNA damage checkpoints. We have asked whether t-loops exist in the higher order plant Pisum sativum (garden pea). DNA was isolated from the shoots and root tips of germinating seeds. Analysis of the telomeric restriction fragments showed that DNA hybridizing to a (TTTAGGG)n probe migrated as a smear centering around 25 kb, and direct sequencing verified the repeat to be (TTTAGGG)n. Total DNA in isolated nuclei was photo-cross-linked, and the telomeric restriction fragments were purified by gel filtration. Electron microscopic (EM) analysis revealed DNA molecules arranged as t-loops with a size distribution consistent with that seen by gel electrophoresis. Some molecules had loops as large as 75 kb. These results show that the arrangement of telomeric DNA into loops occurs in higher plants. PMID- 14535891 TI - Cell damage and reactive oxygen species production induced by fluorescence microscopy: effect on mitosis and guidelines for non-invasive fluorescence microscopy. AB - The green fluorescent protein (GFP) and other intrinsically fluorescent proteins (IFPs) are popular reporters because they allow visualization of cellular constituents in living specimens. IFP technology makes it possible to view dynamic processes in living cells, but extended observation, using fluorescence microscopy (both wide-field and confocal), can result in significant light energy exposure. Therefore, it is possible that cells experience light-induced damage that alters cell physiology and confounds observations. To understand the impact that extended viewing has on cells, we obtained quantitative information about the effect of light energy dose and observation conditions on tobacco BY-2 cell physiology. Our results show a non-linear relationship between the excitation light intensity and mitotic arrest, and the frequency of mitotic arrest is dependent on the presence of an IFP that absorbs the excitation light. Moreover, fluorescence microscopy induces the production of reactive oxygen species (ROS), as assayed using BY-2 cells loaded with oxidation-sensitive dyes, and the level of ROS production increases if the cells express an IFP that absorbs the excitation light energy. The dye oxidation follows sigmoidal kinetics and is reversible if the cells are exposed to low irradiation levels. In addition, the dye oxidation rate shows a non-linear relationship to the excitation light intensity, and a good correlation exists between photobleaching, mitotic arrest, and dye oxidation. The data highlight the importance of ROS scavenging for normal mitotic progression, and provide a reference for judiciously choosing conditions that avoid photobleaching that can lead to ROS accumulation and physiological damage. PMID- 14535892 TI - Treatment of polymorphic light eruption. AB - Polymorphic light eruption (PLE) is a highly prevalent photosensitivity disorder, estimated to affect 11-21% people in temperate countries. Typically, PLE appears as a recurrent pruritic eruption comprising papules and/or vesicles and/or plaques, which occurs on photo-exposed skin sites following sun exposure, and which heals without scarring. Commoner in females, the aetiology is uncertain, although there is evidence of an immune basis. We perform a review of the prophylaxis and treatment of this condition. While sun protection, corticosteroids and desensitization phototherapy are the mainstays of management, a range of anti-inflammatory and immunomodulatory agents are reported. PMID- 14535893 TI - Cutaneous effects of infrared radiation: from clinical observations to molecular response mechanisms. AB - Human skin is exposed to infrared (IR) radiation (760 nm-1 mm) from natural as well as artificial sources that are increasingly used for cosmetic or medical purposes. Epidemiological data and clinical observations, however, indicate that IR radiation cannot be considered as totally innocuous to human skin. In particular, IR radiation, similar to ultraviolet radiation, seems to be involved in photoaging and potentially also in photocarcinogenesis. The molecular consequences resulting from IR exposure are virtually unknown. Recent studies, however, have begun to shed light on the basic molecular processes such as cellular signal transduction and gene expression triggered by exposure to IR radiation. In response to IR irradiation, mitogen-activated protein kinase signaling pathways were activated mediating the upregulation of matrix metalloproteinase-1 expression. This previously unrecognized molecular 'IR response' shows that IR radiation is capable of specifically interfering with cellular functions and provides a molecular basis for biological effects of IR on human skin. PMID- 14535894 TI - Dietary supplementation of (+)-catechin protects against UVB-induced skin damage by modulating antioxidant enzyme activities. AB - PURPOSE: The aim of this study was to investigate the effects of dietary supplementation with (+)-catechin on cutaneous antioxidant enzymes and the skin damage caused by UVB irradiation. METHODS: BALB/c mice were divided into three groups. Each group was fed a regular diet (RD) or a 2% catechin-supplemented diet for either 2 weeks (2CSD) or 4 weeks (4CSD) ad libitum prior to UVB irradiation. Skin was removed for the antioxidant enzyme assay, hematoxylin and eosin staining, and the TEM analysis before and at various time points after UVB (200 mJ/cm2) irradiation. RESULTS: Before UVB irradiation, the superoxide dismutase (SOD) and catalase (CAT) activities of the 2CSD and the 4CSD groups were found to be lower than those of the RD group, whereas the glutathione peroxidase (GPx) activity of the 4CSD group was higher than those of the RD and the 2CSD groups (P<0.05). The SOD and CAT activities of the RD group decreased after UVB irradiation, while those of the 2CSD and the 4CSD groups increased immediately after irradiation and then decreased (P<0.05). Immediately after UVB irradiation, the GPx activities of the 4CSD and the 2CSD groups increased, but that of the RD group decreased. The GPx activity of all three groups showed a tendency to return to pre-UVB irradiation levels with time. Light microscopic findings of the RD group showed epidermal thinning and apoptotic cells at 24 h after UVB irradiation and mostly necrotic cells at 48 h, whereas only moderate thickening of the epidermis was observed in the 2CSD group at 48 h after irradiation. An electron microscopic examination produced similar findings. At 48 h after irradiation, nearly all epidermal cells seemed to be damaged in the RD group as compared to the 2CSD group. CONCLUSION: These results demonstrate that dietary supplementation with (+)-catechin could protect epidermal cells against UVB induced damage by modulating antioxidant enzyme activities. PMID- 14535896 TI - Simplified method to substantiate SPF labeling for sunscreen products. AB - BACKGROUND/PURPOSE: Worldwide, sunscreen sun protection factor (SPF) testing is based on 30-year-old technology. At the time the SPF test came into being, the highest SPFs available were in the 6-8 range. The SPF test is reasonably accurate for SPFs up to 15, but is much less reliable for measuring SPFs of 30 and higher. The method we propose addresses two primary reasons for this unreliability: 1) difficulties in applying products uniformly and 2) the subjectivity and variability of perception in evaluating and grading responses to UV doses. METHODS: Our proposed SPF substantiation method differs from the current SPF test in that sunscreen-protected test sites receive the same UV dose in four uniformly spaced sub-sites, which are graded as passing if no response is seen or failing if any response is seen. The response may be tanning, erythema, or a combination of both. To demonstrate the method, two commercial products with labeled SPFs of 30 and 45 and the P2 sunscreen standard were tested at two different laboratories. RESULTS: The SPF 30 product and SPF 15 standard were shown to be correctly labeled. However, it is questionable as to whether the SPF 45 product provides protection against 45 minimal erythema doses. CONCLUSIONS: Our proposed SPF substantiation method is not dependent on subjective evaluation of responses, accounts for non-uniform product application, and provides a conservative estimate of sunscreen protection. The method consists of a systematic repetition of identical tests that are considerably more rigorous than the current methods that are based on single data points per test subject. While the current SPF test remains necessary and valuable as a dose ranging tool, we propose that this SPF substantiation method supersede the old method for final SPF label determination. PMID- 14535895 TI - Assessment of the skin photoprotective capacities of an organo-mineral broad spectrum sunblock on two ex vivo skin models. AB - UV irradiation can cause cutaneous damage that may be specific according to the wavelength of UV rays. For example, damage from UVB irradiation manifests itself in the form of sunburn cells and enhancement of the expression of p53, while damage from UVA exposure results in an increase in the expression of vimentin. These reactions to UV irradiation were used in this work to evaluate the photoprotective capacities of two sunblock preparations that were applied to the surface of the skin. One sunblock preparation is a UVB absorber containing zinc oxide (ZnO) and titanium oxide (TiO2) exclusively. The other sunblock preparation is a new organo-mineral sunblock containing Tinosorb M, OCM, ZnO and TiO2. Evaluation of the photoprotective capacities of both preparations on hairless rat skin and on in vitro reconstructed human epidermis revealed that they were effective in preventing UVB-induced damage. In contrast, only the organo-mineral sunblock was effective in the prevention of UVA-specific damage such as dermal alterations characterized by the expression of vimentin. Furthermore, our data support the fact that hairless rat skin and in vitro reconstructed human epidermis are a reliable basis for the evaluation of the photoprotective capacities of various sunscreens against UVB and UVA damage. PMID- 14535897 TI - Assessment of the usefulness of skin phototype and skin color as the parameter of cutaneous narrow band UVB sensitivity in psoriasis patients. AB - BACKGROUND: Many reports have been released to assess skin types, skin colors and cutaneous sensitivity to broad band UVB or UVA. OBJECTIVE: This study was performed to investigate the usefulness of skin type and skin color as the parameter of narrow band UVB (NBUVB) sensitivity. METHODS: The minimal erythema dose (MED) of 40 psoriasis patients was investigated by irradiating several doses ranging from 200 to 1500 mJ/cm2. Before phototesting, the skin color of buttock was measured with a tristimulus colorimeter. RESULTS: The median and mode value of MED of NBUVB was 950 mJ/cm2. Skin type was well correlated with the MED and there was a significant relationship between the L* value and MED, but not for the a* and b* values. CONCLUSION: The MED value of NBUVB in our study is a basic data to set the phototherapy protocol. Our result showed that skin type and L* value might be useful for predicting the sensitivity to NBUVB irradiation. PMID- 14535898 TI - Narrowband UVB therapy in the treatment of lichen planus. AB - BACKGROUND: Recently, UVB lamps with a peak emission around 311 nm have been used successfully for the treatment of many dermatologic diseases known to be treated with psoralen plus UVA (PUVA). Narrowband UVB (NBUVB) radiation causes less erythema and carcinogenicity with lower cumulative doses than PUVA, while the treatment response remains high. Lichen planus (LP) is a cell-mediated immune response of unknown origin. METHODS: We present our results of NBUVB therapy administered to 10 LP patients. The sessions were administered three to four times weekly with an average cumulative dose of 17.7+/-1.6 J/cm2. RESULTS: Five patients responded completely, and four were partially responsive at the end of the 30th session. Three of the partially responsive cases responded completely at the 31st, 36th and 51st sessions, respectively. CONCLUSION: Clinical improvements observed in our study as well as the potential advantages of NBUVB imply that it is an inevitable treatment alternative for resistant cases of LP. PMID- 14535899 TI - Reply to sunbathing versus indoor tanning: a realistic perspective. PMID- 14535900 TI - Bereavement in paediatric intensive care. AB - The death of a child is a very sad event in anyone's life. It also affects all staff in paediatric intensive care units in different ways at different times. The publication of standards of bereavement care in paediatric intensive care hopes to be able to assist medical and nursing staff to understand and feel more confident in this emotionally difficult area of medicine. The aim of this article is to summarize some of the major points made in the document. PMID- 14535901 TI - Chemokines and the inflammatory response following cardiopulmonary bypass--a new target for therapeutic intervention?--A review. AB - This 10-year Medline search of English-language articles describing experimental and clinical studies on chemokines, cardiopulmonary bypass (CPB) and systemic or multiorgan failure revealed that chemokines are significantly involved in the pathogenesis of post-CPB syndrome. The post-CPB inflammatory response depends upon recruitment and activation of inflammatory cells. Leucocyte recruitment is a well-orchestrated process that involves several protein families, including pro inflammatory cytokines, adhesion molecules and chemokines. Current anti inflammatory therapies mostly act on the cells that have already been recruited. A more efficient therapy might be the prevention of excessive recruitment of particular leucocyte populations by antagonizing chemokine receptors which might act upstream of the current anti-inflammatory agents. The chemokines, which are a cytokine subfamily of chemotactic cytokines, participate in recognizing, recruiting, removing and repairing inflammation. As chemokines target specific leucocyte subsets, antagonism of a single chemokine ligand or receptor would be expected to have a circumscribed effect, thereby endowing the antagonist with a limited side-effect profile. Chemokines should be considered as possible targets for therapeutic intervention. PMID- 14535902 TI - Oral high-dose midazolam premedication for infants and children undergoing cardiovascular surgery. AB - BACKGROUND: The purpose of this study was to determine whether oral midazolam 1.5 mg x kg(-1) is a safe and effective alternative to standard-dose midazolam (0.5 1.0 mg x kg(-1)) premedication for infants and children with congenital heart disease. METHODS: A total of 193 infants and children (4 months to 2 years) undergoing cardiovascular surgery were studied. Each patient received 0.5, 1.0, or 1.5 mg x kg(-1) of oral midazolam. The level of sedation was assessed with a 5 point scale and vital signs were measured including blood pressure (BP), heart rate (HR) and oxyhaemoglobin saturation (SpO2) before and after the medication. RESULTS: Infants and children premedicated with oral midazolam 1.5 mg x kg(-1) were better sedated than those with standard-dose midazolam: 4% of infants and children given 1.5 mg x kg(-1) of midazolam became agitated compared with 14% given 1.0 mg x kg(-1) and 26% in those given 0.5 mg x kg(-1). Ninety percentage of infants and children given 1.5 mg x kg(-1) of midazolam achieved satisfactory sedation (calm, drowsy, or asleep) in 30 min, whereas 68% in those given 1.0 mg x kg(-1) and 35% in those given 0.5 mg x kg(-1). Midazolam 1.5 mg x kg(-1) did not cause any statistically significant decrease in BP, HR, or SpO2, although eight infants and children showed > or =20% drop in systolic BP and six infants and children showed >5% drop in SpO2. No 'spelling attacks', seizure-like activity, apnoea, nor laryngospasm were observed in any infants and children during and after the medication. CONCLUSIONS: Oral midazolam 1.5 mg x kg(-1) is excellent for preanaesthetic medication for infants and children undergoing cardiovascular surgery. PMID- 14535903 TI - Latex allergy in children with oesophageal atresia. AB - BACKGROUND: Latex allergy is frequently found in children and patients with spina bifida and urogenital abnormalities and have been considered at risk for latex sensitization. The aim of the study was to evaluate the incidence of latex sensitization in patients with oesophageal atresia and undergoing three or more surgical procedures and to identify possible risk factors in the process of latex sensitization. METHODS: A total of 20 patients were analysed: 19 boys and one girl. The oesophageal atresias were as follows: type I in three children, type II in two and type III in 15 children. Surgical and anaesthetic procedures, intensive care management, age, type of oesophageal atresia, associated congenital malformations, Waterston and Montreal prognostic classifications were considered as risk factors that may be implicated in the process of sensitization. RESULTS: Five patients (25%) were considered sensitized to latex (group 1) and 15 (75%) nonsensitized (group 2). Among the five sensitized patients, three reported clinical reactions to latex, while the other two presented only specific IgE sensitization. The number of operations, the total hours of surgery, the number of drainages, the total days of drainage, the total days of central venous catheter were significantly greater in group 1 than in group 2. Both of the highest risk oesophageal atresia classes (Waterston C and Montreal II) were related to latex allergy. CONCLUSIONS: Oesophageal atresia, especially in cases of prolonged management, must be considered as a risk for the development of latex allergy. PMID- 14535904 TI - Investigation of the radiological relationship between iliac crests, conus medullaris and vertebral level in children. AB - BACKGROUND: The inability of anaesthetists to accurately identify vertebral spaces has been documented. The aim of our magnetic resonance imaging (MRI) study was to find the relationship of vertebral body level with Tuffier's line and the conus medullaris in children. METHODS: Forty-nine children aged 10 years or less who had MRI scanning in the region of the lumbar spine were identified. If either the conus medullaris or iliac crests were visible, the corresponding vertebral body level was recorded. RESULTS: The level of the conus could be identified in 43 patients. The median conus level was at L1 with an interquartile range (IQR) of 0.5 vertebral levels. The level of the crests could be identified in 35 patients. The median crest height was L5 with an IQR of 0.5 vertebral levels. The difference between conus and crest levels could only be calculated in 29 patients. In the remainder of the children one of the two pieces of the data was not available. The median difference was 4 vertebral levels with an interquartile range (IQR) of 1. Two of 49 children, both of whom had tethered cords, had differences of <2 vertebral levels. CONCLUSIONS: Our data confirmed that the iliac crests and conus medullaris were consistently related to the predicted vertebral levels in children. In the absence of a tethered spinal cord, subarachnoid puncture at L4-5 or below should present little risk of conus damage. Tuffier's line corresponded with a radiological level of L4-L5 in our study population. PMID- 14535905 TI - Detection of epidural catheters with ultrasound in children. AB - BACKGROUND: The aim of this study was to assess whether a noninvasive imaging technique such as ultrasound could visualize an epidural catheter in the epidural space in children. METHODS: Following local ethics committee approval and informed parental consent a pilot study of 12 cases was performed. Children undergoing major surgery requiring epidural analgesia were recruited. All catheters were introduced via the lumbar region. All children were scanned within 24 h of epidural insertion by consultant paediatric radiologists. If the catheter was identified in the epidural space then an attempt was made to visualize the entire length of the catheter. RESULTS: The epidural catheter was detected in nine of 12 patients. All of these were less than 6 months old. The entire length of the catheter was visualized in five of the nine patients. It was possible to estimate the most cephalad level of the catheter in seven of the nine patients. This was in the thoracic region in all cases and an appropriate level for the intended surgical procedure. It was not possible to precisely identify the tip of the catheter as a distinct entity using ultrasound. CONCLUSION: This study shows that it is possible to visualize an epidural catheter in the epidural space in children under 6 months of age using ultrasound. PMID- 14535906 TI - Laryngeal mask insertion in children: a rational approach. AB - BACKGROUND: Various techniques of laryngeal mask airway (LMATM) insertion have been described in adults but only limited clinical trials have been conducted in children despite a varying range in success rate by the recommended method. METHOD: The study was conducted in 62 ASA 1 and 2 children who were randomly allocated for the evaluation of LMA insertion by the midline approach with the cuff completely deflated (group MD, control group, n = 31) or laterally with the cuff partially inflated (group LP, study group, n = 31). Propofol was used as the sole induction agent in all children. Ease of insertion, position of the LMA with fibreoptic laryngoscope and incidence of stomach insufflation were assessed. RESULTS: Fewer attempts and a significant reduction in the time for insertion was noted in group LP (14.4 +/- 4.2 s) compared with group MD (23.1 +/- 2.1 s), P < 0.05. Despite a good seal around the cuff and satisfactory ventilation a significantly higher incidence of malposition of the LMA was recorded by intraluminal fibreoptic endoscopy in group MD (13% to nil), P < 0.05. Similarly gastric insufflation was significantly greater in group MD (42% compared with 10%). In children with grade 3 fibreoptic view significantly higher endtidal carbon dioxide values were recorded throughout the study period after LMA insertion until its removal. LMA was stained with blood in 13% children in group MD compared with 3% in group LP at the time of removal. CONCLUSION: A partially inflated cuff inserted by the lateral route is a better method of insertion in children and grade 3 fibreoptic views can be associated with a significant build up of carbon dioxide in children breathing spontaneously. PMID- 14535907 TI - Laryngeal mask airway for ventilation during diagnostic and interventional fibreoptic bronchoscopy in children. AB - BACKGROUND: The use of the flexible fibreoptic bronchoscope in infants and children has expanded to include both interventional and diagnostic bronchoscopy. The present report utilizes the laryngeal mask airway (LMATM) for ventilation and anaesthesia administration in children during fibreoptic bronchoscopy using an adult bronchoscope. METHODS: The technique was used in 10 children; their age ranged between 1.2 and 5 years. Six of these children had a history of foreign body aspiration and underwent interventional bronchoscopy, while four children underwent diagnostic bronchoscopy. Anaesthesia was induced by facemask using sevoflurane 6-8% followed by the insertion of a LMA size 2 or 2.5. A swivel adapter connected the proximal end of the LMA to the T-piece anaesthesia system. Anaesthesia was then maintained with spontaneous breathing of sevoflurane 2-4% in oxygen, delivered via the LMA. A flexible adult fibreoptic bronchoscope (outer diameter 4.9 mm) was inserted via the swivel adapter. RESULTS: The procedure could be accomplished successfully in the 10 patients. However, one child developed laryngospasm that was easily relieved by deepening the level of anaesthesia. In a second child hypoxia and hypercarbia occurred and were relieved by intermittent withdrawal of the fibreoptic bronchoscope. CONCLUSIONS: Laryngeal mask airway is a safe and effective adjunct to fibreoptic bronchoscopy under general anaesthesia in children. Its larger internal diameter compared with a tracheal tube permits the use of relatively large fibreoptic bronchoscope without a significant increase in airway resistance. PMID- 14535908 TI - Remifentanil vs alfentanil in the total intravenous anaesthesia for paediatric abdominal surgery. AB - BACKGROUND: Our aim was to investigate whether total intravenous anaesthesia (TIVA) with remifentanil and alfentanil would ensure appropriate analgesia and recovery conditions in anaesthesia for children undergoing abdominal surgery. METHODS: Sixty children, scheduled for abdominal operations were randomized to receive, in a double-blind manner, either remifentanil (loading dose 1 microg x kg(-1); maintenance infusion, 0.25 microg x kg(-1) min(-1)) or alfentanil (loading dose 50 microg x kg(-1); maintenance infusion, 1 microg x kg(-1) min( 1)) as the analgesic component of TIVA. They were combined with propofol (loading dose, 2 mg x kg(-1); step 1 maintenance infusion, 10 mg x kg(-1) h(-1); step 2 maintenance infusion, 8 mg x kg(-1) h(-1); step 3 maintenance infusion, 6 mg x kg(-1) h(-1)) neuromuscular blockade was with mivacurium. Dose changes of the drugs, the times from cessation of anaesthesia to extubation, verbal responses, recovery of ventilation, orientation, and qualification for discharge from the postanaesthetic care unit (PACU) were recorded. RESULTS: Demographics, duration of surgery and anaesthesia were similar between the two groups. Times to extubation and stay in the PACU were significantly shorter in the remifentanil group compared with the alfentanil group. Quality of emergence (QE) from anaesthesia scale scores were higher in the remifentanil group compared with the alfentanil group. CONCLUSIONS: Remifentanil provides a more rapid recovery and adequate postoperative analgesia after TIVA for paediatric abdominal surgery, compared with alfentanil. PMID- 14535909 TI - Remifentanil infusion for cleft palate surgery in young infants. AB - BACKGROUND: The residual depressant effect of opioid is a major concern in infants scheduled for cleft palate repair. Remifentanil is associated with a fast and predictable recovery, independent of age. METHODS: About 40 infants in the 2 12 month age range were prospectively enrolled in this open study, to receive either remifentanil (infusion starting at 0.25 microg x kg(-1) x min(-1)) or sufentanil as part of a balanced anaesthesia regimen. Isoflurane was maintained at an endtidal concentration of 1.2% in oxygen and nitrous oxide and the opioid dosing was titrated to autonomic responses. Postoperative pain relief was provided by morphine infusion. Morphine administration started intraoperatively in the remifentanil group. RESULTS: Consistent haemodynamic stability was achieved throughout surgery in both groups. Infants of the remifentanil group required, on average, lower concentrations of isoflurane than children of the sufentanil group (1.2 +/- 0.2% vs 1.7 +/- 0.3%, P < 0.001). The median time from last suture to tracheal extubation was 12.5 min (5-25 min) in the remifentanil group and 15.0 min (10-30 min) in the sufentanil group. There was no evidence of hyperalgesia or enhanced morphine consumption in the remifentanil group compared with the sufentanil group. Postoperative pain scores were even lower in the remifentanil group, compared with the sufentanil group, soon after arrival in the postanaesthesia care unit. CONCLUSIONS: Remifentanil-based anaesthesia appeared well suited for primary cleft palate repair in young infants. PMID- 14535910 TI - Validation of a six-graded faces scale for evaluation of postoperative pain in children. AB - BACKGROUND: The faces pain scales are often used for self-report assessment of paediatric pain. The aim of this study was to evaluate the validity of a six graded faces pain scale after surgery by comparing the level of agreement between the children's report of faces pain scores and experienced nurses' assessment of pain by observation of behaviour. The faces pain scores before, at and after administration of analgesics were analysed. The study was performed in two South African hospitals, one with a mainly rural population and the other with an urban population. METHODS: A total of 110 children aged 4-12 years, scheduled for inguinal surgery in the two South African hospitals, were included in the study. The anaesthetic technique was standardized. All patients received a caudal block preoperatively. Postoperative pain assessments were made every hour for 8 h after the caudal block was performed. A designated nurse assessed pain by using a four graded descriptive scale (no, mild, moderate or severe pain) and thereafter the child reported pain by using the six-graded faces pain scale. RESULTS: A high correlation was found between the two methods of assessment (tau = 0.76, P < 0.0001). The correlation between methods was high in both hospital populations and in all age groups. The weakest correlation was found in children aged 8-12 years (tau = 0.56, P < 0.01). Significantly lower faces pain scores were found after administration of analgesics compared with pain rating before analgesics (P < 0.0001). The proportion of patients with pain scores above 2 decreased from 86% to 31% (P < 0.001). CONCLUSIONS: The findings support this six-graded faces pain scale as a useful and valid instrument for measuring pain in the postoperative period in children aged 4-12 years. PMID- 14535911 TI - General anaesthesia for lobectomy in an 8-year-old child with Kartagener's syndrome. AB - Kartagener's syndrome (KGS) is an autosomal recessive disorder characterized by the clinical triad of bronchiectasis, sinusitis and dextrocardia (situs inversus). Reports of general anaesthesia in a child with KGS are rare in medical literature. We describe the case of a young boy who underwent lobectomy under general anaesthesia using a single lumen tube. Anaesthetic implications of this syndrome are mentioned. Key points in the management of anaesthesia in KGS are discussed. PMID- 14535912 TI - A long-term continuous infusion via a sciatic catheter in a 3-year-old boy. AB - We describe the case of a 3-year-old boy with a subtotal amputation of the right foot who received treatment for pain via a peripheral catheter positioned at the level of the sciatic nerve (lateral approach).We administered a continuous infusion of 0.2% ropivacaine, 0.4 mg x kg(-1) x h(-1) plus clonidine 0.12 microg x kg(-1) x h(-1) for 21 days. Pain relief was complete and the patient did not require any further rescue analgesia throughout the period even during medications and surgical treatment in our intensive care unit. We discuss the safety and efficacy of the use of a peripheral continuous infusion in children compared with other techniques of analgesia. PMID- 14535913 TI - A rare cause of upper airway obstruction in a 5-year-old girl: a laryngeal web. AB - A case is described of a 5-year-old girl with respiratory distress. She was intubated without difficulty, but developed respiratory distress on extubation. Laryngoscopy/bronchoscopy via a laryngeal mask airway revealed an extensive layngeal web which was removed with laser therapy. The causes and differential diagnosis of laryngeal web are discussed. PMID- 14535914 TI - Anaesthetic implications of Robinow syndrome. AB - Robinow (fetal face) syndrome is a rare inherited multisystem disorder featuring mesomelic or acromesomelic limb shortening, facial and spinal deformities, hypoplastic genitalia, kidney disease and congenital heart defects. We report the anaesthetic management of a patient with Robinow syndrome presenting for elective surgery and review specific issues of interest in the perioperative workup. PMID- 14535915 TI - Systemic embolism in an infant following haemangioma embolization: a two-step process. AB - A case is presented of therapeutic embolization of a hypervascular hamartoma of the liver, in a term baby. During the procedure signs of pulmonary embolism occurred and the baby subsequently died from myocardial ischaemia. Potential intrathoracic shunts of the newborn together with changes associated with the vascular tumour are thought to have allowed systemic embolization of the embolic material. PMID- 14535916 TI - Anaesthetic management in children with metachromatic leukodystrophy. AB - We present a case of metachromatic leukodystrophy in a child who required surgery for gastro-oesophageal reflux. In spite of his demyelinating disease, we used a lumbar epidural technique with general anaesthesia; the epidural catheter allowed us to continue the analgesia postoperatively and to avoid opioids in this high risk patient. PMID- 14535917 TI - Intraoperative transoesophageal echocardiography in a low birth weight neonate with atrioventricular septal defect. AB - An 18-day-old male neonate (45 cm, 1.8 kg) with a history of cyanosis and congestive heart failure from an atrioventricular septal defect (AVSD) with a large left-to-right shunt was scheduled for surgical repair of the AVSD. After routine induction of anaesthesia with fentanyl and vecuronium, a 4.5-mm diameter transoesophageal echocardiography (TOE) probe was inserted into the oesophagus, and systematic echocardiographic evaluation was performed during surgery. After cardiopulmonary bypass was stopped, intraoperative TOE revealed mild residual mitral valve regurgitation. Because good left ventricular wall motion was confirmed and haemodynamic parameters were stable, cardiopulmonary bypass was not reinitiated. The patient's cardiac output was low in the postoperative intensive care unit. TOE was performed the next day to detect the source of this problem, revealed severe regurgitation compared with that observed intraoperatively. TOE was useful for evaluation of the residual mitral valve regurgitation, and we reconfirmed the importance of continuous monitoring even in a low birthweight neonate undergoing repair of a complete AVSD. PMID- 14535918 TI - Defective endotracheal tube--cause of airway obstruction. PMID- 14535919 TI - Anaesthetic management of a strabismus patient with phenylketonuria. PMID- 14535920 TI - Postoperative apnoea in an ex-premature infant: is it only related to clonidine? PMID- 14535922 TI - Postintubation tracheal stenosis in an 11-year-old boy: a surgical and anaesthetic challenge. PMID- 14535924 TI - A lack of a proper pharmaceutical presentation of vecuronium used in paediatric anaesthesia may affect drug utilization and costs. PMID- 14535925 TI - Airway management without i.v. access-- bad practice or a technique with potential? PMID- 14535926 TI - Instrumentation of the airway in the absence of intravenous access. PMID- 14535927 TI - Positive identification of Newcastle disease virus vaccine strains and detection of contamination in vaccine batches by restriction site analysis of the matrix protein gene. AB - Twelve vaccine batches prepared from avirulent vaccine strains of Newcastle disease virus produced by seven manufacturers were identified by analysis of the matrix (M) protein gene with restriction enzymes MboI and HinfI. The analyses have revealed the presence of the strain indicated by the manufacturers (namely B 1, LaSota or Ulster 2C), except in one case when the vaccine contained strain V4 Queensland instead of VGGA as indicated. In addition, several batches of both monovalent and combined vaccines containing strain LaSota of the same company consistently disclosed contamination with strain B-1. The mixed nature of the preparations was verified not only by the dual patterns of restriction fragments but also by separating the two components and identifying them individually. Restriction analysis of the M gene, by allowing positive identification of each of the lentogenic vaccine strains, should provide an improvement in controlling vaccine batches by revealing homologous contaminants or exchange of the vaccine strain. PMID- 14535928 TI - Studies on the safety and immunogenicity of the South African bluetongue virus serotype 2 monovalent vaccine: specific detection of the vaccine strain genome by RT-PCR. AB - In order to study the safety and the immunogenicity of the South African vaccine against the serotype 2 bluetongue virus, two groups of seven sheep were vaccinated with the vaccine used in the French island of Corsica. Vaccinated and non-vaccinated sheep were observed clinically and their rectal temperatures were recorded daily. The serological response in vaccinated animals confirmed the immunogenicity of the vaccine. Post-vaccinal viraemia was investigated and the vaccine genome was detected by reverse transcription polymerase chain reaction (RT-PCR). No viraemia was observed at post-vaccination days 4, 7 and 11 but the vaccine strain of virus was detected by RT-PCR throughout the experiment. The thermostability of the vaccine was also evaluated. The vaccine titre strongly decreased at temperatures higher than 35 degrees C. PMID- 14535929 TI - Spatial relative risk mapping of pseudorabies-seropositive pig herds in an animal dense region. AB - This paper describes a new approach for spatial relative risk mapping as a tool for geographical risk assessment. The spatial epidemiological analysis is based on geographically referenced data about pseudorabies (Aujeszky's disease) virus infections at farm level in a region of high animal density in Germany at the beginning of the national eradication project. On the basis of serological findings 186 farms were classified as positive out of a total of 482 investigated farms listed in veterinary administrative registers. Geographical cluster analysis was used to identify two areas of high risk (RR = 2.4 and 3.3). Non parametric density estimation was used to estimate the proportion of infected farms per square kilometre. Furthermore, the spatial relative risk function was approximated through the prevalence ratio defined by the ratio of the local prevalence and the overall prevalence of the farms outside the cluster regions. The corresponding approximated relative risk map indicates and quantifies a clear spatial pattern of disease frequency. PMID- 14535931 TI - Diagnosis of porcine proliferative enteropathy: detection of Lawsonia intracellularis by pathological examinations, polymerase chain reaction and cell culture inoculation. AB - A total of 21 pigs aged 7-17 weeks with clinical symptoms suggestive for Porcine Proliferative Enteropathy were examined for Lawsonia intracellularis by analysing the following parameters: (i) intestinal gross and histological lesions, (ii) presence of comma-shaped bacteria in enterocytes by Warthin-Starry and a modified Ziehl-Neelsen stain, (iii) PCR amplification of L. intracellularis DNA from intestinal mucosa by using two oligonucleotide primer pairs targeting a 255-bp DNA fragment of the 16S rDNA-gene and a 319-bp DNA fragment of the L. intracellularis chromosome. Specificity of PCR reactions was confirmed by using DNA extracted from the L. intracellularis reference strain N343 (ATCC 55672) as well as by DNA sequence comparisons of PCR amplification products with data bank entries. Intestinal gross lesion indicative for PPE were observed in 20 pigs (95.2%). For all 21 pigs, the L. intracellularis aetiology was confirmed by histological as well as bacterioscopical examinations. Specific PCR amplification products were obtained from 20 pigs (95.2%). Taking PCR positivity as the definite criterion, L. intracellularis was diagnosed in 20 pigs from 11 herds in seven Swiss cantons (Argovia, Berne, Fribourg, Grisons, Lucerne, Schwyz, Thurgovia). To grow L. intracellularisin vitro, the cell culture method of Lawson et al. (J. Clin. Microbiol. 1993: 31, 1136-1142) was adopted. Inocula prepared from heavily infected fresh and frozen ileal mucosa of 15 pigs were cultured in rat enterocytic IEC-18 cells (ATCC CRL 1589). Six cell culture passages of 10 days each were completed. The reference strain N343 was examined for cultivability, accordingly. Except for occasional specific PCR amplifications from cell cultures up to the second passage, any indications for growth of L. intracellularis in IEC-18 cells were not found. PMID- 14535930 TI - Chromium supplementation enhances antibody response to vaccination with tetanus toxoid in cattle. AB - Nineteen multiparous late-pregnant dairy cows were divided into an experimental group (n = 10) and a control group (n = 9). Animals in the experimental group were fed a diet supplemented with chromium chelate admixed into wheat meal at 5 mg per animal per day. The supplementation was started 4 weeks before calving and stopped by the end of week 3 after calving. All the cows were vaccinated with tetanus toxoid in the fifth and sixth week of lactation. Monitoring of the characteristics of non-specific immunity did not show any significant between group differences in total and differential leucocyte counts, percentages of lymphocyte subpopulations, activities of lectin-stimulated lymphocytes, phagocytic activities, and the contents of total immunoglobulins and lysozyme in blood sera. Tetanus toxoid-specific antibody titres, those of the IgG2 isotype in particular, were higher in the experimental group than in the control animals. These results indicate that chromium supplementation at the level used in this experiment modulated the regulation of functions of the immune system. PMID- 14535932 TI - Isolation and antimicrobial susceptibility of Aeromonas salmonicida in rainbow trout (Oncorhynchus mykiss) in turkey hatchery farms. AB - Three Aeromonas salmonicida strains were isolated from the livers of 265 rainbow trouts sampled. The antibiotic susceptibility test results showed that A. salmonicida strains were susceptible to streptomycine and ciprofloxacin. However, they were resistant to amoxycilline + clavulanic acid, penicillin, erythromycine, oxytetracycline and cefuroxime sodium. PMID- 14535933 TI - Bacteriological status of canine milk and septicaemia in neonatal puppies--a retrospective study. AB - Results of the bacteriological examination of milk samples from 46 bitches were evaluated retrospectively, and correlated with findings of the bacteriological examination of organs from dead, septicaemic puppies in their litters (n = 33). The aim of this study was to investigate, in how many cases of clinical and subclinical mastitis of the bitches, the same bacteria can be detected in their septicaemic neonates. One group of lactating bitches was clinically healthy (group I, n = 38), whereas in eight bitches different puerperal disorders were found (group II). Twenty-five septicaemic puppies were from group I, eight from group II. Out of a broad spectrum of bacteria isolated from the milk of clinically healthy and diseased bitches, only Escherichia coli, Klebsiella pneumoniae and/or ss-haemolytic (haem.) Streptococcus sp. could be isolated from organs of their septicaemic puppies. This was the case in three bitches with mastitis and in one clinically healthy bitch only. Staphylococcus intermedius, although frequently isolated from canine milk, does not seem to be a cause of septicaemia in neonates. It is assumed that in most cases of neonate septicaemia, bacteria from the bitches' milk are not the primary cause. PMID- 14535934 TI - Isolation of Mycoplasmas from nasal swabs of calves affected with respiratory diseases and antimicrobial susceptibility of their isolates. AB - Nasal swabs of 293 calves were examined for Mycoplasma. The samples were collected from calves affected with respiratory diseases on 71 farms in various parts of Japan between 1996 and 1997. Mycoplasma bovirhinis was isolated from 47 of 293 calves (16.0%). Mycoplasma alkalescens, M. bovis, M. arginini, M. bovigenitalium and Acholeplasma spp. were isolated from 19 (6.5%), seven (2.4%), four (1.4%), four (1.4%) and 18 (6.1%) calves, respectively. Pasteurella multocida and P. haemolytica were isolated from 60% of Mycoplasma-positive calves. However, other bacteria were not isolated from calves. To evaluate the antimicrobial susceptibility of their isolates, 68 M. bovirhinis, 21 M. alkalescens and 10 M. bovis strains were examined for 12 antimicrobial agents. All isolates showed higher susceptibility to tiamulin than to the other drugs used in the study. However, erythromycin had no effect on any of the Mycoplasma strains studied. The field isolates were less susceptible than the type strains to some drugs, such as spiramycin, oxytetracycline and tylosin. PMID- 14535935 TI - The effect of commercial steam pasteurization on the levels of Enterobacteriaceae and Escherichia coli on naturally contaminated beef carcasses. AB - The aim of the study was to assess the reduction achieved by steam pasteurization of beef carcasses of Escherichia coli, Enterobacteriaceae and total aerobic mesophilic plate counts (APCs). In total, 30 carcass halves were exposed to steam pasteurization (90 degrees C, 10 s exposure time) and the 30 corresponding carcass halves remained as untreated controls. The neck, midline and rump were sampled on each carcass half. Significant reductions in E. coli incidence (P < 0.05) and counts, 0.5 log10 CFU 1000 cm(-2) (P < 0.05), were observed on rump sites only. Significant reductions (>0.8 log10 CFU 1000 cm(-2)) of Enterobacteriaceae were observed at all carcass sites sampled (P < 0.05). Enterobacteriaceae reductions (>2 log10 CFU 1000 cm(-2)) were highly significant at the more contaminated sites (P < 0.001). Reductions in total APCs were inconsistent. Steam pasteurization significantly reduced the level of E. coli and Enterobacteriaceae at more contaminated sites, but did not result in complete decontamination. Therefore, steam pasteurization should be classed as an aid to hygienic beef processing, but not as a critical control point. PMID- 14535936 TI - Role of birds in transmission of classical swine fever virus. AB - Active transmission of classical swine fever virus (CSFV) was studied in six birds (five ravens, one hooded crow) and two laying hens. Cloacal swabs, blood and organs of birds and hens as well as blood and organ samples of pigs which had been fed with faeces derived from CSFV infected birds or which had come in contact with faeces of infected hens were negative for CSFV. None of the animals seroconverted during the study. This result demonstrates that active virus transmission by these animals is unlikely. Dissemination of CSFV from wild boar to domestic pigs is discussed. PMID- 14535937 TI - Genotyping of Clostridium perfringens isolated from domestic and exotic ruminants and swine. AB - Clostridium perfringens types A, B, C, D and E are known to cause severe enteritis/enterotoxaemia and diseases (especially caused by type A) belonging to the gas oedema complex in many species. Samples from the small intestine as well as faeces of domestic and exotic animals suffering from enterotoxaemic signs or having died within days after first occurance of toxaemia were submitted for typing C. perfringens toxovars by multiplex PCR. The following species have been investigated: domestic sheep (Ovis ammon; n = 10), domestic goat (Capra aegagrus hircus; n = 26), Japanese serow (Capricornis sumatraensis; n = 4), lechwe waterbuck (Hydrotragus leche; n = 1), blackbuck (Antilope cervicapra; n = 1), European reindeer (Rangifer tarandus tarandus; n = 4), domestic swine (Sus scrofa; n = 52), and collared peccary (Tayassu albirostris; n = 1). Interestingly, the predominant C. perfringens toxovar in domestic sheep was type A. This toxovar could also be diagnosed in all reindeer, in three Japanese serows, one lechwe waterbuck and most pigs (n = 47), the majority of those being at suckling age. Type D was the most prevalent toxovar (n = 18) in domestic goats, but also types A and E could be identified as pathogens in this species. Type C could only be found in domestic swine (n = 5) and in one case of clostridiosis in a Japanese serow. Two cases of enterotoxaemia in goats, one case in reindeer, and a single case in blackbuck and collared peccary were caused by C. perfringens type E. Genotyping of C. perfringens is recommended before starting vaccination programmes as it could be shown, that the importance of specific toxovars has been underestimated in specific species and/or age groups. PMID- 14535938 TI - Membrane receptors for oestrogen in the brain. AB - Oestrogen is important for the development of neuroendocrine centres and other neural networks including limbic and motor systems. Later in adulthood, oestrogen regulates the functional performance of different neural systems and is presumably implicated in the modulation of cognitive efficiency. Although still a matter of controversial discussion, clinical and experimental studies point at a potential neuroprotective role of oestrogen. Concerning the concept of cellular oestrogen action, it is undisputed that it comprises the binding and activation of nuclear receptors. The last decades have, however, immensely broadened the spectrum of steroid signalling within a cell. Novel steroid-activated intracellular signalling mechanisms were described which are usually termed 'non classical' or 'non-genomic'. The brain appears to be a rich source of this new mode of oestrogen action. Studies from the past years have pinpointed non classical oestrogen effects in many CNS regions. All available data support the view that non-classical oestrogen action requires interactions with putative membrane binding sites/receptors. In this article, we aim at compiling the most recent findings on the nature and identity of membrane oestrogen receptors with respect to the brain. We also attempt to turn readers attention to the coupling of these 'novel' receptors to distinct intracellular signalling pathways. PMID- 14535939 TI - In through the out door: nuclear localization of the regulators of G protein signaling. AB - The regulators of G protein signaling (RGS) are an extraordinary class of diverse multifunctional signaling proteins best known for their potent capacity to down regulate the activity of Galpha subunits at the plasma membrane. In certain circumstances, some RGS proteins undergo translocation to the nucleus or plasma membrane from the cytoplasm. Translocation demonstrates a potentially dynamic alternative mechanism for Galpha subunit or effector regulation. The nuclear localization of the regulators of G protein signaling proteins further suggests these proteins possess even greater functional heterogeneity than that envisioned previously, as regulators of transcription and cell cycle control. PMID- 14535940 TI - Coupling gene chip analyses and rat genetic variances in identifying potential target genes that may contribute to neuropathic allodynia development. AB - Genetic factors and nerve injury-induced changes of gene expression in sensory neurons are potential contributors to tactile allodynia, a neuropathic pain state manifested as hypersensitivity to innocuous mechanical stimulation. To uncover genes relevant to neuropathic allodynia, we analyzed gene expression profiles in dorsal root ganglia (DRG) of spinal nerve-ligated Harlan and Holtzman Sprague Dawley rats, strains with different susceptibilities to neuropathic allodynia. Using Affymetrix gene chips, we identified genes showing differential basal-level expression in these strains without injury-induced regulation. Of more than 8000 genes analyzed, less than 180 genes in each strain were regulated after injury, and 19-22% of that was regulated in a strain-specific manner. Importantly, we identified functionally related genes that were co-regulated post injury in one or both strains. In situ hybridization and real-time PCR analyses of a subset of identified genes confirmed the patterns of the microarray data, and the former also demonstrated that injury-induced changes occurred, not only in neurons, but also in non-neuronal cells. Together, our studies provide a global view of injury plasticity in DRG of these rat stains and support a plasticity-based mechanism mediating variations in allodynia susceptibility, thus providing a source for further characterization of neuropathic pain-relevant genes and potential pathways. PMID- 14535941 TI - Marked disparity between age-related changes in dopamine and other presynaptic dopaminergic markers in human striatum. AB - Because age-related changes in brain dopaminergic innervation are assumed to influence human disorders involving dopamine (DA), we measured the levels of several presynpatic DAergic markers [DA, homovanillic acid, tyrosine hydroxylase (TH), aromatic L-amino acid decarboxylase (AADC), vesicular monoamine transporter 2 (VMAT2), and dopamine transporter (DAT)] in post-mortem human striatum (caudate and putamen) from 56 neurologically normal subjects aged 1 day to 103 years. Striatal DA levels exhibited pronounced (2- to 3-fold) post-natal increases through adolescence and then decreases during aging. Similarly, TH and AADC increased almost 100% during the first 2 post-natal years; however, the levels of TH and, to a lesser extent, AADC then declined to adult levels by approximately 30 years of age. Although VMAT2 and DAT levels closely paralleled those of TH, resulting in relatively constant TH to transporter ratios during development and aging, a modest but significant decline (13%) in DAT levels was observed in only caudate during aging. This biphasic post-natal pattern of the presynaptic markers suggests that striatal DAergic innervation/neuropil appears to continue to develop well past birth but appears to become overelaborated and undergo regressive remodeling during adolescence. However, during adulthood, a striking discrepancy was observed between the loss of DA and the relative preservation of proteins involved in its biosynthesis and compartmentation. This suggests that declines in DA-related function during adulthood and senescence may be explained by losses in DA per se as opposed to DAergic neuropil. PMID- 14535942 TI - Stroke-induced progenitor cell proliferation in adult spontaneously hypertensive rat brain: effect of exogenous IGF-1 and GDNF. AB - Progenitor cells in the dentate gyrus of hippocampus (DG) and the subventricular zone of lateral ventricles (SVZ) generate new neurons throughout the life of mammals. Cerebral ischemia increases this basal progenitor cell proliferation. The present study evaluated the time frame of proliferation, length of survival and the phenotypes of the new cells formed after transient middle cerebral artery occlusion (MCAO) in adult spontaneously hypertensive rats. Compared to sham controls, ischemic rats showed a significantly higher number of newly proliferated cells (as defined by BrdU immunostaining) in both the DG (by fourfold, p < 0.05) and the SVZ (by twofold, p < 0.05). DG showed increased proliferation only in the first week of reperfusion and 49% of the cells formed in this period survived to the end of third week. Whereas, SVZ showed a continuous proliferation up to 3 weeks after MCAO, but the cells formed survived for less than a week. In both DG and SVZ, at the end of the first week of reperfusion, majority of the BrdU-positive (BrdU+) cells were immature neurons (DCX positive). In the DG, 28% of the cells formed in the first week after MCAO mature into neurons (NeuN positive). The ischemic cortex and striatum showed several BrdU+ cells which were ED-1 positive microglia/macrophages. At 1 week of reperfusion, MCAO-induced progenitor cell proliferation in the ipsilateral DG was significantly increased by i.c.v. infusion of IGF-1 (by 127 +/- 14%, p < 0.05) and GDNF (by 91 +/- 5%, p < 0.05), compared to vehicle. In the growth factor treated rats subjected to transient MCAO, several BrdU+ cells formed in the first week survived up to the third week. PMID- 14535943 TI - Induction by beta-bungarotoxin of apoptosis in cultured hippocampal neurons is mediated by Ca(2+)-dependent formation of reactive oxygen species. AB - The component of the venom of the Taiwanese banded krait Bungarus multicinctus, beta-bungarotoxin (beta-BuTx), acts as an extremely potent inducer of neuronal apoptosis when applied to rat hippocampal cultures. While induction of cell death is dependent on toxin binding to voltage-activated K+ channels and subsequent internalization, the pro-apoptotic signals triggered by picomolar concentrations of beta-BuTx are not understood. Following toxin binding, a dramatic increase in intracellular Ca2+ became detectable after 30 min, and in reactive oxygen species (ROS) after 3-4 h. Conversely, Ca2+ chelators, radical quenchers and antioxidants efficiently antagonized beta-BuTx induced apoptosis. As shown for the antioxidant 2,3-dihydroxybenzoic acid, analysis by matrix assisted laser desorbtion-time of flight (MALDI-TOF) mass spectrometry excluded the protective effects to be due to reductive cleavage of the toxic beta-BuTx dimer. Inhibitors of the intracellular antioxidant defence system enhanced neuronal susceptibility to beta-BuTx, supporting the essential role of ROS in beta-BuTx-initiated apoptosis. Cell damage was accompanied by an accumulation of markers of oxidative cell stress, phospholipid hydroxyperoxides and the lipid peroxidation product, malonyl dialdehyde. These observations indicate that beta-BuTx-induced cell death resulted from an intracellular signalling cascade involving subsequent stages of a dramatic rise in free Ca2+, the accumulation of ROS, membrane lipid peroxidation and, finally, apoptosis. PMID- 14535944 TI - Amyloid beta-protein (Abeta)1-40 protects neurons from damage induced by Abeta1 42 in culture and in rat brain. AB - Previously, we found that amyloid beta-protein (Abeta)1-42 exhibits neurotoxicity, while Abeta1-40 serves as an antioxidant molecule by quenching metal ions and inhibiting metal-mediated oxygen radical generation. Here, we show another neuroprotective action of nonamyloidogenic Abeta1-40 against Abeta1-42 induced neurotoxicity in culture and in vivo. Neuronal death was induced by Abeta1-42 at concentrations higher than 2 microm, which was prevented by concurrent treatment with Abeta1-40 in a dose-dependent manner. However, metal chelators did not prevent Abeta1-42-induced neuronal death. Circular dichroism spectroscopy showed that Abeta1-40 inhibited the beta-sheet transformation of Abeta1-42. Thioflavin-T assay and electron microscopy analysis revealed that Abeta1-40 inhibited the fibril formation of Abeta1-42. In contrast, Abeta1-16, Abeta25-35, and Abeta40-1 did not inhibit the fibril formation of Abeta1-42 nor prevent Abeta1-42-induced neuronal death. Abeta1-42 injection into the rat entorhinal cortex (EC) caused the hyperphosphorylation of tau on both sides of EC and hippocampus and increased the number of glial fibrillary acidic protein (GFAP)-positive astrocytes in the ipsilateral EC, which were prevented by the concurrent injection of Abeta1-40. These results indicate that Abeta1-40 protects neurons from Abeta1-42-induced neuronal damage in vitro and in vivo, not by sequestrating metals, but by inhibiting the beta-sheet transformation and fibril formation of Abeta1-42. Our data suggest a mechanism by which elevated Abeta1 42/Abeta1-40 ratio accelerates the development of Alzheimer's disease (AD) in familial AD. PMID- 14535945 TI - Fe(II)-induced DNA damage in alpha-synuclein-transfected human dopaminergic BE(2) M17 neuroblastoma cells: detection by the Comet assay. AB - Lewy bodies in the brains of patients with Parkinson's disease (PD) contain aggregates of alpha-synuclein (alpha-syn). Missense mutations (A53T or A30P) in the gene encoding alpha-syn are responsible for rare, inherited forms of PD. In this study, we explored the susceptibility of untransfected human dopaminergic BE(2)-M17 neuroblastoma cells, cells transfected with vector only, or cells transfected with wild-type alpha-syn, A30P alpha-syn or A53T alpha-syn to Fe(II) induced DNA damage in the form of single-strand breaks (SSBs). DNA SSBs were detected following 2-h treatments with various concentrations of Fe(II) (0.01 100.0 microm), using the alkaline single cell-gel electrophoresis ('Comet') assay and quantified by measuring comet tail length (CTL) microm). Fe(II) treatment induced significant increases in CTL in cells transfected with A30P alpha-syn or A53T alpha-syn, even at the lowest concentrations of Fe(II) tested. In comparison, untransfected cells, vector control cells or cells transfected with wild-type alpha-syn exhibited increases in SSBs only when exposed to concentrations of 1.0 microm Fe(II) and above. Even when exposed to higher concentrations (10.0-100.0 microm) of Fe(II), untransfected cells, vector control cells or cells transfected with wild-type alpha-syn were less susceptible to DNA damage induction than cells transfected with A30P alpha-syn or A53T alpha-syn. Incorporation of DNA-repair inhibitors, hydroxyurea and cytosine arabinoside, enhanced the sensitivity of DNA damage detection. Susceptibility to Fe(II) induced DNA damage appeared to be dependent on alpha-syn status because cells transfected with wild-type alpha-syn or A53T alpha-syn were equally susceptible to the damaging effects of the mitochondrial respiratory chain inhibitor rotenone. Overall, our data are suggestive of an enhanced susceptibility to the toxic effects of Fe(II) in neuroblastoma cells transfected with mutant alpha-syn associated with inherited forms of PD. PMID- 14535946 TI - Potent activation of dopamine D3/D2 heterodimers by the antiparkinsonian agents, S32504, pramipexole and ropinirole. AB - Recombinant, human dopamine D3 and D2 receptors form functional heterodimers upon co-expression in COS-7 cells. Herein, actions of the antiparkinsonian agents, S32504, ropinirole and pramipexole, at D3/D2L heterodimers were compared to their effects at the respective monomers and at split, chimeric D3trunk/D2tail and D2trunk/D3tail receptors: the trunk incorporated transmembrane domains (TDs) I-V and the tail TDs VI and VII. In binding assays with the antagonist [3H]nemonapride, all agonists were potent ligands of D3 receptors showing, respectively, 100-, 18- and 56-fold lower affinity at D2L receptors, mimicking the selective D3 receptor antagonist, S33084 (100-fold). At D3trunk/D2tail receptors, except for ropinirole, all drugs showed lower affinities than at D3 sites, whereas for D2trunk/D3tail receptors, affinities of all drugs were higher than at D2L sites. The proportion of high affinity binding sites recognized by S32504, pramipexole and ropinirole in membranes derived from cells co-expressing D3 and D2L sites was higher than in an equivalent mixture of membranes from cells expressing D3 or D2L sites, consistent with the promotion of heterodimer formation. In contrast, the percentage of high and low affinity sites (biphasic isotherms) recognized by S33084 was identical. Functional actions were determined by co-transfection of a chimeric adenylyl cyclase (AC)-V/VI insensitive to D3 receptors. Accordingly, D3 receptor-transfected cells were irresponsive whereas, in D2L receptor-transfected cells, agonists suppressed forskolin-stimulated cAMP production with modest potencies. In cells co-transfected with D3 and D2L receptors, S32504, ropinirole and pramipexole potently suppressed AC-V/VI with EC50s 33-, 19- and 11-fold lower than at D2L receptors, respectively. S32504 also suppressed AC-V/VI activity at split D3trunk/D2tail and D2trunk/D3tail chimeras transfected into COS-7 cells. In conclusion, antiparkinson agents behave as potent agonists at D3/D2'heterodimers', though any role in their actions in vivo remains to be demonstrated. PMID- 14535947 TI - Mass spectrometric investigation of the neuropeptide complement and release in the pericardial organs of the crab, Cancer borealis. AB - The crustacean stomatogastric ganglion (STG) is modulated by both locally released neuroactive compounds and circulating hormones. This study presents mass spectrometric characterization of the complement of peptide hormones present in one of the major neurosecretory structures, the pericardial organs (POs), and the detection of neurohormones released from the POs. Direct peptide profiling of Cancer borealis PO tissues using matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) revealed many previously identified peptides, including proctolin, red pigment concentrating hormone (RPCH), crustacean cardioactive peptide (CCAP), several orcokinins, and SDRNFLRFamide. This technique also detected corazonin, a well-known insect hormone, in the POs for the first time. However, most mass spectral peaks did not correspond to previously known peptides. To characterize and identify these novel peptides, we performed MALDI postsource decay (PSD) and electrospray ionization (ESI) MS/MS de novo sequencing of peptides fractionated from PO extracts. We characterized a truncated form of previously identified TNRNFLRFamide, NRNFLRFamide. In addition, we sequenced five other novel peptides sharing a common C-terminus of RYamide from the PO tissue extracts. High K+ depolarization of isolated POs released many peptides present in this tissue, including several of the novel peptides sequenced in the current study. PMID- 14535948 TI - Co-ordinated expression of 5-HT2C receptors with the NCX1 Na+/Ca2+ exchanger in histaminergic neurones. AB - The different roles of Na+/Ca2+ (NCX) exchangers and Na+/Ca2+/K+ (NCKX) exchangers in regulation of the ionic homeostasis in neurones are poorly understood. We have previously shown that serotonin excites histaminergic tuberomamillary (TM) neurones by activation of 5-HT2C-receptors and Na+/Ca2+ exchange. With the help of single-cell RT-PCR (sc-RT-PCR) we have now determined the coexpression pattern of different subtypes of NCX and NCKX with serotonin receptors. The majority of TM neurones express NCX1, NCX2 and NCKX3. Serotonin 2C receptor-mRNA was detected in 70% while 5-HT2A mRNA was found in only 10% of TM neurones. In all neurones expressing the 5-HT2C receptor NCX1-mRNA was present. Double immunostaining revealed the presence of the NCX1 protein in histidine decarboxylase-positive neurones. In the majority of TM neurones one or two out of five isoforms, NCX1.4, NCX1.5, NCX1.7, NCX1.14, NCX1.15, were detected by cDNA sequencing and/or by restriction analysis. The alternative splicing region is important for the Ca2+ sensitivity and presumably for the modulation of NCX1 function by second messengers. We conclude that several exchanger-subtypes can be coexpressed in single neurones and that TM cells are heterogeneous with respect to their calcium homeostasis regulation. PMID- 14535949 TI - A novel ubiquitin-specific protease, synUSP, is localized at the post-synaptic density and post-synaptic lipid raft. AB - Recent reports suggest an important role for protein ubiquitination in synaptic plasticity. We cloned, from the rat brain, a novel gene that encoded an ubiquitin specific protease (USP), and termed this protein synaptic ubiquitin-specific protease (synUSP, GenBankTM Accession no. AB073880). The homologous human gene was mapped to a locus on chromosome 1p36.12. The deduced synUSP protein consisted of 1036 amino acids, and possessed an ubiquitin-like domain at the C-terminus, Cys- and His-boxes, leucine zipper motifs, and six amino acid-repeats of L/ILCPHG. The protein possessed de-ubiquitinating activity toward a model substrate, as expected from its sequence. The protein of 125 kDa was present in the rat brain; in particular, it was enriched in the post-synaptic density and the dendritic lipid raft fractions. The immunostaining of cortical neurons confirmed the post-synaptic localization. The mRNA for synUSP was localized to dendrites, as well as somas, of neuronal cells. Thus, both the mRNA and the protein were localized in the post-synaptic compartments. These results suggest a regulatory mechanism for the ubiquitin-related system at the post-synaptic sites. PMID- 14535950 TI - Polarized actin bundles formed by human fascin-1: their sliding and disassembly on myosin II and myosin V in vitro. AB - Fascin-1 is a putative bundling factor of actin filaments in the filopodia of neuronal growth cones. Here, we examined the structure of the actin bundle formed by human fascin-1 (actin/fascin bundle), and its mode of interaction with myosin in vitro. The distance between cross-linked filaments in the actin/bundle was 8-9 nm, and the bundle showed the transverse periodicity of 36 nm perpendicular to the bundle axis, which was confirmed by electron microscopy. Decoration of the actin/fascin bundle with heavy meromyosin revealed that the arrowheads of filaments in the bundle pointed in the same direction, indicating that the bundle has polarity. This result suggested that fascin-1 plays an essential role in polarity of actin bundles in filopodia. In the in vitro motility assay, actin/fascin bundles slid as fast as single actin filaments on myosin II and myosin V. When myosin was attached to the surface at high density, the actin/fascin bundle disassembled to single filaments at the pointed end of the bundle during sliding. These results suggest that myosins may drive filopodial actin bundles backward by interacting with actin filaments on the surface, and may induce disassembly of the bundle at the basal region of filopodia. PMID- 14535951 TI - Haloperidol impairs auditory filial imprinting and modulates monoaminergic neurotransmission in an imprinting-relevant forebrain area of the domestic chick. AB - In vivo microdialysis and behavioural studies in the domestic chick have shown that glutamatergic as well as monoaminergic neurotransmission in the medio rostral neostriatum/hyperstriatum ventrale (MNH) is altered after auditory filial imprinting. In the present study, using pharmaco-behavioural and in vivo microdialysis approaches, the role of dopaminergic neurotransmission in this juvenile learning event was further evaluated. The results revealed that: (i) the systemic application of the potent dopamine receptor antagonist haloperidol (7.5 mg/kg) strongly impairs auditory filial imprinting; (ii) systemic haloperidol induces a tetrodotoxin-sensitive increase of extracellular levels of the dopamine metabolite, homovanillic acid, in the MNH, whereas the levels of glutamate, taurine and the serotonin metabolite, 5-hydroxyindole-3-acetic acid, remain unchanged; (iii) haloperidol (0.01, 0.1, 1 mm) infused locally into the MNH increases glutamate, taurine and 5- hydroxyindole-3-acetic acid levels in a dose dependent manner, whereas homovanillic acid levels remain unchanged; (iv) systemic haloperidol infusion reinforces the N-methyl-d-aspartate receptor mediated inhibitory modulation of the dopaminergic neurotransmission within the MNH. These results indicate that the modulation of dopaminergic function and its interaction with other neurotransmitter systems in a higher associative forebrain region of the juvenile avian brain displays similar neurochemical characteristics as the adult mammalian prefrontal cortex. Furthermore, we were able to show that the pharmacological manipulation of monoaminergic regulatory mechanisms interferes with learning and memory formation, events which in a similar fashion might occur in young or adult mammals. PMID- 14535952 TI - The first and third intracellular loops together with the carboxy terminal tail of the delta-opioid receptor contribute toward functional interaction with Galpha16. AB - Opioid peptides exert their regulatory effects on both central and peripheral nervous systems via multiple opioid receptors that are linked to seemingly identical sets of guanine nucleotide-binding regulatory proteins (G proteins). In contrast to the mu-opioid receptor, the delta-opioid receptor can efficiently stimulate phospholipase C via G16. We used a series of mu/delta-opioid receptor chimeras to examine the involvement of intracellular receptor domains in the recognition of G16. After ascertaining that the chimeras can bind opioid ligands with high affinity and elicit inhibition of adenylyl cyclase, COS-7 cells were cotransfected with cDNAs encoding Galpha16 and a mu/delta-opioid receptor chimera and assayed for [D-Ala2,D-Leu5]enkephalin-induced stimulation of phospholipase C. Our results indicate that (i) the carboxy terminal tail of the delta-opioid receptor is necessary but insufficient for conferring coupling to Galpha16, (ii) the third inner loop together with the carboxy terminal tail of the delta-opioid receptor can provide sufficient contact domains for Galpha16, and (iii) the first inner loop of the delta-opioid receptor, in particular Leu80, as well as the fifth transmembrane domain and/or the third extracellular loop may also contribute in defining the fidelity of interaction between the delta-opioid receptor and Galpha16. These results indicate that efficient coupling of the delta-opioid receptor to Galpha16 requires the participation of most of the intracellular regions, including the first intracellular loop. PMID- 14535953 TI - Expression of BDNF mRNA in substantia nigra is dependent on target integrity and independent of neuronal activation. AB - We have analyzed the regulation of brain-derived neurotrophic factor (BDNF) mRNA expression in the nigrostriatal system following neurotoxin ablation of striatal targets by means of kainate (KA) or quinolinic acid (QA) injections. Loss of nigral target cells in the striatum was accompanied by significant induction of BDNF mRNA levels in the ipsilateral substantia nigra (SN) at 12 and 24 h post lesion. Dual tyrosine hydroxylase (TH) and BDNF mRNA in situ hybridization (ISH) confirmed the dopaminergic nature of the BDNF mRNA expressing cells. Analysis of neuronal activity in terms of cFos mRNA expression demonstrated intense induction of this marker in the ipsilateral SN pars reticulata (SNPR), but not in SN pars compacta. Dual glutamic acid decarboxylase (GAD) and cFos mRNA ISH confirmed this view. Colchicine injections into the medial forebrain bundle to specifically disrupt neuronal trafficking between SN and striatum induced BDNF mRNA levels in the ipsilateral SNPC, thus demonstrating that nigral expression of BDNF mRNA is dependent of striatal target tissue. In addition, we found significant elevations of BDNF in the subthalamic nucleus following striatal excitotoxic lesion, which may bring novel roles of BDNF in the basal ganglia complex. PMID- 14535955 TI - Caspase cleavage of the amyloid precursor protein modulates amyloid beta-protein toxicity. AB - The amyloid beta-protein precursor (APP) is proteolytically cleaved to generate the amyloid beta-protein (Abeta), the principal constituent of senile plaques found in Alzheimer's disease (AD). In addition, Abeta in its oligomeric and fibrillar forms have been hypothesized to induce neuronal toxicity. We and others have previously shown that APP can be cleaved by caspases at the C-terminus to generate a potentially cytotoxic peptide termed C31. Furthermore, this cleavage event and caspase activation were increased in the brains of AD, but not control, cases. In this study, we show that in cultured cells, Abeta induces caspase cleavage of APP in the C-terminus and that the subsequent generation of C31 contributes to the apoptotic cell death associated with Abeta. Interestingly, both Abeta toxicity and C31 pathway are dependent on the presence of APP. Both APP-dependent Abeta toxicity and C31-induced apoptotic cell death involve apical or initiator caspases-8 and -9. Our results suggest that Abeta-mediated toxicity initiates a cascade of events that includes caspase activation and APP cleavage. These findings link C31 generation and its potential cell death activity to Abeta cytotoxicity, the leading mechanism proposed for neuronal death in AD. PMID- 14535954 TI - Selective gamma-hydroxybutyric acid receptor ligands increase extracellular glutamate in the hippocampus, but fail to activate G protein and to produce the sedative/hypnotic effect of gamma-hydroxybutyric acid. AB - Two gamma-hydroxybutyric acid (GHB) analogues, trans-gamma-hydroxycrotonic acid (t-HCA) and gamma-(p-methoxybenzyl)-gamma-hydroxybutyric acid (NCS-435) displaced [3H]GHB from GHB receptors with the same affinity as GHB but, unlike GHB, failed to displace [3H]baclofen from GABAB receptors. The effect of the GHB analogues, GHB and baclofen, on G protein activity and hippocampal extracellular glutamate levels was compared. While GHB and baclofen stimulated 5'-O-(3 [35S]thiotriphospate) [35S]GTPgammaS binding both in cortex homogenate and cortical slices, t-HCA and NCS-435 were ineffective up to 1 mm concentration. GHB and baclofen effect was suppressed by the GABAB antagonist CGP 35348 but not by the GHB receptor antagonist NCS-382. Perfused into rat hippocampus, 500 nm and 1 mm GHB increased and decreased extracellular glutamate levels, respectively. GHB stimulation was suppressed by NCS-382, while GHB inhibition by CGP 35348. t-HCA and NCS-435 (0.1-1000 microm) locally perfused into hippocampus increased extracellular glutamate; this effect was inhibited by NCS-382 (10 microm) but not by CGP 35348 (500 microm). The results indicate that GHB-induced G protein activation and reduction of glutamate levels are GABAB-mediated effects, while the increase of glutamate levels is a GHB-mediated effect. Neither t-HCA nor NCS 435 reproduced GHB sedative/hypnotic effect in mice, confirming that this effect is GABAB-mediated. The GHB analogues constitute important tools for understanding the physiological role of endogenous GHB and its receptor. PMID- 14535956 TI - 15-deoxy-delta12,14-prostaglandin J2 regulates the functional state and the survival of microglial cells through multiple molecular mechanisms. AB - We have previously reported that rat primary microglial cultures express the nuclear receptor peroxisome proliferator-activated receptor-gamma (PPAR-gamma) and that several functions associated with the activation of these cells, including nitric oxide (NO) and tumor necrosis factor-alpha synthesis, are down regulated by 15-deoxy-delta12,14-prostaglandin J2 (15d-PGJ2) and ciglitazone, two specific PPAR-gamma agonists. Here we demonstrate that microglial cells not only express a functionally active PPAR-gamma, but also synthesize large amounts of 15d-PGJ2 upon stimulation with lipopolysaccharide (LPS). In addition, we show that, although 15d-PGJ2 and ciglitazone were equally effective in reducing microglial activation when used at 1-5 microm concentrations, 15d-PGJ2, but not of ciglitazone, reduced PGE2 production at low concentration (0.1 microm) and induced a time-dependent microglial impairment and apoptosis at high concentration (10 microm). Interestingly, the inhibition of PGE2 production was achieved mainly through the inhibition of cycloxygenase-2 enzymatic activity, as the expression of this enzyme and that of the microsomal isoform of PGE synthase remained unaltered. These findings suggest that 15d-PGJ2 affects the functional state and the survival of activated microglia through mechanisms only in part dependent on PPAR-gamma and that the concentration of 15d-PGJ2 is crucial in determining the particular microglial function affected. PMID- 14535957 TI - Antisense peptide nucleic acid-mediated knockdown of the p75 neurotrophin receptor delays motor neuron disease in mutant SOD1 transgenic mice. AB - Re-expression of the death-signalling p75 neurotrophin receptor (p75NTR) is associated with injury and neurodegeneration in the adult nervous system. The induction of p75NTR expression in mature degenerating spinal motor neurons of humans and transgenic mice with amyotrophic lateral sclerosis (ALS) suggests a role of p75NTR in the progression of motor neuron disease (MND). In this study, we designed, synthesized and evaluated novel antisense peptide nucleic acid (PNA) constructs targeting p75NTR as a potential gene knockdown therapeutic strategy for ALS. An 11-mer antisense PNA directed at the initiation codon, but not downstream gene sequences, dose-dependently inhibited p75NTR expression and death signalling by nerve growth factor (NGF) in Schwann cell cultures. Antisense phosphorothioate oligonucleotide (PS-ODN) sequences used for comparison failed to confer such inhibitory activity. Systemic intraperitoneal administration of this antisense PNA to mutant superoxide dismutase 1 (SOD1G93A) transgenic mice significantly delayed locomotor impairment and mortality compared with mice injected with nonsense or scrambled PNA sequences. Reductions in p75NTR expression and subsequent caspase-3 activation in spinal cords were consistent with increased survival in antisense PNA-treated mice. The uptake of fluorescent labelled antisense PNA in the nervous system of transgenic mice was also confirmed. This study suggests that p75NTR may be a promising antisense target in the treatment of ALS. PMID- 14535958 TI - Primary cortical neuronal cultures reduce cellular energy utilization during anoxic energy deprivation. AB - It has been widely hypothesized that neurons reduce cellular energy use in response to periods of energy deprivation. To test this hypothesis, we measured rates of energy use under normoxia and anoxia in immature (6 days in vitro) and mature (13 days in vitro) neuronal cultures. During anoxic incubation immature and mature cultures reduced cellular energy use by 80% and 45%, respectively. Reduced cellular energy use dramatically affected ATP depletion in neuronal cultures under anoxia. Intracellular ATP stores were expected to deplete within 3 min of anoxia. However, ATP was maintained at decreased but stabilized concentrations for at least 3 h. The capacity of neuronal cultures to reduce cellular energy use during anoxia correlated with their sensitivity towards simulated ischemia. Immature cultures, with the largest capacity to reduce cellular energy use, survived simulated ischemia 2.5 times longer than mature cultures. The addition of glutamate receptor antagonists to mature cultures further decreased cellular energy use during anoxia and significantly extended their survival time under simulated ischemia. This study verifies that primary cortical neuronal cultures reduce cellular energy use during energy deprivation. Additionally, we show that maturation of glutamate receptor activity increases non-depressible energy demand in neuronal cultures. PMID- 14535959 TI - Acetylcholinesterase level and molecular isoforms are altered in brain of Reelin Orleans mutant mice. AB - In this study we examined changes in acetylcholinesterase (AChE) pattern in the brain of adult Reelin Orleans (RelnOrl) homozygous mutant mice. The AChE histochemistry firstly revealed an abnormal distribution of AChE-positive cells in several areas of the reeler brain, including cortices; the strongest labelling was observed in cerebellum and hippocampus when compared with controls. Biochemical determinations demonstrated an increase of 80-90% in AChE specific activity from cerebellar and hippocampal extracts. We also report that the AChE tetrameric form (G4) was selectively increased in the RelnOrl brain. The relationship between AChE and Reelin and suggested morphogenetic functions are also discussed. PMID- 14535960 TI - Identification of Tau and MAP2 as novel substrates of Rho-kinase and myosin phosphatase. AB - Rho-kinase and myosin phosphatase are implicated in the phosphorylation-state of myosin light chain downstream of Rho, which is thought to induce smooth muscle contraction and stress fibre formation in non-muscle cells. Here, we found that microtubule-associated proteins, Tau and MAP2, interacted with the myosin-binding subunit (MBS) of myosin phosphatase, and were the possible substrates of both Rho kinase and myosin phosphatase. We determined the phosphorylation sites of Tau (Thr245, Thr377, Ser409) and MAP2 (Ser1796) by Rho-kinase. We also found that Rho kinase phosphorylated Tau at Ser262 to some extent. Phosphorylation by Rho-kinase decreased the activity of Tau to promote microtubule assembly in vitro. Substitutions of Ala for Ser/Thr at the phosphorylation sites of Tau (Tau-AAA) did not affect the activity to promote microtubule assembly, while substitutions of Asp for Ser/Thr (Tau-DDD), which are expected to mimic the phosphorylation state of Tau, slightly reduced the activity. When Tau, or mutated forms of Tau, were expressed in PC12 cells, followed by treatment with cytochalasin D, they promoted extension of the cell process in a cytochalasin-dependent manner. However, Tau-DDD showed the weaker activity in this capacity than wild-type Tau or Tau-AAA. These results suggest that the phosphorylation-state of these residues of Tau affects its activity both in vitro and in vivo. Thus, it is likely that the Rho-kinase/MBS pathway regulates not only the actin-myosin system but also microtubule dynamics. PMID- 14535961 TI - GABAC receptor agonist suppressed ammonia-induced apoptosis in cultured rat hippocampal neurons by restoring phosphorylated BAD level. AB - Ammonia-induced apoptosis and its prevention by GABAC receptor stimulation were examined using primary cultured rat hippocampal neurons. Ammonia (0.5-5 mm NH4Cl) dose-dependently induced apoptosis in pyramidal cell-like neurons as assayed by double staining with Hoechst 33258 and anti-neurofilament antibody. A GABAC receptor agonist, cis-4-aminocrotonic acid (CACA, 200 microm), but not GABAA and GABAB receptor agonists, muscimol (10 micro m) and baclofen (50 microm), respectively, inhibited the ammonia (2 mm)-induced apoptosis, and this inhibition was abolished by a GABAC receptor antagonist (1,2,5,6-tetrahydropyridin-4 yl)methylphosphinic acid (TPMPA, 15 microm). Expression of all three GABAC receptor subunits was demonstrated in the cultured neurons by RT-PCR. The ammonia treatment also activated caspases-3 and -9 as observed in immunocytochemistry for PARP p85 and western blot. Such activation of the caspases was again inhibited by CACA in a TPMPA-sensitive manner. The anti-apoptotic effect of CACA was blocked by inhibitors for MAP kinase kinase and cAMP-dependent protein kinase, PD98059 (20 microm) and KT5720 (1 microm), suggesting possible involvement of an upstream pro-apoptotic protein, BAD. Levels of phospho-BAD (Ser112 and Ser155) were decreased by the ammonia-treatment and restored by coadministration of CACA. These findings suggest that GABAC receptor stimulation protects hippocampal pyramidal neurons from ammonia-induced apoptosis by restoring Ser112- and Ser155 phospho-BAD levels. PMID- 14535962 TI - Obesity--is it a genetic disorder? AB - Obesity is one of the most pressing problems in the industrialized world. Twin, adoption and family studies have shown that genetic factors play a significant role in the pathogenesis of obesity. Rare mutations in humans and model organisms have provided insights into the pathways involved in body weight regulation. Studies of candidate genes indicate that some of the genes involved in pathways regulating energy expenditure and food intake may play a role in the predisposition to obesity. Amongst these genes, sequence variations in the adrenergic receptors, uncoupling proteins, peroxisome proliferator-activated receptor, and the leptin receptor genes are of particular relevance. Results that have been replicated in at least three genome-wide scans suggest that key genes are located on chromosomes 2p, 3q, 5p, 6p, 7q, 10p, 11q, 17p and 20q. We conclude that the currently available evidence suggests four levels of genetic determination of obesity: genetic obesity, strong genetic predisposition, slight genetic predisposition, and genetically resistant. This growing body of research may help in the development of anti-obesity agents and perhaps genetic tests to predict the risk for obesity. PMID- 14535963 TI - Issues facing clinical trials of the future. AB - Diagnostic and therapeutic technology continues to advance rapidly, as does our knowledge of therapeutics and of clinical research methods. Unfortunately, these advances have been only poorly coupled with our knowledge of therapeutic principles, leading to increasing uncertainty about which technologies are truly effective and, amongst those that are effective, which are most effective for the cost. This article presents general principles derived from several investigators' experiences with clinical trials, and uses them to suggest how future clinical trials may differ from current approaches. A proposed organization for future trials also is elucidated. PMID- 14535964 TI - Risk of haemorrhagic stroke in patients with oral anticoagulation compared with the general population. AB - OBJECTIVES: To compare the incidence of haemorrhagic stroke (HS), and the risk of fatal outcome after HS in patients with oral anticoagulation (OA) treatment and in the general population. DESIGN: Five-year cohort study. SETTING: The Anticoagulation Clinic, Malmo University Hospital, Lund, Sweden. SUBJECTS: A total of 4434 patients treated with OA (6693 treatment years) from 1 Oct 1993 to 30 Sept 1998. The population-based Stroke Register of Malmo, Lund, Sweden (STROMA). RESULTS: Forty-eight patients had HS according to ICD 9 code 430 and 431. HS occurred at a higher age in women compared with men (mean age 79.5 years vs. 74.7 years, P=0.009). The age-adjusted relative risk of HS during OA treatment was 10.9 (CI 6.7-17.6) for men and 9.3 (CI 5.7-15.0) for women, as compared with the untreated general population. Number needed to harm (NNH) (person-years) was 103 for men and 188 for women. Adjusted for age and sex, OA treatment was significantly associated with fatal outcome in patients with HS (OR=2.6, CI 1.4-4.8). CONCLUSIONS: Patients with OA treatment had approximately 10 times higher risk of HS as compared with the general population, and the risk increased markedly with age. OA treatment is associated with an increased case fatality in patients with HS. PMID- 14535965 TI - Pathophysiology of gastric acid secretion in patients with chronic renal failure: influence of Helicobacter pylori infection. AB - OBJECTIVES: The incidence of gastroduodenal diseases is high in patients with chronic renal failure (CRF). However, gastric acidity in CRF has been reported to range in level from low to high. Moreover, it remains unknown whether Helicobacter pylori infection influences gastric acidity in such patients. Thus, we aimed to clarify the pathophysiological perturbation in gastric acidity and to determine the influence of H. pylori infection in CRF. DESIGN: Case-control study. SETTING: A university hospital. SUBJECTS: Twenty-seven patients with CRF and 24 control patients, presenting with either gastrointestinal symptoms, positive faecal occult blood, or anaemia (haemoglobin <10 g dL(-1)). MEASURES: The patients underwent gastroduodenal endoscopy with simultaneous determination of H. pylori infection. Gastric ammonium concentration, serum pepsinogen I and II, and basal gastrin level were measured. Thereafter, gastric acid secretion was monitored by 24-h intragastric acidity measurement with calculation of pH-3 holding time (%) (hours showing pH>3/24 h). RESULTS: In the CRF group, pH-3 holding time of H. pylori (+) subgroup was significantly greater than that of H. pylori (-) subgroup (71.2 +/- 32.4% vs. 32.8 +/- 30.0%, mean +/- SD; P=0.03). Pepsinogen I/II ratio was inversely correlated with pH-3 holding time in the control and CRF groups. Gastric ammonium concentration in CRF/H. pylori (+) subgroup (14.1 +/- 9.2 mmol L(-1)) was significantly higher than in CRF/H. pylori (-) (2.5 +/- 2.7 mmol L(-1); P=0.002) and control/H. pylori (+) subgroups (6.1 +/ 4.2 mmol L(-1); P=0.01). Serum gastrin level was significantly higher in the CRF group than in the control group (297 +/-343 pg mL(-1) vs. 116 +/- 69 pg mL(-1); P=0.02) as a whole. However, there was no significant correlation between serum creatinine and gastrin levels in the CRF group. Gastrin level in CRF/H. pylori (+) subgroup was significantly higher than in CRF/H. pylori (-), control/H. pylori (+), and control/H. pylori (-) subgroups (423 +/-398 pg mL(-1) vs. 113 +/- 79, 124 +/- 78, and 96 +/-43 pg mL(-1), respectively; P=0.01-0.03). Significant positive correlations amongst pH-3 holding time, ammonium and gastrin concentrations were found in the CRF group, but not in the control group. CONCLUSIONS: CRF without H. pylori infection primarily shows a tendency for high gastric acidity, but without hypergastrinaemia. Persistent H. pylori infection in CRF leads to decreased acidity and, consequently, to fasting hypergastrinaemia via a feedback mechanism. The hypoacidity in CRF with H. pylori infection appears to result from neutralization of acid by ammonia as well as from gastric atrophy. Thus, H. pylori infection status critically determines perturbation in gastric acidity and fasting gastrin level in CRF. PMID- 14535966 TI - Endocrine effects of nasal continuous positive airway pressure in male patients with obstructive sleep apnoea. AB - OBJECTIVE: Obstructive sleep apnoea (OSA) is a relatively common condition producing disabling somnolence and profound physiological responses to hypoxaemic episodes during sleep, including significant oscillations in blood pressure. This study aimed to provide controlled data on the interaction between OSA and endocrine axes to establish whether overrepresentation of pathology such as hypertension and hypogonadism in OSA subjects might have an endocrine basis. DESIGN, SETTING AND SUBJECTS: Parallel randomized sham placebo controlled 1-month trial of nasal continuous positive airway pressure (nCPAP) in 101 male subjects with OSA presenting to a respiratory sleep clinic. METHODS: Analysis of gonadotrophins, testosterone, sex hormone binding protein (SHBG), prolactin, cortisol, thyroid stimulating hormone (TSH), free thyroxine (free T4), insulin like growth factor-1 (IGF-1), renin and aldosterone were performed at baseline and after 1 month's active or placebo nCPAP intervention. Quality of life questionnaire scoring was also recorded over the same time period. RESULTS: Testosterone and SHBG showed significant negative correlations with baseline OSA severity. Active treatment of OSA produced SHBG elevation and TSH reduction (P< or =0.03). Both groups showed an increase in aldosterone (P<0.001) and IGF-1 (P< or =0.03), associated with a large improvement in subjective quality of life scoring. CONCLUSIONS: These findings demonstrate significant changes in endocrine axes not previously reported in a placebo-controlled trial. OSA is a recognized reversible cause of testosterone reduction; SHBG suppression correlating to baseline OSA severity supports a diagnosis of secondary hypogonadism. Significant rises in aldosterone and IGF-1 on treatment coincide with increased physical activity and an improved quality of life score. PMID- 14535967 TI - Effects of short-term treatment with metformin on serum concentrations of homocysteine, folate and vitamin B12 in type 2 diabetes mellitus: a randomized, placebo-controlled trial. AB - OBJECTIVE: Metformin is a key treatment option in type 2 diabetes. However, metformin may decrease vitamin B12 levels and increase levels of homocysteine, a cardiovascular risk factor. We investigated whether 16 weeks of treatment with metformin affects serum concentrations of homocysteine, folate and vitamin B12 in subjects with type 2 diabetes treated with insulin. DESIGN: Placebo-controlled, randomized trial. MEASUREMENTS: at baseline and 16 weeks later. SETTING: This trial was conducted in the outpatient clinics of three general hospitals in The Netherlands. SUBJECTS: A total of 745 patients with type 2 diabetes, treated with insulin and not known with a contraindication for the use of metformin, were approached; 390 gave informed consent and entered the study. Thirty-seven subjects dropped out (12 placebo and 25 metformin users). INTERVENTION: Addition of metformin or placebo to insulin therapy. PRIMARY OUTCOME PARAMETERS: Serum homocysteine, folate, vitamin B12, indices of glycaemic control and body weight. RESULTS: Amongst those who completed 16 weeks of treatment, metformin use, as compared with placebo, was associated with an increase in homocysteine of 4% (0.2 to 8; P=0.039) and with decreases in folate [-7% (-1.4 to -13); P=0.024] and vitamin B12 [-14% (-4.2 to -24); P<0.0001]. In addition, the increase in homocysteine could be explained by the decreases in folate and vitamin B12. CONCLUSION: In patients with type 2 diabetes, 16 weeks of treatment with metformin reduces levels of folate and vitamin B12, which results in a modest increase in homocysteine. The clinical significance of these findings remains to be investigated. PMID- 14535968 TI - Effect of short-term hyperglycaemia on haemodynamics in type 1 diabetic patients. AB - OBJECTIVES: Mechanisms underlying glucose-mediated development and progression of diabetic complications are incompletely understood. We tested the impact of short term hyperglycaemia on systemic blood pressure and regulatory hormones in type 1 diabetic patients. DESIGN AND METHODS: We included 18 patients [13 men, mean (SEM) diabetes duration 10 (1) years] without signs of autonomic neuropathy or renal complications in a randomized single-blinded cross-over trial using insulin glucose clamp technique. Patients were clamped for 90 min to blood glucose of 5 mmol L(-1) (euglycaemia) and 15 mmol L(-1) (hyperglycaemia) in random order. Blood pressure was measured noninvasively every 5 min (Takeda TM2421 device). Regulatory hormones were determined at the end of each clamp period. RESULTS: Systolic blood pressure increased [mean (95% CI)] 3 (1, 5) mmHg during hyperglycaemia from 123 (SEM 2) during euglycaemia, P=0.01. Diastolic blood pressure remained unchanged at 78 (2) mmHg. Hyperglycaemia reduced plasma concentrations of: renin [14 (4, 23)%, P=0.02], angiotensin II [17 (8, 25)%, P<0.01] and adrenaline [20 (10, 29)%, P<0.01]. Plasma concentration of atrial natriuretic peptide increased by 11 (6, 17) pg mL(-1) (P<0.01) from 43 (2) pg mL( 1). We calculated a median (range) increase in extracellular volume and plasma volume (PV) of 2.6 (0.7-5.3)% and 5.0 (-4.7 to 8.6)%, respectively. CONCLUSIONS: In type 1 diabetic patients without signs of autonomic neuropathy short-term hyperglycaemia induced a modest increase in systolic blood pressure and suppression of the renin-angiotensin system, possibly caused by PV expansion because of fluid shift from intra- to extracellular compartment. PMID- 14535969 TI - Expansion of peripheral CD8+ T cells in patients with coronary artery disease: relation to cytomegalovirus infection. AB - OBJECTIVES: The nature of the immune response in coronary artery disease (CAD) is not fully defined. One pathogen that has been linked to atherogenesis, cytomegalovirus (CMV), is known to exert strong and long-lasting effects on peripheral T cells. In the present study, we investigated the effect of prior CMV infection on the immune system in CAD patients. SUBJECTS: Patients with stable angina and angiographically verified CAD (n=43) and clinically healthy controls (n=69) were included. METHODS: The expression of CD57 and CD28 on peripheral CD4+ and CD8+ T cells was evaluated with three-colour flow cytometry. The findings were related to serological markers of inflammation, T-cell activation and CMV seropositivity. RESULTS: An expansion of CD8+ T cells expressing CD57 but lacking CD28 was seen in the patient group. The numbers of CD8+ CD57+ and CD8+ CD28-T cell subsets were independently related to CMV seropositivity (P<0.001) but also to CAD per se (P<0.05). Serum concentrations of C-reactive protein (CRP) and soluble interleukin-2 receptor (sIL-2R) were elevated in the patients but not related to CMV or CD8+ T-cell subsets. CONCLUSION: A pronounced shift in peripheral T-cell homeostasis was observed in CAD patients. Primarily CMV infection but also CAD per se contributed to the expansion of CD8+ T-cell subsets. The T-cell changes were not related to a systemic inflammatory response but should rather be considered as markers of a chronic antigen exposure and/or immunosenescence in CAD. PMID- 14535970 TI - Left atrioventricular plane displacement determined by echocardiography: a clinically useful, independent predictor of mortality in patients with stable coronary artery disease. AB - BACKGROUND: Echocardiographically determined left atrioventricular plane displacement (AVPD) is strongly related to prognosis in patients with chronic heart failure and in postmyocardial infarction patients. We aimed at exploring whether AVPD, unlike ejection fraction, is related to mortality in patients with stable coronary artery disease (CAD). METHODS AND RESULTS: Atrioventricular plane displacement was assessed by two dimensionally guided M-mode echocardiography in the four and two chamber views, in 333 consecutive patients with stable CAD and an abnormal coronary angiogram. Patients were followed up for an average of 41 months. AVPD was lower in patients who died (n= 30, 9.0 %) compared with survivors (9.0 +/- 2.2 vs. 11.5 +/- 2.1 mm, P<0.0001). Amongst patients with prior myocardial infarction (n=184) AVPD was 8.7 +/- 2.3 mm in those who died (n=17) and 11.2 +/- 2.3 mm in the survivors (P<0.0001). In patients without prior myocardial infarction (n=149), AVPD was 9.4 +/- 2.1 (n=13) and 11.8 +/- 1.8 mm, respectively (P<0.0001). Age, AVPD and four other echocardiographical variables correlated significantly with prognosis in univariate logistic regression analysis. In multiple logistic regression analysis only AVPD (P<0.0001) correlated independently with mortality. CONCLUSION: Echocardiographically determined AVPDis a clinically useful, independent prognostic tool in patients with stable CAD. The presence of a documented previous myocardial infarction does not influence this observation. PMID- 14535971 TI - Corticosteroid use and risk of hip fracture: a population-based case-control study in Denmark. AB - OBJECTIVE: To examine the association between cumulative CS dose and risk of hip fracture. DESIGN: Population-based case-control design. SUBJECTS AND METHODS: A total of 6660 subjects with hip fracture and 33,272 age-matched population controls were identified using the County Hospital Discharge Registry in North Jutland County, Denmark and the Danish Central Personal Registry, respectively. Data on redeemed prescriptions for CS within the last 5 years before the index date were retrieved from a population-based prescription database, and recalculated to prednisolone equivalents. Cases and controls were categorized according to cumulative CS dose: (i) no use; (ii) <130 mg (e.g. equivalent to 30 mg of prednisolone for 4 days given for an acute exacerbation of asthma); (iii) 130-499 mg (e.g. equivalent to a short course of prednisolone of 450 mg for acute asthma); (iv) 500-1499 mg (e.g. equivalent to 7.5 mg prednisolone daily for 6 months or 800 microg day(-1) of inhaled budesonide for 1 year); and (v) > or =1500 mg (e.g. equivalent to >4.1 mg day(-1) for 1 year, a long-term high dose). Data were analysed using conditional logistic regression adjusted for potential confounders including gender, redeemed prescriptions for hormone replacement therapy, antiosteoporotic, anxiolytic, antipsychotic and antidepressant drugs. RESULTS: Compared with never users, an increased risk of hip fracture was found for CS users, with increasing cumulative doses of any type of CS use during the preceding 5 years [adjusted odds ratio (OR)=0.96, 95% confidence interval (CI)=0.89-1.04] for <130 mg prednisolone; OR=1.17 (CI=1.01-1.35) for 130-499 mg; OR=1.36 (CI=1.19-1.56) for 500-1499 mg; and OR=1.65 (CI=1.43-1.92) for > or =1500 mg. An increased risk was also found when the study population was stratified according to gender, age and type of CS (systemic or topical). CONCLUSIONS: Even a limited daily dose of CS (more than an average dose of approximately 71 microg prednisolone per day) was associated with an increased risk of hip fracture. PMID- 14535972 TI - Basal glucometabolic status has an impact on long-term prognosis following an acute myocardial infarction in non-diabetic patients. AB - OBJECTIVES: Patients with diabetes are known to have a worse prognosis after an acute myocardial infarction (AMI) compared with non-diabetic patients. The primary aim of this study was to investigate the effect of glucometabolic status on long-term prognosis in non-diabetic patients with an AMI. The second aim was to evaluate the extent to which blood glucose levels at admission depended on acute stress, assessed as serum cortisol, previous glucometabolic status, measured as haemoglobin A1c (HbA1c), or both. DESIGN: In a prospective study of patients with an AMI, blood glucose, HbA1c and cortisol were measured at admission. Fasting blood glucose was determined before discharge and also afterwards, if necessary, for classification. Patients were followed-up for 5.5 years. SUBJECTS: Of the 305 consecutive patients 24% were diagnosed as diabetic and 76% as non-diabetic. MAIN OUTCOME MEASURES: Death or non-fatal myocardial re infarction. RESULTS: In non-diabetic patients, a Cox regression model was used. With death or re-infarction as endpoint, the following prognostic factors had an impact on event-free survival: age (P<0.001), HbA1c (P=0.002), cortisol (P<0.001) and thrombolytic treatment (P=0.001). There was a correlation between cortisol and blood glucose at admission (r=0.44, P<0.001). Fasting blood glucose day 5 showed no association with event-free survival. CONCLUSIONS: In non-diabetic patients with AMI, admission HbA1c and cortisol were predictors for 5.5-year survival without recurrent non-fatal myocardial infarction. The glucometabolic status of importance for prognosis was detected by HbA1c but not by fasting blood glucose or admission blood glucose, of which the latter was influenced by cortisol. PMID- 14535974 TI - Beneficial effect of glucose-insulin-potassium infusion in noncritically ill patients has to be proven. PMID- 14535973 TI - Homocysteine and its relation to B-vitamins in Graves' disease before and after treatment: effect modification by smoking. AB - OBJECTIVES: To investigate plasma total homocysteine levels and its relation to B vitamins and smoking in Graves' disease before and after antithyroid therapy. DESIGN: A longitudinal study taking place at four hospitals in Norway. METHODS AND SUBJECTS: Plasma total homocysteine, serum folate, serum cobalamin and riboflavin, flavin mononucleotide and flavin adenine dinucleotide in plasma were investigated in 182 patients with hyperthyroidism before treatment. The same parameters were reinvestigated in 112 of these patients after attaining euthyroid state. RESULTS: In hyperthyroidism, plasma total homocysteine was low, and inversely related to folate, cobalamin and riboflavin, and positively related to serum creatinine and age. Following antithyroid therapy, total homocysteine increased and the concentration of folate, cobalamin, riboflavin, flavin mononucleotide and flavin adenine dinucleotide decreased significantly. The most pronounced reduction (35%) was observed for flavin mononucleotide. In the hyperthyroid state, smokers had lower levels of folate and flavin mononucleotide than non-smokers. After restoration of euthyroidism, both folate and riboflavin were significantly lower in smokers than non-smokers. Plasma total homocysteine increased according to decreasing quartiles of B-vitamins. For riboflavin, this relation was confined to smokers. CONCLUSION: Plasma total homocysteine changes according to thyroid status. These changes may be partly attributable to altered folate, cobalamin but also riboflavin status, particularly in smokers. PMID- 14535976 TI - Self-management of oral anticoagulation: long-term results. PMID- 14535977 TI - Nitric oxide in Helicobacter pylori-induced apoptosis and its significance in gastric carcinogenesis. PMID- 14535978 TI - Is acupuncture a risk factor for hepatitis? Systematic review of epidemiological studies. AB - OBJECTIVE: Acupuncture has been repeatedly associated with infectious hepatitis. The aim of the present systematic review was therefore to critically evaluate such data from epidemiological investigations. METHODS: Four independent literature searches were carried out to identify all epidemiological evidence linking acupuncture with hepatitis. All studies were validated by the authors and data extracted according to predefined criteria. RESULTS: Fifteen investigations fulfilled our inclusion criteria. Most studies originated from Asia. One study included markers to hepatitis A virus, three to hepatitis B virus, and 13 to hepatitis C virus. Five investigations reported associations between acupuncture and seropositivity to hepatitis C virus. In those studies, acupuncture increased the risk only modestly. CONCLUSION: A modest association between hepatitis C and acupuncture has been reported in some countries. This emphasizes the importance of exclusively using disposable acupuncture needles. PMID- 14535979 TI - Simplified 13C-urea breath test with a new infrared spectrometer for diagnosis of Helicobacter pylori infection. AB - BACKGROUND AND AIM: Infrared spectrometry has correlated excellently with mass spectrometry in detecting the ratio of 13CO(2) to 12CO(2) in breath samples. The present study aimed to evaluate the accuracy of the 13C-urea breath test (13C UBT) using a new model of infrared analyzer. METHODS: A total of 600 patients who were undergoing upper endoscopy without receiving eradication therapy were entered into the study. Culture, histology, and rapid urease test on biopsies from the antrum and corpus of the stomach were used for the determination of Helicobacter pylori infection. Breath samples were collected before and 20 min after drinking 100 mg 13C-urea in 100 mL water. The optimal cutoff value was determined by the receiver operating characteristic curve. RESULTS: Of the 586 patients who were eligible for analysis, 369 were positive for H. pylori infection, 185 were negative for H. pylori infection, and 32 were indeterminate. When the appropriate cutoff value was set at 3.5 per thousand, a sensitivity of 97.8%, a specificity of 96.8% and an accuracy of 97.5% were obtained using the 13C-UBT. The accuracy of the 13C-UBT decreased when CO(2) concentration in the breath sample was <2%, as compared with > or = 2% (93.6%vs 97.7%), mainly because of a decrease in specificity (81.8%vs 97.7%). There were 2.7% of patients with Delta13CO(2) values that fell between 3.0-4.5 per thousand, in whom the risk of error was 47%. CONCLUSIONS: The 13C-UBT performed with infrared spectrometry is a highly sensitive, specific, and non-invasive method for the detection of H. pylori infection. The immediate availability of the test result and technical simplicity make it particularly effective in routine clinical practice. PMID- 14535980 TI - Helicobacter pylori cagA, iceA and vacA status in Taiwanese patients with peptic ulcer and gastritis. AB - BACKGROUND: Helicobacter pylori causes chronic gastritis, peptic ulcer, gastric cancer and mucosa-associated lymphoid tissue (MALT) lymphoma. Different genotypes of H. pylori are confirmed from diverse geographical areas. Its association with clinical diseases remains controversial. The aim of the present study was to investigate the H. pylori vacuolating cytotoxin (vacA) alleles, cytotoxin associated gene (cagA) and iceA, in patients with peptic ulcer and gastritis. METHODS: We enrolled patients with peptic ulcer and chronic gastritis. Biopsy specimens were obtained from the antrum and lower body of the stomach. DNA extraction and polymerase chain reaction (PCR) were used to detect the presence or absence of cagA and to assess the polymorphism of vacA and iceA. RESULTS: A total of 133 patients (57 gastric ulcer, 52 duodenal ulcer, 24 chronic gastritis) had positive PCR results from biopsy specimens. Concerning genotypes, we found cagA (79% in the antrum, 92% in the body) and iceA1 (73% in the antrum, 82.8% in the body) strains in the majority of patients. The dominant vacA subtype was s1a (74.4% in the antrum, 75% in the body), followed by s1c (51.1% in the antrum, 60.5% in the body). In the middle region, the m2 strain dominated (49.6% in the antrum, 41.4% in the body), followed by m1T (19.5% in the antrum, 9.5% in the body). Mixed infection occurred in 89 patients (67%). There was no statistical difference in genotypes among the three groups. CONCLUSION: In Taiwan, H. pylori with positive cagA and iceA1 was found in the majority of cases. H. pylori with vacA s1a strains was the most common vacA subtype, followed by s1c, while s1b was rare. In the middle region, the m2 subtype was predominant followed by m1T. There was no significant association between genotypes and clinical diseases. PMID- 14535981 TI - Effect of Helicobacter pylori eradicated therapy on water gastric emptying in patients with active duodenal ulcer. AB - BACKGROUND AND AIMS: It remains debatable if duodenal ulcer (DU) or Helicobacter pylori infection has a definite impact on human gastric emptying (GE). We explored the nature of water GE in active DU patients before and after ulcer healing and the influence of H. pylori eradication on GE. METHODS: A home made applied potential tomography (APT) was used to measure liquid GE. Twelve electrodes were placed in a circular array around the upper abdomen of studied subjects. After drinking 500 mL of ion-free water, paired electrodes injected electrical current and the remaining 10 electrodes recorded signals, one-by-one in a rotating order. Based on tomographical calculation, the serial changes of averaged signals from altered resistivities were constructed to display GE. Initially, 64 H. pylori infected active DU patients were enrolled. After APT measurement, one-week triple therapy (omeprazole, amoxicillin and clarithromycin) was dispensed. Patients were asked back to determine ulcer/H. pylori status and GE on a scheduled date 3 months later. Finally, 58 patients finished the trial with valid and readable GE data obtained. RESULTS: The ulcer healing and H. pylori eradicated rates were 91.4% and 82.8%, respectively. In general, liquid GE was prolonged in all DU patients at follow up. Of 48 eradicated patients, 35.4% manifested either enhanced or delayed GE before treatment, whereas only five (10.4%) had abnormal GE after treatment (P < 0.0001). In contrast, this characteristically normalized GE was not found in non-eradicated patients. CONCLUSIONS: Water GE of active DU patients ranges from enhanced to delayed, while an effective H. pylori triple therapy is useful not only for healing ulcers, but also for restoring abnormal GE. PMID- 14535982 TI - Analysis of CYP2E1 polymorphism for the determination of genetic susceptibility to gastric cancer in Koreans. AB - BACKGROUND: Interindividual genetic differences in susceptibility to chemical carcinogens are among the most important host factors in human cancer. The present study was undertaken to reveal the association between the polymorphism of CYP2E1 (CYP2E1/PstI and CYP2E1/DraI) with genetic susceptibility to gastric cancer development in Koreans. METHODS: In the present study, 120 gastric cancer patients and 145 controls with no history of tumors were analyzed. CYP2E1 was determined by polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP), or PCR and direct gel electrophoresis. RESULTS: The overall genotype distribution of CYP2E1 was not significantly different from that of controls. However, the genotype distribution of the patient subgroups with a history of heavy cigarette smoking (>30 pack/year) in the CYP2E1/PstI and CYP2E1/DraI polymorphisms were significantly different from those of non-smoking patients (P = 0.0122 and P = 0.0029, respectively). The difference was also noticeable in the younger patient subgroup (aged /= 5 mm (P < 0.05) on radiological examination, significant regional lymph node enlargement (> 1 cm, p = 0.01) on CT scan and abrupt cut-off (P < 0.01), and separation of bile ducts (P < 0.05) on cholangiogram were differential diagnostic markers between the two groups. CONCLUSIONS: Preoperative laboratory data such as alkaline phosphatase and CA 19-9, and imaging findings such as significant wall thickening and regional lymph node enlargement on CT, abrupt cut-off and separation of bile duct on cholangiogram are useful differential diagnostic markers for benign and malignant bile duct strictures without demonstrable mass on US or CT. PMID- 14535987 TI - Expression of hepatoma-derived growth factor in hepatocarcinogenesis. AB - BACKGROUND AND AIM: The present study investigated the expression of hepatoma derived growth factor (HDGF) in human hepatocellular carcinoma (HCC) and in the liver during hepatocarcinogenesis in two rodent models. METHODS: Expression of HDGF was analyzed using northern blotting and immunohistochemistry in the human and rodent models. RESULTS: Hepatoma-derived growth factor was more highly expressed in HCC than in the adjacent liver in humans with hepatitis, as shown by northern blotting. Using immunohistochemistry with the specific anti-HDGF antibody, HDGF was more strongly and frequently expressed in the nucleus and cytoplasm of HCC cells than in the adjacent normal hepatocytes. Hepatoma-derived growth factor was also more strongly expressed in the tumors than in the adjacent fatty liver of fatty liver Shionogi (FLS) mice, than in the cirrhotic liver of choline-deficient amino acid feeding rats, as shown by northern blotting and immunohistochemistry. In the liver of FLS mice, HDGF expression increased gradually from the age of 24 weeks through to 52 weeks after birth, showing that HDGF expression was already increased at an early stage before tumor development. In the non-tumorous liver with fatty change, the foci expressing HDGF appeared at 24 weeks of age, which were the activated macrophage clusters with enhanced DNA synthesis and fat droplets. It is suggested that HDGF was secreted or released from these foci and stimulated hepatocyte proliferation in a paracrine manner in FLS mice, and stimulated the proliferation of hepatic tumor cells in an autocrine manner. CONCLUSIONS: The present findings suggest that HDGF plays an important role in the development or progression of HCC in humans and rodents. PMID- 14535989 TI - Possible involvement of tetraspanin CO-029 in hematogenous intrahepatic metastasis of liver cancer cells. AB - BACKGROUND AND AIM: A correlation between overexpression of tetraspanin CO-029 and the intrahepatic spread of hepatocellular carcinoma (HCC) has been observed in surgically resected specimens from humans. However, the cellular mechanisms involved in CO-029 protein modulation of the metastatic phenotype are unknown. In the present study, CO-029 cDNA was stably transfected into a non-metastatic human HCC cell line to investigate whether it could directly promote metastasis. METHODS: We constructed a human HCC cell line that stably overexpressed CO-029 and mock transfectants. Using these transfectants, we examined cell proliferation characteristics in monolayer culture and the ability to adhere to culture plates coated with laminin, fibronectin, vitronectin and collagen type-I or type-IV. Orthotopic implantation of these transfectants to SCID mice was also performed. RESULTS: Several clones of CO-029 transfectants and mock transfectants were established. The growth rates and adhesive properties to the extracellular matrix did not differ between CO-029 and mock transfectants. When orthotopically implanted, the size of the primary tumor in the liver did not differ between CO 029 and the mock transfectants. However, only CO-029 positive clones developed intrahepatic metastatic lesions. CONCLUSION: These results suggest that CO-029 might be involved in hematogenous intrahepatic metastasis, although the precise cellular mechanisms involved remain unknown. PMID- 14535988 TI - Parenchymal transforming growth factor beta-1: its type II receptor and Smad signaling pathway correlate with inflammation and fibrosis in chronic liver disease of viral etiology. AB - BACKGROUND AND AIMS: Transforming growth factor-beta (TGF-beta) system is involved in the control of cell growth and extracellular matrix formation. Previous studies in patients with chronic liver disease have shown that increased TGF-beta expression significantly correlates with the degree of hepatic fibrosis. The aim of our study was to define TGF-beta system expression in hepatic parenchymal cells and its significance in patients with differing extents of chronic liver disease of viral etiology. METHODS: Expression of TGF-beta 1, TGF beta 1 type II receptor (TGF-beta RII) and the Smad signaling pathway was evaluated in consecutive liver sections of 77 patients with chronic liver disease (65 HCV positive and 12 HBV positive). Results were correlated with histological scores and apoptotic activity. RESULTS: TGF-beta 1 was demonstrated in the liver of 30/56 (53.6%) patients with chronic hepatitis and 20/21 (95%) patients with cirrhosis, but in none of the 20 normal livers. Positive cytokine reaction was seen both in stromal cells and hepatocytes. Expression of TGF-beta RII and Smad proteins showed a distribution pattern similar to that of TGF-beta, with a direct correlation in terms of immunoreactivity extent. A significant correlation was found between parenchymal expression of TGF-beta system and inflammatory and fibrosis scores. No correlation was found with apoptotic index and other morphological, clinical or virological parameters. CONCLUSIONS: The TGF-beta system is up-regulated at the ligand, receptor and signaling level in the liver of patients with more active disease. The strong expression of TGF-beta and the Smad pathway in parenchymal cells suggests that hepatocytes, in addition to mesenchymal cells, may play an important role in the progression of liver disease. PMID- 14535991 TI - Hepatobiliary and pancreatic: cholangiocellular cancer and hepatitis C. PMID- 14535990 TI - Elevation of serum and ascites cancer antigen 125 levels in patients with liver cirrhosis. AB - BACKGROUND AND AIM: Serum cancer antigen (CA) 125 elevation has been reported in patients with liver disease, but it is poorly characterized. The present study aimed to evaluate the range of serum and ascitic CA 125 levels in patients with liver cirrhosis and to explore possible factors associated with CA 125 elevation. METHODS: A total of 70 patients were studied. Group I consisted of 30 patients with liver cirrhosis with or without ascites. Group II consisted of 30 patients with digestive malignant tumors with or without ascites. Group III consisted of 10 patients with benign ascites. The CA 125 levels were measured in the serum of all patients and also simultaneously in the ascitic fluid of 15 patients. RESULTS: Serum CA125 levels in 80% of (24/30) patients from group I were elevated, particularly in those with ascites, irrespective of the etiology of cirrhosis. Serum CA 125 levels were correlated with Child-Pugh scores (r = 0.38), but not significantly (P = 0.06). All patients from group II with ascites and from group III had elevated serum CA 125 levels, but there was no difference in the serum CA 125 levels between patients with ascites from group I (275 +/- 175 U/mL), group II (368 +/- 190 U/mL) or group III (396 +/- 287 U/mL), nor was there a significant difference in ascitic CA 125 levels (P > 0.05). The levels of serum CA 125 (198 +/- 108 U/mL) were lower than, but correlated with that of ascites (460 +/- 234 U/mL, r = 0.58, P = 0.026). The elevation of serum CA 125 accompanied by abnormalities of other tumor markers was more common in malignant ascites than in benign ascites (90% compared with 6%, P < 0.05). CONCLUSION: The elevation of serum CA 125 is common in patients with liver cirrhosis. It is related to the presence of ascites, and possibly to the insufficiency of liver function, but not the etiology of cirrhosis and ascites. Serum CA 125 probably comes from ascites. It usually predicts benign disease if the elevation of serum or ascites CA 125 is not accompanied by the abnormalities of other tumor markers. PMID- 14535992 TI - Gastrointestinal: esophageal intramural hematoma. PMID- 14535993 TI - Coexistence of focal nodular hyperplasia and pseudotumor caused by focal spared lesion in the liver of a young man. AB - We report a case of 19-year-old male with an underlying case of chronic hepatitis C infection who suffered from two types of benign liver tumor: focal nodular hyperplasia (FNH) and pseudotumor due to focal fatty spared area in a fatty liver. These two kinds of lesions rarely occur simultaneously. The spectral Doppler ultrasound (US) images of these lesions were also provided. We suggest that spectral Doppler US provides an alternative diagnostic tool for the differentiation of liver tumors regarding their vascular pattern, which might help ensure a correct diagnosis. PMID- 14535994 TI - Case of intraductal papillary mucinous tumor in which endosonography-guided fine needle aspiration biopsy caused dissemination. PMID- 14535995 TI - Autoimmune hepatitis associated with celiac disease in childhood: report of two cases. PMID- 14535996 TI - Interactions between inducible nitric oxide and other inflammatory mediators during Helicobacter pylori infection. AB - BACKGROUND: Recent studies in both humans and animal models strongly suggest the contribution of the host immune response to Helicobacter pylori-related disease. Inducible nitric oxide synthase has been shown to be up-regulated in the gastric epithelium during H. pylori gastritis, suggesting a role in inflammation. MATERIALS AND METHODS: C57BL/6 wild-type and inducible nitric oxide synthase gene knockout mice were infected with H. pylori strain SS1. Expression of macrophage inflammatory protein-2 (MIP-2), interleukin-1 beta (IL-1 beta), Th1 (IL-2 and gamma interferon) and Th2 (IL-4 and IL-10) cytokines, and inducible cyclooxygenase mRNA in mice was determined in mouse gastric tissues and quantified using either competitive reverse transcription-polymerase chain reaction or competitive polymerase chain reaction following reverse transcription. RESULTS: The Th1 cytokine gamma interferon was only detected in wild-type and inducible nitric oxide synthase gene knockout infected mice, while a Th2 (IL-4) response was not detected. H. pylori induced MIP-2 and IL-1 beta mRNA in mice. CONCLUSIONS: Because similar levels of inflammatory mediators were noted in both wild-type and nitric oxide synthase gene knockout infected mice, our data suggest that inducible nitric oxide synthase does not influence expression of these inflammatory mediators in the early stages of H. pylori infection in mice. PMID- 14535997 TI - Oxidative damage of the gastric mucosa in Helicobacter pylori positive chronic atrophic and nonatrophic gastritis, before and after eradication. AB - BACKGROUND: Helicobacter pylori is the main cause of gastritis and a primary carcinogen. The aim of this study was to assess oxidative damage in mucosal compartments of gastric mucosa in H. pylori positive and negative atrophic and nonatrophic gastritis. MATERIALS AND METHODS: Five groups of 10 patients each were identified according to H. pylori positive or negative chronic atrophic (Hp CAG and CAG, respectively) and nonatrophic gastritis (Hp-CG and CG, respectively), and H. pylori negative normal mucosa (controls). Oxidative damage was evaluated by nitrotyrosine immunohistochemistry in the whole mucosa and in each compartment at baseline and at 2 and 12 months after eradication. Types of intestinal metaplasia were classified by histochemistry. RESULTS: Total nitrotyrosine levels appeared significantly higher in H. pylori positive than in negative patients, and in Hp-CAG than in Hp-CG (p <.001); no differences were found between H. pylori negative gastritis and normal mucosa. Nitrotyrosine were found in foveolae and intestinal metaplasia only in Hp-CAG. At 12 months after H. pylori eradication, total nitrotyrosine levels showed a trend toward a decrease in Hp-CG and decreased significantly in Hp-CAG (p =.002), disappearing from the foveolae (p =.002), but remaining unchanged in intestinal metaplasia. Type I and II of intestinal metaplasia were present with the same prevalence in Hp-CAG and CAG, and did not change after H. pylori eradication. CONCLUSIONS: Oxidative damage of the gastric mucosa increases from Hp-CG to Hp-CAG, involving the foveolae and intestinal metaplasia. H. pylori eradication induces a complete healing of foveolae but not of intestinal metaplasia, reducing the overall oxidative damage in the mucosa. PMID- 14535998 TI - Induction of cyclooxygenase 2 by Escherichia coli but not Helicobacter pylori lipopolysaccharide in gastric epithelial cells in vitro. AB - BACKGROUND: Cyclooxygenase 2 (COX-2) is an inducible enzyme that plays a key role in the synthesis of prostaglandins in response to inflammatory stimuli. It is expressed in the gastric mucosa as part of the response to infection with Helicobacter pylori. The specific interaction between H. pylori and the gastric epithelium that results in COX-2 expression has not been identified. METHODS: In order to investigate the hypothesis that lipopolysaccharide (LPS) from H. pylori plays a role in the induction of cyclooxygenase 2 in the stomach, gastric cell lines MKN-7 and MKN-45 were incubated with LPS from either H. pylori NCTC 11637 or Escherichia coli 055:B5. Incubation of cells with live H. pylori NCTC 11637 was also carried out as a positive control. Cells were then analysed for COX-2 protein and mRNA and prostaglandin E2 synthesis. RESULTS: Cyclooxygenase 2 protein and mRNA expression was induced by E. coli LPS and live H. pylori, but not by H. pylori LPS. Prostaglandin E2 synthesis increased in a dose-dependent manner in both cell lines with E. coli but not H. pylori LPS. CONCLUSIONS: H. pylori LPS is of low biological activity when compared with E. coli LPS in its ability to induce the expression of cyclooxygenase 2 and synthesis of prostaglandin E2. This may provide one mechanism by which H. pylori minimizes the inflammatory response in the gastric mucosa, allowing chronic infection. PMID- 14535999 TI - The MUC5AC glycoprotein is the primary receptor for Helicobacter pylori in the human stomach. AB - BACKGROUND AND OBJECTIVES: Helicobacter pylori shows a characteristic tropism for the mucus-producing gastric epithelium. In infected patients, H. pylori colocalizes in situ with the gastric secretory mucin MUC5AC. The carbohydrate blood-group antigen Lewis B (LeB) was deemed responsible for the adherence of H. pylori to the gastric surface epithelium. We sought to determine if MUC5AC is the carrier of LeB, and thus if MUC5AC is the underlying gene product functioning as the main receptor for H. pylori in the stomach. METHODS: We studied three types of human tissue producing MUC5AC: Barrett's esophagus (BE), normal gastric tissue, and gastric metaplasia of the duodenum (GMD). Tissue sections were immuno fluorescently stained for MUC5AC or LeB, and subsequently incubated with one of three strains of Texas red-labeled H. pylori, one of which was unable to bind to LeB. We determined the colocalization of MUC5AC or LeB with adherent H. pylori. RESULTS: The binding patterns for the two LeB-binding strains to all tissues were similar, whereas the strain unable to bind to LeB did not bind to any of the tissues. In normal gastric tissue, the LeB-binding strains always bound to MUC5AC and LeB-positive epithelial cells. In four nonsecretor patients, colocalization of the LeB-binding strains was found to MUC5AC-positive gastric epithelial cells. In BE, the LeB-binding H. pylori strains colocalized very specifically to MUC5AC positive cells. MUC5AC-producing cells in GMD contained LeB. Yet, LeB-binding H. pylori not only colocalized to MUC5AC or LeB present in GMD, but also bound to the LeB-positive brush border of normal duodenal epithelium. CONCLUSIONS: Mucin MUC5AC is the most important carrier of the LeB carbohydrate structure in normal gastric tissue and forms the major receptor for H. pylori. PMID- 14536000 TI - Helicobacter-induced gastritis in mice not expressing metallothionein-I and II. AB - BACKGROUND: Helicobacter pylori a primary cause of gastritis and peptic ulcer disease, is associated with increased production of reactive oxygen species within the gastric mucosa. Metallothionein (MT), a low-molecular-weight, cysteine rich, metal-binding ligand, has been shown to sequester reactive oxygen species and reduce tissue damage. This study investigates the role of MT in H. pylori induced gastritis in mice. MATERIALS AND METHODS: Control (MT+/+) and MT-null (MT /-) mice were inoculated with either 1 x 108H. pylori or H. felis, and were infected for 4, 8 and 16 weeks or 8 weeks, respectively. H. pylori load was determined by culture. Myloperoxidase activity and MT levels were also determined. RESULTS: The stomachs of H. felis-infected mice were more severely inflamed than those of H. pylori-infected mice. H. felis-induced gastritis was more severe (p =.003) in MT-/- than in MT+/+ mice. MT-/- mice also had higher (60%; p <.05) H. pylori loads than MT+/+ mice 4 weeks after infection but not 8 or 16 weeks after infection. Myloperoxidase activity with H. pylori was similar between MT+/+ and MT-/- mice. Thirty-three per cent greater (p <.05) myloperoxidase activity was observed in MT-/- than in MT+/+ mice infected with H. felis. In MT+/+ mice infected with H. pylori, liver MT was increased by 33 and 39% (p <.05) at 8 and 16 weeks, respectively, whereas gastric MT increased by 46% (p <.05) at 4 weeks and declined to baseline levels at 8 and 16 weeks. CONCLUSIONS: Mice lacking MT are more susceptible to H. pylori colonization and gastric inflammation, indicating that MT may be protective against H. pylori induced gastritis. PMID- 14536001 TI - Inhibition of Helicobacter pylori-induced nuclear factor-kappa B activation and interleukin-8 gene expression by ecabet sodium in gastric epithelial cells. AB - BACKGROUND: Helicobacter pylori stimulates nuclear factor-kappa B (NF-kappa B) activation and chemokine interleukin-8 (IL-8) expression in gastric epithelial cells. Ecabet sodium (ecabet), a locally acting antiulcer drug, is known to have anti-H. pylori activity. However, there is little understanding of how ecabet induces anti-inflammatory activity in gastric epithelial cells infected with H. pylori. The aim of this study was to investigate the effects of ecabet on IL-8 gene expression and NF-kappa B activation in human gastric epithelial cells infected with H. pylori. MATERIALS AND METHODS: After Hs746T, MKN-45, or SNU-5 gastric epithelial cell lines had been infected with cagA+cytotoxin+H. pylori in the presence of ecabet, IL-8 mRNA expression was assessed by quantitative reverse transcription-polymerase chain reaction, and IL-8 secretion was measured by enzyme-linked immunosorbent assay. NF-kappa B and inhibitory kappa B-alpha (I kappa B alpha) signals were assayed by electrophoretic mobility shift assay and Western blot, respectively. The activation of NF-kappa B and IL-8 reporter genes was determined by luciferase assay. RESULTS: Ecabet showed no antimicrobial activiy against Gram-positive or -negative bacteria. However, ecabet inhibited transcription of the IL-8 gene and secretion of IL-8 by gastric epithelial cells infected with H. pylori at a concentration of 5 micro g/ml. Moreover, ecabet inhibited the activation of NF- kappa B and the degradation of I kappa B alpha in gastric epithelial cells in response to H. pylori infection. In addition, the NF kappa B signal inhibited by ecabet was comprised predominantly of heterodimers of p65/p50. CONCLUSIONS: Ecabet inhibited H. pylori-induced IL-8 gene transcription and secretion by suppressing the NF-kappa B signal. This inhibition might be one pathway by which ecabet exerts its anti-inflammatory effect on H. pylori-induced gastric inflammation. PMID- 14536002 TI - Intensity of inflammation, density of colonization and interleukin-8 response in the gastric mucosa of children infected with Helicobacter pylori. AB - BACKGROUND: Few reports exist on inflammation and interleukin (IL)-8 response in H. pylori-infected children. The aim of this study was to determine the intensity of inflammation, density of colonization and magnitude of IL-8 response in children with and without H. pylori infection. MATERIALS AND METHODS: We studied 45 children with dyspeptic symptoms, 21 infected with H. pylori and 24 without infection. Antrum and corpus gastric biopsies were obtained and studied for H. pylori infection with an immunofluorescence technique and for IL-8 with an immunohistochemical assay. Biopsy specimens were stained with hematoxilin and eosin and gastritis was graded according to the Sydney system. The magnitudes of the IL-8 response and H. pylori colonization were estimated microscopically with image analyzer software. RESULTS: In H. pylori-infected children, mild mono nuclear cell infiltration was found in 50%, and no neutrophils in 40% of cases. In the antrum but not in the corpus, the intensity of colonization correlated with neutrophil and mononuclear cell infiltration. The IL-8 response was significantly higher in the antrum (p <.05) and corpus (p <.02) of infected children, and was localized mainly in the surface and crypts of the epithelium. No correlation was found between the magnitude of the IL-8 response and the infiltration of either neutrophil or mononuclear cells. CONCLUSIONS: In H. pylori infected children, poor mononuclear and neutrophil infiltration was observed. Infection was associated with a higher IL-8 response by gastric epithelial cells. The density of colonization but not the IL-8 response correlated with neutrophil cell infiltration. PMID- 14536003 TI - A protocol for isolating putative Helicobacter pylori from fecal specimens and genotyping using vacA alleles. AB - BACKGROUND: This study outlines steps for isolating and culturing Helicobacter pylori from freshly voided fecal specimens and genotyping isolates for vacA alleles. MATERIALS AND METHODS: A family with four H. pylori-infected members participated in this pilot study. Criterion for participation was a positive test for H. pylori by the urea breath test. Fecal specimens from children were taken from a freshly soiled diaper, placed in cold buffer, and prepared for culture in less than 2 hours. Culturing of H. pylori utilized selective culture media and isolates were screened for negative Gram stain, positive catalase and oxidase tests, and positive H. pylori 16S ribosomal RNA polymerase chain reaction (PCR). Strain types were determined by vacA genotyping. RESULTS: The isolation procedure is relatively simple, although 5-7 days are required for H. pylori culturing. Isolation and purification of DNA eliminated PCR inhibitors and resulted in reliable analyses. All four family members were infected with the same H. pylori strain with a genotype of vacA s1a/m2. CONCLUSION: This research lays the foundation for developing a routine and direct noninvasive method to detect the presence of H. pylori in fecal specimens. It is especially convenient for diagnosing children and infants, as samples can be obtained from soiled diapers. Culturing H. pylori from fecal samples in certain cases is important for antibiotic resistant studies prior to treating infected patients and for strain genotyping in epidemiological studies to determine transmission. PMID- 14536004 TI - Placental acquisition of maternal specific IgG and Helicobacter pylori colonization in infancy. AB - BACKGROUND: Colonization with Helicobacter pylori generally occurs in infancy, and the microorganism is often acquired from close family members. Rate of infant colonization may be affected by maternal immune status. METHODS: To investigate the potential protective effect of anti-H. pylori immunoglobulin G (IgG) acquired via the placenta, 65 mothers and their infants were studied from the infant's birth for 1 year. Circulating IgG antibodies were measured by enzyme-linked immunosorbent assay (ELISA) in cord blood and every 8 weeks. Immunoblotting was performed on sera from infants with significant increases in IgG levels. Rate of infant H. pylori colonization was measured by 13C urea breath tests every 4 weeks from the age of 12 weeks. RESULTS: Maternal and infant cord blood specific IgG levels were correlated (R2 =.747, p <.001). Infant H. pylori specific IgG fell 5 fold compared to maternal levels over the first 6 months of life, and rose subsequently in many cases, with the development of novel immunoblot patterns. There were no significant associations between the age at first positive urea breath test and maternal or infant cord specific H. pylori IgG levels. CONCLUSIONS: Transplacentally acquired specific IgG antibody does not protect infants from colonization by H. pylori. PMID- 14536005 TI - Effect of angular error on tissue Doppler velocities and strain. AB - One of the major criticisms of ultrasound Doppler is its angle dependency, that is its ability to measure velocity components directly to or from the transducer only. The present article aims to investigate the impact of this angular error in a clinical setting. Apical two- and four-chamber views were recorded in 43 individuals, and the myocardium was marked by hand in each image. We assume that the main direction of the myocardial velocities is longitudinal and correct for the angular error by backprojecting measured velocities onto the longitudinal direction drawn. Strain was calculated from both corrected and uncorrected velocities in 12 segments for each individual. The results indicate that the difference between strain values calculated from corrected and uncorrected velocities is insignificant in 5 segments and within a decimal range in 11 segments. The biggest difference between measured and corrected strain values was found in the apical segments. Strain is also found to be more robust against angular error than velocities because the difference between corrected and uncorrected values is smaller for strain. Considering that there are multiple sources of noise in ultrasound Doppler measurements, the authors conclude that the angular error has so little impact on longitudinal strain that correction for this error can safely be omitted. (ECHOCARDIOGRAPHY, Volume 20, October 2003) PMID- 14536006 TI - Necrotizing vasculitis: a cause of aortic insufficiency and conduction system disturbance. AB - Cardiac involvement in vasculitis syndromes is uncommon. We describe a 50-year old male who presented with progressive dyspnea and myalgies. Echocardiogram revealed significant thickening of aortic root, aortic cusps, and anterior mitral valve leaflet, with severe aortic regurgitation that required aortic valve replacement. Furthermore, this patient suffered progressive atrioventricular block that needed implantation of a pacemaker. The study performed disclosed the presence of necrotizing vasculitis positive for perinuclear antineutrophil cytoplasmic antibody. PMID- 14536007 TI - Live three-dimensional echocardiography: imaging principles and clinical application. AB - Live three-dimensional echocardiography (L3DE) is an important breakthrough in the field of medical ultrasound. It will provide a great potential tool for clinical diagnosis and treatment. In this article, the authors first review the bottlenecks in 3D cardiac imaging and the technical principles of L3DE that have been used to overcome some of these problems. We then discuss the scanning methods, clinical usefulness, and the future of L3DE, drawing on our experiences in examining 124 human patients and in conducting animal verification studies with a live 3D ultrasound system. PMID- 14536008 TI - Mitral and aortic valve thickening associated with galactosialidosis: echocardiographic features of a lysosomal storage disease. PMID- 14536010 TI - Transesophageal echocardiographic identification of normal and stenosed posterior intercostal arteries. AB - We describe our experience in using transesophageal echocardiography to identify both right and left posterior intercostal arteries and assessing them for stenosis. PMID- 14536009 TI - Diastolic flutter of the ventricular septum due to bioprosthetic mitral valve stenosis, without aortic regurgitation. PMID- 14536011 TI - Live three-dimensional transthoracic echocardiographic identification of discrete subaortic membranous stenosis. AB - We describe an adult patient in whom live three-dimensional transthoracic echocardiography illustrated the exact site and full extent of the subaortic membrane as well as the narrow opening within the membrane, indicative of severe stenosis. To our knowledge this has not been reported previously. PMID- 14536013 TI - Real-time three-dimensional echocardiography using a novel matrix array transducer. AB - Three-dimensional echocardiography has multiple advantages over two-dimensional echocardiography, such as accurate left ventricular quantification and improved spatial relationships. However, clinical use of three-dimensional echocardiography has been impeded by tedious and time-consuming methods for data acquisition and post-processing. A newly developed matrix array probe, which allows real-time three-dimensional imaging with instantaneous on-line volume rendered reconstruction, direct manipulation of thresholding, and cut planes on the ultrasound unit may overcome the aforementioned limitations. This report will review current methods of three-dimensional data acquisition, emphasizing the real-time methods and clinical applications of the new matrix array probe. PMID- 14536014 TI - Use of contrast enhancement for the assessment of left ventricular function. AB - Endocardial visualization is essential for accurate interpretation of regional wall thickening abnormalities, which constitute the diagnostic hallmark of coronary artery disease, and for reproducible assessment of left ventricular (LV) ejection fraction. Unfortunately, in many cardiac patients, poor image quality does not allow consistent visualization, let alone automated detection, of the endocardial boundary in all segments throughout the cardiac cycle. Continuing efforts directed towards improving endocardial visualization made by both echocardiographic imaging equipment manufacturers and researchers have recently produced new objective, quantitative approaches for the assessment of global and regional LV function. Although these developments have not been integrated into routine clinical practice, they may have a positive impact on endocardial visualization and, thus, eventually improve the ability to accurately and objectively evaluate LV performance. In this review, we discuss some of the latest developments in the use of contrast for the evaluation of LV function. PMID- 14536015 TI - Interpretation of left ventricular wall motion during stress testing. AB - This article describes the obstacles to stress echocardiographic interpretation, and reviews the techniques currently available that offer a more objective approach to stress wall motion analysis than the conventional visual methodology. These techniques include Doppler-based methods, such as myocardial Doppler velocity and strain rate imaging, as well as automated border detection techniques, such as acoustic quantification and color kinesis. PMID- 14536016 TI - New techniques for the assessment of regional left ventricular wall motion. AB - The assessment of regional left ventricular (LV) function has been an important yet unresolved problem since the introduction of echocardiography as a diagnostic tool. Abnormal regional LV wall motion is an early finding in multiple cardiac pathologies and its diagnosis is of critical importance. In the last few years diagnostic procedures based on combined use of existing echocardiographic technologies were geared toward improving the accuracy of detection of baseline and/or induced regional wall motion abnormalities. One of the assumptions is that the combination of reduced LV wall thickening and reduced myocardial velocities can be used to accurately diagnose regional myocardial dysfunction. In this article we will discuss several new techniques for the quantification of regional LV function using Doppler echocardiography. PMID- 14536017 TI - Assessment of global and regional left ventricular diastolic function in hypertensive heart disease using automated border detection techniques. AB - Acoustic quantification (AQ) and color kinesis (CK) are techniques that involve automated detection and tracking of endocardial borders. These methods are useful for the evaluation of global and regional left ventricular (LV) systolic function and more recently have been applied to evaluating LV diastolic performance. Assessment of diastolic dysfunction in hypertensive heart disease is a relevant clinical issue in which these techniques have proven useful. The diastolic portion of left atrium and LV AQ area waveforms are frequently abnormal in patients with left ventricular hypertrophy (LVH). Left ventricular AQ curves consistently demonstrate reduced rapid filling fraction (RFF) and peak rapid filling rate (PRFR), elevated atrial filling fraction (AFF), peak atrial filling rate (PAFR), and reductions in the ratio PRFR/PAFR. Acoustic quantification complements traditional Doppler echocardiographic evaluation of global diastolic function. Many patients with significant LVH and normal Doppler diastolic parameters can be identified as having diastolic dysfunction with AQ. In addition, CK has allowed the evaluation of regional diastolic performance in hypertensive patients. Regional filling curves obtained from CK have demonstrated that endocardial diastolic motion is commonly delayed and heterogeneous in patients with LVH. PMID- 14536018 TI - Three-dimensional transesophageal echocardiographic demonstration of intraatrial baffle obstruction. AB - We report an adult patient with transposition of the great arteries status post Mustard procedure in whom three-dimensional transesophageal echocardiography demonstrated intraatrial baffle obstruction. The baffle could be visualized in both long-axis and "en face" short-axis views. PMID- 14536021 TI - Is undergraduate physiotherapy study a risk factor for low back pain? A prevalence study of LBP in physiotherapy students. AB - BACKGROUND: Following reports of high prevalence of low back pain (LBP) in young physiotherapists, we investigated whether LBP was a problem for undergraduate physiotherapy students. METHOD: Physiotherapy students enrolled in one Australian tertiary institution completed a validated self-administered questionnaire in April 2001, seeking information on LBP prevalence (lifetime, 12 month, one-month, one-week), and its risk factors. The survey incorporated the Nordic back questionnaire, questions on common risks for LBP, and purpose-built questions regarding educational exposures. Univariate logistic regression models were applied to test associations. RESULTS AND DISCUSSION: 72% students responded. LBP prevalence was 69% (lifetime), 63% (12-month), 44% (one-month), 28% (one-week). The risk of LBP increased significantly for students once they completed first year. Being aged 20 or 21 years (final year students) was significantly associated with all measures of LBP, compared with the youngest students. Exposure to tertiary study of greater than two years was associated with lifetime, 12 month and one-month LBP prevalence. Spending more than 20 hours in the past month 'sitting looking down' was significantly associated with one-month LBP prevalence. Similar exposure to 'treating patients' was significantly associated with one-month and one-week LBP prevalence. CONCLUSIONS: Physiotherapy students should be alerted to the likelihood of LBP and is potential causes during their training, so that they enter the workforce with reduced risk of LBP. The potential for other undergraduate students to suffer LBP should also be considered. PMID- 14536022 TI - Serial pathways from primate prefrontal cortex to autonomic areas may influence emotional expression. AB - BACKGROUND: Experiencing emotions engages high-order orbitofrontal and medial prefrontal areas, and expressing emotions involves low-level autonomic structures and peripheral organs. How is information from the cortex transmitted to the periphery? We used two parallel approaches to map simultaneously multiple pathways to determine if hypothalamic autonomic centres are a key link for orbitofrontal areas and medial prefrontal areas, which have been associated with emotional processes, as well as low-level spinal and brainstem autonomic structures. The latter innervate peripheral autonomic organs, whose activity is markedly increased during emotional arousal. RESULTS: We first determined if pathways linking the orbitofrontal cortex with the hypothalamus overlapped with projection neurons directed to the intermediolateral column of the spinal cord, with the aid of neural tracers injected in these disparate structures. We found that axons from orbitofrontal and medial prefrontal cortices converged in the hypothalamus with neurons projecting to brainstem and spinal autonomic centers, linking the highest with the lowest levels of the neuraxis. Using a parallel approach, we injected bidirectional tracers in the lateral hypothalamic area, an autonomic center, to label simultaneously cortical pathways leading to the hypothalamus, as well as hypothalamic axons projecting to low-level brainstem and spinal autonomic centers. We found densely distributed projection neurons in medial prefrontal and orbitofrontal cortices leading to the hypothalamus, as well as hypothalamic axonal terminations in several brainstem structures and the intermediolateral column of the spinal cord, which innervate peripheral autonomic organs. We then provided direct evidence that axons from medial prefrontal cortex synapse with hypothalamic neurons, terminating as large boutons, comparable in size to the highly efficient thalamocortical system. The interlinked orbitofrontal, medial prefrontal areas and hypothalamic autonomic centers were also connected with the amygdala. CONCLUSIONS: Descending pathways from orbitofrontal and medial prefrontal cortices, which are also linked with the amygdala, provide the means for speedy influence of the prefrontal cortex on the autonomic system, in processes underlying appreciation and expression of emotions. PMID- 14536024 TI - Distinction between cold-sensitive and -tolerant jute by DNA polymorphisms. AB - Jute is the principal coarse fiber for commercial production and use in Bangladesh. Therefore, the development of a high-yielding and environmental stress tolerant jute variety would be beneficial for the agro economy of Bangladesh. Two molecular fingerprinting techniques, random-amplified polymorphic DNA (RAPD) and amplified-fragment length polymorphism (AFLP) were applied on six jute samples. Two of them were cold-sensitive varieties and the remaining four were cold-tolerant accessions. RAPD and AFLP fingerprints were employed to generate polymorphism between the cold-sensitive varieties and cold-tolerant accessions because of their simplicity, and also because there is no available sequence information on jute. RAPD data were obtained by using 30 arbitrary oligonucleotide primers. Five primers were found to give polymorphism between the varieties that were tested. AFLP fingerprints were generated using 25 combinations of selective-amplification primers. Eight primer combinations gave the best results with 93 polymorphic fragments, and they were able to discriminate the two cold-sensitive and four cold-tolerant jute populations. A cluster analysis, based on the RAPD and AFLP fingerprint data, showed the population-specific grouping of individuals. This information could be useful later in marker-aided selection between the cold-sensitive varieties and cold tolerant jute accessions. PMID- 14536023 TI - Domain analysis of the tubulin cofactor system: a model for tubulin folding and dimerization. AB - BACKGROUND: The correct folding and dimerization of tubulins, before their addition to the microtubular structure, needs a group of conserved proteins called cofactors A to E. The biochemical analysis of cofactors gave an insight to their general functions, however not much is known about the domain structure and detailed, molecular function of these proteins. RESULTS: Combining modelling and fold prediction tools, we present 3D models of all cofactors, including several previously unannotated domains of cofactors B-E. Apart from the new HEAT and Armadillo domains in cofactor D and an unusual spectrin-like domain in cofactor C, we have identified a new subfamily of ubiquitin-like domains in cofactors B and E. Together, these observations provide a reliable, molecular level model of cofactor complex. CONCLUSION: Distant homology searches allowed the identification of unknown regions of cofactors as self-reliant domains and allow us to present a detailed hypothesis of how a cofactor complex performs its function. PMID- 14536025 TI - Characterization of a salicylic acid- and pathogen-induced lipase-like gene in Chinese cabbage. AB - A cDNA clone for a salicylic acid-induced gene in Chinese cabbage (Brassica rapa subsp. pekinensis) was isolated and characterized. The cabbage gene, designated Br-sil1 (for Brassica rapa salicylate-induced lipase-like 1 gene), encodes a putative lipase that has the family II lipase motif GDSxxDxG around the active site serine. A database search showed that plant genomes have a large number of genes that contain the family II lipase motif. The lipase-like proteins include a myrosinase-associated protein, an anther-specific proline-rich protein APG, a pollen coat protein EXL, and an early nodule-specific protein. The Br-sil1 gene is strongly induced by salicylic acid and a nonhost pathogen, Pseudomonas syringae pv. tomato, that elicits a hypersensitive response in Chinese cabbage. Treatment of the cabbage leaves with BTH, methyl jasmonate, or ethephon showed that the Br-sil1 gene expression is induced by BTH, but not by methyl jasmonate or ethylene. This indicates that the cabbage gene is activated via a salicylic acid-dependent signaling pathway. An examination of the tissue-specific expression revealed that the induction of the Br-sil1 gene expression by BTH occurs in leaves and stems, but not in roots and flowers. Without the BTH treatment, however, the Br-sil1 gene is not expressed in any of the tissues that were examined. PMID- 14536026 TI - Molecular cloning and characterization of an NADPH quinone oxidoreductase from Kluyveromyces marxianus. AB - NAD(P)H quinone oxidoreductase is a ubiquitous enzyme that is known to directly reduce quinone substrates to hydroquinones by a two-electron reaction. We report the identification of NADPH quinone oxidoreductase from Kluyveromyces marxianus (KmQOR), which reduces quinone substrates directly to hydroquinones. The KmQOR gene was sequenced, expressed in Escherichia coli, purified, and characterized. The open-reading frame of the KmQOR gene consists of 1143 nucleotides, encoding a 380 amino acid polypeptide. The nucleotide sequence of the KmQOR gene was assigned to EMBL under accession number AY040868. The M(r) that was determined by SDS-PAGE for the protein subunit was about 42 kDa, and the molecular mass of the native KmQOR was 84 kDa, as determined by column calibration, indicating that the native protein is a homodimer. The KmQOR protein efficiently reduced 1,4 benzoquinone, whereas no activities were found for menadiones and methoxyquinones. These observations, and the result of an extended sequence analysis of known NADPH quinone oxidoreductase, suggest that KmQOR possesses a different action mechanism. PMID- 14536028 TI - Characterization of two forms of acetolactate synthase from barley. AB - Acetolactate synthase (ALS) catalyzes the first common step in the biosynthesis of valine, leucine, and isoleucine. ALS is the target site for several classes of herbicides, including sulfonylureas, imidazolinones, and triazolopyrimidines. Two forms of ALS (designated ALS I and ALS II) were separated from barley shoots by heparin affinity column chromatography. The molecular masses of native ALS I and ALS II were determined to be 248 kDa and 238 kDa by nondenaturing gel electrophoresis and activity staining. Similar molecular masses of two forms of ALS were confirmed by a Western blot analysis. SDS-PAGE and Western blot analysis showed that the molecular masses of the ALS I and ALS II subunits were identical- 65 kDa. The two ALS forms exhibited different properties with respect to the values of K(m), pI and optimum pH, and sensitivity to inhibition by herbicides sulfonylurea and imidazolinone as well as to the feedback regulation by the end product amino acids Val, Leu, and Ile. These results, therefore, suggest that the two ALS forms are not different polymeric forms of the same enzyme, but isozymes. PMID- 14536027 TI - Isolation and identification of an antioxidant enzyme catalase stimulatory compound from Garnoderma lucidum. AB - Antioxidant enzymes are scavenger reactive-oxygen intermediates and are involved in many cellular defense systems. We previously reported that a crude extract of Garnoderma lucidum, a medicinally potent mushroom, profoundly increased the catalase gene expression and enzyme activities in mouse livers (Park et al., J. Biochem. Mol. Biol. 34. 144-149, 2001). In this study, we elucidated the detailed mechanism whereby G. lucidum stimulates the catalase activity and expression. The major active fraction was isolated from G. lucidum and methyl linoleate was considered the most major component of the fraction. In order to determine whether methyl linoleate increases mRNA and protein synthesis of catalase, Northern and Western blot analyses were performed in vivo with methyl linoleate treated mouse liver homogenate after feeding methyl linoleate to the mice. Northern and Western blot analyses of the crude liver homogenates in the mice that were administered methyl linoleate revealed that the expression catalase was significantly increased when compared to the untreated controls. In addition, the catalase protein levels and enzymatic activities increased in the mouse liver homogenates. These results suggest that methyl linoleate that is produced by G. lucidum stimulates the catalase expression at the transcription level. PMID- 14536029 TI - Use of RAPD fingerprinting for discriminating two populations of Hilsa shad (Tenualosa ilisha Ham.) from inland rivers of Bangladesh. AB - The Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) was applied to analyze the genetic variation of the Hilsa shad, Tenualosa ilisha Ham., from the two major inland rivers (Padma and Meghna) in Bangladesh. Twenty eight random 10-mer primers were primarily scored in 8 individuals from each of the two locations. Fifteen primers, which gave polymorphism, were selected and used in the final analysis of 34 individuals from the two sites. Using these primers, 480 scorable DNA fragments were found, of which 98 (20.41%) were polymorphic. By comparing the RAPD banding patterns, variations were found between and within the populations. A dendrogram was constructed with the polymorphic fragments to analyze the genetic distances between the Hilsa shad populations. The results show two major clusters of Padma and Meghna, assuming different spawning populations with different stocks or races of Hilsa shad in the major Bangladesh rivers. PMID- 14536030 TI - Cobalt chloride-induced apoptosis and extracellular signal-regulated protein kinase activation in human cervical cancer HeLa cells. AB - The molecular mechanism of hypoxia-induced apoptosis has not been clearly elucidated. In this study, we investigated the involvement of extracellular signal-regulated protein kinase (ERK 1/2) in hypoxia-induced apoptosis using cobalt chloride in HeLa human cervical cancer cells. The cobalt chloride was used for the induction of hypoxia, and its IC(50) was 471.4 microM. We demonstrated the DNA fragmentation after incubation with concentrations more than 50 microM cobalt chloride for 24 h, and also evidenced the morphological changes of the cells undergoing apoptosis with electron microscopy. Next, we examined the signaling pathway of cobalt chloride-induced apoptosis in HeLa cells. ERK1/2 activation occurred 6 and 9 h after treatment with 600 microM cobalt chloride. Meanwhile, the pretreatment of the MEK 1 inhibitor (PD98059) completely blocked the cobalt chloride-induced ERK 1/2 activation. At the same time, the activated ERK 1/2 translocated into the nucleus and phosphorylated its transcriptional factor, c-Jun. In addition, the pretreatment of PD98059 inhibited the cobalt chloride-induced DNA fragmentation and apoptotic cell death. These results suggest that cobalt chloride is able to induce apoptotic activity in HeLa cells, and its apoptotic mechanism may be associated with signal transduction via ERK 1/2. PMID- 14536031 TI - Choristoneura fumiferana Granulovirus p74 protein, a highly conserved baculoviral envelope protein. AB - A gene that encodes a homologue to baculoviral p74, an envelope-associated viral structural protein, has been identified and sequenced on the genome of Choristoneura fumiferana granulovirus (ChfuGV). A part of the ChfuGV p74 gene was located on an 8.9 kb BamHI subgenomic fragment using different sets of degenerated primers. These were designed using the results of the protein sequencing of a major 74 kDa structural protein that is associated with the occlusion-derived virus (ODV). The gene has a 1992 nucleotide (nt) open-reading frame (ORF) that encodes a protein with 663 amino acids with a predicted molecular mass of 74,812 Da. Comparative studies revealed the presence of two major conserved regions in the ChfuGV p74 protein. This study also shows that all of the p74 proteins contain two putative transmembrane domains at their C terminal segments. At the nucleotide sequence level, two late promoter motifs (TAAG and GTAAG) were located upstream of the first ATG of the p74 gene. The gene contained a canonical poly(A) signal, AATAAA, at its 3 non-translated region. A phylogenetic tree for baculoviral p74 was constructed using a maximum parsimony analysis. The phylogenetic estimation demonstrated that ChfuGV p74 is related the closest to those of Cydia pomonella granulovirus (CpGV) and Phthorimaea operculella granulovirus (PhopGV). PMID- 14536032 TI - Oxidative modification of neurofilament-L by copper-catalyzed reaction. AB - Neurofilament-L (NF-L) is a major element of neuronal cytoskeletons and known to be important for neuronal survival in vivo. Since oxidative stress might play a critical role in the pathogenesis of neurodegenerative diseases, we investigated the role of copper and peroxide in the modification of NF-L. When disassembled NF L was incubated with copper ion and hydrogen peroxide, then the aggregation of protein was proportional to copper and hydrogen peroxide concentrations. Dityrosine crosslink formation was obtained in copper-mediated NF-L aggregates. The copper-mediated modification of NF-L was significantly inhibited by thiol antioxidants, Nacetylcysteine, glutathione, and thiourea. A thioflavin-T binding assay was performed to determine whether the copper/H2O2 system-induced in vitro aggregation of NF-L displays amyloid-like characteristics. The aggregate of NF-L displayed thioflavin T reactivity, which was reminiscent of amyloid. This study suggests that copper-mediated NF-L modification might be closely related to oxidative reactions which may play a critical role in neurodegenerative diseases. PMID- 14536033 TI - Bacterial expression of the scFv fragment of a recombinant antibody specific for Burkholderia pseudomallei exotoxin. AB - The scFv antibody towards the Burkholderia pseudomallei exotoxin was previously constructed by phage display and exhibited good specificity towards the exotoxin. We report here the optimization of the scFv expression in an E. coli expression system. Four different E. coli strains (ER2537, TG1, HB2151, and XL1-Blue) were examined for optimal expression of the scFv protein. Two types of carbon source (i.e. 0.2% glucose and 0.2% glycerol) were also tested for their ability to induce the scFv expression. Cells that carried the scFv construct were grown at 30 degrees C and induced with 0.05 mM IPTG. The expression was then monitored by SDS-PAGE, Western blotting, and indirect ELISA. The Western blot profile showed different levels of the scFv expression among the host strains; XL1-Blue exhibited the highest level of the scFv protein expression. Glycerol at a concentration of 0.2% (v/v) significantly increased the scFv protein expression level when compared to 0.2% (w/v) glucose. Further optimization demonstrated that the scFv protein expression in XL1-Blue was the most optimal with a glycerol concentration as low as 0.05%. However, by indirect ELISA, only the scFv protein that was expressed in 0.2% (v/v) glycerol exhibited high specificity towards the Burkholderia pseudomallei exotoxin. PMID- 14536034 TI - Effect of diazinon, an organophosphate insecticide, on plasma lipid constituents in experimental animals. AB - There has been increasing interest in studying the various effects of organophosphate insecticides in humans and experimental animals. Only a few data are available on the effect of the organophosphate insecticide, diazinon, on lipid metabolism. The aim of this study was to evaluate the effect of diazinon on plasma lipid constituents in mammalian animals. The plasma levels of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), triglycerides (TG), and phospholipids (PL) were measured in albino rats that were orally treated with a single dose of diazinon at a level of LD(50) or with repeated daily doses at the levels of 1/2, 1/8, and 1/32 LD(50) for 2, 8, and 32 days, respectively. After a 24 h post-treatment with a single LD(50) dose of diazinon, TC was not significantly changed, the HDL-C and PL levels were significantly decreased, but the LDL-C and TG levels were significantly increased. Separate daily oral administrations of diazinon at 1/2 LD(50), 1/8 LD(50), and 1/32 LD(50) doses resulted in a significant decrease in HDL-C and PL, with no significant change in TG. The LDL-C levels were significantly increased and TC showed no significant change with 1/2 LD(50) and 1/32 LD(50) doses of diazinon, whereas a significant decrease in the levels of TC, HDL-C, as well as LDL-C, was observed with the 1/8 LD(50) dose. These data suggest that diazinon may interfere with lipid metabolism in mammals. PMID- 14536035 TI - Backbone 1H, 15N, and 13C resonance assignments of the Helicobacter pylori acyl carrier protein. AB - One of the small proteins from Helicobacter pylori, acyl carrier protein (ACP), was investigated by NMR. ACP is related to various cellular processes, especially with the biosynthesis of fatty acid. The basic NMR resonance assignment is a prerequisite for the validation of a heterologous protein interaction with ACP in H. pylori. Here, the results of the backbone (1)H, (15)N, and (13)C resonance assignments of the H. pylori ACP are reported using double- and triple-resonance techniques. About 97% of all of the (1)HN, (15)N, (13)CO, (13)Calpha, and (13)Cbeta resonances that cover 76 of the 78 non-proline residues are clarified through sequential- and specific- assignments. In addition, four helical regions were clearly identified on the basis of the resonance assignments. PMID- 14536036 TI - Identification of two isoforms of aminopeptidase N in Aedes aegypti larval midgut. AB - The bacterium Bacillus thuringiensis produces toxin inclusions that are deleterious to target insect larvae. These toxins are believed to interact with a specific receptor protein(s) that is present on the gut epithelial cells of the larvae. In various insect species (in particular those belonging to the lepidopteran class), aminopeptidase N (APN) is one of the two receptor proteins that are considered to be involved in toxin-receptor interactions. However, in mosquitoes, the nature and identity of the receptor protein is unknown. Here, using RT-PCR, we identified two isoforms of the APN transcripts in the Aedes aegypti mosquito larval midgut. These results are congruent with a previous report of multiple isoforms of the APN gene expression in lepidopteran larvae. Which of the two isoforms (or other yet unidentified receptor proteins) is involved in the killing of mosquito larvae remains to be elucidated. PMID- 14536037 TI - Cloning of a ribonucleotide reductase gene of the herpes simplex virus type 2 strain G. AB - The ribonucleotide reductase (RR) 2 gene of the HSV-2 strain G was cloned, sequenced, and expressed in an E. coli cell. The RR2 gene was located on the PstI 2.4 kb fragment, which was cloned and sequenced. The ORF of the gene was 1,011 bp and its termination codon was TAG; also, the CATATAA sequence was present in the promoter of the RR2 gene. A Poly A signal sequence (AATAAA) was found in the 3' noncoding region. The RR2 proteins that were produced in the E. coli and Vero cells were confirmed using a Western blot analysis. SDS-PAGE revealed that the molecular weights of the fusion-RR2 that was produced in the E. coli cells were approximately 24 kDa and 38 kDa in the Vero cells. The RR2 proteins were soluble. The differences in the molecular weights might be due to modifications in the Vero cells. PMID- 14536038 TI - Production of nuclease activity in U937 cells by phorbol 12-myristate 13-acetate and lipopolysaccharide. AB - The proliferation and differentiation signals of myelogeneous U937 cells are provided by extracellular stimuli, such as lipopolysaccharide (LPS) and phorbol 12-myristate 13-acetate (PMA). In a DNA-native-polyacrylamide gel assay system, we demonstrated that a particular nuclease activity is expressed in PMA stimulated U937 cells and secreted into the culture medium. The nuclease activity was induced in U937 cells by LPS treatment, while the secretion of the enzyme was undetected in the culture medium. Therefore, it is likely that the expression and secretion of the particular nuclease in U937 cells are controlled by extracellular stimulations, such as PMA and LPS treatment. PMID- 14536039 TI - Measurements of buccal tissue volumes at single-implant restorations after local bone grafting in maxillas: a 3-year clinical prospective study case series. AB - PURPOSE: The purpose of this study was to measure changes in buccal and proximal tissue volumes after local bone grafting and single-implant treatment. MATERIALS AND METHODS: Ten patients were provided with buccal bone grafts 6 months prior to implant treatment in central upper incisor regions. Following a healing time of 6 months, abutments and single-implant crowns were installed and followed up for 2 years. Clinical photographs and impressions were taken prior to the surgical intervention as well as after crown placement and at first and second annual checkups. The photographs and study models were analyzed with regard to papilla regeneration and changes in buccal crest volume during the study period by means of a clinical papilla index and optical scanning of study models. RESULTS: All bone grafts healed without problems. A significant reduction of the buccal crest volume (-50%, p <.01) was observed in the grafted area before abutment connection. However, a significant increase of tissue volume (+100%, p <.05) was noticed at the subsequent crown placement, followed by a second but slow reduction of the volume during the following 2 years of function. The interdental papillae increased significantly (p <.05) in volume during the first year, almost completely filling up the embrasure areas after 2 years. CONCLUSIONS: It may be concluded that local bone grafting seems to be a valuable protocol to create sufficient bone volume for implant placement. However, significant resorption of the graft may be present, which reduces the impact of grafting on the esthetic outcome. Instead, placement of the abutment cylinder and the crown seems to play a more important role for reestablishing the tissue volume at the implant supported single crowns. PMID- 14536040 TI - One-year prospective three-center study comparing the outcome of a "soft bone implant" (prototype Mk IV) and the standard Branemark implant. AB - BACKGROUND: Oral implant treatment ad modum Branemark has been used for decades in the rehabilitation of edentate and partially dentate patients. Posterior jaw regions frequently exhibit bone of poor texture, and it is often difficult to obtain primary stability. Thus, it may prove beneficial to deviate from the original protocol and to use implants with a modified design, for example, with a slightly tapered geometry. PURPOSE: The purpose of the investigation was to compare the early behavior of a modified (prototype Mk IV, Branemark System, Nobel Biocare AB, Gothenburg, Sweden; test) implant with that of the standard Branemark implant (control) in regions of mainly type 4 bone. MATERIALS AND METHODS: Three Swedish centers participated, and a total of 44 patients were treated with oral implants for 39 maxillas and 5 mandibles. The study focused on the most distal right and left implant sites (88 implants), which were randomized to receive either a test or a control implant. Various parameters were recorded, such as registered insertion torque (OsseoCare), Nobel Biocare AB), wobbling during insertion, primary and secondary stability (as measured with resonance frequency), and marginal bone loss. The implants were followed up for 1 year. RESULTS: The test implant more frequently required a higher insertion torque and showed a significantly higher primary stability than the control implant. This difference in stability leveled out over time, and test and control implants exhibited similar secondary stability at abutment operation and at the 1-year visit. Wobbling during insertion was rarely recorded for either of the implant designs. The 1-year cumulative success rate was 93.1% for test implants and 88.4% for control implants. CONCLUSIONS: The modified implant design resulted in an increased primary stability, which may be important when placing implants in jaw regions of type 4 bone. However, independent of the achieved primary stability, successful implants tended to approach similar secondary stability in the two designs tested. PMID- 14536041 TI - Spontaneous bone formation in the maxillary sinus after removal of a cyst: coincidence or consequence? AB - BACKGROUND: Maxillary sinus floor-augmentation techniques are frequently used to increase the bone volume in the posterior edentulous maxilla to enable placement and integration of titanium implants. PURPOSE: The purpose of this report is to document an unexpected healing pattern after maxillary sinus surgery and to discuss the implications for future bone-augmentation techniques. MATERIALS AND METHODS: In a patient referred for sinus augmentation, an intrasinus mucosal cyst was removed 3 months prior to the planned augmentation procedure. A replaceable bone window was prepared in the lateral aspect of the sinus wall. The cyst was removed, the ruptured mucosa was sutured, and the bone window was replaced, resulting in a secluded space in the sinus. RESULTS: After 3 months of healing, the space between the replaced bony window and the lifted sinus membrane was filled with newly formed bone. The surgical technique was repeated in a second patient and resulted in a similar bone reformation pattern. CONCLUSION: Surgical trauma and the creation of a secluded space between the bone surfaces and the sinus mucosa result in spontaneous bone formation in the maxillary sinus. The surgical approach described may be used to achieve bone reformation to enable placement of dental implants without the addition of any grafting material. PMID- 14536042 TI - Tissue reactions to abutment shift: an experimental study in dogs. AB - BACKGROUND: Standard protocols for the clinical use of dental implants often include the placement of healing abutments prior to standard or custom-made abutments. The tissue response to a single shift from a healing abutment to a permanent abutment has not been studied. PURPOSE: The aim of the present experiment was to study tissue reactions that may occur following the removal of a healing abutment and the placement of a permanent abutment. MATERIALS AND METHODS: In six beagle dogs, all mandibular premolars were extracted. Three months later three fixtures of the Astra Tech Implants Dental System (Astra Tech AB, Molndal, Sweden) were installed in each edentulous premolar region. An additional 3 months later, the first abutment connection was performed. In two sites on each side of the mandible, healing abutments were placed; in the remaining site, a Uni-abutment (Astra Tech AB) was used. The two healing abutments were removed 2 weeks later, and one Uni-abutment and one prepable abutment were placed. A plaque-control period was initiated, and 6 months later block biopsies were obtained. The biopsies were prepared for histometric and morphometric examination. Radiographs were obtained at fixture placement, 2 weeks after the first abutment connection, and 6 months later. RESULTS: The length of the barrier epithelium, the height of the connective tissue attachment, and the level of the marginal bone did not differ between the three abutment groups. The major part of the radiographic bone loss during the experiment took place prior to or immediately after abutment connection; only small bone level alterations occurred during the subsequent 6-month period. CONCLUSIONS: The shift from a healing abutment to a permanent abutment resulted in the establishment of a transmucosal attachment, the dimension and quality of which did not differ from those of the mucosal barrier formed to a permanent abutment placed during a second-stage surgery. PMID- 14536043 TI - Histologic analysis of clinically retrieved immediately loaded titanium implants: a report of 11 cases. AB - BACKGROUND: Several investigators have reported high survival rates for early or immediately loaded dental implants. PURPOSE: The aim of the present study was to analyze histologically the periimplant tissue reactions and the bone-titanium interface of clinically retrieved immediately loaded titanium implants. MATERIALS AND METHODS: Eleven implants were inserted in the posterior jaw regions of six patients, serving as the most distal abutments of fixed provisional bridges and subjected to immediate occlusal loading. After a 10-month loading period, all 11 implants were retrieved with a trephine. RESULTS: Mature bone was present at the interface of all implants. Some vertical bone loss was observed; this was more pronounced for cylindric implants. The bone-implant contact was approximately 60 to 65% for all implants. CONCLUSIONS: The results demonstrate that osseointegration of dental implants can occur during immediate loading. PMID- 14536044 TI - Resorbable and nonresorbable hydroxyapatite granules as bone graft substitutes in rabbit cortical defects. AB - BACKGROUND: The use of various synthetic calcium phosphate compositions for the promotion of bone in bone defects is of potential interest because such materials may be tailor made and may bond to bone. There is yet an inadequate knowledge of the role of calcium phosphate composition and resorbability for the bone response. PURPOSE: The aim of the present study was to compare the ability of resorbable versus nonresorbable hydroxyapatite (HA) granules to promote new bone formation in cortical bone defects. Resorbable and nonresorbable HA granules, used as bone graft substitutes, were evaluated after 6 weeks and 3 months in the rabbit tibia. Circular defects (diameter 5.0 mm) were made in both tibias of 18 New Zealand white rabbits. The 36 defects were divided into three groups (six observations per group and time, respectively). The first group was augmented with resorbable HA granules, the second group was augmented with ceramic nonresorbable HA granules, and the third group was left without augmentation (control). The animals were killed after 6 weeks and 3 months, and the tissue was evaluated with light microscopic (LM) morphology and morphometry, scanning electron microscopy (SEM), and energy dispersive x-ray analysis (EDX). RESULTS: After 3 months LM morphometry revealed significantly more newly formed bone in the two HA augmented groups compared with that in the control. A close contact was found between both kinds of HA granules and new bone as viewed with light microscopy and SEM. A relatively slow degradation process was indicated by the small reduction of the total granule area in the cortical defects. However, LM observations showed a change of granule form. Pilot experiments using SEM-EDX indicate that Ca and P contents had decreased in the resorbable HA granules between 6 weeks and 3 months. Further, a higher content of Ca and P was found in the newly formed bone close to granules, in comparison with more distant newly formed bone. CONCLUSIONS: Our results suggest that both resorbable and nonresorbable HA granules promote new bone formation in rabbit cortical defects, which does not occur in control defects. PMID- 14536045 TI - Bone healing around titanium and titanium nitride-coated dental implants with three surfaces: an experimental study in rats. AB - BACKGROUND: Titanium nitride (TiN) has been used in many fields as a coating of surgical instruments, with the purpose of creating materials more resistant to wear and corrosion and also reducing adhesion. PURPOSE: The aim of this study was to evaluate the biocompatibility of TiN-coated dental implants. MATERIALS AND METHODS: Forty-five rats were used in this study. One hundred eighty 2 mm x 2 mm implants (P.H.I. San Vittore Olona, Milano, Italy) were used. The implants were divided into the following three groups: Group 1 (n = 60): 30 machined and 30 machined coated with Group 2 (n = 60): 30 sandblasted and 30 sandblasted coated with Group 3 (n = 60): 30 titanium plasma sprayed, 30 titanium plasma sprayed and coated with TiN Four implants were placed in each rat, two implants coated with TiN on the right tibia and two uncoated implants on the left. The animals were killed after 5, 10, 20, 30, or 60 days. Another 18 implants were used for surface roughness analysis. RESULTS: The present study showed that the healing around the TiN-coated implants was similar to that observed around the uncoated surfaces. CONCLUSIONS: TiN coating demonstrated a good biocompatibility, did not have untoward effects on the periimplant bone formation, and did not change the surface roughness values. PMID- 14536046 TI - Space-providing expanded polytetrafluoroethylene devices define alveolar augmentation at dental implants induced by recombinant human bone morphogenetic protein 2 in an absorbable collagen sponge carrier. AB - BACKGROUND: Surgical implantation of recombinant human bone morphogenetic protein 2 (rhBMP-2) in an absorbable collagen sponge carrier (ACS) significantly enhances bone regeneration in horizontal alveolar defects; however, sufficient quantities of bone for implant dentistry are not routinely obtained. PURPOSE: The objective of this proof-of-principle study was to evaluate the potential of a space providing macroporous expanded polytetrafluoroethylene (ePTFE) device to control volume and geometry of rhBMP-2/ACS-induced alveolar bone augmentation. MATERIALS AND METHODS: Bilateral critical-size supra-alveolar periimplant defects were created in four Hound-Labrador mongrel dogs. Two turned and one surface-etched 10 mm titanium dental implants were placed 5 mm into the surgically reduced alveolar ridge creating 5 mm supra-alveolar defects. rhBMP-2/ACS (0.4 mg rhBMP-2) was placed around the exposed dental implants. Additionally, one jaw quadrant in each animal was randomly assigned to receive the dome-shaped macroporous ePTFE device. Mucoperiosteal flaps were advanced for primary wound closure. The animals were euthanized at 8 weeks post surgery for histometric analysis. RESULTS: The space providing macroporous ePTFE device defined the volume and geometry of rhBMP-2/ACS induced bone formation, whereas bone formation at sites receiving rhBMP-2/ACS alone varied considerably. Vertical bone gain at turned dental implants averaged (+/-SD) 4.7 +/-0.2 mm at sites receiving rhBMP-2/ACS and the ePTFE device compared with 3.5 +/-0.9 mm at sites receiving rhBMP-2/ACS only. The corresponding values for rhBMP-2/ACS-induced bone area were 9.6 +/- 0.7 mm2 and 7.5 +/-6.2 mm2. There was a highly significant correlation between induced bone area and the space provided by the ePTFE device (p <.001). There was no difference in induced bone density or bone-implant contact between the two technologies. These observations were consistent with those observed at surface etched dental implants. CONCLUSIONS: The data from this study suggest that a space-providing macroporous ePTFE device defines rhBMP-2/ACS-induced alveolar augmentation to provide adequate bone quantities for implant dentistry. The dental implant surface technology does not appear to substantially influence bone formation. PMID- 14536047 TI - Periimplant bone healing under experimental hepatic osteodystrophy induced by a choline-deficient diet: a histomorphometric study in rats. AB - BACKGROUND: Osseointegration and success of an implant involve the interaction of local and systemic factors such as bone metabolic diseases. PURPOSE: The purpose of this study was to evaluate the effect of experimental hepatic osteodystrophy induced by a choline-deficient diet on periimplant bone healing. MATERIALS AND METHODS: Laminar titanium implants were placed in the tibias of five groups of Wistar rats: those with a (1). controlled diet for 15 days; (2). choline deficient diet for 15 days; (3). controlled diet for 30 days; (4). choline deficient diet for 30 days; (5). choline-deficient diet for 15 days and a controlled diet for 15 days (refeeding). Body weight and food intake, hematocrit, and hemoglobinemia were evaluated. The animals were killed at 15 or 30 days post implantation. The liver, kidneys, and tibias were resected and fixed in 20% formalin solution. The tibias were radiographed and processed for histomorphometric evaluation of the periimplant bone area. RESULTS: Histologic studies revealed steatosis in the liver but no alterations in the kidneys. Rats fed a choline-deficient diet showed periimplant bone healing with marked qualitative and quantitative alterations. The periimplant bone area was 28% and 75% lower in experimental animals than in controls at 15 and 30 days, respectively. CONCLUSIONS: Liver alterations caused by a choline-deficient diet alter periimplant osteogenesis qualitatively and quantitatively. PMID- 14536048 TI - Immediate replacement of single upper posterior teeth: a report of cases. AB - BACKGROUND: The immediate replacement of upper molar teeth with implants poses a significant challenge to the clinician because of its sinus involvement. PURPOSE: The purpose of this article is to report a protocol in which single molar teeth are replaced immediately with implants, minimizing the need for other surgical procedures. MATERIALS AND METHODS: This protocol has been employed in eight cases. Briefly, the teeth are extracted atraumatically, the sockets evaluated, an osteotomy performed following the direction of the socket, and a wide diameter implant placed using the socket walls and sinus floor to anchor it. A grafting material is placed when the space between implant and socket wall is considerably large. Three cases are reviewed here, and all eight are summarized. RESULTS: All cases have excellent soft and hard tissue healing after 6 months. Furthermore, the overall time needed to restore the cases has been considerably reduced and the need for sinus management eliminated. CONCLUSION: The cases presented support the use of the described protocol in the placement of implants in maxillary molar areas without any other sinus procedures. PMID- 14536050 TI - Myoblasts fuse when loner meets ARF6. AB - Studies of myoblast fusion in Drosophila have revealed numerous signaling and structural proteins that regulate this critical event in muscle development. A paper by Chen et al. published in the September 19 issue of Cell adds ARF6, a GTPase, and one of its guanine nucleotide exchange factors to this set of players. PMID- 14536049 TI - Hand-me-downs. Transmitting anterior-posterior asymmetry from older to younger egg follicles. AB - In Drosophila, the initial asymmetries that determine the embryonic body axes are generated during oogenesis. A report in this issue of Developmental Cell shows that a developing follicle conveys its anterior-posterior asymmetry to the next younger follicle via a relay mechanism that involves the Notch/Delta and the JAK/STAT signaling pathways. PMID- 14536051 TI - Ac'septin' a signal: kinase regulation by septins. AB - Budding yeast monitor shape and the assembly of cytoskeletal structures and convey this information to regulators of cell division, but the molecular mechanisms responsible for monitoring and interpreting spatial information about the cytoskeleton remain poorly understood. A paper in the September issue of Molecular Cell shows that direct binding of components of the septin cytoskeleton may relieve autoinhibition of a conserved checkpoint kinase, creating a simple molecular device for sensing septin cytoskeleton organization. PMID- 14536052 TI - AP-2 makes room for rivals. AB - Clathrin and the adaptor protein complex (AP-2) constitute the major coat components of clathrin-coated vesicles. In the September issues of the Journal of Cell Biology and the Journal of Biological Chemistry, three reports reveal that AP-2, while essential for internalization of transferrin, is not essential for internalization of EGF. These novel data suggest the intriguing possibility that the major role of AP-2 is in cargo recruitment, and not in assembly of functionally active clathrin-coated pits. PMID- 14536053 TI - Stem cells in the news: CNN and APC make headlines. AB - Stem cell self-renewal depends on their ability to divide asymmetrically, with one daughter retaining stem cell identity. This often involves precise orchestration of mitotic spindle orientation, but the machinery used to ensure this outcome is understood in only a handful of examples. In a recent issue of Science, Yamashita et al. provide new insights into the factors that control this process in the male germline of Drosophila, identifying roles for the centrosomal protein Centrosomin and tumor suppressor homologs of the APC family. PMID- 14536054 TI - RNAi on the apoptosis TRAIL: the mammalian cell genetic screen comes of age. AB - Presented in a paper in the September issue of Molecular Cell, Aza-Blanc et al. used an RNA interference-based genetic screen to identify genes that modulate sensitivity to the apoptosis-inducing ligand TRAIL. The age of mammalian cell genetic screens has begun. PMID- 14536055 TI - Ephective endocytosis. AB - Two important new reports identify endocytosis of EphB-ephrinB complexes as a mechanism for switching between cell-cell adhesion and repulsion following plasma membrane contact. Together with the previously described shedding of ephrinA following EphA engagement, these findings resolve the paradox of how an adhesive receptor-ligand interaction generates a repulsive cellular response. PMID- 14536056 TI - A unified nomenclature for yeast autophagy-related genes. PMID- 14536057 TI - A Notch/Delta-dependent relay mechanism establishes anterior-posterior polarity in Drosophila. AB - The anterior-posterior axis of Drosophila becomes polarized early in oogenesis, when the oocyte moves to the posterior of the germline cyst because it preferentially adheres to posterior follicle cells. The source of this asymmetry is unclear, however, since anterior and posterior follicle cells are equivalent until midoogenesis, when Gurken signaling from the oocyte induces posterior fate. Here, we show that asymmetry arises because each cyst polarizes the next cyst through a series of posterior to anterior inductions. Delta signaling from the older cyst induces the anterior polar follicle cells, the anterior polar cells signal through the JAK/STAT pathway to induce the formation of the stalk between adjacent cysts, and the stalk polarizes the younger anterior cyst by inducing the shape change and preferential adhesion that position the oocyte at the posterior. The anterior-posterior axis is therefore established by a relay mechanism, which propagates polarity from one cyst to the next. PMID- 14536058 TI - Phospholipid membrane composition affects EGF receptor and Notch signaling through effects on endocytosis during Drosophila development. AB - The role of phospholipids in the regulation of membrane trafficking and signaling is largely unknown. Phosphatidylcholine (PC) is a main component of the plasma membrane. Mutants in the Drosophila phosphocholine cytidylyltransferase 1 (CCT1), the rate-limiting enzyme in PC biosynthesis, show an altered phospholipid composition with reduced PC and increased phosphatidylinositol (PI) levels. Phenotypic features of dCCT1 indicate that the enzyme is not required for cell survival, but serves a role in endocytic regulation. CCT1- cells show an increase in endocytosis and enlarged endosomal compartments, whereas lysosomal delivery is unchanged. As a consequence, an increase in endocytic localization of EGF receptor (Egfr) and Notch is observed, and this correlates with a reduction in signaling strength and leads to patterning defects. A further link between PC/PI content, endocytosis, and signaling is supported by genetic interactions of dCCT1 with Egfr, Notch, and genes affecting endosomal traffic. PMID- 14536059 TI - Rab7 prevents growth factor-independent survival by inhibiting cell-autonomous nutrient transporter expression. AB - Growth factor withdrawal results in the endocytosis and degradation of transporter proteins for glucose and amino acids. Here, we show that this process is under the active control of the small GTPase Rab7. In the presence of growth factor, Rab7 inhibition had no effect on nutrient transporter expression. In growth factor-deprived cells, however, blocking Rab7 function prevented the clearance of glucose and amino acid transporter proteins from the cell surface. When Rab7 was inhibited, growth factor deprived cells maintained their mitochondrial membrane potential and displayed prolonged, growth factor independent, nutrient-dependent cell survival. Thus, Rab7 functions as a proapoptotic protein by limiting cell-autonomous nutrient uptake. Consistent with this, dominant-negative Rab7 cooperated with E1A to promote the transformation of p53(-/-) mouse embryonic fibroblasts (MEFs). These results suggest that proteins that limit nutrient transporter expression function to prevent cell-autonomous growth and survival. PMID- 14536060 TI - ER-to-Golgi carriers arise through direct en bloc protrusion and multistage maturation of specialized ER exit domains. AB - Protein transport between the ER and the Golgi in mammalian cells occurs via large pleiomorphic carriers, and most current models suggest that these are formed by the fusion of small ER-derived COPII vesicles. We have examined the dynamics and structural features of these carriers during and after their formation from the ER by correlative video/light electron microscopy and tomography. We found that saccular carriers containing either the large supramolecular cargo procollagen or the small diffusible cargo protein VSVG arise through cargo concentration and direct en bloc protrusion of specialized ER domains in the vicinity of COPII-coated exit sites. This formation process is COPII dependent but does not involve budding and fusion of COPII-dependent vesicles. Fully protruded saccules then move centripetally, evolving into one of two types of carriers (with distinct kinetic and structural features). These findings provide an alternative framework for analysis of ER-to-Golgi traffic. PMID- 14536061 TI - Differential roles of WAVE1 and WAVE2 in dorsal and peripheral ruffle formation for fibroblast cell migration. AB - Cell migration is driven by actin polymerization at the leading edge of lamellipodia, where WASP family verprolin-homologous proteins (WAVEs) activate Arp2/3 complex. When fibroblasts are stimulated with PDGF, formation of peripheral ruffles precedes that of dorsal ruffles in lamellipodia. Here, we show that WAVE2 deficiency impairs peripheral ruffle formation and WAVE1 deficiency impairs dorsal ruffle formation. During directed cell migration in the absence of extracellular matrix (ECM), cells migrate with peripheral ruffles at the leading edge and WAVE2, but not WAVE1, is essential. In contrast, both WAVE1 and WAVE2 are essential for invading migration into ECM, suggesting that the leading edge in ECM has characteristics of both ruffles. WAVE1 is colocalized with ECM degrading enzyme MMP-2 in dorsal ruffles, and WAVE1-, but not WAVE2-, dependent migration requires MMP activity. Thus, WAVE2 is essential for leading edge extension for directed migration in general and WAVE1 is essential in MMP dependent migration in ECM. PMID- 14536062 TI - The claudin-like megatrachea is essential in septate junctions for the epithelial barrier function in Drosophila. AB - Vertebrate claudin proteins are integral components of tight junctions, which function as paracellular diffusion barriers in epithelia. We identified Megatrachea (Mega), a Drosophila transmembrane protein homologous to claudins, and show that it acts in septate junctions, the corresponding structure of invertebrates. Our analysis revealed that Mega has transepithelial barrier function similar to the claudins. Also, Mega is necessary for normal tracheal cell morphogenesis but not for apicobasal polarity or epithelial integrity. In addition, we present evidence that Mega is essential for localization of the septate junction protein complex Coracle/Neurexin. The results indicate that claudin-like proteins are functionally conserved between vertebrates and Drosophila. PMID- 14536063 TI - Otx/otd homeobox genes specify distinct sensory neuron identities in C. elegans. AB - The mechanisms by which the diverse functional identities of neurons are generated are poorly understood. C. elegans responds to thermal and chemical stimuli using 12 types of sensory neurons. The Otx/otd homolog ttx-1 specifies the identities of the AFD thermosensory neurons. We show here that ceh-36 and ceh 37, the remaining two Otx-like genes in the C. elegans genome, specify the identities of AWC, ASE, and AWB chemosensory neurons, defining a role for this gene family in sensory neuron specification. All C. elegans Otx genes and rat Otx1 can substitute for ceh-37 and ceh-36, but only ceh-37 functionally substitutes for ttx-1. Functional substitution in the AWB neurons is mediated by activation of the same downstream target lim-4 by different Otx genes. Misexpression experiments indicate that although the specific identity adopted upon expression of an Otx gene may be constrained by the cellular context, individual Otx genes preferentially promote distinct neuronal identities. PMID- 14536064 TI - Self-enhanced ligand degradation underlies robustness of morphogen gradients. AB - Morphogen gradients provide long-range positional information by extending across a developing field. To ensure reproducible patterning, their profile is invariable despite genetic or environmental fluctuations. Common models assume a morphogen profile that decays exponentially. Here, we show that exponential profiles cannot, at the same time, buffer fluctuations in morphogen production rate and define long-range gradients. To comply with both requirements, morphogens should decay rapidly close to their source but at a significantly slower rate over most of the field. Numerical search revealed two network designs that support robustness to fluctuations in morphogen production rate. In both cases, morphogens enhance their own degradation, leading to a higher degradation rate close to their source. This is achieved through reciprocal interactions between the morphogen and its receptor. The two robust networks are consistent with properties of the Wg and Hh morphogens in the Drosophila wing disc and provide novel insights into their function. PMID- 14536065 TI - Quorum sensing-dependent biofilms enhance colonization in Vibrio cholerae. AB - Vibrio cholerae is the causative agent of the diarrheal disease cholera. By an incompletely understood developmental process, V. cholerae forms complex surface associated communities called biofilms. Here we show that quorum sensing deficient mutants of V. cholerae produce thicker biofilms than those formed by wild-type bacteria. Microarray analysis of biofilm-associated bacteria shows that expression of the Vibrio polysaccharide synthesis (vps) operons is enhanced in hapR mutants. CqsA, one of two known autoinducer synthases in V. cholerae, acts through HapR to repress vps gene expression. Vibrio biofilms are more acid resistant than planktonic cells. However, quorum sensing-deficient biofilms have lower colonization capacities than those of wild-type biofilms, suggesting that quorum sensing may promote cellular exit from the biofilm once the organisms have traversed the gastric acid barrier of the stomach. These results shed light on the relationships among biofilm development, quorum sensing, infectivity, and pathogenesis in V. cholerae. PMID- 14536066 TI - Genetic modulation of PPARgamma phosphorylation regulates insulin sensitivity. AB - Obesity-associated diabetes is epidemic in industrialized societies. The nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma) is highly expressed in adipose tissue and the presumed molecular target for antidiabetic thiazolidinedione drugs that reverse insulin resistance but also promote weight gain. Phosphorylation reduces the activity of PPARgamma in vitro, but physiological relevance has not been demonstrated. We have studied mice homozygous for a mutation (S112A) that prevents PPARgamma phosphorylation. Surprisingly, the weights and adipose mass of PPARgamma-S112A mice are not greater than wild-type. Remarkably, however, genetic prevention of PPARgamma phosphorylation preserves insulin sensitivity in the setting of diet-induced obesity. Underlying this protection are smaller fat cells, elevated serum adiponectin, and reduced free fatty acid levels. Thus, the phosphorylation state of PPARgamma modulates insulin sensitivity. Compounds that prevent PPARgamma phosphorylation or ligands that induce the conformation of nonphosphorylated PPARgamma may selectively enhance insulin sensitivity without increasing body weight. PMID- 14536067 TI - Integration of growth factor and nutrient signaling: implications for cancer biology. AB - Signaling networks that promote cell growth are frequently dysregulated in cancer. One regulatory network, which converges on effectors such as 4EBP1 and S6K1, leads to growth by promoting protein synthesis. Here, we discuss how this network is regulated by both extracellular signals, such as growth factors, and intracellular signals, such as nutrients. We discuss how mutations amplifying either type of signal can lead to tumor formation. In particular, we focus on the recent discovery that a tumor suppressor complex whose function is lost in tuberous sclerosis patients regulates the nutrient signal carried by the critical signaling protein TOR to the effectors 4EBP1 and S6K1. Finally, we describe how the small molecule rapamycin, which inhibits TOR and thereby the activation of these effectors, could be useful to treat tumors that have become dependent upon this pathway for growth. PMID- 14536068 TI - Chromatin and epigenetics: dynamic organization meets regulated function. AB - An EMBO workshop held June 19-22, 2003, at the European Molecular Biology Laboratory in Heidelberg, Germany, was dedicated to the memory of Alan Wolffe. It provided a venue to highlight new concepts in the establishment and maintenance of functional chromatin organization. PMID- 14536069 TI - Activation of the pyrrolysine suppressor tRNA requires formation of a ternary complex with class I and class II lysyl-tRNA synthetases. AB - Monomethylamine methyltransferase of the archaeon Methanosarcina barkeri contains a rare amino acid, pyrrolysine, encoded by the termination codon UAG. Translation of this UAG requires the aminoacylation of the corresponding amber suppressor tRNAPyl. Previous studies reported that tRNAPyl could be aminoacylated by the synthetase-like protein PylS. We now show that tRNAPyl is efficiently aminoacylated in the presence of both the class I LysRS and class II LysRS of M. barkeri, but not by either enzyme acting alone or by PylS. In vitro studies show that both the class I and II LysRS enzymes must bind tRNAPyl in order for the aminoacylation reaction to proceed. Structural modeling and selective inhibition experiments indicate that the class I and II LysRSs form a ternary complex with tRNAPyl, with the aminoacylation activity residing in the class II enzyme. PMID- 14536070 TI - A 3' exonuclease that specifically interacts with the 3' end of histone mRNA. AB - Metazoan histone mRNAs end in a highly conserved stem-loop structure followed by ACCCA. Previous studies have suggested that the stem-loop binding protein (SLBP) is the only protein binding this region. Using RNA affinity purification, we identified a second protein, designated 3'hExo, that contains a SAP and a 3' exonuclease domain and binds the same sequence. Strikingly, 3'hExo can bind the stem-loop region both separately and simultaneously with SLBP. Binding of 3'hExo requires the terminal ACCCA, whereas binding of SLBP requires the 5' side of the stem-loop region. Recombinant 3'hExo degrades RNA substrates in a 3'-5' direction and has the highest activity toward the wild-type histone mRNA. Binding of SLBP to the stem-loop at the 3' end of RNA prevents its degradation by 3'hExo. These features make 3'hExo a primary candidate for the exonuclease that initiates rapid decay of histone mRNA upon completion and/or inhibition of DNA replication. PMID- 14536071 TI - Separate insertion and deletion subcomplexes of the Trypanosoma brucei RNA editing complex. AB - The Trypanosoma brucei editosome catalyzes the maturation of mitochondrial mRNAs through the insertion and deletion of uridylates and contains at least 16 stably associated proteins. We examined physical and functional associations among these proteins using three different approaches: purification of complexes via tagged editing ligases TbREL1 and TbREL2, comprehensive yeast two-hybrid analysis, and coimmunoprecipitation of recombinant proteins. A purified TbREL1 subcomplex catalyzed precleaved deletion editing in vitro, while a purified TbREL2 subcomplex catalyzed precleaved insertion editing in vitro. The TbREL1 subcomplex contained three to four proteins, including a putative exonuclease, and appeared to be coordinated by the zinc finger protein TbMP63. The TbREL2 subcomplex had a different composition, contained the TbMP57 terminal uridylyl transferase, and appeared to be coordinated by the TbMP81 zinc finger protein. This study provides insight into the molecular architecture of the editosome and supports the existence of separate subcomplexes for deletion and insertion editing. PMID- 14536072 TI - Ribosomal proteins S12 and S13 function as control elements for translocation of the mRNA:tRNA complex. AB - Translocation of the mRNA:tRNA complex through the ribosome is promoted by elongation factor G (EF-G) during the translation cycle. Previous studies established that modification of ribosomal proteins with thiol-specific reagents promotes this event in the absence of EF-G. Here we identify two small subunit interface proteins S12 and S13 that are essential for maintenance of a pretranslocation state. Omission of these proteins using in vitro reconstitution procedures yields ribosomal particles that translate in the absence of enzymatic factors. Conversely, replacement of cysteine residues in these two proteins yields ribosomal particles that are refractive to stimulation with thiol modifying reagents. These data support a model where S12 and S13 function as control elements for the more ancient rRNA- and tRNA-driven movements of translocation. PMID- 14536073 TI - Cotranslational protein integration into the ER membrane is mediated by the binding of nascent chains to translocon proteins. AB - During cotranslational protein integration into the ER membrane, each transmembrane (TM) segment moves laterally through the translocon to reach the lipid bilayer. Photocrosslinking studies reveal that a particular surface of each nascent chain TM alpha helix and signal-anchor sequence always faces Sec61alpha in the translocon. This nonrandom and TM sequence-dependent positioning reveals that each TM segment makes specific contacts with Sec61alpha and is retained at a fixed location within the translocon, observations that are best explained by the binding of each TM sequence to a translocon protein(s). Since TM sequence hydrophobicity does not correlate with its rate of release from the translocon, nascent chain movement through the translocon appears to be mediated primarily by protein-protein interactions rather than hydrophobic nascent chain-phospholipid interactions. PMID- 14536074 TI - A despecialization step underlying evolution of a family of serine proteases. AB - In the trypsin superfamily of serine proteases, non-trypsin-like primary specificities have arisen in only two monophyletic descendent subbranches. We have recreated an ancestor to one of these subbranches (granzyme) using phylogenetic inference, gene synthesis, and protein expression. This ancestor has two unusual properties. First, it has broad primary specificity encompassing the entire repertoire of novel primary specificities found in its descendents. Second, unlike extant members that have narrow primary specificities, the ancestor exhibits tolerance to mutational changes in primary specificity conferring residues-that is, structural plasticity. Molecular modeling and mutagenesis studies indicate that these unusual properties are due to a particularly wide substrate binding pocket. These two crucial properties of the ancestor not only distinguish it from its extant descendents but also from the trypsin-like proteases that preceded it. This indicates that a despecialization step, characterized by broad specificity and structural plasticity, underlies evolution of new primary specificities in this protease superfamily. PMID- 14536075 TI - Flexible linkers leash the substrate binding domain of SspB to a peptide module that stabilizes delivery complexes with the AAA+ ClpXP protease. AB - SspB dimers bind proteins bearing the ssrA-degradation tag and stimulate their degradation by the ClpXP protease. Here, E. coli SspB is shown to contain a dimeric substrate binding domain of 110-120 N-terminal residues, which binds ssrA tagged substrates but does not stimulate their degradation. The C-terminal 40-50 residues of SspB are unstructured but are required for SspB to form substrate delivery complexes with ClpXP. A synthetic peptide containing the 10 C-terminal residues of SspB binds ClpX, stimulates its ATPase activity, and prevents SspB mediated delivery of GFP-ssrA for ClpXP degradation. This tripartite structure- an ssrA-tag binding and dimerization domain, a flexible linker, and a short peptide module that docks with ClpX--allows SspB to deliver tagged substrates to ClpXP without interfering with their denaturation or degradation. PMID- 14536076 TI - Structure of a delivery protein for an AAA+ protease in complex with a peptide degradation tag. AB - Substrate selection by AAA+ ATPases that function to unfold proteins or alter protein conformation is often regulated by delivery or adaptor proteins. SspB is a protein dimer that binds to the ssrA degradation tag and delivers proteins bearing this tag to ClpXP, an AAA+ protease, for degradation. Here, we describe the structure of the peptide binding domain of H. influenzae SspB in complex with an ssrA peptide at 1.6 A resolution. The ssrA peptides are bound in well-defined clefts located at the extreme ends of the SspB homodimer. SspB contacts residues within the N-terminal and central regions of the 11 residue ssrA tag but leaves the C-terminal residues exposed and positioned to dock with ClpX. This structure, taken together with biochemical analysis of SspB, suggests mechanisms by which proteins like SspB escort substrates to AAA+ ATPases and enhance the specificity and affinity of target recognition. PMID- 14536077 TI - Targeted delivery of an ssrA-tagged substrate by the adaptor protein SspB to its cognate AAA+ protein ClpX. AB - In the bacterial cytosol, degradation of ssrA-tagged proteins is primarily carried out by the proteolytic machine ClpXP in a process which is stimulated by a ClpX-specific adaptor protein, SspB. Here we elucidate the steps required for binding and transfer of ssrA-tagged substrates from SspB to ClpX. The N-terminal region of SspB is essential for its interaction with ssrA-tagged substrates, while a short conserved region at the C terminus of SspB interacts specifically with the N domain of ClpX. A single point mutation within the conserved C terminal region of SspB is sufficient to abolish the SspB-mediated degradation of ssrA-tagged proteins by ClpXP. We propose that this region represents a common motif for the recognition of ClpX as the C-terminal region of SspB shares considerable homology with the other ClpX-specific adaptor protein, RssB. Through docking of SspB to the N-terminal domain of ClpX, the substrate is delivered to the substrate binding site in ClpX. PMID- 14536078 TI - Multisite phosphorylation by Cdk2 and GSK3 controls cyclin E degradation. AB - Autophosphorylation-triggered ubiquitination has been proposed to be the major pathway regulating cyclin E protein abundance: phosphorylation of cyclin E on T380 by its associated CDK allows binding to the receptor subunit, Fbw7, of the SCFFbw7 ubiquitin ligase. We have tested this model in vivo and found it to be an inadequate representation of the pathways that regulate cyclin E degradation. We show that assembly of cyclin E into cyclin E-Cdk2 complexes is required in vivo for turnover by the Fbw7 pathway; that Cdk2 activity is required for cyclin E turnover in vivo because it phosphorylates S384; that phosphorylation of T380 in vivo does not require Cdk2 and is mediated primarily by GSK3; and that two additional phosphorylation sites, T62 and S372, are also required for turnover. Thus, cyclin E turnover is controlled by multiple biological inputs and cannot be understood in terms of autophosphorylation alone. PMID- 14536079 TI - Bmi-1 regulation of INK4A-ARF is a downstream requirement for transformation of hematopoietic progenitors by E2a-Pbx1. AB - Loss-of-function alterations of INK4A are commonly observed in lymphoid malignancies, but are consistently absent in pre-B cell leukemias induced by the chimeric oncoprotein E2a-Pbx1 created by t(1;19) chromosomal translocations. We report here that experimental induction of E2a-Pbx1 enhances expression of BMI-1, a lymphoid oncogene whose product functions as a transcriptional repressor of the INK4A-ARF tumor suppressor locus. Bmi-1-deficient hematopoietic progenitors are resistant to transformation by E2a-Pbx1; however, the requirement for Bmi-1 is alleviated in cells deficient for both Bmi-1 and INK4A-ARF. Furthermore, the adverse effects of E2a-Pbx1 on pre-B cell survival and differentiation are partially bypassed by forced expression of p16(Ink4a). These results link E2a Pbx1 with Bmi-1 on an oncogenic pathway that is likely to play a role in the pathogenesis of human lymphoid leukemias through downregulation of the INK4A-ARF gene. PMID- 14536080 TI - Retrograde signaling is regulated by the dynamic interaction between Rtg2p and Mks1p. AB - Activation of retrograde signaling (RS) by mitochondrial dysfunction or by inhibition of TOR kinases in yeast results in nuclear accumulation of the transcription factors, Rtg1p and Rtg3p. This process requires Rtg2p, a novel cytoplasmic protein with an N-terminal ATP binding domain. We show that Rtg2p controls RS by reversibly binding a negative regulator, Mks1p. The inhibitory form of Mks1p is phosphorylated and complexed with the 14-3-3 proteins, Bmh1p and Bmh2p, which are also negative regulators of RS. A hypophosphorylated form of Mks1p bound to Rtg2p is inactive. Point mutations in the Rtg2p ATP binding domain simultaneously block RS and Mks1p-Rtg2p interaction. We propose that activation of RS via mitochondrial dysfunction and TOR inhibition intersect at the Rtg2p Mks1p switch. PMID- 14536081 TI - TORCs: transducers of regulated CREB activity. AB - The cAMP responsive factor CREB stimulates gene expression, following its phosphorylation at Ser133, via recruitment of the coactivator CBP. In certain cell types, CREB also functions as a constitutive activator, although the underlying mechanisms are not understood. Here, we characterize a conserved family of coactivators, designated TORCs, for Transducers of Regulated CREB activity, that enhances CRE-dependent transcription via a phosphorylation independent interaction with the bZIP DNA binding/dimerization domain of CREB. TORC recruitment does not appear to modulate CREB DNA binding activity, but rather enhances the interaction of CREB with the TAF(II)130 component of TFIID following its recruitment to the promoter. Remarkably, in certain mucoepidermoid carcinomas, a chromosomal translocation fuses the CREB binding domain of TORC1 to the Notch coactivator Mastermind (MAML2). As expression of the TORC1-MAML2 chimera strongly induced target gene expression via CREB, our results reveal a mechanism by which CREB stimulates transcription in normal and transformed cells. PMID- 14536082 TI - A mutant form of MeCP2 protein associated with human Rett syndrome cannot be displaced from methylated DNA by notch in Xenopus embryos. AB - MeCP2 is a DNA binding protein that represses transcription of methylated genes in vitro, but the endogenous function of MeCP2 in vivo is unclear. Here, we demonstrate that in Xenopus laevis embryos MeCP2 is a partner of the SMRT corepressor complex that regulates the expression of a neuronal repressor xHairy2a in differentiating neuroectoderm. The MeCP2/SMRT complex is bound to the promoter of the silenced xHairy2a gene and is displaced upon activation by the Notch intracellular domain (NICD). A truncated form of MeCP2 (R168X) found in patients with Rett syndrome cannot interact with the SMRT complex or fully activate xHairy2a during primary neurogenesis. This disruption of MeCP2 activity results in abnormal patterning of primary neurons during neuronal differentiation. Our results support a model whereby the dynamic association of MeCP2 with methylated DNA and the SMRT complex regulates a gene involved in cell fate decisions during primary neurogenesis in Xenopus. PMID- 14536083 TI - Binding of TFIIB to RNA polymerase II: Mapping the binding site for the TFIIB zinc ribbon domain within the preinitiation complex. AB - RNA polymerase II (Pol II) is recruited to promoters by interaction with general transcription factors. The zinc ribbon domain of the general factor TFIIB is essential for Pol II recruitment. Site-specific photocrosslinking and directed hydroxyl radical probing were used to map the location of the TFIIB zinc ribbon domain on Pol II within the transcription preinitiation complex (PIC). These results, along with mutational analysis, suggest that in the PIC, the TFIIB ribbon domain interacts with a surface of the Pol II Dock domain where it overlaps the RNA exit point. This surface is best conserved in polymerases that require a TFIIB-like factor. Our results suggest a general mechanism for interaction of TFIIB-like factors and RNA polymerases and a mechanism for the function of the ribbon domain. PMID- 14536084 TI - Crystal structure and functional analysis of a nucleosome recognition module of the remodeling factor ISWI. AB - Energy-dependent nucleosome remodeling emerges as a key process endowing chromatin with dynamic properties. However, the principles by which remodeling ATPases interact with their nucleosome substrate to alter histone-DNA interactions are only poorly understood. We have identified a substrate recognition domain in the C-terminal half of the remodeling ATPase ISWI and determined its structure by X-ray crystallography. The structure comprises three domains, a four-helix domain with a novel fold and two alpha-helical domains related to the modules of c-Myb, SANT and SLIDE, which are linked by a long helix. An integrated structural and functional analysis of these domains provides insight into how ISWI interacts with the nucleosomal substrate. PMID- 14536085 TI - Structural basis for histone and phosphohistone binding by the GCN5 histone acetyltransferase. AB - Distinct posttranslational modifications on histones occur in specific patterns to mediate certain chromosomal events. For example, on histone H3, phosphorylation at Ser10 can enhance GCN5-mediated Lys14 acetylation to promote transcription. To gain insight into the mechanism underlying this synergism, we determined the structure of Tetrahymena GCN5 (tGCN5) and coenzyme A (CoA) bound to unmodified and Ser10-phosphorylated 19 residue histone H3 peptides (H3p19 and H3p19Pi, respectively). The tGCN5/CoA/H3p19 structure reveals that a 12 amino acid core sequence mediates extensive contacts with the protein, providing the structural basis for substrate specificity by the GCN5/PCAF family of histone acetyltransferases. Comparison with the tGCN5/CoA/H3p19Pi structure reveals that phospho-Ser10 and Thr11 mediate significant histone-protein interactions, and nucleate additional interactions distal to the phosphorylation site. Functional studies show that histone H3 Thr11 is necessary for optimal transcription at yGcn5-dependent promoters requiring Ser10 phosphorylation. Together, these studies reveal how one histone modification can modulate another to affect distinct transcriptional signals. PMID- 14536086 TI - mAM facilitates conversion by ESET of dimethyl to trimethyl lysine 9 of histone H3 to cause transcriptional repression. AB - Methylation of histone tails plays an important role in chromatin structure and function. Previously, we reported that ESET/SETDB1 is a histone methyltransferase (HMTase). Here, we show that SETDB1 tightly associates with the human homolog of mAM, a murine ATFa-associated factor. Although recombinant ESET can methylate lysine 9 of histone H3 (H3-K9), its activity is severely compromised when compared to that of the ESET/mAM complex. mAM stimulates ESET enzymatic activity by increasing the Vmax and decreasing the Km. Importantly, mAM facilitates the ESET-dependent conversion of dimethyl H3-K9 to the trimethyl state both in vitro and in vivo. Chromatin-based transcription and ChIP analyses demonstrate that mAM enhances ESET-mediated transcriptional repression in a SAM-dependent manner, and this repression correlates with H3-K9 trimethylation at the promoter. Thus, our studies establish that promoter H3-K9 trimethylation is the cause of transcriptional repression and that mAM/hAM facilitates conversion of H3-K9 dimethyl to trimethyl by ESET/SETDB1. PMID- 14536087 TI - V(D)J recombination and RAG-mediated transposition in yeast. AB - Antigen receptor genes are assembled during lymphoid development by a specialized recombination reaction normally observed only in cells of the vertebrate immune system. Here, we show that expression in Saccharomyces cerevisiae of murine RAG1 and RAG2, the lymphoid-specific components of the V(D)J recombinase, is sufficient to induce V(D)J cleavage and rejoining in this lower eukaryote. The RAG proteins cleave recombination substrates introduced into yeast cells, generating signal ends that can be joined to form signal joints. These signal joints are precise, as in mammalian cells, and their formation is dependent on a yeast nonhomologous end-joining protein, the XRCC4 homolog LIF1. Moreover, joining of SmaI-generated blunt ends is generally imprecise in the yeast strain used here, suggesting that the RAG proteins influence signal-end joining. Cleaved signal ends are also transposed into new sites in DNA, allowing RAG-induced transposition to be studied in vivo. PMID- 14536088 TI - C-terminal deletion of AID uncouples class switch recombination from somatic hypermutation and gene conversion. AB - Class-switch recombination (CSR), somatic hypermutation (SHM), and antibody gene conversion are distinct DNA modification reactions, but all are initiated by activation-induced cytidine deaminase (AID), an enzyme that deaminates cytidine residues in single-stranded DNA. Here we describe a mutant form of AID that catalyzes SHM and gene conversion but not CSR. When expressed in E. coli, AID(delta189-198) is more active in catalyzing cytidine deamination than wild type AID. AID(delta189-198) also promotes high levels of gene conversion and SHM when expressed in eukaryotic cells, but fails to induce CSR. These results underscore an essential role for the C-terminal domain of AID in CSR that is independent of its cytidine deaminase activity and that is not required for either gene conversion or SHM. PMID- 14536089 TI - Transcription within a functional human centromere. AB - Recent data in yeast and Drosophila suggest a domain-like centromere structure with a modified chromatin core and flanking regions of heterochromatin. We have analyzed a functional human centromere and defined a region of increased chromosome scaffold/matrix attachment that overlaps three other distinct and nonoverlapping domains for constitutive centromere proteins CENP-A and CENP-H, and heterochromatin protein HP1. Transcriptional competency is intact throughout the S/MAR-enriched region and within the CENP-A- and CENP-H-associated chromatin. These results provide insights into the relationship between centromeric chromatin and transcriptional competency in vivo, highlighting the permissibility of transcription within the constitutively modified, nonheterochromatic chromatin of a functional eukaryotic centromere. PMID- 14536090 TI - The yeast G protein alpha subunit Gpa1 transmits a signal through an RNA binding effector protein Scp160. AB - In yeast Saccharomyces cerevisiae the G protein betagamma subunits (Ste4/Ste18) have long been known to transmit the signal required for mating. Here we demonstrate that GTPase-deficient mutants of Galpha (Gpa1) directly activate the mating response pathway. We also show that signaling by activated Gpa1 requires direct coupling to an RNA binding protein Scp160. These findings suggest an additional role for Gpa1 and reveal Scp160 as a component of the mating response pathway in yeast. PMID- 14536091 TI - A slow RNA polymerase II affects alternative splicing in vivo. AB - Changes in promoter structure and occupation have been shown to modify the splicing pattern of several genes, evidencing a coupling between transcription and alternative splicing. It has been proposed that the promoter effect involves modulation of RNA pol II elongation rates. The C4 point mutation of the Drosophila pol II largest subunit confers on the enzyme a lower elongation rate. Here we show that expression of a human equivalent to Drosophila's C4 pol II in human cultured cells affects alternative splicing of the fibronectin EDI exon and adenovirus E1a pre-mRNA. Most importantly, resplicing of the Hox gene Ultrabithorax is stimulated in Drosophila embryos mutant for C4, which demonstrates the transcriptional control of alternative splicing on an endogenous gene. These results provide a direct proof for the elongation control of alternative splicing in vivo. PMID- 14536092 TI - Patenting human gene therapy. PMID- 14536093 TI - Antifreeze proteins and their potential use in frozen foods. AB - Antifreeze proteins (AFPs) are proteins that have the ability to modify the growth of ice, resulting in the stabilization of ice crystals over a defined temperature range and in the inhibition of the recrystallization of ice. AFPs are found in a wide range of organisms, including bacteria, fungi, plants, invertebrates and fish. Moreover, multiple forms of AFPs are synthesized within each organism. As a result, it should be possible to select an AFP with appropriate characteristics and a suitable level of activity for a particular food product. Antifreeze proteins may improve the quality of foods that are eaten while frozen by inhibiting recrystallization and maintaining a smooth texture. In foods that are frozen only for preservation, AFPs may inhibit recrystallization during freezing, storage, transport and thawing, thus preserving food texture by reducing cellular damage and also minimizing the loss of nutrients by reducing drip. Antifreeze proteins are naturally present in many foods consumed as part of the human diet. However, AFPs may be introduced into other food products either by physical processes, such as mixing and soaking, or by gene transfer. PMID- 14536094 TI - The impact of Arabidopsis research on plant biotechnology. AB - Arabidopsis thaliana, a small annual weed belonging to the mustard family, has become a widely used model in plant genetic research. It has a small genome, short life cycle, and is easy to mutagenize. Identification of genes based on phenotype alone, often a rather difficult part of molecular genetic research, is easiest in this plant. Laboratories working on the "model" plant Arabidopsis thaliana have created a network for sharing resources and ideas, so progress has been rapid. The importance of this plant to biotechnology is that genes isolated from Arabidopsis can be used to find their homologs in crop plants. Likewise, fundamental mechanisms can be understood in a model plant, and applied in crop plants. PMID- 14536095 TI - The development of Azospirillum as a commercial inoculant for improving crop yields. AB - Bacteria of the genus Azospirillum are nitrogen-fixing organisms that live in close association with plants in the rhizosphere. The Azospirillum-plant association leads to the enhanced development and yield of different host plants under appropriate growth conditions. This increase in yield is attributed mainly to an improvement in root development, an increase in the rate of water and mineral uptake by roots, and to a lesser extent, biological N(2) fixation. Worldwide data accumulated in the field over the past 20 years indicates that Azospirillum is capable of promoting the yield of agriculturally important crops in different soils and climatic regions. A.brasilense shows both chemotaxis and chemokinesis in response to temporal gradients of different chemoeffectors, thereby increasing the chance of root-bacterial interactions. Phytohormones synthesized by Azospirillum influence the host root respiration rate, metabolism and root proliferation and hence better the mineral and water uptake in inoculated plants. Positive effects of combined inoculation with Rhizobium have been reported for different legumes and were related to the favorable influence of Azospirillum on the nodule number, plant development, dry weight, and N(2) fixation. Additionally, A. brasilense produces the reserve material polyhydroxybutyrate comprising up to 70% of the cell dry weight This substance has received much attention recently as it can be extracted and formed into a biodegradable thermoplastic. PMID- 14536096 TI - Plant cell and tissue culture: alternatives for metabolite production. AB - Plant cell culture systems represent a potential renewable source of valuable medicinals, flavours, essences and colourants that cannot be produced by microbial cells or chemical syntheses. However, only a few cultures produce these compounds in commercially useful amounts. The low productivities are associated with our poor understanding of the biochemistry of these systems. Recent advances in molecular biology, enzymology, physiology and fermentation technology of plant cell cultures suggest that these systems will become a viable source of important natural products. This review examines the sate of the art of production of medicinal plant secondary metabolites by plant cell cultures. PMID- 14536097 TI - The impact of biotechnology on hyphomycetous fungal insect biocontrol agents. AB - The potential for the control of insect pests by entomopathogenic fungi has been touted for decades, if not centuries. Only recently have advances in biotechnology provided the tools for indepth analysis of the mechanisms involved in pathogenesis and host death at the molecular level. This review outlines the current state of knowledge regarding the mode of infection and targets several key components that are amenable to improvement via biotechnology. Realization of the considerable economic potential of fungal bioinsecticides can occur only through a combined and coordinated effort involving fundamental science, formulation technology and field applications. PMID- 14536098 TI - Process-scale disruption of microorganisms. AB - Common hosts for the large-scale manufacture of biological products, such as Escherichia coli and Saccharomyces cerevisiae, do not excrete products to the medium. Effective techniques for cell disruption are therefore required. These include physical, chemical, enzymatic and mechanical methods. Mechanical methods such as bead milling, high-pressure homogenization, and microfluidization are preferred. However, gentler, specific methods are receiving increasing attention particularly when used in combination to synergistically exploit their different specificities. Benefits can also be derived by integrating product release and recovery. In all cases it is essential to consider the interaction of the disruption operation with downstream units and to clearly demonstrate the cost benefits of alternative strategies. PMID- 14536367 TI - Factors affecting the genetic engineering of plants by microprojectile bombardment. AB - Since its development in the mid-1980s, microprojectile bombardment has been widely employed as a method for direct gene transfer into a wide range of plants, including the previously difficult-to-transform monocotyledonous species. Although the numerous instruments available for microprojectile-mediated gene delivery and their applications have been widely discussed, less attention has been paid to the critical factors which affect the efficiency of this method of gene delivery. In this review we do not wish to describe the array of devices used for microprojectile delivery or their uses which have already been definitively described, but instead wish to report on research developments investigating the factors which affect microprojectile-mediated transformation of plants. PMID- 14536368 TI - Foreign gene delivery into monocotyledonous species. AB - Monocotyledonous plants are generally more recalcitrant to genetic transformation than dicotyledonous species. The absence of reliable Agrobacterium-mediated transformation methods and the difficulties associated with the culture of monocotyledonous tissues in vitro are mainly responsible for this situation. Until recently, the genetic transformation of monocotyledons was essentially performed by direct transfer of DNA into regenerable protoplasts or intact cells cultured in vitro, via polyethylene glycol treatment, electroporation or particle bombardment. Since 1990, the use of particle gun technology has revolutionized the genetic engineering of monocotyledonous species, allowing transformation to be more independent of the in vitro culture requirements. Today, at least one genotype of each major monocotyledonous crop species, including cereals, can be genetically transformed. PMID- 14536369 TI - Plant biotechnology for crop improvement. AB - The typical crop improvement cycle takes 10-15 years to complete and includes germplasm manipulations, genotype selection and stabilization, variety testing, variety increase, proprietary protection and crop production stages. Plant tissue culture and genetic engineering procedures that form the basis of plant biotechnology can contribute to most of these crop improvement stages. This review provides an overview of the opportunities presented by the integration of plant biotechnology into plant improvement efforts and raises some of the societal issues that need to be considered in their application. PMID- 14536921 TI - Multifactional glucanases. AB - Diverse functional roles of multifunctional proteins arise from either their independent functional domains or dual activities mediated through a single active site. Presence of multifunctional proteins significantly enhances the metabolic efficiency of a cell. Microorganisms utilising complex substrates with extensive microheterogeneities, such as carbohydrates evolved batteries of multifunctional glucanases, facilitating parsimonious utilisation of these substrates. Various attempts have since been made to artificially construct these glucanases. Analysis of information on various glucanases would be helpful in understanding the evolutionary interrelationship between this class of enzymes and will give an insight into the structural features controlling different unrelated activities. This review examines the genesis, evolution and structural features of multifunctional glucanases. PMID- 14536922 TI - Patentable invention in biotechnology. AB - Biotechnology is a rapidly advancing field of technology which offers many benefits to society. It is perceived that an important part of maintaining the momentum, and stimulating further advances, is to protect inventions in biotechnology, where appropriate, with a patent. Here, from the perspective of invention in biotechnology, we examine in brief, the critical elements of proper subject-matter, novelty, utility, non-obviousness and sufficiency of disclosure, as requirements of patentable invention for both the United States and Canada. Topical issues which have arisen in respect of these elements are also canvassed briefly. As will be seen, Canadian Patent Law, while still unresolved with respect to the patenting of higher life forms, is in other respects in step with the pro-patent model of the United States. PMID- 14536923 TI - Biodegradation of chlorinated phenolic compounds. AB - Chlorophenolic compounds are generated from a number of industrial manufacturing processes including pulp and paper manufacture. These compounds are found to be toxic and recalcitrant and hence their discharge into the environment must be regulated. Slow and partial degradation of chlorophenols under aerobic and anaerobic natural environment has been observed. Aerobic biodegradation of chlorophenols proceeds through the formation of catechols while under anaerobic conditions, reductive dehalogenation is the preferred metabolic pathway. Number and position of chlorine substituents on the phenolic ring has influence on the rate and extent of biodegradation of chlorophenols. In engineered systems, acclimatization of biomass to chlorophenols markedly enhances the biodegradation ability by reducing the initial lag phase and by countering inhibition. Partial removal of chlorophenols between 40-60% is usually observed in aerobic and anaerobic processes. Removal can be enhanced by a combination of aerobic and anaerobic operations. PMID- 14536924 TI - The zebrafish as a model system in developmental, toxicological and transgenic research. AB - The zebrafish has long been used as a model system in fisheries biology and toxicology. More recently, it has also become the focus of a major research effort into understanding the molecular and cellular events which dictate the development of vertebrate embryos. As well, the zebrafish has proven attractive in studies examining the factors which affect the creation of transgenic fish and the expression of transgenes. The advances which have been made in these areas have firmly established this small aquarium fish as a major model system in biological and biotechnological research. PMID- 14537102 TI - 2-Deoxy-2-[18F]fluoro-D-glucose positron emission tomography in Alzheimer's diagnosis: time for technology transfer. PMID- 14537103 TI - From thallium scan to molecular imaging. AB - The diagnostic and prognostic evaluation of cardiovascular diseases has been improved considerably by the application of imaging procedures. Among many, scintigraphic procedures have emerged as important diagnostic tools to assess extent, severity, and prognosis in patients with coronary artery disease (CAD). For more than the last 30 years, however, the application of perfusion imaging has been extended to allow the combined evaluation of perfusion, perfusion reserve, and ventricular function. With positron emission tomography (PET), quantitative assessment of perfusion has become possible. In combination with pharmacological stress agents, the coronary flow reserve (CFR) can be quantitatively assessed as an early marker of endothelial dysfunction. PET in combination with metabolic tracers has added the evaluation of cardiac substrate metabolism, which has become an important clinical marker for ischemically jeopardized myocardium. The PET information is widely considered as the gold standard for tissue viability in the management of patients with advanced CAD and impaired left ventricular function (LVF). New tracer approaches include the assessment of cardiac innervation, which plays an increasingly recognized role in the pathophysiology of cardiac diseases. Radiolabeled catecholamine analogues provide visualization of sympathetic nerve terminals that are functionally altered in patients with diabetes mellitus and cardiomyopathy. In addition, this scintigraphic information allows the monitoring of physiological processes such as reinnervation of the transplanted heart. New methods, such as imaging of apoptosis and gene expression, are of interest in cardiology. Combining the therapeutic gene with a reporter gene, the transfection of cardiac tissue can be monitored noninvasively. First results employing the herpes simplex virus thymidine kinase reporter gene (HSV1-tk) are encouraging and represent an attractive approach for the use of PET imaging in the control of cardiac gene therapy. PMID- 14537104 TI - Quantitative evaluation of 2-deoxy-2-[18F] fluoro-D-glucose uptake in hepatic metastases with combined PET-CT: iterative reconstruction with CT attenuation correction versus filtered back projection with 68Germanium attenuation correction. AB - PURPOSE: Iterative reconstruction (IR) is a statistical reconstruction method that may be influenced by high background activity such as in the liver. Recently developed combined positron emission tomography (PET) and computerized tomography (CT) instrumentation utilizes CT attenuation correction that may also differ compared to 68Germanium (68Ge) segmented attenuation correction (IR SAC). Quantitative differences could affect the clinical interpretation of metastatic foci and subsequent response to therapy. The purpose of this study is to characterize potential quantitative differences specifically in hepatic metastases. PROCEDURES: Thirty-three metastatic liver lesions in 23 patients were evaluated. Whole-body 2-deoxy-2[18F]fluoro-D-glucose(FDG) PET images were obtained on a dedicated PET-CT device and reconstructed with IR and CT measured attenuation correction (IR CT MAC), IR and 68Ge segmented attenuation correction, and also with filtered back-projection and segmented attenuation correction (FBP SAC). Regions of interest over the liver lesions (L) and liver background (B) were drawn on FBP SAC images and superimposed on the co-registered IR CT MAC and IR SAC images for comparison. To identify the individual effects of IR and CT attenuation correction, IR SAC was first compared to FBP SAC, followed by a comparison of IR CT MAC and IR SAC. Differences were expressed as bias (%) and compared to FBP SAC. RESULTS: Compared to FBP SAC, IR SAC showed significantly lower liver lesion (mean bias -7.1%; P < 0.00001), significantly higher liver background (mean bias -3.5%; P = 0.005), and significantly lower L/B ratio (mean bias -10.1%; P < 0.00001). Compared to IR SAC, the IR CT MAC showed small but not statistically significant increases in liver lesion (mean bias 3.0%; P = 0.1) and liver background (mean bias 4.3%; P = 0.09), and no significant difference in L/B ratio (mean bias -1.3%; P = 0.5). The overall effect of IR CT MAC compared to FBP SAC was a significant decrease in liver lesion (mean bias -4.2%; P = 0.002), significant increase in liver background (mean bias 8.7%; P = 0.002), and significantly lower L/B (mean bias -11.1%; P < 0.00001) compared to FBP SAC. CONCLUSION: Compared to FBP, IR resulted in significantly lower mean hepatic tumor activity, higher mean liver background activity, and lower L/B activity, but these differences were modest. These differences were similarly seen in the CT corrected IR CT MAC when compared to FBP SAC. CT attenuation correction did not result in significant differences when compared to 68Ge correction. Although the differences in IR CT MAC were modest, consistent reconstruction methods are important to decrease the measurement variability and improve reproducibility. PMID- 14537106 TI - A fully automated one pot synthesis of 9-(4-[18F]fluoro-3-hydroxymethylbutyl) guanine for gene therapy studies. AB - PURPOSE: To develop a new fully-automated method for the synthesis of 9-(4 [18F]fluoro-3-hydroxymethylbutyl)guanine ([18F]FHBG) amenable for its routine use in gene therapy monitoring studies. PROCEDURES: A nuclear interface commercial synthesizer was substantially modified and adapted to the synthesis of the referred compound. After the fluorination reaction of the tosylate precursor, the intermediate product was purified by Solid Phase Extraction (SPE) before the hydrolysis. The final product was purified by semi-preparative high performance liquid chromatography (HPLC). RESULTS: [18F]FHBG was obtained in 10-15% yield in 65 minutes including HPLC purification. The radiotracer was > 99% chemically and radiochemically pure, sterile and free from pyrogens. The synthesized compound was shown to accumulate in thymidine kinase (tk) expressing cells both in cell culture, and in laboratory animals infected with an adenoviral vector containing the herpes simplex virus type 1 thymidine kinase (HSV1-tk) gene. CONCLUSIONS: This new procedure facilitates the compliance with the applicable regulatory guidelines for positron emission tomography (PET) radiopharmaceuticals and will assist the clinical application of [18F]FHBG-PET as a noninvasive way to monitor gene therapy in humans. PMID- 14537105 TI - Contamination levels in blood samples drawn from the injection intravenous line. AB - PURPOSE: Securing two intravenous lines, one for injection and one for blood sampling, can be nearly impossible in compromised patients, therefore, a need exists to quantify the potential error when simplified techniques are employed. METHOD: Two venous catheters were placed. 2-deoxy-2-[18F]fluoro-glucose (FDG) was infused through one of the catheters. Venous blood samples were drawn from each line. Triplicate aliquots of plasma were analyzed in duplicate. RESULTS: Concentrations from the infusion line were 2.0% higher than the concentrations from the noninfusion line. The average error was 3.3%, 2.0%, and 0.7% higher for the first, second, and third samples, respectively. CONCLUSIONS: Blood sampling through the infusion catheter is a viable alternative to the placement of separate venous catheters. Sampling from the injection catheter, even with tubing flush and replacement, will potentially incur small (generally < 10%) over estimations in concentration in initial samples. Subsequent sampling reduces the error to essentially zero by the third sample. PMID- 14537107 TI - Positron emission tomography and drug discovery: contributions to the understanding of pharmacokinetics, mechanism of action and disease state characterization. AB - As an imaging modality, positron emission tomography (PET) provides unique quantitative in vivo information of value to drug discovery studies. These non invasive studies span the pharmacokinetic/pharmacodynamic evaluation of potential drug candidates, receptor occupancy as an important determinant of efficacy, the pharmacological characterization of potential mechanisms of action, and the biological characterization of disease with well-characterized PET ligands. PET techniques are also being applied to the assessment of gene-level activities and the longitudinal evaluation of disease progression and therapeutic intervention. As the availability of PET scanners, cyclotrons, and specific PET ligands grows, the techniques highlighted in this review will become central to target validation, drug candidate selection, pharmacokinetic characterization, and clinical evaluation. PMID- 14537108 TI - Positron emission tomography and epilepsy. AB - PURPOSE: This review examines the current role of positron emission tomography (PET) in the investigation and management of patients with epilepsy. PROCEDURES: A literature review utilizing MEDLINE(R) and other sources was undertaken. For the comparison of the accuracy of PET with magnetic resonance imaging (MRI) for seizure focus localization, only publications since 1994 were examined. Individual patient data was tabulated to provide figures for seizure focus localization rates for different types of focal epilepsy and the prognostic value of PET findings for epilepsy surgery outcome. RESULTS: The majority of PET studies used 2-deoxy-2-[18F]fluoro-D-glucose (FDG). The epileptogenic sites typically show reduced FDG uptake (hypometabolism). In patients with intractable temporal lobe epilepsy (TLE), unilateral temporal lobe hypometabolism (UTH) corresponding to the seizure focus was seen in 86% of patients. In the same population, MRI demonstrated relevant abnormalities in 76%. UTH contralateral to the seizure focus was rarely seen (3%). Following temporal lobectomy, 86% of patients with ipsilateral UTH had a good outcome. When MRI was normal, UTH predicted a good outcome in 82%. Fifty percent with bitemporal hypometabolism had independent bilateral foci, and in those who proceeded to surgery only 50% had a good result. In extratemporal epilepsy, hypometabolism relevant to the focus was seen in 67% but, as in TLE, it was often more extensive than pathological abnormality. Recently evidence of a role for 11C-Flumazenil has emerged with reduced binding in the primary epileptogenic site. 11C-Flumazenil abnormalities appear more restricted to abnormal cortex and may be a better guide to the extent of resection required for surgical success. CONCLUSIONS: FDG-PET has a key role in the evaluation of patients with intractable partial epilepsy, particularly when surgery is a treatment option. Development and application of more specific biochemical probes may further improve the clinical value of PET for the understanding and treatment of epilepsy. PMID- 14537109 TI - History of the first synthesis of 2-deoxy-2-fluoro-D-glucose the unlabeled forerunner of 2-deoxy-2-[18F]fluoro-D-glucose. AB - The history of the first successful synthesis of 2-deoxy-2-fluoro-D-glucose (19FDG) is described. In many aspects, this substance imitates the behavior of naturally occurring glucose. For example, it is transported into the cells and is converted to the corresponding 6-phosphate by the enzyme hexokinase in a manner similar to glucose. Due to the presence of the fluorine atom at C-2, however, this phosphate derivative does not undergo further glycolysis but is metabolically trapped in the cell. Thanks to these properties, eight years after the synthesis of 19FDG, its 18F-labeled derivative was successfully used with positron emission tomography (PET). PMID- 14537110 TI - 2-Deoxy-2-[18F]fluoro-D-glucose positron emission tomography uptake in a giant adrenal myelolipoma. AB - Incidental adrenal lesions found on anatomic imaging are not uncommon. 2-Deoxy-2 [18F]fluoro-D-glucose positron emission tomography (FDG-PET) imaging is highly accurate in the differentiation of benign from malignant adrenal lesions, both in patients with proven malignancy and with adrenal lesions detected incidentally. A 60-year-old white female with a history of lower mid-back pain underwent computerized tomography (CT) imaging that identified a 15-cm complex mass within the left adrenal gland with soft tissue, cystic, and adipose components. FDG-PET imaging showed significant hypermetabolic activity within portions of the mass with central photopenia suggesting a malignant lesion with central necrosis. Surgical excision and pathological examination, however, revealed a benign adrenal myelolipoma with extensive adenomatous and hematopoietic elements. Prior reports of adrenal myelolipoma evaluated with FDG-PET imaging have described no significant FDG uptake within these benign tumors. This case is an unusual example of histologically proven benign adrenal myelolipoma that was hypermetabolic on FDG-PET imaging. Correlation of pathologic and imaging findings demonstrated that the hypodense regions on CT were hypometabolic on FDG-PET and corresponded to cystic necrosis and adipose elements, whereas the adenomatous and hematopoietic elements were hypermetabolic. PMID- 14537111 TI - Cerebral lesions incidentally detected on 2-deoxy-2-[18F]fluoro-D-glucose positron emission tomography images of patients evaluated for body malignancies. AB - OBJECTIVES: The purpose of this work was done to evaluate the value of including the brain in the field of view of a whole-body 2-deoxy-2-[18F] fluoro-D-glucose positron emission tomography (FDG-PET) study of patients referred for the evaluation of body malignancies. METHODS: A total of 1026 consecutive patients were included in this work. The primary diagnoses were the following: lung (n = 253), colorectal (n = 148), head and neck (n = 61), lymphoma (n = 249), melanoma (n = 84), and others (n = 231). Whole-body FDG images including the brain were acquired with a dedicated PET tomograph (GE advance, General Electronic Medical Systems, Milwaukee, WI) one hour after the intravenous administration of 10 mCi of FDG. Two experienced nuclear medicine physicians interpreted the images. Positive findings in the brain or the skull were correlated with other imaging studies and clinical follow-up. RESULTS: Abnormal findings were detected in 3.9% (40/1026) of the patients. Among the 40 abnormal focal lesions, 29 patients had a known history of cerebral disease, cerebrovascular or metastatic disease in most patients. Of the 11 patients without a prior history of cerebral disease, four patients had increased focal FDG uptake suggestive of metastases. Among these, two were proven clinically, one was proven to be a skull base metastasis on MRI, and the other had negative clinical follow-up, but only of two months duration. The other seven patients had a decreased focal FDG uptake most consistent with infarct, one was proven clinically, and the other six had a negative clinical follow-up (mean of 6.3 months, range 1-10), but had multiple risk factors for cerebrovascular disease. CONCLUSIONS: We conclude that FDG-PET screening for cerebral lesions in patients with body malignancy has little clinical impact. Unsuspected cerebral or skull metastases were detected in 0.4% (4/1026) of the patients. PMID- 14537112 TI - High 2-deoxy-2-[18F]fluoro-D-glucose accumulation in a case of retroperitoneal fibrosis following resection of carcinoid tumor. AB - Positron emission tomography (PET) using 2-deoxy-2-[18F]fluoro-D-glucose (FDG) has been shown to be a highly sensitive diagnostic tool to stage, restage, and monitor the progress of various neoplasms. A number of physiological and non neoplastic conditions, however, also may be associated with focal accumulation of FDG and can cause false-positive results. This work reports a 52-year-old man who had marked FDG accumulation in carcinoid tumor of the distal ileum. The tumor was resected. A follow-up FDG-PET scan one and a half years later revealed intense FDG accumulation in the abdominal periaortic region, suggesting recurrent malignancy. Computerized tomography (CT) of the abdomen showed soft tissue mass surrounding the aorta. Fine-needle aspiration biopsy of the periaortic soft tissue confirmed benign retroperitoneal fibrosis. This case shows that retroperitoneal fibrosis can cause intense FDG accumulation giving false impression of malignancy. In interpretation of whole-body FDG-PET, various physiological and benign causes of FDG accumulation must be considered in order to avoid pitfalls. The authors have reviewed the literature and discussed association of carcinoid tumor and retroperitoneal fibrosis. PMID- 14537113 TI - Comparison of an 18F labeled derivative of vasoactive intestinal peptide and 2 deoxy-2-[18F]fluoro-D-glucose in nude mice bearing breast cancer xenografts. AB - PURPOSE: A 18fluorine-labeled derivative of vasoactive intestinal peptide [18F- Arg,Arg VIP(18F-dVIP)] was evaluated as a potential imaging agent for breast cancer by comparison with 2-deoxy-2-[18F]fluoro-D-glucose (FDG) using standard ex vivo determinations and small animal position emission tomography (PET) imaging. PROCEDURES: Human breast carcinomas, T-47D and MDA-MB231, tumor-bearing nude mice were injected intravenously with 18F-dVIP or FDG for imaging and/or biodistribution (ex vivo) determined by gamma counting. RESULTS: FDG had two- to three-fold greater tumor accumulation and target-to-non target contrast relative to 18F-dVIP. VIP receptors were detected in both tumor types but in low concentrations (<15,000 receptors/cell) consistent with lower uptakes. FDG was cleared rapidly from non-target tissues while 18F-dVIP cleared into the kidneys. CONCLUSIONS: 18F-dVIP uptake in mice T-47D tumors and kidneys determined by imaging correlated with values determined by ex vivo counting suggesting that tumor and other tissue uptakes can be quantified by in vivo positron projection imaging. PMID- 14537114 TI - Sub-millimeter technetium-99m calibration sources. AB - PURPOSE: Small animal radioscintigraphic imaging systems aim to achieve sub millimeter resolution. At the present time, sub-millimeter calibration sources that can be placed at will within an imaged volume are not readily available. We have developed a method for producing technetium-99m (Tc-99m) sources in less than 15 minutes with readily available reagents. PROCEDURES: Tc-99m pertechnetate [TcO(4)](-) was incubated with 45 microm to 106 microm diameter spherical anion exchange beads, washed, and mounted as desired for instrument calibration. RESULTS: The procedure yields spherical sources having between 6.8 microCi to 11.1 microCi of Tc-99m per source. This work shows that dual imaging of these sources using white light and radioscintigraphy permits measurement of system performance with high precision. CONCLUSION: Easily prepared, sub-millimeter Tc 99m spherical calibration sources are described, and it is demonstrated that such sources are useful for measuring the resolution and sensitivity of radioscintigraphic systems, such as those designed for small animal imaging. PMID- 14537115 TI - 11C-choline and 2-deoxy-2-[18F]fluoro-D-glucose in tumor imaging with positron emission tomography. AB - This review summarizes the result of the distribution studies of 11C-choline (CH) and 2-deoxy-2-[18F]fluoro-D-glucose (FDG) using positron emission tomography (PET) in many patients with various tumors. The procedures were as follows: CH PET, following transmission scan, emission scan started forty minutes after injection of CH: FDG-PET, following transmission scan, emission scan started forty minutes after injection of FDG. The final images were displayed on a scale of standardized uptake value (SUV). Various tumors were visualized with both CH and FDG. If normal organs took up radioactivity high, however, it was impossible to distinguish the tumor uptake from the normal organ uptake. The following organs showed high uptake: with CH, kidney, liver, pancreas, small intestine juice, and salivary gland; with FDG, brain, heart, and urine. If there was no interference with normal organ uptake, CH-PET visualized tumors as small as 5 mm in diameter, and FDG-PET visualized tumors of 10 mm in diameter. Lung cancer and pulmonary tuberculosis could be differentiated, in many cases, by comparing CH PET and FDG-PET. In conclusion, CH-PET is very useful for detecting various tumors, particularly if it was combined with FDG-PET. In addition, this review also discusses the rationale for the use of CH-PET for tumor imaging, the dosimetry of CH, and some 18F-substituted choline analogs. PMID- 14537116 TI - A patient with primary cardiac lymphoma. PMID- 14537117 TI - Inadvertent 2-deoxy-2-[18F]fluoro-D-glucose lymphoscintigraphy: a potential pitfall characterized by hybrid PET-CT. AB - PURPOSE: To communicate an important pitfall in positron emission tomography (PET) oncologic staging using 2-deoxy-2-[18F]fluoro-D-glucose (FDG). PROCEDURE: A 60-year-old man with a right upper lobe bronchogenic carcinoma was injected with FDG through a cannula in his left cubital fossa, with a small amount of the dose extravasating. RESULTS: Images obtained one hour post-injection with a positron emission tomography-computerized tomography (PET-CT) scanner demonstrated linear FDG activity running with the left arm brachial vascular bundle and along collateral channels, and accumulation of FDG in a low left axillary lymph node of normal size and CT appearance. CONCLUSIONS: Following inadvertent extravasation, FDG appears to accumulate in regional lymph nodes in the same fashion as bone scanning agents or lymphoscintigraphy colloid. Physicians and technologists need to be aware of this important pitfall in order to avoid the false positive diagnosis of nodal metastases. The ability of PET to demonstrate lymphatics and lymph nodes indicates its possible future use in lymphoscintigraphy. PMID- 14537118 TI - 2-Deoxy-2-[18F]fluoro-D-glucose positron emission tomography monitoring disease progress in a patient with islet carcinoma of the pancreas. AB - A woman with islet cell carcinoma of the pancreas proven by biopsy at an exploratory laparotomy underwent six cycles of chemotherapy with paclitaxel and carboplatin. After the chemotherapy to monitor her primary tumor and hepatic metastases, she had three consecutive 2-deoxy-2-[18F]fluoro-D-glucose positron emission tomography (FDG-PET) determinations that correlated with an In-111 octreotide single photon emission computer tomography. FDG-PET might have utility in monitoring islet cell carcinoma of the pancreas in evaluating metabolic changes following chemotherapy. PMID- 14537119 TI - Added clinical benefit of incorporating 2-deoxy-2-[18F]fluoro-D-glucose with positron emission tomography into the clinical evaluation of patients with cognitive impairment. AB - PURPOSE: Growing evidence indicates that appropriate incorporation of positron emission tomography (PET) into the evaluation of patients with early symptoms of cognitive decline can improve diagnostic and prognostic accuracy. In the present work, an explicitly defined role for PET and its associated impact on expected clinical outcomes were systematically examined. PROCEDURES: We compared the relative value of two strategies for assessing whether Alzheimer's disease (AD) was responsible for cognitive decline in geriatric patients, and in subsequently managing those patients according to the recommended standards of the American Academy of Neurology (AAN). The first strategy was based on an approach already endorsed by the AAN, following evidence-based reviews carried out by its quality standards subcommittee. The second approach was based on many of the same AAN recommendations-with respect to initial general medical and neurologic examination, structural imaging and laboratory tests, as well as ultimate management-but additionally incorporated PET in appropriate cases, to determine the presence or absence of a pattern of regional cerebral metabolism characteristic of AD. Clinical outcomes accruing to each strategy were calculated using formalized tools of decision analysis. RESULTS: The strategy making use of PET increased diagnostic accuracy, yielding decreased rates of both false negative (from 8.3 to 3.1%) and false positive (from 23.0 to 11.9%) diagnoses for AD, compared with the conventional strategy. When coupled with AAN treatment recommendations for patients having (or not having) non-severe AD, these differences in diagnostic accuracy corresponded to approximately a 62% decrease in avoidable months of nursing home care, and a 48% decrease in months of unnecessary drug therapy resulting from inaccurate diagnoses. The benefit in clinical outcome of the proposed strategy was maintained over a wide range of values for sensitivity, specificity, and projected impact on need for nursing home care. CONCLUSION: Use of PET for evaluating early cognitive decline in geriatric patients can add valuable information to the clinical assessment, resulting in a greater number of patients being accurately diagnosed and properly treated. PET can be used to diminish disease-related and treatment-related morbidity of dementia, through earlier institution of appropriate management. PMID- 14537120 TI - Standardized Uptake Values in 2-Deoxy-2-[18F]Fluoro-D-Glucose with Positron Emission Tomography. Clinical Significance of Iterative Reconstruction and Segmented Attenuation Compared with Conventional Filtered Back Projection and Measured Attenuation Correction. AB - PURPOSE: To evaluate the clinical significance of differences in 2-deoxy-2 [18F]fluoro-D-glucose positron emission tomography (FDG-PET) lymph node standardized uptake values (SUV) in human immunodeficiency virus (HIV) infection using iterative reconstruction with segmented attenuation correction (IR SAC) compared to filtered back-projection with measured attenuation correction (FBP MAC). PROCEDURES: Seven patients with HIV infection and multiple focal lymph node abnormalities were investigated with whole-body FDG-PET. Mean and maximum SUVs from lymph node regions of interest (n = 961) were compared for quantitative differences between reconstruction techniques. RESULTS: IR MAC resulted in significantly lower mean SUV [0.06; 95% (confidence interval (CI)) = 0.04-0.07] and maximum SUV (0.82; 95% CI = 0.77-0.88) values compared to FBP MAC. With IR, segmentation of attenuation correction (AC) resulted in significantly higher mean SUV (0.12; 95% CI = 0.11-0.13) and maximum SUV (0.21; 95% CI = 0.18-0.23) values compared to IR MAC. The overall effect of both IR and SAC was a slight but significant increase in mean SUV (0.06; 95% CI = 0.06-0.08; bias = 2.1%) and a significant decrease in maximum SUV (0.62; 95% CI = 0.56-0.67) compared to FBP MAC. CONCLUSIONS: With our reconstruction parameters, significant differences in mean and maximum SUV values were observed. The magnitude of the mean SUV difference, however, was small. IR SAC is a promising method to accurately quantify standardized uptake values for clinical use. PMID- 14537121 TI - Evaluation of response to neoadjuvant therapy by quantitative 2-deoxy-2 [18F]fluoro-D-glucose with positron emission tomography in patients with esophageal cancer. AB - PURPOSE: This work evaluates positron emission tomography (PET) with 2-deoxy-2 [18F]fluoro-D-glucose (FDG) in assessing response to therapy in patients with esophageal cancer. PROCEDURES: Twenty-four patients underwent FDG-PET before (pre Rx) and after (post-Rx) chemoradiation therapy; 20 then underwent esophagectomy. The response of the primary tumors was visually assessed, and tumor volume, peak tumor standardized uptake value (SUV(peak)), average SUV (SUV(ave)), and total lesion glycolysis were determined pre-treatment and post-treatment. Patients were divided into groups according to the absence (Group A) or presence (Group B) of residual tumor after neoadjuvant therapy. RESULTS: Among the quantitative PET parameters for Group A (n = 6) and Group B (n = 18), only change in tumor volume identified complete responders. Quantitative PET indices were not different in patients with or without post-Rx esophagitis. CONCLUSIONS: The change in tumor volume identifies patients with complete response to neoadjuvant therapy, and quantitative evaluation of the primary tumor cannot separate post-Rx inflammation from residual tumor. PMID- 14537123 TI - Characterization of uptake of 2-deoxy-2-[18F]fluoro-D-glucose by fungal associated inflammation: the differential uptake ratio for blastomyces-associated lesions is as high as for lymphoma and higher than for turpentine abscesses in experimentally induced lesions in rats. AB - PURPOSE: The objective of this investigation was to determine the biologic basis and the significance of uptake of 2-deoxy-2-[18F]fluoro-D-glucose (FDG) using experimentally created fungal lesions in rats. PROCEDURES: Uptake of FDG by experimentally induced Blastomyces granulomas was compared with uptake by turpentine abscesses (Group 1) and by lymphomas (Group 2) using the differential uptake ratio (DUR) measured one hour after administration of 2 mCi FDG intravenously. Frozen tissue sections of Blastomyces lesions and turpentine abscesses were placed in contact with radiographic film for macroautoradiography. RESULTS: In rats in Group 1, the median (range) DUR for the Blastomyces granulomas was 1.9 (1.1-2.6) and was significantly higher than the DUR for turpentine abscesses 0.9 (0.6-1.4) and muscle 0.2 (0.1-0.5; P < 0.001). In Group 2, the median (range) DUR for the Blastomyces granulomas, lymphomas, and muscle from the rats in Group 2 were 1.8 (1.2-3.4), 1.9 (1.0-4.0), and 0.2 (0.1-0.3), respectively. There was no significant difference between the DUR of Blastomyces granulomas and lymphomas. Macroautoradiographs of the Blastomyces granulomas revealed intense uptake of FDG in the region occupied by the yeast organisms and the granulomatous inflammation. CONCLUSIONS: Blastomyces granulomas typically have high uptake of FDG associated with the region composed of the granulomatous inflammatory reaction and Blastomyces yeast organisms. PMID- 14537124 TI - Characterization of uptake of 2-deoxy-2-[18F] fluoro-D-glucose by fungal associated inflammation: the standardized uptake value is greater for lesions of blastomycosis than for lymphoma in dogs with naturally occurring disease. AB - PURPOSE: Based on limited reports, fungal lesions can have remarkably high intensity uptake of 2-deoxy-2-[18F]fluoro-D-glucose (FDG) on positron emission tomography (PET) images. The purpose of this investigation was to compare the standardized uptake value (SUV) of naturally occurring lesions of blastomycosis with the SUV of naturally occurring lymphoma in a series of dogs. PROCEDURES: Five dogs with naturally occurring blastomycosis and three dogs with lymphoma underwent whole-body FDG-PET prior to receiving any treatment for their disease. RESULTS: The (mean +/- SD) SUV for 13 blastomycosis lesions was 7.7 +/- 2.0 versus a mean for 17 lymphomas of 4.8 +/- 1.8. These values were significantly different (P = 0.0537). There was overlap between the SUV of Blastomyces associated lesions versus lymphomas, but a cut-off SUV of 7.0 was 100% specific for Blastomyces lesions. Numerous sites of disease were detected on the FDG-PET images that were not detected clinically. CONCLUSIONS: FDG-PET is useful for determining the extent of disease in dogs with blastomycosis. The SUV for Blastomyces-associated lesions are as high or higher than for malignant lymphoma. Due to the similarities in canine and human blastomycosis and lymphomas, similar results would be predicted in human patients. In regions where blastomycosis is endemic, Blastomyces granulomas should be considered a differential diagnosis for lesions with high intensity uptake of FDG. PMID- 14537125 TI - Imaging and therapeutics: the role of neuronal transport in the regional specificity of L-DOPA accumulation in brain. AB - PURPOSE: To investigate the in vitro regional accumulation of L-3,4 dihydroxyphenylalanine (L-DOPA) in brain tissue. PROCEDURE: Neuronal membrane transport of L-DOPA was investigated in rat and squirrel monkey brain tissue. The kinetics of L-DOPA regional transport was characterized, and the effect of amino acids on transport was evaluated using isolated nerve terminals from striatum and cerebral cortex. RESULTS: When L-DOPA uptake was measured in modified Krebs Ringer medium, transport occurred in both synaptosome preparations. In the presence of dilute protein-free plasma, uptake of L-DOPA was significantly present in striatal nerve terminals (P < 0.005), but was completely inhibited in terminals isolated from the cortex. L-DOPA transport in striatal synaptosomes was primarily inhibited by large neutral aromatic L-amino acids, in contrast to that in cortical synaptosomes that was mainly affected by large neutral aliphatic L amino acids. A saturable component of influx was detected in both synaptosome preparations, where kinetic analysis revealed that the relative affinity of L DOPA was greater for the carrier in the striatum than in the cortex. Based on the distribution of neuronal cell types within the two regions and the effect of 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) lesioning in squirrel monkeys, the striatal specific accumulation of L-DOPA likely occurs within dopaminergic terminals. CONCLUSIONS: These results demonstrate that the in vivo regional specificity of L-DOPA localization in brain tissue is primarily controlled by neuronal transport. PMID- 14537126 TI - Iterative image reconstruction for clinical PET using ordered subsets, median root prior, and a web-based interface. AB - PURPOSE: The development, implementation and validation of simple, flexible and efficient iterative image reconstruction (IIR) methods for their take-up in routine clinical positron emission tomography (PET) static or dynamic studies. PROCEDURES: The ordered subsets (OS) technique applied for the acceleration of the maximum likelihood expectation maximization (MLEM) IIR algorithm is here extended to include the weighted least-squares (WLS), image space reconstruction algorithm (ISRA) and the space alternating generalized EM (SAGE). The median root prior (MRP) has been successfully applied as a Bayesian regularization to control the noise level in the reconstructed images. All methods are implemented on distributed Pentium systems and tested using simulated PET data from a brain phantom. A Javascript is used for the initiation of the reconstruction. RESULTS: Taking into consideration the image quality and the time required for the reconstruction, the MRP-OSEM (ordered subsets expectation maximization) seems to provide best results after four to eight iterations, with four subsets and a MRP coefficient of 0.2-0.4. Iterative reconstruction of the transmission images with OS-acceleration and MRP regularization with subsequent calculation of the attenuation correction factors (ACFs) is shown to effectively remove streak artifacts in the emission images, especially along paths of high attenuation. CONCLUSIONS: An efficient implementation using distributed processing principles and a web-based interface allows the reconstruction of one frame (with 63 cross section slices) from a dynamic determination in few minutes. This work showed that regular PC systems can provide fast execution and produce results in clinically meaningful times. This eradicates the argument of the computational burden of the method that prevented the extensive use of IIR in today's modern PET systems. PMID- 14537127 TI - Reduction of myocardial 2-deoxy-2-[18F]fluoro-D-glucose uptake artifacts in positron emission tomography using dietary carbohydrate restriction. AB - PURPOSE: The use of carbohydrate restriction prior to 2-deoxy-2-[18F] positron emission tomography (FDG-PET) to reduce image artifacts caused by myocardial FDG uptake was investigated. PROCEDURES: Ninety-six whole-body FDG-PET scans were studied. Dietary intake of the meal prior to scanning was recorded. Coronal and transverse images were assessed visually and scored based on myocardial FDG uptake and the presence of associated image artifacts. RESULTS: Of 49 patients who consumed carbohydrates prior to PET, 14 (28.6%) had a clinically significant image artifact versus only five (10.6%) of 47 patients who did not consume carbohydrates (P = 0.0275). A 3.36 times increased risk (confidence interval 1.00 12.97) for clinically significant image artifacts was calculated for patients who consume carbohydrates in their last meal prior to scanning. CONCLUSIONS: With the rising use of FDG-PET in the evaluation of intra-thoracic malignancies, dietary carbohydrate restriction prior to scanning may play a role in increasing lesion detectability and reducing false negative scans. PMID- 14537128 TI - Optimizing imaging time for improved performance in oncology PET studies. AB - PURPOSE: The potential for improving the diagnostic performance of static positron imaging tomography (PET) by judiciously choosing optimum post-injection imaging times is investigated. PROCEDURES: Dynamic and whole-body scan data, from 2-deoxy-2-[18F]fluoro-D-glucose (FDG) oncological studies, are analyzed for changing standardized uptake value (SUV) behavior with increasing post-injection times at either single- or multiple-bed positions. Model-based interpretations address d(SUV)/dt, shown to correlate with SUV, and the contrast ratio for a tumor and its surroundings. A method for correcting measurements to a standardized time is given. RESULTS: Both data and model-based equations suggest that starting data acquisition later than the average 55 +/- 15 (SD) minutes post injection reported in the FDG literature can improve contrast ratios. Considerations for choosing an optimum time from a clinical standpoint are listed. CONCLUSIONS: It is concluded that the appropriate time for each particular protocol can be found with the aid of the information presented here. True optimization, however, remains a complex issue. PMID- 14537129 TI - The use of 2-deoxy-2-[18F]fluoro-D-glucose (FDG-PET) positron emission tomography in the routine diagnosis of epilepsy. AB - PURPOSE: Positron emission tomography with 2-deoxy fluoroglucose positron emission tomography (18-FDG-PET) is widely used in the pre-surgical evaluation of subjects with epilepsy, but little is known of its usefulness in a non-surgical population. PROCEDURES: We analyzed the sensitivity of PET as a diagnostic tool in a large unselected population of epilepsy subjects. Pre-surgical and non surgical portions of this population were individually assessed as well. The relationship of PET abnormalities to other neurodiagnostic tests was examined. Statistical assessment relied primarily on contingency tables (chi-square tests), with ANOVA or non-parametric assessment used as necessary. RESULTS: While PET was more likely to identify areas of decreased metabolism in the surgical population than in the non-surgical populations, it nevertheless found a significant number of abnormalities in the total population and in the non-surgical group alone. Even in groups in which the clinical diagnosis was unknown, abnormalities were found 40% of the time. PET was useful as an exclusionary diagnostic tool for non epileptic seizures (NES) and primary generalized epilepsies (PGE) with sensitivity, specificity, and accuracy > 90%. The PET was somewhat more sensitive than magnetic resonance imaging (MRI) in finding abnormalities in the total population, but was less sensitive than electroencephalography (EEG). CONCLUSION: PET may be a useful diagnostic tool in the general epilepsy population even when a definitive clinical diagnosis is not suggested by other modalities. PMID- 14537130 TI - Quantitative measurement of regional blood flow in hepatic focal nodular hyperplasia. Findings on positron emission tomography. AB - The blood flow through normal liver parenchyma and a pathologically proven hepatic region of focal nodular hyperplasia (FNH) of the lateral lobe, was measured in a single patient in a quantitative manner using regional dynamic positron emission tomography (PET) imaging with O-15 carbon dioxide (steady state) and O-15 water (dynamic state). The steady state PET images revealed higher radioactivity concentration in the lateral lobe than in the surrounding normal liver parenchyma. Regional and total blood flow in the region of FNH, as determined by the dynamic state method, was 292.2 ml/100g/min and 330.2 ml/min, respectively. It was determined that the region of FNH in this patient had higher blood flow than that described previously for liver tumors such as hepatocellularcarcinoma and colorectal hepatic metastases. PMID- 14537131 TI - 2-Deoxy-2-[18F]fluoro-D-glucose positron emission tomography in target volume definition for radiotherapy of patients with non-small-cell lung cancer. AB - PURPOSE: To discuss the potential contribution of 2-deoxy-2-[18F]fluoro-D-glucose positron emission tomography (FDG-PET) in radiotherapy planning for non-small cell lung cancer (NSCLC) considering new concepts on target volume definition. PROCEDURES: Recent investigations on the topic are reviewed with regard to current concepts of target volume definition for NSCLC. RESULTS: As intrathoracic recurrence is the leading cause of death after primary radiotherapy of NSCLC, there is a need for improving local control by escalating treatment intensity to gross disease. The value of elective nodal irradiation (ENI), resembling prophylactic irradiation of macroscopically unaffected parts of the mediastinum, is being considered. CONCLUSION: As FDG-PET has been shown to enhance the diagnostic accuracy of computed tomography (CT), and to have a potentially high impact on the identification of malignant tissue, it should be implicated in prospective clinical trials on dose escalation and three-dimensional conformal radiotherapy, especially in those including a reduction of target volumes. PMID- 14537134 TI - The planning and design of a new PET radiochemistry facility. AB - The objectives of the Mayo positron emission tomography (PET) radiochemistry facility are the production of PET drugs for clinical service of our in-house patients, commercial distribution of PET drug products, and development of new PET drugs. The factors foremost in the planning and design phases were the current regulatory climate for PET drug production, radiation safety issues, and effective production flow. A medium-energy cyclotron was preferred for its small footprint to allow a compact vault, its high-proton energy to offer a higher product radioactivity; and its research capabilities. A vault installation was chosen instead of a self-shielded machine for improved access and ease of maintenance. Adjacent to the cyclotron is an area that houses the support equipment and a large dedicated workshop to support machine maintenance and targetry development. The total floor area of the PET radiochemistry facility is 344.2 m(2) (3,705.5 ft(2)), of which the radiochemistry laboratory occupies 130.7 m(2) (1,407 ft(2)). To reduce environmental contamination of PET drug products, the laboratory contains a controlled-air environment class 10,000 (M5.5) clean room with access via an interlocking entry change area. A fully shielded isolator (class 100 [M3.5]) is located in the clean room. The PET drugs are delivered via shielded tubing between the synthesizer and isolator. Inside the isolator, there is an automated device for dispensing the PET drug into either a bulk-activity vial or a unit-dose syringe. The dispensed PET radiopharmaceutical then passes through a hatch to a dedicated area where it is packaged for in-house use or commercial distribution. Unit doses for in-house patients are transported via pneumatic tube to the PET imaging area 76.2 m (250 ft) away. There is extensive radiation area monitoring throughout the facility that continuously measures radiation levels. We believe that our new PET radiochemistry facility not only meets overall objectives, but also provides an ergonomic, efficient working environment for the production and development of PET drugs. PMID- 14537136 TI - Cerebral blood flow during propofol induced sedation. AB - PURPOSE: This work determined if 2, 6-diisopropylphenol (propofol) selectively affects cerebral blood flow in regions associated with wakefulness. PROCEDURES: Cerebral blood flow (CBF) was measured with positron emission tomography (PET) using the 15O-water bolus technique in 10 subjects while awake and during light and deep sedation. Arterial blood was sampled for CBF estimation, blood gases and propofol plasma concentrations. RESULTS: Global CBF decreased under deep sedation. A regression analysis of CBF vs. propofol concentration showed significant decreases in CBF in the thalamus and posterior cingulate and increases in the hippocampus and cerebellum. An ANCOVA analysis on condition (controlling for pCO(2) levels) showed mean CBF decreased in the thalamus and posterior cingulate cortex and increased in the primary motor and hippocampal areas during the light and deep sedation compared to awake conditions. CONCLUSIONS: These data support the hypothesis that propofol preferentially alters CBF in specific brain regions necessary to maintain wakefulness. PMID- 14537137 TI - A comparative study on the uptake and incorporation of radiolabeled methionine, choline and fluorodeoxyglucose in human astrocytoma. AB - PURPOSE: The goal of this investigation was to evaluate uptake and incorporation of 2-deoxy-2-[18F]fluoro-D-glucose (FDG), 11C-methionine, and 11C-choline in 17 patients suspected of grade-II and grade-III tumors using positron emission tomography (PET) and use in vitro astrocytoma cell lines in order to support in vivo findings. METHODS: Seventeen patients with suspected astrocytomas (9 grade II and 8 grade-III) were studied by PET with FDG and 11C-methionine; and one patient (grade-III) with FDG, 11C-methionine and 11C-choline. Uptake of PET molecular imaging probe was quantitative based on tumor to corresponding contralateral-region uptake ratio, tumor to mean-cortical-uptake ratio, and tumor to white matter uptake ratio. This was correlated with World Health Organization histology grading system and clinical follow-up. Uptake and incorporation of 3H methionine, 3H-choline and FDG into lipid, RNA, DNA, and protein were investigated in a grade-III human tumor brain-14 astrocytoma cell line. RESULTS: A time-dependent increase in the total uptake of 3H-methionine, 3H-choline and FDG was observed in human tumor brain-14 astrocytoma-III cell line. 3H-methionine was incorporated predominantly into proteins (in excess of 40% at 1 h) while 3H choline incorporated primarily into lipids (in excess of 60% at 1 hr). Total uptake of FDG was accounted for in the free-pool supernatant fraction. In all patients, PET images of 11C-methionine and FDG provided higher tumor to white matter ratios than tumor to corresponding contra-lateral region ratios and tumor to mean cortical uptake ratios. In grade II patients, FDG did not exhibit significant increase in tumor uptake, while 11C-methionine was a good predictor with ratios of approximately 1.50 +/- 0.48. In grade III patients, both FDG and 11C-methionine exhibited higher ratios than for grade II, with 11C-methionine being the greatest (ratios of 2.50 +/- 0.85), possibly suggesting enhanced protein synthesis. With respect to tumor delineating potential, 11C-choline may be equal to or slightly better than 11C-methionine in the subject evaluated with all three probes. CONCLUSIONS: Results suggest that a combination of FDG and 11C methionine is useful in the prediction of histological grade of astrocytomas. In addition, 11C-methionine is better than FDG in delineating tumor boundary for low grade gliomas. In vitro results suggest that 3H-methionine is significantly incorporated into proteins and provides the major driving force in the uptake of 11C-methionine observed in PET images. PMID- 14537138 TI - The receiver operating characteristic curve for the standard uptake value in a group of patients with bone marrow metastasis. AB - PURPOSE: The aim of this work was to determine the standard uptake value (SUV) threshold for differentiating malignant from benign bone lesions. MATERIAL AND METHODS: Ninety-nine bone sites in 33 patients who had undergone a 2-deoxy-2 [18F]fluoro-D-glucose-positron emission tomography (FDG-PET) study for cancer evaluation were studied. In addition to FDG-PET, a bone scan and at least two of the following determinations: magnetic resonance imaging (MRI), computed tomography (CT), and x-ray were conducted in each patient. The bone lesions were considered positive for malignancy if confirmed by clinical follow-up or a high degree of suspicion based on the positive results of at least three (which must include bone scan) out of four other imaging modalities. By these criteria, 39 lesions were considered positive and 60 were considered negative. The SUV values were classified as positive or negative using 61 different values of threshold (range from 1.0 to 7.0). These results were compared with the positive criteria above and reclassified as true positive, true negative, false positive, and false negative. The true-positive fraction and false-positive fraction were calculated for each threshold value. The receiver operating characteristic (ROC) curve was drawn and the best value was determined by visual analysis. RESULTS: The SUV threshold was considered 2.5. Twenty-nine out of 39 bone lesions classified as positive showed a SUV > 2.5. Of the 10 false-negative lesions, seven showed a SUV between 1.1 and 2.0, and three were not detected. Fifty-six out of 60 lesions classified as negative showed a SUV < 2.5. Four lesions were false positive: one was a rib fracture and three were severe degenerative changes in the lumbar spine. Using an SUV threshold of 2.5, the sensitivity was 74.3% and the specificity was 93.3%. CONCLUSION: In our patient population, the optimal SUV to classify a bone lesion as malignant or benign is 2.5. PMID- 14537135 TI - [15O]water pharmacokinetics: influence of age and gender in normal subjects. AB - PURPOSE: To compile a normal database for the characterization of global [15O]water pharmacokinetic behavior. PROCEDURES: The influences of age, gender, and body habitus on the pharmacokinetics of [15O]water were investigated in a series of normal subjects, N = 100 (50 males, 50 females, age = 19-79) who were participants in cognitive activation studies. Arterial blood was analyzed by autosampler and parametric images were constructed using a 40-second summed image and the autoradiographic model. RESULTS: Males and females were comparable with respect to age, number of injections administered, and dose (mCi) administered per injection but differed significantly with respect to height, weight, and normalized dose (mCi/kg). There were significant gender-based differences in the bolus arrival time, global cerebral blood flow (gCBF), area-under-the-curve (AUC), summed image concentration, and dose-normalized concentration but not dose normalized AUC. Bolus arrival time, gCBF and dose-normalized AUC were significantly influenced by age. CONCLUSION: Age and gender are significant determinants of [15O]water pharmacokinetic behavior. PMID- 14537139 TI - Comparison of attenuation-corrected and non-corrected FDG-PET images for axillary nodal staging in newly diagnosed breast cancer. AB - PURPOSE: The aim of this work is to compare the accuracy of non-attenuation corrected (NAC) and attenuation-corrected (AC) PET images using 2-deoxy-2 [18F]fluoro-D-glucose (FDG) in assessment of the axilla in patients with newly diagnosed, untreated, primary breast cancers, and to determine the frequency of extra-axillary findings. PATIENTS AND METHODS: FDG-PET was performed in 36 patients with breast cancer one hour following the intravenous injection of approximately 370 MBq of FDG. Patients were imaged prior to axillary dissection to prospectively confirm the presence or absence of axillary metastases. NAC and AC images were separately and independently reviewed in a blinded fashion by two readers. Imaging results were compared with final diagnoses obtained by surgery and pathology. RESULTS: Ninety-six positive axillary lymph nodes in 15 patients were histopathologically confirmed by surgery. The average areas under the ROC curve for NAC and AC image were 0.682 and 0.721, respectively. In patient-based analysis, the sensitivity in interpreting NAC images for the presence of metastases was low (53.3%) and comparable to that of AC images (46.7%). The specificity of NAC image was high (85.7%), but tended to be lower than that in AC (95.2%). The overall diagnostic accuracy for detecting axillary involvement in AC images (75.0%) was comparable to that of NAC images (72.2%). Extra-axillary disease was found in three patients. CONCLUSION: NAC images were comparable to AC images, although there were more false positive results with the NAC images. While AC PET has high specificity in this application, it appears insufficiently sensitive to use these methods to avoid axillary tissue sampling, when negative. Extra-axillary findings, while infrequent, may be quite important. Examination of both NAC and AC images is advised when assessing possible metastatic breast cancer to the axilla using PET. PMID- 14537140 TI - Reproducibility of common semi-quantitative parameters for evaluating lung cancer glucose metabolism with positron emission tomography using 2-deoxy-2-[18F]fluoro D-glucose. AB - PURPOSE: Positron emission tomography (PET) with 2-deoxy-2-[18F]fluoro-D-glucose (FDG) has been used for various cancers, but reproducibility of common utilized semi-quantitative parameters, such as the maximal single pixel standardized uptake value (SUV) and effective glycolytic volume (EGV), remains unknown. Knowledge of precision is essential for applying these parameters to treatment monitoring. The purpose of this investigation was to assess the precision of PET results obtained by repeated examinations of patients with untreated lung cancer. PATIENTS AND METHODS: Ten patients with lung cancer underwent two PET examinations within a week with no intervening treatment. The reproducibility of three parameters:((1) maximal SUV of 1 x 1 pixel anywhere in the tumor, calculated on the basis of predicted lean body mass [SULmax]; (2) highest average SUV at 4 x 4 pixels in the tumor adjusted by predicted lean body mass [SULmean]; and (3) EGV calculated by multiplying SUL by tumor volume), using PET images obtained at 50-60 min post-injection, were examined. Plasma glucose, insulin and free fatty acid levels were also monitored. RESULTS: The SULmax, SULmean, and EGV were measured with a mean +/- S.D. difference of 11.3% +/- 8.0, 10.1% +/- 8.2, and 10.1% +/- 8.0%, respectively. By multiplying SUL by plasma glucose concentration, the mean differences were slightly reduced to 7.2% +/- 5.8, 6.7% +/- 6.2, and 9.5% +/- 8.2, respectively. CONCLUSION: These data indicate that commonly used semi-quantitative indices of glucose metabolism on PET show high reproducibly. This supports their use in sequential quantitative analysis in PET, such as in treatment response monitoring. PMID- 14537141 TI - Detection of extramedullary infiltrates in acute myelogenous leukemia with whole body positron emission tomography and 2-deoxy-2-[18F]-fluoro-D-glucose. AB - This work reports on a female patient with acute myelogenous leukemia (AML) FAB M 5a with initial extramedullary leukemia (EML) in skin, breast, and synovia. A year after diagnosis she developed a histologically proven isolated recurrence of the EML in the right upper ankle. The bone marrow was still in complete remission. Conventional x-ray, magnetic resonance imaging (MRI), bone scintigraphy, and 2-deoxy-2-[18F]-fluoro-D-glucose whole-body positron emission tomography (FDG-PET) were performed. All images showed alterations in the lower leg. Shortly after, an isolated relapse of the AML was diagnosed in the right elbow. FDG-PET demonstrated this lesion as well as an unknown lesion in the subcutis due to EML. In the course of her illness, the patient underwent one more PET examination for therapy control. The present observations suggest that whole body FDG-PET may be valuable for the detection of EML and for the assessment of chemotherapeutic effects on identified lesions. PMID- 14537142 TI - The impact of 2-deoxy-2[18F] fluoro-D-glucose whole body positron emission tomography for managing patients with melanoma: the referring physician's perspective. AB - PURPOSE: Whole body positron emission tomography (PET) imaging with 2-deoxy 2[18F]fluoro-D-glucose (FDG) has been used successfully to diagnose and stage melanoma. The impact of FDG-PET, however, on patient stage and management from the referring physicians' perspective is unknown. PROCEDURES: A questionnaire was sent to referring physicians to investigate whether and how PET altered clinical decision in treatment of melanoma patients. Surveys were sent to referring physicians of every melanoma patient who had a PET scan performed at UCLA or the Northern California PET Imaging Center (NCPIC). Data were used to evaluate the impact of FDG-PET on clinical management of melanoma patients based on pre-PET and post-PET staging. Management changes were classified as inter-modality if therapy changed from one modality to another or intra-modality if changes were made within a treatment modality. RESULTS: Fifty-one questionnaires (response rate of 35%) have been received to date. Referring physicians indicated that whole body FDG-PET changed the clinical stage in 15 out of 51 (29%) patients: 10 (20%) were up-staged and five (10%) were down-staged. The PET findings resulted in inter-modality management changes in 15 out of 51 patients (29%). Intra modality management change occurred in nine patients (18%). CONCLUSION: From the referring physicians' perspective, FDG-PET has a major impact and results in management changes in 53% of patients with melanoma. PMID- 14537143 TI - Biosensors for environmental monitoring. AB - Increasing environmental legislation which controls the release and the levels of certain chemicals in the environment has created a need for reliable monitoring of these substances in air, soil and especially water. Conventional analytical techniques, although highly precise, suffer from the disadvantages of high cost, the need for trained personnel and the fact that they are mostly laboratory bound. Biosensors because of their specificity, fast response times, low cost, portability, ease of use and a continuous real time signal, can present distinct advantages in certain cases. Their biological base makes them ideal for toxicological measurements which are suited for health and safety applications. Over the last 3-4 years there has been an increase in the number of publications concerning biosensors for environmental monitoring, especially in the field of pesticide measurements. This paper reviews some of the more important developments over the past 3-4 years. PMID- 14537144 TI - Cell growth patterns in immobilization matrices. AB - Within an immobilized cell matrix, mass transfer limitations on substrate delivery or product removal can often lead to a wide range of local chemical environments. As immobilized living cell populations actively grow and adapt to their surroundings, these mass transfer effects often lead to strong, time dependent spatial variations in substrate concentration and biomass densities and growth rates. This review focuses on the methods that have been devised, both experimentally and theoretically, to study the non-uniform growth patterns that arise in the mass transfer limited environment of an immobilization matrix, with particular attention being paid to cell growth in polysaccharide gels. PMID- 14537145 TI - Applications of monoclonal antibodies in aquaculture. AB - Monoclonal antibodies (MAbs) currently are being applied to the study of fish immunology and fish infectious diseases. MAbs to fish immunoglobulins (Igs) have helped isolate fish Igs, identify heavy and light chain variants in fish Ig, study the ontogeny of B lymphocytes, and improve techniques for the measurement of fish Ig and specific antibodies (Abs). MAbs have been obtained against several leucocyte surface antigens and are being used as markers for different subsets of fish leukocytes: neutrophils, non-specific cytotoxic cells and cells responsible for the mixed leucocyte reaction. The sensitivity and specificity of many immunoassays for identifying fish pathogens have been improved by the use of MAbs. Further improvement of these tests is currently being attempted by the use of MAbs together with the polymerase chain reaction (PCR). Epizootiological studies of fish diseases are beginning to emerge from the use of these reagents and techniques. In the near future these new methods should detect low levels of pathogens in adult carriers and perhaps in eggs, thus helping to prevent the dissemination of fish diseases. MAbs to fish pathogens are also being used for passive immunization studies as well as for conformational probes in the development of genetically engineered vaccines. PMID- 14537146 TI - The use of immunological methods to detect and identify bacteria in the environment. AB - Immunological detection methods have become increasingly important in microbial ecology for the tracking of specific microorganisms and for community analysis. For a reliable application of these techniques, the monoclonal antibodies or polyclonal antisera used have to fulfill several quality criteria. Cross reactivity, cellular localization of the antigenic determinant, affinity characteristics and the expression of the antigenic determinant at environmental conditions have to be determined. Immunological methods can be used for the identification, quantification and enrichment of specific bacteria in extracts as well as for the visualization of cells in situ. The sensitivity of advanced immunological methods can be compared to PCR techniques. Using image processing of epifluorescence micrographs or confocal laser scanning microscopy, the immunofluorescence approach can now be applied to study complex environmental samples. PMID- 14537155 TI - Tissue-type plasminogen activator: characteristics, applications and production technology. AB - Plasminogen activators have immense clinical significance as thrombolytic agents for management of stroke and myocardial infarction. Tissue-type plasminogen activator (tPA) is generally preferred as being effective and safer than either urokinase or streptokinase type activators. Large-scale production of tPA became possible through groundbreaking developments in cell lines and bioprocess technology. Nevertheless, at thousands of dollars per treatment, tPA remains expensive. Enhancing cellular productivity and downstream product recovery through new approaches continue to be major challenges as discussed in this review. Recent clinical experience suggests the need for yet better fibrinolytic agents and attempts are underway to modify the tPA molecule to second generation products. Emerging trends in this field are outlined. PMID- 14537156 TI - Antibody production in plants. AB - Production of heterologous proteins in plants has become increasingly efficient due to recent advances in plant biotechnology. Heterologous proteins that have specifically attracted a great deal of attention are plant-produced monoclonal antibodies. A variety of applications for these so-called plantibodies have been explored since they were first expressed in tobacco seven years ago. Both full length antibodies and antibody fragments produced in transgenic plants offer many intriguing possibilities to plant molecular biologists and plant breeders. However, questions such as how cellular targeting influences the expression and accumulation of these proteins in plants still need to be answered before the technology can be used commercially, on a large-scale. PMID- 14537605 TI - Long circulating liposomes: past, present and future. PMID- 14537604 TI - Cryopreservation of mammalian embryos and oocytes: recent advances. AB - The cryopreservation of embryos of most domestic species has become a routine procedure in embryo transfer, and recently, advances have been made in the cold storage of mammalian oocytes. The ability to sustain viable oocytes and embryos from mammalian species at low temperature for prolonged periods of time has important implications to basic and applied biotechnology. Recent advances in the study of physico-chemical behaviour of different cryoprotectants, use of various macromolecule additives in cryoprotective solutions and isolation and use of proteins of plant and animal origin with antifreeze activity offers many new options for cryopreservation of oocytes and embryos of animal and human origin. At the same time rapidly developing methods of oocyte/embryo manipulation such as in vitro embryo production, embryo splitting, embryo biopsying for gene and sex determination, embryo cloning and the isolation of individual blastomers, create new challenges in cryopreservation. Very recent advances in the cryopreservation of mammalian oocytes, in vivo- and in vitro-derived embryos, and micromanipulated embryos are reviewed in this manuscript. PMID- 14537819 TI - Liquid emulsion membranes: principles, problems and applications in fermentation processes. AB - Liquid emulsion membranes (LEMs) have developed into a versatile technique for a variety of applications involving selective and controlled transport of biochemicals. Biological applications cover the controlled delivery of drugs from capsules, detoxification of the circulatory system, recovery of useful compounds from waste streams and selective separation of products from fermentation broths. This review traces the development of LEMs, discusses their key features, advantages and limitations, describes methods of modelling LEM systems and highlights some applications with industrial potential. Two kinds of LEM systems are considered. The first type are agitated emulsions, which are relatively easy to prepare and use but may be limited in their selectivity and long-term stability. Supported liquid membranes (SLMs) are a recent development; they use porous solid supports and have excellent stability and selectivity. Their chemical engineering aspects and applications in fermentation processes are considered. PMID- 14537820 TI - Fungal production of citric acid. AB - Citric acid is the principal organic acid found in citrus fruits. To meet increasing demands it is produced from carbohydrate feedstock by fermentation with the fungus Aspergillus niger and the yeasts of Candida spp. Effect of various fermentation conditions and the biochemistry of citric acid formation by A. niger have been discussed. Commercially citric acid is produced by surface, submerged and solid state fermentation techniques. Recovery of pure acid from fermentation broth is done primarily by precipitation with lime and also by solvent extraction. PMID- 14537821 TI - Prospects for the genetic manipulation of dairy cattle: opportunities beyond BST. AB - The dairy industry, with regulatory approvals of recombinant chymosin and bovine somatotropin (BST), has been at the forefront of food and agricultural biotechnology. The commercial fate of these products is one of several factors that may affect the success of future genetic manipulations in dairy cattle and dairy products. Other factors include technical and reproductive constraints in cattle and the cost of producing transgenic cattle. Early applications of genetic manipulation in cattle, for reasons of cost recoupment, may favor production of heterologous proteins in milk for pharmaceutical and medical use. Such applications could benefit genetic modification of milk and milk proteins for food use by providing advance knowledge and experience in mammalian protein expression. Other research opportunity areas that could affect prospects for genetic manipulation of dairy cattle include genome mapping, metabolic pathways, growth and development, and cattle/microbe interactions. PMID- 14537822 TI - Metabolic load and heterologous gene expression. AB - The expression of a foreign protein(s) in a recombinant host cell or organism often utilizes a significant amount of the host cell's resources, removing those resources away from host cell metabolism and placing a metabolic load (metabolic drain, metabolic burden) on the host. As a consequence of the imposed metabolic load, the biochemistry and physiology of the host may be dramatically altered. The numerous physiological changes that may occur often lowers the amount of the target foreign protein that is produced and eventually recovered from the recombinant organism. In this review the physiological changes to host cells, the causes of the phenomenon of metabolic load, and several strategies to avoid some of the problems associated with metabolic load are elaborated and discussed. PMID- 14538044 TI - Assessing brain biochemistry in the early evaluation of dementia: can we afford the improved diagnostic accuracy obtained by incorporating cerebral metabolic data acquired with positron emission tomography into the clinical work-up? PMID- 14538045 TI - 1999 ICP Distinguished Scientist Award. The history of positron emission tomography. AB - The history of Positron Emission Tomography (PET) is rich in technological achievements and advancements. The advancements that have benchmarked PET progress are the result of key components that include human intellect and passion for PET technology, relentless persuasion of key political forces to eliminate the barriers precluding PET usage, tireless efforts to raise awareness about PET and a crucial network of support throughout the PET community. This article sets forth a timeline of significant events that have contributed to the development of PET as it is known today. It introduces the earliest physicist and physician, for instance, who were responsible for the first medical applications for positron emitting radioisotopes using a simple brain probe that utilized coincidence to localize brain tumors. Additionally, it identifies landmark technological achievements that have helped pave the way to modern PET. This study includes historical accounts surrounding the use of the first human PET tomograph, discovery of the Bismuth Germanate (BGO) scintillator, development of the Fluorodeoxyglucose (FDG) PET method, the design of the first PET medical cyclotron with automated chemistry and operated by a PC and a technologist, Food and Drug Administration's approval of FDG, HCFA reimbursement, and the capacity of Lutetium Oxyorthosilicate (LSO) to produce a revolutionary advance in PET scanners. The main thrust of this article is to recognize via a timeline of PET accomplishments the noteworthy work of scientists, physicians and others who have been key players in various aspects of the continuous activity to move PET technology forward from invention to research, and to become a major clinical imaging modality. PMID- 14538046 TI - Tracing transgene expression in cancer gene therapy: a requirement for rational progress in the field. AB - This review summarizes the status of gene therapy in medicine and the role of molecular imaging in its development. In gene therapy, genetic material is introduced into cells in order to generate a specific biological effect. Natural (viruses) or artificial molecular constructs, named gene therapy vectors, are used to achieve efficient cell transduction. This new form of therapy can be used for treating a broad variety of conditions including hereditary diseases, infections, degenerative disorders and cancer. Monitoring transgene expression using noninvasive imaging techniques is a necessary complement for the development of clinical gene therapy. Recent developments in magnetic resonance imaging afford the possibility of detecting gene transfer in vivo, but the most promising results have been obtained with positron emission tomography (PET). PET allows imaging gene therapy products by administration of a labeled substrate when the transgene codes for an enzyme or by administration of a labeled ligand when the transgene codes for a receptor. In the latter strategy, a membrane molecule (somatostatin or dopamine receptors) is used to detect the selective trapping of its radiolabeled ligand in the transduced cells. One of the approaches for the genetic treatment of cancer consists in transferring the "suicide genes" into tumor cells, the most common being the thymidine kinase (tk) of herpes viruses. Different nucleoside analogs can be labeled for its use as PET reporter probes in order to visualize tk expression. The results of pre-clinical studies are extremely encouraging. Reliable methods for the in vivo tracing of transgene expression in humans have to be developed in order for the field of gene therapy to mature. PET has emerged as a powerful tool to assist in achieving this goal. PMID- 14538047 TI - FDG-PET in breast cancer: a different view of its clinical usefulness. AB - The current review addresses the role of whole-body PET imaging for diagnosing, staging, and re-staging of breast cancer, as well as its role for monitoring effects of treatment on breast cancer. The strengths and limitations of the conventional imaging modalities and those of PET are discussed. New solutions for relatively inexpensive PET screening of patients at risk for breast cancer are proposed. Clinical indications for PET in breast cancer are specified. PMID- 14538048 TI - PET scanners dedicated to molecular imaging of small animal models. AB - The dramatic advances of biological research in recent years that have focused on the molecular basis of how systems of the body (e.g. cells, organs and the whole organism) function, have increased the need for molecular imaging instrumentation. Of the several imaging modalities available today applied for in vivo studies of research animals, positron emission tomography (PET) is a technique that permits non-invasive use of positron labeled molecular imaging probes to image and assay biochemical processes of cellular function in the living subject. Imaging can be performed repeatedly before and after interventions and therefore allows the use of each animal as its own control. Many different positron labeled compounds have been and continue to be synthesized as probes that target a range of molecular targets within specific biochemical pathways. These molecular imaging probes are used in extremely low mass amounts, such that biological processes involving compounds in nanomolar concentration or lower can be imaged without disturbing the process. Biological processes from receptors and synthesis of transmitters in cell communication pathways, to metabolic processes and gene expression can be imaged. In the past, PET in animal research has been used extensively for studies of primates and larger animals. In recent years, the development of new detector technology has lowered the limits of spatial resolution. This has made it possible to use PET scanning for the study of the most important modern molecular biology model, the laboratory mouse. This paper presents some of the challenges facing small animal PET technology, provides an overview of the development of small animal PET systems, and discusses the current state of the art technology, some of its applications, as well as some future directions. PMID- 14538049 TI - Tantalum [18O]water target for the production of [18F]fluoride with high reactivity for the preparation of 2-deoxy-2-[18F]fluoro-D-glucose. AB - PURPOSE: To develop a new tantalum [18O]water target for the routine production of reactive, no-carrier-added [18F]fluoride ion in Curie amounts for the synthesis of radiopharmaceuticals. PROCEDURES: The tantalum target body was filled with 0.86 mL of 95% enriched [18O]water and irradiated with 10.2 MeV protons on target with beam currents of 26-40 microA for 60-90 min. [18F]Fluoride ion produced is trapped in an anion exchange resin cartridge and the expensive [18O]water is recovered for recycling. The [18F]fluoride ion is released from the resin by elution with a dilute solution of K(2)CO(3) and utilized in the synthesis of 2-deoxy-2-[18F]fluoro-D-glucose (FDG). RESULTS: Using the tantalum target, Curie levels of no-carrier-added [18F]fluoride ion (specific activity: >30,000 Ci/mmol) were reliably and reproducibly produced (n > 100), from which FDG was synthesized in 70% radiochemical yield. CONCLUSION: Tantalum [18O]water target body for the production of [18F]fluoride ion has the best characteristics from the standpoint of [18F]fluoride ion recovery and its radiochemical reactivity and low induced activation. This new target has the potential to replace the currently used silver and titanium [18O]targets for the routine production of [18F]fluoride ion. PMID- 14538050 TI - Indirect monitoring of endogenous gene expression by positron emission tomography (PET) imaging of reporter gene expression in transgenic mice. AB - PURPOSE: Repetitive imaging with microPET of endogenous albumin gene expression by using transgenic mice in which the Herpes Simplex Virus Type 1 thymidine kinase (HSV1-tk) reporter gene is driven by the albumin promoter (AL-HSV1-tk). METHODS: Transgenic mice were imaged repeatedly on a microPET scanner with approximately 200 microCi of 9-[4-[18F]fluoro-3-(hydroxymethyl)butyl]guanine (FHBG) (a substrate for HSV1-TK enzyme). Four transgenic mice were monitored for body weight, serum albumin, and imaged at the end of each of three dietary phases (17%, 0%, and 25% protein diet). Each phase last 14-21 days. The 0% protein diet has been reported previously to reduce albumin gene expression in rats. Twenty non-transgenic mice of the same strain followed a similar feeding schedule and were monitored for serum albumin, body weight, and sacrificed at various time points for determination of their GAPDH normalized albumin mRNA levels. RESULTS: Transgenic mice showed a relatively high FHBG signal from the liver region as expected. Variation of the mean FHBG signal in two mice with a fixed 17% protein diet over a four-month period was <19% s.d. The mean +/- s.e. FHBG liver standardized uptake value (SUV) in four transgenics went from 4.49 +/- 0.32 to 2.17 +/- 0.52 to 6.21 +/- 0.72 as the mice went through the three diets of 17%, 0%, and 25% sequentially. Non-transgenic mice showed GAPDH normalized albumin mRNA that went from 37.68 +/- 6.04 to 26.41 +/- 4.29 to 52.42 +/- 4.09. The FHBG SUV from transgenics was well correlated with GAPDH normalized albumin mRNA from non-transgenics (r(2) = 0.97) supporting that endogenous gene expression of albumin can be indirectly imaged with FHBG. CONCLUSION: Measuring correlated changes in albumin expression in wild type mice and HSV1-TK expression by microPET in transgenic mice in which the reporter gene is driven by the albumin promoter demonstrates that the HSV1-tk gene can be used to monitor, in living animals, modulated expression of transgenes. PMID- 14538051 TI - Kinetics of 3'-deoxy-3'-[F-18]fluorothymidine uptake and retention in dogs. AB - We have developed 3'-deoxy-3'-[F-18] fluorothymidine ([F-18]FLT) as an agent to image cellular proliferation with PET. Recent work has demonstrated that [F 18]FLT is stable to degradation and produces high contrast images of proliferating tissues and tumors. To increase our understanding for the use of this agent we have explored the kinetics of [F-18]FLT clearance from the blood and uptake into tissues in normal and tumor bearing dogs. The results indicate that [F-18]FLT is readily modeled in canines with a three-compartment model, with parameter k(3) representing phosphorylation by thymidine kinase. During the first 60 minutes, little loss was measured from the phosphorylated compartment, therefore parameter k(4) could not be differentiated from zero. The extraction of marrow from normal dogs was consistent with this model and demonstrated retention of phosphorylated [F-18]FLT. It is concluded that [F-18]FLT produces images of the DNA synthetic pathway by phosphorylation via thymidine kinase. This pathway can be readily modeled using a three-compartment model. PMID- 14538052 TI - FDG-PET in the pretherapeutic evaluation of primary squamous cell carcinoma of the oral cavity and the involvement of cervical lymph nodes. AB - PURPOSE: The diagnostic role of positron emission tomography (PET) with 2-deoxy-2 [18F]fluoro-D-glucose (FDG) in squamous cell carcinoma of the oral cavity is evaluated. PROCEDURES: In 38 patients, the results of FDG-PET, magnetic resonance imaging (MRI), computed tomography (CT), and ultrasound were compared. The standard uptake values (SUV) of FDG-PET were correlated to histopathological grading and DNA-image cytometry. RESULTS: In the case of lymph node metastases, the sensitivity of FDG-PET (93%) was higher than the sensitivity for the compared methods. The specificity was best for CT. SUVs of diploid tumor cell lines seemed to be lower than in non-diploid tumor cell lines. CONCLUSIONS: The high sensitivity and the high negative predictive value of PET may lead to more restrictive therapeutic regimens regarding lymph node metastases. Studies are necessary regarding possible relationships between glucose metabolism and tumor grading. PMID- 14538053 TI - Prognostic value of 2-deoxy-2-[F-18]fluoro-D-glucose positron emission tomography imaging for patients with prostate cancer. AB - PURPOSE: The purpose of this study was to investigate the prognostic value of measuring glucose metabolism of primary prostate cancer lesions, using 2-Deoxy-2 [F-18]Fluoro-D-Glucose positron emission tomography (FDG-PET). PROCEDURES: Forty two patients with prostate cancer were investigated with FDG-PET, and standardized uptake value (SUV) of the prostate was calculated. After PET study, radical prostatectomy was performed in 17 patients (RPT group), and endocrine therapy in 25 patients (ET group). Relapse-free survival curves were created by the Kaplan-Meier method. RESULTS: In the RPT group, the patients with high SUV had a poorer prognosis compared to those with low SUV (P = 0.033). In the ET group, the patients with high SUV were likely to have a poorer prognosis with low significance at a level of P = 0.087. CONCLUSIONS: FDG-PET appeared to have a defined prognostic value for patients with prostate cancer undergoing radical prostatectomy, and more patients need to be studied for patients undergoing endocrine therapy. PMID- 14538054 TI - 2-deoxy-2-[F-18]fluoro-D-glucose imaging with positron emission tomography for initial staging of Hodgkin's disease and lymphoma. AB - PURPOSE: To assess the accuracy of 2-Deoxy-2-[F-18] Fluoro-D-Glucose positron emission imaging (FDG-PET) for staging Hodgkin's disease (HD) and non-Hodgkin's lymphoma (NHL) compared to conventional staging (CS) and to evaluate the impact on patient management. METHODS: Forty-five consecutive patients with lymphoma underwent whole-body FDG-PET imaging for initial staging. Discordant lesions were verified with biopsy or clinical follow-up. The impact on staging and management was reviewed retrospectively. RESULTS: A total of 129 sites of disease were identified, and 88 of those were concordant. FDG-PET and conventional staging demonstrated 24 and 17 additional sites, respectively. FDG-PET correctly upstaged five patients and down-staged two patients (16% total), leading to a change in therapy in 6/45 (13%) patients. However, FDG-PET understaged three patients (7%), correctly staged by conventional staging modalities. Assuming that the addition of FDG-PET to conventional staging modalities is 100% accurate for staging lymphoma, the accuracy of FDG-PET alone was 91%, compared to 84% for conventional staging modalities. CONCLUSIONS: FDG-PET is a noninvasive and efficient imaging modality for staging patients with lymphoma and should be used in conjunction with conventional staging modalities, as they appear complementary. PMID- 14538055 TI - Monoclonal antibodies in organ transplantation. AB - Monoclonal antibody (mAb) technology has made possible the production of designer proteins, specifically reactive with almost any conceivable biological molecule. Using these reagents, the surface molecules on cells crucial for allograft rejection have been identified and described in detail. These structures can now be selectively targeted by mAb-based therapy in order to prevent rejection. For instance, the CD3 molecule, expressed on all mature T lymphocytes, triggers T cell activation, a key event in rejection. OKT3, an anti-CD3 mAb, disrupts T cell function and is now the agent of choice for the treatment of severe rejection episodes. MAbs targeting other T cell molecules are currently being investigated. Some of the most promising, the anti-CD4, anti-ICAM-1, and anti-interleukin 2 receptor mAbs, have already induced donor-specific tolerance in rodent models. These hosts accept permanently a genetically incompatible graft after only a limited period of mAb therapy. Interestingly, anti-ICAM-1 also diminishes the ischemic injury of preservation. The development of these new molecular agents, effectively directed to specific cellular targets, will likely play an increasingly important role in future clinical protocols, and perhaps finally provide a means to achieve long-term tolerance in human allograft recipients. PMID- 14538056 TI - Molecular mechanism of host specificity in legume-rhizobium symbiosis. AB - Rhizobium - legume symbiosis is a highly specific interaction between the two partners. Host specificity is evident at early stages of infection and results from multiple interactions involving signalling among bacteria and host plants. Host specific plant signals (flavanoids) convert the NodD protein to an active form and its binding with nod box initiates the transcription of inducible nod operons. Common nod genes (nodABC) code for an extracellular mitogenic Nod factor which is required for nodule organogenesis. Host specific genes (hsn) modify the Nod factor to induce root hair deformation on specific hosts. The structure of Nod factor controls host range distinction between species and biovars of rhizobia. Interactions of lectins and Exopolysaccharide/Lipopolysaccharide result in host specific attachment of Rhizobium and its subsequent invasion. Change in Expopolysaccharide structure by the transfer of hsn genes enables the Rhizobium to bind with heterologous host lectins. Conversely, changes in root lectins via gene manipulation enables the heterologous rhizobia to bind and initiate nodulation on heterologous hosts. Finally, host specific signals are required to initiate nitrogen fixation in nodules that are formed. PMID- 14538057 TI - Bioremoval of heavy metals by the use of microalgae. AB - Bioremoval, the use of biological systems for the removal of metal ions from polluted waters, has the potential to achieve greater performance at lower cost than conventional wastewater treatment technologies for metal removal. Bioremoval capabilities of microalgae have been extensively studied, and some commercial applications have been initiated. Although microalgae are not unique in their bioremoval capabilities, they offer advantages over other biological materials in some conceptual bioremoval process schemes. Selected microalgae strains, purposefully cultivated and processed for specific bioremoval applications, have the potential to provide significant improvements in dealing with the world-wide problems of metal pollution. In addition to strain selection, significant advances in the technology appear possible by improving biomass containment or immobilization techniques and by developing bioremoval process steps utilizing metabolically active microalgae cultures. The latter approach is especially attractive in applications where extremely low levels of residual metal ions are desired. This review summarizes the current literature, highlighting the potential benefits and problems associated with the development of novel algal based bioremoval processes for the abatement of heavy metal pollution. PMID- 14538058 TI - Applications of natural silk protein sericin in biomaterials. AB - Silk sericin is a natural macromolecular protein derived from silkworm Bombyx mori. During the various stages of producing raw silk and textile, sericin can be recovered for other uses. Also, sericin recovery reduces the environmental impact of silk manufacture. Sericin protein is useful because of its properties. The protein resists oxidation, is antibacterial, UV resistant, and absorbs and releases moisture easily. Sericin protein can be cross-linked, copolymerized, and blended with other macromolecular materials, especially artificial polymers, to produce materials with improved properties. The protein is also used as an improving reagent or a coating material for natural and artificial fibers, fabrics, and articles. The materials modified with sericin and sericin composites are useful as degradable biomaterials, biomedical materials, polymers for forming articles, functional membranes, fibers, and fabrics. PMID- 14538059 TI - Plant cell cultures: Chemical factories of secondary metabolites. AB - This review deals with the production of high-value secondary metabolites including pharmaceuticals and food additives through plant cell cultures, shoot cultures, root cultures and transgenic roots obtained through biotechnological means. Plant cell and transgenic hairy root cultures are promising potential alternative sources for the production of high-value secondary metabolites of industrial importance. Recent developments in transgenic research have opened up the possibility of the metabolic engineering of biosynthetic pathways to produce high-value secondary metabolites. The production of the pungent food additive capsaicin, the natural colour anthocyanin and the natural flavour vanillin is described in detail. PMID- 14538060 TI - Retroviral vectors for human gene delivery. AB - The potential for gene therapy to cure a wide range of diseases has lead to high expectations and a great increase in research efforts in this area. At present, viral vectors are the most efficient means of delivering a corrective gene into human cells. While a number of different viral vectors are under development, retroviral vectors are currently the most common type used in clinical trials today. However, the production of retroviral vectors for gene therapy applications faces a number of challenges. Of primary concern is the low titre of vector stocks produced by packaging cells in culture and the inherent instability of retroviral vector activity. The problems facing large-scale retroviral vector production are outlined in this review and the research efforts by a number of groups who have attempted to optimise production methods are presented. PMID- 14538061 TI - DNA markers and marker-assisted breeding for durable resistance to bacterial blight disease in rice. AB - Bacterial leaf blight caused by the bacterial pathogen Xanthomonas oryzae pv oryzae (Xoo) limits rice yield in all major rice-growing regions of the world, especially in irrigated lowland and rainfed conditions where predisposition factors favor disease development to epidemic proportions. Since bacterial pathogens are difficult to manage, development of host plant resistance is the most effective means of disease management. As many as 24 major genes conferring resistance to various races of the pathogen have been identified and utilized in rice breeding programs. However, large-scale and long-term cultivation of varieties carrying a single gene for resistance resulted in a significant shift in pathogen race frequency with consequent breakdown of resistance in these cultivars. To combat the problem of resistance breakdown, pyramiding of resistance genes into different cultivars is being carried out. Pyramiding of resistance genes is now possible with molecular markers that are developed for individual genes. This review discusses the various bacterial blight resistance genes identified and their corresponding molecular markers developed for breeding durable resistance into modern rice cultivars. PMID- 14538064 TI - Two-phase partitioning bioreactors in fermentation technology. AB - The two-phase partitioning bioreactor concept appears to have a great potential in enhancing the productivity of many bioprocesses. The proper selection of an organic solvent is the key to successful application of this approach in industrial practice. The integration of fermentation and a primary product separation step has a positive impact on the productivity of many fermentation processes. The controlled substrate delivery from the organic to the aqueous phase opens a new area of application of this strategy to biodegradation of xenobiotics. In this review, the most recent advances in the application of two liquid phase partitioning bioreactors for product or substrate partitioning are discussed. Modeling and performance optimization studies related to those bioreactor systems are also reviewed. PMID- 14538065 TI - Management strategies for agricultural biotechnology in small countries. A case study of Israel. AB - Agricultural biotechnology is concentrated in four major countries. This paper suggests strategies for developing it in small countries, based on analysis of the world trends and the characteristics of small countries. Israel is presented as a specific case study. The main relevant trends are domination by big companies, consumer concerns on genetically modified foods, and focusing on consumer benefits and specific market niches. Small countries' disadvantages include companies that are too small to benefit fully from research, difficulty in raising funds, lack of infrastructures and experienced management personnel, and public sector research organizations that are unsuitable for commercializing research. The recommended strategies include: developing a large number of low volume products and small market niches, forming partnerships with intermediaries (such as food companies), specializing in intermediate products (such as the seed or the gene patent), and conducting market research and cost-benefit analysis in advance. Additional strategies include developing benefits that are unique to genetically modified foods and focusing on benefits specifically for consumers who accept genetically modified foods, rather than on benefits for the average consumer. A national representative organization could buy and rent out expensive equipment, finance specific projects in return for the commercial rights, and perform collective marketing research and marketing. Israel has the advantages of a successful agricultural sector and complementary scientific research, and should focus on those fruits, vegetables, and flowers for which it already has the experience and infrastructure. PMID- 14538068 TI - Advances in pretargeting biotechnology. AB - A major focus of current drug research is to improve drug targeting to internal target sites such as to solid tumors or specific organs. The objective of drug targeting, especially for cancer chemotherapy and radioimmunotherapy, is to enhance the effectiveness of the drug by concentrating it at the target site and minimizing its effects in nontarget sites. Although tumor targeting has been obtained with large long-circulating radiolabeled antibody molecules, normal organ activity, especially in the blood kidneys, liver, and bone marrow is a significant problem. Over the last 20 years, studies to improve the therapeutic use of antibodies have included the use of antibody fragments, chase molecules, metabolizable linkers, antibody-directed enzyme prodrugs (ADEPT), local delivery, and pretargeting. Here, we will review the most interesting recent advances in pretargeting biotechnology. PMID- 14538069 TI - Scanning force microscopy in the applied biological sciences. AB - Fifteen years after its invention, the scanning force microscope (SFM) is rooted deep in the biological sciences. Here we discuss the use of SFM in biotechnology and biomedical research. The spectrum of applications reviewed includes imaging, force spectroscopy and mapping, as well as sensor applications. It is our hope that this review will be useful for researchers considering the use of SFM in their studies but are uncertain about its scope of capabilities. For the benefit of readers unfamiliar with SFM technology, the fundamentals of SFM imaging and force measurement are also briefly introduced. PMID- 14538072 TI - Debittering of protein hydrolyzates. AB - Enzymatic hydrolysis of proteins frequently results in bitter taste, which is due to the formation of low molecular weight peptides composed of mainly hydrophobic amino acids. Methods for debittering of protein hydrolyzates include selective separation such as treatment with activated carbon, extraction with alcohol, isoelectric precipitation, chromatography on silica gel, hydrophobic interaction chromatography, and masking of bitter taste. Bio-based methods include further hydrolysis of bitter peptides with enzymes such as aminopeptidase, alkaline/neutral protease and carboxypeptidase, condensation reactions of bitter peptides using protease, and use of Lactobacillus as a debittering starter adjunct. The causes for the production of bitter peptides in various food protein hydrolyzates and the development of methods for the prevention, reduction, and elimination of bitterness as well as masking of bitter taste in enzymatic protein hydrolyzates are presented. PMID- 14538073 TI - Extracellular biopolymeric flocculants. Recent trends and biotechnological importance. AB - Many microorganisms secrete extracellular biopolymeric flocculants (EBFs) in the culture broth. This work reviews the development of EBF research and applications. Aspects discussed include a comparison of the chemical and biological flocculating agents, isolation of EBF-producing microorganisms, culture conditions, mechanisms of flocculation, the chemical structure of EBFs, and the role of physicochemical factors in the flocculating activity. PMID- 14538074 TI - Biotechnological production of prostaglandin. AB - Prostaglandins (PGs) are the oxidation products of PG endoperoxide (PGH) synthase and other tissue enzymes. They occur in a tissue-specific manner and act as local hormones. Biotechnological production of PGs has been of interest, but not yet fully established. Biological tissues have been used as PG sources, but this disturbs ecological balance, and the cost of production is very high for commercial purposes. On the other hand, various microorganisms have been shown to synthesize them de novo, or biotransform precursors to active molecules, but these processes have not been further evaluated. Using mammalian enzymes in free or immobilized form is a promising new approach to synthesize PG from fatty acid substrates. Rapid enzyme inactivation during the catalysis is the main problem to be solved. Optimization of factors in the reactions and the design of special reactors that will allow removal of products continuously from the reaction medium without affecting enzyme activity need immediate attention from researchers and the pharmaceutical industry. PMID- 14538076 TI - Biotechnological uses of archaeal extremozymes. AB - Archaea have developed a variety of molecular strategies to survive the often harsh environments in which they exist. Although the rules that allow archaeal enzymes to fulfill their catalytic functions under extremes of salinity, temperature or pressure are not completely understood, the stability of these extremophilic enzymes, or extremozymes, in the face of adverse conditions has led to their use in a variety of biotechnological applications in which such tolerances are advantageous. In the following, examples of commercially important archaeal extremozymes are presented, potentially useful archaeal extremozyme sources are identified and solutions to obstacles currently hindering wider use of archaeal extremozymes are discussed. PMID- 14538077 TI - Gene therapy. Therapeutic approaches and implications. AB - The present article is an overview of gene therapy with an emphasis on different approaches and its implications in the clinic. Genetic interventions have been applied to the diagnosis of and therapy for an array of human diseases. The initial concept of gene therapy was focused on the treatment of genetic diseases. Subsequently, the field of gene therapy has been expanded, with a major focus on cancer. Although the results of early gene therapy-based clinical trials have been encouraging, there is a need for gene delivery vectors that feature reduced immunogenicity and improved targeting ability. The results of phases I/II clinical trials have suggested the important role of gene therapy as a versatile and powerful treatment tool, especially for human cancers. One reasonable expectation is that performing gene therapy at an earlier stage in the disease process or for minimal residual disease may be more advantageous. PMID- 14538080 TI - Manipulating DNA sequence data. PMID- 14538078 TI - The prospects of modifying the antimicrobial properties of milk. AB - Milk contains a variety of substances, which inhibit the infection of pathogens. This is of benefit to the mother, safeguarding the integrity of the lactating mammary gland, but also of huge importance for protection of the suckling offspring. The antimicrobial substances in milk can be classified into two categories. First, nonspecific, broad-spectrum defense substances, which have evolved over long periods of time, and secondly, substances like antibodies, which are specifically directed against particular pathogens and have developed during the mother's lifetime. Substances in both categories may be targets for biological intervention and manipulation with the goal of improving the antimicrobial properties of milk. These alterations of milk composition have applications in human as well as in animal health. PMID- 14538081 TI - Is only the cow mad? PMID- 14538082 TI - Biotransformations using plant cells, organ cultures and enzyme systems: current trends and future prospects. AB - Plants are valuable sources of a variety of chemicals including drugs, flavours, pigments and agrochemicals. Some of the biochemical reactions occurring in plant cells are complex and cannot be achieved by synthetic routes. In vitro plant cell and organ cultures and plant enzymes act as suitable biocatalysts to perform these complex reactions. A wide variety of chemical compounds including aromatics, steroids, alkaloids, coumarins and terpenoids can undergo biotransformations using plant cells, organ cultures and enzymes. The biocatalyst mediated reactions are regiospecific and stereospecific. Reaction types include oxidations, reductions, hydroxylations, methylations, acetylations, isomerizations, glycosylations and esterfications. Genetic manipulation approaches to biotransformation offer great potential to express heterologous genes and to clone and overexpress genes for key enzymes. Biotransformation efficiencies can further be improved using molecular techniques involving site directed mutagenesis and gene manipulation for substrate specificity. PMID- 14538083 TI - Glycerol production by microbial fermentation: a review. AB - Microbial production of glycerol has been known for 150 years, and glycerol was produced commercially during World War I. Glycerol production by microbial synthesis subsequently declined since it was unable to compete with chemical synthesis from petrochemical feedstocks due to the low glycerol yields and the difficulty with extraction and purification of glycerol from broth. As the cost of propylene has increased and its availability has decreased especially in developing countries and as glycerol has become an attractive feedstock for production of various chemicals, glycerol production by fermentation has become more attractive as an alternative route. Substantial overproduction of glycerol by yeast from monosaccharides can be obtained by: (1) forming a complex between acetaldehyde and bisulfite ions thereby retarding ethanol production and restoring the redox balance through glycerol synthesis; (2) growing yeast cultures at pH values near 7 or above; or (3) using osmotolerant yeasts. In recent years, significant improvements have been made in the glycerol production using osmotolerant yeasts on a commercial scale in China. The most outstanding achievements include: (1) isolation of novel osmotolerant yeast strains producing up to 130 g/L glycerol with yields up to 63% and the productivities up to 32 g/(L day); (2) glycerol yields, productivities and concentrations in broth up to 58%, 30 g/(L day) and 110-120 g/L, respectively, in an optimized aerobic fermentation process have been attained on a commercial scale; and (3) a carrier distillation technique with a glycerol distillation efficiency greater than 90% has been developed. As glycerol metabolism has become better understood in yeasts, opportunities will arise to construct novel glycerol overproducing microorganisms by metabolic engineering. PMID- 14538085 TI - Strategy for minimization of excess sludge production from the activated sludge process. AB - Increased attention has been given to minimization of sludge production from activated sludge process since environmental regulations are being more and more stringent in relation to excess sludge disposal. In a biological process, the more organic carbon utilized in carbon dioxide production, the fewer sludge produced, and vice versa. This paper, therefore, reviews strategies developed for minimization of excess sludge production, such as oxic-settling-anaerobic process, high dissolved oxygen process, uncoupler-containing activated sludge process, ozonation-combined activated sludge process, control of sludge retention time and biodegradation of sludge in membrane-assisted reactor. In these modified activated sludge processes, excess sludge production can be reduced by 20-100% without significant effect on process efficiency and stability. It is expected that this paper would be helpful for researchers and engineers to develop novel and efficient operation strategy to minimize sludge production from biological systems. PMID- 14538086 TI - Polymerized lipid vesicles as colorimetric biosensors for biotechnological applications. AB - Supramolecular chemical assemblies composed of polydiacetylene (PDA) exhibit rapid colorimetric transitions upon specific interactions with a variety of biological analytes in aqueous solutions. Among the analytes that give rise to the unique blue-red color changes are lipophilic enzymes, antibacterial peptides, ions, antibodies, and membrane penetration enhancers. The chemical assemblies include conjugated PDA, responsible for the chromatic transitions, and the molecular recognition elements, which are either chemically or physically associated with the PDA. Thus, by incorporation of specific recognition elements, the system can be designed in ways allowing for highly selective identification of analytes. In particular, receptors and epitopes can be incorporated within the sensor assembly, which then determine the specificity of the colorimetric transitions. The PDA-based molecular assemblies are robust and can be readily applied to diagnosis of physiological molecules and for rapid screening of chemical and biological libraries, for example, in 96 well-plate platforms. PMID- 14538087 TI - Microbial leaching of metals from sulfide minerals. AB - Microorganisms are important in metal recovery from ores, particularly sulfide ores. Copper, zinc, gold, etc. can be recovered from sulfide ores by microbial leaching. Mineral solubilization is achieved both by 'direct (contact) leaching' by bacteria and by 'indirect leaching' by ferric iron (Fe(3+)) that is regenerated from ferrous iron (Fe(2+)) by bacterial oxidation. Thiobacillus ferrooxidans is the most studied organism in microbial leaching, but other iron- or sulfide/sulfur-oxidizing bacteria as well as archaea are potential microbial agents for metal leaching at high temperature or low pH environment. Oxidation of iron or sulfur can be selectively controlled leading to solubilization of desired metals leaving undesired metals (e.g., Fe) behind. Microbial contribution is obvious even in electrochemistry of galvanic interactions between minerals. PMID- 14538089 TI - Bacterial promotion of plant growth. PMID- 14538090 TI - Biotechnological applications of phage and cell display. AB - In recent years, the use of surface-display vectors for displaying polypeptides on the surface of bacteriophage and bacteria, combined with in vitro selection technologies, has transformed the way in which we generate and manipulate ligands, such as enzymes, antibodies and peptides. Phage display is based on expressing recombinant proteins or peptides fused to a phage coat protein. Bacterial display is based on expressing recombinant proteins fused to sorting signals that direct their incorporation on the cell surface. In both systems, the genetic information encoding for the displayed molecule is physically linked to its product via the displaying particle. Using these two complementary technologies, we are now able to design repertoires of ligands from scratch and use the power of affinity selection to select those ligands having the desired (biological) properties from a large excess of irrelevant ones. With phage display, tailor-made proteins (fused peptides, antibodies, enzymes, DNA-binding proteins) may be synthesized and selected to acquire the desired catalytic properties or affinity of binding and specificity for in vitro and in vivo diagnosis, for immunotherapy of human disease or for biocatalysis. Bacterial surface display has found a range of applications in the expression of various antigenic determinants, heterologous enzymes, single-chain antibodies, and combinatorial peptide libraries. This review explains the basis of phage and bacterial surface display and discusses the contributions made by these two leading technologies to biotechnological applications. This review focuses mainly on three areas where phage and cell display have had the greatest impact, namely, antibody engineering, enzyme technology and vaccine development. PMID- 14538091 TI - Developments in odour control and waste gas treatment biotechnology: a review. AB - Waste and wastewater treatment processes produce odours, which can cause a nuisance to adjacent populations and contribute significantly to atmospheric pollution. Sulphurous compounds are responsible for acid rain and mist; many organic compounds of industrial origin contribute to airborne public health concerns, as well as environmental problems. Waste gases from industry have traditionally been treated using physicochemical processes, such as scrubbing, adsorption, condensation, and oxidation, however, biological treatment of waste gases has gained support as an effective and economical option in the past few decades. One emergent technique for biological waste gas treatment is the use of existing activated sludge plants as bioscrubbers, thus treating the foul air generated by other process units of the wastewater treatment system on site, with no requirement for additional units or for interruption of wastewater treatment. Limited data are available regarding the performance of activated sludge diffusion of odorous air in spite of numerous positive reports from full-scale applications in North America. This review argues that the information available is insufficient for precise process design and optimization, and simultaneous activated sludge treatment of wastewater and airborne odours could be adopted worldwide. PMID- 14538092 TI - A critical examination of current HIV therapies. AB - This review critically examines the current methods of eliminating and preventing human immunodeficiency virus (HIV) infection. It illustrates both the experimental and practical limitations that each approach faces, and how they may be overcome. An overview of the HIV, including its structure and life cycle is presented. Subsequently, the two main methods of post-infection treatment, drug and gene therapy are outlined. The development of HIV vaccination is discussed with an analysis of conventional vaccination techniques leading into the novel approaches. The final option examined describes the potential for a combined vaccination regimen. Finally, the question of why these approaches have met with little success is addressed. This includes practical research limitations, as well as an examination of the qualities of HIV that make it so elusive. PMID- 14538093 TI - Production of transgenic rice with agronomically useful genes: an assessment. AB - Rice is the most important food crop in tropical and subtropical regions of the world. Yield enhancement to increase rice production is one of the essential strategies to meet the demand for food of the growing population. Both abiotic and biotic features limit adversely the productivity of rice growing areas. Conventional breeding has been an effective means for developing high yielding varieties, however; it is associated with its own limitations. It is envisaged that recent trends in biotechnology can contribute to the agronomic improvement of rice in terms of yield and nutritional quality as a supplement to traditional breeding methods. Genetic transformation of rice has demonstrated numerous important opportunities resulting in the genetic improvement of existing elite rice varieties and production of new plant types. Significant advances have been made in the genetic engineering of rice since the first transgenic rice plant production in the late 1980s. Several gene transfer protocols have been employed successfully for the introduction of foreign genes to rice. In more than 60 rice cultivars belonging to indica, japonica, javanica, and elite African cultivars, the protocol has been standardized for transgenic rice production. Selection and use of appropriate promoters, selectable markers, and reporter genes has been helpful for development of efficient protocols for transgenic rice in a number of rice cultivars. The present review is an attempt to assess the current state of development in transgenic rice for the transfer of agronomically useful genes, emphasizing the application and future prospects of transgenic rice production for the genetic improvement of this food crop. PMID- 14538094 TI - Marine biotechnology - a neglected resource. PMID- 14538095 TI - Xanthan gum: production, recovery, and properties. AB - Xanthan gum is a microbial polysaccharide of great commercial significance. This review focuses on various aspects of xanthan production, including the producing organism Xanthomonas campestris, the kinetics of growth and production, the downstream recovery of the polysaccharide, and the solution properties of xanthan. PMID- 14538096 TI - Integrated processing of biotechnology products. AB - Integrated bioprocessing in which a potentially inhibitory product is continuously removed from the fermentation broth as it is produced, has important advantages in improving yield and conversion relative to conventional processes. This review discusses integrated processing for ethanol, butanol, organic acids, antibiotics, and other products. A variety of recovery operations can be used to isolate the product, as discussed. Use of some of the available options is compared. PMID- 14538097 TI - Value-added food: single cell protein. AB - The alarming rate of population growth has increased the demand for food production in third-world countries leading to a yawning gap in demand and supply. This has led to an increase in the number of hungry and chronically malnourished people. This situation has created a demand for the formulation of innovative and alternative proteinaceous food sources. Single cell protein (SCP) production is a major step in this direction. SCP is the protein extracted from cultivated microbial biomass. It can be used for protein supplementation of a staple diet by replacing costly conventional sources like soymeal and fishmeal to alleviate the problem of protein scarcity. Moreover, bioconversion of agricultural and industrial wastes to protein-rich food and fodder stocks has an additional benefit of making the final product cheaper. This would also offset the negative cost value of wastes used as substrate to yield SCP. Further, it would make food production less dependent upon land and relieve the pressure on agriculture. This article reviews diversified aspects of SCP as an alternative protein-supplementing source. Various potential strains and substrates that could be utilized for SCP production are described. Nutritive value and removal of nucleic acids and toxins from SCP as a protein-supplementing source are discussed. New processes need to be exploited to improve yield. In that direction the solid state fermentation (SSF) method and its advantages for SCP production are highlighted. PMID- 14538098 TI - Plants as 'chemical factories' for the production of polyunsaturated fatty acids. AB - Polyunsaturated fatty acids (PUFAs) are valuable products because of their involvement in several aspects of human health. Market demand for most PUFAs is growing continually and current sources are considered insufficient for satisfying this demand; alternative sources are actively sought after. Oilseed plants can be a potential source of PUFAs if they are appropriately gene engineered. Most of the basic tools for genetic engineering of oilseed plants for giving them the ability to produce PUFAs are already developed. Here we review the prospects of genetic engineering of oilseed plants for producing some valuable long-chain polyunsaturated fatty acids. Genetic transformation for GLA production seems to be a near-term possibility, but gene engineering seems considerably more difficult for the other long-chain PUFAs. Nevertheless, with the current rapid pace of biotechnological advancement, the remaining difficulties may be surmounted in the near future. PMID- 14538099 TI - Small bugs, big business: the economic power of the microbe. AB - The versatility of microbial biosynthesis is enormous. The most industrially important primary metabolites are the amino acids, nucleotides, vitamins, solvents, and organic acids. Millions of tons of amino acids are produced each year with a total multibillion dollar market. Many synthetic vitamin production processes are being replaced by microbial fermentations. In addition to the multiple reaction sequences of fermentations, microorganisms are extremely useful in carrying out biotransformation processes. These are becoming essential to the fine chemical industry in the production of single-isomer intermediates. Microbially produced secondary metabolites are extremely important to our health and nutrition. As a group, they have tremendous economic importance. The antibiotic market amounts to almost 30 billion dollars and includes about 160 antibiotics and derivatives such as the beta-lactam peptide antibiotics, the macrolide polyketide erythromycin, tetracyclines, aminoglycosides and others. Other important pharmaceutical products produced by microrganisms are hypocholesterolemic agents, enzyme inhibitors, immunosuppressants and antitumor compounds, some having markets of over 1 billion dollars per year. Agriculturally important secondary metabolites include coccidiostats, animal growth promotants, antihelmintics and biopesticides. The modern biotechnology industry has made a major impact in the business world, biopharmaceuticals (recombinant protein drugs, vaccines and monoclonal antibodies) having a market of 15 billion dollars. Recombinant DNA technology has also produced a revolution in agriculture and has markedly increased markets for microbial enzymes. Molecular manipulations have been added to mutational techniques as means of increasing titers and yields of microbial procresses and in discovery of new drugs. Today, microbiology is a major participant in global industry. The best is yet to come as microbes move into the environmental and energy sectors. PMID- 14538100 TI - Cellulases and related enzymes in biotechnology. AB - Basic and applied research on microbial cellulases, hemicellulases and pectinases has not only generated significant scientific knowledge but has also revealed their enormous potential in biotechnology. At present, cellulases and related enzymes are used in food, brewery and wine, animal feed, textile and laundry, pulp and paper industries, as well as in agriculture and for research purposes. Indeed, the demand for these enzymes is growing more rapidly than ever before, and this demand has become the driving force for research on cellulases and related enzymes. The present article is an overview of the biotechnological state of-the-art for cellulases and related enzymes. PMID- 14538101 TI - Monoclonal antibodies and therapy of human cancers. AB - This survey is an overview of the applications of murine, humanized and recombinant monoclonal antibodies for in vivo diagnostic and therapeutic applications. Monoclonal antibodies (mAb) have been applied to the diagnosis and therapy of an array of human diseases. The initial failures of early clinical trials have been overcome through the production of a new generation of mAb which features reduced immunogenicity and improved targeting abilities. The early models of mAb therapy were focused on enhancing the cytolytic mechanisms against the tumor cells. More recently, successful mAb-based therapies were targeted to molecules involved in the regulation of growth of cancer cells. This has highlighted the relevance of understanding receptor-mediated signaling events, and may provide new opportunities for anti-tumor antibody targeting. Despite all the difficulties, clinical data is outlining an increasingly significant role for antibody-mediated cancer therapy as a versatile and powerful instrument in cancer treatment. One reasonable expectation is that treatment at an earlier stage in the disease process or in minimal residual disease may be more advantageous. PMID- 14538102 TI - Alpha-L-arabinofuranosidases: biochemistry, molecular biology and application in biotechnology. AB - Interest in the alpha-L-arabinofuranosidases has increased in recent years because of their application in the conversion of various hemicellulosic substrates to fermentable sugars for subsequent production of fuel alcohol. Xylanases, in conjunction with alpha-L-arabinofuranosidases and other accessory enzymes, act synergistically to degrade xylan to component sugars. The induction of alpha-L-arabinofuranosidase production, physico-chemical characteristics, substrate specificity, and molecular biology of the enzyme are described. The current state of research and development of the arabinofuranosidases and their role in biotechnology are presented. PMID- 14538104 TI - Are microbes intelligent beings?: An assessment of cybernetic modeling. AB - Microorganisms growing in a multi-substrate medium have different and varying preferences for the various components of the medium. The preferences depend on the operating conditions and the substrates may be utilized sequentially or simultaneously. Sometimes an organism may change its preferences among substrates and/or switch between sequential and simultaneous utilization. These aspects are difficult to describe through models based on chemical and physical laws alone. Cybernetic modeling ascribes to microorganisms the ability to perceive their environment (i.e. the growth medium) and make 'intelligent' choices regarding substrate utilization to maximize an objective, which is usually the growth rate. This article reviews the development of cybernetic modeling since it began in 1982. Different workers have suggested different perspectives of how microbes make optimal use of their resources. These are discussed and future directions for improvement are indicated. PMID- 14538105 TI - Advances in enzymatic transformation of penicillins to 6-aminopenicillanic acid (6-APA). AB - This article elaborates on the important recent developments in the enzymatic transformation of penicillins to 6-aminopenicillanic acid (6-APA), which is the basic raw material for the industrial production of semisynthetic penicillins such as amoxycillin and ampicillin. Particular emphasis is placed on the improvements in purification, stability, and immobilization of the enzymes, (i.e. penicillin acylases) used for these transformations. PMID- 14538106 TI - Microencapsulation of microbial cells. AB - The high level of biocatalysts such as microbial cells and enzymes plays an important role in increasing the productivity of a bioreactor. The beads entrapped with microbial cells are not strong enough for long-term use. The small void space of polymer matrix and the leakage of cells limit a final cell loading in the beads. The recent success of encapsulating microbial cells makes it possible to prepare dense biocatalyst composed of recombinant microbial cells. In addition to encapsulating microbial cells, immobilization of animal and plant cells in capsules is also briefly described. PMID- 14538107 TI - The impact of genetic modification of human foods in the 21st century: a review. AB - Genetic engineering of food is the science which involves deliberate modification of the genetic material of plants or animals. It is an old agricultural practice carried on by farmers since early historical times, but recently it has been improved by technology. Many foods consumed today are either genetically modified (GM) whole foods, or contain ingredients derived from gene modification technology. Billions of dollars in U.S. food exports are realized from sales of GM seeds and crops. Despite the potential benefits of genetic engineering of foods, the technology is surrounded by controversy. Critics of GM technology include consumer and health groups, grain importers from European Union (EU) countries, organic farmers, environmentalists, concerned scientists, ethicists, religious rights groups, food advocacy groups, some politicians and trade protectionists. Some of the specific fears expressed by opponents of GM technology include alteration in nutritional quality of foods, potential toxicity, possible antibiotic resistance from GM crops, potential allergenicity and carcinogenicity from consuming GM foods. In addition, some more general concerns include environmental pollution, unintentional gene transfer to wild plants, possible creation of new viruses and toxins, limited access to seeds due to patenting of GM food plants, threat to crop genetic diversity, religious, cultural and ethical concerns, as well as fear of the unknown. Supporters of GM technology include private industries, research scientists, some consumers, U.S. farmers and regulatory agencies. Benefits presented by proponents of GM technology include improvement in fruit and vegetable shelf-life and organoleptic quality, improved nutritional quality and health benefits in foods, improved protein and carbohydrate content of foods, improved fat quality, improved quality and quantity of meat, milk and livestock. Other potential benefits are: the use of GM livestock to grow organs for transplant into humans, increased crop yield, improvement in agriculture through breeding insect, pest, disease, and weather resistant crops and herbicide tolerant crops, use of GM plants as bio-factories to yield raw materials for industrial uses, use of GM organisms in drug manufacture, in recycling and/or removal of toxic industrial wastes. The potential risks and benefits of the new technology to man and the environment are reviewed. Ways of minimizing potential risks and maximizing the benefits of GM foods are suggested. Because the benefits of GM foods apparently far outweigh the risks, regulatory agencies and industries involved in GM food business should increase public awareness in this technology to enhance worldwide acceptability of GM foods. This can be achieved through openness, education, and research. PMID- 14538108 TI - Production, purification, and characterization of the debittering enzyme naringinase. AB - This review discusses the debittering enzyme naringinase and its essential role in the commercial processing of citrus fruit juice. Applications of this enzyme in other areas are identified. Characterization of the enzyme is detailed and its immobilized preparations are discussed. Production of microbial naringinase by fermentation is described. PMID- 14538109 TI - Production of nitrogen oxide and dinitrogen oxide by autotrophic nitrifiers. AB - Autotrophic nitrifiers have been shown to produce nitrogen oxide and dinitrogen oxide under oxic conditions. Dinitrogen oxide is produced mainly during nitrite reduction (i.e. aerobic denitrification) whereas nitrogen oxide is produced during both aerobic denitrification and as a result of chemodenitrification. Oxygen is the single most influential environmental factor affecting the production of nitrogen and dinitrogen oxides; a decrease in oxygen can result in a several-fold increase in nitrogen oxide and dinitrogen oxide production. Emission of nitrogen oxide and dinitrogen oxide from wastewater treatment plants and fertilized soils is well documented; however, only recently have the contributions from such environments to the global nitrogen and dinitrogen oxide budget been considered. PMID- 14538111 TI - The production of antibodies in plants: an idea whose time has come? PMID- 14538112 TI - In vitro manipulation and propagation of medicinal plants. AB - Well developed techniques are currently available to help growers meet the demand of the pharmaceutical industry in the next century. These protocols are designed to provide optimal levels of carbohydrates, organic compounds (vitamins), mineral nutrients, environmental factors (e.g. light, gaseous environment, temperature, and humidity) and growth regulators required to obtain high regeneration rates of many plant species in vitro and thereby facilitate commercially viable micropropagation. Well-defined cell culture methods have also been developed for the production of several important secondary products. An overview of the regeneration of medicinal plants by direct and indirect organogenesis and by somatic embryogenesis from various types of explants is presented, and the use of these techniques combined with other biotechnological approaches to improve medicinal plants through somaclonal variation and genetic transformation is reviewed. PMID- 14538113 TI - Genetic immobilization of proteins on the yeast cell surface. AB - A genetic system has been exploited to immobilize proteins in their active and functional forms on the cell surface of yeast, Saccharomyces cerevisiae. DNAs encoding proteins with a secretion signal peptide were fused with the genes encoding yeast agglutinins, a- and alpha-type proteins involved in mating. The fusion gene was introduced into S. cerevisiae and expressed under the control of several promoters. Appearance of the fused proteins expressed on the cell surface was demonstrated biochemically and by immunofluorescence and immunoelectron microscopy techniques. Alpha-galactosidase from Cyamopsis tetragonoloba seeds, peptide libraries including scFv and variable regions of the T cell receptor from mammalian cells have been successfully immobilized on the yeast cell wall in the active form. Recently, surface-engineered yeasts have been constructed by immobilizing the enzymes and a functional protein, for example, green fluorescent protein (GFP) from Aequorea victoria. The yeasts were termed 'arming yeasts' with biocatalysts or functional proteins. Such arming cells displaying glucoamylase from Rhizopus oryzae and alpha-amylase from Bacillus stearothermophilus, or carboxymethylcellulase and beta-glucosidase from Aspergillus acleatus, could assimilate starch or cellooligosaccharides as the sole carbon source, although S. cerevisiae cannot intrinsically assimilate these substrates. GFP-arming cells can emit green fluorescence from the cell surface in response to the environmental conditions. The approach described in this review will enable us to endow living cells, including yeast cells, with novel additional abilities and to open new dimensions in the field of biotechnology. PMID- 14538115 TI - Recent developments in the biotechnology of Bacillus thuringiensis. PMID- 14538116 TI - Transgenic hairy roots. recent trends and applications. AB - Agrobacterium rhizogenes causes hairy root disease in plants. The neoplastic roots produced by A. rhizogenes infection is characterized by high growth rate and genetic stability. These genetically transformed root cultures can produce higher levels of secondary metabolites or amounts comparable to that of intact plants. Hairy root cultures offer promise for production of valuable secondary metabolites in many plants. The main constraint for commercial exploitation of hairy root cultures is their scaling up, as there is a need for developing a specially designed bioreactor that permits the growth of interconnected tissues unevenly distributed throughout the vessel. Rheological characteristics of heterogeneous system should also be taken into consideration during mass scale culturing of hairy roots. Development of bioreactor models for hairy root cultures is still a recent phenomenon. It is also necessary to develop computer aided models for different parameters such as oxygen consumption and excretion of product to the medium. Further, transformed roots are able to regenerate genetically stable plants as transgenics or clones. This property of rapid growth and high plantlet regeneration frequency allows clonal propagation of elite plants. In addition, the altered phenotype of hairy root regenerants (hairy root syndrome) is useful in plant breeding programs with plants of ornamental interest. In vitro transformation and regeneration from hairy roots facilitates application of biotechnology to tree species. The ability to manipulate trees at a cellular and molecular level shows great potential for clonal propagation and genetic improvement. Transgenic root system offers tremendous potential for introducing additional genes along with the Ri T-DNA genes for alteration of metabolic pathways and production of useful metabolites or compounds of interest. This article discusses various applications and perspectives of hairy root cultures and the recent progress achieved with respect to transformation of plants using A. rhizogenes. PMID- 14538117 TI - Exploitation of plants for the removal of organics in environmental remediation. AB - This review concentrates on the description of various phytoremediation technologies, paying special attention to removal of organics and the application of in vitro systems for basic research in the role of plants for the remediation of contaminated sites or flows, and in the improvement of their effectiveness. Various aspects of xenobiotic metabolism in plant cells, the role of enzymes involved, and the cooperation with rhizospheric microorganisms accelerating remediation of organics are shown. Application of this approach as well as the possibility of introduction of foreign genes into plant genome that can enhance the rate of the bioremediation are discussed. PMID- 14538118 TI - cDNA microarray technology and its applications. AB - The cDNA microarray is the most powerful tool for studying gene expression in many different organisms. It has been successfully applied to the simultaneous expression of many thousands of genes and to large-scale gene discovery, as well as polymorphism screening and mapping of genomic DNA clones. It is a high throughput, highly parallel RNA expression assay technique that permits quantitative analysis of RNAs transcribed from both known and unknown genes. This technique provides diagnostic fingerprints by comparing gene expression patterns in normal and pathological cells, and because it can simultaneously track expression levels of many genes, it provides a source of operational context for inference and predication about complex cell control systems. This review describes this recently developed cDNA microarray technology and its application to gene discovery and expression, and to diagnostics for certain diseases. PMID- 14538121 TI - Biotechnology products and European consumers. AB - More than 100 interviews conducted during 1997 with European food manufacturers and retailers, trade associations, government departments, consumer groups, environmental organizations and some individual academic scientists revealed how differences in the perceived attitudes of consumers gave rise to varying approaches by suppliers to the possible introduction of transgenic foods. European consumers generally are not against the pharmaceutical products of biotechnology but are much less willing to accept food and food ingredients, especially when derived from genetically modified plants. Objections are mainly based on fears for the health and safety of the consumer, worries about the possibility of deleterious effects on the environment, and a range of moral and ethical concerns often deriving from a distaste, however expressed, at the concept of interfering with nature. Consumer understanding of the science underlying biotechnology is patchy; in no country does more than a small proportion of the population claim a good grasp. Partly no doubt as a consequence of these attitudes, the introduction of genetically modified foods into Europe has occurred slowly and, during the period of this study, perhaps only in the Netherlands and the UK. PMID- 14538122 TI - Novel biotechnological approaches in environmental remediation research. AB - Two novel approaches, the use of Agrobacterium-transformed plant roots and mycelia cultures of fungi, are considered as research tools in the study of the remediation of soil, groundwater, and biowastes. Transformed roots are excellent model systems for screening higher plants that are tolerant of various inorganic and organic pollutants, and for determining the role of the root matrix in the uptake and further metabolism of contaminants. Edible and/or medicinal fungi may also be natural environmental remediators. Liquid cultures of fungal mycelia are appropriate model systems with which to commence screening and biochemical studies in this under-researched area of biotransformation. PMID- 14538125 TI - Applications of neural networks to recovery of biological products. AB - Artificial neural networks (ANN) are being applied to recovery of products from fermentation broths. Recovery methods for which mathematical models are complex or non-existent are particularly suitable for control and analysis by ANNs. Use and potential of artificial neural networks for product recovery applications are reviewed. PMID- 14538126 TI - The molecular genetics of virulence of Xanthomonas campestris. AB - Bacteria belonging to the genus Xanthomonas are important pathogens of many plants, and their virulence appears to be due primarily to secreted and surface compounds that could increase host nutrient loss, or avoid or suppress unfavorable conditions in the host. Type II and III secretory pathways are essential for virulence. Some individual extracellular enzymes (type II-secretion dependent) affect final bacterial population levels, whereas some avirulence gene products (type III-secretion dependent) affect virulence by altering host metabolism. Avr proteins, probably secreted via a pilus, can also be recognized by host resistance gene products. Virulence is also associated with bacterial surface polysaccharides, which may help to avoid host defense responses, and regulatory gene systems, which can control virulence gene expression. PMID- 14538129 TI - Microbial alkaline proteases: from a bioindustrial viewpoint. AB - Alkaline proteases are of considerable interest in view of their activity and stability at alkaline pH. This review describes the proteases that can resist extreme alkaline environments produced by a wide range of alkalophilic microorganisms. Different isolation methods are discussed which enable the screening and selection of promising organisms for industrial production. Further, strain improvement using mutagenesis and/or recombinant DNA technology can be applied to augment the efficiency of the producer strain to a commercial status. The various nutritional and environmental parameters affecting the production of alkaline proteases are delineated. The purification and properties of these proteases is discussed, and the use of alkaline proteases in diverse industrial applications is highlighted. PMID- 14538132 TI - Bioorganic reactions in microemulsions: the case of lipases. AB - Water-in-oil microemulsions, or reverse micelles, are being evaluated as a reaction medium for a variety of enzymatic reactions. These systems have many potential biotechnological applications. Important examples are the use of various lipase microemulsion systems for hydrolytic or synthetic reactions. This review illustrates the biotechnological applications of microemulsions as media for bioorganic reactions. The principal focus is on lipase catalyzed processes. PMID- 14538133 TI - Phosphate solubilizing bacteria and their role in plant growth promotion. AB - The use of phosphate solubilizing bacteria as inoculants simultaneously increases P uptake by the plant and crop yield. Strains from the genera Pseudomonas, Bacillus and Rhizobium are among the most powerful phosphate solubilizers. The principal mechanism for mineral phosphate solubilization is the production of organic acids, and acid phosphatases play a major role in the mineralization of organic phosphorous in soil. Several phosphatase-encoding genes have been cloned and characterized and a few genes involved in mineral phosphate solubilization have been isolated. Therefore, genetic manipulation of phosphate-solubilizing bacteria to improve their ability to improve plant growth may include cloning genes involved in both mineral and organic phosphate solubilization, followed by their expression in selected rhizobacterial strains. Chromosomal insertion of these genes under appropriate promoters is an interesting approach. PMID- 14538134 TI - Molecular laser biotechnology. AB - Laser technology has developed to the point where it is possible to utilize lasers as a sophisticated but accessible tool in understanding and manipulating gene functioning. This review emphasizes some of the systems that employ lasers in the new and growing field of molecular laser biotechnology. Here the main emphasis is on the manipulation and understanding of bacterial and plant systems. PMID- 14538135 TI - Emerging diseases: a global threat. AB - A growing and globalizing threat of emerging and re-emerging diseases is best addressed through reliance on rapid detection, diagnosis, and containment. The efficiency and success of this approach depends on intricate networking and collaboration among all stakeholders including intergovernmental and nongovernmental organizations and specialized agencies in the developed and developing countries. Such cooperation, while focusing on eliminating a growing threat, also helps avoid duplication of effort and improves use of scarce financial resources. This review provides a summary of the problem of emerging/re emerging diseases and the effort being directed at controlling the threat. Opportunities are identified for a more coordinated approach to addressing the problem. PMID- 14538138 TI - DNA markers in plant improvement: an overview. AB - The progress made in DNA marker technology has been tremendous and exciting. DNA markers have provided valuable tools in various analyses ranging from phylogenetic analysis to the positional cloning of genes. The development of high density molecular maps which has been facilitated by PCR-based markers, have made the mapping and tagging of almost any trait possible. Marker-assisted selection has the potential to deploy favorable gene combinations for disease control. Comparative studies between incompatible species using these markers has resulted in synteny maps which are useful not only in predicting genome organization and evolution but also have practical application in plant breeding. DNA marker technology has found application in fingerprinting genotypes, in determining seed purity, in systematic sampling of germplasm, and in phylogenetic analysis. This review discusses the use of this technology for the genetic improvement of plants. PMID- 14538139 TI - Integrated production and protection under protected cultivation in the Mediterranean region. AB - The article discusses some basic aspects of the integrated production and protection of horticultural crops, basically vegetables (e.g. tomatoes, cucumbers, peppers, eggplants, beans, etc.) and fruit trees (bananas), as well as flowers grown under cover in the Mediterranean region, namely Morocco, Syria, Lebanon, Egypt, Jordan, Tunisia, Cyprus, and Turkey. PMID- 14538140 TI - Bovine chymosin: production by rDNA technology and application in cheese manufacture. AB - Bovine chymosin, an aspartyl protease extracted from abomasum of suckling calves, is synthesized in vivo as preprochymosin and secreted as prochymosin which is autocatalytically activated to chymosin. Chymosin is bilobular, with Asp 32 and Asp 215 acting as the catalytic residues. Chymosin A and chymosin B have pH optima of 4.2 and 3.8, respectively, and act to initiate milk clotting by cleaving kappa-casein between Phe 105 and Met 106. The gene encoding chymosin has been cloned and expressed in suitable bacteria and yeast hosts under the control of lac, trp, trp-beta, gly A genes, and serine hydroxymethyl-transferase promoters. Protein engineering of chymosin has also been attempted. A number of companies are now producing recombinant chymosin for commercial use in cheese manufacture. PMID- 14538141 TI - Flow cytometry of plant cells with applications in large-scale bioprocessing. AB - In recent years, there has been a significant upsurge in the application of flow cytometry to plant cells and plant cell cultures. As well as a range of uses in plant biology, flow cytometry offers many advantages for monitoring plant cell cultures used in large-scale bioprocessing operations. This review summarizes the current status of the field, concentrating on methods for DNA measurement and multiparameter cell cycle analysis. Techniques for screening and selection of elite cell lines with high productivity of secondary metabolites are also addressed. PMID- 14538142 TI - Control of fed-batch fermentations. AB - Fed-batch fermentation is used to prevent or reduce substrate-associated growth inhibition by controlling nutrient supply. Here we review the advances in control of fed-batch fermentations. Simple exponential feeding and inferential methods are examined, as are newer methods based on fuzzy control and neural networks. Considerable interest has developed in these more advanced methods that hold promise for optimizing fed-batch techniques for complex fermentation systems. PMID- 14538143 TI - Role of micronutrients in activated sludge-based biotreatment of industrial effluents. AB - Industrial processes often produce wastewaters that resist biological treatment owing to the unfamiliarity of some components to biological systems. Availability of nutrients determines the community structure of the activated sludge and hence the efficiency of the degradation process. Micronutrients influence the bacteria involved in waste degradation and also the species diversity within the sludge. The requirements for and toxicity of different micronutrients vary according to the nature of the waste and the ecology of the sludge. Adding micronutrients to biological treatment processes is one possible approach to upgrading an existing facility in order to deal with increasing volumes and strengths of industrial wastewaters and the tightening discharge legislation. PMID- 14538144 TI - Downstream processing of algal polyunsaturated fatty acids. AB - Little information exists on recovering polyunsaturated fatty acids from microalgae; however, methods for concentration and purification of PUFAs from fish oil have been extensively reported. This review examines recovery and purification of microalgae derived PUFAs, but techniques developed for use with fish oil are also reviewed as being potentially useful for concentration and purification from microalgae. The two main techniques for concentrating and purifying-urea fractionation and high performance liquid chromatography-are discussed in depth and attention is focused on the process developed by the authors for obtaining highly pure PUFA. Other potentially useful techniques, such as supercritical fluid extraction and lipase-catalyzed processing are detailed. PMID- 14538145 TI - Probiotics: functionality and commercial status. AB - Probiotics in the form of fermented milk products have been consumed for centuries. In this century various health benefits have been purported to result from consumption of foods containing live microorganisms, particularly lactic acid bacteria (LAB). Probiotics can provide relief for lactose intolerant individuals and reduce bouts of diarrhea. Evidence for other claims such as lowering serum cholesterol, suppressing cancer and stimulating the immune system remains to be clearly established by conducting well-controlled, statistically valid clinical trials. Although the benefits to healthy individuals are uncertain, many consumers especially in Japan and Europe, perceive probiotic products to be healthful, and sales are robust. PMID- 14538148 TI - Lipase catalyzed modification of milkfat. AB - Decreasing consumption of high fat milk and dairy products is driving the dairy industry to seek other uses for increasing surplus of milkfat. Enzyme catalyzed modification of milkfat using lipases is receiving particular attention. This review examines lipase-mediated modification of milkfat. Especial attention is given to industrial applications of lipases for producing structured and modified milkfat for improved physical properties and digestibility, reduced caloric value, and flavor enhancement. Features associated with reactions such as hydrolysis, transesterification, alcoholysis and acidolysis are presented with emphasis on industrial feasibility, marketability and environmental concerns. Future prospects for enzyme catalyzed modification of milk fat are discussed. PMID- 14538149 TI - Newcastle disease vaccines. AB - Newcastle disease (ND) is a worldwide problem with severe economic implications, affecting chickens, turkeys and other birds. Newcastle disease virus (NDV), a member of the Paramyxoviridae group can cause disease of diverse severity in accordance with environmental factors. NDV strains are classified according to their virulence into three categories. The lentogenic strains are very mild and naturally inhabit healthy flocks. They can be used as live vaccines even for young chicks. Killed vaccines can be produced from the same viruses following inactivation. Mesogenic ND viruses, which cause mild or inapparent respiratory infections, have recently been banned in many countries even for killed vaccine production due to fears of disease emergence. Velogenic strains are the causative agents of the disease and can be used for the purpose of vaccine challenge test. Production and use of Newcastle disease vaccines are discussed in this review. PMID- 14538150 TI - Cell-free protein synthesis systems. AB - Cell-free protein synthesis systems enable the direct in vitro expression of proteins from template DNA or RNA. Use of biochemical and bioengineering techniques has greatly improved the yields and productivities of cell-free systems. In some cases, the yields approach the in vivo levels. Moreover, in vitro systems are capable of rapidly providing artificial polypeptides that greatly facilitate protein engineering. Post-translational modification steps in cell-free systems also offer exciting possibilities as reviewed here. PMID- 14538153 TI - Use of non-pathogenic or hypovirulent fungal strains to protect plants against closely related fungal pathogens. AB - Nonpathogenic (avirulent), or low virulent (hypovirulent) strains are capable of colonizing infection site niches on the plants' surfaces and protecting susceptible plants against their respective pathogens. Such phenomena have been demonstrated for a considerable number of plant pathogens. The modes of protection differ among the nonpathogenic strains, and one strain can protect by more than one mechanism. Competition for infection sites, or for nutrients (such as carbon, iron) as well as induction of the host plant resistance, have been demonstrated for several pathogens such as Rhizoctonia spp., Fusarium spp. and Pythium spp. Mycoparasitism was shown for Pythium spp. Transmission of double stranded RNA mycoviruses from hypovirulent strains to virulent strains renders the virulent strains hypovirulent. Chestnut trees infected with the chestnut blight pathogen, Cryphonectria (Endothia) parasitica, recovered after inoculation with transmissible hypovirulent strains. Nonpathogenic strains of various fungi are potential candidates for development of biocontrol preparations. Some strains are already used in Agriculture. PMID- 14538154 TI - Genetic engineering for cut-flower improvement. AB - The application of modern biotechnological approaches to cut flowers has clearly become instrumental and rewarding for the floriculture industry. In recent years, several gene-transfer procedures have been developed for some of the major commercial cut flowers. Using Agrobactrium or microprojectile bombardment, several basic protocols are now available. However, despite the great progress and interest in gene transfer to these crops, their transformation is routine in only a limited number of laboratories, and its application is still considered to be an "art form". This review summarizes the reported gene-transfer procedures for the main cut-flower crops, with an emphasis on the unique factors of each method and the recent progress in introducing new traits of horticultural interest into these species. PMID- 14538157 TI - Intellectual property rights and the growth of biotechnology based industries in developing countries. PMID- 14538158 TI - Cellulose degrading enzymes and their potential industrial applications. AB - Bioconversion of cellulose to soluble sugars and glucose is catalyzed by a group of enzymes called cellulases. Microorganisms including fungi, bacteria and actinomycetes produce mainly three types of cellulase components--endo-1,4-beta-D glucanase, exo-1,4-beta-D-glucanase and beta-glucosidase--either separately or in the form of a complex. Over the last several decades, cellulases have become better understood at a fundamental level; nevertheless, much remains to be learnt. The tremendous commercial potential of cellulases in a variety of applications remains the driving force for research in this area. This review summarizes the present state of knowledge on microbial cellulases and their applications. PMID- 14538159 TI - The microbial degradation of halogenated diaryl ethers. AB - The structurally related polyhalogenated diaryl ethers such as diphenyl ethers (DEs), dibenzofurans (DFs), and dibenzo-p-dioxins (DDs) are regarded, due to their physicochemical and toxicological properties, as a class of compounds giving reason for serious environmental concern. While the nonhalogenated basic structures are biodegradable under aerobic conditions, there is the need for rather specialized strains to mineralize the halogenated derivatives. Certain halogenated metabolites might cause serious problems such as having inhibitory effects upon the degradation. Anaerobic methanogenic consortia do have the ability to almost completely dehalogenate even polyhalogenated congeners. It has been shown that certain fungi are capable of transforming chlorinated DFs and DDs by the activity of nonspecific enzymes such as lignin-peroxidases. PMID- 14538160 TI - A review on disease transmission studies in relationship to production of embryos by in vitro fertilization and to related new reproductive technologies. AB - This paper addresses the circumstances of germplasm contamination and updates on transmission of pathogenic agents by embryos produced in vitro and by associated techniques. It has been shown that some pathogenic agents might have been associated with the follicular oocytes and oviductal cells, collected for in vitro fertilization (IVF), resulting in infected embryos. Experimental introduction of pathogenic agent with oocytes or infected semen into the IVF system allows, in most cases, for the fertilization of eggs and for the production of some transferable quality embryos. Rendering of oocytes and embryos free of infectious pathogens, using the standard sequential washing or enzymatic treatment, is inconsistent and more difficult in the presently used in vitro fertilization system as compared to in vivo produced embryos. PMID- 14538714 TI - The glucanases of Cellulomonas. AB - Cellulomonas is a unique bacterium possessing not only the capacity to degrade various carbohydrates, such as starch, xylan and cellulose, but crystalline cellulose as well. It has developed a complex battery of glucanases to deal with substrates possessing such extensive microheterogeneities. Some of these enzymes are multifunctional, as well as cross inducible, possessing a multi-domain structure; these enzymes are thought to have arisen by the shuffling of these domains. Intergeneric hybrids have been constructed between Cellulomonas and Zymomonas so as to enhance the industrial potential of this organism. This review examines the unique features of this microorganism and evaluates its key role in the conversion of complex wastes to useful products, by virtue of its unusual attributes. PMID- 14538715 TI - Cocoa biotechnology: status, constraints and future prospects. AB - Current status and future prospects of cocoa biotechnology are reviewed. Potential for improving and modifying cocoa bean yield and quality are discussed. Prospects for producing cocoa components in vitro and cocoa butter substitutes in crops other than cocoa are examined. Application of complementary research tools is expected to allow significant enhancements in cocoa bean yield. Furthermore, cocoa varieties with modified characteristics are likely to become available, in particular varieties with increased cocoa butter content and modified fatty acid patterns. In vitro production of cocoa components is less likely. PMID- 14538716 TI - Genetic manipulation of plant growth-promoting bacteria to enhance biocontrol of phytopathogens. AB - Plant growth-promoting bacteria (PGPB) control the damage to plants from phytopathogens by a number of different mechanisms including: outcompeting the phytopathogen, physical displacement of the phytopathogen, secretion of siderophores to prevent pathogens in the immediate vicinity from proliferating, synthesis of antibiotics, synthesis of a variety of small molecules that can inhibit phytopathogen growth, production of enzymes that inhibit the phytopathogen and stimulation of the systemic resistance of the plant. Biocontrol PGPB may be improved by genetically engineering them to overexpress one or more of these traits so that strains with several different anti-phytopathogen traits which can act synergistically are created. In engineering these strains it is essential to ensure that the normal functioning of the bacterium is not impaired, i.e., that there is no problem with metabolic load. PMID- 14539376 TI - Enzyme catalyzed esterification. AB - Enzyme catalyzed esterification reactions have found many applications, ranging from the modification of vegetable oils for human consumption to the production of optically pure chemicals. To displace the equilibrium in favor of synthesis, rather than hydrolysis, these reactions are performed in non-aqueous or microaqueous media. The influence of the amount of water, and of the nature of organic solvent, are new parameters to consider in the optimization of industrial processes. They also add a new perspective to our knowledge of the functioning of enzymes. PMID- 14539377 TI - An in vitro approach to ruminant mammary gland biology. AB - This review discusses both fundamental and applied in vitro studies on ruminant mammary gland biology and summarizes progress made over the last decade in development of in vitro techniques to study growth, function and pathology of the mammary gland. The advantages and limitations of different in vitro systems are considered including explant cultures, primary cell cultures and immortalized lines of mammary-derived cells from cow, sheep and goat. The cell growth, differentiation and response to lactogenic hormones and growth factors are discussed as well as the relevance of the cell behavior in different culture conditions. PMID- 14539378 TI - DNA based biosensors. AB - Compared to advances in enzyme sensors, immunosensors, and microbial biosensors, relatively little work exists on DNA based biosensors. Here we review the DNA based biosensors that rely on nucleic acid hybridization. Major types DNA biosensors--electrochemical, optical, acoustic, and piezoelectric--are introduced and compared. The specificity and response characteristics of DNA biosensors are discussed. Overall, a promising future is foreseen for the DNA based sensor technology. PMID- 14540156 TI - Plasmid stability in recombinant Saccharomyces cerevisiae. AB - Because of many advantages, the yeast Saccharomyces cerevisiae is increasingly being employed for expression of recombinant proteins. Usually, hybrid plasmids (shuttle vectors) are employed as carriers to introduce the foreign DNA into the yeast host. Unfortunately, the transformed host often suffers from some kind of instability, tending to lose or alter the foreign plasmid. Construction of stable plasmids, and maintenance of stable expression during extended culture, are some of the major challenges facing commercial production of recombinant proteins. This review examines the factors that affect plasmid stability at the gene, cell, and engineering levels. Strategies for overcoming plasmid loss, and the models for predicting plasmid instability, are discussed. The focus is on S. cerevisiae, but where relevant, examples from the better studied Escherichia coli system are discussed. Compared to free suspension culture, immobilization of cells is particularly effective in improving plasmid retention, hence, immobilized systems are examined in some detail. Immobilized cell systems combine high cell concentrations with enhanced productivity of the recombinant product, thereby offering a potentially attractive production method, particularly when nonselective media are used. Understanding of the stabilizing mechanisms is a prerequisite to any substantial commercial exploitation and improvement of immobilized cell systems. PMID- 14540801 TI - Microbial attachment and growth in fixed-film reactors: process startup considerations. AB - Optimal steady-state performance of any biofilm reactor requires a fully developed and mature biofilm. During fixed-film reactor startup phase, biofilm is in process of development and accordingly, process performance is difficult to quantify. Environmental, cellular and surface factors greatly influence the process of biofilm formation during reactor startup. Improved knowledge of nutritional, toxicological and environmental requirements of wastewater degrading microorganisms has helped define optimal microbial growth conditions. In case of anaerobic fixed film reactors the startup is hindered by low microbial growth rates, strict environmental requirements and limited ability of methanogens to adhere and form fixed biofilms. These obstacles could be overcome by proper support media selection and formulation of appropriate inoculation procedures and startup strategies. PMID- 14540800 TI - Biotransformation of sterols: selective cleavage of the side chain. AB - This review elaborates on the recent development of microbial sterol biotransformation systems. Particular emphasis is laid on the new enzymatic approach investigating the cleavage of sterol side chain. New developments in the area of immobilized cell system and use of organic media along with recent reviews on side chain cleavage are discussed. PMID- 14540802 TI - Aeroponics for the culture of organisms, tissues and cells. AB - Characteristics of aeroponics are discussed. Contrast is made, where appropriate, with hydroponics and aero-hydroponics as applies to research and commercial applications of nutrient mist technology. Topics include whole plants, plant tissue cultures, cell and microbial cultures, and animal tissue cultures with regard to operational considerations (moisture, temperature, minerals, gaseous atmosphere) and design of apparati. PMID- 14540890 TI - Biochemical pathways and mechanisms nitrogen, amino acid, and carbon metabolism. AB - For both nitrogen and carbon metabolism there exist specific regulatory mechanisms to enable cells to assimilate a wide variety of nitrogen and carbon sources. Superimposed are regulatory circuits, the so called nitrogen and carbon catabolite regulation, to allow for selective use of "rich" sources first and "poor" sources later. Evidence points to the importance of specific regulatory mechanisms for short term adaptations, while generalized control circuits are used for long term modulation of nitrogen and carbon metabolism. Similarly a variety of regulatory mechanisms operate in amino acid metabolism. Modulation of enzyme activity and modulation of enzyme levels are the outstanding regulatory mechanisms. In prokaryotes, attenuation and repressor/operator control are predominant, besides a so called "metabolic control" which integrates amino acid metabolism into the overall nutritional status of the cells. In eukaryotic cells compartmentation of amino acid metabolites as well as of part of the pathways becomes an additional regulatory factor; pathway specific controls seem to be rare, but a complex regulatory network, the "general control of amino acid biosynthesis", coordinates the synthesis of enzymes of a number of amino acid biosynthetic pathways. PMID- 14540891 TI - Cell growth and enzyme kinetics. PMID- 14540892 TI - Expression vehicles used in recombinant DNA technology. AB - We survey cloning vehicles whose function is to carry and express a gene in host cells including Escherichia coli, Saccharomyces cerevisiae and mammalian cells. In E. coli these include vehicles based on the lac operon, the trp operon, the rho leftward operon, and the recA gone; open reading frame cloning vehicles are also discussed, as are steps that can be taken to extrude a gene product from the cell and the use of plasmids with runaway replication. In S. cerevisiae we discuss vehicles based on the PGK gene, the ADH1 gene, the acid phosphatase gene and the GAL1-GAL10 gene cluster. In mammalian cells we discuss vehicles based on SV40 promoters, the metallothionein gene, retroviral LTR promoters, bovine papilloma virus and vaccinia virus. PMID- 14540893 TI - Process dynamics, instrumentation and control. AB - A total of 77 references pertaining to the subject were consulted to compose the present review. In addition to their years of publication (1981-1983), the references were selected primarily for their practicality and ability to provide experimental results and confirmation. First, a generalized model illustrating the biochemical reaction engineering considerations is presented. Here the knowledge of reactor and reaction kinetics is emphasized and the need for process instrumentation identified. Recent developments in key process sensors are then discussed in detail along with their utilities in various process situations. Finally, application of control principles and on-line computers to biochemical process operation is given a realistic evaluation by considering the process dynamics and instrumentation capabilities. Future challenges and opportunities from both the reaction engineering and system engineering points of view are carefully assessed. The most recent review relevant to the subject is that by Bull (1983). PMID- 14540894 TI - Downstream processing of proteins. AB - This review focuses on the recovery of proteins from fermented starting materials, covering developments that have appeared in the literature since 1979. The major subjects discussed include cell disruption and extraction, solid/liquid separations, chromatography, separations in solution, and finishing operations. While advances have occurred in every area, the greatest diversity of innovation has taken place in the fields of chromatography and tangential-flow filtration. PMID- 14540895 TI - Biotechnological applications of tissue culture to forest tree improvement. AB - Plant tissue culture techniques are of tremendous potential value to forest tree improvement. The technology is envisaged as playing a complementary role to traditional methods through exploiting spontaneous or induced genetic and epigenetic variability in culture, by use of haploidy and by the use of protoplasts. Haploids and protoplasts will aid in shortening breeding cycles and allow for unconventional crosses respectively. Clonal propagation is an integral part of any tree improvement program, and in addition can play an independent role in reforestation, clonal orchard establishment and in energy foresting. The goals, problems and limitations of these applications of tissue culture technology to forest tree improvement are indicated and assessed. PMID- 14540896 TI - Strategies for cultivation of recombinant organisms. AB - An overview on the strategies for cultivation of recombinant organisms is presented in three sections, that is, the stability of plasmids, expression of cloned genes and an example of a genetically engineered microorganism. PMID- 14540897 TI - Protoplast fusion as a means of producing new industrial yeast strains. AB - The ability of the yeast Saccharomyces cerevisiae to produce ethanol and carbon dioxide from carbohydrates has been exploited by man for thousands of years. During its brief existence protoplast fusion has already become an invaluable tool for investigating the molecular genetics of yeast, as well as an important part of the arsenal of genetic manipulations available to develop new strains. In the case of industrial strains, a mating reaction is usually lacking. Protoplast fusion overcomes this barrier and allows for the genetic analysis of commercially valuable traits. A major block toward broader applicability of fusion is that hybrids becomes more unstable as the genetic backgrounds of the parents diverge. As greater progress in overcoming this problem is made, fusion, by itself and in conjunction with classical hybridization, will become increasingly important in the development of new strains. The incorporation of cytoplasmic elements into yeast protoplasts has the potential to vastly expand the array of biochemical reactions performed by yeasts, thereby increasing the importance of this microbe to mankind. PMID- 14541212 TI - Monoclonal antibodies against bacteria. AB - Highlights are presented of most recent work in which monoclonal antibodies have been instrumental in the study of bacteria and their products. Topics summarized pertain to human and veterinary medicines, dentistry, phytopathology, ichthyology, and bacterial ecophysiology, differentiation, evolution and methanogenic biotechnology. PMID- 14541213 TI - Reversed micelles in biotechnological processes. PMID- 14541214 TI - Media for hybridoma growth and monoclonal antibody production. AB - For the economical production of monoclonal antibodies (MAbs), the cell-culture medium must be optimized for three different phases: growth of the hybridomas, MAb productivity of the hybridomas, and MAb purification or downstream processing. Medium improvements are necessary to meet these requirements for large-scale MAb production. Information bearing on this issue is being addressed in two research areas, cell biology and biochemical engineering, and is reviewed in this article. PMID- 14541215 TI - Development of recombinant Streptomyces for biotechnological and environmental uses. AB - Recombinant DNA techniques for manipulation of genes in Streptomyces are well developed, and currently there is a high level of activity among researchers interested in applying molecular cloning and protoplast fusion techniques to strain development within this commercially important group of bacteria. A number of efficient plasmid and phage vector systems are being used for the molecular cloning of genes, primarily those encoding antibiotic biosynthesis enzymes, but also for a variety of other bioactive proteins and enzymes of known or potential commercial value. In addition, cloning aimed at constructing specialized bioconversion strains for use in the production of chemicals from organic carbon substrates is underway in numerous laboratories. This review discusses the current status of research involving recombinant DNA technologies applied to biotechnological applications using Streptomyces. The topic of potential environmental uses of recombinant Streptomyces is also reviewed, as is the status of current research aimed at assessing the fate and effects of recombinant Streptomyces in the environment. Also summarized is recent research that has confirmed that genetic exchange occurs readily among Streptomyces in the soil environment and which has shown the potential for exchange between recombinant Streptomyces and native soil bacteria. PMID- 14541216 TI - Biologicals produced from animal cells in culture--an overview. AB - A wide spectrum of biologicals are produced from animal cells in culture. Among these biologicals are viral vaccines (human and veterinary), monoclonal antibodies, immunoregulators, enzymes, hormones, polypeptide growth factors, viral bioinsecticides, tumor antigens, cell mass as a product and reconstitution of living skin. For research and development (R&D) and production of these products requires knowledge and experience in one or more of the following advanced technologies: 1) Development of novel as well as modification of conventional cultivation equipment; 2) Adaptation of cultivation techniques to the production of the desired product; 3) Hybridization technologies; 4) Genetic engineering techniques suitable for animal cells; 5) Development of a wide range of microcarriers and fixed-bed culturing systems; 6) Microencapsulation techniques; 7) Development of suitable media for cell cultivation; 8) Adaptation of suitable protein concentration and purification techniques. PMID- 14541775 TI - Mass transfer limitations in immobilized cells. PMID- 14541776 TI - Acetone-butanol fermentation and its variants. AB - Recent intensive research on the acetone-butanol-ethanol and the isopropanol butanol-ethanol fermentation has increased the basic understanding of these processes substantially. Metabolic investigations on Clostridium acetobutylicum, and Clostridium beijerinkii show that enzyme activities necessary for solvent production are induced only in solvent-producing cells. Although produced, or added, acetic and butyric acid have significant effects on the metabolic activities, the transition from acid to solvent production cannot as yet be fully explained. Based on studies in continuous cultures, the kinetics of product formation can be described. Knowledge of the mechanism of butanol toxicity is accumulating but no dramatic increase in butanol tolerance has so far been obtained. Successful results, approaching the limitations determined by biological and technological possibilities, have been obtained in batch and continuous cultures, and in continuous processes based on immobilized cells. Continuous processes are superior to batch cultures in respect of their productivity. PMID- 14541777 TI - Biotechnological applications of plant cells in culture. AB - For many workers, the most exciting recent advances in the realm of plant cell biotechnology, center on results obtained from experiments concerned with the genetic engineering of plant cells. Various groups of workers have managed to introduce new genetic material into plant cells, using Ti-plasmids (or modified Ti-plasmids) from Agrobacterium tumefaciens. This genetic material has been expressed (with varying degrees of efficiency), in each case. Thus the way may possibly be coming clear to produce plant cell cultures, or whole plants with entirely new or novel properties. Other areas in which progress has been made, are in the design of media conditions to promote secondary product formation, and in ways of immobilizing plant cells and enzymes, to achieve efficient secondary product formation. PMID- 14541778 TI - Fuels and industrial chemicals through biotechnology. 2. AB - An overview of research on the biotechnical production of fuels and industrial chemicals during the two-year period of 1983-1984 is presented. Ethanol fermentation has continued to be the subject of major interest. A considerable amount of work has been directed to alternative feedstocks such as pentose sugars and lactose, and to bacterial fermentations. Reports on extrusion cooking as a continuous pretreatment method for subsequent ethanol fermentation, and on novel alternative downstream processing techniques have been published. In addition to ethanol fermentation, much attention has been paid to the biotechnical production of 2,3-butanediol, and of a number of organic and amino acids. In general, there appears to be a growing interest in the application of biocatalysis for the production of specialty chemicals, although only a few examples will be discussed in this paper. The construction of a demonstration plant to produce ethanol from molasses by a two 10 kL bed-volume immobilized yeast bioreactors at the Kyowa Hakko Kogyo Company Hofu plant, the announcement by Nitto Chemical Industries Company to begin the biotechnical production of acrylamide, and the French decision to construct pilot plants for the biotechnical production of acetone butanol-ethanol cosolvent and of ethanol from renewable resources represent major scale-up developments. PMID- 14541779 TI - Bio-surfactants. AB - Interest in microbially produced biosurfactants has increased recently, due mainly to their potential as agents in enhanced oil recovery. A variety of microbes and their products have been assessed for their surface-active properties, and it has been suggested that biosurfactants may prove useful in a broad spectrum of potential applications which presently utilise synthetic surfactants. The most commonly produced biosurfactants tend to be glycolipids, usually a mono- or di-saccharide attached to a fatty acid, but more complex molecules such as lipopeptides, lipoproteins, and lipo-heteropoly-saccharides have been isolated and studied. Biosurfactant production by microbes is often but not invariably enhanced by the addition of hydrocarbon to the growth medium, and needs to be optimised by controlling such factors as carbon source, nitrogen source and concentrations, aeration and metal ions. Biosurfactants have been shown to be as effective, if not more so, than many conventional synthetic surfactants and their future utilisation may depend utilimately upon the prevailing economics for their production. PMID- 14542050 TI - On the merits of viable-cell immobilisation. AB - Many advantages have been claimed over the years for the use of immobilised cells, both as enzyme systems and as whole viable cell systems for complete fermentation reactions. However, few of the claims have been fully substantiated, and may not even be entirely justified. Most research is involved with single applications and the best that can be hoped for is some evidence that immobilised cells in each of these individual cases display some advantage over the equivalent free cell system. The purpose of this review is to assess the merits of viable cell immobilisation in the light of published literature and to elucidate the underlying mechanisms. Particular attention is paid to the generally unanticipated, but widely observed enhanced stability of immobilised cell fermentation processes. PMID- 14542051 TI - Microbial agents for decolorization of dye wastewater. AB - Colored dye wastewater presents a formidable task for biological treatment. Depending on how it is generated, wide pH spans and high salt concentrations such as chloride ion often add to the difficulties. Systematic screening for dye decolorizing and/or degrading bioagents from soil and water samples discovered fungi which show dramatic color removal capability (Shen, et al., 1990). One example shows that up to 99% reduction of light absorption at characteristic wavelength of a red dye (200 mg/L) could be obtained within 48 hours. This ability does not appear to be specific toward dyes targeted for action. It clarifies, often beyond detection by naked eyes, a repertoire of colored wastewater samples. These results appeared to be insensitive to wide variations in pH and salt concentration and, they are not limited to one particular fungal species or genus either upon further investigation. This dye adsorption mechanism may be of great significance in uncovering new methods for bio-removal or bio recovery of dye substances in wastewater. PMID- 14542052 TI - Regulation of ACV synthetase in penicillin- and cephalosporin-producing microorganisms. AB - ACV synthetase is the first enzyme in the biosynthetic pathway for all natural penicillins and cephalosporins. Its activity catalyzes the possible rate-limiting step and is subject to various regulatory controls. In both the fungus Cephalosporium acremonium and the actinomycete Streptomyces clavuligerus, formation of the enzyme is repressed by ammonium and phosphate ions, but not by easily-utilized carbon sources; it is induced by methionine in C. acremonium. The action of the crude enzyme is indirectly inhibited in vitro by sugars such as glucose and by the carbon source metabolite glyceraldehyde-3-phosphate (G3P). Sugars are not inhibitory to the purified enzyme activity but G3P is inhibitory. The sugar inhibition is reversed by ATP and the G3P inhibition by L-cysteine (L cys). Addition of L-cys to fermentation media increases beta-lactam production by both microorganisms. Phosphate and ferrous ions inhibit enzyme activity. Dissolved oxygen levels do not affect enzyme formation. Regulation of ACVS formation most likely occurs at the transcriptional level. PMID- 14542053 TI - Industrial enzymes--developments in production and application. AB - The utilization of gene technology and of new production technologies have made industrial enzymes with improved properties or better cost performance available. This has in turn opened important new areas of enzyme applications. The benefits to the customers are considerable: cost savings in the application process, improved product quality, and in most cases also a significantly reduced impact on the environment. Gene technology offers several benefits to the enzyme industry. This technology enables the use of safe, well-documented host organisms easy to cultivate, the microbial production of enzymes of animal and plant origin, the realization of enhanced efficiency and high product purity, and also the production of enzymes with improved stability and activity. Developments in production technology include advanced control methods, the use of expert systems, and the application of large-scale crystallization. As case stories the development of a lipase and of a cellulase is described. The effect on environment of enzyme application and production is discussed. PMID- 14542171 TI - Enhancement of oxygen transfer rates in fermentation using oxygen-vectors. AB - Oxygen transfer is one of the bottlenecks in conventional fermentation technology and it has so far been almost totally overlooked with regards to high cell densities and immobilized cells. This review presents some new concepts to improve oxygen supply in aerobic fermentations, especially the use of oxygen vectors. The oxygen-vectors generally used are liquids which are insoluble in the fermentation media. Their utilization in an emulsified form can significantly increase the oxygen transfer coefficient between gas and aqueous phases. It seems that the vector acts as an active intermediate in the oxygen transport from gas bubbles to aqueous phase, but the mechanisms involved in this unconventional technique of aeration are not yet known. PMID- 14542172 TI - Hydrophobic interaction chromatography. AB - Hydrophobic interaction chromatography (HIC) is emerging as a useful technique for the separation of biological compounds. Advances in the past two years in HIC applications, stationary phases, eluents, and theory are reviewed. Recent applications of HIC processes include analytical and semi-preparative separations of a variety of proteins, such as isolectins, hemoglobins, calmodulin, and cardiotoxins. Additionally, HIC is being employed as a tool to investigate protein properties and mechanisms. Advances in HIC stationary phases include development of non-porous, microparticulate supports as well as supports with pore sizes up to 1000 Angstroms. Studies of HIC eluents have further shown the effects of mobile phase pH, water-structuring characterization, and surface tension increments on retention. Various retention mechanisms which have been presented are reviewed; and a correlation relating resolution to column and solute parameters is presented. Protein conformational effects at specific sites have been shown to have a significant impact on retention and specific examples illustrating such effects are discussed. PMID- 14542173 TI - Molecular genetics of Rhizobium Meliloti symbiotic nitrogen fixation. AB - The application of recombinant DNA techniques to the study of symbiotic nitrogen fixation has yielded a growing list of Rhizobium meliloti genes involved in the processes of nodulation, infection thread formation and nitrogenase activity in nodules on the roots of the host plant, Medicago sativa (alfalfa). Interaction with the plant is initiated by genes encoding sensing and motility systems by which the bacteria recognizes and approaches the root. Signal molecules, such as flavonoids, mediate a complex interplay of bacterial and plant nodulation genes leading to entry of the bacteria through a root hair. As the nodule develops, the bacteria proceed inward towards the cortex within infection threads, the formation of which depends on bacterial genes involved in polysaccharide synthesis. Within the cortex, the bacteria enter host cells and differentiate into forms known as bacteroids. Genes which encode and regulate nitrogenase enzyme are expressed in the mature nodule, together with other genes required for import and metabolism of carbon and energy sources offered by the plant. PMID- 14542174 TI - Use of Xenopus oocytes to study the expression of cloned genes and translation of mRNA. AB - One of the most active areas of research in the field of molecular biology is the examination of the mechanisms associated with the regulation of gene expression. Our understanding of the events in eukaryotic transcription has been aided by the ability to test the expression of various genomic DNA constructs after their microinjection into the germinal vesicle of Xenopus oocytes. This in vivo transcription system has permitted the analysis of the involvement of cis-acting DNA sequences and the examination of the effect of co-injected trans-acting factors on gene expression. Furthermore, the Xenopus oocyte has been employed widely as an in vivo translation system. Not only is the Xenopus oocyte system a sensitive assay for the translation of rare mRNAs but it also has the ability to post-translationally modify and compartmentalize numerous types of proteins. Finally, the Xenopus oocyte has proven useful in the procedures associated with the cloning and screening of cDNAs. PMID- 14542393 TI - Fed-batch culture technology. AB - The use of fed-batch procedures offers distinct advantages over other modes of operation of bioreactors, and is a widely researched technique. These advantages are discussed; some uses of fed-batch procedures and the associated methods of modelling and control are reviewed. PMID- 14542394 TI - Fermented foods, feeds, and beverages. AB - World population reached 5 billion people in 1986 and is expected to rise to 6 particularly severe in parts of Africa, particularly Ethiopia. Hunger and malnutrition accompany the poverty that is characteristic of the masses of the people in the developing world. Even in the United States, one of the most affluent countries, there has been a considerable increase in the number of homeless people including mothers and children living in the streets or in substandard housing. The indigenous fermented foods and beverages already consumed for centuries by hundreds of millions of the world's masses can be used in many cases to improve and extend the world's food and nutrition supply at a relatively low cost. Fermented foods, feeds and beverages are getting ever increasing attention particularly in the developing world and also in the developed world. This paper summarizes activities and advances related to fermented foods, feeds, and beverages over the past several years. PMID- 14542395 TI - Microbial polysaccharides with actual potential industrial applications. AB - The microbial polysaccharides reviewed include xanthan gum, scleroglucan, PS-10, PS-21 and PS-53 gums, polysaccharides from Alcaligenes sp., PS-7 gum, gellan gum, curdlan, bacterial alginate, dextran, pullulan, Baker's Yeast Glycan, 6-deoxy hexose-containing polysaccharides and bacterial cellulose. Factors limiting the commercial potential of certain microbial polysaccharides such as availability, rheological properties, and polyvalency are outlined. The polysaccharides are classified according to their uses as viscosity-increasing agents and as gelling agents. A third category includes polysaccharides with specific applications such as tailor-made dextran and pullulan and polysaccharides used as substrates for the preparation of rare sugars. The difficulties encountered in development of a polysaccharide at the industrial level are pointed out. PMID- 14542396 TI - Environmental implications of recombinant DNA technology. AB - Applications of recombinant DNA technology are discussed as a backdrop for evaluation of the environmental impacts of this technology. Some of applications include using traditional biological techniques for specific purposes, including nitrogen fixation, microbial pesticides, and waste treatment. In these instances the final product lies along a continuum, beginning with an organism marginally performing its function, and ending with one that is highly specialised and very efficient in what it does. One may move along this continuum toward the 'perfect' microorganism by using traditional methodologies of mutagenesis and selection, recombinant DNA technology, or a combination of the two. PMID- 14542397 TI - Production of plant secondary metabolites without plants: a perspective. AB - Efforts to commercially exploit native plant secondary metabolite production patterns in cell culture systems have been largely thwarted by the repression of secondary metabolism under growth-oriented culture conditions. Where expression can be obtained by selection or elicitation, the difficulties of large scale cultivation/processing still make the cost effectiveness of cell culture systems dubious except where a very high value market niche can be identified. The long range prospect for efficiently utilizing the catalytic genius of higher plants resides in the transfer of appropriate genetic information to microbial systems for whom the fermentation expertise and industrial facilities already exist. PMID- 14542820 TI - Inducing malolactic fermentation in wines. AB - Malolactic fermentation (MLF) in wine can be accomplished by relying on the natural microflora or by inducing through inoculation of a specific strain(s) of malolactic bacteria, primarily strains of Leuconostoc oenos. Problems with inducing MLF include intrinsic factors of the grape must such as pH, presence of sulfur dioxide, and ethanol in addition to antagonism of malolactic bacteria by wine yeast. Current methods and new technology to improve the predictability of MLF are discussed. PMID- 14542821 TI - Isolation, characterization and manipulation of cellulase genes. AB - The complete hydrolysis of cellulose requires a number of different enzymes including endoglucanase, exoglucanase and beta-glucosidase. These enzymes function in concert as part of a 'cellulase'complex called a cellulosome. In order (i) to develop a better understanding of the biochemical nature of the cellulase complex as well as the genetic regulation of its integral components and (ii) to utilize cellulases either as purified enzymes or as part of an engineered organism for a variety of purposes, researchers have, as a first step, used recombinant DNA technology to isolate the genes for these enzymes from a variety of organisms. This review provides some perspective on the current status of the isolation, characterization and manipulation of cellulase genes and specifically discusses (i) strategies for the isolation of endoglucanase, exoglucanase and beta-glucosidase genes; (ii) DNA sequence characterization of the cellulase genes and their accompanying regulatory elements; (iii) the expression of cellulase genes in heterologous host organisms and (iv) some of the proposed uses for isolated cellulase genes. PMID- 14542822 TI - Preclinical and pharmacology and toxicology of hematopoietic growth factors. AB - Initial evaluation of the safety of biological products, whether manufactured by biotechnology or not, is dictated by the extent of knowledge of the biological effects in vitro, in animals, and in man at the proposed dose and duration of administration. The quantity of useful information that will be gained from an animal study is considered for each product on a case-by-case basis. This requires consideration of the availability of a relevant species. Relevance is determined by presence and relative affinity and distribution of receptors for the biological product, relative potency in induction of biological activity related to the putative mechanism of action and pharmacokinetic considerations. The impurities and immunogenic potential of the product are also considered in designing the preclinical studies. After evaluating all these factors, the residual lack of knowledge is FDA's basis for recommending specific preclinical testing. The considerations inherent in the preclinical evaluation of factors acting primarily on hematopoietic cells, erythropoietin (EPO), interleukins (IL's) and colony stimulating factors (CSF) (1) will be discussed using case examples. PMID- 14542823 TI - Monoclonal antibody technology for mycotoxins. AB - Specific monoclonal antibodies (MABs) against aflatoxins, ochratoxin A, zearalenone, diacetoxyscirpenol and T-2 toxin have been prepared in various laboratories by the application of hybridoma technology to mycotoxins. These antibodies can be selected for sensitivity, reduced cross-reactivity, reliability and ease of production. When a suitable antibody is chosen it can then be used in a rapid immunological method such as an enzyme-linked or radio-immunoassay or immunoaffinity chromatography system. These assays have a lower limit of mycotoxin detection in the ng/ml range and have been applied to the determination of mycotoxins in samples such as maize, peanuts, peanut butter, milk and porcine kidneys. Using these immunoassay techniques, sample preparation has generally been simplified to a matter of solvent extraction of mycotoxins from the sample followed by dilution; under these conditions, levels of 1-5ug of mycotoxins/kg of sample can be found. The application and advantages of MABs to mycotoxins and the use of these antibodies in various assay techniques is discussed. PMID- 14542986 TI - Downstream processing of proteins: recent advances. AB - This review on the downstream processing of proteins describes innovations that have occurred in the field since 1983. Several areas have seen particularly high levels of achievement, and are accorded expanded coverage relative to our previous review [1]. As an example, the increasing integration of downstream operations with upstream technologies, such as molecular biology and fermentation, has led to the development of some very powerful processes. The degree to which organizations understand that there needs to be one unified process, rather than the independent steps of cloning, fermentation and recovery, seems directly related to the ultimate speed and success of the development effort. In 1983 one of the most active development areas was chromatography, especially affinity chromatography. This is still true today, and this topic has been expanded to include biospecific adsorptions that would not traditionally be classified as chromatography. With more proteins being developed for human administration, there has been an increased emphasis on all aspects of process hygiene. In addition, there has been much discussion about the impact of regulatory demands on the design and development of the manufacturing processes. Therefore, a section has been added which covers several of the regulatory issues that have been raised for products of the new biotechnology. Finally, as some of the early process development achievements are now beginning to bear fruit in the form of patents, we have increased our citation of this area of the literature. PMID- 14542987 TI - Use of cowpea trypsin inhibitor (CpTI) to protect plants against insect predation. AB - Transgenic tobacco plants expressing a cowpea trypsin inhibitor gene have enhanced levels of insect resistance to a variety of insect pests. Furthermore, insect bioassay has shown the cowpea trypsin inhibitor to have anti-metabolic activity to insect pests of the orders Lepidoptera, Coleoptera and Orthoptera. The advantages and disadvantages of this approach to developing insect resistant crops is discussed in relationship to other methods, including conventional plant breeding and chemical control. Eventually it is hoped that African farmers will benefit from this industrially sponsored research. PMID- 14542988 TI - Resource and energy recovery options for fermentation industry residuals. AB - Over the last 40 years, the fermentation industry has provided facility planners, plant operators and environmental engineers with a wide range of residuals management challenges and resource/energy recovery opportunities. In response, the industry has helped pioneer the use of a number of innovative resource and energy recovery technologies. Production of animal feed supplements, composts, fertilizers, soil amendments, commercial baking additives and microbial protein materials have all been detailed in the literature. In many such cases, recovery of by-products significantly reduces the need for treatment and disposal facilities. Stable, reliable anaerobic biological treatment processes have also been developed to recovery significant amounts of energy in the form of methane gas. Alternatively, dewatered or condensed organic fermentation industry residuals have been used as fuels for incineration-based energy recovery systems. The sale or use of recovered by-products and/or energy can be used to offset required processing costs and provide a technically and environmentally viable alternative to traditional treatment and disposal strategies. This review examines resource recovery options currently used or proposed for fermentation industry residuals and the conditions necessary for their successful application. PMID- 14542989 TI - Substrate inactivation of enzymes in vitro and in vivo. AB - Some enzymes are inactivated by their natural substrates during catalytic turnover, limiting the ultimate extent of reaction. These enzymes can be separated into three broad classes, depending on the mechanism of the inactivation process. The first type is enzymes which use molecular oxygen as a substrate. The second type is inactivated by hydrogen peroxide, which is present either as a substrate or a product, and are stabilized by high catalase activity. The oxidation of both types of enzymes shares common features with oxidation of other enzymes and proteins. The third type of enzyme is inactivated by non oxidative processes, mainly reversible loss of cofactors or attached groups. Sub classes are defined within each broad classification based on kinetics and stoichiometry. Reaction-inactivation is in part a regulatory mechanism in vivo, because specific proteolytic systems give rapid turnover of such labelled enzymes. The methods for enhancing the stability of these enzymes under reaction conditions depends on the enzyme type. The kinetics of these inactivation reactions can be used to optimize bioreactor design and operation. PMID- 14543142 TI - Microbial desulphurization of heavy oils and bitumen. AB - Most oil producing countries have extensive reserves of heavy oil and bitumen. As easily accessible sources of conventional crudes decline, these reserves will become more important in supplementing the energy requirements. Heavy oil and bitumen are highly viscous and contain 3 to 6% sulphur. These objectionable quantities of sulphur must be removed before being acceptable as refinery feedstock. This paper addresses the potential of biological desulphurization of heavy oil and bitumen. The aerobic and anaerobic processes to remove organic as well as inorganic sulphur have been reviewed. To date, most studies were performed with model substrates, particularly dibenzothiophene (DBT) in a synthetic medium. Early work concerned with the isolation of microorganisms, identification and characterization of intermediate metabolites, and the development of growth media. No commercially viable process has emerged since the engineering details of the process have not been addressed conclusively. Due to high utility and catalyst cost conventional hydrodesulphurization processes are reported to be uneconomic in case of high sulphur oils. Microbial desulphurization, on the other hand, appears to be promising due to the inherent low energy requirement. This process may become more attractive by the application of genetically modified bacteria and improvements in bioreactor design. PMID- 14543143 TI - Advances in the molecular biology of plant seed storage proteins. AB - Plant seed storage proteins were among the first proteins to be isolated (20); however, only recently, as a result of using molecular biology techniques, have the amino acid sequences of many of these proteins been determined. With the accumulation of amino acid sequence data for many vicilin-type storage proteins much has been learned concerning the location of conserved amino acid regions and other regions which can tolerate amino acid sequence variation. Combining this knowledge with recent advances in plant gene transfer technologies will allow molecular biologists to correct (by using amino acid replacement mutations) the sulfur amino acid deficiency inherent to bean seed storage proteins. The development of more nutritious soybean and common bean seeds will be of benefit to programs involving human and animal nutrition. PMID- 14543144 TI - Biocatalysis in multi-phase reaction mixtures containing organic liquids. AB - A wide range of enzymes and whole microbial cells will act as catalysts in reaction mixtures that contain 2 or more phases, one of which is an organic liquid (either a reactant or including water-immiscible organic solvents). These "biphasic" systems have a variety of structures, knowledge of which aids predictions about biocatalyst activity and stability. There is often a dilute aqueous solution phase (containing the biocatalyst), which may be emulsified with the organic phase, or "trapped" within catalyst particles; sometimes however there may only be traces of water adsorbed to the enzyme or cells. These reaction systems offer several advantages for industrial applications, notably the higher solubilities of many reactants of interest, and the ability of readily available hydrolytic enzymes to catalyse syntheses. The most non-polar organic liquids are least likely to inactivate biocatalysts, though many do remain active with relatively polar solvents. Modification of the biocatalyst may stabilise against inactivation, especially where this is due to direct contact with the phase interface. The mass transfer processes required in these systems remain poorly understood, particularly because the interfacial area is often unknown. Attractive continuous reactors may be operated using a packed bed of catalyst with a trapped aqueous phase. PMID- 14543145 TI - Microbial degradation of synthetic recalcitrant compounds. AB - Synthetic compounds, particularly highly chlorinated aromatics, comprise the bulk of the environmental pollutants that somehow must be removed from the environment. Microbial degradation of such compounds is usually very slow, making them highly persistent in nature. Some synthetic compounds, with a lower degree of chlorination are, however, biodegradable; biochemical, genetic, and molecular studies demonstrate the evolution of new plasmid-encoded enzymatic activities specifically designed for the chlorinated substrates. Nucleotide sequences of many of the genes encoding such enzymatic activities demonstrate considerable homology either near the active sites or throughout the molecules with the chromosomal genes encoding enzymes catalyzing analogous reactions. In some cases, unique repeated sequences, reminiscent of prokaryotic insertion sequence elements, are present at or near the newly evolved genes. This suggests gene duplication and divergence as well as recombinational events mediated by transposable type elements as key ingredients in the evolution of new degradative functions. An understanding of such evolutionary processes is an essential feature for the development of genetically-improved bacteria capable of utilizing and thereby removing highly chlorinated environmental pollutants from our environment. PMID- 14543146 TI - Metal resistance and accumulation in bacteria. AB - Recent research on the ecology, physiology and genetics of metal resistance and accumulation in bacteria has significantly increased the basic understanding of microbiology in these areas. Research has clearly demonstrated the versatility of bacteria to cope with toxic metal ions. For example, certain strains of bacteria can efficiently efflux toxic ions such as cadmium, that normally exert an inhibitory effect on bacteria. Some bacteria such as Escherichia coli and Staphylococcus sp. can volatilize mercury via enzymatic transformations. It is also noteworthy that many of these resistance mechanisms are encoded on plasmids or transposons. By expanding the knowledge on metal-resistance and accumulation mechanisms in bacteria, it may be possible to utilize certain strains to recover precious metals such as gold and silver, or alternatively remove toxic metal ions from environments or products where their presence is undesirable. PMID- 14543147 TI - Membrane bioreactors: Engineering aspects. AB - Membrane bioreactors have in-situ separation capability lacking in other types of immobilized cell reactors. This makes them very useful for certain systems. Enzyme reactions utilizing cofactors and hydrolysis of macromolecules are advantageous in membrane reactors. Anaerobic cell culture may be efficiently carried out in membrane cell recycle systems, while aerobic cultures work well in dual hollow fiber reactors. Animal and plant cells have much a better chance of success in membrane reactors because of the protective environment of the reactor and the small oxygen uptake rate of these cells. PMID- 14543438 TI - Use of biotechnology in mining and metallurgy. AB - Biotechnology continued to gain importance in the mineral industry during the past four years. This upsurge of interest is especially expressed in the areas of biodesulfurization of coal, recovery of precious metals from pyrite- and arsenopyrite- containing minerals, biosorption processes and biogenetic engineering. PMID- 14543439 TI - Stress inducing factors of strains of Rhizobium phaseoli as related to inoculant carrier preparations. PMID- 14543440 TI - Approach to maceration mechanism in enzymatic pulping of bast fibers by alkalophilic pectinolytic enzymes produced by Erwinia species. AB - Tissue maceration was generally elucidated by the action of endo polygalacturonase and endo-pectate or -pectin lyase (endo-PAL or -PNL). In a process of screening of Erwinia and Pseudomonas strains for enzymatic pulping of pectocellulosic bast fibers, it was found that their PAL productivity was not completely related with defibration activity, i.e., the fact that an E.chrysanthemi strain showed high PAL productivity but possessed rather low defibration activity. Moreover, defibration activity was parallel to the amount of neutral sugars released during pulping. Based on these fact, the maceration or enzymatic pulping of basts was estimated to proceed not only by cleavage of interfiber bonding cause by PAL action but also another factors. Among three possibilities proposed on the maceration mechanism of basts, it was elucidated by a concerted action of PAL and PNL with an aid of xylanase. In addition, a quantitative determinative method of maceration activity toward basts was also presented. PMID- 14543441 TI - Thermostable enzymes. AB - In general, enzyme thermostability is an intrinsic property, determined by the primary structure of the protein. However, external environmental factors including cations, substrates, co-enzymes, modulators, polyols and proteins often increase enzyme thermostability. With some exceptions, enzymes present in thermophiles are more stable than their mesophilic counterparts. Some organisms produce enzymes with different thermal stability properties when grown at lower and higher temperatures. There are commercial advantages in carrying out enzymic reactions at higher temperatures. Some industrial enzymes exhibit high thermostability. More stable forms of other industrial enzymes are eagerly being sought. PMID- 14543686 TI - Enzyme deactivation. AB - Enzyme deactivation kinetics is often first-order. Different examples of first order deactivation kinetics exhibited by different enzymes under a wide variety of conditions are presented. Examples of both soluble and immobilized enzymes are presented. The influence of different parameters, chemical modification of specific residues, inhibitors, inactivators, protecting agents, induced conformational changes by external agents, enzyme concentration, and different substrates on the first-order inactivation kinetics of different enzymes is analyzed. The different examples presented from a variety of different areas provides a judicious framework and collection demonstrating the wide applicability of first-order deactivation kinetics. Examples of reversible first order deactivation kinetics and deactivation-disguise kinetics are also presented. Different mechanisms are also presented to model complex enzyme deactivations. The non-series type mechanisms are emphasized and these involve the substrate and chemical modifiers. Substrate-dependent deactivation rate expressions that are of "separable" and "non-separable" type are presented. Rate expressions involving time-dependent rate constants along with their corresponding mechanisms are presented. Examples of enzymes that exhibit a deactivation-free grace period are also given. An interesting case of enzyme inactivation is the loss of activity in the presence of an auto-decaying reagent. The method is presented by which the intrinsic inactivation rate constants may be obtained. Examples of pH-dependent enzyme inactivation are presented that may be modelled by a five-step (or a simplified two-step) mechanism, and also by a single-step mechanism involving residual activity for the final state. Appropriate examples of enzyme inactivation are presented in each case to highlight the different mechanisms involved. PMID- 14543690 TI - Regulation of D-xylose utilization by hexoses in pentose-fermenting yeasts. AB - The aldopentose D-xylose is one of the most abundant sugars in plant biomass and its efficient microbial utilization is of fundamental importance in the overall bioconversion of lignocellulosic materials into liquid fuels and chemicals. The discovery of pentose-fermenting yeasts in the early 1980's led to world wide interest because of the perceived potential for improved D-xylose fermentation to enhance the prospect of biomass conversions. However, the utilization of D-xylose by pentose-fermenting yeasts can be adversely affected by the hexoses, mainly D glucose and D-mannose, which are usually present in high amounts in lignocellulosic hydrolysates. Research in the past several years has uncovered some of the regulatory effects of D-glucose on D-xylose utilization. However, much remains unknown about the mechanisms responsible for these effects. This review summarizes the current state of knowledge on the induction, repression and inactivation of D-xylose utilization in pentose-fermenting yeasts. PMID- 14543691 TI - Large scale protein separations: engineering aspects of chromatography. AB - The engineering considerations common to large scale chromatographic purification of proteins are reviewed. A discussion of the industrial chromatography fundamentals is followed by aspects which affect the scale of separation. The separation column geometry, the effect of the main operational parameters on separation performance, and the physical characteristics of column packing are treated. Throughout, the emphasis is on ion exchange and size exclusion techniques which together constitute the major portion of commercial chromatographic protein purifications. In all cases, the state of current technology is examined and areas in need of further development are noted. The physico-chemical advances now underway in chromatographic separation of biopolymers would ensure a substantially enhanced role for these techniques in industrial production of products of new biotechnology. PMID- 14543692 TI - Recent advances in microalgal biotechnology. AB - In the last ten years, several reviews (Benemann et al, 1987; Goldman, 1979a; Richmond, 1986b; Soeder, 1980) and books (Borowitzka and Borowitzka, 1988b; Richmond, 1986a; Lembi and Waaland, 1988) have been published on the historical background of mass cultivation of microalgal biomass, and its possible commercial applications. This review presents a brief description of the concept of microalgal biotechnology, and describes some of the recent developments, mainly in the application and commercial development of this relatively new biotechnology, Finally, an attempt is made to indicate those areas where current research and development are paving the way for future applications. PMID- 14543693 TI - Manufacture of biopharmaceutical proteins by mammalian cell culture systems. AB - In the last several years, dramatic advances have been in the development of new biopharmaceuticals including monoclonal antibodies for diagnosis and treatment and such genetically engineered proteins as tPA, Factor VIIIc, erythropoietin and soluble CD4, an anti-AIDS protein. Currently, there are several hundred such candidate drugs in human clinical trials. In most cases, these protein-based drugs will require manufacture by mammalian cell culture due to the inability of lower organisms to properly glycosylate, fold, make correct disulfide bonds and secrete active biomolecular forms. The need for large scale production from cell culture will greatly increase as more of the products in clinical trials are approved for commercial production. This will require significant reduction in manufacturing costs per gram, concomitant with increased capacity to hundreds or perhaps even thousands of kilograms annually. As an example, Invitron's multi reactor manufacturing facility has operated at greater than one-half million liters per year and has experience with more than 250 mammalian cell lines for producing protein drug products. PMID- 14543694 TI - An overview of continuous protein purification processes. AB - As the sphere of influence of recombinant technology moves away from the laboratory bench, towards product commercialization, development of manufacturing and large scale process technology is becoming a major challenge and determinant for commercial success. The challenge is particularly acute for protein purification process development where protein purification costs tend to dominate overall process economics. The primary objective for process scale purification is to minimize cost for a purified product which meets specifications. Continuous processes may be used to facilitate achievement of the overall objectives. This review critically examines the use of continuous processing for protein purification and recovery operations. The processes have been divided into three general areas: adsorptive and chromatographic, electrophoretic, and extractive. Consideration is given to the operational advantages and limitations of the reviewed processes. PMID- 14543695 TI - Immobilized mammalian cell cultivation in hollow fiber bioreactors. AB - Cultivation of animal cells for the production of recombinant proteins is an important method for manufacturing complex proteins requiring posttranslational processing. One of the often considered methods for cultivation is by immobilization of the cells in hollow fiber bioreactors (HFBRs). These systems allow the cells to grow to high densities in a shear protected environment; furthermore the product can be accumulated in high concentration in the case of ultrafiltration HFBRs. Operation and scale-up are constrained by nutrient and product transport with oxygen transfer to growing cells being the most critical parameter. Mathematical models describing HFBRs have proved to be useful in quantitating and understanding the constraints and guiding the scale-up of this approach to animal cell cultivation. PMID- 14543704 TI - Polymerase chain reaction (PCR) techniques for site-directed mutagenesis. AB - Polymerase chain reaction (PCR) technology has revolutionized the process of isolating and amplifying segments of DNA. One powerful application of PCR is its use in precise site-directed mutagenesis (SDM). SDM provides an elegant tool for scientists and engineers to explore biocatalytic mechanisms and processes to understand the structural-functional relationships of enzymes and other proteins. This article reviews techniques and methodology used in site-directed mutagenesis of genes by PCR. PMID- 14543705 TI - Bioaugmentation for enhancing biological wastewater treatment. AB - The literature on bioaugmentation products has been reviewed. Their manufacture and method of use is explained. The various applications are listed and the independent investigations, as opposed to manufacturers accounts, at laboratory and full scale are reviewed. The economics and kinetic modelling are also discussed. In laboratory investigations bioaugmentation often failed, whereas at full scale it was often successful, probably due to the imposition of steady state at laboratory scale. Most products require a period of acclimatisation before working; this and other possible reasons for failure are discussed. PMID- 14543706 TI - The isolation and characterization of antibiotic biosynthesis genes. AB - Antibiotic biosynthesis pathways are found in a broad range of Gram positive prokaryotes, a smaller range of Gram negative prokaryotes and a limited range of eukaryotes. A variety of techniques can be used to identify the genes involved in the biosynthesis of these compounds ranging from genetic complementation and interspecific gene transfer to polymerase chain reaction amplification and transposon mutagenesis. The dissection of these cloned pathways and the understanding of their structure and regulation has led to insights into the structure and function of antibiotic biosynthesis genes. With new knowledge of the structural similarities and relationships between related antibiotic biosynthesis pathways, the possibility of directed manipulation of specific genes to allow synthesis of novel antibiotics is now possible. PMID- 14543707 TI - Biotransformations of rifamycins: process possibilities. AB - Rifampicin, an important antibiotic, is manufactured by chemical conversion of rifamycin S which is obtained by the chemical modification of rifamycin B. Rifamycin B is a product of Nocardia mediterranei fermentations. The chemical conversion of rifamycin B to rifamycin S has many disadvantages: Strong acidic conditions are required, heavy foam formation accompanies transformation and the yields are low. This review highlights the developments in alternative, biochemical transformations using enzymes and cells; the main focus is on transformations carried out by rifamycin oxidase. PMID- 14543716 TI - Microbial cultures in the utilization of cellulosic materials. AB - This review elaborates on the most recent microbial development in saccharification of cellulose and cellulase formation. A particular highlight is a new genetic-immunochemical approach investigating the mechanism of adhesion of bacterial cellulase to cellulose during cellulose conversion. New developments and recent reviews in hemicellulose and lignin degradation are also covered. PMID- 14543717 TI - Effective diffusivity of oxygen in microbial pellets. AB - In a typical submerged aerobic fermentation with microbial pellets, the effective diffusivity of oxygen in the pellets is probably the most important, yet most difficult transport property to characterize experimentally. Its values directly indicate the efficiency or deficiency of oxygen to individual cells, and thus the biological activity of the microorganisms. In the past, it was not possible to assess reliably the effective diffusivity of oxygen in pellets due to several reasons. Firstly, most oxygen electrodes available were coarse, and hence not suitable for in situ measurements. Secondly, there was a lack of methods rigorous enough to characterize the structure of the microbial pellets. A state-of-the-art review of the literature relating to the feature subject is presented. Emphasis is laid upon development and evolution of the means for quantitative characterization of the effective diffusivity of oxygen in microbial pellets. PMID- 14543718 TI - Continuous cultivation of microorganisms. AB - This review presents selected studies on continuous cultivation including the genetics and selection of microorganisms, production of biomass, liquid fuels and other products, published in 1982-1984 and bearing a relationship to biotechnology. PMID- 14543719 TI - Pretreatment for enhanced hydrolysis of cellulosic biomass. AB - This review will cover a number of physical and chemical pretreatment methods for cellulosic substrates which enhance their hydrolysis by cellulase or consumption by microorganisms. While the emphasis is on the literature of the last two years, some earlier work is cited which has influenced the work in the pretreatment field. In order to interpret the effects of a pretreatment method, emphasis in the past has been on crystallinity index (CI) and lignin content. Although these parameters happen often to correlate with the rate or extent of hydrolysis, it is suggested that a more basic parameter is the pore size distribution of the wet substrate and the associated surface area available to the cellulase that is the major factor in determining the effectiveness of a pretreatment method. PMID- 14543720 TI - Magnetic separation in biotechnology. AB - New developments in magnetic labelling techniques for cells and microspheres have extended the useful range of magnetic separation, particularly high gradient magnetic separation, into biotechnical areas. The basic magnetic principles involved are reviewed and representative samples of labelling techniques and results drawn from the past three years are presented. Illustrative examples of large scale operation in other industries are also presented, demonstrating the potential of the biotechnological applications. PMID- 14543721 TI - Anaerobic waste stabilization. AB - The present knowledge of the microbiology, physiology and regulation of anaerobic digestion in conventional or advanced processes is reviewed. In all systems the carbon flow from biopolymers to biogas is determined by syntrophic interactions of fermentative or acetogenic bacteria with methanogens at the level of interspecies hydrogen transfer. Inhibitors or heavy metal ions may interfere at different levels. The stabilization of waste at mesophilic and thermophilic temperatures is compared and the process stability as well as the inactivation of pathogens is discussed. Characteristics of conventional digestion systems and of recently developed advanced processes with solids and liquids uncoupling are compared and selection criteria with respect to the type of sludge are outlined. Areas of future research for a better understanding of the biochemistry, the physiology and the regulation of the degradation of pollutants are suggested. PMID- 14543722 TI - Mycorrhizae and their potential use in the agricultural and forestry industries. AB - Mycorrhizal fungi associated with plant roots increase the absorption of nutrients, particularly phosphorus, and thus enhance the growth of crop plants and trees. Vesicular-arbuscular mycorrhizae (VAM) occur in approximately 90% of all vascular plants including most of the important agricultural species, whereas ectomycorrhizae are found in most of the economically important tree species of the temperate regions of the world, and in some tropical trees. These symbiotic associations are, therefore, important in crop and biomass production. For this reason they are receiving considerable attention in agriculture and forestry. Currently, VAM are utilized in fumigated soils, greenhouse crops, and in the reclamation of disturbed sites. Ectomycorrhizae are employed in the establishment of trees in nurseries, in reforestation programs, and in the production of containerized seedlings. Production of VAM and ectomycorrhiza inoculum for large scale projects is now feasible but many basic questions related to persistence of these fungi in field situations, competition with other microorganisms, and particularly the most efficient fungi to use for particular hosts remain largely unanswered. PMID- 14543736 TI - Physical methods for the transformation of plant cells. AB - Transfer and expression of foreign genes in adult plants and their progeny has been achieved by acceleration of DNA-coated particles or microinjection techniques. Cultured cells or embryoids served as targets for the introduction of marker genes that were stably expressed in the nucleus or the chloroplast. Cloned genes from the maize anthocyanin pathway were regulated in a tissue-specific manner when transferred into maize by particle acceleration. In spite of these successes, stable transformation efficiency was low due to uneven particle distribution and cell death after bombardment. Transferred genes did not always segregate in a Mendelian fashion in the succeeding generation, and additional efforts of embryo rescue or shoot grafts were needed to obtain viable progeny from original transformants. New technical advances such as the helium-driven particle gun may improve transformation rates in the future, but some problems of cell manipulation remain. PMID- 14543737 TI - Cultivation techniques for the production of ectomycorrhizal fungi. AB - Ectomycorrhizal fungi have a valuable role to play in current agricultural practices. In order to produce inocula for use in laboratory, nursery and field trials it is first necessary to isolate the organism, grow it in axenic culture and produce sufficient quantities of the fungus. The methods currently employed to cultivate this group of fungi are considered. PMID- 14543738 TI - Biotechnology and aquaculture: the role of cell cultures. AB - Cell culturing complements recombinant DNA technology in the application of biotechnology to aquaculture. Cell cultures can be prepared from the three main groups of multicellular organisms in aquaculture: fish, shellfish, and seaweeds. These cultures can contribute indirectly to the successful farming of these organisms by providing basic insights into how their growth, reproduction, and health can be understood and manipulated. Finally, they can be a direct source of diverse biochemical products for use in aquaculture, medicine and the food industry. PMID- 14543739 TI - Production scale insect cell culture. AB - Insect cells in culture are currently commanding great interest as superior hosts for the efficient production of biologicals with applications in health care and in agriculture. Insect cell culture is ripe for scale-up technologies, in order to meet future projected production requirements of (a) insect viruses used as bioinsecticides and (b) recombinant proteins of therapeutic potential for humans and animals. The single most prominent system used in research-based and in commercial insect cell culture today involves lepidopteran cells transfected with baculovirus expression vectors for abundant formation of recombinant biologicals. However, dipteran insect cell lines also are beginning to emerge as useful tools in biotechnology. Current practices in bioprocess development using insect cell culture, advances in media formulation and in insect cell bioreactor design, and emerging trends are presented and critically evaluated. PMID- 14544047 TI - Ecology and genetics of tropical rhizobia species. AB - Biological nitrogen fixation (BNF) technology with special reference to Rhizobium legume symbiosis is growing very rapidly with the hope of combatting world hunger by producing cheaper protein for animal and human consumption in the Third World. One can see rapid progress made in the biochemistry and molecular biology of symbiotic nitrogen fixation in general; however, less progress has been made on the ecological aspects despite the fact that an enormous amount of literature is available on inoculation problems and on agronomic aspects of symbiotic nitrogen fixation. So far most information on Rhizobium concerns fast-growing rhizobia and their host legume. Although it is essential that food production using BNF technology should be maximized in the Third World, the least work has been done on slow-growing rhizobia, which are generally found in tropical and sub-tropical soils. The majority of the developing countries are in tropical and sub-tropical regions. Except for R. japonicum, a microsymbiont partner of soybean (Glycine max), the majority of the slow-growing rhizobia belong to the cowpea group, and we refer to cowpea rhizobia as tropical rhizobia species. In this review we have tried to consolidate the recent progress made on ecology and genetics of tropical rhizobia. By using recombinant DNA technology techniques it is expected that super strains of rhizobia with desirable characteristics can be produced. One must evaluate the efficiency and effectiveness of these genetically manipulated laboratory strains under field conditions. In conclusion, if one aims at combatting hunger in the Third World using BNF technology, an intensive research programme on fundamental and applied aspects of tropical rhizobia species is suggested. This involves close cooperation between molecular biologists and microbial ecologists. PMID- 14544048 TI - Antibiotic biosynthesis. PMID- 14544049 TI - Biotechnological uses of cyanobacteria. AB - Cyanobacteria (blue-green algae) are O(2)-evolving photosynthesizing prokaryotes that have an extensive history of use as a human food source and as a fertilizer in rice fields. They have also been recognized as an excellent source of vitamins and proteins and as such are found in health food stores in North America and elsewhere. Cyanobacteria have a great deal of potential as a source of fine chemicals, as a biofertilizer and as a source of renewable fuel. This potential is being realized as data from research in the areas of the physiology and chemistry of these organisms are gathered and as the knowledge of cyanobacterial genetics and genetic engineering increases. We review, here, the present (and possible future) uses of cyanobacteria and assess the state of the art with respect to the genetic manipulation of cyanobacteria. PMID- 14544050 TI - Biosensors: recent trends. AB - One of the major bottlenecks in automation and process control of industrial bioprocesses is the lack of suitable sensing devices to accurately measure the concentrations of biomolecules. The measurement of ions (e.g., H(+), NH(4)(+)) and gases (e.g., O(2), CO(2), NH(3)) using standard ion-selective and gas sensing electrodes respectively, is well established. Chemical analysis of biomolecules off-line is generally unreliable, labour intensive and may lead to contamination of the biological systems. Problems of maintaining sterile conditions are especially important when dealing with slow growing mammalian or plant cells in culture. Active research in the development of biosensors for monitoring fermentation processes, food production and pollution control, and for medical and veterinary applications is currently underway. This paper reviews recent approaches toward the development of biosensors which involve a biochemical interaction to measure the concentrations of biomolecules, primarily for the on line monitoring and control of fermentation processes. PMID- 14544051 TI - Transport phenomena, reactor design and scale-up. AB - Mixing in biological reactors is used to improve mass transfer and provide proper micro-scale and macro-scale shear rates for effective process results. Reactors may be mixed by impellers on rotating shafts, or may be of the flow contactor type such as packed columns, bubble columns or airlift circulators. In addition, review of kinetics can tell the process performance based on various kinds of mixing conditions. Several interesting and unique mixing studies are also included where appropriate. PMID- 14544242 TI - Development of DNA-mediated transformation systems for plants. AB - The genetic engineering of plants by DNA-mediated gene transfer requires that efficient transformation systems be developed. Considerable progress has been made in manipulating the Ti plasmid of Agrobacterium tumefaciens as a vehicle for delivery of foreign genes into protoplasts of dicotyle-donous plants. Part of the Ti plasmid, the T-DNA, can be incorporated into the genome of the host cell; the T-DNA can carry a foreign DNA sequence which co-integrates with it; under normal conditions, the tumorigenic-causing portion of the T-DNA can be inactivated so that transformed protoplasts can be regenerated and T-DNA with an inserted foreign gene can be stably maintained during regeneration, meiosis and gamete formation. A foreign gene has yet to be expressed in regenerated plants although a T-DNA gene for opine synthesis can function in regenerates. Developing a more ubiquitous transformation system for monocotyledons is further from fruition. Based on transformation systems for simple eukaryotic organisms, it is reasonable to expect that a DNA vector which is capable of amplifying a novel plant gene and which contains both a drug resistance marker to facilitate the selection of transformed plant protoplasts and a species-specific autonomously replicating sequence to ensure the stable maintenance of the input gene in the recipient cell can be constructed. PMID- 14544243 TI - Transport phenomena, reactor design and scale-up. AB - This review will cover the area of impeller-mixed stirred-tank reactors. In addition, it will consider bubble columns, in which air or gas is passed up a liquid filled column through distribution plates covering the full area of the column, and also airlift reactors, in which the air is confined in a channel by means of a loop or draft tube designed to impart a certain type of overall circulatory pattern to the entire tank. There is considerable interest in the kinetics inside the solid part of various kinds of immobilized solid pellet type of enzymes and catalysts. The use of these particles in fixed bed reactors is also covered. PMID- 14544244 TI - Fermented foods, feeds and beverages. AB - There has been a proliferation of books and papers dealing with the indigenous fermented foods/beverages of the world. It is anticipated that these foods/beverages will play an ever-increasingly important role in feeding both the developing and the developed world as population increases from approximately 4.5 billion to 6 billion by the year 2000 and to 8 to 12 billion people in the 21st century. The indigenous fermented foods consist of microbial protein grown on edible substrates. Microbial or single cell protein (SCP) per se continues to receive research and development attention. It is likely to play an important role in feeding animals in the future when it becomes competitive with soy protein. It may play a direct role in feeding humans in the future after its safety for feeding animals has been adequately demonstrated and it has been shown that it can be processed into foods acceptable to humans. At the present time, mushrooms, a form of microbial protein highly acceptable to humans, which can be grown readily on ligno-cellulosic and other agricultural and food processing wastes, offer considerable opportunity for expanding man's food supply. PMID- 14544245 TI - Fuels and industrial chemicals. AB - The favorite subject of recent literature on biotechnical processes has been ethanol fermentation. This review covers a number of new technics developed, including immobilized biocatalyst technology and bacterial fermentations. Reference is also made to recent work on whey, starch, inulin, and cellulosic materials as substrates for ethanol production. Renewed interest in acetonebutanol fermentation for solvent and liquid fuel production has also been clearly evident during the last two years. Biotechnical production of organic acids has been considered as an alternative route to chemical feedstocks. New developments in amino acid, methane, hydrogen, and hydrocarbon production, and on hydrocarbon oxidation are also briefly covered. PMID- 14544246 TI - Biotechnology developments for the treatment of toxic and inhibitory wastewaters. AB - The cost-effectiveness of biological processes has encouraged many researchers to consider biotreatment for the stabilization of toxic or recalcitrant wastewaters. However, to ensure adequate removal of trace contaminants and satisfactory performance with high strength inhibitory industrial wastewaters, conventional biotechnology is being re-evaluated. This review summarizes selected recent contributions to the development of appropriate biotechnology for toxic wastewater treatment. Microbiological constraints and potential solutions are examined. Assessments of conventional biological processes for contaminant control are reviewed, and several new developments in bioreactor design for inhibitory wastes are presented. PMID- 14544247 TI - Biotechnology applied to mining of metals. AB - The present review describes the advances achieved during the last two years in the application of biotechnological principles in the extraction of metals from ores and minerals. Despite the fact that this branch of science is very young and many details are yet to be understood, the microbes are applied at commercial levels especially for the extraction of copper and uranium from low-grade ores. The technique is far from being developed to its full potential and it is generally recognized to be a technology of the future. The studies involved are complex and multidisciplinary in nature. PMID- 14544248 TI - Diagnostic and biomedical applications: monoclonal antibodies. PMID- 14544531 TI - Inaugural editorial. PMID- 14544532 TI - Evaluation of methods for detecting ecological effects from genetically engineered microorganisms and microbial pest control agents in terrestrial systems. AB - This report summarizes and evaluates research from several laboratories that deals with the detection of ecological effects induced through exposure of microbes or plants to genetically engineered microorganisms (GEMs) and microbial pest control agents (MPCAs). Some 27 potential endpoints for measuring effects have been studied. Perturbations induced by GEMs have been detected in about one half of these endpoints. Detectable effects have been recorded for over half of the 16 species of bacteria and fungi studied. The effects caused by GEMs and MPCAs include inhibition of beneficial mycorrhizal fungi growing on Douglas fir seedling roots, depression in plant root and shoot growth, inhibition of predatory soil protozoa, accumulation of a toxic metabolite during biodegradation that inhibits soil fungi, increased microbial community respiration due to rapid lignin breakdown in soil, and the displacement of a broad group of gram-negative bacteria that inhabit the root surface of cereal crops. These effects were usually, but not always, of short duration. However, some of the changes were irreversible during the observation time of days, weeks, or in one case, months. PMID- 14544533 TI - High-level expression of foreign genes in Hansenula polymorpha. AB - The methylotrophic yeast Hansenula polymorpha belongs to a limited number of non Saccharomyces yeast species used as hosts for heterologous gene expression. It has successfully been applied for the production of hormones, antigens and enzymes. The system excells by mitotically stable recombinant strains, high productivity and faithful processing of the produced polypeptides. The favourable characteristics of this microorganism for protein production at an industrial scale are described in the following article focusing on some recent representative examples. PMID- 14544534 TI - Production of Phanerochaete chrysosporium lignin peroxidase. AB - Liginin peroxidase (ligninase) of the white rot fungus Phanerochaete chrysosporium Burdsall was discovered in 1982 as a secondary metabolite. Today multiple isoenzymes are known, which are often collectively called as lignin peroxidase. Lignin peroxidase has been characterized as a veratryl alcohol oxidizing enzyme, but it is a relatively unspecific enzyme catalyzing a variety of reactions with hydrogen peroxide as the electron acceptor. P. chrysosporium ligninases are heme glycoproteins. At least a number of isoenzymes are also phosphorylated. Two of the major isoenzymes have been crystallized. Until recently lignin peroxidase could only be produced in low yields in very small scale stationary cultures owing to shear sensitivity. Most strains produce the enzyme only after grown under nitrogen or carbon limitation, although strains producing lignin peroxidase under nutrient sufficiency have also been isolated. Activities over 2000 U dm(-3) (as determined at 30 degrees to 37 degrees C) have been reported in small scale Erlenmeyer cultures with the strain INA-12 grown on glycerol in the presence of soybean phospholipids under nitrogen sufficiency. In about 8 dm(3) liquid volume pilot scale higher than 100 U dm(-3) (as determined at 23 degrees C) have been obtained under agitation with immobilized P. chrysosporium strains ATCC 24725 or TKK 20512. Good results have been obtained for example with nylon web, polyurethane foam, sintered glass or silicon tubing as the carrier. The immobilized biocatalyst systems have also made large scale repeated batch and semicontinuous production possible. With nylon web as the carrier, lignin peroxidase production has recently been scaled up to 800 dm(3) liquid volume semicontinuous industrial production process. PMID- 14544535 TI - Enzyme stability in downstream processing. Part 1: enzyme inactivation, stability and stabilization. AB - In biotechnological recovery processes the instability of the product can lead to large losses in the sequence of recovery processes needed to purify the product. As the cost of the final active product is strongly dependent on the recovery yield, this will lead to an increase in product cost. Therefore knowledge of factors that influence stability is important. This Part 1 contains a review on the factors that influence stability. As stability is very important in enzyme purification this review deals about enzymes and their ability to retain catalytic activity. Inactivation mechanisms and agents are discussed. A short review is given of enzyme structure and stability. This is followed by stabilization strategies and methods. PMID- 14544536 TI - Enzyme stability in downstream processing. Part 2: quantification of inactivation. AB - In biotechnological recovery processes the instability of the product can lead to large losses in the sequence of recovery processes needed to purify the product. As the cost of the final active product is strongly dependent on the recovery yield, this will lead to an increase in product cost. Therefore knowledge of factors that influence stability is important. This Part 2 provides the basic principles for design and operation of processes in which inactivation takes place. Simple kinetics and reactor modelling are discussed. These are applied to a number of unit operations: cell disruption, membrane filtration, drying and reversed micellar extraction. It is thus shown that the basic tools for modeling of biochemical processes provide us with the data needed for optimal process design and operation. PMID- 14544806 TI - Development of recombinant vaccines against bluetongue. AB - Bluetongue virus is the aetiological agent of bluetongue, a disease of domestic and wild ruminants. Twenty-four serotypes are recognized. Novel subunit vaccines, that complement existing modified live polyvalent vaccines, are being developed. Serotype-specific viral neutralizing antibodies that are able to protect sheep against virulent homologous virus challenge can be induced by immunizing with the BTV outer capsid protein VP2 purified from virions or with VP2 expressed by baculovirus recombinants. Presentation of VP2 on virus-like particles, which assemble upon co-expression of the four major structural viral proteins (VP2, VP5, VP3 and VP7), improves the protective effect of VP2. Sheep immunized with core-like particles, comprised of VP3 and VP7, developed only limited clinical signs after virulent virus challenge, demonstrating that not only the outer capsid proteins, but also the core proteins are involved in protection against bluetongue. PMID- 14544807 TI - Reverse transcription-polymerase chain reaction: an overview of the technique and its applications. AB - The polymerase chain reaction (PCR) has galvanized molecular biologists by virtue of its ability to provide them with large quantities of any desired fragment (up to 11kb) of DNA. This power combined with its flexibility has also inspired many useful applications, including new methods of DNA sequencing, cloning and mutagenesis. One logical variation of PCR has been its application to the detection and analysis of messenger RNA by the addition of a reverse transcription step prior to performing PCR. Due to the exquisite sensitivity of PCR, reverse transcription-PCR (RT-PCR) has been used to characterize mRNAs previously undetectable by established methods of RNA analysis such as Northern hybridization and RNase protection assays. Furthermore, its capacity as a method of quantitative analysis is currently being developed. RT-PCR has also been used to diagnose the presence of certain diseases. Recently, RT-PCR has been employed to identify and isolate genes that are differentially expressed in different cells or environmental conditions. PMID- 14544808 TI - Improvement of downstream processing of recombinant proteins by means of genetic engineering methods. AB - The rapid advancement of genetic engineering has allowed to produce an impressive number of proteins on a scale which would not have been achieved by classical biotechnology. At the beginning of this development research was focussed on elucidating the mechanisms of protein overexpression. The appearance of inclusion bodies may illustrate the success. In the meantime, genetic engineering is not only expected to achieve overexpression, but to improve the whole process of protein production. For downstream processing of recombinant proteins, the synthesis of fusion proteins is of primary importance. Fusion with certain proteins or peptides may protect the target protein from proteolytic degradation and may alter its solubility. Intracellular proteins may be translocated by means of fusions with signal peptides. Affinity tags as fusion complements may render protein separation and purification highly selective. These methods as well as similar ones for improving the downstream processing of proteins will be discussed on the basis of recent literature. PMID- 14544809 TI - Factor VIII: structure, function and analysis. AB - Factor VIII is an important blood coagulation protein whose genetic deficiency leads to the serious bleeding disorder, classic haemophilia (haemophilia A). Here we review the structure, function and analysis of this protein for diagnostic and therapeutic applications. Because factor VIII is tightly associated with von Willebrand factor some recent work on the latter is also considered so as to clarify the relationship between them. PMID- 14544810 TI - Development of thrombolytic agents. AB - Despite their widespread use in patients with acute myocardial infarction, all currently available thrombolytic agents suffer from a number of significant limitations, including resistance to reperfusion, the occurrence of acute coronary reocclusion and bleeding complications. Furthermore, the therapeutic use of plasminogen activators as thrombolytic agents requires intravenous infusion of relatively large amounts of material. Therefore, the quest for thrombolytic agents with a higher thrombolytic potency, specific thrombolytic activity and/or a better fibrin-selectivity continues. Several lines of research towards improvement of thrombolytic agents are being explored, including the construction of mutants and variants of plasminogen activators, chimeric plasminogen activators, conjugates of plasminogen activators with monoclonal antibodies, or plasminogen activators from animal or bacterial origin. PMID- 14545005 TI - Biological bleaching of chemical pulps: a review. AB - The pulp and paper industry is implementing changes in the bleaching process to minimize the use of chlorine in order to satisfy regulatory and market demands. Biotechnology has a potentially important role to play in providing alternatives to conventional chlorine bleaching of chemical pulps. The current developments in fungal, enzymatic and biomimetic bleaching are reviewed here within an engineering context. PMID- 14545006 TI - Protoplast fusion: a tool for intergeneric gene transfer in bacteria. AB - Protoplasts can be isolated from bacterial cells by digestion of the cell wall with the help of lysozyme in presence of osmotic stabilizers. Fusion of protoplasts can be induced by chemical fusogens like polyethylene glycol. The electrofusion technique has been reported in bacteria in which the fusion frequency is much higher than that obtained by PEG induced protoplast fusion. This technology allows recombination to take place not only between related species but also between unrelated genera and is of great potential in the breeding and improvement of industrial strains. This review includes the information and developments on the protoplast fusion in bacteria with special reference to genetic recombination by protoplast fusion between phylogenetically unrelated bacteria. PMID- 14545007 TI - Biologically active secondary metabolites from myxobacteria. AB - New chemical structures with proven biological activity still are badly needed for a host of applications and are intensively screened for. Suitable compounds may be used as such, or in the form of their derivatives or, equally important, may serve as lead compounds for designing synthetic analogs. One way to new compounds is the exploitation of new producer organisms. During the past 15 years the myxobacteria have been shown in our laboratories to be a rich source of novel secondary metabolites, many of the compounds showing interesting and sometimes unique mechanisms of action. About 50 basic structures and nearly 300 structural variants have been elucidated, and almost all of them turned out to be new compounds. Several myxobacterial substances may have a good chance of an application. PMID- 14545008 TI - Animal cells in turbulent fluids: details of the physical stimulus and the biological response. AB - Animal cells in large scale bioreactors are subjected to a variety of fluid forces for which they are not adapted by evolution. In severe cases the result is cell death, but under more modest agitation conditions an increasing number of nonlethal responses affecting growth rate, metabolism, and product formation have been reported. The forces causing these responses have not been characterized because particle-turbulence interactions are extremely complex. The current understanding of the microscopic structure of turbulence in an infinite liquid and in boundary layers shows that an average shear stress alone is not likely to be adequate to describe the bioreactor environment. Combining knowledge of the physical stimuli and the biological responses will lead to better ways of limiting cell damage and possibly to using physical stresses as a means of specifically modifying cell behavior. PMID- 14545371 TI - Use of synthetic DNA in expression of foreign proteins. AB - Synthetic DNAs and oligonucleotides, which can be prepared conveniently by combining chemical synthesis and enzymatic methods, have been used extensively in recombinant DNA research. Examples include total gene synthesis, probes for the isolation of specific genes from cDNA or genomic libraries, linkers containing specific restriction sites for cloning, primers for DNA and RNA sequencing, and primers for the construction of specific mutations (either deletion, insertion or point mutations) by oligonucleotide-directed site-specific mutagenesis. This article reviews recent advances in the chemical and enzymatic synthesis of oligo- and polynucleotides and the application of synthetic DNA to the expression of foreign proteins. The synthesis of genes, including structural genes and regulatory genes are reviewed. Oligonucleotide-directed site-specific mutagenesis and use of synthetic DNA to optimize foreign protein expression are also discussed. PMID- 14545372 TI - Biotechnology for phosphorus removal during wastewater treatment. AB - Advanced biological wastewater treatment for the removal of phosphorus in excess of the normal metabolic requirements of activated sludge type processes has been developed as an alternative to chemical addition. Current laboratory and pilot plant investigations have confirmed that a preliminary anaerobic zone and plug flow type configuration are necessary for good enhanced biological phosphorus removal. Nitrate in the anaerobic stage inhibits the process whereas acetate enhances phosphorus uptake. The bacteria probably responsible are of the Acinetobacter genus and the presence of stored polyphosphate within these bacteria has been demonstrated. It has also been shown that pure cultures of Acinetobacter do not necessarily take up soluble substrate as phosphate is released during the anaerobic phase, in contrast to the current proposed mechanism, and that in certain cases natural chemical precipitation could make a significant contribution towards overall phosphorus removal. Several studies of pilot and full-scale plants have been reported. PMID- 14545373 TI - Influence of chemical modification on enzyme inactivation kinetics and stability. AB - Enzymes are placed in different categories depending on the effect of chemical modification on their inactivation kinetics and residual activity. This is done using a series-type mechanism involving degraded but stable enzyme states. The major distinction in the three basic categories is the effect of modification on residual activity. Each category is further sub-divided depending on the effect of modification on the values of the deactivation rate constants. The classification provides for a framework for comparison of a wide variety of enzyme deactivation data. Structure-function relations are provided wherever possible. PMID- 14545374 TI - Expression of recombinant DNA functional products in Escherichia coli anucleate minicells. AB - This review covers the use of anucleate minicells of Escherichia coli for expressing the recombinant DNA encoded proteins. We briefly discuss the methods being used for preparation of anucleate minicells, incorporation of cloned DNA and assessment of gene expression. While the largest use has been that of microbially derived cloned functional DNA, examples of eukaryotic gene product synthesis have also been reviewed. This technology may represent some interesting commercial opportunities. PMID- 14545375 TI - Zymomonas mobilis: a bacterium for ethanol production. PMID- 14545376 TI - Variation from plant tissue cultures: biotechnological application to improving salinity tolerance. AB - Breeding for salt tolerance in crop plants is envisaged as one way to combat a worldwide problem of increasing soil salinity in agricultural land. Tissue culture techniques may prove valuable as a means of achieving this goal. In this review, reports of the selection and characterization of plant cell cultures tolerant to excess salt are assessed, in the context of variability from tissue culture and the significance of cellular physiological adaptation to salinity. The examples of plant regeneration from salt grown cell cultures are also outlined, with emphasis on correlation to the effect of salt on cell cultures, genetic variability for salt tolerance in vitro, and the value of regenerates in the development of salt tolerant plants. PMID- 14545663 TI - Solid state fermentation: acid protease production in controlled CO2 and O2 environments. AB - The effect of the partial pressure of O(2) and CO(2) on the acid protease production in solid state fermentation by Aspergillus niger on wheat bran was studied. A fermentation system was used, which allowed on-line reactor measurements and continuous data acquisition of pH, temperature, gas flow, pressure drop and CO(2) production. Six paired combinations of CO(2) and O(2) concentrations were studied. The results showed a direct relationship between pressure drop, production of CO(2) and temperature increase. The pH evolution patterns were similar in all cases but different if the measurements were made on line or on a liquid homogenate of the fermented substrate. Acid protease production was increased when the gas had 4% CO(2), (vol/vol), and it reached its highest level, a 43% increase over air, with a mixture of 4% CO(2) and 21% O(2). The protease production was strongly related to the mold metabolic activity as represented by the total CO(2) evolved. PMID- 14545664 TI - Downstream processing of microbial rennet from solid state fermented moldy bran. AB - In recent years due to acute shortage of calf-rennet, microbial rennets seem to be an effective alternative and are commercially produced. Mucor miehei was cultivated under the solid state fermentation conditions, and the moldy bran was extracted using a semicontinuous multiple contact forced percolation method. The treated extract was then filtered through 5% R16 clay which enabled easy and efficient removal of impurities such as lipase and protease without involving costly chemical treatments. The ethyl alcohol precipitated enzyme was dried and made into powder form having activity of 1.5 x 10(5)Soxhlet units/gm. PMID- 14545665 TI - Effect of culture conditions on lipolytic enzyme production by Penicillium candidum in a solid state fermentation. AB - Lipolytic enzymes were produced using wheat bran as substrate in a solid state fermentation with Penicillium candidum. The best production of lipolytic activity occurred at 29 degrees C. One hundred micromoles of free butyric acid (FBA) was released from tributyrin by 1 mL of cell free supernatant in the absence of control of environmental relative humidity. When a closed chamber saturated with water vapour was used the lipolytic activity increased to 320 micromoles of free butyric acid. The best initial reaction pH was 7.0. The highest activity, 480 micromoles of FBA, was obtained at a moisture content of 67.5 % of saturation. PMID- 14545666 TI - Production of novel oligosaccharide oxidase by wheat bran solid-state fermentation. AB - A search for oxidases that catalyze the oxidation of oligosaccharides has resulted in the isolation of several soil-derived fungus strains which produced novel oligosaccharide oxidases with different substrate specificity on wheat bran solid culture. One of these oxidases produced by Acremonium strictum T1 strain has been characterized. This enzyme showed high reactivity toward maltose, lactose, cellobiose and maltooligosaccharides composed of up to seven glucose units, and was named as glucooligosaccharide oxidase based on its substrate specificity. Strain T1 was subjected to a strain improvement program, and an enzyme hyper-producing mutant strain T1-38 was selected. This mutant strain produced glucooligosaccharide oxidase 75 times higher than the wild type strain T1. When cultivated in a solid medium comprised of 1 part of wheat bran and 1 part of water (w/w), enzyme activity reached a maximum level of 6 units per g of culture medium after 4 days cultivation. Characteristics of the enzyme including the substrate specificity were compared with two other novel oligosaccharide oxidases isolated in this laboratory. Batch type conversion of lactose to lactobionic acid using crude enzyme was also discussed. PMID- 14545667 TI - New approach for selecting pectinase producing mutants of Aspergillus niger well adapted to solid state fermentation. AB - The aim of this paper is to review and study a new approach for improving strains of Aspergillus niger specially adapted to produce pectinases by Solid State Fermentation (SSF) with materials having low levels of water activity (a(w)), i.e., coffee pulp. Special emphasis is placed on the use of two antimetabolic compounds: 2-deoxy-glucose (DG) and 2,4-dinitro-phenol (DNP) combined with a water depressant (ethylene glycol = EG) in order to put strong selection pressures on UV treated spores from parental strain C28B25 isolated from a coffee plantation. Such a strain was found to be DG sensitive. Results suggested the existence of a reciprocal relation between adaptation of isolated strains to SSF or to Submerged Fermentation (SmF) systems. Preliminary physiological analysis of isolated strains showed that at least some few initially DG resistant mutants could revert to DG sensitive phenotype but conserving increased pectinase production. Also it was found that phenotype for DNP resistance could be associated to changes of DG resistance. Finally, it was found that low levels of a(w) produced by adding 15% EG to agar plates, were a significant selection factor for strains well adapted to SSF system. PMID- 14545668 TI - Properties of thermostable hemicellulolytic enzymes from Thermomonospora strain 29 grown in solid state fermentation on coffee processing solid waste. AB - During decaffeination of Coffee Processing Plant Solid Wastes (CPSW) by actinomycetes, Thermomonospora, Strain 29 exhibited high titers of cellulase and xylanase. This organism, originally isolated on soybean seed coat was grown in solid state fermentation on CPSW supplemented with mineral salts. Enzymes recovered were arabinosidase, xylanase, and beta-D-xylosidase. Higher activity of the former two enzymes was in the extracellular broth, whereas the beta-D xylosidase activity was highest in the cell fraction. The enzymes were characterized after precipitation with (NH(4))(2)SO(4), dialysis, and gel filtration. Production of all three enzymes was inhibited by monomeric sugars and sugar alcohols but not by arabinoxylan, xylans, or xylan containing water insoluble carbohydrates. The optimum pH for the activity was 6.5, 7.0, and 7.5 for beta-xylosidase, xylanase and arabinosidase (alpha-L-arabinofuranosidase, alpha-arabinosidase, alpha-L-arabinosidase) respectively. These enzymes were stable in the pH range of 6.5 to 8.0. All three enzymes were thermostable up to 80 degrees C. At 55 degrees C, arabinosidase had the longest half life of 120 h. However, at 40 degrees C, xylanase had the longest half life (504 h). At either temperature, beta-D-xylosidase had the shortest half life. The molecular weights (kDa), and Kms (mM) were estimated to be 95, 0.27; 45, 12.4; and 106, 0.67 for arbinosidase, xylanase, and beta-xylosidase respectively. Step wise addition of the three enzymes showed higher saccharification of lignocellulosics. PMID- 14545669 TI - Fermentation of cellulosic materials to mycoprotein foods. AB - A new bioprocess is described in which a cellulolytic, food-grade fungus Neurospora sitophila converts cellulosic materials to protein-rich products for food and fodder. The optimal conditions for the conversion are identified: 35-37 degrees C temperature, pH 5.5, 2.35 ms(-1) agitator tip speed. Scale-up of the production process to 1,300 L is reported. The mycoprotein production data on several types of cellulosic materials (sugarcane bagasse, corn stover, wood cellulose) are presented. The performance of N. sitophila is found to compare favourably with that of Chaetomium cellulolyticum, another cellulolytic organism previously reported on by us. PMID- 14545670 TI - Tempe fermentation: some aspects of formation of gamma-linolenic acid, proteases and vitamins. AB - During a tempe fermentation the concentrations of linoleic, and alpha-linolenic acids (ALA) decreased while the concentration of oleic acid increased. During fatty acid synthesis Rhizopus sp. produced only gamma-linolenic acid (GLA) instead of ALA. The amount of GLA in tempe were influenced by varying external parameters. The proteolytic capacity of 36 strains of the genus Rhizopus isolated from Indonesian tempe or tempe inocula was examined. There was a distinct increase in the amount of free amino acids during tempe fermentation. Fermentations with mixed populations of bacteria and Rhizopus yielded a lower level of free amino acids, but an increase in total amount of amino acids. In comparison to intracellular, and extracellular proteases the proteases of the cell wall fraction are most responsible for proteolytic capacity of the different Rhizopus strains. Two isolated strains of Citrobacter freundii were found to be the best vitamin B(12) producers during the soaking of soybeans. In the solid substrate fermentation the Rhizopus molds formed vitamin B(6), riboflavin, and nicotinic acid. The addition of bacteria to the solid substrate fermentation resulted in a strong increase of active vitamin B(12) in tempe. In the presence of the Rhizopus mold, the vitamin B(12) formation by C. freundii was three times higher than that of a fermentation without the mold. PMID- 14545671 TI - Protein enrichment of sweet potato residue with co-culture of amylolytic fungi by solid-state fermentation. AB - Protein enrichment of sweet potato residue with amylolytic moulds by solid-state fermentation was higher than that obtained with amylolytic yeasts. The optimum initial moisture content for protein enrichment was 66% to 75%. Incrementally added nitrogen sources to the culture at zero time and at 24 h considerably improved the final protein content. During the cultivation, the moisture, ash and ATP contents increased, while the pH value decreased. A 1:1 co-culture of amylolytic mycelial fungi yielded a product with 32.4% crude protein after 4 days incubation at 30 degrees C. PMID- 14545672 TI - Bioreduction of prochiral ketones with yeast cells cultivated in a vibrating air solid fluidized bed fermentor. AB - A brief review of fluidized bed fermentors and of bioreduction of prochiral ketones by yeast cells is presented. Cultivation of yeast cells, Saccharomyces cerevisiae HUT 7099, in a vibrating fluidized bed and the bioreduction of ethyl acetoacetate by the cells are described. The cultivation of the cells in the fermentor was successfully performed at relatively low moisture content, about 40 % on wet basis. The cell size decreased and the shape changed from ellipsoid to spherical after the logarithmic growth phase. The biocatalytic performance of yeast cells cultivated in submerged, static solid, and fluidized bed cultures was compared. The cells cultivated in static solid culture exhibited the highest activity. Possible accumulation of energy sources by the cells was suggested as the explanation for better performance. PMID- 14545673 TI - Caffeine reduction in coffee pulp through silage. AB - Silage tests to study reductions of antiphysiological compounds (caffeine and polyphenols) of fresh coffee pulp during the anaerobic fermentation were done. A concrete silo divided in compartments, with a total capacity of 9 tons of fresh material was utilized. The silage periods ranged between 99-224 days and the following materials were ensiled: 1) coffee pulp, 2) coffee pulp with sugar cane molasses, 3) coffee pulp with a mixture of molasses and ammonia and 4) screw pressed coffee pulp with molasses. Reductions in caffeine, total polyphenols and condensed polyphenols ranged between 13-63%, 28-70% and 51-81% respectively. It was concluded that in the case of coffee pulp, silage presents and ideal method to preserve the material and partially reduce the contents of antiphysiological compounds. PMID- 14545674 TI - Development of high penicillin producing strains for solid state fermentation. AB - Penicillin production with an industrial strain and 4 strains of P. chrysogenum, in solid state fermentation (SSF) and liquid submerged fermentation (LSF), was determined. Their ability to produce the antibiotic in SSF in relation to their capacity to do so in LSF was evaluated. this was done by calculating the ratio PS/PL (production in SSF/production in LSF), which was called relative production. Clones were isolated from each strain and evaluated in a similar way. The strains presented different relative productions (from 1.4 to 2.5). Within the clones, a much wider range of relative productions was observed (0.6 to 16.7). On the other hand, the highest-producing strains in LSF were also the highest producers in SSF. This indicates that the production potential of a strain is an important factor in its production level in SSF. Moreover, the highest penicillin producing ciones (9,500 to 10,500 microg of penicillin/g were generated from high-yielding strains (P2 and ASP-78). However, the higher producing strains (in LSF) showed lower relative performance, suggesting that higher producing strains tend to express less efficiently their potential in SSF. In this study, several overproducing clones, particularly suited for SSF, were obtained by the procedures followed. Production increases of 500 to 600 %, in this culture system, were achieved. PMID- 14545675 TI - Effect of particle size, packing density and agitation on penicillin production in solid state fermentation. AB - The use of a large particle size (14 mm) support (sugar cane bagasse) increased penicillin production by solid state fermentation by 37 %, however this effect was due to a higher sugar concentration in this bagasse fraction. Cultures with closer packing densities (0.35) produced 20 % more penicillin. Agitation did not have a negative effect on production if moisture loss during the operation is restituted. PMID- 14545676 TI - Growth of Candida utilis in solid state fermentation. AB - In this work, the growth of the yeast Candida utilis on different solid substrate (wheat bran) and supports (sugarcane bagasse and Amberlite resin) imbibed with a liquid culture medium was studied. Growth was followed by sugars consumption, carbon dioxide production rate (CDPR) and cell count. The results showed the ability of the yeast to grow on the three solid media with fairly good viability and total dextrose consumption in the case of sugarcane bagasse and Amberlite, and partial consumption of wheat bran sugars. After two or three days of culture, a five hundred fold increase in cell population was observed. PMID- 14545677 TI - Production of Colletotrichum truncatum for use as a mycoherbicide: effects of culture, drying and storage on recovery and efficacy. AB - Colletotrichum truncatum (Schwein.) Andrus and Moore NRRL 13737 (= NRRL 18434) is a fungal plant pathogen which shows promise as a bioherbicide against the troublesome weed Sesbania exaltata (Raf.) Rydb. ex A. W. Hill. Previous studies showed similar amounts of spores were produced/ml of medium in liquid and solid state fermentations. In this study, Colletotrichum truncatum spores were produced in liquid (LC), solid/vermiculite (SV), and solid/perlite-cornmeal-agar (SP). After drying at room temperature with flowing air, SV and SP retained the most viability. Each product was then stored at 4 degrees, 15 degrees, and 25 degrees C. All three products stored at 4 degrees C and SP stored at 15 degrees C retained highest viability. Efficacy based upon assays utilizing equal numbers of viable spores showed SV and SP spores incited more severe disease symptoms than LC spores. PMID- 14545678 TI - Use of solid state fermentation to produce Beauveria bassiana for the biological control of European corn borer. AB - The production process of a new bioinsecticide against european corn borer is described. The entomopathogenic fungus, Beauveria bassiana, is cultivated by Solid State Fermentation (SSF). The culture support chosen, clay microgranules, humidified with optimal nutritive solution, is incubated in optimal conditions during 48 hours, then dried for 5 days. The bioinsecticide can be directly used after harvesting, without formulation. This process is original for several reasons : - The granulometry (500 microm) and the bulk density (0.6) of the microgranules are compatible with the mechanical standard application of pesticides on corn. - The bioinsecticide could be produced in a pilot reactor of 1600 1 capacity, on 0.5m-thick-layer.- The biomass bound to microgranules conserves its efficiency after storage for 12 months at 4 degrees C, unlike pure cells. - Like the chemical insecticides usually used, this bioproduct has a field efficiency of 80%. Moreover, the efficiency of the product persists during 3 weeks. PMID- 14545679 TI - Solid state fermentation reactors: from lab scale to pilot plant. AB - Relatively many workers in the world are studying different aspects in SSF processes but few are working on reactor design and scale-up. From about 10 years, we are developing reactors from lab scale to pilot plant, based on the same technology, reactor design and flowsheet to allow fermentation with a deep layer (up to 1 m in the pilot plant). These reactors have all a forced aeration and the possibility or not to agitate. Regulations of temperature and water content of the culture are monitored by a special device. PMID- 14545680 TI - Operational parameters for packed beds in solid-state cultivation. AB - Packed bed cultivation systems have potential for widespread application in solid state cultivation (SSC), but they are poorly characterized. The effects of particle size and substrate loading on the growth of Rhizopus oligosporus on sago beads in packed bed bioreactors were investigated. Pressure drop and protein were monitored as indicators of fungal growth in cultivations performed in a large column (4.9 cm internal diameter and 60 cm height) and a system of small columns (4.2 cm internal diameter and 5.2 cm height). The differential pressure drop increased to a maximum between 34 and 44 h and then decreased again. The maximum differential pressure drop attained was greatest for the smallest particle size and for the lower substrate loadings. However, since the protein content continued to increase throughout the cultivation, pressure drop could not be used to monitor growth directly. PMID- 14545681 TI - Gas concentration and temperature gradients in a packed bed solid-state fermentor. AB - In solid-state fermentation (SSF), interaction of heat and mass transfer with biochemical reaction (growth associated enzyme production) affects the bioreactor performance. This interaction was earlier observed to cause temperature and gaseous concentration gradients which reduced the effective bed height of the bioreactor. Since forced aeration is known to alleviate this problem, a packed column bioreactor with forced aeration was employed in the present study. Using wheat bran and Aspergillus niger CFTRI 1105, experiments were conducted for the production of the enzyme amyloglucosidase at various air flow rates. Temperatures and gas concentrations were recorded and enzyme activities estimated at different bed heights during the course of SSF. Gas concentration and temperature gradients decreased with increasing air flow rate. The packed column allowed the use of larger bed heights and yielded higher enzyme activities (6,260 Units/gDMB) than trays (345 Units/gDMB). Enzyme activity was affected more by temperature than concentration gradients, and increased with air flow rates. PMID- 14545682 TI - Cellular automata simulations of fungal growth on solid substrates. AB - Growth of filamentous fungi on the surface of cereal grains is a critical aspect of solid substrate fermentation (SSF). Numerous mathematical models have been developed to describe various aspects of fungal growth in SSF. These models consider hyphal geometry and nutrient availability as determinants of colony morphology and fungal physiological state. This work describes the use of cellular automata (CA) as an alternative method of modeling fungal growth. CA models reliant on a very limited set of rules or "knowledge base" display a rich array of behaviors that mimic fungal growth. By incorporating probablistic growth rules into CA models, colony characteristics such as biomass accumulation rate, colony radial growth rate, mycelial density and fungal differentiation are readily generated. PMID- 14545683 TI - Removal of fiber from vines by solid state fermentation/enzymatic degradation: a comparison of flax and kudzu retting. AB - Kinetic data describing the decomposition of the outer sheath of kudzu vines (undergoing a solid fermentation process in a glass beaker of soil) have been analyzed to determine the two constants, K(m)/S(o) and V/S(o), where S(o) is the initial substrate concentration, K(m) the Michaelis constant, and V the maximum product rate. The kinetic data are expressed by a simple time-varying desheathing index, obtained from the number of spatula scrapings required to reach the desired hard cellulosic fibers (vascular bundles) of the plant. A simple relationship between the desheathing index, D.I. and the dimensionless product concentration, P/S(o), is proposed to relate the D.I. data and enzyme kinetic concentration data. Thus, the Michaelis-Menten enzyme kinetic parameters can be estimated from easily obtained physical (non-chemical data; the D.I.(t) measurements). This low energy process for processing vines into valuable fibers is similar to the traditional microbial method for recovering flax fibers for linen cloth, by retting of the flax plant vines, except there is no unbound liquid water present in the soil medium. PMID- 14545684 TI - Biopulping process design and kinetics. AB - Biopulping is the solid-state fermentation of wood chips as a pretreatment for mechanical pulping processes. The two organisms that are currently of the greatest interest for biopulping are the white-rot fungi, Phanerochaete chrysosporium and Ceriporiopsis subvermispora. P. chrysosporium has been shown to successfully biopulp wood (33% energy savings; 39% improvement in tear index) without the need for sterilization of the wood or nutrient supplementation. Demonstrating the practical and economical feasibility of the biopulping process requires process modeling based on accurate kinetic data. Techniques to monitor dry weight loss and growth rate as functions of time using carbon dioxide production data have been developed. Growth was shown to be linear with time on unsupplemented chips and exponential with time on supplemented chips. PMID- 14545685 TI - Multivariate FTIR analysis of substrates for protein, polysaccharide, lipid and microbe content: potential for solid-state fermentations. AB - Components of fermentation processes such as protein, polysaccharide and lipid, as well as microbes, such as fungi grown on solid substrates, are difficult to measure in situ. The potential of Fourier Transform Infrared (FTIR) analysis of solid-state fermentations from mid-infrared absorption spectra has been investigated. The problem under consideration was to build a calibration model containing no irrelevant information to enable a multivariate mathematical approach for prediction of component concentrations. Methods for solid sample preparation and preprocessing of FTIR data were developed to assure Beer-Lambert law compliance and produce a well-conditioned multivariate system. The model was tested using composite samples of zein protein, corn starch and azolectin lipid, and corn samples containing known levels of fungal contamination. Preliminary concentration estimates were remarkably close to the correct values, with less than 5% standard error of prediction for all components measured. PMID- 14545686 TI - Use of chitin measurement to estimate fungal biomass in solid state fermentation. AB - Twenty-two strains of twelve species of Deuteromycotina: Hyphomycetes were studied. Most of them had a variable glucosamine amount (standard deviation higher than 5%). However, if we consider that the amount of glucosamine is constant, the accuracy of the method remains satisfactory (10% instead of 5%, which is the accuracy of the chitin hydrolysis and colorimetric glucosamine measurement). So, by means of this measurement, the fungal growth kinetics could be followed on different solid media (vegetable material such as sugar beet pulp and sponge or mineral like clay granules) used. It is important to note that this method should not be used to compare different media without calibration. PMID- 14545687 TI - Growth estimation of solid-state koji by covering a cellophane membrane on the mash. AB - In a solid-substrate fermentation system, fungal growth within a solid mash is an important index for the efficiency of the saccharification and production of metabolites. Estimation of fungal mass in such a heterogeneous solid-substrate systems is difficult and tedious. In this work, the comparison of Aspergillus oryzae which is a common strain for the wine-brewing process cultured on a cellophane membrane placed on a koji juice agar medium and a small scale of steamed rice koji culture was conduted. Experimental results showed that the cellophane membrane technique resembled the steamed rice koji culture and is considered as a convenient and effective way for investigating the growth characteristics and cytology for solid-substrate koji system. PMID- 14545688 TI - Assessment of solid state fermentation by a biolectronic artificial nose. AB - The use of a bioelectronic artificial nose (AN) is described for on-site monitoring to assess the reaction patterns of solid state fermentations for control of microclimate at sampling site. The nose consists of fractionated olfactory cell proteins of bullfrog (as a receptor membrane) coated on a piezoelectric quartz crystal connected to an oscillator and a data recorder with a personal computer (PC). Five fractions of the olfactory cell proteins gave 5 probes. The response of the nose was analyzed by the PC either as profiles to show the kinds of doors, or as scale factors to show the intensity of odours. Definite differences were noted between the profiles of the headspace gases from earlier and later stages of composing of solid hog wastes. The scale factors showed a single large peak in the earlier stage and one small peak in the later stage. The former showed vigorous biodegradation of organic matter and the latter indicated stabilization of the finished compost. In addition to its use in composing, the nose can discriminate various kinds of alcoholic beverages. PMID- 14545689 TI - The effect of cationic electrolytes on the adhesion of cells. AB - The adhesion rate of cells under charge regulation onto a spherical collector with constant potential is investigated in this paper. Particularly, the effect of the presence of cationic electrolytes in the suspension medium on the adhesion rate is examined. The result reveals that the presence of cationic electrolytes in the suspension medium raises the electrostatic repulsion force between cell and collector surface, when the separation distance between them is small than a critical value. This has the effect of decreasing the adhesion rate of cells. The adhesion rate of cells is quite sensitive to the value of Hamaker constant, especially at a high ionic strength value. PMID- 14545694 TI - The enzymatic hydrolysis and fermentation of pretreated wood substrates. AB - Aspenwood chips were pretreated by steam explosion. The various wood fractions obtained were assayed for their ability to act as substrates for growth and cellulase production of different Trichoderma and Clostridium thermocellum species. Steam exploded aspenwood was as efficiently utilized as solka floc and correspondingly high cellulase activities were detected in the various culture filtrates. When T. harzianum E58 was grown on increasing concentrations of solka floc, highest cellulase and xylanase activities were detected at 1% substrate concentrations while high substrate concentrations (10-20%) inhibited growth and enzyme production. When the cellulosic substrates were supplemented with increasing amounts of glucose, cellulase and xylanase production were inhibited when the glucose concentration exceeded 0.1%. Highest xylanase activities were detected after growth of T. reesei C30 and T. harianum E58 on xylan and solka floc respectively. All of the steam exploded fractions were at least partially hydrolyzed by the T. harzianum E58 cellulase system. The extent of the pretreatment also influenced the ability of Zymomonas mobilis and Saccharomyces cerevisiae to ferment the liberated sugars to ethanol. About 85% of the theoretical yield of ethanol from cellulose could be obtained from the combined hydrolysis and fermentation of pretreated aspenwood. PMID- 14545695 TI - Biochemistry of the oxidation of lignin by Phanerochaete chrysosporium. AB - The objective of this research was to identify the biochemical agents responsible for the oxidative degradation of lignin by the white-rot fungus Phanerochaete chrysosporium. We examined the hypothesis that activated oxygen species are involved, and we also sought the agent in ligninolytic cultures responsible for a specific oxidative degradative reaction in substructure model compounds. Results of studies of the production of activated oxygen species by cultures, of the effect of their removal on ligninolytic activity, and of their action on substructure model compounds support a role for hydrogen peroxide (H(2)O(2)) and possibly superoxide (O(2)(*)(-)) in lignin degradation. Involvement of hydroxyl radical (*OH) or singlet oxygen (1O(2)) is not supported by our data. The actual biochemical agent responsible for one important oxidative C-C bond cleavage reaction in non-phenolic lignin substructure model compounds, and in lignin itself, was found to be an enzyme. The enzyme is extracellular, has a molecular weight of 42,000 daltons, is azide-sensitive, and requires H(2)O(2) for activity. PMID- 14545696 TI - Applications of recombinant DNA technology to the pulp and paper industry. AB - New gene selection techniques (Recombinant DNA) are currently available to exploit useful properties of various biological systems hitherto regarded as interesting but of little or no immediate commercial value. The application of genetic engineering techniques to problems in the Pulp and Paper Industry are many. As a first step these techniques are being used to provide much needed fundamental information on the cellular and molecular mechanisms involved in the expression of extra-cellular enzymes that degrade lignocellulosic pulping wastes. The information gleaned from the studies on cellulolytic fungi and bacteria can be used to genetically engineer a yeast or bacterium capable of converting pulping wastes into ethanol and other useful by-products. PMID- 14545697 TI - Production of useful modified lignin polymers by bioconversion of lignocellulose with Streptomyces. AB - Lignin degrading strains of Streptomyces were grown on lignocelluloses from a variety of plant sources. These actinomycetes readily degraded the lignin present in the residues and released a major portion of the lignin into the growth medium as a water soluble, modified polymer. The polymer, an acid precipitable polyphenolic lignin (APPL), was recovered from spent culture media by acid precipitation or dialysis/lyophilization. APPL's were shown to be mostly free of nonlignin components. As compared to native lignin they were more oxidized, were especially enriched in phenolic hydroxyl groups, and were significantly reduced in methoxyl groups. The yield of APPL from different lignocelluloses correlated with their biodegradability. Grasses such as corn stover were the optimal lignocellulose type for APPL production by Streptomyces. In contrast white-rot fungi produced only small amounts of APPL as they decomposed lignin. A solid state bioconversion process was developed using Streptomyces viridosporus T7A to produce APPL from corn stover lignocellulose in yields >or= 30% of the initial lignin present in the substrate. APPL produced by S. viridosporus was examined for its properties and possible use as an antioxidant. The APPL was shown to have good antioxidant properties after mild chemical treatment to reduce the alpha carbonyl groups present in the APPL. Oxidation of the APPL with hydroxyl radical (OH(*)) further improved its antioxidant properties probably as the result of aromatic ring hydroxylation reactions. As compared with currently used commercial antioxidants, the modified APPL was thought to be competitive when economics of production was considered. Native lignin on the other hand was shown to exhibit no antioxidant properties, even after reduction and/or oxidation. PMID- 14545698 TI - Hydrolysis of xylan and fermentation of xylose to ethanol. AB - The focus in the development of pulping processes has usually been exclusively on cellulose. However, hemicellulose could serve as a valuable source of hexose and pentose sugars. Consequently, it should not be destroyed in a process designed for very high cellulose fibre yields. Novel procedures developed for production of ethanol by the fermentation of pentoses as well as hexoses provide new possibilities of hemicellulose utilization. Many fungi produce extracellular hemicellulases. In the present work the production of xylanase and beta xylosidase with strains of Aspergillus and Trichoderma was studied. The enzymes were used for the hydrolysis of xylan. Xylose was fermented to ethanol by the mold Fusarium oxysporum. PMID- 14545699 TI - Paper pulpmill sludge utilization: techno-economic potential for fuel ethanol, methane and scp production. AB - Various processes have been developed or proposed for converting cellulosic residues from pulp and paper mills into products which can be used for fuel or food. Among the promising practical possibilities are processes for ethanol, methane and microbial protein production by fermentation technology. Given the current Canadian financial climate and product demand, the results of techno economic sensitivity analyses of these three process options indicate that microbial protein production for animal food applications is the most attractive followed by methane then ethanol, the last being quite uneconomical at present. Ironically, research emphasis seems to be placed in the reverse order. It is evident that the relevant costs of upstream and downstream processing in the various process proposals have not been adequately addressed. Case studies of several scenarios illustrate the problems. PMID- 14545700 TI - A continuous biological process to decolorize bleach plant effluents. AB - Although almost every U.S. pulp mill has a biological wastewater treatment system, these systems based on bacteria, are largely ineffective in the removal of color. For this reason, we have attempted to utilize Phanerochaete chrysosporium, a fungus known to degrade lignin, as the primary organism in a novel waste treatment scheme named the MyCoR Process. Color from bleached Kraft mills originates principally from the first extraction stage of the bleach plant. It is this waste stream which is sent to the MyCoR Process reactor, a rotating biological contactor, for decolorization. We have found that under optimal conditions up to 2,000 color units/L/day can be removed from the waste stream. There is also a concomitant removal of COD and BOD. In addition, chlorolignins originating from the bleaching process were found to be dechlorinated; this is of interest to those concerned with the impact of bleach plant effluents on the environment. The process uses conventional wastewater treatment equipment. However, the use of a pure culture of fungus in a secondary metabolic state has not been attempted previously in a waste treatment scheme. Minor equipment modification and close operator attention may therefore be required. A preliminary economic analysis shows that the MyCoR Process, in its present state, would cost about US$30/metric ton of bleached Kraft pulp produced. This cost will decrease as improved or new strains of fungi are developed for the process. PMID- 14545701 TI - Enzymatic control of biological deposits in papermaking. AB - Deposit control in the pulp and paper industry has traditionally been accomplished by the use of toxic biocides. A method has been found whereby biological deposits can be controlled by the use of an enzyme-based product. Numerous field studies have been conducted successfully and photographs prepared illustrating the process. The dynamics of deposit formation and problems associated with such formations have been the subject of considerable study. Development and control of deposit problems under different paper mill conditions using the chemical-biochemical approach will be discussed. PMID- 14545702 TI - The role of microorganisms in the formation of pitch deposits in pulp and paper mills. AB - The cause of pitch deposit formation seems still not fully understood. The work reported here demonstrates that microorganisms effect the agglomeration of emulgated resin droplets and the formation of sticky precipitates. Pitch deposits from mills consist mainly of ethanol-soluble resins. It is also the ethanol soluble fraction of wood resins which forms stable emulsions and which is easily agglomerated by microorganisms. Pitch deposits, collected from various pulp and paper mills, were all found to contain large amounts of microorganisms. Sterile resin emulsions prepared from pitch deposits remained stable over long periods. After inoculation with microorganisms the emulsions were destabilized and the resins completely precipitated as sticky lumps. Various bacteria and fungi are capable of agglomerating the resins, but species isolated from water, pulp and slime in paper mills were usually most effective. Resins from fresh wood were precipitated at a faster rate than aged resins. Problems of pitch formation can be considerably reduced when microbial growth is kept under control in the production system. To be effective, the control measures, e.g. slimicides, must be applied at the right place and time, and in the correct concentrations. This presupposes a thorough knowledge of the plant's microbiological condition which can only be obtained by microbiological examination. Practical cases of the appropriate application of biocides in pulp and paper mill systems are discussed. PMID- 14545703 TI - Anaerobic treatability of waste water from pulp and paper industries. AB - A black liquor evaporator condensate from a Kraft mill and a waste water from production of corrugating medium were anaerobically treated on a laboratory scale. The composition of the waste waters was determined before and after treatment in fixed bed reactors. Toxicity studies by the Microtox-method showed that both waste waters were highly toxic and a slight decrease in toxicity was achieved by anaerobic treatment. Despite the toxicity efficient anaerobic treatment was obtained. Major components of the condensate were methanol, ethanol, acetone, guaiacol, hydrogen sulfide and dimethyl disulfide. Anaerobic treatment reduced the concentration of the major components considerably with one exception. The concentration of hydrogen sulfide was unchanged. Organic overloading of the fixed bed reactor or a temperature drop resulted in an accumulation of acetone, although methanol and ethanol were degraded. Major components of the waste water from the production of corrugating medium were: Klason-lignin, acid-soluble lignin, carbohydrates, extractives and ash. When the fixed bed reactor was operated at a volumetric load of 1.6 kg COD/m(3).d the following reductions were obtained: Klason - lignin (solids fraction 84%; soluble and colloidal fraction 76%), acid-soluble lignin (solids fraction 56%; soluble and colloidal fraction 7%), carbohydrates (100%), extractives (71%), total-S (80%), COD (73%) and BOD(7) (78%). Kinetic studies showed that condensate was more easily degraded anaerobically than corrugating medium waste water. PMID- 14545704 TI - Bacterial degradation of ammonium lignosulfonate. AB - Bacterial strains were selected for their capacity to assimilate and to transform ammonium-lignosulfonate. Modification of the methyl content and of low molecular weight alkyl functions were demonstrated by gas-chromatography and HLPC analysis. Most of these strains completely degraded simple phenolic compounds related to lignosulfonate without inhibition by the carbohydrates present in the liquor. Further investigation suggested that the enzymes involved in the fission of the aromatic nuclei were constitutive in the strains tested. PMID- 14545890 TI - Biotechnology has been associated with the pharmaceutical and brewing industries. PMID- 14545705 TI - Anaerobic fluidised bed for the purification of effluents from chemical and mechanical pulping. AB - Anaerobic treatment has seldom been used for wastewaters from the pulp and paper industry and other branches of the chemical industry. Escape of volatile pollutants to the atmosphere, which always occurs during aerobic treatment, is avoided, and much less sludge is being produced than in an aerobic process. The greatest obstacle for using anaerobic treatment in the pulp and paper industry is the large wastewater volume, which necessitates short hydraulic detention times, because the treatment is to occur in an enclosed space. We used solid carrier particles to prevent wash-out of biomass from the reactor at high hydraulic loading, and an up-flow system in order to be able to use very small carrier particles, maximizing the surface area for biomass attachment. In this paper we describe and discuss the results obtained with this type of anaerobic reactor (fluidised bed) at bench and semitechnical scale for wastewaters from pressurized ground wood pulping and paper manufacture, sulphite pulp evaporator condensate and bleach waste. Earlier work with Kraft pulp bleaching effluent and thermomechanical pulping wastewater and evaporator condensates using anaerobic reactors is also discussed. The results obtained thus far show that there are several wastewater streams from the pulping industry, where 60 to 90% of the dissolved organic pollutants (measured as COD(Cr) or TOC) was biodegraded within 4 to 24 h. The high strength waste streams (COD(Cr) >2000 mg O(2) 1(-1)) allowed organic space load of 4 to 10 kg COD(Cr) m(-3) reactor volume d(-1). With low strength wastes the hydraulic loading was the limiting factor. PMID- 14545892 TI - Anaerobic biological treatment of wastewaters from the pulp and paper industry. PMID- 14545891 TI - Advances in microbial delignification. AB - Microbial delignification is a new field of applied research. The progress will therefore run parallel to the development of new basic knowledge on the physiological demands of white-rot fungi to degrade lignin and on new knowledge on enzyme mechanisms involved in lignin degradation. In the last few years both basic and applied research on microbial conversion of lignocellulosic materials have vastly expanded. In certain areas, such as microbial delignification, considerable progress has recently been made. Basidiospores from Sporotrichum pulverulentum and some CEL(-) mutants have been obtained. Crossing of mycelium from single basidiospore cultures of wild-type and CEL(-) mutants will eventually give rise to much better CEL(-) mutants than those which have been used in the past. An understanding of which enzymes are the most important for lignin degradation to take place is also beginning to develop. This review discusses present knowledge and future possibilities in this field. PMID- 14545894 TI - Genetically engineering mammalian cell lines for increased viability and productivity. AB - The generation of new host cell lines for the production of foreign proteins can be achieved by cell engineering. This approach can be used to enhance the cell's ability to produce proteins that are properly processed and secreted at elevated levels and consequently can increase the overall productivity of an expression system. One potential target for cell engineering is the modification of the cell's protein folding capacity. The appropriate folding, assembly, localization and secretion of newly synthesized proteins is dependent upon the action of a group of proteins known as molecular chaperones. Improving the host cell's chaperoning capacity might increase the yield of properly folded recombinant proteins by preventing the formation of insoluble aggregates. Another potentially beneficial cell engineering goal is the inhibition of physiological cell death. The productivity of genetically engineered cells is dependent upon the maintenance of high levels of cell viability throughout the bioprocess period. Fluctuations in a cell's environment can trigger a deliberate form of cell death known as apoptosis. The proteins that mediate this self-destruction are currently being characterized. Regulating the expression of these death genes by cellular engineering could limit the loss of productivity that results from the physiological death of the recombinant cell line. PMID- 14545895 TI - Plasmid stability and ecological competence in recombinant cultures. AB - The instability of cell cultures containing plasmid vectors is a major problem in the commercial exploitation of molecular cloning techniques. Plasmid stability is influenced by the nature of the host cell, the type of plasmid and/or environmental conditions. Plasmid encoded properties may confer a selective advantage on the host cell but can be an energy drain due to replication and expression. Stability of recombinant cultures ultimately may be determined by the cost to benefit ratio of plasmid carriage. The relative competition between plasmid containing and plasmid-free or indigenous populations can determine the degree of dominance of recombinant cultures. The use of inocula in biotechnological processes in which dynamic environmental conditions dominate may also result in instabilities resulting from the characteristics of the ecosystem. In such dynamic conditions plasmid stability is just one contribution to culture stability. Strategies to enhance plasmid stability, within such environments, based on manipulation of physiological state of host cells, must consider the responsiveness or plasticity of both cells and populations. The robustness of cells or the responses to stresses or transient environmental conditions can influence the levels of instability detected; for example, instability or mutation in the host genome may lead to enhanced plasmid stability. Competition among subpopulations arising from unstable copy number control may determine the levels of recombinant cells in open versus closed fermenter systems. Thus the ecological competence (ability to survive and compete) of recombinant cells in dynamic or transient environments is fundamental to the understanding of the ultimate dominance or survival of such recombinant cultures and may form the basis of a strategy to enhance or control stability either in fermenter systems or dynamic process environments. The creation of microniches in time and/or space can enhance plasmid stability. Transient operation based on defined environmental stresses or perturbations in fermenter systems or in heterogeneous or dynamic environments found in gel immobilized cultures have resulted in enhanced stability. Spatial organization resulting from immobilization has the additional advantage of regulated cell protection within defined microenvironments and controlled release, depending on the nature of the gel, from these microenvironments or microcosms. This regulation of ecological competence allied to the advantages of microbial cell growth in gel microenvironments combined with the spatial organization (or juxtapositioning of cells, selective agents, nutrients, protectants, etc.) possible through immobilization technology offers new strategies to enhance plasmid and culture stability. PMID- 14545896 TI - Applications of modified cyclodextrins. AB - One of the key areas of importance in biotechnology and bioengineering is molecular complexation (MC). MC is useful in selectivity, separation, and solubilization of biomolecules. While many complex, natural MC agents exist, such as proteins and antibodies, relatively few engineered MC materials are available. Inorganic, insoluble MC agents, such as zeolites, are widely used in petroleum catalysis. Carbon Buckminster fullerenes ("bucky balls") can complex small neutral molecules, but are relatively insoluble and difficult to manufacture. Crown ethers have been used for molecular complexation, but are costly to synthesize and have limited capacities. One class of highly useful MC agents are cyclodextrins (CDs). These naturally-occurring, water-soluble cyclic glucans are used in a variety of food, pharmaceutical, and analytical applications. Due to the availability of multiple reactive hydroxyl groups, the functionality of CDs can be greatly increased through chemical modification. A host of new applications are being explored, including enzyme mimicry, molecular recognition, chromatographic separation, and solubilization. This review describes recent applications of modified cyclodextrins in bioprocessing and medicine. PMID- 14545897 TI - Polymerase chain reaction and its applications: special emphasis on its role in embryo sexing. AB - The polymerase chain reaction (PCR) has developed into one of the most promising methods for in vitro enzymatic amplification of DNA and has found widespread application in DNA cloning, sequencing and mutagenesis related studies. This innovative technique can selectively amplify a single target DNA molecule a billion-fold in a span of a few hours. Amplification of specific DNA sequences by PCR is useful in identification of sex, novel genes, pathogens and diseases. PCR has facilitated the establishment of evolutionary relationships among species and in revealing structural intricacies of single cells. In this article we review some of the major advances and applications of PCR, especially, its role in embryo sexing. PMID- 14545898 TI - Advances in microbial biotechnology of bile acids. AB - The recent advances in microbial biotechnology of production of bile acid metabolites helped to identify a number of neutral and acidic steroidal compounds useful as drugs and drug intermediates on a scale which would not have been possible by classical chemical transformations. Microbial transformations viz., hydroxylation, dehydroxylation, reduction of the carbonyl moieties, epimerization, side-chain metabolism, introduction of carbon-carbon double bonds into the steroid nucleus, deconjugation of bile acid conjugates carried out by various microorganisms for production of useful metabolites with special reference to newer techniques including cell immobilization and transposon mutagenesis for selective transformations are reviewed. The different pathways of microbial degradation of bile acids leading to the formation of various products are discussed. A compilation of the metabolites formed by various microorganisms from the bile acids or their conjugates and reported during the period 1979-1992 is also provided. PMID- 14545899 TI - Structure-function relationships in microbial exopolysaccharides. AB - Sufficient well-characterized microbial exopolysaccharides are now available to permit extensive studies on the relationship between their chemical structure and their physical attributes. This is seen even in homopolysaccharides with relatively simple structures but is more marked when greater differences in structure exist, as are found in several heteropolysaccharides. The specific and sometimes unique properties have, in the case of several of these polymers, provided a range of commercial applications. The existence of "families" of structurally related polysaccharides also indicates the specific role played by certain structures and substituents; the characteristics of several of these microbial polysaccharide families will be discussed here. Thus, microbial exopolysaccharides frequently carry acyl groups which may profoundly affect their interactive properties although these groups often have relatively little effect on solution viscosity. Xanthan with or without acylation shows marked differences in synergistic gelling with plant gluco- and galacto-mannans, although the polysaccharides with different acylation patterns show similar viscosity. Similarly "gelrite" from the bacterium originally designated as Auromonas (Pseudomonas)elodea is of greater potential value after deacetylation, when it provides a valuable gelling agent, than it is as a viscosifier in the natural acylated form. The Klebsiella type 54 polysaccharide only forms gels when it, too, has been chemically deacetylated to give a structure equivalent to the Enterobacter XM6 polymer. Both these polysaccharides form gels due to the enhanced interaction with cations following deacylation and to the conformation adopted after removal of the acyl groups. Recent work in our laboratory suggests that deacetylation of certain bacterial alginates also significantly increases ion binding by these polysaccharides, making them more similar in their properties to algal alginates even although the alginates from some Pseudomonas species lack poly-L-guluronic acid sequences. The existence within families of polysaccharides of types in which monosaccharides are altered within a specific structure, or with varying side-chains, also gives an indication of the way in which such substituents affect the physical properties of the polymers in aqueous solution. PMID- 14545900 TI - Cryopreservation of mammalian ova and embryos by vitrification. AB - Vitrification is a new approach to oocyte and embryo cryoconservation. It consists in the solidification of a solution caused not by crystallization, but by a drastic increase in viscosity during cooling. The application of this approach to cryoconservation of oocytes and embryos of different species depends upon the development of proper procedures and non-toxic media. From the technical point of view, the vitrification method is simple and relatively easily applicable under field conditions. The authors review the current procedures applied to oocytes and embryos of laboratory and farm animals. PMID- 14545901 TI - Overproduction of microbial products--facts and ideas. AB - Overproduction of microbial metabolites is related to developmental phases of microorganisms. Inducers, effectors, inhibitors and various signal molecules play a role in different types of overproduction. Primary and secondary metabolism are interconnected. Biosynthesis of enzymes catalyzing metabolic reactions in microbial cells is controlled by well-known positive and negative mechanisms, e.g. induction, repression, catabolite repression, mechanisms controlling enzyme activity include isosteric and allosteric interactions, e.g. competitive and non competitive inhibition, allosteric effects, molecular conversion etc. Biosynthesis of secondary metabolites is catalyzed by unaltered enzymes of primary metabolism, by altered enzymes of primary metabolism and by specific enzymes of secondary metabolism. In addition to classical mutagenesis and selection of suitable microbial cells, methods of molecular genetics are used in the overproduction of microbial products. PMID- 14545902 TI - Genetic designs for product formation in recombinant microbes. AB - Several new bacterial host-vector systems for Klebsiella, Erwinia, Xanthomonas, Nocardia, and Streptomyces have been developed. With these host-vector systems, a strain of Klebsiella, which overproduces the extracellular starch-debranching enzyme, pullulanase, has been developed. The gene for cholesterol oxidase was cloned and used to develop a strain of Streptomyces lividans that extracellularly produces the enzyme, cholesterol oxidase, which is utilized to process cholesterol and diagnostically. The genes for these two enzymes were sequenced, and several interesting facts about their structures and secretory mechanisms were found. For expression of mammalian gene products, the expression vectors. pYM001 to pYM008, containing the lambda P(R)P(L) promoter, which is controlled by a thermolabile repressor, have been developed. The activities of these promoters were compared in various bacterial strains with the galK monitoring system. E. coli promoters, such as lac, trp, tac, lambda P(R), P(L), and P(R)P(L), were found to be expressed in other enteric bacteria and in Bacillus subtilis. With these expression vectors, the vesicular stomatitis virus-nucleocapsid, monkey metallothionein, and human apolipoprotein A1 genes were expressed in E. coli. PMID- 14545903 TI - Overproduction of glutathione and its derivatives by genetically engineered microbial cells. AB - In order to improve the biotechnological potentials of Escherichia coli cells to produce glutathione, S-D-lactoylglutathione and other gamma-glutamyl compounds, the genes for enzymes [gamma-L-glutamyl-L-cysteine synthetase (GSH A) in E. coli B, glutathione synthetase (GSH B) in E. coli B, glyoxalase I (GLO I) in Pseudomonas putida] were cloned and amplified in E. coli. E. coli B cells transformed with both GSH A and GSH B genes exhibited a high activity in the synthesis of glutathione and other gamma-glutamyl compounds in bioreactor systems containing immobilized cells. E. coli C600 cells transformed with GLO I gene of P. putida showed a high GLO I activity and were used for the preparation of S-D lactoylglutathione and other glutathione thiol esters. PMID- 14545904 TI - Threonine production by dihydrodipicolinate synthase-defective mutants of Brevibacterium flavum. AB - A novel type of threonine-producing strains, dihydrodipicolinate synthase (DPS) defective mutants of Brevibacterium flavum, was isolated as alpha-amino-beta hydroxyvaleric acid (AHV)-resistant producers. The third selection markers used were a strong lysine inhibition of threonine production and a lower production of lysine than that of threonine in those derived from strains with feedback sensitive and-resistant aspartokinase (AK), respectively. The maximum threonine production by these DPS-defective mutants was 13.7 g/l at the optimum concentration of DL-diaminopimelic acid (DAP) in a medium containing 100 g/l of glucose, comparable to that by the previously reported conventional producers with feedback-resistant homoserine dehydrogenase (HD(R)). The DPS-defective mutants with feedback-sensitive AK showed a slow but substantial growth in the absence of DAP and their growth was markedly stimulated by DAP, while those with feedback-resistant AK grew well in the absence of DAP and their growth was not promoted by DAP more than that of the parent strain. DPS-defective mutants with HD(R) were derived from an HD(R) mutant producing 10 g/l of L-threonine and selected as AHV-resistant mutants with a higher productivity. The maximum production was 16 g/l. PMID- 14545905 TI - Production of metabolites by methylotrophic yeasts. AB - Recent advances in biotechnology of methanol-utilizing yeasts are briefly summarized. The emphasis is given to production of some fine and commercial chemicals such as formaldehyde, formate, hydrogen peroxide, dihydroxyacetone, ATP, FAD as well as proteins, specifically alcohol oxidase. The advantages of mutants and recombinants derived from methylotrophic yeasts for efficient production of various useful materials are demonstrated. PMID- 14545906 TI - Protein phosphorylation in microorganisms. AB - The covalent modification of proteins by phosphorylation constitutes a major regulatory mechanism. It was first recognized in mammalian tissues. A conclusive evidence for the occurrence of protein phosphorylation and protein kinases in coliform bacteria was obtained in 1978. Several phosphate labeled proteins were found when Salmonella typhimurium was pulse-labeled with 32p(i) and solubilized bacterial contents were analyzed by SDS-polyacrylamide gel electrophoresis. In streptomycetes protein phosphorylation has not yet been demonstrated. We found that Streptomyces albus possesses a protein kinase activity. This in vitro protein phosphorylation is cAMP-independent. PMID- 14545907 TI - Subcellular organization of Streptomyces aureofaciens and overproduction of chlortetracycline. AB - Biosynthesis of chlortetracycline is localized differently under low- and high production conditions (standard low-production strain and its high-production variant). The experimental evidence was based on the assay of anhydrotetracycline oxygenase in subcellular fractions, ultracytochemical localization and electron probe X-ray microanalysis of the product in the mycelium. Overproduction of chlortetracycline is closely associated with compartmentation of biosynthetic enzymes and with an efficient export of the antibiotic out of the cell. PMID- 14545908 TI - Physiology of antibiotic production in actinomycetes and some underlying control mechanisms. AB - Some of the accumulated information on the physiology and nutritional control of antibiotic production in actinomycetes can now be related to recent discoveries in the field of actinomycete molecular biology. This review focuses on aspects of genetic and metabolic control of antibiotic biosynthesis. It surveys some well established principles in the relationship between primary and secondary metabolism, and summarizes briefly the areas where progress is being made in elucidating the molecular organization of regulatory systems underlying this relationship. PMID- 14545909 TI - Biosynthesis and control of beta-lactam antibiotics: the early steps in the "classical" tripeptide pathway. AB - The interaction between growth and secondary metabolism develops from physiological responses of the producer organism to its environment. Nutrients are channelled into primary growth processes or into secondary processes such as antibiotic biosynthesis by a variety of metabolic controls, the nature of which has been extensively studied in organisms producing beta-lactam antibiotics via the tripeptide, delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine. In the following article we review the early stages of beta-lactam biosynthesis in fungi and actinomycetes, keeping in mind the regulation of primary pathways that provide the amino acid precursors of this group of antibiotics, as well as the regulation of the secondary pathway itself. Of special importance to organisms engaging in secondary metabolism are the control mechanisms that suppress the nonessential process during rapid growth but allow secondary metabolic genes to be expressed and resources to be diverted when environmental factors generate the appropriate biochemical signals. PMID- 14545910 TI - Bacterial haloperoxidases and their role in secondary metabolism. AB - Bacteria produce a large number of different halogenated secondary metabolites. Haloperoxidases are believed to be the enzymes responsible for the halogenation reaction. Two classes of haloperoxidases, heme and nonheme, were isolated from different bacteria and their role in the biosynthesis of halogenated secondary metabolites was investigated. Two genes of bacterial haloperoxidases were cloned and can now be used to produce large quantities of the enzymes. PMID- 14545911 TI - Structural and stereochemical aspects of iron transport in fungi. AB - A variety of fungi are known to overproduce and excrete desferri-siderophores under iron limitation. After complexing with ferric iron, octahedral complexes are formed and taken up by siderophore-specific transport systems. These systems represent energy consuming systems as inferred from their sensitivity to respiratory inhibitors, uncouplers and changes of the membrane potential and are able to recognize structure and stereochemical configuration of the various siderophore molecules. Ferrichromes, the most common siderophores in fungi, are generally recognized as Lambda-cis coordination complexes. Triacetylfusarinins, although prevailing as Delta-cis optical isomers in aqueous solution, are assumed to be taken up after isomerization to the corresponding Lambda-cis complexes. However, coprogens which also show a predominant Delta-absolute configuration in solution seem to be transported without prior isomerization. When both, ferrichromes as well as triacetylfusarines or coprogens are taken up, competition during uptake is observed, suggesting the presence of a common transport system during translocation of siderophores across the fungal plasma membrane. PMID- 14545912 TI - State-structure models--a base for efficient control of fermentation processes. AB - The imbedding of state-structure models into the framework of structured segregated models is described. A special type of state-structure models based on delay-differential equations is considered. Some results regarding the description of unbalanced growth processes and the development of efficient periodic control strategies are presented. PMID- 14545913 TI - The mixed substrate concept, applied for microbial syntheses of metabolites. AB - Microbial overproduction of metabolites is a response to suboptimal conditions for growth and multiplication. It is an energy-wasting process in terms of life insofar as a part of energy of the carbon source remains in the metabolite. From an energetic point of view microbial overproduction can be divided into two categories: i) energy-consuming, ii) energy-yielding. The amount of energy required or made available is considered to be responsible for discrepancies between carbon metabolism-determined possible and experimentally obtained yields. Since the expenditure of energy must be provided by oxidation of carbon source more substrate is consumed than required according to the metabolic pathway. In the case of energy-yielding synthesis energy must be discharged. Various possibilities exist. Since metabolic sequences not involved in the synthesis of the proper product are not switched off completely other synthetic processes and even growth can occur. The energy is thus discharged at the expense of substrate. To increase the experimental yield the energy produced or consumed has to be maintained low. This can be achieved by means of substrate mixtures. The synthesis of by-products and growth are difficult to prevent completely. However, growth can be quite desirable since the catalyst is renewed thus making the product synthesis possible. PMID- 14545914 TI - Entropy balances of microbial product formation. AB - Considering the approach of Bermudez and Wagensberg (1986) devoted to the entropy balance of growing microorganisms some equations were developed which describe particularly the entropy balance of microbial product formation. The formula allows to determine the coefficients of resistance R(mn) and of coupling L(mn) according to rates of growth, product formation, maintenance metabolism and heat evolution assuming a linear relationship between thermodynamic fluxes and forces. In order to check the usefulness of the derived model appropriate experimental data of two microbial batch processes concerning production of L-lysine and the antibiotic nourseothricine were taken into account. The results showed similar courses of entropy balances despite different pathways of product formation which were characterized by an overshoot of entropy production at the beginning of biosynthesis of the primary and secondary metabolite. This fact was interpreted as a more general phenomenon for microorganisms under inbalanced nutritional conditions. PMID- 14545916 TI - Biotechnology advances. PMID- 14545915 TI - Achievements in microbial technology. AB - In 1973, recombinant DNA technology was born and the age of the "new biotechnology" came upon us. Today we are seeing the amazing results of recombinant DNA technology, hybridoma technology, enzyme engineering and protein engineering. These techniques are exerting major effects on basic research and on health care, diagnostics and agriculture and soon will bring about changes in other industries such as petroleum, mining, foods and chemicals. Entire pathways of primary and secondary metabolism have been cloned and expressed in foreign microorganisms. The development of recombinant DNA technology is having its major impact on the production of rare polypeptides such as mammalian enzymes, hormones, antibodies and biological response modifiers. In addition to natural polypeptides, analogs are being produced by recombinant DNA technology and this has added an extra dimension of excitement to the field. The future is thus insured for the expanded use of microorganisms in the biotechnological world and the continued improvement in microbial processes to reduce the cost of drugs, enzymes and specialty chemicals. PMID- 14545917 TI - Vaccine design: future possibilities and potentials. AB - Recent developments in the understanding of the structure and replications of a wide range of pathogens, including viruses, bacteria and parasites have opened up ways of designing novel vaccines which should both improve the quality and extend the range and value of vaccines as major prophylactic and therapeutic tools of the future. Two main strategies have emerged, one involving the development of synthetic vaccines which are essentially composed of selected epitopes of the pathogenic agent that will elicit neutralising antibodies. The other strategy attempts to make use of chimeric agents that will allow live virus or bacteria to be used as vectors for carrying appropriate epitopes of the target pathogen. Current knowledge about the immunology and improvements in the presentation of antigen to the immune system will also play an important role in the rational design of vaccines. This review summarises present methods of producing vaccines and considers the development of more rational methods of vaccine design that will greatly influence the production of vaccines in the future. PMID- 14545918 TI - The impact of biotechnology on the dairy industry. AB - The review focusses on the use of genetic techniques to manipulate bacteria that are important to the dairy industry. Both classical and molecular approaches have been used to improve strains involved in yoghurt and cheese production. Examples are provided of methods for; increasing efficiency of substrate conversion, regulating the production of flavour enhancing metabolites, and developing starter cultures resistant to bacteriophage and bacteriocin attack. The possible applications of these systems are discussed. PMID- 14545919 TI - The industrial production of enzymes. AB - The production of enzymes is a pursuit central to the modern biotechnology industry. Markets for traditional industrial enzymes continue to grow while the continued emphasis on biotechnological endeavours has generated demand for an ever increasing number of additional biocatalysts. The advent of genetic engineering has now facilitated the large-scale production of enzymes and other proteins which are produced naturally only in minute quantities. This development is particularly significant with regard to the production of enzymes and other proteins of therapeutic significance, which are now available in clinically useful quantities.The level of downstream processing to which any enzyme is subjected is dependent upon its intended application. Industrial enzymes produced in bulk generally require little downstream processing, and hence are relatively crude preparations. Enzymes destined for therapeutic applications are subject to a far higher degree of downstream processing, often incorporating 3-4 chromatographic steps. While enzymology is one of the longest established branches of the biochemical sciences, it continues to be an area of ongoing, active research. The continual discovery of new enzymes and a greater understanding of previously discovered enzymes and their functional significance suggests many novel applications for these catalytic activities. The intestinal production and utilization of enzymes will continue to be of central importance in the biotechnology industry. PMID- 14545920 TI - Microbial biosorption of metals: potential in the treatment of metal pollution. AB - The phenomenon of metal biosorption by microorganisms has been thoroughly documented. Although this phenomenon is exhibited by both living and non-living forms of biomass, the purpose of this chapter will be to review biosorption by the latter. In addition, the application of various technological processes required for exploitation of this phenomenon in waste treatment will be examined. PMID- 14545921 TI - The use of macrophytes in bioremediation. AB - The development of reedbed technology for bioremediation is reviewed. The future development, potential and implementation of these systems are considered in detail together with the cost effectiveness and ease of maintenance. PMID- 14545922 TI - Gene technology-based antimetabolite design: the use of an in vitro protein expression system to facilitate antimetabolite design for virally-induced human diseases and malignant conditions. AB - A precondition for the chemotherapeutic treatment of a variety of virally-induced human diseases and malignant conditions is a highly selective interaction of the drug molecule to be used with it's biological target. To ensure the development of novel, effective drugs, it is essential that the biological target is well characterised with regard to it's structure and activity. Such characterisation relies upon adequate amounts of pure target being available. One of the most important enzymatic importers for antimetabolites is the enzyme thymidine kinase. In this article an in vitro protein expression system is described which facilitates the production of milligram amounts of pure and biologically active thymidine kinase, from a number of important biological sources. Results have shown that the in vitro produced enzyme has the exact biochemical propeties of the in vivo enzyme. Thus the in vitro protein expression system is an ideal vechicle to facilitate an in depth investigation of the enzyme's biological properties. PMID- 14545923 TI - Nucleic acid probes for the food industry. AB - Recombinant DNA technology has revolutionised microbiological analysis by providing rapid and sensitive methods for the direct detection and identification of organisms. These advances have been most successfully applied to basic research and to clinical microbiology. Continued improvements in the techniques and in assay formats have now brought them to the stage where they could be useful on an industrial scale. The use of nucleic acid techniques for the identification of bacteria of importance to the food industry is discussed with emphasis on developments in practical applications. PMID- 14545924 TI - Commercial exploitation of transgenics. AB - A series of examples of the application of recombinant nucleic acid technology to the production of transgenic organisms is presented. The review considers the distinct advantages of producing certain recombinant biomolecules in appropriate eucaryotic systems rather than in the traditional procaryotic ones. Gene therapy in humans and the production of transgenic animals and plants are discussed. Concerns about the inadvertant construction of new pathogens and ethical considerations about interfering with poorly understood ecosystems are addressed. PMID- 14545925 TI - Live cultures: their use in industrial biotechnology. AB - Advances in the areas of molecular genetics and molecular biology provide a tremendous opportunity for development of custom-made microorganisms for human and animal use. This reviews discusses the current and future potentials of these organisms for use in the food and agricultural industries. PMID- 14545926 TI - A first look at biosensors. AB - This is a very elementary guide to biosensors, outlining the principles of operation of some optoelectronic and current measuring biosensors. Factors which can influence successful translation to the marketplace are addressed. PMID- 14545927 TI - Monoclonal antibody technology: applications in veterinary science. AB - Monoclonal antibodies have revolutionised the study of animals and their diseases. The author looks at the detection of antigen in samples using a range of techniques from indirect fluorescence, through in-situ hybridization to enzyme linked immunosorbent assays. Examples are given of how Salmonella species, mastitis antigens, viral antigens, chlamydial organisms and E. coli toxins can be detected using specific monoclonal antibodies. The recognition of antigen in tissues by monoclonal antibodies is also discussed using as examples; the vitamin biotin, the chicken anemia virus, the growth promoter clenbuterol and the bovine lymphokine, gamma interferon. The ability of monoclonal antibodies to measure specific antibody is also discussed, with particular reference to chicken anemia agent. The review concludes with a discussion of the ability of monoclonal antibody based ELISAs to discriminate between pigs naturally infected with Aujeszky's disease and those vaccinated against the condition. PMID- 14545929 TI - The development of gene expression systems for filamentous fungi. AB - Filamentous fungi have been used for decades in the commercial production of enzymes, antibiotics, and specialty chemicals. Traditionally, improving the yields of these products has involved either mutagenesis and screening or modification of fermentation conditions. Generally, selective breeding of strains has not been successful, because most of the commercially important fungal species lack a sexual cycle. For a few species, strain improvements have been made possible by employing the parasexual cycle for genetic crosses (30). The recent development of DNA-mediated transformation systems for several industrially important fungal species has spawned a flurry of research activity directed toward the development of gene expression systems for these microorganisms. This technology is now a viable means for novel and more directed approaches to improving existing fungal strains which produce enzymes or antibiotics. In addition, fungal expression systems are now being tested for the production of heterologous gene products such as mammalian pharmaceutical proteins. The goal of this review is to present a summary of the gene expression systems which have recently been developed for some filamentous fungi of commercial importance. To insure that the most recent developments are presented we have included data from not only scientific papers, but also from personal communications, abstracts, symposia, and our own laboratory. PMID- 14545930 TI - Expression of proteins encoded by foreign genes in Saccharomyces cerevisiae. AB - The yeast, Saccharomyces cerevisiae is currently used for the production of recombinant DNA-generated proteins derived from a variety of eukaryotic organisms. The applications of a yeast-based technology in the production of proteins for pharmaceutical and industrial purposes is discussed including current methods for introducing recombinant genes into yeast and strategies for maximizing their expression. PMID- 14545931 TI - Development and application of synthetic peptides as vaccines. AB - Vaccination against bacterial and viral diseases has been one of the major achievements in medicine and immunology since the beginning of this century. Extensive vaccination programs have been able to control or, in the case of smallpox, virtually wipe out some of the most dangerous infectious diseases e.g. poliomyelitis, measles, whooping cough, diphtheria and tetanus. However, as this success has been limited mainly to the developed, affluent countries, infectious diseases still remain the worlds largest health problem. Furthermore, vaccines against human parasites are non-existent. Recent advances in immunology and molecular biology including recombinant DNA technology have provided the basis for new approaches to vaccine development. PMID- 14545932 TI - Advances in bioconversion of anthracycline antibiotics. AB - During the last decade new anthracycline-type structures with potential usefulness in cancer treatment have been supplied both by new microbial strains and by bioconversions of precursor molecules employing cells or enzymes. We highlight recent advances in bioconversion of anthracycline structures with the main focus on late transformations such as are carried out by oxidoreductases. PMID- 14546279 TI - Transport regulation of recombinant gene expression in E. coli and B. subtilis. AB - Expression kinetics of the lactose (lac) operon in Escherichia coli are reviewed for both wild-type and recombinant cell cultures under chemostatic conditions. A unified model which involves regulation of active inducer (lactose) transport, promoter-operator regulated expression of the lac operon, glucose-mediated inducer exclusion, and catabolite repression is summarized and supporting data is shown to verify its accuracy. The synthesis of alpha-amylase with a recombinant form of Bacillus subtilis is also reviewed to point out generic features in transport regulation, the lac operon model providing a point of departure. While there are many similarities in the influence of transport on both regulating models, there are also important differences. In a chemostat system, the synthesis of alpha-amylase is nongrowth associated, while beta-galactosidase is a growth-associated enzyme. Nevertheless, transport regulation is an important feature in both instances. PMID- 14546280 TI - Artificial seeds. AB - The use of somatic embryos as artificial seeds for large scale clonal propagation of plants is close to becoming a reality. The quality of the artificial seed depends on the temporal, quantitative and qualitative supply of growth regulator and nutrients along with an optimal physical environment. Desiccation of somatic embryos provides a quiescent phase analogous to true seeds, facilitating the convenience of year round production, storage and distribution. Somatic embryos possess the ability to express desiccation tolerance in response to an external chemical or physical stimuli. The mechanisms of desiccation tolerance involve stabilization of membranes in dry state and prevention of oxidative degradation of biomolecules. Encapsulation of embryo may control the water uptake, release of nutrients and provide mechanical protection required for field planting. PMID- 14546281 TI - Gene transfer strategies in plants. AB - In recent years, a large number of gene transfer methods have been developed. However, the results of these studies have often been published in such a way that it has been extremely difficult for researchers to assess the reliability and efficiency of the method, and to judge whether or not integrative transformation has occurred. Thus although an abundance of knowledge exists within the area of gene transfer, its documentation remains disjointed. This report summarises the recent progress which has been made in the field of gene transfer systems in plants and discusses the associated advantages, disadvantages and limitations in an attempt to clarify this issue. PMID- 14546282 TI - Immunoaffinity purification: basic principles and operational considerations. AB - Immunoaffinity purification has become an important technique in biotechnology. In this review the basic principles of immunoaffinity separations are described with respect to the stages of operation and potential application. The most commonly used support materials, activation procedures, and coupling chemistries are compared to one another for suitability in various applications. Individual operational steps for fixed bed immunoadsorbents including loading, washing, elution and regeneration are described in terms of both theory and practice. Factors influencing adsorbent stability are identified, and alternative operation and configuration strategies are discussed in light of their application to immunoaffinity systems. PMID- 14546639 TI - Microbial cellulose as a specialty chemical. AB - Microbial polysaccharides are extensively used commercially as gelling or suspending agents, as protective colloids or as thickening agents. Until recently, microbial cellulose producing systems such as Acetobacter xylinum, had been used largely as model systems for the study of cellulose biosynthesis. Current advances in molecular biology and biochemical engineering promise to usher microbial cellulose into the specialty chemical market. This review will highlight some of the recent progress made in our understanding of microbial cellulose biochemistry and biosynthesis, describe some of its inherent virtues and identify current unique applications of this versatile biopolymer. PMID- 14546640 TI - Evolving strategies for the genetic engineering of herbicide resistance in plants. AB - Developments in plant genetic engineering technology will shortly permit the commercial introduction of transgenic crop varieties resistant to a number of non selective herbicides. High levels of tolerance have been achieved both by overexpression of a target protein and by modification of that target to an insensitive form. However the results of preliminary trials suggest that in some instances the yield penalty for such genetic alterations will be prohibitive. An alternative strategy, based on the transfer and expression of a gene encoding a herbicide-detoxifying enzyme, appears to offer high resistance levels at low metabolic cost and is expected to assume increasing importance, although it may not prove suitable for all herbicides. PMID- 14546641 TI - Beneficial interactions between micro-organisms and roots. AB - Microbial activity in the rhizosphere can have positive and negative effects on plants. Some of the beneficial processes act by minimizing the negative effects or by modifying the cropping environment to enhance productivity. Processes that are considered here include the provision of nitrogen, phosphorus and iron to the plant, the biocontrol of diseases and deleterious organisms and the stabilization of soil structures. The use of genetic engineering techniques in studying these processes and generating novel strains which may enhance them is discussed, along with the possible consequences of the release of organisms into the environment. PMID- 14546642 TI - Biotechnological approaches to breeding and cultivation of ornamental crop plants. AB - Recent advances in plant biotechnology hold great potential for the ornamental horticulture industry. In addition to conventional methods, breeders can now introduce genetic variation into ornamentals by the application of recombinant DNA technology. This technology is particularly useful for effecting changes in phenotypic expression encoded by single genes such as corolla and foliage color and texture, stem length, scent, temporal regulation of flowering, vase life of cut flowers and resistance to stressful environments. In part, the commercial success of this technology will depend on developing reliable methods of transformation of ornamentals and on the stability of the introduced or altered genes. In addition, new and improved strategies of in vitro culture have been commercially implemented for the propagation and breeding of a wide variety of ornamental crops and will undoubtedly play a major role in the screening and propagation of chimeric plants developed by recombinant DNA technology. PMID- 14547224 TI - Protein engineering. AB - The techniques of protein engineering are proving to be a revolutionary experimental tool for understanding protein structure-function relationships. Even at this early stage, proteins of improved characteristics for specific industrial and therapeutic uses have already been produced. Tailoring enzymatic properties for non-physiological substrate conditions, altering pH optima, changing substrate specificity, and improving stability have already been demonstrated to be feasible. Nevertheless, the ability to make useful proteins which radically differ from a natural structure or designing altogether new structures exceeds present understanding. PMID- 14547225 TI - The development of subunit and synthetic vaccines using recombinant DNA technology. AB - Vaccination of humans and animals against invasion by pathogenic organisms is an effective and integral component of preventive medicine. Traditionally, vaccines have been prepared from various forms of killed or attenuated whole organisms. Such killed or attenuated vaccines presumably retain some of the important antigenic determinants of the organism which can elicit an effective immune response in the vaccinated host. Major drawbacks encountered with these types of vaccines include the introduction of undesirable side-effects after vaccination, as well as induction of only partial protection in some cases. In addition to killed or attenuated vaccines, partially purified antigenic determinants from the whole organism have been used as vaccines. However, the cost and difficulties involved in preparation of the purified antigen often make this an uneconomical approach. Within the last decade, the advent of recombinant DNA technology has brought about a new approach in the preparation of vaccines. In this review, some of the recent developments in several research areas leading to the production of effective vaccines will be presented to demonstrate the promising future of this new approach to vaccine development. PMID- 14547226 TI - Biofilm reactors in anaerobic wastewater treatment. AB - Attached biofilm reactors provide the means for implementing energy-efficient anaerobic wastewater treatment at full scale. Progress has been made in the development of fixed, expanded and fluidized bed anaerobic processes by addressing fundamental reactor design issues. Several new biofilm reactor concepts have evolved from recent studies. PMID- 14547227 TI - Influence of modifying agents on enzyme inactivation studies. An analysis using a series mechanism and a form of the Hill-type equations. AB - A series deactivation model is utilized to theoretically examine the influence of different modifying agents on enzyme deactivation kinetics. A form of the Hill type equation is used to describe the effect of the modifying agents on the model parameters. Modification-induced inactivation equations are presented for the acetylation and succinylation of E. Coli asparaginase, for the site-specific reagent and substrate modification of flavocytochrome b(2) from Baker's yeast, and for the guanidinium chloride inactivation of cathepsin D. The analysis of more data for these and other enzymes would help further substantiate the technique presented and enhance the applicability of the model. PMID- 14547817 TI - Rheology of filamentous fermentations. AB - The performance of a bioreactor containing a filamentous fermentation broth is greatly influenced by the rheological properties of the broth. These properties are determined mainly by the concentration of biomass, its growth rate and morphology. Included in the morphology are such factors as the geometry of hyphae (length, diameter, branching frequency), hyphal flexibility and hyphal-hyphal interactions, which can all be affected by the operational design of the reactor. Thus, correlations describing viscosity as a function of biomass only are of limited value. A better understanding of the relations between morphology and rheology may be achieved by a combination of rheological and morphological studies. Rheological properties are normally determined using off-line measurements in-spite of associated problems with sample treatment influencing the results. Equipment for dynamic, on-line, measurement of morphology and rheology is available, but little used in filamentous fermentations. Controlling the rheological properties of mycelial fermentations may be difficult because of the great number of factors influencing mycelial development and/or hyphal-hyphal interactions.Polymer solutions are often used to simulate flow behaviour of filamentous fermentations and scale-up and mass transfer considerations are based on these studies. Although much information has been gained this way, the predictions developed do not include the effect of an active biomass on the mass transfer and flow properties of the culture. It is important to carry out studies on the non-homogeneous fermentation fluids, and develop correlations based on these studies. PMID- 14547818 TI - Technology and applications for encapsulated spermatozoa. AB - A technique for microencapsulation of bovine spermatozoa has been developed with minimal spermatozoal injury and thus, a potential use in artificial insemination (AI). Membranes made of the following polymers have proven best: poly-l-lysine, polyarginine, polyvinylamine, and protamine sulfate. Successful encapsulation has been achieved for capsules ranging in size from 0.75 to 1.5 mm, and sperm concentrations from 45 to 180 x 10(6) cells/ml. Successful buffers include Cornell University Extender and egg yolk citrate - glycerol (maximum 10% v/v egg yolk for normal capsular shape). Capsule fragility (ability to rupture under aging and physical stress) was negatively related to membrane thickness which ranges from 1.92 microm to 5.32 microm (controlled by polymer concentration, molecular weight, and exposure time) and positively to concentration of sperm encapsulated. On delivery to the porcine reproductive tract, the capsule constructed of poly-l-lysine membranes ruptured between 12 and 24 hr after insemination. Heterospermic studies have shown that encapsulated sperm are capable of fertilization in vivo, but are at a disadvantage to unencapsulated sperm when AI is at conventional times following detection of estrus. PMID- 14547819 TI - Biotechnological applications of research on animal pigmentation. AB - The implications of primary research on pigmentation for the colour manipulation of animal species of economic importance, and the facilitation of specific processes in biotechnology are discussed. Pigment technologists, especially poultry and fish nutritionists, are concerned with achieving the often specific type and degree of coloration demanded by consumers of various products (notably egg yolk, eggshell, broiler skin and salmon flesh). In most instances involving melanin (pelage, plumage and integument) and porphyrin (eggshell) pigments, the desired coloration is achieved through the use of alternate alleles at gene loci controlling the characters of interest. In contrast, coloration involving carotenoids is controlled primarily through pigment supplementation in the diet. The difference between carotenoids and other pigments involves the strict dietary origin of the former. Factors other than pigment availability, such as body condition, hormonal status and genetic constitution, also affect coloration. Although day-old chicks can be sexed by visual inspection of their genitalia, matings resulting in sex-associated phenotypes are in wide use. The genetic markers involved affect the colour of the plumage. The cloning of genes involved in pigmentation offers the prospect of deciphering the genetic control of animal pigmentation and modifying it to meet specific pigmentation needs. PMID- 14547820 TI - Generation and applications of monoclonal antibodies for livestock production. AB - Monoclonal antibodies (MCAs) have found widespread applications in livestock production. Although the generation of murine MCAs is at present a routine, the production of homologous MCAs, especially important for in vivo applications, is still hampered by the lack of efficient homologous fusion partners for immortalization of antibody producing lymphocytes of livestock species. At present, MCAs are used in immunodiagnostic tests e.g. to monitor livestock reproduction and quality of livestock products. In the future MCAs will also be used in immunosensors for real-time and on-site applications in the same areas. The commercial application of MCAs for the immunomodulation of (pharmacologically induced) physiological processes underlying important (re)production traits is at present limited to the use of anti-PMSG MCAs in PMSG-induced superovulation. However, many potentially interesting applications are under investigation (e.g. immunopotentiation of growth hormone to enhance growth; immunocytolysis of adipocytes to increase lean meat production; immunoneutralization of GnRH for immunocastration; immunoimitation of hormone activity with anti-idiotype antibodies). Attempts to use specific MCAs for the sexing of embryos have been disappointing, mainly because of the relatively low accuracy. In the future, MCAs against membrane proteins which are specific for X- or Y-chromosome bearing spermatozoa might be used for bulk separation of livestock sperm. In general, it is expected that engineered (homologous) recombinant MCAs will largely contribute to the development of a new generation of rapid immunodiagnostic tests and effective immunomodulation applications. They will further increase the use of MCAs in livestock production. PMID- 14547821 TI - Renaturation of recombinant proteins produced as inclusion bodies. AB - Expression of recombinant proteins in Escherichia coli often results in the formation of insoluble inclusion bodies. Within the last few years specific methods and strategies have been developed to prepare active proteins from these inclusion bodies. These methods include (i) isolation of inclusion bodies after disintegration of cells by mechanical forces and purification by washing with detergent solutions or low concentrations of denaturant, (ii) solubilization of inclusion bodies with high concentrations of urea or guanidine-hydrochloride in combination with reducing reagents, and (iii) renaturation of the proteins including formation of native disulphide bonds. Renatured and native disulphide bond formation are accomplished by (a) either air oxidation, (b) glutathione reoxidation starting from reduced material, or (c) disulphide interchange starting from mixed disulphides containing peptides. The final yield of renatured proteins can be increased by adding low concentrations of denaturant during renaturation. PMID- 14548467 TI - High density cell culture by membrane-based cell recycle. AB - Enhancement of productivity of a bioprocess necessitates continuous operation of bioreactors with high biomass concentrations than are possible in conventional batch, fedbatch or continuous modes of culture. Membrane-based cell recycle has been effectively used to maintain high cell concentrations in bioreactors. This review compares membranebased cell recycle operation with other such high density cell culture systems as immobilized cell reactors and reactors with cell recycle by centrifugation or gravity sedimentation. A theoretical of production of primary and secondary metabolites in membrane-based recycle systems is presented. Operation of this type of system is discussed with examples from aerobic and anaerobic fermentations. PMID- 14548468 TI - Veterinary vaccines. AB - Vaccination of animals for the prevention of infectious diseases has been practised for a number of years with little change in product composition. Recent advances in molecular biology, pathogenesis and immunology have laid the groundwork for the development of a new generation of veterinary vaccines based on pure subunits as well as live vectored bacteria and viruses. Along with novel methods of antigen preparation, the use of new adjuvants and delivery systems will permit targeting of the appropriate immune response as well as offering flexibility in terms of vaccination protocols. These new technologies are also being applied to the development of vaccines to enhance animal productivity and to control reproduction. PMID- 14548469 TI - Biological safety considerations in the production of health care products from recombinant organisms. AB - Safety considerations in the field of recombinant technology and rDNA production of health care products have been under discussion since the beginning of this technology in 1973 and will certainly go on. However no adverse effects, which could have been attributed to rDNA technology have been observed. On the other hand many life-saving and life-improving drugs have been on the market for many years to the benefit of many patients. New technologies and products thereof often provoke uncertainties about their impact on the environment or society. This article discusses some potential risks in the application of rDNA technology to drugs as well as some benefits for patients, society and environment. PMID- 14548470 TI - Surface display of antibodies. AB - To screen antibody libraries that contain many millions of different clones, a selection system is required with an efficiency comparable to that of the immune system. This can be achieved by displaying antibodies on the surface of microorganisms containing the antibody's gene, analogous to the expression of the IgM antigen receptor on the surface of unactivated B-lymphocytes. Specific clones can then be selected using immobilized antigens. The minor coat protein of filamentous phages, pIII, which initiates the infection of E.coli by binding to their F-pili, and the major coat protein, pVIII, have been used as carriers for displaying antibodies on the phage surface. Recombinant antibodies have also been targeted to the cell surface of bacteria by fusing them with outer membrane components derived from lipoproteins, OmpA and an IgA protease. However, only the pIII system has been routinely used for screening antibody libraries. Here we describe the various antibody surface display systems and the screening of antibody libraries generated from the gene repertoire of lymphocytes and by gene synthesis. Finally, we have made a short comparison of the bacterial production of Fabs versus single chain antibodies (scFv). PMID- 14548734 TI - Field testing of genetically engineered microorganisms. AB - The first approved field releases of microorganisms genetically altered in the laboratory have been initiated in the past several years. While most introductions have been carried out in the United States, several tests have also occurred in the United Kingdom and Australia. Although such releases remain controversial in some areas, these pioneering studies have provided significant insight into the environmental behavior and relative safety of applying these microbes in a well-planned and carefully monitored program. PMID- 14548735 TI - Improvement of Rhizobium inoculants by mutation, genetic engineering and formulation. AB - The use of mutation and genetic engineering techniques have resulted in Rhizobium strains with improved characteristics. The latter approach can provide improvement and new traits not achievable previously. Enhanced commercial Rhizobium inoculants, however, still rely solely on traditional approaches including formulation improvement, mutation, and strain selection. The lack of contribution to the commercial product lines by genetic engineering is mostly due to the time delay and financial burdens cast by the regulatory policies rather than a lack of progress in the application of modern genetic technologies. Such constraints may lessen in time as the public becomes more educated about the technologies and the subsequent political pressure decreases. PMID- 14548736 TI - Non-radioactive techniques for the labelling of nucleic acids. AB - Non-radioactively labelled probes potentially have several advantages over radioactively labelled ones, such as increased stability and reduced hazard. As yet, no non-radioactive methods of labelling are as robust or produce as sensitive a probe as 32P. However, there are many options, some of which use approaches familiar to 32P users (nick translation, random priming, tailing). The majority of methods, whether enzymatic or chemical, direct or indirect, ultimately require the detection of an enzyme by a colourimetric or chemiluminescent substrate. Such detection can be achieved variously by direct visualisation (e.g. of a colourimetrically stained blot), by film, by microtitre plate reader or by CCD camera, depending on the choice of assay format and the degree of quantification required. PMID- 14548737 TI - The use of biotechnology in the conservation of tropical germplasm. AB - The rapid erosion of the genetic diversity of both wild and cultivated plants has recently attracted more and more international concern. As a consequence, germplasm conservation techniques have become accessible to protect the third world's germplasm diversity. Several approaches are used for germplasm conservation and utilization. The evaluation and use of these techniques has to be focussed with one important objective; to preserve with as much integrity as possible, the genetic variability of the selected species. Questions have risen about the best way to preserve genetic diversity of the humid tropics, since it is being recognized that these conditions hold much of the world's gene pool of plants. Seed banks are considered the best system for orthodox seed storage. For many tropical species that cannot support dehydration, tissue culture and cryoconservation are the best alternative. Important considerations regarding the adaptation of modern techniques to tropical species have to be carefully analyzed. Quantitative studies of genetic indices of somaclonal variation, or types or mutant plants can be observed. Consideration of plant germplasm as a base for genetic improvement has come a long way, but much remains to be done, particularly, with the plant diversity that exists in the tropics. All efforts to conserve and use genetic resources will contribute to the benefit of future human generations. PMID- 14548738 TI - Bacterial cell disruption: a key unit operation in the recovery of intracellular products. AB - The need for microbial cell disruption has hindered the large scale production of commercial biotechnological products of intracellular derivation. The intracellular nature of many recombinant products and the potential use of the bacterial storage product, PHB as a commodity thermoplastic have renewed interest in the improvement of this unit operation. This paper provides a review of processes of a mechanical, physical, chemical or biological nature used for cell disruption on both the laboratory and large scale. Applicability of the techniques to large scale operation is discussed. Modification of existing processes is suggested for the reduction of energy requirements and improved process economics. The requirements for the liberation of granular intracellular products such as inclusion bodies and virus-like yeast particles are distinguished from those for the liberation of soluble products, mainly proteinaceous in nature. The integrated nature of the process with both upstream and downstream processes is addressed. Finally, the recent approach of selective liberation of soluble products of interest is reviewed. PMID- 14548739 TI - Applications of microbial surfactants. AB - Biosurfactants are amphiphilic compounds produced by microorganisms that are capable of decreasing surface and interfacial tensions. They are useful in remediation of insoluble organic pollutants in soil and marine environments. There are also a large number of industrial uses for biosurfactants. This paper reviews recent research on applications of microbially-produced surfactants. PMID- 14549121 TI - Functionally-stabilized proteins--a review. AB - The maintenance or stabilization of protein or enzyme function is of vital importance in Biotechnology. Investigations of thermophilic organisms, studies of denaturation and the use of enzymes in organic solvents have each contributed to an understanding of protein stability. Enzymes can reliably and reproducibly be stabilized by variety of means including immobilization, use of additives, chemical modification in solution and protein engineering. Examples of each of these are discussed. With these recent advances it appears that a rational strategy for achieving a particular stabilized enzyme or protein may be within reach. PMID- 14549122 TI - Immobilization of animal cells in fixed bed bioreactors. AB - Immobilization of animal cells has become a highly popular means of achieving high-density animal cell cultures. The advantages of immobilization are that it stabilizes cells in culture and enables long-term culture periods to be achieved. Immobilization increases cell productivity by increasing the usable substrate surface area for anchorage-dependent cells, or by facilitating perfusion of anchorage-independent cells. A method for production of secreted biological products from anchorage-dependent and independent cells is described. The method is based on immobilization of animal cells within the polymeric matrix of polyurethane foam, packed in a fixed bed bioreactor. PMID- 14549123 TI - Insect control with baculoviruses. AB - Baculoviruses have been researched extensively for insect control. Three of their features have been particularly attractive: their host specificity and consequential environmental safety, their virulence in host insects, and their capability for causing disease epizootics. There have been four approaches to their use for insect control: as microbial insecticides for short-term insect population suppression, through seasonal colonization or a recurring "booster shot" for control of more than one pest generation, through introduction establishment where the viral species or strain was not indigenous, and through environmental manipulation to make the ecosystem more favorable for viral epizootics. Actual usage of baculoviruses in pest management has been disappointing, particularly with the microbial insecticide approach, primarily for three reasons: economics, slow speed of kill, and adverse effects of the environment on the viruses. The recombinant-DNA revolution has greatly increased the prospects for baculoviruses in insect pest management. PMID- 14549606 TI - Plasmid promiscuity and chastity and its uses. AB - Bacterial plasmids are obligate and intracellular genetic elements that replicate and are maintained autonomously from the chromosome. They are ubiquitous. Some of them are relatively more promiscuous than others. Plasmid genetic systems that contribute to relative promiscuity or chastity in naturally occurring plasmids are described and discussed. Both the promiscuity and the chastity of plasmid based genetic systems have applications in bacterial molecular genetics, in the production of recombinant DNA products and in the breeding and use of desirable bacteria. The role of these systems in such applications is considered. PMID- 14549607 TI - Microbial interactions with germanium. AB - Germanium is a semi-precious, widely-dispersed, and biologically non-essential metal with considerable potential for application in the fields of electronics, computer engineering and medicine. Research on the physiology and genetics of germanium toxicity and accumulation in microorganisms has received scant attention. An understanding of these aspects is important from both fundamental and applied points of view. This review will examine the diverse range of interactions between germanium and microbial species. PMID- 14549608 TI - Genetic modification of Zymomonas mobilis. AB - The bacterium Zymomonas mobilis is a potentially useful organism for the commercial production of ethanol as it is capable of more than double the rate of alcohol production by yeast. However, industrial application of this bacterium has been restricted in part due to the disadvantages of its limited substrate range (glucose, fructose and sucrose) and by-product formation. Progress in strain improvement and genetic manipulation of this ethanologen is reviewed. Methodologies for gaining reproducible gene transfer in Z. mobilis have recently been developed. Genetic modification has led to its growth on the additional substrates lactose and mannitol. Additionally, a range of by-product negative mutants have also been isolated. Further interest has focused on transfer of Z. mobilis genes to other fermentive organisms in order to gain enhanced product formation. Overall, these genetic approaches should lead to development of novel strains of Z. mobilis and other genera, capable of the use of starch, cellulose and xylan in a manner attractive for industrial ethanol production, besides facilitating over production of products from E. coli strains with enhanced capability to grow at high density. PMID- 14550012 TI - Are hybridomas obsolete? PMID- 14550013 TI - Biosynthesis, characterisation, and design of bacterial exopolysaccharides from lactic acid bacteria. AB - Lactic acid bacteria (LAB) are characterised by their conversion of a large proportion of their carbon feed, fermentable sugars, to lactic acid. However, in addition to lactic acid production, the LAB are able to divert a small proportion of fermentable sugars towards the biosynthesis of exopolysaccharides (EPSs) that are independent of the cell surface and cell wall material. These microbial EPSs when suspended or dissolved in aqueous solution provide thickening and gelling properties, and, as such, there is great interest in using EPSs from food grade microorganisms (such as the LAB that are traditionally used for food fermentations) for use as thickening agents. The current review includes a brief summary of the recent literature describing features of the biosynthetic pathways leading to EPS production. Many aspects of EPS biosynthesis in LAB are still not fully understood and a number of inferences are made regarding the similarity of the pathway to those involved in the synthesis of other cell polysaccharides, e.g., cell wall components. The main body of the review will cover practical aspects concerned with the isolation and characterisation of EPS structures. In the last couple of years, a substantial number of structures have been published and a summary of the common elements of these structures is included as is a suggestion for a system for representing structures. A brief highlight of the attempts that are being made to design 'tailor'-made polysaccharides using genetic modification and control of metabolic flux is presented. PMID- 14550014 TI - Production, purification, characterization, and applications of lipases. AB - Lipases (triacylglycerol acylhydrolases, EC 3.1.1.3) catalyze the hydrolysis and the synthesis of esters formed from glycerol and long-chain fatty acids. Lipases occur widely in nature, but only microbial lipases are commercially significant. The many applications of lipases include speciality organic syntheses, hydrolysis of fats and oils, modification of fats, flavor enhancement in food processing, resolution of racemic mixtures, and chemical analyses. This article discusses the production, recovery, and use of microbial lipases. Issues of enzyme kinetics, thermostability, and bioactivity are addressed. Production of recombinant lipases is detailed. Immobilized preparations of lipases are discussed. In view of the increasing understanding of lipases and their many applications in high-value syntheses and as bulk enzymes, these enzymes are having an increasing impact on bioprocessing. PMID- 14550017 TI - Production of adenovirus vector for gene therapy. AB - The field of gene therapy is rapidly expanding with a major focus on the treatment of cancer. Replication-defective adenoviruses are vectors of choice for delivering corrective genes into human cells. Major efforts are directed to design new generations of adenoviral vectors that feature reduced immunogenicity and improved targeting ability. However, the production of adenoviral vectors for gene therapy applications faces a number of challenges that limit the availability of high quality material at the early stages of research and development in the gene therapy field. Moreover, very few papers have been published on the subject and information on large-scale production methods are only available through specialized conference proceedings. This review outlines the problems associated with mass production of adenovirus vectors and describes research efforts by a number of groups who have contributed to optimize production methods. Better understanding of the adenovirus infection and replication kinetics as well as better understanding of complementing cell line physiology and metabolism greatly contributed to improving vector titers and volumetric productivity at higher cell densities. Also, the critical aspect of viral vector quantitation is discussed. PMID- 14550018 TI - Recovery of microalgal biomass and metabolites: process options and economics. AB - Commercial production of intracellular microalgal metabolites requires the following: (1) large-scale monoseptic production of the appropriate microalgal biomass; (2) recovery of the biomass from a relatively dilute broth; (3) extraction of the metabolite from the biomass; and (4) purification of the crude extract. This review examines the options available for recovery of the biomass and the intracellular metabolites from the biomass. Economics of monoseptic production of microalgae in photobioreactors and the downstream recovery of metabolites are discussed using eicosapentaenoic acid (EPA) recovery as a representative case study. PMID- 14550019 TI - Emerging biological materials through molecular self-assembly. AB - Understanding of new materials at the molecular level has become increasingly critical for a new generation of nanomaterials for nanotechnology, namely, the design, synthesis and fabrication of nanodevices at the molecular scale. New technology through molecular self-assembly as a fabrication tool will become tremendously important in the coming decades. Basic engineering principles for microfabrication can be learned by understanding the molecular self-assembly phenomena. Self-assembly phenomenon is ubiquitous in nature. The key elements in molecular self-assembly are chemical complementarity and structural compatibility through noncovalent interactions. We have defined the path to understand these principles. Numerous self-assembling systems have been developed ranging from models to the study of protein folding and protein conformational diseases, to molecular electronics, surface engineering, and nanotechnology. Several distinctive types of self-assembling peptide systems have been developed. Type I, "molecular Lego" forms a hydrogel scaffold for tissue engineering; Type II, "molecular switch" as a molecular actuator; Type III, "molecular hook" and "molecular velcro" for surface engineering; Type IV, peptide nanotubes and nanovesicles, or "molecular capsule" for protein and gene deliveries and Type V, "molecular cavity" for biomineralization. These self-assembling peptide systems are simple, versatile and easy to produce. These self-assembly systems represent a significant advance in the molecular engineering for diverse technological innovations. PMID- 14550020 TI - Biotechnological applications of cyclodextrins. AB - Cyclodextrins (CDs) are a family of cyclic oligosaccharides that are composed of alpha-1,4-linked glucopyranose subunits. Cyclodextrins are produced from starch by enzymatic degradation. These macrocyclic carbohydrates with apolar internal cavities can form complexes with and solubilize many normally water-insoluble compounds. This review describes recent applications of CDs in pharmaceuticals with a major emphasis on drug delivery systems. The utility of these water soluble cyclic glucans in a variety of foods, flavors cosmetics, packaging and textiles is elaborated. The role of these compounds in biocatalysis is also discussed. Cyclodextrins are used in separation science because they have been shown to discriminate between positional isomers, functional groups, homologues and enantiomers. This property makes them a useful agent for a wide variety of separations. PMID- 14550021 TI - Microarray technology: an array of applications that is far from micro. PMID- 14550022 TI - Developmental biology: an array of new possibilities. AB - Microarrays offer biologists comprehensive and powerful tools to analyze the involvement of genes in developmental processes at an unprecedented scale. Microarrays that employ defined sequences will permit us to elucidate genetic relationships and responses, while those that employ undefined DNA sequences (ESTs, cDNA, or genomic libraries) will help us to discover new genes, relate them to documented gene networks, and examine the way in which genes (and the process that they themselves control) are regulated. With access to broad new avenues of research come strategic and logistical headaches, most of which are embodied in the reams of data that are created over the course of an experiment. The solutions to these problems have provided interesting computational tools, which will allow us to compile huge data sets and to construct a genome-wide view of development. We are on the threshold of a new vista of possibilities where we might consider in comprehensive and yet specific detail, for example, the degree to which diverse organisms utilize similar genetic networks to achieve similar ends. PMID- 14550023 TI - Applications of DNA and protein microarrays in comparative physiology. AB - DNA microarrays have revolutionized gene expression studies and made large-scale parallel measurement of whole genome expression a feasible technique in model species where genomes are well characterized. Such studies are perfectly suited to unraveling the complex regulation and/or interaction of both genes and proteins likely involved in most physiological processes. Gene expression profiles are currently being used to identify genes underlying a range of physiological responses. Characterization of these genes will help to elucidate the pathways and processes regulating physiological processes. Expanding the use of DNA microarrays to non-model species that have been critical in elucidating certain physiological pathways will be valuable in determining the genes associated with these processes. Approaches that do not require complete genome information have recently been applied to "non-model" organisms. As whole genomes are sequenced for non-model organisms, the application of DNA microarrays to comparative physiology will expand even further. The recent development of protein microarrays will be critical in understanding the regulation of physiological processes not accounted for at the genomic level. Together, DNA and protein microarrays provide the most thorough and efficient method of understanding the molecular basis of physiological processes to date. In turn, classical physiological approaches will be vital in characterizing and verifying the function of the novel genes identified by microarray experiments. Ultimately, DNA and protein microarray expression profiles may be used to predict physiological responses. PMID- 14550024 TI - DNA microarrays and toxicogenomics: applications for ecotoxicology? AB - Toxicogenomics attempts to define how the regulation and expression of genes mediate the toxicological effects associated with exposure to a chemical. DNA microarrays are rapidly becoming one of the tools of choice for large-scale toxicogenomic studies. An approach in modern toxicogenomics has been to classify toxicity based on gene transcriptional patterns; comparing the transcriptional responses of a chemical with unknown toxicity to those for which the transcriptional profiles and toxicological endpoints have been well characterized. Recent evidence suggests that gene expression microarrays may be instrumental in defining mechanisms of action of toxicants. However, several assumptions are inherent to a toxicogenomic-based approach in toxicology, many of which remain to be validated. Gene expression profiling using DNA microarrays represents a snapshot of the gene transcriptional responses occurring at a particular time and within a particular tissue. Toxicity, on the other hand, represents a continuum of possible effects governed by both temporal and spatial factors that are inextricably contingent upon the exposure conditions. The perceived toxicological properties of any chemical are dependent on the route, dose, and duration of the exposure, and as such, gene expression patterns are also subject to these variables. Correct interpretation of DNA microarray data for the assessment of the toxicological properties of chemicals will require that temporal and spatial gene expression profiles be accounted for. These considerations are further compounded in ecotoxicological studies, during which altered gene expression patterns induced from exposure to an anthropogenic substance must be discernible over and above the complex effects that phenotypic, genotypic, and environmental variables have on gene expression. To this end, the greatest utility of DNA microarrays in the field of ecotoxicology may be in predicting the toxicological modes of action of anthropogenic substances on host physiology, particularly in non-model organisms. Predictable and accurate assessment of the impacts of a chemical substance in ecotoxicology will require that classical toxicological endpoints be used to validate any effects predicted based on gene expression profiling. Validated expression profiling may subsequently find utility in ecotoxicological-based computer simulation models, such as the Biotic Ligand Model (BLM), in which gene expression information may be integrated with geochemical, pharmacokinetic, and physiological data to accurately assess and predict toxicity of metals to aquatic organisms. PMID- 14550028 TI - Cellulose-binding domains: biotechnological applications. AB - Many researchers have acknowledged the fact that there exists an immense potential for the application of the cellulose-binding domains (CBDs) in the field of biotechnology. This becomes apparent when the phrase "cellulose-binding domain" is used as the key word for a computerized patent search; more then 150 hits are retrieved. Cellulose is an ideal matrix for large-scale affinity purification procedures. This chemically inert matrix has excellent physical properties as well as low affinity for nonspecific protein binding. It is available in a diverse range of forms and sizes, is pharmaceutically safe, and relatively inexpensive. Present studies into the application of CBDs in industry have established that they can be applied in the modification of physical and chemical properties of composite materials and the development of modified materials with improved properties. In agro-biotechnology, CBDs can be used to modify polysaccharide materials both in vivo and in vitro. The CBDs exert nonhydrolytic fiber disruption on cellulose-containing materials. The potential applications of "CBD technology" range from modulating the architecture of individual cells to the modification of an entire organism. Expressing these genes under specific promoters and using appropriate trafficking signals, can be used to alter the nutritional value and texture of agricultural crops and their final products. PMID- 14550030 TI - DNA shuffling: induced molecular breeding to produce new generation long-lasting vaccines. AB - The paradigm for classic vaccines has been to mimic natural infection, and their success relies mostly on the induction of neutralizing antibodies followed by long-lasting immunity. The outcome of aggressive chronic infections such as HIV and HCV, the reappearance of fastidious diseases such as tuberculosis and the progression of cancer growth suggest that natural immune responses are definitely insufficient in many cases. A new paradigm is needed to design and develop a new high-efficiency generation of vaccines ideally able to surpass the capabilities of natural immune responses. In vitro evolution is a new, important laboratory method to evolve molecules with desired properties, which appears as an appealing alternative to achieve this goal. In its battle against disease, the vertebrate immune system triggers a series of well-known molecular events in order to produce protective neutralizing antibodies. This natural in vivo response shares remarkable similarities with the in vitro technique known as molecular breeding or "DNA shuffling." This method exploits the recombination between genes to dramatically accelerate the rate at which genes can be evolved under selection pressure in the laboratory, producing optimized high-efficiency mutant proteins. Since new generation vaccines are aimed to overcome natural selection and environmental pressures to fully inactivate rapidly developing pathogen variants, they could be engineered, developed and selected through the application of directed DNA shuffling procedures. This review highlights the potential of the procedure in the complex context of natural immune responses and the equilibrium and interaction existing in nature between hosts and pathogens. PMID- 14550029 TI - Fulfilling the promise of biotechnology. AB - Genetic engineering has produced pharmaceuticals, disease-resistant plants, cloned animals and research and industrial products. While the comparably mature field of medical biotechnology now reveals its true potential, marine biotechnology is still in the realm of the future. As we explore the earth for new sources of natural chemicals, we now search the waters. Myriad organisms, most unknown to us, live there. Many produce compounds that can be commercialized, or the organisms themselves may be commercialized, through genetic engineering methods. For decades, scientists studied the ocean depths searching for unique molecules and organisms. But not until the early 1980s was there a synthesis uniting marine natural products, ecology, aquaculture and bioremediation research under the heading of marine biotechnology. As harvesting enough products from marine sources to produce sufficient amounts, even for study, is nearly impossible, we need to use genomics techniques to identify biologically active compounds. As we damage our oceanic ecosystems through pollution, overfishing and destructive fishing methods, opportunities to learn more about marine organisms and their commercial potential may be limited. Although governments and intergovernmental agencies are committed to funding and expanding oceanic research, more funding is needed to discover and study the ocean's vast, unplumbed resources. PMID- 14550031 TI - Developments and trends in enzyme catalysis in nonconventional media. AB - The conventional notion that enzymes are only active in aqueous media has long been discarded, thanks to the numerous studies documenting enzyme activities in nonaqueous media, including pure organic solvents and supercritical fluids. Enzymatic reactions in nonaqueous solvents offer new possibilities for producing useful chemicals (emulsifiers, surfactants, wax esters, chiral drug molecules, biopolymers, peptides and proteins, modified fats and oils, structured lipids and flavor esters). The use of enzymes in both macro- and microaqueous systems has been investigated especially intensively in the last two decades. Although enzymes exhibit considerable activity in nonaqueous media, the activity is low compared to that in water. This observation has led to numerous studies to modify enzymes for specific purposes by various means including protein engineering. This review covers the historical developments, major technological advances and recent trends of enzyme catalysis in nonconventional media. A brief description of different classes of enzymes and their use in industry is provided with representative examples. Recent trends including use of novel solvent systems, role of water activity, stability issues, medium and biocatalyst engineering aspects have been discussed with examples. Special attention is given to protein engineering and directed evolution. PMID- 14550035 TI - Microbial DNA diagnostic technology. AB - The principle objectives when creating a robust DNA diagnostic assay system are sensitivity, specificity and minimal read-time. To meet these ends, depending on the specifically defined test, various aspects of molecular hybridization methodology must be optimized. In particular, among other things, attention has focused on (i) formulating highly specific probes; (ii) devising sensitive nonisotopic detection systems, (iii) minimizing the extent of preparing clinical samples for assaying, (iv) amplifying the target sequence to augment sensitivity and (v) enhancing hybridization kinetics to speed up the reaction period. In this article, some recent studies that are directed to the development of nucleic acid hybridization systems for clinical diagnosis of microorganisms are considered. PMID- 14550036 TI - Bacillus thuringiensis insecticidal crystal toxins: gene structure and mode of action. AB - Thanks to the techniques of recombinant DNA, there is now abundant sequence information on several endotoxin genes of Bacillus thuringiensis. The task of correlating this sequence information with the economically important aspects of the toxins such as insect specificity, LD(50) and speed of kill is now under worldwide investigation. Progress has also been made on understanding the mechanism of action of the toxins and on identifying the parts of the protoxin which are important in toxicity. Taken together, the mechanistic data and the sequence information allow the first attempts at rational design of mutant endotoxin genes and greatly facilitate the transfer of those genes to other organisms such as plants. More information is still needed, however, as to the nature of the binding site of the toxin and on the three-dimensional structure of the activated toxins. PMID- 14550037 TI - The potential of microalgal biotechnology: a review of production and uses of microalgae. AB - An overview of the various aspects, promises and limitations of microalgal biotechnology is presented. The factors of importance in microalgal cultivation as well as the culture systems are briefly described. Microalgal biomasses can fulfil the nutritional requirements of aquatic larvae and organisms. The biochemical composition of algae can be improved by the manipulation of culture conditions. The nutritive value of the microalgal biomasses for human and animal consumption is also commented upon as well as some socio-economical aspects. Among the sources of required nutrients (N, P), wastewaters and manures can upgraded as culture media for microalgae the safety of which has to be evaluated. Harvesting of the biomass is one of the bottlenecks. The various techniques, physical, physico-chemical and biological are outlined and their feasibility and economic interest examined. Microalgal biomasses can be submitted to various technological transformations. Various processes are reviewed in the light of their effects on safety and nutritional value. The possible extraction of fine chemicals and the preparation of protein concentrates is also reported on. The various uses of microalgae lead to a possible competition, to be evaluated, between systems for the production of food, energy and chemicals. The review finally covers the application of genetic manipulation to microalgae. PMID- 14550254 TI - Endothelial nitric oxide synthase: a determinant of TNFalpha production by human monocytes/macrophages. AB - Until recently endothelial nitric oxide synthase (eNOS) has been associated exclusively with physiological functions, particularly in the cardiovascular system. However, increasing evidence has been accumulated that supports the concept for a role of eNOS in pathophysiology. In particular, detection of eNOS protein and activity in human monocytes/macrophages suggest an immunomodulatory role of this enzyme. Here, we review data that promote the hypothesis that by enhancing TNFalpha production, eNOS activity should be regarded as a novel pro inflammatory parameter in human monocytes/macrophages. PMID- 14550255 TI - Cholecystokinin synthesizes and secretes leptin in isolated canine gastric chief cells. AB - It is well recognized that a product of obese (ob) locus and body weight control hormone, leptin, acts on both short-term satiety for meal-induced termination of food intake (gastric phase) and long-term satiety for energy expenditure via the hypothalamus. The considerable sources of leptin are chief cells for gastric phase and adipocytes for the long-term satiety. The objective of this study was to demonstrate if CCK enhances leptin synthesis and secretion in isolated canine gastric chief cells. Confocal immunofluorescence studies showed that the CCK-A receptor and leptin were colocalized in the endoplasm. Western blotting demonstrated that canine chief cells expressed the leptin peptide and its protein level was enhanced by CCK treatment. An ELISA further showed that CCK dose dependently secreted leptin from isolated canine chief cells. This was reproduced by the high-affinity CCK-A receptor agonist, CCK-OPE. These results indicate that canine chief cells synthesize and secrete leptin in response to CCK via the high affinity state of the CCK-A receptor. PMID- 14550256 TI - A Rho GTPase controls the rate of protein synthesis in the sea urchin egg. AB - Fertilization of the sea urchin egg triggers a Ca(2+)-dependent cortical granule exocytosis and cytoskeletal reorganization, both of which are accompanied by an accelerated protein synthesis. The signaling mechanisms leading to these events are not completely understood. The possible role of Rho GTPases in sea urchin egg activation was studied using the Clostridium botulinum C3 exotoxin, which specifically ADP-ribosylates Rho proteins and inactivates them. We observed that incubation of eggs with C3 resulted in in situ ADP-ribosylation of Rho. Following fertilization, C3-treated eggs were capable of performing cortical granule exocytosis but not the first cytokinesis. C3 caused in both unfertilized eggs and early embryos alterations in the state of actin polymerization and inhibition of the spindle formation. Moreover, C3 diminished markedly the rate of protein synthesis. These findings suggested that Rho is involved in regulating the acceleration of protein synthesis that accompanies the egg activation by sperm. PMID- 14550258 TI - The effect of temperature on adrenergic receptors of alveolar type II cells of a heterothermic marsupial. AB - Fat-tailed dunnarts, Sminthopsis crassicaudata, survive dramatic changes in body temperature during torpor without experiencing surfactant dysfunction. Adrenergic factors regulate surfactant secretion through beta(2)-adrenergic receptors on alveolar type II cells. Temperature has no effect on the secretory response of dunnart type II cells to adrenergic stimulation. We hypothesise that during torpor, dunnart type II cells up-regulate the number of adrenergic receptors present on type II cells to enable stimulation at lower concentrations of agonist. Here, we isolated type II cells from warm-active (35 degrees C) and torpid (15 degrees C) dunnarts and examined the effects of an in vitro temperature change on the number and activity of adrenergic receptors. Torpor did not affect the beta-adrenergic receptor number. However, we observed a significant decrease in adrenergic receptor number when cells from warm-active animals were incubated at 15 degrees C and when cells from torpid animals were incubated at 37 degrees C. cAMP production was significantly higher in type II cells from torpid dunnarts than warm-active dunnarts and this may contribute, in part, to the temperature insensitivity we have previously observed in the adrenergic regulation of surfactant secretion. PMID- 14550257 TI - PI3-kinase and MAP-kinase signaling cascades in AILIM/ICOS- and CD28-costimulated T-cells have distinct functions between cell proliferation and IL-10 production. AB - Both AILIM/ICOS and CD28 provide positive costimulatory signals for T-cell activation, resulting in proliferation and cytokine production. In this study, we attempted to clarify the key signaling molecules in T-cell proliferation, and also IL-2 and IL-10 production, during T-cell activation by CD3 induced by costimulation with either AILIM/ICOS or CD28. We examined the role of both the PI3-kinase/Akt pathway and MAP kinase family members such as ERK1/2, JNK, and p38 kinase in this process. PI3-kinase and Erk1/2 were shown to potentially regulate primary T-cell activation and subsequent proliferation via both AILIM/ICOS- or CD28-mediated costimulation and the Erk signaling cascade was essential for this proliferation induction and also for IL-2 production. The JAK inhibitor, AG490, inhibited this induction. Our studies indicate that IL-2 is necessary for induction of T-cell proliferation and that the quantities of IL-2 produced by AILIM/ICOS ligation are also sufficient for T-cells to proliferate. In contrast, inhibition of Akt and p38, that are phosphorylated by both AILIM/ICOS and CD28 ligation, could downregulate IL-10 production but not T-cell proliferation. These data raise the interesting possibility that the signaling cascades between T-cell proliferation and IL-10 production are regulated by different molecules in AILIM/ICOS- and CD28-costimulated T-cells. PMID- 14550259 TI - Modulation of delayed rectifier potassium current by angiotensin II in CATH.a cells. AB - Angiotensin II (Ang II) modulates, via Ang II type 1 (AT(1)) receptors, the activity of brain catecholaminergic neurons. Here we utilized catecholaminergic CATH.a cells to define the effects of Ang II on delayed rectifier K(+) current (I(Kv)), one of the factors that determines changes in neuronal activation. Receptor binding analyses demonstrated the presence of AT(1) receptors in CATH.a cells. Whole cell voltage clamp experiments in these cells revealed that Ang II (100nM) produced a significant inhibition of I(Kv), that was abolished by the AT(1) receptor blocker, losartan (1 microM), or by inhibition of phospholipase C (PLC) with U73122 (10 microM). Furthermore, this action of Ang II was completely abolished by co-inhibition of protein kinase C (PKC) and calcium/calmodulin protein kinase II (CaMKII). These results demonstrate that Ang II produces an inhibition of I(Kv) in CATH.a cells, via an intracellular pathway that includes PLC, PKC, and CaMKII. PMID- 14550260 TI - Suppression of Helicobacter pylori-induced gastritis by green tea extract in Mongolian gerbils. AB - Since urease of Helicobacter pylori is essential for its colonization, we focused attention on foodstuffs which inhibit the activity of this enzyme. Among plant derived 77 foodstuff samples tested, some tea and rosemary extracts were found to clearly inhibit H. pylori urease in vitro. In particular, green tea extract (GTE) showed the strongest inhibition of H. pylori urease, with an IC(50) value of 13 microg/ml. Active principles were identified to be catechins, the hydroxyl group of 5(')-position appearing important for urease inhibition. Furthermore, when H. pylori-inoculated Mongolian gerbils were given GTE in drinking water at the concentrations of 500, 1000, and 2000 ppm for 6 weeks, gastritis and the prevalence of H. pylori-infected animals were suppressed in a dose-dependent manner. Since the acquisition by H. pylori of resistance to antibiotics has become a serious problem, tea and tea catechins may be very safe resources to control H. pylori-associated gastroduodenal diseases. PMID- 14550262 TI - Tumor necrosis factor attenuates prion protein-deficient neuronal cell death by increases in anti-apoptotic Bcl-2 family proteins. AB - Prion protein gene (Prnp)-deficient(Prnp(-/-)) neuronal cells are more susceptible to serum deprivation compared to Prnp(+/+) neuronal cells. However, little is known about the cell death of Prnp(-/-) neuronal cells under serum deprivation. In this study, as a known neuroprotective agent we analyzed the effect of tumor necrosis factor-alpha (TNF-alpha) on the cell death of Prnp(-/-) neuronal cells. Although expression of Bcl-2 and Bcl-x(L) decreased in a time dependent manner under serum deprivation, treatment with TNF-alpha protected Prnp(-/-) neuronal cells from serum deprivation with an increase in anti apoptotic proteins Bcl-2 and Bcl-x(L). Nuclear morphological analysis using fluorescence microscopy and flow cytometry analysis showed that gene transfer of bcl-2 or bcl-x(L) significantly inhibited apoptosis induced by serum deprivation. These findings indicate that TNF-alpha attenuated cell death of Prnp(-/-) neuronal cells by induction of Bcl-2 and Bcl-x(L),and that decreases in Bcl-2 and Bcl-x(L) played crucial roles in the apoptosis of Prnp(-/-) neuronal cells. PMID- 14550261 TI - Identification of a nuclear protein that promotes NF-kappaB activation. AB - Receptor-interacting protein (RIP) is a serine/threonine protein kinase that is critically involved in tumor necrosis factor receptor-1 (TNF-R1)-induced NF kappaB activation. In a yeast two-hybrid screening for potential RIP-interacting proteins, we identified a novel protein designated as NKAP. Although NKAP interacts with RIP in yeast, NKAP does not interact with RIP in mammalian cells in co-immunoprecipitation experiments. When overexpressed in 293 cells, NKAP activated NF-kappaB in a dose-dependent manner. Moreover, down-regulation of NKAP by antisense RNA significantly inhibited TNF- and IL-1-induced NF-kappaB activation. Immunofluorescent staining indicated that NKAP was localized in the nucleus. Our findings suggest that NKAP is a novel nuclear regulator of TNF- and IL-1-induced NF-kappaB activation. PMID- 14550263 TI - Intracellular delivery of antibodies using TAT fusion protein A. AB - Internalization of antibodies into mammalian cells is a useful method for analyzing and regulating cellular function. In this study, we developed a novel method for the delivery of antibodies into cells using the TAT-fused protein. This fusion protein consists of two functional domains, the protein transduction domain of HIV-1 TAT and the B domain of staphylococcal protein A (SpA), which has an ability to bind to the IgG. The TAT-SpA fusion protein was mixed with fluorescence-labeled rabbit IgG and added to cells. The internalization of antibody was analyzed using confocal microscopy and flow cytometry in living cells. As a result, fluorescence-labeled IgG with the TAT-SpA fusion protein was observed intracellularly. Flow cytometry results demonstrated time course and dose dependence relationships of antibody internalization. These results suggest that the TAT-SpA fusion protein can be a useful reagent for the delivery of antibody into cells. PMID- 14550264 TI - Rapid proteasomal degradation of translocation-deficient UDP glucuronosyltransferase 1A1 proteins in patients with Crigler-Najjar type II. AB - UDP-glucuronosyltransferase form 1A1 (UGT1A1) is the only bilirubin glucuronidating isoform of this protein, and genetic deficiencies of UGT1A1 cause Crigler-Najjar syndrome, a disorder resulting from nonhemolytic unconjugated hyperbilirubinemia. Here we have focused on the instability of a translocation deficient UGT1A1 protein, which has been found in patients with Crigler-Najjar type II, to elucidate the molecular basis underlying the deficiency in glucuronidation of bilirubin. A substitution of leucine to arginine at position 15 (L15R/1A1) is predicted to disrupt the hydrophobic core of the signal peptide of UGT1A1. L15R/1A1 was synthesized in similar amounts to wild-type UGT1A1 protein (WT/1A1) in transfected COS cells. However, L15R/1A1 did not translocate across the endoplasmic reticulum membrane and was degraded rapidly with a half life of about 50min, in contrast to the much longer half-life of about 12.8h for WT/1A1. Our findings demonstrate that L15R/1A1 was rapidly degraded by the proteasome owing to its mislocalization in the cell. PMID- 14550266 TI - Localization of HSP70, Cdc2, and cyclin B in sea urchin oocytes in non-stressed conditions. AB - In Paracentrotus lividus embryos, a Mediterranean sea urchin species, HSP70 is present in all the cells. During cell division it localizes under normal growth conditions on the centrosomes and on the whole isolated mitotic apparatus. Now, in situ hybridization, Western blot analyses, and immunohistochemistry show that the HSP70 mRNA is present in both small and large P. lividus oocytes, that all four isoforms of HSP70 can be found also in the oocytes, and that a certain amount of HSP70 localizes on asters and spindles during polar body formation. Moreover, two representative cell-cycle related proteins, cyclin B, and Cdc2, are present both in small and large oocytes, concentrating in the germinal vesicles before its breaking down. Cdc2 has been found in the cytoplasm of small oocytes and in the germinal vesicles of the large ones and then together with HSP70 on the mitotic apparatus of the dividing oocytes. PMID- 14550265 TI - Hepatitis B surface antigen immunopurification using a plant-derived specific antibody produced in large scale. AB - This paper provides an evaluation of a plant-derived HBsAg-specific antibody in the immunopurification of the recombinant HBsAg for vaccine purposes. This plant derived antibody was obtained from different batches of 100-200kg of tobacco leaves and coupled to Sepharose CL-4B with high efficiency. The plant-derived antibody immunoaffinity matrix purification behavior (elution capacity, antigen purity, purification cycles, and ligand leakage) was comparable to that of its mouse-derived monoclonal antibody homolog. This result supports the feasibility of using this plant-derived antibody for the immunopurification of the Hepatitis B surface antigen for human use, opening a new possibility to overcome the constrain of monoclonal antibody production in mice. PMID- 14550267 TI - A novel RNA splicing-mediated gene silencing mechanism potential for genome evolution. AB - Over 90% of the human genome consists of non-protein-coding regions. Introns constitute most of the non-coding regions located in precursor messenger RNAs (pre-mRNAs). During pre-mRNA maturation, the introns are excised out of mRNA and thought to be completely digested prior to translation. If the introns were merely metabolic "leavings," why would the genome hold such a large amount of extraneous genetic materials? Here we show a novel posttranscriptional gene silencing system identified within mammalian introns. By packaging human spliceosome-recognition sites along with an exonic insert into an artificial intron, we observed that the splicing and processing of such an exon-containing intron in either sense or antisense conformation produced equivalent gene silencing effects, while a palindromic hairpin insert containing both sense and antisense strands resulted in synergistic effects. These findings may explain how cells respond to the presence of transgenic introns that are homologous to pre existing exons during genomic evolution. PMID- 14550268 TI - Splice variant in TCRzeta links T cell receptor signaling to a G-protein-related signaling pathway. AB - The T cell receptor zeta chain is required for efficient receptor expression and contributes to T cell receptor-mediated activation of ZAP-70 and PLC-gamma1 as well as other signaling functions. A splice variant of zeta has been described which contains a 3bp insert coding for a glutamine in the cytoplasmic domain. The variant, here designated zeta-Q, is abundant, comprising 20-50% of zeta transcripts in humans, and production of the two isoforms is conserved among distantly related vertebrate species. Analysis of the peptide region in which the insert occurs reveals an unexpected homology with G-protein gamma chains. Transfection studies suggest that disruption in the alignment of three conserved prolines by the insertion of an extra glutamine impairs TCR-mediated PLC activation. Experiments with human lymphocytes suggest that zeta-Q message undergoes upregulation following cellular activation. Our data suggest that regulation of the relative levels of these two transcripts is related to an ancient mechanism which functions to raise the number of receptors required to produce cellular activation during the course of prolonged cellular stimulation, perhaps through a G-protein-related pathway. PMID- 14550269 TI - The ITIM-bearing CLECSF6 (DCIR) is down-modulated in neutrophils by neutrophil activating agents. AB - The recently discovered CLECSF6 protein displays the features of a receptor involved in the down-modulation of leukocyte activation. Although CLECSF6 has been the focus of the interest of many researchers lately, a Western blot characterization of the protein is still lacking. This highly reduces our ability to gain full knowledge of the biological relevance of this protein in cell responses. We produced two rabbit polyclonal antisera that detected a glycosylated protein at approximately 35kDa in neutrophils. Four different CLECSF6 mRNA species have been discovered to date. When deglycosylated, the protein displayed the molecular weight expected for the longest CLECSF6 form. Neutrophil membrane fractions were strongly enriched in the protein. We showed a down-modulation of the expression of this protein in neutrophils treated with granulocyte-macrophage-colony stimulating factor (GM-CSF), tumor necrosis factor (TNF-alpha), lipopolysaccharide (LPS), and interleukin (IL)-4. This work supports the hypothesis that CLECSF6 is involved in the control of inflammation in neutrophils. PMID- 14550270 TI - Regulation of osteoclast protease expression by RANKL. AB - Receptor activator of NF-kappaB ligand (RANKL) is essential for osteoclast (OC) differentiation/activation and functions through its receptor RANK at the surface of the osteoclastic cells. This study investigated for the first time the direct effects of hRANKL on protease/protease inhibitor expressions and protease activities in purified rabbit osteoclast cultures, using semi-quantitative RT PCR, gelatin zymography, and enzymatic assays. RANKL was shown to exert in vitro pro-resorptive effects by increasing osteoclast marker expressions (Tartrate resistant acid phosphatase (TRAP) and cathepsin K), MMP-9 expression, and pro-MMP 9 activity and by diminishing TIMP-1 expression, leading to an up-regulation of the MMP-9/TIMP-1 ratio. PMID- 14550272 TI - Binding characteristics of SSR180575, a potent and selective peripheral benzodiazepine ligand. AB - The peripheral benzodiazepine receptor (PBR), has been recently shown to play a key role in the regulation of the mitochondrial process leading to apoptosis, which occurs during cardiac ischemia. The present work shows that SSR180575, a novel PBR ligand of potential interest in pathological cardiovascular indications, irreversibly and specifically binds with high affinity on both rat heart mitochondria and on a cell line transfected with the human PBR (K(d)=1.95+/ 0.22 and 4.58+/-0.83nM, respectively). In conclusion, SSR180575 is a specific and potent PBR ligand which irreversible binding to PBR appears of high interest in various therapeutic indications where apoptosis occurs. PMID- 14550271 TI - PGC-1-related coactivator and targets are upregulated in thyroid oncocytoma. AB - Thyroid oncocytomas are tumors characterized by dense mitochondrial accumulation, the cause of which is currently unknown. Members of the PGC-1 coactivator family have been identified as important mediators of mitochondrial biogenesis because of their ability to activate nuclear genes encoding mitochondrial proteins. We have investigated the influence of the PGC-1 related coactivator (PRC) on the high mitochondrial content observed in oncocytoma by quantifying the transcripts of PRC, the nuclear respiratory factor 1 (NRF-1) and the mitochondrial transcription factor A (TFAM), in 30 oncocytic tumors and corresponding normal tissues. The three genes studied were found to be significantly overexpressed in thyroid oncocytomas, concomitantly with an increase in cytochrome oxidase activity and mitochondrial DNA (mtDNA) content. However, no mtDNA variant in the D-loop region appeared to be involved in oncocytic development. We conclude that overexpression of the PRC pathway is responsible for mitochondrial proliferation in the context of thyroid oncocytoma. PMID- 14550273 TI - Analysis of T-cell receptor Vbeta gene from infiltrating T cells in insulitis and myocarditis in encephalomyocarditis virus-infected BALB/C mice. AB - Encephalomyocarditis (EMC) virus induces insulin-dependent diabetes and myocarditis in several strains of mice. The T-cell receptor (TCR) Vbeta genes of infiltrating T cells in the pancreas and myocardium of BALB/C mice infected with EMC virus D-variant (EMC-D virus) were analyzed. Using a nested two-step polymerase chain reaction (PCR), TCR Vbeta cDNAs were cloned and sequenced. Two and four kinds of TCR Vbeta clones were obtained from T cells infiltrating into the pancreas and myocardium of BALB/C mice infected with EMC-D virus, respectively. The infiltrating lymphocytes in the diabetic mice expressed Vbeta 8.1, 8.2, and 8.3 genes predominantly. Previously, the use of Vbeta 8.2 has been reported in autoimmune diseases such as murine experimental allergic encephalomyelitis (EAE) and non-obese diabetic (NOD) mouse. This study suggests that mice infected with EMC virus are a useful animal model for autoimmune diseases such as insulin-dependent diabetes. PMID- 14550274 TI - Integrin-linked kinase is a positive mediator of L6 myoblast differentiation. AB - Overexpression of ILK in L6 myoblasts results in increased ILK kinase activity, stimulating myotube formation and induction of biochemical differentiation markers. Expression of a dominant negative ILK mutant, ILK(E359K), inhibits endogenous ILK activation and L6 differentiation. Cell cycle analysis of ILK(E359K) cells cultured in serum-free conditions indicates significant apoptosis (11-19% sub-diploid peak) which is not seen in insulin treated cells. Expression of ILK variants does not have significant effects on S-phase transit, however. Known targets of ILK, PKB/Akt or glycogen synthase kinase 3beta are not obviously involved in ILK-induced L6 differentiation. Insulin-stimulated phosphorylation of PKB at Ser473 is unimpaired in the ILK(E359K) cells, suggesting that PKB is not a myogenic target of ILK. Inhibition of GSK3beta by LiCl blocks L6 myogenesis, indicating that ILK-mediated inhibition of GSK3beta is not sufficient for differentiation. Our data do suggest that a LiCl-sensitive interaction of ILK is important in L6 myoblast differentiation. PMID- 14550275 TI - Angiostatin antagonizes the action of VEGF-A in human endothelial cells via two distinct pathways. AB - Angiostatin consisting of the first four-kringle domains of the plasminogen potently inhibits angiogenesis in vitro and in vivo. However, the molecular mechanism of action whereby angiostatin mediates its inhibitory effect on proliferating endothelial cells remains elusive. We therefore used the proliferating cultured human umbilical vein endothelial cells (HUVECs) promoted by vascular endothelial growth factor A to identify the endogenous signaling elements that mediate the antiangiogenic effect of angiostatin. Treatment of HUVEC with angiostatin at a concentration known to inhibit cell proliferation and induce apoptosis resulted in induction of p53-, Bax-, and tBid-mediated release of cytochrome c into the cytosol. In addition, angiostatin also activated the Fas mediated apoptotic pathway in part via up-regulation of FasL mRNA, down regulation of c-Flip, and activation of caspase 3. These results suggest that the anti-angiogenic action of angiostatin is likely mediated by two distinct signaling pathways, one intrinsic mediated by p53 while the other extrinsic involved in FasL engagement and mitochondria dysfunction. PMID- 14550276 TI - Selection of low-molecular-mass trypsin and chymotrypsin inhibitors based on the binding loop of CMTI-III using combinatorial chemistry methods. AB - Using a combinatorial chemistry approach, a decapaptide library containing the N terminal fragment of trypsin inhibitor CMTI-III was synthesized by the solid phase method. The peptide library was screened for trypsin and chymotrypsin inhibitory activity applying the iterative method in solution. Two decapeptides were selected and resynthesized for each enzyme. The association equilibrium constants ((1.1+/-0.2)x10(8) and (7.3+/-1.6)x10(7)) determined for peptides with trypsin inhibitory activity indicate that they are 3-4-fold less active than the CMTI inhibitors. On the other hand, they are significantly more effective as compared with the starting sequence. Two peptides selected as chymotrypsin inhibitors displayed about 10 times higher activity (1.7+/-0.4)x10(7) and (1.1+/ 0.2)x10(7), respectively) than those monosubstituted in position P(1) of the CMTI III analogue. Considering low molecular weight of peptides selected and the lack of conformational constraints in their structures, the results are promising. They are good templates as starting sequences for further selection of small, peptidomimetic proteinase inhibitors. PMID- 14550277 TI - A novel non-containing-nitrogen bisphosphonate inhibits both in vitro and in vivo angiogenesis. AB - Bisphosphonates (BP) are powerful inhibitors of bone resorption and are widely used in the treatment of patients with metastasis-induced osteolysis. In the present study, we show that a novel non-nitrogen-containing BP (BP7033) that exhibits antitumor activity is a potent inhibitor of both in vivo and in vitro angiogenesis. When administered to mice, BP7033 inhibited tumoral angiogenesis (65% at 0.06mg/injection) as well as tumor growth (65% at 0.006mg/injection) in a tumor model of A431 cells xenografted in nude mice, with no sign of toxicity. Additionally, in vivo angiogenesis induced by vascular endothelial growth factor containing Matrigel implants was reduced by 90% in the presence of BP7033 (0.6mg/plug). In vitro, BP7033 inhibited proliferation of human umbilical vein endothelial cells (HUVEC) (IC(50) value 3x10(-4) M) and completely prevented the formation of capillary-like tubules by HUVEC in Matrigel. Moreover, treatment of A431 cells by BP7033 induced an inhibition of Ras processing and a decrease in the secretion of both vascular endothelial growth factor and matrix metalloproteinase-2, two well-known stimulators of the proliferation and migration of endothelial cells. These findings indicate that this new BP compound has marked antiangiogenic properties and thus represents a promising candidate for treatment of malignant diseases with an angiogenic component. PMID- 14550278 TI - Transcription factor Nrf2 is required for the constitutive and inducible expression of multidrug resistance-associated protein 1 in mouse embryo fibroblasts. AB - The nuclear factor-E2 p45-related factor (Nrf) 2 is a transcription factor for the antioxidant responsive element-mediated induction of enzymes responsible for conjugation. Since multidrug resistance-associated protein1 (Mrp1/Abcc1) is an ATP-binding cassette transporter which plays an important role in the cellular extrusion of conjugated metabolites and, therefore, acts synergistically with conjugating enzymes for the cellular detoxification of xenobiotics, we examined the possibility that Nrf2 is also involved in the expression of Mrp1 in mouse embryo fibroblasts. The constitutive expression levels of Mrp1 mRNA and protein were significantly lower in Nrf2 (-/-) cells compared with those in wild type cells. In addition, significant induction by diethyl maleate was observed in wild type, but not in Nrf2 (-/-), cells, suggesting the involvement of Nrf2 in both the constitutive and inducible mRNA and protein expression of Mrp1. In addition, the uptake of [3H]2,4-dinitrophenyl-S-glutathione, a typical substrate of Mrp1, into isolated membrane vesicles also demonstrated that Nrf2 regulates the transport activity of glutathione conjugates in mouse fibroblasts. This is the first demonstration that Nrf2 is required for the constitutive and inducible expression of Mrp family proteins. PMID- 14550279 TI - Ghrelin improves endothelial dysfunction through growth hormone-independent mechanisms in rats. AB - Ghrelin is a novel growth hormone (GH)-releasing peptide which was isolated from the stomach. We have reported that ghrelin causes vasorelaxation in rats through GH-independent mechanisms. We investigated whether ghrelin improves endothelial dysfunction. Ghrelin was subcutaneously administered to GH-deficient rats for three weeks. After isolation of the thoracic aorta, aortic ring tension was measured to evaluate vasorelaxation. Acetylcholine-induced vasorelaxation was impaired in GH-deficient rats given placebo compared to that in normal rats given placebo. GH-deficient rats treated with ghrelin, however, showed a significant increase in the maximal relaxation as compared with those given placebo. This improvement by ghrelin was inhibited by N(G)-nitro-L-arginine methyl ester, a nonselective nitric oxide synthase (NOS) inhibitor. Western blot analysis demonstrated that treatment with ghrelin increased endothelial NOS (eNOS) expression in the aorta of GH-deficient rats. These results suggest that administration of ghrelin improves endothelial dysfunction and increases eNOS expression in rats through GH-independent mechanisms. PMID- 14550280 TI - Common genetic variation in a basal promoter element alters DDAH2 expression in endothelial cells. AB - Synthesis of the vasodilator nitric oxide (NO) can be inhibited by the endogenous methylarginines L-NMMA and ADMA. ADMA is elevated in a number of cardiovascular disorders in which NO availability is reduced. Elimination of ADMA from the body occurs primarily by enzymatic breakdown through the action of DDAH, of which two isoforms exist, DDAH1 and DDAH2. In this study we have identified a core promoter region of the DDAH2 gene, and transcription factor sites that play an important role in the regulation of DDAH2 expression. Using PCR-SSCP analysis we also identified six common polymorphisms. One of these polymorphisms (an insertion/deletion at position -871) within the core promoter element influenced basal transcription. The discovery of a functional polymorphism within the DDAH2 promoter suggests that there may be common, individual differences in the ability to metabolise ADMA in vivo, that in turn, might underlie susceptibility to cardiovascular disease. PMID- 14550281 TI - Aberrant methylation of the specific CpG island portion regulates cyclooxygenase 2 gene expression in human gastric carcinomas. AB - Although it has been well established that overexpression of cyclooxygenase-2 (Cox-2) favors tumorigenesis and metastasis, the molecular mechanism that regulates Cox-2 expression has not been well defined in gastric carcinoma. Aberrant methylation of the CpG island is known to be one of the powerful mechanisms for the suppression of gene expression, and usually, CpG islands are very rich in promoter region and exon-1 region. But, it is controversial whether Cox-2 gene expression is regulated by methylation of promoter region or exon-1 region. In this study, we examined whether the hyper-methylation mediated transcriptional silencing of Cox-2 also occurred in human gastric carcinoma tissues and which portion of CpG island methylation is important in Cox-2 gene expression. Genomic DNAs from human gastric carcinoma tissues were treated with three methylation-sensitive restriction enzymes and then Southern blot analysis was performed. Out of 30 primary gastric tumor samples, 26 cases (86.6%) showed overexpression of Cox-2. Four cases (13.3%) with relatively decreased Cox-2 gene expression were associated with the presence of aberrant methylation of Cox-2 CpG island. We also found that methylation of promoter region and not exon-1 region is related with the transcriptional silencing of Cox-2 in gastric carcinoma cancer by detailed methylation mapping using bisulfite sequencing analysis. Our results suggest that the DNA methylation-mediated transcriptional silencing of Cox-2 is a predominant mechanism for the down-regulation of Cox-2 expression in human gastric carcinoma. Furthermore, the results suggest that methylation of not exon-1 region but promoter region is important to regulation of Cox-2 gene expression. PMID- 14550282 TI - [LysB3, GluB29] insulin: a novel insulin analog with enhanced beta-cell protective action. AB - Insulin receptor substrate (IRS)-2 has been implicated in the promotion of beta cell survival. Here we tested the hypothesis that the novel analog [LysB3, GluB29] insulin (insulin glulisine, IG) might mediate an enhanced beta-cell protective effect due to its unique property of preferential IRS-2 phosphorylation. We assessed IRS activation by IG and its anti-apoptotic activity against cytokines or palmitic acid in comparison to insulin, insulin analogs, and insulin-like growth factor (IGF)-I using INS-1 cells. IG induced a prominent IRS 2 activation without significant IRS-1 stimulation. The marked cytokine- and fatty acid-induced apoptosis was strongly (55-60%) inhibited by IG both at the level of caspase 3 activation and nucleosomal release, with only 15% inhibition of apoptosis by regular insulin. At 1nM, insulin, insulin aspart, and insulin lispro were much less effective compared to IG. In conclusion, the prominent anti apoptotic activity of insulin glulisine might serve to counteract autoimmune- and lipotoxicity-induced beta-cell destruction. PMID- 14550284 TI - Covalent modification of cysteine 193 impairs ATPase function of nucleotide binding domain of a Candida drug efflux pump. AB - N-ethylmaleimide (NEM) impairs the ATPase function of N-terminal NBD of Candida drug resistance gene product Cdr1p. To identify the reactive cysteine(s) for such a contribution, we adopted a three-arm approach that included covalent modification, cysteine mutagenesis, and structure homology modeling. The covalent modification results clearly indicate the ability of NEM and iodoacetic acid (IAA) to potently inhibit the ATPase activity of N-terminal NBD. Since this domain contains five cysteine residues in its sequence, we mutated each and found four of these (C325A, C363A, C402A, and C462A) to stay sensitive to NEM/IAA modification and influence ATPase activity, while C193A mutation completely abrogated the catalytic function. The structural homology modeling data further validate these biochemical findings by ruling out any plausible interactions within the cysteine residues, and deriving the importance of Cys-193 in lying at a bond length clearly feasible to interact with ATP and divalent cation to critically influence ATP hydrolysis. PMID- 14550283 TI - Identification of urotensin II-related peptide as the urotensin II-immunoreactive molecule in the rat brain. AB - Urotensin II (UII) has been reported as the most potent known vasoconstrictor. While rat and mouse orthologs of UII precursor protein have been reported, only the tentative structures of UII peptides of these animals have been demonstrated, since prepro-UII proteins lack typical processing sites for their mature peptides. In the present study, we isolated a novel peptide, UII-related peptide (URP), from the extract of the rat brain as the sole immunoreactive substance to anti-UII antibody; the amino acid sequence of the peptide was determined as ACFWKYCV. cDNAs encoding rat, mouse, and human precursor proteins for URP were cloned and revealed that the sequences of mouse and human URP peptides are the same as that for rat URP. Prepro-URP gene is expressed in several rat tissues such as those of the thymus, spleen, testis, and spinal cord, although with lower levels than the prepro-UII gene. In the human, the prepro-URP gene is expressed comparably to prepro-UII in several tissues except the spinal cord. URP was found to bind and activate the human or rat UII receptors (GPR14) and showed a hypotensive effect when administered to anesthetized rats. These results suggest that URP is the endogenous and functional ligand for UII receptor in the rat and mouse, and possibly in the human. We also describe the preparation of specific monoclonal antibodies raised against UII peptide and the establishment of a highly sensitive enzyme immunoassay system for UII peptides. PMID- 14550285 TI - Vitamin D3-dependent pathway regulates TACO gene transcription. AB - Recently, gene coding for tryptophan-aspartate containing coat protein (TACO) has been recognized to play a crucial role in the survival of Mycobacterium tuberculosis within human macrophages. Since regulation of TACO gene is still poorly understood, the present study was addressed to explore the role of vitamins (A, C, D, and E) in TACO gene transcription. Such a study revealed that synergistic action of vitamin D(3) and retinoic acid (RA) had inherent ability to down-regulate TACO gene transcription in human macrophages. Based upon these results, we propose that synergistic action of RA+vitamin D may be of importance in the prevention/control of M. tuberculosis infection. PMID- 14550286 TI - Stimulation of PAI-1 and adipokines by glucose in human adipose tissue in vitro. AB - Adipokines such as Plasminogen activator inhibitor-1 (PAI-1), interleukin (IL)-8, and tumor necrosis factor (TNF)-alpha are elevated in patients with obesity, insulin resistance, and type 2 diabetes. In the present study, we investigated whether glucose affected the production of these adipokines in human adipose tissue in vitro. Glucose (up to 35mM) increased secretion of PAI-1 (p<0.01) and IL-8 (p<0.01), but not TNF-alpha, in a dose- and time-dependent manner. Half maximal stimulatory concentration of glucose was about 1mM. Glucosamine (5mM) decreased production of PAI-1 (p<0.05) and IL-8 (p<0.05), indicating that the hexosamine biosynthesis pathway is not involved in the glucose-induced increment in adipokine secretion. The present data demonstrate that glucose increases PAI-1 and IL-8 secretion. However, glucose concentrations above 5mM had no additional effects on adipokine secretion, suggesting that mechanisms other than diabetes/insulin resistance-related hyperglycemia may be involved in the observed elevation of these adipokines. PMID- 14550287 TI - Sphingosine 1-phosphate and cell migration: resistance to angiogenesis inhibitors. AB - Naturally occurring angiogenesis inhibitors can inhibit different steps of the angiogenic process, such as endothelial cell migration. However, the mechanisms underlying this inhibition have not been elucidated. We demonstrate that migration of human umbilical vein endothelial cells induced by the potent endothelial cell chemoattractant sphingosine 1-phosphate is refractory to inhibition by well-characterized angiogenesis inhibitors such as endostatin and plasminogen-related protein-B. Our data support the contention that for effective blockage of tumor-induced angiogenesis, antagonists of both G protein-coupled receptor signaling and receptor tyrosine kinase signaling must be combined. PMID- 14550288 TI - Carboxyterminal cleavage of the chemokines MIG and IP-10 by gelatinase B and neutrophil collagenase. AB - Proteolytic processing is an important regulatory mechanism for chemokines. Matrix metalloproteinases (MMPs), such as gelatinase A/MMP-2 and gelatinase B/MMP 9, are known to process the aminoterminal end of various chemokines, including interleukin-8 (IL-8/CXCL-8) and monocyte chemotactic protein-3 (MCP-3/CXCL-7). In the present study, two proteases, gelatinase B and neutrophil collagenase/MMP-8, are shown for the first time to process the carboxyterminal end of two chemokines, monokine induced by interferon (IFN)-gamma (MIG/CXCL-9) and IFN inducible protein-10 (IP-10/CXCL-10). Neutrophil collagenase degrades MIG into small fragments and cleaves IP-10 behind positions 71 and 73. Gelatinase B degrades IP-10 and cleaves MIG at three different sites in its extended carboxyterminal region. This results in the formation of MIG(1-94), MIG(1-93), and MIG(1-90). In general, gelatinase B was more efficient than neutrophil collagenase in processing these chemokines. Alignment of the CXC chemokines with the respective cleavage sites by both MMPs identified the ELR motif as a possible determinant for amino terminal cleavage by these MMPs. PMID- 14550289 TI - Intravascular insulin gene delivery as potential therapeutic intervention in diabetes mellitus. AB - We assessed therapeutic potential of intravascular insulin gene delivery in a diabetic murine model. The rat proinsulin-1 gene cDNA engineered to harbor furin consensus cleavage sequences was inserted into EBV-based plasmid vectors that contained CAG promoter or multimerized rat insulin promoter (RIP). Normal or streptozotocin (STZ)-induced diabetic mice were given an injection of the plasmids via the tail vein under high pressure. Transfection of the CAG proinsulin construct markedly improved hyperglycemia of diabetic mice, accompanied by a considerable increase in serum insulin concentrations. Although the RIP-plasmid failed to reduce fasting blood glucose, the glucose tolerance test and RT-PCR analysis revealed that insulin production was regulated in the liver in a blood glucose level-dependent manner. The present results suggest a potential therapeutic means of controlling DM. PMID- 14550290 TI - Amyloid beta peptide (Abeta42) activates PLC-delta1 promoter through the NF kappaB binding site. AB - The abnormal deposition of amyloid beta peptide (Abeta) is a hallmark of Alzheimer's disease (AD). Phospholipase C-delta1 (PLC-delta1) is also known to abnormally accumulate in the brains of AD patients, but no report has addressed the relationship between these two events. This study investigated the effect of Abeta42 on the PLC-delta1 expression in human neuroblastoma cell lines. The PLC delta1 mRNA level was increased by treatment with Abeta42 in a RT-PCR analysis. In the reporter assay, Abeta42 was found to activate the PLC-delta1 promoter activity in a dose-dependent manner. A novel NF-kappaB binding site in the PLC delta1 promoter appeared to be responsible for the Abeta42 activity. First, the dominant negative forms of the NF-kappaB activating molecules, dominant negative TGF-beta activated kinase 1 (dnTAK1) and dnNIK (dominant negative NF-kappaB inducing kinase), abolished the Abeta42 activity in the reporter assay. Second, the Abeta42 augmented a factor binding on the NF-kappaB site in the electrophoretic mobility shift assay (EMSA), which was abolished by a molar excess of the unlabeled consensus NF-kappaB oligonucleotide. These results suggest that the PLC-delta1 promoter is under the control of NF-kappaB, which mediates the expression of PLC-delta due to the Abeta42 treatment. PMID- 14550291 TI - Rapid response of cardiac obscurin gene cluster to aortic stenosis: differential activation of Rho-GEF and MLCK and involvement in hypertrophic growth. AB - Obscurin and obscurin myosin light chain kinase (MLCK) are two recently identified muscle proteins encoded by the same gene cluster. The production of obscurin, which contains a Rho-guanine exchange factor (GEF)-like sequence, and obscurin-MLCK by this cluster suggests that these novel genes may be involved in signal transduction cascades that control adaptive and compensatory responses of the heart. The goal of the present study was to investigate the transcriptional response of the obscurin gene cluster to the initiation of myocardial hypertrophy induced in mice by aortic constriction. The transcriptional activity of the obscurin genes was examined using reverse-transcriptase primed quantitative PCR. We found that the transcripts encoding the obscurin Rho-GEF and the obscurin-MLCK internal serine-threonine kinase II (SK II) domains were significantly upregulated following aortic constriction. The expression of Rho-GEF-containing transcripts at different stages of the hypertrophic growth exceeded the control levels by 2- to 6-fold. Following the induction of hypertrophy, the quantity of the SK II-encoding transcripts increased 10-fold by 24h and 16-fold by 48h, then decreased by day 7, and returned to the control level by day 56. The quantity of the carboxy terminal obscurin-MLCK transcripts encoding for SK I increased 2-fold by day 2 and returned to the control values at later stages. Immunolocalization of obscurin, which contains Rho-GEF domain, in cardiomyocytes during pharmacologically induced hypertrophic growth in vitro demonstrated that the expression was topographically associated with the growing myofibrils and with the sites of initiation and progression of myofibrillogenesis at the periphery of the sarcoplasm. This suggests that upregulation of obscurin synthesis is associated with the formation of additional amounts of contractile structures during cardiac hypertrophy. Thus, the obscurin gene cluster represents a new example of an operon that encodes differentially regulated structural and signaling proteins implicated in the control of assembly and adaptive remodeling of myofibrils during normal and hypertrophic growth. PMID- 14550292 TI - Independent signals determine the subcellular localization of NEP in prostate cancer cells. AB - NEP (Neutral endopeptidase 24.11) is a cell surface enzyme that hydrolyzes bioactive neuropeptides implicated in the transition from androgen-dependent prostate cancer (PC) to androgen-independent PC. We report the cloning and sequence analyses of NEP cDNAs from human androgen-responsive LNCaP PC cells and prostatic stromal cells. To investigate the functional role of a nuclear localization sequence (NLS) detected within the N-terminus and of an endoplasmic reticulum retention signal within the C-terminus, NEP-GFP expression vectors were constructed containing the whole NEP gene, fragments encoding the N-terminus/C terminus of the protein (5(')NEP-GFP/3(')NEP-GFP), and 5(')NEP-GFP constructs lacking the NLS. 3(')NEP-GFP transfected cells showed plasma membrane/cytoplasmic fluorescence whereas the 5(')NEP-GFP fusion protein was also detected in the nucleus. The omission of the NLS resulted in no reduction in nuclear and an increase in cytoplasmic staining. The results suggest that the analyzed structural motifs determine the subcellular distribution of NEP in epithelial LNCaP PC cells and stromal prostatic cells and therefore could be responsible for the altered cellular localization of NEP observed in PC. PMID- 14550293 TI - Comparative studies of resistin expression and phylogenomics in human and mouse. AB - Resistin is a newly identified adipocytokine that has been proposed to be a link between obesity and type 2 diabetes based on animal studies. However, the role of resistin in the pathogenesis of insulin resistance associated with obesity in humans remains unclear. We comparatively and quantitatively studied the tissue distributions of resistin mRNA between human and mouse. The expression level of resistin mRNA in human adipose tissue is extremely low but detectable by real time PCR and is about 1/250 of that in the mouse. Remarkably, resistin mRNA is abundant in human primary acute leukemia cells and myeloid cell lines U937 and HL60, but not in the Raw264 mouse myeloid cell line. Resistin expression in U937 cells was not affected by lipopolysaccharide (LPS) or by ciglitazone, a PPARgamma ligand. Phylogenomics revealed that the human resistin gene is the ortholog of its murine counterpart and is located in a region of chromosome 19p13.3, which is syntenic to mouse chromosome 8A1. In addition to the resistin-like molecule (RELM) sequences already reported, bioinformatics analysis disclosed another RELM sequence in the vicinity of RELMbeta on human chromosome 3q13.1, but this sequence is unlikely to encode an expressed gene. Therefore, only two RELMs, resistin and RELMbeta, exist in humans, instead of the three RELMs, resistin, RELMalpha, and RELMbeta, that exist in mice. This finding provides a possible answer to the question of why only two RELMs have been cloned in humans and suggests that the RELM family is not well conserved in evolution and may function differently between species. Therefore, caution should be exercised in interpreting resistin as a link between obesity and insulin resistance in humans. The high expression of resistin in human leukemia cells suggests a hitherto unidentified biological function of resistin in leukocytes. PMID- 14550294 TI - MCP-1 deficiency is associated with reduced intimal hyperplasia after arterial injury. AB - Monocyte chemoattractant protein (MCP)-1 is abundant in smooth muscle cells (SMC) and macrophages of atherosclerotic plaques and in the injured arterial wall. MCP 1 and its receptor, CCR2, are important mediators of macrophage accumulation and atherosclerotic plaque progression. We have recently reported that CCR2(-/-) mice have a approximately 60% decrease in intimal hyperplasia and medial DNA synthesis in response to femoral arterial injury. We have now examined the response to femoral arterial injury in MCP-1(-/-) mice. MCP-1 deficiency was associated with a approximately 30% reduction in intimal hyperplasia at 4 weeks and was not associated with diminished medial DNA synthesis. Despite inducing tissue factor in SMC culture, MCP-1 deficiency was not associated with a decrease in neointimal tissue factor after injury. These data suggest that MCP-1 and CCR2 deficiencies have distinct effects on arterial injury. The effects of MCP-1 on intimal hyperplasia may be mediated largely through SMC migration. PMID- 14550295 TI - Improved methods for the acquisition and interpretation of NMR metabolomic data. AB - The high spectral congestion typically observed in one-dimensional (1D) 1H nuclear magnetic resonance (NMR) spectra of tissue extracts and biofluids limits the metabolic information that can be extracted. This study evaluates the application of two-dimensional J-resolved (JRES) spectroscopy for metabolomics, which can provide proton-decoupled projected 1D spectra (p-JRES). This approach is illustrated by an investigation of embryogenesis in Japanese medaka (Oryzias latipes), an established fish model for developmental toxicology. When combined with optimized spectral pre-processing,(2) including a 0.005-ppm bin width for data segmentation and a logarithmic transformation, the reduced congestion in the p-JRES spectra increases the likelihood that a specific metabolite can be accurately integrated and thus increases the extractable information content of the spectra. Principal components analysis of the p-JRES spectra reveals the concept of a developmental trajectory that summarizes the changes in the NMR visible metabolome throughout medaka embryogenesis. Advantages and potential disadvantages of the p-JRES approach are discussed. PMID- 14550296 TI - Recombinant leptin in the hypothalamic response to late fasting. AB - The aim of the current study was to gain further insight into the implication of leptin in the regulation of hypothalamic gene expression during long-term food deprivation with emphasis on phase 3 of fasting (P3, late protein breakdown). Among plasma parameters, glucose, non-esterified fatty acids, and insulin levels tended to be decreased by leptin infusion, whilst corticosterone levels remained unchanged. From Northern blot analysis, NPY, AGRP, and MCH mRNA gene expressions were differentially regulated during prolonged fasting in leptin-perfused rats. In comparison with fed animals, NPY, AGRP, and MCH mRNA levels in P3 rats treated with leptin either remained stable or increased slightly. Regarding anorexigenic peptides (CART and POMC) and prepro-OX, fasting with leptin induced only slight changes in gene expression. Similar data have been obtained in leptin-treated fasted rats at various doses within the physiological range. We conclude that leptin and particularly low levels of plasma leptin can reasonably be considered as a constituent of a signal triggering the fasting-induced enhanced drive for refeeding in P3. PMID- 14550297 TI - Bax conformational change is a crucial step for PUMA-mediated apoptosis in human leukemia. AB - The BH3-only protein, PUMA, plays an important role in p53-mediated apoptosis. The apoptotic effect of PUMA on the mitochondria was studied using a p53 negative, human leukemia K562 cell line. Overexpression of PUMA was accompanied by an increased Bax expression, Bax conformational change, and translocation to mitochondria. A PUMA-BH3 peptide can induce Bax conformational change, cytochrome c release, and reduction in the mitochondrial membrane potential (DeltaPsi(m)) in isolated K562 mitochondria and can be inhibited by Bcl-XL. The homo-dimer of Bax/Bax was also weakly shown after mitochondria were treated with PUMA-BH3 peptide but may not be lethal for PUMA-induced apoptosis in K562 cells. Our results suggest that PUMA-induced Bax conformational change and Bax translocation to mitochondria can be separate events and the conformational change in Bax is crucial for PUMA-induced mitochondrial dysfunction. PMID- 14550298 TI - Mitochondrial deoxyribonucleoside triphosphate pools in thymidine kinase 2 deficiency. AB - Deficiency of mitochondrial thymidine kinase (TK2) is associated with mitochondrial DNA (mtDNA) depletion and manifests by severe skeletal myopathy in infancy. In order to elucidate the pathophysiology of this condition, mitochondrial deoxyribonucleoside triphosphate (dNTP) pools were determined in patients' fibroblasts. Despite normal mtDNA content and cytochrome c oxidase (COX) activity, mitochondrial dNTP pools were imbalanced. Specifically, deoxythymidine triphosphate (dTTP) content was markedly decreased, resulting in reduced dTTP:deoxycytidine triphosphate ratio. These findings underline the importance of balanced mitochondrial dNTP pools for mtDNA synthesis and may serve as the basis for future therapeutic interventions. PMID- 14550299 TI - The neural cell adhesion molecule is associated with major components of the cytoskeleton. AB - The neural cell adhesion molecule (NCAM) is a member of the immunoglobulin superfamily. Two of the three major isoforms (NCAM 140 and NCAM 180) are transmembrane glycoproteins, which differ in their intracellular domains. The present study is concerned with the identification of novel intracellular binding partners of NCAM. We expressed and purified both cytoplasmic domains of NCAM. Using ligand affinity chromatography followed by peptide mass fingerprinting, we could identify several novel binding partners of the cytoplasmic domains of NCAM 140 and 180. We present data that alpha- and beta-tubulin as well as alpha actinin 1 are associated with both NCAM 140 and 180. In contrast, beta-actin, tropomyosin, microtubuli-associated protein MAP 1A, and rhoA-binding kinase-alpha preferentially bind to NCAM 180. Furthermore, we demonstrate that inhibition of rhoA-binding kinase-alpha stimulates neurite outgrowth independently from NCAM. PMID- 14550300 TI - Molecular and physiological characterization of an invertebrate homologue of a calcitonin-related receptor. AB - Calcitonin is a key hormone involved in the regulation of calcium metabolism in vertebrates. Using oligonucleotide primers derived from consensus sequences of vertebrate calcitonin receptors, we have cloned and characterized the first representative of an invertebrate calcitonin receptor from the bivalve mollusc Crassostrea gigas. This receptor named Cg CT-R exhibits 39% amino acid sequence identity with both human calcitonin and calcitonin gene-related precursor receptors. Cg CT-R is expressed mainly in the gills and the mantle edge as well as at lower levels in muscles, digestive gland, heart, and labial palps. Transfer of animals from seawater to brackish water resulted in a significant decrease of Cg CT-R transcript levels in the gills, thus suggesting a role for ionic balance in molluscs. PMID- 14550301 TI - Induction of endogenous antioxidants and phase 2 enzymes by alpha-lipoic acid in rat cardiac H9C2 cells: protection against oxidative injury. AB - Alpha-lipoic acid (LA) has recently been reported to exert protective effects on various forms of oxidative cardiac disorders. However, the mechanisms underlying LA-mediated cardioprotection remain to be investigated. This study was undertaken to determine whether LA treatment could increase endogenous antioxidants and phase 2 enzymes in cultured cardiomyocytes, and whether such increased cellular defenses could afford protection against oxidative cardiac cell injury. Incubation of rat cardiac H9C2 cells with low micromolar concentrations of LA resulted in a significant induction of a scope of cellular antioxidants and phase 2 enzymes in a concentration- and/or time-dependent fashion. These include catalase, reduced glutathione, glutathione reductase, glutathione S-transferase, and NAD(P)H:quinone oxidoreductase-1 (NOQ1). Induction of catalase and NOQ1 was most dramatic among the above LA-inducible antioxidants and phase 2 enzymes. To further investigate the protective effects of the LA-induced cellular defenses on oxidative cardiac cell injury, H9C2 cells were pretreated with LA (25-100 microM) for 72h and then exposed to xanthine oxidase (XO)/xanthine, a system that generates reactive oxygen species (ROS), for another 24h. We observed that LA pretreatment of H9C2 cells led to a marked protection against XO/xanthine mediated cytotoxicity, as detected by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium reduction assay. The cytoprotective effects also exhibited a LA concentration-dependent fashion. Moreover, the LA pretreatment resulted in a great inhibition of intracellular accumulation of ROS in H9C2 cells following incubation with XO/xanthine. Taken together, this study demonstrates for the first time that a number of endogenous antioxidants and phase 2 enzymes in cultured cardiomyocytes can be induced by LA at low micromolar concentrations, and that the LA-mediated elevation of cellular defenses is accompanied by a markedly increased resistance to ROS-elicited cardiac cell injury. The results of this study have important implications for the cardioprotective effects of LA. PMID- 14550302 TI - Iron toxicity protection by truncated Ras2 GTPase in yeast strain lacking frataxin. AB - Yeast strain deleted for the YFH1 gene, which encodes the orthologue of human frataxin, accumulates iron in mitochondria, constitutively activates the high affinity iron import system in the plasma membrane, and is sensitive to high iron media. We have performed a genetic screen for mutants of a yfh1 deleted strain with increased resistance to high levels of iron. One of the identified mutations caused the deletion of the hypervariable C-terminal region of Ras2p GTPase. The effect of ras2 mutation on the growth of yfh1 null strain was masked by the addition of caffeine. We found that the ras2 mutation does not alter the expression of the iron regulon nor prevent mitochondrial iron accumulation in a yfh1 mutant context. The double yfh1 ras2 mutant has increased mRNA levels of CIT2 gene and augmented catalase activity. PMID- 14550304 TI - Combination of angiopoietin-1 and vascular endothelial growth factor gene therapy enhances arteriogenesis in the ischemic myocardium. AB - We hypothesised that angiopoietin-1 (Ang-1), in conjunction with vascular endothelial growth factor (VEGF) gene therapy, can enhance arteriogenesis and angiogenesis during myocardial ischemia. Mice were given a single intramyocardial injection of saline, phVEGF-A(165) and phAng-1 or a combination thereof into the non-ischemic normal heart or into the ischemic border zone of the infarcted heart. In the normal and the ischemic myocardium, gene transfer of phVEGF-A(165) alone increased the myocardial capillary density by 16% and 36%, respectively, and phAng-1 had a similar effect. In the normal heart, the ratio of arteriolar to capillary densities increased with phVEGF-A(165) and more so in the ischemic myocardium where phAng-1 also had an effect. Furthermore, the combination of plasmids induced an up to 7.5-fold increase. Transient overexpression of VEGF A(165) boosts endogenous arteriogenesis in addition to capillary angiogenesis. Ang-1 further boosts this effect at the arteriolar level. PMID- 14550303 TI - Identification of a hexapeptide binding region in the nociceptin (ORL1) receptor by photo-affinity labelling with Ac-Arg-Bpa-Tyr-Arg-Trp-Arg-NH2. AB - The interaction of Ac-Arg-Tyr-Tyr-Arg-Trp-Arg-NH(2) (HP1), a high-affinity partial agonist of the opioid receptor like (ORL1) receptor, has been investigated using the photo-labile analogue [p-benzoyl-L-Phe (Bpa)(2)]-HP1. In recombinant CHO cells expressing the human ORL1 receptor, [Bpa(2)]-HP1 binds the receptor with high affinity (K; approximately 3nM) and is as potent as HP1 in stimulating GTPgammaS binding (50-60% of nociceptin maximal effect). UV irradiation at 365nm of the complex formed by the ORL1 receptor and radio iodinated [Bpa(2)]-HP1 results in the irreversible labelling of a glycoprotein of M(r) approximately 66kDa, as determined by SDS-PAGE. Cyanogen bromide (CNBr) and enzymatic footprints of the photo-labelled receptor and an engineered receptor mutant (L113M), containing an additional CNBR cleavage site, allowed the photoreactive region to be identified as ORL1[107-113] at the C-terminal of TM helix II. In addition the presence of a disulphide bridge between Cysl23 and Cys200 has been confirmed biochemically. PMID- 14550305 TI - Galectin-1 interacts with beta-1 subunit of integrin. AB - Galectin-1, a beta-galactoside-binding dimeric lectin, is involved in adhesion, migration, and proliferation of vascular smooth muscle cells (SMC), the key steps in the development of atherosclerosis and restenosis. Here we investigated the molecular basis of the interactions between galectin-1 and SMCs. Galectin-1 modulated SMC attachment in a dose- and beta-galactoside-dependent manner. Direct binding of galectin-1 to beta1 integrin was detected by the immune precipitation of beta1 integrin after chemical cross-linking of 125I-labelled galectin-1 to the cell surface proteins. Galectin-1 transiently increased availability of beta1 integrins on the cell surface to antibodies against beta1 integrin. Incubation of SMCs with galectin-1 transiently increased the amount of the active form of beta1 integrin and tyrosine phosphorylation of two cytoskeleton-associated proteins; one of them coincided with focal adhesion kinase (FAK). Galectin-1 is likely to affect SMC adhesion by interacting with beta1 integrin on the cell surface of SMCs and inducing outside-in signalling. PMID- 14550306 TI - Ectopically expressed PDX-1 in liver initiates endocrine and exocrine pancreas differentiation but causes dysmorphogenesis. AB - To date, the potency of pancreatic and duodenal homeobox gene 1 (PDX-1) in inducing differentiation into insulin-producing cells has been demonstrated in some cells and tissues. In order to carry out efficient screening of somatic tissues and cells that can transdifferentiate into beta-cell-like cells in response to PDX-1, we generated CAG-CAT-PDX1 transgenic mice carrying a transgene cassette composed of the chicken beta-actin gene (CAG) promoter and a floxed stuffer DNA sequence (CAT) linked to PDX-1 cDNA. When the mice were crossed with Alb-Cre mice, which express the Cre recombinase driven by the rat albumin gene promoter, PDX-1 was expressed in more than 50% of hepatocytes and cholangiocytes. The PDX-1 (+) livers expressed a variety of endocrine hormone genes such as insulin, glucagon, somatostatin, and pancreatic polypeptide. In addition, they expressed exocrine genes such as elastase-1 and chymotrypsinogen 1B. However, the mice exhibited marked jaundice due to conjugated hyperbilirubinemia, and the liver tissue displayed abnormal lobe structures and multiple cystic lesions. Thus, the in vivo ectopic expression of PDX-1 in albumin-producing cells was able to initiate but not complete the differentiation of liver cells into pancreatic cells. The conditional PDX-1 transgenic mouse system developed in this study appeared to be useful for efficient screening of PDX-1 responsive somatic tissues and cells. PMID- 14550307 TI - SU9516: biochemical analysis of cdk inhibition and crystal structure in complex with cdk2. AB - SU9516 is a 3-substituted indolinone compound with demonstrated potent and selective inhibition toward cyclin dependent kinases (cdks). Here, we describe the kinetic characterization of this inhibition with respect to cdk2, 1, and 4, along with the crystal structure in complex with cdk2. The molecule is competitive with respect to ATP for cdk2/cyclin A, with a K(i) value of 0.031 microM. Similarly, SU9516 inhibits cdk2/cyclin E and cdk1/cyclin B1 in an ATP competitive manner, although at a 2- to 8-fold reduced potency. In contrast, the compound exhibited non-competitive inhibition with respect to ATP toward cdk4/cyclin D1, with a 45-fold reduced potency. The X-ray crystal structure of SU9516 bound to cdk2 revealed interactions between the molecule and Leu83 and Glu81 of the kinase. This study should aid in the development of more potent and selective cdk inhibitors for potential therapeutic agents. PMID- 14550308 TI - Metallothionein 2A interacts with the kinase domain of PKCmu in prostate cancer. AB - Prostate cancer (PC) patients die from progression to androgen independence (AI) and chemoresistance (CR). Protein kinase Cmu (PKCmu) a novel member of the PKC family of signal transduction proteins is downregulated in AI PC. Studying PKCmu interactors in the yeast two-hybrid system identified metallothionein 2A (MT 2A) as an interactor of PKCmu kinase domain (KD) in PC, which was quantified by beta gal assay, confirmed in PC cells by immunoprecipitation, and PKCmu-MT 2A co localization in vivo by immunofluorescence studies. PKCmu domain interaction studies revealed that MT 2A interacted strongly with KD, relatively weakly with C1, and failed to interact with C2, PH or kinase mutant domains. Peptide library and in silico analysis strongly suggest that MT 2A is a novel substrate of PKCmu and our data indicate that the PKCmu-MT 2A interaction depends on PKCmu kinase activity. Because metallothioneins are associated with cell proliferation and CR, the PKCmu-MT 2A interaction may contribute to CR and/or AI in PC. PMID- 14550309 TI - Beyond the knowledge deficit: recent research into lay and expert attitudes to food risks. AB - The paper reviews psychological and social scientific research on lay attitudes to food risks. Many experts (scientists, food producers and public health advisors) regard public unease about food risks as excessive. This expert-lay discrepancy is often attributed to a 'knowledge deficit' among lay people. However, much research in psychology and sociology suggests that lay risk assessments are complex, situationally sensitive expressions of personal value systems. The paper is organised around four themes: risk perception, the communication of risk, lay handling of risk, and public trust in institutions and experts. It suggests that an interdisciplinary, contextualised and psychologically sound approach to the study of risk is needed. PMID- 14550310 TI - Geometric analysis of macronutrient intake in humans: the power of protein? AB - We introduce a state-space, geometrical approach that has previously been derived and tested using non-human animals and aim to demonstrate that it has the potential to generate testable hypotheses and provide novel insights into human nutrition and diet selection. This 'Geometric Framework' is unusual in that it focuses on the interactions between dietary constituents, and thus emphasizes the manner in which organisms regulate the balance of nutrients ingested. We report results from a pilot study, which indicate that protein ingestion is more strongly regulated than carbohydrate+fat. On the basis of these results and a brief survey of other experimental and population-level data, we hypothesize that regulation of protein intake may explain more of the modern human nutritional condition than has previously been appreciated. PMID- 14550311 TI - Use of supermarket receipts to estimate energy and fat content of food purchased by lean and overweight families. AB - The aim of this study was to compare the energy and fat content of food purchased for home consumption by households comprising mainly overweight individuals (OH), with those comprising mainly lean individuals (LH). 214 supermarket shoppers and their household were recruited from a Tesco supermarket in Leeds (UK). Households collected supermarket receipts and completed a shopping diary for 28-days, and each member of the household completed a 4-day food record. OH purchased food higher in fat (38% total energy from fat) than LH, (34.9%: p=0.001) and they purchased more energy and fat per adult equivalent, per day than LH (10.05 MJ compared to 9.15 MJ: p=0.01 and 103 g compared to 86 g:p=0.001). Households were 15% more likely to be classified as OH for each additional MJ of energy purchased per person, per day, after adjusting for number of children, household size, age, sex and social class. It was concluded that food purchasing behaviour may be linked to the prevalence of obesity in households who shop at supermarkets. PMID- 14550312 TI - Meals and snacks among elderly self-managing and disabled women. AB - AIM: The aim of this study was to describe the frequency and distribution of self managing and disabled elderly women's eating events, as well as to investigate which definition/names the women had given their different eating events and to categorise these into meals and snacks. An additional aim was to study the composition of meals and snacks, and analyse the nutritional significance of these eating events in terms of energy and macronutrients. SUBJECTS: Elderly women, both self-managing (n=139) and disabled (n=63; with Parkinson's disease, rheumatoid arthritis or stroke), aged 64-88 years, and living at home participated. METHODS: A repeated 24 h recall and an estimated food diary for three consecutive days were used. RESULTS: The eating events defined by the women that were categorised as meals contributed 74% of the total daily energy intake, while snacks contributed 22-23%. The meals that the women had defined as dinner, was the most energy dense meal. The frequency of eating events not defined by the women, was 30-34%, but contributed only 3-4% of the total daily energy intake. The disabled women had a significantly lower energy content in meals and most snacks, compared to the self-managing women. CONCLUSION: The main conclusion was that elderly women still living at home had their meals distributed during the day and that these meals were characterised by individuality and flexibility. PMID- 14550313 TI - Role of conditioned taste aversion in the development of activity anorexia. AB - Activity anorexia (AA) occurs when rats are restricted to one meal period (60-90 min) each day and have unlimited access to a running wheel the rest of the time. This AA procedure also contains the conditions necessary for conditioned taste aversion (CTA). The food eaten during the meal period is the conditioned stimulus paired with the unconditioned stimulus produced by wheel running. Thus, CTA to food may account in part for the decreased eating in AA. To test this possibility, male rats were subjected to the AA procedure. They had daily 90-min exposures to their familiar chow and spent the rest of the day in running wheels (experimental condition) or home cages (control condition). A second, concurrent experiment had the same procedure except that novel, rather than familiar, food was used. In both experiments, AA occurred; the rats allowed to wheel run ate less than those in the control condition. Several days after AA training, a two food preference test assessed whether CTA occurred. Wheel running induced CTA when food was novel but not when it was familiar. Since AA is typically studied with procedures using familiar food, the present findings indicate that CTA plays little or no role in AA. PMID- 14550314 TI - Eating with your eyes: effect of appearance on expectations of liking. AB - It was hypothesised that consumers' expectations of liking for a food would be affected by its appearance both when raw and when cooked and that the impact of these expectations on actual liking for the product after eating would vary with consumer awareness of internal body states (private body consciousness). We found that consumers' expectations of liking for the food generated by the appearance of the cooked product was related to expectation of liking from viewing the raw product. Under some conditions, consumers liked a food less after consumption if a raw product that generated low expectation of liking had been presented beforehand. There was no evidence that private body consciousness modified the consumers' susceptibility to expectation effects. It was concluded that expectations of liking for a food generated by appearance both when raw and cooked influenced final evaluation of the product during consumption. PMID- 14550315 TI - Predictors of reported consumption of low-nutrient-density foods in a 24-h recall by 8-16 year old US children and adolescents. AB - The purpose of this study was to develop an explanatory model to predict the number of low-nutrient-density (LND) foods reported in a 24-h recall by US children and adolescents using data from the third National Health and Nutrition Examination Survey. The reported number of LND foods was estimated from 24-h dietary recall data for 8-16 year old respondents (n=4137; 2024 males and 2113 females). The LND foods included--baked and dairy desserts, sweeteners, salty snacks, visible/discretionary fat, and miscellaneous. The predictive ability of socio-demographic, family, weight/dieting related, life-style or food consumption related subject characteristics was determined using multiple linear regression analyses. The strongest independent negative predictor of the reported number of LND foods was the amount of nutrient-dense foods from the five major food groups. In addition, number of eating occasions reported was a significant independent positive predictor, and the weekly frequency of consuming a complete school lunch was a significant independent negative predictor of the reported number of LND foods. These models explained approximately 55% of the variance in LND food reporting in both males and females. Socio-demographic, family, body weight, or lifestyle characteristics contributed little to predicting the number of LND foods reported in a 24-h recall. PMID- 14550316 TI - Commensal eating patterns: a community study. AB - Commensality is eating with other people, and commensal eating patterns reflect the social relationships of individuals. This study examined usual meal partners in commensal units and frequency of eating with others in commensal circles among 663 adults responding to a mailed questionnaire in one community. Meal partner data revealed that most respondents ate alone at breakfast, alone or with co workers at lunch, and with family members at dinner. Commensal frequency data revealed some eating at the homes of other family members, little eating at friends' homes, and almost no eating at neighbors' homes. Few demographic variations existed in commensal eating, except that unmarried individuals more often ate breakfast and dinner alone and more often ate with friends. These finding suggest that contemporary work-oriented society may lead people to eat alone during the day but share evening meals with family, and that people maintain commensal relationships primarily with family members rather than friends or neighbors. Peoples' social worlds appear to be focused on the nuclear family, and family members are also the people they usually eat with. PMID- 14550317 TI - Prepulse inhibition of the human startle eye blink response by visual food cues. AB - We present a new paradigm to investigate cognitive processing of food cues. The study assessed if pictures of food induce prepulse inhibition (PPI) of startle eye blink, and if any such effect is sensitive to food deprivation. In a balanced cross-over design, 16 healthy male volunteers (mean age: 29 years) were tested on 2 days 1 week apart, either after a period of normal food intake (NFI) or after a 24 h period of food restriction (RFI). On each experimental day, 80 control and 20 food pictures (slides) were displayed. Noise stimuli were presented 250 ms after slide onset. Startle responses were assessed by EMG recordings of the right M. orbicularis oculi. Startle elicited during dark inter-slide periods served as control responses to calculate PPI effects. The arousing content of each picture was rated by all subjects at the end of the session. The perceived arousal effect of control slides was strongly related to their PPI effect; no impact of food access status on this association was detectable. After NFI, food slides significantly induced PPI of startle (mean: -14.5%). After RFI, arousal ratings for food pictures increased but PPI did not. These results are evidence for a pre attentive mechanism operative in the processing of visual food cues. PMID- 14550318 TI - Effects of the form of glasses on the perception of wine flavors: a study in untrained subjects. AB - Many wine connoisseur claim that the glass shape has a direct impact on wine aroma. This hypothesis was investigated in healthy volunteers (85 m, 96 f; age 19 73 years); 89 subjects received red wine, 92 subjects white wine. Subjects compared three glasses with a white wine and three glasses with a red. Glasses were of different shape but of the same height and of comparable opening diameter. All glasses had elegant stems; two glasses of 'tulip' and 'beaker'-like shapes were used in sessions with red and white wines. Different bulbous glasses were used in either red or white wine sessions. Subjects were blinded whether they received one or more wines. Intensity, hedonic tone, and quality of the wines were rated repeatedly before and after drinking. Ratings of wine odors from different glasses were influenced by glass shape. Importantly, this appeared not to relate to the esthetic impression the glasses made. Thus, the present data indicate that the shape of glasses seems to influence the perception of wine odors. PMID- 14550319 TI - Relationship of acceptability to consumption in a meal-testing environment, and the use of intake to predict product acceptability in a meal. PMID- 14550320 TI - Relationship between parental report of food neophobia and everyday food consumption in 2-6-year-old children. PMID- 14550321 TI - Consumer perceptions of genetically modified and organic foods. What kind of knowledge matters? PMID- 14550322 TI - Cephalic phase salivary response differences characterize level of food neophobia. PMID- 14550323 TI - Changes in cerebrospinal fluid signalling substances and appetite scores following 48 h fast in healthy volunteers. PMID- 14550324 TI - Time to eat: the relationship between the number of people eating and meal duration in three lunch settings. AB - We conducted an observational study of customers in three different types of lunch settings: a worksite cafeteria, a fast-food restaurant, and a moderately priced restaurant, and assessed the relationship between meal duration and the number of people eating at each table (group size). Results suggest a significant positive correlation between group size and meal duration, collapsing over eating settings. Analysis of variance yielded significant main effects of both eating setting and of group size, indicating that meal durations were longest in the moderately priced restaurant and shortest in the fast-food restaurant. An interaction between group size and eating setting indicates that the magnitude of the group size effect on meal duration differed in the different situations, with the effect of group size on duration being smallest, but still significant, in the fast-food setting compared with the other two settings. PMID- 14550325 TI - Concerns about research and prevention strategies in Munchausen Syndrome by Proxy (MSBP) abuse. PMID- 14550326 TI - Fatal constriction of an 8-year-old child by her parents' pet python: a call for amendment to existing laws on the ownership of exotic wildlife to protect children from avoidable injury and death. PMID- 14550327 TI - Cumulative environmental risk in substance abusing women: early intervention, parenting stress, child abuse potential and child development. PMID- 14550328 TI - Cumulative environmental risk in substance abusing women: early intervention, parenting stress, child abuse potential and child development. AB - OBJECTIVE: To assess the relationship between cumulative environmental risks and early intervention, parenting attitudes, potential for child abuse and child development in substance abusing mothers. METHOD: We studied 161 substance abusing women, from a randomized longitudinal study of a home based early intervention, who had custody of their children through 18 months. The intervention group received weekly home visits in the first 6 months and biweekly visits from 6 to 18 months. Parenting stress and child abuse potential were assessed at 6 and 18 months postpartum. Children's mental and motor development (Bayley MDI and PDI) and language development (REEL) were assessed at 6, 12, and 18 months postpartum. Ten maternal risk factors were assessed: maternal depression, domestic violence, nondomestic violence, family size, incarceration, no significant other in home, negative life events, psychiatric problems, homelessness, and severity of drug use. Level of risk was recoded into four categories (2 or less, 3, 4, and 5 or more), which had adequate cell sizes for repeated measures analysis. DATA ANALYSIS: Repeated measures analyses were run to examine how level of risk and group (intervention or control) were related to parenting stress, child abuse potential, and children's mental, motor and language development over time. RESULTS: Parenting stress and child abuse potential were higher for women with five risks or more compared with women who had four or fewer risks; children's mental, motor, and language development were not related to level of risk. Children in the intervention group had significantly higher scores on the PDI at 6 and 18 months (107.4 vs. 103.6 and 101.1 vs. 97.2) and had marginally better scores on the MDI at 6 and 12 months (107.7 vs. 104.2 and 103.6 vs. 100.1), compared to the control group. CONCLUSION: Compared to drug-abusing women with fewer than five risks, women with five or more risks found parenting more stressful and indicated greater inclination towards abusive and neglectful behavior, placing their infants at increased risk for poor parenting, abuse and neglect. Early home-based intervention in high-risk families may be beneficial to infant development. PMID- 14550329 TI - Differences between accounts provided by witnesses and alleged victims of child sexual abuse. AB - OBJECTIVES: To determine whether child witnesses of sexual abuse were more or less informative about the alleged incidents than alleged victims when interviewed similarly. METHOD: Twenty-six alleged victims of child sexual abuse (aged 5 to 14 years; M=9.8 years) and 26 children who had witnessed but not experienced similar events were interviewed by experienced youth investigators about the alleged abuse. Children in the two groups were matched with respect to their age, relationships with the alleged perpetrator, and seriousness of the alleged offenses. All children were interviewed using the NICHD investigative interview protocol. RESULTS: Witnesses and victims provided similar amounts of information about the incidents of abuse. Interviewers used more open-ended invitations and elicited more information using open-ended prompts from witnesses than from victims, whereas they used more risky (including suggestive) prompts when interviewing victims. DISCUSSION: These results confirm that young children can be informative witnesses about events that they have either experienced or witnessed. PMID- 14550330 TI - Emotional neglect and family structure: impact on student functioning. AB - OBJECTIVE: The goal of this study was to demonstrate the relationship between retrospective reports of child neglect, family of origin functioning, and current psychological distress. It was hypothesized that experience of childhood emotional neglect would be associated with lower levels of cohesion and adaptability in participants' family of origin and greater levels of current psychological distress. Gender effects were explored. METHOD: Retrospective reports of emotional neglect in childhood were quantified by use of a parental bonding measure, specifically by assessing level of perceived parental care. Participants were assigned to one of three groups based on level of care scores: neglect by a primary male caregiver, neglect by a primary female caregiver, and no neglect. These groups were compared on three variables: current psychological functioning, retrospective report of family of origin adaptability, and retrospective report of family of origin cohesiveness. Amount of time left in the care of others was used as a covariate based on attachment theory literature. RESULTS: Data were analyzed for 69 female and 22 male undergraduate students. Participants who reported a childhood experience of emotional neglect by a primary female caregiver described greater current psychological distress than those reporting no neglect or neglect by a primary male caregiver. Participants who experienced emotional neglect by a primary female caregiver, as compared to those reporting emotional neglect by a primary male caregiver and those reporting no neglect, described lower cohesion and adaptability in their family of origin. There were no differences between male and female participants in terms of impact of emotional neglect. CONCLUSIONS: This study suggests potential caregiver gender effects, regardless of respondent gender. Perceived neglect by female, but not male caregivers, led to psychological distress in adulthood and was associated with lower cohesion and adaptability in one's family of origin. Future research should attempt to account for the participants' expectations regarding male and female caregiver roles. PMID- 14550331 TI - Group treatments for women sexually abused as children: a review of the literature and recommendations for future outcome research. AB - OBJECTIVE: Adults with a history of sexual abuse often suffer many long-term consequences. It is important that therapists be able to provide effective treatment to address the host of issues that are presented in therapy by adult survivors of childhood sexual abuse (CSA). In order to provide the best possible treatment, therapists should know which treatments are most effective. METHOD: This paper provides a critique of the outcome research of 13 studies (six uncontrolled and seven controlled) on the treatment of adults who suffered childhood sexual abuse and discusses specific methodological strategies that can enhance the quality of such research in the future. RESULTS: Although many of the studies contain methodological limitations, the results generally indicate that group treatment helps reduce symptomatology in the short-term and at follow-up. CONCLUSION: Although outcome research has not been published on specific individual or conjoint treatment approaches for adult survivors of CSA, several outcome studies have found group treatment to be effective in the recovery of female CSA survivors. PMID- 14550332 TI - Epidemiology of child homicide in Jefferson County, Alabama. AB - OBJECTIVE: The purpose of this study was to present the epidemiology of homicide among children younger than 6 years of age in Jefferson County, Alabama. This study focused on obtaining great detail on homicides and suspicious deaths occurring within a fixed population. METHODS: For purposes of this study, cases included Jefferson County deaths attributed to "homicide" or that were "undetermined" as noted in the coroner files among children younger than 6 years of age who were born and died between January 1, 1988 and December, 31, 1998. Victim and offender characteristics were obtained from the Jefferson County Coroner/Medical Examiner Office records. Environmental factors and circumstances surrounding the death were noted as well. RESULTS: The 53 study subjects were mainly female (55%), Black (69%), younger than 2 years of age (85%), had single mothers (38%), and a history of abuse (53%). Offenders were more likely to be male (64%), Black (73%), and a parent of the victim (53%). Homicides primarily resulted from an angry impulse (61%), with hands the most common weapon (61%). CONCLUSIONS: The majority of deaths in this study occurred among children younger than age 2, with a high proportion of fatalities among Black children of unmarried mothers. The offender most often knew the victim, with half of all homicides and two-thirds of all infant homicides involving a parent. More than half of the homicides resulted from an angry impulse, while the most common scenario for deaths with undetermined intent involved the caretaker finding the child unresponsive. PMID- 14550333 TI - Comorbidity and psychiatric diagnoses among women reporting child sexual abuse. PMID- 14550334 TI - Ningaloo. PMID- 14550335 TI - State of the marine environment reports--a need to evaluate their role in marine environmental protection and conservation. AB - This paper discusses the rationale behind the preparation of state of the marine environment (SOME) reports, and the need to evaluate their role in marine environmental protection and conservation. Many SOME reports exist, and are being planned or prepared, but are the intended audiences known, are the reports recognized for their many value-added benefits during preparation, and are they being used effectively when completed? It is proposed that a detailed evaluation is needed of SOME reporting and reports, covering audience(s), role(s), influence, and lessons learned or overall benefits. PMID- 14550336 TI - Ecological condition of US Mid-Atlantic estuaries, 1997-1998. AB - The Mid-Atlantic Integrated Assessment (MAIA-Estuaries) evaluated ecological conditions in US Mid-Atlantic estuaries during the summers of 1997 and 1998. Over 800 probability-based stations were monitored in four main estuarine systems- Chesapeake Bay, the Delaware Estuary, Maryland and Virginian coastal bays, and the Albemarle-Pamlico Estuarine System. Twelve smaller estuaries within the four main systems were also assessed to establish variance at the local scale. A subset of the MAIA-Estuaries data is used here to estimate the extent of eutrophication, sediment contamination, and benthic degradation in mid-Atlantic estuaries. An Environmental Report Card and Index of Environmental Integrity summarize conditions in individual estuaries, the four estuarine systems, and the entire MAIA region. Roughly 20-50% of the region showed signs of eutrophication (high nutrients, excessive production of organic matter, poor water clarity, or depleted dissolved oxygen), 30% had contaminated sediments, and 37% had degraded benthic communities. Compared with the Environmental Monitoring and Assessment Program (EMAP)-Virginian Province study in 1990-1993, larger fractions of Chesapeake Bay (17%) and Delaware River (32%) had increased metals or organics in sediments. PMID- 14550337 TI - The influence of cyanobacteria on oil polluted intertidal soils at the Saudi Arabian Gulf shores. AB - In 1991 the second Gulf War lead to the largest oil spill in human history. Over 770 km of coastline from southern Kuwait to Abu Ali Island (Saudi Arabia) were smothered with oil and tar, erasing most of the local plant and animal communities. In the following year cyanobacteria colonized most of the oil polluted shores. In the study area at the Saudi Arabian Gulf coast north of Jubail three different processes were observed that followed the extensive formation of cyanobacterial mats. The first one is desiccation, cracking, and peeling of the cyanobacterial mats, thereby removing the uppermost part of the oiled sediment. The second is the resettlement of burrowing macrofauna like crabs and benthic animals such as gastropods, which outcompete the cyanobacteria again. The third process is further extensive growth of cyanobacteria building thick laminated mats. These layers, completely seal the surface and hence produce an anaerobic milieu which inhibits oil degradation. As long as such cyanobacteria exist, they will prevent microbial oil degradation as well as any resettlement by macrofauna. PMID- 14550338 TI - Heavy metal sedimentary record in a Galician Ria (NW Spain): background values and recent contamination. AB - Two long sediment cores were sampled at the Ferrol Ria (Galicia, NW Spain) and the heavy metal (Co, Cr, Cu, Fe, Ni, Pb and Zn) concentrations measured in order to (i) state accurate background values providing baseline relationships with respect to a reference element and (ii) to investigate the recent metal contamination trends. Background values were found to agree well with the world average values for granite/schists-genisses rocks. However, Cu, Co, Pb and Zn were found to be lower than those previously reported as background values for the Galician Rias. Results emphasize the importance of using baseline relationships with respect to a normalizing element in order to reduce the scattering of data and to allow an accurate statement of background values. The distribution of metals in the cores showed an evident enrichment in the surface layers belonging to the industrial era. Normalized enrichment factors (NEF) for copper and zinc are in the order of 3-5 (certain/severe contamination) in the surface sediments, decreasing with depth. Lead contamination has decreased in the recent years from NEF of 3-7 down to a NEF of 2 (i.e. moderate contamination), probably due to the introduction of unleaded gasolines. Chromium, cobalt and nickel NEFs were always in the <1-2 range indicating null/low contamination by these metals. PMID- 14550339 TI - Chronic effects of tributyltin on the caprellid amphipod Caprella danilevskii. AB - In order to examine the biological effects by TBT exposure, experiments involving the exposure of five levels of TBT concentrations (0, 10, 100, 1000 and 10,000 ngl(-1)) were conducted on the caprellid amphipod Caprella danilevskii, over a generation after hatching. Marked delays in growth and molting during the early developmental stage and maturation were found in both 100 and 1000 ng TBTCl l(-1) concentrations in spite of the sex. All specimens died in 10,000 ng TBTCl l(-1) within 4 days after hatching. Inhibition of maturation and reproduction such as delaying in the achievement of maturity and a decrease in the number of juveniles hatched was apparent in 10 and 100 ng TBTCl l(-1) concentrations. Furthermore, brood loss, and failure in egg formation and hatching were observed as the TBT concentration became higher. No significant changes in sex ratio were seen at any of the TBT exposure levels during hatching and maturation in the present study. This phenomenon was different from a previous study in which the proportion of females was increased with an increase in TBT concentrations in the embryonic exposure experiment. This suggests that sex disturbance is induced during the embryonic stage of the species. A drastic decrease in survival rate was observed at 10 ng TBTCl l(-1) (25%) which corresponds to the mean level in coastal waters. Therefore, the present encountering ambient TBT concentration may influence populations of C. danilevskii in the coastal environment. PMID- 14550340 TI - Sublethal effects of contamination on the Mediterranean sponge Crambe crambe: metal accumulation and biological responses. AB - The effect of low levels of pollution on the growth, reproduction output, morphology and survival of adult sponges and settlers of the sponge Crambe crambe were examined. We transplanted sponges from a control area to a contaminated site and measured the main environmental variables (chemical and physical) of both sites during the study period. Except some punctual differences in particulate organic matter, silicates, nitrates, and water motion, most environmental variables in the water were similar at both sites during the study months. Mainly copper, lead and OM concentrations in the sediment, and water motion were significantly higher at the polluted site and may be implicated in the biological effects observed: decrease in the percentage of specimens with embryos, increase in shape irregularity and decrease in growth rate. Individuals naturally occurring at the polluted site and those transplanted there for four months accumulated ten times more copper than either untouched or transplant controls. Although lead concentration in sediment did not differ between sites, native specimens from the contaminated site accumulated this metal more than untouched controls. Vanadium concentration also tended to increase in the sponges living at or transplanted to the contaminated site but this difference was not significant. C. crambe is a reliable indicator of metal contamination since it accumulates copper, lead and vanadium in high amounts. At the contaminated site, sponge growth, fecundity and survival were inhibited, whereas sponge irregularity ending in sponge fission was promoted. All these effects may compromise the structure and dynamics of the sponge populations in sheltered, metal-contaminated habitats. PMID- 14550341 TI - Exposure and time dependent DNA strand breakage in hepatopancreas of green-lipped mussels (Perna viridis) exposed to Aroclor 1254, and mixtures of B[a]P and Aroclor 1254. AB - Green-lipped mussels (Perna viridis) were exposed to Aroclor 1254 (0.5, 5 and 50 microgl(-1)) and a mixture of benzo[a]pyrene (B[a]P) and Aroclor 1254 (0.3+0.5 and 3+5 microgl(-1)) for 12 days. On day 0, 1, 3, 6 and 12, the levels of DNA strand breaks in the mussel hepatopancreas were monitored using an alkaline unwinding assay. The results were compared to the findings of a previous study in which the levels of DNA strand breakage in the same species were measured following exposure to various concentrations of B[a]P (0.3, 3 and 30 microgl( 1)). The results indicated that Aroclor 1254 at a concentration 1000 mg/l (PES-61) and 2.5 mg/l (Triton X-100) to 413.6 mg/l (PES-61) for M. beryllina. Chronic first-effect concentrations (mg/l) for the six surfactants ranged from 2.3 to 465.0 (M. beryllina) and 1.0 to >1000.0 (M. bahia) based on reductions in growth and fecundity. Few generalizations could be made concerning the results due to their variability but M. bahia was generally the more sensitive species and the toxicities of the biosurfactants were intermediate to those of the synthetic surfactants. PMID- 14550344 TI - Antioxidants in the black mussel (Mytilus galloprovincialis) as an indicator of Black Sea coastal pollution. AB - The biologically active compounds, antioxidant activities, and free radical scavenging effects of black mussel dry matter (Mytilus galloprovincialis)--(BMDM) were investigated. The extract from BMDM with absolute methanol (BMDMAMet) showed the strongest inhibition of lipid peroxidation as a function of its concentration, and was comparable to the antioxidant activity of butylated hydroxyanisole, at the same concentration of 0.2 mg/ml. The extract with 50%methanol/water (BMDM50%Met) had the weakest antioxidant activity, whereas other extracts such as 1.2 M HCl in 50%methanol/water (BMDM50%Met/HCl) exhibited an average inhibition of lipid peroxidation. BMDMAMet extract showed marked activities in free radical scavenging determined by beta-carotene bleaching (beta carotene), nitric oxide (NO*) and 2,2'-azino-bis(3-ethylbenzothiazoline-6 sulfonate) radical cation (ABTS*+ methods. The antioxidant activities of BMDMAMet extract determined beta-carotene, NO* and ABTS*+ methods showed the highest correlation coefficients (R2) such as 0.998, 0.9976 and 0.6103, respectively, with the presence of total polyphenols. BMDM50%Met had lower antioxidant ability than BMDM50%Met/HCl in different scavenging methods, indicating that the major antioxidant components in these extracts must be derived from the polyphenols. Correlation between proteins and antioxidants was very low (0.0318 and 0.0433). The mussel extracts from polluted areas have shown by all using methods higher antioxidant activity than from the clean ones. The results presented in this report indicate that the antioxidant activities of mussel extracts could possibly be an additional index of pollution characterization. PMID- 14550345 TI - Application of trichloroacetic acid (TCA) to extraction of soft body for the determination of tissue Cd, Cu, Pb and Zn in the prosobranch Hydrobia ulvae (Pennant). AB - The application of trichloroacetic acid (TCA) as a shell extractant for preparation of soft body parts with reference to tissue metal concentrations (Cd, Cu, Pb, Zn) in shellfish has been evaluated on the example of the mud snail Hydrobia ulvae, a small marine prosobranch densely present in rocky and soft bottom habitats of the eastern Atlantic. A solution of 0.1 M TCA was tested on individuals treated according to two different protocols: (1) thawed after freezing ("non-dried") and (2) thawed and air-dried to a constant weight ("dried"). Two points were investigated in detail to improve the method: individual soft tissue dry weight and tissue metal concentration following a standard digestion method. In both instances, the results were compared with those from manually dissected snails. Conditions for total shell decalcification of 60 individuals (3-4 mm long) were 5.5 h in 20 ml of 0.1 M TCA.No differences in individual soft tissue weight were observed between the treatments, indicating good efficiency of the TCA extraction with respect to weight of soft body parts. In contrast, tissue metal concentrations varied among treatments. The TCA extraction of the dried animals had a good recovery for Cd, most likely due to the lower solubility of Cd vital cellular components (proteins and mineral concretions) from the dried tissue. Satisfactory recoveries of the tissue concentrations of Cu and Pb were obtained for the non-dried individuals. This might be related to the specific distribution of metals in the organism (namely in the digestive glands and gonads) and their different chemical reactivity with TCA after the tissue was dried. Limited susceptibility of Zn-bearing protein bindings to complexing with TCA also accounts for significantly lower concentrations of Zn in the mud snail's soft tissue that was extracted. The 0.1 M TCA solution is therefore recommended for extraction of the shells of Hydrobia ulvae for tissue determination of Cd, Cu and Pb; however the treatment protocol does affect metal recovery and thus a consistent procedure should be followed. The extracted metals from the soft tissues and shells of the mud snails (on the basis of both metal concentrations and contents) were ranked in order of increasing contribution of soft body parts to the total (shell+tissue): Pb70%) accumulated in the sludge, while 47-63% of Cd, Cr, Pb, Fe, Ni and Zn remain in the treated effluent. PMID- 14550352 TI - Experimental design of Fenton and photo-Fenton reactions for the treatment of cellulose bleaching effluents. AB - Multivariate experimental design was applied to the treatment of a cellulose conventional bleaching effluent in order to evaluate the use of the Fenton reagent under solar light irradiation. The effluent was characterised by the general parameters total organic carbon (TOC), chemical oxygen demand and color, and it was analysed for chlorinated low molecular weight compounds using GC-MS. The main parameters that govern the complex reactive system: Fe(II) and H(2)O(2) initial concentration, and temperature were simultaneously studied. Factorial experimental design allowed to assign the weight of each variable in the TOC removal after 15 min of reaction. Temperature had an important effect in the organic matter degradation, especially when the ratio of Fenton reagents was not properly chosen. Fenton reagent under solar irradiation proved to be highly effective for these types of wastewaters. A 90% TOC reduction was achieved in only 15 min of treatment. In addition, the GC-MS analysis showed the elimination of the chlorinated organic compounds initially detected in the studied bleaching effluents. PMID- 14550353 TI - Removal of metal ions from the complexed solutions in fixed bed using a strong acid ion exchange resin. AB - Fixed bed removal of equimolar metal ions (Co(2+), Ni(2+), Mn(2+), Sr(2+)) from aqueous solutions using a strong-acid resin was examined. The solution contained a water-soluble complexing agent including ethylenediaminetetraacetic acid, nitrilotriacetic acid, and citric acid. Experiments were performed under different solution pH and molar concentration ratios of complexing agent to the total metals. It was shown from batch studies that the equilibrium exchange of metals and the resin mainly depended on solution pH, and partly on the type of complexing agent used. A mass transfer model was proposed to describe the breakthrough curves of the resin bed, which contained two parameters (tau and k) estimated from the observed breakthrough data. The calculated breakthrough curves agreed well with the measured ones (standard deviation <6%). In fixed bed tests at low pH (=2), the type of complexing agent had little effect on the breakthrough data. For a given complexing agent, the metal with a larger overall formation constant (K(f)) showed a smaller exchange capacity. For a given metal ion, the complexing agent with a larger K(f) also revealed a smaller exchange capacity. PMID- 14550354 TI - Aniline degradation by electrocatalytic oxidation. AB - The degradation of aniline solution in alkaline medium of pH 11.0 by electrocatalytic oxidation has been studied using an electrochemical reactor containing a SnO(2)-Sb(2)O(3)-PtO anode and a Ti cathode, both of 54 cm(2) area. Hydroxyl radicals (HO(z.rad;)) are produced at the anode, being tested with the trace catcher salicylic acid and phenol by spectrophotometery and high performance liquid chromatography. Intermediates dianiline, 4-anilino phenol and azobenzol were detected by gas chromatography-mass spectrometry. The existence of HO(z.rad;) produced in the aniline degradation was assayed with scavenger tertiary butanol. The results showed that electrocatalytic oxidation is an effective process for the degradation of aniline. A general reaction pathway that accounts for aniline degradation to CO(2) involving those intermediates is proposed. PMID- 14550357 TI - Can enquiries into severe acute maternal morbidity act as a surrogate for maternal death enquiries? AB - OBJECTIVE: To assess whether severe acute maternal morbidity (SAMM, 'near misses') can be used as a surrogate of an analysis of maternal deaths to describe the pattern of severe maternal disease and avoidable factors related to it. DESIGN: Prospective, descriptive study. SETTING: A SAMM and maternal mortality audit was conducted in three clearly defined geographical areas, consisting of rural and urban settings in South Africa. POPULATION: Indigent black African pregnant women. METHOD: Cases of SAMM and maternal deaths were collected in the areas and a comparison was made of the disease profiles and avoidable factors, missed opportunities and substandard care. MAIN OUTCOME MEASURES: The proportion of primary obstetric causes and avoidable factors in women with SAMM and maternal deaths, and the mortality indices of the primary obstetric causes of death and organ system dysfunction. RESULTS: A total of 423 women with SAMM and 128 maternal deaths were collected over two years. Demographic factors were similar between the groups except that significantly more maternal deaths had not attended any antenatal care. The primary obstetric causes of SAMM and maternal death did not correlate. The four most common causes of SAMM were complications of hypertension (27.2%), postpartum haemorrhage (18.0%), antepartum haemorrhage (12.8%) and abortion (11.3%), whereas the four most common causes of maternal death were non-pregnancy related sepsis (26.6%), complications of hypertension (23.4%), pre-existing medical disease (14.1%) and abortion (10.9%). The types of avoidable factors were similar between both groups although administrative factors occurred significantly more frequently in the maternal death group as did poor problem identification and monitoring. CONCLUSION: Review of SAMM gives a different disease pattern to that obtained from maternal death audits. However, in diagnosing inadequacies in the health system, similar information was obtained. PMID- 14550356 TI - Individualising use of aspirin to prevent pre-eclampsia: a framework for clinical decision making. PMID- 14550358 TI - The burden of disease from neonatal mortality: a review of South Asia and Sub Saharan Africa. AB - OBJECTIVE: To assess the burden of neonatal mortality in two developing regions. DESIGN: Review of secondary data collated through literature review. SETTING: Community and facility based studies and national health surveys. POPULATION: Neonatal (<28 days) population of South Asia and Sub-Saharan Africa. METHODS: Data on neonatal mortality were gathered from peer-reviewed literature, reports of the Demographic and Health Surveys and websites of country-based organisations. The base year for this study is 1995. For each country, a weighted mean neonatal mortality rate was calculated and the total number of neonatal deaths estimated. Country data were summarised over each region to estimate annual regional neonatal deaths and rates. The burden of disease from neonatal mortality was determined using a summary measure of health--healthy life years. MAIN OUTCOME MEASURES: Numbers of deaths and healthy life years (HeaLYs). RESULTS: Neonatal mortality rate for South Asia ranged from 41.9 to 56.9 per 1000 live births for 1995. Sri Lanka was an exception with a neonatal mortality rate between 16.3 and 18.6 per 1000 live births. The estimated regional neonatal mortality rate for South Asia was 46.27 per 1000 live births for 1995. There was a significant lack of data from Sub-Saharan Africa, resulting in highly variable neonatal mortality rates, ranging from 13 per 1000 live births in Kenya to 108 per 1000 live births in Senegal. The mean regional neonatal mortality for Sub Saharan Africa for 1995 was estimated at 38.8 per 1000 live births. CONCLUSION: The burden of neonatal mortality in only these two regions of the developing world represents more than 2 million annual deaths. A call for greater investments in neonatal research and health programs is a logical extension to this review of evidence. PMID- 14550359 TI - Reproductive health in rural Malawi: a population-based survey. AB - OBJECTIVES: To assess pregnancy outcome, maternal mortality and health-seeking behaviour in a rural African population and to assess the effects on these of women's education, distance from a health centre and household type. DESIGN: Descriptive population-based study. SETTING: A rural community in southern Malawi. POPULATION: All women living in the catchment area of a rural health centre. METHODS: Interviews with women in 20,649 households using structured questionnaires. MAIN OUTCOME MEASURES: Pregnancy outcome, the effect of women's education, distance from a health centre and household type on pregnancy outcome, maternal morbidity and estimates of maternal and perinatal mortality. RESULTS: Educational level was lower for women than for men. A significant association was found between educational level and fertility. Women aged 45-49 reported an average of six pregnancies with four resulting in currently living children. Successful pregnancy outcome was more likely with increased education and if the woman lived closer to the health centre. Despite living an average of 5 km from the health centre, over 90% of women attended antenatal clinic with a mean of five visits. Assistance at delivery by a trained health care worker was more likely as education increased and was less likely as distance from the health centre increased. Maternal mortality was reported at 413 per 100,000 deliveries (95% CI 144-682). The perinatal mortality rate in this population was estimated at 30 per 1000. An increased perinatal mortality was noted for women who were delivered by a female relative. Perinatal mortality rates were similar for delivery by a traditional birth attendant or a trained nurse-midwife. Education and proximity to the health centre were both associated with improved outcome. CONCLUSIONS: Many women in this rural community suffer the consequences of high pregnancy loss. Maternal and perinatal mortality are high. Improved education and skilled assistance at delivery can result in improved pregnancy outcome. Proximity of any household to a health centre has an effect on outcomes. PMID- 14550360 TI - Myometrial and placental artery reactivity alone cannot explain reduced placental perfusion in pre-eclampsia and intrauterine growth restriction. AB - OBJECTIVES: (1) To investigate a possible association between myometrial and placental artery vasoreactivity and perfusion at the basal and chorionic plates, respectively. (2) To confirm that myometrial arteries from women with pre eclampsia and intrauterine growth restriction exhibit an attenuated endothelium dependent vasodilatory response. METHODS: Women with normal pregnancy, pre eclampsia and intrauterine growth restriction had a magnetic resonance scan to assess placental perfusion using a technique called intravoxel incoherent motion. At delivery, myometrial and chorionic plate placental arteries were assessed on a wire myograph. Vessels were pre-constricted with the thromboxane mimetic U46619 and dilated with incremental doses of bradykinin. RESULTS: Pre-constricted myometrial arteries from women with pre-eclampsia or intrauterine growth restriction exhibited an attenuated vasodilatory response to bradykinin, compared with normal pregnancy (P < 0.0001). Pre-constricted placental arteries exhibited a minimal vasodilatory response in all three groups of women (P = 0.10). Maximal constrictor and vasodilatory responses of myometrial arteries were not associated with the perfusing fraction at the basal plate. Maximal constrictor and vasodilatory responses of chorionic plate placental arteries were not associated with the perfusing fraction at the chorionic plate. CONCLUSION: We confirm that myometrial arteries from women with pre-eclampsia or intrauterine growth restriction exhibit an attenuated endothelium-dependent vasodilatory response. Apart from vasoreactivity of small arteries, other factors may be involved in the control of placental perfusion. PMID- 14550361 TI - Acute increase in femoral artery resistance in response to direct physical stimuli in the human fetus. AB - OBJECTIVE: To determine whether fetal response to needling resembles the fetal 'brain sparing' response seen with hypoxaemia. DESIGN: Prospective observational study. SETTING: Centre for Fetal Care, Queen Charlotte's and Chelsea Hospital, London. POPULATION: Eighty-five pregnant women undergoing invasive procedures associated with fetal prenatal diagnosis and/or management. METHODS: The femoral artery and the middle cerebral artery pulsatility index were measured by Doppler ultrasonography before and after 89 invasive procedures (fetal blood sampling, transfusion, bladder or cyst aspiration, shunt insertion and amniocentesis, between 17 and 36 weeks). Cases in which the fetal body was transgressed were compared with 'control' fetuses undergoing invasive procedures which did not directly involve needling the fetus (amniocentesis and placental cord insertion procedures). MAIN OUTCOME MEASURES: Femoral artery and middle cerebral artery pulsatility index. RESULTS: The femoral artery pulsatility index rose after transgression [median change (Delta) 0.73; 95% confidence interval (CI) 0.51 to 0.98]. In contrast, there was no significant change in femoral artery pulsatility index after non-transgression procedures (mean Delta 0.28; 95% CI -0.20 to 0.76). Analysis confirmed the fall in middle cerebral artery pulsatility index after transgression procedures (median Delta -0.19; 95% CI -0.07 to -0.35), but there was also a significant fall in middle cerebral artery pulsatility index after non transgression procedures (mean Delta -0.47; 95% CI -0.23 to -0.71). CONCLUSIONS: The human fetus mounts a peripheral haemodynamic response to invasive procedures involving transgression of the fetal body, which is consistent with the brain sparing effect. However, the change in middle cerebral artery pulsatility index in both transgression and control procedures suggests that the changes and mechanisms may be more complex than previously thought. PMID- 14550362 TI - The cost of microwave endometrial ablation under different anaesthetic and clinical settings. AB - OBJECTIVE: To compare the costs of microwave endometrial ablation under local anaesthetic and general anaesthetic in an operating theatre and to estimate the cost of performing treatment under local anaesthetic in a dedicated clinic setting. DESIGN: The costing study was undertaken alongside a randomised controlled trial comparing the acceptability of microwave endometrial ablation using local versus general anaesthetic in a theatre setting. SETTING: Department of Gynaecology, Aberdeen Royal Infirmary, Scotland. SAMPLES: One hundred and twenty-seven women undergoing microwave endometrial ablation who had been randomly allocated to general or local anaesthetic. METHODS: Health and non health service resource use was recorded prospectively. Data on resource use were combined with unit costs estimated using standardised methods to determine the cost per patient for microwave endometrial ablation under local or general anaesthetic in theatre. A model was developed to estimate the health service cost of microwave endometrial ablation under local anaesthetic in a clinic setting. MAIN OUTCOME MEASURES: Health and non-health service costs. RESULTS: There was little difference in cost when treatments were performed under local or general anaesthetic in theatre. The median health and non-health cost of microwave endometrial ablation was 440 pounds and 120 pounds, respectively, under general anaesthetic and 428 pounds and 125 pounds per women under local anaesthetic. The health service cost of microwave endometrial ablation using local anaesthetic in a clinic setting was estimated to be 432 pounds per treatment; however, this varied from 389 pounds to 491 pounds in the sensitivity analysis. CONCLUSION: There are minimal cost savings to the patient or health service from using local rather than general anaesthetic for microwave endometrial ablation in a theatre setting. Cost modelling suggests that in a clinic setting microwave endometrial ablation has a similar cost to theatre based treatment once re-admissions for treatment under general anaesthetic are considered. Sensitivity analysis indicated that these findings were sensitive to assumptions in the model. PMID- 14550363 TI - Risk factors for stress, urge or mixed urinary incontinence in Italy. AB - OBJECTIVE: To assess risk factors for different type of urinary incontinence stress, urge or mixed-and overactive bladder without urinary incontinence. DESIGN: Case-control study. SETTING: Gynaecology clinics. POPULATION: Women attending first level gynaecological centres. METHODS: Cases were 1062 women with urinary incontinence or overactive bladder aged 40 years or more (mean age 62.3 years, range 40-88) consecutively observed in first level gynaecological centres. Controls were 1143 women (mean age 58 years, range 40-86) observed in the same centre after the identification of the cases, without any symptoms related to urinary incontinence or overactive bladder. MAIN OUTCOME MEASURES: Determinants of different types of urinary incontinence and overactive bladder. RESULTS: The risk of urinary incontinence was lower in more educated patients and increased with body mass index (BMI). The number of vaginal births was associated with the risk of stress and mixed urinary incontinence, but not urge urinary incontinence and overactive bladder. In comparison with nulliparae, the odds ratios (OR) for stress and mixed urinary incontinence were 5.4 (95% confidence interval [CI] 1.9 15.0) and 1.6 (95% CI 1.0-2.6), respectively, for women reporting one or two vaginal birth and 5.1 (95% CI 1.8-14.5) and 2.2 (95% CI 1.3-3.7), respectively, for three or more. A history of operative vaginal delivery was directly associated with the risk of stress and mixed urinary incontinence and increased the risk of urge urinary incontinence and of overactive bladder, but the latter ORs were not statistically significant. When compared with women with no birth, a history of caesarean section increased the risk of stress urinary incontinence. A history of hysterectomy, recurrent urinary infection and perineal trauma increased the risk of all types of urinary incontinence. CONCLUSIONS: Vaginal births increased the risk of stress and mixed urinary incontinence, but not of urge urinary incontinence and overactive bladder. The risk of all types of urinary incontinence was increased in women with higher BMI, with a history of hysterectomy, urinary infection and perineal traumas. These factors were not related to the risk of overactive bladder. PMID- 14550364 TI - Colonisation of extragenital sites by Candida in women with recurrent vulvovaginal candidosis. AB - OBJECTIVES: To study colonisation of extragenital sites by Candida in women with a history of recurrent vulvovaginal candidosis and to determine the association of such colonisation with clothing and sexual practice. DESIGN: A prospective case-control (current genital colonisation by candida organisms vs non colonisation) study. SETTING: Outpatient clinic, University hospital, Kiev, Ukraine and Department of Obstetrics and Gynaecology, University Hospital, Lund, Sweden. SAMPLE: Women with a history of recurrent vulvovaginal candidosis and an assumed new attack of the condition. METHODS: Eighty-six women with a history of recurrent vulvovaginal candidosis and an assumed current attack of this condition underwent culture for Candida from the genital tract, the perianal skin, the rectum, the mouth and from voided urine samples, using Sabouraud and chromogenic agar. The women were interviewed about some factors that have been considered to trigger colonisation with Candida spp. MAIN OUTCOME MEASURES: Colonisation of extragenital sites of women with recurrent vulvovaginal candidosis by different species of Candida. Correlation with oral and anal sex, and with clothing. RESULTS: Candida spp. was isolated from the mouth, the rectum, the perianal skin and from voided urine in 11(13%), 24(28%), 35(41%) and 16(19%) of women, respectively. Candida was recovered from one or more of these sites in 44(51%) of the women. The rate of culture of Candida from the extragenital sites was significantly higher (P < 0.05) in women who had a positive culture from the introitus or the vagina. The recovery rates of Candida in the 46 vaginal culture positive women were 8 (17%), 24 (52%), 34 (74%) and 16 (35%) from the extragenital sites in the order mentioned. Species of Candida other than Candida albicans occurred more often at the extragenital than the genital sites. Neither oral nor anal sex was associated with colonisation of the mouth and the rectum with Candida spp. Neither tight-fitting trousers nor underwear produced from synthetic material was associated with colonisation of the perianal skin with Candida spp. CONCLUSIONS: Extragenital sites are often colonised by Candida spp. in women with a history of recurrent vulvovaginal candidosis, but significantly more often if the genital tract is also colonised. Extragenital sites are significantly more commonly colonised with species other than C. albicans. Extragenital sites may be reservoirs for recolonisation of the genital tract in women with recurrent vulvovaginal candidosis. This has implications for treatment. PMID- 14550366 TI - Recent trends in breech delivery in the Netherlands. AB - The effect of publication of the results of the Term Breech Trial in October 2000 on the mode of delivery of babies in breech presentation in the Netherlands was subject of this study. The results showed an immediate amplifying impact on top of the already existing trend of rising caesarean section rates for term singleton breech presentations. The willingness of obstetricians in the Netherlands to adopt the conclusions of the Term Breech Trial so quickly originate from scientific, legal, psychological and emotional sources. The same amplifying impact applied for all non-term breeches and breeches from multiple gestations, but not for babies in other than breech presentation. PMID- 14550365 TI - Saline contrast hysterosonography in abnormal uterine bleeding: a systematic review and meta-analysis. AB - OBJECTIVE: To assess the diagnostic accuracy of saline contrast hysterosonography in the evaluation of the uterine cavity in women complaining of abnormal uterine bleeding. DESIGN: A systematic review and meta-analysis of diagnostic studies that compared saline contrast hysterosonography to a gold standard diagnosis based on either hysteroscopy with or without histological sampling or to hysterectomy. SETTING: University Hospital. SAMPLE: Twenty-four studies (including 2278 procedures). METHODS: Electronic databases were searched for relevant studies and references were cross checked. Validity was assessed and data were extracted independently by two authors. Heterogeneity was calculated, studies were plotted in an ROC area and data were pooled. Subgroup analysis was performed according to the validity area. MAIN OUTCOME MEASURES: The success rate of saline contrast hysterosonography. The pooled sensitivity, specificity, likelihood ratio's and post-test probabilities of saline contrast hysterosonography on the prediction of uterine cavity abnormality. RESULTS: The largest population of homogeneous data were the studies with complete verification. In these studies, the pooled sensitivity and pooled specificity of saline contrast hysterosonography in uterine cavity evaluation were respectively 0.95 (95% CI 0.93 to 0.97) and 0.88 (95% CI 0.85 to 0.92), the likelihood ratios were respectively 8.23 (95% CI 6.2 to 11) and 0.06 (95% CI 0.04 to 0.09) and the post-test probabilities were respectively 0.91 (95% CI 0.89 to 0.94) and 0.07 (95% CI 0.04 to 0.10). The overall success rate of saline contrast hysterosonography was 93% (95% CI 92% to 94%). The feasibility of saline contrast hysterosonography in postmenopausal women (success rate 86.5%, 95% CI 83.2 to 89.8) is significantly lower (P < 0.01) compared with premenopausal women (success rate 95%, 95% CI 94% to 96%). CONCLUSION: Saline contrast hysterosonography is accurate in the evaluation of the uterine cavity in pre- and postmenopausal women suffering from abnormal uterine bleeding. The feasibility of saline contrast hysterosonography is high, although significantly better in premenopausal women compared with postmenopausal women. We conclude that saline contrast hysterosonography, in combination with an endometrium aspiration if necessary, can become the standard diagnostic procedure in women with abnormal uterine bleeding. PMID- 14550367 TI - Predicting residual disease after excision of cervical dysplasia. AB - Dysplastic epithelium at the resection margin after a cervical cone is known to predict persisting disease. We followed 702 women for 30 months after cervical excision to see which resection margin was predictive. The risk of persisting cytological abnormalities was doubled when CIN extended to the endocervical resection margin and was doubled when there was evidence of HPV. In contrast, disease at the ectocervical resection margin and the grade of CIN were not associated with a higher risk of residual disease. PMID- 14550368 TI - Massive obstetric haemorrhage resulting from a conservatively managed cervical pregnancy at delivery of its twin. PMID- 14550370 TI - Impact of rising caesarean rate on stillbirths in Merseyside. PMID- 14550371 TI - Rising caesarean section rates: a cause for concern? PMID- 14550373 TI - Outcomes of pregnancies in women with Type 1 diabetes in Scotland: a national population-based study. PMID- 14550376 TI - Development of innervation in primary incisors in the foetal period. AB - Sections from the frontal part of the mandible of 43 human foetuses from 9 to 39 weeks of prenatal age, which contained two, three and sometimes four lower incisors were immunohistochemically examined using protein gene product and neuron specific enolase (NSE) antibodies in order to establish the time of appearance of nerve fibres in the developing tooth germ and to define their topography. Nerve fibres were first detected in the dental follicle in the 11th week of intrauterine life. Their presence in the dental papilla was confirmed in the 18th week when the first layers of dentine and enamel were deposited. In the 24th week of intrauterine life, the nerve fibres first reached the subodontoblastic region. In the subsequent weeks, an increase in the number of nerve fibres accompanying blood vessels in the central portion of the dental papilla resulted in the formation of neuro-vascular bundles. Moreover, the progressive deposition of enamel and dentine was accompanied by branching of papillary nerves, which thereby formed a fan-pattern. In the foetal period, no evidence was found for the formation of a subodontoblastic plexus. However, we did observe single nerve fibres in close proximity to the odontoblast layer at the end of intrauterine life. Nerve fibres were not detected in either predentine or dentine throughout foetal life. PMID- 14550375 TI - Modelling oral malodour in a longitudinal study. AB - The aim of this study was to develop an in vitro model of the tongue microflora in order to assess anti-malodour compounds before clinical evaluation. Biofilms, derived from the tongue microflora, were grown in a constant depth film fermentor (CDFF) with nutrients supplied in the form of mucin- and serum-containing artificial saliva. Differential agars and a halimeter were used to determine the bacterial microflora and production of volatile sulphur compounds (VSCs), respectively. The resulting biofilms had a bacterial population which contained, on average, 29% streptococci, 48% Gram-negative anaerobes and 2.5% with an H2S producing phenotype. When the biofilms were pulsed with either chlorhexidine or zinc acetate there was a reduction in the number of H2S-producing bacteria, however these counts subsequently recovered as pulsing continued. The generation of VSCs was correlated to the viable counts of the H2S-producing bacteria. By pulsing with anti-malodour compounds over time we observed a reduction in the quantity of VSCs produced and a change in the composition of the plaque to one which contained fewer H2S-producing bacteria. PMID- 14550377 TI - Effect of mineral supplements to citric acid on enamel erosion. AB - The aim of this study was to evaluate the effect of mineral supplements to citric acid (1%; pH 2.21) on enamel erosion under controlled conditions in an artificial mouth. From each of 156 bovine incisors one polished enamel sample was prepared. The samples were divided among 13 experimental groups (n=12). In group 1 citric acid only was used (control). In groups 2-10 either calcium, phosphate or fluoride in various low concentrations was admixed to the citric acid. In groups 11-13 the citric acid was supplemented with a mixture of calcium, phosphate and fluoride. For demineralisation the specimens were rinsed with the respective solution for 1 min, immediately followed by a remineralisation period with artificial saliva (1 min). The specimens were cycled through this alternating procedure five times followed by rinsing for 8 h with artificial saliva. The de- and remineralisation cycle was repeated three times for each specimen interrupted by the 8 h-remineralisation periods. Before and after the experiments, the specimens were examined using microhardness testing (Knoop hardness) and laser profilometry. Hardness loss and enamel dissolution was significantly higher for the controls as compared to the remaining groups. Significantly lowest hardness loss for all groups was recorded for group 12 with admixture of calcium, phosphate and fluoride to citric acid. The significantly highest enamel loss was recorded for the controls compared to all other samples. Groups 3 and 4 revealed significantly lower and higher tissue loss compared to the remaining groups (2 13), respectively. The other groups did not differ significantly from each other. Modification of citric acid with calcium, phosphate and fluoride exerts a significant protective potential with respect to dental erosion. However, with the low concentrations applied enamel dissolution could not be completely prevented. PMID- 14550378 TI - A simple, yet accurate method for detecting and quantifying secretions from human minor salivary glands using the iodine-starch reaction. AB - The lack of published information about the minor salivary glands is due in part to the difficulties experienced in collecting and quantifying their secretions. In fact, no method exists for measuring their secretions that is both simple and accurate. This investigation examined the accuracy of our newly developed method (which simply employs the iodine-starch reaction) in 10 healthy non-medicated adults. A strip painted with a solution of iodine in absolute alcohol then with a fine starch powder mixed with castor oil was placed at a designated location on the lower-lip mucosa for 2 min to collect saliva. Black-stained spots of various sizes corresponding to the individual glands could be accurately visualized. After removal of the strip, the total stained area (mm2) was calculated by digitizing the spot areas using a computer system. The correlation coefficient (r) between known volumes of saliva and stain size was 0.995, indicating a close correlation. The correlation coefficient (r) between area values obtained in the first trial in each subject (Y) and the second (X; 10 min later) was 0.963, and the simple regression equation was close to Y=X, indicating good reproducibility. The mean flow rate microl/cm2 per min) obtained by converting mean total area to volume and thence to flow rate was 0.49+/-0.26, in good agreement with published values obtained by others. These results suggest that our newly developed method allows both the distribution and secretion rate of the minor salivary glands to be observed, and that it should be of practical value due to its simplicity, accuracy, and reproducibility. PMID- 14550379 TI - Neuromuscular objectives of the human masticatory apparatus during static biting. AB - OBJECTIVE: The central nervous system controls the muscles of mastication and may dictate muscle outputs according to a biologically important objective. This study tested the hypotheses that (a) the effective sagittal TMJ eminence morphology, and (b) the outputs of the masticatory muscles during static biting, are consistent with minimisation of joint loads or minimisation of muscle effort. DESIGN: Numerical modelling predicted effective eminence morphology (from sagittal plane directions of TMJ force for centred loading over a range from molar to incisor biting) and TMJ and muscle forces during static unilateral biting in seven subjects. In vivo effective eminence morphology was measured from jaw tracking recorded from each subject. Muscle activities during biting tasks on first molar and incisor teeth were measured by electromyography using surface or indwelling electrodes. RESULTS: Subject-specific predicted effective eminence morphology correlated with in vivo data (0.85< or =R2< or =0.99). Mixed and random coefficient analysis of covariance indicated good agreement between predicted and measured muscle outputs for all muscles of mastication investigated. Individual linear regression analysis showed that modelled muscle outputs accurately predicted EMG data, with average errors of 8% for molar and 15% for incisor biting. CONCLUSIONS: Effective sagittal eminence morphology was consistent with minimisation of joint loads for all subjects. Masticatory muscle outputs during unilateral biting were consistent with minimisation of joint loads or minimisation of muscle effort, or both, depending on the subject. These results are believed to be the first to test model predictions of muscle output during biting for all muscles of mastication. PMID- 14550380 TI - Functional significance of MMP-2 and MMP-9 expression by human malignant oral keratinocyte cell lines. AB - This study examined the expression of MMP-2 and MMP-9 in normal and human malignant oral keratinocytes. The expression of pro-MMP-2 and pro-MMP-9 was heterogeneous in the malignant cell lines. Normal oral keratinocytes expressed less pro-MMP-2 and more pro-MMP-9 than their malignant counterparts. Cells that expressed high levels of both MMP-2 and MMP-9 showed the greatest degree of invasion through Matrigel in vitro compared to cells with either low or variable levels of these enzymes; normal keratinocytes were non-invasive in these conditions. The degree to which the cells invaded through Matrigel was similar to their motility in the absence of Matrigel and was not influenced by the activation of the pro-enzymes or the inhibition of enzyme activity using a chemical inhibitor of gelatinases. Cells were transplanted orthotopically to athymic mice and demonstrated a variable capacity not only to form tumours at the site of inoculation but, also, to metastasise; normal oral keratinocytes were non tumorigenic. There was no correlation between the expression of either MMP-2 or MMP-9 and the tumorigenic/metastatic phenotype. The results emphasise the limitations of correlating in vitro and in vivo assays of tumour cell behaviour and suggest that invasion/motility in vitro may be a distinct phenotype from tumorigenicity/metastasis in vivo. PMID- 14550381 TI - The anatomy of buccinator--insights from functional casts of the oral vestibule. AB - OBJECTIVE: When a dental elastomer is placed in the oral vestibule and the facial muscles contracted while it sets, the functional cast produced frequently shows a deep anteroposterior groove, and marked posterosuperior and smaller anteroinferior bulges. This study investigates whether these features have a structural or a physiological basis. DESIGN: Casts of the right side of the oral vestibule of dissecting room cadavers and living volunteers were made using a polyvinylsiloxane dental elastomer. The volunteers each produced two functional casts in each of the following situations: while the teeth were clenched but the facial muscles inactive, while grimacing, and while swallowing. RESULTS: Grooves and bulges were largely absent in casts from the dissecting room cadavers and of living volunteers with the teeth clenched but the facial muscles inactive. They were present when the elastomer set while the subjects grimaced but most marked when they had been swallowing their saliva. The depth of the grooves varied between individuals and generally was greatest alongside the second molar teeth. We found that the posterosuperior bulge did not coincide with the position of the parotid duct. CONCLUSIONS: The anteroposterior groove represents a well developed horizontal part of buccinator. The regions of the casts bulging superior and inferior to the groove probably represent weaker regions of buccinator towards its maxillary and mandibular attachments. The bulge superior to the groove was not explained by a weakness in buccinator where it is pierced by the parotid duct. PMID- 14550382 TI - What is: disordered eating, amenorrhea, and osteoporosis? PMID- 14550383 TI - Exercise training, menstrual irregularities and bone development in children and adolescents. AB - Weight bearing physical activity plays an important role in bone development. This is particularly important in children and adolescents since bone mineral density reaches about 90% of its peak by the end of the second decade, and because about one quarter of adult bone is accumulated during the two years surrounding the peak bone growth velocity. Recent studies suggested that the exercise-induced increase in bone mineralization is maturity dependent, and that there is a "window of opportunity" and a critical period for bone response to weight bearing exercise during early puberty and premenarchal years. This supports the idea that increase in physical activity during childhood and adolescence can prevent bone disorders (like osteoporosis) later in life. In contrast, strenuous physical activity may affect the female reproductive system and lead to "athletic amenorrhea". The prevalence of "athletic amenorrhea" is 4 20 times higher than the general population. As a consequence, bone demineralization may develop with increased risk of skeletal fragility, fractures, vertebral instability, and curvature. Menstrual abnormalities in the female athlete result from hypothalamic suppression of the spontaneous pulsatile secretion of gonadotropin releasing hormone. Recent studies suggested that reduced energy availability (increased energy expenditure with inadequate caloric intake) is the main cause of the central suppression of the hypothalamic pituitary-gonadal axis. Therefore, effort should be made to optimize the nutritional state of female athletes, and if not successful, to reduce the training load in order to prevent menstrual abnormalities, and deleterious bone effects in particular during the critical period of rapid bone growth. PMID- 14550384 TI - A profile of adolescent females with a history of sexual assault in texas: familial environment, risk behaviors, and health status. AB - STUDY OBJECTIVE: To describe the prevalence and correlates of sexual assault among adolescent females. DESIGN: A cross-sectional study. SETTING: A university family planning clinic in south Texas. PARTICIPANTS: Female adolescents <18 years who initiated care at a university family planning clinic in south Texas between June 28, 1992, and April 28, 1994. The adolescents' lifetime sexual assault experience. MAIN OUTCOME MEASURES: Of the 791 adolescents interviewed, 167 (21%) reported a history of sexual assault. Sexual assault was found to be highly associated with multiple risky behaviors and depressive symptoms. Among assaulted adolescents, nonsexual risk behaviors (e.g., substance use) were more common among those who also experienced physical assault than among those who did not. Sexual risk behaviors (e.g., earlier age at sexual debut) and related gynecological infections (e.g., STDs) were more prevalent among women reporting forced sexual intercourse than in those who reported molestation only. Furthermore, adolescents assaulted by a stranger had participated in the most risky behaviors and reported the worst health status, while those assaulted by a date/acquaintance reported higher rates of inconsistent condom use and gynecological infections but lower rates of substance use and depressive symptoms than those assaulted by a family member. CONCLUSION: A deeper understanding of the links between the characteristics of assault, risk behaviors, and health conditions may provide opportunities to design more individualized interventions. PMID- 14550385 TI - WNT7A mutations in patients with Mullerian duct abnormalities. AB - STUDY OBJECTIVE: WNT7A gene mutations were evaluated as a potential cause for Mullerian duct derivative abnormalities in human females. The WNT gene family encodes glycoproteins that serve as signaling molecules during early development. The WNT7A gene has been previously identified as necessary for normal murine Mullerian duct development. WNT7A mutant mice display several Mullerian duct derivative abnormalities. DESIGN: Molecular genetic analysis of female patients with Mullerian duct derivative abnormalities. SETTING: Medical center-based academic research institution. PARTICIPANTS: 40 women with developmental abnormalities of the uterus and vagina and 12 normal controls. INTERVENTIONS: Polymerase chain reaction DNA amplification from human genomic DNA and denaturing gradient gel electrophoresis analysis of amplified DNA fragments. MAIN OUTCOME MEASURES: Presence or absence of WNT7A gene mutations in analyzed DNA fragments. RESULTS: No mutations were found in the WNT7A gene in any patient or control tested. CONCLUSIONS: WNT7A mutations are an unlikely cause of Mullerian duct derivative abnormalities in humans. PMID- 14550386 TI - Management of menstrual problems and contraception in adolescents with mental retardation: a medical, legal, and ethical review with new suggested guidelines. AB - The controversial history of the reproductive rights of the mentally retarded has led to the formulation of laws in the past century designed to protect women from forced sterilization. Significantly, however, in their official ethical guidelines, The American College of Obstetricians and Gynecologists states that "sterilization should not be denied to individuals simply because they also may be vulnerable to coercion" (Int J Gynaecol Obstet 1999; 65:317). Recent advances in medical and surgical methods of contraception and control of menstrual abnormalities have led to a re-evaluation of the management of adolescents with special needs. Physicians, the courts, parents, and caretakers need to be aware of the latest medical and surgical options available, the current applicable laws in each state if such exist, and the ethical guidelines to determine what treatment option is in the best interests of the patient. This review examines the history of the sterilization of the mentally retarded, the latest surgical and pharmacologic treatments available, and the current legal environment and proposes an algorithm to facilitate the management of menstrual hygiene and contraception. PMID- 14550387 TI - Acute abdominal pain with a calcified pelvic mass. AB - A case of a 15-year-old with a calcified pelvic mass is presented with a review of the differential diagnosis. PMID- 14550388 TI - Medical and other noncontraceptive uses of combined oral contraceptives. PMID- 14550389 TI - Management Quandary. Acute pelvic pain in adolescents. PMID- 14550390 TI - Examining pediatric and adolescent gynecology patients. PMID- 14550391 TI - Polycystic ovary syndrome. PMID- 14550394 TI - Differences between retinal and choroidal microvascular endothelial cells (MVECs) under normal and hypoxic conditions. AB - The morphological and functional differences between the retinal and choroidal vascular bed raise the question of whether the smallest functional unit, the microvascular endothelial cell (MVEC), also differs in its basal characteristics. Here, we examined bovine retinal and choroidal MVECs (rMVECs, cMVECs) for the presence and regulation of angiogenic mediators and their receptors, and cytokines at the mRNA level using quantitative RT-PCR and differential display. Vascular endothelial growth factor (VEGF) mRNA was expressed in both rMVECs and cMVECs. The basal and hypoxia-increased VEGF mRNA levels were significantly higher in cMVECs, which may indicate a higher capacity for autocrine stimulation in these cells. The mRNA for two VEGF receptors, Flt-1 and Flk-1, was present in rMVECs and cMVECs. Interestingly, rMVECs expressed higher Flt-1 but lower Flk-1 mRNA levels compared to cMVECs. Examining the angiopoietin (Ang)/Tie-2 system, we only detected Ang-1 mRNA at very low levels. While Ang-2 mRNA levels were high in both rMVECs and cMVECs, rMVECs expressed 2-3 times the basal and hypoxia upregulated Ang-2 mRNA levels than did cMVECs. No difference was found in basal Tie-2 mRNA levels. rMVECs are the more potent producers of macrophage colony stimulating factor (M-CSF) and granulocyte-macrophage CSF (GM-CSF), whereas cMVECs expressed higher RANTES mRNA levels. In our second approach - screening rMVECs and cMVECs for differentially expressed genes - we found liprin-beta1, calnexin, and sushi-repeat-containing protein, x chromosome (SRPX) mRNA in both MVEC types at varying levels. In summary, MVECs from the retinal and choroidal vascular beds showed quantitative differences in angiogenic regulator expression and in their capability to produce cytokines. PMID- 14550395 TI - Insulin-like growth factor-I (IGF-I) and transforming growth factor-beta (TGF beta) modulate tenascin-C and fibrillin-1 in bullous keratopathy stromal cells in vitro. AB - PURPOSE: Pseudophakic bullous keratopathy (PBK) is a major indication for corneal transplantation. Previous studies showed that PBK corneas had increased levels of insulin-like growth factor-I (IGF-I), bone morphogenetic protein-4 (BMP-4), transforming growth factor-beta (TGF-beta), interleukin-1alpha (IL-1alpha) and IL 8. The PBK corneas also had accumulations of tenascin-C (TN-C), fibrillin-1 (Fib 1), matrix metalloproteinase-2 (MMP-2), inflammatory cells but not myofibroblasts. Our goal is to determine if the growth factors/cytokines that are elevated in PBK corneas alter the expression of extracellular matrix (ECM) and/or degradative enzymes in vitro. METHODS: Stromal cell cultures from normal and PBK human corneas were established and treated for 6 days with IGF-I, BMP-4, IL 1alpha, IL-8 or TGF-beta1/beta2. Immunostaining, Western blot and dot blot analyses for TN-C, Fib-1, alpha-smooth muscle actin (alpha-SMA, a marker for myofibroblasts) or tissue inhibitor of metalloproteinase-1 (TIMP-1) were performed. RNAs were collected and analyzed with Northern blots for TN-C, Fib-1 and beta(2)-microglobulin. Culture media were analyzed using gelatin zymography with or without ethylenediaminetetraacetic acid (EDTA). Some samples were activated with p-aminophenylmercuric acetate (APMA) and reduction/alkylation, and the degradative activities were measured by the MMP-gelatinase activity assay kit. RESULTS: The IGF-I and TGF-beta1/TGF-beta2 increased (a) TN-C protein deposition, and (b) Fib-1 protein and RNA levels, but (c) had no significant affect on TIMP-1, matrix metalloproteinase-2 (MMP-2) or gelatinase activities. TGF-beta1/TGF-beta2 induced alpha-SMA protein (myofibroblasts) while IGF-I did not. BMP-4, IL-1alpha and IL-8 had little affect on the cells. CONCLUSIONS: Based upon our data, the fibrotic markers, TN-C and Fib-1, found in PBK corneas may be accounted for by IGF-I and TGF-beta. These growth factors promote fibrosis and ECM deposition without promoting proteolysis. While the other growth factors/cytokines are elevated in PBK corneas, their role(s) in PBK pathogenesis are not clear. In addition, exogenous IGF-I most closely elicited a response that was most similar to the characteristics of the PBK/ABK corneas, i.e. accumulation of TN-C and Fib-1 proteins in the absence of myofibroblasts. PMID- 14550396 TI - The photosensitiser xanthurenic acid is not present in normal human lenses. AB - UV light has often been investigated as a risk factor for the most common cause of blindness, human age-related cataract. One mechanism whereby UV light could induce cataract is via the action of photosensitisers. In this regard, xanthurenic acid has recently been highlighted since it has been reported to be present in the human lens and, in model studies, it markedly enhances the photo oxidation of proteins by wavelengths of light that penetrate the cornea. In this study we used HPLC and mass spectrometry to examine whether xanthurenic acid is indeed present in human lenses and, if so, the effect of age on its lenticular concentration. Xanthurenic acid could be formed artefactually by incubation of 3 hydroxykynurenine (3OHKyn) yellow, a known autoxidation product of the lenticular UV filter, 3OHKyn, in the presence of air and light, however, it could not be detected in any human lenses studied. Therefore, it appears unlikely that xanthurenic acid plays a role in lens aging or human cataract. PMID- 14550397 TI - Baseline characteristics of the transient pattern electroretinogram in non-human primates: inter-ocular and inter-session variability. AB - This study assessed the inter-ocular and inter-session variability of the transient pattern electroretinogram (PERG) in a group of non-human primates. The transient PERG was measured both eyes of 29 non-human primates, and again after three months in 23 eyes of 23 of these animals. Signals were elicited using a contrast (90%, 75 cdm(-2)) reversing (5 reversals sec(-1)) checkerboard pattern (0.56 cpd). PERGs were also measured for stimuli of varied spatial frequency (n=8, 0.07-2.22 cpd), contrast (n=4, 20-100%), mean luminance (n=4, 4.7-75 cdm( 2)) and defocus (n=5, +1, +2, +3 diopters). The inter-eye and inter-session limits-of-agreement (LOA; 95%) were determined for each PERG parameter. Variability was also compared with previous studies using the coefficient-of variability (COV). Pharmacological blockade of the inner retinal contributions to the PERG measured under these conditions was conducted in one animal using intravitreal injection of tetrodotoxin (approximately 6 microM) and N-methyl-D aspartic acid (approximately 6 microM). The N95 component of the primate transient PERG showed spatial tuning, with a peak between 0.14 and 0.28cpd. This spatial tuning was not as apparent for the P50 component. A linear relationship between P50 and N95 amplitude was found with contrast and mean luminance. Both components were attenuated with the introduction of +2 diopters or more of defocus. The inter-session COV for the P50 and N95 components were 23.8 and 19.2%, respectively, while the LOA were 58 and 46%, respectively. The N95:P50 ratio had smaller inter-session variability, was robust to changes in contrast, mean luminance and defocus, and was effective for characterization of inner retinal dysfunction after pharmacologic block. PMID- 14550398 TI - Insertion of MP20 into lens fibre cell plasma membranes correlates with the formation of an extracellular diffusion barrier. AB - It is known that during lens differentiation a number of fibre cell specific membrane proteins change their expression profiles. In this study we have investigated how the profiles of the two most abundant fibre cell membrane proteins AQP0 (formerly known as Major Intrinsic Protein, MIP) and MP20 change as a function of fibre cell differentiation. While AQP0 was always found associated with fibre cell membranes, MP20 was initially found in the cytoplasm of peripheral fibre cells before becoming inserted into the membranes of deeper fibre cells. To determine at what stage in fibre cell differentiation MP20 becomes inserted into the membrane, sections were double-labelled with an antibody against MP20, and propidium iodide, a marker of cell nuclei. This showed that membrane insertion of MP20 occurs in a discrete transition zone that coincided with the degradation of cell nuclei. To test the significance of the membrane insertion of MP20 to overall lens function, whole lenses were incubated for varying times in a solution containing either Texas Red-dextran or Lucifer yellow as markers of extracellular space. Lenses were fixed and then processed for immunocytochemistry. Analysis of these sections showed that both tracer dyes were excluded from the extracellular space in an area that coincided with insertion of MP20 into the plasma membrane. Our results suggest that the insertion of MP20 into fibre cell membranes coincides with the creation of a barrier that restricts the diffusion of molecules into the lens core via the extracellular space. PMID- 14550399 TI - Involvement of hepatic glucocorticoid receptor-mediated functions in steroid induced cataract formation. AB - Determination of whether the steroid-induced cataract formation is caused through glucocorticoid (GC) receptor-mediated process was conducted by using GC antagonist (RU486) and anti-GC receptor antibody, and by sucrose density gradient ultracentrifugation analysis. (1) When 15 day-old chick embryos were treated with dexamethasone (DEX, 0.025 micromol per egg), their lenses started to form an opaque ring around the peri-nuclear region (stage II-III) after 12 hr and developed into nuclear-like cataract (stage IV-V) after 44 hr. The cataract formation examined at the 44 hr could be effectively prevented by administration of RU486 (0.2 micromol per egg) ranging from 2 hr before to 12 hr after the DEX administration. (2) GC receptor was present in liver, but could not be determined in lens by western blot analysis using monoclonal anti-GC receptor antibody. (3) Sucrose gradient ultracentrifugation analysis indicated that the receptor (9S) in the liver could be transformed to the 4S form after 0.4M NaCl treatment. Combined with our previous data, this suggests that changes in hepatic functions mediated by the GC receptor after the GC administration may be involved in the process of the cataract formation. PMID- 14550400 TI - Modulation of cultured corneal keratocyte phenotype by growth factors/cytokines control in vitro contractility and extracellular matrix contraction. AB - The purpose of this study was to evaluate specific keratocyte phenotypes (keratocyte, fibroblast, myofibroblast) for cell contractility and ability to contract extracellular matrix. Rabbit keratocyte phenotype was modulated by exposure to optimal proliferative doses of IGF-I, IL-1alpha, FGF2, PDGF-AB, and TGFbeta(1). Cells were then evaluated by immunocytochemistry, western blot, collagen gel contraction and LPA stimulation to measure: (1) focal adhesion (FA), fibronectin (FN) and f-actin assembly; (2) expression of alpha-smooth muscle actin (alpha-SMA); (3) ability to contract extracellular matrix and (4) determine contractile ability, respectively. Untreated keratocytes showed no ability to contract collagen matrix. IGF-I and IL-1alpha increased cell proliferation (70.2 and 74.3%, respectively) but did not alter keratocyte phenotype or ability to contract matrix. FGF2 and PDGF induced fibroblast differentiation with FA and FN assembly and significant (p<0.05) extracellular matrix contraction. TGFbeta(1) induced myofibroblast differentiation with prominent FA and FN assembly, expression of alpha-SMA and significantly greater (p<0.05) matrix contraction. Addition of LPA induced actin filament assembly in growth factor starved fibroblasts and myofibroblasts but had no effect on the cultured keratocyte phenotype. We report for the first time that the keratocyte phenotype is non contractile and that cell quiescence is not a defining characteristic. We further establish that changes in environmental conditions modulate the keratocyte phenotype resulting in physiologically functional differences regarding cell contractility and capacity to contract extracellular matrix. PMID- 14550401 TI - Retinal pigment epithelium and endothelial cell interaction causes retinal pigment epithelial barrier dysfunction via a soluble VEGF-dependent mechanism. AB - PURPOSE: To investigate the effect of endothelial cells (EC) on the barrier function of the retinal pigment epithelium (RPE). METHODS: Primary bovine RPE were grown in solo culture or in coculture with bovine EC. Culture media of RPE were varied to develop a monolayer with stable barrier properties determined by transepithelial electrical resistance (TER) and permeability to sodium fluorescein. The effect of EC on the barrier properties of RPE was tested in contacting and non-contacting cocultures of RPE and EC. The conditioned media of cocultures were analysed for soluble vascular endothelial growth factor (VEGF) by ELISA. A neutralizing antibody to VEGF(165) was added to cocultures of RPE and EC and the TER was measured. RESULTS: RPE had maximal barrier properties (high TER, low permeability, positive staining for barrier proteins) at day 10 that persisted until day 20. Compared to solo RPE culture, cocultivation of RPE with EC reduced RPE barrier function significantly and led to a greater release of soluble VEGF into the conditioned media (p<0.05). Neutralizing VEGF with antibody led to partial recovery of barrier properties in the coculture conditions (p<0.03). CONCLUSIONS: Coculture of RPE with EC reduces RPE barrier properties and the reduction is, in part, mediated by soluble VEGF. EC-RPE contact-induced disruption of barrier properties occurs in ocular pathologies such as choroidal neovascularization, where EC move through Bruch's membrane and contact the RPE, leading to further exacerbation of the already compromised blood-retinal barrier. PMID- 14550403 TI - Effect of somatostatin and galanin on isolated rabbit iris sphincter and dilator muscles. AB - The neuropeptides somatostatin and galanin are present in the iris and may modulate pupil diameter. We examined the effects of somatostatin and galanin on isolated rabbit iris dilator and sphincter smooth muscles that were mounted in an organ bath. An isometric transducer recorded changes in tension in response to electric field stimulation (100 Hz, 0.3 msec in duration, 10 V in strength) delivered by a pair of platinum plate electrodes. The dilator muscle response to field stimulation was not changed by either peptide, even at the highest concentrations examined. The sphincter muscle response consisted of two components: a fast component mediated by acetylcholine and slow component mediated by substance P. Both somatostatin and galanin attenuated the cholinergic component in a dose-dependent manner (from 0.3 nM to 0.1 microM) but had no effect on responses mediated by substance P. Galanin was more effective (attenuation of 43% at 0.1 microM) compared with somatostatin (attenuation of 16% at 0.1 microM) in reducing the cholinergic response. Neither peptide affected the contraction induced by acetylcholine (1mM). Therefore both peptides inhibited cholinergic transmission in the sphincter muscle, although the degree of inhibition by each was different. We conclude that somatostatin and/or galanin may induce mydriasis by attenuating cholinergic neurotransmitter release. PMID- 14550402 TI - The matricellular protein SPARC is expressed in human trabecular meshwork. AB - PURPOSE: This investigation was undertaken to determine whether the matricellular protein SPARC is expressed in the human trabecular meshwork (TM) and cultured human trabecular meshwork cells. METHODS: Human donor trabecular meshwork and cultured cells obtained from trabecular meshwork were used in this study. Total RNA was obtained from TM and cultured TM endothelial cells, and RT-PCR was done with primers specific for SPARC. Western blotting was performed on donor TMs using an anti-SPARC monoclonal antibody prepared against rHuSPARC. Confocal microscopy was used to determine the distribution of SPARC in human anterior segments, and immunofluorescence on cultured TM cells was performed with the anti SPARC antibody. RESULTS: SPARC mRNA was expressed both in TM and in cultured TM cells. Immunoblotting for SPARC showed a doublet with a molecular mass approximately 43 kDa. The ratio of the doublet bands varied with each of the samples; some of the cultured cells and the tissue samples exhibited more of the upper band, and other cultured cells contained almost equal amounts of the two bands. The upper band was shown to be a glycosylated form of SPARC. Immunofluorescence showed that SPARC was expressed in the cultured TM, and confocal microscopy with the anti-SPARC antibody demonstrated the presence of this protein in the TM and in other tissues in the anterior segment. CONCLUSIONS: Our data conclusively show that SPARC mRNA and protein are present in non glaucomatous TM tissue and in cultured TM cells. Because of its effect on matrix metalloproteinases, SPARC may play a role in the regulation of intraocular pressure. PMID- 14550404 TI - Microtubule configuration and membranous vesicle transport in elongating fiber cells of the rat lens. AB - This study examines the microtubule configuration and its close association with the Golgi complex and Golgi-derived membranous vesicles in elongating fiber cells of the rat lens. Since fiber cells elongate tremendously during lens differentiation, we hypothesize that a microtubule-based motor system exists in the elongating fiber cells for transporting important membrane proteins and organelles to the target regions for cell growth. The newly synthesized membrane proteins are known to be transported from the trans-Golgi network in the form of vesicles to the target plasma membrane. By thin-section TEM, we observed a large number of vesicles of various sizes and shapes randomly distributed throughout the cytoplasm of elongating fiber cells. Both Golgi complex and vesicles exhibited characteristic normal structural features seen in other cell types and thus represented real vesicular organelles in the fiber cells. A large number of microtubules were regularly arranged into bundles parallel to the long axis of fiber cells as examined in both longitudinal and cross-section views. Many of these microtubules were closely associated or in intimate contact with the Golgi complex and vesicles in elongating fiber cells. The microtubule polarity assay revealed that microtubules exhibited a unidirectional polarity for the entire length of fiber cells as examined in both anterior and posterior cortical fiber segments. Namely, the minus end of microtubules was towards the anterior lens pole while the plus end was headed towards the posterior pole. This suggests that multiple molecular motors such as kinesin and dynein are needed for carrying the vesicles to both lens poles, since conventional kinesin is known to transport vesicular organelles towards the plus end whereas cytoplasmic dynein carries them towards the minus end of microtubules. By immunoblot analysis, we indeed detected the presence of both kinesin (120 kD) and dynein (70 kD) in homogenate prepared from lens cortical fibers. Moreover, immunogold TEM demonstrated that the aquaporin 0 (formally MIP26) antibody was localized on the membranous vesicles as well as plasma membranes of the cortical fiber cells. This study suggests that a microtubule-based motor system exists in the lens and plays an important role in transporting membrane proteins such as aquaporin 0 in the vesicles during fiber cell differentiation and elongation. PMID- 14550405 TI - Electroretinographic evidence for altered phototransduction gain and slowed recovery from photobleaches in albino mice with a MET450 variant in RPE65. AB - Our purpose was to investigate the physiological phenotype of albino mice with a variation in the Rpe65 gene encoding either methionine or leucine at amino acid #450. Full-field electroretinograms (ERGs) were recorded from C57BL/6J-c(2J) albino mice with MET450 and BALB/cByJ albino mice with LEU450. Recordings from pigmented mice (C57BL/6J) served as controls. Rod ERG a-waves were fitted with a model to estimate parameters of activation. Recovery of function following a photobleach was studied by monitoring the return to pre-bleach a- or b-wave amplitudes of the dark-adapted electroretinogram. The parameter, S, derived from the fit of the rod model, was significantly higher for albino mice compared to pigmented controls. Between the albino mice, S was highest for BALB/cByJ compared to C57BL/6J-c(2J). The parameters t(d) and Rm(P3) were not different across the three strains. The difference in S between the BALB/cByJ and C57BL/6J-c(2J) albino strains is interpreted to reflect differences in intrinsic phototransduction gain. Recovery from a photobleach was also slower for the C57BL/6J-c(2J) albino mice compared with BALB/cByJ albino mice, consistent with prior studies showing slowed rhodopsin regeneration in mice with the RPE65 METH450 variant. ERG recordings show that C57BL/6J-c(2J) albino mice with the MET450 variant of the RPE65 protein have a lower gain of activation and slower recovery from photobleach than do the BALB/cByJ albino mice with LEU450. Both the slower recovery from photobleach and lower gain of activation characteristic of the C57BL/6J-c(2J) strain may contribute to the mechanism by which it is protected from light-induced photoreceptor death relative to BALB/c. PMID- 14550406 TI - Optical development in the foetal bovine lens. AB - The refractive index variations along the optic axes of nine foetal bovine lenses were measured using an optic fibre reflectometer. The device measures refractive index directly, using the principle of Fresnel reflectance. These results show that lenses with a wet weight >0.7 g (corresponding to 4.5 months gestational age), have a refractive index profile which approximates a parabolic shape, akin to that found in post-natal and adult bovine lenses. In lenses of wet weight below 0.7 g, the refractive index distributions show irregularities. These findings suggest that the refractive index profile in the bovine lens starts to form into a parabolic distribution, as seen in adult lenses, about half-way through gestation. The trigger for this formation is not known. PMID- 14550408 TI - Gene repression by Polycomb group protein complexes: a distinct complex for every occasion? AB - Polycomb group (PcG) proteins play important roles in maintaining the repressed transcriptional state of genes. PcG proteins operate as part of Polycomb repressive complexes (PRCs). 'Core' PRCs have been purified that contain only a limited number of PcG proteins. In addition, many gene regulatory proteins have been identified to interact with PcG proteins. However, it remains subject to discussion whether these interactions are transient or whether the regulatory proteins are real components of PRCs. It has also become clear that the compositions of 'core' PRCs differ amongst cell types and that extensive changes in compositions occur during the embryonic development of cells. Because of these dynamic changes, we argue that speaking of a definitive core PRC can be misleading. PMID- 14550409 TI - Chromatin regulation during C. elegans germline development. AB - Recent studies in Caenorhabditis elegans implicate PcG- and NuRD-like chromatin regulators in the establishment and maintenance of germline-soma distinctions. Somatic cells appear to utilize NuRD-related nucleosome-remodeling factors to overwrite germline-specific chromatin states that are specified through PcG-like activities. The germline, in turn, may rely on an asymmetrically inherited inhibitor to prevent chromatin reorganization that would otherwise erase pluripotency. PMID- 14550410 TI - Turning germ cells into stem cells. AB - Primordial germ cells (PGCs), the embryonic precursors of the gametes of the adult animal, can give rise to two types of pluripotent stem cells. In vivo, PGCs can give rise to embryonal carcinoma cells, the pluripotent stem cells of testicular tumors. Cultured PGCs exposed to a specific cocktail of growth factors give rise to embryonic germ cells, pluripotent stem cells that can contribute to all the lineages of chimeric embryos including the germline. The conversion of PGCs into pluripotent stem cells is a remarkably similar process to nuclear reprogramming in which a somatic nucleus is reprogrammed in the egg cytoplasm. Understanding the genetics of embryonal carcinoma cell formation and the growth factor signaling pathways controlling embryonic germ cell derivation could tell us much about the molecular controls on developmental potency in mammals. PMID- 14550411 TI - Transdetermination in Drosophila imaginal discs: a model for understanding pluripotency and selector gene maintenance. AB - Drosophila imaginal disc cells have the ability to undergo transdetermination, a process whereby determined disc cells change fate to that of another disc identity. For example, leg disc cells can transdetermine to develop as wing cells. Such events can occur after mechanical disc fragmentation and subsequent regeneration. A subset of transdetermination events can be induced in situ by misexpression of the signaling gene wingless. Both fragmentation and wingless induce transdetermination by altering the expression of selector genes, which drive disc-specific developmental programs. An important future goal is to address how signaling pathways interact with chromatin structure to regulate and maintain the proper expression of selector genes. PMID- 14550412 TI - Cell fusion: an alternative to stem cell plasticity and its therapeutic implications. AB - Cell fusion has long been known to produce viable cells and to have a major role in mammalian development and differentiation. As gene expression profiles can change after cell fusion, this event must be also considered as an alternative explanation for the many cases of 'stem cell plasticity' that have been recently described and are promoted as a promising therapeutic strategy. Cell fusion has been demonstrated to occur in some recent studies, and the available evidence is often not inconsistent with cell fusion in others. Cell fusion itself has therapeutic potential, but low rates of spontaneous fusion and safety concerns may ultimately limit its use. PMID- 14550413 TI - Reprogramming in nuclear transfer. AB - Reprogramming a nucleus by transferring it to oocyte cytoplasm triggers epigenetic changes that eventually lead to the restoration of a totipotent state and the birth of a viable animal. A simplified way of studying this complex process is to study cell hybrids. These studies suggest that the pluripotent character of one nucleus is dominant over that of the other nucleus. The development of nuclear transfer embryos shows that there are several restriction points and that the extra-embryonic lineages may be the primary source of death. Successful reprogramming depends on the balance between epigenetic modifications and the regulative properties of development. PMID- 14550414 TI - Regeneration and gene regulation in planarians. AB - Planarians can regenerate using a pluripotent stem cell system. This phenomenon provides a unique opportunity to understand gene regulation in the process of differentiation from pluripotent stem cells. Recent studies have made significant advances in our understanding of the pluripotent stem cell system in this model. In particular, a gene knockdown method by RNA interference enabled great progress in identifying genes involved in regeneration and stem cell regulation. PMID- 14550415 TI - Cell differentiation and cell fate during urodele tail and limb regeneration. AB - One of the most striking natural examples of adult tissue plasticity in vertebrates is limb and tail regeneration in urodele amphibians. In this setting, amputation triggers the destabilization of cell differentiation and the production of progenitor cells that extensively proliferate and pattern themselves to recreate a perfect replica of the missing part. A precise understanding of which cells dedifferentiate and how plastic they become has recently begun to emerge. Furthermore, information on which developmental gene programs are activated upon injury is becoming better understood. These studies indicate that, upon injury, an unusual cohort of genes are co-expressed. The future challenge will be to link the systems for studying dedifferentiation with activation of gene expression to understand on a molecular level how cells are 'pushed backward' to regenerate a complex structure such as a limb or tail. PMID- 14550416 TI - Tissue generation from amphibian animal caps. AB - Formation of three germ layers is the most important event in early vertebrate development. Animal cap assays can be used to reproduce the in vivo induction of amphibian tissues in order to investigate the differentiation processes that occur in normal embryonic development. Activin treatment strongly and dose dependently induces various types of mesodermal and endodermal tissue in cultured animal caps. Beating heart, pronephros, pancreas and cartilage can be induced by microsurgical manipulation and simultaneous treatment with activin and other factors. These in vitro induction systems will be helpful for elucidating the mechanisms of tissue induction and organ formation in vertebrate development. PMID- 14550417 TI - Self-renewal and lineage restriction of hematopoietic stem cells. AB - Over the past decade, the purification and characterization of hematopoietic stem cells have ascertained their presence at the clonal level although they had hitherto existed conceptually. Now we have begun to understand their functions in molecular terms. Several important works indicative of such a new era in stem cell biology have been published recently. In particular, Bmi1, which belongs to the Polycomb group of genes, has been implicated as one of the basic molecules to maintain the proliferation capacity in hematopoietic stem cells. We need to seek other similarly important molecules for their functions. Perhaps studying interactions among genes is one of the most exciting subjects in stem cell research. PMID- 14550418 TI - The emergent design of the neural tube: prepattern, SHH morphogen and GLI code. AB - The Sonic hedgehog (Shh) pathway plays an important role in the development of many tissues and organs. The secreted ligand Shh has been shown to act as a mitogen, morphogen and survival factor in different contexts whereas the three Gli transcription factors act as Shh mediators in a context-dependent combinatorial fashion. The common wisdom has been that Gli protein function is subject to Shh signaling. One can ask how Gli proteins act and what the nature of Shh signaling during CNS dorsal-ventral patterning is. Is it possible that Hedgehog signals are only one of several ways to regulate Gli activity? Moreover, in light of the partial rescue of the neural tube phenotype of Shh or Smoothened mutant embryos in Shh(-/-);Gli3(-/-), Smoothened(-/-);Gli3(-/-), and Shh(-/ );Rab23(-/-) double null embryos, one can consider the roles that the Shh-Gli pathway may have taken to orchestrate congruent prepattern and growth, and the importance of creating the correct number of precursors in patterning mechanisms. PMID- 14550419 TI - Transcription factors controlling the beginning and end of B-cell differentiation. AB - Powerful techniques--including gene targeting, genomic and proteomic analyses- are allowing a more complete understanding of the transcription factors that drive B-cell development. Regulatory cascades involving multiple steps have been identified at many decision points. In addition, feedback loops and transcriptional inhibition of alternative fates have been found to be important both for commitment to the B lymphocyte lineage and for terminal B-cell differentiation. PMID- 14550420 TI - Multipotentiality of the neural crest. AB - Multiple neural and non-neural cell types arise from the neural crest (NC) in vertebrate embryos. Recent work has provided evidence for multipotent stem cells and intermediate precursors in the early NC cell population as well as in various NC derivatives in embryos and even in adult. Advances have been made towards understanding how cytokines, regulatory genes and cell-cell interactions cooperate to control commitment and differentiation to pigment cells, glia and neurone subtypes. In addition, NC cell fates appeared to be unstable, as differentiated NC cells can reverse to multipotent precursors and transdifferentiate in vitro. PMID- 14550421 TI - Mesoangioblasts--vascular progenitors for extravascular mesodermal tissues. AB - Mesoangioblasts are multipotent progenitors of mesodermal tissues that express the key marker of angiopoietic progenitors, Flk1 (VEGF-receptor 2), and are physically associated with the embryonic dorsal aorta in avian and mammalian species. When transplanted in vivo, they give rise to multiple differentiated mesodermal phenotypes. Their ability to extensively self-renew in vitro, while retaining multipotency, qualifies mesoangioblasts as a novel class of stem cells. Mesoangioblasts disclose not only an unexpected source of progenitors for skeletal muscle and a variety of other mesoderm-derived tissues, but also establish a lineage kinship between progenitors of vascular and extravascular mesodermal tissues, with important basic and applicative implications. PMID- 14550422 TI - A niche for adult neural stem cells. AB - The adult mammalian brain harbors multipotent stem cells, which reside and participate in specialized niches that support self-renewal and differentiation. The first cellular and molecular elements of the stem cell niche in the adult brain have been identified and include cell-cell interactions and somatic cell signaling, the vasculature, the extracellular matrix and basal lamina. Furthermore, regulation at the epigenetic level via chromatin modification and remodeling is an integral aspect of stem cell biology. Understanding the in vivo stem cell niche will provide a framework for the elucidation of stem cell function in the adult brain. PMID- 14550423 TI - Plant stem cells: divergent pathways and common themes in shoots and roots. AB - Stem cells in plant shoot and root meristems are maintained throughout the life of the plant and produce somatic daughter cells that make up the body of the plant. Plant stem cells can also be derived from somatic cells in vivo and in vitro. Recent findings are refining our knowledge of signaling pathways that define stem cell fate and specify either shoot or root stem cell function. New evidence also highlights a role for epigenetic mechanisms in controlling stem cell fate. PMID- 14550424 TI - Indications for early cystectomy. PMID- 14550425 TI - Measuring self-reported sexual function in men with prostate cancer. PMID- 14550426 TI - Molecular genetic aspects of prostate transition zone lesions. PMID- 14550427 TI - Levofloxacin for chronic prostatitis/chronic pelvic pain syndrome in men: a randomized placebo-controlled multicenter trial. AB - OBJECTIVES: To perform a Canadian multicenter randomized placebo-controlled trial to evaluate the safety and efficacy of 6 weeks of levofloxacin therapy compared with placebo in chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). Uncontrolled studies have supported the use of antibiotics in CP/CPPS. METHODS: Men with a National Institutes of Health (NIH) diagnosis of CP/CPPS (specifically, no infection localized to the prostate) were randomized to levofloxacin (500 mg/day) or placebo for 6 weeks in 11 Canadian centers. Patients were assessed at baseline and at 3, 6, and 12 weeks with the NIH Chronic Prostatitis Symptom Index (NIH-CPSI) and global patient assessments (subjective global assessment and patient assessment questionnaire). RESULTS: Eighty men (average age 56.0 years, range 36 to 78; duration of symptoms 6.5 years, range 0.6 to 32) were randomized to receive levofloxacin (n = 45) or placebo (n = 35). All were evaluated in an intent-to-treat analysis. Both groups experienced progressive improvement in symptoms as measured by the NIH-CPSI. However, the difference in response was not statistically or clinically significant at end of treatment (6 weeks) or at the end of the follow-up visits (12 weeks). No patients withdrew because of adverse events. One patient withdrew before the 6-week assessment. Adverse events (all mild) were reported in 20% of the levofloxacin group and 17% of the placebo group. CONCLUSIONS: This pilot placebo-controlled study showed that 6 weeks of levofloxacin therapy in men diagnosed with CP/CPPS resulted in symptom improvement that was not significantly different from that with placebo at end of treatment or follow-up. The clinical ramifications of these findings need to be addressed. PMID- 14550428 TI - Identification of common themes from after-hour telephone calls made to urology residents. AB - OBJECTIVES: To characterize the content and sources of after-hour telephone calls from a general urology practice so that common themes can be identified and incorporated into the education of urology residents. METHODS: After-hour telephone calls were prospectively assessed for 6 months by a single senior urology resident. Calls occurring on weekends/holidays and between 5 PM and 8 AM on weekdays were directly referred to the on-call resident. The day of week, time, source, initiating event, and required action from each call was recorded. Exclusion criteria included calls regarding established hospital urology in patients and duplicate calls from individual patients calling more than once within 24 hours. RESULTS: Eighty-seven calls were collected. Seventy percent occurred on weekends, primarily between 8 AM and 5 PM. The most common source was from outpatients (56%) followed by emergency room physicians (30%). In general, 61% and 26% of all calls regarded new urologic symptoms and postoperative issues, respectively. Urinary calculi were the most common specific reason for a call, followed by lower urinary tract symptoms and hematuria. An acute urologic evaluation was required for 27% of all telephone calls. Nine patients (10%) required admission within 48 hours of calling. CONCLUSIONS: The education of urology residents should emphasize telephone evaluations of patients who present with urinary calculi and common postoperative issues. The number of after-hour calls may be able to be reduced if patients and nonurologist physicians are also educated in the management of nonacute urologic problems. PMID- 14550429 TI - Management of encrusted ureteral stents impacted in upper tract. AB - OBJECTIVES: To present our series of patients with ureteral stent encrustation and give indwelling times and management. Encrustation is one of the most serious complications of ureteral stents. METHODS: A retrospective review was undertaken of all encrusted stents during a 4-year period. The inclusion criterion was a stent that required some form of intervention above the ureteral orifice to remove it. Combinations of extracorporeal shock wave lithotripsy, ureteroscopy, percutaneous nephrolithotomy, and open surgery were used to achieve stent removal. RESULTS: Forty-nine impacted encrusted stents were treated in 41 patients. Of these, 75.5% had become encrusted within 6 months and 42.8% within 4 months. The mean indwelling time was 5.6 months. Forty-seven stents were removed by endourologic techniques, with 4 requiring extracorporeal shock wave lithotripsy alone, 28 ureteroscopy, and 10 a combination of both. Five patients underwent successful percutaneous nephrolithotomy. One patient underwent open surgery, and in one removal failed. The mean number of procedures per patient was 1.94. CONCLUSIONS: Stent encrustation can pose a serious challenge to the endourologist, and indwelling times should be minimized to avoid problems. Patients often require multiple treatments and a combination of extracorporeal shock wave lithotripsy and ureteroscopy offers highly successful outcomes and often avoids the need for more invasive techniques. PMID- 14550430 TI - Laparoscopy for the diagnosis and treatment of radiologically occult but symptomatic hypoplastic kidneys. AB - OBJECTIVES: To evaluate the efficacy of different imaging modalities for visualization of small poorly functioning hypoplastic and dysplastic kidneys and to assess the role of laparoscopy in localization and treatment. METHODS: Between 1998 and 2002, 10 female patients who presented with urinary incontinence, flank pain, or hypertension secondary to small, poorly functioning hypoplastic or dysplastic kidneys were treated at our institute. We reviewed the results of the imaging studies, cystoscopy, and evaluation under anesthesia for these patients. After the diagnosis, all patients were treated with laparoscopic retroperitoneal or transperitoneal nephroureterectomy or nephrectomy. RESULTS: Intravenous urography and ultrasonography failed to visualize the affected renal units in all 10 cases. Computed tomography could locate small dysplastic renal units in only 5 patients (50%). Dimercaptosuccinic acid renal scintigraphy was diagnostic in all 10 cases (100%). Magnetic resonance urography was done in 2 cases only, and the affected renal units were identified in both. Cystoscopy and vaginoscopy were inconclusive in all but 2 cases. Laparoscopy efficiently located the offending renal units in all 10 cases. In 9 cases, the affected renal units were located in the lumbar region, and in 1 case it was situated ectopically in the pelvis. All 10 patients underwent laparoscopic retroperitoneal or transperitoneal nephroureterectomy or nephrectomy and were asymptomatic after surgery. CONCLUSIONS: Dimercaptosuccinic acid scintigraphy is an accurate and specific imaging modality for visualization of small hypoplastic renal units that are not visualized by conventional radiologic imaging techniques. Laparoscopy can be recommended as the ideal method of management, because it provides a minimally invasive technique for detecting the kidney and simultaneously correcting the problem. PMID- 14550431 TI - Modified renal morcellation for renal cell carcinoma: laboratory experience and early clinical application. AB - OBJECTIVES: To present our modified technique to expedite the entrapment and morcellation process. Standard renal morcellation is typically performed using a 12-mm incision, with the specimen entrapped within a durable LapSac. Because the LapSac does not have a deployment mechanism, its application is technically demanding and time consuming. The recent elimination of the electrical morcellator has also made the morcellation process time consuming and tedious. METHODS: Using an in vitro porcine kidney morcellation model, we compared traditional morcellation (n = 10) with our modified morcellation technique (n = 10). The modified morcellation technique incorporated an EndoCatch entrapment sac and used standard manual morcellation through a 3-cm incision. The 3-cm incision allowed for extracorporeal morcellation, because the specimen could be seen through the abdominal wall defect. Subsequently, we performed 11 laparoscopic radical/total nephrectomies for renal masses using the modified morcellation technique through a 3-cm incision. RESULTS: In this in vitro model, renal specimen morcellation with the standard technique required a mean of 9.4 minutes (16.5 g/min) and modified morcellation required 3.24 minutes (51.24 g/min; P <0.0001). The mean fragment size with standard and modified morcellation was 1.47 and 4.46 g, respectively (P <0.0001). EndoCatch entrapment sack integrity was confirmed in all 10 cases (100%). Clinically, 12 patients underwent modified morcellation of radical nephrectomy specimens. All specimens were renal cell carcinoma, with a mean specimen weight of 724 g. The mean entrapment time was 2 minutes, 40 seconds, and the mean morcellation time was 11 minutes. EndoCatch entrapment sack integrity was confirmed in all cases. At a mean follow-up of 8 months, no trocar site or local recurrences have developed. CONCLUSIONS: Preliminary laboratory and clinical data have demonstrated that the modified morcellation technique is safe and expeditious. The larger fragments that result from the modified technique may facilitate tumor staging. The technique must be performed with great care to prevent sack perforation. Prospective randomized comparison of the modified and standard morcellation techniques and long-term follow-up are required before routine application of this technique. PMID- 14550433 TI - Nephrectomy for metastatic renal cell carcinoma: Indiana University experience. AB - OBJECTIVES: To report on the short-term morbidity of radical nephrectomy in 32 patients with poorer performance status and more advanced primary renal cell carcinoma (RCC). Nephrectomy followed by immunotherapy has been shown to improve survival in selected, good performance status patients with metastatic RCC. METHODS: We report on 32 patients who underwent radical nephrectomy (20 open procedures and 12 laparoscopic) in the setting of metastatic RCC at Indiana University between 1999 and 2002. The study group included patients with advanced primary tumors (inferior vena cava involvement, large size, and involvement of adjacent structures). The patients' performance status score ranged from 0 to 2. RESULTS: The average hospital stay was 5.1 days. No significant intraoperative complications were encountered, and postoperative complications occurred in 6 patients, including one perioperative death. At 4 weeks postoperatively, 21 (72.4%) of 29 assessable patients had a performance status equal to, or better than, their preoperative status, including 4 patients who converted from a preoperative performance status of 2 to 0 or 1 postoperatively. Eleven patients (34.4%) went on to receive postoperative immunotherapy. CONCLUSIONS: The results of our study demonstrated that radical nephrectomy in the setting of metastatic RCC has a low morbidity and acceptable recovery in these patients with advanced primary tumors and poorer performance status. PMID- 14550434 TI - Nephron-sparing surgery for renal cell carcinoma: clinicopathologic features predictive of patient outcome. AB - OBJECTIVES: To determine the clinicopathologic features associated with outcome in patients with sporadic renal cell carcinoma (RCC) treated with nephron-sparing surgery. METHODS: We studied 344 patients treated with nephron-sparing surgery between 1970 and 2000. The pathologic features of the tumors were reviewed by two urologic pathologists who recorded the histologic subtype, 2003 TNM stage, tumor size, and grade. Cancer-specific survival (CSS) was estimated using the Kaplan Meier method, and log-rank tests were used to compare the outcome by histologic subtype. Univariate Cox proportional hazards models were fit to assess the associations between the clinicopathologic features and death from RCC, distant metastases, and local recurrence. RESULTS: The CSS rate at 5 and 10 years for patients with clear cell RCC was 94.4% and 91.5%, respectively. In contrast, the CSS rate at 5 and 10 years for patients with papillary or chromophobe RCC was 99.0%, because only 1 patient died of papillary RCC and no patient died of chromophobe RCC (P = 0.029). Among the patients with localized clear cell RCC, tumor stage and grade were significantly associated with death from RCC and metastases. Grade was significantly associated with local recurrence for clear cell RCC, but not for papillary RCC. CONCLUSIONS: In our series of patients with RCC treated with nephron-sparing surgery, patients with clear cell RCC had a significantly worse CSS than did patients with papillary and chromophobe RCC. Tumor stage and grade were associated with outcome among patients with localized clear cell RCC. These findings are similar to the results for patients with localized clear cell RCC treated with radical nephrectomy. PMID- 14550435 TI - Detrusor resistive index evaluated by Doppler ultrasonography as a potential indicator of bladder outlet obstruction. AB - OBJECTIVES: To appraise detrusor blood flow by Doppler ultrasonography in men with suspected bladder outlet obstruction (BOO) to determine whether this imaging technique provides useful information for the assessment of BOO. Experimental studies have shown that BOO is associated with reduced blood flow to the detrusor. METHODS: Twenty-nine consecutive men with lower urinary tract symptoms were prospectively enrolled. A urodynamic pressure-flow study was performed by the urologist to determine BOO, and Doppler ultrasonography was subsequently performed by the radiologist. The physicians were unaware of the other's results. Scanning was performed on a filled and empty bladder. Arterial blood flow was measured at three distinct sites, the two lateral walls and the trigone, and the resistive index (RI) of each site was calculated (RI = (V(MAX) - V(MIN))/V(MAX)). For each patient, the arithmetic average of the three RIs was defined as the detrusor RI. The findings were compared between patients with and without evidence of BOO. A logistic regression model tested the predictive value of the RI. RESULTS: According to the pressure-flow study results, 22 (75%) and 7 (25%) of the 29 patients were diagnosed as having or not having BOO, respectively. A statistically significant difference was found between the detrusor RI in the obstructed versus nonobstructed patients in both full (P <0.001) and empty (P <0.03) bladder states (0.79 versus 0.68 and 0.74 versus 0.66, respectively). Our logistic regression model predicted BOO with an overall accuracy of 86%, positive predictive value of 95%, and negative predictive value of 57%. CONCLUSIONS: The RI of arterial blood flow in the detrusor measured by Doppler ultrasonography provides important predictive information for the presence of BOO. Additional studies are warranted to validate our results and explore the role of Doppler ultrasonography in the management paradigms of patients with suspected BOO. PMID- 14550436 TI - Distinguishing neurogenic from non-neurogenic detrusor overactivity: a urodynamic assessment of lower urinary tract symptoms in patients with and without Parkinson's disease. AB - OBJECTIVES: To examine the urodynamic (UDS) attributes of detrusor overactivity (DO) in patients with Parkinson's disease in comparison to DO in men without neurologic disease, in whom DO is presumably outlet obstruction induced. METHODS: The UDS database was reviewed for three groups of patients: group 1, men with lower urinary tract symptoms (LUTS) and no known neurologic condition with DO (n = 22); group 2, men with Parkinson's disease and LUTS (n = 39); and group 3, women with Parkinson's disease and LUTS (n = 18). Statistical analysis was used to compare the UDS parameters and diagnoses among the groups and to test for associations between Parkinson's disease duration, Hoehn and Yahr score, and UDS findings. RESULTS: Patients with Parkinson's disease had a significantly lower median volume at first detrusor contraction than those with non-neurogenic DO. The percentage of group 1 patients with urge incontinence was significantly lower than that found in the other two groups (9.1% versus 53.8% and 55.6%, P <0.001 and 0.002, respectively). No statistically significant correlation between the duration or severity of Parkinson's disease and UDS parameters was found. CONCLUSIONS: Men with non-neurogenic LUTS are less likely to have urge incontinence on UDS than either men or women with Parkinson's disease. DO owing to Parkinson's disease occurs earlier during filling compared with non-neurogenic DO, especially in women. The duration and severity of Parkinson's disease are not predictive of the nature or severity of UDS abnormalities. PMID- 14550437 TI - Accuracy of bladder volume determinations by ultrasonography: are they accurate over entire bladder volume range? AB - OBJECTIVES: To investigate the accuracy of a portable three-dimensional hand-held BladderScan (BS) and two-dimensional conventional ultrasonography (CUS) in the estimation of bladder volumes. METHODS: The bladder volume of 65 subjects was measured during filling cystometry using two ultrasound modalities. Serial measurements were performed when the infused volume reached 100, 200, 300, and 400 mL; the investigator measured the volumes three times each with BS and CUS. Each corresponding true volume was calculated by interpolation using zero volume and the final catheterized volume at the end of cystometry. The accuracy of the two ultrasound methods was compared using raw scaled values of the volumes and the percentage of differences of volume. RESULTS: Bladder volumes determined by CUS and BS were underestimated by 21.8% and 3.3% (mean values), respectively. The percentage of differences of volume of BS was not statistically significant (P >0.05), but that of CUS was statistically significant (P <0.00001). No significant percentage of differences of volume change according to the volume status was observed by either ultrasound measurement method (P >0.05). Linear regression analysis between the true volumes and the raw scaled volumes showed that the slope of the BS was closer to 1.0 than that of the CUS, and the difference was statistically significant (P <0.001). The effects of diagnosis, age, sex, and body mass index were not significant in determining the bladder volume by either method (P >0.05). CONCLUSIONS: Our results demonstrated that a three-dimensional hand-held scanner measures the bladder volume in a reproducible and accurate manner for a wide range of bladder volumes and is superior to two dimensional stationary ultrasonography. PMID- 14550438 TI - Reducing time to oral diet and hospital discharge in patients undergoing radical cystectomy using a perioperative care plan. AB - OBJECTIVES: To outline our current perioperative treatment of patients undergoing radical cystectomy and urinary diversion, which uses advancements in perioperative care to allow for early institution of an oral diet and early hospital discharge, and thereby overall improvement in patient recovery and outcome after this procedure. METHODS: Forty consecutive patients underwent radical cystectomy and urinary diversion with curative intent from 2001 to 2002. A care plan was followed for all patients and included improvements in preoperative, intraoperative, and postoperative care. The preoperative care included limited outpatient bowel preparation with sodium phosphate solution and patient education. Operative modifications included reduced incision length, initial preperitoneal dissection, and the use of internal surgical stapling devices. The postoperative care included the use of prokinetic agents, early nasogastric tube removal, the use of non-narcotic analgesics, and early institution of an oral diet. The outcomes with regard to time to institution of an oral diet, tolerance of a regular diet, and hospital discharge were assessed. RESULTS: The mean surgical time was 3.9 hours, and the mean estimated blood loss was 573 mL. The mean time to the institution of a clear liquid diet was 2.0 days and to a regular diet was 4.2 days. The mean time to hospital discharge was 5.1 days. No statistically significant differences were found in the time to resumption of a regular diet or to discharge between patients undergoing ileal conduits versus orthotopic ileal neobladders. Only 1 patient had any gastrointestinal dysfunction (ileus), and this patient was discharged on postoperative 7. No patient had any delayed complications involving problems with diet intolerance or other gastrointestinal dysfunction. The results of the current series were compared with those of historical controls. CONCLUSIONS: Advancements in preoperative, intraoperative, and postoperative management have together been successfully used in our patient population to reduce morbidity and improve recovery with regard to the early institution of an oral diet and early hospital discharge. PMID- 14550440 TI - Weak correlation between bladder outlet obstruction and probability to void to completion. AB - OBJECTIVES: To investigate the weak correlation between bladder outlet obstruction (BOO), as diagnosed using the provisional International Continence Society nomogram for the definition of BOO in men, and postvoid residual urine volume. METHODS: The relationship between voiding to completion and several indexes for bladder outlet resistance and bladder contractility was studied in 131 pressure flow studies in male patients using multivariate logistic regression analysis. RESULTS: The International Continence Society nomogram and the related BOO index weakly predict for postvoid residual urine volume (areas under the receiver operating characteristic curve 0.63 and 0.64, respectively). The BOO index primarily measures bladder outlet resistance. If the nomogram or BOO index is augmented with bladder contractility information, the postvoid residual urine volume can be predicted significantly better (eg, area under the receiver operating characteristic curve [0.89] for the combination of the BOO index and bladder contractility information). CONCLUSIONS: The weak correlation between BOO and postvoid residual urine volume is related to the fact that emptying the bladder to completion depends on bladder contractility, as well as bladder outlet resistance. It is possible to estimate the probability to void to completion quite accurately on the basis of bladder outlet resistance and bladder contractility. We named this probability "relative bladder outlet resistance." A high probability of a postvoid residual urine volume may be assumed to indicate "relative BOO." By its very nature, the correlation between "relative BOO" and postvoid residual urine volume is good. PMID- 14550441 TI - Comparison of outcomes of transurethral prostate resection in urodynamicallyobstructed versus selected urodynamicallyunobstructed or equivocal men. AB - OBJECTIVES: To compare the benefits of transurethral resection of the prostate in urodynamically obstructed versus selected urodynamically unobstructed or equivocal men with severe lower urinary tract symptoms associated with benign prostatic enlargement. METHODS: In this case series study, men with lower urinary tract symptoms were selected if they met the study criteria and underwent the tests recommended by the International Scientific Committee on Benign Prostatic Hyperplasia. They also underwent urodynamic investigations. Men were included when transurethral resection of the prostate was selected as the treatment modality. Of the 132 included subjects, 93 could be re-evaluated 6 months after surgery. RESULTS: Of the 93 re-evaluated men, 59 were obstructed and 34 were unobstructed or equivocal. Both groups were similar with respect to age, symptoms, bother, benign prostatic hyperplasia-impact index, and quality of life. The quantified reductions in symptoms and bother in the unobstructed and equivocal men were about 70% of those reductions in the obstructed men. In the equivocal men, and even in the unobstructed men, a significant reduction with 40% of the urethral resistance occurred. CONCLUSIONS: Transurethral resection of the prostate may be a good treatment alternative for unobstructed or equivocal men with severe lower urinary tract symptoms associated with prostatic enlargement, who opt for resection or who do not respond to or do not tolerate medical therapy. PMID- 14550442 TI - Dietary manipulation, ethnicity, and serum PSA levels. AB - OBJECTIVES: To examine whether a diet low in fat and high in fiber, fruits, and vegetables and ethnicity had any influence on serum prostate-specific antigen (PSA) levels, because serum PSA is a marker for the presence of prostate cancer. The incidence of prostate cancer increases with age, varies by ethnicity, and is greater among men with a first-degree relative who has had the disease. Large international variations in the rates of prostate cancer incidence and mortality, as well as the incidence changes in migrants and their offspring, also suggest that exogenous factors, including diet, have a strong influence on the development of this disease. METHODS: We used data and blood samples from the Polyp Prevention Trial, a multicenter randomized trial designed to evaluate the impact of a diet low in fat and high in fiber, fruits, and vegetables on the recurrence of colorectal adenomas. Recruitment was from 1991 through 1994. Participants were followed up from their baseline recruitment date for 4 years. From this group, we identified 1100 white men and 97 black men who were 35 years of age or older, did not have prostate cancer, and had serum samples available for study. RESULTS: At baseline, no difference was present in the fat intake for the black and white men (mean +/- SE, 90 +/- 3.6 g/day and 84 +/- 1.0 g/day, respectively; P = 0.15). The baseline serum PSA levels did not vary by ethnicity. For black men, the mean serum PSA level was 2.2 +/- 0.36 ng/mL compared with 2.0 +/- 0.07 ng/mL for white men (P = 0.64). Although all men assigned to the intervention group markedly reduced their fat intake by approximately 15% and increased their fruit and vegetable intake by approximately 2.25 servings per day, no difference was noted in the kinetics of the serum PSA levels by dietary intervention or race. CONCLUSIONS: Although ethnic differences in the incidence of prostate cancer are well defined, we found no difference in the baseline fat intake among black and white men that might have contributed to this difference. Serum PSA, a marker often used in early detection programs for prostate cancer, was not associated with manipulation of the amount of fat in the diet, regardless of ethnicity. PMID- 14550443 TI - Clinical correlates to PSA spikes and positive repeat biopsies after prostate brachytherapy. AB - OBJECTIVES: To make some preliminary observations regarding the biochemical characteristics of the doubly confusing picture of prostate-specific antigen (PSA) spikes and histologically positive biopsies after prostate brachytherapy. METHODS: All patients reported here had a pretreatment PSA level of less than 10 ng/mL and Gleason score of 4 to 6. Transperineal iodine-125 implants (without supplemental beam radiotherapy) were performed as previously described. After implantation, patients were followed up routinely, with repeat PSA measurements and physical examinations every 4 to 6 months. The timing of the postimplant PSA measurements was at the discretion of the patients and their doctors. No patient received preimplant or postimplant hormonal therapy. Repeat biopsies were performed from 13 to 31 months (median 22) after implant. RESULTS: Patients' prespike nadir ranged from 0.9 to 1.7 ng/mL (median 1.2). The time from the implant to the start of the spike ranged from 9 to 24 months (median 13). The time from implant to the spike peak ranged from 12 to 30 months (median 22). The peak spike height ranged from 2.6 to 8.4 ng/mL (median 3.1). Patients' last PSA value ranged from 0.1 to 0.5 ng/mL (median 0.2). CONCLUSIONS: Transient PSA rises can occur even in the presence of a persistently positive biopsy, and patients and physicians should not feel compelled to rush ahead with salvage therapy. On the basis of the patient data reported here, it appears that a spike up to 10 ng/mL is still consistent with cancer eradication. PMID- 14550445 TI - Prospective evaluation of pain medication requirements and recovery after radical perineal prostatectomy. AB - OBJECTIVES: To perform a study to quantify the variables relating to postoperative pain, activity, and gastrointestinal function after radical perineal prostatectomy to allow comparisons with alternative treatments. METHODS: Ninety-eight consecutive radical perineal prostatectomy candidates between January 2001 and December 2001 with clinically localized prostate cancer were prospectively evaluated. The time to tolerate solid food, time to unassisted ambulation, postoperative pain levels (analog pain scale of 1 to 10), and perioperative analgesic requirements (in morphine equivalents) were selected as the analysis endpoints and correlated with preoperative (age, American Society of Anesthesiology class, body mass index, and serum prostate-specific antigen level), intraoperative (node dissection, operating room time, and estimated blood loss), and postoperative (Gleason score, tumor stage, and lower extremity neurapraxia) patient variables. RESULTS: The mean time to tolerate solid food and unassisted ambulation was 21.2 +/- 1.4 and 22.4 +/- 0.8 hours, respectively; 25.5% of patients experienced transient lower extremity neurapraxia, which was associated with longer operative times (P = 0.001). In a multivariate regression analysis, lymph node dissection correlated with both a prolonged time to tolerate solid food (P = 0.002) and unassisted ambulation (P = 0.001) and neurapraxia with an extended time to unassisted ambulation (P = 0.018). The narcotic requirements were greatest on postoperative day 1, totaling 31.7 +/- 3.0 morphine equivalents, of which 90.5% +/- 3.1% were met with oral analgesics. The average maximal pain scores were highest the first week after discharge (4.7 +/- 0.3), yet approached baseline levels by 4 weeks (1.7 +/- 0.2) after surgery at which time no patient required any pain medication. CONCLUSIONS: Modern radical perineal prostatectomy offers a favorable outcome profile with early patient recovery and low narcotic requirements. A future prospective study should directly compare radical perineal, retropubic, and laparoscopic prostatectomy to document whether the latter offers any advantages with respect to these outcome parameters. PMID- 14550444 TI - Pain assessment after original transperineal prostate biopsy using a coaxial needle. AB - OBJECTIVES: To assess whether the use of a coaxial needle reduces discomfort in patients undergoing multiple-core transperineal prostate biopsy to detect prostate cancer. METHODS: From October 2002 to January 2003, we enrolled 102 consecutive patients with a suspicion of prostate cancer. In every case, we performed a 14-core transperineal prostate biopsy under transrectal ultrasound guidance. The patients were randomized into two groups: group 1 (n = 51) in which we used the 17-gauge coaxial TruGuide needle, and group 2 (n = 51) in which the conventional transperineal technique was used. At the end of the procedure, patients were asked to complete a questionnaire regarding the level of pain experienced. RESULTS: The studied groups were comparable in age, total prostate specific antigen value, and prostate volume. The whole procedure was significantly less painful in group 1 (2.20 +/- 1.20 versus 2.90 +/- 1.73, P = 0.01). We failed to show any significant pain score differences during rectal probe insertion (P = 0.10), transrectal ultrasonography (P = 0.16), and execution of local anesthesia (P = 0.11). The pain score recorded during the multiple-core prostate sampling was significantly lower in group 1 (1.53 +/- 1.5 versus 2.43 +/ 1.86, P = 0.009). No statistically significant differences were found in the complication rates between the two groups. CONCLUSIONS: The use of a coaxial needle reduces the procedure's invasiveness and patient's pain compared with the conventional transperineal prostate biopsy. PMID- 14550446 TI - Comparison of the American Society of Anesthesiologists Physical Status classification with the Charlson score as predictors of survival after radical prostatectomy. AB - OBJECTIVES: To compare the American Society of Anesthesiologists Physical Status (ASA) classification with the Charlson score in the radical prostatectomy setting. The ASA classification is a widely accepted way to evaluate perioperative risk. At present, the Charlson score is probably the most frequently used comorbidity measure to predict long-term survival after radical prostatectomy. METHODS: A total of 444 consecutive patients were enrolled in this study. The ASA classification was obtained from the anesthesia chart, and the Charlson score was assigned based on conditions noted during the preoperative cardiopulmonary risk assessment or mentioned on the discharge document. Kaplan Meier time-event curves and Mantel-Haenszel hazard ratios were estimated for comorbid (noncancer) and overall survival. RESULTS: After a mean follow-up of 5.9 years, both classifications were able to predict comorbid and overall survival in dose-response patterns. The ASA classification was superior in terms of a clearer discrimination of the survival curves (lower P values, higher hazard ratios). Both classifications identified a high-risk group (ASA 3 and Charlson score 2 or more), but only the ASA classification sufficiently defined a low-risk group (ASA 1). CONCLUSIONS: In experienced hands, the ASA classification is a promising tool to improve the classification of prognostic comorbidity in the radical prostatectomy setting and may be used as an alternative to the Charlson score. PMID- 14550447 TI - Contemporary evaluation of operative parameters and complications related to open radical retropubic prostatectomy. AB - OBJECTIVES: To determine the impact of recent advances in surgical technique, management, and early detection on outcome after open radical retropubic prostatectomy. METHODS: Between October 2000 and August 2002, 500 men with clinically localized prostate cancer underwent radical retropubic prostatectomy by a single surgeon (H.L.). One of the unique aspects of this prospective outcomes analysis was that both data acquisition and entry were conducted totally independent of the primary surgeon. RESULTS: The mean operative and prostatectomy time was 142.9 and 65.2 minutes, respectively. A single ureteral injury was the only intraoperative complication. The overall incidence of pulmonary embolus and/or deep vein thrombosis was 0.4%. The overall risk of allogeneic transfusion was 4.6%. The mean length of hospital stay was 2.11 +/- 0.04 days. Of the catheters, 83.6% were removed by postoperative day 8. The positive surgical margin rate was 8%. CONCLUSIONS: In the hands of experienced surgeons, outcomes after open radical prostatectomy are excellent. Laparoscopic and robotic prostatectomy must be compared with concurrent experiences with open radical prostatectomy. PMID- 14550448 TI - Quality-of-life questionnaire results 2 and 3 years after radiotherapy for prostate cancer in a randomized dose-escalation study. AB - OBJECTIVES: To assess patient-reported prostate cancer-specific quality of life 2 and 3 years after radiotherapy to the prostate in a randomized dose-escalation trial of 70 versus 78 Gy conducted from 1993 to 1998. METHODS: Two years after completing radiotherapy, a questionnaire that assessed bladder, rectal, and sexual function was sent to 301 patients in the study. Three years after treatment, a second questionnaire was sent to the 175 patients with adequate follow-up. RESULTS: Three years after radiotherapy, urinary incontinence was reported by 35% of patients, but only 6% required the use of a pad or other protective device. Patients reported increased leakage with a full bladder (urge incontinence) between the 2 and 3-year questionnaires (42% versus 50%; P = 0.03). At 3 years, 33% of patients reported rectal bleeding compared with 47% at 2 years (P = 0.006). Patients in the 78-Gy arm reported more frequent bowel movements at 3 years and less change in bowel function at 2 years than patients in the 70-Gy arm. Before radiotherapy, 84% of patients reported erections adequate for intercourse at least a few times during the previous year. After 2 and 3 years, this had decreased to 49% and 41%, respectively (P <0.02). CONCLUSIONS: By patient-reported questionnaire, 78 Gy produced an increase in bowel movement frequency and no increase in bladder or sexual side effects at 3 years compared with 70 Gy. Comparing the results 2 and 3 years after radiotherapy, the symptoms of rectal bleeding had improved, erectile function had decreased, and urinary urge incontinence had increased. PMID- 14550449 TI - Simple sling incision for the treatment of iatrogenic urethral obstruction. AB - OBJECTIVES: To review a single-institution experience with sling incision to treat urethral obstruction resulting from pubovaginal sling placement. Urethrolysis involving full mobilization of the urethra has traditionally been used to treat urethral obstruction resulting from a pubovaginal sling. Recently, sling incision has been reported to be a technically simpler, less potentially morbid technique to relieve such obstruction. METHODS: The charts of 14 women with iatrogenic urethral obstruction from pubovaginal slings who were treated with simple sling incision were reviewed. Presenting symptoms and physical findings and outcomes (primarily resolution of obstructive and irritative symptoms, change in postvoid residual volume, and change in continence status) were evaluated. RESULTS: Eleven patients had a biologic sling and three a mid urethral polypropylene mesh sling. All patients had either obstructive or irritative findings or retention and 10 of 14 had an elevated postvoid residual volume. The average follow-up was 12 months. Overall, 13 (93%) of 14 had complete or significant improvement of presenting complaints, and 1 (7%) eventually required formal urethrolysis. Of the 13 with resolution of presenting complaints, 3 had residual irritative symptoms. Although 3 (21%) of 14 had some element of recurrent stress urinary incontinence, only 1 (7%) required treatment. CONCLUSIONS: Simple sling incision is an effective, technically simple, and safe treatment for women with iatrogenic urethral obstruction after placement of a pubovaginal sling. Its results are comparable to those of formal urethrolysis, and it may be considered as first-line treatment for iatrogenic obstruction after a sling procedure. PMID- 14550450 TI - Failed posterior urethroplasty: lessons learned. AB - OBJECTIVES: To determine the factors contributing to unsuccessful results after posterior urethroplasty and to establish some guidelines for its prevention. METHODS: From 1977 through 2000, 130 patients (3 to 58 years old) underwent 145 anastomotic urethroplasty procedures for post-traumatic posterior urethral distraction defects. Bulbo-prostatic anastomosis was performed through the perineum in 105 cases and by a perineo-abdominal transpubic procedure in 40. Factors that might have an impact on the surgical result, such as the size and type of sutures, urethral stents, and use of antibiotics were standardized. The records of all patients with unsuccessful results during a follow-up of 2 to 20 years were analyzed. RESULTS: Overall, the results were unsuccessful in 12 cases (8%). A negative result was related to incomplete excision of the scar tissue in 3 cases, inadequate fixation of the prostatic mucosa in 6 cases, and anastomotic tension in 2 cases; 1 patient was lost to follow-up. Previous repair, length of distraction defect, and urinary infection had no statistically significant influence on the result. CONCLUSIONS: The essential operative details of posterior urethroplasty include complete excision of scar tissue involving the membrano-prostatic region, lateral fixation of pliable prostatic mucosa, and creation of a tension-free anastomosis. If a tension-free anastomosis cannot be achieved through the perineum, the perineo-abdominal progressive approach or the elaborated perineal technique should be used at the same setting. Previous repair, a long distraction defect, and urinary infection do not preclude successful posterior urethroplasty. PMID- 14550451 TI - Comparison of outcomes after single or DOUBLE-CUFF artificial urinary sphincter insertion. AB - OBJECTIVES: To assess the effectiveness and complications associated with single and double-cuff artificial urinary sphincter (AUS) implantation for postprostatectomy stress urinary incontinence. METHODS: A retrospective study of 56 men with postprostatectomy stress urinary incontinence who underwent either single (28 patients) or double (28 patients) cuff AUS placement was performed. Patients in each cohort were matched on the basis of preoperative pad use, risk factors for complications, and age. Patient selection was blinded relative to outcome. Continence, quality of life, and complications were assessed using the Incontinence Impact Questionnaire Short Form (IIQ-7), postoperative pad use, and chart review. RESULTS: The mean age was 67 years for each group. Daily pad use decreased from 7.7 to 1.1 in patients treated with a single-cuff AUS and from 7.8 to 0.7 in patients with a double-cuff AUS (P = 0.25). Complete continence (0 pads daily) was reported in 3 (11%) of 28 men with single-cuff and 12 (43%) of 28 men with double-cuff sphincters (P = 0.008). The IIQ-7 scores improved from 14.8 to 3.1 after single-cuff placement and from 16.3 to 2.5 after double-cuff placement (P = 0.03). With an average follow-up of 41.3 and 21.2 months for the single and double-cuff cohorts, respectively, five complications were reported in the single cuff recipients and four in the double-cuff patients. CONCLUSIONS: A significantly greater rate of complete continence and improvement in the IIQ-7 were seen in men with double-cuff AUS compared with single-cuff devices. Additional study is needed to confirm the relative advantages of double-cuff insertion. PMID- 14550452 TI - Biochemical screening in the assessment of erectile dysfunction: what tests decide future therapy? AB - OBJECTIVES: To evaluate which biochemical tests, performed as screening tests in the assessment of erectile dysfunction (ED), lead to a change in standard treatment. METHODS: We examined the results of the biochemical and endocrine tests performed as part of the assessment of 1455 men presenting with ED. We also documented the subsequent therapy. RESULTS: We found abnormal findings in testosterone (5.7%), prolactin (0.5%), thyroid function tests (0.13%), liver function tests (12.8%), glucose (9.3%), cholesterol (15% in a subset of 531 men), and ferritin (1.6%). Hyperprolactinemia was not invariably associated with a suppressed testosterone level. Specific therapy with a dopamine agonist for hyperprolactinemia and with testosterone for hypogonadism was effective in treating ED. For the others, standard ED therapy was used. CONCLUSIONS: The results of our study showed that screening tests fall into two categories. Testosterone and prolactin have a low yield but specific therapy is effective. Glucose and lipids have a higher yield but specific therapy is not immediately effective for ED. However, they, on their own merits, require management to prevent long-term morbidity and mortality. PMID- 14550453 TI - Reoperative retroperitoneal surgery for nonseminomatous germ cell tumor: clinical presentation, patterns of recurrence, and outcome. AB - OBJECTIVES: To describe the clinical characteristics and outcome of patients with metastatic nonseminomatous germ cell tumor requiring reoperative retroperitoneal surgery at the Memorial Sloan-Kettering Cancer Center, because such patients are poorly characterized. METHODS: The Memorial Sloan-Kettering Cancer Center germ cell tumor surgical database was reviewed from January 1989 through April 2001, and the clinical characteristics of patients undergoing reoperative retroperitoneal surgery for nonseminomatous germ cell tumor were identified. The initial presentation, histologic findings, morbidity, and survival were analyzed. Disease-specific survival was calculated using the Kaplan-Meier method. RESULTS: A total of 56 patients underwent 61 repeat operations: 22 after primary retroperitoneal lymph node dissection (RPLND) and 34 after postchemotherapy RPLND. Left testicular primary tumors were more common than right (33 versus 23), and the most common sites of disease prompting reoperation were the para-aortic and left hilar regions. Teratoma was the most common histologic finding at the time of reoperation. Of 56 patients, 37 (66%) required chemotherapy between the initial operation and reoperation. The overall perioperative complication rate was 27%, and median length of hospital stay was 8 days. Sixty-nine percent of patients required adjunctive procedures at the time of reoperation, the most common of which was thoracotomy. The 5-year disease-specific survival rate was 67% for the entire group (86% following reoperation after primary RPLND and 56% following reoperation after postchemotherapy RPLND). CONCLUSIONS: Reoperative retroperitoneal surgery for nonseminomatous germ cell tumor can be performed with acceptable morbidity in select referral centers. Teratoma is highly prevalent in the retroperitoneum at the time of reoperation. A significant subset of these high-risk patients can be salvaged with complete resection. PMID- 14550454 TI - Risk factors for spontaneous bladder perforation after augmentation cystoplasty. AB - OBJECTIVES: To present our experience with the use of stomach, ileum, and colon for augmentation cystoplasty to examine the incidence of, and risk factors for, spontaneous perforation. Spontaneous bladder perforation is a potentially life threatening complication of augmentation cystoplasty with a reported incidence of up to 13%. METHODS: A retrospective review of medical records from 1988 to 2001 identified 107 children (57 males and 50 females) who underwent augmentation cystoplasty at our institution. The etiology for bladder dysfunction included myelomeningocele, VATER (vertebral defects, imperforate anus, tracheoesophageal fistula, radial and renal dysplasia) syndrome, bladder and cloacal exstrophy, posterior urethral valves, and pelvic malignancy. Thirteen patients also had end stage renal disease. RESULTS: The median follow-up was 7.4 years. Gastrocystoplasty was performed in 50 children (47%), ileocystoplasty in 37 (35%), colocystoplasty in 18 (17%), and gastric-ileal composite neobladder in 2 (2%). Augmentation cystoplasty procedures were performed using a standard technique that included a two-layer anastomosis. Additional procedures at the time of reconstruction included Mitrofanoff neourethra in 66 patients (62%) and bladder neck repair in 44 (41%). Postoperatively, most patients started a strict incremental catheterization regimen. The overall incidence of bladder perforation was 5%, with one traumatic (1%) and four spontaneous (4%) perforations. All patients recovered uneventfully after exploratory laparotomy. CONCLUSIONS: We believe that the relatively low incidence of spontaneous bladder perforation encountered in this series may be explained by the large number of patients with gastrocystoplasty, as well as our strict adherence to a postoperative incremental catheterization program. PMID- 14550455 TI - Adolescent varicocele: is it a unilateral disease? AB - OBJECTIVES: To evaluate the incidence of left and right varicocele in adolescents. METHODS: The study group consisted of 28 adolescents who underwent evaluation for varicocele at our clinic. In 19 patients, varicocele was detected on routine physical examination, and 9 patients presented with scrotal pain or discomfort. All patients were evaluated by three methods. Physical examination was followed in all cases by contact thermography and venography studies of both testes. RESULTS: The rate of left and right retrograde flow in the spermatic veins by the three methods used was as follows: physical examination 92.8% and 10.7%; contact thermography 100% and 89%; and venography 100% and 85.7%, respectively. Varicocele was not detected by physical examination in 2 (7.2%) of the 28 patients on the left side and in 21 (87.5%) of 24 on the right side. CONCLUSIONS: The main finding of this study was that varicocele is a bilateral disease in 85.7% of adolescents. The high percentage of bilateral varicocele in our sample may explain the pathophysiologic mechanism whereby what has traditionally been considered a unilateral disease can produce bilateral testicular dysfunction. The high incidence of subclinical bilateral varicocele may indicate that we should consider varicocele a bilateral disease. The second finding was that only 10% of patients with right varicocele were diagnosed by physical examination and more than 85% were diagnosed by thermography, with confirmation by venography. Therefore, we suggest that thermography and venography should play a major role in the diagnosis of varicocele. PMID- 14550458 TI - Transitional cell carcinoma in a fused crossed ectopic kidney. AB - Nephroureterectomy is considered the reference standard treatment for invasive transitional cell carcinoma of the ureter. When this malignancy occurs in a patient with renal fusion anomaly, separating the involved kidney from its conjoint mate becomes a challenging task for the surgeon. We report a unique case in which a patient with an L-shaped left-to-right crossed ectopic kidney was diagnosed with invasive ureteral transitional cell carcinoma. The preoperative assessment in these uncommon cases should include renal angiography or computed tomography angiography to provide key information about the renal vasculature, which is essential for planning the surgical dissection and line of separation between the kidneys. PMID- 14550459 TI - Distinguishing splenosis from renal masses using ferumoxide-enhanced magnetic resonance imaging. AB - A 43-year-old man with a history of splenectomy was found to have a solid renal mass on computed tomography. Magnetic resonance imaging with ferumoxide characterized this mass as ectopic splenic tissue and nephrectomy was avoided. PMID- 14550461 TI - Extensive palliative surgery for advanced mesothelioma of the tunica vaginalis. AB - Malignant mesothelioma of the tunica vaginalis is a rare tumor managed principally by radical surgical resection. Chemotherapy and radiotherapy have limited efficacy. We report on a 67-year-old man with severe debilitation from multiple scrotal and inguinal recurrences of a malignant mesothelioma originating in the right tunica vaginalis. Local pain from extensive tumor spread prevented ambulation. Aggressive surgical debridement (total penectomy and scrotectomy) and perineal urethrostomy afforded the patient significant improvement in his quality of life before he finally died of the disease 3 years after diagnosis. PMID- 14550460 TI - Prostate carcinoma presenting as multiple pulmonary nodules in an asymptomatic patient with a history of testicular nonseminomatous germ cell tumor. AB - Prostate carcinoma presenting initially as multiple pulmonary nodules in an asymptomatic patient without previous prostate carcinoma is unusual. Whether the incidence of prostate carcinoma is significantly increased in patients treated previously for germ cell tumors is unclear. We report such a patient, who responded to combination androgen blockade therapy. PMID- 14550462 TI - Obstructive nephropathy secondary to sulfasalazine calculi. AB - The incidence of drug-induced stone disease is 0.44%. A 57-year-old woman with ulcerative colitis presented with obstructive nephropathy and pyelonephritis. She underwent cystoscopy, bilateral retrograde pyelography, and bilateral ureteral stent placement. A 6-cm bladder calculus and two 3-mm right distal ureteral calculi were discovered. Later, cystolithotomy was performed. The stone analysis demonstrated sulfapyridine, a sulfasalazine metabolite. Patients with inflammatory bowel disease can develop urolithiasis owing to acidic urine and low volume urine production. Patients receiving aminosalicylates are at an increased risk of urolithiasis and may benefit from oral hydration and urinary alkalization. PMID- 14550463 TI - Two secondary malignancies after radiotherapy for seminoma: case report and review of the literature. AB - We report a case of a 50-year-old man with two synchronous second malignancies 25 years after orchiectomy and adjuvant radiotherapy for seminoma. An annual health examination revealed an elevated prostate-specific antigen level. A biopsy was performed revealing Gleason score 9 adenocarcinoma of the prostate. Computed tomography of the abdomen revealed a 2-cm solid mass in the right kidney consistent with renal cell carcinoma. Both of these lesions were within the nonstandard radiation field for seminoma with which this patient was treated. Second malignancies, including prostate cancer, are a very uncommon occurrence but an important consideration in long-term survivors of seminoma treated with radiotherapy. PMID- 14550464 TI - Renal-ovarian axis: a case report and review. AB - The renal-ovarian axis is a unique and rare phenomenon manifested by an interesting interrelationship between the ovary and kidney. The similar embryologic development, the histologic similarities of these malignancies, and asymmetric venous outflow represent the foundation of this theoretical axis. A poor understanding of this metastatic spread has led to missed, or delayed, diagnosis of the primary malignancy. We describe the only case of a late recurrent metastatic ovarian carcinoma masquerading as a solitary renal cell carcinoma. An exhaustive review of published English reports evaluating the renal ovarian axis was undertaken to characterize and understand the nuances of this entity. PMID- 14550465 TI - Large cerebellar lesion as original manifestation of transitional cell carcinoma of the bladder. AB - Patients with transitional cell carcinoma of the bladder classically present with irritative voiding symptoms or painless hematuria. Common sites of vascular metastases include the liver (38%), lung (36%), bone (27%), adrenal glands (21%), and intestine (13%). Vascular metastasis to the brain, without a prior history of chemotherapy, is extremely rare. To our knowledge, this is the first report of a patient with bladder transitional cell carcinoma whose original presentation was from a symptomatic, metastatic, cerebellar lesion. PMID- 14550466 TI - Rapamycin preserves renal function compared with cyclosporine A after ischemia/reperfusion injury. AB - OBJECTIVES: To determine the effect of cyclosporine and rapamycin administration on renal function after ischemia/reperfusion injury (I/R). Cyclosporine A has known nephrotoxic effects. Thus, cyclosporine therapy subsequent to I/R injury may further exacerbate graft dysfunction. Rapamycin is a newer agent that suppresses the immune system by a different mechanism. METHODS: Male Wistar rats (250 g) were anesthetized, and the suprarenal aorta was clamped for 40 minutes. The right kidney was removed. After recovery, the rats were divided into four groups: group 1, controls, no ischemia and no treatment (n = 10); group 2, ischemia with no treatment (n = 8); group 3, ischemia plus rapamycin (0.17 mg/kg/day gavage, n = 8); and group 4, ischemia plus cyclosporine A (30 mg/kg/day intraperitoneally, n = 9). The glomerular filtration rate was measured 5 to 7 days after I/R injury using urinary iohexol clearance. Data are expressed as the mean +/- SEM, and intergroup comparisons were made using one-way analysis of variance. RESULTS: The mean GFR value for the controls (no ischemia, no treatment) was 1.23 +/- 0.08 mL/min; for group 2 (ischemia, no treatment), it was 1.05 +/- 0.10 mL/min; for group 3 (ischemia plus rapamycin) 1.06 +/- 0.14 mL/min; and for group 4 (ischemia plus cyclosporine A) 0.44 +/- 0.06 mL/min (P <0.05 versus the other three groups). The mean arterial pressure was significantly lower in the ischemic rats treated with cyclosporine A (P <0.05 versus the other three groups). CONCLUSIONS: After I/R injury, rapamycin may preserve renal function compared with cyclosporine treatment, because it does not have a direct vasoconstrictor effect on the renal microcirculation. PMID- 14550467 TI - Effect of spinal cord injury on urinary bladder spinal neural pathway: a retrograde transneuronal tracing study with pseudorabies virus. AB - OBJECTIVES: To determinate the effect of acute and chronic spinal cord injury (SCI) resulting from thoracic cord transection on the urinary bladder spinal neural pathway. METHODS: Seventy-six adult Sprague-Dawley rats were randomly divided into four groups, non-SCI (normal rats undergoing no surgical procedure except pseudorabies virus [PRV] injection), SCI(b) (SCI and PRV injected immediately after SCI), SCI(c) (SCI and PRV injected at 3 weeks after SCI), and SCI(d) (SCI and PRV injected at 3 months after SCI). Transcardiac perfusion fixation was done at appropriate survival periods after PRV injection into the bladder wall tissue. Sections of the dorsal root ganglion, spinal cord, and brain were processed for visualization of the virus by the streptavidin-peroxidase immunohistochemical procedure. RESULTS: The bladder weight of the non-SCI, SCI(b), SCI(c), SCI(d) rats was 144 +/- 9 mg, 142 +/- 8 mg, 486 +/- 51 mg, and 656 +/- 69 mg, respectively. The time-ordered flow charts of PRV tracing were similar in the non-SCI and SCI rats. The cross-sectional area of the labeled dorsal root ganglion cell profiles increased significantly after SCI (P <0.001): 593 +/- 40 microm2, 588 +/- 39 microm2, 815 +/- 53 microm2, and 902 +/- 57 microm2 in the non-SCI, SCI(b), SCI(c), SCI(d) rats, respectively. The number of labeled cells in the dorsal horn in the L6 and S1 segments 3 days after PRV injection markedly increased in chronic SCI rats, as did the number of labeled motor neurons 4 days after injection. CONCLUSIONS: Acute and chronic SCI have no effect on the process of virus transneuronal transport below the level of the lesion. Subsequent to chronic SCI, reorganization of the micturition reflex pathways may occur. PMID- 14550468 TI - Reperfusion of ischemic corporal tissue: physiologic and biochemical changes in an animal model of ischemic priapism. AB - OBJECTIVES: To assess the physiologic and biochemical changes resulting from ischemia and reperfusion. Effective therapy for ischemic priapism reestablishes corporal venous outflow and arterial inflow and results in increased corporal partial pressure of oxygen. Data are limited concerning reperfusion injury of ischemic erectile tissue associated with reactive oxygen species (ROS) and the potential role of ROS scavengers in the clinical therapy of ischemic priapism. METHODS: Anesthetized adult New Zealand white male rabbits (n = 7) were exposed to a low oxygen tension breathing gas to achieve hypoxia within the corpora cavernosa. This resulted in a mean systemic oxygen saturation of 60%. The pelvic nerve was electrically stimulated to induce penile erection, and the base of the erect penis was clamped. After varying durations of ischemia, the clamp was removed to allow reperfusion. We determined the intracavernosal oxygen tension, histologic changes, myeloperoxidase activity, and lipid peroxidation. RESULTS: Corporal partial pressure of oxygen progressively decreased as the duration of priapism increased. A statistically significant increase was noted in myeloperoxidase activity and lipid peroxidation with corporal reperfusion. Polymorphonuclear leukocyte infiltration was documented in the ischemic reperfused tissue. CONCLUSIONS: In the management of ischemic priapism, reperfusion causes erectile tissue injury owing to the presence of ROS. There is a need to investigate the utility of ROS scavengers and antioxidants in the management of ischemic priapism. PMID- 14550469 TI - Immature tumor angiogenesis in high-grade and high-stage renal cell carcinoma. AB - OBJECTIVES: To investigate the correlation between pathologic findings and maturation of the tumor neovasculature of renal cell carcinoma by immunohistochemical studies. METHODS: Formalin-fixed and paraffin-embedded specimens from 25 randomly selected patients with renal cell carcinoma were stained with mouse monoclonal antibodies, anti-human CD31, anti-alpha smooth muscle actin (alphaSMA), and anti-human calponin by the indirect immunoperoxidase method. The microvessels were counted in six areas with the higher number of microvessels in each patient at 200x magnification (0.255 mm2 per area). RESULTS: The number of CD31-positive microvessels in grade 3 tumors was significantly lower than those in grade 1 or 2 tumors (P = 0.003222 and P = 0.043217, respectively). The CD31-positive microvessel counts of those of higher stage, tumor size greater than 4.5 cm, or non-clear cell type were significantly lower than tumors of lower stage, size less than 4.6 cm, or clear cell type. In the grade 3 tumors, the expression ratio of the number of alphaSMA-positive microvessels to the number of CD31-positive microvessels was significantly decreased compared with grade 1 or 2 tumors (P = 0.000011 and P = 0.000000, respectively). The expression of calponin in the tumor neovasculature was not observed. The expression ratios of the number of alphaSMA-positive microvessels to the number of CD31-positive microvessels in higher stages, larger tumor sizes, or non-clear cell types were significantly decreased. CONCLUSIONS: The tumor neovasculature of high-grade and high-stage tumors was immature. These results imply that high-grade tumors of renal cell carcinomas may be susceptible to antiangiogenesis therapy inducing apoptosis of immature tumor vessels. PMID- 14550470 TI - Quantitative RT-PCR assay for MUC3 and VEGF mRNA in renal clear cell carcinoma: relationship with nuclear grade and prognosis. AB - OBJECTIVES: To investigate, by real-time quantitative reverse transcriptase polymerase chain reaction, the expression of MUC3 and vascular endothelial growth factor (VEGF) to correlate them with histologic parameters and with prognosis. Human mucins are large O-glycoproteins expressed in epithelial tissues. Deregulation of mucin genes has been demonstrated in several epithelial neoplasms. In the kidney, MUC3 is expressed in normal convoluted tubules and in renal clear cell carcinoma. METHODS: Twenty-six renal clear cell carcinoma specimens were studied. For all tumors, samples of normal and tumor kidney were frozen. After RNA extraction, using ultracentrifugation through a cesium chloride cushion, VEGF and MUC3 mRNA were analyzed by real-time quantitative reverse transcriptase-polymerase chain reaction. The pathologic parameters included Fuhrman nuclear grade and TNM stage. All follow-up data were available. RESULTS: The median level of MUC3 and VEGF expression was greater in tumor areas compared with normal areas (P < 0.002 and P < 0.001, respectively). The MUC3 tumor/normal tissue expression ratio was greater in nuclear grade 3 tumor than in low grades (grade 1-2; P < 0.005). No statistically significant relationship was found with the prognosis for MUC3 and VEGF in our study. CONCLUSIONS: The results of this study demonstrate that MUC3 and VEGF are overexpressed in renal clear cell carcinoma, and the MUC3 expression ratio is greater in nuclear grade 3 than in grades 1 and 2 (low grades) tumor. These findings suggest the implication of MUC3 in renal carcinogenesis. PMID- 14550472 TI - Shortened pull-string simplifies office-based ureteral stent removal. PMID- 14550471 TI - Urologic techniques of Serefeddin Sabuncuoglu in the 15th century Ottoman period. PMID- 14550474 TI - Effective human defense against E. histolytica: high amoebicidal activity of lymphocytes and monocytes in amoebic liver abscess patients until 3 months follow up. AB - Adherence of pathogenic Entamoeba histolytica trophozoites mediated by Gal/GalNAc lectin is a prerequisite for killing naive T cells and monocytes but the activated T cells and monocyte derived macrophages (MDMs) not only resist the attack but can kill the parasite. In the present study, we have analysed the adherence and cytotoxicity of the immunecompetent cells from patients of amoebic liver abscess at the time of their diagnosis and after 3 months to elucidate the development of cell mediated cytotoxicity, a major mechanism of resistance to amoebic infection. The results show that CD3+ cells from amoebic liver abscess cases, when stimulated, in vitro, bound E. histolytica trophozoites with increased intensity and their viability was also increased. The activated lymphocytes (taken at 3 months post treatment) were also able to kill amoebae. MDMs bound amoebae with greater intensity than lymphocytes, until 3 months post infection. These MDMs were effective in killing approximately 40% amoebae which was significantly less than at the time of diagnosis but was very significant as compared to the controls. The data suggest that cell mediated cytotoxic responses are maximum until 1 month post treatment and are significantly reduced thereafter. PMID- 14550476 TI - Effects of condensed tannins and crude sesquiterpene lactones extracted from chicory on the motility of larvae of deer lungworm and gastrointestinal nematodes. AB - The objective of this study was to determine the effects of condensed tannins (CT) and an extract containing crude sesquiterpene lactones (CSL) from chicory (Cichorium intybus) on the motility of the first-(L1) and third-stage (L3) larvae of deer lungworm Dictyocaulus viviparus and the L3 larvae of gastrointestinal nematodes in vitro, using the larval migration inhibition (LMI) assay. The CT and CSL had a profound effect on the motility of the larvae displayed by their ability to inhibit larval passage through nylon mesh sieves. Incubation of lungworm L1 larvae in rumen fluid (collected from deer fed pasture) containing 100, 400 and 1000 microg CT/ml, inhibited 12, 28 and 41% of the larvae from passing through the sieves, respectively, while the incubation of L3 larvae with rumen fluid (pH 6.6) containing the same concentrations inhibited 26, 37 and 67% of L3 larvae from passing through the sieves, respectively. Gastrointestinal larvae seem more susceptible to CT than lungworm larvae especially at higher concentrations. CT inhibited 27, 56 and 73% of gastrointestinal larvae from passing through the sieves when used at a concentration of 100, 400 and 1000 microg/ml, respectively. CT were more effective (P<0.001) at reducing the motility of lungworm L1 and L3 larvae when added to the rumen fluid than when added to the abomasal fluid (pH 3.0). Addition of 2 microg polyethylene glycol/microg CT eliminated the inhibitory effect of CT against L1 and L3 larvae especially during incubation in rumen fluid, confirming the effect as due to CT. The CSL extract also showed similar inhibitory activity against L1 and L3 lungworm and L3 gastrointestinal larvae in both fluids, indicating that this extract was not affected by the pH of the fluid, and was more effective against L3 than L1 lungworm larvae. Condensed tannins appeared to be more effective than CSL at inactivating L1 and L3 lungworm and L3 gastrointestinal larvae in rumen fluid, but CSL were particularly effective against L3 lungworm larvae in abomasal fluid. Activity of these secondary compounds explains the reduced parasite problem of young deer grazing chicory. PMID- 14550475 TI - Double aldehyde stabilisation of erythrocytes in the indirect hemagglutination for echinococcosis. AB - Sheep erythrocytes were stabilised with glutaraldehyde tanned and fixed with formalin in the indirect hemagglutination test (IHA-GF) and sensitised with hydatid antigen for the diagnosis of human cystic echinococcosis (CE). The sensitivity of this method was compared to that prepared with fresh tanned cells (IHA-TA) in 278 sera from hydatid patients. The sensitivity of IHA-GF (87.8%) was higher than that of IHA-TA (85.6%), the difference being insignificant. Higher geometric mean titres were obtained by IHA-GF (1:13300) than by IHA-TA (1:11600). The use of two sorts of aldehydes proved to be a satisfactory method, showing high sensitivity, a very good specificity and some advantages. The sensitised cells retained their diagnostic effectiveness for at least 15-18 months when stored at 4 degrees C. The technique is inexpensive and rapid, allowing the testing of a large number of sera. The method reduces the variation of the results and improves the reproducibility of the test. When the minimal diagnostic titre-1:400 is used the specificity of IHA-GF might increase by 2.9% while the sensitivity might decline by only 1.4%. The IHA-GF demonstrated better immunodiagnostic characteristics than enzyme linked immunosorbent assay (ELISA) and Latex agglutination test (LAT). The IHA-GF should be considered as an useful method in the range of classical diagnosis for the serology of CE. The clinical diagnostic potential should be increased by a combination of at least two tests: IHA-GF and ELISA or LAT. PMID- 14550477 TI - Response to re-infection with Brachylaima cribbi in immunocompetent and immunodeficient mice. AB - The course of infection in C57BL/6J mice re-infected with Brachylaima cribbi was assessed by comparing faecal egg excretion of re-infected mice with age- and sex matched mice receiving a primary infection only. For both male and female mice there was a significant reduction in the mean number of eggs per gram of faeces at the peak of infection 4 weeks after the challenge infection compared with mice receiving a primary infection only. There was no significant difference in the duration of the infection. This experiment was repeated using age-matched male mice but on this occasion all mice were killed and dissected 4 weeks after the challenge infection and mean eggs per gram of faeces, worm burden and fecundity determined. There was no significant difference in the worm burdens of the re infected mice compared with age-matched animals receiving a primary infection only. However, there were significant differences in the mean faecal eggs per gram and worm fecundity with the challenge infection group having lower egg counts and reduced fecundity. An enzyme-linked immunosorbent assay using whole worm antigens was developed and used to determine mouse anti-B. cribbi serum antibody levels during the course of infection. Anti-B. cribbi serum antibody absorbance ratios increased six- to sevenfold by 4 weeks after a primary infection beyond which a constant level was maintained. The course of challenge infection in non-obese diabetic severe combined immunodeficient mice showed no significant differences in egg excretion, worm burden or fecundity when primary and challenge infections were compared. These results indicate that the immune response invoked by a previous B. cribbi infection in immunocompetent mice affects fecundity but does not affect the establishment or duration of infection. PMID- 14550478 TI - A multiplex polymerase chain reaction for a differential diagnosis of Plasmodium falciparum and Plasmodium vivax. AB - A multiplex PCR was designed for the differential diagnosis of the two parasite species by targeting the 18S rRNA gene with a set of primer combinations, amplifying DNA fragments of 1451-bp and 833-bp for P. falciparum and P. vivax, respectively. The sensitivity of this PCR test was high, as minimal as 0.1 parasite per one microliter of blood sample and a minimum of four copies of the target gene could be detected. For the diagnosis of mixed infection of two Plasmodium spp., there were no apparent competition or cross-reaction between the majority and minority Plasmodium species. The multiplex PCR was evaluated on 210 clinical samples and 60 normal controls. The PCR test yielded highly concordant results with microscopic examination, with the only one exception of a mixed (P. falciparum plus P. vivax) infection case, which was diagnosed as a single infection of P. falciparum by microscopy. We propose that the multiplex PCR is a sensitive, specific, and rapid tool that can serve as a useful differential diagnostic tool for detecting P. falciparum and P. vivax. PMID- 14550479 TI - Overproduction of highly active trypanosome alternative oxidase in Escherichia coli heme-deficient mutant. AB - Cyanide-insensitive trypanosome alternative oxidase (TAO) is the terminal oxidase of the respiratory chain of long slender bloodstream forms of the African trypanosome, which causes sleeping sickness in humans and nagana in cattle. TAO has been targeted for the development of anti-trypanosomal drugs, because it does not exist in the host. In this study, we established a system for overproduction of highly active TAO in Eschericia coli heme-deficient mutant. Kinetic analysis of recombinant enzyme and TAO in Trypanosoma brucei brucei mitochondria revealed that recombinant TAO retains the properties of native enzyme, indicating that recombinant TAO is quite valuable for further biochemical study of TAO. PMID- 14550480 TI - The largest subunit of the RNA polymerase II of Trypanosoma cruzi lacks the repeats in the carboxy-terminal domain and is encoded by several genes. AB - The largest subunit of eukaryotic RNA polymerase II (RNA Pol II) of several members of Kinetoplastida order and other early diverging eukaryote contains a serine, tyrosine and proline-rich domain in the carboxyl-terminal instead of the typical heptapeptides repeats found in most eukaryotes. The lack of these repeats seems to reflect the different control of gene transcription found in these organisms. To provide further understanding in these mechanisms, we have characterized the largest subunit of RNA polymerase II (RNA Pol II LSU) in Trypanosoma cruzi. We found that it also lacks the heptapeptide repeats in the carboxy-terminal domain, but is quite similar to the same region in the Trypanosoma brucei enzyme, suggesting a conserved role for this domain. In addition, we found several genes encoding the RNA Pol II in T. cruzi, distributed in different chromosomal bands in several isolates. Two of these genes were entirely sequenced and shown to be quite similar. The presence of several gene copies of the RNA Pol II LSU, also described in African trypanosomes might reflect a gene amplification requirement that appeared early in the evolution of these organisms. PMID- 14550481 TI - Helminths of two anuran species, Atelopus spurrelli (Bufonidae) and Dendrobates histrionicus (Dendrobatidae), from Colombia, South America. AB - Two species of anurans from Colombia, South America, Atelopus spurrelli and Dendrobates histrionicus, were examined for helminths. A. spurrelli was found to harbor three species of Nematoda, adults of Cosmocerca podicipinus and larvae of Physocephalus sp. and Porrocaecum sp. D. histrionicus was found to harbor, in addition to C. podicipinus, Physocephalus sp. and Porrocaecum sp. and cystacanths of three species of Acanthocephala: Centrorhynchus sp., Onicola sp. and Polymorphus sp. A. spurrelli and D. histrionicus represent new host records for these helminths. PMID- 14550482 TI - After the viewpoint debate: where next in object recognition? AB - A debate on whether object representations capture isolated viewpoints or ranges of views has dominated research in object recognition in recent years, but now seems to be waning. Rather than assume a narrow model in which either structural or view information is used to recognize an object, researchers have begun to examine how these properties might be used cooperatively. A recent paper by Foster and Gilson confirms sensitivity to both types of information, which combine in an additive framework to predict recognition performance. PMID- 14550483 TI - How does the hippocampus contribute to memory? AB - Recently, Wirth et al. reported that hippocampal neurons signal the acquisition of new associations by altering the selectivity of their responses to crucial stimuli. The course of these changes was gradual, with some neurons recruited before, others at the time of, and yet others shortly after learning. These observations suggest the hippocampus might contribute to memory by identifying consistencies across experiences that constitute important new associations. PMID- 14550484 TI - Bubbles in the brain? AB - How does the brain learn to control motor actions? 'Bubbles' of activity might exist in the cortex, as implied by a simplified model of the cortical sheet. New results show how these bubbles could enable motor actions to be especially efficient, and that they can be used with little difficulty in decoding them. PMID- 14550485 TI - Mood-dependent memory. AB - Have you ever noticed that when you are in a bad mood the whole world seems to be against you? More negative things seem to happen, and you even remember past episodes of your life in a more negative way than usual. Most of us have experienced this phenomenon, but few will have thought about how this mood might interact with our ability to remember. In a recent paper, Susanne Erk et al. shed light on a possible neural basis for this phenomena. PMID- 14550488 TI - Prometheus to Proust: the case for behavioural criteria for 'mental time travel'. PMID- 14550486 TI - Content, context and cognitive style in mood-memory interactions. Response to Lewis and Critchley. PMID- 14550490 TI - Does colour constancy exist? AB - For a stable visual world, the colours of objects should appear the same under different lights. This property of colour constancy has been assumed to be fundamental to vision, and many experimental attempts have been made to quantify it. I contend here, however, that the usual methods of measurement are either too coarse or concentrate not on colour constancy itself, but on other, complementary aspects of scene perception. Whether colour constancy exists other than in nominal terms remains unclear. PMID- 14550491 TI - 3-D structure perceived from dynamic information: a new theory. AB - Image movement provides one of the most potent two-dimensional cues for depth. From motion cues alone, the brain is capable of deriving a three-dimensional representation of distant objects. For many decades, theoretical and empirical investigations into this ability have interpreted these percepts as faithful copies of the projected 3-D structures. Here we review empirical findings showing that perceived 3-D shape from motion is not veridical and cannot be accounted for by the current models. We present a probabilistic model based on a local analysis of optic flow. Although such a model does not guarantee a correct reconstruction of 3-D shape, it is shown to be consistent with human performance. PMID- 14550492 TI - Imaging a science of mind. AB - I believe that the study of neuroimaging has supported localization of mental operations within the human brain. Most studies have shown a small number of widely distributed brain areas that must be orchestrated to carry out a cognitive task. Although, as in all sciences, there are disagreements, the convergence of results in areas of attention and language in particular seem impressive. Moreover, the anatomical data has helped us to specify the computations that are used by the brain to carry out cognitive tasks. Building upon localization of cognitive operations, imaging methods are being applied to studies of the circuitry, plasticity and individual development of neural networks. Working together with cellular and genetic methods, there is movement towards a more unified view of the role of the human brain in supporting the mind. PMID- 14550493 TI - In two minds: dual-process accounts of reasoning. AB - Researchers in thinking and reasoning have proposed recently that there are two distinct cognitive systems underlying reasoning. System 1 is old in evolutionary terms and shared with other animals: it comprises a set of autonomous subsystems that include both innate input modules and domain-specific knowledge acquired by a domain-general learning mechanism. System 2 is evolutionarily recent and distinctively human: it permits abstract reasoning and hypothetical thinking, but is constrained by working memory capacity and correlated with measures of general intelligence. These theories essentially posit two minds in one brain with a range of experimental psychological evidence showing that the two systems compete for control of our inferences and actions. PMID- 14550494 TI - Multisensory integration, perception and ecological validity. AB - Studies of multimodal integration have relied to a large extent on conflict situations, in which two sensory modalities receive incongruent data concerning one aspect of the source. Exposure to such situations produces immediate crossmodal biases as well as longer lasting aftereffects, revealing recalibrations of data-to-percept matches. In the natural environment, such phenomena might be adaptive, by reducing the perturbing effects of factors like noise or growth-induced changes in receptor organs, and by enriching the percept. However, experimental results generalize to real life only when they reflect automatic perceptual processes, and not response strategies adopted to satisfy the particular demands of laboratory tasks. Here, we focus on this issue and review ways of addressing it that have been developed recently. PMID- 14550496 TI - Summary of major conclusions. PMID- 14550497 TI - Mouse lymphoma thymidine kinase gene mutation assay: International Workshop on Genotoxicity Tests Workgroup report--Plymouth, UK 2002. AB - The Mouse Lymphoma Assay (MLA) Workgroup of the International Workshop on Genotoxicity Tests (IWGT) met on June 28th and 29th, 2002, in Plymouth, England. This meeting of the MLA group was devoted to discussing the criteria for assay acceptance and appropriate approaches to data evaluation. Prior to the meeting, the group conducted an extensive analysis of data from both the microwell and soft agar versions of the assay. For the establishment of criteria for assay acceptance, 10 laboratories (6 using the microwell method and 4 using soft agar) provided data on their background mutant frequencies, plating efficiencies of the negative/vehicle control, cell suspension growth, and positive control mutant frequencies. Using the distribution curves generated from this data, the Workgroup reached consensus on the range of values that should be used to determine whether an individual experiment is acceptable. In order to establish appropriate approaches for data evaluation, the group used a number of statistical methods to evaluate approximately 400 experimental data sets from 10 laboratories entered into a database created for the earlier MLA Workshop held in New Orleans [Environ. Mol. Mutagen. 40 (2002) 292]. While the Workgroup could not, during this meeting, make a final recommendation for the evaluation of data, a general strategy was developed and the Workgroup members agreed to evaluate this new proposed approach using their own laboratory data. This evaluation should lead to a consensus global approach for data evaluation in the near future. PMID- 14550498 TI - In vivo transgenic mutation assays. AB - Transgenic rodent gene-mutation models provide relatively quick and statistically reliable assays for gene mutations in the DNA from any tissue. This report summarizes those issues that have been agreed upon at a previous IWGT meeting [Environ. Mol. Mutagen. 35 (2000) 253], and discusses in depth those issues for which no consensus was reached before. It was previously agreed that for regulatory applications, assays should be based upon neutral genes, be generally available in several laboratories, and be readily transferable. For phage-based assays, five to ten animals per group should be analyzed, assuming a spontaneous mutant frequency (MF) of approximately 3x10(-5) mutants/locus and 125,000-300,000 plaque or colony forming units (pfu or cfu) per tissue per animal. A full set of data should be generated for a vehicle control and two dose groups. Concurrent positive control animals are only necessary during validation, but positive control DNA must be included in each plating. Tissues should be processed and analyzed in a blocked design, where samples from negative control, positive control and each treatment group are processed together. The total number of pfus or cfus and the MF for each tissue and animal are reported. Statistical tests should consider the animal as the experimental unit. Nonparametric statistical tests are recommended. A positive result is a statistically significant dose response and/or statistically significant increase in any dose group compared to concurrent negative controls using an appropriate statistical model. A negative result is a statistically non-significant change, with all mean MFs within two standard deviations of the control. During the current workshop, a general protocol was agreed in which animals are treated daily for 28 consecutive days and tissues sampled 3 days after the final treatment. This recommendation could be modified by reducing or increasing the number of treatments or the length of the treatment period, when scientifically justified. Normally male animals alone are sufficient and normally at least one rapidly proliferating and one slowly proliferating tissue should be sampled. Although, as agreed previously, sequencing data are not normally required, they might provide useful additional information in specific circumstances, mainly to identify and correct for clonal expansion and in some cases to determine a mechanism associated with a positive response. PMID- 14550499 TI - Report from the in vitro micronucleus assay working group. AB - At the Washington "2nd International Workshop on Genotoxicity Testing" (25-26 March 1999) current methodologies and data for the in vitro micronucleus test were reviewed. As a result, guidelines for the conduct of specific aspects of the protocol were developed. Agreement was achieved on the following topics: choice of cells, slide preparation, analysis of micronuclei, toxicity, use of cytochalasin-B, number of doses, and treatment/harvest times [Environ. Mol. Mutagen. 35 (2000) 167]. Because there were a number of important in vitro micronucleus validation studies in progress, it was not possible to design a definitive, internationally harmonized protocol at that time. These studies have now been completed and the data were reviewed at the Plymouth "3rd International Workshop on Genotoxicity Testing" (28-29 June 2002). Data from studies coordinated by the French Society of Genetic Toxicology, Japanese collaborative studies, European pharmaceutical industry validation studies, along with data from Lilly Research Laboratories were used to prepare conclusions on the main aspects of the in vitro micronucleus protocol. In this paper, the consensus agreements on the protocol for performing the in vitro micronucleus assay are presented. The major recommendations concern: 1. Demonstration of cell proliferation: both cell lines and lymphocytes can be used, but demonstration of cell proliferation in both control and treated cells is compulsory for the acceptance of the test. 2. Assessment of toxicity and dose range finding: assessment of toxicity should be performed by determining cell proliferation, e.g. increased cell counts (CC) or population doubling (PD) without cytochalasin B, or e.g. cytokinesis-block proliferation index with cytochalasin-B; and by determining other markers for cytotoxicity (confluency, apoptosis, necrosis) which can provide valuable additional information. 3. Treatment schedules for cell lines and lymphocytes. 4. Choice of positive controls: without S9-mix both a clastogen (e.g. mitomycin C or bleomycin) and an aneugen (e.g. colchicine) should be included as positive controls and a clastogen that requires S9 for activity when S9-mix is used (e.g. dimethylnitrosamine, or cyclophosphamide in those cell types that cannot activate this agent directly). 5. Duplicate cultures and number of cells to be scored. 6. Repeat experiments: in lymphocytes, for each experiment blood from 2 different healthy young and non-smoking donors should be compared. In cell lines, the experiments need only to be repeated if the first one is negative. 7. STATISTICS: statistical significance should not be the sole factor for determining positive results. Biological meaning should serve as a guideline. Examples of statistical analyses are given. PMID- 14550500 TI - Transgenic tumor models for carcinogen identification: the heterozygous Trp53 deficient and RasH2 mouse lines. AB - Genetically altered mouse models (GAMM) for human cancers have been critical to the investigation and characterization of oncogene and tumor suppressor gene expression and function and the associated cancer phenotype. Similarly, several of the mouse models with defined genetic alterations have shown promise for identification of potential human carcinogens and investigation of mechanisms of carcinogen-gene interactions and tumorigenesis. In particular, both the B6.129N5 Trp53 mouse, heterozygous for a p53 null allele, and the CB6F1-RasH2 mouse, hemizygous for the human H-ras transgene, have been extensively investigated. Using 26-week exposure protocols at or approaching the maximum tolerated dose, the summary results to date indicate the potential for GAMM to identify and, possibly, classify chemicals of potential risk to humans using short-term carcinogenicity experiments. This IWGT session focused on: (1) the development of recommendations for genetic/molecular characterization required in animals, tissues, and tumors before and after treatment for identification of presumptive human carcinogens based on the current state of knowledge, (2) identification of data gaps in our current state of knowledge, and (3) development of recommendations for research strategies for further development of our knowledge base of these particular models. By optimization of protocols and identification of significant outcomes and responses to chemical exposure in appropriate short term mechanism-based genetically altered rodent models, strategies for prevention and intervention may be developed and employed to the benefit of public health. PMID- 14550501 TI - Strategy for genotoxicity testing and stratification of genotoxicity test results -report on initial activities of the IWGT Expert Group. PMID- 14550502 TI - Alternate interferon signaling pathways. AB - More than a half a century ago, interferons (IFN) were identified as antiviral cytokines. Since that discovery, IFN have been in the forefront of basic and clinical cytokine research. The pleiotropic nature of these cytokines continues to engage a large number of investigators to define their actions further. IFN paved the way for discovery of Janus tyrosine kinase (JAK)-signal transducing activators of transcription (STAT) pathways. A number of important tumor suppressive pathways are controlled by IFN. Several infectious pathogens counteract IFN-induced signaling pathways. Recent studies indicate that IFN activate several new protein kinases, including the MAP kinase family, and downstream transcription factors. This review not only details the established IFN signaling paradigms but also provides insights into emerging alternate signaling pathways and mechanisms of pathogen-induced signaling interference. PMID- 14550503 TI - A glance at adenosine receptors: novel target for antitumor therapy. AB - Adenosine can be released from a variety of cells throughout the body, as the result of increased metabolic rates, in concentrations that can have a profound impact on the vasculature, immunoescaping, and growth of tumor masses. It is recognized that the concentrations of this nucleoside are increased in cancer tissues. Therefore, it is not surprising that adenosine has been shown to be a crucial factor in determining the cell progression pathway, either during apoptosis or during cytostatic state. From the perspective of cancer, the most important question then may be "Can activation and/or blockade of the pathways downstream of the adenosine receptor contribute to tumor development?" Rigorous examinations of the role of adenosine in in vivo and in vitro systems need to be investigated. The present review therefore proposes multiple adenosine-sustained ways that could prime tumor development together with the critical combinatorial role played by adenosine receptors in taking a choice between proliferation and death. This review proposes that adenosine acts as a potent regulator of normal and tumor cell growth. It is hypothesized that this effect is dependent on extracellular adenosine concentrations, cell surface expression of different adenosine receptor subtypes, and signal transduction mechanisms activated following the binding of specific agonists. We venture to suggest that the clarification of the role of adenosine and its receptors in cancer development may hold great promise for the treatment of chemotherapy in patients affected by malignancies. PMID- 14550504 TI - Interferon-beta in the treatment of relapsing-remitting multiple sclerosis. AB - The role of interferon-beta as a disease-modifying drug (DMD) for the treatment of relapsing-remitting multiple sclerosis (RRMS) is now well established, and its efficacy has been demonstrated unequivocally in large-scale clinical trials. However, current evidence suggests that in order to increase the benefit of therapy, use of an effective drug and dosing regimen should be commenced early in the course of the disease, a finding that places new emphasis on the need for early diagnosis. Indeed, it is now known that MS lesions often develop at a subclinical level and that axonal damage occurs even in the very early stages of the disease. Moreover, such damage may be irreversible, and there is strong evidence to suggest that efficacy lost as a consequence of delay in the onset of treatment or the use of a suboptimal drug regimen cannot be regained. At present, the choice of interferon-beta is complicated by the availability of 3 different products, each with a different dosing regimen. Although the optimal interferon beta dosing regimen for RRMS has been the focus of much discussion, the issues of dose, and particularly dosing frequency, have not been satisfactorily addressed in clinical trials until recently. Over the last 2 years, however, 3 comparative studies of interferon-beta products have been conducted. The results obtained from these recent trials underline the importance of both dose and dosing frequency and indicate that for improved efficacy in RRMS, interferon-beta therapy should be administered frequently at the highest tolerable, and thus most effective, dose. PMID- 14550505 TI - cDNA microarray and proteomic approaches in the study of brain diseases: focus on schizophrenia and Alzheimer's disease. AB - Recent advances in experimental genomics and proteomics, coupled with the wealth of sequence information available for a variety of organisms, have tremendous implications for how biomedical research is performed. Genomic techniques, such as complementary DNA (cDNA) microarrays, currently allow researchers to quickly and accurately quantify vast numbers of potential gene expression changes simultaneously. Modern proteomic techniques allow for the detection and elucidation of protein-protein interactions on a scale and at a speed never before possible. Although hurdles remain, together, these tools open the possibility of enormous change in our ability to analyze and interpret complex biological processes. The field of neuroscience is particularly well suited to analysis with these new techniques, given the complexity of neuronal signaling and the diversity of cellular responses. This review summarizes the major cDNA microarray and proteomic findings of relevance to schizophrenia and Alzheimer's disease (AD) as 2 representative areas of neuroscience research. The potential for these techniques to help unravel the underlying pathology of complex neurological and neuropsychiatric conditions is considerable and warrants continued investigation. PMID- 14550507 TI - A brief history of sudden cardiac death and its therapy. AB - At the end of the 19th century, there was both experimental and clinical evidence that coronary artery obstruction causes ventricular fibrillation and sudden death and that fibrillation could be terminated by electric shocks. The dominant figure at that time was McWilliam, who in 1923 complained that "little attention was given to the new view for many years." This remained so for many decades. It was not until the 1960s that the medical profession became aware of the magnitude of the problem of sudden death and began to install coronary care units where arrhythmias could be monitored and prompt defibrillation could be delivered. This approach was pioneered by Julian in 1961. Milestones that allowed this development were open-chest defibrillation by Beck, closed-chest defibrillation by Zoll, cardiac massage by Kouwenhoven et al., and development of the DC defibrillator by Lown. In 1980, Mirowski et al. implanted the first implantable cardioverter defibrillator (ICD) in a patient. Thereafter, the use of the ICD increased exponentially. Several randomized trials, largely in patients with coronary artery disease and left ventricular dysfunction or in patients with documented lethal arrhythmias, showed beyond doubt that the ICD is superior to antiarrhythmic drug therapy in preventing sudden death, although a number of trials showed no effect. Trials on antiarrhythmic drugs were disappointing. Sodium channel blockers and "pure" potassium channel blockers actually increase mortality, calcium channel blockers have no effect, and, although amiodarone reduces arrhythmic death, it had no effect on total mortality in the 2 largest trials. Only the beta-blockers have been proven to reduce the incidence of sudden death, but their effect appears not to be related to the suppression of arrhythmias but rather to the reduction in sinus rate. Drugs that prevent ischemic events, or lessen their impact, such as anticoagulants, statins, angiotensin-converting enzyme inhibitors, and aldosteron antagonists, all reduce the incidence of sudden death. PMID- 14550506 TI - Why are dendritic cells important in allergic diseases of the respiratory tract? AB - Increasing evidence points to the role of antigen-presenting dendritic cells (DC) in regulating adaptive immune responses. DC are especially sensitive to signals derived from microbes, allergens, and the airway tissue microenvironment, can polarize naive T-cells into either Th1 or Th2 effector cells, and are increasingly recognized as having a central role in the establishment of T-cell memory and tolerance to inhaled antigens. DC form a closely meshed network within the respiratory mucosa and are rapidly recruited from the circulation in response to a variety of proinflammatory stimuli. Studies using animal models have highlighted the role of DC in both initiation and maintenance of allergic airway inflammation. Increased numbers of airway mucosal DC are found in both allergic rhinitis and asthma, and an increasing number of investigators have highlighted important functional differences between DC from atopic and normal individuals. This article reviews recent information on the involvement of DC in the pathogenesis of allergic airway disease and the means by which DC could be exploited as targets for therapy in asthma and allergic rhinitis. PMID- 14550508 TI - Use of an immortalized bovine mammary epithelial cell line (MAC-T) to measure the mitogenic activity of extracts from heifer mammary tissue: effects of nutrition and ovariectomy. AB - The objectives of the experiment were (1) to determine whether MAC-T cells would accurately mimic the previously observed proliferative responses of primary mammary epithelial cells (MEC) to mammary tissue extracts from high and low-fed heifers and (2) to determine whether mammary tissue extracts from ovariectomized (OVX) heifers would have lower mitogenic activity than intact controls. Addition of mammary tissue extracts to cell culture media of MAC-T cells plated on plastic or collagen-coated plastic to a range of concentrations between 1 and 8% resulted in dose-dependent increases in cell proliferation. Furthermore, mammary tissue extracts from low-fed prepubertal heifers aged 9 months, stimulated significantly more proliferation of MAC-T cells, as measured by 3H-thymidine incorporation into DNA than mammary tissue extracts from high-fed heifers (40.6 cpm x 10(3) per well versus 21.9+/-1.8 cpm x 10(3) per well). These observations suggested that MAC-T cells would be a suitable alternative to primary MECs for measuring the mitogenic activity of mammary tissue extracts. Conversely, no difference was observed in the mitogenic activity of mammary tissue extracts from OVX or control heifers. Possibly, MAC-T cells provide a good model for nutrition- but not ovarian-induced changes in mammary growth. Alternatively, that reduction of in vivo mammary development following OVX did not result in reduced mitogenic activity of the mammary tissue extracts emphasizes that heifer mammary development is the result of complex interactions between local growth factors and systemic hormones. PMID- 14550509 TI - Attenuation of PGF2alpha release in ewes infused with the oxytocin antagonist L 368,899. AB - We have investigated the effects of systemic administration of the oxytocin antagonist (OTA) L-368,899 on luteolytic PGF(2alpha) release in ewes. In the first study, carried out in four ovariectomized ewes primed with progesterone to induce responsiveness to oxytocin, 3-h i.v. infusions of 3, 10 and 30 microg/kg/min OTA, carried out on days 12, 14, 16 and 18 in a Latin Square design, resulted in a significant attenuation of the oxytocin induced increase in PGFM concentration at all doses (OTA 139+/-8.3% of pre-oxytocin baseline; control 206.8+/-18.7%; P<0.005). In a further study, continuous infusion of cyclic ewes (n=6) with 10 microg/kg/min OTA from day 13 to day 17 of the cycle resulted in a reduction in both the frequency (OTA 1.0+/-0.4/ewe; control 2.2+/-0.2/ewe; P<0.05) and amplitude (OTA 31.8+/-11.0 pg/ml; control 68.8+/-10.4 pg/ml; P<0.05) of endogenous PGFM episodes compared to control ewes (n=5) measured during daily 8-h sampling windows on days 14-17. This reduction in PGFM concentrations was accompanied by a modest extension in the day of luteolysis (progesterone <0.5 ng/ml) to day 17.5+/-0.4 in the OTA treated group compared with day 16.4+/-0.5 in the control group (P=0.07). The results demonstrate that treatment with OTA caused a significant reduction in episodes of increased PGFM concentration during the period of luteolysis and may provide an approach by which to reduce early pregnancy failure. PMID- 14550510 TI - Plasma levels of cortisol and corticosteroid-binding globulin (CBG) and hepatic CBG mRNA expression in pre- and postnatal pigs. AB - The relationships among hepatic corticosteroid-binding globulin (CBG) mRNA expression and plasma concentrations of cortisol and CBG was evaluated in fetal pigs (n=7-14 per age) on days 50, 70, 80, 90, and 104 of gestation and postnatal pigs (n=8 per age) on days 1, 3, 10, 20, 30, and 40 following birth. In fetal pigs, hepatic CBG mRNA expression was highest (P<0.01) on day 50 as compared to days 90 and 104, exhibiting an overall negative relationship (r=-0.63; P<0.01) with estimated gestation age. Plasma porcine CBG (pCBG) concentration was correlated (r=0.34; P<0.05) with hepatic CBG mRNA level. Plasma cortisol concentrations were not different over this same period. In postnatal pigs, hepatic CBG mRNA expression increased (P<0.01) from days 3 to 40. The pCBG concentration increased (P<0.01) from days 1 (6.1+/-3.4 microg/ml) to 10 (15.1+/ 3.7 microg/ml), while plasma cortisol concentration remained constant. An understanding of the relation between hepatic CBG mRNA and circulating pCBG concentrations may provide insight into the mechanisms determining the bioavailability of cortisol necessary in prenatal development and the conservation of cortisol during postnatal development in the pig. PMID- 14550511 TI - Milking characteristics and their relation to adrenergic receptor mRNA expression and ligand binding in the mammary gland of dairy cows. AB - Stimulation of alpha- and beta-adrenergic receptors in the bovine mammary gland affects milking characteristics such as milk yield and peak flow rate. The aim of this study was to detect possible correlations between milkability, adrenergic receptor binding capacity and receptor expression at the mRNA level. In addition, dose-response relationships of alpha- and beta-adrenergic receptor stimulation were evaluated after application of an alpha- and beta-adrenergic receptor agonist, respectively in different dosages. Density and distribution of adrenergic receptor binding sites in the region around the large mammary ducts were investigated as well as adrenergic receptor mRNA expression. Milk flow of one-quarter was recorded in 10 cows without or with additional alpha- and beta adrenergic receptor stimulation in three dosages each. After slaughter, mammary tissue was taken from the region around the large mammary ducts in the previously investigated quarters. Protein and RNA were extracted for measuring alpha(1)-, alpha(2)-, and beta(2)-adrenergic receptor binding sites and mRNA expression levels by real-time reverse-transcription polymerase chain reaction (RT-PCR). Peak flow rate without additional adrenergic receptor stimulation was negatively correlated with alpha(2)-adrenergic receptor binding (maximal binding capacity, B(max)) and positively correlated with alpha(2)-adrenergic receptor expression at the mRNA level (crossing point (CP) of the real-time PCR). During alpha adrenergic receptor stimulation, there was a positive correlation between milkability and alpha(2)-adrenergic receptor mRNA expression, whereas during beta adrenergic receptor stimulation no correlations were detected. Dose-response relationships were existing during alpha-adrenergic receptor stimulations, but not during beta-adrenergic receptor stimulations at four dosages each including control milking. Significant changes in milk yield and peak flow rate mainly occurred after application of an alpha-adrenergic receptor agonist. In conclusion, high mRNA expression levels or binding capacities of adrenergic receptors do not necessarily lead to according reactions in vivo, concerning milk yield and peak flow rate. To influence milking characteristics, individual reactions of the cow on adrenergic stimulation have to be considered. PMID- 14550512 TI - mRNA expression of components of the IGF system and of GH and insulin receptors in ovaries of broiler breeder hens fed ad libitum or restricted from 4 to 16 weeks of age. AB - An evaluation of the insulin-like growth factors (IGF) system in the ovaries of heavy breeder hens during the growing phase was performed to measure the effect of feed restriction. The transcripts of IGF-I, IGF-II, IGF receptor (IGF-R), two IGF binding proteins (IGFBP-2 and IGFBP-5), GH receptor (GH-R) and insulin receptor (I-R) were measured by RT-PCR at 4, 8, 12 and 16 weeks of age in the ovaries of ad libitum fed and feed restricted hens. The transcripts of all genes screened were detected in all feed regime groups and at all ages sampled. However, no significant overall effects of feed restriction on these transcripts were found despite the marked difference in body growth (200% at 16 weeks of age). Similarly there were no significant age effects except for the expression of GH-R-that showed a decrease after 8 weeks of age (P<0.01). Although feed restriction seems to have a relatively limited effect on the expression of these components of the ovarian IGF system in juveniles, a more detailed determination is necessary to determine possible changes nearer to sexual maturity (16 weeks of age onwards). It is concluded that components of the IGF system, GH or insulin may not be involved in the mechanisms that, through restricted feeding, alter ovarian development and function during growth to enable better reproductive performance during lay. PMID- 14550513 TI - Central effects of rat versus mouse leptin: ingestive behavior and adipose apoptosis. AB - Leptin decreases food intake, body weight and fat mass while sparing lean mass. Although mouse leptin (ML) and rat leptin (RL) are 95.9% identical, our impression from previous studies was that they were not equally potent in rats. Thus, in the present study, 0 microg (vehicle) or 10 microg of ML or RL was injected into the lateral ventricle of rats (eight per treatment) once a day for four consecutive days. Body temperature, body weight, food intake and water intake were measured daily. Intrascapular and perirenal brown fat pads, white fat tissues (retroperitoneal, epididymal and inguinal fat pads) and the gastrocnemius muscle were collected and weighed 24h after the last treatments. Body temperature was not affected by either ML or RL. Both ML and RL caused significant reductions in food intake (P<0.05), and there was no difference between them. ML and RL also similarly inhibited water intake on days 2 and 3 (P<0.05). By day 5 both ML- and RL-treated rats had lost weight (11.6 and 15.4 g, respectively), while vehicle treated rats gained weight (6.8 g). Weights of retroperitoneal and epididymal fat pads, but not other tissues, were reduced similarly by both leptin treatments (P<0.05). However, an increased apoptotic response was detected in the retroperitoneal fat tissue of RL- but not ML-treated rats (P<0.05). These results suggest that RL is more effective than ML in inducing apoptosis in retroperitoneal fat tissue after i.c.v. injection in rats. PMID- 14550515 TI - [Internal medicine: Which missions? And which future?]. PMID- 14550514 TI - Partial cloning and localization of leptin and leptin receptor in the mammary gland of the Egyptian water buffalo. AB - Originally an overall metabolic control was attributed to the leptin hormone, which is produced mainly by the adipose tissue. Recently, leptin gene expression was demonstrated in several additional peripheral tissues. Furthermore, several isoforms of leptin receptor were found both in the central nervous system and in the peripheral tissues. Using reverse transcription and polymerase chain reaction analysis we demonstrate that leptin is expressed both in the adipose tissue and in the lactating mammary gland tissue of Egyptian water buffalo. Our results show that, short and long isoforms of leptin receptor are expressed in buffalo mammary gland tissue. We have partially cloned the buffalo leptin and its short and long isoforms of receptor, which show a high sequence homology to previously published sequences of other mammalian species especially to that of other ruminants. Localization of leptin and its receptor mRNA transcripts, as determined by in situ hybridization procedure, revealed that leptin and its receptor transcripts are expressed specifically in the alveolar epithelial cells of the mammary gland. These morphological data support that leptin could also act as an autocrine and paracrine mediator for mammary gland metabolism and as a facilitator of alveolar epithelial cell activity during lactation. PMID- 14550516 TI - [Management of chronic urticaria: consensus conference's recommendations]. PMID- 14550517 TI - [Common variable immunodeficiency: 17 observations in the adult]. AB - PURPOSE: Common variable immunodeficiency (CVID) is an immune defect characterized by primary hypogammaglobulinemia. Most of the time, clinical manifestations that reveal CVID are recurrent bacterial infections, but auto immune or granulomatous events may occur. METHODS: This retrospective study was conducted on 17 patients fulfilling the classical CVID definition. Lymphocyte activation level was evaluated in 12 patients through HLA-DR expression on lymphocytes subsets. RESULTS: This study includes 17 patients, 7 men and 10 women. The mean age at the first clinical manifestation is 23 years and the mean age at diagnosis is 39 years. Recurrent upper and lower bacterial respiratory tract infections are common to all patients. Abdominal infection due to Mycobacterium avium-intracellulare complex is found in one patient. Digestive events are dominated by chronic diarrhea caused by giardiasis, nodular lymphoid hyperplasia or villous atrophy. Seven patients developed auto-immune conditions (insulin dependent diabetes, idiopathic thrombocytopenic purpura (ITP), rheumatoid arthritis) and 7 patients have a splenomegaly. Non caseating granulomas in the spleen or in lymph node biopsies are found in 3 patients. Ten patients have a T lymphopenia, 2 have a B lymphopenia, 5 have a CD4/CD8 ratio <1, and 6 have T CD4(+) lymphocytes <400/mm(3). The study of HLA-DR expression on lymphocytes subsets shows that 7/12 patients have activated T CD4(+) and/or CD8(+) cells and these patients have auto-immune or tumoral manifestations. The other 5 patients do not have activated T lymphocytes but present with infectious events only. CONCLUSIONS: Our study allows the separation of patients with CVID according to their T lymphocytes activation level. A patient's classification is necessary to define homogeneous groups of patients to perform genetic and functional studies which will probably reveal heterogeneous molecular abnormalities. PMID- 14550518 TI - [Experience of an adherence support for HIV-1 seropositive patients with failure under treatment]. AB - CONTEXT: To be successful highly active antiretroviral therapies need very high levels of adherence to be successful. In need to help patients in dealing with adverse effects, complexity and multiple constraints of these drugs, adherence support programs have been developed. Therefore, the efficacy on viral load of these interventions has not been established in the HIV-1 positive population. The aim of our study was to assess the virological efficacy of this type of program in our institution. METHODS: Non randomized intervention study with a control group. The intervention group was constituted of patients presenting with virological failure who where admitted in the adherence program. The control group (2:1) has been selected in our data base. Factors known to influence viral load have been analyzed by multiple logistic regression. RESULTS: The probability of obtaining a significant decrease of viral load was better at 6 months for patients using the adherence program (OR: 4,48 [2,72-11,8]). This benefit was not sustained at 12 months. The program efficacy varied with the kind of treatment (with or without protease inhibitors), the therapeutic history and the initial viral load. The absence of protease inhibitor, a high initial viral load, small number of previous drugs were linked to viral load decrease. CONCLUSION: The good virological results observed at 6 months in the intervention group are incentive. Long term virological gain and the profile of patients who could make the best of an adherence support program have to be determined. PMID- 14550519 TI - [Neonatal lupus syndrome: review of the literature]. AB - PURPOSE: Neonatal lupus syndrome include skin lesions, hematological and hepatic disorders, and congenital heart block (CHB) in the absence of severe cardiac malformation. This rare disorder is closely linked to transplacental transport of anti-SSA/Ro and anti-SSB/La maternal antibodies. CURRENT KNOWLEDGE AND KEY POINTS: The prevalence of CHB in newborns of anti-Ro/SSA positive women with known connective tissue disease is 2% and the risk of recurrence ranges from 10 to 17%. Skin and systemic lesions are transient, whereas CHB is definitive and is associated with significant morbidity and mortality (estimated at 16-19%). A pacemaker must be implanted in 2/3 of cases. Myocarditis may be associated or may appeared secondarily. Mothers of children with CHB are usually asymptomatic or have Gougerot-Sjogren, or undifferentiated connective tissue disease. Mothers of children with cutaneous manifestations may present with more severe disease and systemic lupus erythematosus. In anti-Ro/SSA positive pregnant women, echocardiograms should be performed at least every 2 weeks from 16 to 24 weeks gestation. Electrocardiogram should be performed for all children. FUTURE PROSPECTS AND PROJECTS: The efficiency of prophylactic treatment of CHB is not established. Therapy for CHB detected in utero is not standardized and involves fluorinated steroids (especially betamethasone). PMID- 14550520 TI - [Severe acute respiratory syndrome: update]. AB - INTRODUCTION: The severe acute respiratory syndrome (SARS) has recently been recognised as a new clinical entity and a new emerging infectious disease that is highly contagious with significant mortality. CURRENT KNOWLEDGE AND KEY POINTS: By the end of 17 June 2003, a total of 8500 cases included 800 deaths had been reported from 30 countries. The cause of SARS has been identified as a new corona virus. In this manuscript, we describe the clinical features, biological findings, virological tests, radiological assessment, outcome and treatments of this infection. FUTURE: Research on specific therapies and vaccines is on-going. PMID- 14550521 TI - [Medical activity of the French internal medicine departments in 2000-2001 from the DRG system]. AB - PURPOSE: Medical activity of the French Internal Medicine Departments has been described until now only by limited declarative surveys. The aim of this study was to describe this activity using the French DRG system. METHODS: A postal survey was conducted using the list of the French Society of Internal Medicine. The same questionnaire was posted to 212 departments (university, public and private hospitals) to collect the 2000 or 2001 data from the DRG system. RESULTS: 192,197 reports were analysed. One hundred and eighty four groups were listed on a theoretical number of 313 medical groups. The first 10 groups concerned common general diseases (pneumonia, anaemia, heart failure, diabetes, stroke.). The main difference between university and non-university hospitals concerned the management of systemic diseases, which was more frequently quoted in university hospitals. Benign and malignant haematological diseases and oncology were frequently quoted in both types of hospitals. CONCLUSION: This study allows for the first time in France to have an objective survey of the medical activity of Internal Medicine Departments. It confirms that these departments have a central role in the hospital management of current diseases but also of haematological diseases and cancer. University departments are more specifically implicated in the management of systemic diseases. PMID- 14550522 TI - [Hemophagocytosis associated with an Escherichia coli sepsis: a case report]. AB - INTRODUCTION: Hemophagocytic lymphohistiocytosis syndrome (HLS) is defined by activated macrophage proliferation. These cells phagocyte the blood elements. This syndrome can be primary as an autosomal recessive disease or secondary to neoplasia, immune diseases or infections-viral, parasitary or bacterian. CASE: Our case concerns an association of HLS and Escherichia coli (E. coli) sepsis in a metastatic prostatic cancer. The evolution was rapidly improved by antibiotics alone. The clinical and biological aspects as well as the differential diagnosis are discussed. CONCLUSION: The HLS is fatal. It can be caused by a severe infection, even an E. coli sepsis. The treatment focused on etiology can be sufficient. PMID- 14550523 TI - [Treatment of a septic patient with acquired haemophilia]. AB - INTRODUCTION: Acquired haemophilia is a rare bleeding diathesis caused by auto immune depletion of factor VIII. It is characterised by spontaneous haemorrhagic syndrome, which can be fatal sometimes. EXEGESIS: A 71 year-old man presents in a dysimmunitary context (rheumatoid arthritis complicates by an acquired haemophilia) a septicemia with a methicillin resistant staphylococcus aureus. At the time of the hospitalization, the patient is febrile (39 degrees C). The activated partial thromboplastin time is very much increased, the level of factor VIII is lowered by 7% and the title of the inhibitor to factor VIII amounts to 140 Bethesda unities. An haematoma of the right root thigh is also noted. In that case, the concomitant presence of septicemia makes difficult the use of immunosuppressive therapy usually recommended to decrease auto-antibody's level. For the management of the septicemia, an adapted antibiotherapy (vancomycin then teicoplanin) is organized to J1. To control haemorrhagic risk, immunoglobulins are prescribed from d12 to d16, without immediate results. Then prednisone is introduced. We observe a very fast decrease of the anticoagulant circulating title with a neat improvement of the clinical state, allowing so to realize a draining puncture of the psoas. This invasive investigation required the use of prothrombinic complex concentrates ((Feiba) in the dose of 80 UI/kg two to three times a day). Biopsy does not show infection source. CONCLUSION: The infection delayed the prescription of immunosuppressive therapy and the surgery. Use of corticoids, following 5 days of intravenous polyvalent immunoglobulin, was the good choice. After 7 weeks of hospitalization the patient has recovered a normal haemostasis results, and a good general state. PMID- 14550524 TI - [Chronic cutaneous ulceration due to Leishmania in an Afghani patient]. PMID- 14550525 TI - [Allergic reaction to methylprednisolone]. PMID- 14550526 TI - [Digestive sarcoidosis: an unusual case revealed by acute ileocecal invagination]. PMID- 14550527 TI - [Acute obstruction of oesophagus by mucilage]. PMID- 14550528 TI - Mouse Tudor Repeat-1 (MTR-1) is a novel component of chromatoid bodies/nuages in male germ cells and forms a complex with snRNPs. AB - Characteristic ribonucleoprotein-rich granules, called nuages, are present in the cytoplasm of germ-line cells in many species. In mice, nuages are prominent in postnatal meiotic spermatocytes and postmeiotic round spermatids, and are often called chromatoid bodies at the stages. We have isolated Mouse tudor repeat-1 (Mtr-1) which encodes a MYND domain and four copies of the tudor domain. Multiple tudor domains are a characteristic of the TUDOR protein, a component of Drosophila nuages. Mtr-1 is expressed in germ-line cells and is most abundant in fetal prospermatogonia and postnatal primary spermatocytes. The MTR-1 protein is present in the cytoplasm of prospermatogonia, spermatocytes, and round spermatids, and predominantly localizes to chromatoid bodies. We show that (1) an assembled form of small nuclear ribonucleoproteins (snRNPs), which usually function as spliceosomal complexes in the nucleus, accumulate in chromatoid bodies, and form a complex with MTR-1, (2) when expressed in cultured cells, MTR 1 forms discernible granules that co-localize with snRNPs in the cell plasm during cell division, and (3) the deletion of multiple tudor domains in MTR-1 abolishes the formation of such granules. These results suggest that MTR-1, which would provide novel insights into evolutionary comparison of nuages, functions in assembling snRNPs into cytoplasmic granules in germ cells. PMID- 14550529 TI - Modifiers of muscle and heart cell fate specification identified by gain-of function screen in Drosophila. AB - The homeobox genes ladybird in Drosophila and their vertebrate counterparts Lbx1 genes display restricted expression patterns in a subset of muscle precursors and are both implicated in diversification of muscle cell fates. In order to gain new insights into mechanisms controlling conserved aspects of cell fate specification, we have performed a gain-of-function (GOF) screen for modifiers of the mesodermal expression of ladybird genes using a collection of EP element carrying Drosophila lines. Amongst the identified genes, several have been previously implicated in cell fate specification processes, thus validating the strategy of our screen. Observed GOF phenotypes have led us to identification of an important number of candidate genes, whose myogenic and/or cardiogenic functions remain to be investigated. Amongst them, the EP insertions close to rhomboid, yan and rac2 suggest new roles for these genes in diversification of muscle and/or heart cell lineages. The analysis of loss and GOF of rhomboid and yan reveals their new roles in specification of ladybird-expressing precursors of adult muscles (LaPs) and ladybird/tinman-positive pericardial cells. Observed phenotypes strongly suggest that rhomboid and yan act at the level of progenitor and founder cells and contribute to the diversification of mesodermal fates. Our analysis of rac2 phenotypes clearly demonstrates that the altered mesodermal level of Rho-GTPase Rac2 can influence specification of a number of cardiac and muscular cell types including those expressing ladybird. Finding that in rac2 mutants ladybird and even skipped-positive muscle founders are overproduced, indicate a new early function for this gene during segregation of muscle progenitors and/or specification of founder cells. Intriguingly, rhomboid, yan and rac2 act as conserved components of Receptor Tyrosine Kinases (RTKs) signalling pathways, suggesting that RTK signalling constitutes a part of a conserved regulatory network governing diversification of muscle and heart cell types. PMID- 14550530 TI - The N-terminal and transmembrane domains of Commissureless are necessary for its function and trafficking within neurons. AB - Commissureless (Comm) is a novel transmembrane molecule necessary both for commissural axons to cross the midline of the Drosophila central nervous system and normal synaptogenesis. Comm is able to reduce cell surface levels of Roundabout (Robo), a receptor for the midline repellent Slit, on commissural axons and unknown inhibitors of synaptogenesis expressed on muscle cells. Comm is expressed dynamically and is found at the cell surface and within intracellular vesicles. Comm can bind Robo and when the proteins are co-expressed Robo is found co-localised with Comm intracellularly. Here we show that the ability of Comm to localise intracellularly and hence regulate Robo surface levels requires sequences in both the N-terminal and transmembrane domains. We also show that Comm can dimerise via its N-terminal domain. Furthermore, absence of the Comm N terminal and transmembrane regions results in the protein being restricted to the neuron soma. PMID- 14550531 TI - mef2c is activated directly by myogenic basic helix-loop-helix proteins during skeletal muscle development in vivo. AB - Skeletal muscle development requires the coordinated expression of numerous transcription factors to control the specification of mesodermal progenitor cells to a muscle fate and the differentiation of those committed myoblasts into functional, contractile muscle. Two families of transcription factors play key roles in these processes. The myogenic basic helix-loop-helix (bHLH) proteins, MyoD and Myf5, are required for myoblast specification, while two members of the same family, myogenin and MRF4, play key roles in myoblast differentiation in vivo. All four members of the myogenic bHLH family are sufficient to dominantly induce myogenesis when introduced into a variety of non-muscle cells in culture, however this function requires the activity of a second family of transcriptional regulators, the myocyte enhancer factor 2 (MEF2) family. MEF2 factors are essential for muscle differentiation, and previous studies have shown that MyoD and MEF2 family members function combinatorially to activate transcription and myogenesis. Consistent with these observations, the majority of skeletal muscle genes require both MyoD and MEF2 family members to activate their transcription. A possible exception to this combinatorial model for activation is suggested by the observation that myogenic bHLH factors may be able to independently activate the expression of MEF2. This raises the question as to how mef2 gene transcription is induced by MyoD factors without cooperative activation by MEF2. During skeletal muscle development, mef2c is the first member of the MEF2 family to be expressed. In this study, we have investigated the regulation of a skeletal muscle-specific enhancer from the mouse mef2c gene using a transgenic approach. We show that mef2c is a direct transcriptional target of the MyoD family in vivo via an essential E box in the skeletal muscle enhancer of mef2c, and we show that mef2c is not a direct target for autoregulation by MEF2. PMID- 14550532 TI - Identification of neurabin II as a novel doublecortin interacting protein. AB - The neuronal migration protein doublecortin (DCX) that associates with microtubules through a tandem DCX repeat, is required for the development of the complex architecture of the human cerebral cortex. Using a yeast two-hybrid screen with Dcx as bait, we have isolated neurabin II/spinophilin, an F-actin binding protein known to play a role in dendritic spine formation. The coiled coil domain of neurabin II binds to a DCX region encompassing the C-terminal portion of the second DCX repeat and the N-terminal portion of the Ser/Pro-rich domain. Immunoprecipitation experiments with brain extracts show that neurabin II and Dcx interact in vivo. Several Dcx constructs that mimic human DCX mutant alleles failed to interact with neurabin II. Since Dcx and neurabin II colocalized in the developing and adult brain, a neurabin II-DCX heterodimer may be involved in neuronal migration and dendritic spine formation. PMID- 14550533 TI - Xenopus nucleosome assembly protein becomes tissue-restricted during development and can alter the expression of specific genes. AB - Nucleosome assembly proteins have been identified in all eukaryotic species investigated to date and their suggested roles include histone shuttle, histone acceptor during transcriptional chromatin remodelling and cell cycle regulator. To examine the role of these proteins during early development we have isolated the cDNA encoding Xenopus NAP1L, raised an antibody against recombinant xNAP1L and examined the expression pattern of this mRNA and protein. Expression in adults is predominantly in ovaries. This maternal protein remains a major component of xNAP1L within the embryo until swimming tadpole stages. xNAP1L mRNA is initially throughout the embryo but by gastrula stages it is predominantly in the presumptive ectoderm. Later, mRNA is detected in the neural crest, neural tube, eyes, tailbud and ventral blood islands. In order to test whether xNAP1L has a potential role in gene regulation we overexpressed this protein in animal pole explants and tested the effect on expression of a series of potential target genes. The mRNA encoding the transcription factor GATA-2 was markedly up regulated by this overexpression. These data support a role for xNAP1L in tissue restricted gene regulation. PMID- 14550534 TI - Congenital diaphragmatic hernia, kidney agenesis and cardiac defects associated with Slit3-deficiency in mice. AB - Slit3 along with Slit1 and Slit2 comprise the Slit family of proteins. The latter two proteins are known to be involved in axon guidance and cell migration during animal development. However, little is know about the functions of Slit3. We created a Slit3-deficient mouse model from an OmniBank ES cell line with a Slit3 allele trapped by insertional mutagenesis to analyze the in vivo functions of this protein. In this model, congenital diaphragmatic hernia is the most obvious phenotype. Herniation was found to be caused by a defective central tendon (CT) of the diaphragm that remained fused with the liver. Electron microscopic analyses of the defective CT revealed disorganized collagen fibrils that failed to form tight collagen bundles. The hearts of Slit3-deficient mice have an enlarged right ventricle. In addition, 20% of homozygous mice also showed a range of kidney defects that include unilateral or bilateral agenesis of the kidney and ureter, or varying degrees of renal hypoplasia. Thus, we concluded that Slit3 is involved in the development of multiple organ systems that include the diaphragm and the kidney. Slit3-deficient mice represent a genetic animal model for physiological and pathological studies of congenital diaphragmatic hernia. PMID- 14550535 TI - Drosophila Perilipin/ADRP homologue Lsd2 regulates lipid metabolism. AB - Many cells store neutral lipids, as triacylglycerol and sterol esters, in droplets. PAT-domain proteins form a conserved family of proteins that are localized at the surface of neutral lipid droplets. Two mammalian members of this family, Perilipin and adipose differentiation-related protein, are involved in lipid storage and regulate lipolysis. Here, we describe the Drosophila PAT-family member Lsd2. We showed that Lsd2 is predominantly expressed in tissues engaged in high levels of lipid metabolism, the fat body and the germ line of females. Ultrastructural analysis in the germ line showed that Lsd2 localizes to the surface of lipid droplets. We have generated an Lsd2 mutant and described its phenotype. Mutant adults have a reduced level of neutral lipid content compared to wild type, showing that Lsd2 is required for normal lipid storage. In addition, ovaries from Lsd2 mutant females exhibit an abnormal pattern of accumulation of neutral lipids from mid-oogenesis, which results in reduced deposition of lipids in the egg. Consistent with its expression in the female germ line, we showed that Lsd2 is a maternal effect gene that is required for normal embryogenesis. This work demonstrates that Lsd2 has an evolutionarily conserved function in lipid metabolism and establishes Drosophila melanogaster as a new in vivo model for studies on the PAT-family of proteins. PMID- 14550536 TI - The role of Suppressor of Hairless in Notch mediated signalling during zebrafish somitogenesis. AB - Suppressor of Hairless (Su(H)) codes for a protein that interacts with the intracellular domain of Notch to activate the target genes of the Delta-Notch signalling pathway. We have cloned the zebrafish homologue of Su(H) and have analysed its function by morpholino mediated knockdown. While there are at least four notch and four delta homologues in zebrafish, there appears to be only one complete Su(H) homologue. We have analysed the function of Su(H) in the somitogenesis process and its influence on the expression of notch pathway genes, in particular her1, her7, deltaC and deltaD. The cyclic expression of her1, her7 and deltaC in the presomitic mesoderm is disrupted by the Su(H) knockdown mimicking the expression of these genes in the notch1a mutant deadly seven. deltaD expression is similarly affected by Su(H) knockdown like deltaC but shows in addition an ectopic expression in the developing neural tube. The inactivation of Su(H) in a fss/tbx24 mutant background leads furthermore to a clear breakdown of cyclic her1 and her7 expression, indicating that the Delta-Notch pathway is required for the creation of oscillation and not only for the synchronisation between neighbouring cells. The strongest phenotypes in the Su(H) knockdown embryos show a loss of all somites posterior to the first five to seven ones. This phenotype is stronger than the known amorphic phenotypes for notch1 (des) or deltaD (aei) in zebrafish, but mimicks the knockout phenotype of RBP-Jkappa gene in the mouse, which is the homologue of Su(H). This suggests that there is some functional redundancy among the Notch and Delta genes. This fact that the first five to seven somites are only weakly affected by Su(H) knockdown indicates that additional genetic pathways may be active in the specification of the most anterior somites. PMID- 14550538 TI - The nuclear protein HMGB1, a new kind of chemokine? AB - The chromosomal protein HMGB1 is now regarded as a proinflammatory cytokine. Importantly, HMGB1 has chemotactic activity suggesting its involvement in the early and late events of the inflammatory reaction. Therefore, HMGB1 has all the hallmarks of a chemokine (chemotactic cytokine). We propose to classify HMGB1 into a new group of proteins unrelated structurally to chemokines but having chemokine-like functions, and to name this class CLF (chemokine-like functions). The CLF class should include other unrelated molecules such as urokinase and its receptor, cytokines macrophage migration inhibitory factor (MIF) and interleukin (IL)-6, anaphylatoxin C5a, ribosomal protein S19, and thioredoxin that have similar chemokine-like activities. This innovative concept may lead to the identification of new therapeutic targets. PMID- 14550537 TI - Diversification and evolution of L-myo-inositol 1-phosphate synthase. AB - L-myo-Inositol 1-phosphate synthase (MIPS, EC 5.5.1.4), the key enzyme in the inositol and phosphoinositide biosynthetic pathway, is present throughout evolutionarily diverse organisms and is considered an ancient protein/gene. Analysis by multiple sequence alignment, phylogenetic tree generation and comparison of newly determined crystal structures provides new insight into the origin and evolutionary relationships among the various MIPS proteins/genes. The evolution of the MIPS protein/gene among the prokaryotes seems more diverse and complex than amongst the eukaryotes. However, conservation of a 'core catalytic structure' among the MIPS proteins implies an essential function of the enzyme in cellular metabolism throughout the biological kingdom. PMID- 14550539 TI - Catalysis, evolution and life. AB - Living organisms are unique in their ability to generate and replicate ordered systems from disordered components. Generation of order, replication of the individual, and evolution of the species all depend on the successful utilization of external energy derived from chemicals and light. The information for reproduction is encoded in nucleic acids, but evolution depends on a limited variability in replication, and proceeds through the selection of individuals with altered biochemistry. Essentially all biochemistry is catalyzed; therefore, altered biochemistry implies altered or new catalysts. In that sense catalysis is the medium of evolution. We propose that a basic property of enzymes, at least as fundamental as reaction rate enhancement, is to adjust the reaction path by altering and eventually optimizing the reversible interchange of chemical, electrical and mechanical energy among themselves and their reactants. PMID- 14550540 TI - 2-Cyclopenten-1-one and prostaglandin J2 reduce restenosis after balloon angioplasty in rats: role of NF-kappaB. AB - The aim of this study was to evaluate, using a rat model of balloon angioplasty, whether prostaglandin (PG) J(2) and 2-cyclopenten-1-one are able to reduce restenosis. We found that both PGJ(2) and 2-cyclopenten-1-one, administered by local application on carotid arteries, caused a dose-dependent inhibition of neointimal formation. Furthermore, both agents prevented vascular negative remodeling. The effect of these compounds on restenosis was correlated with an inhibition of nuclear factor-kappaB (NF-kappaB) activation as well as of intercellular adhesion molecule-1 (ICAM-1) protein expression in injured carotid arteries of control animals. Our results show that cyclopentenone PGs and their derivatives reduce restenosis and may have therapeutic relevance for the prevention of human restenosis. PMID- 14550541 TI - AXR1 is involved in BR-mediated elongation and SAUR-AC1 gene expression in Arabidopsis. AB - Limited information is available concerning the interactions between the brassinosteroid (BR) and auxin signaling pathways. The expression pattern of the SAUR-AC1 gene, an early auxin-inducible gene in Arabidopsis, was studied in response to brassinolide (BL), in the presence of a BR-biosynthesis inhibitor, in a BR-deficient mutant, and in combination with auxin. The results suggested that the SAUR-AC1 gene is regulated by BRs independently of auxin levels, and that it is important in BR-mediated elongation. The axr1 (auxin insensitive 1) mutant was less sensitive to BL-induced elongation and BL-induced SAUR-AC1 expression, suggesting that a ubiquitin ligase-mediated system is involved in BR-mediated elongation. PMID- 14550542 TI - Viral dsRNA activates mucin transcription in airway epithelial cells. AB - Double-stranded (ds) RNA is a biologically active component of many viruses including rhinoviruses infecting the upper respiratory tract. Mucus production is a common symptom of such infections. Here, we show that mucin, the glycoprotein subunit of mucus gels, is transcriptionally upregulated in an NF-kappaB- and p38 dependent manner when homogeneous cultures of epithelial cells are exposed to dsRNA. Furthermore, upstream of p38 in this system, dsRNA stimulates the extracellular release of ATP and activation of cell surface ATP receptors, which are G protein-coupled. This results in the stimulation of phospholipase C and protein kinase C. These findings suggest that ATP receptor antagonists could be used to modulate mucus production induced by virus. PMID- 14550543 TI - Conserved signature proposed for folding in the lipocalin superfamily. AB - We systematically identify a group of evolutionarily conserved residues proposed for folding in a model beta-barrel superfamily, the lipocalins. The nature of conservation at the structural level is defined and we show that the conserved residues are involved in a network of interactions that form the core of the fold. Exploratory kinetic studies are conducted with a model superfamily member, human serum retinol-binding protein, to examine their role. The present results, coupled with key experimental studies conducted with another lipocalin beta lactoglobulin, suggest that the evolutionarily conserved regions fold on a faster folding time-scale than the non-conserved regions. PMID- 14550544 TI - A novel yeast expression system for the overproduction of quality-controlled membrane proteins. AB - Saturation of the cell's protein folding capacity and accumulation of inactive incompletely folded protein often accompanying the overexpression of membrane proteins (MPs) presents an obstacle to their efficient purification in a functional form for structural studies. We present a novel strategy for optimization of functional MP expression in Saccharomyces cerevisiae. This approach exploits the unfolded protein response (UPR) pathway, a stress signaling mechanism that senses the accumulation of unfolded proteins in the endoplasmic reticulum. We demonstrate that a high level of UPR induction upon expression of a MP reflects impaired functional expression of that protein. Tuning the expression level of the protein so as to avoid or minimize UPR induction results in its increased functional expression. UPR status can therefore serve as a proxy variable for the extent of impaired expression of a MP that may even be applicable in the absence of knowledge of the protein's biological function. PMID- 14550545 TI - B cell receptor-mediated nuclear fragmentation proceeds in WEHI 231 cells in the absence of detectable DEVDase and FRase activity. AB - Crosslinking of the WEHI 231 lymphoma B cell receptor (BCR) leads to growth arrest followed by apoptosis. In a study of the role of lysosomal cysteine proteinases in BCR-mediated apoptosis we provide evidence that commitment to apoptosis correlates with a time-dependent increase in caspase and cathepsin activities. We also show that activation of cathepsins is a caspase-independent process, and caspase cascade activation is independent of lysosomal endopeptidases. BCR-induced nuclear fragmentation was not prevented, but rather delayed in the absence of detectable caspase and cathepsin activities, suggesting that BCR-driven apoptosis of these cells may use an alternative proteolytic mechanism independent of caspases and cathepsins. PMID- 14550546 TI - Nitric oxide-dependent cytoskeletal changes and inhibition of endothelial cell migration contribute to the suppression of angiogenesis by RAD50 gene transfer. AB - Previous reports showed that human RAD50 (hRAD50) gene delivery induced regression of an experimental rat tumor and porcine neointimal hyperplasia. In this study, we examined the effects of hRAD50 on the morphological changes and migration of endothelial cells (EC) as possible mechanisms by which hRAD50 might block angiogenesis. Quantitative image analysis revealed significant inhibition of the number and total area of blood vessels in rat tumor tissues following hRAD50 gene delivery. hRAD50 distorted actin and tubulin arrangements, and significantly reduced the F/G-actin ratio and increased the nitric oxide (NO) production in the primary cultured human EC. These effects were blocked by pretreatment with L-NAME (N(G)-nitro-L-arginine-methyl ester), a NO synthase inhibitor. FACScan analysis showed that NO was involved in the necrosis and apoptosis of EC by hRAD50. hRAD50 also inhibited EC migration in an in vitro wound-healing model. These results indicate that NO-dependent cytoskeletal changes and inhibition of EC migration contribute to the suppression of angiogenesis by hRAD50 delivery in vivo. PMID- 14550547 TI - Phosphorylation of insulin receptor substrate-1 serine 307 correlates with JNK activity in atrophic skeletal muscle. AB - c-Jun NH(2)-terminal kinase (JNK) has been shown to negatively regulate insulin signaling through serine phosphorylation of residue 307 within the insulin receptor substrate-1 (IRS-1) in adipose and liver tissue. Using a rat hindlimb suspension model for muscle disuse atrophy, we found that JNK activity was significantly elevated in atrophic soleus muscle and that IRS-1 was phosphorylated on Ser(307) prior to the degradation of the IRS-1 protein. Moreover, we observed a corresponding reduction in Akt activity, providing biochemical evidence for the development of insulin resistance in atrophic skeletal muscle. PMID- 14550548 TI - Effects of glycerol and high temperatures on structure and function of phycobilisomes in Synechocystis sp. PCC 6803. AB - The effects of glycerol and high temperatures on structure and function of phycobilisomes (PBSs) in vivo were investigated in a chlL deletion mutant of the cyanobacterium Synechocystis sp. PCC 6803. When the mutant was grown under light activated heterotrophic growth conditions, it contained intact and functional PBSs, but essentially no chlorophyll and photosystems. So the structural and functional changes of the mutant PBSs in vivo can be handily detected by measurement of low temperature (77 K) fluorescence emission spectra. High concentration glycerol induced an obvious disassembly of PBSs and the dissociation of phycocyanins in the rod substructures into their oligomers and monomers. PBSs also disassembled at high temperatures and allophycocyanins were more sensitive to heat stress than phycocyanins. Our results demonstrate that the chlL(-) mutant strain is an advantageous model for studying the mechanisms of assembly and disassembly of protein complexes in vivo. PMID- 14550549 TI - A study of metabolic compartmentation in the rat heart and cardiac mitochondria using high-resolution magic angle spinning 1H NMR spectroscopy. AB - High-resolution magic angle spinning (MAS) (1)H nuclear magnetic resonance (NMR) spectroscopy is increasingly being used to monitor metabolic abnormalities within cells and intact tissues. Many toxicological insults and metabolic diseases affect subcellular organelles, particularly mitochondria. In this study high resolution (1)H NMR spectroscopy was used to examine metabolic compartmentation between the cytosol and mitochondria in the rat heart to investigate whether biomarkers of mitochondrial dysfunction could be identified and further define the mitochondrial environment. High-resolution MAS spectra of mitochondria revealed NMR signals from lactate, alanine, taurine, choline, phosphocholine, creatine, glycine and lipids. However, spectra from mitochondrial extracts contained additional well-resolved resonances from valine, methionine, glutamine, acetoacetate, succinate, and aspartate, suggesting that a number of metabolites bound within the mitochondrial membranes occur in 'NMR invisible' environments. This effect was further investigated using diffusion-weighted measurements of water and NMR spectroscopy during state 2 and state 3 respiration. State 3 respiration caused a decrease in the resonance intensity of endogenous succinate compared with state 2 respiration, suggesting that coupled respiration may also modulate the NMR detection of metabolites within mitochondria. PMID- 14550550 TI - Photophysical properties of Prochlorococcus marinus SS120 divinyl chlorophylls and phycoerythrin in vitro and in vivo. AB - Prochlorococcus marinus SS120 is an ecologically important and biochemically intriguing marine cyanobacterium. In addition to divinyl chlorophylls (DV-Chls) a and b it possesses a particular form of phycoerythrin (PE), but no other phycobilins and therefore no complete phycobilisomes. Here, a spectroscopic characterisation of these DV-Chls and PE is provided. Comparison of fluorescence quantum yields, excited state lifetimes and absorption characteristics indicate similar light-harvesting properties of the DV-Chls as their monovinyl counterparts. PE, which is present only in tiny amounts, was purified and considerably enriched. A phycourobilin to phycoerythrobilin ratio of 3:1 chromophores per (alphabeta) PE monomer is suggested. The in vitro fluorescence lifetime of PE is 1.74 ns. In vivo time-resolved fluorescence measurements with synchrotron radiation were used to investigate the possible role of PE in light harvesting. The fluorescence decay time for PE is about 550 ps, indicating an unusually slow excitation energy transfer. The decay time slowed to 1 ns after addition of glycerol to cell cultures. The contribution of PE to total light harvesting capacity was estimated to be about one (alphabeta) PE monomer per 330 DV-Chl b molecules. Thus, the capacity of PE to function primarily as a photosynthetic light-harvesting pigment in P. marinus SS120 is low. PMID- 14550551 TI - Development of a fluorescent nanosensor for ribose. AB - To analyze ribose uptake and metabolism in living cells, nanosensors were engineered by flanking the Escherichia coli periplasmic ribose binding protein with two green fluorescent protein variants. Following binding of ribose, fluorescence resonance energy transfer decreased with increasing ribose concentration. Five affinity mutants were generated covering binding constants between 400 nM and 11.7 mM. Analysis of nanosensor response in COS-7 cells showed that free ribose accumulates in the cell and is slowly metabolized. Inhibitor studies suggest that uptake is mediated by a monosaccharide transporter of the GLUT family, however, ribose taken up into the cell was not or only slowly released, indicating irreversibility of uptake. PMID- 14550552 TI - Fluorescence visualization of ultraviolet-sensitive cone photoreceptor development in living zebrafish. AB - Cone photoreceptor cells of fish retinae are arranged in a highly organized fashion. However, the molecular mechanisms underlying photoreceptor development and retinal pattern formation are largely unknown. Here we established transgenic lines of zebrafish carrying green fluorescent protein (GFP) cDNA with the 5.5-kb upstream region of the ultraviolet-sensitive cone opsin gene (SWS1). In the transgenic fish, GFP gene expression proceeded in the same spatiotemporal pattern as SWS1 in the retinae of embryos. In the adult retina, GFP expression was observed throughout the short single cone (SSC) layer where SWS1 is specifically expressed. Therefore, the transgenic fish provides an excellent genetic background to study retinal pattern formation, photoreceptor determination and differentiation, and factors regulating these processes and SSC-specific expression of SWS1. PMID- 14550553 TI - The N-terminus is unstructured, but not dynamically disordered, in the complex between HK022 Nun protein and lambda-phage BoxB RNA hairpin. AB - The Nun protein of lambdoid phage HK022 excludes lambda-phage superinfection by blocking expression of genes downstream from the lambda nut sequences. Heteronuclear NMR studies have been performed on a Nun peptide comprising residues 1-49 bound to the nutR BoxB RNA. The pattern of (13)C chemical shifts indicates that the arginine-rich motif of Nun forms an induced alpha-helix, consisting of residues 23-43, when bound to BoxB RNA, consistent with the structure of a shorter (residues 22-44) Nun peptide/BoxB RNA complex [Faber, C., Scharpf, M., Becker, T., Sticht, H. and Rosch (2001) J. Biol. Chem. 276, 32064 32070]. The N-terminal extension, residues 1-22, does not show chemical shifts or nuclear Overhauser effects characteristic of stable secondary structure. Nonetheless, (15)N relaxation measurements indicate that this region is not completely disordered, as expected for a random coil peptide. Restriction of conformation flexibility in the N-terminal extension of Nun may be important for binding to other target molecules involved in transcription termination. PMID- 14550554 TI - Sequence of Escherichia coli O128 antigen biosynthesis cluster and functional identification of an alpha-1,2-fucosyltransferase. AB - O128 is one of the most common atypical enteropathogenic Escherichia coli isolated from diarrhea patients worldwide. The primary structure of E. coli O128 repeat units has previously been determined as -->3)-beta-D-GalNAc-(1-->4)-alpha D-Gal-(1-->3)-beta-D-GalNAc-(1-->6)-[alpha-L-Fuc-(1-->2)]-beta-D-Gal-(1--> pentasaccharide. Here we report the complete sequencing of E. coli O128 antigen biosynthesis gene cluster and its flanking regions. Comparative sequence analysis revealed the expected O128 antigen process genes, GDP-fucose biosynthesis genes and four potential glycosyltransferase genes responsible for the assembly of E. coli O128 antigen repeats. WbsJ was shown to encode an alpha-1,2 fucosyltransferase by enzymatic assays and nuclear magnetic resonance spectroscopy analysis. PMID- 14550555 TI - Upregulation of the matrix metalloproteinase-1 gene by the Ewing's sarcoma associated EWS-ER81 and EWS-Fli-1 oncoproteins, c-Jun and p300. AB - The mechanisms of action of Ewing's sarcoma (EWS) associated EWS-ETS oncoproteins have largely remained unresolved. Here, we analyzed how two EWS-ETS proteins, EWS ER81 and EWS-Fli-1, in vitro activate the matrix metalloproteinase (MMP)-1 promoter that is upregulated in a subset of EWSs. EWS-ER81 and EWS-Fli-1 interact with and thereby activate the MMP-1 promoter, which is potentiated by the cofactor p300 and the proto-oncoprotein c-Jun. Further, EWS-ER81 binds to c-Jun in vitro and in vivo. The interaction between c-Jun, p300 and EWS-ER81 or EWS-Fli 1 may also be relevant to the regulation of other yet-to-be-identified genes that are responsible for EWS formation. PMID- 14550556 TI - Decrease in the efficiency of the electron donation to tyrosine Z of photosystem II in an SQDG-deficient mutant of Chlamydomonas. AB - Photosystem (PS) II activity of a sulfoquinovosyl diacylglycerol (SQDG)-deficient mutant (hf-2) of Chlamydomonas was partially decreased compared with that of wild type. The susceptibility to 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) was also modified in the mutant. Photometric measurements in the isolated thylakoid membranes of hf-2 revealed that the lowered activity in the mutant was derived from a decrease in the efficiency of the electron donation from water to tyrosine Z, not from the efficiency of the electron transport from Q(A) to Q(B). This result was confirmed by the decay kinetics of chlorophyll fluorescence determined in vivo. We conclude that SQDG contributes to maintaining the conformation of PSII complexes, particularly that of D1 polypeptides, which are necessary for maximum activities in Chlamydomonas. PMID- 14550557 TI - Characterization of Arabidopsis secretory phospholipase A2-gamma cDNA and its enzymatic properties. AB - Plant secretory phospholipases A(2) (sPLA(2)s) probably play important roles in phospholipid signaling based on the data reported from other organisms, but their functions are poorly understood because of the lack of cloned sPLA(2) genes. In this study, we cloned and characterized an Arabidopsis secretory phospholipase A(2)-gamma (AtsPLA(2)-gamma) cDNA, and examined its enzymatic properties. The recombinant protein of AtsPLA(2)-gamma showed maximal enzyme activity at pH 8.0, and required Ca(2+) for activity. Moreover, AtsPLA(2)-gamma showed sn-2 position specificity but no prominent acyl preference, though it showed head group specificity to phosphatidylethanolamine rather than to phosphatidylcholine. AtsPLA(2)-gamma was found to predominate in the mature flower rather than in other tissues, and subcellular localization analysis confirmed that AtsPLA(2) gamma is secreted into the intercellular space. PMID- 14550558 TI - TIGRINA d, required for regulating the biosynthesis of tetrapyrroles in barley, is an ortholog of the FLU gene of Arabidopsis thaliana. AB - Regulation of tetrapyrrole biosynthesis in higher plants has been attributed to negative feedback control of steps prior to delta-aminolevulinic acid (ALA) formation. One of the first mutants with a defect in this control had been identified in barley. The tigrina (tig) d mutant accumulates 10-15-fold higher amounts of protochlorophyllide than wild type, when grown in the dark. The identity of the TIGRINA d protein and its mode of action are not known yet. Initially this protein had been proposed to act as a repressor of genes that encode enzymes involved in early steps of ALA formation, but subsequent attempts to confirm this experimentally failed. Here we demonstrate that the TIGRINA d gene of barley is an ortholog of the FLU gene of Arabidopsis thaliana. The FLU protein is a nuclear-encoded plastid protein that plays a key role in negative feedback control of chlorophyll biosynthesis in higher plants. Sequencing of the FLU gene of barley revealed a frame shift mutation in the FLU gene of the tig d mutant that results in the loss of two tetratricopeptide repeats that in the FLU protein of Arabidopsis are essential for its biological activity. This mutation cosegregates strictly with the tigrina phenotype within the F1 population of a heterozygous tig d mutant, thus providing additional support for the flu gene being responsible for the tigrina phenotype of barley. PMID- 14550559 TI - Poly-L-lysine enhances the protein disaggregation activity of ClpB. AB - The Hsp100 protein ClpB is a member of the AAA+ protein family that mediates the solubilization of aggregated proteins in cooperation with the DnaK chaperone system. Unstructured polypeptides such as casein or poly-L-lysine have been shown to stimulate the ATPase activity of ClpB and thus may both act as substrates. Here we compared the effects of alpha-casein and poly-L-lysine on the ATPase and chaperone activities of ClpB. alpha-Casein stimulated ATP hydrolysis by both AAA domains of ClpB and inhibited the ClpB-dependent solubilization of aggregated proteins if present in excess. In contrast, poly-L-lysine stimulated exclusively the ATPase activity of the second AAA domain and increased the disaggregation activity of ClpB. Thus poly-L-lysine does not act as substrate, but rather represents an effector molecule, which enhances the chaperone activity of ClpB. PMID- 14550560 TI - Extremely rapid turnover of S-adenosylmethionine decarboxylase in Crithidia fasciculata. AB - The activity of S-adenosylmethionine decarboxylase (AdoMetDC) in Crithidia fasciculata was shown to be correlated to the growth of the parasite. An increase in activity was observed during exponential growth. Inhibition of protein synthesis induced an extremely rapid decay of AdoMetDC activity. The half-life of the enzyme was estimated to be about 3 min, which is the shortest half-life ever recorded for an eukaryotic AdoMetDC. The reduction in AdoMetDC activity was correlated with a decrease in AdoMetDC protein content, demonstrating a rapid turnover of the enzyme. No polyamine-mediated feedback regulation of AdoMetDC was observed in the parasite. PMID- 14550562 TI - The mouse sino-atrial node expresses both the type 2 and type 3 Ca(2+) release channels/ryanodine receptors. AB - Ca(2+) released from intracellular Ca(2+) stores is shown to be involved in pacemaker activity in the sino-atrial (SA)-node. However, little is known about the molecular identity of the Ca(2+) release channel/ryanodine receptor (RYR) involved in pacemaker activity. We examined the mRNA distribution of three different RYR isoforms (RYR1, RYR2, and RYR3) in the mouse SA-node. RNase protection assay and in situ hybridization revealed that RYR2 mRNA expresses widely in the heart including the SA-node, while RYR3 mRNA expression is limited to the SA-node and to the right atrium. Thus, not only RYR2 but also RYR3 may participate in pacemaker activity. PMID- 14550561 TI - Endotoxin-neutralizing effects of histidine-rich peptides. AB - Inflammatory responses of human peripheral blood monocytes to the Gram-negative endotoxin lipopolysaccharide (LPS) are enhanced by structurally diverse substances, such as anionic polysaccharides or cationic polypeptides. Only a few substances are known to effectively blunt LPS-induced monocyte activation. We now show that synthetic poly-L-histidine (Hn) binds to LPS and abrogates the release of the proinflammatory cytokine interleukin-8 (IL-8) in LPS-stimulated human whole blood. LPS-induced stimulation of monocytes was strictly pH-dependent with only minor amounts of IL-8 secreted in acidic blood. Maximum levels of IL-8 secretion occurred at a strongly basic pH. Hn inhibition of the release of IL-8 from LPS-stimulated monocytes was observed under acidic, neutral and physiological conditions. With increasing alkalosis, the effectiveness of Hn was gradually lost, suggesting that protonated, but not deprotonated, Hn was effective in inhibiting LPS-induced monocyte responses. Histidine-rich protein 2 from the malaria parasite, Plasmodium falciparum, inhibited the ability of LPS to evoke an inflammatory response in CD14-transfected THP-1 cells. Further, a short synthetic peptide derived from human histidine- and proline-rich glycoprotein also exhibited LPS-inhibitory effects in CD14 transfectants. Taken together, these observations demonstrate the capacity of histidine-rich peptides, irrespective of their origin, to neutralize LPS-induced proinflammatory host responses. PMID- 14550563 TI - Mildly oxidised LDL induces more macrophage death than moderately oxidised LDL: roles of peroxidation, lipoprotein-associated phospholipase A2 and PPARgamma. AB - Death of macrophages and smooth muscle cells (SMC) can lead to progression of atherosclerosis. Mildly oxidised low-density lipoprotein (mildly-oxLDL) induced more overall death and apoptosis than moderately oxidised LDL, in human monocyte macrophages (HMM). Mildly-oxLDL also induced more overall death in human SMC than did moderately-oxLDL. Mildly-oxLDL contained more hydroperoxides, but less oxysterol, malondialdehyde and negative charge than moderately-oxLDL. Specific inhibition of lipoprotein-associated phospholipase A(2) (by SB222657) diminished death induction in HMM by both oxLDL types. Peroxisome proliferator-activated receptor gamma (PPARgamma) antagonist (GW9662) and agonist (ciglitazone) experiments suggested that non-hydrolysed, oxidised phospholipids in oxLDL activate PPARgamma as a cellular defence mechanism. These results may be relevant to LDL oxidation within atherosclerotic plaques and may suggest strategies for combating atherosclerosis progression. PMID- 14550564 TI - The ubiquitin-like domain of Herp is involved in Herp degradation, but not necessary for its enhancement of amyloid beta-protein generation. AB - Herp is an endoplasmic reticulum (ER)-stress-inducible membrane protein, which has a ubiquitin-like domain (ULD). However, its biological function is as yet unknown. Previously, we reported that a high expression level of Herp in cells increases the generation of amyloid beta-protein (Abeta) and that Herp interacts with presenilin (PS). Here, we addressed the role of the ULD of Herp in Abeta generation and intracellular Herp stability. We found that the ULD is not essential for the enhancement of Abeta generation by Herp expression and the interaction of Herp with PS, but is involved in the rapid degradation of Herp, most likely via the ubiquitin/proteasome pathway. Thus, the ULD of Herp most likely plays a role in the regulation of the intracellular level of Herp under ER stress. PMID- 14550565 TI - [3H]BHDP as a novel and selective ligand for sigma1 receptors in liver mitochondria and brain synaptosomes of the rat. AB - The binding profile of [(3)H]BHDP ([(3)H]N-benzyl-N'-(2-hydroxy-3,4 dimethoxybenzyl)-piperazine) was evaluated. [(3)H]BHDP labelled a single class of binding sites with high affinity (K(d)=2-3 nM) in rat liver mitochondria and synaptic membranes. The pharmacological characterization of these sites using sigma reference compounds revealed that these sites are sigma receptors and, more particularly, sigma1 receptors. Indeed, BHDP inhibited [(3)H]pentazocine binding, a marker for sigma1 receptors, with high affinity in a competitive manner. BHDP is selective for sigma1 receptors since it did not show any relevant affinity for most of the other receptors, ion channels or transporters tested. Moreover, in an in vitro model of cellular hypoxia, BHDP prevented the fall in adenosine triphosphate (ATP) levels caused by 24 h hypoxia in cultured astrocytes. Taken together, these results demonstrate that [(3)H]BHDP is a potent and selective ligand for sigma1 receptors showing cytoprotective effects in astrocytes. PMID- 14550566 TI - Control of mitochondrial protein import by pH. AB - Protein import into mitochondria is inhibited by protons (IC(50) pH 6.5). The channels of the import machinery were examined to further investigate this pH dependence. TOM and TIM23 are the protein translocation channels of the mitochondrial outer and inner membranes, respectively, and their single channel behaviors at various pHs were determined using patch-clamp techniques. While not identical, increasing H(+) concentration decreases the open probability of both TIM23 and TOM channels. The pattern of the pH dependences of protein import and channel properties suggests TIM23 open probability can limit import of nuclear encoded proteins into the matrix of yeast mitochondria. PMID- 14550567 TI - Correlation of death modes of photosensitized cells with intracellular ATP concentration. AB - The impact of intensity of glycolysis and oxidative phosphorylation on death of photosensitized murine hepatoma MH22 cells in vitro has been investigated. Cells photosensitized with meso-tetra(4-sulfonatophenyl)-porphine localized to lysosomes died mostly by necrosis, and the mode of cell death did not depend on the energy metabolism. Photosensitization with 5-aminolevulinic acid-stimulated endogenous porphyrins localized mainly in mitochondria or 5,10,15,20-tetrakis(m hydroxyphenyl)-chlorine localized to cell membranes, including mitochondria, led to cell death mostly by apoptosis. In this case, the mode of cell death depended on the medium: under conditions unfavorable to glycolysis the ratio apoptosis/necrosis decreased significantly. PMID- 14550568 TI - Regulation of glycogen synthase kinase 3 in human platelets: a possible role in platelet function? AB - In this study we show that both glycogen synthase kinase 3 (GSK3) isoforms, GSK3alpha and GSK3beta, are present in human platelets and are phosphorylated on Ser(21) and Ser(9), respectively, in platelets stimulated with collagen, convulxin and thrombin. Phosphorylation of GSK3alpha/beta was dependent on phosphoinositide 3-kinase (PI3K) activity and independent of platelet aggregation, and correlated with a decrease in GSK3 activity that was preserved by pre-incubating platelets with PI3K inhibitor LY294002. Three structurally distinct GSK3 inhibitors, lithium, SB415286 and TDZD-8, were found to inhibit platelet aggregation. This implicates GSK3 as a potential regulator of platelet function. PMID- 14550569 TI - pkn22 (alr2502) encoding a putative Ser/Thr kinase in the cyanobacterium Anabaena sp. PCC 7120 is induced by both iron starvation and oxidative stress and regulates the expression of isiA. AB - In cyanobacteria, the isiA gene is required for cell adaptation to oxidative damage caused by the absence of iron. We show here that a putative Ser/Thr kinase gene, pkn22 (alr2052), is activated by iron deficiency and oxidative damage in Anabaena sp. PCC 7120. A pkn22 insertion mutant is unable to grow when iron is limiting. pkn22 regulates the expression of isiA (encoding CP43'), but not of isiB (encoding flavodoxin) and psbC (CP43). Fluorescence measurement at 77 K reveals the absence of the typical signature of CP43' associated with photosystem I in the mutant under iron-limiting conditions. We propose that Pkn22 is required for the function of isiA/CP43' and constitutes a regulatory element necessary for stress response. PMID- 14550570 TI - BRCA1 interacts with FHL2 and enhances FHL2 transactivation function. AB - Germ-line mutations in BRCA1 are associated with an increased lifetime risk of developing breast and/or ovarian tumors. The BRCA1 gene product is a 220-kDa protein that contains a tandem of two BRCA1 C-terminal (BRCT) domains required for transcription. In an attempt to understand how BRCA1 exerts its function through BRCT domains, we search for partners of the BRCT domains of BRCA1. Using the yeast two-hybrid system, we identified the four and a half LIM-only protein 2 (FHL2) as a novel BRCA1 interacting protein. We demonstrate that BRCA1 and FHL2 can physically associate in vitro, in yeast, and in human cells. BRCA1 interacted with FHL2 through its second BRCT domain and the interaction of FHL2 with BRCA1 requires the last three LIM domains of FHL2. BRCA1 enhanced FHL2-mediated transcriptional activity in transient transfections. Tumor-derived transactivation-deficient BRCA1 mutants showed a reduced ability to enhance transactivation by FHL2. Lack of BRCA1 binding sites in the FHL2 completely abolished the FHL2 transactivation function. Reverse transcription polymerase chain reaction analysis showed that FHL2 mRNA levels may be downregulated in many breast cancer cell lines. These results suggest that the BRCA1-FHL2 interaction may be involved in transcriptional regulation and play a significant role in cancer cell growth. PMID- 14550571 TI - Ubiquitin activated tumor necrosis factor receptor associated factor-6 (TRAF6) is recycled via deubiquitination. AB - Ubiquitination of intermediates in the interleukin-1 (IL-1) signaling cascade plays an important role in activation and regulation of the pathway. Both IL-1 receptor associated kinase-1 (IRAK1) and inhibitor of nuclear factor kappaB-alpha (IkappaBalpha) are rapidly ubiquitinated and degraded. Tumor necrosis factor associated factor-6 (TRAF6) is an ubiquitin ligase that is activated by ubiquitination and a signaling intermediate between IRAK1 and IkappaBalpha. It is unknown whether activated TRAF6 is subsequently degraded. We show that in liver cells IL-1 stimulates TRAF6 poly-ubiquitination. In less than 1 h levels of non modified TRAF6 return to levels near those observed prior to activation. TRAF6 cannot be reactivated in cells which have been pretreated with IL-1. This observation correlates with decreased levels of IRAK1 in IL-1 pretreated cells. The re-establishment of non-modified TRAF6 levels following activation does not require de novo protein synthesis, strongly suggesting that TRAF6 is recycled via deubiquitination. This indicates a unique mechanism of regulation of TRAF6 activity. PMID- 14550572 TI - Functional activities and cellular localization of the ezrin, radixin, moesin (ERM) and RING zinc finger domains in MIR. AB - Myosin regulatory light chain interacting protein (MIR) belongs to the ezrin, radixin, moesin (ERM) family of proteins involved in membrane cytoskeleton interactions and cell dynamics. MIR contains, beside the ERM domain, a RING zinc finger region. Immunocytochemistry showed that full-length MIR and the subdomains localize differently in cells. Cell fractionation revealed a similar distribution of full-length MIR and the RING domain protein in the Triton X-100-insoluble fraction. The neurite outgrowth inhibitory activity of MIR was attributed to the RING domain. MIR levels were controlled in the cells depending on the intact RING domain and proteasome activity. The dynamic regulation of MIR contributes to its effects on neurite outgrowth and cell motility. PMID- 14550573 TI - Human immunodeficiency virus-1 Tat protein interacts with distinct proteasomal alpha and beta subunits. AB - The human immunodeficiency virus-1 (HIV-1) Tat protein was previously reported to compete the association of PA28 regulator with the alpha rings of the 20S proteasome and to inhibit its peptidase activity. However, the distinct interaction sites within the proteasome complex remained to be determined. Here we show that HIV-1 Tat binds to alpha4 and alpha7, six beta subunits of the constitutive 20S proteasome and the interferon-gamma-inducible subunits beta2i and beta5i. A Tat-proteasome interaction can also be demonstrated in vivo and leads to inhibition of proteasomal activity. This indicates that Tat can modulate or interfere with cellular proteasome function by specific interaction with distinct proteasomal subunits. PMID- 14550574 TI - Mitochondria from ejaculated human spermatozoa do not synthesize proteins. AB - Sperm motility is dependent on mitochondrial ATP production that relies on the coordinated expression of the mitochondrial and nuclear genomes. It is generally accepted that mammalian ejaculated spermatozoa retain the ability to synthesize mtDNA-encoded proteins but not most of the nuclear ones. This implies an asynchronous regulation of the oxidative phosphorylation-related genes encoded by each genome. Trying to investigate this issue, we unexpectedly found that ejaculated human spermatozoa do not synthesize mtDNA-encoded proteins. Moreover, we estimated that the discrepancy between our observations and those published elsewhere was due to a chloramphenicol-sensitive protein synthesis attributed to mitochondria that instead corresponds to contaminating bacteria. PMID- 14550575 TI - Sodium channel modulating activity in a delta-conotoxin from an Indian marine snail. AB - A 26 residue peptide (Am 2766) with the sequence CKQAGESCDIFSQNCCVG-TCAFICIE NH(2) has been isolated and purified from the venom of the molluscivorous snail, Conus amadis, collected off the southeastern coast of India. Chemical modification and mass spectrometric studies establish that Am 2766 has three disulfide bridges. C-terminal amidation has been demonstrated by mass measurements on the C-terminal fragments obtained by proteolysis. Sequence alignments establish that Am 2766 belongs to the delta-conotoxin family. Am 2766 inhibits the decay of the sodium current in brain rNav1.2a voltage-gated Na(+) channel, stably expressed in Chinese hamster ovary cells. Unlike delta-conotoxins have previously been isolated from molluscivorous snails, Am 2766 inhibits inactivation of mammalian sodium channels. PMID- 14550576 TI - Conditional expression of MCM7 increases tumor growth without altering DNA replication activity. AB - The minichromosome maintenance (MCM) 2-7 complex is a putative DNA helicase complex that facilitates the initiation of DNA replication. Here, we generated a cell line MCM7(+/-)/MCM7-FLAG, in which one allele of MCM7 is mutated whereas a tetracycline-repressible promoter could manipulate the expression of exogenous MCM7 protein. Overexpressed MCM7 protein supports efficient DNA replication of Epstein-Barr virus oriP and rapid formation of tumors in nude mice without altering the activity of cellular DNA replication. This system provides a unique setting for studying the function of MCM7 and for screening for potential therapeutics for malignant tumors. PMID- 14550578 TI - Chairmen's introduction. PMID- 14550579 TI - Risk factors and predictors of compliance in depression. AB - Naturalistic studies show that the average length of antidepressant treatment is less than 6 months and that discontinuations are high, especially in primary care depressed patients. Factors leading to treatment discontinuation are multiple and poorly understood. Discontinuations are most frequent during the first month of therapy, and the prevalent reasons vary during the course of therapy. Approximately 25% of patients do not inform their physician about stopping their antidepressant medication. Recent research provides new information that may serve as a guideline for improving compliance. The physician-patient relationship, including the amount of time the physician takes to explain the expected duration of treatment and possible adverse effects, is a key factor. The physician's attitude about the medication is also important. Strategies should be adapted to the stage of treatment, as common reasons for discontinuation change as treatment progresses. PMID- 14550580 TI - Benefits of different drug formulations in psychopharmacology. AB - Adequate dosage forms are essential for achieving successful pharmacotherapy. Innovative dosage forms or delivery systems may direct a drug to its specific site of action, optimize the timing of the drug release, or increase comfort or convenience for the patient. Thus, such innovations may improve efficacy and tolerability and lead to improvements in health-related quality of life. Specialized dosage forms (e.g., depot injections, extended-release formulations) of several psychiatric agents have been extensively used. The latest addition is an orally disintegrating formulation of the antidepressant mirtazapine. This dosage form dissolves rapidly in the mouth and is convenient for the large proportion of patients who have difficulty in swallowing tablets. PMID- 14550581 TI - Compliance: the impact of adverse events and tolerability on the physician's treatment decisions. AB - A hidden factor that can influence patient compliance is the physician's concern about the tolerability and safety of prescribed medication. Patients may complain about medication side effects that are unpleasant but not dangerous, and this in itself may decrease patient compliance. However, physicians' awareness about adverse events may influence the choice of a drug, and patients' adherence to treatment. In the acute phase of treatment there is a concern about the suicide potential of the depressed patient. The tricyclic antidepressants (TCAs) are toxic in overdose, in contrast to the relative safety of the SSRIs, SNRIs and mirtazapine. Safety issues are also a concern during long-term treatment with antidepressants, i.e., during the continuation and maintenance phases of treatment. Long-term effects such as weight gain and sexual dysfunction distinguish different classes of antidepressants. A particular clinical challenge is the situation when antidepressant medication is administered to a patient with a comorbid medical illness treated with other drugs. This occurs frequently in elderly patients, and may result in an increase in adverse events and/or drug drug interactions. Therefore, one strategy to determine the usefulness of a medication is to study its efficacy, tolerability and adverse events in a vulnerable population. Data from a recently completed open study of mirtazapine orally disintegrating tablets in depressed nursing home patients indicate that it was effective and well tolerated. This study used an orally disintegrating tablet formulation (Remeron SolTab), which was well received by both the patients and staff. Data on patient preferences in another open outpatient study looking at patients at least 50 years of age showed that patients also preferred mirtazapine orally disintegrating tablets to conventional tablets, and indicated that they would be more likely to comply with a prescription for an orally disintegrating tablets than conventional tablets. PMID- 14550582 TI - Involvement of RNA2 for systemic infection of Cucumber mosaic virus isolated from lily on zucchini squash. AB - A lily strain of Cucumber mosaic virus (LK-CMV) was not able to systemically infect zucchini squash (Cucurbita pepo), while Fny strain of CMV (Fny-CMV) caused systemic mosaic and stunting symptom at 4 days post-inoculation on the same host species. The pathogenicity of LK-CMV in zucchini squash was investigated by reassortments of genomic RNAs of LK-CMV and Fny-CMV for infection, as well as by pseudorecombinants generated from biologically active transcripts of cDNA clones of LK-CMV and Fny-CMV, respectively. The assessments of pathogenicity for LK-CMV indicated that RNA2 of LK-CMV was responsible for systemic infection in zucchini squash. In the protoplast of zucchini squash, the RNA accumulations of all constructed pseudorecombinants were indistinguishable and LK-CMV replication was slightly lower than that of Fny-CMV, suggesting that the inability of LK-CMV to infect squash plants was responsible for the poor efficiency of virus movement, rather than the reduction of replication function. PMID- 14550583 TI - Enhanced immunogenicity of SIV Gag DNA vaccines encoding chimeric proteins containing a C-terminal segment of Listeriolysin O. AB - We investigated the potential of the C-terminal 59-amino acid segment of Listeriolysin O (LLO) in enhancing immune responses against the SIV Gag antigen in the context of DNA immunization. Genes with codons optimized for mammalian expression were synthesized for the SIVmac239 Gag, a secreted SIV Gag protein with the tissue plasminogen antigen (tPA) signal fused to its N-terminus (tPA/Gag), as well as their corresponding chimeric proteins Gag/LLO and tPA/Gag/LLO containing the C-terminal 59 amino acids of LLO. Analysis of immune responses to these DNA constructs in a Balb/c mouse model showed that the Gag/LLO construct induced higher levels of both CD4 and CD8 T cell responses against SIV Gag, whereas the tPA/Gag construct induced higher levels of CD4 T cell responses. Moreover, immunization with the tPA/Gag/LLO construct further enhanced both CD4 and CD8 T cell responses. DNA constructs encoding secreted Gag proteins (tPA/Gag and tPA/Gag/LLO) were also more effective in eliciting antibody responses against SIV Gag. Our results demonstrate that the C-terminal segment of LLO can be effectively employed to enhance both cellular and humoral immune responses in the context of a DNA vaccine. PMID- 14550584 TI - Differential activation of interferon-inducible genes by hepatitis C virus core protein mediated by the interferon stimulated response element. AB - We previously found that hepatitis C virus (HCV) core protein, which possesses the consensus sequence of genotype 1b, transcriptionally activates the interferon (IFN)-inducible 2'-5'-oligoadenylate synthetase (2'-5'-OAS) gene in human hepatocyte cells. To clarify the mechanism of this activation, we further characterized the core protein as an activator of the 2'-5'-OAS gene. We demonstrated that the activation of the 2'-5'-OAS gene by the core protein is a general phenomenon, regardless of HCV genotype and strain. We showed that the 20 N-terminal amino acids (aa) of the core protein were important to the activation of the 2'-5'-OAS gene, although this N-terminal region did not have any effect on the subcellular localization of the core protein. We demonstrated that the core protein was able to activate all promoters possessing the IFN-stimulated response element (ISRE) examined. However, we found that the level of activation of the 2' 5'-OAS gene promoter possessing a particular variant type of ISRE was significantly higher than that of other IFN-inducible gene promoters. This phenomenon was confirmed using synthetic promoters possessing five repeats of the consensus or a 2'-5'-OAS-type ISRE. In addition, we showed that gene activation induced by the core protein is mediated by the ISRE. These results imply that the core protein prefers a subclass of IFN-inducible genes, the promoters of which possess the 2'-5'-OAS-type ISRE. Accordingly, we found that the IFN-inducible double-stranded RNA-specific adenosine deaminase gene promoter, possessing a 2' 5'-OAS-type ISRE sequence, was also efficiently activated by the core protein. The exact mechanism by which the core protein enhances gene expression was not determined, but we could find no effects of core protein on gene expression and phosphorylation status of the components of the JAK-STAT signaling transduction pathway. PMID- 14550585 TI - Genetic characterization of wild-type measles viruses in Cambodia. AB - Cambodian authorities in collaboration with the World Health Organization (WHO) are implementing a measles control plan in this measles endemic country. Genetic characterization of Cambodian wild-type measles viruses was performed to determine the genotypes involved in outbreaks, and to measure the level of virus circulation in a geographic area just beginning to implement the measles control program. Seventy-two sequences of the C terminus of the nucleoprotein gene of measles virus were obtained from 88 patients. Samples were taken from 35 among 519 outbreaks reported to the Cambodian National Immunization Program between March 2001 and June 2002. The sequences were grouped into 10 lineages which all belonged to genotype D5. The maximum nucleotide divergence was 1.3%. PMID- 14550586 TI - Sequence variation and evolution of Cryphonectria hypovirus 1 (CHV-1) in Europe. AB - Cryphonectria hypovirus 1 (CHV-1) acts as a naturally occurring biological control agent for chestnut blight, a destructive fungal disease of chestnut trees, which has been introduced into Europe in the 1930s. We have determined partial nucleotide and deduced amino acid sequences of the ORF A of 47 CHV-1 isolates collected in Europe over a period of 28 years. Phylogenetic analysis revealed the presence of four groups or single viruses, which showed sequence divergences ranging from 11 to 19%. These results confirm the previous subtype classification based on RFLP markers, with the exception of the two CHV-1 subtypes E and D, which appear to be related closer than anticipated previously. Dates of divergences between CHV-1 subtypes, calculated from nucleotide substitution rates, indicate that the CHV-1 subtypes diverged several hundreds years ago. Our results suggest that the genetic variation among CHV-1 subtypes did not evolve in Europe and support the hypothesis of multiple introductions of CHV-1 into Europe. PMID- 14550587 TI - Intermediate filament integrity is required for Junin virus replication. AB - The role of the cytoskeletal framework in Junin virus (JUNV) replication has already been demonstrated with compounds interfering with the microfilament (MF) and microtubule (MT) networks. In this work, we evaluated the role of intermediate filaments (IF) during JUNV infection. We tested the effect of acrylamide, a compound that selectively disrupts IF, in culture of three different cell types: Vero cells, murine astrocytes and human foreskin fibroblasts. Perturbation of intermediate filaments had an inhibitory effect on JUNV production within a range of acrylamide concentration of 0.5-3mM in a dose dependent manner, without cell viability modification. Recovery experiments showed that viral production was partially increased when medium containing acrylamide was replaced by normal maintenance medium (MM). The adsorption and internalization steps were not affected by IF disruption. The expression of JUNV proteins was highly reduced in the presence of 2mM acrylamide while immunofluorescence staining of IF showed network disruption with the formation of cytoplasmic aggregates containing vimentin or glial fibrillary acidic protein (GFAP). We conclude that the IF network may play a role in the early step of JUNV multiplication, subsequent to virus entry and that its integrity is a necessary condition for the normal replication of JUNV in neural and fibroblast cells as well as in the Vero cell line. PMID- 14550588 TI - Acetic-acid guided visual inspection vs. cytology-based screening for cervical cancer in the Philippines. AB - OBJECTIVES: To compare the validity and acceptability of acetic-acid visualization (VIA), magnified acetic-acid visualization (VIAM), spatula+cotton swab-Papanicolaou (Pap) smear (SS), and cervical brush-Pap smear (CB) in the detection of precursor/early cervical cancer lesions. METHODS: A total of 12992 women aged between 25 and 65 years from 14 Philippine centers were randomly allocated to the four tests. The gold standard was colposcopy with biopsy for positive/suspicious cases. RESULTS: Sensitivity rates [95% confidence intervals (CIs)] were 37 (CI, 26.8-48.5), 34.1 (CI, 24.8-44.8), 14.3 (CI, 6.4-27.8), and 19.1 (CI, 9.2-34.6) for VIA, VIAM, SS, and CB, respectively. Specificity rates were 90.7 (CI, 89.6-91.7), 90.7 (CI, 89-91.1), 97.5 (CI, 96.8-98), and 97.9 (CI, 97.3-98.4), respectively. Kappa for the Pap smear (PS) within centers ranged from -0.154 to 0.783, and between centers from -0.028 to 0.364. Screeners preferred CB; screened-women preferred VIA. CONCLUSIONS: The acetic-acid visualization and VIAM methods are recommended for initial cervical cancer screening in the Philippines. PMID- 14550589 TI - Human chorionic gonadotropin in cervicovaginal secretions as a predictor of preterm delivery. AB - OBJECTIVES: To determine whether human chorionic gonadotropin (hCG) detected in cervicovaginal secretions of patients with symptoms suggestive of preterm labor is a predictor of preterm birth. METHODS: Women presenting to labor and delivery with symptoms of preterm labor were invited to participate. After collecting secretions with a cotton-tipped applicator from the posterior fornix and endocervical canal, we placed the swab in a test tube containing 1 ml of normal saline. From this solution, we performed qualitative and quantitative hCG measurements. Qualitative and quantitative hCG results from women who delivered at term were compared with those delivering prior to 37 weeks' gestation. RESULTS: Samples were obtained and analyzed from 86 women at a mean (+/- standard deviation) gestational age of 30.7+/-2.9 weeks. Of the 86 subjects enrolled, 28 (32.6%) delivered preterm. A positive rapid qualitative assay from cervicovaginal secretions between 24 and 34 weeks' gestation was associated with a significant increase in the incidence of preterm delivery (relative risk 2.62, 95% confidence interval (CI) 1.38, 4.98). The likelihood ratios for a positive and negative test were 2.19 (95% CI 1.35, 3.56) and 0.51 (95% CI 0.30, 0.85) respectively. Using a cut-off of 19 mIU/ml, similar diagnostic accuracy for predicting preterm birth was obtained with the quantitative test 1.93 (95% CI 1.14, 3.26) and 0.62 (95% CI 0.40, 0.98). CONCLUSIONS: Qualitative and quantitative hCG measurements from cervicovaginal secretions may be useful predictors of preterm birth in symptomatic patients. The qualitative hCG test can be used at the bedside with results obtained in minutes. PMID- 14550590 TI - Anemia during pregnancy in a Chinese population. AB - OBJECTIVES: To study prevalence, risk factors, and birth outcomes of women with anemia during pregnancy in a Chinese population. METHODS: A retrospective cohort study was performed based on 16936 pregnancies delivered between January 1989 and December 1990 in Suzhou, China. Anemia was defined as hemoglobin less than 10 g/dl. Multivariate logistic regression was used to estimate odds ratio, 95% confidence interval, and to control for confounding variables. RESULTS: The prevalence of anemia was 10.3% at the first trimester, 18.9% at the third trimester. Overall, 26.2% pregnant women experienced anemia in pregnancy. Anemia during early pregnancy was not associated with increased risk of adverse perinatal outcomes. However, anemia in later pregnancy was inversely associated with preterm birth and low birth weight. CONCLUSIONS: Anemia during pregnancy does not increase the risk of poor birth outcomes. PMID- 14550592 TI - Transabdominal antepartum amnioinfusion. AB - OBJECTIVES: The purpose of our study was to evaluate whether transabdominal antepartum amnioinfusion is associated with maternal complications during pregnancy and at childbirth. METHODS: Fifty-three pregnant women, hospitalized for oligohydramnios and submitted to transabdominal antepartum amnioinfusion between 16 and 34 weeks' gestational age, were compared with a historic group of 42 pregnant women treated conservatively. RESULTS: The study population was divided into two groups of women with ruptured and unruptured membranes (Groups A and B, respectively). The latency period between time of admission and term of pregnancy was more favorable in amnioinfused patients (Group A: 22 vs. 11 days; Group B: 30 vs. 9 days), and none of the maternal adverse events under study was significantly more common in amnioinfused patients. By contrast, maternal temperature over 38 degrees C was more frequent among controls than among amnioinfused patients with ruptured membranes (23% vs. 4%); so was the number of cesarean sections for fetal distress in Group B (50 vs. 11%). CONCLUSIONS: Antepartum amnioinfusion does not appear to induce greater complications than conservative treatment for oligohydramnios, with or without premature rupture of membranes. On the contrary, this procedure seems to offer several benefits to pregnant women. PMID- 14550591 TI - Intrapartum chlorhexidine vaginal irrigation and chorioamnion and placental microbial colonization. AB - OBJECTIVES: To determine whether intrapartum chlorhexidine vaginal irrigation reduces microbial colonization of the chorioamnion or placenta. METHODS: Secondary analysis was made of a randomized trial. Cultures for aerobic and anaerobic bacteria, Mycoplasma species and Ureaplasma urealyticum were performed using standard isolation techniques. RESULTS: The placentas of 83 trial participants allocated to chlorhexidine and 93 allocated to placebo underwent evaluation. These two groups were statistically balanced for risk factors for infection. Aerobic bacteria were isolated from 47% of the chlorhexidine placentas vs. 51% of the placebo placentas (relative risk 0.9, 95% confidence interval 0.7 1.3), anaerobic bacteria from 30% and 35%, respectively (0.8, 0.5-1.3), group B streptococcus from 12% and 15% (0.8, 0.4-1.7), U. urealyticum from 18% and 29% (0.6, 0.4-1.1), Mycoplasma species from 6% and 11% (0.6, 0.2-1.6), and any organism from 57% and 67%, respectively (0.8, 0.7-1.1). CONCLUSIONS: Intrapartum chlorhexidine vaginal irrigation was associated with non-significant reductions in the rates of placental microbial isolation. PMID- 14550593 TI - Recurrent pregnancy loss due to familial and non-familial habitual molar pregnancy. AB - OBJECTIVES: To present a series of women with recurrent molar pregnancies, including rare familial cases, and discuss etiology and treatment options. METHODS: We performed a detailed clinical evaluation and pedigree analysis of five Egyptian women with recurrent pregnancy loss due to molar pregnancy. RESULTS: The women had a history of four to nine consecutive hydatidiform moles but of no viable pregnancies. Two of the women had molar pregnancies with different husbands who themselves had viable offspring from previous wives; and three of them, who belonged to a family with extensive intermarriage, had a pedigree consistent with an autosomal recessive maternal-effect mutation. CONCLUSIONS: Recurrent pregnancy loss due to habitual molar pregnancy is uncommon and familial cases are extremely rare. The etiology of this disorder is not well understood but likely results from a maternal-effect mutation. Management options are limited, especially for couples who desire to have their own genetic offspring. PMID- 14550594 TI - Thyroid function in hyperemesis gravidarum and correlation with serum leptin levels. PMID- 14550595 TI - Maternal death associated with Eisenmenger's syndrome complicated with HELLP syndrome. PMID- 14550596 TI - Bacterial vaginosis and pregnancy outcome. PMID- 14550597 TI - Peripartum lower urinary tract injury with all-silicone Foley catheters. PMID- 14550598 TI - Preconceptional immunity to cytomegalovirus and the risk of symptomatic congenital infection. PMID- 14550599 TI - Expression of hepatocyte growth factor and its receptor in the placental basal plate in pre-eclamptic pregnancies. PMID- 14550600 TI - Surgical management of tubal prolapse. PMID- 14550601 TI - Cervical adenocarcinoma treated with docetaxel and carboplatin. PMID- 14550602 TI - Preinduction cervical ripening with the Foley catheter and saline infusion vs. cervical dinoprostone. PMID- 14550603 TI - Perceived environmental stress factors and their correlates among pregnant Nigerians. PMID- 14550604 TI - An evaluation of manual vacuum aspiration instruments. AB - OBJECTIVES: Manual vacuum aspiration (MVA) instruments vary in quality, cost, and availability. This evaluation of MVA instruments was conducted to provide decision-makers with information to assist them in determining which instruments to purchase for their specific health care settings. METHODS: Nine MVA devices were evaluated for durability, quality, safety, and usability. Study data came from device specifications, laboratory-based technical studies, and field evaluations. RESULTS: Equipment durability is compromised by high temperatures, quality is compromised by manufacturing defects, and design is key to safe reuse. All MVA instructions are written at advanced reading levels, and device assembly was difficult for most study participants. CONCLUSIONS: No single brand of MVA instrument is perfectly suited for every setting. Users can evaluate the appropriateness of MVA instrument characteristics by reviewing the elements assessed in this evaluation, which, by complementing the experience of administrators and clinicians, should enable them to determine which MVA is best for their environment. PMID- 14550605 TI - A case of sudden death from primary intracranial germinoma complicated by microvascular disease of the heart. AB - A coroner's autopsy conducted on an 11.5-year-old girl, with a history of hypopituitarism, manifesting as growth hormone deficiency associated with short stature, demonstrated the presence of an undiagnosed hypothalamic intracranial germinoma, showing hypophyseal and mesencephalic invasion, together with histological evidence of microvascular (small coronary artery) disease of the heart. The substantive, or primary, cause of her sudden death was clearly the intracranial germinoma, which was acutely haemorrhagic and necrotic. However, it is postulated that there might be an, as yet, undescribed association between this malignancy and the pathogenesis of the coronary microvascular disease; the latter being deemed, in this instance, to be a contributory cause of death. PMID- 14550606 TI - Solid phase micro extraction coupled with on-column GC/ECD for the post-blast analysis of organic explosives. AB - Gas chromatographic analysis with electron capture detection is very sensitive to post-blast residues and useful for the determination of organic explosive molecules. But many compounds extracted from the matrices may interfere with the explosives. Using SPME, most interfering compounds are eliminated so the identification is easier. Another advantage of the technique is a low limit of detection. In this study, four different SPME fibers were tested to analyze the most common encountered organic explosives including nitro aromatics, nitramines and nitro-esters. Different parameters were tested (desorption time, agitation, ...) and a special device has been created to optimize the agitation. Direct desorption effect of the SPME fiber on the column compared to normal split splitless injection is shown. In this way, the degradation of the most sensitive molecules is decreased. An application to a real case is also described in this paper. PMID- 14550607 TI - Acute fatal poisoning with cyamemazine. AB - A case of fatal poisoning with cyamemazine is presented. The cyamemazine was identified in post-mortem blood using a specific gas chromatographic/mass spectrometry method. The autopsy blood concentration of cyamemazine was 1800 ng/ml. Chronic use of cyamemazine was demonstrated by the presence of the drug in hair. Two other drugs were also detected (bromazepam and trimeprazine). We think that this current blood concentration (1800 ng/ml) is a fatal blood concentration because of the negativity of the other parameters, but careful interpretation of analytical findings are important, the possibility that this death was a consequence of the toxicity of combined drugs could not be excluded. Not many therapeutics and toxic levels were previously reported in overdosage cases in which cyamemazine was involved. We consider that this concentration is only of guidance value for a fatal cyamemazine poisoning. PMID- 14550608 TI - Was elusive carnivore a panther? DNA typing of faeces reveals the mystery. AB - In this study, we report the findings of a recent case in which the officials of an Indian zoo claimed that an animal, possibly a carnivore, is periodically visiting the zoo from a nearby vast forest area and causing panic in zoo and nearby villages. They collected some elusive faecal material from the vicinity of an herbivore enclosure. Looking to the pugmarks found in that area and faecal matter ceased, the officials could not decide whether it was a lioness, a tiger or a panther. We resolved this mystery by DNA-based analysis of the faecal material, using our recently developed novel universal primers to amplify and sequence a specific fragment of mitochondrial cytochrome b gene. The findings of the DNA-based analyses were confirmed after few days when the zoo officials trapped the animal of same species as suggested in our report. The potential of our procedure to investigate the cases related to wildlife offence is discussed. PMID- 14550609 TI - The importance of a urine sample in persons intoxicated with flunitrazepam--legal issues in a forensic psychiatric case study of a serial murderer. AB - The sedative-hypnotic benzodiazepine flunitrazepam (FZ) is abused worldwide. The purpose of our study was to investigate violence and anterograde amnesia following intoxication with FZ, and how this was legally evaluated in forensic psychiatric investigations with the objective of drawing some conclusions about the importance of urine sample in a case of a suspected intoxication with FZ. The case was a 23-year-old male university student who, intoxicated with FZ (and possibly with other substances such as diazepam, amphetamines or cannabis), first stabbed an acquaintance and, 2 years later, two friends to death. The police investigation files, including video-typed interviews, the forensic psychiatric files, and also results from the forensic autopsy of the victims, were compared with the information obtained from the case. Only partial recovery from anterograde amnesia was shown during a period of several months. Some important new information is contained in this case report: a forensic analysis of blood sample instead of a urine sample, might lead to confusion during police investigation and forensic psychiatric assessment (FPA) of an FZ abuser, and in consequence wrong legal decisions. FZ, alone or combined with other substances, induces severe violence and is followed by anterograde amnesia. All cases of bizarre, unexpected aggression followed by anterograde amnesia should be assessed for abuse of FZ. A urine sample is needed in case of suspected FZ intoxication. The police need to be more aware of these issues, and they must recognise that they play a crucial role in an assessment procedure. Declaring FZ an illegal drug is strongly recommended. PMID- 14550610 TI - Use of laser microdissection greatly improves the recovery of DNA from sperm on microscope slides. AB - Traditionally, sperms are isolated from vaginal cell mixtures by preferential extraction methods. Although these methods work well when there is a reasonable amount of DNA present, they are problematic when there are limited amounts (ca. 250 pg). In particular, the analysis of sperm from microscope slides has proven difficult. Here, we describe the use of laser capture microdissection (LM) for the isolation of spermatozoa from microscope slides containing sperms and vaginal cells. Such slides are frequently an important source of evidential material during the forensic investigation of rape and other sexual assaults. Low copy number (LCN) PCR was used to compare profiles of sperm DNA prepared using LM and preferential lysis. LM was found to outperform preferential lysis in 15 out of 16 samples. The application of LM to the processing of actual casework slides, and in particular the potential use of LM for the analysis of old cases, is discussed. Finally, 77 post-coital slides were processed in order to accurately assess the robustness of the technique. There was a significant association between the quality of the male profile recovered and time since intercourse that was independent of the number of sperms analysed, suggesting that the DNA was degraded even though the spermhead was intact. PMID- 14550611 TI - Fatal falls from heights in and around Diyarbakir, Turkey. AB - Falls from high places, such as from a building, are frequently encountered in suicides, in some accidents, and sometimes in homicides. In this study, we evaluated the demographic data, mortality rates, fall causes, and post-mortem findings of individuals who fell from heights. Our cases were collected retrospectively from the files of the Branch of the Council of Forensic Medicine in Diyarbakir between 1996 and 2001. There were 431 accidental and 53 suicidal deaths due to blunt injury resulting from falls. Of the victims, 188 were female and 296 were male. The average age of the 484 victims was 27.05 years (range: 4 months-100 years). For buildings, the height ranged from 3 to 8 stories for suicides and from 1 to 8 stories for accidents. We proceeded to analyse the characteristics of accidental falls as follows. The majority of falls were from balconies or rooftops due to the tendency of people to sit and sleep on these places during the hotter months of the year. Some 54.5% of all falls occurred in May-August. The 53 suicidal jumps all occurred off buildings. The victims ranged in age from 15 to 70 years, and comprised 29 women and 24 men. One of the deceased jumped from the roof of a school in which he was boarding, and another from the seventh story of a hospital in which he was receiving treatment. The remaining 51 jumped from heights ranging from 3 to 8 stories. Psychiatric illness was reported in 18 (33.9%) of the suicide deaths, while 10 (18.8%) of the 53 suicides were single women. The results of this study were at variance with literature data with respect to the following: falls from heights were most common in the 0-5 year age group, females had a higher suicide rate than males, and the majority of accidental falls occurred at home rather than in the workplace. PMID- 14550612 TI - Accidental autoerotic deaths between 1978 and 1997. Institute of Legal Medicine, Medical School Hannover. AB - Between 1978 and 1997 the Institute of Legal Medicine of the Hannover Medical School examined 17 fatal autoerotic deaths. The incidence for the Hannover region was 0.49 cases per million inhabitants per year. The victims included 17 men with an average age of 36.8 years; a peak in the age distribution was seen between 20 and 29 years. Twelve of the men were found by friends or family in a domestic environment, while other situations in which the victims were found included the victim's own car, a hotel room, a canal embankment, a public parking lot as well as the holding cell of the youth detention center. The men were of varying socioeconomic status and held a number of different types of jobs or still attended school. Five of the men were found completely nude, while five were only undressed below the waist. Four men wore women's clothes and two were fully clothed with exposed genitals. Besides women's clothes, other objects found at the scene included various types of sexual aids, including ropes, chains, metal bars, locks, sex magazines, condoms, plastic bags, rubber items, etc. In four cases blood alcohol levels between 0.1 and 2.5 per thousand (urine alcohol levels between 0.2 and 2.5 per thousand ) were found. Toxicologic examination revealed chloroform, ketamine, a propane-butane gas mixture in one case each, and in two cases cocaine and morphine. Causes of death included central paralysis after strangulation (seven cases), asphyxiation (4), subarachnoid hemorrhage (2), intoxication (1), hypothermia (1), left heart failure (1), and drowning (1). The history, findings at scene, and autopsy findings and, in individual cases, other investigations are of utmost importance to accurately reconstruct a fatal autoerotic accident. PMID- 14550613 TI - Lethal head injury due to tear-gas cartridge gunshots. AB - We report a fatal head injury caused by a tear-gas cartridge and point out the underestimated potential injury of this type of weapon. Gas pressure wounds and the death mechanisms are also described. A review of the literature and forensic considerations of head injury without projectile are presented. PMID- 14550614 TI - Human pentastomiasis discovered postmortem. AB - The autopsy of an 18-year-old girl who had died suddenly at home revealed generalised pentastomiasis. The location of this disease in the intestines was responsible for death by hemorrhagic enterocolitis. This discovery constituted an atypical case in our daily practice of forensic medicine. PMID- 14550615 TI - Diagnosing death by drowning in fresh water using blood strontium as an indicator. AB - The interpretation of the level of strontium (Sr) in blood has been demonstrated to be useful in the diagnosis of death by drowning occurred in sea water, but its use in fresh water drownings is not so evident because of the low Sr concentration present in most of fresh water media. In this paper, we show a survey of the results obtained in the casework analysis of Sr in ventricular blood used in the diagnosis of drowning in 144 bodies found immersed in fresh water over a period of 10 years. Thirty-two percent of the immersion cases examined could be diagnosed as drownings with a reasonable degree of confidence. It is thought that this percentage of positive diagnoses could be largely improved in the case of blood samples taken a few hours after death. PMID- 14550616 TI - Homicidal and suicidal ligature strangulation--a comparison of the post-mortem findings. AB - Suicides by ligature strangulation are rare events. In Berlin (3.5 million inhabitants; ca. 500 suicides per year) approximately one case per year occurs. Here, we present the main findings of 19 cases investigated between 1978 and 1998, compared to 47 cases of homicidal ligature strangulation. Two of the 19 suicidal victims had single fractures of the upper thyroid horns and one victim a fracture of a lower thyroid horn; other types of laryngohyoid injuries were not observed. In the homicidal series, the laryngohyoid structures were unaffected in 26 cases (12 of these victims were children or adolescents), single thyroid horn fractures were present in three cases and more significant injuries in 18 cases. Macroscopic bleedings of the laryngeal muscles were found in 12 victims of the homicidal group and in none of the suicidal. Bleedings in the neck muscles seldom occurred in suicides. According to these findings, the laryngohyoid injuries can be helpful in the differentiation of suicide from homicide, if more than a single thyroid horn fracture or a laryngeal soft tissue trauma is present. PMID- 14550617 TI - Analysis of the CODIS autosomal STR loci in four main Colombian regions. AB - Genotype polymorphism studies at the 13 loci STRs included in the combined DNA index system [CODIS and PCR-based short tandem repeat loci, in: Proceedings of the Second European Symposium on Human Identification, Promega Corporation, Madison, WI, 1998, pp. 73-88; J. Forensic Sci. 46 (2001) 453] (CODIS: D3S1358, HUMvWA31, HUMFGA, D8S1179 D21S11, D18S51, D5S818, D13S317, D7S820, HUMTH01, HUMTPOX, HUMCSF1PO and D16S539) were carried out in a sample of 1429 unrelated Colombian individuals belonging to 25 different departments. As many other countries in Latino-America, Colombia shows an important admixture component, basically integrated by Amerindians, European-descendants and African descendants. Due to the fact that only partial population analyses have been carried out in the country, the main aim of the present analysis is to establish a database of forensic interest based on the widely used CODIS systems covering the main Colombian regions. PMID- 14550618 TI - Multiplex PCR and minisequencing of SNPs--a model with 35 Y chromosome SNPs. AB - We have developed a robust single nucleotide polymorphism (SNPs) typing assay with co-amplification of 25 DNA-fragments and the detection of 35 human Y chromosome SNPs. The sizes of the PCR products ranged from 79 to 186 base pairs. PCR primers were designed to have a theoretical Tm of 60 +/- 5 degrees C at a salt concentration of 180 mM. The sizes of the primers ranged from 19 to 34 nucleotides. The concentration of amplification primers was adjusted to obtain balanced amounts of PCR products in 8mM MgCl2. For routine purposes, 1 ng of genomic DNA was amplified and the lower limit was approximately 100 pg DNA. The minisequencing reactions were performed simultaneously for all 35 SNPs with fluorescently labelled dideoxynucleotides. The size of the minisequencing primers ranged from 19 to 106 nucleotides. The minisequencing reactions were analysed by capillary electrophoresis and multicolour fluorescence detection. Female DNA did not influence the results of Y chromosome SNP typing when added in concentrations more than 300 times the concentrations of male DNA. The frequencies of the 35 SNPs were determined in 194 male Danes. The gene diversity of the SNPs ranged from 0.01 to 0.5. PMID- 14550619 TI - Haplotypes from the Caucasus, Turkey and Iran for nine Y-STR loci. AB - Nine Y chromosome short tandem repeat (STR) loci (DYS385a, DYS385b, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393 and DYS19 (DYS394)) were typed in 669 individuals belonging to 16 populations from the Caucasus, Turkey and Iran. PMID- 14550620 TI - Genetic diversity and forensic evaluation of 10 STR loci in Lodz region of Poland. AB - Allele frequency data and forensic efficiency parameters for 10 STR loci: D3S1358, vWA, D16S539, D2S1338, D8S1179, D21S11, D18S51, D19S433, TH01 and FGA, were estimated from a sample of 207 unrelated individuals in Lodz region of Poland. PMID- 14550621 TI - Population genetics of nine STR loci: TH01, TPOX, CSF1PO, vWA, FESFPS, F13A01, D13S317, D7S820 and D16S539 in a Korean population. AB - Allele frequencies for nine STR loci namely, TH01, TPOX, CSF1PO, vWA, FESFPS, F13A01, D13S317, D7S820 and D16S539 were obtained from a sample of 437 unrelated individuals living in Chungcheong-do, South Korea. PMID- 14550622 TI - Polymorphism of LDLR, GYPA, HBGG, D7S8, GC, HLA-DQA1, Ig-JH, D17S30, ApoB and D1S80 loci in northwestern Russians. AB - Allele frequencies for polymarker, HLA-DQA1, Ig-JH, D17S30, ApoB and D1S80 loci and population genetic parameters were obtained from a sample of 501 unrelated individuals born in the northwestern Federal Region of Russia. PMID- 14550623 TI - STR-CODIS typing in Greece. AB - Thirteen STRs loci have been typed in a sample from Greece. PMID- 14550624 TI - Will fatty worms help cure human obesity? AB - The incidence of obesity has reached epidemic proportions within industrial societies; however, research on human obesity has been hampered by our inability to control genetic and environmental factors. The control of energy homeostasis appears to be conserved among species. Recent creative research in Caenorhabditis elegans, including the application of a genome-wide RNA interference analysis, has provided insight into the genes involved in energy balance. In this article, we discuss the results of these studies and their potential importance to humans. PMID- 14550625 TI - Chloroplast-to-nucleus signalling: a role for Mg-protoporphyrin. AB - Signalling from chloroplasts to the nucleus is an important feature of the coordination of nuclear and chloroplast gene expression required for the assembly of functional chloroplasts. Recent studies have indicated that accumulation of Mg protoporphyrin, the first committed precursor of chlorophyll, is both necessary and sufficient for the regulation of nuclear genes by chloroplasts. PMID- 14550626 TI - Transposable elements in mammals promote regulatory variation and diversification of genes with specialized functions. AB - Nearly half of mammalian genomes are derived from ancient transposable elements (TEs). We analyzed the prevalence of TEs in untranslated regions of human and mouse mRNAs and found evidence suggesting that TEs affect the expression of many genes through the donation of transcriptional regulatory signals. Furthermore, we found that recently expanded gene classes, such as those involved in immunity or response to external stimuli, have transcripts enriched in TEs, whereas TEs are excluded from mRNAs of highly conserved genes with basic functions in development or metabolism. These results support the view that TEs have played a significant role in the diversification and evolution of mammalian genes. PMID- 14550627 TI - False discoveries and models for gene discovery. AB - In the search for genes underlying complex traits, there is a tendency to impose increasingly stringent criteria to avoid false discoveries. These stringent criteria make it hard to find true effects, and we argue that it might be better to optimize our procedures for eliminating and controlling false discoveries. Focusing on achieving an acceptable ratio of true- and false-positives, we show that false discoveries could be eliminated much more efficiently using a stepwise approach. To avoid a relatively high false discovery rate, corrections for 'multiple testing' might also be needed in candidate gene studies. If the appropriate methods are used, detecting the proportion of true effects appears to be a more important determinant of the genotyping burden than the desired false discovery rate. This raises the question of whether current models for gene discovery are shaped excessively by a fear of false discoveries. PMID- 14550628 TI - Running with RNA polymerase: eukaryotic transcript elongation. AB - Long recognized as a target of regulation in prokaryotes, transcript elongation has recently become the focus of many investigators interested in eukaryotic gene expression. The growth of this area has been fueled by the availability of new methods and molecular structures, expanding sequence databases and an appreciation for the exquisite coordination required among different processes in the nucleus. Our article collates new information on regulatory accessory factors, as well as their ultimate target, RNA polymerase, in the nucleus of eukaryotic cells. How this regulation influences the biology of the organism is quite profound, and from single cell to multicellular eukaryotes significant similarities exist in the molecular responses to extracellular signals during transcript elongation. The most advanced genetic knowledge in this area comes from Saccharomyces cerevisiae, but the biochemistry and cell biology results from other organisms are also highlighted. PMID- 14550629 TI - Integrating 'omic' information: a bridge between genomics and systems biology. AB - The availability of genome sequences for several organisms, including humans, and the resulting first-approximation lists of genes, have allowed a transition from molecular biology to 'modular biology'. In modular biology, biological processes of interest, or modules, are studied as complex systems of functionally interacting macromolecules. Functional genomic and proteomic ('omic') approaches can be helpful to accelerate the identification of the genes and gene products involved in particular modules, and to describe the functional relationships between them. However, the data emerging from individual omic approaches should be viewed with caution because of the occurrence of false-negative and false positive results and because single annotations are not sufficient for an understanding of gene function. To increase the reliability of gene function annotation, multiple independent datasets need to be integrated. Here, we review the recent development of strategies for such integration and we argue that these will be important for a systems approach to modular biology. PMID- 14550630 TI - The enigma of ribonuclease P evolution. AB - The 5'-end maturation of tRNAs is catalyzed by the ribonucleoprotein enzyme ribonuclease P (RNase P) in all organisms. Here we provide, for the first time, a comprehensive overview on the representation of individual RNase P protein homologs within the Eukarya and Archaea. Most eukaryotes have homologs for all four protein subunits (Pop4, Rpp1, Pop5 and Rpr2) present in the majority of Archaea. Pop4 is the only RNase P protein subunit identifiable in all Eukarya and Archaea with available genome sequences. Remarkably, there is no structural homology between bacterial and archaeal-eukaryotic RNase P proteins. The simplest interpretation is that RNase P has an 'RNA-alone' origin and progenitors of Bacteria and Archaea diverged very early in evolution and then pursued completely different strategies in the recruitment of protein subunits during the transition from the 'RNA-alone' to the 'RNA-protein' state of the enzyme. PMID- 14550631 TI - Comparison and meta-analysis of microarray data: from the bench to the computer desk. AB - The upcoming availability of public microarray repositories and of large compendia of gene expression information opens up a new realm of possibilities for microarray data analysis. An essential challenge is the efficient integration of microarray data generated by different research groups on different array platforms. This review focuses on the problems associated with this integration, which are: (1) the efficient access to and exchange of microarray data; (2) the validation and comparison of data from different platforms (cDNA and short and long oligonucleotides); and (3) the integrated statistical analysis of multiple data sets. PMID- 14550632 TI - The transcriptional complexity of the TFIIH complex. AB - Mutations in some subunits of the basal DNA repair and transcription factor II H (TFIIH) are involved in several human genetic disorders. Transcription factor II H interacts with a variety of factors during transcription, including nuclear receptors, tissue-specific transcription factors, chromatin remodeling complexes and RNA, suggesting that, in addition to its essential role in transcription initiation, it also participates as a regulatory factor. Interpretation of the phenotypes produced by mutations in TFIIH is complicated by the recent finding that TFIIH plays a role in RNA polymerase I (RNA Pol I)-mediated transcription. In vitro reconstituted systems and genetic analysis suggest two possible explanations for the transcriptional phenotypes of TFIIH mutations that are not mutually excluding. The first is that different sets of genes require different levels of transcription to maintain a wild-type phenotype. The second suggests that mutations in TFIIH produce specific phenotypes arising from differential interactions of this complex with different transcription regulatory factors. PMID- 14550633 TI - Recombinant Yersinia enterocolitica YscM1 and YscM2: homodimer formation and susceptibility to thrombin cleavage. AB - Pathogenic Yersinia species (Y. enterocolitica, Y. pestis, and Y. pseudotuberculosis) make use of a virulence plasmid-encoded type three secretion system (TTSS) to inject effector proteins into host cells. Y. enterocolitica YscM1 (LcrQ in Y. pestis and Y. pseudotuberculosis) and its homologue YscM2 are regulatory components of the TTSS that are also secreted by this transport apparatus. YscM1 and YscM2 share 57% identity and are believed to be functionally equivalent. We have recombinantly expressed and purified YscM1 and YscM2 in Escherichia coli. After expression as glutathione S-transferase (GST) fusions purification to near homogeneity was achieved by glutathione-Sepharose affinity chromatography followed by PreScission protease treatment to cleave off GST and gel filtration on a Superdex 75 column. Such recombinant YscM1 and YscM2 bound efficiently to the specific chaperone SycH, indicating proper folding of the purified proteins. Gel filtration analyses revealed that both YscM1 and YscM2 formed homodimers. The YscM1 and YscM2 homodimers could be dissociated at high ionic strength, indicating that salt bridges essentially contribute to the dimerization. We further demonstrated that YscM1 and YscM2 are susceptible to thrombin cleavage. PMID- 14550634 TI - Overexpression of juvenile hormone binding protein in bacteria and Pichia pastoris. AB - Galleria mellonella juvenile hormone binding protein (JHBP) is a single chain glycoprotein with two disulfide bonds and a molecular mass of 25,880 Da. This report describes the expression of JHBP in bacteria and yeast cells (Pichia pastoris). The expression in bacteria was low and the protein was rapidly degraded upon cell lysis. The expression of His8-tagged rJHBP (His8-rJHBP) in P. pastoris was high and the non-degraded protein was purified to homogeneity with high yield in a one-step immobilized Ni++ affinity chromatography. His8-rJHBP from P. pastoris contains one JH III binding site with KD of 3.7 +/- 1.3x10(-7) M. The results suggest that P. pastoris is the preferred system for expression of His8-rJHBP in non-degraded fully active form. PMID- 14550635 TI - Chromatography of isoforms of recombinant apoaequorin and method for the preparation of aequorin. AB - Gradient elution chromatography of recombinant apoaequorin carried out in the presence of Ca2+ revealed two isoforms of apoaequorin, reduced and oxidized, whereas in the presence of EDTA 3 isoforms were observed. In a regeneration mixture of apoaequorin, coelenterazine, EDTA, and 2-mercaptoethanol, four isoforms were obtained, of which only one, aequorin, gave light with Ca2+. A method is described for the preparation of highly pure aequorin. The aequorin was stable in solution for approximately 10 days at 4 degrees C and pH 7.6, and then it gradually lost activity with a half-life of about 20 days until it was almost completely inactive on day 30. PMID- 14550636 TI - Bacterial production and purification of SGPI-1 and SGPI-2, two peptidic serine protease inhibitors from the desert locust, Schistocerca gregaria. AB - The last decade, a new serine protease inhibitor family has been described in arthropods. Eight members were purified from the locusts Locusta migratoria (LMPI 1-2 and HI) and Schistocerca gregaria (SGPI-1-5) and 11 additional locust peptides were identified by cDNA cloning. Furthermore, the light chain of the 155 kDa heterodimeric protease inhibitor pacifastin, from the freshwater crayfish Pacifastacus leniusculus, was found to be composed of nine consecutive inhibitory domains (PLDs). These domains share a pattern of 6 conserved cysteine residues (Cys-Xaa(9-12)-Cys-Asn-Xaa-Cys-Xaa-Cys-Xaa(2-3)-Gly-Xaa(3-4)-Cys-Thr-Xaa3-Cys) with the locust inhibitors. So far, for most of the PLD-related peptides the biological functions remain obscure. To obtain sufficient amounts of material to perform physiological experiments, we have optimised the production of SGPI-1-2 via a bacterial (Escherichia coli) expression system. The cDNA sequences encoding these peptides were inserted in the pMAL-2pX vector, downstream of the gene encoding the maltose-binding protein (including a signal peptide). As a consequence, both peptides were expressed as fusion proteins (2-3 mg/l) and targeted to the periplasmic space. Following a one-step affinity purification, both fusion proteins were successfully cleaved by Factor Xa and after a methanol extraction, it took only one additional RP-HPLC run to purify both peptides to homogeneity. Finally, the formation of the disulphide bridges and the biological activity of the recombinant peptides were verified by mass spectrometry and a spectrophotometric protease inhibitor assay, respectively. PMID- 14550638 TI - Expression and purification of polyhistidine-tagged rotavirus NSP4 proteins in insect cells. AB - The rotavirus nonstructural NSP4 protein, a transmembrane endoplasmic reticulum specific glycoprotein, has been described as the first viral enterotoxin. Purified NSP4 or a peptide corresponding to NSP4 residues 114-135 induces diarrhea in young mice. NSP4 has a membrane-destabilizing activity and causes an increase in intracellular calcium levels and chloride secretion by a calcium dependent signalling pathway in eucaryotic cells. In this study, four recombinant baculoviruses were generated expressing the rotavirus NSP4 glycoprotein from the human strains Wa and Ito, the porcine strain OSU, and the simian strain SA11, which belong to two different NSP4 genotypes, A and B. The recombinant glycoproteins, expressed as polyhistidine-tagged molecules, were analyzed by Western blotting and immunoprecipitation. Newborn mice responded with diarrhea after inoculation with each of the recombinant NSP4 proteins. PMID- 14550637 TI - Expression of a novel regenerating gene product, Reg IV, by high density fermentation in Pichia pastoris: production, purification, and characterization. AB - Human regenerating (Reg) gene products are regionally expressed by gut-derived tissues, and are markedly up-regulated in cancer and in diseases characterized by mucosal injury. We recently identified Reg IV, a novel regenerating gene product that is uniquely expressed by the normal distal gastrointestinal mucosa. The function remains poorly understood due to the lack of significant purified Reg IV for biochemical and functional studies. Recombinant human Reg IV was efficiently expressed under the control of the AOX1 gene promoter in Pichia pastoris using the MutS strain KM71H. We describe the unique conditions that are required for efficient production of Reg IV protein in high density fermentation. Optimal protein expression was obtained by reduction of the fermentation temperature and addition of casamino acids as a supplemental nitrogen source and to minimize the activity of yeast produced proteases. Recombinant Reg IV protein was purified by tangential flow filtration and reverse phase chromatography. The purified protein was characterized by amino terminus sequence analysis and MALDI-TOFMS showing that the engineered protein had the expected sequence and molecular weight without secondary modification. Recombinant Reg IV was further characterized by specific monoclonal and polyclonal reagents that function for Western blot analysis and for immunolocalization studies. PMID- 14550639 TI - Heterologous expression in Pichia pastoris and single-step purification of a cysteine proteinase from northern shrimp. AB - A distinct cysteine proteinase (NsCys) of northern shrimp Pandalus borealis belonging to cathepsin L subgroup of the papain superfamily has been overexpressed as a precursor form (proNsCys) in Pichia pastoris. We adopted a simple and quick procedure to generate an expression cassette by constructing a donor vector harboring proNsCys followed by recombination with an acceptor vector in a way so that the proNsCys gene was placed downstream of the methanol inducible AOX1 promoter and alpha-mating factor signal sequence gene. In addition, we used glycerol complex medium that supported high growth of yeast before induction while induction was carried out in minimal methanol medium thereby facilitating the secreted protein to be purified with a single size exclusion chromatography. The recombinant enzyme was purified in two enzymatically active fractions: both corresponding to mature NsCys with, however, the major one comprising two molecular species of NsCys which had their severed prodomain non-covalently attached. The overall yield was about 100 mg of crude or 60 mg of purified recombinant enzyme comprising both mature and prodomain attached forms of NsCys per liter of yeast culture. The recombinant NsCys was biologically active as observed by gelatin zymography and its ability to cleave Z Phe-Arg-MCA, a synthetic substrate for cathepsin L. The development of the system reported here provides a cost-effective and easy to manipulate expression system to obtain large quantities of fully functional shrimp enzyme that will enable the functional characterization of this unique enzyme for both research and industrial purposes. PMID- 14550640 TI - High yield expression, refolding, and characterization of recombinant interferon alpha2/alpha8 hybrids in Escherichia coli. AB - Interferons (IFNs) are a family of pleiotropic cytokines used for the treatment of various viral infections and cancers. The low-cost production of IFNs with high biological value and the discovery of IFNs with improved properties are important for the treatment of these diseases as well as for understanding the physiological functions of these compounds. We describe a protein expression system for the production of IFNs alpha2, alpha8, and their hybrids in insoluble form in Escherichia coli, coupled to an efficient two-step optimized refolding and histidine-tag purification protocol. The expressed IFNs were of high biological value, as shown in antiviral and antiproliferative assays and some had specific activities higher than those of the commercially available interferon preparations and exhibited novel properties. This time-efficient, optimized protein expression method allows for the production of not just a single interferon subtype but several native and hybrid IFNs with relatively high yield and low cost that can be used in functional and potentially clinical assays. PMID- 14550642 TI - Purification of enzymatically active human lysyl oxidase and lysyl oxidase-like protein from Escherichia coli inclusion bodies. AB - Lysyl oxidase (LOX) is an extracellular copper dependent enzyme catalyzing lysine derived cross-links in extracellular matrix proteins. Recent molecular cloning has revealed the existence of a LOX family consisting of LOX and four lysyl oxidase-like proteins (LOXLs; LOXL, LOXL2, LOXL3, and LOXL4). Each member of the LOX family contains a copper-binding domain, residues for lysyl-tyrosyl quinone, and a cytokine receptor-like domain. Very recently, novel functions, such as tumor suppression, cellular senescence, and chemotaxis, have been attributed to this family of amine oxidases, but functional differences among the family members have yet to be determined. For efficient expression and purification, we cloned the cDNAs corresponding to proteolytically processed forms of LOX (LOX-p) and LOXL (LOXL-p1 and LOXL-p2) into a bacterial expression vector pET21a with six continuous histidine codons attached to the 3' of the gene. The recombinant proteins were purified with nickel-chelating affinity chromatography and converted into enzymatically active forms by stepwise dialysis in the presence of N-lauroylsarcosinate and Cu2+. The purified LOX-p, LOXL-p1, and LOXL-p2 proteins showed specific amine oxidase activity of 0.097, 0.054, and 0.150 U/mg, respectively, which was inhibited by beta-aminopropionitrile (BAPN), a specific inhibitor of LOX. Availability of these pure and active forms of LOX and LOXLs will be significantly helpful in functional studies related to substrate specificity and crystal structures of this family of amine oxidases. PMID- 14550641 TI - Expression and purification of the h1 and h2 isoforms of calponin. AB - Three homologous calponin isoforms, named h1, h2, and acidic calponins, have been found in birds and mammals. Based primarily on studies of chicken gizzard smooth muscle (h1) calponin, calponin has been identified as a family of actin associated proteins that inhibit actomyosin ATPase activity. Evolutionary divergence of the calponin isoforms suggests differentiated function. While the role of h1 calponin in smooth muscle contraction is under investigation, h2 calponin has been shown regulating the function of actin cytoskeleton. Using cloned cDNA, we expressed mammalian h1 and h2 calponins in Escherichia coli. We have developed effective methods to purify biologically active h1 and h2 calponin proteins from transformed bacterial culture. The purified calponin isoform proteins were used to generate monoclonal antibodies that reveal epitopic structure difference between h1 and h2 calponins. Together with their differential expression in tissues and during development, the structural diversity of h1 and h2 calponins suggests non-redundant physiological function. Nevertheless, h1 and h2 calponins bind F-actin with similar affinity, indicating a conserved mechanism for their role in regulating actin filaments in smooth muscle and non-muscle cells. PMID- 14550643 TI - Codon optimizer: a freeware tool for codon optimization. AB - Selection plays a major role in the determination of codon usage in all organisms studied so far. In highly expressed genes, a narrow set of codons is used and these codons correspond to the more abundant tRNA species. This minimizes the risk of tRNA depletion during translation. In fact, the codons in a gene may be true bottlenecks, especially in cases where foreign genes are expressed in a host in which the usage of codons in highly expressed genes does not resemble the usage of codons in the species from which the foreign gene originates. In such cases, it has been shown that substitution of rare codons in the introduced gene may increase the yield dramatically. In addition, replacement of rare codons might decrease the chance of misincorporation and protect the protein from premature turnover. Here, a piece of software is announced that calculates a codon-optimized sequence of any gene based on knowledge of highly expressed genes of a host. In addition, it calculates the codon adaptation index of the gene and identifies internal type II restriction sites of the optimized sequence. The program runs under Windows and is available as freeware for use in academia. PMID- 14550644 TI - High-yield expression in Escherichia coli, purification, and characterization of properly folded major peanut allergen Ara h 2. AB - Allergic reactions to peanuts are a serious health problem because of their high prevalence, associated with potential severity, and chronicity. One of the three major allergens in peanut, Ara h 2, is a member of the conglutin family of seed storage proteins. Ara h 2 shows high sequence homology to proteins of the 2S albumin family. Presently, only very few structural data from allergenic proteins of this family exist. For a detailed understanding of the molecular mechanisms of food-induced allergies and for the development of therapeutic strategies knowledge of the high-resolution three-dimensional structure of allergenic proteins is essential. We report a method for the efficient large-scale preparation of properly folded Ara h 2 for structural studies and report CD spectroscopic data. In contrast to other allergenic 2S albumins, Ara h 2 exists as a single continuous polypeptide chain in peanut seeds, and thus heterologous expression in Escherichia coli was possible. Ara h 2 was expressed as Trx-His-tag fusion protein in E. coli Origami (DE3), a modified E. coli strain with oxidizing cytoplasm which allows the formation of disulfide bridges. It could be shown that recombinant Ara h 2, thus overexpressed and purified, and the allergen isolated from peanuts are identical as judged from immunoblotting, analytical HPLC, and circular dichroism spectra. PMID- 14550645 TI - Purification and characterization of human nucleolar phosphoprotein 140 expressed in Escherichia coli. AB - Human nucleolar phosphoprotein 140, hNopp140, is one of the most highly phosphorylated mammalian proteins, which is involved in the biogenesis of nucleolus. It regulates the transcription of rDNA and has a tendency to bind to doxorubicin, which is widely used as an anti-cancer drug. The biochemical and biophysical property of hNopp140 has not been reported due to the fact that it is rather difficult to obtain protein in large enough quantity. In this paper, we report the cloning and overexpression of the soluble form of hNopp140 in Escherichia coli. The protein was purified to more than 90% homogeneity using hydroxyapatite and ion exchange chromatography. The purified protein can be extensively phosphorylated by casein kinase II and oligomerized into an insoluble aggregate in the presence of magnesium, carbonate, and fluoride ions. PMID- 14550646 TI - Purification and characterization of hexahistidine-tagged elongation factor SelB. AB - The cotranslational incorporation of selenocysteine into proteins is mediated by a specialized elongation factor, named SelB. Its amino-terminal three domains show homology to elongation factor EF-Tu and accordingly bind GTP and selenocysteyl-tRNASec. In addition, SelB exhibits a long carboxy-terminal extension that interacts with a secondary structure of selenoprotein mRNAs (SECIS element) positioned immediately downstream of the in-frame UGA codons specifying the sites of selenocysteine insertion. In this report, a fast and efficient method for the purification of large amounts of hexahistidine-tagged SelB is presented. After two chromatographic steps, 10 mg pure protein was isolated from 12 g wet cell pellet. Biochemical analysis of the purified protein showed that the tag does not influence the interaction of SelB with guanine nucleotides, SECIS elements, and selenocysteyl-tRNASec. In addition, the fusion protein is fully functional in mediating UGA read-through in vivo. It therefore represents an excellent model for studying the function of SelB and the mechanisms of selenocysteine incorporation. PMID- 14550647 TI - Large scale production of biologically active Escherichia coli glutamyl-tRNA reductase from inclusion bodies. AB - Glutamyl-tRNA reductase catalyzes the initial step of tetrapyrrole biosynthesis in plants and prokaryotes. Recombinant Escherichia coli glutamyl-tRNA reductase was purified to apparent homogeneity from an overproducing E. coli strain by a two-step procedure yielding 5.6 mg of enzyme per gram of wet cells with a specific activity of 0.47 micromol min(-1)mg(-1). After recombinant production, denatured glutamyl-tRNA reductase from inclusion bodies was renatured by an on column refolding procedure. Residual protein aggregates were removed using Superdex 200 gel-filtration chromatography. Solubility, specific activity, and long-term storage properties were improved compared to previous protocols. Obtained enzyme amounts of high purity now allow the research on the recognition mechanism of tRNAGlu and high-throughput inhibitor screening. PMID- 14550648 TI - Molecular cloning, overexpression in Escherichia coli, and purification of 6x his tagged C-terminal domain of Clostridium difficile toxins A and B. AB - Genomic DNA from ribotype-01 and -17 Clostridium difficile strains was used for amplification of the sequences encoding the carboxy-terminal domain of toxins A (TcdA) and B (TcdB). The deduced C-terminal TcdB ribotype-01 and -17 domains share 99.5% amino acid sequence identity while TcdA ribotype-17 comprises a 607 amino acid deletion compared to TcdA-01. When compared to previously sequenced C. difficile toxins, 99.3% amino acid identity was found between TcdA-01 and TcdA from strain VPI10643 and 98.8% identity between TcdA-17 and TcdA from strain F 1470. The obtained sequences were fused in 3' to a sequence encoding a hexahistidine tag and cloned into an Escherichia coli expression vector. The recombinant proteins were expressed in E. coli and purified using single-step metal-chelate chromatography. The recombinant carboxy-terminal domain of TcdA-01 was purified from the soluble E. coli lysate fraction whereas TcdA-17 and TcdB-17 carboxy-terminal domains were purified from inclusion bodies. At least 40 mg of each protein was purified per liter of bacterial culture. The recombinant toxin domains were detected specifically by Western blot and ELISA with antibodies against native C. difficile toxins. This study demonstrated that the carboxy terminal domains of TcdA and TcdB can be produced using an E. coli expression system and easily purified. These recombinant, stable, and non-toxic proteins provide a convenient source for use in the diagnosis of C. difficile infections, instead of native toxins, as controls and calibrators in immunoassay kits and to obtain specific monoclonal antibodies. PMID- 14550649 TI - Cloning, expression, and characterization of a bi-functional disintegrin/alkaline phosphatase hybrid protein. AB - Integrins are transmembrane heterodimeric glycoproteins responsible for cellular communication; therefore, they play an essential role in many physiological events. Viper snake venoms contain integrin antagonists called disintegrins which bind and inhibit integrin function. They present a loop containing an RGD motif responsible for integrin binding. The engineering of disintegrins fused to a reporter enzyme will be an interesting approach to build integrin markers. Even more, the disintegrin scaffold could be used to present other protein binding motifs. In this work, we have obtained alkaline phosphatase (APv) tagged eristostatin (Er) by cloning and expressing eristostatin DNA into the pLIP6-GN vector. Eristostatin, a 49 residue disintegrin, binds selectively to alphaIIbbeta3 integrin, inhibiting its binding to fibrinogen. The resulting fusion protein Er/APv was identified by SDS-PAGE and by Western blotting using both anti-Er and anti-AP antibodies. This fusion protein showed enzymatic AP activity similar to that of wild APv and its potential use for an alphaIIbbeta3 integrin assay was tested in a one-step dot blot using immobilized cells incubated with the marker and developed by AP substrate. Er/APv showed selectivity towards platelets and alphaIIbbeta3 integrin transfected cells and reacted with the same region as unlabeled Er, as analyzed in competition assays. Our data present a novel tool, Er/APv, with potential use as molecular marker in processes where the alphaIIbbeta3 integrin is involved. PMID- 14550650 TI - Expression, purification, and characterization of His-tagged human mitochondrial 2,4-dienoyl-CoA reductase. AB - Mitochondrial 2,4-dienoyl-CoA reductase is a key enzyme for the beta-oxidation of unsaturated fatty acids. The cDNA of the full-length human mitochondrial 2,4 dienoyl-CoA reductase was previously cloned as pUC18::DECR. PCR methodologies were used to subclone the genes encoding various truncated human mitochondrial 2,4-dienoyl-CoA reductases from pUC18::DECR with primers that were designed to add six continuous histidine codons to the 3' or 5' primer. The PCR products were inserted into pLM1 expression vectors and overexpressed in Escherichia coli. A highly active truncated soluble protein was expressed and purified with a nickel HiTrap chelating metal affinity column to apparent homogeneity based on Coomassie blue-stained SDS-PAGE. The molecular weight of the protein subunit was 34 kDa. The purified protein is highly stable at room temperature, which makes it potentially valuable for protein crystallization. KM of 26.5 +/- 3.8 microM for 2,4-hexadienoyl-CoA, KM of 6.22 +/- 2.0 microM for 2,4-decadienoyl-CoA, and KM of 60.5 +/- 19.7 microM for NADPH, as well as Vmax of 7.78 +/- 1.08 micromol/min/mg for 2,4-hexadienoyl-CoA and Vmax of 0.74 +/- 0.07 micromol/min/mg for 2,4 decadienoyl-CoA were determined on kinetic study of the purified protein. The one step purification of the highly active human mitochondrial 2,4-dienoyl-CoA reductase will greatly facilitate further investigation of this enzyme through site-directed mutagenesis and enzyme catalyzed reactions with substrate analogs as well as protein crystallization for solving its three-dimensional structure. PMID- 14550651 TI - Expression, purification, and characterization of a bacterial GTP-dependent PEP carboxykinase. AB - The Corynebacterium glutamicum (C. glutamicum) phosphoenolpyruvate carboxykinase (PCK) gene (pckA) was cloned into an Escherichia coli expression vector with a glutathione S-transferase (GST) tag. This recombinant DNA can produce highly overexpressed tagged protein in soluble form. This is the first report of the production of C. glutamicum PCK overexpressed in E. coli. The GST-fused PCK was purified using the glutathione-Sepharose 4B affinity column and the GST tag was removed in one-step. This one-step, easy purification method would be very useful for future mutational and structural studies. The molecular mass of the purified protein is approximately 68 kDa as confirmed by mass spectrometry and it is a monomeric enzyme. Also, the enzyme assays revealed that C. glutamicum PCK has a GTP-specific activity and that its activity is maximal in the presence of both Mn2+ and Mg2+. PMID- 14550652 TI - Recombinant expression of Munc18c in a baculovirus system and interaction with syntaxin4. AB - Two protein families that are critical for vesicle transport are the Syntaxin and Munc18/Sec1 families of proteins. These two molecules form a high affinity complex and play an essential role in vesicle docking and fusion. Munc18c was expressed as an N-terminally His-tagged fusion protein from recombinant baculovirus in Sf9 insect cells. His-tagged Munc18c was purified to homogeneity using both cobalt-chelating affinity chromatography and gel filtration chromatography. With this simple two-step protocol, 3.5 mg of purified Munc18c was obtained from a 1L culture. Further, the N-terminal His-tag could be removed by thrombin cleavage while the tagged protein was bound to metal affinity resin. Recombinant Munc18c produced in this way is functional, in that it forms a stable complex with the SNARE interacting partner, syntaxin4. Thus we have developed a method for producing and purifying large amounts of functional Munc18c--both tagged and detagged--from a baculovirus expression system. We have also developed a method to purify the Munc18c:syntaxin4 complex. These methods will be employed for future functional and structural studies. PMID- 14550653 TI - Expression of a novel recombinant dual human stem cell factor in insect cells. AB - Stem cell factor (SCF) is a hematopoietic cytokine that promotes the survival, proliferation, and differentiation of hematopoietic cells. A dual human stem cell factor (dhSCF) cDNA was constructed, which consisted of a full-length human stem cell factor cDNA plus a truncated hSCF cDNA (1-145aa), linked by a peptide (GGGGSGGGGSGG) coding region. The dhSCF gene was cloned into baculovirus transfer vector pAcSecG2T under the control of polyhedrin promoter. The Sf9 cells infected with the recombinant virus expressed rdhSCF up to 6000 U/10(6) cell in flask and 8300 U/10(6) cell in spinner flask. The rdhSCF was purified by two-step chromatography. The molecular mass of rdhSCF was examined by western blotting and HPLC analysis. The specific activity of rdhSCF was up to 3.1x10(6) U/mg, about 8.7 times as high as that of monomer rhSCF from Escherichia coli. PMID- 14550654 TI - An integrated strategy for sustainable forest-energy-environment interactions in Nigeria. AB - The Nigerian forests have been subjected to unguarded exploitation over the years. Although there is overwhelming empirical evidence, which show that Nigeria's forest, may soon vanish, available statistics have shown its increasing importance in the energy sector. With increasing population come the attendant demands on the biotic environment through increased land clearing, deforestation, devegetation, decertification, with attendant soil erosion, flooding, sand dune formation, and changes in the micro-climate with consequent loss of biological productivity and associated socio-economic and socio-political problems in the country. There is therefore the need to adopt measures that will shift the attention of the Nigerian populace from the forest to satisfy their energy needs. However, such measures that will address the challenges confronting the forestry, forest-based energy systems and the environment should be consistent with the development needs, resources and priorities of the nation. Hence, for sustainable forest-energy-environment interactions, a holistic and integrated strategy that can be adopted to minimise the observed forest depletion must take cognisance of options from various land use practices, energy and forest sectors. The focus of this paper is on a strategy of options from both the energy and forest sectors. Based on the socio-economic, socio-political and environmental analyses of various options from the energy and forest sectors, the philosophy behind the mosaic approach to sustainable development has been considered in developing the proposed strategy. Policy measures to implement this strategy of options in the national development programs are also suggested. PMID- 14550655 TI - Some comparative factors regarding recycling collection systems in regions of the USA and Europe. AB - This paper analyses whether differences in selective waste collection systems used commonly in Europe and America influence people's recycling habits and their opinions about the service they are offered. We study certain characteristics of the collection systems used in two specific areas-the Principality of Asturias (Northern Spain) and El Paso county (USA)-with very different practices. It likewise considers the extent to which such differences influence the recycling habits of the public at large, as well as their perceptions of the waste collection services. In order to carry out the research and to compare the influence of the above-mentioned factors in each case, personal interviews were held in the street in the two areas under study (1066 and 1068 interviews, respectively). Asturians needed less time to dispose of waste and separate items, being one of the reasons why this group has developed the habit of selective separation to a greater degree than El Pasoans. PMID- 14550656 TI - Developing environmental quality standards for various pesticides and priority pollutants for French freshwaters. AB - The French Ministry of the Environment and Water Agencies have derived environmental quality standards following the existing French framework called SEQ-Eau, for 28 pesticides and seven priority substances listed in the Water Framework Directive. Like other existing frameworks, SEQ-Eau relies upon the use of standard toxicity test results and assessment factors. This approach made it possible to derive the desired quality standards, but due to the lack of chronic toxicity data many of the standards are only provisional. In addition, emerging issues such as endocrine disruption should be taken into account in this framework. Depending on the available data, more flexible derivation approaches, such as the statistical distribution of NOECs, are recommended for future versions of SEQ-Eau. A comparison is made using SEQ-Eau and the methodology currently being discussed in Europe to develop environmental quality standards, showing that the European approach is more conservative. Conversely, the proposed approach under the Water Framework Directive does not cover the entire range of required quality classes. PMID- 14550657 TI - Biohydrogen production from starch in wastewater under thermophilic condition. AB - Batch experiments were conducted to convert starch in wastewater into hydrogen at 55 degrees C at various wastewater pH (4.0-9.0) and starch concentrations (9.2 36.6 g/l). The maximum hydrogen yield of 92 ml/g of starch added (17% of the theoretical value) was found at wastewater pH 6.0, and the maximum specific hydrogen production rate of 365 ml/(g-VSS.d) was at wastewater pH 7.0. The methane-free biogas contained up to 60% of hydrogen. The mixed liquor was composed mostly of acetate (40.2-53.4%) and butyrate (26.0-40.9%). Phylogenetic analysis based on 16S rDNA sequences of the 72 clones developed from the sludge at pH 6.0 shows that 85.7% of the clones were closely affiliated with genus Thermoanaerobacterium in family Thermoanaerobacteriaceae; the remaining 14.3% were with an uncultured Saccharococcus sp. clone ETV-T2. PMID- 14550658 TI - Numerical modeling of diazinon transport through inter-row vegetative filter strips. AB - A numerical simulation model of pesticide runoff through vegetative filer strips (PRVFS) was developed as a tool for investigating the effects of pesticide transport mechanisms on VFS design in dormant-sprayed orchard. The PRVFS model was developed applying existing theories such as kinematic wave theory and mixing zone theory for pesticide transport in the bare soil area. For VFS area, the model performs flow routing by simple mass accounting in sequential segments and the pesticide mass balance by considering pesticide washoff and adsorption processes on the leaf, vegetative litter, root zone and soil. Model sensitivity analysis indicated that pesticide transfer from surface soil to overland flow and pesticide washoff from the VFS were important mechanisms affecting diazinon transport. The VFS cover ratio and rainfall intensity can be important design parameters for controlling diazinon runoff using inter-row VFS in orchard. The PRVFS model was validated using micro-ecosystem simulation of diazinon transport for 0, 50 and 100% VFS cover conditions. The PRVFS model is shown to be a beneficial tool for evaluating and analyzing possible best management practices for controlling offsite runoff of dormant-sprayed diazinon in orchards during the rainy season. PMID- 14550659 TI - Heterogeneous photocatalysed degradation of a herbicide derivative, isoproturon in aqueous suspension of titanium dioxide. AB - Heterogeneous photocatalysed degradation of a herbicide derivative, N-(4 isopropylphenyl)-N',N'-dimethylurea (Isoproturon, 1) was investigated in aqueous suspensions of titanium dioxide by monitoring the change in absorption intensity and depletion in Total Organic Carbon content as a function of irradiation time. The degradation kinetics was studied under different conditions such as pH, catalyst concentration, substrate concentration, different types of TiO(2) and in the presence of electron acceptors such as hydrogen peroxide (H(2)O(2)), potassium bromate (KBrO(3)) and potassium persulphate (K(2)S(2)O(8)) besides molecular oxygen. The degradation rates were found to be strongly influenced by all the above parameters. The photocatalyst Degussa P25 was found to be more efficient as compared with other photocatalysts. An attempt was made to identify the degradation product through GC-MS analysis technique. PMID- 14550660 TI - Industrial-scale demonstration of a new sorbent reactivation technology for fluidized bed combustors. AB - To minimize the disposal of highly reactive spent sorbent from a fluidized bed combustor, a new method for reactivation has been developed. The method consists of grinding the spent ash in a rotary mill, hydrating the ash with an excess of water, and mixing the wet ground ash with dry solids to absorb the excess water. The mixing process eliminates the formation of a concrete-like product that normally results as wet fluidized bed combustor ash ages. Pilot-scale combustion trials proved to be successful, and the process was scaled up using a 35MWt utility boiler at Purdue University. The test lasted for 3 days and resulted in net reduction of limestone sorbent use of 18%. The results generated in this work have been used to develop an economic evaluation for a 165MWe circulating fluidized bed (CFB) boiler, which projects significant savings due to reduction of limestone supply and ash disposal costs. The evaluation also suggests that the process is cost competitive with other processes, albeit that those processes have not been demonstrated at industrial scale. Furthermore, it also has the potential to make a small net reduction in CO(2) emissions, due to reduced limestone usage. PMID- 14550661 TI - Removal of heavy metals in rinsing wastewater from plating factory by adsorption with economical viable materials. AB - The removal of heavy metals from plating factory wastewater with economical materials was investigated by the column method. Montmorillonite, kaolin, tobermorite, magnetite, silica gel and alumina were used as the economical adsorbents to wastewater containing Cd(II), Cr(VI), Cu(II) and Pb(II). This removal method of heavy metals proved highly effective as removal efficiency tended to increase with increasing pH and decrease with increasing metal concentration. The removal percentages by adsorption onto montmorillonite, tobermorite, magnetite, and silica gel showed high values for all metals. From the results for the heat of adsorption, the adsorption process in the present study might be chemisorption. The proposed method was successfully applied to the removal of Cd(II), Cr(VI) and Cu(II) in rinsing wastewater from plating factory in Nagoya City, Aichi Prefecture, Japan. Since the economical adsorbents used can be obtained commercially because they are easily synthesized, the wastewater treatment system developed is rapid, simple and cheap for the removal of heavy metals. PMID- 14550662 TI - Fed-batch and batch operating mode analysis of a stirred anaerobic sequencing reactor with self-immobilized biomass treating low-strength wastewater. AB - This work presents an analysis of a stirred anaerobic sequencing discontinuous reactor with different substrate feeding strategies resulting in batch, fed batch/batch and fed-batch operating modes. The reactor, containing granulated biomass, was fed with approximately 2.0L of synthetic domestic wastewater with Chemical Oxygen Demand of nearly 500 mg/L per cycle and operated at 30 degrees C and 50 rpm. Three feeding strategies with a total cycle time of 6 h, including 30 min settling, were adopted: batch mode with a fill cycle of 6 min, a fed batch/batch mode with fill cycles of 60, 120 and 240 min and fed-batch mode with a fill cycle of 320 min. The system attained average non-filtered and filtered substrate removal efficiency of 78 and 84%, respectively, for all operating conditions, presenting good stability, solid retention and no granule break-up. A first order kinetic model with a residual organic matter concentration was proposed to analyze the influence of the feeding strategy on the performance during a cycle and bicarbonate alkalinity and total volatile acids concentration profiles were also quantified in order to verify the transient stability behavior. PMID- 14550663 TI - Boater preferences for beach characteristics downstream from Glen Canyon Dam, Arizona. AB - Release flow decisions are increasingly being influenced by an array of social values, including those related to river-based recreation. A substantial portion of past recreation research on downstream impacts of dams has focused on variability of instream flows. This study complements past research by assessing user preferences for beach characteristics affected by long-term impacts of flow regimes. Based upon a study of three recreational user groups (private trip leaders, commercial passengers, and river guides) of the Colorado River in Grand Canyon, preferences for beach size, presence of shade on beach, and presence of vegetation on beach are examined. Results indicate that large size beaches with shade from trees are setting characteristics with highly reliable and strong user preferences. The multinomial regression models developed for each user group indicate that 80% of all respondents would choose beach campsites 800 m(2); results were the same regardless of respondents' past boating experience, boat type (i.e. oar or motorized), or group size. In addition, size of beach was consistently reported to be a trip feature of moderate importance to respondents' river trip. Implications of this research are related to future prospects for controlled floods (i.e. spike flows) released from Glen Canyon Dam. PMID- 14550664 TI - Peripheral blood and head kidney leucocyte populations during out-of-season (0+) parr-smolt transformation and seawater transfer of Atlantic salmon (Salmo salar L.). AB - Using monoclonal antibodies (MAb) and flow cytometry, Atlantic salmon neutrophils and Ig+ cells in blood and head kidney were studied in under-yearling out-of season (0+) smolts, and 2 and 4 weeks after transfer to seawater. The parr-smolt transformation was induced using a phase advanced simulated natural photoperiod regime, and sampling of four fish was performed at regular intervals, starting on the date of the photoperiod initiation. During the freshwater period the proportion of neutrophils in the head kidney leucocytes (HKL) remained quite stable and only gradual changes in Ig+ cells were observed. In the peripheral blood leucocytes (PBL), the proportion of neutrophils markedly increased during the last month prior to seawater transfer. The most notable changes in the proportions of MAb+ leucocytes were observed in PBL after seawater transfer, with a significant increase in Ig+ cells and a significant decrease in neutrophils after two weeks in seawater. In the freshwater samples, although there were fluctuations, a decrease in the numbers of total leucocytes per millilitre blood and per gram head kidney during parr-smolt transformation was observed. The number of MAb+ cells in blood appeared to be relatively stable, while the number in head kidney tended to decrease. Following seawater transfer, the numbers of total and MAb+ leucocytes in both blood and head kidney increased markedly. The results suggest that changes in both distribution and numbers of leucocytes in peripheral blood and head kidney take place during parr-smolt transformation, and that marked changes are associated with seawater transfer. Some mechanisms possibly involved are indicated. PMID- 14550665 TI - Association of Flavobacterium psychrophilum with rainbow trout (Oncorhynchus mykiss) kidney phagocytes in vitro. AB - The capacity of virulent and non-virulent strains of Flavobacterium psychrophilum of different serotypes to associate with isolated rainbow trout (Oncorhynchus mykiss, 300-500 g) kidney phagocytes was evaluated in vitro. The results showed that F. psychrophilum was associated with the phagocytes but large differences in association were observed between the different bacterial strains examined. These differences in association with the phagocytes was not clearly related to the serotype or virulence of the bacteria, although all strains tested of the non virulent serotype FpT showed strong association with the isolated phagocytes. A competitive association assay with treatment of the phagocytes with seven different carbohydrates, suggested a role for N-acetylneuraminic acid (sialic acid) in the binding of F. psychrophilum to phagocytes. A significant dose dependent inhibition of the association was observed with sialic acid. Treatment of F. psychrophilum with sodium-metaperiodate showed that carbohydrate components play a role in the adhesion of the bacteria to the phagocytes. The results indicate that the binding of F. psychrophilum to rainbow trout kidney phagocytes can be mediated by opsonin independent cell-receptor adhesion. All tested strains seemed to be non-cytotoxic for rainbow trout kidney phagocytes in vitro suggesting that a phagocyte toxin is not necessary for the virulence of F. psychrophilum PMID- 14550667 TI - The functional characterisation of CK-1, a putative CC chemokine from rainbow trout (Oncorhynchus mykiss). AB - Recently a number of cytokine homologs have been cloned in teleost fish, including several that resemble chemokines, but to date few have been confirmed using functional assays. Chemokines are a family of cytokines that are able to induce chemotaxis in leucocytes. In this study CK-1, a rainbow trout chemokine, was functionally characterised. Recombinant CK-1 is able to attract rainbow trout peripheral blood leucocytes (PBL) in a micro-chemotaxis chamber. A greater number of PBLs migrated in response to CK-1 than to negative controls, either media alone or equivalent concentrations of beta2M, while comparable numbers migrated to the positive control, recombinant human C5a. The tissue distribution of CK-1 mRNA was also assessed by Northern blotting of RT-PCR and showed that expression is constitutive in the liver and gut, and is inducible by intraperitoneal injection of phytohemagglutinin in PBL and the head-kidney. Continuous cell lines generated from the gut and pituitary gland of the rainbow trout also express CK-1 message, whilst Southern analysis shows that CK-1 is a single copy gene. Finally, CK-1 shows the greatest amino acid similarity CCL20/LARC/Mip-3alpha as well as similar gene structure and expression pattern. PMID- 14550666 TI - Multiple acute temperature stress affects leucocyte populations and antibody responses in common carp, Cyprinus carpio L. AB - Stress is a potential factor causing increased susceptibility of fish to pathogens. In this study, stress-induced immunological changes that may contribute to a decreased immune status were investigated. A 3 h drop in ambient water temperature of 9 degrees C was used as a relative mild and acute stress model for carp. Effects of this stressor on the dynamics of leucocyte populations were determined with specific monoclonal antibodies. The relative number of circulating B-lymphocytes in the total leucocyte population decreased significantly within 4 h after the onset of single or multiple cold shocks. This decrease was reversible, as B-lymphocyte numbers were restored within 24 h. Most probably, a redistribution of B-lymphocytes contributed to this phenomenon. In head kidney, an increase was measured in the relative number of B-lymphocytes. Granulocyte numbers showed opposite reactions: the percentage of granulocytes in the total leucocyte population nearly doubled in circulation and decreased significantly in the head kidney. This demonstrates that in vivo, a mild stressor differentially alters the distribution of leucocytes. In stressed carp, the percentage of apoptotic lymphocytes in blood is significantly higher compared with the unstressed animals. B-lymphocytes as well as Ig- lymphoid cells contributed to this increased apoptosis. Labelling of blood lymphocytes with a polyclonal antiserum against the glucocorticoid receptor also showed, besides B lymphocytes, part of the Ig- lymphoid cell population to be glucocorticoid receptor positive. As the distribution of B-lymphocytes was substantially affected, the effect of temperature stress on T-lymphocyte-independent (trinitrophenyl-lipopolysaccharide) and T-lymphocyte-dependent (dinitrophenyl keyhole limpet hemocyanin) humoral antibody responses was determined. Kinetics of the primary antibody response to the T-lymphocyte-independent antigen showed lower antibody titres in stressed carp during the onset of the immune response, implying a slower development of the antibody response against the T-lymphocyte independent antigen. PMID- 14550668 TI - Streptococcus iniae expresses a cell surface non-immune trout immunoglobulin binding factor when grown in normal trout serum. AB - Three capsulated isolates of S. iniae representing serotype I and II and being arginine dihydrolase positive, negative or variable (AD+ve, AD-ve, AD+-ve) were investigated for their ability to bind rainbow trout serum immunoglobulin by the Fc region. Using a coagglutination assay with bacteria grown in Todd-Hewitt broth (THB), no evidence of non-specific Fc-binding of trout immunoglobulin (Ig) was obtained. However, when grown in normal trout serum, all isolates produced similar protein patterns in SDS-PAGE, but they were markedly different from the patterns of the bacteria grown in THB. Some bands with MW 70 kDa and over 100 kDa were very intense in the profiles of the serum-grown isolates. In Western blots, these bands of all isolates were immunostained with the conjugated goat antiserum to trout Ig, after blocking with normal goat serum, demonstrating that the bacteria had bound the trout Ig during growth in the serum. When the isolates were grown overnight in trout antiserum against Lactococcus garvieae they coagglutinated with L. garvieae cells but S. iniae isolates grown in normal trout serum did not. These data indicate that S. iniae grown in serum express surface factors which can bind trout Ig by the Fc-region. PMID- 14550669 TI - Effects of different stressor agents on gilthead seabream natural cytotoxic activity. AB - Several common situations in gilthead seabream (Sparus aurata L.) farming, such as air exposure, crowding and the use of anaesthetics, have been demonstrated to be stressful. In the present study, these conditions were simulated in the laboratory, after which head-kidney natural cytotoxic cell (NCC) activity was evaluated. For this, several specimens were air exposed for 2 min, returned to the aquarium and sampled from 0 to 4 days after exposure. NCC activity was significantly lower on the day following air-exposure compared with the control (rested fish) but not at any other time studied. Other fish were crowded (100 kg biomass m(-3), 2 h), returned to an aquarium with the same density as the control group (9 kg m(-3)) and sampled from 0 to 4 days after treatment. Head-kidney NCC activity was statistically increased compared with the control (resting) fish, 1 day after crowding. Anaesthesis for 1 h with 60 or 200 microl 2-phenoxyethanol l( 1)had no significant effect on NCC activity, while the use of 50 mg MS222 l( 1)for 1 h reduced such activity (by about 40%) compared with the control. In other experiments, fish were consecutively treated with crowding and anaesthetics. When treated with the lowest 2-phenoxyethanol concentration after crowding, the NCC activity inhibition was abolished compared with the activity in fish treated either with crowding or anaesthetic alone, while the use of the highest concentration increased such inhibition. The use of MS222 after crowding did not produce any differential effect compared with the fish treated with only one of the factors. In conclusion, NCC activity is affected differently according to the stress factor applied (hypoxia, crowding and/or anaesthetics). Differences in the effects provoked by these stressors on other seabream innate immune parameters are discussed. PMID- 14550670 TI - Immune enhancement in rainbow trout (Oncorhynchus mykiss) by potential probiotic bacteria (Lactobacillus rhamnosus). AB - The present study assessed the immune enhancement of fish by a lactic acid bacterium (LAB) Lactobacillus rhamnosus (ATCC 53103). The bacterium was administered orally at five different doses 7.9 x 10(4) (LAB4), 2.1 x 10(6) (LAB6), 2.8 x 10(8) (LAB8), 1.9 x 10(10) (LAB10) and 9.7 x 10(10) (LAB11) CFU/g feed to rainbow trout for two weeks and the feed was changed to un-supplemented diet. From the onset of feeding supplemented diets at 1, 2, 3 and 4 weeks, blood and mucus samples were taken. During the LAB feeding period L. rhamnosus persisted in the fish intestine and in the tank water in high numbers. However, L. rhamnosus disappeared from the intestine, skin mucus and tank water within one week after the change to the non-supplemented feed. In comparison to untreated control fish, respiratory burst activity of blood cells was raised significantly in the LAB4 treated group on week 2. Serum-mediated killing of Escherichia coli was increased significantly in group LAB6 on week 2. Serum immunoglobulin levels were significantly raised only in LAB8 group on week 1 and in LAB4 and LAB8 at the end of the trial. The results show that rainbow trout immune parameters were enhanced by using probiotic bacteria. PMID- 14550672 TI - Stimulation of antioxidant enzymes levels in carp (Cyprinus carpio L.) infected by Ptychobothrium sp. (Cestoda). AB - Increased antioxidant enzymatic activities were observed in carp parasitised by Ptychobothrium sp. when compared with healthy fish. This antioxidant response could contribute to neutralise the oxidative stress normally induced by parasitism. PMID- 14550671 TI - Molecular cloning and characterization of crucian carp (Carassius auratus L.) interferon regulatory factor 7. AB - Interferon (IFN) can induce an antiviral state via interferon-regulatory transcription factors (IRFs), which bind to and control genes directed by the interferon-stimulated response element (ISRE). Here we describe a fish IRF, termed CaIRF7, cloned from a subtractive cDNA library which is constructed with mRNAs obtained from crucian carp (Carassius auratus L.) blastulae embryonic (CAB) cells infected by UV-inactivated GCHV and mock-infected cells. CaIRF7 cDNA was found to be 1816 bp in length, with a 42 bp 5'UTR and a 508 bp 3'UTR. The open reading frame translates into 421 amino acids in which a DNA-binding domain (DBD) containing the repeated tryptophan motif and IRFs association domain have been identified. Like chicken GgIRF3, CaIRF7 was most similar to mammalian IRF7 with 27 to 30% identity overall and some 37% identity in their DBDs. A single transcript of 1.9 kb was detected in virally induced CAB cells by virtual Northern blotting. RT-PCR analysis revealed a wide tissue distribution of CaIRF7 constitutive expression, with detectable transcript in non-infected CAB cells and various tissues of healthy crucian carp. In addition, CaIRF7 expression was differentially increased by stimulation of the CAB cells with active GCHV, UV inactivated GCHV or CAB IFN, indicating that the activation of CaIRF7 was directly regulated by IFN. PMID- 14550673 TI - The promoter for the Interferon Regulatory Factor (IRF)-2 in the rainbow trout Oncorhynchus mykiss: cloning and reporter gene activity. PMID- 14550674 TI - Annual variation of complement, lysozyme and haemagglutinin levels in serum of the gilthead sea bream Sparus aurata. PMID- 14550675 TI - Mitogenic responses of leukocytes from the spotted wolffish (Anarhichas minor Olafsen). PMID- 14550676 TI - Alternative splicing in the nervous system: an emerging source of diversity and regulation. AB - Alternative splicing is emerging as a major mechanism of functional regulation in the human genome. Previously considered to be an unusual event, it has been detected by many genomics studies in 40%-60% of human genes. Moreover, it appears to be of central importance for neuronal genes and other genes involved in "information processing" functions. In this review, we will summarize alternative splicing's effects on mRNA transcripts, protein products, biological function, and human disease, focusing on genes of neuropsychiatric interest. We will also describe the latest experimental methods and database resources that can help neuroscientists make use of alternative splicing in their own research. PMID- 14550677 TI - Neural abnormalities during cognitive generation of affect in treatment-resistant depression. AB - BACKGROUND: Dysfunctions in brain regions known to be involved in affect and mood states are thought to be implicated in depression and may have a role in determining the type and symptoms of this illness. METHODS: Functional magnetic resonance imaging was used to elucidate neural correlates of cognitive generation of affect, using a previously published paradigm of evoking affect with picture caption pairs, in patients with unipolar, treatment-resistant depression. RESULTS: Compared with control participants, patients showed relatively decreased response in the anterior cingulate (rostral; right) with both negative and positive picture-caption pairs and in the medial frontal gyrus and hippocampus (all left) with positive picture-caption pairs. They demonstrated increased response in the inferior (right) and middle temporal gyri (left) with negative picture-caption pairs, and in the parahippocampal gyrus (right), inferior frontal gyrus (left), subgenual cingulate (right), striatum (right), and brain stem (left) with positive picture-caption pairs. CONCLUSIONS: Reduced medial/middle prefrontal and hippocampal activity may account for positive affect disturbances and temporal lobe hyperactivity for negative affect disturbances in treatment resistant depression. The results also corroborate previous observations from resting positron emission tomography studies and further elucidate the association between hypoactive rostral cingulate and nonresponsiveness to treatment in depression. PMID- 14550678 TI - Cerebral benzodiazepine receptors in depressed patients measured with [123I]iomazenil SPECT. AB - BACKGROUND: A recent magnetic resonance spectroscopy (MRS) study revealed low gamma-aminobutyric acid (GABA) levels in the occipital cortex of depressed patients. No in vivo study has been reported to measure postsynaptic GABA receptors in the patients. METHODS: Cortical benzodiazepine (BZ) binding to GABA(A) receptors was measured with [(123)I]iomazenil and single photon emission computed tomography in unmedicated patients with unipolar major depression (n = 13) and healthy subjects (n = 19). Group differences were evaluated by means of statistical parametric mapping (SPM) with partial volume correction for gray matter. Occipital GABA levels were determined by proton MRS in a subgroup (n = 6) of the patients. RESULTS: No evidence of altered BZ binding was found in patients with depression compared with healthy control subjects in the SPM analysis. Although reduction in gray matter volume was observed in the frontal cortex and amygdala of the patients, partial volume correction of the atrophy did not change the result of unaltered BZ binding. GABA levels were found lower in the occipital cortex; however, BZ binding did not show significant relationship to GABA levels. CONCLUSIONS: GABA(A) receptor binding measured in vivo with BZ radioligand binding are not altered in patients with depression. PMID- 14550679 TI - In vivo activity of bupropion at the human dopamine transporter as measured by positron emission tomography. AB - BACKGROUND: Converging lines of evidence are consistent with an inhibitory effect of the antidepressant and smoking-cessation aid bupropion on dopamine and norepinephrine reuptake, but the in vivo effects of the drug at the human dopamine transporter (DAT) have not been studied to date. This study employed positron emission tomography (PET) to assess the extent and duration of DAT receptor occupancy by bupropion and its metabolites under conditions of steady state oral dosing with bupropion sustained-release (SR) in healthy volunteers. METHODS: Six healthy male volunteers received bupropion SR 150 mg daily on days 1 through 3 and 150 mg every 12 hours on day 4 through the morning of day 11. PET investigations were performed between 1 and 7 days before initiation of bupropion SR dosing, as well as 3, 12, and 24 hours after the last dose of bupropion SR on day 11. RESULTS: Bupropion and its metabolites inhibited striatal uptake of the selective DAT-binding radioligand (11)C-betaCIT-FE in vivo. Three hours after the last dose of bupropion SR, average DAT occupancy by bupropion and its metabolites was 26%-a level that was maintained through the last PET assessment at 24 hours after dosing. CONCLUSIONS: Bupropion and its metabolites induced a low occupancy of the striatal DAT over 24 hours under conditions of steady-state oral dosing with therapeutic doses of bupropion SR. These data are consistent with the hypothesis that dopamine reuptake inhibition may be responsible in part for the therapeutic effects of the drug. PMID- 14550681 TI - Intensity-dependent regional cerebral blood flow during 1-Hz repetitive transcranial magnetic stimulation (rTMS) in healthy volunteers studied with H215O positron emission tomography: I. Effects of primary motor cortex rTMS. AB - BACKGROUND: Repetitive transcranial magnetic stimulation (rTMS) affects the excitability of the motor cortex and is thought to influence activity in other brain areas as well. We combined the administration of varying intensities of 1 Hz rTMS of the motor cortex with simultaneous positron emission tomography (PET) to delineate local and distant effects on brain activity. METHODS: Ten healthy subjects received 1-Hz rTMS to the optimal position over motor cortex (M1) for producing a twitch in the right hand at 80, 90, 100, 110, and 120% of the twitch threshold, while regional cerebral blood flow (rCBF) was measured using H(2)(15)O and PET. Repetitive transcranial magnetic stimulation (rTMS) was delivered in 75 pulse trains at each intensity every 10 min through a figure-eight coil. The regional relationship of stimulation intensity to normalized rCBF was assessed statistically. RESULTS: Intensity-dependent rCBF increases were produced under the M1 stimulation site in ipsilateral primary auditory cortex, contralateral cerebellum, and bilateral putamen, insula, and red nucleus. Intensity-dependent reductions in rCBF occurred in contralateral frontal and parietal cortices and bilateral anterior cingulate gyrus and occipital cortex. CONCLUSIONS: This study demonstrates that 1-Hz rTMS delivered to the primary motor cortex (M1) produces intensity-dependent increases in brain activity locally and has associated effects in distant sites with known connections to M1. PMID- 14550680 TI - Randomized, placebo-controlled trial of nefazodone maintenance treatment in preventing recurrence in chronic depression. AB - BACKGROUND: Maintenance treatment to prevent recurrences is recommended for chronic forms of major depressive disorder (MDD), but few studies have examined maintenance efficacy of antidepressants with chronic MDD. This randomized, placebo-controlled study of the efficacy and safety of nefazodone in preventing recurrence was conducted for patients with chronic MDD. METHODS: A total of 165 outpatients with chronic, nonpsychotic MDD, MDD plus dysthymic disorder ("double depression"), or recurrent MDD with incomplete inter-episode recovery, who achieved and maintained a clinical response during acute and continuation treatment with either nefazodone alone or nefazodone combined with psychotherapy, were randomized to 52 weeks of double-blind nefazodone (maximum dose 600 mg/day) or placebo. The occurrence of major depressive episodes during maintenance treatment was assessed with the 24-item Hamilton Rating Scale for Depression, a DSM-IV MDD checklist, and a blinded review of symptom exacerbations by a consensus committee of research clinicians. RESULTS: Application of a competing risk model that estimated the conditional probability of recurrence among those patients remaining on active therapy revealed a significant (p =.043) difference between nefazodone (n = 76) and placebo (n = 74) when the latter part of the 1 year maintenance period was emphasized. At the end of 1 year, the conditional probability of recurrence was 30.3% for nefazodone-treated patients, compared with 47.5% for placebo-treated patients. Prior concomitant psychotherapy during acute/continuation treatment, although enhancing the initial response, was not associated with lower recurrence rates. Discontinuations due to adverse events were relatively low for both nefazodone (5.3%) and placebo (4.8%). Somnolence was significantly greater among the patients taking active medication (15.4%), compared with placebo (4.6%). CONCLUSIONS: Nefazodone is well-tolerated and is an effective maintenance therapy for chronic forms of MDD. PMID- 14550682 TI - Intensity-dependent regional cerebral blood flow during 1-Hz repetitive transcranial magnetic stimulation (rTMS) in healthy volunteers studied with H215O positron emission tomography: II. Effects of prefrontal cortex rTMS. AB - BACKGROUND: The changes in brain activity produced by repetitive transcranial magnetic stimulation (rTMS) of the prefrontal cortex (PFC) remain unclear. We examined intensity-related changes in brain activity with positron emission tomography (PET) in normal volunteers during rTMS delivered to the left PFC. METHODS: In 10 healthy volunteers, we delivered 1-Hz rTMS at randomized intensities over left PFC with a figure-eight coil. Intensities were 80, 90, 100, 110, and 120% of the right-hand muscle twitch threshold. Regional cerebral blood flow (rCBF) scans were acquired with H(2)(15)O PET during rTMS at each intensity. RESULTS: Repetitive transcranial magnetic stimulation intensity was inversely correlated with rCBF in the stimulated and contralateral PFC, ipsilateral medial temporal lobe, both parahippocampi, and posterior middle temporal gyri. Positive correlations of rCBF with intensity occurred in ipsilateral anterior cingulate, cerebellum, contralateral insula, primary auditory cortex, and somatosensory face area. CONCLUSIONS: The intensity-related inverse relationship between 1-Hz rTMS and prefrontal activity appears opposite to that seen with rTMS over the motor cortex in a companion study. Intensity-dependent increases in rCBF were seen in a number of distant cortical and subcortical areas with PFC rTMS, suggesting activation of left anterior cingulate, claustrum, and cerebellum. The regional differences in direction of rTMS effects and the greater activation of distant structures at higher intensities suggest the potential importance of higher intensity prefrontal rTMS for therapeutic applications in neuropsychiatric patients. PMID- 14550683 TI - S-adenosyl-L-methionine: effects on brain bioenergetic status and transverse relaxation time in healthy subjects. AB - BACKGROUND: S-adenosyl-L-methionine is an effective treatment for clinical depression, although the mechanism underlying this effect is unclear. Presently, in vivo phosphorus magnetic resonance spectroscopy (31P MRS) and brain transverse relaxometry were employed to test if S-adenosyl-L-methionine supplementation alters brain bioenergetics and/or transverse relaxation time (T2RT) in a nondepressed cohort. If these magnetic resonance techniques are sensitive to S adenosyl-L-methionine induced alterations in neurochemical processes, these methods may be used in cases of clinical depression to elucidate the mechanism underlying the antidepressant effect of S-adenosyl-L-methionine. METHODS: Twelve subjects self-administered 1600 mg of oral S-adenosyl-L-methionine daily. Phosphorus spectra and transverse relaxation time were acquired at baseline and after treatment using a 1.5 Tesla scanner. RESULTS: Phosphocreatine levels were significantly higher after treatment, whereas beta nucleoside triphosphate levels, predominantly adenosine triphosphate in brain, were significantly lower after treatment. A surprising gender difference in T2RT emerged after supplementation, with women exhibiting significantly lower T2RT than men. CONCLUSIONS: Alterations in phosphocreatine and beta nucleoside triphosphate are consistent with the report that S-adenosyl-L-methionine is involved in the production of creatine, which in turn is phosphorylated to phosphocreatine using adenosine triphosphate. These findings suggest that S-adenosyl-L-methionine alters parameters associated with cerebral bioenergetic status and that some effects of S-adenosyl-L-methionine (T2RT) occur in a gender-specific manner. PMID- 14550684 TI - Differential enhancement of antidepressant penetration into the brain in mice with abcb1ab (mdr1ab) P-glycoprotein gene disruption. AB - BACKGROUND: Mice with a genetic disruption (knockout) of the multiple drug resistance (abcb1ab) gene were used to examine the effect of the absence of the drug-transporting P-glycoprotein (P-gp) at the blood-brain barrier on the uptake of the antidepressants venlafaxine, paroxetine, mirtazapine, and doxepin and its metabolites into the brain. METHODS: One hour after subcutaneous injection of venlafaxine, paroxetine, mirtazapine, or doxepin, knockout and wildtype mice were sacrificed, and the drug concentrations in brain, spleen, kidney, liver, and plasma were measured. RESULTS: The cerebrum concentrations of doxepin, venlafaxine, and paroxetine were higher in knockout mice, demonstrating that these substances are substrates of P-gp and that abcb1ab activity at the level of the blood-brain barrier reduces the penetration of these substances into the brain. In contrast, brain distribution of mirtazapine was indistinguishable between the groups. CONCLUSIONS: The differences reported here in brain penetration of antidepressant drugs that depend on the presence of the abcb1ab gene may offer an explanation for poor or nonresponse to antidepressant treatment. Furthermore, they may be able to explain in part the discrepancies between plasma levels of an antidepressant and its clinical effects and side effects. PMID- 14550685 TI - Lithium alters measures of auditory gating in two strains of mice. AB - BACKGROUND: There are similarities between schizophrenia and bipolar disorder, especially during the psychotic phase. Auditory gating deficits are common in both schizophrenia (does not remit postpsychotic event) and bipolar disorder (only during the manic phase). Lithium has been used to treat psychosis acutely in both bipolar disorder and schizophrenia. An animal model was used to assess the effects of lithium treatment on normal and deficient auditory gating. METHODS: Mice of the DBA/2 (deficient gating) and C3H (normal gating) strains were treated for 6 weeks with either standard rodent chow or rodent chow supplemented with 2.55g/kg lithium carbonate. After 6 weeks of treatment, auditory evoked potentials were recorded under anesthesia. Differences between the groups and treatments were determined using analysis of variance. RESULTS: The normally impaired DBA/2 mice showed improved auditory gating following lithium treatment, while the C3H mice, the benchmark "normal" mouse strain, were impaired after lithium treatment. CONCLUSIONS: C3H mice treated with lithium had significantly impaired auditory gating as a result of treatment. This may be due to norepinephrine facilitation, through a blockade of presynaptic alpha(2) autoreceptors. DBA/2 mice had improved gating as a result of treatment with lithium, likely due to improved functioning of the gamma-aminobutyric acid system. PMID- 14550686 TI - Safety of injectable opioid maintenance treatment for heroin dependence. AB - BACKGROUND: There is a growing debate about injectable opioid treatment programs in many Western countries. This is the first placebo-controlled study of the safety of injectable opioids in a controlled treatment setting. METHODS: Twenty five opioid-dependent patients on intravenous (IV) heroin or IV methadone maintenance treatment were randomly assigned to either their individual prescribed IV maintenance dose or placebo. Acute drug effects were recorded, focusing on electrocardiography, respiratory movements, arterial blood oxygen saturation, and electroencephalography (EEG). RESULTS: After heroin injection, marked respiratory depression progressing to a Cheyne-Stokes pattern occurred. Peripheral arterial blood oxygenation decreased to 78.9 +/- 8.7% (mean +/- SD) ranging from 52%-90%. During hypoxia, 7 of the 16 subjects experienced intermittent and somewhat severe bradycardia. Five subjects exhibited paroxysmal EEG patterns. After methadone injection, respiratory depression was less pronounced than after heroin injection. No relevant bradycardia was noted. CONCLUSIONS: Opioid doses commonly prescribed in IV opioid treatment induce marked respiratory and circulatory depression, as well as occasionally irregular paroxysmal EEG activity. Further studies are needed to optimize the clinical practice of IV opioid treatment to prevent serious complications. Moreover, the extent of the observed effects raises questions about the appropriateness of IV opioid treatment in the present form. PMID- 14550688 TI - Philanthropic gifts for health: the stage is set. PMID- 14550687 TI - Neuronavigated repetitive transcranial magnetic stimulation in patients with tinnitus: a short case series. AB - BACKGROUND: Clinical as well as neurophysiological and neuroimaging data suggest that chronic tinnitus resembles neuropsychiatric syndromes characterized by focal brain activation. Low-frequency repetitive transcranial magnetic stimulation (rTMS) has been proposed as an efficient method in treating brain hyperexcitability disorders by reducing cortical excitability. METHODS: In three patients suffering from chronic tinnitus, the effect of magnetic resonance imaging and positron emission tomography guided neuronavigated 1 Hz rTMS on auditory cortex activity was evaluated, using a sham controlled double-blind crossover design. RESULTS: Two of three patients revealed clearly increased metabolic activity in circumscript areas of the primary auditory cortex (PAC), allowing a selective stimulation of these cortical areas with low-frequency rTMS. Considerable improvement in tinnitus was achieved in these patients. CONCLUSIONS: Neuronavigated rTMS of increased PAC activity may help to better understand the neuronal basis of chronic tinnitus and might offer a new option for treating auditory phantom perceptions like chronic tinnitus. PMID- 14550689 TI - Drug-delivering coronary artery stents: bare metal threatened by extinction? PMID- 14550690 TI - Birthweight and risk for cerebral palsy. PMID- 14550691 TI - Paediatric cholestasis: is villin the villain? PMID- 14550692 TI - Urban health: a new discipline. PMID- 14550693 TI - Putting Jenner back in his place. PMID- 14550694 TI - Sirolimus-eluting stents for treatment of patients with long atherosclerotic lesions in small coronary arteries: double-blind, randomised controlled trial (E SIRIUS). AB - BACKGROUND: Sirolimus-eluting stents have been developed to prevent restenosis in the treatment of coronary artery disease. We investigated the risk of restenosis with use of sirolimus-eluting stents compared with bare-metal stents to assess possible differences. METHODS: We enrolled 352 patients in whom one coronary artery required treatment, with diameter 2.5-3.0 mm and lesion length 15-32 mm. We randomly assigned patients sirolimus-eluting stents (n=175) or bare-metal stents (control, n=177). At 8 months we assessed differences in minimum lumen diameter and binary restenosis within the lesion (restenosis of > or =50% diameter, including 5 mm vessel segments proximal and distal to stented segment). Patients were also followed up for 9 months for major adverse cardiac events. Analysis was by intention to treat. FINDINGS: Stent implantation was successful in 100% of sirolimus-stent patients and 99.4% of controls. The mean diameter of treated coronary arteries was 2.55 mm (SD 0.37) and mean lesion length was 15.0 mm (6.0). Multiple stents were implanted in 170 (48%) patients. At 8 months, minimum lumen diameter was significantly higher with sirolimus-eluting stents than with control stents (2.22 vs 1.33 mm, p<0.0001). The rate of binary restenosis was significantly reduced with sirolimus-eluting stents compared with control stents (5.9 vs 42.3%, p=0.0001). Significantly fewer patients with sirolimus-eluting stents had major adverse cardiac events at 9 months than did controls (8.0 vs 22.6%, p=0.0002), due mainly to a lower need for target-lesion revascularisations (4.0 vs 20.9%, p<0.0001). INTERPRETATION: Sirolimus-eluting stents are better than bare-metal stents for treatment of single long atherosclerotic lesions in a coronary vessel smaller than 3 mm in diameter. PMID- 14550696 TI - Physical examination in the care of medical inpatients: an observational study. AB - BACKGROUND: Little is known about the clinical importance of skilled physical examination in the care of patients in hospital. METHODS: Hospital records of a systematic consecutive sample of patients admitted to a general medical inpatient service were reviewed retrospectively to determine whether physical findings by the attending physician led to important changes in clinical management. Patients with pivotal physical findings were defined by an outcomes adjudication panel as those whose diagnosis and treatment in hospital changed substantially as a result of the attending physician's physical examination. Pivotal findings were classed as validated if the resulting treatment change involved the active collaboration of a consulting specialist. Findings were classed as discoverable if subsequent diagnostic testing (other than physical examination) would probably have led to the correct diagnosis. Class 1 findings were those deemed validated but not discoverable. FINDINGS: Among 100 patients, 26 had pivotal physical findings (26%; 95% CI 18-36). 15 of these (58%; 95% CI 37-77) were validated (13 with urgent surgical or other invasive procedures) and 14 were discoverable (54%; 95% CI 33-73). Seven patients had class 1 findings (7%; 95% CI 3-14). INTERPRETATION: Physical examination can have a substantial effect on the care of medical inpatients. If replicated in other settings, these findings might have important implications for medical educators and quality improvement initiatives. PMID- 14550697 TI - Gallstone ileus. PMID- 14550698 TI - Cerebral palsy and intrauterine growth in single births: European collaborative study. AB - BACKGROUND: Cerebral palsy seems to be more common in term babies whose birthweight is low for their gestational age at delivery, but past analyses have been hampered by small datasets and Z-score calculation methods. METHODS: We compared data from ten European registers for 4503 singleton children with cerebral palsy born between 1976 and 1990 with the number of births in each study population. Weight and gestation of these children were compared with reference standards for the normal spread of gestation and weight-for-gestational age at birth. FINDINGS: Babies of 32-42 weeks' gestation with a birthweight for gestational age below the 10th percentile (using fetal growth standards) were 4-6 times more likely to have cerebral palsy than were children in a reference band between the 25th and 75th percentiles. In children with a weight above the 97th percentile, the increased risk was smaller (from 1.6 to 3.1), but still significant. Those with a birthweight about 1 SD above average always had the lowest risk of cerebral palsy. A similar pattern was seen in those with unilateral or bilateral spasticity, as in those with a dyskinetic or ataxic disability. In babies of less than 32 weeks' gestation, the relation between weight and risk was less clear. INTERPRETATION: The risk of cerebral palsy, like the risk of perinatal death, is lowest in babies who are of above average weight for-gestation at birth, but risk rises when weight is well above normal as well as when it is well below normal. Whether deviant growth is the cause or a consequence of the disability remains to be determined. PMID- 14550700 TI - A gait disorder with an unexpected twist. PMID- 14550699 TI - Abnormalities in villin gene expression and canalicular microvillus structure in progressive cholestatic liver disease of childhood. AB - BACKGROUND: The molecular basis of clinical cholestasis is a subject of intense investigation. Villin is an actin binding, bundling, and severing protein needed for maintenance of structural integrity of canalicular microvilli, in which membrane transporters required for bile secretion are located. We aimed to investigate the role of canalicular cytoskeletal proteins in three genetically unrelated children with a biliary atresia-like clinical disorder, each of whom developed liver failure requiring liver transplantation. METHODS: Explanted livers from the three patients were examined by standard pathological methods followed by transmission and cryoimmunoelectron microscopy. With archival tissue samples, a panel of cytoskeletal proteins was investigated by immunohistochemistry and western blotting, with purified canalicular membrane preparations. Villin mRNA analyses were undertaken on liver homogenates, with primers from coding regions of the human villin gene. Classic biliary atresia, other types of cholestasis, and normal livers served as controls. FINDINGS: In patients, pronounced ultrastructural deformities of canaliculi and especially of their microvilli were noted, which correlated with absence of villin protein by immunostaining of liver tissue sections and by western blot analysis. Additionally, villin mRNA was strikingly reduced or absent. These results differed greatly from those in controls. INTERPRETATION: These results suggest that the disorder described mimics biliary atresia, but structural and molecular pathological findings differ. We propose that a functional abnormality in villin gene expression is key to the mechanism of cholestasis in patients with progressive cholestasis and hepatic failure. PMID- 14550701 TI - Change in rates of cerebral atrophy over time in early-onset Alzheimer's disease: longitudinal MRI study. AB - The extent to which cerebral atrophy in Alzheimer's disease changes with time is unknown. We used multiple MRI scans to measure progression of cerebral atrophy in 12 patients with Alzheimer's disease who were followed up from a presymptomatic stage through to moderately severe dementia. Analysis with hierarchical regression models with quadratic terms in time provided evidence of increasing yearly percentage losses in brain volume. At the time when patients were judged to have mild dementia (mini-mental state examination score MMSE=23), mean yearly loss of brain volume was 2.8% (95% CI 2.3-3.3), which rose by 0.32% per year (0.15-0.50). Our findings reinforce the need for early diagnosis and therapeutic intervention in Alzheimer's disease. PMID- 14550702 TI - Vibrating insoles and balance control in elderly people. AB - Somatosensory function declines with age, and such changes have been associated with diminished motor performance. Input noise can enhance sensory and motor function. We asked young and elderly participants to stand quietly on vibrating gel-based insoles, and calculated sway parameters and random-walk variables. In our 27 participants, application of noise resulted in a reduction in seven of eight sway parameters in young participants and all of the sway variables in elderly participants. Elderly participants showed greater improvement than young people in two variables, mediolateral range (p=0.008), and critical mean square displacement (p=0.012). Noise-based devices, such as randomly vibrating insoles, could ameliorate age-related impairments in balance control. PMID- 14550703 TI - Autism. AB - Autism is a disorder characterised by severe difficulties in social interaction and communication, and with unusual behaviours. Once thought of as rare, autism is now recognised as being common. The role of CNS factors in pathogenesis is suggested by high rates of seizure disorder; research has highlighted the role of several specific brain regions in syndrome pathogenesis. Autism is a strongly genetic disorder and probably arises because of multiple genes; recurrence rates in families with one child are high. Early intervention with various techniques is helpful in many cases. Some pharmacological agents may help with certain problematic behaviours but do not address the underlying cause of the disorder. PMID- 14550704 TI - Echinocandin antifungal drugs. AB - The echinocandins are large lipopeptide molecules that are inhibitors of beta (1,3)-glucan synthesis, an action that damages fungal cell walls. In vitro and in vivo, the echinocandins are rapidly fungicidal against most Candida spp and fungistatic against Aspergillus spp. They are not active at clinically relevant concentrations against Zygomycetes, Cryptococcus neoformans, or Fusarium spp. No drug target is present in mammalian cells. The first of the class to be licensed was caspofungin, for refractory invasive aspergillosis (about 40% response rate) and the second was micafungin. Adverse events are generally mild, including (for caspofungin) local phlebitis, fever, abnormal liver function tests, and mild haemolysis. Poor absorption after oral administration limits use to the intravenous route. Dosing is once daily and drug interactions are few. The echinocandins are widely distributed in the body, and are metabolised by the liver. Results of studies of caspofungin in candidaemia and invasive candidiasis suggest equivalent efficacy to amphotericin B, with substantially fewer toxic effects. Absence of antagonism in combination with other antifungal drugs suggests that combination antifungal therapy could become a general feature of the echinocandins, particularly for invasive aspergillosis. PMID- 14550705 TI - Willingness to pay for AIDS treatment: myths and realities. PMID- 14550706 TI - Radiotherapy and tamoxifen in women with completely excised ductal carcinoma in situ. PMID- 14550707 TI - Radiotherapy and tamoxifen in women with completely excised ductal carcinoma in situ. PMID- 14550708 TI - Radiotherapy and tamoxifen in women with completely excised ductal carcinoma in situ. PMID- 14550711 TI - Radiotherapy and tamoxifen in women with completely excised ductal carcinoma in situ. PMID- 14550709 TI - Radiotherapy and tamoxifen in women with completely excised ductal carcinoma in situ. PMID- 14550712 TI - PAHO fighting AIDS. PMID- 14550714 TI - Treatment of SARS with human interferons. PMID- 14550713 TI - Bias in chlamydia prevalence surveys. PMID- 14550716 TI - Power shutdown and biological standards. PMID- 14550717 TI - Politics and science. PMID- 14550718 TI - Treatment of SARS with human interferons. PMID- 14550719 TI - Human papillomavirus in a 16th century mummy. PMID- 14550724 TI - Tony Avery interview. PMID- 14550721 TI - John Walsh. PMID- 14550726 TI - Media mania, megalomania and misleading research: the need for caution in scientific publication. PMID- 14550727 TI - Better therapies for everyday pain: exciting advances in pain management. PMID- 14550728 TI - Neurodegenerations in domestic animals: a new way forward. PMID- 14550729 TI - Assessment of motor pathways: how to breach the black box. PMID- 14550730 TI - PCV-2 infection in swine; more than just postweaning multisystemic wasting syndrome. PMID- 14550731 TI - Influence of acetylsalicylic acid and ketoprofen on canine thyroid function tests. AB - Many factors including drugs can influence thyroid function in humans, rats and dogs. Studies in humans report significant effects of non-steroidal anti inflammatory agents (NSAIDs) on thyroid function tests, which can lead to misinterpretation of the results and inappropriate therapeutic decisions. As NSAIDs are used more and more frequently in dogs, it is important to know to what extent they can influence results. Eighteen spayed female beagle dogs were randomly assigned to three treatment sequences in a 3 x 3 crossover study design with treatments consisting of acetylsalicylic acid (ASA) (25 mg/kg BW q 12 h), ketoprofen (Keto) (1 mg/kg BW q 24 h) or placebo administered for a 1-week period with a 3-week washout period between treatment periods. Blood samples for determination of total thyroxine (TT4), free thyroxine (FT4), total triiodothyronine (TT3), thyrotropin (TSH), reverse triiodothyronine (rT3), Keto and ASA concentrations were taken during each treatment period on days 0, 1, 3 and 7. During the washout period samples were taken weekly. A significant decrease in TT4 was observed as soon as 24 h after ASA administration, whereas the decrease in TT3 was less pronounced and differed significantly from the placebo only after 1 week of administration. No significant effects were found for free T4 and TSH with ASA administration. No significant effects on thyroid results were found following Keto administration. The results indicate that TT4 can be markedly decreased by ASA therapy and until the results of further studies are available, thyroid function test results should be interpreted cautiously in dogs on NSAIDs therapy. PMID- 14550733 TI - Critical differences of acute phase proteins in canine serum samples. AB - The critical difference values for acute phase proteins in canine serum samples were established on a week-to-week basis. Blood samples from 11 apparently clinically healthy dogs were collected once weekly for five consecutive weeks. For each protein the total variance of analytical results was divided into intraindividual variance (S(Intra)(2)), interindividual variance (S(Inter)(2)), and analytical variance (S(Analytical)(2)). The critical difference (d(K)) was then calculated as d(k)=22(S(Intra)(2)+S(Analytical)(2)). The critical difference values were 1.95 g/L for haptoglobin, 4.85 mg/L for C-reactive protein, and 0.016 DeltaAbs/min for ceruloplasmin. When used in conjunction with the corresponding reference interval, critical difference values can be an aid in correctly interpreting acute phase protein results, by determining whether observed differences between two consecutive measurements in individual animals are due to natural variation or due to disease therapy or experimental procedures. PMID- 14550734 TI - Abnormal synaptic protein expression in two Arabian horses with equine degenerative myeloencephalopathy. AB - Numerous swollen neurons and multiple dystrophic axons were observed in the gracillis and cuneatus nuclei of two male Arabian horses, aged six and 12 months of age, with equine degenerative myeloencephalopathy. Swollen neurons and dystrophic axons showed synaptophysin, synaptosomal-associated protein of 25 kDa, syntaxin-1 and alpha-synuclein immunoreactivity. Moreover, dystrophic axons were strongly immunopositive against the ubiquitin protein and against the anti phosphorylated 200 kDa neurofilament protein. Abnormal expression of integral synaptic vesicle, synaptic vesicle-associated presynaptic plasma membrane and cytosolic proteins, which participate in the trafficking, docking and fusion of the synaptic vesicle to the plasma membrane, suggest that severe disruption of axonal transport plays a crucial role in the pathogenesis of dystrophic axons in equine degenerative myeloencephalopathy (EDM). PMID- 14550735 TI - Standardization of transcranial magnetic stimulation in the horse. AB - The influence of coil position on the peak-to-peak amplitude and onset latency of transcranial magnetic motor evoked potentials (MMEPs) in the extensor carpi radialis and cranial tibial muscles of horses was evaluated. Seven different stimulating coil positions were obtained by constructing a frame on the forehead. Two stimulation intensities (80% and 100% of maximal stimulator output) and two different coil currents (clockwise and counter-clockwise) were tested. For both recording sites MMEPs with the shortest onset latency and the largest peak-to peak amplitude were detected when the coil was placed in the median of the forehead. There was no significant difference between left and right side recordings. The direction of the current flow in the coil had no influence on the onset latency of the MMEPs. PMID- 14550736 TI - Association of porcine circovirus 2 with porcine respiratory disease complex. AB - A retrospective study was performed on natural cases of porcine respiratory disease complex (PRDC) to determine the association and prevalence of PRDC with porcine circovirus 2 (PCV2) and other co-existing pathogens in Korea. Histologically, alveolar septa were markedly thickened by infiltrates of mononuclear cells. Moderate to marked multifocal peribronchial and peribronchiolar fibrosis were present and often extended into the airway lamina propria. Among the 105 pigs with PRDC, 85 were positive for PCV2, 66 were positive for porcine reproductive and respiratory syndrome virus (PRRSV), 60 were positive for porcine parvovirus (PPV), and 14 were positive for swine influenza virus (SIV). There were 80 co-infections and 25 single infections. A co-infection of PCV2 with another additional bacterial pathogen is frequently diagnosed in PRDC. The combination of PCV2 and Pasteurella multocida (38 cases) was most prevalent followed by PCV2 and Mycoplasma hyopneumoniae (33 cases). The consistent presence of PCV2, but lower prevalence of other viral and bacterial pathogens in all pigs examined with PRDC, has led us to speculate that PCV2 plays an important role in PRDC. PMID- 14550737 TI - Echo-assisted percutaneous renal biopsy in dogs. A retrospective study of 229 cases. AB - Renal biopsies were carried out on 229 dogs using an echo-assisted, semi automatic method. Complications arising after the biopsy were also evaluated. The biopsies were examined for the accuracy of diagnosis and the presence of histologically demonstrable primary renal pathologies. A retrospective study of the observed lesions and parenchymal echogenicity showed a correlation between these factors involving the tubulo-interstitial compartment. Histopathological renal lesions and their correlation with laboratory methods are the only parameters that allow a correct diagnostic, prognostic, and therapeutic work-up in cases of clinical kidney disease in canines. PMID- 14550739 TI - Influence of food composition on splanchnic blood flow during digestion in unsedated normal dogs: a Doppler study. AB - The effect of dietary components in gastrointestinal blood flow was investigated non-invasively in dogs. Doppler ultrasound (US) waveform analysis of the celiac artery (CA) and the cranial mesenteric artery (CMA) of eight healthy dogs was performed both in the fasting state and at 20, 40, 60 and 90 min after feeding maintenance, high protein, high carbohydrate and high fat diets. Resistive index (RI), pulsatility index (PI) and the percentage differences between measurements were calculated. In fasted dogs, the CA showed significantly (p<0.05) lower resistance than the CMA. With all diets, post-prandial RI and PI values decreased significantly in both vessels (CA: RI=-12-13%; PI=-25-29%; CMA: RI=-13-15%; PI= 27-29% [p<0.05]), and in all cases significantly earlier in the CA than in the CMA (p<0.05). With all diets except high fat the maximum decrease in RI and PI was reached in the CA at 40 min and in the CMA at 60 min. High fat diets were unique in that the maximum decrease in RI and PI in the CMA occurred earlier (at 20 min) and values were slower to return to normal (p<0.05). We conclude that Doppler US can be used in dogs to quantify significant differences in the response of splanchnic blood flow to different dietary components. PMID- 14550740 TI - The effects of isoflurane anaesthesia on some Doppler-derived cardiac parameters in the common buzzard (Buteo buteo). AB - In order to gain an initial overview of the influence of anaesthesia on the results of Doppler-derived blood flow measurements in raptors, the heart rate as well as three different sample volumes of pulsed-wave spectral Doppler-derived flow velocity (diastolic flow across the left and right atrioventricular valve, systolic flow across the aortic valve) were determined in 10 common buzzards (Buteo buteo). Measurements were taken once in conscious and once in anaesthetized birds. Anaesthesia was shown to produce significant changes in cardiac parameters recorded in the same birds whilst conscious. When comparing conscious birds with each other (with one exception for right sided ventricular inflow velocity) no correlation between the heart frequency and measured blood flow velocities was evident. This was also the case under anaesthesia. However, significant differences in these parameters were evident when comparing the results obtained before and under anaesthesia. The results suggest that the influence of anaesthesia in raptors is more than a simple reduction of heart rate and that there is also reduction in blood flow velocity. PMID- 14550741 TI - The disposition of diclofenac in camels after intravenous administration. AB - The pharmacokinetics of diclofenac was studied in camels (Camelus dromedarus) (n=6) following intravenous (i.v.) administration of a dose of 2.5 mg kg(-1) body weight. The metabolism and urinary detection time were also studied. The results obtained (median and range) were as follows: the terminal elimination half-life (t(1/2beta)) was 2.35 (1.90-2.73)h, total body clearance (Cl(T)) was 0.17 (0.16 0.21)lh kg(-1). The volume of distribution at steady state (V(SS)) was 0.31 (0.21 0.39)l(-1)kg(-1), the volume of the central compartment of the two compartment pharmacokinetic model (V(C)) was 0.15 (0.11-0.17)l kg(-1). Five metabolites of diclofenac were tentatively identified in urine and were excreted mainly in conjugate form. The main metabolite was identified as hydroxy diclofenac. Both diclofenac and hydroxy diclofenac, appear to be the main elimination route for diclofenac when administered i.v. in camels. Diclofenac could be identified up to 4 days following i.v. administration in camels using a sensitive gas chromatography/mass spectrometry (GC/MS) method. PMID- 14550744 TI - Neonatal exposure to polybrominated diphenyl ether (PBDE 153) disrupts spontaneous behaviour, impairs learning and memory, and decreases hippocampal cholinergic receptors in adult mice. AB - Neonatal exposure to polybrominated diphenyl ether (PBDE 153) disrupts spontaneous behaviour, impairs learning and memory, and decreases hippocampal cholinergic receptors in adult mice. Flame retardants are used to suppress or inhibit combustion processes in an effort to reduce the risk of fire. One class of flame retardants, polybrominated diphenyl ethers (PBDEs), are present and increasing in the environment and in human milk. The present study shows that neonatal exposure to 2,2',4,4',5,5'-hexaBDE (PBDE 153), a PBDE persistent both in environment and in human milk, can induce developmental neurotoxic effects, such as changes in spontaneous behaviour (hyperactivity), impairments in learning and memory, and reduced amounts of nicotinic receptors, effects that get worse with age. Neonatal NMRI male mice were orally exposed on day 10 to 0.45, 0.9, or 9.0 mg of PBDE 153/kg of body weight. Spontaneous behaviour (locomotion, rearing, and total activity) was observed in 2-, 4-, and 6-month-old mice, Morris water maze at an age of 6 months. The behaviour tests showed that the effects were dose response and time-response related. Animals showing defects in learning and memory also showed significantly reduced amounts of nicotinic receptors in hippocampus, using alpha-bungarotoxin binding assay. The observed developmental neurotoxic effects seen for PBDE 153 are similar to those seen for PBDE 99 and for certain PCBs. Furthermore, PBDEs appear to as potent as the PCBs. PMID- 14550745 TI - Pulmonary heat shock protein expression after exposure to a metabolically activated Clara cell toxicant: relationship to protein adduct formation. AB - Heat shock proteins/stress proteins (Hsps) participate in regulation of protein synthesis and degradation and serve as general cytoprotectants, yet their role in lethal Clara cell injury is not clear. To define the pattern of Hsp expression in acute lethal Clara cell injury, mice were treated with the Clara cell-specific toxicant naphthalene (NA), and patterns of expression compared to electrophilic protein adduction and previously established organellar degradation and glutathione (GSH) depletion. In sites of lethal injury (distal bronchiole), prior to organellar degradation (1 h post-NA), protein adduction is detectable and ubiquitin, Hsp 25, Hsp 72, and heme-oxygenase 1 (HO-1) are increased. Maximal Hsp expression, protein adduction, and GSH depletion occur simultaneous (by 2-3 h) with early organelle disruption. Hsp expression is higher later (6-24 h), only in exfoliating cells. In airway sites (proximal bronchiole) with nonlethal Clara cell injury elevation of Hsp 25, 72, and HO-1 expression follows significant GSH depletion (greater than 50% 2 h post-NA). This data build upon our previous studies and we conclude that (1) in lethal (terminal bronchiole) and nonlethal (proximal bronchiole) Clara cell injury, Hsp induction is associated with the loss of GSH and increased protein adduction, and (2) in these same sites, organelle disruption is not a prerequisite for Hsp induction. PMID- 14550746 TI - Exaggerated hepatotoxicity of acetaminophen in mice lacking tumor necrosis factor receptor-1. Potential role of inflammatory mediators. AB - Transgenic mice with a targeted disruption of the tumor necrosis factor receptor 1 (TNFR1) gene were used to analyze the role of TNF-alpha in pro- and anti inflammatory mediator production and liver injury induced by acetaminophen. Treatment of wild-type mice with acetaminophen (300 mg/kg) resulted in centrilobular hepatic necrosis. This was correlated with expression of inducible nitric oxide synthase (NOS II) and nitrotyrosine staining of the liver. Expression of macrophage chemotactic protein-1 (MCP-1), KC/gro, interleukin-1beta (IL-1beta), matrix metalloproteinase-9 (MMP-9), and connective tissue growth factor (CTGF), inflammatory mediators known to participate in tissue repair, as well as the anti-inflammatory cytokine, interleukin-10 (IL-10), also increased in the liver following acetaminophen administration. TNFR1(-/-) mice were found to be significantly more sensitive to the hepatotoxic effects of acetaminophen than wild-type mice. This was correlated with more rapid and prolonged induction of NOS II in the liver and changes in the pattern of nitrotyrosine staining. Acetaminophen-induced expression of MCP-1, IL-1beta, CTGF, and MMP-9 mRNA was also delayed or reduced in TNFR1(-/-) mice relative to wild-type mice. In contrast, increases in IL-10 were more rapid and more pronounced. These data demonstrate that signaling through TNFR1 is important in inflammatory mediator production and toxicity induced by acetaminophen. PMID- 14550747 TI - Alteration of keratinocyte differentiation and senescence by the tumor promoter dioxin. AB - Exposure to the environmental contaminant dioxin, elicits a variety of responses, which includes tumor promotion, embryotoxicity/teratogenesis, and carcinogenesis in both animals and humans. Many of the effects of dioxin are mediated by the aryl hydrocarbon receptor (AHR), a ligand-activated bHLH (basic helix-loop helix)/PAS transcription factor. We initiated this study to determine whether dioxin's tumor-promoting activities may lie in its ability to alter proliferation, differentiation, and/or senescence using normal human epidermal keratinocytes (HEKs). Here, we report that dioxin appears to accelerate differentiation as measured by flow cytometry and by increased expression of the differentiation markers involucrin and filaggrin. In addition, dioxin appears to increase proliferation as indicated by an increase in NADH/NADPH production and changes in cell cycle. Finally, dioxin decreases SA (senescence associated) beta galactosidase staining, an indicator of senescence, in the differentiating keratinocytes. These changes were accompanied by decreases in the expression levels of key cell cycle regulatory proteins p53, p16INK4a, and p14ARF. Our findings support the idea that dioxin may exert its tumor-promoting actions, in part, by downregulating the expression levels of key tumor suppressor proteins, which may impair the cell's ability to maintain its appropriate cellular status. PMID- 14550748 TI - Induction of apoptosis in cultured human proximal tubule cells by fumonisins and fumonisin metabolites. AB - Fumonisin B1 (FB1) causes apoptosis in a variety of cell types and tissues but the apoptotic potential of other fumonisins and fumonisin metabolites has not been determined and the underlying mechanisms are not completely understood. In our studies we exposed human proximal tubule-derived cells (IHKE cells) to FB1, fumonisin B2 (FB2), fumonisin B3 (FB3), hydrolyzed fumonisin B1 (HFB1) and N palmitoyl-hydrolyzed fumonisin B1 (N-Pal-HFB1) and investigated caspase-3 activation, chromatin condensation and DNA fragmentation. Exposure to 10 micromol/L FB1 for 24 h led to a significant increase in caspase-3 activity, chromatin condensation and to DNA fragmentation. All other tested compounds did not show any significant activation of caspase-3 activity nor chromatin condensation and DNA-fragmentation. Furthermore, we examined if a sphinganine accumulation is correlated with an induction of apoptosis in IHKE cells. Therefore we used a liquid chromatography/electrospray ionization-mass spectrometry(LC/ESI-MS)-method using phytosphingosine as an internal standard to determine sphinganine and sphingosine concentrations in IHKE cells. Whereas a significant increase of sphinganine (up to 7000% compared to control cells) was observed with all fumonisin-derivates, sphingosine levels nearly remained unchanged indicating that all substrates inhibited ceramide synthase effectively. These results demonstrate that all compounds let to increased sphinganine levels in IHKE cells but only FB1 was able to induce apoptosis. We conclude that the inhibition of the ceramide synthase is not per se a predictor whether or not fumonisins induce apoptosis. PMID- 14550749 TI - Lansoprazole increases testosterone metabolism and clearance in male Sprague Dawley rats: implications for Leydig cell carcinogenesis. AB - Leydig cell tumours (LCTs) are frequently observed during rodent carcinogenicity studies, however, the significance of this effect to humans remains a matter of debate. Many chemicals that produce LCTs also induce hepatic cytochromes P450 (CYPs), but it is unknown whether these two phenomena are causally related. Our aim was to investigate the existence of a liver-testis axis wherein microsomal enzyme inducers enhance testosterone metabolic clearance, resulting in a drop in circulating hormone levels and a consequent hypertrophic response from the hypothalamic-pituitary-testis axis. Lansoprazole was selected as the model compound as it induces hepatic CYPs and produces LCTs in rats. Male Sprague Dawley rats were dosed with lansoprazole (150 mg/kg/day) or vehicle for 14 days. Lansoprazole treatment produced effects on the liver consistent with an enhanced metabolic capacity, including significant increases in relative liver weights, total microsomal CYP content, individual CYP protein levels, and enhanced CYP dependent testosterone metabolism in vitro. Following intravenous administration of [14C]testosterone, lansoprazole-treated rats exhibited a significantly smaller area under the curve and significantly higher plasma clearance. Significant reductions in plasma and testicular testosterone levels were observed, confirming the ability of this compound to perturb androgen homeostasis. No significant changes in plasma LH, FSH, or prolactin levels were detected under our experimental conditions. Lansoprazole treatment exerted no marked effects on testicular testosterone metabolism. In summary, lansoprazole treatment induced hepatic CYP-dependent testosterone metabolism in vitro and enhanced plasma clearance of radiolabelled testosterone in vivo. These effects may contribute to depletion of circulating testosterone levels and hence play a role in the mode of LCT induction in lansoprazole-treated rats. PMID- 14550750 TI - 3'-azido-3'-deoxythymidine (AZT) induces apoptosis and alters metabolic enzyme activity in human placenta. AB - The anti-HIV drug 3'-azido-3'-deoxythymidine (AZT) is the drug of choice for preventing maternal-fetal HIV transmission during pregnancy. Our aim was to assess the cytotoxic effects of AZT on human placenta in vitro. The mechanisms of AZT-induced effects were investigated using JEG-3 choriocarcinoma cells and primary explant cultures from term and first-trimester human placentas. Cytotoxicity measures included trypan blue exclusion, MTT, and reactive oxygen species (ROS) assays. Apoptosis was measured with an antibody specific to cleaved caspase-3 and by rescue of cells by the general caspase inhibitor Boc-D-FMK. The effect of AZT on the activities of glutathione-S-transferase, beta-glucuronidase, UDP-glucuronosyl transferase, cytochrome P450 (CYP) 1A, and CYP reductase (CYPR) in the placenta was assessed using biochemical assays and immunoblotting. AZT increased ROS levels, decreased cellular proliferation rates, was toxic to mitochondria, and initiated cell death by a caspase-dependent mechanism in the human placenta in vitro. In the absence of serum, the effects of AZT were amplified in all the models used. AZT also increased the amounts of activity of GST, beta-glucuronidase, and CYP1A, whereas UGT and CYPR were decreased. We conclude that AZT causes apoptosis in the placenta and alters metabolizing enzymes in human placental cells. These findings have implications for the safe administration of AZT in pregnancy with respect to the maintenance of integrity of the maternal-fetal barrier. PMID- 14550751 TI - Ah receptor regulation of mouse Cyp1B1 is additionally modulated by a second novel complex that forms at two AhR response elements. AB - Cyp1B1 is expressed constitutively in many extrahepatic cells and is induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). An enhancer region (AhER-810 to -1075 of the mouse Cyp1B1 promoter), which mediates aryl hydrocarbon receptor (AhR) regulation of transcription, contains three consensus XRE sequences (designated XRE1, XRE4, and XRE5) and a central Ebox. XRE5 is essential for both basal and induced activity in C3H10T1/2 cells. AhR/ARNT binding to XRE1, XRE4, and the Ebox complex function in combination to support the AhR/ARNT complex at XRE5. The identical 12 base cores of XRE1 and XRE4 differ from the core of XRE5 by two bases outside of the consensus XRE. These sites bind a constitutive complex slightly smaller than AhR/ARNT (anomalous complex; anC), which is not formed at XRE5 or six Cyp1A1 XREs. Exchange of these bases (m3 mutations) restores selective AhR/ARNT binding at XRE1/XRE4 and introduces anC binding at XRE5. The activities of multimeric XRE1 and XRE5 luciferase reporters responded in parallel to the extent of AhR/ARNT binding. The consensus anC binding sequence ((C/T)GCG(C/T)GCGC(C/A)GC) overlaps the XRE1/XRE4 AhR/ARNT element. Gel mobility analyses show that anC binds to XRE1/XRE4 under basal conditions, while AhR/ARNT partially displaces anC following TCDD induction. Selective depletion of anC with biotin-oligonucleotides increases AhR/ARNT binding. M3-mutations at, respectively, XRE1 and XRE4 of the AhER sequence, had opposite effects on luciferase reporters. Activities increased for the XRE1 mutation and decreased for the XRE4 mutation, but also depended on the level of AhR transfected into AhR -/- fibroblasts. AnC compete with AhR at XRE1 while playing an activating role at XRE4. This positive effect of constitutive anC binding at XRE4 may contribute to the characteristic basal Cyp1B1 expression in embryo fibroblasts, which is mediated by low constitutive activities of AhR. PMID- 14550752 TI - Role of glutathione redox cycle and catalase in defense against oxidative stress induced by endosulfan in adrenocortical cells of rainbow trout (Oncorhynchus mykiss). AB - The role of antioxidants in maintaining the functional integrity of adrenocortical cells during in vitro exposure to endosulfan, an organochlorine pesticide, was investigated in rainbow trout (Oncorhynchus mykiss). Aminotriazole (ATA), an inhibitor of catalase (CAT), L-buthionine sulfoximine (L-BSO), an inhibitor of glutathione (GSH) synthesis, and N-acetyl cysteine (NAC), a glutathione precursor, were used to investigate the role of CAT and GSH redox cycle in protection against the adrenal toxicity of endosulfan, a pesticide that impairs cell viability (LC50 366 microM) and cortisol secretion (EC50 19 microM) in a concentration-related manner. Pretreatment with ATA and L-BSO enhanced the toxicity of endosulfan (LC50 and EC50, respectively, 302 and 2.6 microM with ATA, 346 and 3.1 microM with L-BSO), while pretreatment with NAC had no significant effect on cell viability and increased the EC50 of endosulfan to 51 microM. CAT activity was significantly reduced following exposure to endosulfan when cells were pretreated with ATA. Pretreatment with L-BSO significantly decreased glutathione peroxidase (GPx) activity and reduced glutathione (GSH) levels in a concentration-related manner following exposure to endosulfan, while GSH levels were significantly higher in NAC pretreated cells compared to untreated cells. Finally, pretreatment with ATA and L-BSO increased, while pretreatment with NAC decreased, lipid hydroperoxides (LOOH) levels. CAT, GPx, and GSH were identified as important antioxidants in maintaining the function and integrity of rainbow trout adrenocortical cells and ATA, L-BSO, and NAC were identified as effective modulators of CAT and GSH redox cycle. Moreover, this study suggests that the glutathione redox cycle may be more efficient than catalase in protecting adrenocortical cells against endosulfan-induced oxidative stress. PMID- 14550753 TI - Regulation of occupational exposures in China. AB - The recent passage of the Occupational Diseases Prevention and Control Act of 2002 (ODPCAct) in China and the new occupational exposure limits signify the Chinese government's commitment to improve the environment of the workplace and to eradicate preventable occupational diseases. The effectiveness of the ODPCAct, however, will depend on not only implementation and enforcement but also education and communication. For large industrial facilities, implementation of the new regulations can be enforced with periodic monitoring and inspections. The difficulty will come from small makeshift or crudely converted workshops in villages and small towns in rural areas. The challenge will be to reach out to these small workshop owners and workers, i.e., to communicate and inform them about the newly promulgated regulations, the business owners' legal responsibility and liability, and the workers' right to a safe workplace. Attention and resources should be focused on educating both shop owners and workers about the hazards of the chemicals that they use, basic requirements for a safe workplace, preventive measures, and controls to reduce exposures. PMID- 14550754 TI - New trends in the development of occupational exposure limits for airborne chemicals in China. AB - Occupational exposure limits (OELs) are well established in many countries, which serve occupational professionals as benchmarks of industrial hygiene practice at workplaces worldwide. Starting in the mid-1950s, the central government of China began promulgating OELs for hazardous substances at workplaces. This paper discusses the historical basis, philosophical principles and schematic protocols of developing and setting OELs in China. The underlying principles include: (1) protection of human health being the first and the most important criterion; (2) the use of quantitative epidemiological studies in humans being given top priority; (3) integration and full use of all information sources, including animal experimental data for new chemicals or chemicals with new toxicity concerns; (4) considerations of socioeconomic and technological feasibilities in the country; and (5) amending existing standards based on new evidence. The strategy of the World Health Organization's "Two-step Procedure" is applied to convert health-based recommendations to law-based operational OELs, with considerations for national technological and socioeconomic conditions and priorities. As a result of the recent passage of the new law Occupational Diseases Prevention and Control Act of the People's Republic of China (ODPCAct), an official document Occupational Exposure Limits for Hazardous Agents in the Workplace containing a comprehensive list of new and amended OELs has been issued, which has now become one of the most essential regulations affiliated with the ODPCAct. This paper provides a brief summary of the salient features of the new law ODPCAct and the principles and processes of developing or amending OELs. This paper also discusses the challenges that lie ahead in enforcing the new regulations in China. PMID- 14550755 TI - Food consumption and body weight changes with neotame, a new sweetener with intense taste: differentiating effects of palatability from toxicity in dietary safety studies. AB - Safety studies done with neotame, a sweetener with intense taste, demonstrate that changes in bodyweight (BW) and BW gain (BWG) are due to reduced food consumption (FC) rather than toxicity. When offered a choice, rats preferred basal diet to diet with relatively low concentrations of neotame. When no choice was available, rats ate less as concentrations increased, demonstrating reduced palatability. Changes in dietary concentrations of neotame resulted in changes in FC. The maximum tolerable doses (MTDs) in rats, dogs, and mice were due to decreases in BWG secondary to poor palatability of diets when neotame concentrations exceeded approximately 35,000 ppm. Concentrations were increased as animals grew to maintain constant dosing on a "mg/kg bw/day" basis. Food conversion efficiency (FCE) was not changed in rats during periods of active growth. The only consistent findings across safety studies were reductions in BW, BWG, and FC with no dose-response in rats, mice, and dogs. In definitive safety studies, there were no adverse findings related to neotame treatment from clinical observations, physical examinations, water consumption, or clinical pathology evaluations; nor was there morbidity, mortality, organ toxicity, macroscopic or microscopic postmortem findings. Analysis of data from long-term studies in Sprague-Dawley rats support the conclusion that changes in FC alone can cause the observed changes in BWG in neotame studies when changes are scaled allometrically [Regul. Toxicol. Pharmacol. (2003)]. Consequently, BW parameters are not appropriate endpoints for setting no-observed-effect levels (NOELs) for neotame. PMID- 14550756 TI - Long-term food consumption and body weight changes in neotame safety studies are consistent with the allometric relationship observed for other sweeteners and during dietary restrictions. AB - In long-term safety studies with neotame, a new high-intensity sweetener 7000 13,000 times sweeter than sucrose, the percent changes (%Delta) in body weight gain (BWG) in Sprague-Dawley rats were several-fold greater than the %Delta in overall food consumption (FC). This study investigates the question of whether the changes in BWG were adverse or secondary to small, long-term decrements in FC. The hypothesis tested in Sprague-Dawley rats was that the relationship between long-term %Delta in FC and %Delta in BWG is linear and in a ratio of 1:1. The %Delta in FC were compared to %Delta in BWG after 52 weeks on study in one saccharin (825 rats), two sucralose (480 rats), two neotame (630 rats), and five dietary restriction (>1000 rats) studies. Non-transformed plotting of data points demonstrated an absence of linearity between %Delta in FC and %Delta in BWG; however, log-log evaluation demonstrated a robust (R2=0.97) linear relationship between %Delta in FC and %Delta in BWG. This relationship followed the well-known allometric equation, y=bxa where x is %DeltaFC, y is %DeltaBWG, b is %DeltaBWG when DeltaFC=1, and a is the log-log slope. Thus, in Sprague-Dawley rats at week 52, the long-term relationship between %Delta in FC and %Delta in BWG was determined to be: %DeltaBWG=3.45(%DeltaFC0.74) for males and %DeltaBWG=5.28(%DeltaFC0.68) for females. Sexes were statistically different but study types, i.e., the high-intensity sweeteners saccharin and sucralose versus dietary restriction, were not. The %Delta in BWG are allometrically consistent with the observed %Delta in FC for these high-intensity sweeteners, including neotame. BW parameters are not appropriate endpoints for setting no-observed effect levels (NOELs) when materials with intense taste are admixed into food. An approach using objective criteria is proposed to delineate BW changes due to toxicity from those secondary to reduced FC. PMID- 14550757 TI - Procedures for calculating cessation lag. AB - Environmental regulations aimed at reducing cancer risks usually have the effect of reducing exposure to a carcinogen at the time the regulation is implemented. The reduction of cancer risk may occur shortly after the reduced exposure or after a considerable period of time. The time of risk reduction associated with exposure reduction will vary by compound. Some recommended measures of the economic benefits associated with environmental regulations are sensitive to the timing of the risk reductions and cannot be effectively addressed by the conventional dose-response procedures. This paper introduces the concept and methodologies for calculating cessation lag effects, with the specific goal of answering the following questions: (1) How many cancer cases are avoided at age t after cessation (or reduction) of exposure concentration? and (2) How long does the effect of an exposure last after exposure has terminated (or been reduced)? The proposed procedures do not require more information than what is required by the conventional dose-response procedures for which cumulative or an averaged lifetime exposure is used. PMID- 14550758 TI - Local tolerance testing of parenteral drugs: how to put into practice. AB - Notwithstanding that there are national and international guidelines about local tolerance testing of parenteral drugs in animals, in particular to mention CPMP/SWP/2145/00 (Note for Guidance on Non-Clinical Local Tolerance Testing of Medicinal Products), very heterogeneous study designs have been established in the past. A working group including experts of the leading pharmaceutical industry from German-language countries, named "Arbeitskreis Lokale Vertraglichkeit," has been intensively discussing the experimental procedures in detail for a period of six years and has been considering their pros and cons. This team of experts now feels confident to give some recommendations for study conduct besides describing different materials and methods for this type of toxicological study. Special knowledge from toxicologists as well as pathologists from our working group has been taken into account. This paper deals with choice of species, number of animals used, controls, administration sites, volumes, rate and frequency, length of observation period, termination, clinical, macroscopic and histopathological examinations and, finally, overall assessment criteria and conclusion. Our purpose is that this paper may be of value for: *The study director who is inexperienced in the conduction of local tolerance testing and who may need a standard design as his first step into this new field. *The well versed study director who would like to know how others have done in the past, who may examine self-critically his own practice and who is open to our team's recommendations, tips and tricks from practice. *The specialist at a regulatory authority who, finally, reviews study reports, assesses their format and content and, above all, decides on the approval of a drug product. PMID- 14550759 TI - An updated evaluation of the carcinogenic potential of 1,4-dioxane. AB - This paper presents a critical review of the information pertaining to the potential carcinogenicity of 1,4-dioxane. The primary target organs for cancer via the oral route are the liver and the nasal cavity, however, the relevance of nasal cavity tumors to human exposures has been questioned. Liver tumors were accompanied by degenerative changes and appear only to occur at high doses where clearance mechanisms are saturated and liver toxicity is significant. Genetic toxicity data suggests that 1,4-dioxane is a very weak genotoxin. An increase in hepatocyte cell proliferation was reported and 1,4-dioxane was shown to act as a tumor promoter in rat liver and mouse skin carcinogenicity assays. Two reports are available from the literature regarding physiologically based pharmacokinetic (PBPK) modeling approaches to assess the risk of liver cancer for 1,4-dioxane. A comparison of cancer risk estimates from linear and nonlinear models in the presence or absence of PBPK modeling suggests that USEPAs current cancer slope factor significantly overestimates the potential cancer risk from 1,4-dioxane. This critical review of the scientific literature indicates that a formal reevaluation of the carcinogenic potency of 1,4-dioxane is warranted. PMID- 14550760 TI - A model to estimate the oestrogen receptor mediated effects from exposure to soy isoflavones in food. AB - The advantages that regular consumption of a diet containing soy may have on human health have been enshrined in a major health claim that has been approved by the Food and Drug Administration in the USA, regarding potential protection from heart disease by soy. This could have a major influence on the dietary consumption patterns of soy for consumers and lead to the development of soy enriched foods to enable consumers to achieve the benefits thought to be associated with increased soy consumption in a Western diet. If an increase in soy consumption is beneficial to particular disease conditions, there is always the possibility that there will be effects other than those that are desirable. For soy-containing foods there has been concern that the phytoestrogen content of soy, which is composed of several isoflavones, could be a separate health issue, due to the oestrogen-like activity of isoflavones. To address this, a method has been developed to estimate, relative to 17-beta oestradiol, the activity of the common isoflavones present in soy phytoestrogens, based on their binding to and transcriptional activation of the major oestrogen receptor sub-types alpha and beta. Using this approach, the additional oestrogen-like activity that would be expected from inclusion of soy supplemented foodstuffs in a Western diet, can be determined for different sub-populations, who may have different susceptibilities to the potential for the unwanted biological effects occurring with consumption of soy enriched foods. Because of the theoretical nature of this model, and the controversy over the nature of whether some of the oestrogen-like effects of phytoestrogens are adverse, the biological effects of soy isoflavones and their potential for adverse effects in man, is also reviewed. The question that is critical to the long term safe use of foods enriched in soy is, which observed biological effects in animal studies are likely to also occur in man and whether these would have an adverse effect on human health. PMID- 14550761 TI - Review of procedures for protecting human subjects in recent clinical studies of pesticides. AB - Arguments have been made for and against the regulatory use of data from human subjects on both scientific and ethical grounds. One argument against the use of data from human clinical studies involving pesticides asserts that such data are obtained from studies that do not follow the Common Rule (40 CFR 26), which provides procedures for protecting human subjects in studies funded by federal agencies, including the U.S. Environmental Protection Agency (U.S. EPA). Although privately conducted studies using human subjects are not legally subject to or required to comply with the Common Rule, the protections of the Declaration of Helsinki and the International Conference on Harmonisation (ICH) Good Clinical Practice are commonly followed. We sought to answer the question of whether recent human clinical studies with insecticides performed according to Good Clinical Practice provided volunteers with the same protections as the Common Rule. All three sets of guidance have in common the intent to protect volunteer human subjects by providing standards for the conduct of studies in which they participate. This analysis compares the elements of the Common Rule with comparable elements from the Declaration of Helsinki and Good Clinical Practice to evaluate similarities and differences in procedural requirements. It then evaluates the documentation from 15 recent human studies of twelve insecticides conducted at four clinical laboratories in order to determine whether the conduct of those studies is consistent with the protections of the Common Rule. There were some cases for which we could not verify compliance with certain Common Rule elements; however, based on our evaluation it is apparent that the studies we reviewed were conducted in a manner substantially consistent with the fundamental protections of the Common Rule-voluntary participation, informed consent, and review by an ethical committee or institutional review board. PMID- 14550762 TI - A proposed inhalation reference concentration for methanol. AB - A biologically based approach was taken to developing an inhalation Reference Concentration (RfC) for methanol, a high production volume chemical with many commercial applications, including use as an alternative fuel for motor vehicles and as a hydrogen source for fuel cells. Benchmark Dose methodology was applied to the most sensitive toxic endpoint for assessing potential health risks in humans, cervical rib malformation data obtained using CD-1 mice. The concentration of methanol in circulating blood was employed as the dose metric, and the maximum likelihood estimate of the blood methanol increment causing a 10% extra risk of these malformations, was 215.4 mg/L, with a lower 95% confidence bound of 97.4 mg/L. A "Reference Increment" for blood methanol was then determined by dividing this value by a 3-fold factor for residual pharmacodynamic uncertainty between species and a 10-fold factor for interindividual variation in human sensitivity to methanol. The resulting Reference Increment in blood methanol was then converted to an equivalent inhalation Reference Concentration with a physiologically based pharmacokinetic model evaluated for continuous exposure conditions. The resulting maximum likelihood estimate for the inhalation RfC was 298 mg/m3, with a 95% lower confidence bound of 135 mg/m3. PMID- 14550763 TI - Trace element risk assessment: essentiality vs. toxicity. AB - Risk assessment of essential trace elements examines high intakes resulting in toxicity and low intakes resulting in nutritional deficiencies. This paper analyzes the risk assessments carried out by several U.S. governmental and private organizations for eight essential trace elements: chromium, copper, iodine, iron, manganese, molybdenum, selenium, and zinc. The compatibility of the toxicity values with the nutritionally essential values is examined, in light of recently derived values, termed Dietary Reference Intakes, set by the U.S. Food and Nutrition Board of the Institute of Medicine. The results show that although there are differences in the values set by the different organizations, increased coordination has resulted in values that are more compatible than revealed in past evaluations. PMID- 14550764 TI - Cyclic AMP and tumor necrosis factor-alpha regulate CXCR4 gene expression in Schwann cells. AB - Rat peripheral nerve Schwann cells have been shown to express the alpha-chemokine receptor CXCR4 as well as the corresponding ligand stromal cell-derived factor-1 (SDF-1). We have investigated gene regulatory mechanisms acting on the expression of CXCR4 in cultured rat Schwann cells and found that receptor expression at transcript- and protein levels is directly dependent on intracellular cyclic AMP. Such increased levels of CXCR4 expression were found to be efficiently reversed by the action of tumor necrosis factor-alpha (TNFalpha). We also provide evidence that the POU box transcription factor Oct-6/SCIP is involved in the control of CXCR4 transcription. Finally, we could demonstrate that CXCR4 activation by SDF 1alpha increases the number of dying Schwann cells, indicating that this receptor/ligand interaction is modulating cell survival. Our data, therefore, suggest that in the Schwann cell lineage signal transduction cascades controlled by the activation of TNF- and CXCR4 receptors are functionally coupled. PMID- 14550765 TI - Specific involvement in neuropathic pain of AMPA receptors and adapter proteins for the GluR2 subunit. AB - Chronic pain states arise from peripheral nerve injury and are inadequately treated with current analgesics. Using intrathecal drug administration in a rat model of neuropathic pain, we demonstrate that AMPA receptors play a role in the central sensitisation that is thought to underpin chronic pain. The GluR2 subunit of the AMPA receptor binds to a number of intracellular adapter proteins including GRIP, PICK1 and NSF, which may link the receptor to proteins with signalling, scaffolding and other roles. We implicate for the first time a possible role for GRIP, PICK1 and NSF in neuropathic sensitisation from experiments with cell-permeable blocking peptides mimicking their GluR2 interaction motifs and also demonstrate differential changes in expression of these proteins following peripheral nerve injury. These studies suggest a critical involvement of protein:protein complexes associated with the AMPA receptor in neuropathic pain, and the possibility that they may have potential as novel therapeutic targets. PMID- 14550766 TI - IGF-I has a direct proliferative effect in adult hippocampal progenitor cells. AB - The aim of the present study was to investigate the potential direct effects of insulin-like growth factor-I (IGF-I) on adult rat hippocampal stem/progenitor cells (AHPs). IGF-I-treated cultures showed a dose-dependent increase in thymidine incorporation, total number of cells, and number of cells entering the mitosis phase. Pretreatment with fibroblast growth factor-2 (FGF-2) increased the IGF-I receptor (IGF-IR) expression, and both FGF-2 and IGF-I were required for maximal proliferation. Time-lapse recordings showed that IGF-I at 100 ng/ml decreased differentiation and increased proliferation of single AHPs. Specific inhibition of mitogen-activated protein kinase kinase (MAPKK), phosphatidylinositol 3-kinase (PI3-K), or the downstream effector of the PI3-K pathway, serine/threonine p70 S6 kinase (p70(S6K)), showed that both the MAPK and the PI3-K pathways participate in IGF-I-induced proliferation but that the MAPK activation is obligatory. These results were confirmed with dominant-negative constructs for these pathways. Stimulation of differentiation was found at a low dose (1 ng/ml) of IGF-I, clonal analysis indicating an instructive component of IGF-I signaling. PMID- 14550767 TI - GDNF increases the survival of developing oculomotor neurons through a target derived mechanism. AB - Glial cell line-derived neurotrophic factor (GDNF) is the most potent motoneuron survival factor. We show here that in the chick oculomotor system, endogenous GDNF is derived largely from extraocular muscle but less from glial cells and not from muscle spindles. Increased levels of GDNF exclusively in the target rescued 30% of oculomotor neurons that would normally die during developmental cell death, a rate of rescue similar to that with systemic GDNF application. Thus, GDNF supports motoneuron survival in a retrograde, target-derived fashion, as opposed to a local paracrine route or an indirect route via sensory afferents. Persephin, another member of the GDNF family, did not increase survival with target delivery, despite its retrograde transport from the target. Unlike GDNF, however, persephin increased neurite outgrowth from oculomotor nuclei in vitro. Thus, one GDNF family member acts as a muscle-derived retrograde survival factor, whereas another one has distinct functions on neurite outgrowth. PMID- 14550768 TI - Essential role of mitochondrial permeability transition in vanilloid receptor 1 dependent cell death of sensory neurons. AB - Capsaicin causes pain by activating VR1, a cloned capsaicin receptor, in sensory neurons. After the initial excitatory responses, capsaicin produces prolonged analgesia, presumably because of the neurotoxic effect that leads to the death of sensory neurons. However, the mechanism underlying capsaicin-induced cell death of sensory neurons is not known. Here we report that capsaicin induces cell death in VR1-expressing sensory neurons and VR1-transfected human embryonic kidney cells. Cell death of sensory neurons induced by capsaicin is accompanied by DNA fragmentation, TUNEL staining, and shrinkage of the nucleus in a caspase dependent manner, indicating the apoptotic nature of the cell death. Mitochondrial permeability transition is likely to be a major component of capsaicin-induced cell death because bonkrekic acid and cyclosporin A, inhibitors of mitochondrial permeability transition, block this cell death. These results imply that capsaicin induces mitochondrial dysfunction in VR1-expressing cells, leading to apoptotic cell death, which is a well-known neurotoxic effect of capsaicin. PMID- 14550769 TI - Roles of STEF/Tiam1, guanine nucleotide exchange factors for Rac1, in regulation of growth cone morphology. AB - Rho family GTPases are suggested to be pivotal for growth cone behavior, but regulation of their activities in response to environmental cues remains elusive. Here, we describe roles of STEF and Tiam1, guanine nucleotide exchange factors for Rac1, in neurite growth and growth cone remodeling. We reveal that, in primary hippocampal neurons, STEF/Tiam1 are localized within growth cones and essential for formation of growth cone lamellipodia, eventually contributing to neurite growth. Furthermore, experiments using a dominant-negative form demonstrate that STEF/Tiam1 mediate extracellular laminin signals to activate Rac1, promoting neurite growth in N1E-115 neuroblastoma cells. STEF/Tiam1 are revealed to mediate Cdc42 signal to activate Rac1 during lamellipodial formation. We also show that RhoA inhibits the STEF/Tiam1-Rac1 pathway. These data are used to propose a model that extracellular and intracellular information is integrated by STEF/Tiam1 to modulate the balance of Rho GTPase activities in the growth cone and, consequently, to control growth cone behavior. PMID- 14550770 TI - cAMP-dependent protein kinase A mediation of vasopressin gene expression in the hypothalamus of the osmotically challenged rat. AB - We have tested the hypothesis that 3', 5'-cyclic adenosine monophosphate (cAMP) dependent protein kinase A (PKA) is involved in the regulation of the vasopressin (VP) gene in the magnocellular neurons of the paraventricular nucleus (PVN) of the osmotically challenged rat. An adenoviral vector expressing a potent peptide inhibitor of PKA, Ad.CMV.PKIalpha, was demonstrated to be highly efficient in vitro. Ad.CMV.PKIalpha was then introduced into the PVN of rats bearing a VP reporter transgene (3-VCAT-3) consisting of the VP structural gene containing an epitope reporter in exon III, flanked by 3 kb of upstream and 3 kb of downstream sequence Robust transgene expression is seen in VP neurons of the PVN, and this increases following 72 h of dehydration. Ad.CMV.PKIalpha significantly blunted 3 VCAT-3 expression in the osmotically stimulated PVN. Our evidence suggests that PKA mediates changes in VP gene expression in response to dehydration through sequences contained within the 3-VCAT-3 transgene. PMID- 14550771 TI - Mutations in the lipid-binding domain of alpha-synuclein confer overlapping, yet distinct, functional properties in the regulation of dopamine transporter activity. AB - Alpha-synuclein and its missense mutants (A30P, A53T) have been linked to the genesis of idiopathic and rare familial forms of Parkinson's disease, respectively. Here we show that, similar to the wild-type alpha-synuclein, the A30P mutant forms a strong complex with the human dopamine transporter (hDAT), through direct protein:protein interactions between the nonamyloid beta component (NAC) domain of the A30P mutant and the last 22 aminoacyl residues of the carboxy terminal tail of hDAT. The A30P mutant negatively modulates hDAT functional activity and to a greater extent than wild-type alpha-synuclein, with reduced uptake of extracellular dopamine and dopamine-mediated, hDAT-dependent cytotoxicity. By contrast, the A53T mutant neither forms a strong protein:protein complex with hDAT nor modulates dopamine uptake by hDAT, and dopamine-mediated, hDAT-dependent cytotoxicity is higher than with either wild-type or the A30P variant of alpha-synuclein, but not significantly different from that of cells expressing hDAT alone. Confocal microscopy shows substantial overlap in colocalization of all three alpha-synuclein variants with hDAT, with only minor differences. Although the complex formation with hDAT occurs through the NAC domain of the alpha-synuclein variants, it is the familial Parkinson's disease linked missense mutations present in the amino-terminal lipid binding domain of the alpha-synuclein variants that dictate the extent of the regulation of hDAT function. These studies highlight previously unknown properties of the A30P and the A53T mutants of alpha-synuclein with respect to the modulation of hDAT activity and/or regulation, and its subsequent functional outcome, which are uniquely distinct. PMID- 14550772 TI - High basal expression and injury-induced down regulation of two regulator of G protein signaling transcripts, RGS3 and RGS4 in primary sensory neurons. AB - The regulators of G-protein signaling (RGS) proteins are a family of intracellular modulators of G-protein coupled receptor (GPCR) sensitivity. They act as GTPase accelerating proteins returning the Galpha subunit back to an inactive latent state. We find that RGS3 and RGS4 are constitutively expressed at high levels in C-fiber primary sensory neurons in the adult rat dorsal root ganglion (DRG) and transection of the sciatic nerve results in a substantial down regulation of these transcripts. RGS4 mRNA is expressed only in GDNF-responsive neurons and GDNF supports the expression of this transcript in primary DRG cultures. A PDZ domain containing subtype of RGS3 is the most abundant and regulated form of this protein within the DRG. Decreased levels of RGS3 and RGS4 in injured sensory neurons is likely to result in an increased GPCR sensitivity, and therefore contribute to alterations in cellular function seen after such lesions. PMID- 14550773 TI - Identification and characterization of Slitrk, a novel neuronal transmembrane protein family controlling neurite outgrowth. AB - The Slitrk family consists of six structurally related transmembrane proteins (Slitrk1-6) in the mouse. In the extracellular region, they share two conserved leucine-rich repeat domains that have a significant homology to a secreted axonal growth-controlling protein, Slit. These proteins also have a homology to trk neurotrophin receptors in their intracellular domains, sharing a conserved tyrosine residue. Expression of Slitrk is highly restricted to neural tissues, but varies within the family. More specifically, Slitrk1 expression is in the mature neurons, whereas Slitrk2 is strongly expressed in the ventricular layer, and Slitrk6 shows compartmentalized expression in diencephalon. Over-expressed Slitrk1 induced unipolar neurites in cultured neuronal cells, whereas Slitrk2 and other Slitrk proteins inhibited neurite outgrowth. Deletion analysis showed that the functional difference between Slitrk1 and Slitrk2 lies in their intracellular domains, which are conserved in Slitrk2-6, but not in Slitrk1. These results suggest that the Slitrk proteins are the neuronal components that control the neurite outgrowth. PMID- 14550774 TI - Ciliary neurotrophic factor is an early lesion-induced retrograde signal for axotomized facial motoneurons. AB - To investigate the involvement of ciliary neurotropic factor (CNTF) in the postlesional response of motoneurons, we studied the activation of STAT3 signaling, the main signal transduction pathway of CNTF-like cytokines, in the facial nucleus of wildtype and CNTF-deficient mice following peripheral nerve transection. As shown by immunocytochemistry and immunoblot analysis, phosphorylation and nuclear translocation of STAT3 was maximally induced within 12 h postlesion in motoneurons of the ipsilateral facial nucleus of wildtype mice and is maintained for at least 3 days. In CNTF(-/-) mouse mutants, activation of STAT3 signaling was delayed by 10-12 h. Application of CNTF to the transected nerve restored rapid STAT3 activation in CNTF-deficient animals, whereas application of colchicine suppressed STAT3 signaling in wildtype mice for at least 24 h. These results identify CNTF as an early retrograde signal in axotomized facial motoneurons by showing that CNTF released at the lesion site is responsible for the initial induction of STAT3 signaling. Other cytokines like leukemia inhibitory factor obviously become active at later time points. PMID- 14550775 TI - Targeting conditional gene modification into the serotonin neurons of the dorsal raphe nucleus by viral delivery of the Cre recombinase. AB - Delivery of viral vectors encoding the Cre recombinase is showing promise to target gene modification in specific brain regions. Here we describe the targeting of the dorsal raphe nucleus (DRN), which contains the majority of the serotonin (5-HT) neurons projecting to the forebrain. First, we demonstrate successful transgene expression in the mouse DRN by stereotaxic delivery of the AdnlslacZ adenoviral vector. Second, we show that expression of the Cre recombinase can be achieved in the 5-HT neurons by optimized injection of the Adcre vector. Using reporter mice in which Cre activity induces beta galactosidase (beta-gal) expression, we demonstrate efficient Cre-mediated recombination and persistence of beta-gal positive 5-HT neurons at least 1 month postinjection. Together, these results demonstrate that viral delivery provides a valuable method to target Cre recombination throughout the murine DRN and thus to study 5-HT neurotransmission by conditional gene modification. PMID- 14550776 TI - Molecular cloning of Bral2, a novel brain-specific link protein, and immunohistochemical colocalization with brevican in perineuronal nets. AB - The hyaluronan binding chondroitin sulphate proteoglycans, called lecticans, are the abundant extracellular matrix molecules in the developing and/or adult brain. The link proteins (LPs) are also known to be coordinately present in brain. We report here the molecular cloning and expression analysis of a novel member of LPs: Bral2, predominantly expressed in brain. The Bral2 mRNA expression is first detected at P20 and continued through adulthood, suggesting its functional importance and association with adult-type lecticans. The substantial immunoreactivity of Bral2 is found in several nuclei throughout the midbrain and hindbrain in a perineuronal net pattern. In situ hybridization revealed that Bral2 is synthesized by these neurons themselves, especially by the GABAergic neurons in the cerebellar cortex. Interestingly, the colocalization and synergic importance of Bral2 and brevican in the perineuronal nets is indicated by the comparative immunohistochemical analysis using wild-type and brevican-deficient mouse brain. Our results suggest that Bral2 is involved in the formation of extracellular matrix contributing to perineuronal nets and facilitate the understanding of a functional role of these extracellular matrices. PMID- 14550777 TI - Neural crest stem cell and cardiac endothelium defects in the TrkC null mouse. AB - TrkC null mice have multiple cardiac malformations. Since neural crest cells participate in cardiac outflow tract septation, the aim of this study was to determine at the cellular level the putative neural crest defect. We have identified three types of progenitor cells: stem cells that undergo self-renewal and can generate many cell types, cells that are restricted in their developmental potentials, and cells that are committed to the smooth muscle cell lineage. In TrkC null mice, there is a greater than 50% decrease in stem cell numbers and an equivalent increase in fate-restricted cells. The outflow tract wall is thickened and the endothelial tube is disorganized. We conclude that deletion of the TrkC gene causes precocious fate restrictions of the neural crest stem cell and a defect of the outflow tract endothelium, both of which may contribute to the outflow tract malformations that occur in TrkC null mice. PMID- 14550778 TI - Directed migration of neuronal precursors into the ischemic cerebral cortex and striatum. AB - Pathological processes, including cerebral ischemia, can enhance neurogenesis in the adult brain, but the fate of the newborn neurons that are produced and their role in brain repair are obscure. To determine if ischemia-induced neuronal proliferation is associated with migration of nascent neurons toward ischemic lesions, we mapped the migration of cells labeled by cell proliferation markers and antibodies against neuronal marker proteins, for up to 2 weeks after a 90-min episode of focal cerebral ischemia caused by occlusion of the middle cerebral artery. Doublecortin-immunoreactive cells in the rostral subventricular zone, but not the dentate gyrus, migrated into the ischemic penumbra of the adjacent striatum and, via the rostral migratory stream and lateral cortical stream, into the penumbra of ischemic cortex. These results indicate that after cerebral ischemia, new neurons are directed toward sites of brain injury, where they might be in a position to participate in brain repair and functional recovery. PMID- 14550779 TI - Hepatocyte growth factor promotes proliferation and neuronal differentiation of neural stem cells from mouse embryos. AB - Hepatocyte growth factor (HGF), originally cloned as a hepatocyte mitogen, has recently been reported to exhibit neurotrophic activity in addition to being expressed in different parts of the nervous system. At present, the effects of HGF on neural stem cells (NSCs) are not known. In this study, we first report the promoting effect of HGF on the proliferation of neurospheres and neuronal differentiation of NSCs. Medium containing only HGF was capable of inducing neurosphere formation. Addition of HGF to medium containing fibroblast growth factor 2 or epidermal growth factor increased both the size and number of newly formed neurospheres. More neurons were also obtained when HGF was added in differentiation medium. In contrast, neurosphere numbers were reduced after repeated subculture by mechanical dissociation, suggesting that HGF-formed neurospheres comprised predominantly progenitor cells committed to neuronal or glial lines. Together, these results suggest that HGF promotes proliferation of neurospheres and neuronal differentiation of NSCs derived from mouse embyos. PMID- 14550780 TI - The human NTERA2 neural cell line generates neurons on growth under neural stem cell conditions and exhibits characteristics of radial glial cells. AB - NTERA2 cells are a human neural cell line generating neurons after exposure to retinoic acid and, as such, are widely used as a model of neurogenesis. We report that these cells form spheres when grown in serum-free medium supplemented with basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF). These spheres were found to express markers of radial glial cells such as, Pax6, glutamate transporter (GLAST), tenascin C, brain lipid-binding protein (BLBP), and the 3CB2 antigen. On plating on an adhesive substrate, NTERA2 spheres generate a large percentage of immature neurons (30-50%) together with a minority of cells of the oligodendrocyte lineage. Thus NTERA2 cells share properties with neural stem cells. However, at variance with the latter, we found that they produce their own bFGF implicated in an autocrine or paracrine proliferative loop and that they do not generate astrocytes after differentiation. These results provide an interesting model to study radial glial cells and their role in human neurogenesis. PMID- 14550781 TI - GABA B receptor subunit expression in glia. AB - GABA(B) receptor subunits are widely expressed on neurons throughout the CNS, at both pre- and postsynaptic sites, where they mediate the late, slow component of the inhibitory response to the major inhibitory neurotransmitter GABA. The existence of functional GABA(B) receptors on nonneuronal cells has been reported previously, although the molecular composition of these receptors has not yet been described. Here we demonstrate for the first time, using immunohistochemistry the expression of GABA(B1a), GABA(B1b), and GABA(B2) on nonneuronal cells of the rat CNS. All three principle GABA(B) receptor subunits were expressed on these cells irrespective of whether they had been cultured or found within brain tissue sections. At the ultrastructural level GABA(B) receptor subunits were expressed on astrocytic processes surrounding both symmetrical and assymetrical synapses in the CA1 subregion of the hippocampus. In addition, GABA(B1a), GABA(B1b), and GABA(B2) receptor subunits were expressed on activated microglia in culture but were not found on myelin forming oligodendrocytes in the white matter of rat spinal cord. Together these data demonstrate that the obligate subunits of functional GABA(B) receptors are expressed in astrocytes and microglia in the rat CNS. PMID- 14550782 TI - HIV-1 Tat protein upregulates inflammatory mediators and induces monocyte invasion into the brain. AB - Impaired inflammatory functions may be critical factors in the mechanisms by which HIV-1 enters the CNS. Evidence indicates that a viral gene product, the protein Tat, can markedly contribute to these effects. In the present study we tested the hypothesis that Tat can upregulate the expression of inflammatory cytokines and adhesion molecules and facilitate the entry of monocytes into the brain. Expression of inflammatory mediators such as monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-alpha (TNF-alpha), vascular cell adhesion molecule-1 (VCAM-1), and intercellular adhesion molecule-1 (ICAM-1) was assessed in C57BL/6 mice injected with Tat(1-72) into the right hippocampus. In the Tat(1-72)-injected groups, mRNA and protein levels of MCP-1, TNF-alpha, VCAM 1, and ICAM-1 were markedly elevated compared to those in control animals. The most pronounced changes were observed in and around the injected hippocampus. Double-labeling immunohistochemistry demonstrated that inflammatory proteins were primarily expressed in activated microglial cells and perivascular cells. In addition, astrocytes and endothelial cells were susceptible to Tat(1-72)-induced inflammatory responses. These changes were associated with a substantial infiltration of monocytes into the brain. These data demonstrate that intracerebral administration of Tat can induce profound proinflammatory effects in the brain, leading to monocyte infiltration. PMID- 14550783 TI - The concerted signaling of ERK1/2 and JNKs is essential for PC12 cell neuritogenesis and converges at the level of target proteins. AB - Mitogen-activated protein kinase (MAPK) pathways are central signaling elements, which translate and integrate stimuli from cell surface receptors into cytoplasmic and transcriptional responses. Here, we systematically compare the role of MAPKs in the nerve growth factor-induced long-term differentiation of PC12 cells and show the persistent nuclear and dose-dependent cytoplasmic activation of extracellular signal-regulated kinases 1 and 2 (ERK1/2) and the increasing nuclear and cytoplasmic activation of c-Jun N-terminal kinases (JNKs). Inhibition of ERK1/2 and JNKs significantly reduced neurite outgrowth. Both synergistically controlled the expression of c-Jun, the induction and/or phosphorylation of neurofilament, and the phosphorylation of Elk-1. JNKs alone were responsible for the phosphorylation of c-Jun and activating transcription factor 2 as well as for the expression of MAPK phosphatase 1. In contrast, p38alpha was only transiently activated and marginally involved in these processes. Thus, JNKs and ERK1/2 accomplish differentiation by signaling in parallel cascades that converge only at the target level. PMID- 14550784 TI - Extrasynaptic GABA(A) channels activated by THIP are modulated by diazepam in CA1 pyramidal neurons in the rat brain hippocampal slice. AB - Single-channel currents were activated by THIP (4,5,6,7-tetrahydroisoxazolo[5,4 c]pyridin-3-ol) in cell-attached patches on CA1 pyramidal neurons in the rat hippocampal slice preparation. THIP activated GABA(A) channels after a delay that was concentration-dependent and decreased by 1 microM diazepam. The currents showed outward rectification. Channels activated at depolarized 40 mV relative to the chloride reversal potential had low conductance (<40 pS) but the conductance increased with time, resulting in high-conductance channels (>40 pS). The average maximal-channel conductances for 2 and 100 microM THIP were 59 and 62 pS (-Vp = 40 mV), respectively, whereas in 2 microM THIP plus 1 microM diazepam, it was 71 pS. The results show that in hippocampal neurons THIP activates channels with characteristics similar to those of channels activated by low concentrations (0.5 5 microM ) of GABA. The increase in the inhibitory conductance with membrane depolarization permits gradation of the shunt pathway relative to the level of the excitatory input. PMID- 14550785 TI - Mechanisms of ectodermal organogenesis. AB - All ectodermal organs, e.g. hair, teeth, and many exocrine glands, originate from two adjacent tissue layers: the epithelium and the mesenchyme. Similar sequential and reciprocal interactions between the epithelium and mesenchyme regulate the early steps of development in all ectodermal organs. Generally, the mesenchyme provides the first instructive signal, which is followed by the formation of the epithelial placode, an early signaling center. The placode buds into or out of the mesenchyme, and subsequent proliferation, cell movements, and differentiation of the epithelium and mesenchyme contribute to morphogenesis. The molecular signals regulating organogenesis, such as molecules in the FGF, TGFbeta, Wnt, and hedgehog families, regulate the development of all ectodermal appendages repeatedly during advancing morphogenesis and differentiation. In addition, signaling by ectodysplasin, a recently identified member of the TNF family, and its receptor Edar is required for ectodermal organ development across vertebrate species. Here the current knowledge on the molecular regulation of the initiation, placode formation, and morphogenesis of ectodermal organs is discussed with emphasis on feathers, hair, and teeth. PMID- 14550786 TI - Cardiac T-box factor Tbx20 directly interacts with Nkx2-5, GATA4, and GATA5 in regulation of gene expression in the developing heart. AB - Tbx20 is a member of the T-box transcription factor family expressed in the forming hearts of vertebrate and invertebrate embryos. We report here analysis of Tbx20 expression during murine cardiac development and assessment of DNA-binding and transcriptional properties of Tbx20 isoforms. Tbx20 was expressed in myocardium and endocardium, including high levels in endocardial cushions. cDNAs generated by alternative splicing encode at least four Tbx20 isoforms, and Tbx20a uniquely carried strong transactivation and transrepression domains in its C terminus. Isoforms with an intact T-box bound specifically to DNA sites resembling the consensus brachyury half site, although with less avidity compared with the related factor, Tbx5. Tbx20 physically interacted with cardiac transcription factors Nkx2-5, GATA4, and GATA5, collaborating to synergistically activate cardiac gene expression. Among cardiac GATA factors, there was preferential synergy with GATA5, implicated in endocardial differentiation. In Xenopus embryos, enforced expression of Tbx20a, but not Tbx20b, led to induction of mesodermal and endodermal lineage markers as well as cell migration, indicating that the long Tbx20a isoform uniquely bears functional domains that can alter gene expression and developmental behaviour in an in vivo context. We propose that Tbx20 plays an integrated role in the ancient myogenic program of the heart, and has been additionally coopted during evolution of vertebrates for endocardial cushion development. PMID- 14550787 TI - Cortical and retinal defects caused by dosage-dependent reductions in VEGF-A paracrine signaling. AB - To determine the function of VEGF-A in nervous system development, we have utilized the Nestin promoter-driven Cre recombinase transgene, in conjunction with a conditional and hypomorphic VEGF-A allele, to lower VEGF-A activity in neural progenitor cells. Mice with intermediate levels of VEGF-A activity showed decreased blood vessel branching and density in the cortex and retina, resulting in a thinner retina and aberrant structural organization of the cortex. Severe reductions in VEGF-A led to decreases in vascularity and subsequent hypoxia, resulting in the specific degeneration of the cerebral cortex and neonatal lethality. Decreased neuronal proliferation and hypoxia was evident at E11.5, leading to increased neuronal apoptosis in the cortex by E15.5. In order to address whether the observed changes in the structural organization of the nervous system were due to a direct and autocrine role of VEGF-A on the neural population, we conditionally inactivated the main VEGF-A receptor, Flk1, specifically in neuronal lineages, by using the Nestin Cre transgene. The normality of these mice ruled out the possibility that VEGF-A/Flk1 signaling has a significant autocrine role in CNS development. VEGF-A dosage is therefore a critical parameter regulating the density of the vascular plexus in the developing CNS that is in turn a key determinant in the development and architectural organization of the nervous system. PMID- 14550788 TI - Insulin-like growth factor 1 is required for survival of transit-amplifying neuroblasts and differentiation of otic neurons. AB - Neurons that connect mechanosensory hair cell receptors to the central nervous system derive from the otic vesicle from where otic neuroblasts delaminate and form the cochleovestibular ganglion (CVG). Local signals interact to promote this process, which is autonomous and intrinsic to the otic vesicle. We have studied the expression and activity of insulin-like growth factor-1 (IGF-1) during the formation of the chick CVG, focusing attention on its role in neurogenesis. IGF-1 and its receptor (IGFR) were detected at the mRNA and protein levels in the otic epithelium and the CVG. The function of IGF-1 was explored in explants of otic vesicle by assessing the formation of the CVG in the presence of anti-IGF-1 antibodies or the receptor competitive antagonist JB1. Interference with IGF-1 activity inhibited CVG formation in growth factor-free media, revealing that endogenous IGF-1 activity is essential for ganglion generation. Analysis of cell proliferation cell death, and expression of the early neuronal antigens Tuj-1, Islet-1/2, and G4 indicated that IGF-1 was required for survival, proliferation, and differentiation of an actively expanding population of otic neuroblasts. IGF 1 blockade, however, did not affect NeuroD within the otic epithelium. Experiments carried out on isolated CVG showed that exogenous IGF-1 induced cell proliferation, neurite outgrowth, and G4 expression. These effects of IGF-1 were blocked by JB1. These findings suggest that IGF-1 is essential for neurogenesis by allowing the expansion of a transit-amplifying neuroblast population and its differentiation into postmitotic neurons. IGF-1 is one of the signals underlying autonomous development of the otic vesicle. PMID- 14550789 TI - Nlz belongs to a family of zinc-finger-containing repressors and controls segmental gene expression in the zebrafish hindbrain. AB - The zebrafish nlz gene has a rostral expression limit at the presumptive rhombomere (r) 3/r4 boundary during gastrula stages, and its expression progressively expands rostrally to encompass both r3 and r2 by segmentation stages, suggesting a role for nlz in hindbrain development. We find that Nlz is a nuclear protein that associates with the corepressor Groucho, suggesting that Nlz acts to repress transcription. Consistent with a role as a repressor, misexpression of nlz causes a loss of gene expression in the rostral hindbrain, likely due to ectopic nlz acting prematurely in this domain, and this repression is accompanied by a partial expansion in the expression domains of r4-specific genes. To interfere with endogenous nlz function, we generated a form of nlz that lacks the Groucho binding site and demonstrate that this construct has a dominant negative effect. We find that interfering with endogenous Nlz function promotes the expansion of r5 and, to a lesser extent, r3 gene expression into r4, leading to a reduction in the size of r4. We conclude that Nlz is a transcriptional repressor that controls segmental gene expression in the hindbrain. Lastly, we identify additional nlz-related genes, suggesting that Nlz belongs to a family of zinc-finger proteins. PMID- 14550790 TI - Pitx2c attenuation results in cardiac defects and abnormalities of intestinal orientation in developing Xenopus laevis. AB - The experimental manipulation of early embryologic events, resulting in the misexpression of the homeobox transcription factor pitx2, is associated with subsequent defects of laterality in a number of vertebrate systems. To clarify the role of one pitx2 isoform, pitx2c, in determining the left-right axis of amphibian embryos, we examined the heart and gut morphology of Xenopus laevis embryos after attenuating pitx2c mRNA levels using chemically modified antisense oligonucleotides. We demonstrate that the partial depletion of pitx2c mRNA in these embryos results in alteration of both cardiac morphology and intestinal coiling. The most common cardiac abnormality seen was a failure of rightward migration of the outflow tract, while the most common intestinal laterality phenotype seen was a full reversal in the direction of coiling, each present in 23% of embryos injected with the pitx2c antisense oligonucleotide. An abnormality in either the heart or gut further predisposed to a malformation in the other. In addition, a number of other cardiac anomalies were observed after pitx2c mRNA attenuation, including abnormalities of atrial septation, extracellular matrix restriction, relative atrial-ventricular chamber positioning, and restriction of ventricular development. Many of these findings correlate with cardiac defects previously reported in pitx2 null and hypomorphic mice, but can now be assigned specifically to attenuation of the pitx2c isoform in Xenopus. PMID- 14550791 TI - Isolation of Rho GTPase effector pathways during axon development. AB - The Rho GTPases Rac1 and Cdc42 have been implicated in the regulation of axon outgrowth and guidance. However, the downstream effector pathways through which these GTPases exert their effects on axon development are not well characterized. Here, we report that axon outgrowth defects within specific subsets of motoneurons expressing constitutively active Drosophila Rac1 largely persist even with the addition of an effector-loop mutation to Rac1 that disrupts its ability to bind to p21-activated kinase (Pak) and other Cdc42/Rac1 interactive-binding (CRIB)-motif effector proteins. While hyperactivation of Pak itself does not lead to axon outgrowth defects as when Rac1 is constitutively activated, live analysis reveals that it can alter filopodial activity within specific subsets of neurons similar to constitutive activation of Cdc42. Moreover, we show that the axon guidance defects induced by constitutive activation of Cdc42 persist even in the absence of Pak activity. Our results suggest that (1) Rac1 controls axon outgrowth through downstream effector pathways distinct from Pak, (2) Cdc42 controls axon guidance through both Pak and other CRIB effectors, and (3) Pak's primary contribution to in vivo axon development is to regulate filopodial dynamics that influence growth cone guidance. PMID- 14550792 TI - The CBP coactivator functions both upstream and downstream of Dpp/Screw signaling in the early Drosophila embryo. AB - The CBP histone acetyltransferase plays important roles in development and disease by acting as a transcriptional coregulator. A small reduction in the amount of Drosophila CBP (dCBP) leads to a specific loss of signaling by the TGF beta molecules Dpp and Screw in the early embryo. We show that the expression of Screw itself, and that of two regulators of Dpp/Screw activity, Twisted gastrulation and the Tolloid protease, is compromised in dCBP mutant embryos. This prevents Dpp/Screw from initiating a signal transduction event in the receiving cell. Smad proteins, the intracellular transducers of the signal, fail to become activated by phosphorylation in dCBP mutants, leading to diminished Dpp/Screw-target gene expression. At a slightly later stage of development, Dpp/Screw-signaling recovers in dCBP mutants, but without a restoration of Dpp/Screw-target gene expression. In this situation, dCBP acts downstream of Smad protein phosphorylation, presumably via direct interactions with the Drosophila Smad protein Mad. It appears that a major function of dCBP in the embryo is to regulate upstream components of the Dpp/Screw pathway by Smad-independent mechanisms, as well as acting as a Smad coactivator on downstream target genes. These results highlight the exceptional sensitivity of components in the TGF-beta signaling pathway to a decline in CBP concentration. PMID- 14550794 TI - Transgenic zebrafish with fluorescent germ cell: a useful tool to visualize germ cell proliferation and juvenile hermaphroditism in vivo. AB - Juvenile zebrafish are hermaphroditic; undifferentiated gonads first develop into ovary-like tissues, which then either become ovaries and produce oocytes (female) or degenerate and develop into testes (male). In order to fully capture the dynamic processes of germ cells' proliferation and juvenile hermaphroditism in zebrafish, we established transgenic lines TG(beta-actin:EGFP), harboring an enhanced green fluorescent protein (EGFP) gene driven by a medaka beta-actin promoter. In TG(beta-actin:EGFP), proliferating germ cells and female gonads strongly expressed EGFP, but fluorescence was only dimly detected in male gonads. Based on the fluorescent (+) or nonfluorescent (-) appearance of germ cells seen in living animals, three distinct groups were evident among TG(beta-actin:EGFP). Transgenics in ++ group (44%) were females, had fluorescent germ cells as juveniles, and female gonads continuously fluoresced throughout sexual maturation. Transgenics in +- (23%) and -- (33%) groups were males. Fluorescent germ cells were transiently detected in +- transgenics from 14 to 34 days postfertilization (dpf), but were not detected in -- transgenics throughout their life span. Histological analyses showed that 26-dpf-old transgenics in ++, +-, and -- groups all developed ovary-like tissues: Germ cells in -- group juveniles arrested at the gonocyte stage and accumulated low quantities of EGFP, while those in ++ group juveniles highly proliferated into diplotene to perinucleolar stages and accumulated high quantities of EGFP. In +- group juveniles, degenerating oocytes, gonocytes, and spermatogonia were coexistent in transiently fluorescent gonads. Therefore, the fluorescent appearance of gonads in this study was synchronous with the differentiation of ovary-like tissues. Thus, TG(beta actin:EGFP) can be used to visualize germ cells' proliferation and juvenile hermaphroditism in living zebrafish for the first time. PMID- 14550793 TI - Sexual differentiation of germ cells in XX mouse gonads occurs in an anterior-to posterior wave. AB - Differentiation of mouse embryonic germ cells as male or female is dependent on the somatic environment of the gonad rather than the sex chromosome constitution of the germ cell. However, little is known about the initiation of germ cell sexual differentiation. Here, we traced the initiation of germ cell sexual differentiation in XX gonads using the Stra8 gene, which we demonstrate is an early molecular marker of female germ cell development. Stra8 is upregulated in embryonic germ cells of XX gonads prior to meiotic entry and is not expressed in male embryonic germ cells. A developmental time course of Stra8 expression in germ cells of XX gonads has revealed an anterior-to-posterior wave of differentiation that lasts approximately 4 days, from embryonic days 12.5 to 16.5. Consistent with these results, we find that embryonic ovarian germ cells upregulate the meiotic gene Dmc1 and downregulate the Oct4 transcription factor in an anterior-to-posterior wave. In complementary experiments, we find that embryonic XX gonads upregulate certain gene markers of somatic female differentiation in an anterior-to-posterior pattern, while others display a center-to-pole pattern of regulation. Thus, sexual differentiation and meiotic entry of germ cells in embryonic XX gonads progress in an anterior-to-posterior pattern that may reflect local environmental cues that are present in the embryonic XX gonad. PMID- 14550795 TI - Membrane segregation and downregulation of raft markers during sarcolemmal differentiation in skeletal muscle cells. AB - Muscle contraction implies flexibility in combination with force resistance and requires a high degree of sarcolemmal organization. Smooth muscle cells differentiate largely from mesenchymal precursor cells and gradually assume a highly periodic sarcolemmal organization. Skeletal muscle undergoes an even more striking differentiation programme, leading to cell fusion and alignment into myofibrils. The lipid bilayer of each cell type is further segregated into raft and non-raft microdomains of distinct lipid composition. Considering the extent of developmental rearrangement in skeletal muscle, we investigated sarcolemmal microdomain organization in skeletal and smooth muscle cells. The rafts in both muscle types are characterized by marker proteins belonging to the annexin family which localize to the inner membrane leaflet, as well as glycosyl-phosphatidyl inositol (GPI)-anchored enzymes attached to the outer leaflet. We demonstrate that the profound structural rearrangements that occur during skeletal muscle maturation coincide with a striking decrease in membrane lipid segregation, downregulation of annexins 2 and 6, and a significant decrease in raft-associated 5'-nucleotidase activity. The relative paucity of lipid rafts in mature skeletal in contrast to smooth muscle suggests that the organization of sarcolemmal microdomains contributes to the muscle-specific differences in stimulatory responses and contractile properties. PMID- 14550796 TI - Ventral dominance governs sequential patterns of gene expression across the dorsal-ventral axis of the neuroectoderm in the Drosophila embryo. AB - A nuclear concentration gradient of the maternal transcription factor Dorsal establishes three tissues across the dorsal-ventral axis of precellular Drosophila embryos: mesoderm, neuroectoderm, and dorsal ectoderm. Subsequent interactions among Dorsal target genes subdivide the mesoderm and dorsal ectoderm. Here we investigate the subdivision of the neuroectoderm by three conserved homeobox genes, ventral nervous system defective (vnd), intermediate neuroblasts defective (ind), and muscle segment homeobox (msh). These genes divide the ventral nerve cord into three columns along the dorsal-ventral axis. Sequential patterns of vnd, ind, and msh expression are established prior to gastrulation and evidence is presented that these genes respond to distinct thresholds of the Dorsal gradient. Maintenance of these patterns depends on cross regulatory interactions, whereby genes expressed in ventral regions repress those expressed in more dorsal regions. This "ventral dominance" includes regulatory genes that are expressed in the mesectoderm and mesoderm. At least some of these regulatory interactions are direct. For example, the misexpression of vnd in transgenic embryos represses ind and msh, and the addition of Vnd binding sites to a heterologous enhancer is sufficient to mediate repression. The N-terminal domain of Vnd contains a putative eh1 repression domain that binds Groucho in vitro. Mutations in this domain diminish Groucho binding and also attenuate repression in vivo. We discuss the significance of ventral dominance with respect to the patterning of the vertebrate neural tube, and compare it with the previously observed phenomenon of posterior prevalence, which governs sequential patterns of Hox gene expression across the anterior-posterior axis of metazoan embryos. PMID- 14550797 TI - Polyhomeotic stably associates with molecular chaperones Hsc4 and Droj2 in Drosophila Kc1 cells. AB - Polycomb group (PcG) proteins silence target loci in Drosophila. Although the mechanism of PcG-mediated silencing remains unknown, there is considerable evidence that PcG proteins act via multiple complexes. We have epitope-tagged Polyhomeotic Proximal, PHP, the major isoform of the proximal product of the polyhomeotic locus, at both termini (F-PHP-HA) and generated a stable Kc1 cell line in order to isolate F-PHP-HA-associated proteins. Using either column chromatography followed by immunoaffinity precipitation or a double immunoaffinity precipitation procedure, we observed multiple proteins that stably associate with F-PHP-HA. Sequencing the five major bands identified PHP-170 and PHP-140 isoforms, Polycomb, Heat shock cognate 4 (Hsc4), and a novel Drosophila J class chaperone we term Droj2. Mutations in both chaperone genes enhance homeotic transformations in PcG genes, suggesting that they have a role in silencing. We show by Western blotting that minor components of F-PHP-HA-associated proteins include TBP, TAF(II)42, TAF(II)85, and p55. However, unlike in PRC1, Psc, TAF(II)62, Modulo, dMI-2, or Rpd3/HDAC1 do not associate with F-PHP-HA. We discuss the role of chaperones and F-PHP-HA-associated proteins in PcG-mediated silencing and the evidence for different complexes containing Polyhomeotic in vivo. PMID- 14550798 TI - Introducing the special issue on allogeneic hematopoietic stem cell transplantation. PMID- 14550799 TI - Aims of conditioning. AB - Nuclear warfare research and treatment of radiation accident victims uncovered the potential of hemopoietic stem cell transplants. Prior to transplantation of hemopoietic stem cells patients receive "conditioning" agents: high-dose total body irradiation and/or high-dose chemotherapy. High-dose conditioning causes at least 20% procedure-related mortality. Recent efforts to reduce procedure-related mortality by the use of low-dose conditioning included low-dose total-body irradiation, immunosuppressive agents, and the replacement of high-dose chemotherapy by donor lymphocytes for graft-vs-tumor effects. Procedure-related mortality remains high (10-30%). Tumor recurrence at 1 year is over 50%. In this review, the aims of conditioning (creation of space, prevention of hemopoietic stem cell rejection, eradication of immune memory, and eradication of tumor cells) are reexamined in those patient and animal studies that explore quantitative and mechanistic conditioning issues. Translational experimental animal models provide the best opportunities for the development of less toxic conditioning agents for human patients and require an analysis of the consequences of the effects of new conditioning agents on host-vs-graft as well as graft-vs-host reactions. Total-body irradiation or other forms of radiation create space, prevent rejection of histocompatible stem cells, and can eliminate immune memory to autoimmune antigens at modest, nontoxic doses. The transplantation of histoincompatible stem cells and the eradication of large loads of tumor cells remain problematic. The therapeutic index of allogeneic stem cell transplants will increase if new conditioning agents are targeted only to those host tissues that need conditioning: hemopoietic system, immune system, and tumor masses. Radiolabeled immunoglobulins are among the most promising new, low toxicity conditioning agents. PMID- 14550801 TI - Minimal GVHD following in-vitro T cell-depleted allogeneic stem cell transplantation with reduced-intensity conditioning allowing subsequent infusions of donor lymphocytes in patients with hematological malignancies and solid tumors. AB - OBJECTIVE: Allogeneic stem cell transplantation (alloSCT) following reduced intensity conditioning offers a relatively nontoxic regimen while preserving rapid and sustained engraftment. Acute and chronic graft-vs-host disease (GVHD) is, however, a significant cause of severe morbidity. To reduce the incidence of GVHD, we treated a group of high-risk patients with a reduced-intensity conditioning regimen followed by in vitro T-cell-depleted alloSCT using Campath 1 H incubation. PATIENTS AND METHODS: Eighteen patients were treated with fludarabine (6 x 30 mg/m(2)), busulphan (2 x 3.2 mg/kg), and ATG (4 x 10 mg/kg) followed by the infusion of high-dose T-cell-depleted peripheral stem cells from sibling donors. No posttransplant GVHD prophylaxis was administered. At 6 months after alloSCT, low-dose donor lymphocyte infusion (DLI) was administered. RESULTS: All patients had sustained engraftment of donor cells with a median of 95% donor cells at 3 months after alloSCT. Minimal acute and no chronic GVHD was observed after alloSCT. A high incidence of cytomegalovirus (CMV) reactivation but no CMV disease was observed. Eleven patients received DLI at a median of 6.5 months after alloSCT. Acute GVHD grade II-III developed in 6 patients. All patients showed improvement of donor chimerism after DLI. With a median follow-up of 211 days, 11 patients are alive. Particular in patients with chronic lymphocytic leukemia and acute myeloid leukemia, a significant graft-vs-tumor effect was observed. CONCLUSIONS: In vitro T-cell-depleted alloSCT following reduced-intensity conditioning leads to durable donor engraftment without GVHD. The high levels of donor chimerism allow the subsequent use of cellular immunotherapy to treat residual disease. PMID- 14550800 TI - A randomized multicenter comparison of CD34(+)-selected progenitor cells from blood vs from bone marrow in recipients of HLA-identical allogeneic transplants for hematological malignancies. AB - OBJECTIVE: Peripheral blood progenitor cells (PBPC) have been established as an alternative source of hematopoietic stem cells for allogeneic transplantation, but an increased incidence of both acute and chronic graft-vs-host disease (GVHD) has become apparent. We performed a prospective randomized trial comparing bone marrow transplantation (BMT) vs PBPC transplantation (PBPCT) using CD34(+) selection for T-cell depletion (TCD) in both study arms. PATIENTS AND METHODS: Between January 1996 and October 2000, 120 patients with a diagnosis of acute leukemia, myelodysplasia, multiple myeloma, or lymphoma were randomized to receive either filgrastim-mobilized PBPC or BM from HLA-identical sibling donors after standard high-dose chemoradiotherapy. Patient characteristics did not differ between study arms. RESULTS: Recipients of PBPC received more CD3(+) T cells (median: 3.0 vs 2.0 x 10(5)/kg, p<0.0001) and more CD34(+) cells (median: 3.6 vs 0.9 x 10(6)/kg, p<0.0001). Neutrophil and platelet recoveries occurred significantly faster after PBPCT. The cumulative incidence of acute GVHD grades II-IV was 37% after BMT vs 52% after PBPCT and was most significantly (p=0.007) affected by the number of CD3(+) T cells in the graft. Acute GVHD appeared strongly associated with increased treatment-related mortality (TRM) in a time dependent analysis. Higher numbers of CD34(+) cells were associated with less TRM. With a median follow-up of 37 months (range: 12-75), overall survival at 4 years from transplantation was 60% after BMT and 34% for recipients of PBPCT (p=0.04), which difference was largely due to increased GVHD and TRM in PBPC recipients receiving T-cell dosages greater than 2 x 10(5)/kg. CONCLUSION: Outcome following T cell-depleted PBPCT critically depends on the number of CD3(+) T cells, whereby high T-cell numbers may blunt a favorable effect of higher CD34(+) cell numbers. PMID- 14550802 TI - Bone marrow as stem cell source for allogeneic HLA-identical sibling transplantation following reduced-intensity preparative regimen. AB - OBJECTIVE: Reduced-intensity conditioning regimens (RIC) and peripheral blood stem cells (PBSC) are increasingly used for allogeneic stem cell transplantation (allo-BMT). RIC has been shown to allow engraftment with minimal early transplant related mortality (TRM). However, in the context of RIC, the use of bone marrow (BM) as stem cell source is still little evaluated. PATIENTS AND METHODS: In this report, we analyzed the outcome of 32 high-risk patients with hematological malignancies who received an HLA-identical sibling allo-BMT after RIC including fludarabine, busulfan, and anti-thymocyte globulin (ATG). RESULTS: Sustained neutrophil and platelet recovery occurred at a median of 13 days (range, 10-19) and 17 days (range, 0-45) respectively. Early and durable full donor chimerism could be established as soon as the first month after allo-BMT. Also, a sustained and early CD8(+) T-cell recovery was observed, but the CD4(+) T-cell compartment remained profoundly low. The cumulative incidences of grade II-IV acute GVHD and chronic GVHD were 26% (95% CI, 11-41%) and 31% (95% CI, 15-47%) respectively. The overall cumulative incidence of TRM was 28% (95% CI, 12-44%) occurring mainly in patients aged over 50. In this setting, GVHD showed a protective effect on disease progression or relapse with better progression-free survival for patients with GVHD as compared to patients without GVHD (p=0.03). CONCLUSIONS: Collectively, these results confirm that the use of BM grafts for RIC is feasible with durable donor engraftment and no detrimental GVHD. PMID- 14550803 TI - Nonexpanded primary lung and bone marrow-derived mesenchymal cells promote the engraftment of umbilical cord blood-derived CD34(+) cells in NOD/SCID mice. AB - OBJECTIVE: Previously, we have found that human culture-expanded fetal lung derived mesenchymal stem cells (MSC) promote the engraftment of umbilical cord blood (UCB)-derived CD34((+)) cells. The high frequency of MSC in fetal lung allowed us to study whether this represented a biological feature of these cells or a property that was acquired during expansion in culture. MATERIALS AND METHODS: Irradiated NOD/SCID mice (n=80) were transplanted with 0.1x10(6) UCB CD34(+) cells in the presence or absence of 10(6) primary nonexpanded or culture expanded fetal lung, liver, or BM CD45(-) cells, or with nonexpanded fetal lung liver or BM CD45(-) cells only. RESULTS: In comparison with transplantation of UCB CD34(+) cells only, cotransplantation of UCB CD34(+) cells and primary fetal lung or BM CD45(-) cells resulted in a significantly higher level of engraftment (% hCD45(+) cells) in BM, PB, and spleen. In addition, primary mesenchymal cells derived from adult BM had a similar promoting effect. The engraftment-enhancing effect was similar to that of culture-expanded fetal lung and BM MSC. Primary mesenchymal cells, but not culture-expanded MSC, were detected in recipient mice, suggesting that the primary cells were able to home and that this capacity was lost after expansion. CONCLUSIONS: These results show that primary mesenchymal cells from fetal lung and BM promote the engraftment of UCB-derived CD34(+) cells to a similar degree as culture-expanded MSC, indicating that it reflects a biological property of primary MSC that is preserved during expansion in culture. PMID- 14550804 TI - HLA expression and immunologic properties of differentiated and undifferentiated mesenchymal stem cells. AB - OBJECTIVE: Mesenchymal stem cells (MSC) do not elicit alloreactive lymphocyte responses due to immune modulations. We investigated the immunologic properties of MSC after differentiation along three lineages: bone, cartilage, and adipose. METHODS AND RESULTS: Flow cytometry showed that undifferentiated MSC express HLA class I but not class II, although HLA class II was present intracellularly as detected by Western blot. Addition of interferon gamma (IFN-gamma) for 48 hours induced greater than 90% of cells to express HLA class II. No lymphocyte response was induced by allogeneic irradiated MSC as stimulators. Results were similar using MSC pretreated with IFN-gamma. After growth of cells in medium to induce differentiation to bone, cartilage, or adipose for 6 or 12 days, the expression of HLA class I increased but no class II was detected on the cell surface. The ability to upregulate HLA class II on the cell surface after exposure to IFN gamma for 48 hours was clearly diminished after the cells had been cultured in differentiation medium for 6 or 12 days, with only 10% of cells expressing HLA class II. Using MSC grown in osteogenic, chondrogenic, or adipogenic medium as stimulator cells, no lymphocyte alloreactivity was seen, even if differentiated MSC had been pretreated with IFN-gamma. MSC inhibit mixed lymphocyte cultures, particularly after osteogenic differentiation. This suppression was further enhanced by IFN-gamma. CONCLUSIONS: Undifferentiated and differentiated MSC do not elicit alloreactive lymphocyte proliferative responses and modulate immune responses. The findings support that MSC can be transplantable between HLA incompatible individuals. PMID- 14550806 TI - Herpes simplex thymidine kinase gene-transduced donor lymphocyte infusions. AB - OBJECTIVE: Donor lymphocytes mediate both a beneficial graft-vs-leukemia/lymphoma (GVL) effect as well as graft-vs-host disease (GVHD), the most dreaded complication of allogeneic hematopoietic stem cell transplantation (HSCT). Transduction of donor lymphocytes with a herpes simplex thymidine kinase (HSVtk) gene prior to infusion confers lethal sensitivity to the anti-herpes drug, ganciclovir (GCV). HSVtk-transduced donor lymphocyte infusions (DLI) have already been used and significant problems have limited the clinical experience to very few patients. To this end, we also report on a study of whether HSVtk-DLI induces GVHD/GVL and if infusion of GCV allows abrogation of GVHD by selective killing of donor lymphocytes. MATERIALS AND METHODS: Nine patients with relapsed hematologic malignancies after allogeneic hematopoietic stem cell transplantation (HSCT) were infused with HSVtk gene-modified donor lymphocytes. In brief, transgeneic lymphocytes were prepared by 3 days of activation, 1 day of transduction, 6 days of selection with G418, and 2 to 4 weeks of expansion. RESULTS: From 5.0 to 199 x 10(6) CD3(+) DLI were infused. There were no toxicities and no correlation between CD3(+) cell dose and either GVHD or GVL was observed. Only one patient who had cutaneous T-cell lymphoma (CTCL) developed GVHD and that same patient is the only patient to have an anti-tumor response. The patient was infused with 23 x 10(6) CD4(+) and 9.7 x 10(6) CD8(+) HSVtk DLI. Following discontinuation of immune suppression and infusion of GCV, GVHD promptly resolved. Although the CTCL relapsed, it has been easily controlled with intermittent topical therapy. One patient with acute myelogenous leukemia (AML) had a remission inversion of undetermined significance. Two patients with AML, one patient with lymphoma, and four patients with chronic myelogenous leukemia (CML) did not respond. CONCLUSION: HSVtk-DLI may provide an anti-tumor effect in vivo and may induce GVHD that is abrogated by GCV treatment. While technical aspects to improve response need to be perfected, HSVtk-DLI infusion to induce a transient GVL/GVHD may become an effective future therapy to minimize complications of allogeneic HSCT. PMID- 14550805 TI - Role of CXCR3-induced donor T-cell migration in acute GVHD. AB - OBJECTIVE: The chemokine receptor CXCR3 has an important role in the migration of effector T cells. To investigate the role of CXCR3 on donor cells in acute graft vs host disease (GVHD) we used a well-defined experimental bone marrow transplantation (BMT) model where acute GVHD is mediated by donor CD8(+) T cells against minor histocompatibility antigens. METHODS; Lethally irradiated C3H.SW recipients were transplanted from either wild-type B6 or CXCR3(-/-) B6 donors. Donor T-cell expansion was analyzed in the spleen and small intestine of recipients by FACS. Donor T-cell function was analyzed by cytokine secretion. The severity of acute GVHD was assessed by histopathological analysis of intestine and liver, GVHD clinical scores, and survival after BMT. RESULTS: Significantly higher numbers of donor CD8(+) CXCR3(-/-) T cells were found in the spleen on days +7 and +14 compared to donor wild-type T cells. By contrast, the number of CD8(+) T cells in the small bowel of BMT recipients from CXCR3(-/-) donors was sevenfold lower than from wild-type donors. Systemic concentrations of INF-gamma and TNF-alpha were equivalent between groups. Animals that received CXCR3(-/-) donor T cells demonstrated diminished GI tract and liver damage and showed improved survival after BMT compared to recipients of wild-type donor cells (43% vs 0%, p<0.001). CONCLUSION: The migration of donor CD8(+) T cells to GVHD target organs such as the intestine depends on the expression of CXCR3 and contributes significantly to GVHD damage and overall mortality. PMID- 14550807 TI - NK cell recovery, chimerism, function, and recognition in recipients of haploidentical hematopoietic cell transplantation following nonmyeloablative conditioning using a humanized anti-CD2 mAb, Medi-507. AB - OBJECTIVE: Natural killer (NK) cells kill allogeneic cells that lack a class I MHC ligand for clonally distributed killer inhibitory receptors (KIR). Following HLA-mismatched hematopoietic cell transplantation (HCT), donor NK cells might mediate graft-vs-host (GVH) reactions that promote donor chimerism and mediate anti-tumor effects. Additionally, recipient NK cells might mediate donor marrow rejection. We have developed a nonmyeloablative approach to haploidentical HCT involving recipient treatment with a T cell-depleting mAb, Medi-507, that can achieve donor engraftment and mixed hematopoietic chimerism without graft-vs-host disease (GVHD). Donor lymphocyte infusions (DLI) are later administered in an effort to achieve graft-vs-leukemia/lymphoma (GVL) effects without GVHD. It is unknown whether NK cell "tolerance" develops in human mixed chimeras. METHODS: We have addressed these issues in 12 patients receiving Medi-507-based nonmyeloablative haploidentical HCT. RESULTS: NK cells recovered relatively early, despite the presence of circulating anti-CD2 mAb, but the majority of initially recovering cells lacked CD2 expression. These NK cells showed a reduced capacity, compared to those from normal donors, to kill class I-deficient targets. No association was detected between KIR mismatches in the host-vs-graft (HVG) or GVH direction and graft or tumor outcomes in this small series. NK cell chimerism did not correlate with chimerism in other lineages in mixed chimeras. NK cell tolerance to the host was not observed in a patient with full donor chimerism. One patient developed NK cell reactivity against donor-derived lymphoblast targets after loss of chimerism, despite the absence of an HVG KIR mismatch. CONCLUSION: Our results do not show an impact of NK cells on the outcome of nonmyeloablative, even T cell-depleted, HCT across haplotype barriers using an anti-CD2 mAb. Our data also raise questions about the applicability of observations made with NK cell clones to the bulk NK cell repertoire in humans. PMID- 14550808 TI - Molecular assessment of erythroid lineage chimerism following nonmyeloablative allogeneic stem cell transplantation. AB - OBJECTIVE: Nonmyeloablative conditioning regimens for allogeneic stem cell transplantation are now commonly used in the treatment of patients with hematologic malignancies. Since this treatment often results in the establishment of mixed hematopoietic chimerism, this approach may also prove to be useful in the treatment of nonmalignant disorders, such as sickle cell disease and thalassemia major. To apply this approach to these diseases, it will be necessary to determine the levels of donor erythropoiesis required to correct hemolysis and ameliorate disease symptoms. Current methods for measuring hematopoietic chimerism are based on DNA polymorphisms that distinguish recipient from donor. These methods accurately measure donor leukocyte engraftment but do not quantify the relative contributions of recipient and donor erythropoiesis following transplant. METHODS: To specifically measure erythroid-lineage chimerism, we used pyrosequencing of the sickle cell mutation to quantify the relative levels of normal and sickle beta-globin mRNA in patient samples. Results of beta-globin RNA chimerism were compared to assessment of beta-globin DNA chimerism as well as analysis of short tandem repeat (STR) polymorphisms, cytogenetics, and hemoglobin electrophoresis. RESULTS: Donor engraftment was measured in two adult patients following nonmyeloablative stem cell transplant for sickle cell disease. In Patient 1, 25 to 30% of peripheral leukocytes were donor derived after day 41. In contrast, more than 55% of peripheral blood beta-globin mRNA was of donor origin, and these results correlated with posttransplant clinical improvement. Patient 2 achieved 40 to 50% donor leukocyte engraftment from day 33 onward. This was associated with 70 to 100% peripheral blood donor beta-globin mRNA. CONCLUSIONS: These studies demonstrate that relatively low levels of donor leukocyte engraftment can be associated with higher levels of donor erythropoiesis and with significant clinical improvement. Pyrosequencing of lineage-specific mRNA directly measures functional reconstitution of donor cells and provides valuable information that can affect clinical decisions in patients with nonmalignant diseases following allogeneic transplant. PMID- 14550809 TI - Low-dose total-body irradiation and fludarabine followed by hematopoietic cell transplantation from HLA-identical sibling donors do not induce complete T-cell donor engraftment in most patients with progressive hematologic diseases. AB - OBJECTIVE: The aim of this study was to determine whether nonmyeloablative transplants (NMT) result in complete and sustained donor engraftment in patients with progressive hematologic diseases compared to patients with stable disease or who are in remission. MATERIALS AND METHODS: We prospectively monitored the kinetics of engrafting of T cells and myeloid cells in 10 consecutive adult patients with hematologic diseases submitted to NMT from an HLA-identical sibling donor. Patients were considered ineligible for conventional allogeneic transplantation because of age, concomitant diseases, or previous autologous transplant. Conditioning regimen and graft-vs-host disease posttransplant prophylaxis consisted of 2-Gy total-body irradiation plus fludarabine 30 mg/m(2)/day for 3 days, and cyclosporin and mycophenolate mofetil, respectively. RESULTS: One patient died in remission, and eight relapsed or progressed at a median of 68 days (15-335). On day +56, only 1 (11%) of 9 patients analyzed had achieved T-cell complete donor chimerism (CC), whereas 6 (67%) had achieved myeloid CC (p=0.05). Median time for T-cell CC to occur was 110 days (56-150) compared with 42 days (28-100) to achieve myeloid CC (p=0.002). The only parameter associated with T-cell CC was the status of the disease at the time of transplantation. Thus, 5 (100%) of 5 patients with stable disease or who were in remission before the transplant achieved T-cell CC compared with only 1 (20%) of 5 patients with progressive disease (p=0.05). CONCLUSION: Conditioning regimen based on fludarabine and 2-Gy total-body irradiation allows cell immunotherapy for old and medically infirm patients, but its antitumoral effect in patients with progressive hematologic disease is limited. PMID- 14550810 TI - Immunologic recovery after hematopoietic cell transplantation with nonmyeloablative conditioning. AB - OBJECTIVE: We studied immune reconstitution in 51 recipients of HLA-identical hematopoietic cellular transplant (HCT) after nonmyeloablative conditioning compared to a reference group of 67 recipients after myeloablative conditioning. METHODS: Nonmyeloablative conditioning consisted of 2 Gy total-body irradiation+/ fludarabine and postgrafting cyclosporine and mycophenolate mofetil. All patients received G-CSF-mobilized peripheral blood mononuclear cells. Patients were followed with serial assessments of lymphocyte subset counts, antibody levels, virus-induced lymphoproliferation, and limiting-dilution assays for cytomegalovirus (CMV) T helper (T(H)) cells. Rates of infections over the first year after transplant were calculated. RESULTS: During the first 180 days, absolute lymphocyte subset counts were similar (except higher total and memory B cell counts on day 80 in nonmyeloablative patients). At 1 year, however, total and naive CD4 counts, and naive CD8 counts, were higher in myeloablative patients. The levels of antibodies were similar at all time points and after vaccinations. The function of CD4 cells assessed by virus-induced lymphoproliferation was similar. However, the absolute counts of CMV T(H) cells were higher at days 30 and 90 (p=0.002 and p=0.0003, respectively) after nonmyeloablative conditioning. The rates of definite infections were lower for nonmyeloablative patients during the first 90 days, but were higher later. The higher number of CMV-specific T cells days 30 and 90 after nonmyeloablative HCT coincided with a lower rate of CMV infections during that time. CONCLUSION: The immunity of nonmyeloablative HCT recipients appears better than the immunity of conventional HCT recipients early, but not late, after HCT. PMID- 14550811 TI - Growth hormone accelerates immune recovery following allogeneic T-cell-depleted bone marrow transplantation in mice. AB - OBJECTIVE: To test in a murine model whether recombinant human growth hormone can promote immune recovery after allogeneic T-cell-depleted bone marrow transplantation. MATERIALS AND METHODS: Lethally irradiated (8.5 Gy) BALB/c mice (H2(d)) were transplanted with 5 x 10(6) T cell-depleted bone marrow cells from C57BL/6 mice (H2(b)). Recipient mice were injected intraperitoneally with recombinant human growth hormone (20 microg/dose/day) or saline for the first 4 weeks after transplantation. These animals were followed for phenotypic and functional immune recovery. RESULTS: Administration of human recombinant growth hormone improved the CD4(+) T-cell counts in peripheral blood on day +14 (44+/-14 vs 33+/-7/microL blood, p<0.05) and day +21 (281+/-109 vs 187+/-76/microL blood, p<0.01) compared with the saline control. These differences were no longer significant by day +28 despite continued growth hormone administration. Similar effects were also observed on CD8(+) T cells and B220(+) B cells. The improvements in peripheral T-cell counts were at least partially as a result of enhanced thymopoiesis because there was an increase in total thymocytes after treatment with growth hormone. T-cell-depleted bone marrow recipients treated with growth hormone rejected the third-party grafts faster than those treated with saline control (median survival time: 20 days vs 26 days, p<0.05). CONCLUSIONS: These data demonstrated that recombinant human growth hormone can accelerate phenotypic and functional immune reconstitution following allogeneic T cell-depleted bone marrow transplantation in mice. PMID- 14550812 TI - Allogeneic graft CD34(+) cell dose correlates with dendritic cell dose and clinical outcome, but not with dendritic cell reconstitution after transplant. AB - OBJECTIVE: This study examined whether the CD34(+) cell dose in allografts correlates with the dose of myeloid dendritic cells (mDC) and plasmacytoid DC (pDC), and with DC reconstitution and clinical outcome after a myeloablative HLA matched transplant. PATIENTS AND METHODS: Fifty-three patients were included in this study: 37 who had undergone a granulocyte colony-stimulating factor mobilized peripheral blood stem cells (PBSC) transplant from related donors and 16 who had undergone a marrow transplant from unrelated donors. The number of CD34(+) cells, lin(-)HLA-DR(+)CD11c(+) mDC, lin(-)HLA-DR(+)CD123(+) pDC, CD14(+) monocytes, and CD3(+)CD4(+), CD3(+)CD8(+), CD56(+), and CD19(+) lymphocytes was compared in the graft, as well as in the peripheral blood after transplant, in patients receiving more than versus less than or equal to the median number of CD34(+) cells in PBSC (5.78 x 10(6)/kg) or in marrow (2.8 x 10(6)/kg). RESULTS: A higher CD34(+) cell dose was associated with larger numbers of mDC in PBSC (p=0.01) and pDC in marrow grafts (p=0.004). However, neither mDC nor pDC recovery after transplant correlated with the number of CD34(+) cells infused. Finally, higher doses of CD34(+) cells appeared to negatively affect (p=0.02) the overall survival in PBSC transplantation and were associated with a trend for higher acute graft-vs-host disease in PBSC and lower acute graft-vs-host disease in marrow transplant. CONCLUSIONS: CD34(+) cell dose correlates with the dose of different DC subsets in PBSC and marrow grafts, but it does not affect DC reconstitution after transplant. Higher doses of CD34(+) cells in PBSC, but not in marrow, seem to adversely affect survival after transplant. PMID- 14550813 TI - A CTL epitope from human cytomegalovirus IE1 defined by combining prediction of HLA binding and proteasomal processing is the target of dominant immune responses in patients after allogeneic stem cell transplantation. AB - OBJECTIVE AND METHODS: In an attempt to define HCMV IE1-derived, HLA-A(*)0201 restricted epitopes, an advanced computer-based epitope prediction combining HLA binding and proteasomal cleavages in silico was performed. RESULTS: This prediction algorithm clearly confirmed VLEETSVML to be the most likely CTL epitope. By tetramer staining, HCMV pp65 NLVPMVATV-specific CD8(+) T cells were detectable in 18/24 HCMV seropositive HLA-A(*)0201-expressing individuals (median frequency 0.58%; range 0.1%-4.7%), and IE1 VLEETSVML-specific CD8(+) T cells in 5/24 (median frequency 2.1%; range 0.1%-4.3%), respectively (p<0.01). Also in recipients of an allogeneic SCT, VLEETSVML- and NLVPMVATV-specific CD8(+) T cells were detectable in comparable frequencies, but again the number of patients with detectable pp65-specific CD8(+) T cells was higher (p=0.014). In 4/15 individuals, all demonstrating IE1 VLEETSVML-specific CD8(+) T cells prior to peptide stimulation, VLEETSVML-specific T cell lines (purity of 42.6%-98.6% of all CD3(+)/CD8(+) T cells) were successfully generated after 2-4 weeks of culture using the IFN-gamma secretion assay. CONCLUSION: In conclusion, this novel prediction strategy efficiently predicted an immunodominant viral T-cell epitope. PMID- 14550814 TI - Relapse or progression after hematopoietic cell transplantation using nonmyeloablative conditioning: effect of interventions on outcome. AB - OBJECTIVE: This study analyzes the effect of interventions aimed at reinducing remissions in patients with relapse or progression of malignant disease following allogeneic hematopoietic cell transplantation (HCT) using nonmyeloablative conditioning. METHODS: We performed a retrospective analysis of 81 instances of relapse or progression occurring among 224 patients given HCT as treatment of their hematologic malignancies. All patients received conditioning with 2 Gy total-body irradiation with or without fludarabine and with postgrafting immunosuppression with mycophenolate mofetil and cyclosporine. RESULTS: Overall survival of patients after relapse or progression was 36%. Fifteen of the 81 patients were given no interventions. Three of these 15 (20%) patients are alive with disease while 12 died with disease progression. Sixty-six patients (81%) received interventions, including withdrawal of immunosuppression (n=32), donor lymphocyte infusions (n=13), or chemotherapy (n=21). Twenty of the 66 (30%) are alive, 5 in complete remission, 4 in partial remission, 1 with stable and 10 with progressive disease. The overall response rate to intervention was 27%. Forty-six (70%) of the patients given interventions died, mainly due to relapse/progression. Patients not receiving interventions had a 1-year survival estimate of 15% compared to 41% in patients given interventions. Factors associated with survival in patients given intervention were disease response (p=0.002), disease category (p=0.001), and time to relapse from transplantation (p=0.0005). CONCLUSIONS: While the overall prognosis of patients relapsing or progressing after nonmyeloablative HCT is poor, interventions such as the combined use of immunotherapy and chemotherapy can improve patient survival. PMID- 14550815 TI - Allogeneic hematopoietic cell transplantation for patients with high-risk acute lymphoblastic leukemia in first or second complete remission using fractionated total-body irradiation and high-dose etoposide: a 15-year experience. AB - OBJECTIVE: The rationale for this retrospective study was to identify the long term overall and event-free survival, relapse, and treatment-related mortality rates of high-risk pediatric and adult first (CR1) and second remission (CR2) patients with acute lymphoblastic leukemia (ALL) who were treated with a single preparatory regimen consisting of fractionated total-body irradiation (FTBI) and high-dose etoposide (VP-16) prior to allogeneic hematopoietic cell transplantation. PATIENTS AND METHODS: Over a 15-year period at Stanford University Medical Center, 85 consecutive high-risk pediatric (up to age 17 years; n=41) and adult (age 18-55 years; n=44); patients with leukemia (ALL) in CR1 (n=55) and CR2 (n=30) received HLA-matched sibling allogeneic bone marrow or peripheral blood progenitor grafts after being treated with FTBI (1320 cGy) and high-dose VP-16 (60 mg/kg) as their preparatory regimen. The majority of patients transplanted in CR1 (n=45) had high-risk features, including age above 30 years, white blood cell count at presentation exceeding 25000/microL, extramedullary disease, need for more than 4 weeks of induction chemotherapy to achieve CR, or high-risk chromosomal translocations. Most patients transplanted in CR1 were adults (n=39), whereas patients in CR2 were primarily children or adolescents (n=25). RESULTS: The 10-year Kaplan-Meier estimates of relapse were significantly (p=0.05) lower in CR1 patients (15%+/-10%) than in CR2 patients (33%+/-20%). Relapse was the most common cause of treatment failure in patients transplanted in CR2. There was a significantly (p=0.05) higher rate of chronic graft-vs-host disease in CR1 (32%+/-14%) compared with CR2 (9%+/-11%) patients; however, overall survival for patients transplanted in CR1 (66%+/-14%) was comparable (p=0.67) to that of patients transplanted in CR2 (62%+/-19%). Event-free survival rates also were similar (p=0.53) between CR1 (64%+/-14%) and CR2 (61%+/-18%) patients. Treatment-related mortality rates were equivalent (p=0.51) between CR1 and CR2, as well as between Philadelphia chromosome (Ph) positive (Ph(+))and Ph( ) (p=0.23) ALL patients. CONCLUSION: Overall, FTBI/VP-16 is a highly effective preparatory regimen that provides durable remissions for patients receiving allogeneic hematopoietic cell transplantation for high-risk ALL in CR1 or CR2. PMID- 14550816 TI - Hyperhomocysteinemia in heart transplantation: from bench to bedside. PMID- 14550817 TI - Resistance exercise training and alendronate reverse glucocorticoid-induced osteoporosis in heart transplant recipients. AB - BACKGROUND: Immunosuppression therapy with bolus glucocorticoids causes regional osteoporosis in the axial skeleton of heart transplant recipients (HTR). No preventive strategy is generally accepted for steroid-induced bone loss. METHODS: To determine the efficacy of an anti-osteoporosis regimen that combined a bisphosphonate agent (alendronate sodium) with the osteogenic stimulus of mechanical loading, 25 HTRs were randomly assigned either to a group that received alendronate (10 mg/day) for 6 months (ALEN; n = 8), a group that received alendronate (10 mg/day) and performed specific resistance exercises for 6 months (ALEN + TRN; n = 8) or to a non-intervention control group (CONTR; n = 9). Alendronate was initiated at 2 months after transplantation. Bone mineral density (BMD) of the total body, femur neck and lumbar spine (L-2 and L-3) was measured by dual-energy X-ray absorptiometry before and 2, 5 and 8 months after transplantation. Resistance training consisted of lumbar extension exercise (MedX) performed 1 day/week and 8 variable resistance exercises (MedX) performed 2 days/week. RESULTS: Pre-transplantation BMD values did not differ among the 3 groups. BMD of the total body, femur neck and lumbar vertebra were significantly decreased below baseline at 2 months after transplantation in CONTR (-2.6 +/- 0.9%, -5.1 +/- 1.8%, -12.5 +/- 4.2%, respectively), ALEN (-2.8 +/- 0.8%, -5.3 +/- 1.6%, -12.0 +/- 3.9%) and ALEN + TRN groups (-2.7 +/- 1.0%, -5.6 +/- 2.1%, -11.2 +/- 3.7%). CONTR had further significant losses of BMD after 3 and 6 months. ALEN had no further regional BMD losses after initiation of alendronate therapy. ALEN + TRN restored BMD of the whole body, femur neck and lumbar vertebra to within 0.9%, 2.1%, and 3.4% of pre-transplantation levels, respectively. CONCLUSIONS: Resistance exercise plus alendronate was more efficacious than alendronate alone in restoring BMD in HTRs. Our results indicate that anti-osteoporosis therapy in this population should include both an anti-resorptive agent as well as an osteogenic stimulus, such as mechanical loading. PMID- 14550818 TI - Heart transplantation in patients with diabetes mellitus in the current era. AB - BACKGROUND: In this study, we used a single-institution database to examine the risks of heart transplantation in patients with diabetes mellitus (DM). METHODS: Recipients 18 years and older who underwent cardiac transplantation from July 1994 to December 2000 were reviewed; 101 consecutive patients with insulin dependent diabetes mellitus and non-insulin-dependent diabetes mellitus were compared with 244 patients who did not have DM and who received standard donor hearts. Survival, renal function (serum creatinine concentration), development of transplant coronary artery disease (TCAD), severe rejection, and infection (requiring hospitalization) were analyzed. RESULTS: Patients with DM were older (mean age, 57.1 years vs 51.4 years), had greater body mass index (mean, 26.7 vs 24.1 kg/m(2), p < 0.02), and more commonly had ischemic cardiomyopathy (58% vs 43%, p = 0.02). We found a trend toward decreased survival for those with DM at 1 year (85.1% vs 90.9%; p = 0.12). Five-year survival was 81.6% for both groups. Mean follow-up time was 4.1 years. Infection rate within 3 months was greater among those with DM (14% vs 3%, odds ration = 5.09; 95% confidence interval, 1.59 16.23). Freedom from infection at 4 years was 71.0% for patients with DM and 85.0% for those without DM (p = 0.02). Freedom from rejection at 4 years was similar (70.6% vs 73.6%, p = 0.69). At 4 years, transplant coronary artery disease (TCAD)-free survival was 69.5% for those with DM and 81.6% for those without (p = 0.23). Mean serum creatinine concentration at 4 years after transplant was 1.5 mg/dl in patients with DM (vs 1.4, p = 0.28). Multivariate analysis showed increased baseline creatinine level as a significant risk factor for survival and showed pre-transplant ischemic cardiomyopathy as a risk factor for TCAD in both groups. Body mass index >30 was a significant risk factor for survival among patients with DM. CONCLUSION: We found an increased risk of serious infections in patients with DM, particularly in the early post-operative period. Careful consideration of obesity and renal function during evaluation of candidacy is indicated. PMID- 14550819 TI - Longer-term risks associated with 10-year survival after heart transplantation in the cyclosporine era. AB - BACKGROUND: Long-term survival after heart transplantation is common in the cyclosporine era. However, there are few data documenting pre-transplant/peri operative factors predictive of truly long-term survival (>10 years). The purpose of this study is to identify factors associated with 10-year survival after heart transplantation. METHODS: Our study population included 197 adults who survived >6 months and died <10 years after heart transplant (medium-term group) and 140 adults who survived >10 years after heart transplant (long-term group) between December 1980 and May 2001. A comparison was done between the two groups and we used multivariate analysis to identify which factors predicted 10-year survival. RESULTS: The long-term group had younger recipient and donor age, lower recipient body mass index at transplant, shorter waiting time and lower percentages of ischemic etiology/male recipient/non-white recipient. Kaplan-Meier plots of freedom from graft coronary artery disease and malignancy showed later onset patterns in the long-term group compared with the medium-term group. Multivariate analysis showed that white recipient, younger recipient and lower recipient body mass index at heart transplant were factors significantly associated with 10-year survival. CONCLUSIONS: Several pre-transplant/peri-operative factors were associated with survival beyond 10 years after heart transplantation. Stratified/tailored strategies based on these factors may be helpful to attain longer-term survival of recipients with higher risks. PMID- 14550820 TI - Mycophenolate mofetil significantly reduces leukocyte graft infiltration after heterotopic cardiac transplantation in a rat model: comparative study with cyclosporine and FK 506. AB - BACKGROUND: The purpose of the study was to evaluate the effects of cyclosporine (CsA), FK 506 and mycophenolate mofetil (MMF) on graft-infiltrating leukocytes (CD4, CD8, CD11a, CD18) after cardiac transplantation in rats. METHODS: Three hundred forty animals were transplanted and randomly divided into 4 groups: CsA, 3 mg/kg/d (n = 74); MMF, 40 mg/kg/d (n = 96); FK 506, 0.3 mg/kg/d (n = 96); and a control group receiving no immunosuppressive therapy (n = 74). Three or 4 animals from each group were killed at intervals of 1 to 4 days up to Day 60. Immunohistochemistry was performed using monoclonal antibodies (MAb) against CD4, CD8, CD11a and CD18. Positively stained cells were analyzed in the perivascular space (PVS) of intra- and epicardial arteries. Statistical analysis was performed using area-under-the-curve assessment with an extended t-test. RESULTS: CsA and FK 506 reduced the presence graft-infiltrating leukocytes (CD4, CD8, CD11a, CD18) in the PVS of intra- and epicardial arteries when compared with control animals. MMF therapy resulted in a further significant reduction in infiltrating leukocytes when compared with the 2 calcineurin inhibitors. MMF had a faster onset of action than the calcineurin inhibitors. CsA and FK 506 required 12 to 20 additional days to achieve the reducing effect of graft infiltration seen in MMF treated animals. CONCLUSION: MMF possesses potent infiltration-blocking properties and its application leads to a greater reduction of cellular infiltration in the course of transplant rejection when compared with calcineurin inhibitors. PMID- 14550821 TI - Everolimus in de novo cardiac transplantation: pharmacokinetics, therapeutic range, and influence on cyclosporine exposure. AB - We evaluated exposure, safety, and efficacy data from an international Phase 3 trial of everolimus in de novo heart transplantation to characterize the longitudinal pharmacokinetics of everolimus and cyclosporine and to identify a therapeutic concentration range for everolimus. We randomized 634 patients to receive either 0.75 mg everolimus twice daily, 1.5 mg everolimus twice daily, or azathioprine in addition to corticosteroids and cyclosporine. At 8 visits during the first 6 months after transplantation, we obtained 2,328 everolimus trough levels (Cmin) and 129 area-under-the-curve (AUC) profiles over the dosing interval in patients treated with everolimus; we collected 3,258 cyclosporine trough concentrations and 174 profiles in all 3 treatment arms. We used median effect analysis to characterize exposure-response associations between everolimus average Cmin vs freedom from biopsy-confirmed acute rejection; maximum cholesterol, low density lipoprotein, triglyceride, and creatinine levels; and minimum leukocyte and platelet counts. Everolimus Cmins averaged 5.2 +/- 3.8 ng/ml and 9.4 +/- 6.3 ng/ml at the lower and upper dose levels. A 17% underproportionality was noted in Cmins; however, peak exposure and AUC were consistent with dose proportionality. Everolimus exposure was stable during the 6 month period. Interindividual variability was 37% for AUC and 40% for Cmin. The latter parameter was not influenced to a clinically relevant extent by sex, age, or weight. The Cmin was well correlated with AUC (r2 = 0.81). Everolimus Cmin was significantly related to freedom from rejection (p = 0.02) with 3 ng/ml being an informative lower threshold for efficacy. Thrombocytopenia, defined as <75 x 10(9)/liter, was related significantly to Cmin (p = 0.03); however, the incidence in this study was too low to establish an upper end for the therapeutic range. Lower doses of cyclosporine (by 15% to 19%) were used in patients treated with everolimus to achieve cyclosporine Cmins and AUCs similar to those in patients treated with azathioprine. Everolimus exposure was dose proportional and stable during the first 6 months after transplantation. Interindividual pharmacokinetic variability was high but not influenced by common demographic covariates. We observed a significantly increased risk of acute rejection at everolimus trough levels <3 ng/ml, which constitutes the lower therapeutic concentration limit when everolimus is used with conventionally dosed cyclosporine. Everolimus-related adverse events were manageable up to the highest troughs (22 ng/ml) observed in this population. We could not derive a precise upper therapeutic concentration limit from these data. PMID- 14550822 TI - Atrial tachyarrhythmias and permanent pacing after pediatric heart transplantation. AB - BACKGROUND: Atrial tachyarrhythmias have been reported in as high as 50% of adult heart recipients. Limited information is available on arrhythmias in pediatric transplant patients. Our objective was to determine the prevalence and significance of atrial tachyarrhythmias and permanent pacing following pediatric heart transplantation. METHODS: A retrospective review of the medical records, electrocardiograms, and Holter recordings of all consecutive patients following heart transplantation at Children's Hospital, Boston (n = 104) and Lucile Packard Children's Hospital, Stanford (n = 123) was performed. The study group consisted of 227 patients with a median age at transplant of 10.2 yrs (1 day-23.3 yrs). RESULTS: Atrial tachyarrhythmias occurred in 32 patients (14%) at a median of 15 days post-transplant (1 day-9.2 yrs) and included atrial flutter (n = 13), atrial fibrillation (n = 7), ectopic atrial tachycardia (n = 5), atrioventricular reciprocating tachycardia or atrioventricular node reentry (n = 5), and other (n = 2). Atrial flutter was the only tachyarrhythmia associated with allograft rejection (6/13 atrial flutter vs. 0/7 atrial fibrillation vs. 0/5 ectopic atrial tachycardia, p = 0.03). Patients with atrial fibrillation had a 2.5 fold (95%CI 1.7-3.5) higher risk of death or retransplant compared to patients without atrial fibrillation. Ectopic atrial tachycardia tended to occur in younger recipients compared to atrial fibrillation and flutter (2.7 yrs vs 18.6 yrs and 8.5 yrs respectively, p = 0.06) and was associated with a benign clinical course. There was no association between atrial tachyarrhythmias and graft ischemic time, surgical technique, or coronary artery disease. Pacemakers were required in 12 patients (5.2%), 7 with sinus node dysfunction and 5 for intermittent complete atrioventricular block. There was no consistent association between the need for permanent pacing and coronary artery disease, rejection, or surgical technique. CONCLUSIONS: Atrial tachyarrhythmias and permanent pacing were uncommon in this cohort of pediatric heart transplant recipients. Association with cardiac rejection, clinical course, and mortality varied depending on the tachyarrhythmia mechanism. PMID- 14550823 TI - Benefits of cardiac resynchronization therapy in outpatients with indicators for heart transplantation. AB - BACKGROUND: Despite several new advances in the treatment of end-stage congestive heart failure (CHF) the number of patients moving onto the heart transplant list continues to rise. Recently, cardiac resynchronization therapy (CRT) has become a Food and Drug Administration (FDA)-approved therapy for advanced CHF. Currently, there are no available data on CRT in cardiac transplant candidates. METHODS: To determine the effects of CRT on potential transplant candidates we retrospectively reviewed patients (n = 34) enrolled in resynchronization trials at our center who met accepted criteria for transplantation. Ventricular function, oxygen uptake and New York Heart Association (NYHA) class were compared at baseline and after 6 months of active therapy for each patient. RESULTS: CRT reduced QRS duration from 178 +/- 29 to 143 +/- 17 msec (p < 0.0001) and NYHA class from 3.1 +/- 0.3 to 1.8 +/- 0.7 (p < 0.0001). Ejection fraction (EF) increased from 18.4 +/- 5.1 to 25.1 +/- 8.4% (p < 0.0001) and maximum VO(2) increased from 11.9 +/- 1.8 to 15.3 +/- 3 ml/kg/min (p < 0.0001). Only 2 of the initial 34 patients still met the criteria for transplantation at 6 months. CONCLUSIONS: CRT improves ventricular function, oxygen uptake and NYHA class in ambulatory patients who have a QRS of >130 msec and might be considered candidates for cardiac transplantation. Transplantation can be prevented or deferred in the majority of patients. Outpatients with chronic CHF and a widened QRS should be evaluated for CRT before being listed for cardiac transplantation. PMID- 14550824 TI - Timing of cardiac transplantation in patients with heart failure receiving beta adrenergic blockers. AB - BACKGROUND: Previous work shows that patients with heart failure patients who have peak oxygen consumption (VO2 peak) >14 ml/kg/min do not derive a survival benefit from cardiac transplantation. However, this was shown before beta-blocker therapy for patients with systolic heart failure became common, and beta-blockers improve survival in patients with heart failure without changing VO(2) peak. Our purpose was to re-evaluate the utility of VO(2) peak >14 ml/kg/min as an indicator of the need for cardiac transplantation in patients with heart failure who are taking beta-blockers. METHODS: Actuarial, hemodynamic, and exercise ventilatory data were collected from 540 patients with heart failure, 256 of whom were taking beta-blockers. We tracked death and cardiac transplantation. We stratified the percentage of patients event-free 1 and 3 years after VO(2) peak study by their VO(2) peak and beta-blocker status, and compared 1- and 3-year post-transplant survival (United Network of Organ Sharing [UNOS] data). We also compared total mortality for the patients with heart failure as stratified by beta-blocker stats and VO(2) peak (excluding the 42 who underwent transplantation) with UNOS post-transplant survival. RESULTS: Patients with heart failure who were receiving beta-blockers and whose VO(2) peak was > or =12 ml/kg/min had greater 1- and 3-year event-free survival rates (95% confidence intervals, 92.6%-96.6% and 85.8%-96.0%) than did post-transplant patients (83.9% 86.3% and 75.4%-76.6%). However, in patients with heart failure not taking beta blockers, VO(2) peak <14 ml/kg/min was associated with worse 3-year survival (38.9 - 62.1%) than that for post-transplant patients. Excluding the 42 patients with heart failure in our study who underwent transplantation and then evaluating survival of the remaining patients with heart failure (not event-free survival) did not substantially change these results. CONCLUSIONS: Patients with heart failure who are receiving beta-blockers do not derive a survival advantage at 1 and 3 years after cardiac transplantation if VO(2) peak is > or =12 ml/kg/min. Patients not taking beta-blockers whose VO(2) peak is <14 ml/kg/min have superior survival with cardiac transplantation. PMID- 14550825 TI - Modeling the effects of functional performance and post-transplant comorbidities on health-related quality of life after heart transplantation. AB - BACKGROUND: Health-related quality of life and functional performance are important outcome measures following heart transplantation. This study investigates the impact of pre-transplant functional performance and post transplant rejection episodes, obesity and osteopenia on post-transplant health related quality of life and functional performance. METHODS: Functional performance and health-related quality of life were measured in 70 adult heart transplant recipients. A composite health-related quality of life outcome measure was computed via principal component analysis. Iterative, multiple regression based path analysis was used to develop an integrated model of variables that affect post-transplant functional performance and health-related quality of life. RESULTS: Functional performance, as measured by the Karnofsky scale, improved markedly during the first 6 months post-transplant and was then sustained for up to 3 years. Rejection Grade > or =2 was negatively associated with health-related quality of life, measured by Short Form-36 and reversed Psychosocial Adjustment to Illness Scale scores. Patients with osteopenia had lower Short Form-36 physical scores and obese patients had lower functional performance. Path analysis demonstrated a negative direct effect of obesity (beta = - 0.28, p < 0.05) on post-transplant functional performance. Post-transplant functional performance had a positive direct effect on the health-related quality of life composite score (beta = 0.48, p < 0.001), and prior rejection episodes grade > or =2 had a negative direct effect on this measure (beta = -0.29, p < 0.05). Either directly or through effects mediated by functional performance, moderate-to severe rejection, obesity and osteopenia negatively impact health-related quality of life. These findings indicate that efforts should be made to devise immunosuppressive regimens that reduce the incidence of acute rejection, weight gain and osteopenia after heart transplantation. PMID- 14550826 TI - Determinants of quality of life changes among long-term cardiac transplant survivors: results from longitudinal data. AB - BACKGROUND: Cross-sectional analyses have identified significant associations between quality of life (QOL), and comorbidities and adverse effects in cardiac transplant recipients. However, little is known about factors that influence changes in QOL over time. This study examines both cross-sectional and longitudinal data from long-term survivors to identify factors that affect differences in QOL among recipients and individual changes in QOL during a 1-year period. METHODS: Self-selected enrollees completed questionnaires, including QOL scales, at 3-month intervals. Repeated measures multiple regression analysis was used to examine the association between the QOL scales and comorbidities, adverse effects, and compliance measures, controlling for other factors. RESULTS: We included 569 participants in the analysis, with a mean time since transplantation of 8.6 years. Cross-sectional results showed that the number of comorbidities, treatment non-compliance, and several adverse effects were associated with low QOL. In longitudinal results, waiting to take medications and taking less medication because of lifestyle restrictions were associated with decreases in QOL over time. Hair loss, changes in face shape, and decreased sexual interest or ability also had the largest adverse effects on changes in QOL. CONCLUSIONS: These findings provide new opportunities for interventions to address factors related to decreases in QOL. Clinicians should actively solicit information about compliance with medication regimens. In addition, information about the adverse effects of medications should be considered when making therapeutic decisions. PMID- 14550827 TI - Multidrug-resistant tuberculosis in a lung transplant recipient. AB - Tuberculosis infection has been a relatively rare complication after lung transplantation. However, as more countries in which Mycobacterium tuberculosis infection remains endemic embark on lung transplant programs, the occurrence of multidrug-resistant tuberculosis after transplantation is a genuine threat. We report the first case of multidrug-resistant tuberculosis in a double-lung transplant recipient who probably acquired the disease from the donor. We discuss the problems in clinical management of post-transplant tuberculosis infection and of drug-resistance. PMID- 14550828 TI - Primary adenocarcinoma in a donor lung: evaluation and surgical management. AB - We report a case of primary non-small cell cancer diagnosed in the donor lung 33 months after bilateral pulmonary transplantation. The tumor was treated surgically. A lingular sparing left upper lobe bisegmental lung resection was performed. PMID- 14550829 TI - Off-pump coronary bypass surgery for left main coronary artery stenosis 10 years after heart transplantation: case report. AB - A patient with asymptomatic left main coronary artery stenosis 10 years after heart transplantation was treated successfully with off-pump coronary bypass surgery using both mammary arteries. New advances in bypass surgery may decrease the risk of revascularization in cardiac transplant recipients. PMID- 14550831 TI - Serotonin and norepinephrine involvement in efferent pathways to the urethral rhabdosphincter: implications for treating stress urinary incontinence. AB - Stress urinary incontinence (SUI), the most common form of incontinence, continues to be a largely underdiagnosed problem that imposes large financial and quality-of-life burdens on many women but has few treatment options. Ongoing animal and early human studies have shown that monoamine neurotransmitters play key roles in controlling urethral storage and micturition reflexes. Motor neurons found in the Onuf nucleus of the sacral spinal cord control urethral function, and have several unique properties that distinguish them from other motor neurons. First, the neurons are uniformly smaller than other surrounding motor neurons and have bundled dendrites, allowing strong synchronous activation or inhibition. Second, the neurons demonstrate unique neurochemical profiles. Unlike neurons in surrounding areas, the motor neurons of the Onuf nucleus have dense populations of noradrenergic and serotonergic terminals. Animal studies have shown that alpha1-adrenoceptors and serotonin (5-hydroxytryptamine [5-HT]) receptors in the Onuf nucleus facilitate sphincter contraction. Agonists that stimulate these receptors facilitate the guarding or incontinence reflex, whereas antagonists that block the receptors inhibit this reflex. Therefore, boosting the effects of 5-HT and norepinephrine (NE) to enhance sphincter activity could be clinically promising for improving the symptoms of SUI. Importantly, the activity of the sphincter neurons can be increased pharmacologically during urine storage without interfering with bladder-sphincter synergy. Administering the 5-HT/NE uptake inhibitor duloxetine facilitates sphincter contraction during bladder filling but not during bladder contraction in micturition. This unique effect of duloxetine may be maintained by the selective neuromodulatory effects of 5-HT and NE on activation of sphincter motor neurons by the neurotransmitter glutamate. Prolonging the effect of naturally released NE and 5-HT with duloxetine could augment the body's normal processes for controlling urine storage and micturition. Early trials have demonstrated that duloxetine significantly reduces incontinence episodes and is well tolerated in the clinical setting. PMID- 14550832 TI - Serotonergic modulation of bladder afferent pathways. AB - Normal bladder function is based on activation and maintenance of a sophisticated reflex mechanism involving sympathetic, parasympathetic, and somatic control of the lower urinary tract. The spinal and supraspinal neuronal pathways involved can be modulated by activation or inhibition of neurons in the periphery, at the lumbosacral and thoracolumbar spinal levels, and at supraspinal regulatory sites. Activation of the primary afferent neurons that innervate the lower urinary tract is the first step on this reflex pathway. Under conditions in which bladder function is compromised, abnormal activity in these afferent neurons can induce changes in these circuits, resulting in bladder dysfunction. Control and modulation of afferent pathways is a recent focus for the development of novel treatments for lower urinary tract disorders. This review focuses on the central regulation of bladder function by central serotonergic modulation of sensory pathways. Modulation of this monoaminergic system has dramatic effects on bladder activity and can be a target for pharmacologic treatment of bladder disorders. PMID- 14550833 TI - Epidemiology and natural history of urinary incontinence in women. AB - Understanding the epidemiology (distribution and determinants) of urinary incontinence (UI), as well as its natural history is a very important issue. In this article, we discuss prevalence, incidence, natural history, and the variations that may be related to race and ethnicity. We focus on epidemiologic population comprising community-dwelling women who are not institutionalized. Our review clearly shows that there is a lack of advanced epidemiologic analyses. Variables that better characterize UI include frequency measure, quantity of urine loss, duration, type, and severity. These factors should be incorporated into basic study design so that more advanced and informative analyses may be conducted. PMID- 14550834 TI - Home electrical stimulation for urinary incontinence: a study of the diffusion ofa new technology. AB - The objective of this study was to investigate the extent to which physicians requested home electrical stimulation (ES) treatment for patients with urinary incontinence (UI). Home ES was requested by 429 general practitioners (GPs) and 147 gynecologists within 2 years (1992 to 1994) after the Norwegian National Insurance Service offered reimbursement for the procedure. For first-time requesters, cumulative plots of time to request were compared for GPs versus gynecologists. Characteristics of requesting GPs were compared with those of a randomly selected control sample of nonrequesting GPs, collected by postal questionnaire. A greater proportion of gynecologists than GPs prescribed home ES during the 2-year study period (42% vs 14%, P <0.001). Median time to request for ES was 5.8 months for gynecologists versus 8.6 months for GPs (P <0.01). A greater proportion of female GPs than male GPs requested the treatment (17% vs 13%, P <0.01). Sufficient information about and/or knowledge of treatment indications were claimed by 71% of requesting GPs versus 21% of controls (P <0.001). Altogether, 91% of requesters versus 62% of controls judged that home ES was effective or a good treatment alternative (P <0.001). Requesters (versus controls) prescribed more bladder training and estrogen for urge UI, more pelvic floor exercises and estrogen to treat stress UI, and more ES to treat both types of UI (all P <0.05). Gynecologists prescribed ES more often and earlier than GPs. Prescribers of ES were more proactive in treating UI in their practices than those who were not prescribers. PMID- 14550835 TI - Pharmacologic treatment of stress urinary incontinence: expectations for outcome. AB - Several classes of drugs have been investigated for their efficacy in treating stress urinary incontinence (SUI). Despite targeting the mechanisms known to be involved in maintaining continence, few have provided the desired benefits without causing significant adverse effects. Duloxetine, the newest drug to be extensively evaluated for SUI treatment, appears to be both safe and effective. None of the drugs tested thus far has proved to be curative, however. Treatment strategies therefore are aimed at improving the quality of life of patients and should take into account their individual needs and preferences. PMID- 14550836 TI - Childbirth-induced trauma to the urethral continence mechanism: review and recommendations. AB - To summarize the literature on immediate pelvic floor damage from childbirth and episiotomy, a MEDLINE search of English language articles published from 1983 to 2001 was performed. Vaginal delivery causes varying degrees of muscular, neuromuscular, and connective tissue damage. This damage may result in urinary and/or fecal incontinence. Routine midline episiotomy increases the risk of third and fourth-degree perineal lacerations, which may lead to fecal incontinence. Routine use of mediolateral episiotomy does not prevent urinary incontinence (UI) or severe perineal tears. It is possible to reduce the rate of mediolateral episiotomy to as low as 20% in primiparas without increasing the risk of anal sphincter damage. Control of obesity before delivery, as well as pelvic floor exercises and regular physical exercise both before and after delivery, seem to reduce the risk of postpartum UI. PMID- 14550837 TI - The role of estrogens in female lower urinary tract dysfunction. AB - The urogenital tract and lower urinary tract are sensitive to the effects of estrogen and progesterone throughout adult life. Epidemiologic studies have implicated estrogen deficiency in the etiology of lower urinary tract symptoms that occur after menopause. Although the role of estrogen replacement therapy in the management of postmenopausal urinary incontinence (UI) remains controversial, its use in the treatment of women with urogenital atrophy is now well established. This review summarizes recent evidence of the urogenital effects of hormone therapy, particularly emphasizing management of postmenopausal UI and recurrent lower urinary tract infections. Estrogen therapy alone has little effect in the management of urodynamic stress UI, although in combination with an alpha-adrenergic agonist, it may improve urinary leakage. Estrogen therapy may be of benefit for the irritative symptoms of urinary urgency, frequency, and urge UI, although this effect may result from reversal of urogenital atrophy rather than a direct action on the lower urinary tract. The role of estrogen replacement therapy in the treatment of women with recurrent lower urinary tract infections remains to be determined, although there is now some evidence that vaginal administration may be efficacious. Low-dose, vaginally administered estrogens have a role in the treatment of urogenital atrophy in postmenopausal women and appear to be as effective as systemic preparations. PMID- 14550838 TI - Trends toward less invasive treatment of female stress urinary incontinence. AB - Recent improvements in our understanding of the underlying pathophysiologic mechanisms responsible for stress urinary incontinence (SUI) in women have led to the development of innovative new surgical methods. Many are less invasive than previous techniques and appear to offer improved safety and shorter hospital stays, while maintaining the efficacy of traditional open incontinence surgery. Procedures using injectable periurethral bulking agents, laparoscopic colposuspension, and insertion of tension-free vaginal tape characterize this current trend toward less invasive surgical treatments. The increasing range of available procedures allows surgical treatment of SUI to be individualized for the patient. Women of diverse ages and levels of medical fitness can increasingly be offered a choice of safe, effective treatment for SUI. PMID- 14550847 TI - Genetic and pharmacological aspects of histamine H3 receptor heterogeneity. AB - Histaminergic H3 receptors modulate the release of neurotransmitters within the CNS and periphery. Ligands for these receptors have potential clinical utility in a variety of disease states. However, the pharmacological characteristics of these receptors have been enigmatic for more than a decade because of the diversity of pharmacological effects observed with the limited number of heretofore-available compounds. Recent cloning of the H3 receptor has revealed interspecies differences in the protein sequences in key regions, the existence of splice variants that differ in composition between species, and potential differences in signal transduction processes between either different tissues and/or species. This review attempts to summarize these findings within the context of the molecular biological and pharmacological data accumulated to date. Also, we suggest a nomenclature strategy to reduce potential confusion that has arisen from different naming systems used by various investigators. While some facets of this genetic and pharmacological diversity help to rationalize various aspects of H3 receptor heterogeneity, there remains an insufficient repertoire of selective ligands, assays, or other measures to completely resolve all components of this diversity. The promise of newly available tools to further explore H3 receptor function may provide the insight to bring the promised clinical potential of H3 receptor ligands to realization. PMID- 14550848 TI - Evaluation of the nocturnal levels of urinary biogenic amines in men exposed overnight to 50-Hz magnetic field. AB - The aim of this study was to determine whether exposure to magnetic fields might affect human health and to look for possible effects of acute exposure (9 hours) to 50-Hz magnetic fields (10 microT) on the urinary concentration of biogenic amines. Thirty-two young men (20-30 years old) were divided into two groups (sham exposed and exposed group) of 12 to 16 subjects each. All subjects participated in two 24-hour experiments to evaluate the effects of both continuous and intermittent exposure to magnetic fields. The subjects were exposed to the magnetic field from 2300 to 0800, while lying down. Total urine (from 2300 to 0800) was collected at 0800. The results (expressed as a ratio of biogenic amine excretion to creatinine excretion (nmol/mmol)) did not differ significantly between sham-exposed and exposed men for any of the parameters measured: adrenaline, noradrenaline, dopamine, dihydroxyphenylalanine, 3,4 dihydroxyphenylacetic acid, homovanillic acid and 5-hydroxyindoleacetic acid. These results suggest that nocturnal exposure to either continuous or intermittent 50-Hz magnetic fields of 10 microT does not affect, at least under our experimental conditions, the nocturnal excretion of biogenic amines in healthy young men. PMID- 14550849 TI - Endothelial modulation and tolerance development in the vasorelaxant responses to nitrate of rabbit aorta. AB - We investigated the endothelial modulations in nitrate tolerance in isolated rabbit aorta. Nitrate tolerance was induced by a 72-h treatment with transdermal nitroglycerin (NTG, 0.4 mg/h) in conscious rabbits, which was verified by a 20 fold increase in the EC50 values [NTG tolerance (6.1 +/- 0.8) x 10(-7) M vs control (3.0 +/- 0.6) x 10(-8) M]. The relaxations to NTG in tolerant and nontolerant aortic strips were enhanced when their endothelia were denuded [E( )]. In the presence of endothelium [E(+)], NTG-tolerant vessels were not tolerant to acetylcholine (ACh), which can release endothelial nitric oxide (NO), exogenous NO or 8-bromo (Br)-cGMP. In NTG-tolerant and nontolerant vessels with endothelium, concentration-response curves for NO were the same as those in endothelium-absent tolerant vessels. In both NTG-tolerant and nontolerant vessels, treatment with superoxide dismutase (SOD, 20 units/ml), an O2-. scavenger, unaffected the responses to NTG reduced in the presence of endothelium, but treatment with NG-nitro-L-arginine methyl ester (L-NAME, 10(-4) M), an NO synthase (NOS) inhibitor, reversed these reductions. Thus, our data did not indicate that an increased endothelial superoxide O2-. production contributes to nitrate tolerance. Our study suggested that (i) an impaired biotransformation process from NTG to NO is responsible for the occurrence of nitrate tolerance and (ii) vascular response to NTG enhanced by endothelial removal is related to blocked endothelial NO release. PMID- 14550850 TI - Behavioral effects of acute and chronic triazolam treatments in albino rats. AB - Previous behavioral studies on triazolam (TZ), which are small in number, could only speculate about tolerance to the anxiolytic effect of TZ, as the experiments did not cover sufficient time (of 4 to 7 days) for tolerance to develop. Therefore longer time for chronic TZ administration is used. We investigated the effects of TZ on motor activity and exploratory behavior using plus maze and open field. Three experiments were conducted. In the first, five groups of rats were acutely treated with different doses of TZ (0.25 mg/kg-4.0 mg/kg). In the second set of experiments, rats were treated chronically with a single daily dose of TZ (started with 0.25 mg/kg and increased by time to 1.0 mg/kg) for 5 weeks (representing clinical use). In the third, rats were treated chronically with three daily doses of TZ (started with 0.25 mg/kg and increased by time to 0.5 mg/kg) for 20 days (mimicking drug abuse). Acute TZ administration produced dose dependent anxiolytic effects and a decrease in motor activity with higher doses. Chronically treated rats, either once daily or three times daily doses, showed tolerance to both anxiolytic and sedative effects of TZ. It may be concluded that tolerance to the anxiolytic and sedative effects of TZ would develop after chronic administration either with clinical use or its abuse. PMID- 14550851 TI - Transcriptional regulation of fosl-1 by licorice in rat Clone 9 cells. AB - Licorice is a commonly used herbal medicine for treatment of liver disorders. Its biological activities have been widely studied. However, little information on its transcriptional regulation has been reported. In the present study, the effect of an aqueous extract of licorice on the gene expression in rat liver cells (Clone 9) was investigated. The results show the expression of GST-pi, DT diaphorase, PAI-1, fosl-1 and uPAR were over two-fold increased. Northern blot analysis revealed that the over-expression of these genes was concentration dependent (0.25-3 mg/ml) but the temporal expression profile (8-48 h) of each individual gene varied. The over-expression of fosl-1 could be related to the event in the induction process leading to the expression of GST-pi, DT diaphorase, uPAR and PAI-1 through AP-1. Induction of the over-expression of GST pi and DT-diaphorase genes may contribute to the hepatoprotective properties of licorice whereas activation of uPAR and PAI-1 together with down-regulation of TIMP-3 suggest a role of LE in the regulation of cell mobility. PMID- 14550852 TI - Potential mechanisms responsible for chlorotriazine-induced alterations in catecholamines in pheochromocytoma (PC12) cells. AB - Chlorotriazines interact with undifferentiated PC12 cells in vitro to modulate catecholamine synthesis and release, but the mechanism(s) responsible for this effect had not been determined. In this study we evaluated the effect of atrazine, simazine and cyanazine on the protein expression of the enzymes responsible for the synthesis of dopamine [tyrosine hydroxylase (TH)] and norepinephrine [dopamine-beta-hydroxylase (DbetaH)]. We also examined the possible intracellular pathway associated with chlorotriazine-induced changes in catecholamine synthesis and release. Incubating PC12 cells in the presence of 100 microM atrazine and simazine decreased intracellular dopamine (DA), norepinephrine (NE) concentration and NE release, and the protein expression of TH (approximately 20%) and DbetaH (approximately 50 and 25%, respectively) after 12-24 h exposure. In contrast, cyanazine (100 microM) stimulated intracellular and released NE concentration, and the protein expression of TH (approximately 20%) and DbetaH (approximately 225%) after 12-36 h exposure. Simultaneous exposure to the essential TH co-factors (iron and tetrahydrobiopterine) was ineffective in altering cellular DA. Agents known to enhance TH and DbetaH transcription, phosphorylation or activity (e.g., 8-bromo cAMP, forskolin or dexamethasone) reversed the inhibitory effects of atrazine and simazine on the NE. Again, in contrast to atrazine and simazine, cyanazine attenuated catecholamine-depleting effect of alpha-Methyl-p-tyrosine (alphaMpT) on NE. Both DA and NE synthesis can be altered by the chlorotriazines and suggest these occur via an alteration of the synthetic enzymes TH and DbetaH. PMID- 14550853 TI - Naloxone potentiates treadmill running-induced increase in c-Fos expression in rat hippocampus. AB - The expression of c-Fos is induced by a variety of stimuli and is sometimes used as a marker for increased neuronal activity. In the present study, the effect of treadmill running on c-Fos expression in the hippocampus and the involvement of opioid receptors were investigated via c-Fos immunohistochemistry. It was shown that c-Fos expression in the CA1 region, the CA2 and CA3 regions, and the dentate gyrus of the hippocampus was significantly increased by treadmill running and naloxone, a nonselective opioid receptors antagonist, treatment enhanced treadmill exercise-induced increase of hippocampal c-Fos expression. Base on the present results, it can be suggested that treadmill running increases hippocampal neuronal activity and that endogenous opioids curtail the exercise-induced increase. PMID- 14550854 TI - Actions of insulin-like growth factor binding protein-5 (IGFBP-5) are potentially regulated by tissue kallikrein in rat brains. AB - Tissue kallikrein (EC 3.4.21.35) that hydrolyzes kininogen and releases a physiologically active peptide, kinin, is found in rat brains. Insulin-like growth factor binding proteins (IGFBPs) that modulate IGF actions are also expressed in a variety of tissues including rat brains, and one of the major IGFBPs expressed in brain is known to be IGFBP-5, which is reported to be hydrolyzed in vitro by prostate-specific antigen (PSA) and gamma-nerve growth factor (gamma-NGF), both of which belong to the member of the kallikrein gene family. This study was designed to determine whether or not kallikrein has a potential to hydrolyze IGFBP-5 and their topographic proximity was investigated in rat brain using double immunohistochemical staining method. Immunohistochemically, IGFBP-5 positive cells were numerous and widespread in the cerebral cortex and belonged to neurons in the cell configuration. IGFBP-5 positive cells were negative for S-100 protein and were positive for betaIII tubulin, confirming them to be neurons. In addition, kallikrein positive cells were virtually all IGFBP-5 positive cells. IGFBP-5 was clearly hydrolyzed by kallikrein with cleavage sites of Arg188-Met189 and Arg136-Arg137 of IGFBP-5. Therefore, there is a possibility that kallikrein plays an important role in brain physiology, specifically in the neurons by regulating the actions of IGFBP 5 and IGF. PMID- 14550855 TI - Metabolic and immunological responses associated with in vivo glucocorticoid depletion by adrenalectomy in mature Swiss albino rats. AB - The study is undertaken to determine the effect of adrenal corticosteroid depletion after adrenalectomy on carbohydrate, protein and fat metabolism as well as maturation and functional efficacy of the immunocompetent cells. Beside biochemical and hematological parameters, whether in vivo glucocorticoid depletion has any modulatory effects on splenic macrophage responses to bacterial challenge with regards to intracellular killing, nitric oxide release and cellular integrity, were determined. Major findings of our study indicate that blood glucose, urea and total inorganic phosphate levels showed a time dependent increase in adrenalectomized rats compared to control. Total glycogen content in liver was decreased gradually due to adrenal corticosteroid insufficiency. Hematological parameters like hemoglobin concentration, hematocrit value, total leukocyte count and differential count were also found to increase in the adrenalectomized group with respect to intact group. From the functional study of immunocompetent cells, intracellular killing capacity of splenic macrophages recovered from control and adrenalectomized rats after 10 and 20 days of adrenalectomy showed no significant alteration; however, the function of splenic macrophages recovered from rats after 30 days of adrenalectomy showed altered response. Nitric oxide released from splenic macrophages of adrenalectomized rats was less than that of control animal even after stimulation with lipopolysaccharide. DNA fragmentation assay showed a lesser degree of fragmentation of splenic macrophages obtained from adrenalectomized rats indicating, apoptotic death of cells in this group decreases. Adrenal corticosteroid insufficiency due to adrenalectomy interferes with metabolic and hematopoietic functions and modulates the development and maintenance of normal immunitary status, which in turn influences the inflammatory response. PMID- 14550856 TI - An overview of the recent trends in development of HPLC methods for determination of impurities in drugs. AB - An extensive survey of the literature published in various analytical and pharmaceutical chemistry related journals has been conducted and the high performance liquid chromatography (HPLC) methods which were developed and used for determination of process-related impurities in drugs have been reviewed. This review covers the time period from 1995 to 2001 during which around 450 analytical methods including all types of chromatographic and hyphenated techniques were reported. HPLC with UV detection was found to be the technique of choice for many workers and more than 200 methods were developed using LC-UV alone. A critical analysis of the reported data has been carried out and the present state-of-art of HPLC for determination of impurities of analgesic, antibiotic, anti-viral, anti-hypertensive, anti-depressant, gastro-intestinal and anti-neoplastic agents has been discussed. PMID- 14550857 TI - Determination of dissociation constants of labile drug compounds by capillary electrophoresis. AB - The utility of capillary electrophoresis (CE) for determination of the negative logarithm of dissociation constants (pK(a)) of labile compounds was investigated. In this study pyridinyl-methyl-sulfinyl-benzimidazoles (PMSB's), which have both an acidic and a basic pK(a), were selected as a first set of model drug compounds. This is a group of compounds that are known to degrade in aqueous solutions under neutral and acidic conditions which thus may impair their pK(a) determination when using common batch techniques based on spectrophotometry or potentiometry. An additional set of model drug compounds, benzenesulfonic acid phenethyloxy-phenyl esters (BSAP's), which are labile at high pH, were also studied. It is demonstrated that pK(a) values can be determined with high precision and accuracy by CE for both these sets of model compounds because decomposition products and impurities can be sufficiently separated from the main component. Based on the results in this study, a general strategy is proposed and discussed for determination of pK(a) for labile compounds. Key steps comprise use of a stabilizing sample diluent, injection by electromigration, short analysis time, and characterization of the main component by UV-Vis spectra. PMID- 14550858 TI - Determination of zolpidem hemitartrate by quantitative HPTLC and LC. AB - Two methods are described for the determination of zolpidem hemitartrate in presence of its degradation product. The first method was a TLC-UV densitometric one in which the mobile phase methanol: water (20:80) was used for developing the TLC plates. The R(f) of zolpidem hemitartrate was found to be 0.29+/-0.01 and that of its degradation product was 0.59+/-0.01. Linearity range was 0.5-4 microg/spot with mean recovery percentage (99.98+/-0.988)%. The second method was an HPLC method. HPLC was performed on a Bondapack C(18) column. The mobile phase was composed of a mixture of acetonitrile-0.01 M KH(2)PO(4) (40:60). The pH was adjusted to 3.5+/-0.1. Flow rate was 1.2 ml/min. Calibration graphs were linear in the range of 0.5-5 microg/ml with UV detection at 245 nm. Both methods have been successfully applied to pharmaceutical formulations. The results obtained were statistically compared with those obtained by applying the reported methods. PMID- 14550859 TI - The HPLC determination of propane-1,2,3-triyl trinitrite and impurities: (2RS)-3 hydroxypropane-1,2-diyl dinitrate and 2-hydroxypropane-1,3-diyl dinitrate in ointment. AB - A HPLC method for determination of propane-1,2,3-triyl trinitrate and impurities: (2RS)-3-hydroxypropane-1,2-diyl dinitrate and 2-hydroxypropane-1,3-diyl dinitrate ointment was developed. The conditions for good separation of constituents, while avoiding vehiculum interference were established. The results feature of high accuracy and good precision. For individual constituents R.S.D. is ranged from 0.7 to 9.9%, while recovery was 100.1% for propane-1,2,3-triyl trinitrate and 95.1-99.0% for impurities. It has been found that propane-1,2,3-triyl trinitrate used in medicine in the form of ointment contains such impurities which can be identified and quantified at relatively low concentrations of 70 ng ml(-1). PMID- 14550860 TI - Validation of the spectrophotometric determination of omeprazole and pantoprazole sodium via their metal chelates. AB - Spectrophotometric procedures for the determination of two irriversible proton pump inhibitors, omeprazole (OMZ) and pantoprazole (PNZ) sodium have been developed, the procedures are based on the formation of 2:1 chelates of both drugs with different metal ions. Pantoprazole sodium is quantified by a stability indicating procedure through chelation with iron (III) in aqueous-ethanol medium to form an orange chelate picked at 455 nm. The procedure retains its accuracy in presence of up to 70% of its degradate, sulfenic acid prepared by degrading the pure drug in borate buffer of pH 8 at 37 degrees C for 5 days. The colored chelates of OMZ in ethanol are determined spectrophotometrically at 411, 339 and 523 nm using iron (III), chromium (III) and cobalt (II), respectively. Regression analysis of Beer's plots showed good correlation in the concentration range of 15 95, 10-60 and 15-150 microml(-1) of pure OMZ using iron (III), chromium (III) and cobalt (II), respectively, and in the range of 30-300 microg ml(-1) of PNZ sodium using iron (III). The limits of detection are 0.22-3.65 microml(-1) while limits of quantitation range between 0.74 and 12.17 microg ml(-1). The optimum assay conditions are investigated and the recovery of the cited drugs from their dosage forms ranges from 97.2 to 100.3%. Good values of precision are obtained, intraday R.S.D. are 0.93-1.75% and the inter day R.S.D. are 0.51-3.29%. PMID- 14550861 TI - Determination of binary mixtures of analgesic and spasmolytic drugs in pure and dosage forms by derivative spectrophotometry. AB - Binary mixtures of dipyrone and pitophenone hydrochloride are assayed by zero crossing second- and third-derivative spectrophotometry and by ratio-spectra first- and second-derivative spectrophotometry. In the first method, calibration plots are linear at 266.5 and 302.5 nm (dipyrone, second derivative), and 257 and 286 nm (pitophenone second derivative) and 242 and 278.3 nm (dipyrone third derivative), and 228.5 and 300 nm (pitophenone, third-derivative). By the second method, lines of regression are linear at 235 and 262 nm (dipyrone, first derivative), and 229.5 and 288.5 nm (pitophenone, first-derivative), and 249.7 and 268 nm (dipyrone, second derivative), and 280.5 and 300 nm (pitophenone, second-derivative). In all methods calibration curves follow the Beer's law up to 40 microg/ml of each drug. LOD and LOQ values were calculated. The developed derivative spectrophotometric methods were applied to laboratory mixtures and to vials for these drugs. The procedures are simple, rapid, and did not require any preliminary separation or treatment of the samples. PMID- 14550862 TI - The control of nitrilotriacetic acid in edetic acid and its salts by liquid chromatography. AB - Two liquid chromatographic methods have been developed and validated for the determination of nitrilotriacetic acid (NTA) in edetic acid (EDTA) and its salts. In one method NTA and EDTA are separated on a polymer column without pretreatment, and are detected amperometrically using a glassy-carbon electrode. In the other method NTA and EDTA are complexed with ferric ions and the complexes separated on a porous graphite carbon stationary phase with ultraviolet detection at 215 nm. Both methods were sufficiently selected and sensitive to allow the control of NTA (0.1% m/m) in sample of EDTA and its salts. PMID- 14550863 TI - Simultaneous determination of paracetamol and chlorpheniramine maleate by micellar electrokinetic chromatography. AB - A micellar electrokinetic chromatography (MEKC) method was established for determination of paracetamol (PARA) and chlorpheniramine maleate (CPM) in cold tablets. Separation of both drugs, as well as other seven cold remedy ingredients, was achieved in 25.5 min using a sodium dihydrogenphosphate-sodium tetraborate buffer (10 mM, pH 9.0) containing sodium dodecyl sulfate (SDS) (50 mM) and acetonitrile (26% v/v). The effective capillary length of 50 cm, the separating voltage of 15 kV and the temperature of 30 degrees C was optimized. Detection was by a diode array detector at 214 nm. Method linearity was excellent (r(2)>0.999) over the concentration tested (10-250 microg/ml) with good precision and accuracy. Recoveries were good (>99%) with limits of detection of 0.4 and 0.5 microg/ml and limits of quantitation of 2 (%R.S.D.=3.1%) and 4 (%R.S.D.=2.4%) microg/ml, for PARA and CPM, respectively. The developed method was applied to the determination of ingredients in cold tablets and was found to be simple, rapid and efficient. PMID- 14550864 TI - On-line SPE-CE for the determination of insulin derivatives in biological fluids. AB - An on-line SPE-CE system is described for the determination of insulin derivatives in urine, serum and plasma. By combining techniques based on different separation mechanisms, in this case reversed-phase SPE and CE, a more selective sample clean-up is obtained. The described on-line SPE-CE procedure is able to desalt and clean biological samples, resulting in more repeatable electrophoretic results as well as a good linearity for urine, serum and plasma samples spiked with insulin derivatives, thus proving the elimination of detrimental effects caused by the sample matrix. The on-line SPE-CE system was linear for urine, serum and plasma samples spiked with insulin derivatives between 5 and 80 mg/l. The repeatability in migration time was below 1% relative standard deviation (R.S.D.). The repeatability of the peak was better (<2.4% R.S.D.) when no off-line precipitation reaction (<6.2% R.S.D.) was used, proving the beneficial characteristics of on-line sample pretreatment procedures over off line sample pretreatment procedures which are prone to sample losses and contamination. PMID- 14550865 TI - Determination of heroin metabolites in human urine using capillary zone electrophoresis with beta-cyclodextrin and UV detection. AB - A method has been developed for the detection of a mixture of morphine, codeine, 6-acetyl morphine (6-AM) and normorphine using capillary zone electrophoresis (CZE). The method utilized urinary 6-AM as a diagnostic indicator of heroin abuse because it is not a product of either morphine or codeine metabolism. The electrophoretic separation was achieved using an uncoated (50 microm I.D.) fused silica capillary, 77 cm long, containing the detector window 10.0 cm from the outlet end. The running buffer (pH 6.0) contained 50 mM sodium phosphate and 0.015 M beta-cyclodextrins (beta-CD). The samples were first extracted using a mixed-mode solid-phase extraction procedure and then analyzed by CZE. The UV absorbance detection was monitored at 214 nm. It has been found that beta-CDs can improve separation efficiency due to their hydrophobic cavity. The effect of the concentration of beta-CD and pH was also evaluated. The application of electrokinetic injection with field amplified sample stacking results in low detection limits (40 ng/ml for each analyte) and the method has good reproducibility, precision, accuracy, and high recovery. PMID- 14550866 TI - A validated assay for measuring doxorubicin in biological fluids and tissues in an isolated lung perfusion model: matrix effect and heparin interference strongly influence doxorubicin measurements. AB - Doxorubicin is an antineoplasic agent active against sarcoma pulmonary metastasis, but its clinical use is hampered by its myelotoxicity and its cumulative cardiotoxicity, when administered systemically. This limitation may be circumvented using the isolated lung perfusion (ILP) approach, wherein a therapeutic agent is infused locoregionally after vascular isolation of the lung. The influence of the mode of infusion (anterograde (AG): through the pulmonary artery (PA); retrograde (RG): through the pulmonary vein (PV)) on doxorubicin pharmacokinetics and lung distribution was unknown. Therefore, a simple, rapid and sensitive high-performance liquid chromatography method has been developed to quantify doxorubicin in four different biological matrices (infusion effluent, serum, tissues with low or high levels of doxorubicin). The related compound daunorubicin was used as internal standard (I.S.). Following a single-step protein precipitation of 500 microl samples with 250 microl acetone and 50 microl zinc sulfate 70% aqueous solution, the obtained supernatant was evaporated to dryness at 60 degrees C for exactly 45 min under a stream of nitrogen and the solid residue was solubilized in 200 microl of purified water. A 100 microl volume was subjected to HPLC analysis onto a Nucleosil 100-5 microm C18 AB column equipped with a guard column (Nucleosil 100-5 microm C(6)H(5) (phenyl) end capped) using a gradient elution of acetonitrile and 1-heptanesulfonic acid 0.2% pH 4: 15/85 at 0 min-->50/50 at 20 min-->100/0 at 22 min-->15/85 at 24 min- >15/85 at 26 min, delivered at 1 ml/min. The analytes were detected by fluorescence detection with excitation and emission wavelength set at 480 and 550 nm, respectively. The calibration curves were linear over the range of 2-1000 ng/ml for effluent and plasma matrices, and 0.1 microg/g-750 microg/g for tissues matrices. The method is precise with inter-day and intra-day relative standard deviation within 0.5 and 6.7% and accurate with inter-day and intra-day deviations between -5.4 and +7.7%. The in vitro stability in all matrices and in processed samples has been studied at -80 degrees C for 1 month, and at 4 degrees C for 48 h, respectively. During initial studies, heparin used as anticoagulant was found to profoundly influence the measurements of doxorubicin in effluents collected from animals under ILP. Moreover, the strong matrix effect observed with tissues samples indicate that it is mandatory to prepare doxorubicin calibration standard samples in biological matrices which would reflect at best the composition of samples to be analyzed. This method was successfully applied in animal studies for the analysis of effluent, serum and tissue samples collected from pigs and rats undergoing ILP. PMID- 14550867 TI - Experiences with monolithic LC phases in quantitative bioanalysis. AB - The applicability of monolithic liquid chromatographic (LC) phases in the field of quantitative bioanalysis has been evaluated. Two existing methods with fluorescence detection (the determination of bexarotene in plasma and the determination of dextromethorphan plus metabolites in urine) were successfully transferred from a conventional reversed-phase column to a 10 cm x 4.6 mm i.d. monolith. By simply increasing the mobile phase flow-rate, run times were about 3 fold reduced, while the chromatographic resolution of the analytes remained unaffected. In both cases, a very good correlation was found between the results of clinical samples obtained with the original method and those obtained with the adapted method. Two methods with tandem mass spectrometric detection were set up. For one of these methods (nifedipine in plasma), the separation of the analyte from interfering matrix components did not need a high plate number; the resolution found on a 10-cm monolith at 6 ml/min and that on a 3-cm conventional column at 2 ml/min were comparable and achieved in the same period of time. As the validation results on both column types were similar and considering the limited compatibility of mass spectrometric detection with high solvent flow rates, the conventional column was concluded to be the best choice for this application. For the determination of estradiol in plasma, however, there was so much interfering material that needed to be separated from the analyte, that the best results were obtained with three 10-cm monolithic columns coupled in series and because of the possibility to apply a relatively high flow-rate, a reasonable run time was still achieved. PMID- 14550868 TI - Simultaneous determination of moexipril hydrochloride and hydrochlorothiazide in tablets by derivative spectrophotometric and high-performance liquid chromatographic methods. AB - Two new simple and selective assay methods have been presented for the binary mixtures of moexipril hydrochloride (MOEX) and hydrochlorothiazide (HCTZ) in pharmaceutical formulations. The first method depends on second-derivative ultraviolet spectrophotometry with zero-crossing measurements at 215 and 234 nm for MOEX and HCTZ, respectively. The assay was linear over the concentration ranges 1.0-11.0 microg ml(-1) for MOEX and 0.5-9.0 microg ml(-1) for HCTZ. The determination limits for MOEX and HCTZ were found to be 1.0 and 0.5 microg ml( 1), respectively; while the detection limits were 0.2 microg ml(-1) for MOEX and 0.1 microg ml(-1) for HCTZ. The second method was based on isocratic reversed phase liquid chromatography by using a mobile phase acetonitrile-20 mM phosphate buffer (pH 4.0) (50:50, v/v). Lisinopril was used as an internal standard (IS) and the substances were detected at 212 nm. The linearity range for both drugs was 0.5-12.0 microg ml(-1). The determination and detection limits were found to be 0.100 and 0.010 microg ml(-1) for MOEX and 0.025 and 0.005 microg ml(-1) for HCTZ, respectively. The proposed methods were successfully applied to the determination of these drugs in synthetic mixtures and commercially available tablets with a high percentage recovery, good accuracy and precision. PMID- 14550869 TI - High performance liquid chromatographic determination, pharmacokinetic and comparative bioavailability studies of cisapride. AB - A sensitive and specific reversed phase HPLC method was developed to quantitate plasma levels of cisapride in order to conduct comparative bioavailability studies. The drug and internal standard was extracted from plasma with heptane isoamyl alcohol (95:5 v/v) and back extracted with sulfuric acid. The acidic layer was then re-extracted with the same extracting solvent. The separated organic layer was evaporated to dryness under nitrogen and the residue reconstituted with acetonitrile. Analysis was performed on a C-8 Sil-X-10 HPLC column, with a mobile phase of acetonitrile, water, and triethylamine (75:25:0.01) and UV detection at 215 nm. The standard curve covering the concentration range 5-160 ng/ml was linear (r(2)=0.9992), relative errors were within +/-10% and the CV% ranged from 1.34 to 11.82. The in vivo study was carried out in 12 healthy volunteers according to a single dose, two-sequence, cross over randomized design. The bioavailability was compared using the total area under the plasma level versus time curve (AUC(0-34,) AUC(0- infinity )), peak plasma concentration (C(max)) and time to C(max) (T(max)). No statistically significant difference was found between the AUC(0- infinity ) or C(max) values of the test (cisapride) and reference (Propulsid). It was, therefore, concluded that the generic cisapride was bioequivalent with the innovator formulation. PMID- 14550871 TI - Primary sclerosing cholangitis. AB - Primary sclerosing cholangitis (PSC) is a fibrosing disease of the intra- and extra-hepatic bile ducts, and is closely associated with inflammatory bowel disease. It is immune mediated, rather than being a classical autoimmune disease. A range of immune abnormalities have been demonstrated in PSC, in particular the findings of a range of autoantibodies, a portal tract infiltrate of functional T cells, a restricted T-cell receptor repertoire, and aberrant expression of HLA molecules on biliary epithelial cells. The immunogenetics of PSC is currently under study and to date 4 key HLA haplotypes associated with PSC have been developed. The trigger factor for the initiation of the immune response may be the ingress of bacteria or other toxic metabolites into the portal circulation through a diseased and permeable bowel wall. PMID- 14550870 TI - SPE-HPLC method for the determination and pharmacokinetic studies on paeoniflorin in rat serum after oral administration of traditional Chinese medicinal preparation Guan-Xin-Er-Hao decoction. AB - A new HPLC method for the determination of paeoniflorin in rat serum with solid phase extraction (SPE) for preconcentration is introduced. Paeoniflorin and an internal standard (pentoxifylline) were extracted from serum by means of SPE using cartridges with octadecyl chemically bound phase. The HPLC separation was then performed on a reversed-phase C(18) column using acetonitrile-water (18:82, v/v) as eluting solvent system, and UV detection at 230 nm to measure the analyte with a limit of quantitation about 10 ng ml(-1). The calibration curve for paeoniflorin was linear (r=0.9938) in the concentration range of 10-1200 ng ml( 1), both intra- and inter-day precision of the paeoniflorin were determined and their coefficience of variation did not exceed 10%. The validated method has been successfully applied for pharmacokinetic studies of paeoniflorin from rat serum after oral administration of Guan-Xin-Er-Hao decoction. PMID- 14550872 TI - Autoantigenicity of nucleolar complexes. AB - Autoantibodies targeting nucleolar autoantigens (ANoA) are most frequently found in sera from patients with systemic sclerosis (SSc, also designated scleroderma) or with SSc overlap syndromes. During the last decade an extensive number of nucleolar components have been identified and this allowed a more detailed analysis of the identity of nucleolar autoantigens. This review intends to give an overview of the molecular composition of the major (families of) autoantigenic nucleolar complexes, to provide some insight into their functions and to summarise the data concerning their autoantigenicity. PMID- 14550873 TI - Autoimmunity and hepatitis C. AB - Hepatitis C is a widespread chronic liver disease leading to cirrhosis and to the complications of portal hypertension. Based on biochemical and clinical features, it is almost indistinguishable from autoimmune hepatitis, which is characterized by the absence of viral infection, and other causes of chronic liver diseases, and represents a classical autoimmune disease with loss of immunological tolerance of liver tissue. Although the differentiation between both diseases is not difficult due the availability of diagnostic viral markers, it is well recognized that not only are autoantibodies present in autoimmune hepatitis frequently detected in hepatitis C, but also that an array of immune-mediated symptoms and diseases occur in patients with chronic hepatitis C. This has prompted research aimed at identifying a link between hepatitis C and autoimmunity, and autoimmune hepatitis in particular. This review focuses on the general immunological mechanisms linking viral infections with autoimmunity and includes the specific features of hepatitis C- and D-associated autoimmunity. Virus infection remains at the center of molecular and cellular research aimed at identifying the forces driving human autoimmunity and autoimmune diseases. PMID- 14550874 TI - Atherosclerosis: anti-inflammatory and immunomodulatory activities of statins. AB - Large clinical trials have demonstrated that HMG-CoA reductase inhibitors or 'statins' greatly reduce cardiovascular-related morbidity and mortality. The beneficial effects of statins were first assumed to result from their ability to reduce cholesterol synthesis and improve serum lipid profiles. In recent years, however, numerous pleiotropic effects of statins on atherosclerotic lesions have been described. In this review, we summarize the actions of statins on different aspects of atherosclerotic plaque development. These include endothelial dysfunction, leukocyte recruitment and inflammation, smooth muscle cell activation/proliferation, and finally plaque rupture and thrombosis. PMID- 14550875 TI - Autoimmunity seen through the SEREX-scope. AB - Autoantibodies can be detected in autoimmune diseases with a long prodromal phase and may serve as early indicators of disease activity. Autoantibody-based screening methods are therefore potent tools for the identification of target antigens. The SEREX method (serological identification of antigens by recombinant expression cloning) has been developed for the serological definition of immunogenic tumor antigens. Recent studies indicate that the SEREX approach may also be utilized for the analysis of complex immune responses involved in autoimmune diseases. PMID- 14550876 TI - The biology of TNF blockade. AB - Rheumatoid arthritis and Crohn's disease are costly diseases that result in significant long-term patient disability. They are chronic inflammatory diseases that are associated with increased production of Tumor Necrosis Factor (TNF). Blockage of this cytokine with bio-engineered compounds has significantly changed therapy of these diseases and has ushered in the era of biological therapy. The pro-inflammatory role of TNF is mediated by its essential respiratory burst function that is effectively inhibited by anti-TNF therapy. Anti-TNF therapy is effective in approximately two-thirds of patients to whom it is administered, but the effect is temporary. Lack of response to anti-TNF therapy stems from interplay of host-factors including: host cytokine response, disease phenotype, and antibody response to the anti-TNF agents. NOD 2, a defect present in approximately 50% of Crohn's disease patients, bears no relationship to non response. Additionally, TNF promoter gene polymorphisms and TNF receptor gene heterogeneity play a significant role in non-response and disease course/severity. Adverse effects of anti-TNF therapy include early and delayed hypersensitivity reactions, cell-mediated infections, lupus-like syndrome, demyelinating diseases, and exacerbation of CHF. PMID- 14550877 TI - Association of celiac disease with connective tissue diseases and autoimmune diseases of the digestive tract. AB - Celiac disease (CD) is an autoimmune enteropathy triggered by the ingestion of gluten in susceptible individuals, and is one of the most frequent genetically based diseases, with a prevalence of 1:200 in the general population. The association between CD and connective tissue diseases (CTD) and autoimmune diseases of the digestive tract (DT) has been described in several case reports but in few extensive studies, with varying prevalence. A high rate of false positive results were observed when low specific tests, such as the anti-gliadin and the guinea pig tissue transglutaminase (tTG) antibody assays were used. In a study of 400 patients with CTD and 218 with autoimmune DT disease, tested for IgA and IgG anti-tTG using the more specific human recombinant antigen, 12 cases (1.9%) of anti-tTG antibody positivity were found, but only 2 (0.3%) were confirmed as affected by CD following small bowel biopsy. Most of the patients testing false positive had primary biliary cirrhosis. In this short review we describe the association between CD and CTD, inflammatory bowel disease and primary biliary cirrhosis, with special emphasis on the diagnostic accuracy of CD antibody assays. PMID- 14550878 TI - Immune deficiency and autoimmunity. AB - Immunodeficiency and autoimmune phenomena may occur concomitantly in the same individual. Many immune deficiency syndromes, mainly humoral defects, are associated with autoimmune disorders. Hematological manifestations, such as thrombocytopenia and hemolytic anemia, are the most common presentation, but many other autoimmune mediated conditions have also been described. Persistent antigen stimulation, due to an inherently defective immune system ability to eradicate pathogenesis is the primary cause leading to autoimmunity in patients with primary immunodeficiency states. Other factors leading to the increase incidence of autoimmune manifestion will be discussed in the present review. Treatment with intravenous gammagluobuilin may ameliorate the autoimmune disorder and bone marrow transplantation can cure both conditions. PMID- 14550879 TI - The assessment of the patient with systemic sclerosis. AB - Systemic sclerosis presents a great deal of variability among different patients in the extent of skin and internal organ involvement, the pace of the disease and consequently, the prognosis. In addition, the single patient, during his/her disease course, can present with distinct manifestations. Each patient must, therefore, be carefully investigated. The assessment should be adapted to the setting, i.e. clinical practice, clinical investigation, therapeutic trials. The clinician cannot confine the diagnostic process to labelling the disease. He must define the subset, the extent of internal organ involvement, and the type of lesions underlying the clinical manifestations, i.e. fibrotic lesions, reflecting irreversible damage should be separated from active lesions (such as alveolitis) which can be reversed by drug treatment. The clinical investigator must assure that his/her patients are comparable to other series. ACR criteria have been shown to not assure such comparability. To this purpose, a core set of variables to be assessed in any clinical investigation study has been proposed. Finally, the clinical researcher should enrol patients with active disease and must rely for his/her conclusions on feasible and sensitive to change measures. An OMERACT committee has recently reviewed the literature selecting those ready for use in clinical trials. PMID- 14550880 TI - The etiology of autoimmune diabetes and thyroiditis: evidence for common genetic susceptibility. AB - Type 1 diabetes (T1D) and the autoimmune thyroid diseases (AITD) are the most common autoimmune endocrine diseases. Both are organ-specific T-cell mediated diseases. Abundant epidemiological data support a strong genetic basis for both T1D and AITD. Furthermore, both diseases commonly occur in the same individuals and in the same families. Indeed, studies suggest that the etiology of T1D and AITD may involve common genetic factors. Two immune regulatory genes, HLA and CTLA-4 contribute to the susceptibility to both diseases. Additionally, two tissue-specific genes, the insulin VNTR in T1D and Thyroglobulin in AITD play a major role in their pathogenesis. Therefore, it is likely that both immune regulatory and target tissue genes contribute to these and other autoimmune diseases. PMID- 14550881 TI - HLA class II alleles and genetic predisposition to the antiphospholipid syndrome. AB - The antiphospholipid syndrome (APS) is an autoimmune disease characterized by the presence of antiphospholipid antibodies (aPL). Its etiology is linked to genetic predisposition, which is accounted for, at least in part, by genes of major histocompatibility complex (HLA system). The association of APS with human leukocyte antigen (HLA) alleles is a consequence of the association of aPL with HLA alleles. Some HLA alleles carry the risk to produce aPL, and this is independent of the clinical context. In fact, we find the same associations between HLA and aPL in primary APS and in APS secondary to systemic lupus erythematosus (SLE). The association of HLA-DR4, -DR7, -DRw53 and -DQB1*0302 with aCL that has been demonstrated in primary APS can also be found in SLE, a disease with a completely different pattern of HLA allele association (DR2, DR3, DRw52). In addition, the various aPL (anticardiolipin antibodies, lupus anticoagulant, anti-beta2GPI antibodies, antiphosphatidylserine/prothrombin antibodies) show similar HLA association, again independent of the clinical context (primary APS or SLE), and across various ethnic groups. PMID- 14550882 TI - Introduction to micro-analytical systems: bioanalytical and pharmaceutical applications. AB - This review presents a brief overview of recent developments in miniaturization of analytical instruments utilizing microfabrication technology. The concept 'Micro-Total Analysis Systems micro-TAS)', also termed 'Lab-on-a-chip', and the latest progresses in the development of microfabricated separation devices and on chip detection techniques are discussed. Applications of micro-analytical methods to bioanalytical and pharmaceutical studies are also described, including chemical reactions, assays, and analytical separations of biomolecules in micro scale. PMID- 14550883 TI - Convergent synthesis and antibacterial activity of pyrazole and pyrazoline derivatives of diazepam. AB - Polysubstituted pyrazoles (5)(a-l), pyrazolines (7)(a-c), (8)(a-c) and pyrazolotriazine (10) derivatives of diazepam were synthesized. The structures of hitherto unknown compounds were established by analytical and spectral methods. Some of these compounds were screened to test their antibacterial activity against gram-positive (B. subtilis) and gram-negative (P. aeruginosa). All compounds showed potent activity against these bacteria. PMID- 14550884 TI - Permeation, metabolism and site of action concentration of nicotinic acid derivatives in human skin. Correlation with topical pharmacological effect. AB - A novel methodology for establishing a pharmacological dose-effect relationship of methyl nicotinate, hexyl nicotinate and nicotinic acid acting as peripheral vasodilators in the skin following topical application is investigated. This methodology involves the estimation of the unbound drug concentration in the aqueous compartment at the site of action in tissue, termed C(*), which was evaluated as the pertinent concentration responsible for the pharmacological effect. Blood capillaries next to the epidermis-dermis boundary were postulated to be the relevant site of action. C(*) was estimated from drug transport parameters for different layers of human cadaver skin determined in vitro. Immunohistochemical studies showed that the plane of separation of skin achieved by heat treatment was between the basal cells of the epidermis and the lamina lucida, confirming the integrity of the epidermis and the dermis used in the experiments. The permeation rate for epidermis increased drastically with increasing lipophilicity of the drug. Dermis permeability was roughly the same for all three compounds. The epidermis represented the major transport barrier in vitro for methyl nicotinate and nicotinic acid but not for hexyl nicotinate. The esters were metabolised to nicotinic acid during tissue permeation to an extent that was rather limited for the epidermis but very pronounced for the dermis. Nonspecific alpha-naphthylacetate-esterase activity was predominantly located in the dermis, which was in agreement with the metabolism results. The drugs were applied each at three different concentrations in vivo to the ventral forearm of healthy human volunteers and vasodilation was evaluated based on skin erythema which was quantified by measuring colour change of reflected light. Area under the curve of the change of colour co-ordinates as a function of time was used as a measure of pharmacological effect. The pharmacological effect of all three drugs was comparable when similar C(*) values were considered, even though the concentrations applied to the skin differed by orders of magnitude. The effect showed a strong positive dependence on C(*). Methyl and hexyl nicotinate showed identical, nearly sigmoidal effect/C(*)-profiles, while the profile for nicotinic acid was linear, suggesting a possible difference in the intrinsic pharmacological potency between the esters and the acid. These results demonstrate the validity of C(*) as the relevant drug concentration for the cutaneous pharmacological effect of the topically applied drugs and underline the usefulness of the presented methodology for establishing dose-response relationships in dermal therapy and expressing bioavailability. PMID- 14550885 TI - Driving forces and the influence of the buffer composition on the complexation reaction between ibuprofen and HPCD. AB - Cyclodextrins are often used in order to increase the aqueous solubility of drug substances by complexation. In order to investigate the complexation reaction of ibuprofen and hydroxypropyl-beta-cyclodextrin, titration calorimetry was used as a direct method. The thermodynamic parameters of the complexation process (stability constant, K(11); complexation enthalpy, deltaH(c) degrees ) were obtained in two different buffer systems (citric acid/sodium-phosphate and phosphoric acid) at various pH values. Based on these data the relative contributions of the enthalpic and entropic terms of the Gibbs energy to the complexation process have been analyzed. In both buffers the enthalpic and entropic terms are of different sign and this case corresponds to a 'nonclassical' model of hydrophobic interaction. In citric buffer, the main driving force of complexation is the entropy, which increases from 60 to 67% while the pH of the solution increases from 3.2 to 8.0. However, for the phosphoric buffer the entropic term decreases from 60 to 45%, while the pH-value of the solution increases from 5.0 to 8.2, and the driving force of the complexation process changes from entropy to enthalpy. The experimental data of the present study are compared to results of other authors and discrepancies discussed in detail. PMID- 14550887 TI - Rapid in vitro test to predict ocular tissue permeability based on biopartitioning micellar chromatography. AB - The drug permeability prediction across the ocular tissues is important in the development of new drugs and drug delivery strategies. Physicochemical characteristics of drugs, mainly acid-base character, hydrophobicity and the molecular size determine both their transport across the eye tissue barriers and their retention in biopartitioning micellar chromatography (BMC). An in vitro model able to describe and predict the whole cornea drug permeability is proposed. The model uses the retention of drugs in BMC and molecular weight (MW) as predictive variables. The relationships between drug retention data in BMC and their bibliographic permeability values in stroma, epithelium-plus-stroma and endothelium-plus-stroma are also studied. The results show that BMC can be a useful tool to select drug candidates according to their whole cornea permeability at the early stage of the drug discovery process. PMID- 14550886 TI - 2-(4,5-dihydro-1H-imidazol-2-yl)indazole (indazim) derivatives as selective I(2) imidazoline receptor ligands. AB - A series of variously substituted 2-(4,5-dihydro-1H-imidazol-2-yl)indazoles 3a-j and 2-(4,5-dihydro-1H-imidazol-2-yl)-4,5,6,7-tetrahydroindazole 6 were prepared by the regiospecific heteroalkylation of corresponding indazoles 1a-k with 2 chloro-4,5-dihydroimidazole (2). Their affinity to imidazoline I(2) receptors and alpha(2)-adrenergic receptors was determined by radioligand binding assay carried out on P(2) membrane preparations obtained from rat whole brains. 4-Chloro-2-(4,5 dihydro-1H-imidazol-2-yl)indazole (3f, 4-Cl-indazim) showed a 3076-fold difference in affinity for the [(3)H]2BFI-labeled imidazoline I(2) receptors relative to the [(3)H]RX821001-labeled alpha(2)-adrenergic receptors. This highly selective compound should prove to be useful tool in further understanding the functions of the imidazoline I(2) receptors. PMID- 14550888 TI - Synthesis of a novel fluorescent poly(D,L-lactide) end-capped with 1 pyrenebutanol used for the preparation of nanoparticles. AB - A new fluorescent polymer based on D,L-lactic acid units end-capped with 1 pyrenebutanol (PLAP) was synthesized by ring-opening polymerization. PLAP having different molecular weight could be obtained by varying the ratio of D,L-lactide and 1-pyrenebutanol. Fluorescent nanoparticles (NP) were prepared using blends of poly(D,L-lactic acid) (PLA) and the new PLAP of 6 kDa by the salting-out process. Incubation of these nanoparticles with human blood monocytes was performed in serum and the cell-associated fluorescence was analysed by flow cytometry. Monocytes in contact with NP containing increasing amounts of PLAP showed a regular increase of the fluorescence. Cells incubated with NP containing 5% (w/w) of PLAP showed high signals of fluorescence with no possible overlap with those given by blank monocytes. This demonstrated that flow cytometry performed in the UV domain was very specific. In addition, the results of cytotoxicity tests using a MTT assay method indicated that PLAP did not increase the cytotoxicity when incorporated into PLA nanoparticles. PMID- 14550889 TI - The effects of dose staggering on metabolic drug-drug interactions. AB - PURPOSE: To investigate the effect of dose staggering on metabolic drug-drug interactions (MDDI). METHODS: Using Matlab, anatomical, physiological and biochemical data relating to human pharmacokinetics were integrated to create a representative virtual healthy subject relevant to in vivo studies. The effects of dose staggering on AUC and C(max) were investigated under various scenarios with respect to pharmacokinetic characteristics of the inhibitor and substrate drugs (e.g. hepatic extraction ratio). Specific cases were also simulated where MDDI had been studied experimentally for combinations of drugs (budesonide and ketoconazole; triazolam and itraconazole). RESULTS: The decrease in the magnitude of the inhibitory effect of the 'perpetrator' drug (inhibitor) on the 'victim' drug (substrate) as a result of 'dose staggering' was greater when the 'perpetrator' was given after the 'victim'. There was reasonable agreement between the predicted extent of the interactions and the observed in vivo data (mean prediction errors of 25 and -14% for AUC and C(max) values, respectively (n=7)). The impact of dose staggering was minimal during continuous dosage of inhibitors with long elimination half-lives (e.g. itraconazole, >20 h). CONCLUSIONS: Clinical trial simulations using physiological information may provide useful guidelines for optimal dose staggering when poly-pharmacy is inevitable. PMID- 14550890 TI - Kinetics of degradation of 4-imidazolidinone prodrug types obtained from reacting prilocaine with formaldehyde and acetaldehyde. AB - The kinetics of decomposition of 4-imidazolidinone prodrug types obtained by reacting prilocaine (I) with formaldehyde and acetaldehyde has been studied in aqueous solution in the pH range 1-7.4 at 60 and 37 degrees C, respectively. At pH<5 the hydrolysis of the derivative derived from formaldehyde (II) to yield I obeyed apparent first-order kinetics. At higher pH, the decomposition reactions proceeded to an equilibrium and the reactions could be described by first- and second-order reversible kinetics. A plot of the logarithm of the apparent first order rate constants for hydrolysis of II against pH resulted in a sigmoidal shaped pH-rate profile characteristic for the hydrolysis of many N-Mannich bases. A half-life at pH 7.4 (60 degrees C) of 6.9h for compound II was calculated. Compared to II the 4-imidazolidinone derived from acetaldehyde (III) exhibited enhanced instability in aqueous buffer solutions. The decomposition was followed at 37 degrees C monitoring the decrease in concentration of intact (III). At acidic pH the reactions displayed strict first-order kinetics and the disappearance of III was accompanied by a concomitant formation of I. At pH 7.4, the rate data also applied reasonably well to first-order kinetics despite the observation that small amounts of III was formed at pH 7.4 from a solution containing equimolar concentrations of acetaldehyde and prilocaine (10(-4)M). In case of III, a bell-shaped pH-rate profile was obtained by plotting the logarithm of the pseudo-first-order rate constants against pH indicating the involvement of a kinetically significant intermediate in the reaction pathway and a change of the rate-limiting step in the overall reaction with pH. For the stability studies performed at pH 6.9 and 7.4 product analysis revealed that parallel to formation of (I) an unknown compound (X) emerged. Compared to III, compound X is hydrolysed to give I at a slower rate (t(50%)=30 h at 37 degrees C). Based on LC-MS data it is suggested that (X) is an isomeric form of III, which may exist in four diastereomeric forms. Thus, at physiological pH an initial relatively fast regeneration of I from III is to be expected followed by a slower drug activation resulting from hydrolysis of the isomeric form of III. PMID- 14550891 TI - Plasmepsin 4, the food vacuole aspartic proteinase found in all Plasmodium spp. infecting man. AB - Plasmepsins are aspartic proteinases of the malaria parasite, and seven groups of plasmepsins have been identified by comparing genomic sequence data available for the genes encoding these enzymes from Plasmodium falciparum, Plasmodium vivax, Plasmodium knowlesi, Plasmodium berghei, and Plasmodium yoelii. The food vacuole plasmepsins typified by plasmepsin 4 from P. falciparum (PfPM4) constitute one of these groups. Genes encoding the ortholog of PfPM4 have been cloned from Plasmodium ovale, Plasmodium malariae, and P. vivax. In addition, P. falciparum contains three paralagous food vacuole plasmepsins or plasmepsin-like enzymes that appear to have arisen by gene duplication, plasmepsins 1 (PfPM1), 2 (PfPM2) and HAP, and all four were localized to purified food vacuole preparations by two dimensional gel electrophoresis and mass spectroscopic analysis. The three paralogs of PfPM4 do not have counterparts in the six other Plasmodium spp. examined by genomic DNA blot analysis and by review of available genomic sequence data. The presence of these paralogs among the food vacuole plasmepsins in P. falciparum as compared with the other three species causing malaria in man will impact efforts to rationally design antimalarials targeting the food vacuole plasmepsins. PMID- 14550892 TI - Identification of genotypically mixed Cryptosporidium parvum populations in humans and calves. AB - Genotypic analyses of Cryptosporidium parvum oocysts have divided the species into two genotypes, referred to as type 1 and type 2. Although humans are susceptible to both types, mixed type 1/type 2 infections have rarely been identified. The paucity of mixed infections could be explained by the predominance of one type over the other in mixed infections, or by the poor sensitivity of restriction fragment length polymorphism (RFLP) analyses for detecting subpopulations. Using a type-specific real-time PCR assay capable of detecting type 1 or type 2 constituting as little as 0.01% of the population, archived and new isolates of human, bovine, and mouse origin were genotyped. Mixed type 1/type 2 infections were identified in humans and calves, including in samples previously found to be homogeneous by RFLP. Isopycnic fractionation of mixed isolates revealed that type 1 and type 2 oocysts differ in their sedimentation properties. The detection of a type 1 subpopulation in serially propagated bovine isolates indicates that type 1 and type 2 are stably maintained during long-term passage. Together with recently reported experimental bovine and ovine type 1 infections, the persistence of type 1 subpopulation in experimentally infected animals suggests that animals may play a previously unrecognized role in the maintenance of C. parvum type 1. PMID- 14550893 TI - Specific cleavage sites on human IgG subclasses by cruzipain, the major cysteine proteinase from Trypanosoma cruzi. AB - Cruzipain, the major cysteine proteinase of Trypanosoma cruzi, might have other biological roles than its metabolic functions. In this report, we have explored the interaction of cruzipain with molecules of the immune system. The enzyme was used to digest all human IgG subclasses at different pH values and lengths of time. At pH 7.3, all subclasses were readily split at the hinge region. Immunoblot and amino acid sequence analysis showed fragments of IgG1 and IgG3 to be compatible with Fab and Fc, whereas IgG2 and IgG4 rendered Fab2 and Fc. In all cases the fragments produced might impair the binding capacities and the effector functions of specific IgG. At these cleavage sites cruzipain displays cathepsin L and/or cathepsin B activities and shows a clear preference for Pro at the P'2 position and polar residues at P1. Despite the activity of cruzipain within the hinge, the enzyme also cleaved all heavy chains between the CH2 and CH3 domains; producing Fc'-like-fragments of 14 kDa. These fragments are potential candidates to block or saturate Fc receptors on immunocompetent cells. At mild acidic pH cruzipain produced further degradation of the Fc of all subclasses, the Fd of IgG4 and partially the Fd of IgG1, with the consistent loss of any antibody activity. The L chains apparently were not affected. Thus, cruzipain should be able to modulate, depending on the subclass selected and the pH of the environment, the production and the length of different biologically active/inactive IgG fragments. PMID- 14550894 TI - An in vitro system for developmental and genetic studies of Leishmania donovani phosphoglycans. AB - Glycoconjugates have been shown to play important roles in Leishmania development. However, the ability to study these molecules and other processes would benefit greatly from improved methods for genetic manipulation and analysis of the amastigote stage. This is especially challenging for L. donovani, the agent of the most severe form of leishmaniasis, which can rapidly lose virulence during in vitro culture. Here we report on a clonal subline of an L. donovani 1S2D (LdBob or LdB), which differentiates readily from promastigotes to amastigotes in axenic culture, and maintains this ability during extended parasite cultivation in vitro. This derivative can be plated and transfected efficiently while grown as promastigotes or amastigotes. Importantly, LdB maintains the ability to differentiate while undergoing genetic alterations required for creation of gene knockouts and complemented lines. Like virulent L. donovani, LdB exhibits down-regulation of lipophosphoglycan (LPG) synthesis and up-regulation of A2 protein synthesis in amastigotes. We showed that knockouts of LPG2, encoding a Golgi GDP-mannose transporter, eliminated phosphoglycan synthesis in LdB axenic amastigotes. These and other data suggest that LdB axenic amastigotes will be generally useful as a differentiation model in studies of gene expression, virulence, glycoconjugate function and drug susceptibility in L. donovani. PMID- 14550895 TI - Culex pipiens pipiens: characterization of immune peptides and the influence of immune activation on development of Wuchereria bancrofti. AB - Stimulating or augmenting the innate immune response of insect vectors has been shown to impede or disrupt the development and transmission of eukaryotic pathogens; however, the majority of such studies have utilized model systems and not natural parasite-vector systems. The Culex pipiens complex of mosquitoes functions as a primary urban vector of Wuchereria bancrofti, a causative agent of lymphatic filariasis. To test the effects of immune activation on this vector parasite interaction, Culex pipiens pipiens from the filariasis-endemic Nile Delta were subjected to bacteria inoculation and subsequently fed a blood meal containing W. bancrofti. No difference was seen between parasite development in these mosquitoes as compared to non-inoculated controls. A set of expressed sequence tags from blood-fed midgut and bacteria-inoculated Cx. p. pipiens reveals transcripts for the immune peptides cecropin, gambicin and defensin--all of which have been reported to have antiparasitic effects. Sequences and transcriptional profiles for these peptides are reported. The discrepancy between these results and those reported for the model parasite, Brugia malayi, in the mosquito Aedes aegypti are discussed. PMID- 14550896 TI - Gene structure and ookinete expression of the chitinase genes of Plasmodium vivax and Plasmodium yoelii. PMID- 14550897 TI - Haemonchus contortus UNC-18 binds to Caenorhabditis elegans neuronal syntaxin. PMID- 14550898 TI - Heterologous promoter activity in stable and transient Plasmodium knowlesi transgenes. PMID- 14550899 TI - Phenotypic diversity of smooth muscle cells isolated from human intracranial basilar artery. AB - The present work examined heterogeneity of vascular smooth muscle cells cultured from human cerebral arteries that has not been previously reported. Primary smooth muscle cell cultures were isolated from human intracranial basilar arteries. Using a ring isolation method, multiple clones were generated from the cell cultures. These clones had two distinctly different morphologies: (1) fusiform; and (2) stellate. At confluence the fusiform-shaped clones grew in compact clusters with overlapping cells while the stellate-shaped clones were contact-inhibited growing in a monolayered pattern. The smooth muscle differentiation markers, alpha-smooth muscle-actin, calponin and smooth muscle myosin heavy chains were expressed in all these clones. In response to serum stimulation, the stellate-shaped clones had a higher growth rate than the fusiform clones. This study reports that smooth muscle cells derived from human basilar arteries are heterogeneous. PMID- 14550900 TI - Dose ratio is important in maximizing naloxone enhancement of nalbuphine analgesia in humans. AB - The analgesic effect of kappa partial agonist opioids (i.e. nalbuphine, pentazocine and butorphanol) is significantly greater in women. Recent evidence suggests that this sexual dimorphism may result from a naloxone-sensitive anti analgesic effect that is activated along with, and summates with, the analgesic effect of these agents, resulting in decreased analgesia or increased pain. For example, nalbuphine (5 mg) produces profound anti-analgesia (i.e. enhanced pain) in men, but addition of a low dose of the opioid receptor antagonist naloxone (0.4 mg, opioid antagonist) induces significant analgesia in men and enhances nalbuphine analgesia in women. To further delineate the dose-dependent relationship of nalbuphine and naloxone, we recently evaluated the effect of a lower dose of nalbuphine (2.5 mg) with and without naloxone (0.4 mg) on dental postoperative pain. In women, nalbuphine alone induced modest short duration analgesia, which was antagonized by the addition of naloxone. In men, this dose of nalbuphine alone did not produce analgesia or anti-analgesia, and naloxone did not alter the response to nalbuphine. Thus, it appeared that the 2.5 mg dose of nalbuphine was not sufficient to induce anti-analgesia while the 0.4 mg dose of naloxone was able to antagonize the analgesic effect of nalbuphine, at least in women. In the current study, we tested the hypothesis that an important determinant of naloxone enhancement of nalbuphine analgesia is the dose ratio of nalbuphine to naloxone. Since a dose ratio of 12.5:1 (i.e. 5 mg nalbuphine:0.4 mg naloxone) resulted in analgesic enhancement, but a dose ratio of 6.25:1 (2.5 mg:0.4 mg) did not, we tested the same, lower, dose of nalbuphine (2.5 mg) in combination with a lower dose of naloxone (0.2 mg) to maintain the 12.5:1 dose ratio. This lower dose of naloxone significantly prolonged the analgesic effect of nalbuphine in both men and women, suggesting that the anti-analgesic effect of nalbuphine is present in both sexes at the 2.5 mg dose and that the dose ratio of nalbuphine to naloxone is an important determinant of the analgesic efficacy of this combination. PMID- 14550901 TI - Anti-kindling effect of slow repetitive transcranial magnetic stimulation in rats. AB - The cerebrospinal fluid (CSF) of animals exposed to electroconvulsive shock (ECS) has anticonvulsant properties when injected into naive animals. The present study investigated whether the CSF of humans exposed to 1 or 10 Hz repetitive transcranial magnetic stimulation (rTMS) has similar properties. Using a 4 day rat flurothyl kindling seizure model we found that the kindling rate was significantly decreased by intraventricular injection of CSF from depressed patients exposed to 1 Hz rTMS. The CSF from patients that underwent 10 Hz rTMS showed a trend toward an increased kindling rate. These results support the similarity of ECS and rTMS and suggest that 1 Hz and 10 Hz rTMS produce distinct physiologic changes. PMID- 14550902 TI - Excitatory effects of oxytocin and cholecystokinin on oxytocin neurones: differences between virgin, pregnant and lactating rats. AB - Extracellular recordings were made from supraoptic oxytocin (OT) neurones in rat brain slices to investigate the effect of reproductive state on their excitation by OT and cholecystokinin (CCK). Stable background activity was induced by local glutamate application, and OT neurones were identified by their continuous firing and opioid inhibition. The excitatory effect of OT (indicated by percentage neurones excited, peak increment in firing, and response magnitude) was maximal in lactating rats, intermediate in virgin, and lowest in pregnant animals. Excitation by CCK was high in both lactating and virgin rats but, as with OT tests, was minimal in the pregnant group. These adaptations in the responses to OT and CCK may help to ensure that the firing of OT neurones is appropriately regulated to meet changing demands for OT release. PMID- 14550903 TI - Neonatal enucleation reduces the proportion of callosal boutons forming multiple synaptic contacts in rat striate cortex. AB - Although bilateral enucleation at birth produces marked abnormalities in the overall distribution and topography of interhemispheric callosal connections in rat visual cortex, it is not known whether it also alters the morphology of callosal synapses. Here we report on the effect of neonatal enucleation on the proportion of callosal boutons making multiple postsynaptic contacts. Synapses were analyzed in adult rats after injections of the anterograde tracer biotinylated dextran amine into the opposite striate cortex. Results show that neonatal enucleation produces a significant reduction in the proportion of callosal boutons making multiple postsynaptic contacts. PMID- 14550904 TI - Bimodal circadian expression of serotonin N-acetyltransferase mRNA in the retina of rats under restricted feeding. AB - The mammalian retina has an endogenous oscillator that regulates circadian melatonin production. We examined this mechanism in the retina of rats under restricted feeding (RF) by investigating the expression rhythms of the mRNA for serotonin N-acetyltransferase (NAT), which is the rate limiting enzyme for melatonin synthesis. Northern blots showed that RF shifted the phase of circadian mRNA expression rhythms of a circadian clock gene rat period 2 and a clock controlled gene D-site binding protein. However, the NAT mRNA showed a bimodal rhythm pattern with one peak at midnight and the other at midday after RF, although the mRNA expression showed a single peak at midnight after ad libitum feeding. Our results suggest that NAT mRNA in the rat retina is regulated by at least two circadian oscillatory mechanisms: one is affected by feeding and the other can oscillate independent to the imposed feeding cycles. PMID- 14550905 TI - Histamine elicits neuronal excitatory response of red nucleus in the rat via H2 receptors in vitro. AB - Perfusing slices with histamine (1-100 microM) produced an excitatory response in rat rubral neurons (118/132, 89.4%). The histamine-induced excitation was not blocked by the low-Ca2+/high-Mg2+ medium (n=10), supporting a direct postsynaptic action of the amine. Histamine H2 receptor antagonist ranitidine effectively blocked the excitatory response of rubral neurons to histamine (n=26), but H1 receptor antagonist triprolidine did not (n=24). The excitatory effect of histamine could be mimicked by dimaprit, a highly selective H2 receptor agonist (n=24), and the dimaprit-elicited excitation of the rubral neurons could be blocked by ranitidine (n=16), but not by triprolidine (n=9). In addition, H1 receptor agonist 2-pyridylethylamine could not elicit any response in rubral neurons (n=12). These results indicate that histamine excites red nucleus neurons through H2 receptors and suggest that the histaminergic afferent fibers may play an important functional role in the sensorimotor integration through the red nucleus. PMID- 14550906 TI - Alpha-synuclein expression in HEK293 cells enhances the mitochondrial sensitivity to rotenone. AB - Mitochondrial dysfunction has been implicated in the aetiology of sporadic Parkinson's disease but its role in the disease mechanism is not clear. We have investigated the short term effect of G209A mutant or wild-type alpha-synuclein expression upon mitochondrial function using stable inducible cell models. Mitochondrial respiratory chain activities and membrane potential were normal suggesting that increased wild-type or mutant alpha-synuclein expression did not directly affect these parameters. However, both wild-type and mutant G209A alpha synuclein expression enhanced the fall in mitochondrial membrane potential induced by the complex I inhibitor rotenone. This suggests an indirect interaction between alpha-synuclein expression and mitochondrial function which could render the mitochondria more vulnerable to inhibition by potential endogenous or exogenous factors found in dopaminergic neurones. PMID- 14550907 TI - 'Thought'--control of functional electrical stimulation to restore hand grasp in a patient with tetraplegia. AB - The aim of the present study was to demonstrate the first time the non-invasive restoration of hand grasp function in a tetraplegic patient by electroencephalogram (EEG)-recording and functional electrical stimulation (FES) using surface electrodes. The patient was able to generate bursts of beta oscillations in the EEG by imagination of foot movement. These beta bursts were analyzed and classified by a brain-computer interface (BCI) and the output signal used to control a FES device. The patient was able to grasp a cylinder with the paralyzed hand. PMID- 14550908 TI - Distribution of mu receptors in the ventral respiratory group neurons; immunohistochemical and pharmacological studies in decerebrate cats. AB - Immunoreactivity for mu receptors was investigated in 21 bulbar respiratory neurons, individually identified by intracellular recording and labeling with neurobiotin. In 14 of these neurons, effects of iontophoresed morphine were examined. Morphine hyperpolarized the membrane and decreased spike discharges in 4/6 augmenting inspiratory (aug-I), 4/5 postinspiratory (post-I) and 3/3 augmenting expiratory (aug-E) neurons. It had no effect on two aug-I and one post I neurons. Strong immunoreactivity for mu receptor was detected in the soma and dendrites of 5/8 aug-I, 5/7 post-I and 6/6 aug-E neurons. In the remaining three aug-I and two post-I neurons that included cells unresponsive to morphine, weak immunoreactivity was detected only in the dendrites. These results demonstrated wide, but uneven, distribution of mu receptors in bulbar respiratory neurons and suggest their contribution to respiratory depression by opioids. PMID- 14550909 TI - Depolarization of PC12 cells induces neurite outgrowth and enhances nerve growth factor-induced neurite outgrowth in rats. AB - Synaptic plasticity is clearly controlled by synaptic activity and by neurotrophin-dependent signaling. We have previously hypothesized that synaptic activity modulates concomitant neurotrophin receptor signaling, thereby integrating the activity state of a synapse with the state of neurotrophic support available at the synapse. Herein we present evidence in support of this hypothesis. Using PC12 cells as a model of the presynaptic element, we show that depolarization increases TrkA tyrosine phosphorylation in response to nerve growth factor (NGF). Moreover, we show that depolarization alone is sufficient to induce the tyrosine phosphorylation of TrkA. These findings are functionally relevant, as evidenced by our observation that depolarization alone induces neurite outgrowth, and that depolarization dramatically enhances neurite outgrowth in response to NGF, especially in primed PC12 cells. We conclude that normal synaptic function may depend upon the integration of synaptic activity and activity-dependent neurotrophin release and signaling, and that these findings have potential relevance to neural repair. PMID- 14550910 TI - Integration of heard and seen speech: a factor in learning disabilities in children. AB - Normal-learning children (NL) and children with learning disabilities (LD) reported their perceptions of unisensory (auditory or visual), concordant audiovisual (e.g. visual /apa/ and auditory /apa/) and conflicting (e.g. visual /aka/ and auditory /apa/) speech stimuli in quiet and noise (0 dB and -12 dB signal-to-noise ratio, SNR). In normal populations, watching such conflicting combinations typically changes auditory percepts ('McGurk effect'). NL and LD children identified unisensory auditory and congruent audiovisual stimuli similarly in all conditions. Despite being less accurate identifying unisensory visual stimuli, LD children were more likely than NL children to report hearing only the visual component of incongruent audiovisual stimuli at -12 dB SNR. Furthermore, LD children with brainstem timing deficits demonstrated a distinctive pattern of audiovisual perception. The results suggest that the perception of simultaneous auditory and visual speech differs between NL and LD children, perhaps reflecting variations in neural processing underlying multisensory integration. PMID- 14550911 TI - Enhanced expression of transforming growth factor-beta isoforms in the neural tube of embryos derived from diabetic mice exposed to cyclophosphamide. AB - We analyzed the expression pattern of transforming growth factor-beta isoforms (TGF-beta1, TGF-beta2 and TGF-beta3) in the developing brain of embryos derived from the normal and diabetic mice exposed to cyclophosphamide (CP), a cytotoxic teratogen. The CP-treated diabetic embryos showed significantly more TGF-beta1 and TGF-beta2 immunoreactive cells in the regions of telencephalon and diencephalon in comparison to that of CP-treated non-diabetic embryos. Moreover, no cells expressing TGF-beta isoforms were detectable in the developing brain of normal and diabetic embryos. The mRNA expression levels of TGF-beta isoforms were found to be significantly increased in the developing brain of CP-treated diabetic embryos compared to that of CP-treated non-diabetic embryos as measured by quantitative real time reverse transcription-polymerase chain reaction. The enhanced expression levels of TGF-beta isoforms appear to be associated with the increased frequency of neural tube defects observed in the diabetic embryos exposed to CP. PMID- 14550912 TI - Behavioral reinforcement of long-term potentiation in rat dentate gyrus in vivo is protein synthesis-dependent. AB - A transient, protein synthesis-independent long-term potentiation (early-LTP, <4 h) can be reinforced into a maintained protein synthesis-dependent late-LTP (>4 h) by specific electrical stimulation of limbic structures (J. Neurosci. 21 (2001) 3697). Similarly, LTP-modulation can be obtained by behavioral stimuli with strong motivational content. However, the requirement of protein synthesis during behavioral reinforcement has not been shown so far. Thus, we have studied here this specific question using a behavioral reinforcement protocol, i.e. allowing water-deprived animals to drink 15 min after induction of early-LTP. This procedure transformed early-LTP into late-LTP. Anisomycin, a reversible protein synthesis inhibitor, abolished behavioral LTP-reinforcement. These results demonstrate that behavioral reinforcement depends on protein synthesis. PMID- 14550913 TI - Age-induced modifications of static postural control in humans. AB - We examined how young and older adults adapt their posture to static balance tasks of increasing difficulty. Participants stood barefoot on a force platform in normal quiet, Romberg-sharpened and one-legged stance. Center of pressure (CoP) variations, electromyographic (EMG) activity of ankle and hip muscles and kinematic data were recorded. Both groups increased postural sway as a result of narrowing the base of support. Greater CoP excursions, EMG activity and joint displacements were noted in old compared to younger adults. Older adults displayed increased hip movement accompanied by higher hip EMG activity, whereas no similar increase was noted in the younger group. It is concluded that older adults rely more on their hip muscles when responding to self induced perturbations introduced by increased task constraints during quiet standing. PMID- 14550914 TI - Effect of early thyroxine treatment on brain-derived neurotrophic factor mRNA expression and protein amount in the rat medial septum/diagonal band of Broca. AB - There is evidence that morphological alterations concerning deficiency or abundance of thyroid hormones (TH) may be mediated by brain-derived neurotrophic factor (BDNF). It has been demonstrated that the mRNA-expression of BDNF is increased after TH-treatment during the first postnatal weeks. After transient treatment of newborn rats with thyroxine mRNA expression, protein concentration and number and size of BDNF-immunopositive neurons were quantified in the medial septum/vertical diagonal Band of Broca (MS/vDB). The number and size of BDNF immunopositive neurons were estimated in young (P10) and adult (4 months). The amount of mRNA and protein are significantly increased in TH-treated rats at P10 compared to control animals. TH-treated animals showed a significant decrease of BDNF-immunopositive cell numbers in the adulthood. The results demonstrate a correlated increase of BDNF mRNA and protein in the septum at P10 which is an important stage of differentiation processes in the septohippocampal system. These results provide further evidence that BDNF is a possible candidate for the mediation the TH effects in the MS/vDB. PMID- 14550915 TI - Female rats show an increased sensibility to the forced swim test depressive-like stimulus in the hippocampus and frontal cortex 5-HT1A receptors. AB - Affective disorders are more common in women. The forced swim test acts like a depressive stimulus. Hippocampus and frontal cortex 5-HT1A receptors of female and male Wistar rats subjected to the forced swim test were compared with a sham group. The forced swim test diminishes (P<0.05) the hippocampus 3H-8OH-DPAT bound in the female rats (184+/-16 fmol/mg protein) with respect to the male rats (309+/-41 fmol/mg protein) and to the female sham rats (255+/-20 fmol/mg protein). The forced swim test increases the frontal cortex 5-HT1A receptors in the female rats with respect to the female sham group (40.4+/-5 versus 24.7+/-4 fmol/mg protein, P<0.05). An increased sensibility of the 5-HT1A receptors to depressive-stimulus may be one mechanism underlying the higher prevalence of depression in female. PMID- 14550916 TI - Immunocytochemical study on the distribution of c-myb in the central nervous system of the transgenic mice expressing a human copper/zinc superoxide dismutase mutation. AB - Although our previous study showed the constitutive expression of c-myb in neurons, suggesting that this gene might be involved in the normal function of these cells, there were no reports on the expression pattern of c-myb under pathological conditions. In the present study, we first investigated the changes in c-myb immunoreactivities (IRs) in the central nervous system of the transgenic mice expressing a human copper/zinc superoxide dismutase (Cu/Zn SOD) mutation. The distribution of c-myb was enhanced in the various brain regions of transgenic mice expressing a mutated human Cu/Zn SOD gene. Immunohistochemistry showed intensely stained c-myb IR glial cells with the appearance of astrocytes within the various brain regions of transgenic mice such as the gray matter of the midbrain, medulla oblongata and spinal cord. Even though the exact functions of c myb in the normal and pathological states were not clearly revealed until now, we think that the increase in c-myb expression in the mutant mice could be due to the compensate mechanism of the astrocytes for the reduced defence against superoxide toxicity because the only known function of c-myb was its correlation with the prevention of programmed cell death, which could be deduced from the previous studies. PMID- 14550917 TI - TorsinA protects against oxidative stress in COS-1 and PC12 cells. AB - Dystonia is a highly frequent movement disorder, the pathogenesis of which remains unclear. The cloning of TorsinA, the gene responsible for early-onset dystonia, was a major breakthrough. However, the function of this protein remains unclear. By sequence homology, TorsinA belongs to the ATPases associated with diverse cellular activities-family, many of whose members are chaperones and/or proteases. We report here that in an in vitro model for oxidative stress, H2O2 treatment, overexpression of TorsinA was protective against cell death. COS-1 cells overexpressing TorsinA demonstrated drastically reduced terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling-staining following exposure to H2O2. Furthermore, transfection with TorsinA significantly increased survival of PC12 after H2O2 treatment. To our knowledge, this is the first demonstration that TorsinA protects against oxidative stress. We speculate that a loss of this cellular function in mutant TorsinA may be linked to the pathogenesis of early-onset dystonia. PMID- 14550918 TI - Noradrenaline clearance differs in the frontal cortex and hypothalamus: a dual probe microdialysis study in freely-moving rats. AB - The noradrenergic innervation of the frontal cortex and the hypothalamus arises from different brainstem nuclei and neurones projecting to these two brain regions differ topographically and morphologically. In the present study, we investigated whether there is a difference in noradrenaline clearance in these two brain areas using dual-probe microdialysis in freely-moving rats. Noradrenaline clearance was determined simultaneously in each brain area by measuring the noradrenaline extraction fraction using the 'No Net Flux' method, with both brain regions perfused with the same solutions. The noradrenaline extraction fraction was greater in the frontal cortex than in the hypothalamus (72 and 54%, respectively). This finding suggests that the processes involved in noradrenaline clearance, especially uptake, are more efficient in the former brain area, which could explain the differences in the increase in noradrenaline efflux when these brain areas are challenged with drugs that alter the function of the central nervous system. PMID- 14550919 TI - Pravastatin at 10 mg/day does not decrease plasma levels of either amyloid-beta (Abeta) 40 or Abeta 42 in humans. AB - It has been assumed that statins work as a preventative drug for Alzheimer's disease (AD). Although some epidemiological observations raise doubts to the effectiveness of statins for AD, many in vitro and clinical studies insist on the effectiveness of statins decreasing amyloid-beta (Abeta) levels in medium or blood. To explore the effect of pravastatin on Abeta production, we followed the longitudinal plasma levels of both Abeta 40 and Abeta 42 during the allocation of pravastatin in 46 patients with hyperlipidemia. We found no correlation between plasma cholesterol levels or the decreasing values of total cholesterol and those of Abeta 40 or Abeta 42. Patients having Apolipoprotein E4 (ApoE4) had higher low density lipoprotein levels and lower Abeta 40 levels in plasma, suggesting ApoE4 seems to influence plasma Abeta levels via cholesterol metabolism. PMID- 14550920 TI - Myelin oligodendrocyte glycoprotein is expressed in the peripheral nervous system of rodents and primates. AB - The myelin oligodendrocyte glycoprotein (MOG) is a minor CNS myelin-specific protein that is an important candidate autoantigen in multiple sclerosis. We now report that MOG mRNA transcripts are present in the peripheral nervous system of rodents and primates at levels approximately ten-fold lower than in brain as demonstrated by real time PCR. A major source of this signal are Schwann cells which are also shown to express MOG protein within their cytoplasm in vitro by immunohistochemistry. Expression of MOG by Schwann cells associated with tissue innervation may account for the widespread distribution of low levels of MOG mRNA transcripts, and potentially may provide a source of antigen that can influence the composition and function of the MOG-specific immune repertoire. PMID- 14550921 TI - A case of adult-onset Alexander disease with Arg416Trp human glial fibrillary acidic protein gene mutation. AB - Heterozygous point mutations in the coding region of the human glial fibrillary acidic protein (GFAP) gene have been reported in patients with various forms of Alexander disease (AD). We report a case of genetically confirmed adult-onset AD with palatal myoclonus, pyramidal tract signs, cerebellar signs, and marked atrophy of the medulla oblongata and spinal cord, autonomic dysfunction and heterozygous R416W GFAP mutation. Interestingly, this R416W mutation has also been reported in both infantile and juvenile forms of Alexander disease. The fact that a R416W mutation causes various types of AD suggests that clinical severities of AD are due not only to the different sites and nature of mutations in GFAP, but also to other modifying factor(s). PMID- 14550922 TI - Peroxynitrite induces apoptosis in canine cerebral vascular muscle cells: possible relation to neurodegenerative diseases and strokes. AB - Considerable evidence is accumulating to suggest that in vivo formation of free radicals in the brain, such as peroxynitrite (ONOO-), and programmed cell death (i.e. apoptosis) play important roles in neurodegeneration and stroke. However, it is not known whether ONOO- can induce apoptosis in cerebral vascular smooth muscle cells (CVSMCs). The present study was designed to determine whether or not canine CVSMCs undergo apoptosis following treatment with ONOO-. Direct exposure of canine CVSMCs to ONOO- induced apoptosis in a concentration-dependent manner, as confirmed by means of fluorescence staining, TdT-mediated dUTP nick-end labeling and comet assays. Peroxynitrite treatment resulted in an elevation of [Ca2+]i in the CVSMCs. Peroxynitrite-induced apoptosis may thus be brought about by activation of Ca2+-dependent endonucleases. Although the precise mechanisms by which peroxynitrite induces apoptosis need to be further investigated, the present findings could be used to suggest that ONOO- formation in the brain may play important roles in neurodegenerative processes and strokes via detrimental actions on cerebral microvessels and blood flow. PMID- 14550923 TI - Thiorphan-induced neprilysin inhibition raises amyloid beta levels in rabbit cortex and cerebrospinal fluid. AB - Studies on the pathogenesis of Alzheimer's disease (AD) suggest overproduction of amyloid beta (Abeta) may not be the only pathogenic route to AD. Decreased degradation of Abeta is another possible disease mechanism. Neprilysin is a neutral endopeptidase that has been proposed to be the major enzyme responsible for Abeta degradation. Studies have reported correlations between Abeta deposition and neprilysin activity in the human brain. This study shows that intracerebroventricular infusion of thiorphan, a neprilysin inhibitor, raises cortical and cerebrospinal fluid (CSF) Abeta concentrations in rabbits. Rabbits treated with thiorphan for 5 days had levels of CSF and cortical Abeta40 that were 147 and 142% of the control group, respectively. Results for Abeta42 showed a similar trend. The results indicate that age-related decreases of neprilysin could lead to increased brain concentrations of Abeta, plaque formation, and AD. PMID- 14550924 TI - The role of matrix metalloproteinase-9 in dementia. AB - Neurofibrillary tangles and senile plaques, constituted of extracellular amyloid deposits (Abeta), are the two defining pathological hallmarks of Alzheimer's disease (AD). Inhibiting the synthesis or aggregation of Abeta or increasing its clearance may reduce the detrimental effects of this peptide and consequently improve cognitive functions in patients. Previous studies indicated that metalloproteinases are involved in Abeta degradation and the presence of matrix metalloproteinases (MMP) in AD plaques has been described. In this study, we examined the distribution of a functional polymorphism in the gene for MMP-9, 1562 C-->T, in an independent population of 229 demented and 253 control individuals. We observed a weak protective effect of the high activity allele (T) in apolipoprotein E epsilon4 allele non-bearers (odds ratio=0.5 (95% confidence interval, 0.3-0.9), P=0.04). PMID- 14550926 TI - Immunisation with a synthetic prion protein-derived peptide prolongs survival times of mice orally exposed to the scrapie agent. AB - Several lines of evidence suggest that immunisations may be helpful in the prophylaxis and treatment of neurodegenerative amyloidoses like Alzheimer's disease and prion infections. We used a synthetic prion protein-derived peptide (PrP105-125) and a recombinant PrP fragment (PrP90-230) as antigens for the active immunisation of mice, which were subsequently infected by dietary exposure to the scrapie agent. Immunisation with PrP105-125 prolonged the survival times significantly. In contrast, immunisation with PrP90-230 or adjuvants alone had no effect on the disease development. An epitope mapping of the antibodies raised against PrP90-230 revealed that reactivities against previously defined protective epitopes were either underrepresented or absent. These results point towards the possibility to prevent prion spread via the food chain by vaccinating humans or other species at risk to contract prion diseases. PMID- 14550925 TI - 3-Nitropropionic acid increases frataxin expression in human lymphoblasts and in transgenic rat PC12 cells. AB - Friedreich ataxia (FRDA) is the most common recessive ataxia caused by reduced expression of frataxin, a nuclear encoded mitochondrial protein. In this study we examined the effects of 3-nitropropionic acid (3-NP) on frataxin expression in FRDA patient and control lymphoblasts and in rat pheochromocytoma cell line (PC12) overexpressing human frataxin. Our studies showed an up-regulation of frataxin expression in both FRDA and control lymphoblasts following exposure to 3 NP. In addition, in transgenic frataxin overexpressing cells 3-NP caused an increase of frataxin protein. PMID- 14550927 TI - Effects of nociceptin and analogues of nociceptin upon spontaneous dorsal root activity recorded from an in vitro preparation of rat spinal cord. AB - The 17 amino acid peptide nociceptin has been implicated in pain modulation in the central nervous system. The effects of bath applied nociceptin, and some analogues of nociceptin, upon spontaneous lumbar dorsal root activity have been investigated in an isolated preparation of rat spinal cord. Nociceptin was found to reversibly depress spontaneous dorsal root activity at concentrations of 1.0 microM and 10.0 microM (IC50 2.0 microM), whereas acetyl-nociceptin at concentrations up to 10 microM had no detectable effect. Omission of the last four amino acids (nociceptin 1-13), increased the potency of the effect upon dorsal root activity by approximately 100-fold (IC50 30 nM), but activity was lost when only the first seven amino acids of the nociceptin molecule (nociceptin 1-7) were tested. PMID- 14550928 TI - Comments on "Correlations between brain electrical activities of two spatially separated human subjects", Wackermann et al., Neurosci. Lett. 336 (2003) 60-64. PMID- 14550930 TI - Recognition of childhood depression: personal reminiscences. AB - Prior to 1970, childhood depression was not considered a valid clinical entity by American psychiatrists. One of the early clues was provided in the 1950s by the author's observation of depressive symptoms in children and young adolescents with undescended testicles. This finding was extended to children with several chronic diseases, many of whom exhibited depressive symptoms as well. Eventually, depressive symptomatology was found in children without any physical disorders. This was followed by the introduction of a diagnostic instrument, called the Children's Affective Rating Scale (CARS), later converted into a more formal system called the Child Assessment Schedule (CAS). A provisional classification of childhood depression was published in 1972. Our examination of children with depressive disorders has revealed several modes of family interaction, of which the most important were: separation from important love objects; depreciation and rejection; and affective disorders in parents. Several children with bipolar disorder stimulated our interest in this disorder and led to a pilot study of children of bipolar, lithium-responding parents. Some of these children with bipolar illness had a clear-cut response to lithium and were strong augmenters of the average evoked potentials (EPs). Next, our group investigated the urinary excretion of norepinephrine and its metabolites in chronically depressed children who differed from a normal control group. The foregoing studies, along with major contributions by other child psychiatrists, eventually led to the acceptance of childhood depression as a clinical entity in US psychiatry. The acceptance of juvenile bipolar disorder had to await further research by a new generation of child and adult psychiatrists. PMID- 14550931 TI - Temperament characteristics of child and adolescent bipolar offspring. AB - BACKGROUND: We wished to characterize temperament of children at high risk for bipolar disorder (BD). METHODS: We collected data from the Dimensions of Temperament-Revised (DOTS-R) from 53 biological offspring of at least one parent with BD. RESULTS: Overall, our cohort differed from population means for the DOTS R, having decreased Activity Level-General scores, and increased Approach, and Rhythmicity-Sleep scores. Offspring with psychiatric disorders differed from those without in having decreased Flexibility, Mood, and Task Orientation scores. Temperament profiles for diagnostic categories of BD and attention deficit/hyperactivity disorder were performed in a descriptive manner. LIMITATIONS: Self- or parent-report of temperament was used rather than clinical observation. Temperament characterization was cross-sectional and retrospective rather than prospective and may overlap with clinical diagnoses. CONCLUSIONS: Assessment of temperament may be useful in characterizing bipolar offspring. Decreased flexibility and task orientation, and presence of negative moods may be correlated with development of psychopathology. PMID- 14550932 TI - Toward the development of a Mood Disorders Insight Scale: modification of Birchwood's Psychosis Insight Scale. AB - BACKGROUND: Insight has been defined as: (1) recognition of symptomatology, (2) the ability to attribute symptoms to a mental health disorder, and (3) complying with treatment. Insight is related to medication compliance, course of illness and outcome. Current instruments for measuring insight are limited to those that have been validated primarily in hospitalized patients with psychosis. Our objectives were to develop a reliable and valid self-report scale for use in outpatients or inpatients with mood disorders. Toward this end we made extensive revisions of the Birchwood et al. Insight Scale for Psychosis. METHODS: The scale was developed by modifying items from a previous self-report scale. Specifically, assumptions of hospitalizations, psychosis, and current symptomatology were removed and items related to mood state were added. The scale was included in a battery of measures completed by outpatients and inpatients participating in a study of mood stabilizer medications. RESULTS: Subjects (n=101, 66.3% female, median age 44 years) took approximately 2-3 min each to complete the scale. Overall scores were high (Mean=10.3 out of 12). Reliability was determined using test-retests (r=0.75, n=45). Validity testing was based mainly on clinician ratings (r=0.49, n=69). CONCLUSION: The new scale shows promise as a quick method for assessing insight in patients with mood disorders. PMID- 14550933 TI - Lifetime patterns of depressive symptoms in the community and among primary care attenders: an application of grade of membership analysis. AB - The objective of this study was to describe empirical and natural lifetime patterns of depressive and anxiety symptoms reported by community respondents and primary care attenders. The Grade of Membership model was used to analyze data collected from 716 subjects between 18 and 64 years of age with a lifetime diagnosis of DIS/DSM-III-R Major Depressive Episode. Symptoms of depression, mania, and anxiety (GAD, panic attack, and phobias) were processed. Six prototype categories (pure types) provided the best description of the structure of symptoms included in the analysis. Type I: bipolar depression with marked suicidal behaviour, comorbidity and early onset. Type II: non-melancholic somatisation depression with late onset. Type III: non-melancholic, non-severe bipolar depression with male preponderance. Type IV: depression secondary to anxiety with marked female preponderance. Type V: melancholic depression with suicide ideation. Type VI: melancholic depression with panic attacks and female preponderance. The results support the heterogeneity of the longitudinal symptom pattern of depression and the existence of two time-trend types of comorbid anxiety disorders. PMID- 14550934 TI - Risk for depression: a 6-year follow-up of Finnish adolescents. AB - BACKGROUND: Few longitudinal studies have attempted to identify risk factors in mid-adolescence for subsequent depression in young adulthood. Mid-adolescence is a critical developmental phase for studying vulnerability to depression due to high incidence and prevalence of depression. METHODS: In a longitudinal study, following an urban Finnish community cohort (761 males and 887 females) from age 16, mid-adolescent risk factors for depression at age 22 years were studied. Data were collected by a questionnaire at school at age 16, and by a postal questionnaire at age 22. RESULTS: Of the females 116 (13%) and of the males 69 (9%) had depression (S-BDI) in young adulthood. In multivariate analyses baseline depressive symptoms, low self-esteem, dissatisfaction with academic achievement, problems with the law, poor atmosphere at home and having no close friends predicted subsequent depression. Risk factors for males included more 'externalizing' aspects, for females more 'internalizing' factors. CONCLUSIONS: Mid-adolescence is an important age to study risk for depression, and self reported perceptions of psychosocial well-being have predictive value. Preventive efforts can be selectively targeted at adolescents who have been exposed to identifiable risk factors. PMID- 14550935 TI - Levels of disability in major depression: findings from the Netherlands Mental Health Survey and Incidence Study (NEMESIS). AB - BACKGROUND: Information on the distribution of disability associated with major depression (MD) across different groups of patients is of interest to health policy and planning. We examined the associations of severity and type (a single or recurrent episode) of MD with disability in a Dutch general population sample. METHODS: We used data from the first wave (1996) of the Netherlands Mental Health Survey and Incidence Study (NEMESIS). MD 'severity' and 'type' were diagnosed with the help of the Composite International Diagnostic Interview according to DSM-III-R criteria. SF-36 scores, days ill in bed and days absent from work were taken as indicators of disability. The differences in these variables were studied by means of variance and regression analysis. RESULTS: Recurrent MD was found not to be associated with more disability than single episode MD. Higher 'severity' classes were associated with more disability. However, the degree of disability between 'moderate' and 'severe' MD differed only very slightly. The difference in disability between non-depressed and mildly depressed individuals had a larger effect than between each successive pair of 'severity' classes. CONCLUSIONS: Three groups of MD can be distinguished based on the associated degree of disability: 'mild', 'moderate to severe' and 'severe with psychotic features'. In the future, these groups can be used to describe the distribution of disability in the depressed population. The marked difference between 'mild' MD and no MD suggests that 'mild' cases should be considered relevant. PMID- 14550936 TI - Standardized rater training for the Hamilton Depression Rating Scale (HAMD-17) in psychiatric novices. AB - BACKGROUND: Despite the long and widespread use of the Hamilton Depression Rating Scale (HAMD), standardized reliability studies in inexperienced raters are not available. METHODS: Rater training was carried using three videotaped interviews with depressed patients in 21 psychiatric novices who had negligible previous experience with the HAMD. Chance-corrected coefficients of rating agreement with expert standards (weighted kappa, ICC) were computed for single items and the total score of the HAMD. RESULTS: The results demonstrate sufficiently high interrater reliability (kappa>0.60) for most of the HAMD items and the total score (ICC=0.57-0.73). Three standardized HAMD training sessions seem adequate to establish satisfactory agreement among psychiatric novices. LIMITATIONS: The sample of video-taped interviews and, hence, the generalizability of the results, was restricted. CONCLUSIONS: High inter-rater reliability of the HAMD justifies the use by clinically inexperienced researchers after standardized training. PMID- 14550937 TI - Is there progression from irritability/dyscontrol to major depressive and manic symptoms? A retrospective community survey of parents of bipolar children. AB - BACKGROUND: Although previous studies have discussed age-related changes in the presentation of early onset bipolar illness, the developmental progression of early symptoms remains unclear. The current study sought to trace parents' retrospective report of yearly occurrence of symptoms in a sample of children with and without a diagnosis of bipolar disorder in the community. METHODS: Parents retrospectively rated the occurrence of 37 activated and withdrawn symptoms causing dysfunction for each year of their child's life (mean age 12.6 +/- 6.9). Children were divided into three groups based on parent report of diagnosis by a community clinician: bipolar (n=78); non-bipolar diagnosis (n=38); and well (no psychiatric diagnosis) (n=82). Principal components analysis was performed to understand the relationship among the symptom variables and their potential differences among the three groups as a function of age. RESULTS: Four symptom components were derived and these began to distinguish children with bipolar disorder from the other groups at different ages. Component II (irritability/dyscontrol), which included temper tantrums, poor frustration tolerance, impulsivity, increased aggression, decreased attention span, hyperactivity and irritability, began to distinguish bipolar children from the others the earliest (i.e., from ages 1 to 6). The other components (I, III, and IV) which included symptoms more typical of adult depression (I), mania (III), and psychosis (IV), distinguished the children with a bipolar diagnosis from the others much later (between ages 7 and 12). LIMITATIONS: The data were derived from retrospective reports by parents of their children's symptoms on a yearly symptom check list instrument which has not been previously utilized. Parents' ratings were not validated by an outside rater. Moreover, the children were diagnosed in the community and a formal diagnostic interview was not given. CONCLUSIONS: By parental report, the cluster of symptoms in the irritability/dyscontrol component may characterize the earliest precursors to an illness eventually associated with more classic manic and depressive components that are diagnosed and treated as bipolar disorder in the community. These retrospective survey data suggesting a longitudinal evolution of symptom clusters in childhood bipolar-like illness identify a number of areas for prospective research and validation. PMID- 14550938 TI - The initial prodrome to bipolar affective disorder: prospective case studies. AB - BACKGROUND: The initial prodrome to bipolar disorder has received very little attention to date, with most of the available data only addressing the prodrome to relapse. This study presents several prospective case studies of the initial prodrome to bipolar affective disorder. METHODS: Three patients are presented who developed bipolar disorder during their treatment at the Personal Assessment and Crisis Evaluation Clinic (PACE). They were prospectively interviewed over a 12 month period using standard clinical research interviews. RESULTS: These patients met the criteria for bipolar disorder by the end of the treatment period. Depressive symptoms were the main reason for their first clinical presentation, with mania developing at a later date. Other comorbidities were observed before they were diagnosed with bipolar disorder. LIMITATIONS: The generalisability of our findings was constrained because of the small sample size. Furthermore, our findings are likely to be influenced by the intake criteria used at PACE, a clinic that primarily aims at identifying patients at risk of psychosis rather than bipolar disorder. CONCLUSION: Our study provides information about the initial prodrome to bipolar disorder, which has previously been neglected in research studies. We found there were no prodrome features that clearly distinguished between patients who go on to develop bipolar disorder and those who develop schizophrenia. We hope our prospective data will be the starting point for subsequent studies, with the aim of applying these findings to developing suitable preventative interventions for bipolar disorder. PMID- 14550940 TI - Evolution of the soluble diiron monooxygenases. AB - Based on structural, biochemical, and genetic data, the soluble diiron monooxygenases can be divided into four groups: the soluble methane monooxygenases, the Amo alkene monooxygenase of Rhodococcus corallinus B-276, the phenol hydroxylases, and the four-component alkene/aromatic monooxygenases. The limited phylogenetic distribution of these enzymes among bacteria, together with available genetic evidence, indicates that they have been spread largely through horizontal gene transfer. Phylogenetic analyses reveal that the alpha- and beta oxygenase subunits are paralogous proteins and were derived from an ancient gene duplication of a carboxylate-bridged diiron protein, with subsequent divergence yielding a catalytic alpha-oxygenase subunit and a structural beta-oxygenase subunit. The oxidoreductase and ferredoxin components of these enzymes are likely to have been acquired by horizontal transfer from ancestors common to unrelated diiron and Rieske center oxygenases and other enzymes. The cumulative results of phylogenetic reconstructions suggest that the alkene/aromatic monooxygenases diverged first from the last common ancestor for these enzymes, followed by the phenol hydroxylases, Amo alkene monooxygenase, and methane monooxygenases. PMID- 14550939 TI - Does estrogen enhance the antidepressant effects of fluoxetine? AB - BACKGROUND: While hormone replacement therapy (HRT) has not been shown to be an effective treatment for major depression, preliminary studies suggest that estrogen may potentiate the effect of selective serotonin reuptake inhibitors. METHOD: In an ongoing study, perimenopausal women diagnosed with major depression were randomly assigned to one of three treatment conditions: (1) fluoxetine 10-20 mg alone, (2) estradiol patch 0.1-0.2 mg alone or (3) the combination of fluoxetine 10-20 mg and estradiol patch 0.1-0.2 mg. RESULTS: In the five cases presented here, the combination of fluoxetine and estradiol was most effective, followed by fluoxetine alone and then estradiol alone. LIMITATIONS: These are selected cases from an ongoing study and do not represent statistically significant data. CONCLUSIONS: These preliminary cases suggest that estrogen can enhance the efficacy of antidepressant medication in menopausal women and this adjunctive treatment strategy may be superior to antidepressant or estrogen alone. Further research is needed in elucidating the mechanisms by which estrogen may enhance antidepressant action in perimenopausal women. PMID- 14550941 TI - New concepts of microbial treatment processes for the nitrogen removal in wastewater. AB - Many countries strive to reduce the emissions of nitrogen compounds (ammonia, nitrate, NOx) to the surface waters and the atmosphere. Since mainstream domestic wastewater treatment systems are usually already overloaded with ammonia, a dedicated nitrogen removal from concentrated secondary or industrial wastewaters is often more cost-effective than the disposal of such wastes to domestic wastewater treatment. The cost-effectiveness of separate treatment has increased dramatically in the past few years, since several processes for the biological removal of ammonia from concentrated waste streams have become available. Here, we review those processes that make use of new concepts in microbiology: partial nitrification, nitrifier denitrification and anaerobic ammonia oxidation (the anammox process). These processes target the removal of ammonia from gases, and ammonium-bicarbonate from concentrated wastewaters (i.e. sludge liquor and landfill leachate). The review addresses the microbiology, its consequences for their application, the current status regarding application, and the future developments. PMID- 14550942 TI - Adaptation of Borrelia burgdorferi in the tick and the mammalian host. AB - Borrelia burgdorferi, the causative agent of Lyme disease, shows a great ability to adapt to different environments, including the arthropod vector, and the mammalian host. The success of these microorganisms to survive in nature and complete their enzootic cycle depends on the regulation of genes that are essential to their survival in the different environments. This review describes the current knowledge of gene expression by B. burgdorferi in the tick and the mammalian host. The functions of the differentially regulated gene products as well as the factors that influence their expression are discussed. A thorough understanding of the changes in gene expression and the function of the differentially expressed antigens during the life cycle of the spirochete will allow a better control of this prevalent infection and the design of new, second generation vaccines to prevent infection with the spirochete. PMID- 14550943 TI - Regulation cascade of flagellar expression in Gram-negative bacteria. AB - Flagellar motility helps bacteria to reach the most favourable environments and to successfully compete with other micro-organisms. These complex organelles also play an important role in adhesion to substrates, biofilm formation and virulence process. In addition, because their synthesis and functioning are very expensive for the cell (about 2% of biosynthetic energy expenditure in Escherichia coli) and may induce a strong immune response in the host organism, the expression of flagellar genes is highly regulated by environmental conditions. In the past few years, many data have been published about the regulation of motility in polarly and laterally flagellated bacteria. However, the mechanism of motility control by environmental factors and by some regulatory proteins remains largely unknown. In this respect, recent experimental data suggest that the master regulatory protein encoding genes at the first level of the cascade are the main target for many environmental factors. This mechanism might require DNA topology alterations of their regulatory regions. Finally, despite some differences the polar and lateral flagellar cascades share many functional similarities, including a similar hierarchical organisation of flagellar systems. The remarkable parallelism in the functional organisation of flagellar systems suggests an evolutionary conservation of regulatory mechanisms in Gram-negative bacteria. PMID- 14550944 TI - Bacterial hemoglobins and flavohemoglobins: versatile proteins and their impact on microbiology and biotechnology. AB - In response to oxygen limitation or oxidative and nitrosative stress, bacteria express three kinds of hemoglobin proteins: truncated hemoglobins (tr Hbs), hemoglobins (Hbs) and flavohemoglobins (flavo Hbs). The two latter groups share a high sequence homology and structural similarity in their globin domain. Flavohemoglobin proteins contain an additional reductase domain at their C terminus and their expression is induced in the presence of reactive nitrogen and oxygen species. Flavohemoglobins detoxify NO in an aerobic process, termed nitric oxide dioxygenase reaction, which protects the host from various noxious nitrogen compounds. Only a small number of bacteria express hemoglobin proteins and the best studied of these is from Vitreoscilla sp. Vitreoscilla hemoglobin (VHb) has been expressed in various heterologous hosts under oxygen-limited conditions and has been shown to improve growth and productivity, rendering the protein interesting for biotechnology industry. The close interaction of VHb with the terminal oxidases has been shown and this interplay has been proposed to enhance respiratory activity and energy production by delivering oxygen, the ultimate result being an improvement in growth properties. PMID- 14550945 TI - The rhythm of yeast. AB - Although yeast are unicellular and comparatively simple organisms, they have a sense of time which is not related to reproduction cycles. The glycolytic pathway exhibits oscillatory behaviour, i.e. the metabolite concentrations oscillate around phosphofructokinase. The frequency of these oscillations is about 1 min when using intact cells. Also a yeast cell extract can oscillate, though with a lower frequency. With intact cells the macroscopic oscillations can only be observed when most of the cells oscillate in concert. Transient oscillations can be observed upon simultaneous induction; sustained oscillations require an active synchronisation mechanism. Such an active synchronisation mechanism, which involves acetaldehyde as a signalling compound, operates under certain conditions. How common these oscillations are in the absence of a synchronisation mechanism is an open question. Under aerobic conditions an oscillatory metabolism can also be observed, but with a much lower frequency than the glycolytic oscillations. The frequency is between one and several hours. These oscillations are partly related to the reproductive cycle, i.e. the budding index also oscillates; however, under some conditions they are unrelated to the reproductive cycle, i.e. the budding index is constant. These oscillations also have an active synchronisation mechanism, which involves hydrogen sulfide as a synchronising agent. Oscillations with a frequency of days can be observed with yeast colonies on plates. Here the oscillations have a synchronisation mechanism which uses ammonia as a synchronising agent. PMID- 14550946 TI - Mutational analysis of BARD1 in familial breast cancer patients in Japan. AB - Since BRCA1,which is a breast cancer susceptibility gene, form heterodimers with BARD1, it is speculated that BARD1 mutations might affect the function of BRCA1, contributing to breast carcinogenesis. Thus, in the present study, we have conducted mutational analysis of BARD1 in familial breast cancer patients (n=60) who were tested negative for both BRCA1 and BRCA2 germline mutations. We have been unable to show any clearly deleterious mutations but identified four missence mutations (Ser/Cys 241, Arg/Ser 378, Asn/Ser 470, Val/Met 507), one silent mutation (His/His 506), and one frameshift (in-frame) mutation (1139del21bp). Of these six mutations, one (Asn/Ser 470) was considered as a putative germline mutation since such a mutation was not observed in any of the healthy controls (n=152) but the other five mutations were considered as common genetic polymorphisms since they were frequently observed in the healthy controls. These genetic polymorphisms were further analyzed in their association with breast cancer risk by a case-control study using the population-based breast cancers (n=143) and healthy controls (n=155), which showed that carriers of the variant allele Met at codon 507 are significantly associated with the increased risk of breast cancer (adjusted odds ratio=2.05, 95% confidential interval=1.01 4.16) in postmenopausal women and that the other genetic polymorphisms are not associated with breast cancer risk. These results suggest that BARD1 mutations are responsible for, if any, a very small number of familial breast cancers. Genetic polymorphism of BARD1 (Val/Met 507) could be useful in the selection of postmenopausal women at a high risk for developing breast cancer. PMID- 14550947 TI - DNA adduct formation by the environmental contaminant 3-nitrobenzanthrone in V79 cells expressing human cytochrome P450 enzymes. AB - Diesel exhaust is known to induce tumours in animals. Of the compounds found in diesel exhaust 3-nitrobenzanthrone (3-NBA) is particularly a powerful mutagen. Recently we showed that 3-NBA is genotoxic in vivo in rats by forming specific DNA adducts derived from nitroreduction. In this study a panel of genetically engineered V79 Chinese hamster cell lines expressing various human cytochrome P450 (CYP) enzymes (CYP1A1, CYP3A4) and/or human NADPH:CYP oxidoreductase (CYPOR) was used to identify CYP enzymes involved in the metabolic activation of 3-NBA. We analyzed the formation of specific DNA adducts by 32P-postlabelling after exposing cells to 1 microM 3-NBA. A similar pattern with a total of four distinct 3-NBA-DNA adducts was found in all cells, identical to those detected previously in DNA from rats treated with 3-NBA in vivo. Total adduct levels ranged from 75 to 132 using nuclease P1 and from 103 to 220 adducts per 10(8) nucleotides, using butanol enrichment. Comparison of DNA binding between different V79MZ derived cells revealed that human CYPOR and CYP3A4 were involved in the metabolic activation of 3-NBA. Furthermore, dose-dependent high adduct levels were detected after exposure to 0.01, 0.1 or 1 microM 3-NBA in the subclone V79NH which exhibits high activities of nitroreductase and N,O-acetyltransferase. Our results suggest that nitroreduction is the major pathway in the human bioactivation of 3 NBA. Moreover, acetylation of the initially formed N-hydroxy arylamine intermediates may contribute to the high genotoxic potential of 3-NBA. PMID- 14550948 TI - Evaluation of colloid size for sentinel nodes detection using radioisotope in early gastric cancer. AB - The purpose of this study was to investigate the relationship between colloid size and the detection of sentinel nodes (SN) in early gastric cancer. Three size of 99mTechnetium-tin colloids (500, 100 and 50 nm) were preoperatively injected into the submucosa under endoscopic control. Lymph node metastasis and micrometastasis was examined. RI-uptake in the hottest nodes and the total RI uptake in the hot nodes were highest in the size of 100 nm. At least one lymph node metastasis, including micrometastasis, was included in the hot nodes. RI labeled colloid size was one of the important factors to detect SN in early gastric cancer. PMID- 14550949 TI - Down's syndrome-associated Single Minded 2 gene as a pancreatic cancer drug therapy target. AB - We report here a pancreatic cancer drug therapy utility of a gene involved in Down's syndrome. Single Minded 2 gene (SIM2) from Down's Syndrome Critical Region was expressed in pancreatic cancer-derived cell lines as well as in tumor tissues, but not in the normal pancreas. A related member of the SIM family, SIM1, did not show similar specificity. Inhibition by antisense technology of one of the isoforms of SIM2, the short-form (SIM2-s) expression in the CAPAN-1 pancreatic cancer cell line, caused a pronounced growth inhibition and induced cell death through apoptosis. The specificity of antisense was inferred from inhibition of SIM2-s mRNA but not the related members of SIM family. In view of the high mortality rate of pancreatic cancer patients, these findings have important implications for the future of pancreatic cancer treatment. PMID- 14550950 TI - The comparison of the prognosis between Epstein-Barr virus (EBV)-positive gastric carcinomas and EBV-negative ones. AB - The relationship between the degree of lymphocytic infiltration into the tumor and the prognosis has not been completely evaluated between Epstein-Barr virus (EBV)-positive and -negative gastric carcinoma (GC). Although the average numbers and the grades of the infiltrating CD8+T cells, natural killer cells, dendritic cells, Ki67-positive cells were significantly greater in EBV-positive GCs than in -negative GCs, there was no significant survival improvement in EBV-positive group. These findings suggest that the infiltration of lymphocytes in the EBV positive GC does not necessarily meant better prognosis and that the EBV status is not a significant prognostic factor in the patients with gastric cancer. PMID- 14550951 TI - Biphasic modulation of cell proliferation by quercetin at concentrations physiologically relevant in humans. AB - Optimal in vitro conditions regarding quercetin solubility and stability were defined. Using these conditions, the effect of quercetin on proliferation of the colon carcinoma cell lines HCT-116 and HT29 and the mammary adenocarcinoma cell line MCF-7 was investigated. For the colon carcinoma cell lines, at relatively high concentrations, a significant decrease in cell proliferation was observed, providing a basis for claims on the anti-carcinogenic activity of quercetin. However, at lower concentrations, a subtle but significant stimulation of cell proliferation was observed for all cell lines tested. These results point at a dualistic influence of quercetin on cell proliferation that may affect present views on its supposed beneficial anti-proliferative effect. PMID- 14550952 TI - A relationship between Matrix metalloproteinase-1 (MMP-1) promoter polymorphism and cervical cancer progression. AB - Single nucleotide polymorphism (SNP) of the promoter region of MMP-1 (at -1607 bp) creates Ets binding sites, and correlations between this SNP and cancer susceptibility have been reported for various cancers. In this study, we genotyped the SNP in 23 cervical intraepithelial neoplasias (CIN) and 86 cervical cancer specimens. We found a correlation between promoter polymorphism and MMP-1 expression, and that this SNP was correlated with the clinical stage of cervical cancer. These findings suggested that SNP of MMP-1 promoter might influence the ability in cervical cancer invasion via transcriptional activity of this gene. PMID- 14550953 TI - Extracellular matrix protein 1 (ECM1) is over-expressed in malignant epithelial tumors. AB - The extracellular matrix protein 1 (ECM1) is a secreted protein that has been implicated with cell proliferation, angiogenesis and differentiation. In the present study, we used immunohistochemical staining to examine the expression of ECM1 in a panel of human tumors and found that it was closely correlated with some types of tumors including: invasive breast ductal carcinoma (83%), esophageal squamous carcinoma (73%), gastric cancer (88%) and colorectal cancer (78%). Significantly, ECM1expression was correlated with the metastatic properties of the tumors. Primary breast cancers that had formed metastases were 76% positive while those that had not metastasized were only 33% positive. ECM1 expression was also correlated with PCNA a marker for proliferation, but not with CD34, a marker for endothelial cells. These results indicate that ECM1 tends to be preferentially expressed by metastatic epithelial tumors. PMID- 14550954 TI - Overexpression of Reg IV in colorectal adenoma. AB - Identification of molecular markers associated with colorectal adenoma may uncover critical events involved in the initiation and progression of colorectal cancer. Our previous studies, mainly based on suppression subtractive hybridization, have identified Reg IV as a strong candidate for a gene that is highly expressed in colorectal adenoma when compared to normal mucosa. In this study, we sought to determine the mRNA expression of Reg IV in colorectal adenoma, in comparison with normal colorectal mucosa and carcinoma in multiple samples. Semi-quantitative RT-PCR was performed in 12 colorectal adenomas and 10 concurrent carcinomas. Reg IV mRNA level was higher in all adenomas (12/12) (p=0.001) and in 9/10 concurrent colorectal carcinoma (p=0.021) when compared to paired normal colorectal mucosa. Northern blot analysis further confirmed these results. In situ hybridization with digoxigenin (DIG)-labeled cRNA was performed in 32 colorectal adenomas with varying degree of dysplasia. Compared with paired normal tissues, Reg IV was overexpressed in 74% (14/19) adenomas with mild or moderate dysplasia and 100% (13/13) cases of adenoma with severe dysplasia. In addition, higher levels of Reg IV mRNA was consistently scored in regions with more severe dysplasia within the same adenoma sample displaying varying degree of dysplasia. The strongest staining was seen within carcinomoutous areas of the 12 adenoma cases (p=0.002). Our results support that overexpression of Reg IV may be an early event in colorectal carcinogenesis. Detection of Reg IV overexpression may be useful in the early diagnosis of carcinomatous transformation of adenoma. PMID- 14550955 TI - Disseminated cancer cells in the blood and expression of sialylated antigen in gastric cancer. AB - BACKGROUND: In gastric cancer, disseminated cancer cells (DCC) can be detected in peripheral blood using bio-molecular techniques. It is known that patients having DCC exhibit a high occurrence of postoperative relapse in gastrointestinal cancer. However, more than half of gastric cancer patients having positive DCC do not show cancer relapse. Sialylated Lewis antigens are considered to be crucial molecules in the metastasis of disseminated cancer. The current study investigated whether combination analysis of DCC and sialylated Lewis antigen are useful in estimating the recurrence risk of gastric cancer. PATIENTS AND METHODS: Subjects were 106 consecutive gastric cancer patients who underwent curative gastrectomy. DCC in the peripheral blood were detected using the carcinoembryonic antigen (CEA)-mRNA by RT-PCR method. Sialylated Lewis antigen expression (sLeA and sLeX) of the primary tumor was assessed immunohistochemically. RESULTS: Of 106 gastric cancer patients, 43 (40%) were positive for DCC. Immunohistochemically, 53 (50%) and 49 (46%) patients were positive for sLeA and sLeX, respectively. The presence of DCC did not correlate with sLeA and sLeX expression in gastric cancer. Postoperative tumors were present in 19 patients (7 hematogenous and 12 non-hematogenous), 12 of which were positive for DCC. Six sLeA-positive patients (26%) with DCC and 13 sLeX-positive patients (57%) with DCC suffered from postoperative recurrence of gastric cancer. The p value of CEA mRNA and sLeX combination analysis was more significant (p<0.01) than that of CEA mRNA alone (p=0.02). CONCLUSION: Analyzing both DCC and sLeX expression in gastric cancer may enable more accurate prediction of postoperative recurrence. PMID- 14550956 TI - Low expression of sodium iodide symporter identifies aggressive thyroid tumors. AB - A decreased radioiodine uptake is frequently detected in differentiated thyroid carcinomas (DTC) and is associated with high recurrence rate and reduced survival. We investigated the correlation between NIS mRNA expression levels in the primary tumor and patient outcome using a quantitative real-time RT-PCR method. NIS expression was decreased in 17 DTC (21.04+/-39.66 pg Eq) compared to four autoimmune thyroid disease (180.51+/-92.63 pg Eq) and 14 normal tissues (75.71+/-66.98 pg Eq) (p<0.0001). The 17 thyroid differentiated carcinoma patients were submitted to surgery complemented by radioiodine ablation and had at least 24 months of follow-up, under levothyroxine continued suppressive therapy. According to their outcome, we could characterize a group of papillary carcinoma patients with aggressive carcinomas, whose NIS mRNA levels were markedly lower than a group with non-aggressive carcinomas (0.62+/-0.79 versus 54.87+/-53.79; p<0.005). We suggest that the quantification of NIS mRNA relative levels in the primary tumor may predict poor outcome. PMID- 14550957 TI - Microbial dynamics of purulent nasopharyngitis in children. AB - This review presents the microbiological dynamic and therapeutic options in the management of purulent nasopharyngitis (NPT). The nasopharynx (NP) of healthy children is generally colonized by relatively non-pathogenic aerobic and anaerobic organisms, some of, which possess the ability to interfere with the growth of potential pathogens. Conversely, carriage of potential respiratory aerobic pathogen such as Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis, as well as some anaerobic bacteria (Peptostreptococcus, Fusobacterium and Prevotella spp.) increases during purulent NPT. The development of purulent NPT in children is associated with the pre-existing colonization by potential pathogens and the absence of interfering organisms in the NP. Controversy exists regarding the management of NPT as no conclusive evidence exists to date that the administration of antimicrobials will shorten the illness. PMID- 14550958 TI - Obstructive sleep apnea syndrome due to adenotonsillar hypertrophy in infants. AB - OBJECTIVE: Adenotonsillar hypertrophy (ATH) is the leading cause for obstructive sleep apnea syndrome (OSAS) in children. The peak age for adenoid and tonsillar hypertrophy and related OSAS is 3-6 years. It has been suggested that OSAS due to ATH is extremely rare in infants. The purpose of the present study was to delineate OSAS due to ATH in infants. METHODS: Twenty-nine consecutive infants <18 months of age who underwent polysomnography (PSG) and were diagnosed with OSAS due to ATH were studied. A pediatric sleep questionnaire was completed by parents of all infants. Information regarding demographic variables, referring physician specialty, assessment of infant's development and recurrence of symptoms post treatment was collected. Pre and post surgery body weight and developmental assessment by parents were evaluated. RESULTS: The mean age of adenotonsillectomy was 12.3+/-3.9 months with the mean duration of symptoms ranging 6.2+/-3.0 months. 24% of the infants were born preterm. Snoring was the most common finding and appeared in all infants. Other symptoms were sleep apnea (72%), frequent movements during sleep (69%), mouth breathing (62%) and recurrent awakenings (38%). During the preoperative period, mean body weight decreased from the 67th+/-25 percentile to the 42nd+/-32 percentile (P<0.00001). 14/29 (48%) of the infants dropped two or more major percentiles before treatment. A significant weight gain to the 59th+/-31 percentile was observed post surgery (P<0.0001). 5/29 (17%) infants were considered by their parents as having a developmental delay preoperatively, which resolved in 3/5 (60%) post treatment. Clinical symptoms resolved or improved significantly after surgery. Recurrence of symptoms was documented in 6/23 (26%) of infants. CONCLUSIONS: Infantile OSAS due to hypertrophic adenoids and tonsils does occur in infants. Unique characteristics for this age group include: male predominance, high incidence of preterm infants, failure to gain weight and high recurrence rate after surgery. Otolaryngologists and pediatricians should be aware to the existence of the "early OSAS" in small infants. PMID- 14550959 TI - Monosyllable speech perception of Japanese hearing aid users with prelingual hearing loss: implications for surgical indication of cochlear implant. AB - OBJECTIVE: The monosyllable speech perception ability after years of educational intervention was compared between prelingually deafened pediatric hearing aid users and their cochlear implant counterparts. DESIGN: An open-set monosyllabic speech perception test was conducted on all subjects. The test required subjects to indicate a corresponding Japanese character to that spoken by the examiner. Fifty-two subjects with prelingual hearing impairment (47 hearing aid users and 5 cochlear implant users) were examined. RESULTS: Hearing aid users with average pure-tone thresholds less than 90 dB HL demonstrated generally better monosyllable perception than 70%, which was equivalent or better performance than that of the cochlear implant group. Widely dispersed speech perception was observed within the 90-99 dB HL hearing-aid user group with most subjects demonstrating less than 50% speech perception. In the cluster of >100 dB HL, few cases demonstrated more than 50% in speech perception. The perception ability of the vowel part of each mora within the cochlear implant group was 100% and corresponding to that of hearing aid users with moderate and severe hearing loss. CONCLUSION: Hearing ability among cochlear implant users can be comparable with that of hearing aid users with average unaided pure-tone thresholds of 90 dB HL, after monosyllabic speech perception testing was performed. PMID- 14550960 TI - Transnasal endoscopic dacryocystorhinostomy for the treatment of lacrimal pathway stenoses in pediatric patients. AB - Lacrimal pathway stenoses in pediatric patients are uncommon after the age of 1 year. Various conservative treatments (massages, hot compresses, topical antibiotics, probing and intubation of the lacrimal pathways) are generally used to resolve these disorders. When non-invasive procedures fail, dacryocystorhinostomy (DCR) is indicated. While this surgical procedure has traditionally been performed by an external approach, a less-invasive endonasal technique has recently been advocated. We herein review a series of six children affected by lacrimal pathway stenosis who were treated at the Department of Pediatric Otorhinolaryngology, Spedali Civili, Brescia (Italy) between November 2000 and January 2002. All patients underwent successful transnasal endoscopic DCR and, to date, no re-stenoses have occurred. A review of the literature focusing on diagnostic work-up and different therapeutic options is also presented. PMID- 14550961 TI - Noticeable differences in bacterial defence on tonsillar surfaces between bacteria-induced and virus-induced acute tonsillitis. AB - OBJECTIVE: Oral and pharyngeal cavities harbor a commensal bacterial flora which is kept in check by several innate and acquired agents. In this study, we focused on the proportions in which some antibacterial moderators (lysozyme, lactoferrin, IgG and S-IgA) coat the tonsillar surface bacteria in healthy individuals, in patients with acute tonsillitis (AT) culture-positive for Streptococcus pyogenes, and in patients with infectious mononucleosis (IM) caused by Epstein-Barr virus (EBV). METHODS: Bacterial samples were collected for aerobic culturing and immunocytochemical evaluation from the tonsillar surfaces of eight healthy individuals (four males, four females; age range 16-22 years), eight patients with current AT (two males, six females; age range 16-29 years) and seven patients with IM (four males, three females; age range 15-21 years). The immunocytochemical assay was based on gold-labeled antiserum to human lysozyme, lactoferrin, IgG and S-IgA followed by gold particle tracing in the transmission electron microscope. RESULTS: During AT, a significant increase in lysozyme coating (P<0.05) and lactoferrin coating (P<0.0005) of the bacteria was noted, whereas the S-IgA coating was significantly reduced (P<0.0005). During IM infection, a significant increase in lactoferrin coating was noted (P<0.0005) whereas immunoglobulin coating was significantly reduced (IgG P<0.025; S-IgA P<0.0005) compared with healthy controls. During IM, all antibacterial moderators evaluated were significantly reduced compared with the situation during AT. CONCLUSIONS: Noticeable changes in the local innate and acquired bacterial defence system were observed during tonsillar infections, particularly during IM. PMID- 14550962 TI - Binaural interaction of bone-conducted auditory brainstem responses in children with congenital atresia of the external auditory canal. AB - Bilateral bone-conducted auditory brainstem responses (BC-ABRs) were recorded in children with atresia of the external auditory canal bilaterally (AECB) in order to compare the response characteristics to normal hearing adults. The binaural interaction component (BIC) of the ABR occurs when the sum of the monaural-evoked ABR amplitudes are different in amplitude when compared to the binaural-evoked ABR amplitude. Previous electrophysiological work from our lab has shown that children with AECB lateralize bone-conducted (BC) sound. Furthermore, we have found in normal-hearing adults that BICs exist using BC clicks. In adults, BC-BIC occurred in the latency region corresponding to waves IV-VI, whereas for children with AECB corresponding peak latencies occurred earlier. Same as normal-hearing adults, BC-ABR IV-V complex peak amplitudes for sum of the BC-monaural right and BC-monaural left ears were different from binaural response amplitude. Individual peak latencies were similar in children with AECB when compared to normal-hearing adults except for shorter latencies for BIC. These results indicate that: (1) BC BI is present in children with AECB as well as normal-hearing adults; (2) the gross response properties of BIC are similar in children with AECB and normal hearing adults; (3) fitting of a bilateral BC hearing aid might be a feasible method to optimize binaural hearing and sound lateralization. PMID- 14550963 TI - Accuracy and feasibility of daily infrared tympanic membrane temperature measurements in the identification of fever in children. AB - BACKGROUND: Fever is an important physical sign in infectious diseases in childhood. Daily assessment of fever may be used to monitor the occurrence of infections for research purposes. The infrared tympanic membrane thermometer (ITT) seems ideal for measuring central body temperature. Its accuracy, however, is still debated, and the feasibility of daily temperature measurements with an ITT has not been described. OBJECTIVE: To determine the accuracy and feasibility of daily infrared tympanic membrane temperature measurements at home. METHODS: Tympanic membrane temperature was measured by the Braun Pro 3000, rectal temperature by the Omron MC 63. To establish the accuracy of the ITT, rectal and tympanic membrane temperature measurements were performed in 41 children, aged 2 10 years. To establish the feasibility of daily infrared tympanic temperature measurements at home, parents of 21 children performed daily measurements for 2 weeks. RESULTS: With fever defined as a body temperature of > or =38.0 degrees C, sensitivity was 93.3%, specificity 92%, positive predictive value 87.5%, negative predictive value 95.8%, and the concordance rate 92.5%. Most of the parents found the instructions for the use of the ITT clear (93%) and the ITT easy to use (86%). During the test period of 2 weeks the technique of the tympanic membrane temperature measurements remained adequate in 93% of the parents. CONCLUSION: The tympanic membrane temperature measured by the Braun Pro 3000 accurately reflects rectal temperature, validly assesses the presence of fever in children, and is easy to use. The Braun ITT therefore is an adequate tool to assess fever and may be used both in a clinical setting and for research purposes. PMID- 14550964 TI - A model of two-stage newborn hearing screening with automated auditory brainstem response. AB - Our purpose was to evaluate a two-stage newborn hearing screening program using automated auditory brainstem response (AABR) before discharge and to describe our follow-up program. This study used 4085 infants born in the Seirei-Hamamatsu and Mikatahara General Hospitals during a 2-year period. The initial screening test was performed 2 or 3 days after birth at an intensity of 35 dBnHL. For the infants who were referred from this test, the re-screening test was performed 5 or 6 days after birth. Diagnostic work-up with auditory brainstem response (ABR), otoacoustic emissions (OAE), and a conditioned orientation reflex audiometry (COR) test were performed by the age of 3-6 months. The referral rate was 1.20% (49/4085 infants) in the first test and 0.71% (29/4085 infants) in the two-stage screening. The two-stage screening procedure was able to reduce the false positive rate from 0.83 to 0.34%. The incidence of bilateral and unilateral congenital hearing loss diagnosed by ABR was 8/4085 (0.20%) infants and 7/4085 (0.17%) infants, respectively. One infant with congenital cytomegalovirus infection, who passed the two-stage AABR tests, was diagnosed with hearing loss 1 month after birth, using ABR. The two-stage measurement of AABR is effective and time efficient due to significant decreases in the referral rate and the false positive rate. PMID- 14550965 TI - Postnatal change on the location of Ostmann's fatty tissue in the region lateral to Eustachian tube. AB - OBJECTIVE: To delineate a possible role of protective function by Ostmann's fatty tissue (OF) in the lateral region to Eustachian tube (ET). STUDY DESIGN: Morphometric and histopathological examination on human temporal bones. METHODS: OF in the region lateral to the cartilaginous portion of the ET between the ET lumen and the tensor veli palatini muscle (TVP) was studied histopathologically and morphometrically by analyzing a series of vertically cut histologic sections for the ET structures. Sections were obtained from 25 individuals with age ranging from 33 gestational weeks to 38 years at the time of their death. RESULTS: In fetus and neonates, the region lateral to ET was filled with mesenchyme. OF appeared first in this region at 2-3 weeks postnatally, and increased in volume with development of the ET. In the cases of older children and adults (Group-B), the ratio of the volume of OF in the region to that of ET lumen (OF/L) was significantly higher in the posterior half than in the anterior half of the cartilaginous portion of ET. However, in the cases of infants and younger children (Group-A), there was no significant difference in the ratio between the two regions. In the posterior half of the cartilaginous portion of ET, OF/L was significantly higher in Group-B than in Group-A, while in the anterior half, there was no significant difference in the ratio between two age groups (Group-A and B). CONCLUSIONS: In Group-B, the dominant OF in the area of the posterior half of the cartilaginous region, which includes the narrowest portion of ET, may be responsible for restoring the ET lumen back to its closed static condition after active tubal opening by contraction of TVP. This is thought to be important for protection of the middle ear. On the other hand, in Group-A, poor OF near the narrowest portion of ET may cause insufficient restoration of the ET lumen and increased risk of developing otitis media (OM). PMID- 14550966 TI - Intraparotid facial nerve solitary plexiform neurofibroma: a first paediatric case report. AB - Solitary plexiform neurofibromas of the intraparotid facial nerve are extremely rare tumours that can present a diagnostic challenge. Knowledge of their existence is key to its proper management. Only two cases have been reported in the English literature. We describe the first paediatric case of an intraparotid facial nerve solitary plexiform neurofibroma. PMID- 14550967 TI - Lower airway papillomatosis in children. AB - Laryngeal papilloma in children is a frequent disease caused by human papilloma virus (HPV) type 6 or type 11. This disease has a tendency to recur and the changes are histologically benign. In some cases papilloma may affect the lower levels of the respiratory tract. In this study, among 90 patients treated for laryngeal papillomatosis, in four children papilloma of trachea, bronchi and lung tissue were detected in endoscopic and radiological examination. This constitutes 4.4% of all patients. Compact nodules and acquired cysts between 5 and 50 mm long were found in chest X-rays and in computerised tomography. These cysts appeared from 4 to 8 years after establishing a diagnosis of laryngeal papilloma, and 1 year after recognising papilloma in the trachea. In all four children the presence of nodules and cysts in the lungs was preceded by recurrent pneumonia, emphysema or atelectasis of the lungs. All children with laryngeal papillomatosis should have a chest X-ray. Detection of acquired cyst-like changes in lung tissue in children with laryngeal papillomatosis is a warning of future papilloma in the trachea and bronchi, with involvement of lung tissue. In differential diagnosis of these changes in the lungs we should take into consideration the presence of papilloma in the bronchi. A prognosis of papillomatosis in the lower airways in children is always serious. PMID- 14550968 TI - Migrating foreign body in the bronchus. AB - Tracheobronchial foreign bodies especially in infants and children is fraught with respiratory obstruction and even can lead to death if not intervened in time. The patients can present in variable forms but clinical suspicion and management for such cases is an art itself. We report a case of an unusual foreign body in a child of 1 year showing migratory movements from one main bronchus to other. PMID- 14550969 TI - Deletion mapping of split hand/split foot malformation with hearing impairment: a case report. AB - Split hand/split foot malformation (SHFM), which typically appears as lobster like limb malformation, is a rare clinical condition caused by a partial deletion of chromosome 7q. Hearing impairment sometimes accompanies syndromic SHFM cases; a case of inner and middle ear malformation with SHFM is described in this report. We conducted a genetic evaluation of this patient and found a deleted region that overlaps a previously reported locus of SHFM as well as a DFNB14 locus that can cause nonsyndromic hearing impairment by autosomal recessive inheritance. PMID- 14550970 TI - Auditory neuropathy in siblings with Waardenburg's syndrome. AB - We report two siblings with a family history of Waardenburg's syndrome (WS) for whom the audiological profile corresponds to auditory neuropathy (AN). They have; (1) bilateral severe to profound hearing loss, (2) robust oto-acoustic emissions (OAEs) in both ears, and (3) no auditory evoked responses at 95 dBnHL bilaterally. Electrocochleography (ECochG) and auditory middle and late latency potentials were performed in one of the children. Results showed cochlear and neural activities in both ears. Central auditory responses were not conclusive. These children did not have any history of neonatal illness and one child was diagnosed with AN at the age of 3 weeks and the other at the age of 11 months. PMID- 14550971 TI - The tongue-base suspension using Repose bone screw system in a child with Simpson Golabi-Behmel syndrome. Case report. AB - Obstructive sleep apnea syndrome is one of the most frequent manifestations of upper airway obstruction. Several approaches have been proposed to solve this problem with the aim to avoid tracheotomy especially in children. We observed a child affected by Simpson-Golabi-Behmel syndrome presenting severe OSAS. He underwent tongue-base suspension using the Repose system with significant improvement of the polysomnographic parameters and of the clinical conditions. Tongue suspension with this technique effectively prevent tongue-base collapse, avoiding tracheotomy or increased morbidity caused by a more invasive procedure. Besides it does not interfere with the patient's cranio-facial growth and future occlusion. PMID- 14550972 TI - [Novel applications for hematopoietic growth factors?]. PMID- 14550973 TI - [Acute gastro-enteritis in children in France: estimates of disease burden through national hospital discharge data]. AB - OBJECTIVES: To estimate the burden of hospitalized infectious gastroenteritis of children younger than 5 years of age and associated costs. METHODS: We analyzed 1997 hospital discharges with a primary diagnosis of gastroenteritis or a secondary diagnosis of gastroenteritis with gastroenteritis symptoms or complications as primary diagnosis and compared the deaths with those of the national mortality data. RESULTS: Gastroenteritis was associated with 51,125 hospitalizations which accounted for 11.4% of hospitalization discharges for this age group and an annual rate of 1,385 per 100,000 children <5-year-old. Most gastroenteritis (56%) were registered as "probably infectious", 36% as "viral" (43% of which were coded "rotavirus") and 8% as "bacterial" (of which 60% were coded "Salmonella"). The seasonal peak was winter for rotaviral, viral and "probably infectious" gastroenteritis, summer for those related to salmonellosis. Incidence increased inversely with age: 3606/100,000 infants <1-year-old, 257/100,000 4-year-old children. Complications (especially dehydration) were observed in 21% of viral gastroenteritis and 17% of bacterial gastroenteritis. At least, 14 deaths were found in both hospital discharge and mortality data. The mean duration of stay (3.2 days) was significantly higher in infants <1-year-old, viral etiology, association with complications or bronchiolitis. The costs of hospitalization could be estimated to 62 million Euros. DISCUSSION: Our results are similar to those obtained in other developed countries. Despite variations in encoding the discharge reports, data has proven to be effective to describe national trends for this health event. Our study indicates that the public health burden and economic impact of prevention and control measures can be monitored through hospital discharge surveillance. PMID- 14550974 TI - [Access to early speech therapy in Gironde: evaluation of a language-screening campaign for 4-year-old children (1999-2001)]. AB - OBJECTIVE: The objective of this article was to evaluate the results of a prevention campaign on the access to early speech therapy for 4-year-old children. POPULATION AND METHODS: A prevention campaign of speech disorders was conducted in Gironde area in 1999, offering a systematic screening by ERTL4 test and free speech therapy. The campaign results were evaluated by quality and efficiency indicators. RESULTS: In 1999, 14037 children born in 1995 were included and 1363 children randomly selected from the schools benefited from the campaign. One child out of four failed in the screening test by ERTL4. The percentage of the assessment and speech therapy in the children of the campaign (16.0 and 8.2%) were significantly higher than the other children of the area (8.9 and 6.7%). CONCLUSION: The campaign evaluation has demonstrated its capacity to access to speech therapy for children language disorders, allowing gains of frequency and earlier treatment. PMID- 14550975 TI - [Study of correlations between antibiotics prescriptions and guidelines in a neonatal intensive care unit]. AB - The increasing use of wide spectrum antibiotics has been reported to be associated with a greater prevalence of multi-resistant bacteria. OBJECTIVES: The aims of this study were to survey the use of antibiotics and to evaluate the correlations between patterns of prescription of antibiotics and prescription guidelines in a neonatal intensive care unit. MATERIAL AND METHODS: In this 6 month study, all newborns admitted to the NICU and treated with antibiotics were included. Data regarding criteria of antibiotic prescription, length of treatment, and criteria of withdrawing treatment were collected. The correlation between prescriptions and guidelines was evaluated a posteriori by a non prescriber physician. One hundred and sixteen newborns were included, of whom nine had received antibiotics on more than one occasion. Mean gestational age was 33.5 weeks. In 82% of cases, the reason for hospitalisation was respiratory distress syndrome. RESULTS: Patients received systemic antibiotics for primary infection (78%), nosocomial infection (17%) and postsurgical prophylaxis (5%). Suspected foeto-maternal infections (SFMI) were the dominant features of primary infection (96%). In 49% of cases, suspected infection was not proven and justified withdrawal of treatment within 3 days. Sixty percent of nosocomial infections occurred in newborns with gestational ages of less than 28 weeks. Bacterial criteria were decision-making factors only in nosocomial infections. An absence of observance of guidelines occurred in 9% of treated newborns, and in most cases involved excessive length of treatment. CONCLUSION: These results show: (1) the majority of antibiotic prescriptions were for not proven SMFI; (2) a low rate of nosocomial infections; (3) the predominance of nosocomial infections in premature newborns; (4) less than 10% of non-observance of guidelines. It appears necessary to develop more precise guidelines to limit antibiotic use and to evaluate them regularly. PMID- 14550976 TI - [Nosocomial Burkholderia cepacia outbreak in an intensive pediatric care unit]. AB - BACKGROUND: We report an outbreak of Burkholderia cepacia respiratory tract infection and colonization in an intensive pediatric care unit.P PATIENTS AND METHODS: Between February and December 1999, B. cepacia was isolated from five children hospitalized in this unit. We reviewed the charts of the patients, evaluated the antiseptics use and the disinfection practices for reusable patient care equipment. An environmental study was conducted and comparison of B. cepacia was performed with genotypic method (RAPD). RESULTS: All patients were mechanically ventilated and had received large spectrum antibiotics. The disinfection procedure for reusable equipment was not respected and some single dose of antiseptics solutions were used for several patients. B. cepacia was not found in 34 environmental samples. The RAPD assay revealed that all five isolates had identical DNA profiles. CONCLUSION: Despite the investigation the source of the B. cepacia clone in this nosocomial outbreak remained unknown, but antiseptics use and disinfection practices were revised. No new B. cepacia infections were identified after control measures were implemented. PMID- 14550977 TI - [HPA-5b neonatal allo-immune thrombocytopenia: two cases described in Tunisia]. AB - Alloimmune thrombocytopenia is due to feto-maternal incompatibility in the HPA systems and is usually considered in the diagnosis of neonatal thrombocytopenia after other causes have been excluded. We report on two Tunisian observations of alloimmune neonatal thrombocytopenia due to anti-HPA-5b (Bra) antibodies. CASE REPORT: Two neonates presented at birth with a thrombocytopenic purpura unexplained by usual causes of neonatal thrombocytopenia. Alloimmune neonatal thrombocytopenia was diagnosed by the determination of parental and neonatal platelets antigens phenotypes and by the presence of HPA-5b (antiBra) antibodies in maternal sera. A favourable evolution was obtained after maternal platelet transfusions. CONCLUSION: Alloimmune neonatal thrombocytopenia is a serious affection, which exposes to intracranial haemorrhage. These observations of HPA-5 neonatal alloimmunisation in Tunisia provide additional information on the geographic distribution of the disease and its prognosis. PMID- 14550978 TI - [Juvenile myelomonocytic leukaemia, xanthoma, and neurofibromatosis type 1]. AB - The triple association of leukemia, xanthogranulomas, and type 1 neurofibromatosis was first described in 1958. Most leukemias were juvenile myelomonocytic leukemias (JMML), usually called juvenile chronic myelogenous leukemia. We describe a 22-month-old female child with neurofibromatosis 1, xanthomagranulomas, and a JMML. Her mother and her brother also had cutaneous cafe-au-lait spots. Our patient was treated with mercaptopurine and improved. However, 9 months later she experienced a blastic transformation. The presence of xanthomagranulomas and NF1 in a young child should alert to a possible development of JMML, especially in patients with a family history of NF1. PMID- 14550979 TI - [Peritoneal hydatidosis in children. Report of a historical case]. AB - Peritoneal hydatidosis is a rare localisation of hydatic disease, most often secondary to a hydatic cyst of the liver. Its primitive form is considered due to a haematogenous diffusion through arterial vessels. We report a historical case of peritoneal hydatidosis concomitant to a hepatic localisation, in a 13-year-old girl. Diagnosis was performed by ultrasounds and CT scan, and a strong positive serology. Surgical cure has been completed by pharmacologic treatment with the aim of avoiding a relapse. A brief reminder of the therapeutic management of the disease is provided. PMID- 14550980 TI - [Systemic infantile mastocytosis: about a case with respiratory and digestive involvement]. AB - Systemic mastocytosis is rare in children and is characterized by an abnormal proliferation and infiltration of mast cells in different tissues. CASE REPORT: We report a case of systemic mastocytosis presenting cutaneous symptoms during the neonatal period. Later evolution was characterized by systemic manifestations consisting of recurrent respiratory infections with wheezing and a digestive involvement that included abdominal pain, hepatosplenomegaly and a nodular, hemorrhagic infiltrate in a low esophagus. The diagnosis was confirmed by histology and biology, notably increased histamine concentrations in blood and urines. Improvement of the respiratory and digestive symptoms was obtained with treatment by histamine H1 and H2 receptors antagonists. CONCLUSION: Respiratory manifestations and nodular infiltration of the digestive tract are rare in systemic mastocytosis. The prognosis is conditioned by complications such as malignancy and the persistence of the disease till the adult age. PMID- 14550981 TI - [Experimental ureteral obstruction and knockout animals]. AB - Obstructive uropathies caused by congenital malformations of the urinary tract are relatively frequent in newborn. These obstructive lesions are the main cause for renal disease in infancy. Most of these uropathies are treated by surgical interventions restoring the drainage function of the urinary tract. Clinically these patients are cured but the question remains wether these patients will develop renal disease in adult life, since it has been recently shown in animal models that transient, neonatal and prenatal, ureteral obstruction induces significant renal deterioration later in life. Except for angiotensin converting enzyme inhibitors that slow down the progression of renal disease, no specific drugs reducing renal fibrosis exist. Animal models of ureteral obstruction have allowed to clearly identify the events leading to tubulointerstitial fibrosis. Furthermore, more recently, the use of ureteral obstruction in genetically engineered animals has shown pro- and anti-fibrotic properties of a large number of molecules. These studies using genetically engineered animals have suggested several new future promising therapeutic directions to treat renal fibrosis. PMID- 14550982 TI - [Dental procedures in children with nephropathy]. PMID- 14550983 TI - [Does non IgE-dependent sensitization to cow's milk proteins influence chronic cough and asthma in children?]. PMID- 14550984 TI - [Screening of congenital dislocation of the hip in the newborn. A systematic and rigorous clinical examination. A restricted use of imaging]. AB - (1) Clinical examination of the hips should be systematically performed in the newborn, in order to detect a possible congenital dislocation. This translates into hip joint instability, which can be evidenced by both Ortolani's and Barlow's signs, the latter being more sensitive. Special attention should be paid for abduction range, as limited abduction is a warning sign. (2) Use of imaging, especially sonography will be restricted to specific cases only. It's prescription, technical realization as well as interpretation have to be carefully done, so that it is actually relevant to uncertain or "at risk" situations. (3) Treatment if needed, should be ideally managed by a paediatric orthopaedic surgeon. PMID- 14550985 TI - [Guidelines for diagnosis of whooping cough]. PMID- 14550986 TI - [Whooping cough: biological diagnosis]. AB - Whooping cough is a respiratory infection particularly severe for infants. The agents of the disease are the bacteria Bordetella pertussis and B. parapertussis. Before vaccination, the disease affected majority of the children. Due to the high circulation of the bacteria, adolescents and adults were regularly in contact with the bacteria and had natural boosters. Forty years after generalized vaccination in some countries, a change in the transmission of the disease is observed. Children between 2 and 5-10 years of age are not infected (depending of the vaccine used and the vaccine calendar) because they were vaccinated in their infancy. However, adolescents and adults are becoming susceptible because they do not receive any vaccinal or natural boosters. They can be infected and contaminated infants are too young to be vaccinated. Clinical symptoms in adolescents and adults, previously vaccinated or infected, are very variable and for this reason biological diagnosis are now necessary to confirm the infection. These diagnostics are culture, PCR and serology. PMID- 14550987 TI - [Respiratory function measurements in infants]. PMID- 14550991 TI - Personal perspectives. PMID- 14550993 TI - Commentary on "Discriminatory hCG zone: its use in the sonographic evaluation for ectopic pregnancy". Kadar N, DeVore G, Romero R. Discriminatory hCG zone: its use in the sonographic evaluation for ectopic pregnancy. Obstet Gynecol 1981;58:156 61. PMID- 14550995 TI - Potentially preventable excess mortality among higher-order multiples. AB - OBJECTIVE: To estimate the level of potentially preventable excess mortality achievable by avoiding the creation of higher-order multiple gestation with assisted reproductive technologies. METHODS: This was a retrospective cohort study of multiple pregnancies delivered in the United States between 1995 and 1997 involving 304,466 twins, 16,068 triplets, 1448 quadruplets, and 180 quintuplets. We used the generalized estimating equation framework to compute adjusted relative risks for combined perinatal and infant mortality (early mortality). We then calculated potentially preventable excess mortality among higher-order gestations, using twins and triplets sequentially as the referent category. RESULTS: Early mortality increased significantly with each additional fetus in a dose-dependent fashion (P <.001), corresponding to relative risks (95% confidence interval) of 2.4 (2.2, 2.6) for triplets, 3.3 (2.5, 4.4) for quadruplets, and 10.3 (5.0, 21.4) for quintuplets. The creation of twin rather than quadruplet pregnancies would be associated with a substantially higher level of preventable excess mortality (70%) than the creation of triplet pregnancies (28%). By contrast, limiting quintuplets to twins or triplets did not exhibit a similar level of difference (89% versus 75%, respectively). CONCLUSIONS: Our findings support the need for regulating the number of transferred embryos that result in quadruplet and quintuplet pregnancies. PMID- 14550996 TI - Prospective risk of fetal death in singleton, twin, and triplet gestations: implications for practice. AB - OBJECTIVE: To evaluate the prospective risk of fetal death in singleton, twin, and triplet pregnancies and to compare this risk with fetal and neonatal death rates. METHODS: We analyzed 11,061,599 singleton, 297,622 twin, and 15,375 triplet gestations drawn from the 1995-1998 National Center for Health Statistics linked birth and death files. Prospective risk of fetal death was expressed as a proportion of all fetuses still at risk at a given gestational age and compared with fetal death rate. Fetal death risk and neonatal death rates were represented graphically for singletons, twins, and triplets. RESULTS: The prospective risk of fetal death at 24 weeks was 0.28 per 1000, 0.92 per 1000, and 1.30 per 1000 for singletons, twins, and triplets, respectively. At 40 weeks, the corresponding risk was 0.57 per 1000 and 3.09 per 1000 for singletons and twins, respectively and, at 38 or more weeks, 13.18 per 1000 for triplets. Plots of gestation specific prospective risk of fetal death and neonatal mortality converged for singletons and twins at term but crossed for triplets at approximately 36 weeks' gestation. CONCLUSION: Prospective risk of fetal death is greater for triplets and twins than for singletons and greater for triplets than for twins during the third trimester. The pattern corroborates with uteroplacental insufficiency as a suspected underlying mechanism. When prospective risk of fetal death exceeds neonatal mortality risk, delivery might be indicated. When this model is used, this data set suggests that it might be reasonable to consider delivery of twins by 39 weeks and triplets by 36 weeks to improve perinatal outcome. PMID- 14550997 TI - Intrauterine head entrapment of a second twin by a uterine synechia. AB - BACKGROUND: Fetal head entrapment during delivery is a rare occurrence in modern obstetrics. We present a case of intrauterine head entrapment of a second twin by a uterine synechia diagnosed by ultrasound and subsequent complications that occurred. CASE: A woman with a twin intrauterine pregnancy presented at 19 weeks for evaluation of a shortened cervix. Ultrasound examination demonstrated fetal head entrapment of twin B by a uterine synechia, as well as intrauterine growth restriction (IUGR). The patient experienced preterm premature rupture of membranes at 24 weeks' gestation and underwent a cesarean delivery for a nonreassuring fetal heart rate tracing of twin B, with findings of a constricting band of fibrous tissue around the neck of twin B. CONCLUSION: Uterine synechia might cause intrauterine head entrapment and IUGR. PMID- 14550998 TI - Embolization of a uterine arteriovenous malformation followed by a twin pregnancy. AB - BACKGROUND: Uterine arteriovenous malformation is a lesion that can cause recurrent pregnancy loss. Very few successful pregnancies have been reported after selective embolization of an arteriovenous malformation. CASE: A 42-year old gravida 6 para 0 with a history of four first-trimester spontaneous abortions was found to have a uterine arteriovenous malformation by hysterosalpingogram, which was confirmed by magnetic resonance angiography. She underwent selective transcatheter embolization of the arteriovenous malformation. A twin gestation was later conceived via in vitro fertilization. The twins were delivered by cesarean at term. CONCLUSION: Uterine arteriovenous malformation can be successfully treated with selective embolization, with good obstetric outcome. PMID- 14550999 TI - Economics of reducing menstruation with trimonthly-cycle oral contraceptive therapy: comparison with standard-cycle regimens. AB - OBJECTIVE: To evaluate whether, and under what circumstances, trimonthly-cycle oral contraceptive pill (OCP) therapy is cost saving over standard-cycle OCP therapy for reducing the severity of menstrual-related symptoms. METHODS: We created three economic models, with each model representing a unique cost perspective (societal, private third party, and patient), to estimate the incremental annual costs associated with women taking trimonthly-cycle rather than standard-cycle OCP. Direct costs considered were costs for OCP prescriptions, female hygiene products, generic over-the-counter pain relievers and iron tablets, home pregnancy tests, and physician visits for menstrual related concerns. Indirect costs considered were lost wages due to menstruation related disability and opportunity costs of physician visits. We derived base case and sensitivity range estimates from prior literature, public use data, expert opinion, and cross-sectional survey data that we collected from 59 adult women taking standard-cycle OCP visiting a large, community-based gynecology office. All costs were adjusted to 2002 US dollars. RESULTS: Under base-case assumptions that both regimens were equivalently priced (1.16 US dollars/pill) and trimonthly OCP reduces hygiene product use by 50%, annual societal costs per person were 460 US dollars for trimonthly OCP and 501 US dollars for standard OCP (incremental cost savings = 41 US dollars). Assuming no difference in monthly drug copayment costs between the two regimens (5.00 US dollars), annual private third-party costs were nearly identical at 365 US dollars per regimen. Annual patient costs were 95 US dollars for trimonthly OCP and 136 US dollars for standard OCP. Trimonthly OCP remained cost saving to society as long its price remained below a 9% premium to standard OCP prices. The degree to which trimonthly OCP reduces hygiene product use also significantly influenced the cost difference between the two regimens. CONCLUSION: Compared with standard-cycle OCP, trimonthly-cycle OCP appears to be associated with significant societal and patient cost savings. These savings are highly conditional upon trimonthly OCP being priced similarly to standard OCP. PMID- 14551000 TI - Understanding risk: a randomized controlled trial of communicating contraceptive effectiveness. AB - OBJECTIVE: To determine which of three different approaches increased women's understanding of risk of pregnancy associated with different contraceptive methods. METHODS: We randomly assigned 461 reproductive-age women to one of three tables presenting pregnancy risk (Food and Drug Administration table with numbers, World Health Organization table with numbers and categories, or table with categories). We evaluated participant knowledge before and after being shown the assigned table. RESULTS: The most important reason for choosing a contraceptive was how well it works (53%), followed by ease of use (13%), and protection against sexually transmitted disease or human immunodeficiency virus (11%). Before looking at the tables, about half the participants knew that hormone shots are more effective than pills (48%) and that pills are more effective than condoms (57%). For these two key comparisons, the category table compared to the Food and Drug Administration table with numbers improved knowledge significantly more (37% versus 20% and 27% versus 14%; both P <.05). Compared with those assigned to the Food and Drug Administration table with numbers, significantly fewer participants assigned to the category table said the table was difficult to read (6% versus 19%; P <.01). Most participants in all three groups said their assigned table provided enough information to choose a contraceptive method. CONCLUSION: The table with categories communicated relative contraceptive effectiveness better than the tables with numbers. However, without being presented with numbers, participants grossly overestimated the absolute risk of pregnancy using contraceptives. A combination of categories and a general range of risk for each category may provide the most accurate understanding of both relative and absolute pregnancy risk. PMID- 14551001 TI - Outcome of fertility-sparing treatment with progestins in young patients with endometrial cancer. AB - OBJECTIVE: To evaluate the safety of fertility-sparing hormonal therapy for endometrial cancer in young patients. METHODS: We reviewed the clinical and pathologic records of patients diagnosed with endometrial adenocarcinoma before the age of 40, who were treated and followed over a 30-year period in the Division of Gynecologic Oncology. All patients who underwent conservative management with progestins (n = 13) are the subjects of this study. RESULTS: Follow-up was available for all 13 patients, with a mean follow-up of 82 months. All patients responded to treatment within a mean period of 3.5 months, with normal pathology on follow-up endometrial samplings. Six patients had a recurrence within a period extending between 19 and 358 months (median 40 months). Four patients were treated with a second course of progestins, and all had a histologic complete response. As of the time of preparation of this report, nine healthy infants had been born, and all the patients remained without evidence of disease. CONCLUSION: Conservative management of well-differentiated endometrial carcinoma in young patients, combined with assisted reproductive technologies, if needed, does not seem to worsen the prognosis. This approach also provides the possibility of conceiving and carrying a normal pregnancy. PMID- 14551002 TI - Superiority of electrocautery over the suture method for achieving cervical cone bed hemostasis. AB - OBJECTIVE: To compare the efficacy of electrocautery with that of the suture method for achieving hemostasis of the cervical cone bed. METHODS: We performed a retrospective chart review of all patients who underwent cold-knife conization of the cervix over a 5-year period. Patients were categorized into two groups: the cautery group, in which the cone bed was electrocauterized with a hand-held electrocoagulation device; and the suture group, in which hemostasis was achieved by a continuous locking suture placed circumferentially around the cone bed. Outcome measures evaluated include estimated blood loss, operative time, and incidence of complications, including secondary hemorrhage, cervicitis, and cervical stenosis. Data were analyzed by Student t test, chi(2) test, linear regression, and multiple logistic regression where appropriate. RESULTS: There were 156 women in the cautery group and 35 in the suture group. The cautery group had significantly lower estimated blood loss (27 mL versus 101 mL; P <.01) and shorter operative time (34 versus 43 minutes; P <.01) than the suture group. The procedure-related complication rate was 6.4% in the cautery group, compared with 14.3% in the suture group (P = nonsignificant). A higher use of lateral sutures, vasopressors, and thrombotic agents was seen in the cautery group. However, even after adjusting for these variables, mean estimated blood loss (33 mL, P <.01) and mean operative time (34 minutes, P <.01) were significantly less in the cautery group than in the suture group. CONCLUSIONS: Cauterization of the cone bed is superior to suture as a method of achieving hemostasis, with significantly less blood loss and shorter operative time. PMID- 14551003 TI - Intrapartum computerized fetal heart rate parameters and metabolic acidosis at birth. AB - OBJECTIVE: To estimate to what extent computerized fetal heart rate (FHR) parameters are affected by labor and to estimate the relationship between FHR parameters and the degree of fetal metabolic acidosis in laboring patients at term. METHODS: Fifty-one women between 37 and 42 weeks' gestational age were recruited prospectively in the following groups: 1) nonlaboring women, and 2) laboring women requiring fetal scalp electrode for continuous electronic FHR monitoring. Computerized FHR analysis was performed for 1 hour within 6 hours of delivery in the nonlaboring group and continuously throughout labor in the laboring group. Multiple linear regression analysis was used to determine the relationship between individual FHR parameters during the last hour before delivery and the degree of metabolic acidosis at birth. RESULTS: The umbilical cord artery base excess and pH did not show any significant correlation with any of the computer-derived FHR parameters studied. Both umbilical cord venous base excess and pH were inversely related to the number of large FHR decelerations (r = -.46, P <.01 and r = -.56, P <.01, respectively). Labor was associated with a 31% increase in both short- and long-term FHR variation in the reassuring FHR tracing group when compared with nonlaboring women. Although this increase in FHR variation was not seen in the nonreassuring FHR tracing group, there was no relationship to the degree of metabolic acidosis at birth. CONCLUSION: In term pregnant women with reassuring FHR tracing, labor causes an increase in both short- and long-term FHR variation, which was abolished in the presence of nonreassuring FHR tracing. Computer-derived FHR parameters studied during the last hour of labor were not correlated with the degree of metabolic acidosis as measured in the umbilical artery at birth. PMID- 14551004 TI - Asthma symptoms, severity, and drug therapy: a prospective study of effects on 2205 pregnancies. AB - OBJECTIVE: To prospectively examine in pregnant women whether asthma or asthma therapy influenced preterm delivery, intrauterine grown restriction (IUGR), or birthweight. METHODS: We enrolled 873 pregnant women with a history of asthma, of whom 778 experienced asthma symptoms or took medication, and 1333 women with no asthma history, including 884 women with neither asthma diagnosis nor symptoms and 449 with symptoms but no diagnosis. Asthma symptoms, medication, and severity were classified according to 2002 Global Initiative for Asthma guidelines. RESULTS: Preterm delivery was not associated with asthma diagnosis, severity, or symptoms but was associated with use of controller medications, independent of symptoms, specifically oral steroids and theophylline. Gestation was reduced by 2.22 weeks in women using oral steroids daily (P =.001) and 1.11 weeks after theophylline (P =.002). We observed a 24% (5-47%) increased risk for IUGR with each increased symptom step, which increased further in symptomatic women with no asthma diagnosis (31%, 4-65%) compared with women with neither asthma nor symptoms. CONCLUSIONS: We found no effect of asthma symptoms or severity on preterm delivery but observed increased risks associated with use of oral steroid and theophylline. Intrauterine growth restriction was associated with asthma severity, which possibly reflects a hypoxic fetal effect. Women with asthma symptoms but no diagnosis were at particular risk of undermedication and delivering IUGR infants. These observations support guidelines that advocate active management of pregnant patients with mild or moderate asthma with beta(2) agonists, with oral steroids added only if severity increases. Symptomatic patients without an asthma diagnosis might need to be equally managed. PMID- 14551005 TI - Prenatal screening for infectious diseases and opportunities for prevention. AB - OBJECTIVE: To characterize adherence with recommendations for prenatal infectious disease screening and missed opportunities for prevention of congenital and perinatal infections. METHODS: Demographic, prenatal, and peripartum information was abstracted from labor and delivery records of a random, stratified sample of live births in 1998 and 1999 to residents of eight active surveillance areas. Adherence with prenatal screening recommendations was evaluated for hepatitis B, syphilis, rubella, human immunodeficiency virus (HIV), and group B streptococcus (GBS). Characteristics of missed opportunities for disease prevention were assessed by univariate and multivariable analysis to account for survey design. RESULTS: Prenatal screening rates for hepatitis B surface antigen (HBsAg) (96.5%), syphilis (98.2%), and rubella (97.3%) were high. Areas of excess syphilis morbidity did not adhere to recommendations for third-trimester retesting. Testing rates for HIV (57.2%) and GBS (52.0%) were lower and had wide geographic variation. Postpartum rubella vaccination was documented for only 65.7% of rubella-susceptible women. Inadequate prenatal care was the single strongest predictor of missed opportunities for prenatal testing (relative risk 14.6; 95% confidence interval 6.3, 33.7). Blacks were less likely than whites to receive adequate prenatal care and prenatal tests, more likely to test positive for HBsAg and syphilis, and less likely to receive recommended prevention interventions such as postpartum rubella vaccination for susceptible women. CONCLUSIONS: Adherence to both long-standing and more recent recommendations for congenital and perinatal disease prevention can be improved, thus perhaps reducing racial disparities in the use of prenatal screening and appropriate prevention interventions. PMID- 14551006 TI - Rehydration of air-dried cervical smears: a feasible alternative to conventional wet fixation. AB - OBJECTIVE: To evaluate the effect of rehydration of air-dried cervical smears on staining quality and to find out whether rehydration techniques can be adopted as an alternative method, especially in high-volume, resource-limited settings. METHODS: A total of 950 paired wet-fixed and air-dried-rehydrated cervical smears collected by paramedical workers from an urban slum of Delhi, India, were compared for staining quality by assessing different cytoplasmic and nuclear parameters. RESULTS: The staining quality in air-dried-rehydrated smears was satisfactory and comparable to wet-fixed smears (61.3% versus 63.6%). The staining was superior in rehydrated smears in 26.5%, as compared with 15.4% in wet-fixed ones (P <.001). Unsatisfactory staining was observed in 12.2% of rehydrated smears, as compared with 21% of wet-fixed smears. The smear background was cleaner in rehydrated smears. Red blood cells were observed in only 3% of rehydrated smears, in comparison to 12% in wet-fixed ones (P <.001). CONCLUSION: Rehydration of air-dried cervical smears is a suitable alternative to wet-fixed smears. The staining quality is either the same as or better than wet-fixed smears, and the unsatisfactory rate is lower. This technique is simple and can be conveniently adopted in resource-limited settings. PMID- 14551007 TI - Changes in cervical cancer incidence after three decades of screening US women less than 30 years old. AB - OBJECTIVE: To examine incidence trends of invasive cervical carcinoma in US women less than 30 years old from 1973 to 1999 by histologic type. METHODS: Incidence rates were obtained from the National Cancer Institute's Surveillance, Epidemiology and End Results (SEER) database and stratified by age and three histologic categories: all histologic types, squamous cell carcinoma, and adenocarcinoma. The estimated annual percent change was calculated by joinpoint regression to measure trends during the entire evaluation period and a recent subperiod (1985-1999). All statistical tests were two-sided. RESULTS: Incidence rates of cervical carcinoma overall and squamous cell carcinoma specifically declined during 1973-1999, with estimated annual percent changes of -0.94% (95% confidence interval [CI] -1.47%, -0.41%) and -1.10% (95% CI -1.59%, -0.62%), respectively. Rates of adenocarcinoma increased (+2.90%; 95% CI 1.34%, 4.49%), though trends have been stable since 1990. All cancers were uncommon: the annual incidence per million women ranged from 8.0 to 14.3 for squamous cell carcinoma and from 0.7 to 2.7 for adenocarcinoma. For women younger than 20 years, no statistically significant changes were observed, but cancers were rare (0-3 per million women annually). CONCLUSIONS: More investigation is necessary to clarify the contribution of screening to declines in the squamous cell carcinoma rate and to determine the etiology of adenocarcinoma rate increases over the last three decades in US women less than 30 years old. Because of the small number of actual observed cases, caution must be exercised in interpreting these trends. PMID- 14551008 TI - Cervical cancer and microchimerism. AB - OBJECTIVE: To determine whether microchimerism is involved in the pathogenesis or progression of cervical cancer. METHODS: Cervical tissue was obtained from eight women who had at least one live-born son and who underwent radical hysterectomy after a diagnosis of cervical cancer. Control tissue was obtained from four women without cervical cancer who had at least one live-born son and from three women with cervical cancer and no male births. Tissue sections were analyzed with fluorescence in situ hybridization for the presence of fetal cells, defined by an X and Y chromosome. Immunolabeling was used to determine the phenotype of the presumed fetal cells. RESULTS: Male cells were found in cervical tissue from all four patients for whom large sections (approximately 1.5 x 2 cm) were analyzed. Only one male cell was found in two of the four patients for whom small biopsy specimens (approximately 0.1 x 0.5 cm) were analyzed. No male cells were found in tissue specimens from controls, whether they were small or large sections. In immunolabeling studies, eight of 18 male cells from one patient were CD45 positive and nine of 37 male cells from two patients were cytokeratin-positive. No cells were positive for both markers. CONCLUSION: Cervical cancer might be associated with microchimerism, possibly from fetomaternal cell trafficking. These results further expand the potential relationship between microchimerism and disease in women. PMID- 14551009 TI - Human immunodeficiency virus retesting during pregnancy: costs and effectiveness in preventing perinatal transmission. AB - OBJECTIVE: To estimate the incremental societal costs and effectiveness of a second human immunodeficiency virus (HIV) antibody test during the third trimester of pregnancy compared with no second test. METHODS: We used a decision tree in this cost-effectiveness analysis to model outcomes among pregnant women in high-risk communities and nationwide who received an initial, negative HIV antibody test during the first trimester. The main outcome measure was discounted costs per year of infant life saved. RESULTS: In high-risk communities with estimated HIV incidence of 6.2 per 1000 person-years, a second HIV test compared with no second test would detect 192 infections in women, prevent approximately 37 infant infections, and save 655 infant life-years per 100,000 women tested. Net savings would be 5.2 million US dollars. Applied to an estimated national incidence of.17 per 1000 person-years, a second test would detect 5.3 infections in women, prevent 1.3 infant infections, and save 23.3 infant life-years per 100,000 women tested. Net costs would be 1.06 million US dollars, or 45,708 US dollars for each year of infant life saved. A second test would result in net savings in populations with HIV incidence of 1.2 per 1000 person-years or higher. CONCLUSION: Health care providers serving women in communities with an HIV incidence of 1 per 1000 person-years or higher should strongly consider implementing a second voluntary universal HIV test during the third trimester. Providers serving lower-risk communities should pilot second testing to assess community-specific costs. PMID- 14551010 TI - Changes in maternal characteristics and obstetric practice and recent increases in primary cesarean delivery. AB - OBJECTIVE: To estimate the contribution of changes in maternal characteristics (namely, age, parity, prepregnancy weight, weight gain in pregnancy, smoking status) and obstetric practice (namely, labor induction, epidural anesthesia, delivery by an obstetrician, midpelvic forceps delivery) to recent increases in primary cesarean delivery rates. METHODS: We studied all deliveries in Nova Scotia, Canada, between 1988 and 2000 after excluding women who had a previous cesarean delivery (n = 127,564). Logistic regression was used to study the effect of changes in maternal characteristics and obstetric practice on primary cesarean delivery rates. The effect of changes in midpelvic forceps delivery was examined through ecologic Poisson regression. RESULTS: Primary cesarean delivery rates increased from 13.4% of deliveries in 1988 to 17.5% in 2000. This was due to increases in cesarean deliveries for dystocia (14% increase), breech (24% increase), suspected fetal distress (21% increase), hypertension (47% increase), and miscellaneous indications (73% increase). Adjustment for maternal characteristics reduced the temporal increase in primary cesarean delivery rates between 1988-1991 and 1998-2000 from 21% (95% confidence interval [CI] 16%, 25%) to 2% (95% CI -2%, 7%). Additional adjustment for obstetric practice factors further reduced period effects. Midpelvic forceps delivery was significantly and negatively associated with primary cesarean delivery (P =.001). CONCLUSION: Recent increases in primary cesarean delivery rates are a consequence of changes in maternal characteristics. Obstetric practice, which has altered due to changes in maternal characteristics and concerns related to fetal and maternal safety, has also contributed to increases in primary cesarean delivery. PMID- 14551011 TI - Clinical outcome of fetuses with sonographic diagnosis of isolated micrognathia. AB - OBJECTIVE: To describe the clinical outcome of fetuses with the prenatal sonographic diagnosis of isolated micrognathia. METHODS: A retrospective review of fetuses and infants with the prenatal diagnosis of isolated micrognathia for April 1990 to August 2001 was undertaken. Isolated micrognathia was considered if no other anatomic, growth, or amniotic fluid abnormalities were detected by a detailed ultrasound examination. Sources of outcome data included maternal and neonatal medical records, prenatal genetics records, and karyotype results. RESULTS: Fifty-eight fetuses with the diagnosis of micrognathia were identified. Fifteen fetuses (26%) had isolated micrognathia by prenatal sonogram. After neonatal examination, 14 of 15 were found to have at least one additional abnormality. Eleven had a cleft of the soft and/or hard palate. Seven (54%) of 13 live-born neonates had mild to severe airway obstruction that required intervention. Four (31%) of 13 experienced feeding difficulties of varying duration. Follow-up data were available for 1 to 10 years. Eight (62%) of 13 children are reported to be doing well. Five (38%) of 13 children are reported to have mild to severe developmental delay. CONCLUSION: If micrognathia is the only sonographic finding identified, physicians and families should be prepared for possible respiratory difficulty at delivery, the presence of a cleft palate, and/or developmental delay. PMID- 14551012 TI - Maternal cardiac function in twin pregnancy. AB - OBJECTIVE: To investigate maternal cardiac function in twin pregnancy. METHODS: We conducted a cross-sectional study of 119 pregnant women with twin pregnancies at 10-40 weeks' gestation. Two-dimensional and M-mode echocardiography of the left ventricle was performed in the left lateral decubitus position to assess left ventricular longitudinal and transverse systolic function. The measurements were compared with those obtained from 128 women with singleton pregnancies previously reported. RESULTS: In twin pregnancies, compared with singletons, maternal cardiac output was greater by 20% (P <.001), because of a greater stroke volume (15%, P <.001) and heart rate (3.5%; P =.04). Furthermore, in women with twins there were greater left ventricular end-diastolic and left ventricular end systolic dimensions, left ventricular mass (13.5%; P <.001), fractional shortening (3%; P =.04), and ejection fraction (2.5%; P =.04). Mean arterial pressure and global time to shortening in women with twins, compared with singletons, were less in the first trimester by approximately 2%, but after midpregnancy they increased progressively, so that at term the measurements were greater by 3% and 5.7%, respectively (P =.03). Conversely, long axis shortening in women with twins, compared with singletons, was greater in the first trimester by approximately 6.5%, but at term it was 3% less (P =.01). Twin pregnancies that subsequently developed preeclampsia had a hemodynamic profile similar to the rest of the twin population. CONCLUSION: Twin pregnancy is characterized by an even more hyperdynamic circulation than singleton pregnancy. Left ventricle longitudinal systolic function and mean arterial pressure are more abruptly affected after 20 weeks compared with singleton pregnancies. PMID- 14551013 TI - Quantification of ovarian power Doppler signal with three-dimensional ultrasonography to predict response during in vitro fertilization. AB - OBJECTIVE: To evaluate whether power Doppler predicts ovarian response to gonadotrophin stimulation during in vitro fertilization (IVF). METHODS: Forty five women were divided into low-reserve (n = 12) and normal-reserve (n = 33) ovarian groups, according to antral follicle count. Transvaginal three dimensional power Doppler ultrasonographic examinations were performed after pituitary downregulation and after gonadotrophin stimulation. The antral follicle count, ovarian volume, vascularization index, flow index, vascularization flow index, and mean gray value were measured and related to the number of oocytes retrieved and the pregnancy rate. RESULTS: The number of oocytes retrieved correlated with the antral follicle count (R =.458, P =.004) and ovarian volume (R =.388, P <.016) but not with vascularization index, flow index, vascularization flow index, or mean gray value after pituitary suppression. There was an increase in vascularization index (P <.017), flow index (P <.001), and vascularization flow index (P <.007) during gonadotrophin stimulation in the normal-ovary group but not in the low-ovarian-reserve group. CONCLUSION: According to our results, quantification of power Doppler signal in the ovaries after pituitary suppression does not provide any additional information to predict the subsequent response to gonadotrophin stimulation during IVF. The increase in ovarian power Doppler signal during gonadotrophin stimulation is related to the antral follicle count observed after pituitary suppression. PMID- 14551014 TI - Efficacy and tolerability of a novel estradiol vaginal ring for relief of menopausal symptoms. AB - OBJECTIVE: To assess the efficacy, tolerability, and acceptance of a vaginal ring delivering the equivalent of 50 or 100 microg per day of estradiol (E2), compared with placebo, for relief of moderate to severe vasomotor symptoms and urogenital symptoms in postmenopausal women. METHODS: Women with moderate to severe vasomotor symptoms (seven or more per day or 56 per week average) received 13 weeks of treatment with a vaginal ring delivering 50 microg per day E2 (n = 113) or 100 microg per day E2 (n = 112), or a placebo vaginal ring (n = 108). Severity of vasomotor symptoms was assessed by a daily diary card and the Greene Climacteric Scale. Urogenital signs and symptoms were evaluated via patient and physician assessment and vaginal cytology. Participant satisfaction with the vaginal ring was evaluated via questionnaire. RESULTS: Vasomotor symptoms significantly improved in both treatment groups, compared with placebo (P <.05). There was a trend toward greater improvement in patient assessment of urogenital signs with active rings compared with placebo. For women with vaginal atrophy at baseline (n = 60), the maturation index improved significantly in both treatment groups compared with placebo. Total Greene Climacteric Scale scores significantly improved for both E2 vaginal ring groups (P <.05) compared with placebo. The vaginal rings were well tolerated. Most adverse events were mild or moderate and consistent with estrogen therapy. CONCLUSION: A novel vaginal ring delivering the equivalent of 50 or 100 microg per day of E2 significantly reduced the number and severity of vasomotor symptoms and improved urogenital symptoms, compared with placebo. The E2 vaginal ring was well tolerated. PMID- 14551015 TI - Pain-free efficacy after treatment with sumatriptan in the mild pain phase of menstrually associated migraine. AB - OBJECTIVE: To estimate the efficacy of sumatriptan 50-mg and 100-mg tablets in menstrually associated migraine when treatment is administered during the mild pain phase. METHODS: A randomized, double-blind, placebo-controlled, single attack study was conducted. Menstrually associated migraine was defined as any migraine beginning on or between day -2 and day 4, with day 1 = first day of flow. Patients had at least a 1-year history of migraine as defined by International Headache Society criteria and reported regularly occurring menstrually associated migraines typically having a mild pain phase. Patients treated attacks within 1 hour of the onset of pain but only if the pain was mild at onset and while the pain was still mild. RESULTS: In the 349 women with menstrually associated migraine, sumatriptan was significantly more effective than placebo: 61% and 51% of patients who used sumatriptan 100 mg and 50 mg, respectively, were pain-free 2 hours after treatment compared with 29% of patients who used placebo (P <.001 for both comparisons). At 2 hours, 51% and 45% of patients who used sumatriptan 100 mg and 50 mg were free of pain and associated symptoms (photophobia, phonophobia, nausea, vomiting) compared with 25% of placebo patients (P <.001 for both comparisons). Adverse events were low for sumatriptan 100 and 50 mg, and both doses were generally well tolerated. CONCLUSION: Sumatriptan 50-mg and 100-mg tablets are generally well tolerated and effective in providing pain-free relief and relief of the associated symptoms of menstrually associated migraine when administered in the mild pain phase. PMID- 14551016 TI - Successful medical treatment with danazol after failed embolization of uterine arteriovenous malformation. AB - BACKGROUND: Transcatheter arterial embolization has been the therapy of choice for uterine arteriovenous malformations, whereas medical therapy has not been popular because of patient propensity to bleed. CASE: A 29-year-old woman, gravida 3, para 0, was diagnosed with uterine arteriovenous malformation. Because initial treatment with uterine artery embolization was unsuccessful, she was ultimately treated with danazol. Resolution of the lesion after 2 weeks of danazol therapy was observed. As of follow-up at 16 months, she has remained free from further abnormal bleeding episodes and recurrence of the lesion. CONCLUSION: Danazol has the potential for medical management of uterine arteriovenous malformations in hemodynamically stable patients who do not respond to embolization. PMID- 14551017 TI - Helicobacter pylori infection and dyspepsia in pregnancy. AB - OBJECTIVE: To investigate the relationship of Helicobacter pylori infection with dyspeptic symptoms in early and late pregnancy. Infection with H pylori and pregnancy outcome were also assessed. METHODS: H pylori seropositivity was determined in 416 antenatal patients at 10 to 14 weeks' gestation. Dyspeptic symptoms were recorded at 10 to 14 weeks' gestation and at 30 to 32 weeks' gestation by means of a well-validated questionnaire. Details of pregnancy outcome were recorded from patients' case notes. RESULTS: The incidence of H pylori infection in our population was 41.8% (164 of 404). Patients infected with H pylori were no more likely than controls to experience dyspepsia at 10 to 14 weeks' or at 30 to 32 weeks' gestation (P =.75 and.43, respectively). CONCLUSION: H pylori infection was not associated with preterm delivery, non reassuring fetal status in labor, or birth weight less than the 10th centile (P =.17,.57, and.19, respectively).H pylori infection is not associated with an increase in dyspepsia or with maternal or neonatal morbidity. PMID- 14551018 TI - Gestational diabetes mellitus and lesser degrees of pregnancy hyperglycemia: association with increased risk of spontaneous preterm birth. AB - OBJECTIVE: To investigate whether different degrees of maternal glucose intolerance are associated with the risk of spontaneous preterm birth. METHODS: We performed a cohort study of 46,230 pregnancies screened by a 50-g, 1-hour oral glucose tolerance test between 24 and 28 gestation weeks at the Northern California Kaiser Permanente Medical Care Program. Spontaneous preterm birth was defined as an infant born at less than 37 gestation weeks with at least one of the following: spontaneous labor, preterm premature rupture of membranes, or incompetent cervix. Glucose tolerance status was categorized as normal screening (1-hour plasma glucose less than 140 mg/dL), abnormal screening (1-hour plasma glucose of at least 140 mg/dL with a normal diagnostic 100-g, 3-hour oral glucose tolerance test result), Carpenter-Coustan (plasma glucose measurements during the diagnostic oral glucose tolerance test met the thresholds but were lower than the National Diabetes Data Group thresholds), and gestational diabetes mellitus (GDM) by the National Diabetes Data Group criteria. RESULTS: One thousand nine hundred fifty-six spontaneous preterm births occurred. Age-adjusted incidences of spontaneous preterm birth were 4.0% in normal screening, 5.0% in abnormal screening, 6.7% in Carpenter-Coustan, and 6.7% in GDM. In a logistic regression model adjusted for age, race-ethnicity, preeclampsia-eclampsia-pregnancy-induced hypertension, chronic hypertension, polyhydramnios, and birth weight for gestational age, pregnancies with abnormal screening, Carpenter-Coustan, and GDM had a significantly higher risk of spontaneous preterm birth than pregnancies with normal screening (relative risk [95% confidence interval]: 1.23 [1.08, 1.41], 1.53 [1.16, 2.03], and 1.42 [1.15-1.77], respectively). CONCLUSION: The risk of spontaneous preterm birth increased with increasing levels of pregnancy glycemia. This association was independent of perinatal complications that could have triggered early delivery. PMID- 14551019 TI - Management of diabetes mellitus complicating pregnancy. AB - Diabetes mellitus complicates 3-5% of all pregnancies and is a major cause of perinatal morbidity and mortality, as well as maternal morbidity. The availability of a variety of new insulins, the insulin pump, and self-monitoring of blood glucose have revolutionized the care of the pregnancy complicated by diabetes mellitus. However, challenges remain in caring for the pregnant patient with pregestational diabetes. Relatively few women receive preconceptional counseling, and major fetal malformations as a result of poor glucose control before and during the early weeks of gestation have emerged as the major cause of perinatal mortality. When the patient has diabetic vasculopathy, the obstetrician, maternal-fetal specialist, and/or endocrinologist and other members of the health care team must perform a challenging balancing act that promotes fetal health while minimizing maternal risk. As obesity increases in this country and our population becomes more diversified, the rate of gestational diabetes mellitus (GDM) will rise. Although there is controversy regarding which diagnostic standards to use for GDM, there is agreement that excellent blood glucose control, with diet and, when necessary, insulin will result in improved perinatal outcome. Finally, the goal of our educational programs should be not only to improve pregnancy outcome but also to promote healthy lifestyle changes for the mother that will last long after delivery. PMID- 14551021 TI - Antiphospholipid syndrome: obstetric diagnosis, management, and controversies. PMID- 14551023 TI - ACOG practice bulletin number 47, October 2003: Prophylactic Antibiotics in Labor and Delivery. AB - The use of antibiotics to prevent infections during pregnancy and the puerperium is clearly different from the use of antibiotics to treat established infections. Prophylactic antibiotics are frequently prescribed on an "empiric" basis, which for many years was thought to have few adverse consequences. Concerns about the emergence of resistant strains of common bacteria, in addition to the emergence of strains with increased virulence, have resulted in increased scrutiny of the use of antibiotics, particularly in the hospital setting. Cost considerations also affect the use and choice of prophylactic agents. The purpose of this Practice Bulletin is to present a review of clinical situations in which prophylactic antibiotics are frequently prescribed and to weigh the evidence supporting the use of antibiotics in these scenarios. PMID- 14551024 TI - ACOG committee opinion number 286, October 2003: patient safety in obstetrics and gynecology. AB - Emphasis on patient safety has increased in the past few years mostly in response to the Institute of Medicine report "To Err Is Human: Building a Safer Health System." Obstetrician*gynecologists should incorporate elements of patient safety into their practices and also encourage others to use these practices. The American College of Obstetricians and Gynecologists (ACOG) is committed to improving quality and safety in women's health care. The Institute of Medicine report, "To Err Is Human: Building a Safer Health System," notes that errors in health care are a significant cause of death and injury. Despite disagreements over the actual numbers cited, all health care professionals agree that patient safety is extremely important and should be addressed by the overall health care system. The American College of Obstetricians and Gynecologists continues to emphasize its long-standing commitment to quality and patient safety by codifying a set of objectives that should be adopted by obstetrician*gynecologists in their practices. Obstetrician*gynecologists are encouraged to promulgate these principles in the hospitals and other settings where they practice. PMID- 14551025 TI - ACOG committee opinion number 287, October 2003: newborn screening. AB - Newborn screening programs have enormous public health benefits and have been effective in identifying newborns who can benefit from early treatment. Because of advances in genetics and technology, newborn screening programs are capable of testing for more than 30 disorders, including infections, genetic diseases, and inherited and metabolic disorders. Many important issues surround the debate on universal screening, including financial resources, level of screening, continuity of care, and informed consent. To date, policy on newborn screening has been fragmented; however, efforts are underway to ensure uniformity and equity for all newborns. Obstetrician-gynecologists can continue to work to improve the health of their patients by informing expectant families of the newborn screening process. PMID- 14551026 TI - ACOG committee opinion number 288, October 2003: professional liability and gynecology-only practice. AB - Some Fellows of the American College of Obstetricians and Gynecologists limit the scope of their practices solely to gynecology. The College considers early pregnancy complications (often up to 12-14 weeks of gestation) to be within the definition of gynecology. Liability insurance should cover the role of gynecologists in the management of early pregnancy-related conditions. PMID- 14551027 TI - CTLA4-Ig abrogates the anti-globulin response and prolongs cardiac allograft survival after anti-CD2 treatment. AB - CD2 is expressed on T cells and NK cells and is important in T cell activation, making it a potential target for immune intervention. Here, we report a series of experiments aimed at defining the ability of mAbs directed against the CD2 molecule to prevent cardiac allograft rejection in low and high responder rat strain combinations. Administration of the mouse anti-rat CD2 mAbs OX34 or OX55 around the time of transplantation prolonged survival of fully allogeneic Lewis (RT1l) cardiac allografts in low responder DA (RT1a) recipients (MST 14 days for OX55 and >100 days for OX34). Treatment with OX34 prolonged graft survival in the reciprocal high responder DA to Lewis rat strain combination (MST 19 days) and when combined with CTLA4-Ig resulted in long-term graft survival (MST>100 days). Despite these in vivo effects, OX34 had little effect on in vitro assays of lymphocyte activation. Instead, the ability of OX34 to extend allograft survival correlated with T cell depletion. Administration of OX34 induced a similar degree of CD4 T cell depletion in DA and Lewis recipients, but the CD4 depletion observed was more transient in Lewis recipients. Lewis, but not DA strain rats, developed an anti-murine Ig response. Combined treatment with CTLA4-Ig abolished the anti-globulin response to OX34 in Lewis recipients, prolonged circulation of OX34 and increased the extent and duration of CD4 depletion. We conclude that anti-CD2 treatment effectively prolongs cardiac allograft survival and addition of CTLA4-Ig increases its efficacy by abrogating the production of neutralising antibodies. PMID- 14551028 TI - NOD/SCID mice engrafted with human peripheral blood lymphocytes can be a model for investigating B cells responding to blood group A carbohydrate determinant. AB - Human antibodies (Abs) against blood group A or B carbohydrate determinant are a major barrier to ABO-incompatible organ transplantation; however, the phenotype and other properties of B cell types responding to A or B carbohydrate epitopes have not been defined. Studies here, which use fluorescein-labeled synthetic A determinant (GalNAcalpha1-3Fucalpha1-2Gal), demonstrate that B cells bearing surface IgM (sIgM) receptors recognizing blood group A carbohydrate determinant are found exclusively in a small B cell subpopulation, i.e. sIgM+ CD11b+ CD5+ B1 cells, in blood group O human peripheral blood mononuclear cells (PBMC). In order to test anti-A Abs producing capacity of the human PBMC, nonobese diabetic (NOD)/severe combined immune-deficient (SCID) mice that have been treated with rabbit anti-asialo GM1 serum to deplete natural killer cells and with 3 Gy of whole body irradiation were engrafted with blood group O or A human PBMC, followed by sensitization of human blood group A red blood cells. Anti-A-specific human Abs were detected in the sera of the mice that received blood group O human PBMC, whereas they were not detected in the sera of the mice that received blood group A human PBMC, indicating profound tolerance of auto-reactive B cells. The human PBMC-NOD/SCID chimera developed by injection of blood group O human PBMC might be a useful in vivo model to test effects of immunosuppressants or other approaches on human B cells that respond to blood group A antigens. PMID- 14551029 TI - Anti-CD20 treatment depletes B-cells in blood and lymphatic tissue of cynomolgus monkeys. AB - INTRODUCTION: Macaque species offer a valuable model for translational allo transplantation and tolerance studies. Cardiac allograft vasculopathy in Macaca fascicularis is associated with elaboration of anti-donor antibodies. Since T independent pathways of B cell activation have been described, and anti-B cell strategies have proven to be a fruitful tolerogenic adjunct in rodent and xenogenic models, here we investigate whether an anti-CD20 antibody (rituximab) would be useful to deplete B-cells in a pre-clinical allo-transplantation setting in macaques. METHODS: Three cynomolgus macaques which had previously rejected a cardiac allograft and one with concurrent subacute vascular rejection were treated weekly with rituximab 20 mg/kg i.v. for 4 and 2 weeks, respectively. B cell levels (CD19+ cells) were measured by flow cytometry in peripheral blood, spleen, lymph node and bone marrow cells at various intervals after initiation of treatment. B-cells and plasma cells were also analyzed by immunohistochemistry at necropsy in spleen, lymph node, tonsil and thymus tissue sections. Anti-donor antibody titers were measured by flow cytometry. RESULTS: B-cells expressing CD19 were not detectable in the peripheral blood in any animal within 24 h after initial treatment, or over the ensuing month. At necropsy, the germinal centers in spleen and lymph node were completely depleted of CD20+ B-cells in 2 animals, leaving a hypocellular trabecular pattern around preserved plasma cell follicles. Substantial but incomplete depletion of B-cells was demonstrated in the other 2 animals, in each instance immunohistochemical findings in spleen and lymph node exhibiting higher sensitivity for residual B-cells compared to FACS. Anti-donor antibody titers exhibited kinetics similar to untreated animals over this short follow-up. COMMENT: Treatment with anti-CD20 very efficiently depletes peripheral and tissue B-cells but not plasma cells in this macaque species. Biopsy of lymph node is necessary and may be sufficient to assess B-cell clearance in secondary lymphoid organs in this model. PMID- 14551030 TI - Early onset of rejection in concordant hamster xeno hearts display signs of necrosis, but not apoptosis, correlating to the phosphocreatine concentration. AB - BACKGROUND: The importance of apoptosis contra necrosis for ischemia/reperfusion (RP) and acute rejection in concordant rodent xenotransplantation is largely unknown. We explored this question by comparing rodent allo and concordant xenotransplants with different morphological methods to detect apoptosis and biochemical data on the levels of high-energy phosphates obtained with in vitro 31Phosphorous Magnetic Resonance Spectroscopy (31P MRS). More specifically, we applied a hitherto unused method in transplantation research, apoptosis specific biotin labeled oligonucleotides designed with a 10 base pair stem region and a 20 nucleotides large loop that form a hairpin like shape. The results obtained with this method were compared to results obtained with the more widely used in situ 3'-end labeling of DNA (TUNEL) assay and extraction and gel electrophoresis of labeled DNA (DNA laddering). METHODS: Cervical heart transplantations were performed between inbred Lewis (L) (RT1l) to L, L to DA (RT1a) rats, hamster (H) to H and H to L (X) (n=5 for all groups except for X, n=9). All hearts were subjected to 30 min of cold ischemia (+4 degrees C) and 6 h of RP before explantation. In vitro 31P MRS was used to determine the phosphocreatine (PCr), beta-adenosine triphosphate (beta-ATP) concentrations and the PCr/beta-ATP ratio of the transplants. We correlated the biochemical data to haematoxylin and eosin (H & E) stained tissue slides scored for rejection, infiltration of antibodies and complement depositions, DNA extraction and gel electrophoresis of labeled DNA (DNA laddering), in situ 3'-end labeling of DNA (TUNEL) and the apoptosis specific hairpin probe assays scoring. RESULTS: The rejection score of the xeno grafts differed significantly compared to their syngeneic hamster to hamster controls (2.40 +/- 0.25 vs. 1.20 +/- 0.20; P=0.005) and they had a significantly higher TUNEL score, 228 +/- 15 vs. 2.44 +/- 0.32 (P=0.009), that correlated to changes in PCr concentration (P<0.001) and to the PCr/beta-ATP ratio (P=0.01). The uptake was mainly (90-95%) located to 1-2 microm large extra cellular 'granule'. A picture resembling early necrosis was seen on the H & E stainings and reflected in the Billingham rejection score above. CONCLUSIONS: After 6 h of RP the onset of acute rejection in the concordant hamster xeno hearts displayed features of early, possibly mitochondrial, necrosis, but not apoptosis, which correlated to changes in the PCr concentration and the PCr/beta-ATP ratio. The mechanism for the early rejection observed is unclear and might be caused by other factors in the sera apart from cellular components, antibodies and complement factors. Identification of the underlying mechanisms could enable us to design rational therapies that prevent activation of the recipient's innate immune response. PMID- 14551031 TI - Simultaneous blockade of co-stimulatory signals, CD28 and ICOS, induced a stable tolerance in rat heart transplantation. AB - An inducible co-stimulator (ICOS), a recently identified co-stimulatory receptor with a close structural homology of CD28 and CTLA4, is expressed on activated T cells. Anti-ICOS antibody was demonstrated to be effective on prolongation of graft survival after liver transplantation in rats. In this study, we investigated the potency of tolerance induction using the antibody combined with a recombinant adenovirus vector containing CTLA-4Ig cDNA (AdCTLA-4Ig) in rat heart transplantation model. Using a DA-to-Lewis rat heart transplantation model, an anti-rat ICOS antibody and AdCTLA-4Ig were simultaneously administered i.v. into recipients. The tissue specimens from the grafts were removed on various days after transplantation for histological evaluation. Donor-strain skin and heart grafts, and third-party heart allografts were challenged in the recipients with a long-term surviving graft. Splenocytes from the tolerance-induced recipients were used for adoptive transfer study. Anti-ICOS antibody alone did not prolong the survival of heart allograft. AdCTLA-4Ig monotherapy significantly prolonged the survival of heart allograft (Group 4). With a combination of Anti ICOS antibody and AdCTLA-4Ig, all recipients were resulted in a long-term allograft acceptance for more than 200 days (Group 8). When challenged donor strain skin grafts in the tolerant rats of Group 4, the skin was rejected, which also lead to a rejection of primary heart allografts. The recipients in Group 8 also rejected donor-strain skin grafts with no rejection of the primary heart grafts. These recipients accepted secondary heart grafts from donor-strain but not third-party. In Group 8 long-term survival recipients showed a high population of CD4+CD25+ regulatory T cell in peripheral blood, and in adoptive transfer study subtraction of these CD4+CD25+ T cells accelerate the rejection of heart graft in secondary irradiated recipients. The present results demonstrated that anti-ICOS antibody combined with AdCTLA-4Ig potently induces a stable immune tolerance after heart allografting in rat, which is mediated by the induction of CD4+CD25+ regulatory T cells. This strategy may be attractive for clinical employment to induce transplantation tolerance. PMID- 14551032 TI - CD30 antigen: not a physiological marker for TH2 cells but an important costimulator molecule in the regulation of the balance between TH1/TH2 response. AB - Understanding the physiological role of CD30 would be an important step forward in transplants because CD30+ T cells can be induced by alloantigens even in the presence of immunosuppressives such as cyclosporine (Csa) and hence can act as regulatory cells in allograft. The results of functional studies on purified T CD30+ cell populations led us to hypothesize that the CD30 costimulator molecule is not a specific marker for TH2 cells in normal conditions, as has been suggested, but rather a marker for an important immunoregulatory subpopulation that regulates the balance between TH1 and TH2 (TH1/TH2) type response. To substantiate this hypothesis we studied the TH1/TH2 cytokine network in peripheral whole blood cultures stimulate with M44 CD30 ligand (CD30L), an agonistic monoclonal antibody (mAb). Four types of whole blood culture were used: the first had been stimulated with anti-CD3 mAb which generates a CD30 cytokine profile similar to alloreactive stimulation; the second with anti-CD3 mAb+M81 (an anti-CD30L mAb) to inhibit CD30/CD30L interaction; the third with anti-CD3+anti interleukin (IL)4 mAbs to counteract IL4 activity and the fourth with anti CD3+anti-interferon (IFN)gamma mAbs to counteract IFNgamma activity. Network interactions between soluble CD30 (sCD30, a maker of CD30 expression), sBcl2 (a marker of cell survival) and TH1/TH2 cytokines (IFNgamma, IL2, IL12p70, IL12p40, IL4, IL5 and IL10) were then studied in the supernatants obtained. Our results confirm the hypothesis above by showing that CD30 signals trigger functional mechanisms responsible for changes in levels of production of several important TH1 and TH2 cytokines involved in the regulation of the physiological balance between TH1/TH2 functions. The CD30-stimulated network, in fact, induces IFNgamma production linked to TH1 activity (-->TH1) which is subsequently integrated by IL4 production linked to TH2 activity (-->TH2). This production appears to be regulated, respectively, by IL12p40 (-->TH2) and IL12p70 (-->TH1) production which could maintain the balance between TH1/TH2 type response (TH1<-->TH2). Further CD30 mechanisms are the regulation of the interactions between: IL5 IFNgamma, IL5-IL4, IL2-IL10, IL2-IL12p40 and IL10-IL12p70 production. The immunoregulatory activity of CD30 was confirmed by the lack of production balance between the above-mentioned cytokines observed in cultures in which the interaction between CD30 and its natural ligand (CD30/CD30L) and IL4 or IFNgamma activity had been blocked. We therefore conclude that CD30 may be an important costimulatory molecule and marker for the physiological balance between TH1/TH2 immune response. Consequently, further study of CD30 immunoregulatory mechanisms may allow for the identification of methods for re-establishing equilibrium and hence more effective strategies for the prevention and treatment of immunopathological conditions such as transplant rejection. PMID- 14551033 TI - Inducible nitric oxide synthase inhibitors prolonged the survival of skin xenografts through selective down-regulation of pro-inflammatory cytokine and CC chemokine expressions. AB - To elucidate the possible immunoregulatory role of nitric oxide (NO) in cellular xenograft rejection we performed rat-to-mouse skin xenotransplantation. The rat skin engrafted mice were treated with the inducible NO synthase (iNOS) inhibitors, aminoguanidine (AMG, 200 mg/kg) and NG-nitro-L-arginine methyl ester (L-NAME, 60 mg/kg) every other day until rejection. Skin xenograft survival was monitored and immune cell infiltration and intragraft cytokine and chemokine mRNA expressions were analyzed 7 days after grafting. Compared with the control mice, the AMG- and L-NAME treated mice showed delayed xenograft rejection by approximately 3 days (8.9 +/- 0.7 days vs. 11.7 +/- 1.2 and 12.0 +/- 0.9 days, respectively). Infiltrations of CD11b+, MOMA-2+ cells and neutrophils were significantly reduced in both AMG- and L-NAME treated graft but CD4+ and CD8+ cells were not. The expression of cytokines such as IL-1beta, IL-2, IL-6, IL-12 and IFN-gamma in AMG- and L-NAME treated grafts were significantly decreased (P<0.01), whereas IL-10, TNF-alpha and TGF-beta1 were unchanged or enhanced. Additionally, the expressions of CC-chemokines, such as RANTES and MIP-1alpha, were significantly reduced (P<0.01) whereas the expressions of CXC-chemokines, such as IP-10 and MIG, were unchanged. These results imply that prolonged rat-to mouse skin xenograft survival by iNOS inhibitors may be due to the selective inhibition of pro-inflammatory cytokines and chemokines and suggest the possible regulatory role of NO in cytokine and chemokine expressions during xenotransplant rejection. PMID- 14551034 TI - HLA-A, B, C, DRB1, DQB1 matching heterogeneity in 'favourably matched' kidney recipients. AB - Allocation of cadaveric donor kidneys in the UK is founded on matching for HLA-A, -B and -DR, primarily at the broad specificity level. Increasing evidence shows that matching at a higher resolution and consideration of additional loci, such as HLA-C, -DQ and -DP, improves graft outcome. The aim of this study was to clarify the typical level of split specificity HLA-A, -B, -C, -DR, -DQ and allelic -DRB1 and -DQB1 mismatching in 'favourably matched' cadaveric renal transplant pairs. Two hundred and thirty-seven cadaveric donor/recipient pairs, 'favourably matched', according to United Kingdom Transplant criteria, were typed at the split specificity level for HLA-A, -B, -C and at the allele level for HLA DRB1 and -DQB1. The level of split specificity and allele mismatching was then assessed. Overall, 66.7% of the patients had at least one HLA-C mismatch with their donors; 36.9% of those matched for HLA-B and 85.5% of those mismatched for HLA-B (P<0.0001). A broad specificity HLA-A or -B mismatch influenced the presence of an HLA-B, or HLA-A split specificity mismatch, respectively, (P<0.05) but made no significant difference to the presence of an HLA-DR split mismatch. Overall, 4.6% of the patients were mismatched for HLA-DR split specificities but 30.4% were mismatched at HLA-DQ and 50.6% had at least one HLA-DRB1 or -DQB1 allele mismatch. Considerable HLA-A, -B, -C, -DR, -DQ matching heterogeneity exists even amongst 'well matched' renal transplant patient groups. Little is known about the effects of combinations of mismatched specificities on graft survival. Thus, further investigation is merited particularly for HLA-C and -DQ mismatching. PMID- 14551036 TI - House keeping genes and gene expression analysis in transplant recipients: a note of caution. AB - BACKGROUND: House keeping genes are often used as a means of standardising results obtained in gene expression investigations. This study was performed to investigate whether beta-actin, beta2-microglobulin (two genes frequently quoted as house keeping genes) and/or transferrin receptor would be suitable house keeping genes for use in gene expression analysis of renal transplant recipients. METHODS: Sequential expression of all three genes was measured in the peripheral blood mononuclear cells of 13 living donors and 45 renal transplant recipients, pre-operatively and then daily for up to 2 weeks. Fifteen of the recipients experienced an episode of biopsy proven acute rejection. Gene expression measurement was performed using quantitative real time 'TaqMan' PCR technology. RESULTS: Gene expression of all three genes was unchanged in the living donor cohort. However, in the transplant recipients there were significant increases in expression following transplantation in the non-rejectors, and preceding the diagnosis of acute rejection. In the latter group, levels returned to pre transplant values after the commencement of anti-rejection therapy. CONCLUSIONS: Beta-actin, beta2-microglobulin and transferrin receptor gene expression, although not influenced by surgery, is influenced by transplantation, acute rejection and anti-rejection therapy making these genes unsuitable as house keeping genes following renal transplantation. These findings may cast doubt on the results of some studies that used these genes for the purposes of standardisation when looking at cDNA measurement. We suggest that any group wishing to use a house keeping gene ensure that its expression is independent of study parameters prior to the start of the study. PMID- 14551035 TI - Positive and negative selection of T cell repertoires during differentiation in allogeneic bone marrow chimeras. AB - T cells acquire immune functions during expansion and differentiation in the thymus. Mature T cells respond to peptide antigens (Ag) derived from foreign proteins when these peptide Ag are presented on the self major histocompatibility complex (MHC) molecules but not on allo-MHC. This is termed self-MHC restriction. On the other hand, T cells do not induce aggressive responses to self Ag (self tolerance). Self-MHC restriction and self-tolerance are not genetically determined but acquired a posteriori by positive and negative selection in the thymus in harmony with the functional maturation. Allogeneic bone marrow (BM) chimera systems have been a useful strategy to elucidate mechanisms underlying positive and negative selection. In this communication, the contribution of BM chimera systems to the investigation of the world of T-ology is discussed. PMID- 14551037 TI - Short-term exposure of Chinook salmon (Oncoryhnchus tshawytscha) to o,p-DDE or DMSO during early life-history stages causes long-term humoral immunosuppression. AB - We evaluated the effect of short-term exposures to a xenobiotic chemical during early life-history stages on the long-term immune competence of chinook salmon (Oncoryhnchus tshawytscha). Immersion of chinook salmon eggs in a nominal concentration of o,p-dichlorodiphenyldichloroethylene (o,p-DDE; 10 ppm) for 1 hr at fertilization followed by immersion in the same dose for 2 hr at hatch resulted in a significant reduction in the ability of splenic leukocytes from fish 1 year after treatment to undergo blastogenesis upon in vitro stimulation with lipopolysaccharide. We also observed that the vehicle, dimethyl sulfoxide (DMSO), caused a significant reduction in the ability of the splenic leukocytes to express surface immunoglobin M (SIgM) at this time. The concentration of o,p DDE in a pooled sample of whole fry from this treatment was 0.53 microg/g lipid 1 month after first feeding but was undetectable in all other treatments. Mortality rate, time to hatch, fish length, and weight were unaffected by treatment with o,p-DDE. Similarly, sex ratios, gonadal development, and concentrations of plasma estradiol and 11-ketotestosterone were not affected by the treatment. In addition, we found no evidence that plasma lysozyme concentrations or the mitogenic responses of splenic leukocytes to concanavalin A or polyinosinic polycytidylic acid were influenced by the treatment. In this experiment, a brief period of exposure to o,p-DDE or DMSO during early development was able to induce long-term effects on humoral immune competence of chinook salmon. Such immunosuppression may increase susceptibility to disease, which may in turn be critical to regulating the population. PMID- 14551038 TI - Guanylyl cyclases, nitric oxide, natriuretic peptides, and airway smooth muscle function. AB - Airway smooth muscle (ASM) plays an important role in asthma pathophysiology through its contractile and proliferative functions. The cyclic nucleotides adenosine 3',5'-cyclic monophosphate (cAMP) and guanosine 3',5'-cyclic monophosphate (cGMP) are second messengers capable of mediating the effects of a variety of drugs and hormones. There is a large body of evidence to support the hypothesis that cAMP is a mediator of the ASM's relaxant effects of drugs, such as beta2-adrenoceptor agonists, in human airways. Although most attention has been paid to this second messenger and the signal transduction pathways it activates, recent evidence suggests that cGMP is also an important second messenger in ASM with important relaxant and antiproliferative effects. Here, we review the regulation and function of cGMP in ASM and discuss the implications for asthma pathophysiology and therapeutics. Recent studies suggest that activators of soluble and particulate guanylyl cyclases, such as nitric oxide donors and natriuretic peptides, have both relaxant and antiproliferative effects that are mediated through cGMP-dependent and cGMP-independent pathways. Abnormalities in these pathways may contribute to asthma pathophysiology, and therapeutic manipulation may complement the effects of beta2-adrenoceptor agonists. PMID- 14551039 TI - New role for glycocalyx in lung. PMID- 14551040 TI - MRI and echocardiographic assessment of the diastolic dysfunction of normal aging: altered LV pressure decline or load? AB - Changes in diastolic indexes during normal aging, including reduced early filling velocity (E), lengthened E deceleration time (DT), augmented late filling (A), and prolonged isovolumic relaxation time (IVRT), have been attributed to slower left ventricular (LV) pressure (LVP) decay. Indeed, this constellation of findings is often referred to as the "abnormal relaxation" pattern. However, LV filling is determined by the atrioventricular pressure gradient, which depends on both LVP decline and left atrial (LA) pressure (LAP). To assess the relative influence of LVP decline and LAP, we studied 122 normal subjects aged 21-92 yr by Doppler echocardiography and MRI. LVP decline was assessed by color M-mode (V(p)) and the LV untwisting rate. Early diastolic LAP was evaluated using pulmonary vein flow systolic fraction, pulmonary vein flow diastolic DT, color M-mode (E/V(p)), and tissue Doppler (E/E(m)). Linear regression showed the expected reduction of E, increase in A, and prolongation of IVRT and DT with advancing age. There was no relation of age to parameters reflecting the rate of LVP decline. However, older age was associated with reduced E/V(p) (P = 0.008) and increased pulmonary vein systolic fraction (P < 0.001), pulmonary vein DT (P = 0.0026), and E/E(m) (P < 0.0001), all suggesting reduced early LAP. Therefore, reduced early filling in older adults may be more closely related to a reduced early diastolic LAP than to slower LVP decline. This effect also explains the prolonged IVRT. We postulate that changes in LA active or passive properties may contribute to development of the abnormal relaxation pattern during the aging process. PMID- 14551041 TI - Evidence of a role for TRPC channels in VEGF-mediated increased vascular permeability in vivo. AB - Vascular endothelial growth factor (VEGF) increases vascular permeability by stimulating endothelial Ca(2+) influx. Here we provide evidence that links VEGF mediated increased permeability and endothelial intracellular Ca(2+) concentration ([Ca(2+)](i)) with diacylglycerol (DAG)-mediated activation of the transient receptor potential channels (TRPCs). We used the Landis-Michel technique to measure changes in hydraulic conductivity (L(p)) and fluorescence photometry to quantify changes in endothelial [Ca(2+)](i) in individually perfused Rana mesenteric microvessels in vivo and transfected nonendothelial cells in vitro. The membrane-permeant DAG analog 1-oleoyl-2-acetyl-sn-glycerol (OAG, 100 microM), which is known to increase Ca(2+) influx through TRPCs, transiently increased L(p) 3.8 +/- 1.2-fold (from 1.6 +/- 0.8 to 9.8 +/- 2.7 x 10(-7) cm.s(-1).cmH(2)O(-1); P < 0.0001; n = 18). Protein kinase C inhibition by bisindolylmaleimide (1 microM) did not affect the OAG-induced increases in L(p). OAG also significantly increased microvascular endothelial [Ca(2+)](i) in vivo (n = 13; P < 0.0001), which again was not sensitive to protein kinase C inhibition. VEGF induced a transient increase in endothelial [Ca(2+)](i) in human embryonic kidney cells (HEK-293) that were cotransfected with VEGF receptor 2 and TRPC-6 but not with control, VEGF receptor 2, or TRPC-6 expression vector alone (P < 0.01; n = 9). Flufenamic acid, which has been shown to enhance activity of TRPC-6 but inhibit TRPC-3 and -7, enhanced the VEGF-mediated increase in L(p) in approximately half of the vessels tested but inhibited the response in the other half of the vessels. These data provide evidence consistent with the hypothesis that VEGF increases vascular permeability via DAG-mediated Ca(2+) entry through TRPCs. Although the exact identities of the TRPCs remain to be confirmed, TRPC-6 appears to be a likely candidate in approximately half of the vessels. PMID- 14551042 TI - Cortisol-mediated regulation of uterine artery contractility: effect of chronic hypoxia. AB - We previously demonstrated that cortisol regulated alpha(1)-adrenoceptor-mediated contractions differentially in nonpregnant and pregnant uterine arteries. Given that chronic hypoxia during pregnancy has profound effects on maternal uterine artery reactivity, the present study investigated the effects of chronic hypoxia on cortisol-mediated regulation of uterine artery contractions. Pregnant (day 30) and nonpregnant ewes were divided between normoxic control and chronically hypoxic [maintained at high altitude (3,820 m), arterial Po(2): 60 mmHg for 110 days] groups. Uterine arteries were isolated and contractions measured. In hypoxic animals, cortisol (10 ng/ml for 24 h) increased norepinephrine-induced contractions in pregnant, but not in nonpregnant, uterine arteries. The 11beta hydroxysteroid dehydrogenase inhibitor carbenoxolone did not change cortisol effects in nonpregnant uterine arteries, but abolished it in pregnant uterine arteries by increasing norepinephrine pD(2) (-log EC(50)) in control tissues. The dissociation constant of norepinephrine-alpha(1)-adrenoceptors was not changed by cortisol in nonpregnant, but decreased in pregnant uterine arteries. There were no differences in the density of glucocorticoid receptors between normoxic and hypoxic tissues. Cortisol inhibited the norepinephrine-induced increase in Ca(2+) concentrations in nonpregnant arteries, but potentiated it in pregnant arteries. In addition, cortisol attenuated phorbol 12,13-dibutyrate-induced contractions in normoxic nonpregnant and pregnant uterine arteries, but had no effect on the contractions in hypoxic arteries. The results suggest that cortisol differentially regulates alpha(1)-adrenoceptor- and PKC-mediated contractions in uterine arteries. Chronic hypoxia suppresses uterine artery sensitivity to cortisol, which may play an important role in the adaptation of uterine vascular tone and blood flow in response to chronic stress of hypoxia during pregnancy. PMID- 14551043 TI - The intermediary metabolite pyruvate attenuates stunning and reduces infarct size in in vivo porcine myocardium. AB - The intermediary metabolite pyruvate has been shown to exert significant beneficial effects in in vitro models of myocardial oxidative stress and ischemia reperfusion injury. However, there have been few reports of the ability of pyruvate to attenuate myocardial stunning or reduce infarct size in vivo. This study tested whether supraphysiological levels of pyruvate protect against reversible and irreversible in vivo myocardial ischemia-reperfusion injury. Anesthetized, open-chest pigs (n = 7/group) underwent 15 min of left anterior descending coronary artery (LAD) occlusion and 3 h of reperfusion to induce stunning. Load-insensitive contractility measurements of regional preload recruitable stroke work (PRSW) and PRSW area (PRSWA) were generated. Vehicle or pyruvate (100 mg/kg i.v. bolus + 10 mg x kg(-1) x min(-1) intra-atrial infusion) was administered during ischemia and for the first hour of reperfusion. In infarct studies, pigs (n = 6/group) underwent 1 h of LAD ischemia and 3 h of reperfusion. Group I pigs received vehicle or pyruvate for 30 min before and throughout ischemia. In group II, the infusion was extended through 1 h of reperfusion. In the stunning protocol, pyruvate significantly improved the recovery of PRSWA at 1 h (50 +/- 4% vs. 23 +/- 3% in controls) and 3 h (69 +/- 5% vs. 39 +/- 3% in controls) reperfusion. Control pigs exhibited infarct sizes of 66 +/- 1% of the area at risk. The pyruvate I protocol was associated with an infarct size of 49 +/- 3% (P < 0.05), whereas the pyruvate II protocol was associated with an infarct size of 30 +/- 2% (P < 0.05 vs. control and pyruvate I). These findings suggest that pyruvate attenuates stunning and decreases myocardial infarction in vivo in part by reduction of reperfusion injury. Metabolic interventions such as pyruvate should be considered when designing the optimal therapeutic strategies for limiting myocardial ischemia-reperfusion injury. PMID- 14551044 TI - Heat shock protein 90 and tyrosine kinase regulate eNOS NO* generation but not NO* bioactivity. AB - An increase in the association of heat shock protein 90 (HSP90) with endothelial nitric oxide (NO) synthase (eNOS) is well recognized for increasing NO (NO*) production. Despite the progress in this field, the mechanisms by which HSP90 modulates eNOS remain unclear due, in part, to the fact that geldanamycin (GA) redox cycles to generate superoxide anion (O(2)(-*) and the fact that inhibiting HSP90 with GA or radicicol (RAD) destabilizes tyrosine kinases that rely on the chaperone for maturation. In this report, we determine the extent to which these side effects alter vascular and endothelial cell function in physiologically relevant systems and in cultured endothelial cells. Vascular endothelial growth factor (VEGF)-stimulated vascular permeability, as measured by Evans blue leakage in the ears of male Swiss mice in vivo, and acetylcholine-induced vasodilation of isolated, pressurized mandibular arterioles from male C57BL6 mice ex vivo were attenuated by N(omega)-nitro-L-arginine methyl ester (L-NAME), GA, and RAD. Z-1[N (2-aminoethyl)-N-(2-ammonoethyl)amino]diazen-1-ium-1,2-dioate (DETA-NONOate), a slow releasing NO. donor, increased vasodilation of arterioles pretreated with GA, RAD, and L-NAME equally well except at 10(-5) M, the highest concentration used, where vasodilation was greater in pressurized arterioles treated with L NAME than in arterioles pretreated with GA or RAD alone. Both GA and RAD reduced NO* release from stimulated endothelial cell cultures and increased O(2)(-*) production in the endothelium of isolated aortas by an L-NAME-inhibitable mechanism. Pretreatment with RAD increased stimulated O(2)(-*) production from eNOS, whereas pretreatment with genistein (GE), a broad-spectrum tyrosine kinase inhibitor, did not; however, pretreatment with GE + RAD resulted in a super induced state of uncoupled eNOS activity upon stimulation. These data suggest that the tyrosine kinases, either directly or indirectly, and HSP90-dependent signaling pathways act in concert to suppress uncoupled eNOS activity. PMID- 14551045 TI - PPARgamma activation, by reducing oxidative stress, increases NO bioavailability in coronary arterioles of mice with Type 2 diabetes. AB - We tested the hypothesis that short-term treatment of mice with Type 2 diabetes mellitus (DM) with rosiglitazone (ROSI), an agonist of peroxisome proliferator activated receptor-gamma, ameliorates the impaired coronary arteriolar dilation by reducing oxidative stress via a mechanism unrelated to its effect on hyperglycemia and hyperinsulinemia. Control and Type 2 DM (db/db) mice were treated with ROSI (3 mg x kg(-1) x day(-1)) for 7 days, which did not significantly affect their serum concentration of glucose and insulin. Compared with controls, in db/db mice serum levels of 8-isoprostane and dihydroethydine detectable superoxide production in carotid arteries were significantly elevated and were reduced by ROSI treatment. In coronary arterioles (diameter, approximately 80 microm) isolated from db/db mice, the reduced dilations to ACh, the nitric oxide (NO) donor NONOate, and increases in flow were significantly augmented either by in vitro administration of apocynin, an inhibitor of NAD(P)H oxidase, or by in vivo ROSI treatment, responses that were then significantly reduced by the NO synthase inhibitor N(omega)-nitro-L-arginine methyl ester. In aortas of db/db mice, activity of SOD and catalase was reduced, whereas NAD(P)H oxidase activity was enhanced. ROSI treatment enhanced catalase and reduced NAD(P)H oxidase activity but did not affect the activity of SOD. These findings suggest that ROSI treatment enhances NO mediation of coronary arteriolar dilations due to the reduction of vascular NAD(P)H oxidase-derived superoxide production and enhancement of catalase activity. Thus, in addition to the previously revealed beneficial metabolic effects, the antioxidant action of rosiglitazone may protect coronary arteriolar function in Type 2 DM. PMID- 14551047 TI - Testosterone suppresses endothelium-dependent dilation of rat middle cerebral arteries. AB - Little is known about vascular effects of testosterone. We previously reported chronic testosterone treatment increases vascular tone in middle cerebral arteries (MCA; 300 microm diameter) of male rats. In the present study, we investigated the hypothesis that physiological levels of circulating testosterone affect endothelial factors that modulate cerebrovascular reactivity. Small branches of MCA (150 microm diameter) were isolated from orchiectomized (ORX) and testosterone-treated (ORX+T) rats. Intraluminal diameters were recorded after step changes in intraluminal pressure (20-100 Torr) in the absence or presence of N(G)-nitro-L-arginine-methyl ester (L-NAME), a nitric oxide synthase (NOS) inhibitor; indomethacin, a cyclooxygenase (COX) inhibitor; and/or apamin and charybdotoxin (CTX); and K(Ca) channel blockers used to inhibit endothelium derived hyperpolarizing factors (EDHF). At intraluminal pressures >or=60 Torr, arteries from ORX+T developed greater tone compared with ORX arteries. This difference was abolished by removal of the endothelium but remained after treatment of intact arteries with indomethacin or L-NAME. In addition, testosterone treatment had no effect on cerebrovascular production of endothelin 1 or prostacyclin nor did it alter protein levels of endothelial NOS or COX-1. Endothelium removal after L-NAME/indomethacin exposure caused an additional increase in tone. Interestingly, the latter effect was smaller in arteries from ORX+T, suggesting testosterone affects endothelial vasodilators that are independent of NOS and COX. Apamin/CTX, in the presence of L-NAME/indomethacin, abolished the difference in tone between ORX and ORX+T and resulted in vessel diameters similar to those of endothelium-denuded preparations. In conclusion, testosterone may modulate vascular tone in cerebral arteries by suppressing EDHF. PMID- 14551046 TI - In vivo expression and function of recombinant GTPCH I in the rabbit carotid artery. AB - Tetrahydrobiopterin (BH4) is an essential co-factor for endothelial nitric oxide synthase enzymatic activity. GTP cyclohydrolase I (GTPCH I) is the rate-limiting enzyme in BH4 synthesis. This study set out to test the hypothesis that in vivo gene transfer of GTPCH I to endothelial cells could increase bioavailability of BH4, enhance biosynthesis of nitric oxide and thereby enhance endothelium dependent relaxations mediated by nitric oxide. In vivo gene transfer was carried out by adenovirus (Ad)-mediated delivery into rabbit carotid arteries. Each artery was transduced by 20-min intraluminal incubation of 10(9) plaque-forming units of Ad-encoding GTPCH I (AdGTPCH) or beta-galactosidase as a control. The rabbits were euthanized 72 h later, and vasomotor function of isolated arteries was assessed by isometric force recording. GTPCH I enzymatic activity, BH4, and oxidized biopterin levels were detected with the use of HPLC, and cGMP was measured with the use of radioimmunoassay. Expression of recombinant proteins was detected predominantly in endothelial cells. Both GTPCH I activity and BH4 levels were increased in arteries transduced with AdGTPCH. However, contraction to phenylephrine (10(-5) to 10(-9) M), endothelium-dependent relaxation to acetylcholine (10(-5) to 10(-9) M) and cGMP levels were not significantly affected by increased expression of GTPCH I. Our results suggest that expression of GTPCH I in vascular endothelium in vivo increases intracellular concentration of BH4. However, under physiological conditions, it appears that this increase does not affect nitric oxide production in endothelial cells of the carotid artery. PMID- 14551048 TI - Inhibition of glucose uptake in murine cardiomyocyte cell line HL-1 by cardioprotective drugs dilazep and dipyridamole. AB - Inhibition of adenosine reuptake by nucleoside transport inhibitors, such as dipyridamole and dilazep, is proposed to increase extracellular levels of adenosine and thereby potentiate adenosine receptor-dependent pathways that promote cardiovascular health. Thus adenosine can act as a paracrine and/or autocrine hormone, which has been shown to regulate glucose uptake in some cell types. However, the role of adenosine in modulating glucose transport in cardiomyocytes is not clear. Therefore, we investigated whether exogenously applied adenosine or inhibition of adenosine transport by S-(4-nitrobenzyl)-6 thioinosine (NBTI), dipyridamole, or dilazep modulated basal and insulin stimulated glucose uptake in the murine cardiomyocyte cell line HL-1. HL-1 cell lysates were subjected to SDS-PAGE and immunoblotting to determine which GLUT isoforms are present. Glucose uptake was measured in the presence of dipyridamole (3-300 microM), dilazep (1-100 microM), NBTI (10-500 nM), and adenosine (50-250 microM) or the nonmetabolizable adenosine analog 2-chloro-adenosine (250 microM). Our results demonstrated that HL-1 cells possess GLUT1 and GLUT4, the isoforms typically present in cardiomyocytes. We found no evidence for adenosine-dependent regulation of basal or insulin-stimulated glucose transport in HL-1 cardiomyocytes. However, we did observe a dose-dependent inhibition of glucose transport by dipyridamole (basal, IC(50) = 12.2 microM, insulin stimulated, IC(50) = 13.09 microM) and dilazep (basal, IC(50) = 5.7 microM, insulin stimulated, IC(50) = 19 microM) but not NBTI. Thus our data suggest that dipyridamole and dilazep, which are widely used to specifically inhibit nucleoside transport, have a broader spectrum of transport inhibition than previously described. Moreover, these data may explain previous observations, in which dipyridamole was noted to be proischemic at high doses. PMID- 14551050 TI - Cedrelopsis grevei induced hypotension and improved endothelial vasodilatation through an increase of Cu/Zn SOD protein expression. AB - This study was designed to investigate the cardiovascular consequences of oral administration of Cedrelopsis grevei (CG) in normotensive rats. Experiments were designed to investigate hemodynamic parameters in vivo as well as the consequences of CG treatment on the vasoconstriction response to norepinephrine and the vasorelaxant response to ACh ex vivo in isolated aortas and small mesenteric arteries (SMA). Treatment of male Wistar rats with 80 mg/kg CG for 4 wk induced a progressive decrease in systolic blood pressure. In the aorta, CG did not significantly alter the response to norepinephrine despite the participation of extraendothelial nitric oxide (NO)-induced hyporeactivity. In the SMA, contraction to norepinephrine was not modified by CG treatment even though it enhanced the participation of endothelial NO. Endothelium-dependent relaxation to ACh was increased in both the aorta and SMA from CG-treated rats. In the aorta from CG-treated rats, the mechanism involved superoxide dismutase (SOD)- and catalase-sensitive free radical production. The latter was associated with enhanced expression of Cu/Zn SOD and endothelial NO synthase. These results suggest that oral administration of CG produces a decrease in blood pressure in normotensive rats. This hemodynamic effect was associated with enhanced endothelium-dependent relaxation and an induction of Cu/Zn SOD and endothelial NO synthase expressions in the vessel wall. They also show subtle mechanisms that compensate for the increased participation of NO to maintain unchanged agonist induced contractility. These data provide a pharmacological basis for the empirical use of CG against cardiovascular diseases. PMID- 14551049 TI - Redox modulation of the inotropic response to dobutamine is impaired in patients with heart failure. AB - It has been suggested that oxidative stress contributes to impaired left ventricular (LV) contractility in the setting of heart failure (HF). To test this hypothesis, we studied the effect of an antioxidant on contractility at rest and in response to dobutamine in 10 HF patients. We hypothesized that vitamin C would augment contractility in HF and that this effect would be of a greater magnitude in HF patients compared with patients with normal LV (NLV) function. Data from 10 patients with NLV function who participated in this study are included in this report and have been published elsewhere. A micromanometer-tipped catheter was introduced into the LV. In the experimental protocol, an infusion catheter was positioned in the left main coronary artery. The peak positive rate of change of LV pressure (LV +dP/dt) was measured in response to the intravenous infusion of dobutamine before and during the intracoronary infusion of vitamin C (96 mg/min). Vitamin C had no effect on basal LV +dP/dt in either HF or NLV groups. The infusion of vitamin C augmented the LV +dP/dt response to dobutamine by 22 +/- 4% in the NLV function group. In contrast, vitamin C had no effect on the inotropic response to dobutamine in the HF group. In the control protocol, without vitamin C, no differences were observed between responses to two sequential dobutamine infusions in either group (HF, n = 11; NLV, n = 9). Therefore, a positive effect of vitamin C on contractility was limited to patients with NLV function. The absence of this effect in HF patients may suggest that normal redox responsiveness is lost in this disease state. PMID- 14551051 TI - Strain softening is not present during axial extensions of rat intact right ventricular trabeculae in the presence or absence of 2,3-butanedione monoxime. AB - Recent studies of passive myocardial mechanics have shown that strain softening behavior is present during both inflation of isolated whole rat hearts and shearing of tissue blocks taken from the left ventricular free wall in pigs. Strain softening is typically manifested by a stiffer force-extension relation in the first deformation cycle relative to subsequent cycles and is distinguished from viscoelasticity by a lack of recovery of stiffness, even after several hours of rest. The causes of this behaviour are unknown. We investigated whether strain softening is observed in uniaxial extensions of intact, viable, rat right ventricular (RV) cardiac trabeculae. Stretch and release cycles of 5%, 10%, and 15% muscle length were applied at a constant velocity at 26 degrees C. Muscles were tested in random order in the presence and absence of 50 mM 2,3-butanedione monoxime (BDM). Whereas strain softening was displayed by nonviable trabeculae, it was not observed in viable preparations undergoing physiologically relevant extensions whether in the presence or absence of BDM. BDM also had no effect on passive compliance. There was a reversible increase of muscle compliance between the first and subsequent cycles, with recovery after 30 s of rest, independent of the presence of BDM. We conclude that strain softening is neither intrinsic to viable rat RV trabeculae nor influenced by BDM and that passive trabeculae compliance is not altered by the addition of BDM. PMID- 14551052 TI - Transmural left ventricular mechanics underlying torsional recoil during relaxation. AB - Early relaxation in the cardiac cycle is characterized by rapid torsional recoil of the left ventricular (LV) wall. To elucidate the contribution of the transmural arrangement of the myofiber to relaxation, we determined the time course of three-dimensional fiber-sheet strains in the anterior wall of five adult mongrel dogs in vivo during early relaxation with biplane cineangiography (125 Hz) of implanted transmural markers. Fiber-sheet strains were found from transmural fiber and sheet orientations directly measured in the heart tissue. The strain time course was determined during early relaxation in the epicardial, midwall, and endocardial layers referenced to the end-diastolic configuration. During early relaxation, significant circumferential stretch, wall thinning, and in-plane and transverse shear were observed (P < 0.05). We also observed significant stretch along myofibers in the epicardial layers and sheet shortening and shear in the endocardial layers (P < 0.01). Importantly, predominant epicardial stretch along the fiber direction and endocardial sheet shortening occurred during isovolumic relaxation (P < 0.05). We conclude that the LV mechanics during early relaxation involves substantial deformation of fiber and sheet structures with significant transmural heterogeneity. Predominant epicardial stretch along myofibers during isovolumic relaxation appears to drive global torsional recoil to aid early diastolic filling. PMID- 14551053 TI - Do P2X purinergic receptors regulate skeletal muscle blood flow during exercise? AB - Although there is evidence that sympathetic nerves release ATP as a neurotransmitter to produce vasoconstriction via P2X purinergic receptors, the role of these receptors in the regulation of blood flow to exercising skeletal muscle has yet to be determined. We hypothesized that there is tonic P2X receptor mediated vasoconstriction in exercising skeletal muscle. To test this hypothesis, the effect of P2X receptor blockade on skeletal muscle blood flow was examined in six exercising mongrel dogs. P2X receptor antagonism was accomplished with pyridoxal-phosphate-6-azophenyl-2'4'-disulfonic acid (PPADs). Animals were instrumented chronically with flow probes on the external iliac arteries of both hindlimbs and a catheter in one femoral artery. PPADs (40 mg) was infused as a bolus into the femoral artery catheter during steady-state exercise at 6 miles/h. Intra-arterial infusion of PPADs increased iliac blood flow from 542 +/- 55 to 677 +/- 69 ml/min (P < 0.05) and iliac vascular conductance from 5.17 +/- 0.62 to 6.53 +/- 0.80 ml.min(-1).mmHg(-1). The PPADs infusion did not affect blood flow in the contralateral iliac artery. These data support the hypothesis that P2X purinergic receptors produce vasoconstriction in exercising skeletal muscle. PMID- 14551054 TI - Quantification of left ventricular mechanical dyssynchrony by conductance catheter in heart failure patients. AB - Mechanical dyssynchrony is an important codeterminant of cardiac dysfunction in heart failure. Treatment, either medical, surgical, or by pacing, may improve cardiac function partly by improving mechanical synchrony. Consequently, the quantification of ventricular mechanical (dys)synchrony may have important diagnostic and prognostic value and may help to determine optimal therapy. Therefore, we introduced new indexes to quantify temporal and spatial aspects of mechanical dyssynchrony derived from online segmental conductance catheter signals obtained during diagnostic cardiac catheterization. To test the feasibility and usefulness of our approach, we determined cardiac function and left ventricular mechanical dyssynchrony by the conductance catheter in heart failure patients with intraventricular conduction delay (n = 12) and in patients with coronary artery disease (n = 6) and relatively preserved left ventricular function. The heart failure patients showed depressed systolic and diastolic function. However, the most marked hemodynamic differences between the groups were found for mechanical dyssynchrony, indicating a high sensitivity and specificity of the new indexes. Comparison of conductance catheter-derived indexes with septal-to-lateral dyssynchrony derived by tissue-Doppler velocity imaging showed highly significant correlations. The proposed indexes provide additional, new, and quantitative information on temporal and spatial aspects of mechanical dyssynchrony. They may refine diagnosis of cardiac dysfunction and evaluation of interventions, and ultimately help to select optimal therapy. PMID- 14551056 TI - Increased focal Kv4.2 channel expression at the plasma membrane is the result of actin depolymerization. AB - Voltage-dependent potassium channel trafficking and localization are regulated by proteins of the cytoskeleton, but the mechanisms by which these occur are still unclear. Using human embryonic kidney (HEK) cells as a heterologous expression system, we tested the role of the actin cytoskeleton in modulating the function of Kv4.2 channels. Pretreatment (>or=1 h) of HEK cells with 5 microM cytochalasin D to disrupt the actin microfilaments greatly augmented whole cell Kv4.2 currents at potentials positive to -20 mV. However, no changes in the voltage dependence of activation and inactivation of macroscopic currents were observed to account for this increase. Similarly, single channel recordings failed to reveal any significant changes in the single channel conductance, open probability, and kinetics. However, the mean patch current was increased from 0.9 +/- 0.2 pA in control to 6.7 +/- 3.0 pA in the presence of cytochalasin D. Imaging experiments revealed a clear increase in the surface expression of the channels and the appearance of "bright spot" features, suggesting that large numbers of channels were being grouped at specific sites. Our data provide clear evidence that increased numbers and altered distribution of Kv4.2 channels at the cell surface are primarily the result of reorganization of the actin cytoskeleton. PMID- 14551055 TI - Phosphatidylinositol 3-kinase modulates vascular smooth muscle contraction by calcium and myosin light chain phosphorylation-independent and -dependent pathways. AB - Regulation of smooth muscle contraction involves a number of signaling mechanisms that include both kinase and phosphatase reactions. The goal of the present study was to determine the role of one such kinase, phosphatidylinositol (PI)3-kinase, in vascular smooth muscle excitation-contraction coupling. Using intact medial strips of the swine carotid artery, we found that inhibition of PI3-kinase by LY 294002 resulted in a concentration-dependent decrease in the contractile response to both agonist stimulation and membrane depolarization-dependent contractions and a decrease in Ca(2+)-dependent myosin light chain (MLC) phosphorylation, the primary step in the initiation of smooth muscle contraction. Inhibition of PI3 kinase also depressed phorbol dibutyrate-induced contractions, which are not dependent on either Ca(2+) or MLC phosphorylation but are dependent on protein kinase C. To determine the Ca(2+)-dependent site of action of PI3-kinase, we determined the effect of several inhibitors of calcium metabolism on LY-294002 dependent inhibition of contraction. These inhibitors included nifedipine, SK&F 96365, and caffeine. Only SK&F-96365 blocked the LY-294002-dependent inhibition of contraction. Interestingly, all compounds blocked the LY-294002-dependent inhibition of MLC phosphorylation. Our results suggest that activation of PI3 kinase is involved in a Ca(2+)- and MLC phosphorylation-independent pathway for contraction likely to involve protein kinase C. In addition, our results also suggest that activation of PI3-kinase is involved in Ca(2+)-dependent signaling at the level of receptor-operated calcium channels. PMID- 14551057 TI - Intracellular calcium handling heterogeneities in intact guinea pig hearts. AB - Regional heterogeneities of ventricular repolarizing currents and their role in arrhythmogenesis have received much attention; however, relatively little is known regarding heterogeneities of intracellular calcium handling. Because repolarization properties and contractile function are heterogeneous from base to apex of the intact heart, we hypothesize that calcium handling is also heterogeneous from base to apex. To test this hypothesis, we developed a novel ratiometric optical mapping system capable of measuring calcium fluorescence of indo-1 at two separate wavelengths from 256 sites simultaneously. With the use of intact Langendorff-perfused guinea pig hearts, ratiometric calcium transients were recorded under normal conditions and during administration of known inotropic agents. Ratiometric calcium transients were insensitive to changes in excitation light intensity and fluorescence over time. Under control conditions, calcium transient amplitude near the apex was significantly larger (60%, P < 0.01) compared with the base. In contrast, calcium transient duration was significantly longer (7.5%, P < 0.03) near the base compared with the apex. During isoproterenol (0.05 microM) and verapamil (2.5 microM) administration, ratiometric calcium transients accurately reflected changes in contractile function, and, the direction of base-to-apex heterogeneities remained unchanged compared with control. Ratiometric optical mapping techniques can be used to accurately quantify heterogeneities of calcium handling in the intact heart. Significant heterogeneities of calcium release and sequestration exist from base to apex of the intact heart. These heterogeneities are consistent with base-to apex heterogeneities of contraction observed in the intact heart and may play a role in arrhythmogenesis under abnormal conditions. PMID- 14551058 TI - Shear stress and VEGF activate IKK via the Flk-1/Cbl/Akt signaling pathway. AB - Vascular endothelial cells are continuously exposed to mechanical (e.g., shear stress) and chemical (e.g., growth factors) stimuli. It is important to elucidate the mechanisms by which cells perceive and integrate these different stimuli to regulate the downstream signaling pathways. We (50) have previously reported the shear-induced interplay between two membrane receptors, integrins and Flk-1. In the present study, we investigated the molecular mechanisms regulating the downstream IkappaB kinase (IKK) pathway in response to shear stress and VEGF. Both shear stress and VEGF induced a transient increase of IKK activity. These effects were inhibited by SU-1498, a specific Flk-1 inhibitor, and by a negative mutant of Casitas B-lineage lymphoma (Cbl) with tyrosine-to-phenylalanine mutations at sites 700, 731, and 774 (Cbl(nm)). Because Flk-1 and Cbl form a complex upon shearing or VEGF applications (50), these results suggest that shear stress and VEGF activate IKK via the receptor Flk-1 and its recruitment of the adapter protein Cbl. The inhibition of the shear- and VEGF-induced IKK activities by a negative mutant of Akt indicates that Akt acts upstream to IKK in response to shear stress and VEGF. Furthermore, SU-1498 and Cbl(-nm) abolished the shear- and VEGF-induced Akt activity, indicating that Akt acts at a level downstream to Flk-1 and Cbl. Therefore, our results indicate that the signaling events induced by shear stress and VEGF converge at the membrane receptor Flk-1 and that these stimuli share the Flk-1/Cbl/Akt pathway in activating IKK activation. PMID- 14551059 TI - Medium perfusion enables engineering of compact and contractile cardiac tissue. AB - We hypothesized that functional constructs with physiological cell densities can be engineered in vitro by mimicking convective-diffusive oxygen transport normally present in vivo. To test this hypothesis, we designed an in vitro culture system that maintains efficient oxygen supply to the cells at all times during cell seeding and construct cultivation and characterized in detail construct metabolism, structure, and function. Neonatal rat cardiomyocytes suspended in Matrigel were cultured on collagen sponges at a high initial density (1.35 x 10(8) cells/cm(3)) for 7 days with interstitial flow of medium; constructs cultured in orbitally mixed dishes, neonatal rat ventricles, and freshly isolated cardiomyocytes served as controls. Constructs were assessed at timed intervals with respect to cell number, distribution, viability, metabolic activity, cell cycle, presence of contractile proteins (sarcomeric alpha-actin, troponin I, and tropomyosin), and contractile function in response to electrical stimulation [excitation threshold (ET), maximum capture rate (MCR), response to a gap junctional blocker]. Interstitial flow of culture medium through the central 5-mm-diameter x 1.5-mm-thick region resulted in a physiological density of viable and differentiated, aerobically metabolizing cells, whereas dish culture resulted in constructs with only a 100- to 200-microm-thick surface layer containing viable and differentiated but anaerobically metabolizing cells around an acellular interior. Perfusion resulted in significantly higher numbers of live cells, higher cell viability, and significantly more cells in the S phase compared with dish-grown constructs. In response to electrical stimulation, perfused constructs contracted synchronously, had lower ETs, and recovered their baseline function levels of ET and MCR after treatment with a gap junctional blocker; dish-grown constructs exhibited arrhythmic contractile patterns and failed to recover their baseline MCR levels. PMID- 14551060 TI - Responses of chronically hypoxic rat hearts to ischemia: KATP channel blockade does not abolish increased RV tolerance to ischemia. AB - Chronic hypoxia may precondition the myocardium and protect from ischemia reperfusion damage. We therefore examined the recovery of left and right ventricular function after ischemia and reperfusion (15 min each) in isolated blood-perfused working hearts from normoxic (Norm) and hypoxic (Hypo; 14 days, 10.5% O(2)) adult rats. In addition, the mRNA expression of hypoxia-inducible factor (HIF)-1alpha and the protein expression of endothelial nitric oxide synthase (eNOS) were measured. Postischemic left ventricular function recovered to 66 +/- 6% and 67 +/- 5% of baseline in Norm and Hypo, respectively. In contrast, postischemic right ventricular function was 93 +/- 2% of baseline in Hypo vs. 67 +/- 3% in Norm (P < 0.05). Improved postischemic right ventricular function in Hypo (93 +/- 2% and 96 +/- 2% of baseline) was observed with 95% O(2) or 21% O(2) in the perfusate, and it was not attenuated by glibenclamide (5 and 10 micromol/l) (86 +/- 4% and 106 +/- 6% recovery). HIF-1alpha mRNA and eNOS protein expression were increased in both left and right hypoxic ventricles. In conclusion, postischemic right, but not left, ventricular function was improved by preceding chronic hypoxia. ATP-sensitive K(+) channels are not responsible for the increased right ventricular tolerance to ischemia after chronic hypoxia in adult rat hearts. PMID- 14551061 TI - Secretagogue-stimulated pancreatic secretion is differentially regulated by constitutive NOS isoforms in mice. AB - Nitric oxide (NO) and NO synthase (NOS) play controversial roles in pancreatic secretion. NOS inhibition reduces CCK-stimulated in vivo pancreatic secretion, but it is unclear which NOS isoform is responsible, because NOS inhibitors lack specificity and three NOS isoforms exist: neuronal (nNOS), endothelial (eNOS), and inducible (iNOS). Mice having individual NOS gene deletions were used to clarify the NOS species and cellular interactions influencing pancreatic secretion. In vivo secretion was performed in anesthetized mice by collecting extraduodenal pancreatic duct juice and measuring protein output. Nonselective NOS blockade was induced with N(omega)-nitro-L-arginine (L-NNA; 10 mg/kg). In vivo pancreatic secretion was maximal at 160 pmol.kg(-1).h(-1) CCK octapeptide (CCK-8) and was reduced by NOS blockade (45%) and eNOS deletion (44%). Secretion was unaffected by iNOS deletion but was increased by nNOS deletion (91%). To determine whether the influence of NOS on secretion involved nonacinar events, in vitro CCK-8-stimulated secretion of amylase from isolated acini was studied and found to be unaltered by NOS blockade and eNOS deletion. Influence of NOS on in vivo secretion was further examined with carbachol. Protein secretion, which was maximal at 100 nmol.kg(-1).h(-1) carbachol, was reduced by NOS blockade and eNOS deletion but unaffected by nNOS deletion. NOS blockade by L-NNA had no effect on carbachol-stimulated amylase secretion in vitro. Thus constitutive NOS isoforms can exert opposite effects on in vivo pancreatic secretion. eNOS likely plays a dominant role, because eNOS deletion mimics NOS blockade by inhibiting CCK-8 and carbachol-stimulated secretion, whereas nNOS deletion augments CCK-8 but not carbachol-stimulated secretion. PMID- 14551062 TI - Prescribing and taking medicines. PMID- 14551063 TI - Taking hormone replacement therapy. PMID- 14551064 TI - Is concordance the primrose path to health? PMID- 14551066 TI - Directly observed treatment for tuberculosis. PMID- 14551065 TI - Treatment of tuberculosis. PMID- 14551067 TI - The patient's perspective on medicines in mental illness. PMID- 14551069 TI - Nobel prize in medicine awarded to MRI pioneers. PMID- 14551070 TI - Teacher was refused job because relatives have Huntington's disease. PMID- 14551072 TI - Research should be accessible to all, says Wellcome Trust. PMID- 14551073 TI - Governments failing to invest in education and health. PMID- 14551075 TI - Number of uninsured in the United States increases by 2.4m. PMID- 14551076 TI - FDA chief wants other rich countries to share drug development costs. PMID- 14551077 TI - German doctor is investigated for killing 76 patients with morphine. PMID- 14551080 TI - GPs should encourage more HIV testing, says AIDS foundation. PMID- 14551083 TI - Women lose legal fight to use their frozen embryos. PMID- 14551085 TI - Africa's visionary editor. PMID- 14551088 TI - Parents key to reducing overweight in children. PMID- 14551091 TI - Netherlands postpones smoking ban in restaurants and theatres. PMID- 14551092 TI - British Iraqi doctors set up charity to support Iraq's mental health services. PMID- 14551093 TI - Canada decides to lead the way in exempting AIDS drugs from patent laws. PMID- 14551095 TI - Hong Kong hampered in fight against SARS by lack of cooperation from mainland. PMID- 14551096 TI - Compliance therapy: a randomised controlled trial in schizophrenia. AB - OBJECTIVE: To evaluate the efficacy of "compliance therapy" for improving adherence to prescribed drug treatment among patients with schizophrenia. DESIGN: Randomised controlled trial. SETTING: Urban catchment area psychiatric service. PARTICIPANTS: 94 consecutive admissions of patients with schizophrenia, 56 agreed to participate. INTERVENTION: Compliance therapy and non-specific counselling, each consisting of 5 sessions lasting 30-60 minutes. MAIN OUTCOME MEASURES: Compliance with drug treatment at one year; attitudes to treatment, symptomatology, insight, and quality of life at one year; length of "survival" in the community, bed days, and rehospitalisation rates at two years. RESULTS: Compliance therapy did not confer a major advantage over non-specific therapy in improving compliance at one year (43% (12/28) v 54% (15/28), difference -11% (95% confidence interval -37% to 15%) or in any of the secondary outcome measures symptomatology, attitudes to treatment, insight, global assessment of functioning, and quality of life. CONCLUSION: Compliance therapy may not be of benefit to patients with schizophrenia. Attitudes to treatment at baseline predicted adherence one year later and may be a clinically useful tool. PMID- 14551097 TI - Managing multiple morbidity in mid-life: a qualitative study of attitudes to drug use. AB - OBJECTIVE: To examine attitudes towards drug use among middle aged respondents with high levels of chronic morbidity. DESIGN: Qualitative study with detailed interviews. SETTING: West of Scotland. PARTICIPANTS: 23 men and women aged about 50 years with four or more chronic illnesses. MAIN OUTCOME MEASURE: Participants' feelings about long term use of drugs to manage chronic multiple morbidity. RESULTS: Drugs occupied a central place in the way people managed their comorbidities. Respondents expressed an aversion to taking drugs, despite acknowledging that they depended on drugs to live as "normal" a life as possible. Respondents expressed ambivalence to their drugs in various ways. Firstly, they adopted both regular and more flexible regimens and might adhere to a regular regimen in treating one condition (such as hypertension) while adopting a flexible regimen in relation to others, in response to their experience of symptoms or varying demands of their daily life. Secondly, they expressed reluctance to take drugs, but an inability to be free of them. Thirdly, drugs both facilitated performance of social roles and served as evidence of an inability to perform such roles. CONCLUSIONS: Insight into the considerable tension experienced by people managing complex drug regimens to manage multiple chronic illness may help medical carers to support self care practices among patients and to optimise concordance in their use of prescribed drugs. PMID- 14551098 TI - Does a prescribed treatment match a patient's priorities? PMID- 14551099 TI - Factors involved in deciding to start preventive treatment: qualitative study of clinicians' and lay people's attitudes. AB - OBJECTIVES: To explore the views of clinicians and lay people about the minimum benefit needed to justify drug treatment to prevent heart attacks, and to explore the rationale behind treatment decisions. DESIGN: Qualitative study using semi structured interviews. PARTICIPANTS: 4 general practitioners, 4 practice nurses, and 18 lay people. SETTING: 8 general practices and 6 community settings across Liverpool. RESULTS: Participants varied widely in the minimum acceptable benefits chosen. Most people found the concepts difficult initially, and few appreciated that increased length of treatment should increase absolute benefits. Lay people usually wanted to make decisions for themselves, and clinicians supported this. Participants wanted to consider adverse effects and costs of treatment. Dislike of drug taking was common, and many people preferred lifestyle change to an imperfect treatment. Quality of life and personal views were more important than an individual's age. CONCLUSIONS: Evidence based guidelines make assumptions about people's preferences, and, by using 10 year estimates of risk, inflate the apparent benefits of treatment. It is unlikely that guidelines could incorporate the wide range of people's preferences, and true dialogue is necessary between clinicians and patients before starting long term preventive treatment. PMID- 14551100 TI - A small study, but the results ring true. PMID- 14551101 TI - Changes in use of hormone replacement therapy after the report from the Women's Health Initiative: cross sectional survey of users. PMID- 14551102 TI - Managing haemorrhoids. PMID- 14551103 TI - Assisted conception. II--in vitro fertilisation and intracytoplasmic sperm injection. PMID- 14551104 TI - A wolf in sheep's clothing: a critical look at the ethics of drug taking. PMID- 14551105 TI - Concordance and children's use of medicines. PMID- 14551106 TI - What information do patients need about medicines? Ask the patients--they may want to know more than you think. PMID- 14551108 TI - What information do patients need about medicines? "Doc, tell me what I need to know"--a doctor's perspective. PMID- 14551109 TI - What information do patients need about medicines? Perspectives from the pharmaceutical industry. PMID- 14551110 TI - What information do patients need about medicines? "Doing prescribing": how doctors can be more effective. PMID- 14551111 TI - Women need better information on routine mammography: women's needs inform contents of screening literature. PMID- 14551112 TI - Women need better information on routine mammography: information on expected mortality reduction from attending screening must be correct. PMID- 14551113 TI - Women need better information on routine mammography: understanding of principles of screening must be improved. PMID- 14551114 TI - Women need better information on routine mammography: framing is important in presenting risk information. PMID- 14551115 TI - Separation of anxiety and depressive disorders: normal rules of critical evaluation were presumably suspended. PMID- 14551116 TI - Separation of anxiety and depressive disorders: New tools will lead to more valid classification system. PMID- 14551117 TI - Study of plantar fasciitis treatment is flawed. PMID- 14551118 TI - Separation of anxiety and depressive disorders: maybe pharmaceutical failure has created culture of niche diagnosis. PMID- 14551119 TI - Care and outcomes in young adults with type 1 diabetes: more laser treatment is used in England than the Netherlands. PMID- 14551120 TI - Identifying patients at low risk of bowel cancer: personal or familial risk factors need to be mentioned. PMID- 14551121 TI - Care and outcomes in young adults with type 1 diabetes: services do not usually include psychological care. PMID- 14551122 TI - Tables for predicting survival for preterm births are updated. PMID- 14551123 TI - Time to clarify effect of socioeconomic class on subfertility. PMID- 14551124 TI - Use eggs, not embryos, to derive stem cells. PMID- 14551125 TI - "Egg giving" is trading, not one way process of giving. PMID- 14551126 TI - A career in sexual and reproductive health. PMID- 14551127 TI - Dosh for dummies. PMID- 14551130 TI - The higher professional education scheme for new GPs. PMID- 14551132 TI - Transfer of allogeneic CD62L- memory T cells without graft-versus-host disease. AB - The major challenge in allogeneic hematopoietic cell transplantation is how to transfer allogeneic T-cell immunity without causing graft-versus-host disease (GVHD). Here we report a novel strategy to selectively prevent GVHD by depleting CD62L(+) T cells (naive and a subset of memory T cells). In unprimed mice, CD62L( ) T cells (a subset of memory T cells) failed to proliferate in response to alloantigens (which the mice have never previously encountered) and were unable to induce GVHD in allogeneic hosts. CD62L(-) T cells contributed to T-cell reconstitution by peripheral expansion as well as by promoting T-cell regeneration from bone marrow stem/progenitor cells. CD62L(-) T cells from the animals previously primed with a tumor cell line (BCL1) were able to inhibit the tumor growth in vivo but were unable to induce GVHD in the third-party recipients. This novel technology may allow transfer of allogeneic recall antitumor and antimicrobial immunity without causing GVHD. PMID- 14551133 TI - Treatment of Philadelphia chromosome-positive acute lymphocytic leukemia with hyper-CVAD and imatinib mesylate. AB - Imatinib mesylate, an inhibitor of the Bcr-Abl tyrosine kinase, has modest activity in refractory/relapsed Philadelphia chromosome (Ph)-positive acute lymphocytic leukemia (ALL). Use of concurrent chemotherapy and imatinib mesylate in newly diagnosed Ph-positive ALL was explored. There were 20 patients who received hyper-CVAD (cyclophosphamide, vincristine, Adriamycin, and dexamethasone) and imatinib mesylate followed by imatinib mesylate-based consolidation/maintenance therapy. Of these patients, 11 had de novo disease, 4 were primary failures after induction (without imatinib mesylate), and 5 were in complete remission (CR) after induction (without imatinib mesylate). All 15 patients treated for active disease achieved CR. Within a median of 3.5 months in first CR, 10 patients underwent allogeneic stem cell transplantation (SCT). One patient relapsed after matched related SCT. The other 9 patients remained alive in CR with median follow-up of 12 months after SCT (range, 1+ to 17+ months). Among 10 patients ineligible for (no donor or older age) or refusing allogeneic SCT, 1 patient relapsed after one year. There were 5 patients who remained alive in continuous CR for a median of 20 months (range, 4+ to 24+ months), with 2 older patients dying in CR at 15 and 16 months of comorbid conditions. Molecular CRs were achieved in both groups (SCT or no SCT). Outcome with hyper-CVAD and imatinib mesylate appears better than with prior regimens; continued accrual and longer follow-up of the current cohort is needed. PMID- 14551134 TI - Total correction of hemophilia A mice with canine FVIII using an AAV 8 serotype. AB - Despite the popularity of adeno-associated virus 2 (AAV2) as a vehicle for gene transfer, its efficacy for liver-directed gene therapy in hemophilia A or B has been suboptimal. Here we evaluated AAV serotypes 2, 5, 7, and 8 in gene therapy of factor VIII (FVIII) deficiency in a hemophilia A mouse model and found that AAV8 was superior to the other 3 serotypes. We expressed canine B domain-deleted FVIII cDNA either in a single vector or in 2 separate AAV vectors containing the heavy- and light-chain cDNAs. We also evaluated AAV8 against AAV2 in intraportal and tail vein injections. AAV8 gave 100% correction of plasma FVIII activity irrespective of the vector type or route of administration. PMID- 14551135 TI - Identification of critical amino-acid residues on the erythroid intercellular adhesion molecule-4 (ICAM-4) mediating adhesion to alpha V integrins. AB - Intercellular adhesion molecule-4 (ICAM-4, syn. LW glycoprotein) interacts with the integrins alpha(L)beta(2), alpha(M)beta(2), A(4)beta(1), the alpha(V) family, and alpha(IIb)beta(3). Systematic mutagenesis of surface-exposed residues conserved between human and murine ICAM-4 defined 12 single amino-acid changes that affect the interaction of ICAM-4 with alpha(V) integrins. Mutation of 10 of these residues, 8 of which are spatially close on the surface of the molecule, led to a reduction in adhesion. Moreover, peptides corresponding to regions of ICAM-4 involved in its interaction with alpha(V) integrins inhibited these interactions. The other 2 mutations increased the extent of interaction of ICAM-4 with alpha(V) integrins. These mutations appear to prevent glycosylation of N160, suggesting that changes in glycosylation may modulate ICAM-4-alpha(V) integrin interactions. The region of ICAM-4 identified as the binding site for alpha(V) integrins is adjacent to the binding sites for alpha(L)beta(2) and alpha(M)beta(2). Selective binding of ICAM-4 to different integrins may be important for a variety of normal red cell functions and also relevant to the pathology of thrombotic disorders and vasoocclusive events in sickle cell disease. Our findings suggest the feasibility of developing selective inhibitors of ICAM-4-integrin adhesion of therapeutic value in these diseases. PMID- 14551136 TI - A novel SHP-1/Grb2-dependent mechanism of negative regulation of cytokine receptor signaling: contribution of SHP-1 C-terminal tyrosines in cytokine signaling. AB - SHP-1, an src homology 2 (SH2) domain containing protein tyrosine phosphatase, functions as a negative regulator of signaling downstream of cytokine receptors, receptor tyrosine kinases and receptor complexes of the immune system. Dephosphorylation of receptors and/or receptor-associated kinases has been described as the mechanism for the function of SHP-1. Here we demonstrate a novel mechanism by which SHP-1 down-regulates the Janus kinase-2 (Jak2)/signal transducer and activator of transcription-5 (Stat5) pathway downstream of the prolactin receptor (PRLR) and the erythropoietin receptor (EPOR) in a catalytic activity-independent manner. Structural/functional analysis of SHP-1 defined the C-terminal tyrosine residues (Y278, Y303, Y538, Y566) within growth factor receptor-bound protein 2 (Grb-2) binding motif to be responsible for delivering the inhibitory effects. Our results further indicate that these tyrosine residues, via recruitment of the adaptor protein Grb-2, are required for targeting the inhibitory protein suppressor of cytokine signaling-1 (SOCS-1) to Jak2 kinase. Finally, loss of SOCS-1 expression in SOCS-1(-/-) mouse embryonic fibroblast (MEF) cells led to attenuation in SHP-1 function to down-regulate PRL induced Stat5 activation. All together, our results indicate that SHP-1 inhibits PRLR and EPOR signaling by recruitment and targeting of SOCS-1 to Jak2, highlighting a new mechanism of SHP-1 regulation of cytokine-receptor signaling. PMID- 14551137 TI - LAD-III, a leukocyte adhesion deficiency syndrome associated with defective Rap1 activation and impaired stabilization of integrin bonds. AB - Recently, we reported a rare leukocyte adhesion deficiency (LAD) associated with severe defects in integrin activation by chemokine signals, despite normal ligand binding of leukocyte integrins.(1) We now report that the small GTPase, Rap1, a key regulator of inside-out integrin activation is abnormally regulated in LAD Epstein-Barr virus (EBV) lymphocyte cells. Both constitutive and chemokine triggered activation of Rap1 were abolished in LAD lymphocytes despite normal chemokine signaling. Nevertheless, Rap1 expression and activation by phorbol esters were intact, ruling out an LAD defect in Rap1 guanosine triphosphate (GTP) loading. The very late antigen 4 (VLA-4) integrin abnormally tethered LAD EBV lymphocytes to its ligand vascular cell adhesion molecule 1 (VCAM-1) under shear flow due to impaired generation of high-avidity contacts despite normal ligand binding and intact avidity to surface-bound anti-VLA-4 monoclonal antibody (mAb). Thus, a defect in constitutive Rap1 activation results in an inability of ligand occupied integrins to generate high-avidity binding to ligand under shear flow. This is a first report of an inherited Rap1 activation defect associated with a pathologic disorder in leukocyte integrin function, we herein term it "LAD-III." PMID- 14551138 TI - 17-Allylamino-17-demethoxygeldanamycin (17-AAG) is effective in down-regulating mutated, constitutively activated KIT protein in human mast cells. AB - Mutations in the proto-oncogene c-kit cause constitutive kinase activity of its product, KIT protein, and are associated with human mastocytosis and gastrointestinal stromal tumors (GISTs). Although currently available tyrosine kinase inhibitors are effective in the treatment of GISTs, there has been limited success in the treatment of mastocytosis. 17-Allylamino-17-demethoxygeldanamycin (17-AAG), a benzoquinoid ansamycin antibiotic, which binds to heat shock protein 90 (hsp90) causes destabilization of various hsp90-dependent kinases important in oncogenesis. Treatment with 17-AAG of the mast cell line HMC-1.2, harboring the Asp816Val and Val560Gly KIT mutations, and the cell line HMC-1.1, harboring a single Val560Gly mutation, causes both the level and activity of KIT and downstream signaling molecules AKT and STAT3 to be down-regulated following drug exposure. These data were validated using Cos-7 cells transfected with wild-type and mutated KIT. 17-AAG promotes cell death of both HMC mast cell lines. In addition, neoplastic mast cells isolated from patients with mastocytosis, incubated with 17-AAG ex vivo, are selectively sensitive to the drug compared to the mononuclear fraction. These data provide compelling evidence that 17-AAG may be effective in the treatment of c-kit-related diseases including mastocytosis, GISTs, mast cell leukemia, subtypes of acute myelogenous leukemia, and testicular cancer. PMID- 14551139 TI - Identification and characterization of EBP, a novel EEN binding protein that inhibits Ras signaling and is recruited into the nucleus by the MLL-EEN fusion protein. AB - The chimeric MLL-EEN fusion protein is created as a result of chromosomal translocation t(11;19)(q23;p13). EEN, an Src homology 3 (SH3) domain-containing protein in the endophilin family, has been implicated in endocytosis, although little is known about its role in leukemogenesis mediated by the MLL-EEN fusion protein. In this study, we have identified and characterized EBP, a novel EEN binding protein that interacts with the SH3 domain of EEN through a proline-rich motif PPERP. EBP is a ubiquitous protein that is normally expressed in the cytoplasm but is recruited to the nucleus by MLL-EEN with a punctate localization pattern characteristic of the MLL chimeric proteins. EBP interacts simultaneously with EEN and Sos, a guanine-nucleotide exchange factor for Ras. Coexpressoin of EBP with EEN leads to suppression of Ras-induced cellular transformation and Ras mediated activation of Elk-1. Taken together, our findings suggest a new mechanism for MLL-EEN-mediated leukemogenesis in which MLL-EEN interferes with the Ras-suppressing activities of EBP through direct interaction. PMID- 14551140 TI - Effect of a single injection of humanized anti-CD154 monoclonal antibody on the platelet-specific autoimmune response in patients with immune thrombocytopenic purpura. AB - Blockade of the CD40/CD154 signal is a potential immunomodulatory strategy for T cell-mediated diseases. As a part of a phase 1, multicenter, dose-escalating trial of humanized monoclonal antibody to CD154 (IDEC-131/E6040) in patients with refractory immune thrombocytopenic purpura (ITP), the autoimmune response to glycoprotein IIb/IIIa (GPIIb/IIIa) was evaluated at successive time points. Five patients each were given a single infusion of 1, 2, 5, or 10 mg/kg IDEC-131/E6040 and followed for 3 months. All adverse events were mild, and there were no severe infections or thromboembolic events. No increase in platelet count was observed in patients treated at 1, 2, or 5 mg/kg, but an increase was observed in 3 patients treated at 10 mg/kg. In only the patients treated at 5 or 10 mg/kg, the frequency of B cells producing anti-GPIIb/IIIa antibodies, GPIIb/IIIa-induced T cell proliferation, and anti-GPIIb/IIIa antibody production by antigen-dependent T-B-cell collaboration were all suppressed in parallel after the treatment, with a slow return to baseline. In contrast, T-cell response to an irrelevant antigen was not affected. These findings suggest that CD40/CD154 blockade therapy is potentially effective for refractory ITP, through selective suppression of autoreactive T and B cells to platelet antigens. PMID- 14551141 TI - Clofarabine, a novel nucleoside analog, is active in pediatric patients with advanced leukemia. AB - Despite progress in leukemia therapy, most children who experience relapse have a dismal prognosis. New, effective approaches are needed. We conducted a phase 1 study of a novel nucleoside analog, clofarabine, in pediatric patients with refractory and relapsed leukemia. Clofarabine was infused intravenously over 1 hour each day for 5 days. Six dose levels, between 11.25 and 70 mg/m(2) per day for 5 days, were studied in 25 patients. A modified 3 + 3 phase 1 design was followed with 30% dose escalation until the dose-limiting toxicity (DLT) was defined. The maximum tolerated dose (MTD) was 52 mg/m(2) per day for 5 days. At the end of infusion at MTD, clofarabine triphosphate levels in leukemia blasts varied between 6 microM and 19 microM, which resulted in complete and sustained inhibition of DNA synthesis. The DLT was reversible hepatotoxicity and skin rash at 70 mg/m(2) per day for 5 days. Twenty-five patients were treated. Five patients achieved complete remission (CR), and 3 achieved partial remission (PR), for an overall response rate of 32%. Clofarabine is well tolerated and shows significant antileukemic activity in heavily pretreated children. Multicenter phase 2 trials in pediatric acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML) are ongoing. PMID- 14551142 TI - ETO protein of t(8;21) AML is a corepressor for Bcl-6 B-cell lymphoma oncoprotein. AB - The multiplicity of transcription factors involved in hematologic malignancies suggests a complicated scenario in which many different molecular mechanisms lead to malignant transformation. We hypothesized that some of these proteins might physically and functionally interact and thus mechanistically link different diseases. The ETO protein of t(8;21) acute myeloid leukemia (AML) is an excellent candidate as a common factor because it is normally expressed in human hematopoietic cells, it binds to histone deacetylases (HDACs), and it interacts with the PLZF protein of t(11;17) acute promyelocytic leukemia. To determine whether ETO functionally links a broader range of disease entities, we asked whether ETO forms a complex with the Bcl-6 oncoprotein of B-cell lymphomas. We found that ETO and Bcl-6 are coexpressed in normal and malignant lymphoid tissue, where they interact and colocalize in nuclear speckles. ETO binds to the fourth zinc finger of Bcl-6, enhances Bcl-6 repression of artificial and endogenous genes in an HDAC-dependent manner, and forms a complex with Bcl-6 on the promoters of its endogenous target genes in B-cell lymphoma cells. Therefore, ETO is a bona fide corepressor that links the transcriptional pathogenesis of acute leukemias and B-cell lymphomas and offers a compelling target for transcriptional therapy of hematologic malignancies. PMID- 14551143 TI - Antibody specificity controls in vivo effector mechanisms of anti-CD20 reagents. AB - Despite the success of anti-CD20 monoclonal antibody (mAb) in the treatment of lymphoma, there remains considerable uncertainty about their mechanism(s) of action. Here, we show that certain of these reagents (rituximab and 1F5), which redistribute CD20 into membrane rafts, are bound efficiently by C1q, deposit C3b, and result in complement-dependent cytotoxicity (CDC). This activity is important in vivo, because complement depletion using cobra venom factor (CVF) markedly reduced the efficacy of rituximab and 1F5 in 2 lymphoma xenograft models. However, complement depletion had no effect on the potent therapeutic activity of B1, a mAb that does not redistribute CD20 into membrane rafts, bind C1q, or cause efficient CDC. Equivalent immunotherapy also occurred in the presence or absence of natural killer (NK) cells. Perhaps most surprising was the observation that F(ab')2 fragments of B1 but not 1F5 were able to provide substantial immunotherapy, indicating that non-Fc-dependent mechanisms are involved with B1. In accordance with this, B1 was shown to induce much higher levels of apoptosis than rituximab and 1F5. Thus, although complement is important for the action of rituximab and 1F5, this is not so for B1, which more likely functions through its ability to signal apoptosis. PMID- 14551144 TI - Role of adenosine receptors in regulating chemotaxis and cytokine production of plasmacytoid dendritic cells. AB - Plasmacytoid dendritic cells (PDCs) are potent regulators of immune function and the major source of type I interferon (IFN) following viral infection. PDCs are found at sites of inflammation in allergic reactions, autoimmune disorders, and cancer, but the mechanisms leading to the recruitment of PDCs to these sites remain elusive. During inflammation, adenosine is released and functions as a signaling molecule via adenosine receptors. This study analyzes adenosine receptor expression and function in human PDCs. Adenosine was found to be a potent chemotactic stimulus for immature PDCs via an A(1) receptor-mediated mechanism. The migratory response toward adenosine was comparable to that seen with CXCL12 (stromal-derived factor-1 alpha [SDF-1 alpha), the most potent chemotactic stimulus identified thus far for immature PDCs. Upon maturation, PDCs down-regulate the A(1) receptor, resulting in a loss of migratory function. In contrast, mature PDCs up-regulate the A(2a) receptor, which is positively coupled to adenylyl cyclase and has been implicated in the down-regulation of DC cytokine producing capacity. We show that in mature PDCs adenosine reduces interleukin-6 (IL-6), IL-12, and IFN-alpha production in response to CpG oligodeoxynucleotides (ODN). These findings indicate that adenosine may play a dual role in PDC mediated immunity by initially recruiting immature PDCs to sites of inflammation and by subsequently limiting the extent of the inflammatory response induced by mature PDCs by inhibiting their cytokine-producing capacity. PMID- 14551145 TI - Mantle cell lymphoma with t(11;14) and unmutated or mutated VH genes expresses AID and undergoes isotype switch events. AB - Isotype switch commonly follows onset of somatic hypermutation in the germinal center (GC), with activation-induced cytidine deaminase (AID) as a prerequisite. Mantle cell lymphoma (MCL) with t(11;14) includes a subset with unmutated (UM) and a minor subset with mutated (MUT) VH genes. Here, we investigated whether switch events and AID expression occur in MCL. In 4 of 6 UM and 4 of 7 MUT MCLs, alternative tumor-derived Cgamma,alpha,epsilon transcripts were identified. AID transcripts, including a splice variant, were common to both subsets. AID expression correlated with switch in 8 of 8 cases, but in 3 of 5 cases it occurred with switch absent. Circle transcripts (Igamma-Cmicro/Ialpha-Cmu) were identified in 5 of 7 evaluated cases. In 1 of 12 cases, 12% of tumor cells expressed immunoglobulin L-restricted surface IgA. Ongoing switch recombination events appear to be a feature of MCL, likely restricted to a minor tumor subpopulation, with occasional variant sIg expression. UM MCLs implicate origins from pre-GC B cells and reveal switch events at ectopic sites. PMID- 14551146 TI - Protein 4.2 is critical to CD47-membrane skeleton attachment in human red cells. AB - The reduction in expression of the integral membrane protein CD47 in human red blood cells (RBCs) deficient in protein 4.2 suggests that protein 4.2 may mediate a linkage of CD47 to the membrane skeleton. We compared the fractions of membrane skeleton-attached CD47, Rh-associated glycoprotein (RhAG), Rh, and band 3 in normal and protein 4.2-deficient cells using fluorescence-imaged microdeformation. We found that CD47 attachment decreases from 55% in normal cells to 25% to 35% in 4.2-deficient cells. RhAG, which has been shown to have no significant variation in expression among the cells studied, shows a significant decrease in membrane skeleton attachment in 4.2-deficient cells from 60% to 40%. Both Rh and band 3, which have also been shown to have no change in expression, show a smaller decrease from 75% attached in normal RBCs to 55% attached in 4.2 deficient cells. In normal cells, Rh phenotype influences CD47 expression but not the level of membrane skeleton attachment of CD47. In contrast, the results indicate that protein 4.2 strongly influences CD47 levels as well as the extent of membrane skeleton attachment in the RBC, whereas protein 4.2 affects membrane skeletal attachment of RhAG, Rh, and band 3 to a lesser extent. PMID- 14551147 TI - Effect of second- and third-generation oral contraceptives on the protein C system in the absence or presence of the factor VLeiden mutation: a randomized trial. AB - A plausible mechanism to explain thrombotic risk differences associated with the use of second- and third-generation oral contraceptives (OCs), particularly in carriers of factor V(Leiden), is still lacking. In a double-blind trial, 51 women without and 35 women with factor V(Leiden) were randomized to either a second- (30 microg ethinylestradiol/150 microg levonorgestrel) or third- (30 microg ethinylestradiol/150 microg desogestrel) generation OC. After 2 cycles of use and a wash-out of 2 cycles, the participants continued with the corresponding progestagen-only preparation. Hemostatic variables that probe the activity of the anticoagulant protein C system were determined. Compared with levonorgestrel, desogestrel-containing OCs significantly decreased protein S and increased activated protein C (APC) resistance in both groups. OCs with desogestrel had the most pronounced effects in carriers of factor V(Leiden). Progestagen-only preparations caused changes of anticoagulant parameters opposite to those of combined OCs, which in a number of cases were more pronounced with levonorgestrel. Our data show that progestagens in combined OCs counteract the thrombotic effect of the estrogen component. The higher thrombotic risk associated with third-generation OCs compared with second-generation OCs may be explained by the fact that desogestrel appeared less antithrombotic than levonorgestrel, especially in women with factor V(Leiden). PMID- 14551148 TI - Prior chemotherapy and allograft CD34+ dose impact donor engraftment following nonmyeloablative allogeneic stem cell transplantation in patients with solid tumors. AB - Significant engraftment variability occurs among patients following nonmyeloablative hematopoietic cell transplantation. We analyzed the impact of multiple factors on donor myeloid and T-cell engraftment in 36 patients with metastatic tumors undergoing cyclophosphamide/fludarabine-based conditioning. Higher CD34(+) doses facilitated donor myeloid engraftment, while prior chemotherapy exposure facilitated both donor myeloid and T-cell engraftment. At day 30, median donor T-cell and myeloid chimerism was 98% and 76%, respectively, in those patients with prior chemotherapy versus 88% (P =.008) and 26% (P <.0001) in chemotherapy-naive patients. Donor myeloid chimerism at day 45 was predicted by prior chemotherapy exposure and the log(10) of the CD34(+) dose (adjusted coefficient of determination [R(2)] =.47; P <.0001), while chemotherapy alone impacted donor T-cell engraftment. Patients with prior chemotherapy were more likely to develop acute grades II to IV graft-versus-host disease (GVHD; 8/18) compared with chemotherapy-naive patients (2/18; P =.031). Thus, tailoring the intensity of nonmyeloablative conditioning based on prior chemotherapy exposure is an important consideration in trial design. PMID- 14551149 TI - Highly efficient expression of transgenic proteins by naked DNA-transfected dendritic cells through terminal differentiation. AB - Dendritic cells (DCs) play a key role in the induction and control of immunity. Genetic engineering of DCs is a promising approach for the development of a broad range of immunomodulatory strategies, for purposes ranging from genetic immunization to tolerance induction. The development of DC-based immunotherapies is limited by the inability to efficiently transfect DCs using naked DNA. Here we demonstrate that after plasmid DNA delivery, the transgene expression level controlled by the human immediate-early cytomegalovirus promoter (hIE-CMVp) is higher in mature DCs than in immature DCs and is further increased after terminal differentiation of DCs by agonist anti-CD40 monoclonal antibody (mAb) or after DC interaction with CD4(+) T cells. CD40 signaling of DCs resulted in nuclear translocation of the transcription factors nuclear factor-kappaB (NF-kappaB), activator of protein-1 (AP-1), and cyclic adenosine monophosphate (cAMP) responsive element, necessary for the activation of hIE-CMVp. Transgene expression by DCs diminished after the inhibition of these transcription factors or the blockade of adhesion molecules involved in the DC-T-cell synapse. Importantly, CD40 signaling of DCs results in the highly efficient expression and presentation of transgenic antigens and the induction of "in vivo" cytotoxic T cell (CTL) responses specific for transgenic antigen peptides, demonstrating the functional potential of genetically engineered DCs. PMID- 14551150 TI - Critical role of interleukin-1beta for transcriptional regulation of endothelial 6-pyruvoyltetrahydropterin synthase. AB - OBJECTIVE: Synthesis of tetrahydrobiopterin (BH4), an essential cofactor for nitric oxide synthases, is strongly induced on immunostimulation in vascular endothelial cells (VECs). Expression of GTP cyclohydrolase I (GTPCH), the first enzyme in BH4 biosynthesis, is regulated by cytokines and considered rate limiting. Herein we investigated the molecular mechanism and relevance of cytokine-dependent regulation of 6-pyruvoyltetrahydropterin synthase (PTPS), the second enzyme in BH4 synthesis, in human coronary artery endothelial cells (HCAECs). METHODS AND RESULTS: Real-time polymerase chain reaction revealed a 4 fold induction of PTPS and a 300-fold induction of GTPCH expression by interleukin (IL)-1beta/tumor necrosis factor-alpha/interferon-gamma, mainly through de novo transcription. On immunostimulation, PTPS became rate-limiting. Importantly, IL-1beta induced PTPS rather than GTPCH. As a result, IL-1beta contributed significantly to the amount of BH4 produced (+40%) but concomitantly reduced the accumulation of the GTPCH intermediate, 7,8-dihydroneopterin triphosphate (-50%). CONCLUSIONS: Our data show that PTPS induction is necessary for optimized BH4 synthesis in cytokine-stimulated HCAECs and point to IL-1beta as a leading cytokine in this process. PMID- 14551151 TI - Adiponectin and metabolic syndrome. AB - In this review article, the crucial roles of adipocytes in the development of so called metabolic syndrome and vascular disease are reviewed, focusing on adipocyte-derived bioactive substances, adipocytokines. Recent progress in adipocyte biology shows that adipocytes are not merely energy-storing cells but that they secrete a variety of hormones cytokines, growth factors, and other bioactive substances. To search for novel adipocytokines by the large-scale random sequence analysis of expressed genes in adipocytes, we identified an adipose-specific collagen-like molecule, adiponectin. This novel adipocytokine has plural biofunctions, such as antidiabetic, antiatherosclerotic, and antiinflammatory functions. Adiponectin plasma levels decrease with the accumulation of visceral adipose tissue. In this review, we discuss the link of adiponectin to visceral adiposity, insulin resistance, and vascular diseases. PMID- 14551152 TI - Nuclear localization of endothelin-converting enzyme-1: subisoform specificity. AB - OBJECTIVE: The biosynthesis of endothelin-1 (ET-1), the most potent vasoconstrictor with mitogenic properties, involves the processing of intermediate protein big ET-1 by a unique metalloprotease, endothelin-converting enzyme-1 (ECE-1). ECE-1 has 4 subisoforms that possess the same catalytic properties but different localization patterns on the plasma membrane and cytosol. We investigated the trafficking of ECE-1 subisoforms using green fluorescent protein-tagged recombinant enzymes in target and nontarget cells. METHODS AND RESULTS: ECE-1 localization was studied using confocal microscopy, which provides evidence for the first time that both ET-1 and ECE-1a are also found in the nuclear compartment in transiently transfected cells as well as in native endothelial cells that endogenously possess the ET system. In cells maintained in high-glucose medium, ECE-1a-specific staining shifted from plasma membrane to intracellular compartments. ECE-1b subisoform, however, is mainly in the cytosolic compartment, indicating a subisoform specificity for nuclear localization. CONCLUSIONS: Our findings define a novel localization pattern for the ET system, which may be differentially regulated under pathophysiological conditions. PMID- 14551153 TI - Insulin-like growth factor-1 receptor activation inhibits oxidized LDL-induced cytochrome C release and apoptosis via the phosphatidylinositol 3 kinase/Akt signaling pathway. AB - OBJECTIVE: We have shown previously that oxidized LDL decreases insulin-like growth factor-1 (IGF-1) and IGF-1 receptor expression in vascular smooth muscle cells and that IGF-1 and IGF-1 receptor expression are reduced in the deep intima of early atherosclerotic lesions. Because oxidized LDL is potentially important for the depletion of vascular smooth muscle cells contributing to plaque destabilization, we studied the role of IGF-1 in oxidized LDL-induced apoptosis. METHODS AND RESULTS: We provide evidence that oxidized LDL-induced apoptosis is caused by decreased mitochondrial membrane potential and increased cytochrome C release in human aortic vascular smooth muscle cells. Overexpression of the IGF-1 receptor by using an adenovirus completely abrogated these effects. The antiapoptotic function of the IGF-1 receptor was associated with increased Akt kinase activity and increased expression of phosphorylated Bad. Moreover, a dominant-negative p85 phosphatidylinositol 3-kinase adenovirus blocked the capacity of the IGF-1 receptor to prevent oxidized LDL-induced apoptosis. CONCLUSIONS: Our data demonstrate that IGF-1 receptor activation inhibits oxidized LDL-induced cytochrome C release and apoptosis through the phosphatidylinositol 3-kinase/Akt signaling pathway and suggest that genetic or pharmacological activation of the IGF-1 receptor may be a useful strategy to stabilize atherosclerotic plaques. PMID- 14551154 TI - Thrombin and phenotypic modulation of the endothelium. AB - Thrombin signaling in the endothelium is linked to multiple phenotypic changes, including alterations in permeability, vasomotor tone, and leukocyte trafficking. The thrombin signal is transduced, at least in part, at the level of gene transcription. In this review, we focus on the role of thrombin signaling and transcriptional networks in mediating downstream gene expression and endothelial phenotype. In addition, we report the results of DNA microarrays in control and thrombin-treated endothelial cells. We conclude that (1) thrombin induces the upregulation and downregulation of multiple genes in the endothelium, (2) thrombin-mediated gene expression involves a multitude of transcription factors, and (3) future breakthroughs in the field will depend on a better understanding of the spatial and temporal dynamics of these transcriptional networks. PMID- 14551155 TI - Linkage and association between distinct variants of the APOA1/C3/A4/A5 gene cluster and familial combined hyperlipidemia. AB - OBJECTIVE: Combined hyperlipidemia is a common disorder, characterized by a highly atherogenic lipoprotein profile and a substantially increased risk of coronary heart disease. The purpose of this study was to establish whether variations of apolipoprotein A5 (APOA5), a newly discovered gene of lipid metabolism located 30 kbp downstream of the APOA1/C3/A4 gene cluster, contributes to the transmission of familial combined hyperlipidemia (FCHL). METHODS AND RESULTS: We performed linkage and association tests on 128 families. Two independent alleles, APOA5c.56G and APOC3c.386G, of the APOA1/C3/A4/A5 gene cluster were overtransmitted in FCHL (P=0.004 and 0.007, respectively). This was paired with reduced transmission of the common APOA1/C3/A4/A5 haplotype (frequency 0.4461) to affected subjects (P=0.012). The APOA5c.56G genotype accounted for 7.3% to 13.8% of the variance in plasma triglyceride levels in probands (P<0.004). The APOC3c.386G genotypes accounted for 4.4% to 5.1% of the variance in triglyceride levels in FCHL spouses (P<0.007), suggesting that this allele marks a FCHL quantitative trait as well as representing a susceptibility locus for the condition. CONCLUSIONS: A combined linkage and association analysis establishes that variation at the APOA1/C3/A4/A5 gene cluster contributes to FCHL transmission in a substantial proportion of northern European families. PMID- 14551156 TI - Anoikis in the cardiovascular system: known and unknown extracellular mediators. AB - Anoikis is defined as programmed cell death induced by the loss of cell/matrix interactions. Adhesion to structural glycoproteins of the extracellular matrix is necessary for survival of the differentiated adherent cells in the cardiovascular system, including endothelial cells, smooth muscle cells, fibroblasts, and cardiac myocytes. Adhesion is also a key factor for the differentiation of mesenchymal stem cells. In particular, fibronectin is considered a factor of survival and differentiation for many adherent cells. Adhesion generates cell tensional integrity (tensegrity) and repression of apoptotic signals, whereas detachment has the opposite effect. Anoikis plays a physiological role by regulating cell homeostasis in tissues. However, anoikis can also be involved in pathological processes, as illustrated by the resistance to anoikis in cancer and its enhancement in degenerative tissue remodeling. Extracellular mediators of anoikis include matrix retraction, leading to loss of tensegrity in fibroblasts, pharmacological disengagement of integrins by RGD-like peptides and fragments of fibronectin, and focal adhesion disassembly by fragments of thrombospondin, plasminogen activator-1, and high-molecular-weight kininogen. In addition to binding of the RGD peptide by integrins, the engagement of the heparin binding sites of adhesive glycoproteins with glycosaminoglycans on the cell surface is also involved in the prevention of cell detachment-induced apoptosis. Proteases able to degrade adhesive glycoproteins, such as fibronectin, induce anoikis of vascular adherent cells. Active proteases can either be secreted directly by inflammatory cells, as elastase and cathepsin G by polymorphonuclear leukocytes, chymase and tryptase by mast cells, and granzymes by lymphocytes, or generated from circulating zymogens by activation in close contact with the cells. This is the case for the pericellular conversion of plasminogen to plasmin, which degrades fibronectin and induces anoikis of smooth muscle cells. Involvement of proteases has also been proposed in the apoptotic response of cultured adherent cells to serum starvation. Anoikis is probably involved in pathological remodeling of cardiovascular tissues, including cardiac myocyte detachment in heart failure, deendothelialization and plaque rupture in atherosclerosis, and smooth muscle cell disappearance in aneurysms and varicose veins. The absence of cell adhesion and growth resulting from cleavage of adhesive proteins also represents a major impediment to cellular healing, including the absence of cell recolonization of proteolytically injured tissue and the low efficacy of cell transplantation. However, the exact role of anoikis in cardiovascular pathologies remains to be further defined. PMID- 14551157 TI - Matrix metalloproteinase-2 and -9 differentially regulate smooth muscle cell migration and cell-mediated collagen organization. AB - OBJECTIVE: Smooth muscle cells (SMCs) produce both matrix metalloproteinase (MMP) 2 and MMP-9, enzymes with similar in vitro matrix degrading abilities. We compared the specific contributions of these enzymes to SMC-matrix interactions in vitro and in vivo. METHODS AND RESULTS: Using genetic models of deficiency, we investigated MMP-2 and MMP-9 roles in SMC migration in vivo in the formation of intimal hyperplasia and in vitro. In addition, we investigated potential effects of MMP-2 and MMP-9 genetic deficiency on compaction and assembly of collagen by SMCs. CONCLUSIONS: MMP-2 and MMP-9 genetic deficiency decreased by 81% and 65%, respectively (P<0.01), SMC invasion in vitro and decreased formation of intimal hyperplasia in vivo (P<0.01). However, we found that MMP-9, but not MMP-2, was necessary for organization of collagen by SMCs. Likewise, we found that MMP-9 deficiency resulted in a 50% reduction of SMC attachment to gelatin (P<0.01), indicating that SMCs may use MMP-9 as a bridge between the cell surface and matrix. Furthermore, we found that the hyaluronan receptor, CD44, assists in attachment and utilization of MMP-9 by SMCs. Understanding the specific roles of these MMPs, generally thought to be similar, could improve the design of therapeutic interventions aimed at controlling vascular remodeling. PMID- 14551158 TI - Sustained expansion and transgene expression of coagulation factor VIII transduced cord blood-derived endothelial progenitor cells. AB - OBJECTIVE: Although hemophilia A seems particularly suitable for gene therapy because even low amounts of plasma coagulation factor VIII (FVIII) provide a significant clinical benefit to the patients, the ideal target cell for recombinant FVIII expression and gene therapy approaches remains to be identified. In this study, we tested the capacity of cord blood-derived endothelial progenitor cells (CBECs) for FVIII expression on stable lentiviral transduction. METHODS AND RESULTS: CD34+ endothelial progenitor cells (EPCs) from cord blood were differentiated into CBECs. Endothelial phenotype was characterized, and lentiviral transduction of early-passage CBECs with a vector encoding FVIII and EGFP did not alter their functional properties and proliferative potential. CBEC could be expanded by 5 to 9 orders of magnitude, thus allowing the expansion of up to 10(15) FVIII-secreting CBECs, starting from as little as 10(6) CD34+ cells. CBECs proved to be highly suitable for FVIII secretion, with 0.35 to 0.39 IU FVIII:C/5x10(4) cells per 48 hours (7.0 to 7.8 IU FVIII:C/10(6) cells per 48 hours), which remained stable over the expansion period. CONCLUSIONS: Our data indicate that CBECs are attractive target cells for inherited coagulation disorders such as hemophilia A, which on lentiviral transduction can be readily expanded to large numbers of transplantable gene modified cells in vitro. PMID- 14551159 TI - Apolipoprotein D inhibits platelet-derived growth factor-BB-induced vascular smooth muscle cell proliferated by preventing translocation of phosphorylated extracellular signal regulated kinase 1/2 to the nucleus. AB - OBJECTIVE: Elevated apolipoprotein D (apoD) levels are associated with reduced proliferation of cancer cells. We therefore investigated whether apoD, which occurs free or associated with HDL, suppresses vascular smooth muscle cell (VSMC) proliferation, which is related to the pathobiology of disease. METHODS AND RESULTS: Intense immunoreactivity for apoD was observed in human atherosclerotic plaque but not in normal coronary artery. However, an increase in apoD mRNA was seen in quiescent relative to proliferating fetal lamb aortic VSMCs, and in the rat aortic VSMC line (A10), we demonstrated uptake of apoD from serum. Stable transfection of apoD in A10 cells in the absence of serum did not influence VSMC proliferation assessed by [3H]-thymidine incorporation. ApoD, administered at a dose of 100 ng/mL, completely inhibited basal as well as platelet-derived growth factor (PDGF)-BB-induced VSMC proliferation (P<0.01) but had no effect on fibroblast growth factor-induced VSMC proliferation. ApoD did not suppress PDGF BB or fibroblast growth factor-2-induced phosphorylation of extracellular signal regulated kinase (ERK) 1/2 but selectively inhibited PDGF-BB-mediated ERK1/2 nuclear translocation. CONCLUSIONS: Our data suggest that apoD selectively modulates the proliferative response of VSMC to growth factors by a mechanism related to nuclear translocation of ERK1/2. PMID- 14551160 TI - Macrolides inhibit epithelial cell-mediated neutrophil survival by modulating granulocyte macrophage colony-stimulating factor release. AB - Macrolides have been shown to be effective in treating diffuse panbronchiolitis (DPB), although the precise modes of action remain unclear. At sites of airway inflammation, respiratory epithelium is considered an active participant in regulating neutrophil survival. We therefore examined the effect of erythromycin, clarithromycin, azithromycin, and josamycin on both neutrophil survival and on epithelial-derived factors, which influence neutrophil longevity. Media conditioned with transiently tumor necrosis factor (TNF)-alpha-stimulated A549 human airway epithelial cells prolonged neutrophil survival compared with control media. The presence of dexamethasone during neutrophil culture led to further prolongation of neutrophil survival. In contrast, none of the tested macrolides modulated neutrophil survival, suggesting a lack of direct effect of these drugs. On the other hand, pretreatment of TNF-alpha-stimulated A549 cells by erythromycin, clarithromycin, azithromycin, or dexamethasone, but not josamycin, decreased the neutrophil survival-enhancing effects in a dose-dependent manner. Neutralizing antibodies to granulocyte macrophage colony-stimulating factor (GM CSF) dampened the prolonged neutrophil survival observed in TNF-alpha-stimulated A549 conditioned media. Erythromycin, clarithromycin, azithromycin, and dexamethasone inhibited TNF-alpha-induced GM-CSF expression in A549 cells at both the protein and messenger RNA levels. These results suggest that macrolides inhibit epithelial cell-mediated neutrophil survival by modulating GM-CSF release, which may, at least in part, explain the effectiveness of this family of drugs on DPB. PMID- 14551161 TI - Mesothelial differentiation as reflected by differential gene expression. AB - Human mesothelial cells obtained from benign effusions retain their proliferative capacity and grow uniformly either with a fibroblastic or epithelioid morphology in vitro. These cultures therefore provide a model for the process of mesothelial differentiation in vivo. To study this differentiation, we isolated differentially expressed genes obtained by suppression subtractive hybridization. Of the nine genes found to be overexpressed in fibroblastic mesothelial cells, three are matrix-associated (integrin alpha5, collagen binding protein 2, human cartilage glycoprotein 39), whereas the others are associated with a proliferative cell type (14-3-3 epsilon, plexin B2, N33, and three genes encoding ribosomal elements). Seven of the eight genes upregulated in the epithelioid phenotype are related rather to specialized functions, such as metabolism (aldose reductase, lecithin:cholesterol acyltransferase, ATPase 6), cytoskeletal composition (cytokeratins 7 and 8), and regulation of differentiation (granulin, annexin II). Immunohistochemistry with available antibodies to six of the differentially expressed gene products confirmed the differences also in pleural tissues, where submesothelial cells displayed the fibroblastic markers, whereas surface cells displayed the epithelioid markers. In summary, this approach revealed a pattern of genes coordinately regulated during mesothelial differentiation and suggests that mesothelium may regenerate also by recruiting cells from the submesothelial layer. Some of the gene products may also be useful markers for differentiation and activation in serosal tissues. PMID- 14551162 TI - Human alveolar wall fibroblasts directly link epithelial type 2 cells to capillary endothelium. AB - Alveolar wall fibroblasts directly link type 2 (T2) pneumocytes to capillary endothelium through apertures in their respective basal laminae in rabbit lung. These fibroblasts provide a bridge from the capillary to the airway lumen along which leukocytes may migrate without disrupting extracellular matrix. Normal human lungs were examined by transmission electron microscopy and serial section 3D reconstruction. We found contacts between fibroblasts and T2 pneumocytes and between fibroblasts and type 1 pneumocytes that occur at holes in the epithelial basal lamina. The same fibroblast also made contact with pericytes and endothelial cells through similar apertures. A survey of 41 T2 pneumocytes revealed that 54% of T2 pneumocytes had at least one gap in their basal lamina. A morphometric analysis showed these gaps occupied approximately 5.58 +/- 1.51% (mean +/- SE) of the area underneath T2 pneumocytes. We conclude that a population of single fibroblasts link T2 pneumocytes to adjacent capillary endothelial cells in alveolar walls of human lung. We propose that fibroblasts are organized to maintain communication between epithelium and mesenchyme and to provide directional information to migrating leukocytes. PMID- 14551163 TI - Airway inflammation and infection in congenital bilateral absence of the vas deferens. AB - In cystic fibrosis (CF), airway disease begins early in life. Bacteria and elevated levels of neutrophils and inflammatory mediators have been detected in bronchoalveolar lavage (BAL) fluid from infants with CF. Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) are common in men with congenital bilateral absence of the vas deferens (CBAVD) and it has been suggested that this syndrome represents a mild form of CF. We hypothesized that men with CBAVD also have subclinical pulmonary disease. Bronchoscopy with BAL, viral and quantitative bacterial cultures, and analyses of total and differential cell count, cytokines, and free neutrophil elastase was performed in eight men with CBAVD, who had mutations in the CFTR and intermediate or elevated sweat chloride levels, and in four healthy control subjects. There was light growth of Staphylococcus aureus in one of eight men with CBAVD, and small numbers of opportunistic gram-negative bacteria in six of eight men with CBAVD and in one control subject. BAL cell counts and neutrophil elastase were within the normal range. Interleukin-8 and tumor necrosis factor-alpha levels were higher for men with CBAVD than for control subjects. These data suggest that mutations in the CFTR in men with CBAVD, in addition to causing infertility, lead to subclinical bacterial pulmonary infection and inflammation consistent with mild CF. PMID- 14551164 TI - Polymorphism of Clara cell 10-kD protein gene of sarcoidosis. AB - Clara cell 10-kD protein (CC10) exhibits potent antiinflammatory properties. G38A polymorphism was found in the CC10 gene. We investigated the genetic influence of the allele on the development of sarcoidosis using case control analysis in a Japanese population (265 sarcoidosis cases and 258 control subjects). The A allele frequency in sarcoidosis cases (45.1%) was significantly higher than healthy control subjects (34.9%, p = 0.0002). According to outcomes, we divided 223 patients with follow-up periods of 3 years or more into two subgroups (55 progressive and 168 regressive disease). The A allele frequency in patients with progressive disease was significantly higher than control subjects (odds ratio = 4.55; 95% confidence interval, 2.97-6.97; p < 0.0001), whereas that of regressive disease was not. The A/A genotypes had significantly lower bronchoalveolar lavage fluid CC10 levels than the G/G (nonsmokers, p = 0.0054, and smokers, p = 0.0045) and G/A genotypes (nonsmokers, p = 0.0022, and smokers, p = 0.0402). The reporter gene assay showed significantly lower reporter activities in the presence of interferon-gamma for the 38A construct than the 38G construct (p = 0.0177). The G38A polymorphism in the CC10 gene may influence protein expression and be associated with the development of progressive sarcoidosis. PMID- 14551165 TI - Inhaled and nasal corticosteroid use and the risk of fracture. AB - Studies of the risk of fracture associated with inhaled corticosteroids are inconclusive and are limited to short-term effects. We assessed whether long-term use increases this risk. We conducted a case control study nested within a population-based cohort of all Quebec elderly dispensed respiratory medications and followed for at least 4 years during 1988-2001. There were 9,624 new cases of fracture of the hip or upper extremities and 191,622 age-matched control subjects (mean age of 81 years). The rate of any such fracture for current inhaled corticosteroid use was not elevated (rate ratio [RR], 0.97; 95% confidence interval [CI], 0.92-1.03). For upper-extremity fracture, the rate increased by 12% (RR, 1.12; 95% CI, 1.04-1.19) with every 1,000-microg increase in the daily dose of inhaled corticosteroids, but not for hip fracture (RR, 0.97; 95% CI, 0.88 1.07). Among subjects followed for over 8 years, the rate of hip fracture was only elevated with daily doses of more than 2,000 microg of inhaled corticosteroids (RR, 1.61; 95% CI, 1.04-2.50). The rate was not elevated at any dose of nasal corticosteroids. In conclusion, the long-term use of inhaled and nasal corticosteroids at the usual recommended doses is not associated with a risk of fracture in older patients with respiratory disease. PMID- 14551166 TI - Sleep apnea and markers of vascular endothelial function in a large community sample of older adults. AB - Clinical studies have suggested that sleep apnea is associated with impaired brachial artery flow-mediated dilation, a surrogate of endothelial dysfunction. We examined this question among older participants in the baseline examination of the Sleep Heart Health/Cardiovascular Health Study cohort (n = 1,037, age 68 years or older, 56% female). Indices of sleep apnea, derived from 12-channel home polysomnography, were the apnea-hypopnea index (average number of apneas/hypopneas per hour) and the hypoxemia index (percentage of time below 90% O2 saturation). Baseline arterial diameter and percentage of flow-mediated dilation were measured by ultrasound. Sleep apnea measures were associated with baseline diameter and the percentage of flow-mediated dilation, although these associations were weakened after adjustment for other cardiovascular risk factors, particularly body mass index. However, a statistically significant linear association between the hypoxemia index and baseline diameter was observed even after adjustment for body mass index and other confounders (p < 0.01). The associations were stronger among participants who were younger than 80 years and among those who with hypertension. This study adds to the growing body of evidence linking sleep apnea with vascular dysfunction in older subjects. Whether these relationships are entirely independent of obesity is unclear. This association might be one of the mechanisms explaining the relationship between sleep apnea, hypertension, and cardiovascular disease. PMID- 14551167 TI - Endothelial function in obstructive sleep apnea and response to treatment. AB - Impaired endothelium-dependent vascular relaxation is a prognostic marker of atherosclerosis and cardiovascular disease. We evaluated endothelium-dependent flow-mediated dilation (FMD) and endothelium-independent nitroglycerin (NTG) induced dilation of the brachial artery with Doppler ultrasound in 28 men with obstructive sleep apnea (OSA) and 12 men without OSA. Subjects with OSA (apnea hypopnea index; mean +/- SD, 46.0 +/- 14.5) had lower FMD compared with subjects without OSA (5.3 +/- 1.7% vs. 8.3 +/- 1.0%, p < 0.001), and major determinants of FMD were the apnea-hypopnea index and age. There was no significant difference in NTG-induced dilation. Subjects with OSA were randomized to nasal continuous positive airway pressure (nCPAP) or observation for 4 weeks. Subjects on nCPAP had significant increase in FMD, whereas those on observation had no change (4.4% vs. -0.8%, difference of 5.2%, p < 0.001). Neither group showed significant change in NTG-induced vasodilation. Eight subjects who used nCPAP for over 3 months were reassessed on withdrawing treatment for 1 week. On nCPAP withdrawal, FMD became lower than during treatment (p = 0.02) and were similar to baseline values. Our findings demonstrated that men with moderate/severe OSA have endothelial dysfunction and treatment with nCPAP could reverse the dysfunction; the effect, however, was dependent on ongoing use. PMID- 14551168 TI - Exhaled breath condensate detects markers of pulmonary inflammation after cardiothoracic surgery. AB - Cardiac surgery using cardiopulmonary by-pass and, to a greater extent, lung resection, causes acute lung injury that is usually subclinical. Analysis of mediators in exhaled breath condensate is a promising means of monitoring inflammation in a variety of airway diseases but the contribution of the airway lining fluid from the lower respiratory tract is uncertain. We compared the analysis of markers of lung injury in exhaled breath condensate and bronchoalveolar lavage in endotracheally intubated patients before and after coronary artery bypass graft surgery with cardiopulmonary bypass and lobectomy. The neutrophil count and leukotriene B4 concentration in bronchoalveolar lavage fluid rose after coronary artery bypass graft surgery (p < 0.05), but there was no significant change in leukotriene B4, hydrogen peroxide, or hydrogen ion concentrations in exhaled breath condensate. By contrast, after lobectomy, the concentration in exhaled breath condensate of leukotriene B4, hydrogen peroxide and hydrogen ions rose significantly (p < 0.05). Exhaled breath condensate is a safe, noninvasive method of sampling the milieu of the distal lung and is sufficiently sensitive to detect markers of inflammation and oxidative stress in patients after lobectomy, but not after the milder insult associated with cardiac surgery. PMID- 14551169 TI - Effects of moderate exercise and oat beta-glucan on innate immune function and susceptibility to respiratory infection. AB - Both moderate exercise and the soluble oat fiber beta-glucan can increase immune function and decrease risk of infection, but no information exists on their possible combined effects. This study tested the effects of moderate exercise and oat beta-glucan on respiratory infection, macrophage antiviral resistance, and natural killer (NK) cell cytotoxicity. Mice were assigned to four groups: exercise and water, exercise and oat beta-glucan, control water, or control oat beta-glucan. Oat beta-glucan was fed in the drinking water for 10 days before intranasal inoculation of herpes simplex virus type 1 (HSV-1) or euthanasia. Exercise consisted of treadmill running (1 h/day) for 6 days. Macrophage resistance to HSV-1 was increased with both exercise and oat beta-glucan, whereas NK cell cytotoxicity was only increased with exercise. Exercise was also associated with a 45 and 38% decrease in morbidity and mortality, respectively. Mortality was also decreased with oat beta-glucan, but this effect did not reach statistical significance. No additive effects of exercise and oat beta-glucan were found. These data confirm a positive effect of both moderate exercise and oat beta-glucan on immune function, but only moderate exercise was associated with a significant reduction in the risk of upper respiratory tract infection in this model. PMID- 14551170 TI - Stimulation of NPY Y2 receptors by PYY3-36 reveals divergent cardiovascular effects of endogenous NPY in rats on different dietary regimens. AB - In the present experiments the gut hormone peptide YY3-36 (PYY3-36), which inhibits neuropeptide Y (NPY) release, was used as a tool to study the cardiovascular effects of endogenous NPY under different dietary regimens in rats instrumented with a telemetry transmitter. In a first experiment, rats were placed on a standard chow diet ad libitum and in a second experiment on a high fat diet ad libitum. After 6 wk, PYY3-36 (300 microg/kg) or vehicle was injected intraperitoneally. In a third experiment, PYY3-36 or vehicle was administered after 14 days of 50% restriction of a standard chow diet. In food-restricted rats, PYY3-36 increased mean arterial pressure (7 +/- 1 mmHg, mean +/- SE, P < 0.001 vs. saline, 1-way repeated-measures ANOVA with Bonferroni t-test) and heart rate (22 +/- 4 beats/min, P < 0.001) during 3 h after administration. Conversely, PYY3-36 did not influence mean arterial pressure (0 +/- 1 mmHg) and heart rate ( 8 +/- 5 beats/min) significantly in rats on a high-fat diet. Rats fed standard chow diet ad libitum showed an intermediate response (mean arterial pressure 4 +/ 1 mmHg, P < 0.05, and heart rate 5 +/- 2 beats/min, not significant). Thus, in our studies, divergent cardiovascular responses to PYY3-36 were observed in rats on different dietary regimens. These findings suggest that the cardiovascular effects of PYY3-36 depend on the hypothalamic NPY release, which is increased after chronic food restriction and decreased during a high-fat diet. PMID- 14551172 TI - Does the kinin system mediate in cardiovascular abnormalities? An overview. AB - All the components of the kallikrein-kinin system are located in the cardiac muscle, and its deficiency may lead to cardiac dysfunction. In recent years, numerous observations obtained from clinical and experimental models of diabetes, hypertension, cardiac failure, ischemia, myocardial infarction, and left ventricular hypertrophy have suggested that the reduced activity of the local kallikrein-kinin system may be instrumental for the induction of cardiovascular related diseases. The cardioprotective property of the angiotensin-converting enzyme inhibitors is primarily mediated via the kinin-releasing pathway, which may cause regression of left ventricular hypertrophy in hypertensive situations. The ability of kallikrein gene delivery to produce a wide spectrum of beneficial effects makes it an excellent candidate in treating hypertension and cardiovascular and renal diseases. In addition, stable kinin agonists may also be available in the future as therapeutic agents for cardiovascular and renal disorders. PMID- 14551171 TI - Serotonin receptor subtypes required for ventilatory long-term facilitation and its enhancement after chronic intermittent hypoxia in awake rats. AB - Respiratory long-term facilitation (LTF), a serotonin-dependent, persistent augmentation of respiratory activity after episodic hypoxia, is enhanced by pretreatment of chronic intermittent hypoxia (CIH; 5 min 11-12% O2-5 min air, 12 h/night for 7 nights). The present study examined the effects of methysergide (serotonin 5-HT1,2,5,6,7 receptor antagonist), ketanserin (5-HT2 antagonist), or clozapine (5-HT2,6,7 antagonist) on both ventilatory LTF and the CIH effect on ventilatory LTF in conscious male adult rats to determine which specific receptor subtype(s) is involved. In untreated rats (i.e., animals not exposed to CIH), LTF, induced by five episodes of 5-min poikilocapnic hypoxia (10% O2) separated by 5-min normoxic intervals, was measured twice by plethysmography. Thus the measurement was conducted 1-2 days before (as control) and approximately 1 h after systemic injection of methysergide (1 mg/kg ip), ketanserin (1 mg/kg), or clozapine (1.5 mg/kg). Resting ventilation, metabolic rate, and hypoxic ventilatory response (HVR) were unchanged, but LTF ( approximately 18% above baseline) was eliminated by each drug. In CIH-treated rats, LTF was also measured twice, before and approximately 8 h after CIH. Vehicle, methysergide, ketanserin, or clozapine was injected approximately 1 h before the second measurement. Neither resting ventilation nor metabolic rate was changed after CIH and/or any drug. HVR was unchanged after methysergide and ketanserin but reduced in four of seven clozapine rats. The CIH-enhanced LTF ( approximately 28%) was abolished by methysergide and clozapine but only attenuated by ketanserin (to approximately 10%). Collectively, these data suggest that ventilatory LTF requires 5-HT2 receptors and that the CIH effect on LTF requires non-5-HT2 serotonin receptors, probably 5-HT6 and/or 5-HT7 subtype(s). PMID- 14551173 TI - Role of the research-subject locator (RSL) in the performance of a pediatric drug trial. AB - This article examines the feasibility, effectiveness, and ethical constraints relevant to appropriate financial compensation of the pediatric house officer who functions as a research-subject locator (RSL). When carefully trained and supervised, the RSL can facilitate timely enrollment of research subjects in pediatric drug trials, augment the house officer training experience, and advocate for children to increase therapeutic options available for pediatric care. When monitored within the clinical trial schema, the RSL has no direct link to the study outcome and thus performs within ethical standards for subject enrollment and study participation. PMID- 14551174 TI - Can renin status predict the antihypertensive efficacy of eplerenone add-on therapy? AB - Since neither angiotensin-converting enzyme inhibitors (ACE-I) nor angiotensin II receptor blockers (ARB) can completely suppress aldosterone levels, there is a need for alternative/supplementary antihypertensive medications, such as the selective aldosterone blocker eplerenone (Inspra). This multicenter study measured the safety and efficacy of add-on eplerenone therapy to reduce blood pressure not controlled by ACE-I or ARB monotherapy. An ad hoc analysis evaluated whether active plasma renin or serum aldosterone levels could predict blood pressure response to eplerenone therapy. Patients (N = 341) with a diastolic blood pressure > 95 mmHg on a fixed dose of ACE-I or ARB were randomized to 8 weeks of double-blind treatment with eplerenone 50 mg qd or placebo. If blood pressure remained uncontrolled following 2, 4, or 6 weeks of treatment, the eplerenone dose was increased to 100 mg qd. In a combined cohort analysis of these patients, the placebo-adjusted change in systolic and diastolic blood pressure was -5.9/-2.4 mmHg (p< 0.001 and p = 0.006, respectively). While adding eplerenone to an ACE-I or ARB is safe and effective for blood pressure reduction, there was no baseline value or range of values of active plasma renin, serum aldosterone, or their ratio that predicted a favorable response to either of these drug combinations. PMID- 14551175 TI - CYP2C19- and CYP3A4-dependent omeprazole metabolism in West Mexicans. AB - Omeprazole has been used as a drug probe for CYP2C19, but no systematic data are available for Mexican populations. The aim of this study was to evaluate the phenotype frequencies of the CYP2C19 polymorphism in West Mexicans. Besides omeprazole, sulfone was measured to evaluate CYP3A4 after administration of the 20-mg dose to 127 healthy volunteers. Logarithms of metabolic indexes of omeprazole/hydroxyomeprazole for CYP2C19 and omeprazole/omeprazole sulfone for CYP3A4 had trimodal distributions. Five subjects (4%) had a log CYP2C19 metabolic index below -0.9, suggesting an ultra-extensive phenotype. Poor metabolizers (log metabolic index > 0.6) were 6%. For CYP3A4, 11 subjects (9%) were below -0.3 of the log metabolic index. The log metabolic index of omeprazole/omeprazole sulfone was above the antimode of 0.6 for 11% of this population. The mean log metabolic index of CYP3A4 extensive metabolizers (80%) was 0.166, which seems to be higher than the data described for Caucasians and lower than that for Asians. PMID- 14551176 TI - Dose equivalency evaluation of major corticosteroids: pharmacokinetics and cell trafficking and cortisol dynamics. AB - The integrity of current corticosteroid dose equivalency tables, as assessed by mechanistic models for cell trafficking and cortisol dynamics, was investigated in this study. Single, presumably equivalent, doses of intravenous hydrocortisone, methylprednisolone, dexamethasone, and oral prednisolone were given to 5 white men, according to total body weight, in a 5-way crossover, placebo-controlled study. Pharmacodynamic (PD) response-time profiles for T helper cells, T suppressor cells, neutrophils, and adrenal suppression were evaluated by extended indirect response models. For adrenal suppression, prednisolone appears to be less potent than methylprednisolone or dexamethasone. A good correlation was found between the estimated in vivo EC50 values and relative receptor affinity (equilibrium dissociation constants normalized to dexamethasone). Area under the effect curves of all PD responses was calculated using a linear-trapezoidal method. Although T helper cell trafficking and adrenal suppression achieved significant differences by repeated-measures ANOVA (p = 0.014 and 0.022), post hoc analysis using the Bonferroni method revealed no difference between treatments. Although limited by the use of single doses and a relatively small sample size, this study applies mechanistic models for several biomarkers showing that currently used dosing tables reflect reasonable dose equivalency relationships for four corticosteroids. PMID- 14551177 TI - Young and elderly type 2 diabetic patients inhaling insulin with the AERx insulin diabetes management system: a pharmacokinetic and pharmacodynamic comparison. AB - The objective of this study was to compare the pharmacokinetics (PK), pharmacodynamics (PD), and safety of inhaled insulin delivered by the AERx iDMS in young and elderly patients with type 2 diabetes. Twenty-seven young (18-45 years, inclusive) and 28 elderly (>/= 65 years) type 2 diabetic patients were enrolled in this study. A single inhalation of 1.57 mg (45 IU, effect comparable to 6 s.c. units) of regular human insulin was administered to each patient on each of 2 dosing days, and blood samples were drawn up to 360 minutes postdosing to generate the PK/PD curves. AUC(0-360 min) and Cmax values of inhaled insulin were comparable between young and elderly subjects (p = 0.476 for AUC(0-360 min) and p = 0.414 for Cmax). However, the elderly group had significantly less glucose reduction, as indicated by plasma glucose AOC(0-360) (area over the curve) values (p = 0.011). The intrasubject variability of inhaled insulin using the AERx iDMS was similar for young and elderly subjects and was similar to what has previously been reported for soluble insulin administered subcutaneously. Inhaled insulin was well tolerated in these patients, and no changes in pulmonary function tests were observed. A single inhalation of insulin using the AERx iDMS demonstrated comparable insulin PK profiles between the elderly and young type 2 patients but less glucose reduction in the elderly. Based on these results, elderly diabetic patients may need to inhale more insulin than young patients to achieve similar glycemic control. Long-term clinical trials using the AERx device will be useful to study age-related differences. PMID- 14551178 TI - Influence of simulated weightlessness on the oral pharmacokinetics of acetaminophen as a gastric emptying probe in man: a plasma and a saliva study. AB - This study evaluated the effect of simulated weightlessness on gastric emptying, using acetaminophen as a probe and -6 degrees head-down bed rest to simulate zero gravity. Eighteen volunteers were given 1 g of acetaminophen orally before the bed rest and at days 1, 18, and 80. Cmax, tmax, AUC0- infinity, AUC0-t, and t1/2 were calculated for plasma and saliva. The plasma Cmax showed a significant increase (10.43 microg/mL [day 1] to 14.74 microg/mL [day 80]), while tmax significantly decreased (1.41 h [day 1] to 0.91 h [day 80]). Similar results were obtained with saliva, and there were significant increases in the AUCs. The good correlation between the plasma and saliva data suggests that saliva sampling can be valid for acetaminophen pharmacokinetics. The changes in Cmax and tmax indicated more rapid drug absorption, which could have been as a result of faster gastric emptying or an increased blood flow to the intestine. PMID- 14551179 TI - Ragaglitazar: the pharmacokinetics, pharmacodynamics, and tolerability of a novel dual PPAR alpha and gamma agonist in healthy subjects and patients with type 2 diabetes. AB - Ragaglitazar is a novel dual peroxisome proliferator-activated receptor (PPAR) alpha and gamma agonist intended to restore insulin sensitivity and correct diabetic dyslipidemia. These studies assessed single-dose pharmacokinetics and tolerability of ragaglitazar in healthy subjects, as well as multiple-dose pharmacokinetics, pharmacodynamics, and tolerability of ragaglitazar in healthy subjects and in patients with type 2 diabetes. Healthy subjects received a single oral dose (1-120 mg), and healthy subjects and type 2 diabetic patients received a loading dose and thereafter once-daily doses (0.5-16 mg) of ragaglitazar for 6 and 20 days, respectively. Ragaglitazar was rapidly absorbed (tmax: 1.5-1.7 h), with mean AUC0-24 h and Cmax proportional to dose after single and multiple dosing; t1/2 was 80 hours following a single dose and 104 hours in healthy subjects and 122 hours in patients after multiple dosing. Administration of 4 mg ragaglitazar to patients (n = 4) for 21 days resulted in mean decreases from baseline in fasting levels of plasma glucose (18%), C-peptide (18%), fructosamine (6%), triglycerides (36%), free fatty acids (49%), total cholesterol (11%), low density lipoprotein (LDL) cholesterol (21%), and very low-density lipoprotein (VLDL) cholesterol (15%), as well as an increase in high-density lipoprotein (HDL) cholesterol (33%). Overall, ragaglitazar was well tolerated; with multiple dosing, there was a higher incidence of adverse events for patients that, at the highest dose level (16 mg), included peripheral edema and anemia. PMID- 14551180 TI - Plasma kinetics, metabolism, and urinary excretion of alpha-lipoic acid following oral administration in healthy volunteers. AB - R(+)-alpha-lipoic acid is a natural occurring compound that acts as an essential cofactor for certain dehydrogenase complexes. The redox couple alpha-lipoic acid/dihydrolipoic acid possesses potent antioxidant activity. Exogenous racemic alpha-lipoic acid orally administered for the symptomatic treatment of diabetic polyneuropathy is readily and nearly completely absorbed, with a limited absolute bioavailability of about 30% caused by high hepatic extraction. Although the pharmacokinetics of the parent drug have been well characterized in humans, relatively little is known regarding the excretion of alpha-lipoic acid and the pharmacokinetics of any metabolites in humans. In the present study, plasma concentration-time courses, urinary excreted amounts, and pharmacokinetic parameters of alpha-lipoic acid metabolites were evaluated in 9 healthy volunteers after multiple once-daily oral administration of 600 mg racemic alpha lipoic acid. The primary metabolic pathways of alpha-lipoic acid in man, S methylation and beta-oxidation, were quantitatively confirmed by an HPLC electrochemical assay newly established prior to the beginning of this study. Major circulating metabolites were the S-methylated beta-oxidation products 4,6 bismethylthio-hexanoic acid and 2,4-bismethylthio-butanoic acid, whereas its conjugated forms accounted for the major portion excreted in urine. There was no statistically significant difference in the pharmacokinetic parameters Cmax, AUC, and tmax between day 1 and day 4. Despite the prolonged half-lives of the major metabolites compared to the parent drug, no evidence of accumulation was found. Mean values of 12.4% of the administered dose were recovered in the urine after 24 hours as the sum of alpha-lipoic acid and its metabolites. The results of the present study revealed that urinary excretion of alpha-lipoic acid and five of its main metabolites does not play a significant role in the elimination of alpha lipoic acid. Therefore, biliary excretion, further electrochemically inactive degradation products, and complete utilization of alpha-lipoic acid as a primary substrate in the endogenous metabolism should be considered. PMID- 14551181 TI - Individual difference in the pharmacokinetics of a drug, pravastatin, in healthy subjects. AB - The purpose of this investigation was to determine whether there were individual pharmacokinetic differences of a drug, pravastatin. Furthermore, the percentage of subjects who showed pharmacokinetic differences was determined. A single oral dose of pravastatin 10 mg was administered to 84 Japanese healthy male subjects. Serum concentrations of pravastatin were measured for 8 hours postdose. Area under the concentration-time curve (AUC) and peak concentration (Cmax) were determined as primary evaluation parameters. An outlier was defined as follows: Outlier 1 < Q1 - (Q3 - Q1) x 1.5 or Q3+(Q3-Q1) x 1.5 < Outlier 1, Outlier 2 < Q1 (Q3 - Q1) x 3 or Q3+ (Q3-Q1) x 3 < Outlier 2. Subjects who were outliers were regarded as having an individual difference in pharmacokinetic behavior. In AUC and Cmax, 4 of 84 subjects (4.8%) were higher outliers. Of these 4 subjects, 2 were high outliers in both AUC and Cmax. No subjects were low outliers. It was concluded that a significant individual difference in the pharmacokinetics of pravastatin was observed in 4.8% of the subjects (4/84). PMID- 14551182 TI - Pharmacokinetic and pharmacodynamic interactions of oral midazolam with ketoconazole, fluoxetine, fluvoxamine, and nefazodone. AB - The objective of this study was to investigate pharmacokinetic and pharmacodynamic interactions between midazolam and fluoxetine, fluvoxamine, nefazodone, and ketoconazole. Forty healthy subjects were randomized to receive one of the four study drugs for 12 days in a parallel study design: fluoxetine 60 mg per day for 5 days, followed by 20 mg per day for 7 days; fluvoxamine titrated to a daily dose of 200 mg; nefazodone titrated to a daily dose of 400 mg; or ketoconazole 200 mg per day. All 40 subjects received oral midazolam solution before and after the 12-day study drug regimen. Blood samples for determination of midazolam concentrations were drawn for 24 hours after each midazolam dose and used for the calculation of pharmacokinetic parameters. The effects of the study drugs on midazolam pharmacodynamics were assessed using the symbol digit modalities test (SDMT). The mean area under the curve (AUC) for midazolam was increased 771.9% by ketoconazole and 444.0% by nefazodone administration. However, there was no significant change in midazolam AUC as a result of fluoxetine (13.4% decrease) and a statistical trend for fluvoxamine (66.1% increase) administration. Pharmacodynamic data are consistent with pharmacokinetic data indicating that nefazodone and ketoconazole resulted in significant increases in midazolam-related cognition impairment. The significant impairment in subjects' cognitive function reflects the changes in midazolam clearance after treatment with ketoconazole and nefazodone. These results suggest that caution with the use of midazolam is warranted with potent CYP3A4 inhibitors. PMID- 14551183 TI - Influence of goldenseal root on the pharmacokinetics of indinavir. AB - Goldenseal root was identified as the most potent inhibitor of CYP3A4 in a study that tested 21 popular herbal products for in vitro inhibitory activity. The purpose of this investigation was to examine the influence of goldenseal root on the disposition of the CYP3A4 substrate indinavir in humans. Using a crossover study design, the pharmacokinetics of indinavir were characterized in 10 healthy volunteers before and after 14 days of treatment with goldenseal root (1140 mg twice daily). Indinavir was given as a single 800-mg oral dose, and blood samples were collected for 8 hours following the dose. No statistically significant differences in peak concentration (11.6 vs. 11.9 mg/L) or oral clearance (26.8 vs. 23.9 mg*h/L) were observed following treatment with goldenseal root. Half life and time to reach peak concentration were also unchanged by goldenseal. These results suggest that patients being treated with indinavir can safely take goldenseal root and that interactions with other drugs metabolized by CYP3A4 in the liver are unlikely. PMID- 14551184 TI - Identification of tyrosine sulfation in extracellular leucine-rich repeat proteins using mass spectrometry. AB - Multiple and variable tyrosine sulfation in extracellular class II leucine-rich repeat proteins/proteoglycans were characterized by mass spectrometry. The sulfogroup on tyrosine is labile and is released from peptides under normal mass spectrometric conditions. Thus, special approaches must be considered in order to identify this modification. By using a combination of mass spectrometry studies operating in negative and positive ion mode, tyrosine sulfation could be identified. In positive mode, the peptides normally appeared non-sulfated, whereas in negative mode a mixture of sulfated and non-sulfated species was observed. A combination of peptides released by different proteinases was used to obtain details on the locations of sulfate groups. Multiple tyrosine sulfates were observed in the N-terminal region of fibromodulin (up to 9 sites), osteoadherin (up to 6 sites), and lumican (2 sites). Osteoadherin contains two additional sulfated tyrosine residues close to its C terminus. We also identified an error in the published sequence of bovine fibromodulin, resulting in the replacement of Thr37 by Tyr37-Gly38, thus increasing its homology with its human counterpart. PMID- 14551185 TI - Initiation of mucin-type O-glycosylation in dictyostelium is homologous to the corresponding step in animals and is important for spore coat function. AB - Like animal cells, many unicellular eukaryotes modify mucin-like domains of secretory proteins with multiple O-linked glycans. Unlike animal mucin-type glycans, those of some microbial eukaryotes are initiated by alpha-linked GlcNAc rather than alpha-GalNAc. Based on sequence similarity to a recently cloned soluble polypeptide hydroxyproline GlcNAc-transferase that modifies Skp1 in the cytoplasm of the social ameba Dictyostelium, we have identified an enzyme, polypeptide alpha-N-acetylglucosaminyltransferase (pp alpha-GlcNAc-T2), that attaches GlcNAc to numerous secretory proteins in this organism. Unlike the Skp1 GlcNAc-transferase, pp alpha-GlcNAc-T2 is predicted to be a type 2 transmembrane protein. A highly purified, soluble, recombinant fragment of pp alpha-GlcNAc-T2 efficiently transfers GlcNAc from UDP-GlcNAc to synthetic peptides corresponding to mucin-like domains in two proteins that traverse the secretory pathway. pp alpha-GlcNAc-T2 is required for addition of GlcNAc to peptides in cell extracts and to the proteins in vivo. Mass spectrometry and Edman degradation analyses show that pp alpha-GlcNAc-T2 attaches GlcNAc in alpha-linkage to the Thr residues of all the synthetic mucin repeats. pp alpha-GlcNAc-T2 is encoded by the previously described modB locus defined by chemical mutagenesis, based on sequence analysis and complementation studies. This finding establishes that the many phenotypes of modB mutants, including a permeability defect in the spore coat, can now be ascribed to defects in mucin-type O-glycosylation. A comparison of the sequences of pp alpha-GlcNAc-T2 and the animal pp alpha-GalNAc transferases reveals an ancient common ancestry indicating that, despite the different N-acetylhexosamines involved, the enzymes share a common mechanism of action. PMID- 14551186 TI - Direct measurement of antigen binding properties of CD1 proteins using fluorescent lipid probes. AB - CD1 proteins are antigen-presenting molecules that bind foreign and self-lipids and stimulate specific T cell responses. In the current study, we investigated ligand binding by CD1 proteins by developing a fluorescent probe binding approach using soluble recombinant human CD1 proteins. To increase stability and yield, soluble group 1 CD1 (CD1b and CD1c) and group 2 CD1 (CD1d) proteins were produced as single chain secreted CD1 proteins in which beta2-microglobulin was fused to the N termini of the CD1 heavy chains by a flexible peptide linker sequence. Analysis of ligand binding properties of single chain secreted CD1 proteins by using fluorescent lipid probes indicated significant differences in ligand preference and in pH dependence of binding by group 1 versus group 2 CD1 proteins. Whereas group 1 CD1 isoforms (CD1b and CD1c) show stronger binding of nitrobenzoxadiazole (NBD)-labeled dialkyl-based ligands (phosphatidylcholine, sphingomyelin, and ceramide), group 2 CD1 (CD1d) proteins were stronger binders of small hydrophobic probes such as 1-anilinonaphthalene-8-sulfonic acid and 4,4' dianilino-1,1'-naphthyl-5,5'-disulfonic acid. Competition studies indicated that binding of fluorescent lipid probes involved association of the probe with the hydrophobic ligand binding groove of CD1 proteins. Analysis of selected alanine substitution mutants of human CD1b known to inhibit antigen presentation showed that NBD-labeled lipid probe binding could be used to distinguish mutations that interfere with ligand binding from those that affect T cell receptor docking. Our findings provide further evidence for the functional specialization of different CD1 isoforms and demonstrate the value of the fluorescent lipid probe binding method for assisting structure-based studies of CD1 function. PMID- 14551187 TI - Mechanistic basis for reduced viral and enzymatic fitness of HIV-1 reverse transcriptase containing both K65R and M184V mutations. AB - HIV-1 drug resistance mutations are often inversely correlated with viral fitness, which remains poorly described at the molecular level. Some resistance mutations can also suppress resistance caused by other resistance mutations. We report the molecular mechanisms by which a virus resistant to lamivudine with the M184V reverse transcriptase mutation shows increased susceptibility to tenofovir and can suppress the effects of the tenofovir resistance mutation K65R. Additionally, we report how the decreased viral replication capacity of resistant viruses is directly linked to their decreased ability to use natural nucleotide substrates and that combination of the K65R and M184V resistance mutations leads to greater decreases in viral replication capacity. All together, these results define at the molecular level how nucleoside-resistant viruses can be driven to reduced viral fitness. PMID- 14551188 TI - Expression and purification of enzymatically active forms of the human lysyl oxidase-like protein 4. AB - The lysyl oxidase-like protein 4 (LOXL4) is the latest member of the emerging family of lysyl oxidases, several of which were shown to function as copper dependent amine oxidases catalyzing lysine-derived cross-links in extracellular matrix proteins. LOXL4 contains four scavenger receptor cysteine-rich domains in addition to the characteristic domains of the LOX family, including the copper binding domain, the cytokine receptor-like domain, and the residues of the lysyl tyrosyl quinone cofactor. In an effort to assess its amine oxidase activity, we expressed LOXL4 as recombinant forms attached with hexa-histidine residues at the carboxyl terminus by using an Escherichia coli expression system. The recombinant proteins were purified with nickel-chelating affinity chromatography and converted into enzymatically active forms by stepwise dialysis. The purified LOXL4 proteins showed beta-aminopropionitrile-inhibitable activity of 0.022-0.032 units/mg toward a nonpeptidyl substrate, benzylamine. These results indicate that LOXL4, with the four scavenger receptor cysteine rich domains, may also function as an active amine oxidase. Availability of the pure and active forms of LOXL4 will be significantly helpful in functional studies related to substrate specificity and crystal structure of this amine oxidase, which should provide significant insights into functional differences within the LOX family members. PMID- 14551189 TI - Genetic locus and structural characterization of the biochemical defect in the O antigenic polysaccharide of the symbiotically deficient Rhizobium etli mutant, CE166. Replacement of N-acetylquinovosamine with its hexosyl-4-ulose precursor. AB - The O-antigen polysaccharide (OPS) of Rhizobium etli CE3 lipopolysaccharide (LPS) is linked to the core oligosaccharide via an N-acetylquinovosaminosyl (QuiNAc) residue. A mutant of CE3, CE166, produces LPS with reduced amounts of OPS, and a suppressed mutant, CE166 alpha, produces LPS with nearly normal OPS levels. Both mutants are deficient in QuiNAc production. Characterization of OPS from CE166 and CE166 alpha showed that QuiNAc was replaced by its 4-keto derivative, 2 acetamido-2,6-dideoxyhexosyl-4-ulose. The identity of this residue was determined by NMR and mass spectrometry, and by gas chromatography-mass spectrometry analysis of its 2-acetamido-4-deutero-2,6-dideoxyhexosyl derivatives produced by reduction of the 4-keto group using borodeuteride. Mass spectrometric and methylation analyses showed that the 2-acetamido-2,6-dideoxyhexosyl-4-ulosyl residue was 3-linked and attached to the core-region external Kdo III residue of the LPS, the same position as that of QuiNAc in the CE3 LPS. DNA sequencing revealed that the transposon insertion in strain CE166 was located in an open reading frame whose predicted translation product, LpsQ, falls within a large family of predicted open reading frames, which includes biochemically characterized members that are sugar epimerases and/or reductases. A hypothesis to be tested in future work is that lpsQ encodes UDP-2-acetamido-2,6 dideoxyhexosyl-4-ulose reductase, the second step in the synthesis of UDP-QuiNAc from UDP-GlcNAc. PMID- 14551190 TI - Reduced folate carrier gene silencing in multiple antifolate-resistant tumor cell lines is due to a simultaneous loss of function of multiple transcription factors but not promoter methylation. AB - The human reduced folate carrier (hRFC) is the major uptake route for antifolates used in cancer chemotherapy. Here we explored the molecular basis for the decrease or loss of hRFC gene expression in seventeen tumor cell lines with resistance to multiple antifolates due to impaired antifolate transport. We studied the role of various cis-acting elements including CRE/AP-1-like element and GC-box in hRFC promoters A and B, respectively, as well as AP-2, Mzf-1 and E box that are contained within or near four tandemly repeated sequences upstream of promoter A. Decreased or abolished binding either to [32P]GC-box, Mzf-1, AP-1, E-box, or CRE oligonucleotides was detected in approximately 50-80% of antifolate resistant cell lines. Strikingly, approximately 80% of the cell lines displayed a simultaneously decreased binding to three or more of these hRFC promoter elements, whereas normal AP-2 binding was retained. The possible contribution of promoter methylation to hRFC gene silencing was also explored. None of the antifolate-resistant cell lines, except for MDA-MB-231 cells, showed hRFC promoter methylation; consistently, MDA-MB-231 was the only cell line that retained binding to all six cis-acting elements. Western blot analysis demonstrated decreased expression of transcriptional activators (pCREB-1, pATF-1, USF-1, c-Fos, c-Jun, Sp1, and Sp3) and/or increased expression of repressors (short Sp3 isoforms), whereas normal AP2alpha levels were retained. Transient expression of the relevant transcription factors restored, at least partially, both promoter binding and hRFC gene expression. This is the first report that transcriptional silencing of the hRFC gene in multiple tumor cell lines with resistance to various novel antifolates is a result of a simultaneous loss of function of multiple transcription factors but not promoter methylation. PMID- 14551191 TI - PINCH-1 is an obligate partner of integrin-linked kinase (ILK) functioning in cell shape modulation, motility, and survival. AB - PINCH-1 is a widely expressed focal adhesion protein that forms a ternary complex with integrin-linked kinase (ILK) and CH-ILKBP/actopaxin/alpha-parvin (abbreviated as alpha-parvin herein). We have used RNA interference, a powerful approach of reverse genetics, to investigate the functions of PINCH-1 and ILK in human cells. We report here the following. First, PINCH-1 and ILK, but not alpha parvin, are essential for prompt cell spreading and motility. Second, PINCH-1 and ILK, like alpha-parvin, are crucial for cell survival. Third, PINCH-1 and ILK are required for optimal activating phosphorylation of PKB/Akt, an important signaling intermediate of the survival pathway. Whereas depletion of ILK reduced Ser473 phosphorylation but not Thr308 phosphorylation of PKB/Akt, depletion of PINCH-1 reduced both the Ser473 and Thr308 phosphorylation of PKB/Akt. Fourth, PINCH-1 and ILK function in the survival pathway not only upstream but also downstream (or in parallel) of protein kinase B (PKB)/Akt. Fifth, PINCH-1, ILK and to a less extent alpha-parvin are mutually dependent in maintenance of their protein, but not mRNA, levels. The coordinated down-regulation of PINCH-1, ILK, and alpha-parvin proteins is mediated at least in part by proteasomes. Finally, increased expression of PINCH-2, an ILK-binding protein that is structurally related to PINCH-1, prevented the down-regulation of ILK and alpha-parvin induced by the loss of PINCH-1 but failed to restore the survival signaling or cell shape modulation. These results provide new insights into the functions of PINCH proteins in regulation of ILK and alpha-parvin and control of cell behavior. PMID- 14551192 TI - Epidermal growth factor induction of phenotype-dependent cell cycle arrest in vascular smooth muscle cells is through the mitogen-activated protein kinase pathway. AB - The heterogeneity of vascular smooth muscle cells is well established in tissue culture, but their differential responses to growth factors are not completely defined. We wished to identify effects of epidermal growth factor (EGF) on vascular smooth muscle cells in distinct phenotypes, such as spindle and epithelioid. We found that the EGF receptors were abundant in epithelioid cells but not spindle cells. EGF treatment inhibited serum-independent DNA synthesis, which was absent in spindle cells, of epithelioid cells. Additionally, using a pulse-chase assay, we found that bromodeoxyuridine-labeled cells failed to re enter the S phase in the presence of EGF. These EGF effects were abolished by either inhibiting the EGF receptor tyrosine kinase with AG1478 or inhibiting the mitogen-activated protein kinase pathway with PD98059. In response to treatment with EGF, the EGF receptor was phosphorylated, which was correlated with phosphorylation and activation of p42/44 mitogen-activated protein kinases. Inhibition of EGF receptor phosphorylation and mitogen-activated protein kinase activation resulted in a reversal of the EGF-induced inhibition of bromodeoxyuridine incorporation and cell cycle arrest. Subsequent studies revealed that the activation of the EGF receptor and the mitogen-activated protein kinase pathway in epithelioid cells induced expression of the cell cycle inhibitory protein p27Kip1 but not p21Cip1. Taken together, our data demonstrate that the EGF receptor is abundantly expressed in epithelioid vascular smooth muscle cells and that the activation of this receptor results in cell cycle arrest through activation of the mitogen-activated protein kinase pathway. PMID- 14551193 TI - A Bax/Bak-independent mitochondrial death pathway triggered by Drosophila Grim GH3 domain in mammalian cells. AB - Grim encodes a protein required for programmed cell death in Drosophila, whose proapoptotic activity is conserved in mammalian cells. Two proapoptotic domains are relevant for Grim killing function; the N-terminal region, which induces apoptosis by disrupting inhibitor of apoptosis protein (IAP) blockage of caspase activity, and the internal GH3 domain, which triggers a mitochondrial pathway. We explored the role of these two domains in heterologous killing of mammalian cells by Grim. The GH3 domain is essential for Grim proapoptotic activity in mouse cells, whereas the N-terminal domain is dispensable. The GH3 domain is required and sufficient for Grim targeting to mitochondria and for cytochrome c release in a caspase- and N-terminal-independent, IAP-insensitive manner. These Grim GH3 activities do not require Bax or Bak function, revealing GH3 activity as the first proapoptotic stimulus able to trigger the mitochondrial death pathway in mammalian cells in the absence of multidomain proapoptotic Bcl-2 proteins. PMID- 14551194 TI - Bifunctional phosphoglucose/phosphomannose isomerases from the Archaea Aeropyrum pernix and Thermoplasma acidophilum constitute a novel enzyme family within the phosphoglucose isomerase superfamily. AB - The hyperthermophilic crenarchaeon Aeropyrum pernix contains phosphoglucose isomerase (PGI) activity. However, obvious homologs with significant identity to known PGIs could not be identified in the sequenced genome of this organism. The PGI activity from A. pernix was purified and characterized. Kinetic analysis revealed that, unlike all known PGIs, the enzyme catalyzed reversible isomerization not only of glucose 6-phosphate but also of epimeric mannose 6 phosphate at similar catalytic efficiency, thus defining the protein as bifunctional phosphoglucose/phosphomannose isomerase (PGI/PMI). The gene pgi/pmi encoding PGI/PMI (open reading frame APE0768) was identified by matrix-assisted laser desorption ionization time-of-flight analyses; the gene was overexpressed in Escherichia coli as functional PGI/PMI. Putative PGI/PMI homologs were identified in several (hyper)thermophilic archaea and two bacteria. The homolog from Thermoplasma acidophilum (Ta1419) was overexpressed in E. coli, and the recombinant enzyme was characterized as bifunctional PGI/PMI. PGI/PMIs showed low sequence identity to the PGI superfamily and formed a distinct phylogenetic cluster. However, secondary structure predictions and the presence of several conserved amino acids potentially involved in catalysis indicate some structural and functional similarity to the PGI superfamily. Thus, we propose that bifunctional PGI/PMI constitutes a novel protein family within the PGI superfamily. PMID- 14551195 TI - Targeted disruption of the PDZK1 gene in mice causes tissue-specific depletion of the high density lipoprotein receptor scavenger receptor class B type I and altered lipoprotein metabolism. AB - PDZK1, a multi-PDZ domain containing adaptor protein, interacts with various membrane proteins, including the high density lipoprotein (HDL) receptor scavenger receptor class B type I (SR-BI). Here we show that PDZK1 controls in a tissue-specific and post-transcriptional fashion the expression of SR-BI in vivo. SR-BI protein expression in PDZK1 knock-out (KO) mice was reduced by 95% in the liver, 50% in the proximal intestine, and not affected in steroidogenic organs (adrenal, ovary, and testis). Thus, PDZK1 joins a growing list of adaptors that control tissue-specific activity of cell surface receptors. Hepatic expression of SR-BII, a minor splice variant with an alternative C-terminal cytoplasmic domain, was not affected in PDZK1 KO mice, suggesting that binding of PDZK1 to SR-BI is required for controlling hepatic SR-BI expression. The loss of hepatic SR-BI was the likely cause of the elevation in plasma total and HDL cholesterol and the increase in HDL particle size in PDZK1 KO mice, phenotypes similar to those observed in SR-BI KO mice. PDZK1 KO mice differed from SR-BI KO mice in that the ratio of unesterified to total plasma cholesterol was normal, females were fertile, and cholesteryl ester stores in steroidogenic organs were essentially unaffected. These differences may be due to nearly normal extrahepatic expression of SR-BI in PDZK1 KO mice. The PDZK1-dependent regulation of hepatic SR-BI and, thus, lipoprotein metabolism supports the proposal that this adaptor may represent a new target for therapeutic intervention in cardiovascular disease. PMID- 14551196 TI - Disruption of aldehyde reductase increases group size in dictyostelium. AB - Developing Dictyostelium cells form structures containing approximately 20,000 cells. The size regulation mechanism involves a secreted counting factor (CF) repressing cytosolic glucose levels. Glucose or a glucose metabolite affects cell cell adhesion and motility; these in turn affect whether a group stays together, loses cells, or even breaks up. NADPH-coupled aldehyde reductase reduces a wide variety of aldehydes to the corresponding alcohols, including converting glucose to sorbitol. The levels of this enzyme previously appeared to be regulated by CF. We find that disrupting alrA, the gene encoding aldehyde reductase, results in the loss of alrA mRNA and AlrA protein and a decrease in the ability of cell lysates to reduce both glyceraldehyde and glucose in an NADPH-coupled reaction. Counterintuitively, alrA- cells grow normally and have decreased glucose levels compared with parental cells. The alrA- cells form long unbroken streams and huge groups. Expression of AlrA in alrA- cells causes cells to form normal fruiting bodies, indicating that AlrA affects group size. alrA- cells have normal adhesion but a reduced motility, and computer simulations suggest that this could indeed result in the formation of large groups. alrA- cells secrete low levels of countin and CF50, two components of CF, and this could partially account for why alrA- cells form large groups. alrA- cells are responsive to CF and are partially responsive to recombinant countin and CF50, suggesting that disrupting alrA inhibits but does not completely block the CF signal transduction pathway. Gas chromatography/mass spectroscopy indicates that the concentrations of several metabolites are altered in alrA- cells, suggesting that the Dictyostelium aldehyde reductase affects several metabolic pathways in addition to converting glucose to sorbitol. Together, our data suggest that disrupting alrA affects CF secretion, causes many effects on cellular metabolism, and has a major effect on group size. PMID- 14551197 TI - Physical and functional interaction between Hck tyrosine kinase and guanine nucleotide exchange factor C3G results in apoptosis, which is independent of C3G catalytic domain. AB - The hematopoietic cell kinase Hck is a Src family tyrosine kinase expressed in cells of myelomonocytic lineage, B lymphocytes, and embryonic stem cells. To study its role in signaling pathways we used the Hck-SH3 domain in protein interaction cloning and identified C3G, the guanine nucleotide exchange factor for Rap1 and R-Ras, as a protein that associated with Hck. This interaction was direct and was mediated partly through the proline-rich region of C3G. C3G could be co-immunoprecipitated with Hck from Cos-1 cells transfected with Hck and C3G. C3G was phosphorylated on tyrosine 504 in cells when coexpressed with Hck but not with a catalytically inactive mutant of Hck. Phosphorylation of endogenous C3G at Tyr-504 was increased by treatment of human myelomonocytic THP-1 cells with mercuric chloride, which is known to activate Hck tyrosine kinase specifically. Coexpression of Hck with C3G induced a high level of apoptosis in many cell lines by 30-42 h of transfection. Induction of apoptosis was not dependent on Tyr-504 phosphorylation or the catalytic domain of C3G but required the catalytic activity of Hck. Using dominant negative constructs of caspases we found that caspase-1, -8, and -9 are involved in this apoptotic pathway. These results suggest that C3G and Hck interact physically and functionally in vivo to activate kinase-dependent and caspase-mediated apoptosis, which is independent of catalytic domain of C3G. PMID- 14551198 TI - Lipid peroxidation during the hypersensitive response in potato in the absence of 9-lipoxygenases. AB - Hypersensitive cell death is an important defense reaction of plants to pathogen infection and is accompanied by lipid peroxidation processes. These may occur non enzymatically by the action of reactive oxygen species or may be catalyzed by enzymes such as alpha-dioxygenases, lipoxygenases, or peroxidases. Correlative data showing increases in 9-lipoxygenase products in hyper-sensitively reacting cells have so far suggested that a large part of lipid peroxidation is mediated by a specific set of 9-lipoxygenases. To address the significance of 9 lipoxygenases for this type of pathogen response in potato, RNA interference constructs of a specific pathogen-induced potato 9-lipoxygenase were transferred to potato plants. Significantly reduced 9-lipoxygenase transcript levels were observed in transgenic plants after pathogen treatment. In addition, 9 lipoxygenase activity was hardly detectable, and levels of 9-lipoxygenase-derived oxylipins were reduced up to 12-fold after pathogen infection. In contrast to wild type plants, high levels of non-enzymatically as well as 13-lipoxygenase derived oxylipins were present in 9-lipoxygenase-deficient plants. From this we conclude that during the normal hypersensitive response in potato, lipid peroxidation may occur as a controlled and directed process that is facilitated by the action of a specific 9-lipoxygenase. If 9-lipoxygenase-mediated formation of hydroperoxides is repressed, autoxidative lipid peroxidation processes and 13 lipoxygenase-mediated oxylipins synthesis become prominent. The unaltered timing and extent of necrosis formation suggests that the origin of lipid hydroperoxides does not influence pathogen-induced cell death in potato. PMID- 14551199 TI - High affinity interaction of syntaxin and SNAP-25 on the plasma membrane is abolished by botulinum toxin E. AB - The release of hormones and neurotransmitters requires the fusion of cargo containing vesicles with the plasma membrane. This process of exocytosis relies on three SNARE proteins, namely syntaxin and SNAP-25 on the target plasma membrane and synaptobrevin on the vesicular membrane. In this study we examined the molecular assembly pathway that leads to formation of the fusogenic SNARE complex. We now show that the plasma membrane syntaxin and SNAP-25 interact with high affinity and equimolar stoichiometry to form a stable dimer on the pathway to the ternary SNARE complex. In bovine chromaffin cells, syntaxin and SNAP-25 colocalize in defined clusters that average 700 nm in diameter and cover 10% of the plasma membrane. Removal of the C terminus of SNAP-25 by botulinum neurotoxin E, a known neuroparalytic agent, dissociates the target SNARE dimer in vitro and disrupts the SNARE clustering in vivo. Together, our data uncover formation of stable syntaxin/SNAP-25 dimers as a central principle of the SNARE assembly pathway underlying regulated exocytosis. PMID- 14551201 TI - Role for Fes/Fps tyrosine kinase in microtubule nucleation through is Fes/CIP4 homology domain. AB - We have previously demonstrated that Fes/Fps (Fes) tyrosine kinase is involved in Semaphorin3A-mediated signaling. Here we report a role for Fes tyrosine kinase in microtubule dynamics. A fibrous formation of Fes was observed in a kinase dependent manner, which associated with microtubules and functionally correlated with microtubule bundling. Microtubule regeneration assays revealed that Fes aggregates colocalized with gamma-tubulin at microtubule nucleation sites in a Fes/CIP4 homology (FCH) domain-dependent manner and that expression of FCH domain deleted Fes mutants blocked normal centrosome formation. In support of these observations, mouse embryonic fibroblasts derived from Fes-deficient mice displayed an aberrant structure of nucleation and centrosome with unbundling and disoriented filaments of microtubules. Our findings suggest that Fes plays a critical role in microtubule dynamics including microtubule nucleation and bundling through its FCH domain. PMID- 14551200 TI - Differential involvement of the Ras and Rap1 small GTPases in vasoactive intestinal and pituitary adenylyl cyclase activating polypeptides control of the prolactin gene. AB - In pituitary cells, transcriptional regulation of the prolactin (PRL) gene and prolactin secretion are controlled by multiple transduction pathways through the activation of G protein coupled receptors and receptor tyrosine kinases. In the somatolactotrope GH4C1 cell line, we have previously identified crosstalk between the MAPKinase cascade ERK1/2 and the cAMP/protein kinase A pathway after the activation of the VPAC2 receptor by vasoactive intestinal polypeptide (VIP) or pituitary adenylyl cyclase-activating polypeptide (PACAP38). In the present study, we focus on the involvement of the GTPases Ras and Rap1 as downstream components of signal transmission initiated by activation of the VPAC2 receptor. By using pull-down experiments, we show that VIP and PACAP38 preferentially activate Rap1, whereas thyrotropin releasing hormone (TRH) and epidermal growth factor (EGF) mainly activate Ras GTPase. Experiments involving the expression of the dominant-negative mutants of Ras and Rap1 signaling (RasN17 or Rap1N17) indicate that both GTPases Ras and Rap1 are recruited for the ERK activation by VIP and PACAP38, whereas Rap1 is poorly involved in TRH or EGF-induced ERK activation. The use of U0126, a selective inhibitor of MAPKinase kinase, provides evidence that MAPKinase contributes to the regulation of the PRL gene. Moreover, cotransfection of RasN17 or Rap1N17 with the PRL proximal promoter luciferase reporter construct indicates that Rap1 may be responsible for VIP/PACAP-induced activation of the PRL promoter. Interestingly, Ras would be involved as a negative regulator of VIP/PACAP-induced PRL gene activation, in contrast to its stimulatory role in the regulation of the PRL promoter by TRH and EGF. PMID- 14551202 TI - A new role for IQ motif proteins in regulating calmodulin function. AB - IQ motifs are found in diverse families of calmodulin (CaM)-binding proteins. Some of these, like PEP-19 and RC3, are highly abundant in neuronal tissues, but being devoid of catalytic activity, their biological roles are not understood. We hypothesized that these IQ motif proteins might have unique effects on the Ca2+ binding properties of CaM, since they bind to CaM in the presence or absence of Ca2+. Here we show that PEP-19 accelerates by 40 to 50-fold both the slow association and dissociation of Ca2+ from the C-domain of free CaM, and we identify the sites of interaction between CaM and PEP-19 using NMR. Importantly, we demonstrate that PEP-19 can also increase the rate of dissociation of Ca2+ from CaM when bound to intact CaM-dependent protein kinase II. Thus, PEP-19, and presumably similar members of the IQ family of proteins, has the potential to alter the Ca2+-binding dynamics of free CaM and CaM that is bound to other target proteins. Since Ca2+ binding to the C-domain of CaM is the rate-limiting step for activation of CaM-dependent enzymes, the data reveal a new concept of importance in understanding the temporal dynamics of Ca2+-dependent cell signaling. PMID- 14551203 TI - Inactivation of NADP+-dependent isocitrate dehydrogenase by peroxynitrite. Implications for cytotoxicity and alcohol-induced liver injury. AB - Recently, we demonstrated that the control of cytosolic and mitochondrial redox balance and oxidative damage is one of the primary functions of NADP+-dependent isocitrate dehydrogenase (ICDH) by supplying NADPH for antioxidant systems. We investigated whether the ICDH would be a vulnerable target of peroxynitrite anion (ONOO-) as a purified enzyme, in intact cells, and in liver mitochondria from ethanol-fed rats. Synthetic peroxynitrite and 3-morpholinosydnomine N ethylcarbamide (SIN-1), a peroxynitrite-generating compound, inactivated ICDH in a dose- and time-dependent manner. The inactivation of ICDH by peroxynitrite or SIN-1 was reversed by dithiothreitol. Loss of enzyme activity was associated with the depletion of the thiol groups in protein. Immunoblotting analysis of peroxynitrite-modified ICDH indicates that S-nitrosylation of cysteine and nitration of tyrosine residues are the predominant modifications. Using electrospray ionization mass spectrometry (ESI-MS) with tryptic digestion of protein, we found that peroxynitrite forms S-nitrosothiol adducts on Cys305 and Cys387 of ICDH. Nitration of Tyr280 was also identified, however, this modification did not significantly affect the activity of ICDH. These results indicate that S-nitrosylation of cysteine residues on ICDH is a mechanism involving the inactivation of ICDH by peroxynitrite. The structural alterations of modified enzyme were indicated by the changes in protease susceptibility and binding of the hydrophobic probe 8-anilino-1-napthalene sulfonic acid. When U937 cells were incubated with 100 microM SIN-1 bolus, a significant decrease in both cytosolic and mitochondrial ICDH activities were observed. Using immunoprecipitation and ESI-MS, we were also able to isolate and positively identify S-nitrosylated and nitrated mitochondrial ICDH from SIN-1-treated U937 cells as well as liver from ethanol-fed rats. Inactivation of ICDH resulted in the pro-oxidant state of cells reflected by an increased level of intracellular reactive oxygen species, a decrease in the ratio of [NADPH]/[NADPH + NADP+], and a decrease in the efficiency of reduced glutathione turnover. The peroxynitrite mediated damage to ICDH may result in the perturbation of the cellular antioxidant defense mechanisms and subsequently lead to a pro-oxidant condition. PMID- 14551204 TI - Jak2 tyrosine kinase mediates angiotensin II-dependent inactivation of ERK2 via induction of mitogen-activated protein kinase phosphatase 1. AB - Previous work has shown that inhibition of Jak2 via the pharmacological compound AG490 blocks the angiotensin II (Ang II)-dependent activation of ERK2, thereby suggesting an essential role of Jak2 in ERK activation. However, recent studies have thrown into question the specificity of AG490 and therefore the role of Jak2 in ERK activation. To address this, we reconstituted an Ang II signaling system in a Jak2-/-cell line and measured the ability of Ang II to activate ERK2 in these cells. Controls for this study were the same cells expressing Jak2 via the addition of a Jak2 expression plasmid. In the cells expressing Jak2, Ang II induced a marked increase in ERK2 activity as measured by Western blot analysis and in vitro kinase assays. ERK2 activity returned to basal levels within 30 min. However, in the cells lacking Jak2, Ang II treatment resulted in ERK2 activation that did not return to basal levels until 120 min after ligand addition. Analysis of phosphatase gene expression revealed that Ang II induced mitogen-activated protein kinase phosphatase 1 (MKP-1) expression in cells expressing Jak2 but failed to induce MKP-1 expression in cells lacking Jak2. Therefore, our results suggest that Jak2 is not required for Ang II-induced ERK2 activation. Rather Jak2 is required for Ang II-induced ERK2 inactivation via induction of MKP-1 gene expression. PMID- 14551205 TI - Cell cycle-regulated phosphorylation of hamartin, the product of the tuberous sclerosis complex 1 gene, by cyclin-dependent kinase 1/cyclin B. AB - Tuberous sclerosis complex is a tumor suppressor gene syndrome whose manifestations can include seizures, mental retardation, and benign tumors of the brain, skin, heart, and kidneys. Hamartin and tuberin, the products of the TSC1 and TSC2 genes, respectively, form a complex and inhibit signaling by the mammalian target of rapamycin. Here, we demonstrate that endogenous hamartin is threonine-phosphorylated during nocodazole-induced G2/M arrest and during the G2/M phase of a normal cell cycle. In vitro assays showed that cyclin-dependent kinase 1 phosphorylates hamartin at three sites, one of which (Thr417) is in the hamartin-tuberin interaction domain. Tuberin interacts with phosphohamartin, and tuberin expression attenuates the phosphorylation of exogenous hamartin. Hamartin with alanine mutations in the three cyclin-dependent kinase 1 phosphorylation sites increased the inhibition of p70S6 kinase by the hamartin-tuberin complex. These findings support a model in which phosphorylation of hamartin regulates the function of the hamartin-tuberin complex during the G2/M phase of the cell cycle. PMID- 14551206 TI - Relationships between heme incorporation, tetramer formation, and catalysis of a heme-regulated phosphodiesterase from Escherichia coli: a study of deletion and site-directed mutants. AB - The heme-regulated phosphodiesterase (PDE) from Escherichia coli (Ec DOS) is a tetrameric protein composed of an N-terminal sensor domain (amino acids 1-201) containing two PAS domains (PAS-A, amino acids 21-84, and PAS-B, amino acids 144 201) and a C-terminal catalytic domain (amino acids 336-799). Heme is bound to the PAS-A domain, and the redox state of the heme iron regulates PDE activity. In our experiments, a H77A mutation and deletion of the PAS-B domain resulted in the loss of heme binding affinity to PAS-A. However, both mutant proteins were still tetrameric and more active than the full-length wild-type enzyme (140% activity compared with full-length wild type), suggesting that heme binding is not essential for catalysis. An N-terminal truncated mutant (DeltaN147, amino acids 148-807) containing no PAS-A domain or heme displayed 160% activity compared with full-length wild-type protein, confirming that the heme-bound PAS-A domain is not required for catalytic activity. An analysis of C-terminal truncated mutants led to mapping of the regions responsible for tetramer formation and revealed PDE activity in tetrameric proteins only. Mutations at a putative metal-ion binding site (His-590, His-594) totally abolished PDE activity, suggesting that binding of Mg2+ to the site is essential for catalysis. Interestingly, the addition of the isolated PAS-A domain in the Fe2+ form to the full-length wild-type protein markedly enhanced PDE activity (>5-fold). This activation is probably because of structural changes in the catalytic site as a result of interactions between the isolated PAS-A domain and that of the holoenzyme. PMID- 14551207 TI - The Akt-regulated forkhead transcription factor FOXO3a controls endothelial cell viability through modulation of the caspase-8 inhibitor FLIP. AB - FLICE-inhibitory protein (FLIP) is a homolog of caspase-8 that lacks catalytic activity and has been shown to be important in protecting endothelial cells from apoptosis. The serine/threonine kinase Akt/PKB was recently reported to promote FLIP expression in endothelial and tumor cells. Here we examined the role of the forkhead transcription factor FOXO3a, a downstream target of Akt, in controlling FLIP regulation in endothelial cells. FOXO3a nuclear translocation was regulated by Akt in human umbilical vein endothelial cells. Transduction of a nonphosphorylatable, constitutively active mutant of FOXO3a (TM-FOXO3a) led to the down-regulation of FLIP levels. Transduction with TM-FOXO3a also increased caspase-8 activity and promoted apoptosis in endothelial cells. Conversely, transduction of a dominant-negative mutant of FOXO3a up-regulated FLIP levels and protected endothelial cells from apoptosis under serum deprivation conditions. Restoration of intracellular FLIP blocked caspase-8 activation and inhibited apoptosis in TM-FOXO3a-transduced cells. These data suggest that FOXO3a is a downstream target of Akt in endothelial cells that can promote apoptosis via FLIP down-regulation and activation of the extrinsic apoptotic pathway. PMID- 14551208 TI - Cardiolipin is not required for Bax-mediated cytochrome c release from yeast mitochondria. AB - Cardiolipin (CL) is an inner mitochondrial membrane phospholipid that contributes to optimal mitochondrial function and is gaining widespread attention in studies of mitochondria-mediated apoptosis. Divergent hypotheses describing the role of CL in cytochrome c release and apoptosis have evolved. We addressed this controversy directly by comparing the spontaneous- and Bax-mediated cytochrome c release from mitochondria isolated from two strains of Saccharomyces cerevisiae: one lacking CL-synthase and therefore CL (DeltaCRD1) and the other, its corresponding wild type (WT). We demonstrated by liquid chromatography-mass spectrometry that the main yeast CL species [(16:1)2(18:1)2] differs in fatty acid composition from mammalian CL [(18:2)4], and we verified the absence of the yeast CL species in the DeltaCRD1 strain. We also demonstrated that the mitochondrial association of Bax and the resulting cytochrome c release is not dependent on the CL content of the yeast mitochondrial membranes. Bax inserted equally into both WT and DeltaCRD1 mitochondrial membranes under conditions that lead to the release of cytochrome c from both strains of yeast mitochondria. Furthermore, using models of synthetic liposomes and isolated yeast mitochondria, we found that cytochrome c was bound more "loosely" to the CL-deficient systems compared with when CL is present. These data challenge recent studies implicating that CL is required for Bax-mediated pore formation leading to the release of proteins from the mitochondrial intermembrane space. In contrast, they support our recently proposed two-step mechanism of cytochrome c release, which suggests that CL is required for binding cytochrome c to the inner mitochondrial membrane. PMID- 14551209 TI - Smad4 and beta-catenin co-activators functionally interact with lymphoid enhancing factor to regulate graded expression of Msx2. AB - Recent in vivo evidence suggests that Wnt signaling plays a central role in determining the fate of stem cells in the ectoderm and in the neural crest by modulating bone morphogenetic protein (BMP) levels, which, in turn, influence Msx gene expression. However, the molecular mechanism regulating the expression of the Msx genes as key regulators of cell fate has not been elucidated. Here we show in murine embryonic stem cells that BMP-dependent activation of Msx2 is mediated via the cooperative binding of Smad4 at two Smad binding elements and of lymphoid enhancing factor (Lef1) at two Lef1/TCF binding sites. Lef1 can synergize with Smad4 and Smad1 to activate Msx2 promoter, and this transcriptional complex is assembled on the endogenous promoter in response to BMP2. The Wnt/beta-catenin signaling pathway can activate Msx2 via the binding of Lef1 to its promoter and synergizes with BMP2 to activate Msx2 expression, possibly via enhanced recruitment of the p300/cAMP-response element-binding protein-binding protein co-factor. Interestingly, the Wnt/beta-catenin-dependent activation of Msx2 was defective in Smad4-deficient embryonic stem cells or when Smad binding elements were mutated but persisted in the presence of various BMP antagonists, indicating that Smad4 was involved in transducing the Wnt/beta catenin signals in the absence of a BMP autocrine loop. A chromatin immunoprecipitation analysis revealed that endogenous Smad4, but not Smad1, was part of the Lef1 transcriptional complex in response to beta-catenin activation, dismissing any implication of BMP signaling in this response. We propose that Wnt signaling pathway could dictate cell fate not only by modulating BMP levels but also by directly regulating cooperatively BMP-target genes. PMID- 14551210 TI - Cell cycle arrest and apoptosis induction by activator protein 2alpha (AP-2alpha) and the role of p53 and p21WAF1/CIP1 in AP-2alpha-mediated growth inhibition. AB - Activator protein 2alpha (AP-2alpha) is a sequence-specific DNA-binding transcription factor implicated in differentiation and transformation. In this study, we have made a replication-deficient recombinant adenovirus that expresses functional AP-2alpha (Ad-AP2). Cells infected with Ad-AP2 expressed induced levels of AP-2alpha protein, which bound to DNA in a sequence-specific manner and activated the AP-2-specific reporter 3X-AP2. Expression of AP-2alpha from Ad-AP2 inhibited cellular DNA synthesis and induced apoptosis. Ad-AP2 infection resulted in efficient inhibition of growth of cancer cells of six different types. In addition, prior expression of AP-2alpha increased the chemosensitivity of H460, a lung carcinoma cell line, to adriamycin (2.5-fold) and cisplatin (5-fold). Furthermore, the growth inhibition by AP-2alpha was found to be less efficient in the absence of p53 or p21, which correlated with reduced apoptosis in p53 null cells and lack of DNA synthesis inhibition in p21WAF1/CIP1 null cells by AP 2alpha, respectively. These results suggest that AP-2alpha inhibits the growth of cells by inducing cell cycle arrest and apoptosis and that the use of AP-2alpha should be explored as a therapeutic strategy either alone or in combination with chemotherapy. PMID- 14551211 TI - Separation-of-function mutants of yeast Ku80 reveal a Yku80p-Sir4p interaction involved in telomeric silencing. AB - The Saccharomyces cerevisiae Ku heterodimer comprising Yku70p and Yku80p is involved in telomere maintenance and DNA repair by the pathway of non-homologous end joining. It is also a key regulator of transcriptional silencing of genes placed in close proximity to telomeres. Here, we describe the identification of separation-of-function mutants of Yku80p that exhibit defects in silencing but not DNA repair and show that these mutations map to an evolutionarily conserved domain within Yku80p. Furthermore, we reveal that Yku80p interacts with the silent information regulator protein Sir4p and that this interaction is mediated by the N-terminal 200 amino acid residues of Sir4p. Notably, this interaction also requires the region of Yku80p that contains the sites of the silencing defective mutations. Finally, we show that these mutations impair the Yku80p Sir4p interaction and recruitment of Sir3p to telomeric regions in vivo. Taken together with other data, these findings indicate that the Yku80p-Sir4p interaction plays a vital role in the assembly of telomeric heterochromatin. PMID- 14551212 TI - Phosphorylations of cyclin-dependent kinase 2 revisited using two-dimensional gel electrophoresis. AB - To control the G1/S transition and the progression through the S phase, the activation of the cyclin-dependent kinase (CDK) 2 involves the binding of cyclin E then cyclin A, the activating Thr-160 phosphorylation within the T-loop by CDK activating kinase (CAK), inhibitory phosphorylations within the ATP binding region at Tyr-15 and Thr-14, dephosphorylation of these sites by cdc25A, and release from Cip/Kip family (p27kip1 and p21cip1) CDK inhibitors. To re-assess the precise relationship between the different phosphorylations of CDK2, and the influence of cyclins and CDK inhibitors upon them, we introduce here the use of the high resolution power of two-dimensional gel electrophoresis, combined to Tyr 15- or Thr-160-phosphospecific antibodies. The relative proportions of the potentially active forms of CDK2 (phosphorylated at Thr-160 but not Tyr-15) and inactive forms (non-phosphorylated, phosphorylated only at Tyr-15, or at both Tyr 15 and Thr-160), and their respective association with cyclin E, cyclin A, p21, and p27, were demonstrated during the mitogenic stimulation of normal human fibroblasts. Novel observations modify the current model of the sequential CDK2 activation process: (i) Tyr-15 phosphorylation induced by serum was not restricted to cyclin-bound CDK2; (ii) Thr-160 phosphorylation engaged the entirety of Tyr-15-phosphorylated CDK2 associated not only with a cyclin but also with p27 and p21, suggesting that Cip/Kip proteins do not prevent CDK2 activity by impairing its phosphorylation by CAK; (iii) the potentially active CDK2 phosphorylated at Thr-160 but not Tyr-15 represented a tiny fraction of total CDK2 and a minor fraction of cyclin A-bound CDK2, underscoring the rate-limiting role of Tyr-15 dephosphorylation by cdc25A. PMID- 14551213 TI - Constitutively active Galpha16 stimulates STAT3 via a c-Src/JAK- and ERK dependent mechanism. AB - The hematopoietic-specific Galpha16 protein has recently been shown to mediate receptor-induced activation of the signal transducer and activator of transcription 3 (STAT3). In the present study, we have delineated the mechanism by which Galpha16 stimulates STAT3 in human embryonic kidney 293 cells. A constitutively active Galpha16 mutant, Galpha16QL, stimulated STAT3-dependent luciferase activity as well as the phosphorylation of STAT3 at both Tyr705 and Ser727. Galpha16QL-induced STAT3 activation was enhanced by overexpression of extracellular signal-regulated kinase 1 (ERK1), but was inhibited by U0126, a Raf 1 inhibitor, and coexpression of the dominant negative mutants of Ras and Rac1. Inhibition of phospholipase Cbeta, protein kinase C, and calmodulin-dependent kinase II by their respective inhibitors also suppressed Galpha16QL-induced STAT3 activation. The involvement of tyrosine kinases such as c-Src and Janus kinase 2 and 3 (JAK2 and JAK3) in Galpha16QL-induced activation of STAT3 was illustrated by the combined use of selective inhibitors and dominant negative mutants. In contrast, c-Jun N-terminal kinase, p38 MAPK, RhoA, Cdc42, phosphatidylinositol 3 kinase, and the epidermal growth factor receptor did not appear to be required. Similar observations were obtained with human erythroleukemia cells, where STAT3 phosphorylation was stimulated by C5a in a PTX-insensitive manner. Collectively, these results highlight the important regulatory roles of the Ras/Raf/MEK/ERK and c-Src/JAK pathways on the stimulation of STAT3 by activated Galpha16. Demonstration of the involvement of different kinases in Galpha16QL-induced STAT3 activation supports the involvement of multiple signaling pathways in the regulation of transcription by G proteins. PMID- 14551214 TI - Remodeling of cardiolipin by phospholipid transacylation. AB - Mitochondrial cardiolipin (CL) contains unique fatty acid patterns, but it is not known how the characteristic molecular species of CL are formed. We found a novel reaction that transfers acyl groups from phosphatidylcholine or phosphatidylethanolamine to CL in mitochondria of rat liver and human lymphoblasts. Acyl transfer was stimulated by ADP, ATP, and ATP gamma S, but not by other nucleotides. Coenzyme A stimulated the reaction only in the absence of adenine nucleotides. Free fatty acids were not incorporated into CL under the same incubation condition. The transacylation required addition of exogenous CL or monolyso-CL, whereas dilyso-CL was not a substrate. Transacylase activity was decreased in lymphoblasts from patients with Barth syndrome (tafazzin deletion), and this was accompanied by drastic changes in the molecular composition of CL. In rat liver, where linoleic acid was the most abundant residue of CL, only linoleoyl groups were transferred into CL, but not oleoyl or arachidonoyl groups. We demonstrated complete remodeling of tetraoleoyl-CL to tetralinoleoyl-CL in rat liver mitochondria and identified the intermediates linoleoyl-trioleoyl-CL, dilinoleoyl-dioleoyl-CL, and trilinoleoyl-oleoyl-CL by high-performance liquid chromatography. The data suggest that CL is remodeled by acyl specific phospholipid transacylation and that tafazzin is an acyltransferase involved in this mechanism. PMID- 14551215 TI - The Kruppel-like factor epiprofin is expressed by epithelium of developing teeth, hair follicles, and limb buds and promotes cell proliferation. AB - We identified a cDNA clone for epiprofin, which is preferentially expressed in teeth, by differential hybridization using DNA microarrays from an embryonic day 19.5 mouse molar cDNA library. Sequence analysis revealed that this cDNA encodes a member of the Kruppel-like factor family containing three characteristic C2H2 type zinc finger motifs. The full-length cDNA was obtained by the 5' Cap capture method. Except for its 5'-terminal sequence, the epiprofin mRNA sequence is almost identical to the predicted sequence of Kruppel-like factor 14/Sp6 (specificity protein 6), which was previously identified in expressed sequence tag data bases and GenBank by an Sp1 zinc finger DNA-binding domain search (Scohy, S., Gabant, P., Van Reeth, T., Hertveldt, V., Dreze, P. L., Van Vooren, P., Riviere, M., Szpirer, J., and Szpirer, C. (2000) Genomics 70, 93-101). This sequence difference is due to differences in the assignment of the location of exon 1. In situ hybridization revealed that epiprofin mRNA is expressed by proliferating dental epithelium, differentiated odontoblast, and also hair follicle matrix epithelium. In addition, whole mount in situ hybridization showed transient expression of epiprofin mRNA in cells of the apical ectodermal ridge in developing limbs and the posterior neuropore. Transfection of an epiprofin expression vector revealed that this molecule is localized in the nucleus and promotes cell proliferation. Thus, epiprofin is a highly cell- and tissue specific nuclear protein expressed primarily by proliferating epithelial cells of teeth, hair follicles, and limbs that may function in the development of these tissues by regulating cell growth. PMID- 14551216 TI - Lipopolysaccharides indirectly stimulate apoptosis and global induction of apoptotic genes in fibroblasts. AB - Following Gram-negative bacterial infection there is a reduction in matrix producing cells. The goal of the present study was to examine the apoptotic effects of lipopolysaccharide (LPS) on fibroblastic cells and to investigate the role that the host response plays in this reaction. This was accomplished in vivo by subcutaneous inoculation of LPS in wild type and TNFR1(-/-)R2(-/-) mice. The direct effects of LPS on fibroblast apoptosis was studied in vitro with normal diploid human fibroblasts. The results indicate that LPS in vivo induces apoptosis of fibroblasts. By RNA profiling we demonstrated that LPS stimulates global expression of apoptotic genes and down-regulates anti-apoptotic genes. Fluorometric studies demonstrated that LPS in vivo significantly increased caspase-8 and caspase-3 activity and by use of specific inhibitors, the activation of caspase-3 was shown to be initiated by caspase-8 with no contribution from caspase-9. In vitro studies demonstrated that LPS did not induce apoptosis of fibroblasts, whereas tumor necrosis factor (TNF) did. In addition, the pattern of apoptotic gene expression induced by TNF in vitro was nearly identical to that induced by LPS in vivo, as measured by RNase protection assay. Moreover, pre-treatment of cells with TNF greatly enhanced apoptosis induced by a second stimulation with TNF 24 h later, suggesting that the global induction of pro-apoptotic genes was functionally significant. Thus, LPS acts to modulate the expression of a large number of genes that favor apoptosis of fibroblastic cells that is dependent upon activation of caspase-8 and is largely mediated by TNF. PMID- 14551217 TI - Transition state analysis of the coupling of drug transport to ATP hydrolysis by P-glycoprotein. AB - ATPase activity associated with P-glycoprotein (Pgp) is characterized by three drug-dependent phases: basal (no drug), drug-activated, and drug-inhibited. To understand the communication between drug-binding sites and ATP hydrolytic sites, we performed steady-state thermodynamic analyses of ATP hydrolysis in the presence and absence of transport substrates. We used purified human Pgp (ABCB1, MDR1) expressed in Saccharomyces cerevisiae (Figler, R. A., Omote, H., Nakamoto, R. K., and Al-Shawi, M. K. (2000) Arch. Biochem. Biophys. 376, 34-46) as well as Chinese hamster Pgp (PGP1). Between 23 and 35 degrees C, we obtained linear Arrhenius relationships for the turnover rate of hydrolysis of saturating MgATP in the presence of saturating drug concentrations (kcat), from which we calculated the intrinsic enthalpic, entropic, and free energy terms for the rate limiting transition states. Linearity of the Arrhenius plots indicated that the same rate-limiting step was being measured over the temperature range employed. Using linear free energy analysis, two distinct transition states were found: one associated with uncoupled basal activity and the other with coupled drug transport activity. We concluded that basal ATPase activity associated with Pgp is not a consequence of transport of an endogenous lipid or other endogenous substrates. Rather, it is an intrinsic mechanistic property of the enzyme. We also found that rapidly transported substrates bound tighter to the transition state and required fewer conformational alterations by the enzyme to achieve the coupling transition state. The overall rate-limiting step of Pgp during transport is a carrier reorientation step. Furthermore, Pgp is optimized to transport drugs out of cells at high rates at the expense of coupling efficiency. The drug inhibition phase was associated with low affinity drug-binding sites. These results are consistent with an expanded version of the alternating catalytic site drug transport model (Senior, A. E., Al-Shawi, M. K., and Urbatsch, I. L. (1995) FEBS Lett. 377, 285-289). A new kinetic model of drug transport is presented. PMID- 14551218 TI - Enhanced activity of serum and urinary hyaluronidases in streptozotocin-induced diabetic Wistar and GK rats. AB - Using streptozotocin-induced diabetic Wistar and GK rats as models of type 1 and type 2 diabetes, respectively, we investigated the changes in serum and urinary hyaluronidase activity with the pathological progress. The serum hyaluronidase levels of streptozotocin-induced rats started to increase on the third day after injection and thereafter maintained approximately threefold higher levels compared with control rats; those of GK rats were already higher ( approximately twofold) from the beginning of the experiment. The increases of serum hyaluronidase activity in both diabetic rats were similar to those of blood glucose level, indicating that diabetes mellitus was accompanied by enhanced activity of circulating hyaluronidase from the early phase of its development. In zymography, every serum from diabetic and control rats gave two hyaluronidase isomers, a major 73-kDa band (Hyal-1 type) and a minor 132-kDa band, suggesting that the increases in serum hyaluronidase activity were not due to the appearance of novel isomers. The hyaluronidase activity in 24-h urine of streptozotocin induced rats was 3-, 7-, and 11-fold higher at the 8th, 15th, and 18th week than that of control rats, respectively, and the urinary hyaluronidase activity of GK rats was not significantly different from controls. There was a good correlation between the urinary hyaluronidase activity and the albumin excretion. Thus the increase in urinary hyaluronidase activity may reflect enhanced glomerular permeability in streptozotocin-induced diabetic rats and may be a useful marker for diabetic nephropathy. Relative resistance to SDS-denaturation in zymography of rat serum and urinary hyaluronidases compared with human serum hyaluronidase are also shown. PMID- 14551219 TI - Structural and topological studies on the lipid-mediated assembly of a membrane associated lipomannan in Micrococcus luteus. AB - The biosynthesis of three mannolipids and the presence of a membrane-associated lipomannan in Micrococcus luteus (formerly Micrococcus lysodeikticus) were documented over 30 years ago. Structural and topological studies have been conducted to learn more about the possible role of the mannolipids in the assembly of the lipomannan. The major mannolipid has been purified and characterized as alpha-D-mannosyl-(1 --> 3)-alpha-D-mannosyl-(1 --> 3) diacylglycerol (Man2-DAG) by negative-ion electrospray-ionization multistage mass spectrometry (ESI-MSn). Analysis of the fragmentation patterns indicates that the sn-1 position is predominantly acylated with a 12-methyltetradecanoyl group and the sn-2 position is acylated with a myristoyl group. The lipomannan is shown to be located on the exterior face of the cytoplasmic membrane, and not exposed on the surface of intact cells, by staining of intact protoplasts with fluorescein isothiocyanate (FITC)-linked concanavalin A (Con A). When cell homogenates of M. luteus are incubated with GDP-[3H]mannose (GDP-Man), [3H]mannosyl units are incorporated into Man1-2-DAG, mannosylphosphorylundecaprenol (Man-P-Undec) and the membrane-associated lipomannan. The addition of amphomycin, an inhibitor of Man-P-Undec synthesis, had no effect on the synthesis of Man1-2-DAG, but blocked the incorporation of [3H]mannose into Man-P-Undec and consequently the lipomannan. These results strongly indicate that GDP-Man is the direct mannosyl donor for the synthesis of Man1-2-DAG, and that the majority of the 50 mannosyl units in the lipomannan are derived from Man-P-Undec. Protease-sensitivity studies with intact and lysed protoplasts indicate that the active sites of the mannosyltransferases catalyzing the formation of Man1-2-DAG and Man-P-Undec are exposed on the inner face, and the Man-P-Undec-mediated reactions occur on the outer surface of the cytoplasmic membrane. Based on all of these results, a topological model is proposed for the lipid-mediated assembly of the membrane bound lipomannan. PMID- 14551220 TI - Addition of beta1-6 GlcNAc branching to the oligosaccharide attached to Asn 772 in the serine protease domain of matriptase plays a pivotal role in its stability and resistance against trypsin. AB - beta1-6 GlcNAc branching, a product of N-acetylglucosaminyltransferase V (GnT-V), is a key structure that is associated with malignant transformations and cancer metastasis. Although a number of reports concerning tumor metastasis-related glycoproteins that contain beta1-6 GlcNAc branching have appeared, the precise function of beta1-6 GlcNAc branching on glycoproteins remains to be elucidated. We previously reported on the importance of beta1-6 GlcNAc branching on matriptase in terms of proteolytic degradation in tumor metastasis. We report here that matriptase purified from GnT-V transfectant (beta1-6 GlcNAc matriptase) binds strongly to L4-PHA, which preferentially recognizes beta1-6 GlcNAc branches of tri- or tetraantennary sugar chains, indicating that the isolated matriptase contains beta1-6 GlcNAc branching. The beta1-6 GlcNAc matriptase was resistant to autodegradation, as well as trypsin digestion, compared with matriptase purified from mock-transfected cells. Furthermore, N-glycosidase-F treatment of beta1-6 GlcNAc matriptase greatly reduced its resistance to degradation. An analysis of matriptase mutants that do not contain potential N-glycosylation sites clearly shows that the beta1-6 GlcNAc branching on N-glycans attached to Asn 772 in the serine protease domain plays a major role in trypsin resistance. This is the first example of a demonstration of a direct relationship between beta1-6 GlcNAc branching and a biological function at the protein level. PMID- 14551221 TI - Alteration of neural tissue structure by expression of polysialic acid induced by viral delivery of PST polysialyltransferase. AB - The expression of polysialic acid (PSA) on neural cell adhesion molecule (NCAM) is known to attenuate cell-cell interactions. During neural development the widespread expression of PSA-NCAM creates permissive conditions for the migration of neuronal and glial precursors and the guidance and targeting of axons. NCAM polysialylation can occur via either of two specific sialyltransferases, ST8SiaII (STX) and ST8SiaIV (PST), and the purpose of this study was to determine if retroviral delivery of either PST or STX could induce PSA expression in vivo and thereby alter tissue plasticity. Retroviruses expressing GFP-PST or GFP-STX were injected into embryonic retina, and development was evaluated by examining neuroepithelial structure, the expression of markers for specific cell types, cellular proliferation, and apoptosis. Chick retina was chosen because it down regulates PSA early in its development and has a highly stereotyped program of morphogenesis. Retroviral expression of PST induced PSA expression in retina and resulted in severe but localized alterations in retinal morphogenesis, including an early disruption of radial glial cell morphology, highly disorganized retinal layers, and invasion of pigmented cells into the neural retina. In contrast, retroviral delivery of STX did not induce PSA expression or affect morphogenesis. These findings demonstrate that expression of PSA is sufficient to promote morphological alterations in a relatively nonplastic neural tissue. PMID- 14551222 TI - Impaired coordination of nutrient intake and substrate oxidation in melanocortin 4 receptor knockout mice. AB - Mutations in the melanocortin-4 receptor (MC4R) are associated with obesity. The obesity syndrome observed in humans with MC4R haploinsufficiency is similar to that observed in MC4R knockout mice, including increased longitudinal growth, hyperphagia, and fasting hyperinsulinemia. For comparison with other commonly investigated models of obesity and insulin resistance, we have backcrossed Mc4r-/ mice into the C57BL/6J (B6) background. Female obese Mc4r-/- mice exhibit reduced energy expenditure and an attenuated increase in fatty acid (FA) oxidation after exposure to high-fat diets compared with obese Lepob/Lepob mice. The reduced energy expenditure and FA oxidation correlates with changes in hepatic gene expression. The expression of genes involved in FA oxidation increased in obese Lepob/Lepob mice compared with wild-type and obese Mc4r-/- mice. In contrast, a key lipogenic enzyme, FA synthase (FAS), is increased in obese Mc4r-/- mice compared with obese Lepob/Lepob mice. Hyperinsulinemia, increased FAS mRNA expression and hepatic steatosis appear to be secondary to obesity in B6 Mc4r-/- mice. However, Mc4r-/- mice in a mixed genetic background develop severe hepatic steatosis at an early age. This might suggest an important role of the MC4R in regulating liver FA metabolism that is masked on the B6 background. Interestingly, the 10- to 20-fold increase in liver triglyceride in the outbred strain of Mc4r-/- mice is not always associated with fasting hyperinsulinemia or increased FAS mRNA expression. This observation suggests that changes in liver secondary to triglyceride accumulation lead to hyperinsulinemia and increased hepatic FAS expression in Mc4r-/- mice. PMID- 14551223 TI - Gonadotropin-releasing hormone receptor-mediated growth suppression of immortalized LbetaT2 gonadotrope and stable HEK293 cell lines. AB - Continuous administration of GnRH analogs results in an inhibition of tumor growth that may be mediated in part by direct activation of GnRH receptors (GnRHRs) expressed on tumor cells. However, it is not fully understood how the GnRHR mediates these growth effects. This study aimed to determine how the presence or absence of this receptor in different cell types might affect the ability of GnRH to directly mediate growth effects. We demonstrate that continuous treatment with GnRH or a GnRH agonist (GnRHA) induces an antiproliferative effect in a gonadotrope-derived cell line (LbetaT2) and also in HEK293 cells stably expressing either the rat or human GnRHR. The antiproliferative effect was time and dose dependent and was verified using [3H]thymidine incorporation, light microscopy, and analysis of cell number. Inhibition was specifically mediated via the GnRHR, as cotreatment of the GnRHR expressing cell lines with a GnRH antagonist blocked the growth-suppressive effect induced by GnRHA treatment. Cell cycle analysis revealed that GnRHA treated HEK/GnRHR cell lines induced an accumulation of cells in the G2/M phase, whereas a G0/G1 arrest was observed in LbetaT2 cells. GnRHA treatment also caused a small, but significant, increase in apoptotic cells. This study provides evidence for a direct role for the GnRHR in mediating antiproliferative events in two cell systems, neither of which was derived from extrapituitary reproductive tumors. The ability to induce these effects, regardless of the cell system involved, has implications regarding the use of GnRH analogs for the treatment of endocrine-related disorders and tumors. PMID- 14551224 TI - Regulation of collagen synthesis in mouse skin fibroblasts by distinct angiotensin II receptor subtypes. AB - We examined the possibility of whether angiotensin (Ang) II type 1 (AT1) and type 2 (AT2) receptor stimulation differentially regulates collagen production in mouse skin fibroblasts. Both AT1 and AT2 receptors were expressed in neonatal skin fibroblasts prepared from wild-type mice to a similar degree, and the AT1a receptor was exclusively expressed as opposed to the AT1b receptor. In wild-type fibroblasts, Ang II increased collagen synthesis accompanied by an increase in expression of tissue inhibitor of metalloproteinase (TIMP)-1, and these increases were inhibited by valsartan, an AT1 receptor blocker, but augmented by PD123319, an AT2 receptor antagonist. Ang II decreased basal and IGF-I-induced collagen production and inhibited TIMP-1 expression in neonatal skin fibroblasts prepared from AT1a knockout (KO) mice. These Ang II-mediated inhibitory effects on collagen production and TIMP-1 expression observed in AT1a KO fibroblasts were attenuated by the addition of PD123319 or a tyrosine phosphatase inhibitor, sodium orthovanadate, but not affected by a serine/threonine phosphatase inhibitor, okadaic acid. Moreover, we demonstrated that transfection of a catalytically inactive, dominant negative SHP-1 (Src homology 2-containing protein-tyrosine phosphatase-1) mutant inhibited the Ang II-mediated inhibitory effect on both collagen synthesis and TIMP-1 expression in AT1a KO fibroblasts. These results suggest that AT1a receptor stimulation increases collagen production in skin fibroblasts at least in part due to the inhibition of collagen degradation via the increase in TIMP-1 expression, whereas AT2 receptor stimulation exerts inhibitory effects on TIMP-1 expression, which is mediated at least partially by the activation of SHP-1, thereby possibly inhibiting collagen production. PMID- 14551225 TI - Growth hormone (GH)-stimulated insulin-like growth factor I gene expression is mediated by a tyrosine phosphorylation pathway depending on C-terminal region of human GH receptor in human GH receptor-expressing Ba/F3 cells. AB - The signaling pathway of GH-stimulated IGF-I gene expression is still unclear, although it has been reported that the Janus kinase (JAK)-signal transducers and activators of transcription (STAT)5b pathway plays an important role in liver IGF I expression. In this study, the GH-dependent IGF-I gene expression and its intracellular signaling mechanism have been examined in mouse pro-B, Ba/F3 cells stably expressing human GH receptor (Ba/F3-hGHR). The IGF-I gene expression was stimulated by human GH (0.01-10 nm) in a dose-dependent fashion in Ba/F3-hGHR cells. The specific inhibitors for JAK2 remarkably suppressed the GH-induced IGF I gene expression, but MAPK or phosphatidylinositol 3 kinase-specific inhibitors failed to block the GH stimulation of the IGF-I gene expression. However, genistein, a nonspecific tyrosine kinase inhibitor that does not inhibit JAK2 and STAT5 phosphorylation, significantly suppressed the GH-induced IGF-I gene expression. Additionally, a Ba/F3-hGHR mutant that contained the truncated C terminal hGHR up to D351 showed no IGF-I gene expression in response to human GH. The D351 form normally has the GH-induced JAK/STAT5 tyrosine phosphorylation. These results suggest that the JAK-STAT5 pathway and the novel tyrosine phosphorylation pathway, dependent on signaling from the C-terminal region of hGHR, might be involved in the GH-stimulated IGF-I gene expression in Ba/F3 cells. PMID- 14551226 TI - In vitro and in vivo regulation of antioxidant response element-dependent gene expression by estrogens. AB - Understanding estrogen's regulation of phase II detoxification enzymes is important in explaining how estrogen exposure increases the risk of developing certain cancers. Phase II enzymes such as glutathione-S-transferases (GST) and quinone reductase protect against developing chemically induced cancers by metabolizing reactive oxygen species. Phase II enzyme expression is regulated by a cis-acting DNA sequence, the antioxidant response element (ARE). It has previously been reported that several antiestrogens, but not 17beta-estradiol, could regulate ARE-mediated gene transcription. Our goal was to determine whether additional estrogenic compounds could regulate ARE-mediated gene expression both in vitro and in vivo. We discovered that physiological concentrations (10 nm) of 17beta-estradiol repressed GST Ya ARE-dependent gene expression in vitro. Treatment with other endogenous and anti-, xeno-, and phytoestrogens showed that estrogen receptor/ARE signaling is ligand, receptor subtype, and cell type specific. Additionally, GST and quinone reductase activities were significantly lowered in a dose-dependent manner after 17beta-estradiol exposure in the uteri of mice. In conclusion, we have shown that 17beta-estradiol, and other estrogens, down-regulate phase II enzyme activities. We propose estrogen-mediated repression of phase II enzyme activities may increase cellular oxidative DNA damage that ultimately can result in the formation of cancer in some estrogen-responsive tissues. PMID- 14551227 TI - Norepinephrine increases glucose transport in brown adipocytes via beta3 adrenoceptors through a cAMP, PKA, and PI3-kinase-dependent pathway stimulating conventional and novel PKCs. AB - To identify the signaling pathways that mediate the adrenergic stimulation of glucose uptake in brown adipose tissue, we used mouse brown adipocytes in culture. The endogenous adrenergic neurotransmitter norepinephrine (NE) induced 2 deoxy-D-glucose uptake 3-fold in a concentration-dependent manner (pEC50 approximately 6.5). The uptake was abolished by high doses of propranolol. The NE effect was mimicked by isoprenaline (pEC50 approximately 6.9), BRL 37344 (pEC50 approximately 8.6), CL 316243 (pEC50 approximately 9.7) and CGP 12177 (pEC50 approximately 7.3) and was thus mediated by beta3-adrenergic receptors. The NE induced effect on 2-deoxy-D-glucose uptake was mediated by adenylyl cyclase and cAMP because responses were inhibited by the adenylyl cyclase inhibitor 2',5' dideoxyadenosine and the protein kinase A inhibitor 4-cyano-3-methylisoquinoline. Cholera toxin and 8-bromoadenosine cAMP were both able to increase 2-deoxy-D glucose uptake. Involvement of other adrenergic signaling pathways (alpha1-and alpha2-adrenergic receptors) were excluded. The phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002, abolished beta-adrenergic- or 8-bromoadenosine cAMP stimulated 2-deoxy-D-glucose uptake, demonstrating that a cAMP-dependent PI3K mediated pathway is positively connected to glucose uptake. Inhibition of the beta-adrenergically stimulated response with protein kinase C (PKC) inhibitors (Go 6983, which inhibits (alpha, beta, gamma), (delta), and (zeta) isoforms and Ro-31-8220, which inhibits (alpha, beta1, beta2, gamma) and (epsilon) but not atypical isoforms) indicated that cAMP-mediated glucose uptake is stimulated via conventional and novel PKCs. These results demonstrate that adrenergic stimulation, through beta3-adrenergic receptors/cAMP/protein kinase A, recruits a PI3K pathway stimulating conventional and novel PKCs, which mediate glucose uptake in brown adipocytes. PMID- 14551229 TI - Generation of a phenotypic array of hypothalamic neuronal cell models to study complex neuroendocrine disorders. AB - Knowledge of how the brain achieves its diverse central control of basic physiology is severely limited by the virtual absence of appropriate cell models. Isolation of clonal populations of unique peptidergic neurons from the hypothalamus will facilitate these studies. Herein we describe the mass immortalization of mouse primary hypothalamic cells in monolayer culture, resulting in the generation of a vast representation of hypothalamic cell types. Subcloning of the heterogeneous cell populations resulted in the establishment of 38 representative clonal neuronal cell lines, of which 16 have been further characterized by analysis of 28 neuroendocrine markers. These cell lines represent the first available models to study the regulation of neuropeptides associated with the control of feeding behavior, including neuropeptide Y, ghrelin, urocortin, proopiomelanocortin, melanin-concentrating hormone, neurotensin, proglucagon, and GHRH. Importantly, a representative cell line responds appropriately to leptin stimulation and results in the repression of neuropeptide Y gene expression. These cell models can be used for detailed molecular analysis of neuropeptide gene regulation and signal transduction events involved in the direct hormonal control of unique hypothalamic neurons, not yet possible in the whole brain. Such studies may contribute information necessary for the strategic design of therapeutic interventions for complex neuroendocrine disorders, such as obesity. PMID- 14551228 TI - Ghrelin and des-octanoyl ghrelin promote adipogenesis directly in vivo by a mechanism independent of the type 1a growth hormone secretagogue receptor. AB - Ghrelin promotes fat accumulation, despite potent stimulation of the lipolytic hormone, GH. The function of the major circulating isoform of ghrelin, des octanoyl ghrelin, is unclear, because it does not activate the GH secretagogue receptor (GHS-R1a) and lacks the endocrine activities of ghrelin. We have now addressed these issues by infusing ghrelin, des-octanoyl ghrelin, or synthetic GHS-R1a agonists into three rat models with moderate, severe, or total GH deficiency. We show that in the context of significant GH secretion, the adipogenic effect of systemic ghrelin infusion is pattern dependent. However, this adipogenic action is not mediated by the pituitary hormones. Using a novel unilateral local infusion strategy, we demonstrate that ghrelin promotes bone marrow adipogenesis in vivo by a direct peripheral action. Surprisingly, this effect was also observed with des-octanoyl ghrelin, whereas a potent synthetic GHS-R1a agonist was ineffective. Thus, these adipogenic effects are mediated by a receptor other than GHS-R1a. This is the first in vivo demonstration of a direct adipogenic effect of des-octanoyl ghrelin, a major circulating form of ghrelin that lacks GH-releasing activity. We suggest that the ratio of ghrelin and des octanoyl ghrelin production could help regulate the balance between adipogenesis and lipolysis in response to nutritional status. PMID- 14551230 TI - Differential colocalization of Islet-1 and estrogen receptor alpha in the murine preoptic area and hypothalamus during development. AB - Estrogen receptor (ER) expression and regulation is vital to the correct functioning of the neuroendocrine brain. Islet-1 (Isl-1) is a LIM homeodomain containing transcription factor that has been implicated in neuronal differentiation, is located in the hypothalamus, and can alter ER function in vitro. We have determined that Isl-1 is localized in several regions of the hypothalamus, including the ER rich areas of the ventromedial nucleus (VMH), the preoptic area, and the anterior hypothalamus. Using double-label immunocytochemistry, we examined the overlap between immunoreactive ERalpha and Isl-1 in these different hypothalamic brain regions. In the developing brain, almost 100% of VMH cells that contain immunoreactive ERalpha also contain Isl-1. However, in older animals, the percentage of double-label cells decreased below 70%. This change is due to a decrease in the number of cells containing Isl-1, because there was no difference in the number of ERalpha-containing cells. By contrast, in more anterior regions of the hypothalamus, cells containing both Isl 1 and ERalpha were less common, with the two populations adjacent to each other, rather than overlapping. These data suggest that, although Isl-1 and ERalpha can interact, they are not always found in the same cells and that regulation of ERalpha function is not under the same control in the VMH, preoptic area, and the anterior hypothalamus. PMID- 14551231 TI - Endocrine and metabolic effects of growth hormone (GH) compared with GH-releasing peptide, thyrotropin-releasing hormone, and insulin infusion in a rabbit model of prolonged critical illness. AB - Treatment with recombinant human GH (rhGH) increases the mortality of critical illness. We postulated that combined GH-releasing peptide-2 (GHRP-2), TRH, and insulin infusion is a less toxic anabolic strategy through a putative inability to overstimulate the GH axis and a capacity to normalize thyroid hormone concentrations while foregoing excessive hyperglycemia. Burn-injured, parenterally fed, New Zealand White rabbits were randomized to receive 4-d treatment with saline (n=8); 60 microg/kg.h GHRP-2 and 60 microg/kg.h TRH, i.v. (n=9); or 3.5 mg/kg rhGH, s.c. (n=7). In the GHRP-2+TRH group, insulin was adjusted to maintain blood glucose below 180 mg/dl. Endocrine function and biochemical organ system function markers were studied. Animals were killed for assay of deiodinase activity in snap-frozen liver. Mortality, organ system function, hyperglycemia, and insulin requirement were equal in the three groups. GHRP-2+TRH increased pulsatile rabbit GH (rGH) and TSH release on d 1. After 4 d, rGH secretion and T4 and T3 concentrations were elevated, with a significant increase in hepatic activity of type 1 deiodinase and a decrease in type 3 deiodinase. Exogenous rhGH suppressed endogenous rGH secretion and increased IGF I more than GHRP-2+TRH without altering thyroid hormone levels. Unlike GHRP 2+TRH, rhGH down-regulated liver type 3 deiodinase and did not affect type 1 deiodinase. We conclude that in experimentally induced critical illness, GHRP 2+TRH reactivated the GH and TSH axes and altered liver deiodinase activity, driving T4 to T3 conversion. In contrast to the human model, high dose rhGH was not rapidly lethal in this rabbit model. Whether this is explained by lack of rhGH-induced insulin resistance and hyperglycemia remains unclear. PMID- 14551232 TI - Regulation of Sertoli cell number and activity by follicle-stimulating hormone and androgen during postnatal development in the mouse. AB - The roles of FSH and androgen in the postnatal development of Sertoli cell number and function have been investigated using mice that lack FSH (FSHbetaKO), FSH receptors (FSHRKO), or androgen receptors (Tfm). At birth and d 5, Sertoli cell number was normal in FSHRKO and FSHbetaKO mice, but was significantly reduced on d 20 and in adulthood. In contrast, Sertoli cell number was reduced at birth in Tfm mice and remained significantly less than normal up to adulthood. Sertoli cell activity was determined through measurement of 11 different mRNA transcript levels. From birth to adulthood, the expression of most transcripts increased, with a significant rise occurring between d 5 and 10. In animals lacking FSH stimulation, mRNA expression (measured per Sertoli cell) was largely normal on d 5, but was reduced in seven transcripts on d 20 and in five transcripts at adulthood. In Tfm mice two transcripts showed reduced expression on d 5, and four were reduced on d 20, although expression in adult Tfm mice did not differ from that in normal cryptorchid controls. The results show that 1) testosterone, but not FSH, is required for Sertoli cell proliferation during fetal and early neonatal life; 2) FSH and testosterone both regulate the late stages of Sertoli cell proliferation; 3) FSH has a general trophic effect on Sertoli cell activity in the pubertal and adult mouse; and 4) androgens are required for specific transcript expression during prepubertal development. Specific effects of androgens were not seen in the adult, although these may be masked by the effects of cryptorchidism. PMID- 14551233 TI - Hepatocyte growth factor gene therapy for pancreatic islets in diabetes: reducing the minimal islet transplant mass required in a glucocorticoid-free rat model of allogeneic portal vein islet transplantation. AB - Islet transplantation for diabetes is limited by the availability of human islet donors. Hepatocyte growth factor (HGF) is a potent beta-cell mitogen and survival factor and improves islet transplant outcomes in a murine model. However, the murine model employs renal subcapsular transplant and immunodeficient mice, features not representative of human islet transplantation protocols. Therefore, we have developed a more rigorous, marginal-mass rat islet transplant model that more closely resembles human islet transplantation protocols: islet donors are allogeneic Lewis islets; recipients are normal Sprague Dawley rats; islets are delivered intraportally; and immunosuppression is accomplished using the same immunosuppressants employed by the Edmonton group. We demonstrate that 1) surprisingly, the Edmonton immunosuppression regimen induces marked insulin resistance and beta-cell toxicity in rats, 2) adenovirus does not adversely affect islet transplant outcomes, 3) the Edmonton immunosuppressants may delay or block rejection of adenovirally transduced islets, and more importantly, 4) pretransplant islet adenoviral gene therapy with HGF markedly improves islet transplant outcomes, 5) this enhanced function persists for months, and 6) HGF enhances islet function and survival even in the setting of immunosuppressant induced insulin resistance and beta-cell toxicity. This approach may enhance islet transplantation outcomes in humans. PMID- 14551234 TI - Endotoxin inhibits the surge secretion of gonadotropin-releasing hormone via a prostaglandin-independent pathway. AB - Immune/inflammatory challenges, such as bacterial endotoxin, disrupt gonadotropin secretion and ovarian cyclicity. We previously determined that endotoxin can block the estradiol-induced LH surge in the ewe. Here, we investigated mechanisms underlying this suppression. First, we tested the hypothesis that endotoxin blocks the estradiol-induced LH surge centrally, by preventing the GnRH surge. Artificial follicular phases were created in ovariectomized ewes, and either endotoxin or vehicle was administered together with a surge-inducing estradiol stimulus. In each ewe in which endotoxin blocked the LH surge, the GnRH surge was also blocked. Given this evidence that endotoxin blocks the estradiol-induced LH surge at the hypothalamic level, we began to assess underlying central mechanisms. Specifically, in view of the prior demonstration that prostaglandins mediate endotoxin-induced suppression of pulsatile GnRH secretion in ewes, we tested the hypothesis that prostaglandins also mediate endotoxin-induced blockade of the surge. The prostaglandin synthesis inhibitor flurbiprofen was delivered together with endotoxin and the estradiol stimulus. Although flurbiprofen abolished endotoxin-induced fever, which is a centrally generated, prostaglandin mediated response, it failed to reverse blockade of the LH surge. Collectively, these results indicate endotoxin blocks the LH surge centrally, suppressing GnRH secretion via a mechanism not requiring prostaglandins. This contrasts with the suppressive effect of endotoxin on GnRH pulses, which requires prostaglandins as intermediates. PMID- 14551235 TI - Lignin: genetic engineering and impact on pulping. AB - Lignin is a major component of wood, the most widely used raw material for the production of pulp and paper. Although the biochemistry and molecular biology underpinning lignin production are better understood than they are for the other wood components, recent work has prompted a number of re-evaluations of the lignin biosynthetic pathway. Some of the work on which these revisions have been based involved the investigation of transgenic plants with modified lignin biosynthesis. In addition to their value in elucidating the lignin biosynthetic pathway, such transgenic plants are also being produced with the aim of improving plant raw materials for pulp and paper production. This review describes how genetic engineering has yielded new insights into how the lignin biosynthetic pathway operates and demonstrates that lignin can be improved to facilitate pulping. The current technologies used to produce paper are presented in this review, followed by a discussion of the impact of lignin modification on pulp production. Fine-tuned modification of lignin content, composition, or both is now achievable and could have important economic and environmental benefits. PMID- 14551236 TI - Nitrogen fixation: the mechanism of the Mo-dependent nitrogenase. AB - This review focuses on recent developments elucidating the mechanism of the Mo dependent nitrogenase. This enzyme, responsible for the majority of biological nitrogen fixation, is composed of two component proteins called the MoFe protein and the Fe protein. Recent progress in understanding the mechanism of this enzyme has focused on elucidating the structures of the active site metal clusters and of the proteins, understanding substrate interactions with the active site, defining the flow of electron transfer between the metal clusters, and defining the various roles of MgATP hydrolysis. PMID- 14551237 TI - Rapid effect of 3-hydroxy-3-methylglutaryl coenzyme a reductase inhibition on coronary endothelial function. AB - Treatment with 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) decreases cardiovascular event rates in hypercholesterolemic patients. Whether statins exert effects within 24 hours on the coronary vasculature in patients with endothelial dysfunction has not been elucidated. Twenty-seven patients with stable angina pectoris and average low-density lipoprotein cholesterol concentrations of 138+/-9 mg/dL at baseline were allocated to treatment with placebo (14 patients) or 40 mg/d pravastatin (13 patients) in a randomized, double-blind, prospective trial. Coronary endothelial function was assessed before and 24 hours after single treatment by quantitative coronary angiography during intracoronary infusion of nitroglycerin or increasing concentrations of acetylcholine (0.01, 0.1, and 1 micromol/L). Coronary blood flow reserve was measured by Doppler velocimetry during adenosine infusion. Intracoronary acetylcholine infusion induced abnormal vasoconstriction in both groups before treatment, indicating coronary endothelial dysfunction. Treatment with a single oral 40-mg dose of pravastatin significantly attenuated acetylcholine-mediated vasoconstriction after 24 hours (mean+/-SE decrease in luminal diameter before and after treatment: 0.01 micromol/L, 6.1+/-2.2% versus 3.0+/-1.2%; 0.1 micromol/L, 15.6+/-2.6% versus 7.4+/-1.8%; P<0.05; 1 micromol/L, 22.9+/-2.9% versus 13.2+/-2.6%; P<0.05). There was no significant difference in the response to acetylcholine in the placebo group (8.1+/-2.4% versus 9.7+/-2.4%, 16.1+/-2.9% versus 16.8+/-3.2%, and 21.4+/-3.9% versus 23.3+/-4.2%). The response to nitroglycerin infusion was not altered in both groups. Increase in coronary blood flow in response to adenosine and coronary flow reserve remained unchanged during placebo and statin treatment. Serum concentrations of blood lipids and high-sensitive C-reactive protein were not significantly altered after 24 hours in response to placebo or pravastatin therapy. Statin treatment improves endothelium-dependent coronary vasomotion within 24 hours in the absence of significant cholesterol reduction. The full text of this article is available online at http://www.circresaha.org. PMID- 14551238 TI - Contrasting roles of NADPH oxidase isoforms in pressure-overload versus angiotensin II-induced cardiac hypertrophy. AB - Increased production of reactive oxygen species (ROS) is implicated in the development of left ventricular hypertrophy (LVH). Phagocyte-type NADPH oxidases are major cardiovascular sources of ROS, and recent data indicate a pivotal role of a gp91phox-containing NADPH oxidase in angiotensin II (Ang II)-induced LVH. We investigated the role of this oxidase in pressure-overload LVH. gp91phox-/- mice and matched controls underwent chronic Ang II infusion or aortic constriction. Ang II-induced increases in NADPH oxidase activity, atrial natriuretic factor (ANF) expression, and cardiac mass were inhibited in gp91phox-/- mice, whereas aortic constriction-induced increases in cardiac mass and ANF expression were not inhibited. However, aortic constriction increased cardiac NADPH oxidase activity in both gp91phox-/- and wild-type mice. Myocardial expression of an alternative gp91phox isoform, Nox4, was upregulated after aortic constriction in gp91phox-/- mice. The antioxidant, N-acetyl-cysteine, inhibited pressure-overload-induced LVH in both gp91phox-/- and wild-type mice. These data suggest a differential response of the cardiac Nox isoforms, gp91phox and Nox4, to Ang II versus pressure overload. PMID- 14551239 TI - Two-pore domain K channel, TASK-1, in pulmonary artery smooth muscle cells. AB - Pulmonary vascular tone is strongly influenced by the resting membrane potential of smooth muscle cells, depolarization promoting Ca2+ influx, and contraction. The resting potential is determined largely by the activity of K+-selective ion channels, the molecular nature of which has been debated for some time. In this study, we provide strong evidence that the two-pore domain K+ channel, TASK-1, mediates a noninactivating, background K+ current (IKN), which sets the resting membrane potential in rabbit pulmonary artery smooth muscle cells (PASMCs). TASK 1 mRNA was found to be present in PASMCs, and the membranes of PASMCs contained TASK-1 protein. Both IKN and the resting potential were found to be exquisitely sensitive to extracellular pH, acidosis inhibiting the current and causing depolarization. Moreover, IKN and the resting potential were enhanced by halothane (1 mmol/L), inhibited by Zn2+ (100 to 200 micromol/L) and anandamide (10 micromol/L), but insensitive to cytoplasmic Ca2+. These properties are all diagnostic of TASK-1 channels and add to previously identified features of IKN that are shared with TASK-1, such as inhibition by hypoxia, low sensitivity to 4 aminopyridine and quinine and insensitivity to tetraethylammonium ions. It is therefore concluded that TASK-1 channels are major contributors to the resting potential in pulmonary artery smooth muscle. They are likely to play an important role in mediating pulmonary vascular responses to changes in extracellular pH, and they could be responsible for the modulatory effects of pH on hypoxic pulmonary vasoconstriction. PMID- 14551240 TI - C-terminal truncation of cardiac troponin I causes divergent effects on ATPase and force: implications for the pathophysiology of myocardial stunning. AB - Myocardial stunning is a form of reversible myocardial ischemia/reperfusion injury associated with systolic and diastolic contractile dysfunction. In the isolated rat heart model, myocardial stunning is characterized by specific C terminal proteolysis of the myofilament protein, troponin I (cTnI) that yields cTnI1-193. To determine the effect of this particular C-terminal truncation of cTnI, without the confounding factor of other stunning-induced protein modifications, a series of solution biochemical assays has been undertaken using the human homologue of mouse/rat cTnI1-193, cTnI1-192. Affinity chromatography and actin sedimentation experiments detected little, or no, difference between the binding of cTnI (cTnI1-209) and cTnI1-192 to actin-tropomyosin, troponin T, or troponin C. Both cTnI and cTnI1-192 inhibit the actin-tropomyosin-activated ATPase activity of myosin subfragment 1 (S1), and this inhibition is released by troponin C in the presence of Ca2+. However, cTnI1-192, when reconstituted as part of the troponin complex (cTn1-192), caused a 54+/-11% increase in the maximum Ca2+-activated actin-tropomyosin-S1 ATPase activity, compared with troponin reconstituted with cTnI (cTn). Furthermore, cTn1-192 increased Ca2+ sensitivity of both the actin-tropomyosin-activated S1 ATPase activity and the Ca2+-dependent sliding velocity of reconstituted thin filaments, in an in vitro motility assay, compared with cTn. In an in vitro force assay, the actin tropomyosin filaments bearing cTn1-192 developed only 76+/-4% (P<0.001) of the force obtained with filaments composed of reconstituted cTn. We suggest that cTnI proteolysis may contribute to the pathophysiology of myocardial stunning by altering the Ca2+-sensing and chemomechanical properties of the myofilaments. PMID- 14551241 TI - Does nitric oxide mediate the vasodilator activity of nitroglycerin? AB - Nitroglycerin (glyceryl trinitrate, GTN) relaxes blood vessels primarily via activation of the soluble guanylyl cyclase (sGC)/cGMP/cGMP-dependent protein kinase (cGK-I) pathway. Although the precise mechanism of sGC activation by GTN in the vascular wall is unknown, the mediatory role of nitric oxide (NO) has been postulated. We tested the GTN/NO hypothesis in different types of isolated rat and rabbit blood vessels using two novel approaches: (1) EPR spin trapping using colloid Fe(DETC)2 and (2) analysis of cGK-I-dependent phosphorylation of the vasodilator-stimulated phosphoprotein at Ser239 (P-VASP). For comparison, another organic nitrate, isosorbide dinitrate (ISDN), and endothelium-dependent vasodilator, calcium ionophore A23187, were tested. We found a marked discrepancy between GTN's strong vasoactivity (vasodilation and augmentation of P-VASP) and its poor NO donor properties. In aortas precontracted with phenylephrine, GTN, ISDN, and A23187 induced nearly full relaxations (>80%) and doubling of vascular P-VASP content at concentrations of 100 nmol/L, 100 micromol/L, and 1 micromol/L, respectively. GTN applied in vasorelaxant concentrations (10 to 1000 nmol/L) did not significantly increase the basal vascular NO production, in contrast to ISDN and A23187. The absence of GTN-derived NO was confirmed in rabbit vena cava and renal artery. A significant increase in vascular NO formation was observed only at suprapharmacological GTN concentrations (>10 micromol/L). The concentration dependency of NO formation from GTN was comparable to that of ISDN, although the latter exhibits 100-folds lower vasorelaxant potency. We conclude that GTN activates the sGC/cGMP/cGK-I pathway and induces vasorelaxation without intermediacy of the free radical NO. The full text of this article is available online at http://www.circresaha.org. PMID- 14551242 TI - Downregulation of the BK channel beta1 subunit in genetic hypertension. AB - The molecular mechanisms underlying increased arterial tone during hypertension are unclear. In vascular smooth muscle, localized Ca2+ release events through ryanodine-sensitive channels located in the sarcoplasmic reticulum (Ca2+ sparks) activate large-conductance, Ca2+-sensitive K+ (BK) channels. Ca2+ sparks and BK channels provide a negative feedback mechanism that hyperpolarizes smooth muscle and thereby opposes vasoconstriction. In this study, we examined Ca2+ sparks and BK channel function in Wistar-Kyoto (WKY) rats with borderline hypertension and in spontaneously hypertensive rats (SHR), a widely used genetic model of severe hypertension. We found that the amplitude of spontaneous BK currents in WKY and SHR cells were smaller than in normotensive cells even though Ca2+ sparks were of similar magnitude. BK channels in WKY and SHR cells were less sensitive to physiological changes in intracellular Ca2+ than normotensive cells. Our data indicate that decreased expression of the BK channel beta1 subunit underlies the lower Ca2+ sensitivity of BK channels in SHR and WKY myocytes. We conclude that the lower expression of the beta1 subunit during genetic borderline and severe hypertension reduced BK channel activity by decreasing the sensitivity of these channels to physiological changes in Ca2+. These results support the view that changes in the molecular composition of BK channels may be a fundamental event contributing to the development of vascular dysfunction during hypertension. PMID- 14551243 TI - Cholesterol depletion impairs vascular reactivity to endothelin-1 by reducing store-operated Ca2+ entry dependent on TRPC1. AB - The reactivity of the vascular wall to endothelin-1 (ET-1) is influenced by cholesterol, which is of possible importance for the progression of atherosclerosis. To elucidate signaling steps affected, the cholesterol acceptor methyl-beta-cyclodextrin (mbetacd, 10 mmol/L) was used to manipulate membrane cholesterol and disrupt caveolae in intact rat arteries. In endothelium-denuded caudal artery, contractile responsiveness to 10 nmol/L ET-1 (mediated by the ETA receptor) was reduced by mbetacd and increased by cholesterol. Neither ligand binding nor colocalization of ETA and caveolin-1 was affected by mbetacd. Ca2+ inflow via store-operated channels after depletion of intracellular Ca2+ stores was reduced in mbetacd-treated caudal arteries, as shown by Mn2+ quench rate and intracellular [Ca2+] response. Expression of TRPC1, 3, and 6 was detected by reverse transcriptase-polymerase chain reaction, and colocalization of TRPC1 with caveolin-1 was reduced by mbetacd, as seen by immunofluorescence. Part of the contractile response to ET-1 was inhibited by Ni2+ (0.5 mmol/L) and by a TRPC1 blocking antibody. In the basilar artery, exhibiting less store-operated channel activity than the caudal artery, ET-1-induced contractions were insensitive to the TRPC1 blocking antibody and to mbetacd. Increased store-operated channel activity in basilar arteries after organ culture correlated with increased sensitivity of ET-1 contraction to mbetacd. These results suggest that cholesterol influences vascular reactivity to ET-1 by affecting the caveolar localization of TRPC1. PMID- 14551244 TI - Developmental changes in ventricular diastolic function correlate with changes in ventricular myoarchitecture in normal mouse embryos. AB - Both genetic and epigenetic factors, such as abnormal hemodynamics, affect cardiac morphogenesis and the pathogenesis of congenital heart disease. Diastolic function is an important determinant of cardiac function, and tools for evaluating diastolic function in the embryo would be very valuable for assessment of cardiac performance. Using histological measurements of ventricular myoarchitecture, Doppler assessment of ventricular inflow velocities, and direct measurement of ventricular pressure, we investigated developmental changes of ventricular diastolic function in the mouse embryos from embryonic days 9.5 to 19.5. Regression analysis showed that peak velocity of A wave (an index of passive compliance) correlated with the area of trabecular myocardium in right ventricle (RV) (r2=0.92, P<0.0001) and left ventricle (LV) (r2=0.93, P<0.0001). Peak velocity of E wave (an index of active relaxation) exponentially correlated with the area of compact myocardium in RV (r2=0.98, P<0.0001) and LV (r2=0.97, P<0.0001). We used these techniques to analyze FOG-2 null embryos. FOG-2 null embryos had thin compact myocardium, higher EDP and E/A ratio, smaller -dP/dt, and diminished sucking pressure than wild-type littermates, indicating that decreased ventricular diastolic function might be the primary cause of embryonic lethality. In conclusion, during embryogenesis the development of compact myocardium tightly regulates the development of ventricular distensibility. Our study in normal mice forms the basis for future studies of embryonic cardiac function in genetically manipulated mice with abnormalities of the cardiovascular system. PMID- 14551245 TI - Treatment with insulin uncovers the motogenic capacity of nitric oxide in aortic smooth muscle cells: dependence on Gab1 and Gab1-SHP2 association. AB - Contrary to the antimotogenic effect of NO in dedifferentiated vascular smooth muscle cells (VSMCs), we have reported that NO stimulates the motility of differentiated cultured VSMC isolated from adult rats. This process involves upregulation of protein tyrosine phosphatase SHP2, followed by downregulation of RhoA activity. In the present study, we tested the hypothesis that insulin alters the motogenic phenotype of cultured rat aortic smooth muscle cells exposed to NO from inhibition to stimulation of cell motility. We demonstrate for the first time that NO stimulates the motility of VSMCs cultured for several days in the presence but not the absence of insulin. Moreover, we show that NO blocks PDGF induced cell motility in insulin-naive but not in insulin-treated cells. We also demonstrate that the scaffold adapter protein Gab1, considered a physiological activator of protein tyrosine phosphatase SHP2, increases cell motility in the presence but not the absence of insulin. In cells cultured in the presence of insulin, overexpression of Gab1 mimics, whereas a dominant-negative allele of Gab1 (Gab1YF) blocks, the motility-stimulatory effect of NO. Cotransfection experiments with dominant-negative Gab1 and wild-type SHP2 or wild-type Gab1 and dominant-negative SHP2 indicate that the two proteins work together as a functional unit to induce motility. Because chronic insulin can increase the levels of phosphatidylinositol 3 (PI3) kinase in several models of hyperinsulinemia, we also tested the potential involvement of this enzyme in mechanisms leading to increased cell motility. We found that the motogenic effect of NO, Gab1, and SHP2 was blocked by the selective PI3 kinase inhibitor LY294002, suggesting a requirement of PI3 kinase in mediating motogenesis. These observations may be relevant to molecular mechanisms related to the pathogenesis of vascular disease in hyperinsulinemic diabetes. The full text of this article is available online at http://www.circresaha.org. PMID- 14551246 TI - Apoptosis signal-regulating kinase 1 plays a pivotal role in angiotensin II induced cardiac hypertrophy and remodeling. AB - Multiple lines of evidence establish that angiotensin II (Ang II) induces not only hypertension but also directly contributes to cardiac diseases. Apoptosis signal-regulating kinase 1 (ASK1), one of mitogen-activated protein kinase kinase kinases, plays a key role in stress-induced cellular responses. However, nothing is known about the role of ASK1 in cardiac hypertrophy and remodeling in vivo. In this study, by using mice deficient in ASK1 (ASK1-/- mice), we investigated the role of ASK1 in cardiac hypertrophy and remodeling induced by Ang II. Left ventricular (LV) ASK1 was activated by Ang II infusion in wild-type mice, which was mediated by angiotensin II type 1 receptor and superoxide. Although Ang II induced hypertensive effect was comparable to wild-type and ASK1-/- mice, LV ASK1 activation by Ang II was not detectable in ASK1-/- mice, and p38 and c-Jun N terminal kinase (JNK) activation was lesser in ASK-/- mice than in wild-type mice. Elevation of blood pressure by continuous Ang II infusion was comparable between ASK1-/- and wild-type mice. However, Ang II-induced cardiac hypertrophy and remodeling, including cardiomyocyte hypertrophy, cardiac hypertrophy-related mRNA upregulation, cardiomyocyte apoptosis, interstitial fibrosis, coronary arterial remodeling, and collagen gene upregulation, was significantly attenuated in ASK1-/- mice compared with wild-type mice. These results provided the first in vivo evidence that ASK1 is the critical signaling molecule for Ang II-induced cardiac hypertrophy and remodeling. Thus, ASK1 is proposed to be a potential therapeutic target for cardiac diseases. PMID- 14551247 TI - Schizosaccharomyces pombe Rdh54 (TID1) acts with Rhp54 (RAD54) to repair meiotic double-strand breaks. AB - We report the characterization of rdh54+, the second fission yeast Schizosaccharomyces pombe Rad54 homolog. rdh54+ shares sequence and functional homology to budding yeast RDH54/TID1. Rdh54p is present during meiosis with appropriate timing for a meiotic recombination factor. It interacts with Rhp51 and the meiotic Rhp51 homolog Dmc1 in yeast two-hybrid assays. Deletion of rdh54+ has no effect on DNA damage repair during the haploid vegetative cell cycle. In meiosis, however, rdh54Delta shows decreased spore viability and homologous recombination with a concomitant increase in sister chromatid exchange. The rdh54Delta single mutant repairs meiotic breaks with similar timing to wild type, suggesting redundancy of meiotic recombination factors. Consistent with this, the rdh54Delta rhp54Delta double mutant fails to repair meiotic double strand breaks. Live cell analysis shows that rdh54Delta rhp54Delta asci do not arrest, but undergo both meiotic divisions with near normal timing, suggesting that failure to repair double strand breaks in S. pombe meiosis does not result in checkpoint arrest. PMID- 14551248 TI - Importin beta negatively regulates nuclear membrane fusion and nuclear pore complex assembly. AB - Assembly of a eukaryotic nucleus involves three distinct events: membrane recruitment, fusion to form a double nuclear membrane, and nuclear pore complex (NPC) assembly. We report that importin beta negatively regulates two of these events, membrane fusion and NPC assembly. When excess importin beta is added to a full Xenopus nuclear reconstitution reaction, vesicles are recruited to chromatin but their fusion is blocked. The importin beta down-regulation of membrane fusion is Ran-GTP reversible. Indeed, excess RanGTP (RanQ69L) alone stimulates excessive membrane fusion, leading to intranuclear membrane tubules and cytoplasmic annulate lamellae-like structures. We propose that a precise balance of importin beta to Ran is required to create a correct double nuclear membrane and simultaneously to repress undesirable fusion events. Interestingly, truncated importin beta 45-462 allows membrane fusion but produces nuclei lacking any NPCs. This reveals distinct importin beta-regulation of NPC assembly. Excess full length importin beta and beta 45-462 act similarly when added to prefused nuclear intermediates, i.e., both block NPC assembly. The importin beta NPC block, which maps downstream of GTPgammaS and BAPTA-sensitive steps in NPC assembly, is reversible by cytosol. Remarkably, it is not reversible by 25 microM RanGTP, a concentration that easily reverses fusion inhibition. This report, using a full reconstitution system and natural chromatin substrates, significantly expands the repertoire of importin beta. Its roles now encompass negative regulation of two of the major events of nuclear assembly: membrane fusion and NPC assembly. PMID- 14551250 TI - Cyclin aggregation and robustness of bio-switching. AB - During the cell cycle, Cdc2-cyclin B kinase abruptly becomes active and triggers the entry into mitosis/meiosis. Recently, it was found that inactive Cdc2-cyclin B is present in aggregates in immature starfish oocytes and becomes disaggregated at the time of its activation during maturation. We discuss a possible scenario in which aggregation of Cdc2-cyclin B dramatically enhances robustness of this activation. In this scenario, only inactive Cdc2-cyclin B can form aggregates, and the aggregates are in equilibrium with inactive Cdc2-cyclin B in solution. During maturation, the hormone-triggered inactivation of Myt1 depletes the soluble inactive Cdc2-cyclin B and the turnover leads to dissolution of the aggregates. This phase change, when coupled with the instability of the signaling network, provides a robust bio-switch. PMID- 14551249 TI - Localization and dynamics of Cdc2-cyclin B during meiotic reinitiation in starfish oocytes. AB - The Cdc2-cyclin B kinase has a central role in regulating the onset of M phase. In starfish oocytes, Cdc2-cyclin B begins to be activated approximately 10 min after application of maturation hormone, followed by accumulation in the nucleus then nuclear envelope breakdown. By immunofluorescence and by expressing a green fluorescent (GFP) chimera of cyclin B, we find that cyclin B is present in aggregates in the cytoplasm of immature oocytes. The aggregates disperse at approximately 10 min, suggesting that the dispersal is closely related to the activation of the kinase. Using cyclin B-GFP, the dispersion begins from the region containing the centrosomes. Extractability of Cdc2-cyclin B changes with similar kinetics during maturation. Active Cdc25 phosphatase released Cdc2-cyclin B from the detergent-insoluble fraction independently of its phosphatase activity. Live cell imaging also showed that Cdc2-cyclin B begins to accumulate in the nucleus before changes in nuclear pore permeability, consistent with Cdc2 cyclin B-induced disassembly of the pores. PMID- 14551251 TI - Targeted chemical disruption of clathrin function in living cells. AB - The accurate assignment of molecular roles in membrane traffic is frequently complicated by the lack of specific inhibitors that can work on rapid time scales. Such inhibition schemes would potentially avoid the complications arising from either compensatory gene expression or the complex downstream consequences of inhibition of an important protein over long periods (>12 h). Here, we developed a novel chemical tool to disrupt clathrin function in living cells. We engineered a cross-linkable form of clathrin by using an FK506-binding protein 12 (FKBP)-clathrin fusion protein that is specifically oligomerized upon addition of the cell-permeant cross-linker FK1012-A. This approach interrupts the normal assembly-disassembly cycle of clathrin lattices and results in a specific, rapid, and reversible approximately 70% inhibition of clathrin function. This approach should be applicable to a number of proteins that must go through an assembly disassembly cycle for normal function. PMID- 14551252 TI - Acroplaxome, an F-actin-keratin-containing plate, anchors the acrosome to the nucleus during shaping of the spermatid head. AB - Nuclear shaping is a critical event during sperm development as demonstrated by the incidence of male infertility associated with abnormal sperm ad shaping. Herein, we demonstrate that mouse and rat spermatids assemble in the subacrosomal space a cytoskeletal scaffold containing F-actin and Sak57, a keratin ortholog. The cytoskeletal plate, designated acroplaxome, anchors the developing acrosome to the nuclear envelope. The acroplaxome consists of a marginal ring containing keratin 5 10-nm-thick filaments and F-actin. The ring is closely associated with the leading edge of the acrosome and to the nuclear envelope during the elongation of the spermatid head. Anchorage of the acroplaxome to the gradually shaping nucleus is not disrupted by hypotonic treatment and brief Triton X-100 extraction. By examining spermiogenesis in the azh mutant mouse, characterized by abnormal spermatid/sperm head shaping, we have determined that a deformity of the spermatid nucleus is restricted to the acroplaxome region. These findings lead to the suggestion that the acroplaxome nucleates an F-actin-keratin-containing assembly with the purpose of stabilizing and anchoring the developing acrosome during spermatid nuclear elongation. The acroplaxome may also provide a mechanical planar scaffold modulating external clutching forces generated by a stack of Sertoli cell F-actin-containing hoops encircling the elongating spermatid nucleus. PMID- 14551253 TI - PIKfyve controls fluid phase endocytosis but not recycling/degradation of endocytosed receptors or sorting of procathepsin D by regulating multivesicular body morphogenesis. AB - The mammalian phosphatidylinositol (PtdIns) 5-P/PtdIns 3,5-P2-producing kinase PIKfyve has been implicated in maintaining endomembrane homeostasis in mammalian cells. To address the role of PIKfyve in trafficking processes, we examined the functioning of the biosynthetic, endocytic, and recycling pathways in stable human embryonic kidney 293 cell lines inducibly expressing the wild-type or kinase-defective dominant-negative form. PIKfyveWT or PIKfyveK1831E expression did not affect the processing and lysosomal targeting of newly synthesized procathepsin D. Likewise the rates of transferrin uptake/recycling or epidermal growth factor receptor degradation were not altered upon expression of either protein. In contrast, PIKfyveK1831E but not PIKfyveWT expression markedly impaired the late uptake of fluid phase marker horseradish peroxidase. Inspection of the organelle morphology by confocal microscopy with specific markers in COS cells transiently expressing PIKfyveK1831E showed the Golgi apparatus, end lysosomes, and the recycling compartment indistinguishable from nontransfected cells, despite the dramatic PIKfyveK1831E-induced endomembrane vacuolation. In contrast, we observed a striking effect on the late endocytic compartment, marked by disruption of the dextran-labeled perinuclear endosomal compartment and formation of dispersed enlarged vesicles. Electron microscopy identified the cytoplasmic vacuoles in the PIKfyveK1831E-expressing human embryonic kidney 293 cells as enlarged multivesicular body-like structures with substantially lower number of internal vesicles and membrane whorls. Together, these data indicate that PIKfyve selectively regulates the sorting and traffic of peripheral endosomes containing lysosomaly directed fluid phase cargo through controlling the morphogenesis and function of multivesicular bodies. PMID- 14551254 TI - Visualization of protein compartmentation within the plasma membrane of living yeast cells. AB - Different distribution patterns of the arginine/H+ symporter Can1p, the H+ plasma membrane ATPase Pma1p, and the hexose transport facilitator Hxt1p within the plasma membrane of living Saccharomyces cerevisiae cells were visualized using fluorescence protein tagging of these proteins. Although Hxt1p-GFP was evenly distributed through the whole cell surface, Can1p-GFP and Pma1p-GFP were confined to characteristic subregions in the plasma membrane. Pma1p is a well-documented raft protein. Evidence is presented that Can1p, but not Hxt1p, is exclusively associated with lipid rafts, too. Double labeling experiments with Can1p-GFP- and Pma1p-RFP-containing cells demonstrate that these proteins occupy two different nonoverlapping membrane microdomains. The size of Can1p-rich (Pma1p-poor) areas was estimated to 300 nm. These domains were shown to be stable in growing cells for >30 min. To our knowledge, this is the first observation of a cell polarization-independent lateral compartmentation in the plasma membrane of a living cell. PMID- 14551255 TI - Diacylglycerol kinase-zeta localization in skeletal muscle is regulated by phosphorylation and interaction with syntrophins. AB - Syntrophins are scaffolding proteins that link signaling molecules to dystrophin and the cytoskeleton. We previously reported that syntrophins interact with diacylglycerol kinase-zeta (DGK-zeta), which phosphorylates diacylglycerol to yield phosphatidic acid. Here, we show syntrophins and DGK-zeta form a complex in skeletal muscle whose translocation from the cytosol to the plasma membrane is regulated by protein kinase C-dependent phosphorylation of the DGK-zeta MARCKS domain. DGK-zeta mutants that do not bind syntrophins were mislocalized, and an activated mutant of this sort induced atypical changes in the actin cytoskeleton, indicating syntrophins are important for localizing DGK-zeta and regulating its activity. Consistent with a role in actin organization, DGK-zeta and syntrophins were colocalized with filamentous (F)-actin and Rac in lamellipodia and ruffles. Moreover, extracellular signal-related kinase-dependent phosphorylation of DGK zeta regulated its association with the cytoskeleton. In adult muscle, DGK-zeta was colocalized with syntrophins on the sarcolemma and was concentrated at neuromuscular junctions (NMJs), whereas in type IIB fibers it was found exclusively at NMJs. DGK-zeta was reduced at the sarcolemma of dystrophin deficient mdx mouse myofibers but was specifically retained at NMJs, indicating that dystrophin is important for the sarcolemmal but not synaptic localization of DGK-zeta. Together, our findings suggest syntrophins localize DGK-zeta signaling complexes at specialized domains of muscle cells, which may be critical for the proper control of lipid-signaling pathways regulating actin organization. In dystrophic muscle, mislocalized DGK-zeta may cause abnormal cytoskeletal changes that contribute to disease pathogenesis. PMID- 14551256 TI - Loss of SEC-23 in Caenorhabditis elegans causes defects in oogenesis, morphogenesis, and extracellular matrix secretion. AB - SEC-23 is a component of coat protein complex II (COPII)-coated vesicles involved in the endoplasmic reticulum-to-Golgi transport pathway of eukaryotes. During postembryonic life, Caenorhabditis elegans is surrounded by a collagenous exoskeleton termed the cuticle. From a screen for mutants defective in cuticle secretion, we identified and characterized a sec-23 mutant of C. elegans. By sequence homology, C. elegans has only the single sec-23 gene described herein. In addition to the cuticle secretion defect, mutants fail to complete embryonic morphogenesis. However, they progress through the earlier stages of embryogenesis, including gastrulation, and achieve substantial morphogenesis before death. We demonstrated a maternal component of SEC-23 function sufficient for progression through the earlier stages of embryogenesis and explaining the limited phenotype of the zygotic mutant. By RNA-mediated interference, we investigated the effects of perturbing COPII function during various postembryonic stages. During larval stages, major defects in cuticle synthesis and molting were observed. In the adult hermaphrodite, reduction of SEC-23 function by RNA-mediated interference caused a rapid onset of sterility, with defects in oogenesis including early maturation of the germline nuclei, probably a result of the observed loss of the GLP-1 receptor from the membrane surfaces adjacent to the developing germline nuclei. PMID- 14551257 TI - The role of the polo kinase Cdc5 in controlling Cdc14 localization. AB - In budding yeast, the protein phosphatase Cdc14 controls exit from mitosis. Its activity is regulated by a competitive inhibitor Cfi1/Net1, which binds to and sequesters Cdc14 in the nucleolus. During anaphase, Cdc14 is released from its inhibitor by the action of two regulatory networks. The Cdc Fourteen Early Anaphase Release (FEAR) network initiates Cdc14 release from Cfi1/Net1 during early anaphase, and the Mitotic Exit Network (MEN) promotes Cdc14 release during late anaphase. Here, we investigate the relationship among FEAR network components and propose an order in which they function to promote Cdc14 release from the nucleolus. Furthermore, we examine the role of the protein kinase Cdc5, which is a component of both the FEAR network and the MEN, in Cdc14 release from the nucleolus. We find that overexpression of CDC5 led to Cdc14 release from the nucleolus in S phase-arrested cells, which correlated with the appearance of phosphorylated forms of Cdc14 and Cfi1/Net1. Cdc5 promotes Cdc14 phosphorylation and, by stimulating the MEN, Cfi1/Net1 phosphorylation. Furthermore, we suggest that Cdc14 release from the nucleolus only occurs when Cdc14 and Cfi1/Net1 are both phosphorylated. PMID- 14551259 TI - Interaction with Tap42 is required for the essential function of Sit4 and type 2A phosphatases. AB - In Saccharomyces cerevisiae, Pph21 and Pph22 are the two catalytic subunits of type 2A phosphatase (PP2Ac), and Sit4 is a major form of 2A-like phosphatase. The function of these phosphatases requires their association with different regulatory subunits. In addition to the conventional regulatory subunits, namely, the A and B subunits for Pph21/22 and the Sap proteins for Sit4, these phosphatases have been found to associate with a protein termed Tap42. In this study, we demonstrated that Sit4 and PP2Ac interact with Tap42 via an N-terminal domain that is conserved in all type 2A and 2A-like phosphatases. We found that the Sit4 phosphatase in the sit4-102 strain contains a reverse-of-charge amino acid substitution within its Tap42 binding domain and is defective for formation of the Tap42-Sit4 complex. Our results suggest that the interaction with Tap42 is required for the activity as well as for the essential function of Sit4 and PP2Ac. In addition, we showed that Tap42 is able to interact with two other 2A like phosphatases, Pph3 and Ppg1. PMID- 14551258 TI - Urmylation: a ubiquitin-like pathway that functions during invasive growth and budding in yeast. AB - Ubiquitin is a small modifier protein that is conjugated to substrates to target them for degradation. Recently, a surprising number of ubiquitin-like proteins have been identified that also can be attached to proteins. Herein, we identify two molecular functions for the posttranslational protein modifier from Saccharomyces cerevisiae, Urm1p. Simultaneous loss of Urm1p and Cla4p, a p21 activated kinase that functions in budding, is lethal. This result suggests a role for the urmylation pathway in budding. Furthermore, loss of the urmylation pathway causes defects in invasive growth and confers sensitivity to rapamycin. Our results indicate that the sensitivity to rapamycin is due to a genetic interaction with the TOR pathway, which is important for regulation of cell growth in response to nutrients. We have found that Urm1p can be attached to a number of proteins. Loss of five genes that are also essential in a cla4Delta strain, NCS2, NCS6, ELP2, ELP6, and URE2, affect the level of at least one Urm1p conjugate. Moreover, these five genes have a role in invasive growth and display genetic interactions with the TOR pathway. In summary, our results suggest the urmylation pathway is involved in nutrient sensing and budding. PMID- 14551261 TI - Repression of p65 transcriptional activation by the glucocorticoid receptor in the absence of receptor-coactivator interactions. AB - Glucocorticoids are among the most potent antiinflammatory agents, acting through the glucocorticoid receptor (GR) to suppress gene expression of a variety of cytokines. This appears to be via transcriptional interference (or transrepression) of key regulatory factors such as nuclear factor-kappaB and activator protein 1. Ligand-bound GR can also activate gene transcription (transactivation) via direct binding to glucocorticoid response elements. Transactivation by GR is potentiated by accessory coactivators such as steroid receptor coactivator 1 and peroxisome proliferator-activated receptor gamma coactivator 1, whereas the role of these proteins in transrepression is unclear.Here, we show that GR can recruit several coactivator receptor interacting domains in a ligand-dependent manner. All interactions require the charge clamp defined by K579/E755, while a subset also requires a second charge clamp defined by R585/D590, within the GR ligand-binding domain. A point mutation, E755A, abolished all GR-receptor interacting domain interactions and led to a decrease in GR-mediated transactivation, but did not significantly affect GR-mediated transrepression of Gal4-p65 activity. Overexpression of a GR interacting coactivator peptide blocked transactivation but did not affect transrepression of p65 or TNFalpha-induced IL-6 promoter activity. Finally, the GR antagonist RU486 did not recruit coactivators to GR but maintained the ability to transrepress p65 activity. Our data suggest that different coactivators utilize distinct contact points to interact with GR. Although GR interactions with specific coactivators are critical for transactivation, they appear to be dispensable for at least certain aspects of GR-mediated transrepression of nuclear factor-kappaB. This is consistent with the notion that all GR- mediated repression is not intrinsically linked to activation and can be separated mechanistically. PMID- 14551260 TI - Significance of 14-3-3 self-dimerization for phosphorylation-dependent target binding. AB - 14-3-3 proteins via binding serine/threonine-phosphorylated proteins regulate diverse intracellular processes in all eukaryotic organisms. Here, we examine the role of 14-3-3 self-dimerization in target binding, and in the susceptibility of 14-3-3 to undergo phosphorylation. Using a phospho-specific antibody developed against a degenerated mode-1 14-3-3 binding motif (RSxpSxP), we demonstrate that most of the 14-3-3-associated proteins in COS-7 cells are phosphorylated on sites that react with this antibody. The binding of these phosphoproteins depends on 14 3-3 dimerization, inasmuch as proteins associated in vivo with a monomeric 14-3-3 form are not recognized by the phospho-specific antibody. The role of 14-3-3 dimerization in the phosphorylation-dependent target binding is further exemplified with two well-defined 14-3-3 targets, Raf and DAF-16. Raf and DAF-16 can bind both monomeric and dimeric 14-3-3; however, whereas phosphorylation of specific Raf and DAF-16 sites is required for binding to dimeric 14-3-3, binding to monomeric 14-3-3 forms is entirely independent of Raf and DAF-16 phosphorylation. We also find that dimerization diminishes 14-3-3 susceptibility to phosphorylation. These findings establish a significant role of 14-3-3 dimerization in its ability to bind targets in a phosphorylation-dependent manner and point to a mechanism in which 14-3-3 phosphorylation and dimerization counterregulate each other. PMID- 14551262 TI - Glucocorticoids control beta-catenin protein expression and localization through distinct pathways that can be uncoupled by disruption of signaling events required for tight junction formation in rat mammary epithelial tumor cells. AB - In Con8 rat mammary epithelial tumor cells, the synthetic glucocorticoid dexamethasone stimulates the remodeling of tight junctions and adherens junctions before formation of highly sealed tight junctions. In this study, the expression and localization of key components of the apical junction were examined as potential targets of glucocorticoid signaling. Western blot and RT-PCR demonstrated that dexamethasone up-regulated beta-catenin protein and transcript expression and nearly ablated beta-catenin phosphorylation under conditions that led to a significant increase in monolayer transepithelial resistance. Indirect immunofluorescence revealed that dexamethasone treatment also caused beta-catenin to localize predominantly at the cell membrane rather than the nucleus. The glucocorticoid regulation of beta-catenin expression and localization was not a consequence of dexamethasone inhibition of cell growth, because both responses were unaltered in the presence of hydroxyurea. The steroid induction of beta catenin expression and localization can be uncoupled by altering the function of signaling pathways needed for tight junction formation. Expression of dominant negative RasN17 abolished dexamethasone up-regulation of beta-catenin protein expression without affecting its localization at the membrane. In contrast, exogenous treatment or constitutive production of TGFalpha abolished the dexamethasone-induced alteration of beta-catenin localization without affecting the dexamethasone stimulation of beta-catenin expression. Taken together, our results demonstrate that glucocorticoids control beta-catenin at two distinct levels of cellular regulation that differ in their cell signaling requirements for the glucocorticoid regulation of mammary epithelial junctional dynamics. PMID- 14551263 TI - Activins are critical modulators of growth and survival. AB - Activins betaA and betaB (encoded by Inhba and Inhbb genes, respectively) are related members of the TGF-beta superfamily. Previously, we generated mice with an Inhba knock-in allele (InhbaBK) that directs the expression of activin betaB protein in the spatiotemporal pattern of activin betaA. These mice were small and had shortened life spans, both influenced by the dose of the hypomorphic InhbaBK allele. To understand the mechanism(s) underlying these abnormalities, we now examine growth plates, liver, and kidney and analyze IGF-I, GH, and major urinary proteins. Our studies show that activins modulate the biological effects of IGF-I without substantial effects on GH, and that activin signaling deficiency also has modest effects on hepatic and renal function. To assess the relative influences of activin betaA and activin betaB, we produced mice that express activin betaB from the InhbaBK allele, and not from its endogenous Inhbb locus. InhbaBK/BK, Inhbb-/- mice have failure of eyelid fusion at birth and demonstrate more severe effects on somatic growth and survival than either of the corresponding single homozygous mutants, showing that somatic growth and life span are supported by both activins betaA and betaB, although activin betaA plays a more substantial role. PMID- 14551264 TI - Novel activation step required for transcriptional competence of progesterone receptor on chromatin templates. AB - To elucidate the earliest molecular steps in the activation of transcription by the progesterone receptor (PR), we investigated its activity in a cell-free transcription system utilizing chromatin templates. PR prepared as a ligand-free, recombinant protein failed to induce transcription on chromatin templates. However, transcriptional competence could be restored by coincubation with rabbit reticulocyte lysate (RRL). The interaction of PR with chaperones results in a receptor conformation competent to bind ligand and RRL contains abundant chaperone-mediated protein folding activity. Blocking this activity with the specific inhibitor geldanamycin inhibited receptor-dependent transcriptional activity. However, recombinant chaperones could not replace RRL in the restoration of transcriptional activity on chromatin templates, suggesting the presence of an additional activity in the lysate. Under chromatin assembly conditions, PR could bind naked DNA and RRL did not increase that binding. In contrast, PR bound to a chromatin template only poorly. Interestingly, RRL stimulated sequence-specific binding by PR to target sites in chromatin and the concomitant recruitment of the steroid receptor coactivator 1 to the promoter. Thus, our results indicate that a novel protein-mediated activity in RRL is involved in an additional, heretofore unrecognized, activation step required for PR to become transcriptionally competent on chromatin templates. PMID- 14551265 TI - Hoxa11 regulates stromal cell death and proliferation during neonatal uterine development. AB - Increasing evidence indicates that the Hoxa11 gene plays a critical role in the proper development of the uterus. In this report, we describe potential altered cellular processes in the developing uterus of Hoxa11 mutants. Histologic analysis demonstrates normal uterine morphology in Hoxa11 mutants as compared with controls at the newborn stage and d 7 after birth. Stromal tissue was moderately reduced in the Hoxa11 mutant uterus by d 14 after birth and was absent by d 21 after birth. There is decreased cellular proliferation in the Hoxa11 mutant uterus both at 7 and 14 d after birth. Terminal deoxyribonucleotide transferase-mediated deoxyuridine triphosphate nick-end labeling analysis demonstrates that apoptosis was markedly increased in the Hoxa11 mutant uterus at d 14 after birth. p27 is decreased in the Hoxa11 mutant as evidenced by real-time PCR. Epidermal growth factor receptor expression is dramatically decreased as evidenced by both real-time PCR and immunohistochemistry results. These findings suggest that Hoxa11 is required for proper cellular proliferation and apoptotic responses in the developing neonatal uterus and that the regulation of epidermal growth factor receptor is critical to these processes. PMID- 14551266 TI - Recovery after uterine artery embolization: understanding and managing short-term outcomes. PMID- 14551267 TI - Digital image processing: a primer for JVIR authors and readers: part 1: the fundamentals. AB - Online submission of manuscripts will be mandatory for most journals in the near future. To prepare authors for this requirement and to acquaint readers with this new development, herein the basics of digital image processing are described. From the fundamentals of digital image architecture, through acquisition, editing, and storage of digital images, the steps necessary to prepare an image for online submission are reviewed. In this article, the first of a three-part series, the structure of the digital image is described. In subsequent articles, the acquisition and editing of digital images will be reviewed. PMID- 14551269 TI - Duplex ultrasound evaluation of lower extremity venous insufficiency. AB - Physicians unfamiliar with venous insufficiency, particularly disorders of the superficial venous system, often underestimate the complexity of the problem and the importance of proper evaluation before initiating treatment. In addition to a directed history evaluation and physical examination, additional evaluation with use of a variety of noninvasive diagnostic instruments, including duplex ultrasound, may be necessary when determining the cause, severity, and best treatment options available for a particular patient. After such evaluation, the treating physician should have a precise map of the patient's pathways of venous insufficiency, including sources of reflux (eg, saphenofemoral junction, saphenopopliteal junction, perforators), tributaries, vein size, and vein morphology. PMID- 14551270 TI - Tolerance, hospital stay, and recovery after uterine artery embolization for fibroids: the Ontario Uterine Fibroid Embolization Trial. AB - PURPOSE: Uterine artery embolization (UAE) is gaining popularity as an alternative to hysterectomy for the treatment of fibroids. Although minimally invasive treatments such as UAE offer the potential of fewer complications, shorter hospital stay, and quicker recovery than surgery, there have been few published data on tolerance and recovery in patients undergoing UAE. MATERIALS AND METHODS: This was a multicenter prospective single-arm clinical treatment trial involving the practices of 11 interventional radiologists in eight Ontario university-affiliated and community hospitals. Between November 1998 and November 2000, 555 women underwent UAE for symptomatic uterine fibroids. Follow-up included ultrasound examinations and telephone interviews. UAE was performed under conscious sedation. Polyvinyl alcohol particles (355-500 micro m) were the primary embolic agent, and the procedural endpoint involved stasis in the uterine arteries. Pain protocols included antiinflammatory medications and narcotics and a planned overnight hospital admission. Tolerance and recovery were measured by patient-reported pain intensity (10-point numeric rating and five-point descriptor scale), hospital length of stay (LOS), and time until return to work. RESULTS: Intraprocedural pain was reported by 30% of patients and postprocedural pain was reported by 92% of patients (mean pain rating +/- SD, 7.0 +/- 2.47). The mean hospital LOS was 1.3 nights. Postprocedural pain was the most common indication for an LOS greater than 1 night (18%) or 2 nights (5%). Return visits to the hospital (10%) and readmissions (3%) were primarily for pain. The overall postprocedural complication rate was 8.0% (95% CI: 5.9%-10.6%). Of the 44 complications, 32 (73%) were pain-related. The mean recovery time after UAE was 13.1 days (median, 10.0 d). CONCLUSION: The majority of patients had a 1-night LOS after UAE and recovered within 2 weeks. Postprocedural pain varied considerably and was the major indication for extended hospital stay and recovery. PMID- 14551271 TI - Hepatic perfusion as a predictor of mortality after transjugular intrahepatic portosystemic shunt creation in patients with refractory ascites. AB - PURPOSE: To determine whether hepatic perfusion patterns predict mortality after transjugular intrahepatic portosystemic shunt (TIPS) creation in patients with severe ascites. MATERIALS AND METHODS: This retrospective study included 22 patients who had enhanced cine magnetic resonance (MR) imaging performed immediately before TIPS creation in the angled coronal plane including the left kidney, liver, and main portal vein. Regions of interest were centered over the liver and kidney, and perfusion curves were generated and reviewed before the standard TIPS procedure was performed. Four patients did not undergo TIPS creation as a result of very poor hepatic perfusion by MR. All patients were followed clinically and by ultrasound surveillance of their shunt. RESULTS: Eleven patients died within 6 months, including all four patients who did not have a TIPS because of MR evidence of poor hepatic perfusion. Of these 11 patients, eight (73%) had unfavorable liver flow consisting of diminished enhancement compared to the kidney and early peak enhancement of less than 50 seconds. The surviving patients all showed a delayed peak enhancement of greater than 50 seconds. CONCLUSIONS: In patients undergoing TIPS creation for refractory ascites, blunted arterial-type hepatic enhancement is a poor prognostic sign. Cine MR imaging with evaluation of hepatic perfusion can be performed and reviewed before the TIPS procedure. Alternative techniques for ascites reduction may be preferred for patients with unfavorable hepatic perfusion. PMID- 14551272 TI - The Gunther Tulip retrievable filter: prolonged temporary filtration by repositioning within the inferior vena cava. AB - PURPOSE: To report experience with the retrievable Gunther Tulip filter (GTF) as a means of temporary caval filtration for the prevention of pulmonary embolism (PE) with use of a technique that prolongs filter dwell time beyond 14 days. MATERIALS AND METHODS: Eighty-eight GTFs were implanted in 87 patients. The GTFs were placed with the intention of retrieval in all patients within 14 days after initial implantation. In 23 of the 87 patients (26%), there was a need to prolong temporary caval filtration beyond the recommended period of 14 days. This was successfully achieved with use of percutaneous techniques from the right internal jugular vein whereby the filter was repositioned to a different location within the inferior vena cava (IVC) before definitive device removal. RESULTS: Of 88 GTFs implanted in 87 patients, 70 were successfully retrieved and 18 were left in place permanently. Forty-seven filters in 46 patients were removed after initial implantation with no need for percutaneous repositioning within the IVC to prolong dwell time (mean dwell time, 13 days). In the 23 patients who required repositioning of 23 GTFs within the IVC to prolong temporary caval filtration, the mean dwell time was 34.8 days; the mean number of repositioning procedures was 1.5, the mean time between repositioning procedures was 13.8 days, and the mean fluoroscopy time was 4.4 minutes in patients in whom filter retrieval was attempted. One patient underwent placement and subsequent removal of the GTF twice for perioperative prophylaxis against PE on two separate occasions. No filters were misplaced in an unintended location or tilted (>15 degrees ) in relation to the main caval axis after deployment. In one patient, a GTF became permanently fixed in the IVC 16 days after initial implantation and could not be removed percutaneously. Nine patients had mild or moderate-sized cervical hematomas. One patient had recurrent asymptomatic PE 2 months after filter insertion. CONCLUSION: Dwell times of 14 days can be achieved in most patients before device removal. Prolongation of the dwell time beyond 14 days can be safely and easily achieved by performing percutaneous repositioning of the device within the IVC via a jugular approach. PMID- 14551273 TI - Influence of large peritumoral vessels on outcome of radiofrequency ablation of liver tumors. AB - PURPOSE: The effect of large vessels (>/=3 mm) contiguous to hepatic tumors was evaluated with respect to clinical tumor recurrence rates after radiofrequency (RF) ablation. MATERIALS AND METHODS: The first 105 malignant liver tumors treated by RF ablation therapy at our institution with pathologic analysis or a minimum of 6 months of clinical follow-up were reviewed. The original pretreatment imaging studies were reviewed by a radiologist who was blinded to the cases, and, based on lesion contiguity to vessels of at least 3 mm, the lesions were categorized as perivascular or nonperivascular. Treatment outcomes with respect to local tumor recurrence between these two groups were then compared. Logistic regression analysis was performed to take into account other variables and to determine whether this categorization was an independent predictor of treatment outcome. RESULTS: There were 74 nonperivascular tumors and 31 perivascular tumors. Mean tumor size was 2.4 cm and mean follow-up was 11.3 months. Residual or locally recurrent tumors were documented in 20 of 105 cases (19%). In the nonperivascular group, five of 74 (7%) had either incompletely treated tumor (manifested within 6 months) or local recurrence beyond 6 months. In the perivascular group, 15 of 31 (48%) had incompletely treated or locally recurrent tumor (P <.001). Subanalysis of lesion size (61 tumors 4 cm), tumor type (40 hepatocellular carcinomas, 48 colorectal metastases, and 17 other metastases), access (53 intraoperative, 52 percutaneous), and RF device (45 Radiotherapeutics electrodes, 18 Rita electrodes, and 42 Radionics electrodes) showed similar results. Multivariate logistic regression analysis showed that presence or absence of a large peritumoral vessel is an independent, and the dominant, predictor of treatment outcome. CONCLUSION: The presence of vessels at least 3 mm in size contiguous to hepatic tumors is a strong independent predictor of incomplete tumor destruction by RF ablation. Modified ablation strategies should be considered to improve destruction of these tumors. PMID- 14551274 TI - Feasibility of magnetic resonance imaging-guided focused ultrasound surgery as an adjunct to tamoxifen therapy in high-risk surgical patients with breast carcinoma. AB - PURPOSE: To evaluate the feasibility of treating breast neoplasms with use of magnetic resonance (MR) imaging-guided focused ultrasound (US) surgery. MATERIALS AND METHODS: Twenty-four female patients, each with a single biopsy-proven breast carcinoma, who were considered to be at increased surgical risk or who had refused surgery underwent MR imaging-guided focused US surgery as an adjunct to their chemotherapeutic regimen of tamoxifen. Follow-up included routine studies to rule out metastatic disease and MR studies with and without contrast material infusion in the treated breast (10 days and 1, 3, and 6 months after the treatment session). Percutaneous biopsy was performed after 6-month follow-up, and if residual tumor was present, a second MR imaging-guided focused US surgery treatment session was performed, followed by repeat biopsy 1 month later. RESULTS: Twenty-three of 24 patients completed the protocol, with only one minor complication associated with the treatment sessions (second-degree skin burn resolved with local treatment). Follow-up MR studies demonstrated a varying hypointense treatment margin (range, 1-11 mm), which represents destruction of tissue beyond the visible tumor. Absence of enhancement may be an indicator of tumor destruction (18 of 19 patients with negative biopsy results) whereas persistent enhancement suggested tumor residue (three of five patients with residual tumor). Overall, 19 of 24 patients (79%) had negative biopsy results after one or two treatment sessions. CONCLUSION: MR imaging-guided focused US surgery of breast tumors is a safe, repeatable, and promising method of focal tumor destruction. PMID- 14551275 TI - De novo placement of button gastrostomy catheters in an adult population: experience in 53 patients. AB - PURPOSE: To investigate the feasibility of primary button gastrostomy insertion with the aid of T-fastener gastropexy. MATERIALS AND METHODS: Fifty-three consecutive patients (33 men, 20 women; mean age, 63.4 years) referred for percutaneous radiologic gastrostomy (PRG) underwent primary button gastrostomy insertion over an 18-month period in two centers. Nine of the patients (17%) were referred after failed endoscopic gastrostomy and 44 (83%) were primarily referred for PRG. Indications for gastrostomy included esophageal/head and neck malignancy (n = 33) and neurologic disorders (n = 20). Gastropexy with three or four T fasteners was performed in all patients and angioplasty balloon catheters (6 mm x 40 mm) were used to measure tract length and dilate the tract. An 18-F dilator was used for final tract dilation. Button gastrostomy catheters with retention balloons were inserted in all patients. Patient follow-up was performed by the department of dietetics, which contacted patients on a weekly basis. RESULTS: Primary button gastrostomy insertion was successful in 52 of 53 patients (98%). The mean gastrostomy button catheter survival was 13.3 weeks (range, 1-28 weeks). No episodes of button occlusion occurred. Since the beginning of this study, 33 patients (63%) have had their gastrostomy buttons replaced. The reasons for button replacement include burst retention balloons (n = 27; 52%), dislodgment of the catheter (n = 4; 8%), and continuing pain/discomfort at the gastrostomy site (n = 2; 4%). CONCLUSION: Button-type gastrostomy catheters can be placed de novo by interventional radiologists without the need for a mature tract, provided a T fastener gastropexy is used. The balloon retention button devices are not compromised by occlusion but do tend to become dislodged. PMID- 14551277 TI - Flared polyurethane-covered self-expandable nitinol stent for malignant biliary obstruction. AB - PURPOSE: To determine the technical efficacy and safety of a flared polyurethane covered self-expandable nitinol stent in the management of malignant biliary obstruction and to evaluate its clinical efficacy by estimating stent patency and patient survival rates. MATERIALS AND METHODS: Thirteen patients with common bile duct strictures (nonhilar) caused by malignant disease were treated by placement of 13 nitinol stents. The stents used include a flared section in the proximal portion (12 mm in diameter and 10 mm in length) and a section in the remnant portion that is fully covered with high-elasticity polyurethane, with an unconstrained diameter of 10 mm and a total length of 50-80 mm. Patient survival and stent patency rates were calculated with use of Kaplan-Meier survival analysis. The follow-up bilirubin and serum amylase and lipase levels were calculated, and the differences in means were evaluated with use of a Wilcoxon signed-rank test. The average follow-up duration was 22.9 weeks (range, 8-56 weeks). RESULTS: Placement was successful in all cases. The 30-day mortality rate was 0%. The survival rates were 38% and 24% at 20 and 50 weeks, respectively. Seventy-seven percent of study patients had adequate palliative drainage during their the remainder of their lives. The stent patency rates were 71% and 48% at 20 and 50 weeks, respectively. Three patients (23%) presented with stent occlusion requiring repeat intervention. There were no procedure-related complications such as proximal or distal migration. No complications occurred other than stent occlusion. One patient's stent was removed under endoscopic guidance 15 weeks after its insertion. Bilirubin levels had significantly decreased 1 week after stent insertion (P <.001). CONCLUSION: Preliminary results suggest that placement of a flared polyurethane-covered self-expandable nitinol stent is feasible and effective in achieving biliary drainage. The stents do not migrate, but there is tumor ingrowth into the flared portion of the stent. Treatment of a larger group of patients will be mandatory to validate these long term results. PMID- 14551278 TI - A canine model for studying endoleak after endovascular aneurysm repair. AB - PURPOSE: The aim of this study was to create an animal model of endoleak after stent-graft placement for abdominal aortic aneurysm (AAA) in which a large aneurysmal sac would be preserved for the testing of techniques for its percutaneous occlusion. MATERIALS AND METHODS: Infrarenal AAAs were created in nine dogs by anastomosis of an isolated segment of the inferior vena cava to the right side of the abdominal aorta in combination with a large anterior patch from the external jugular vein. One hour later, animals underwent percutaneous implantation of polytetrafluoroethylene-covered Z stent endografts with three 3 mm-diameter holes through the fabric. Aortograms were obtained before and after surgery, after endograft placement, and at the time of animal sacrifice at 1 week or 1, 2, 3, or 6 months. Pressures within the aorta and the aneurysm sac were recorded before animal sacrifice. Gross and histologic evaluations of the specimens were then carried out. RESULTS: Immediately after endograft placement, all nine animals had artificial type III endoleaks with angiographic filling of lumbar arteries and veins. One animal died of surgical complications within 2 days of surgery and is not included in our data analysis. One aneurysm ruptured at 1 week. At completion of the study, six endografts were patent and two were occluded. The aneurysm sac had enlarged by approximately 50% in seven animals. At follow-up, type I endoleak was present in three animals, type II endoleak was present in three, and the artificial type III endoleak was present in all six animals with patent endografts. The pressure differential between aorta and aneurysm sac was 36 mm Hg, with a mean aortic pressure of 87 mm Hg +/- 13.3 and a mean aneurysmal sac pressure of 51 mm Hg +/- 28.1. The aneurysmal sac exhibited early thrombus formation at 1 week, which progressed to complete thrombosis in 1 6 months. CONCLUSIONS: The model is technically feasible but would be useful in testing occlusive techniques for residual aneurysm sacs only in the acute phase after endograft placement. It would be not reliable for chronic evaluation because of rapidly progressive thrombosis in most aneurysm sacs and occasional complete thrombosis of the AAA and endograft. PMID- 14551279 TI - Relative rates of blood flow reduction during transcatheter arterial embolization with tris-acryl gelatin microspheres or polyvinyl alcohol: quantitative comparison in a swine model. AB - PURPOSE: To determine whether two commonly used embolic agents have differing rates of blood flow reduction during transcatheter embolization of the renal arteries in an animal model. MATERIALS AND METHODS: The renal arteries of 10 pigs were embolized with either polyvinyl alcohol (300-500 or 500-700- micro m) or tris-acryl gelatin microspheres (300-500 or 700-900- micro m). Equivalent unit doses of each agent were suspended in 40 mL of fluid and injected in 1-mL aliquots for 20 mL and then in 5-mL aliquots for 20 mL. Blood flow was measured after each aliquot with an intraarterial Doppler flow wire placed through the embolization catheter. RESULTS: Renal arterial blood flow was most rapidly and reliably decreased by 300-500- and 700-900- micro m microspheres, both of which had achieved >90% reduction from baseline flow after the injection of 6 mL of suspension. An equivalent reduction in flow required 25 mL of 300-500- micro m PVA suspension and 30 mL of 500-700- micro m PVA suspension. The reduction in blood flow with microspheres was significantly greater (P <.05) than that with PVA between 5 and 19 mL of suspension delivered. Differences between larger and smaller particle sizes of the same agent were not statistically significant. CONCLUSIONS: Tris-acryl gelatin microspheres reduced renal blood flow more quickly and reliably than did PVA. The type of agent used in embolization had a greater impact on the rate of flow reduction than did particle size in the range of sizes tested. PMID- 14551280 TI - In vivo intravascular MR imaging: transvenous technique for arterial wall imaging. AB - PURPOSE: To determine, in vivo, the potential for transvenous magnetic resonance (MR) imaging of the arterial wall and to assess appropriate MR pulse sequences for this method. MATERIALS AND METHODS: MR imaging was performed on 19 vessels (right renal artery, N = 9; left renal artery N = 2; external iliac artery, N = 4; abdominal aorta, N = 4) in nine swine. The animals were either low-density lipoprotein receptor knockout (N = 5) or Yucatan mini-pigs fed an atherogenic diet for 6 to 11 weeks (N = 4). The intravascular MR coil/guide wire (IVMRG) (Surgi-Vision, Gaithersburg, MD) was introduced via the external iliac vein into the inferior vena cava (IVC). The following electrocardiograph-gated MR pulse sequences were obtained: T1-weighted precontrast with and without fat saturation and T1-weighted postcontrast with fat saturation. Two observers scored wall signal and conspicuity and classified the vessel as normal, abnormal, or stented. Images were compared with histopathologic findings. RESULTS: The T1-weighted precontrast without fat saturation, T1-weighted precontrast with fat saturation, and T1-weighted postcontrast images correlated with histopathologic findings in 12 of 15 vessels, eight of 10 vessels, and 14 of 16 vessels, respectively. Abnormal histopathologic findings included: arterial wall thickening (N = 3), arterial dissection (N = 2), focal fibrous plaque (N = 2), adherent thrombus (N = 1). The T1-weighted postcontrast images were not compromised by artifacts and had the highest score for vessel wall signal and conspicuity. T1-weighted precontrast images were compromised by chemical shift artifact and poor blood suppression. Negligible artifacts were created by the platinum stent. CONCLUSION: The T1 weighted fat saturated postcontrast pulse sequence was superior to other sequences for transvenous MR imaging of the arterial wall. PMID- 14551281 TI - Ovarian protection by occlusion of uteroovarian collateral vessels before uterine fibroid embolization. AB - In an attempt to decrease the incidence of premature ovarian failure, three patients with prominent ovarian collateral vessels from the uterine artery (UA) underwent collateral vessel embolization before uterine fibroid embolization. UA anatomy and collateral pathways are reviewed. PMID- 14551282 TI - Coil embolization of a tuboovarian anastomosis before uterine artery embolization to prevent nontarget particle embolization of the ovary. AB - Uterine artery embolization (UAE) is being used more frequently as a primary treatment for uterine leiomyoma. Performing UAE in women who desire future fertility is controversial because of the risks of premature menopause and the undetermined effects on pregnancy. The etiology of ovarian failure after UAE is not yet clearly defined, but one of the leading possibilities is nontarget embolization of the ovaries. In this case report, the authors describe a technique of selective coil embolization of a uterine artery-to-ovarian artery communication before UAE performed specifically to protect the ovary from nontarget embolization. PMID- 14551283 TI - Transatrial stent placement for treatment of inferior vena cava obstruction secondary to extension of intracardiac tumor thrombus from hepatocellular carcinoma. AB - Two cases of superior vena cava-to-inferior vena cava (IVC) transatrial stent placement to palliate obstruction of the IVC secondary to the intracardiac extension of hepatocellular carcinoma are reported. Both patients presented with debilitating edema of the trunk and lower extremities and varying degrees of hepatic venous obstruction resulting in Budd-Chiari syndrome. One patient required the adjunctive creation of a percutaneous portocaval shunt and the second patient responded to transatrial stent placement alone. Both patients' functional status improved and edema markedly decreased after endovascular therapy. PMID- 14551284 TI - Radiofrequency ablation in a previously irradiated liver. AB - A patient with renal cell carcinoma underwent external-beam radiation therapy (XRT) to treat a painful chest-wall metastasis. One month later, she underwent radiofrequency (RF) ablation of two metastatic deposits within the liver; one of the target lesions was in the recent irradiation zone and the other was outside of the radiation field. RF ablation within the irradiated liver produced a slightly larger ablation zone with prominent needle tract scarring, and required less energy input than treatment in the unirradiated liver. RF ablation and XRT may interact, possibly producing a synergistic effect. Further study of the potentially adjunctive relationship between these two modalities is warranted. PMID- 14551285 TI - Complex endovascular repair of an aortobiiliac aneurysm with use of a combination of commercially available devices. PMID- 14551286 TI - Intra-accumbal injection of CART (cocaine-amphetamine regulated transcript) peptide reduces cocaine-induced locomotor activity. AB - Evidence suggests that CART (cocaine-amphetamine regulated transcript) peptides are mediators or modulators of the actions of psychostimulant drugs. In this study, the effects of intra-accumbal injections of rat long form (rl) CART 55-102 were examined. Injection of the peptide alone had no effect, but pretreatment with the peptide blunted or reduced the locomotor-inducing effects of cocaine after an i.p. injection. This effect was dose related and time limited, as expected. rlCART 1-27, a CART peptide fragment not active in other studies, was without effect on cocaine-induced locomotor activity. Because the actions of cocaine involve dopamine, the effect of rlCART 55-102 on dopamine-induced locomotor activity was examined. Intraaccumbal injection of dopamine produced a dose-related and time-limited increase in locomotor activity, as expected. Coinjection of rlCART 55-102 with dopamine blunted the effect. In summary, these data suggest that CART peptides in the nucleus accumbens would tend to oppose the actions of cocaine. PMID- 14551287 TI - Hepatic CYP2B6 expression: gender and ethnic differences and relationship to CYP2B6 genotype and CAR (constitutive androstane receptor) expression. AB - CYP2B6 metabolizes many drugs, and its expression varies greatly. CYP2B6 genotype phenotype associations were determined using human livers that were biochemically phenotyped for CYP2B6 (mRNA, protein, and CYP2B6 activity), and genotyped for CYP2B6 coding and 5'-flanking regions. CYP2B6 expression differed significantly between sexes. Females had higher amounts of CYP2B6 mRNA (3.9-fold, P < 0.001), protein (1.7-fold, P < 0.009), and activity (1.6-fold, P < 0.05) than did male subjects. Furthermore, 7.1% of females and 20% of males were poor CYP2B6 metabolizers. Striking differences among different ethnic groups were observed: CYP2B6 activity was 3.6- and 5.0-fold higher in Hispanic females than in Caucasian (P < 0.022) or African-American females (P < 0.038). Ten single nucleotide polymorphisms (SNPs) in the CYP2B6 promoter and seven in the coding region were found, including a newly identified 13072A>G substitution that resulted in an Lys139Glu change. Many CYP2B6 splice variants (SV) were observed, and the most common variant lacked exons 4 to 6. A nonsynonymous SNP in exon 4 (15631G>T), which disrupted an exonic splicing enhancer, and a SNP 15582C>T in an intron-3 branch site were correlated with this SV. The extent to which CYP2B6 variation was a predictor of CYP2B6 activity varied according to sex and ethnicity. The 1459C>T SNP, which resulted in the Arg487Cys substitution, was associated with the lowest level of CYP2B6 activity in livers of females. The intron-3 15582C>T SNP (in significant linkage disequilibrium with a SNP in a putative hepatic nuclear factor 4 (HNF4) binding site) was correlated with lower CYP2B6 expression in females. In conclusion, we found several common SNPs that are associated with polymorphic CYP2B6 expression. PMID- 14551288 TI - Evidence for a selective role of the delta-opioid agonist [8R (4bS*,8aalpha,8abeta, 12bbeta)]7,10-Dimethyl-1-methoxy-11-(2 methylpropyl)oxycarbonyl 5,6,7,8,12,12b-hexahydro-(9H)-4,8-methanobenzofuro[3,2 e]pyrrolo[2,3-g]isoquinoline hydrochloride (SB-235863) in blocking hyperalgesia associated with inflammatory and neuropathic pain responses. AB - The specific involvement of the delta-opioid receptor in the control of nociception was explored by investigating the pharmacological activity in vivo of a selective, orally active, and centrally penetrant delta-opioid agonist. [8R (4bS*,8aalpha,8abeta,12bbeta)]7,10-dimethyl-1-methoxy-11-(2 methylpropyl)oxycarbonyl 5,6,7,8,12,12b-hexahydro-(9H)-4,8-methanobenzofuro[3,2 e]pyrrolo[2,3-g]isoquinoline hydrochloride (SB-235863) is a new pyrrolomorphinan with high affinity (Ki = 4.81 +/- 0.39 nM) for the delta-opioid receptor, full agonist activity, and binding selectivity versus the mu- and kappa-opioid receptors of 189-fold and 52-fold, respectively. Perorally administered SB-236863 was inactive in the rat tail-flick and hot-plate tests of acute pain response, but potently reversed thermal hyperalgesia in rats resulting from a carrageenan induced inflammatory response. This activity could be blocked by the delta-opioid antagonist naltrindole (3 mg/kg s.c.), but selective mu- and kappa-opioid antagonists were ineffective. Naltrindole (1 microg i.c.v.) also blocked the activity of 10 mg/kg (p.o.) SB-235863, showing that the compound activates delta opioid receptor sites in the central nervous system. SB-235863 was additionally effective at reversing chronic hyperalgesia in the Seltzer rat model of partial sciatic nerve ligation after peroral administration. These data show that the delta-opioid receptor plays a selective role in regulating evoked and lasting changes in nociceptive pain signaling. Classical side effects of mu- and kappa opioid receptor activation (slowing of gastrointestinal transit and motor incoordination, respectively) were not observed after administration of 70 mg/kg (p.o.) SB-235863, nor was evoked seizure activity affected. These results suggest a selective and limited role of delta-opioid receptors in the modulation of nociception. PMID- 14551289 TI - Neuropeptide Y Y4 receptor homodimers dissociate upon agonist stimulation. AB - The pancreatic polypeptide-fold family of peptides consists of three 36-amino acid peptides, namely neuropeptide Y (NPY), peptide YY, and pancreatic polypeptide (PP). These peptides regulate important functions, including food intake, circadian rhythms, mood, blood pressure, intestinal secretion, and gut motility, through four receptors: Y1, Y2, Y4, and Y5. Additional receptor subtypes have been proposed based on pharmacology observed in native tissues. Recent studies with other G-protein-coupled receptors have shown that homo- and heterodimerization may be important in determining receptor function and pharmacology. In the present study, the recently cloned rhesus (rh) Y4 receptor was evaluated using radioligand binding, and the pharmacological profile was found to be very similar to the human Y4 receptor. To study homo- and heterodimerization involving the Y4 receptor using bioluminescence resonance energy transfer 2 (BRET(2)), the carboxy termini of the rhesus Y1, Y2, Y4, and Y5 receptors were fused to Renilla luciferase, and rhY4 was also fused to green fluorescent protein. Dimerization was also studied using Western blot analysis. Using both BRET(2) and Western analysis, we found that the rhY4 receptor is present at the cell surface as a homodimer. Furthermore, agonist stimulation using the Y4-selective agonists PP and 1229U91 can dissociate these dimers in a concentration-dependent manner. In contrast, rhY4 did not heterodimerize with other members of the NPY receptor family or with human opioid delta and mu receptors. Therefore, homodimerization is an important component in the regulation of the Y4 receptor. PMID- 14551290 TI - Antiallodynic effects of intrathecal orexins in a rat model of postoperative pain. AB - Orexin A and B (hypocretin 1 and 2) are the endogenous ligands of orexin receptors, a G-protein-coupled orphan receptor family containing orexin 1 (OX1) and orexin 2 (OX2) types. Orexin A induces analgesia in acute and inflammatory pain models. We further elucidated the possible antiallodynic effect of intrathecal orexins in a rat model of postoperative pain. Mechanical allodynia was induced by incising the rat hind paw and evaluated with the withdrawal threshold to von Frey filament stimulation. Intrathecal orexin A (0.03-1 nmol) and orexin B (0.1-3 nmol) dose dependently attenuated the incision-induced allodynia. Orexin A (ED50 = 0.06 nmol) is more potent than orexin B. The effects of orexin A and B were abolished by their respective antibodies, but not by naloxone, and were attenuated by suramin and strychnine, the P2X purinergic and glycine receptor antagonists, respectively. SB-334867, an OX1 receptor antagonist, at 30 nmol completely blocked the effect of orexin A but, even at 100 nmol, only partially antagonized the effect of orexin B. Orexin A antibody, SB 334867, suramin, strychnine, or naloxone enhanced the incision-induced allodynic response. It is concluded that intrathecal orexins reduce incision-induced allodynia through OX1 receptors. Glycine and P2X purinergic receptors, but not opioid receptors, might be involved in the antiallodynic effects of orexins. Endogenous orexin might be released after incision injury to activate the spinal OX1 receptors as an endogenous analgesic protector. PMID- 14551291 TI - Critical role of histamine H4 receptor in leukotriene B4 production and mast cell dependent neutrophil recruitment induced by zymosan in vivo. AB - The recently identified histamine receptor, H4, was shown to be primarily expressed on leukocytes and has been implicated in the activation of lymphocytes, eosinophils, and mast cells in vitro. Its function in vivo, however, has not yet been characterized. We present evidence for a critical role of H4 receptor in the mast cell-dependent recruitment of neutrophils. Mice injected with zymosan into the pleural cavity developed massive neutrophilia within hours after challenge. Neutrophilia was dose-dependently reduced when mice were pretreated with thioperamide, a known H(3/4) receptor antagonist, whereas H1 and H2 receptor antagonists lacked efficacy. Similarly, a 70 to 80% reduction in neutrophils in the pleural cavity compared with wild-type animals was noted in mice lacking mast cells (W/W(v) mice); mice deficient in MyD88 (MyD88(-/-)); a critical component of the signaling cascade of the major receptor for zymosan, toll-like receptor 2 (TLR2); or in mice pretreated with a functionally antagonistic anti-TLR2 antibody. The residual 20% neutrophil infiltration seen in mast cell-deficient and MyD88(-/-) mice was not further reduced by thioperamide. Neutrophilia was completely restored by transferring wild-type bone marrow-derived mast cells into MyD88(-/-) or W/W(v) mice. Interestingly, when neutrophilia was evoked by carrageenan injection, mast cell depletion and thioperamide had no effect. Various inflammatory mediators were detectable in the pleural cavity of zymosan challenged mice. Upon pretreatment with thioperamide, reduced levels of the neutrophil chemoattractant leukotriene B4 were observed, providing a mechanistic explanation for the prevention of neutrophilia by H4 receptor antagonism. PMID- 14551292 TI - Adverse prognostic effect of methylation in colorectal cancer is reversed by microsatellite instability. AB - PURPOSE: DNA methylation is an important biologic event in colorectal cancer and in some cases is associated with the development of microsatellite instability (MSI). In this study, we sought to determine the prognostic significance of DNA methylation, both in univariate analysis and in concert with other clinicopathologic factors known to influence outcome. PATIENTS AND METHODS: Fresh tissue (625 cancers) was obtained from 605 individuals (age range, 29 to 99 years) undergoing curative surgery for colorectal cancer at one institution during a period of 8 years. Clinicopathologic details were recorded for all tumors, including stage, grade, type, vascular space invasion, and clinical follow-up to 5 years. Microsatellite status was assessed using standard markers. Methylation of p16 and hMLH1 promoters was determined by methylation-specific polymerase chain reaction (PCR), whereas methylation at methylated-in-tumor loci (MINT)1, MINT2, MINT12, and MINT31 loci were assessed by bisulfite-PCR. RESULTS: Patients with microsatellite unstable tumors (12%) had better disease-specific survival than those with microsatellite stable (MSS) tumors (univariate analysis: hazard ratio [HR], 0.53; 95% CI, 0.27 to 1.0). Overall survival of individuals with MSS tumors was influenced by three independently significant factors: tumor stage (HR, 7.3; 95% CI, 5.1 to 10.4), heavy tumor methylation (HR, 2.1; 95% CI, 1.1 to 4.0), and vascular space invasion (HR, 1.9; 95% CI, 1.3 to 2.9). In MSS tumors, methylation at any single site was not independently predictive of survival. Neither methylation nor microsatellite status predicted a favorable response to chemotherapy. CONCLUSION: DNA methylation is associated with a worse outcome in colorectal cancer, but this adverse prognostic influence is lost in those methylated tumors showing MSI. The mechanisms of these events warrant additional investigation. PMID- 14551293 TI - Randomized trial of cytoreduction and hyperthermic intraperitoneal chemotherapy versus systemic chemotherapy and palliative surgery in patients with peritoneal carcinomatosis of colorectal cancer. AB - PURPOSE: To confirm the findings from uncontrolled studies that aggressive cytoreduction in combination with hyperthermic intraperitoneal chemotherapy (HIPEC) is superior to standard treatment in patients with peritoneal carcinomatosis of colorectal cancer origin. PATIENTS AND METHODS: Between February 1998 and August 2001, 105 patients were randomly assigned to receive either standard treatment consisting of systemic chemotherapy (fluorouracil leucovorin) with or without palliative surgery, or experimental therapy consisting of aggressive cytoreduction with HIPEC, followed by the same systemic chemotherapy regime. The primary end point was survival. RESULTS: After a median follow-up period of 21.6 months, the median survival was 12.6 months in the standard therapy arm and 22.3 months in the experimental therapy arm (log-rank test, P =.032). The treatment-related mortality in the aggressive therapy group was 8%. Most complications from HIPEC were related to bowel leakage. Subgroup analysis of the HIPEC group showed that patients with 0 to 5 of the 7 regions of the abdominal cavity involved by tumor at the time of the cytoreduction had a significantly better survival than patients with 6 or 7 affected regions (log rank test, P <.0001). If the cytoreduction was macroscopically complete (R-1), the median survival was also significantly better than in patients with limited (R-2a), or extensive residual disease (R-2b; log-rank test, P <.0001). CONCLUSION: Cytoreduction followed by HIPEC improves survival in patients with peritoneal carcinomatosis of colorectal origin. However, patients with involvement of six or more regions of the abdominal cavity, or grossly incomplete cytoreduction, had still a grave prognosis. PMID- 14551294 TI - Trends in survival rates after allogeneic hematopoietic stem-cell transplantation for acute and chronic leukemia by ethnicity in the United States and Canada. AB - PURPOSE: Differences in survival among ethnic groups in the United States are reported in numerous diseases and treatment strategies. Whether survival after allogeneic hematopoietic stem-cell transplantation (HSCT) differs by ethnicity is uncertain. PATIENTS AND METHODS: Patients (n = 6443) receiving HLA-identical sibling HSCT for acute or chronic leukemia in the United States or Canada between 1985 and 1999 and reported to the International Bone Marrow Transplant Registry were included. The survival of recipients reported as white, black, Hispanic, or Asian was compared using Cox proportional hazards regression adjusting for other clinical factors. Three 5-year periods were studied to evaluate changes over time. RESULTS: Hispanics compared with whites had lower 1-year (53% v 65%; P <.001) and 3-year adjusted survival rates (38% v 53%; P <.001) between 1995 and 1999, the most recent period studied. We failed to find significant differences in survival rates comparing whites with blacks or with Asians in any of the time periods. Overall survival for the entire cohort improved over time, from 56% to 63% at 1 year and from 43% to 51% at 3 years, with greater improvements noted among blacks (45% to 61% at 1 year and 34% to 48% at 3 years). CONCLUSION: Disparities remain in survival rates between whites and Hispanics despite adjustment for clinical factors. Factors not accounted for in this analysis, such as comorbid disease, socioeconomic status, healthcare access and delivery, and psychosocial and cultural variables, require further prospective study. PMID- 14551296 TI - Patients' perceptions of quality of life after treatment for early prostate cancer. AB - PURPOSE: Treatment for early prostate cancer produces problematic physical side effects, but prior studies have found little influence on patients' perceived health status. We examined psychosocial outcomes of treatment for early prostate cancer. PATIENTS AND METHODS: Patients with previously treated prostate cancer and a reference group of men with a normal prostate-specific antigen (PSA) level and no history of prostate cancer completed questionnaires. Innovative scales assessed behavioral consequences of urinary dysfunction, sexuality, health worry, PSA concern, perceived cancer control, treatment decision making, decision regret, and cancer-related outlook. Urinary, bowel, and sexual dysfunction were assessed with symptom indexes; health status was assessed by the Physical and Mental Summaries of the Short Form (SF-12) Health Survey. RESULTS: Compared with men without prostate cancer, prostate cancer patients reported greater urinary, bowel, and sexual dysfunction, but similar health status. They reported worse problems of urinary control, sexual intimacy and confidence, and masculinity, and greater PSA concern. Perceptions of cancer control and treatment decisions were positive, but varied by treatment: prostatectomy patients indicated the highest and observation patients indicated the lowest cancer control. Bowel and sexual dysfunction were associated with poorer sexual intimacy, masculinity, and perceived cancer control; masculinity and PSA concern were associated with greater confidence in treatment choice; and diminished sexual intimacy and less interest in PSA were associated with greater regret. CONCLUSION: The lack of change in global measures of health status after treatment for early prostate cancer obscures important influences in men's lives; cancer diagnosis and treatment complications may result in complex outcomes. Aggressive treatment may confer confidence in cancer control, yet be countered by diminished intimate relationships and masculinity, which accompany sexual dysfunction. PMID- 14551295 TI - Interferon alfa-2b three times daily and thalidomide in the treatment of metastatic renal cell carcinoma. AB - PURPOSE: The antiangiogenic effect of interferon (IFN) may improve with frequent dosing and by combination with other agents with antiangiogenic activity. To evaluate this potential, we treated patients with metastatic renal cell carcinoma (RCC) with frequently dosed IFN and thalidomide. PATIENTS AND METHODS: Thirty patients were given IFN-alpha-2b 0.9 MU subcutaneously three times daily for 1 month and subsequently 1.2 MU tid unless serious toxicity was encountered. Thalidomide was first given 100 mg/d for 1 week and 300 mg/d thereafter. Sera were collected before and during treatment for serum vascular endothelial growth factor (S-VEGF) analyses performed using enzyme-linked immunosorbent assay. RESULTS: The intention-to-treat response rate was 20% (95% CI, 6% to 34%) and response rate for assessable patients (n = 27) was 22% (95% CI, 6% to 38%). All responses were partial. In addition, 17 patients (63%; 95% CI, 45% to 81%) had stable disease for 3 months or longer. The median time to treatment failure was 7.7 months, and median survival time was 14.9 months. The most common cause of thalidomide discontinuation was neuropathy. S-VEGF levels decreased more in patients who responded to therapy compared with those in patients whose condition had stabilized or who had progressive disease (P =.036). CONCLUSION: The combination of frequently dosed IFN-alpha-2b and low-dose thalidomide is feasible and active in advanced RCC, but the clinical benefit may remain small compared with that of IFN alone. Results from an ongoing phase III trial comparing IFN alpha with or without thalidomide need to be analyzed before this combination can be recommended for use outside clinical studies. PMID- 14551297 TI - Antineoplastic effects of partially HLA-matched irradiated blood mononuclear cells in patients with renal cell carcinoma. AB - PURPOSE: Vaccines, cytokines, and other biologic-based therapies are being developed as antineoplastic agents. Many of these agents are designed to induce an autologous immune response directed against the malignancy. In contrast, hematopoietic stem-cell transplantation is being developed as a form of allogeneic immunotherapy. This study tests the tolerance and antineoplastic activity of sequential infusions of partially HLA-matched allogeneic blood mononuclear cells (obtained from relatives) when administered outside of the context of a hematopoietic stem-cell transplantation. The cells are irradiated to prevent graft-versus-host disease. PATIENTS AND METHODS: Fifteen patients with relapsed or refractory malignancies for which no standard therapy was available were enrolled onto a clinical trial designed to assess the tolerability and antineoplastic effects of irradiated partially HLA-matched blood mononuclear cells obtained from relatives. RESULTS: There was disease regression in three patients with metastatic renal cell carcinoma during treatment. There was disease progression in six patients with metastatic renal cell carcinoma and two patients with metastatic melanoma during treatment. There was no change in disease state in several other patients. CONCLUSION: Irradiated allogeneic blood mononuclear cells administered outside the context of hematopoietic stem-cell transplantation may induce disease responses in patients with relapsed or refractory malignancies. Transfusion of irradiated allogeneic blood mononuclear cells should be developed further as a novel therapeutic antineoplastic approach. PMID- 14551298 TI - Does timing of adjuvant chemotherapy for early breast cancer influence survival? AB - PURPOSE: Theoretically, patients with early breast cancer might benefit from starting adjuvant chemotherapy soon after surgery, and this would have important clinical implications. We have addressed this question from a large, single center database in which the majority of patients received anthracyclines. PATIENTS AND METHODS: A total of 1161 patients from a prospectively maintained database treated with adjuvant chemotherapy for early breast cancer at the Royal Marsden Hospital (London, United Kingdom), including 686 (59%) receiving anthracyclines, were retrospectively analyzed. The disease-free survival (DFS) and overall survival (OS) of the 368 patients starting chemotherapy within 21 days of surgery (group A) were compared with those of the 793 patients commencing chemotherapy >or= 21 days after surgery (group B). Median follow-up time was 39 months (range, 12 to 147 months). RESULTS: No significant difference in 5-year DFS was found between the two groups overall (70% for group A v 72% for group B; P =.4) or in any subgroup. Likewise, there was no difference in 5-year OS (82% for group A v 84% for group B; P =.2) or when the interval to the start of chemotherapy was considered as a continuous variable (P =.4). CONCLUSION: We have been unable to identify any significant survival benefit from starting adjuvant chemotherapy early after surgery, either overall or in any subset of patients. PMID- 14551299 TI - Randomized phase III trial of standard timed doxorubicin plus cisplatin versus circadian timed doxorubicin plus cisplatin in stage III and IV or recurrent endometrial carcinoma: a Gynecologic Oncology Group Study. AB - PURPOSE: To determine if circadian timed (CT) chemotherapy results in improved response, progression-free survival (PFS), overall survival (OS), and lower toxicity, when compared with standard timed (ST) chemotherapy. MATERIALS AND METHODS: Eligibility criteria were stage III, IV, or recurrent endometrial cancer with poor potential for cure by radiation therapy or surgery; measurable disease; and no prior chemotherapy. Therapy was randomized to schedules of ST doxorubicin 60 mg/m2 plus cisplatin 60 mg/m2, or CT doxorubicin 60 mg/m2 at 6:00 am plus cisplatin 60 mg/m2 at 6:00 pm. Cycles were repeated every 3 weeks to a maximum of eight cycles. RESULTS: The ST arm included 169 patients, and the CT arm included 173 patients. The objective response rate (complete responses plus partial responses) was 46% in the ST group compared with 49% in the CT group (P =.26, one tail). Median PFS and OS were 6.5 and 11.2 months, respectively, in the ST group; and 5.9 and 13.2 months, respectively, in the CT group (PFS: P =.31; OS: P =.21, one tail). Median total doses were 209 mg/m2 doxorubicin and 349 mg/m2 cisplatin in the ST group, versus 246 mg/m2 doxorubicin and 354 mg/m2 cisplatin in the CT group. Grade 3 or 4 leukopenia occurred in 73% of patients in the ST arm and in 63% of patients in the CT arm. There were eight treatment-related deaths. CONCLUSION: In this trial, no significant benefit in terms of response rate, PFS or OS, or toxicity profile was observed with CT doxorubicin plus cisplatin in patients with advanced or recurrent endometrial carcinoma. PMID- 14551300 TI - Prognostic significance of p53 mutation and p53 overexpression in advanced epithelial ovarian cancer: a Gynecologic Oncology Group Study. AB - PURPOSE: The prognostic significance of p53 mutations and overexpression in advanced epithelial ovarian cancers was examined in primary tumors from 125 patients participating in a Gynecologic Oncology Group randomized phase III treatment protocol. PATIENTS AND METHODS: Mutational analysis of p53 was performed in RNA or genomic DNA extracted from frozen tumor. An immunohistochemistry assay was used to detect p53 overexpression in fixed tumor. RESULTS: There were 81 patients (74%) with a single mutation, three patients (3%) with two mutations, and 25 patients (23%) lacking a mutation in exons 2 to 11 of p53. Although most mutations occurred within exons 5 to 8, mutations outside this region were observed in 11% of patients. A mutation in exons 2 to 11 of p53 was associated with a short-term improvement in overall survival and progression-free survival. Adjusted Cox modeling demonstrated a 70% reduction in risk of death (P =.014) and a 60% reduction in risk of disease progression (P =.014) for women with such mutations. However, these striking risk reductions increased over time (P <.02) and eventually disappeared with longer follow-up. Overexpression of p53 was observed in 55 patients (100%) with only missense mutation(s), seven patients (32%) with truncation mutations, and eight patients (40%) lacking a mutation in exons 2 to 11. Overexpression of p53 was associated with tumor grade but not with patient outcome. CONCLUSION: Alterations in p53 are a common event in advanced epithelial ovarian cancer. A mutation in p53, but not overexpression of p53, is associated with a short-term survival benefit. Additional studies are required to define the roles that p53 plays in regulating therapeutic responsiveness and patient outcome. PMID- 14551301 TI - Intranodal administration of peptide-pulsed mature dendritic cell vaccines results in superior CD8+ T-cell function in melanoma patients. AB - PURPOSE: We evaluated the feasibility, safety, and immunogenicity of mature, peptide-pulsed dendritic cell (DC) vaccines administered by different routes. PATIENTS AND METHODS: We performed a randomized, phase I, dose-escalation study in 27 patients with metastatic melanoma receiving four autologous peptide-pulsed DC vaccinations. Patients were randomly assigned to an intravenous (IV), intranodal (IN), or intradermal (ID) route of administration (ROA). For each route, primary end points were dose-limiting toxicity, maximum-tolerated dose, and T-cell sensitization. Sensitization was evaluated through tetramer staining, in vitro peptide recognition assays, and delayed-type hypersensitivity (DTH) responses. RESULTS: Twenty-two (81.5%) of 27 patients completed all four vaccinations. Vaccinations were well tolerated; a few patients exhibited grade 1 to 2 toxicities including rash, fever, and injection site reaction. All routes of administration induced comparable increases in tetramer-staining CD8+ T cells (five of seven IV, four of seven IN, and four of six ID patients). However, the IN route induced significantly higher rates for de novo development of CD8+ T cells that respond by cytokine secretion to peptide-pulsed targets (six [85.7%] of seven IN patients v two [33%] of six ID patients v none [0%] of six IV patients; P =.005) and de novo DTH (seven [87.5%] of eight IN patients v two [33.3%] of six ID patients v one [14.3%] of seven IV patients; P =.01) compared with other routes. CONCLUSION: Administration of this peptide-pulsed mature DC vaccine by IN, IV, or ID routes is feasible and safe. IN administration seems to result in superior T-cell sensitization as measured by de novo target-cell recognition and DTH priming, indicating that IN may be the preferred ROA for mature DC vaccines. PMID- 14551302 TI - Association between telomerase activity and outcome in patients with nonmetastatic Ewing family of tumors. AB - PURPOSE: Telomerase is considered a molecular marker for malignancy. The aim of this study was to determine telomerase activity (TA) as a prognostic factor at diagnosis and as a marker for minimal residual disease during therapy and follow up in nonmetastatic Ewing family of tumors (EFT). PATIENTS AND METHODS: Primary tumor specimens and 97 peripheral blood (PBL) samples from 31 EFT patients were analyzed for TA by the Telomeric Repeat Amplification Protocol (TRAP assay). The telomerase catalytic subunit (human telomerase reverse transcriptase [hTERT]) gene expression was evaluated by quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and telomere length was determined by Southern blotting. The presence of the EFT chimeric transcripts was analyzed by RT-PCR. Correlations with progression-free survival were evaluated. RESULTS: At diagnosis, TA in primary tumors did not correlate with outcome. During therapy and follow-up, highly significant correlation was observed between high TA in PBL samples and adverse prognosis (P <.0001). None of the patients harboring low TA progressed, with a long follow-up (median, 60 months) and a progression-free survival (PFS) of 100%. In nine patients, high TA actually could predict relapse, long before overt clinical relapse. The group of patients with high TA and positive RT-PCR had the most adverse outcome; PFS of 20% (P =.0025). TA was found to be a better prognostic factor than RT-PCR and histopathologic response at surgery. CONCLUSION: The results suggest that TA is a significant prognostic variable, superior to the established clinical prognostic parameters during therapy and tumor surveillance. It could be used in combination with RT-PCR for a new risk classification. PMID- 14551303 TI - A phase I study of irinotecan as a 3-week schedule in children with refractory or recurrent solid tumors. AB - PURPOSE: A phase I study was performed to determine the maximum-tolerated dose (MTD) and safety profile of irinotecan (CPT-11) administered as a single intravenous infusion every 3 weeks in children with recurrent or refractory solid tumors. PATIENTS AND METHODS: Eighty-one patients were enrolled, including 48 less heavily, and 33 heavily pretreated patients (cranial irradiation and/or high dose chemotherapy). Children received CPT-11 as a 120-minute infusion at doses ranging from 200 to 720 mg/m2. The dose-limiting toxicities (DLT) on first cycle were determined in both cohorts. RESULTS: One hundred twenty-two cycles and 81 cycles were administered in less heavily, and heavily pretreated patients, respectively. The primary DLT was delayed diarrhea in less heavily pretreated patients, and neutropenia in heavily pretreated patients. MTD was 600 mg/m2 in both cohorts. Grade 3 to 4 neutropenia occurred in 33% and 38% of cycles in less heavily, and heavily pretreated patients, respectively. Grade 3 to 4 nonhematologic toxicities included nausea/vomiting (7% and 4% of cycles in less heavily, and heavily pretreated patients, respectively), asthenia (7% and 4% of cycles, respectively), and delayed diarrhea (6% and 2.5% of cycles, respectively). Four partial responses at 600 mg/m2 (high-grade glioma, neuroblastoma, medulloblastoma, and rhabdomyosarcoma) and 21 minor responses and stable diseases were observed. Pharmacokinetic analysis of CPT-11 and SN-38 was performed in 77 patients. The mean +/- standard deviation (SD) CPT-11 plasma clearance was 20.7 +/- 9.5 L/h/m2 (range, 5 to 54). The mean +/- SD SN-38 metabolic ratio was 1.5% +/- 1.1% (range, 0.15% to 5.55%). CONCLUSION: The recommended phase II dose of CPT-11 in a 3-week schedule is 600 mg/m2 in less heavily, and heavily pretreated children with solid tumors. PMID- 14551304 TI - Early molecular response of marrow disease to biologic therapy is highly prognostic in neuroblastoma. AB - PURPOSE: A promising treatment strategy for stage 4 neuroblastoma patients is the repeated application of anti-GD2 immunotherapy after activating myeloid effectors with granulocyte-macrophage colony-stimulating factor (GM-CSF). To use early marrow response as a prognostic marker is particularly relevant for patients not likely to benefit from this therapy. PATIENTS AND METHODS: Eighty-six stage 4 neuroblastoma patients older than 1 year at diagnosis were classified in four clinical groups on protocol entry: complete remission or very good partial remission (n = 33), primary refractory (n = 33), secondary refractory (n = 10), and progressive disease (n = 10). Bone marrow samples collected before and following treatment were assayed for GD2 synthase mRNA by real-time reverse transcriptase polymerase chain reaction. Response and survival analyses were performed on posttreatment samples before the third cycle at 1.8 months from protocol entry. RESULTS: GD2 synthase mRNA was evident in pretreatment marrow samples of the four clinical groups (42%, 52%, 60%, and 80% of samples, respectively), with median transcript level of 10.0, 16.6, 26.5, and 87.2, respectively. This marker became negative following antibody plus GM-CSF in 77% of complete remission or very good partial remission, 45% of primary refractory, 25% of secondary refractory, and 0% of progressive disease group. Progression free survival was statistically different between responder and nonresponder groups (P <.0001). Among patients with minimal residual disease, molecular responders had a significantly lower risk of disease progression at a median follow-up of 29.8 months (P =.0001). CONCLUSION: GD2 synthase mRNA is a sensitive response marker of neuroblastoma in the bone marrow. It is particularly useful for minimal residual disease evaluation and may potentially be useful as an early predictor of resistance to antibody plus GM-CSF immunotherapy. PMID- 14551305 TI - Dissemination of information on potentially fatal adverse drug reactions for cancer drugs from 2000 to 2002: first results from the research on adverse drug events and reports project. AB - PURPOSE: To describe the clinical findings, occurrence rates, causality evidence, and dissemination media for serious cancer drug-associated adverse drug reactions (ADRs) reported in the postmarketing setting. METHODS: ADRs were termed serious if they resulted in death or severe organ failure. ADR information for oncology drugs from package insert (PI) revisions, so-called Dear Doctor letters, and journal articles was evaluated to identify serious ADRs reported from 2000 to 2002. Timing and content of information disseminated was assessed. RESULTS: Twenty-five serious ADRs associated with 22 oncology drugs were identified after approval. Approximately half of these serious ADRs are associated with drugs approved before 1995. ADRs were described in articles in medical journals (17 ADRs), PI revisions (18 ADRs), and Dear Doctor letters (12 ADRs). PI revisions occurred less than 1 year after peer-reviewed publication for four ADRs. These revisions often differed for similar ADRs that occurred with drugs of the same class. Five of the seven ADRs lacking PI changes occurred with off-label use, for which PI change is not recommended by US Food and Drug Administration (FDA) policy. No cancer drug was withdrawn from the market during the observation period. CONCLUSION: Our findings demonstrate that serious ADRs may be discovered as long as 36 years after a drug receives FDA approval. This suggests a need for continued vigilance and efficient strategies for dissemination of information about ADRs associated with cancer drugs. PMID- 14551307 TI - Unusual cases involving the CNS and nasal sinuses: Case 1. Primary leptomeningeal melanoma. PMID- 14551306 TI - Long-term psychological impact of carrying a BRCA1/2 mutation and prophylactic surgery: a 5-year follow-up study. AB - PURPOSE: To explore long-term psychosocial consequences of carrying a BRCA1/2 mutation and to identify possible risk factors for long-term psychological distress. PATIENTS AND METHODS: Five years after genetic test disclosure, 65 female participants (23 carriers, 42 noncarriers) of our psychological follow-up study completed a questionnaire and 51 participants were interviewed. We assessed general and hereditary cancer-related distress, risk perception, openness to discuss the test result with relatives, body image and sexual functioning. RESULTS: Carriers did not differ from noncarriers on several distress measures and both groups showed a significant increase in anxiety and depression from 1 to 5 years follow-up. Carriers having undergone prophylactic surgery (21 of 23 carriers) had a less favorable body image than noncarriers and 70% reported changes in the sexual relationship. A major psychological benefit of prophylactic surgery was a reduction in the fear of developing cancer. Predictors of long-term distress were hereditary cancer-related distress at blood sampling, having young children, and having lost a relative to breast/ovarian cancer. Long-term distress was also associated with less open communication about the test result within the family, changes in relationships with relatives, doubting about the validity of the test result, and higher risk perception. CONCLUSION: Our findings support the emerging consensus that genetic predisposition testing for BRCA1/2 does not pose major mental health risks, but our findings also show that the impact of prophylactic surgery on aspects such as body image and sexuality should not be underestimated, and that some women are at risk for high distress, and as a result, need more attentive care. PMID- 14551308 TI - Unusual cases involving the CNS and nasal sinuses: Case 2. Sinonasal undifferentiated carcinoma. PMID- 14551310 TI - It's the poison that killed him, you say! PMID- 14551309 TI - From access to evidence: an advocate's journey. PMID- 14551311 TI - c-kit proto-oncogene product is rarely detected in colorectal adenocarcinoma. PMID- 14551314 TI - Chemoradiotherapy in unresectable squamous cell head and neck cancer. PMID- 14551316 TI - Long-term survivors of testicular cancer. PMID- 14551318 TI - Hybridization between Brassica napus and B. rapa on a national scale in the United Kingdom. AB - Measures blocking hybridization would prevent or reduce biotic or environmental change caused by gene flow from genetically modified (GM) crops to wild relatives. The efficacy of any such measure depends on hybrid numbers within the legislative region over the life-span of the GM cultivar. We present a national assessment of hybridization between rapeseed (Brassica napus) and B. rapa from a combination of sources, including population surveys, remote sensing, pollen dispersal profiles, herbarium data, local Floras, and other floristic databases. Across the United Kingdom, we estimate that 32,000 hybrids form annually in waterside B. rapa populations, whereas the less abundant weedy populations contain 17,000 hybrids. These findings set targets for strategies to eliminate hybridization and represent the first step toward quantitative risk assessment on a national scale. PMID- 14551319 TI - A genetic screen in Drosophila for metastatic behavior. AB - A genetic screen was designed in Drosophila to interrogate its genome for mutations sufficient to cause noninvasive tumors of the eye disc to invade neighboring or distant tissues. We found that cooperation between oncogenic RasV12 expression and inactivation of any one of a number of genes affecting cell polarity leads to metastatic behavior, including basement membrane degradation, loss of E-cadherin expression, migration, invasion, and secondary tumor formation. Inactivation of these cell polarity genes cannot drive metastatic behavior alone or in combination with other tumor-initiating alterations. These findings suggest that the oncogenic background of tissues makes a distinct contribution toward metastatic development. PMID- 14551320 TI - Oceanic forcing of Sahel rainfall on interannual to interdecadal time scales. AB - We present evidence, based on an ensemble of integrations with NSIPP1 (version 1 of the atmospheric general circulation model developed at NASA's Goddard Space Flight Center in the framework of the Seasonal-to-Interannual Prediction Project) forced only by the observed record of sea surface temperature from 1930 to 2000, to suggest that variability of rainfall in the Sahel results from the response of the African summer monsoon to oceanic forcing, amplified by land-atmosphere interaction. The recent drying trend in the semiarid Sahel is attributed to warmer-than-average low-latitude waters around Africa, which, by favoring the establishment of deep convection over the ocean, weaken the continental convergence associated with the monsoon and engender widespread drought from Senegal to Ethiopia. PMID- 14551321 TI - Olivo-cerebellar cluster-based universal control system. AB - The olivo-cerebellar network plays a key role in the organization of vertebrate motor control. The oscillatory properties of inferior olive (IO) neurons have been shown to provide timing signals for motor coordination in which spatio temporal coherent oscillatory neuronal clusters control movement dynamics. Based on the neuronal connectivity and electrophysiology of the olivo-cerebellar network we have developed a general-purpose control approach, which we refer to as a universal control system (UCS), capable of dealing with a large number of actuator parameters in real time. In this UCS, the imposed goal and the resultant feedback from the actuators specify system properties. The goal is realized through implementing an architecture that can regulate a large number of parameters simultaneously by providing stimuli-modulated spatio-temporal cluster dynamics. PMID- 14551322 TI - Branched aliphatic alkanes with quaternary substituted carbon atoms in modern and ancient geologic samples. AB - A pseudohomologous series of branched aliphatic alkanes with a quaternary substituted carbon atom (BAQCs, specifically 2,2-dimethylalkanes and 3,3- and 5,5 diethylalkanes) were identified in warm (65 degrees C) deep-sea hydrothermal waters and Late Cretaceous black shales. 5,5-Diethylalkanes were also observed in modern and Holocene marine shelf sediments and in shales spanning the last 800 million years of the geological record. The carbon number distribution of BAQCs indicates a biological origin. These compounds were observed but not identified in previous studies of 2.0 billion- to 2.2 billion-year-old metasediments and were commonly misidentified in other sediment samples, indicating that BAQCs are widespread in the geological record. The source organisms of BAQCs are unknown, but their paleobiogeographic distribution suggests that they have an affinity for sulfides and might be nonphotosynthetic sulfide oxidizers. PMID- 14551323 TI - Site-specific integration of Agrobacterium tumefaciens T-DNA via double-stranded intermediates. AB - Agrobacterium tumefaciens-mediated genetic transformation involves transfer of a single-stranded T-DNA molecule (T strand) into the host cell, followed by its integration into the plant genome. The molecular mechanism of T-DNA integration, the culmination point of the entire transformation process, remains largely obscure. Here, we studied the roles of double-stranded breaks (DSBs) and double stranded T-DNA intermediates in the integration process. We produced transgenic tobacco (Nicotiana tabacum) plants carrying an I-SceI endonuclease recognition site that, upon cleavage with I-SceI, generates DSB. Then, we retransformed these plants with two A. tumefaciens strains: one that allows transient expression of I SceI to induce DSB and the other that carries a T-DNA with the I-SceI site and an integration selection marker. Integration of this latter T-DNA as full-length and I-SceI-digested molecules into the DSB site was analyzed in the resulting plants. Of 620 transgenic plants, 16 plants integrated T-DNA into DSB at their I-SceI sites; because DSB induces DNA repair, these results suggest that the invading T DNA molecules target to the DNA repair sites for integration. Furthermore, of these 16 plants, seven plants incorporated T-DNA digested with I-SceI, which cleaves only double-stranded DNA. Thus, T-strand molecules can be converted into double-stranded intermediates before their integration into the DSB sites within the host cell genome. PMID- 14551324 TI - Multiple hormones act sequentially to mediate a susceptible tomato pathogen defense response. AB - Phytohormones regulate plant responses to a wide range of biotic and abiotic stresses. How a limited number of hormones differentially mediate individual stress responses is not understood. We have used one such response, the compatible interaction of tomato (Lycopersicon esculentum) and Xanthomonas campestris pv vesicatoria (Xcv), to examine the interactions of jasmonic acid (JA), ethylene, and salicylic acid (SA). The role of JA was assessed using an antisense allene oxide cyclase transgenic line and the def1 mutant to suppress Xcv-induced biosynthesis of jasmonates. Xcv growth was limited in these lines as was subsequent disease symptom development. No increase in JA was detected before the onset of terminal necrosis. The lack of a detectable increase in JA may indicate that an oxylipin other than JA regulates basal resistance and symptom proliferation. Alternatively, there may be an increase in sensitivity to JA or related compounds following infection. Hormone measurements showed that the oxylipin signal must precede subsequent increases in ethylene and SA accumulation. Tomato thus actively regulates the Xcv-induced disease response via the sequential action of at least three hormones, promoting expansive cell death of its own tissue. This sequential action of jasmonate, ethylene, and SA in disease symptom development is different from the hormone interactions observed in many other plant-pathogen interactions. PMID- 14551325 TI - Reexamining the role of the accessory plasmid pAtC58 in the virulence of Agrobacterium tumefaciens strain C58. AB - Isogenic strains of Agrobacterium tumefaciens carrying pTiC58, pAtC58, or both were constructed and assayed semiquantitatively and quantitatively for virulence and vir gene expression to study the effect of the large 542-kb accessory plasmid, pAtC58, on virulence. Earlier studies indicate that the att (attachment) genes of A. tumefaciens are crucial in the ability of this soil phytopathogen to infect susceptible host plants. Mutations in many att genes, notably attR and attD, rendered the strain avirulent. These genes are located on pAtC58. Previous work also has shown that derivatives of the wild-type strain C58 cured of pAtC58 are virulent as determined by qualitative virulence assays and, hence, pAtC58 was described as nonessential for virulence. We show here that the absence of pAtC58 in pTiC58-containing strains results in reduced virulence but that disruption of the attR gene does not result in avirulence or a reduction in virulence. Our studies indicate that pAtC58 has a positive effect on vir gene induction as revealed by immunoblot analysis of Vir proteins and expression of a PvirB::lacZ fusion. PMID- 14551326 TI - PRT1 of Arabidopsis is a ubiquitin protein ligase of the plant N-end rule pathway with specificity for aromatic amino-terminal residues. AB - The gene PRT1 of Arabidopsis, encoding a 45-kD protein with two RING finger domains, is essential for the degradation of F-dihydrofolate reductase, a model substrate of the N-end rule pathway of protein degradation. We have determined the function of PRT1 by expression in yeast (Saccharomyces cerevisiae). PRT1 can act as a ubiquitin protein ligase in the heterologous host. The identified substrates of PRT1 have an aromatic residue at their amino-terminus, indicating that PRT1 mediates degradation of N-end rule substrates with aromatic termini but not of those with aliphatic or basic amino-termini. Expression of model substrates in mutant and wild-type plants confirmed this substrate specificity. A ligase activity exclusively devoted to aromatic amino-termini of the N-end rule pathway is apparently unique to plants. The results presented also imply that other known substrates of the plant N-end rule pathway are ubiquitylated by one or more different ubiquitin protein ligases. PMID- 14551328 TI - Microspore separation in the quartet 3 mutants of Arabidopsis is impaired by a defect in a developmentally regulated polygalacturonase required for pollen mother cell wall degradation. AB - Mutations in the QUARTET loci in Arabidopsis result in failure of microspore separation during pollen development due to a defect in degradation of the pollen mother cell wall during late stages of pollen development. Mutations in a new locus required for microspore separation, QRT3, were isolated, and the corresponding gene was cloned by T-DNA tagging. QRT3 encodes a protein that is approximately 30% similar to an endopolygalacturonase from peach (Prunus persica). The QRT3 protein was expressed in yeast (Saccharomyces cerevisiae) and found to exhibit polygalacturonase activity. In situ hybridization experiments showed that QRT3 is specifically and transiently expressed in the tapetum during the phase when microspores separate from their meiotic siblings. Immunohistochemical localization of QRT3 indicated that the protein is secreted from tapetal cells during the early microspore stage. Thus, QRT3 plays a direct role in degrading the pollen mother cell wall during microspore development. PMID- 14551327 TI - Recognition of the Agrobacterium tumefaciens VirE2 translocation signal by the VirB/D4 transport system does not require VirE1. AB - Agrobacterium tumefaciens uses a type IV secretion system to deliver a nucleoprotein complex and effector proteins directly into plant cells. The single stranded DNA-binding protein VirE2, the F-box protein VirF and VirE3 are delivered into host cells via this VirB/D4 encoded translocation system. VirE1 functions as a chaperone of VirE2 by regulating its efficient translation and preventing VirE2-VirE2 aggregation in the bacterial cell. We analyzed whether the VirE1 chaperone is also essential for transport recognition of VirE2 by the VirB/D4 encoded type IV secretion system. In addition, we assayed whether translocation of VirF and VirE3, which also forms part of the virE operon, is affected by the absence of VirE1. We employed the earlier developed CRAFT (Cre recombinase Reporter Assay For Translocation) assay to detect transfer of Cre::Vir fusion proteins from A. tumefaciens into plants, monitored by stable reconstitution of a kanamycin resistance marker, and into yeast, screened by loss of the URA3 gene. We show that the C-terminal 50 amino acids of VirE2 and VirE3 are sufficient to mediate Cre translocation into host cells, confirming earlier indications of a C-terminal transport signal. This transfer was independent of the presence or absence of VirE1. Besides, the translocation efficiency of VirF is not altered in a virE1 mutant. The results unambiguously show that the VirE1 chaperone is not essential for the recognition of the VirE2 transport signal by the transport system and the subsequent translocation across the bacterial envelope into host cells. PMID- 14551329 TI - Analysis of the alternative oxidase promoters from soybean. AB - Alternative oxidase (Aox) is a nuclear-encoded mitochondrial protein. In soybean (Glycine max), the three members of the gene family have been shown to be differentially expressed during normal plant development and in response to stresses. To examine the function of the Aox promoters, genomic fragments were obtained for all three soybean genes: Aox1, Aox2a, and Aox2b. The regions of these fragments immediately upstream of the coding regions were used to drive beta-glucuronidase (GUS) expression during transient transformation of soybean suspension culture cells and stable transformation of Arabidopsis. The expression patterns of the GUS reporter genes in soybean cells were in agreement with the presence or absence of the various endogenous Aox proteins, determined by immunoblotting. Deletion of different portions of the upstream regions identified sequences responsible for both positive and negative regulation of Aox gene expression in soybean cells. Reporter gene analysis in Arabidopsis plants showed differential tissue expression patterns driven by the three upstream regions, similar to those reported for the endogenous proteins in soybean. The expression profiles of all five members of the Arabidopsis Aox gene family were examined also, to compare with GUS expression driven by the soybean upstream fragments. Even though the promoter activity of the upstream fragments from soybean Aox2a and Aox2b displayed the same tissue specificity in Arabidopsis as they do in soybean, the most prominently expressed endogenous genes in all tissues of Arabidopsis were of the Aox1 type. Thus although regulation of Aox expression generally appears to involve the same signals in different species, different orthologs of Aox may respond variously to these signals. A comparison of upstream sequences between soybean Aox genes and similarly expressed Arabidopsis Aox genes identified common motifs. PMID- 14551330 TI - SAGE analysis of transcriptome responses in Arabidopsis roots exposed to 2,4,6 trinitrotoluene. AB - Serial analysis of gene expression was used to profile transcript levels in Arabidopsis roots and assess their responses to 2,4,6-trinitrotoluene (TNT) exposure. SAGE libraries representing control and TNT-exposed seedling root transcripts were constructed, and each was sequenced to a depth of roughly 32,000 tags. More than 19,000 unique tags were identified overall. The second most highly induced tag (27-fold increase) represented a glutathione S-transferase. Cytochrome P450 enzymes, as well as an ABC transporter and a probable nitroreductase, were highly induced by TNT exposure. Analyses also revealed an oxidative stress response upon TNT exposure. Although some increases were anticipated in light of current models for xenobiotic metabolism in plants, evidence for unsuspected conjugation pathways was also noted. Identifying transcriptome-level responses to TNT exposure will better define the metabolic pathways plants use to detoxify this xenobiotic compound, which should help improve phytoremediation strategies directed at TNT and other nitroaromatic compounds. PMID- 14551331 TI - Changes in surface area of intact guard cells are correlated with membrane internalization. AB - Guard cells must maintain the integrity of the plasma membrane as they undergo large, rapid changes in volume. It has been assumed that changes in volume are accompanied by changes in surface area, but mechanisms for regulating plasma membrane surface area have not been identified in intact guard cells, and the extent to which surface area of the guard cells changes with volume has never been determined. The alternative hypothesis-that surface area remains approximately constant because of changes in shape-has not been investigated. To address these questions, we determined surface area for intact guard cells of Vicia faba as they underwent changes in volume in response to changes in the external osmotic potential. We also estimated membrane internalization for these cells. Epidermal peels were subjected to external solutions of varying osmotic potential to shrink and swell the guard cells. A membrane-specific fluorescent dye was used to identify the plasma membrane, and confocal microscopy was used to acquire a series of optical paradermal sections of the guard cell pair at each osmotic potential. Solid digital objects representing the guard cells were created from the membrane outlines identified in these paradermal sections, and surface area, volume, and various linear dimensions were determined for these solid objects. Surface area decreased by as much as 40% when external osmotic potential was increased from 0 to 1.5 MPa, and surface area varied linearly with volume. Membrane internalization was approximated by determining the amount of the fluorescence in the cell's interior. This value was shown to increase approximately linearly with decreases in the cell's surface area. The changes in surface area, volume, and membrane internalization were reversible when the guard cells were returned to a buffer solution with an osmotic potential of approximately zero. The data show that intact guard cells undergo changes in surface area that are too large to be accommodated by plasma membrane stretching and shrinkage and suggest that membrane is reversibly internalized to maintain cell integrity. PMID- 14551332 TI - Interactive effects of jasmonic acid, salicylic acid, and gibberellin on induction of trichomes in Arabidopsis. AB - Leaf trichomes protect plants from attack by insect herbivores and are often induced following damage. Hormonal regulation of this plant induction response has not been previously studied. In a series of experiments, we addressed the effects of artificial damage, jasmonic acid, salicylic acid, and gibberellin on induction of trichomes in Arabidopsis. Artificial damage and jasmonic acid caused significant increases in trichome production of leaves. The jar1-1 mutant exhibited normal trichome induction following treatment with jasmonic acid, suggesting that adenylation of jasmonic acid is not necessary. Salicylic acid had a negative effect on trichome production and consistently reduced the effect of jasmonic acid, suggesting negative cross-talk between the jasmonate and salicylate-dependent defense pathways. Interestingly, the effect of salicylic acid persisted in the nim1-1 mutant, suggesting that the Npr1/Nim1 gene is not downstream of salicylic acid in the negative regulation of trichome production. Last, we found that gibberellin and jasmonic acid had a synergistic effect on the induction of trichomes, suggesting important interactions between these two compounds. PMID- 14551333 TI - Requirement of phosphatidylglycerol for maintenance of photosynthetic machinery. AB - Phosphatidylglycerol (PG) is a ubiquitous component of thylakoid membranes. Experiments with the pgsA mutant of the cyanobacterium Synechocystis sp. PCC6803 defective in biosynthesis of PG have demonstrated an indispensable role of PG in photosynthesis. In the present study, we have investigated the light susceptibility of the pgsA mutant with regard to the maintenance of the photosynthetic machinery. Growth of the mutant cells without PG increased the light susceptibility of the cells and resulted in severe photoinhibition of photosynthesis upon a high-light treatment, whereas the growth in the presence of PG was protected against photoinhibition. Photoinhibition induced by PG deprivation was mainly caused by an impairment of the restoration process. The primary target of the light-induced damage in thylakoid membranes, the D1 protein of photosystem (PS) II was, however, synthesized and degraded with similar rates irrespective of whether the mutant cells were incubated with PG or not. Intriguingly, it was found that instead of the synthesis of the D1 protein, the dimerization of the PSII core monomers was impaired in the PG-deprived mutant cells. Addition of PG to photoinhibited cells restored the dimerization capacity of PSII core monomers. These results suggest that PG plays an important role in the maintenance of the photosynthetic machinery through the dimerization and reactivation of the PSII core complex. PMID- 14551334 TI - Reduced expression of aconitase results in an enhanced rate of photosynthesis and marked shifts in carbon partitioning in illuminated leaves of wild species tomato. AB - Wild species tomato (Lycopersicon pennellii) plants bearing a genetic lesion in the gene encoding aconitase (Aco-1; aconitate hydratase EC 4.2.1.3) were characterized at molecular and biochemical levels. The genetic basis of this lesion was revealed by cloning the wild-type and mutant alleles. The mutation resulted in lowered expression of the Aco-1 transcript and lowered levels of both cytosolic and mitochondrial aconitase protein and activity. After in silico analysis, we concluded that in the absence of a recognizable target sequence, the best explanation for the dual location of this protein is inefficient targeting. Biochemical analysis of leaves of the Aco-1 accession suggested that they exhibited a restricted flux through the Krebs cycle and reduced levels of Krebs cycle intermediates but were characterized by elevated adenylate levels and an enhanced rate of CO2 assimilation. Furthermore, the analysis of both steady-state metabolite levels and metabolic fluxes revealed that this accession also exhibited elevated rates of photosynthetic Suc synthesis and a corresponding increase in fruit yield. Therefore, we conclude that the Krebs cycle normally competes with the Suc synthetic pathway for carbon but is not essential for the supply of energy to fuel the operation of this pathway. PMID- 14551336 TI - Targeted integration of T-DNA into the tobacco genome at double-stranded breaks: new insights on the mechanism of T-DNA integration. AB - Agrobacterium tumefaciens T-DNA normally integrates into random sites in the plant genome. We have investigated targeting of T-DNA by nonhomologous end joining process to a specific double-stranded break created in the plant genome by I-CeuI endonuclease. Sequencing of genomic DNA/T-DNA junctions in targeted events revealed that genomic DNA at the cleavage sites was usually intact or nearly so, whereas donor T-DNA ends were often resected, sometimes extensively, as is found in random T-DNA inserts. Short filler DNAs were also present in several junctions. When an I-CeuI site was placed in the donor T-DNA, it was often cleaved by I-CeuI endonuclease, leading to precisely truncated targeted T DNA inserts. Their structure requires that T-DNA cutting occurred before or during integration, indicating that T-DNA is at least partially double stranded before integration is complete. This method of targeting full-length T-DNA with considerable fidelity to a chosen break point in the plant genome may have experimental and practical applications. Our findings suggest that insertion at break points by nonhomologous end joining is one normal mode of entry for T-DNA into the plant genome. PMID- 14551335 TI - A semidwarf phenotype of barley uzu results from a nucleotide substitution in the gene encoding a putative brassinosteroid receptor. AB - Brassinosteroids (BRs) play important roles throughout plant growth and development. Despite the importance of clarifying the mechanism of BR-related growth regulation in cereal crops, BR-related cereal mutants have been identified only in rice (Oryza sativa). We previously found that semidwarf barley (Hordeum vulgare) accessions carrying the "uzu" gene, called "uzu" barley in Japan, are non-responding for brassinolide (BL). We then performed chemical and molecular analyses to clarify the mechanisms of uzu dwarfism using isogenic line pairs of uzu gene. The response of the uzu line to BL was significantly lower than that of its corresponding normal line. Measurement of BRs showed that the uzu line accumulates BRs, similar to known BR-insensitive mutants. The marker synteny of rice and barley chromosomes suggests that the uzu gene may be homologous to rice D61, a rice homolog of Arabidopsis BR-insensitive 1 (BRI1), encoding a BR receptor protein. A barley homolog of BRI1, HvBRI1, was isolated by using degenerate primers. A comparison of HvBRI1 sequences in uzu and normal barley varieties showed that the uzu phenotype is correlated with a single nucleotide substitution. This substitution results in an amino acid change at a highly conserved residue in the kinase domain of the BR-receptor protein. These results may indicate that uzu dwarfism is caused by the missense mutation in HvBRI1. The uzu gene is being introduced into all hull-less barley cultivars in Japan as an effective dwarf gene for practical use, and this is the first report about an agronomically important mutation related to BRs. PMID- 14551337 TI - Characterization of leachianone G 2"-dimethylallyltransferase, a novel prenyl side-chain elongation enzyme for the formation of the lavandulyl group of sophoraflavanone G in Sophora flavescens Ait. cell suspension cultures. AB - Leachianone G (LG) 2"-dimethylallyltransferase, a novel prenyl side-chain elongation enzyme, was identified in Sophora flavescens Ait. cultured cells. The enzyme transfers a dimethylallyl group to the 2" position of another dimethylallyl group attached at position 8 of LG to form sophoraflavanone G, a branched monoterpenoid-conjugated flavanone characteristic to this plant. This membrane-bound dimethylallyltransferase required Mg2+ (optimum concentration was 10 mm) for the reaction and had an optimum pH of 8.8. It utilized dimethylallyl diphosphate as the sole prenyl donor, and the 2'-hydroxy function in LG was indispensable to the activity. The apparent Km values for dimethylallyl diphosphate and LG were 59 and 2.3 microm, respectively. Subcellular localization of three enzymes that participated in the formation of the lavandulyl group was also investigated by sucrose density gradient centrifugation. Two prenyltransferases, naringenin 8-dimethylallyltransferase and LG 2" dimethylallyltransferase, were localized in the plastids, whereas 8 dimethylallylnaringenin 2'-hydroxylase, which catalyzes the crucial step in the lavandulyl-group formation, was associated with the endoplasmic reticulum. These results suggest the close cooperation between the plastids and the endoplasmic reticulum in the formation of lavandulyl groups. PMID- 14551338 TI - Epigenetic switch from posttranscriptional to transcriptional silencing is correlated with promoter hypermethylation. AB - Changes in the distribution of methylcytosine residues along a transgene locus of tobacco (Nicotiana tabacum) in relation to the type of gene silencing were studied in parental plant leaves, calli, and regenerated plants derived thereof. Parental-silenced HeLo1 (hemizygous for locus 1) plants show posttranscriptional silencing of the residing nptII (neomycin phosphotransferase II) transgene and cytosine methylation restricted to the 3' end and center part of the transcribed region. Here, we report that with an increasing number of cell cycles, DNA methylation changes gradually, and methylation is introduced into the promoter during cell culture and more slowly in vegetatively propagated plants. After 24 months of callus in vitro cultivation, an epigenetic variant, designated locus 1E, was obtained in which cytosine methylation of symmetrical (CG and CNG) sites was almost complete within the 5' end of the nptII-transcribed region and the 35S promoter. Further, methylation of nonsymmetrical sites appeared de novo in the promoter, whereas this type of methylation was significantly reduced in the 3' end of the transcribed region when compared with locus 1. The newly established epigenetic patterns were stably transmitted from calli into regenerated plants and their progeny. The protein and steady-state RNA levels remained low in locus 1E, whereas with nuclear run-on assays, no detectable amounts of primary transcripts were found along the nptII gene, indicating that the methylated promoter became inactivated. The results suggest that a switch between posttranscriptional and transcriptional gene silencing could be a mechanism leading to irrevocable shut down of gene expression within a finite number of generations. PMID- 14551339 TI - Letrozole after tamoxifen for breast cancer--what is the price of success? PMID- 14551340 TI - Beyond tamoxifen--extending endocrine treatment for early-stage breast cancer. PMID- 14551342 TI - Pieces of the preeclampsia puzzle. PMID- 14551341 TI - A randomized trial of letrozole in postmenopausal women after five years of tamoxifen therapy for early-stage breast cancer. AB - BACKGROUND: In hormone-dependent breast cancer, five years of postoperative tamoxifen therapy--but not tamoxifen therapy of longer duration--prolongs disease free and overall survival. The aromatase inhibitor letrozole, by suppressing estrogen production, might improve the outcome after the discontinuation of tamoxifen therapy. METHODS: We conducted a double-blind, placebo-controlled trial to test the effectiveness of five years of letrozole therapy in postmenopausal women with breast cancer who have completed five years of tamoxifen therapy. The primary end point was disease-free survival. RESULTS: A total of 5187 women were enrolled (median follow-up, 2.4 years). At the first interim analysis, there were 207 local or metastatic recurrences of breast cancer or new primary cancers in the contralateral breast--75 in the letrozole group and 132 in the placebo group- with estimated four-year disease-free survival rates of 93 percent and 87 percent, respectively, in the two groups (P< or =0.001 for the comparison of disease-free survival). A total of 42 women in the placebo group and 31 women in the letrozole group died (P=0.25 for the comparison of overall survival). Low grade hot flashes, arthritis, arthralgia, and myalgia were more frequent in the letrozole group, but vaginal bleeding was less frequent. There were new diagnoses of osteoporosis in 5.8 percent of the women in the letrozole group and 4.5 percent of the women in the placebo group (P=0.07); the rates of fracture were similar. After the first interim analysis, the independent data and safety monitoring committee recommended termination of the trial and prompt communication of the results to the participants. CONCLUSIONS: As compared with placebo, letrozole therapy after the completion of standard tamoxifen treatment significantly improves disease-free survival. PMID- 14551343 TI - Autosomal dominant polycystic kidney disease: modifier genes and endothelial dysfunction. PMID- 14551344 TI - Controversial issues in the treatment of hyperkalaemia. PMID- 14551345 TI - Percutaneous coronary interventions in patients with mild to moderate chronic renal failure: to dilate or not to dilate? PMID- 14551346 TI - In the erythropoietin era, can we forget alternative or adjunctive therapies for renal anaemia management? The androgen example. PMID- 14551347 TI - Dosing guidelines for fluconazole in patients with renal failure. PMID- 14551348 TI - Kidney transplants, antibodies and rejection: is C4d a magic marker? AB - The immunohistochemical detection of the complement degradation product C4d in renal allograft biopsies has gained considerable clinical interest in recent years. The accumulation of C4d along peritubular capillaries is generally regarded as a marker for an antibody-mediated allo-response and is associated with poor graft survival. The aim of this review is to discuss histological findings associated with the deposition of C4d. Emphasis is placed on diagnostic and therapeutic implications. Unanswered questions regarding C4d and graft injury are highlighted. PMID- 14551349 TI - Modulation of angiotensin II-mediated signalling by heparan sulphate glycosaminoglycans. AB - BACKGROUND: Heparin and angiotensin-converting enzyme inhibitors can be used as a therapeutic option in diabetic nephropathy (DN). Although the mode of action is poorly understood, both agents may retard the progression of DN. Previously, we demonstrated that angiotensin II (Ang II) has an inhibitory effect on the production of heparan sulphate proteoglycan (HSPG) in mesangial cells (MCs). We have now studied the influence of heparin on the Ang II-induced intracellular Ca(2+) release and activation of nuclear factor kappa B (NF-kappaB). METHODS: Human MCs were isolated from renal cortex and cultivated to measure Ca(2+) influx and NF-kappaB activation. RESULTS: Stimulation of MCs with 100 nM Ang II resulted in a rapid increase in the intracellular Ca(2+) concentration ([Ca(2+)](i)), followed by a decline to baseline level. The addition of heparin resulted in an oscillatory pattern of Ca(2+) influxes upon Ang II stimulation. Whereas the rapid increase in [Ca(2+)](i) was most likely due to release from intracellular stores, oscillations in [Ca(2+)](i) were dependent on the presence of extracellular Ca(2+). Heparin alone did not induce Ca(2+) influx. Both the initial increase and the subsequent oscillations in [Ca(2+)](i) could be blocked by losartan. In MCs with chemically or enzymatically altered membrane-associated heparan sulphate glycosaminoglycan (HS-GAG), Ang II stimulation resulted in [Ca(2+)](i) oscillations. Interestingly, in these cells, the addition of heparin or GAG completely prevented [Ca(2+)](i) oscillations. Heparin inhibited NF-kappaB activation in Ang II-stimulated MCs that expressed either normal or chemically altered GAG. CONCLUSIONS: These findings suggest that alterations in HS-GAG chemistry or metabolism under pathological conditions, such as DN, may have direct functional consequences for the local effect of Ang II. PMID- 14551350 TI - Thrombin stimulates production of fibronectin by human proximal tubular epithelial cells via a transforming growth factor-beta-dependent mechanism. AB - BACKGROUND: Tubulointerstitial fibrosis contributes to the progression of many forms of glomerular disease and to end-stage renal failure. Inflammatory mediators generated during glomerular injury may induce tubulointerstitial lesions by stimulating tubular cells. Thrombin has multiple biological functions in addition to its role in haemostasis and has been detected in the urine of patients with glomerular diseases. The present study investigated whether thrombin can modulate the production of fibronectin (FN) in cultured human proximal tubular epithelial cells (PTEC). METHODS: Cultured PTEC were incubated with or without thrombin to examine the effect of thrombin on FN production in PTEC. FN and transforming growth factor-beta (TGF-beta) levels were measured in culture supernatants by enzyme-linked immunosorbent assay (ELISA). Expression of FN mRNA was analysed by reverse transcriptase-polymerase chain reaction. Effects of thrombin on matrix metabolism were examined by enzyme immunoassay for the detection of secreted matrix metalloproteinase (MMP) and its inhibitors (TIMPs) as well as by zymography. RESULTS: Thrombin stimulated FN secretion in PTEC. Thrombin also stimulated TGF-beta secretion in PTEC in a dose-dependent manner. Expression of FN mRNA by PTEC was augmented by thrombin. The stimulatory effect of thrombin on FN secretion was inhibited by neutralizing antibodies against TGF beta but not by an irrelevant antibody. Thrombin-induced FN secretin was also inhibited by thrombin inhibitors, such as antithrombin III, hirudin and argatroban. Although thrombin stimulated TIMP-1 and -2 secretion by PTEC, the stimulatory effect of thrombin on MMP-2 was not statistically significant. Thrombin did not affect the expression of MMP-2 in zymography studies. CONCLUSIONS: We found that thrombin stimulates FN production in PTEC without causing matrix degradation, an effect that may contribute to the formation of tubulointerstitial fibrosis associated with glomerular disease. The stimulatory effect of thrombin on FN production in PTEC is, at least in part, mediated by TGF beta. PMID- 14551351 TI - Osmoregulation of aldose reductase and sorbitol dehydrogenase in cultivated interstitial cells of rat renal inner medulla. AB - BACKGROUND: Little is known about sorbitol metabolism in renal papillary interstitial cells. For characterization we studied regulation of sorbitol synthesis by aldose reductase (AR) and degradation by sorbitol dehydrogenase (SDH) in papillary interstitial cells. METHODS: Interstitial cells were isolated from rat renal inner medulla to a pure cell fraction. mRNA was isolated from cultivated cells and sorbitol, AR and SDH activity were determined enzymatically in homogenates. RESULTS: Sorbitol concentration in these cells at 300 mosmol/l was 4.4+/-0.3 vs 78+/-3.6 micro mol/g protein at 600 mosmol/l. At steady-state conditions at 300 mosmol/l, AR activity was nearly the same as SDH activity (15.1+/-1.6 vs 16.6+/-2.0 U/g protein). At 600 mosmol/l, AR activity increased to 82.5+/-11.4 U/g protein and SDH activity to 31.5+/-6.0 U/g protein. Studying the time course of enzyme activity after changing osmolarity from 300 to 600 mosmol/l, we found half maximal stimulation after 2-3 (AR) or 3 (SDH) days. The amount of AR-mRNA preceded the rise of enzyme activity, whereas SDH-mRNA was not significantly influenced. Lowering osmolarity from 600 to 300 mosmol/l, enzyme activity decreased to less than half within 2 (AR) or 1 (SDH) day(s). CONCLUSIONS: The results suggest that sorbitol metabolism contributes to handling of osmotic stress in rat renal papillary interstitial cells. PMID- 14551352 TI - Cyclosporin A tubular effects contribute to nephrotoxicity: role for Ca2+ and Mg2+ ions. AB - BACKGROUND: Cyclosporin A (CsA) nephrotoxicity has been attributed primarily to renal haemodynamic alterations caused by afferent arteriolar vasoconstriction. However, CsA nephropathy is also characterized by CsA-induced pre-glomerular disturbances and interstitial injury that may occur independently of haemodynamic changes. Given the high lipophilic activity of CsA, we hypothesized that direct tubular injury is likely to contribute to nephrotoxicity. METHODS: To investigate tubular toxicity of CsA, increasing concentrations of CsA (1, 2.5, 10, 25, 50 and 100 micro g/ml) and its vehicle (cremophor) were added to isolated rat proximal tubules (PT). Cell injury was assessed by lactate dehydrogenase (LDH) release. The role of Ca(2+) ions in tubular toxicity and the effect of calcium channel blockers on CsA toxicity were evaluated by measuring intracellular calcium using the fluorescent dye Fura-2 AM. The role of Mg(2+) ions was assessed using high extracellular Mg(2+) medium (2 mM). RESULTS: Whereas cremophor alone was not toxic to PT, CsA caused PT injury but only at the highest concentration (100 micro g/ml). After 90 min incubation, LDH was 22.5% in control PT and 41.9% in PT treated with 100 micro g/ml CsA (P < 0.001, n = 11). There was a transient increase in intracellular calcium ([Ca(2+)](i)) after CsA administration. A low calcium medium (100 nM) prevented CsA injury to renal tubules. However, verapamil, but not nifedipine, enhanced cell damage. Only nifedipine completely prevented [Ca(2+)](i) increases following CsA. Finally, a high Mg(2+) medium attenuated CsA-induced injury. CONCLUSION: We found that high CsA concentrations caused Ca(2+)- and Mg(2+)-dependent PT injury. Thus, low extracellular Ca(2+) and high Mg(2+) media attenuated CsA-induced tubular injury. Verapamil, but not nifedipine, enhanced CsA tubular toxicity. Therefore, CsA-induced tubular injury may contribute to CsA nephrotoxicity independently of haemodynamic disturbances. PMID- 14551353 TI - Effects of increased intra-abdominal pressure and volume expansion on renal function in the rat. AB - BACKGROUND: The effects of increased intra-abdominal pressure (IAP) and volume expansion on renal function in the rat were studied to gain more knowledge of the oliguria seen during laparoscopic procedures and to reduce the detrimental renal effects of IAP. METHODS: IAP was elevated to 5 or 10 mmHg by insufflation of CO(2) and maintained for 2 h in anaesthetized and mechanically ventilated rats. Rats with normal IAP served as controls. An angiotensin II receptor I antagonist, candesartan, was given as a bolus injection and a 5% volume expansion was achieved by i.v. saline infusion. An angiotensin-converting enzyme (ACE) inhibitor was also given. Renal parameters were the glomerular filtration rate (GFR), urine production, the urinary concentrations of sodium and potassium and the osmolality in the urine. The arterial acid-base balance and blood pressure were also monitored. RESULTS: The GFR deteriorated by 70% during pneumoperitoneum (PP) of 10 mmHg. There was a dramatic drop in sodium excretion (88-97%). With candesartan and elevated IAP, there was a drop in mean arterial pressure (from 90 to 55 mmHg) and the negative renal effects were very pronounced. Renal function was better preserved during elevated IAP in combination with volume expansion. CONCLUSIONS: Capnoperitoneum suppresses renal function, especially in combination with angiotensin II receptor 1 blockade and ACE inhibition. Volume expansion reduces the deleterious effects of PP on renal function during elevated IAP. The results suggest that patients should not be given pharmaceuticals blocking the renin-angiotensin-aldosterone system prior to procedures that may increase IAP. It may be beneficial, however, to reduce angiotensin II tension by volume expansion. PMID- 14551354 TI - Mutational analysis of the xanthine dehydrogenase gene in a Turkish family with autosomal recessive classical xanthinuria. AB - BACKGROUND: Classical xanthinuria is classified into two categories: type I, deficient only in xanthine dehydrogenase (XDH) activity; and type II, deficient in both XDH and aldehyde oxidase. Both types present mainly with renal stones and lead to renal failure in some cases. We studied the molecular basis of xanthinuria in a Turkish family with two affected siblings. METHODS: We examined two brothers aged 1 and 14 years who presented with histories of passing several urinary stones. We measured their serum and urine levels of uric acid and oxypurine, chemically analysed their stones and performed allopurinol loading tests to diagnose the type of xanthinuria. In addition, we studied the coding regions of the XDH gene in family members. RESULTS: In the siblings, serum uric acid was undetectable and serum oxypurine was elevated. Laboratory studies showed that the stones that they passed were composed of xanthine, and both were diagnosed as having classical xanthinuria. The allopurinol loading tests indicated their xanthinuria to be type I. Within the entire coding region of the XDH gene, an A to T base change at nucleotide position 2164 was identified in the siblings, indicating a nonsense substitution from AAG (Lys) to TAG (Tyr) at codon 722. Concerning this novel nonsense mutation, restriction fragment length polymorphism (RFLP) analysis showed that the brothers were both homozygous, while the parents were heterozygous, and this confirmed the autosomal recessive inheritance of the XDH gene mutation. CONCLUSIONS: In a Turkish family, we identified a novel point mutation in the XDH gene responsible for classical type I xanthinuria. That both parents had a history of passing renal stones in spite of being heterozygous for that mutation may indicate that individuals with a heterozygous nonsense XDH mutation are more susceptible to nephrolithiasis than healthy individuals. This raises the point that individuals with a heterozygous XDH mutation may also present with renal stones. PMID- 14551355 TI - Expression of Ras GTPases in normal kidney and in glomerulonephritis. AB - BACKGROUND: Small monomeric Ras GTPases play critical and specific roles in the control of cellular proliferation and apoptosis but the expression of the three Ras isoforms (Ha-Ras, Ki-Ras and N-Ras) in human renal tissue is unknown. This work is an immunohistochemical study of Ras expression in normal renal tissue and in membranous glomerulonephritis (MGN), IgA nephropathy (IgAN) and IgA-negative mesangioproliferative glomerulonephritis (MPGN). METHODS: Formalin-fixed, paraffin-embedded tissue was stained using pan-Ras monoclonal antibody (mAb) and Ras isoform-specific mAb. Detection employed a (DAKO Envision) modified polymer system. RESULTS: The expression of Ras isoforms in normal human kidney was cell specific. For example, N-Ras was detected in tubule epithelial cells but not in glomerular or interstitial cells. Ki-Ras was expressed in mesangial cells, interstitial cells and in proximal convoluted tubule cells (PCT) (particularly localized at brush borders) and in collecting duct cells (CD) (localized to cell membranes) but not in podocytes. Cytoplasmic Ha-Ras was detected in all the above cell types except podocytes. MGN was associated with podocyte expression of all three Ras isoforms and with reduced mesangial cell expression of Ha-Ras and Ki Ras. IgAN was characterized by podocyte expression of Ha-Ras (but not Ki-Ras) and reduced mesangial cell expression of Ki-Ras without alterations in mesangial Ha Ras expression. MPGN was associated with reduced mesangial cell Ha-Ras and Ki-Ras expression without significant podocyte Ras expression. CONCLUSION: These disease specific and isoform-specific alterations in Ras expression may be of significance in pathogenesis and warrant further functional investigation. PMID- 14551356 TI - Does vascular endothelial growth factor (VEGF) play a role in the pathogenesis of minimal change disease? AB - BACKGROUND: Minimal change disease (MCD) is one of the major causes of nephrotic syndrome both in children and adults. The pathogenesis of this condition is not clear and it has been suggested that a plasma permeability factor may play a role. Vascular endothelial growth factor (VEGF), also known as vascular permeability factor, has been thought to be one the factors involved. The aim of this study was thus to investigate the role of VEGF in the pathogenesis of MCD. METHODS: The expression of the gene for VEGF and VEGF receptor-2 (VEGFR-2) was estimated using in situ hybridization in renal biopsy specimens taken from patients with nephrotic syndrome and diagnosed histologically as MCD. The results were compared with those obtained in normal renal tissue. Biopsy specimens from eight patients diagnosed as having MCD were randomly selected for the study. The patients were aged 4-60 years at the time of the biopsy. There were four females and four males. All patients had presented with a nephrotic syndrome, five with recent onset of the disease, two with repeated attacks of the syndrome and one had reduced renal function. RESULTS: The gene expression for VEGF, measured as the proportional glomerular area occupied by autoradiographic grains, was significantly less in the patients with MCD than in controls (1.9 +/- 0.4 vs 4.8 +/- 0.6%, P < 0.0025), whereas the gene expression for VEGFR-2 was no different to controls (1.9 +/- 0.4 vs 2.0 +/- 0.2%). CONCLUSIONS: MCD is associated with a reduction in the expression of the gene for VEGF. As VEGF may play an important role in renal repair and survival, it is postulated that the deficiency, which we have shown, may lead to the dysregulation of the repair process in MCD. PMID- 14551357 TI - Tubular staining of modified C-reactive protein in diabetic chronic kidney disease. AB - BACKGROUND: Serum levels of C-reactive protein (CRP) increase during various atherosclerotic as well as kidney diseases. Whether CRP plays a pathophysiological role or rather serves as a marker is unknown. Here, we investigated the role of CRP in diabetic patients with chronic kidney disease. METHODS: Kidney biopsies from 20 diabetic patients, six with IgA nephropathy and six controls (absence of disease) were stained using a commercially available anti-CRP antibody (clone 8). We characterized this antibody by ELISA and found that it mainly recognized 'modified' CRP (mCRP), the conformational isoform of CRP that occurs after dissociation of the pentameric isomer. RESULTS: A specific CRP signal was observed in the cytoplasma of tubules in 17 out of 20 kidney biopsies from diabetic patients, while all glomeruli, vessels and interstitium stained CRP-negative. This signal was absorbed against the mCRP protein suggesting that the detected tissue-based antigen is more closely related to the mCRP conformer than to the native CRP conformer. Almost all patients (eight out of nine) with severe chronic kidney disease [glomerular filtration rate (GFR) <30 ml/min/1.73 m(2)] strongly stained for the mCRP antigen, whereas only four out of 11 patients with mild and moderate chronic kidney disease (GFR >/=30 ml/min/1.73 m(2)) demonstrated a strong CRP signal. Normal renal tissue and most biopsies with IgA nephropathy were mCRP negative. Severity of histologic changes as assessed by histology score and mCRP staining correlated significantly, but no correlation was evident between tubular mCRP staining and serum levels of CRP or proteinuria. CONCLUSIONS: The present group of diabetic patients showed progressive tubular mCRP staining with declining renal function and increasing severity of histological lesions. Further studies in less proteinuric patients should clarify whether tubular mCRP expression constitutes a progression factor. It also needs to be demonstrated whether mCRP accumulates in tubuli to further stimulate interstitial fibrosis or is mandatory for the resolution of the process. Since mCRP staining was independent of proteinuria we suggest that mCRP is locally produced. PMID- 14551358 TI - Nephropathy associated with heroin abuse in Caucasian patients. AB - BACKGROUND: Renal disease is a complication of heroin addiction. Using renal biopsies in Caucasian patients, we studied the types of nephropathy associated with heroin abuse. METHODS: Nineteen renal biopsies were performed on heroin addicts between January 1993 and December 2001. The indications for renal biopsy included proteinuria with or without renal insufficiency. RESULTS: All 19 patients had serological evidence of hepatitis C virus (HCV) infection, one had hepatitis B virus surface antigen and three were HIV positive. Thirteen patients (68.4%) were found to have membranoproliferative glomerulonephritis (MPGN), 12 with type I and one with type III. Of the remaining patients, two had chronic interstitial nephritis, two had acute proliferative glomerulonephritis, one had amyloidosis and one had granulomatous glomerulonephritis with interstitial nephritis. No apparent decline in the incidence of renal disease was observed. CONCLUSIONS: In this cohort of male Caucasian heroin addicts, HCV-associated MPGN was the most frequent pattern of nephropathy, showing that the nephropathy associated with heroin abuse in Caucasians is not of the focal and segmental glomerulosclerosis type, in contrast to previous reports on African-Americans. This aspect may have important implications for patient management and prognosis. PMID- 14551359 TI - No effect of enalapril on progression in autosomal dominant polycystic kidney disease. AB - BACKGROUND: Angiotensin-converting enzyme (ACE) inhibitors are capable of reducing proteinuria and microalbuminuria with preservation of renal function in diabetic and non-diabetic renal disease. We designed a study investigating the effect of enalapril on the protection of renal function in autosomal dominant polycystic kidney disease (ADPKD). METHODS: We studied 61 normotensive and 28 hypertensive ADPKD patients. The normotensive group participated in a randomized double-blind placebo-controlled study, using enalapril. The hypertensive group was randomized for open label treatment with enalapril or the beta-blocker atenolol. The follow-up was 3 years, and renal function was established repetitively by measuring the clearance of inulin. RESULTS: In the normotensive group, renal function at baseline was 112 +/- 3 ml/min and decreased by -8 +/- 2 ml/min (P < 0.001). The loss of renal function in the patients treated with enalapril or placebo was similar (-7 +/- 3 vs -9 +/- 1 ml/min; P = 0.4). Although blood pressure significantly decreased with enalapril treatment, it had no effect on microalbuminuria. In the hypertensive group, renal function at baseline was 89 +/- 2 ml/min. The mean decline in renal function was -12 +/- 2 ml/min (P < 0.001), and was equal in patients treated with enalapril and those treated with atenolol. The patients treated with atenolol required more additional treatment to control blood pressure, but no difference on microalbuminuria was observed between the two treatments. CONCLUSION: This study was unable to detect a beneficial effect of ACE inhibition on loss of renal function in ADPKD patients. PMID- 14551360 TI - Prevalence of biliary lithiasis in a Sicilian population of chronic renal failure patients. AB - BACKGROUND: The aim of this study was to evaluate the prevalence of biliary lithiasis (BL) and associated risk factors in a population of undialysed patients with chronic renal failure (CRF), and to compare these with findings we had obtained previously in chronic haemodialysis (HD) patients and in subjects from the general population located in the same geographic region. METHODS: A total of 118 CRF patients on conservative treatment were included in the study. In all subjects, we measured several clinical and humoral parameters potentially correlated with BL. Liver and biliary tract ultrasonography was performed with a 3.5 MHz linear probe after at least 12 h of fasting. RESULTS: The prevalence of BL in CRF patients was 22%, which was higher than in the general population (chi(2) = 9.4, P < 0.002) but lower than in HD patients (chi(2) = 25.9, P < 0.0001). Age was similar in the three groups. Body mass index (BMI) was significantly higher in the CRF group than in both HD patients (P < 0.0001) and the general population (P < 0.0001). When the CRF group was divided into subjects with or without BL, the only difference was lower serum calcium levels in the subgroup with BL (P < 0.04). CONCLUSIONS: The prevalence of BL in a Sicilian population of CRF patients was higher than in the general population, but lower than in patients with CRF on chronic HD. Apart from BMI, none of the risk factors traditionally associated with BL in the general population were related to BL in the CRF patients. These data suggest that other factors inherent to kidney pathology contribute to the high prevalence of BL in CRF patients. PMID- 14551361 TI - Bone mineral density, biochemical markers and skeletal fractures in haemodialysis patients. AB - BACKGROUND: End-stage renal disease is often associated with altered bone metabolism. METHODS: In order to investigate the determinant factors of bone mineral density (BMD) and the risk factors of fractures, we studied 70 patients; 26 women (23 post-menopausal) and 44 men, (mean+/-SD) aged 60.5+/-14.3 years, treated by standard haemodialysis (HD) for 6.4+/-6.8 years. Main circulating bone biochemical markers were assessed and BMD was measured with a Lunar DPX densitometer at five sites. BMD results are expressed as a function of age and gender (Z-score). RESULTS: Mean Z-score was markedly decreased at the mid-radius (-2.75+/-1.23) whereas it was normal at the femoral neck (-0.42+/-1.13) and lumbar spine (0.02+/-2.13), and total body (-0.62+/-1.53). Time on HD was negatively correlated to the Z-score at the mid-radius and total body but not at the other sites. Serum intact parathyroid hormone (iPTH), whole PTH or cyclase activating PTH (CAP) and bone-specific alkaline phosphatase concentrations were negatively correlated with Z-scores at all sites. Twenty-one out of 70 patients had sustained a total of 27 fractures since the beginning of dialysis therapy (seven ribs, seven ankles, six vertebrae, three humerus, two wrists and two hips). They had a total body Z-score significantly lower than that of patients without fractures, -1.34+/-1.54 vs -0.37+/-1.46, respectively (P<0.031); however, their Z-scores at the other sites were not different. They were on HD for longer time, 10.4+/-9.5 vs 5.0+/-5.1, respectively (P<0.003), and the relative risk of skeletal fractures was 6.4 times greater after 10 years of HD. The seven patients with rib fractures had a decreased Z-score at most of the sites but not at the mid-radius. Rib fractures but no other fractures were associated with markedly decreased body weight, fat mass and serum leptin levels. CONCLUSIONS: In conclusion, the Z-score at the mid-radius was decreased in HD patients and correlated with high serum PTH but not with fractures. Bone fractures were associated with the time passed on HD and with a low total body Z-score. Rib fractures were frequent and associated with a poor nutritional state. PMID- 14551362 TI - Association between extracellular water, left ventricular mass and hypertension in haemodialysis patients. AB - BACKGROUND: Hypertension and left ventricular hypertrophy (LVH) are present in the majority of patients undergoing haemodialysis (HD). These two pathologies persist after dialysis onset, and pharmacological therapy is often required for adequate control of blood pressure (BP). Although fluid overload is a determinant of hypertension, clinical assessment of this parameter remains difficult and unsatisfactory. Bioimpedance analysis (BIA) spectroscopy and the relative determination of extracellular water (ECW%) may provide a simple and inexpensive tool for investigating fluid overload. We studied 110 patients on thrice-weekly HD to determine whether ECW body content correlates with hypertension and LVH in this patient population. METHODS: Hypertension was determined according to the WHO criteria (office BP >/= 140/90 and/or the use of antihypertensive therapy). Twenty-four hour BP monitoring and echocardiography were performed on midweek inter-HD days. Blood chemistries, dialysis dose (spKt/V) and bioimpedance were analysed on midweek HD days. RESULTS: Hypertension was present in 74.5% of patients. There were no differences for age, spKt/V, haemoglobin, serum creatinine and residual renal function between normotensive and hypertensive patients. Twenty-four hour systolic BP (SBP), 24 h diastolic BP and 24 h pulse pressure were higher in hypertensive patients, in spite of antihypertensive therapy. LVH was present in 61.8% of patients. BIA revealed that ECW% was increased in LVH+ patients (LVH+ = 47.5 +/- 7.9%, LVH- = 42.4 +/- 6.2%, P = 0.01) and in hypertensive patients compared with normotensives (46.5 +/- 7.7% vs 43 +/- 7.2%, P = 0.02). Dry body weights and inter-HD body weight increases did not differ between hypertensive and normotensive patients nor between patients with or without LVH. ECW was correlated with SBP (r = 0.35, P < 0.01) and with left ventricular mass index (LVMi(g/sqm)) (r = 0.49, P < 0.001). A stepwise multiple linear regression model revealed that LVMi(g/sqm) was significantly correlated with ECW%, SBP and male gender (r = 0.65, P < 0.001). CONCLUSIONS: LVH and hypertension are present in a majority of HD patients and they are closely correlated with one another. We found associations between fluid load, measured by BIA and expressed as ECW, and BP and LVM. PMID- 14551363 TI - The high incidence of left atrial appendage thrombosis in patients on maintenance haemodialysis. AB - BACKGROUND: The incidence of intracardiac thrombosis in haemodialysis patients has not been studied. Here we determined the incidence in end-stage renal disease patients on maintenance haemodialysis. METHODS: Transoesophageal echocardiography was performed in 215 patients (125 males, 90 females; mean age 60 +/- 9 years). Any potential candidate with current or past chronic or intermittent atrial fibrillation or with cardiovascular diseases was excluded from the study. RESULTS: Thrombi were found in the left atrial appendages in 71 out of 215 subjects (33%). Based on multiple logistic regression analyses, the probability of finding a thrombus was found to be increased in patients on chronic antiplatelet therapy (odds ratio 4.268) and in those with diabetes mellitus and a low haematocrit (0.22 +/- 0.99 UI/ml) up to 12 h after the start of the dialysis session. CONCLUSIONS: These results suggest that haemodialysis patients receiving the LMWH enoxaparin during dialysis are at risk of bleeding up to 10 h after the injection. PMID- 14551365 TI - Methaemoglobinaemia and haemolysis associated with hydrogen peroxide in a paediatric haemodialysis centre: a warning note. AB - BACKGROUND: Haemodialysis exposes patients to contaminants in the dialysate. The AAMI standards deal only with two disinfectants, chlorine and chloramine. We report an event of methaemoglobinaemia and haemolysis related to an unsuspected disinfection agent. METHODS: Nine children aged 3-17 years undergoing dialysis after reconstruction of our paediatric dialysis unit developed methaemoglobinaemia of 3.1-11%, with a mean reduction in haemoglobin levels of 11.9 +/- 5.9% (P < 0.001). Air bubbles were noted in the bloodlines. The water treatment system (WTS) of the dialysis unit is disinfected when necessary by adding concentrated hydrogen peroxide (HP) to the storage tank and circulating it through the re-circulation loop with draining and subsequent flushings. Total chlorine analysis of the water is performed by DPD-iodide colorimetric method. RESULTS: Dialysis water testing yielded a high chloramine concentration in the storage tank and points- of-use stations (3.08 and 2.06 p.p.m., respectively). However, this finding was not true for the tap water, and it also failed to explain the air bubbles in the dialysis tubing. The concentration of free chlorine was within the recommended range. Further investigation revealed that the WTS was disinfected by the service company during remodelling of the unit, without notification of the hospital staff. Since the DPD-iodide test is not specific, and in effect detects not only total chlorine, but all oxidants capable of oxidizing iodide, we assumed the culprit was residual HP that was inadequately flushed from the water system. CONCLUSIONS: HP used for disinfection of the WTS can pose a serious dialysis risk if not flushed out properly. Total chlorine analysis should be performed before every dialysis session, and positive results should prompt further work-up for other oxidants. The clinical staff must always be involved in decisions regarding any intervention in the dialysis water system. PMID- 14551366 TI - Safety of low-dose spironolactone administration in chronic haemodialysis patients. AB - BACKGROUND: Prevention of cardiovascular diseases is essential in chronic haemodialysis patients. Recently, low-dose spironolactone has been shown to decrease cardiovascular mortality in patients with severe heart failure. However, since haemodialysis patients are prone to hyperkalaemia, a known side effect of spironolactone, this treatment is not used in this population. We performed a study to assess whether low-dose spironolactone (3 x 25 mg/week) could be administered without inducing hyperkalaemia in haemodialysis patients. METHODS: The study design included a 2-week baseline period, followed by a 4-week treatment period in which doses of spironolactone were started at 12.5 mg three times/week for 2 weeks, then increased to 25 mg three times/week, and followed by a 2-week wash-out period. Fourteen patients receiving low-dose spironolactone after each dialysis were compared with 21 haemodialysis patients (control group). RESULTS: Low-dose spironolactone did not change mean serum potassium (4.9 +/- 0.7 vs 4.9 +/- 0.3 mmol/l: control). The mean plasma canrenone level induced by administration of spironolactone 25 mg three times/week in the 14 treated patients was 13 +/- 5.3 ng/ml. Serum aldosterone was not significantly modified by the administration of spironolactone in these patients [before, median 0.35; interquartile range (IQR) 0.11-2.83 nmol/l vs after, median 0.22; IQR 0.12-0.60 nmol/l, NS]. Dietary potassium intake and the use of ion-exchange resin, angiotensin-converting enzyme inhibitors and beta-blockers were similar for the two groups throughout the study. CONCLUSION: This non-randomized and non-blinded study shows that administration of 25 mg spironolactone thrice weekly is not associated with an increased frequency of hyperkalaemia in haemodialysis patients when they are carefully monitored. More studies are required, however, before concluding that spironolactone administration is safe in the chronic haemodialysis population. PMID- 14551367 TI - Is spironolactone safe for dialysis patients? AB - BACKGROUND: Spironolactone is useful in heart failure, but is not given to dialysis patients for fear of hyperkalaemia. This study evaluated the safety of spironolactone administration in haemodialysis patients. METHODS: Fifteen haemodialysis outpatients with mean serum potassium <5.6 mEq/l over the preceding 4 months were treated with spironolactone 25 mg daily for 28 days. Serum potassium was measured before every haemodialysis during the study. Aldosterone and renin were measured at the beginning and end of the study. Patients were monitored for side effects. Data were examined with a paired t-test, with patients serving as their own controls and P < 0.05 considered significant. A sample size of 14 was required to achieve a power of 0.8 and a P = 0.05 to detect a potassium difference of 0.5 +/- 0.6 mEq/l. All patients were analysed as intention-to-treat. RESULTS: The mean potassium level was 4.6 +/- 0.6 mEq/l at baseline and 4.9 +/- 0.9 mEq/l at study completion (P = 0.14). Thirteen patients completed the trial with no potassium levels >6.0 mEq/l. Four patients had potassium levels between 5.5 and 6.0 mEq/l. One patient was withdrawn at day 20 after developing hyperkalaemia (7.6 mEq/l). Another patient was withdrawn at day 25 after missing a dialysis treatment. There were no differences in either baseline or 28 day aldosterone or renin levels (16.8 +/- 28.8 vs 11.7 +/- 6.1 ng/dl and 3.5 +/- 3.9 vs 3.5 +/- 3.5 ng/ml/h, respectively). Infrequent side effects included dry mouth, nosebleed, pruritus, gynecomastia and diarrhoea. No significant leukopenia or anaemia was noted. CONCLUSIONS: Spironolactone may be considered as a treatment option for selected chronic haemodialysis patients with heart disease. PMID- 14551368 TI - Unexpected haemodynamic instability associated with standard bicarbonate haemodialysis. AB - BACKGROUND: The bicarbonate concentration in dialysis fluids for intermittent haemodialysis usually is between 32 and 35 mmol/l. The severity of chronic metabolic acidosis secondary to end-stage renal failure is very variable, however, so that in some patients pre-dialysis acidosis is overcorrected. This study aimed to analyse haemodynamic tolerances to metabolic alkalosis during intermittent haemodialysis. METHODS: In this randomized controlled trial with a single blind, cross-over design, we used dialysis liquids with two different bicarbonate concentrations, 32 (modality A) and 26 (modality B) mmol/l, and in 26 patients, 468 dialysis sessions, compared blood pressure, heart rate, incidence of hypotension and the frequency of corrections required with saline or hypertonic glucose infusions. RESULTS: The results of intradialytic haemodynamic monitoring for modalities A and B, respectively, were: lowest systolic blood pressure 120.8+/-20.8 vs 124.3+/-20.6 mmHg (P < 0.01); mean systolic blood pressure 138.5+/-23.8 vs 144.6+/-24.8 mmHg (P < 0.001); and highest heart rate 73.5+/-12.0 vs 75.8 +/- 12.9 (NS); with modality A, patients had more dialysis sessions with hypotensive episodes (5.55 vs 1.7%, P < 0.05) and required more saline or hypertonic glucose infusions (20.9 vs 13.7% of the dialysis sessions, P < 0.05). CONCLUSIONS: Mild metabolic alkalosis resulting from standard bicarbonate haemodialysis (32 mmol/l) may induce symptomatic hypotension. While normalizing chronic metabolic acidosis is desirable, reducing bicarbonate concentrations should be considered in cases of significant alkalaemia or otherwise untreatable haemodynamic instability. PMID- 14551369 TI - Reduction of circulating microemboli in the subclavian vein of patients undergoing haemodialysis using pre-filled instead of dry dialysers. AB - BACKGROUND: Chronic microembolization that can be demonstrated by pulsed Doppler ultrasound may give rise to pulmonary side-effects during haemodialysis by direct vessel obstruction, increased complement activation or platelet aggregation. The objective of the present investigation was to study whether the use of pre-filled instead of dry dialysers would help to minimize the number of microemboli. METHODS: The study cohort consisted of 23 patients undergoing maintenance haemodialysis. Using a 2 MHz pulsed ultrasound device, the subclavian vein downstream to the dialysis fistula was investigated for 10 min during the dialysis session. The ultrasound examination was performed twice during two successive dialysis sessions, using a pre-filled or a dry dialyser in randomized order. RESULTS: In all patients investigated, numerous microembolic signals (MES) could be observed in the subclavian vein. Treatment with pre-filled dialysers was associated with significantly less MES (82 +/- 94) as compared with dry dialysers (268 +/- 296; P = 0.002). CONCLUSIONS: In comparison to dry dialysers, the use of pre-filled dialysers leads to a significant reduction in microembolization, which may prevent repeated damage to the pulmonary vasculature and, thus, cause less pulmonary damage. PMID- 14551370 TI - Comparison of the new polyethersulfone high-flux membrane DIAPES HF800 with conventional high-flux membranes during on-line haemodiafiltration. AB - BACKGROUND: Current modalities of renal replacement therapy allow only a limited removal of larger, possibly toxic molecules, which accumulate in uraemia. Recently, a haemodiafilter has been made available with the new, high-flux, polyethersulfone-based membrane DIAPES HF800. We performed a study to compare DIAPES HF800 with two conventional high-flux membranes in on-line haemodiafiltration (HDF), with respect to the removal properties for the two marker proteins, beta(2)-microglobulin (beta(2)m, 11.8 kDa) and albumin (66.5 kDa). METHODS: In a prospective, controlled study 10 stable end-stage renal disease patients were randomly allocated to 30 sessions of post-dilutional on line HDF with three types of steam-sterilized membranes: DIAPES HF800, polysulfone and polyamide. Blood flow rate was 250 ml/min and treatment time was 240 min. Pre-treatment beta(2)m and albumin plasma concentrations did not differ between the three groups. The concentration of the two proteins was determined before and after treatment in plasma as well as in the continuously collected haemodiafiltrate. RESULTS: Tolerance of all treatments was very good, without any side-effects for all filters. The mean plasma reduction rate of beta(2)m was 77 +/- 1% for DIAPES HF800 and polysulfone whereas it was 71 +/- 1% for polyamide (P < 0.05). The mean beta(2)m amount removed and found in the haemodiafiltrate per session was 230 +/- 14 mg for DIAPES HF800, 186 +/- 13 mg for polysulfone and 147 +/- 13 mg for polyamide (P < 0.05 between each pair of membranes). The same ranking was obtained for albumin removed and found in haemodiafiltrate per session for the three membranes: 5.7 +/- 0.4, 3.5 +/- 0.4 and 1.0 +/- 0.4 g, respectively. Although DIAPES HF800 showed the highest value for albumin in haemodiafiltrate the mean post-treatment plasma albumin was higher after the treatment with DIAPES HF800 compared with the other membranes (P < 0.05). CONCLUSIONS: On-line HDF has shown to achieve plasma reduction rates for beta(2)m of up to 77% for the DIAPES HF800 membrane and for polysulfone. The amounts of beta(2)m and albumin in haemodiafiltrate were much higher for DIAPES HF800 than for the other two membranes indicating a greater permeability for molecules up to a molecular weight of 66.5 kDa. This could, at least theoretically, offer the advantage also to remove uraemic toxins in the molecular weight range of albumin or of albumin-bound toxins. The future must show whether this will counterbalance the loss of albumin. PMID- 14551371 TI - Onset of arterial 'steal' following proximal angioaccess: immediate and delayed types. AB - BACKGROUND: Critical hand ischaemia following angioaccess is a potentially devastating complication and timely surgical repair is necessary to prevent permanent sequelae. However, the duration of the post-operative surveillance needed to exclude its occurrence has not been determined. METHODS: A retrospective review conducted over a 10-year period revealed 28 patients with critical hand ischaemia following access and surgical repair. The initial access that resulted in the limb-threatening 'steal' included 10 autologous brachiocephalic arteriovenous (AV) fistulae and 18 AV bridge grafts. The two groups of patients, those with autologous AV fistulae and those with AV bridge grafts, were compared regarding the time elapsed from the initial access to the correction procedure, the systolic pressure index between the two forearms and the existence of tissue loss. RESULTS: There was a highly significant difference in the time elapsed from the creation of the initial access to the revision procedure among the two groups, the median time being 2 days in the AV graft group and 165 days in the autologous group (P < 0.00001). The method of treatment was the distal revascularization-interval ligation (DRIL) procedure in the majority of patients (23 of 28), with immediate relief of ischaemic symptoms in all and a mid-term 1 year patency of 69%. CONCLUSIONS: Severe steal develops immediately following AV bridge grafting and patients should be closely monitored during the first 24 h; surveillance is not indicated beyond 1 month. In contrast, steal following formation of proximal autogenous fistulae may be either of immediate or of 'late' onset, months or years after the creation of the fistulae, and lifelong monthly surveillance is recommended. Close monitoring is also recommended after any subsequent surgical or interventional correcting procedure for all access types. DRIL is the procedure of choice in limb-threatening severe steal. PMID- 14551372 TI - Intraperitoneal hydrostatic pressure and flow characteristics of peritoneal catheters in automated peritoneal dialysis. AB - BACKGROUND: In automated peritoneal dialysis (APD) one of the most important factors that influence the efficiency of the treatment is the total volume of dialysate infused per session and the dwell time. This study is aimed at examining the relationships between i.p. pressure (IPP), dialysate flow characteristics, and different dialysate fill volumes in order to optimize APD. METHODS: We studied 20 patients who received APD, with the standard fill volume (2 l, A), or individualized fill volumes based on the patient's body surface area (2.5 l/BSA/1.73 m, B) or on body weight (40 ml/kg body weight, C). The patient's tolerance to a given fill volume was evaluated by measuring IPP, and catheter flow characteristics were evaluated by an automated machine. RESULTS: IPP increased with the increase of the infused volume of dialysate (P < 0.05) and tended towards a positive relationship with the patient's body mass index (BMI: A vs IPP: R = 0.39, P = 0.0019; B vs IPP: R = 0.66, P = 0.0012; C vs IPP R = 0.55, P = 0.009). We also found a relationship between fill volume, BMI and IPP: IPP = 1.0839 + 0.53 (beta) x BMI + 0.211 (beta) x fill volume (R = 0.65; r(2) = 0.40 P < 0.01). The mean IPP with different dialysate fill volumes tended to be related to the volume of dialysate drained at the transition point (R = 0.37; P < 0.05). The pre-transition flow rate/mean IPP ratio tended towards a positive relationship with the volume of dialysate drained at the transition point (R = 0.35, P < 0.05), the transition time (R = 0.34; P < 0.05) and a negative one with the transition volume (R = -0.35, P = 0.05). CONCLUSION: It is possible to customize APD, where the tidal percentage coincides with the transition point for a given catheter and a specific initial dialysate fill volume, the tolerance of which can be measured by assessing IPP. PMID- 14551373 TI - Renal transplantation in patients with primary immunoglobulin A nephropathy. AB - BACKGROUND: Opinions on the clinical course and outcome of renal transplantation in patients with primary immunoglobulin A nephropathy (IgAN) have been controversial. METHODS: We conducted a retrospective single-centre study on 542 kidney transplant recipients over the period 1984-2001. Long-term outcome and factors affecting recurrence in recipients with primary IgAN were analysed. RESULTS: Seventy-five patients (13.8%) had biopsy-proven IgAN as the cause of renal failure, and their mean duration of follow-up after transplantation was 100 +/- 5.8 months. Fourteen (18.7%) of the 75 patients had biopsy-proven recurrent IgAN, diagnosed at 67.7 +/- 11 months after transplantation. The risk of recurrence was not associated with HLA DR4 or B35. Graft failure occurred in five (35.7%) of the 14 patients: three due to IgAN and two due to chronic rejection. Three (4.9%) of the 61 patients without recurrent IgAN had graft failure, all due to chronic rejection. Graft survival was similar between living-related and cadaveric/living-unrelated patients (12-year graft survival, 88 and 72%, respectively, P = 0.616). Renal allograft survival within the first 12 years was better in patients with primary IgAN compared with those with other primary diseases (80 vs 51%, P = 0.001). Thereafter, IgAN patients showed an inferior graft survival (74 vs 97% in non-IgAN patients, P = 0.001). CONCLUSIONS: Our data suggested that around one-fifth of patients with primary IgAN developed recurrence by 5 years after transplantation. Recurrent IgA nephropathy in allografts runs an indolent course with favourable outcome in the first 12 years. However, the contribution of recurrent disease to graft loss becomes more significant on long-term follow up. PMID- 14551374 TI - Removal of persisting isohaemagglutinins with Ig-Therasorb immunoadsorption after major ABO-incompatible non-myeloablative allogeneic haematopoietic stem cell transplantation. AB - BACKGROUND: Major ABO-incompatibility can be associated with haemolysis, prolonged red cell aplasia (PRCA) and higher peri-transplant mortality resulting from organ toxicity after conventional and non-myeloablative allogeneic haematopoietic stem cell transplantation. Different therapeutic strategies have been developed to allow erythroid reconstitution in these patients. METHODS: We present three patients, who developed PRCA after non-myeloablative allogeneic haematological stem cell transplantation for haematological malignancies. The patients were treated with Ig-Therasorb immunoadsorption (five treatments per week) to remove persisting incompatible isohaemagglutinins. RESULTS: Two patients became transfusion independent after 12 and 14 treatments. In one patient, however, no reduction of the isohaemagglutinin titres could be observed after 25 treatments, probably due to persistence of his underlying disease. CONCLUSIONS: Although Ig-Therasorb immunoadsorption was effective in only two patients, it seems to be a promising therapeutic option for patients with PRCA after allogeneic non-myeloablative haematological stem cell transplantation. PMID- 14551375 TI - Pharmacokinetic interaction between corticosteroids and tacrolimus after renal transplantation. AB - BACKGROUND: Tacrolimus is an immunosuppressive drug that is a substrate of cytochrome P450 3A (CYP3A) enzymes and P-glycoprotein (P-gp). After transplantation, many pharmacological interactions have been described. Corticosteroids induce both CYP3A and P-gp activity. This study was designed to investigate the presence of a clinically significant interaction between steroids and tacrolimus after renal transplantation. METHODS: We studied 83 renal transplant recipients receiving tacrolimus after transplantation. Patients were divided into three groups, according to steroid dose (low: 0-0.15 mg/kg/day; intermediate: 0.16-0.25 mg/kg/day; and high: >0.25 mg/kg/day). All other medications, including those known to interact with CYP3A and/or P-gp, were recorded. Steroid dosage, tacrolimus dosage, tacrolimus trough concentration (C0) and tacrolimus concentration/dose ratio [C0 divided by the 24 h dosage (mg/kg)] were assessed for each dosage group after 1 and 3 months of tacrolimus treatment. RESULTS: The three groups were not different as regards the use of non immunosuppressive treatments or clinical events. At 1 and 3 months, the tacrolimus doses and concentration/dose ratios differed significantly in the three steroid dosage groups. With the higher doses, higher tacrolimus doses were needed to achieve the blood tacrolimus targeted trough level. CONCLUSIONS: We demonstrated that pharmacokinetic interaction occurs between steroids and tacrolimus in renal transplant patients. The higher the steroid dosage, the higher the dosage of tacrolimus needed to achieve target trough levels in these patients. The most likely interaction mechanism is specific enzymatic induction of CYP3A and/or P-gp. Interaction is present, even when the steroid dosage is low. The clinical events liable to occur during steroid sparing or tapering must be taken into account because it may be associated with episodes of tacrolimus related nephrotoxicity. PMID- 14551376 TI - Impact of vitamin E on plasma asymmetric dimethylarginine (ADMA) in chronic kidney disease (CKD): a pilot study. AB - BACKGROUND: Asymmetric dimethylarginine (ADMA), an endogenous inhibitor of endothelial nitric oxide synthase and a proposed cardiovascular risk factor, is elevated in chronic kidney disease (CKD). Pharmacological strategies that lower plasma concentration of ADMA may be expected to increase nitric oxide (NO.) bioavailability and potentially limit atherosclerosis. We hypothesized that the antioxidant alpha-tocopherol (vitamin E) reduces ADMA levels in CKD. METHODS: An open-label pilot interventional study using 800 IU of vitamin E was undertaken in eight stable out-patients with non-diabetic CKD (creatinine clearance <30 ml/min/1.73 m(2)) and six healthy controls, with the objective of measuring plasma ADMA levels at baseline and after 8 weeks of treatment. Plasma ADMA, symmetric dimethylarginine (SDMA) and alpha-tocopherol concentrations were determined at study entry and exit using high-performance liquid chromatography, while plasma total F2-isoprostanes, an index of oxidative stress, were measured using a commercially available enzyme-linked immunosorbent assay kit. RESULTS: ADMA and SDMA concentrations were significantly higher in the plasma of patients compared with that of controls (P or =20 degrees C), significant positive associations were found between levels of PM10, NO2, SO2, CO, and O3 and both primary intracerebral hemorrhage and ischemic stroke admissions. On cool days (<20 degrees C), only CO levels and ischemic stroke admissions were significantly associated. For the 2-pollutant models, PM10 and NO2 remained consistently and significantly associated with admissions for both types of stroke on warm days. We observed estimated relative risks of 1.54 (95% confidence interval [95%], 1.31 to 1.81) and 1.56 (95% CI, 1.32 to 1.84) for primary intracerebral hemorrhage for each interquartile range increase in PM10 and NO2. The values for ischemic stroke were 1.46 (95% CI, 1.32 to 1.61) and 1.55 (95% CI, 1.40 to 1.71), respectively. The effects of CO, SO2, and O3 were mostly nonsignificant when either NO2 or PM10 was controlled for. CONCLUSIONS: This study provides an association between exposure to air pollution and hospital admissions for stroke. PMID- 14551400 TI - Costs of acute care of first-ever ischemic stroke in Taiwan. AB - BACKGROUND AND PURPOSE: We sought to investigate the direct costs of acute hospitalization for patients with first-ever ischemic stroke in Taiwan. METHODS: Data were prospectively collected from 360 first-ever ischemic stroke patients. Hospital charges were used for analysis. Multiple linear regression analysis was used to identify the main factors influencing costs. RESULTS: Mean age was 64.9 years (median, 67.0 years), and 58% were male. Mean National Institutes of Health Stroke Scale (NIHSS) score at admission was 9.4 (median, 6.0). Mean initial score of modified Barthel Index was 10.7 (median, 12.0). Median length of stay was 7 days (range, 1 to 122 days). In-hospital mortality was 8%. Overall, median cost per patient was 26 326 New Taiwan dollars (NTD) (original currency) or 841 dollars; median cost per day was 3777 NTD or 121 dollars. Median costs for patients with initial NIHSS score 0 to 6, 7 to 15, and 16 to 38 were 20 365 NTD (650 dollars), 31 954 NTD (1020 dollars), and 62 653 NTD (2000 dollars), respectively. Daily component (physician and ward charges) accounted for approximately 38% of total costs. Initial NIHSS score, small-vessel occlusion, admission to intensive care unit, sex, and smoking had significant impacts on costs. CONCLUSIONS: Apart from providing cost estimates, we note that stroke severity strongly affects costs. PMID- 14551401 TI - Interactions between p38 mitogen-activated protein kinase and caspase-3 in cerebral endothelial cell death after hypoxia-reoxygenation. AB - BACKGROUND AND PURPOSE: The emerging concept of the neurovascular unit in stroke reemphasizes the need to focus on endothelial responses in brain. In this study we examined the role of mitogen-activated protein (MAP) kinase signaling in the regulation of hypoxic cell death in cerebral endothelial cells. METHODS: Human cerebral microvascular endothelial cells were exposed to 4 to 12 hours of hypoxia followed by 12 to 24 hours of reoxygenation. Cytotoxicity was measured by quantifying lactate dehydrogenase release. DNA laddering and caspase-3 activity were assessed to document a role for caspase-dependent cell death. zVAD-fmk and zDEVD-fmk were used to inhibit caspases. Activation of extracellular signal regulated kinase (ERK), p38, and c-Jun N-terminal kinase (JNK) was assessed with Western blotting and kinase activity assays. U0126, SB203580, and SP600125 were used to interrupt the ERK, p38, and JNK pathways, respectively. RESULTS: Endothelial cell death occurred primarily during reoxygenation. DNA laddering and caspase activation were observed, and cytotoxicity was ameliorated by caspase inhibitors (20 micromol/L of zVAD-fmk or zDEVD-fmk). Among the 3 major MAP kinases, only p38 was transiently activated during reoxygenation, and inhibition with 10 micromol/L of SB203580 significantly reduced cytotoxicity. No effects were observed with other MAP kinase inhibitors. Cytoprotection with SB203580 was not accompanied by caspase downregulation. In contrast, cytoprotection with zVAD fmk was associated with a decrease in p38 activation. Furthermore, cleavage of MEKK1 (an upstream kinase of p38) was significantly reduced by zVAD-fmk. CONCLUSIONS: Cerebral endothelial cell death after hypoxia-reoxygenation is mediated by interactions between caspases and p38 MAP kinase. Surprisingly, p38 pathways lie downstream of caspase mechanisms in this model system. PMID- 14551402 TI - Racial disparities in receipt of secondary stroke prevention agents among US nursing home residents. AB - BACKGROUND AND PURPOSE: Although medications can significantly reduce the risk of recurrent stroke, little is known about the extent to which such therapies are given to nursing home residents. We sought to evaluate the extent to which people of color were less likely to receive pharmacological agents in the treatment of recurrent stroke while living in US nursing homes. METHODS: We identified 19 051 residents with a recent hospitalization and primary discharge diagnosis of 434 or 436 in 5 states from 1992 to 1996; of these, 7053 had concomitant conditions indicating anticoagulant therapy. We considered aspirin, dipyridamole, ticlopidine, or warfarin alone or in combination as secondary drug prevention. Generalized linear models provided estimates of the absolute difference in prevalence estimates of the receipt of agents used for the prevention of recurrent stroke between each race-ethnicity group adjusted for potential confounders. RESULTS: Variability in use of any treatment was observed by race ethnicity ranging from 58% of American Indians receiving therapy to only 39% of Asian/Pacific Islanders. Among residents with an indication for anticoagulant therapy, the absolute estimated crude differences indicated that residents of color were less likely than non-Hispanic whites to receive warfarin. After controlling for confounding, Asian/Pacific Islanders, blacks, and Hispanics eligible for anticoagulant therapy received warfarin less often than non-Hispanic white residents. CONCLUSIONS: Overall, only half of our elderly population received any pharmacological agent for secondary prevention of stroke. Interventions designed to improve the pharmacological management of recurrent stroke regardless of race are needed in the nursing home setting. PMID- 14551403 TI - Editorial comment--North, south: changing directions in cardiovascular epidemiology. PMID- 14551404 TI - Johannesburg + 1: S and T challenges. PMID- 14551405 TI - Infectious diseases. New biodefense splurge creates hotbeds, shatters dreams. PMID- 14551406 TI - Research oversight. Panel seeks to balance science and security. PMID- 14551407 TI - Russia's nuclear cities. Swords-to-plowshares program suffers meltdown. PMID- 14551408 TI - Cosmology. Polyhedral model gives the universe an unexpected twist. PMID- 14551409 TI - Climate change. Warming Indian Ocean wringing moisture from the Sahel. PMID- 14551410 TI - Scientific publishing. U.S. license needed to edit Iranian papers. PMID- 14551411 TI - Biotechnology. The ultimate gene gizmo: humanity on a chip. PMID- 14551413 TI - Deep-sea drilling. Same crew to run new program. PMID- 14551412 TI - Infectious diseases. SARS researchers report new animal models. PMID- 14551414 TI - Cell biology. How cells step out. PMID- 14551415 TI - Yoshihide Hayashizaki profile.He's writing the book on the mouse genome. PMID- 14551416 TI - Molecular medicine. NIH dives into drug discovery. PMID- 14551417 TI - Ecology. Freshwater eels are slip-sliding away. PMID- 14551419 TI - Assessing the future hydrogen economy. PMID- 14551418 TI - Infectious diseases. An obscure weapon of the cold war edges into the limelight. PMID- 14551420 TI - Perceptions of science. The anatomical mission to Burma. AB - Until the 1830s, most Americans were unfamiliar with the images of anatomy. Then a small vanguard of reformers and missionaries began to preach, at home and around the world, that an identification with the images and concepts of anatomy was a crucial part of the civilizing process. In his essay, Sappol charts the changes in the perception of self that resulted from this anatomical evangelism. Today, as anatomical images abound in the arts and the media, we still believe that anatomical images show us our inner reality. PMID- 14551422 TI - Chemistry. Enhanced: a view of unusual peroxides. AB - Chemical processes involving oxygen can lead to unstable peroxide molecules such as three-membered ring dioxiranes. In his Perspective, Greer describes the stability of a variety of heteroatom-containing dioxiranes and the tools used to characterize unstable peroxides species. He highlights the report by Ho et al., who have used low-temperature nuclear magnetic resonance to obtain direct evidence for dioxirane intermediates. The method may also be applicable to biological systems in which peroxides have been implicated. PMID- 14551421 TI - Circadian rhythms. Liver regeneration clocks on. AB - In some simple protozoans and unicellular algae, the cell cycle clock is strongly influenced by the circadian clock, such that mitotic cell division takes place only at certain times of the day. Now, as Schibler reports in his Perspective, new work on regenerating liver cells in mice (Matsuo et al.) reveals that the cell cycle clock of mammalian cells is also under the yoke of the master circadian oscillator. PMID- 14551423 TI - Atmospheric science. Ozone and climate change. AB - Over the past 40 years, Southern Hemisphere circumpolar westerly winds have strengthened. In his Perspective, Karoly highlights the modeling study by Gillett and Thompson, who show that these observed Southern Hemisphere climate changes in spring and summer can be explained as a response to stratospheric ozone depletion over Antarctica. The observed strengthening of the circumpolar westerlies in winter is less likely to be the response to springtime Antarctic ozone depletion, but may be due in part to increasing atmospheric greenhouse gases. Understanding the different causes and practical impacts of these trends in Southern Hemisphere circulation is an important next step for climate researchers. PMID- 14551424 TI - Neuroscience. Feeling the pain of social loss. AB - Poets have long waxed lyrical about the pain of a broken heart. Now, as Panksepp explains in his Perspective, this metaphor may reflect real events in the mammalian brain. A new brain neuroimaging study (Eisenberger et al.) reveals that the brain areas that are activated during the distress caused by social exclusion are also those activated during physical pain. Thus, we now have an explanation for the feeling of physical pain that accompanies emotional loss-whether that be the loss of a loved one, rejection by one's social group, or the distress of separation experienced by young animals. PMID- 14551425 TI - Chemistry. The motions of an enzyme soloist. AB - Dynamics of proteins are crucial to their function. In his Perspective, Orrit stresses the advantages of studying these dynamics with single-molecule methods- which require no synchronization--rather than with conventional ensemble measurements. He highlights the report by Yang et al., who follow the fluorescence of a single enzyme molecule. Electron transfer from the fluorophore to a quencher induces fluctuations of the fluorescence lifetime along with the fluorophore-quencher distance. The wide range of characteristic times of those fluctuations reveals the complexity of the protein's potential energy landscape. As a new molecular ruler, electron transfer complements other single-molecule methods such as energy transfer (FRET) for distances shorter than a few nanometers. PMID- 14551426 TI - Genomics. Microarrays--guilt by association. AB - DNA microarray analysis has provided a wealth of data on global patterns of gene expression but has yet to deliver on its early promise of identifying networks of interacting gene products. In his Perspective, Quackenbush discusses new work (Stuart et al.) that uses evolutionary conservation of gene expression patterns in yeast, worm, fruit fly, and human in an attempt to identify functionally related groups of genes. PMID- 14551427 TI - Developmental biology. Rocks that roll zebrafish. AB - The vestibular organs of the inner ear of higher vertebrates control balance, and their counterparts in fish control both balance and hearing. Essential to the operation of these sensory organs are the biomineralized structures--otoconia in higher vertebrates or otoliths in fish--that deflect the sensory hair bundles situated beneath them. In her Perspective, Fekete explores the fascinating world of otolith biomineralization in zebrafish; revealing the importance of a protein called Starmaker for coordinating the shape and type of crystal in fish otoliths ( Sollner et al.). PMID- 14551428 TI - The elusive Bose metal. AB - The conventional theory of metals is in crisis. In the past 15 years, there has been an unexpected sprouting of metallic states in low-dimensional systems, directly contradicting conventional wisdom. For example, bosons are thought to exist in one of two ground states: condensed in a superconductor or localized in an insulator. However, several experiments on thin metal-alloy films have observed that a metallic phase disrupts the direct transition between the superconductor and the insulator. We analyze the experiments on the insulator superconductor transition and argue that the intervening metallic phase is bosonic. All relevant theoretical proposals for the Bose metal are discussed, particularly the recent idea that the metallic phase is glassy. The implications for the putative vortex-glass state in the copper oxide superconductors are examined. PMID- 14551429 TI - Structure of West Nile virus. PMID- 14551431 TI - Protein conformational dynamics probed by single-molecule electron transfer. AB - Electron transfer is used as a probe for angstrom-scale structural changes in single protein molecules. In a flavin reductase, the fluorescence of flavin is quenched by a nearby tyrosine residue by means of photo-induced electron transfer. By probing the fluorescence lifetime of the single flavin on a photon by-photon basis, we were able to observe the variation of flavin-tyrosine distance over time. We could then determine the potential of mean force between the flavin and the tyrosine, and a correlation analysis revealed conformational fluctuation at multiple time scales spanning from hundreds of microseconds to seconds. This phenomenon suggests the existence of multiple interconverting conformers related to the fluctuating catalytic reactivity. PMID- 14551430 TI - Phosphadioxirane: a peroxide from an ortho-substituted arylphosphine and singlet dioxygen. AB - We prepared the primary adduct for the reaction of singlet dioxygen (1O2) with an arylphosphine by using the sterically hindered arylphosphine tris(o methoxyphenyl)phosphine. The resulting phosphadioxirane has a dioxygen molecule triangularly bound to the phosphorus atom. Olefin trapping experiments show that the phosphadioxirane can undergo nonradical oxygen atom-transfer reactions. Under protic conditions, two different intermediates are formed during the reaction of singlet dioxygen with tris(o-methoxyphenyl)phosphine, namely, the corresponding hydroperoxy arylphosphine and a hydroxy phosphorane. Experiments with other arylphosphines possessing different electronic and steric properties demonstrate that the relative stability of the tris(o-methoxyphenyl)phosphadioxirane is due to both steric and electronic effects. PMID- 14551432 TI - Periodic mesoporous organosilicas containing interconnected [Si(CH2)]3 rings. AB - A periodic mesoporous organosilica composed of interconnected three-ring [Si(CH2)]3 units built of three SiO2(CH2)2 tetrahedral subunits is reported. It represents the archetype of a previously unknown class of nanocomposite materials in which two bridging organic groups are bound to each silicon atom. It can be obtained with powder and oriented film morphologies. The nanocomposite is self assembled from the cyclic three-ring silsesquioxane [(EtO)2Si(CH2)]3 precursor and a surfactant mesophase to give a well-ordered mesoporous framework. Low dielectric constants and good mechanical stability of the films were measured, making this material interesting for microelectronic applications. Methylene group reactivity of the three-ring precursor provides entry to a family of nanocomposites, exemplified by the synthesis and self-assembly of [(EtO)2Si(CHR)][(EtO)2Si(CH2)]2 (where R indicates iodine, bromine, or an ethyl group). PMID- 14551433 TI - Simulation of recent southern hemisphere climate change. AB - Recent observations indicate that climate change over the high latitudes of the Southern Hemisphere is dominated by a strengthening of the circumpolar westerly flow that extends from the surface to the stratosphere. Here we demonstrate that the seasonality, structure, and amplitude of the observed climate trends are simulated in a state-of-the-art atmospheric model run with high vertical resolution that is forced solely by prescribed stratospheric ozone depletion. The results provide evidence that anthropogenic emissions of ozonedepleting gases have had a distinct impact on climate not only at stratospheric levels but at Earth's surface as well. PMID- 14551434 TI - Control of crystal size and lattice formation by starmaker in otolith biomineralization. AB - The stone-like otoliths from the ears of teleost fishes are involved in balance and hearing and consist of calcium carbonate crystallites embedded in a protein framework. We report that a previously unknown gene, starmaker, is required in zebrafish for otolith morphogenesis. Reduction of starmaker activity by injection of modified antisense oligonucleotides causes a change in the crystal lattice structure and thus a change in otolith morphology. The expression pattern of starmaker, along with the presence of the protein on the growing otolith, suggest that the expression levels of starmaker control the shape of the otoliths. PMID- 14551436 TI - Does rejection hurt? An FMRI study of social exclusion. AB - A neuroimaging study examined the neural correlates of social exclusion and tested the hypothesis that the brain bases of social pain are similar to those of physical pain. Participants were scanned while playing a virtual ball-tossing game in which they were ultimately excluded. Paralleling results from physical pain studies, the anterior cingulate cortex (ACC) was more active during exclusion than during inclusion and correlated positively with self-reported distress. Right ventral prefrontal cortex (RVPFC) was active during exclusion and correlated negatively with self-reported distress. ACC changes mediated the RVPFC distress correlation, suggesting that RVPFC regulates the distress of social exclusion by disrupting ACC activity. PMID- 14551435 TI - A functional link between RuBisCO-like protein of Bacillus and photosynthetic RuBisCO. AB - The genomes of several nonphotosynthetic bacteria, such as Bacillus subtilis, and some Archaea include genes for proteins with sequence homology to the large subunit of ribulose bisphosphate carboxylase/oxygenase (RuBisCO). We found that such a RuBisCO-like protein (RLP) from B. subtilis catalyzed the 2,3-diketo-5 methylthiopentyl-1-phosphate enolase reaction in the methionine salvage pathway. A growth-defective mutant, in which the gene for this RLP had been disrupted, was rescued by the gene for RuBisCOfrom the photosynthetic bacterium Rhodospirillum rubrum. Thus, the photosynthetic RuBisCO from R. rubrum retains the ability to function in the methionine salvage pathway in B. subtilis. PMID- 14551437 TI - A Neurexin-related protein, BAM-2, terminates axonal branches in C. elegans. AB - Neuronal axons connect to multiple target cells through the formation of collateral branches, but the mechanisms that regulate this process are largely unknown. We show that BAM-2, a neurexin-related transmembrane protein, is required for development of VC motoneuron branches in the worm Caenorhabditis elegans. Expression analysis and ectopic expression experiments suggest that BAM 2 functions as a branch termination cue and reveal a mechanism for selective control of branches that sprout off a primary axon. PMID- 14551439 TI - Imaging of quantum array structures with coherent and partially coherent diffraction. AB - Recent achievements in experimental and computational methods open the possibility of measuring and inverting the diffraction pattern from a finite object of submicrometer size. In this paper the possibilities of such experiments for two-dimensional arrays of quantum dots are discussed. The diffraction pattern corresponding to coherent and partial coherent illumination of a sample was generated. Test calculations based on the iterative algorithms were applied to reconstruct the shape of the individual islands in such a quantum structure directly from its diffraction pattern. It is demonstrated that, in the case of coherent illumination, the correct shape and orientation of an individual island can be obtained. In the case of partially coherent illumination, the correct shape of the particle can be obtained only when the coherence of the incoming beam is reduced to match the size of the island. PMID- 14551438 TI - Gene expression profiles in the brain predict behavior in individual honey bees. AB - We show that the age-related transition by adult honey bees from hive work to foraging is associated with changes in messenger RNA abundance in the brain for 39% of approximately 5500 genes tested. This result, discovered using a highly replicated experimental design involving 72 microarrays, demonstrates more extensive genomic plasticity in the adult brain than has yet been shown. Experimental manipulations that uncouple behavior and age revealed that messenger RNA changes were primarily associated with behavior. Individual brain messenger RNA profiles correctly predicted the behavior of 57 out of 60 bees, indicating a robust association between brain gene expression in the individual and naturally occurring behavioral plasticity. PMID- 14551440 TI - Identification of dislocations in large tetragonal hen egg-white lysozyme crystals by synchrotron white-beam topography. AB - Large tetragonal hen egg-white (HEW) lysozyme crystals have been grown by a salt concentration-gradient method. The grown crystals, of thicknesses greater than 1.5 mm, were observed by means of X-ray topography using white-beam synchrotron radiation. Line contrasts clearly appeared on the Laue topographs. Extinction of the line images was observed in specific reflections. These results mean that the observed lines correspond to dislocation images. From the extinction criterion it is shown that the predominant dislocations are of screw character with <110> Burgers vectors. In addition, dislocation loops with [001] Burgers vectors have been found in a tetragonal HEW lysozyme crystal including some cracks. These results are discussed in the light of dislocation elastic energy and slip systems in the crystals. PMID- 14551441 TI - First experiments on diffraction-enhanced imaging at LNLS. AB - Diffraction-enhanced images have been obtained using two silicon crystals in a non-dispersive set-up at the XRD2 beamline at the Brazilian Synchrotron Light Laboratory (LNLS). A first asymmetrically cut silicon crystal using the (333) reflection vertically expanded the monochromated beam from 1 mm to 20 mm allowing the imaging of the whole sample without movements. A symmetrically cut Si(333) second crystal was used as a Bragg analyzer. Images of biological samples including human tissue were recorded using a direct-conversion CCD detector resulting in enhancement of the contrast compared with absorption-contrast images. PMID- 14551442 TI - High-resolution and high-intensity powder diffractometer at BL15XU in SPring-8. AB - A new ultra-high-resolution powder diffractometer for synchrotron radiation has been constructed at beamline BL15XU, SPring-8. The two-axis diffractometer is optimized for high-flux and high-coherent X-ray beams, which are provided by combining a planar undulator and a large offset rotated-inclined Si(111) double crystal monochromator. The optics design of the diffractometer is based on transmission geometry, which employs a capillary specimen and reflection geometries using a flat-plate specimen. The intensity data are collected using a 2theta step-scan technique in both geometries. The diffractometer can be arranged in a variety of optical configurations, e.g. simple receiving slits, flat crystal analyzer of Ge(111) or Si(111), and in-vacuum-type long horizontal parallel slits. A minimum full width at half-maximum against 2theta was 0.00572 degrees at lambda = 0.63582 A for the (200) reflections from Si powder in the transmission geometry employing the Ge(111) crystal analyzer. A wide temperature range (32-900 K), which is controlled by a He/N(2) gas stream system, is available. 288 structure parameters of a zeolite ZSM-5 sample have been demonstrated to successfully refine with a R(wp) value of 6.96% by a Rietveld analysis of the high-resolution powder diffraction data from a 1 mm-diameter capillary specimen. PMID- 14551443 TI - High-resolution X-ray diffraction beamline at the LNLS for the study of charge, orbital and magnetic structures. AB - A high-resolution X-ray diffraction beamline at the Brazilian Synchrotron Light Laboratory (LNLS) has been commissioned for the study of crystalline magnetic materials. The beamline optics is based on a Rh-coated vertical-focusing X-ray mirror and a sagittal-focusing double-crystal monochromator. The primary instrument is a six-circle diffractometer equipped with energy and polarization analysers and a closed-cycle He cryostat. The beamline source is a bending magnet of the 1.37 GeV storage ring of the LNLS, delivering approximately 4 x 10(10) photons s(-1) at 8 keV at the sample position. Resonant and non-resonant scattering are the main techniques used to study charge, orbital and magnetic structures. Examples of magnetic scattering in Ho and NiO single crystals, as well as orbital ordering in manganites thin films, are presented. PMID- 14551444 TI - Diffraction anomalous fine-structure spectroscopy at beamline BM2 at the European Synchrotron Radiation Facility. AB - Diffraction anomalous fine-structure (DAFS) spectroscopy uses resonant elastic X rays scattering as an atomic, shell and site-selective probe that provides information on the electronic structure and the local atomic environment as well as on the long-range-ordered crystallographic structure. A DAFS experiment consists of measuring the Bragg peak intensities as a function of the energy of the incoming X-ray beam. The French CRG (Collaborative Research Group) beamline BM2-D2AM (Diffraction Diffusion Anomale Multi-longueurs d'Onde) at the ESRF (European Synchrotron Radiation Facility) has developed a state-of-the-art energy scan diffraction set-up. In this article the requirements for obtaining reliable DAFS data are presented and recent technical achievements are reported. PMID- 14551445 TI - Application of a neural network in high-throughput protein crystallography. AB - High-throughput protein crystallography requires the automation of multiple steps used in the protein structure determination. One crucial step is to find and monitor the crystal quality on the basis of its diffraction pattern. It is often time-consuming to scan protein crystals when selecting a good candidate for exposure. The use of neural networks for this purpose is explored. A dynamic neural network algorithm to achieve a fast convergence and high-speed image recognition has been developed. On the test set a 96% success rate in identifying properly the quality of the crystal has been achieved. PMID- 14551446 TI - Development of a novel aspherical mirror bender for an active grating. AB - A novel monolithic mechanical bender has been designed and fabricated to meet the requirements of an active polynomial grating in a new soft X-ray scattering and emission beamline at the National Synchrotron Radiation Research Center, Taiwan. This compact bender achieves nearly fixed center point under different bending conditions. Moreover, the compact bender can be bent to a desirable third-order polynomial surface profile to cancel the defocus and coma aberrations using two PZT actuators. Theoretical analysis reveals that the grating has unprecedented spectral resolving power. A detailed mechanical analysis has been conducted and a prototype bender was fabricated and tested. The results indicate that the performance of the bender is excellent and is therefore suitable to be used in the active grating. PMID- 14551447 TI - A gas microstrip detector for XAS studies in the photon energy region 300-1500 eV. AB - The ability to perform X-ray absorption spectroscopy (XAS) in the 300-1500 eV energy range allows measurements to be made on transition metal compounds. This paper describes a detector and the technique used to perform fluorescent measurements on such materials. A variety of test sample results are shown to illustrate the low energy and energy-resolving capabilities of the detector (based on gas microstrip technology). Two possible applications are also demonstrated. The first shows how the detector can be used to gather X-ray absorption spectra for the L edges of transition metals and K edges of light elements (C, O and N). The other shows how the magnetic immunity of the detector can be exploited to study the magnetic properties of materials. PMID- 14551448 TI - The Mobile Luminescence End-Station, MoLES: a new public facility at Daresbury Synchrotron. AB - A new mobile end-station is described for use on multiple beamlines at the Daresbury synchrotron radiation source (overall excitation range 5 eV to 70 keV) that allows for the detection and dispersion of photoluminescence from solid state samples in the emission range 190-1000 nm (1.2-6.5 eV). The system is fully self-contained and includes sample-cooling facilities for the temperature range 8 330 K using a closed-cycle refrigerator, thus eliminating the need for liquid cryogens. The system also includes solid-state laser sources for use with a variety of pump-probe-type experiments, and an Ar(+) surface-cleaning facility. In order to demonstrate the various capabilities of the system, the results of a variety of experiments are summarized, carried out over the excitation range 5 5000 eV on beamlines 3.2, MPW6.1 and 4.2. These include the optical detection of XAS of L-edge structure in natural minerals and archaeological ceramics, band-gap determinations of wide-band-gap silicates, and pump-probe studies of quartz. PMID- 14551452 TI - A vision of children's dental care. PMID- 14551453 TI - The Mini-Distalizing Appliance: the third dimension in maxillary expansion. PMID- 14551454 TI - Treatment of a unilateral Class II, division 2 malocclusion with a palatally impacted canine. PMID- 14551455 TI - Practice acquisition: financing and related issues. PMID- 14551456 TI - Use of a modified butterfly expander to increase anterior arch length. PMID- 14551457 TI - A preadjusted edgewise approach to cases with anterior crowding. PMID- 14551458 TI - Stimulating tooth eruption with an eruption plate. PMID- 14551459 TI - Reproductive performance following midtrimester termination of pregnancy. AB - The aim of the study was to determine the impact of midtrimester abortion with extra-amniotic PGE(2) on future fertility and reproduction. Two hundred and fifteen women were enrolled. The mean induction-to-abortion interval was 21.3 h. The complication rate was 11.7%. Twenty women (9.3%) were lost to follow-up, 82 patients (38.1%) used contraception. Spontaneous pregnancy occurred in 110 out of 113 women who desired fertility (97%). Three patients were defined as infertile but conceived following treatment. The average time between the abortion and the next conception was 15.9 months (range 1-77 months). Seventy patients (63.7%) delivered at term. Premature delivery occurred in 18 patients (16.4%). Eighteen women aborted spontaneously in the first and 3 in the second trimester. One extrauterine pregnancy occurred. Second-trimester abortion with extra-amniotic PGE(2) is a safe procedure with a low rate of complication. Apparently it does not affect future fertility, but may be associated with an increased rate of premature deliveries in future gestations. PMID- 14551460 TI - Antimicrobial activity of human follicular fluids. AB - The aim of this study was to explore the antimicrobial activity of human follicular fluid (HFF), to test the hypothesis that different strains of the same bacterial species could display different patterns of susceptibility to antimicrobial action of HFF, and to preliminarily investigate the possible mechanism of antimicrobial action of this fluid. Antimicrobial activity of 60 samples of HFF toward 30 Streptococcus agalactiae strains was determined by the agar diffusion method and broth dilution method. To explore the mechanism of antimicrobial activity, biochemical analyses were performed with selected fluid samples. The obtained results indicate that 38.3% fluid samples did not inhibit bacterial growth, 53.3% showed moderate and 8.3% high antimicrobial activity. The tested effect of HFF on S. agalactiae strains was bactericidal and was not strain dependent. Lysozyme activity was detected in HFF exhibiting antimicrobial activity. There were no statistically significant differences in concentrations of estradiol, progesterone, transferrin, iron, total protein and albumin levels among tested samples regardless of the different rate of antimicrobial activity. The obtained results indicate that lysozyme is most probably a crucial antibacterial agent in this fluid; however, some other still unidentified factors may contribute to it. PMID- 14551461 TI - Management of secondary hyperparathyroidism: the importance and the challenge of controlling parathyroid hormone levels without elevating calcium, phosphorus, and calcium-phosphorus product. AB - Secondary hyperparathyroidism (HPT) is a common complication of chronic kidney disease (CKD) that can lead to clinically significant bone disease. Additional consequences of secondary HPT, such as soft-tissue and vascular calcification, cardiovascular disease, and calcific uremic arteriolopathy, may contribute to the increased risk of cardiovascular morbidity and mortality among CKD patients. Secondary HPT arises from disturbances in calcium, phosphorus, vitamin D and parathyroid hormone metabolism, which develop early in the course of CKD and become more prominent as kidney function declines. The standard therapies currently recommended to correct mineral metabolism and bone disease in these patients include calcium supplementation, dietary phosphorus restriction, phosphate-binding agents, and treatment with vitamin D sterols. However, such medications often have significant effects on the serum levels of calcium and phosphorus, which result in exacerbation of the disease and significant extraskeletal morbidity and mortality. Thus, there is a need to identify more effective treatment approaches. This review discusses the pathophysiology of secondary HPT, the challenges faced in the management of this disorder, and the impact of current treatment options on patients' risks of morbidity and mortality. In addition, the development of new, more physiologically relevant therapies, which may lead to successful management of secondary HPT, is reviewed. PMID- 14551462 TI - Activation of protein kinase C isozymes protects LLCPK1 cells from H2O2 induced necrotic cell death. AB - BACKGROUND/AIMS: We have previously reported that ischemia/reperfusion injury (IRI) to the kidney leads to induced expression of RACK1 and changes in the level of expression and subcellular distribution of PKC isozymes alpha, betaII and zeta. In order to further define the role of PKC isozymes in IRI we investigated the effect of activation or inhibition of the isozymes on cytotoxicity mediated by H(2)O(2) in LLCPK(1) cells. METHODS: Cytotoxicity was analyzed by Trypan blue assay and LDH release assay. Translocation of PKC isozymes postinjury in LLCPK1 cells was analyzed by immunostaining and Western blot analysis. RESULTS: Western blot analysis showed that the expression of PKC-alpha was up-regulated in a triphasic pattern with the initial induction within the first 10 min of injury followed by higher levels of expression at 2 and 24 h postinjury. The expression of PKC-zeta was highly induced within the first 15 min of injury but its expression was down-regulated to that of normal levels by 30 min postinjury. Immunocytochemistry showed that both PKC-alpha and PKC-zeta translocated to the nucleus and perinuclear region during H(2)O(2) treatment. Following injury, PKC alpha expression was localized to the nuclear membrane at earlier time points but a translocation to the nucleus occurred at later time points. PKC-zeta translocated to nucleus at 30 minutes post injury and relocated back to the nuclear membrane at later time points. CONCLUSION: These data suggest that activation of PKC-alpha and PKC-zeta is involved in the H(2)O(2) induced injury of LLCPK1 cells. PMID- 14551463 TI - Hyper-vascular change and formation of advanced glycation endproducts in the peritoneum caused by methylglyoxal and the effect of an anti-oxidant, sodium sulfite. AB - OBJECTIVE: Methylglyoxal (MGO) in a heat-sterilized conventional PD solution may damage peritoneal cells directly and/or indirectly by producing advanced glycation endproducts (AGEs). This study was conducted to (a) examine the acute effect of MGO on the peritoneum (including AGE formation) and (b) study the possible AGE suppressive effect of an anti-oxidant, sodium sulfite. METHOD: (1) Human serum albumin (HAS) was continuously incubated with MGO (50 mM) at 37 degrees C for as long as 14 days and the fluorescence intensity (FI) was determined (em. 440, ex. 370). (2) Three types of test solutions - (i) saline; (ii) MGO (20 mM), and (iii) MGO with sodium sulfite (30 mM) - were administered intraperitoneally to 8-week-old rats once a day for 5 consecutive days. The parietal peritoneum was examined macroscopically on the 6th day for immunostaining of anti-AGE antibodies. RESULT: (1) An increase in FI of HSA was observed as a function of the incubation period in the MGO solution. (2) Prominent hypervascularity and intense immunostaining of anti-AGE Ab were noted in MGO-treated rats, whereas the macroscopic alterations were suppressed in the rats that had been treated with sodium sulfite. CONCLUSION: MGO-induced hypervascularity and AGE formation in the peritoneum, as well as macroscopic alterations were suppressed by sodium sulfite. This may indicate that there is a risk of MGO causing a peritoneal injury and that the therapeutic potential of an anti-oxidant for this type of injury may exist. PMID- 14551464 TI - Researching shared governance: a futility of focus. PMID- 14551465 TI - Measuring the impact of collaborative governance: beyond empowerment. PMID- 14551466 TI - The use of a strategic interviewing technique to select the nurse manager. PMID- 14551467 TI - Do no harm: provider perceptions of patient safety. PMID- 14551468 TI - Improving our image a nurse at a time. PMID- 14551470 TI - Beyond the Magnet award: The ANCC Magnet Program as the framework for culture change. AB - The nursing shortage has turned the national spotlight onto the need to improve the conditions under which nurses work. The American Nurses Credentialing Center Magnet Recognition Program offers an evidence-based model for nurse leaders interested in transforming the practice climate. The author describes a community hospital's experience in using the magnet standards as the framework for change. The author describes 5 successful standard-driven strategies that have strengthened department infrastructure and refined processes for retaining a professional nurse workforce. PMID- 14551469 TI - The Vanderbilt Professional Nursing Practice Program, part 2: Integrating a professional advancement and performance evaluation system. AB - Developing a performance-based advancement system requires evaluation tools that capture essential behaviors and outcomes reflective of key nursing functions. To ensure relevance to clinical practice and enhance buy-in from nursing staff, the behaviors and outcomes were defined by a broad cross-section of nursing staff and administrators. The first article (September 2003) in this 3-part series described the foundation for and the philosophical background of the Vanderbilt Professional Nursing Practice Program (VPNPP), the career advancement program under way at Vanderbilt University Medical Center. This second article describes the development of the evaluation tools used in the VPNPP, the implementation and management of this new system, program evaluation, and improvements since the inception of the program. Additionally, the authors present the challenges and lessons we learned in development and implementation of a dynamic evaluation system that supports our career advancement program. The process of advancing within the program will be described in part 3. PMID- 14551471 TI - Ensuring success: research by committee. AB - The Nursing Research Committee at a community teaching hospital conducted a research study on alternatives to restraints in a confused population. The authors describe the process this committee used to successfully conduct the study, from starting a research focus group and setting goals to choosing a research topic, designing the study, and collecting data. Critical success factors for completing research by committee are described. PMID- 14551472 TI - Nursing workforce management: using benchmarking for planning and outcomes monitoring. AB - The goal of maintaining the balance between demand for nursing care and the supply of nurses is the basis for state and regional planning to meet future nursing workforce needs. Projecting nursing supply needs solely on the basis of historical registered nurse to population ratios does not specifically consider the healthcare needs of the population. The authors present a model in which state population data and nursing resource data are compared with national data to assist state planning groups in developing an effective workforce management plan. Integrating population demographic data, healthcare needs of citizens, and nurse resource factors at the state level can improve the accuracy of projection statistics. PMID- 14551473 TI - Designing a leadership development program for nurse managers: an evidence-driven approach. AB - The authors describe an evidence-driven approach used in designing a leadership development program targeted for nurses in middle management positions in an academically affiliated integrated healthcare system. A qualitative study was conducted to investigate leadership development needs and experiences of nurse managers across the health system. Study findings were used to inform educational programs and organizational initiatives responsive to needs of both novice and expert nurse managers across diverse clinical settings. PMID- 14551475 TI - Incidence of influenza in Finnish children. AB - BACKGROUND: Influenza is an important cause of respiratory illness in children, but data on virologically confirmed influenza infections in children treated as outpatients are limited. METHODS: We carried out a prospective cohort study of normal children younger than 13 years (n = 1338) in the winter of 2000 to 2001. During the study period of 32 weeks, the children were examined at the study clinic whenever they had fever or signs of respiratory infection. Nasal swabs were obtained during each episode of infection for determination of the viral etiology of the illness. RESULTS: The overall attack rate of influenza in the cohort was 18.8%. Influenza viruses were isolated from the children from the beginning of November 2000 through May 2001. Virtually in each week between mid November and the end of April (a period of 24 weeks), influenza viruses accounted for at least 5% of all respiratory infections in the children. During the peak of the epidemic, the percentage of influenza-positive children exceeded 20%. CONCLUSIONS: This study confirms the important role of influenza as a cause of acute respiratory infections in children, even in winters of mild or moderate influenza activity. The study also shows that influenza viruses may circulate in the community at substantial levels much longer than previously thought. PMID- 14551476 TI - Socioeconomic impact of influenza on healthy children and their families. AB - BACKGROUND: Recent studies indicate that influenza can be clinically important in otherwise healthy children. However, the interpretation of many studies is limited because of lack of laboratory confirmation of influenza-like illnesses. Therefore it is difficult to conclude whether the socioeconomic impact of influenza justifies vaccinating all children regardless of age or underlying chronic disorders. METHODS: We prospectively collected data from 3771 children younger than 14 years of age presenting to emergency departments or primary care pediatricians with symptoms of respiratory tract infection during the influenza season of 2001 to 2002. Influenza infections were verified by virus culture or polymerase chain reaction. We additionally randomized 303 children age 6 months to 5 years to receive either influenza vaccine (n = 202) or no vaccination (n = 101) before the influenza season. The socioeconomic impact of influenza was assessed for both the participating children and their household contacts. RESULTS: Influenza was documented in 352 (9.3%) of the 3771 children. Compared with influenza-negative children, children with influenza had longer durations of fever and absenteeism from day care or school (P < 0.0001). Further the numbers of medical visits, missed work or school days and the need for help at home to care for the sick children were higher among the household contacts of influenza positive children (P < 0.0001). Influenza vaccination reduced significantly the direct and indirect influenza-related costs in healthy children and their unvaccinated family members. CONCLUSIONS: The findings of this study support a wider use of influenza vaccine in healthy children of all ages to reduce the socioeconomic burden of influenza on the community. PMID- 14551477 TI - Burden of influenza in children: preliminary data from a pilot survey network on community diseases. AB - BACKGROUND: The burden of influenza and its clinical presentation are both poorly documented in French children. METHODS: To measure the prevalence of acute respiratory infections over two winters (2000 to 2001 and 2001 to 2002), we performed a prospective community survey of 0- to 15-year-olds (n = 11 500 and 40 000, respectively) through a network [Grippe et Infections Respiratoires Aigues Pediatriques (GIRAP)] of general practitioners and pediatricians. Influenza viruses were detected by antigen detection and culture from nasopharyngeal swabs. Data on the clinical presentation of children infected with influenza A/H1N1, A/H3N2 or B viruses were derived from standardized forms. RESULTS: The prevalences of symptomatic acute respiratory infections were 28% and 27% for 2000 to 2001 and 2001 to 2002, respectively. In the winter of 2000 to 2001, a mild outbreak of influenza A/H1N1 affected 5.4% of the study population. The attack rates of influenza A/H1N1 were highest in children 5 to 15 years of age. In the winter of 2001 to 2002, influenza activity remained at a subepidemic level (5.9%; A/H3N2, 60%; B, 40%). The incidence of influenza A/H3N2 infections was highest in 2- to 4-year-old children, whereas influenza B strains were most frequently detected in children 4 to 10 years of age. CONCLUSIONS: Despite subepidemic levels of influenza, the GIRAP network was able to detect influenza A or B viruses in 35 to 60% of the samples during the peak of influenza epidemic. The frequency of influenza virus excretion in children with mild clinical symptoms indicates that children are an important reservoir and source of transmission of this virus. PMID- 14551478 TI - Influenza in children: the German perspective. AB - BACKGROUND: Influenza is recognized as an important disease in infants and children, but epidemiologic data on influenza in children are limited. METHODS: We analyzed German epidemiologic data from three sources: a population-based study in children 0 to 3 years of age [Paediatric Respiratory Infection in Germany (PRIDE) study]; a multicenter hospital-based survey of children 0 to 16 years of age (www.pid-ari.net); and a sentinel study (Deutsche Arbeitsgemeinschaft Influenza) comparing influenza activity and numbers of consultations for respiratory infections in children and adults. RESULTS: In 1996 to 2001, the PID-ARI-net and the PRIDE studies showed that the rate of influenza associated hospitalizations was approximately 120/100 000 children (0 to 5 and 0 to 3 years of age, respectively). Children born prematurely and those with underlying cardiac or pulmonary disease were at increased risk of hospitalization. The mean duration of hospital stay was 6.3 days, which is equal to the mean length of hospitalization for respiratory syncytial virus infections in continental Europe. Few nosocomial infections were reported. The outpatient consultation rate per year for lower respiratory tract illness caused by influenza virus was 1.1/100 children. CONCLUSION: Direct and indirect epidemiologic evidence indicates that influenza is a significant health hazard for all children. PMID- 14551480 TI - Influenza A community-acquired pneumonia in East London infants and young children. AB - BACKGROUND: Community-acquired pneumonia (CAP) is common in young children, but there are few data in Europe on influenza A virus as a cause of childhood CAP. The aim of this study was to determine the relative contributions of different etiologic agents to CAP in children. METHODS: This was a 6-month prospective study of pediatric accident and emergency and general practice consultations with a diagnosis of CAP. Nasopharyngeal aspirates for viral immunofluorescence and PCR studies and blood cultures for bacterial studies were taken from 51 children with symptoms, signs and chest radiographic features that satisfied a diagnosis of pneumonia. RESULTS: An etiologic agent was isolated from 25 patients (49%). A viral cause was identified in 22 patients (43%), and influenza A virus and respiratory syncytial virus (RSV) were detected in 16 and 18% of all cases, respectively. Only four patients (8%) had a positive bacterial blood culture; three had Streptococcus pneumoniae and one had Neisseria meningitidis W135. Mycoplasma pneumoniae was detected in 2 children, and mixed infections were detected in 5 (10%). The use of viral PCR increased the detection rate of influenza A virus by 100%. CONCLUSION: Influenza A virus caused more than one third of all viral CAP cases, a rate comparable with that of RSV CAP. Viral PCR doubled the diagnostic yield of influenza A virus. The clinical burden of influenza A CAP was comparable with that of RSV CAP, as measured by the duration of fever, hospital stay and total duration of illness. PMID- 14551479 TI - Influenza burden in children newborn to eleven months of age in a pediatric emergency department during the peak of an influenza epidemic. AB - BACKGROUND: The aim of this study was to determine the burden of influenza related diseases in children 0 to 11 months of age during the peak of the 2001 to 2002 influenza epidemic. METHODS: This was a prospective study at the Pediatric Emergency Department of Edouard Herriot tertiary teaching hospital in Lyon, France. The study included 304 infants 0 to 11 months of age. Consecutive patients were systematically enrolled during the 4 weeks of the influenza epidemic peak (Weeks 3 to 6, 2002). Influenza viruses were detected by antigen detection and virus culture from nasal swabs. Structured telephone interviews were conducted on Days 8 and 15 after virus detection. There was also a 6-month survey into the medicoadministrative database to detect late complications that required delayed hospitalization of influenza-positive children. RESULTS: Influenza virus was detected in 99 (33%) of 304 patients (A/H3N2 in 30% and B in 3%). Nonrespiratory symptoms were the dominant clinical manifestations in 30% of influenza-positive children. One child with influenza presented with febrile seizures. Twenty (20%) children with influenza were hospitalized. Parents reported recovery from the illness in 63 and 94% of children on Days 8 and 15, respectively. The median length of an influenza episode was 8 days. CONCLUSIONS: Our results confirm the high prevalence of influenza-related morbidity in infants during the epidemic peak. One child in three consulting to the pediatric emergency room had a virologically confirmed influenza infection regardless of the body temperature. Every fifth child with influenza was admitted to hospital, which corresponds to an admission rate of 237 per 100 000 children 0 to 11 months of age. PMID- 14551481 TI - Influenza-related hospitalizations among young children in Leicestershire. AB - BACKGROUND: Population-based data on influenza hospitalizations are unavailable in the United Kingdom, but they represent an essential component of health economic analyses that could support the use of vaccines and antiinfluenza drugs in healthy children. We collected data on hospitalizations for influenza infections among young children in Leicester, UK. METHODS: This prospective, longitudinal, noninterventional single center study was conducted at the Children's Hospital in Leicester, which provides inpatient pediatric care to a total population of approximately 1 million. We studied children <6 years of age between October 14, 2001, and June 30, 2002, who were admitted to the hospital with an acute respiratory tract illness, seizures, specified acute febrile gastrointestinal illness or any acute febrile illness or apnea or other life threatening events in infants <12 months of age. Nasopharyngeal swabs obtained within 24 h of hospital admission were examined for influenza, respiratory syncytial virus and human metapneumovirus by PCR. RESULTS: Of 7165 clinical episodes that were assessed in the Children's Hospital between October 1 and June 30, 2441 (34.1%) were caused by acute respiratory illness. Overall 33 (5.4%) of 613 children analyzed had an influenza A or B virus infection, including 19 (5.0%) of 381 children with acute respiratory illness and 14 (6.0%) of the remaining 232 children. CONCLUSIONS: Influenza is evidently an important cause of hospitalization among young children, even during limited outbreaks of influenza. Further analyses will enable us to estimate age-related admission rates for influenza and to compare the burden from influenza with that for respiratory syncytial virus and human metapneumovirus. PMID- 14551482 TI - Impact of influenza on young children and the shaping of United States influenza vaccine policy. AB - BACKGROUND: In 2002 the Advisory Committee on Immunization Practices (ACIP) began encouraging annual influenza vaccination of children 6 to 23 months of age, when feasible. METHODS: Literature and issues related to annual influenza vaccination of young children were reviewed. RESULTS: The ACIP first encouraged influenza vaccination of children 6 to 23 months of age in 2002 because recent studies showed that influenza-related hospital admissions were substantially higher among healthy children <2 years than among healthy older children or young adults. However, the ACIP deferred a full recommendation for several reasons, including limited safety and efficacy data on trivalent inactivated influenza virus in the 6- to 23-month age group, the need for more education of parents and physicians and concerns over the stability and adequacy of the vaccine supply. CONCLUSIONS: The risk of hospital admission from influenza-related causes is high in young children and similar to the risk in the elderly and other high risk groups for whom annual influenza vaccination is already recommended. Data from additional studies, especially those on vaccine safety and efficacy, will be important for proceeding to a full recommendation for annual influenza vaccination of children 6 to 23 months. PMID- 14551484 TI - Substantial variability in community respiratory syncytial virus season timing. AB - BACKGROUND: Respiratory syncytial virus (RSV) is the major cause of bronchiolitis and pneumonia in young children. Prevention of RSV disease in children in certain high risk groups through use of immunoglobulin preparations has been recommended by the American Academy of Pediatrics since 1998. A more precise understanding of the timing of annual RSV epidemics should assist providers in maximizing the benefit of these preventive therapies. The objective of this study was to determine whether current national RSV surveillance data could be used to define the timing of seasonal outbreaks METHODS: Weekly RSV testing data from the National Respiratory and Enteric Viruses Surveillance System for the period July 1990 through June 2000 were analyzed. RSV season onset week, peak week and duration were calculated for the entire United States, Census regions and select local laboratories. Season variability was estimated by comparing calculations for individual RSV seasons to median measurements for the entire surveillance period RESULTS: RSV seasons in the South region began significantly earlier (P < 0.05) and lasted longer (P < 0.05) than seasons for the rest of the nation. RSV seasons in the Midwest region began significantly later (P < 0.01) and were shorter (P < 0.05) than those for the rest of the nation. Local RSV seasons varied substantially by year and by laboratory. The variability between laboratories generally increased with distance between laboratories CONCLUSIONS: Onset weeks and durations of RSV seasons vary substantially by year and location. Local RSV data are needed to accurately define the onset and offset of RSV seasons and to refine timing of passive immune prophylaxis therapy recommendations. PMID- 14551483 TI - Impact of intrapartum antibiotics on the care and evaluation of the neonate. AB - BACKGROUND: Management of infants whose mothers receive intrapartum antibiotic prophylaxis (IAP) is controversial. In 1996 consensus guidelines for prevention of neonatal Group B streptococcal disease included an algorithm for management of infants whose mothers received IAP. To assess practices for testing and treatment of infants, we evaluated a population-based sample of deliveries to see whether excessive evaluation and treatment occurs after IAP. METHODS: Medical records for 869 deliveries in Connecticut during 1996 were sampled. IAP was administered in 96 full term deliveries. We excluded infants <37 weeks and those with intrapartum fever. We reviewed hospital records for infants born after IAP (n = 81) and a random sample of those not exposed (n = 180). Analyses were conducted with sample weights to account for unequal probability of selection. RESULTS: Infants whose mothers received IAP were more likely to have complete blood counts, (26% vs. 9% P = 0.05) but were no more likely to receive antibiotics in the first week of life (P = 0.48), have an intravenous catheter placed (P = 0.83), or to have other invasive procedures. Mean length of hospital stay was 6 h longer for infants born by vaginal delivery to mothers who had IAP (47.0 h) than for those without IAP (41.3 h) (P = 0.06). CONCLUSION: Despite concerns that IAP guidelines would result in excessive neonatal evaluations, infants sampled whose mothers received IAP were not more likely to undergo invasive procedures or to receive antibiotics. Consistent with the guidelines, collection of complete blood counts was more common among such infants. PMID- 14551485 TI - Impact of highly active antiretroviral therapy on the morbidity and mortality in Spanish human immunodeficiency virus-infected children. AB - BACKGROUND: Highly active antiretroviral therapy (HAART) slows the progression of HIV disease and lowers mortality and morbidity in adults. The impact on the disease course in children has not been still completely elucidated. Furthermore the effect of HAART on organ-specific complications of HIV is unknown. OBJECTIVES: To assess the effect of HAART on the progression of HIV infection, mortality, organ-specific complications, number of infections and hospitalizations in HIV-1-infected children. PATIENTS AND METHODS: Records of HIV 1-infected children were reviewed in a large referral pediatric hospital. Patients were divided into three groups: children who did not receive antiretroviral therapy (Group 1); children who received mono- or bitherapy (Group 2); and patients who received HAART (Group 3). Endpoints analyzed were progression to AIDS and mortality among AIDS patients and overall. RESULTS: One hundred seven children have been evaluated. Actuarial survival at 5 years of age was 33% in Groups 1 and 2 compared with 100% in Group 3 (P < 0.01). At 5 years of age, the proportion of children progressing to AIDS was 76% in Groups 1 and 2, compared with 26% in Group 3 (P < 0.01). At 5 years of follow-up, there were 45 cases of organ-specific complications in patients without HAART. No children without organ-specific complications when HAART was started have developed them after 5 years (P < 0.01). In patients without HAART there were 9 cases of lymphoid interstitial pneumonia, and there was none in Group 3 (P < 0.01). The incidence rates of infections and hospitalizations were 2.83 and 0.52 per patient year, respectively, in children who did not receive HAART and 0.75 and 0.17 when they were managed with HAART (relative risk, 0.26 and 0.32). CONCLUSIONS: HAART is associated with a marked decline in the progression to AIDS, improved survival in HIV-1-infected children, reduced incidence of infections and hospitalizations and decreased incidence of some organ-specific complications of HIV. PMID- 14551486 TI - Age dependence of in vitro survival of meningococci in whole blood during childhood. AB - OBJECTIVES: To determine the association between the ability of a different strains of meningococci to survive in whole blood and the age of the donor. METHODS: A panel of serogroup B and a serogroup C strain of Neisseria meningitidis was tested in an ex vivo whole blood model. Blood from 81 healthy children and 20 adults and from children during convalescence from serogroup B (55 patients) or serogroup C (43 patients) meningococcal infection was assessed. RESULTS: Age-dependent acquisition of whole blood killing of serogroup B and C bacterial isolates was demonstrated in healthy children with an inverse relationship to the reported incidence of disease. After infection with serogroup B or C meningococci, evidence of whole blood killing of the bacteria was found even in blood from children <2 years of age, the survival of a serogroup B strain, MC58, being reduced compared with that in healthy children (median, 64% compared with 194.5% survival at 90 min). In both affected children and controls, there was a significant correlation between whole blood killing of strain MC58 and of other serogroup B and C meningococci. CONCLUSIONS: The whole blood model measures both humoral and cellular mechanisms responsible for the bactericidal activity of blood. The model was first described 80 years ago, but this is the first description of its age dependency. Acquisition of bactericidal activity was more rapid in children infected and is directed at various strains of meningococci, indicating the presence of a cross-reactive antigen(s). PMID- 14551487 TI - Elimination of acute hepatitis B among adolescents after one decade of an immunization program targeting Grade 6 students. AB - BACKGROUND: British Columbia introduced a preadolescent hepatitis B (HB) immunization program in 1992. This study documents trends in the reported rate of acute HB disease since 1992 and examines factors bearing on the rate of infection throughout the period of program implementation. METHODS: All Grade 6 students were eligible for immunization. Vaccine uptake was reported annually for every school. Acute HB infections were reported by physicians and by biomedical laboratories. Year-to-year trends were analyzed overall and by age group using the electronic public health information system and S-plus. Likelihood ratio tests were used to establish whether a variable was associated with the rate of acute HB in a given cohort. Poisson regression was applied to determine which variables were independently associated with the rate of acute HB. RESULTS: Immunization coverage ranged between 90 and 93% for each year between 1993 and 2001. The overall rate of reported acute HB declined from 7 per 100,000 to just more than 2 per 100,000, whereas that in 12- to 21-year-olds declined from 1.7 to 0 per 100,000 over this one decade period. In the final Poisson regression model, the rate of acute HB infection was significantly associated with year, urban region and lower vaccine uptake. There was an interaction between region and vaccine uptake such that higher vaccine uptake appeared more protective in rural than in urban regions. CONCLUSIONS: Acute HB has been eliminated in the immunized adolescent cohort. A higher carrier rate in urban regions most likely explains the apparent difference in program effectiveness between urban and rural regions. PMID- 14551488 TI - Acute mastoiditis in Southern Israel: a twelve year retrospective study (1990 through 2001). AB - BACKGROUND: Acute mastoiditis is a serious complication of acute otitis media (AOM) and has been increasingly reported in the last decade. OBJECTIVES: To report the experience accumulated with acute mastoiditis at the Soroka University Medical Center, Beer-Sheva, Israel, in a period of increasing antimicrobial resistance with Streptococcus pneumoniae. PATIENTS AND METHODS: We reviewed the records of all children with acute mastoiditis hospitalized from 1990 through 2001. Acute mastoiditis was diagnosed when one or more of the physical signs of mastoiditis (swelling, erythema, tenderness of the retroauricular area and anteroinferior displacement of the auricle) were diagnosed in the presence of concomitant or recent (< or =4 weeks) AOM. RESULTS: One hundred sixteen episodes of acute mastoiditis occurred in 101 children age 2 months to 14 years (median, 25 months; 19% <1 year old). The average yearly incidence was 6.1 cases per 100 000 population <14 years old, with a significant increase in the number of cases during the study period. Acute mastoiditis was the first evidence of AOM in 10 (10%) patients. Fever >38 degrees C and >15,000 WBC/mm3 were present in 67 and 43% of cases, respectively. Irritability, retroauricular swelling, redness and protrusion of the auricle occurred more commonly in patients <3 years old (79, 90, 84 and 76% vs. 28, 42, 45 and 30%, respectively, in patients > or =3 years old; P < 0.002). Computed tomography scans were performed in 54 of 116 (47%) cases and revealed bone destruction in 38 (70%). Periosteal abscess and lateral sinus vein thrombosis were diagnosed in 8 and 2 patients, respectively. Simple mastoidectomy was done in 32 of 116 (28%) cases, after no response to intravenous antibiotics was observed; ventilation tubes were inserted in 12 patients. Cultures were obtained at admission in 83 (72%) episodes (71 by tympanocentesis and 12 from ear discharges). Overall 43 pathogens were isolated: 34 at admission; 14 at surgery; and 5 at both occasions. The most commonly isolated organisms were S. pneumoniae (14 of 43, 33%; 4 penicillin-nonsusceptible), Streptococcus pyogenes (11 of 43, 26%), nontypable Haemophilus influenzae (6 of 43, 14%), Pseudomonas aeruginosa (5 of 43, 12%) and Escherichia coli (4 of 43, 9%). All S. pneumoniae organisms were isolated between 1996 and 2001. CONCLUSIONS: (1) The incidence of acute mastoiditis in children in Southern Israel is greater than that reported in the literature; (2) a significant increase in the number of cases was recorded during the study period; (3) the clinical picture of acute mastoiditis was more severe in infants and young children; (4) the pathogen distribution in acute mastoiditis differs from that of AOM with significantly higher rates of S. pyogenes and lower rates of nontypable H. influenzae recovery; (5) penicillin-nonsusceptible S. pneumoniae played only a minor role in the etiology of acute mastoiditis in Southern Israel. PMID- 14551490 TI - Neonatal enterovirus infections: emphasis on risk factors of severe and fatal infections. AB - OBJECTIVES: Neonatal enterovirus infections have diverse manifestations, from asymptomatic to fatal. An understanding of the risk factors associated with severe cases might help to reduce enterovirus-related morbidity and mortality. METHODS: From July 1989 through June 1998, neonates with virus culture-confirmed nonpolio enterovirus infection at Chang Gung Children's Hospital were enrolled in the study and divided into three groups: nonspecific febrile illness; aseptic meningitis; and hepatic necrosis with coagulopathy (HNC). Demographic factors, clinical manifestations, laboratory data and outcome were analyzed to reveal factors associated with clinical severity and fatality. RESULTS: There were 146 cases including 43 neonates with nonspecific febrile illness, 61 with aseptic meningitis and 42 with HNC. By multiple logistic regression analysis, the most significant factors associated with HNC were prematurity, maternal history of illness, earlier age of onset (< or =7 days), higher white blood cell count (WBC > or =15 000/mm3) and lower hemoglobin (< or =10.7 g/dl). In 10 (24%) of 42 cases, HNC was fatal. In comparison with nonfatal cases of HNC, fatal cases had higher WBC, lower hemoglobin, higher bilirubin and higher incidence of concurrent myocarditis. Multivariate analysis showed the most significant factors associated with fatality from HNC to be total bilirubin >14.3 mg/dl (adjusted odds ratio, 29.1; 95% confidence interval, 2.5 to 355.5; P = 0.007) and concurrent myocarditis (adjusted odds ratio, 13.7; 95% confidence interval, 1.1 to 177.2; P = 0.04). Intravenous immunoglobulin did not correlate with clinical outcomes in cases with HNC. CONCLUSIONS: Prematurity, maternal history of illness, earlier age of onset, higher WBC and lower hemoglobin are significant factors associated with HNC; higher total bilirubin and concurrent myocarditis were most significantly associated with fatality from HNC. PMID- 14551489 TI - Double blind, randomized, placebo-controlled study of dexamethasone therapy for hematogenous septic arthritis in children. AB - BACKGROUND: Septic arthritis is associated with residual dysfunction in 10 to 25% of affected children. Concentrations of cytokines detected in synovial fluid of children with bacterial arthritis correlate with the severity of inflammation. Treatment with dexamethasone decreased cartilage degradation in experimental Haemophilus influenzae b and Staphylococcus aureus arthritis. ENDPOINTS: To decrease the number of patients with residual dysfunction of the affected joint at the end of therapy and at 6 and 12 months and to speed clinical recovery by the administration of dexamethasone. METHODS: In a double blind manner we randomly selected 123 children with suspected hematogenous bacterial arthritis to receive dexamethasone or saline for 4 days. Antibiotic therapy was tailored according to age and the recovered pathogen. RESULTS: Of the 123 children enrolled, 61 were assigned to the dexamethasone group and 62 to the placebo group. Only 50 and 50 patients in each group were evaluable. The 2 groups of patients were comparable with respect to age, sex, duration of symptoms, pathogen, affected joint and therapeutic and diagnostic procedures. Staphylococcus aureus accounted for 67% of the isolates, Haemophilus influenzae type b for 13% and Streptococcus pneumoniae for 9%. Dexamethasone therapy reduced residual dysfunction at the end of therapy, P = 0.000068; at 6 months, P = 0.00007; and at 12 months, P = 0.00053 of follow-up and shortened the duration of symptoms (P = 0.001) during the acute phase. The 26% incidence of residual dysfunction in the control patients was similar to the 25% found in other series. CONCLUSIONS: A short course of dexamethasone reduced residual joint dysfunction and shortened significantly the duration of symptoms in children with documented hematogenous septic arthritis. These results suggest that a 4-day course of low dose dexamethasone given early benefits children with hematogenous septic arthritis. PMID- 14551491 TI - Procalcitonin in pediatric emergency departments for the early diagnosis of invasive bacterial infections in febrile infants: results of a multicenter study and utility of a rapid qualitative test for this marker. AB - BACKGROUND: Procalcitonin (PCT) is a potentially useful marker in pediatric Emergency Departments (ED). The basic objectives of this study were to assess the diagnostic performance of PCT for distinguishing between viral and bacterial infections and for the early detection of invasive bacterial infections in febrile children between 1 and 36 months old comparing it with C-reactive protein (CRP) and to evaluate the utility of a qualitative rapid test for PCT in ED. METHODS: Prospective, observational and multicenter study that included 445 children who were treated for fever in pediatric ED. Quantitative and qualitative plasma values of PCT and CRP were correlated with the final diagnosis. To obtain the qualitative level of PCT the BRAHMS PCT-Q rapid test was used. RESULTS: Mean PCT and CRP values in viral infections were 0.26 ng/ml and 15.5 mg/l, respectively. The area under the curve obtained for PCT in distinguishing between viral and bacterial infections was 0.82 (sensitivity, 65.5%; specificity, 94.3%; optimum cutoff, 0.53 ng/ml), whereas for CRP it was 0.78 (sensitivity, 63.5%; specificity, 84.2%; optimum cutoff, 27.5 mg/l). PCT and CRP values in invasive infections (PCT, 24.3 ng/ml; CRP 96.5 mg/l) were significantly higher than those for noninvasive infections (PCT, 0.32 ng/ml; CRP, 23.4 mg/l). The area under the curve for PCT was 0.95 (sensitivity, 91.3%; specificity, 93.5%; optimum cutoff, 0.59 ng/ml), significantly higher (P < 0.001) than that obtained for CRP (0.81). The optimum cutoff value for CRP was >27.5 mg/l with sensitivity and specificity of 78 and 75%, respectively. In infants in whom the evolution of fever was <12 h (n = 104), the diagnostic performance of PCT was also greater than that of CRP (area under the curve, 0.93 for PCT and 0.69 for CRP; P < 0.001). A good correlation between the quantitative values for PCT and the PCT-Q test was obtained in 87% of cases (kappa index, 0.8). The sensitivity of the PCT-Q test (cutoff >0.5 ng/ml) for detecting invasive infections and differentiating them from noninvasive infections was 90.6%, with a specificity of 83.6%. CONCLUSIONS: PCT offers better specificity than CRP for differentiating between the viral and bacterial etiology of the fever with similar sensitivity. PCT offers better sensibility and specificity than CRP to differentiate between invasive and noninvasive infection. PCT is confirmed as an excellent marker in detecting invasive infections in ED and can even make early detection possible of invasive infections if the evolution of the fever is <12 h. The PCT-Q test has a good correlation with the quantitative values of the marker. PMID- 14551492 TI - Regional variation in the cost effectiveness of childhood hepatitis A immunization. AB - BACKGROUND: Routine childhood hepatitis A immunization is recommended in regions with incidence rates twice the national average, but it may be cost-effective in a wider geographic area. OBJECTIVE: To evaluate the costs and benefits of potential hepatitis A immunization of healthy US children in regions with varying hepatitis A incidences. METHODS: We considered vaccination of the 2000 US birth cohort in states defined by historic hepatitis A incidence rates. Infections among potential vaccinees and their personal contacts were predicted from age 2 through 85 years. Net vaccination costs were estimated from health system and societal perspectives and were compared with life-years saved and quality adjusted life years (QALYs) gained using a 3% discount rate. RESULTS Nationally vaccination would prevent >75 000 cases of overt hepatitis A disease. Approximately two-thirds of health benefits would accrue to personal contacts rather than to vaccinees themselves. In states with incidence rates of > or =200%, 100 to 199%, 50 to 99% and <50% the national average, societal costs per QALY gained would be <0, <0, 13,800 and 63,000 US dollars, respectively. Nationally vaccination would cost 9100 US dollars per QALY gained from the perspective of the health system and 1400 US dollars per QALY gained from society's perspective. Results are most sensitive to vaccination costs and rates of disease transmission through personal contact. CONCLUSION: Childhood hepatitis A vaccination is most cost-effective in areas with the highest incidence rates but would also meet accepted standards of economic efficiency in most of the US. A national immunization policy would prevent substantial morbidity and mortality, with cost effectiveness similar to that of other childhood immunizations. PMID- 14551493 TI - Safety, immunogenicity and efficacy in healthy infants of G1 and G2 human reassortant rotavirus vaccine in a new stabilizer/buffer liquid formulation. AB - BACKGROUND: A refrigerator-stable rotavirus (RV) vaccine that withstands gastric acid is anticipated to permit more widespread use of RV vaccine. OBJECTIVE: We investigated for the first time in infants an oral, liquid formulation of G1 and G2 human bovine reassortant rotavirus vaccine (HRRV) with a new stabilizer/buffer (S/B) containing sucrose, sodium phosphate and sodium citrate. METHODS: During 1997 through 1998, 731 healthy infants approximately 2 to 4 months of age were enrolled at 19 US sites to receive 3 HRRV or placebo doses approximately 6 to 8 weeks apart in a partially double blinded study. Infants were randomized to: (1) HRRV with no S/B but with prefeeding; (2) HRRV plus 1 of 3 different concentrations/volumes of S/B; or (3) placebo. RESULTS: No serious vaccine related adverse experiences or intussusception cases were reported. No statistically significant differences were observed between vaccine and placebo recipients for fever (> or =38.1 degrees C) 0 to 7 days after any dose, irritability, vomiting or diarrhea incidence 0 to 42 days after any dose. Vaccine virus shedding among vaccine recipients was uncommon. Among S/B vaccine groups, proportions of infants with a > or =3-fold titer rise from baseline to Postdose 3 for G1 serum-neutralizing antibody (SNA), G2 SNA, WC3 SNA, serum anti-RV IgA, serum anti-RV IgG and stool anti-RV IgA were generally similar to those of the prefed non-S/B group. CONCLUSIONS: HRRV with a new S/B was generally well tolerated; immunogenicity was generally similar to the prefed non-S/B group. No intussusception cases were reported, but the small sample size precluded a definitive conclusion. A large international clinical study is under way to address safety and efficacy of an S/B formulation of a pentavalent version of HRRV. PMID- 14551494 TI - Lemierre's syndrome. PMID- 14551495 TI - Human metapneumovirus, a newly emerging respiratory virus. PMID- 14551496 TI - Isolation of exfoliative toxin-producing Staphylococcus aureus. PMID- 14551497 TI - Age-related immune reconstitution during highly active antiretroviral therapy in human immunodeficiency virus type 1-infected children. PMID- 14551498 TI - Varicella associated with Staphylococcus aureus purulent pericarditis. PMID- 14551499 TI - Correlation between bacteriologic and clinical endpoints in trials of acute otitis media. PMID- 14551500 TI - Targeting the type 1 insulin-like growth factor receptor as anti-cancer treatment. AB - The type 1 insulin-like growth factor receptor (IGF1R) is overexpressed by many tumors, and mediates growth, motility and protection from apoptosis. Inhibition of IGF1R expression or function has been shown to block tumor growth and metastasis, and enhance sensitivity to cytotoxic drugs and irradiation. Thus the IGF1R is a highly promising anti-cancer treatment target. This review describes approaches to target the IGF1R using antibodies, small molecule inhibitors of the IGF1R tyrosine kinase, and molecular agents such as antisense and small interfering RNAs. Problems for the clinical introduction of this approach may include toxicity due to normal tissue IGF1R expression and cross-reactivity with the insulin receptor. The next few years will see clinical trials of IGF1R targeting, which offers genuine potential to inhibit tumor growth and chemoresistance in patients with cancer. PMID- 14551501 TI - A shikonin derivative, beta-hydroxyisovalerylshikonin, is an ATP-non-competitive inhibitor of protein tyrosine kinases. AB - Studies of the mechanism of action of a shikonin derivative, beta hydroxyisovalerylshikonin (beta-HIVS), have revealed that beta-HIVS inhibits the protein tyrosine kinase (PTK) activities of the receptor for epidermal growth factor and v-Src. In this review, we compare the characteristics of the inhibition of PTK activity by beta-HIVS with those of other inhibitors of PTKs. The chemical structure of beta-HIVS is completely different from that of ATP and it does not resemble any of the PTK inhibitors reported to date, except that it includes the benzylidene moiety. In contrast to most PTK inhibitors, the mechanism of inhibition by beta-HIVS is non-competitive with respect to ATP, but competitive with respect to its peptide substrate. This feature of the mechanism of inhibition of PTK by beta-HIVS suggests that it might be useful in a clinical setting with other PTK inhibitors. When Bcr-Abl-positive, human leukemia K562 cells were treated simultaneously with beta-HIVS and STI571 (Gleevec), these compounds had a synergistic effect on both the induction of apoptosis in K562 cells and the inhibition of the phosphorylation activity of PTK, probably because the mechanism of interference with phosphorylation by beta-HIVS and the binding site of beta-HIVS are different from those of STI571. PMID- 14551502 TI - Dihydropyrimidine dehydrogenase inhibition as a strategy for the oral administration of 5-fluorouracil: utility in the treatment of advanced colorectal cancer. AB - Dihydropyrimidine dehydrogenase (DPD), the rate-limiting enzyme in 5-fluorouracil (5-FU) catabolism, has been a key target for the development of novel oral fluoropyrimidines. DPD-inhibiting oral fluoropyrimidines showing promise in early clinical studies included UFT (the 5-FU prodrug, tegafur, plus the DPD substrate, uracil), eniluracil (an irreversible DPD inhibitor that improves the oral bioavailability of 5-FU) and S-1 (tegafur plus a reversible DPD inhibitor, 5 chloro-2,4-dihydroxypyridine, and oxonic acid). However, results from phase II/III trials evaluating these agents as first-line therapy for metastatic colorectal cancer have been disappointing. Although DPD-inhibiting oral fluoropyrimidines have some activity in colorectal cancer and oral administration provides significant convenience advantages, the inferior efficacy of UFT/leucovorin and eniluracil/5-FU versus 5-FU/leucovorin in phase III trials does not support the use of these compounds. A feasible regimen for the phase III development of S-1 outside Japan has not been defined. Thus the DPD-inhibiting oral fluoropyrimidines have failed to fulfill their early promise: clinical data indicate that none of these compounds is likely to improve outcomes for patients with metastatic colorectal cancer. PMID- 14551503 TI - Second-line treatment in advanced colon cancer: are multiple phase II trials informative enough to guide clinical practice? AB - This article reviews the available data regarding the activity of second-line chemotherapy following 5-fluorouracil (5-FU), irinotecan (CPT-11) or oxaliplatin (OXA) alone or in combination. Studies undertaken in this setting, published both as full papers and in abstract form, were critically analyzed. The main conclusion is that clinical research for second and subsequent lines of treatment in advanced colon cancer (ACC) clearly needs to be optimized. A large number of small, non-randomized phase II trials have been reported without definitive conclusions. Efficient conduct of a limited number of high-quality randomized phase II trials with validation of promising regimens via phase III studies seems a preferable approach. This would not only accelerate the evaluation of new therapeutic options, but also, and more importantly, limit the number of patients receiving suboptimal treatments. The responsibility of this indispensable and urgent task lies with all researchers in this field and their partners in the pharmaceutical industry. One means to implement this approach is through strict selection of studies to be both presented and published, encouraging the spread of information provided by statistically well-designed and well-conducted trials that will eventually lead to the definition of the best standard of care for ACC patients. The conduct of repetitive phase II trials that test minor variations in dose and schedule, while commonplace, does little to advance the field. PMID- 14551504 TI - Chemosensitivity testing for gastrointestinal cancer: survival benefit potential and limitations. AB - Chemosensitivity testing is considered by some to be a useful method for predicting drug sensitivity of tumor tissues after surgery for gastrointestinal cancer. Although survival benefit is not fully established, several chemosensitivity testing methods have been used clinically, both in the selection of adjuvant therapy and in the treatment of metastatic disease. Chemosensitivity testing is used not only for determination of drug resistance, but also for determination of drug sensitivity conferring a potential survival benefit. Previous retrospective correlation studies showed survival of patients treated with a 'tested' drug to be superior to that of patients treated with a standard drug, but the clinical benefit of chemosensitivity testing in comparison to surgical therapy alone or standard chemotherapy has not been documented in a randomized controlled trial. The clinical usefulness of individualized versus standard therapy needs to be determined. Here we discuss the potential survival benefit and current limitations of chemosensitivity testing in patients with gastrointestinal cancer. PMID- 14551505 TI - The importance of prophylactic management of chemotherapy-induced neutropenia. AB - The development of colony-stimulating factors (CSFs) has provided clinicians with a valuable tool for proactive management of chemotherapy-induced neutropenia. However, clinicians are also presented with the challenge of appropriately targeting this treatment to patients at serious risk of neutropenic complications, while maintaining an economic approach to prescribing. This article discusses the seriousness of chemotherapy-induced neutropenia and reviews current approaches to the management of this condition. Febrile neutropenia risk models, new therapy options and international guidelines for the use of CSFs are also discussed. PMID- 14551506 TI - Adverse reactions to oxaliplatin: a retrospective study of 25 patients treated in one institution. AB - We reviewed the records of 25 colon cancer patients consecutively treated with an oxaliplatin-containing regimen. We differentiated between hypersensitivity reactions and pain reactions due to oxaliplatin. The patients did not receive preventive pre-medication. Four patients underwent an adverse reaction. Three patients fulfilled the criteria of a hypersensitivity reaction with tachycardia, chills and hyperhidrosis. In addition, two patients suffered from severe abdominal and chest pain. Reactions occurred during or shortly after the oxaliplatin infusion. All patients recovered under symptomatic therapy. After reacting for the first time, pre-medication was applied prior to the oxaliplatin infusion. However, due to further reactions, the treatment protocol had to be changed in all cases into a regimen not containing oxaliplatin. We conclude that adverse reactions are relatively frequent toxic side-effects of oxaliplatin, mainly in heavily pre-treated patients. Pre-medication was ineffective in preventing further reactions and consequently the treatment regimen had to be changed in all cases. PMID- 14551507 TI - Carboplatin-based chemotherapy in patients with gynecological malignancies on long-term hemodialysis. AB - We report on three cases of long-term dialysis patients with gynecological malignancies who were successfully treated with chemotherapy. Two epithelial ovarian carcinoma patients were treated with a single agent, carboplatin (100-200 mg/m2). One recurrent endometrial carcinoma patient was treated with carboplatin (200 mg/m2) and paclitaxel (135 mg/m2). Hemodialysis was started 2 h after the carboplatin infusion and lasted 4 h in all three cases. All patients tolerated these therapies without significant myelosuppression or severe side-effects. Our findings suggest these regimens are feasible, and the combination of paclitaxel and carboplatin is effective chemotherapy when administered to long-term hemodialysis patients with recurrent endometrial carcinoma. PMID- 14551508 TI - High-dose i.v. granisetron for the prevention of chemotherapy-induced emesis: cardiac safety and tolerability. AB - This phase II trial assessed the cardiovascular safety and tolerability of high dose granisetron for the treatment of nausea and vomiting in cancer patients undergoing emetogenic chemotherapy. Forty-one patients were given 30-min infusions of granisetron, 40 or 120 microg/kg i.v., as either a single dose or as split doses, at 6-h intervals. Subsequently, patients had the option of the alternative dosing regimen or to return to conventional antiemetic therapy. Patients were monitored for 24 h following the first granisetron infusion. Electrocardiogram (ECG; lead II and Holter monitoring) measurements were made during the study and blood samples for pharmacokinetic analysis were taken at regular intervals for 48 h after the start of the first granisetron infusion. During the first chemotherapy session, granisetron was administered as: (i) bolus doses of 80 microg/kg (n=3) and 120 microg/kg (n=19) or (ii) split doses of 2x40 microg/kg (n=1) and 3x40 microg/kg (n=18). Crossover therapy was administered to 22 patients, with granisetron doses of 120 microg/kg (n=12), 2x40 microg/kg (n=1) and 3x40 microg/kg (n=9). We conclude that supra-therapeutic doses up to 120 microg/kg granisetron had no clinically significant effect on ECG, pulse rate or blood pressure. The treatment was well tolerated with no significant changes in biochemistry or hematological parameters. PMID- 14551509 TI - Weekly high-dose 5-fluorouracil as 24-h infusion and folinic acid (AIO) plus irinotecan as second- and third-line treatment in patients with colorectal cancer pre-treated with AIO plus oxaliplatin. AB - Our objective was to evaluate the efficacy and safety of high-dose 5-fluorouracil (5-FU) as a 24-h infusion and folinic acid (FA) (AIO regimen) plus irinotecan (CPT-11) after pre-treatment with AIO plus oxaliplatin (L-OHP) in colorectal carcinoma (CRC). Twenty-six patients with non-resectable distant CRC metastases were analyzed for second- or third-line treatment with AIO plus CPT-11 after pre treatment with AIO plus L-OHP. On an outpatient basis, the patients received a treatment regimen comprising weekly 80 mg/m2 CPT-11 in the form of a 1-h i.v. infusion and 500 mg/m2 FA as a 1- to 2-h i.v. infusion, followed by 2000 mg/m2 5 FU i.v. administered as a 24-h infusion once weekly. A single treatment cycle comprised six weekly infusions followed by 2 weeks of rest. A total of 26 patients received 344 chemotherapy applications with AIO plus CPT-11. The main symptom of toxicity was diarrhea (NCI-CTC toxicity grade 3+4) occurring in five patients (19%; 95% CI 7-39%). Nausea and vomiting presented in two patients (8%; 95% CI 1-25%). The response rate of 26 patients can be summarized as follows: partial remission: n=7 (27%; 95% CI 12-48%); stable disease: n=9 (35%; 95% CI 17 56%) and progressive disease: n=10 (38%; 95% CI 20-59%). The median progression free survival (n=26) was 5.8 months (range 3-13), the median survival time counted from the treatment start with the AIO plus CPT-11 regimen was 10 months (range 2-24) and counted from the start of first-line treatment (n=26) was 23 months (range 10-66). We conclude that the AIO regimen plus CPT-11 is practicable in an outpatient setting and well tolerated by the patients. Tumor control was achieved in 62% of the patients. The median survival time was 10 months and the median survival time from the start of first-line treatment (n=26) was 23 months. PMID- 14551510 TI - Imatinib mesylate (STI571; Glivec)--a new approach in the treatment of biliary tract cancer? AB - Non-resectable biliary tract cancer is associated with poor prognosis due to widespread resistance to chemotherapeutic agents and radiotherapy. It is therefore essential to explore new therapeutic approaches like the inhibition of tyrosine kinases. The aim of this study was to determine the expression of c-kit and platelet-derived growth factor (PDGF) receptors (PDGFRs) and the effects of the tyrosine kinase inhibitor imatinib +/- 5-fluorouracil (5-FU) on proliferation and apoptosis in biliary tract cancer cell lines. The expression of c-kit and PDGFR mRNA was examined in 12 biliary tract cancer cell lines using RT-PCR. Cells were treated with imatinib (1, 10, 20 and 50 micromol/l) +/- 5-FU (0.1 microg/ml) for 6 days and inhibition of cell growth was assessed by manual cell counting. Cell proliferation and apoptosis were analyzed by flow cytometry of BrdU and Annexin-V/propidium iodide-stained cells. c-kit and PDGF mRNA expression was detected in 50 and 75%, respectively. Imatinib (10 and 20 micromol/l) alone inhibited cell growth significantly higher in c-kit+ cell lines (p<0.02) and inhibition was independent of PDGFR status. The combination with 5-FU increased the effect of imatinib mesylate in all cell lines. Treatment of cells with imatinib +/- 5-FU was associated with a significant induction of apoptosis, but no inhibition of proliferation. We conclude that imatinib alone exerts marked effects on c-kit+ biliary tract cancer cell lines only at intermediate and high concentrations, but there is a potential role of low-dose imatinib in combination with 5-FU for the treatment of biliary tract cancers. PMID- 14551511 TI - Induction of colon cancer cell death by 7-hydroxystaurosporine (UCN-01) is associated with increased p38 MAPK and decreased Bcl-xL. AB - UCN-01, a selective inhibitor of protein kinase C, is known to inhibit the growth of cancer cells. Although it is currently undergoing clinical evaluation, information about its effect on human colon cancer is limited and the mechanism responsible is lacking. The objective of this study was to examine the cytotoxicity of UCN-01 to human colon cancer cells in vitro and its effect on the apoptotic molecules. HT-29, a radiation- and chemotherapy-resistant human colon cancer cell, was used in the study. Cell death/apoptosis was determined by the MTT assay and DNA fragmentation measurement. NF-kappaB activity was measured by an enzyme immunoassay method. Western blot was employed to examine the expression of relevant apoptotic molecules. The result showed that UCN-01 could induce apoptosis of human colon cancer cells in a time- and dose-dependent manner. It markedly reduced the expression of Bcl-xL, but enhanced the level of p38 MAPK. In addition to Bcl-xL and p38 MAPK, UCN-01 also increased both caspase-3 and peroxisome proliferator activated receptor gamma protein levels. HT-29 cells transfected with exogenous Bcl-xL showed a significant increase in NF-kappaB activity and prevented apoptosis induced by UCN-01. The overexpression of Bcl-xL also reversed other relevant molecular changes observed in UCN-01-treated cells. In conclusion, UCN-01 exerted an antitumor effect in human colon cancer cells by inducing apoptosis. The mechanism responsible appeared to be related to reduction of Bcl-xL and increased p38 MAPK. The overexpression of Bcl-xL can significantly prevent apoptosis induced by UCN-01. PMID- 14551513 TI - The rational phase of therapeutic angiogenesis. AB - Largely disappointing results from early clinical trials have settled down the initial unrealistic hope that fueled therapeutic angiogenesis. Now that this research has reached a rational phase, every concept in the theory of neo vascularization needs to be re-evaluated. Neo-vascularization in adult tissues has been described as the result of either arteriogenesis, angiogenesis or vasculogenesis. The contribution of these mechanisms to neo-vascularization in vivo can likely not be separated experimentally. All currently known growth factors are pleiotropic and induce the secondary release of other growth factors. A complete analysis of the efficacy of growth factors therefore should include parameters of arteriogenesis, angiogenesis and vasculogenesis. Current clinical studies have likely suffered from drug regimens with short exposure of a single growth factor. Although combinations of growth factors may be theoretically appealing to create robust sustained neo-vessels, preclinical and clinical study designs may become too complex. Instead, there is some evidence that prolonged exposure to a single growth factor might result in vessels that are resistant to regression. With the vast and rapidly growing body of data that is being obtained on growth factors and pro-angiogenic strategies, approaches will emerge that are more effective than the ones presently tested in the clinic. It remains imperative however, that these approaches are rationally based on fundamental and preclinical data. PMID- 14551512 TI - The bisphosphonate pamidronate is a potent inhibitor of Ewing's sarcoma cell growth in vitro. AB - The MTT assay was used to measure the effects of pamidronate, clodronate and mevastatin on the cell viability of Ewing's sarcoma cell lines 6647, CADO-ES-1, ES-2, ES-3, RD-ES, SK-ES-1, STA-ET-2.1 and VH-64. Treatment of these cells with pamidronate inhibited cell viability in a time- and dose-dependent manner. After a 72-h incubation period with 50 microM pamidronate, cell numbers were reduced by up to 80%, whereas the monophosphonate analog 3-aminopropyl phosphonate had no effect at concentrations up to 2 mM. Clodronate reduced cell viability by maximally 40% at 1 mM. These data provide the first evidence for a direct growth inhibitory effect of pamidronate on Ewing's sarcoma cells. Hence, pamidronate definitely merits a more thorough exploration into its potential use in the therapy of patients with Ewing's sarcoma. PMID- 14551514 TI - The management of unstable angina and non-ST-segment elevation myocardial infartion. AB - Patients presenting with unstable angina and non-ST elevation myocardial infarction (UA/NSTEM) have a highly variable course. Optimal management is critical because of the high risk of death or myocardial infarction (MI) in the ensuing 30 days. In this article, we review the therapeutic options available to clinicians. Anti-ischemic therapy with beta-blockers and nitrates should be considered in all patients without contraindications. Aspirin remains a cornerstone of antiplatelet therapy and has been shown to substantially reduce the risk of death or MI. Although the data are less robust, unfractionated heparin (UFH) also appears to be efficacious, and the low-molecular-weight heparin (LMWH) enoxaparin appears to be superior to UFH. The GP IIb/IIIa inhibitors, highly beneficial in the setting of percutaneous coronary intervention (PCI), should be considered in patients with continuing ischemia or other high-risk features. The ADP receptor blocker clopidogrel has been shown to be beneficial in patients who are managed conservatively and in those who undergo PCI. Lastly, a strategy of early angiography should be considered in patients with recurrent ischemia or in those who present with high-risk features such as elevated troponins or ST deviation. Thus, early risk stratification using clinical features, electrocardiographic data, and biomarkers allows identification of subgroups of patients who are not only at high risk but also enjoy the greatest benefits from these aggressive therapies and thereby enables clinicians to target these interventions most effectively. PMID- 14551515 TI - Interpretation of new treatment guidelines for non-ST-segment elevation acute coronary syndromes: "ischemia-guided" versus "early invasive" strategies. AB - The American College of Cardiology/American Heart Association Task Force on Practice Guidelines has recently published recommendations regarding the diagnosis and management of patients with non-ST-segment elevation (NSTE) acute coronary syndromes (ACS). Conventional therapy for non-ST-segment elevation acute coronary syndrome (NSTE ACS) has traditionally employed an "ischemia-guided" approach in which diagnostic cardiac catheterization and revascularization are only used in patients with objective-evidence of residual myocardial ischemia as identified by recurrent symptoms or provocative stress testing. More recent studies, however, have demonstrated improved clinical outcomes with the use of an "early invasive" approach, employing routine coronary angiography early in the patient's hospital course, followed by percutaneous coronary intervention (PCI) or coronary artery bypass graft (CABG) surgery where appropriate. Improved clinical outcomes associated with an "early invasive" strategy may have evolved as a consequence of recent advances in both adjunctive pharmacotherapy and revascularization technique. For example, use of GP IIb/IIIa inhibitors and/or low-molecular-weight heparin prior to catheterization have been shown to reduce clinical events in NSTE ACS patients, and may reduce the risk of an invasive approach by plaque passivation prior to interventional therapy. Perhaps more importantly, the combined use of GP IIb/IIIa inhibitors and intracoronary stenting may reduce the potential early hazard of an invasive approach by specifically decreasing the incidence of death and non-fatal myocardial infarction associated with percutaneous intervention. In spite of the benefits of this synergistic combination of pharmacology and mechanical revascularization, risk stratification remains important in identifying high-risk individuals most likely to benefit from an "early invasive" approach. PMID- 14551516 TI - Enoxaparin for the treatment of acute myocardial infarction with persistent ST segment elevation. AB - Enoxaparin (E) is a low-molecular-weight heparin which has been proven more effective than unfractionated heparin (UFH) for the treatment of non-ST-segment elevation acute coronary syndromes. Limited and inconclusive on the other hand, are the data on the use of E in acute myocardial infarction with persistent ST segment elevation (STEAMI). Therefore, we performed a review of the literature in order to evaluate the level of evidence relative to the efficacy and safety of E in such a clinical setting. The effect of E in STEAMI has been evaluated in 7 clinical studies, including a total of about 9500 patients. Compared to placebo, E resulted more effective on the incidence of the combined end-point of death, re infarction and recurrent angina in the study by Glick et al. and on the patency of the infarct-related artery in the AMI-SK study. Compared to UFH, E resulted more effective on the incidence of the combined end-point of death, reinfarction and unstable angina in the study by Baird et al. and of in-hospital re-infarction and refractory ischemia rates in both ASSENT-3 and ASSENT-3 PLUS, while the effect on the patency of the infarct-related artery, which was evaluated in HART II and ENTIRE-TIMI 23, resulted non univocal. Overall, bleeding complications were more frequent than with placebo and comparable to UFH, with the exception of ASSENT-3 PLUS where pre-hospital administration of E was associated with a doubled incidence of intracranial bleeding (although only in patients older than 75 years). In conclusion, the administration of E, in association with aspirin and thrombolytics, already appears a possible therapeutic option for the treatment of STEAMI, due to its good efficacy and safety profile, along with its easiness of use. However, prior to have its use recommended, the current B level of evidence of a superior efficacy and safety compared to UFH needs to be reinforced. Further-more, some open issues relative to the use of E in particular settings (aged patients, in association with glycoprotein IIb/IIIa inhibitors and during percutaneous coronary revascularization) need to be clarified. PMID- 14551518 TI - Evolving standard in the treatment of coronary artery disease. Drug-eluting stents. AB - Coronary stent implantation is the predominant method of percutaneous coronary interventions (PCI). This is to be attributed to the ease of use beside the better short and long term clinical outcome as compared to balloon angioplasty. Nevertheless, improvements in operator skill and stent technology together with better use of adjunctive pharmacological therapy have contributed to the improvement in clinical outcome. However, the main limitation of coronary stenting is still represented by in-stent restenosis (ISR) with an estimated rate of 17-32%. Thus, compared to coronary bypass surgery, the major adverse cardiac events following stent implantation are still higher and mainly represented by the need for re-intervention. The advent of drug eluting stents (DES) has led the experts to predict that with DES there will be little or no difference between PCI and coronary bypass surgery in terms of long-term outcome leading to a further expansion of indications. The clinical trial programs of the 2 available DES for clinical use (sirolimus-eluting stent, SES - Cypher and paclitaxol eluting stent - Taxus) have been able to demonstrate the safety and clinical efficacy of both. Nevertheless, off-label use in patients on high risk for restenosis confirmed these data. At least for SES as was demonstrated by 2 "real world" registries. Thus, the introduction of DES represents a remarkable evolution for new standards in coronary artery disease treatment and offers hope to those patients considered to be "high risk" such as diabetics, patients with ISR, diffuse disease in whom surgery was previously the only therapeutic option. This paper will discuss the main results of the clinical trial programs of the DES (mentioned above) available for clinical use in the present time and analyze technical and procedural aspects which could affect long term outcome. PMID- 14551517 TI - Sirolimus-eluting stents for the treatment of in-stent restenosis. AB - The treatment of in-stent restenosis (ISR) remains one of the major therapeutic challenge for the interventional cardiologist. All percutaneous mechanical approaches have shown disappointing results and the recurrence of ISR was reported to be unacceptably high. Currently, the only proven effective therapy available for the treatment of ISR, at least for the most complex lesions, is vascular brachytherapy. However, this therapy is limited by potential side effects and logistic requirements. The introduction of drug-eluting stents, that carry and release antiproliferative agents, have demonstrated to virtually eliminate ISR in de novo lesions. In the light of this promising results for de novo lesions, sirolimus-eluting stents (SES) were recently used for the treatment of ISR in 2 pilot studies. In Sao Paulo, 25 patients with ISR treated with SES (1.4 stent per lesion) presented 4% ISR and no clinical events at 1 year. In Rotterdam, 16 patients with severe ISR were treated with 26 SES. Intravascular ultrasound evaluation demonstrated successful inhibition of neointimal hyperplasia with 1.1% volume obstruction of the stent, which is similar to the Sao Paulo series (0.8%). At 9 months clinical follow-up, 3 patients had experienced 4 major adverse cardiac events (2 deaths and 1 acute myocardial infarction necessitating repeat target vessel angioplasty). With the results presently available, SES implantation can be considered safe and potentially efficacious in the treatment of ISR. However, multicenter, long-term randomized studies are warranted in order to evaluate this new treatment concept. PMID- 14551519 TI - Using intravascular ultrasound with drug-eluting stents. What have we learned so far? AB - Drug-eluting stents (DES) promise to change the landscape of interventional cardiology, overcoming restenosis that is the major limitation of percutaneous coronary interventions. Intravascular ultrasound (IVUS) examination has been at the centre of our efforts to understand the mechanisms and define different treatment strategies during coronary interventions. IVUS interrogation and 3 dimensional IVUS measurements have been used to better define the mechanisms of benefit and potential drawbacks of DES. The findings of these studies are summarized in this article and the potential importance of IVUS in the era of DES is discussed. Evidence of neointimal hyperplasia (IH) suppression and assessment of any edge effect or vessel remodeling after implantation of DES has been evaluated by IVUS. The overall clinical importance of IVUS in the new era will depend on the amount and the clinical significance of any unsolved questions we will face and on its ability to provide answers to the evolving questions. PMID- 14551520 TI - Significance of elevated cardiac troponin after percutaneous coronary interventions. AB - Assays for cardiac troponin have become a standard for the diagnosis of myocardial damage. Due to their high sensitivity, minor myocardial injury can be frequently detected following percutaneous coronary interventions (PCI). Minor elevations in cardiac enzymes after apparently successful PCI are rather common and even modest increases in creatine kinase myocardial band (CK-MB) elevation identify a population with worse long-term prognosis compared to patients with no enzyme elevation. The significance of troponin elevation in acute coronary syndromes and its prognostic implications on short- and long-term clinical outcomes were previously demonstrated in several clinical studies. Conversely, data regarding troponin elevation after an apparently successful PCI is limited and their relevance in terms of predicting clinical outcomes remains unclear. Given the higher sensitivity of troponin essays, the incidence of troponin elevation after PCI is higher than that of CK-MB. In this review we discuss the significance and implications of post-procedural troponin elevation. PMID- 14551521 TI - Acetylcysteine for the prevention of radiocontrast -induced nephropathy. AB - Radiocontrast-induced nephropathy is the 3rd most common cause of in-hospital acute renal failure after hypotension and surgery. Radio-graphic contrast media are used at a progressive rate for several diagnostic and therapeutic applications. Prevention of radiocontrast-induced nephropathy will become more important, including risk of patient impairment and costs. Radiocontrast-induced ne-phropathy is due to vasoconstriction-mediated renal medullary ischemia and direct toxic damage to renal tubular epithelial cells. These effects may in part be mediated by generation of reactive oxygen species. Several prospective, randomized, placebo-controlled studies in patients with moderate renal insufficiency showed that the prophylactic oral administration of acetylcysteine at a dose of 600 mg twice daily along with hydration prevents the reduction in renal function after radiocontrast administration. Recently, intravenous administration of acetyl-cysteine has also been shown to be effective. Use of acetylcysteine together with hydration is the treatment of choice to prevent radiocontrast-induced nephropathy. PMID- 14551522 TI - The expanding role of antiplatelet agents in coronary artery disease. A current review of aspirin, glycoprotein IIb/IIIa inhibitors, and the thienopyridines. AB - The platelet has assumed an increasingly important role in cardiovascular medicine as our understanding of the pathophysiology of acute coronary syndromes (ACS) has evolved. Plaque rupture, platelet aggregation, and thrombus formation occur as a result of complex interaction between the platelet, the endothelium, and various inflammatory cells and circulating proteins. Aspirin continues to form the foundation of any anti-ischemic regimen, but cardiologists have long recognized the need for newer, more potent antiplatelet agents. Glycoprotein IIb/IIIa receptor antagonists and thienopryidines have been developed over the past decade and now serve as powerful complements to aspirin in the prevention and treatment of coronary events. The paper will begin with a review of aspirin as well as a discussion of the concept of aspirin resistance. The rapidly expanding body of knowledge supporting the use of glycoprotein IIb/IIIa receptor blockers and thienopyridines will then be addressed, with an emphasis on reconciling recent controversies in the literature. Future advances in the treatment of coronary artery disease will likely occur as we further refine the role of these established antiplatelet drugs and develop agents that bind to novel targets in the thrombotic cascade. PMID- 14551523 TI - Platelet glycoprotein IIb/IIIa inhibitor therapy in non-ST segment elevation acute coronary syndromes. AB - Platelet glycoprotein (GP) IIb/IIIa inhibitors prevent fibrinogen binding and platelet aggregation. They decrease ischemic complications associated with non-ST segment elevation acute coronary syndromes and percutaneous coronary intervention. Meta-analyses of 6 randomized trials of parenteral GP IIb/IIIa inhibitors in patients with acute coronary syndromes suggest a significant reduction in death and myocardial infarction in high risk patients. These include patients undergoing early percutaneous coronary intervention or those with high TIMI risk score, elevated troponin values, or diabetes mellitus. Despite guideline recommendations supporting therapy for these indications, only a minority of appropriate candidates are being treated. The risk of major bleeding is small; thrombocytopenia can result from abciximab therapy. Optimal dosing strategies continue to evolve. PMID- 14551524 TI - [Omega-3 polyunsaturated fatty acids and cardiovascular diseases]. AB - Population studies indicate omega-3 that a high fish intake, such as reported in the early Eskimo and Japanese studies, is associated with a low mortality from coronary heart disease (CHD). These effects have been associated with measurable effects on a series of established and possible risk factors. In most of the later population studies much lower daily intake of fish has been observed, such intake having only small or not even measurable effects on established risk factors. Still, their association with reduced CHD mortality, particularly sudden cardiac death seem to be reasonably well established. Beneficial effects are found at a level of consumption of about 30 g per day or 1 fish meal per week compared with populations rarely or never consuming fish. Such findings are also reflected in studies including analysis of fatty acid composition in adipose tissue and cell membranes. Fish consumption may be a marker for a healthier lifestyle or, alternatively, fish consumers may be at higher self-perceived risk for CHD and are therefore eating fish to reduce their high baseline risk. Despite all these limitations, the population studies seem to indicate that a high fish intake is associated with a low mortality from CHD. GISSI-Prevenzione was conceived as a population, pragmatic trial on patients with recent myocardial infarction and it was conducted in the framework of the Italian public health system. In GISSI-Prevenzione, 11,323 patients were enrolled in a clinical trial aimed at testing the effectiveness of omega-3 PUFA and vitamin E. Patients were invited to follow Mediterranean dietary habits, and were treated with up-to-date preventive pharmacological interventions. Long-term omega-3 PUFA 1 g daily, but not vitamin E 300 mg daily, was beneficial for death and for combined death, non fatal myocardial infarction, and stroke. All the benefit, however, was attributable to the decrease in risk for overall, cardiovascular, cardiac, coronary, and sudden death. In GISSI-Prevenzione, long-term administration of omega-3 PUFA (1 g daily) significantly decreased the risk of overall (-20%), cardiovascular (-30%), and sudden death (-45%). At variance from the orientation of a scientific scenario largely dominated by the "cholesterol-heart hypothesis", GISSI-Prevenzione results indicate omega-3 PUFA (virtually devoid of any cholesterol-lowering effect) as a relevant pharmacological treatment for secondary prevention after myocardial infarction. PMID- 14551525 TI - Early statin therapy in acute coronary syndromes. AB - Patients who survive an acute coronary syndrome are at much higher risk of a recurrent event within the following year than patients with stable coronary syndromes. Risk factor modification, including statin therapy, lowers the risk of recurrent events over many years, but also to reduces the high risk of an another event within the weeks to months following the initial acute coronary syndrome. The mechanisms that contribute to this benefit are likely related to improvements in endothelial function, a decrease in vascular inflammation, and reduced prothrombotic factors. The effects of statins may be mediated by cholesterol reduction, cholesterol-independent effects (particularly by decreasing isoprenoids), and mechanisms that are independent of inhibiting HMG CoA reductase. Observational studies show an early reduction in mortality with statin therapy started before discharge from hospital after an acute coronary syndrome. Several randomized controlled trials also support a rapid reduction in the risk of recurrent events after starting statins during the hospital admission for an acute coronary syndrome. Early statin therapy is also related to improved compliance and use of statins several years after a coronary event. Thus early statin therapy may improve both early and long-term secondary prevention efforts. PMID- 14551526 TI - Balancing benefit against risk in the choice of therapy for coronary artery disease. Lesson from prospective, randomized, clinical trials of percutaneous coronary intervention and coronary artery bypass graft surgery. AB - The ageing world population faces a coming pandemic of high-risk coronary artery disease (CAD). Patients with CAD have 3 therapeutic options, which are based on objective clinical outcome: medical therapy and risk factor modification (Medicine), and 2 forms of revascularization, coronary artery bypass graft surgery (CABG), and percutaneous coronary intervention (PCI). More than 50 large (>100 patients), multicenter, prospective, randomized clinical trials (RCT) have compared these treatment options in terms of clinical benefits and patient risks. The randomized trials which demonstrated hard outcome (survival, myocardial infarction, stroke) benefits from statins, angiotensin-converting enzyme inhibition and thienopyridines have all been completed subsequent to the publication of most Medicine versus revascularization trials. These medical therapies, plus aspirin, beta-blockers, and risk factor modification, should be made available to patients regardless of the decision to revascularize, or the decision by what means (CABG or PCI). This review integrates the information from these trials, comparing the clinical benefits against the risks inherent in the 3 therapeutic options. The results of our review show that: trials of medicine versus revascularization (either CABG or PCI) support the revascularization paradox, in that the patients at highest risk of adverse outcome, from myocardial ischemia, have a hard outcome benefit (survival, MI, or stroke) from revascularization. This paradox, first seen in the Medicine versus CABG trials of the 1970s, is evident in the trials comparing fibrinolysis and other medicines, with primary PCI for ST-elevation myocardial infarction (MI). The paradox is evident in the conservative versus invasive strategy trials of non-ST-elevation MI and unstable angina, where the benefit of revascularization occurs only in high-risk subsets. The paradox often results in sicker patients, who have more to gain from revascularization, being denied it because of the elevated perception of risk (comparable to a reperfusion paradox in ST-elevation MI, where patients most likely to benefit from thrombolytics are denied them because of the perception of risk). Trials that compared medicine with revascularization for the treatment of acute MI support the use of PCI as the preferred early stabilization strategy (90% of all PAMI trial patients). The majority of the PCI versus CABG trials enrolled populations that were at relatively low risk for ischemic clinical events. These trials demonstrated few hard outcome (survival, MI, or stroke) differences between CABG and PCI. On the basis of the results obtained the following conclusions may be drawn: medicines are the primary options for stable, low-risk CAD, and should be given to all CAD patients. Medically refractory is a useful high-risk marker of potential benefit from revascularization. CABG continues to be the complete revascularization option for patients with multivessel, multi-lesion CAD, in part because of its application to chronic occlusions. PCI is the acute stabilization method of choice for patients with on-going ischemia and acute MI, especially among patients with hemodynamic compromise, and/or major comorbidity. PMID- 14551527 TI - Coronary artery bypass surgery and percutaneous transluminal coronary angioplasty in patients with multivessel disease. AB - Since its introduction in the early 1970s, coronary artery bypass grafting (CABG) surgery has been proven to relief anginal symptoms in patients with severe myocardial ischemia. Percutaneous coronary intervention (PCI) was introduced in clinical practice at the end of the 1970s and this treatment has also been established to be effective. PCI has now surpassed CABG as the most common means for treating coronary artery disease (CAD). However, restenosis remains its Achille's heel. Until the mid-1990s, a coronary reintervention was needed in 35% to 40% of the patients. Since then, interventional techniques and technology have evolved with improved success in more complex and anatomical settings, and restenosis has been now consistently decreased to around 20%. Despite all these improvements, the incidence of restenosis, especially in the 1(st) year, is still an important limitation to PCI. The major determinants of restenosis are elastic recoil, negative vessel remodeling and neointimal proliferation as a response to vessel injury induced by angioplasty devices. The use of conventional stents has provided an efficient method to face the first 2 problems, but neointimal proliferation is not affected by stenting. A new approach consists of using the stent as a drug carrier to the target site in order to inhibit restenosis. The first results of utilization of these fascinating drug-eluting stents (DES) to treat relatively simple lesions are very promising, but further analyses for more complex lesions such as those commonly found in daily practice are needed before any definitive conclusions can be made. PMID- 14551528 TI - Normal vascular development in mice deficient in endothelial NO synthase: possible role of neuronal NO synthase. AB - PURPOSE: Nitric oxide formation by nitric oxide synthase (NOS) has been implicated in vascular injury and retinal neovascularization during oxygen induced retinopathy. However, the role of NOS in normal retinal vascular development and growth has not been studied. The purpose of these experiments was to characterize the expression of NOS in relation to vascular development and to determine the effect of deleting endothelial NOS (eNOS) on this process. METHODS: Retinal vascular development was analyzed in 150 eNOS+/+ and eNOS-/- mice ranging from 1 day to 6 months old by using a combination of morphometric and biochemical approaches. The pattern of vascular development was analyzed in retinal tissue sections and whole-mount preparations labeled with fluorescein-conjugated Griffonia simplicifolia lectin. Analysis of vascular density and arterial diameter were performed with the lectin-labeled whole-mounts using computer assisted morphometry. NO production was quantified by measuring retinal levels of nitrate/nitrite accumulation using the Greiss reaction. Western blotting techniques with isoform-specific NOS antibodies were used to evaluate differences in levels of NOS protein expression. Retinal distribution of nNOS was characterized using nNOS immunocytochemistry and NADPH diaphorase histochemistry. RESULTS: These analyses showed that the rate and pattern of retinal vascular development in eNOS-/- mice were comparable with those in wild-type control mice. Measurement of vascular density showed no significant differences between the two strains. The amount of NO production in the eNOS-/- retina was also equivalent to that in the eNOS+/+ retina. Analysis of nNOS expression within the eNOS+/+ and eNOS-/- mice showed similar levels of total nNOS protein in the two strains. Inducible NOS was not detected in either strain. Studies of nNOS distribution showed intense labeling of the deep capillary plexus in the eNOS-/- retina. This was not seen in the wild-type retinas. The number of neuronal cells showing NADPH diaphorase activity was also significantly increased in the eNOS-/- mice. CONCLUSIONS: Development of the retinal vasculature occurs normally without eNOS. The observations of similar levels of NO production, perivascular redistribution of nNOS and increased numbers of NADPH-diaphorase reactive neurons in the eNOS-/- retinas suggest that increases in vascular-associated nNOS activity compensate for the eNOS deficiency in the developing mutant retina. PMID- 14551529 TI - Gene expression profiling in human age-related nuclear cataract. AB - PURPOSE: To identify genes that are differentially expressed in age-related nuclear cataracts compared to transparent human lenses. METHODS: Total RNA was extracted from pools of central 5 mm capsulorrhexis epithelial samples microdissected at surgery from eyes with nuclear cataract or from age-matched transparent lenses (post-mortem). mRNA levels in the two samples were compared by hybridization to DNA microarrays (GeneFilter GF211) containing 4,132 known human genes. Only mRNAs consistently modulated over four comparisons were retained for analysis. A subset of the mRNA expression differences thus identified were verified and confirmed by Real-Time RT-PCR. Expressed and modulated genes were categorized according to the Gene Ontology classification. RESULTS: The data revealed 262 genes that are downregulated and 7 that are upregulated by a factor of 2.5 or more in epithelial samples from cataractous lenses compared with transparent lenses. The highest content of downregulated genes was found in the functional classes "Signal transduction", "Regulation of cell proliferation", and "Protein modification". The "Response to oxidative stress" class was one of the least modulated. Among downregulated genes, we found several mRNAs coding for transcription/translation-related proteins, heat shock proteins 70 and 27, two ubiquitin-conjugating enzymes, two subunits of the cytoskeletal/chaperone protein, tubulin, betaA4-crystallin, and a group of Alzheimer-related proteins, including presenilin 1 and presenilin 2. CONCLUSIONS: An extensive mRNA downregulation accompanies the development of the nuclear type of human age related cataract. A few genes and classes of genes more prominently displaying this type of response have been identified. Altogether, the data indicate the tendency, in age-related nuclear cataract, towards a shutdown of de novo RNA and protein biosynthesis rather than an upregulation of cell defense components such as chaperones and various kinds of antioxidative or detoxifying proteins. PMID- 14551531 TI - Identification of genes expressed in a human scleral cDNA library. AB - PURPOSE: Clones established from a human scleral cDNA library were systematically sequenced. Public database sequence comparisons were performed to generate a profile of genes expressed in the human sclera and identify candidate genes for inherited diseases with scleral involvement. METHODS: A directionally cloned pCMV PCR cDNA library was constructed from RNA isolated from scleras of human donor eyes with known plano refractive history. Plasmid DNA was extracted from randomly selected cDNA clones, and the insert sequences were determined by 5' end single pass sequencing. Expressed sequence tags (ESTs) were generated and analyzed with the GenBank BLASTN program to identify sequence homologies to known genes. RESULTS: A total of 609 ESTs underwent BLAST analysis. Of these, 341 (56%) matched 228 known human genes and 4 non-human genes, 252 matched uncharacterized ESTs, and 16 showed no significant homology to human or non-human known sequences. The most redundant connective tissue-related genes were alphaA crystalline, Xalpha-1 collagen, and beta-5 integrin. Other extracellular matrix gene matches were biglycan, syndecan, decorin, fibromodulin, proline arginine rich end leucine-rich repeat protein, transgelin, TIMP-1, and fibulin 1. Human scleral expression of all but decorin and biglycan has not previously been reported. CONCLUSIONS: This effort provides the first partial list of genes expressed in human sclera. Identification of genes expressed in the sclera contributes to our understanding of scleral biology, and potentially provides positional candidate genes for scleral disorders such as high myopia. PMID- 14551530 TI - Identification and functional clustering of global gene expression differences between human age-related cataract and clear lenses. AB - PURPOSE: Age-related cataract is a multi-factorial disease with a poorly understood etiology. Numerous studies provide evidence that the human eye lens has evolved specific regulatory and protective systems to ameliorate lens damage associated with cataract. Other studies suggest that the presence of cataract is associated with the altered expression of specific genes including metallothionein IIa, osteonectin, transglutaminase 2, betaig-h3, multiple ribosomal proteins, ADAM9, and protein phosphatase 2A. Here, we sought to identify further gene expression changes that are associated with cataract and to cluster the identified genes into specific biological pathways. METHODS: Oligonucleotide microarray hybridization was used to analyze the full complement of gene expression differences between lens epithelia isolated from human age related cataract relative to clear lenses. The expression levels of a subset of the identified genes were further evaluated by semi-quantitative RT-PCR. The identified genes were functionally clustered into specific categories and the probability of over-representation of each category was determined using the computer program EASE. RESULTS: 412 transcripts were observed to be increased and 919 transcripts were observed to be decreased by 2 fold or more in lens epithelia isolated from age-related cataract relative to clear lenses. Of these, 74 were increased and 241 were decreased at the 5 fold level or greater. Seventeen genes selected for further confirmation exhibited similar trends in expression when examined by RT-PCR using both the original and separately prepared clear and cataract RNA populations. Functional clustering of the identified genes using the EASE bioinformatics software package revealed that, among others, transcripts increased in cataract are associated with transcriptional control, chromosomal organization, ionic and cytoplasmic transport, and extracellular matrix components while transcripts decreased in cataract are associated with protein synthesis, defense against oxidative stress, heat-shock/chaperone activity, structural components of the lens, and cell cycle control. CONCLUSIONS: These data suggest that cataract is associated with multiple previously identified and novel changes in lens epithelial gene expression and they point to numerous pathways likely to play important roles in lens protection, maintenance, and age related cataract. PMID- 14551532 TI - Genetic alterations on chromosome 19, 20, 21, 22, and X detected by loss of heterozygosity analysis in retinoblastoma. AB - PURPOSE: To explore the presence of common genetic alterations in retinoblastoma and to localize the altered genomic regions. METHODS: Genetic analysis included determinations of the loss of heterozygosity (LOH) and microsatellite instability (MSI) on chromosomes 19, 20, 21, 22, and X. Investigations were carried out among 15 microdissected retinoblastoma tumors and corresponding genomic DNA specimens. RESULTS: Among the 15 retinoblastoma cases, 73% (11/15) showed genome instability (LOH and/or MSI) at one or more loci on the 5 chromosomes, although loci with recurrent LOH was infrequent. The loss of a single allele was more frequent in chromosomes 19 (33%) and 20 (27%) than the other 3 chromosomes. Five loci with recurrent allelic loss were identified, among them the most frequent allelic losses were between D19S902 and D19S571 on 19q13 and were identified in 3 out of the 15 tumor specimens. The results suggested that gene loci in the 19q13 region may be associated with tumor development in retina. In addition, 3 specimens showed moderate frequency of LOH and/or MSI in more than 6 microsatellite markers, indicating genomic instability to occur at least in a subset of retinoblastoma. CONCLUSIONS: Our results provide the first evidence of LOH in chromosomes 19 and 20 in retinoblastoma. They also support the proposition that presence of genome instability in retinoblastoma may play a role in the tumorigenesis or progression of retinoblastoma. PMID- 14551533 TI - 7-ketocholesterol stimulates differentiation of lens epithelial cells. AB - PURPOSE: To establish if oxysterols stimulate differentiation of lens epithelial cells (LEC). METHODS: Primary cultures of lens epithelial cells were incubated with 7-ketocholesterol (7-keto), 25-hydroxycholesterol (25-OH) or cholesterol at 10 microg/ml for 10 days. Cells incubated with 100 ng/ml basic fibroblast growth factor (b-FGF) were used as positive controls for differentiation. The expression of the differentiation marker p57KIP2, proliferation marker PCNA (Proliferating Cell Nuclear Antigen) and fibers specific proteins gamma-crystallin, CP49, MIP26 following treatment with oxysterols was determined by western blot. Differentiation into fiber cells was further confirmed by counting the number of lentoid bodies formed following incubation with 7-keto. RESULTS: LEC incubated with 7-keto presented higher levels of p57KIP2 and showed expression of fiber specific proteins such as MIP26 and CP49, compared to cells incubated with 25-OH or cholesterol. The differentiation marker p57KIP2 increased over time for cells incubated with 7-keto while there was a decline on the amount of the proliferation marker PCNA. The expression of the fiber specific proteins gamma crystallin, MIP26 and CP49 was detected after 5 days of incubation with 7-keto. Differentiation was accompanied by a seven-fold increase in the number of lentoid bodies formed. CONCLUSIONS: Results show for the first time that 7-keto inhibits proliferation and stimulates differentiation of lens epithelial cells into fiber cells. The presence of 7-keto in the lens may disrupt the highly regulated differentiation program of LEC, compromising normal lens growth and transparency. PMID- 14551535 TI - Effect of quiescence on integrin alpha5beta1 expression in human retinal pigment epithelium. AB - PURPOSE: The retinal pigment epithelium (RPE) is differentiated and mitotically inactive in the normal eye, but several pathologies such as proliferative vitreoretinopathy (PVR) cause RPE cells to dedifferentiate and resume proliferation. Integrins, a family of cell surface glycoproteins that mediate cell proliferation and differentiation, are thought to play fundamental roles in PVR. The aim of this study was to evaluate protein expression and gene regulation of the integrin alpha5 subunit in proliferating and quiescent RPE cells. METHODS: Protein expression was studied in situ by immunohistochemistry and in vitro at different cell confluences by immunoprecipitation. Semi-quantitative RT-PCR and transient transfections were used to determine whether increasing cell confluence also affected alpha5 subunit mRNA levels and promoter activity, respectively. RESULTS: We demonstrated that the integrin alpha5 subunit is present at the RPE cell surface both in situ and in vitro, and that alpha5 protein level is influenced by confluence. Levels of integrin alpha5 transcripts are similar for sub-confluent and confluent cells, and a small increase in the promoter activity was observed between sub-confluent and confluent cells. However, both the integrin alpha5 subunit transcript and the alpha5 promoter activity decreased when cells reached post-confluence. CONCLUSIONS: We demonstrated that cell confluence affected protein and gene expression of the integrin alpha5 subunit. Proliferating RPE cells expressed high levels of both the alpha5 protein and mRNA transcripts and showed a high promoter activity. However, when cells reached quiescence, alpha5 gene expression was substantially reduced and RPE cells expressed little alpha5 protein at their cell surface. PMID- 14551534 TI - Comparing the use of Affymetrix to spotted oligonucleotide microarrays using two retinal pigment epithelium cell lines. AB - PURPOSE: The present study was designed to compare the results obtained from two different microarray platforms: spotted cDNAs using a two-color system (Clontech, Atlas Glass Human 3.8) and the Affymetrix platform. We evaluated the internal consistency within each of the platforms, and compared the results across the two platforms. METHODS: RNA was isolated from two retinal pigment epithelial (RPE) cell lines, D407 cells and ARPE19 cells. Each microarray system requires a specific RNA isolation and target preparation procedure. To compare the results between the two platforms, the intensity values for each platform were standardized and scaled. This allowed for a direct comparison of the data between two very different microarray platforms. Real-time RT-PCR was used as an independent conformation of expression levels for selected transcripts. The protein levels for some of these genes were determined using a quantitative immunoblot method. RESULTS: First, we compared the transcriptome of the D407 cell line to itself. Within each of the platforms there was a high degree of consistency. However, when the data from the Atlas Glass Human 3.8 microarray platform was compared to that of the Affymetrix platform there was a dramatic lack of agreement. The second step was to compare the mRNA profile of the ARPE19 cell line to the D407 cell line. Again there was good agreement within each platform. When the results of the Atlas Glass Human 3.8 platform were compared to the Affymetrix platform, there was a surprising lack of agreement between the two data sets. Real-time RT-PCR was used as independent means of defining RNA levels in the two cell lines. In general, the real-time RT-PCR results were in better agreement with the Affymetrix platform (85%) than the Atlas Glass platform (33%). In addition, we also examined the levels of 11 proteins in these two cell lines using a quantitative immunoblot method. The results from this protein analysis had a higher degree of concordance with the results from Affymetrix platform. CONCLUSIONS: In both the Atlas Glass Human 3.8 system and the Affymetrix platform, there is a high degree of internal consistency. However, comparisons between the two platforms show a lack of agreement. In general, the real-time RT PCR confirmed the results on the Affymetrix system more often than those from Atlas Glass arrays. However, in both cases, conformation by an independent method proves to be of considerable value. PMID- 14551536 TI - Organ-specific gene expression in the rhesus monkey eye following intravenous non viral gene transfer. AB - PURPOSE: The transfer of exogenous genes to the entire retina and other ocular structures is possible with a vascular route of gene delivery using a non-viral gene transfer method. The present studies examine the extent to which either beta galactosidase or luciferase expression plasmids are targeted to the retina in the adult rhesus monkey following intravenous administration. In addition, these studies examine the pattern of organ expression of the transgene in the rhesus monkey depending on whether the plasmid is under the influence of a widely expressed promoter, the SV40 promoter, or an ocular-specific promoter, the opsin promoter. METHODS: The plasmid DNA with either the SV40 or opsin promoter is encapsulated in the interior of 85 nm pegylated immunoliposomes (PILs), which are targeted across the blood-retinal barrier and into ocular cells with a monoclonal antibody to the human insulin receptor. Following a single intravenous injection of the PIL carrying the transgene, the animals were sacrificed 2, 7, or 14 days later for the measurement of beta-galactosidase or luciferase gene expression in the monkey eye and peripheral organs. RESULTS: Histochemistry showed expression of the beta-galactosidase gene throughout the entire primate retina including the photoreceptor cells with either an SV40 or a bovine opsin promoter. Whereas the SV40 promoter enables gene expression in other organs of the primate (brain, liver, spleen), the opsin promoter restricted trans-gene expression to the primate eye, as there was no gene expressed in other organs. The retinal luciferase activity at 2 days after administration was 9.6+/-0.4 pg luciferase/mg protein, and at 14 days after administration was still comparable to maximal levels of luciferase gene expression in the mouse or rat. Confocal microscopy with antibodies to the insulin receptor and to beta-galactosidase demonstrated co localization in the retina, with high expression of the trans-gene and the insulin receptor in the inner segments of the photoreceptor cells. CONCLUSIONS: The PIL non-viral gene transfer technology makes possible adult transgenics in 24 h. Ectopic expression of exogenous genes in organs other than the target organ is made possible with the use of organ specific promoters, and gene expression in the primate is restricted to the eye when the trans-gene is under the influence of the opsin promoter. Plasmid-based gene expression is still in the therapeutic range for 2-3 weeks after a single intravenous administration. Exogenous genes are expressed throughout the entire primate retina following the delivery of the gene to the eye via a trans-vascular route. PMID- 14551537 TI - Polymorphisms in OPA1 are associated with normal tension glaucoma. AB - PURPOSE: [corrected] To confirm whether specific polymorphisms in intron 8 (IVS8) of the OPA1 gene are found more commonly in patients with normal tension glaucoma (NTG) compared to normal controls. METHODS: This is a cohort study of 61 patients with NTG, 49 known healthy controls and 119 individuals from the general population. The DNA sequence was determined at the +4 and +32 positions of IVS8 of the OPA1 gene. Hardy-Weinberg equilibrium was confirmed in our population by comparing the allele frequencies in two additional genes, TP53 and TYRP1. Genotypes for the NTG and control groups were compared for statistically significant differences. RESULTS: There were no differences in the OPA1 genotypes of the NTG and control groups at the +4 location, as had been suggested in a previous study, but a significant difference was observed at the +32 location of IVS8. The CC genotype was found in 28% of NTG patients compared to 13% of controls (p=0.006). The TC genotype was more prevalent in the control population (p=0.02) but this difference did not reach statistical significance when the Bonferroni adjustment was made for multiple analyses. CONCLUSIONS: We have refined the previously reported association between OPA1 sequence changes and NTG by identifying a specific CC genotype at position +32 in IVS8 of the OPA1 gene that acts as a marker for NTG. At the current time, NTG is frequently diagnosed late when loss of neurons has already caused significant and irreversible peripheral field loss. If a test could be designed to identify those people at risk of developing NTG, then careful screening might detect earlier signs of disease allowing commencement of treatment before significant field loss has occurred. PMID- 14551540 TI - Long-term results of arthroscopic resection of the distal clavicle with concomitant subacromial decompression. AB - PURPOSE: The goal of the study was to evaluate the long-term outcome of combined arthroscopic distal clavicle excision and subacromial decompression. TYPE OF STUDY: Retrospective, long-term cohort evaluation. METHODS: Twenty patients with an average follow-up of 6 years (range, 3.9 to 9 years) were reviewed. All patients had ipsilateral impingement syndrome and acromioclavicular joint disease at the time of surgery and underwent arthroscopic subacromial decompression combined with arthroscopic distal clavicle excision. All patients returned for evaluation in person, in addition to filling out a questionnaire incorporating the University of California, Los Angeles (UCLA), and Constant scoring systems. Preoperative and postoperative radiographs were available for all patients. RESULTS: Postoperatively, all patients had pain relief and were satisfied with the result. The average postoperative UCLA Shoulder score was 29.8 +/- 0.6, compared with 17.5 +/- 3.0 before surgery (P =.001). The Constant Shoulder score averaged 98.5 +/- 2.1 postoperatively, compared with 70.5 +/- 11.2 preoperatively (P =.001). There was 100% good to excellent results using both scoring systems. Individual components of the UCLA scoring system (pain, function, and power) all showed significant postoperative improvement (P =.001). Constant categories of pain, activities of daily living, range of motion, and power also improved. Follow-up radiographs showed maintenance of the resected distal clavicle in 19 patients. Five patients (25%) had radiographic evidence of calcific density distal to the resected clavicle but were asymptomatic. CONCLUSIONS: The long-term results of arthroscopic resection of the distal clavicle with concomitant subacromial decompression are uniformly good or excellent. Impingement and acromioclavicular joint disease frequently coexist and should be identified and treated concurrently. PMID- 14551541 TI - Internal impingement in the etiology of rotator cuff tendinosis revisited. AB - PURPOSE: The theory of internal impingement holds that, in overhead athletes, repeated contact between the undersurface of the rotator cuff and the posterosuperior glenoid rim leads to articular-sided partial-thickness rotator cuff tears and superior labral lesions. However, we have noted this same constellation of lesions in our general patient population. These recreational athletic patients do not routinely assume the position of extreme abduction and external rotation, and thus are unlikely to experience significant internal impingement forces. The goal of this study was to document the prevalence of superior labral lesions in patients being treated for partial-thickness undersurface rotator cuff tears. TYPE OF STUDY: Retrospective case series. METHODS: We retrospectively reviewed the records of 75 shoulders arthroscopically treated for partial-thickness articular-sided rotator cuff tears. With the exception of one professional tennis player, no patients were playing sports at a professional or major college level. No professional or collegiate throwing athletes were included. The prevalence of these lesions and their association with recreational athletics was noted. RESULTS: We found that 55 of 75 (73.3%) shoulders with articular-sided partial-thickness rotator cuff tears also had superior labral lesions. A statistically significant increased prevalence of superior labral lesions in the dominant shoulder was seen (P =.03). In addition, our patients who engaged in overhand throwing had significantly fewer superior labral lesions in the dominant shoulders than did nonthrowers (P =.017). CONCLUSIONS: The "kissing lesions" of undersurface rotator cuff tears and posterosuperior labral damage may be explained by mechanisms other than "internal impingement." PMID- 14551542 TI - Laser-assisted thermal capsulorrhaphy. AB - PURPOSE: The purpose of this study was to review the clinical results of laser assisted thermal capsulorrhaphy in patients with glenohumeral instability. TYPE OF STUDY: A case series of consecutive patients with shoulder instability treated with laser-assisted thermal capsulorrhaphy by one surgeon. METHODS: From 1994 through 1997, 60 shoulders in 59 patients (27 men, 32 women) with no previous shoulder surgery underwent laser-assisted thermal capsulorrhaphy. The direction of instability was classified as anterior in 30, posterior in 7, anteroposterior in 4, and multidirectional (MDI) in 19. Patients were evaluated on the basis of pain, recurrent instability, function, and satisfaction. RESULTS: Nine patients were lost to follow-up evaluation, and 9 patients underwent revision surgery (7 MDI, 1 anterior, I posterior). Of the remaining 42 patients, none underwent further surgery; the average follow-up time was 38.4 months (range, 24 to 66 months); and pain scores improved from 7.8 +/- 2.6 (1-10 scale) to 1.7 +/- 2.6. postoperatively. In addition, 86% of patients reported none or rare episodes of instability compared with 37% preoperatively. Painless overhead use of the shoulder improved from 12% to 73%. Scores based on the American Shoulder and Elbow Surgeons (ASES) rating system improved from 61.0 +/- 2.6 to 89.5 +/- 15.0 (P <.05). Patient satisfaction for the remaining 42 patients, on a 1 to 10 scale, averaged 8.0 +/- 2.8. CONCLUSIONS: Based on these results, we conclude that laser assisted thermal capsulorrhaphy is an effective adjunct in the treatment of anterior and posterior instability. Patients with MDI treated with this technique have high failure rates. We recommend caution when approaching MDI patients with this technique. PMID- 14551543 TI - Loose body in the wrist: diagnosis and treatment. AB - PURPOSE: The purpose of this study was to report on 10 cases of symptomatic loose bodies in the wrist joints diagnosed using arthroscopy. TYPE OF STUDY: Retrospective review. METHODS: From 1986 to 2000, we performed wrist arthroscopy for 707 patients, 10 of whom had loose bodies in the wrist joints. The clinical records were reviewed retrospectively. The patients included 8 men and 2 women, and the average age was 28 years (range, 16 to 67 years). The chief complaint was wrist pain in all patients, but locking was uncommon. Preoperative diagnosis was difficult in all but 3 cases; in those cases, an osseous component was found within the loose bodies. The remaining cases were diagnosed by wrist arthroscopy. RESULTS: The loose bodies existed in the radiocarpal joint in 5 cases, and all could be removed arthroscopically. In the other 5 cases, the loose bodies were in the distal radioulnar joint, and arthrotomy was needed to remove them. After removal of the loose bodies, the pain was relieved in all cases without any surgical complications. CONCLUSIONS: Loose bodies in the wrist joint should be included in the differential diagnosis for chronic wrist pain. Wrist arthroscopy is of value because the preoperative diagnosis is usually difficult. PMID- 14551544 TI - Description of a new endoscopic posterior cruciate ligament reconstruction and comparison with a 2-incision technique. AB - PURPOSE: We introduce our technique (a new route for the graft) of endoscopic posterior cruciate ligament (PCL) reconstruction and describe the advantages in comparison with the conventional surgical technique. TYPE OF STUDY: Nonrandomized control study. METHODS: The idea of this surgical technique is to minimize graft angulation at the inner edge of the bone tunnel. The tibial entry point of the guide pin is under the tibial lateral subcondylar flare, approximately 1 to 2 cm anterior to the posterior cortex. This creates less graft angulation on the posterior aspect of the tibia. A new drill system has been devised to allow antegrade femoral drilling starting from inside the notch. This method also allows better femoral tunnel orientation. As a substitute graft material, we use autogenous hamstring tendons, and we secure them with an EndoButton (Smith & Nephew, Andover, MA) and post screw. From 1992 to 1995, 43 2-incision PCL reconstructions using autogenous hamstring tendons were performed. From 1995 to 2001, 90 endoscopic PCL reconstructions using looped autogenous hamstring tendons and an EndoButton were performed. Cases were specified according to the inclusion criteria of this study. As a result, 51 patients were included in this study. The 2-incision group comprised 22 patients, and the endoscopic group comprised 29 patients. The clinical evaluation was performed using the International Knee Documentation Committee (IKDC) form. The quadriceps strength was measured using Biodex System II (Biodex, New York, NY). The period of time to achieve 90 degrees flexion after surgery was also compared. RESULTS: No significant differences were seen between the 2 groups tested with respect to the overall IKDC rating score. Side-to-side differences of anteroposterior total laxity (KT-1000, manual maximum) was 3.95 +/- 1.96 mm in the 2-incision group and 2.38 +/- 1.42 mm in the endoscopic group (P <.05). The average time to achieving 90 degrees of flexion after surgery was 16.6 +/- 8.6 days in the 2-incision group and 12.1 +/- 3.5 days in the endoscopic group. Achievement of range of motion in the endoscopic group was significantly shorter. The peak torque of isokinetic contraction in the endoscopic group was significantly greater than in the 2-incision group 1 year after surgery. CONCLUSIONS: Better posterior stability and quicker postoperative recovery of range of motion and muscle strength were advantages of the endoscopic technique over the 2-incision technique in PCL reconstruction. PMID- 14551545 TI - Articular cartilage degeneration after frozen meniscus and Achilles tendon allograft transplantation: experimental study in sheep. AB - PURPOSE: The purpose of this study was to analyze cartilage degeneration in knees after total medial meniscectomy, transplantation of fresh-frozen meniscus allograft, and Achilles tendon allograft. TYPE OF STUDY: Experimental study. METHODS: We have studied the articular cartilage in the medial compartment of the left knees in 32 sheep aged 5 to 6 months, with 8 animals in each group. The study was performed after meniscectomy (group I), transplantation of fresh-frozen meniscus allograft (group II), use of fresh-frozen Achilles tendon allograft (group III), and in a control group (group IV). For the histologic study, all samples were stained with Masson's trichrome and Safranine-O. Mankin's score was applied to grade the histologic damage to the articular cartilage. RESULTS: The group with the greatest number of degenerative changes was group III, followed by groups I and II. The percentage of thickness of cartilage detected by Safranine-O stain was found to be significantly different in both tibia and femur between the control group and the other 3 groups, but not among groups I, II, and III. The immunoreactivity of the articular surfaces in tibia and femur showed notable differences in all the groups. Collagen X was present in the degenerative hypertrophic chondrocytes in the damaged articular surfaces. CONCLUSIONS: Meniscal replacement with meniscal and Achilles tendon allografts provides partial protection against articular damage after a meniscectomy. PMID- 14551546 TI - A comparative study of medial versus lateral arthroscopic partial meniscectomy on stable knees: 10-year minimum follow-up. AB - PURPOSE: Our goal was to compare results of partial medial arthroscopic meniscectomy with results of partial lateral arthroscopic meniscectomy and to determine prognostic factors. TYPE OF STUDY: Retrospective comparative study with statistical analysis. METHODS: In this study, 362 medial and 109 lateral isolated arthroscopic meniscectomies are presented with a minimum follow-up time of 10 years. All knees were stable with no previous surgery or traumatic lesion. RESULTS: In this study, 95% of the patients were very satisfied or satisfied with the results of the medial meniscectomy, and 95.5% with results of the lateral meniscectomy (P =.32). According to grades 1 and 2 of the International Knee Documentation Committee (IKDC) form, 85.8% of the medial meniscectomy group were free of any symptoms, as were 79.7% of the lateral meniscectomy group (P =.11). Radiologic changes after medial and lateral meniscectomy were found in 21.5% and 37.5%, respectively (P =.11). The rates of radiologic changes in patients in whom the contralateral knee was radiologically normal were 22.3% and 39%, respectively (P =.016). The rate of repeat surgeries for osteoarthritis was less than 0.2%. CONCLUSIONS: Subjective and clinical results after medial or lateral meniscectomy are quite similar, but radiologic results are significantly worse after lateral meniscectomy. The most accurate way to determine the degeneration caused by the meniscectomy is to evaluate joint space narrowing in patients in whom the contralateral knee was radiologically normal. Otherwise, partial medial or lateral meniscectomy are well tolerated. A better prognosis can be predicted for a patient with an isolated medial meniscal tear with one or more of the following factors: age less than 35 years, a vertical tear, no cartilage damage, and an intact meniscal rim at the end of the meniscectomy. With an isolated lateral meniscal tear, a better prognosis can be predicted if the patient is young and has an intact meniscal rim at the end of the meniscectomy. PMID- 14551547 TI - The accuracy of joint line tenderness by physical examination in the diagnosis of meniscal tears. AB - PURPOSE: The purpose of this study was to document the accuracy of joint line tenderness in the diagnosis of meniscal tears. TYPE OF STUDY: Prospective cohort study. METHODS: There were 104 male recruits (age range, 18 to 20 years; mean, 19.2 years) with suspected meniscal lesions who underwent arthroscopy. A thorough history and physical examination was performed on each patient by a physician with 4 years' experience on arthroscopic knee surgery. Twenty-six (25%) patients sustained injuries while in the Turkish Army, and 78 patients (75%) sustained injuries before they came to the army. On physical examination, assessment of joint line tenderness at the lateral and medial joint lines was performed with the patient's knee flexed 90 degrees. The accuracy, sensitivity, specificity, and positive and negative predictive values of joint line tenderness for medial and lateral sides were calculated based on arthroscopic findings. RESULTS: In 104 knees, the diagnosis was correct in 71 (68%) and incorrect in 33 (32%) knees. A total of 37 medial meniscal tears and 27 lateral meniscal tears were identified at arthroscopy. Other pathology included 11 anterior cruciate ligament (ACL) tears, 5 medial femoral chondral lesions, 4 chondromalacia patellae, and 1 medial plica. No discernable abnormalities were seen in 21 knees. A preoperative diagnosis of a medial meniscal tear was made in 54 knees and 32 were confirmed at arthroscopy. A lateral meniscus tear was suspected in 27 knees and confirmed in 25. Five medial meniscus and 2 lateral meniscus tears were seen at arthroscopy. These had not been suspected with tenderness over the joint line. The accuracy of the test was lower with the presence of ACL lesions and condromalacia patella. CONCLUSIONS: I concluded that joint line tenderness as a test for lateral meniscal tears is accurate (96%), sensitive (89%), and specific (97%). However, for medial meniscal tears, rates are lower. PMID- 14551549 TI - Arthroscopic study of the knee joint in fetuses. AB - PURPOSE: The purpose of this study was to macroscopically examine the fetal knee joint via arthroscopy. We have attempted to identify and describe the specific characteristics of the fetal knee joint, how it evolves during the last few weeks of intrauterine development, and any possible variations with regard to the adult knee. TYPE OF STUDY: Observational anatomic case series. METHODS: We used 20 frozen fetuses with a gestational age of 24 to 40 +/- 2 weeks, obtained from spontaneous abortions. Examination was performed with standard arthroscopic surgical equipment, using a 2.7-mm optical lens. Whenever possible, we tried to use standard arthroscopic portals. Images were obtained for comparison with the adult knee. RESULTS: Suprapatellar and infrapatellar septa were an almost consistent finding. The suprapatellar septum always opened laterally and was intact in the developmentally younger specimens. We found 2 mediopatellar septa. The femoral attachment of the anterior cruciate ligament (ACL) differed in appearance from that of the adult in that it was more ribbon-like. The lateral meniscus had a more spread-out appearance than its adult counterpart, especially in the specimens of a younger gestational age. We were surprised at the easy accessibility of the popliteal hiatus and the clear arthroscopic vision we were able to obtain of the involved structures. CONCLUSIONS: To our knowledge, this is the first arthroscopic study to target the fetal knee. The results indicate minimal differences when compared with the adult knee, and for some structures, such as the popliteal hiatus, the anatomy seen was easier to discern than in adult knee arthroscopy. PMID- 14551548 TI - Evaluation of postoperative bupivacaine infusion for pain management after anterior cruciate ligament reconstruction. AB - PURPOSE: Postoperative pain control has received increasing attention by health care providers in the new millennium. In fact, pain was called the "sixth vital sign" by the Joint Commission on Accreditation of Healthcare Organizations (JCAHO) in 2001. The continued challenge of effective, safe analgesia in the outpatient setting has promoted the use of various devices designed to deliver local anesthetic directly to the surgical site. We endeavored to evaluate the efficacy of one such device currently in use. TYPE OF STUDY: Prospective, randomized, placebo-controlled, double-blinded study. METHODS: In this study, 49 consecutive patients were prospectively enrolled and randomly assigned to 1 of 3 groups after anterior cruciate ligament (ACL) reconstruction. Patients and investigators were blinded to group assignment. Group 1 (control group) received no catheter. Group 2 (placebo group) received an infusion catheter filled with saline. Group 3 (experimental group) received the same catheter filled with 0.25% bupivacaine solution. All patients received an ipsilateral femoral nerve block with 30 mL 0.25% bupivacaine and 20 mL 0.25% bupivacaine intra-articular injection. Patients recorded narcotic consumption and pain levels on visual analogue scales twice a day for 4 days after surgery. The catheters were removed on day 4 and physical therapy performance was recorded. The patients were then asked to continue to record pain ratings and medication consumption for an additional 4 days after catheter removal. All patients underwent bone-patellar tendon-bone ACL reconstruction by the senior author (P.D.F.). Seven patients were excluded from the study for ineffective femoral nerve block or catheter disconnection or occlusion. Narcotic consumption and the maximum, minimum, and median pain ratings were analyzed by analysis of variance. RESULTS: Median pain ratings show lower pain levels (P <.03) for both catheter groups versus the control group. No significant differences were found between the catheter groups for the median pain ratings, but lower maximum pain ratings were seen in the bupivacaine group compared with both placebo and no-catheter control subjects. Postoperative narcotic consumption was also lower in both catheter groups versus control subjects (P <.03). Physical therapy data revealed no difference in range of motion on postoperative day 4. More patients were able to perform straight leg raises during the first therapy session in both the saline placebo catheter group (70%) and bupivacaine group (72%) compared with the control group (50%). CONCLUSIONS: The data suggest some element of placebo benefit at median pain ratings but a protective effect of the bupivacaine at maximum pain levels. PMID- 14551552 TI - Research methodology. AB - Research aims to reach valid conclusions through scientific enquiry. Valid conclusions can only be reached if bias is minimized or eliminated. Bias can potentially take place in the design, implementation, or analysis of a study. Various study designs reduce bias, and this article reviews some of the more common study designs in orthopaedic sports medicine. We also discuss bias and confounding factors as they relate to these studies. PMID- 14551553 TI - A primer for statistical analysis of clinical trials. AB - Randomized controlled trials have become the cornerstone of current practice evidence-based medicine. In this article, we present concise descriptions of fundamental statistical concepts and methods frequently used in the analysis of randomized trials. These include descriptive statistics, statistical inferences, techniques for the comparison of means or proportions from two samples, correlation, and regression analysis methods. The uses of these methods are illustrated with a number of practical examples, and the pitfalls of these topics are also briefly discussed. Lastly, some frequently used statistical terms and their meanings are also provided. By the end of the article, the reader should have sufficient knowledge to appreciate the statistical aspects of most clinical trial reports. PMID- 14551554 TI - Factors to consider in determining the feasibility of randomized clinical trials. AB - This article briefly reviews some of the important feasibility issues to be considered, and often overlooked, prior to undertaking clinical research. Topics such as time requirements, the multidisciplinary approach, the importance of a Clinical Research Coordinator, training of personnel, enrollment issues, funding, and the intrinsic and extrinsic benefits of conducting clinical research are discussed. PMID- 14551555 TI - Locked knee caused by meniscal subluxation: magnetic resonance imaging and arthroscopic verification. AB - Subluxation or dislocation of an intact lateral meniscus is a controversial and rarely reported cause of knee pain and locking. We report a case of knee locking caused by lateral meniscal subluxation in the absence of a meniscal tear or true discoid meniscus, with both magnetic resonance imaging (MRI) and arthroscopic verification. A 9.5-year-old child experienced multiple episodes of locking in full flexion of the knee. After 6 months of symptoms, arthroscopy was performed and showed no meniscal tear or a discoid meniscus. The patient's knee locking recurred after arthroscopy. MRI was performed when the patient presented acutely with the knee locked. MRI showed anterior dislocation of the posterior horn of the lateral meniscus with the knee in the locked position. The MRI was immediately repeated after the author reduced (manipulated) the locked knee into extension. On the repeat MRI, the lateral meniscus had returned to a normal position. On repeat arthroscopy, the posterior horn of the lateral meniscus was hypermobile and could be displaced into the notch and did not show a frank tear. The meniscus was repaired to the capsule with sutures. At the 2-year follow-up evaluation, the patient had no complaints and no clinical signs of locking. PMID- 14551556 TI - Acute popliteal artery occlusion after arthroscopic posterior cruciate ligament reconstruction. AB - Arthroscopic posterior cruciate ligament reconstruction may carry certain risks of complications, including injury to the neurovascular structures in the popliteal region. Acute occlusion of the popliteal artery, a limb-threatening complication, was reported after total knee arthroplasty. We report a case of acute popliteal artery occlusion after arthroscopic posterior cruciate ligament reconstruction. The possible causes included underlying artherosclerosis, the use of pneumatic tourniquet, surgical manipulation, and arterial spasm. Early diagnosis and prompt surgical treatment are the keys to success in the treatment of this limb-threatening complication. Spontaneous resolution of the thrombus such as is reported in this case is exceptional. PMID- 14551557 TI - Arthroscopically assisted quadriceps double-bundle tibial inlay posterior cruciate ligament reconstruction: An analysis of techniques and a safe operative approach to the popliteal fossa. AB - The arthroscopically assisted posterior cruciate ligament tibial inlay technique, frequently used in athletic individuals and in revision cases, requires a thorough and comprehensive understanding of posterior knee anatomy. Importantly, variations in the posterior vascular anatomy may be encountered. A safe and methodical posteromedial approach in a layered fashion to achieve proper and safe tibial inlay graft placement and fixation is described. The authors advocate use of a double-bundle quadriceps tendon autograft. Graft position of the double strands, fixation, and tensioning issues are presented. PMID- 14551558 TI - Use of autograft quadriceps tendon for double-bundle posterior cruciate ligament reconstruction. AB - Many graft choices are available for the reconstruction of the posterior cruciate ligament (PCL)-deficient knee. These choices range from multiple autograft and allograft sources. Preoperative planning must take into account the viability of knee autografts and the availability of allografts. The nature of the PCL injury must also be taken into account, such as whether only the PCL is deficient or the PCL lesion is part of a complex multiple ligament-injured knee. Our institution has begun to use the central quadriceps tendon bone autograph for multiple types of PCL reconstructions. This paper discusses the surgical techniques used to harvest and secure a double-bundle central quadriceps tendon bone autograph for PCL reconstructions using both open and arthroscopic approaches. PMID- 14551559 TI - Posterior approach for arthroscopic reduction and antegrade fixation of avulsion fracture of the posterior cruciate ligament from the tibia with cannulated screw and washer. AB - Avulsion fracture of the posterior cruciate ligament (PCL) is a rare condition, and arthroscopically assisted reattachment of the surgical fixation of the fragment is not always an easy task. Only a few reports describe techniques for arthroscopic fixation of avulsion of the PCL.We report on a case treated arthroscopically with reduction and antegrade fixation of an avulsion fracture of the tibial attachment of the PCL with a cannulated screw and washer through an additional posterolateral portal. Postoperative morbidity was reduced, and rehabilitation was accelerated. Fixation with a cannulated screw and washer is technically simple and allows for stable fixation and immediate postoperative mobilization and pain-limited weight-bearing, even in cases of a comminuted fragment. The safe zone for an additional posterolateral portal and the technique for placing instruments and a guidewire to avoid neurovascular structures is defined. PMID- 14551560 TI - Arthroscopically assisted osteochondral autogenous transplantation for osteochondral lesion of the talus using a transmalleolar approach. AB - Osteochondral autogenous transplantation for osteochondral lesions of the talar dome is usually performed through a miniarthrotomy approach, and a medial malleolus osteotomy approach is selected if the lesion is located posteromedially. Recently, we used a transmalleolar approach without osteotomy of the medial malleolus to perform osteochondral autogenous transplantation for a lesion located in the posteromedial portion of the talar dome. We report on this technique and discuss its advantages, disadvantages, and indications. PMID- 14551562 TI - Is bone tunnel osseointegration in hamstring tendon autograft anterior cruciate ligament reconstruction important? AB - A 27-year-old man underwent anterior cruciate ligament (ACL) reconstruction using 4-strand hamstring autograft with femoral and tibial interference screw fixation. Four weeks after surgery, he developed a discharging hematoma through the graft harvest-tibial tunnel incision, which persisted. The patient required further surgical intervention 7 weeks after the initial surgery. The wound was debrided, the tibial interference screw was removed, and the tibial tunnel was completely cleared of graft remnants. Arthroscopy of the knee was performed, in which the ACL graft appeared healthy and viable. No evidence of intra-articular sepsis was found. Postoperatively, the rehabilitation program was uneventful and, at 36 months, the patient has unrestricted activity and no clinical evidence of excessive ACL laxity. This case supports the importance of marginal articular surface healing of the ACL graft, suggesting that tibial intratunnel healing becomes redundant. PMID- 14551563 TI - Snapping annular ligament of the elbow joint in the throwing arms of young brothers. AB - We examined young brothers with symptomatic snapping elbow in the throwing arm. Arthroscopic examination confirmed the mechanism of snapping, in which loose and protruded annular ligament-like tissue covered the volar half of the radial head in elbow extension and uncovered the radial head in deep elbow flexion. Arthroscopic resection of the annular ligament-like tissue was performed in one brother. Histologic examination of the removed tissue showed degenerated ligament tissue. Excision of loose annular ligament abolished snapping. Contralateral elbows of the brothers also showed similar asymptomatic snapping. Researchers suggest that a hereditary factor contributing to loose annular ligament and repetitive microtrauma from throwing is the cause of symptoms. PMID- 14551564 TI - Failed meniscus repair. AB - This report describes problems arising after an arthroscopic meniscal refixation using the biodegradable Clearfix screw system (Innovasive Devices, Marlborough, MA). A serious problem arose in the form of migration of a meniscal screw that had been inserted in accordance with the manufacturer's instructions. Dislocation of the screw led to pain on the medial aspect of the knee joint and irritation of the infrapatellar branch of the saphenous nerve. The case reported here is a first analysis of the potential complications associated with the use of this meniscal refixation system. PMID- 14551565 TI - The mini vent technique: a simple method to facilitate accurate secondary portal placement in shoulder arthroscopy. AB - Accurate portal placement is crucial in diagnostic and therapeutic shoulder arthroscopy. However, knowledge of anatomy and surgical principles may not be enough. Placement of a second portal is often hindered by a small amount of bleeding. Our technique easily rectifies this frequent problem by using a simple mini-vent. PMID- 14551566 TI - Ochronotic arthropathy: arthroscopic findings in the shoulder and the knee. AB - Ochronotic arthropathy is a disorder resulting from the deposition of homogentisic acid derivatives in the articular cartilage and the menisci. Large joints of the appendicular skeleton are preferentially affected. The clinical picture resembles that of degenerative joint disease. We present the arthroscopic findings in the shoulder and the knee in a 40-year-old man with ochronotic arthropathy and discuss the role of arthroscopy in the diagnosis and management of this rare metabolic disorder. PMID- 14551567 TI - A broken sewing needle in the knee of a 4-year-old child: is it really inside the knee? AB - We report on a case of a broken needle that migrated inside the knee joint of a 4 year-old girl. Searching for any small foreign body in the knee joint is not easy in either open or arthroscopic procedures. In this case, the surgery was made more difficult because of technical delays and diagnostic difficulties in defining the surgical plane of the needle. Arthroscopic expertise and some basic precautions can minimize the morbidity to a young patient and prevent migration into the knee joint proper. PMID- 14551568 TI - Bilateral patellar component dissociation in a patient with total knee arthroplasties. AB - We present a case of bilateral patellar component dissociation 6 years after bilateral total knee arthroplasty. The patient had undergone arthroscopic lateral releases bilaterally for patellar maltracking. After repetitive trauma, the patient experienced patellar component dissociation, which was treated arthroscopically by removing the patellar components and leaving the patellae unresurfaced. The patient's symptoms improved substantially. Our case highlights certain features of the etiology as well as the management of patellar component dissociation in the total knee arthroplasty, showing the important role of arthroscopy. PMID- 14551572 TI - Phosphodiesterase type 5 inhibitors: a biochemical and clinical correlation survey. AB - Phosphodiesterase type 5 (PDE 5) is the major cGMP hydrolyzing enzyme in penile corpus cavernosum and is an important regulator of nitric oxide-mediated smooth muscle relaxation. The critical role of PDE 5 in penile erection and the recent availability of specific and potent inhibitors of PDE 5 have enabled the development of effective oral treatment strategies that have been widely accepted by both health-care professionals and the lay public. This article examines the correlation between the available biochemical and clinical data for the PDE 5 inhibitors sildenafil (Viagra), tadalafil (Cialis) and vardenafil (Levitra). PMID- 14551573 TI - Vasoconstriction, RhoA/Rho-kinase and the erectile response. AB - Recent studies have suggested that contraction of the smooth muscle in the cavernosal arterioles and in the walls of the cavernosal sinuses is maintained by the RhoA/Rho-kinase signaling pathway. However, this contraction activity must be overcome to permit the vasorelaxation essential for erection. We postulate that nitric oxide (NO) causes erection primarily by inhibiting the RhoA/Rho-kinase pathway. The following will discuss evidence in support of the important role of Rho-kinase-mediated vasoconstriction in the nonerect penis and how NO overrides this Rho-kinase-mediated vasoconstriction to permit vasodilation and erection. PMID- 14551574 TI - Update on central function relevant to sex: remodeling the basis of drug treatments for sex and the brain. AB - Sexual function draws on a complex network of peripheral and central neural pathways. The standard focus on erectile difficulties and peripheral therapies has been highly successful clinically but there are many unresolved issues in men and newly discussed issues in women that will likely benefit from improved understanding of the central nervous system and sexual function. The spectrum of future therapies, based on evolving central neurophysiological understanding, will include the management of problems related to orgasm, ejaculation, desire, motivation, anxiety, and pleasure. This new range of therapies will employ old and new neurochemicals and pathways singly or in combination. The capability of hormones to modulate many of the sexual pathways will also contribute to the rise of multiagent therapy. The expanded understanding, in combination with enhanced imaging technologies, will renew the role of diagnosis and cause-specific treatment. PMID- 14551575 TI - Frontiers in gene therapy for erectile dysfunction. AB - Complete sequencing of the human genome has made possible a new age of molecular medicine. The utilization of sophisticated genomic technologies has important implications to the understanding, diagnosis and treatment of erectile dysfunction. This report will review one aspect of the impact of the genomic revolution on urology, to wit, the preclinical evidence emerging from several laboratories indicating that gene therapy for erectile dysfunction may well provide the first safe and effective application of gene therapy to the treatment of human smooth muscle disease. The molecular targets explored thus far have concentrated largely on manipulating various aspects of the nitric oxide/guanylate cyclase/cGMP system, although genetic modulation of growth factors, calcium sensitization mechanisms and potassium channel expression have also been explored. Cell-based gene therapy techniques are also being explored. The apparent preclinical success of virtually all of these gene-based strategies reflects the multifactorial nature of erectile disease as well as the numerous regulatory mechanisms available for restoring erectile capacity. While technical hurdles remain with respect to the choice of delivery vectors, molecular target validation and duration of efficacy, 'proof-of-concept' has clearly been documented. The ultimate goal of gene therapy is to provide a safe, effective and specific means for altering intracavernous pressure 'on demand', while simultaneously eliminating the necessity for other forms of therapy, and moreover, without altering resting penile function, or the physiology of other organ systems. It is in these arenas that the groundbreaking potential of gene transfer technology to the treatment of erectile dysfunction will be fully tested. In fact, the potential benefits of the application of gene transfer techniques to this important medical problem is just now beginning to be appreciated/recognized. PMID- 14551576 TI - The human sexuality education of physicians in North American medical schools. AB - Individuals seeking treatment for sexual problems frequently would like to turn to a source they consider knowledgeable and worthy of respect, their doctor. The objective was to assess how well the 125 schools of medicine in the United States and the 16 in Canada prepare physicians to diagnose and treat sexual problems. A prospective cohort study was carried out. The main outcome results were description of the medical educational experiences, teaching time, specific subject areas, clinical programs, clerkships, continuing education programs in the domain of human sexuality in North American medical schools. The results were as follows. There were 101 survey responses (71.6%) of a potential of 141 medical schools (74% of United States and 50% of Canadian medical schools). A total of 84 respondents (83.2%) for sexuality education used a lecture format. A single discipline was responsible for this teaching in 32 (31.7%) schools, but a multidisciplinary team was responsible in 64 (63.4%) schools (five schools failed to respond to the question). The majority (54.1%) of the schools provided 3-10 h of education. Causes of sexual dysfunction (94.1%), its treatment (85.2%) altered sexual identification (79.2%) and issues of sexuality in illness or disability (69.3%) were included in the curriculum of 96 respondents. Only 43 (42.6%) schools offered clinical programs, which included a focus on treating patients with sexual problems and dysfunctions, and 56 (55.5%) provided the students in their clerkships with supervision in dealing with sexual issues. In conclusion, expansion of human sexuality education in medical schools may be necessary to meet the public demand of an informed health provider. PMID- 14551577 TI - Sexual health innovations in undergraduate medical education. AB - Recent national and global initiatives have drawn attention to the importance of sexual health to individuals' well-being. These initiatives advocate enhancement of efforts to address this under-represented topic in health professions curricula. University of Massachusetts Medical School (UMMS) has undertaken a comprehensive effort to develop an integrated curriculum in sexual health. The UMMS project draws upon the expertise of a multidisciplinary faculty of clinicians, basic scientists, a medical ethicist, and educators. This article describes the project's genesis and development at UMMS, and reports on three innovations in sexual health education implemented as part of this endeavor. PMID- 14551578 TI - A comprehensive approach to enhancing sexual health education in the Case Western Reserve University School of Medicine. AB - We report on the Sexual Health Curriculum Enhancement project at Case Western Reserve University School of Medicine. Using a US dollars 100000 grant from Pfizer Pharmaceuticals, Inc., we have developed and are in the process of implementing a comprehensive, cross-disciplinary and innovative curriculum that is based on three primary objectives for teaching sexual health: attitude change, behavior change, and knowledge acquisition. Five general strategies to incorporate specific sexual health content into the medical school curriculum have been implemented: (1). Faculty Development; (2). Additional Didactics; (3). Cased-Based Learning; (4). Testing and Assessment; and (5). Electronic (Computer/Web-Based Enhancements). PMID- 14551579 TI - Medical students' perceptions of sexual health issues prior to a curriculum enhancement. AB - The objectives were to compare first, second and third year medical students on self-perceived sexual health knowledge, comfort in addressing sexual health problems, and attitudes towards the importance of addressing sexual health issues with patients as part of a sexual health medical curriculum enhancement project. A paper-and-pencil questionnaire survey was designed and administered to first and second year medical students at the start of the fall semester, resulting in high participation rates for both years (98% and 86%, respectively). Third year students were surveyed through an on-line version of the questionnaire yielding a lower response rate (52%). Multivariate statistical analyses were used to compare knowledge, comfort and attitudes by year in medical school. Results were as follows: As might be expected, sexual health knowledge and comfort in addressing sexual health problems increased linearly from first to third year (P<0.01) for all questions. Unexpectedly, second year students had significantly higher scores on questions assessing attitudes towards the importance of addressing sexual health issues than either first or third year students (P<0.001). Female medical students reported that addressing sexual health issues with patients was significantly more important than did male medical students; however, male students reported higher levels of self-reported knowledge and comforting related to sexual health issues than did female students in a number of areas. In conclusion, knowledge gained from this survey was used to finalize the design of an enhanced, integrated curriculum on sexual health for medical students. Further investigation of gender differences related to training medical students in this area is suggested. PMID- 14551580 TI - Sex coaching for physicians: combination treatment for patient and partner. AB - Physicians dealing with sexual dysfunction (SD) must consider the psychological and behavioral aspects of their patient's diagnosis and management, as well as organic causes and risk factors. Integrating sex therapy and other psychological techniques into their office practice will improve effectiveness in treating SD. This presentation provides information about the psychological forces of patient and partner resistance, which impact patient compliance and sex lives beyond organic illness and mere performance anxiety. Four key areas are reviewed: (1). 'Sex coaching for physicians' uses the 'Cornell Model' for conceptualizing and treating SD. A 5-min 'sex status,' manages 'time crunch' by rapidly identifying common causes of sexual dysfunction (insufficient stimulation, depression, etc). (2). Augmenting pharmacotherapy with sex therapy when treating erectile dysfunction (ED) specifically, or SD generally is stressed. Sex therapy is useful as a monotherapy or an adjunctive treatment and is often the 'combination therapy' of choice when treating SD. The following therapeutic integrations are highlighted: modifying patient's initial expectations; sexual pharmaceuticals use as a therapeutic probe; 'follow-up' to manage noncompliance and improve outcome; relapse prevention. (3). Issues specific to the role of the partner of the ED patient are described. The physician must appreciate the role of couple's issues in causing and/or exacerbating the ED and the impact of the ED on the patient/partner relationship. Successful treatment requires a supportive available sexual partner, yet partner cooperation may be independent of partner attendance during the office visit. Preliminary data from a survey of SMSNA members practice patterns, regarding partner issues, is presented and discussed. The importance of evoking partner support and cooperation independent of actual attendance during office visits is emphasized. (4). Finally, the need for more patient and partner educational materials to assist the physician in overcoming a patient/partner's emotional barriers to sexual success in a time efficient manner are discussed. PMID- 14551581 TI - A framework for the present and future development of experimental models of female sexual dysfunction. AB - Female sexual dysfunction (FSD) is currently categorized according to disorders of (i). desire, (ii). arousal, (iii). orgasm and (iv). sexual pain. The advancement of research defining the physiological, pathophysiological and psychological mechanisms of these disorders, and to develop treatments for FSD, has been hampered by the paucity of experimental paradigms and animal models. It may be that animal models of FSD are best suited to address arousal disorders that include persistent or routine inability to attain or maintain genital lubrication or engorgement. Although still limited in scope, experimental models of FSD have involved a range of in vitro to in vivo methodologies. Specifically, the in vitro and in situ models include vaginal or clitoral smooth muscle preparations, histological evaluation and vaginal blood flow assessments. Previously, in vivo studies of sexual responses focussed on behavioral paradigms involving lordotic posturing and receptivity, as well as indices of motivation using a dual chamber pacing method. Recently, a new model of female sexual arousal was developed using pharmacological CNS stimulation; responses that were found to be sensitive to cardiovascular status, aging and hormonal conditions. It is important that a wide variety of animal models continue to be developed to reflect the multifactorial basis of the condition. PMID- 14551582 TI - Priapism pathophysiology: clues to prevention. AB - Priapism, in which penile erection persists in the absence of sexual excitation, is an enigmatic yet devastating erectile disorder. Current endeavors to manage the disorder suffer from a poor fundamental knowledge of the etiology and pathogenesis of priapism. These endeavors have remained essentially reactive, which commonly fail to avert its pathological consequences of erectile tissue damage and erectile disability, not to mention its psychological toll. The role of preventative management seems paramount with respect to priapism. As a prerequisite to formulating prevention strategies, gaining understanding of its pathogenic features and likely pathophysiologic mechanisms is viewed to be quite important. This review combined an analysis of clinicopathologic reports as well as a summary of clinical and basic science investigations on the subject to date. These assessments support the basic classification of priapism into low-flow (ischemic) and high-flow (nonischemic) hemodynamic categories, resulting from venous outflow occlusion and unregulated arterial overflow of the penis, respectively. In addition, consistent with the hypothesis that dysregulative physiology of penile erection accounts for some presentations of priapism, several plausible molecular mechanisms influencing the functional state of the erectile tissue are discussed. Current progress in the field suggests prevention possibilities using androgenic suppressive therapy, adrenergic agonist therapies, and effectors of the nitric oxide-dependent erection regulatory pathway in the penis. New ideas for prevention may emerge from targeting molecular mechanisms involved in regulating erectile tissue function. PMID- 14551583 TI - The treatment of priapism--when and how? AB - Priapism is a condition first described by Tripe in 1845. It has been defined as a pathological condition of penile erection that persists beyond or is unrelated to sexual stimulation. Two variants of priapism have been well described. The ischemic priapism (also known as low-flow priapism) and nonischemic priapism (or high flow priapism) have unique and distinct causes. It is important to distinguish these two conditions as the treatment for each is different. This review will focus on the two types of priapism and the appropriate diagnostic work-up for each. As well, the medical and surgical treatment options for these two conditions will be described in detail. A third entity known as stuttering priapism will also be discussed as will its unique treatment alternatives. PMID- 14551584 TI - History, epidemiology, and clinical presentation of Peyronie's disease. AB - Peyronie's disease is a localized connective tissue disorder that involves the tunica albuginea of the penis. The formation of fibrotic plaques in the tunica albuginea and surrounding cavernosal tissue alters penile anatomy and can cause different degrees of bending and narrowing, as well as penile pain and erectile dysfunction. Although long recognized as an important clinical entity of the male genitalia, the etiology of this disease has remained poorly understood. Recent studies have demonstrated that this disorder may be more prevalent than previously reported. The following review will examine the history, epidemiology, and clinical presentation of Peyronie's disease. PMID- 14551585 TI - Expanding the paradigm for plaque development in Peyronie's disease. AB - Despite centuries of recognition, Peyronie's disease remains somewhat of an enigma. Conventional wisdom suggests that trauma to the erect or semierect penis is the inciting event that sets off a cascade of events at the cellular level that results in localized fibrosis of the tunica albuginea. However, many questions remain unanswered at this juncture among the most important of which are why do so few men manifest this condition? Why is there such an ethnic predilection? What are the cofactors that along with penile trauma lead to plaque development. Historically, cytokine overexpression, autoimmune and genetic factors have been cited as contributors. This treatise endeavors to conduct an evidence-based assessment of the literature as it pertains to the pathophysiology of Peyronie's disease. Furthermore, an effort is made to evaluate contemporary literature pertaining to novel concepts in Peyronie's disease pathogenesis including NOS alterations, free radical generation, microarray analysis, pathogen involvement and animal model development. In conclusion, it is likely that in the near future we will see dramatic developments in our understanding of this condition. PMID- 14551586 TI - Establishing a standardized evaluation of the man with Peyronie's disease. AB - The goal of this article is to encourage development of a standardized approach to the evaluation of the man with Peyronie's disease. An additional aim is to identify those subjective and objective measures of change occurring as a result of treatment which would allow uniform reporting of treatment outcome. Systematic review of current clinical parameters used in assessing the man with Peyronie's disease including demographic data, physical findings, objective measures of penile deformity and erectile function as well as questionnaires employed to gain more reliable subjective data. As there is no accepted standard approach or previously published analysis of existing approaches, a survey of 68 recently published articles is included to query current outcome reporting on Peyronie's disease therapy. Results were as follows. An algorithm is suggested as an initial format for evaluation and outcome analysis. The pros and cons of subjective and objective measures of penile deformity is presented and a standardized questionnaire and penile duplex ultrasound report form is included for current use. The literature survey revealed the broad unscientific approach to outcome reporting making assessment of treatment quite difficult. In conclusion, it is clear that it is time to establish a standardized and globally acceptable evaluation and outcome reporting algorithm. There is interest to develop and validate a specific Peyronie's disease index questionnaire. In the meantime, this article presents specific recommendations to obtaining meaningful objective measures and methods to report them. PMID- 14551587 TI - Review of current nonsurgical management of Peyronie's disease. AB - The current nonsurgical treatment options for men with Peyronie's disease is reviewed. The treatments currently employed and published in the English literature are discussed. A wide variety of nonsurgical treatment options are available to the practicing physician, including oral and topical medications, intralesional injection therapy, as well as employing external energy sources to drive medicine into the tunica albuginea by iontophoresis or direct stimulation of plaque change by shock wave therapy. Nonsurgical treatment of Peyronie's disease clearly has a place in the armamentarium of the practicing urologist. Oral therapy appears to have little therapeutic benefit. Injection and topical approaches deserve further attention. Yet, no single treatment stands out as the most effective remedy for all men with Peyronie's disease. Further controlled, large-scale studies are necessary to establish the benefits of these nonsurgical approaches. In the meantime, combination therapy appears to make sense in the nonsurgical treatment of men with Peyronie's disease. PMID- 14551588 TI - Surgical approaches for advanced Peyronie's disease patients. AB - Surgical therapy for Peyronie's disease (PD) is reserved for patients with severe penile deformity that fails to improve with medical treatment and impedes sexual intercourse. The surgical treatment of PD consists of either correction of the penile deformity or insertion of a penile prosthesis in patients with concomitant erectile dysfunction (ED). Plaque incision/excision combined with grafting procedures or plication techniques are suitable in Peyronie's patients having an adequate penile vascular supply. When patients with Peyronie's disease have both penile deformity and ED, penile prosthesis implantation with or without excision or incision of the tunica is the current standard of care. PMID- 14551589 TI - Reimplantation of inflatable penile prosthesis into scarred corporeal bodies. AB - Reimplantation of an inflatable three-piece prosthesis in a patient with severe corporeal fibrosis represents a formidable surgical challenge. In the past, a variety of techniques have been offered to facilitate implantation in these difficult patients including the use of instruments to produce a channel in which cylinders can be placed and choosing multiple surgical sites, including the use of surgical sites that have not previously been used. The author of this communication describes newer tools and techniques to enhance placement of an inflatable device in patients with severe fibrosis. This includes the use of specially designed cavernotomes for dilating fibrotic corpora, the use of downsized prosthetic cylinders, alternative procedures to fix cylinders in the face of perforation as opposed to primary closure of the perforation, and replacing the original cylinders after 1 y after the modified cylinders have served as tissue expanders. Details of these newer techniques are presented by an experienced surgeon. PMID- 14551590 TI - Use of penile implants in the constructed neophallus. AB - Creation of a neophallus in cases of loss of the penis or female to male trans sexual is a challenge. The forearm flap is the most popular method used. A penile implant may be placed at the original procedure or at a later date to provide support for intercourse. The hydraulic nondistally expanding types provide the best support with less chance of distal erosion. Covering the device with cadaver pericardium or porcine small intestinal submucosa has been helpful in creating a neocorpus cavernosum. PMID- 14551591 TI - Cylinder sizing: less is more. AB - In penile prosthesis implantation traditional cylinder-sizing techniques frequently result in penile cylinders that are too long for the corpora cavernosa. This can result in premature cylinder wear leading to cylinder aneurysm or fluid loss. Additionally, in the case of the length expanding Ultrex penile prosthesis, cylinders that are too long can result in an S-shaped cylinder deformity. We describe a cylinder measurement technique that avoids the problem of oversizing. PMID- 14551592 TI - Surgical approaches for penile prosthesis implantation: penoscrotal vs infrapubic. AB - Surgical approaches for implantation of three-piece inflatable penile prostheses include the infrapubic and the penoscrotal. The penoscrotal approach avoids possible injury to the dorsal sensory nerves, provides easier and more complete corporeal exposure, and allows the pump to be anchored in the scrotal pouch. A variant of the penoscrotal approach, the transverse penoscrotal incision, is described. PMID- 14551593 TI - Three-piece inflatable penile prosthesis components (surgical pearls on reservoirs, pumps, and rear-tip extenders). AB - Erectile dysfunction (ED) is a major quality-of-life disorder that affects greater than 30 million American men each year. Since the introduction of Viagra, more and more men are presenting with ED that is not responding to oral or local therapies. Penile prosthesis implantation is recognized as the most effective, although most invasive, ED therapy with a high satisfaction rate. With an aging population both aware of and requesting therapy for ED, the number of penile prosthesis implantations will remain stable, especially at centers specializing in ED treatment. Therefore, a better understanding of the techniques used for placement of the penile prosthesis is necessary. The following review will discuss the techniques and a few clinical pearls in the placements of inflatable penile prostheses. PMID- 14551594 TI - Diagnosis, treatment and prevention of penile prosthesis infection. AB - The implantation of inflatable penile prostheses for the treatment of erectile dysfunction continues to be widely practiced in the United States and internationally. As third-line therapy for erectile dysfunction, the numbers of implants continue to rise as the population of men treated for erectile dysfunction increases. Complications of penile prosthesis implantation continued to decline as mechanical malfunctions have decreased as a result of re engineering inflatable penile prostheses. Inflatable penile prostheses from both available vendors continue to be reliable, effective methods for restoring erectile function with high satisfaction rates. The most troublesome complication of these prostheses, however, is not mechanical but rather that of prosthesis infection. Prosthesis infections may result in further surgery, loss of penile tissue, and even the inability to replace penile prosthesis. While standard sterile technique perioperative antibiotics and careful surgical procedures continue to be the cornerstone of penile prosthesis infection avoidance, newer designs of penile prostheses for antibiotic coating have resulted in an improvement in the prevalence and incidents of penile prosthesis infection. For those patients in whom penile prostheses become infected despite adequate prophylaxis, newer techniques of salvage have demonstrated increasing success. Once and still the most dreaded complication of penile prosthesis implantation, prothesis infections can now be avoided by perioperative preparation and antibiotics as well as antibiotic-coated penile prostheses. Treatment of penile prosthesis infections once associated with severe loss of function can often be successful with modern salvage techniques. Implanting urologists must be familiar prophylaxis, avoidance, and treatment of penile prosthesis infections. PMID- 14551595 TI - Treatment alternatives for the infected penile implant. AB - Infection associated with placement of a penile prosthesis is rare. When it does occur, the implant and all associated foreign material should be removed. The surgeon has the option of leaving the device out, allowing the wound to heal, and returning at a future date for prosthesis replacement if the patient wishes, or cleansing the wound and replacing the implant at the same procedure, termed a salvage procedure. The success rate of 'salvage' in this series of 101 patients was 84%. This approach is less likely to succeed when the infection is manifest soon (weeks) after the placement procedure, is accompanied by extensive cellulitis and is caused by virulent organisms. Relative contraindications to salvage include, sepsis, ketoacidosis, penile necrosis, and bilateral urethral erosion of the cylinders. PMID- 14551596 TI - Bacterial biofilms and implantable prosthetic devices. AB - Erectile dysfunction afflicts millions of men. A group of patients with advanced degeneration of their erectile tissue do not respond to pharmacological therapy, and surgical prosthetic reconstruction represents an attractive and highly satisfying alternative. Yet many men are unwilling to take this step due to fear of infection. Implanted prosthetic devices are at risk for infection because they provide a platform for the development of a bacterial biofilm, an organized bacterial colony that grows on the surface of the implanted material. The biofilm is resistant to all efforts to eradicate it short of removal of the foreign material. Bacteria may attach to the surface of the foreign material by surface charge attraction, hydrophilic/hydrophobic interactions, and by specific attachment by fimbrae. Growth, colonization, and maturation follow bacterial attachment. A mature biofilm is composed of three layers: a linking film binding the biofilm to the surface; a base film made up of a compact layer of bacteria; and a surface film from which free-floating bacteria can arise and spread. The majority of the surface layer is made up of exopolysaccharide matrix. Bacteria deep within the biofilm matrix live in a protected environment; diffusion of antibiotics is difficult, low oxygen tension leads to a lower bacterial metabolic rate rendering the bacteria functionally resistant to high levels of antibiotics. Effective strategies to reduce prosthetic infection levels must rely on the prevention of biofilm formation through surface modification. Possible mechanisms include the addition of antimicrobials to the surface of the device, or chemical modifications, which reduces bacterial attachment. PMID- 14551597 TI - Utility of magnetic resonance imaging in evaluating inflatable penile prosthesis malfunction and complaints. AB - The introduction of oral pharmacotherapy dramatically changed practice patterns in erectile dysfunction. The direct effect was to decrease the numbers of patients seeking penile implants; an indirect effect has been the changing ratio of new, to re-do operations. Patients seeking replacement inflatable penile prosthesis (IPP) surgery pose challenges both in diagnosis and management; in very select cases preoperative MRI can be useful in the evaluation of the patient's complaint and in planning operative management. Imaging can supplement the physical exam; in certain cases imaging will reveal: crural herniation, corporal distortion, corporal fibrosis, and hardware migration. Inflatable penile prosthesis has a high rate of satisfaction for patients (89%) and partners (70%). The principal reasons for dissatisfaction are penile shortening, pain and frustration with reoperation. If pain is not due to infection, it may be secondary to malpositioning, improper sizing, cylinder cross-overs, or herniation. After one or more reoperations, penile distortion may occur secondary to tunica albugenia thinning or fibrosis. A series of cases will be presented to highlight the utility of MRI for IPP problems, define normal appearance and abnormal appearance of penile hardware. Careful collaboration between the Urologist and Radiologist in the review and final dictation of these cases is warranted to properly document problems and to plan device replacement/penile reconstruction. PMID- 14551598 TI - Simultaneous analysis of interleukin-10 gene microsatellites and single nucleotide polymorphisms in parallel with tumour necrosis factor and interferon gamma short tandem repeats by fluorescence-based polymerase chain reaction. AB - Different cytokine genotypes exist in the population, for example, as a result of selective pressure of infectious diseases. It may be that specific cytokine genotypes that are beneficial by creating a 'proinflammatory' phenotype predispose to severe inflammatory disease with worse clinical outcome. There is individual variation in the production of certain cytokines in relation to their genotypes. IL-10, IFN-gamma and TNF-alpha are key components in the regulation of immune responses and the balance of their expression levels is predictive in certain diseases. To describe cytokine genotypes, a one-tube PCR reaction was developed to analyse simultaneously DNA sequence variations of cytokine genes IL 10, IFN-gamma, and TNF. This multiplex PCR approach was used to provide genotypic data for two geographically independent donor groups from Germany and Gabon. Significant differences were obtained for the majority of sequence variations comparing both populations. However, the SNPs within the 5'-flanking region of the IL-10 gene at position -1087 and -6208 are comparable in their genic and genotypic behaviour. Comparing allelic and genotypic disequilibrium between pairs of loci revealed different association patterns for both populations according to the geographical polymorphism. This assay may improve immunogenetic studies in disease, characterized by disbalanced IL-10, IFN-gamma and TNF-alpha expression. PMID- 14551599 TI - Testing the possible negative association of type 1 diabetes and atopic disease by analysis of the interleukin 4 receptor gene. AB - Variations in the interleukin 4 receptor A (IL4RA) gene have been reported to be associated with atopy, asthma, and allergy, which may occur less frequently in subjects with type 1 diabetes (T1D). Since atopy shows a humoral immune reactivity pattern, and T1D results from a cellular (T lymphocyte) response, we hypothesised that alleles predisposing to atopy could be protective for T1D and transmitted less often than the expected 50% from heterozygous parents to offspring with T1D. We genotyped seven exonic single nucleotide polymorphisms (SNPs) and the -3223 C>T SNP in the putative promoter region of IL4RA in up to 3475 T1D families, including 1244 Finnish T1D families. Only the -3223 C>T SNP showed evidence of negative association (P=0.014). There was some evidence for an interaction between -3233 C>T and the T1D locus IDDM2 in the insulin gene region (P=0.001 in the combined and P=0.02 in the Finnish data set). We, therefore, cannot rule out a genetic effect of IL4RA in T1D, but it is not a major one. PMID- 14551600 TI - Complex haplotypic structure of the central MHC region flanking TNF in a West African population. AB - TNF polymorphisms have been associated with susceptibility to malaria and other infectious and inflammatory conditions. We investigated a sample of 150 West African chromosomes to determine linkage disequilibrium (LD) between 25 SNP markers located in an 80 kb segment of the MHC Class III region encompassing TNF and eight neighbouring genes. We observed 45 haplotypes, and 22 of them comprise 80% of the sample. The pattern of LD is remarkably patchy, such that many markers show no LD with adjacent markers but high LD with markers that are much further away. We introduce a method of examining the implications of LD data for disease association studies based on sample size considerations: this shows that certain TNF polymorphisms would be likely to yield positive associations if the true disease allele resided in LTA or BAT1. We conclude that detailed marker maps are needed to resolve the causal origin of disease associations observed at the TNF locus. PMID- 14551601 TI - Macrophage migration inhibitory factor (MIF) gene polymorphism is associated with susceptibility to but not severity of inflammatory polyarthritis. AB - The aim of the study was to investigate whether polymorphisms of macrophage migration inhibitory factor (MIF) determine susceptibility to or severity of inflammatory polyarthritis (IP). Genotypes for a single-nucleotide polymorphism (MIF-173*G/C) and a tetranucleotide (CATT)(n) repeat mapping to the promoter region of the MIF gene were compared between UK Caucasian IP cases (n=438) and controls (n=343). Both polymorphisms were also investigated for association with features of disease activity and severity at baseline and by 5 years. The MIF 173*C allele (OR 1.7, 95% CI 1.3-2.4, P=1.8 x 10(-4)) and the CATT(7) allele (OR 1.5, 95% CI 1.0-2.1, P=0.02) were found to be associated with increased susceptibility to IP. Furthermore, presence of the haplotype containing both associated polymorphisms was associated with a three-fold increase risk of developing IP. No association with disease severity or activity either at baseline or by 5 years was detected for either of the promoter polymorphisms studied. In conclusion, MIF is a susceptibility gene for the development of IP. The same alleles previously reported to be associated with susceptibility to juvenile idiopathic arthritis account for the increased risk. The promoter polymorphisms of MIF, investigated in this study, do not influence the severity of disease outcome by 5 years. PMID- 14551602 TI - Genes of the LMP/TAP cluster are associated with the human autoimmune disease vitiligo. AB - Genes within the class II region of the major histocompatibility complex (MHC), including genes involved in antigen processing and presentation, have been reported to be associated with several autoimmune diseases. We report here that the LMP/TAP gene region is significantly associated with vitiligo, a disorder in which biochemical defects and/or autoimmune destruction cause melanocyte loss and resulting skin depigmentation. Case/control analyses revealed genetic association of vitiligo in Caucasian patients with an early age of onset with the transporter associated with antigen processing-1 (TAP1) gene. A family-based association method revealed biased transmission of specific alleles from heterozygous parents to affected offspring for the TAP1 gene, as well as for the closely linked LMP2 and LMP7 genes encoding subunits of the immunoproteasome. No association with vitiligo was found for the MECL1 gene, which encodes a third immunoproteasome subunit and is unlinked to the MHC class II region. These results suggest a possible role for the MHC class I antigen processing and/or presentation pathway in the antimelanocyte autoimmune response involved in vitiligo pathogenesis. PMID- 14551603 TI - MICA, HLA-B haplotypic variation in five population groups of sub-Saharan African ancestry. AB - The human major histocompatibility complex (MHC) class I chain-related gene A (MICA), located 46 kb centromeric to HLA-B, encodes a stress-inducible protein, which is a ligand for the NKG2D receptor. In addition to its primary role in immune surveillance, data suggest that MICA is involved in the immune response to transplants and in susceptibility to some diseases. In this study, 152 subjects from the Yoruba (n=74), Efik (n=32), and Igbo (n=46) tribes of southern Nigeria, 39 nationwide African-American stem cell donors, and 60 African-American individuals residing in the metropolitan Boston area were studied for MICA, HLA-B allelic variation, haplotypic diversity, and linkage disequilibrium (LD). MICA and HLA-B exhibited a high degree of genetic diversity among the populations studied. In particular, MICA allele and HLA-B-MICA haplotype frequencies and LD in the Efik and Igbo tribes were significantly different from the other study groups. HLA-B and MICA loci demonstrated significant global LD in all five populations (P-values &<0.00001). LD also varied in a haplotype-specific manner. A novel MICA allele was detected in the Boston population. These findings are important from an anthropologic perspective, and will inform future HLA-linked disease association studies in related ethnic groups of African-derived ancestry. PMID- 14551604 TI - Haplotypic relationship between SNP and microsatellite markers at the NOS2A locus in two populations. AB - The density of genetic markers required for successful association mapping of complex diseases depends on linkage disequilibrium (LD) between non-functional markers and functional variants. The haplotypic relationship between stable markers and potentially unstable but highly informative markers (e.g. microsatellites) indicates that LD might be maintained over considerable genetic distance in non-African populations, supporting the use of such 'mixed marker haplotypes' in LD-based mapping, and allowing inferences to be drawn about human origins. We investigated sequence variation in the proximal 2.6 kb of the inducible nitric oxide synthase (NOS2A) promoter and the relationship between SNP haplotypes and a pentanucleotide microsatellite (the 'NOS2A(-2.6) microsatellite') in Gambians and UK Caucasians. UK Caucasians exhibited a subset of sequence diversity observed in Gambians, sharing four of 11 SNPs and a similar haplotypic structure. Five SNPs were found in the sequence of interspersed repetitive DNA elements. In both populations, there was dramatic loss of LD between SNP haplotypes and microsatellite alleles across a very short physical distance, suggesting a high intrinsic mutation rate of the NOS2A(-2.6) microsatellite, the SNP haplotypes are relatively ancient, or that this was a region of frequent recombination. Understanding locus- and population-specific LD is essential when designing and interpreting genetic association studies. PMID- 14551605 TI - Association between polymorphisms in the SPINK5 gene and atopic dermatitis in the Japanese. AB - Atopy, which is characterized by increased levels of immunoglobulin E (IgE) against common environmental allergens, is considered the strongest predisposing factor for asthma and atopic dermatitis (AD). Mutations in the gene encoding serine protease inhibitor Kazal-type 5 (SPINK5) are responsible for Netherton syndrome, a rare skin disorder characterized by greatly elevated IgE levels with atopic manifestations. A recent study of Caucasian AD families showed that maternally derived alleles of the SPINK5 gene are associated with development of AD and asthma, suggesting the parent-of-origin effect for the development of atopic diseases in the SPINK5 gene. We studied the possible association of the SPINK5 gene for the development of atopic diseases by determining the genotypes of five polymorphisms in a Japanese population. Ttransmission disequilibrium tests revealed an association of SPINK5 polymorphisms with AD but not with asthma. Our data indicate that the SPINK5 gene is associated with AD across ethnicities. PMID- 14551606 TI - Intercellular adhesion molecule-1: a protective haplotype against multiple sclerosis. AB - Intercellular adhesion molecule-1 (ICAM-1) and its receptors are adhesion molecules that play a key role in the transmigration of inflammatory cells through the blood-brain barrier, one of the earliest events in multiple sclerosis (MS), which leads to demyelination in the central nervous system. To investigate the role of genes encoding ICAM-1 and its receptors, we used a strategy of genetic linkage and association in 439 case-parent MS families of French origin, well characterized according to HLA status and severity. We demonstrate that the genes encoding ICAM-1 receptors do not influence MS susceptibility or severity. ICAM-1 had a modest, but significant effect on MS genetic susceptibility, independent of HLA and disease severity. We observed a rare, and an as yet unreported, ICAM-1 gene haplotype defined by amino acids K469 and R241 that was never transmitted to patients suggesting a protective effect against MS in our population. PMID- 14551607 TI - Polymorphisms of innate immunity genes and susceptibility to lymphatic filariasis. AB - We examined 906 residents of an area of Papua New Guinea where bancroftian filariasis is endemic for genetic polymorphisms in three innate immunity genes suspected of contributing to susceptibility to infection and lymphatic pathology. Active infection was confirmed by the presence of blood-borne microfilariae and circulating filarial antigen in plasma. Disease was ascertained by physical examination for the presence of overt lymphedema (severe swelling of an arm or leg) or hydrocele. There was no association of infection status, lymphedema of an extremity, or hydrocele with chitotriosidase genotype (CHIT1). Polymorphisms of toll-like receptor-2 and toll-like receptor-4 genes (TLR4 A896G; TLR2 T2178A, G2258A) were not detected (N=200-625 individuals genotyped) except for two individuals heterozygous for a TLR2 mutation (C2029 T). These results indicate that a CHIT1 genotype associated previously with susceptibility to filariasis in residents of southern India and TLR2 and TLR4 polymorphisms do not correlate with infection status or disease phenotype in this Melanesian population. PMID- 14551608 TI - A single-nucleotide substitution from C to T at position -1055 in the IL-13 promoter is associated with protection from severe malaria in Thailand. AB - We examined a possible association of single-nucleotide polymorphisms (SNPs) in the promoters of IL-3, IL-4, and IL-13 genes on the 5q31-33, IL-3 -16T>C, IL-4 590T>C, and IL-13 -1055C>T, with severity of malaria in 361 adult malaria patients in Thailand. The IL-13 -1055T allele showed a significant association with protection from severe malaria (OR 0.51, 95% CI 0.32-0.80; P=0.0032 by the chi(2) test), while allele frequencies of IL-3 -16T>C and IL-4 -590T>C were not statistically different between mild and severe malaria patients. An IL-13 1055C>T has been reported to alter the regulation of IL-13 production. Thus, IL 13 -1055T may show resistance to severe malaria through the alteration of IL-13 production. PMID- 14551609 TI - Questionnaire research: an easy option? PMID- 14551610 TI - Overbrushing. PMID- 14551611 TI - Tooth notation. PMID- 14551612 TI - Tooth notation. PMID- 14551613 TI - Tooth notation. PMID- 14551614 TI - Clinical audit. PMID- 14551615 TI - Do-it-yourself dentistry. PMID- 14551622 TI - General medicine and surgery for dental practitioners Part 10: The paediatric patient. PMID- 14551623 TI - Prevention. Part 8: The use of pit and fissure sealants in preventing caries in the permanent dentition of children. AB - This paper reviews evidence concerning the use of pit and fissure sealants in preventing caries in the permanent dentition of children. While the evidence with respect to some sealant types and application techniques is incomplete, systematic reviews have clearly demonstrated that sealants are an effective preventive technology when used in high risk children, and that with proper application techniques long-term retention rates can be achieved. However, careful selection of patients and teeth for sealant placement is required to ensure cost-effectiveness. PMID- 14551631 TI - Peer review amongst restorative specialists on the quality of their communication with referring dental practitioners. AB - OBJECTIVE: A peer review study was carried out to assess the written communication between consultants and specialist registrars in restorative dentistry with the referring general dental practitioners. METHODS: Seven people took part in the study and each presented referral and reply letters for five patients whom they had seen for consultation. The referral letters were used for information only and were not used in the peer review process. Each participant inspected the referral and reply letters from the other six participants. The reply letters were anonymously peer reviewed by using a proforma containing agreed criteria in relation to appropriate factors to include in the reply letter. The reviewer also ranked the letter in relation to overall quality on a 1 10 point scale. RESULTS: It was found that the participants' letters generally conformed positively with the agreed criteria although there were some differences between individuals. There were particular problems identified in relation to tooth notation. Reply letters commonly used different forms of tooth notation to the referring practitioners. CONCLUSIONS: The ranking of the letters generally indicated that the participants' replies were judged to be favourable by their peers. There may be scope for continuing this study in relation to peer review by other groups of professionals, in particular practitioners in primary dental care. PMID- 14551632 TI - A cluster randomised, controlled trial of the value of dental health educators in general dental practice. AB - AIM: To test the effectiveness of dental health educators in general dental practice. OBJECTIVE: To evaluate the effectiveness and cost of primary care trusts seconding dental health educators free of charge to suitable general dental practices to provide dental health counselling to mothers of regularly attending pre-school children at risk to caries. METHOD: Two-cell, parallel group, cluster randomised, controlled clinical trial of two years' duration. CLINICAL SETTING: 30 general dental practices in North-West England. PARTICIPANTS: 269 mothers of 334 pre-school children. INTERVENTIONS: Those in the test group were given visits to a dental health educator over a 2-year period to counsel mothers of at-risk, pre-school children. The rest were held as a control. MAIN OUTCOME MEASURES: Caries prevalence of the children and dental health knowledge, attitudes and toothbrushing skills of the parents. The full costs of the exercise were kept throughout. The statistical analysis controlled for the clustering of children within practices. RESULTS: After 2 years, 271 (81%) children and 248 (92%) mothers remained in the study. There was an 18% difference in mean dmft between the groups in favour of the test group children but this was not statistically significant. At the end of the study there was an 18% difference in mean dmft between the groups in favour of the test group children but this was not statistically significant. No difference in plaque levels was found. The mothers in the test group were more knowledgeable, had better attitudes towards the dental health of their offspring and better toothbrushing skills than those in the control. Each 2-hour session to counsel ten parents cost pound 40. CONCLUSION: Primary care trusts should carefully consider the cost value of seconding dental health educators to counsel parents of regularly attending, at-risk, pre-school children when considering such an option. PMID- 14551633 TI - Can psychological models bridge the gap between clinical guidelines and clinicians' behaviour? A randomised controlled trial of an intervention to influence dentists' intention to implement evidence-based practice. AB - OBJECTIVE: The lag between publication of evidence for clinical practice and implementation by clinicians may be decades. Research using psychological models demonstrates that changing intention is very important in changing behaviour. This study examined an intervention (rehearsing alternative actions) to change dentists' intention to implement evidence-based practice (EBP) for third molar (TM) management. DESIGN: Randomised controlled trial / postal. SETTING: Primary care. SUBJECTS AND METHODS: Dentists were randomly selected from the Scottish Dental Practice Board Register, then randomly allocated to intervention or control groups, and sent a questionnaire. The intervention group listed management alternatives to TM extraction prior to their TM extraction intention, and the control group did not. Based on psychological models for reducing a behaviour's frequency (EBP is weighted against TM extraction), prior listing of alternatives should decrease extraction intention. MAIN OUTCOME MEASURES: Intention to extract TMs. RESULTS: A total of 99 dentists - 70 Males, 29 Females; mean age = 41.42 years (SD = 8.62) participated in the study. The intervention significantly influenced intention to extract TMs, as desired. Despite similar background and knowledge of management alternatives, participants in the intervention group had significantly lower intention to extract: control group mean (SD) = 0.39 (1.99); intervention group mean (SD) = -0.78 (1.89); mean difference (SE) = 1.17 (0.42); 95% confidence interval for the difference = 0.34 to 1.99. CONCLUSION: Results suggest this intervention, which successfully influenced a proximal predictor of behaviour pertinent to dental EBP, may result in improved EBP in a service-level trial. Basing implementation interventions and trial methodology on psychological models may effectively bridge the gap between clinical guidelines and practice. PMID- 14551651 TI - The operating room of the future: observations and commentary. AB - The Operating Room of the Future is a construct upon which to develop the next generation of operating environments for the patient, surgeon, and operating team. Analysis of the suite of visions for the Operating Room of the Future reveals a broad set of goals, with a clear overall solution to create a safe environment for high-quality healthcare. The vision, although planned for the future, is based upon iteratively improving and integrating current systems, both technology and process. This must become the Operating Room of Today, which will require the enormous efforts described. An alternative future of the operating room, based upon emergence of disruptive technologies, is also presented. PMID- 14551652 TI - Visualization trends: applications in the operating room. AB - Recent advances in visualization technology are being driven by two important trends: (1) continued increases in speed and function of hardware devices and (2) increasingly parallel, distributed, cooperative systems. The incorporation of fast, powerful devices into cooperative systems enables a complex interplay of sensors, displays, and computational components that can create a seamless, perceptually rich and flexible environment. Although these trends have fueled a number of advances in visualization research, the unique requirements of laparoscopy make direct, effective use of visualization technology as it is applied in other contexts extremely challenging. This article discusses promising new capabilities in visualization technology. The costs and tradeoffs create new challenges, which are addressed in some visualization applications, but must be carefully assessed in the context of the laparoscopic environment. Incorporating new visualization technology in a way that captures its benefits and meets stringent laparoscopic requirements will very likely precipitate an enormous surge forward in the capabilities of the surgical team and in the quality of patient care. PMID- 14551653 TI - FDA regulation of technology and surgical devices in the operating room. AB - Federal regulation of medical devices began in 1976 with the signing of the Medical Device Amendments to the Food, Drug and Cosmetic Act. For the purpose of regulating medical devices, the Food and Drug Administration is divided into various divisions and branches, including the Office of Device Evaluation. The evolution of the Food and Drug Administration's regulations of laparoscopic devices is described. Also described is the technology of laparoscopic surgical devices and how they are regulated by the Office of Device Evaluation. Trends towards the future of laparoscopic devices, and their regulation, are reviewed. PMID- 14551654 TI - Telemedicine in tomorrow's operating room: a natural fit. AB - The integration of telecommunications and information technologies in medicine, known as telemedicine, has traditionally been outside the operating room. However, with the advent of new technologies, smart systems, and advanced computing, adoption and adaptation of these technologies in the operating room has increased. The operating room has been a place of startling isolation. The operating room is an island within a health system that is only reached by a select few. The application of telemedicine in this inaccessible place offers a huge potential in a variety of ways. These include enhanced education for students, enhanced safety for the patient, enhanced knowledge of the events that occur-a sort of status monitoring of underlying infrastructures-and sharing of the operating environment for collaboration. The Operating Room of the Future will be linked to other islands of expertise by robust telemedicine capabilities, thereby eliminating the isolation. Telemedicine in tomorrow's operating room is a natural fit PMID- 14551655 TI - From the operating room of the present to the operating room of the future. Human factors lessons learned from the minimally invasive surgery revolution. AB - The minimally invasive surgical revolution has changed the way surgery is practiced. It has also helped surgical innovators to break the tethers that anchored the practice of surgery in an early 20th century operating room environment. To some in surgery, the Operating Room of the Future will be seen as a revolution but to others, an inevitable evolution of the changes ushered in by the adoption of minimally invasive surgery. Although minimally invasive surgery has conferred considerable advantages on the patient, it has imposed significant difficulties on the surgeon, which in turn, have impacted outcomes. These difficulties were primarily human factor in nature and were poorly understood by critical groups such as device manufacturers, surgeons, and surgery educators and trainers. This article details what these human factors were, how they related to the practice of minimally invasive surgery, and how they will impact on the practice of surgery in the Operating Room of the Future. Much of the technology for the Operating Room of the Future currently exists (eg, surgical robotics, virtual reality, and telemedicine). However, for it to function optimally it must be integrated in a fashion that takes on board the human factor strengths and limitations of the surgeon. These advanced technologies should then be harnessed to optimize surgical practice. In some cases, this will involve rethinking existing technologies (ie, three-dimensional camera systems), applying technologies that currently exist in a manner that is more systematic and better managed (ie, surgical robots and virtual reality), and a reconsideration of who should be applying these technologies for the practice of surgery in the 21st century. In all cases, there will be education and training implications for the practitioner. Lastly, there must be unequivocal demonstration that these changes bring about positive benefits for patients in terms of better outcomes and for surgeons in terms of ability and ease of doing their job. After the experiences of the last decade with minimally invasive surgery, the Operating Room of the Future should be seen as a well-grounded evolution, not a revolution. PMID- 14551656 TI - Lest we forget the surgeon. AB - Within the United States, the applications from medical graduates to general surgery residency programs declined by 30% since 1992, and it has been estimated that by 2005 only 5% of US medical graduates will opt for a professional career in surgery. Thus in the assessment of the projected needs for surgical practice for the new century, we must not overlook the surgical manpower issue. We have to ensure that the high-tech operating rooms of the future will be manned by surgeons with the right personality, attitudes, competence, and skills. Certain key issue have to be addressed if we are to achieve this essential objective. These include changes in health care systems, including the rapidly advancing technologically-dependent minimal access therapy procedures, changes in attitude and culture between doctors and patients, especially in relation to human error enacted during health care delivery with the abolition of the "shame and blame culture," changes in the selection and training of surgeons that have to take into account the reduced working week for residents, and appraisal systems that will ensure sustained competence of fully trained surgeons throughout their professional life. Since it is not possible to eliminate errors completely from clinical practice, we can improve the quality of medical and surgical care by adopting error-tolerant operating medical systems based on progress in cognitive psychology, human factors, and human reliability assessment. Error-tolerant operating medical systems should enable detection, reporting, and targeted reduction of errors. PMID- 14551657 TI - The operating room of the future: a view from Europe. AB - The Operating Room of the Future will be characterized by meticulous preoperative planning, full integration of the operating room into the general flow of information, more comprehensive intraoperative diagnostic imaging procedures, and the use of sophisticated visualization processes including augmented reality. Mechatronic support (partially autonomous robots) enhances safety and allows reduction of staff. Integrated operating room systems will allow the wide spectrum of new devices and functionalities to be easily controlled by the operating team. The Operating Room of the Future will no longer be isolated from the rest of the clinical endeavor. Intraoperative teleconsultation and telepresence will help to promote and teach safer evidence-based endoscopic therapeutic surgery. Traditional surgical intervention will expand its definitions by procedures via an interdisciplinary, cooperative approach that will replace the sequential therapeutic process of today. PMID- 14551659 TI - Rapid analysis of aromatic contaminants in water samples by means of laser ionization mass spectrometry. AB - Three different approaches to laser ionization mass spectrometric analysis of aromatic compounds in water samples are described and their performances are compared. Whereas the first two methods are based on direct laser desorption and subsequent laser ionization of either frozen or adsorbed samples in a time-of flight mass analyzer, the third performs laser ionization in a quadrupole ion trap into which the sample is transferred from a GC injector via a short piece of capillary tubing. For the laser-desorption method a detection limit in the 100 microg L(-1) range was determined for fluorene in frozen samples. The easier to handle analysis of adsorbed samples yielded sensitivities which were lower by about two orders of magnitude. As both direct techniques do not reach the sensitivity required for ultra trace analysis in water a preconcentration step in form of solid-phase microextraction was added before measurement using the laser ionization quadrupole ion-trap mass spectrometer. Sensitivity in the desired ng L(-1) range was easily achieved. PMID- 14551658 TI - Analysis of serotonin release from single neuron soma using capillary electrophoresis and laser-induced fluorescence with a pulsed deep-UV NeCu laser. AB - The use of capillary electrophoresis (CE) with laser-induced fluorescence excited by ultraviolet (UV) lasers in the range 200-300 nm has been restricted by the available wavelengths and expense of UV lasers. The integration of a NeCu deep UV laser operating at 248.6 nm with a single channel CE system with post-column sheath flow detection allows detection limits for serotonin and tryptophan of 3.9x10(-8) M and 4.5x10(-8) M respectively. Single cell analysis of serotonergic metacerebral cells from the sea slug Aplysia californica yields a value of 800+/ 85 fmol of serotonin in each cell soma. For the first time, serotonin is directly detected in electrically stimulated release from single metacerebral cell soma, with approximately 4% of the serotonin contained in the soma released from a semi intact preparation with a 2 min electrical stimulation. PMID- 14551660 TI - Elemental fingerprint analysis of barley (Hordeum vulgare) using inductively coupled plasma mass spectrometry, isotope-ratio mass spectrometry, and multivariate statistics. AB - Inductively coupled plasma mass spectrometry (ICP-MS) and isotope-ratio mass spectrometry (IR-MS) have been used to examine the multi-elemental composition and (15)N/(14)N and (13)C/(12)C isotope ratios of three spring barley (Hordeum vulgare) genotypes (Orthega, Barke, and Bartok) grown in three typical Danish agricultural soils (North Jutland, West Jutland, and East Zealand) differing in soil fertility. The aim of the study was to examine whether it was possible to generate a unique elemental fingerprint of individual barley genotypes irrespective of the elemental imprint plants had received from soils differing in fertility and agricultural practice. Multivariate statistics were used to analyze the elemental fingerprints of the barley genotypes at different times during a full growing season from early tillering to full maturity of the barley grains. Initially, 36 elements were analyzed in the plant samples but this number was subsequently reduced to 15 elements: B, Ba, C, Ca, Cu, Fe, K, Mg, Mn, N, Na, P, S, Sr, and Zn. These elements exceeded the limit of detection ( LOD) for all genotypes, soil types, and plant growth stages and for these elements the accuracy was better than 90% compared with apple leaf certified reference material (CRM). Principal component analysis (PCA) separated multi-elemental data in accordance with soil type when plants of similar physiological age were compared, whereas this separation disappeared if plants of all ages were compared simultaneously. Isotope ratios (delta(15)N) of plants also proved to be a highly accurate property for classification of samples according to soil type. In contrast, the differences in delta(13)C were too small to enable such classification. The differences in delta(15)N among soils were so pronounced that separation of samples according to the physiological age of plants became redundant. However, delta(15)N and the multi-elemental analysis revealed no differences between the three barley genotypes, indicating that the influence of soil chemistry and possibly also climate and agricultural practice was too large to allow an unique elemental fingerprint for the genotypes. This finding was substantiated by analyzing the multi-elemental composition of grain from two additional genotypes (Otira and Barthos) grown at the north and east locations, respectively. PCA showed not only that the elemental fingerprints of these two genotypes were similar to those of the others, but also that the soil in which the plant had been growing could be accurately predicted on the basis of the PCA scores from the genotypes Orthega, Barke, and Bartok. Similar conclusions could be drawn using delta(15)N data. PMID- 14551661 TI - Comparison of different fluorimetric HPLC methods for analysis of acidic polyether toxins in marine phytoplankton. AB - The human toxic syndrome, diarrhetic shellfish poisoning (DSP), is caused by polyether toxins that are present in bivalve molluscs but originate from some species of marine phytoplankton. During the last few years different HPLC methods with fluorescence detection (FLD) have been proposed for analysis of marine toxins, including polyether toxins, in shellfish and phytoplankton. Several derivatization reagents have been proposed in the literature, with the aim of converting the acidic DSP toxins into their corresponding fluorescent derivatives. In this work we report results obtained from HPLC-FLD analysis of extracts from phytoplankton, including Dinophysis spp.,harvested off the south west coast of Ireland. Three different reagents were used for fluorescent derivatization: 3-bromomethyl-6,7-dimethoxy-1-methyl-2(1H)-quinoxalinone (BrDMEQ), 9-chloromethylanthracene (CA), and "in situ" 9-anthracenyldiazomethane (ADAM). Derivatization was performed under conditions previously optimised. The DSP derivatives were cleaned using different SPE procedures then analysed by HPLC FLD. In this study, the use of BrDMEQ, CA, and "in situ" ADAM was compared in terms of sensitivity and selectivity. Evaluation of HPLC methods for analysis of DSP toxin derivatives was also conducted; the presence of okadaic acid (OA), dinophysistoxin-2 (DTX-2), and pectenotoxin-2 seco acids (PTX1SAs) was detected in the sample extracts studied. PMID- 14551662 TI - Human urine certified reference material CZ 6009: creatinine, styrene metabolites (mandelic acid and phenylglyoxylic acid). AB - The reference material was prepared by freeze-drying pooled urine samples obtained from healthy persons occupationally exposed to styrene. The concentrations of mandelic acid (MA), phenylglyoxylic acid (PGA), and hippuric acid (HA) in urine were determined by three modes of high-performance liquid chromatography (HPLC). For isochronous stability testing the urinary mandelic acid and phenylglyoxylic acid concentrations were followed over a 24-month period for a preliminary batch by use of HPLC. No changes of the concentration values were found. The creatinine concentration was stable for more than five years. Standard Reference Material NIST 914a Creatinine was used for traceability purposes for creatinine. Pure chemicals MA and PGA were used for traceability purposes. Control material ClinChek-Urine Control (Recipe) was analyzed simultaneously. The mean values of MA and PGA compare well with the means and fall within the control range of control samples. Results from homogeneity, stability, and traceability testing were evaluated using the statistical program ANOVA. The certified values and their uncertainties were evaluated from the results of interlaboratory comparisons, and homogeneity and stability tests. The values are unweighed arithmetical averages of accepted results and their uncertainties are combined uncertainties (coverage factor=1). PMID- 14551663 TI - Measurement of isotope ratios on transient signals by MC-ICP-MS. AB - Precise and accurate isotope ratio measurements are an important task in many applications such as isotope-dilution mass spectrometry, bioavailability studies, or the determination of isotope variations in geological or nuclear samples. The technique of MC-ICP-MS has attracted much attention because it permits the precise measurement of isotope compositions for a wide range of elements combined with excellent detection limits due to high ionisation efficiencies. However, the results are based mainly on measurements using continuous sample introduction. In the present study the determination of isotope ratios on various transient signals with a time duration of 30 to 60 s has been achieved by coupling high performance liquid chromatography to a multicollector inductively coupled plasma mass spectrometer. In order to investigate the origin of ratio drifts across the transient signals for this hyphenated technique, measurements with the same standard solutions were also carried out using a flow-injection device for sample introduction. As a result of this application it could be concluded that the main source of the bias in the measured isotope ratios is within the ICP-MS instead of fractionation effects on the chromatographic column material. Preliminary studies on short transient signals of gaseous samples (dry plasma) showed a reverse fractionation effect compared with wet plasma conditions (flow injection and HPLC). PMID- 14551664 TI - ICP-MS measurements of lead isotopic ratios in soils heavily contaminated by lead smelting: tracing the sources of pollution. AB - The Pb isotopic composition (206Pb/207Pb and 208Pb/206Pb) in smelter-impacted soils was measured using a quadrupole-based ICP-MS. Four forest/tilled soil profiles were sampled according to the distance from the lead smelter in Pribram (Czech Republic), prevailing wind direction, geological background and soil type. The results were compared with the Pb isotopic composition of bedrocks and waste materials from Pb metallurgy (smelting slags, air-pollution-control residues). The isotopic composition of soils confirms the predominant role of metallurgy on the general pollution in the area. The highly contaminated soils from the vicinity of the smelter contain up to 35,300 mg Pb kg(-1) and exhibit an isotopic composition close to that of car battery processing (206Pb/207Pb up to 1.177). A coupled concentration/isotopic study of soil profiles showed that the smelter induced pollution had penetrated even to the mineral soil horizons, indicating an important vertical mobility of Pb contaminant within the soil profile. The calculated downward penetration rate of Pb in soils ranges from 0.3 to 0.36 cm year(-1). PMID- 14551665 TI - Use of electrospray ionization mass spectrometry for the investigation of radical cation chain reactions in solution: detection of transient radical cations. AB - Electrospray ionization mass spectrometry (ESI-MS) coupled to a microreactor is an excellent tool for the investigation of reactions in solution. Here, we report the first results of our investigations into preparatively interesting electron transfer-initiated chain reactions in solution which proceed via radical cations as reactive intermediates. The tris(p-bromophenyl)aminium hexachloroantimonate (1)-mediated [2+2] cycloaddition of trans-anethole (2) to give 1,2-bis(4 methoxyphenyl)-3,4-dimethylcyclobutane (3) was investigated. The reaction proceeds as a radical cation chain reaction via transient intermediates 2*+ and 3*+ that could be detected and characterized unambiguously directly in the reacting solution by ESI-MS/MS. The identity of the intermediates was confirmed by comparison with authentic MS/MS spectra of 2*+ and 3*+ obtained by atmospheric pressure chemical ionization mass spectrometry (APCI-MS). In addition, substrate and product can be monitored easily in the reacting solution by APCI-MS. PMID- 14551666 TI - Performance evaluation of two chelating ion-exchange sorbents for the fractionation of labile and inert metal species from aquatic media. AB - The performance of two iminodiacetate chelating resins, applied in an element fractionation scheme, was investigated for the characterization of marine and stream water samples collected in potentially contaminated harbor and industrial zones. The comparison involved an evaluation of the sorption/desorption behavior of Cd, Cu, Pb and Zn on Chelex-100, a sorbent extensively employed for metal speciation studies, and on Lewatit TP-207, a material with similar chelating characteristics for which no applications for fractionation studies are reported. A characterization of the latter resin in terms of the influence of sample pH on the uptake of metals from aqueous solutions and their subsequent acid elution was carried out. Fractionation studies were performed on natural water samples and model solutions resembling their composition, spiked with micromolar concentrations of the metals. The operationally defined fractionation scheme is based on dynamic adsorption of the resin-labile fractions of metals on microcolumns made of the sorbents. This stage is followed by a batch procedure in which the eluent from the column is contacted with fresh resin to discriminate between the relatively stable species unable to react with the resin during the column interaction time, and the strongly complexed metals that do not interact with the sorbent even for periods of several hours. Results were obtained from coarsely filtered sub-samples and from specimens passed through membrane filters of pore size 0.45 and 0.22 microm. The method was also assessed with model solutions of the metals containing variable concentrations (in the 10(-3)-10(-7) molar range) of complex forming agents such as nitrilotriacetic acid and trans 1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid. An additional batch method, in which a fixed amount of resin was contacted with sample solution for increasing periods of time and the amount of metal remaining in solution measured as a function of the elapsed time, was used to obtain information on the kinetics of dissociation of species formed in the presence of added complexants. The behavior of both resins was similar, demonstrating that the discrimination of species derived from experiments with Chelex-100 may be properly validated by independent fractionation tests carried out with Lewatit TP-207. The behavior of the sorbents is also discussed in terms of the possible influence of particulate or colloidal materials and of kinetic effects related to the presence of complexing agents. PMID- 14551667 TI - Gas chromatography-mass spectrometry with solid-phase microextraction method for determination of methyl salicylate and other volatile compounds in leaves of Lycopersicon esculentum. AB - Methyl salicylate (MeSA) in many plants is a important signaling compound, which plays an important role in a pathogen-induced defense response. In this paper, gas chromatography-mass spectrometry (GC-MS) with headspace solid-phase microextraction (HS-SPME) was developed for determination of MeSA and other volatile compounds in leaves of a tomato plant (Lycopersicon esculentum). Tomato leaves were ground under liquid nitrogen and sampled by HS-SPME, with a 100 microm polydimethylsiloxane fiber, and finally analyzed by GC-MS. Eighteen compounds in the leaves of tomato plant infested by tobacco mosaic virus (TMV) were separated and identified, among them MeSA, which was quantitatively analyzed by the standard addition method. MeSA concentrations higher than 2.0 microg g(-1) fresh weight accumulated in leaves of TMV-infested tomato plant as the defense response to TMV. A similar concentration of MeSA in the leaves of MeSA-treated tomato plant was also found. No MeSA in leaves of control tomato plant was detected. These findings suggest that MeSA might be a signaling compound in the tomato plant response to TMV. The present method for determination of MeSA required only simple sample preparation and no organic solvent, and provided an excellent relative standard deviation of less than 5.0% and a low detection limit of 10 ng g(-1) fresh weight for MeSA. These results show that GC-MS-HS-SPME is a simple, rapid and sensitive method for determination of MeSA and other plant signaling compounds in plant tissues. PMID- 14551668 TI - Development of quantitative vitellogenin-ELISAs for fish test species used in endocrine disruptor screening. AB - The yolk protein precursor vitellogenin (Vtg) in plasma has proved to be a simple and sensitive biomarker for assessing exposure of fish to environmental estrogens. Within international bodies such as the Organization for Economic Cooperation and Development (OECD) work is ongoing to develop screening and testing programmes for endocrine disrupting effects of new chemicals, and in the focus of this development are the fish test species common carp (Cyprinus carpio), fathead minnow (Pimephales promelas), zebrafish (Danio rerio) and Japanese medaka (Oryzias latipes). In this study we have developed quantitative enzyme linked immunosorbent assays (ELISAs) for Vtg in common carp/fathead minnow, zebrafish and Japanese medaka. The assays were developed using a combination of monoclonal and polyclonal fish Vtg antibodies in a sandwich format, using stabilized Vtg from the test species as a standard. The carp Vtg ELISA has a working range of 1-63 ng/mL, a minimal detection limit of 0.6 ng/mL, and may also be used for quantification of Vtg in fathead minnow. In fathead minnow whole-body homogenate samples, the practical detection limit is 400 ng/mL due to the matrix effect. The zebrafish Vtg ELISA has a working range of 0.5-63 ng/mL, a minimal detection limit of 0.4 ng/mL, and a practical detection limit of 200 ng/mL in whole-body homogenate samples. The medaka Vtg ELISA has a working range of 0.25-16 ng/mL, a minimal detection limit of 0.1 ng/mL, and a practical detection limit of 125 ng/mL in whole-body homogenate samples. The intra- and inter-assay variations were below 20% for all assays. The assays were evaluated with sets of representative samples spanning the wide dynamic range of Vtg-levels found in fish exposed to environmental estrogens, and all three assays are currently undergoing international inter-laboratory validation. PMID- 14551669 TI - A comparative study of mid-infrared diffuse reflection (DR) and attenuated total reflection (ATR) spectroscopy for the detection of fungal infection on RWA2-corn. AB - An investigation into the rapid detection of mycotoxin-producing fungi on corn by two mid-infrared spectroscopic techniques was undertaken. Corn samples from a single genotype (RWA2, blanks, and contaminated with Fusarium graminearum) were ground, sieved and, after appropriate sample preparation, subjected to mid infrared spectroscopy using two different accessories (diffuse reflection and attenuated total reflection). The measured spectra were evaluated with principal component analysis (PCA) and the blank and contaminated samples were classified by cluster analysis. Reference data for fungal metabolites were obtained with conventional methods. After extraction and clean-up, each sample was analyzed for the toxin deoxynivalenol (DON) by gas chromatography with electron capture detection (GC-ECD) and ergosterol (a parameter for the total fungal biomass) by high-performance liquid chromatography with diode array detection (HPLC-DAD). The concentration ranges for contaminated samples were 880-3600 microg/kg for ergosterol and 300-2600 microg/kg for DON. Classification efficiency was 100% for ATR spectra. DR spectra did not show as obvious a clustering of contaminated and blank samples. Results and trends were also observed in single spectra plots. Quantification using a PLS1 regression algorithm showed good correlation with DON reference data, but a rather high standard error of prediction (SEP) with 600 microg/kg (DR) and 490 microg/kg (ATR), respectively, for ergosterol. Comparing measurement procedures and results showed advantages for the ATR technique, mainly owing to its ease of use and the easier interpretation of results that were better with respect to classification and quantification. PMID- 14551671 TI - Serum ghrelin concentrations in patients receiving olanzapine or risperidone. AB - RATIONALE: Although enhanced appetite and weight gain are potential side effects of treatment with antipsychotic agents, particularly olanzapine and clozapine, the mechanism is poorly understood. OBJECTIVES: To test the hypothesis that ghrelin, a gastrointestinal hormone that enhances appetite, is involved in increased food intake and weight gain during treatment with antipsychotics. METHODS: Serum ghrelin concentrations were investigated in schizophrenic patients receiving olanzapine or risperidone, and in healthy volunteers. RESULTS: Serum ghrelin concentrations did not increase, but rather decreased, in patients treated with olanzapine or risperidone in comparison with healthy volunteers. No significant difference was found in serum ghrelin concentration between patients treated with olanzapine and risperidone. CONCLUSIONS: Our results indicate that ghrelin is not a direct cause of increased food intake and weight gain during treatment with olanzapine or risperidone, whereas ghrelin is associated with metabolic change in patients receiving these agents. PMID- 14551672 TI - Rat poison and food security in the People's Republic of China: focus on tetramethylene disulfotetramine (tetramine). AB - The last several years have seen a large number of mass poisonings in mainland China, particularly those caused by illicit rodenticides. One rat poison, tetramine (tetramethylene disulfotetramine) is responsible for a great percentage of death and injury in the People's Republic of China (PRC). Tetramine is an acutely toxic substance with human oral toxicity estimated as low as 0.1 mg/kg, and is widely available in open markets in mainland China--this despite being prohibited for manufacture or sale in the PRC. Being a GABA antagonist, and having an extremely potent effect on the brain stem, many victims can quickly fall into convulsions and die within hours following ingestion. With no known effective antidote at this time, clinical data from the PRC show that acute cases of tetramine poisoning are extremely difficult to treat. The widespread use of tetramine--including its reported sale at a Malaysian outdoor market in September 2002--could exacerbate its hazard to public health, particularly in those areas having large overseas Chinese populations. PMID- 14551673 TI - Cellular and molecular studies on cisplatin-induced apoptotic cell death in rat kidney. AB - Using morphological and molecular approaches, we characterized cisplatin-induced cell necrosis and apoptosis in rat kidney. Male Sprague-Dawley rats ( n=5 per group) received a single intraperitoneal injection of either cisplatin (5 mg/kg) or saline, and were killed on day 5. Functionally, cisplatin-treated rats developed polyuric acute renal failure. Morphologically, kidneys of cisplatin treated rats showed overt tubular necrosis associated with apoptosis in the corticomedullary junction. Cell necrosis was segment-specific and was distributed in radial fashion at the corticomedullary junction. The apoptosis was limited to discrete cells in apparently intact tubules in the vicinity of the necrosed tubules. The apoptotic changes were confirmed by TUNEL (TdT-mediated deoxyuridine triphosphate nick-end labeling) and staining for cleaved caspase-3. Analysis of outer medullary tissue for apoptosis-related molecules by RNase protection assay revealed a significant increase in the expression of pro-apoptotic mRNAs (caspases 1, 2, and 8, and Bax) in cisplatin-treated rats. On the other hand, the expression of mRNA for the anti-apoptotic Bcl-2 did not change, resulting in a decrease in relative ratio of Bcl-2/Bax, and thus favoring apoptosis. The above changes were paralleled by a marked increase in caspase-3 precursor, the executioner protease. Furthermore, these pro-apoptotic molecular changes were associated with a 3-fold increase in the activity of JNK1 in the outer medulla, but not in the cortex, of cisplatin-treated rat kidneys, localizing to the site of maximal apoptosis. Upregulation of JNK1 activity in the outer medulla was not accompanied by changes in the activities of ERK or p38 kinase. In conclusion, these data suggest that cisplatin-induced apoptotic cell death in native kidney may be mediated by cooperative activation of the JNK1 pathway and Bax in the outer medulla. PMID- 14551670 TI - Pharmacogenetic studies of alcohol self-administration and withdrawal. PMID- 14551674 TI - PCR-based identification of microcystin-producing genotypes of different cyanobacterial genera. AB - Microcystins are harmful hepatotoxins produced by many, but not all strains of the cyanobacterial genera Anabaena, Microcystis, Anabaena, Planktothrix, and Nostoc. Waterbodies have to be monitored for the mass development of toxic cyanobacteria; however, because of the close genetic relationship of microcystin producing and non-producing strains within a genus, identification of microcystin producers by morphological criteria is not possible. The genomes of microcystin producing cells contain mcy genes coding for the microcystin synthetase complex. Based on the sequence information of mcy genes from Microcystis and Planktothrix, a primer pair for PCR amplification of a mcyA gene fragment was designed. PCR with this primer pair is a powerful means to identify microcystin-producing strains of the genera Anabaena, Microcystis, and Planktothrix. Moreover, subsequent RFLP analysis of the PCR products generated genus-specific fragments and allowed the genus of the toxin producer to be identified. The assay can be used with DNA from field samples. PMID- 14551675 TI - Prevalence of vertebral fractures among patients with chronic obstructive pulmonary disease in Canada. AB - PURPOSE: Patients with chronic obstructive pulmonary disease (COPD) may be at higher risk for osteoporosis. The primary objective of this case-control study was to determine the prevalence of vertebral fractures among patients with COPD admitted to acute care compared with a gender- and age-matched control group. SUBJECTS AND METHODS: Subjects were identified by chart reviews from an acute care hospital in Hamilton, Ontario, in 1999, including patients who were over 50 years old. In total, 127 patients with ICD-9 codes specifying COPD were randomly selected and compared with 127 gender- and age-matched controls. Chest radiographs were interpreted by two radiologists who defined and graded vertebral fractures using Genant's method. Medications taken, or prescribed at discharge, were recorded from charts. RESULTS: The overall prevalence of at least one vertebral fracture was found to be 34/127 (26.8%) in the COPD patients compared with 30/127 (23.6%) in the controls ( p=0.556). A significantly greater proportion of COPD patients had at least one severe vertebral fracture (OR=3.75, 95% CI 1.24 to 11.3). Review of hospital chest X-ray reports indicated that only 12 of 64 (18.8%) patients with vertebral fractures identified by the study radiologists actually had a vertebral fracture noted in the report. The proportion of COPD patients with vertebral fractures who were discharged on osteoporosis therapy was 5/27 (18.5%). There was a suggestion of lower lung function, as measured by forced vital capacity (FVC%), in patients with severe vertebral fractures ( p=0.067). CONCLUSIONS: These data indicate that: (1) There is an increased proportion of COPD patients with severe vertebral fracture, and (2) Documentation and treatment of osteoporosis in acute care COPD patients is low. Therefore, there is a need to target this high-risk group for osteoporosis screening and potential clinical management. PMID- 14551676 TI - Differential effects of modern immunosuppressive agents on the development of intimal hyperplasia. AB - Modern immunosuppressive agents such as tacrolimus and rapamycin are claimed to be associated with a reduction in vascular narrowing, a central feature of chronic rejection. This study assesses the effect of cyclosporine, tacrolimus and rapamycin on the development of intimal thickening, fibrosis-associated genes and deposition of extracellular matrix (ECM) proteins in a model of intimal hyperplasia. Male Sprague-Dawley rats received either no treatment or 5 mg/kg cyclosporine, 0.1 mg/kg tacrolimus or 0.05 mg/kg rapamycin. Animals underwent left common carotid balloon angioplasty, and intima medial ratios, pro-fibrotic gene expression and ECM accumulation were calculated at 14 and 28 days. Cyclosporine was associated with increased intimal thickening compared to controls ( P < 0.004). Tacrolimus had no effect on intimal thickening, whilst rapamycin significantly inhibited intimal thickening at both 14 and 28 days ( P < 0.004 and P < 0.026, respectively). All groups significantly inhibited matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitor of metalloproteinases (TIMP) 1, transforming growth factor (TGF)-beta and collagen III expression at 14 days ( P < 0.001), but increased ECM deposition. However, rapamycin marginally reduced ECM deposition compared to cyclosporine ( P < 0.06). Treatment with cyclosporine was associated with worsening of vascular narrowing, whilst rapamycin showed a beneficial reduction in intimal thickening. Treatment with all immunosuppressive agents resulted in increased ECM deposition. Rapamycin may halt the progression of vascular narrowing compared to both cyclosporine and tacrolimus. PMID- 14551677 TI - Is cholangiography required for ex situ splitting of cadaveric livers? AB - Anomalous biliary anatomy is encountered frequently and, if not considered at the time of ex situ splitting of a cadaveric liver into left lateral segment and right lobe grafts, right-sided second-order ducts that enter the left system, or the segment IV duct, may be damaged, leading to biliary complications in the recipients. Bench cholangiography facilitates delineation of these anomalies, but if one considers the commonly encountered variations in biliary anatomy, in relation to the correct plane of division of the left hepatic duct (away from the hilum, close to the umbilical fissure), it is possible to avoid inadvertent injury to right-sided sectoral ducts. This approach, combined with careful probing of the ducts and absolute identification of the segment IV duct negates the contribution of cholangiography. PMID- 14551678 TI - Correlation between the Banff 97 classification of renal allograft biopsies and clinical outcome. AB - The 1997 fourth Banff meeting revised the consensus for describing transplant biopsies. We have conducted a retrospective analysis of biopsies correlated between the Banff 97 classification and clinical outcome. The patients ( n=149), who had a total of 404 biopsy-proven rejections, were assessed and the biopsies taken from these patients were re-examined and classified according to the Banff 97 classification. Morphological changes in the glomeruli (g), interstitium (i), tubules(t), and arterial vessels (v) were scored. Severity of acute rejection was statistically associated with unresponsiveness to anti-rejection treatment ( P<0.0001) and predicted an increased risk of graft failure ( P<0.05). Each quantitative criterion (g, i, t, and v) was also statistically associated with unresponsiveness to anti-rejection treatment. Mean serum creatinine levels were significantly higher in the groups graded Banff 97 type I-III after 1 and 2 years of follow-up. The Banff 97 classification correlated with reversibility of rejection episodes and long-term graft survival. PMID- 14551679 TI - Hyperreninemic hypoaldosteronism: a possible etiological factor of septic shock induced acute renal failure. AB - OBJECTIVE: Hyperreninemic hypoaldosteronism has been described in critically ill patients. The present study investigated the plasma aldosterone concentration (PAC) in septic shock patients and its relationship with clinical course. DESIGN AND SETTING: Prospective descriptive study in a medical intensive care unit (ICU) of a university hospital. PATIENTS: Forty-six consecutive patients with septic shock as defined by the ACCP/SCCM criteria. INTERVENTION: A corticotropin stimulation test, followed by treatment with low doses of hydrocortisone and fludrocortisone. MEASUREMENTS AND RESULTS: Plasma renin activity, PAC, and cortisol levels were measured before and after the test. PAC measurements were repeated for 1 week. Relevant clinical and laboratory variables were recorded for ICU stay. Patients were divided into two groups according to PAC/renin activity ratio: above 2 (n=24 patients) and below 2 n=22). Patients with PAC/renin activity less than 2 had higher total volume of infused fluid, serum creatinine level, and fractional excretion of sodium values; aldosterone and serum creatinine were negatively correlated. Hypoaldosteronism was reversible within 1 week. Duration of ICU stay (p=0.0026) and the need for renal replacement therapy (p=0.0021) were greater in the group with PAC/renin less than 2. CONCLUSIONS: Transient hyperreninemic hypoaldosteronism is common in patients with septic shock. These abnormal aldosterone levels are associated with greater sodium and fluid depletion and are followed by enhanced incidence of acute renal failure requiring renal replacement therapy and prolonged length of stay in ICU. PMID- 14551681 TI - "Pond poop" from propofol. PMID- 14551680 TI - Early antibiotic treatment (prophylaxis) of septic complications in severe acute necrotizing pancreatitis: a prospective, randomized, multicenter study comparing two regimens with imipenem-cilastatin. AB - OBJECTIVE: We compared two imipenem regimens for prevention of septic complications in patients with severe acute necrotizing pancreatitis (ANP). DESIGN AND SETTING: Prospective, randomized open clinical trial involving intensive care units of 14 Spanish Hospitals. PARTICIPANTS: 92 patients with ANP. INTERVENTIONS: Imipenem/cilastatin was administered at 500 mg four times daily starting at the time of diagnosis of ANP, within the first 96 h from the onset of symptoms. Patients were randomized to receive antibiotic prophylaxis either for 14 days (group 1) or at least for 14 days and as long as major systemic complications of the disease persisted (group 2). RESULTS: Antibiotic was maintained in group 2 for 19.7+/-10.9 days. The incidence of infected pancreatic necrosis, pancreatic abscess, and extrapancreatic infections was 11%, 17%, and 28% in group 1 and 17.4%, 13%, and 35% in group 2 (n.s.). Pancreatic or extrapancreatic infection by Candida albicans occurred in 7% and 22% of patients. Global mortality was 18.5% (10.9% secondary to septic complications), without differences between groups. In patients with persisting systemic complications at day 14 mortality was almost always secondary to septic complications and decreased from 25% (group 1) to 8.8% (group 2) by maintaining antibiotic prophylaxis. CONCLUSIONS: Compared to a 14-day imipenem prophylaxis, a longer antibiotic administration in patients with ANP is not associated with a reduction in the incidence of septic complications of the disease. However, prolonged imipenem administration in patients with persisting systemic complications tends to reduce mortality in ANP compared to a 14-days regimen. PMID- 14551682 TI - Maximum FIO2 in minimum time depending on the kind of resuscitation bag and oxygen flow. AB - OBJECTIVE: To analyze what FIO2 can be reached, and how long it takes using the different autoinflated resuscitation bags and increasing oxygen flows. DESIGN: Experimental analysis on the effect of three different models of autoinflated resuscitation bag and increasing oxygen flows in the final FIO2, and time spent to reach it. SETTING: Laboratory, with a gas analyzer and a lung simulator to measure inspired FIO2. INTERVENTIONS: Simulated cardiopulmonary resuscitation. Three different autoinflated resuscitation bags were studied; A, the classic one with oxygen delivery directly into the bag, without reservoir, B, a new one without the reservoir device; and C, a new one with the reservoir device properly implemented. Increasing oxygen flows were administered until FIO2 stabilized. RESULTS: With model A the maximum FIO2 reached was 0.73 in 70 s using a 20 l/min oxygen flow. With model B the maximum FIO2 reached was 0.65 in 90 s using a 20 l/min oxygen flow. The best FIO2 (0.99) was reached using model C in 55 s with 12 l/min oxygen flow. In the three models a high correlation between oxygen flow and FIO2 was found (r>0.8). CONCLUSIONS: It is mandatory to use model C resuscitation autoinflated bag with 12 l/min of oxygen flow during the resuscitation maneuvers. Using another autoinflated bag model, maximum oxygen flows (i.e., 20 l/min) are needed. The resuscitation autoinflated bags showed less effectiveness when they were not properly assembled. PMID- 14551683 TI - Comment on "Death by parenteral nutrition" by Marik and Pinsky. PMID- 14551685 TI - A novel locus for clubroot resistance in Brassica rapa and its linkage markers. AB - An inbred turnip ( Brassica rapa syn. campestris) line, N-WMR-3, which carries the trait of clubroot resistance (CR) from a European turnip, Milan White, was crossed with a clubroot-susceptible doubled haploid line, A9709. A segregating F(3) population was obtained by single-seed descent of F(2) plants and used for a genetic analysis. Segregation of CR in the F(3) population suggested that CR is controlled by a major gene. Two RAPD markers, OPC11-1 and OPC11-2, were obtained as candidates of linkage markers by bulked segregant analysis. These were converted to sequence-tagged site markers, by cloning and sequencing of the polymorphic bands, and named OPC11-1S and OPC11-2S, respectively. The specific primer pairs for OPC11-1S amplified a clear dominant band, while the primer pairs for OPC11-2S resulted in co-dominant bands. Frequency distributions and statistical analyses indicate the presence of a major dominant CR gene linked to these two markers. The present marker for CR was independent of the previously found CR loci, Crr1 and Crr2. Genotypic distribution and statistical analyses did not show any evidence of CR alleles on Crr1 and Crr2 loci in N-WMR-3. The present study clearly demonstrates that B. rapa has at least three CR loci. Therefore, the new CR locus was named Crr3. The present locus may be useful in breeding CR Chinese cabbage cultivars to overcome the decay of present CR cultivars. PMID- 14551684 TI - Globular adiponectin upregulates nitric oxide production in vascular endothelial cells. AB - AIMS/HYPOTHESIS: Adiponectin, also called ACRP30, is a novel adipose tissue specific protein that has been shown to improve insulin sensitivity and to exert anti-atherogenic effects. It is known that knockout mice lacking endothelial NO synthase (eNOS) develop hypertension, insulin resistance, hyperlipidaemia, and show augmented ischaemia-reperfusion damage. Thus, we examined whether globular adiponectin activates eNOS to produce NO. METHODS: To analyze NO production in bovine aortic endothelial cells (BAE), NOx (nitrite and nitrate) was measured in the medium with an automated NO detector/high-performance liquid chromatography system. eNOS activation was assessed by phosphorylation of the enzyme and its activity was evaluated by citrulline synthesis in human umbilical vein endothelial cells (HUVEC). eNOS mRNA and protein expressions in HUVEC were evaluated by Realtime PCR and Western blot analysis. RESULTS: Gobular adiponectin increased NO production in BAE. It also caused eNOS phosphorylation and potentiated eNOS activity in HUVEC. In addition, globular adiponectin up regulated the eNOS gene to increase protein expression in HUVEC. CONCLUSION/INTERPRETATION: Globular adiponectin increases NO production through two mechanisms, namely, by activation of eNOS enzyme activity and via an increase in eNOS expression. Activation and up-regulation of eNOS could explain some of the observed vasoprotective properties of globular adiponectin, as well as its beneficial effects on the cardiovascular system. PMID- 14551686 TI - [Lamotrigine in women with epilepsy. Review of present data]. AB - The manuscript deals with the tolerability of Lamotrigine in women. The recent literature is reviewed with respect to interactions with oral contraceptives, sexuality, infertility, interactions with sex hormones, polycystic ovarian syndrome, adipositas, cosmetic side effects, osteoporosis, pregnancy, breast feeding, and teratogenetic effects. The available data have practical implications for the safe use of Lamotrigine in women. PMID- 14551687 TI - [Evidence-based arm rehabilitation--a systematic review of the literature]. AB - Based on a systematic MEDLINE search and informal sources, 40 references were identified that evaluate training therapy or neuromuscular electric stimulation for arm paresis after stroke and describe either a systematic review, meta analysis, randomised controlled trial, or controlled cohort study. The evidence was grouped into three areas of interest: comparison of physiotherapy schools, effects of intensity of training, and efficacy of specific arm rehabilitation techniques. The only physiotherapy school with evidence of superior efficacy was the task-oriented 'motor relearning programme'. Higher intensities of motor rehabilitation can accelerate motor recovery. Various training techniques with demonstrated efficacy are available for specific patient subgroups: arm ability training for mildly affected patients with reduced efficiency of motor control, constrained-induced movement therapy for patients with partial functional deficits and learned nonuse of the affected arm, and repetitive sensorimotor training techniques, EMG-biofeedback, functional electrical stimulation, and robot-assisted training for patients with severe arm paresis. PMID- 14551688 TI - [Chemokine--possible new options for the treatment of multiple sclerosis]. AB - Accumulation and activation of mononuclear cells (lymphocytes and monocytes) in the CNS is one of the crucial steps in the pathogenesis of multiple sclerosis (MS). Chemokines and their receptors govern physiological and pathological leukocyte trafficking and may also be pertinent in hematogenous leukocyte infiltration of the CNS. Due to broad pharmacological interest in the chemokine system, peptide antagonists and small molecular antagonists are now available for clinical therapeutic trials. For the treatment of MS in particular, the chemokine receptors CCR1, CCR2, CCR5, and CXCR3 are possible targets in a chemokine-based therapeutic approach. In this review, we summarize current knowledge of the roles of chemokines and chemokine receptors in the pathogenesis of MS. Furthermore, options for possible therapeutic intervention through the chemokine system are outlined. Clinical studies in MS patients applying this knowledge are expected soon. PMID- 14551689 TI - [The human vomeronasal organ]. AB - Odors influence human behavior. The perception of so-called pheromones is frequently mentioned in the context of a functional vomeronasal organ. Vomeronasal ducts can be detected in approximately half of the population. Its functionality, still a matter of debate, seems to be unlikely, at least after birth. It is easily conceivable that pheromone-induced changes in behavior are mediated through receptors in the human olfactory epithelium. PMID- 14551690 TI - [Visual search in healthy persons and Alzheimer's patients: relating cognitive function to clinical practice]. AB - Visual search is a complex and highly relevant cognitive task. Hypotheses about the processes involved have been derived from experimental psychology and modified by recent functional imaging methods revealing the underlying neuronal networks. To close the gap between theoretical implications and clinical practice, models of visual search are employed to explain visual search impairments in patients with Alzheimer's disease. PMID- 14551691 TI - [Altered cerebral excitability and spreading depression. Causes for the comorbidity of epilepsy and migraine?]. AB - Increased co-occurrence and common clinical aspects of epilepsy and migraine lead to the question of a common pathophysiological model of the diseases. Shared genetic risk factors as an explanation for comorbidity could not be proven. Clinical studies underline the unspecific association of migraine and epilepsy. Comorbidity is based on spreading depression as an expression of altered brain state with neuronal hyperexcitability. In comorbid conditions, therapy with valproate, gabapentin, or topiramate may be effective. PMID- 14551692 TI - [Psychosomatic liaison service in neurology. Accordance between need for psychotherapeutic interventions and their realization]. AB - The study addresses the realization of psychotherapeutic interventions in the framework of an integrated psychosomatic liaison service in neurology. Eighty four patients from one unit at the Department of Neurology of the University of Freiburg, Germany, underwent structured psychodiagnostic interviews and filled in self-rating instruments to evaluate mental disorders, psychosocial distress, ways of coping, and quality of life. Motivation and the need for treatment were estimated by both patients and a liaison psychotherapist. Using ICD-10-F criteria, mental and behavioral disorders were diagnosed in 35% of the cases. Thirty-seven percent of the patients desired psychological support. Actual intervention occurred in 27%, with a mean consultation rate of 3.7 sessions. There was only slight agreement between the estimated need for treatment and the interventions actually performed. The realization of psychotherapeutic interventions was subject to a complex interaction between patient needs, "objective" psychosocial distress, and institutional factors. Differentiation between primary medical care and psychotherapy was difficult. PMID- 14551693 TI - [Electrophysiological impairment profile of patients with Wilson's disease]. AB - In addition to hepatic and extrapyramidal motor clinical symptoms, Wilson's disease patients also exhibit subclinical disorders of other central nervous pathways. In this study, an impairment profile is described by means of eight electrophysiological tests (EAEP, MSEP, TSEP, T-VEP, MEP, EEG, heart frequency variability, and SSR) for 37 patients (28 with neurological, nine with tnon neurological form) undergoing long-term drug therapy. The occurrence in 64.3% of a delayed wave III and/or IPL III-V prolongation in patients with the neurological form makes pathological FAEP the most common form of the disorder, followed by disorders in MSEP, TSEP, MEP, and T-VEP. Patients with the non neurological form usually have normal values, although latency prolongations occur in isolated cases. The range of evoked potential findings is characterised primarily by latency prolongations, i.e. a demyelinising impairment type, and significant losses of potential hardly occur (except in the MEP). The electrophysiological impairment profile does not include EEG changes or vegetative disorders. PMID- 14551694 TI - [Polyneuropathy as a sole syndrome in malignant thymoma]. AB - Up to 40% of patients with malignant thymoma suffer from paraneoplastic symptoms (90% myasthenia, 10% other symptoms). A 55-year-old patient developed ascending symmetrical sensorimotor tetraparesis. A malignant thymoma without metastases was diagnosed 6 months later. Despite thymectomy followed by radiation and high-dose corticosteroid therapy, the polyneuropathy progressed. Six months after onset, the patient was bound to a wheelchair. Immunosuppressive therapy with cyclophosphamide was initiated, leading to marked remission. After ten cycles, the patient was able to walk independently with walking aids. After the sixth and tenth cycle, respectively, attempts to discontinue immunosuppression led to relapse. In several diagnostic workups, however, there was no tumour relapse. After 13 cycles, cyclophosphamide was replaced by immunoglobulins (0.4 g/kg per day i.v. for 5 days/month) due to progressive renal failure. The patient died just before the second course of this treatment. In conclusion, in the differential diagnosis of rapidly progressive polyneuropathy, a malignant thymoma should be considered, even in the absence of myasthenia. Immunosuppression with cyclophosphamide resulted in amelioration of symptoms in this patient. PMID- 14551695 TI - [Cerebral contrast medium extravasation after coronary angioplasty]. AB - We report on an 82-year-old woman who suffered a sudden loss of consciousness, right-sided hemiparesis, and aphasia during a coronary angiographic procedure. Computed tomography (CT) of the brain performed immediately revealed an edema and extravascularly localized contrast media in the left hemisphere. Within 6 h, neurological symptoms had disappeared, and a second CT after 48 h revealed normal results. Usually, extravasation of contrast media affects the posterior circulation with cortical blindness. This case demonstrates that contrast media may affect the blood-brain barrier also outside the posterior circulation. If neurological symptoms occur during angiography, contrast media extravasation must be distinguished from embolism or vasospasm induced by the angiographic procedure. PMID- 14551696 TI - [Diagnosis and differential diagnosis of lysosomal glycogen storage disease]. AB - We report a 73-year-old patient with acid maltase deficiency who initially had been suspected of having motor neuron disease. We discuss clinical and electrophysiological features of muscular lysosomal glycogenosis, with special emphasis on the histopathological differential diagnosis of the disease. PMID- 14551697 TI - [Niemann-Pick disease type C--a neurometabolic disease through disturbed intracellular lipid transport]. AB - Niemann-Pick disease type C (NPC) is a rare, neurovisceral lipid storage disorder caused by genetic defects in lipid transporting proteins. It is distinct from Niemann-Pick types A and B (sphingomyelin lipidoses) and displays genetic (mutations in the NPC1 or NPC2[=HE1] gene), biochemical, and clinical heterogeneity. Late infantile to juvenile forms of NPC predominate and are characterised by atypical behaviour, ataxia, dysarthria, dysphagia, dystonia, cataplexy, vertical gaze palsy, splenomegaly, and dementia. In adult variants, psychosis and dementia are common, and dysarthria, ataxia, splenomegaly, and vertical gaze palsy are further facultative signs. Routine laboratory results including serum cholesterol are normal. In bone marrow smears, sea-blue histiocytes are often demonstrated and foam cells sometimes seen. The diagnosis is confirmed by detecting free cholesterol accumulation in perinuclear granules (lysosomes) and reduced cholesterol esterification after challenge with exogenous low-density lipoprotein in fibroblasts. Alternatively or additionally, mutational analysis can be performed. Treatment is restricted to symptomatic measures, since there is no specific therapy. PMID- 14551698 TI - [Mitoxanthrone in the therapy of multiple sclerosis]. AB - The recent publication of the MIMS study (Mitoxantrone in multiple sclerosis), approval by local health authorities, and the recommendation by the MS Therapy Consensus Group (MSTCG) support the use of mitoxantrone in patients with active multiple sclerosis. This review provides an outline of relevant preclinical and clinical studies, discusses relevant side effects of the compound, and compares its use with other therapeutic approaches to this crippling disorder. PMID- 14551699 TI - [The most common form of dizziness in middle age: phobic postural vertigo]. AB - Up to now, there have been only a few valid epidemiological investigations of dizziness or vertigo as key symptoms. According to an analysis of 4,214 patients examined between 1989 and 2002 in an outpatient dizziness unit, benign peripheral paroxysmal positional vertigo (BPPV) and phobic postural vertigo (PPV) constitute the two most frequent syndromes. In this study, the relative age and sex distribution of both disorders was analysed. In the age group from 20 to 50 years, PPV was comparatively the most frequent form of dizziness, with a share of 22% to 26%. When left untreated, PPV becomes chronic in most cases and leads to considerable impairments, also at work. However, when diagnosed correctly, it can be treated successfully in more than 70% of cases. Thus, it takes on considerable medical and socioeconomic significance and should be part of the diagnostic repertoire of every doctor. PMID- 14551700 TI - [Vladimir Kernig (1840-1917)]. PMID- 14551701 TI - Paraoxonase gene polymorphisms, oxidative stress, and diseases. AB - The paraoxonase (PON) gene cluster contains at least three members, including PON1, PON2, and PON3, located on chromosome 7q21.3-22.1. Until now there has been little insight into the role of the respective gene products in human physiology and pathology. However, emerging evidence from biochemical and genetic experiments is providing clues about the role(s) of the products of these genes, which indicates that PON(s) acts as important guardians against cellular damage from toxic agents, such as organophosphates, oxidized lipids in the plasma low density lipoproteins. In parallel, substantial data have been published on the association between the polymorphisms of PON(s) and coronary heart disease. It has become clear that the polymorphisms significantly affect the prevalence of coronary heart disease. However, the associations between the PON(s) polymorphisms and most of these conditions were found to be inconsistent when additional populations were investigated. This contribution provides an overview of the status of research of each of the three genes and the available association studies and the potential problems in interpreting the data. We also review the current evidence on the association between PON(s) polymorphisms and diseases other than coronary heart disease and some metabolic quantitative phenotypes, such as plasma lipoproteins, plasma glucose, and birthweight. Finally, we suggest directions for the future that might elucidate the role of the PON genetic polymorphisms in this potentially important function of PON(s) and the role in coronary heart disease and other related diseases. PMID- 14551703 TI - A novel, alternative pathway of apoptosis triggered through class II major histocompatibility complex molecules. AB - Major histocompatibility complex class II (MHC-II) molecules, in addition to their role of presenting antigen to T lymphocytes, can serve as receptors triggering programmed cell death. MHC-II induced cell death affects activated/tumour transformed cells selectively, and it proceeds without the involvement of caspases, the major proteases of classical apoptosis. Caspase independent programmed cell death can also be triggered, albeit less effectively, via a series of other cell surface molecules. Here, we discuss the major characteristics, physiological significance, and clinical relevance of caspase independent apoptotic pathways with particular emphasis on the one induced by MHC II ligation. PMID- 14551704 TI - [Can communication with terminally ill patients be taught? Evaluation of a course model]. AB - BACKGROUND AND PURPOSE: Communication with terminally ill patients is a main responsibility of physicians. However, many physicians feel insufficiently prepared for this task. Models of courses resulting in improvements of communicative skills of participants have been published mainly in the Anglo American literature. This study describes the realization of a 2-day course model based on the experiences of the first three courses of this kind in Rhineland Palatinate, and analyzes changes of participants' communication behavior. METHODS: After each seminary, an evaluation form concerning participants' satisfaction with the course was filled in. Furthermore, all course participants received a questionnaire at the beginning and at the end of the course, as well as 3 months afterwards. The participants were asked to assess their own sense of security in seven different communication settings on a visual analog scale, and to specify perceived changes in their communication behavior 3 months after the course. RESULTS: The first three courses were attended by 31 participants. Course evaluation revealed high satisfaction scores with methods as well as with clarity and relevance of the contents. Self-assessment of participants showed a growing sense of security in different communication settings. Important increases could be demonstrated for communicating a diagnosis of cancer with good or less good prognosis, recurrence of cancer or a far progressive cancer disease without curative approach. 3 months after the course, participants described multiple changes indicating increased sensibility and professionalism in communication behavior. CONCLUSION: The realized communication skills courses resulted in relevant changes in communication behaviour and self-confidence of participants. Communication with terminally ill patients can be taught. PMID- 14551702 TI - Cardiac mechanotransduction and implications for heart disease. AB - Mechanotransduction, the conversion of a mechanical stimulus into a cellular response, plays a fundamental role in cell volume regulation, fertilization, gravitaxis, proprioception, and the senses of hearing, touch, and balance. Mechanotransduction also fills important functions in the myocardium, where each cycle of contraction and relaxation leads to dynamic deformations. Since the initial observation of stretch induced muscle growth, our understanding of this complex field has been steadily growing, but remains incomplete. For example, the mechanism by which myocytes sense mechanical forces is still unknown. It is also unknown which mechanism converts such a stimulus into an electrochemical signal, and how this information is transferred to the nucleus. Is there a subpopulation of mechanosensing myocytes or mechanosensing cells in the myocardium? The following article offers an overview of the fundamental processes of mechanical stretch sensing in myocytes and recent advances in our understanding of this increasingly important field. Special emphasis is placed on the unique cardiac cytoskeletal structure and related Z-disc proteins. PMID- 14551706 TI - [Treatment of thrombosis with low-molecular-weight heparin. Comparison of body weight-adjusted and fixed dosage]. AB - BACKGROUND: The treatment of deep vein thrombosis (DVT) has been simplified and become safer after introduction of low-molecular-weight heparin (LMWH). The dosage of LMWH was performed by body weight adjustment without dose-finding studies. DATA: In three large clinical trials, the improved efficacy and safety of body weight-independent fixed-dose LMWH Certoparin were demonstrated using 2 x 8,000 IU subcutaneously per day for the initial treatment of symptomatic acute DVT. The reduction of thrombus size after 14 days, determined with the Marder score, and the combined endpoint, consisting of recurrent thromboembolism, major bleeding and mortality, was lower over the initial treatment with LMWH compared to intravenous APTT-adjusted unfractionated heparin (UFH). The benefit of the clinical endpoints over 6 months was demonstrated in two clinical trials. CONCLUSION: Treatment of acute DVT is more effective and safer using fixeddose 2 x 8,000 IU Certoparin for the initial treatment compared to UFH. PMID- 14551705 TI - [Optimized diabetes therapy in type 2 diabetics]. AB - BACKGROUND: With the prevalence of type 2 diabetes mellitus having increased to approximately 8% during recent years and a further rise likely, type 2 diabetes will develop into a major health care problem in Europe. There are roughly 8 million diabetics in Germany, with another 4 million estimated unknown case. CLINICAL FEATURES: Diabetes is an accepted risk factor in the development of micro- and macrovascular complications. These complications generate enormous costs in our health care system. The optimal treatment of diabetic complications will minimize the cost to the health care system in the long run. Therefore, it is necessary to relay current therapeutic guidelines to medical personnel on a widespread basis. AIMS AND CONCLUSION: This publication summarizes current therapeutic options in glycemic control of type 2 diabetics which correspond to 95% of all diabetics. Recently, a national guideline has been developed by the "Arzneimittelkommission der deutschen Arzteschaft", the "Deutsche Diabetes- Gesellschaft", the "Fachkommission Diabetes Sachsen", the "Deutsche Gesellschaft fur Innere Medizin e. V.", and the "Arbeitsgemeinschaft der Wissenschaftlichen Medizinischen Fachgesellschaften". In this guideline, treatment recommendations are made according to the criteria of evidence-based medicine. It is the basis of this review. In addition, the significance of the modern option of functional insulin therapy (FIT) in the treatment of type 2 diabetes will be explained. In closing, innovative alternatives of applying insulin will be discussed. Unquestionably, all other risk factors such as hypertension and/or hyperlipidemia also have to be treated optimally. These risk factors will not be discussed in this publication. PMID- 14551707 TI - [Diagnosis of cholestatic disorders]. AB - BACKGROUND: Cholestasis is known as an etiologically diverse clinical entity which requires a broad differential diagnostic workup. In the majority of patients, history, clinical examination, clinical chemical analysis, and abdominal ultrasound enable the differentiation between extrahepatic and intrahepatic cholestasis. This review summarizes our current knowledge in the diagnosis of cholestatic disorders. METHODS: In regard to clinical practice, diagnostic tools and new developments in imaging and molecular genetics are discussed including an algorithm for the diagnostic workup of cholestatic patients. CONCLUSION: Ultrasound and computed tomography have represented the most important primary imaging techniques in hepatobiliary disorders over the last 2 decades. The direct visualization either by endoscopic retrograde cholangiopancreatography (ERCP) or percutaneous transhepatic cholangiography (PTC) still remains the gold standard in the evaluation of the extrahepatic bile duct. In the past decade, magnetic resonance cholangiopancreatography (MRCP) has increasingly been established as a noninvasive alternative, thereby reducing the necessity of ERCP as an invasive exploration of the biliary system. Liver biopsy is indicated for the histologic grading and staging of intrahepatic cholestatic disorders. Recently, molecular genetic studies have elucidated several mutations in genes of hepatobiliary transporters which are responsible for hereditary forms of cholestasis in man. Thus, molecular genetics may be of interest in single cases of unclassified cholestasis or familial syndromes and will contribute to the routine diagnosis of hereditary cholestatic syndromes in the future. In summary, application of these diagnostic tools will finally lead to an unequivocal diagnosis in the majority of cholestatic patients with consecutive rational therapy. PMID- 14551708 TI - [Treatment of hypertension in the elderly]. PMID- 14551709 TI - [Rheumatology 2003-part I: research news concerning pathogenesis, epidemiology, diagnosis, and therapy of chronic inflammatory joint diseases]. AB - Due to the partial elucidation of the immunopathogenesis of chronic inflammatory diseases during the last years, clinical rheumatology has made a rapid development, which by the consequent use of immunomodulatory therapies including recombinant proteins (biologicals) led to a significantly ameliorated prognosis of these diseases. On this basis, new research projects are continuously performed in the fields of pathogenesis, new drug development, outcome and therapy studies. New developments of imaging techniques and serologic testing facilitate a better classification and definition of disease activity and remission criteria. The current state of research in the field of rheumatoid arthritis and spondylarthropathies with its clinical consequences is reviewed in this article on the basis of the most recent data available. PMID- 14551710 TI - [28-Year-Old Patient with Elevated Serum IgE Levels and Multiple Refractory Epidural and Paravertebral Abscesses. Clinical manifestation of a Job's Syndrome?]. AB - BACKGROUND: The hyper-IgE syndrome is a rare primary immunodeficiency characterized by recurrent staphylococcal infections and high levels of serum IgE. CASE REPORT: The case of a 28-year-old man with epidural and paravertebral abscesses and a markedly elevated serum IgE level of 2,609 U/ml is reported. There was no allergic diathesis, nor had the patient a history of other diseases, which might explain the high serum IgE level. In a blood culture, Staphylococcus aureus was detected. Since early childhood, the patient has suffered from recurrent skin infections. CONCLUSION: The presented study case discusses the diagnosis of a Job's syndrome in this patient and reviews previously published cases. It also summarizes the current knowledge about pathogenesis and diagnostic criteria of this rare syndrome. PMID- 14551711 TI - Using porcine small intestinal submucosa in intestinal regeneration. AB - Small intestinal submucosa (SIS) is an unusual tissue that promotes constructive tissue remodeling when applied as a xenogeneic material. The aim of our experimental study was to assess its effectiveness in intestinal regeneration. Twenty white New Zealand rabbits were anesthetized and underwent celiotomy. A 6 cm antimesenteric incision was created at the jejunal segment. An elliptical SIS graft measuring 6 cm long and 2 cm wide was sutured to the jejunal defect as a patch graft. Thirteen living rabbits were divided into groups of three and the grafts were harvested at postoperative weeks 2, 4, and 6. The obtained specimens were evaluated for gross and histologic appearance. In morphometric examination, in the 2, 4, and 6 weeks groups, the diameters of grafted intestines were larger than preoperatively by 50%, 25%, and 25% respectively; also the grafts had contracted to 0%, 25%, and 50% of their original sizes respectively. At the end of 2 weeks, the grafts were intact without evidence of epithelial regeneration. By 4 weeks, intestinal tissue regeneration was started, and epithelial coverage of the grafts was detected. The grafts were covered with a complete intestinal mucosa at 6 weeks. Remarkable regeneration marked fibroplasia, angiogenesis, and mild mononuclear cell infiltration had also occurred throughout the grafts at 6 weeks. Porcine SIS appeared an effective biodegradable scaffold, facilitating regeneration of intestinal tissue. These results suggest that SIS may be useful to increase the mucosal surface of intestine and may provide a new substance for short gut syndrome in the future. PMID- 14551712 TI - Treatment of the toe tourniquet syndrome in infants. AB - INTRODUCTION: The "toe tourniquet syndrome" is the circumferential strangulation by human hair or fibers of one or more toes in infants, which may induce prolonged ischemic injury and tissue necrosis. Release of the strangulation is mandatory to avoid autoamputation of the digit. We recently encountered several incompletely treated cases and would like to emphasize the effective method of treatment. METHODS: A retrospective analysis of pediatric emergency department records of patients treated for "toe tourniquet syndrome" during 1990 to 2001 was performed. Patients undergo a short, longitudinal, deep incision over the area of strangulation on the dorsal aspect of the toe, until the phalanx bone, which allows the complete section of the constricting fibers to be removed without injury to the anatomical structures of the toe. RESULTS: Twenty-one infants (12 boys and nine girls), aged 0-11 months (average 4.6 months), were treated for tourniquet syndrome of one (15 infants) or two (six patients) toes. In 12 patients (57.1%) the right foot was involved. Eight patients (38.1%) had undergone an attempt in the previous few days to release constriction. No complications were encountered. CONCLUSION: The above-described technique is simple and safe and secures the complete release of the strangulation by removal of all hairs or fibers without injury to the anatomical structures of the toe. PMID- 14551713 TI - Intralobar pulmonary sequestration with congenital cystic adematous malformation and rhabdomyomatous dysplasia. AB - Pulmonary sequestration and congenital cystic adenomatous malformations (CCAM) are well known but still uncommon anomalies of the lung. Extralobar sequestrations are well described in association with CCAM, but fewer intralobar lesions are found with this association. The existence of striated muscle within CCAM is described, and we have evidence of dysplastic changes within the various cellular components occurring, with rhabdomyomatous dysplasia being one of these. A literature review shows no previous evidence of a reported intralobar sequestration associated with CCAM and rhabdomyomatous dysplastic changes. PMID- 14551715 TI - Malrotation as a preparing ground for intussusception. PMID- 14551714 TI - Necrotizing amebic colitis: the plea for an early diagnosis. PMID- 14551716 TI - "Thoracic gastrostomy"--a new technique for feeding gastrostomy in wide-gap esophageal atresia and tracheo-esophageal fistula. AB - INTRODUCTION: Initial management of wide-gap esophageal atresia and tracheo esophageal fistula, especially in developing countries, often involves ligation of the fistula, proximal esophagostomy and a gastrostomy. The conventional gastrostomy requires an abdominal incision and has several complications. The authors present an alternative technique of 'gastrostomy' that does not require an abdominal incision. METHOD: Through a standard right thoracotomy, the fistula is divided and tracheal end closed. If primary anastomosis is not possible due to wide gap, the lower esophageal pouch is mobilised and brought out as a stoma on the left posterior chest wall, lateral to the paraspinal muscles in the 7th or 8th intercostal space. A feeding tube is passed through the stoma into the stomach. The thorax is closed with a drain. Proximal esophagostomy is performed in the neck. The thoracic stoma is intermittently intubated for feeding until the child is ready for gastric transposition. RESULTS: The technique was successfully used in 5 patients. Feeding through the stoma could be established in 4. One patient had stomal retraction and died of resultant sepsis. One patient has subsequently undergone gastric transposition and one more is waiting for it. Two patients died of unrelated causes before esophageal replacement could be carried out. DISCUSSION: This is an alternative technique for feeding 'gastrostomy'. We have named the procedure as "Thoracic Gastrostomy" because the stoma is located on the chest but functions as a gastrostomy. The advantages include avoidance of a laparotomy and its complications, easy feeding by intermittent intubation, and availability of a virgin stomach for future gastric transposition. PMID- 14551718 TI - Does high-pressure carbon dioxide insufflation facilitate mucosal dissection in transanal endorectal pull-through? A rabbit model. AB - The aim of the present study was to investigate whether high-pressure carbon dioxide insufflation facilitates mucosal dissection in the transanal endorectal pull-through (TEPT) operation performed in Hirschsprung's disease in a rabbit model. In the study, ten New Zealand rabbits were used. In six of the rabbits, CO(2) under 50 mmHg pressure was insufflated into the submucosal area through a 25-gauge scalp needle inserted 0.3 cm over the dentate line. Two of the rabbits were taken for histopathological examination. In four rabbits, the TEPT operation was performed and in another four the operation was performed without CO(2) insufflation. Histopathological examination in the CO(2)-insufflated group of rabbits showed that at transversal incisions mucosa was seen to be separated circumferentially from submucosa by high pressure CO(2) and at longitudinal incision the separation was seen to be complete along the anal canal. It was noted that mucosal dissection was rapid, non-bloody and without induced mucosal tears or perforation; however, in the non-insufflated group the operation was time-consuming, bloody and more difficult to perform and needed meticulous care. In the present study, it was seen that submucosal high pressure CO(2) insufflation in rabbits facilitates mucosal dissection in the TEPT operation and is easy to perform, time-saving and highly economical. PMID- 14551717 TI - Pathogenesis of neonatal necrotizing enterocolitis: a study of the role of intraluminal pressure, age and bacterial concentration. AB - The pathogenesis of neonatal necrotizing enterocolitis (NEC) is unknown. Intestinal dilatation and preferred occurrence of NEC at sites of bacterial overgrowth (colon and ileum) are common findings. The study attempted to produce NEC with increasing intraluminal pressures and bacterial concentrations in two different aged groups of rats. First, 10-cm terminal ileum segments were isolated with intact vascular pedicles in 1-and 3-month-old rats, and a dose of 10(11) E. coli in 1 ml was injected into each segment. Intraluminal pressure was sustained for 1 h at 150, 100, 50 and 0 cmH(2)0, respectively, in four experimental groups ( n=6). The isolated loop was then returned to the abdominal cavity and assessed grossly for NEC after 24 h. Histological examination was performed by a pathologist (KWC) who was blinded to the procedures. Second, the procedure was repeated with doses of 10(8), 10(5) and 0 bacteria/ml ( n=6) at intraluminal pressure of 100 cmH(2)0 in 1-month-old rats. Third, in another experimental group, oxygenation of the pedicled loop was assessed by oximetry as the intraluminal pressure increased and the findings were correlated with aortic blood pressure. The blood pressures (mean+/-SD) for 3- and 1-month-old rats were 110+/-6 and 72+/-4 mmHg, respectively. Hypoxia (<50% oxygen saturation) of the bowel was detected when the intraluminal pressure exceeded the mean blood pressure. The relative incidences of NEC in the bowel with intraluminal pressure above and below mean blood pressure were 100% (6/6) vs. 4% (1/24; P<0.05) in 3 month-old rats, and 100% (12/12) vs. 11% (2/18; P<0.05) in 1-month-old rats. There was no occurrence of NEC in bowel injected with 10(5) E. coli/ml and less at 100 cm intraluminal pressure. Increased intraluminal pressure results in bowel hypoxia and in the presence of adequate bacterial concentration predisposes to the development of NEC. Young age is associated with a lower threshold for increased intraluminal pressure leading to NEC. PMID- 14551719 TI - New long-acting androgens. AB - Testosterone substitution treatment aims to replace physiological actions of endogenous testosterone by steadily maintaining physiological blood levels of testosterone. The underlying conditions rendering androgen replacement necessary are usually irreversible. The consequence is that almost life-long androgen replacement is required. Patient compliance with life-long androgen replacement depends on convenient pharmaceutical formulations ensuring continuity of androgen replacement. Therefore, they must be convenient in usage with a relative independence of medical services. In elderly man, safety of androgen replacement therapy is a concern but in younger subjects (below the age of 50 years) side effects of androgens are usually minimal. For them, long-acting testosterone preparations are well suited. Testosterone implants generate, depending on the dose of implants, 3-6 months of normal plasma testosterone. This method requires minor surgery. Injectable testosterone undecanoate maintains plasma testosterone in the normal range for 12 weeks. PMID- 14551721 TI - Transdermal testosterone delivery: testosterone patch and gel. AB - Testosterone replacement treatment is usually life-long. Fortunately, testosterone administration is relatively safe and until the age of 50 years few side effects are noted with normal doses of testosterone. After the age of 50 years when prostate disease becomes more prevalent, shorter-acting testosterone preparations, allowing a fast reduction of circulating testosterone levels, may be an advantage. Testosterone has an impact on sexual and non-sexual behaviour and short-acting testosterone preparations may be better suited for the initiation of long-term administration allowing the monitoring of behavioural effects. Testosterone can be delivered to the circulation through the intact skin, both genital and non-genital. Transdermal administration delivers testosterone at a controlled rate into the systemic circulation, avoiding hepatic first pass and reproducing the diurnal rhythm of testosterone secretion and without the peak and trough levels observed with the use of the traditional long acting testosterone injections. In conclusion, both the testosterone patch and testosterone gel are valuable contributions to androgen replacement treatment meeting the requirements specified for testosterone replacement treatment. PMID- 14551720 TI - Androgen therapy with dehydroepiandrosterone. AB - The physiological role of dehydroepiandrosterone (DHEA) and DHEA sulphate (DHEAS) is poorly understood. It depends in a large part on their transformation into testosterone and estradiol. The capacity of DHEA as a neurosteroid, the recent discovery of putative specific DHEA receptors on endothelial and vascular smooth muscle cells, the steady decrease of DHEA production from the 40s on, together with certain human epidemiologic data as well as various beneficial effects of DHA supplementation in rodents have suggested the possibility that this steroid is involved in cognitive and memory, metabolic and vascular, immune and sexual functions and in their aging. However, epidemiologic studies are conflicting, and no well-designed clinical trials have definitely substantiated the role of DHEA in these functions in humans, or the utility and safety of DHEA supplementation. However, beneficial effects seem plausible in women with several conditions according to the results of double-blind placebo-controlled trials: the dose of 30 to 50 mg seems beneficial to the mood, sense of well being and sexual desire and activity of women with adrenal insufficiency. The only long-term trial of supplementation devoted to women over 60 reported significant increases in bone mineral density and, in the 70-79-year-old subgroup, in sexual desire, arousal, activity and satisfaction. The dose of 200 mg also proved to decrease disease activity in systemic lupus erythematosus. Lastly, high DHEA doses have improved mood in various groups of patients of any age and gender with depressive symptoms. The use of DHEA therapy may also be discussed in women of any age when a trial of androgen supplementation seems justified because of the existence of an inhibited sexual desire or a sexual arousal disorder associated with documented androgen deficiency. The rather weak conversion of DHEA into testosterone protects from the risk of overdosing associated with testosterone preparations. However, it must be realized that DHEA is also converted into estradiol, which may be a risk factor for breast or endometrial cancer in postmenopausal women. Unlike women, no consistent beneficial effect has been found for men in the placebo-controlled trials. The present data do not exclude a role of DHEA in other conditions, but this remains to be properly established. This paper includes practical considerations on dosage to be used, contraindications and follow-up. PMID- 14551725 TI - MRI of cardiac rhabdomyoma in the fetus. AB - Primary cardiac tumors are rarely diagnosed in utero and are usually seen on prenatal echocardiography. Cardiac rhabdomyomata can be associated with tuberous sclerosis. Prenatal MRI can be performed to assess associated malformations. This case report illustrates the ability of fetal MRI to image cardiac rhabdomyata and compares it with prenatal and postnatal echocardiography. PMID- 14551727 TI - Patient radiation exposure in uterine artery embolization of leiomyomata: calculation of organ doses and effective dose. AB - The goal of this study was estimation of patient effective dose from uterine artery embolization of leiomyomata. Parameters and data relevant to patient dose were recorded for 33 consecutive procedures. Using Monte Carlo simulation of radiation transport, organ and effective doses were calculated in detail for a subset of five procedures, to estimate the effective dose for all procedures. Mean dose area product was 59.9, median 23.4, and range 8.8-317.5 Gycm(2). Mean absorbed ovarian dose was calculated as 51 mGy in the five procedures. Using the dose conversion factor estimated from the Monte Carlo simulation for all procedures a mean estimated effective dose of 34 mSv (median 13 mSv, range 5-182 mSv) results, with a tendency to lower values regarding the succession of the procedures. Patients' radiation exposure level is up to twice of that of an abdominal CT examination. Angiographic equipment related dose-reducing features and radiographic technique essentially influence organ doses and effective dose. Consistent application of dose-reducing techniques and awareness of radiation exposure justifies uterine artery embolization as a therapeutic option for the treatment of uterine fibroids. PMID- 14551726 TI - Ultrathin needle (25 G) aspiration lung biopsy: diagnostic accuracy and complication rates. AB - The aim of this study was to evaluate the diagnostic accuracy and complication rate of 25-G fine-needle aspiration biopsy (FNAB) of the lung in patients with suspected malignant focal lesions and abnormal lung function. The 25-G FNAB was performed in 123 patients who underwent prebiopsy CT and pulmonary function tests. Retrospective evaluation included pulmonary function, cytology, size of the lesion, depth of location, presence of emphysema on CT, needle passes, pneumothorax and drainage. The final diagnosis (gold standard) was based on histopathology after surgical resection or follow-up and response to treatment. Sixty-one patients had normal lung function or mild impairment (group 1) and 62 had moderate or severe impairment (group 2). Pneumothorax occurred in 26 of 126 procedures (20.6%) with drainage needed in 11 (8.7%). In group 2 pneumothorax occurred in 19 of 63 procedures (30.15%) with drainage needed in 11 (17.5%). The sensitivity, specificity and diagnostic accuracy of cytology results were 93.6, 100 and 94.4%, respectively. FEV1 ( p=0.014), FEV1/FVC ( p=0.005), FEF25-75 ( p=0.001), DLCO ( p=0.013) and presence of emphysema on CT ( p<0.001) correlated with pneumothorax (Student's t test). The 25-G lung FNAB is accurate and safe in diagnosing malignancy in patients with severe lung functional abnormality. Patients with moderate to severe airway obstruction have a higher prevalence of pneumothorax than patients with mild or no functional impairment. PMID- 14551729 TI - The combined impact of pH and activated carbon on the elemental composition of a liquid conifer embryogenic tissue initiation medium. AB - This study considered the elemental composition of plant tissue culture media in response to pH and two different types of activated C (AC; tissue culture and non acid-washed grades) in liquid media. When tissue culture medium is supplemented with AC the method of AC addition and pH adjustment can greatly impact the final medium pH, in turn, altering mineral availability. Over the pH range of 4-7, Cu and Zn adsorbed (95% and 50%) onto the two physically different ACs to the same extent. As the pH exceeded 5.8, precipitation became pronounced, resulting in 50% reductions in Mn and Fe and smaller reductions in Ca (20%), and P (15%), independent of AC. Non acid-washed AC released significant levels of Mg (65% increase) and Ca (10% increase) at pH 5.8 compared to the no-AC control. No adsorption was indicated for inorganic anions. Low levels for Cu and Zn are a concern when using AC, and low levels of Fe and Mn are a concern when the pH of the medium exceeds 5.8. Due to its impurity content and difficulty associated with its neutralization, non-acid-washed AC may be a poor choice for use in tissue culture medium. PMID- 14551728 TI - Focal nodular hyperplasia in normal and fatty liver: a qualitative and quantitative evaluation with contrast-enhanced ultrasound. AB - The aim of this study was to describe gray-scale appearance of liver parenchyma and focal nodular hyperplasia (FNH) by pulse inversion (PI) ultrasound (US) at baseline and after contrast agent administration in patients with normal and fatty liver. Sixteen consecutive patients (12 women, 4 men) with 29 previously diagnosed FNHs (15 of 29 located in normal liver and 14 of 29 in fatty liver) underwent PI US before and after SH U 508A (Levovist) injection. Signal intensity values were measured within the FNHs and the adjacent liver parenchyma in selected images. Baseline echogenicity of fatty liver was higher (15.19 +/- 2.90 dB +/- SD) than normal liver (10.91 +/- 3.15 dB +/- SD; p<0.001). After Levovist administration, normal livers (7 of 16) showed a statistically significant increase of echogenicity (16.59 +/- 3.81 dB +/- SD; p<0.001) in comparison with fatty livers (9 of 16; 15.75 +/- 3.12 dB +/- SD). The FNHs located in normal liver showed baseline echogenicity higher (12.29 +/- 3.22 dB +/- SD) than that of FNHs arising in fatty liver (7.06 +/- 2.43 dB +/- SD; p<0.001). After Levovist administration, FNHs located in normal liver showed a statistically significant increase of echogenicity (25.30 +/- 4.62 dB +/- SD) in comparison with FNHs located in fatty liver (13.58 +/- 3.54 dB +/- SD; p<0.001); the latter always showed mean values of echogenicity lower than surrounding liver parenchyma. In our series decreased contrast-enhancement pattern of both fatty liver and FNHs located in fatty liver was the most prominent finding when Levovist is administered. Contrast washout was a distinctive feature of FNH arising from the fatty liver. PMID- 14551730 TI - Synthesis of an HIV-1 Tat transduction domain-rotavirus enterotoxin fusion protein in transgenic potato. AB - A DNA fragment encoding a 12-amino acid (aa) HIV-1 Tat transduction peptide fused to a 90-aa murine rotavirus NSP4 enterotoxin protein (Tat-NSP4(90)) was transferred to Solanum tuberosum by Agrobacterium tumefaciens-mediated transformation. The fusion gene was detected in the genomic DNA of transformed plant leaf tissues by PCR DNA amplification. The Tat-NSP4(90 )fusion protein was identified in transformed tuber extracts by immunoblot analysis using anti NSP4(90) and anti-Tat as the primary antibodies. Enzyme-linked immunosorbent assay results showed that the Tat-NSP4(90) fusion protein made up to 0.0015% of the total soluble tuber protein. The synthesis of Tat-NSP4(90) fusion protein in transformed potato tuber tissues demonstrates the feasibility of plant cell delivery of the HIV-1 Tat transduction domain as a carrier for non-specific targeting of fused antigens to the mucosal immune system. PMID- 14551731 TI - Stable transformation of rice (Oryza sativa L.) via microprojectile bombardment of highly regenerative, green tissues derived from mature seed. AB - A highly efficient and reproducible transformation system for rice ( Oryza sativa L. cv. Taipei 309) was developed using microprojectile bombardment of highly regenerative, green tissues. These tissues were induced from mature seeds on NB based medium containing 2,4-dichlorophenoxyacetic acid (2,4-D), 6 benzylaminopurine (BAP) and high concentrations of cupric sulfate under dim light conditions; germinating shoots and roots were completely removed. Highly regenerative, green tissues were proliferated on the same medium and used as transformation targets. From 431 explants bombarded with transgenes [i.e. a hygromycin phosphotransferase ( hpt) gene plus one of a wheat thioredoxin h ( wtrxh), a barley NADP-thioredoxin reductase ( bntr), a maize Mutator transposable element ( mudrB) or beta-glucuronidase ( uidA; gus) gene], 28 independent transgenic events were obtained after an 8- to 12-week selection period, giving a 6.5% transformation frequency. Of the 28 independent events, 17 (61%) were regenerable. Co-transformation of the second introduced transgene was detected in 81% of the transgenic lines tested. Stable integration and expression of the foreign genes in T(0) plants and T(1) progeny were confirmed by DNA hybridization, western blot analyses and germination tests. PMID- 14551732 TI - Production of a fusion protein consisting of the enterotoxigenic Escherichia coli heat-labile toxin B subunit and a tuberculosis antigen in Arabidopsis thaliana. AB - Transgenic plants are potentially safe and inexpensive vehicles to produce and mucosally deliver protective antigens. However, the application of this technology is limited by the poor response of the immune system to non particulate, subunit vaccines. Co-delivery of therapeutic proteins with carrier proteins could increase the effectiveness of the antigen. This paper reports the ability of transgenic Arabidopsis thaliana plants to produce a fusion protein consisting of the B subunit of the Escherichia coli heat-labile enterotoxin and a 6 kDa tuberculosis antigen, the early secretory antigenic target ESAT-6. Both components of the fusion protein were detected using GM1-ganglioside-dependent enzyme-linked immunosorbant assay. This suggested the fusion protein retained both its native antigenicity and the ability to form pentamers. PMID- 14551734 TI - Genetic transformation and regeneration of rubber tree (Hevea brasiliensis Muell. Arg) transgenic plants with a constitutive version of an anti-oxidative stress superoxide dismutase gene. AB - Agrobacterium tumefaciens-mediated genetic transformation and the regeneration of transgenic plants was achieved in Hevea brasiliensis. Immature anther-derived calli were used to develop transgenic plants. These calli were co-cultured with A. tumefaciens harboring a plasmid vector containing the H. brasiliensis superoxide dismutase gene (HbSOD) under the control of the CaMV 35S promoter. The beta-glucuronidase gene (uidA) was used for screening and the neomycin phosphotransferase gene (nptII) was used for selection of the transformed calli. Factors such as co-cultivation time, co-cultivation media and kanamycin concentration were assessed to establish optimal conditions for the selection of transformed callus lines. Transformed calli surviving on medium containing 300 mg l(-1) kanamycin showed a strong GUS-positive reaction. Somatic embryos were then regenerated from these transgenic calli on MS2 medium containing 2.0 mg l(-1) spermine and 0.1 mg l(-1) abscisic acid. Mature embryos were germinated and developed into plantlets on MS4 medium supplemented with 0.2 mg l(-1) gibberellic acid, 0.2 mg l(-1) kinetin (KIN) and 0.1 mg l(-1) indole-3-acetic acid. A transformation frequency of 4% was achieved. The morphology of the transgenic plants was similar to that of untransformed plants. Histochemical GUS assay revealed the expression of the uidA gene in embryos as well as leaves of transgenic plants. The presence of the uidA, nptII and HbSOD genes in the Hevea genome was confirmed by polymerase chain reaction amplification and genomic Southern blot hybridization analyses. PMID- 14551733 TI - Induced androgenesis in tomato (Lycopersicon esculentum Mill.). III. Characterization of the regenerants. AB - We present data on the morphological, cytological, biochemical and genetic characteristics of tomato regenerants obtained through anther culture. As a result of induced androgenesis, more than 6,000 rooted regenerants were developed that differed both from the donor plants and among each other with respect to habitus and leaf, flower and inflorescence morphology. Cytological analysis revealed a great variability in chromosome number in the cells of the regenerated plants. While most of the regenerants were mixoploid, the majority of the cells had a haploid chromosome number. R(1) and R(2) progenies were tested for their resistance to Clavibacter michiganense subsp. michiganense ( Cmm 7). Some of the regenerants were resistant to the pathogen. A biochemical analysis of fruit from R(3) and R(4) plants showed a higher content of dry matter, sugars and vitamin C in the regenerant plants obtained from the hybrids than in those from the cultivars and control plants. The values of the parameters of hybrid regenerants grown in the greenhouse were about 1.5-fold higher than those of the hybrid regenerants grown in the field, and this trend is clearly expressed in all of the hybrid regenerants. The results obtained suggest that induced androgenesis and gametoclonal variation may be used as an additional tool to create a large range of new forms. The application of the latter in breeding programs would accelerate the development of tomato lines and varieties that would be more productive, disease-resistant, highly nutritive and flavour-acceptable. PMID- 14551735 TI - Plasma and cerebrospinal fluid pharmacokinetics of SU5416 after intravenous administration in nonhuman primates. AB - PURPOSE: SU5416 is a small, lipophilic synthetic molecule that selectively inhibits the tyrosine kinase activity of the VEGF receptor Flk-1/KDR. The role of this agent in brain tumors is currently being investigated. Pharmacokinetic studies of SU5416 have been performed in humans; however, there have been no studies of its penetration in the cerebrospinal fluid (CSF). We studied the pharmacokinetics of SU5416 in plasma and CSF after intravenous (i.v.) administration using a nonhuman primate model that is highly predictive of the CSF penetration in humans. EXPERIMENTAL DESIGN: SU5416 (85 mg/m(2), about 3.8 mg/kg) was administered i.v. over 20 min to four nonhuman primates. Serial plasma and CSF samples were obtained prior to, during, and after completion of the infusion for determination of SU5416 concentrations. SU5416 was measured in plasma and CSF using high-performance liquid chromatography (HPLC). Concentration versus-time data were modeled using model-independent and model-dependent methods. RESULTS: Peak plasma concentrations ranged from 6.3 to 14.5 microM and the mean plasma AUC was 620+/-180 microM.min. Disappearance of SU5416 from the plasma was best described by a one-compartment model with a half-life of 39+/-2.9 min. The volume of distribution was 36+/-11 l/m(2) and the clearance was 0.62+/ 0.2 l/min per m(2). SU4516 was not quantifiable in the CSF. CONCLUSIONS: There is minimal penetration of SU5416 into the CSF after i.v. administration. The very low CNS exposure to SU5416 after i.v. dosing suggests that this agent is not optimal for the treatment of leptomeningeal tumors. PMID- 14551736 TI - Antitumoral properties of two new vanadyl(IV) complexes in osteoblasts in culture: role of apoptosis and oxidative stress. AB - BACKGROUND: Vanadium derivatives have been reported to display different biological effects, and in particular antineoplastic activity has been demonstrated in both in vivo and in vitro studies. PURPOSE. To study the effect of two new organic vanadyl(IV) complexes (one with glucose, GluVO, and the other with naproxen, NapVO) in osteosarcoma cells. METHODS: UMR106 osteosarcoma cells and, for comparison, nontransformed MC3T3E1 osteoblasts were used. Proliferation and differentiation were assessed using the crystal violet assay and ALP specific activity, respectively. Morphological alterations were assessed by light microscopy. Lipid peroxidation was evaluated in terms of production of thiobarbituric acid-reactive substances (TBARS) and apoptosis was measured using annexin V. Extracellular regulated kinase (Erk) activation was investigated by Western blotting. RESULTS: Vanadium complexes caused morphological alterations and they strongly inhibited UMR106 cell proliferation and differentiation. In contrast, in MC3T3E1 cells, these vanadium derivatives had a relatively weak action. In UMR106 tumoral cells there was a significant increase in TBARS production. Both vanadium complexes induced apoptosis and activation of Erk. PD98059, an inhibitor of Erk phosphorylation, did not block the vanadium-induced antitumoral action. However, the antioxidants vitamins C and E abrogated the apoptosis and TBARS production induced by the vanadium complexes. CONCLUSIONS: GluVO and NapVO exerted an antitumoral effect in UM106 osteosarcoma cells. They inhibited cell proliferation and differentiation. While the Erk cascade seems not to be directly related to the bioactivity of these vanadium derivatives, the action of both vanadium complexes with organic ligands may be mediated by apoptosis and oxidative stress. PMID- 14551737 TI - High dose methylprednisolone can induce remissions in CLL patients with p53 abnormalities. AB - Abnormalities of the p53 gene are known to confer detrimental effects in chronic lymphocytic leukaemia (CLL) and are associated with short survival. We have used high dose methylprednisolone (HDMP) to treat 25 patients with advanced refractory CLL of whom 45% had p53 abnormalities shown by one or more methods: flow cytometry, fluorescent in situ hybridisation and direct DNA sequencing. Fifteen were resistant to fludarabine and 16 were non-responders to their most recent therapy. Methylprednisolone had a cytotoxic effect on lymphocytes from 95% of cases assessed by an ex vivo apoptotic drug sensitivity index (DSI). HDMP was given alone or in combination with other drugs: vincristine, CCNU, Ara-C, doxorubicin, mitoxantrone and chlorambucil, according to the results of DSI. Three patients were treated twice and each treatment was analysed separately. The overall response rate was 77% with a median duration of 12 months (range 7 -23+). Responders included 5/10 with abnormal p53, of which two achieved nodular PR. Patients with p53 abnormalities fared worse than those with normal p53. There were no differences in response according to whether HDMP was used alone or in combination. Nine of the 22 evaluable patients (3 NR and 6 PR) have died from progressive disease or transformation. Main toxicity was infection in 7/25 patients. Event free and overall survival were significantly better in responders vs non-responders ( P>0.0001 and P=0.04 respectively). Patients with a DSI of 100% to steroids had a better overall and event free survival, but this was not statistically significant. This study demonstrates that HDMP alone or in combination with other agents is a useful treatment strategy in refractory CLL including patients with p53 abnormalities. PMID- 14551738 TI - A novel chromosomal translocation t(3;7)(q26;q21) in myeloid leukemia resulting in overexpression of EVI1. AB - The EVI1 proto-oncogene encodes a nuclear zinc finger protein that acts as a transcription repressor factor. In myeloid leukemia it is often activated by chromosomal rearrangements involving band 3q26, where the gene has been mapped. Here we report two leukemia cases [a chronic myeloid leukemia blast crisis (CML BC) and an acute myeloid leukemia (AML) M4] showing a t(3;7)(q26;q21) translocation in a balanced and unbalanced form, respectively. Fluorescent in situ hybridization (FISH) analysis revealed that both patients showed a breakpoint on chromosome 3 inside the clone RP11-33A1 containing the EVI1 oncogene and, on chromosome 7, inside the clone RP11-322M5, partially containing the CDK6 oncogene which is a D cyclin-dependent kinase gene, observed to be overexpressed and disrupted in many hematological malignancies. Reverse transcriptase polymerase chain reaction (RT-PCR) analysis showed overexpression of EVI1 in both cases, but excluded the presence of any CDK6/ EVI1 fusion transcript. CDK6 expression was also detected. Together, these data indicate that EVI1 activation is likely due not to the generation of a novel fusion gene with CDK6 but to a position effect dysregulating its transcriptional pattern. PMID- 14551739 TI - Reduced incidence of acute graft versus host disease (GVHD) of the gut in Chinese carriers of Helicobacter pylori during allogeneic bone marrow transplantation. AB - Helicobacter pylori ( H. Pylori) infection is associated with gastritis and peptic ulcer, but its relationship with gut graft versus host disease (GVHD) is unknown. We investigated the association between H. Pylori carriage and incidence and severity of mucosal toxicity and GVHD in 128 consecutive matched sibling stem cell transplantation (SCT) recipients. Using a verified enzyme linked immunosorbant assay (ELISA), 43.5% of patients had H. Pylori exposure before SCT. There was absolute concordance between serological and breath test data in 40 prospective cases. There was no increased risk in WHO grade 3 or 4 mucositis in H. Pylori carriers. Significant (grade II or above) overall GVHD was only predicted by preceding mucositis (p<0.001), while gut GVHD was associated with increased age (p=0.001) and mucositis (p=0.022). Despite increased incidence with age, H. Pylori carriage was associated with significantly reduced risk of gut GVHD (p=0.04) but not overall GVHD. The reduced risk of immune-mediated gut inflammation in H. Pylori carriers after SCT may be related to the known reduced incidence of inflammatory bowel disease in chronic H. Pylori carriers. PMID- 14551741 TI - Gamma-irradiation of blood products following autologous stem cell transplantation: surveillance of the policy of 35 centers. AB - Blood products should be irradiated during allogeneic stem cell transplantation and before performing autologous stem cell harvest for prevention of acute transfusion-associated graft-versus-host disease (TA-GVHD). Usually, irradiation of all blood products is continued lifelong in the allogeneic setting. Up to now, no broadly accepted rules exist concerning autologous stem cell transplantation. We present here the results of an inquiry sent to 47 German transplantation centers regarding the transfusion policy following autologous stem cell transplantation. The results of 35 answering centers are included. Ten out of 35 centers offer irradiated blood products lifelong to their patients, mainly for the prevention of mistransfusion of non-irradiated blood components to allogeneic recipients. Twenty-two out of 35 centers administer irradiated blood products for a special time span after autologous stem cell transplantation. In most centers, this time span is from 3 to 6 months. Only few centers (4/35) expand this time span to 1-2 years after transplantation. A minority of centers (3/35) gave non irradiated blood products to all of their patients or to patients not suffering from acute leukemia or after total body irradiation (TBI) containing preparative regimens. Most centers (19/35) deliver irradiated blood products irrespective of the conditioning regimen. Fifty-three percent of the centers decide to donate irradiated blood products not depending on immunological reconstitution. But in most centers some kind of hematological reconstitution is a major criterion for termination of irradiated blood products. Sixty-four percent of the centers made no difference in transfusion policy in regard to the underlying disease. No center experienced cases of proofed TA-GVHD. Guidelines should be worked out concerning transfusion policy after autologous stem cell transplantation. PMID- 14551742 TI - Evaluation of serum ferritin in screening for iron deficiency in tuberculosis. AB - Serum ferritin (SF) values 0.05). Calcium overload occurred prior to an abrupt reduction in ADC in the peripheral ischaemic area. The relative ADC (rADC) and FI (rFI) were negatively correlated in the frontoparietal cortex ( r=-0.9, P <0.001), but not in the caudoputamen ( r=-0.21, P >0.05). Our findings suggest that ADC of the perifocal ischaemic area might reflect the changes in intracellular calcium which occur in early ischaemia. They may also suggest that, once the calcium level is high enough and infarction ensues, changes in ADC may not induce a further rise in calcium concentration. PMID- 14551760 TI - Normalized regional brain atrophy measurements in multiple sclerosis. AB - There is still a controversy regarding the best regional brain atrophy measurements in multiple sclerosis (MS) studies. The aim of this study was to establish whether, in a cross-sectional study, the normalized measurements of regional brain atrophy correlate better with the MRI-defined regional brain lesions than the absolute measurements of regional brain atrophy. We assessed 45 patients with clinically definite relapsing-remitting (RR) MS (median disease duration 12 years), and measured T1-lesion load (LL) and T2-LL of frontal lobes and pons, using a reproducible semi-automated technique. The regional brain parenchymal volume (RBPV) of frontal lobes and pons was obtained by use of a computerized interactive program, which incorporates semi-automated and automated segmentation processes. A normalized measurement, the regional brain parenchymal fraction (RBPF), was calculated as the ratio of RBPV to the total volume of the parenchyma and the cerebrospinal fluid (CSF) in the frontal lobes and in the region of the pons. The total regional brain volume fraction (TRBVF) was obtained after we had corrected for the total volume of the parenchyma and the CSF in the frontal lobes and in the region of the pons for the total intracranial volume. The mean coefficient of variation (CV) for RBPF of the pons was 1% for intra observer reproducibility and 1.4% for inter-observer reproducibility. Generally, the normalized measurements of regional brain atrophy correlated with regional brain volumes and disability better than did the absolute measurements. RBPF and TRBVF correlated with T2-LL of the pons (r=-0.37, P=0.011, and r= -0.40, P=0.0005 respectively) and with T1-LL of the pons (r=-0.27, P=0.046, and r=-0.31, P=0.04, respectively), whereas RBPV did not (r=-0.18, P = NS). T1-LL of the frontal lobes was related to RBPF (r=-0.32, P=0.033) and TRBVF (r=-0.29, P=0.05), but not to RBPV (R=-0.27, P= NS). There was only a trend of correlation between T2-LL of the frontal lobes and RBPF (r=-0.27, P=0.06) and TRBVF (r=-0.28, P=0.057), and no correlation with RBPV (r=-0.23, P= NS). The magnitude of correlation between the expanded disability status scale (EDSS) and pontine and frontal lobe RBPF and TRBVF was more than twice as high as the correlation between EDSS and RBPV of the same regions. These data suggest that normalized regional brain atrophy measurements are preferable to absolute regional measurements in cross-sectional studies. PMID- 14551761 TI - Venous variations in the region of the third ventricle: the role of MR venography. AB - In this study, we examined the anatomical variations of the subependymal veins in the region of the foramen of Monro and the third ventricle by MR time of flight (TOF) venography. Fifty healthy subjects, ten patients with third-ventricle tumors, and four patients with lateral-ventricle tumors were included in the study. The courses of the anterior septal vein (ASV), thalamostriate vein (TSV), and internal cerebral vein (ICV) were studied. The proximity of the venous angle, the false venous angle, and the ASV-ICV junction to the posterior margin of the foramen of Monro was measured. In 69 (53.9%) sides, the ASV-ICV junction was located at the venous angle and at the posterior margin of the foramen of Monro. In 59 (46.1%) sides, the ASV-ICV junction was located beyond the foramen of Monro. Our study shows the high incidence of posteriorly located ASV-ICV junctions, which can be crucial in the planning of a better surgical approach. We strongly recommend that MR venography, which is a short radiological examination, be used before one operates on third-ventricle and lateral-ventricle tumors. PMID- 14551762 TI - Cloning and characterization of leaf cDNAs that are differentially expressed between wheat hybrids and their parents. AB - Previous studies have shown that heterosis is associated with differential gene expression between hybrids and their parents. In this study, we performed a screen for genes that are differentially expressed between wheat hybrids and their parents in jointing-stage leaves and flag leaves using the differential display technique. Twenty-four differentially expressed cDNA were cloned and sequenced, and their expression patterns were confirmed by reverse-Northern blotting. Sequence analysis and database searches revealed that among the genes that showed differential expression between hybrid and parents were transcription factor genes and genes involved in metabolism, signal transduction, disease resistance, and retrotransposons. These results indicate that hybridization between two parental lines can cause changes in the expression of a variety of genes, and it is concluded that the altered pattern of gene expression in the hybrid may be responsible for the observed heterosis. PMID- 14551763 TI - Transplantation of in vitro-generated Schistosoma mansoni mother sporocysts into Biomphalaria glabrata. AB - Specific studies on schistosome gene functions require both access to the parasite stages, preferably the larvae, and to complete the life cycle. In the present study, we investigated whether short-term in vitro cultivation of sporocysts and surgical transplantation into snails could be combined to produce cercariae. Miracidia were maintained in vitro in the presence of Biomphalaria glabrata embryonic (Bge) cells or, alternatively, in Bge-cell-conditioned medium. The transformation of miracidia to mother sporocysts was observed in both cases. Two day-old sporocysts were transplanted into the cephalopedal sinus of recipient snails. Transplantation efficiencies varied between 16% and 43%, depending on the culture of the sporocysts in terms of the number of cercariae producing snails. Cercariae recovered from these snails were used to successfully infect hamsters, demonstrating that short term in vitro-generated sporocysts undergo normal cercariogenesis following transplantation. This combination of in vitro cultivation and transplantation may be useful for novel experimental approaches to investigate the genes involved in larval development or host-parasite molecular interactions. PMID- 14551764 TI - How accurately can QT interval be measured in newborn infants? AB - We determined inter- and intra-observer variability of measurements of QT and heart rate corrected QT (QTc) interval in newborn infants. In 55 newborns, a standard 12-lead electrocardiogram (ECG) with rhythm strip was analysed independently by three observers. QT and the preceding RR interval were measured in three different beats. The QTc interval was calculated using the Bazett formula (QTc = QT/ radical RR). Observers repeated their measurements after the recruitment period blinded for their first reading. Inter- and intra-observer variability were determined using the Bland and Altman method. In total, 59 ECGs were recorded. Due to artifacts, six had to be excluded. A marked inter-observer variability was observed in the measurement of the QT and QTc interval. Important intra-observer variability was denoted, with differences up to 80 ms for QTc. CONCLUSION: mainly due to high heart rates, important inter- and intra-observer variability hampers a correct interpretation of the QT interval in newborn infants. We suggest not to rely on one single measurement and to apply different methods of measurement and a margin of safety when interpreting QT and heart rate corrected QT intervals in these age groups for making therapeutic decisions. PMID- 14551765 TI - Blink-related sensorimotor anatomy in the rat. AB - Protection of the eye and maintenance of the precorneal tear film depend on sensory innervation of the cornea and eyelids and motor innervation of muscles involved in closing and opening the eyes. Using a variety of fluorescent and transganglionic tracers, the sensorimotor innervation of blink-related orbital and periorbital structures was studied in Sprague-Dawley rats. The orbicularis oculi muscle surrounded the entire palpebral fissure and was innervated by motoneurons located along the dorsal cap of the ipsilateral facial motor nucleus. Upper and lower eyelid orbicularis oculi motoneurons were strictly ipsilateral and co-extensive, but upper eyelid orbicularis oculi motoneurons were, on average, slightly rostral and lateral to lower eyelid orbicularis oculi motoneurons. Facial motoneurons supplying the frontoscutularis, a muscle that helps to elevate the upper eyelid, were located in the medial division of the ipsilateral facial motor nucleus. Presumptive type Abeta afferents from the cornea terminated most prominently at the junction of the first cervical segment and the spinal trigeminal nucleus, pars caudalis. There was a second concentration of corneal terminations at the junction of pars caudalis and pars interpolaris of the spinal trigeminal nucleus. Sparse projections to the spinal trigeminal nucleus, pars oralis and the principal trigeminal nucleus were also detected. Presumptive type Abeta afferents from the eyelids terminated throughout the rostrocaudal extent of the spinal trigeminal nucleus with a heavy concentration within laminae III and IV of the first cervical segment. Presumptive types Adelta and C terminals from the eyelids were virtually limited to laminae I and II of the first cervical segment. Central terminations from the frontal nerve were present in the principal trigeminal nucleus and throughout the spinal trigeminal nucleus, but were most prominent within the dorsal horn of the first cervical segment. Our comprehensive description of blink-related sensorimotor anatomy in rats will provide a foundation for future physiological studies of blinking. PMID- 14551768 TI - Barrett's oesophagus: time for consensus. PMID- 14551766 TI - The role of M cells of human nasopharyngeal lymphoid tissue in influenza virus sampling. AB - Little is known about the role of the M cells of human nasopharyngeal lymphoid tissue in the sampling of viruses that cause respiratory infections. To clarify whether M cells could function as a gateway for influenza virus into human nasopharyngeal lymphoid tissue, excised adenoid tissue was incubated in media containing influenza A virus for 30, 60, and 90 min, respectively. Transmission electron microscopic observation revealed that many influenza viruses adhered to M cell surfaces and were taken up into the cytoplasmic vesicles of M cells after 30 min incubation; the viruses had been transported into enfolded lymphoid cells after 60 min incubation. By staining M cells with Sambucus nigra lectin, which specifically recognizes the NeuAcalpha2,6 Gal linkage of sialoprotein, it was also found that abundant receptors for the human influenza virus are present on the M cell surface. Our findings indicated that M cells of human nasopharyngeal tonsils function as a major port for influenza A virus entry and that the virus could be efficiently transferred to enfolded macrophages and lymphoid cells by M cells. The transport of influenza viruses to lymphoid cells by M cells may promote antigen delivery to the immune system, and these findings may be important for systemic delivery of those influenza viruses that have the capacity to productively infect cells outside of the respiratory tract. PMID- 14551769 TI - Hematopoietic organs of Manduca sexta and hemocyte lineages. AB - Cells of the moth immune system are derived from organs that loosely envelop the four wing imaginal discs. The immune response in these insects is believed to depend on the activities of two main classes of hemocytes: plasmatocytes and granular cells. The fates of cells that arise from these hematopoietic organs have been followed by immunolabeling with plasmatocyte-specific and granular-cell specific antibodies. Cells within each hematopoietic organ differ in their coherence and in their expression of two plasmatocyte-specific surface proteins, integrin and neuroglian. Within an organ there is no overlap in the expression of these two surface proteins; neuroglian is found on the surfaces of the coherent cells while integrin is expressed on cells that are losing coherence, rounding up, and dispersing. A granular-cell-specific marker for the protein lacunin labels the basal lamina that delimits each organ but only a small number of granular cells that lie on or near the periphery of the hematopoietic organ. When organs are cultured in the absence of hemolymph, all cells derived from hematopoietic organs turn out to immunolabel with the plasmatocyte-specific antibody MS13. The circulating plasmatocytes derived from hematopoietic organs have higher ploidy levels than the granular cells and represent a separate lineage of hemocytes. PMID- 14551770 TI - Expression profiling of homeobox genes in silk gland development in the mulberry silkworm Bombyx mori. AB - The homeobox (Hox) genes constitute an evolutionarily conserved family encoding transcription factors which play major roles in segmental identity and organ specification, across species. The expression patterns of three Hox genes, Antennapedia (Antp), Ultrabithorax (Ubx) and even-skipped (eve) were analyzed during silk gland development in Bombyx mori. Antp followed a middle silk gland (MSG) restricted pattern of expression, whereas Ubx was specifically expressed in the posterior silk gland (PSG) during embryonic and larval developmental stages. Eve protein, on the other hand, was expressed in both MSG and PSG. We have also identified and characterized a novel Pax-like mRNA that is expressed exclusively in the PSGs during embryonic and larval development. The expression of Antp, Ubx and Pax-like reached maximum levels in the fifth larval intermoult and no expression was detected during the intervening moults. The region-specific expression of certain Hox genes appears to be responsible for the specification of silk gland compartments, whereas other Hox genes may play a global role in controlling the expression of genes encoding silk proteins. PMID- 14551771 TI - Distinct MADS-box gene expression patterns in the reproductive cones of the gymnosperm Gnetum gnemon. AB - Expression patterns from in situ hybridization of four MADS-box genes (GGM7, GGM9, GGM11, and GGM15) from the gymnosperm species Gnetum gnemon are presented. Together with previously published data about putative orthologs of floral homeotic genes from G. gnemon (GGM2, GGM3, GGM13), we describe seven temporally and spatially distinct expression patterns in male, female or both types of reproductive units which very likely reflect the diversity of MADS-box gene function in gymnosperm cones. There is evidence that some aspects of the observed differential expression have been conserved since the last common ancestor of extant angiosperms and gymnosperms about 300 million years ago. PMID- 14551772 TI - Morphological development of anthers induced by the dimorphic smut fungus Microbotryum violaceum in female flowers of the dioecious plant Silene latifolia. AB - When inoculated with the dimorphic smut fungus Microbotryum violaceum (Pers.) G. Deml and Oberwinkler, the female flower of the dioecious plant Silene latifolia (Miller) E.H.L. Krause develops anther-like structures filled with spores instead of pollen grains. Using natural scanning electron microscopy, Nomarski interference microscopy, and fluorescence microscopy, we investigated the morphological modifications of the host plant resulting from this parasitism and the localization of smut hyphae in the flower bud. Flowers of infected plants lasted significantly longer than those of healthy plants, probably because the infection strengthened floral organs, such as the flower base and the anther filaments. Smut hyphae were observed throughout all organs of the young flower buds of infected plants, including sepals, petals, stamens, and pistil primordia. In healthy female flowers, anthers initiated sporogenous cell formation, but lacked parietal cell layers. By contrast, the parietal cell layers of infected female flowers differentiated into tapetal tissue, middle cell layers, and endothecial layers, as in the anthers of healthy male flowers. Smut spore formation in the infected anther was initiated in intercellular regions between the sporogenous cells, resulting in degeneration of premature sporogenous cells, tapetal tissue, and middle cell layers. The development of the endothecial layers and epidermis in the infected anther were morphologically normal. PMID- 14551774 TI - Pflugers Archiv and the advent of modern electrophysiology. From the first action potential to patch clamp. AB - This short review recollects the many essential milestones in electrophysiology that were published in Pflugers Archiv. These involve the first measurement of an action potential by J. Bernstein, the requirement of Na+ for the generation of excitation, the prediction of a lipoid membrane surrounding cells by E. Overton, the physical explanation of the resting membrane potential by J. Bernstein, the first detailed description of the conductance properties of excitable tissues by L. Hermann, and more recently the publication of the patch-clamp method by E. Neher and B. Sakmann. PMID- 14551773 TI - Faster oxygen uptake kinetics at the onset of submaximal cycling exercise following 4 weeks recombinant human erythropoietin (r-HuEPO) treatment. AB - We tested the hypothesis that prolonged administration of moderate doses of recombinant human erythropoietin (r-HuEPO) accelerates the initial rate of rise in pulmonary O2 uptake (VO2) in response to submaximal exercise and increases the maximal rate of O2 uptake (VO(2,max)). Sixteen endurance-trained athletes were divided into two groups: r-HuEPO- (n=9) or placebo-treated (n=7). r-HuEPO or placebo (saline) injections were given s.c. 3 times a week for 4 weeks. Exercise testing, before and after the 4 weeks, comprised incremental maximal tests and several transitions from rest to 10-min cycling exercise at 65% VO(2,max). VO2 was measured breath-by-breath during all tests. In the r-HuEPO group, resting haemoglobin concentration (+9.6%) and haematocrit (+8.3%), as well as VO(2,max) (+7.0%) and power output (+7.2%) increased significantly (P<0.05) after the 4 weeks, whereas no change was observed in the control group. The time constant of the primary VO2 response was significantly faster (+18%) after the 4 weeks r HuEPO treatment than before (mean+/-SD; 29.3+/-4.5 vs. 35.7+/-7.4 s, respectively, P<0.05) but was unaffected in the placebo group (34.5+/-7.3 vs. 33.4+/-7.9 s). Collectively, our findings suggest that r-HuEPO contributes both to an acceleration of the dynamic response of VO2 to submaximal exercise and to an increase in maximal exercise capacity. PMID- 14551775 TI - Dual effect of temperature on the human epithelial Na+ channel. AB - The amiloride-sensitive epithelial sodium channel (ENaC) is the rate-limiting step for sodium reabsorption in the distal segments of the nephron, in the colon and in the airways. Its activity is regulated by intracellular and extracellular factors but the mechanisms of this regulation are not yet completely understood. Recently, we have shown that the fast regulation of ENaC by the extracellular [Na+], a phenomenon termed self-inhibition, is temperature dependent. In the present study we examined the effects of temperature on the single-channel properties of ENaC. Single-channel recordings from excised patches showed that the channel open probability (Po, estimated from the number of open channels N.Po, where N is the total number of channels) increased on average two- to threefold while the single-channel conductance decreased by about half when the temperature of the perfusion solution was lowered from approximately 30 to approximately 15 degrees C. The effects of temperature on the single-channel conductance and Po explain the changes of the macroscopic current that can be observed upon temperature changes and, in particular, the paradoxical effect of temperature on the current carried by ENaC. PMID- 14551776 TI - Coactivation pattern in human quadriceps during isokinetic knee-extension by muscle functional MRI. AB - We tested activation patterns of individual muscles and neuromuscular compartments (NMCs) of the quadriceps femoris (QF) during knee-extension exercises (KEE). Eight healthy men performed repetitive isokinetic KEE at 120 degrees s(-1). Before and after KEE, muscle functional magnetic resonance images of the thigh were taken to calculate transverse relaxation times (T2) for the individual muscles of the QF. In terms of exercised T2, a moderate correlation was observed between the rectus femoris (RF) and vastus medialis (VM) ( r=0.493, P<0.001), and between the vastus lateralis (VL) and VM ( r=0.347, P<0.01). Furthermore, a strong correlation was also observed between the vastus intermedius (VI) and VM ( r=0.719, P<0.0001), and between the VL and VI ( r=0.783, P<0.0001). There was a significant difference in T2 between the proximal and distal NMCs of the RF. We conclude that at least two strategies are applied for force production in the human QF: (1) coactivation of specific pairs of individual muscles and (2) regional-specific activation in NMCs in the RF. PMID- 14551778 TI - Sweat lactate response between males with high and low aerobic fitness. AB - Sweat lactate indirectly reflects eccrine gland metabolism. However the potential influence of aerobic fitness on sweat lactate is not well-understood. Six males with high aerobic fitness [peak oxygen consumption ( VO(2)peak): 61.6 (2.5) ml.kg(-1).min(-1)] and seven males with low aerobic fitness [ VO(2)peak: 41.8 (6.4) ml.kg(-1).min(-1)] completed a maximal exertion cycling trial followed on a different day by 60 min of cycling (60 rev.min(-1)) in a 30 degrees C wet bulb globe temperature environment. Intensity was individualized at 90% of the ventilatory threshold ( V(E)/ VO(2) increase with no concurrent V(E)/ VCO(2) increase). Sweat samples were collected from the lumbar region every 10 min and analyzed for lactate concentration. Sweat rate (SR) was significantly greater ( p<0.05) for subjects with a high [1445 (254) ml.h(-1)] versus a low [1056 (261) ml.h(-1)] fitness level. Also, estimated total lactate excretion (SRxmean sweat lactate concentration) was marginally greater ( p=0.2) in highly fit males. However, repeated measures ANOVA showed no significant differences ( p>0.05) between groups for sweat lactate concentration at any time point. Current results show highly fit (vs. low fitness level) males have a greater sweat rate which is consistent with previous literature. However aerobic fitness and subsequent variations in SR do not appear to influence sweat lactate concentrations in males. PMID- 14551777 TI - Plasma glucose, insulin and catecholamine responses to a Wingate test in physically active women and men. AB - The influence of gender on the glucose response to exercise remains contradictory. Moreover, to our knowledge, the glucoregulatory responses to anaerobic sprint exercise have only been studied in male subjects. Hence, the aim of the present study was to compare glucoregulatory metabolic (glucose and lactate) and hormonal (insulin, catecholamines and estradiol only in women) responses to a 30-s Wingate test, in physically active students. Eight women [19.8 (0.7) years] and eight men [22.0 (0.6) years] participated in a 30-s Wingate test on a bicycle ergometer. Plasma glucose, insulin, and catecholamine concentrations were determined at rest, at the end of both the warm-up and the exercise period and during the recovery (5, 10, 20, and 30 min). Results showed that the plasma glucose increase in response to a 30-s Wingate test was significantly higher in women than in men [0.99 (0.15) versus 0.33 (0.20) mmol l( 1) respectively, P<0.05]. Plasma insulin concentrations peaked at 10 min post exercise and the increase between this time of recovery and the end of the warm up was also significantly higher in women than in men [14.7 (2.9) versus 2.3 (1.9) pmol l(-1) respectively, P<0.05]. However, there was no gender difference concerning the catecholamine response. The study indicates a gender-related difference in post-exercise plasma glucose and insulin responses after a supramaximal exercise. PMID- 14551779 TI - The effect of an active warm-up on surface EMG and muscle performance in healthy humans. AB - The effect of an active warm-up on maximal voluntary contraction (MVC), maximal instantaneous power output and surface EMG (sEMG) parameters was studied. Eight volunteers [mean (SD) 22 (4) years] completed two trials on the same day, one control (CO), and the other preceded by an active cycling warm-up (WU) at 70% ventilatory threshold determined by the ventilatory equivalent method. Quadriceps muscle temperature, measured from vastus lateralis with a flexible thermistor, was 33.8 (0.4) degrees C in CO compared to 36.8 (0.5) degrees C in WU ( P<0.05). Aural temperature, measured by an infrared tympanic thermistor, was not different between conditions. Experimental trials consisted of three knee-extension maximum voluntary contractions at a 90 degree angle with simultaneous recording of sEMG from the vastus lateralis, followed by three squat jumps performed on a force platform. SEMGs were analysed in the frequency domain as median frequency (MDF) and in the time domain as root mean square (RMS). MDF was 59.2 (14.1) Hz in CO compared to 67.2 (11.8) Hz in WU ( P<0.05), while RMS was higher in CO compared to WU [0.65 (0.28) mV vs. 0.56 (0.19) mV; P<0.05]. MVC was not different [465.7 (107.6) N vs. 490.1 (117.2) N], whilst instantaneous power output during the squat jump was significantly higher in the WU trial [3324 (866) W vs. 3569 (919) W; P<0.05]. These data show MDF to be altered with an active warm-up, which would relate to a greater conduction velocity. This may translate into faster activation of the muscle fibres, thus partly explaining the increase in power output. PMID- 14551780 TI - Physical activity level measured by doubly labeled water and accelerometry in children. AB - The present study investigated the use of a tri-axial accelerometer, Tracmor2, for the measurement of physical activity in children. Eleven children [age 6.9 (2.2) years, body mass 19.5 (5.3) kg and height 112.3 (14.4) cm] were studied. Total daily energy expenditure (TDEE) was measured using the doubly labeled water method over a 2-week period. In addition, basal metabolic rate (BMR) was determined by the ventilated hood system. Physical activity level (PAL) was defined as TDEE/BMR. Tracmor2 was worn during the same 2-week period throughout waking hours after which average counts per day were calculated. The average counts per day were shown to be highly correlated to PAL values measured by doubly labeled water: PAL = 1.156 x 10(-5) x Tracmor2 average counts day(-1) + 0.978 (r=0.79, P<0.01). In conclusion, Tracmor2 is a valid instrument to measure physical activity in children under free-living conditions. PMID- 14551781 TI - Current internal exposure to pesticides in children and adolescents in Germany: urinary levels of metabolites of pyrethroid and organophosphorus insecticides. AB - AIM: Pesticides are widely used throughout the world, in agriculture to protect crops and in public health to control diseases transmitted by vectors or intermediate hosts. After the prohibition of organochlorines such as DDT, today, mainly pyrethroids and organophosphorus insecticides are used. Whereas many studies have been published on background exposure of the population to organochlorines, data on internal exposure of the population to pyrethroids and organophosphorus insecticides are scarce. Here, we report on internal exposure of children and young people, in an urban area in Germany, to pyrethroids and organophosphorus acids, assessed by the analysis of urinary levels of their corresponding specific metabolites. METHODS: Approximately 673 children and adolescents took part in this voluntary investigation, including 331 children <6 years of age. Their parents stated that they and their children had never used pyrethroids or organophosphorus acids in their homes or for medical reasons. We analysed their spot urine samples for six metabolites of organophosphorus insecticides [dimethyl-phosphate (DMP), diethyl-phosphate (DEP), dimethyl thiophosphate (DMTP), diethyl-thiophosphate (DETP), dimethyl-dithiophosphate (DMDTP) and diethyl-dithiophosphate (DEDTP)] and for four metabolites of pyrethroids [cis-3-(2,2-dibromo-vinyl)-2,2-dimethyl-cyclopropane carboxylic acid (Br2CA), cis-3-(2,2-dichloro-vinyl)-2,2-dimethyl-cyclopropane carboxylic acid (cis-Cl2-CA ), trans-3-(2,2-dichloro-vinyl)-2,2-dimethyl-cyclopropane carboxylic acid (trans-Cl2-CA) and 4-fluoro-3-phenoxy-benzoic acid (F-PBA)] using gas chromatographic methods with mass-selective detection. The limit of detection was 0.1-0.2 microg/l for pyrethroid metabolites and 1 microg/l for metabolites of organophosphorus acids; in DMP it was 5 microg/l. RESULTS: The 95th percentiles of the urinary metabolite concentrations were, in microgrammes per litre, DMP 158, DMTP 180, DMDTP 12, DEP 17, DETP 8, DEDTP <1; Br2CA 0.30, cis-Cl2-CA 0.44, trans-Cl2-CA 1.22, F-PBA 0.30. There were no correlations between urinary metabolite levels and the age of the children. CONCLUSION: Current background levels of internal exposure to pyrethroids and organophosphorus insecticides in children and adolescents in Germany are shown. Exposure to these substances in the general population is thought to occur mainly via residues in the diet. The level of background internal pyrethroid exposure in the children is orders of magnitude lower than the corresponding acceptable daily intake (ADI) values published, but the level of internal organophosphate exposure may reach and even exceed ADI values. This observation demands further investigation. PMID- 14551782 TI - Validation of the multiplex kit genRESMPX-2 for forensic casework analysis. AB - Validation studies were carried out using the commercially available PCR multiplex system genRESMPX-2. In addition to amelogenin, this system comprises the complete set of eight STR systems which are components of the German DNA database established in 1998 by the Federal Criminal Office of Germany (BKA). The minimum amount of template DNA which gave a complete DNA pattern ranged between 100 pg and 200 pg. Mixed samples could clearly be assigned from ratios between 1:5 (ACTBP2) and 1:20 (VWA, FGA). Experimental investigations with different forensic materials, environmental studies, reproducibility and precision data as well as practical casework analysis revealed that the genRESMPX-2 kit can be regarded as a sensitive, reliable and robust multiplex system even in the case of samples containing limited amounts or degraded DNA. PMID- 14551783 TI - Re-evaluation of the RBE of 29 kV x-rays (mammography x-rays) relative to 220 kV x-rays using neoplastic transformation of human CGL1-hybrid cells. AB - Neoplastic transformation of human CGL1-hybrid cells was examined after exposure to 29 kV x-rays (mammography x-rays) and conventional 220 kV x-rays. The study was designed to repeat, under well-defined irradiation and culture conditions, an earlier investigation by Frankenberg et al. (Radiat Res, 2002), and to assess the validity of the high RBE values of 29 kV x-rays that had been reported. The experiments with the two types of x-rays were performed simultaneously and shared the same controls. The transformation yields with both radiation qualities were fitted to the linear-quadratic dependence on absorbed dose, and a corresponding analysis was performed for the data earlier obtained by Frankenberg et al. The transformation yields in the present study exceed those in the earlier investigation substantially, and it appears that the difference reflects inadequate feeding conditions of the cell cultures in the early experiments. The standard error bands of the dose response curves are derived and are seen to be considerably more narrow in the present results. The lowest dose of the 29 kV x rays was 1 Gy in both studies, and at this dose the RBE vs. the conventional x rays has now been found to be 2 with a 95% confidence interval of 1.4-2.6. The previous result was about 3.2, but the 95% confidence is very broad for these data. The estimated limit at low doses is 3.4 in the present experiments with a confidence interval that extends from less than 2 to large values. PMID- 14551784 TI - p53, Ki67 and cyclin D1 as prognosticators of lymph node metastases in laryngeal carcinoma. AB - The prognosis in patients suffering from head and neck squamous cell carcinomas depends on many factors. However, regional lymph node metastases are the most important parameter in determining the cure and survival of patients with head and neck cancers. The evaluation of cancer cell biology enables differentiation of their proliferation and tendency of metastases. Immunohistochemical examinations complement the well-established routine histological examination. The aim of this study was to evaluate the prognostic importance of the level of immunoproliferating proteins such as cyclin D1, nuclear antigen Ki67 and suppressor gene p53 for regional lymph node metastases in laryngeal carcinoma. The research was carried out on 73 patients treated for squamous cancer of the larynx in the Department of Otolaryngology University School of Medical Sciences in Poznan in the years 1994-1999. The group was comprised of 4 female and 69 male patients. Their ages ranged from 37 to 79 years, with a mean of 59 years. Clinical data included sex, age, localization and local and regional extent of the tumor, presence or lack of distant metastases, treatment, histological examination as well as immunohistochemical evaluation of suppressor gene p53, proliferative proteins Ki67 and cyclin D1. No statistically significant correlation was found between staining intensity of suppressor gene p53, cyclin D1 and the degree of local advancement (T). There was no correlation between the level of immunoproliferative markers and regional lymph node metastases. Statistically significant correlation was found between T stage and staining for Ki67 (P=0.017) as well as between cyclin D1 level and Ki67 (P<0.05). In conclusion, (1) no significant correlation was found between Ki67 and cyclin D1, p53 and TNM classification; (2) lack of correlation was confirmed between N+, p53, Ki67, cyclin D1 and Jacobsson classification; (3) the degree of histological grading correlated, however, with Jacobsson classification and cyclin D1 expression. PMID- 14551785 TI - Asymptomatic bilateral mixed-type laryngocele and laryngeal carcinoma. AB - A laryngocele is an abnormal dilatation of the laryngeal saccule. The relationship between layngoceles and laryngeal carcinoma is still debated. We present the case of a 45-year-old male patient with bilateral asymptomatic laryngocele and laryngeal squamous cell carcinoma. We discuss the possible etiopathogenetic influence of increased intralaryngeal pressure with preoperative CT and explorative findings, and the current literature is reviewed. PMID- 14551786 TI - Basic fibroblast growth factor expression in recurrent versus non-recurrent nasal polyposis. AB - Various growth factors are expressed in nasal polyps, and some of these have been suggested to play a role in polyp formation. A potential relation between growth factor expression and polyp recurrence, however, is undetermined. Basic fibroblast growth factor (bFGF) is expressed in mononuclear cells, as well as in endothelial and epithelial surface and gland cells of nasal polyps. To determine whether bFGF may play a role in the recurrence of nasal polyps, the present study aimed at a comparison of bFGF expression in recurrent versus non-recurrent polyps. Further, the expression in polyps from asthmatic patients was compared with that from non-asthmatics. Thirty patients with newly diagnosed nasal polyposis were included. Polypectomy was performed at entry to the long-term follow-up study. Fifteen patients only had one polypectomy (no recurrence group, with a median observation time of 81 months). Fifteen patients had a median of 6.4 polypectomies (multiple recurrence group, with a median observation time of 108 months). Five of nine patients with asthma belonged to the non-recurrence group and four to the recurrence group. The polyp from the entrance polypectomy was examined for expression of bFGF by immunohistochemistry, using a polyclonal antibody. A masked semi-quantification of staining intensity was performed in recurrent versus non-recurrent polyps, as well as in asthmatics versus non asthmatics. bFGF expression was seen as varying staining of the polyp surface and gland epithelium, as well as of some mononuclear cells and some fibroblast-like cell profiles in the polyp stroma. Vascular endothelium was labeled occasionally. Semi-quantification of the staining intensity showed no significant differences between recurrent and non-recurrent polyps, or between asthmatics and non asthmatics. We conclude that the level of immunohistochemical expression of bFGF in recurrent and non-recurrent nasal polyposis is equivalent. Thus, the level of bFGF expression in the primary polyp can not predict a subsequent recurrence. The expression of bFGF is not up-regulated in patients with asthma. Further studies are needed to determine a potential role of bFGF in nasal polyposis, with special reference to different stages of polyp formation and growth. PMID- 14551787 TI - Laser myringotomy in otitis media with effusion: long-term follow-up. AB - Otitis media with effusion is a leading cause of conductive hearing loss in children. Myringotomy and insertion of tympanostomy tubes is the accepted form of treatment. Recently, several studies utilizing laser myringotomy have been published, but few of them present late results. The objective of this study was to compare late results of the treatment with laser and classical myringotomy. A clinical effectiveness trial was conducted in three groups of children: (1) 37 children treated with laser myringotomy (ML), (2) 29 children treated with laser myringotomy and the insertion of tympanostomy tubes (ML+V) and (3) 43 children treated with classical myringotomy and the insertion of tympanostomy tubes (MC+V). All types of surgery were performed under general anesthesia because adenoidectomy and/or tonsillectomy was done at the same time. The results of treatment were assessed on the basis of the otoscopic examination (recurrences of effusion, condition of the tympanic membrane, and audiological examination (pure tone audiometry, tympanometry and DPOAE). The minimum follow-up period was 1 year. The recurrence rate was lowest in the ML+V (11%) group, and highest in the ML group (36%). The difference between ML+V and MC+V was not significant. Permanent changes in the tympanic membrane were observed in 8% of the ears after ML, 19% after ML+V and 31% after MC+V. The difference was significant between the ML and MC+V groups. PTA was significantly higher in the MC+V group than in the control group of otologically healthy children. Mean amplitudes of DPOAE, measured in treated children with normal tympanometry results, were significantly lower than in the control group, but within the normal range. The use of CO(2) laser during myringotomy has no negative effect on the function of the cochlea. Healing of the tympanic membrane after laser myringotomy was uneventful with a low percentage of permanent sequelae. PMID- 14551788 TI - Individual prefabricated titanium implants and titanium mesh in skull base reconstructive surgery. A report of cases. AB - Titanium implants can be shaped by traditional hand forming, press shaping, modular construction by welding, construction on full-size models shaped from CT coordinates and, most recently, by computer-assisted design and computer-assisted manufacturing (CAD/CAM) that consist in the direct prefabrication of individual implants by milling them out of a solid block of titanium. The aim of our study was to present a set of preliminary cases of an ongoing program of reconstructive procedures of the skull base using titanium implants. The subjects underwent ablative procedures of the skull base with reconstruction either by titanium mesh or individual prefabricated CAD/CAM implants. Six patients have been operated on successfully since 2000: two received prefabricated CAD/CAM titanium plates and four others underwent reconstruction with titanium mesh. The stability of CAD/CAM plates is superior to that of mesh, thus it is more useful in reconstructing large lesions of the frontal skull base and the temporal and occipital bones. Titanium mesh was successfully used for defects smaller than 100 cm(2) or where selected viscerocranial defects are complicated in design and less reproducible by CAD/CAM. The intraoperative design, shaping and adjustment characteristic of titanium mesh can be dispensed with when CAD/CAM implants are used. The 3-D data set used in the CAD/CAM process also operates in the navigated simulation and planning of the ablation contours, the latter being of great assistance in establishing the optimal future defect. As a disadvantage, CAD/CAM technology is more expensive than titanium mesh, and the process is time-consuming as it is carried out in advance of surgery. PMID- 14551789 TI - An audit of pharyngeal pouch surgery using endoscopic stapling. The patient's viewpoint. AB - We report an audit designed to assess patient satisfaction resulting from pharyngeal pouch surgery using an endoscopic stapler. A personal series of 16 patients all operated on by the senior author over a 3-year period is reported. Information was gathered using a retrospective telephone questionnaire to establish pre- and postoperative symptoms, complications of surgery and patient satisfaction. This showed endoscopic pharyngeal pouch surgery to be successful in the majority of cases. Eighty-seven percent of patients felt better as a result of surgery. Seventy-five percent had no symptoms postoperatively. This series shows that surgery on pharyngeal pouches results in significant improvement in patient symptoms with minimal morbidity and mortality. PMID- 14551790 TI - Multisystem Langerhans' cell histiocytosis (Hand-Schuller-Christian disease) in an adult: a case report and review of the literature. AB - Langerhans' cell histiocytosis (LCH) is a rare and enigmatic clonal disorder that affects mainly children. It is characterized by single or multiple granulomatous mass lesions composed of cells with the Langerhans' cell phenotype. Clinical presentation and behavior are heterogeneous and can range from a solitary lytic bone lesion (i.e., eosinophilic granuloma) with a favorable course to a fatal disseminated leukaemia-like form, with a wide spectrum of intermediate clinical presentations between these two extremes. Although LCH typically involves the bone, lesions can be found in almost all organs. We are reporting the case of a multisystem LCH in a 47-year-old patient who presented with a panhypopituitarism and diabetes insipidus, and who, 5 years later, developed mandibular, mastoid and femoral lesions. The final diagnosis of LCH was made on mandibular biopsy. PMID- 14551791 TI - Collection of nasal secretions for immunological analysis. AB - The biochemical analysis of nasal secretions has become essential in the study of nasal or sinus diseases and the monitoring of medical and surgical treatment. The nasal fluid greatly reflects the inflammatory activity within the nasal mucosa. This paper discusses techniques for nasal fluid collection described before and proposes a new approach for the collection and calculation of nasal secretions based on sinus packs. The method is non-invasive, well standardized and reproducible and therefore may serve as a valid tool for future investigations. PMID- 14551792 TI - Plexiform ameloblastoma presenting as a sinonasal tumor. AB - Ameloblastomas are the most frequent odontogenic tumours, accounting for 1% of all tumours of the maxilla and mandible. Sinonasal ameloblastomas are most common between the ages of 55 and 65, and mandibular ameloblastomas between 40 and 50. Incidence is higher in males than in females, and there are no differences between races. These locally aggressive tumours originate in the mandible in 80% of cases and in the maxilla in 15-20%. We report an unusual primary nasosinusal ameloblastoma presented in a 68-year-old male. The tumour was completely resected by (para)lateral rhinotomy and treated with postoperative radiotherapy. Histological analysis demonstrated a plexiform ameloblastoma. The patient remains well without disease after 50 months of postoperative follow-up. PMID- 14551793 TI - T1-T2 NO oropharyngeal cancers treated with surgery alone. A GETTEC study. AB - The aim of this study is to show that surgical treatment of early-stage squamous cell carcinomas of the oropharynx gives identical, if not better, oncological results than the classic radiotherapy treatment in terms of locoregional control and survival. Fifty-three patients (32 T1, 21 T2, all N0) were operated on during the years 1995-2000. Surgical treatment consisted in a resection by the transoral approach in 43 patients (81.13%); ten patients (18.87%) benefited from a pharyngectomy with (seven) or without (three) mandibular resection. A level I to V selective neck dissection was performed on 35 patients, and 5 patients underwent a level II to V selective neck dissection. The 1-, 3- and 5-year overall survival rates were 100, 94.6 and 73%, respectively. There was no significant difference concerning the tumor stage ( P=0.69), the initial localization ( P=0.64), the macroscopic aspect ( P=0.65) and the management undertaken in the different centers ( P=0.19). The 5-year rate of specific survival was 100%. The 1-, 3- and 5-year locoregional control rates were 96.22, 92.45 and 88.68, respectively. The oncological occurrences observed were 2 persistent diseases, 5 local recurrences, 11 second primary cancers and 0 nodal recurrences. Seven local failures were observed, all of which were controlled after a second treatment. Eleven patients presented second primary cancers; three died, two are alive with an extension of this second localization, and six are alive and free of disease. The locoregional control provided by surgery alone on T1-T2 N0 oropharyngeal cancers is as good as radiotherapy. Moreover surgery alone makes it possible to spare patients the complications and aftereffects of radiotherapy. It also makes it possible during the recurrences to operate on patients in non-irradiated areas with lower morbidity and mortality. It is all the more beneficial since it will be possible to resort to radiotherapy after surgery if need be. PMID- 14551794 TI - Cavernous hemangioma of the left vocal cord. AB - Laryngeal hemangioma in adults is a very rare condition. We report the case of a 41-year-old man with hemangioma of the left vocal cord. The literature on this rare occurrence is reviewed and the significance of the case discussed. PMID- 14551795 TI - Maternal peripheral T helper 1-type and T helper 2-type immunity in women during the first trimester of twin pregnancy. AB - INTRODUCTION: This study investigated the T helper Th1:Th2 balance in twin pregnancies compared with singleton pregnancies during the first trimester. METHODS: Blood samples were taken from 24 women with a singleton pregnancy and 14 women with twin pregnancy at 8-9 weeks' gestation to examine the ratios of Th1:Th2 and serum human chorionic gonadotropin (hCG) and progesterone levels. RESULTS: The average ratio of Th1:Th2 in the twin pregnancies was significantly lower than that in singleton pregnancies (7.3+/-2.3 vs. 10.5+/-2.2, p<0.05). There were negative correlations between the Th1:Th2 ratio and serum hCG levels (mIU/ml) (Th1:Th2 ratio = 14.5-4.52 x 10(-5)xhCG, r(2)=0.41, p<0.05) and between the Th1:Th2 ratio and serum progesterone levels (ng/dl; Th1:Th2 ratio = 23.0-0.63 x progesterone, r(2)=0.36, p<0.05). CONCLUSION: Our findings show marked predominance of Th2 type cytokines occurring in twin pregnancies is related to the increase in trophoblasts during the first trimester. PMID- 14551796 TI - A primary retroperitoneal serous cystadenocarcinoma with clinically aggressive behavior. AB - CASE REPORT: We describe a 44-year-old woman with a primary retroperitoneal serous cystadenocarcinoma as the fourth report in the world literature. A contrast-enhanced computed tomographic scan of the abdomen demonstrated a mass with enhanced solid mural nodules and septa in the left retroperitoneum. Serum CA19-9 was elevated at 181 U/ml. The patient underwent a complete resection of the retroperitoneal tumor with a partial resection of the psoas major muscle and its fascia. Pathological examination showed a well-differentiated papillary serous cystadenocarcinoma of ovarian type and locoregional lymph node metastases. Seven months after surgery, the patient developed a pelvic recurrence, and underwent a total hysterectomy, a left salpingo-oophorectomy and a resection of the metastatic mesenteric mass. Two months after the second surgery, serum CA19-9 and CA125 increased to 1,114 U/ml and 218 U/ml, respectively. A solitary liver metastasis was confirmed on a computed tomographic scan. The patient is now being treated with weekly docetaxel and carboplatin. CONCLUSION: The present case illustrates the clinically aggressive nature of a primary retroperitoneal serous cystadenocarcinoma. PMID- 14551797 TI - Synchronous cancer and metastatic melanoma of the breasts. AB - CASE REPORT: We present a synchronous primary cancer in the right breast and the metastasis of a malignant melanoma in the left breast. Both lesions were excised and the right breast was irradiated. DISCUSSION: Problems associated with the simultaneous occurrence of both diseases are briefly discussed. PMID- 14551798 TI - Passive transfer of purified IgG from patients with amyotrophic lateral sclerosis to mice results in degeneration of motor neurons accompanied by Ca2+ enhancement. AB - It has been reported that immunoglobulins (IgG) in sera of patients with amyotrophic lateral sclerosis (ALS) kill cultured motoneurones (MN), but whether they also cause MN degeneration in vivo is unclear. To test this, protein-A affinity purified and dialysed IgGs were prepared from sera of 44 ALS patients without paraproteinemias, 20 healthy controls and 15 disease controls. Control and ALS-IgGs were injected intraperitoneally into groups of mice for 5 consecutive days and examined at day 8. IgG was localised immunocytochemically and spinal MN were characterised histologically and ultrastructurally and by comparative counts of Ca(2+) containing organelles revealed with oxylate pyroantimonate histochemistry. ELISA revealed no differences in IgG concentration between ALS patients and control subjects. Immunocytochemistry showed IgG was present in MN of mice injected with control or ALS-IgG, but densitometry showed immunostaining in MN was stronger in mice injected with ALS-IgG. Compared to MN of non-injected mice, control-IgG-treated mice showed near normal MN morphology and numbers of Ca(2+)-containing organelles. Disease control IgGs evoked negligible or minor morphological changes according to disease, but normal numbers of Ca(2+) containing organelles. Ultrastructurally, about 70% of ALS derived IgGs induced a population of MN with electron lucent cytoplasm, distended Golgi, disrupted Nissl and mitochondria (i.e., necrosis). However 30% of ALS-IgGs additionally induced electron-dense degeneration in 40% of the MN. These MN exhibited shrinkage, condensed nuclear chromatin and ill-defined nuclear membranes and resembled preliminary stages of apoptosis. We conclude that passive transfer of ALS-derived, but not control IgGs, does result in MN degeneration in the recipient mice. This appears to be associated with abnormal calcium homeostasis, but the exact target of ALS-IgG remains conjectural, and the possibilities are discussed. PMID- 14551799 TI - PRESS-based proton single-voxel spectroscopy and spectroscopic imaging with very short echo times using asymmetric RF pulses. AB - Modified point-resolved spectroscopy (PRESS) sequences for single voxel spectroscopy (MRS) and spectroscopic imaging (SI) with very short echo time ( T(E)) are described using asymmetric radio-frequency (RF) pulses as well as an optimized design and timing of the PRESS sequence. The proposed sequences were implemented on a standard 4.7 T imaging system yielding a T(E) of 6.0 ms only. Simulations and experimental data measured on phantoms and the rat brain in vivo are presented for MRS and SI showing a high signal-to-noise ratio and hardly any phase distortions caused by J-coupling. PMID- 14551801 TI - Silent invasion: Imanishi's primatology and cultural bias in science. PMID- 14551802 TI - Honor established writings. PMID- 14551803 TI - [Root growth after wisdom tooth transposition. Case documentation with proposal for a radiologic method to quantify growth]. AB - OBJECTIVES: To introduce a method for assessment of root growth on panoramic radiographs exemplified by a case report on an autotransplanted lower third molar. MATERIAL AND METHODS: A lower left third molar (T) with incomplete root formation was transplanted for replacement of the lower left first molar in an 18 year-old woman. T was monitored clinically and, together with its neighboring lower second molar (37) with constant root length, also radiographically by means of two panoramic radiographs (OPGs) produced 12 days and 20 months postoperatively. A method for calculation of the root length as a multiple of the crown length is introduced, using accurately reproducible landmarks defining the coronal and apical endpoints of the examined tooth in all OPGs of the series. This method minimizes error due to different magnifications within the set of radiographs. RESULTS: Using the method introduced, the root length of the constant tooth 37 varied at 2.7% within the set of OPGs, whereas it revealed a 5.6% variation when the evaluation was based on direct measurement. Based on the described method, T revealed a postoperative root growth of approximately one third of its final length and showed clinically no pathological findings during the observation period. CONCLUSIONS: Our results indicate that with the described method root growth assessment on panoramic radiographs is more accurate than with direct measurement. PMID- 14551804 TI - [In vivo study of degradation of poly-(D,L-) lactide and poly-(L-lactide-co glycolide) osteosynthesis material]. AB - AIMS: Comparison of the degradation of poly(D,L)lactide (Resorb X) or poly(lactide-co-glycolide) (LactoSorb) in vivo. MATERIAL AND METHODS: LactoSorb and Resorb X osteosynthesis plates were fixed at the lateral aspect of the femora of 26 Chinchilla rabbits using the respective osteosynthesis screws. After intraperitoneal injection of fluorochromes the screw plate bone blocks were resected after 1, 6, 12, 14, 16, 21, 26 months and radiologic, histologic as well as fluorescence microscopic examinations were carried out. RESULTS: Newly formed bone was detectable above and beneath the polymers 1 month after the implantation. The implants were totally covered by newly formed bone after 6 months. While the LactoSorb screws were found to be as birefringent as after 1 month, in the Resorb X screws a continuous resorption by phagocytizing marrow cells starting from the periphery was detectable. Resorb X was totally resorbed in histologic slides 12 months after implantation, while total resorption of LactoSorb lasted 14 months; both polymers were replaced by marrow cells. Bone remodeling was not finished 26 months after implantation in both polymers. CONCLUSION: Resorption of Resorb X was finished earlier than the resorption of LactoSorb. Both materials were found by fluorescence microscope to be completely resorbed after 12 or 14 months, but bone remodeling of the screw holes was not yet finished 26 months after implantation. PMID- 14551805 TI - [3-dimensional imaging possibilities of thhe mid-face area using digital volume tomography based on a daver study of angle stable osteosynthesis]. AB - BACKGROUND: Multidirectional angularly stable interlocking plate systems are now available for the surgical treatment of the midface. After first experiences in cases of mandibular fractures, application to the facial skeleton was investigated using a cadaver study. Furthermore, three-dimensional imaging by means of the digital volume tomograph NewTom DVT 9000 after reduction of orbitozygomatical fractures was performed and evaluated. MATERIAL AND METHODS: After artificial osteotomy and reduction of both zygomatical complexes, osteosynthesis of the left side was performed with three 4-hole miniplates (2.0). On the right side, three angular stable 2-hole plates (2.3) were used. Thereafter, a three-dimensional data set was generated using the NewTom DVT 9000. After DICOM-import in eFilm reconstructions were evaluated by six examiners regarding defined criteria. RESULTS: After adaptation and fixation of the angular stable interlocking plate system without complications, manual checking revealed that the primary stability did not seem inferior to the other side. Defined criteria were sufficiently evaluable, even close to the osteosynthetic material, using reconstructions of the digital volume tomography data set. CONCLUSION: Surgical treatment of midfacial fractures using a multidirectional angularly stable interlocking plate system seems promising. The NewTom DVT 9000 proved to be suitable to visualize even fine osseous structures of the midface. PMID- 14551806 TI - [Evidence of osteocalcin expression in osteoblast cells of mandibular origin growing on biomaterials with RT-PCR and SDS-PAGE/Western blotting]. AB - A new approach to addressing difficult tissue reconstructive or replacement problems in the oral cavity is to engineer new tissue by using selective cell transplantation on polymer scaffolds. The current study characterized the osteoblastic nature of adherent mandibular cells on biomaterials, which could have a potential use as scaffolds for tissue engineering strategies. Cells of mandibular origin from one patient were cultivated on three different biomaterials (PepGen P-15 trade mark, Frios Algipore, and OsteoGraf/LD-700) for 7 and 14 days and osteocalcin expression was demonstrated by RT-PCR and SDS PAGE/Western blotting. In order to explicitly characterize only the adherent cells on the biomaterials, we first separated the biomaterials with adherent cells from the culture plate before trypsinization. We could demonstrate that cell growth of adherent mandibular osteoblast-like cells was significantly higher on biomaterials with an organic component (PepGen P-15 trade mark ) in comparison to Frios Algipore and OsteoGraf/LD-700, respectively. In conclusion, only the explicit study of adherent cells at the gene and protein levels gives information about the osteoconductivity of biomaterials. PMID- 14551807 TI - [Adenomatoid odontogenic tumor in calcifying odontogenic cyst]. AB - BACKGROUND: Adenomatoid odontogenic tumors (AOT) and calcifying odontogenic cysts (COC, Gorlin's cyst) are rare benign specificities of odontogenic tumors. Most odontogenic tumors are lesions of embryonic odontogenic tissue. Radiographically, odontogenic tumors may mimic an odontogenic follicular cyst. CASE REPORT: This paper describes the case of an 11-year-old female patient with a remarkable combination of these two entities. The lesions were found at the site of a retained first premolar and a persistent deciduous tooth 84 of the right lower jaw. Without clinical symptoms the tumor had expanded the vestibular cortical bone of the premolar region. Radiographically, the lesion appeared as a typical follicular cyst. During the operation, excochleation of the cyst was performed and the premolar was extracted. Finally, the epithelium of the cyst was treated by curettage. The histologic specimen revealed the diagnosis of an adenomatoid odontogenic tumor (AOT) in a calcifying odontogenic cyst (COC). DISCUSSION: The described case shows that symptoms of odontogenic tumors are usually nonspecific. For that reason one should always take a biopsy for histological investigation when patients present atypical cysts of the jaw. In rare cases even malignant odontogenic neoplasms could occur. PMID- 14551808 TI - [Condylar reconstruction after resection of an intracapsular stomach carcinoma metastasis]. AB - BACKGROUND: The mandible is a very uncommon place for a metastasis of a gastric carcinoma. Normally the area of the temporomandibular joint (TMJ) remains unaffected. The separate vascularization is discussed as one reason among others. Primary reconstruction after resection of the condyle is often problematic because an early onset of adjuvant systemic therapy is required. In this case, the insertion of a Quinn joint prosthesis is presented after resection of a TMJ metastasis. CASE: We report a hematogenic metastatic gastric adenocarcinoma in a 51-year old male who initially presented with increasing disclusion in the left molar region. Suspecting a metastatic adenocarcinoma of the TMJ, a condylectomy with immediate replacement by a total joint prosthesis was performed via a preauricular approach. Corresponding to the clinically and radiologically suspected diagnosis, the decalcified histological specimen presented as a metastatic gastric adenocarcinoma within the intracapsular region. RESULTS: The healing period of the implanted modified Quinn prosthesis was fast and uncomplicated after resection of this, to our knowledge, first documented metastatic gastric adenocarcinoma of the intracapsular region. After early restoration of joint function and patient satisfaction, the required radiochemotherapy of further unresectable bony metastases could be started in time. DISCUSSION: This example of an extremely rare case of a metastasis shows that such a total joint prosthesis appears to be a very good alternative to extended autogenous reconstruction or an unsatisfactory primary resection. Due to the mating of the spherical condylar head and glenoid fossa, the modified Quinn prosthesis is very suitable for total joint replacement after extended resection or in multiply preoperated cases. PMID- 14551810 TI - Kinetics and mechanism of iron release from the bacterial ferric binding protein nFbp: exogenous anion influence and comparison with mammalian transferrin. AB - Ferric binding protein, Fbp, serves an essential biological function in shuttling naked (hydrated) Fe(3+) across the periplasmic space of many Gram-negative bacteria. In this process, iron must be released at the cytoplasmic membrane to a permease. How iron is released from Fbp has yet to be resolved. Consequently, understanding the dynamics of iron release from Fbp is of both biological and chemical interest. Fbp requires an exogenous anion, e.g. phosphate when isolated from cell lysates, for tight iron sequestration. To address the role of exogenous anion identity and lability on Fe(aq)(3+) dissociation from Fbp, the kinetics of PO(4)(3-) exchange in Fe(3+) nFbp(PO(4)) ( nFbp=recombinant Fbp from Neisseria meningitidis) were investigated by dynamic (31)P NMR and the kinetics of Fe(3+) dissociation from Fe(3+) nFbp(X) (X=PO(4)(3-), citrate anion) were investigated by stopped-flow pH-jump measurements. We justify the use of non-physiological low pH conditions because a high [H(+)] will drive the Fe(aq)(3+) dissociation reaction to completion without using competing chelators, whose presence may complicate or influence the dissociation mechanism. For perspective, these studies of nFbp (which has been referred to as a bacterial transferrin) are compared to new and previously published kinetic and thermodynamic data for mammalian transferrin. Significantly, we address the lability of the Fe(3+) coordination shell in nFbp, Fe(3+) nFbp(X) (X=PO(4)(3-), citrate), with respect to exogenous anion (X(n-)) exchange and dissociation, and ultimately complete dissociation of the protein to yield naked (hydrated) Fe(aq)(3+). These findings are a first step in understanding the process of iron donation to the bacterial permease for transport across the cytoplasmic membrane. PMID- 14551812 TI - Influenza B virus NS2, a nuclear export protein, directly associates with the viral ribonucleoprotein complex. AB - In Influenza A virus and Influenza B virus, the NS2 protein (nuclear export protein) has been proposed to mediate the nucleocytoplasmic trafficking of viral ribonucleoprotein (vRNP) by forming NS2-vRNP complexes. While the binding interactions of NS2 in these complexes have been well characterized for Influenza A virus, much less is known about Influenza B virus NS2 (B/NS2). In this report, we developed a specific antiserum against B/NS2 protein and demonstrated that B/NS2 was synthesized late in infection and packaged into virions after nucleocytoplasmic transport. Fractionation of detergent-disrupted virions in several conditions showed that B/NS2 remained associated with vRNP after separation of matrix protein M1 from vRNP, whereas Influenza A virus NS2 (A/NS2) was easily separated from vRNP and remained associated with M1, in accord with previous findings that A/NS2 associates with vRNP only through its binding of encapsidated M1. The results indicated that complex formation among vRNP, M1 and NS2 of Influenza B virus was different from that of Influenza A virus, and that B/NS2 associated with vRNP in the absence and presence of M1. PMID- 14551813 TI - Detection of antibody for the serodiagnosis of hantavirus infection in different rodent species. AB - Peroxidase-labeled staphylococcal protein A, streptococcal protein G, and antibodies directed against Mus musculus (mouse), Rattus norvegicus (rat), Mesocretus auratus (hamster), and Peromyscus leucopus were examined for their reactivity with immunoglobulin G (IgG) from various rodent species. The purpose of this study was to identify the optimal secondary antibodies or reagents for specific serodiagnosis of hantavirus infection in various rodent species. Using ELISA, a total of 65 sera from 29 rodent species of the family Muridae and one serum sample from family Octodontidae were compared for IgG reactivity with the six different reagents. The results demonstrate that the reactivities of the secondary antibodies and reagents to the sera varied, even among sera from rodents of the same genus. Hantavirus-specific antibody ELISA revealed that hantavirus-infected rodent sera obtained from M. musculus, R. norvegicus, Apodemus agrarius, A. peninsulae, and Bandicota indica bound to the six different conjugates in a similar pattern as that detected in IgG ELISA. These results indicate that the applicability of secondary antibodies and protein A and G should be carefully evaluated before use for serodiagnosis in different rodent species. PMID- 14551811 TI - Synthesis, structure analysis, solution chemistry, and in vitro insulinomimetic activity of novel oxovanadium(IV) complexes with tripodal ligands containing an imidazole group derived from amino acids. AB - Structures, chemical properties, and in vitro insulinomimetic activities of new vanadyl [oxovanadium(IV), VO(2+)] complexes with five tripodal ligands containing an imidazole functionality were examined. The ligands, N-(carboxymethyl)- N-(4 imidazolylmethyl)amino acids, contain glycine, ( S)- and ( R)-alanine, and ( S)- and ( R)-leucine residues. The molecular structures of the latter four alanine- and leucine-containing complexes were determined by X-ray analysis. The coordination geometry around each vanadium center was octahedral, where an imino nitrogen occupied the apical site and two carboxylate oxygens, an imidazole nitrogen, and a water molecule coordinated in the equatorial plane. The spectroscopic properties of the complexes were characterized by means of IR, electronic absorption, and CD spectra. Acid dissociation constants (p K(a)) and protonation sites of the ligands were determined by a combination of potentiometric titrations and (1)H NMR spectra. The potentiometric study demonstrated that stability constants (log beta) were not so different among the present complexes (14.0-14.9) and a species of molecular complex with a 1:1 metal:ligand ratio existed predominantly at physiological pH 7.4. EPR parameters indicated that the species at pH 7.4 had an octahedral structure similar to the complex in the solid state. On the other hand, an EPR study in phosphate buffer (pH 7.4) suggested that inorganic phosphate coordinated to the vanadium center instead of the imidazole group in the presence of excess phosphate ion. Cyclic voltammograms in the phosphate buffer showed chemically reversible oxidation waves, whereas irreversible oxidation waves were observed in non-coordinating HEPES buffer. Moreover, the oxidation potential of each complex in phosphate buffer was more positive than that in HEPES buffer. Partition coefficients of the present complexes in a n-octanol/saline system were very low, probably due to hydrophilicity of the imidazole group. The in vitro insulinomimetic activities were estimated on the basis of the ability of the complexes to inhibit epinephrine-stimulated free fatty acid release from isolated rat adipocytes. The achiral glycine-derivative complex exhibited the highest insulinomimetic activity, which was higher than that of VOSO(4) as a positive control. Putting our previous observations together, it was found that the vanadyl complexes with tetradentate amino acid derivatives having no alkyl side chain tend to have high in vitro insulinomimetic activity. PMID- 14551814 TI - Genistein, a protein tyrosine kinase inhibitor, suppresses the fusogenicity of Moloney murine leukemia virus envelope protein in XC cells. AB - XC cells are highly susceptible to syncytium formation by infection of ecotropic murine leukemia viruses (MLVs) and by expression of their envelope protein (Env). By this property, XC cells are widely used to determine titers of ecotropic MLVs. Number of plaques resulted from the syncytium formation in XC cells by ecotropic MLV infection is corresponding to number of the viral particles. XC cells had been established from a v-src-induced rat tumor. It has been reported that transformed cells are more sensitive to Mo-MLV-induced syncytium formation than non-transformed cells. To assess whether the transformation by v-src oncogene in XC cells is involved in the high sensitivity to ecotropic MLV-induced syncytium formation, XC cells were treated with genistein, a protein tyrosine kinase inhibitor. Genistein suppressed the syncytium formation between XC cells and ecotropic Env-expressing 293T cells. This result indicates that protein tyrosine kinase activity is associated with the high sensitivity of XC cells to ecotropic Env-induced syncytium formation. PMID- 14551815 TI - Characterization of the 3'-terminal nucleotide sequence of two Korean isolates of Daphne virus S support its placement as a distinct species of the genus Carlavirus. AB - This study determined the 3'-terminal nucleotide sequences of two Korean isolates of Daphne virus S (DVS), a tentative member of the genus Carlavirus, causing leaf distortion and chlorotic spot disease symptoms in daphne plants. The 3'-terminal 1,465 nucleotide sequences of the two isolates contained two open reading frames coding for proteins of 36 kDa viral coat protein (CP) and 12 kDa from the 5'-3' end, which is a typical genome structure of the 3'-terminal region of carlaviruses. Both DVS isolates were 98.1% and 93.6% amino acid identical in the CP and 12 kDa, respectively. The CP gene of DVS shares 25.2-55.2% and 42.9-56.1% similarities with that of 19 other carlaviruses at the amino acid and nucleotide levels, respectively. The 3'-proximal 12 kDa gene of DVS shares 20.2-57.8% amino acid identities with that of 18 other members of the genus. The 3' noncoding region of DVS consists of 73 nucleotides with long excluding poly A tract, and shares 69.1-77.1% identities to the known carlaviruses. In the phylogenetic analyses of the two proteins, DVS was closely related to Helenium virus S and Chrysanthemum virus B. This is the first sequence information for the DVS, and further confirms the classification of DVS as a distinct member of the genus Carlavirus. PMID- 14551816 TI - Variability and transmission by Aphis glycines of North American and Asian Soybean mosaic virus isolates. AB - The variability of North American and Asian strains and isolates of Soybean mosaic virus was investigated. First, polymerase chain reaction (PCR) products representing the coat protein (CP)-coding regions of 38 SMVs were analyzed for restriction fragment length polymorphisms (RFLP). Second, the nucleotide and predicted amino acid sequence variability of the P1-coding region of 18 SMVs and the helper component/protease (HC/Pro) and CP-coding regions of 25 SMVs were assessed. The CP nucleotide and predicted amino acid sequences were the most similar and predicted phylogenetic relationships similar to those obtained from RFLP analysis. Neither RFLP nor sequence analyses of the CP-coding regions grouped the SMVs by geographical origin. The P1 and HC/Pro sequences were more variable and separated the North American and Asian SMV isolates into two groups similar to previously reported differences in pathogenic diversity of the two sets of SMV isolates. The P1 region was the most informative of the three regions analyzed. To assess the biological relevance of the sequence differences in the HC/Pro and CP coding regions, the transmissibility of 14 SMV isolates by Aphis glycines was tested. All field isolates of SMV were transmitted efficiently by A. glycines, but the laboratory isolates analyzed were transmitted poorly. The amino acid sequences from most, but not all, of the poorly transmitted isolates contained mutations in the aphid transmission-associated DAG and/or KLSC amino acid sequence motifs of CP and HC/Pro, respectively. PMID- 14551817 TI - The complete nucleotide sequence of the genome RNA of Lily symptomless virus and its comparison with that of other carlaviruses. AB - The complete genomic nucleotide sequence and genome structure of Lily symptomless virus (LSV), a lily-infecting carlavirus, have been obtained. The genome of the Korean strain of LSV, LSV-Kr, was 8394 nucleotides long and contained six open reading frames (ORFs) coding for proteins of Mr 220 kDa (1948 aa), 25 kDa (228 aa), 12 kDa (106 aa), 7 kDa (64 aa), 32 kDa (291 aa) and 16 kDa (140 aa) from the 5' to 3' end, respectively, which is typical of carlaviruses. Genetic heterogeneity was observed in the ORF1 gene. A total of 221 of 5,847 nucleotides (nt) were heterologous in the ORF1 of replicase; 162 nt portions were silent and 59 nt resulted in amino acid changes. This heterogeneity indicates that the LSV infecting lily plants contained a genetically heterogeneous population of LSV (quasispecies). Overall similarities to those of other carlaviruses for the six ORFs of LSV were from 76.1% to 31.6% and from 87.3% to 13.7%, at nucleotide and amino acid levels, respectively. The ORF1 replicase gene of LSV shares 40.9% to 56.8% and 48.9% and 58.6% identities with that of 5 other carlaviruses at the amino acid and nucleotide levels, respectively. LSV was closest to Blueberry scorch virus (BlScV) in this ORF, among the carlaviruses for which sequence information is available. The three triple gene blocks (ORF2-4), ORF5 (coat protein) and 3'-proximal 16 kDa ORF6 genes were further analyzed, and phylogenetic trees for the coding regions indicate that the LSV was the most closely related to Kalanchoe latent virus and BlScV. This is the first report of the complete nucleotide sequence and genome structure of LSV. PMID- 14551818 TI - Sequence diversity of South Pacific isolates of Taro bacilliform virus and the development of a PCR-based diagnostic test. AB - We have analysed the sequence variability in the putative reverse transcriptase (RT)/ribonuclease H (RNaseH) and the C-terminal coat protein (CP)-coding regions from Taro bacilliform virus (TaBV) isolates collected throughout the Pacific Islands. When the RT/RNaseH-coding region of 22 TaBV isolates from Fiji, French Polynesia, New Caledonia, Papua New Guinea (PNG), Samoa, Solomon Islands and Vanuatu was examined, maximum variability at the nucleotide and amino acid level was 22.9% and 13.6%, respectively. Within the CP-coding region of 13 TaBV isolates from Fiji, New Caledonia, PNG, Samoa and the Solomon Islands, maximum variability at the nucleotide and amino acid level was 30.7% and 19.5%, respectively. Phylogenetic analysis showed that TaBV isolates from the Solomon Islands showed greatest variability while those from New Caledonia and PNG showed least variability. Based on the sequences of the TaBV RT/RNaseH-coding region, we have developed a PCR-based diagnostic test that specifically detects all known TaBV isolates. Preliminary indexing has revealed that TaBV is widespread throughout Pacific Island countries. A sequence showing approximately 50% nucleotide identity to TaBV in the RT/RNaseH-coding region was also detected in all taro samples tested. The possibility that this may represent either an integrated sequence or the genome of an additional badnavirus infecting taro is discussed. PMID- 14551819 TI - Cotton leaf curl disease is associated with multiple monopartite begomoviruses supported by single DNA beta. AB - For bipartite begomoviruses (family Geminiviridae) trans-replication of the DNA B component by the DNA A-encoded replication-associated protein (Rep) is achieved by virtue of a shared sequence, the "common region", which contains repeated motifs (iterons) which are sequence-specific Rep binding sites and form part of the origin of replication. Recently cotton leaf curl disease (CLCuD), a major constraint to cotton production on the Indian subcontinent, has been shown to be caused by a monopartite begomovirus ( Cotton leaf curl Multan virus [CLCuMV]) and a novel single-stranded DNA satellite molecule termed CLCuD DNA beta. The satellite molecule is trans-replicated by CLCuMV but does not possess the iteron sequences of this virus. We have investigated the ability of CLCuD DNA beta to interact with three further clones of monopartite begomoviruses, isolated from cotton, that have distinct Rep binding specificities. All three cloned viruses were capable of trans-replicating the satellite molecule and inducing CLCuD symptoms in cotton, indicating that the interaction between begomovirus and DNA beta is relaxed in comparison to the interaction between DNA A and DNA B components. Field surveys across all the cotton growing regions of Pakistan indicate that dual and multiple infections are the norm for CLCuD with no evidence of synergism. Despite the diversity of begomoviruses associated with CLCuD, only a single class of DNA beta has been detected, suggesting that this satellite has the capacity to be recruited by unrelated begomoviruses. PMID- 14551820 TI - Multiple heparin binding domains of respiratory syncytial virus G mediate binding to mammalian cells. AB - Respiratory syncytial virus (RSV) G glycoprotein mediates cell attachment through surface glycosaminoglycans (GAGs). Feldman et al. [10] suggested that specific basic amino acids in residues 184-198 of G defined a critical heparin binding domain (HBD). To further define the G HBD we made a series of truncated G proteins expressed in Escherichia coli. G88 (G residues 143-231), bound to HEp-2 cells in a dose dependent manner and binding was inhibited >99% with heparin. Cell binding of G88 was unaltered by alanine substitution mutagenesis of all basic amino acids in Feldman's region 184-198. A G88 variant truncated beyond residue 198, G58, and G58 fully alanine substituted in the region 184-198, G58A6, bound to HEp-2 cells about half as well and 100-fold less well than G88, respectively. G88 and all alanine substitution mutants of G88 inhibited RSV plaque formation by 50% (ID(50)) at concentrations of approximately 50 nM; the ID(50) of G58 was approximately 425 nM while G58A6 had an ID(50) >1600 nM. These data show that the G HBD includes as much as residues 187-231, that there is redundancy beyond the previously described HBD, and that the cell-binding and virus infectivity-blocking functions of these recombinant G proteins were closely linked and required at least one HBD. PMID- 14551821 TI - Development of a novel quantitative real-time RT-PCR assay for the simultaneous detection of all serotypes of foot-and-mouth disease virus. AB - Foot-and-mouth disease virus (FMDV) spreads extremely fast and the need for rapid and robust diagnostic virus detection systems was obvious during the recent European epidemic. Using a novel real-time RT-PCR system based on primer-probe energy transfer (PriProET) we present here an assay targeting the 3D gene of FMDV. The assay was validated for the efficacy to detect all known FMDV serotypes. The test method was linear over a range of at least 7 orders of magnitude and the detection limit was below the equivalent of 10 genomic copies. Analysing recent African probang samples the method was able to detect FMDV in materials from both cattle and buffalo. When compared to traditional virus cultivation the virus detection sensitivity was similar but the RT-PCR method can provide a laboratory result much faster than virus cultivation. The real-time PCR method confirms the identity of the amplicon by melting point analysis for added specificity and at the same time allows the detection of mutations in the probe region. As such, the described new method is suitable for the robust real-time detection of index cases caused by any serotype of FMDV. PMID- 14551822 TI - Bovine herpesvirus 1 interferes with TAP-dependent peptide transport and intracellular trafficking of MHC class I molecules in human cells. AB - Bovine herpesvirus 1 (BoHV-1), the cause of infectious bovine rhinotracheitis and infectious pustular vulvovaginitis in cattle, establishes a lifelong infection, despite the presence of antiviral immunity in the host. BoHV-1 has been shown to elude the host immune system, but the viral gene products responsible for this interference have not yet been identified. Studies aiming at the identification of BoHV-1-encoded immune evasion genes have been hampered by the lack of bovine specific immunological reagents. Some of the immune evasion molecules identified for other herpesviruses are host species specific; others can act across the species barrier. In this study, experiments were performed to investigate whether BoHV-1 can infect human cells and interfere with antigen processing and presentation in these cells. A human melanoma cell line, Mel JuSo, appeared to be permissive for BoHV-1 infection. BoHV-1 induced expression of major viral glycoproteins at the surface of these cells and produced progeny virus up to 10(5) plaque forming units per ml. BoHV-1 infection resulted in impaired intracellular transport of human MHC class I molecules and inhibition of human TAP. These data indicate that the BoHV-1-encoded molecule(s) that block antigen presentation in bovine cells are able to interact with homologous components of the human MHC class I presentation pathway. The fact that immune evasion by BoHV 1 can be studied in human cells will facilitate the identification of the BoHV-1 gene products involved in this process. Moreover, the data presented here suggest that the BoHV-1 encoded inhibitors of antigen presentation represent potential immune suppressive agents for use in humans. PMID- 14551823 TI - Molecular evidence supporting the classification of Hosta virus X as a distinct species of the genus. AB - Apotexvirus, Hosta virus X (HVX-Kr), causing mosaic and mottle symptoms was isolated from hosta plants ( Hosta spp.) in Korea. The 3'-terminal 2,711 nucleotides excluding the poly (A) tail were determined and shown to include the partial viral replicase, triple gene block (TGB) 1 (26 kDa), TGB2 (13 kDa), TGB3 (8 kDa), and 23 kDa coat protein (CP) and the 3'-nontranslated region (NTR), typical of potexviruses. The CP gene of the type isolate of HVX (HVX-U) was amplified by RT-PCR and its nucleotide sequence was determined. The CPs of HVX-Kr and HVX-U had 100% and 98.9% identical amino acids and nucleotides, respectively. Most of the regions of the genome HVX had over 50% nucleotide identical to other sequenced potexviruses. This is the first report of sequence information of HVX and molecular evidence supporting the virus as a distinct species of the genus Potexvirus. PMID- 14551824 TI - Molecular characterization of squash leaf curl Yunnan virus, a new begomovirus and evidence for recombination. AB - Virus isolate Y23V, obtained from squash showing leaf curl symptoms in Yunnan, China, was readily differentiated from four studied Chinese begomovirus isolates in reactions with a set of monoclonal antibodies raised against begomoviruses. The complete nucleotide sequence (2714 nts) of the DNA-A-like molecule of Y23V was determined. The DNA-A of Y23V is most closely related to that of tomato yellow leaf curl Thailand virus-[1] (TYLCTHV-[1]) (84% sequence identity). However, the AC1 and AC4 gene of Y23V DNA-A resembled to Pepper leaf curl virus from Bangladesh (PepLCBDV). The DNA-A of Y23V has three distinct regions: the region from 74-2071 nts is 95% identical to TYLCTHV-[1] excluding a 27 nt deletion; the following 386 nts are 91% identical to PepLCBDV and the rest of the DNA-A is not closely related to any reported begomovirus. Y23V, therefore, is considered to have arisen by recombination. The 84% sequence identity of Y23V with TYLCTHV-[1] allows Y23V to be considered as a distinct begomovirus species, for which the name squash leaf curl Yunnan virus (SLCYNV) is proposed. PMID- 14551825 TI - Nucleotide sequence and genome organization of Cucurbit yellow stunting disorder virus RNA1. AB - The complete nucleotide sequence of Cucurbit yellow stunting disorder virus (CYSDV) RNA1, a member of the Crinivirus genus in the Closteroviridae, was determined. CYSDV RNA1 is 9126 nucleotides long and contains two overlapping open reading frames (ORFs) that encode the replication module, consisting of the putative papain-like cysteine proteinase, methyl transferase, helicase, and polymerase domains, a small 5 kDa hydrophobic protein and two further downstream ORFs potentially encoding proteins respectively 25 and 22 kDa in size. The genomic position and homology of the four domains comprising the replication module appear to be similar for all sequenced criniviruses but there is divergence in the downstream ORFs, in terms of number, size, position and sequence homology. PMID- 14551826 TI - Viruses in subgroup 2 of the genus Ilarvirus share both serological relationships and characteristics at the molecular level. AB - Sequence data have been determined for 5 members of subgroup 2 of the genus Ilarvirus. These data support the known serological relationships among accepted members of this group and indicate that the ilarvirus Hydrangea mosaic virus (HdMV) is an isolate of Elm mottle virus (EMoV). The close relationships between members of this subgroup, exhibited through the coat proteins coded on RNA 3, extend to the other genomic molecules. Primers designed from the sequences of RNA 1 and RNA 2 of EMoV amplified fragments from all other subgroup 2 viruses but not from other ilarviruses. Although closely related, members of this subgroup occur naturally in distinctly different host species. The possible origins of the viruses are discussed in relation to similarities among the genomic molecules, in particular RNA 3. PMID- 14551828 TI - Negative impacts of a vertebrate predator on insect pollinator visitation and seed output in Chuquiraga oppositifolia, a high Andean shrub. AB - Studies on plant-pollinator interactions have largely neglected the potential negative effects of the predators of pollinators on seed output, even though anti predatory behaviour of pollinators may affect visitation patterns, pollen transfer, and therefore potentially, plant reproductive output. We tested the hypothesis that the presence of lizards and insectivorous birds, by reducing pollinator visitation, can have significant negative effects on seed output in the insect-pollinated, genetically self-incompatible lower alpine Andean shrub, Chuquiraga oppositifolia (Asteraceae). The lower alpine belt supports a high density of territorial Liolaemus (Tropiduridae) lizards and low shrubs interspersed among rocks of varying sizes, the latter inhabited by lizards and commonly used by flycatchers Muscisaxicola (Tyrannidae) as perching sites. In a 2x2 factorial predator-exclusion experiment, visitation rates of the most frequent pollinators of C. oppositifolia (the satyrid butterfly Cosmosatyrus chilensis and the syrphid fly Scaeva melanostoma), the duration of pollinator visits, and seed output, were 2-4 times greater when lizards were excluded, while birds had no effect. In a natural experiment, visits by S. melanostoma were 9 times shorter, and pollinator visitation rates of C. chilensis and S. melanostoma, and C. oppositifolia seed output were 2-3 times lower on shrubs growing adjacent to lizard-occupied rocks compared to those growing distant from rocks. Our results, verified for additional Andean sites, suggest that lizard predators can alter the behaviour of pollinators and elicit strong top-down indirect negative effects on seed output. Such effects may be especially important in high alpine plant communities, where pollinator activity can be low and erratic, and pollen limitation has been reported. PMID- 14551829 TI - [Demands for a new, patient oriented rehabilitation]. AB - A new orientation of rehabilitation in Germany has been initiated by the social political representatives in legislation and government, based on conceptualizations of self help movements in Europe and Germany. The goal is to form a new rehabilitation which is dedicated to participation and partnership of citizens with disability ("Nothing about us without us"). Empowerment is envisaged and resources supplied by legislation to allow to implement these new concepts (personal budget). To guarantee these new rights they have been codified in a new, 9th book of the German social code (Sozialgesetzbuch IX, SGB IX). This new perspective gives rise to major evolutionary impulses in rehabilitation, notably disabled persons' participation in self-assessment, in formulating their rehabilitation goals, determining and controlling the rehabilitation process, terminating rehabilitation (co-signing their discharge reports), as well as evaluating the outcome (quality control of rehabilitation results). They have the right to get all their medical information (right of informational independence). Seven guidelines of people with disability are formulated to support implementation of the new rehabilitation. PMID- 14551827 TI - Cervical myelopathy: clinical and neurophysiological evaluation. AB - The overall frequency of troublesome neck pain is estimated to be about 34%, and it was observed that the frequency of complaints lasting 1 month or longer was higher in women than in men. The prevalence increased with age, with regard to both pain duration and chronic pain. Approximately 14% of a randomly selected population meets the criterion for chronic neck pain: complaints lasting more than 6 months. Epidemiologic data substantiate the importance of morphologic, age related changes of the cervical spine; however, the incidence and prevalence of cervical myelopathy is not known. It could be that the structural transformation of the intervertebral disc, the uncovertebral processes and the zygapophyseal joints is a process accompanied by disturbed function that ultimately not only induces pain, but can lead to narrowing of spinal canal, with symptoms and signs of cervical myelopathy. For a diagnosis of radicular and myelopathic syndromes, the functional and neurological examination is enhanced by neurophysiological assessment. Electromyography (EMG) performed with needle electrodes is the oldest method for diagnosing nerve root compression and anterior horn cell syndromes, and is claimed to have no false-positive results. For cervical myelopathy, as a routine examination sensory evoked potentials (SEPs) by stimulation of tibial nerve and motor evoked potentials (MEPs) from the upper and lower extremities are recommended. PMID- 14551830 TI - [Initial status of patients and effects of rehabilitation after stroke--analysis of a patients' and a physicians' questionnaire in three neurological rehabilitation centres with a follow-up after 6 months]. AB - Rehabilitation after stroke has to face specific problems when treating patients with more or less severe disabilities in cognition and communication. Correspondingly, stroke rehabilitation takes a special position within the larger field of rehabilitation, and relatively little is known outside the neurological scientific community about the status of patients at admission, the case mix in the centres and the short- and medium-term effects of rehabilitation. The present study describes in some detail the initial status in unselected samples of consecutive patients (n = 768) from three neurological rehabilitation centres. The description shows a very inconsistent picture in all centres, ranging from patients with no neurological deficits to patients needing intensive care. Across the centres, we found remarkable differences in case mix. In order to measure the effects of rehabilitation after stroke, an instrument was developed that combines a physicians' questionnaire aiming at an assessment of the severely disabled cases with a patients' questionnaire for the less severe cases for which the physicians' questionnaire would show "ceiling effects" so that improvements could no longer be depicted. The application of the instrument showed that about 50 % of the sample were not capable of answering the patients' questionnaire. For the patients with neurological deficits, the functional parameters of the physicians' questionnaire showed significant improvements at discharge that can be interpreted as "strong" effects (effect sizes 1.0-1.3). For the patients with less severe deficits (and usually in later stages of the rehabilitation process), the patients' questionnaire showed "strong" improvements on the somatic and psychosocial scales both at discharge and 6 months later. On the functional scales, however, only small improvements were found. Finally, predictors could be identified that explain a large amount of the variance for length of stay (R(2) =.42) as well as for the effects of rehabilitation (R(2) =.74). When comparing effects across rehabilitation units with differences of case mix, these predictors should be statistically controlled in order to assure fair comparisons. PMID- 14551831 TI - [Relevance of medical rehabilitation in disease management programmes]. AB - Disease management programmes will increasingly be introduced in Germany due to the new risk adjustment scheme. The first disease management programmes started in 2003 for breast cancer and diabetes mellitus type II. German rehabilitation will have to face several challenges. Disease management programmes are strongly based on the notion of Evidence so that proof of the efficacy of a care giving task should be present. Verification of the evidence of the specifically German rehabilitation treatments must therefore be given. However, integration of rehabilitation in disease management programmes could lead to changes in the alignment of German rehabilitation. The essence of German rehabilitation, notably its holistic approach, could get lost with integration in disease management programmes. PMID- 14551832 TI - [Disease profiles and therapy effects of patients in mother-child rehabilitation centres]. AB - Within the setting of external quality management of mother-child rehabilitation centres the short-term and medium-term therapy effects as well as the disease profiles of patients were analysed. This paper is focused on the changing of the main symptoms among the mothers. The patients' state of health is essentially characterized by physical and psychological exhaustion as well as by multimorbidity. It is strongly associated with the mothers' current situation of living. Before intervention the patients' health status was significantly impaired. Directly after the treatment high or very high therapy effects were found. All indicators are still higher after 6 months than they were at the beginning of the treatment. The results of a quasi-control population give evidence that these effects are caused by the treatment. First trends about the children treated show that behaviour problems can be reduced and the quality of life enhanced. PMID- 14551833 TI - [Potentials of the Berufliche Trainingszentren, BTZ--participant centered vocational rehabilitation of persons with chronic mental illness]. AB - Potentials, opportunities of performance--a spectrum of alternatives which can be activated when required as a single measure, varied or in total, low- or high powered, specifically adjusted or compressed. There are dynamism and power to it! This sounds and feels different from what is demanded all over the world: "standards" or even "minimum standards", minimum quality standards and minimum performance levels, standard equipment, reducing things to the lowest possible common denominator, uniformity, not meeting clients' needs but serving all in the same way, not in the sense of optimum but of minimum. These short polemics are to elucidate the fundamental difference between client focus and client orientation. At certain times in the process of vocational rehabilitation, men and women with mental problems or illness need individually conceived, if necessary improvised, individually composed and well-coordinated services. What kind of men and women we cope with is stated in the chapter on "Target groups", neither completely nor exhaustively--there are no limits to the fantasy and the opportunities of development of clients. In a second major chapter, an attempt is made to show- regarding to some carefully, not arbitrarily selected but not all--the criteria for whom the Berufliche Trainingszentren (BTZ) have developed adequate, client focused, individual potentials. PMID- 14551834 TI - [Concepts in early planning for vocational rehabilitation in schizophrenia patients during acute care]. AB - The particular relevance of employment for financial independence, acquisition of social skills and, hence, an increase in self-confidence is undeniable, and has for several years now been increasingly considered an essential component of care notably in psychiatric disorders. Vice versa, unemployment is deemed a potential risk factor with regard to poorer therapeutic response and social integration as well as a higher incidence of crises in persons with mental illness. Schizophrenia is among the psychiatric disorders very often resulting in unemployment and subsequent early pensioning. Planning for vocational rehabilitation early-on during acute care therefore is considered of greatest importance especially in this patient group. This article gives a brief overview of the main concepts drawn on in preparing the vocational rehabilitation of patients with Schizophrenia, focussing in particular on their relevance for daily practice and the possibilities for evaluating their effectiveness. PMID- 14551835 TI - [On the German Orthopedics Congress 2003. Geriatric orthopaedics]. PMID- 14551837 TI - [Femoral periprosthetic bone remodelling to the proximal femur after implantation of custom made anatomic and standard straight stem hip prostheses]. AB - AIM: As a result of stress shielding bone resorption occurs around straight femoral stems following total hip replacement (THR). The question arises whether this pattern of periprosthetic bone loss is altered with use of custom made anatomic femoral stems. METHOD: DEXA method was used to examine proximal femora of two groups of patients after cementless THR. Data of 16 patients with a standard straight femoral stem and 15 patients with a custom made anatomic stem were acquired at 1 week and 2, 4, 6, 9, 12 and 24 months postoperatively. Periprosthetic bone density was recorded in regions of interest (ROI). RESULTS: Similar pattern of periprosthetic bone changes were seen in femora with straight and anatomic hip stems at 24 months postoperatively. Femoral bone loss, up to 36 % in the calcar ROI, was seen with straight and anatomic stems. CONCLUSION: Custom designed anatomic femoral hip stems were unable to prevent periprosthetic bone resorption. However it is concluded, that implantation of custom made stems in grossly distorted femoral anatomy induces transmission of forces similar to standard femoral stems implanted in normal medullary anatomy. PMID- 14551836 TI - [Optimizing primary total hip replacement--a technique to effect saving of manpower]. AB - PURPOSE: Development of a standardized surgical technique for total hip replacement thereby saving manpower (one assistant) by using a retractor system. METHOD: Total hip replacement is performed with the patient in a true lateral position on a tunnel cushion. By means of a direct lateral approach the pelvitrochanteric muscles are partially detached using an omega-shaped cut. The Bookwalter retractor is fixed dorsally on the operating table. The ring is centered keeping the greater trochanter in the middle. The Hohmann retractors are fixed to the ring to sufficiently expose the acetabulum. To insert the femoral stem the ring needs to be opened dorsally and the patient's leg is bent 90 degrees in the hip and the knee over the tunnel cushion. The muscles inserting at the greater trochanter are retracted by a separate Hohmann retractor with weight. In a case control study with matched pairs the patients treated with this technique were compared with those treated in supine position with the transgluteal approach. The number of assistants required and the operating time were assessed. RESULTS: All the hip replacements with the patient in side position were performed with one assistant, in supine position with two assistants. The operating time did not differ significantly (supine position 110 min/side position 112 min). The complication rate in both groups was comparable (one secondary wound healing, one transient ischalgia). CONCLUSION: The process of total hip replacement can be optimized. The described technique allows to spare one surgical assistant without prolonging the operating time. PMID- 14551838 TI - [Bone adaptation changes mechanical stress in the femur--a prospective two years follow up after Huft-TEP implantation]. AB - AIM: Periprosthetic adaptive bone remodeling after total hip arthroplasty (THA) has been frequently simulated in computer models, combining bone remodeling theory with finite element analysis. Unfortunately, there is still a lack of clinical validation data. The collection of prospective volumetric bone density data with a clinical computerized tomography study after THA was previously conducted. The objective of the study presented here is an additional evaluation of femoral strain. METHOD: In a prospective clinical trial with 7 female patients after THA computerized tomography (CT) and finite element (FE) modeling was patient specifically combined immediately after surgery, as well as at three and 24 months postoperatively. Mechanical strain was expressed by strain energy density. RESULTS: Corresponding to a bone density decrease, a decrease of the femoral strain energy density was observed during the two year follow-up after surgery (approximately 20 %). The predominant change occurred within the first three months and was found to be distally stronger than proximally. CONCLUSION: As far as we are aware, this is the first combination of fully prospective 3D CT density data in vivo with patient-specific finite element modeling. The assessment of mechanical strain data during a follow-up trial should be a new approach for analyzing hip stems in clinical biomechanics. A future confirmative study based on a statistical case number calculation would increase the evidence of the results presented here. PMID- 14551839 TI - [Intraoperative 3D reconstruction of the PMMA plug for computer-assisted revision of total hip arthroplasty based on 2D X-ray images]. AB - AIM: Computer-assisted cement removal out of the femoral medullary canal requires the exact definition of the plug's shape within a 3D coordination system. Aiming at a sufficiently precise reconstruction based on segmentation of just a few 2D X ray images a special mathematic procedure is needed for automatic surface interpolation. The geometric specification of the resulting virtual model should take into account the characteristic geometry of the navigated tools in order to achieve the best possible removal. METHOD: Studies were performed on anatomic specimens of entire human femora, that underwent cemented THA before being cut every 5 mm. The cross-sections were scanned at the high resolution of 600 dpi. Segmentation of the cement was performed with the help of a virtual deformable template and was both used for simulation of X-ray projections from various points of view and for validation of the reconstructed 3D model. By this means systematic errors such as those possible during X-ray acquisition, tracking or segmentation could be avoided and the precision of the procedure could be measured exclusively. RESULTS: With increasing number of X-rays the distance from the reconstructed 3D model to the original could continuously be reduced. Using only two x-rays a maximum error was measured with 6.5 mm, whereas 5 pictures taken from different angles showed to be enough to ensure an error below 1 mm in the distal part of the femur. By the use of 6 or more pictures no significant improvement could be attained. CONCLUSION: The innovative procedure is essential for future 2D image-based fluoroscopic navigation of PMMA removal and bears the options of computer-controlled and robotic material working, respectively. PMID- 14551840 TI - [[F-18]-fluorodeoxyglucose (FDG) positron emission tomography (PET) as a diagnostic for hip endoprosthesis loosening]. AB - AIM: The purpose of this study was to examine the FDG-PET in respect of its diagnostic valency with regard to septic/aseptic loosening of lower limb prostheses. METHOD: 28 patients with 41 lower limb prostheses were examined by means of FDG-PET to evaluate septic/aseptic loosening of their hip prostheses. Therefore, a differentiated FDG-PET result interpretation subdivided into five categories was developed. The final diagnosis was based on operative findings with following culture and histological outcome. RESULTS: The worked-out categories showed a high agreement to the intraoperative macroscopic and histological results (n = 23 correctly positive, n = 1 false positive, n = 1 correctly negative and n = 3 false negative). CONCLUSION: With a subtly differentiated interpretation (categories I - V) of the qualitative glucose metabolism safe statements can be made regarding septic/aseptic endoprostheses loosening. This was impressively confirmed by the agreement of the FDG results with the histological results. PMID- 14551841 TI - [Stage diagnostics for postinfection revision of hip and knee replacement: value of laboratory parameters and antigranulocyte scintigraphy]. AB - AIM: In the case of two-stage infect revision arthroplasties of hip and knee joint, of primary resection arthroplasties and before implantation of arthroplasties after septic arthritis the accuracy of preoperative laboratory parameters and antigranulocyte scintigraphy was analysed. Furthermore, we stained the intraoperatively taken joint synovial samples with hematoxylin-eosin and also with antibodies against human neutrophil elastase in order to investigate if immunohistological examination provides further or different information. METHOD: In 24 patients with intraoperative verification of infection we calculated sensitivity, specificity, positive and negative predictive values for laboratory tests, antigranulocyte scintigraphy and the most suitable combination of both. The joint synovial samples stained with antibodies against human neutrophil elastase were compared to those stained with hematoxylin-eosin according to the criteria of Mirra. RESULTS: We observed the best results for the combination of C reactive protein and antigranulocyte scintigraphy with a sensitivity of 1, a specificity of 0.92, a positive predictive value of 0.75 and a negative predictive value of 1. No additional or different information was observed by the immunohistological stained samples. CONCLUSION: Stage diagnostic using C-reactive protein and antigranulocyte scintigraphy provides accurate information to assess the status of infection before hip and knee replacement after infect revision. Additional immunohistological staining besides the routinely taken hematoxylin eosin staining of joint synovial samples is not recommended. PMID- 14551842 TI - [Femoral fissure after cement restrictor implantation]. AB - Cement restrictors are accepted parts of modern cementing technique and reduce the risk of revision. The restrictor should seal the intramedullary cavity to ensure sustained pressurization and optimal cement penetration into cancellous bone. Significant complications after use of a cement restrictor are rarely described. Dislocation and cement leakage are well known problems. We describe the occurrence of a femoral fissure after insertion of a resorbable Gelatine cement restrictor. PMID- 14551843 TI - [Patella resurfacing in patients without substantial retropatellar knee pain symptoms?]. AB - OBJECTIVE: The aim is of this prospective randomised study was to investigate the necessity of resurfacing the patella in combination with total knee arthroplasty in patients without or mild anterior knee pain before undergoing total knee replacement. METHODS: Between May 1999 and May 2000 fifty patients were enrolled in a prospective, randomised study. All patients received the same posterior cruciate-sparing total knee replacement and were randomised to treatment with and without resurfacing of the patella. Inclusion criteria were primary osteoarthritis of the knee, preoperatively no pain when the patella was shifted during clinical examination, a maximal grade III radiological degeneration of the patella according to Sperner et al. and at most a mild anterior knee pain in preoperative interview. Evaluations consisted of the determination of the Knee Society clinical score, the completion of a patient satisfaction questionnaire, and radiographic assessment basing on the Knee Society roentgenographic evaluation and scoring system. All patients were examined preoperatively and 3, 6, and 12 months postoperatively. RESULTS: In all postoperative examinations patients with patella resurfacing demonstrated a higher overall Knee Society score. At month 6 the difference was statistically significant. The patient satisfaction questionnaire demonstrated no significant difference between both groups. However, regarding to relief of anterior knee pain and improvement of pain in general patients with nonresurfacing were less satisfied at all follow ups. Furthermore, after 12 months all answers to the patient satisfaction questionnaire of the resurfaced group were better. Patella-associated revisions were performed in two patients without resurfacing. Radiographic analysis demonstrated no loosening, fracture, subluxation or dislocation. CONCLUSION: The presented study demonstrated after one year follow-up a better functional result, a higher degree of contentment and fewer complications after total knee replacement with patella resurfacing in patients without or mild preoperative anterior knee pain. PMID- 14551844 TI - [Arthroscopic menisectomy in older patients: assessing health-related quality of life]. AB - AIM: Little information is available on partial arthroscopic menisectomy in patients 60 years of age or older. Due to the co-morbidity of meniscal tears and osteoarthritis in this age group, the effectiveness of this procedure is controversial. The purpose of this study was to evaluate the outcome of this procedure in terms of function and mobility in patients 60 years of age or older. METHOD: Between 1990 and 1999, 1920 arthroscopies of the knee were performed in a single primary care institution, and 51 patients, 60 years of age or older, had a partial arthroscopic menisectomy done. Outcomes were rated using the SF-36 Health Survey and a specially designed questionnaire including items of published scores. RESULTS: The mean age at surgery was 67 years (range: 60.3 - 78.9) and the mean follow-up was 5 years (range: 2 - 12). Due to persistent pain, 4 patients underwent a second operation including knee replacement in 3 cases and a high tibial osteotomy in 1 case. 41 patients (87.1 %) stated the arthroscopy had improved their quality of life. In terms of pain and daily life activities, the majority of the subjects showed an improvement after surgery. The SF-36 showed no clinically relevant difference compared to the control in all but one subscale. CONCLUSION: Despite of the presence of osteoarthritis in this age group, arthroscopic partial menisectomy can improve pain, function and mobility in the mid-term course. PMID- 14551845 TI - [Massive arthrofibrosis of the knee joint after carbon fibre rod implantation. Two cases]. AB - Summary. In the treatment of cartilage damage the use of carbon rods is getting more and more popular. The papers are promising. There are no reports about significant complications. The following cases show a post operative enormous increasing discomfort. PMID- 14551846 TI - [Comparison of dorso-lateral and dorso-ventral stabilization procedures in the treatment of vertebral fractures]. AB - AIM: The radiological and clinical outcome of surgical treated thoracolumbal fractures were followed up after 6.7 years. METHOD: The study encompassed 97 patients. The degrees of the bony deformation and the local kyphosis were measured on the lateral view X-ray at 4 different time points: post trauma, after the operation, before implant removal and at the follow up examination. The clinical outcome was evaluated by the Oswestry score. RESULTS: 74 fractures were treated with a fixateur intern and a dorsal fusion by apposition of autologous bone postero-laterally. 23 fractures were fused in a combined dorso-ventral manner by intervertebral fusion with tricortical autologous bone. The lateral X rays showed a loss of correction up to 48 % in the dorsal fused group and 10 % loss of correction in the combined group. The local kyphosis increased up to 84 % in the dorsal operated group due to degeneration of the injured disk. The clinical Oswestry score showed no significant difference in both groups. CONCLUSION: The additional ventral surgery should be evaluated carefully. PMID- 14551847 TI - [Coccygectomy in the treatment of coccygodynia]. AB - AIM: Excision of the coccyx for the treatment of therapy-resistant coccygodynia is a disputable management option. Due to the low morbidity only few studies concerning the long-term follow-up after coccygectomy exist. The aim of this study is a retrospective analysis of our patients surgically managed for coccygodynia and a critical review of the results obtained in comparison to the literature. METHOD: 12 patients with complete radiographic and clinical data were included in the study. The average age of patients at the time of surgery was 43.3 years (11 - 75 years). The average follow-up was 9.8 years (2 - 16 years). As suggested by Hambly (1989) the clinical result was assessed according to postoperative pain status and subjective patient satisfaction. RESULTS: 9 of 12 patients regarded the surgical intervention as a success and claimed that they would repeat the procedure (75 %). Three patients did not show marked improvement after coccygectomy. All patients (n = 6) surgically managed for traumatically induced coccygodynia had a positive result, while only 3/6 patients treated for idiopathic coccygodynia reported that symptoms were postoperatively reduced. CONCLUSION: According to our results and review of those documented in the literature, excision of the coccyx for the treatment of coccygodynia, after all conservative treatment options have been exhausted, seems a justifiable alternative. Patients with a history suggestive of traumatically induced coccygodynia are more likely to benefit from coccygectomy. PMID- 14551848 TI - [Calculation of hallux bending moments from plantar pressure distribution measurement]. AB - PURPOSE: The following study aims to demonstrate the bending stress on a hallux valgus during normal gait. The bending axis was related to the position of an open wedge osteotomy. The moments were calculated from plantar pressure measurements. METHOD: In the gait analysis laboratory plantar pressure distribution measurement was done with healthy adult volunteers during barefoot gait. From the plantar pressure distribution the vertical forces were derived just for the region hallux metatarsal I. The following calculation of the bending stress regarding to the hypothetic region of the osteotomy was done by integrating the pressure distribution multiplicated with the distance to the osteotomy. The measurements are compared with theoretical values derived from the anatomy of the foot under some plausible assumptions. RESULTS: The results of the mean values of the bending stress were 15.8 +/- 4.8 Nm for the female subjects and 21.6 +/- 5.6 Nm for the male subjects. The amount of the bending moments depended with low significance on the weight. More important is the individual type of gait. CONCLUSION: The measurements show the size of bending stress the plate stabilizing the osteosynthesis has to resist under unfavorable circumstances (for example if the plantar foot muscles are not adequately activated) and if no external stabilization (for example, cast) is used. PMID- 14551849 TI - [The Weil osteotomy in metatarsalgia]. AB - AIM: The purpose of the prospective study is to evaluate the results of Weil osteotomy in painful overloaded central forefoot with dislocation of the MTP joint. METHOD: 19 left and 16 right feet underwent the Weil osteotomy. In 10 patients the procedure was performed on both feet. In 15 cases only one metatarsal was corrected (the 2 (nd) in 12 and the 3 (rd) in 3 cases), in 17 cases both the 2 (nd) and the 3 (rd) metatarsals and in 4 cases also with the 4 (th) one together. The simultaneous correction of hallux valgus was performed in 19 forefeet. The mean follow-up was 31 months (range, 19 to 41 months). RESULTS: All patients were satisfied with the result of the operation. The results were excellent in 29 cases (83 %) and good in 6 cases (17 %). The plantar callus disappeared in 27 feet (77 %) and decreased in 8 feet (23 %). In one patient the new callus formed under the head of the neighbouring metatarsal bone. The sufficient MTP joint reduction was found in 31 cases (89 %). CONCLUSION: It is possible with the Weil osteotomy to correct the metatarsal length accurately according to the preoperative planning. This results in the prevention of new callus formation under the neighbouring metatarsal head. Stable internal fixation allows the early mobilisation of the MTP joint. PMID- 14551850 TI - [Long-term sequels after BCG osteomyelitis]. AB - AIM: The aim of the retrospective study was to record and evaluate the long-term sequels in patients after their bone growth end, who went through the BCG osteomyelitis in their childhood. PATIENTS AND METHODS: Five patients were chosen for the study, in them the BCG osteomyelitis was proven according to Foucard's criteria, and who had undergone a Moscow-type BCG vaccination in the maternity hospital. They have been diagnosed, treated and followed-up till the end of their bone growth, in the course of 18 years. RESULTS: On evaluation of the long-term sequels, the shortening of limbs, deviation of knee joint axis, muscular hypotrophy of the limbs, and residual changes on the X-ray after an epiphyseal infection focus were found. CONCLUSION: Post-vaccination specific bone infection has been treated in a complex way. The long-term sequels that were found after the bone growth end, have not been considered for correction surgery any longer. PMID- 14551851 TI - [The concept of liberty in Jaspers' psychopathology]. AB - The debate on conceptional problems represents a fundamental and inevitable challenge also for contemporary biological psychiatry. Especially questions concerning liberty, loss of liberty and regaining liberty are relevant for daily psychiatric practice. This study attempts to critically and systematically investigate the answers given in the philosophy and psychopathology of Karl Jaspers. Thereby, the key term "Grenzsituation" (border situation) plays a significant role. The interpretation of psychiatric disorder as an exceptional state of existence, possibly converting "Alltagssituationen" (situations of daily life) to "Grenzsituationen", sheds new light on Jaspers' thoughts about the concept of liberty which, thus, turn out to be of crucial relevance for the necessary discussions of ethical principles in the era of molecular psychiatry. PMID- 14551852 TI - [Expressed emotion and the outcome of child and adolescent psychiatric disorders]. AB - OBJECTIVE: The review discusses empirical research on the association of "Expressed Emotion" (EE) and the outcome of child and adolescent psychiatric disorders. METHOD: A comprehensive search in the databases Medline, PsycINFO and PSYNDEX was performed. Moreover, publications cited in the reference sections were taken into consideration. RESULTS: Empirical data suggest that EE is a predictor of the outcome of depressive and eating disorders in childhood and adolescence, but not of behavioural disorders and ADHD. There are no studies on EE and its influence on anxiety disorders and - surprisingly - only two case studies on EE and schizophrenia in childhood and adolescence. Across psychiatric groups there is an association between behavioural disturbances of the child and parental criticism and an association of child anxiety and parental emotional over involvement. Data suggest that EE is not a stable variable. There is some evidence that a high-EE-status of parents transforms from a state into a trait variable. CONCLUSION: Further studies on the predictive validity of EE for the outcome of child adolescent psychiatric disorders are warranted, as well as studies on variables of the parents and their offspring, which are associated with EE and with the entrenchment of EE with the progress of the disorder. Child and adolescent psychiatrists may contribute to a large research area with studies on the early development of high-EE. The mutual goal is the development of effective early family interventions. PMID- 14551853 TI - [Factor structure and validity of a german version of the barratt impulsiveness scale]. AB - OBJECTIVE: Impulsive traits are key characteristics in a number of psychiatric disorders and are part of the normal behavior spectrum. The BIS-5 is an instrument developed to assess impulsivity. The aim of this study is to evaluate the BIS-5 in two German psychiatric inpatient samples and healthy controls proving the originally proposed four-factor structure as well as convergent and discriminate validity. METHODS: 159 alcohol-dependent subjects and 77 suicidal inpatients were recruited in an University psychiatric hospital. 182 healthy subjects were recruited from town community. BIS-5 items were translated and back translated. Principal component analysis with oblique rotation was conducted in the whole group. Furthermore, the discriminate and convergent validity of the BIS 5 was evaluated by correlation with other instruments measuring impulsive traits and comparing sample subgroups. RESULTS: A two-factor solution could be identified in this German sample. Alcohol-dependent individuals showed significantly higher factor 1 values compared to suicidal patients. The group of suicidal patients had higher scores in factor 2 compared to controls. Factor 1 correlated most significantly with extraversion-related personality traits while factor 2 showed significant relationships with irritability and neuroticism. CONCLUSIONS: A two-factor solution may be more appropriate in using the BIS-5 scale in German samples. These two factors might reflect different aspects of impulsive behavior and might be useful to characterize impulsive behavior in psychiatric and non-psychiatric samples. PMID- 14551854 TI - [Hyperlipidemia - side-effect of "Atypical" neuroleptics?]. AB - After the introduction of the so-called 'atypical antipsychotics' some reports concerning hyperlipidemia observed in patients treated with these drugs have been published. The studies and case reports available up to now were reviewed. The available data show that hyperlipidemia particularly occurred in patients receiving clozapine, olanzapine and also quetiapine. Predominately elevated serum levels of triglycerides have been reported. The underlying pathomechanism still remains widely unclear. Since hyperlipidemia is an important symptom of the so called 'metabolic syndrome' which is often associated with severe complications like cardial and vascular diseases, more attention should be paid to hyperlipidemia as a potential side effect of antipsychotics. PMID- 14551855 TI - [Awareness of deficits in patients with dementia of the Alzheimer type]. PMID- 14551856 TI - [Diagnostics and therapy of myositis]. PMID- 14551857 TI - DDW reports 2003 Orlando: reflux disease and Barrett's esophagus. PMID- 14551858 TI - DDW reports 2003 Orlando: capsule endoscopy. PMID- 14551859 TI - Treatment of bleeding colonic diverticula by endoscopic band ligation: an in-vivo and ex-vivo pilot study. AB - BACKGROUND AND STUDY AIMS: Angiographic and surgical therapy are standards of care for persistent diverticular bleeding. Colonoscopic intervention using epinephrine injection, multipolar electrocautery, and placement of an Endoclip has not gained widespread acceptance due to concerns about complications,and the widespread management of severe lower gastrointestinal bleeding by surgeons and interventional radiologists. The utility of colonoscopic band ligation for control of diverticular bleeding was evaluated both in vivo and ex vivo. PATIENTS AND METHODS: Endoscopic band ligation of diverticula was performed on surgical resection specimens and in patients with actively bleeding colonic diverticula. RESULTS: In the in-vivo study, active diverticular bleeding was completely controlled in four patients by endoscopic band ligation. In two cases, a visible vessel was seen on the everted and banded diverticulum. Procedure time ranged from 45 to 140 min. The total lengths of hospital stays for the four patients were 2, 6, 14, and 35 days. The long hospital stays (> 7 days)were associated with non-gastrointestinal co-morbidity. There were no acute complications of band ligation. No rebleeding or need for surgery occurred during a follow-up period of 12 months in any of the patients. In the ex-vivo study, 11 diverticula were successfully everted and banded in five of nine surgical specimens (one right colon and four left colons). Mucosa was identified in all of the "banded" segments. Ten of 11 ligated diverticula revealed evidence of blood vessels or submucosal tissue. The presence of subserosal fat was suggested in three of the 11 "banded" segments, and none of the ex-vivo ligated diverticula contained muscularis propria or serosal involvement. There was no evidence of perforation. CONCLUSIONS: Both in-vivo and ex-vivo data suggest that endoscopic band ligation may be a safe and effective therapy for actively bleeding colonic diverticula. PMID- 14551860 TI - Risk factors for pancreatitis following endoscopic retrograde cholangiopancreatography: a meta-analysis. AB - BACKGROUND AND STUDY AIM: The identification of factors which increase the risk of acute pancreatitis, the most common and most severe complication of endoscopic retrograde cholangiopancreatography (ERCP), is of considerable importance. However, in four large prospective studies, the reported incidence of identified risk factors for post-ERCP pancreatitis was found to be variable. The aim of this study was to identify the most important risk factors for post-ERCP pancreatitis. MATERIAL AND METHODS: We performed a meta-analysis of 15 prospective clinical studies, selected out of 52 reviewed articles, in which risk factors for pancreatitis were identified. Of the 14 risk factors identified, five were patient characteristics and nine were related to the endoscopic technique. RESULTS: When patient-related risk factors were analysed, the relative risk for suspected sphincter of Oddi dysfunction was 4.09 (95 % CI 3.37 to 4.96; P < 0.001); for female gender, 2.23 (95 % CI 1.75 to 2.84, P < 0.001); and for previous pancreatitis, 2.46 (95 % CI 1.93 to 3.12, P < 0.001). Two endoscopy related factors were confirmed: for precut sphincterotomy the relative risk was 2.71 (95 % CI 2.02 to 3.63, P < 0.001); for pancreatic injection the relative risk was 2.2 (95 % CI 1.6 to 3.01, P < 0.001). CONCLUSIONS: This meta-analysis provides a clear basis for planning pharmacological studies or studies of new endoscopic techniques in patients at high risk of developing post-ERCP pancreatitis. Knowing which patients are most at risk may also make it easier to decide who should be considered unsuitable for same-day discharge. PMID- 14551861 TI - Standardization and quality of endoscopy text reports in ulcerative colitis. AB - BACKGROUND AND STUDY AIMS: The text report is the primary tool for documenting endoscopic findings but there is no consensus on the content and structure of these reports. Therefore, at four Norwegian hospitals, the content of endoscopy reports concerning ulcerative colitis was assessed. Quality indices for the medical history of active ulcerative colitis and endoscopic signs of inflammation were determined, as well as technical items in the report. The effect of structured compared with free-text reporting was evaluated. MATERIALS AND METHODS: Endoscopy reports in 445 cases of ulcerative colitis were retrieved. Two of the hospitals used a semi-structured computerized documentation system, and two hospitals used transcription-based free-text reports. RESULTS: A substantial amount of information was missing in the majority of the reports. Individual endoscopic signs of inflammation were defined in 27 % - 77 % of the reports. Various clinical symptoms of active ulcerative colitis were defined in 1 % - 44 % of the reports. We observed a reminder effect of structured systems in that they prompted more informative reports. There was a tendency towards better free-text documentation in the transcription-based systems than in the free text of the semi-structured ones. CONCLUSIONS: There is a potential for improving the content, completeness and standardization of endoscopy reports. Standardization efforts may be a part of the solution. PMID- 14551862 TI - A modified method of botulinum toxin injection in patients with achalasia: a pilot trial. AB - BACKGROUND AND AIMS: Injection of botulinum toxin (BT) from direct vision into the lower esophageal sphincter (LES) lowers its basal tone and improves symptomatology in most of the patients with achalasia. We hypothesized that the effect could be improved by better degree of LES infiltration by toxin administered from both prograde and retrograde views. The aim of the study was to investigate the feasibility, safety and efficacy of this modified method of intrasphincteric BT injection in patients with achalasia. PATIENTS AND METHOD: Sixteen patients with achalasia were treated with BT injection. Hundred units of BT (Botox) were diluted with 4 ml of normal saline. Aliquots of 0.5 ml were injected into four quadrants of the LES from retrovision and then into each quadrant from direct vision. The patients were followed up for a median of 25.5 months (range 19-31). RESULTS: No serious adverse events were noted. All patients responded well to the injection within one week and 3 patients (18.7 %) experienced an early relapse. The remaining 13 patients were classified as responders. After a single BT injection, 11 responders reported a relapse and 2 patients remained asymptomatic. The median symptom-free interval was 17 months (8 28). Five patients with a relapse underwent BT reinjection. Three of them remained asymptomatic and two experienced the second relapse. After BT reinjection, the median symptom-free interval was 16 months (10-19). All other patients with a relapse and without BT reinjection were treated with either balloon dilatation or surgery and are currently asymptomatic. CONCLUSION: Combining injection of BT into the LES from both direct vision and retrovision was feasible, safe and produced a rapid response which was sustained for more than 1 year in the majority of patients. This method of BT administration might be superior to the traditional injection from direct view only, and a randomized and prospective study comparing those techniques of administration should be performed. PMID- 14551863 TI - Long-term follow-up of large flat colorectal tumors resected endoscopically. AB - BACKGROUND AND STUDY AIM: When a large flat colorectal tumor is excised by piecemeal endoscopic resection, it is not possible to obtain suitable specimens for histopathological examination to assess whether the resection has been complete. We prospectively analysed follow-up colonoscopy examinations of endoscopic polypectomy resection sites for residual lesions. PATIENTS AND METHODS: A total of 24 patients with large flat colorectal tumors were treated using an endoscopic submucosal saline injection technique. The resection site was prospectively examined for residual lesions using a magnifying colonoscope, at 3, 6, 12 and 24 months postoperatively. RESULTS: None of the 5 patients who underwent en bloc resection exhibited residual lesions postoperatively. Of the 19 patients who underwent piecemeal resection, one died of an asthma attack and 18 were followed up. Residual lesions were detected in four of these 18 patients (three adenomas and one cancer), which were resected endoscopically. When the tumor was resected endoscopically en bloc and was judged histologically to be completely resected, residual lesions were not detected at follow-up. However, residual lesions were detected in 22.2 % of patients 3 months after initial resection when polyps were resected piecemeal. After treatment, residual lesions were detected in two patients (11.1 %) at 24 months. CONCLUSION: After piecemeal endoscopic resection for large flat colorectal tumors, it is necessary to follow up and remove residual lesions endoscopically every 3 months until they all are removed. PMID- 14551864 TI - Newly developed Barrett's esophagus after subtotal esophagectomy. AB - BACKGROUND AND STUDY AIMS: More detailed information regarding the early mucosal events that lead to intestinal metaplasia would be very beneficial for understanding the pathogenesis of Barrett's esophagus (BE). Gastroesophageal reflux and duodenogastroesophageal reflux play a major role in the pathogenesis of Barrett's esophagus. The aim of this study was to investigate the prevalence of newly developed BE in patients who had previously undergone a subtotal esophagectomy - a clinical condition characterized by the absence of a lower esophageal sphincter and massive gastroesophageal reflux. PATIENTS AND METHODS: A retrospective examination was carried out on all patients who underwent subtotal esophagectomy (n = 87) listed in our institution's computer files from 1995 to 2000. Twenty-one patients were excluded due to missing data or no upper gastrointestinal endoscopy after surgery. RESULTS: Based on the Savary-Miller classification, 47 patients developed either type I (n = 2), II (n = 8), III (n = 11) or IV (n = 26) esophagitis after surgery. Newly developed BE was observed in nine patients (13.5 %) after subtotal esophagectomy (median time to diagnosis: 489 days, range 43 - 1172). None of the patients had persistent BE immediately after surgery, and two of the patients with newly developed BE had had no history of BE before surgery or at the time of surgery. Proton-pump inhibitor therapy after surgery and neoadjuvant chemotherapy did not appear to influence the development of BE after subtotal esophagectomy. CONCLUSIONS: Newly developed BE after subtotal esophagectomy may provide further insights into the early mucosal events that lead to intestinal metaplasia and into the roles of gastroesophageal and duodenoesophageal reflux in the pathogenesis of BE. PMID- 14551865 TI - Ampullary carcinoid tumor presenting as acute pancreatitis in a patient with von Recklinghausen's disease: case report and review of the literature. AB - A patient with von Recklinghausen syndrome presented with multiple attacks of pancreatitis. Evaluation using computed tomography, endoscopic retrograde cholangiopancreatography, and endosonography revealed an ampullary carcinoid. The patient underwent a pylorus-sparing Whipple resection, and is doing well after 30 months of follow-up. This is the second reported case of ampullary carcinoid presenting with pancreatitis in a patient with von Recklinghausen syndrome. PMID- 14551866 TI - Y-nitinol airway stent for management of central airway compression due to metastatic colon cancer. AB - Tumor masses in the area between the esophagus and the tracheobronchial tree can lead to complications involving both systems, mainly strictures and compressions. Malignant esophageal strictures are nowadays often treated by insertion of a metal stent which, however, can cause airway compression especially in the proximal area. We present here a new method of creating a Y-stent out of two self expandable tracheal nitinol stents, utilizing fiber bronchoscopy, in a 55-year old woman with advanced colon cancer metastastic to the mediastinum. The endo-Y stent technique can be performed with the patient under sedation and having topical anesthesia. The opening through which the second tracheal stent must be placed for the Y construction is created by laser. In this case, the patient suffered from airway compression which was efficiently relieved by this method. Within a short time the endo-Y-stent provides effective restoration and maintenance of airway patency in patients with tumor compression in the region of the esophagus and airway, and in those with airway compression following esophageal stenting. Expertise in both stent implantation and laser application is, however, mandatory. PMID- 14551867 TI - Locally advanced small-bowel adenocarcinoma missed primarily by capsule endoscopy but diagnosed by push enteroscopy. AB - Wireless-capsule-endoscopy is a new painless method that is able both to visualize the entire small bowel and to detect even small lesions. We report here the case of a patient in whom a locally advanced small-bowel adenocarcinoma was initially missed on capsule endoscopy, but was diagnosed by subsequent push enteroscopy. Capsule endoscopy was carried out in a 47-year-old patient with a history of obscure gastrointestinal bleeding, iron-deficiency anemia, and a lack of symptoms suggestive of stenosis. The capsule imaging revealed angiodysplasias in the jejunum, but no other abnormalities. Push enteroscopy was carried out to allow argon plasma coagulation treatment of the angiodysplasias that had been detected; it revealed a polypoid tumor 20 mm in diameter at 150 cm from the incisors, with the capsule endoscope still located proximal to the tumor and with its optical dome turned towards the push enteroscope. Clinical staging and subsequent surgical resection showed a locally advanced adenocarcinoma (pT4, pN0 (0/7), pMx, G3).[nl]Small-bowel tumors within the reach of push enteroscopy may be missed by capsule endoscopy. Although wireless capsule endoscopy is a major advance in the investigation of the small bowel, well-designed clinical studies still need to define the precise algorithm for diagnostic work-up of suspected small-bowel diseases. PMID- 14551868 TI - Small-bowel tumors diagnosed by wireless capsule endoscopy: report of five cases. AB - Tumors of the small bowel, both benign and malignant, are relatively uncommon. As the symptoms are vague and conventional diagnostic tests are unsatisfactory, these tumors often present a clinical, radiological, and endoscopic challenge. We report here on five patients in whom small-bowel tumors were diagnosed using wireless capsule endoscopy. The indications for capsule endoscopy were: obscure gastrointestinal bleeding in four patients (one jejunal capillary hemangioma, one ileal hemangiosarcoma, and two jejunal gastrointestinal stromal tumors) and chronic abdominal pain in one patient (ileal carcinoid). Wireless capsule endoscopy, a new endoscopic method, promises to improve the diagnosis of deep small-bowel pathology. PMID- 14551869 TI - ESGE/ESGENA technical note on cleaning and disinfection. PMID- 14551870 TI - Cleaning and disinfection in Europe according to the Endoscopic Societies' Guidelines. PMID- 14551871 TI - Gastritis cystica polyposa found in an unoperated stomach: an unusual case treated by endoscopic polypectomy. PMID- 14551872 TI - Endoscopic management of complications related to esophageal stenting by repositioning the stent without extraction or re-stenting. PMID- 14551873 TI - Distal migration of a self-expandable metal oesophageal stent, presenting as constipation. PMID- 14551874 TI - Second-look endoscopy during percutaneous endoscopic gastrostomy by passing a laryngofiberscope through the inserted feeding tube. PMID- 14551875 TI - A rare cause of sigmoid obstruction in a 77-year-old patient. PMID- 14551876 TI - Genotypic differences in the Chlamydia pneumoniae tyrP locus related to vascular tropism and pathogenicity. AB - Chlamydia pneumoniae is an obligate intracellular pathogen that causes respiratory infections and has been associated with cardiovascular disease. We compared respiratory and cardiovascular isolates to find genetic differences associated with pathogenicity. A polymorphic region encoding a tyrosine/tryptophan permease was found to differ between disease isolates. Respiratory strains contained multiple copies of the tyrP gene, and vascular strains contained a single copy. Single-nucleotide polymorphism analysis revealed the duplication to be a phylogenetically old event. Gene amplification was associated with higher mRNA levels and higher uptake of the substrate tyrosine, indicating an amino-acid transport-related phenotype associated with the tyrP genotype. Vascular strains, despite their reduced ability to transport tyrosine, do not appear to have a reduced growth rate in vitro. We hypothesize that the important difference between strains of vascular and respiratory origin may lie in the increased tendency of vascular strains to elicit persistent infection that is triggered by amino-acid starvation. PMID- 14551877 TI - Nuclear factor-kappaB activation in endothelium by Chlamydia pneumoniae without active infection. AB - Causative molecular mechanisms accounting for the potential link between Chlamydia pneumoniae and atherosclerosis are unknown. Formalin and heat inactivated C. pneumoniae activated the transcription factor nuclear factor (NF) kappaB in cultured porcine endothelium and up-regulated the expression of E selectin messenger RNA and protein. This up-regulation was abolished by an IkappaB super-repressor, an NF-kappaB-specific inhibitor. Live bacteria are not necessary for the activation of endothelial NF-kappaB, and C. pneumoniae may contribute to atherogenesis without active infection. PMID- 14551878 TI - Selective up-regulation of matrix metalloproteinase-9 expression in human erythema migrans skin lesions of acute lyme disease. AB - Despite the absence of enzymes that digest extracellular matrix, Borrelia burgdorferi spreads in the skin to form erythema migrans (EM) lesions and then disseminates to other organs. We studied the induction by bacteria of host matrix metalloproteinases (MMPs) in EM skin lesions of patients with acute Lyme disease. In blister fluid from the EM lesions, the expression of MMP-9 was selectively increased by 1900%+/-1037%, compared with blister fluid from the surrounding normal-appearing skin. The expression of all other MMP messenger RNAs was similar in the EM lesions and normal-appearing skin. Selective up-regulation of MMP-9 in the EM lesions was found. Fibroblasts and, to a lesser degree, mononuclear cells were the sources of local MMP-9 production. These results demonstrate specific up regulation of MMP-9 in the EM skin lesions of patients with acute Lyme disease. Bacterial induction of host proteases may play a role in the dissemination of B. burgdorferi. PMID- 14551879 TI - Pathophysiology and prognosis in vietnamese adults with tuberculous meningitis. AB - The pathogenesis of tuberculous meningitis remains unclear, and there are few data describing the kinetics of the immune response during the course of its treatment. We measured concentrations of pro- and anti-inflammatory cytokines in serial blood and cerebrospinal fluid (CSF) samples from 21 adults who were being treated for tuberculous meningitis. CSF concentrations of soluble tumor necrosis factor-alpha receptors and of matrix metalloprotein-9 and its tissue inhibitor were also measured, and blood-brain barrier permeability was assessed by the albumin and IgG partition indices. CSF concentrations of lactate, interleukin-8, and interferon-gamma were high before treatment and then decreased rapidly with antituberculosis chemotherapy. However, significant immune activation and blood brain barrier dysfunction were still apparent after 60 days of treatment. Death was associated with high initial CSF concentrations of lactate, low numbers of white blood cells, in particular neutrophils, and low CSF glucose levels. PMID- 14551880 TI - No evidence for association between the polymorphism in the 3' untranslated region of interleukin-12B and human susceptibility to tuberculosis. AB - Interleukin (IL)-12 plays a pivotal role in cell-mediated immunity to Mycobacterium tuberculosis infection. We tested the association between a biallelic single-nucleotide polymorphism in the 3' untranslated region (UTR) of IL-12B and human susceptibility to tuberculosis (TB), in a population-based case control study of adult patients with TB from 2 ethnicities, African American and white, and in a family-based transmission/disequilibrium study of 60 informative families with at least 1 pediatric patient with TB and 1 heterozygous parent. Our results suggest that IL-12B 3' UTR has no effect or has a negligible effect on human susceptibility to TB. PMID- 14551881 TI - Streptococcus pneumoniae-associated human macrophage apoptosis after bacterial internalization via complement and Fcgamma receptors correlates with intracellular bacterial load. AB - Opsonization enhances Streptococcus pneumoniae-induced human monocyte-derived macrophage (MDM) apoptosis. Both depletion of complement and immunoglobulin from opsonizing serum and blockade of the macrophages CR1, CR3, FcgammaRII, and FcgammaRIII partially decreased MDM apoptosis after S. pneumoniae phagocytosis, and these effects correlated with reduced numbers of internalized bacteria. Chloramphenicol inhibition of protein synthesis by opsonized S. pneumoniae down regulated subsequent MDM apoptosis. Phagocytosis of an unencapsulated mutant of S. pneumoniae resulted in increased MDM apoptosis, in association with enhanced internalization. Caspase inhibition was associated with decreased killing of bacteria. Enhanced induction of apoptosis by opsonized S. pneumoniae is the result of increased intracellular burden of bacteria, rather than of a specific pattern of engagement of complement receptor or FcgammaR. A dynamic interaction between live intracellular bacteria and the host cell is necessary for induction of apoptosis in MDMs, and induction of apoptosis contributes to the host defense against S. pneumoniae. PMID- 14551882 TI - Molecular characterization of serotype III group B-streptococcus isolates causing neonatal meningitis. AB - We studied a collection of 110 serotype III group B streptococcus (GBS) isolates causing neonatal meningitis, by means of both pulsed-field gel electrophoresis (PFGE) with SmaI and Southern hybridization with probes for genes potentially associated with virulence (neuA, cpsA, scpB, and hylB and, for mobile genetic elements [MGEs], GBSi1 and IS1548), in comparison with 44 serotype III GBS isolates colonizing healthy neonates. Using polymerase chain reaction, we assessed both the insertion of MGEs downstream of the scpB gene and the insertion of IS1548 within the hylB gene. PFGE clustered the isolates into 3 main groups. One PFGE group accounted for 80% of typeable cerebrospinal fluid (CSF) isolates, versus 24% of colonization isolates (P=1.8 x 10-9). GBSi1 was found in 67% of CSF isolates and in only 23% of colonization isolates (P=5.3 x 10-7). A 15-kbp SmaI restriction-DNA fragment bearing the neuA gene was significantly associated with CSF isolates (P=1.1 x 10-11). PMID- 14551883 TI - Anthrax toxin induces hemolysis: an indirect effect through polymorphonuclear cells. AB - Anthrax toxin can induce hemolysis in the presence of polymorphonuclear cells (PMNs), an activity primarily mediated by protective antigen, with synergic effects provided by lethal factor and edema factor. Lethal factor and edema factor, individually or in combination, are incapable of lysing red blood cells. The requirement for the presence of PMNs indicates that hemolysis associated with Bacillus anthracis infection is indirect rather than direct, as observed in many other bacterial infections. PMID- 14551884 TI - Effects of total body irradiation and cyclosporin a on the lethality of toxic shock syndrome toxin-1 in a rabbit model of toxic shock syndrome. AB - Toxic shock syndrome (TSS) may be mediated by superantigen-activated T cells, a theory we tested in rabbits, which are more susceptible to the lethal effects of superantigens, such as TSS toxin-1 (TSST-1), than are mice. Rabbits exposed to 10 cGy of total body irradiation exhibited T cell deficiency, with profound depletion of splenic lymphocytes and circulating CD4(+) lymphocytes, as well as an inability to manifest delayed-type hypersensitivity. Nevertheless, these rabbits remained completely susceptible to TSST-1, indicating that TSS can occur in the setting of marked immunosuppression. PMID- 14551885 TI - Virological and immunological impact of tuberculosis on human immunodeficiency virus type 1 disease. AB - Unlike other opportunistic infections associated with human immunodeficiency virus (HIV) type 1, tuberculosis (TB) occurs throughout the course of HIV-1 infection, and, as a chronic infection, its impact on viral activity is sustained. In dually infected subjects, HIV-1 load and heterogeneity are increased both locally and systemically during active TB. Studies over the past decade have indicated that Mycobacterium tuberculosis (MTB) infection supports HIV-1 replication and dissemination through the dysregulation of host cytokines, chemokines, and their receptors. Furthermore, concentrations of HIV-1 inhibitory chemokines are limited during TB and at sites of MTB infection. Cumulatively, these data indicate that TB provides a milieu of continuous cellular activation and irregularities in cytokine and chemokine circuits that are permissive of viral replication and expansion in situ. I address new research that has identified the basis for the augmentation of HIV-1 replication during TB and discuss potential immunotherapies to contain viral expansion during TB. PMID- 14551886 TI - Stable incidence rates of tuberculosis (TB) among human immunodeficiency virus (HIV)-negative South African gold miners during a decade of epidemic HIV associated TB. AB - During the last decade, annual tuberculosis (TB) case-notification rates increased 4-fold, to >4000 cases/100000 person-years, in the study workforce, among whom prevalence of human immunodeficiency virus (HIV) was 30% in 2000. Three separate cohort studies, totalling 6454 HIV-negative participants, were combined and analyzed for time trends. Observed incidence of TB varied between 962 (1991-1994) and 1589 (1999-2000) cases/100000 person-years (P=.17, test for trend). There was, however, a progressive increase in age, and, for each period, older age was associated with increased incidence rates of TB (P<.001). Having adjusted for age differences, there was no significant association between incidence of TB and calendar period (P=.81, test for trend). Relative to 1991 1994, multivariate-adjusted incidence-rate ratios were 0.94, for 1995-1997, 0.96, for 1998-1999, and 1.05, for 1999-2000. Preventing a secondary epidemic of TB among HIV-negative individuals may be achievable with conventional means, even in settings with a high burden of HIV-associated TB. PMID- 14551887 TI - Prevalence and correlates of untreated human immunodeficiency virus type 1 infection among persons who have died in the era of modern antiretroviral therapy. AB - We evaluated all human immunodeficiency virus (HIV)-related deaths over the period 1 January 1995-31 December 2001 in a Canadian province in which all HIV care and antiretroviral therapy are provided free of charge. Persons who had received antiretroviral drugs before death were compared with those who had died without ever receiving HIV treatment, by fitting a logistic model. Overall, 1239 deaths were attributed to HIV infection during the study period. Of these, 406 (32.8%) occurred among persons who had never received any HIV treatment. In adjusted analyses, aboriginal ethnicity, female sex, and lower median income were negatively associated with receiving HIV treatment before death. Furthermore, among the 833 individuals who received treatment before death, only 379 (45.5%) received antiretroviral medication >or=75% of the time during their first year receiving therapy. The data demonstrate the need for novel interventions to expand HIV care to specific populations. PMID- 14551889 TI - Modeling a safer smallpox vaccination regimen, for human immunodeficiency virus type 1-infected patients, in immunocompromised macaques. AB - We have modeled smallpox vaccination with Dryvax (Wyeth) in rhesus macaques that had depletion of CD4(+) T cells induced by infection with simian immunodeficiency virus or simian/human immunodeficiency virus. Smallpox vaccination induced significantly larger skin lesions in immunocompromised macaques than in healthy macaques. Unexpectedly, "progressive vaccinia" was infrequent. Vaccination of immunocompromised macaques with the genetically-engineered, replication-deficient poxvirus NYVAC, before or after retrovirus infection, was safe and lessened the severity of Dryvax-induced skin lesions. Neutralizing antibodies to vaccinia were induced by NYVAC, even in macaques with severe CD4(+) T cell depletion, and their titers inversely correlated with the time to complete resolution of the skin lesions. Together, these results provide the proof of concept, in macaque models that mirror human immunodeficiency virus type 1 infection, that a prime-boost approach with a highly attenuated poxvirus followed by Dryvax increases the safety of smallpox vaccination, and they highlight the importance of neutralizing antibodies in protection against virulent poxvirus. PMID- 14551888 TI - Sequence conservation and antibody cross-recognition of clade B human immunodeficiency virus (HIV) type 1 Tat protein in HIV-1-infected Italians, Ugandans, and South Africans. AB - We determined immune cross-recognition and the degree of Tat conservation in patients infected by local human immunodeficiency virus (HIV) type 1 strains. The data indicated a similar prevalence of total and epitope-specific anti-Tat IgG in 578 serum samples from HIV-infected Italian (n=302), Ugandan (n=139), and South African (n=137) subjects, using the same B clade Tat protein that is being used in vaccine trials. In particular, anti-Tat antibodies were detected in 13.2%, 10.8%, and 13.9% of HIV-1-infected individuals from Italy, Uganda, and South Africa, respectively. Sequence analysis results indicated a high similarity of Tat from the different circulating viruses with BH-10 Tat, particularly in the 1 58 amino acid region, which contains most of the immunogenic epitopes. These data indicate an effective cross-recognition of a B-clade laboratory strain-derived Tat protein vaccine by individuals infected with different local viruses, owing to the high similarity of Tat epitopes. PMID- 14551890 TI - Hepatitis C and human immunodeficiency virus envelope proteins cooperatively induce hepatocytic apoptosis via an innocent bystander mechanism. AB - We hypothesized that hepatocytes exposed to hepatitis C virus (HCV) and human immunodeficiency virus (HIV) might be injured via an "innocent bystander" mechanism due to cell-surface binding of viral proteins. To assess this, we studied the effects of HCV envelope protein E2 and T-tropic HIV envelope glycoprotein gp120 on hepatocytes and saw potent apoptosis. Either viral protein alone did not induce this effect. HCV E2 and M-tropic HIV gp120 also induced significant apoptosis. Blocking the CXCR4 receptor led to a reduction in apoptosis. HCV E2 and HIV gp120 acted collaboratively to trigger a specific set of downstream signaling events, including up-regulation of the Fas ligand and dephosphorylation of the anti-apoptotic molecule AKT. These results suggest that hepatic injury may occur in HCV/HIV coinfection through the induction of novel downstream signaling pathways and provide a rationale for therapeutic interventions that interfere with specific receptors and signaling molecules. PMID- 14551891 TI - Are polymorphic markers within the alpha-1-antitrypsin gene associated with risk of human immunodeficiency virus disease? AB - We sought to determine whether variants of the human alpha-1-antitrypsin (AAT) gene, also known as "PI," or "SERPINA1," are associated with human immunodeficiency virus (HIV) infection in 2 African-based populations from HIV pandemic sub-Saharan Africa. Eleven commonly occurring African-associated polymorphic markers in the coding and intronic regions of the AAT gene were analyzed via denaturing gradient gel electrophoresis. A significant association between HIV-1 infection and the presence of an allelic variant was observed in the case of the M2 and A332A haplotypes, thus presenting AAT as a potentially novel HIV-1 susceptibility locus. PMID- 14551892 TI - Human immunodeficiency virus type 1 RNA in breast-milk components. AB - We conducted the present study to determine which of the 4 components of breast milk (whole milk, skim milk, lipid layer, and breast-milk cells) had the highest sensitivity and concentration of human immunodeficiency virus (HIV) type 1 RNA burden and to determine biological correlates to these factors. The probability of detection of HIV (sensitivity) and the concentration of HIV-1 RNA were both associated with the choice of milk component, CD4(+) cell count, concentration of blood serum HIV-1 RNA, and the presence of breast inflammation. Whole milk demonstrated higher sensitivity and mean concentration than any other single component. Sensitivity was enhanced by analyzing all 4 components of breast milk. PMID- 14551893 TI - Chimeric live, attenuated vaccine against Japanese encephalitis (ChimeriVax-JE): phase 2 clinical trials for safety and immunogenicity, effect of vaccine dose and schedule, and memory response to challenge with inactivated Japanese encephalitis antigen. AB - ChimeriVax-JE is a live, attenuated vaccine against Japanese encephalitis, using yellow fever (YF) 17D vaccine as a vector. In a double-blind phase 2 trial, 99 adults received vaccine, placebo, or YF 17D vaccine (YF-VAX). ChimeriVax-JE was well tolerated, with no differences in adverse events between treatment groups. Viremias resulting from administration of ChimeriVax-JE and YF-VAX were of short duration and low titer; 82 (94%) of 87 subjects administered graded doses (1.8 5.8 log(10)) of ChimeriVax-JE developed neutralizing antibodies. A second dose, administered 30 days later, had no booster effect. Previous inoculation with YF did not interfere with ChimeriVax-JE, but there was a suggestion (not statistically significant) that ChimeriVax-JE interfered with YF-VAX administered 30 days later. A separate study explored immunological memory both in subjects who had received ChimeriVax-JE 9 months before and in ChimeriVax-JE-naive subjects challenged with inactivated mouse-brain vaccine (JE-VAX). Anamnestic responses were observed in preimmune individuals. ChimeriVax-JE appears to be a safe vaccine that provides protective levels of neutralizing antibody after a single dose. PMID- 14551894 TI - Prevention of increasing rates of treatment failure by combining sulfadoxine pyrimethamine with artesunate or amodiaquine for the sequential treatment of malaria. AB - Combination antimalarial therapy may delay the spread of drug resistance, but clinical data supporting this notion are limited. For 1 year, we studied Ugandan children who were treated for uncomplicated malaria with sulfadoxine pyrimethamine (SP), SP + amodiaquine (AQ), or SP + artesunate (AS). We compared treatment responses and the prevalence of resistance-conferring mutations of new infections with those of recrudescent infections due to parasites that survived prior treatment. Recrudescent infections were associated with the selection of SP resistance-conferring mutations in all treatment groups, but responses to repeat therapy differed. Compared with initial treatments, treatment of recrudescent infections was associated with a higher rate of treatment failure (hazard ratio [HR], 2.44; P=.01), for the SP group, but with a lower rate of treatment failure (HR, 0.40; P=.08), for the SP + AS group. Treatment failure in the SP + AQ group was uncommon, limiting the analysis of recrudescent parasites. Our results suggest that the use of combination antimalarial therapy in Africa may slow the spread of drug-resistant malaria and prolong the therapeutic life span of available treatment regimens. PMID- 14551895 TI - Safety and immunogenicity of DNA/modified vaccinia virus ankara malaria vaccination in African adults. AB - The present study is an investigation of the safety and immunogenicity of DNA and modified vaccinia virus Ankara (MVA) candidate vaccines, each encoding the malaria DNA sequence multiple epitope-thrombospondin related adhesion protein (ME TRAP), against Plasmodium falciparum. DNA ME-TRAP and MVA ME-TRAP are safe and immunogenic for effector and memory T cell induction. MVA ME-TRAP, with or without prior DNA ME-TRAP immunization, was more immunogenic and more cross reactive in malaria-exposed individuals than in malaria-naive individuals, a finding suggesting that recombinant MVA vaccines are particularly promising for the development of a malaria vaccine for exposed populations. Both CD4(+) and CD8(+) T cells were induced by these vaccines. PMID- 14551896 TI - Pathogen genotyping in polyclonal infections: application of a fluorogenic polymerase-chain-reaction assay in malaria. AB - Pathogen genotyping of polyclonal infections is limited by 2 major drawbacks: (1). how to establish whether multiple mutations detected in 1 gene belong to the same clone and (2). how to evaluate the proportion of different genotypes in a given sample. For drug-resistance genotyping in Plasmodium falciparum malaria, we address these problems by using a fluorogenic assay that combines fluorescence resonance energy transfer, between fluorophores present on a probe and a polymerase-chain-reaction primer, and a melt-curve analysis. We demonstrate that this tool allows a more accurate insight into the P. falciparum populations present in complex biological samples. PMID- 14551897 TI - Protection against cutaneous leishmaniasis in outbred vervet monkeys using a recombinant histone H1 antigen. AB - Infection with Leishmania major parasites results in the development of cutaneous ulcerative lesions on the skin. We investigated the protective potential of a single, recombinant histone H1 antigen against cutaneous leishmaniasis in an outbred population of vervet monkeys, using Montanide adjuvant. Protection was assessed by challenging the animals with a mixture of vector sand fly salivary gland lysate and a low dose of in vitro-derived parasites, thus more closely mimicking natural infection induced by L. major. The course of infection in immunized monkeys was compared with that of animals that had healed from a primary infection and were immune. The monkeys immunized with recombinant histone H1 showed a reduced development of lesion size, compared with controls. Our study therefore illustrates the potential use of histone H1 as a vaccine candidate against cutaneous leishmaniasis in humans. PMID- 14551898 TI - Changing transmission fitness of drug-resistant human immunodeficiency virus against a background of evolving antiretroviral therapy. PMID- 14551900 TI - Burden of malaria during pregnancy in areas of stable and unstable transmission in Ethiopia during a nonepidemic year. PMID- 14551902 TI - Accounting for linkage in family-based tests of association with missing parental genotypes. AB - In studies of complex diseases, a common paradigm is to conduct association analysis at markers in regions identified by linkage analysis, to attempt to narrow the region of interest. Family-based tests for association based on parental transmissions to affected offspring are often used in fine-mapping studies. However, for diseases with late onset, parental genotypes are often missing. Without parental genotypes, family-based tests either compare allele frequencies in affected individuals with those in their unaffected siblings or use siblings to infer missing parental genotypes. An example of the latter approach is the score test implemented in the computer program TRANSMIT. The inference of missing parental genotypes in TRANSMIT assumes that transmissions from parents to affected siblings are independent, which is appropriate when there is no linkage. However, using computer simulations, we show that, when the marker and disease locus are linked and the data set consists of families with multiple affected siblings, this assumption leads to a bias in the score statistic under the null hypothesis of no association between the marker and disease alleles. This bias leads to an inflated type I error rate for the score test in regions of linkage. We present a novel test for association in the presence of linkage (APL) that correctly infers missing parental genotypes in regions of linkage by estimating identity-by-descent parameters, to adjust for correlation between parental transmissions to affected siblings. In simulated data, we demonstrate the validity of the APL test under the null hypothesis of no association and show that the test can be more powerful than the pedigree disequilibrium test and family-based association test. As an example, we compare the performance of the tests in a candidate-gene study in families with Parkinson disease. PMID- 14551906 TI - Genetically modified corn--environmental benefits and risks. PMID- 14551903 TI - A functional analysis of the spacer of V(D)J recombination signal sequences. AB - During lymphocyte development, V(D)J recombination assembles antigen receptor genes from component V, D, and J gene segments. These gene segments are flanked by a recombination signal sequence (RSS), which serves as the binding site for the recombination machinery. The murine Jbeta2.6 gene segment is a recombinationally inactive pseudogene, but examination of its RSS reveals no obvious reason for its failure to recombine. Mutagenesis of the Jbeta2.6 RSS demonstrates that the sequences of the heptamer, nonamer, and spacer are all important. Strikingly, changes solely in the spacer sequence can result in dramatic differences in the level of recombination. The subsequent analysis of a library of more than 4,000 spacer variants revealed that spacer residues of particular functional importance are correlated with their degree of conservation. Biochemical assays indicate distinct cooperation between the spacer and heptamer/nonamer along each step of the reaction pathway. The results suggest that the spacer serves not only to ensure the appropriate distance between the heptamer and nonamer but also regulates RSS activity by providing additional RAG:RSS interaction surfaces. We conclude that while RSSs are defined by a "digital" requirement for absolutely conserved nucleotides, the quality of RSS function is determined in an "analog" manner by numerous complex interactions between the RAG proteins and the less-well conserved nucleotides in the heptamer, the nonamer, and, importantly, the spacer. Those modulatory effects are accurately predicted by a new computational algorithm for "RSS information content." The interplay between such binary and multiplicative modes of interactions provides a general model for analyzing protein-DNA interactions in various biological systems. PMID- 14551907 TI - Out of the way: how the next copyright revolution can help the next scientific revolution. PMID- 14551908 TI - The roles of APC and Axin derived from experimental and theoretical analysis of the Wnt pathway. AB - Wnt signaling plays an important role in both oncogenesis and development. Activation of the Wnt pathway results in stabilization of the transcriptional coactivator beta-catenin. Recent studies have demonstrated that axin, which coordinates beta-catenin degradation, is itself degraded. Although the key molecules required for transducing a Wnt signal have been identified, a quantitative understanding of this pathway has been lacking. We have developed a mathematical model for the canonical Wnt pathway that describes the interactions among the core components: Wnt, Frizzled, Dishevelled, GSK3beta, APC, axin, beta catenin, and TCF. Using a system of differential equations, the model incorporates the kinetics of protein-protein interactions, protein synthesis/degradation, and phosphorylation/dephosphorylation. We initially defined a reference state of kinetic, thermodynamic, and flux data from experiments using Xenopus extracts. Predictions based on the analysis of the reference state were used iteratively to develop a more refined model from which we analyzed the effects of prolonged and transient Wnt stimulation on beta catenin and axin turnover. We predict several unusual features of the Wnt pathway, some of which we tested experimentally. An insight from our model, which we confirmed experimentally, is that the two scaffold proteins axin and APC promote the formation of degradation complexes in very different ways. We can also explain the importance of axin degradation in amplifying and sharpening the Wnt signal, and we show that the dependence of axin degradation on APC is an essential part of an unappreciated regulatory loop that prevents the accumulation of beta-catenin at decreased APC concentrations. By applying control analysis to our mathematical model, we demonstrate the modular design, sensitivity, and robustness of the Wnt pathway and derive an explicit expression for tumor suppression and oncogenicity. PMID- 14551910 TI - Genome-wide RNAi of C. elegans using the hypersensitive rrf-3 strain reveals novel gene functions. AB - RNA-mediated interference (RNAi) is a method to inhibit gene function by introduction of double-stranded RNA (dsRNA). Recently, an RNAi library was constructed that consists of bacterial clones expressing dsRNA, corresponding to nearly 90% of the 19,427 predicted genes of C. elegans. Feeding of this RNAi library to the standard wild-type laboratory strain Bristol N2 detected phenotypes for approximately 10% of the corresponding genes. To increase the number of genes for which a loss-of-function phenotype can be detected, we undertook a genome-wide RNAi screen using the rrf-3 mutant strain, which we found to be hypersensitive to RNAi. Feeding of the RNAi library to rrf-3 mutants resulted in additional loss-of-function phenotypes for 393 genes, increasing the number of genes with a phenotype by 23%. These additional phenotypes are distributed over different phenotypic classes. We also studied interexperimental variability in RNAi results and found persistent levels of false negatives. In addition, we used the RNAi phenotypes obtained with the genome-wide screens to systematically clone seven existing genetic mutants with visible phenotypes. The genome-wide RNAi screen using rrf-3 significantly increased the functional data on the C. elegans genome. The resulting dataset will be valuable in conjunction with other functional genomics approaches, as well as in other model organisms. PMID- 14551911 TI - Heterochromatin dynamics. PMID- 14551912 TI - Microarray analysis: genome-scale hypothesis scanning. PMID- 14551913 TI - V(D)J recombination and the evolution of the adaptive immune system. PMID- 14551914 TI - Neuroscience networks: data-sharing in an information age. PMID- 14551915 TI - Digital evolution. PMID- 14551917 TI - Developmental origin and evolution of bacteriocytes in the aphid-Buchnera symbiosis. AB - Symbiotic relationships between bacteria and insect hosts are common. Although the bacterial endosymbionts have been subjected to intense investigation, little is known of the host cells in which they reside, the bacteriocytes. We have studied the development and evolution of aphid bacteriocytes, the host cells that contain the endosymbiotic bacteria Buchnera aphidicola. We show that bacteriocytes of Acyrthosiphon pisum express several gene products (or their paralogues): Distal-less, Ultrabithorax/Abdominal-A, and Engrailed. Using these markers, we find that a subpopulation of the bacteriocytes is specified prior to the transmission of maternal bacteria to the embryo. In addition, we discovered that a second population of cells is recruited to the bacteriocyte fate later in development. We experimentally demonstrate that bacteriocyte induction and proliferation occur independently of B. aphidicola. Major features of bacteriocyte development, including the two-step recruitment of bacteriocytes, have been conserved in aphids for 80-150 million years. Furthermore, we have investigated two cases of evolutionary loss of bacterial symbionts: in one case, where novel extracellular, eukaryotic symbionts replaced the bacteria, the bacteriocyte is maintained; in another case, where symbionts are absent, the bacteriocytes are initiated but not maintained. The bacteriocyte represents an evolutionarily novel cell fate, which is developmentally determined independently of the bacteria. Three of five transcription factors we examined show novel expression patterns in bacteriocytes, suggesting that bacteriocytes may have evolved to express many additional transcription factors. The evolutionary transition to a symbiosis in which bacteria and an aphid cell form a functional unit, similar to the origin of plastids, has apparently involved extensive molecular adaptations on the part of the host cell. PMID- 14551925 TI - PLoS biology--we're open. PMID- 14551921 TI - What controls variation in human skin color? PMID- 14551926 TI - Why PLoS became a publisher. PMID- 14551927 TI - [Selection and identification of human keratinocyte stem cells in vitro]. AB - OBJECTIVE: To investigate the selection and identification of human keratinocyte stem cells(KSC) in vitro. METHODS: According to the characteristics of KSC which can adhere to extracellular matrix very fast, we selected 3 groups of different time(5 minutes, 20 minutes and 60 minutes) and unselected as control group. And the cells were identified by monoclone antibody of beta 1-integrin and cytokeratin 19 (Ck19), then the image analysis was done. Furthermore we analyzed the cultured cells with flow cytometer(FCM) and observed the ultrastructure of the cell by transmission electron microscope(TEM). RESULTS: The cell clones formed in all groups after 10 to 14 days, while the cells of 5 minute group grew more slowly than those of the other groups, however, the clones of this group were bigger. The expression of beta 1-integrin and Ck19 were found in all groups. The positive rate of beta 1-integrin was significant difference between 5 minute group and the other groups (P < 0.05). And the expression of Ck19 was no significant difference between 5 minute group and 20 minute group(P > 0.05), and between 60 minute group and control group. But significant difference was observed between the former and the later groups(P < 0.05). The result of FCM showed that most cells of the 5 minute group lied in G1 period of cell cycle, which was different from those of the other groups. At the same time, the cells of 5 minute group were smaller and contained fewer organelles than those of the other groups. CONCLUSION: The above results demonstrate that the cells of 5 minute group have a slow cell cycle, characteristics of immaturity, and behaving like clonogenic cells in vitro. The cells have the general anticipated properties for KSC. So the KSC can be selected by rapid attachment to extracellular matrix and identified by monoclone antibody of beta 1-integrin and Ck19. PMID- 14551916 TI - Candidate gene association study in type 2 diabetes indicates a role for genes involved in beta-cell function as well as insulin action. AB - Type 2 diabetes is an increasingly common, serious metabolic disorder with a substantial inherited component. It is characterised by defects in both insulin secretion and action. Progress in identification of specific genetic variants predisposing to the disease has been limited. To complement ongoing positional cloning efforts, we have undertaken a large-scale candidate gene association study. We examined 152 SNPs in 71 candidate genes for association with diabetes status and related phenotypes in 2,134 Caucasians in a case-control study and an independent quantitative trait (QT) cohort in the United Kingdom. Polymorphisms in five of 15 genes (33%) encoding molecules known to primarily influence pancreatic beta-cell function-ABCC8 (sulphonylurea receptor), KCNJ11 (KIR6.2), SLC2A2 (GLUT2), HNF4A (HNF4alpha), and INS (insulin)-significantly altered disease risk, and in three genes, the risk allele, haplotype, or both had a biologically consistent effect on a relevant physiological trait in the QT study. We examined 35 genes predicted to have their major influence on insulin action, and three (9%)-INSR, PIK3R1, and SOS1-showed significant associations with diabetes. These results confirm the genetic complexity of Type 2 diabetes and provide evidence that common variants in genes influencing pancreatic beta-cell function may make a significant contribution to the inherited component of this disease. This study additionally demonstrates that the systematic examination of panels of biological candidate genes in large, well-characterised populations can be an effective complement to positional cloning approaches. The absence of large single-gene effects and the detection of multiple small effects accentuate the need for the study of larger populations in order to reliably identify the size of effect we now expect for complex diseases. PMID- 14551928 TI - [Experimental study on effect of platelet-rich plasma in repair of bone defect]. AB - OBJECTIVE: To study the effect of platelet-rich plasma in the repair of bone defect. METHODS: Segmental bone defects of 1 cm were created in the mid-upper part of bilateral radius of 24 New Zealand white rabbits. One side was randomly chosen as the experimental side, which was filled with artificial bone with platelet-rich plasma (PRP). The other side filled with artificial bone without PRP as the control. After 2, 4, 8 and 12 weeks of implantation, the gross, radiological, histological observations, and computer graphic analysis were performed to investigate the bone healing of the defect in both sides. RESULTS: Two weeks after operation, new bone and fibrous tissue formation in both the experimental and the control sides were observed only in the areas adjacent to the cut ends of the host bone, but the amount of new tissue in the experimental side was much more than that in the control side. In the 4th and 8th weeks, the surface of the artificial bone was covered with a large amount of new bones, the artificial bone was bridged tightly with the host bone by callus in the experimental side, while new bone was limited mainly in the cut ends and was less mature in the control side. In the 12th weeks, bone defects were entirely healed in the experimental side, which were covered completely with cortical bone, while new bone formation was only observed in the ends of artificial bone and there were not continuous bone callus on the surface in the control side. CONCLUSION: Artificial bone with PRP is effective in the repair of segmental bone defects, and PRP could improve the healing of bone defect. PMID- 14551929 TI - [The role of Smads and related transcription factors in the signal transduction of bone morphogenetic protein inducing bone formation]. AB - OBJECTIVE: To clarify the mechanisms of the signal transduction of bone morphogenetic proteins (BMPs) inducing bone formation and to provide theoretical basis for basic and applying research of BMPs. METHOD: We looked up the literature of the role of Smads and related transcription factors in the signal transduction of BMPs inducing bone formation. RESULTS: The signal transduction processes of BMPs included: 1. BMPs combined with type II and type I receptors; 2. the type I receptor phosphorylated Smads; and 3. Smads entered the cell nucleus, interacted with transcription factors and influenced the transcription of related proteins. Smads could be divided into receptor-regulated Smads (R Smads: Smad1, Smad2, Smad3, Smad5, Smad8 and Smad9), common-mediator Smad (co Smad: Smad4), and inhibitory Smads (I-Smads: Smad6 and Smad7). Smad1, Smad5, Smad8, and probable Smad9 were involved in the signal transduction of BMPs. Multiple kinases, such as focal adhesion kinase (FAK), Ras-extracellular signal regulated kinase (ERK), phosphatidylinositol 3-kinase (PI3K), and Akt serine/threonine kinase were related to Smads signal transduction. Smad1 and Smad5 related with transcription factors included core binding factor A1 (CBFA1), smad-interacting protein 1 (SIP1), ornithine decarboxylase antizyme (OAZ), activating protein-1 (AP-1), xenopus ventralizing homeobox protein-2 (Xvent-2), sandostatin (Ski), antiproliferative proteins (Tob), and homeodomain-containing transcriptian factor-8 (Hoxc-8), et al. CBFA1 could interact with Smad1, Smad2, Smad3, and Smad5, so it was involved in TGF-beta and BMP-2 signal transduction, and played an important role in the bone formation. Cleidocranial dysplasia (CCD) was thought to be caused by heterozygous mutations in CBFA1. The CBFA1 knockout mice showed no osteogenesis and had maturational disturbance of chondrocytes. CONCLUSION: Smads and related transcription factors, especially Smad1, Smad5, Smad8 and CBFA1, play an important role in the signal transduction of BMPs inducing bone formation. PMID- 14551930 TI - [Repair of bone defect with compound of coralline hydroxyapatite porous, fibrin sealant and Staphylococcus aureus injection]. AB - OBJECTIVE: To investigate the ability of repairing bone defect with the compound of coralline hydroxyapatite porous (CHAP), fibrin sealant(FS) and staphylococcus aureus injection (SAI), and the feasibility to use the compounds as bone substitute material. METHODS: The animal model of bone defect was made on the bilateral radius of 54 New Zealand white rabbits, which were randomly divided into the experimental group(the defect was repaired with CHAP-FS-SAI), control group(with autograft) and blank control group(the defect was left unrepaired) with 18 rabbits in each group. The ability of bone defect repair was evaluated by gross observation, histopathological study, X-ray and biomechanical analysis 2, 4, 8 and 12 weeks after repair. RESULTS: (1) In the 2nd week, tight fibro connection could be found between the implant and fracture site and there were many fibroblasts and capillary proliferation with many chondrocytes around CHAP in the experimental group, while only a few callus formed, and chondrocytes, osteoblast and osteoclast existed in the control group. (2) In experimental group and control group, a large quantity of callus was found 4 and 8 weeks; ossification of chondrocytes with weave bone formation were found 4 weeks and many osteocytes and weave bones and laminar bones were found 8 weeks. (3) In the 12th week, the complete ossification of implant with well bone remodeling, a large number of mature osteocytes and laminar were found in experimental group and control group, and CHAP still existed in the experimental group; the defect area filled with fibro-scar tissue and only many fibroblasts could be seen in blank control group. (4) X-ray findings were the following: In experimental and control groups, callus formation could be seen 2 weeks postoperatively, more callus formed 4 weeks, the bone defect area disappeared and CHAP scattered in the callus 8 weeks; the fracture line disappeared and medullary cavity became united (in control group); and in the 12th week, the cortex became continuous, the medullary cavity became united, and remodeling completed, while bone defect was not still united in blank control group. The maximal torque and torsional stiffness in the experimental group is higher than those in the control group 2 weeks (P < 0.05), but there was no significant difference (P > 0.05) between the two groups 4, 8, 12 weeks after repair. CONCLUSION: The compound of CHAP-FS-SAI has good biological compatibility, and it can be used for one kind of bone substitute material to repair the bone defect. PMID- 14551931 TI - [Operative treatment of displaced talar neck fractures with absorbable lag screw]. AB - OBJECTIVE: To study a new kind of operation for displaced talar neck fractures. METHODS: From April 1996 to March 2001, 9 talar neck fractures were treated by internal fixation of absorbable lag screw with a medial approach and cut of medial malleolus to expose the fractures. A non-weight-bearing below-knee cast was applied for 6 to 12 weeks after operation. Once union of the fracture site is apparent, the patient should remain non-weight bearing in a removable short-leg and keep exercise every day. RESULTS: All the patients received follow-up from 15 to 60 months with an average of 28 months. The fractures healed from 20 to 42 weeks. The excellent and good rate of function was 77.8% (7/9) according to American Orthopedic Foot and Ankle Society Score(AOFAS). One case had the complication of superficial infection of wound and skin edge necrosis after operation, which was Hawkins type III. Late complication included two cases of avascular necrosis(AVN). Among them, one AVN of Hawkins type II was caused by early weight-bearing five weeks after operation and gained the fair score. The other AVN of Hawkins type III was inefficient to conservative therapy and proceeded ankle fusion in the end. The AOFAS of the patient was bad. CONCLUSION: Treatment of talar neck fractures by internal fixation of absorbable lag screw with a medial approach is an ideal method. It can gain a satisfactory result by the operation, strict postoperative care and rehabilitation. PMID- 14551932 TI - [Ultrastructure of anterior cruciate ligament after transplantation]. AB - OBJECTIVE: To study the characteristics of, morphology histology and ultrastructure of anterior cruciate ligament(ACL) autograft and two-step cryopreserved ACL allograft after transplantation. METHODS: Sixty New Zealand rabbits and sixty Japanese rabbits were randomly divided into two groups: ACL autograft group and two-step cryopreserved ACL allograft group. Immunosuppressant were not used after transplantation. The histology and ultrastructure of the ACL of transplantation and normal knee were observed after 4 weeks and 12 weeks, respectively. RESULTS: The rate of remodeling process was faster in ACL autograft than in two-step cryopreserved ACL allograft, but there was similar remodeling process between two groups 12 weeks after transplantation. The proportions of large-diameter fibers(> or = 80 nm) of ACL autograft and cryopreserved ACL allograft were 6% and 24% in the 4th week, and were 0 and 2% in the 12th week, respectively. The proportions of small-diameter of fibers(< 80 nm) of ACL autogrft and cryopreserved ACL allograft were 94% and 76% in the 4th week, and 100% and 98% in the 12th week, respectively. Histologic incorporation in ACL autograft was similar to that in cryopreserved ACL allograft. CONCLUSION: Two step cryopreserved bone-ACL-bone allograft were similar to bone-ACL-bone autograft cryopreserved in remodeling process and histology. The rate of remodeling process was faster in ACL autograft than in cryopreserved ACL allograft. PMID- 14551933 TI - [Reconstruction of a cervical esophagus segment with an artificial prosthesis by use of a polyurethane stent covered with collagen-chitosan sponge in dogs]. AB - OBJECTIVE: To repair esophageal defects with an artificial prosthesis composed of biodegradable materials and nonbiodegradable materials, which is gradually replaced by host tissue. METHODS: The artificial esophagus was a two-layer tube consisting of a chitosan-collagen sponge and an inner polyurethane stent with a diameter of 20 mm and a length of 50 mm. We used the artificial esophagus to replace 5 cm esophageal defects in group I (five dogs) and in group II (ten dogs), and nutritional support was given after operation. The inner polyurethane stent was removed after 2 weeks in group I and after 4 weeks in group II endoscopically and epithelization of the regenerated esophagus was observed by histologic examination and transmission electron microscope. RESULTS: In group I, the polyurethane stent was removed after 2 weeks, and partial regeneration of esophageal epithelial was observed; and constriction of the regenerated esophagus progressed and the dogs became unable to swallow after 4 weeks. In group II, the polyurethane stent was removed after 4 weeks, highly regenerated esophageal tissue successfully replaced the defect and complete epithelization of the regenerated esophagus was observed. After 12 weeks, complete regeneration of esophageal mucosa structures, including mucosal smooth muscle and mucosal glands and partial regeneration of esophageal muscle tissue were observed. CONCLUSION: Esophageal high-order structures can be regenerated and provided a temporary stent and support by polyurethane stent and an adequate three-dimensional structure for 4 weeks by collagen-chitosan sponge. PMID- 14551934 TI - [Characteristics of P38 mitogen-activated protein kinase and c-Jun expression in hypertrophic scar and their effects on scar formation]. AB - OBJECTIVE: To observe the protein expression of phosphorylated form of P38 mitogen-activated protein kinase(P38MAPK) and c-Jun in hypertrophic scar skin and to explore their influences on the formation and maturation of hypertrophic scar. METHODS: The expression intensity and distribution of phosphorylated form of P38MAPK and c-Jun were examined with immunohistochemistry and pathological methods in 16 cases of hypertrophic scar skin and 8 cases of normal skin. RESULTS: In normal skin, the positive signals of phosphorylated form of P38MAPK mostly distributed in basal lamina cells of epidermis, while c-Jun was mainly located in epidermal cells and endothelial cells. The positive cellular rates of two proteins were 21.3% +/- 3.6% and 33.4% +/- 3.5% respectively. In proliferative hypertrophic scar skin, the particles of phosphorylated P38MAPK and c-Jun were mainly located in epidermal cells and some fibroblasts. The positive cellular rates of two proteins were significantly elevated to 69.5% +/- 3.3% and 59.6% +/- 4.3% respectively (P < 0.01). In mature hypertrophic scar, the expression of these proteins decreased but was still higher than that of normal skin. CONCLUSION: The formation and maturation of hypertrophic scar might be associated with the alteration of phosphorylated P38MAPK and c-Jun protein expression in hypertrophic scar. PMID- 14551935 TI - [Effects of nandrolone phenylpropionate on the expression level of pro alpha 1 (I) collagen after burn in rats]. AB - OBJECTIVE: To explore the effects of nandrolone phenylpropionate (NP) on the expression level of pro alpha 1 (I) collagen after burn in rats and the possible mechanism involved in the process. METHODS: Thirty-two Wistar rats with a deep second-degree scald injury and 20% of total body surface area were randomly divided into two groups to receive either 5 mg/kg NP(NP group) or normal saline (control group) every other day. We analyzed the mean integrated optical density(mIOD) of androgen receptor (AR) to determine the distribution and expression of AR in fibroblasts by immunohistochemistry, and measured expression level of pro alpha 1 (I) collagen mRNA by quantitative fluorescent RT-PCR to find the relation between expressions of AR and pro alpha 1 (I) collagen mRNA. The total specimens were obtained from the scalded rats after 4, 7, 14 and 21 of after burn. RESULTS: The expression of pro alpha 1 (I) collagen mRNA in NP group was significantly higher than that in control group on the 7th, 14th and 21st days(P < 0.05), but there was no significant difference on the 4th day. The density of AR in fibroblasts had significant difference (P < 0.05) between the two groups after 4, 7, 14 and 21 days. A positive relationship existed between the expression of pro alpha 1 (I) collagen mRNA and quantity of AR in fibroblasts(r = 0.836). CONCLUSION: The nandrolone phenylpropionate increased the expression of pro alpha 1 (I) collagen mRNA and enhanced the density of AR in fibroblasts. The higher expression of pro alpha 1 (I) collagen mRNA had a relation with the change of quantity of AR in fibroblasts. PMID- 14551937 TI - [Treatment of overtime avulsion of scalp with split thickness scalp skin grafting: 7 cases of reports]. AB - OBJECTIVE: To investigate a treatment method for overtime avulsion of scalp. METHODS: Form October 1992 to July 2001, we treated 7 cases of avulsed scalp, which had been wounded more than 12 hours and accompanied with shock and head wound, with split thickness scalp skin grafting. RESULTS: Except for partial necrosis of scalp in center of bare area of skull, more than 90% of grafting split thickness scalp skin survived in 4 cases and more than 80% in 3 cases, and presented satisfactory appearance during following up. The bare area had no periosteum above 4 cm in diameter needed to graft split thickness skin after skull was covered granulation tissue. CONCLUSION: The limits of time of scalp skin grafting will be prolonged as long as the processes are settled properly to maintain the skin of body. PMID- 14551936 TI - [Endothelial cell proliferation stimulated by basic fibroblast growth factor]. AB - OBJECTIVE: To determine the influence of basic fibroblast growth factor (bFGF) on endothelial cell (EC) proliferation in vitro and its possible mechanisms, and to examine the effect of both TNP-470 and dexamethasone (Dex) on the EC proliferation induced by bFGF. METHODS: Human umbilical vein endothelial cells were cultured and the proliferation of EC was quantified by a colorimetric assay using MTT reagent. The expression of nuclear factor-kappa B (NF-kappa B) and ki 67 was detected with SABC immunohistochemical method. RESULTS: bFGF stimulated the EC proliferation and enhanced the expression of NF-kappa B and ki-67 in nucleus; TNP-470 and Dex suppressed EC proliferation induced by bFGF, and reduced the expression of NF-kappa B and ki-67 in nucleus. CONCLUSION: The above results indicate that the possible mechanisms of EC proliferation stimulated by bFGF come from that bFGF can activate NF-kappa B to promote the synthesis of DNA and EC mitosis. TNP-470 and Dex inhibited EC proliferation stimulated by bFGF by inhibiting NF-kappa B. PMID- 14551938 TI - [Identification of ectomesenchymal stem cells of human fetal facial processes and spontaneous differentiation to smooth muscle cells]. AB - OBJECTIVE: To investigate the characteristic and phenotype of ectomesenchymal stem cells of human fetal facial processes and the procedure of spontaneous differentiation to smooth muscle cells. METHODS: The primary ectomesenchymal cells of E 50 human fetal facial processes were isolated by 2.5 g/L trypsin and cultured with DMEM/F 12 with 10(-6) U/L leukemia inhibitor factor(LIF). The morphology and growth rate were observed by inverted microscop. After being withdrawn LIF, the characteristic of cells were identified by immunohistochemistry and RT-PCR. Ultrastructure was observed by transmission electron microscope. RESULTS: The cultured cells displayed monolayer growth and were fibroblast-like with 2-4 processes. The cells were stainely positived for anti-human natural killer cell marker-1, Vimentin, S-100, neuron specific enolase, myoglobin and VIII factor, but negatively for glial fibrillary acidic protein, neural fiblament, alpha-SMA and cytokeratin in immunohistochemistry. Two days after being withdrawn the LIF, cells expressed alpha-SMA in protein and mRNA levels. The cells were rich in muscular filament-like structure and dense bodies under transmission electron microscope. CONCLUSION: Cultured cells are undifferentiated ectomesenchymal stem cells. The cells have the potential for differentiating spontaneously to smooth muscle cell. PMID- 14551939 TI - [Effect of electric stimulation on denervated skeletal muscle atrophy]. AB - OBJECTIVE: To study the influence of the electric stimulation of denervated muscle atrophy. METHODS: Sixteen SD rats were made the model of denervated skeletal muscle in two lower limbs by cutting off the sciatic nerve and femoral nerve. The right gastrocnemius muscle was stimulated with JNR-II nerve & muscle recovery instrument by skin as the experimental side and the left was not treated as the control side. The muscle histology, ultrastructure, fibrillation potential amplitude, Na(+)-K(+)-ATPase and Ca(2+)-ATPase activities were observed 2 weeks and 4 weeks after operation. RESULTS: Electric stimulation could protect mitochondria and sarcoplasmic reticulum from the degeneration. The reduction rates of muscle cell diameter and cross section in the experimental side were slower significantly than those in control side. There was no influence on fibrillation potential amplitude in the both sides after electric stimulation. The reduction rates of Na(+)-K(+)-ATPase activity in the experimental side were slower 15.59% and 27.38% respectively than those in the control side. The reduction rates of Ca(2+)-ATPase activity in the experimental side were slower 4.83% and 21.64% respectively than those in the control side. CONCLUSION: The electric stimulation can protect muscle histology, electrophysiology and enzymic histochemistry of denervated skeletal muscle from the degeneration. The electric stimulation is an effective method to prevent and treat muscle atrophy. PMID- 14551940 TI - [Early microsurgical treatment of upper obstetrical brachial plexus injury]. AB - OBJECTIVE: To search for the operation timing and methods for obstetrical brachial plexus injury (OBPI). METHODS: Thirty-two children with upper OBPI were treated by microsurgical procedure from October 1997 to April 2001. The average age of patients was 10 months, ranged from 3 months to 24 months; of them, 19 were below 6 months while 13 were over 6 months. Surgical procedure included neurolysis(n = 12), coaptation after resection of the neuroma without function (n = 7), phrenic nerve transfer to anterior cord of upper trunk or musculocutaneous nerve (n = 7) and intercostal nerves transfer to musculocutaneous nerve(n = 6). The children underwent operation with microsurgical technique and 7/0 or 9/0 nylon was used for nerve suture. RESULTS: Thirty cases were followed up for 21 months postoperatively; the excellent and good rate was 76.7% (23/30). The results of the children under 6 months were better than those over 6 months. CONCLUSION: The microsurgical operation might be considered at the age of 3-6 month infants who had shown little or no improvement in elbow flexion. Neurolysis and nerve coaptation are superior to neurotization. The appropriate procedure should be selected according to the findings of exploration. PMID- 14551941 TI - [Allosclera transplant for eyelid reconstruction]. AB - OBJECTIVE: To study the clinical effects of allosclera transplant for eyelid reconstruction. METHODS: From May 1986 to January 2001, 45 patients who suffered from partial eyelid defect underwent the allosclera transplant for eyelid reconstruction, There were 31 cases of male and 14 cases of female, 18 cases of the upper eyelid and 27 cases of the lower eyelid, among them, 22 cases were caused by eyelid trauma and 23 cases by eyelid tumor. The length of eyelid defect were less than 1/2 of the whole eyelid. The morphology and function of the eyelid and complications were observed after operation. RESULTS: All 45 cases were followed up for 11 to 38 months, 19.7 months on average. The eyelid morphology and mobility restored well and there was no influence on visual function. CONCLUSION: The xenosclera is an ideal material for eyelid reconstruction. PMID- 14551942 TI - [Repair of segmental bone defects in rhesus monkeys' radius with allogeneic tissue engineered bones]. AB - OBJECTIVE: To construct tissue engineering bone with bio-derived materials and bone marrow stromal cells (MSCs), and to investigate the effect of allogeneic engineering bone implants on healing of segmental bone defects. METHODS: MSCs being aspirated aseptically from tibial tuberosities of young rhesus monkeys were induced into osteoblasts in vitro and then were cultured and marked with 5-bromo 2-deoxyuridine (BrdU). Tissue engineering bones were constructed with these labeled osteoblasts being seeded onto bio-derived materials made from fresh human bones which were treated physically and chemically, Then the constructs were implanted in 15 allogeneic monkeys to bridge 2.5 cm segmental bone defects of left radius as experimental groups, bio-derived materials only were implanted to bridge same size defects of right radius as control group. and, 2.5 cm segmental bone defects of both sides of radius were left empty in two rhesus monkeys as blank group. Every 3 monkeys were sacrificed in the 1st, 2nd, 3rd, 6th and 12th weeks postoperatively and both sides of the implants samples were examined macroscopically, histologicaly, and immunohistochemicaly. The two monkeys in blank group were sacrificed in the 12th week postoperatively. RESULTS: Apparent inflammatory reactions were seen around both sides of the implants samples in the 1st, 2nd, 3rd weeks, but it weakened in the 6th week and disappeared at the 12th week. The labeled osteoblasts existed at the 6th week but disappeared at the 12th week. The bone defects in experimental group were repaired and the new bone formed in multipoint way, and osteoid tissue, cartilage, woven bone and lamellar bone occurred earlier when compared with control group in which the bone defects were repaired in 'creep substitution' way. The bone defects in blank group remained same size at the 12th week. CONCLUSIONS: Engineering bones constructed with bio-derived materials and MSCs were capable of repairing segmental bone defects in allogeneic monkeys beyond 'creep substitution' way and making it healed earlier. Bio-derived materials being constituted with allogeneic MSCs may be a good option in construction of bone tissue engineering. PMID- 14551944 TI - [Urethral epithelium culture by using L929 cells as trophoderm in vitro]. AB - OBJECTIVE: To study the technique and method of urethral epithelium culture in vitro, so as to lay the groundwork for reconstructing a tissue engineering urethra and to provide an experimental model of urethral mucosa in physiological, pathological, toxicological and microbiological study. METHODS: The urethral mucosa from a young male New Zealand hare that had just been out of milk, was digested into single cell liquid with Dispase II and mixed enzyme, and the fibroblast were removed. After being seeded, the cells were cultured by using L929 cells as trophoderm. The medium was changed regularly and the cells were subcultured when they grew to mix together 80% to 90%. The cultured cells were analyzed with histochemistry, immunohistochemistry dyeing and flow cytometry examination. We observed the ultrastructure of cells with scanning electron microscope and transmission electron microscope. RESULTS: The primary cultured cells fused when they had been cultured for about ten days. They were the same in size like road rocks. The cultured cells were all epithelial cells without fibroblasts and were diploid cells. The cells could be subcultured 11-13 generations, and could survive 50-60 days. CONCLUSION: The urethral epithelium of young New Zealand hare can be cultured in vitro and maintain the ability to proliferate within a certain time. The study result not only sets a role in reconstructing a tissue engineering urethral mucosa, but also provides an experimental model for the research of urethral mucosa in vitro. PMID- 14551943 TI - [Effect of human telomerase reverse transcriptase mRNA introduction on the growth of human umbilicus vein endothelial cell]. AB - OBJECTIVE: To elongate the proliferation life-span of human umbilicus vein endothelial cell (HUVEC). METHODS: We synthesized the human telomerase reverse transcriptase mRNA (hTERT mRNA) by in vitro transcription, then transferred the hTERT mRNA into HUVEC in quicent stage by lipofect introduction. RESULTS: Telomerase expressed transiently in HUVEC, and the cell life-span was elongated for 7 population doublings. CONCLUSION: Telomerase can be reconstructed controllably and transiently in HUVEC by hTERT mRNA introduction, this method has the potential to be used to elongate the lifespan of cells cultured in vitro. PMID- 14551945 TI - [Apoptosis of fetus chondrocytes cultured in vitro and expression of caspase-3]. AB - OBJECTIVE: To investigate apoptosis of chondrocytes cultured in vitro and related expression of caspase-3. METHODS: Apoptosis of chondrocytes were detected by flow cytometry analysis and TUNEL staining. The expression of caspase-3 was determined by RT-PCR and Western blot, and caspase-3 protein activity was determined by ELISA. RESULTS: Apoptosis was observed in chondrocytes cultured in vitro from passage 1 to passage 4 at various degrees. The percentage of apoptosis of chondrocytes on day 7 was much higher than that on day 3 (15.7% +/- 0.3% vs 8.9% +/- 0.6%, P < 0.01). caspase-3 mRNA and protein expressed in chondrocytes during whole culture process. Along with the culture time extension in vitro, caspase-3 expression and protein activity up-regulated, coincident with apoptosis of chondrocyte. caspase-3 was activated and a fragment of 20 kDa was detected after 7 days of culture. CONCLUSION: caspase-3 is involved in apoptosis of chondrocytes cultured in vitro. PMID- 14551947 TI - [Preparation and characteristics of small intestinal submucosa]. AB - OBJECTIVE: To review the research advance of the preparation and characteristics of small intestinal submucosa(SIS). METHODS: Recent original articles related to such aspects of small intestinal submucosa were reviewed extensively. RESULTS: Small intestinal submucosa was an easily obtained biomaterial. SIS was a bio absorbable and degradable material. SIS had tissue specific regeneration properties. CONCLUSION: SIS is a suitable bio-derived material for tissue engineering of blood vessel, muscle tendon, urinary bladder and abdomen. PMID- 14551946 TI - [Component analysis of bone matrix via cell extraction]. AB - OBJECTIVE: To investigate the preparation of bone acellular extra-cell matrix(AECM) and to analyze its component. METHODS: With low-osmosis theory and method of cell extraction by detergent, bone acellular extra-cell matrix was prepared. We observed morphologic changes with HE, Mallory-Heidenhain rapid one step dyeing and Alcian blue dyeing and examined fibronectin(FN) and laminin(LN) with immunohistochemistry. RESULTS: Light microscope showed that the collagen fibers arranged regularly in AECM with blankness of bone lacunas by HE, Mallory Heidenhain rapid one-step dyeing and that the region around bone lacunas was stained different degrees of blue-green by Alcian blue dyeing. The result of immunohistochemistry showed there are positive markers of FN and LN in ECM. CONCLUSION: This method for preparation of bone acellular extra-cell matrix is effective, and it can keep natural structure of collagen fibers and maintain components of ECM, such as proteoglycan, FN and LN. PMID- 14551948 TI - [Preparation of ethanol-diesel fuel blends and exhausts emission characteristics in diesel engine]. AB - The technology that diesel oil is partly substituted by ethanol can reduce diesel engine exhausts emission, especially fuel soot. This research is concentrated on preparation of ethanol-diesel blend fuel and exhausts emission characteristics using diesel engine bench. Absolute ethanol can dissolve into diesel fuel at an arbitrary ratio. However, a trace of water (0.2%) addition can lead to the phase separation of blends. Organic additive synthesized during this research can develop the ability of resistance to water and maintain the stability of ethanol diesel-trace amounts of water system. The effects of 10%, 20%, and 30% ethanol diesel fuel blends on exhausts emission, were compared with that of diesel fuel in direct injection (DI) diesel engine. The optimum ethanol percentage for ethanol-diesel fuel blends was 20%. Using 20% ethanol-diesel fuel blend with 2% additive of the total volume, bench diesel engine showed a large amount decrease of exhaust gas, e.g. 55% of Bosch smoke number, 70% of HC emission, and 45% of CO emission at 13 kW and 1540 r/min. Without the addition of additive, the blend of ethanol produced new organic compounds such as ethanol and acetaldehyde in tail gas. However, the addition of additive obviously reduced the emission of ethanol and acetaldehyde. PMID- 14551949 TI - [Integrated effect of incorporation with different organic manures on CH4 and N2O emissions from rice paddy]. AB - Field experiment was conducted in 2001 rice growing season to understand the effect of organic manure application on CH4 and N2O emissions. Five treatments of fertilizer including urea and NH4HCO3 used as chemical fertilizer (CF), chemical fertilizer + pig manure (CF + PM), chemical fertilizer + cow manure (CF + CM), chemical fertilizer + rapeseed cake (CF + RC) and chemical fertilizer + wheat straw (CF + WS) were dedicated to this experiment. Results from the field observations indicated that application of the organic manure did not change the seasonal pattern of CH4 and N2O emissions but significantly affected the seasonal amounts. Seasonal amount of CH4 emission from the treatments of CF + CM, CF + RC and CF + WS was respectively 45%, 252% and 250% higher than that from the treatment of CF, but the application of pig manure (CF + PM) did not significantly influence the CH4 emission. The N2O emissions, on the other hand, were reduced by 18%, 21% and 18% for the treatments of CF + PM, CF + CM and CF + WS, respectively. Incorporation of rapeseed cake enhanced the N2O emission, yielding an increment of 22%. On a time scale of 20-year period, the integrated global warming potential (GWP) of CH4 + N2O released during the rice growing season for the treatments of CF + RC, CF + WS and CF + CM was respectively 154%, 136% and 17% higher than that for the CF treatment, while a reduction of 10% was obtained for the CF + PM treatment. The calculation of the GWP per unit rice grain yield suggested that the application of pig manure took the slight advantages not only in reducing the GWP but also in improving the grain yield. PMID- 14551950 TI - [Characteristics of atmospheric CO2 variations and some affecting factors in urban area of Beijing]. AB - The diurnal variation of the atmospheric CO2 at the urban area in Beijing displayed a very clear cycle, generally with the lowest value being recorded at about 15:00 local time and the highest in the nighttime. The differences between the maximums and the minimums ranged from 23.2 to 39.0 mumol.mol-1 in the four seasons and the ones of summer and fall were bigger than that of winter and spring. It showed a large seasonal cycle with a maximum in winter, from 421.5 to 441.0 mumol.mol-1, and a minimum in summer, between 367.4 and 371.6 mumol.mol-1. The peak-to-peak amplitude of atmospheric CO2 in Beijing, controlled mostly by the anthropogenic activities and seasonal variation of vegetation, was higher than that in Xinglong Station, Hebei Province and China Global Atmosphere Watch Baseline Observatory (Waliguan). The concentration of atmospheric CO2 in Beijing increased rapidly at a mean growth rate of 3.7%.a-1 from 1993 to 1995. After displaying a peak of 409.7 +/- 25.9 mumol.mol-1 in 1995, it decreased slowly. PMID- 14551951 TI - [Effect of translating water on vegetation at the lower reaches of Tarim river]. AB - Due to human irrational resource exploit in recent 50 years, many environment problems have appeared at the lower reaches of Tarim River, the environment degradation of this region became seriously. In order to restore green corridor and improve the environment at the lower reaches of Tarim River, the project of transporting water to the lower reaches of Tarim River was carried out from June 14, 2000 and the water reached Taitema lake finally. After two years' monitoring and analyzing in this region, it was found that the ground-water level went up and much extremely degraded vegetation was restored. Some xerophytic arbor and bush restored obviously with the change of habitat. Correspondingly, the groundwater level, vegetation coverage, diversity of biology, the species richness and crown of Populus euphratica all changed. By the end of September 2002, the result showed that the respond range of the ground-water level was 800 m from the river, and the respond range of vegetation was not beyond 700 m at the middle part of lower reaches of Tarim River. Due to the different environmental factors in different parts at the lower reaches of Tarim River, the response range changed accordingly. In general, the respond range became smaller and smaller from the upriver part to downstream part. PMID- 14551952 TI - [Influences of grazing and exclosure on carbon sequestration in degraded sandy grassland, Inner Mongolia, north China]. AB - Very little is known about the effects of continuous grazing and exclosure on carbon sequestration in semi-arid sandy grassland of North China. Soil-plant system carbon in representative degraded sandy grassland in Horqin sandy steppe (N42 degrees 58', E 120 degrees 42') was measured. Three situations were compared: continuous grazing (CG), exclosure for 5 years (5EX) and exclosure for 10 years (10EX). Ground cover increased from the CG (35%) to the 5EX (63%) and to the 10EX (81%), and accordingly soil organic carbon and total plant components carbon increased from the CG (493 g.m-2 and 95 g.m-2) to the 5EX (524 g.m-2 and 146 g.m-2) and to the 10EX (584 g.m-2 and 309 g.m-2). In terms of the distribution of carbon in plant-soil system, 65.3%, 78.2% and 83.9% of plant-soil carbon was in soils at 0-15 cm in the 10EX site, in the 5EX site and in the CG site, respectively. The results suggested that continuous grazing in the erosion prone sandy grassland was very detrimental to vegetation, soil and surrounding environment. Under exclosure conditions, vegetation restoration and litter accumulation not only effectively protected soil from loss through wind erosion, but also significantly increased plant-soil system carbon storage, and thus sequestration of atmospheric carbon. However, the recycling of aboveground plant carbon to the soil was restricted when grazing was excluded and a large part of carbon sequestrated was immobilized in plant litter accumulating on the soil surface. Further research on time scale for exclosure is needed. It is concluded that the degraded sandy grassland could contribute to significant carbon sequestration with the implementation of protecting practices. PMID- 14551953 TI - [Spatial heterogeneity of Ulmus pumila open forest ecosystem in Otindag sandy land]. AB - Ulmus pumila open forest ecosystem plays an important role in Otindag Sandy Land. Using biostatistics (U test and Pearson correlation analysis) and geostatistics (Semivariogram analysis and Kriging interpolation), the spatial heterogeneity of soil organic matter (SOM), soil total nitrogen (STN), soil pH (SpH), soil water content (SWC), and the correlations between them, as well as their relationships with herbaceous coverage (HC) and Ulmus pumila were studied. The results showed that spatial heterogeneity and spatial dependence were apparent in SOM, STN, SpH, SWC and HC. With Kriging interpolation in the sampling area, the contour maps for these properties were drawn. Using the contour maps, the relationship between Ulmus pumila and spatial heterogeneity of the studied properties was investigated. The possible reason for the formation of fertility island under the Ulmus pumila trees in the ecosystem was discussed. PMID- 14551954 TI - [Effect of inorganic nitrogen compounds and pH on the growth of duckweed]. AB - The use of duckweed in domestic wastewater treatment was paied increasing attention recently. But inorganic nitrogen compounds and pH of wastewater possibly affect the growth of duckweed. The effect of ammonia, nitrate and pH on the growth of Lemna minor L., a common spiece in China, was assessed with laboratory scale experiments. The experiments used artificial culture with different levels of pH, ammonia and nitrate concentrations. The experimental results indicated that the lowest value of pH Lemna minor can tolerate was between 5-6, and it can grow well in pH from 6 to 9. The growth rate of Lemna minor was inhibited gradually with increasing concentration of ammonia. The toxicity of ammonia was a result of both forms, NH3 and NH4+. Compared to NH4+, the effect of NH3 was much stronger. Nitrate had few inhibitory on the growth of Lemna minor. The increase in ammonia and nitrate concentrations can increase the chloropyll content of Lemna minor. Activity of peroxidase of Lemna minor in ammonia culture was higher than that in nitrate culture because of the toxicity of ammonia. PMID- 14551955 TI - [Enhance of reproduction of Phanerochaete chrysosporium by adding natural lixiviums in liquid medium]. AB - Great promotion to the reproduction of white rot fungus (Phanerochaete chrysosporium) by adding natural lixiviums such as from wood, maize core and potato in liquid medium was found in this research. Incubated in the liquid medium contained 10 g/L glucose as carbon source with natural lixiviums for three days, the production of mycelium pellet reached more than 80 g/L, which was 5 times of without natural lixiviums. Incubated in the liquid medium contained 5 mg/L glucose as carbon source with natural lixiviums for three days, the production of mycelium pellet can reach 69.5 g/L, while the production of medium without natural lixiviums was very low. When the basic medium contained 1 g/L-20 g/L glucose as carbon source, the production of mycelium pellet in 3 days can only reach 12.5 g/L to 14.5 g/L. The media with potato lixiviums were easily contaminated and the media with maize core lixiviums were easily bulking, while the media with wood lixiviums were not easily contaminated and bulking. Medium with wood lixiviums can produce more pellet than other medium, endure contamination and keep better sedimentation capacity. So, wood lixivium is better additive to the culture of white rot fungi in liquid medium. Addition of the mixture of wood, maize core and potato lixiviums is of advantage to the production of mycelium pellet. The difference of the production in the media with different amount of wood lixiviums showed little in the first 3 days, while it expanded after 3 days. Wood lixiviums stimulate the growth of P. chrysosporium instead of supply organics which fungi need. PMID- 14551956 TI - [Effects of La on the growth of kidney bean seedling under Cd stress]. AB - In this study, kidney bean (Phaseolus vulgaris) was used as experiment-material, and effects of lanthanum on growth and metabolism of kidney bean seedling under cadmium stress was studied by water culture. The results showed that kidney bean seedling was chronically harmed by 30 mumol.L-1 Cd2+, however, its height and main root were reduced about 31.1% and 39.2%, meanwhile, leaf area, fresh and dry weight of leaf, stem and root were decreased 48.0%, 42.7%, 29.6%, 61.3% and 49.4% respectively. With acute damage of 50 mumol.L-1 Cd2+, physiological and biochemical characteristics of seedling changed greatly, e.g., chlorophyll content and root activity were decreased about 23.5% and 28.7%, cell membrane permeability, the malonydialdehyde (MDA) content, activities of catalase (CAT) and peroxidase (POD) were increased 5.58%, 28.6%, 0.6% and 7.0% respectively. And with longer treatment time, the damage became more serious. When 10 mg.L-1 La was used to spray on kidney bean seedling under cadmium stress one time, growth and metabolism of seedling was obviously reconditioned and the damage of Cd pollution was reduced. It is proved, by the experiment, that La can improve chlorophyll content(30.0%) and reduce cell membrane permeability (0.87%) and content of MDA(9.5%), and keep the activities of CAT (0.1%) and POD(1.6%) of kidney bean seedling. PMID- 14551957 TI - [EXAFS studies on adsorption-desorption mechanism of Zn at delta-MnO2-water interface]. AB - Microscopic structures of Zn(II) surface complexes adsorbed at a delta-MnO2-water interface were studied using extended X-ray absorption fine structure (EXAFS) spectroscopy. In a 0.1 mol/L NaNO3 solution of pH 5.50, Zn(II) was adsorbed onto the solid surface in the form of octahedral hydrous Zn(II) ions. The octahedral Zn(II) was linked to the structural unit of octahedral MnO6 of the delta-MnO2 surface by sharing the O atoms. The average bond length of RZn-O was (2.071 +/- 0.007)A (n = 3) and the Zn-Mn atomic distance was (3.528 +/- 0.006) A (n = 3), which corresponded to a corner-sharing linkage adsorption mode (weaker adsorption). Macroscopic adsorption-desorption isotherm experiments showed that, in contrast to that of Zn-manganite, adsorption of Zn(II) on delta-MnO2 was highly reversible and no apparent adsorption hysteresis was observed. EXAFS results indicated that the microscopic mechanism for the high adsorption reversibility was corresponded to the weak adsorption sites of corner sharing linkage between the adsorbate and adsorbent polyhedra. PMID- 14551958 TI - [Analysis of factors resulting in anomalous arsenic concentration in groundwaters of Shanyin, Shanxi province]. AB - The chemical characteristics of groundwaters containing anomalous arsenic were summarized and the factors influencing arsenic enrichment in groundwater were discussed in this paper, based on chemical data of general compositions, trace elements and organic matters of sixty-six groundwaters and mineral analysis of thirty aquifer matrix. The main factors inducing anomalous arsenic in groundwaters included high pH (8.09), high concentrations of phosphate (0.71 mg/L) and organic matters (5.14 mg/L), and anoxic environment. High pH, high phosphate content and anoxic environment made against adsorption of As into aquifer which contains clay minerals. And high concentration of organic matters promoted the activity of As in groundwater system. These factors contribute to desorption and transportation of As in aquifers. PMID- 14551959 TI - [Bulking controls induced by nitrogen insufficiency in yeast system]. AB - The effect of nitrogen addition to salad oil manufacturing wastewater system was studied. The results of batch experiments demonstrated: 1. addition of nitrogen was helpful to improve oil removal, and the oil removal rate was above 90% when COD/N value was between 50/1 and 20/1; 2. addition of nitrogen improved the sludge settleability, which was much better under COD/N value of 50/1 and 20/1. In the continuous experiment, the value of slude volume index(SVI) was between 100 and 200, and the removal rates of oil and COD were above 95% and 90%, respectively when the COD/N ratio was kept at value of 20/1. Nitrogen addition to salad oil manufacturing wastewater system is an effective method for preventing or controlling sludge bulking. PMID- 14551960 TI - [Speciation and characterization of interaction of PACls with sulfate]. AB - The interaction of sulfate with various PACl was investigated by using Ferron assay, chemical analysis and SEM. The experimental results showed that the basicity (B = [OH]/[Al]) exhibited significant role in the PACl-sulfate reaction. It indicated different species in various PACl under different reaction pathway with sulfate. The Alc formed precipitation quickly with sulfate, while Alb underwent slowly crystalization. The decrease of Ala resulted in the limit of ferron method. The different speciation component in PACls formed different crystal morphology and chemical composition with sulfate. Increase the basicity, the content of sulfate in precipitate decreased from 0.45 to 0.30. PMID- 14551961 TI - [Removal of phthalate esters from drinking water using ozone-GAC process]. AB - This study investigated the feasibility of using ozone-GAC process to remove phthalate esters from drinking water through a batch-scale study and adsorption isotherms. Dimethyl phthalate (DMP), diethyl phthalate (DEP) and dibutyl phthalate (DBP) were selected as the representative of phthalate esters. Results indicated that ozonation remove more than 40% DMP, DEP and DBP, GAC absorbed all the DMP, DEP and DBP that had not been oxidized by ozone at the condition of the Empty Bed Contact Time (EBCT) from 4 minutes to 12 minutes. The isotherms for GAC were successful correlated by Freundlich equation, and the date was used to estimate GAC service time. The results indicated that ozone-GAC process is a feasible way to remove DMP, DEP and DBP from drinking water. PMID- 14551962 TI - [Relation between phosphorus and bacterial regrowth in drinking water]. AB - In the experiment, a bioassay called Bacterial Regrowth Potential (BRP) was used to investigate the effect of phosphorus on bacterial regrowth in the water sample that was made from some raw water taken from a reservoir located in northern China. It was shown that BRP of water sample increased about 100%-235% when 50 micrograms/L PO4(3-) -P (as NaH2PO4) was added alone. BRP of water sample with various inorganic nutrients (including phosphate) addition had the similar increase compared with BRP of water sample with only 50 micrograms/L PO4(3-) -P addition and BRP of water sample with acetate-C of 1 mg/L addition increased only 30%-40%. These results were clear evidence of phosphorus limitation on bacteria regrowth in the water sample investigated in the experiment. This observation offered novel possibility to restrict microbial regrowth in water distribution system by developing technologies to remove phosphorus efficiently from drinking water. PMID- 14551963 TI - [Influence of external electric current on the spatial distributions of porosity and fractional dimension of biofilms]. AB - The spatial distributions of porosity and fractional dimension of autotrophic and heterotrophic/autotrophic biofilms applied by direct current in the range of 0-25 A/m2 were investigated by using the micro-slicing technique and image analysis. The results showed that the porosities of the autotrophic and heterotrophic/autotrophic biofilms respectively decreased from ca. 92% and 96% in the top layer to ca. 54% in the bottom layer while the fractional dimensions increased from 1.1 and 1.05 in the top layer to 1.4 in the bottom layer in the absence of current. The increase in the current density decreased the porosities and increased the fractional dimensions in the top layers. In the deeper layers, the current had insignificant influence on the porosity and fractional dimension. PMID- 14551964 TI - [Organic acids conversion in methanogenic-phase reactor of the two-phase anaerobic process]. AB - Experiments on organic acids conversion in methanogenic-phase of the two-phase anaerobic process were conducted. The results showed that when the acidogenic phase was in mixed acids fermentation and its load was 41.5 kg/(m3-d), the load of methanogenic-phase was 6.05 kg/(m3.d), the substrate conversion velocity of bacteria inhabited at the same height of UASB reactor as follows: acetic acid > ethanol > butyric acid > propionic acid; ethanol-utilized microbial had the same pH range as methanogenic bacteria; ethanol-type fermentation was the optimal acidogenci-type fermentation for the two-phase anaerobic process; the conversion of acetic acid was rather high, but the others organic acids will be convert to acetic acid make it the rate-limiting step for anaerobic degradation. PMID- 14551965 TI - [Cultivation of aerobic granular sludge for simultaneous nitrification and denitrification in SBR system]. AB - The cultivation of aerobic granular sludge used for nitrogen removal in Sequencing Batch Reactor (SBR) fed with a synthetic municipal wastewater was investigated. The results showed that the aerobic granular sludge with the high activity of simultaneous nitrification and denitrification, could form in the reactor by controlling the addition of carbon source, and the removal rates of COD and NH3-N reached 74.0%-92.8% and 82.3%-98.5%, respectively, in the reactor. The granular formed was averagely 0.5-1.0 mm in diameter, and the Mixed Liquid Suspended Solids (MLSS) and the Sludge Volumetric Index (SVI) in the reactor were above 4.5 g/L and above 32.5, respectively. Compared with the normal aerobic activated sludge process, the system with aerobic granular sludge had a higher concentration of effective biomass and a much stronger ability of nitrogen removal. PMID- 14551966 TI - [Studies on aerobic granular sludge cultivated under selective pressure]. AB - As inoculum sludge from a conventional COD removal activated sludge wastewater treatment plant was used, aerobic granular sludge was cultured in sequencing batch reactor (SBR) fed with glucose as carbon substrate. The SBR was operated with decreasing sedimentation time and increasing COD loading rate resulting in the selective pressure to enhance the formation of granular sludge. According to morphology change of sludge, the generation of granule could be divided into three stage. Granules started to appear after 67 days operation. After increasing COD loading rate to 4.8 kg/(m3.d), and superficial gas velocity to 0.0175 m/s, granules were the dominant sludge forms with diameter about 6-9 mm, the minimal settling velocity of 32.7 m/h, and the MLSS of 7800 mg/L. Aerobic granular sludge demonstrates high activity and ability to withstand high COD loading rate. The properties of aerobic granular sludges and the effect of different operational conditions on the formation of these aerobic granules were also analyzed preliminarily. PMID- 14551967 TI - [Phosphorus removal of agriculture wastewater through subsurface constructed wetland]. AB - Phosphorus removal in subsurface constructed wetland treating agriculture non pointed wastewater was studied in pilot-scale. The experimental results showed that the removal rates of TP increased with HRT increasing when the nominal hydraulic retention time was less than 5 days. The pH decreased distinctly when the retention time was too longer in the batch and sequence style, it may bring on a lower removel rate of TP. The amount of TP that can be removed by harvesting were about 15.8% and 9.5% compared to the total quantity of removing in Phragmitas communis and zizania caduciflora constructed system respectively. Temperature impacted on the removel of TP, the removel rate of TP decreased 30% in the winter compared to summuer. PMID- 14551968 TI - [Chemical denitrification of nitrate from groundwater]. AB - Batch experiments for catalytic reduction of nitrate from groundwater with Pd and/Cu catalysts were conducted. It was found that Pd-Cu combined catalysts at a ratio of 4 can maximize the nitrate reduction into nitrogen; above 80% total nitrogen removal efficiency was realized in this study. It was also found that the catalytic activity was affected by the amounts of catalysts, hydrogen flow rate and pressure, the initial concentration of nitrate. With an increasing in the amount of the catalysts, both nitrite and ammonia intermediates can be kept at a low level. A high flow rate and pressure of hydrogen was in favor of the catalytic reduction of nitrate, but not benefit for the selectivity. The catalytic reduction followed a first order kinetics in term of the initial nitrate concentration. PMID- 14551969 TI - [Photocatalysis characterization of titanium dioxide supported on natural clinoplilolite]. AB - This paper studied preparing photocatalyst supported on natural clinoplilolite, photocatalysis degrading to methyl orange solution as photocatalysis function test, solar as light resource, explored the synthesize condition and affect factors of its catalysis activity. The capability of catalyst was evaluated by decolor rate and COD removal rate. The samples was described by XRD, IR and specific surface area. Studied result showed that catalyst prepared by combination of tetrabutyl titanate and natural clinoplilolite dryed under 120 degrees C for 6 hours then calcined under 200 degrees C had the best photocatalysis activity. Degrading rate of methyl orange solution increased with the quantity of TiO2/zeolite and additional oxidant H2O2 increasing, but superfluous H2O2 can also restrain the photocatalysis activity of titanium dioxide supported on clinoplilolite. Methyl orange solution had the best degrading rate as pH value between 2 to 5. PMID- 14551971 TI - [Effect of extraneous rare earths (REs) on form of soil exchangeable REs and heavy metals]. AB - The effects of extraneous rare earths (REs) on the form of exchangeable REs and heavy metals Fe, Mn and Zn in yellow cinnamon soil in different terms were studied through soil culture in both dry and inundated condition. The result showed that the treatments of REs enhanced the contents of soil exchangeable REs (EX-REs) and its distribution coefficients also increased while most still remained under 10%. The contents of EX-REs in inundated condition were higher than that in dry condition, but they decreased with the duration of soil culture. The composition of EX-REs in soil gradually approached that in extraneous REs, and it remained relatively steady over the treatment with 200 mg.kg-1 applied REs. Applying REs raised the contents of exchangeable heavy metals, and the contents of exchangeable Fe, Mn and Zn increased with linear trend, of which the effect of REs on Mn was the strongest one. PMID- 14551970 TI - [Characteristics of water-gas two-phase CO2 transport in unsaturated zone]. AB - The migration of CO2 in soil was simulated through use of soil column, especially the transformation of CO2 between water and gas phases at wetting front was studied. The soil column in the test was 1 m long and the concentration of CO2 solution was 748 mg/L. Through analysis, it was shown that CO2 migration in water phase was governed by advection, dispersion, reaction and mass transfer between water and gas phases, that in gas phase by advection, dispersion, mass transfer between two phases. Sampling method under dynamic condition of two phase flow in soil column was approached and mass transfer coefficient from water phase to gas phase was determined as 0.00061. This showed that there was certain mass transformation of CO2 from water phase to gas phase and yet it was not so significant. This study can provide the quantitative scientific bases for safety assessment of geological disposal of low and intermediate level radioactive waste and environmental impact assessment of solid waste disposal by landfilling. PMID- 14551972 TI - [Distribution of fluoride in the combustion products of coal]. AB - The static distribution characteristic of fluoride in the combustion products of coal was studied by ashing procedure of coal, and the dynamic distribution characteristics of fluorine in the combustion products of coal in pulverized-coal fired boiler and layer-burning boiler were investigated. Experimental results identified that fluorine in coal belong to volatile elements, fluorine in fly ash and bottom ash were non-rich. About 94.5% of the fluorine in coal emitted as gaseous-fluorine during coal combustion in pulverized-coal-fired boiler, and about 80% of the fluorine in coal emitted as gaseous-fluorine during coal combustion in layer-burning boiler. 55%-60% of the fluorine in fly ash of pulverized-coal-fired boiler were distributed in fly ash particles with a diameter of 74 microns-104 microns. PMID- 14551973 TI - [The effect of microstructure on the desulphurization of absorbents]. AB - The microstructure and the pore size distribution of 3 kinds of shell and 1 kind of limestone were measured with electronic scanning microscope and pore master mercury porosimeter, and the desulphurization characteristics of above absorbents were studied with thermal-gravimetric analyzer. As a result, the grains of calcium from shell like tortuous flakes, with pore diameter above 0.2 micron and higher pore volume, while the grains from limestone like particles, with pore diameter between 0.01 to 0.16 micron and lower pore volume, but the surface area were very high. The bigger pore which was above 0.2 micron in shell had many advantages, such as fewer air-blocked pores, lower gas diffusing resistance, almost simultaneous and completed reaction. So, finally the calcium conversation ratio of the shell was greater than that of the limestone. On the other hand, while the gas diffusion was meted, it was effective for desulphurization to reduce the pore size and to increase the surface area. PMID- 14551974 TI - [Experimental study on DC corona radical shower for the removal of toluene]. AB - Using DC corona radicals shower to decompose toluene in air, different parameters were studied, such as the concentration of toluene, temperature, voltage, humidity and the settled time. The results showed that the fall of temperature and appropriate humidity can increase the decomposing efficiency of toluene; the efficiency decreased with the increasing of the voltage; the settled time was longer, the decomposing efficiency was higher, but the energy efficiency decreased. Increasing the concentration of toluene made the removal efficiency lower, but in the same time, made the energy efficiency higher. The experiment built a good basis for researching radicals decompose PAHs(dioxins etc.). PMID- 14551975 TI - [Evaluation of Fenton process for the treatment of sanitary landfill leachate with Response Surface Methodology]. AB - Response Surface Methodology (RSM) was employed to investigate the effect of TOC ratio of high molecular weight organic matter (HMWOM) to low molecular weight organic matter (LMWOM), the LMWOM concentration and hydrogen peroxide to ferrous ion molar ratio on COD removal during the treatment of sanitary landfill leachate with Fenton reagents. The high and low levels of HMWOM/LMWOM, the LMWOM concentration and H2O2/Fe2+ were from 14 to 38, 975 to 3000 mg/L, and 1.75 to 4.00 respectively. Using the software of Design--Expert 5, a response surface quadratic model in terms of actual factors was obtained based on the experimental data, and COD removal contour plots were derived from the model accordingly. It showed that COD removal increased with the increase of HMWOM to LMWOM ratio, but decreased with increasing LMWOM concentration. There existed an optimal hydrogen peroxide to ferrous ion molar ratio so that the highest COD removal efficiency could be achieved. PMID- 14551976 TI - [Accumulation of heavy metals in the sediments of Shenzhen Bay, south China]. AB - Heavy metals concentrations in marine sediment cores of Shenzhen Bay were measured, and the profile distribution characteristic of heavy metals was discussed. Combined with the 210 Pb dating results, the contamination history of heavy metals was studied in high resolution records, and the metal accumulation processes were also analyzed. The results indicated that the concentrations of heavy metals was relatively low compared with other area in the world, but the elements of Pb, Cu and Zn were obviously contaminated by anthropogenic impact. The rapid economical development of Shenzhen in the last 20 years and Hong Kong in 1960-1970s contributed much on accumulation of heavy metals in the sediments. PMID- 14551977 TI - [Study on activated carbon regeneration by heterogeneous catalytic wet oxidation]. AB - The CuO-Al2O3 catalyzer was used to heterogeneous catalytic wet oxidation regeneration of activated carbon, which adsorbed the phenol saturated. The study found the effect of reaction factors on activated carbon regeneration by heterogeneous catalytic wet oxidation. And the optimum reaction condition was reaction temperature 210 degrees C, reaction time 60 min, catalyzer dosage 25 mg (calculated by Cu), reaction oxygen pressure 0.6 MPa at 25 degrees C, carbon 15 g (dry carbon), distilled water 300 mL. It was found that the catalyzer had good stability in this experiment by using X-rey radiation analysing catalyzer and experiment on catalyzer stability. PMID- 14551978 TI - [The computer simulating prediction method of noise of underground garage in urban residential region]. AB - The model of random point sound source was applied to predict the noise of underground garage in urban residential region with the method of computer simulating. Comparing the predict data and the data of measurement, the precision of this model was good. The model can be applied to predict the influence of the noise of new built underground garage noise, it can offer the reference for the planning department. PMID- 14551979 TI - [The disappearance and transfer behavior of PCBs bestrewed into semi-field soil systems]. AB - The disappearance and transfer behavior of polychlorinated biphenyl (PCBs) (the mains of penta-PCBs) at higher concentrations (400 mg/kg) bestrewed into semi field soil systems were studied under near field conditions (semidry climate in North China). The result showed that PCBs did not lose and transfer forward below after 12 months of treatment. And after 24 months of treatment, only less than 2% of PCBs transferred vertically no more than 10 cm and PCBs concentration at treatment layers decreased 10.19% including the amount of transfer. It provides the scientific foundation for the landfill or other treatment measurements of PCBs, also gives the basic data for ecological risk assessment of PCBs and other relative compounds. PMID- 14551980 TI - Relevant environmental science. PMID- 14551981 TI - Comment on general fugacity-based model for multiple chemical species. PMID- 14551982 TI - Time-dependent toxicokinetics of [14C]lindane in the terrestrial isopod Porcellionides pruinosus. AB - Preliminary acute soil toxicity tests are a starting point for risk assessment. These tests are performed at one exposure time and are used to estimate the lethal concentration for 50% of the population (LC50). However, it is known that LC50 varies over time, following an exponential decay model. Here, we present the variation of LC50 over time in the isopod Porcellionides pruinosus exposed to [14C]lindane when considering the concentrations measured in bulk soil and in extracted water. Using a wide range of concentrations, the percent mortality was recorded over various time intervals. Higher concentrations strongly influenced isopod survival. The LC50 infinity value for P. pruinosus was 3.57 micrograms/g for bulk soil concentration, suggesting a great sensitivity of this species to lindane. Simultaneously, the values estimated for the lethal body concentration (LBC) were 2.36 micrograms/g animal for bulk soil concentrations and 2.79 micrograms/g animal when extracted water concentrations are considered. An alternative to the LC50 determination is the estimation of LBC, which is proposed as a better way to describe the acute toxicity of chemicals. Kinetic-based toxicity models were fitted to the data and revealed uptake rate constants of 1.1 g soil/g animal/week and 84.3 ml extracted water/g animal/week for bulk soil and extracted water concentrations, respectively. Elimination rate constants of 1.7 per week were found for both pathways of exposure. PMID- 14551983 TI - Identification of toxic products of anthracene photomodification in simulated sunlight. AB - Currently, the evidence of a rapid photomodification of anthracene under sunlight resulting in enhanced toxicity exists; however, the chemical causes of toxicity are still unknown. The present study aimed at filling this gap by irradiation of an anthracene suspension with simulated sunlight and subsequent effect-directed fractionation and analysis of toxic products with respect to the inhibition of bacterial energy metabolism of Vibrio fischeri, reproduction of the green algae Scenedesmus vacuolatus, and genotoxicity in the umuC test. Algal toxicity of anthracene was hardly modified by irradiation prior to testing and distributed over all fractions with emphasis on the fractions containing anthracene-9,10 dione and a photometabolite suggested to be 10-hydroxyanthrone. Bacterial toxicity and genotoxicity in contrast emerged only when anthracene was irradiated. Anthracene-1,4-dione, a so-far-unknown trace photometabolite, was identified as a very potent toxicant dominating the toxicity of photomodified anthracene to V. fischeri. In genotoxic fractions, 1-hydroxyanthracene-9,10-dione and 1,4-dihydroxyanthracene-9,10-dione were identified and confirmed as genotoxicants. The results stress the potential of effect-directed analysis approaches in contrast to mere chemical analysis in studies aiming at toxicologically relevant photomodified substances. PMID- 14551984 TI - Bacterial detection of the toxicity of dioxins, polychlorinated diphenyls, and polybrominated diphenyl ethers. AB - Polychlorinated dibenzo-p-dioxins (PCDDs), biphenyls (PCBs), and polybrominated diphenyl ethers (PBDEs) were found to induce several specific stresses within bacterial cells. Four different recombinant Escherichia coli with specific stress promoters (i.e., the recA responsive to DNA damage, fabA responsive to membrane damage, katG responsive to oxidative damage, and grpE responsive to protein damage) that were fused to the lux operon from Vibrio fischeri showed very unique specificities in terms of their stress responses in the presence of PCDD and PCBs. In addition, a recombinant bacterium with the lac promoter fused to the lux operon from Xenorhabdus luminescence also showed dose-dependent responses via a loss of bioluminescence because of the addition of the PCDDs and PCBs. Brominated diphenyl ethers (BDE) -47, -99, and -153 congeners, however, were not found to induce any stress within the bacterial cells, indicating that these chemicals do not stimulate any cellular stresses related to those tested. These three congeners, however, did result in different levels of general cellular toxicity, which was found to be dependent on the position of the bromine. Finally, the cellular toxicity within the bacteria was found to increase when exposed to mixtures of dioxins, PCBs, and PBDEs, possibly from synergistic effects. PMID- 14551985 TI - A toxicity identification evaluation approach to studying estrogenic substances in hog manure and agricultural runoff. AB - Spreading liquid manure on agricultural fields is a routine way of disposing of animal manure and optimizing the use of nutrients for crops. Limited studies suggest that these wastes may contain a variety of endocrine-disrupting compounds (EDCs) that may be released into aquatic environments through runoff. The purpose of this study was to apply a toxicity identification and evaluation approach to isolate and identify estrogenic compounds in hog manure. A recombinant yeast estrogen screen bioassay was used to detect estrogenicity of high-performance liquid chromatography--separated hog manure fractions. Further analytical analyses of the fractions and comparison to authentic standards resulted in the identification of the endogenous estrogens 17 beta-estradiol (E2) and estrone, and the phytoestrogen metabolite, equol. High levels of equol (6.9-16.6 ppm) were found to be present in manure that was stored for several months. The endocrine disrupting potential of equol was characterized further by using fish hormone estrogen receptor (ER), sex hormone binding protein (SSBP), and goldfish androgen receptor (AR) radioligand binding assays. Equol was found to be approximately 1,000- and 200-fold less potent that E2 in competing for binding sites of the SSBP and ER, respectively. Equol's potency was 2,200-fold less than testosterone for the AR. Additional studies confirmed the presence of compounds with estrogenic activity in tile drain water after application of hog manure to an agriculture field. In this case, the contribution of equol to the total estrogenicity of the tile drain water was minimal relative to that of natural estrogens. Overall, this study indicates that further work is warranted to assess the impact that EDCs that originate from agricultural runoff may have on the ecology or physiology of exposed biota. PMID- 14551986 TI - Quantitative structure-biodegradation relationships for ortho-substituted biphenyl compounds oxidized by Methylosinus trichosporium OB3b. AB - Methanotrophs, bacteria that thrive in the presence of stable methane and oxygen concentrations, can cometabolically oxidize ortho-substituted biphenyls to yield a variety of hydroxylated products. Despite awareness of the susceptibility of ortho-substituted biphenyls and other aromatic compounds to methanotrophic oxidation, the molecular properties relevant for predicting rates of methanotrophic oxidation are unknown. To this end, we have developed quantitative structure-biodegradation relationships using oxygen uptake activity by the type 2 methanotroph. Methylosinus trichosporium OB3b, expressing the soluble form of methane monooxygenase and in the presence of nine ortho-substituted biphenyls. Multivariate analysis yielded the strongest correlations using the initial slope of the oxygen uptake rate versus substrate concentration curve as the dependent variable. Quantum mechanical descriptors, including the sum of carbon charges on the substituted ring, the charge on the substituted carbon, and the width of compound calculated using computationally derived bond lengths and dihedral angles, correlated more strongly with oxygen uptake activity than did empirically derived electronic descriptors. The resulting models suggest a significant influence of substituent electronic nature and size and the involvement of the substituted carbon site in the oxidation of these compounds by M. trichosporium OB3b. PMID- 14551988 TI - Sublethal effects of copper on coho salmon: impacts on nonoverlapping receptor pathways in the peripheral olfactory nervous system. AB - The sublethal effects of copper on the sensory physiology of juvenile coho salmon (Oncorhynchus kisutch) were evaluated. In vivo field potential recordings from the olfactory epithelium (electro-olfactograms) were used to measure the impacts of copper on the responses of olfactory receptor neurons to natural odorants (L serine and taurocholic acid) and an odorant mixture (L-arginine, L-aspartic acid, L-leucine, and L-serine) over a range of stimulus concentrations. Increases in copper impaired the neurophysiological response to all odorants within 10 min of exposure. The inhibitory effects of copper (1.0-20.0 micrograms/L) were dose dependent and they were not influenced by water hardness. Toxicity thresholds for the different receptor pathways were determined by using the benchmark dose method and found to be similar (a 2.3-3.0 micrograms/L increase in total dissolved copper over background). Collectively, examination of these data indicates that copper is broadly toxic to the salmon olfactory nervous system. Consequently, short-term influxes of copper to surface waters may interfere with olfactory-mediated behaviors that are critical for the survival and migratory success of wild salmonids. PMID- 14551987 TI - Tenax extraction mimics benthic and terrestrial bioavailability of organic compounds. AB - Biota to sediment accumulation factors (BSAFs) are widely used to describe the potential accumulation of organic contaminants in organisms. From field studies it is known that these BSAFs can vary dramatically between sediments of different origin, which is possibly explained by the variation in bioavailability of organic contaminants in sediments. In the present study it is shown that the variability in BSAF values for different sediment samples obtained at two Dutch freshwater sites could largely be explained by the variation in Tenax-extractable concentrations in these sediments. Variations of a factor of about 50 could be explained. The ratio between concentrations in biota and Tenax-extractable concentrations in sediment varied slightly between sediments and contaminant class, but was close to the theoretically expected value of 2. This is a strong indication that Tenax-extractable concentrations of contaminants in sediments are an excellent indicator of available concentrations. PMID- 14551990 TI - Comparison of synthetic surfactants and biosurfactants in enhancing biodegradation of polycyclic aromatic hydrocarbons. AB - Polycyclic aromatic hydrocarbon (PAH) contamination of the environment represents a serious threat to the health of humans and ecosystems. Given the human health effects of PAHs, effective and cost-competitive remediation technologies are required. Bioremediation has shown promise as a potentially effective and low cost treatment option, but concerns about the slow process rate and bioavailability limitations have hampered more widespread use of this technology. An option to enhance the bioavailability of PAHs is to add surfactants directly to soil in situ or ex situ in bioreactors. Surfactants increase the apparent solubility and desorption rate of the PAH to the aqueous phase. However, the results with some synthetic surfactants have shown that surfactant addition can actually inhibit PAH biodegradation via toxic interactions, stimulation of surfactant degraders, or sequestration of PAHs into surfactant micelles. Biosurfactants have been shown to have many of the positive effects of synthetic surfactants but without the drawbacks. They are biodegradable and nontoxic, and many biosurfactants do not produce true micelles, thus facilitating direct transfer of the surfactant-associated PAH to bacteria. The results with biosurfactants to date are promising, but further research to elucidate surfactant-PAH interactions in aqueous environments is needed to lead to predictive, mechanistic models of biosurfactant-enhanced PAH bioavailability and thus better bioremediation design. PMID- 14551989 TI - Synthesis and estrogenic activity of bisphenol a mono- and di-beta-D glucopyranosides, plant metabolites of bisphenol A. AB - The syntheses and characterization of bisphenol A mono- and di-beta-D glucopyranosides were undertaken to confirm that these compounds are major plant metabolities of bisphenol A (BPA) and to allow an assessment of their estrogenicity. Synthesis involved the glucosidation of unprotected BPA with glucose penta-acetate with phosphorus oxychloride as catalyst. The estrogenic activity of BPA and its mono- and di-beta-D-glucopyranosides were measured with an enzyme-linked immunosorbent assay (ELISA)-based estrogen receptor competitive binding assay and with a yeast two-hybrid assay adapted to a chemiluminescent reporter gene (for beta-galactosidase). Both methods showed that the estrogenicity of BPA was eliminated by formation of the di-glucoside, but whereas the ELISA-based method indicated that reduced activity remained in the monoglucoside, the yeast two-hybrid method showed the monoglucoside to be inactive. Presumably these results reflect the more complex interactions of test compound and cellular components required to demonstrate estrogenicity in the yeast two-hybrid assay. As these processes parallel those in mammalian cells, the yeast two-hybrid method is likely to be the more realistic assay. The uptake and metabolism of BPA by plants offers the possibility of phytoremediation of contaminated water, but also provides an additional route for the compound to enter the human food chain. PMID- 14551991 TI - Mutagenicity of nitroaromatic degradation compounds. AB - The mutagenicity of 2,4-dinitrotoluene (24DNT), and 2,6-dinitrotoluene (26DNT), and their related transformation products such as hydroxylamine and amine derivatives, which are formed by Clostridium acetobutylicum, were tested in crude cell extracts using Salmonella typhimurium TA100. A previous publication already reported the mutagenic activities of 2,4,6-trinitrotoluene (TNT) and its related hydroxylamine derivatives in this test system. A time course of the mutagenicity during the anaerobic transformation of TNT, 24DNT, and 26DNT was also investigated under the same conditions to compare with the results from the pure compounds. The monohydroxylamino intermediates 2-hydroxylamino-4-nitrotoluene (2HA4NT), 4-hydroxylamino-2-nitrotoluene (4HA2NT) and 2-hydroxylamino-6 nitrotoluene (2HA6NT) formed during anaerobic transformation of dinitrotoluenes were proven to be mutagenic in the Ames test using Salmonella typhimurium TA100. This study reports that 4HA2NT is the most stable derivative, whereas 2HA4NT and 2HA6NT are less stable and these intermediates are mutagenic in the Ames test. Both 24DNT and 26DNT and their final metabolites 2,4-diaminotoluene (24DAT) and 2,6-aminotoluene (26DAT) appeared nonmutagenic. In a time-course study of TNT degradation, the temporal sample containing 85% of 2,4-dihydroxylamino-6 nitrotoluene (24HA6NT) is most mutagenic. These observations suggest that the bioremediation approach for treatment of 24DNT and 26DNT should be carried past the hydroxylamino intermediate. PMID- 14551992 TI - Linear free-energy relationship analysis of the fate of chlorinated 1- and 2 carbon compounds by redox-manipulated smectite clay minerals. AB - This paper expands on earlier work on the effect of molecular structure on the fate of a series of chlorinated 1- and 2-carbon compounds (RCl), including carbon tetrachloride, 1,1,1-trichloroethane, 1,1,2,2-tetrachloroethane, pentachloroethane, hexachloroethane, trichloroethene, tetrachloroethene, trichloronitromethane, and trichloroacetonitrile, in the presence of unaltered (SWa-U) and reduced (SWa-R) ferruginous smectite. A linear free-energy relationships (LFERs) analysis was conducted to explain kinetic data for RCl adsorption and dechlorination. Correlation analyses based on RCl hydration energies, hydrolysis rate constants, or bond dissociation energies were significant (r2 > 0.87) for RCl adsorption by SWa-R but not so for RCl adsorption by SWa-U (r2 < 0.5). The most statistically significant relationships describing RCl kinetic data were achieved using multivariable regression analysis including RCl hydration energies (or hydrolysis rate constant) and bond dissociation energies for RCl heterolysis. The outcome of this study indicates that the oxidation state of structural Fe in the clay strongly influences RCl hydration. A mechanism consistent with the present LFERs analysis corresponds to RCl hydration as the limiting step for adsorption and dechlorination by redox-manipulated smectites. PMID- 14551993 TI - Effect of sorption on benzene biodegradation in sandy soil. AB - The effect of sorption on benzene biodegradation in sandy soil was studied by conducting kinetic microcosm batch tests in soil-free solution and in the presence or absence of bacteria in soil materials with varying degrees of powdered activated carbon (PAC). In the soil-free experiment, benzene was added to a solution inoculated with Pseudomonas aeruginosa bacteria in order to achieve a potential or maximum biodegradation rate. In subsequent experiments, benzene was applied to a solution containing sandy soil and various PAC contents with and without inoculating P. aeruginosa. Benzene concentrations in the soil-free experiments decreased with time with two characteristic rates. A two-stage exponential decay model adequately represented the observed solution concentration pattern with time. Sorption experiments in bacteria-free soil also decreased monotonically, with the extent of sorption increasing as PAC content increased. The sorption data were represented well with a two-stage irreversible sorption model. A third set of experiments in the presence of both soil and bacteria showed more rapid concentration loss from solution than the set of experiments with bacteria-free soil. A model combining sorption and degradation greatly overestimated the loss when the rate coefficient from the bacteria-free experiments was used. Satisfactory agreement between model predictions and observed values was obtained when the degradation rate coefficients were decreased by factors ranging from 3 to 10, depending on the amount of PAC present. Model predictions of the percentage benzene mass remaining in the soil after 25 d of degradation ranged from 72 to 97%, depending on the PAC content, compared to only 2.5% remaining in soil-free solution. PMID- 14551994 TI - Mineralization of desorption-resistant 1,4-dichlorobenzene in wetland soils. AB - Laboratory studies were conducted to investigate the biologically mediated, aerobic mineralization of both freshly added and artificially aged, desorption resistant 1,4-dichlorobenzene (1,4-DCB). The adsorption and desorption of 1,4-DCB isotherms were established in three wetland soils using decant-refill batch techniques. Significant nonlinearity and hysteresis were observed in the isotherms with a hysteresis index ranging from 0.11 (relatively low hysteresis) in a marsh soil to 2.26 (relatively high hysteresis) in a bottomland hardwood soil from the Petro Processor (PPI) Superfund site. Mineralization of freshly added 1,4-DCB was observed in all three soils without lag after the addition of a 1,4-DCB degrading culture. Mineralization curves were plotted above theoretical lines predicted from a first-order model assuming instantaneous desorption, indicating that the microbial population had access to sorbed 1,4-DCB. In separate experiments, mineralization of artificially aged, desorption-resistant 1,4-DCB was also observed. Mineralization curves in these studies also indicated that the microbial population could directly access sorbed 1,4-DCB. The extent and rate of mineralization of desorption-resistant 1,4-DCB decreased significantly, including rate constants decreasing from approximately 0.01 d-1 in the freshly added treatments to approximately 0.002 d-1 in the desorption resistant treatments. Although sorption/desorption partitioning helped explain mineralization patterns in the treatments with freshly added 1,4-DCB, no differences were observed in mineralization curves in the desorption-resistant treatments between soils with widely varying sorption/desorption properties. PMID- 14551995 TI - Enhancing toxicity test performance by using a statistical criterion. AB - Aquatic toxicity tests are laboratory experiments that measure the biological effect (e.g., growth, survival, reproduction) of effluents, receiving waters, or storm water on aquatic organisms. These toxicity tests must be performed using the best laboratory practices, and every effort must be made to enhance repeatability of the test method. We evaluated the generated reference toxicant test data for insurance of a level of quality assurance for tests over time within a laboratory and among laboratories. We recommend the reporting and evaluation of the percent minimum significant difference (PMSD) value for all toxicity test results. The minimum significant difference (MSD) represents the smallest difference between the control mean and a treatment mean that leads to the statistical rejection of the null hypothesis (i.e., no toxicity) at each concentration of the toxicity test dilution series. The MSD provides an indication of within-test variability, and smaller values of MSD are associated with increased power to detect a toxic effect. We recommend upper and lower PMSD bounds for each test method in order to minimize within-test variability and increase statistical power. To ensure that PMSD does not exceed an upper bound, testing laboratories may need to increase replication, decrease variability among replicates, or increase the control mean performance. PMID- 14551997 TI - Expression and inducibility of aryl hydrocarbon receptor pathway genes in wild caught killifish (Fundulus heteroclitus) with different contaminant-exposure histories. AB - Wildcaught killifish from a contaminated site on the Elizabeth River (VA, USA) are refractory to induction of cytochrome P4501A (CYP1A, measured as catalytic activity and immunodetectable CYP1A protein) after exposure to typical aryl hydrocarbon receptor (AHR) agonists, as has been reported for fish from other sites highly contaminated with these compounds. In an attempt to understand the molecular basis for the lack of inducibility of CYP1A protein expression and activity in Elizabeth River killifish, we analyzed the expression of CYP1A and four other members of the AHR signal transduction pathway: AHR1, AHR2, AHR repressor (AHRR), and AHR nuclear translocator (ARNT). Gene expression was measured by cycle-optimized reverse transcriptase-polymerase chain reaction (RT PCR) analysis of messenger ribonucleic acid (mRNA) extracted from livers of killifish from the Elizabeth River and King's Creek (VA, USA) (reference site), 36 h after injection with beta-naphthoflavone (BNF, an AHR agonist) or corn oil (carrier control). Hepatic CYP1A mRNA was inducible in King's Creek killifish. However, in Elizabeth River killifish, no induction of CYP1A mRNA was observed, confirming and extending previous results showing no induction of CYP1A protein or catalytic activity in this population. Similarly, AHRR and AHR2 mRNA levels were induced by BNF in King's Creek but not Elizabeth River killifish. No population or treatment-related differences were observed in expression of AHR1 or ARNT mRNAs. The results reveal in Elizabeth River killifish a consistent lack of inducibility of genes that are normally inducible by AHR agonists (CYP1A, AHRR, AHR2). However, the expression of AHR1, AHR2, and AHRR in vehicle-treated fish did not differ between Elizabeth River and King's Creek killifish, suggesting that altered constitutive expression of AHRs or AHRR does not underlie the refractory CYP1A phenotype in Elizabeth River killifish. PMID- 14551996 TI - Determination of the estrogenic and antiestrogenic effects of environmental contaminants in chicken embryo hepatocyte cultures by quantitative-polymerase chain reaction. AB - A method was developed to measure the estrogenic and antiestrogenic effects of various chemicals and organochlorine extracts in chicken embryo primary hepatocyte cultures. Messenger RNAs (mRNAs) for the estrogen-inducible egg yolk proteins, vitellogenin II (VTGII), and very low-density lipoprotein apoprotein II (apoII), were measured by multiplex quantitative reverse transcription-polymerase chain reaction (Q-PCR). After 48 h of exposure, both VTGII and apoII mRNA levels were induced by moxestrol (1-1,000 nM), 17 beta-estradiol (10-1,000 nM), o,p'-DDT (apoII: 1,000 and 10,000 nM, VTGII: 10,000 nM), 4-tertoctylphenol ([OP]; apoII: 20 and 50 microM, VTGII: 10-50 microM), and methoxychlor ([MXCL]; apoII: 5-50 microM, VTGII: 20 and 50 microM). Tamoxifen (100 and 1,000 nM) induced apoII mRNA only, and bisphenol A (BPA) was not estrogenic. Inhibition of moxestrol-mediated VTGII or apoII mRNA induction by MXCL, o,p'-DDT and tamoxifen indicated that these chemicals were also antiestrogenic at concentrations similar to those which caused estrogenic responses. Organochlorine extracts prepared from herring gull embryo yolk sacs obtained from three Great Lakes sites and one Atlantic coast site (reference site) did not show any estrogenic activities. However, the same extracts from all three Great Lakes sites had antiestrogenic activities. These results indicate that wild birds may be susceptible to the estrogenic or antiestrogenic effects of environmental contaminants. PMID- 14551998 TI - Inherited resistance to arsenate toxicity in two populations of Lumbricus rubellus. AB - No unequivocal evidence exists of genetically inherited resistance to metals/metalloids in field populations of earthworms. We studied cocoon production in adult Lumbricus rubellus Hoffmeister collected from an abandoned arsenic and copper mine (Devon Great Consols, Devon, UK), and abandoned tungsten mine (Carrock Fell, Cumbria, UK) and an uncontaminated cultured population. The earthworms were kept in uncontaminated soil for nine weeks. From a total of 42 L. rubellus from each site, Devon Great Consols adults produced 301 cocoons, of which 42 were viable; Carrock Fell 60 cocoons, of which 11 were viable; and the reference population 101 cocoons, of which 62 were viable. The hatchlings were collected and stored at 4 degrees C at weekly intervals. After 12 weeks, all hatchlings were transferred to clean soil and maintained at 15 degrees C for 20 weeks until they showed evidence of a clitellum. In toxicity trials, F1 generation L. rubellus were exposed to 2,000 mg As/kg as sodium arsenate or 300 mg Cu/kg as copper chloride for 28 d. The F1 generation L. rubellus from Devon Great Consols mine demonstrated resistance to arsenate but not copper. All L. rubellus from Devon Great Consols kept in soil treated with sodium arsenate remained in good condition over the 28-d period but lost condition rapidly and suffered high mortality in soil treated with copper chloride. The control population suffered high mortality in soil treated with sodium arsenate and copper chloride. Previous work has shown that field-collected adults demonstrate resistance to both arsenate and Cu toxicity under these conditions. Thus, while arsenate resistance may be demonstrated in F1 generation L. rubellus from one of the contaminated sites, Cu resistance is not. The F1 adults and F2 cocoons did not have significantly higher levels of As than the control population, with no residual As tissue burden, suggesting that resistance to As in these populations may be inherited. PMID- 14551999 TI - Phospholipase A2 activity in three species of littoral freshwater rotifers exposed to several toxicants. AB - We analyzed three species of Lecane, a littoral rotifer, for susceptibility to six metals and four organic toxicants using a fluorometric assay based on inhibition of activity of the enzyme phospholipase A2. The metallic toxicants that we tested included Cd, Cr, Cu, Pb, Hg (as HgCl2), and Ti; the organic toxicants included benzene, ethyl acetate, toluene, and vinyl acetate. The three species differed greatly with respect to their susceptibility to the various toxicants. Lecane quadridentata, for example, was particularly sensitive to the four organic compounds (median effective concentration values [EC50] ranged from 6.6 x 10(-4)-0.987 mg/L). Lecane luna, in contrast, seemed particularly sensitive to metals (EC50 values ranged from 2 x 10(-6)-1.92 mg/L). Lecane hamata was relatively insensitive to organic solvents (EC50 values ranged from 4.25-126.5 mg/L). PMID- 14552000 TI - Uptake, depuration, and toxicity of dissolved and sediment-bound fluoranthene in the polychaete, Capitella sp. I. AB - Deposit-feeders may be important in facilitating the removal of organic contaminants from bulk sediment. The relationship between uptake route (water vs sediment), depuration and toxicity was investigated in the deposit-feeding polychaete Capitella species I. Individual worms were exposed either to dissolved (i.e., < 0.45 micron) fluoranthene (FLU) (water-only treatment [WO]), or both to dissolved and sediment-bound FLU (porewater and sediment [PWS] treatment). The porewater concentration of dissolved FLU in PWS was set approximately equal to the dissolved concentration in WO (0, 50 micrograms FLU l-1). Fluoranthene marginally affected growth in WO worms (6 d exposure) but not in PWS worms (5 d exposure). The final body-burdens in PWS and WO worms were 141.2 and 99.5 micrograms total FLU equivalents (g dry wt worm)-1, respectively. Sediment-bound FLU was calculated to contribute at least 30% and up to as much as 91% of the total amount taken up by PWS worms. Water-only treatment worms retained all of the FLU during the subsequent depuration (4 d), whereas PWS worms decreased both the weight-specific (t1/2 = 0.95 d) and the total-body burden (t1/2 = 7.8 d), suggesting both active excretion and dilution of FLU body burden as a result of growth. Thus, our results indicate that the sediment-associated pool is an important route of uptake in Capitella sp. I, and that such deposit feeders may be important for remobilization of sediment-associated contaminants, such as fluoranthene. PMID- 14552001 TI - Biotransformation of dissolved and sediment-bound fluoranthene in the polychaete, Capitella sp. I. AB - Polycyclic aromatic hydrocarbons (PAHs) are widely distributed in the marine environment and have physicochemical properties that make them likely to bioaccumulate. The main purpose of this study was to investigate fluoranthene (FLU) uptake, biotransformation and elimination in the deposit-feeding polychaete Capitella sp. I exposed via different routes. Individual worms were exposed to either dissolved FLU or to both dissolved and sediment-bound FLU. The 14C-FLU was used as a tracer to determine the amounts of unmetabolized FLU (parent), water soluble (aqueous)- and water-insoluble (polar) FLU metabolites and FLU residues (unextractable fraction) in sediment, water, and worm tissue. Capitella was capable of accumulating and biotransforming FLU regardless of route of exposure, thus suggesting that biotransformation activity is not restricted to gut tissues. Although both feeding and nonfeeding worms were able to biotransform FLU extensively, feeding worms eliminated/transformed parent FLU faster than both aqueous and polar FLU metabolites after transfer to clean conditions, whereas nonfeeding worms barely excreted FLU in any form. The high biotransformation capability of Capitella sp. I. along with the dependency of elimination on exposure route, demonstrates the potential importance of these processes in controlling the fate and effects of PAH and possibly other organic toxicants in contaminated marine sediments. PMID- 14552002 TI - Ecotoxicologic impacts of agricultural drain water in the Salinas River, California, USA. AB - The Salinas River is the largest of the three rivers that drain into the Monterey Bay National Marine Sanctuary in central California (USA). Large areas of this watershed are cultivated year-round in row crops, and previous laboratory studies have demonstrated that acute toxicity of agricultural drain water to Ceriodaphnia dubia is caused by the organophosphate (OP) pesticides chlorpyrifos and diazinon. We investigated chemical contamination and toxicity in waters and sediments in the river downstream of an agricultural drain water input. Ecological impacts of drain water were investigated by using bioassessments of macroinvertebrate community structure. Toxicity identification evaluations were used to characterize chemicals responsible for toxicity. Salinas River water downstream of the agricultural drain was acutely toxic to the cladoceran Ceriodaphnia dubia, and toxicity to C. dubia was highly correlated with combined toxic units (TUs) of chlorpyrifos and diazinon. Laboratory tests were used to demonstrate that sediments in this system were acutely toxic to the amphipod Hyalella azteca, a resident invertebrate. Toxicity identification evaluations (TIEs) conducted on sediment pore water suggested that toxicity to amphipods was due in part to OP pesticides; concentrations of chlorpyrifos in pore water sometimes exceeded the 10-d mean lethal concentration (LC50) for H. azteca. Potentiation of toxicity with addition of the metabolic inhibitor piperonyl butoxide suggested that sediment toxicity also was due to other non-metabolically activated compounds. Macroinvertebrate community structure was highly impacted downstream of the agricultural drain input, and a number of macroinvertebrate community metrics were negatively correlated with combined TUs of chlorpyrifos and diazinon, as well as turbidity associated with the drain water. Some macroinvertebrate metrics were also correlated with bank vegetation cover. This study suggests that pesticide pollution is the likely cause of ecological damage in the Salinas River, and this factor may interact with other stressors associated with agricultural drain water to impact the macroinvertebrate community in the system. PMID- 14552003 TI - Lethal and sublethal effects of atrazine, carbaryl, endosulfan, and octylphenol on the streamside salamander (Ambystoma barbouri). AB - Agricultural contaminants may be contributing to worldwide amphibian declines, but little is known about which agrichemicals pose the greatest threat to particular species. One reason for this is that tests of multiple contaminants under ecologically relevant conditions are rarely conducted concurrently. In this study, we examined the effects of 37-d exposure to the agrichemicals atrazine (4, 40, and 400 micrograms/L), carbaryl (0.5, 5, and 50 micrograms/L), endosulfan (0.1, 1, and 10 micrograms/L for 31 d and 0.1, 10, and 100 micrograms/L for the last 6 d), and octylphenol (5, 50, and 500 micrograms/L) and to a solvent control on streamside salamanders (Ambystoma barbouri) in the presence and absence of food. We found that none of the agrichemicals significantly affected embryo survival, but that hatching was delayed by the highest concentration of octylphenol. In contrast to embryos, larval survival was reduced by the highest concentrations of carbaryl, endosulfan, and octylphenol. Growth rates were lower in the highest concentrations of endosulfan and octylphenol than in all other treatments, and the highest concentration of endosulfan caused respiratory distress. Significantly more carbaryl, endosulfan, and octylphenol tanks had larvae with limb deformities than did control tanks. Refuge use was independent of chemical exposure, but 10 micrograms/L of endosulfan and 500 micrograms/L of octylphenol decreased larval activity. Systematically tapping tanks caused a greater activity increase in larvae exposed to 400 micrograms/L of atrazine and 10 micrograms/L of endosulfan relative to solvent controls, suggesting underlying nervous system malfunction. Hunger stimulated a decrease in refuge use and an increase in activity, but this response was least pronounced in larvae exposed to the highest concentration of any of the four agrichemicals, possibly because these larvae were the most lethargic. More studies are needed that concurrently examine the effect of multiple contaminants on amphibians so we can better identify effective mitigating measures. PMID- 14552004 TI - Formation of benzo[a]pyrene and 7,12-dimethylbenz[a]anthracene adducts in vascular endothelia of cytochrome P4501A-induced chicken embryos. AB - Polycyclic aromatic hydrocarbons (PAHs) are widespread in the environment and birds may be exposed to PAHs via diet, from preening feathers contaminated with oil, or through contamination of the eggshell during embryo development. In the present study, tissue distribution and the cell-specific binding of two labeled PAHs, benzo[a]pyrene ([3H]BaP) and 7,12-dimethylbenz[a]anthracene ([3H]DMBA), were examined in chicken embryos exposed in ovo to CYP1A inducers. Tape-section autoradiograms revealed high concentrations of radioactivity in the bile, liver, kidneys, heart, and leptomeninges. Light microscopy autoradiography of solvent extracted tissue slices showed a high and selective binding in endothelial cells in certain blood vessels in brain, heart, lung, and chest muscle. Binding was also observed in blood vessel endothelial cells in the chorioallantoic membrane (CAM), an extraembryonal tissue lining the eggshell. Endothelial binding was confirmed in CAM exposed in vitro, implying that tissue-binding metabolites were formed in situ. The CYP1A inhibitor ellipticine abolished bleeding in the target endothelial cells in CAM. It is thus concluded that blood vessel endothelia in various tissues in birds can bioactivate environmental contaminants and be targets for their toxicity. In view of its critical position beneath the shell, the CAM could be an important target for toxicants following external exposure in oviparous species. PMID- 14552005 TI - Modifications to the fathead minnow (Pimephales promelas) chronic test method to remove mortality due to pathogenic organisms. AB - Whole effluent toxicity testing is used to evaluate the discharge of materials that may be harmful to indigenous aquatic life. Unlike most environmental analyses, receiving water (the water body into which the effluent is discharged) often is used as dilution water in whole effluent toxicity tests to simulate the aquatic environment into which the effluent is introduced. In approximately 26% of whole effluent toxicity tests conducted by Wisconsin (USA) certified labs between 1988 and 1998, a pathogenic effect caused failure of the receiving water controls during the fathead minnow chronic test (i.e., > 20% mortality). We performed microbiological work to isolate pathogenic organisms from receiving waters, the fish, and their food. We found pathogenic organisms such as Flexibacter columnaris, Aeromonas hydrophila, and Flavobacter spp. to be ubiquitous and were not able to remove them from the test (e.g., through decontamination of the fish food and carefully following sterility procedures). To eliminate the pathogenic effect, we evaluated manipulations of the sample and the test method including filtering receiving water, irradiating receiving water, using older fish (48 h), using clean test beakers each day of the test, and using smaller test beakers (30 ml) with two fish per beaker. In samples demonstrating the pathogenic effect, most of these manipulations significantly reduced mortality. The use of smaller tests cups was significantly better at reducing the effect than all of the other sample and method manipulations. These results indicate that a simple method modification to the fathead minnow chronic test will improve test reliability when diluting effluents with receiving waters. PMID- 14552006 TI - Similarities in bioaccumulation patterns of polychlorinated dibenzo-p-dioxins and furans and polychlorinated diphenyl ethers in laboratory-exposed oligochaetes and semipermeable membrane devices and in field-collected chironomids. AB - Sediment and chironomid samples were collected from a river in the vicinity of and downstream from a closed chlorophenol production plant, and they were analyzed for polychlorinated dibenzo-p-dioxins and furans (PCDD/F) and polychlorinated diphenyl ethers (PCDE). Bioaccumulation of these compounds from the sediments by the oligochaeta, Lumbriculus variegatus, and semipermeable membrane devices (SPMDs) was studied in a 28-d bioaccumulation test. The sum concentrations of PCDD/Fs and PCDEs in sediments varied spatially, peaking at the site below the production plant, but the congener composition remained fairly constant throughout the river. The sum concentrations of chironomids, oligochaetes, and SPMDs all varied independently of sediment concentrations. Both invertebrates accumulated more PCDEs, whereas PCDD/Fs dominated in sediments and SPMDs. Despite the variable sum concentrations, similarities were observed in concentrations and compositions of individual congeners. A significant correlation was found between sediments and oligochaetes, and between oligochaetes and chironomids, for both PCDD/Fs and PCDEs. These results suggest that bioaccumulation of PCDD/Fs and PCDEs in invertebrates is nonselective. Furthermore, the laboratory bioaccumulation tests seem to yield relevant results. The SPMDs were found to give good predictions of the PCDD/F concentrations of sediments and oligochaetes. For PCDEs, the estimates were slightly poorer, but still useful for preliminary concentration assessments. PMID- 14552007 TI - Examination of reproductive endpoints in goldfish (Carassius auratus) exposed in situ to municipal sewage treatment plant effluent discharges in Michigan, USA. AB - Effects of representative mid-Michigan (USA) wastewater treatment plant (WWTP) effluents on the reproductive physiology of common goldfish (Carassius auratus) were assessed in situ by measuring plasma concentrations of vitellogenin (VTG), 17 beta-estradiol (E2), and testosterone (T), and evaluating gonad histology. Caged adult male and female goldfish were exposed for six weeks to WWTP effluents. One riverine site and one lacustrine site were included as references for comparison to WWTP sites. There was differential growth and gonadal development among locations, which confounded the interpretation of biomarker responses. A linear contrast model was developed by analysis of covariance, and adjusted values were developed for concentrations of VTG, E2, and T in the plasma of both male and female fish. In general, VTG concentrations were significantly less in male than in female goldfish. Most concentrations of VTG in male goldfish were less than the method detection limit. There were no significant differences in plasma VTG of either male or female goldfish among locations or between sites at WWTPs and reference sites. Concentrations of E2 in the plasma of female goldfish were similar among WWTP sites, all of which were less than in females at a pond reference location. Concentrations of E2 in the plasma of male goldfish were similar at all WWTP locations, except for one, where they were greater. No consistent trends in hormone concentrations or gonadal histology could be attributed to putative endocrine disrupter exposure in WWTP effluents. The results indicate that the risk for estrogen agonist exposure below these mid Michigan WWTPs is small. PMID- 14552008 TI - Occurrence of polychlorinated biphenyls and polybrominated diphenyl ethers in green mussels (Perna viridis) from Singapore, Southeast Asia. AB - The green mussel, Perna viridis, was used in this study to measure levels of polychlorinated biphenyls (PCBs) and, for the first time, polybrominated diphenyl ethers (PBDE) in the marine environment. Samples were collected from eight different locations in the coastal waters of Singapore between April and May 2002. Forty-one PCB and 21 PBDE congeners were quantified by gas chromatography mass spectrometry and were all positively detected in the mussel tissues. Total concentrations in green mussel tissues ranged from 6.1 to 82 ng/g and 2.0 to 38 ng/g on a dry-weight basis for PCBs and PBDEs, respectively. Such levels reflect the ubiquity of these persistent organic pollutants in a tropical marine environment. Principal component analysis was applied to the PCB data and revealed similarities in the congener composition of mussel tissues to that of the commercial PCB mixture, Aroclor 1254. The PBDE levels, to date, were approximately one order of magnitude greater than the upper concentrations reported for blue mussel (Mytilus edulis) tissues in Europe. At some sampling sites, the congener composition of PBDEs in P. viridis tissues indicated recent exposure to a commercial pentabrominated flame retardant. PMID- 14552009 TI - Effects of 4-nonylphenol on reproduction of Japanese medaka, Oryzias latipes. AB - We exposed mature Japanese medaka (Oryzias latipes) to 4-nonylphenol (4-NP) to elucidate its reproductive effects and estrogenic responses. For each treatment, eight breeding pairs were exposed for 3 weeks at measured 4-NP concentrations of 24.8, 50.9, 101, and 184 micrograms/L. Egg production of the pairs during the third week of exposure (15-21 d) was significantly decreased in the > or = 101 microgram/L treatment group, and fertility was significantly decreased in the 184 microgram/L group. Induction of testis-ova was observed in male fish at all 4-NP concentrations, whereas abnormality of spermatogenesis was found only in those receiving the 184-microgram/L treatment. The gonadosomatic index of the males in the 184-microgram/L group was also remarkably reduced. Hepatic vitellogenin (VTG) levels were significantly increased in both sexes receiving 4-NP treatments of > or = 50.9 micrograms/L. Overall, the lowest-observed effective concentrations (LOECs) of 4-NP for decreasing reproductive capacity (fecundity and fertility) and eliciting estrogenic responses (induction of testis-ova and VTG) were 101 and < or = 24.8 micrograms/L, respectively. The LOEC for reduced reproduction in medaka was at least four-fold greater than that required to elicit estogenic responses in male fish. Thus, we suggest that the estrogenic responses expressed in male fish are not responsible for the reduction in reproductive capacity. PMID- 14552010 TI - The effect of lead-contaminated soil from Canadian prairie skeet ranges on the neutral red retention assay and fecundity in the earthworm Eisenia fetida. AB - The sublethal effects of lead (Pb) on the earthworm Eisenia fetida were evaluated in the laboratory using freshly spiked soil and soil collected from Canadian prairie skeet ranges. After a four-week exposure to soil spiked with lead acetate, earthworm neutral red retention time (NRRT). soil Pb concentrations, and earthworm Pb body burdens were measured. Lysosomal NRRT was reduced in a concentration-dependent manner (p < 0.0001), and NRRT was negatively correlated with earthworm Pb body burdens (r = -0.80, p < 0.0001). To evaluate the effects of aged Pb, earthworms were exposed to soil from three skeet ranges, and responses were compared with three matched reference sites. After a four-week exposure, NRRT, growth, fecundity, soil total Pb levels, and earthworm Pb body burdens were measured. The potentially bioavailable fraction of Pb in these sites was measured using a Ca(NO3)2 extraction. Growth and fecundity did not differ significantly between any of the skeet ranges and their reference sites. However, NRRT was significantly reduced in all three ranges compared with their respective reference sites (p < 0.05), indicating that the neutral red retention assay (NRRA) may be useful for detecting toxicity and potential hazards at Pb contaminated sites. Lysosomal NRRT was negatively correlated with soil Ca(NO3)2 extractable Pb (r = -0.80, p < 0.0001) and soil total Pb (r = -0.73, p = 0.001). Lysosomal NRRT was negatively correlated (r = -0.67, p < 0.002) with earthworm Pb tissue levels. PMID- 14552012 TI - Gonadal differentiation in frogs exposed to estrogenic and antiestrogenic compounds. AB - Exposure of amphibians to endocrine disrupting compounds (EDCs) may alter differentiation of gonads, especially when exposures begin during early life stages. Gonadal differentiation was observed in leopard frogs (Rana pipiens) and wood frogs (Rana sylvatica) exposed as tadpoles to estrogenic (estradiol, ethinylestradiol, nonylphenol) and antiestrogenic compounds (an aromatase inhibitor, flavone, and an antiestrogen, ICI 182780). Exposure to all compounds at micrograms/L concentrations altered gonadal differentiation in some animals by inducing either complete feminization or an intersex condition, and altered testicular tubule morphology, increased germ cell maturation (vitellogenesis), and oocyte atresia. Comparisons between the two species indicate that R. pipiens are more susceptible to sex reversal and development of intersex gonads. However, R. sylvatica also showed alterations to testicular morphology, germ cell maturation, and ooctye atresia. These laboratory results indicate that amphibians could be susceptible to altered gonadal differentiation and development when exposed to estrogenic and antiestrogenic compounds in aquatic environments, such as those impacted by agricultural, industrial, and municipal runoff. PMID- 14552011 TI - Development and field validation of a predictive copper toxicity model for the green alga Pseudokirchneriella subcapitata. AB - In this study, the combined effects of pH, water hardness, and dissolved organic carbon (DOC) concentration and type on the chronic (72-h) effect of copper on growth inhibition of the green alga Pseudokirchneriella subcapitata were investigated. Natural dissolved organic matter (DOM) was collected at three sites in Belgium and The Netherlands using reverse osmosis. A full central composite test design was used for one DOM and a subset of the full design for the two other DOMs. For a total number of 35 toxicity tests performed, 72-h effect concentration resulting in 10% growth inhibition (EbC10s) ranged from 14.2 to 175.9 micrograms Cu/L (factor 12) and 72-h EbC50s from 26.9 to 506.8 micrograms Cu/L (factor 20). Statistical analysis demonstrated that DOC concentration, DOM type, and pH had a significant effect on copper toxicity; hardness did not affect toxicity at the levels tested. In general, an increase in pH resulted in increased toxicity, whereas an increase of the DOC concentration resulted in decreased copper toxicity. When expressed as dissolved copper, significant differences of toxicity reduction capacity were noted across the three DOM types tested (up to factor 2.5). When expressed as Cu2+ activity, effect levels were only significantly affected by pH; linear relationships were observed between pH and the logarithm of the effect concentrations expressed as free copper ion activity, that is, log(EbC50Cu2+) and log(EbC10Cu2+): (1) log(EbC50Cu2+)= - 1.431 pH + 2.050 (r2 = 0.95), and (2) log(EbC10cu2+) = -1.140 pH -0.812 (r2 = 0.91). A copper toxicity model was developed by linking these equations to the WHAM V geochemical speciation model. This model predicted 97% of the EbC50dissolved and EbC10dissolved values within a factor of two of the observed values. Further validation using toxicity test results that were obtained previously with copper spiked European surface waters demonstrated that for 81% of tested waters, effect concentrations were predicted within a factor of two of the observed. The developed model is considered to be an important step forward in accounting for copper bioavailability in natural systems. PMID- 14552013 TI - A model to understand the confounding effects of natural sediments in toxicity tests with Chironomus riparius. AB - Recently, we built a model to link feeding input with growth, emergence, and reproduction of the midge Chironomus riparius exposed to an artificial sandy sediment. This model is based on assumptions about both feeding behavior and use of energy. Here, we show how it can be used for toxicity tests with natural sediments to understand and model the influence of sediment characteristics. We measured growth, emergence, and reproduction of chironomids exposed in beakers to four unpolluted natural sediments and three feeding conditions (no feeding, 0.2 mg Tetramin/larva/d, and 1.4 mg Tetramin/larva/d) and compared the results with data obtained on our artificial sandy sediment. Sediment characteristics had lower influence on growth than feeding level, but their influence could not be neglected. First, we could distinguish between sandy sediments and other sediments. This difference resulted in a significant delay of about 18 h in the growth curves. Second, in case of food limitation, chironomids could use the organic materials in the sediment, provided that the C:N ratio of the sediment was less than 14. Our model proved to be able to incorporate those two phenomena. As for reproduction, we observed a better reproduction (measured in number of eggs per mass) for natural sediments than for artificial sediments. We showed that this difference could be due to the lipid content of the natural sediments. PMID- 14552014 TI - Enantiomer-specific biomagnification of alpha-hexachlorocyclohexane and selected chiral chlordane-related compounds within an Arctic marine food web. AB - Concentrations of achiral and chiral organochlorine contaminants (OCs), including hexachlorocyclohexane isomers (HCH), chlordane congeners (cis- and trans chlordane, cis- and trans-nonachlor, MC5, MC7, and U82), and related metabolites (oxychlordane [OXY] and heptachlor exo-epoxide [HEPX]), were quantified in seawater (100 L; n = 6) and biota from the coastal Beaufort-Chukchi Seas food web near Barrow (AK, USA). The biota included zooplankton (Calanus spp.; n = 5), fish species such as arctic cod (Boreogadus saida; n = 10), arctic char (Salvelinus alpinus; n = 3), and marine mammals including bowhead whales (Balaena mysticetus; liver: n = 23; blubber: n = 40), beluga whales (Delphinapterus leucas; blubber: n = 20), ringed seals (Phoca hispida; blubber: n = 20), and bearded seals (Erignathus barbatus; blubber: n = 7). The food web magnification factors (FWMFs) for HCHs and chlordane compounds ranged from 0.5 (gamma-HCH) to 6.5 (HEPX) and were expected based on known recalcitrance and biotransformation of OCs. The enantiomer fractions (EFs) of all chiral OCs were near racemic (EF = 0.50) in the seawater, zooplankton, and all fish analyzed. In contrast, the EFs for most OCs analyzed were nonracemic (EF # 0.50) in the marine mammals blubber (range: 0.09 0.79) because of enantiomer-specific biotransformation and (or) accumulation. However, EF values were not significantly correlated with isotopically determined trophic level. The EFs for all chiral OCs (except alpha-HCH) in bowhead whale liver closely approximated the values in zooplankton, suggesting that the accumulation of chiral OCs from prey into this cetacean is not enantiomer specific. However, the modification of EFs from bowhead liver to blubber suggests that this species has the ability to enantioselectively biotransform and accumulate several chiral OC compounds. PMID- 14552015 TI - Sorption of phenanthrene and atrazine by plant cuticular fractions. AB - Several studies have shown selective preservation of plant cuticular materials in soils. However, very little is known about their function as sorbents for the hydrophobic organic contaminants (HOCs) in the soil. In this study, we investigated the sorption and desorption of phenanthrene and atrazine by cuticular fractions of pepper (bulk, dewaxed, nonsaponifiable, and nonhydrolyzable) to better understand the sorptive activity of cuticular matter in soils. The bulk and dewaxed cuticles exhibited carbon-normalized distribution coefficients (Koc) for phenanthrene and atrazine in the range of that reported for soil humic substances, although both samples were rich in aliphatic structures. No hysteresis was observed in the desorption isotherms of either solute. The nonhydrolyzable residue exhibited a very high Koc value for atrazine, whereas the nonsaponifiable sample be exhibited the lowest Koc value for both sorbates. Based on solubility parameter data, it is suggested that the nonsponifiable sample be considered an intermediate between the physical and chemical mixture of pectin and cutan/lignin-like fractions, whereas the dewaxed cuticle is a chemical blending of cutin and pectin. The n-hexane-normalized sorption data suggest that the pepper cuticle can interact specifically with atrazine. This study leads to the conclusion that the contribution of aliphatic rich plant biopolymers to the sorption of HOCs can be significant because of their preservation and accumulation in soils. PMID- 14552016 TI - Dynamics of lipids and polychlorinated biphenyls in a Baltic amphipod (Monoporeia affinis): a field study. AB - Processes such as accumulation and elimination, which control tissue concentration of polychlorinated biphenyls (PCBs), were examined over time in an in situ study of the amphipod Monoporeia affinis. These processes were studied with respect to individual PCB congeners, percentage lipid and composition, and body weight. A secondary objective was to examine the impact of seasonal variability in percentage lipids and lipid composition on PCB concentration in two coexisting Baltic amphipods, M. affinis and Pontoporeia femorata. Polycholorinated biphenyl concentrations tended to be higher in P. femorata than in M. affinis, possibly because of P. femorata's lower respiration rate and larger size. The net accumulation of PCBs was congener dependent and negatively correlated to lipid concentration. The relation between the net concentration change rate of 16 PCB congeners over time and log Kow was not significant during the spring and summer months, a time when lipid accumulation and strong growth occur. In contrast, the net concentration change rate of the corresponding PCB congeners over time during autumn and early winter, that is, from the period before gonad maturation to the period after mating and early embryogenesis, showed a significant relation to tog Kow (r2 = 0.62, p < 0.001, n = 16). During the latter period, amphipod lipid weight was reduced while the PCB body burden increased. Results strongly indicate that elimination rather than accumulation is the main process controlling amphipod body burden. This pattern results in a transfer of PCBs from the female to the developing embryos, which is reflected in high PCB body burden in newly released offspring. PMID- 14552017 TI - Population-level ecological risk assessment of planar polychlorinated aromatic hydrocarbons in great cormorant (Phalacrocorax carbo) around Tokyo Bay, Japan. AB - Assessment of population-level ecological risk posed by planar polychlorinated aromatic hydrocarbons (p-PCAHs; including polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and dioxintike polychlorinated biphenyls) in sediment of Tokyo Bay (Japan) and rivers via fish ingestion to the great cormorant (Phalacrocorax carbo) population was conducted by means of a probabilistic approach. Population decline risk was used as an indicator of population-level effects and compared with other indicators of effects. The increment of egg mortality risk posed by current p-PCAH levels was estimated to be 11.7%. This risk was interpreted in terms of both the increase of the risk of population decline in a 10-year period on a recently abundant cormorant population, and the reduction in population growth rate (r). Population decline risks of 20% and below were estimated to be 16% for the reference population and 32% for the exposed population, whereas the reduction in r was estimated to be 10%. The risk expressed in terms of population viability is a more susceptible measure and a more easily understandable indicator than both egg mortality risk as an individual-level risk and the reduction in r. Translating the effects due to pollutants into the risk on population viability will make ecological risk assessment more conductive to risk management. PMID- 14552018 TI - Identifying pollution hot spots from polychlorinated biphenyl residues in birds of prey. AB - Techniques for determining whether patterns of points are random, clustered, or dispersed are well established; however, when the magnitude of the attribute at each location is also important, the situation is more problematic. The concentration of polychlorinated biphenyls (PCBs) in the livers of Eurasian sparrowhawks Accipiter nisus and common kestrels Falco tinnunculus has been determined for birds from all over Great Britain for several decades and forms a unique database. When mapped, there appears to be clusters of high values in some parts of the country. If these clusters are truly significant, then they may indicate pollution hot spots and possibly help identify undocumented sources of contamination. What constitutes a cluster is open to debate. We know something about the foraging behavior of birds of prey, but we do not know how many pollution sources (hot spots) there are, how long they persist, or over what area they may disperse PCBs. We used a Monte Carlo simulation approach to determine whether the visually prominent clusters of high PCB residues were significant features or merely illusions. The five largest nonoverlapping clusters (defined in terms of the total PCB concentration) were identified at a range of spatial scales. In addition to the total concentration and the number of observations, the weighted centroid of the clusters and which individual birds were involved were also recorded. This enabled us to determine the scale over which the candidate hot spot was stable. Comparing the magnitude of the observed clusters with those from the trial simulations determined the probability of nonrandomness in the original data set (at each spatial scale). Results showed that some clusters do exist but, in the majority of cases, apparent clusters identified by eye could not be considered an actual aggregation of high concentrations following spatial analysis. PMID- 14552019 TI - A basin-specific aquatic food web biomagnification model for estimation of mercury target levels. AB - In the Willamette River Basin (WRB, Oregon, USA), health advisories currently limit consumption of fish that have accumulated methylmercury (MeHg) to levels posing a potential health risk for humans. Under the Clean Water Act, these advisories create the requirement for a total maximum daily load (TMDL) for mercury in the WRB. A TMDL is a calculation of the maximum amount of a pollutant that a body of water can receive and still meet water-quality standards. Because MeHg is known to biomagnify in aquatic food webs, a basin-specific biomagnification factor can be used, given a protective fish tissue criterion, to estimate total mercury concentrations in surface waters required to lower advisory mercury concentrations currently in fish in the WRB. This paper presents an aquatic food web biomagnification model that simulates inorganic mercury (Hg(II)) and MeHg accumulation in fish tissue and estimates WRB-specific biomagnification factors for resident fish species of concern to stakeholders. Probabilistic (two-dimensional Monte Carlo) techniques propagate parameter variability and uncertainty throughout the model, providing decision makers with credible range information and increased flexibility in establishing a specific mercury target level. The model predicts the probability of tissue mercury concentrations in eight fish species within the range of concentrations measured in these species over 20 years of water-quality monitoring. Estimated mean biomagnification factor values range from 1.12 x 10(6) to 7.66 x 10(6) and are within the range of U.S. Environmental Protection Agency national values. Several WRB-specific mercury target levels are generated, which very by their probability of affording human health protection relative to the federal MeHg tissue criterion of 0.30 mg/kg. Establishing a specific numeric target level is, however, a public policy decision, and one that will require further discussions among WRB stakeholders. PMID- 14552020 TI - A matter of ethics. PMID- 14552021 TI - Visit to Guyana inspires how-to guide for dentists in remote regions. PMID- 14552022 TI - Dental visits by pediatric HIV-infected medical patients. AB - The purpose of the study described here was to assess dental-visit frequency in HIV-infected children as reported by caregivers. The sample consisted of 102 HIV infected children aged 3 to 15 years and their caregivers. The child's most recent visit to any dental clinic for treatment or checkup was reported by the caregiver during his or her monthly medical review. The results were as follows: of the HIV-infected children, 50% had been to the dentist in the past 12 months; an additional 12% had been seen 13 to 36 months prior to the inquiry; and 38% had never visited a dentist. Of children 3 to 7 years old, the figures were 39%, 6% and 55%, respectively. For youth ages 8 to 15 years, figures were 60%, 17% and 23%. Within the racial/ethnic groups, 36% of African-Americans, 41% of Hispanics and 50% of white (non-Hispanic) had never been to a dentist. The researchers believe that the study showed that a significant number of HIV-infected children on public insurance programs receiving consistent medical care had not visited any dental provider. PMID- 14552023 TI - Pemphigus: update for the general practitioner. AB - Pemphigus is a dermatologic disease that can affect both the skin and mucous membranes. Pemphigus affects the oral cavity; the most common form of the disease that is observed clinically is pemphigus vulgaris. Oral lesions may precede skin lesions; therefore, it is imperative that clinicians are aware of the clinical signs and symptoms of this disorder. This article will review the etiology, symptomatology, diagnostic tools, and treatments available to diagnose and manage this disease. PMID- 14552025 TI - You can sell your practice and work too. PMID- 14552024 TI - Primary Sjogren's syndrome. AB - Columbia University's Salivary Gland Center (SGC) has examined more than 6,000 patients with a variety of concerns stemming from salivary gland disease and/or salivary secretory dysfunction. Not unexpectedly, the most common patient complaint centers around symptoms associated with dry mouth. Such patients are usually first seen by the dental practitioner. Because Sjogren's syndrome (SS) causes dry mouth, and because it is a relatively common entity--encountered in about three million Americans--and because the dental profession has become aware of its classic manifestation of xerostomia, patients experiencing SS are referred in increasing numbers to the SGC for evaluation. Therefore, the authors wish to call attention to the methodology used in accurately diagnosing SS and to illustrate its signs and symptoms with a case report. PMID- 14552026 TI - Protect your wealth now and later. PMID- 14552027 TI - Thank God, it's a root canal! PMID- 14552028 TI - Operations for lymphedema. PMID- 14552029 TI - On treatment of peripheral lymphedema. PMID- 14552030 TI - Functional impact of lymphangiogenesis on fluid transport after lymph node excision. AB - When a lymph node is excised, lymphangiogenesis occurs to maintain flow in the affected area. However, a complex network of small vessels replaces the node and these newly formed vessels might increase resistance to lymph transport. To test this in sheep, the popliteal lymph node from one hind limb was removed surgically. The contralateral node was left intact. After 4 to 6 weeks (a period that allowed regenerated vessels to restore flow), a prenodal lymphatic vessel in each limb was cannulated with a polyethylene catheter to permit saline infusion into the node or lymphatic regeneration site. Infusion pressures were monitored from t-pieces inserted between the infusion pump and the point of entry of the catheters in the prenodal ducts. We observed that the flow rate versus perfusion pressure relationships were significantly different in the 2 experimental preparations (node intact limbs, n = 13; node excised limbs, n = 10). In the limbs undergoing lymphangiogenesis, much higher infusion pressures were required to generate a given flow rate. Additionally, the regenerated lymphatic network provided a significantly increased resistance to flow. The data suggested that lymphangiogenesis restored fluid continuity to some extent in the area occupied originally by the popliteal lymph node. However, the transport properties exhibited by the newly formed lymphatics were insufficient to restore flow parameters to their original state. PMID- 14552031 TI - Three dimensional architecture of lymphatic vessels in the tongue. AB - We have three-dimensionally reconstructed the lymphatic architecture of the rabbit tongue using a computer graphic three-dimensional reconstruction method together with histochemically stained serial cryo-sections. Three collecting lymphatic vessels lying in an anteroposterior orientation were identified in the tongue body. A superior longitudinal muscle accompanying collecting lymphatic (SLCL) that lies in the border between superior longitudinal muscle and transverse muscle was identified in this study, in addition to collecting lymphatics in the lingual septum (LSCL) and deep lingual artery accompanying collecting lymphatics (DLCL) that we reported previously. The vertical muscle accompanying collecting lymphatics (VCL) and the transverse muscle accompanying collecting lymphatics (TCL) were also identified as collecting lymphatics that joined the above three collecting lymphatics. Specific regional lymph flow was identified for each collecting lymphatic. A transverse right/left cross flow was identified for the LSCL, while the SLCL served the upper 1/3 of the tongue and the DLCL served the lower 2/3 of the tongue. Each collecting lymphatic that accompanied the internal lingual muscles joined to the SLCL, DLCL and LSCL, and were derived from blind-ended lymphatic capillaries that ran in the endomysium. Internal lingual muscle accompanying collecting lymphatics joined each other in the endomysium, and their blind-ended lymphatic capillaries had no branches. PMID- 14552033 TI - Limb volume estimates based on limb elliptical vs. circular cross section models. AB - Limb volumes, as would be estimated by the widely used right circular truncated cone model (right circular frustum), were analytically compared to volume estimates that would be obtained if limbs were represented by an elliptical cross section. A general expression for the ratio of circular to elliptical limb segmental volumes was developed in terms of the ratio of minimum to maximum limb radial dimensions. Analytical results showed that in general the elliptical representation resulted in smaller calculated limb volumes, with the difference increasing as the ratio of minimum to maximum limb dimension became smaller. However, differences in estimated limb volume between circular and elliptic representations were less than 5% if the minimum to maximum limb dimensions at measured circumference sites were greater than 0.64. It is concluded that although limbs deviate from circularity, the added work of determining minimum and maximum dimensions for each circumference measured, as is needed to employ elliptical models, is warranted only for extreme differences in limb radial dimensions or possibly for research purposes. PMID- 14552032 TI - Three-dimensional changes in lymphatic architecture around VX2 tongue cancer- dynamics of growth of cancer. AB - Many questions remain regarding the mechanism of cervical lymph node metastasis via lymphatic vessels. We report here the three-dimensional dynamics of the lymphatic architecture around tumor during growth of implanted VX2 tongue cancer. The tongue and the deep cervical lymph nodes of rabbits were observed at 3, 7 and 10 days after transplantation of VX2 cancer cells (n = 5 in each group). Lymph node metastasis was confirmed histopathologically. Morphological changes of the collecting lymphatic vessels and lymphatic capillaries were observed, and the number and diameter of these lymphatic vessels were measured within 500 microns around the tumor using the combined method of 5'-nucleotidase (5'-Nase) staining and three-dimensional reconstruction imaging. The VX2 cells were uniformly detected in cervical lymph nodes of each rabbit of the 10-day group. The number of lymphatic capillaries and the diameters of collecting lymphatic vessels around the tumor in the 7- and 10-day groups were greater than in the 3-day group. These capillaries arose by sprouting from preexisting lymphatic vessels and showed a tree-like branching pattern. We conclude that the dynamics of the lymphatic architecture around the tumor, especially the increase in number of capillaries on preexisting lymphatic vessels outside the tumor margin, may be associated with lymph node metastasis. PMID- 14552034 TI - Bulging mesothelial cells of the visceral pleura of the rat mimic the network of subpleural lymphatics. AB - The mesothelial surface of the visceral pleura of the Wistar rat was viewed at high resolution by scanning electron microscopy (SEM). The pleural surface showed exquisite linear arrangements made up of bulging mesothelial cells. They were organized in irregular circles that often presented anastomotic junctures. This arrangement of pleural mesothelial cells mimics the organization of subpleural lymphatics of the lung. A low density of microvilli was seen inside the irregular circles, contrasting with the microvilli-rich mesothelial cells seen on or outside these arrangements. These SEM features of the mesothelium may be related with the formation of microdomains for fluid absorption across the visceral pleura into subpleural lymphatics. PMID- 14552035 TI - Revised "management of information" chapter approved for 2004. PMID- 14552036 TI - Revisions approved to telemedicine standards in "medical staff" chapter. PMID- 14552037 TI - New accreditation participation requirement prohibits use of surveyors as consultants. PMID- 14552038 TI - New accreditation participation requirement governs use of a periodic performance review. PMID- 14552039 TI - Staffing effectiveness standards approved for long-term care, assisted living. PMID- 14552040 TI - Automatically easier. Supply chain benefits from point-of-pull. PMID- 14552041 TI - A call for help. Collaboration with community officials is key. AB - The U.S. government's war on terrorism is a battle with an unseen enemy that uses tactics that are just as elusive. Although such conventional terrorists' assault methods as bombs and other explosives are a major concern, the threat of a biological, chemical or nuclear attack is particularly troubling to the government and those who would have to deal with the aftermath of such an attack. PMID- 14552042 TI - Packaging sterility. Maintaining proper storage is crucial. PMID- 14552043 TI - O. R. and oxygen makes three. Communication can prevent fire triangle. PMID- 14552044 TI - Central part of the team. PMID- 14552045 TI - Moving to a distributed authorship model with a "market orientation". PMID- 14552047 TI - Reaching the halfway mark: half of Americans are now health surfers. PMID- 14552046 TI - What doctors want in Web portals. PMID- 14552048 TI - Figuring ROI: how much is your site worth to your organization? AB - How that Web sites are playing more prominent roles in healthcare organizations' revenue-generation strategies--and budgets--the question of return on investment becomes more important, too. Here's how the Emory Healthcare system in Atlanta planned its site upgrade with the need for financial measures in mind. PMID- 14552049 TI - Historical development of endoscopy. PMID- 14552050 TI - Arthroscopy of the temporomandibular joint: technique and operative images. AB - Over the years, little has changed in the preoperative indications and treatment for these patients. They must have failed a course of nonsurgical care, including physical therapy and medication up to 6 months. They must have had imaging done confirming intra-articular joint pathology (disc displacement, disc immobility for the most part, and joint effusions in some instances). Important technical issues include the use of two cannulas placed approximately 1 cm apart. Cannulas are not routinely placed initially into the anterior recess. Heavy downward pressure applied to the posterior ligament with a blunt obturator is used to assure mobility of the disc, and electrocautery or laser energy is used on the surface of the posterior ligament synovium for hemostasis and denervation (pain control). Corticosteroids are used infrequently. To avoid scuffing or damaging the joint surfaces unnecessarily, lysis is done using triangulation methods. Advanced instrumentation is used as the need arises but not used routinely. These patients can expect a predictably successful outcome with minimal recovery time and complications. PMID- 14552051 TI - The endoscopic brow and midface lift. AB - Improvements in technology have increased the level of patient care in all aspects of medicine and surgery. This is no less true in the area of cosmetic surgery. The use of endoscopy has led to improved aesthetics with respect to postoperative scarring, decreased healing time for patients, and an increase in overall patient satisfaction. Because the endoscopic brow and midface lift accomplishes the three primary goals of facial cosmetic surgery (restoration, rejuvenation, and enhancement) while adhering to the concepts of working from inside out and repositioning rather than excising, it could be concluded that there is no longer any indication for either the coronal or trichophillic brow lifts. An argument could be made that a direct brow lift may be indicated in cases of extreme brow ptosis with deep frontal rhytids to allow concealment of the scar; however, even moderate-to-severe brow ptosis can be corrected endoscopically when deep forehead rhytids are not present. PMID- 14552052 TI - Functional endoscopic sinus surgery. PMID- 14552054 TI - Endoscopic orbital surgery. AB - Various applications of endoscopy to orbital surgery are described in this article. The endoscopic approach offers improved surgical exposure, enhanced illumination, simultaneous visualization of operative field by multiple members of surgical team, and superior cosmesis compared with conventional surgical interventions. The procedure can be performed with minimal morbidity. The endoscopic orbital surgery, however, is technique dependent and should be performed only by surgeons who are familiar with orbital anatomy and basic endoscopic technique. It would be beneficial to surgeons to have previous experience in the traditional technique before adopting endoscopic orbital surgery to their armamentarium. PMID- 14552053 TI - Endoscopic mandibular condyle fracture repair. AB - Treatment of mandibular condyle fractures remains a controversial issue. Arguments center on the relative merits of open versus closed treatment. In the past decisions were largely based on philosophy, anecdotal experience, and retrospective case series with short follow-up. Well-designed studies have now begun to appear in the literature and suggest improved results after open, anatomic reduction and fixation. Many surgeons are still hesitant about liberally applying the open approach due to the resultant facial scarring and the risk of facial nerve injury. Developments in endoscopic technology have recently been applied to facial fracture repair. The endoscopic approach to mandibular condyle fracture repair reduces the risk of facial nerve injury, and dramatically reduces facial scarring, compared with standard open approaches. We feel that the reduced morbidity of the endoscopic approach may allow the benefits of anatomic reduction and rigid fixation to be offered to a larger proportion of patients with mandibular condyle fractures. Technical and technological advances are expected to aid in the dispersal of these techniques in the future. PMID- 14552055 TI - Frontiers in maxillofacial endoscopic surgery. AB - The use of endoscopically assisted techniques allows limited incisions for the open reduction of condylar process fractures. Intraoperative control after fracture reduction in areas of limited exposure and visibility is possible. In addition, the risk of facial nerve damage and extensive visible scars can be reduced by using minimally invasive endoscopic techniques. Although the use of endoscopically associated techniques in maxillofacial surgery offers many options and advantages, indications for open treatment of maxillofacial trauma have not changed. The repair of severely dislocated and comminuted fractures of the facial skeleton and major reconstruction of craniomaxillofacial structures still require extended exposure with traditional intraoral and extraoral approaches. PMID- 14552056 TI - [West Nile virus: a new challenge?]. AB - West Nile Virus (WNV), a member of the family Flaviviridae, was first isolated in 1937. Since the original isolation of the WNV outbreaks have occurred with increase in frequency of cases in humans and horses, apparent increase in severe human disease and high avian death rates. In 1999, 2000 and 2002 outbreaks of the WNV encephalitis were reported in horses, birds and humans from New York and Canada. Ornithophilic mosquitoes are the principal vectors of the WNV and birds of several species chiefly migrants appear to be the major introductory or amplifying host. The pattern of outbreaks in the old and new world suggests that viremic migratory birds may also contribute to movement of the virus. If so, Central America, Caribbean Islands and countries of South America including Venezuela, are in potential risk for suffering a severe outbreak for WNV, since several species of birds have populations that pass trough New York and cross the western north Atlantic or Caribbean Sea. It is important the knowledge of the ecology of WNV as well of the efficacy of control efforts in order to minimize the public health impact in these countries, where all population is susceptible to this infection. PMID- 14552057 TI - [Eating behavior disorders. Prevalence and clinical features in adolescents in the city of Maracaibo, Zulia State, Venezuela]. AB - The purpose of this study was to identify the prevalence and clinical characterization related to eating disorders in adolescents from both sexes. In order to do so, the diagnostic criteria and psychological symptoms described by the DSM-IV were used. The sample for this study was constituted by 1,363 adolescents, taken from a population of 23,831 students, who were randomly selected. The sample was stratified taking into account whether they came from a public or a private school and the school levels (middle and high school). The A.B.A checklist was administered to the subjects. This checklist was designed to identify the symptoms that characterize each of the disorders that are the main focus of this study, using DSM-IV criteria. Results from this study show a prevalence of 0% for anorexia nervosa, 1.58% for bulimia nervosa, and .66% for binge eating disorder. Clinical characterization and psychological symptoms related to these disorders show distinguishing features that give distinctive traits to the subjects of this study. PMID- 14552058 TI - [Prenatal maternal blood screening for the detection of fetal chromosomal abnormalities: clinical importance of the rate of false positives]. AB - Maternal serum screening to identify fetal aneuploidies is now routinely offered during the second trimester of pregnancy in developed countries. The purpose of this prospective study was to assess the value of maternal serum screening between 15 and 20 weeks of gestation to detect fetal aneuploidies and to determine the false positive rate (FPR). Blood samples were collected from 1,062 pregnant women between 15 and 20 weeks of gestation. Samples were assayed for alpha-fetoprotein (AFP), free beta human chorionic gonadotropin (beta-hCG) and unconjugated estriol (uE3). Medians were established at each week from 200 normal, singleton pregnancies. Second trimester risk was calculated using the maternal age and different combinations of AFP, beta-hCG and uE3. Screening results calculated by likelihood ratio to be equal to or greater than 1:270 were considered positive. If the gestational age was confirmed by ultrasonography, genetic counselling and amniocentesis were offered. Ten fetal chromosomal abnormalities were detected with maternal serum screening. Sample's size does not allow a correct detection rate estimation, but false positive rate (FPR) was found to be 6.5%. This FPR has a clinical application. At a cut-off of 1:270, second trimester screening best results were obtained using a combination of all three biochemical markers. These results confirm the efficacy of maternal serum screening for fetal chromosomal abnormalities with a low FPR. The measurement of AFP, beta-hCG and uE3 is an effective prenatal screening test. PMID- 14552059 TI - [Frequency of subclinical hypothyroidism among healthy children and those with neurological conditions in the state of Merida, Venezuela]. AB - The purpose of this study was to investigate and to compare the frequency and possible aetiology of subclinical hypothyroidism (SH) in healthy and sick children developing neurological disorder changes. One hundred and eighty-seven male and female children between 1 month and 4 years old, 64 with and 123 without neurological disorders, were studied in the state of Merida, Venezuela. Serum levels of thyrotropin (TSH), free thyroxine (FT4) and urinary iodine were measured by immunofluorescence and by the Sandell-Koltoff's method. Children were diagnosed as having SH if they had high levels of TSH and normal levels of FT4. Antithyroglobulin and antiperoxidase antibodies were measured in children with SH. To establish the frequency of SH, the TSH reference levels on the commercial kit (> 3.8 uU/mL) and the TSH reference levels of our group of healthy children were used. The latter was calculated (X + 2SD) from the values of TSH in the 123 healthy children (> 4.98 uU/mL). The frequency of SH in all children was of 15% when the TSH level from the commercial kit was used and of 6.4% when it was of our group of children (p < 0.001). The frequency of SH value was of 17.2% in children with neurological disorders and of 13.8% in healthy children when the commercial kit's TSH level was used. The frequency of SH in children with neurological disorders was of 7.8% and of 5.7% in healthy children, according to our TSH reference level. This difference was not statistically significant. No significant differences were found in the urinary iodine levels or in the presence of thyroid autoantibodies among the two groups of children with or without SH. There was no association between iodine urinary levels, presence of SH and neurological disorders. We conclude that: 1. In order to make suitable SH level diagnosis in children, we must establish our own levels of reference for TSH. 2. The frequency of SH in children from Merida state is high, being slightly higher in children with neurological disorders. 3. The aetiology of SH is not due to iodine deficiency or immunological factors. PMID- 14552060 TI - [Molecular and serological diagnosis of a dengue outbreak in Coro, Falcon state, Venezuela]. AB - Dengue virus (DV) is responsible for a spectrum of diseases, from a self-limited fever disease (DF, dengue fever) to the more severe forms of hemorrhagic fever/dengue shock syndrome (DHF/DSS). The aim of this study was the serological and molecular confirmation of an outbreak of dengue in Falcon state, Venezuela. A total of 54 sera from patients with clinical diagnosis of DV infection were analyzed by an enzyme immunoassays developed in Venezuela (ELISA -IgM e -IgG) and by PCR. From them, 78% exhibited DV infection (PCR+ y/o IgM+), 48% exhibited viremia by PCR and 57% were positive to IgM. An interesting observation was the high percent (76%) of patients with past or secondary infection (IgG positive), which included all the patients exhibiting clinical symptoms of DHF (n = 8). From the PCR positive sera, serotype 1 was found in 27%, serotype 2 in 54% and serotype 4 in 19%. No serotype 3 was found circulating in this population, although this serotype was already circulating in the nearby island of Aruba. The combination of serological and molecular methods allow us to obtain a fairly precise information of this outbreak. PMID- 14552061 TI - [Subcutaneous microdialysis: a simple technique for monitoring the extracellular biochemical environment. Combination with capillary electrophoresis and laser induced fluorescence detection]. AB - Microdialysis is a simple technique that allows monitoring endogenous or exogenous substances in any extracellular compartment. It has many useful experimental and clinical applications. The sampling of the extracellular fluid of the subcutaneous compartment is especially useful for metabolic evaluation in critically ill patients, pharmacokinetic studies and blood glucose monitoring. We built a subcutaneous microdialysis probe, with a cellulose hollow fiber (13,000 molecular weight cut off, 200 microns outside diameter) glued to stainless steel tubing (26 ga. outside diameter). It was implanted in the subcutaneous tissue of a critically ill child or anesthetized mice to obtain amino acids patterns by means of capillary electrophoresis with laser induced fluorescence detection (CE LIFD). The probe was also implanted in ambulatory volunteers to monitor glucose. The results confirmed that subcutaneous microdialysis is a very simple, inexpensive and not aggressive method with advantages over repeated venipuncture sampling and endovenous microdialysis sampling. The present report shows that subcutaneous microdialysis with the proper analytical technique can be used to monitor the chemical composition of the interstitial compartment in very different preclinical or clinical conditions. PMID- 14552062 TI - [Vertical transmission of Trypanosoma cruzi in Wistar rats during the acute phase of infection]. AB - Research on this form of transmission was carried out on female rats intradermally injected, before mating, with 1 x 10(4) metacyclic trypomastigotes of T. cruzi strains from dog (Pr) and human (YBM). The infected rats, as well as their offspring, were given parasitological, immunological and histopathological examinations during and after gestation. Healthy gestating rats were used as controls. Rats infected with T. cruzi strains showed clear signs of infection between 18 and 45 days post-inoculation (pi). Of 44 offspring from mothers infected with Pr, 4 males (9.1%) showed high parasitemia (432 and 240 tryps./mm3 of blood) at 30 and 40 days after birth, while direct blood examination, hemoculture and xenodiagnosis showed no infection in the other 40, or in the 52 offspring of rats infected with YBM. Anti-T. cruzi antibodies were found in appreciable quantities in infected mothers and in 44 out of 92 (47.8%) of the offspring, with titers that fluctuated between 1:32 and 1:2048 respectively. Histopathological studies of rats sacrificed at the end of gestation showed acute myocarditis and myositis of varying intensity and extent, characterized by abundant inflammatory infiltrate, in some cases associated with nests of amastigotes. The placentas showed moderate cellular infiltrate without parasites in the vascular stroma and amniotic fluid. The offspring of mothers infected with Chagas' disease were reinoculated and showed an acute phase characterized by low parasitemia (p < 0.05); after 60 days, the beginnings of chronic myocarditis and myositis could be observed, of a similar intensity to that observed in offspring born to infected mothers that were subsequently infected. These results confirm that T. cruzi can be transmitted vertically in Wistar rats; that a small number of offspring contract Chagasic infection congenitally; that anti-T. cruzi antibodies can pass from the mother and that these can modify the immune response in the offspring; that the pathogenicity of the strains of T. cruzi plays an important role in congenital transmission independently of origin or geographical location. PMID- 14552063 TI - Human infection by Fasciola hepatica in Venezuela: report of a geriatric case. AB - A new case of human infection due to Fasciola hepatica is reported in the Venezuelan parasitological literature. The patient is an 81-year-old female asymptomatic, with an eosinophilia of 21% and critrosedimentation rate of 26 mm/h and was found during a routine check up. These values were normal at the time of treatment (4 months later), as were several tests of hepatic function, blood chemistry and peripheral blood haematological values. The number of eggs of the parasite were between 90 and 130/g of facees. The patient was successfully treated with the human formulation of trielabendazole at 2 single doses of 10 mg/kg, each separated by 24 h. The same laboratory tests mentioned above did not show modifications, except for a slight increase in the cosinophil counts at 2 and 11 days and erythrosedimentation rate at 11 days post-treatment. Coproparasitological observations carried out at 12, 18, 60 and 120 days post treatment were negative. No clinical symptoms were registered up to 2 months after treatment. The patient most probably acquired the infection at home through the ingestion of commercially available lettuce from an endemic area of bovine fascioliasis, very distant from home, and between 4 and 7 months previous to treatment. We stress the need to investigate F. hepatica infections which he been neglected, probably because of limited knowledge by health workers, lack of specific symptoms and absence of more sensitive diagnostic procedures. PMID- 14552064 TI - Nutritional and physiologic significance of alpha-lactalbumin in infants. AB - alpha-Lactalbumin is the major protein in breast milk (20-25% of total protein) and has been described to have several physiologic functions in the neonatal period. In the mammary gland, it participates in lactose synthesis, thereby creating an osmotic "drag" to facilitate milk production and secretion. alpha Lactalbumin binds divalent cations (Ca, Zn) and may facilitate the absorption of essential minerals, and it provides a well-balanced supply of essential amino acids to the growing infant. During its digestion, peptides appear to be transiently formed that have antibacterial and immunostimulatory properties, thereby possibly aiding in the protection against infection. A novel folding variant ("molten globule state") of multimeric alpha-lactalbumin has recently been discovered that has anti-infective activity and enhances apoptosis, thus possibly affecting mucosal cell turnover and proliferation. Cow milk also contains alpha-lactalbumin, albeit less than human milk (2-5% of total protein in bovine milk), and protein fractions enriched with alpha-lactalbumin may now be added to infant formula to provide some of the benefits of human alpha lactalbumin. PMID- 14552065 TI - Parenteral nutrition-associated cholestasis in neonates: the role of aluminum. AB - Parenteral nutrition (PN) is an essential component in the care of premature and ill infants. The incidence of parenteral nutrition-associated cholestasis (PNAC) ranges from 7.4 to 84%. One substance in PN solutions that has been implicated in PNAC is aluminum. Aluminum loading in animals and humans causes hepatic accumulation and damage. The degree of aluminum contamination of PN solutions has decreased over time, but contamination still significantly exceeds levels that are safe for human neonates. Further study into the relationship between aluminum contamination in neonatal PN solutions and the development of PNAC is necessary. PMID- 14552066 TI - Iron and Parkinson's disease: chelators to the rescue? AB - The essential metal iron has long been implicated in the neuronal damage associated with Parkinson's disease. Recent findings show that iron chelation may prevent the reductions in dopamine and motor disturbances associated with this disease, and suggest the need to examine the role of dietary iron and the use of metal chelators in neurodegenerative disorders. PMID- 14552067 TI - Overweight and obesity status are linked to lower life expectancy. AB - It is well known that overweight and obesity are associated with increased morbidity, which translates into increased risk of mortality. Two recent studies quantify the number of years of life lost owing to high body mass index, but more research is needed to determine whether weight loss would result in the prevention of early death. PMID- 14552068 TI - Understanding racial differences in obesity and metabolic syndrome traits. AB - Epidemiologic data demonstrate a positive association between obesity and certain measures related to the metabolic syndrome that differ among individuals of different ethnic/racial backgrounds. The effect of genetic and environmental factors on these differences is not completely understood; however, insightful information can be obtained by taking into account the degree of ancestral admixture in populations. PMID- 14552069 TI - The refeeding syndrome and hypophosphatemia. AB - The refeeding syndrome is an underappreciated entity characterized by acute electrolyte derangements--notably hypophosphatemia--that occur during nutritional repletion of patients with significant suboptimal caloric intake. Adverse effects of hypophosphatemia include cardiac failure, muscle weakness, immune dysfunction, and death. Hypokalemia and hypomagnesemia commonly complicate refeeding syndrome as well; however, this report briefly reviews the clinical manifestations of refeeding-induced hypophosphatemia. PMID- 14552070 TI - Needed: urologic nursing mentors for new grads. PMID- 14552071 TI - Artificial urinary sphincter, Part I: Overview. AB - Urinary incontinence has a tremendous impact on an individual's quality of life and self-esteem. A number of patients will fail both conservative medical as well as conservative surgical treatments in their pursuit to regain urinary control. The surgical implantation of an artificial urinary sphincter (AUS) is a definitive surgical option to reestablish continence. However, there are many challenges that may arise as a patient progresses through the rigorous preparation, surgical procedure, and recovery process. Understanding the history, various indications, and risks of AUS surgery will aid in counseling patients considering AUS. PMID- 14552072 TI - Artificial urinary sphincter, Part II: Patient teaching and perioperative care. AB - As discussed in Part I, effective management of urinary incontinence is a tremendous quality of life intervention. The surgical implantation of an artificial urinary sphincter (AUS) provides a definitive, although invasive, surgical option to re-establish continence. Understanding the intraoperative surgical procedure, postoperative care, and teaching will aid in preparing patients undergoing AUS implantation for successful outcomes. PMID- 14552073 TI - Artificial urinary sphincter case study. PMID- 14552074 TI - Prostate cancer: perspectives on quality of life and impact of treatment on patients and their partners. AB - Prostate cancer is a major health concern for Western men, but little is known about its consequent impact on sexual function for men and their partners. In this pilot study, the effect of the diagnosis and treatment of prostate cancer on sexual function as it affects men and their partners was investigated. PMID- 14552075 TI - Clinical study of a new urine collection bag. AB - A pilot study of an innovative urine drainage bag that defines a new principle regarding dependent gravity drainage was conducted. Patients and caregivers found improvement in activities of daily living, comfort, convenience, safety, and cost savings as noted by the majority of evaluators. PMID- 14552077 TI - Demystifying the clinical nursing research process: the literature review. AB - The literature review is a critical step in designing and conducting a credible study. It might seem tedious at first, but your own ideas will become better developed and thoughtful as you critique what other researchers have done. It will make your study stronger. Thus, don't skim over this part of the process. A good literature review will make the next steps easier. It will also assure that you are studying an important area, that the outcome has clinical importance to your patients, and that writing and reporting the results will be easier. PMID- 14552076 TI - Urologic challenges in the menopausal woman: a case study. AB - Menopausal women present many challenges to the health care provider. The practitioner must be skilled in treating urinary tract infections (UTIs) and assessing the urogenital tract. Multiple factors should be considered even in the absence of certain disease processes such as diabetes or hypertension. A case study of a postmenopausal, diabetic woman is presented. Assessment and treatment interventions are discussed. PMID- 14552078 TI - Coming attractions! New medications on the horizon for erectile dysfunction. PMID- 14552079 TI - External genital and dermatologic examination. Part I: The female patient. PMID- 14552080 TI - Getting ready for certification: pediatric urology advanced practice. PMID- 14552081 TI - Calcium oxalate kidney stones: another reason to encourage moderate calcium intakes and other dietary changes. AB - Conventional medical thought several decades ago was to restrict the amount of calcium intake in individuals with a history of calcium oxalate stones. In the past decade, several studies have suggested that increasing the intake of calcium may actually reduce the risk of calcium oxalate stone formation. The largest randomized trial of diet and stone recurrence was recently completed. Interestingly, individuals that had normal calcium intakes and lower intakes of protein and salt had a significantly reduced rate of calcium oxalate stone recurrence. This recent trial along with several past epidemiologic studies should be discussed with patients at high risk of stone recurrence. Currently, health professionals have a wealth of information that can be distributed to individuals at high risk of nephrolithiasis, and simple dietary recommendations may be one of the best ways to reduce the risk of calcium oxalate stones. PMID- 14552082 TI - Citrate anticoagulation and adverse events. AB - Several patients with heparin intolerance were dialysed with tri-sodium citrate as anticoagulant without acute clinical problems (good tolerance). After some weeks however problems arose. In all patients an alkalosis developed: the pre dialysis bicarbonate level rose progressively from 27 mmol/l to 40 mmol/l. This could be tempered by lowering the dialysis fluid bicarbonate concentration from 37 mmol/l to 25 mmol/l. A second problem was a progressive rise in pre dialysis sodium level from a mean of 136 mmol/l to 150 mmol/l. Adapting the dialysis fluid sodium concentration from 140 mmol/l towards 132 mmol/l could solve this. The third problem was a progressive rise in serum aluminium level in patients from 3 microg/l to 38 microg/l. After excluding water, concentrate, dialysis fluid, drug intake, etc... as possible sources, we controlled the aluminium level in the glass bottle containing tri-sodium citrate. We noted the very high value of 35,300 microg/l. After replacing the glass bottles with polyvinylchloride bags with a negligible aluminium content, the serum aluminium levels returned back to normal. It is known that citrate chelates the aluminium present in the glass of bottles or vials. PMID- 14552083 TI - Monitoring parameters of dialysis dose. AB - In order to deliver a specific dialysis dose (Kt/V) to all patients, their product Kt (urea clearance K multiplied by dialysis time t) should be individually adjusted according to total body water (V) of each patient. With dialysis time being fixed in most centres for organisational reasons, such individualization can be accomplished by individually set blood flow (QB). For a given t, the value of QB also defines the magnitude of the cumulative blood volume (VB = QB*t), i.e. the volume of blood perfused through the dialyser during the whole dialysis time. VB is displayed by every contemporary dialysis machine but not used. The aim of this work was to derive an easy to use approach to QB individualization based on patient's body weight and dialysis time to obtain a desired Kt/V value which would also be easy to check after dialysis by looking at the obtained VB value. Statistically significant correlation was found between the QB-based Kt/V estimation and Kt/V determined by the other two methods demonstrating practical feasibility of the novel approach. Kt/V values obtained with the QB prescribed according to patient's body weight tended to be better in females and patients with higher body mass index. PMID- 14552084 TI - Nursing terminally sick dialysis patients. AB - The number of terminally sick patients (TSP) in haemodialysis (HD) is rising. The personnel that assist these patients wonder if they are sufficiently qualified to help and care for them and if TSP require treatment that is different to the other chronic dialysis patients. The reason for this study was to investigate if caring for TSP psychologically affected the infirmary personnel who looked after them during HD. PMID- 14552085 TI - Thoughts about biofilm in dialysis water systems. AB - What is a biofilm? Basically, a biofilm is formed when a structure or a surface has a growth on it. The growth consists of a community of micro-organisms, which is active. Growth means that there are metabolic processes taking place. The amount of organisms can be low, and at the beginning there is always just a few, in principle, just one. The surface area covered can also be very small. This will change over time due to the nutritional situation. In this paper the phenomena of surface growth (biofilm) will be reviewed and commented on. PMID- 14552086 TI - European guidelines for vascular access: clinical algorithms on vascular access for haemodialysis. AB - Vascular access remains the Achilles heel of end-stage renal disease patients, receiving haemodialysis. The increase in the number of elderly dialysis patients with additional cardiovascular co-morbidities and diabetes mellitus makes the creation and maintenance of functioning vascular access, more difficult and cumbersome. Therefore, the development of guidelines for vascular access management seems logical. A team of multidisciplinary vascular access experts created a set of algorithms covering the whole spectrum from pre-operative vessel assessment, peri-operative access management and post-operative follow up and surveillance. Additionally, a range of various access complications with their diagnosis and treatment options were included in these algorithms. The new European guidelines for vascular access augment the knowledge of vascular access problems and their management. Additionally, the use of algorithms facilitates the decision-making when treating complications by all physicians dedicated to this field of healthcare. PMID- 14552087 TI - Bed-side blind insertion of peritoneal dialysis catheters. AB - Access to the peritoneal cavity is an essential factor for successful peritoneal dialysis. The technique of catheter insertion can influence technique success and patient satisfaction. As compared to conventional surgical laparotomy, a bedside blind insertion technique under local anaesthesia has logistical advantages for the patient, the hospital and the community. This study compares outcomes of both methods in a single centre. A retrograde analysis of a prospectively collected database on all catheters implanted at the University hospital Ghent between 1/1/1998 and 31/5/2002 was carried out. During this period, catheters were implanted either by conventional laparotomy (CL) or by a bedside blind insertion technique (BI) under local anaesthesia. PMID- 14552088 TI - Validation of physical activity measurement for people on dialysis treatment. AB - Chronic illness frequently contributes to diminished levels of activity, a consequent increased dependence on health related support services and reduced quality of life. Surprisingly few studies have described activity among people with end-stage renal disease (ESRD). The current study examined the utility and validity of the Human Activity Profile (HAP) for describing activity in a sample of Australians with ESRD. A descriptive design supported the primary objective of validating the use of HAP for an Australian renal population. Data was collected from 65 adults with ESRD under the care of one regional and one metropolitan renal unit in Victoria. The HAP measures physical activity across a range of activities of daily living, including a dyspnoea scale. The Sickness Impact Profile (SIP) describes activities associated with daily living in the areas of physical, psychological and independence. The HAP, SIP and a questionnaire summarising demographic and diagnostic details were administered while each person was undergoing dialysis and the results subsequently compared. This study demonstrates that HAP is a valid measure of activity levels for people with ESRD undergoing maintenance haemodialysis. Additionally, scores on SIP and HAP indicate that this group have low levels of daily activity. These findings support the use of HAP in research to understand the activity levels of ESRD patients and the factors associated with decline. PMID- 14552089 TI - Evidenced based nephrology care. AB - This paper highlights the importance of evidenced-based practice (EBP) and its implications to patient and health professionals working within nephrology. The need to ensure that practice is based upon best evidence should be a concern for all health care professionals. Confusion sometimes surrounds the concept of EBP and the potential impact this may have upon patient outcomes. It is the intention of this paper to firstly demystify the process of EBP highlighting the important factors to be considered in moving towards EBP in nephrology care. Secondly, the paper draws upon a number of well-conducted research studies, which have important implications for nephrology care. These selected examples of best evidence will highlight further where ongoing work could be focused. Whilst there is much published within current literature concerning research within nephrology it is essential that practitioners are encouraged to develop the skills to critically search literature, appraise and evaluate its usefulness in developing appropriate and meaningful evidence to enhance practice and patient care. PMID- 14552090 TI - Advanced glycation in uraemic toxicity. AB - The Maillard reaction involves the non enzymatic combination of carbohydrates such as glucose with protein aminogroups to yield schiff bases and Amadori protein adducts evolving into irreversible advanced glycation end products (AGEs). This phenomenon, part of normal ageing, is accelerated in diabetes, as a result of hyperglycaemia, and in renal failure, as a consequence of the accumulation of reactive carbonyl compounds (RCOs). AGEs and RCOs are implicated in uraemic toxicity both at the biochemical and the clinical level (dialysis amyloidosis, atherosclerosis, alterations of peritoneal membrane permeability). Reduction of plasma AGEs and RCOs is an interesting avenue to reduce uraemic toxicity. Therapeutic strategies involve dialysis technique (haemodialysis membranes, daily haemodialysis, ultrapure dialysate, RCO free peritoneal dialysate) as well as drugs inhibiting AGE formation (aminoguanidine and the less toxic angiotensin converting enzyme inhibitors or angiotensin receptor blockers). PMID- 14552091 TI - Depression in dialysis patients. AB - Many psychiatric disorders can be seen in patients with chronic renal failure (CRF). Haemodialysis (HD), which is a renal replacement treatment, causes various psychiatric and psychosocial problems. Patients are dependent on treatment and the illness causes various problems. In addition, strict diet and continuous treatment are other stress factors (1,2). Various studies have been published in different regions and countries about the prevalence of depression and the relation between sociodemographic factors and depression in patients treated by continuous ambulatory peritoneal dialysis (CAPD), which has gradually become common in Turkey. However studies, which reflect the authors' region, have become necessary (2,3). PMID- 14552092 TI - Intravenous administration of epoetin in haemodialysis patients. AB - Recombinant human erythropoietin (epoetin, rHuEPO) can be administered to haemodialysis patients intravenously or subcutaneously. Although the intravenous route is the originally approved and used route of administration, subcutaneous administration has been quite common throughout Europe since the introduction of prefilled syringes. The intravenous route has been shown to be as effective as the subcutaneous route, but patients should have adequate iron stores. In addition, intravenous administration is patient-friendly and results in fewer injections, less pain and bruising, and a minimised risk of immunogenicity. Furthermore, intravenous administration of epoetin is convenient for the nurse and is commonly used in Germany, Belgium and the USA. The purpose of this paper is to illustrate the very easy, practical ways of administering epoetin in prefilled syringes during dialysis at machine level, based on the authors' experiences in Belgium and Germany. PMID- 14552093 TI - Awareness of anaemia in diabetes patients. AB - Although the link between diabetes and anaemia has been firmly established in the renal world, patients with diabetes and healthcare workers in this field are clearly failing to recognise many of the common symptoms of anaemia, a key indicator for renal disease. By forging links and instigating an exchange of information, renal health care workers can work with their colleagues in diabetes to raise awareness of the important benefits arising from the early diagnosis and treatment of the anaemia related to kidney disease. PMID- 14552094 TI - Complications of vascular access: results of a European multi centre study of the EDTNA/ERCA Research Board. AB - The pilot project of the Research Board of EDTNA/ERCA handled the management of vascular accesses (VA) in European dialysis centres. In the first part of the study, centre policies related to VA management were investigated. In the second part of the study, individual patients were followed prospectively during one year. This paper reports on several topics of the second part of the project, investigating complications of the VA related to centre, patient characteristics and dialysis techniques used. Complications most frequently observed were thrombosis, stenosis, infection, bleeding and flow problems. Gradually more infections and flow problems were observed if the centre size and the patients/nurse ratio went up. Complication rate was not significantly influenced by age, gender, renal diagnosis, time on dialysis or medication used by the patient. In contrast, the number of vascular accesses in the past and interventions in the VA before first use resulted in an increased number of complications. Nurses have a key role in the prevention, manipulation and outcome of vascular access related complications. PMID- 14552095 TI - A screening test for vascular access recirculation? PMID- 14552096 TI - Interview with Stephanie McCutcheon, FACHE, chief operating officer, Bon Secours Health System, Inc., Marriottsville, Maryland. Interview by Kyle L Grazier. PMID- 14552097 TI - Managing job transitions: thirteen questions for a successful search. PMID- 14552098 TI - The E in marketing: ethics in the age of misbehavior. PMID- 14552099 TI - Perfect storm: organizational management of patient care under natural disaster conditions. AB - Managing uncertainty is an essential attribute of organizational leadership and effectiveness. Uncertainty threatens optimal decision making by managers and, by extension, reduces the quality of patient care. Variation in the work flows of everyday patient caregiving reflects management's steps to control uncertainty, which include strategies for contending with potential disaster scenarios. Little exists in the literature that reveals how management's strategic response to controlling uncertainty in a real disaster event differs from strategies practiced in disaster simulations, with the goal of protecting patient care. Using organization theory, this article presents the application of uncertainty management to the catastrophic flooding of a major teaching hospital. A detailed description of management's strategies for patient rescue and evacuation is provided. Unique aspects of managing uncertainty stemming from a natural disaster are highlighted. Recommendations on organization responses to disasters that optimize patient care, safety, and continuity are offered to managers. PMID- 14552100 TI - Impact of managed care on healthcare delivery practices: the perception of healthcare administrators and clinical practitioners. AB - Managed care has introduced changes, such as cost effectiveness, access to care, and quality of care, to many components of the U.S. healthcare delivery system. These changes have affected how healthcare administrators and clinical practitioners perceive the impact of managed care on healthcare delivery practices. A survey was initiated to explore whether the perceptions of administrators differed from those of practitioners and to discover which organizational variables could explain the difference. A descriptive, cross sectional survey design was used for the target population of administrators and practitioners in high, moderate, and low managed-care-penetration markets. Two investigator-developed instruments--the Managed Care Perceptions Inventory (MCPI) and the MCPI-Demographic--and an intact centralization of decision-making assessment subscale were used for data collection. Administrators had a statistically significant, more positive perception of the impact of managed care on healthcare delivery than did practitioners. When the distinction between administrator and practitioner was not used as a grouping factor, managed care market penetration, nonprofit status, and years in current employment position were factors that had statistically significant associations with a more positive perception of managed care. Based on these findings, both administrators and practitioners have a role in maintaining awareness regarding their perceptions and should work collaboratively to address issues of concern. Similarly, promoting trust and commitment at the organizational level is important. Recommendations for further research are also provided. PMID- 14552101 TI - Assessing the information management requirements for behavioral health providers. AB - Behavioral health agencies will soon implement automated information-management systems to support their administrative, financial, and clinical care functions. Assessing current information-management capabilities and delineating future needs are prerequisite to recommending a specific information technology solution. Quantifying the discrepancy between current information-management capabilities and future requirements highlights the areas of greatest unmet need for information management. Selecting an information system that addresses the most critical areas of unmet need is a prudent purchase decision. This article describes the results of a process to assess the information-management requirements for agencies that were considering implementation of an integrated behavioral health information-management system. The assessment revealed that these agencies already employed automated systems to manage most financial functions and many administrative functions. Few agencies, however, utilized automated systems to manage clinical care functions. Selection of a behavioral health electronic medical record (EMR) effectively addressed clinical care information-management needs without duplicating existing financial and administrative management functions. Also, the EMR included features that addressed some administrative functions for which a discrepancy between current capabilities and future needs was found. Selecting an EMR instead of an integrated behavioral health information system was associated with a significant reduction in information system acquisition costs. PMID- 14552102 TI - Developing a culturally competent workforce: a diversity program in progress. PMID- 14552103 TI - [Regional anesthesia in heart surgery. Expectation or reality?]. PMID- 14552104 TI - [Postoperative patient-controlled analgesia is more effective with epidural methadone than with intravenous methadone in thoracic surgery]. AB - OBJECTIVE: To compare the efficacy and side effects of epidural and intravenous methadone for postoperative patient-controlled analgesia (PCA) after thoracic surgery. PATIENTS AND METHODS: A randomized, single-blind trial enrolling 30 patients distributed in 2 groups to receive intravenous methadone (ivPCA group) or epidural methadone (epPCA group). Patients in both groups were administered a loading dose of 0.05 mg.kg-1 followed by infusion of 0.5 mg.h-1. The patients could self-dose 0.5 mg with a lock-out interval of 10 minutes and a maximum of 4 doses per hour. Patient characteristics, type and duration of surgery and fentanyl dose were recorded. Pain was assessed on a visual analog scale (VAS). Level of sedation, respiratory rate and occurrence of nausea, vomiting and pruritus were also recorded over the first 24 hours. RESULTS: The 2 groups were comparable. Pain was greater in the ivPCA group than in the epPCA group in the second hour (VAS 3.93 +/- 1.9 and 2.4 +/- 1.65, respectively; P < .05) and the third hour (VAS 3.57 +/- 1.65 and 1.5 +/- 1.16, respectively; P < .05). The total dose of methadone administered was 25.34 +/- 5.65 mg in the ivPCA group and 18.82 +/- 3.52 mg in the epPCA group (P < .002). There were no significant differences in side effects. CONCLUSIONS: The results suggest that epidural methadone has an intrinsic spinal effect regardless of whether or not there is extra-spinal action arising from syste mic absorption. Epidural methadone provides a more adequate analgesic effect in less time and at a lower dose. Both approaches provide good postoperative analgesia with few side effects. PMID- 14552105 TI - [Precision and accuracy of the immediate determination of hemoglobin using HemoCueB Hemoglobin in urgent, surgical, and critical patients]. AB - OBJECTIVES: The prevalence of anemia among emergency, surgery and critically ill patients is high. As a consequence, many of these patients receive transfusions of packed red cells, with hemoglobin (Hb) concentration being one of the most widely applied criteria for prescription. Accordingly, this study was undertaken 1) to ascertain the accuracy and precision of point-of-care Hb measurements obtained with the portable photometric HemoCue B-Hemoglobin analyzer (HBH) in comparison those performed with the reference cell counter Pentra 120 Retic (ABX), and 2) to evaluate the potential clinical utility of the HCB. MATERIALS AND METHODS: Patients from postanaesthesic recovery unit (PRU, n = 37), intensive care unit (ICU, n = 43) were enrolled and capillary and venous blood samples were taken; emergency room patients with bleeding (ER, n = 35) were also enrolled and arterial blood samples were taken. Hb concentrations were measured 3 times for each patient, using both the HBH and the ABX analyzers. RESULTS: No significant differences between mean Hb values obtained with the HBH and the ABX were found; nor were coefficients of variation significantly different. The coefficients of correlation (Pearson'sr) between the 2 devices were > 0.95 for both arterial and venous blood samples, whereas the correlations for capillary blood samples were 0.747 for PRU patients and 0.859 for ICU patients. CONCLUSION: Based on the reliable results obtained for venous and arterial blood samples, Hb concentrations determined with the portable HBH analyzer may be highly useful for point-of-care monitoring of anemia and evaluating the transfusion requirements of ER, PRU, and ICU patients. However, the degree of inaccuracy and variability of Hb measurements in capillary blood samples would discourage us from using it in these patients. PMID- 14552106 TI - [Sciatic nerve block by the lateral route at the level of the popliteal fossa with 0.75% ropivacine: advantages of a more proximal approach]. AB - OBJECTIVES: To assess the efficacy of a sciatic nerve block using a lateral approach 11 cm cephalad to the lateral femoral epicondyle for providing surgical anesthesia and postoperative analgesia in scheduled foot surgery (hallux valgus) after a single injection of 30 mL of 0.75% ropivacaine. METHODS: A block was performed in 30 patients using a point of puncture 11 cm cephalad to the most prominent point of the lateral femoral epicondyle in the groove between the biceps femoris and vastus lateralis muscles. Once the nerve had been located with a stimulator (2 Hz, 0.5 mA), 30 mL of 0.75% ropivacaine was injected. Data recorded were time until block, number of attempts, depth at which the nerve was found, sciatic nerve response obtained, and time until the sensory block was complete. We evaluated quality of anesthesia during surgery, duration of postoperative analgesia, and patient discomfort during performance of the block. RESULTS: Time required to perform the block was 4.3 +/- 1.2 minutes and only one puncture attempt was needed in 27 patients. The nerve was located at 5.5 +/- 0.4 cm, with response located in the common peroneal nerve in 18 patients and in the posterior tibial nerve in 12. The time needed to achieve a full sensory block was 19.3 +/- 5.1 minutes. Twenty-six patients (86%) were very satisfied with the anesthetic quality of the block, 2 were moderately satisfied, and 2 were dissatisfied. Postoperative analgesia lasted 19 +/- 3.4 hours. Four patients reported minimal discomfort during performance of the block. No complications were observed. CONCLUSIONS: The sciatic nerve block from a lateral approach 11 cm cephalad to the lateral femoral epidondyle is an appropriate anesthetic technique for foot surgery. It is safe, effective and easy to perform. Infusion of 30 mL of 0.75% ropivacaine provided adequate anesthesia and long-lasting postoperative analgesia for our patients. PMID- 14552107 TI - [Acute pain in "Don Quixote"]. AB - OBJECTIVE: A descriptive analysis of references to pain symptoms in Don Quixote based on linguistic, literary, clinical, and mannerist features. METHODS AND MATERIAL: Don Quixote was read and an entry was created for each description of an injury or illness (group I/I). A second group of entries was created for each medical commentary (group C). This information was complemented using a computer search for the roots of words referring to pain symptoms, with an asterisk placed after each root to locate all word forms used. Data entries with pain references were analyzed in group I/I by examining patient, location, characteristics, causes, associated disorders, remedies, caregivers, and recovery times and in group C by examining the full text. RESULTS: Ninety-one entries were made (64 in group I/I, 27 in group c). References to pain were found in 37 entries (29 in group I/I, 8 in group C). The most commonly used words came from the roots dol (the Spanish root for dolor, or pain, among other related words) (52) and moli (for forms related to Spanish idiomatic uses of molido, suggesting severe injury) (19). All pains were due to injuries. Forty-two occasions of injury (Don Quixote 14, Sancho 9, others 19) and 51 lesions (ribs 5, back 5, other 15, unspecified 26) were mentioned. Known remedies and caregivers of the period were described, but other invented ones were also present. Associated disorders were insomnia, anxiety, rage, melancholy, functional impotence and other sexual dysfunctions. Recovery times ranged from 0 to 8 days. CONCLUSION: Pain references are subject to literary considerations. The style is simple and the descriptions are credible for the period depicted but do not meet clinical standards. Issues of current interest are treated: pain threshold and sensitivity, social influences, the placebo effect, and affective disorders. PMID- 14552108 TI - [Anesthesia with propofol, remifentanil and cisatracurium in renal transplantation]. AB - A 41-year-old woman with end-stage renal insufficiency in peritoneal dialysis for 3 years received a kidney transplant under anesthesia with remifentanil, propofol, and cisatracurium. She had a history of hypertension and was being treated with enalapril, metoprolol and erythropoietin. After anesthetic induction, blood pressure fell significantly and surgery was performed in a context of hemodynamic stability. The postoperative course was good, with a functional graft and adequate diuresis from the start. Anesthetics with minimal residual effects and as little renal toxicity as possible are ideal for use in kidney transplantation. The drugs used in this case had pharmacokinetic and pharmacodynamic properties that make them particularly appropriate for such patients. PMID- 14552109 TI - [Cesarean section on a full-term parturient with convulsive crises]. AB - A 34-years-old woman in her 35th week of pregnancy experienced epileptic seizures and underwent emergency cesarean delivery of a healthy boy under general anesthesia. The patient had no history of epilepsy and the seizures were later attributed to an intracerebral cavernous angioma. She received treatment with phenytoin and was asymptomatic 3 months later. Although seizures unrelated to preeclampsia or eclampsia in pregnancy are rare, differential diagnosis must determine the etiology of the crisis. PMID- 14552110 TI - [Anesthesia in Sebastian syndrome: a new hereditary macrothrombocytopenia]. AB - The macrothrombocytopenias make up a heterogeneous group of disease involving thrombocytopenia and giant platelets; other clinical or laboratory findings, such as hereditary nephritis, sensorineural hearing loss, leukocyte inclusions, and cataracts, may also be present. The tendency to bleeding is highly variable and is due to decreased expression of the GP1b-V-IX complex on the surface of platelets, leading to altered platelet-vessel wall and platelet-platelet interactions. The 5 autosomal dominant giant-platelet disorders that are associated with macrothrombocytopenia are May-Hegglin anormaly, Epstein, Fechtner, and Sebastian syndromes, and Alport-like syndrome with macrothrombocytopenia. The mutation responsible is in gene 9 (MYH9) coding for the nonmuscle myosin heavy chain IIA that has been identified in the long arm of chromosome 22 (22q12.3-q13.2). The most recently described macrothrombocytopenia is Sebastian syndrome, consisting of thrombocytopenia with giant platelets and leukocyte inclusions. We report the case of a woman with Sebastian syndrome scheduled for abdominoperineal resection for rectal carcinoma. Preoperative studies revealed isolated thrombocytopenia (35,000 platelets/microL) and a mean platelet volumen of 13 fL. Preoperative prophylactic platelet transfusion was carried out with no adverse events. After a postoperative transfusion of packed red cells, needed because of abundant bleeding, clinical course continued to be satisfactory. The anesthetic implications of this syndrome are not well known because few cases have been reported in the literature, and none was found that describes anesthetic management. Nevertheless, thrombocytopenia and the tendency to bleeding present challenges to the anesthesiologist. PMID- 14552111 TI - [Droperidol and cardiac arrhythmias]. PMID- 14552112 TI - [Intradural anesthesia for an emergency Cesarean section in a parturient with morbid obesity]. PMID- 14552114 TI - [Anterior flexion, a possible malposition of the ProSeal laryngeal mask: role of the introducer]. PMID- 14552115 TI - [Opioids and chronic cancer pain. Toward a consensus document. 8th Local Meeting of the Spanish Society for Anesthesiology and Resuscitation, Barcelona, May, 2002]. PMID- 14552117 TI - Women's health. PMID- 14552116 TI - [Fondaparinux and anesthesia]. PMID- 14552118 TI - MAG reaches landmark settlement with Aetna. Agreement to bring transparency and fairness to payment process. PMID- 14552119 TI - Unique to women. PMID- 14552120 TI - Comprehensive health care for women in Georgia. AB - In summary, comprehensive health care for women begins first with a paradigm shift that acknowledges women's health is more than reproductive health. Second, it requires viewing women's health issues from an across the lifespan perspective. This perspective includes addressing women's health beginning with the formative years, through reproductive health and sexuality to the frail elderly. The OWH has launched a statewide effort, with input from the leadership of Georgia, including an eleven-member advisory council attached to the office to develop a comprehensive women's health plan. The OWH will partner with public and private organizations, academic institutions and other government agencies to devise a course of action that addresses the health needs of all women, of all ages, including racial and ethnic backgrounds, and socioeconomic and education levels. This plan will serve as a guide for those organizations whose interest is meeting Healthy People 2010 broad objective of increasing the quality and years of healthy life for women. PMID- 14552121 TI - Reflections of a "lady doctor". PMID- 14552123 TI - Women and heart disease--what is new in 2003. PMID- 14552122 TI - Flannery O'Connor: life with lupus. PMID- 14552124 TI - The ART of infertility. PMID- 14552125 TI - Ain't I a woman. PMID- 14552126 TI - Good Samaritan Health and Wellness Center. PMID- 14552127 TI - Risky business: addressing the malpractice insurance crisis. PMID- 14552128 TI - [Interdisciplinary pain therapy on an intensive care unit: nursing--key to analgesia]. PMID- 14552129 TI - [Migraine--an illness between facts and fiction: no room for prejudice]. PMID- 14552130 TI - [Children in pain on general medicine units: nurses as pain attorneys support and counsel]. PMID- 14552131 TI - [Psychogenic pain: experiences with chronic pain course]. PMID- 14552132 TI - [Health care reform focal point--the Kiel alternative: what we want (interview by Katrin Balzer)]. PMID- 14552133 TI - [Nursing care expenditures and DRG: transparency pays]. PMID- 14552134 TI - [Calculating cost effectiveness in intensive care nursing: INPULS (Intensive Nursing and and Performance System) clarifies expenditures]. PMID- 14552135 TI - [Performance assessment in intensive care: differences become visible)]. PMID- 14552136 TI - [Back problems in nursing care: preventing disk displacement]. PMID- 14552137 TI - [Self-determination until death]. PMID- 14552138 TI - [Nursing in Europe--Netherlands: learning from our neighbors]. PMID- 14552139 TI - [From case to case: a mountain of questions]. PMID- 14552140 TI - [General nursing practice in the hospital for palliative medicine: where death is a part of life]. PMID- 14552141 TI - [Wound care, 8--Nursing care of patients with amputation: when a body part becomes a phantom]. PMID- 14552142 TI - [History of Jewish nursing in Germany in the context of professionalization]. PMID- 14552143 TI - [Digital decision aids--or is the human already a Homo rationalis?]. PMID- 14552144 TI - [Criteral for assessing quantitative disability in expert determination of functional physical disorders. Review of the literature]. AB - The assessment of individuals with somatoform disorders seeking payments or compensation is a major challenge for psychiatrists, insurers and the social welfare system. It is difficult to examine these disorders objectively and to quantify the impairment people experience in their work or private life. In order to develop more rational criteria for the assessment of these patients, we first reviewed the relevant literature and extracted the criteria mentioned by the respective authors. We then grouped these criteria in clinically plausible dimensions in order to develop a list of parameters that could help psychiatric experts to analyse the impairment more objectively and to help institutions in evaluating the assessments. PMID- 14552145 TI - [Surgical accident expert evaluation of dislocation of the cervical vertebrae]. AB - Evaluation of a cervical-vertebra distortion, respectively with prolonged progress and/or degenerative concomitant symptoms, places high demands on the preparatory clerk as well as on the executing adviser. Complete file preparation can best be described as the task of a "hunter-gatherer". The adviser has to determine the actual accident sequela on the basis of delineated and documented complaints with investigative meticulousness. Additionally, non-surgical evidence has to be introduced and treated specifically. PMID- 14552146 TI - [Pitfalls in medical expert evaluation. Of "credible" probands and "convincing" experts]. AB - Periodical reading of medical expert opinions reveals deficient assessment knowledge on the part of German physicians. Six characteristic cases are presented and discussed. The instructive examples show how careless doctors risk their reputation. Inadequate expert opinions are useless. Courts cannot judge correctly, administrations cannot decide in time, insurance companies cannot regulate reasonably. Finally inferior assessments harm more than they serve- especially the doctor's reputation. PMID- 14552147 TI - [Generalized anxiety disorders in primary medical care]. AB - Based on new empirical findings in a large-scale primary care study, the quality of care for the most chronic and debilitating anxiety problem, generalised anxiety disorder, is examined. Following a brief introduction of this disorder, the core findings of the GAD-P study (generalised anxiety and depression in primary care) with more than 20,000 patients of 558 family doctor practices are summarised and measures to improve the quality of care of patients with generalised anxiety disorder, a disorder which is rarely adequately treated, are discussed. This paper particularly emphasises the standard use of time-efficient diagnostic screening instruments, because improved recognition and diagnosis is the prerequisite for appropriate treatment. Further the role of the media to increase awareness of this disorder as well as patient education materials to improve compliance and to enhance treatment outcome effects are highlighted. PMID- 14552149 TI - Sydney or do you prefer to drill? PMID- 14552150 TI - Digital transversal slice imaging in dental-maxillofacial radiology: from pantomography to digital volume tomography. AB - With digital transversal slice imaging (TSI) as a supplementary program to pantomographic devices and digital volume tomography (DVT), images of the viscerocranium are now possible in several planes. The two procedures, TSI and DVT, are complementary. They should be applied where the indication justifies it in dentistry and maxillofacial surgery for diagnostic and forensic purposes. To minimize patient exposure to radiation, computer tomography (CT) should only be used in complex situations of the viscerocranium where soft tissues are involved or in tumor diagnosis. PMID- 14552148 TI - [Risks in mountaineering and trekking]. AB - Mass tourism in the mountains is on the increase. The climbers often do not know much about the possible risks which include death, invalidity and sickness. In this article, risks such as AMS (acute mountaine sickness), HACE (high altitude cerebral edema) and HAPE (high altitude pulmonale edema) are shown. These facts should be taken into account for risk assessment in the private insurance industry. PMID- 14552151 TI - A comparison of two intraoral CCD sensor systems in terms of image quality and interobserver agreement. AB - PURPOSE: Two intraoral CCD sensor systems were compared with respect to image quality and interobserver agreement. MATERIALS AND METHODS: Forty intraoral images from each of the sensors (Trophy RVG5, Sirona Sidexis) were examined by 4 observers in terms of anatomical landmarks and general image quality. The images were divided into 2 groups--filtered and unfiltered. Kappa values for interobserver agreement were calculated for all assessed points. RESULTS: The Trophy sensor showed better image quality in the filtered group, and the Sirona sensor demonstrated good image quality in the unfiltered group. Moderate to substantial interobserver agreement was achieved. The use of filters increased edge artefacts. Sensor specific artefacts in the Trophy system were observed. PMID- 14552152 TI - Functional quality of molar crown occlusal surfaces in the different design modes of Cerec 2. AB - The goal of the present study was to objectify whether the design modes of "Correlation", "Function", and "Extrapolation" of the Cerec 2 system are suitable for generating a functionally perfect occlusal surface of a first mandibular molar crown. For this purpose, an occlusal surface was waxed up according to the biomechanical concept of MH Polz, and this served as the reference occlusal surface. Four different Cerec 2 comparison crowns (produced in the laboratory, Extrapolation mode, Correlation mode, Function mode) were produced and the positions of the cusps were measured. It was shown that the Correlation mode delivers the most accurate results, and the manually produced occlusal surface also showed good results. The occlusal surfaces of the crowns produced according to the Extrapolation and Function mode were clearly inferior. PMID- 14552153 TI - Cognito--management software for functional risk determination. PMID- 14552154 TI - Dentin adhesives create a positive bond to dental hard tissue. AB - The development of adhesive systems is briefly presented. The advantages of adhesively bonded ceramic restorations are discussed, as are the steps involved in creating an adhesive bond to ceramic. PMID- 14552155 TI - Zirconium oxide restorations with the DCS precident system. AB - The DCS Precident system is suitable for the dental practice and enables the processing of different materials and dental structures. The surface is acquired precisely and without contact by means of a semiconductor laser. Easy-to-use software is employed for the three-dimensional design of the restorations. An almost fully automatic milling machine efficiently mills the framework. A clinical case is documented. PMID- 14552156 TI - Esophagitis and esophageal strictures. AB - Esophagitis and esophageal strictures are important causes of esophageal disease in dogs and cats. Clinical suspicion is created when the clinician recognizes the clinical signs suggestive of esophageal disease and accounts for historical information and physical examination findings. Once suspected, the diagnosis of esophagitis and esophageal strictures is a fairly simple one in most cases. Although the benefit of diminishing secretion of gastric acid in patients with esophagitis is unquestioned, other questions regarding adjunctive medical treatments, such as sucralfate and glucocorticoids for dogs and cats with esophagitis, have not been answered through appropriate clinical studies. Esophageal strictures are readily treated with balloon dilation or esophageal bougienage, and clients can expect most patients to become functional, although dietary change may be necessary. PMID- 14552157 TI - Canine gastritis. AB - Gastritis--inflammation of the stomach--is a frequently cited differential yet rarely characterized diagnosis in cases of canine anorexia and vomiting. Although the list of rule-outs for acute or chronic gastritis is extensive, a review of the veterinary literature reveals fewer than 15 articles that have focused on clinical cases of canine gastritis over the last 25 years. The dog frequently appears in the human literature as an experimentally manipulated model for the study of endoscopic techniques or the effect of medications on gastric mucosa. In the veterinary patient, cases of acute gastritis are rarely pursued with the complete diagnostic armamentarium, and cases of chronic gastritis are rarely found to occur as an entity isolated from the rest of the gastrointestinal tract. This article focuses on those findings most clinically relevant to cases of canine gastritis in veterinary medicine. PMID- 14552158 TI - Gastric dilatation-volvulus syndrome in dogs. AB - Gastric dilatation-volvulus is a medical and surgical emergency that principally affects large-breed dogs. Surgical treatment should be undertaken as soon as the patient has been stabilized with fluid therapy and decompression. A gastrectomy might be required if the stomach is becoming necrotic. A gastropexy is required to prevent recurrence. PMID- 14552159 TI - Gastrointestinal motility disorders and gastrointestinal prokinetic therapy. AB - Gastrointestinal motility disorders represent a diagnostic and therapeutic challenge. Disorders of gastrointestinal motility may result in accelerated transit, delayed transit, impaired relaxation, or inappropriate relaxation. The delayed transit disorders are the most important motility disorders of companion animals and may involve the esophagus (hypomotility and megaesophagus), stomach (delayed gastric emptying), small intestine (postoperative ileus and intestinal pseudo-obstruction), or colon (constipation and megacolon). PMID- 14552160 TI - Bacterial-associated diarrhea in the dog: a critical appraisal. AB - The clinical documentation of enteropathogenic bacteria causing diarrhea in dogs is clouded by the presence of many of these organisms existing as normal constituents of the indigenous intestinal flora. The diagnosis of a putative bacterial enteropathogen(s) in dogs should be made based on a combination of parameters, including signalment and predisposing factors, clinical signs, serologic assays for toxins, fecal culture, and PCR. Relying on results of fecal culture alone is problematic, because C perfringens, C difficile, Campylobacter spp, and pathogenic and non-pathogenic E coli are commonly isolated from apparently healthy dogs [10,13,33]. Nevertheless, culture may be useful in procuring isolates for the application of molecular techniques, such as PCR, for detection of specific toxin genes or molecular typing of isolated strains to establish clonality in suspected outbreaks. The oversimplistic attempt to characterize bacterially associated diarrhea by anatomic localization of clinical signs should be discouraged, because most of the previously mentioned bacteria have been associated with small and large intestinal diarrhea. Accurate diagnosis of infections may require diagnostic laboratories to incorporate PCR-based assays using genus- and species-specific primers to facilitate detection of toxin genes and differentiation of species that appear phenotypically and biochemically similar. There has been tremendous interest in the application of microarray technology for the simultaneous detection of thousands of genes or target DNA sequences on one glass slide. This powerful tool could be used for detection of specific pathogenic bacterial strains in fecal specimens obtained from dogs in the future. PMID- 14552161 TI - Protein-losing enteropathies. AB - GI protein loss can result from a heterogeneous group of diseases, including lymphangiectasia, IBD, neoplasia, ulceration, intussusception, and histoplasmosis. PLE should be suspected in any hypoalbuminemic patient with no evidence of exudative protein loss, proteinuria, or HI. A minimum laboratory database for the suspected PLE patient should include a complete blood cell count, a biochemical and electrolyte profile, urinalysis (+/- urine protein:cretinine ratio), and pre- and postprandial bile acid determinations. Fecal alpha 1-PI concentrations may be used to confirm the presence of GI protein loss in cases with concurrent renal or hepatic disease. Because PLE is a syndrome and not a specific disease, the most effective therapy must be directed at the underlying cause. Multiple high-quality endoscopic biopsies are sufficient to diagnose most patients with PLE, although full-thickness biopsies are required in some cases. Patients with PLE are often clinically "fragile," and careful symptomatic therapy must be integrated with dietary and medical management strategies in most cases. PMID- 14552162 TI - Feline gastrointestinal lymphoma. AB - Gastrointestinal lymphoma is a common cause of anorexia and weight loss in older cats, with or without vomiting or diarrhea. Most cats are feline leukemia virus negative and feline immunodeficiency virus-negative. Low-grade gastrointestinal lymphoma may be more common than previously thought, and these cats respond better to chemotherapy agents than cats with high-grade lymphoma. The most significant prognostic indicator is initial response to chemotherapy, with cats that survive the initial induction period generally achieving long-term remission. Thus far, investigations into molecular markers and immunophenotyping have failed to identify useful prognostic indicators. PMID- 14552163 TI - Biopsy of the gastrointestinal tract. AB - Gastrointestinal biopsy is a potentially powerful tool, but it is easy to do it incorrectly. If clinicians are careless in performing or submitting biopsies, or if they blindly believe whatever the histopathology report says, they are abdicating their responsibility to the client and patient. Two comments seem most appropriate. First, the goal of endoscopy is not to be able to place the tip of an endoscope in a particular location; rather, the goal of endoscopy is to be able to access a particular location and then take a diagnostic specimen well enough that surgery can be avoided. Second, attention to detail is worth at least as much if not more than technology. PMID- 14552164 TI - Gastric and intestinal surgery. AB - Gastric surgery is commonly performed to remove foreign bodies and correct gastric dilatation-volvulus and is less commonly performed to treat gastric ulceration or erosion, neoplasia, and benign gastric outflow obstruction. Intestinal surgery, although commonly performed by veterinarians, should never be considered routine. The most common procedures of the small intestinal tract performed in dogs and cats include enterotomy and resection/anastomosis. Surgery of the large intestine is indicated for lesions causing obstruction, perforations, colonic inertia, or chronic inflammation. PMID- 14552165 TI - Laparoscopy. AB - Laparoscopy is a minimally invasive technique for both diagnostic biopsy and surgical procedures. After the basic technique of laparoscopy is mastered and the appropriate indications are learned, these procedures become an easy and rewarding addition to any small animal veterinary practice. PMID- 14552167 TI - Diagnosis of pancreatitis. AB - In summary, pancreatitis is common in dogs and cats, but it seems that most cases remain undiagnosed. Serum amylase and lipase activities are useful as a quick screening test for pancreatitis in the dog only. Serum amylase or lipase activity must be at least three to five times the upper limit of the reference range to suggest a diagnosis of pancreatitis. Furthermore, the diagnosis must be confirmed by other diagnostic modalities, and normal test results do not eliminate the possibility of pancreatitis. Abdominal ultrasound is highly specific for pancreatitis in dogs and cats but is not particularly sensitive, especially in cats. Serum cPLI concentration is highly specific for exocrine pancreatic function and is also highly sensitive for pancreatitis. Similarly, initial data would suggest that serum fPLI is the most sensitive and specific diagnostic test for feline pancreatitis. Until further data are available, however, serum fPLI should be used in conjunction with other diagnostic tests to arrive at a diagnosis of feline pancreatitis. Histopathologic evidence of pancreatitis is conclusive for a diagnosis of pancreatitis. In most cases, however, lesions are localized, and the lack of histopathologic evidence of pancreatitis does not eliminate a diagnosis of pancreatitis. PMID- 14552166 TI - Exocrine pancreatic insufficiency in dogs. AB - Pancreatic acinar atrophy (PAA) is by far the most common cause for the maldigestion signs of canine exocrine pancreatic insufficiency (EPI). The ability to diagnose PAA in the subclinical phase before the development of total acinar atrophy and manifestation of clinical signs has offered new possibilities to study the pathogenesis of the disease. Marked T-lymphocyte infiltration during the progression of acinar atrophy and the genetic susceptibility of the disease have been taken as a primary evidence of the autoimmune nature of the disease. The term autoimmune-mediated atrophic lymphocytic pancreatitis is preferred to describe pathologic findings. A single abnormally, low serum canine trypsin-like immunoreactivity (cTLI) concentration (< 2.5 mg/L), in dogs with typical maldigestion signs has been shown to be highly diagnostic for clinical EPI and is found in dogs with end-stage PAA. Repeatedly subnormal cTLI values (2.5-5.0 micrograms/L) in dogs with no clinical signs of EPI are valuable markers of subclinical EPI and highly suggestive for partial PAA. The primary treatment of EPI is supplementing each meal with pancreatic enzymes. The long-term treatment response for the nonenteric-coated enzyme supplements has been found to be good in half of these dogs, but the response varied considerably. PMID- 14552168 TI - [Influence of topically applied iodides on urinary iodine excretion and thyroid function]. AB - PURPOSE: The present study was undertaken, to evaluate whether ophthalmic drug, containing iodine applied to conjunctival sac might result in a significant increase of urine iodine excretion and thus might be indirectly associated with the risk of iodine-induced hyperthyroidism. The aim of the study was to examine the occurrence of biochemically relevant changes in urine iodine excretion and in thyroid function after topical administration of ophthalmic drug containing iodine. MATERIAL AND METHODS: In the study twenty-two patients without a history of thyroid disease were examined. The Vitreolent (Ciba Vision) was applied topically to conjunctival sac four times a day for one month. The urinary iodine excretion, using "Rapid Urinary Iodine Test" (Merck KgaA) and parameters of thyroid function (TSH, FT3, FT4) were measured before starting and after therapy. RESULTS: The data showed, that was a significant increase in urinary iodine excretion at the end of therapy; as far as thyroid function was concerned, neither changes in level of TSH nor thyroid hormones were observed. PMID- 14552169 TI - [Use of indocyanine green for photodynamic therapy of choroidal melanoma- preliminary report]. AB - PURPOSE: To evaluate the usefulness of indocyanine green (ICG) in the photodynamic therapy (iPDT) of choroidal melanoma. MATERIAL AND METHODS: Thirty eight patients with choroidal melanoma were treated with iPDT. Tumor thickness ranged from 2.6 to 4.1 mm in ultrasonography. Before iPDT Ruthenium-106 plaques were used in 18 cases, in 6 cases with additional transpupillary thermotherapy ("sandwich method"), and Iodine-125 plaques in 3 cases. In 11 eyes the iPDT was the only performed treatment. Therapy was performed with 810 nm diode laser after intravenous injection of 25 mg solution of indocyanine green. Six months after iPDT ophthalmological examination was performed with ultrasonography and ICGA images evaluation. RESULTS: The baseline ICG study showed pathological intrinsic vasculature in all examined cases. Six months after iPDT changes in microcirculation, as well as significant decrease of tumors thickness in ultrasonography (mean 38%), were detected in all cases. Complete regression of intrinsic vessels was demonstrated by ICGA in 26 cases, and partial regression of pathological vascularization was found in 12 patients. CONCLUSIONS: Our preliminary study suggests, that ICG mediated iPDT can be a new and promising way, to treat choroidal melanoma. PMID- 14552170 TI - [Treatment of intraocular metastatic tumors]. AB - PURPOSE: Analysis of the clinical picture of intraocular metastatic tumors and the results of the treatment with various methods. MATERIAL AND METHODS: Between 1994-1997 intraocular metastatic tumors were diagnosed in 14 patients (19 eyes). There were 13 females and 1 male, aged 28 to 69 years (average 50). The primary tumor in 8 patients developed in the breast, in 4 cases in the lungs, 1 in the brain, and 1 in the kidney. In all patients the primary tumor was excised, then chemotherapy was applied in 9 cases, radiotherapy in 3 cases, and hormonal treatment in 2 cases. In 7 patients the metastatic process concerned also other organs: bones, liver, lungs, hypophysis, and lymphatic glands. Metastases developed in 10 months to 11 years since the diagnosis and treatment of the primary tumor. Intraocular tumors were the most often located near the optic disc (8 cases), or near the macula (4 cases). There were usually flat tumors (in 12 cases < 5 mm in thickness), creamy white, sometimes with pigment clamping. In order, to confirm the diagnosis of the tumor, USG was always performed and in 8 cases fluorescein angiography. In all cases topical treatment was applied, which consisted of irradiation with ruthenium (106Ru) in 6 eyes (in 1 case two times), laser coagulation in 3 eyes, and thermotherapy (TTT) with diode laser in 4 eyes, combined treatment (106Ru + TTT) in 2 eyes, 106Ru and 125I brachytherapy in one eye. The dose of radiation for the apex of the tumor was 60-90 Gy (av. 65). The eyeball was enucleated in 3 patients, 4 patients received chemotherapy. 2 patients received hormonal therapy, applied together with the topical treatment. RESULTS: In the majority of cases (14 eyes), a flat scar or the significant decrease of the volume of the tumor was obtained. 8 patients died, two are currently observed, the remaining 4 do not come to the control examination, and there is no information as to their fate. CONCLUSIONS: Good results of the treatment encourage further application of brachy and thermotherapy in the treatment of intraocular metastatic tumors. It allows for the conservative treatment of the eyeball, and also useful visual acuity is retained often. PMID- 14552171 TI - [Treatment of subretinal macular hemorrhages with intravitreous injections of tissue plasminogen activator and SF6 gas]. AB - PURPOSE: Of our retrospective study was the estimation of the efficacy of recombinant tissue plasminogen activator (tPA) and intravitreal injections of SF6 gas in cases of subretinal macular haemorrhages. MATERIAL AND METHODS: In 20 patients aged 23-83 years, the subretinal macular haemorrhages were diagnosed, due to senile macular degeneration (9), hypertension (3), myopic retinochoroidal degeneration (3), other retinal heredodegeneration (2), retinal macroaneurysm (2) and trauma (1). We injected the solution of recombinant tissue plasminogen activator--Actilise (Boehringer Ingelheim) into vitreous cavity of these eyes. After lowering IOP, the intravitreous injections of 0.5 ccm SF6 gas were done, and following 24 hours down head positioning were carried out. RESULTS: The initial visual acuity before treatment was: HM or CF, from less then 1 meter in most of our cases (12 patients). The after treatment visual acuity ranged CF from 2 meters to 0.7. The early treatment conditioned better acuity. Even in cases of delayed treatment, the diminishing of central visual scotoma could be awaited. CONCLUSIONS: The new method of treatment in cases with subretinal macular haemorrhages despite of its symptomatic characters seems to be helpful in restoring the central vision in some cases. PMID- 14552173 TI - [Retrospective evaluation of eyes with high progressive myopia in children and youth ten years after Snyder and Thompson's scleroplasty]. AB - PURPOSE: Of the study was, to assess the safety and efficiency of scleral reinforcement after Snyder and Thompson surgery. MATERIAL AND METHODS: The scleroplasty was performed on 129 eyes of 75 children with progressive myopia from 6 to 10 years of age. The control study group included 40 eyes of 25 children with similar age, mean eyeball axial length and refraction. In the control study group scleroplasty was not performed. The main indication criteria for surgery included: severe myopia more than -6.0 D, and the increase in refraction error more than -1.0 D per year. We evaluated the eyeball axial length in all subjects, before time of surgery and ten years after surgery, using Ultrasound Alcon Imaging System. The visual acuity, tonometry, visual field were evaluated as well. RESULTS: In the study group the mean eyeball axial length measured before surgery was 25.95 mm +/- 0.62 mm. Ten years after surgery the length of the eyeball was 26.97 mm +/- 0.64 mm. The average increase was 1.03 mm +/- 0.29 mm. In the control group, at the time when study group children were operated, the mean eyeball axial length was 25.91 mm +/- 0.48 mm, and ten years later it was 28.06 mm +/- 0.59 mm. The average increase in the eyeball length was 2.13 mm +/- 0.3 mm. There was statistically significant difference between the eyeball axial length progression in the study group and the control group, where surgery was not performed. In the study group no serious complications after surgery were reported. CONCLUSIONS: Scleral reinforcement is an effective and safe surgery, that can stabilize the progression of severe myopia in children. PMID- 14552174 TI - [Corneal radius of curvature among people with myopia]. AB - PURPOSE: To determine the behaviour of the radius of corneal curvature among people with myopia. MATERIAL AND METHODS: A total of 167 people with myopia (117 women and 50 men) aged 12 and 51, were examined. The average of age was 24. Routine ophthalmological examinations as well as keratometry were carried out. The data was analysed using the coefficient of rang Spearman's correlation and the coefficient of linear Pearson's correlation. RESULTS: A positive correlation was found between the length of an average of corneal curvature radius and myopia progress (P < 0.007). Any correlation between the age of the people with low, medium, and high myopia and the corneal curvature radius length was not observed. CONCLUSIONS: (1) In the course of myopia the average of corneal curvature radius is reduced. (2) The length of the average of corneal curvature radius does not depend on the age of people with myopia. PMID- 14552172 TI - [Phacoemulsification combined with trabeculectomy from personal material]. AB - AIM: The aim of the study was to establish the efficiency of phacoemulsification combined with trabeculectomy in patients with cataract and glaucoma. MATERIAL AND METHODS: 102 patients (112 eyes) with cataract and glaucoma, who were operated between 1998 and 2001. The follow up time--from 6 month to 3 years. Intraocular pressure (IOP), drug therapy and visual acuity were analysed. RESULTS: Mean intarocular pressure after phacotrabeculectomy dropped from 21.6 mmHg to 14.9 mmHg. The average number of medications dropped from 2.6 to 1.2. Visual acuity improvement was achieved in 93.7% of the eyes. CONCLUSIONS: Phacoemulsification combined with trabeculectomy is safe and efficacious method of treatment of patients with coexisting cataract and glaucoma; shortens visual rehabilitation period and secures long-term postoperative IOP. PMID- 14552175 TI - [Epithelial ingrowth after LASIK--personal experience]. AB - The current incidence of epithelial ingrowth after LASIK varies from 1% to 12%. After second procedure, it increases up to 32%. Known risk factors for this complication include epithelial defects at the time of surgery, or a history of recurrent corneal erosions, corneal basement membrane epithelial dystrophy, history of ingrowth in the other eye, hyperopic LASIK correction, flap instability, repeated LASIK surgeries. We performed 200 LASIK procedures. In 12 patients we found epithelial ingrowth. The follow up period was 2 years. We applied treatment: lifting and manual removal in 7 cases, phototherapeutic keratectomy in 3 cases. The rest of cases were under observation. Epithelial ingrowth is a relatively rare complication, following LASIK. PMID- 14552176 TI - [Analysis of 24 viral keratitis cases treated by corneal graft from 1998-2000]. AB - PURPOSE: To determine the clinical character of patients with keratoplasty after viral keratitis and comparison of pre- and postoperative visual acuity. MATERIAL AND METHODS: The study involved 24 patients treated using corneal graft after viral keratitis complications. Sex, age, permanent residence, duration of disease, previous pharmacological treatment, indications to corneal graft and postoperative functional results were evaluated. RESULTS: In 24 cases 16 (66.7%) were males in age 38-75 years old. Eight (33.3%) of the 24 cases were females in age 12-78 years old. Visual acuity improvement after corneal graft was observed in 23 patients. There was no visual acuity improvement in one case with considerable degree of optic nerve lesion. CONCLUSIONS: The results of these examinations showed, that keratoplasty after viral keratitis complications were more frequent in males, especially in 40-59 age range, professionally engaged. Statistical analysis of these cases showed good prognosis of visual acuity improvement after surgical treatment. PMID- 14552177 TI - [Changes in lens and retina and parameters of lipid metabolism and glycosylated hemoglobin levels in young patients with diabetes]. AB - PURPOSE: The aim of the work was, to find out the relations between the lipid profile and glycosylated hemoglobin in blood and eye changes in the diabetic young people. MATERIAL AND METHODS: There were 39 patients (19 women and 20 men), aged 14-27 years (mean 19.5), treated for diabetes during the period 6-23 years (mean 9.6). The ophthalmological examinations have included anterior segment and lens, as well as the fundus of the eye. There also have been made the blood serum examinations of cholesterol (CHC), LDL-CHC, HDL-CHC, trigliceride (TG), apolipoproteins B (ApoB) and glycosylated hemoglobin (HbA1c). RESULTS: Basing on these examinations it has been stated that: 1. the lens changes were observed in the patients of increased LDL-CHC level and ApoB, who suffered from diabetes shorter than 10 years, while in the patients suffering longer, lens changes correlate with the high level of HbA1c; 2. the eye fundus changes correlated substantially in the patients with the high levels of HbA1c, TG, HDL-CH (negatively), LDL-CH and ApoB, suffering less than 10 years; in the patients suffering longer, correlation was also with the increased level of HbA1c, CHC and LDL-CHC. 3. The increased levels of lipids were noted in the patients with the high level of HbA1c. CONCLUSIONS: The stated data have indicated, that the eye changes are caused by disturbances of lipid parameters, but besides of the glycaemia disorders. The diabetic duration does not matter for the lipid profile and HbA1c level, but it is important for the occurrence of eye fundus changes. The HbA1c essentially influences on the occurrence of eye fundus changes, but in the longer suffering patients, also on the lens changes. The high HbA1c level shifts the lipid profile towards atherogenic factors. It is purposeful, to decrease its level below 7%, which is accepted as good diabetes control. PMID- 14552178 TI - [Betaxolol for prevention of steroid induced intraocular pressure elevations in patients after radial keratotomy]. AB - PURPOSE: To investigate the effect of 0.25% betaxolol (Betoptic S) in the normal myopia eye, on steroid-induced intraocular pressure (IOP) elevations in patients undergoing radial keratotomy. MATERIAL AND METHODS: Radial keratotomy was performed on 27 patients (43 eyes). All these patients during the period of 3 months after surgery were treated with 0.1% dexamethasone drops in decreasing doses and 0.25% betaxolol (Betoptic S) drops twice a day. Ophthalmic examination and IOP measurements were made on 1, 2, 3, 14, 30, 60 day after operation. RESULTS: During 3 months of postoperative period 27.9% of eyes had IOP exceeding 25 mm Hg, or had an increase of IOP of 10 mm Hg or more. CONCLUSIONS: The results of the current study and the study published by the author previously lead to conclusion, that 0.25% betaxolol (Betoptic S) in the normal myopia eye, decreases the risk of steroid-induced IOP elevation in patients after radial keratotomy, but does not prevent it. PMID- 14552179 TI - [Clinical evaluation of provitamin B5 drops and gel for postoperative treatment of corneal and conjuctival injuries]. AB - PURPOSE: Vitamins B group are important ingredients of coenzymes. Provitamin B5 dexpanthenol is necessary in the processes of reconstruction of epithelium, has regenerative and anti-inflammatory properties. Objective of the study was to determine how the wounds of cornea and/or conjunctiva heal upon the application of D-panthenol/the medicine has been developed in the Faculty of Pharmacy of the Medical University of Gdansk/. MATERIAL AND METHODS: The activity of the medicine was tested in 40 eyes in the test group, a 40 eyes in the control group did not receive D-panthenol. The degree of the objective and subjective symptoms was assessed. RESULTS: The differences between the two groups commenced on the second day following the operation. Better effects were observed in patients receiving D panthenol. Congestion and oedema of conjunctiva withdrew, the edges of wounds demonstrated smoothness and better adherence. Subjective feelings improved. CONCLUSIONS: 1. Provitamin B5 contained in 5% drops and 5% gel of D-panthenol effectively accelerates the processes of healing the wounds of conjunctiva and cornea. PMID- 14552181 TI - [Surgery for involutional ectropion--case report]. AB - In this paper authors described methods of surgery in involutional ectropion, surgical anatomy and changes in the lower eyelid due to aging. The surgical treatment of advanced involutional ectropion and its results is presented. PMID- 14552180 TI - [Gel with provitamin B5 applied during tests with the Goldmann triple-mirror]. AB - PURPOSE: The objective of the study was, to determine whether the new medicine developed in the Faculty of Pharmacy of the Medical University of Gdansk in the form of gel containing 5% of provitamin B5, was a good agent for applying into the contact surface of the Goldmann triple-mirror, while examining the retina. MATERIAL AND METHODS: 42 people were subject in the observation. 84 eyes, which had not been operated yet, were tested. One eye was examined by using the gel with provitamin B5/proper name D--panthenol/, the other eye by applying 3% methylcellulose. Congestion of conjunctiva, oedema and defects of cornea epithelium, and substance transparency as well as surface adherence were indicated. RESULTS: One obtained the statistical significance proving the lesser congestion effect, while applying D--panthenol. Better adherence of the preparation to the cornea surface and to the triple-mirror was noticed, which considerably improved the procedure of the very examination. D--panthenol protected and regenerated cornea epithelium. CONCLUSION: The gel containing provitamin B5 is a safe and convenient agent for application during the examination with a triple-mirror. PMID- 14552182 TI - [Non-arteritic anterior ischemic optic neuropathy--case reports]. AB - Nonarteritic anterior ischaemic optic neuropathy usually caused by hypertension, diabetes and hypercholesterolaemia is quite a common disease. Within its symptoms two are the most important: deep and often not reversible visual aquity loss and visual field defects. The decrease of visual aquity may be the first sign of systemic illness, which is noticed by a patient. In many cases the correct diagnosis and anti-oedematous, anti-inflammatory and vascular treatment may improve their visual function and general condition. PMID- 14552183 TI - [Treatment results of treatment for congenital, unilateral microcoria and pupil displacement--case report]. AB - The purpose of this report was to present rarely occurring developmental anomaly of congenital unilateral microcoria and pupil displacement. We present a case of a boy, who underwent a plastic surgery of pupil at age of 3 year. Despite of relatively tardy onset of treatment and existing profound amblyopia (light perception with projection), a full vision recovery was achieved after 4 years of occlusion therapy. PMID- 14552184 TI - [Side effects during treatment with Solu-Medrol for optic neuritis in the course of MS]. AB - The aim of this research is, to show the action mechanism of metyloprednizolon and the side--effects of the Solu-Medrol application in the treatment of demyelinization neuritis of ophtalmic nerve in sclerosis multiplex. In research, there were described the side-effects after second Solu-Medrol bolus dose given to a 44-year-old patient. She has suffered from SM and demyelinization neuritis of ophtalmic nerve with rapid deterioration of visual acuity for 14 years. In the years 1995-2002 there has not been any case of side-effects similar to this patient's, among all the 30 other patients treated with Solu-Medrol. It confirms the usefulness of bolus dose metyloprednizolon therapy applied to patients suffering from ophtalmic nerve demyelinization with rapid deterioration of visual acuity. PMID- 14552185 TI - [Foreign bodies in the orbit misdiagnosed as fracture of the orbital lateral wall]. AB - 54 years old male presented with an injury to the face. As a result of this incident, some pieces of glass were penetrating into the left orbit. Total ophthalmoplegia and hematoma with exophthalmus in the left orbit were observed. The difficulties of radiology and clinical diagnosis was described. The circumstances of accident were wrong reported by the patient. Radiograms and computed tomography (CT) were misdiagnosed. As a result of theses, it was necessary to perform two times operation. The best useful but not ideally in diagnostic was NMR. Consultation and collaboration with oculist, radiologist, neurologist and maxillofacial surgeon is often useful. PMID- 14552186 TI - [Use of intra-stromal corneal ring segments (INTACS) in refractive surgery]. AB - Implantation of intrastromal corneal rings segments is a new refractive procedure. INTACS consists of 2 separate segments placed in the midperipheral cornea, in specially preparing lamellar channels. Polymethylo methacrylate rings, which flatter the anterior curvature of the cornea. The peripheral localization of these rings leaves the central cornea untouched. The 2 main advantages of the INTACS are, that it preserves the corneal center and is removable. Segments serve for the reduction of myopia from -1.00 D to -5.00 D. The refractive correction is modulated by INTACS thickness, which ranged from 0.25 to 0.45 mm. PMID- 14552187 TI - [Formation of congenital defects during ocular development. I. Embryogenesis of the eye and developmental birth defects]. AB - Formation of developmental congenital defects of organ of vision finds its direct basis in the course of eye embryogenesis. Author of this work presents on the base of the literature, the crucial stages of embryonal development of eyeball and its individual structures. At every stage of development, the critical moments are shown, when the disturbance of the developmental process may lead to the origination of different groups of congenital eye defects. The knowledge of the potential period of particular congenital eye defects formation, facilitates the identification of eventual teratogenic factors, that have acted during pregnancy. It enables, to state the activity of gradually discovered genes, that regulate the course of embryonal development of the eye. PMID- 14552188 TI - [Formation of congenital defects during ocular development. II. Genetic background of oculogenesis and developmental birth defects]. AB - The course of oculogenesis is genetically controlled by genes, that are selectively expressed in the developing eye. Many of them, first recognized in studies on model animals, such as fruit fly (Drosophila melanogaster) proved, that they play similar roles in humans. The crucial role in cascade of genetic control of oculogenesis plays a system of genes PAX6-EYA1/SIX3-DACH1. In this work there are described also other genes, that form a network of interactions and regulations, such as: BMP4, BMP7, OPTX2, SOX1, PAX2, PITX2, PAX3, CHX10 and other genes. Describing function of each gene, author presented simultaneously all known congenital defects and syndromes, that are caused by mutations of above mentioned genes, such as: aniridia, anophthalmia, Peters anomaly and others. PMID- 14552189 TI - [Current concepts in the pathogenesis of glaucomatous optic neuropathy]. AB - On the basis of review of available literature, the concept of vascular dysregulation, excitotoxicity in the pathogenesis of glaucoma, and influence of intraocular pressure and immunology system on mechanism of glaucomatous optic neuropathy was described. PMID- 14552190 TI - [Optic nerve neuropathy in the course of giant cell arteritis]. AB - Anterior ischemic optic neuropathy (AION) is the result of infarct of the optic nerve head, caused by occlusion of one or more short posterior ciliary arteries. On the base of different treatment and prognosis there are two forms of AION: arteritic and non-arteritic (NAION). Arteritic ischemic optic neuropathy is caused by giant cell arteritis (GCA). The most typical symptoms are: the sudden and deep vision loss and headache, scalp tenderness, jaw claudication, muscle ache, fever and weight loss. The ophthalmologist usually finds an abnormal pupil, a swollen optic nerve (disc edema), and peripheral or central vision loss (or both). About 70% of cases are not progressive, i.e., the vision remains stable, but reduced. The ESR is usually markedly elevated. Temporal artery biopsy is useful in confirming the diagnosis of arteritic AION. Treatment involves the immediate administration of systemic steroids. Though steroid therapy rarely results in the return of vision, it is beneficial in protecting the fellow eye from vision loss and improving long-term systemic health. PMID- 14552191 TI - [Thrombophilia as a risk factor for central retinal vein occlusion]. AB - Central retinal vein occlusion is associated with systemic conditions, such as arterial hypertension, diabetes, hypercholesterolemia, but also with certain hypercoagulability states (thrombophilia). Growing evidence indicates, that the presence of antiphospholipid antibodies and elevated plasma homocysteine levels predispose to central retinal vein occlusion. In younger patients, factor V Leiden (Arg506Gln) could increase the risk of this disease. Other probable causes of thrombophilia related to retinal vein occlusion include increased levels of plasminogen activator inhibitor 1 and lipoprotein (a), which impair fibrinolysis. PMID- 14552192 TI - [The role of ultrasound examinations in diagnosis of massive suprachoroidal hemorrhage occurring as a postoperative complication of intraocular surgery ]. AB - This article presents importance of the ultrasound examinations (standard A- and B-scan ultrasonography, Colour Doppler and Power Doppler ultrasonography) in early and late diagnosis of massive intraoperative suprachoroidal hemorrhage. Standardized echography can help establish an accurate diagnosis. Echography can determine the location and extent of expulsive hemorrhage, as well as determine the status of the retina and vitreous. Furthermore, differentiation between hemorrhagic choroidal detachment and serous choroidal effusion can be made only by A- and B-scans. Colour and Power Doppler imagings of orbital vessels are relatively new sonography methods of ocular diagnosis. Using them, we can evaluate all blood flow disturbance, which occur in eye with massive suprachoroidal hemorrhage. These methods are also useful in differentiation of retina from other tissues, especially in the presence of opaque media. PMID- 14552193 TI - [Complications after laser refractive surgery]. AB - The laser refractive surgery is very effective method of refractive errors correction. The method is very popular, but serious complications may occur and the visual acuity may decrease. In this article authors show the possible complications after the laser refractive procedures. PMID- 14552194 TI - [Historical outline of deveopments in ophthalmic anesthesiology]. AB - The history of ophthalmic anesthesiology is closely related to the development of ophthalmology. Specific needs during ophthalmic procedures bring about on further improvement of effectiveness of local and general anesthesia, since both methods are in use for ophthalmic surgery. It is hard to decide, which one is the better and more suitable for the patient, as well as the surgeon. Individual choice adequate to the certain patient and certain clinical image requires close cooperation between ophthalmic surgeon and anesthesiologist. PMID- 14552195 TI - [Ophthalmology in the ancient river cultures. Was cataract surgery performed during the time of King Hammurabi?]. AB - The article presents the state of ophthalmological knowledge in the Ancient River Cultures. Medical and ophthalmological information included in King Hammurabi Code is discussed. The author presents a critical analysis of the views related to the hypothesis of possible performance of cataract operation in Babylon during the Hammurabi period. Terminological and medical matters related to this issue are discussed. The original opinion about surgical treatment of symblepharon or pannus trachomatosus is presented. PMID- 14552196 TI - [Genotoxic and ecotoxic effects of urban waste water disinfected with sodium hypochlorite or peracetic acid]. AB - Genotoxic and ecotoxicologic effects of urban wastewater disinfected with sodium hypochlorite or peracetic acid were analyzed. The formation of genotoxic species was studied by determining clastogenic and mutagenic activity of aqueous samples and their extracts with in vivo and in vitro tests, respectively. In particular, we have applied citogenetic tests to Allium cepa roots and Tradescantia inflorescence (Allium cepa test and Tradescantia/micronuclei test) and reversion test to Salmonella typhimurium according to the microsuspension procedure (Kado test). The latter is the method of choice for the analysis of complex matrices due to its high sensitivity and specificity. The mutagenic activity of disinfected effluents was similar to the corresponding untreated wastewater both sampled in four different periods. Therefore, the disinfection process did not seem to contribute to aquatic mutagenicity in the examined range of biocide concentration. The potential toxicity of disinfected wastewater for aquatic organisms was evaluated using Daphnia magna. The acute toxicity of peracetic acid in sewage was 0.4 mg/L (24 h E(L)C50). By comparing this value with peracetic acid concentrations detected in effluents from a pilot plant it is expected that treated wastewater would show acute toxic effects on aquatic organisms. Dissociation compounds (hydrogen peroxide and acetic acid) and possible by products of peracetic acid did not seem to contribute significantly to the toxicity of sewage treated with peracetic acid. PMID- 14552198 TI - [Antiasthmatic drug consumption as an indicator of the prevalence of respiratory pathology in a pediatric population]. AB - The study of disease prevalence requires specific and sensitive indicators, which are hardly gathered at population level. The use of anti-asthma drugs, which are relatively specific for diseases characterised by bronchospasms, has been already experimented in the USA with algorithms describing patients affected by asthmatic disease by their use of drugs. We retrospectively analyse spatial variations of drug use as dispensed by the SSN (National Health Service) to Lombardy children less than 15 years old, between 1st January 1995 and 31st December 1997, estimating prevalence of use as a proxy of disease prevalence. Two algorithms already experimented and a new one have been employed to select children treated with antiasthmatic therapy (R03 of ATC classification) dispensed. Local communities' data were used for basic demography. Number of daily defined doses (DDD) dispensed has been used as indicator of drug quantity. Euro/year per person has been used as indicator of costs. Small area distribution analysis has been performed with a Geographic Information System at local Communities level. Frequency of patients (of a total of 1,252,958 children): with at least one dispensed drug in the three years was 26.6% (SD 10.5), DDD per person per month were 1.21 (SD 0.78), Euro/Year per person 6.84 (SD 3.61); with the association of a anti-inflammatory and a b2-agonist was 8.2% (SD 5.6), DDD per person per month were 3.01 (SD 3.07), Euro/Year per person 15.87 (SD 15.07); with at least 90 days of DDD in the three years was 2.6% (SD 1.3) DDD per person per month were 7.79 (SD 4.48), Euro/Year per person 37.66 (SD 21.83). It must be taken into account that, in the best of cases, these approximation refers to the prevalence of all diseases, characterised by bronchospasm, treated with the selected drugs. Estimated data of prevalence are comparable with those of other authors. Prevalence of drug use appears to have relevant geographical differences. Taking into consideration these comments, prevalence of drug use and consequently the estimated disease prevalence appears to be worryingly high, having as well wide geographical differences. PMID- 14552197 TI - [Hygiene education in the Laureate Course of Dentistry and Dental Prosthetics: evaluation of student knowledge about the risk of infection in a dental practice environment]. AB - Hygiene education for dental students is crucial in order to control and reduce the risk of infections in their future professional practices. The aim of this study was to evaluate the improvement in university students' knowledge of infectious disease prevention, dental instruments and environmental decontamination following the Course in Dental Hygiene and Prevention. 595 questionnaires were submitted to dental students at twelve Italian Universities; 335 were filled out before the Course began and 260 at the end of the same course. The students in both the PRE-course and at the POST-course evaluations demonstrated adequate knowledge regarding the transmission of infections in dental practices, knowledge improved following the Hygiene course. PMID- 14552199 TI - [The Epidemiologic Observatory and the Epidemiology Service: the Campania Region experience]. AB - As an effect of recent Laws, epidemiology will be more and more involved in the assessment of goals in Public Health and policy making, so that the debate about structure and placing of epidemiological function is more than ever actual. In this view, the route of epidemiology in Campania Region is explained, with a detailed description of the history and laws as well as of the structure and functions of both central and local units in charge of epidemiological function; weaknesses and strengths of the current system are fully examined and an hypothesis for an organizational model in the next future is formulated. PMID- 14552200 TI - [Population study of knowledge and practice in the Albanian population regarding water usage and diarrhea prevention]. AB - The worsening of the social-environmental conditions during the period of transition of Albania and an epidemic of cholera (1994) were the cue for the execution of the study. The main objective is the description of knowledge and attitudes of the Albanian mothers about the risk factors for the gastroenteritis. This is a descriptive study of a representative sample (2100 families) of the population who reside in the central area of Albania, in 1996. The sample has been selected by cluster-sampling method. Interviewers trained previously, collected the data with a questionnaire ad hoc. Data elaboration has been carried out with the software Epilnfo 5. 99.7% (2094) of the enlisted mothers, mean age 28.7 years, were interviewed. 2/3 of them reside in rural areas. We considered social-demographic variables and their relation with the generic hygienic norms and those relative to water, foods, animals. The basic knowledge was very insufficient, particularly in rural areas, and in relation with lower social economic index and instruction. The study reflects the state of transition train of the political and social-sanitary situation of Albania in the last decade. Still the knowledge and the attitudes about the alimentary hygiene are inadequate. It seems opportune to face this emergency with campaigns aimed at health education and public health interventions. PMID- 14552201 TI - [Development and validation of a questionnaire measuring hospital consumer satisfaction ]. AB - This study describes the process of development of a new instrument for measuring patient satisfaction with hospital care in Italy. The self-administered questionnaire included a total of 48 items, -contributing to the construction of 8 scales, each of them describing a specific area of interest such as medical care and nursing, several aspects of organisation, comfort and overall outcome of care. Psychometric characteristics of the questionnaire were in terms of acceptability, validity, reliability and the ability to discriminate different levels of satisfaction in various patient groups. Psychometric analyses resulted in a convincing construct validity and reliability, as described by the Cronbach Alpha coefficient estimates [range 0.73-0.95]. The high compliance obtained (67.3%) can be considered a sign of good acceptability of the questionnaire and of its mode of administration. These analyses demonstrate a good performance of the questionnaire in terms of both validity and reliability, suggesting that this instrument can represent a solid base for future applications. PMID- 14552202 TI - [Evaluation of professional exposure to antiblastic chemotherapeutic agents in a Tuscan hospital environment]. AB - The aim of this study, carried out from 1997 to 2000, was to evaluate the exposure to antiblastic drugs of hospital staff (38 nurses of 7 oncological Day Hospital care in Tuscany). To evaluate the internal exposure was used biological monitoring, in particular was determined the concentration of urinary cyclophosfamide (CF); pad was made to evaluate the skin contamination. The contamination of working surfaces was determinate, by wipe-test, to verify the decontamination procedures; were used, as markers, CF and 5-fluorouracil (5-FU). We assessed the permeability thorough gloves usually used by the staff and the degradative activity to these drugs by agents used to decontaminate the working surfaces. The results shows, in urine, value lower than the detection limits, although was found a diffused contamination of the working environment. The results confirmed that NaClO is the best product to decontaminate working surfaces and nitrile gloves the ones with less permeability. PMID- 14552203 TI - [Quality, success, and appropriateness of using home care through the Local Health Station No. 13 in Novara]. AB - The research is aimed at evaluating the quality, appropriateness and outcomes of home-care services in the District of Novara of ASL 13, by experimenting a multidimensional model of analysis based on the current clinical records. In this study, we analysed 102 cases of admittance to Integrated Home-care Services (IHS) (50 women and 52 men; average age 72.5). The analysis of the levels of activity highlighted a widespread under-use of the service and an insufficient quality of the clinical records. The management of patients was appropriate in 43.14% of the cases, in terms of intensity and complexity of the received care. 50% of the subjects did not present such characteristics as to need IHS, while 7.86% of the patients evidenced dubious appropriateness, according to an adequate healthcare intensity unaccompanied by a sufficient complexity of the interventions. As far as outcomes are concerned, we were able to highlight how relatively few subjects actually reached the objective of the healthcare plan (26.47%). On the whole, the experimented model of analysis allowed us to trace an effective picture of the quality of the service, by highlighting its critical elements and helping in finding specific actions of improvement. PMID- 14552204 TI - [Informed consent forms in medical practice: the results of a knowledge survey]. AB - "Federico II" University of Naples Hospital Management's Staff considered 47 informed consent forms in order to evaluate quality and quantity of the information transmitted to patients. Every form, distinguished by procedure (surgical, pharmacological, anesthesiologic or related to blood transfusions) was evaluated for the accuracy and validity of information related to: diagnosis, prognosis, diagnostic and therapeutic opportunities, expected benefits, adverse side effects, iatrogenic risks, privacy policy on personal data. The forms were judged by information completeness criteria "conformed" or "not conformed" with standards. Our analysis demonstrate that physicians often are unaware of legal and ethical topics related to constructing informed consent forms. They usually omit crucial information or would use strict technical language. So far, we conclude Hospital Management's Staff definitively has a role in improving informed consent forms by establishing guidelines and promoting ethical issues. Final result could be to deserve stronger patient confidence in medical institutions. PMID- 14552205 TI - [The concept of patient education]. PMID- 14552206 TI - [Praxis approach in educational interventions at the Institute of Nursing Education]. AB - The current development of health awareness in France will not be felt in hospitals and other healthcare organizations in regards to health and health education. Curative and preventive do not seem to be in complete interaction. None-the-less, in the face of a conceptual and political evolution, it is urgent to enlighten, through initial nursing training, future health-care workers towards new functions in the area of education. Therefore in this study we suggest a reflective study on the creation of practical educational competencies. We will be able to demonstrate through this praxiological approach how we will succeed in elaborating two work models: the elaboration of a competency reference pertinent to public health-care the conception of a didactic learning strategy based on a progressiveness concept. PMID- 14552207 TI - [Concepts of health perception and representation: value and limits in the health education domain]. PMID- 14552208 TI - [Psychosocial factors related to the use of psychotropic drugs in elderly persons]. AB - This descriptive study examined psychosocial factors associated with the use of psychotropic medication among older adults living in the community (N = 160). Comparative and descriptive statistical analysis were performed. When compared with non users, psychotropic medication users were older and less formally educated; they were less satisfied with their life and their health; they had greater health problems, reported more depressive symptoms and had more limitations in their daily activities; and they scored lower on sleep satisfaction. Nurses should be aware of the psychotropic medication use among older adults and informed older adults about the risk of addiction and the pernicious effects of long term usage. PMID- 14552209 TI - [Information transfer between health structures. How and to whom is the information transferred?]. AB - The evolution of the French health-care system is based on the development of patient management in the health-care system. The public services associate nurses with alternatives to hospitalization in order to favor the emergence of such facilities. This evolution changes the habits and the needs of professionals. Communication between the nurses is the basis of this function. In order to assure the continuity of health-care, both private and hospital nurses need information. However this is either not the case, or is inadequate to the expectations and needs of the nurses. The motives will be analyzed and some proposals offered. In recording and analyzing liaison records, interesting accounts were revealed which foresees the possibilities of an evolution at the information network level between the hospital and the extra-hospital sector. Some propositions will be put forth in order to assure a greater coherence in the endeavor for health-care continuity. PMID- 14552210 TI - [In the hospital setting, what is the patient's role?]. AB - It is crucial for all medical institutions to maximize the selection of their actors in strict accordance to equality. As medical administrators, were are questioned on the problems linked with compartmentalization. Our investigation has allowed us to confirm that the ensemble of actors, including the patient, regulates compartmentalization, and that all in all it works rather well. However, this regulation has a price. It has an impact on organization, the energy used, not to mention the cumbersome side-effects that would be felt. We have chosen to focus our research on the patient's role on regulation by using his/her testimony. The patient is never passive. Adaptation requires real efforts on the patient's part in order to adapt to this new environment, while at the same time economizing his/her strength. The regulation carried out is often standardized. The patient conforms, but against his/her will. Does the patient's admittance to the hospital make him/her a second-class citizen? Our results lead us to believe that the prevailing views advocate that the patient be at the heart of the system remain an objective. PMID- 14552211 TI - [Health education action in a high-school in suburban Montpellier]. PMID- 14552212 TI - [Health education action in nursing education institutes: experience at the Pontoise Institute]. PMID- 14552213 TI - Nitrous oxide safety. PMID- 14552214 TI - SARS and the dental professional. PMID- 14552215 TI - Dentists can help detect patients at risk of osteoporosis. PMID- 14552216 TI - Acne treatment may cause appearance of discolored gums. PMID- 14552217 TI - Tooth decay a common infectious disease among kids, says CDC. PMID- 14552218 TI - ADA, Academy of Pediatrics: include dental check-up in back-to-school routine. PMID- 14552219 TI - Grant funds program to treat HIV/AIDS patients. PMID- 14552220 TI - Dental hygiene's influence on the marketability and value of a dental practice. PMID- 14552221 TI - Enhancing smiles using microhybrid composite systems. PMID- 14552222 TI - It's all in your contacts! A Class II matrix roundup. PMID- 14552223 TI - Aesthetic ceramic posterior inlays from start to finish. PMID- 14552224 TI - Maxillary first molars and their 4 canals ... the benefits of magnification. PMID- 14552225 TI - Immediate complete maxillary dentures. Aesthetics, fit, and precision. PMID- 14552226 TI - Predictable soft tissue management with radiosurgery. PMID- 14552227 TI - Simplified fixed implant prosthetics. PMID- 14552228 TI - Orthodontics as a prerequisite to implant placement. PMID- 14552229 TI - A conservative aesthetic solution for a single anterior edentulous space. The fiber-reinforced resin fixed partial denture. PMID- 14552230 TI - Stabilizing periodontally compromised teeth with fiber-reinforced composite resin. PMID- 14552232 TI - How to build your dream practice, Part 1. The first steps. PMID- 14552231 TI - Sinusitis as a source of dental pain. AB - Sinusitis is a common medical problem that can occasionally manifest as dental pain. If the patient is experiencing dental pain in the maxillary posterior teeth, then it is appropriate for the dentist to rule out sinusitis as a source of the problem before proceeding with definitive dental treatment. Often there is an obvious odontogenic source of the pain, and this should be resolved first, but in other situations it is difficult to determine the cause of the symptoms. In some patients, the source of the pain is so equivocal that it may be necessary to treat the patient for sinusitis to eliminate this as the source of the dental pain (Table 7). In this process, the dentist has one of 2 options: either refer the patient to a physician or treat the sinusitis. The option chosen regarding patient management is made by the dentist and depends on the particular clinical situation and the dentist's training and experience. PMID- 14552233 TI - Success simply isn't enough.... You can experience significance! PMID- 14552234 TI - Digital radiography and paperless dentistry. PMID- 14552235 TI - Buyers' guide to dental software. PMID- 14552236 TI - Buyers' guide to curing lights. PMID- 14552237 TI - SARS and SAARC: lessons for preparedness. PMID- 14552238 TI - Late weaning: the most significant risk factor in the development of iron deficiency anaemia at 1-2 years of age. AB - BACKGROUND: The global prevalence of iron deficiency anaemia in young children is quite high and children between the ages of 1-2 years are at maximum risk. The complications of anaemia are well known, and side effects may go unnoticed and may have an adverse effect on child's life. Therefore, prevention of anaemia becomes enormously important, and the need to look for parameters and predisposing factors that may lead to iron deficiency anaemia in small children is imperative. This study was designed to determine the association of iron deficiency anaemia with late weaning in 1-2 years of children. METHOD: A case control study was conducted from July 1993-July 1995, at the Community Health Centre (CHC), of the Aga Khan University Hospital, at Karachi, Pakistan. The study included 50 cases and 100 controls. A questionnaire was filled by mothers after taking consent. Data was analyzed by chi-square; t-tests, bivariate analysis and multiple logistic regression. RESULTS: Through bivariate analyses, late weaning, family income, mother's education, the numbers of pregnancies, live births and living children, were found to be statistically significant. These variables were run through a multiple logistic regression model and late weaning was found to be the most significant. 60% of cases and 9% of controls were weaned late (p < 0.001). The mean age of weaning was 7.04 months among cases and 4.46 months among controls (p < 0.001). CONCLUSION: Among all the variables studied, late weaning was the most important predictor of iron deficiency anaemia in 1-2 years of age. PMID- 14552239 TI - Maxillofacial trauma: current practice in management at Pakistan Institute of Medical Sciences. AB - BACKGROUND: This study was carried out to determine the aetiology, pattern and management of maxillofacial injuries at PIMS, Islamabad. METHODS: This descriptive study was conducted at Plastic Surgery Department, PIMS Islamabad from 1st February 1998 to 30th April 2002. All the adult patients presenting with maxillofacial injures were included where as patients less than 12 years of age and only facial lacerations were excluded. Similarly isolated nasal bone fractures were also excluded because these patients were routinely managed by ENT department. Age, sex, presentation, aetiology, associated injuries and treatment modalities undertaken in these patients were recorded. RESULTS: In 164 patients 254 fractures were noted. Most were male (86%), ranging in age from 13-71 years with a male to female ratio of 6:1 respectively. The most frequent (48%) cause noticed was road traffic accidents followed by assault. Mandible was the commonest to be involved in such injuries followed by maxilla. Most of the patients (32%) had associated facial injuries. Various treatment modalities were practiced. CONCLUSION: Maxillofacial fractures should be managed by open reduction and internal fixation as early as possible. PMID- 14552240 TI - Spectrum of mandibular fractures at a tertiary care dental hospital in Lahore. AB - BACKGROUND: Mandible being the only mobile bone of the facial skeleton plays a major role in mastication, speech and deglutition. Among the maxillofacial trauma, mandible is the high risk exponent. Its fractures result in severe loss of function and disfigurement. The increased vehicular accidents, conditions of our roads and other assaults have focused attention in recent years to prevent injuries. METHOD: A descriptive case series of 126 patients with mandibular fractures attending a tertiary referral centre over a six-month period is presented. Data concerning the patients' demographics, aetiology and pattern were obtained and analysed. RESULTS: Approximately 56% of the patients sustained fractures from road traffic accidents and 34% of the fractures occurred in the 21 30 year age range. There was a male preponderance with a male to female ratio of 9:1. Parasymphyseal fractures were the most common followed by those of the condylar and subcondylar regions. CONCLUSIONS: We need to encourage further research work in this field to provide better record keeping and quality of services to the injured people. Amendments in the traffic rules and legislation about seat-belt usage and traditional kite flying to reduce frequency of fractures are required. PMID- 14552241 TI - Profile of men suffering from sexually transmitted infections in Pakistan. AB - BACKGROUND: To evolve effective prevention and efficient treatment strategies for sexually transmitted infections (STIs) in a country, comprehensive understanding of the prevalent STI and their modes of transmission is needed. The aim of this present study was to generate such data for Pakistan. METHOD: The study was conducted between June 1999 and September 1999 in four provincial capitals. In each city, data were collected from one or more teaching hospitals and a number of general practitioners. During this period, 465 men suffering from STIs were interviewed. RESULTS: The mean age of study population was 31.9 +/- 8.6 years, 60% were married, 58.3% urbanites, 81.7% living with their families and 83.3% smokers. Only 10.5% were drug addicts. Out of 465 cases, 27.5% cases had gonorrhoea, 31.6% syphilis, 17.2% chancroid, 18.3% herpes, 5.2% chlamydial infections while only one case (0.2%) was HIV positive. Most men (55%) acquired the infection heterosexually, 11.6% through homosexuality, 18.4% through bisexual relations and 2 men (0.5%) reported bestiality. Among 78.1% of those contracting the infection heterosexually, the source of infection was a sex worker. None acquired infection through his wife. The knowledge about STIs was very poor. Wrong notions were prevalent. CONCLUSIONS: The pattern of STIs is different from developed countries. Those acquiring STIs in Pakistan were older, married and living with their families whereas in developed countries most men acquiring STIs are younger, unmarried and living alone. PMID- 14552242 TI - Analysis of urine cytology at a community hospital. AB - BACKGROUND: To review the pattern of urine cytology in our setting of a community hospital with age and gender distribution. Also to discuss the importance of this safe and inexpensive tool for diagnosing and screening urothelial cancers. We desire to encourage more wider and confident use of urine cytology in the routine practice. METHOD: We reviewed all the consecutive urine cytology specimens received at our institution during time frame of one year. The cytological diagnosis were grouped in four categories; 1: negative, benign or reactive cases; 2: cases showing acute inflammatory changes; 3: inconclusive cases and 4: malignant cases. Only the positive malignant cases were followed histologically to determine the accuracy and efficiency of urine cytological examination for positive cases. RESULTS: A total of 1957 consecutive urine cytology specimens were reviewed. Majority of the cases were negative for tumour (67.19%). The most common diagnostic category was acute inflammatory case (17.3%) while inconclusive cases constituted 11.39% of total. There were 23 (1.18%) positive cases that were diagnosed malignant on urine cytology. These 23 malignant cases were followed histologically and 22 (95.6%) cases turned out to be malignant. CONCLUSION: Urine cytological examination is an efficient tool that has a good diagnostic yield in detecting malignant urothelial lesions. It should be used routinely for diagnosing, screening and for follow-up of all malignant urothelial lesions. PMID- 14552243 TI - Bacterial aetiology of osteomyelitis cases at four hospitals of Lahore. AB - BACKGROUND: The conclusive diagnosis of osteomyelitis requires isolation of pathogen in aspirate from bone lesion, bone debridement and blood culture. The present research was undertaken to study the microbiological pattern of cases of osteomyelitis reporting to four hospitals in Lahore. METHOD: One hundred and fifty patients of osteomyelitis were selected from outpatient departments and Orthopaedic wards of Lahore General Hospital, Sir Ganga Ram Hospital, Services Hospital and Mayo Hospital, Lahore. Specimens of pus from bone, blood and bone debridement were collected. All samples were inoculated onto two Blood Agar and one MacConkey agar plates. One Blood Agar plate was incubated anaerobically for 48 hours and the other two plates aerobically for 24 hours. Smears were made from samples and stained by the Gram's stain. The colonies obtained were processed according to the technique of Mackie and MacCartney. RESULTS: The commonest isolates belonged to the Enterobacteriaceae (32.8%), followed by Staphylococcus aureus (29.5%). Pseudomonas aeruginosa (15.5%), anaerobes (2.6%) and miscellaneous (19.3%). Five (2.7%) anaerobic bacteria were isolated. Anaerobic bacteria were peptostreptococci, peptococci and bacteroides either alone or as a mixed infection. CONCLUSION: The present study highlights the importance of microbiological examination of bone in cases of osteomyelitis. Different types of bacteria either alone or as a mixed infection could be the causative agent(s). PMID- 14552244 TI - Frequency of intestinal parasitic infestation in children of 5-12 years of age in Abbottabad. AB - BACKGROUND: Worm infestation is a major problem in children from developing countries due to bad hygienic conditions. It produces nutritional deficiencies and anaemia in children, especially when hookworm infestation is present. METHODS: This cross-sectional study deals with investigation of the frequency of intestinal parasitic infestation in children between the ages 5-12 years. A total of 283 subjects were tested and screened for different intestinal parasites at of Department of Physiology, Ayub Medical College, Abbottabad. Negative cases were re-examined and if found free of intestinal pathogenic parasites were labelled as negative. RESULTS: Of the 283 children examined, 230 tested positive for various intestinal parasites. The frequency of helminthic infestation was found to be above 81%. There were 8 different species of helminths and protozoa found in the specimens. By far the highest frequency of 48% was noted for Ascaris lumbricoides while 6.9% (16 cases) of the specimens examined had mixed infestation. The mean Haemoglobin (Hb) level was found to be 9.82 g/dl in males and 9.0 g/dl in females. Virtually no Hookworm infestation was found which may be the reason of not so low Hb level of the subjects. CONCLUSIONS: A very high percentage (81%) of children from suburbs of Abbottabad have intestinal worm infestation and majority of them (48% of positive cases) have Ascaris lumbricoides. Children were not very severely anaemic because of virtually no hook worm cases. PMID- 14552245 TI - Use of folk remedies among patients in Karachi Pakistan. AB - BACKGROUND: The concept that food is medicine is being practiced in certain parts of the world, with positive outcomes on health of the population. We have such practice in Pakistan but it needs to be brought in line with the available scientific evidence. METHODS: The study was conducted on 270 patients, visiting the Family Practice Center, the Aga Khan University, Karachi. A questionnaire was used to collect information on the demographic profile, and the use of folk remedies for medicinal uses. RESULTS: Substantial use of folk remedies for different medical conditions has been documented. The remedies included cinnamon, ginger, cloves, cordimon, sesame oil, poppy seeds, honey, lemon, table salt, eggs and curd. The medical conditions in which folk remedies are used in respondents' view, include conditions such as common cold, cough and flu to more serious conditions such as asthma, jaundice and heat stroke. CONCLUSIONS: We have found a substantial use of folk remedies for treatment of medical conditions. There is a need to organize their use on scientific lines. PMID- 14552246 TI - Comparison of store and forward method of teledermatology with face-to-face consultation. AB - BACKGROUND: To compare the accuracy of store and forward method of teledermatology with the traditional face-to-face consultation. METHODS: The comparison was done between Institute of Dermatology King Edward Medical College Lahore that served as teledermatology center and Dermatology Department of Pakistan Institute of Medical Sciences Islamabad from where patients were selected. Telmedpak provided the technical support. Thirty three patients were selected from outpatient department of PIMS and images were taken using a digital camera. Images were stored in computer and were sent to Institute of Dermatology via e-mail for Teleconsultation along with a short history and examination findings. Diagnosis of consultant after face-to-face consultation was then compared with the image based diagnosis that is after teleconsultation. RESULTS: In 81% of the cases the diagnosis on face-to-face consultation was same (p < 0.05) while in 18% of the cases the two diagnoses differed. In 9% (n = 3) of the cases, image resolution was not good but out of these three, diagnosis was same in two and differed in one case. CONCLUSION: This study concludes that store and forward method of teledermatology is reliable and can provide a means of increasing access to dermatological care in rural and under-served areas. PMID- 14552247 TI - Sricturoplasty in tuberculous small bowel strictures. AB - BACKGROUND: This study was conducted at DHQ Hospital Hangu, Babul Madina Medical Centre & Shifa Medical Centre Hangu from March 1995 to February 2001 to evaluate the presentation of tuberculous small bowel strictures and assess the usefulness and applicability of stricturoplasty to these lesions. METHODS: Ten patients were included in the study in whom the diagnosis of tuberculous small bowel strictures was made on laparotomy and histopathological examination of the tissues taken from the strictures. Heinki/Mikulikz's type stricturoplasty was done in all the cases. The patients were kept on antitubercular drugs and were followed for assessment of outcome. RESULTS: Seventy percent of the patients' presented/operated upon in emergency. Age ranged from twenty to forty years with mean age 31.5 years. Male to female ratio was 6:4. Weight loss and malnutrition were the most common associated features. Terminal ileum alone was involved in 50% of the cases. The strictures were solitary in six and multiple in four cases. The lesions were acute in four and chronic in six patients. Stricturoplasty worked well both in acute and chronic cases. CONCLUSION: Stricturoplasty is safe, simple, and easy procedure particularly suitable at poorly equipped and under staffed district hospitals. PMID- 14552248 TI - Renal cortical necrosis: a case series of nine patients & review of literature. AB - BACKGROUND: The purpose of this study is to review cases of renal cortical necrosis (RCN) which we came across during a period of five years, confirmed on needle biopsy specimens, with literature available so far. METHODS: All renal biopsy records between January 1996 to December 2001 at the Aga Khan University Hospital, Karachi, Pakistan were reviewed to identify patients with histologically proven renal cortical necrosis. Seven patients were also referred by the department of Nephrology, Lady Reading Hospital, Peshawar, Pakistan. Patients' medical records were reviewed for demographic, clinical, and follow up data. RESULTS: There were total of nine patients identified with renal cortical necrosis. The mean age was 26.21 +/- 13.01 year. Five cases (55.5%) were caused by obstetric complications. The most common histology type of RCN was patchy cortical necrosis in seven patients (77.8%). Three patients were alive at last follow up; remaining patients were lost to follow up. CONCLUSION: Obstetric complications remain the leading cause of renal cortical necrosis in our setting. The recognition of this entity and value of renal biopsy in cases of acute renal failure needs to be emphasized as this helps in establishing diagnosis, to assess prognosis and to aid in selection of a rational approach to therapy. PMID- 14552250 TI - Analysis of maternal mortality in a tertiary care hospital to determine causes and preventable factors. AB - BACKGROUND: All pregnant women are at risk of obstetrical complications and most of these occur during labour and delivery that lead to maternal death. In our setup maternal mortality is seriously under estimated. Safe motherhood as a priority for action can not be identified with out properly assessing maternal mortality. The objectives of this study were to determine causes and preventable factors responsible for maternal mortality. METHODS: This study was conducted in Gynaecology 'C' unit of Ayub Teaching Hospital, Abbottabad, from January 2000 to December 2001. Patients were admitted through emergency obstetrics care unit and Gynaecology out patient department. Patients' demographic record including age, parity, education, socio-economic status along with antenatal care record, level of care and distance from hospital were noted. Causative factors leading to maternal death and contribution factors evaluated. All this information was collected from patients' records. RESULTS: Twenty-six maternal deaths were recorded during study period. The major causative factors were haemorrhage 9 (34.6%), eclampsia 8 (30%), sepsis 5 (19.2%), anaesthetic complications 3 (11.5%) and hepatic encephalopathy 1 (3.8%). Maternal mortality ratio was 12.7/1000 live births (26/2040). The age range was between 18-42 years. There were 16 (61.5%) patients in > 30 years age group. Most of them (69%) were grand multiparas (Parity > 5). Education, antenatal booking and socio-economic status were poor. The distance from hospital was between 10 and 100 KM. The level of care available at nearest health facility was estimated, 40% were attended by traditional birth attendants, 33% by lady health visitors, 10% by doctors and to 17% no level of care was available. CONCLUSION: Obstetrical haemorrhage and hypertensive disorders are still major causes of maternal deaths. Most maternal deaths are preventable. The provision of skilled care and timely management of complications can lower maternal mortality in our setup. PMID- 14552249 TI - Prognosis of gestational choriocarcinoma at Khyber Teaching Hospital Peshawar. AB - BACKGROUND: Choriocarcinoma is a highly malignant tumour which originates in developing trophoblast of pregnancy, most commonly following molar pregnancy. It is a potentially fatal disease, but current management protocols have turned the prognosis highly favourable. METHODS: This study was done on patients with gestational choriocarcinoma presenting to Gynae-B unit of Khyber Teaching Hospital Peshawar, between May, 1996 to December, 1997, diagnosed on the basis of clinical course and elevated level of HCG. Metastatic evaluation of the disease was done to assign different risk groups to the patients before selecting appropriate chemotherapy regimen for each patient. Results of the therapy were monitored by serial estimation of HCG levels. RESULTS: During this period 5 patients of choriocarcinoma were treated. In 2 (40%) cases choriocarcinoma developed after molar pregnancy whereas in 3 (60%) cases antecedent pregnancy resulted in spontaneous abortion. Four (80%) patients were from poor socioeconomic class, 3 (60%) were above 39 years of age and 4 (80%) were multiparous. Two patients (40%) were medium risk and 3 (60%) were high risk cases. There was no patient with low risk disease. EMA-CO (Etoposide, Methotrexate, Actinomycin-D, Cytocine, Oncovine) regimen was administered to all patients. Maximum number of cycles of chemotherapy given was 8. Only one patient developed drug resistance. Overall cure rate was 80% (4 patients survived out of 5 at two years' follow-up). CONCLUSION: Prognosis of gestational choriocarcinoma is favourable provided the appropriate therapy is administered early in the course of disease. Provision of free medical care should be considered for these patients to save their lives. PMID- 14552251 TI - Hepatitis C seropositivity among chronic liver disease patients in Hazara, Pakistan. AB - BACKGROUND: Hepatitis C is rapidly emerging as a major health problem in developing countries including Pakistan. The present study was conducted to document the frequency of Hepatitis C seropositive individuals reporting for hepatitis testing at a referral laboratory. METHODS: Serum samples were collected from 614 people (436 males and 178 females) referred for chronic liver disease from all parts of Hazara division during the period July 2000 to July 2002. Hepatitis tests were performed by DOT immuno-chromatographic method for anti HCV antibodies. RESULTS: A total of 251 (40.8%) sera tested positive for anti HCV antibodies, including 184 males (73.3%) and 67 females (26.7%). Of 436 males, 184 (42.2%) tested positive, while among females, 67/178 (37.6%) tested positive; this difference is not statistically significant. However the male/female ratio referred for testing was 2.4:1. CONCLUSION: There is a high frequency of HCV seropositive individuals of both sexes among patients referred for chronic liver disease. The frequencies obtained for Hazara division compare well with figures from other parts of Pakistan as well as developing countries. The male/female ratio could simply be a reflection of more males coming for treatment and testing in our setting. PMID- 14552252 TI - Impact of class attendance upon examination results of students in basic medical sciences. AB - BACKGROUND: A number of studies have shown the impact of attending the classes during the session on examination results of students. Students with good lecture attendance show good results while those with poor lecture attendance are at risk for poor performance in the examinations. In this study we wanted to test this in students of basic medical sciences in our set up. METHODS: All the students in Basic Medical Sciences of Gomal Medical College, D.I. Khan, Pakistan, during the session 1999-2000 were enrolled for this study. Performance of these students in one of their three annual term tests, which covered duration of studies for four months, from February 2000 to May 2000, was analyzed. Students were grouped into two, Group-A with > or = 75% attendance and Group-B with < 75% attendance. The frequency of failure in the two groups was compared using the chi square test. RESULTS: Results of the term test showed that the percentage of failure in students of Group-B with less than 75%, attendance was significantly higher when compared to Group-A with equal or more than 75% attendance. CONCLUSION: Our results supported the previous studies showing that attendance in the classes during teaching sessions had a direct impact on performance of students in the examinations. Students with good attendance show good results while those with poor attendance are at risk for poor performance during examinations in basic medical sciences. PMID- 14552253 TI - Accuracy of ultrasound in the diagnosis of upper abdominal pain. AB - BACKGROUND: Upper abdominal pain is one of the commonest complaints in patients seeking medical advice, and in majority of the cases the aetiology is simple and treatable. Ultrasonography is hazard free and cost effective investigation. This study was designed to evaluate the accuracy of ultrasound in the diagnosis of upper abdominal pain. METHODS: Ultrasonography using 3.5 MHz convex transducer was performed in patients presenting with upper abdominal pain. RESULTS: Five hundred consecutive cases with upper abdominal pain underwent ultrasonic examination. Positive findings were there in 248 patients while 252 had normal studies. CONCLUSION: This study showed that ultrasound is an easy and quick investigation in the diagnosis of upper abdominal pain with slightly less than 50% accuracy. PMID- 14552254 TI - Leiomyoma of the vagina. AB - A vaginal leiomyoma is normally a benign smooth muscle tumour in the vagina. These tumours are extremely rare and the aetiology is unknown. The imaging findings are those of a nonspecific well-defined enhancing soft tissue mass centred on the vagina. The lesion may be very large, but is usually under 6 cm in size. Patients are asymptomatic in the early stages. Symptoms arise with the growth of tumour mainly due to compression. Most leiomyomas are not diagnosed clinically but only on histological examination. A case of primary leiomyoma of vagina is presented here. PMID- 14552255 TI - Cancer ovary, present and future of management. AB - Ovarian cancer is very important malignancy of woman as far as its incidence and mortality is concerned. Although majority presents at late stage but still there is a reasonable response to currently available chemotherapy drugs and their use in multimodality setting. Besides good response to chemotherapy drugs, majority have recurrence. So there is a need for new drugs, new trends and their combinations. All these issues will be discussed in this review article. PMID- 14552258 TI - It's not just peanuts. PMID- 14552259 TI - Diabetes--a review. PMID- 14552260 TI - Recognizing teenage depression. PMID- 14552261 TI - Danger: mercury is hazardous to our health. PMID- 14552262 TI - Informed consent: adolescent minors, surrogate decision-making, and the school nurse. PMID- 14552263 TI - PANDAS syndrome in the school setting. PMID- 14552264 TI - The impact of FERPA and HIPAA on privacy protections for health information at school. PMID- 14552265 TI - Community approaches to a national problem: oral health. Part 1. PMID- 14552266 TI - 'Helping the student with diabetes succeed' a new resource for effective diabetes management in school. PMID- 14552267 TI - The impact of culture on health and wellness: culture counts. PMID- 14552268 TI - For those who care. PMID- 14552269 TI - Crimes come in many forms. PMID- 14552270 TI - The giving of care. PMID- 14552271 TI - Caregiving and the physician ... professional and personal perspectives. PMID- 14552272 TI - The many phases of caregiving. PMID- 14552273 TI - Pain, pain everywhere ... almost. PMID- 14552274 TI - When the physician needs care. PMID- 14552275 TI - Elder abuse. PMID- 14552277 TI - Error in medicine. PMID- 14552276 TI - Taking care of Maryland's older adults. PMID- 14552278 TI - Extra embryos. PMID- 14552279 TI - Crossing over the barriers of prejudice. PMID- 14552280 TI - West Nile virus. PMID- 14552281 TI - Double trouble. PMID- 14552282 TI - Important notice regarding Aetna class action settlement. PMID- 14552283 TI - Positive partners: consumer involvement in HIV care. PMID- 14552284 TI - So soft drinks contribute to obesity? PMID- 14552286 TI - Dealing with head lice--how have times changed? PMID- 14552285 TI - . . . And pediatricians speak out on the childhood obesity epidemic. PMID- 14552287 TI - FDA approves growth hormone for short children. PMID- 14552288 TI - Does spending more time in child care make children more assertive and aggressive? PMID- 14552289 TI - Recalls: extended recall of Cosco Arriva and Turnabout infant car seats/carriers. PMID- 14552290 TI - Increase of atmospheric CO2: response patterns of a simple terrestrial man-made ecosystem. AB - Simple models of terrestrial ecosystems with a limited number of components are an efficient tool to study the main laws of functioning of populations, including microbial ones, and their communities, as components of natural ecosystems, under variable environmental conditions. Among other factors are the increase of carbon dioxide in the atmosphere and limitation of plants' growth by biogenic elements. The main types of ecosystems' responses to changes in environmental conditions (a change in CO2 concentration) have been demonstrated in a "plants-rhizospheric microorganisms-artificial soil" simple experimental system. The mathematical model of interactions between plants and microorganisms under normal and elevated atmospheric CO2 and limitation by nutrients (nitrogen and phosphorus) yielded a qualitative agreement between calculated and experimental values of limiting substances concentrations and release rates of exudates. PMID- 14552291 TI - Biology and physiopathology of angiogenesis: the 1997 Philippe Laudat Conference. PMID- 14552292 TI - New challenges for ethics consultation: combining feminism, multiculturalism, and caring. PMID- 14552293 TI - Ethics consultation in a culturally diverse society. PMID- 14552294 TI - The practice of a clinical ethics consultant. PMID- 14552295 TI - The feminist health care ethics consultant as architect and advocate. PMID- 14552296 TI - Contrasting role morality and professional morality: implications for practice. AB - The notion of role morality suggests individuals may adopt a different morality depending on the roles they undertake. Investigating role morality is important, since the mentality of role morality may allow agents to believe they can abdicate moral responsibility when acting in a role. This is particularly significant in the literature dealing with professional morality where professionals, because of their special status, may find themselves at odds with their best moral judgments. Here I tell four stories and draw out some distinctions. I conclude that role morality is a genuine and useful distinction. However, I suggest that the purported distinction between role morality and professional morality is over-determined. Therefore, alleged conflicts between the demands of role and profession (such as the different pressures on Pinto designers as employees and as engineers) are not conflicts between different kinds of demands, but rather conflicts arising from divergent roles that most workers will encounter regularly. Another analytical perspective is to look at moral choices at work in terms of power and the ability to bring about change. Finally, I draw the implication that we should stress moral awareness at a fairly abstract level for all employees and reinforce the moral primacy of individual choice. PMID- 14552297 TI - Gamete donation and parental responsibility. AB - Unlike surrogacy and cloning, reproduction via gamete donation is widely assumed to be morally unproblematic. Recently, a number of authors have argued that this assumption is mistaken: gamete donors, they claim, have parental responsibilities that they typically treat too lightly. In this paper I argue that the 'parental neglect' case against gamete donation fails. I begin by examining and rejecting the view that gamete donors have parental responsibilities; I claim that none of the current accounts of parenthood provides good reason for ascribing parenthood to gamete donors. I then argue that even if gamete donors do have parental responsibilities for 'their' children, it is not clear that they treat these responsibilities too lightly. I conclude the paper by examining the wider question of just what kind of responsibilities gamete donors might have towards the children that they have a role in creating. PMID- 14552298 TI - Children as guinea pigs: historical perspective. AB - Experimentation involving children is not a new phenomenon. Children have been used as research subjects in a diverse set of experiments, including the trials of new vaccines and sera, in efforts to understand normal pediatric anatomy and physiology and in the development of new drugs and procedures. Concern about child participants in research is also not a new development. For more than a century, critics of medical research have called attention to the fact that children and other vulnerable populations--pregnant women, prisoners, the mentally ill--have too often served as the unwitting and unwilling subjects of medical experiments. This paper looks at several early cases in which children participated, including the first trial of cowpox vaccine, the first human trial of rabies vaccine, and the first treatment of Listerian wound antisepsis. The history of concern for children, especially institutionalized children, in medical research is considered along with the development of regulations or guidelines, including the Declaration of Helsinki (1964). PMID- 14552299 TI - Pediatric ethics and early-phase childhood cancer research: conflicted goals and the prospect of benefit. PMID- 14552300 TI - A new world order for human experiments. PMID- 14552301 TI - Overwhelming casualties: medical ethics in a time of terror. PMID- 14552302 TI - The use of experimental drugs during national crisis. PMID- 14552303 TI - Research challenges for clinical investigators in rheumatology. PMID- 14552304 TI - Internet research: an opportunity to revisit classic ethical problems in behavioral research. AB - The Internet offers many new opportunities for behavioral researchers to conduct quantitative and qualitative research. Although the ethical guidelines of the American Psychological Association generalize, in part, to research conducted through the Internet, several matters related to Internet research require further analysis. This article reviews several fundamental ethical issues related to Internet research, namely the preservation of privacy, the issuance of informed consent, the use of deception and false feedback, and research methods. In essence, the Internet offers unique challenges to behavioral researchers. Among these are the need to better define the distinction between private and public behavior performed through the Internet, ensure mechanisms for obtaining valid informed consent from participants and performing debriefing exercises, and verify the validity of data collected through the Internet. PMID- 14552305 TI - Using of disodium clodronate on DSO patients. PMID- 14552306 TI - Voluntary assent in biomedical research with adolescents: a comparison of parent and adolescent views. AB - An informed consent and voluntary assent in biomedical research with adolescents is contingent on a variety of factors, including adolescent and parent perceptions of research risk, benefit, and decision-making autonomy. Thirty-seven adolescents with asthma and their parents evaluated a high or low aversion form of a pediatric asthma research vignette and provided an enrollment decision; their perceptions of family influence over the participation decision; and evaluations of risk, aversion, benefit, and burden of study procedures. Adolescents and their parents agreed on research participation decisions 74% of the time, yet both claimed ultimate responsibility for the participation decision. Both rated most study procedures as significantly more aversive than risky. Parents were more likely to rate aspects of the hypothetical study as beneficial and to provide higher risk ratings for procedures. Disagreements concerning research participation decisions and decision-making autonomy have implications for the exercise of voluntary assent in biomedical research. PMID- 14552307 TI - A "primary" antiphospholipid syndrome? PMID- 14552310 TI - The ethical ideologies of psychologists and physicians: a preliminary comparison. AB - The ethical ideologies of psychologists (who provide health services) and physicians were compared using the Ethics Position Questionnaire. The findings reveal that psychologists tend to be less relativistic than physicians. Further, we explored the degree to which physicians and psychologists report being influenced by a variety of factors (e.g., family views) in their ethical decision making. Psychologists were more influenced by their code of ethics and less influenced by family views, religious background, and peer attitudes than were physicians. We argue that these differences reflect the varied professional cultures in which practitioners are trained and socialized. PMID- 14552308 TI - Auranofin improves outcome in early rheumatoid arthritis. Results from a 2-year, double blind placebo controlled study. AB - The effect of early initiation of auranofin (AF) therapy on outcome measures was studied in a controlled 24-month double blind trial in 138 patients with early rheumatoid arthritis (RA) using an intent to treat approach. Patients were randomized to AF or placebo but in case of insufficient effect or intolerable adverse events, they switched to open disease modifying antirheumatic drug therapy. Patients who started AF fared significantly better in improved joint swelling. Stanford Health Assessment Questionnaire index, Keitel functional test, and mental depression, and furthermore, radiologic progression was significantly retarded. Our results support a disease modifying beneficial effect of AF in early active RA.se PMID- 14552311 TI - An outbreak of Salmonella enterica serovar typhimurium (S. typhimurium) phage type 135a in the Greater Murray. PMID- 14552312 TI - Intellectual freedom and editorial responsibilities within the context of controversial research. AB - The primary purpose of this article is to explore the limits that an agent, such as the government or the American Psychological Association, may place on one's right to pursue a program of research or to share the findings of a research project. The primary argument that evolves here is that researchers' rights to pursue an interesting hypothesis, and their freedom of expression, are conditional. The author examines the potential pragmatic and epistemological barriers to a program of research and the responsibilities that researchers bear to address epistemological and nonepistemological matters while reviewing the implications of their work. PMID- 14552313 TI - Task demands, task interest, and task performance: implications for human subjects research and practicing what we preach. AB - Through the continuous investigation of humans in organizations, we have learned much about motivation, attitudes, and performance. For example, Yukl and others have helped increase our understanding of influence tactics and the effect they have on the performance of subordinates, supervisors, and peers. Some tactics (and combinations of tactics) lead to resistance, some lead to compliance, and some lead to commitment. In this study, we raise the question of whether or not we incorporate our knowledge of these research findings into the design, implementation, and interpretation of our own research studies that require the participation of human subjects. In a survey of 134 subjects from a previous social science study, we found that performance varied across the sample, consistent with the concepts of resistance, compliance, and commitment. In addition, the variance in performance could be explained, in part, by task interest and perceived task demands. Implications are discussed. PMID- 14552314 TI - Dual relationship activities: principal component analysis of counselors' attitudes. AB - The British Columbian Members of the Canadian Guidance and Counselling Association were surveyed to explore their attitudes regarding dual relationships. Of 529 deliverable surveys, 206 usable returns yielded a response rate of 39%. The survey instrument collected data regarding respondents' characteristics and ethicality ratings of 39 dual relationship activity items. An exploratory principal components analysis was performed on responses, resulting in a 4-factor equation, which accounted for 44% of the total variance. The results suggest that, although conceptual considerations of dual relationship typology do underlay the resultant factors, the relative ethicality of each item is also influential. PMID- 14552315 TI - What the lions say. PMID- 14552316 TI - A new approach using intraoperative ultrasound imaging for the localization and removal of multiple foreign bodies in the neck. AB - An innovative method of using ultrasound imaging intraoperatively is reported. Multiple radiopaque and radiolucent foreign bodies within soft tissues were localized in theatre using high resolution ultrasonography. Depth of injury and vessel status were also ultrasonically assessed. Surgical removal of all foreign bodies was aided by intraoperative ultrasound guidance. In addition, this new approach of using intraoperative ultrasonography was pivotal in confirming the removal of all foreign bodies from the wound at the time of the operation. PMID- 14552317 TI - National Institutes of Health Recombinant DNA Advisory Committee (RAC). 91st meeting of the NIH RAC, Bethesda Marriott Hotel, Bethesda, Maryland, June 18-19, 20003. PMID- 14552318 TI - Frequent Ki-ras mutations in gastric tumors of the MSI phenotype. PMID- 14552319 TI - Albumin for refractory ascites. PMID- 14552320 TI - The interferon sensitivity determining region in the era of combination therapies and the rational use of such therapies for patients with HCV genotype 1b infection. PMID- 14552321 TI - Discussion on genotype and phenotype correlations in patients with cystic fibrosis and pancreatitis. PMID- 14552322 TI - Effect of long-term lamivudine therapy on histological outcome in chronic hepatitis B. PMID- 14552323 TI - Neuroendocrine cell secretory granules are not bacteria in inflammatory bowel disease. PMID- 14552324 TI - Screening for celiac disease in patients with chronic liver disease. PMID- 14552325 TI - Crohn's disease remission on bupropion. PMID- 14552326 TI - Novel protein targeted therapy of metastatic melanoma. AB - Metastatic melanoma patients have a dismal prognosis with poor responsiveness to chemotherapy, radiation therapy and current immunotherapy regimens and a median survival of less than six months. Novel therapies directed at melanoma-selective molecular targets are urgently needed. Based on the frequent constitutive activation of the mitogen-activated protein kinase (MAPK) signaling pathway in malignant melanomas and the selective inhibition of MAPK signaling by anthrax lethal factor which proteolytically cleaves MAPK kinases, anthrax lethal toxin may be a useful agent for patients with metastatic melanoma. Anthrax lethal toxin consists of two proteins--protective antigen and lethal factor. These two proteins have been separately produced in good yields and in high purity. The three-dimensional structures of these proteins have been solved, and their molecular mechanisms of cell binding and action determined. Preclinical studies with anthrax lethal toxin show sensitivity of malignant melanoma cell lines in tissue culture and anti-tumor efficacy in melanoma xenograft models. Additional studies to define the maximal tolerated doses and dose-limiting toxicity of anthrax lethal toxin in rodent and primate models should pave the way for phase I studies testing the efficacy of the anthrax lethal toxin in patients with metastatic melanoma. PMID- 14552327 TI - COX-2 inhibition in esophagitis, Barrett's esophagus and esophageal cancer. AB - There is extensive evidence that cyclooxygenase-2 (COX-2) plays a significant role in the process of carcinogenesis in different tumors. Although most of these evidences derive from studies in colorectal cancer, data obtained from recent studies strongly suggest that COX-2 might play an important role in the neoplastic transformation of esophageal epithelium. NSAIDs use is associated with a reduction of the risk of developing esophageal cancer, including adenocarcinoma. Up-regulation of COX-2 has been reported in different stages of the carcinogenic sequence leading to esophageal adenocarcinoma. Treatment with selective COX-2 inhibitors has been shown to reduce the damage induced by acid and pepsin in the esophageal mucosa of rabbits, the incidence of tumors in an animal model of esophageal adenocarcinoma and to decrease proliferation and induce apoptosis in both Barrett's epithelial and adenocarcinoma cells. The first clinical study has shown that selective inhibition of COX-2 is followed by a significant decrease of cell proliferation in human Barrett's metaplasia. Clinical trials have begun in order to assess the efficacy of selective COX-2 inhibitors to prevent the progression of Barrett's esophagus to adenocarcinoma. Bile salts and acid are likely to early induce COX-2 in this sequence, although other factors, such as proinflammatory cytokines, inducible nitric oxide synthase and growth factors such as TGF-beta, are potential COX-2 inducers in the esophagus. Further studies are necessary in order to better understand factors involved in COX-2 up-regulation and mechanisms of COX-2 associated tumorigenesis in the esophagus. PMID- 14552328 TI - An algorithm for the clinical assessment of nutritional status in hospitalized patients. AB - Upon admission to hospital, 30-50 % of patients either are or become malnourished. There is no generally accepted definition of malnutrition or guidelines on the best way to establish nutritional status. We consider it self evident that the nursing staff have an important role in screening patients at risk of malnutrition on admission and thereafter at regular times. This is why we developed the nursing nutritional screening form (NNSF). The NNSF was tested by nurses, dietitians and clinicians, in pairs, to establish the extent of agreement in two phases on sixty-nine and forty patients. Later, the form was used in practice by nursing staff on five wards (334 patients). Based on the results of the NNSF, patients were referred to a dietitian. The dietitian established whether the patient was indeed at risk, or was actually malnourished, using a complete nutritional history. The degree of concurrence within pairs was reasonable to good. The same applied to the concurrence between nursing staff and dietitians, but concurrence between clinicians and nursing staff was less. In total, 334 patients were screened and sixty-nine of them were referred to the dietitian. It was established that 86 % of the referred patients were potentially at risk of malnutrition or were malnourished. Without the NNSF, 39 % (n 27) of the patients referred to the dietitian would not have been referred, or would have been referred much later. The NNSF makes it possible for nurses to detect malnourished patients or patients at risk of malnutrition at an early stage of their hospitalization. PMID- 14552330 TI - Some insights into the derivation and early uses of the word 'probiotic'. PMID- 14552329 TI - Relationship between body composition and blood pressure in Bahraini adolescents. AB - The objective of the present study was to examine the relationship between body composition and blood pressure (BP) in Bahraini adolescents. A sample of 504 Bahraini schoolchildren aged 12-17 years (249 boys and 255 girls) was selected using a multi-stage stratified sampling procedure. BP measurements were performed on the students. Anthropometric data including weight, height, waist circumference (WC), hip circumference, and triceps, subscapular and medial calf skinfold thicknesses were also collected. BMI, percentage body fat, waist:hip (WHR), and subscapular:triceps skinfold ratio were calculated. Mean systolic BP and mean diastolic BP were higher in males than in females. Weight and height in boys and weight only in girls were significantly associated with systolic BP independent of age or percentage fat. Nearly 14 % of the adolescents were classified as having high BP. BMI and percentage body fat were significantly and positively associated with the risk of having high BP in the boys and girls. Adolescents with high WHR or WC, as indicators for central obesity, tended to have higher BP values. The results from the present study indicate that obesity influences the BP of Bahraini adolescents and that simple anthropometric measurements such as WHR and WC are useful in identifying children at risk of developing high BP. These findings together with the known tracking of BP from adolescence into adulthood underline the importance of establishing intervention programmes in order to prevent the development of childhood and adolescent obesity. PMID- 14552331 TI - Gene Symbol: GLDC. Disease: NKH glycine encephalopathy. PMID- 14552332 TI - Gene symbol: M19489, CYP17. Disease: 17-alpha-hydroxylase/17,20-lyase deficiency. PMID- 14552333 TI - Gene symbol: CYp17, M19489. Disease: 17-alpha-hydroxylase/17,20-lyase deficiency. PMID- 14552334 TI - WHO acts on road safety to reverse accident trends. Traffic accidents kill 1.26 million people each year; 2nd leading cause of death among those aged 15-29. PMID- 14552335 TI - Binge drinking on rise in UK and elsewhere. Government report shows increases in alcohol consumption, cirrhosis, and premature deaths. PMID- 14552336 TI - Indian firm caught marketing anticancer drug for fertility. PMID- 14552337 TI - Decade of pain control and research gets into gear in USA. PMID- 14552338 TI - EC urges reporting of occupational diseases. PMID- 14552339 TI - Information transfer from the USA and Europe to Latin America: current need and suitable methods. PMID- 14552340 TI - Draft guidelines for radiopharmacy. PMID- 14552341 TI - Disease management: a leap of faith to lower-cost, higher-quality health care. AB - With managed care's promise to reduce costs and improve quality waning, employers and health plans are exploring more targeted ways to control rapidly rising health costs. Disease management programs, which focus on patients with chronic conditions such as asthma and diabetes, are growing in popularity, according to findings from the Center for Studying Health System Change's (HSC) 2002-03 site visits to 12 nationally representative communities. In addition to condition based disease management programs, some health plans and employers are using intensive case management services to coordinate care for high-risk patients with potentially costly and complex medical conditions. Despite high expectations, evidence of both disease management and case management programs' success in controlling costs and improving quality remains limited. PMID- 14552342 TI - Man's best friend. PMID- 14552344 TI - Facing harsh realities--in space, air, and war. PMID- 14552343 TI - Steady course for civil space in 2002. PMID- 14552345 TI - Robonauts join "spacewalk squad". PMID- 14552346 TI - A dynamic model of stress and sustained attention. AB - This paper examines the effects of stress on sustained attention. With recognition of the task itself as the major source of cognitive stress, a dynamic model is presented that addresses the effects of stress on vigilance and, potentially, a wide variety of attention performance tasks. PMID- 14552347 TI - Personality and coping strategies during submarine missions. AB - Relations between personality profiles, measured by the Personality Characteristics Inventory (PCI), and habitual coping strategies, measured by the Utrecht Coping List (UCL), were investigated in a sample of submarine personnel and office employees. The predictive validity of these instruments were examined for reported stress, health complaints, and salivary cortisone measures during 3 submarine missions. PCI and UCL were completed before the missions, and questionnaires and saliva were collected weekly. The results showed no significant relations between PCI profiles and coping strategies. Interpersonal orientation, achievement motivation, and habitual coping strategies were predictors for coping during the submarine missions. Problem-directed strategies and interpersonal sensitivity combined with strong achievement motivation were related to low indicated stress from social factors (lack of privacy, interpersonal tension, and crowding) and homesickness. The findings suggest that interpersonal characteristics need to be considered in the selection of submariners and personnel for other military settings in which units are exposed to prolonged stress and isolation. PMID- 14552348 TI - Psychological issues relevant to astronaut selection for long-duration space flight: a review of the literature. AB - This technical paper reviews the current literature on psychological issues relevant to astronaut selection for long-duration space flights. Interpersonal problems have been and remain a recurring problem for both short and long duration space flights. Even after completion of the space mission, intense psychological aftereffects are reported. The specific behavioral problems experienced during United States and Soviet Union space flights are reviewed, specifically addressing contentious episodes and impaired judgments that occurred during the Mercury, Apollo, and Skylab missions. Psychological tests used in the selection process for the space program have focused primarily on the detection of gross psychopathologies in potential candidates. Although these psychological instruments excluded some people from becoming astronauts, the battery of tests failed to predict which individuals would manifest behavioral aberrations in judgment, cooperative functioning, overt irritability, or destructive interpersonal actions. PMID- 14552349 TI - How we will go to Mars. AB - This article studies the efficiency of ejecting waste generated by the life support system (LSS) of a manned spacecraft to reduce initial mass on low earth orbit. The spacecraft is used for a long-duration interplanetary mission and is equipped with either a chemical or a nuclear-thermal propulsion system. For this study we simulate an optimal control problem for a given spacecraft maneuver. An impulsive approximation of the optimal interplanetary spacecraft trajectory is assumed, which allows us to reduce the general optimal control problem to hierarchic structure of 'outer' and 'inner' subproblems. This structure is analyzed using the Pontryagin's Maximum principle. Numerical results, illustrating the efficiency of waste ejection are shown for typical Earth-Mars transfer trajectories. This results confirm in theory that using a waste ejection system makes an early manned Mars mission possible without having to design and build new, advanced biological LSS. PMID- 14552350 TI - Temporal changes in the bacterioplankton of a Northeast Ohio (USA) River. AB - To examine temporal changes in a bacterial community, water samples were collected monthly for one year from five sites along a major use-reuse river, the Cuyahoga River, in northeastern Ohio (USA). Fluorescent in situ hybridization (FISH) was used to enumerate population sizes of two species of common bacteria, Pseudomonas putida and Acinetobacter calcoaceticus; FISH was also performed with a Domain Bacteria specific probe. In addition, the total bacteria (based on DAPI staining), colony forming units (on modified Nutrient agar) and coliforms were enumerated and supporting physical/chemical data were collected. Each variable examined exhibited a different seasonal pattern. Temporal changes in total number of bacteria and population size of P. putida were correlated with turbidity and precipitation suggesting that allochthonous sources and scouring of the benthos may be major contributors to these portions of the community. In contrast, the number of cells hybridizing the Domain Bacteria and A. calcoaceticus probes were correlated with temperature. Thus, different aspects of the bacterial community are potentially controlled by different factors and the role of allochthonous and autochthonous sources may vary among species. PMID- 14552351 TI - Hyper-gravity effects on the Arabidopsis transcriptome. AB - Callus cultures of Arabidopsis thaliana (cv. Columbia) in Petri dishes were exposed to altered g-forces by centrifugation (1-10 g). Using semi-quantitative RT-PCR transcripts of genes coding for metabolic key enzymes (ADP-glucose pyrophosphorylase, ADPG-PP; beta-amylase, fructose-1,6-bisphosphatase, FBPase; glyceraldehyde-P dehydrogenase, GAPDH; hydroxymethylglutaryl-CoA reductase, HMG; phenylalanine-ammonium-lyase, PAL; PEP carboxylase, PEPC) were used to monitor threshold conditions for g-number (all) and time of exposure (beta-amylase) which led to altered amounts of the gene product. Exposure to approximately 5 g and higher for 1 h resulted in altered transcript levels: transcripts of beta amylase, PAL, and PEPC were increased, those of ADPG-PP decreased, while those of FBPase, GAPDH, and HMG were not affected. This probably indicates a shift from starch synthesis to starch degradation and increased rates of anaplerosis (PEPC: supply of ketoacids for amino acid synthesis). In order to get more information about g-related effects on gene expression, we used a 1-h exposure to 7 g for a microarray analysis, using a commercial A. thaliana chip with 4105 unique annotated clusters/genes (IncyteGenomics). Transcripts of more than 200 genes were significantly increased in amount (ratio 7 g/1 g control; 2(1.6) and larger). They fall into several categories. Transcripts coding for enzymes of major pathways form the largest group (25%), followed by gene products involved in cellular organization and cell wall formation/rearrangement (17%), signalling, phosphorylation/dephosphorylation (12%), proteolysis and transport (10% each), hormone synthesis plus related events (8%), defense (4%), stress-response (2%), and gravi-sensing (2%). Many of the alterations are part of a general stress response, but some changes related to the synthesis/rearrangement of cell wall components could be more hyper-g-specific. We only found few gene products, which were decreased in relation to 1 g controls, and these were less significant (ratio < 2(1.6)). We thus assume that g-forces above a threshold of about 5 g for 1 h are sensed by plant cells in general, causing distinct metabolic responses, which obviously in part, are regulated by gene expression. PMID- 14552352 TI - Phylogeny and systematic position of Opiliones: a combined analysis of chelicerate relationships using morphological and molecular data. AB - The ordinal level phylogeny of the Arachnida and the suprafamilial level phylogeny of the Opiliones were studied on the basis of a combined analysis of 253 morphological characters, the complete sequence of the 18S rRNA gene, and the D3 region of the 28S rRNA gene. Molecular data were collected for 63 terminal taxa. Morphological data were collected for 35 exemplar taxa of Opiliones, but groundplans were applied to some of the remaining chelicerate groups. Six extinct terminals, including Paleozoic scorpions, are scored for morphological characters. The data were analyzed using strict parsimony for the morphological data matrix and via direct optimization for the molecular and combined data matrices. A sensitivity analysis of 15 parameter sets was undertaken, and character congruence was used as the optimality criterion to choose among competing hypotheses. The results obtained are unstable for the high-level chelicerate relationships (except for Tetrapulmonata, Pedipalpi, and Camarostomata), and the sister group of the Opiliones is not clearly established, although the monophyly of Dromopoda is supported under many parameter sets. However, the internal phylogeny of the Opiliones is robust to parameter choice and allows the discarding of previous hypotheses of opilionid phylogeny such as the "Cyphopalpatores" or "Palpatores." The topology obtained is congruent with the previous hypothesis of "Palpatores" paraphyly as follows: (Cyphophthalmi (Eupnoi (Dyspnoi + Laniatores))). Resolution within the Eupnoi, Dyspnoi, and Laniatores (the latter two united as Dyspnolaniatores nov.) is also stable to the superfamily level, permitting a new classification system for the Opiliones. PMID- 14552353 TI - Adaptation of SUBSTOR for controlled-environment potato production with elevated carbon dioxide. AB - The SUBSTOR crop growth model was adapted for controlled-environment hydroponic production of potato (Solanum tuberosum L. cv. Norland) under elevated atmospheric carbon dioxide concentration. Adaptations included adjustment of input files to account for cultural differences between the field and controlled environments, calibration of genetic coefficients, and adjustment of crop parameters including radiation use efficiency. Source code modifications were also performed to account for the absorption of light reflected from the surface below the crop canopy, an increased leaf senescence rate, a carbon (mass) balance to the model, and to modify the response of crop growth rate to elevated atmospheric carbon dioxide concentration. Adaptations were primarily based on growth and phenological data obtained from growth chamber experiments at Rutgers University (New Brunswick, N.J.) and from the modeling literature. Modified SUBSTOR predictions were compared with data from Kennedy Space Center's Biomass Production Chamber for verification. Results show that, with further development, modified-SUBSTOR will be a useful tool for analysis and optimization of potato growth in controlled environments. PMID- 14552354 TI - Classification and comparison of biomechanical response strategies for accommodating landing impact. AB - The purposes of this study were to (a) present a theoretical model to explain the methods by which individuals accommodate impact force in response to increases in an applied stressor, (b) use the model and a correlation procedure to classify a sample of individuals based on their observed response patterns, and (c) statistically evaluate the classification process. Ten participants performed landings from three heights while video and force platform data were being collected. Magnitudes of impact-force characteristics from ground reaction force and lower extremity joint moments were evaluated relative to changes in landing momentum. Correlation between impact force and landing momentum was used to classify participant responses into either a positive or negative biomechanical strategy, as defined by the model. Positive and negative groups were compared using the Mann-Whitney U test. Results indicated that all responses fit within the categories defined by the model. Some individuals preferred positive strategies while others preferred negative ones depending on the specific variable. Only one participant consistently exhibited the negative strategy for all variables. Positive and negative groups were determined to be statistically different, p < or = 0.05, for 61% of the comparisons, suggesting actual differences between groups. The proposed model appeared robust and accounted for all responses in the current experiment. The model should be evaluated further using landing and other impact activities; it should be refined and used to help researchers understand individual impact-response strategies in order to identify those who may be at risk for impact related injuries. PMID- 14552355 TI - The physical price of a ticket into space. AB - As a direct consequence of exposure to microgravity astronauts experience a number of physiological changes, which can have serious medical implications when they return to Earth. Most immediate and significant are the head-ward shift of body fluids and the removal of gravitational loading from bone and muscles, which lead to progressive changes in the cardiovascular and musculoskeletal systems. Cardiovascular adaptations result in an increased incidence of orthostatic intolerance (fainting) post-flight, decreased cardiac output and reduced exercise capacity. Changes in the musculoskeletal system contribute significantly to the impaired functions experienced in the post-flight period. The underlying factor producing these changes is the absence of gravity. Countermeasures, therefore, are designed primarily to simulate Earth-like movements, stresses and system interactions. Exercise is one approach that has received wide operational use and acceptance in both the US and Russian space programmes, and has enabled humans to stay relatively healthy in space for well over a year. Although it remains the most effective countermeasure currently available, significant physiological degradation still occurs. The development of other countermeasures will therefore be necessary for longer duration missions, such as the human exploration of Mars. PMID- 14552356 TI - Theory for a gas composition sensor based on acoustic properties. AB - Sound travelling through a gas propagates at different speeds and its intensity attenuates to different degrees depending upon the composition of the gas. Theoretically, a real-time gaseous composition sensor could be based on measuring the sound speed and the acoustic attenuation. To this end, the speed of sound was modelled using standard relations, and the acoustic attenuation was modelled using the theory for vibrational relaxation of gas molecules. The concept for a gas composition sensor is demonstrated theoretically for nitrogen-methane-water and hydrogen-oxygen-water mixtures. For a three-component gas mixture, the measured sound speed and acoustic attenuation each define separate lines in the composition plane of two of the gases. The intersection of the two lines defines the gas composition. It should also be possible to use the concept for mixtures of more than three components, if the nature of the gas composition is known to some extent. PMID- 14552357 TI - Describing functional requirements for knowledge sharing communities. AB - Human collaboration in distributed knowledge sharing groups depends on the functionality of information and communication technologies (ICT) to support performance. Since many of these dynamic environments are constrained by time limits, knowledge must be shared efficiently by adapting the level of information detail to the specific situation. This paper focuses on the process of knowledge and context sharing with and without mediation by ICT, as well as issues to be resolved when determining appropriate ICT channels. Both technology-rich and non technology examples are discussed. PMID- 14552358 TI - Anaerobic conditions improve germination of a gibberellic acid deficient rice. AB - Dwarf plants are useful in research because multiple plants can be grown in a small area. Rice (Oryza sativa L.) is especially important since its relatively simple genome has recently been sequenced. We are characterizing a gibberellic acid (GA) mutant of rice (japonica cv 'Shiokari,' line N-71) that is extremely dwarf (20 cm tall). Unfortunately, this GA mutation is associated with poor germination (70%) under aerobic conditions. Neither exogenous GA nor a dormancy breaking heat treatment improved germination. However, 95% germination was achieved by germinating the seeds anaerobically, either in a pure N2 environment or submerged in unstirred tap water. The anaerobic conditions appear to break a mild post-harvest dormancy in this rice cultivar. PMID- 14552359 TI - Sensitivity of wheat and rice to low levels of atmospheric ethylene. AB - Ethylene (C2H4) gas is produced throughout the life cycle of plants and can accumulate in closed growth chambers to levels 100 times higher than in outside environments. Elevated atmospheric C2H4 can cause a variety of abnormal responses, but the sensitivity to elevated C2H4 is not well characterized. We evaluated the C2H4 sensitivity of wheat (Triticum aestivum L.) and rice (Oryza sativa L.) in five studies. The first three studies compared the effects of continuous C2H4 levels ranging from 0 to 1000 nmol mol-1 (ppb) in a growth chamber throughout the life cycle of the plants. A short-term 1000 nmol mol-1 treatment was included in which exposure was stopped at anthesis. Yield was reduced by 36% in wheat and 63% in rice at 50 nmol mol-1 and both species were virtually sterile when continuously exposed to 1000 nmol mol-1. However, the yield reductions were much less with exposure that stopped at anthesis, suggesting the detrimental effect of C2H4 on yield was greatest around the time of seed set. Two additional studies evaluated the differential sensitivity of two wheat cultivars (Super Dwarf and USU-Apogee) to 50 nmol mol-1 C2H4 at three CO2 levels [350, 1200, 5000 micromoles mol-1 (ppm)] in a greenhouse. Yield of USU Apogee was not significantly reduced by C2H4 but the yield of Super Dwarf was reduced by 60%. Elevated CO2 did not influence the sensitivity to C2H4. A difference in the C2H4 sensitivity of USU-Apogee between greenhouse and growth chamber trials suggests that C2H4 sensitivity is dependent on the environment. Collectively, the data suggest that relatively low levels of C2H4 could induce anomalous plant responses by accumulation in greenhouses and growth chambers with inadequate ventilation. The data also suggest that C2H4 sensitivity can be reduced by both genetic and environmental manipulations. PMID- 14552360 TI - Protein nano-crystallogenesis. AB - We demonstrate the feasibility of growing crystals of protein in volumes as small as 1 nanoliter. Advances in the handling of very small volumes (i.e. through inkjet and other technologies) open the way towards fully automated systems. The rationale for these experiments is the desire to develop a system that speeds up the structure determination of proteins by crystallographic techniques, where most of the precious protein sample is wasted for the identification of the ideal crystallisation conditions. An additional potential benefit of crystallisation in very small volumes is the potential improvement of the crystal quality through reduced convection during crystal growth. Furthermore, in such small volumes even very highly supersaturated conditions can be stable for prolonged periods, allowing additional regions of phase-space to be prospected for elusive crystallisation conditions. A massive improvement in the efficiency of protein crystallogenesis will cause a paradigm shift in the biomolecular sciences and will have a major impact in product development in (for example) the pharmaceutical industry. PMID- 14552361 TI - [Secondary adrenal gland insufficiency]. PMID- 14552362 TI - Influence of dietary urea level on digestive function and growth performance of cattle fed steam-flaked barley-based finishing diets. AB - Four Holstein steers (282 kg) with cannulas in the rumen and proximal duodenum were used in a 4 x 4 Latin square experiment to evaluate the influence of dietary urea level (0, 0.4, 0.8, and 1.2%, DM basis) in a steam-flaked barley-based finishing diet on digestive function. There were no treatment effects (P > 0.20) on ruminal digestion of OM and ADF. Increasing dietary urea level increased (linear, P < 0.01) ruminal starch digestion. Ruminal degradability of protein in the basal diet (no supplemental urea) was 60%. Increasing dietary urea level did not increase (P > 0.20) ruminal microbial protein synthesis or nonammonia N flow to the small intestine. There were no treatment effects (P > 0.20) on total-tract ADF digestion. Total tract digestion of OM (quadratic, P < 0.01) and starch (linear, P < 0.05) increased slightly with increasing urea level. Urea supplementation increased (linear, P < 0.01) ruminal pH 1 h after feeding; however, by 3 h after feeding, ruminal pH was lower (cubic, P < 0.05) with urea supplemented diets. Urea supplementation did not affect (P > 0.20) ruminal molar proportions of acetate and propionate. One hundred twenty crossbred steers (252 kg; approximately 25% Brahman breeding) were used in an 84-d feeding trial (five pens per treatment) to evaluate treatment effects on growth performance. Daily weight gain increased (linear, P = 0.01) with increasing urea level, tending to be maximal (1.53 kg/d; quadratic, P = 0.13) at the 0.8% level of urea supplementation. Improvements in ADG were due to treatment effects (linear, P < 0.01) on DMI. Urea supplementation did not affect (P > 0.20) the NE value of the diet for maintenance and gain. Observed dietary NE values, based on growth performance, were in close agreement with expected based on tabular values for individual feed ingredients, averaging 100.4%. We conclude that with steam-flaked barely-based finishing diets, ruminal and total-tract digestion of OM and ruminal microbial protein synthesis may not be increased by urea supplementation. In contrast, ADG was optimized by dietary inclusion of 0.8% urea. Urea supplementation may not enhance the net energy value of steam-flaked barely-based finishing diets when degradable intake protein is greater than 85% of microbial protein synthesis. PMID- 14552363 TI - Body condition at parturition and postpartum weight changes do not influence the incidence of short-lived corpora lutea in postpartum beef cows. AB - Seventy-seven multiparous beef cows (Hereford and Angus x Hereford) with thin to moderate BCS at calving were used to evaluate the effects of body condition at parturition and BW change after calving on duration and occurence of luteal activity before and after first estrus. Blood samples were collected twice weekly after parturition to determine the occurrence of the first postpartum luteal activity (LA, progesterone > or = 0.5 ng/mL). Weight changes and BCS were determined at 2-wk intervals. Cows were exposed to bulls and observed twice daily for behavioral estrus. Luteal activity was classified as normal if plasma concentrations of progesterone were > or = 0.5 ng/mL for at least 11 d, or short if concentrations of progesterone were > or = 0.5 ng/mL for 10 d or less. The interval from parturition to first normal LA was shorter (P < 0.001) for moderate condition (BCS > or = 4.5) than for thin (BCS < or = 4) cows (58.3 +/- 3.2 vs. 93.3 +/- 5.1 d, respectively). Interval to first estrus also was shorter (P < 0.001) for moderate than for thin cows (53.3 +/- 3.7 vs. 89.3 +/- 5.6 d, respectively). Before the first normal LA, 78% of cows had an increase in progesterone for < 11 d. Postpartum weight change and BCS at calving did not influence the incidence of estrus associated with first normal LA. After the first estrus, 72% of cows had normal LA, 16% had a short luteal phase, and 12% lacked LA. Postpartum weight change and BCS did not influence the length of LA associated with the first estrus. Cows with normal LA had increased (P < 0.05) maximal concentrations of progesterone compared with cows that had a short luteal phase. When a transient increase in progesterone occurred before first behavioral estrus, 81% of cows had normal luteal function after estrus. We conclude that when beef cows are in thin to moderate body condition at calving, postpartum BW change and BCS at calving do not influence the duration of luteal activity before or after the first postpartum estrus. PMID- 14552364 TI - Effect of repeated administration of combination trenbolone acetate and estradiol implants on growth, carcass traits, and beef quality of long-fed Holstein steers. AB - Our objective was to determine the effect of repeated use of implants on feedlot performance and carcass characteristics of Holstein cattle. Holstein steers (n = 128) weighing an average of 211 kg were blocked by weight and randomly assigned to 16 pens. At the start of the trial (d 0), pens were assigned to one of four treatments: 1) nonimplanted control (C); 2) implant on d 0, 112, and 224 (T3); 3) implant on d 112 and 224 (T2); and 4) implant on d 224 (T1). Component TE-S implants (120 mg of trenbolone acetate and 24 mg of estradiol per implant) were used for all treatments during the 291-d feeding period. Over the course of the study, T2 and T3 cattle had greater ADG and final weights than C and T1 cattle (P < 0.05). Steers were harvested at a commercial abattoir on d 291. Hot carcass weights of T3 steers were greater than those of C and T1 steers (P < 0.05). Dressing percentage, adjusted 12th-rib fat, percentage of kidney, pelvic, and heart fat, yield grade, and longissimus color were not different among treatments (P > or = 0.26). Longissimus muscle areas (LMA) of T2 and T3 carcasses were larger than LMA of C (P < 0.01). No USDA Select carcasses were produced from C cattle, whereas the percentage of Select carcasses from implanted cattle ranged from 10 to 18%. Skeletal maturity advanced (P < 0.05) progressively with each additional implant. Steaks from T3 carcasses had a higher percentage of protein than controls (P < 0.05) and were less tender than all other treatments (P < 0.05). Repeated administration of combination trenbolone acetate and estradiol implants increased ADG and resulted in heavier carcasses with larger LMA. Administration of three successive implants decreased tenderness of Holstein beef, and resulted in more advanced skeletal maturity scores. PMID- 14552365 TI - Mortality of Creole kids during infection with gastrointestinal strongyles: a survival analysis. AB - Mortality due to strongyles infection in small ruminants is a critical component of flock productivity in a tropical climate. In goat production, few experiments have been conducted to estimate the variability of this trait. A survival analysis study was carried out in the Creole experimental flock of INRA-Gardel (Moule, Guadeloupe) to identify management and genetic factors influencing mortality of kids reared at pasture and infected with gastrointestinal strongyles, predominantly Haemonchus contortus and Trichostrongylus colubriformis. Survival curves from 3 and 11 mo of age were analyzed for 837 kids sired by 48 bucks and 250 does. The causes of death were recorded. Mortality due to gastrointestinal strongyles was the variable considered. The flock management included drenchings with levamisole every 8 wk. Fecal egg counts and packed cell volume were regularly measured after 7 wk of natural infection. All but 6.7% of the records were uncensored, with an average failure time of 165 d. The probability of death following gastrointestinal infection was more than three times greater in males than in females. Kids raised by their mother before weaning had a lower (P < 0.05) relative risk of dying than those reared in nursery (0.40 vs. 1). Parity of the dam and litter size effects were not significant. The risk of death was reduced by approximately 80% during the 3 wk that followed a drenching (P < 0.01). Risk decreased by about 25% for each additional kilogram of body weight at weaning. Live weight, fecal egg counts, and packed cell volume all had significant effects on risk of death when introduced as time-dependent covariates in the model (P < 0.0001 for live weight and packed cell volume, and P < 0.01 for fecal egg counts). The estimated genetic variability was small and inaccurate. These results demonstrated that risk of death from gastrointestinal infection could be reduced with appropriate flock management. High infection levels increased the risk of death, but they were not the direct cause. The important mediation of reduced body weight and anemia in likelihood of death is highlighted. More data are needed to better assess the possibility for genetic improvement of viability in Creole kids during gastrointestinal strongyle infection. PMID- 14552366 TI - Design and standards for genetic evaluation of swine seedstock populations. AB - The purpose of this article is to describe a program for evaluation of seedstock populations in the swine industry. Differences among seedstock populations for economically important traits must be identified in order for pork producers to efficiently use available genetic resources. National genetic evaluation programs have the potential to identify the important differences among populations and to increase the rate of genetic improvement in a population. Program results provide performance benchmarks that stimulate testing and selection procedures by seedstock suppliers that further increase the rate of genetic improvement. A Terminal Sire Line Genetic Evaluation Program was designed and conducted in the United States by the National Pork Producers Council (Des Moines, IA) to compare seedstock populations for use in crossbreeding systems. High levels of statistical accuracy for program results were established; the ability to detect differences of 0.25 SD per trait, a power of test of 75%, and a 5% significance level were selected. Pure breeds and breeding company sire lines were nominated for the program. Semen was collected from nominated boars and distributed to cooperating commercial producers during eight 1-wk breeding periods. Pigs were produced in 136 commercial herds and transported to testing facilities at 8 to 23 d of age. Nine of the 11 sire lines originally entered in the program completed the sampling requirements for statistical analysis. High levels of statistical accuracy and a large, representative sample of boars with restrictions on genetic relationships ensured that the program results included unbiased, highly accurate sire line data for growth, carcass, meat quality, and eating quality traits of economic importance. This program has shown commercial producers that they have several choices of sire lines for changing their crossbreeding programs in desired trait areas. Commercial product evaluation must be an ongoing process, and this program serves as a model for future testing and evaluation of diverse genetic seedstock populations. PMID- 14552367 TI - Stillbirth in the pig in relation to genetic merit for farrowing survival. AB - The objectives of this study were to analyze the incidence of different categories of stillborn piglets in relation to genetic merit for farrowing survival of sows and litters and to analyze relationships of total number of piglets born per litter, average BW of the litter, and within-litter variation in BW with genetic merit for farrowing survival of sows and litters. Records of 336 purebred litters, produced by 307 first-to eighth-parity sows, were collected on a nucleus farm in Brouennes, France. Breeding values for farrowing survival were estimated for sows (EBVfs_maternal) and litters (EBVfs_direct) using a large data set from which information obtained in the current study was excluded. For each litter, BW, number of stillborn piglets (classified as nonfresh stillborn, prepartum stillborn, intrapartum stillborn, and postpartum stillborn), and number of live-born piglets were recorded. Birth weights of stillborn piglets were lower than BW of live-born piglets (P < 0.0001), except for prepartum stillbirths. The total number of stillborn piglets per litter and the number of stillborn piglets in each category decreased with increasing EBVfs_maternal (P < 0.01). An increase in EBVfs_direct was also associated with a decrease in the total number of stillborn piglets per litter (P < 0.01). This decrease was due to a decrease in the number of nonfresh, prepartum, and postpartum stillborn piglets but not to a decrease in the number of intrapartum stillborn piglets. Probabilities of stillbirth in relation to EBVfs_maternal were higher than probabilities of stillbirth in relation to EBVfs_direct. Total number of piglets born decreased with increasing EBVfs_direct (P = 0.0003), but was not related to EBVfs_maternal. Average BW of the litter (P < 0.0001) and within-litter variation in BW (P = 0.05) decreased with increasing EBVfs_maternal but were not related to EBVfs_direct. Selection for the maternal genetic component of farrowing survival seems a better strategy than selection for the direct genetic component. Selection for the maternal genetic component of farrowing survival reduces stillbirth in all categories and does not affect litter size. PMID- 14552368 TI - Genetic evaluation of an index of birth weight and yearling weight to improve efficiency of beef production. AB - The CGC population is a stabilized composite of 1/2 Red Angus, 1/4 Charolais, and 1/4 Tarentaise germplasm. The objectives of this research were to estimate genetic parameters for weight traits of CGC and to evaluate genetic responses resulting from selection based on the following index: I = 365-d weight 3.2(birth weight). Phenotypes evaluated were birth weight (n = 5,083), 200-d weight (n = 4,902), 365-d weight (n = 4,626), and the index. In addition, there were 1,433 cows with at least one recorded weight, and 4,375 total observations of cow weight collected at the time their calves were weaned. In 1989, a randomly selected control line and a line selected for greater values of the index were established. Average generation intervals were 3.16 +/- 0.04 and 3.90 +/- 0.08 yr in the index and control lines, respectively. The index selection line (n = 950) accumulated approximately 212 kg more selection differential than the control line over three generations (n = 912). Heritability estimates for direct effects were 0.32 +/- 0.04, 0.49 +/- 0.05, 0.49 +/- 0.05, 0.30 +/- 0.04, and 0.70 +/- 0.04 for the index, birth weight, 365-d weight, 200-d weight, and cow weight, respectively. Heritability estimates for maternal effects were 0.05 +/- 0.02, 0.11 +/- 0.03, 0.04 +/- 0.02, and 0.19 +/- 0.04 for the index, birth weight, 365 d weight, and 200-d weight, respectively. In the control line, direct genetic changes for the index and its components were small. For the index selection line, direct genetic changes for the index, birth weight, 365-d weight, 200-d weight, and cow weight were 6.0 +/- 0.3, 0.45 +/- 0.09, 7.74 +/- 0.55, 3.42 +/- 0.25, and 6.3 +/- 0.9 kg/generation, respectively. Maternal genetic changes were generally small for both the control and index selection lines. Thus, selection for the index produced positive correlated responses for direct genetic effects on BW traits at all ages, with only minor effects on maternal genetic effects. Results demonstrate that despite a genetic antagonism that compromises selection response for decreased birth weight and increased postnatal growth, favorable genetic responses can be achieved with the selection index used in this study. PMID- 14552369 TI - Influence of litter size and creep feeding on preweaning gain and influence of preweaning growth on growth to slaughter in barrows. AB - The importance of birth-to-weaning average daily gain as a determinant of weight at a final age and yield of marketable pork was investigated. Treatments were imposed to create variation in birth-to-weaning ADG independent of birth weight. Newborn pigs were cross-fostered to create litters of four through 14 pigs/litter. Creep feed was offered to pigs from 5 d of age or during last 2 d before weaning at 13 to 20 d (average = 17 d). Growth rate and carcass dissection data were obtained from 195 barrows that were slaughtered at an average age of 170 d (SD = 7.5), weight of 109 kg (SD = 10.5). All traits measured were influenced by birth dam and sire (P < 0.01). Quadratic and cubic effects (P < 0.09) of litter size on birth-to-weaning ADG and weaning weight were different between the creep feeding treatments. Data revealed a positive influence (P < 0.04) of creep feeding from 5 d of age on birth-to-weaning ADG and weaning weight in larger size (> 8) litters. Importance of the independent variables birth weight, birth-to-weaning ADG, weaning weight, and birth weight plus birth-to weaning ADG in determination of measures of postweaning growth and yield of marketable pork were examined by step-down regression analysis. Initial models included the linear and quadratic effects of the independent variables. In general, R2 for models ranked birth weight < birth-to-weaning ADG < d-17 weaning weight < birth weight + birth-to-weaning ADG. The R2 of models for BW at 170 d of age were 0.11 (P < 0.01) using birth weight as the independent variable, 0.16 (P < 0.01) using birth-to-weaning ADG, 0.19 (P < 0.01) using d-17 weaning weight, and 0.21 (P < 0.01) using birth weight + birth-to-weaning ADG. The model for effect of birth-to-weaning ADG on BW at 170 d of age indicated that a 10-g advantage in birth-to-weaning ADG produced a 0.94-kg advantage in BW at 170 d of age. Positive relationships (P < 0.05) between birth-to-weaning ADG and measures of postweaning growth and carcass yield suggest management practices that increase birth-to-weaning ADG may be advantageous in pork production. PMID- 14552370 TI - Effect of sire on mu- and m-calpain activity and rate of tenderization as indicated by myofibril fragmentation indices of steaks from Brahman cattle. AB - The objectives of this study were to assess the influence of sire on mu- and m calpain activities, to evaluate the relationships of activities of these enzymes to other traits related to beef palatability, and to assess the influence of sire on the rate of tenderization (as measured by myofibril fragmentation index [MFI]) in Brahman longissimus muscle. Brahman calves (n = 87), sired by nine bulls, were born, weaned, fed, and slaughtered in central Florida. Traits evaluated were mu- and m-calpain activities and MFI after 1, 7, 14, and 21 d of aging. Other traits were analyzed to determine their associations with mu- and m-calpain activity and MFI, including calpastatin activity, percentage of raw and cooked lipids, Warner Bratzler shear force (WBSF) values after 7, 14, and 21 d of aging, and sensory panel rating of tenderness, juiciness, and connective tissue amount after 14 d of aging. Data were analyzed using a model with sire, sex, year, and slaughter group (calves of the same sex slaughtered on the same date) as fixed effects, and adjusted to a constant adjusted 12th-rib fat thickness. Sire affected mu-calpain activity (P < 0.04), calpastatin activity (P < 0.01), d-14 MFI (P < 0.02), d-7 WBSF (P < 0.05), d-14 WBSF (P < 0.04), and sensory panel juiciness score (P < 0.01), but not (P < 0.75) m-calpain activity. Measures of tenderness and palatability were generally moderately to strongly correlated (both simple and residual correlations) with calpastatin and m-calpain activity. Myofibril fragmentation index residuals (adjusted for all model components except sire) after all aging periods were fitted using nonlinear regression to the exponential curve (MFI(i) = kappa0 + kappa1 exp[kappa2 t(i)] + epsilon(i), where t(i) represents aging in days, k0 is ultimate MFI after aging, kappa1 is the difference between initial and ultimate MFI, kappa2 is the rate of increase in MFI, and epsilon(i) is the error term associated with the ith observation, assumed to be independent and identically distributed normally). Sires had different estimates and combinations of estimates, which were used to plot MFI change with time. These curves visually differed for sires and suggested that postmortem tenderization extent and rate differ as well. Use of a combination of these estimated parameters in a selection/carcass sorting program represents an alternative consideration for tenderization improvement programs. PMID- 14552371 TI - Effect of age at slaughter on carcass traits and meat quality of Italian heavy pigs. AB - Barrows and gilts (n = 128) from four breed crosses were used to investigate the effect of age at slaughter on carcass traits, proteolytic enzyme activity, and meat and fat quality. Pigs were blocked by breed cross into four blocks, and within blocks, one pen (eight barrows and eight gilts) was assigned randomly to be slaughtered at either 8 or 10 mo of age. Pigs were fed a corn-barley-soybean meal finisher diet from 104 +/- 2.5 d of age (37.7 +/- 0.33 kg BW) to the appropriate slaughter age. Carcasses from older (10 mo) pigs had lower (P < 0.01) muscularity indexes and lean cut yields than those of younger (8 mo) pigs, but dressing percentage and longissimus muscle area increased (P < 0.01) with age. Older pigs produced a redder (P < 0.01) and darker (P < 0.05) semimembranosus, with lower (P < 0.01) ultimate pH and cathepsin B and B + L activities, as well as higher (P < 0.01) aminopeptidase hydrolyzing activity than younger pigs. Moreover, the longissimus muscle of pigs slaughtered at 10 mo of age had lower (P < 0.01) drip and cooking loss percentages than that from pigs slaughtered at 8 mo of age. Ham subcutaneous fat from 10-mo-old pigs had greater (P < 0.05) percentages of oleic acid and lower (P < 0.01) proportions of moisture, linoleic, and linolenic acids than subcutaneous fat from pigs slaughtered at 8 mo of age. Results from this study indicate that fresh hams from pigs slaughtered at 10 mo of age would be more suitable for the production of high-quality, Italian, dry cured hams. PMID- 14552372 TI - Carcass, sensory, and adipose tissue traits of Brangus steers fed casein formaldehyde-protected starch and/or canola lipid. AB - We predicted that providing rumen-protected starch to the small intestine would increase adiposity of intramuscular adipose tissue, and hence marbling scores. Eighteen 15-mo-old Brangus steers were assigned randomly to one of three dietary treatment groups: 1) cracked corn (Corn); 2) casein-formaldehyde-protected lipid (Canola Lipid); or 3) casein-formaldehyde-protected starch (Marble Plus). All diets were equally balanced for ME (2.91 Mcal/kg), CP (12.5%), and DM (89%). Ether extract was 3.7, 6.9, and 6.9% for the Corn, Canola Lipid, and Marble Plus diets, respectively, and the Marble Plus also contained 3.7% protected starch. Steers were fed the diets for 126 d before slaughter. Average daily feed intake (as-fed basis), ADG, and feed:gain ratio (P > or = 0.23) did not differ among treatments. Carcasses across treatments did not differ (P = 0.26) in adjusted fat thickness, longissimus muscle area, hot carcass weight, dressing percentage, marbling scores, or USDA quality grade. Percentage of kidney, pelvic, and heart fat was higher (P < 0.01) and USDA yield grade tended (P = 0.08) to be higher, for carcasses from Canola Lipid- and Marble Plus-fed steers than for carcasses from Corn-fed steers. Of the descriptive meat sensory attributes, connective tissue amount (P = 0.06) and painty flavor (P = 0.12) tended to be greater in meat from Marble Plus steers than from Canola Lipid steers. Percentages of 18:2n 6 and 18:3n-3 were higher (P < 0.01), and 15:0, 16:0, and 17:0 were lower (P < or = 0.07) in tissues from Canola Lipid- and Marble Plus-fed steers than in Corn-fed steers. Mean adipocyte volume was greater (P = 0.02) in i.m. adipose tissue and tended (P = 0.11) to be greater in s.c. adipose tissue of Canola Lipid steers (848 pL) vs. Corn steers (536 pL). Glucose incorporation into total lipids, glyceride-glycerol, and fatty acid fractions was highest (P < 0.01) in s.c. adipose tissue from steers fed Marble Plus but was unaffected (P > or = 0.33) by diet in i.m. adipose tissue. Fatty acid synthetase activity tended (P = 0.08) to be higher in s.c. adipose tissue of Marble Plus steers, and NADP-malic dehydrogenase activity was higher (P = 0.03) in i.m. adipose tissue of Canola Lipid steers. We conclude that Marble Plus did not improve carcass quality, but also did not reduce beef sensory attributes. Any differences we observed in carcass characteristics, adipose tissue cellularity, or lipogenesis apparently were caused by the protected lipid rather than the protected starch. PMID- 14552373 TI - Creatine monohydrate supplemented in swine finishing diets and fresh pork quality: III. Evaluating the cumulative effect of creatine monohydrate and alpha lipoic acid. AB - The objective of this study was to evaluate short-duration supplementation of alpha-lipoic acid (ALA) and creatine monohydrate (CMH) to improve fresh pork quality. Forty-eight commercial hybrid barrows were blocked by BW and randomly allotted to one of four treatments: 1) no CMH or ALA; 2) supplementation of 24 g of CMH(-1) x pig(-1) x d(-1); 3) supplementation of 600 mg ALA(-1) x pig(-1) x d( 1); or 4) combined CMH and ALA supplements. Twelve pigs per treatment were individually penned with ad libitum access to water and a finishing diet. Treatments were hand-fed to individual pigs daily (divided into three equal doses) for 5 d before slaughter at 113 kg BW in two separate groups of 24 pigs each. Intramuscular pH was recorded at 45 min postmortem and again at 24 h in the ham semimembranosus (SM) and the longissimus muscle (LM) between the 10th and 11th rib. A Meatcheck (SFK Technology, Peosta, IA) conductivity probe was inserted in the same anatomical locations as pH measurement, providing an index value (PY) from 0 to 100 (a higher index value indicates more intact muscle cells and higher water-holding capacity). Color (L, a, b values) measurements were obtained at 24 h postmortem on the ham gluteus medius (GM), SM, and LM. Two 2.54 cm-thick loin chops were removed from the loin for determination of Warner Bratzler shear force and glycolytic potential. The intact SM and the posterior portion of the boneless loin were vacuum-packaged and stored for 7 d to determine purge loss. Creatine-supplemented pigs had a higher (P = 0.03) PY value in the SM (66.67) at 45 min postmortem than either ALA, singularly (63.50), or in the combined CMH/ALA (62.27) treatments. (A higher PY index indicates superior water holding capacity.) Lipoic acid supplementation resulted in the highest pH at 45 min (P = 0.029). These results justify further evaluation of the potential positive influence of supplementing alpha-lipoic acid to improve pork quality. PMID- 14552374 TI - Efficacy of sucrose and milk chocolate product or dried porcine solubles to increase feed intake and improve performance of lactating sows. AB - Two experiments were conducted to determine the voluntary feed intake and performance of lactating sows fed diets containing a sucrose/milk chocolate product (MCP) blend (Exp. 1) or dried porcine solubles (DPS; Exp. 2). Dried porcine solubles is a coproduct of heparin extraction from porcine small intestines. In Exp. 1, mixed-parity sows (n = 108) at two research centers were assigned to a corn-soybean-meal-based diet formulated to contain 0.9% total lysine or a similar diet that contained 4% sucrose and 2% MCP on an as-fed basis. Sows were allowed ad libitum access to dietary treatments from the day of farrowing until pigs were weaned at approximately 21 d postpartum. Diet had no significant effect on voluntary feed intake of sows during lactation, backfat depth, or postweaning interval to estrus, but it had variable effects on body weight changes. Inclusion of the sucrose/MCP blend in diets elicited a 2% improvement in litter weaning weight at one research center and a 6% depression in litter weaning weight at the other center (diet x research center, P < 0.05). Litter size throughout lactation was unaffected by dietary treatment. In Exp. 2, mixed-parity sows (n = 119) at two research centers were assigned to corn-soybean meal-based diets formulated to contain 0.9% total lysine with 0, 1.5, or 3.0% added DPS. Sows were assigned to dietary treatments within research center, farrowing group, and parity at parturition. Dried porcine solubles tended to increase (P < 0.10) total feed consumed in the first 9 d of lactation and average daily feed intake over the entire lactation (6.03, 6.53, and 6.30 kg) for sows fed 0, 1.5, and 3.0% DPS, respectively. Litter size and weight on d 18 of lactation were not affected by concentration of DPS in the diet. Days from weaning to estrus and percentage of sows displaying estrus were not influenced by diet. We conclude that inclusion of the sucrose/MCP blend in the diet for lactating sows had no consistent effect on voluntary feed intake of sows and weight gain of nursing pigs. Inclusion of DPS at 1.5 or 3.0% tended to improve feed intake of lactating sows but had no significant influence on litter performance. PMID- 14552375 TI - Effect of mannan oligosaccharides on growth performance of weanling pigs. AB - Four experiments were conducted to evaluate the effects of mannan oligosaccharides (provided by Bio-Mos [BM], a product containing a minimum of 28% glucomannoprotein from S. cerevisiae) on growth performance of nursery pigs. Treatments were replicated with five to six pens of four to five pigs each. Initial BW ranged from 4.7 to 5.4 kg, and pigs were weaned at 16 to 20 d of age. Experiments 1, 2, and 4 consisted of Phase 1 (7 to 8 d), Phase 2 (12 to 14 d), and Phase 3 (7 to 8 d) periods, but Exp. 3 consisted only of Phase 1 (7 d) and 2 (14 d) periods. The diets for Phase 1, 2, and 3 contained 1.6, 1.5, and 1.1% Lys, respectively. The treatments in Exp. 1 were 0, 0.20, and 0.30% BM, which did not affect growth performance. The treatments in Exp. 2 were two levels of excess Zn (0 and 3,000 ppm) and three levels of BM (0, 0.20, and 0.30%) in a 2 x 3 factorial. Excess Zn increased (P < 0.08) ADG and ADFI in Phase 2 and 3 and overall. The 0.20% BM addition increased ADG (Phase 3 and overall) and ADFI (Phase 2 and overall) in the absence of excess Zn but did not affect or decreased these response variables in the presence of excess Zn (Zn x BM quadratic, P < 0.08). Experiment 3 was similar to Exp. 2, but the 0.30% BM addition was not used. Excess Zn decreased (P < 0.09) ADG in Phase 1 but increased (P < 0.09) ADG and ADFI in Phase 2. The BM decreased (P < 0.03) overall ADFI but increased Phase 2 and overall ADG and gain:feed (GF) in the absence of excess Zn but not in the presence of excess Zn (Zn x BM, P < 0.07). The BM decreased ADFI during Phase 2, but the decrease was greater in pigs fed excess Zn (Zn x BM, P < 0.07). Experiment 4 evaluated the interactive effects of the antibiotic (oxytetracycline and neomycin) and BM and of Zn and BM. Antibiotic (no excess Zn) increased (P < 0.01) ADG and ADFI in Phases 2 and 3 and overall. The BM addition decreased ADG and GF in Phase 2 when the antibiotic was not in the diet but increased ADG when the antibiotic was in the diet (antibiotic x BM, P < 0.05). Excess Zn increased (P < 0.07) ADG and ADFI during Phases 2 and 3 and overall. In Phase 2, the 0.20% BM decreased GF when excess Zn was not added to the diet but increased GF when Zn was included (Zn x BM, P < 0.03). Mannan oligosaccharides improved pig performance in some instances during Phase 2 when fed in combination with an antibiotic and no excess dietary Zn, but it had no effect or negative effects in the presence of excess Zn or in the absence of an antibiotic. PMID- 14552376 TI - Effect of carbohydrate source on growth performance, carcass traits, and meat quality of growing-finishing pigs. AB - Two experiments were conducted to determine the effect of substituting a more available dietary carbohydrate (CHO) for portions of corn or fat in the diet on growth performance, carcass traits, meat quality, and serum or plasma metabolites in growing-finishing pigs. A three-phase feeding program was used with corn soybean meal diets formulated to provide 105% of the Lys requirement for barrows or gilts gaining 325 g of lean daily in Exp. 1 or gilts gaining 350 g of lean daily in Exp. 2. Diets were isoenergetic within experiments. All other nutrients met or exceeded suggested requirements. In Exp. 1, pigs were allotted to three dietary treatments (0, 7.5, or 15.0% sucrose), with three replications of barrows and three replications of gilts, and with three or four pigs per replicate pen; average initial and final BW were 25.2 and 106.7 kg. In Exp. 2, gilts were allotted to two dietary treatments (waxy [high amylopectin] or nonwaxy [75% amylopectin and 25% amylose] corn as the grain source), with five replications of four gilts per replicate pen; average initial and final BW were 37.7 and 100.0 kg. In Exp. 1, ADG and gain:feed ratio increased linearly (P < 0.02) as dietary sucrose increased. Minolta color scores, a* and b*, and drip loss (P < 0.06) also increased linearly with added sucrose. In Exp. 2, ADG, carcass weight and length, and the Minolta a* value were greater for pigs fed waxy corn (P < 0.08) than for those fed nonwaxy corn. Feed intake, longissimus muscle area, 10th-rib and average backfat thickness, dressing percentage, fat-free lean, percentage of lean and muscling, lean gain per day, total fat, percentage fat, lean:fat ratio, serum or plasma metabolites (Exp. 1: serum urea N; Exp. 2: serum urea N, and plasma nonesterified fatty acids, triacylglycerols, total and high-density lipoprotein cholesterol, insulin, and total protein), pH of the longissimus muscle, and subjective muscle scores (color, firmness-wetness, and marbling) were not affected by diet in either experiment. In summary, increasing availability of dietary CHO in growing-finishing pig diets improved growth performance, but it did not affect carcass traits. PMID- 14552377 TI - Use of carbohydrases in corn-soybean meal-based nursery diets. AB - Three experiments were conducted to test the hypothesis that supplementing nursery pig diets with a mixture of carbohydrases (CS) will improve pig performance and nutrient digestibility. The CS used in these experiments contained 7 units/g of alpha-1,6-galactosidase, 22 units/g of beta-1,4-mannanase, beta-1,4 mannosidase, and trace amounts of other enzymes. In Exp. 1, 108 pigs weaned at d 21 of age were fed one of three diets containing 0 (control), 0.1, or 0.2% CS for 5 wk, based on a three-phase feeding program (1, 2, and 2 wk). Over the entire 35-d period, ADG was not affected (P > 0.05) by treatment, but supplementing 0.1% CS increased (P < 0.05) gain:feed by 9%. Experiment 2 used 10 gilts fitted with simple T-cannula in the terminal ileum at 3 wk of age. After cannulation, pigs were fed the same control Phase I and II diets, but the Phase III diet contained either 0 or 0.1% CS. Ileal samples were collected for the 3 d following the 5-d adjustment period during Phase III. Apparent ileal digestibility of GE, lysine, threonine, and tryptophan was greater (P < 0.05) in the CS diet. In Exp. 3, 90 pigs weaned at 21 d of age were fed the same control Phase I and II diets, but the Phase III diet contained either 0 or 0.1% CS. Phase III diets were fed for 3 wk. Average daily gain of the CS group was greater (P < 0.05) than the control group during wk 3. Gain:feed ratio was greater (P < 0.05) for the carbohydrase group during the entire Phase III period. Four pigs per treatment were killed at the end of Exp. 3 to measure villus height and to determine the concentration of raffinose and stachyose in different parts of the gastrointestinal tract. Average villus height was greater (P < 0.05) in pigs fed the CS diet. Carbohydrase supplementation decreased (P < 0.05) the concentration of stachyose in freeze-dried digesta from the proximal and distal small intestine. Raffinose concentration, on the other hand, was decreased (P < 0.05) by CS supplementation only in the distal small intestine. These lower concentrations suggest that CS improved the digestibility of carbohydrate in soybean meal. In conclusion, the addition of CS to Phase I and Phase II nursery diets containing low levels of soybean meal did not improve pig performance, but its addition to corn-soybean meal-based Phase III nursery diets improved gain:feed ratio and energy and AA digestibility. PMID- 14552379 TI - Effect of chromium propionate on growth, carcass traits, pork quality, and plasma metabolites in growing-finishing pigs. AB - Two experiments were conducted to determine the effects of dietary Cr, as Cr propionate, on growth, carcass traits, pork quality, and plasma metabolites in growing-finishing swine. Ninety-six crossbred gilts (Exp. 1; initial and final BW of 28 [SEM = 0.41] and 109 [SEM = 2.11] kg) or 144 PIC Cambrough 22 barrows (Exp. 2; initial and final BW of 26 [SEM = 0.39] and 111 [SEM = 2.52] kg) were allotted to six or four dietary treatments, respectively, with six replications and four (Exp. 1) or six (Exp. 2) pigs in each replicate pen blocked by weight in randomized complete block designs. The six dietary treatments for Exp. 1 were 1) corn-soybean meal (C-SBM), 2) C-SBM + 50 ppb Cr, 3) C-SBM + 100 ppb Cr, 4) C-SBM + 200 ppb Cr, 5) C-SBM low NE diet, and 6) C-SBM low NE diet + 200 ppb Cr. The four dietary treatments for Exp. 2 were C-SBM with 0, 100, 200, or 300 ppb Cr. Growth, carcass traits, and plasma metabolite (collected on d 29 and at each phase change) data were taken at the end of both experiments and pork quality data were taken at the end of Exp. 1. There was no effect (P > 0.10) on overall growth performance when pigs were fed graded levels of Cr (Exp. 1 and 2) or Cr in the positive control or low NE diets (Exp. 1). Longissimus muscle area, ham weight, ham fat-free lean, and total carcass lean were increased in pigs fed 200 ppb in the positive control diets but decreased in pigs fed 200 ppb Cr in the low NE diets (Cr x NE, P < 0.08). There was no effect of Cr concentration (P > 0.10) on carcass traits in Exp. 2. In Exp. 1, cook loss of a fresh or a frozen chop was decreased (P < 0.10) by 200 ppb Cr. In Exp. 1, NEFA concentration was decreased (P < 0.05) in pigs fed Cr in the positive control or low NE diets during the early-finishing period. In Exp. 2, the addition of Cr decreased NEFA (quadratic, P < 0.09) and plasma urea N (linear, P < 0.02) concentrations and tended to increase total cholesterol and high density lipoproteins (quadratic, P < 0.09). In these experiments, Cr propionate had no effect on overall growth performance, variable effects on carcass traits and plasma metabolites, and some positive effects on pork quality, especially water holding capacity of a fresh or frozen chop. PMID- 14552378 TI - In vitro fermentation characteristics of selected oligosaccharides by swine fecal microflora. AB - The objective of this study was to quantify the fermentation characteristics of oligosaccharides present in feed ingredients or isolated for dietary supplementation. Substrates studied included short-chain fructooligosaccharides, medium-chain fructooligosaccharides, long-chain fructooligosaccharides, raffinose, stachyose, soy solubles, granular and liquid forms of transgalactooligosaccharides, glucooligosaccharides, mannanoligosaccharides, and xylooligosaccharides. Three healthy pigs that had never received antibiotics served as sources of fecal inoculum. Each substrate was fermented in vitro; samples were taken at 0, 2, 4, 8, and 12 h, and pH change and short-chain fatty acid (SCFA) and gas production determined. Gas production at 12 h did not differ (P > 0.05) among all fructooligosaccharides, transgalactooligosaccharides, soy solubles, and xylooligosaccharides. Raffinose, stachyose, and raffinose + stachyose fermentation resulted in the greatest (P < 0.05) gas production at 12 h of all substrates tested. The rate of gas production was greatest (P < 0.05) for stachyose and least (P < 0.05) for glucooligosaccharides and mannanoligosaccharides. Substrate did not affect (P > 0.05) time to attain maximal rate of gas production. The pH at 12 h for all fructooligosaccharides and xylooligosaccharides did not differ (P > 0.05). The pH values at 12 h for raffinose, stachyose, and raffinose + stachyose were highest (P < 0.05) compared with all other substrates. Total SCFA production at 12 h was similar for all fructooligosaccharides and transgalactooligosaccharides, glucooligosaccharides, and soy solubles. Total SCFA production was greatest (P < 0.05) for xylooligosaccharides, stachyose, and raffinose + stachyose, and least (P < 0.05) for mannanoligosaccharides and raffinose. Stachyose fermentation resulted in the greatest (P < 0.05) rate and earliest time to attain maximal rate of SCFA production. All oligosaccharides studied were readily fermentable but varied in amount and type of SCFA produced. Fermentation of the pure forms of oligosaccharides contained in soy solubles resulted in greater gas production and higher pH compared with soy solubles. The oligosaccharides in the soy solubles matrix seemed to behave differently than their pure counterparts. The high rates of fermentation of most oligosaccharides tested indicate that they may serve as fermentable carbohydrate sources in the terminal small intestine or large intestine of swine. PMID- 14552380 TI - Comparison of the enzyme-hydrolyzed casein, guanidination, and isotope dilution methods for determining ileal endogenous protein flow in the growing rat and pig. AB - The objectives of the two studies were to determine whether the guanidination and isotope dilution methods applied both by labeling the animal (15N-infusion method) and by diet (15N-dilution method) give similar estimates of ileal endogenous lysine (EL) and endogenous nitrogen (EN) flows, respectively, to that of the enzyme-hydrolyzed casein (EHC) method in the growing pig and to determine whether the guandination and 15N-dilution methods give similar estimates of EL and EN flows, respectively, to that of the EHC method in the rat. For the first study, the test diet contained guanidinated and enzymatically hydrolyzed casein (molecular weight < 5,000 Da), which was also labeled with 15N. Rats (n = 30; mean BW 178 g) and pigs (n = 6; mean BW 19.2 kg) received a preliminary EHC-based diet for 7 d. The test diet was then given to the rats and pigs on d 8. Digesta were sampled from the terminal 20 cm of ileum of killed animals. The EL flows determined using the guanidination method were lower than those determined using the EHC method (means of 298 vs. 382, and 214 vs. 287 microg/g of DMI, in the rat and pig, respectively; P < 0.05 for the rat and P < 0.01 for the pig). The EN flows determined with the 15N-dilution method were lower than those determined using the EHC method (means of 1,034 vs. 1,942 and 1,011 vs. 1,543 microg/g of DMI, in the rat and pig, respectively, P < 0.001 for the rat and P < 0.05 for the pig). In the second study, pigs (n = 6; mean BW 27 kg) were continuously infused via the jugular vein with 15N-leucine for 11 d. The pigs received an EHC-based diet (molecular weight < 5,000 Da) during this 11-d period, after which digesta were sampled at the terminal ileum under anesthesia. The EN flow determined using the 15N-infusion method (mean of 1,971 microg/g DMI) was higher (P < 0.01) than that determined using the EHC method (mean of 1,233 microg/g DMI). The guanidination method gave a lower estimate of EL flow than did the EHC method in both the pig and rat. The 15N-dilution method also gave a lower estimate of EN flow than the EHC method in the pig and rat, and the 15N-infusion method gave a higher estimate of EN flow than the EHC method in the pig. PMID- 14552381 TI - Dietary galactooligosaccharides affect ileal and total-tract nutrient digestibility, ileal and fecal bacterial concentrations, and ileal fermentative characteristics of growing pigs. AB - The objective of this study was to evaluate dietary galactooligosaccharide (Gal OS) addition on swine nutrient digestibility, ileal and fecal bacterial populations, and ileal short-chain fatty acid (SCFA) production, and to determine their impact on ileal fermentative characteristics in vitro. Twelve T-cannulated pigs (BW = 25 kg) were fed a diet free of Gal OS for 21 d. On d 22, ileal digesta samples were collected for an in vitro fermentation experiment (Exp. 1). Substrates included: raffinose/stachyose combination (R + S), soy solubles (SS), and transgalactooligosaccharides (TOS). Also included were the non-OS components of SS and TOS. Nine pigs (three donors per treatment) served as ileal effluent donors. Each substrate was fermented in vitro for 6 h, and pH and SCFA and gas production were determined. Pigs then were allotted to three treatments: a Gal OS free control diet and the control diet with either 3.5% added Gal OS from SS or TOS. Diets, feces, and digesta samples collected weekly for 6 wk on d 6 (feces) and 7 (digesta) were analyzed for DM, OM, CP, and chromic oxide concentrations. Feces and ileal digesta were analyzed for bifidobacteria and lactobacilli populations. Ileal digesta samples were analyzed for SCFA. On d 64, a second in vitro fermentation experiment (Exp. 2) was conducted using ileal effluent from three pigs per treatment and the same substrates used in Exp. 1. In vivo results showed that ileal and total tract DM and OM digestion were decreased (P < 0.05) by addition of both SS and TOS to the diet. Ileal and total-tract N digestibilities were decreased (P < 0.05) by dietary addition of SS. Fecal bifidobacteria and lactobacilli were increased (P < 0.05) by addition of SS and TOS to the diet. Ileal propionate and butyrate concentrations were greater (P < 0.05) for pigs fed diets containing both sources of Gal OS. In vitro results showed that fermentation data were not affected by donor animal adaptation to treatment. For both in vitro experiments, gas and SCFA production were higher (P < 0.05) for R + S than for SS or TOS. Fermentation of R + S resulted in a higher pH (P < 0.05) than did SS or TOS. Fermentation of non-OS components of SS and TOS resulted in more (P < 0.05) gas and SCFA production, and pH values that did not differ (P > 0.05) compared to SS and TOS. The Gal OS used in this study were prebiotics, increasing beneficial bacteria in vivo and SCFA concentrations both in vivo and in vitro. PMID- 14552382 TI - Detection of corn intrinsic and recombinant DNA fragments and Cry1Ab protein in the gastrointestinal contents of pigs fed genetically modified corn Bt11. AB - Genetically modified corn has been approved as an animal feed in several countries, but information about the fate of genetically modified DNA and protein in vivo is insufficient. Genetically modified corn Bt11 is developed by inserting a recombinant DNA sequence encoding insecticidal Cry1Ab protein from Bacillus thuringiensis subsp. kurstaki. We examined the presence of corn intrinsic and recombinant cry1Ab gene by PCR, and the Cry1Ab protein by immunological tests in the gastrointestinal contents of five genetically modified corn Bt11-fed and five nongenetically modified corn-fed pigs. Fragments of corn zein (242 bp), invertase (226 bp) and of ribulose-1,5-bisphosphate carboxylase/ oxygenase genes (1,028 bp) were detected in the gastrointestinal contents of both Bt11 and nongenetically modified corn-fed pigs. Fragments of recombinant cry1Ab gene (110 bp and 437 bp) were detected in the gastrointestinal contents of the Bt11-fed pigs but not in the control pigs. Neither corn intrinsic nor cry1Ab gene fragments were detected in the peripheral blood by PCR. The gastrointestinal contents were positive for Cry1Ab protein by ELISA, immunochromatography, and immunoblot; however, these methods did not work for blood and precluded conclusions about any potential absorption of the protein. These results suggest that ingested corn DNA and Cry1Ab protein were not totally degraded in the gastrointestinal tract, as shown by their presence in a form detectable by PCR or immunological tests. PMID- 14552383 TI - Effect of boron supplementation of pig diets on the production of tumor necrosis factor-alpha and interferon-gamma. AB - Two experiments were conducted to determine the effects of dietary B on the production of cytokines following an endotoxin challenge. In both experiments, pigs were obtained from litters generated from sows fed low-B (control) or B supplemented (5 mg/ kg, as-fed basis) diets. In Exp. 1 and 2, 28 and 35 pigs, respectively (21 d old), remained with their littermates throughout a 49-d nursery phase and were fed either a control or B-supplemented diet. In Exp. 1, 12 pigs per treatment were moved to individual pens at the completion of the nursery phase and fed their respective experimental diet. On d 99 of the study, pigs were injected with 150 microg of phytohemagglutinin (PHA) to evaluate a local inflammatory response. Pigs receiving the B-supplemented diet had a decreased (P < 0.01) inflammatory response following PHA injection. Peripheral blood monocytes were isolated from six pigs per treatment on d 103 and cultured in the presence of lipopolysaccharide (LPS) to determine the effect of dietary B on tumor necrosis factor-alpha (TNF-alpha) production from monocytes. Isolated monocytes from pigs that received the B-supplemented diet had a numerically greater (P = 0.23) production of TNF-alpha. In Exp. 2, pigs were group housed with their littermates following the nursery phase for 43 d, after which 10 pigs per treatment were moved to individual pens. In Exp. 1 and 2, pigs were assigned randomly within dietary treatment to receive either an i.m. injection of saline or LPS on d 117 and d 109, respectively. The dose of LPS in Exp. 1 and 2 was 100 and 25 microg of LPS/kg of BW, respectively. In Exp. 1, serum TNF-alpha was increased (P < 0.01) at 2 h and tended to be increased (P < 0.11) at 6 and 24 h after injection by dietary B; however, only numerical trends existed for a B induced increase in TNF-alpha in Exp. 2. Serum interferon-gamma (IFN-gamma) was increased (P < 0.01) at 6 h and tended to be increased (P < 0.08) at 24 h after injection in Exp. 1. In Exp. 2, dietary B also numerically increased IFN-alpha. These data indicate that dietary B supplementation increased the production of cytokines following a stress, which indicates a role of B in the immune system; however, these data do not explain the reduction in localized inflammation following an antigen challenge in pigs. PMID- 14552384 TI - Sleep time following anesthesia in mouse lines selected for resistance or susceptibility to fescue toxicosis. AB - In previous work, a mouse line selected for resistance (R) to fescue toxicosis had higher activities of two hepatic Phase II detoxification enzymes than a mouse line selected for fescue toxicosis susceptibility (S). The primary objective of the present study was to determine whether those same lines also differed in hepatic Phase I enzyme activity, estimated from sleep time (ST) following sodium pentobarbital anesthesia. Additional objectives were to determine whether ST differences between lines were modulated by endophyte-infected fescue in the diet (with or without an enzyme inducer) and whether ST of individual mice was correlated with the effect of a toxin-containing diet on the postweaning growth of those mice. In Exp. I, 24 males from each line were randomly assigned to each of five diets: control (commercial rodent food meal); E+ (50% endophyte-infected fescue seed, 50% control); E+P (the E+ diet supplemented with 1,000 ppm phenobarbital); E- (50% endophyte-free fescue seed, 50% control); and E-P (the E- diet supplemented with 1,000 ppm phenobarbital). After 4 wk on these diets, ST was measured on all the mice. A second ST was recorded on each mouse by randomly sampling one-fourth of the population after 1, 2, 3, or 4 wk on a pelleted rodent food diet. Regardless of diet, R mice had shorter first and second ST than S mice (P < 0.01), suggesting higher hepatic Phase I microsomal enzyme activity. Mice on both phenobarbital-supplemented diets had shorter first ST than mice whose diets did not include that microsomal enzyme inducer (P < 0.01). In Exp. II, ST was measured on male and female R and S mice (n = 280) after they had been fed the E- diet for 2 wk, then the E+ diet for 2 wk, and then a pelleted rodent food diet for 2 wk. Growth response to the E+ diet was the percentage of reduction in gain on the E+ diet compared to gain on the E- diet the previous 2 wk. As in Exp. I, S mice slept longer than R mice (P < 0.01). The residual correlation between ST and gain reduction associated with the E+ diet equaled 0.04. Thus, an animal's apparent Phase I enzyme activity did not predict its growth rate depression on the toxin-containing diet. Based on these and previous studies, divergent selection for toxicosis response in mice was successful partially by causing divergence in activities of hepatic Phase I and II detoxification enzymes. PMID- 14552386 TI - Evaluation of the uterine environment and embryos of prepubertal gilts. AB - A series of three experiments was conducted to test the functional status of the uterus and embryos in prepubertal gilts. In Exp. 1, gilts were induced to ovulate by treating with gonadotropins followed by hCG 72 or 96 h later, and were artificially inseminated 24 h after hCG. Five of the 10 gilts treated at 120 d of age, but none of the gilts treated at 100 of age, maintained pregnancies. We next tested the function of the uterine environment by transferring embryos from postpubertal females into gilts of various ages that had been induced to ovulate but not inseminated (Exp. 2). Pregnancy rate at d 50 of gestation was 44% (4/9) for 100-d-old recipients, 67% (2/3) for 140-d-old recipients, and 60% (3/5) for postpubertal recipients (P > 0.20). Therefore, uteri of 100-d-old gilts are able to maintain pregnancies with conceptuses from postpubertal gilts. In Exp. 3, embryos from 100-d-old and postpubertal gilts were transferred into postpubertal recipients. Uterine horns of recipients were surgically separated before transfer, and embryos from 100-d-old and post-pubertal females were transferred to opposite horns of some recipients (experimental). Other recipients received embryos from postpubertal females in both uterine horns (control). When examined on d 50 to 60 of gestation, three of five control gilts were pregnant and three of seven experimental gilts were pregnant (P > 0.50). In experimental recipients, the survival of embryos from 100-d-old gilts was 38% (8/21) compared to 57% (15/26) for embryos from postpubertal gilts (P > 0.30). Because all uterine horns of pregnant recipients contained fetuses, these results support the hypothesis that embryos from 100-d-old gilts are able to initiate and maintain pregnancies in the uteri of postpubertal gilts. Therefore, the uterine environment of 100-d old gilts provides an environment that supports development of embryos produced by postpubertal gilts, and the embryos produced by 100-d-old gilts can survive and develop in the uteri of postpubertal gilts. It was only the combination of embryos and uteri of 100-d-old gilts that did not permit pregnancy to be maintained. PMID- 14552385 TI - Domperidone can ameliorate deleterious reproductive effects and reduced weight gain associated with fescue toxicosis in heifers. AB - The aim of this study was to evaluate the effect of a dopamine antagonist, domperidone, in nonpregnant, reproductively cycling heifers consuming endophyte infected (EI) fescue diets. Thirty crossbred heifers (Angus x Holstein or Hereford x Holstein) were assigned to one of three treatment groups (n = 10); endophyte-free (EF) fescue diet, EI fescue diet, or endophyte-infected diet and treated with domperidone (EID). Heifers fed EI diets had decreased weight gains compared with heifers fed EF or EID (P < 0.05) during a 21-d treatment period. Ovarian structures were monitored via transrectal ultrasound to determine follicle size and day of ovulation. Blood plasma samples were collected daily and analyzed for progesterone concentration to determine luteal function. Heifers ingesting EI diets had estrous cycles of shorter duration and lower mid-cycle progesterone concentrations than heifers in the EF or EID treatments (P < 0.05). Ovaries from a subset of heifers in each group (n = 3 per group) were harvested and in vitro secretion of progesterone from luteal tissue extracts was determined. No differences in progesterone concentrations were detected among luteal tissue incubates (P > 0.05). These results suggest that domperidone supplementation of heifers consuming EI fescue may ameliorate certain symptoms of fescue toxicosis. PMID- 14552387 TI - Quantification of mammary gland tissue size and composition changes after weaning in sows. AB - The objectives of this study were to characterize the tissue compositional changes in porcine mammary glands after weaning and to determine whether administration of estradiol alters the profile of these tissue changes. Forty five primiparous sows were assigned randomly to one of two treatment groups after weaning, control or estrogen treated. Estrogen-treated sows received twice-daily injections of estradiol-17beta (0.125 mg/kg of BW); control sows received vehicle injections. Sows were weaned at d 21 of lactation and killed on either d 0 (d of weaning; n = 5) or on d 2, 3, 4, 5, or 7 after weaning (n = 4 per treatment on each day). Teat order relative to suckling behavior was observed on the day before weaning to determine which mammary glands the piglets suckled. Suckled and non-suckled glands were identified from the teat order observation, and individual mammary glands were collected at slaughter. Mammary glands were trimmed of skin and extraneous fat pad, individually weighed, and bisected to measure cross-sectional area. The remaining half of each gland was ground and stored at -20 degrees C for chemical analyses. Frozen tissue was used for measuring tissue DNA, DM, protein, fat, and ash contents. Suckled mammary glands of sows undergo significant and dramatic changes during the initial 7 d after weaning, with significant changes occurring even by d 2 after weaning. Mean cross sectional area of parenchymal tissue in suckled mammary glands decreased from 59.7 +/- 2.1 cm2 on the day of weaning to 26.8 +/- 2.3 cm2 by d 7 after weaning (P < 0.0001). Mammary gland wet weight decreased from 485.9 +/- 22.0 g on the day of weaning to 151.5 +/- 24.8 g by d 7 after weaning (P < 0.0001), whereas DNA decreased from 838.8 +/- 46.2 g on the day of weaning to 278.4 +/- 52.5 g by d 7 after weaning (P < 0.0001). The changes in gland wet weight and DNA during the period of mammary gland involution in the sow represent loses of over two-thirds of the parenchymal mass and nearly two-thirds of the cells that were present on the day of weaning. Estrogen treatment did not affect overall mammary involution during the first 7 d after weaning. Mammary glands that were not suckled during lactation had no further loss of parenchymal tissue during the first 7 d after weaning. Mammary gland involution in the sow is a rapid process and is probably irreversible within 2 or 3 d after weaning. PMID- 14552388 TI - Leptin, tumor necrosis factor-alpha (TNF), and CD14 in ovine adipose tissue and changes in circulating TNF in lean and fat sheep. AB - Four studies were designed to determine whether 1) tumor necrosis factor-alpha (TNF) and the Lipopolysaccharide (LPS) binding ligand, CD14, are produced by sheep adipose tissue; 2) nutritional reserves and/or short-term fasting affect circulating concentrations of TNF; 3) there is a relationship between TNF and metabolic factors in sheep; and 4) inflammation alters circulating concentrations of leptin. In Exp. 1 and 2, ewes were assigned, based on ultrasonic assessments of last-rib subcutaneous fat measurements to fat (fat thickness > 1 cm; mean = 1.52 +/- 0.03 cm) or thin (fat thickness < 1 cm; mean = 0.25 +/- 0.03 cm) groups. Fat and thin ewes were assigned to fed or fasted groups for a total of four groups (fed-fat; fasted-fat; fed-thin; fasted-thin). Fed-ewes had ad libitum access to feed, and fasted-ewes were prohibited feed 48 h before initiation of sample collection. In Exp. 1, subcutaneous fat samples were collected from just above the last rib for detection of TNF and CD14 mRNA, and immunoreactivity. Tumor necrosis factor-alpha-like immunoreactivity in adipocytes was sparse, more pronounced in cells in fed-ewes than fasted-ewes, and localized to membranes between adjacent cells in nucleated regions. Immunoreactivity for CD14 was minimally observed but present in adipocytes and widely expressed in infiltrating monocytes and epithelial vascular cells. Leptin was detected in adipocytes. In Exp. 2, plasma samples collected every 6 h for 24 h were analyzed for plasma concentrations of TNF. Fat ewes had greater plasma concentrations of TNF than thin ewes (P = 0.039). In Exp. 3, wethers were injected i.v. with interleukin 1beta or TNF. Blood samples were collected every 15 min for 8 h following injection. Plasma concentration of leptin was not affected by treatment (P > 0.39). In Exp. 4, wethers were injected with LPS. Blood samples were collected every 15 min for 8 h following injection. Plasma concentration of leptin was not altered by LPS (P > 0.20). These results provide evidence: 1) of TNF-like immunoreactivity within fat tissue; 2) that elements within fatty tissues have CD14 that may allow adipocyte function to be directly affected by LPS; 3) that plasma concentrations of leptin are not altered by LPS treatment; and 4) that circulating concentrations of TNF are elevated with obesity in sheep. PMID- 14552389 TI - Effect of feeding glyphosate-tolerant (roundup-ready events GA21 or nk603) corn compared with reference hybrids on feedlot steer performance and carcass characteristics. AB - Three experiments were conducted to compare the feeding value of genetically enhanced corn (Roundup Ready corn events GA21 and nk603) with nontransgenic hybrids. The four treatments included two separate reference hybrids (REF), the near-isogenic control hybrid (CON), and the genetically enhanced corn (RR), resulting in two preplanned comparisons of CON vs. RR and RR vs. the average of REF. In Exp. 1 (RR event GA21), 175 steers (BW = 427 kg) were fed in 25 pens with seven pens per corn hybrid, except CON, which contained four pens due to limited quantities of that hybrid. In Exp. 2 (RR event nk603), 196 steers (BW = 420 kg) were fed in 28 pens with seven pens per corn. In Exp. 3 (RR event nk603), 200 steers were fed in 20 pens, with a similar treatment design to Exp. 2 and five pens per corn. All experiments were conducted as completely randomized designs and utilized corn produced at University of Illinois (Exp. 1 and 2) and University of Nebraska (Exp. 3) research farms under identity-preserved protocols. In all experiments, DMI, ADG, and feed efficiency were similar (P > 0.30) between RR and REF. In Exp. 1 and Exp. 2, RR was not different (P > 0.25) than CON for growth performance. In Exp. 3, RR was not different from CON for ADG and DMI (P > 0.15) or for feed efficiency (P = 0.08). No differences were observed between RR and CON or RR and REF for carcass weight, longissimus dorsi area, and marbling scores in any of the experiments. Subtle differences were observed between RR and either CON or REF for fat depth in each experiment; however, cattle fed RR were not consistently greater and varied from either the CON or the REF (but not both contrasts) within an experiment. Based on these results, insertion of glyphosate-tolerant genes had no significant effect on nutritive quality of corn. Performance and carcass characteristics were not influenced, which suggests that Roundup Ready corn is similar to conventional, nontransgenic corn when fed to finishing feedlot cattle. PMID- 14552390 TI - Effects of volatile fatty acid supply on their absorption and on water kinetics in the rumen of sheep sustained by intragastric infusions. AB - Three sheep fitted with a ruminal cannula and an abomasal catheter were used to study water kinetics and absorption of VFA infused continuously into the rumen. The effects of changing VFA concentrations in the rumen by shifting VFA infusion rates were investigated in an experiment with a 3 x 3 Latin square design. On experimental days, the animals received the basal infusion rate of VFA (271 mmol/h) during the first 2 h. Each animal then received VFA at a different rate (135, 394, or 511 mmol/h) for the next 7.5 h. Using soluble markers (polyethylene glycol and Cr-EDTA), ruminal volume, liquid outflow, apparent water absorption, and VFA absorption rates were estimated. There were no significant effects of VFA infusion rate on ruminal volume and water kinetics. As the VFA infusion rate was increased, VFA concentration and osmolality in the rumen were increased and pH was decreased. There was a biphasic response of liquid outflow to changes in the total VFA concentration in the rumen, as both variables increased together up to a total VFA concentration of 80.1 mM, whereas, beyond that concentration, liquid outflow remained stable at an average rate of 407 mL/h. There were significant linear (P = 0.003) and quadratic (P = 0.001) effects of VFA infusion rate on the VFA absorption rate, confirming that VFA absorption in the rumen is mainly a concentration-dependent process. The proportion of total VFA supplied that was absorbed in the rumen was 0.845 (0.822, 0.877, and 0.910 for acetate, propionate, and butyrate, respectively). The molar proportions of acetate, propionate, and butyrate absorbed were affected by the level of VFA infusion in the rumen, indicating that this level affected to a different extent the absorption of the different acids. PMID- 14552392 TI - Screening of exogenous enzymes for ruminant diets: relationship between biochemical characteristics and in vitro ruminal degradation. AB - With the objective of developing a rational approach for the selection of feed enzymes for ruminants, 22 commercial enzyme products were examined in terms of protein concentration, enzymic activities on model substrates, and hydrolytic capacity, the latter determined from the release of reducing sugars from alfalfa hay and corn silage. An in vitro ruminal degradation assessment was carried out using the same substrates, untreated or treated with the 22 enzyme products at 1.5 microL/g forage DM. Stepwise regressions were then performed to establish relationships between these factors. Protein concentration and enzymic activities explained at least 84% (P < 0.01) of the variation in the release of reducing sugars from alfalfa and corn silage. Alfalfa DM degradation after incubation with ruminal fluid for 18 h was positively related to xylanase activity (R2 = 0.29, P < 0.01), but the same activity was negatively related to DM degradation of corn silage (R2 = 0.19, P < 0.05). Protease activity explained a further 10% of the alfalfa DM degradation (P < 0.10). Following sequential steps involving the determination of rate and extent of DM and fiber degradation, the best candidates for alfalfa and corn silage were selected. Enzyme products effective with alfalfa hay seemed to exert part of their effect during the pretreatment period, whereas enzymes effective with corn silage worked exclusively after ruminal fluid was added. This finding suggests that different modes of action of exogenous enzymes are attacking different substrates and may partly explain enzyme-feed specificity. In alfalfa, it seems that effective enzymes work by removing structural barriers that retard the microbial colonization of digestible fractions, increasing the rate of degradation. In corn silage, effective enzymes seem to interact with ruminal enzymes to degrade the forage more rapidly, which is consistent with previous findings of synergism between exogenous and ruminal enzymes. PMID- 14552391 TI - Effects of enzyme supplementation of a total mixed ration on microbial fermentation in continuous culture, maintained at high and low pH. AB - A dual-flow continuous culture system was used to investigate the effects of pH and addition of an enzyme mixture to a total mixed ration (TMR) on fermentation, nutrient digestion, and microbial protein synthesis. A 4 x 4 Latin square design with a factorial arrangement of treatments was used, with four 9-d periods consisting of 6 d for adaptation and 3 d for measurements. Treatments were as follows: 1) high pH with control TMR, 2) high pH with TMR treated with enzyme, 3) low pH with control TMR, and 4) low pH with TMR treated with enzyme. Ranges of pH were 6.0 to 6.6 and 5.4 to 6.0 for high and low, respectively. Fermenters were fed twice daily a TMR consisting of 30% alfalfa hay, 30% corn silage, and 40% rolled corn (DM basis). The silage was milled fresh and the TMR was fed to the fermenters in fresh form (64% DM). The enzyme mixture was a commercial product of almost exclusive protease activity; it was applied daily to the fresh TMR and stored at 4 degrees C for at least 12 h before feeding. Degradability of OM, NDF, ADF, and cellulose was decreased (P < 0.05) by low pH. Hemicellulose and protein degradation were not affected by pH. Enzyme addition increased (P < 0.01) NDF degradability (by 43% and 25% at high and low pH, respectively), largely as a result of an increase in hemicellulose degradation (by 79% and 51% at high and low pH, respectively). This improvement was supported by an increase (P < 0.05) in the xylanase and cellulase activities in the liquid phase of the fermenter contents. Total VFA were decreased (P < 0.05) by low pH, but were not affected by enzyme addition. Total bacterial numbers were increased (P < 0.03) at low pH and tended (P < 0.13) to increase with enzyme addition. Cellulolytic bacteria in effluent fluid were decreased (P < 0.02) at low pH but were unaffected by enzyme addition. Despite a large increase (P < 0.001) in protease activity, protein degradation was only numerically increased by enzyme addition. Microbial protein synthesis was higher (P < 0.10) at high pH but was not affected by enzyme addition. Methane production, expressed as a proportion of total gases, was decreased (P < 0.001) at low pH but was not affected by enzyme addition. It is concluded that it is possible to adapt the continuous culture system to use fresh feeds instead of dried feeds. Overall, the results indicate that the enzyme product used in this study has a potential to increase fiber degradability without increasing methane production. PMID- 14552393 TI - Composition and digestive tract retention time of ruminal particles with functional specific gravity greater or less than 1.02. AB - The objective of this study was to determine composition, particle size distribution, and in vivo kinetics of ruminal particles having functional specific gravity (FSG) greater or less than FSG of particles found in the omasum and reticulum of lactating dairy cows. Particles from the reticulum and the omasal had FSG of 1.03 and 1.02, respectively. Particles from ruminal contents with FSG higher (HP) or lower (LP) than 1.02 were isolated and labeled with Er or Dy, respectively. Four ruminally cannulated, lactating Ayrshire dairy cows were fed all-grass silage (AS) or 54% grass silage:46% concentrate (SC) diets in a cross-over design trial and used to study chemical composition and ruminal and total tract kinetics of HP and LP. Labeled particles were pulse dosed into the rumen of the cows and disappearance of the markers from ruminal HP and LP pools and excretion in feces was monitored for 72 and 120 h, respectively. Fecal marker excretion data were fitted using two-compartment mathematical age-dependent/age independent (Gn-->G1) models. Inclusion of concentrate in the diet (SC) increased (P < 0.05) apparent total tract digestibility of dietary DM, OM and N. Digestibility of fiber fractions, NDF and ADF, was lower (P < 0.01 and P < 0.05, respectively) for SC compared with AS. The heavy particles had higher (P < 0.01) indigestible NDF and lower (P < 0.01) N concentration than LP. Particles from the HP pool passed from the rumen more rapidly (P < 0.01) than particles from LP (0.044 and 0.019 h(-1), respectively). Diet had no effect on particle rate of disappearance or pool size in the rumen. Across diets, pool size of LP was consistently larger (P < 0.05) than that of HP. Diet had no effect on total tract mean retention time (MRT) of LP or HP. Total tract MRT of LP was greater (P < 0.05) than MRT of HP (59.6 vs. 49.0 h, respectively). Results from this study support the hypothesis that functional specific gravity is an important factor determining the rate of outflow and residence time of feed particles within the reticulo-rumen and total digestive tract. Our data indicate that digesta particles with functional specific gravity greater or less than 1.02 have different composition and flow characteristics. Heavier particles contain more indigestible fiber and less N and are likely depleted of substrate available for microbial fermentation, are smaller in size, and have a higher passage rate/shorter retention time in the digestive tract than lighter particles. PMID- 14552394 TI - Nitrogen losses in metabolism trials. PMID- 14552395 TI - Studying the effects of early child care experiences on the development of children of color in the United States: toward a more inclusive research agenda. AB - Evidence is presented of the different cultural and ecological contexts affecting early child care for families of color. It is argued that improvements on previous research require a fundamental shift in how race, ethnicity, and culture as psychological variables are examined. Furthermore, to avoid the pitfalls and failures of previous research, new research must incorporate expanded models of child care and development in childhood. The integrative model of development for children of color proposed by Garcia Coll et al. (1996) is presented as a basis for developing more specific ecological models relevant to addressing child care issues in ethnic minority families. Finally, priority areas for future research are recommended to stimulate and enable child care researchers to adopt a more inclusive view of child care and its effects. PMID- 14552396 TI - The development of argument skills. AB - This work sought to obtain experimental evidence to corroborate cross-sectional patterns of development in argument skills and to evaluate the effectiveness of an intervention designed to foster development of these skills in academically at risk 13- to 14-year-olds. Students participated in 16 sessions of a collaborative, goal-based activity providing dense exercise of argumentive thinking. One condition included peer dialogues; another did not. The former was the more effective, although both groups progressed. Participants showed increased frequency of usage of powerful argumentive discourse strategies, such as counterargument, and decreased frequency of less effective strategies. Quality of individual arguments (for or against a claim) also improved, supporting the existence of a close relation between these two kinds of argument skills. PMID- 14552397 TI - Sibling differentiation: sibling and parent relationship trajectories in adolescence. AB - Studied here were the links between sibling differences in trajectories of change in the qualities of parent-child relationships and the qualities of sibling relationships across a 2-year period in adolescence. Participants were first- and second-born siblings (M age = 14.94 years for firstborns and M age = 12.46 years for secondborns) from 185 predominantly White, working and middle-class families. In home interviews, siblings reported on their dyadic family relationships. For reports of parent-child warmth but not parent-child conflict, results were consistent with sibling differentiation theory: Increasing differences between siblings over time in parent-child warmth were linked to trajectories of increasing warmth and decreasing conflict in the sibling relationship as reported by firstborns, and increasing warmth in the sibling relationship as reported by secondborns. The findings support the view that sibling differentiation may be a strategy for managing sibling conflict and rivalry. PMID- 14552398 TI - Overcoming the positive-capture strategy in young children: learning about indeterminacy. AB - Two experiments were conducted to examine whether and how 4- and 5-year-olds learn to distinguish determinate from indeterminate evidence. Children were asked to decide whether various patterns of evidence were sufficient to reach unambiguous conclusions. This study replicated the finding that young children tend to use a strategy that, although generally successful, fails on evidence patterns in which a single positive instance co-occurs with an unexplored source of evidence. Experiment 1 demonstrated that this positive-capture strategy is deeply entrenched, even in a meaningful, pragmatic context. With a microgenetic design, Experiment 2 revealed that young children are capable of replacing the positive-capture strategy with a correct strategy when they are exposed to various analogous tasks in several training sessions. PMID- 14552399 TI - Do young children always say yes to yes-no questions? A metadevelopmental study of the affirmation bias. AB - The present study investigated whether yes-no questions would lead to a yes bias in young children. Four experiments were conducted in which 2- to 5-year-olds were asked comprehensible and incomprehensible yes-no questions concerning familiar and unfamiliar objects. Consistent findings were obtained: (a) 2-year olds displayed a consistent yes bias; (b) 4- and 5-year-olds exhibited no response bias toward comprehensible questions and a nay-saying bias toward incomprehensible questions; and (c) 3-year-olds' results were mixed, suggesting that the age of 3 years is a period of developmental transition in response tendency toward yes-no questions. The findings suggest that yes-no questions are suitable for older children, providing they are comprehensible, but may result in biased results when used with younger children and when incomprehensible. PMID- 14552400 TI - Prosocial development in relation to children's and mothers' psychological problems. AB - The study tested whether children's prosocial behavior was negatively or positively related to children's and mothers' psychological problems. Participants were 149 London families when mothers were pregnant and followed up in infancy and at ages 4 and 11. Children's cooperation at 4 and general prosocial tendencies at 11 were negatively associated with externalizing problems but unrelated to internalizing problems. A subgroup of children who were more prosocial than average expressed clinically significant worries about family members. Maternal depression decreased prosocial behavior in the eyes of adults, but children of depressed mothers saw themselves to be prosocial. Early cooperation protected children against later risk for externalizing problems, even when their early behavioral problems were taken into account. PMID- 14552401 TI - Peer relations across contexts: individual-network homophily and network inclusion in and after school. AB - Peer relations across 2 contexts (in school and after school) were examined for 577 participants, approximately 12 years old, from 3 middle schools in Milan, Italy. The primary research questions were: Do peer networks from different contexts uniquely contribute to explaining variance in individual behavior? Do measures of peer preference and peer network inclusion across contexts uniquely contribute to explaining individual depressive symptoms? Structural equation models showed that both the in-school and the after-school peer networks uniquely contributed to explaining variance in 2 types of individual problem behavior (in school problem behavior, after-school delinquency), and that similarity with the 2 peer networks varied according to behaviors specific to each context and across gender. Finally, both in-school and after-school peer network inclusion contributed to explaining variance in depressive symptoms, after controlling for classroom peer preference. PMID- 14552402 TI - The role of chronic peer difficulties in the development of children's psychological adjustment problems. AB - A longitudinal investigation was conducted to explicate how the confluence of early behavioral dispositions, relational histories, and cognitive representations of the self and others contributes to internalizing problems, externalizing problems, and loneliness. One-hundred and ninety three girls, and 206 boys were assessed annually from age 5 (kindergarten) to age 10 (Grade 4). Early aggressive behavior was related to Grade 4 maladjustment directly and indirectly through subsequent relational stressors. Significant associations emerged between chronic friendlessness and rejection and later adaptation not accounted for by concurrent relational difficulties. Self- and peer beliefs partially mediated the relation between peer difficulties and internalizing problems and loneliness. The results highlight the utility of child-by environment models as a guide for the investigation of processes that antecede psychosocial maladjustment. PMID- 14552403 TI - The specificity of environmental influence: socioeconomic status affects early vocabulary development via maternal speech. AB - The hypothesis was tested that children whose families differ in socioeconomic status (SES) differ in their rates of productive vocabulary development because they have different language-learning experiences. Naturalistic interaction between 33 high-SES and 30 mid-SES mothers and their 2-year-old children was recorded at 2 time points 10 weeks apart. Transcripts of these interactions provided the basis for estimating the growth in children's productive vocabularies between the first and second visits and properties of maternal speech at the first visit. The high-SES children grew more than the mid-SES children in the size of their productive vocabularies. Properties of maternal speech that differed as a function of SES fully accounted for this difference. Implications of these findings for mechanisms of environmental influence on child development are discussed. PMID- 14552404 TI - Strengthening sociometric prediction: scientific advances in the assessment of children's peer relations. AB - This study assessed the strength of sociometric classification in the prediction of concurrent sociobehavioral adjustment. Differential adjustment for subgroups of unclassified children were also examined. Participants were 881 fifth graders (ages 9 to 12). Classification strength (CS) and unclassified subgroups were determined through newly developed algorithms. CS added significantly to the prediction of all areas of adjustment. For example, highly rejected children were at extreme risk for victimization whereas highly controversial children were most likely to be bullies and relationally aggressive. Unclassified subgroups were found to exhibit adjustment problems mirroring those of their extreme status group counterparts. Findings support that increasing the sensitivity of sociometric measurement results in both greater predictive strength and enhanced understanding of underlying social processes. PMID- 14552405 TI - Testing the dynamic field theory: working memory for locations becomes more spatially precise over development. AB - The dynamic field theory predicts that biases toward remembered locations depend on the separation between targets, and the spatial precision of interactions in working memory that become enhanced over development. This was tested by varying the separation between A and B locations in a sandbox. Children searched for an object 6 times at an A location, followed by 3 trials at a B location. Two- and 4 year-olds', but not 6-year-olds', responses were biased toward A when A and B were 9-in. and 6-in. apart. When A and B were separated by 2 in., however, 4- and 6-year-olds' responses were biased toward A. Thus, the separation at which responses were biased toward A decreased across age groups, supporting the predictions of the theory. PMID- 14552406 TI - Young children's reasoning about many-to-one correspondences. AB - Young children's understanding of many-to-one correspondence problems was studied to illuminate the developmental transition from additive to multiplicative numerical knowledge. A many-to-one correspondence exists when a fixed number of target objects (greater than 1) is associated with each of a set of referents, as in putting 3 flowers in each of several vases. Two experiments examined effects of a brief training procedure that highlighted the iterative nature of many-to one mappings. In Experiment 1, 5- and 6-year-old children did not benefit from the training, but a subset of 7-year-olds did. In Experiment 2, 7-year-olds showed training effects that extended to generalization problems. Patterns of performance across experimental and generalization problems suggested that some children had difficulty applying what they learned from training to the experimental problems. PMID- 14552407 TI - The role of mental health factors and program engagement in the effectiveness of a preventive parenting program for Head Start mothers. AB - Head Start centers were randomly assigned to intervention (parent training) or control conditions, and the role of maternal mental health risk factors on participation in and benefit from parent training was examined. Parenting was measured by parent report and independent observation in 3 domains: harsh/negative, supportive/positive, inconsistent/ineffective parenting. Structural equation modeling showed that parent engagement training was associated with improved parenting in a dose-response fashion. Mothers with mental health risk factors (i.e., depression, anger, history of abuse as a child, and substance abuse) exhibited poorer parenting than mothers without these risk factors. However, mothers with risk factors were engaged in and benefited from the parenting training program at levels that were comparable to mothers without these risk factors. PMID- 14552408 TI - Modeling the impacts of child care quality on children's preschool cognitive development. AB - The National Institute of Child Health and Human Development (NICHD) Study of Early Child Care compared 3 statistical methods that adjust for family selection bias to test whether child care type and quality relate to cognitive and academic skills. The methods included: multiple regression models of 54-month outcomes, change models of differences in 24- and 54-month outcomes, and residualized change models of 54-month outcomes adjusting for the 24-month outcome. The study was unable to establish empirically which model best adjusted for selection and omitted-variable bias. Nevertheless, results suggested that child care quality predicted cognitive outcomes at 54 months, with effect sizes of .04 to .08 for both infant and preschool ages. Center care during preschool years also predicted outcomes across all models. PMID- 14552409 TI - Theory of mind and relational complexity. AB - Cognitive complexity and control theory and relational complexity theory attribute developmental changes in theory of mind (TOM) to complexity. In 3 studies, 3-, 4-, and 5-year-olds performed TOM tasks (false belief, appearance reality), less complex connections (Level 1 perspective-taking) tasks, and transformations tasks (understanding the effects of location changes and colored filters) with content similar to TOM. There were also predictor tasks at binary relational and ternary-relational complexity levels, with different content. Consistent with complexity theories: (a) connections and transformations were easier and mastered earlier than TOM; (b) predictor tasks accounted for more than 80% of age-related variance in TOM; and (c) ternary-relational items accounted for TOM variance, before and after controlling for age and binary-relational items. Prediction did not require hierarchically structured predictor tasks. PMID- 14552410 TI - Effects of earnings-supplement policies on adult economic and middle-childhood outcomes differ for the "hardest to employ". AB - Data from the Minnesota Family Investment Program and the New Hope demonstration were used to determine whether experimental effects of antipoverty policies differ by parents' risk for nonemployment. Using propensity score analysis, increases in employment and income were largest in the harder-to-employ halves of both samples. However, only children in the moderately hard-to-employ quartiles (50th to 75th percentile) consistently showed improvements in school and behavior outcomes. The very-hardest-to-employ 25% experienced decreases in school engagement, and increases in aggressive behaviors, despite substantial increases in parental employment and income. In this group, increases in maternal depression, reductions in regular family routines, and smaller increases in job stability and center-based child care occurred. These factors may have counteracted the potential benefits of increased income on children. PMID- 14552411 TI - Perceptions and functions of play and real fighting in early adolescence. AB - The hypothesis is tested that adolescent boys' (mean age of 12.8 years) intrasexual rough-and-tumble play (R&T) is used for dominance and intersexual R&T is used to establish heterosexual relationships. In Study 1, boys' observed R&T was related to both dominance and aggression. In the first half of the school year, R&T occurred primarily between males, possibly to establish dominance. In the second half of the year, both boys and girls engaged in R&T, possibly to establish heterosexual relationships. Counter to the hypothesis, observed aggression increased across the year. In Study 2, youngsters viewed taped R&T bouts in which they were participants or nonparticipants. Participant, more than nonparticipant, males saw R&T as related to dominance whereas participant, more than nonparticipant, females saw it as playful. PMID- 14552412 TI - Infant stress and parent responsiveness: regulation of physiology and behavior during still-face and reunion. AB - This study examined infant response and recovery from a social challenge and parent responses. Behavioral and physiological responses were measured from forty three 5- and 6-month-olds infants during a modified still-face procedure that used an additional still-face reunion sequence. Results confirm the hypothesis that infants of more responsive parents show more regulation than infants of less responsive parents. Infants of more responsive parents showed greater regulation of heart rate and negative affect during the final episode of the procedure than infants of less responsive parents. In addition, this procedure elicited a cortisol response (from .22 microg/dl to .31 microg/dl). Findings suggest important links between parent behavior and infant stress reactivity and regulation. PMID- 14552413 TI - Preschoolers' use of form class cues to learn descriptive proper names. AB - This study examined 3- and 4-year-old preschoolers' ability to learn proper names containing familiar descriptions. Children saw a novel creature with a familiar property (it was red) and heard either an adjective ("This is a red one") or a descriptive proper name ("This is Mr. Red"). The creature was then transformed, losing the property (e.g., it became green). Children had to extend the word to either the transformed original creature or a new creature bearing the original property (another red creature). Children, especially 4-year-olds, extended the adjective to the new creature but were significantly more likely to extend the proper name to the original creature. Lexical form class cues provided potent information about word meaning, directing preschoolers to reinterpret familiar descriptive terms (adjectives) as homophonic terms designating unique individuals (proper names). PMID- 14552414 TI - Community violence exposure, social cognition, and aggression among urban elementary school children. AB - The effects of witnessing community violence on aggressive cognitions and behavior were investigated in an ethnically diverse sample of 4,458 children living in urban neighborhoods. Prior violence exposure had a significant effect in increasing aggression, normative beliefs about aggression, and aggressive fantasy. Although exposure to violence predicted aggressive behavior both in Grades 1 through 3 (ages 5-8) and Grades 4 through 6 (ages 9-12), the effects on social cognition were only evident in the later grades. Furthermore, the effect of violence exposure on aggression in the later grades was partially mediated by its effect on social cognition. These findings suggest that witnessing community violence has an effect on children's aggressive behavior through both imitation of violence and the development of associated cognitions as children get older. PMID- 14552415 TI - A peptide centenary celebration. PMID- 14552416 TI - Solid-phase synthesis: a paradigm shift. AB - A personal review by the first graduate student of Professor R. Bruce Merrifield of the evolution of solid-phase synthesis and its acceptance by various subsets of the chemistry community. Solid-phase synthesis, as currently practised in the synthesis of biopolymers, combinatorial solid-phase organic chemistry, synthesis of natural products, catalyst selection, chemical ligation and materials development, has proven a paradigm shift for the chemistry community. PMID- 14552417 TI - The fluorenylmethoxycarbonyl group in solid phase synthesis. AB - The history of the fluorenylmethoxycarbonyl amino-protecting group since its introduction into solid phase peptide synthesis in 1978 is briefly traced. PMID- 14552418 TI - What to synthesize? From Emil Fischer to peptidomics. AB - The driving forces, incentives and strategic targets of peptide synthesis have undergone considerable evolution during the centenary following the pioneer work of Emil Fischer. In those days peptide synthesis was considered as a way of confirming the polypeptide theory of protein structure. The scientific community also expected (naively) that the synthesis would eventually lead to the creation of artificial living organisms. Only in the 1950s, when the first exact amino acid sequences were established did peptide chemistry obtain firmer ground and clearly defined targets. The total synthesis of peptide hormones and antibiotics became possible, providing valuable material for elucidating structure-functional relationships and the mechanisms of biological action. In the following years the number of peptides isolated from various biological sources grew with impressive speed and peptides became known as the most abundant, ubiquitous group of low molecular bioregulators. The design and synthesis of novel peptide based pharmaceuticals became an important area of peptide chemistry. At present we are facing the challenge of analysing the structures and bioactivities of total sets of peptides, i.e. peptidoms, present in concrete tissues or groups of cells. The results obtained along these lines at the IBCH RAS Institute of Bioorganic Chemistry are briefly considered in the review. PMID- 14552419 TI - Synthetic glycopeptides for the development of tumour-selective vaccines. AB - Based on structural information reported for the tumour-associated epithelial mucin MUC1, glycopeptides have been synthesized which contain tumour-associated saccharide antigens. such as the Thomsen-Friedenreich (T), TN or sialyl TN antigen. in combination with peptide sequences of the tandem repeat region of MUC1. Solid-phase syntheses have been carried out using N-Fmoc protected O glycosyl serine and threonine building blocks and an allylic anchor which is stable to basic and acidic conditions, but can be cleaved under neutral conditions in a palladium(0)-catalysed allyl transfer reaction. In addition. a (2 3)sialyl T antigen threonine building block was prepared by a chemoenzymatic strategy and used in the synthesis of an N-terminal glycopeptide antigen of leukosialin (CD43). The proliferation of cytotoxic T cells could be induced using a construct consisting of a MUC1-glycopeptide antigen and a T cell epitope. PMID- 14552420 TI - Total chemical synthesis of enzymes. AB - The total synthesis, at will, of a wide variety of protein and enzyme molecules is made feasible by modem chemical ligation methods. As Emil Fischer intuitively understood, synthetic access to the enzyme molecule enables the power of chemical science to be applied to elucidating the molecular basis of catalytic function in unprecedented detail. PMID- 14552421 TI - The bold legacy of Emil Fischer. AB - A century has passed since Emil Fischer won the Nobel Prize in chemistry. From his first synthesis of glycyl-glycine in 1901 he has been a luminary to peptide chemists over the past 100 years. In this paper, a brief summary of some of the major accomplishments in peptide chemistry will be covered followed by a description of several of our own endeavours in peptide chemistry which arose from the discoveries of the giants of our field. We will include the development of a novel activating agent (DEPBT), the synthesis of a novel building block, alpha-methyl-D-cysteine, its incorporation into biologically active opioids, and conclude with the synthesis of dendritic collagen mimetics. PMID- 14552423 TI - Mouse tales from Kresge: the deafness mouse. AB - Mouse models for human deafness have not only proven instrumental in the identification of genes for hereditary hearing loss, but are excellent model systems in which to examine gene function as well as the resulting pathophysiology. One mouse model for human nonsyndromic deafness is the deafness (dn) mouse, a spontaneous mutation in the curly-tail (ct) stock. The dn gene is on mouse Chromosome 19 and it was recently shown to be a novel gene called Tmc1. A mutation in Tmc1 is also found in Beethoven (Bth), which is another deaf mouse mutant. In humans, one autosomal dominant form of nonsyndromic hearing loss (DFNA36) and two autosomal recessive forms (DFNB7 and DFNB11) are associated with mutations in TMC1, the human homologue of Tmc1. The transmembrane protein encoded by this gene is required for normal cochlear hair cell function and the mouse models will facilitate the elucidation of the molecular pathway that is disrupted when mutations are present. PMID- 14552422 TI - Localization of the P2Y4 receptor in the guinea pig organ of Corti. AB - Cochleae of guinea pigs were evaluated for the presence of the metabotropic receptor, P2Y4. Evidence is presented that P2Y4 protein is expressed in the guinea pig cochleae using Western blot analysis. A single protein band of 35 kDa was detected with P2Y4 receptor-specific antibody. The cellular distribution of P2Y4 purinoceptor protein was determined by immunohistochemistry of the whole organ of Corti. Immunoreactive staining for P2Y4 was seen in most cells of the organ of Corti. Staining of Hensen's cells and Deiters' cells, especially the outer Deiters' cells, was more intense than staining of the outer hair cells, inner hair cells, and pillar cells. Staining intensity was greatest at the basal turn and progressively decreased in the upper turns with the apex showing the weakest staining pattern. This is the first demonstration of a metabotropic P2Y receptor in the guinea pig organ of Corti. PMID- 14552424 TI - Patients with auditory neuropathy/dys-synchrony lack efferent suppression of transient evoked otoacoustic emissions. AB - Function of the olivocochlear reflex, measured by suppression of transient evoked otoacoustic emissions, is assessed in nine patients with bilateral auditory neuropathy/dys-synchrony and compared to matched control subjects with normal auditory function. TEOAEs were acquired using 65 dB peak sound pressure linear clicks with and without the presence of broad-band noise presented binaurally, ipsilaterally, or contralaterally in a forward masking paradigm. Efferent suppression differed significantly between subject groups (p < .0001). Mean suppression was less than 0.22 dB across all suppressor noise conditions for the auditory neuropathy/dys-synchrony subjects. TEOAE suppression averaged 4.47 (binaural), 2.41 (ipsilateral), and 1.52 (contralateral) dB in the control subjects. Suppression characteristics across the three suppressor conditions were also assessed in one patient with unilateral auditory neuropathy/dys-synchrony. The results suggest that poor efferent responses are related to compromised afferent input to the OCR pathway and support the use of efferent suppression of otoacoustic emissions as a differential measure of auditory function in patients with auditory neuropathy/dys-synchrony. PMID- 14552425 TI - Olivocochlear efferent suppression in classical musicians. AB - Suppression of transient-evoked otoacoustic emissions was recorded from 29 members of the Louisiana Philharmonic Orchestra and 28 non-musician control subjects matched for age and gender. Binaural broad band noise was used as the suppressor stimulus in a forward masking paradigm. Results showed musicians to have significantly more suppression than non-musicians for both the right and left ears. Two possible explanations for this functional difference between groups are that moderately loud music serves as a sound conditioning stimulus and that music can be a mechanism for strengthening central auditory pathways which may influence the olivocochlear reflex arc. Possible explanations for this are discussed and ear, gender, and age differences within each group are examined. Additionally, middle-ear muscle reflex thresholds were found to be higher in musicians than non-musicians at some frequencies in some conditions. PMID- 14552427 TI - Development of auditory asymmetry in transient evoked otoacoustic emissions in pre-term infants. AB - We previously reported that transient evoked otoacoustic emissions (TEOAEs) continue to develop after the onset of cochlear function in pre-term infants from 34 to 39 weeks of conceptional age (Morlet et al., 1996). The time-course of development differed between genders. Reported here are findings of further analysis of data from the study cohort, comprising 1020 ears of 510 pre-term neonates (conceptional age ranging from 34 to 39 weeks), demonstrating developmental differences between right and left ears. The left ear of female pre term infants showed enhancement of TEOAE amplitude at low and medium frequencies with age, whereas differences were less dramatic in the right ear. In male infants, TEOAE amplitude decreased in several frequency bands at high frequencies, above 4 kHz, between 34 and 39 weeks conceptional age; most developmental differences were found to be more dramatic in the right than in the left ear. It is tempting to speculate that these developmental features underlie well-known inter-aural asymmetries that have been demonstrated in the adult human. PMID- 14552426 TI - Active hair bundle movements and the cochlear amplifier. AB - The "active process" is a term used to describe amplification and filtering processes that are essential for obtaining the exquisite sensitivity of hearing organs. Understanding the components of the active process is important both for our understanding of the normal physiology of hearing and because perturbations of the cochlear amplifier may lead to such maladies as threshold shifts (both temporary and permanent), tinnitus, sensorineural hearing loss and presbicusis. To date the cochlear amplifier has largely been attributed to outer hair cell electromotility; however, recent evidence suggests, that active properties of the hair bundle may also be important. Most likely both somatic motility and active hair bundle movements contribute to establishing the cochlear active process. This paper reviews recent evidence regarding known active processes in the hair bundle gating compliance, and fast and slow adaptation. PMID- 14552428 TI - Classification of acute leukemias. AB - Because of the increasing recognition of the importance of genetic events to the diagnosis and treatment of the acute leukemias, the proposed new World Health Organization (WHO) classification incorporates genetic aberrations and immunology as major defining features in addition to morphology. In a hierarchal approach, genetic changes have precedence in the acute myeloid leukemias and immunology and genetic changes have precedence in the acute lymphoblastic leukemias. Four major groups of acute myeloid leukemia are recognized: 1) Acute myeloid leukemias with recurrent genetic abnormalities, 2) Acute myeloid leukemia with multilineage dysplasia, 3) Acute myeloid leukemias, therapy related, and 4) Acute myeloid leukemia not otherwise categorized. Two types of acute lymphoblastic leukemia are recognized based on immunologic characteristics: precursor B lymphoblastic leukemia/lymphoma and precursor T lymphoblastic leukemia/lymphoma. Precursor B acute lymphoblastic leukemia/lymphoma is subclassified into prognostic genetic groups. Biphenotypic leukemia is recognized as a form of acute leukemia of ambiguous lineage. PMID- 14552429 TI - Myelodysplastic syndromes, chronic myeloproliferative diseases, and myelodysplastic/myeloproliferative diseases. AB - This article reviews the major diagnostic criteria for the myelodysplastic syndromes, chronic myeloproliferative diseases, and myelodysplastic/myeloproliferative diseases. Perhaps the most important message this article intends to convey is that the proper diagnosis and classification of myelodysplastic syndromes, chronic myeloproliferative diseases, and myelodysplastic/myeloproliferative diseases requires a multidisciplinary approach that correlates morphologic findings with clinical, genetic, and other laboratory information. Thus, the pathologist is central to the diagnosis of these disorders. Not only do pathologists have the morphologic skills to interpret peripheral blood and bone marrow aspirate smears and bone marrow biopsy specimens properly, but they often are responsible for interpretation of flow-cytometry and molecular genetic data as well. Pathologists are therefore in the best position to determine whether all the individual pieces of data fit together for the diagnosis under consideration. An additional important theme in the paper is that "well-prepared" blood and bone marrow aspirate smears and "adequate, well processsed" bone marrow biopsy specimens are essential for the diagnosis. In the author's opinion, inadequate specimens usually account for most of the difficulties encountered in the proper diagnosis of these diseases. It is hoped that when an excellent specimen is available, the guidelines contained in this article may provide the pathologist with assistance in arriving at the most appropriate diagnosis. PMID- 14552430 TI - Update in the pathologic features of mature B-cell and T/NK-cell leukemias. AB - Modern diagnosis of mature B- and T-cell leukemias requires integration of morphologic, immunophenotypic, and molecular genetic features. This integrative approach has allowed more precise definitions of specific disease entities. This in turn provides better information for clinicians to select proper therapy and determine prognosis. The characteristic pathologic features of these disorders are reviewed. PMID- 14552431 TI - Hodgkin and non-Hodgkin lymphoma involving bone marrow. AB - The role of bone marrow examination in diagnosis and staging of patients with Hodgkin lymphoma, B-non-Hodgkin lymphoma, and T-non-Hodgkin lymphoma is reviewed. Optimal routine and specialized bone marrow examination techniques are discussed. The salient morphologic, immunophenotypic, and genetic features of mature and immature B, T neoplasms and classic Hodgkin lymphoma in bone marrow are delineated, along with recommendations to distinguish these overt neoplasms from non-neoplastic processes. PMID- 14552432 TI - Plasma cell myeloma marrow diagnosis including morphologic and phenotypic features. AB - This article emphasizes both the morphologic and phenotypic features of the bone marrow in plasma cell myeloma. It details the morphologic features of both trephine biopsies and marrow aspirations. It emphasizes the salient phenotypic features of marrow myeloma cells, in contrast with normal plasma cells. The myeloma cell phenotype is discussed from the perspective of both tissue section immunohistochemistry (IHC) and flow cytometry (FACS analysis). The specific criteria for myeloma diagnosis are discussed and illustrated in Figures 1-12. Finally, the emphasis is on the key morphologic and phenotypic diagnostic criteria of each of the plasma cell neoplasms. PMID- 14552433 TI - Histiocytic lesions involving the bone marrow. AB - Histiocytic lesions involving the bone marrow include a number of reactive and neoplastic disorders. This article discusses the morphologic, immunophenotypic, and genotypic features of a variety of diseases associated with histiocytes and/or monocytes. Lysosomal storage disorders and hemophagocytic syndromes are often first diagnosed by bone marrow examination. Granulomas involving the bone marrow may also be the first indication of a systemic disorder. Apart from acute and chronic monocytic leukemias, the bone marrow is rarely involved by malignant histiocytic disorders, of which Langerhans cell histiocytosis is the most common. PMID- 14552434 TI - Bone marrow evaluation in pediatric patients. AB - Pediatric bone marrow evaluation is often challenging, especially for pathologists with more experience evaluating bone marrow specimens from adults. This article reviews the features of several pediatric hematologic malignancies that have been selected because they illustrate the different approach required to evaluate pediatric bone marrow specimens, and highlight potential diagnostic pitfalls. The following topics have been selected for discussion: ancillary studies required for prognostication in pediatric acute lymphoblastic leukemia, the classification of pediatric acute myeloid leukemia, congenital acute leukemia and its distinction from Down syndrome-associated transient myeloproliferative disorder, diagnosis and classification of pediatric myelodysplastic syndromes, and juvenile myelomonocytic leukemia as a distinct disease entity of childhood. PMID- 14552435 TI - The Second Annual Primary Care Conference--Programming to eliminate health disparities among ethnic minority populations: an introduction to proceedings. AB - From October 31, 2002 through November 2, 2002, the Second Annual Primary Care Conference was held, sponsored by the Morehouse School of Medicine's National Center for Primary Care and its Prevention Research Center. The conference was designed as a collaborative activity with the Atlanta Regional Health Forum; The Carter Center; Emory University's School of Medicine, Nell Hodgson Woodruff School of Nursing, and Rollins School of Public Health; Georgia Chapter of the American College of Physicians/American Society of Internal Medicine; Georgia Nurses Foundation; Southeastern Primary Care Consortium, Inc./Atlanta Area Health Education Center; St. Joseph's Mercy Care Services; United States Department of Health and Human Services: Agency for Healthcare Research and Quality; Centers for Disease Control and Prevention; Health and Human Services (Region IV); Health Resources and Services Administration; Office of Minority Health (Region IV); and Office on Women's Health (Region IV). The 2 and a half-day conference featured 5 plenary sessions and 3 tracks of medical education for primary care physicians and other healthcare providers. The tracks were categorized as: Track A: Adult Health; Track B: Public Health and Prevention; and Track C: Maternal/Child/Youth Health. Within each track, 6 working sessions were presented on topic areas including diabetes, obesity, cardiovascular disease, cancer, mental health, infectious disease, behavioral and social health, women's health, stroke, and asthma. A total of 18 working sessions took place and each working session included 3 presentations. Continuing medical education credits or continuing education units were granted to participants. In all, 485 individuals participated in the conference, with the majority of the participants from the southeastern United States. Of the attendees, 35% were physicians (MD); 13% were nurses (RN); 12% held master-level degrees; and 12% held other doctorate-level degrees. PMID- 14552436 TI - Are health disparities on the public health agenda? Where? PMID- 14552437 TI - Beyond rhetoric: what we need to know to eliminate disparities. AB - Racial and ethnic disparities in health care are well documented and can be discussed in 3 broad categories: health system factors, patient-level factors, and patient/provider interaction. Clinicians and others working in health care and related fields are knowledgeable about disparities in health, but the general US population is not. Racial/ethnic disparities are most striking in life expectancy, infant mortality, and lack of health insurance. The inaugural edition of the National Healthcare Disparities Report, due out in 2003, will provide valuable insights into the state of health care in America, including a comprehensive view of disparities in health care. The Agency for Healthcare Research and Quality is conducting and supporting research, data collection, and other initiatives aimed at reducing racial/ethnic disparities in health care. The documented disparities in health care represent a critical opportunity for quality improvement that requires input from all sectors, including policymakers, providers, community leaders, and patients. PMID- 14552438 TI - Public health and primary care: challenges and opportunities for partnerships. PMID- 14552439 TI - HRSA's role in improving community health: creative solutions that work. PMID- 14552440 TI - The Georgia Cancer Coalition. PMID- 14552441 TI - The Atlanta Community Food Bank: strategic lessons for eliminating health disparities. PMID- 14552442 TI - Southeastern Partnership to Eliminate Health Disparities. PMID- 14552443 TI - Taking charge: making partnerships work for better health outcomes. PMID- 14552444 TI - Diabetes, obesity, and the politics of health: What are the real causes of this epidemic? What is its impact? What will it take to turn the tide? PMID- 14552445 TI - Obesity: clinical impact and interventions that work: an update. PMID- 14552446 TI - Diabetes: update on management and treatment. PMID- 14552447 TI - Early diagnosis and treatment in lung cancer: can the primary care provider make a difference? PMID- 14552448 TI - Breast cancer: diagnosis and treatment. PMID- 14552449 TI - Colon cancer: update on treatment. Are outcome disparities real? PMID- 14552450 TI - Heart disease in women. PMID- 14552451 TI - Hormone replacement therapy: where do we go from here? PMID- 14552452 TI - Gynecology in the generalist's office. PMID- 14552453 TI - Update: The Paul Coverdell Georgia Stroke Registry Pilot Prototype. PMID- 14552454 TI - Dementia: new theories, new treatments. PMID- 14552455 TI - Depression: the common denominator--how can we help? PMID- 14552456 TI - Responding to bioterrorism. PMID- 14552457 TI - Strategies for community participation in diabetes prevention: a Detroit experience. AB - The African-American community has been disproportionately burdened by diabetes and its associated complications. Nearly 2.8 million African Americans have diabetes. It is essential to increase community participation in diabetes prevention and health promotion as a method to improve health disparities. To address these issues, the Center for Medical Treatment Programs in Diverse Populations (MedTEP) was developed at Henry Ford Health System. The community participatory strategies described in this paper give a framework, which health systems can use to develop community-based partnerships and improve participation of community members in diabetes prevention and diabetes-related research. The strategies include receiving guidance by community leaders, providing a service, establishing partnerships, and disseminating information to the community. It is the goal of the MedTEP Center to continue to further develop and test models of community outreach to determine the most effective approaches to improve health outcomes and sustain the gain in African-American communities. PMID- 14552458 TI - "Best science" for the reduction of disparities in cancer. PMID- 14552459 TI - Strategies for community participation in cancer prevention. PMID- 14552460 TI - Prostate cancer: to screen or not to screen. PMID- 14552461 TI - A clinician's guide to the medical consequences of smoking and evidence-based strategies for tobacco cessation. PMID- 14552463 TI - Reorganizing health systems to promote best practice medical care, patient self management, and family-centered care for childhood asthma. PMID- 14552462 TI - Mental health issues in disasters and terrorist attacks. AB - Recent events make clear that those living in the United States are at risk of exposure to a variety of potentially traumatic events, ranging from sniper and terrorist attacks to a variety of natural disasters. This paper provides a broad overview of the most common psychological reactions that can be expected in the aftermath of such events, how primary care practitioners can identify such reactions in their patients, and actions those practitioners might take. PMID- 14552464 TI - Middle school internships for at-risk youth: a proven intervention. PMID- 14552466 TI - Family/domestic violence: How to intervene? How to prevent? PMID- 14552467 TI - The role of primary care professionals in nurturing healthy families and healthy communities. PMID- 14552465 TI - Teen sex, teen pregnancy, and teen sexual risky behaviors. PMID- 14552468 TI - Bacterial vaginosis and other maternal infections: treatable causes of the Black White gap? PMID- 14552469 TI - Canada wraps up BSE investigation. PMID- 14552470 TI - Airborne antimicrobials cause concern. PMID- 14552471 TI - Despite high demand, laboratory animal veterinarians in short supply. Economic survey shows upward trend in specialists' salaries. PMID- 14552472 TI - International guests add dimension to convention. PMID- 14552473 TI - Comments on comparison of culling rates among cows. PMID- 14552474 TI - Found thread in unrelated letters. PMID- 14552475 TI - Responses to letter on lack of pain management. PMID- 14552476 TI - Responses to letter on lack of pain management. PMID- 14552477 TI - Feedback continues on Consumer Reports article, AVMA response. PMID- 14552478 TI - Believe epidemiology key to vaccination protocols. PMID- 14552479 TI - What is your diagnosis? Cor triatriatum dexter. PMID- 14552480 TI - Animal behavior case of the month. An English Cocker Spaniel was examined because of growling and snapping at people and dogs. PMID- 14552481 TI - In defense of zoos and aquariums: the ethical basis for keeping wild animals in captivity. PMID- 14552482 TI - Strategic collection planning and individual animal welfare. PMID- 14552483 TI - Positive reinforcement training and environmental enrichment: enhancing animal well-being. PMID- 14552484 TI - Stress and distress: evaluating their impact for the well-being of zoo animals. PMID- 14552485 TI - Steering the ark toward Eden: design for animal well-being. PMID- 14552486 TI - Surplus animals: the price of success. PMID- 14552487 TI - Evaluation of a continuous glucose monitoring system for use in dogs, cats, and horses. AB - OBJECTIVE: To evaluate a continuous glucose monitoring system (CGMS) for use in dogs, cats, and horses. DESIGN: Prospective clinical study. Animals-7 horses, 3 cats, and 4 dogs that were clinically normal and 1 horse, 2 cats, and 3 dogs with diabetes mellitus. PROCEDURE: Interstitial glucose concentrations were monitored and recorded every 5 minutes by use of a CGMS. Interstitial glucose concentrations were compared with whole blood glucose concentrations as determined by a point-of-care glucose meter. Interstitial glucose concentrations were also monitored in 2 clinically normal horses after oral and i.v. administration of glucose. RESULTS: There was a positive correlation between interstitial and whole blood glucose concentrations for clinically normal dogs, cats, and horses and those with diabetes mellitus. Events such as feeding, glucose or insulin administration, restraint, and transport to the clinic were recorded by the owner or clinician and could be identified on the graph and associated with time of occurrence. CONCLUSIONS AND CLINICAL RELEVANCE: Our data indicate that use of CGMS is valid for dogs, cats, and horses. This system alleviated the need for multiple blood samples and the stress associated with obtaining those samples. Because hospitalization was not required, information obtained from the CGMS provided a more accurate assessment of the animal's glucose concentrations for an extended period, compared with measurement of blood glucose concentrations. Use of the CGMS will promote the diagnostic and research potential of serial glucose monitoring. PMID- 14552488 TI - Assessment of plasma concentrations of doxycycline in budgerigars fed medicated seed or water. AB - OBJECTIVE: To determine whether plasma doxycycline concentrations considered effective for treatment of avian chlamydiosis could be safely established and maintained in budgerigars via administration of doxycycline in water or seed. DESIGN: Prospective study. ANIMALS: 68 healthy mature budgerigars. PROCEDURE: In 14-day trials, plasma doxycycline concentrations were measured in budgerigars provided with water containing 0, 50, 100, 200, or 400 mg of doxycycline hyclate/L or a hulled seed diet containing 0, 100, 200, or 400 mg of doxycycline hyclate/kg. On the basis of these results, birds were fed seed containing 300 mg of doxycycline/kg for 42 days, and a control group received unmedicated seed. Blood samples for plasma doxycycline analysis were collected either in the morning on treatment days 4, 7 14, 21, 28, 35, and 42 or in the afternoon on days 12, 26, and 40. Birds were observed daily. On days 14, 28, and 42, physical and plasma biochemical variables, PCV, and total solids concentration were measured; cloacal specimens were obtained for bacteriologic and fungal culture. RESULTS: During a 14-day period, treatment with water containing < or = 400 mg of doxycycline/L did not maintain plasma doxycycline concentrations of > or = 1 microg/mL, but seed containing 300 mg of doxycycline hyclate/kg maintained mean plasma doxycycline concentrations > 0.98 microg/mL for 42 days without notable adverse effects. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that hulled seed containing 300 mg of doxycycline hyclate/kg can safely establish and maintain plasma doxycycline concentrations that are considered adequate for treatment of chlamydiosis in adult nonbreeding budgerigars. PMID- 14552489 TI - Use of a self-expanding occluding stent for nonsurgical closure of patent ductus arteriosus in dogs. AB - OBJECTIVE: To evaluate the clinical application of a catheter-delivered, self expanding occluding stent for closure of patent ductus arteriosus (PDA) in dogs. DESIGN: Prospective study. ANIMALS: 23 client-owned dogs weighing at least 3 kg (6.6 lb). PROCEDURE: Dogs were evaluated by physical examination, electrocardiography, thoracic radiography, and 2-dimensional, M-mode, spectral and color-flow Doppler echocardiography to confirm the diagnosis and obtain baseline measures. Shunt severity and ductal size and anatomy were established by means of angiography. With fluoroscopic guidance, the occluding stent, attached to a delivery cable, was maneuvered though the right side of the heart into the ductus via a prepositioned introducer sheath. After angiographic verification of appropriate stent placement, the delivery cable was detached, and the introducer sheath was withdrawn. Closure of the PDA was evaluated by means of angiography 15 minutes after stent deployment and by echocardiography 1 and 3 months after the procedure. RESULTS: There were no operative deaths. There were 2 deployment failures, both attributable to avoidable operator errors. Angiography performed after stent deployment indicated PDA closure in 13 of 20 (65%) dogs. There were 2 postoperative deaths in dogs with heart failure; both deaths were thought to be unrelated to use of the occluding stent. Complete PDA closure, determined by Doppler color-flow echocardiography, was evident in 17 of 19 dogs within 3 months and in 1 additional dog within 1 year of stent deployment, resulting in closure in 18 of 19 dogs completing the study protocol. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that a catheter-delivered occluding stent can be used successfully to close PDAs in dogs. PMID- 14552490 TI - Effects of postoperative administration of ketoprofen or carprofen on short- and long-term results of femoral head and neck excision in dogs. AB - OBJECTIVE: To determine whether postoperative administration of ketoprofen or carprofen had any effects on short- or long-term results of femoral head and neck excision (FHNE) in dogs. DESIGN: Prospective randomized controlled trial. ANIMALS: 40 client-owned, large-breed dogs undergoing FHNE and 15 healthy large breed dogs used as controls for hip joint angle measurements and force plate analyses. PROCEDURE: Dogs undergoing FHNE were treated with ketoprofen, carprofen, or a placebo for 21 days after surgery. Hip joint abduction and extension angles were measured at the end of surgery and 120 days later. Lameness scores were assigned, and force plate analyses were performed on days 3, 15, and 120. RESULTS: There were no significant differences among treatment groups in regard to hip joint angles or lameness scores. Force plate analysis revealed that dogs in all 3 treatment groups bore consistently less weight on the operated limb than did control dogs for the duration of the study. Dogs receiving ketoprofen had greater peak propulsive force at a walk on day 3 and greater peak vertical force at a walk on day 15 than did dogs receiving the placebo. Treatment of an acute condition and preservation of the lesser trochanter, but not postoperative analgesic administration, were positively associated with ground reaction forces on day 120. Owners of 12 of 31 dogs indicated that the dog's gait worsened for a few days after discontinuation of analgesic administration. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of ketoprofen or carprofen after surgery was not associated with long-term results of FHNE, probably because of the impact of other factors. Because some owners noticed worsening of the lameness following cessation of analgesic administration in the present study, it is possible that longer administration would have improved long-term results. PMID- 14552491 TI - Psoriasiform-lichenoid-like dermatosis in three dogs treated with microemulsified cyclosporine A. AB - Cyclosporine has been reported to be effective for the treatment of various cutaneous autoimmune disorders in dogs. Adverse reactions have generally been limited to gastrointestinal tract disturbances and cutaneous eruptions. The article describes antimicrobial-responsive cutaneous reactions in 3 dogs being treated with microemulsified cyclosporine A because of various dermatologic conditions. Cutaneous reactions in these dogs were similar to psoriasiform lichenoid dermatitis and may represent an atypical staphylococcal infection. PMID- 14552492 TI - Multicentric leiomyosarcoma in a young male horse. AB - A 2-year-old male Thoroughbred was evaluated for multiple firm, raised nodules of the maxilla and mandible that had been present since 6 months of age. The horse was evaluated as a yearling with a final histologic diagnosis of fibrosarcoma. As a 2-year-old, the colt made a loud upper airway sound and was agitated after strenuous exercise. Results of upper airway endoscopy indicated severe narrowing of the nasal passages and pharynx with normal resting function. Radiography revealed multiple circular bony lesions in the mandible, maxilla, multiple long bones, and cervical vertebrae. Biopsy specimens of the left mandible and right facial bone were obtained. A diagnosis of leiomyosarcoma with extensive metastatic bone formation was made on the basis of histologic features and the immunohistochemical staining properties of the biopsy specimens. Because of the diffuse distribution of the lesions, prognosis was guarded; 4 years after diagnosis, the horse had not pursued an athletic career. PMID- 14552493 TI - Evaluation of bacteriologic culture of individual and pooled fecal samples for detection of Mycobacterium paratuberculosis in dairy cattle herds. AB - OBJECTIVES: To determine the sensitivity of bacteriologic culture of pooled fecal samples in detecting Mycobacterium paratuberculosis, compared with bacteriologic culture of individual fecal samples in dairy cattle herds. STUDY DESIGN: Cross sectional study. ANIMALS: 24 dairy cattle herds. PROCEDURE: Individual and pooled fecal samples were submitted for bacteriologic culture, and results were compared between these groups. RESULTS: Ninety-four and 88% of pooled fecal samples that contained feces from at least 1 animal with high (mean, > or = 50 colonies/tube) and moderate (mean, 10 to 49 colonies/tube) concentrations of M paratuberculosis, respectively, were identified by use of bacteriologic culture of pooled fecal samples. Prevalences of paratuberculosis determined by bacteriologic culture of pooled and individual fecal samples were highly correlated. CONCLUSIONS AND CLINICAL RELEVANCE: Bacteriologic culture of pooled fecal samples provided a valid and cost-effective method for the detection of M paratuberculosis infection in dairy cattle herds and can be used to estimate prevalence of infection within a herd. PMID- 14552494 TI - Babesia odocoilei infection in elk. AB - Two male North American elk from a commercial herd were evaluated because of a sudden onset of lethargy, anorexia, and voiding of red urine. These 2 elk were kept in the same pen as 4 other male elk that had died during the preceding 2 months. Laboratory analyses revealed anemia and intraerythrocytic parasites, later confirmed as Babesia odocoilei (a protozoal hemoparasite of cervids). Of the 240 elk remaining in the herd, 59 were screened for B odocoilei by microscopic evaluation of blood smears, protozoal culture of blood, and immunofluorescent antibody testing of serum. Of those 59 elk, 34 (58%) were infected with B odocoilei. Babesia odocoilei infection in elk can be fatal and should be considered in cases of sudden death or acute hemolytic anemia. Familiarity with the disease in elk is essential for practitioners because of the increasing popularity of commercial elk farming. PMID- 14552495 TI - Racial disparity in the pharmacological management of schizophrenia. AB - This study investigated racial differences in the prescription of psychopharmacologic treatments to individuals with schizophrenia. Data were derived from a patient survey and medical record review for 344 persons with schizophrenia recruited from outpatient psychiatric facilities in two States in the Schizophrenia Patient Outcomes Research Team study. African-Americans were three times more likely to receive depot antipsychotic medications (odds ratio [OR]: 2.91; 95% confidence interval [CI]: 1.68-5.01) and 76 percent less likely to receive new-generation antipsychotic medications (OR: 0.24; 95% CI: 0.12 0.46), compared to their Caucasian counterparts. Chlorpromazine-equivalent antipsychotic dosages did not differ significantly between African-American and Caucasian patients. Compared to Caucasians, a larger proportion of African Americans received antiparkinsonian medications (63% vs. 48%, chi2 = 7.01; df = 1; p = 0.008), but African-Americans were less than half as likely to receive adjunctive psychopharmacologic treatments (OR: 0.43; 95% CI: 0.27-0.71). Pronounced racial variations in the psychopharmacologic management of schizophrenia in typical clinical practice settings were observed and persisted when analyses were adjusted for selected patient demographic and clinical characteristics. A prospective, longitudinal evaluation is warranted to determine whether the observed patterns of prescribing are associated with poorer therapeutic outcomes in minority patients. PMID- 14552496 TI - Who are patients on conventional antipsychotics? AB - The advent of atypical antipsychotics presented psychiatrists with an effective way of treating psychosis without the many side effects associated with conventional agents. Given the superiority of atypical antipsychotics, we examined demographic information and treatment histories of patients currently treated with conventional antipsychotics, especially in regard to treatment with atypical agents. Medication histories and demographic information for 276 patients admitted to an urban hospital were obtained by patient/family interviews and review of medical records. Chi-square and logistic regression tests were used to analyze data for possible predictive factors of which patients within the sample were still receiving conventional antipsychotics for treatment. Seventy eight (28%) patients were currently being treated with conventional antipsychotics. More than half of them had never received a trial of an atypical agent. African-Americans, who are more likely to suffer adverse effects from conventional antipsychotics, and substance abusers were overrepresented in this group. It is unclear to what extent ethnic or cultural bias played a role in determining medication choice. Because conventional antipsychotics are associated with more side effects and greater medication nonadherence, these patients should be evaluated for appropriateness of a trial with an atypical agent even if they are currently stable with a conventional antipsychotic. PMID- 14552497 TI - Met and unmet needs of schizophrenia patients in a Spanish sample. AB - Deinstitutionalization of people with schizophrenia increases the importance of evaluating their needs. This study set out to identify the most common needs of people with schizophrenia who live in the community, analyze how those needs differ when evaluated by staff or by patients, describe the kind of help patients receive, and find out the variables that correlate with having unmet needs. A random sample of 231 outpatients with schizophrenia were evaluated with the Camberwell Assessment of Need and other predictor and outcome variables. Staff detected more needs than patients did. Mean number of needs as rated by patients was 5.36 and staff 6.6 (p < 0.001). Mean number of unmet needs was also greater when assessed by staff than by patients: 1.38 versus 1.82 (p < 0.001). The most frequently detected needs by patients involved psychotic symptoms, house upkeep, food, and information. Staff most often detected needs involving psychotic symptoms, company, daytime activities, house upkeep, food, and information. In a multiple regression model, needs were weakly associated with the clinical variables and quality of life. Needs assessment is complementary to clinical evaluation in schizophrenia. PMID- 14552498 TI - Disease management in Latinos with schizophrenia: a family-assisted, skills training approach. AB - This study evaluated the effectiveness of a skills training program designed to teach disease management to Latinos with schizophrenia treated at a community mental health center. Ninety-two Latino outpatients with schizophrenia and their designated relatives were randomly assigned to 3 months of skills training (ST) versus customary outpatient care (CC) and followed for a total of 9 months. The skills training approach was culturally adapted mainly by including the active participation of key relatives to facilitate acquisition and generalization of disease management skills into the patients' natural environment. There was a significant advantage for the ST group over the CC group on several symptom measures, skill acquisition and generalization, level of functioning, and rates of rehospitalization. There were no significant differences between the groups on quality of life or caregiver burden. Skills training had a direct effect on skill acquisition and generalization, and utilization of disease management skills led to decreased rates of rehospitalization. Incorporating an intensive, culturally relevant generalization effort into skills training for Latinos with schizophrenia appeared to be effective in teaching disease management and viable in a community mental health center. PMID- 14552499 TI - Satisfaction with mental health services among people with schizophrenia in five European sites: results from the EPSILON Study. AB - Patient satisfaction with services is an important outcome variable that is increasingly used in mental health service evaluation. This study includes 404 people with schizophrenia in five European sites and addresses five questions focused on site, service, and patient characteristics as variables that might explain service satisfaction, using the Verona Service Satisfaction Scale. Patient satisfaction differed significantly across sites (highest in Copenhagen, lowest in London). In all sites, patients were least satisfied with involvement of relatives in care and information about illness. A multiple regression model showed that lower levels of total service satisfaction were associated with living in London or Santander, being retired/unemployed, having more hospital admissions, having more severe psychopathology, having more unmet needs, or having lower satisfaction with life. This model explained 31 percent of variance in service satisfaction. Our data show that service satisfaction can be seen as a result of (1) the ability of the service to provide a standard of care above a certain quality threshold, and (2) the perception of each patient that the care received has been tailored to the patient's own problems. PMID- 14552500 TI - The Schizophrenia Care and Assessment Program Health Questionnaire (SCAP-HQ): an instrument to assess outcomes of schizophrenia care. AB - Advances in treatment technologies and development of evidence-based standards of care demand better methods for routine assessment of outcomes for schizophrenia in systems of care. This article describes the development and psychometrics of a new instrument to assess outcomes of routine care for persons with schizophrenia in service systems. Candidate items for the Schizophrenia Care and Assessment Program Health Questionnaire (SCAP-HQ) were drawn from existing measures. Domains covered include disease outcomes (symptoms, subjective medication effects, substance abuse), functional status, health status, quality of life, and public safety. A sample of 1,584 patients with schizophrenia or schizoaffective disorder who were recruited into a large prospective, naturalistic study on the course of treatment for schizophrenia completed the SCAP-HQ at baseline and 1 year later (n = 434), providing data for factor analysis, assessment of internal consistency, convergent validity, and responsiveness to change. A subsample of 121 patients completed a test-retest protocol. Fifteen scales were derived by factor analysis from 55 outcome items on the SCAP-HQ. These factors covered psychiatric symptoms, life satisfaction, instrumental activities of daily living, health-related disability, subjective medication side effects, vitality, legal problems, social relations, mental health-related disability, suicidality, drug and alcohol use, daily activities, victimization, violence, and employment. For most scales, standard psychometric parameters, including internal consistency and test-retest reliability, convergent validity, and responsiveness to change, were acceptable for application to large sample evaluations of care systems. This new measure represents an advance in the development of outcome measures for schizophrenia for use in large-scale studies of routine care. PMID- 14552501 TI - Preference weights for cost-outcome analyses of schizophrenia treatments: comparison of four stakeholder groups. AB - This study quantified preferences for schizophrenia outcomes in four stakeholder groups, tested the hypotheses that outcomes differ in importance and stakeholder groups have different preferences, and produced preference weights for seven outcomes for cost-outcome analysis. Fifty patients with schizophrenia, 50 clinicians, 41 family members of patients, and 50 members of the general public rated 16 schizophrenia-related health states, yielding preference weights for seven outcomes: positive symptoms, negative symptoms, extrapyramidal symptoms, tardive dyskinesia, social function, independent living, and vocational function. Outcomes differed in importance (F = 23.4, p < 0.01). All stakeholders rated positive symptoms and social functioning as more important than negative and extrapyramidal symptoms. Stakeholder groups had different preferences (F = 1.9, p = 0.01). Patients rated extrapyramidal symptoms as more important than did other groups (p < 0.01); clinicians rated social functioning as more important than did patients or family members (p < 0.05); and clinicians and family members rated vocational functioning as more important than did patients and the general public (p < 0.05). Results show that schizophrenia outcomes are not equally important and that stakeholder groups value outcomes differently, demonstrating the importance of incorporating stakeholder preferences in cost-outcome analyses and other treatment comparisons. PMID- 14552502 TI - Risk adjustment in studies using administrative data. AB - Administrative data are increasingly used to evaluate service use and cost of treatments in "real world" settings. However, the degree to which administrative data can be used to risk-adjust for differences between nonrandomized groups receiving different treatments has not been studied. This study used data from a large randomized trial to determine whether administrative data describing previous utilization and cost are as good as clinical data in predicting future resource utilization and cost. Clinical data (symptoms, quality of life, pharmacological side effects) and administrative data (inpatient utilization and total cost) were obtained from a large randomized clinical trial comparing clozapine and haloperidol. The combination of administrative measures prior to randomization was the best predictor in a multiple regression model of service use in the year after randomization (adjusted r2 = 0.27-0.31). Models using administrative data alone predicted 24 to 28 percent of the variance of service use and costs, while clinical data predicted only 6 to 7 percent of the variation in the same dependent variables. In this analysis, administrative data about previous utilization and costs were far more successful than clinical data alone at predicting future utilization and costs and are therefore likely to be better risk adjusters in studies with nonrandomized comparison groups. PMID- 14552503 TI - Schizotypal personality traits in nonpsychotic relatives are associated with positive symptoms in psychotic probands. AB - There remains disagreement over whether increased risk of schizotypal personality disorder (SPD) is confined to the relatives of patients with a diagnosis of schizophrenia or whether it is a more general characteristic of the relatives of all psychotic patients. To examine the relationship between schizotypal dimensions in relatives and psychopathological syndromes in patients with functional psychoses, factor analysis was carried out on (1) ratings from Present State Examination (PSE) interviews with 172 consecutively admitted patients with psychosis (52% of them with schizophrenia), and (2) ratings on items from three schizotypal scales concerning 263 of their nonpsychotic first degree relatives. The factors derived from the patients' PSE interviews were correlated with the schizotypal factors and the nine DSM-IV criteria for SPD concerning the relatives and subjected to a canonical correlation analysis. In this study, no differences were observed concerning the distribution of schizotypal factors or DSM-IV schizotypal features in the relatives of patients with different psychotic diagnoses. However, a syndrome characterized by delusions, hallucinations, and thought interference (positive symptoms) in patients was correlated with high scores on the three schizotypy scales and with positive and negative schizotypal features in relatives. PMID- 14552504 TI - Neuromotor functioning in adolescents with schizotypal personality disorder: associations with symptoms and neurocognition. AB - Previously, Neumann and Walker (1999) used a computerized motor assessment and found that adults with schizotypal personality disorder (SPD) displayed increased and more variable motor force compared to adults with other personality disorders or healthy controls. Using the same motor assessment, the current study examined whether an independent sample of adolescents with SPD manifested a similar pattern of motor dysfunction compared to adolescents with other Axis II disorders or those without a disorder. As predicted, the SPD group showed increased and more variable force compared to the other two groups, which did not differ on these measures. These same motor variables were correlated with negative symptoms, as well as perseverative responses on a card sorting test. The significant correlations between motor functioning and perseverative responses and negative symptoms support recent research suggesting that subcortical motor regions play a role in higher order cognition and negative symptoms. Differences as well as broad similarities in the pattern of motor findings between the adult and adolescent SPD studies are discussed. PMID- 14552505 TI - Using neurophysiological markers of genetic risk to define the boundaries of the schizophrenia spectrum phenotype. AB - There is considerable evidence that schizophrenia spectrum personality (SSP) disorders mark genetic risk for schizophrenia. Use of the spectrum phenotype in genetic and neurophysiological studies may prove informative. However, the degree to which the current diagnostic criteria correspond with genetic risk is unclear. This can be assessed by observing how measures of liability among SSP subjects change as a function of diagnostic criteria. In this study the generalized estimating equation method was used to assess changes in eye-tracking performance among SSP and non-SSP family and community groups employing various diagnostic criteria. Eye-tracking deficits among SSP relatives remained statistically higher compared with the other groups across progressively more liberal SSP criteria. The results suggest that fewer traits than are used in clinical diagnoses can effectively identify the spectrum phenotype among relatives of schizophrenia patients. Thus, reduced criteria may be used in research to increase "high risk" sample size and power to detect neurophysiological and genetic differences. Our results provide suggestive evidence that the use of clinical criteria in research may, in fact, underidentify at-risk individuals--potentially distorting genetic and neurophysiological findings. PMID- 14552506 TI - What is schizophrenic in acute and transient psychotic disorder? AB - Acute and transient psychotic disorder (ATPD) is supposed to differ from schizophrenia, but little research has been done on the subject. In a prospective longitudinal case control study we compared all inpatients with ATPD (ICD-10 F23) treated at Halle University Hospital during a 5-year period with matched controls with "positive" schizophrenia (PS) and with mentally healthy controls. Followup investigations were performed at a mean of 2.2 years after the index episode or 8.2 years after the first episode. Female preponderance in ATPD was marked (78.6%). ATPD and PS patients were similar to each other (but different from healthy controls) in the prevalence of a "broken home" situation and a family history for mental disorders. Compared with PS patients, ATPD patients showed better premorbid social adaptation, and they more often displayed rapidly changing symptoms in the index episode and a negative life event preceding the episode. Despite comparable relapse rates, at followup ATPD patients showed better social adaptation, less psychological impairment, and better global functioning than PS patients. These data support the delineation of ATPD from schizophrenia. PMID- 14552507 TI - Modeling the early course of schizophrenia. AB - Using the Interview for the Retrospective Assessment of the Onset of Schizophrenia (IRAOS), we assessed 170 first illness episodes with a nonpsychotic prodromal stage (73% of the population-based Age, Beginning, Course [ABC] study sample of 232 first illness episodes of schizophrenia from a German population of about 1.5 million). Conrad's (1958) and Docherty et al.'s (1978) stage models of the early course presume unidirectional and compelling patterns of symptom manifestation. Using structural equation modeling, we tested the explanatory power of the stages as latent variables and to what extent these models tally with each other and with data on symptom onset. The models neither converged nor were they confirmed. The reasons for and possible implications of this result will be discussed. We also tested, using various techniques, a causal model of the determinants of social course. The only significant predictors of 5-year social outcome turned out to be social development at psychosis onset and the socially adverse illness behavior of young men. The influence of the traditional predictors, age and gender, type of onset (chronic, acute), and symptomatology, was mediated by these two variables assessed at the end of the prodromal stage. PMID- 14552508 TI - Duration of untreated psychosis and outcome of schizophrenia: delay in intensive psychosocial treatment versus delay in treatment with antipsychotic medication. AB - Duration of untreated psychosis (DUP), defined as delay in treatment with antipsychotic medication, was found to be associated with an unfavorable course of schizophrenia. Delay in intensive psychosocial treatment (DIPT) may also be related to outcome. We examined the relationship of DUP and DIPT with several outcome domains 6 years after onset in a cohort of 88 consecutively admitted patients with early-onset schizophrenia and related disorders. Patients and their parents completed an inventory concerning DUP, DIPT, and various aspects of outcome. Psychotic relapse during the first year after hospitalization was assessed with a chart review. Both DUP and DIPT were found to be associated with negative symptoms at outcome; mode of onset was not. DUP was associated with mild psychotic relapse. DIPT was associated with months of rehospitalization. There was no relation between DUP or DIPT and other aspects of outcome. When we controlled for age at onset, gender, and duration of treated first psychotic episode, only DIPT was associated with negative symptoms at outcome. DIPT may be a more important predictor of negative symptoms at outcome than is delay in starting antipsychotic medication alone. PMID- 14552509 TI - When does experience of psychosis result in a need for care? A prospective general population study. AB - Not all individuals with experience of psychosis develop a need for care. The present study investigated differences in coping strategies and associated levels of perceived control over psychotic symptoms, in relation to need for care status. The influence of coping on need for care was assessed in 47 individuals incident for psychosis in a general population sample of 4,722. Need for care was associated with severity of psychotic experiences rather than associated distress, mean level of control, or average number of coping strategies used. Qualitative differences were apparent, however, in that those who resorted to the strategy of symptomatic coping (a coping strategy characterized by going along with and indulging in symptoms) experienced less control over their symptoms (odds ratio [OR] = 0.79, 95% confidence interval [CI]: 0.63-0.98) and had a higher probability of need for care (OR = 6.07, 95% CI: 1.94-18.95). The results suggest that qualitative differences in self-initiated coping modify the risk for need for care and subsequent patient status in those who experience psychotic symptoms and point to the possible importance of early coping-based interventions. PMID- 14552510 TI - A review of cognitive training in schizophrenia. AB - Empirically supported treatments for schizophrenia now include a variety of psychosocial interventions, such as social skills training, vocational rehabilitation, and psychotherapy. As awareness of the functional importance of neurocognitive impairments in schizophrenia has increased, interest in treatments to improve cognition has grown. We review the literature on cognitive training (CT), which has been studied in 17 published randomized, controlled trials to date. The differential effectiveness of noncomputerized and computer-assisted interventions, with and without strategy coaching, and an environmental adaptation intervention, is examined. We conclude that the different types of approaches, whether computer assisted or not, all have effective components that hold promise for improving cognitive performance, symptoms, and everyday functioning. Our recommendations for further research, including the use of functional outcome measures and long-term followup, highlight the importance of improving ecological validity in this area of treatment research. PMID- 14552511 TI - The effects of a documentary film about schizophrenia on psychiatric stigma. AB - This study examined whether viewing a documentary that depicts individuals with schizophrenia can reduce psychiatric stigma. One hundred and sixty-three individuals were randomly assigned to one of four conditions: no documentary film, documentary about polar bears, documentary about fears of being overweight, and documentary about schizophrenia. Participants also completed a battery of tasks assessing attitudes toward persons with schizophrenia, attributions about the disorder, and intentions to interact with individuals with schizophrenia. The findings showed that compared to the other experimental conditions, the documentary about schizophrenia resulted in more benign attributions about schizophrenia (e.g., less likely to blame individuals with schizophrenia for the disorder) but did not change general attitudes about schizophrenia (e.g., perceived dangerousness). The film also did not increase participants' intentions to interact with persons with schizophrenia. These findings could not be attributed to mood changes associated with the film or how much participants liked the film. The findings provide partial support for the hypothesis that a media depiction of persons with schizophrenia can reduce stigma. PMID- 14552513 TI - Home sweet home. PMID- 14552512 TI - Nutritional assessment of patients with schizophrenia: a preliminary study. AB - The prevalence of obesity in the United States population is increasing, and similar trends can be observed among schizophrenia patients. No thorough examination of the actual nutritional composition of the diet of schizophrenia patients in the United States has been carried out. We therefore employed a 24 hour diet recall in 146 schizophrenia outpatients to gather information on different nutritional variables, such as total caloric intake and total fat, protein, carbohydrate, cholesterol, and fiber content. Data were subsequently compared to data for the general population collected in the Third National Health and Nutrition Examination Survey (NHANES III). Schizophrenia patients as a group ate more food when compared to NHANES III subjects, but the relative percentages of calories derived from fat, protein, and carbohydrates were not found to be different. Therefore, it is unlikely that schizophrenia patients make dietary choices different from those of people in the general population. Instead, schizophrenia patients seem to eat more of the same food. PMID- 14552515 TI - Post-infectious acute cerebellar ataxia in children. AB - Acute cerebellar ataxia is a relatively common neurologic disorder among children. Our aim was to characterize the clinical picture, etiology, and prognosis of acute cerebellar ataxia. The medical records of all children with a diagnosis of acute cerebellar ataxia hospitalized in our center and Hasharon Medical Center from 1990 to 2001 were reviewed. The diagnosis of acute cerebellar ataxia was based on the following criteria: acute onset of ataxia with or without nystagmus; absence of known genetic predisposing factors, such as familial degenerative disorders; and absence of drug intoxication, bacterial meningitis, and metabolic disorders. Thirty-nine children were identified; 54% were male; mean age at presentation was 4.8 +/- 3.8 years. All patients were observed for at least 1 year. A prodromal febrile illness was noted in 74.4%: varicella, 31%; mumps, 20%; nonspecific viral infection, 15.4%; mycoplasma, 5%; Epstein Barr virus, 3%. Latency from the prodromal illness to the onset of ataxia was 8.8 +/- 7.4 days. The most common associated neurologic findings were nystagmus and dysmetria. Full gait recovery took less than 2 weeks on average, and the longest duration of neurologic signs was 24 days (mumps-related). Acute cerebellar ataxia in childhood is a self-limited disease. The recovery was faster than that reported in previous publications and was complete in all children without any neurologic sequelae. Imaging studies are needed only in atypical presentation or if there is no spontaneous improvement after 1 to 2 weeks. PMID- 14552514 TI - The female athlete triad no pain, no gain? PMID- 14552516 TI - The predictive accuracy of pre-adoption video review in adoptees from Russian and Eastern European orphanages. AB - Many internationally adopted children have a pre-adoption video for prospective adoptive parents to review before their commitment to adopt. No published report to date has examined the value of the pre-adoption video evaluation (PreAVE) as a predictor of post-adoption developmental status. The present study was designed to determine whether PreAVE can predict the post-adoption developmental status of internationally adopted children. In this retrospective chart review, children who presented to the Yale International Adoption Clinic between December 1998 and September 2000, and had both a PreAVE and a post-adoption developmental evaluation (PosADE) were selected (N = 20) PreAVE was done using the Denver II and PosADE was done using the Bayley Scales of Infant Development, 2nd edition. The Pearson r coefficient between the two ratings was determined and sensitivity and specificity were calculated. There was a significant correlation between the PreAVE and the PosADEs (r = 0.53, p = 0.01).The sensitivity of PreAVE to detect moderate to severe developmental delay was 43% and the specificity was 85%. Although ratings of development on PreAVE are similar to PosADE, the ability to detect moderate to severe developmental delay by video review is limited. PMID- 14552517 TI - Identifying risk factors for the development of diabetic ketoacidosis in new onset type 1 diabetes mellitus. AB - To identify risk factors associated with the development of diabetic ketoacidosis (DKA) in new-onset type 1 diabetes mellitus, a retrospective chart review of 139 new onset type 1 diabetes patients from 1995 to 1998 was conducted. Categorical data were examined with contingency table analysis. Age range was 0.5 to 18 years. Overall, 38% of the patients presented in DKA. Sixty-two percent of the patients with either Medicaid or no insurance presented in DKA compared to 34% of the patients with private insurance, odds ratio 3.17 (92% CI 1.2-8.3) p = 0.03. Sixty-eight percent of patients in whom the diagnosis was missed (n = 25) presented in DKA (mean age, 5.4 +/- 4.4 years) compared to 32% in whom the diagnosis was not missed (mean age, 8.8 +/- 4.0 years) odds ratio 4.6 (95% CI 1.9 11.7), p = 0.0012; age p = 0.00019. Lack of private insurance, although a risk factor for the development of DKA, did not increase the likelihood of a missed diagnosis. Lack of private insurance (a proxy for socioeconomic status) and young age are apparent risk factors for the development of ketoacidosis. Misdiagnosis by the physician at initial patient encounter is especially prevalent in the young child but not related to insurance. Both increased public awareness and greater medical alertness are necessary to reduce the high rates of DKA in new onset type 1 diabetic children. PMID- 14552518 TI - Screening for domestic violence in the pediatric office: a multipractice experience. AB - The purpose of our study was to screen for domestic violence (DV) in 4 different pediatric practice settings. Women who accompanied their children to well-child visits were eligible. The women were screened with a 6-question tool previously piloted by our group, which included questions on partner abuse, child abuse, and pet abuse. Over a 1-year period, 435 women were screened. Of these women, 95 (22%) described DV at some point in their lives. Sixty-nine (16%) reported abuse longer than 2 years before the screen and 26 (6%) reported more recent abuse. While 11 of 117 women screened in the more affluent private practice reported a history of past abuse, no women in that group reported DV occurring within 24 months. The proportion of women reporting violence did not differ significantly by site, but the proportion of patients reporting new violence was significantly lower at the private practice site by Chi-square analysis. In conclusion, women screened in a variety of pediatric settings will disclose DV. Recent abuse is more likely to be reported in settings with indigent patients. All pediatricians should be screening for DV and have protocols in place to offer women the services they need if DV is revealed. PMID- 14552519 TI - Improvements in immunization compliance using a computerized tracking system for inner city clinics. AB - Vaccination compliance rates were calculated for 1995 to 2001 for enrolled patients, based on the Centers for Disease Control and Prevention guidelines and age-appropriate vaccine schedules. The results reported here indicate computerized tracking with the Doctor's Pediatric Immunization Program (Dr. PIP) maintained vaccine compliance rates (> 90%) in healthy and immunocompromised children at 2 months and 12 months of age. Instituting the computerized system has yielded nearly optimal results in both indigenous inner-city clinics. Despite the efficient progress made by automated tracking, the results for specific vaccine strategies (Varicella) and target groups (human immunodeficiency virus, high-risk indigent populations) may require on-going and intensive educational efforts to achieve optimization levels. PMID- 14552520 TI - Parental compliance with after hours telephone triage advice: nurse advice service versus on-call pediatricians. AB - To compare parental compliance with after-hours triage advice provided by telephone advice nurses and on-call pediatricians, a randomized controlled trial was undertaken at a university general pediatrics practice that enrolled parents or guardians calling for after-hours advice. Advice calls were randomized to a call center advice nurse or the on-call pediatrician. Parental compliance with the triage advice and agreement of the parental report of advice with the pediatrician/nurse report of advice given was evaluated. There were 566 participants in the pediatrician and 616 in the nurse group. Compliance with advice (pediatrician v. nurse) was not significantly different for emergent/urgent care (75.8% v. 72.6%) and self care (74.3% v. 77.2%) but was significantly higher in the pediatrician group for office care (51.5% v. 29.6%; 95% CI of difference, 8.9%-34.2%). Overall agreement between the caller reported and physician or nurse advice was 84.5% for emergent/ urgent, 42.7% for office care, and 93.7% for self-care. PMID- 14552521 TI - The impact of solid-phase gastric emptying studies in the management of children with dyspepsia. AB - Information on the utility of solid-phase gastric emptying studies (SPGES) in the evaluation of children with symptoms of upper gastrointestinal (GI) motor dysfunction is limited. This study was conducted to evaluate the impact of SPGES in the clinical management and outcome of children with upper GI symptoms suggestive of gastroparesis. The records of 45 children who underwent SPGES (31F; 3-17 years) were reviewed. All patients had GI symptoms suggesting gastroparesis. Patients were fed with Tc-99m-sulfur colloid-labeled chicken liver. Adult normal half-life (T1/2) values (F 103 +/- 14 minutes; M 66 +/- 13.6 minutes) were used. The relationships among symptoms, treatment, and outcome were evaluated. Of the 45 patients 9 had delayed, 16 had rapid, and 20 had normal gastric emptying. Six of 9 patients with delayed gastric emptying responded to cisapride. Four of 16 patients with rapid emptying were diagnosed with the dumping syndrome. Of the children with rapid gastric emptying, 87% were females. Twenty patients with normal emptying were diagnosed with gastroesophageal reflux (8), nonulcer dyspepsia (5), irritable bowel syndrome (2), Helicobacter pylori (1), lactose intolerance (1), eosinophilic gastroenteritis (1), duodenitis (1), and constipation (1). In patients who had SPGES for possible gastroparesis, 20% had gastroparesis, 36% had rapid gastric emptying, and 44% had normal gastric emptying. The high number of females in the rapid gastric emptying group might be secondary to normal adult female T1/2 values that were used. The practice of using adult normal T1/2 values in prepubertal girls may need to be revised. Patients with delayed gastric emptying responded to cisapride. PMID- 14552522 TI - Preventive services: blood pressure checks at well child visits. AB - The objective of this study was to determine rates of blood pressure (BP) screening at well-child visits as recommended by the Task Force on Blood Pressure Control in Children. The 1985 and 1996 National Ambulatory Medical Care Surveys were analyzed for changes in proportion of well visits for children aged 3-18 years at which BP was checked. Patient and physician demographics are described. BP screening increased from 50% in 1985 to 61% in 1996. For pediatricians, the estimates were 50% (95% CI, 43-57) and 60% (95% CI, 53-68). For family/general medicine the estimates were 51% (95% CI, 34-69) and 58% (95% CI, 43-74). Age, geographic location, and length of a visit were significant in predicting BP screening. Gender, race, ethnicity, or urban location were not. A stepwise logistic regression confirmed these results. Rates of screening BP at well-child visits have increased but fall short of current recommendations. High-risk children are not screened at a rate different from their lower risk peers. PMID- 14552523 TI - A look at the pediatrician as parent: experiences with the introduction of varicella vaccine. AB - Many physicians voiced reservations about routine use of the varicella vaccine for healthy children after its licensing in 1995. Anecdotal evidence suggested that some pediatricians who were parents themselves were electing not to vaccinate their own children against chickenpox. Little has been written about pediatrician-parents' behaviors in caring for their own children, and how these practices may differ from the behavior that these same pediatricians apply in practice. Pediatricians' tacit attitudes toward medical interventions might be better understood from their behaviors as parents than from their clinical practices, which may be influenced by a sense of responsibility to follow guidelines of professional organizations. Varicella vaccination practices were examined to determine whether pediatricians' behaviors in parenting their own children differed from their recommendations for their patients. A mail survey was sent to 1,762 New York State pediatricians selected randomly from the membership directory of the American Academy of Pediatrics. The response rate was 43% (764/1,762); 63% of responding pediatricians were parents. Eighty-five percent of pediatricians recommended varicella vaccine routinely in practice. Of the pediatricians' own eligible children, 88% (256/291) had been vaccinated against chickenpox. We found a high overall rate of compliance with recommendations for routine use of varicella vaccine. Pediatricians who were parents were just as likely as nonparent pediatricians to recommend the vaccine routinely for their patients. Importantly, pediatrician-parents demonstrated no "double standard"; at the time of the study, 88% of pediatricians' own eligible children had been vaccinated against varicella. PMID- 14552524 TI - Hypoglycemia complicating dehydration due to acute gastroenteritis. AB - To identify clinical variables predictive of hypoglycemia in dehydrated children with acute gastroenteritis (AGE), clinical variables were recorded for dehydrated children younger than 5 years old with AGE before measuring serum glucose. One hundred and eighty-four children were enrolled; 62 with, and 122 without, hypoglycemia. Multivariate analysis identified variables associated with hypoglycemia. Female gender, neurologic symptoms of hypoglycemia, and a greater amount of vomiting vs. diarrhea were significantly associated with hypoglycemia. Clinical variables do not have adequate sensitivity and specificity to accurately predict which children with AGE have hypoglycemia. Glucose determination should be considered for these patients. PMID- 14552525 TI - Urinary tract infection due to Salmonella species in children/adolescents. PMID- 14552526 TI - Neonatal myocarditis presenting as an apparent life threatening event. PMID- 14552527 TI - Risk factors for tobacco use by utilizers of the pediatric emergency department. PMID- 14552528 TI - A "hickey". Epidural brain abscess, osteomyelitis of the frontal bone, and subcutaneous abscess (pott puffy tumor). PMID- 14552529 TI - The optimizing action of the synthetic peptide Selank on a conditioned active avoidance reflex in rats. AB - The actions of the synthetic heptapeptide preparation Selank on learning and memory processes in rats with initially low levels of learning ability were compared with those in normal rats, using a method based on acquisition of a conditioned active avoidance reflex, with repeated administration of peptide 15 min before the start of training sessions for four days. The effects of Selank (300 microg/kg) were compared with the effects of the nootrope piracetam (400 mg/kg). These experiments showed that Selank significantly activated the learning process in rats with initially poor learning ability, with effects apparent after first dose on training day 1. The effect progressively increased on repeated administration of Selank: the total number of correct solutions increased and the number of errors decreased (p < 0.05). The maximum optimizing activity of Selank on learning in normal rats was seen on day 3 of repeated administration and training, i.e., after formation of the initial consolidation phase. The dynamic features of the development of the activating action of Selank and piracetam were described. Comparison of the results obtained here with data on the anti-anxiety actions of Selank suggested potential for its use in optimizing mnestic functions in conditions of elevated emotional tension. PMID- 14552531 TI - Dynamics of neuron activity in the lateral preoptic area of the hypothalamus during the sleep-waking cycle. AB - Chronic experiments were performed on cats to study neuron spike activity in the lateral preoptic region of the hypothalamus in active and calm arousal and in the slow-wave and paradoxical phases of sleep. The dynamics of spike frequencies and the patterns of activity in the sleep-waking cycle allowed neurons to be divided into three populations. Cells showing increases in the frequency of single spikes as the level of consciousness decreased, on the transition to slow-wave sleep and then to the paradoxical phase of sleep were assigned to the "anti-waking" system, which, being a component of the somnogenic system of the brain, is involved in the mechanisms initiating and increasing the depth of sleep by inactivating the arousal-maintaining system. Cells with maximum spike frequencies in light, slow wave sleep and demonstrating single and train discharges in association with "sleep" spindles, were regarded as elements of the system responsible for forming this state. The remaining neurons had activity characteristics which were similar in the active arousal state and paradoxical sleep and decreased their spike frequencies in calm arousal and the slow-wave phase of sleep in parallel with the transition from the continuous-arithmetic to the mixed type of activity. Changes in the activity of this type of cell during the sleep-waking cycle appear to reflect rearrangements in controlling influences from the somnogenic and arousal systems of the brain. PMID- 14552530 TI - Monitoring of the oxidation-reduction state of brain structures in freely moving rats during sleep-waking cycles by potentiometric recording. AB - Freely mobile mongrel male rats weighing 300-350 g were used for studies of changes in the oxidative-reductive (redox) state of brain tissue during cycles of waking, slow-wave sleep, and paradoxical sleep, by recording the potential of the oxidative-reductive state of brain tissue with platinum electrodes implanted into the cerebral cortex ad hippocampus. Electromyograms were also recorded from the cervical muscles, and overall movement activity was also recorded. A common platinum reference electrode was implanted into the nasal bones. These experiments showed that in rats, episodes of waking and paradoxical sleep occurred on the background of increases in the oxidation-reduction potential state of brain tissue at a series of brain points, which we termed "metabolically active." Transitions from waking and paradoxical sleep to slow-wave sleep were accompanied by decreases in the potential of the redox state. The magnitude of changes in the tissue redox state varied up to 100 mV. It is suggested that transitions from waking and paradoxical sleep to slow-wave sleep are accompanied by dynamic changes in the balance of brain tissue energy metabolism between the main energy sources. Oxidative phosphorylation dominates in waking and paradoxical sleep, while aerobic glycolysis dominates slow-wave sleep. We suggest that this latter should be interpreted as a decrease in the potential of the tissue redox state and the formation within the tissue of oscillations during slow-wave sleep. Formation of oscillations is typical for acceleration of glycolytic processes. Recently published data suggest that the major compartment or aerobic glycolysis is the astroglia. PMID- 14552532 TI - Changes in the signal value of a conditioned stimulus during acquisition of a conditioned two-way avoidance reflex in rats. PMID- 14552533 TI - The polysynaptic spine-stem reflex in clinical neurology. AB - Data from 25 years of studies of the spinobulbospinal (SBS) reflex in normal conditions and in a variety of nervous system diseases with lesions of the afferent and efferent systems, subcortical brain formations, and musculature (contracture of the expressive muscles, myofascial pain syndrome) are presented. The instability of many characteristics of the reflex (latency, amplitude, duration of response, presence) is shown to characterize this reflex in normal conditions. These measures become more stable in central nervous system pathology, which is evidence for impairment of programs which construct and execute movements. Lesions of the peripheral neuromotor apparatus produced no significant changes in the characteristics of the SBS reflex. PMID- 14552534 TI - Mismatch negativity in evoked brain potentials in adolescents in normal conditions and attention deficit in response to presentation of short-duration acoustic stimuli. AB - Experiments were performed to measure the critical duration of acoustic stimuli at which the generation of mismatch negativity is still possible in auditory evoked potentials in the human brain, arising in conditions of presentation of standard and deviant stimuli differing in terms of frequency (1000 and 1200 Hz respectively). Studies were performed in two groups of adolescents (aged 11-14 years): 1) those with normal attention characteristics and 2) children with attention deficit. The results showed that normal children had the same critical duration for sound stimuli at which mismatch negativity could be generated as adults. Mismatch negativity still appeared at a stimulus duration of 50 msec, while the mechanism generating mismatch negativity did not operate at durations of 30 and 11 msec. In children with attention deficit, statistically significant mismatch negativity was not seen at all durations of short acoustic stimuli (11, 30, and 50 msec). PMID- 14552535 TI - Neural correlates of the results-of-action acceptor in a functional biotechnical complex. AB - This report presents data illustrating the neurophysiological features of efferent-afferent convergence on cortical neurons. During combination of stimulus of pyramidal tract axons with electrocutaneous reinforcement, some of the study neurons were found to change the parameters of their activity evoked by stimulation of this tract: evoked responses started to reproduce the structure of responses to the reinforcement. The most dynamic rearrangements of pyramidal tract responses were obtained in experiments in which the study neurons were included in a biotechnical complex with feedback, the complex consisting of "neuron-computer-stimulator-animal" and actually being an analog of a natural functional system. The role of efferent-afferent convergence on CNS neurons in the development of the results-of-action acceptor of a functional system for a voluntary behavioral act is discussed. PMID- 14552536 TI - Comparison of the postural and movement components during learning by dogs of an operant defensive reflex. AB - Chronic experiments on six dogs using a model of an operant defensive reflex associated with maintenance of flexion of the hindlimb of specified amplitude were performed to compare the characteristics of the postural and movement components during the learning process. Dogs were placed standing on four tension platforms. Signals were activated and data were recorded and stored using original PC programs. Original programs running on another PC were used to analyze the data. All dogs showed a series of characteristics for the appearance of a diagonal pattern of conditioned reflex posture rearrangement. During the period of complete formation of the "coordinated" program of the operant reflex (indicated by high performance criteria for execution of the operant task), the diagonal pattern of posture rearrangement was seen extremely rarely, in only occasional performances in calm dogs. It was only during the period of complete automatization of the movement habit that the diagonal pattern of postural rearrangement was seen consistently in all performances. By this time, there was a sharp increase in the tensogram amplitudes for all four limbs, which sharp increases in the correlation coefficients between individual tensogram performances. These results suggest that in the experimental conditions used here, the diagonal pattern of postural rearrangement appeared significantly later than the movement pattern needed for resolving the operant task. These data also lead to the conclusion that dogs can complete operant defensive limb movements associated with maintaining a specified flexor posture in the absence of preliminary rearrangement of the posture having the diagonal pattern. PMID- 14552537 TI - Rhythmic photostimulation and the number of alpha-rhythm dipoles in the human brain. AB - The numbers of alpha-rhythm equivalent current dipoles (ECD) arising in the human brain before and during rhythmic photostimulation at the alpha-rhythm frequency was studied in six healthy adult subjects. Dipoles were calculated using a single dipole model for the whole of the alpha-range and three subranges by solution of inverse equations in a three-layer model of the head obtained by simultaneous use of EEG data and MRI tomograms of the subjects' heads. The number of apparent ECD was significantly associated with rhythmic photostimulation and depended on the phase of the alpha-rhythm wave at which stimulation started and on the type of visual illusion (circle, spiral, grid) appearing during this time. The relationship between these data and the hypothetical wave process scanning the human visual cortex at the frequency of the alpha-rhythm is discussed. PMID- 14552538 TI - Spatial distribution of coefficients of asymmetry of brain bioelectrical activity during the experiencing of negative emotions. AB - Significant differences in the spatial distribution of the coefficients of asymmetry of the power of bioelectrical activity were identified in negative emotions of different qualities (anger and grief). In the case of the sthenic (excitable) emotion anger, there was an increase over baseline in the positive value of the coefficient of asymmetry in the anterior areas and the beta2 range, while for the asthenic emotion grief, there was an increase in the negative value of the coefficient in the beta1 range and a generalized increase in slow-wave activity. PMID- 14552539 TI - Structure of reciprocal connections of visual cortical fields 17 and 18 in the cat. AB - Microiontophoretic application of horseradish peroxidase to individual columns in fields 17 and 18 of the cat cortex was used to identify the distribution by area and layer of retrograde labeled cells in both fields. After application of marker to fields 17 or 18, the area of labeled cells in field 17 (in the tangential plane) was extended and orientated along the projection of the horizontal meridian of the field of vision. The area of labeled cells in field 18 in these cases was orientated along the projections of the vertical meridian. Similar differences in the organization of the connections of fields 17 and 18 were seen in the projection zone of the central 10 degrees of the field of vision at different elevations. Thus, the spatial distributions of internal and external connections in each field coincide and their orientations in fields 17 and 18 are mutually perpendicular. It is suggested that field 17 performs the more detailed analysis of information on the horizontal components of an image and communicates this to field 18, while field 18 is responsible for the more detailed analysis of information about the vertical components of the same image, communicating this to field 17. PMID- 14552540 TI - Corticoliberin protects neurons from the negative influences of "dysfunctins" in living olfactory cortex slices. AB - The protective effects of corticoliberin on living rat olfactory cortex slices during perfusion with "dysfunctins" extracted from cerebrospinal fluid of drug addicts were studied. Isolated perfusion of slices with medium containing "dysfunctins" led to irreversible suppression of the amplitude of individual components of focal potentials induced by electrical stimulation of the lateral olfactory tract. The maximum level of depression was seen for the AMPA and NMDA components of EPSP. Preliminary perfusion of slices with medium containing corticoliberin (100 nM) for 15 min partially, and for 30 min completely protected processes mediated by activation of AMPA and NMDA receptor mechanisms from the negative influences of "dysfunctins." It is suggested that corticoliberin can induce its protective effects either via its own specific receptors or non specifically via glutamate receptors. It is also possible that both of these mechanisms act in combination. PMID- 14552541 TI - Effects of weightlessness and movement restriction on the structure and metabolism of the soleus muscle in monkeys after space flight. AB - After humans and animals have been in conditions of real and modeled weightlessness, the most marked changes are seen in the "slow" tonic muscles, particularly soleus. Studies of the effects of weightlessness and movement restriction on the soleus muscle in monkeys demonstrated significant reductions in the sizes of slow and rapid fibers due mainly to the actions of real weightlessness (rather than movement restriction in the space capsule). Protein loss in soleus muscle fibers in monkeys following space flight was more marked than loss of other components, including water. The level of atrophy of soleus muscle fibers in these conditions was greater than the decrease in the number of capillaries. Succinate dehydrogenase activity in soleus muscle fibers decreased proportionally to the reduction in fiber size. PMID- 14552542 TI - Potential of phosphorus nuclear magnetic resonance spectroscopy in studies of the energy metabolism of skeletal muscles. AB - The aim of the present study was to investigate the possibility of phosphorus magnetic resonance spectroscopy (MR spectroscopy) in the diagnosis of metabolic lesions of skeletal musculature in patients with intermittent claudication syndrome, chronic cardiac failure, and varicose diseases of the lower limbs. Studies included 50 males: 20 patients with intermittent claudication, 10 patients with chronic cardiac failure, and 10 patients with varicose veins. The control group consisted of 10 healthy volunteers. The following measures were determined: the phosphocreatinine index, the intracellular pH in the gastrocnemius muscle, and the half-recovery time for the phosphocreatinine index. The phosphocreatinine index and the pH at rest did not differ between study groups. Isotonic exercise produced no change in the phosphocreatinine index in the control group; patients with intermittent claudication showed a 26.1% decrease, patients with chronic cardiac failure showed an 8% decrease, and patients with varicose veins showed a 25.6% decrease. The only group showing a significant decrease in pH during exercise was the group of patients with intermittent claudication. This group also showed an inverse correlation between the pressure index and the extent of the decrease in the phosphocreatinine index. Thus, MR spectroscopy provides a non-invasive diagnostic method for lesions of energy metabolism in skeletal musculature in patients with deranged peripheral hemodynamics. PMID- 14552543 TI - The effects of serotonin on the differentiation of neurons producing vasoactive intestinal polypeptide in the suprachiasmatic nucleus of the rat. AB - The morphogenetic influences of serotonin on the differentiation of neurons synthesizing vasoactive intestinal polypeptide (VIP) in the suprachiasmatic nucleus were studied in rats. This was addressed by comparative morphofunctional analysis of VIP neurons in adult rats whose brains developed prenatally in conditions of normal and deficient serotonin metabolism. Serotonin deficiency was created in fetuses by treatment of their mothers with p-chlorophenylalanine (PCPA). Pregnant females in controls were treated with 0.9% NaCl. VIP neurons in experimental and control animals were found to show no differences in VIP mRNA concentrations and, probably, in the level of VIP synthesis. However, inhibition of serotonin synthesis led to an increase in the number of VIP-immunoreactive neurons and an increase in the VIP concentration within these cells. This was not associated with any change in neuron size, which was an indicator of the absence of functional hypertrophy accompanying activation of specific synthesis. Comparison of the data obtained here showed that during prenatal ontogenesis, serotonin has an imprinting influence on the differentiation of VIP neurons and is probably involved in the formation of the mechanism of VIP secretion. PMID- 14552544 TI - Characteristic of changes in the structure and metabolism of the vastus lateralis muscles in monkeys after space flight. AB - Monkeys subjected to space flight were found to have significant decreases in the sizes of slow and rapid fibers in the vastus lateralis muscle, due not only to weightlessness but also, to some extent, to restriction of movement activity within the capsule. The quantity of total protein in muscle fibers did not decrease. The respiratory peak in the pool of vastus lateralis muscle fibers decreased after space flight, as did the activity of oxidative enzymes (particularly in rapid fibers of the vastus lateralis muscle). PMID- 14552545 TI - A device for the stereotaxic fixation of the spine in animals. PMID- 14552547 TI - Studies of selective attention in dogs using the coherence-phase characteristics of cortical potentials over a wide range of frequencies, 1-220 Hz. AB - The state of selective attention formed during operant food-related behavior was studied using the coherence-phase characteristics between potentials in several areas of the neocortex at frequencies of 1-220 Hz. Functional groups were identified among the areas compared, which appear to have priority for this state. The temporal relationships between potentials in these groups were established from values for coherence functions at a particular optimum level (0.7) mainly in the band 1-15 Hz and the high-frequency range 40-200 Hz. The phenomenon of synchronicity appeared at phase shifts close to zero, while at significant phase shifts the phenomenon of non-synchronous time relationships with defined spatial directions was seen in the high-frequency range. PMID- 14552546 TI - A possible mechanism for the effect of neuromodulators and modifiable inhibition on long-term potentiation and depression of the excitatory inputs to hippocampal principal cells. AB - A postsynaptic mechanism for the influences of various neuromodulators and modifiable disynaptic inhibition on long-term potentiation and depression of the excitatory inputs to granule and pyramidal neurons in the hippocampus is described. According to this mechanism, facilitation of the induction of long term depression/potentiation at the excitatory input to the inhibitory interneuron induced by the action of a neuromodulator on a receptor bound to a G(i/0)/(Gs or G(q/11)) protein can lead to decreases/increases in GABA release, weakening/strengthening of the inhibitory action on the target cell, and improvement in the conditions for induction of long-term potentiation/depression of the excitatory input to this cell. In the absence of inhibition, the same neuromodulator, activating the same type of receptors on the target cell, would facilitate induction of long-term depression/potentiation in that cell. The resultant effect of the action of the neuromodulator on the target cell depends on the ratio of the "strengths" of the excitatory and inhibitory inputs to the cell, on the presence on the interneuron and the target cell of the same or different types of receptors sensitive to this neurumodulator, and on the concentration of the neurumodulator, because of its different affinities for the receptors through which its differently directed effects on postsynaptic processes are mediated. Predictions based on this mechanism are in agreement with known experimental data. PMID- 14552548 TI - Organization of neural networks in the neocortex. AB - Implanted semimicroelectrodes were used in conscious cats to record spike discharges from groups of close-lying neurons, i.e., multineuron activity, in the deep layers of the frontal and motor areas of the cortex at different levels of food motivation. Spike activity was extracted from 4-7 neurons and interneuronal interactions were studied by cross-correlation analysis between neighboring neurons in each zone (local networks) and between neurons in two zones (distributed networks) with analysis epochs of 0-100 msec. The results showed that neurons in local networks can be divided into two subgroups: neurons with high-amplitude spikes and a predominance of output (divergent) connections and neurons with low-amplitude spikes and a predominance of input (convergent) connections. Local networks are based on powerful monosynaptic connections (with delays of up to 2 msec) between large and small neurons. Most connections in distributed networks were between small neurons in local networks of the frontal cortex and large neurons in local networks in the motor cortex. Food deprivation for 24 h mainly affected late (with delays of 2-100 msec) cross-correlation interneuronal relationships in both local and distributed networks. PMID- 14552549 TI - A computer model of neural processes observed in the cat motor cortex during performance of an operant movement. AB - This report describes a computer model of a "column" in the cat motor cortex. The model includes two layers of two-segment pyramidal neurons with two groups of inhibitory interneurons in each layer, which selectively control the somatic and dendritic segments of the pyramidal cells. In this model, neurons include active sodium, calcium, and several types of potassium currents. Excitatory connections between neurons are of the AMPA and NMDA types, while collateral connections between neurons of the upper layer are mainly of the NMDA type; connections between neurons in the lower layer are of the AMPA type. All inhibitory connections are of the GABA(A) type. The model reproduces the main neuronal processes seen in the cat motor cortex during performance of an operant movement. Pyramidal neurons of the upper layer generate primary and secondary responses to external stimuli. As in real experiments, secondary NMDA-dependent responses appear when GABA(A) inhibition is weakened and disappear when stimulation is increased; these properties of secondary responses are only reproduced when NMDA receptors are located in the terminals of collateral connections. Using only rapid NMDA-independent connections, neurons in the lower layer generate a slow bell-shaped wave of excitation (a "motor command"), which is formed by sequential activation of neurons with dendritic trees of different sizes. PMID- 14552550 TI - Cholinergic modulation of neuron spike responses to dendritic and somatic application of excitatory amino acids. AB - The effects of acetylcholine on the spike discharges of neurons induced by iontophoretic application of excitatory amino acids to the bodies and dendrites of cells were studied in 98 neurons in living slices of guinea pig parietal cortex. Acetylcholine applied microiontophoretically to both the bodies and dendrites facilitated improvements in the parameters of responses induced by dendritic activation, with significant decreases in latent periods and increases in the intensity and duration of responses. Thee effects were stably induced at distances of 300 microm from the body and lasted 1 min after exposure to acetylcholine ended. Responses induced by application of excitatory amino acids directly to the cell body did not change significantly in the presence of acetylcholine regardless of the point on the membrane at which they were applied. It is concluded that the predominant effect of acetylcholine is on the efficiency of dendrosomatic conduction. PMID- 14552551 TI - Comparative analysis of the frequency of neuron spike activity in the sensorimotor cortex of the right and left hemispheres in conditions of immobilization catatonia in rabbits. AB - Multineuron activity was recorded from the sensorimotor cortex of the right and left hemispheres during immobilization catatonia in rabbits. The first session of immobilization of the animals was followed by changes in spike frequency in 47% of neurons in the sensorimotor cortex of the right hemisphere. Of these, 30% showed decreases in spike frequency and 17% showed increases. Spike frequency in the sensorimotor cortex of the left hemisphere changed in only 18% of cells, of which 13% showed decreases in spike frequency and 5% showed increases. The spike frequency of neighboring (recorded with the same electrode) neurons could change reciprocally. Differences in neuron activity in the two hemispheres were virtually absent after the second session of immobilization (several days after the first)--spike activity changed in 21% of neurons in the right hemisphere and 24% in the left hemisphere. The ratios of the numbers of neurons with increases and decreases in spike activity in "hypnosis" also became identical in the cortex of the right and left hemispheres. A hypothesis is proposed for the involvement of cortical neurons in the organization of "hypnosis-like" states. PMID- 14552552 TI - The effects of anxiogenic stress on glucocorticoid sensitivity, glucose tolerance, and alloxan resistance in rats. AB - Four sessions of immobilization stress induced anxiogenic behavioral disorders in rats, which were accompanied by decreases in glucocorticoid sensitivity, increases in MAO-B activity in brain tissue, increases in tolerance to glucose loading, and decreases in resistance to acute hypoxia. Alloxan diabetes was accompanied by a decrease in behavioral activity of rats in the open field test, with an increase in cerebral MAO-B activity. Preceding anxiogenic stress increased the extent of the alloxan-induced increase in cerebral MAO-B activity and the accompanying abnormalities in the rats' behavior, and also potentiated the hyperglycemic effect of alloxan. PMID- 14552553 TI - Modification of redox sites of N-methyl-D-aspartate receptors affects anoxia induced changes in the bioelectrical activity of rat brain olfactory cortex slices. AB - Living slices of Wistar-Kyoto rat brain olfactory cortex were used to study the effects of the thiol-oxidizing agent 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), which inhibits NMDA receptor activity, on changes in the generation of evoked focal potentials (NMDA and non-NMDA EPSP) in response to long-term and short-term anoxia, which induces functional damage and facilitates increases in the resistance of neurons to severe hypoxia respectively. These studies showed that DTNB (200 microM) efficiently prevented the suppression of focal EPSP generation due to long-term anoxia in most slices. In addition, DTNB partially reversed the protective effect of preconditioning with short-term anoxia on the impairment of focal EPSP generation induced by long-term anoxia. This affected the NMDA component of the EPSP to a greater extent than the non-NMDA component. The possible role of changes in the state of modulatory redox sites of NMDA receptors in the mechanisms of functional damage and increases in neuron resistance due to hypoxia is discussed. PMID- 14552554 TI - The effects of ante- and postnatal hypoxia on the central nervous system and their correction with peptide hormones. AB - Ante- and postnatal hypoxia significantly worsened the postnatal development of animals. The posthypoxic behavioral model included hyperactivity and decreased learning ability, these being typical manifestations of attention deficit disorder. A peptide constellation prevented and significantly improved posthypoxic postnatal development and eliminated the majority of negative behavioral changes. PMID- 14552556 TI - Electrophysiological studies of the effects of 5,6-dihydroxytryptamine on the acquisition of a conditioned defensive reflex in snails. AB - The role of serotonin in the membrane properties of identified neurons was studied during acquisition of a conditioned defensive reflex. Serotonin deficiency was created using the neurotoxic serotonin analog 5,6 dihydroxytryptamine (5,6-DHT). Injection of 5,6-DHT was found to prevent decreases in the membrane and threshold potentials of command neurons during acquisition of the conditioned reflex as compared with snails given 5,6-DHT without training. Common snails recovered the ability to acquire the conditioned reflex two weeks after a second 5,6-DHT injection. PMID- 14552555 TI - Transmitter secretion in the frog neuromuscular synapse after prolonged exposure to calcium-free solutions. AB - Experiments on neuromuscular synapses from frog skin/chest muscle preparations in conditions of extracellular recording addressed changes in the spontaneous and evoked transmitter secretion after long-term (1.5-6 h) maintenance of preparations in calcium-free solution containing EGTA. Use of three microelectrodes for recording of single-quantum postsynaptic signals showed that calcium-free solution altered the characteristic topography of transmitter secretion in nerve terminals, with widening and fusion of groups of transmitter release. These changes persisted after preparations were returned to the initial solution. These data suggest that calcium-free solutions lead to disorganization of the active zones of nerve endings, At initially low extracellular Ca ion concentrations (0.15-0.4 mM), disorganization of active zones induced by prolonged maintenance of preparations in calcium-free solutions led to decreases in the mean amplitude of endplate currents (EPC) because of decreases in their quantum composition, increases in the time course of transmitter secretion, and decreases in the frequency of miniature endplate currents. The relationship between quantum composition of EPC and the extracellular Ca ion concentration showed a sharp displacement towards higher concentrations, without significant changes in the slope of the relationship. At high initial Ca concentrations (1.8 mM), long-term exposure to calcium-free solutions led to a less marked decrease in EPC amplitude. It is suggested that the extra- and intracellular Ca ion concentrations support the maintenance of the characteristic morphofunctional organization of the apparatus responsible for transmitter secretion in frog nerve endings. Disorganization of the active zones leads to disruption of elements involved in transmitter secretion and decreases in the efficiency of secretion. PMID- 14552557 TI - Involvement of dopaminergic processes in the striatum during the effects of corticoliberin on the behavior of active and passive rats. AB - The effects of intranasal corticoliberin on behavior in the open field test were studied in rats with active and passive behavioral strategies (lines KHA and KLA); levels of dopamine and noradrenaline and their metabolites were measured in the striatum and hypothalamus. In KLA rats, administration of the neurohormone led to increases in motor and investigative activity, while decreases were seen in KHA rats. There were no interline differences in catecholamine levels in the hypothalamus, while dopamine levels in the KLA striatum nearly doubled and metabolite levels (DOPAC, HVA) were significantly lower than in KHA rats. Corticoliberin increased dopamine and noradrenaline levels in the hypothalamus of both rat lines, with significant decreases in the striatum. This decrease was more marked in KLA rats, probably due to the faster metabolism of transmitters in the presence of neurohormones, as indicated by the increase in metabolite levels in this structure. PMID- 14552559 TI - Using routinely collected data from a National TB Programme. PMID- 14552558 TI - Changes in hippocampal excitability after exclusion of amygdalohypothalamic connections. PMID- 14552560 TI - The treatment of tuberculosis in South Korea. AB - South Korea's complex system of tuberculosis control has never been fully described. The prevalence of tuberculosis has dropped dramatically since 1965, partly because of farsighted governmental policy that provided low-cost, accessible tuberculosis treatment to the entire population. Within the tuberculosis control system, public and private sector entities provide a wide variety of treatment options. The National Tuberculosis Program focuses on improving cure rates for new cases, while the private sector has taken more of a role in the treatment of drug-resistant tuberculosis and other types of complicated cases. There has been a decrease in drug-resistant tuberculosis since 1980 for multiple reasons, including increased cure rates from the introduction of rifampin-based regimens, improved nutrition and living standards, and the treatment of drug-resistant cases in the private sector. Multidrug-resistant tuberculosis, however, still poses a significant threat to public health. The limited outcomes data that exist in South Korea for multidrug-resistant tuberculosis treatment suggest that cure rates are low and failure and abandonment rates are high. New public health measures are needed to improve the control of multidrug-resistant tuberculosis. PMID- 14552561 TI - Tuberculosis control in Samara Oblast, Russia: institutional and regulatory environment. AB - SETTING: Tuberculosis control programme in Samara Oblast, Russia, funded in part by the government of the United Kingdom. OBJECTIVE: To identify and evaluate institutional and regulatory influences as well as incentives and disincentives that might be amenable to change in the promotion of the DOTS strategy. DESIGN: Multidisciplinary situational analysis through in-depth interviews of stakeholders and review of official federal and oblast documents. RESULTS: Interpretation of traditional notification data is complex because classification and reporting systems differ from World Health Organization principles. Regulations governing financing encourage lengthy hospitalisations and interventions, and provide few incentives to shift policy to ambulatory care. CONCLUSION: Accurate comparability of epidemiological trends and programmatic successes requires equivalent classification and reporting systems. If the DOTS strategy is to be sustainable, changes to financing systems will be needed. PMID- 14552562 TI - Tribal differences in perception of tuberculosis: a possible role in tuberculosis control in Arusha, Tanzania. AB - SETTING: Arusha, Tanzania. OBJECTIVE: To determine tribal differences in knowledge and practices that might influence tuberculosis control. METHOD: Twenty seven villages were selected randomly out of 242 villages in four districts. In each village, a general and a livestock keeping group were selected at random. The households were home-visited and 426 family members were interviewed. RESULTS: On average, 40% of respondents practised habits that might expose them to both bovine and human tuberculosis. The Barabaig tribe had a significantly higher number of respondents (50%, chi2(2) = 5.1, P = 0.024) who did not boil milk. Eating uncooked meat or meat products was practised by 17.9% of all respondents. The habit was practised more by Iraqw (21.1%, chi2(2) = 6.9, P = 0.008) and Barabaig (31.6%, chi2(2) = 5.6, P = 0.016) than other tribes. About 75% of the respondents had a poor knowledge of tuberculosis. CONCLUSION: All tribes had habits and beliefs that might expose them to both bovine and human tuberculosis. The Iraqw and Barabaig tribes practised such habits more than other tribes. Knowledge of tuberculosis was limited in all tribes. PMID- 14552563 TI - Clinical and radiological presentation of 340 adults with smear-positive tuberculosis in The Gambia. AB - SETTING: Four clinics in The Gambia. OBJECTIVE: To document clinical and radiographic presentations of sputum smear-positive tuberculosis in adults. DESIGN: Newly diagnosed acid-fast bacilli (AFB) smear, culture-positive tuberculosis patients aged > or = 15 years were interviewed and examined, and underwent tuberculin skin testing, HIV testing and chest X-ray reviewed by a chest physician using set criteria. RESULTS: Of 340 patients enrolled (median age 29 years; males 73%), 8.3% were HIV-positive. One-third reported haemoptysis, > 90% reported weight loss and fever, and wasting was the most common sign (69%). Crepitations were the most frequent auscultatory finding (41%). The most common radiological lesion was a patchy infiltrate (> 90%). Cavitation was present in 206 patients (60.6%), most frequently occurred in the upper lung fields, was associated with increasing bacterial load in the sputum, and was less prevalent in HIV-positive patients (45% vs. 62%; P = 0.07). Auscultatory and chest X-ray findings matched only one-third of the time. CONCLUSION: In our setting, wasting is the most common clinical sign of sputum smear-positive tuberculosis. Auscultatory findings correlate poorly with radiological abnormalities. Cavitation is associated with increasing bacterial load in the sputum, and is therefore a strong indicator for early treatment. PMID- 14552564 TI - Characteristics, management and outcome of patients with recurrent tuberculosis under routine programme conditions in Malawi. AB - SETTING: All 43 non-private hospitals in Malawi, which registered TB cases between 1 July 1999 and 30 June 2000. OBJECTIVES: To determine 1) the characteristics, management and treatment outcome, 2) timing of the previous episode of TB, and 3) pattern of drug resistance in patients registered with recurrent smear-positive pulmonary TB. DESIGN: Retrospective data collection using TB registers and laboratory culture and drug sensitivity registers. RESULTS: There were 748 recurrent patients; data were available for 747. Of these, 487 (65%) successfully completed a re-treatment regimen, 185 (25%) died and the remainder had another outcome. Information about previous TB was recorded for 491 (66%) patients. In 286 (58%) there were 2 years or less between completing and re-starting treatment. Only 307 (41%) patients had sputum sent for culture and drug sensitivity tests. In 164 patients with cultures of Mycobacterium tuberculosis, 122 (81%) were fully sensitive, 25 (15%) had resistance to isoniazid and/or streptomycin, and 6 (4%) had resistance to isoniazid and rifampicin (MDR-TB). CONCLUSION: Patients with recurrent TB had acceptable treatment outcomes, and most had fully sensitive organisms. Over half had recurrent TB 2 years or less after completing treatment. Ways to prevent recurrence need to be investigated and implemented in the field. PMID- 14552565 TI - Defining and assessing the maximum number of visits patients should make to a health facility to obtain a diagnosis of pulmonary tuberculosis. AB - SETTING: All non-private hospitals in Malawi. OBJECTIVES: To determine 1) how many patients with pulmonary tuberculosis (PTB) exceed the maximum number of visits needed for registration as defined by the National Tuberculosis Control Programme, and 2) the factors associated with this delay. DESIGN: Cross-sectional study interviewing hospitalised patients with new smear-positive and smear negative PTB. RESULTS: Of 380 patients with PTB admitted to the 44 hospitals visited between April and June 2002, 329 (212 smear-positive and 117 smear negative PTB) were interviewed: 64 (30%) smear-positive PTB patients needed more than five visits, and 44 (37%) smear-negative PTB patients needed more than six visits before being registered and started on treatment. Factors associated with exceeding the maximum number of visits were the first visit being to a health centre, submission of > 1 set of sputum specimens, and > 1 course of antibiotics. The main consequence of exceeding the maximum number of visits was increased duration of cough and increased time spent at health facilities. CONCLUSION: One third of patients exceed the maximum number of visits for registration of PTB. The main consequence of this is an increased duration of cough and an increased time spent at health facilities. Ways to reduce this delay need to be found. PMID- 14552566 TI - Screening and management of tuberculosis in immigrants: the challenge beyond professional competence. AB - SETTING: Right of entry may sometimes be denied to immigrants because of the threat of tuberculosis. During 1990-2000 some 1050000 immigrants, mostly from countries highly endemic for TB, arrived in Israel, a low prevalence country. Nevertheless, TB rates in Israel have remained low. OBJECTIVE: To emphasise the challenge beyond technical competence for TB control for immigrants from the perspective of Israel's National Tuberculosis Programme (NTP). MATERIALS AND METHODS: We defined criteria for an NTP geared to immigration, and analysed our implementation of the European Task Force recommendations on international migration and TB control. We interviewed immigrants and health care workers to identify barriers to diagnosis, prevention and treatment of TB among immigrants. We used classical epidemiology to evaluate the impact of immigration on TB rates in the host population. RESULTS: Until now there has been no evidence of significant spread of TB from immigrants to the host population. Successful outcome of treatment has been noted in over 75%, although a sub-population of immigrant substance abusers is proving more difficult to treat. CONCLUSIONS: The risk of TB for the host country is very low and it seems possible to enhance TB control in immigrants with measures designed to address their cultural needs. PMID- 14552567 TI - Clustering of tuberculosis among senegalese immigrants in Italy. AB - OBJECTIVE: To study clustered Mycobacterium tuberculosis isolates as an indicator of recent TB transmission in a small urban setting in Italy, and to determine associated risk factors. METHODS: M. tuberculosis strains isolated between 1991 and 1997 were characterised by IS6110 restriction fragment length polymorphism (RFLP) analysis. RESULTS: One hundred and ninety-five isolates were available for RFLP analysis, which revealed 163 different patterns. Available cases were represented by 137 Italians (70%), 32 Senegalese (17%), and 26 other foreign-born cases (13%). A unique fingerprint pattern was found in 143 cases (73.3%), while 52 strains (26.7%) were grouped into 20 clusters. Nineteen cases (10%) were resident in the same quarter of Brescia with a high density of Senegalese immigrants (Area A). An increased probability of yielding clustered M. tuberculosis strains was associated with residence in Area A (OR 3.87, 95%CI 1.42 10.56; P = 0.02) and being Senegalese (OR = 5.96, 95%CI 1.48-23.97; P = 0.005). In the logistic regression analysis, being Senegalese was independently associated with yielding a clustered M. tuberculosis strain. CONCLUSIONS: Our results demonstrate a clustering of TB cases among Senegalese immigrants and suggest that RFLP analysis may be used to identify geographical areas where efforts can be targeted to interrupt TB transmission. PMID- 14552568 TI - Isoniazid resistance among tuberculosis patients by birth cohort in Germany. AB - SETTING: Germany, 1997-2000. OBJECTIVE: To determine the prevalence of isoniazid resistance by year of birth among previously treated and untreated tuberculosis patients by country of birth. DESIGN: Univariate and multivariate analyses of data on 8658 cases from a nationally representative sample. RESULTS: The crude prevalence of isoniazid resistance was 2.4% and 9.9% among Germany-born patients respectively without and with a prior treatment history. Among those without a history, there was virtually no difference between quartiles of birth cohorts, while among those with, a large decrease from the oldest to the youngest was observed. Among foreign-born patients, the prevalence of isoniazid resistance increased from 3.8% to 11.8% from the oldest to the youngest among those without, and more than doubled from less than 20% to more than 40% among those with prior treatment. CONCLUSION: This analysis suggests a remarkably constant and low prevalence of isoniazid resistance among never treated Germany-born patients. In contrast, among the diverse group of foreign-born patients, the prevalence increases appreciably from the oldest to the youngest birth cohorts. This indicates that tuberculosis treatment policies in Germany have been sound and that great care is needed to prevent acquisition of additional resistance among foreign-born patients. PMID- 14552570 TI - Extrathoracic tuberculosis lymphadenitis in adult HIV seronegative patients: a population-based analysis in Houston, Texas, USA. AB - OBJECTIVE: To evaluate the covariates associated with extrathoracic tuberculosis lymphadenitis (ETBL) among adult HIV-seronegative patients. METHODS: Enrollees were interviewed for TB risk assessment, their medical records were reviewed, and their Mycobacterium tuberculosis isolates underwent molecular characterization. Between 1 October 1995 and 30 September 1999, HIV-negative patients with ETBL were compared with other HIV-negative TB patients. RESULTS: We identified 73 ETBL cases (5%) out of a total of 1371 adult HIV-negative enrollees. Significant variables predicting ETBL in the univariate analysis included age < 45 years, female sex, Asian ethnicity, foreign birth, BCG vaccination, and infection with a M. tuberculosis isolate identified in major genetic group 1. Further analysis by birth country revealed increased ETBL risk for persons from countries other than the Americas and with a TB incidence > 25 per 100 000 per year. The multivariate model demonstrated increased risk for ETBL for patients of female sex (OR = 2.6, P < 0.01) and birth in Africa or South-east Asia (OR = 4.8; P = 0.03 and OR = 33.6; P = 0.01, respectively). CONCLUSIONS: In adult HIV-negative patients, ETBL occurs more frequently in females and in immigrants from countries other than the Americas; persons from India, South-east Asia and the Eastern Mediterranean exhibited the highest risk among these regions. PMID- 14552569 TI - Smoking and tuberculosis in Hong Kong. AB - OBJECTIVE: To study the relationship between smoking and tuberculosis in Hong Kong. METHOD: Indirect sex and age adjustment was used to compare the prevalence of ever smokers between a sample of 851 patients from the 1996 tuberculosis notification registry and the general population. The clinical characteristics of smokers and non-smokers were compared by stratified univariate analysis and multiple logistic regression. RESULTS: Tuberculosis patients were more likely to have smoked than population controls. The respective odds ratios for ever smoking between tuberculosis patients and population controls were 2.44 and 2.08 for males and females aged 16-64 (Mantel-Haenszel weighted OR = 2.40, P < 0.001), and 2.09 and 2.83 for males and females aged > or = 65 (Mantel-Haenszel weighted odds ratio = 2.19, P < 0.001). Male sex, age > or = 65, working at onset of illness, regular alcohol use, drug abuse and absence of contact history were associated with ever smokers (all P < 0.05). Ever smokers were more likely to have cough (OR 1.69), dyspnoea (OR 1.84), upper zone involvement (OR 1.67), cavity (OR 1.76), miliary lung involvement (OR 2.77), positive sputum culture (OR 1.43), but less isolated extrathoracic involvement (OR 0.31), even after controlling for the confounding background variables (all P < 0.05). CONCLUSION: There was a consistent association between smoking and tuberculosis. More aggressive lung involvement was also found among ever smokers. PMID- 14552571 TI - The assessment of IFN-gamma and its regulatory cytokines in the plasma and bronchoalveolar lavage fluid of patients with active pulmonary tuberculosis. AB - OBJECTIVES: To assess the role of IFN-gamma and its regulatory cytokines in active pulmonary tuberculosis (TB). DESIGN: Cytokines were measured in the plasma of TB patients and healthy subjects with different risk for TB exposure. In addition, cytokine profile was assessed in the bronchoalveolar lavage fluid (BALf) of six TB patients and nine normal controls. RESULTS: Circulating IFN gamma, IL-10 and IL-18 were higher in TB patients than in control groups. Plasma IL-12 levels were extremely variable, and no difference was observed among study groups. An inverse correlation between plasma IFN-gamma and IL-10 levels was found in TB patients. Furthermore, circulating IL-18 correlated with IL-10 but not with IFN-gamma levels. Finally, IFN-gamma, IL-18 and IL-12 were increased in the BALf of TB patients, whereas no difference was observed in IL-10 levels. CONCLUSIONS: In human TB, at least at certain disease stages, there is a differential compartmentalization of the IFN-gamma-regulatory factors IL-12 and IL-10, the former being concentrated in the lungs and the latter being present in peripheral circulation. In addition, our findings address more critically the role of IL-18 in the host response to tuberculosis infection in humans. PMID- 14552573 TI - Assessment of the Mycobacteria Growth Indicator Tube for the bacteriological diagnosis of tuberculosis. AB - Fast, accurate diagnosis is necessary for rapid treatment of patients and to prevent the spread of Mycobacterium tuberculosis strains. The rate of recovery, mean time to detection and contamination rates of the Mycobacteria Growth Indicator Tube (MGIT) were compared with Lowenstein-Jensen (LJ) medium for mycobacterial cultures performed on 405 clinical specimens decontaminated by the trisodium phosphate method without benzalkonium chloride. The recovery rate of M. tuberculosis using MGIT was 45/61 (73.8%) compared with the reference LJ. The mean times to detection of M. tuberculosis in smear-positive specimens were 11.9 days with MGIT and 20 days with LJ. For smear-negative samples, the mean times were respectively 18.6 and 31 days, and the contamination rates were respectively 4% and 1.2%. When the trisodium phosphate decontamination method is used, MGIT cannot be used alone for isolation of mycobacteria, but may be used in combination with LJ. PMID- 14552572 TI - Mycobacterium bovis (bovine TB) exposure as a recreational risk for hunters: results of a Michigan Hunter Survey, 2001. AB - SETTING: Tuberculosis caused by Mycobacterium bovis (bovine TB) is endemic in the white-tailed deer population of north-eastern Michigan. Hunters may be exposed to M. bovis via cutaneous inoculation while field dressing deer or by ingestion of undercooked venison. Michigan hunters have received inconsistent messages about their risk of acquiring tuberculosis from recreational exposure to deer. The most common health advice offered has been to wear gloves while field dressing deer and to cook venison products thoroughly. OBJECTIVE: Data were collected to quantify these self-protective activities and to characterize hunters practicing these activities. DESIGN: In 2001, we surveyed 1833 hunters who had successfully harvested deer in or near Michigan's bovine TB endemic area in 2000. RESULTS: The survey response rate was 78%. Most hunters (89%) reported field dressing deer, 43% of whom wore gloves. Most hunters (95%) reported eating venison, 55% of whom reported their venison was always cooked thoroughly. Several hunter characteristics, including older age, female sex, higher awareness level, and area of residence, were significantly associated with the practice of these self protective activities. CONCLUSION: The survey results suggest that hunters should receive consistent advice encouraging glove use while field dressing deer and the thorough cooking of venison products before consumption. PMID- 14552574 TI - Epidaure: a research project in medical image analysis, simulation, and robotics at INRIA. PMID- 14552575 TI - Cone-beam reprojection using projection-matrices. AB - This paper addresses reprojection of three-dimensional (3-D) reconstructions obtained from cone-beam scans using a C-arm imaging equipment assisted by a pose determining system. The emphasis is on reprojecting without decomposing the estimated projection matrix (P-matrix) associated with a pose. Both voxel- and ray-driven methods are considered. The voxel-driven reprojector follows the algorithm for backprojection using a P-matrix. The ray-driven reprojector is derived by extracting from the P-matrix the equation of the line joining a detector-pixel and the X-ray source position. This reprojector can be modified to a ray-driven backprojector. When the geometry is specified explicitly in terms of the physical parameters of the imaging system, the projection matrices can be constructed. The resulting "projection-matrix method" is advantageous, especially when the scanning trajectory is irregular. The algorithms presented are useful in iterative methods of image reconstruction and enhancement procedures, apart from their well-known role in visualization and volume rendering. Reprojections of 3-D patient data compare favorably with the original X-ray projections obtained from a prototype C-arm system. The algorithms for reprojection can be modified to compute perspective maximum intensity projection. PMID- 14552576 TI - A comparison of exact and approximate adjoint sensitivities in fluorescence tomography. AB - Many approaches to fluorescence tomography utilize some form of regularized nonlinear least-squares algorithm for data inversion, thus requiring repeated computation of the Jacobian sensitivity matrix relating changes in observable quantities, such as emission fluence, to changes in underlying optical parameters, such as fluorescence absorption. An exact adjoint formulation of these sensitivities comprises three terms, reflecting the individual contributions of 1) sensitivities of diffusion and decay coefficients at the emission wavelength, 2) sensitivities of diffusion and decay coefficients at the excitation wavelength, and 3) sensitivity of the emission source term. Simplifying linearity assumptions are computationally attractive in that they cause the first and second terms to drop out of the formulation. The relative importance of the three terms is thus explored in order to determine the extent to which these approximations introduce error. Computational experiments show that, while the third term of the sensitivity matrix has the largest magnitude, the second term becomes increasingly significant as target fluorophore concentration or volume increases. Image reconstructions from experimental data confirm that neglecting the second term results in overestimation of sensitivities and consequently overestimation of the value and volume of the fluorescent target, whereas contributions of the first term are so low that they are probably not worth the additional computational costs. PMID- 14552577 TI - Level-set-based artery-vein separation in blood pool agent CE-MR angiograms. AB - Blood pool agents (BPAs) for contrast-enhanced (CE) magnetic-resonance angiography (MRA) allow prolonged imaging times for higher contrast and resolution. Imaging is performed during the steady state when the contrast agent is distributed through the complete vascular system. However, simultaneous venous and arterial enhancement in this steady state hampers interpretation. In order to improve visualization of the arteries and veins from steady-state BPA data, a semiautomated method for artery-vein separation is presented. In this method, the central arterial axis and central venous axis are used as initializations for two surfaces that simultaneously evolve in order to capture the arterial and venous parts of the vasculature using the level-set framework. Since arteries and veins can be in close proximity of each other, leakage from the evolving arterial (venous) surface into the venous (arterial) part of the vasculature is inevitable. In these situations, voxels are labeled arterial or venous based on the arrival time of the respective surface. The evolution is steered by external forces related to feature images derived from the image data and by internal forces related to the geometry of the level sets. In this paper, the robustness and accuracy of three external forces (based on image intensity, image gradient, and vessel-enhancement filtering) and combinations of them are investigated and tested on seven patient datasets. To this end, results with the level-set-based segmentation are compared to the reference-standard manually obtained segmentations. Best results are achieved by applying a combination of intensity- and gradient-based forces and a smoothness constraint based on the curvature of the surface. By applying this combination to the seven datasets, it is shown that, with minimal user interaction, artery-vein separation for improved arterial and venous visualization in BPA CE-MRA can be achieved. PMID- 14552578 TI - Unsupervised robust nonparametric estimation of the hemodynamic response function for any fMRI experiment. AB - This paper deals with the estimation of the blood oxygen level-dependent response to a stimulus, as measured in functional magnetic resonance imaging (fMRI) data. A precise estimation is essential for a better understanding of cerebral activations. The most recent works have used a nonparametric framework for this estimation, considering each brain region as a system characterized by its impulse response, the so-called hemodynamic response function (HRF). However, the use of these techniques has remained limited since they are not well-adapted to real fMRI data. Here, we develop a threefold extension to previous works. We consider asynchronous event-related paradigms, account for different trial types and integrate several fMRI sessions into the estimation. These generalizations are simultaneously addressed through a badly conditioned observation model. Bayesian formalism is used to model temporal prior information of the underlying physiological process of the brain hemodynamic response. By this way, the HRF estimate results from a tradeoff between information brought by the data and by our prior knowledge. This tradeoff is modeled with hyperparameters that are set to the maximum-likelihood estimate using an expectation conditional maximization algorithm. The proposed unsupervised approach is validated on both synthetic and real fMRI data, the latter originating from a speech perception experiment. PMID- 14552579 TI - Three-dimensional guide-wire reconstruction from biplane image sequences for integrated display in 3-D vasculature. AB - Using three-dimensional rotational X-ray angiography (3DRA), three-dimensional (3 D) information of the vasculature can be obtained prior to endovascular interventions. However, during interventions, the radiologist has to rely on fluoroscopy images to manipulate the guide wire. In order to take full advantage of the 3-D information from 3DRA data during endovascular interventions, a method is presented that yields an integrated display of the position of the guide wire and vasculature in 3-D. The method relies on an automated method that tracks the guide wire simultaneously in biplane fluoroscopy images. Based on the calibrated geometry of the C-arm, the 3-D guide-wire position is determined and visualized in the 3-D coordinate system of the vasculature. The method is evaluated in an intracranial anthropomorphic vascular phantom. The influence of the angle between projections, distortion correction of the projection images, and accuracy of geometry knowledge on the accuracy of 3-D guide-wire reconstruction from biplane images is determined. If the calibrated geometry information is used and the images are corrected for distortion, a mean distance to the reference standard of 0.42 mm and a tip distance of 0.65 mm is found, which means that accurate guide wire reconstruction from biplane images can be performed. PMID- 14552580 TI - Three-dimensional segmentation and growth-rate estimation of small pulmonary nodules in helical CT images. AB - Small pulmonary nodules are a common radiographic finding that presents an important diagnostic challenge in contemporary medicine. While pulmonary nodules are the major radiographic indicator of lung cancer, they may also be signs of a variety of benign conditions. Measurement of nodule growth rate over time has been shown to be the most promising tool in distinguishing malignant from nonmalignant pulmonary nodules. In this paper, we describe three-dimensional (3 D) methods for the segmentation, analysis, and characterization of small pulmonary nodules imaged using computed tomography (CT). Methods for the isotropic resampling of anisotropic CT data are discussed. 3-D intensity and morphology-based segmentation algorithms are discussed for several classes of nodules. New models and methods for volumetric growth characterization based on longitudinal CT studies are developed. The results of segmentation and growth characterization methods based on in vivo studies are described. The methods presented are promising in their ability to distinguish malignant from nonmalignant pulmonary nodules and represent the first such system in clinical use. PMID- 14552581 TI - Three-dimensional edge-preserving image enhancement for computed tomography. AB - Computed tomography (CT) images exhibit a variable amount of noise and blur, depending on the physical characteristics of the apparatus and the selected reconstruction method. Standard algorithms tend to favor reconstruction speed over resolution, thereby jeopardizing applications where accuracy is critical. In this paper, we propose to enhance CT images by applying half-quadratic edge preserving image restoration (or deconvolution) to them. This approach may be used with virtually any CT scanner, provided the overall point-spread function can be roughly estimated. In image restoration, Markov random fields (MRFs) have proven to be very flexible a priori models and to yield impressive results with edge-preserving penalization, but their implementation in clinical routine is limited because they are often viewed as complex and time consuming. For these practical reasons, we focused on numerical efficiency and developed a fast implementation based on a simple three-dimensional MRF model with convex edge preserving potentials. The resulting restoration method provides good recovery of sharp discontinuities while using convex duality principles yields fairly simple implementation of the optimization. Further reduction of the computational load can be achieved if the point-spread function is assumed to be separable. Synthetic and real data experiments indicate that the method provides significant improvements over standard reconstruction techniques and compares well with convex-potential Markov-based reconstruction, while being more flexible and numerically efficient. PMID- 14552582 TI - Region-based wavelet coding methods for digital mammography. AB - Spatial resolution and contrast sensitivity requirements for some types of medical image techniques, including mammography, delay the implementation of new digital technologies, namely, computer-aided diagnosis, picture archiving and communications systems, or teleradiology. In order to reduce transmission time and storage cost, an efficient data-compression scheme to reduce digital data without significant degradation of medical image quality is needed. In this study, we have applied two region-based compression methods to digital mammograms. In both methods, after segmenting the breast region, a region-based discrete wavelet transform is applied, followed by an object-based extension of the set partitioning in hierarchical trees (OB-SPIHT) coding algorithm in one method, and an object-based extension of the set partitioned embedded block (OB SPECK) coding algorithm in the other. We have compared these specific implementations against the original SPIHT and the new standard JPEG 2000, both using reversible and irreversible filters, on five digital mammograms compressed at rates ranging from 0.1 to 1.0 bit per pixel (bbp). Distortion was evaluated for all images and compression rates by the peak signal-to-noise ratio. For all images, OB-SPIHT and OB-SPECK performed substantially better than the traditional SPIHT and JPEG 2000, and a slight difference in performance was found between them. A comparison applying SPIHT and the standard JPEG 2000 to the same set of images with the background pixels fixed to zero was also carried out, obtaining similar implementation as region-based methods. For digital mammography, region based compression methods represent an improvement in compression efficiency from full-image methods, also providing the possibility of encoding multiple regions of interest independently. PMID- 14552583 TI - The perception of breast cancers--a spatial frequency analysis of what differentiates missed from reported cancers. AB - The primary detector of breast cancer is the human eye. Radiologists read mammograms by mapping exogenous and endogenous factors, which are based on the image and observer, respectively, into observer-based decisions. These decisions rely on an internal schema that contains a representation of possible malignant and benign findings. Thus, to understand the hits and misses made by the radiologists, it is important to model the interactions between the measurable image-based elements contained in the mammogram and the decisions made. The image based elements can be of two types, i.e., areas that attracted the visual attention of the radiologist, but did not yield a report, and areas where the radiologist indicated the presence of an abnormal finding. In this way, overt and covert decisions are made when reading a mammogram. In order to model this decision-making process, we use a system that is based upon the processing done by the human visual system, which decomposes the areas under scrutiny in elements of different sizes and orientations. In our system, this decomposition is done using wavelet packets (WPs). Nonlinear features are then extracted from the WP coefficients, and an artificial neural network is trained to recognize the patterns of decisions made by each radiologist. Afterwards, the system is used to predict how the radiologist will respond to visually selected areas in new mammogram cases. PMID- 14552584 TI - Heart-surface reconstruction and ECG electrodes localization using fluoroscopy, epipolar geometry and stereovision: application to noninvasive imaging of cardiac electrical activity. AB - To date there is no imaging modality for cardiac arrhythmias which remain the leading cause of sudden death in the United States (> 300000/yr.). Electrocardiographic imaging (ECGI), a noninvasive modality that images cardiac arrhythmias from body surface potentials, requires the geometrical relationship between the heart surface and the positions of body surface ECG electrodes. A photographic method was validated in a mannequin and used to determine the three dimensional coordinates of body surface ECG electrodes to within 1 mm of their actual positions. Since fluoroscopy is available in the cardiac electrophysiology (EP) laboratory where diagnosis and treatment of cardiac arrhythmias is conducted, a fluoroscopic method to determine the heart surface geometry was developed based on projective geometry, epipolar geometry, point reconstruction, b-spline interpolation and visualization. Fluoroscopy-reconstructed hearts in a phantom and a human subject were validated using high-resolution computed tomography (CT) imaging. The mean absolute distance error for the fluoroscopy reconstructed heart relative to the CT heart was 4 mm (phantom) and 10 mm (human). In the human, ECGI images of normal cardiac electrical activity on the fluoroscopy-reconstructed heart showed close correlation with those obtained on the CT heart. Results demonstrate the feasibility of this approach for clinical noninvasive imaging of cardiac arrhythmias in the interventional EP laboratory. PMID- 14552585 TI - FAME--a flexible appearance modeling environment. AB - Combined modeling of pixel intensities and shape has proven to be a very robust and widely applicable approach to interpret images. As such the active appearance model (AAM) framework has been applied to a wide variety of problems within medical image analysis. This paper summarizes AAM applications within medicine and describes a public domain implementation, namely the flexible appearance modeling environment (FAME). We give guidelines for the use of this research platform, and show that the optimization techniques used renders it applicable to interactive medical applications. To increase performance and make models generalize better, we apply parallel analysis to obtain automatic and objective model truncation. Further, two different AAM training methods are compared along with a reference case study carried out on cross-sectional short-axis cardiac magnetic resonance images and face images. Source code and annotated data sets needed to reproduce the results are put in the public domain for further investigation. PMID- 14552586 TI - An Eulerian PDE approach for computing tissue thickness. AB - We outline an Eulerian framework for computing the thickness of tissues between two simply connected boundaries that does not require landmark points or parameterizations of either boundary. Thickness is defined as the length of correspondence trajectories, which run from one tissue boundary to the other, and which follow a smooth vector field constructed in the region between the boundaries. A pair of partial differential equations (PDEs) that are guided by this vector field are then solved over this region, and the sum of their solutions yields the thickness of the tissue region. Unlike other approaches, this approach does not require explicit construction of any correspondence trajectories. An efficient, stable, and computationally fast solution to these PDEs is found by careful selection of finite differences according to an upwinding condition. The behavior and performance of our method is demonstrated on two simulations and two magnetic resonance imaging data sets in two and three dimensions. These experiments reveal very good performance and show strong potential for application in tissue thickness visualization and quantification. PMID- 14552587 TI - Modeling the interaction of paclitaxel with beta-tubulin. AB - The natural product cytotoxic agent, paclitaxel, partly induces cell death through its ability to disrupt mitosis by binding to the microtubule protein beta tubulin. Structural characterization of the paclitaxel-beta-tubulin complex, a first stage in the design of new antimitotics, has been complicated by contradictory observations obtained from different experimental techniques [electron crystallography, fluorescence resonance energy transfer (FRET), and photo-affinity labeling (PAL)] used to examine the complex. In this study we have used a range of molecular modeling techniques including restrained conformational searching and computer-assisted docking to propose that these contradictions may be resolved by the hypothesis that the nature of the interaction of paclitaxel with beta-tubulin depends on the physical form of the tubulin examined. In particular, our analysis identified a binding mode that is consistent with available data for the interaction of paclitaxel with beta-tubulin in polymerized tubulin or microtubules. This orientation is characterized by an alternate conformation (inverted orientation of side chains) and inverted orientation of the taxane core of paclitaxel within its tubulin binding site compared with the electron crystallographic structure. The proposed structure, however, is only marginally consistent with electron crystallographic data for the interaction of paclitaxel with beta-tubulin in Zn-induced tubulin sheets. Similarly, the electron crystallographic structure shows poor correlation with FRET, solid-state NMR, and some observed SAR relationships for paclitaxel interacting with polymerized tubulin or microtubules. These observations suggest to us that the interaction of paclitaxel with Zn-induced tubulin sheets may not reflect paclitaxel's interaction with tubulin in microtubules and hence may not be an appropriate guide for rational drug design programs. PMID- 14552588 TI - Reductive metabolism of the nitroimidazole-based hypoxia-selective cytotoxin NLCQ 1 (NSC 709257). AB - The enzymatic cell-free metabolism of the novel hypoxia-selective cytotoxin 4-[3 (2-nitro-1-imidazolyl)-propylamino]-7-chloroquinoline hydrochloride (NLCQ-1) was investigated under hypoxic or aerobic conditions in the presence of purified reductive enzymes or isolated rat liver microsomes by monitoring the parent compound with HPLC-UV analysis. Enzymatic reduction of NLCQ-1 with isolated rat liver microsomes and NADPH or NADH showed that, only under hypoxic conditions, ca. 45% and 60% of the parent compound was reduced, respectively, within 1 h of incubation (37 degrees C). Under identical conditions but in the presence of 2' AMP (a P450 reductase inhibitor), 6-propyl-2-thiouracil or p hydroxymercuribenzoate (cytochrome b5 reductase inhibitors), NLCQ-1 reduction was inhibited. Enzymatic cell-free metabolism of NLCQ-1 with recombinant human DT diaphorase (DTD) and NADPH or NADH under hypoxic or aerobic conditions showed that < or = 5% of the compound was reduced within 2 h. Reduction kinetics with human P450 reductase-expressing microsomes showed ca. 75% or 50% reduction of NLCQ-1 under hypoxic or aerobic conditions, respectively, after 2 h incubation. These results suggest that DTD is not involved in the initial steps of the bioreductive metabolism of NLCQ-1, although it could be involved with metabolites of NLCQ-1, and that cytochrome P450 and cytochrome b5 reductases play a significant role in the bioreductive metabolism of NLCQ-1. PMID- 14552589 TI - Increased activity and expression of phospholipase D2 in human colorectal cancer. AB - Phospholipase D (PLD) has been reported as relevant to some types of human cancer, but its role in human colorectal cancer still remains to be elucidated. Thus, this study was conducted to determine the activity and the expression of PLD2 in human colorectal cancer. A significant elevation of PLD2 activity and higher expression of PLD2 protein were detected in human colorectal cancer in comparison with corresponding normal mucosa. The tendency of higher expression of PLD2 mRNA was also observed. The ratio of PLD2 activity in cancer to that in corresponding normal mucosa was greater in colorectal cancer with nodal involvement and deeper tumor invasion. Our results indicate that PLD2 has a possible implication in carcinogenesis and progression and would be a new therapeutic target and a potential tumor marker for colorectal cancer. PMID- 14552590 TI - Binding site(s) on P-glycoprotein for a newly synthesized photoaffinity analog of agosterol A. AB - Agosterol A (AG-A) is a novel agent that reverses P-glycoprotein (P-gp) and multidrug resistance protein-1 (MRP1)-meditated multidrug resistance (MDR). We have synthesized [125I]11-azidophenyl agosterol A (azidoAG-A), a photoaffinity analog of AG-A, and characterized its binding to P-gp in membrane vesicles prepared from multidrug-resistant P-gp-overexpressing KB-C2 cells. The photoanalog photolabeled intact P-gp and both the N- and C-terminal fragments of P-gp. [125I]AzidoAG-A is transported by P-gp and the intracellular accumulation of both [125I]azidoAG-A and [3H]AG-A in KB-C2 cells was lower than that in the parental drug-sensitive KB-3-1 cells. [125I]AzidoAG-A bound to the drug binding site(s) on P-gp because photoaffinity labeling of P-gp was inhibited by a variety of known P-gp substrates, including anticancer, reversing, and anti-human immunodeficiency virus (HIV) agents. The binding of [125I]azidoAG-A to P-gp differs from the binding of other photolabeled probes such as iodoaryl azidoprazosin (IAAP) to P-gp and from the binding of [125I]azidoAG-A to MRP1 based on the differing effects of flupentixol and glutathione (GSH) on their binding. Thus, [125I]azidoAG-A will be a useful tool to elucidate the structure and function of P-gp because it directly binds to the drug binding site(s) on P gp, is transported by P-gp, and exhibits different P-gp binding characteristics than IAAP. PMID- 14552591 TI - Development of a syngeneic in vivo tumor model and its use in evaluating a novel P-glycoprotein modulator, PGP-4008. AB - Multidrug resistance (MDR) is a phenomenon by which tumor cells develop reduced sensitivity to anticancer drugs, which often leads to the failure of cancer chemotherapy. A prominent mechanism of MDR is the overexpression of the multidrug efflux pump, P-glycoprotein (P-gp), that decreases the intracellular accumulation of many anticancer drugs, leading to increased tumor growth. Intensive efforts are under way to develop clinically useful MDR modulators that inhibit the function of P-gp for use in combination with established anticancer drugs. Our goal was to develop an improved in vivo solid tumor model utilizing immunocompetent animals to examine the efficacy of P-gp-specific MDR modulators. Using in vitro cytotoxicity and drug accumulation assays, two transformed murine cell lines, JC and TIB-75, were found to demonstrate the P-gp-mediated MDR phenotype. In contrast, two similar lines did not express functional P-gp. Western blot analyses confirmed the expression of P-gp and the lack of expression of the closely related drug efflux protein MRP1 in the JC and TIB-75 cell lines. The JC cell line displayed excellent tumorigenicity and consistent growth kinetics when implanted into immune-competent Balb/c mice. Animals treated with a combination of a known MDR modulator, cyclosporin A, and a cytotoxic drug, doxorubicin, exhibited significantly reduced tumor growth compared with untreated controls or animals treated with either cyclosporin A or doxorubicin alone. Similarly, a novel P-gp-specific MDR modulator, PGP-4008, in combination with doxorubicin showed inhibition of tumor growth. However, in contrast with the significant loss of body weight observed in the animals treated with the combination of cyclosporin A and doxorubicin, those treated with PGP-4008 plus doxorubicin did not experience weight loss. Therefore, this syngeneic solid tumor model provides a new in vivo system that can be used to evaluate the efficacy of P-gp inhibitors in an immune-competent host. This should allow improved prediction of the clinical utility of these compounds. PMID- 14552592 TI - Filamentary keratopathy: a non-contact photomicrographic in vivo study in the human cornea. AB - PURPOSE: To investigate in vivo morphology of corneal filaments. MATERIALS AND METHODS: Over a 12-year period, 19 patients with corneal filaments (7 with keratoconjunctivitis sicca and 12 with other surface diseases) were examined with the slit lamp and photographed by non-contact photomicrography. RESULTS: The filaments appeared as variously long and thick threads, attached at one or both ends to the surface and carrying various amounts of granular and amorphous material. The attachment points and their surroundings showed abnormal cells and diffusion of tear fluid stained green with fluorescein sodium. The underlying stroma appeared normal. The remaining epithelium often showed abnormal surface cells and/or edematous or cystic changes. CONCLUSIONS: The in vivo morphology of filamentary keratopathy is consistent with aggregations of mucus and cell debris adhering to the corneal surface. The underlying diseases seem to have in common corneal epithelial edema. The nature of additional factors, or their combination, precipitating mucus adherence to the surface is not clear. PMID- 14552593 TI - Tumor necrosis factor-alpha (TNF-alpha) in seasonal allergic conjunctivitis and vernal keratoconjunctivitis. AB - PURPOSE: To quantify the presence of the proinflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) in allergic conjunctivitis. MATERIALS AND METHODS: Tears and peripheral blood samples were collected from patients with seasonal allergic conjunctivitis (SAC, n=6), vernal keratoconjunctivitis (VKC, n=12), and normal subjects (CT, n=12). From an additional six nonactive allergic patients, tears were collected before and after specific conjunctival allergen challenge (CAC). Upper tarsal conjunctival biopsies were obtained from five CT and five VKC patients. TNF-alpha in tears was measured by enzyme-linked immunoassay and identified in tissues by immunohistochemistry. RESULTS: Tear TNF-alpha levels in VKC patients were significantly increased compared to CT (p=0.03), and were significantly correlated with the severity of the disease. No differences were found between SAC and CT tear samples. TNF-alpha serum levels were higher in VKC than CT, however, this difference was not statistically significant. After CAC, tear TNF-alpha levels were found increased in only one of six patients. In VKC tissues, TNF-alpha positive cells were significantly increased compared to CT (p=0.03). CONCLUSIONS: TNF-alpha may have a significant role in severe forms of allergic conjunctivitis. PMID- 14552594 TI - Effect of brimonidine on intraocular pressure in normal tension glaucoma: a short term clinical trial. AB - PURPOSE: To evaluate the efficacy and safety of 0.2% brimonidine eye drops given twice daily in normal tension glaucoma. PATIENTS AND METHODS: Sixteen consecutive patients fulfilling eligibility criteria (glaucomatous optic neuropathy associated with visual field defect in at least one eye, intraocular pressure (IOP) < or = 18 mmHg (average of the two highest readings of the round-the-clock curve, including one reading at midnight in supine position), no prior glaucoma therapy, angle wide open, visual acuity 20/40 or better) were enrolled in this prospective, randomized, placebo-controlled clinical trial with crossover design, lasting 30 days for each treatment phase plus 15-day washout in between. Main outcome was IOP (average of the two highest readings of the round-the-clock curve). RESULTS: Mean IOP was significantly reduced by brimonidine (from 17.1 +/- 0.7 mm Hg to 13.9 +/- 2.2 mmHg, p<0.001 (paired Student t-test)). At the end of the 30-day brimonidine phase, 4 of 16 subjects showed a > or = 30% IOP decrease over baseline. CONCLUSIONS: In the short term, 0.2% brimonidine eye drops can induce a significant IOP decrease in eyes with normal tension glaucoma. PMID- 14552595 TI - Prognosis of juvenile rheumatoid arthritis-associated uveitis. AB - PURPOSE: To evaluate the clinical characteristics and the visual prognosis of uveitis in juvenile rheumatoid arthritis (JRA). METHODS: The authors examined 63 patients with uveitis and JRA observed from January 1985 to December 2000. The following characteristics of each patient were considered: age at first visit, age at onset of uveitis and arthritis, sex, laterality and localization of uveitis, ocular complications, antinuclear antibody (ANA) and human leukocyte antigen (HLA) DR11 positivity, and follow-up. A retrospective study on mid-time visual outcome and ocular complications was performed on 42 patients with more than 12 months of follow-up. RESULTS: A total of 76.2% of the patients were female, with a mean age of 8.1 years. Chronic anterior uveitis was bilateral in 77.8% of the cases and unilateral in 22.2%. Arthritis was oligoarticular at onset in 87.3% of cases, and polyarticular in 12.7%. Mean age at arthritis onset was 4.5 years and mean age at uveitis onset was 5.4 years. ANA were positive in 92% of cases and HLA DR11 was present in 36 of the 43 patients tested (83.7%). Among the 42 patients with more than 12 months of follow-up, ocular complications occurred in 90.5% of cases and the most frequent were cataract (64.4% of eyes) and band keratopathy (59.2% of eyes). Secondary glaucoma (25% of eyes) was associated with the worst visual prognosis. A total of 64.5% of eyes maintained a visual acuity between 20/33 and 20/20 at the end of the follow-up. CONCLUSIONS: Visual prognosis of uveitis associated with JRA is improving, owing to earlier diagnosis and intensive treatment. Ocular complications occurred frequently in patients with uveitis and JRA but they did not seem to seriously affect the final visual outcome. The authors did not observe any correlation between prognosis and sex, age at the onset of uveitis or arthritis, pattern of arthritis, or positivity for ANA or HLA DR11. In a percentage of cases, uveitis may develop before arthritis or years after the onset of arthritis; therefore, continuous ophthalmologic examinations are needed in young people with JRA. PMID- 14552596 TI - Intraoperative biometry for intraocular lens (IOL) power calculation at silicone oil removal. AB - PURPOSE: Cataract development is common following retinal detachment surgery that necessitates silicone oil injection. Intraocular lens (IOL) power calculation in the presence of silicone oil is challenging for many reasons. The authors evaluated the accuracy of intraoperative biometry during cataract surgery in silicone-filled eyes. METHODS: Twelve cases of cataract in eyes filled with silicone oil after retinal detachment surgery were included. Preoperatively, keratometric readings were documented. Intraoperative axial length was measured following removal of silicone oil using a sterile probe of the Nidek Echoscan US800 unit. IOL power calculation was computed using the SRK/T formula. This was followed by phacoemulsification and foldable lens implantation. Postoperative refraction allowed evaluation of the accuracy of intraoperative biometry. The predictability of three different formulas was also compared (Holladay, SRK/T, and SRK II). RESULTS: Seven men and five women with a mean age of 50.2 years were included in the study. The mean postoperative spherical equivalent using the described technique was 6.77 +/- 0.43 diopters. SRK/T and Holladay formula had the best predictability for high axial length eyes, whereas the three studied formulas showed no significant differences in predictability in eyes with normal axial length. CONCLUSIONS: Intraoperative biometry proved to have good predictability for the absolute postoperative refractive error in cataract surgery for eyes at the time of silicone oil removal. This predictability was accurate independent of axial length. PMID- 14552597 TI - Secondary intraocular lens (IOL) implantation: anterior chamber versus scleral fixation long-term comparative evaluation. AB - PURPOSE: To compare the long-term efficacy of a secondarily implanted flexible angle supported anterior chamber (AC) intraocular lens (IOL) with sclerally fixated IOL as a secondary procedure in patients with aphakia. METHODS: Sixty patients with aphakia were recruited for this study. The patients were randomly divided into two groups: Group I patients underwent secondary implantation with AC IOL (Kelman Z type multiflex Domi classic AC IOL) and Group II patients underwent secondary implantation with scleral fixated IOL (Hanita lens with two eyelets). Postoperatively, the patients were followed up at 1 week, monthly for 6 months, and at twice yearly intervals for 5 years (mean follow-up was 3 years). RESULTS: Best-corrected visual acuity of 6/18 or better was achieved in 36.6% (11/30) of patients in Group I and 30% (9/30) of patients in Group II after a mean follow-up of 3 years. The complication rate was higher in Group II as compared to Group I. The authors encountered a total of 23 complications in the sclerally fixated group and 11 in the AC IOL group (p=0.004). Uveitis and ciliary tenderness were the most common complications in Groups I and II, respectively. CONCLUSIONS: For secondary implantation of IOL in aphakic patients, Kelman Z type multiflex Domi classic AC IOL is a better rehabilitation modality as compared to the scleral fixated Hanita IOL. PMID- 14552598 TI - Multinucleated giant cells in periretinal silicone granulomas are associated with progressive proliferative vitreoretinopathy. AB - PURPOSE: To determine the histologic features of granulomatous reactions in persilicone periretinal proliferation. PATIENTS AND METHODS: This retrospective study included 12 patients with recurrent retinal detachment and persilicone granulomatous proliferation after vitrectomy for proliferative vitreoretinopathy (PVR). All patients underwent reoperation for membrane surgery. Immunohistochemical study of the excised periretinal membranes was performed with cytokeratins, GFAP, vimentin, CD68, CD45, and lysozyme antibodies. RESULTS: The cellular characteristics of periretinal granulomas allow differentiation of two types of tissue. Spongy tissue (nine cases) showed an accumulation of mature vacuolated macrophages that contained silicone without multinucleated giant cells (MGC). The second type (three cases) consisted of an accumulation of sparsely vacuolated macrophages, epithelioid cells, and MGC. The MGC corresponded to transition forms of foreign body giant cells (FBGC). Spongy tissue was associated with anatomic success (58.3% of cases) and with stabilized PVR (66.7% of cases) at the time of the membrane surgery. MGC were associated with prolonged silicone oil tamponade, recurrent retinal detachment, and progressive PVR. CONCLUSIONS: Intraocular silicone oil can lead to periretinal foreign body granulomas. FBGC are occasionally observed and were associated with progressive PVR. PMID- 14552599 TI - Aprotinin reduces ischemia-reperfusion injury in the retina of guinea pigs. AB - PURPOSE: The aim of this study was investigate the role of aprotinin on retinal lipid peroxidation and histopathological changes during ischemia/reperfusion (I/R) of guinea pigs. METHODS: Three groups of seven pigmented guinea pigs each were formed: a control (group 1), ischemia/saline (group 2) and ischemia/aprotinin (group 3). One eye of each animal was selected for histopathological evaluation and the other for biochemical assay. Bilateral pressure-induced retinal ischemia was instigated for 90 min and was followed by 24 hours of reperfusion. Animals in the ischemia/aprotinin and ischemia/saline groups received either 20,000 KIU/kg of aprotinin or saline, repeated four times at 6-hour intervals, with the first dose administered 5 min prior to the ischemic insult. The animals were killed at 24 hours of reperfusion. Retinal malondialdehyde (MDA) levels and the thickness of the inner plexiform layers were measured. RESULTS: The level of MDA in group 1 was significantly (p<0.001) lower than the other groups. The mean MDA level in group 2 was significantly (p<0.01) higher than in group 3. The inner plexiform layer in group 1 was significantly (p<0.001) thinner than in the other groups. The mean thickness of the inner plexiform layer in group 2 was significantly (p<0.01) higher than in group 3. CONCLUSIONS: These data indicate that intraperitoneally administrated aprotinin has a protective effect against I/R injury in the retina of guinea pig as evidenced by reduced retinal MDA level and retinal thickness. PMID- 14552601 TI - Free-floating cyst in the anterior chamber: ultrasound biomicroscopic reports. AB - PURPOSE: To report a free-floating dislodged pigmented cyst in the anterior chamber and to describe the ultrasound biomicroscopic (UBM) features of the lesion. METHODS: Case report. RESULTS: A 26-year-old man presented with noise in his left eye in the prone position. Slit-lamp examination revealed a small pigmented mass lesion at the 6 o'clock position in the anterior chamber, slowly moving with changing head postures. UBM revealed an unfixed dislodged pigmented cyst with a thin wall and no internal reflectivity, changing its shape from ovoid to circular. Five years later, the anterior segment does not show any modification and the cyst has not changed in size or appearance. CONCLUSIONS: Cysts of the iris pigment epithelium arise in the posterior iris layer. They may be central, midzonal, peripheral, or dislodged. Dislodged cysts may be repositioned and fixed or free-floating in the vitreous or in the anterior chamber. Surgical removal must be considered only for a rapid enlargement or significant reduction in endothelial cell count. UBM can distinguish solid from cystic lesions, giving detailed information on internal structure, reflectivity, shape, and thickness. PMID- 14552600 TI - Diabetes mellitus: a risk factor affecting visual outcome in branch retinal vein occlusion. AB - PURPOSE: The prognosis of visual acuity (VA) after branch retinal vein occlusion (BRVO) in patients with diabetes mellitus is unknown compared to the VA in non diabetic patients with BRVO. The aim of this study was to evaluate the visual outcome of BRVO in diabetic and non-diabetic patients. METHODS: A retrospective case-control study of diabetic and non-diabetic patients with BRVO was performed. VA and commonly known risk factors and complications of BRVO were compared in a follow-up period of at least 1 year. RESULTS: A total of 28 eyes of patients with diabetes and 49 eyes of non-diabetic patients with BRVO were included. One year after BRVO, the VA in the patients with diabetes decreased significantly more than that of the non-diabetic patients. During the second year after BRVO, the VA did not change significantly in either group. BRVO in patients with diabetes occurs at an earlier age. Diabetic patients needed more outpatient visits. CONCLUSIONS: The VA 1 year after BRVO in patients with diabetes is worse compared to the VA in patients without diabetes. The VA stabilizes 1 year after onset in both groups. Diabetic patients tend to need more frequent follow-up in order to treat the sequelae of BRVO. PMID- 14552602 TI - Choroidal hemangioma treated with photodynamic therapy using verteporfin: report of a case. AB - PURPOSE: To report the results of treatment of circumscribed choroidal hemangioma with a single application of photodynamic therapy (PDT) with verteporfin according to the Treatment of Age-related Macular Degeneration with Photodynamic Therapy study. METHODS: A 44-year-old man with unilateral decreased vision and macular subretinal fluid secondary to a circumscribed choroidal hemangioma diagnosed by fluorescein and indocyanine green angiography and ultrasonography underwent PDT with verteporfin therapy. RESULTS: One year after PDT, subretinal fluid was absent and visual acuity improved. CONCLUSIONS: The results obtained in this case are in keeping with previously reported results; however, future randomized studies are necessary to evaluate and standardize different infusion times in order to obtain maximum efficacy of treatment. PMID- 14552603 TI - Herpes simplex virus acute retinal necrosis during pregnancy. AB - PURPOSE: As pregnancy is liable to modify immune response, the authors explored the immune functions of a pregnant patient with acute retinal necrosis (ARN) to ascertain whether pregnancy may promote the onset of infection. METHODS: Polymerase chain reaction (PCR) was used for the detection of herpes simplex virus (HSV) DNA in ocular, uterus cervix, and cerebrospinal fluid samples. Peripheral blood mononuclear cells were cultured for 72 hours with mitogens and cellular proliferation was assessed using (methyl-3H) thymidine incorporation. Flow cytometry was performed for T, B, and NK cell count using CD2, CD3, CD4, CD8 (T cells), CD19, CD20 (B cells), and a combination of CD3-CD16 and CD56 monoclonal antibodies (NK cells). RESULTS: Unilateral ARN, with a confluent peripheral necrotizing retinitis extending throughout the entire retina, was diagnosed clinically. The herpetic infection (herpes simplex virus 1) was confirmed using PCR of aqueous humor specimen. The immunologic study performed during and after pregnancy showed that T and B lymphocytes were quantitatively normal and responses to concanavalin A, phytohemagglutinin, and pokeweed mitogens were weaker during pregnancy. CONCLUSIONS: A reduced response to mitogens, with postdelivery normalization, was noted in a pregnant woman with an ARN syndrome. Further studies are needed to explore the antigen-specific immune deviation in pregnant patients with ARN. PMID- 14552604 TI - Kinetic stabilization of primary hydrides of main group elements. The synthesis of an air-stable, crystalline arsine and silane. AB - Two new, "user-friendly" derivatives of triptycene containing AsH(2) and SiH(3) fragments were synthesized. Both solids are crystalline, air-stable compounds characterized by elevated melting points and resistance toward moisture. The highly reactive As-H and Si-H bonds are protected by the presence of the surrounding phenylene hydrogen atoms, which ensure a remarkable kinetic stabilization of these primary hydrides. After X-ray irradiation of a single crystal of triptycenesilane, a persistent silyl radical was trapped and characterized. PMID- 14552605 TI - [CH(3)(CH(2))(11)NH(3)]SnI(3): a hybrid semiconductor with MoO(3)-type tin(II) iodide layers. AB - The organic-inorganic hybrid [CH(3)(CH(2))(11)NH(3)]SnI(3) presents a lamellar structure with a Sn-I framework isotypic to that of MoO(3). The SnI(3)(-) layer consists of edge and corner-sharing SnI(6) octahedra in which one of the six Sn-I bonds is distinctly elongated (e.g., 3.62 A), indicating lone-pair stereoactivity for the Sn(II) atom. The overall electronic character remains comparable with that of the well-studied SnI(4)(2)(-)-based perovskite semiconductors, such as [CH(3)(CH(2))(11)NH(3)](2)SnI(4), with a red-shifted and broadened exciton peak associated with the band gap, apparently due to the increased dimensionality of the Sn-I framework. The title compound offers, aside from the hybrid perovskites, a new type of solution-processable Sn-I network for potential applications in semiconductive devices. PMID- 14552606 TI - Titanium and zirconium amido complexes ligated by 2,2'-Di(3 methylindolyl)methanes: synthesis, characterization, and ethylene polymerization activity. AB - 2,2'-Di(3-methylindolyl)methanes (L(2)H(2)) are introduced as dianionic, bidentate ligands of reduced pi-donating ability. Four complexes of the type L(2)Ti(NEt(2))(2) and L(2)Zr(NEt(2))(2)(THF) have been synthesized and characterized by elemental analysis, NMR ((1)H, (13)C) spectroscopy, and X-ray crystallography. Structural data confirm the reduced pi-donating ability of the eta(1)-indolyl moiety compared to that of diethylamido. Preliminary catalytic activities of these group 4 complexes for the polymerization of ethylene are reported. PMID- 14552607 TI - [Ge(7)O(13)(OH)(2)F(3)](3)(-).Cl(-).2[Ni(dien)(2)](2+): the first chainlike germanate templated by a transition metal complex. AB - The first chainlike germanate, [Ge(7)O(13)(OH)(2)F(3)](3)(-).Cl( ).2[Ni(dien)(2)](2+), has been solvothermally synthesized by using Ni(dien)(2)(2+) cations as the template and characterized by IR, SEM, TGA, powder X-ray diffraction (PXRD), energy-dispersive X-ray analysis (EDXA), elemental analysis, and single-crystal X-ray diffraction, respectively. This compound crystallized in the monoclinic space group P2/nwith a = 8.8904(2) A, b = 17.4374(3) A, c = 13.2110(3) A, beta = 101.352(1) degrees, V = 2007.97(7) A(3), and Z = 2. Interestingly, the structure contains two types of chiral mer [Ni(dien)(2)](2+) cations and two types of chiral chains, one left-handed and the other right-handed, which lead to a racemic compound. The orderly separation of achiral s-fac-[Ni(dien)(2)](2+) and chiral mer-[Ni(dien)(2)](2+) isomers was found in the structure. The structure is stabilized by N-H.O(F, Cl) hydrogen bonds. PMID- 14552609 TI - A novel undecametallic iron(III) cluster with an S = (11)/(2) spin ground state. AB - The reaction of [NEt(4)](2)[Fe(2)OCl(6)] with sodium benzoate, 4,6-dimethyl-2 hydroxypyrimidine (dmhp), and 1,1,1-tris(hydroxymethyl)ethane (H(3)thme) gives the undecametallic compound [NEt(4)][Fe(11)O(4)(O(2)CPh)(10)(thme)(4)(dmhp)(2)Cl(4)]. X-ray crystallography, EPR spectroscopy, bulk magnetic susceptibility studies, and low-temperature single-crystal magnetic measurements were used to characterize the compound. Magnetic measurements indicate an S = (11)/(2) ground state with the parameters g = 2.03 and D = -0.46 cm(-)(1). Single-crystal magnetic studies show hysteresis of molecular origin at T < 1.2 K with fast quantum mechanical tunneling at zero field. PMID- 14552608 TI - Low-potential cyclometalated osmium(II) mediators of glucose oxidase. AB - The osma(II)cycles [Os(phpy)(LL)(2)]PF(6) (LL = 1,10-phen (3a) and 4,4'-Me(2) 2,2'-bpy (3b)) are made from [(eta(6)-C(6)H(6))Os(micro-Cl)Cl](2) (1) either via transmetalation using the [Hg(phpy)(2)] organomercurial in MeOH or via the sp(2) C-H bond cleavage of 2-phenylpyridine (phpyH) in MeCN to afford [(eta(6) C(6)H(6))Os(phpy)Cl] or [(eta(6)-C(6)H(6))Os(phpy)(MeCN)]PF(6), respectively. The latter two react cleanly with LL to give 3a and 3b, the M(II/III) redox potentials of which equal 30 and -100 mV (vs Ag/AgCl), respectively. The electrochemically made Os(III) species oxidize rapidly reduced glucose oxidase. The second-order rate constant equals 1.1 x 10(7) M(-)(1) s(-)(1) for 3a at 25 degrees C, pH 7. PMID- 14552610 TI - Formation of a spiral-shaped inorganic-organic hybrid chain, [Cu(II)(2,2' bipy)(H(2)O)(2)Al(OH)(6)Mo(6)O(18)](n)()(n)()(-): influence of intra- and interchain supramolecular interactions. AB - A novel chainlike coordination polymer [Cu(II)(2,2' bipy)(H(2)O)(2)Al(OH)(6)Mo(6)O(18)](n)()(n)()(-), formed from a heteropolyanion [Al(OH)(6)Mo(6)O(18)](3)(-) as a building unit and a copper(II) complex fragment, [Cu(II)(2,2'-bipy)(H(2)O)(2)](2+), as a linker, provides the first example of an extended structure based on an Anderson type of polyanion and a transition metal complex with organic ligand. The intra- and interchain O-H.O hydrogen-bonding interactions are seemingly responsible for the spiral shape of this chain. Crystal data: triclinic space group Ponemacr;, a = 11.2253(18) A, b = 14.5194(17) A, c = 15.2672(10) A, alpha = 112.191(8) degrees, beta = 106.693(9) degrees, gamma = 93.916(13) degrees, and Z = 2. PMID- 14552611 TI - Octadecanuclear cluster or 1D polymer with [[ML](2)Nb(CN)(8)](n) motifs as a function of [ML] (M = Ni(II), n = 6; M = Mn(II), n = infinity; L = macrocycle). AB - A nanosized octadecaheteronuclear aggregate, [[NiL(2)](12)[Nb(CN)(8)](6)(H(2)O)(6)], and a 1-D coordination polymer, [[MnL(1)](2)[Nb(CN)(8)](H(2)O)]( infinity ), have been obtained by self-assembly between the octacyanometalate [Nb(CN)(8)](4)(-) and [ML](2+) complexes. The dimensionality of the supramolecular architectures was found to be controlled by the [ML] module for which the equatorial coordination sites are blocked by a macrocyclic ligand. The crystal structures and magnetic properties for both the compounds are described. PMID- 14552613 TI - Novel Ru-dioxolene complexes as potential electrochromic materials and NIR dyes. AB - A series of Ru(bpy)(2)-dioxolene complexes 1-4 (bpy = 2,2'- bipyridine) and corresponding Ru(dcb)(2)-dioxolene complexes 5-8 (dcbH(2) = 2,2'-bipyridine-4,4' dicarboxylic acid) have been prepared, and their spectroelectrochemical behavior in solution has been investigated. The complexes show reversible electrochemical behavior accompanied by a strong NIR absorption in their semiquinone forms due to a Ru(dpi) --> sq(pi) MLCT band. Complete quenching of the NIR absorption occurs both upon oxidation (to the quinone form) and upon reduction (to the catechol form) very close to 0 V. The color of the systems can be tuned by using a wide range of ligands. The complexes 5-8 can be anchored onto nanocrystalline inorganic semiconductors allowing incorporation into potential electrochromic devices. As a proof of principle, compound 8 has been adsorbed on nanocrystalline Sb-doped SnO(2) supported on FTO glass, and it displays reversibly switchable electrochromic behavior in the NIR. PMID- 14552612 TI - Cupric decamolybdodivanadophosphate. A coordination polymer heterogeneous catalyst for rapid, high conversion, high selectivity sulfoxidation using the ambient environment. AB - A new type of coordination network polymer involving the redox-active polyanion, PV(2)Mo(10)O(40)(5)(-), and bridging -Cu(II)(OH(2))(4)- units, [[(Cu(II)(OH(2))(4))(3)(OH)]PV(2)Mo(10)O(40)](n)() (1), has been characterized by X-ray crystallography and several other methods. It is the first efficient heterogeneous (insoluble) catalyst for selective and rapid sulfoxidation using only the ambient environment (air at room temperature). Catalytic activity is enhanced by soluble nitrate in nontoxic perfluoropolyether (PFPE) media. PMID- 14552614 TI - Excited-state energy-transfer dynamics of self-assembled imine-linked porphyrin dyads. AB - Toward the development of new strategies for the synthesis of multiporphyrin arrays, we have prepared and characterized (electrochemistry and static/time resolved optical spectroscopy) a series of dyads composed of a zinc porphyrin and a free base porphyrin joined via imine-based linkers. One dyad contains two zinc porphyrins. Imine formation occurs under gentle conditions without alteration of the porphyrin metalation state. Five imine linkers were investigated by combination of formyl, benzaldehyde, and salicylaldehyde groups with aniline and benzoic hydrazide groups. The imine-linked dyads are quite stable to routine handling. The excited-state energy-transfer rate from zinc to free base porphyrin ranges from (70 ps)(-)(1) to (13 ps)(-)(1) in toluene at room temperature depending on the linker employed. The energy-transfer yield is generally very high (>97%), with low yields of deleterious hole/electron transfer. Collectively, this work provides the foundation for the design of multiporphyrin arrays that self-assemble via stable imine linkages, have predictable electronic properties, and have comparable or even enhanced energy-transfer characteristics relative to those of other types of covalently linked systems. PMID- 14552615 TI - Excited-state energy-transfer dynamics in self-assembled triads composed of two porphyrins and an intervening Bis(dipyrrinato)metal complex. AB - The synthesis and characterization of various triads composed of a linear array of two zinc porphyrins joined via an intervening bis(dipyrrinato)metal(II) complex are reported. The preparation exploits the facile complexation of dipyrrins with divalent metal ions to give bis(dipyrrinato)metal(II) complexes [abbreviated (dp)(2)M]. Copper(II) and palladium(II) chelates of dipyrrins (available by oxidation of dipyrromethanes) were prepared in 50-80% yield. A one flask synthesis of bis(dipyrrinato)zinc(II) complexes was developed by oxidation of a dipyrromethane with DDQ or p-chloranil in the presence of Zn(OAc)(2).2H(2)O in THF ( approximately 80% yield). Three routes were developed for preparing porphyrin-dipyrrins: (1). Suzuki coupling of a boronate-substituted zinc porphyrin (ZnP) and bis[5-(4-iodophenyl)dipyrrinato]Pd(II) to give the (ZnP dp)(2)Pd triad (50% yield), followed by selective demetalation of the (dp)(2)Pd unit by treatment with 1,4-dithiothreitol under neutral conditions (71% yield); (2). oxidation of a porphyrin-dipyrromethane with p-chloranil in the presence of Zn(OAc)(2).2H(2)O followed by chromatography on silica gel (71% yield); and (3). condensation of a dipyrrin-dipyrromethane and a dipyrromethane-dicarbinol under InCl(3) catalysis followed by oxidation with DDQ (10-16% yield). Four triads of form (ZnP-dp)(2)Zn were prepared in 83-97% yield by treatment of a porphyrin dipyrrin with Zn(OAc)(2).2H(2)O at room temperature. Free base dipyrrins typically absorb at 430-440 nm, while the bis(dipyrrinato)metal complexes absorb at 460-490 nm. The fluorescence spectra/yields and excited-state lifetimes of the (ZnP-dp)(2)Zn triad in toluene show (1). efficient energy transfer from the bis(dipyrrinato)zinc(II) chromophore to the zinc porphyrins (98.5% yield), and (2). little or no quenching of the resulting excited zinc porphyrin relative to the isolated chromophore. Taken together, these results indicate that bis(dipyrrinato)zinc(II) complexes can serve as self-assembling linkers that further function as secondary light-collection elements in porphyrin-based light harvesting arrays. PMID- 14552616 TI - Cu(I) complexes of 3,3'-polymethylene bridged derivatives of 2,2'-bi-1,10 phenanthroline. AB - A series of 3,3'-polymethylene-2,2'-bi-1,10-phenanthrolines coordinate with Cu(I) to form dinuclear complexes [(CuL)(2)](2+). As the 3,3'-bridge is lengthened from two to four carbons, the ligand becomes more twisted about the 2,2'-bond, favoring dinuclear coordination. The distance between the two copper atoms varies from 2.92 A for the dimethylene-bridged system to 3.59 A for the tetramethylene bridge. Favorable pi-stacking interactions occur between opposing ligands and promote complex formation. Competition experiments indicate that self-recognition is important and only homoleptic complexes are observed. Under equilibrium conditions, formation of the tetramethylene-bridged complex appears to be the most favored while the dimethylene-bridged system is least favored. The intensity of the long wavelength metal-ligand charge-transfer absorption band decreases as the 3,3'-bridge is shortened. Interaction between the two copper centers is evidenced by a splitting of the oxidation wave, and this splitting increases as the Cu-Cu distance is decreased. PMID- 14552617 TI - Synthesis and comprehensive characterizations of new cis-RuL(2)X(2) (X = Cl, CN, and NCS) sensitizers for nanocrystalline TiO(2) solar cell using Bis-phosphonated bipyridine ligands (L). AB - The preparation and the properties of several ruthenium complexes of the general formula cis-RuL(2)X(2) with L = 2,2'-bipyridine-4,4'-bisphosphonic acid, L' = 2,2'-bipyridine-5,5'-bisphosphonic acid, and X = Cl, CN, or NCS are reported. The synthesis of these complexes relies on the preparation of the key intermediates cis-Ru(bipyridinebis(diethyl ester phosphonate))Cl(2). The ground-state second pK(a) values of the thiocyanato complexes were determined and are 6.0 and 6.1 for cis-RuL(2)(NCS)(2) and for cis-RuL'(2)(NCS)(2), respectively. For these species, (13)C NMR and IR demonstrate that the thiocyanato ligands are bound to Ru via the N atom. The new complexes exhibit a blue-shifted electronic absorption spectrum with respect to the analogous complexes containing carboxylic acid groups. Density functional theory molecular orbital calculations show that the LUMO of the bipyridine phosphonated ligands is at higher energy than the corresponding dicarboxylate complexes and that the thiocyanato ligands are not simple spectator ligands, whose role is to enrich electron density on the ruthenium, but are also involved in transitions from PiRu-NCS to Pibpy that extend the absorbance of the dye in the low energy part of the absorption spectrum. The photoaction spectra recorded in a sandwich regenerative photovoltaic cell indicate that the cyano and thiocyanato complexes containing the bipyridine substituted in 4,4' positions exhibit a 90-95% photoconversion efficiency on the MLCT band, whereas those containing the bipyridine substituted in 5,5' positions display lower efficiency (60-65%). The most efficient complex in the series is cis-RuL(2)(NCS)(2); however, its overall efficiency is about 30% lower than the analogue cis Ru(H(2)dcb)(2)(NCS)(2) (H(2)dcb = 2,2'-bipyridine-4,4'-dicarboxylic acid) due to a lower absorbance in the red part of the visible spectrum. PMID- 14552618 TI - New metal-rich sulfides Ni(6)SnS(2) and Ni(9)Sn(2)S(2) with a 2D metal framework: synthesis, crystal structure, and bonding. AB - Two new, metal-rich nickel-tin sulfides Ni(6)SnS(2) and Ni(9)Sn(2)S(2) were found by establishing phase relations in the ternary Ni-Sn-S system at 540 degrees C. Their single crystals were prepared by means of chemical vapor transport reactions. Single crystal X-ray diffraction was used for the determination of their crystal structures. Both compounds crystallize in a tetragonal system (I4/mmm, No. 139, Z = 2, a = 3.646(1) A, c = 18.151(8) A for Ni(6)SnS(2), and a = 3.678(1) A, c = 25.527(8) A for Ni(9)Sn(2)S(2)). Their crystal structures represent a new structure type and can be considered as assembled from bimetallic nickel-tin and nickel-sulfide slabs alternating along the crystallographic c axis. DFT band structure calculations showed the bonding within the bimetallic slabs to have a delocalized, multicenter nature, typical for metallic systems, and predominantly classical, pairwise bonding between nickel and sulfur. PMID- 14552619 TI - Nine hexagonal ca(5)pb(3)z phases in stuffed mn(5)si(3)-type structures with transition metal interstitial atoms z. Problems with classical valence States in possible zintl phases. AB - Ternary hexagonal Ae(5)Tt(3)Z phases have been obtained from high-temperature reactions (1000-1300 degrees C in Ta) only for Ae (alkaline-earth metal) = Ca, Tt (tetrel) = Pb, and Z = V, Cr, Mn, Fe, Co, Ni, Zn, Ru, or Cd. The hexagonal crystal structures (stuffed Mn(5)Si(3)-type, P6(3)/mcm, Z = 2) were refined for Z = Mn and Fe (a = 9.3580(3), 9.3554(5) A, c = 7.009(1), 7.009(1) A, respectively). In contrast, Ca(5)Pb(3)Z for Z = Cu or Ag form only with a trigonal structure (P3c1, Z = 2, a = 9.4130(3) A, c = 7.052(1) A for Cu) in which regular displacements of only the linear strings of Ca1 atoms occur. The existence of these compounds stands in contrast to the nonexistence of all binary Ae(5)Tt(3) products from Ca to Ba (Ae) and Si to Pb (Tt) with a Mn(5)Si(3)-type structure. Therefore, it once seemed attractive to consider the Z elements in these Ca(5)Pb(3)Z compounds as reducing agents (electron donors). The Mn and Fe structures appropriately exhibit greatly enlarged antiprismatic calcium cavities about Z. Other indications of relatively electron-poor environments around Fe are found in its properties, which include soft ferromagnetism with an elevated magnetic moment (6.3 micro(B)) and a large Fe 3p(3/2) binding energy relative to that in La(5)Ge(3)Fe, La(15)Ge(9)Fe, etc. The Ca(5)Pb(3)Mn phase exhibits metallic behavior (rho(295) = 135 microOmega cm) and temperature-independent Pauli paramagnetism. These properties are supported by ab initio band structure calculations for Ca(5)Pb(3)Mn, which show strong Ca-Pb bonding and a broad Pb based band, with appreciable Ca-Mn and Ca-Pb bonding states at and above E(F). Distortion of the Cu analogue gives strengthened Ca-Pb bonding and reduced Cu-Ca1 repulsions. A Zintl phase description of these compounds and some related compounds in terms of closed Pb bands is not appropriate. PMID- 14552620 TI - Tris(bis(trimethylsilyl)amido)samarium: X-ray structure and DFT study. AB - The compound Sm[N(SiMe(3))(2)](3) has been investigated experimentally by X-ray crystallography and computationally by DFT methods. The structure is analogous to that of other tris[bis(trimethylsilyl)amido]lanthanides, featuring positional disorder of the metal atom above and below the plane defined by the three N donor atoms, resulting in a trigonal pyramidal configuration. One of the methyl groups of each amido ligand is placed above the apex of the pyramid at close distance to the metal center suggesting the presence of agostic interactions. The DFT calculations have been carried out on the real molecule and on a Si[N(SiH(3))(SiH(2)Me)](3) model where the unique Me group was placed above the apex of the pyramid to probe the agostic interaction. In both cases, the optimized geometry reproduces very well the experimental structure and indicates the presence of beta-Si-C agostic interactions. A comparison of the optimized geometries obtained in the presence/absence of the Sm d and the Si d orbitals serves to illustrate the relevance of these orbitals for (i). the establishment of the pyramidal configuration at Sm, (ii). the Sm-N bond length, and (iii). the Sm-(beta-Si-C) bond length. The bonding analysis, which was carried out by both Mulliken and NBO methods, not only confirms the importance of the metal d orbitals for the Sm-N and Sm-(beta-Si-C) chemical bonding but also illustrates the relevance of electrostatic terms in the agostic interaction. Sm-N and N-Si pi bonding is present according to the bonding analysis but is not important for enforcing the planar configuration at N, nor the pyramidal configuration at Sm. PMID- 14552621 TI - Pi bonding and negative hyperconjugation in mono-, di-, and triaminoborane, alane, -gallane, and -indane. AB - A systematic quantum chemical investigation of mono-, di-, and triaminoborane, alane, -gallane, and -indane is carried out to determine quantitatively the effects of pi bonding and negative hyperconjugation on structures, energetics, and rotational barriers in these systems. Pi bonding plays a significant role in the aminoborane compounds, but becomes rapidly less significant in the aminoalanes, -gallanes, and -indanes. For each main-group metal X investigated, X N rotational barriers are found to be essentially equal depending only on the number of remaining in-plane amino groups. The contribution of negative hyperconjugation to reducing rotational barriers, as assessed from natural bond orbital (NBO) delocalization energies, is independent of the pyramidalization of the out-of-plane amino group, and is also dependent only on the number of rotated groups. Optimized tris[bis(trimethylsilyl)amino]-substituted structures of boron, aluminum, gallium, and indium are found to compare quite well with available experimental structural data, and exhibit X-N torsion angles that are independent of the central metal atom. PMID- 14552622 TI - Density functional theory study of nine-atom germanium clusters: effect of electron count on cluster geometry. AB - Density functional theory (DFT) at the hybrid B3LYP level has been applied to the germanium clusters Ge(9)(z) clusters (z = -6, -4, -3, -2, 0, +2, and +4) starting from three different initial configurations. Double-zeta quality LANL2DZ basis functions extended by adding one set of polarization (d) and one set of diffuse (p) functions were used. The global minimum for Ge(9)(2)(-) is the tricapped trigonal prism expected by Wade's rules for a 2n + 2 skeletal electron structure. An elongated tricapped trigonal prism is the global minimum for Ge(9)(4)(-) similar to the experimentally found structure for the isoelectronic Bi(9)(5+). However, the capped square antiprism predicted by Wade's rules for a 2n + 4 skeletal electron structure is only 0.21 kcal/mol above this global minimum indicating that these two nine-vertex polyhedra have very similar energies in this system. Tricapped trigonal prismatic structures are found for both singlet and triplet Ge(9)(6)(-), with the latter being lower in energy by 3.66 kcal/mol and far less distorted. The global minimum for the hypoelectronic Ge(9) is a bicapped pentagonal bipyramid. However, a second structure for Ge(9) only 4.54 kcal/mol above this global minimum is the C(2)(v)() flattened tricapped trigonal prism structure found experimentally for the isoelectronic Tl(9)(9)(-). For the even more hypoelectronic Ge(9)(2+), the lowest energy structure consists of an octahedron fused to two trigonal bipyramids. For Ge(9)(4+), the global minimum is an oblate (squashed) pentagonal bipyramid with two pendant Ge vertices. PMID- 14552623 TI - Reversible guest exchange and ferrimagnetism (T(C) = 60.5 K) in a porous cobalt(II)-hydroxide layer structure pillared with trans-1,4 cyclohexanedicarboxylate. AB - The synthesis, characterization, and reversible guest-exchange chemistry of a new porous magnetic material that orders ferrimagnetically at 60.5 K are described. The material, Co(5)(OH)(8)(chdc).4H(2)O (chdc = trans-1,4 cyclohexanedicarboxylate), contains tetrahedral-octahedral-tetrahedral Co(II) hydroxide layers of composition Co((oct))(3)Co((tet))(2)(OH)(8) that are linked together by bis(unidentate) chdc pillars. Noncoordinated water molecules occupy 1 D channels situated between the chdc pillars. The material remains monocrystalline during dehydration from Co(5)(OH)(8)(chdc).4H(2)O (CDCC.4H(2)O) to Co(5)(OH)(8)(chdc) (CDCC) via an intermediate Co(5)(OH)(8)(chdc).2H(2)O (CDCC.2H(2)O) upon heating or evacuation. In-situ single crystal and powder X-ray diffraction analyses indicate that the interlayer spacing decreases in two steps, each corresponding to the loss of two water molecules per formula unit as determined by thermogravimetry. The single crystal structure of the fully dehydrated material, CDCC, has no void volume due to a tilting of the pillars and 9% decrease of the interlayer spacing with water removal. Exposure of CDCC to air causes rapid rehydration of this material to CDCC.4H(2)O, as determined by single crystal X-ray diffraction, powder X-ray diffraction, thermogravimetry, and vibrational spectroscopy. Both the hydrated and dehydrated forms order magnetically below 60.5 K. The susceptibility data are consistent with ferrimagnetic behavior, and the value of the saturation magnetization at 2 K (ca. 2 micro(B)) is explained by a model of two sublattices, one comprising three octahedral cobalt atoms and another comprising two tetrahedral cobalt atoms. There is an enhanced 2-D correlation within the layer at temperatures just above the Curie temperature, as seen by nonlinearity in the ac susceptibility data and remanence in the isothermal magnetization. The crossover from 2-D to 3-D ordering occurs at T(C). The large anisotropy in the magnetization data on a single crystal suggests either a 2-D Ising or an XY magnet while the critical exponent of 0.25 is in favor of the latter. Both magnetization data in a small field in the ac and dc mode and isothermal magnetization data provide evidence of a further change in behavior at 23 K, which may originate from a reorientation of the moments within the layer. Variation of the pillar and of the guest-exchange chemistry, including the exchange of magnetic guests such as O(2), offers the possibility of tailoring the magnetic properties of this material. PMID- 14552624 TI - Silicon-silica nanowires, nanotubes, and biaxial nanowires: inside, outside, and side-by-side growth of silicon versus silica on zeolite. AB - It was demonstrated that zeolite can be used as a pseudo-template to grow very fine and uniform silicon nanostructures via disproportionation reaction of SiO by thermal evaporation. Three distinct types of composite nanowires and nanotubes of silicon and silica were grown on the surfaces of zeolite Y pellets. The first type is formed by an ultrafine crystalline silicon nanowire sheathed by an amorphous silica tube (a silicon nanowire inside a silica nanotube). The second type is formed by a crystalline silicon nanotube filled with amorphous silica (a silicon nanotube outside a silica nanowire). The third type is a biaxial silicon silica nanowire structure with side-by-side growth of crystalline silicon and amorphous silica. These silicon nanostructures exhibit unusually intense photoluminescence (in comparison to ordinary silicon nanowires). PMID- 14552625 TI - Solid state coordination chemistry: one-, two-, and three-dimensional materials constructed from molybdophosphonate subunits linked through binuclear copper tetra-2-pyridylpyrazine groups. AB - The hydrothermal reactions of MoO(3), an appropriate Cu(II) source, tetra-2 pyridylpyrazine (tpypyz), and phosphoric acid and/or an organophosphonate yielded a series of organic-inorganic hybrid materials of the copper-molybdophosphonate family. A common feature of the structures is the entrainment within the extended architectures of chemically robust [Mo(5)O(15)(O(3)PR)(2)](4)(-) clusters as molecular building blocks. The cluster is a characteristic feature of the one dimensional materials [[Cu(2)(tpypyz)(H(2)O)(3)]Mo(5)O(15)(HPO(4))(O(3)PCH(2)CO(2)H)].H(2)O (1.H(2)O) and [[Cu(2)(tpypyz)(H(2)O)]Mo(5)O(15)(O(3)PC(6)H(5))(2)].2H(2)O (2.2H(2)O), the two-dimensional network [[Cu(2)(tpypyz)(H(2)O)(3)]Mo(5)O(15)(HPO(4))(2)].2H(2)O (5.2H(2)O) and the three-dimensional frameworks [[Cu(2)(tpypyz)(H(2)O)(2)]Mo(5)O(15)[O(3)P(CH(2))(n)()PO(3)]].xH(2)O [n = 3, x = 2.25 (6.2.25H(2)O); n = 4, x = 0.33 (7.0.33H(2)O)]. In the case of methylenediphosphonate as the phosphorus component, the unique chelating nature of the ligand precludes formation of the pentamolybdate core, resulting in the chain structures [[Cu(2)(tpypyz)(H(2)O)]Mo(3)O(8) (HO(3)PCH(2)PO(3))(2)].8H(2)O (3.8H(2)O) and [[Cu(2)(tpypyz)(H(2)O)](2)(Mo(3)O(8))(2)(O(3)PCH(2)PO(3))(3)].16.9H(2)O (4.16.9H(2)O). For structures 1-7, the secondary metal-ligand building block is the binuclear [Cu(2)(tpypyz)(H(2)O)(x)](4+) cluster. There is considerable structural versatility as a result of the variability in the number of attachment sites at the phosphomolybdate clusters, the coordination geometry of the Cu(II), which may be four-, five-, or six-coordinate, the extent of aqua ligation, and the participation of phosphate oxygen atoms as well as molybdate oxo groups in bonding to the copper sites. Crystal data: 1.H(2)O, C(26)H(28)N(6)Cu(2)Mo(5)O(28)P(2), monoclinic C2/c, a = 42.497(2) A, b = 10.7421(4) A, c = 20.5617(8) A, beta = 117.178(1) degrees, V = 8350.1(5) A(3), Z = 8; 2.2H(2)O, C(36)H(32)N(6)Cu(2)Mo(5)O(24)P(2), monoclinic P2(1)/c, a = 11.2478(7) A, b = 19.513(1) A, c = 21.063(1) A, beta = 93.608(1) degrees, V = 4613.7(5) A(3), Z = 4; 3.8H(2)O, C(26)H(40)N(6)Cu(2)Mo(3)O(29)P(4), monoclinic C2/c, a = 32.580(2) A, b = 17.8676(9) A, c = 15.9612(8) A, beta = 104.430(1) degrees, V = 8993.3(8) A(3), Z = 8; 4.16.9H(2)O, C(51)H(71.75)Cu(4)Mo(6)N(12)O(51)P(6), monoclinic P2(1)/c, a = 27.929(3) A, b = 12.892(2) A, c = 22.763(3) A, beta = 90.367(2) degrees, V = 8195.7(2) A(3), Z = 4;( )()5.2H(2)O, C(24)H(28)N(6)Cu(2)Mo(5)O(28)P(2), monoclinic P2(1)/n, a = 11.3222(4) A, b = 18.7673(7) A, c = 19.4124(7) A, beta = 98.819(1) degrees, V = 4076.1(3) A(3), Z = 4; 6.2.25H(2)O, C(27)H(28.5)N(6)Cu(2)Mo(5)O(24.25)P(2), monoclinic C2/c, a = 12.8366(5) A, b = 18.4221(8) A, c = 34.326(1) A, beta = 100.546(1) degrees, V = 7980.1(6) A(3), Z = 8; 7.(1)/(3)H(2)O, C(28)H(28.7)N(6)Cu(2)Mo(5)O(23.3)P(2), monoclinic C2/c, a = 12.577(1) A, b = 18.336(1) A, c = 36.476(3) A, beta = 91.929(2) degrees, V = 8407.3 A(3), Z = 8. PMID- 14552626 TI - Orange luminescence and structural properties of three isostructural halocyclohexylisonitrilegold(I) complexes. AB - The preparation of three isonitrile complexes (CyNC)Au(I)Cl, (CyNC)Au(I)Br, and (CyNC)Au(I)I, along with their structural and spectral characterization, are reported. X-ray crystal structures reveal that these crystallize in the same space group and have closely related structures. The structures involve pleated chains of linear, two-coordinate monomers that are arranged in a head-tail fashion. However, these chains vary significantly in the degree of aurophilic interactions among the individual molecules. Thus, (CyNC)Au(I)Cl forms infinite chains with alternating Au...Au distances of 3.3894(7) and 3.5816(7) A. Within the chains of (CyNC)Au(I)Br, however, the alternation of Au.Au distances is more pronounced so that there are dimers, with an Au.Au distance of 3.4864(9) A, and neighboring gold centers at 3.7036(9) A. In (CyNC)Au(I)I, the gold-gold contacts do not lie within the range of significant aurophilic bonding. The closest Au...Au distance is 3.7182(11) A while every other Au...Au distance is 3.9304(12) A. The steric factor of the X ligand and dipole-dipole interactions between the antiparallel complexes is much more significant than aurophilic interactions in governing the self-association of the complexes in this series. The colorless crystals of each solid display an orange luminescence band with a strikingly large Stokes' shift ( approximately 21000 cm(-)(1), 2.6 eV). However, considerable care had to be taken to ensure that the crystals used for the study of the luminescence were free of a surface impurity that produced a turquoise green luminescence in (CyNC)Au(I)Cl. The diffuse reflectance spectra for the solids show a similar three-band pattern in the 200-330 nm range. PMID- 14552627 TI - Nickel(II) cyclidenes with appended ethylpyridine receptor centers as molecular tweezers for dicarboxylic acids. AB - A series of 14-, 15-, and 16-membered nickel(II) cyclidene macrocycles appended with 2-aminoethyl(2-pyridine) receptors I-III, respectively, were prepared and characterized by X-ray crystallography and NMR techniques. The 14- and 15 membered macrocycles I and II exist in a planar or extended Z-configuration, whereas the 16-membered macrocycle III was saddle shaped and had two asymmetric configurations in the unit cell (IIIa in a "capped" configuration and IIIb in an "open" configuration). Variable-temperature (1)H NMR studies of III in CD(3)CN were conducted (25-65 degrees C), and at room temperature, the interconversion between capping and uncapping is slow on the NMR time scale, resulting in a broad spectrum, whereas at 65 degrees C, interconversion was fast. (1)H NMR binding studies indicated I-III bind unsaturated dicarboxylic acids in a 1:1 stoichiometry with binding constants approaching 400 M(-)(1) in CD(3)CN, and the binding strength was dependent on the shape of the macrocyclic cyclidene platforms, whereas monocarboxylic acids were not bound. Generally, the planar 14 membered cyclidene I bound diacids the weakest and the 16-membered cyclidene III bound diacids the strongest. The presence of nuclear Overhauser effect spectrometry cross peaks in a 20 mM solution of 1:1 II-maleic acid indicates that the binding mode is ditopic with the guest being encapsulated by the aminoethylpyridine arms above the macrocyclic framework. PMID- 14552628 TI - DFT study on the reed diethylaluminum cation-like system: structure and bonding in Et(2)Al(CB(11)H(6)X(6)) (X = Cl, Br). AB - Electronic and molecular structure has been investigated in the diethylaluminum cation-like system Et(2)Al(CB(11)H(6)X(6)) (1, X = Cl; 2, X = Br) and neutral compounds AlX(3) (X = Cl, Br, Me, C(6)H(5)) with DFT B3LYP and BP86 levels of theory. The calculated geometries of Et(2)Al(CB(11)H(6)X(6)) (1, X = Cl; 2, X = Br) are in excellent agreement with those determined experimentally by X-ray crystallography. The Al-X bond distances 2.442, 2.445 A in 1 and 2.579, 2.589 A in 2 are longer than those expected for single bonds based on covalent radius predictions (Al-Cl = 2.15 A and Al-Br = 2.32 A) and those observed for bridged Al X-Al bonds (2.21 A in Al(2)Cl(6), 2.33 A in Al(2)Br(6)) and are close to sum of ionic radii of Al(3+) and X(-) (Al-Cl = 2.35 A and Al-Br = 2.50 A). The optimized geometries of the neutral compounds AlX(3) (X = Cl, Br, Me(3), C(6)H(5)) at BP86/TZ2P show Al-Cl = 2.088 A in AlCl(3), Al-Br = 2.234 A in AlBr(3), Al-C = 1.973 A in AlMe(3), Al-C = 2.255 A in Al(C(6)F(5))(3). These bond distances are similar to those expected for single bonds based on covalent radius predictions. The calculated charge distribution indicates that the aluminum atom carries a significant positive charge while the ethyl and carborane groups are negatively charged. The Cl and Br atoms in compounds 1 and 2 are slightly positive while, in neutral compounds AlX(3) (X = Cl, Br, Me(3), C(6)H(5)), X is negatively charged. Energy decomposition analysis of Et(2)Al(delta)(+)(carborane)(delta)(-) shows that the bonding between the fragments is more than half electrostatic. The ionic character of the Al...Cl bonds in compound 1(59.8%) is greater than the Al.Br bonds in the compound 2 (57.9%). This quantifies and gives legitimacy to the designation of these types of compounds as "ion-like". The Al-X bonding in AlX(3) is mainly covalent with percentage ionic character 28.2% in AlCl(3), 31.5% in AlBr(3), 25.6% in AlMe(3), 18.4% in Al(C(6)F(5))(3). PMID- 14552629 TI - Metal complexes with a new N(4)O(3) amine pendant-armed macrocyclic ligand: synthesis, characterization, crystal structures, and fluorescence studies. AB - The synthesis of a new oxaaza macrocyclic ligand, L, derived from O(1),O(7)-bis(2 formylphenyl)-1,4,7-trioxaheptane and tren containing an amine terminal pendant arm, and its metal complexation with alkaline earth (M = Ca(2+), Sr(2+), Ba(2+)), transition (M = Co(2+), Ni(2+), Cu(2+), Zn(2+), Cd(2+)), post-transition (M = Pb(2+)), and Y(3+) and lanthanide (M = La(3+), Er(3+)) metal ions are reported. Crystal structures of [H(2)L](ClO(4))(2).3H(2)O, [PbL](ClO(4))(2), and [ZnLCl](ClO(4)).H(2)O are also reported. In the [PbL] complex, the metal ion is located inside the macrocyclic cavity coordinated by all N(4)O(3) donor atoms while, in the [ZnLCl] complex, the metal ion is encapsulated only by the nitrogen atoms present in the ligand. pi-pi interactions in the [H(2)L](ClO(4))(2).3H(2)O and [PbL](ClO(4))(2) structures are observed. Protonation and Zn(2+), Cd(2+), and Cu(2+) complexation were studied by means of potentiometric, UV-vis, and fluorescent emission measurements. The 10-fold fluorescence emission increase observed in the pH range 7-9 in the presence of Zn(2+) leads to L as a good sensor for this biological metal in water solution. PMID- 14552630 TI - Synthesis of a copper [3]rotaxane able to function as an electrochemically driven oscillatory machine in solution, and to form SAMs on a metal surface. AB - Two new copper [3]rotaxanes have been synthesized. The axes are identical for both compounds and incorporate two bidentate chelates joined together by a disulfide bridge. The rings contain either the single phen (phen = 1,10 phenanthroline) chelate or two different chelates (phen and terpy; terpy = 2,2',6',2' '-terpyridine, a tridentate chelate). The key intermediates for both synthetic routes are semi-rotaxanes obtained in high yields using the three dimensional effect of copper(I). In the case where the wheels are heterobischelating macrocycles, large molecular motions, namely rotation or oscillation of the wheels around the axle, have been induced electrochemically. Anchoring of these two copper [3]rotaxanes on a gold electrode was carried out by standard procedures. Cleavage of the disulfide bridge and formation of monolayers of rotaxanes were evidenced by cyclic voltammetry. The adsorbed rotaxanes can be viewed as copper [2]rotaxanes for which the gold electrode surface acts as a stopper linked to one end of their axes. PMID- 14552631 TI - Speciation and coordination chemistry of uranyl(VI)-citrate complexes in aqueous solution. AB - The pH dependence of uranyl(VI) complexation by citric acid was investigated using Raman and attenuated total reflection FTIR spectroscopies and electrospray ionization mass spectrometry. pH-dependent changes in the nu(s)(UO(2)) envelope indicate that three major UO(2)(2+)-citrate complexes with progressively increasing U=O bond lengths are present over a range of pH from 2.0 to 9.5. The first species, which is the predominant form of uranyl(VI) from pH 3.0 to 5.0, contains two UO(2)(2+) groups in spectroscopically equivalent coordination environments and corresponds to the [(UO(2))(2)Cit(2)](2)(-) complex known to exist in this pH range. At pH values >6.5, [(UO(2))(2)Cit(2)](2)(-) undergoes an interconversion to form [(UO(2))(3)Cit(3)](3)(-) and (UO(2))(3)Cit(2). ESI-MS studies on solutions of varying uranyl(VI)/citrate ratios, pH, and solution counteranion were successfully used to confirm complex stoichiometries. Uranyl and citrate concentrations investigated ranged from 0.50 to 50 mM. PMID- 14552632 TI - Synthesis and characterization of a new asymmetric bis-porphyrinato lanthanide complex presenting mixed hydrophilic-hydrophobic properties and its precursor form. AB - The synthesis and spectroscopic characterization of a new family of heteroleptic porphyrinate double-deckers are reported. The investigated compounds are represented by the formulae La(III)H(TPyP)(TPP) and [La(III)(TMePyP)(TPP)]I(3). UV-vis spectroscopy of the title complexes confirms the presence of a strong pi pi interaction between the macrocycles in each derivative. With (1)H and 2-D NMR data, we were able to distinguish two major NMR regions: the endo, between the bonded macrocycles, and the exo, outside the macrocycles, which are characteristic features of porphyrinic double-deckers. Finally, the electrochemical study confirms the strong pi-pi interaction for La(III)H(TPyP)(TPP) and completes this first approach for the investigation of this new family of derivatives. PMID- 14552633 TI - New beta cis folded organocobalt derivatives with a salen-type ligand. AB - The reduction of [Co(III)(tmsalen)py(2)](+)ClO(4)(-), where tmsalen = 4,4',7,7' tetramethylsalen, with NaBH(4)/PdCl(2) in alkaline methanolic solution, followed by the oxidative addition of CH(2)ClI, leads to the expected trans organometallic dimeric species 1, [CH(2)ClCo(tmsalen)](2), provided that the product is recovered from the reaction mixture immediately after the completion of the reaction. If 1 is left for longer time in contact with the reaction mixture, the intramolecular reaction of the axial chloromethyl group with the equatorial chelate leads to the formation of the monocationic complex 2, containing a seven membered ring. In this complex the novel tetradentate ligand coordinates Co in a cis fashion, the other two positions being occupied by one py and one water molecule. The resulting complex is chiral, even if the reaction product is a racemic compound. The unidentate ligands of 2 have been exchanged quantitatively for N-MeIm, and the resulting complex 3 still maintains the beta cis geometry. Therefore, 2 may be considered the precursor of a new class of organocobalt derivatives with a folded tetradentate ligand and two adjacent exchangeable sites. On the basis of the geometry of the tetradentate Schiff bases in complexes, where they adopt a planar geometry, it was suggested that there is a significant electron density delocalization involving the metal center over the two chemically equivalent moieties of Co(chel). Comparison of the geometry of the planar salicylaldiminate (sal) moiety with that of the cyclized methoxy-iminate one (imi) in 2 and 3 strongly supports that the delocalization, still present in sal, is essentially either lost or strongly reduced in imi. PMID- 14552634 TI - Iron nitrosyls of a pentadentate ligand containing a single carboxamide group: syntheses, structures, electronic properties, and photolability of NO. AB - Three iron complexes of a pentadentate ligand N,N-bis(2-pyridylmethyl)amine-N ethyl-2-pyridine-2-carboxamide (PaPy(3)H, H is the dissociable amide proton) have been synthesized. All three species, namely, two nitrosyls [(PaPy(3))Fe(NO)](ClO(4))(2) (2) and [(PaPy(3))Fe(NO)](ClO(4)) (3) and one nitro complex [(PaPy(3))Fe(NO(2))](ClO(4)) (4), have been structurally characterized. These complexes provide the opportunity to compare the structural and spectral properties of a set of isostructural [Fe-NO](6,7) complexes (2 and 3, respectively) and an analogous genuine Fe(III) complex with an "innocent" sixth ligand ([(PaPy(3))Fe(NO(2))](ClO(4)), 4). The most striking difference in the structural features of 2 and 3 is the Fe-N-O angle (Fe-N-O = 173.1(2) degrees in the case of 2 and 141.29(15) degrees in the case of 3). The clean (1)H NMR spectrum of 2 in CD(3)CN reveals its S = 0 ground state and confirms its [Fe NO](6) configuration. The binding of NO at the non-heme iron center in 2 is completely reversible and the bound NO is photolabile. Mossbauer data, electron paramagnetic resonance signal at g approximately 2.00, and variable temperature magnetic susceptibility measurements indicate the S = (1)/(2) spin state of the [Fe-NO](7) complex 3. Analysis of the spectroscopic data suggests Fe(II)-NO(+) and Fe(II)-NO(*) formulations for 2 and 3, respectively. The bound NO in 3 does not show any photolability. However, in MeCN solution, it reacts rapidly with dioxygen to afford the nitro complex 4, which has also been synthesized independently from [(PaPy(3))Fe(MeCN)](2+) and NO(2)(-). Nucleophilic attack of hydroxide ion to the N atom of the NO ligand in 2 in MeCN in the dark gives rise to 4 in high yield. PMID- 14552635 TI - Investigation of the reduced high-potential iron-sulfur protein from chromatium vinosum and relevant model compounds: a unified picture of the electronic structure of [Fe(4)S(4)](2+) systems through magnetic and optical studies. AB - Magnetization measurements and variable temperature optical spectroscopy have been used to investigate, within the 4-300 K temperature range, the electronic structure of the reduced high-potential iron protein (HiPIP) from Chromatium vinosum and the model compounds (Cat)(2)[Fe(4)S(4)(SR)(4)], where RS(-) = 2,4,6 triisopropylphenylthiolate (1), 2,6-diphenylphenylthiolate (2), diphenylmethylthiolate (3), 2,4,6-triisopropylbenzylthiolate (4, 4'), 2,4,6 triphenylbenzylthiolate (5, 5'), 2,4,6-tri-tert-butylbenzylthiolate (6), and Cat(+) = (+)NEt(4) (1, 2, 3, 4', 5', 6), (+)PPh(4) (4, 5). The newly synthesized 2(2)(-), 3(2)(-), 5(2)(-), and 6(2)(-) complexes are, as 1(2)(-) and 4(2)(-), excellent models of the reduced HiPIPs: they exhibit the [Fe(4)S(4)](3+/2+) redox couple, because of the presence of bulky ligands which stabilize the [Fe(4)S(4)](3+) oxidized core. Moreover, the presence of SCH(2) groups in 4(2)( ), 5(2)(-), and 6(2)(-), as in the [Fe(4)S(4)] protein cores, makes them good biomimetic models of the HiPIPs. The X-ray structure of 2 is reported: it crystallizes in the orthorhombic space group Pcca with no imposed symmetry and a D(2)(d)()-distorted geometry of the [Fe(4)S(4)](2+) core. Fit of the magnetization data of the reduced HiPIP and of the 1, 2, 3, 4, 5, and 6 compounds within the exchange and double exchange theoretical framework leads to exchange coupling parameters J = 261-397 cm(-)(1). A firm determination of the double exchange parameters B or, equivalently, the transfer integrals beta = 5B could not be achieved that way. The obtained |B| values remain however high, attesting thus to the strength of the spin-dependent electronic delocalization which is responsible for lowest lying electronic states being characterized by delocalized mixed-valence pairs of maximum spin (9)/(2). Electronic properties of these systems are then accounted for by the population of a diamagnetic ground level and excited paramagnetic triplet and quintet levels, which are respectively J and 3J above the ground level. Optical studies of 1, 2, 4', 5', and 6 but also of (NEt(4))(2)[Fe(4)S(4)(SCH(2)C(6)H(5))(4)] and the isomorph (NEt(4))(2)[Fe(4)S(4)(S-t-Bu)(4)] and (NEt(4))(2)[Fe(4)Se(4)(S-t-Bu)(4)] compounds reveal two absorption bands in the near infrared region, at 705-760 nm and 1270-1430 nm, which appear to be characteristic of valence-delocalized and ferromagnetically coupled [Fe(2)X(2)](+) (X = S, Se) units. The |B| and |beta| values can be directly determined from the location at 10|B| of the low-energy band, and are respectively of 699-787 and 3497-3937 cm(-)(1). Both absorption bands are also present in the 77 K spectrum of the reduced HiPIP, at 700 and 1040 nm (Cerdonio, M.; Wang, R.-H.; Rawlings, J.; Gray, H. B. J. Am. Chem. Soc. 1974, 96, 6534-6535). The blue shift of the low-energy band is attributed to the inequivalent environments of the Fe sites in the protein, rather than to an increase of |beta| when going from the models to the HiPIP. The small differences observed in known geometries of [Fe(4)S(4)](2+) clusters, especially in the Fe-Fe distances, cannot probably lead to drastic changes in the direct Fe-Fe interactions (parameter beta) responsible for the delocalization phenomenon. These differences are however magnetostructurally significant as shown by the 261 397 cm(-)(1) range spanned by J. The cluster's geometry, hence the efficiency of the Femicro(3)-S-Fe superexchange pathways, is proposed to be controlled by the more or less tight fit of the cluster within the cavity provided by its environment. PMID- 14552637 TI - Hydrothermal synthesis in the system Ni(OH)(2)-NiSO(4): nuclear and magnetic structures and magnetic properties of Ni(3)(OH)(2)(SO(4))(2)(H(2)O)(2). AB - We present the synthesis, characterization by DT-TGA and IR, single crystal X-ray nuclear structure at 300 K, nuclear and magnetic structure from neutron powder diffraction on a deuterated sample at 1.4 K, and magnetic properties as a function of temperature and magnetic field of Ni(3)(OH)(2)(SO(4))(2)(H(2)O)(2). The structure is formed of chains, parallel to the c-axis, of edge-sharing Ni(1)O(6) octahedra, connected by the corners of Ni(2)O(6) octahedra to form corrugated sheets along the bc-plane. The sheets are connected to one another by the sulfate groups to form the 3D network. The magnetic properties measured by ac and dc magnetization, isothermal magnetization at 2 K, and heat capacity are characterized by a transition from a paramagnet (C = 3.954 emu K/mol and theta = 31 K) to a canted antiferromagnet at T(N) = 29 K with an estimated canting angle of 0.2-0.3 degrees. Deduced from powder neutron diffraction data, the magnetic structure is modeled by alternate pairs of Ni(1) within a chain having their moments pointing along [010] and [010], respectively. The moments of Ni(2) atoms are oppositely oriented with respect to their adjacent pairs. The resulting structure is that of a compensated arrangement of moments within one layer, comprising one ferromagnetic and three antiferromagnetic superexchange pathways between the nickel atoms. PMID- 14552636 TI - Fine-tuning the weak-link approach: effect of ligand electron density on the formation of rhodium(I) and iridium(I) metallomacrocycles. AB - A novel bis(phosphinoalkyl-thioether)arene ligand with a fluorinated aryl group (1,4-(Ph(2)PCH(2)CH(2)S)(2)C(6)F(4)) has been synthesized. This ligand has been used to prepare symmetric bimetallic structures with Rh(I) and Ir(I) metal centers in high yield. Unlike their nonfluorinated counterparts, these complexes can be opened into large macrocyclic structures through straightforward ligand (i.e., carbon monoxide, nitriles, and isocyanides) substitution reactions at the metal-thioether linkage. In addition, the symmetric bimetallic structures have been shown to react with appropriately sized bifunctional aromatic molecules to form three-tiered host-guest structures. PMID- 14552638 TI - Metal-centered ferrocene clusters from 5-ferrocenylpyrimidine and ferrocenylpyrazine. AB - A series of metal-centered ferrocene compounds has been designed by using 5 ferrocenylpyrimidine (L1) and ferrocenylpyrazine (L2). These ligands, when combined with transition metal salts, produce mixed-metal polynuclear complexes with structural diversity. Reaction of L1 with M(SCN)(2) (M = Ni, Co) produces the pinwheel-like 4:1 complexes (L1)(4).M(SCN)(2), while reactions of L1 and L2 with Cu(NO(3))(2) give the 3:1 complex (L1)(3).Cu(NO(3))(2) and the 2:1 complex (L2)(2).Cu(NO(3))(2), respectively. Reactions of L1 and L2 with M(hfac)(2) (hfac = 1,1,1,5,5,5-hexafluoroacetylacetonate, M = Mn, Ni, Cu, Zn) produce 2:1 complexes (L)(2).M(hfac)(2) with cis and trans configurations, respectively. Crystal structures as well as solid-state electrochemical properties of these redox active complexes were investigated. PMID- 14552639 TI - Chromium(II) and chromium(III) complexes supported by tris(2-pyridylmethyl)amine: synthesis, structures, and reactivity. AB - This report describes the synthesis, structural characterization, and polymerization behavior of a series of chromium(II) and chromium(III) complexes ligated by tris(2-pyridylmethyl)amine (TPA), including chromium(III) organometallic derivatives. For instance, the combination of TPA with CrCl(2) yields monomeric (TPA)CrCl(2) (1). A similar reaction of CrCl(2) with TPA, followed by chloride abstraction with NaBPh(4) or NaBAr(F)(4) (Ar(F) = 3,5 (CF(3))(2)C(6)H(3)), provides the weakly associated cationic dimers [(TPA)CrCl](2)[BPh(4)](2) (2A) and [(TPA)CrCl](2)[BAr(F)(4)](2) (2B), respectively. X-ray crystallographic analysis reveals that each chromium(II) center in 1, 2A, and 2B is a tetragonally elongated octahedron; such Jahn-Teller distortions are consistent with the observed high spin (S = 2) electronic configurations for these chromium(II) complexes. Likewise, reaction of CrCl(3)(THF)(3) with TPA, followed by anion metathesis with NaBPh(4) or NaBAr(F)(4), yields the monomeric, cationic chromium(III) complexes [(TPA)CrCl(2)][BPh(4)] (4A) and [(TPA)CrCl(2)][BAr(F)(4)] (4B), respectively. Treatment of 4A with methyl and phenyl Grignard reagents produces the cationic chromium(III) organometallic derivatives [(TPA)Cr(CH(3))(2)][BPh(4)] (5) and [(TPA)CrPh(2)][BPh(4)] (6), respectively. Similar reactions of 4A with organolithium reagents leads to intractable solids, presumably due to overreduction of the chromium(III) center. X-ray crystallographic analysis of 4A, 5, and 6 confirms that each possesses a largely undistorted octahedral chromium center, consistent with the observed S = (3)/(2) electronic ground states. Compounds 1, 2A, 2B, 4A, 4B, 5, and 6 are all active polymerization catalysts in the presence of methylalumoxane, producing low to moderate molecular weight high density polyethylene. PMID- 14552640 TI - New luminescent cyclometalated iridium(III) diimine complexes as biological labeling reagents. AB - We report the synthesis, characterization, and photophysical and electrochemical properties of thirty luminescent cyclometalated iridium(III) diimine complexes [Ir(N-C)(2)(N-N)](PF(6)) (HN-C = 2-phenylpyridine, Hppy; 2-(4 methylphenyl)pyridine, Hmppy; 3-methyl-1-phenylpyrazole, Hmppz; 7,8 benzoquinoline, Hbzq; 2-phenylquinoline, Hpq; N-N = 4-amino-2,2'-bipyridine, bpy NH(2); 4-isothiocyanato-2,2'-bipyridine, bpy-ITC; 4-iodoacetamido-2,2' bipyridine, bpy-IAA; 5-amino-1,10-phenanthroline, phen-NH(2); 5-isothiocyanato 1,10-phenanthroline, phen-ITC; 5-iodoacetamido-1,10-phenanthroline, phen-IAA). The X-ray crystal structure of [Ir(mppz)(2)(bpy-NH(2))](PF(6)) has also been investigated. Upon irradiation, all the complexes display intense and long-lived luminescence under ambient conditions and in 77-K glass. On the basis of the photophysical and electrochemical data, the emission of most of these complexes is assigned to an excited state of predominantly triplet metal-to-ligand charge transfer ((3)MLCT) (dpi(Ir) --> pi(N-N)) character. In some cases, triplet intraligand ((3)IL) (pi --> pi)(N-N or N-C(-)) excited states have also been identified. In view of the specific reactivity of the isothiocyanate and iodoacetamide moieties toward the primary amine and sulfhydryl groups, respectively, we have labeled various biological molecules with a selection of these luminescent iridium(III) complexes. The photophysical properties of the luminescent conjugates have been investigated. In addition, a heterogeneous assay for digoxin has also been designed on the basis of the recognition of biotinylated anti-digoxin by avidin labeled with one of the luminescent iridium(III) complexes. PMID- 14552641 TI - 4,4'-dithiodipyridine as a bridging ligand in osmium and ruthenium complexes: the electron conductor ability of the -S-S- bridge. AB - The compounds [Ru(NH(3))(5)(dtdp)](TFMS)(3), [Os(NH(3))(5)(dtdp)](TFMS)(3), [(NH(3))(5)Os(dtdp)Os(NH(3))(5)](TFMS)(6), [(NH(3))(5)Os(dtdp)Ru(NH(3))(5)](TFMS)(3)(PF(6))(2), and [(NH(3))(5)Os(dtdp)Fe(CN)(5)] (dtdp = 4,4'-dithiodipyridine, TFMS = trifluoromethanesulfonate) have been synthesized and characterized by elemental analysis, cyclic voltammetry, electronic, vibrational, EPR, and (1)H NMR spectroscopies. Changes in the electronic and voltammetric spectra of the ion complex [Os(NH(3))(5)(dtdp)](3+) as a function of the solution pH enable us to calculate the pK(a) for the [Os(NH(3))(5)(dtdpH)](4+) and [Os(NH(3))(5)(dtdpH)](3+) acids as 3.5 and 5.5, respectively. The comparison of the above pK(a) data with that for the free ligand (pK(1) = 4.8) provides evidence for the -S-S- bridge efficiency as an electron conductor between the two pyridine rings. The symmetric complex, [(NH(3))(5)Os(dtdp)Os(NH(3))(5)](6+), is found to exist in two geometric forms, and the most abundant form (most probably trans) has a strong conductivity through the -S-S- bridge, as is shown by EPR, which finds it to have an S = 1 spin state with a spin-spin interaction parameter of 150-200 G both in the solid sate and in frozen solution. Further the NMR of the same complex shows a large displacement of unpaired spin into the pi orbitals of the dttp ligand relative to that found in [Os(NH(3))(5)(dtdp)](3+). The comproportionation constant, K(c) = 2.0 x 10(5), for the equilibrium equation [Os(II)Os(II)] + [Os(III)Os(III)] right harpoon over left harpoon 2[Os(II)Os(III)] and the near-infrared band energy for the mixed-valence species (MMCT), [(NH(3))(5)Os(dtdp)Os(NH(3))(5)](5+) (lambda(MMCT) = 1665 nm, epsilon = 3.5 x 10(3) M(-)(1) cm(-)(1), deltanu(1/2) = 3.7 x 10(3) cm(-)(1), alpha = 0.13, and H(AB) = 7.8 x 10(2) cm(-)(1)), are quite indicative of strong electron delocalization between the two osmium centers. The electrochemical and spectroscopic data for the unsymmetrical binuclear complexes [(NH(3))(5)Os(III)(dtdp)Ru(II)(NH(3))(5)](5+) (lambda(MMCT) = 965 nm, epsilon = 2.2 x 10(2) M(-)(1) cm(-)(1), deltanu(1/2) = 3.0 x 10(3) cm(-)(1), and H(AB) = 2.2 x 10(2) cm(-)(1)) and [(NH(3))(5)Os(III)(dtdp)Fe(II)(CN)(5)] (lambda(MMCT) = 790 nm, epsilon = 7.5 x 10 M(-)(1) cm(-)(1), deltanu(1/2) = 5.4 x 10(3) cm(-)(1), and H(AB) = 2.0 x 10(2) cm(-)(1)) also suggest a considerable electron delocalization through the S-S bridge. As indicated by a comparison of K(c) and energy of the MMCT process in the iron, ruthenium, and osmium complexes, the electron delocalization between the two metal centers increases in the following order: Fe < Ru < Os. PMID- 14552642 TI - Kinetics and mechanism of formation of the platinum-thallium bond: the [(CN)(5)Pt Tl(CN)(3)](3)(-) complex. AB - Formation kinetics of the metal-metal bonded [(CN)(5)PtTl(CN)(3)](3)(-) complex from Pt(CN)(4)(2)(-) and Tl(CN)(4)(-) has been studied in the pH range of 5-10, using standard mix-and-measure spectrophotometric technique at pH 5-8 and stopped flow method at pH > 8. The overall order of the reaction, Pt(CN)(4)(2)(-) + Tl(CN)(4)(-) right harpoon over left harpoon [(CN)(5)PtTl(CN)(3)](3)(-), is 2 in the slightly acidic region and 3 in the alkaline region, which means first order for the two reactants in both cases and also for CN(-) at high pH. The two-term rate law corresponds to two different pathways via the Tl(CN)(3) and Tl(CN)(4)(-) complexes in acidic and alkaline solution, respectively. The two complexes are in fast equilibrium, and their actual concentration ratio is controlled by the concentration of free cyanide ion. The following expression was derived for the pseudo-first-order rate constant of the overall reaction: k(obs) = (k(1)(a)[Tl(CN)(4)(-) + (k(1)(a)/K(f)))(1/(1 + K(p)[H(+)]))[CN(-)](free) + k(1)(b)[Tl(CN)(4)(-)] + (k(1)(b)/K(f)), where k(1)(a) and k(1)(b) are the forward rate constants for the alkaline and slightly acidic paths, K(f) is the stability constant of [(CN)(5)PtTl(CN)(3)](3)(-), and K(p) is the protonation constant of cyanide ion. k(1)(a) = 143 +/- 13 M(-)(2) s(-)(1), k(1)(b) = 0.056 +/- 0.004 M( )(1) s(-)(1), K(f) = 250 +/- 54 M(-)(1), and log K(p) = 9.15 +/- 0.05 (I = 1 M NaClO(4), T = 298 K). Two possible mechanisms were postulated for the overall reaction in both pH regions, which include a metal-metal bond formation step and the coordination of the axial cyanide ion to the platinum center. The alternative mechanisms are different in the sequence of these steps. PMID- 14552643 TI - Synthesis and structures of nickel and palladium salicylaldiminato 1,3,5-triaza-7 phosphaadamantane (PTA) complexes. AB - The synthesis of nickel(II) and palladium(II) salicylaldiminato complexes incorporating the water-soluble phosphine 1,3,5-triaza-7-phosphaadamantane(PTA) has been achieved employing two preparative routes. Reaction of the original ethylene polymerization catalyst developed by Grubbs and co-workers (Organometallics 1998, 17, 3149), (salicylaldiminato)Ni(Ph)PPh(3), with PTA using a homogeneous methanol/toluene solvent system resulted in the formation of the PTA analogues in good yields. Alternatively, complexes of this type may be synthesized via a direct approach utilizing (tmeda)M(CH(3))(2) (M = Ni, Pd), the corresponding salicylaldimine, and PTA. Yields by this method were generally near quantitative. The complexes were characterized in solution by (1)H/(13)C/(31)P NMR spectroscopy and in the solid-state by X-ray crystallography. All derivatives exhibited square-planar geometry with the bulky isopropyl groups on the aniline being perpendicular to the plane formed by the metal center and its four ligands. Such orientation of these sterically encumbering groups is responsible for polymer chain growth during olefin polymerization in favor of chain termination via beta-hydride elimination. Polymerization reactions were attempted using the nickel-PTA complexes in a biphasic toluene/water mixture in an effort to initiate ethylene polymerization by trapping the dissociated phosphine ligand in the water layer, thereby eliminating the need for a phosphine scavenger. Unfortunately, because of the strong binding ability of the small, donating phosphine(PTA) as compared to PPh(3), phosphine dissociation did not occur at a temperature where the complexes are thermally stable. PMID- 14552644 TI - The kinetics and mechanism of the ferrate(VI) oxidation of hydroxylamines. AB - Aqueous solutions of potassium ferrate(VI) cleanly and rapidly oxidize hydroxylamine to nitrous oxide, N-methylhydroxylamine to nitrosomethane, N phenylhydroxylamine to nitrosobenzene, and O-methylhydroxylamine to methanol and nitrogen. The kinetics show first-order behavior with respect to each reactant and a two term component representing acid dependent and independent pathways. A general mechanism involving intermediate formation coupled with a two-electron oxidation is proposed. PMID- 14552646 TI - Valence compounds versus metals. Synthesis, characterization, and electronic structures of cubic Ae(4)Pn(3) phases in the systems Ae = Ca, Sr, Ba, Eu; Pn = As, Sb, Bi. AB - The isostructural compounds Sr(4)Bi(3), Ba(4)Bi(3), and Ba(4)As( approximately )(2.60) were prepared by direct reactions of the corresponding elements and their structures determined from single-crystal X-ray diffraction data as anti Th(3)P(4) type in the cubic space group I43d, Z = 4 (a = 10.101(1) A, 10.550(1) A, 9.973 (1) A, respectively). The two bismuth compounds are stoichiometric, and the arsenide refines as Ba(4)As(2.60(2)). Only unrelated phases are obtained for all binary combinations among the title components for either Ca or Sb. The magnetic susceptibility and resistivities of Ba(4)Bi(3) and Eu(4)Bi(3) show that they are good metallic conductors ( approximately 40 microomega.cm at 298 K), whereas Ba(4)As(2.60) exhibits rho(150) > 1000 microomega.cm. The electronic structures of Sr(4)Bi(3), Ba(4)Bi(3), and Ba(4)As(3) were calculated by TB-LMTO ASA methods. Mixing of cation d states into somewhat disperse valence p bands on Bi results in empty bands at E(F) and metallic behavior, whereas the narrower valence band in the electron-deficient Ba(4)As(3) leads to vacancies in about 11% of the anion sites and a valence compound. PMID- 14552645 TI - Coordination properties of new bis(1,4,7-triazacyclononane) ligands: a highly active dizinc complex in phosphate diester hydrolysis. AB - The synthesis and characterization of three new bis([9]aneN(3)) ligands, containing respectively 2,2'-bipyridine (L(1)), 1,10-phenanthroline (L(2)), and quinoxaline (L(3)) moieties linking the two macrocyclic units, are reported. Proton binding and Cu(II), Zn(II), Cd(II), and Pb(II) coordination with L(1)-L(3) have been studied by potentiometric titrations and, for L(1) and L(2), by spectrophotometric UV-vis measurements in aqueous solutions. All ligands can give stable mono- and dinuclear complexes. In the case of L(1), trinuclear Cu(II) complexes are also formed. The stability constants and structural features of the formed complexes are strongly affected by the different architecture and binding properties of the spacers bridging the two [9]aneN(3) units. In the case of the L(1) and L(2) mononuclear complexes, the metal is coordinated by the three donors of one [9]aneN(3) moiety; in the [ML(2)](2+) complexes, however, the phenanthroline nitrogens are also involved in metal binding. Finally, in the [ML(3)](2+) complexes both macrocyclic units, at a short distance from each other, can be involved in metal coordination, giving rise to sandwich complexes. In the binuclear complexes each metal ion is generally coordinated by one [9]aneN(3) unit. In L(1), however, the dipyridine nitrogens can also act as a potential binding site for metals. The dinuclear complexes show a marked tendency to form mono-, di-, and, in some cases, trihydroxo species in aqueous solutions. The resulting M-OH functions may behave as nucleophiles in hydrolytic reactions. The hydrolysis rate of bis(p-nitrophenyl)phosphate (BNPP) was measured in aqueous solution at 308.1 K in the presence of the L(2) and L(3) dinuclear Zn(II) complexes. Both the L(2) complexes [Zn(2)L(2)(OH)(2)](2+) and [Zn(2)L(2)(OH)(3)](+) and the L(3) complex [Zn(2)L(3)(OH)(3)](+) promote BNPP hydrolysis. The [Zn(2)L(3)(OH)(3)](+) complex is ca. 2 orders of magnitude more active than the L(2) complexes, due both to the short distance between the metal centers in [Zn(2)L(3)(OH)(3)](+), which could allow a bridging interaction of the phosphate ester, and to the simultaneous presence of single-metal bound nucleophilic Zn-OH functions. These structural features are substantially corroborated by semiempirical PM3 calculations carried out on the mono-, di-, and trihydroxo species of the L(3) dizinc complex. PMID- 14552647 TI - A schiff-base bibracchial lariat ether forming a cryptand-like cavity for lanthanide ions. AB - We report here a structural and photophysical study of lanthanide(III) complexes with the di-deprotonated form of the bibracchial lariat ether N,N'-bis(2 salicylaldiminobenzyl)-1,10-diaza-15-crown-5. The X-ray crystal structures of [Ce(L(2)-2H)](ClO(4)).0.5H(2)O (2) and [Sm(L(2)-2H)](ClO(4)).C(3)H(8)O (5b) show the metal ion being nine-coordinated and deeply buried in the cavity of the dianionic receptor. Thanks to the formation of a pseudomacrocycle through pi-pi interaction between one of the phenol rings and one of the benzyl rings, the complexes present a cryptand-like structure in the solid state. (1)H and (13)C NMR studies on the La(III) complex point that the solid state structure is essentially maintained in acetonitrile solution. High-resolution laser-excited emission spectra of the crystalline Eu(III) complex demonstrate the presence of several coordination sites arising from different conformations of the crown moiety. The ligand-to-Eu(III) energy transfer is relatively efficient at low temperature, but back transfer is implied in the deactivation process, especially at room temperature, because the ligand triplet state lies at very low energy. However, the low energy of the (3)pipi state provides an efficient conversion of the visible light absorbed into near-infrared light emitted by the Nd(III) ion. PMID- 14552651 TI - Differences in mechanism of action between current and future antidepressants. AB - Antidepressants are divided into several classes on the basis of their pharmacologic mechanisms of action, which are thought to be responsible for both their therapeutic actions and their side effect profiles. All classes currently available in the United States affect serotonin, norepinephrine, and/or dopamine neurotransmission. New agents in development also affect neurotransmission of such monoamines and include serotonin-norepinephrine reuptake inhibitors, serotonin-selective agents, selective monoamine oxidase inhibitors, and selective norepinephrine reuptake inhibitors. Treatments with entirely new mechanisms of action are also being studied, including hormone-linked treatments such as estrogen replacement therapy and the steroid antagonist mifepristone (RU-486 or C 1073); novel antagonist peptides such as corticotropin-releasing factor, neurokinins, and injectable pentapeptides; and agents that affect glutamate neurotransmission. The introduction of antidepressants with novel mechanisms of action could potentially revolutionize the treatment of depression. PMID- 14552650 TI - Interactions of antidepressants with neurotransmitter transporters and receptors and their clinical relevance. AB - This review discusses the pharmacology of antidepressants as it relates to their blockade of human transporters for norepinephrine, serotonin, and dopamine, as well as their blockade of several different human neurotransmitter receptors. This blockade by antidepressants of transporters and receptors is thought to relate to therapeutic and adverse effects of these compounds. Knowledge of the preclinical pharmacology of antidepressants will help the clinician to identify adverse effects and drug interactions that may occur with these agents. The article also reviews drugs that have dual action (effects on serotonin and norepinephrine), which may provide a therapeutic advantage in treating depressed patients. PMID- 14552652 TI - Evaluating antidepressant therapies: remission as the optimal outcome. AB - Depression is the fourth-largest contributor to the global burden of disease, and it causes profound suffering and extreme costs to health care systems and society. Although there have been many new antidepressants introduced, few depressed individuals receive the optimal treatment. One problem is that the traditional definition of response to antidepressant therapy, i.e., a 50% improvement in symptoms, ensures little beyond a reduction of syndromal intensity. Responders who have persistent depressive symptoms experience ongoing psychosocial dysfunction, poorer health, and an increased risk of relapse. The goal of the first or acute-phase treatment should be complete remission of symptoms and a full return to premorbid levels of functioning. Remission is also a necessary, transitional state toward sustained recovery. Within this context, evidence pertaining to various treatment approaches is reexamined, taking into account critical methodological issues such as design sensitivity and statistical power. Whereas results of individual studies are inconsistent, the findings of meta-analyses (i.e., quantitative and pooled) suggest that both psychotherapy pharmacotherapy combinations and use of antidepressants that enhance serotonergic and noradrenergic neurotransmission increase the likelihood of remission. PMID- 14552653 TI - The role of the serotonergic and noradrenergic neurotransmitter systems in the treatment of psychological and physical symptoms of depression. AB - Major depressive disorder is a medical condition that includes abnormalities of affect and mood, cognition, and physical functioning. In particular, as many as 76% of patients suffering from depression are found to report somatic symptoms, including various types of pain such as headaches, stomach pain, back pain, and vague, poorly localized pain. Although the pathophysiology of depression is still unknown, there is significant evidence for abnormalities of the norepinephrine (NE) and serotonin (5-HT) neurotransmitter systems in depressive disorders. Interestingly, both 5-HT and NE also appear to exert analgesic effects via descending pain pathways and therefore play a modulating role in pain. There are many effective antidepressant treatments available. However, residual symptoms are relatively common, among both partial responders and responders without remission. A recent study from our group has shown that responders who have not achieved remission have significantly more somatic symptoms than remitters following 8 weeks of treatment with fluoxetine. These data may suggest that antidepressants that are particularly effective in the treatment of pain and painful physical symptoms may yield higher remission rates in major depressive disorder. PMID- 14552654 TI - Efficacy and tolerability of duloxetine, a novel dual reuptake inhibitor, in the treatment of major depressive disorder. AB - Although highly selective antidepressants such as the selective serotonin reuptake inhibitors represent an advance over older drugs with respect to tolerability, they are not more effective than previous agents. Antidepressants that enhance transmission in more than one monoamine system may have greater efficacy than highly selective drugs, while equaling or improving their adverse effect profiles. This article reviews the properties of duloxetine, a potent and balanced inhibitor of norepinephrine and serotonin reuptake. Controlled studies indicate a high degree of efficacy, tolerability, and safety for duloxetine in the treatment of major depressive disorder. In particular, rapid therapeutic onset and high remission rates have been noted. Duloxetine appears to have significant benefit in the treatment of the painful physical symptoms associated with depression. The continued presence of such symptoms may predict relapse. Accordingly, it is hoped that duloxetine therapy may reduce the likelihood of depressive relapse. PMID- 14552656 TI - Real-time PCR based on SYBR-Green I fluorescence: an alternative to the TaqMan assay for a relative quantification of gene rearrangements, gene amplifications and micro gene deletions. AB - BACKGROUND: Real-time PCR is increasingly being adopted for RNA quantification and genetic analysis. At present the most popular real-time PCR assay is based on the hybridisation of a dual-labelled probe to the PCR product, and the development of a signal by loss of fluorescence quenching as PCR degrades the probe. Though this so-called 'TaqMan' approach has proved easy to optimise in practice, the dual-labelled probes are relatively expensive. RESULTS: We have designed a new assay based on SYBR-Green I binding that is quick, reliable, easily optimised and compares well with the published assay. Here we demonstrate its general applicability by measuring copy number in three different genetic contexts; the quantification of a gene rearrangement (T-cell receptor excision circles (TREC) in peripheral blood mononuclear cells); the detection and quantification of GLI, MYC-C and MYC-N gene amplification in cell lines and cancer biopsies; and detection of deletions in the OPA1 gene in dominant optic atrophy. CONCLUSION: Our assay has important clinical applications, providing accurate diagnostic results in less time, from less biopsy material and at less cost than assays currently employed such as FISH or Southern blotting. PMID- 14552657 TI - Trustworthiness and metrics in visualizing similarity of gene expression. AB - BACKGROUND: Conventionally, the first step in analyzing the large and high dimensional data sets measured by microarrays is visual exploration. Dendrograms of hierarchical clustering, self-organizing maps (SOMs), and multidimensional scaling have been used to visualize similarity relationships of data samples. We address two central properties of the methods: (i) Are the visualizations trustworthy, i.e., if two samples are visualized to be similar, are they really similar? (ii) The metric. The measure of similarity determines the result; we propose using a new learning metrics principle to derive a metric from interrelationships among data sets. RESULTS: The trustworthiness of hierarchical clustering, multidimensional scaling, and the self-organizing map were compared in visualizing similarity relationships among gene expression profiles. The self organizing map was the best except that hierarchical clustering was the most trustworthy for the most similar profiles. Trustworthiness can be further increased by treating separately those genes for which the visualization is least trustworthy. We then proceed to improve the metric. The distance measure between the expression profiles is adjusted to measure differences relevant to functional classes of the genes. The genes for which the new metric is the most different from the usual correlation metric are listed and visualized with one of the visualization methods, the self-organizing map, computed in the new metric. CONCLUSIONS: The conjecture from the methodological results is that the self organizing map can be recommended to complement the usual hierarchical clustering for visualizing and exploring gene expression data. Discarding the least trustworthy samples and improving the metric still improves it. PMID- 14552659 TI - Discrepancy between cardiorespiratory system and skeletal muscle in elite cyclists after hypoxic training. AB - BACKGROUND: The purpose of this study was to determine the effects of hypoxic training on the cardiorespiratory system and skeletal muscle among well-trained endurance athletes in a randomized cross-over design. METHODS: Eight junior national level competitive cyclists were separated into two groups; Group A trained under normoxic condition (21% O2) for 2 hours/day, 3 days/week for 3 weeks while Group B used the same training protocol under hypoxic condition (15% O2). After 3 weeks of each initial training condition, five weeks of self training under usual field conditions intervened before the training condition was switched from NT to HT in Group A, from HT to NT in Group B. The subjects were tested at sea level before and after each training period. O2 uptake (O2), blood samples, and muscle deoxygenation were measured during bicycle exercise test. RESULTS AND DISCUSSION: No changes in maximal workload, arterial O2 content, O2 at lactate threshold and O2max were observed before or after each training period. In contrast, deoxygenation change during submaximal exercise in the vastus lateralis was significantly higher at HT than NT (p < 0.01). In addition, half time of oxygenation recovery was significantly faster after HT (13.2 PlusMinus; 2.6 sec) than NT (18.8 PlusMinus; 2.7 sec) (p < 0.001). CONCLUSIONS: Three weeks of HT may not give an additional performance benefit at sea level for elite competitive cyclists, even though HT may induce some physiological adaptations on muscle tissue level. PMID- 14552660 TI - Fine structure of the low-frequency spectra of heart rate and blood pressure. AB - BACKGROUND: The aim of this study was to explore the principal frequency components of the heart rate and blood pressure variability in the low frequency (LF) and very low frequency (VLF) band. The spectral composition of the R-R interval (RRI) and systolic arterial blood pressure (SAP) in the frequency range below 0.15 Hz were carefully analyzed using three different spectral methods: Fast Fourier transform (FFT), Wigner-Ville distribution (WVD), and autoregression (AR). All spectral methods were used to create time-frequency plots to uncover the principal spectral components that are least dependent on time. The accurate frequencies of these components were calculated from the pole decomposition of the AR spectral density after determining the optimal model order--the most crucial factor when using this method--with the help of FFT and WVD methods. RESULTS: Spectral analysis of the RRI and SAP of 12 healthy subjects revealed that there are always at least three spectral components below 0.15 Hz. The three principal frequency components are 0.026 +/- 0.003 (mean +/- SD) Hz, 0.076 +/- 0.012 Hz, and 0.117 +/- 0.016 Hz. These principal components vary only slightly over time. FFT-based coherence and phase-function analysis suggests that the second and third components are related to the baroreflex control of blood pressure, since the phase difference between SAP and RRI was negative and almost constant, whereas the origin of the first component is different since no clear SAP-RRI phase relationship was found. CONCLUSION: The above data indicate that spontaneous fluctuations in heart rate and blood pressure within the standard low frequency range of 0.04-0.15 Hz typically occur at two frequency components rather than only at one as widely believed, and these components are not harmonically related. This new observation in humans can help explain divergent results in the literature concerning spontaneous low-frequency oscillations. It also raises methodological and computational questions regarding the usability and validity of the low-frequency spectral band when estimating sympathetic activity and baroreflex gain. PMID- 14552658 TI - OXBench: a benchmark for evaluation of protein multiple sequence alignment accuracy. AB - BACKGROUND: The alignment of two or more protein sequences provides a powerful guide in the prediction of the protein structure and in identifying key functional residues, however, the utility of any prediction is completely dependent on the accuracy of the alignment. In this paper we describe a suite of reference alignments derived from the comparison of protein three-dimensional structures together with evaluation measures and software that allow automatically generated alignments to be benchmarked. We test the OXBench benchmark suite on alignments generated by the AMPS multiple alignment method, then apply the suite to compare eight different multiple alignment algorithms. The benchmark shows the current state-of-the art for alignment accuracy and provides a baseline against which new alignment algorithms may be judged. RESULTS: The simple hierarchical multiple alignment algorithm, AMPS, performed as well as or better than more modern methods such as CLUSTALW once the PAM250 pair score matrix was replaced by a BLOSUM series matrix. AMPS gave an accuracy in Structurally Conserved Regions (SCRs) of 89.9% over a set of 672 alignments. The T-COFFEE method on a data set of families with <8 sequences gave 91.4% accuracy, significantly better than CLUSTALW (88.9%) and all other methods considered here. The complete suite is available from http://www.compbio.dundee.ac.uk. CONCLUSIONS: The OXBench suite of reference alignments, evaluation software and results database provide a convenient method to assess progress in sequence alignment techniques. Evaluation measures that were dependent on comparison to a reference alignment were found to give good discrimination between methods. The STAMP Sc Score which is independent of a reference alignment also gave good discrimination. Application of OXBench in this paper shows that with the exception of T-COFFEE, the majority of the improvement in alignment accuracy seen since 1985 stems from improved pair-score matrices rather than algorithmic refinements. The maximum theoretical alignment accuracy obtained by pooling results over all methods was 94.5% with 52.5% accuracy for alignments in the 0-10 percentage identity range. This suggests that further improvements in accuracy will be possible in the future. PMID- 14552661 TI - Consequences of malnutrition on human capital and productivity. PMID- 14552662 TI - Out of the box. PMID- 14552663 TI - Nutrition as a preventive strategy against adverse maternal pregnancy outcomes - a USAID/Wellcome Trust Workshop at Merton College, Oxford, UK, 18-19 July 2002. PMID- 14552664 TI - Secondary data analyses of dietary surveys undertaken in South Africa to determine usual food consumption of the population. AB - OBJECTIVE: The primary objective of this study was to generate a reference table of food items and average amounts of these items consumed by South Africans, for the Department of Health. The reference table was required to be representative of foods and beverages eaten frequently by children and adults from all age and ethnic groups in order for the Department of Health to test for contaminants in these foods. DESIGN: The National Food Consumption Survey (NFCS) served as a framework for compiling data on children since this was a national representative survey of 1-9-year-old children undertaken in South Africa in 1999. However, there has never been a national dietary survey on adults in South Africa. Consequently the data had to be extrapolated from existing isolated surveys on adults. Secondary data analysis was conducted on existing dietary databases (raw data) obtained from surveys undertaken on adults in South Africa between 1983 and 2000. Available datasets were regional and independent, and were not individually representative of the South African diet. It was therefore necessary to use different statistical methods, including factor analyses, weighting and correlations, to generate ethnic and geographic representative data for adults. Two methods were used: Method 1, which corresponded with results of the NFCS (over-sampled for low socio-economic status), and Method 2, which was based on ethnic proportions of the population. RESULTS: The secondary data analyses generated food items most commonly consumed by the South African adult population (Method 1) in descending frequency of usage and average (mean) amount per day: maize porridge (78%/848 g), white sugar (77%/27 g), tea (68%/456 g), brown bread (55%/165 g), white bread (28%/163 g), non-dairy creamer (25%/6 g), brick margarine (21%/19 g), chicken meat (19%/111 g), full-cream milk (19%/204 g) and green leaves (17%/182 g). In 6-9-year-olds, maize porridge (72%/426 g), sugar (76%/23 g), tea (51%/258 g), full-cream milk (35%/171 g) and white bread (33%/119 g) were eaten most frequently. Similarly, in 1-5-year-olds, the foods consumed most frequently were maize porridge (80%/426 g), sugar (76%/21 g), tea (44%/224 g), full-cream milk (39%/186 g) and white bread (24%/83 g). In order to evaluate the validity of the adult data generated, kilojoule values of the individual food items (per capita) were compared with food balance sheets (FBSs). The comparison was favourable except that the FBSs had a higher overall energy intake per capita of between 22 and 28%. CONCLUSION: Reference tables of commonly consumed foods and beverages were generated at minimal cost based on secondary data analyses of past dietary surveys in different South African populations. PMID- 14552665 TI - Risk factors for undernutrition of young children in a rural area of South Africa. AB - OBJECTIVE: To identify the factors associated with childhood undernutrition. DESIGN: Cross-sectional survey. SETTING: Hlabisa health district in KwaZulu/Natal, South Africa. SUBJECTS: Eight hundred and sixty-eight children aged 3-59 months. METHODS: Questionnaire survey and anthropometric survey of 516 random households with children in the health district. Multivariate analysis took into account the hierarchical relationships between the proposed risk factors. This conceptual model was built up during qualitative studies and with reference to international research in this area. RESULTS: The mean Z-scores for weight-for-age and height-for-age were -0.52 (95% confidence interval (CI) -0.44 to -0.60) and -1.25 (95% CI -1.15 to -1.35), respectively. Of the children, 26.3% (95% CI 23.3-29.3%) were stunted, 12.0% (95% CI 9.8-4.2%) were underweight-for age (UWFA) and only 1.3% were wasted. Migrant father, mother's education, literate mother, whether the house was made of traditional materials, the presence of a toilet, whether the last child was breast-fed, duration of breast feeding and birth weight were found to be significantly related to UWFA, the latter with an odds ratio of more than eight. CONCLUSION: While there are individual health and feeding behaviours that need to be addressed, this study has also uncovered the maternal and socio-economic factors which present an array of constraints in the social environment shaping the infant feeding and caring practices employed by women. For the full potential of nutrition promotion activities to be realised, there is a need to address some of these 'deeper' constraints. PMID- 14552666 TI - Obesity among children attending elementary public schools in Sao Paulo, Brazil: a case--control study. AB - OBJECTIVES: To describe obesity among students of public schools in Sao Paulo and to identify risk factors for this nutritional and physical activity disorder. DESIGN: Case-control study of obese and non-obese schoolchildren to study risk factors for obesity. SETTING: Anthropometric survey including 2519 children attending eight elementary public schools in Sao Paulo, Brazil. SUBJECTS: Schoolchildren aged 7-10 years, of whom 223 were obese (cases; weight-for-height greater than or equal to two standard deviations (>or=2SD) above the median of the National Center for Health Statistics (NCHS) reference population) and 223 were eutrophic (controls; weight-for-height +/-1SD from NCHS median). MEASUREMENTS: Parents or guardians of the 446 cases and controls were interviewed about the children's eating behaviours and habits. RESULTS: The prevalence of obesity (weight-for-height >or=2SD) in the surveyed population was 10.5%. A logistic regression model fitted to the case-control dataset showed that obesity was positively associated with the following factors: birth weight >or=3500 g (odds ratio (OR) 1.83, 95% confidence interval (CI) 1.21-2.78), child's appetite at meals (OR 3.81, 95% CI 2.49-5.83), watching television for 4 h per day or longer (OR 2.07, 95% CI 1.32-3.24), mother's schooling >4 years (OR 1.85, 95% CI 1.25-2.75) and parents' body mass index >or=30 kg x m(-2) (OR 2.50, 95% CI 1.43 4.37). CONCLUSIONS: The explanatory multivariate model points to preventive measures that would encourage knowledge of the children and their guardians in relation to a balanced diet and a less sedentary lifestyle, such as reducing television viewing. Schoolchildren with a birth weight of 3500 g or more or whose parents are obese should receive special attention in the prevention of obesity. PMID- 14552667 TI - Socio-economic, dietary, activity, nutrition and body weight transitions in South Korea. AB - OBJECTIVE: Socio-economic development influences many factors that affect health, especially diet and nutrition. This investigation proposes that a system of transitions occur as societies develop, with socio-economic, physical activity, dietary, nutrition and body weight transitions operating in relationship with each other. This model of transitions was examined empirically using South Korea as an example of a nation that has undergone considerable changes. DESIGN: Data were drawn from published government reports: the Korean National Nutrition Survey and annual reports at the national level for the years between 1969 and 1993. The socio-economic transition was assessed by gross national product. The physical activity transition was assessed using annual proportions of the population involved in primary, secondary and tertiary industries, as well as the number of cars and driver's licences. The dietary transition was measured by plant and animal food consumption. The nutrition transition was assessed by percentages of energy from carbohydrate, protein and fat. The body weight transition was measured by body mass index calculated from the average height and weight of adolescents. RESULTS: Results revealed that the transitions were highly correlated as expected, with the socio-economic transition exhibiting major changes. South Koreans tended to decrease their physical activity and plant food consumption, and to increase animal food consumption, percentage of energy from dietary fat and body weight, in relationship to the socio-economic transition. CONCLUSION: Examining a system of transitions on a national level in one country that has undergone rapid economic development may provide a strategy for examining how such transitions operate in other nations. PMID- 14552668 TI - Relationship of tobacco smoking with serum vitamin B12, folic acid and haematological indices in healthy adults. AB - OBJECTIVES: To investigate the effects of tobacco smoking on serum vitamin B12, folic acid and haematological parameters in healthy Thai smokers and non-smokers. DESIGN: Cross-sectional study of smokers and non-smokers in a military unit in Bangkok, Thailand. SETTING: A military unit in Thailand. SUBJECTS: One hundred and twenty-three male smokers from a military unit in Bangkok, who participated voluntarily in the study, were investigated. Sixty-six male non-smokers from the same unit were selected as controls. Fasting blood samples were collected for investigation of vitamin B12, folic acid and haematological variables. RESULTS: The serum folic acid concentration of smokers was lower than that of non-smokers, but was not statistically significantly different. Haemoglobin was lower in smokers than in non-smokers; 16.3% of smokers were anaemic compared with only 3.0% of non-smokers. Anaemia was not related to folate deficiency. The white blood cell count was found to be higher in smokers than in non-smokers. CONCLUSION: The results of this study suggest that there were low serum folic acid concentrations in smokers compared with non-smokers, which might contribute to the development of vascular and cardiovascular diseases. The higher white blood cell count might be indicative alterations in the immune functions of smokers. PMID- 14552669 TI - A qualitative assessment of nutrition knowledge levels and dietary intake of schoolchildren in Hyderabad. AB - OBJECTIVES: To assess the nutrition knowledge levels and dietary intake pattern of schoolchildren belonging to two groups of different socio-economic status (SES; high income/high SES and low income/low SES). DESIGN: A purposive sampling method was employed. A validated food-frequency questionnaire was administered to assess the dietary intake of schoolchildren in four schools from two different socio-economic strata in the month of January 2001. The children were divided into two groups, one serving as the experimental group and the other as the control group. SUBJECTS: Two hundred and seventy-two children aged between 12 and 14 years. RESULTS: There was a significant improvement (P<0.001) in the knowledge levels of high-SES schoolchildren as compared with low-SES schoolchildren. A significant difference was observed in the intake of protective foods like milk and milk products, green leafy vegetables and fruits between the two income groups. However, children from the high SES background preferred fast foods such as noodles and corn flakes to traditional foods. Irrespective of income group, most of the children consumed carbonated beverages. CONCLUSIONS: There was a significant difference in the intakes of protective foods and fast foods between the different income groups. However, the increased intake of fast foods and carbonated beverages by the children irrespective of SES needs to be discouraged as a part of nutrition education. The study indicated the need for repeated interventions for improvement of nutrition knowledge levels in low-SES children. PMID- 14552670 TI - Association of education with dietary intake among young adults in the bi-ethnic Coronary Artery Risk Development in Young Adults (CARDIA) cohort. AB - OBJECTIVE: To examine associations of changes in dietary intake with education in young black and white men and women. DESIGN: The Coronary Artery Risk Development in Young Adults (CARDIA) study, a multi-centre population-based prospective study. Dietary intake data at baseline and year 7 were obtained from an extensive nutritionist-administered diet history questionnaire with 700 items developed for CARDIA. SETTING: Participants were recruited in 1985-1986 from four sites: Birmingham, Alabama; Chicago, Illinois; Minneapolis, Minnesota; and Oakland, California. SUBJECTS: Participants were from a general community sample of 703 black men (BM), 1006 black women (BW), 963 white men (WM) and 1054 white women (WW) who were aged 18-30 years at baseline. Analyses here include data for baseline (1985-1986) and year 7 (1992-1993). RESULTS: Most changes in dietary intake were observed among those with high education (>or=12 years) at both examinations. There was a significant decrease in intake of energy from saturated fat and cholesterol and a significant increase in energy from starch for each race-gender group (P<0.001). Regardless of education, taste was considered an important influence on food choice. CONCLUSION: The inverse relationship of education with changes in saturated fat and cholesterol intakes suggests that national public health campaigns may have a greater impact among those with more education. PMID- 14552671 TI - Two approaches to derive a proposal for added sugars intake for German children and adolescents. AB - OBJECTIVE: Intake of sugars is restricted in most international food guides. However, in recent recommendations quantified limits of added sugars intake have not been given, although deemed necessary by those who criticised the recommendations. DESIGN: Two approaches to derive a scientifically based quantified limit of added sugars intake for German children and adolescents are suggested. For the first dietary survey approach, 5120 three-day weighed dietary records from the Dortmund Nutritional and Anthropometric Longitudinally Designed (DONALD) Study were used. Added sugars intakes in records of high vs. low dietary quality, measured by a nutrient intake score, were compared. For the second approach, a total dietary concept for German children and adolescents developed and evaluated by the Research Institute of Child Nutrition - the Optimised Mixed Diet - was used. RESULTS: Whereas in the latter dietary concept an added sugars intake of about 6% of energy intake is tolerated, the dietary survey approach resulted in only small differences between high and low dietary quality, with a median added sugars intake of 12% of energy in records with high dietary quality. CONCLUSIONS: A reasonable dietary quality is possible within higher ranges of added sugars intake than derived from the dietary concept approach. Therefore we suggest a range of intake of added sugars for German children and adolescents, from 6% to 12% of energy. PMID- 14552672 TI - Selected nutritional biomarkers predict diet quality. AB - OBJECTIVE: To examine associations of biomarkers of nutrient intake with overall diet quality. SUBJECTS: A convenience sample of 102 healthy postmenopausal women in Seattle, Washington (USA). DESIGN AND METHOD: Participants attended a study centre where they provided fasting blood specimens and completed a 122-item validated food-frequency questionnaire (FFQ). Data from the FFQ were used to calculate Diet Quality Index (DQI) scores and categorise women as having diets of excellent, good, fair or poor quality. The blood specimens were analysed for nine phospholipid fatty acids (as percentage of total) and serum concentrations of vitamin C, alpha-tocopherol, gamma-tocopherol, vitamin B12, folate and six carotenoids. Multivariate linear regression was used to model associations of the nutrient biomarkers with DQI scores. RESULTS: Compared with women with poor quality diets, women with excellent diets, as measured by the DQI, had higher plasma concentrations of vitamin C (P for trend=0.01), alpha-tocopherol (P for trend=0.02) and beta-cryptoxanthin (P for trend=0.03). Women with excellent diets also had lower proportions of plasma phospholipid fatty acids of two potentially atherogenic fatty acids: stearic acid (P for trend=0.01) and behenic acid (P for trend=0.03). A group of six biomarkers explained a moderate proportion of the total variability in DQI scores (36%). CONCLUSIONS: These objective measures of dietary intake support the use of the DQI as a useful tool to measure dietary patterns. PMID- 14552673 TI - The importance of composite foods for estimates of vegetable and fruit intakes. AB - OBJECTIVE: To evaluate the contribution of composite foods to vegetable and fruit intakes in Irish adults and to compliance with dietary guidelines for vegetable and fruit intake. DESIGN: Data were analysed from the North/South Ireland Food Consumption Survey of 18-64-year-old adults (n=1379; 662 men, 717 women), which used a 7-day food diary to estimate food intake. RESULTS: The mean intake of vegetables (excluding potatoes) was 140 g day(-1) (men 149 g day(-1); women 132 g day(-1)), of fruit was 136 g day(-1) (men 133 g day(-1); women 140 g day(-1)) and of potatoes was 227 g day(-1) (men 296 g day(-1); women 163 g day(-1)). The mean daily intakes of vegetables, fruit and potatoes from composite foods were 37 g (26%), 6 g (5%) and 17 g (7%), respectively. The mean intake of vegetables from composite foods was unrelated to age or gender, but increased with increasing social class and level of education attained. The proportions of men and women meeting the recommendation for >or=400 g day(-1) (5 servings of 80 g per day) of vegetables and fruit were 21% (15% excluding composite foods) and 19% (12% excluding composite foods), respectively. Compliance with the dietary recommendation decreased with decreasing levels of educational attainment and social class. CONCLUSION: Intakes of vegetables and fruit are low compared with current dietary recommendations, particularly in those of lower levels of educational attainment and social class. Composite foods are an important source of vegetables (less importantly of fruit) and should be included when estimating vegetable intakes. Failure to do so may result in bias in estimates of intake and of compliance with dietary guidelines for population groups, as well as misclassification of individuals by level of intake. PMID- 14552674 TI - [Dynamic observation IgG and IgM antibodies in patients with severe acute respiratory syndrome]. AB - OBJECTIVE: To observe IgG and IgM antibodies in patients with severe acute respiratory syndrome (SARS) and to explore their significance. METHODS: IgG and IgM antibodies were detected in SARS Patients in the first, recovery and follow up stages, the front healthy doctors and nurses and healthy volunteers with indirect enzyme-linked immunoadsorbent assay. RESULTS: The positive ratio of IgG was higher than that of IgM in SARS patients during the first stage. During the recovery, follow-up stages and in the front healthy crowd, the positive ratios of IgG were all higher than those of IgM. CONCLUSION: The changeable tendency SARS antibody is more different than that of other infectious diseases. This tendency can be used as a indicator for epidemiological study and serum diagnose. PMID- 14552675 TI - [Analysis and causation discussion of 185 severe acute respiratory syndrome dead cases]. AB - OBJECTIVE: To proceed the retrospective analysis of the dead reasons of patients with severe acute respiratory syndrome (SARS), in order to provide experience for decreasing mortality of SARS patients in clinic. METHODS: Retrospective analyze of 185 dead cases of SARS patients, including age, basic diseases, laboratory tests, radiological test and therapy. RESULTS: There were 102 men and 83 women in 185 dead cases of SARS, in which average age was 56.4 years, 85 cases were older than 60 years. 147 cases (79.5%) had basic diseases, cardiovascular disease (57 cases) was at the first Place; diabetes mellitus (41 cases) was at the second Place; cerebrovascular disease (19 cases) was at the third place; 15 patients had alimentary disease. 52 patients (28.1%) suffered from complications, which were acute respiratory distress syndrome (ARDS), secondary infection and multiple organ dysfunctional syndrome (MODS) by order. Some of the patients having complete information showed that life symptoms were stabilization. Hemogram and blood gas analysis showed that blood sugar and white blood cell (WBC) increased (P<0.001); lymphocyte decreased significantly (P<0.005) and PaO(2), PaO(2)/FiO(2) decreased Progressively (P<0.005 P<0.001) in late stage of SARS versus just entering the hospital. Half patients showed the abnormality of biochemical indexes. CONCLUSION: Age and basic diseases are sensitive indexes for predicting dead tendency of SARS. Lung injury, obstinate hypoxemia, abnormality metabolize of hyperglycemia and misusing glucocorticoids and antibiotic leading to secondary infection are the main reasons for SARS patients dying. PMID- 14552676 TI - [Evaluation of noninvasive positive pressure ventilation in treatment for patients with severe acute respiratory syndrome]. AB - OBJECTIVE: To analyze the effects of noninvasive positive pressure ventilation (NIPPV) on oxygenation of severe acute respiratory syndrome (SARS) patients, and to discuss the timing point for mechanical ventilation. METHODS: Twenty-five SARS patients with respiratory dysfunction treated with NIPPV were studied retrospectively in order to evaluate the influences within 24 hours after initiation of ventilatory support on their physiological indices and oxygenation. Patients with SARS were divided into two groups: survivor group (n=13) and non survivor group (n=12). We compared the acute physiology and chronic health evaluation (APACHEII) score, respiratory rate (RR),saturation of oxygen (SpO(2)) and modificative respiratory index (MRI) for the survivors and non-survivors before NIPPV and after NIPPV for twenty-four hours, respectively. RESULTS: Although NIPPV administered via full-face masks might be an effective treatment for rapidly improving vital signs and gas exchange and sense of dyspnea in both groups during the initial 24 hours of ventilatory support, the patients in non survivor group had higher APACHEII score, respiratory rates and lower SpO(2), MRI than the patients in survivor group (P<0.05) at the same intervals after initiation of support. CONCLUSION: Noninvasive ventilation should be used as a substitutive tool for endotracheal intubation an alternative treatment for acute respiratory failure related to SARS. Therefore, we should make efforts to avoid missing the time point for NIPPV or intubation, and we should not be restricted to the available indications for NIPPV or IPPV. PMID- 14552678 TI - [Expression of genes related to inflammatory response in myocardium of rats after burn injury]. AB - OBJECTIVE: To observe expression of genes related to inflammatory response in myocardium of rats after burn injury. METHODS: After 40 percent total body surface area (TBSA) of rat was burned gene expression of tumor necrosis factor alpha (TNF-alpha), interleukin-1beta (IL-1beta), induced nitric oxide synthase (iNOS) and cytoplastic phospholipids A(2) (cPLA(2)) in myocardium was detected with reverse transcription-polymerase chain reaction (RT-PCR) at different time after burn. Meanwhile, changes of contractile and diastolic ability of left ventricle were determined with physiological recorder. RESULTS: TNF-alpha and cPLA(2) mRNAs were increased dramatically at 1 hour after burn and maintained at high level up to 24 hours later (P<0.01), IL-1beta gene expression was up regulated at 3 hours after burn (P<0.01) and declined to normal level 12 hours later, herein any significant increase of iNOS mRNA in myocardium in rats was not found except decrease in iNOS mRNA could be detected at 1 hour after burn. As expected, severe impairment of myocardium was occurred at 3 hours after burn, represented by declining in both contractile and diasotolic ability of lefe ventrical. CONCLUSION: Over expression of TNF-alpha, cPLA(2) and IL-1beta mRNA in myocardium in rats may be partial cause of local myocardial uncontrolled inflammatory response and then partially contributed to cardiac impairment after burn. PMID- 14552679 TI - [Heat shock protein 70 induction by heat stress suppress interleukin-1beta and tumor necrosis factor-alpha levels in the brain tissue in rats with infectious brain edema]. AB - OBJECTIVE: To investigate the changes of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) in the brain tissue of rats with infectious brain edema (IBE) and their relationship with heat shock protein 70 (HSP70) by heat stress response (HSR). METHODS: Seventy-two SD rats were randomly divided into normal controls group (NS group), IBE group, and HSP group, each group was divided into three subgroups. The rats in subgroups were killed at 4 hours, 8 hours and 24 hours after the injections of IBE or normal saline respectively. HSP70 in brain tissues were determined by western blot analysis. The concentrations of IL-1beta and TNF-alpha in the brain homogenate of rats were determined by enzyme linked immunoadsorbent assay (ELISA). RESULTS: The results showed that HSP70 in brain tissues were elevated after heat shock. IBE group and NS group at 4 hours, 8 hours, 24 hours were induced to base levels of HSP70. The concentrations of TNF-alpha were significantly elevated in IBE group than in NS group at the various time points (P<0.01 or P<0.05), especially at 8 hours. The concentrations of IL-1beta were significantly increased in IBE group compared with NS group at 4 hours, 8 hours. HSR reduced the IL-1beta and TNF-alpha concentrations in the brain tissue in compared with IBE group (P<0.05 or P<0.01). CONCLUSION: IL-1beta and TNF-alpha are involved in infectious brain edema by IBE. HSP70 against infectious brain edema in rats may be associated with the reduction of IL-1beta and TNF-alpha in brain tissue. PMID- 14552680 TI - [Experimental study on recombinant human platelet-derived growth factor gel in a diabetic rat model of cutaneous incisal wound healing]. AB - OBJECTIVE: To investigate the effect of recombinant human platelet-derived growth factor (rhPDGF-BB) on cutaneous incisal wound healing in diabetic rats and this factor does-effect relationship. METHODS: Thirty Wistar diabetic rats were used in this study. Four full-thickness skin wounds of 2.54 cm(2) were incised in the back of each rat. The wounds were divided into five groups: three groups were treated with different dosage of rhPDGF-BB gel (14.0 microg/cm(2) wound, 7.0 microg/cm(2) wound, 3.5 microg/cm(2) wound), other two groups were treated with vehicle or untreated, respectively. rhPDGF-BB gel was topically applied to the wounds from 1 to 14 days. The wound healing effect was assessed by the measurements of the wound area, subcutaneous chamber and histological changes of each group. RESULTS: On 14 days, the wounds of the three groups treated with rhPDGF-BB contracted to (0.22+/-0.30)cm(2), (0.05+/-0.06)cm(2), (0.32+/ 0.32)cm(2), respectively. The area of the middle dosage groups was significantly smaller than those treated with vehicle 0.23+/-0.22 cm(2) (P<0.05) and untreated (0.22+/-0.25) cm(2) (P<0.05). On 7 days, the subcutaneous chambers of the three treated groups contracted to (0.04+/-0.03)ml, (0.02+/-0.02)ml, (0.06+/-0.03)ml, and there was a statistic significance when comparing middle dosage group with vehicle group (0.06+/-0.03) ml (P<0.05) and untreated group (0.07+/-0.05 ml (P<0.05). As for histological examination, much more granulation formation was shown in the wound bed in middle dosage group than that in other four groups on 7 days, and some of the wounds treated with middle dosage were healing on 14 days. CONCLUSION: It suggested that topical application of rhPDGF-BB might improve wound healing in diabetic rats. The effective dosage of rhPDGF-BB is 7.0 microg/cm(2) wound. PMID- 14552682 TI - [Characteristics of epidermal growth factor and its receptor expression in dermal chronic ulcers]. AB - OBJECTIVE: To investigate the expression and location of epidermal growth factor (EGF) and its receptor (EGFR) in dermal chronic ulcers and normal skin in order to explore their influence on ulcer formation. METHODS: The expression intensity and distribution of EGF and EGFR were detected with pathological method and immunohistochemistry method in 8 cases of dermal ulcers, 8 cases of edge of ulcer and 8 cases of normal skin. RESULTS: The Positive signals of EGF could be found in epidermal cells, endothelial cells and some fibroblasts; EGFR was principally located in the cytoplasm and cellular membrane of these cells mentioned above in normal skin. From normal skin, edge of ulcer to ulcerative tissues, the protein contents of EGF and EGFR were decreased progressively. In ulcerative tissues, EGF was mostly distributed in monocytes and macrophages while EGFR was chiefly sited in monocytes. When compared with normal skins, the protein expression of EGF and EGFR was notably reduced in ulcerative tissues (both P<0.01). The positive cellular ratios of two proteins were reduced to (7.1+/-5.2) % and (8.8+/-5.5) % of those in normal skin respectively (all P<0.01). CONCLUSION: The formation of dermal chronic ulcers is closely associated with the reduction of EGF and EGFR protein expression which may lead to binding obstruction between EGF and its receptor. PMID- 14552683 TI - [Effects of serum from normal and burned rats on the biological behavior of bone marrow stromal cells]. AB - OBJECTIVE: To investigate the effect of serum from normal and burned rats on the biological behaviors of bone marrow stromal cells (BMSCs). METHODS: Bone marrow was extracted from sacrificed Wistar rats and BMSCs were then incubated in F-12 medium in the presence of fetal calf serum (FCS, group F), normal rat serum (group N), serum harvested from burned rats at 3 days (group B1) and 14 days (group B2) after burn injury. The growth curve, cell cycle, apoptosis rate and contents of DNA of BMSCs were investigated. RESULTS: The growth curves of cells incubated in different rat serum were similar, but were significantly different from that of cells incubated in FCS. The doubling time of cells incubated in rat serum from group N, B1 and B2 were 23.7, 18.6 and 20.2 hours, respectively, which were significantly shortened compared with that in group F. Compared with group F, the numbers of cells at platform phase in group N, B1 and B2 were greater. The ratios of cells at phase G0/G1 in group B1 and B2 were significantly lower than those in other groups (all P<0.01), but the ratios of cells at phase S were adverse (P<0.01). Contents of DNA in cells incubated in rat serum were greater than that in cells incubated in FCS (P<0.05 or P<0.01). Contents of DNA in cells incubated in serum from burned rats were greater than that in cells in normal rat serum (P<0.01). The rate of cell apoptosis in group B1 was higher than those in other groups (P<0.01). CONCLUSION: Rat serum is more effective to stimulate proliferation of BMSCs from rat in vitro than FCS. There are probably many factors affecting the growth of BMSCs, the contents of which changed with the time after burn injury. The effects of serum from burned rats on the growth of BMSCs are complex. PMID- 14552684 TI - [Isolation and culture of multipotent mesenchymal stem cells from porcine bone marrow]. AB - OBJECTIVE: To establish a method for the isolation of porcine mesenchymal stem cells (MSCs) from bone marrow and to demonstrate their differentiation ex vivo into various mesenchymal tissue cells. METHODS: MSCs were isolated from bone marrow and purified by centrifuge and in vitro. The proliferation and growth characteristics were observed in primary and passage culture. Cell cycle was analyzed by measuring DNA content with FAC-Scan flow cytometer and cell multipotent was identified with specific staining. RESULTS: The adherent, fibroblast-like cells were confluent in single layer after plating for 12-14 days. The cultured MSCs in vitro differentiated into osteoblasts. The cell cycle analysis showed that 80% of MSCs were in G0/G1 phase. CONCLUSION: Porcine MSCs can be isolated from postnatal bone marrow through their adherent ability. Porcine MSCs may be introduced as a valuable model system to study the mesenchymal lineages for basic research and tissue engineering. PMID- 14552685 TI - [Changes and effects of group II metabotropic glutamate receptors in the cerebral cortex of rats after diffuse brain injuries coupled with hypopiesia secondary brain insults]. AB - OBJECTIVE: To study the changes and effects of group II metabotropic glutamate receptors (mGluR) after diffuse brain injuries (DBI) coupled with hypopiesia secondary brain insults (SBI). METHODS: Male SD rats were randomized into four groups: normal control, sham-operated, DBI alone and DBI coupled with SBI group. The SBI model was made on the basis of Marmarou's model. The mRNAs of the mGluRs were detected at 1, 3, 6, 12, 24, 48 and 72 hours after injuries by in-situ hybridization, and the positive neurons were counted. RESULTS: Statistical analysis showed no significance between normal control group and sham-operated group, DBI group in mGluR 2, 3 mRNA (all P>0.05). The number of mGluR 2, 3 positive neurons decreased at 12 hours after injury and the peak occurred at 48 hours after injury in the injured cerebral cortex in DBI alone group (both P<0.05). However, in DBI with SBI group, there was a significant decrease of the number of mGluR2, 3 positive neurons at 6 hours after injury and the peak happened at 24 hours after injury (both P<0.05). CONCLUSION: mGluR2, 3, factors with the function of protecting brain, might play an important role in the path physiology of DBI and SBI. PMID- 14552686 TI - [Changes in proinflammatory cytokines and immune function of subeschar tissue fluid during early stage after severe burn]. AB - OBJECTIVE: To evaluate the inflammatory response and immune function of subeschar tissue fluid (STF) in early stage of severe burn. METHODS: Eight patients were observed and samples from blood as well as STF were collected at 16, 24, 32, 40, 48 hours following burn to measure white blood cell (WBC) counts, levels of immunoglobin (IgA, IgG, IgM), complement (C3, C4) values as well as proinflammatory cytokines as tumor necrosis factor-alpha (TNF-alpha), interleukin 6(IL-6) and IL-8. In addition, twenty Balb-c mice were used for the lethal experiment by STF collected at 48 hours after burn. RESULTS: The levels of IgA, IgG, IgM in STF were about half of those in the blood samples (all P<0.01). The WBC counts in STF were 1/100 time of that in the blood (all P<0.01). The contents of TNF-alpha, IL-6 and IL-8 in STF were comparable with those in the blood (P>0.05). Following STF challenge, none of the mice died during 7-day observation period. CONCLUSION: The cells and large molecules seems to be more difficult to enter STF compared with small molecules. No marked local inflammatory response occurs in STF during early stage of severe burn, and STF challenge has no lethal effect. The low WBC counts and low immunoglobin as well as complement levels imply the limited effect of STF on local infection, and effective antibiotics should be timely used after major burn. PMID- 14552687 TI - [Dynamic changes of platelet number in early multiple injury and it is effect on prognosis]. AB - OBJECTIVE: To study the dynamic changes of platelet number in early multiple injured patients and its clinical significance. METHODS: Ninety patients with multiple injury less than 48 hours were selected. Blood analysis was dynamicly investigated at 1, 24, 72, 120 and 168 hours after in Hospital, and acute physiology and chronic health evaluation III (APACHE III) and injury seriousness scale (ISS) scores were recorded. RESULTS: The platelet number declined after injury, reached the lowest point at 72 hours after in hospital, then restored to the normal level at 120 hours. In the platelet decrease group, the trauma was more critically[ISS (29.60+/-9.80) scores vs. (22.30+/-12.00) scores, (P<0.05)] and the incidence of multiple organs dysfunction syndrome (MODS) was higher than normal group (83.9% vs. 62.7%, P<0.05). In the death group, the number of platelet was declined[ (30.1+/-10.3) hours vs. (51.3+/-14.8) hours, (P<0.05) and the lasting tine was longer[ (63.7+/-11.0) hours vs. (54.2+/-13.4) hours, (P<0.05). CONCLUSION: The platelet number declines after multiple injury. The decrease and continuously lower level of platelet suggest poor prognosis. PMID- 14552688 TI - [Change in intestinal free radical and prostaglandin metabolism in the aged rats with pneumococcus pneumonia]. AB - OBJECTIVE: To study the pathogenic mechanism underlying intestinal injury in the aged rat with pneumonia. METHODS: The model of rats with pneumococcus pneumonia was reproduced, and animals were divided into young control group (YCG), young model group (YMG), aged control group (ACG) and aged model group (AMG). The pathological change of lung tissue and intestine, contents of intestinal 6-keto prostaglandin F(1alpha) (6-keto-PGF(1alpha)), thromboxane B(2) (TXB(2)), nitric oxide (NO), malondialdehyde (MDA) and superoxide dismutase (SOD) activity were determined in various groups. RESULTS: The lung and intestinal injury in AMG was more serious than that in the YMG. The decreases in SOD activity and 6-keto PGF(1alpha) content, the increases in contents of TXB(2), NO, MDA in the YMG and the AMG were obvious than those in the YCG and the ACG respectively. The decrease in SOD activity and increase in MDA content were obvious in the ACG than those in the YCG. In addition, the decrease in SOD activity and increase in MDA content in the AMG were significant compared with those in the YMG. CONCLUSION: The Prostaglandin metabolism and the free radical injury might be in the pathogenesis of intestinal injury in the aged rats with pneumonia. The intestinal injury induced by free radical in aged rats with pneumonia appears to be more obvious with ageing. PMID- 14552689 TI - [Effect of endotoxemia on thyrotropin releasing hormone and its receptor in Wistar rat brain]. AB - OBJECTIVE: To study the dose and location of thyrotropin releasing hormone (TRH) and its receptor in Wistar rat's brain when early endotoxemia happened. METHODS: Twenty-two Wistar rats were selected. In 13 mouse the model was established with endotoxemia and hemorrhagic shock. The TRH and its receptor were observed with radioimmunoassay (RIA), and were compared to the 9 control rats. RESULTS: The dose of TRH decreased significantly in the early stage of endotoxemia. The maxium dose (Bmax) of TRH increased while Kd (affinity) decreased, so the effective TRH receptor had no significantly change. CONCLUSION: The dose of TRH is different in brain and serum. The decrease of TRH and the steady of active TRH receptor may protect the brain tissue and repress multiple organ dysfunction syndrome (MODS) in the early stage of endotoxemia. PMID- 14552690 TI - [Change of the erythrocyte chemokine receptor binding activity in scalded rats with Pseudomonas aeruginosa infection]. AB - OBJECTIVE: To explore the influence of infection on the erythrocyte chemokine receptor (ECKR) binding activity in severely scalded rats. METHODS: The Sprague Dawley (SD) rats were randomly divided into three group: sham scald group (A), burn and infection group (B) and infection group (C). The B group rats were scalded with 30% total body surface area (TBSA) of III degree, and the rats ECKR binding activity with interleukin-8 (IL-8) as ligand were detected by enzyme linked immunosorbent assay (ELISA) at 1, 2, 4, 6 and 8 hours after infection. RESULTS: Compared to that in A group, ECKR binding activities declined significantly (both P<0.01) after infection in both of B and C groups, but they increased at 8 hours (P<0.01). ECKR binding activity in B group was significantly less at 2, 8 hours than that in C group after infection (both P<0.05). The declining range of ECKR binding activity was more in B group, in which the decline of ECKR binding activity appeared earlier (2 hours) than that in C (6 hours) group. CONCLUSION: The infection lead to the decline of ECKR binding activity in burned rats, and the erythrocytes might participate in the chemokine regulation and play a novel role in the infection. PMID- 14552691 TI - [Effect of hypotonic liquid containing colloid on hemorrhagic dogs with celiac seawater immersion wound]. AB - OBJECTIVE: To determine hypotonic liquid containing colloid on hemorrhagic dogs with celiac seawater immersion wound. METHODS: Thirty-five dogs were randomly divided into five groups: control group (group A), 0.9% NaCl therapy group (group B) and 0.45% NaCl therapy group (group C), 5% glucose therapy group(group D) and hypotonic liquid containing colloid therapy group (group E). Changes of mean artery pressure (MAP), cardiac output (CO), urine volume, plasma osmotic pressure and pathologic changes of lung and brain were observed. RESULTS: hypotonic liquid containing colloid could effectively ameliorate MAP and CO, improve urine volume, decrease plasma osmotic pressure and prevent occurrence of lung and brain edema. CONCLUSION: Hypotonic liquid containing colloid can exert satisfactory therapeutic effects on hemorrhagic dogs with celiac seawater immersion wound. PMID- 14552692 TI - [Risk factors for intracerebral hemorrhage after intravenous thrombolysis in acute cerebral infarction]. AB - OBJECTIVE: To identify risk factors of intracerebral hemorrhage (ICH) after intravenous thrombolysis in acute cerebral infarction. METHODS: Eighty-two patients with acute cerebral infarction were treated with intravenous urokinase within 12 hours from stroke onset. The data of clinic and laboratory were as readily available baseline factors and were analyzed to select potential contributors to ICH. RESULTS: ICH occurred in 15 patients (18.3%), including 6 hemorrhagic infarction (HI, 7.3%) and 9 Parenchyma hemorrhage (PH, 11.0%). Seven patients had symptomatic ICH, and 4 patients died. Patients with ICH had significantly lower European stroke scale (ESS) scores. Patients with ESS<60 scores, early ischemic changes (EIC) on cranial CT scans or artrial fibrillation had significantly higher risk of ICH. CONCLUSION: Risk factors associated with ICH are severity of neurological deficit, EIC and artrial fibrillation. PMID- 14552697 TI - COX-2 and COX-2 enzyme inhibition. PMID- 14552698 TI - The prospective use of COX-2 inhibitors for the treatment of temporomandibular joint inflammatory disorders. AB - Development of a new class of drugs designed to selectively inhibit the inducible cyclooxygenase isoenzyme, COX-2, was initially prescribed for individuals diagnosed with osteoarthritis or rheumatoid arthritis. Although these inflammatory disorders are more typically related to the joints of the knee, ankle, or hand, the temporomandibular joint (TMJ) plays a special role due to its involvement in our normal day-to-day activities of eating and communicating. The TMJ, unlike most of the other joints, contains some unique morphological characteristics that support various inflammatory disorders. An overview of these characteristics and the prospective use of the COX-2 inhibitors for temporomandibular joint inflammation are presented. PMID- 14552699 TI - COX-2 specific inhibitors in NSAID-intolerant patients. AB - Most adverse NSAID-induced respiratory and skin reactions appear to be precipitated by the inhibition of cyclooxygenase-1 (COX-1); this in turn activates the lypoxygenase pathway, which eventually increases the release of cysteinyl leukotrienes (Cys-LTs). Recent studies have reported that patients with NSAID-induced asthma have a low production of PGE2 in respiratory epithelial cells, bronchial fibroblast and peripheral blood cells. Low production of PGE2 may be due to an insufficient cyclooxygenase-2 (COX-2) expression in the inflammatory response underlying asthma. Since PGE2 administered by inhalation inhibits NSAID-induced bronchoconstriction and the parallel increase in Cys-LTs release, a reduced PGE2 synthesis may render NSAID-patients more susceptible to the COX-1 inhibitory effects of NSAIDs. Recent studies have shown that selective COX-2 inhibitors (rofecoxib and celecoxib), unlike COX-1 inhibitors, are very well tolerated by NSAID-sensitive patients and do not elicit increased Cyst-LTs production. However, these drugs can still can precipitate cutaneous reactions in a significant proportion of patients with skin reactions to NSAID. The heterogeneity of the NSAID-intolerance syndrome suggests that subjects who do not tolerate NSAID can use coxibs only after first having been exposed to the drug under the supervision of a specialist with experience in these procedures. PMID- 14552700 TI - Efficacy, safety and potential clinical roles of the COX-2-specific inhibitors. AB - About three decades ago, the primary mechanism of action of the non-steroidal anti-inflammatory drugs (NSAIDs) was elucidated when their inhibitory action on the production of prostaglandins was discovered. It was found that their action is centred on the inhibition of the cyclooxygenase (COX) enzyme responsible for converting arachidonic acid to prostaglandins. More recently, it was discovered that at least two isoforms of the enzyme exist. While the COX-1 isoenzyme is mainly constitutive, the COX-2 form is inducible. The constitutive form is principally associated with physiological/homeostatic functions while the inducible form is involved in inflammatory responses. Because many of the unwanted effects of the NSAIDs have been associated with COX-1 inhibition, specific drugs with a high inhibitory activity toward the COX-2 isoenzyme have been developed in recent years. The efficacy, safety and potential clinical roles of these COX-2-specific inhibitors are addressed in this review article. PMID- 14552701 TI - COX-2-selective inhibitors (COXIBs): gastrointestinal safety. AB - COX-2 selective inhibitors (coxibs) have been developed with the primary aim to reduce/avoid gastrointestinal (GI) toxicity observed during conventional (non selective) non-steroidal anti-inflammatory (NSAID) therapy. Coxibs have clearly and convincingly been shown to be superior to conventional NSAIDs with significantly less GI side effects. When hard endpoints such as perforation, obstruction, and serious bleeding considered, coxibs reduce the risk by approximately 50 percent. Although selective COX-2 inhibition seems not to be enough for complete elimination of GI toxicity, coxibs posses no more GI toxicity than placebo in prospective clinical studies and further increase in COX-2 selectivity does not reduce GI toxicity. For the initial aim developed, thus coxibs fulfilled their promise and will soon replace conventional NSAIDs. PMID- 14552702 TI - Experimental brain inflammation and neurodegeneration as model of Alzheimer's disease: protective effects of selective COX-2 inhibitors. AB - Epidemiological studies indicate that long-term treatment with non-steroidal anti inflammatory drugs reduces the risk of Alzheimer Disease and may delay its onset or slow its progression. Neuroinflammation occurs in vulnerable regions of the Alzheimer's disease (AD) brain where highly insoluble beta-amyloid (Abeta) peptide deposits and neurofibrillary tangles, as well as damaged neurons and neurites, provide stimuli for inflammation. To elucidate the complex role of inflammation in neurodegenerative processes and the efficacy of selective COX-2 inhibitors in AD, we examined whether the attenuation of brain inflammatory reaction by selective COX-2 inhibitors may protect neurons against neurodegeneration. The data reported in this review show that in in vivo models of brain inflammation and neurodegeneration, the administration of selective COX 2 inhibitors prevent not only the inflammatory reaction, but also the cholinergic hypofunction. Our data may help elucidate the epidemiological findings indicating that anti-inflammatory agents, in particular NSAIDs, reduce the risk of developing AD and may slow its progression. PMID- 14552703 TI - Immune restoration in head and neck cancer patients via cyclooxygenase inhibition: an update. AB - Most carcinomas overexpress cyclooxygenase, especially COX-2, thus secreting large amounts of immunosuppressive prostaglandins. Epidemiological data and animal models have provided evidence that inhibition of cyclooxygenase and thus prostaglandin E2 synthesis via non-steroid antiinflammatory drugs (NSAIDs) inhibits tumor growth in vitro and in vivo. Moreover, it could be demonstrated that chemoprevention, i.e. the long-term use of NSAIDs, significantly reduced the risk of developing certain types of cancer. However, the molecular mechanisms underlying these antineoplastic effects are not entirely understood. This review focuses on prostaglandin-mediated immunosuppressive mechanisms in head and neck cancer and presents immunorestorative strategies via cyclooxygenase inhibition in vitro and in vivo with special emphasis on COX-2. A better understanding of the interaction of tumors with the immune system and how the process of carcinogenesis can be antagonized by selectively modulating the activity of specific enzymes such as COX-2 will provide the rationale for the use of NSAIDs for chemoprevention or immunoadjuvant cancer therapies. PMID- 14552704 TI - Clinical pharmacology of selective COX-2 inhibitors. AB - The discovery of cyclooxygenase (COX)-2 has provided the rationale for the development of a new class of nonsteroidal antiinflammatory drugs (NSAIDs), the selective COX-2 inhibitors (denominated coxibs), with the aim of reducing the gastrointestinal (GI) toxicity associated with the administration of NSAIDs by virtue of COX-1 sparing. Rofecoxib and celecoxib are the first selective COX-2 inhibitors approved by the FDA and EMEA for the treatment of rheumatoid arthritis (RA), osteoarthritis (OA) and for relief of acute pain. Rofecoxib has been shown to spare COX-1 activity ex vivo, in platelets and gastric mucosa, when administered at therapeutic doses or above. In a large clinical trial, COX-2 inhibitors have been demonstrated to halve the incidence of serious upper GI events vs a nonselective NSAID. Recently, other selective COX-2 inhibitors with different COX-1/COX-2 selectivity and pharmacokinetic features have been developed, i.e. valdecoxib, parecoxib, etoricoxib and lumiracoxib. The improved biochemical selectivity of valdecoxib vs celecoxib in vitro (COX-1/COX-2 ratio: 60 vs 30, respectively) may be clinically relevant leading to an improved GI safety. Interestingly, parecoxib, a pro-drug of valdecoxib, is the only injectable coxib. Etoricoxib, showing only a slightly higher COX-2 selectivity than rofecoxib in vitro (COX-1/COX-2 ratio: 344 vs 272, respectively), has been reported to cause a similar specific COX-2 inhibition ex vivo that should translate into comparable GI safety. Lumiracoxib, the most selective COX-2 inhibitor in vitro (COX-1/COX-2 ratio: 400), is the only acidic coxib. It has been hypothesized that this pecular chemical feature may lead to an enhanced concentration in inflammatory sites that may translate into an improved clinical efficacy. The results of clinical trials have shown that coxibs have a comparable clinical efficacy and renal toxicity and an improved GI safety vs nonselective NSAIDs. Whether the different pharmacodynamic and pharmacokinetics features of the various coxibs will produce detectable differences in efficacy and toxicity remains to be evaluated in appropriate comparative randomized clinical studies. PMID- 14552705 TI - Transcriptional regulation of cyclo-oxygenase expression: three pillars of control. AB - Blocking cyclo-oxygenase (COX) isoform activities with non-steroidal anti inflammatory drugs (NSAIDS) is widely employed in the treatment of arthritis. These agents also hold great promise in the context of pre and post-neoplastic diseases such as colorectal cancer (CRC). Nevertheless, issues of isoform specificity and delivery necessitate the exploration of other strategies to specifically block expression of the COX genes. Approaches that target gene transcription may complement enzyme inhibition. Thus, understanding the regulation of COX isoform transcription may improve the specific inhibition of expression. Three tiers of transcriptional regulation are evident: initiation, alternative splicing and messenger RNA stability. Transcription factors that activate COX-2 expression are elevated in certain disease states and emergency responses such as infection and are therefore potential targets. These factors include C/EBP-beta, phospho- CREB, NF-IL6, AP1, NFkB, and TCF-4/LEF-1. In this review we highlight another factor, c-MYB as a key COX-2 regulator in CRC. Alternative exon usage is another tier of regulation that has not received much attention. For instance, COX-1 splice variants (also known as COX-3 and PCOX-1a) may broaden the spectrum of COX activities in disease. Similarly, whilst mRNA stability is clearly modulated by steroids in the case of COX-2, the wider implications of targeting mRNA stability have not been afforded the same attention. Finally, it seems that some NSAIDS exert part of their action directly on COX-2 transcriptional regulation explaining why such agents display greater effects on this isoform than enzyme inhibition data would suggest. PMID- 14552706 TI - [Medical imaging investigation and project of hepatocellular carcinoma]. PMID- 14552707 TI - [Basic and applied studies on the vaccines against hepatitis B virus]. PMID- 14552708 TI - [Viral hepatitis combined with malignant histiocytosis: a case report]. PMID- 14552709 TI - [Pilot study on clinical application of hepatic perfusion with multi-slice spiral CT]. AB - OBJECTIVE: To research the value of hepatic perfusion with multi-slice spiral CT in the diagnosis of liver diseases. METHODS: Among the 48 patients undergone dynamic CT of the liver, 20 were volunteers without hepatic disorder, 17 with cirrhosis, 11 suffered from hepatic cancer. The perfusion indexes were calculated and compared. RESULTS: (1) Compared with the control group, HPP (ml/min/ml), PPI and HPP/HAP of patients with cirrhosis were significant lower (HPP: 0.49+/-0.19 vs 0.60+/-0.16, P=0.038; PPI: 0.58+/-0.14 vs 0.67+/-0.06, P=0.015; HPP/HAP: 1.63+/-0.87 vs 2.12+/-0.65, P=0.04), whereas HPI was higher (0.42+/-0.14 vs 0.33+/-0.06, P= 0.015), which indicated the decrease of portal inflow and the increase of arterial inflow in cirrhosis patients. (2) Patients with hepatic cancer got a significant higher average HAP than that in volunteers and cirrhosis patients (F=11.71, P<0.001), while their HPP and HPP/HAP showed significant declining (F=22.84, P=0.0001; F=20.67, P<0.0001, respectively), which implied that hepatic cancer was mainly supplied by artery. CONCLUSIONS: Hepatic perfusion with multi-slice spiral CT is an non-invasive technique to evaluate the arterial and portal inflow separately, which can inflect the hemodynamic change of the lesion by the perfusion indexes, and identify the condition of the tissue round the lesion prior to morphologic change. This method shows important value of diagnosis and differential diagnosis in hepatic diseases. PMID- 14552710 TI - [Clinical value of MR cholangiopancreatography combining with dynamic enhanced MRI in the detection of cholangiocarcinoma]. AB - OBJECTIVES: To evaluate the clinical value of breath-hold magnetic resonance cholangiopancreatography (MRCP) combining with dynamic enhanced MRI in the diagnosis of cholangiocarcinoma. METHODS: MRCP findings of 88 cholangiocarcinoma patients proved surgically and pathologically were analyzed retrospectively. RESULTS: MRCP examination succeeded in all the 88 patients and the pancreaticobiliary ducts were shown satisfactorily. The accuracy of MRCP in the location of both hilar and extrahepatic cholangiocarcinoma was 100%, and the accuracy of detecting hilar and extrahepatic cholangiocarcinoma were 100% and 52.2%, respectively. Combining with dynamic enhanced MRI, the detecting accuracy of extrahepatic cholangiocarcinoma improved to 91.3%. CONCLUSIONS: MRCP examination has a high successful rate and can accurately determine the location of hilar and extrahepatic cholangiocarcinoma, and the accuracy of qualitative diagnosis for the former two is high. Combining with dynamic enhanced MRI, the specificity of determining extrahepatic cholangiocarcinoma is also high. PMID- 14552711 TI - [Small hepatocellular carcinoma: multiphase hepatic CT scan with a multidetector row helical CT scanner]. AB - OBJECTIVES: To evaluate multiphase hepatic CT scan using multidetector row helical CT (MDCT) in detecting hypervascular small hepatocellular carcinoma (SHCC). METHODS: Multiphase hepatic CT scan in 75 patients with SHCC was carried out with Marconi MX8000 multidetector row helical CT scanner. The early arterial phase scan started at the 21th second after the injection of contrast medium, the late arterial phase scan started at the 34th second, and the portal venous phase started at the 80tieth second, six seconds every time. RESULTS: The study showed that the density values of 71 lesions between liver and SHCC were different significantly among the early arterial phase, the late arterial phase and the portal venous phase (F=3.327, P<0.05). Among the 91 lesions, the detectable rates of the early arterial phase and the late arterial phase were 45.1% and 83.5%, respectively. The rate of double arterial phases was 92.3%. The rates increased to 94.5% and 97.8%, when the late arterial phase combined with the portal venous phase and the double arterial phase combined with the portal venous phase. CONCLUSION: The utility of MDCT with faster speed, thinner slice and multiphases scanning has improved the opportunity of detecting hypervascular hepatocellular carcinoma. PMID- 14552712 TI - [Relationship between the HBV core gene mutation and the cellular immunity in host]. AB - OBJECTIVES: To study the relationship between the mutation of Leu60Val in HBV core region and the cellular immunity in patients with chronic hepatitis B (CHB). METHODS: HBV DNA C gene mutation was confirmed by polymerase chain reaction (PCR) and sequencing the products directly. The cytokines (IFN-gamma, TNF-alpha and IL 2) levels in serum were measured by enzyme linked immunosorbent assay (ELISA). The distribution of T-lymphocyte subpopulations in peripheral blood was detected by flow cytometry (FCM). RESULTS: The mutation of Leu60Val was found in 19 out of the 91 CHB patients. With the CHB severity, the mutation rate was getting higher, especially in the severe hepatitis group. The IFN-gamma and TNF-alpha levels were much higher in mutant strain group than those in wild strain group (t=2.584, 4.766, P<0.01), so was the ratio of CD4+/CD8+ (t=2.275, P<0.05). CONCLUSION: The mutant strain of 60Val may increase affinity to HLA-I molecule, or up-regulate the expression of HLA-I molecule, resulting in the activation of CTL to release the cytokines and cause immune response in liver. PMID- 14552713 TI - [The role of cyclooxygenase 2 and prostaglandin I2 in the development of portal hypertensive gastropathy]. AB - OBJECTIVES: To study the role of cyclooxygenase 2 (COX 2) and prostaglandin I2 (PGI2) in the development of portal hypertensive gastropathy (PHG). METHODS: Forty Wistar rats were divided into surgery group (32) and control group (8). Partial portal vein ligation method was used to narrow the sectional area of portal vein to establish the experimental model of PHG in surgery group rats. Then they were divided into four groups (8 rats in each). The free pressure of portal vein was determined at the 1st, 2nd, 3rd, 4th weeks after the operation, and 8 rats were killed to observe the pathological change of gastric mucosa. The levels of 6-keto-PGF1 alpha, a stable metabolite of PGI2, were determined by radioimmunoassay in gastric mucosa homogenate and the blood of portal vein. The expression of COX 2 in gastric mucosa was determined by immunohistochemistry. RESULTS: The free pressure of portal vein increased rapidly after partial portal vein ligation and maintained a high stable level after 1 week. They were (2.40+/ 0.15) kPa, (2.38+/-0.17) kPa, (2.52+/-0.21) kPa, and (2.46+/-0.17) kPa at the 1st, 2nd, 3rd, and 4th weeks after partial portal vein ligation, while it was (0.90+/-0.16) kPa in control group (t>or=17.356, P<0.05). The gastric mucosa appeared pale, edema, hyperaemia, surface erosion, punctate hemorrhage and these lesions were more apparent with the time after the operation. The pathological examination showed that the gastric mucosa and submucosa thickened. The vessels of gastric mucosa and submucosa expanded and increased. There were lymphocytes and neutrophils infiltration around the vessels in the gastric mucosa and submucosa. The 6-keto-PGF1 alpha levels in gastric mucosa and the blood of portal vein increased rapidly and maintained a high level after partial portal vein ligation,which were higher than those in control group (104.52pg/ml+/-25.11pg/ml vs 73.62pg/ml+/- 20.33pg/ml, t=2.710, P<0.05; 180.21pg /ml+/-37.56pg /ml vs 142.11pg /ml+/-31.51pg /ml, t=2.198, P<0.05). The results of immunohistochemistry showed that the intensity and degree of the COX 2 staining in gastric tissue increased at the 1st, 2nd, 3rd, 4th weeks after partial portal vein ligation, while the COX 2 in control group rats was negative. CONCLUSIONS: The expression of COX 2 and PGI2 in gastric tissue increased in portal hypertension. PGI2 as an inflammatory medium, damages the gastric mucosa by expanding vessels and other mechanisms in portal hypertension. It may be one of the important factors contributing to the development of PHG. PMID- 14552714 TI - [Effects of cisapride on intestinal bacterial and endotoxin translocation in cirrhotic rats]. AB - OBJECTIVES: To further investigate the effects of cisapride on intestinal bacterial overgrowth (IBO), bacterial and endotoxin translocation, intestinal transit and permeability in cirrhotic rats. METHODS: 25 normal control rats, 25 cirrhotic rats, 20 cirrhotic rats received saline, and 20 cirrhotic rats treated with cisapride were included in the study. All animals were assessed with many variables including bacterial and endotoxin translocation, IBO, intestinal transit and permeability. RESULTS: Bacterial translocation was found in 48%(12/25) cirrhotic rats and none of control rats. Among the 20 rats with IBO, there were 11 rats with bacterial translocation (BT) while only one rats occurred BT out of the 5 rats without IBO. Cirrhotic rats with IBO had a significantly higher rate of endotoxin translocation, higher intestinal permeability and longer intestinal transit than those without IBO. BT of a specific organism was always associated with IBO of that organism. Compared with the placebo group, cisapride treated rats had lower rates of bacterial and endotoxin translocation and IBO, which had close relationship with shorter intestinal transit and lower permeability. CONCLUSION: Endotoxin and bacterial translocation in cirrhotic rats may be the result of IBO and higher permeability. IBO may be the result of longer transit. Cisapride which can accelerate intestinal transit and improve intestinal permeability is helpful in preventing and treating intestinal bacterial and endotoxin translocation. PMID- 14552715 TI - [Inhibitive effects of recombinant adenovirus-mediated human endostatin on the growth of human hepatocellular carcinoma xenograft in nude mice]. AB - OBJECTIVE: To investigate the inhibitive effect on the growth of hepatocellular carcinoma (HCC) xenografted in nude mice by adenovirus-mediated human endostatin gene. METHODS: The expression efficiency of endostatin was examined after ECV-304 cells infected with Ad/hEndo by western blot. The hepatoma BEL-7402 cells were injected into Balb/c nude mice to detect the inhibition of Ad/hEndo on the growth of HCC xenografted in nude mice. The expression of endostatin mRNA in tumor tissue was analyzed with RT-PCR, and its distribution in vivo was also analyzed. RESULTS: High level expression of endostatin achieved in infected ECV-304 cells by western blot. Ad/hEndo significantly inhibited the growth of xenografted BEL 7402 tumors (F=4.061, P<0.05). The intratumoral microvessel density (MVD) decreased significantly in the treated mice (6.88+/-1.08 vs 13.60+/-1.71, t=9.216, P<0.01). The expression of endostatin mRNA in tumor tissue was detected by RT-PCR in 3 days after administration intratumorally with Ad/hEndo and almost disappeared in 7 days. Endostatin mRNA was mainly located in tumor tissue with a higher concentration than that in heart, lung, spleen and liver after Ad/hEndo administration. CONCLUSION: Adenovirus-mediated human endostatin gene can be expressed efficiently in vitro and in vivo, and significantly inhibit the growth of BEL-7402 xenografted tumors in nude mice. PMID- 14552716 TI - [Apoptosis in hepatoma cells induced by antisense oligodeoxynucleotide against survivin]. AB - OBJECTIVES: To investigate the apoptosis induced by antisense oligodeoxynucleotide (ASODN) against survivin and the mechanisms after the hepatocellular carcinoma SMMC-7721 cells transfected with the ASODN. METHODS: The ASODN was transfected into SMMC-7721 cells mediated by liposomal reagent. The changes of cell cycle and apoptotic rate were detected by flow cytometry. The changes of cell skeleton was observed through confocal microscope. The activity of p38MAPK and caspase-3 were detected by immuno-precipitation and kinase activity assess methods, respectively. RESULTS: There were control, sense control, 400, 600, 800, and 1 000 ng/ml ASODN groups (I - VI). The apoptotic rats were 0.70%, 0.76%, 2.43%, 7.82%, 23.11%, and 31.35% in groups I - VI, respectively, which in the ASODN-transfected groups were higher than that in the control group (tor=20.9, P<0.01). The activity of p38MAPK increased significantly, when the ASODN was transfected at the concentration of 600 ng/ml or more, so did the caspase-3 activity (the p38MAPK and caspase-3 activity in groups I - VI were 7.03, 7.07, 13.47, 16.37, 43.97, 47.87 and 0.015+/-0.010, 0.014+/-0.002, 0.026+/-0.003, 0.042+/-0.001, 0.093+/ 0.001, 0.100+/-0.001, respectively). CONCLUSIONS: ASODN targeting at survivin mRNA can induce G2/M stop, activate p38MAPK and caspase-3. The activated caspase 3 destroys the cell skeleton microfilament system, resulting in apoptosis. PMID- 14552717 TI - [A modified method detecting apoptotic cells using Apo2.7 monoclonal antibody]. PMID- 14552718 TI - [Relationship between aberrant FHIT transcripts and hepatocellular carcinoma]. AB - OBJECTIVE: To study the relationship between aberrant FHIT transcripts and hepatocellular carcinoma (HCC). METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) and single strand conformational polymorphism (SSCP) assays were used to analyze the transcripts and mutations of FHIT gene in 24 matched tumorous tissues and para-tumorous tissues from patients with HCC and in 4 normal liver tissues. RESULTS: Aberrant FHIT transcripts were observed in 11 out of 24 (46%) tumorous tissues and in 2 (8%) of the matched para-tumorous tissues. CONCLUSION: FHIT aberrant transcripts may play an important role in the pathogenesis of hepatocellular carcinoma. PMID- 14552719 TI - [Relationship between the expression of IL-10 and liver regeneration following reduced-size orthotopic liver transplantation in rats]. AB - OBJECTIVES: To explore the relationship between the expression of IL-10 and liver regeneration following reduced-size orthotopic liver transplantation in rats. METHODS: Rats models with reduced-size orthotopic liver transplantation were established. The rats were divided in three groups: partial liver resection (I), orthotopic liver transplantations (II), and reduced-size orthotopic liver transplantation (III). The expression of IL-10 and regenerative response of liver in rats were evaluated by immunohistochemistry and flow cytometry on the 1st, 2nd, 4th and 7th days after the operations, respectively. RESULTS: The liver grafts were capable of regeneration, the proliferation activity peaked on the fourth day with 26.3+/-0.9, 35.8+/-2.2, and 32.4+/-1.8 in I, II, and III groups, respectively. The expression of IL-10 was negative correlation to liver regeneration (r=-0.58, P<0.01). CONCLUSIONS: Whole and reduced-size transplanted livers show the same regenerative activity. The maximal regenerative response delayes slightly, compared with that after partial hepatectomy. IL-10 plays an important immunomodulatory role in liver regeneration,and the effect is affected by general immune system and other cytokines. PMID- 14552720 TI - [Liver histological changes after lamivudine treating in chronic hepatitis B patients with HBeAg positive]. AB - OBJECTIVES: To investigate the histological changes in liver biopsy tissues taken from chronic hepatitis B patients with HBsAg and HBeAg positive and ALT abnormal after lamivudine therapy for one year. METHODS: Lamivudine was given orally at the dose of 100 mg once a day for one year. 101 patients were enrolled into this open-label study. Paired liver biopsies from patients with hepatitis B before and after therapy with lamivudine were studied. Blinded biopsies were evaluated by a histopathologist and scored according to Knodell's histology activity index(HAI). RESULTS: 53.5% (54/101), 51.5% (52/101) and 31.7% (32/101) patients had a reduction of their total hepatic HAI score, necroinflammation and fibrosis scores by >or=2 points or 1 points at the end of one year of lamivudine therapy, compared with their pretreatment values, respectively. There were significant reduction of HAI score, necroinflammation and fibrosis scores from 8.0+/-4.7 to 5.2+/-3.3 (t=7.358, P<0.01), from 5.9+/-3.8 to 3.6+/-2.5 (t=7.298, P<0.01), and from 2.1+/-1.2 to 1.6+/-1.2 (t=3.827, P<0.01), respectively. The histological improvement was independent on the HBeAg seroconvertion during the therapy. CONCLUSION: Significant improvement in liver histology, both necroinflammation and fibrosis, can be obtained in the majority of patients treated with lamivudine for one year. PMID- 14552721 TI - [Clinical analysis of 19 patients with acute toxic hepatitis by 1,1,1 trichloroethane]. PMID- 14552722 TI - [Individual therapy for the patients with YMDD mutation in HBV by lamivudine]. PMID- 14552724 TI - [An overview of diagnostic imaging for liver cancer]. PMID- 14552723 TI - [Ten years outcome observation of infants with hepatitis C virus mother to infant transmission]. AB - OBJECTIVE: To study the outcome of hepatitis C virus (HCV) vertical transmitted infants. METHODS: Thirteen HCV vertical infected infants were followed up for 10 years. HCV antibody and HCV RNA in the blood samples from them were tested using second generation HCV antibody EIA kits and RT-PCR, respectively. RESULTS: Among the 13 infants, one developed clinical hepatitis C, and serum HCV antibody and HCV RNA could be detected for 7 and 8 years, respectively. Three were subclinical hepatitis C, serum HCV antibody continued to be positive for 12 months (2 infants) and 24 months (1 infant), respectively, and serum HCV RNA turned to be negative at the 24th month (2 infants) and the 60th month (1 infant), respectively. Nine were HCV insidious infection, whose serum HCV antibody and HCV RNA turned to be negative in 12 months. During the eight to ten years, there was no infants with anti-HCV or HCV RNA positive again. CONCLUSIONS: It is rarely happened that vertical transmitted HCV induce chronic HCV carrying state and chronic viral hepatitis, and most of the infected infants have good outcome. PMID- 14552725 TI - [Imaging detecting technologies for liver cancer and selection]. PMID- 14552726 TI - [The value of CT and MRI in the characterization for hepatocellular carcinoma]. PMID- 14552727 TI - [Imagings of helical CT scan and changes of blood supply in small hepatocellular carcinoma]. PMID- 14552728 TI - [Magnetic resonance imaging evaluating the effects of percutaneous radiofrequency ablation on liver malignancies]. PMID- 14552729 TI - [A color Doppler ultrasonographic study on patients with middle and late-stage liver cancer treated by transcatheter arterial chemoembolization]. PMID- 14552730 TI - [Treatment effect of transcatheter hepatic arterial chemoembolization combining with IL-2 on primary hepatoma]. PMID- 14552731 TI - [Application of time-resolved fluoroimmunoassay in the detection of HBV markers]. PMID- 14552732 TI - [Effect of batimastat on the dynamic changes of matrix metalloproteinases in liver tissue during the development of hepatocellular carcinoma in rats]. PMID- 14552733 TI - [Primary study on the effect of different dimethyl sulfoxide concentrations on 80 degrees C cryopreserved porcine hepatocytes]. PMID- 14552734 TI - [Progresses in detecting small hepatocellular carcinoma with digital subtraction angiography, CT and MRI]. PMID- 14552735 TI - [Roles of hepatitis B virus and hepatitis C virus in hepato-carcinogenesis]. PMID- 14552736 TI - Ability of Latin America laboratories to detect antimicrobial resistance patterns: experience of the SENTRY Antimicrobial Surveillance Program (1997 2000). AB - The accuracy of antimicrobial susceptibility tests is a crucial step for the clinical management of patients with serious infections. They must be reliable and precise because they will guide antimicrobial therapy. Our main objective was to compare the results of susceptibility testing performed by the SENTRY coordinator laboratory with those reported by the participating Latin American medical centers. A total of 10,277 bacterial isolates were tested by the reference broth microdilution method at the coordinator laboratory in the United States. The tests were performed and interpreted following the National Committee for Clinical Laboratory Standards (NCCLS) recommendations. Ten antimicrobial agent-organism combinations were analyzed. The susceptibility methods utilized in each of the medical centers were also evaluated. Total agreement of the results was obtained in nearly 88% of the antimicrobial agent-organism combinations. "Very major" (false-susceptible results) and "major errors" (false-resistant results) were observed in 12% and 6% of the cases, respectively. The highest disagreements were observed for coagulase-negative Staphylococcus_oxacillin (20% very major error) and Burkholderia cepacia_imipenem (21% - very major error). The susceptibility method with the highest agreement rate was Etest (92%) > PASCO (91%) > agar dilution (91%) > MicroScan (90%) > Vitek (87%). External quality assurance data obtained by surveillance programs such as the SENTRY Antimicrobial Surveillance Program are not only helpful for detecting the emergence of patterns of antimicrobial resistance, but also to monitor the performance of the participating microbiology laboratories. PMID- 14552737 TI - Effectiveness of the actions of antimicrobial control in the intensive care unit. AB - There are various strategies to improve the effectiveness of antibiotics in hospitals. In general, for the implementation of guidelines for appropriate antibiotic therapy, the participation of infectious disease (ID) physicians deserves considerable attention. This study was a prospective ecological time series study that evaluates the effectiveness of the ID physician's opinion to rationalize and control the use of antibiotics in medical-surgical intensive care units (ICU), and the impact of their intervention on treatment expenditures. There was significant change in the pattern of use of antimicrobials, this pattern approximating that of a medical-surgical ICU that participates in the ICARE (Intensive Care Antimicrobial Resistance Epidemiology) Project. For example, there was a significant increase in the consumption of antimicrobials of the ampicillin group (Relative Risk [RR]=3.39; 95% CI: 2.34-4.91) and antipseudomonal penicillins (RR=2.89; 95% CI: 1.70-4.92). On the other hand, there was a significant reduction in the consumption of 3(rd)/4(th )generation cephalosporins (RR=0.66; 95% CI: 0.57-0.77) and carbapenems (RR=0.43; 95% CI: 0.33-0.56). On average, for every patient-day antibiotic expense was reduced 37.1% during calendar year 2001, when compared with 2000. The ID specialists' opinion and the adoption of guidelines for empirical antibiotic therapy of hospital-acquired pneumonia contributed to a reduction in the use of antimicrobials in medical-surgical ICU. However, further studies that have more control over confounding variables are needed to help determine the relevance of these discoveries. PMID- 14552738 TI - Detection of Group A beta-hemolytic Streptococcus employing three different detection methods: culture, rapid antigen detecting test, and molecular assay. AB - In order to study the prevalence of Group A beta-hemolytic Streptococcus (GABHS) pharyngotonsillitis in our pediatric population and to compare different sampling methods of GABHS detection, oropharyngeal swabs from 50 children with acute pharyngotonsillitis, between 1 and 12 years old, were used simultaneously for culture, molecular assay and rapid GABHS antigen detection tests. All children were clinically examined at the Division of Pediatric Otorhinolaryngology of the Federal University of Sao Paulo. Diagnostic criteria were based on signs and symptoms, including sore throat, fever and oropharyngeal purulent secretion. Children that had been treated with antibiotics were excluded. Overall, combining the three methods, the prevalence of GABHS was 34%. GABHS was diagnosed in 30% of the bacterial cultures, in 25% of the samples tested with the molecular nucleic acid hybridization method and in 26% of the cases tested with the rapid antigen detection test. There was no significant difference between these three methods. PMID- 14552739 TI - Functional assesment of patients with AIDS disease. AB - We documented the types and degree of functional disability in 74 patients with AIDS at the Hospital de Clinicas of the Federal University of Parana, Brazil. Few of these patients are referred for rehabilitation services and there is only a limited team approach in their care. We found that 91% of the patients had some degree of functional impairment; in 81% the complaint was weakness and in 47% it was neurological involvement. According to the Barthel Index, 79% were considered functionally independent, 14% partially dependent and 7% dependent. For most of them, independence requires effort and their quality of life is reduced. Severe disability was rather unusual, while mild or moderate disability levels were not. The functional physical disability found in 21% of the patients required management by a rehabilitation team. We conclude that it is important to focus attention on the quality of life of patients with AIDS. PMID- 14552740 TI - An adherence trilogy is essential for long-term HAART success. AB - Adherence is the milestone of a successful therapy. Over the last decade several authors have addressed the importance of adherence for optimal results of antiretroviral (ARV) therapy. Many health care systems are investing substantial resources to make available contemporary antiretroviral therapy. Despite the large investment in medications, insufficient investments have been made into an integrated adherence component to maximize the impact of these medications. Adherence, unlike drug therapy, cannot be defined as a single method with a defined prescription or formula. Instead, it is the result of a complex interaction between the patient, a prescribed medication and the health system. Many reports are available analyzing each of these components. We have found that critical elements of adherence include the patient's knowledge about the disease and how medications will help achieve a longer and healthier life, together with the motivation to adapt to a new style of life. A trilogy composed of information, motivation and behavioral skills is essential to achieve the maximum desired level of adherence. We have computerized this trilogy in a software program for self-administration in which each of the three components is provided to the patient as many times as necessary to transmit an understanding of the problem and to help make a rational decision to adhere to the ARV treatment program. In this review we analyze several efforts and techniques to improve adherence to any recommended medication that may interfere with the patient's lifestyle and outline how the adherence trilogy can be best used to optimize the ability of ARV therapy to durably suppress plasma HIV RNA to undetectable levels. PMID- 14552741 TI - Progress towards meningitis prevention in the conjugate vaccines era. AB - Acute bacterial meningitis is an important cause of morbidity and mortality among children less than five years old, Haemophilus influenzae, Streptococcus pneumoniae and Neisseria meningitidis being the most important agents of bacterial meningitis in developing countries. The development of the conjugate vaccines in the beginning of the 90's, especially type b H. influenzae (Hib), and more recently the heptavalent pneumococcal and the serogroup C meningococcal vaccines, have contributed directly to changes in the epidemiological profile of these invasive diseases (direct effect) and of their carriage status (indirect effect). We review the impact of the Hib conjugate vaccine in Latin American countries, where this vaccine has been implemented, and the potential of pneumococcal and meningococcal conjugate vaccines for the reduction of meningitis worldwide. We also address constraints for the development and delivery of these vaccines and review new candidate state-of-the-art vaccines. The greatest challenge, undoubtedly, is to implement these vaccines worldwide, especially in the developing regions. PMID- 14552742 TI - Clinical presentation and follow up of children with congenital toxoplasmosis in Brazil. AB - We evaluated the clinical presentation and determined the ocular and neurologic sequelae in children with congenital toxoplasmosis in Brazil, taking into consideration the shortage of national publications on this disease. Follow-up evaluations were made of 43 children with congenital toxoplasmosis referred to Santa Casa de Sao Paulo, during a period of at least five years. Selection of the cases was based in clinical and laboratory criteria. A clear predominance of children with subclinical presentation of the disease at birth (88%) was found. Of the 43 children, 22 (51%) developed neurological manifestations. Using skull radiography, we detected neuroradiologic alterations in seven children (16%) and with tomography in 33 children (77%). Neurological sequelae were identified in 15 children (54%) in the group with cerebral calcifications and in 7 (47%) in the group without cerebral calcifications. We observed chorioretinitis in 95% of the cases. Reactivation of cicatricial lesions and the emergence of new ocular lesions were observed in five cases. The most frequent neurological manifestation was a delay in neuropsychomotor development. Most remarkable was the finding that cerebral calcifications were not associated with a higher incidence of neurological sequelae among the children. Chorioretinitis was the main ocular sequel of the infection, found in nearly all children; it can manifest years from birth, even in children submitted to specific therapy druing the first year of life, highlighting the importance of a follow-up of these children. PMID- 14552743 TI - Role of the Parasight-F test in the diagnosis of complicated Plasmodium falciparum malarial infection. AB - An evaluation was made of the diagnostic efficacy and utility of the Parasight-F test in diagnosing Plasmodium falciparum malaria, compared with conventional microscopy, particularly in severe and complicated cases. This study was designed as a prospective, case control hospital-based study. Febrile patients suspected to be suffering from malaria were selected randomly and were subjected to peripheral smear examinations (thick and thin) and Parasight-F tests till the required number of at least 30 cases of P. falciparum infection were identified, including at least 15 complicated cases. In addition 20 cases of P. vivax malarial infection as well as 20 healthy age and sex-matched individuals were taken as two control groups. The outcome measure was the number of cases with positive Parasight-F test results compared with conventional microscopy. Thirty two patients with P. falciparum malaria were identified, with 15 severe and complicated cases. Peripheral smears were positive in 29 (91%) of these, while parasight-F test was positive in 31 out of 32 (97%) cases. Parasites were detected only by bone marrow examination in one case. Diagnostic sensitivity and specificity of peripheral smears for detecting falciparum infection were 90.6% and 100% respectively while that of the Parasight-F test were 96.8% and 100%, respectively (P>.05). The Parasight-F test has high sensitivity and specificity in diagnosing P. falciparum malarial infection, comparable to or even higher than microscopy exams, particularly in severe and complicated cases, with additional advantages of speed, simplicity and objectivity. PMID- 14552744 TI - Seroprevalence and risk factors associated with Helicobacter pylori infection in blood donors in Salvador, Northeast-Brazil. AB - Helicobacter pylori plays an important role in the etiology of peptic ulcer disease. Its prevalence appears to be higher in developing countries. We evaluated the seroprevalence of H. pylori and risk factors associated with infection in voluntary blood donors who attended the main blood center of the city of Salvador, Brazil. The subjects responded to an epidemiological questionnaire, with information about sex, age, race, lifestyle, social-economic level indicators, and residence and hygiene conditions. Anti-H. pylori antibody was determined by ELISA (Cobas Core, Roche). Three hundred and seven subjects were included in the study. Anti-H. pylori antibody results were indeterminate in 33 individuals (10.8%), who were excluded from analysis. Among the remaining 274 subjects, 187 (68.2%) were anti-H. pylori positive. Based on multivariate logistic regression analysis three variables were found to be significantly associated with a higher prevalence of H. pylori infection: absence of plumbing in the residence during childhood, a history of rainwater invading the dwelling during childhood, and low ingestion of milk. PMID- 14552745 TI - Measles in children with HIV infection: report of five cases. AB - Patients with AIDS have a high incidence of skin problems due to the immunosuppression and malnourishment that are inherent to the progression of this disease. Clinical manifestation of these skin lesions and their severity are different in AIDS patients. We made a prospective study of five cases of measles in children with HIV infection during a community outbreak, and there were typical as well as atypical forms of the disease, including one case with negative serology. There were pulmonary complications, but none of the patients died. The anti-retroviral treatment may have contributed to the decrease in measles morbidity in these pediatric AIDS patients. PMID- 14552746 TI - Therapy with interferon-alpha plus ribavirin for membranoproliferative glomerulonephritis induced by hepatitis C virus. AB - We describe a treatment made with interferon-alpha (IFN-alpha) plus ribavirin of two patients with membranoproliferative glomerulonephritis (MPGN) induced by hepatitis C virus (HCV): case # 1 was a 22-yr-old woman with leg and facial edema, hypertension and proteinuria, whose liver biopsy revealed chronic active hepatitis; and case # 2 was a 42-yr-old man with anasarca, hypertension and proteinuria, whose liver biopsy indicated cirrhosis. Both had anti-HCV, HCV-RNA and cryoglobulins. IFN-alpha (3 million units (MU), 3 times/week) and ribavirin (1 g/day) were administered for 12 months. The drugs were well tolerated by both patients. Serum alanine aminotransferase (ALT) levels normalized and HCV-RNA became negative. Cryoglobulins disappeared and an improvement in renal disease was seen after 6 months of therapy. However, after 9 months, case # 2 presented ALT elevation, and proteinuria was detected. Two years after the end of therapy, both patients were negative in repeated HCV-RNA and cryoglobulin tests. Case # 1 was asymptomatic, with normal liver and renal tests, and case # 2 had normal blood pressure, with mild edema of the ankles. Based on the evolution of these two cases, the association of IFN-alpha and ribavirin may be a therapeutic option for patients with MPGN related to HCV. PMID- 14552748 TI - Design and synthesis of a new fluorescent probe for cytochrome P450 3A4 (CYP 3A4). AB - Inhibition of CYP 3A4 catalytic activity is a principal mechanism for in vivo drug-drug interactions, sometimes leading to severe toxic effects. Rapid in vitro testing for CYP 3A4 high affinity/high inhibition potential has become part of the standard investigations for new drug candidates. Unfortunately, the complexity of the kinetics associated with CYP 3A4 catalyzed reactions (multiple substrates binding, non Michaelis-Menten kinetics) make these tests either inaccurate or tedious. We have designed and synthesized a new fluorescent probe, a testosterone substituted at the 6beta- position with a fluorescent deazaflavine moiety which is able to inhibit to the same extent the hydroxylation of compounds known to bind to different sites in the CYP 3A4 active site. Furthermore, the binding of this compound and its displacement from the active site can be followed by fluorescence measurements, which allows a rapid evaluation of the CYP 3A4 affinity of any new drug candidate. PMID- 14552749 TI - Novel chromene derivatives as TNF-alpha inhibitors. AB - A novel series of chromene-based TNF-alpha inhibitors is described. These chromene derivatives inhibit bacterial lipopolysaccharide (LPS) stimulated production of TNF-alpha from human peripheral blood mononuclear cells (PBMC). Additionally, these compounds inhibit NF-kB mediated transcription activation. PMID- 14552750 TI - Identification of potent and novel small-molecule inhibitors of caspase-3. AB - The design and synthesis of a series of novel, reversible, small molecule inhibitors of caspase-3 are described. PMID- 14552751 TI - Synthesis and SAR studies of potent HIV protease inhibitors containing novel dimethylphenoxyl acetates as P2 ligands. AB - Isopropyl substituted 4-thioazolyl valine side chains are highly optimized P(2) P(3) ligands for C2 symmetry-based HIV protease inhibitors, as exemplified by the drug ritonavir. Replacement of the side chain with the conformationally constrained hexahydrofurofuranyloxy P(2) ligand in combination with a dimethylphenoxyacetate on the other end of the ritonavir core diamine yielded highly potent HIV protease inhibitors. The in vitro antiviral activity in MT4 cells increased by 10- and 20-fold, respectively, in the absence and presence of 50% human serum compared to ritonavir. The structure-activity relationships of inhibitor series with this combination of ligands were investigated. Preliminary pharmacokinetic studies in rats indicated rapid elimination of the inhibitors from the blood, and the plasma levels were not significantly enhanced by coadministration with ritonavir. However, the novel structural features and the high intrinsic antiviral potency of this series provides potential for the future exploration of prodrug strategies. PMID- 14552752 TI - Pyrazino[1,2-a]indole-1,4-diones, simple analogues of gliotoxin, as selective inhibitors of geranylgeranyltransferase I. AB - Some pyrazino[1,2-a]indole-1,4-diones, structurally simplified analogues of the natural mycotoxin gliotoxin, have been synthesised and investigated as inhibitors of prenyltransferases; one compound, 3-acetylthio-9-methoxy-2-methyl-2,3 dihydropyrazino[1,2-a]indole-1,4-dione 10 shows slightly greater selectivity (8 fold) for geranylgeranyltransferase type I (GGTase I) than gliotoxin itself. PMID- 14552753 TI - Growth inhibition activity of thioacetal artemisinin derivatives against human umbilical vein endothelial cells. AB - Thioacetal artemisinin derivatives, in particular, 10alpha phenylthiodihydroartemisinins (5), 10beta-benzenesulfonyl-9-epi dihydroartemisinin (9) and 10alpha-mercaptodihydroartemisinin (11), exhibit good growth inhibition activity against HUVEC proliferation at the concentration level of 1 microM. PMID- 14552754 TI - Phosphonooxymethyl prodrugs of the broad spectrum antifungal azole, ravuconazole: synthesis and biological properties. AB - Synthesis of phosphonooxymethyl derivatives of ravuconazole, 2 (BMS-379224) and 3 (BMS-315801) and their biological evaluation as potential water-soluble prodrugs of ravuconazole are described. The phosphonooxymethyl ether analogue 2 (BMS 379224) and N-phosphonooxymethyl triazolium salt 3 (BMS-315801) were both prepared from ravuconazole (1) and bis-tert-butyl chloromethylphosphate, but under two different conditions. Both derivatives were highly soluble in water and converted to the parent in alkaline phosphatase, and also in vivo (rat). However, BMS-315801 was found to be less stable than BMS-379224 in water at neutral pH. BMS-379224 (2) has proved to be one of the most promising prodrugs of ravuconazole that we tested, and it is currently in clinical evaluation as an intravenous formulation of the broad spectrum antifungal azole, ravuconazole. PMID- 14552755 TI - Nortropinyl-arylsulfonylureas as novel, reversible inhibitors of human steroid sulfatase. AB - Steroid sulfatase (STS) has emerged as an attractive target for a range of estrogen- and androgen-dependent diseases. Searching for novel chemotypes as STS inhibitors, we identified nortropinyl-arylsulfonylurea 3 as a hit from high throughput screening. A series of analogues was prepared in order to explore the essential structural elements for STS inhibition, and first structure-activity relationships were established. Mechanistic investigations revealed that the compounds are reversible, competitive inhibitors of STS. PMID- 14552756 TI - Time-dependence and preliminary SAR studies in inhibition of nitric oxide synthase isoforms by homologues of thiocitrulline. AB - Treatment of N(alpha)-Cbz-N(epsilon)-(2-hydroxyethylaminothiocarbonyl)-L-lysine N (2-hydroxyethyl)amide with boiling hydrochloric acid gave N(epsilon)-(4,5 dihydrothiazol-2-yl)-L-lysine. This was a weak and non-isoform selective inhibitor of NOS, whereas N(epsilon)-aminothiocarbonyl-L-lysine and its methyl ester were potent, with IC(50)=13 and 18 microM, respectively, against human iNOS and IC(50)=3 and 8 microM, respectively, against rat nNOS. Time dependence was observed for inhibition of nNOS by the ester. PMID- 14552758 TI - Biphenyl-based analogues of thiolactomycin, active against Mycobacterium tuberculosis mtFabH fatty acid condensing enzyme. AB - Analogues of the antibiotic thiolactomycin, with biphenyl-based 5-substituents, were found to have excellent in vitro inhibitory activity against the recombinant Mycobacterium tuberculosis beta-ketoacyl-ACP synthase mtFabH condensing enzyme. In particular, 5-(4'-benzyloxy-biphen-4-ylmethyl)-4-hydroxy-3,5-dimethyl-5H thiophen-2-one exhibited approximately a 4-fold increased potency against this key condensing enzyme involved in M. tuberculosis mycolic acid biosynthesis, compared to thiolactomycin. PMID- 14552757 TI - Biological evaluation of sphingomyelin analogues as inhibitors of sphingomyelinase. AB - Seeking neutral sphingomyelinase inhibitors, we designed and synthesized hydrolytically stable analogues of sphingomyelin. These novel analogues replace the phosphodiester moiety of sphingomyelin with carbamate and urea moiety, resulting in inhibition of neutral sphingomyelinase. Compound 1 prevented ceramide generation and apoptotic neuronal cell death in a model of ischemia based on organotypic hippocampal slice cultures. PMID- 14552759 TI - Styrylheterocycles: a novel class of inhibitors on lipopolysaccharide-Induced nitric oxide production. AB - A series of styrylheterocycles was prepared and their inhibitory activities against nitric oxide (NO) production were evaluated in a cell culture system using lipopolysaccharide-stimulated RAW264.7 macrophage cells. Our studies have identified a new series of inhibitors on NO production, providing the basis for further development of potent inhibitors. The preliminary structure-activity relationship, to elucidate the essential structural requirements, has been described. Mechanistic studies suggest that the suppression of iNOS mRNA transcription is, at least in part, related to the inhibitory activity of styrylheterocycles. PMID- 14552760 TI - Identification of novel inhibitors of BCR-ABL tyrosine kinase via virtual screening. AB - Inhibition of BCR-ABL tyrosine kinase activity has shown to be essential for the treatment of chronic myelogenous leukemia (CML). However, drug resistance has quickly arisen in recent clinical trials for STI571 (Gleevec), which is the first approved drug of CML by inhibiting ABL tyrosine kinase. It is desirable to develop new types of ABL tyrosine kinase inhibitors that may overcome this drug resistance problem. Here we present the discovery of novel inhibitors targeted at the catalytic domain of ABL tyrosine kinase by using three-dimensional database searching techniques. From a database containing 200,000 commercially available compounds, the top 1000 compounds with the best DOCK energy score were selected and subjected to structural diversity and drug likeness analysis, 15 compounds were submitted for biological assay. Eight out of the 15 showed inhibitory activity against K562 cells with IC(50) value ranging from 10 to 200 microM. Two promising compounds showed inhibition in further ABL tyrosine phosphorylation assay. It is anticipated that those two compounds can serve as lead compounds for further drug design and optimization. PMID- 14552761 TI - Precursor-Directed polyketide biosynthesis in Escherichia coli. AB - Precursor-directed polyketide biosynthesis was demonstrated in the heterologous host Escherichia coli. Several diketide and triketide substrates were fed to a recombinant E. coli strain containing a variant form of deoxyerythronolide B synthase (DEBS) from which the first elongation module was deleted resulting in successful macrolactone formation from the diketide, but not the triketide, substrates. PMID- 14552763 TI - Low molecular mass peptide dendrimers that express antimicrobial properties. AB - A series of low-generation dendrimeric peptides was synthesized in an attempt to evaluate their antimicrobial potency. All tested dendrimeric peptides in which lysine was a starting and branching element expressed moderate activity against Staphylococcus aureus NCTC 4163, and Escherichia coli NCTC 8196. PMID- 14552762 TI - Identification of a novel class of inhibitor of human and Escherichia coli thymidine phosphorylase by in silico screening. AB - Structure-based computational screening of the National Cancer Institute database of anticancer compounds identified novel non-nucleobase-derived inhibitors of human thymidine phosphorylase as candidates for lead optimization. The hierarchical in silico screening strategy predicted potentially strong low molecular weight ligands exhibiting a range of molecular scaffolds. Of the thirteen ligands assayed for activity, all displayed inhibitory activity against Escherichia coli thymidine phosphorylase. One compound, hydrazine carboxamide 2 [(1-methyl-2,5-dioxo-4-pentyl-4-imidazolidinyl)methylene], was found to inhibit E. coli thymidine phosphorylase with an IC(50) value of 20 microM and an IC(50) value of 77 microM against human thymidine phosphorylase. As this hydantoin derivative lacks the undesirable ionic sites of existing tight-binding nucleobase derived inhibitors, such as 5-chloro-6-[(2-iminopyrrolidin-1-yl)methyl]uracil hydrochloride, it provides an opportunity for the design of potent thymidine phosphorylase inhibitors with improved pharmacokinetic properties. PMID- 14552764 TI - A general synthesis of 1-aryl carbamoyl-2-alkyl-4-aryl substituted semicarbazides as nonbasic factor Xa inhibitors. AB - An efficient four-step synthesis of 1-aryl-carbamoyl-2-alkyl-4-aryl semicarbazides starting from benzophenone hydrazone is described leading to moderately active neutral factor Xa inhibitors. PMID- 14552765 TI - Practical, asymmetric synthesis of 16-hydroxyeicosa-5(Z),8(Z), 11(Z),14(Z) tetraenoic acid (16-HETE), an endogenous inhibitor of neutrophil activity. AB - An asymmetric synthesis of 16-HETE, an endogenous inhibitor of neutrophil activity, was achieved in six steps from R-(-)-glycidyl benzyl ether in 28% overall yield. PMID- 14552766 TI - Synthesis and X-ray crystal structures of substituted fluorobenzene and benzoquinone inhibitors of the tissue factor VIIa complex. AB - Multistep syntheses of substituted benzenes and benzoquinone inhibitors of tissue Factor VIIa are reported. The benzene analogues were designed such that their substitution pattern would occupy and interact with the S(1), S(2), and S(3) pockets of the tissue Factor VIIa (TF/VIIa) enzyme. The compounds exhibited modest potency on TF/VIIa with selectivity over Factor Xa and thrombin. The X-ray crystal structures of the targeted fluorobenzene 12a and benzoquinone 14 inhibitors bound to TF/VIIa were obtained and will be described. PMID- 14552767 TI - A novel class of apical sodium co-dependent bile acid transporter inhibitors: the 1,2-benzothiazepines. AB - A series of 5-aryl-3,3-dibutyl-7-(dimethylamino)-1,2-benzothiazepin-4-ol 1,1 dioxides were prepared and were found to inhibit the apical sodium co-dependent bile acid transporter (ASBT) for the potential treatment for hyperlipidemia. Several 1,2-benzothiazepines exhibited low nanomolar in vitro activity. The synthesis and initial in vitro potency data is presented for this novel class of compounds. PMID- 14552768 TI - Synthesis of beta-substituted cationic porphyrins and their interactions with DNA. AB - The beta-substituted cationic porphyrins (7, 8 and 10) have been synthesized and their interactions with plasmid DNA investigated. We found that substituents at the beta-position of porphyrins (7 and 8) have apparently affected their interactions with DNA compared with non-beta-substituted porphyrins (10). PMID- 14552769 TI - Substrate properties of C5-substituted pyrimidine 2'-deoxynucleoside 5' triphosphates for thermostable DNA polymerases during PCR. AB - In order to enhance a collection of modified deoxynucleoside triphosphates useful for in vitro selection or SELEX (systematic evolution of ligands by exponential enrichment) techniques, we designed and synthesized modified analogues of 2' deoxyuridine triphosphate and 2'-deoxycytidine triphosphate bearing a flexible and hydrophilic 7-amino-2,5-dioxaheptyl linker at a C5 position. Both analogues were found to be substrates for thermostable DNA polymerases which belong to an evolutional family B during PCR. PMID- 14552770 TI - Regioselective synthesis and cytotoxicities of camptothecin derivatives modified at the 7-, 10- and 20-positions. AB - A series of 7-acyloxymethylcamptothecin and 20-O-acyl-7-acyloxymethylcamptothecin derivatives were regioselectively prepared on different solvents. 7 Acyloxymethylcamptothecins possess more efficacy than 20-O-acyl-7 acyloxymethylcamptothecins against six human cancer cell lines in vitro. PMID- 14552771 TI - Novel non-peptide inhibitors targeting death receptor-mediated apoptosis. AB - We have previously reported that ECH, (2R, 3R, 4S)-2,3-epoxy-4-hydroxy-5 hydroxymethyl-6-(1E)-propenyl-cyclohex-5-en-1-one inhibits Fas-mediated apoptosis by blocking self-activation of pro-caspase-8 in the death-inducing signaling complex (DISC). A series of ECH derivatives were asymmetrically synthesized via key synthetic intermediates obtained from lipase-catalyzed kinetic resolution. Inhibitory activities of the derivatives towards death receptor-mediated apoptosis both in type I and type II cells were investigated, revealing that novel non-peptide inhibitors, RKTS-33 and RKTS-34, are effective as ECH. PMID- 14552772 TI - QSAR study on antibacterial activity of sulphonamides and derived Mannich bases. AB - The paper describes synthesis and comparative study on antibacterial activities of sulphonamides and Mannich bases derived from them. The compounds were screened for their antibacterial activity against various gram-positive and gram-negative bacteria and were analyzed statistically. The results have shown that the compounds are quiet active against pathogens under study and were nontoxic. PMID- 14552773 TI - Exploring selectivity requirements for COX-2 versus COX-1 binding of 3,4 diaryloxazolones using E-state index. AB - Considering the importance of developing selective COX-2 inhibitors, the present paper explores selectivity requirements for COX-2 versus COX-1 binding of 3,4 diaryloxazolones using electrotopological state (E-state) index. The study also shows the utility of E-state index in developing statistically acceptable model having direct physicochemical significance: electron density distribution of different atoms of the oxazolone ring and attached two phenyl rings are important for the selective binding with COX-2 over COX-1. Moreover, the use of indicator variable shows that presence of ortho R(1) substituent (except fluoro) on the N(3)-phenyl ring decreases COX-2 selectivity. Further, an amino substituent at R(2) position (i.e., sulfonamide compound) is favorable for increasing COX-2 selectivity when the R(3) position is unsubstituted. PMID- 14552774 TI - The total synthesis of an aurone isolated from Uvaria hamiltonii: aurones and flavones as anticancer agents. AB - The naturally occurring aurone 1, isolated from Uvaria hamiltonii, and a series of aurones analogues based structurally on known tubulin binding agents were prepared and evaluated for anticancer activity. Aurone 20 was the most active (IC(50) K562 50 nM) and caused significant G2/M cell-cycle arrest. PMID- 14552775 TI - Synthesis and biological evaluation of GABA derivatives able to cross the blood brain barrier in rats. AB - Two new GABA derivatives, 1 and 2, were synthesized and tested for their capacity to display CNS activity, which was assessed by determining the effects on the duration of pentobarbital-induced hypnosis in rats. Compound 1, peripherally injected, significantly prolonged the hypnosis time, a typical GABA-mimetic effect, while both intracerebroventricular and intravenous administration of compound 2 surprisingly shortened the hypnotic effect in an atropine-sensitive way. The study was extended also to compounds 1a, 1b and 2a, putative oxidative/hydrolytic metabolites of 1 and 2. PMID- 14552776 TI - New benzylidenethiazolidinediones as antibacterial agents. AB - A novel benzylidenethiazolidinedione has been discovered with antimicrobial activity. Here, we present the results of a structure-activity study on this compound with respect to its antimicrobial activity. PMID- 14552777 TI - Synthesis of a highly active new anti-HIV agent 2',3'-didehydro-3'-deoxy-4' ethynylthymidine. AB - Compounds having methyl, vinyl, and ethynyl groups at the 4'-position of stavudine (d4T: 2',3'-didehydro-3'-deoxythymidine) were synthesized. The compounds were assayed for their ability to inhibit the replication of HIV in cell culture. The 4'-ethynyl analogue (15) was found to be more potent and less toxic than the parent compound stavudine. PMID- 14552778 TI - The acute EPS of haloperidol may be unrelated to its metabolic transformation to BCPP+. AB - We have previously proposed that haloperidol's debilitating extrapyramidal symptoms (EPS) may be associated with its quaternary BCPP+ (an MPP+ like species) metabolite formed in vivo. However, recent work on D2 knock out mice suggests that haloperidol's EPS may be related to its potent D2 binding (K(i)=0.9 nM). In this study, we explore this question by synthesizing and testing an analogue (DS 27) that binds to D2 receptors with higher affinity than haloperidol, but cannot form quaternary metabolites. This study suggests that D2 affinity may be the primary underlying mechanism for acute catalepsy induction by haloperidol. PMID- 14552779 TI - Synthesis and antimalarial evaluation of new 1,4-bis(3-aminopropyl)piperazine derivatives. AB - Synthesis and evaluation of the activity of a new family of 1,4-bis(3 aminopropyl)piperazine derivatives against a chloroquine-resistant strain of Plasmodium falciparum, and as inhibitors of beta-hematin formation, are described. The highest antimalarial activities were obtained for compounds displaying the highest predicted vacuolar accumulation ratios and the best potencies as inhibitors of beta-hematin formation. The most potent compound displayed an activity 3-fold better than chloroquine for a comparable selectivity index upon MRC-5 cells. Therefore, in this series, the replacement of the 7 chloroquinoline group can constitute a strong rationale for further investigation. PMID- 14552780 TI - A theoretical investigation on DPPH radical-scavenging mechanism of edaravone. AB - The mechanism of edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one) to scavenge DPPH radical is clarified by density functional theory (DFT) calculations. It is revealed that H-atom-abstraction rather than electron-transfer reaction is involved in the radical-scavenging process of edaravone, and H-atom at position 4 is readily to be abstracted. The C-H bond dissociation enthalpy (BDE) of edaravone is higher than the O-H BDE of alpha-tocopherol, accounting for the activity difference between the two antioxidants. As substituents have little influence on the C-H BDE, 2-pyrazolin-5-one is recognized as the active center for edaravone. PMID- 14552781 TI - Aryl piperazine melanocortin MC4 receptor agonists. AB - Incorporation of substituted phenyl piperazine privileged structures into a known MC4 specific dipeptoid consensus sequence resulted in a series of potent (EC(50)=24 nM) and selective MC4-R agonists. We report the SAR of this series of compounds using in vitro cAMP functional assays in cells transfected with the MC4 or other melancortin receptors. PMID- 14552782 TI - Investigation of the effect of varying the 4-anilino and 7-alkoxy groups of 3 quinolinecarbonitriles on the inhibition of Src kinase activity. AB - Several 7-alkoxy-4-anilino-3-quinolinecarbonitriles were synthesized and evaluated for Src kinase inhibitory activity. Optimal inhibition of both Src enzymatic and cellular activity was seen with analogues having a 2,4-dichloro-5 methoxyaniline group at C-4. Compound 18, which has a 1-methylpiperidinemethoxy group at C-7, showed in vivo activity in a xenograft model. PMID- 14552783 TI - Determination of binding affinities of triplex forming oligonucleotides using a fluorescent intercalator displacement (FID) assay. AB - The binding affinities of several triplex forming oligonucleotides were determined using a fluorescent intercalator displacement (FID) assay. PMID- 14552784 TI - High-resolution assessment of protein DNA binding affinity and selectivity utilizing a fluorescent intercalator displacement (FID) assay. AB - Protein titration displacement of ethidium bromide bound to hairpin deoxyoligonucleotides containing any sequence of interest provides a well-defined titration curve (measuring the loss of fluorescence derived from the DNA bound ethidium bromide) that provides both absolute binding constants (K(a)) and stoichiometry of binding. This use of a fluorescent intercalator displacement (FID) assay for establishing protein DNA binding affinity and selectivity is demonstrated with the examination of the LEF-1 HMG domain binding to hairpin deoxyoligonucleotides containing its commonly accepted consensus sequence 5' CTTTGWW (W=A or T) and those modified (5'-CTNTGWW) to examine sequences implicated in early studies (5'-CTNTG). The effectiveness of the FID assay coupled with its technically non-demanding experimental use makes it an attractive alternative or complement to selection screening, footprinting or affinity cleavage, and electrophoretic mobility shift assays for detecting, characterizing, and quantitating protein DNA binding affinity and selectivity. PMID- 14552785 TI - Photoregulation of deacylation rate of acyl trypsin derived from photoresponsive inverse substrate. AB - The acyl trypsin was prepared by use of an inverse substrate, which is comprise of a photoresponsive 4-phenylazobenzoyl moiety. The acyl group in acyl trypsin has been shown to isomerize from trans-form (4t-trypsin) to cis-form (4c trypsin)/from cis-form to trans-form by irradiation of UV-vis light. The deacylation rate of the cis-form (4c-trypsin) has been shown to be 18.6 times faster than that of the trans-form (4t-trypsin). PMID- 14552786 TI - Structure-activity relationship of triaryl propionic acid analogues on the human EP3 prostanoid receptor. AB - Potent and selective ligands for the human EP3 prostanoid receptor are described. Triaryl compounds bearing an ortho-substituted propionic acid moiety were identified as potent EP3 antagonists based on the SAR described herein. The binding affinities of key compound on all eight human prostanoid receptors is reported. PMID- 14552787 TI - A novel series of histone deacetylase inhibitors incorporating hetero aromatic ring systems as connection units. AB - A series of structurally novel HDAC inhibitors, in which a hetero aromatic ring connects the spacer with the hydrophobic group, has been designed and synthesized. These new inhibitors are very potent in in vitro enzymatic assays and display antiproliferation activity against two human cancer cell lines. PMID- 14552789 TI - Structure elucidation of Sch 538415, a novel acyl carrier protein synthase inhibitor from a microorganism. AB - A novel acyl carrier protein synthase inhibitor, Sch 538415 (1), was isolated from an unidentified bacterial microbe. Structure elucidation of 1 was accomplished based on analysis of spectroscopic data including UV, MS and 2D-NMR spectra. Compound 1 exhibited inhibitory activity in the acyl carrier protein synthase (AcpS) assay with an IC(50) value of 4.19 microM and showed antibacterial activity against Staphylococcus aureus in the agar diffusion assay. PMID- 14552788 TI - Synthesis and biological evaluation of 4-[(3-methyl-3H-imidazol-4-yl)-(2 phenylethynyl-benzyloxy)-methyl]-benzonitrile as novel farnesyltransferase inhibitor. AB - Farnesyltransferase inhibitors (FTIs) have emerged as a novel class of anticancer agents. Analogues of the potent FTI, 4-[3-biphenyl-1-hydroxy-1-(3-methyl-3H imidazol-4-yl)-prop-2-ynyl]-1-yl-benzonitrile, were synthesized and tested in vitro for their inhibitory activities. The most promising compound identified from this series is analogue 11 that possesses potent enzymatic and cellular activities. PMID- 14552790 TI - Ganglioside GM1 mimics: lipophilic substituents improve affinity for cholera toxin. AB - Ganglioside GM1 mimics including (R)-2-hydroxy-3-cyclohexylpropionic acid or (R) 2-hydroxy-3-phenylpropionic acid as replacements for NeuAc are stronger cholera toxin binders than the parent ligand 2, which includes (R)-2-hydroxy-propionic acid. PMID- 14552791 TI - Aryl[a]pyrrolo[3,4-c]carbazoles as selective cyclin D1-CDK4 inhibitors. AB - The synthesis of new analogues of Arcyriaflavin A in which one indole ring is replaced by an aryl or heteroaryl ring is described. These new series of aryl[a]pyrrolo[3,4-c]carbazoles were evaluated as inhibitors of Cyclin D1-CDK4. A potent and selective D1-CDK4 inhibitor, 7a (D1-CDK4 IC(50)=45 nM), has been identified. The potency, selectivity profile against other kinases, and structure activity relationship (SAR) trends of this class of compounds are discussed. PMID- 14552792 TI - Studies on cyclin-dependent kinase inhibitors: indolo-[2,3-a]pyrrolo[3,4 c]carbazoles versus bis-indolylmaleimides. AB - A series of indolo[2,3-a]pyrrolo[3,4-c]carbazoles and their bis-indolylmaleimides precursors have been prepared in order to compare their activity as D1-CDK4 inhibitors. Both enzymatic and antiproliferative assays have shown that the structurally more constrained indolo[2,3-a]pyrrolo[3,4-c]carbazoles are consistently more active (8-42-fold) in head-to-head comparison with their bis indolylmaleimides counterparts. Cell-cycle analysis using flow cytometry have also shown that the indolocarbazoles are selective G1 blockers while the bis indolylmaleimides arrest cells in the G2/M phase. PMID- 14552793 TI - Rigidified acetylcholine mimics: conformational requirements for binding to neuronal nicotinic receptors. AB - Rigidified derivatives have been designed and synthesized assuming the g+t conformer of acetylcholine (N-C-C-O=+60 degrees, C-C-O-C=180 degrees ) as active conformation for binding to cytisine sensitive neuronal nicotinic receptors. The SAR of the compounds evaluated, along with those of more flexible analogues, support the g+t conformer hypothesis and highlight the stringent steric limitation of this nicotinic receptor sub-type. Compound 3e has low microM affinity for cytisine sensitive nicotinic receptor binding sites while being selective with regard to the alpha-bungarotoxin sensitive subclass. We also report few compounds with microM affinity for the alpha-bungarotoxin sensitive subclass. PMID- 14552794 TI - The C-4 stereochemistry of leucocyanidin substrates for anthocyanidin synthase affects product selectivity. AB - Anthocyanidin synthase (ANS), an iron(II) and 2-oxoglutarate (2OG) dependent oxygenase, catalyses the penultimate step in anthocyanin biosynthesis by oxidation of the 2R,3S,4S-cis-leucoanthocyanidins. It has been believed that in vivo the products of ANS are the anthocyanidins. However, in vitro studies on ANS using optically active cis- and trans-leucocyanidin substrates identified cyanidin as only a minor product; instead both quercetin and dihydroquercetin are products with the distribution being dependent on the C-4 stereochemistry of the leucocyanidin substrates. PMID- 14552795 TI - Indole-2-carboxamides as novel NR2B selective NMDA receptor antagonists. AB - A novel series of indole-2-carboxamide derivatives was prepared and identified as NR2B selective NMDA receptor antagonists. The influence of the number and position of OH groups on the indole skeleton as well as the substitution of the piperidine ring on the biological activity of the compounds was studied. PMID- 14552796 TI - Enantiomerically pure tetrahydroquinoline derivatives as in vivo potent antagonists of the glycine binding site associated to the NMDA receptor. AB - To identify neuroprotective agents after stroke, new substituted tetrahydroquinoline derivatives were designed as antagonists of the glycine binding site associated to the NMDA receptor, satisfying the key pharmacophoric requirements. In particular, the racemate 3c exhibited outstanding in vivo activity in the MCAo model in rats, when given iv both pre- and post-ischemia. Pure enantiomers 3c-(+) and 3c-(-) have been prepared following an original synthetic route. Despite the significant difference of activity observed in vitro, they shown similar neuroprotective profile in the MCAo model in rats. PMID- 14552797 TI - Synthesis and evaluation of chiral bicyclic proline FKBP12 ligands. AB - As part of our ongoing program to explore novel structural classes of FKBP12 ligands, we herein wish to report a new class of FKBP12 ligands containing chiral bicyclic proline analogues. Details of the synthetic routes, together with preliminary biological activity, will be presented. PMID- 14552798 TI - X-ray structures of two xanthine inhibitors bound to PEPCK and N-3 modifications of substituted 1,8-dibenzylxanthines. AB - The analysis of the X-ray structures of two xanthine inhibitors bound to PEPCK and a comparison to the X-ray structure of GTP bound to PEPCK are reported. The SAR at N-1, N-7 and developing SAR at C-8 are consistent with information gained from the X-ray structures of compounds 1 and 2 bound to PEPCK. Representative N-3 modifications of compound 2 that led to the discovery of 3-cyclopropylmethyl and its carboxy analogue as optimal N-3 groups are presented. PMID- 14552799 TI - Sulphonamide-based small molecule VLA-4 antagonists. AB - The discovery of a sulphonamide by-product with VLA-4 antagonistic activity led to a series of potent, small molecule VLA-4 antagonists. Synthesis, SAR and in vivo evaluation of the selected compound will be presented. PMID- 14552801 TI - How is complex second-order motion processed? AB - Converging psychophysical and electrophysiological evidence suggests that first order (luminance-defined) complex motion types i.e., radial and rotational motion, are processed by specialized extrastriate motion mechanisms. We ask whether radial and rotational second-order (texture-defined) motion patterns are processed in a similar manner. The motion sensitivity to translating, radiating and rotating motion patterns of both first-order (luminance-modulated noise) and second-order (contrast-modulated noise) were measured for patterns presented at four different exposure durations (106, 240, 500 and 750 ms). No significant difference in motion sensitivity was found across motion type for the first-order motion class across exposure duration (i.e., from 240 to 750 ms) whereas direction-identification thresholds for radiating and rotating second-order motion were significantly greater than that of the second-order translational stimuli. Furthermore, thresholds to all second-order motion stimuli increased at a significantly faster rate with decreasing exposure duration compared to those of first-order motion. Interestingly, simple and complex second-order thresholds increased at similar rates. Taken together, the results suggest that complex second-order motion is not analyzed in a sequential manner. Rather, it seems that the same 'hard-wired' mechanisms responsible for complex first-order motion processing also mediate complex second-order motion, but not before the pre processing (i.e., rectification) of local second-order motion signals. PMID- 14552802 TI - Experience-dependent visual cue recalibration based on discrepancies between visual and haptic percepts. AB - We studied the hypothesis that observers can recalibrate their visual percepts when visual and haptic (touch) cues are discordant and the haptic information is judged to be reliable. Using a novel visuo-haptic virtual reality environment, we conducted a set of experiments in which subjects interacted with scenes consisting of two fronto-parallel surfaces. Subjects judged the distance between the two surfaces based on two perceptual cues: a visual stereo cue obtained when viewing the scene binocularly and a haptic cue obtained when subjects grasped the two surfaces between their thumb and index fingers. Visual and haptic cues regarding the scene were manipulated independently so that they could either be consistent or inconsistent. Experiment 1 explored the effect of visuo-haptic inconsistencies on depth-from-stereo estimates. Our findings suggest that when stereo and haptic cues are inconsistent, subjects recalibrate their interpretations of the visual stereo cue so that depth-from-stereo percepts are in greater agreement with depth-from-haptic percepts. In Experiment 2 the visuo haptic discrepancy took a different form when the two surfaces were near the subject than when they were far from the subject. The results indicate that subjects recalibrated their interpretations of the stereo cue in a context sensitive manner that depended on viewing distance, thereby making them more consistent with depth-from-haptic estimates at all viewing distances. Together these findings suggest that observers' visual and haptic percepts are tightly coupled in the sense that haptic percepts provide a standard to which visual percepts can be recalibrated when the visual percepts are deemed to be erroneous. PMID- 14552803 TI - Monocular transparency generates quantitative depth. AB - Monocular zones adjacent to depth steps can create an impression of depth in the absence of binocular disparity. However, the magnitude of depth is not specified. We designed a stereogram that provides information about depth magnitude but which has no disparity. The effect depends on transparency rather than occlusion. For most subjects, depth magnitude produced by monocular transparency was similar to that created by a disparity-defined depth probe. Addition of disparity to monocular transparency did not improve the accuracy of depth settings. The magnitude of depth created by monocular occlusion fell short of that created by monocular transparency. PMID- 14552804 TI - Mental extrapolation of target position is strongest with weak motion signals and motor responses. AB - Some accounts hold that the position of moving objects is extrapolated either in visual perception or visual short-term memory ("representational momentum"). However, some studies did not find forward displacement of the final position when smooth motion was used, whereas reliable displacement was observed with implied motion. To resolve this conflict, the frequency of position changes was varied to sample motion types between the extreme cases of implied and smooth motion. A continuous function relating frequency of target change and displacement was found: Displacement increased when the frequency of position changes was reduced. Further, the response mode was varied. Probe judgments produced less forward displacement than motor judgments such as mouse or natural pointing movements. Also, localization judgments were susceptible to motion context, but not to variations of probe shape or expectancy about trajectory length. It is suggested that forward displacement results from the extrapolation of the next step in the observed motion sequence. PMID- 14552805 TI - Contrast conservation in human vision. AB - Visual experience, which is defined by brief saccadic sampling of complex scenes at high contrast, has typically been studied with static gratings at threshold contrast. To investigate how suprathreshold visual processing is related to threshold vision, we tested the temporal integration of contrast in the presence of large, sudden changes in the stimuli such occur during saccades under natural conditions. We observed completely different effects under threshold and suprathreshold viewing conditions. The threshold contrast of successively presented gratings that were either perpendicularly oriented or of inverted phase showed probability summation, implying no detectable interaction between independent visual detectors. However, at suprathreshold levels we found complete algebraic summation of contrast for stimuli longer than 53 ms. The same results were obtained during sudden changes between random noise patterns and between natural scenes. These results cannot be explained by traditional contrast gain control mechanisms or the effect of contrast constancy. Rather, at suprathreshold levels, the visual system seems to conserve the contrast information from recently viewed images, perhaps for the efficient assessment of the contrast of the visual scene while the eye saccades from place to place. PMID- 14552806 TI - Peak localization of sparsely sampled luminance patterns is based on interpolated 3D surface representation. AB - Objects in the world are typically defined by contours and local features separated by extended featureless regions. Sparsely sampled profiles were therefore used to evaluate the cues involved in localizing objects defined by such separated features (as opposed to typical Vernier acuity or other line-based localization tasks). Objects, in the form of Gaussian blobs, were defined at the sample positions by luminance cues, binocular disparity cues or both together. Remarkably, the luminance information in the sampled profiles was unable to support localization for objects requiring interpolation when the perceived depth from the luminance cue was cancelled by a disparity cue. Disparity cues, on the other hand, improved localization substantially over that for luminance cues alone. These data indicate that it is only through the interpolated depth representation that the position of the sampled object can be recognized. The dominance of a depth representation in the performance of such tasks shows that the depth information is not just an overlay to the 2D sketch of the positional information, but a core process that must be completed before the position of the object can be recognized. PMID- 14552807 TI - Extending the shine-through effect to classical masking paradigms. AB - A vernier, presented for a short time, shines through a following grating if the grating contains nine and more elements but remains largely invisible for smaller gratings. Therefore, extended grating masks yield, surprisingly, less masking than smaller ones. Here, we show that this mask size effect is not unique to grating masks. Masking diminishes if the size of classical pattern-, noise-, light-, and metacontrast masks increases and if these masks are regular, i.e. highly ordered. PMID- 14552808 TI - Sequential memory-guided saccades and target selection: a neural model of the frontal eye fields. AB - We present a neural model of the frontal eye fields. It consists of several retinotopic arrays of neuron-like units that are recurrently connected. The network is trained to make memory-guided saccades to sequentially flashed targets that appear at arbitrary locations. This task is interesting because the large number of possible sequences does not permit a pre-learned response. Instead locations and their priority must be maintained in active working memory. The network learns to perform the task. Surprisingly, after training it can also select targets in visual search tasks. When targets are shown in parallel it chooses them according to their salience. Its search behavior is comparable to that of humans. It exhibits saccadic averaging, increased reaction times with more distractors, latency vs accuracy trade-offs, and inhibition of return. Analysis of the network shows that it operates like a queue, storing the potential targets in sequence for later execution. A small number of unit types are sufficient to encode this information, but the manner of coding is non obvious. Units respond to multiple targets similar to quasi-visual cells recently studied [Exp. Brain Res. 130 (2000) 433]. Predictions are made that can be experimentally tested. PMID- 14552809 TI - Effects of stimulus size and luminance on oscillopsia in congenital nystagmus. AB - Although the absence of oscillopsia is a common feature of congenital nystagmus (CN), it is occasionally noted by patients under poor viewing conditions and has been provoked in laboratory settings with stabilised images. In the present study, the effects of reductions in background stimulus size and luminance on perceptual stability in CN were examined. Sixteen CN subjects were first interviewed using a structured questionnaire about whether they ever experienced oscillopsia and, if so, under what circumstances and with what perceptions. They next fixated an LED centred in projected images of three sizes (21x14 degrees, 10x6 degrees and 7x4 degrees) and four luminance levels (115.5, 24.5, 2.7 and 0.1 cd/m2, with contrasts from 96 down to 20%). Eye movements were recorded with a limbal tracker. They were asked after viewing each image "whether anything happened to the image while they watched it." Occasional oscillopsia was reported by 12/16 of the CN subjects on the questionnaire. In the laboratory, 13/16 subjects experienced oscillopsia in some manner for at least one of the stimuli. 8/13 CN subjects experienced it for the dimmest and smallest slides. 11/13 perceived certain parts (either the LED or background) of the visual stimuli as moving, with the perception of LED movement most pronounced at low background luminance. Foveation did not differ when trials with and without reported oscillopsia were compared (independent samples t-test, p>0.05). Oscillopsia may occur in CN with normal viewing of bright fixation targets against dim backgrounds. Under these conditions, the oscillopsia may be spatially inhomogeneous. Luminance differences between the fixation point and surround may have caused transmission time differences as the image moved across the retina, therefore leading to the perception of motion in one portion of the scene and not the other. PMID- 14552810 TI - Contrast coding and magno/parvo segregation revealed in reaction time studies. AB - Reaction times (RTs) are obtained for a wide range of contrasts of vertical sinusoidal gratings. The data are plotted as a function of the reciprocal of contrast. In some conditions, a single linear function accounts for the data. In others a clear bi-linear function is obtained. The low and high contrast regions of the function are interpreted as representing magno and parvo activity, respectively. RT-based supra-threshold sensitivity functions are obtained for different luminances, stimulus durations and eccentricities and these are compared with conventional threshold-based sensitivities to establish the extent to which RTs and contrast sensitivity are constrained by the same sensory processes. PMID- 14552811 TI - Procedure for separation of GM2 ganglioside species with different ceramide structures by a flash reversed-phase silica gel liquid chromatography. AB - GM2 ganglioside, beta-GalNAc-(1-4)-[alpha-Neu5Ac-(2-3)-]beta-Gal-(1-4)-beta-Glc (1-1)-Cer, is the main ganglioside in the brain of Tay-Sachs patients. In this work, GM2 ganglioside was extracted from a Variant B Tay-Sachs human brain, purified to homogeneity of the oligosaccharide moiety by silica gel chromatography. It was further fractionated for the first time into the molecular species differing in the ceramide structures by reverse-phase flash chromatography. The GM2 ganglioside species were characterized by gas chromatography, nuclear magnetic resonance spectroscopy, and mass spectrometry. The major GM2 species contained the ceramides with d18:1-18:0 (40.5% of the total GM2 species), d20:1-18:0 (31%) and d18:1-20:0 (12%). We also found minor GM2 species with the ceramides with d18:1-24:1 (4%), d18:1-22:0 (2%) and d18:2-24:1 (1%), which have not been reported previously. PMID- 14552812 TI - Determination of isoprostanes in urine samples from Alzheimer patients using porous graphitic carbon liquid chromatography-tandem mass spectrometry. AB - F2-isoprostanes (F2-iPs) comprise four classes of isomers produced non enzymatically by free radical attack on arachidonic acid, a component of the cell membrane. This paper describes a new method for the quantification of F2 isoprostanes in urine samples from thoroughly diagnosed Alzheimer's disease (AD) patients. The sample pretreatment consisted of liquid extraction of 900 microl urine with diethyl ether, its subsequent evaporation, and finally, reconstitution in 50 microl water. Of this, 20 microl was injected into a HPLC system with a 15 mm x 1 mm porous graphitic carbon column coupled to a triple quadrupole mass spectrometer running in negative electrospray ionization mode. The F2 isoprostanes were separated in 15 min using a linear solvent gradient comprising water, methanol, acetonitrile and ammonium hydroxide at a pH of 9.5. The average recovery obtained was approximately 75%. The limit of detection (3S/N) was calculated for iPF2alpha-III to be 0.7 pg injected on column, corresponding to 0.1 nM. The average level of iPF2alpha was 241 +/- 163 pg/mg creatinine in the urine samples from AD patients (average +/- standard deviation). The corresponding control values were 216 +/- 101 pg/mg creatinine, i.e. no statistically significant difference was noticed. No correlation pattern specific to Alzheimer's disease was revealed by principal component analysis of the isoprostane peaks obtained either. The results from this study support earlier findings that levels of peripheral isoprostanes are not increased in patients with Alzheimer's disease. PMID- 14552813 TI - Simultaneous determination of major B-trichothecenes and the de-epoxy-metabolite of deoxynivalenol in pig urine and maize using high-performance liquid chromatography-mass spectrometry. AB - A selective analytical method based on high-performance liquid chromatography (HPLC), combined with atmospheric pressure chemical ionisation (APCI-) mass spectrometry (MS), has been developed for simultaneous determination of B trichothecenes and the major metabolites of deoxynivalenol. The method allows simultaneous analysis of nivalenol (NIV), deoxynivalenol (DON), 15 acetyldeoxynivalenol (15-AcDON), 3-acetyldeoxynivalenol (3-AcDON), fusarenon X (Fus-X) and de-epoxydeoxynivalenol (DOM-1). The method is based on one-step sample clean-up using a multifunctional MycoSep column. A linear gradient mobile phase system, consisting of water:acetonitrile:methanol (H2O:ACN:MeOH) at a flow rate of 1 ml/min, and a Polar-RP C18 column, were utilised to obtain the best resolution of all tested compounds along with column and equilibrating within 30 min. Dexamethasone (Dex) was used as internal standard. The developed method shows good repeatability for inter- and intra-day precisions as well as good linearity of calibration curves (r2 ranged from 0.9936 to 0.9998). Average recoveries for tested compounds in both matrices have been determined ranging from 63.7 to 102.3% and limit of quantification (LOQ) ranged from 25 to 150 ng/g. The utility and practical impact of the method is demonstrated using contaminated pig urine and maize samples. PMID- 14552814 TI - Chiral liquid chromatography resolution and stereoselective pharmacokinetic study of the enantiomers of a novel anticonvulsant, N-(4-chlorophenyl)-1-(4 pyridyl)ethylamine, in rats. AB - A selective chiral high performance liquid chromatographic (HPLC) method was developed and validated to separate and quantify the enantiomers of a novel anticonvulsant agent, N-(4-chlorophenyl)-1-(4-pyridyl)ethylamine (AAP-Cl), in rat plasma. After extraction of the plasma samples with ethyl acetate, the separation was accomplished by an HPLC system consisting of a Chirex chiral column (250 mm x 4.6 mm i.d.) and a mobile phase of hexane:ethanol:tetrahydrofuran (280:20:40 (v/v)) containing trifluroacetic acid (0.3% (v/v)) and triethylamine (0.018% (v/v)) at a flow rate of 0.8 ml/min with UV detection. Male Sprague-Dawley rats were given (+)-AAP-Cl (10 and 20 mg/kg), (-)-AAP-Cl (10 mg/kg) or the racemic mixture (20 mg/kg) by i.v. bolus injection and serial blood samples were collected at different times after drug administration. (+)-AAP-Cl and (-)-AAP-Cl were separated with a resolution factor, Rs, of at least 1.4, and a separation factor, alpha, greater than 1.09. Linear calibration curves were obtained over the concentration range of 0.5-30 microg/ml in plasma for both (+)-AAP-Cl and (-) AAP-Cl (R2 > or = 0.996) with a limit of quantitation of 100 ng/ml and the recovery was greater than 80% for both enantiomers. The accuracy and precision for both enantiomers ranged from 96 to 102% (+/-0.2-7%) at upper and lower concentrations. The plasma concentration-time profiles of the enantiomers of AAP Cl were best described by a two-compartment open model with a mean terminal half life of about 5h, volume of distribution at steady state of 3 l/kg and clearance of about 0.6l/(hkg) in rats. There was no significant difference between the pharmacokinetic parameters of (+)-AAP-Cl and (-)-AAP-Cl, suggesting that the disposition of AAP-Cl in rats is not enantioselective. In addition, no chiral inversion of (+)-AAP-Cl to (-)-AAP-Cl or vice versa was observed. The results of this investigation have shed some light on the mechanism of action and disposition of AAP-Cl in rats. PMID- 14552815 TI - Enantioselective liquid chromatographic-electrospray mass spectrometric assay of beta-adrenergic blockers: application to a pharmacokinetic study of sotalol in human plasma. AB - An enantioselective high performance liquid chromatographic-electrospray ionization mass spectrometric (HPLC-ESI-MS) method for the direct determination of several beta-adrenergic blockers was developed and validated. The method is based on the direct separation of the enantiomers of drugs on a laboratory-made chiral stationary phase (CSP) containing covalently bonded teicoplanin (TE) as chiral selector. Detection of the effluent was performed by electrospray ionization mass spectrometry, run in the selected-ion recording (SIR) mode. The method was applied to the pharmacokinetic monitoring of sotalol (STL) in the plasma of five young healthy volunteers, dosed with racemic drug. The limits of quantitation (LOQ) reached 4 ng/ml for both sotalol enantiomers. Such a method, fully validated, offers a novel, fast and very efficient tool for the direct determination of sotalol enantiomers in human plasma, and can be generally applied to the beta-adrenergic blockers stereoselective pharmacokinetics. PMID- 14552816 TI - Identification of pheromones in mouse urine by head-space solid phase microextraction followed by gas chromatography-mass spectrometry. AB - Given the key role of pheromones in animal communication and behaviour, there is need to identify the different classes of these molecules under varying physiological conditions. However, the highly volatile nature of pheromones and the fact that they occur at very low concentrations in urine makes this task all the more difficult. Herein, we present a method of detecting and identifying the five main pheromones known: 2-sec-butyl-4,5-dihydrothiazole, geraniol, indole, trans-beta farnesene and trans-alpha farnesene in individual urine microsamples taken from male mice. Urine volumes as small as 20 microl were subjected to solid phase microextraction (SPME) followed by gas chromatography-mass spectrometry (GC MS). This selective analytical method permits the rapid detection of these pheromones free from cross-contaminants as a clearly distinguishable spectral signals. Highest recovery rates of natural pheromones were achieved by extraction on a carboxen/polydimethylsiloxane (CAR/PDMS) fibre of 85 microm film thickness. This selective, sensitive and accurate method will help address the question of possible links between certain pheromone classes, and social and reproductive behaviour in mice. PMID- 14552817 TI - Assay of leucine aminopeptidase activity in vitro using large-pore reversed-phase chromatography with fluorescence detection. AB - A chromatographic method for determination of leucine aminopeptidase (LAP) activity in complex matrices is described. L-Leucine-beta-naphthylamide was used as the substrate and its hydrolytic product, beta-naphthylamine, was monitored by fluorescence at 280 nm excitation and 400 nm emission wavelengths. Under optimized conditions, the components in the incubation mixture were baseline separated and eluted out of a large-pore (300 angstroms) reversed-phase C4 column (RPC4) within 15 min with a non-linear gradient elution of methanol (0.05% (v/v) trifluoroacetic acid additive). The detection limit of the hydrolytic product reached 0.35 pmol at three time signal-to-noise (S/N) ratio with 5 microl sample injection. The method showed a wide dynamic range for quantitation of both the hydrolytic product (10 ng/ml to 80 microg/ml) and LAP (0.1-46.0 microg/ml) with correlation coefficient larger than 0.998 and reproducibility <3 and 10% R.S.D. (n=3), respectively. A fairly broad range of incubation time could be selected within 1 h. The LAP activities and concentrations in rabbit serum, tears, and mouse lens homogenates were determined to be 41.8 (0.3 mg/ml), 2.8 (40.0 microg/ml), and 1.6 pmol/(microl min) (17.5 microg/ml), respectively, with reproducibility of 2-9% R.S.D. (n=3) and intra- and inter-day variation for the retention time of the hydrolytic product being <1% R.S.D. (n=3). The results indicate that the present method is rapid and sensitive as compared to the conventional one. PMID- 14552818 TI - Determination of rat oral bioavailability of soy-derived phytoestrogens using an automated on-column extraction procedure and electrospray tandem mass spectrometry. AB - In recent years, consumption of herbal supplements as an alternative to pharmaceutical drug therapy has increased. For example, with the health claims labeling which describes the link between soy-protein and a reduced risk of coronary heart disease (CHD), the consumption of soy and soy-derived phytoestrogens has increased dramatically. That being said, the oral bioavailability of only a few soy phytoestrogens such as Daidzein and Genestein have been previously estimated. In this paper, we present the calculated percent of rat oral bioavailability of five soy-derived phytoestrogens (Genistein, Daidzein, Biochanin A, Coumestrol, and Zearalenone) in male Sprague-Dawley rats. The plasma quantitation required for the bioavailability calculation is performed by using a rapid on-line plasma extraction procedure for the quantitative analysis. To further speed up the analysis the rats were dosed using the 'n-in one' (cassette) protocol. The rapid on-line extraction/quantitation methodology coupled to the cassette dosing analysis of phytoestrogens is the key point of this paper. The limit of quantitation (LOQ) for each compound was 1-1000 ng/ml with each plasma sample analysis taking less than 2 min. In general the percent oral bioavailability was determined to be between 11 and 28%. PMID- 14552819 TI - High-performance liquid chromatographic assay of metabolites of thioguanine and mercaptopurine in capillary blood. AB - The main metabolites of the cytotoxic drugs thioguanine (6TG) and mercaptopurine (6MP) can be measured conveniently in red blood cells (RBC). Isolation of RBC, however, is laborious and requires some milliliters of blood. This HPLC assay allows measurements of thiopurine metabolites in very small blood samples obtained from the finger-tip. The metabolites, derivatives of 6TG and methylmercaptopurine (6MeMP), were extracted and hydrolized with perchloric acid to liberate the corresponding base. 6MeMP is completely transformed under these conditions to 4-amino-5-(methylthio)carbonyl imidazole. The chromatographic separation of 6TG and this imidazole was performed in a single run under isocratic conditions within 10 min using a 70 mm column. The quantification limit was 0.5 nmol/ml for 6TG and 3 nmol/ml blood for 6MeMP. The accuracy was 83% for 6TG (CV=3%) over the concentration range of 0.5-20 nmol/ml blood and 102% (CV=4%) for 6MeMP over the range of 3-150 nmol/ml blood. The intra-assay CV ranged from 5.4 to 7.4% for 6TG and from 6.2 to 10.6% for 6MeMP. The inter-assay CV was 7.5 and 9.5% in a pooled blood sample. The levels in RBC in whole blood were nearly coincident with those obtained in separated RBC, isolation of RBC therefore is not necessary for these measurements, if the drugs are given per os in the day before blood sampling. The concentration of 6MeMP nucleotides is more dependent on the given 6MP dose than the concentration of 6TG nucleotides. Intraindividual variations were small at unchanged drug doses, interindividual metabolite concentrations were highly variable. PMID- 14552820 TI - Determination of morphine and morphine glucuronides in human plasma by 96-well plate solid-phase extraction and liquid chromatography-electrospray ionization mass spectrometry. AB - There is considerable interest in quantifying morphine and its major metabolites, morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G). Available assays use gas chromatography-mass spectrometry or high-performance liquid chromatography (HPLC) with single or tandem mass spectrometry, ultraviolet, electrochemical, or fluorimetric detection. Nevertheless, few methods provide adequate sensitivity for all analytes, in a single injection, with the desired rate of sample throughput. A rapid and sensitive method for quantification of morphine, M3G and M6G from human plasma using HPLC with electrospray ionization mass spectrometry was developed using a Waters Oasis MCX 96-well plate for extracting both lipophilic morphine and its hydrophilic glucuronides, C18 separation using an isocratic mobile phase (methanol, acetonitrile and formic acid), and selected ion monitoring. Recoveries of morphine, M3G and M6G, respectively, were 81, 90 and 82% at the low (2, 25 and 2 ng/ml), 80, 77 and 75% at the medium (10, 250 and 10 ng/ml), and 74, 62 and 72% at the high (100, 1000 and 100 ng/ml) quality control samples. The limit of quantitation was 0.5 ng/ml morphine and M6G, and 5 ng/ml M3G. Analytes were validated over a linear range of 0.5-200 ng/ml morphine and M6G, and 5-2000 ng/ml M3G. This assay represents an improvement over existing methods through solid phase extraction with increased sample throughput (96-well plates), use of small samples (0.5 ml), and sub nanogram detection. PMID- 14552821 TI - Improved high-performance liquid chromatographic method for the pharmacokinetic studies of a novel iron chelator, CP502, in rats. AB - An improved reverse-phase high-performance liquid chromatographic method (RP HPLC) for the determination of a novel iron chelator CP502 (1,6-dimethyl-3 hydroxy-4-(1H)-pyridinone-2-carboxy-(N-methyl)-amide hydrochloride) in rat plasma, urine and feces was developed and validated. The separation was performed on a polymeric column using a mobile phase composed of 1mM ethylenediaminetetra acetic acid disodium salt (EDTA), acetonitrile, methanol and methylene chloride. Separation of CP502 from plasma, urine or feces endogenous compounds was achieved by gradient elution. Retention times of CP502 and its major metabolite (glucuronide) were about 13 and 4 min, respectively. The method was validated in terms of limit of detection (LOD), limit of quantification (LOQ), selectivity (endogenous from plasma, urine or feces), linearity, extraction recovery, robustness (column selection, mobile phase composition, detection mode, internal standard (IS) selection, analyte stability), day-to-day reproducibility and system suitability (repeatability, peak symmetry and resolution). The method is applicable to bioavailability and pharmacokinetic studies of CP502 in rats. PMID- 14552822 TI - High-performance liquid chromatographic assay for the determination of Aloe Emodin in mouse plasma. AB - An isocratic high-performance liquid chromatography (HPLC) method was developed and validated to determine Aloe Emodin (AE) in mouse plasma. The analysis required 0.3 ml of plasma and involves extraction with dichloromethane. The HPLC separation was carried out on Symmetry Shield RP18, a mobile phase of methanol water-acetic acid (65:35:0.2) and fluorescence detection at lambda(ex)=410 nm and lambda(em)=510 nm. The retention time of AE was 11.7 min. The assay was linear from 10 to 1,000 ng/ml (r2 > or = 0.999), showed intra- and inter-day precision within 7.8 and 4.7%, and accuracy of 87.3-105.7%. Detection limit (LOD) and quantification limit (LOQ) were 4.5 and 5 ng/ml, respectively. The method was applied to determine for the first time the pharmacokinetic of AE in mice. PMID- 14552823 TI - Simultaneous determination of androstenediol 3-sulfate and dehydroepiandrosterone sulfate in human serum using isotope diluted liquid chromatography-electrospray ionization-mass spectrometry. AB - A simple method for simultaneous determination of androstenediol 3-sulfate (Adiol 3S) and dehydroepiandrosterone sulfate (DHEA-S) in human serum using isotope diluted liquid chromatography-electrospray ionization-ion trap-mass spectrometry (LC-ESI-ion trap-MS) was developed. After addition of deuterated internal standards ([2H5]Adiol-3S and [2H4]DHEA-S), human serum (100 microl) was deproteinized with acetonitrile and then applied to a solid-phase extraction cartridge, Oasis HLB. The obtained steroid sulfates fraction was washed with hexane and then analyzed by LC-ESI-MS operated in the negative ion mode. The quantification ranges of Adiol-3S and DHEA-S were 10-400 ng/ml and 0.05-8 microg/ml, respectively. The method does not require the chemical or enzymatic hydrolysis of the conjugates and purification with high-performance liquid chromatography, and shows satisfactory reproducibility and accuracy. The concentrations of these sulfates in the sera of healthy male volunteers (n=14) were 19.2-245.3 mg/ml (Adiol-3S) and 0.175-5.16 microg/ml (DHEA-S), and those of patients with prostate cancer (n=19) were 15.3-182.7 ng/ml (Adiol-3S; four samples, not detectable) and 0.110-2.421 microg/ml (DHEA-S). PMID- 14552824 TI - Determination of the major mercapturic acids of 1,3-butadiene in human and rat urine using liquid chromatography with tandem mass spectrometry. AB - The major urinary metabolites of 1,3-butadiene are monohydroxybutenyl-mercapturic acids (MHBMA) and dihydroxy-butyl-mercapturic acid (DHBMA). These metabolites can be used as biomarkers of exposure to this diene. In order to determine the smoking-related exposure to 1,3-butadiene, we have developed a rapid LC-MS/MS method for the determination of MHBMA and DHBMA in urine of humans and rats. The method requires 2-5 ml of urine which is solid phase extracted prior to LC-MS/MS analysis. Precision for MHBMA is < or =11.2% for human and < or = 17% for rat urine. Corresponding values for DHBMA are < or = 7.2 and < or = 19%, respectively. Recovery rates are approximately 100% for both analytes in human urine and about 115% in rat urine. Limits of detection (LOD) are for humans 0.9 and 23 ng/ml and for rats 1.5 and 33 ng/ml for MHBMA and DHBMA, respectively. Application of the method to urine of humans and rats showed a significant effect of tobacco smoke exposure on the urinary excretion of MHBMA and the metabolic ratio DHBMA/(DHBMA + MHBMA). PMID- 14552825 TI - Search for peptidic "middle molecules" in uremic sera: isolation and chemical identification of fibrinogen fragments. AB - According to the "middle molecule" (MM) hypothesis, the uremic solutes ranging from 500 to 5,000 Da are insufficiently eliminated by conventional hemodialysis and may act as uremic toxins. However, because of the methodological difficulties of MM purification, their chemical analysis is complicated and the precise structure of these molecules remains obscure. In the present study, a new micro preparative procedure including SDS electrophoresis and liquid chromatography was applied for isolation of MM peptides from uremic sera. Microsequencing and MS/MS analyses of these peptides showed that most of the identified MM (22 out of 23) represented the N- and C-terminal fragments of the alpha- and beta-chains of fibrinogen. The obtained data provide new information on the precise structure of fibrinogen fragments accumulating in uremic serum as MM. PMID- 14552826 TI - Development of a liquid chromatography method for the determination of linezolid and its application to in vitro and human microdialysis samples. AB - Linezolid is a new, promising antibacterial agent to treat severe infections. A rapid HPLC assay using UV detection for the determination in microdialysate and human plasma was developed. After sample preparation, using acetonitrile for plasma and water for microdialysate, 20 microl was injected and separated on a RP 18 column. Overall, the assay exhibited good precision and accuracy. The diffusion properties of linezolid investigated in in vitro microdialysis experiments revealed a mean relative recovery of 77.5% (CV: 5.4%; delivery and recovery experiments). Following characterization of linezolid in in vitro microdialysis, the setting is suitable for application in clinical studies. PMID- 14552827 TI - Reliable and specific high-performance liquid chromatographic method for simultaneous determination of loratadine and its metabolite in human plasma. AB - A high-performance liquid chromatographic (HPLC) method with fluorescence detection has been developed for the simultaneous determination of loratadine (L) and its metabolite, descarboethoxyloratadine (DCL), in human plasma. Following a two-step liquid-liquid extraction with toluene, the analytes were separated using a gradient mobile phase consisting of methanol-acetonitrile-phosphate buffer. The linearity for L and DCL was within the concentration range of 0.5-16 ng/ml. The coefficient of variation of intra- and inter-day assay was <8.3%, with accuracy ranging from 98.3 to 105.7%. The lower limit of quantification was 0.5 ng/ml for both L and DCL. This method has been demonstrated to be reliable, and is an improvement over existing methods due to its capability for determining L and DCL simultaneously in a single chromatographic run. PMID- 14552828 TI - Measurement of nicotine and cotinine in human milk by high-performance liquid chromatography with ultraviolet absorbance detection. AB - A high-performance liquid chromatographic (HPLC) assay for the determination of nicotine and cotinine in human milk was developed using an extraction by liquid liquid partition combined with back extraction into acid, and followed by reverse phase chromatography with UV detection of analytes. The assay was linear up to 500 microg/l for both nicotine and cotinine. Intra- and inter-day relative standard deviations (R.S.D.) were <10% (25-500 microg/l) for both nicotine and cotinine. Limits of quantitation (LOQ) were 10 and 12 microg/l for nicotine and cotinine, respectively, while the limits of detection (LOD) were 8 and 10 microg/l for nicotine and cotinine, respectively. The mean recoveries were 79-93% (range 25-500 microg/l) for nicotine and 78-89% (range 25-500 microg/l) for cotinine. The amount of fat in the milk did not affect the recovery. We found that this method was sensitive and reliable in measuring nicotine and cotinine concentrations in milk from a nursing mother who participated in a trial of the nicotine patch for smoking cessation. PMID- 14552829 TI - Liquid-chromatographic determination of erlotinib (OSI-774), an epidermal growth factor receptor tyrosine kinase inhibitor. AB - A high-performance liquid-chromatographic (HPLC) assay with UV detection has been developed for the quantitative determination of erlotinib (OSI-774) in human plasma. Quantitative extraction was achieved by a single-solvent extraction involving a mixture of acetonitrile and n-butyl chloride (1:4, v/v). Erlotinib and the internal standard hydrochloride salt (OSI-597) were separated on a column packed with Nova-Pak C18 material and a mobile phase composed of acetonitrile and water, pH 2.0 (60:40, v/v). The column effluent was monitored with dual UV detection at wavelengths of 348 nm (erlotinib) and 383 nm (OSI-597). The calibration graph was linear in the range of 100-4500 ng/ml, with values for accuracy and precision ranging from 87.9 to 96.2% and 2.13 to 5.10%, respectively, for three different sets of quality control samples. The developed method was successfully applied to study the pharmacokinetics of erlotinib in a cancer patient at the recommended daily dose of 150 mg. PMID- 14552830 TI - Pharmacokinetic study of ellagic acid in rat after oral administration of pomegranate leaf extract. AB - Quantification of ellagic acid, the principal bioactive component of pomegranate leaf extract, in rats plasma following oral administration of pomegranate leaf extract was achieved by using a high-performance liquid chromatographic method. The calibration curve for ellagic acid was linear (r2=0.9998) ver the concentration range 0.026-1.3 microg/ml. The intra- and inter-day assays of ellagic acid from rat plasma were less than 6.52% at concentration range from 26 to 1300 ng/ml and good overall recoveries (94.5-102.4%) were found on same concentrations. The concentration-time profile was fitted with an open two compartment system with lag time and its max concentration of ellagic acid in plasma was 213 ng/ml only 0.55 h after oral administration extract 0.8 g/kg. The pharmacokinetic profile indicates that ellagic acid has poor absorption and rapid elimination after oral administration pomegranate leaf extract, and part of it was absorbed from stomach. PMID- 14552831 TI - Liquid chromatography determination of 10-hydroxycamptothecin in human serum by a column-switching system containing a pre-column with restricted access media and its application to a clinical pharmacokinetic study. AB - A simple, rapid, sensitive column-switching HPLC method is described for the analysis of the 10-hydroxycamptothecin (HCPT) in human serum. A pre-column containing restricted access media (RAM) is used for the sample clean-up and trace enrichment and is combined with a C18 column for the final separation. The analytical time is 8 min. The HCPT is monitored with fluorescence detector, excitation and emission wavelengths being 385 and 539 nm, respectively. There is a linear response range of 1-1000 ng/ml with correlation coefficient of 0.998 while the limit of quantification is 0.1 ng/ml. The intra-day and inter-day variations are less than 5%. This analytic procedure has been applied to a pharmacokinetic study of HCPT in clinical patients and the pharmacokinetic parameters of one-compartment model are calculated. PMID- 14552832 TI - Improved procedure for the determination of malonaldehyde by gas-chromatography with electron-capture detection as 2,4,6-trichlorophenylhydrazine derivative. AB - A previously described derivatization method using trichlorophenylhydrazine was developed for the estimation of malonaldehyde measured by gas-chromatography (GC) with electron-capture detection. The precision and reliability of the procedure are improved here by the use of methylmalonaldehyde as internal standard and by the introduction of a diverter valve at the end of the capillary column to protect the electron-capture detector, respectively. The method was applied to determine malonaldehyde content in bovine plasma samples. PMID- 14552833 TI - Differential signaling by CpG DNA in DCs and B cells: not just TLR9. AB - CpG-containing oligodeoxynucleotides (CpG ODNs) act on Toll-like receptor 9 (TLR9) that is expressed on B cells and plasmacytoid dendritic cells (pDCs) to stimulate the innate immune system, however, different types of CpG ODNs induce distinct responses. Recent papers suggest some CpG ODNs could require a second receptor or cofactor to signal. The different signaling complexes assembled might impact on the affinity with which CpG ODNs signal to TLR9 or activate additional pathways that lead to distinct immune responses. PMID- 14552834 TI - Self-renewal of hematopoietic and leukemic stem cells: a central role for the Polycomb-group gene Bmi-1. AB - Self-renewal of hematopoietic stem cells is vital for the sustained daily production of blood cells. Two recent studies have shown that the Polycomb-group gene Bmi-1 is indispensable for regulation of self-renewal by normal and leukemic stem cells. This identifies Polycomb-group genes as potential targets for therapeutic intervention in leukemia, and possibly other forms of cancer. PMID- 14552835 TI - BTLA: a new inhibitory receptor with a B7-like ligand. AB - The B and T lymphocyte attenuator, BTLA, is a recently discovered Ig superfamily member. BTLA engagement results in downregulation of T-cell activation, and mice deficient in BTLA show increased incidence and severity of autoimmune disorders. A novel B7 family member, B7x (B7S1, B7-H4), has been proposed as the ligand for BTLA. A B7x-Ig fusion protein inhibits T-cell activation and interleukin-2 (IL-2) production. Thus, cytotoxic T lymphocyte antigen-4 (CTLA-4), programmed death-1 (PD-1) and BTLA comprise an expanding set of inhibitory receptors expressed on lymphocytes. PMID- 14552837 TI - The interferon in TLR signaling: more than just antiviral. AB - The Toll-like receptor (TLR) system is responsible for the recognition of infectious agents leading to initiation of the primary innate, and later adaptive, immune response. Genetic technologies have enabled the discovery of new factors involved in these systems, their genetic manipulation and the global analyses of their effects on gene expression. Furthermore, this increased understanding has resulted in the need to reassess our preconceptions about the functions of well-known molecules. For example, type I interferons (IFNs), which were discovered as antiviral proteins, are now known to be produced in response to TLR activation by many pathogens, including bacteria. Should we be surprised? Has the inflammatory response unexpectedly highjacked the body's antiviral system? Or are we too easily blinkered by preconceptions from how a compound was discovered? PMID- 14552836 TI - Leucine-rich repeats and pathogen recognition in Toll-like receptors. AB - Toll-like receptors (TLRs) are the major cell-surface initiators of inflammatory responses to pathogens. They bind a wide variety of pathogenic substances through their ectodomains (ECDs). Here, we ask: what is the structural basis for this interaction? Toll-like receptor ECDs comprise 19-25 tandem copies of a motif known as the leucine-rich repeat (LRR). No X-ray structure of a TLR-ECD is currently available but there are several high-resolution LRR-containing proteins that can be used to model TLRs. We suggest that the basic framework of TLRs is a horseshoe-shaped solenoid that contains an extensive beta-sheet on its concave surface, and numerous ligand-binding insertions. Together, these insertions and the beta-sheet could provide a binding surface that is 10-fold greater in area than binding surfaces in antibodies and T-cell receptors. PMID- 14552838 TI - Introducing baselines for therapeutic use of regulatory T cells and cytokines in autoimmunity. AB - The concept of therapeutic immune regulation aiming to treat autoimmune diseases has been validated in multiple animal models, yet, the development of strategies for treatment of human autoimmune diseases remains problematic. Main obstacles are the contradictory findings in different model systems, as well as the contrasting functions of regulatory lymphocytes and cytokines. By drawing examples primarily from experimental type 1 diabetes, we propose that regulatory cells and cytokines can be classified according to the baseline at which they operate in healthy individuals and disease states that are not accompanied by severe systemic immune deficiency or skewing. Consequently, deletion or neutralization of regulatory cells or cytokines operative at high levels to maintain systemic homeostasis should constitute a therapeutic strategy for immune enhancement (e.g. tumor- and pathogen-specific immunity), whereas boosting these factors will have limited effects if the therapeutic goal is a downmodulation of immune responses (e.g. autoimmunity). Conversely, regulatory cells and cytokines operative at low homeostatic levels should unfold therapeutic capacities by further embellishment but not additional reduction. PMID- 14552839 TI - OCA-B regulation of B-cell development and function. AB - The transcriptional co-activator OCA-B [for Oct co-activator from B cells, also known as OBF-1 (OCT-binding factor-1) and Bob1] is not required for B-cell genesis but does regulate subsequent B-cell development and function. OCA-B deficient mice show strain-specific, partial blocks at multiple stages of B-cell maturation and a complete disruption of germinal center formation in all strains, causing humoral immune deficiency and susceptibility to infection. OCA-B probably exerts its effects through the regulation of octamer-motif controlled gene expression. The OCA-B gene encodes two proteins of distinct molecular weight, designated p34 and p35. The p34 isoform localizes in the nucleus, whereas the p35 isoform is myristoylated and is bound to the cytoplasmic membrane. p35 can traffic to the nucleus and probably activates octamer-dependent transcription, although this OCA-B isoform might regulate B cells through membrane-related signal transduction. PMID- 14552840 TI - T-cell receptor signal transmission: who gives an ITAM? AB - T cells have an amazing ability to discern and differentially respond to MHC embedded peptides that can differ by only a single amino acid. This potential involves a combination of the precise ligand-binding specificities of the T-cell receptor (TCR) and the distinct intracellular signaling processes it transmits. Signaling processes are controlled by the ten immunoreceptor tyrosine-based activation motifs (ITAMs) present in the invariant chains of the TCR complex (TCR zeta and CD3-gamma, -delta and -epsilon ). Here, we discuss recent studies of the functions of TCR invariant chains and the contribution of the ten ITAMs to T-cell signal transmission. We incorporate these results into two non-exclusive models of TCR signal transduction: the ITAM multiplicity model, which describes a functional redundancy within the TCR zeta and CD3 ITAMs; and the differential signaling model, which proposes distinct functions for the CD3-gamma, -delta and epsilon and TCR zeta modules. PMID- 14552841 TI - LAD-III, a novel group of leukocyte integrin activation deficiencies. AB - To extravasate the bloodstream at specific targets, circulating immune cells must activate their integrins to undergo rapid in situ modulation of affinity or avidity for their endothelial ligands. This activation involves specialized sub second G-protein signal transduction triggered by endothelium-displayed chemoattractants--primarily chemokines--and their cognate leukocyte-expressed G protein-coupled receptors (GPCRs). Recently, we reported a rare autosomal recessive leukocyte adhesion deficiency (LAD) syndrome associated with a defective ability of integrins to undergo GPCR-mediated stimulation at endothelial contacts. This LAD shows significant similarities to a group of integrin-activation syndromes reported in leukocytes and platelets. Here, the mechanisms by which GPCRs might regulate leukocyte and platelet integrins are outlined with respect to this new family of LAD cases. We propose to term this the LAD-III family. PMID- 14552843 TI - A framework for analyzing the cognitive complexity of computer-assisted clinical ordering. AB - Computer-assisted provider order entry is a technology that is designed to expedite medical ordering and to reduce the frequency of preventable errors. This paper presents a multifaceted cognitive methodology for the characterization of cognitive demands of a medical information system. Our investigation was informed by the distributed resources (DR) model, a novel approach designed to describe the dimensions of user interfaces that introduce unnecessary cognitive complexity. This method evaluates the relative distribution of external (system) and internal (user) representations embodied in system interaction. We conducted an expert walkthrough evaluation of a commercial order entry system, followed by a simulated clinical ordering task performed by seven clinicians. The DR model was employed to explain variation in user performance and to characterize the relationship of resource distribution and ordering errors. The analysis revealed that the configuration of resources in this ordering application placed unnecessarily heavy cognitive demands on the user, especially on those who lacked a robust conceptual model of the system. The resources model also provided some insight into clinicians' interactive strategies and patterns of associated errors. Implications for user training and interface design based on the principles of human-computer interaction in the medical domain are discussed. PMID- 14552844 TI - Using usability heuristics to evaluate patient safety of medical devices. AB - OBJECTIVE: To modify the traditional heuristic evaluation method of assessing software usability so that it can be applied to medical devices and used to evaluate the patient safety of those devices through the identification and assessment of usability problems. DESIGN: Heuristic evaluation, a usability inspection method commonly used for software usability evaluation, was modified and extended for medical devices. The modified method was used to evaluate and compare the patient safety of two 1-channel volumetric infusion pumps. RESULTS: The modified heuristic evaluation method was successfully applied to medical devices. One hundred and ninety-two heuristic violations were categorized for 89 usability problems identified for Pump 1, and 121 heuristic violations were categorized for the 52 usability problems identified for Pump 2. Pump 1 had more usability problems with high severity ratings than Pump 2. In general, Pump 1 was found to have more usability issues that are likely to induce more medical errors. CONCLUSIONS: Heuristic evaluation, when modified for medical devices, is a useful, efficient, and low cost method for evaluating patient safety features of medical devices through the identification of usability problems and their severities. PMID- 14552845 TI - Institutional decision-making to select patient care devices: identifying venues to promote patient safety. AB - Many medical errors that involve drug infusion devices are related to classic interface problems. Although manufacturers are becoming increasingly aware of human factors design considerations, many devices that are currently on the market are still sub-optimal for human use. This places significant responsibility for device selection on institutional purchasing groups. Theories of naturalistic decision-making point to many potential strengths and pitfalls of group decision-making processes that may affect the final outcome. This paper describes a retrospective analysis of decision-making process for infusion pump selection in a large hospital and focuses on factors related to patient safety. Through a series of detailed interviews and a study of relevant documentation we characterized the nature of the decision-making, patterns of communication, and the roles of different participants. Findings show that although the process involves a number of different professional groups and committees, the information flow among them is restricted. This results in inadequate representation of critical device usability considerations in the decision-making process. While all participants view device safety as an important consideration in the selection process, administrators (who are the final decision-makers) tend to equate safety with technical accuracy and reliability, paying less attention to the role of human factors in safe device use. Findings suggest that collaborative communication technology and automated evidence-based guidelines could provide support to institutional decision-making, ensuring that the process is efficient, effective, and ultimately safe for the patients. PMID- 14552846 TI - Usability in the real world: assessing medical information technologies in patients' homes. AB - OBJECTIVE: This paper presents an approach to usability evaluation of computer based health care systems designed for patient use in their homes. Although such devices are becoming more prevalent, there is very little known about their usability. DESIGN: The theoretical foundations for the methods are discussed. The approach incorporates a cognitive walkthrough usability evaluation and new methods for usability testing that can be conducted in patient's homes. The method was applied to the IDEATel intervention, a multi-institution randomized controlled trial of the feasibility, acceptability, and clinical utility of a home-based telemedicine system for diabetic Medicare population. The usability study was designed to assess barriers to optimal use of the system. The focus was both on dimensions of the interface and on dimensions of patient skills and competency. The usability field research involved testing 25 patients in their homes using the system. The analysis included a range of video-analytic methods of varying levels of granularity. RESULTS: The usability evaluation revealed aspects of the interface that were sub-optimal and impeded the performance of certain tasks. It also found a range of patient-related factors such as numeracy and psychomotor skills that constituted barriers to productive use. CONCLUSIONS: A multifaceted usability approach provided important insight regarding use of technology by an elderly chronic-care patient population and more generally, for understanding how home health initiatives can more effectively use such technology. PMID- 14552847 TI - Medical errors as a result of specialization. AB - Errors in medicine result in over 44,000 preventable deaths annually. Some of these errors are made by specialized physicians at the time of diagnosis. Building on error frameworks proposed in the literature, we tested the experimental hypothesis that physicians within a given specialty have a bias in diagnosing cases outside their own domain as being within that domain. Thirty-two board-certified physicians from four internal medicine subspecialties worked four patient cases each. Verbal protocol analysis and general linear modeling of the numerical data seem to confirm the experimental hypothesis, indicating that specialists try to "pull" cases toward their specialty. Specialists generate more diagnostic hypotheses within their domain than outside, and assign higher probabilities to diagnoses within that domain. PMID- 14552848 TI - Effective drug-allergy checking: methodological and operational issues. AB - Adverse drug events cause a large number of injuries, and adverse events caused by medications administered in the face of known allergies represent an important preventable cause of patient harm. Computerized systems can effectively prevent reactions due to known allergies, but building an effective allergy prevention feature is challenging and presents many interesting informatics issues that have both methodological and operational implications. In this paper, we present the experiences from one large delivery system in delivering allergy-related decision support, discuss some of the different approaches that we have used, and then propose a future approach. We also discuss the methodological, behavioral, and operational issues that have arisen which have a major impact on success. Key factors in drug-allergy checking include storing patient allergy data in a single common repository, representing allergy data using suitable terminologies and creating groups of allergies for inferencing purposes, being judicious about which allergy warnings to display, conveying the reaction that the patient has experienced when exposed to the drug to inform the provider of the importance of the warning, and perhaps most important, implementing strategies to optimize the likelihood that allergy information will be entered. PMID- 14552849 TI - Design and implementation of a comprehensive outpatient Results Manager. AB - Prior research has demonstrated that clinicians often fail to review and act upon outpatient test results in a timely and appropriate manner. To address this patient safety and quality of care issue, Partners Healthcare has developed a browser-based, provider-centric, comprehensive results management application to help clinic physicians review and act upon test results in a safe, reliable, and efficient manner. The application, called the Results Manager, incorporates extensive decision support features to classify the degree of abnormality for each result, presents guidelines to help clinicians manage abnormal results, allows clinicians to generate result letters to patients with predefined, context sensitive templates and prompts physicians to set reminders for future testing. In this paper, we outline the design process and functionality of Results Manager. We also discuss its underlying architectural design, which revolves around a clinical event monitor and a rules engine, and the methodological challenges encountered in designing this application. PMID- 14552850 TI - A tiered approach is more cost effective than traditional pharmacist-based review for classifying computer-detected signals as adverse drug events. AB - OBJECTIVE: To develop a cost-efficient method for identifying adverse drug events (ADEs) and medication errors (MEs) identified using outpatient electronic medical records within ambulatory settings. DESIGN: Comparison of sensitivity and cost of "traditional" pharmacist based approach to identifying ADEs and MEs during a 4 month period with a tiered approach. RESULTS: The proportion of computer generated signals analyzed identified as ADEs were similar using the two approaches while the number of MEs was nearly double with tiered reviews suggesting the same or better sensitivity. Traditional pharmacist review cost $68.70 US dollars to detect an ADE and tiered approach cost only $42.40. CONCLUSION: Tiered review of ADEs and MEs by personnel with increasing clinical capability is more cost-efficient than pharmacist review. PMID- 14552851 TI - Assessing explicit error reporting in the narrative electronic medical record using keyword searching. AB - BACKGROUND: Many types of medical errors occur in and outside of hospitals, some of which have very serious consequences and increase cost. Identifying errors is a critical step for managing and preventing them. In this study, we assessed the explicit reporting of medical errors in the electronic record. METHOD: We used five search terms "mistake," "error," "incorrect," "inadvertent," and "iatrogenic" to survey several sets of narrative reports including discharge summaries, sign-out notes, and outpatient notes from 1991 to 2000. We manually reviewed all the positive cases and identified them based on the reporting of physicians. RESULT: We identified 222 explicitly reported medical errors. The positive predictive value varied with different keywords. In general, the positive predictive value for each keyword was low, ranging from 3.4 to 24.4%. Therapeutic-related errors were the most common reported errors and these reported therapeutic-related errors were mainly medication errors. CONCLUSION: Keyword searches combined with manual review indicated some medical errors that were reported in medical records. It had a low sensitivity and a moderate positive predictive value, which varied by search term. Physicians were most likely to record errors in the Hospital Course and History of Present Illness sections of discharge summaries. The reported errors in medical records covered a broad range and were related to several types of care providers as well as non health care professionals. PMID- 14552852 TI - Retrospective data collection and analytical techniques for patient safety studies. AB - To enhance patient safety, data about actual clinical events must be collected and scrutinized. This paper has two purposes. First, it provides an overview of some of the methods available to collect and analyze retrospective data about medical errors, near misses, and other relevant patient safety events. Second, it introduces a methodological approach that focuses on non-routine events (NRE), defined as all events that deviate from optimal clinical care. In intermittent in person surveys of anesthesia providers, 75 of 277 (27%) recently completed anesthetic cases contained a non-routine event (98 total NRE). Forty-six of the cases (17%) had patient impact while only 20 (7%) led to patient injury. In contrast, in the same hospitals over a two-year period, we collected event data on 135 cases identified with traditional quality improvement processes (event incidence of 0.7-2.7%). In these quality improvement cases, 120 (89%) had patient impact and 74 (55%) led to patient injury. Preliminary analyses not only illustrate some of the analytical methods applicable to safety data but also provide insight into the potential value of the non-routine event approach for the early detection of risks to patient safety before serious patient harm occurs. PMID- 14552853 TI - Mining complex clinical data for patient safety research: a framework for event discovery. AB - Successfully addressing patient safety requires detecting medical events effectively. Given the volume of patients seen at medical centers, detecting events automatically from data that are already available electronically would greatly facilitate patient safety work. We have created a framework for electronic detection. Key steps include: selecting target events, assessing what information is available electronically, transforming raw data such as narrative notes into a coded format, querying the transformed data, verifying the accuracy of event detection, characterizing the events using systems and cognitive approaches, and using what is learned to improve detection. PMID- 14552854 TI - Detecting adverse events for patient safety research: a review of current methodologies. AB - Promoting patient safety is a national priority. To evaluate interventions for reducing medical errors and adverse event, effective methods for detecting such events are required. This paper reviews the current methodologies for detection of adverse events and discusses their relative advantages and limitations. It also presents a cognitive framework for error monitoring and detection. While manual chart review has been considered the "gold-standard" for identifying adverse events in many patient safety studies, this methodology is expensive and imperfect. Investigators have developed or are currently evaluating, several electronic methods that can detect adverse events using coded data, free-text clinical narratives, or a combination of techniques. Advances in these systems will greatly facilitate our ability to monitor adverse events and promote patient safety research. But these systems will perform optimally only if we improve our understanding of the fundamental nature of errors and the ways in which the human mind can naturally, but erroneously, contribute to the problems that we observe. PMID- 14552856 TI - Haemonchus contortus: molecular characterisation of a small heat shock protein. AB - A cDNA encoding a predicted small heat shock protein, HSP20, was isolated from the parasitic nematode Haemonchus contortus. This cDNA encoded a predicted protein of 156 amino acids, which had high sequence identity with other nematode small heat shock proteins. Southern blot analysis of genomic DNA suggested that in H. contortus HSP20 is encoded by a single copy gene. The HSP20 transcript and protein were expressed in the infective larvae (L3), early L4 and adult stages, but expression was not increased by heat shock treatment. In situ hybridisation analysis was used to localise expression of HSP20 mRNA in the adult parasite. Similar HSPs (heat shock protein) were detected by Western blotting in Ancylostoma caninum, Dictyocaulus viviparus, and Toxocara canis, but not in Trichostronglyus colubriformis. The conservation of HSP20 in several different nematode species may reflect its importance to parasites that require mammalian hosts as a part of their development. Index Descriptors and Abbreviations: Haemonchus contortus; nematode; small heat shock protein; L3, infective larvae; xL3, exsheathed L3; eL4, early L4; EST, expressed sequence tag; HSP20, heat shock protein 20; sHSP, small heat shock protein PMID- 14552855 TI - Nitric oxide synthase in Entamoeba histolytica: its effect on rat aortic rings. AB - NADPH-diaphorase activity has been considered as a nitric oxide synthase (NOS) marker. Therefore, the presence of NADPH-d activity in Entamoeba histolytica suggests that they have NOS activity. The aim of this work was to provide support for this contention. The amebic culture medium or amebic purified proteins induced relaxation of endothelium-denuded rat aortic rings pre-contracted with phenylephrine (10(-6) M), which was inhibited when the amebas were incubated with NG-monomethyl-L-arginine or aminoguanidine (NOS inhibitors), or by pretreatment of the aortic rings with methylene blue. L-Arginine reverted the L-NAME inhibitory effect. In addition, trophozoites produce NO in culture and they have proteins which were recognized by antibodies specific to NOS and show activity of NO synthase. In conclusion, our results provide evidence about the production of NO by trophozoites. This molecule may be responsible for the relaxation elicited by the amebic culture medium and may participate in the pathogenesis of the invasive amebiasis. Index Descriptors and Abbreviations: Entamoeba histolytica; NO, nitric oxide; NOS, nitric oxide synthase; iNOS, inducible nitric oxide synthase; ecNOS, endothelial nitric oxide synthase; NADPH-d, NADPH-diaphorase enzyme; beta-NADPH, beta-nicotinamide-adenine dinucleotide; L-NAME, N-omega-nitro L-arginine methyl ester hydrochloride; NBT, nitobluetetrazolium; PBS, phosphate buffered saline; EDTA, ethylenediaminetetraacetic acid; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis PMID- 14552857 TI - Leishmania (Leishmania) amazonensis: differential expression of proteinases and cell-surface polypeptides in avirulent and virulent promastigotes. AB - A comparative study of proteolytic enzymes and cell-surface protein composition in virulent and avirulent Leishmania (Leishmania) amazonensis promastigote forms was carried out using one- and two-dimensional dodecyl sulfate sodium polyacrylamide gel electrophoresis (SDS-PAGE). The surface iodinated protein profiles showed two major polypeptides of 65-60 and 50-47 kDa that were expressed in both virulent and avirulent promastigote forms. However, minor quantitative differences were observed in the cell-surface profile between the avirulent and virulent promastigotes. These included polypeptides of 115, 52, 45, 32, and 25 kDa that were preferentially expressed in the virulent forms. Two-dimensional SDS PAGE showed an accentuated expression of acidic polypeptides; some of them differentially expressed in the promastigote forms analyzed. Live parasites treated with glycosylphosphatidylinositol (GPI)-specific phospholipase C (PLC) from Trypanosoma brucei and immunoprecipitated with the cross-reacting determinant (CRD) antibody recognized three major polypeptides of 65-60, 52, and 50-47 kDa, hence suggesting that these peptides were anchored to the plasma membrane domains through GPI anchor. Moreover, the polypeptides of 65-60 and 52 kDa were also recognized by the gp63 antiserum. Several metalloproteinase activities were similar in both virulent and avirulent promastigote forms, whereas cysteine proteinase activities, sensitive to E-64, were preferentially expressed in virulent promastigotes. These results suggest that cell-surface polypeptides and intracellular cysteine proteinases might play an important role in the virulence of L. (L.) amazonensis. PMID- 14552858 TI - Ascaris suum: cDNA microarray analysis of 4th stage larvae (L4) during self-cure from the intestine. AB - There is spontaneous cure of a large portion of Ascaris suum 4th-stage larvae (L4) from the jejunum of infected pigs between 14 and 21 days after inoculation (DAI). Those L4 that remain in the jejunum continue to develop while those that have moved to the ileum are eventually expelled from the intestines. Although increases in intestinal mucosal mast cells and changes in local host immunity are coincidental with spontaneous cure, the population of L4 that continue to develop in the jejunum may counteract host protective mechanisms by the differential production of factors related to parasitism. To this end, a cDNA library was constructed from L4 isolated from pig jejunum at 21 DAI, and 93% of 1920 original clones containing a single amplicon in the range 400-1500 bp were verified by gel electrophoresis and printed onto glass slides for microarray analysis. Fluorescent probes were prepared from total RNA isolated from: (1) 3rd stage larvae from lung at 7 DAI, (L3); (2) L4 from jejunum at 14 DAI (L4-14-J); (3) L4 from jejunum at 21 DAI (L4-21-J); (4) L4 from ileum at 21 DAI (L4-21-I, and; (5) adults (L5). Cy3-labeled L3, L4-14-J, L4-21-I and L5 cDNA, and Cy5-labeled L4-21 J cDNA were simultaneously used to screen the printed arrays containing the L4-21 J-derived cDNA library. Several clones showed consistent differential gene expression over two separate experiments and were grouped into 3 distinct transcription patterns. The data showed that sequences from muscle actin and myosin, ribosomal protein L11, glyceraldehyde-3-phosphate dehydrogenase and the flavoprotein subunit of succinate dehydrogenase were highly expressed in L4-21-J, but not in L4-21-I; as were a collection of un-annotated genes derived from a worm body wall-hypodermis library, and a testes germinal zone tissue library. These results suggest that only actively developing A. suum L4 are destined to parasitize the host and successfully neutralize host protective responses. PMID- 14552859 TI - Trypanosoma cruzi: partial characterization of minor cruzipain isoforms non adsorbed to Concanavalin A-Sepharose. AB - The present paper reports the partial characterization of a subset of atypical cruzipain molecules which do not bind to Concanavalin A-Sepharose column. They are present in different strains of epimastigote forms of Trypanosoma cruzi and represent a 2-4% of total cruzipain. They were purified by affinity chromatography on Cystatin-Sepharose, recognized by the polyclonal anti-cruzipain serum, and their activity in gelatin-containing gels was completely abolished by E-64, TLCK, leupeptin, and aprotinin but not by PMSF, pepstatin A, EDTA or 1,10 phenantroline. These cysteine proteinases, as well as cruzipain showed to be endoproteinases able to hydrolize azocasein, hemoglobin, and bovine serum albumin at acidic pHs. However, evidences are presented indicating that this subset of cruzipain isoforms were also able to use the same blocked chromogenic peptidyl substrates than cruzipain at similar optimal alkaline pH values although with a different order of preference. Moreover, they showed a different oligosaccharide pattern after enzymatic treatment by high pH anion exchange chromatography, suggesting that this structural difference may account for the atypical behaviour in the lectin column. PMID- 14552860 TI - Non-classic characteristics define prominent DNase activities from the intestine and other tissues of Haemonchus contortus. AB - The anthelmintic fenbendazole (FBZ) induces nuclear DNA fragmentation (DF) in intestinal cells of Haemonchus contortus. The DNA fragments had 3'-OH, which suggests involvement of a neutral DNase. To identify candidate DNase(s) involved, DNase activity in H. contortus intestine and other worm fractions was characterized relative to classic DNases I (neutral) and II (acidic). Seven distinct DNase activities were identified and had Mrs of 34, 36, 37 or 38.5 kDa on zymographic analysis. The different activities were distinguished according to pH requirement, sensitivity to 10 mM EDTA and worm compartment. Activities of intestinal DNases at 34, 36 and 38.5 kDa were sensitive to EDTA at pH 5.0 and 7.0. Sensitivity to EDTA at pH 5.0 was unexpected compared to classic acidic DNase II activity, suggesting unusual properties of these DNases. In whole worms, however, the activities at 36 and 38.5 kDa were relatively insensitive to EDTA, indicating predominance of DNases that are distinct from the intestine. The activity at 37 kDa in excretory/secretory products had an acidic pH requirement and was insensitive to EDTA, resembling classic acidic DNase activity. Under conditions of pH 5.0 and 7.0, intestinal DNases produced 3'-ends that could be labeled by terminal deoxynucleotidyl transferase, indicating presence of 3'-OH. The labeling of 3'-ends at pH 5.0, again, was unexpected for acidic DNase activity. These results and several other activities suggest that multiple H. contortus DNases have characteristics distinct from the classic mammalian DNases I and II. Treatment of H. contortus with FBZ did not induce any detectable DNase activities distinct from normal intestine, although relative activities of intestinal DNases appear to have been altered by this treatment. PMID- 14552861 TI - Searching for promoter activity in RIME/Ingi retrotransposons from Trypanosoma brucei: binding of a nuclear protein to their 5' extremity. AB - In Trypanosoma brucei only two promoters for protein-encoding genes have been characterized so far. The RIME and Ingi elements of T. brucei are similar in structure to the non-long terminal repeat retrotransposons. Internal promoters usually located at their 5' end drive transcription of several of the latter elements. During a search for promoter activity within RIME and Ingi we focused on a region at the 5' end of both elements, which we termed rime5. A 50 kDa nuclear protein was found to specifically bind to the double strand and single strand sense of rime5 DNA. However, constructs containing several rime5 fragments inserted upstream of a chloramphenicol acetyltransferase (CAT) reporter gene failed to promote both transcription and expression of this gene in transient transfection assays. Finally, we have analyzed the expression of the Ingi elements and despite the high level of transcripts detectable in the cytoplasm, antibodies raised against two different domains of the single open reading frame did not detect any component in total extracts from T. brucei, suggesting that few Ingi copies, if any, are actually active. PMID- 14552863 TI - Cruzella marina (Bodonina, Kinetoplastida): non-catenated structure of poly kinetoplast DNA. PMID- 14552862 TI - Neospora caninum and Toxoplasma gondii: a novel adhesion/invasion assay reveals distinct differences in tachyzoite-host cell interactions. AB - This paper describes an adhesion/invasion assay, based on combined pyrrolidine dithiocarbamate (PDTC) and antibody treatment of parasites followed by quantitative real-time PCR. This PDTC-PCR assay can be used to comparatively assess the participation of host cell- and parasite-associated components during host cell adhesion and entry by Neospora caninum and Toxoplasma gondii tachyzoites, respectively, and is potentially applicable to any other apicomplexan parasite. The assay allows to determine the parasite invasion rate in relation to the overall number of parasites which interact with host cells in any given experiment, and thus represents a significant improvement to conventional microscopic assays in terms of accuracy and reproducibility. Using this assay it was possible to show that adhesion and invasion of N. caninum tachyzoites are two distinct and separated events, in that N. caninum tachyzoites preferentially utilise host cell surface chondroitin sulphates for adhesion, but not for the host cell invasion process. Application of the PDTC-PCR assay also demonstrated that N. caninum and T. gondii tachyzoites differ largely with regard to the functional involvement of proteases in adhesion and invasion of host cells. Thus, although phylogenetically closely related, N. caninum and T. gondii are biologically quite different and exhibit distinct dissimilarities with regard to host cell interactions. PMID- 14552864 TI - Genetic characterization by random amplified polymorphic DNA analysis (RAPD) of 18 isolates of Giardia lamblia obtained from day care children. PMID- 14552865 TI - Tales of transdifferentiation. PMID- 14552866 TI - Primate models of Parkinson's disease. PMID- 14552867 TI - The p38 MAP kinase signaling pathway in Alzheimer's disease. PMID- 14552868 TI - Novel crosstalk between G protein-coupled receptors and NMDA receptors. PMID- 14552869 TI - alpha6beta1 integrin directs migration of neuronal precursors in adult mouse forebrain. AB - New neuroblasts are constantly generated in the adult mammalian subventricular zone (SVZ) and migrate via the very-restricted rostral migratory stream (RMS) to the olfactory bulb, where they differentiate into functional neurons. Several facilitating and repulsive molecules for this migration have been identified, but little is known about chemoattractive molecules involved in the directed nature of this migration in vivo. Here, we investigated the role of the alpha6beta1 integrin, and its ligand, laminin, in controlling guidance of the migrating neuroblasts in adult mice. Immunostaining for the alpha6beta1 integrin was present in neuroblasts and their processes in the anterior/rostral SVZ and the RMS. Inhibition of the endogenous alpha6 or beta1 subunit with locally injected antibodies disrupted the cohesive nature of the RMS, but did not kill the neuroblasts. Infusion of a 15 a.a. peptide, representing the E8 domain of the laminin alpha chains that bind alpha6beta1 integrin, into the neostriatum redirected the neuroblasts away from the RMS towards the site of infusion. Injection of a narrow tract of intact laminin also drew the neuroblasts away from the RMS, but in a more restricted localization. These results suggest a critical role for integrins and laminins in adult SVZ-derived neuroblast migration. They also suggest that integrin-based strategies could be used to direct or restrict neuroblasts to CNS regions where they are needed for cell replacement therapies in the nervous system. PMID- 14552870 TI - Expression of the type 1 and type 2 receptors for tumor necrosis factor after traumatic spinal cord injury in adult rats. AB - Posttraumatic inflammation has been implicated in secondary tissue damage after spinal cord injury (SCI). Tumor necrosis factor-alpha (TNF-alpha) is a key inflammatory mediator that is increasingly expressed after SCI. The effect of TNF alpha is mediated through its receptors TNFR1 (p55) and TNFR2 (p75). However, whether these two receptors are expressed after SCI has not been demonstrated. In the present study, the temporo-spatial expression of TNFR1 and TNFR2 was examined in rats that had received a 10 g impact injury dropped at a height of 12.5 mm using the New York University impact device. In sham operates, no detectable TNFR1 or TNFR2 immunoreactivity (IR) was observed. In contused spinal cord, TNFR1 protein expression and immunoreactivity (IR) were detected as early as 15 min postinjury, reached its peak at 8 h, and declined markedly after 1 and 3 days postinjury. The temporal pattern of TNFR2 expression was similar to that of TNFR1 but its expression peaked at 4 h postinjury. During peak expression, TNFR1- and TNFR2-IR were most intense at the site of injury and decreased gradually from the injury epicenter. TNFR1- and TNFR2-positive cells included neurons, astrocytes, and oligodendrocytes. Methylprednisolone (MP), a synthetic glucocorticoid, partially inhibited the injury-induced expression of TNFR1 and TNFR2, an effect which could be reversed by RU486, an antagonist of glucocorticoid receptors. We suggest that the expression of TNFR1 and TNFR2 after SCI may contribute to posttraumatic inflammatory responses of TNF-alpha. PMID- 14552871 TI - Endogenous and exogenous ciliary neurotrophic factor enhances forebrain neurogenesis in adult mice. AB - Neurogenesis in the adult mammalian CNS occurs in the subventricular zone (SVZ) and dentate gyrus. The receptor for ciliary neurotrophic factor (CNTF), CNTFRalpha, is expressed in the adult subventricular zone. Because the in vitro effects of CNTF on neural precursors have been varied, including proliferation and differentiation into neurons or glia, we investigated its role in vivo. Injection of CNTF in the adult C57BL/6 mice forebrain increased the number of cells labeled with ip BrdU in both neurogenic regions. In the dentate gyrus, CNTF also appeared to enhance differentiation of precursors into neurons, i.e., increased the proportion of NeuN+/BrdU+ cells from approximately 14 to approximately 29%, but did not affect differentiation into astrocytes (GFAP+) or oligodendrocytes (CNPase+). In the SVZ, CNTF increased the proportion of GFAP+/BrdU+ cells from approximately 1 to approximately 2%. CNTF enhanced the distance of migration of new neurons into the granule cell layer. Intraventricular injection of neutralizing anti-CNTF antibodies reduced the number of BrdU-labeled cells in the SVZ. These results suggest that endogenous CNTF regulates adult neurogenesis by increasing proliferation of neural stem cells and/or precursors. Alternatively, CNTF could maintain cells longer in the S phase, resulting in increased BrdU labeling. In the neurogenic region of the SVZ, CNTFRalpha was exclusively present in GFAP-positive process-bearing cells, suggesting that CNTF affects neurogenesis indirectly via neighboring astroglia. Alternatively, these cells may be part of the neural precursor lineage. The restricted expression of CNTF within the nervous system makes it a potential selective drug target for cell replacement strategies. PMID- 14552872 TI - Enkephalin, neurotensin, and substance P immunoreactivite neurones of the rat GP following 6-hydroxydopamine lesion of the substantia nigra. AB - The ascending dopaminergic tract influences the activity of GP neurones in normal conditions. Its lesion may lead to an up-regulation of activity in this nucleus that is contrary to what would be expected based on the current model of the basal ganglia function. In this study we investigated the occurrence of enkephalin, neurotensin, and substance P immunoreactivity of the rat globus pallidus (GP) following lesion of the nigrostriatal pathway induced by the injection of the toxin 6-hydroxydopamine into the substantia nigra. Since 60-65% of GP neurones are immunopositive for parvalbumin, the immunoreactivity for peptides was evaluated, considering the different content in parvalbumin of pallidal neurones types, at early and chronic phases of denervation. Our results showed that a lesion of the nigrostriatal pathway induced the expression of enkephalin, neurotensin, and substance P immunoreactivity in numerous pallidal cell bodies. Each subgroup of neurones showed a different pattern of distribution. These modifications equally involved the two main subclasses of neurones. However parvalbumin-negative neurones were modified to a larger extent than the parvalbumin-positive ones. These data indicate that nigrostriatal lesion induces in a wide and unexpected peptide synthesis at least in three different subgroups of GP neurones. These modifications might be useful to further histochemically characterise neurones of the GP. PMID- 14552873 TI - Transplantation of neural stem cells modulates apolipoprotein E expression in a rat model of stroke. AB - The expression of apolipoprotein E (apoE) after ischemic brain damage has been associated with plasticity involved in promoting functional recovery. We therefore examined the expression and distribution of apoE in rats that received intraparenchymal grafts of the conditionally immortal stem cell line MHP36 either ipsilateral or contralateral to the lesion or intraventricular grafts 4 months after transplantation. ApoE immunoreactivity was highly expressed in the striatum, somatosensory cortex, and thalamus of the lesioned hemisphere in all rats subjected to middle cerebral artery occlusion. Only in rats with intraparenchymal grafts, apoE was significantly upregulated in the contralateral hemisphere, whereas levels and distribution in rats with intraventricular grafts resembled those of ischemic controls. In ischemic rats, apoE was seen in both astrocytes and neurons on the lesioned side, and in grafted rats, apoE was present in host and transplanted neurons and astrocytes. Previously we have shown that intraparenchymal grafts reduced sensorimotor asymmetry, whereas intraventricular grafts improved cognitive dysfunction, with transplanted cells being widely distributed in cortex, striatum, and corpus callosum on both sides of the brain in all grafted groups. Thus, stem cells grafted in the parenchyma are not only capable of limited expression of apoE in the host brain but also trigger a robust increase on the side contralateral to stroke damage where this does not normally occur. Findings that parenchymal, but not ventricular, grafts facilitated sensorimotor recovery suggests that apoE might contribute to plastic changes in relevant pathways, possibly on both sides of the brain. In contrast, no evidence was found for an association between apoE and recovery of cognitive function in rats with intraventricular grafts. PMID- 14552874 TI - Taurine and the control of basal hormone release from rat neurohypophysis. AB - Pituicytes of pituitary neural lobe are rich in the amino acid taurine, which they release upon hypoosmotic stimulation. As a generally inhibitory amino acid, taurine is thought to activate receptors on neural lobe nerve terminals and exert some control over hormone release. Previous work has shown the presence of glycine and GABA(A) receptors in neural lobe, both of which have affinity for taurine. Using a perifused explant system, we studied the effects of taurine activation of glycine and GABA(A) receptors on basal hormone release. Somewhat surprisingly, taurine induced increases in basal release of both vasopressin and oxytocin. Taurine-induced increases in oxytocin release were blocked by bicuculline, suggesting involvement of GABA(A) receptors. Increases in vasopressin release were not blocked by bicuculline, indicating involvement of receptors other than GABA(A). Although combined bicuculline and strychnine, an antagonist at most glycine receptors, also did not block increased vasopressin release, picrotoxin (a Cl(-) channel blocker) was effective in blocking increases in both vasopressin and oxytocin release. The other receptor(s) involved in taurine actions is postulated to be strychnine-insensitive glycine receptors. Thus, taurine in neural lobe may act via both a GABA(A) receptor and one or more types of glycine receptors to depolarize nerve terminal membranes under basal conditions. Taurine-induced partial depolarization resulting in Na(+) channel inactivation is probably responsible for its previously observed inhibition of stimulated hormone release from neural lobe. PMID- 14552875 TI - AVP V1a-R expression in the rat hypothalamus around parturition: relevance to antipyresis at term. AB - An endogenous antipyresis has been observed around parturition in several species, including rats. It has been proposed that the neuropeptide vasopressin is responsible for this antipyresis via an action on the V(1a) receptor subtype, but this concept is controversial. We therefore addressed the question of the regulation of V(1a) receptor expression within the rat hypothalamus around parturition, to assess its possible involvement in the antipyresis phenomenon observed at term. We analyzed V(1a) receptor mRNA and protein levels in the hypothalamus/preoptic area of female rats at Days 15 and 22 (parturition) of gestation, and at Day 5 of lactation. We used quantitative RT-PCR to assess the mRNA levels and designed a semiquantitative Western blot assay to analyze changes in protein levels between the three stages studied. No significant changes either in V(1a) receptor mRNA or protein levels were observed between the three stages, suggesting that variations in the hypothalamic V(1a) receptor expression levels alone cannot account for the endogenous antipyresis observed at term. PMID- 14552876 TI - Neuroprotection and neuronal dysfunction upon repetitive inhibition of oxidative phosphorylation. AB - Repetitive inhibition of oxidative phosphorylation is an established model of neurodegeneration. In contrast, a single mild treatment can be neuroprotective chemical preconditioning. Repetitive chemical inhibition of oxidative phosphorylation may thus be a tool to study deterioration and improvement of cellular hypoxic tolerance and subsequent differential regulation of cellular responses in the same model. We investigated murine hippocampal function upon repetitive intraperitoneal injections of 3-nitropropionate (3-NP; 20 mg/kg body weight), an inhibitor of mitochondrial complex II. With a 2-day interval of repetitive in vivo treatment with 3-NP, posthypoxic recovery of population spike amplitude was below control. In contrast, even after nine in vivo treatments with 3-NP at 4-day intervals, an almost complete recovery of population spike amplitude was observed. Nerve growth factor (NGF) as assessed by ELISA and expression of beta-amyloid precursor protein (APP) mRNA increased upon nine treatments at 2-day intervals, but remained at control levels with 4-day intervals. In contrast, brain-derived neurotrophic factor (BDNF) as assessed by ELISA increased with the latter treatment. Expression of mRNA for adenosine-A1 and -A3 receptors and endothelial and neuronal nitric oxide synthase remained at control level for both treatment intervals. We conclude that the time interval between mild, subclinical repetitive inhibition of oxidative phosphorylation determines hippocampal neuronal impairment and integrity and modulates NGF and BDNF differently. Decreased hypoxic tolerance and increased APP expression upon repetitive inhibition of oxidative phosphorylation at short time intervals may thus trigger a vicious cycle and be a cofactor for neuronal dysfunction in cerebral hypoxia and neurodegenerative diseases. PMID- 14552877 TI - Improvement of neurological deficits by intracerebral transplantation of human adipose tissue-derived stromal cells after cerebral ischemia in rats. AB - Mesenchymal stem cells can be expanded rapidly in vitro and differentiated into multiple mesodermal cell types. In addition, their differentiation into neuron like cells expressing markers typical for mature neurons has been reported. We isolated human adipose tissue stromal cells (hATSCs) from human liposuction tissues and induced neural differentiation with azacytidine. Following neural induction, hATSCs changed toward neural morphology and displayed expression of MAP2 and GFAP. hATSCs, which were labeled with LacZ adenovirus, were injected into the lateral ventricle of the rat brain. Transplanted cells migrated to various parts of the brain, and ischemic brain injury by middle cerebral artery occlusion (MCAo) increased their migration to the injured cortex. Some of the transplanted cells expressed MAP2 and GFAP. Transplantation of hATSCs improved functional deficits in ischemic brain injury induced by MCAo. Intracerebral grafting of BDNF-transduced hATSCs significantly improved motor recovery of functional deficits in MCAo rats. These data indicate that transplanted hATSCs survive, migrate, and improve functional recovery after stroke and that genetically engineered hATSCs can express biologically active gene products and, therefore, can function as effective vehicles for therapeutic gene transfer to the brain. PMID- 14552878 TI - Ectopic expression of the TrkA receptor in adult dopaminergic mesencephalic neurons promotes retrograde axonal NGF transport and NGF-dependent neuroprotection. AB - A recombinant adeno-associated virus (rAAV) was used to investigate the impact of an ectopic expression of the NGF high-affinity receptor in adult neurons. The rat TrkA cDNA cloned in a pCMX vector was first tagged with a human c-Myc sequence. The resulting vector was shown to encode a functional receptor which promoted the expression of TrkA immunoreactivity upon transfection of 293 fibroblasts or nnr5 cells, a TrkA-defective variant of PC12 cells. These cells also accumulate TrkA transcripts upon transfection and extended neurites in the presence of NGF. Therefore, the TrkA(myc) cassette was inserted into the pSSV9 plasmid. The new vectors shared properties similar to pCMX TrkA(myc) in 293 and nnr5 cells and enabled the preparation of rAAV TrkA(myc) viruses. Unilateral injection of this rAAV into the substantia nigra (SN) resulted in a protracted expression of TrkA (or c-Myc) immunoreactivity in numerous cell bodies, including tyrosine hydroxylase (TH)-positive dopaminergic neurons. The presence of TrkA receptors in corresponding striatal dopaminergic endings was demonstrated by the advent of a striato-nigral retrograde axonal transport of (125)I-NGF. Likewise, ectopic expression of TrkA in neurons of the parafascicular thalamic nucleus promoted a striatofuge transport of NGF toward this structure. To investigate whether ectopic expression of TrkA in SN neurons may confer neuroprotection, lesions were induced by 6-hydroxydopamine in striata located ipsilateral to the virus injection site. NGF or vehicle were next delivered dorsally to the virus-treated SN for 2 weeks, before sacrifice and processing of brains for TH immunohistochemistry. NGF treatment, in contrast to treatment with vehicle, significantly enhanced the number of dopaminergic neurons counted in the lesioned SN. These data suggest that ectopic TrkA can mediate the trophic actions of NGF and influence neuronal plasticity in vivo. PMID- 14552879 TI - Glial cell line-derived neurotrophic factor-enriched bridging transplants promote propriospinal axonal regeneration and enhance myelination after spinal cord injury. AB - Glial cell line-derived neurotrophic factor (GDNF), a distant member of the transforming growth factor-beta (TGF-beta) family, is widely expressed in the developing and adult central nervous system (CNS). At present, limited information is available regarding the effects of GDNF in the repair of spinal cord injury (SCI). In the present study, mini-guidance channels containing either: (1) Matrigel (MG, a basement membrane component), (2) Schwann cells (SCs, 120 x 10(6)/ml) in MG (SC-MG), (3) recombinant human GDNF (rhGDNF, 3 microg/microl) in MG (GDNF-MG), and (4) a combination of all three components (GDNF-SC-MG) were grafted into a T9 hemisection-gap lesion in adult rats to examine the effects of GDNF on axonal regeneration and myelination following SCI. Thirty days post-transplantation, limited axonal growth was observed within guidance channels containing MG-alone (MG). When SCs were added to the channels (SC-MG group), consistent axonal ingrowth containing both myelinated and unmyelinated axons was observed, confirming our previous findings. The addition of GDNF-alone without SCs (GDNF-MG) resulted in substantial ingrowth of unmyelinated axons, suggesting that GDNF has a direct neurite-growth promoting effect on these axons. Implantation of channels containing both GDNF and SCs (GDNF-SC-MG) produced a significant and synergistic increase in axonal regeneration and myelination. In addition, GDNF reduced the extent of reactive gliosis, infiltration of activated macrophages/microglia, and cystic cavitation at the graft-host interfaces. Retrograde tracing revealed that grafts of SC seeded channels containing GDNF promoted a significant increase in the number of propriospinal neurons which had regenerated their axons into the grafts, as compared to SC-MG-seeded channels. These results indicate that GDNF may play a novel therapeutic role in promoting propriospinal axonal regeneration, enhancing myelin formation, and improving graft-host interfaces after SCI. PMID- 14552880 TI - P38 MAP kinase is activated at early stages in Alzheimer's disease brain. AB - The regional, cellular, and subcellular localization of phosphorylated p38 MAPK (pp38) was examined by immunocytochemistry, immuofluorescent multiple labeling, and immunoblotting of extracts as well as immunoprecipitates of human postmortem tissue from control and Alzheimer's disease (AD) cases at different Braak stages. "Early AD" cases (Braak stages IV-V) and a subset of Braak stage VI cases have high levels of pp38 immunoreactivity, with the most dense immunoreactivity located in CA2 and subiculum followed by CA1 in the hippocampus. On the contrary, very little pp38 was detected in age-matched controls (Braak stages 0-II). More importantly, as revealed by various multiple labeling experiments, pp38 immunoreactivity is mainly located in neurons bearing early neurofibrillary pathology, but not in typically fibrillar tangles that are densely stained by thioflavin-S. Most pp38-positive neurons only contain a small amount of phospho tau. Additionally, pp38 immunoreactivity was not associated with senile plaques. At the subcellular level, pp38-immunoreactive granules are usually larger than the granules stained with the lysosomal marker cathepsin D. Immunoblotting with different extraction buffers and immunoprecipitation indicate that pp38 does not or only loosely binds to phospho-tau. Taken together, this study demonstrates that p38 MAPK is activated at early stages of neurofibrillary degeneration in AD hippocampus. The p38 activation may also be linked to neurodegeneration through mechanisms other than neurofibrillary tangle formation. PMID- 14552882 TI - Sensorimotor deficits in a unilateral intrastriatal 6-OHDA partial lesion model of Parkinson's disease in marmoset monkeys. AB - Animal studies investigating the efficacy of neurotrophic factors as treatments for Parkinson's disease (PD) ideally require partial dopamine (DA) lesion models. The intrastriatal 6-hydroxydopamine (6-OHDA) lesion model may be suitable for this purpose. Although this model has been well characterized in rodents, it has not previously been used in monkeys. The goal of the present study was to characterize the behavioral effects of unilateral injections of 6-OHDA in the basal ganglia of common marmoset monkeys (Callithrix jacchus). Cell counts from tyrosine hydroxylase immunochemistry 5 months postlesion revealed DA cell loss in the substantia nigra on the lesioned side to approximately 46% of relative to the unlesioned side. 6-OHDA lesioned monkeys showed a variety of behavioral deficits. Apomorphine induced rotation and simple sensorimotor measures (head position bias and PD disability rating score) were most affected by the lesion. The largest deficits were seen at 1 or 2 weeks postsurgery but had recovered by week 10. 6 OHDA lesioned monkeys took longer to complete a more complex sensorimotor staircase task. At 3.5 months postlesion, 6-OHDA monkeys also showed deficits on an object retrieval task designed to measure sensorimotor planning and skilled hand use. alpha-Methyl-p-tyrosine, a tyrosine hydroxylase inhibitor, reinstated those deficits which had undergone recovery in the lesioned animals and also exacerbated the deficits on the staircase task. This model has potential in assessing treatments for PD aimed at curtailing disease progression such as continuous delivery of neurotrophic factors. PMID- 14552881 TI - Proliferation and neuronal differentiation of mitotically active cells following traumatic brain injury. AB - Recent studies have identified endogenous neural stem cells in adult rodent brains. The present study characterizes the early response of mitotically active cells in the brain to traumatic brain injury. Animals were subjected to lateral fluid percussion injury and sacrificed at various times after injury. To examine increases in cell proliferation animals were injected with the mitotic marker bromodeoxyuridine (BrdU) 24 h before sacrifice. Increased numbers of mitotically active cells were observed at 2 days in the subgranular zone (SGZ) and the subependymal zone (SEZ) under the injury site. To characterize the differentiation potential of these cells, animals were injected with BrdU 18 and 20 h after injury, then sacrificed at multiple time points after injury. Histologically, co-localization with betaIII-tubulin (neuronal marker) and BrdU was evident at 10 and 15 days postinjury in the SGZ. Flow cytometry analysis was used to quantitatively assess neurogenesis in the SEZ. Animals were sacrificed 1, 5, or 10 days after injury and tissue sections extracted, grown in tissue culture for 24 h, fixed, and stained for nestin and betaIII-tubulin to identify newly formed neurons. The percentage of cells expressing both markers was determined using flow cytometry analysis. There was a significant increase in newly differentiated neurons by 10 days postinjury in the SEZ. Thus, we conclude that traumatic brain injury stimulates an increase in proliferation of endogenous neural stem/progenitor cells and that a significant number of these express a neuronal marker. This response may be the brain's way of trying to heal itself after injury. PMID- 14552883 TI - Cyclosporin A and Bcl-2 do not inhibit quinolinic acid-induced striatal excitotoxicity in rodents. AB - Mitochondrial permeability transition (MPT) is a nonselective inner membrane permeabilization that contributes to neuronal cell death under circumstances such as brain trauma, ischemia, and hypoglycemia. Here we study the participation of MPT and the Bcl-2-sensitive apoptotic cell death pathway in glutamate receptor mediated excitotoxicity. Intrastriatal infusions of the N-methyl-D-aspartate (NMDA) receptor agonist quinolinic acid caused massive striatal neurodegeneration in both rats and mice. Interestingly, transgenic mice overexpressing human Bcl-2 and rats systemically treated with cyclosporin A did not exhibit reduced sensitivity to quinolinic acid-induced striatal toxicity. Both Bcl-2 and cyclosporin A are inhibitors of MPT; in addition Bcl-2 also inhibits apoptotic stimuli-mediated release of mitochondrial apoptogenic factors. Isolated brain mitochondria from cyclosporin A-treated rats showed resistance to Ca(2+)-induced dissipation of the membrane potential, indicating protection against MPT. We conclude that quinolinic acid-mediated striatal excitotoxicity is not dependent on MPT and Bcl-2-sensitive apoptotic cell death pathways. PMID- 14552884 TI - Acute activation of mitogen-activated protein kinases following traumatic brain injury in the rat: implications for posttraumatic cell death. AB - The regional activation (via phosphorylation) of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) signaling pathways was examined using immunoblotting and immunohistochemistry following experimental brain injury. Anesthetized rats were subjected to lateral fluid-percussion brain injury of moderate severity (2.4-2.6 atm) and euthanized at 2, 6, 24, and 72 h after injury; sham-injured animals were surgically prepared but were not injured. Immunohistochemical evidence of activation of JNK and ERK1/2 pathways was observed predominantly in regions that exhibit neural cell apoptosis and axonal damage following brain trauma. Activation of the ERK1/2 pathway was observed as early as 2 h and up to 72 h postinjury in nonneuronal cells in all layers of the cortex at the site of maximal injury, in the white matter below the site of maximal cortical damage and in the thalamus. In contrast, activation of JNK signaling was observed only at 24 and 72 h postinjury in a few neurons at the core of the cortical injury site. However, robust JNK activation was observed between 2 and 72 h postinjury in both axons and nonneuronal cells in the white matter below the site of maximal cortical damage and in the thalamus. Activation of ERK1/2, but not JNK, was observed in cells in the dentate hilus in the hippocampus in both hemispheres between 2 and 24 h postinjury. Immunoblotting analyses of extracts from various brain regions did not reveal significant alterations in intensities of either total or phosphorylated proteins underscoring the focal nature of the immunohistochemical observations. However, these results suggest that activation of MAP kinase signaling pathways may be associated with posttraumatic cell damage and are indicative of the heterogeneous nature of the mechanisms underlying regional cell death following TBI. PMID- 14552885 TI - Induction of astrocytic cytoplasmic phospholipase A2 and neuronal death after intracerebroventricular carrageenan injection, and neuroprotective effects of quinacrine. AB - Glial reaction is often associated with nervous tissue injury, but thus far, few studies have examined whether it can be a cause of neuronal injury. We now study the effect of intracerebroventricular injection of a carrageenan on cytoplasmic phospholipase A(2) (cPLA(2)) expression and neuronal injury in the hippocampus. The enzyme cPLA(2) hydrolyzes neural membrane glycerophospholipids and generates precursors for proinflammatory mediators. An induction of cPLA(2) in astrocytes and death of neurons in the hippocampus were observed following glial reaction induced by intracerebroventricular injections of carrageenan. cPLA(2) levels and neuronal death were modulated by daily intraperitoneal injections of quinacrine, an inhibitor of phospholipase A(2) that can cross the blood brain barrier. These observations support a role for astrocytic cPLA(2) in mediating neuronal death. PMID- 14552886 TI - Increased striatal pre-proenkephalin B expression is associated with dyskinesia in Parkinson's disease. AB - Long-term treatment of Parkinson's disease with levodopa is compromised by the development of motor complications, including on-off fluctuations and involuntary movements termed dyskinesia. The neural mechanisms underlying treatment-related dyskinesias may involve underactivity of the output regions of the basal ganglia, i.e., the medial segment of the globus pallidus (GPm) and substantia nigra pars reticulata (SNR). Increased activity of GABAergic neurons of the "direct" striatopallidal pathway has been implicated in the suppression of the GPm and SNR and thus the development of dyskinesia. The direct pathway uses opioids as a co neurotransmitter. These opioid peptides are products of the high-molecular weight opioid precursor pre-proenkephalin B (PPE-B). In situ hybridisation studies were employed to investigate PPE-B mRNA expression in postmortem striatal tissue from patients with a clinicopathological diagnosis of Parkinson's disease, all of whom displayed levodopa-induced motor complications, including dyskinesia prior to death and in the caudate-putamen (striatum) of the MPTP-lesioned macaque model of Parkinson's disease with treatment-related dyskinesia. Striatal PPE-B mRNA expression was significantly increased by 172% in dyskinetic Parkinson's disease patients compared to age-matched controls. This increase was heterogeneous with increased expression within the striosomes compared to matrix compartments of the striatum. Striatal PPE-B mRNA expression was significantly increased by 185% in the MPTP-lesioned macaque exhibiting dyskinesia, compared to parkinsonian, nondyskinetic MPTP-lesioned macaques, and by 146% compared to non-parkinsonian, nondyskinetic controls. Increased PPE-B mRNA expression, with subsequent elevations in opioid peptide transmission within the direct striatal output pathways, may underlie treatment-related dyskinesia in Parkinson's disease. PMID- 14552888 TI - Estrogen receptor alpha-immunoreactive astrocytes are increased in the hippocampus in Alzheimer's disease. AB - Postmenopausal estrogen use may decrease the risk, and delay the onset and progression, of Alzheimer's disease (AD). By means of fluorescence immunocytochemistry, the present study investigated the distribution of estrogen receptor alpha (ERalpha) in the human hippocampus in controls and in AD cases. ERalpha immunoreactivity was observed in neurons and glial fibrillary acidic protein (GFAP)-immunoreactive astrocytes in the hippocampus both in controls and AD cases. The number and density of GFAP- and ERalpha-positive astrocytes was increased in AD. The number of GFAP-immunoreactive astrocytes, the number of nuclear ERalpha-staining astrocytes, and cytoplasmic ERalpha-staining astrocytes per unit area (1 mm(2)) significantly increased (P < 0.001, P < 0.05, P < 0.05, respectively) in CA1 in AD patients, while the percentage of ERalpha immunoreactive astrocytes of the two groups did not differ (P > 0.05). These data suggest an important role for ERalpha-mediated effects of estrogens on neurons and astrocytes in the hippocampus of human and AD patients. PMID- 14552887 TI - Neurotrophins regulate proliferation and survival of two microglial cell lines in vitro. AB - Microglia are thought to play a key role in the development and regeneration of the central nervous system although the mechanisms regulating their presence and activity are not fully understood. Substantial evidence suggests that members of the neurotrophin family such as nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 and -4 (NT-3/4) have a dramatic effect on both neurons and perineuronal cells. This study employed two murine microglial lines, BV-2 and N9, to examine the action of these neurotrophins on the mitotic activity and survival of microglia in vitro. Neurotrophins were incorporated into the media at the time of plating and cell number and levels of mitochondrial dehydrogenase activity (MTT) were determined at various time points in vitro. NGF increased cell number and MTT levels of both cell lines in a dose dependent manner. BV-2 was more sensitive to NGF than N9. Similar responses were elicited by BDNF, although the sensitivity of each cell line was different than that found for NGF. NT-3 and NT-4 had no effect on cell proliferation. However, NT-4 had an effect on the survival of BV-2 and N9 cells. The response of these cells to neurotrophins was blocked by K252a, a tyrosine kinase inhibitor, suggesting that actions of neurotrophins were mediated by high-affinity tyrosine kinase receptors (Trk). Immunolocalization studies revealed positive Trk (pan) reactivity in the above cell lines and in primary microglia, but an absence of the low-affinity p75 neurotrophin receptor. Western blot analysis supported the above observations. These studies suggest that in addition to their neurotrophic actions, NGF and BDNF may also regulate microglial dynamics, thereby influencing the surrounding milieu during neuronal regeneration. PMID- 14552889 TI - Induction of neuropilins-1 and -2 and their ligands, Sema3A, Sema3F, and VEGF, during Wallerian degeneration in the peripheral nervous system. AB - The neuropilins, NP-1 and NP-2, are coreceptors for Sema3A and Sema3F, respectively, both of which are repulsive axonal guidance molecules. NP-1 and NP 2 are also coreceptors for vascular endothelial growth factor (VEGF). The neuropilins and their ligands are known to play prominent roles in axonal pathfinding, fasciculation, and blood vessel formation during peripheral nervous system (PNS) development. We confirmed a prior report (Exp. Neurol. 172 (2001) 398) that VEGF mRNA levels rise during Wallerian degeneration in the PNS and herein demonstrate that NP-1, NP-2, Sema3A, and Sema3F mRNA levels increase in peripheral nerves distal to a transection or crush injury. In a sciatic nerve crush model, in which axonal regeneration is robust, the highest levels of Sema3F mRNA below the injury site are in the epi- and perineurium. Our results suggest the possibility that the neuropilins and their semaphorin ligands serve to guide, rather than to impede, regenerating axons in the adult PNS. PMID- 14552890 TI - Synemin expression in developing normal and pathological human retina and lens. AB - Synemin (Syn) is an intermediate filament (IF) protein. To gain insight into a morphogenetic role of Syn, we have studied its expression patterns in the developing human retina and lens and compared it with those of other IF proteins. In addition, we have tested Syn expression in fetuses (23 and 28 weeks) affected by Walker-Warburg syndrome (WWS), Meckel syndrome, and trisomy 13. In the retina, Syn expression starts in the nerve fiber and ganglion cell layers (NFL and GCL) at 15 weeks, remains there in up to 20 weeks, and spreads to other layers and may be colocalized with vimentin, GFA, or neurofilaments in the subsequent 16 weeks. This expansion of Synemin expression from 20 to 28 weeks is not observed in WWS in which Syn immunoreactivity in NFL is reduced and Vim expression is increased. Changes are seen in Syn or vimentin expressions in the retinae of 23-week-old Meckel syndrome or 28-week-old trisomy 13 fetuses. Syn expression in the lens is, at first (16 weeks), uniformly distributed, becoming stronger in the epithelium of the anterior part at 25 weeks and later. As in the retina, Syn expression in lens is also selectively affected in WWS. The colocalization of Synemin with vimentin, GFA, or NF supports the idea that Syn is a key cross-linking protein that connects different cytoskeletal structures. Moreover, stagnant Syn expression in WWS retina and lens reinforces the notion of a significant role of this protein in morphogenesis. PMID- 14552892 TI - Glutamatergic activation of the amygdala differentially mimics the effects of audiogenic seizure kindling in two substrains of genetically epilepsy-prone rats. AB - Comparisons of neuronal network mechanisms in closely related inherited seizure models are providing novel insights into epileptogenic pathophysiology. Genetically epilepsy-prone rats (GEPRs) exist in two substrains that inherit long term susceptibility to behaviorally distinct audiogenic seizures (AGS). GEPR-3s exhibit generalized clonic AGS, while GEPR-9s exhibit generalized tonic AGS. After AGS kindling the tonic AGS of GEPR-9s is followed by generalized posttonic clonus (PTC), while the generalized clonic AGS is followed by facial and forelimb (F&F) clonus in GEPR-3s. PTC and F&F clonus are very rare in GEPRs before AGS kindling. The neuronal network subserving AGS in GEPR-9s lies exclusively in brainstem sites, but amygdala (AMG) and other sites are recruited into the network after AGS kindling. The present study attempted to mimic the effects of AGS kindling by bilaterally microinjecting subconvulsive doses of N-methyl-D aspartate (NMDA) into the AMG of nonkindled GEPRs. NMDA (10 nmol/side) microinjected into AMG reversibly induced susceptibility to F&F clonus immediately following generalized clonic AGS in most nonkindled GEPR-3s. NMDA (7.5 nmol/side), microinjected into AMG temporarily induced susceptibility to generalized PTC immediately following tonic AGS in most nonkindled GEPR-9s. No seizures were induced in normal rats by these treatments, and no seizures were seen in GEPRs with these NMDA doses except those induced by acoustic stimuli. These findings support a critical role in AGS kindling for the AMG in the neuronal networks for both forms of AGS. However, the behavioral effect of the treatment was different in the two AGS substrains, suggesting interrelated but not identical pathophysiological mechanisms in these closely related epilepsy models. PMID- 14552891 TI - Gene transfer of glial cell line-derived neurotrophic factor promotes functional recovery following spinal cord contusion. AB - Neuronal cell death and the failure of axonal regeneration cause a permanent functional deficit following spinal cord injury (SCI). Administration of recombinant glial cell line-derived neurotrophic factor (GDNF) has previously been reported to rescue neurons following severe SCI, resulting in improved hindlimb locomotion in rats. In this study, thus, GDNF gene therapy using an adenoviral vector (rAd-GDNF) was examined in rats following SCI induced by dropping the NYU weight-drop impactor from a height of 25 mm onto spinal segment T9-T10. To evaluate the efficacy of intraspinal injection of recombinant adenovirus into the injured spinal cord, we observed green fluorescent protein (GFP) gene transfer in the contused spinal cord. GFP was effectively expressed in the injured spinal cord, and the most prominently transduced cells were astrocytes. The expression of GDNF was detected only in rats receiving rAd-GDNF, not the controls, and remained detectable around the injured site for at least 8 days. Open-field locomotion analysis revealed that rats receiving rAd-GDNF exhibited improved locomotor function and hindlimb weight support compared to the control groups. Immunohistochemical examination for the neuronal marker, calcitonin gene-related peptide (CGRP), showed an increase in CGRP+ neuronal fibers in the injured spinal cord in rats receiving rAd-GDNF treatment. Collectively, the results suggest that adenoviral gene transfer of GDNF can preserve neuronal fibers and promote hindlimb locomotor recovery from spinal cord contusion. This research should provide information for developing a clinical strategy for GDNF gene therapy. PMID- 14552893 TI - c-fos and tyrosine hydroxylase expression after an excitotoxic lesion on the nigrostriatal system: a study on the effects of hypoxia used as a preconditioning stimulus. AB - Hypoxia-ischemia during the perinatal period causes excitotoxic lesions in sensitive brain areas, such as the striatum. The impact of hypoxia-ischemia on nigral neurons is less well known. Hypoxia alone, a less traumatic event without overt histological sequelae, has neuroprotective properties when used as a preconditioning stimulus. In some pathologies, injured neurons of the nigrostriatal system in the adult may be the result of neurodegenerative processes that originated at early stages of life. The effects of hypoxia on the immunoreactivity to tyrosine hydroxylase of the dopaminergic neurons of the substantia nigra pars compacta and the effects of a period of hypoxia previous to an excitotoxic lesion were examined by means of histological and Western blot methods, at immediate and late periods of the episode. By counting the number of tyrosine hydroxylase-stained neurons and c-fos-positive nuclei a short period after injection of quinolinic acid into the striatum, we observed that hypoxia induced a more marked decrease in the number of tyrosine hydroxylase-stained neurons. On the contrary, c-fos-positive profiles decreased in the substantia nigra pars reticulata of the quinolinic acid-injected animals after the preconditioning hypoxia. Hypoxia alone did not affect the number of tyrosine hydroxylase-positive neurons in the pars compacta nor did hypoxia induce c-fos expression in the pars reticulata. More sensitive Western blot analysis of tissue blocks that included the whole substantia nigra demonstrated the same trend as the immunohistochemical results. We conclude that the responses of the substantia nigra neurons to hypoxia are regionalized and potential neuroprotective effects may depend on the vulnerability of each neuronal type. PMID- 14552894 TI - Epileptogenic activity of granulomas associated with murine cysticercosis. AB - Neurocysticercosis, caused by Taenia solium, is a common cause of neurologic disease in developing countries and among immigrants to the United States. Seizures are the most common clinical manifestation of neurocysticercosis. Imaging studies of patients with seizures from neurocysticercosis typically reveal evidence of an inflammatory reaction associated with the parasite or calcified granulomas. This study investigated whether a substance produced by the host granulomatous reaction to the dying parasite, in a mouse model of the infection, is sufficient to induce epileptiform activity. Granulomas associated with Taenia crassiceps cysticerci were removed from the peritoneal cavity of infected mice. One piece of the granuloma was used for blinded histological staging of the dying parasite. The second piece was used to generate extracts, which were injected into the hippocampus of an anesthetized Sprague-Dawley rat. Positive controls included animals injected with kainic acid, picrotoxin, or bicuculline. Seizures were recorded after injection of extracts from 6 out of 6 early stage granulomas, but only 1 out of 9 late stage granulomas. Injections of buffered saline, extracts from non-stimulated mouse spleen cells, and homogenates of viable parasite material caused no epileptiform activity. The data suggest that a substance in the granulomas early in the inflammatory response to the dying parasite is capable of inducing seizure activity. Further experiments are needed to dissect out the exact seizure mediator in the granuloma extracts. PMID- 14552895 TI - betaII-tubulin and GAP 43 mRNA expression in chronically injured neurons of the red nucleus after a second spinal cord injury. AB - Regeneration by chronically injured supraspinal neurons is enhanced by treatment of a spinal cord lesion site with a variety of neurotrophic and growth factors. The removal of scar tissue, with subsequent reinjury of the spinal cord, is necessary for injured axons to access tissue transplants placed into the lesion to support axon regrowth. The present study examined chronically injured and reinjured rubrospinal tract (RST) neurons to determine if changes in gene expression could explain the failure of these neurons to regenerate without exogenous trophic factor support. Adult female rats were subjected to a right full hemisection lesion via aspiration of the cervical level 3 spinal cord. Using radioactive cDNA probes and in situ hybridization, RST neurons in the contralateral red nucleus were examined for changes in mRNA levels of betaII tubulin and GAP 43 in an acute injury period (6 h-3 days), a chronic injury period (28 days after spinal cord injury (SCI)) and following a second lesion of the chronic injury site (6 h-7 days). Based upon the analysis of gene expression in single cells, GAP-43 mRNA levels were increased as early as 1 day following the initial SCI, but were no different than uninjured control levels at 28 days postoperative (dpo). The response to relesion was more rapid and higher than that detected after the initial injury with a significant increase in GAP 43 mRNA at 6 h that was maintained for at least 7 days. betaII-tubulin mRNA levels remained unchanged until 3 days after an acute injury followed by a decrease in expression to 30% below uninjured control values at 28 dpo. The expression of betaII-tubulin mRNA was significantly higher within 6 h after a second injury, where it remained stable for 5 days before a second increase occurred at 7 days after reinjury of the spinal cord. Thus, neurons in a chronic injury state retain the ability to respond to a traumatic injury and, in fact, neurons subjected to a second injury exhibit a significantly heightened expression of regeneration-associated genes. PMID- 14552896 TI - A herpesvirus vector can transduce axotomized brain neurons. AB - If gene therapy is to be used to promote axon regeneration after spinal cord injury, a suitable vector for transgene delivery must be obtained. Replication defective herpes simplex virus (HSV) vectors are promising candidates. We have examined whether they can express a LacZ transgene in injured neurons of adult rat brain. We transected the medial forebrain bundle, injected replication defective HSV/LacZ vectors close to the lesion site, and looked for transgene expression at 2-14 days after the lesion. The vectors carried the LacZ transgene controlled either by the cytomegalovirus immediate-early promoter (vector CS5) or the HSV latency-associated promoter (vector CS1). CS5 transfected many cells near the lesion at 2 days, but did not give persistent expression at 5 days. CS1, in contrast, labeled many neurons in midbrain regions remote from the injection site at 5 days, and much of this expression remained at 12-14 days. The neurons of most interest were in the substantia nigra pars compacta and parabrachial nuclei, which were axotomized by the lesion. Vector-driven beta-galactosidase expression was detected in neurons in both regions. These were confirmed as axotomized by double immunofluorescence for c-Jun. By 12-14 days, many substantia nigra neurons had disappeared but some transduced neurons remained; there was no net loss of transduced neurons from the parabrachial nuclei. These results show that an HSV vector is capable of transducing axotomized cells in the central nervous system and producing transgene expression in them for at least 2 weeks after injection. PMID- 14552897 TI - Age-related neurodegenerative changes in the central nervous system of estrogen deficient follitropin receptor knockout mice. AB - Age-related neurodegenerative conditions are characterized by neuronal death and degeneration that lead to a progressive functional decline. Among the factors influencing degenerative processes during aging are altered levels of neurotrophic ovarian steroid 17beta-estradiol (E2). The follitropin receptor knockout (FORKO) female mouse displays hormonal imbalance characterized by very low levels of circulating E2 and high levels of testosterone. FORKO mice (24 days and 20 months) were used to investigate structural and functional changes in the central nervous system. We now show that the lifelong depletion of the sex hormone E2 in female FORKO mice correlates with abnormal behavior associated with defined alterations in brain morphology early in life, especially in aged animals. Immunohistochemical studies showed significant increases in the size and number of immunoreactive glial fibrillary acidic protein glial cells found in several brain regions (cortex and hippocampus) and a dramatic decline in estrogen receptors alpha and beta in the amygdala of FORKO females. These changes were associated with increased signs of anxiety in these animals. In the present study, we provide evidence that the chronic depletion of sex hormone E2 from early development leads to neural impairments in adult and aged FORKO mice that are associated with hypertrophy of glial cells, cell loss in distinct brain regions, and abnormal behavior. We suggest that the hormonal imbalance found in the female FORKO mouse provides an experimental paradigm for the study of morphological correlates of the behavioral changes that often accompany menopause in women. PMID- 14552898 TI - Neuroprotection against NMDA excitotoxicity by group I metabotropic glutamate receptors is associated with reduction of NMDA stimulated currents. AB - The neurotransmitter glutamate can have both excitotoxic and protective effects on neurons. The excitotoxic effects have been intensively studied, whereas the protective effects, including the involvement of metabotropic glutamate receptors (mGluRs), remain unclear. In the present study, we tested the protective effects of the group-I-mGluR agonist (S)-3,5-dihydroxyphenylglycine (DHPG) on organotypic hippocampal slice cultures exposed to excitotoxic concentrations of N-methyl-D aspartate (NMDA). Effects of DHPG on electrophysiological responses induced by NMDA receptor activation were also recorded. Experiments were performed on organotypic hippocampal slice cultures derived from 7-day-old rats, with cellular uptake of propidium iodide as a marker for neuronal cell death. Slice cultures pretreated with DHPG (10 or 100 microM) for 2 h prior to exposure to 50 microM NMDA for 30 min displayed reduced propidium iodide uptake, compared to cultures exposed to NMDA only. The neuroprotective effect was confirmed by Hoechst 33342 staining, where the appearance of pycnotic nuclei after NMDA treatment was prevented by the DHPG pretreatment. Using caspase-3 activity to monitor the presence of apoptosis, failed to demonstrate this type of cell death in CA1 after NMDA application. The protective effect of DHPG was abolished by the mGluR1 selective antagonist (S)-(+)-alpha-amino-4-carboxy-2-methylbenzeneacetic acid (LY367385; 5 or 10 microM), whereas the mGluR5-selective antagonist 2-methyl-6 phenylethynylpyridine (MPEP; 1 microM) had no effect. Voltage-clamping of CA1 pyramidal cells in cultures treated with 10 microM DHPG for 2 h showed a significant depression of NMDA-induced inward currents compared to untreated controls. We conclude that neuroprotection induced by activation of group-I mGluRs involve mGluR1 and is associated with decreased NMDA-stimulated currents. PMID- 14552899 TI - Four PGE2 EP receptors are up-regulated in injured nerve following partial sciatic nerve ligation. AB - We previously reported that cyclooxygenase 2 (COX2) is up-regulated in macrophages in injured nerve of rats with partial sciatic nerve ligation (PSNL) and that local injection of the COX inhibitor ketorolac reversed tactile allodynia (Eur. J. Neurosci. 15: 1037-1047, 2002). These findings suggest that prostaglandins (PGs) are overproduced in injured nerve and are involved in the pathogenesis of neuropathic pain. In this study, we examined whether overproduced PGs alter the expression of PGE2 receptors, EP1-EP4, in injured nerve of PSNL rats. We found that cell profiles immunoreactive (IR) for four EP receptors, EP1, EP2, EP3, and EP4, are dramatically increased in injured nerve 2 and 4 weeks after PSNL. EP4-IR cells were the most abundant among these receptor-expressing cells. Immunoreactivities of all four EP receptors were localized to the cell nucleus. These EP-IR cells were never found in uninjured nerve. More than 80% EP1 and about 30% EP4-IR cells were identified as infiltrating macrophages since they coexpressed ED1. Only 3% EP2- and 6% EP3-IR cells coexpressed ED1. These findings suggest that majority of EP2-, EP3-, and EP4-IR cells are other types of inflammatory cells than macrophages. About 48% of macrophages expressed EP1 and 45% expressed EP4. Only 3 and 6% of macrophages, respectively, expressed EP2 and EP3. Perineural injection of ketorolac reversed tactile allodynia and suppressed the up-regulation of EP1 and EP4, but not the recruitment of ED1-IR marcrophages, in injured nerve. Our data suggest that following PSNL, PGE2 is one of the possible PGs overproduced in injured nerve and PG overproduction is involved in the up-regulation of EP receptors in injured nerve. PMID- 14552900 TI - Transient up-regulation of retinal EphA3 and EphA5, but not ephrin-A2, coincides with re-establishment of a topographic map during optic nerve regeneration in goldfish. AB - Eph tyrosine kinase receptors and their ligands, the ephrins, play a key role in the establishment of retinotectal topography during development. Tectal up regulation of ephrin-A2 in goldfish, coincident with the reestablishment of a retinotectal map, suggests a similar role during optic nerve regeneration. Here we report a complementary study of EphA3, EphA5 and ephrin-A2 expression in the retina. EphA3 and EphA5 are transiently up-regulated as ascending naso-temporal gradients, whereas ephrin-A2 remains uniform. The expression profiles differ from those in developing chick and mouse, suggesting that different combinations of retinal Eph receptors and ligands can generate topographic guidance information. PMID- 14552901 TI - Modification of hippocampal neurogenesis and neuroplasticity by social environments. AB - Synaptic plasticity and neurogenesis in the brain are affected by environmental stimuli. The present study was designed to investigate the effects of social environments on learning and memory, neurogenesis, and neuroplasticity. Twenty two-day-old rats were housed in isolation or in groups for 4 or 8 weeks and injected intraperitoneally with bromodeoxyuridine to detect proliferation among progenitor cells. The animals were also tested for learning in a water maze and for hippocampal CA1 long-term potentiation in vivo and in vitro. The results show that the number of newborn neurons in the dentate gyrus and the learning in a water maze decreased significantly in rats reared in isolation for 4 or 8 weeks, as compared with grouped controls. Induction of long-term potentiation in the CA1 area of rat hippocampus in vivo and in vitro was also significantly reduced by isolation. Furthermore, the effects of isolation rearing on spatial learning, hippocampal neurogenesis, and long-term potentiation could be reversed by subsequent group rearing. These findings demonstrated that social environments can modify neurogenesis and synaptic plasticity in adult hippocampal regions, which is associated with alterations in spatial learning and memory. PMID- 14552902 TI - Glial cell line-derived neurotrophic factor and brain-derived neurotrophic factor sustain the axonal regeneration of chronically axotomized motoneurons in vivo. AB - In contrast to injuries in the central nervous system, injured peripheral neurons will regenerate their axons. However, axotomized motoneurons progressively lose their ability to regenerate their axons, following peripheral nerve injury often resulting in very poor recovery of motor function. A decline in neurotrophic support may be partially responsible for this effect. The initial upregulation of glial cell line-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF) by Schwann cells of the distal nerve stump after nerve injury has led to the speculation that they are important for motor axonal regeneration. However, few experiments directly measure the effects of exogenous BDNF or GDNF on motor axonal regeneration. This study provided the first direct and quantitative evidence that long-term continuous treatment with exogenous GDNF significantly increased the number of motoneurons which regenerate their axons, completely reversing the negative effects of chronic axotomy. The beneficial effect of GDNF was not dose-dependent. A combination of exogenous GDNF and BDNF on motor axonal regeneration was significantly greater than either factor alone, and this effect was most pronounced following long-term continuous treatment. The ability of GDNF, either alone or in combination with BDNF, to increase the number of motoneurons that regenerated their axons correlated well with an increase in axon sprouting within the distal nerve stump. Thus long-term continuous treatment with neurotrophic factors, such as GDNF and BDNF, can be used as a viable treatment to sustain motor axon regeneration. PMID- 14552903 TI - Multi-joint movement of the cat hindlimb evoked by microstimulation of the lumbosacral spinal cord. AB - Microstimulation of the lumbosacral spinal cord may be an effective tool for the restoration of locomotion after spinal cord injury. To examine this possibility, complex coordinated multi-joint hindlimb movements were evoked by electrical stimulation with sine waveform modulation using a single microelectrode positioned in the L5-S1 spinal cord. Four types of hindlimb movement (flexion, extension, abduction, and adduction) were identified, and their stimulation locations were mapped onto cross-sectional drawings of L5-S1 spinal cord following histological examination of electrode tracks in the cord. Hindlimb flexion was evoked without abduction/adduction at many locations in the dorsal part of the L5-S1 spinal cord, whereas extension was evoked with abduction/adduction in the ventral part of the cord. Bilateral reciprocal lifting of the hindlimb was evoked by implanting two microelectrodes (one on each side) in the spinal cord. This study indicates that functional hindlimb movements can be elicited by activating a small number of sites in lumbosacral spinal cord. PMID- 14552904 TI - Sex-specific KCC2 expression and GABA(A) receptor function in rat substantia nigra. AB - GABA(A) receptor activation by muscimol has sex and age specific effects on substantia nigra reticulata (SNR)-mediated control of generalized seizures. GABA(A) receptor agonists depolarize or hyperpolarize neurons depending upon the level of expression of the neuronal specific potassium chloride contransporter KCC2. We studied KCC2 mRNA expression in the SNR as a function of sex and age and correlated KCC2 expression with the in vivo and in vitro effects of muscimol. Methods included in situ hybridization, gramicidin-perforated patch clamp and fura-2 AM imaging of acute SNR slices. KCC2 mRNA expression increased between postnatal days (PN) 15 and 30 in both sexes, and reached adult levels in males by PN30. Female PN15 and PN30 SNR neurons contained more KCC2 mRNA compared with age matched males. In male PN14-17 rats, bath application of the GABA(A) receptor agonist muscimol in acute SNR slices depolarized neurons and increased intracellular calcium concentration ([Ca(2+)](i)). Furthermore, acute in vivo administration of muscimol upregulated, whereas blockade of L-type voltage sensitive calcium channels with nifedipine downregulated KCC2 mRNA. In contrast, in female PN14-17 rats, bath application of muscimol hyperpolarized SNR neurons and did not alter [Ca(2+)](i). In vivo muscimol administration acutely downregulated KCC2 mRNA expression whereas nifedipine had no effect. The lower expression of KCC2 mRNA in infantile male SNR neurons may explain why muscimol induced depolarization and [Ca(2+)](i) increases occur only in males. Consequently, GABA(A) receptor activation selectively upregulates the expression of calcium-regulated genes, such as KCC2, in male SNR, promoting the sexual differentiation of the SNR. PMID- 14552905 TI - Regionally specific modulation of brain apolipoprotein E in the mouse during the estrous cycle and by exogenous 17beta estradiol. AB - Studies have suggested that 17beta estradiol (E2) can modify apolipoprotein E (apoE) expression. The current study determined if apoE protein varied in different regions of the mouse brain as a function of the estrous cycle and if E2 could increase apoE protein expression. In this study apoE concentration was lowest on estrus in the hippocampus, cingulate cortex and frontal cortex. In contrast, apoE concentration was highest on estrus in the olfactory bulb and cerebellum. There were no differences in the striatal apoE expression throughout the estrous cycle. Exogenous E2 significantly raised tissue levels of apoE in the olfactory bulb and cerebellum at 5 days after treatment. There was a slight, but nonsignificant increase in cortical expression of apoE and no change in striatum. Immunocytochemical localization studies found estrogen receptor alpha (ERalpha) and estrogen receptor beta (ERbeta) in cortical neurons and glia. In the cerebellum and olfactory bulb, ERbeta was seen primarily in glia. ERalpha was not observed in the cerebellum and was rare in the olfactory bulb. Neither ERalpha nor ERbeta was seen in the striatum. Our data show regional differences in the production of apoE throughout the estrous cycle. In addition, exogenous E2 has regionally specific effects on apoE expression. Regional variability in apoE production appears to vary as a function of the estrogen receptor subtype. PMID- 14552907 TI - Survival and differentiation of adult rat-derived neural progenitor cells transplanted to the striatum of hemiparkinsonian rats. AB - We investigated the survival, distribution and differentiation capabilities of adult rat hippocampus-derived progenitor cells (AHPs) by grafting them into either the intact or dopamine (DA)-denervated adult rat striatum (ST). Furthermore, we tested the effects of the in vivo administration of retinoic acid (RA) on the differentiation of the grafted cells. AHPs, prelabeled in vitro with bromodeoxyuridine (BrdU) and primed with RA, were transplanted bilaterally into the ST of hemiparkinsonian rats. Twenty animals were divided in four groups: three groups received i.p. injections of RA (1.5 mg/kg/day) for 1, 2 or 4 weeks and one group received vehicle injections for 4 weeks. Approximately 60% of the implanted BrdU-immunoreactive (BrdU+) cells were present in either intact or lesioned ST after 5 weeks of transplantation, with a striking widespread radial distribution from the implantation site. The cells became morphologically integrated with the surrounding host tissue, with no evidence of tumor formation. Approximately 18% of the BrdU+ cells were immunoreactive for the glial precursor marker NG2 and occasionally BrdU+ cells co-expressed the neuronal marker TuJ1. This differentiation pattern was similar in the intact and DA-denervated ST. Although further research is needed to find more adequate methods to drive the differentiation of these cells toward the desired phenotypes, the survival, differentiation potential and widespread distribution throughout the ST observed in this study suggest that AHPs may be useful in treatment of degenerative disorders affecting the nervous system. PMID- 14552906 TI - Overexpression of spermidine/spermine N1-acetyltransferase elevates the threshold to pentylenetetrazol-induced seizure activity in transgenic mice. AB - Activation of polyamine catabolism in transgenic mice through an overexpression of spermidine/spermine N(1)-acetyltransferase (SSAT) results in a massive overaccumulation of the diamine putrescine in most tissues including brain. Putrescine pool in transgenic animals was strikingly expanded in every six brain regions analyzed at present. Pons (23-fold), cerebellum (37-fold), cerebrum (34 fold), and hippocampus (16-fold) showed the greatest increases in putrescine levels. Moreover, the molar ratio of putrescine to spermidine was increased in the different brain regions of the transgenic animals on an average of nearly 40 fold. Upon an exposure of the animals to pentylenetetrazol (PTZ) infusions, a compound known to induce epilepsy-like seizure activity, the SSAT transgenic mice showed significantly elevated seizure threshold to both clonic and tonic convulsions in comparison with their syngenic littermates. This difference, however, disappeared when the animals were treated with ifenprodil prior to PTZ infusions. The latter compound acts as an antagonist of N-methyl-D-aspartate receptor by binding to the polyamine site of the receptor. Overexpression of SSAT likewise appeared to protect the transgenic animals from PTZ-induced neuron loss in the hippocampus. As putrescine is known to serve as a precursor to gamma aminobutyric acid (GABA), we carried out (1)H NMR analyses the results of which revealed that the levels of the inhibitory amino acid GABA and its excitatory counterpart glutamate were indistinguishable in syngenic and transgenic animals in all brain regions analyzed. The present results suggest that the frequently observed enhanced accumulation of putrescine in response to brain insults belongs to neuroprotective measures rather than being a cause of the subsequent injury. PMID- 14552908 TI - GM1 enhances dopaminergic markers in the brain of aged rats. AB - A number of presynaptic markers are compromised in the dopaminergic neurons of aged Sprague-Dawley rats (22 months old) compared with young rats (3 months old). Indeed, in the striatum of the aged rats there is a diminished capacity to transport dopamine (DA), to bind the dopamine transporter (DAT) marker mazindol, to bind the vesicular monoamine transporter 2 (VMAT2) marker dihydrotetrabenazine, and to release DA under basal conditions or after induction by K(+) or amphetamine. Furthermore, the expression of DAT and VMAT2 mRNA in the midbrain is suppressed. GM1 ganglioside, 30 mg/kg ip daily, administered for 30 days, restores the afore-mentioned markers to values approaching those for young rats. Taken together with our published observations that GM1 partially restores tyrosine hydroxylase activity and DA metabolism in aged nigrostriatal and mesoaccumbal neurons and improves their morphology, our work suggests that GM1 might act as a dopaminergic neurotrophic factor in the aged brain and be a useful adjuvant for treating age-associated dopaminergic deficits. PMID- 14552909 TI - Behavioral and neurochemical characterization of transgenic mice carrying the human presenilin-1 gene with or without the leucine-to-proline mutation at codon 235. AB - Human presenilin-1 (PS1) mutations are associated with the incidence of familial Alzheimer's disease. The present study evaluated the behavioral and neurochemical effects of the L235P mutation (substitution of leucine by proline at codon 235) of the human PS1 gene, which has been linked to a form of early-onset Alzheimer's disease. Except for a significant increase in the production of beta-amyloid-42, the mutant mice did not show any overt signs of Alzheimer-like neuropathology in the form of plaque formation, changes in choline acetyltransferase activity, or somatostatin content in the brain. Cognitive assays indicated that the mutation did not affect the acquisition or reversal of a spatial reference memory task in the water maze or performance on a spatial working memory task. In contrast, L235P PS1 transgenic mice exhibited a significant impairment in a test of spontaneous object recognition. This dissociation is suggestive of a preferential impairment of the extrahippocampal memory system and is consistent with what has been reported in another pathological mutation (substitution of leucine by valine at codon 286) of the PS1 gene. PMID- 14552910 TI - Glucose deprivation produces a prolonged increase in sensitivity to glutamate in cultured rat cortical neurons. AB - In this study we investigated whether the link between mitochondrial dysfunction and deregulation of Ca(2+) homeostasis preceding excitotoxic cell death is mediated by cellular deenergization. Glycolytic and/or mitochondrial ATP synthesis was inhibited with 2-deoxy-D-glucose (2DG) and oligomycin, respectively. Changes in cytoplasmic Ca(2+) concentration ([Ca(2+)](c)) and mitochondrial membrane potential were simultaneously measured in response to low (10 microM) glutamate concentrations, using the fluorescence dyes fura-2FF and rhodamine 123. 2DG, which blocks glycolysis and also inhibits mitochondrial respiration due to depletion of pyruvate, greatly increased and accelerated glutamate-induced elevation of [Ca(2+)](c) and mitochondrial depolarization. The 2DG-induced hypersensitivity to glutamate was observed even after 150-min washout of 2DG with glucose-containing medium, suggesting a permanent deterioration of mitochondrial function. Prior blockade of only glycolytic (2DG with pyruvate) or only mitochondrial (oligomycin) ATP synthesis did not affect neuronal sensitivity to glutamate. Collectively, these studies show that to maintain the sensitivity of neurons to glutamate at control levels at least one of the cellular sources of ATP production must be intact. Either glycolysis or oxidative phosphorylation can effectively support Ca(2+) homeostasis in cultured forebrain neurons. PMID- 14552911 TI - Toe out angle: a functional index for the evaluation of sciatic nerve recovery in the rat model. AB - In experimental peripheral nerve studies, the rat sciatic nerve model is widely used to examine functional outcome following nerve injury and repair. A variety of evaluation methods exist in the literature, but an adequate selection continues to be a critical point for the researcher. Rats with sciatic nerve injury typically ambulate with an external rotation of the foot. A new functional assessment instrument, the toe out angle (TOA) is quantified using computerized gait analysis. We compared Sciatic Functional Index (SFI) with TOA parameter after peripheral nerve transection and entubulation repair. We found a good correlation between SFI and TOA measurements in terms of predicting functional recovery. Moreover, the TOA provides information on the biomechanical consequences of the external rotation of the foot in the stance phase of walking. PMID- 14552912 TI - Neuroprotective effects of pyruvate in the quinolinic acid rat model of Huntington's disease. AB - The neuroprotective effects of pyruvate, the end metabolite of glycolysis, were studied in an animal model of Huntington's disease (HD). Intrastriatal injection of quinolinic acid (QA) caused widespread damage to rat striatum as determined from cresyl violet staining and immunohistochemical analysis. Intraperitoneal administration of pyruvate at doses of 500-1000 mg/kg significantly reduced striatal lesions induced by QA. A lower pyruvate concentration of 250 mg/kg was not protective; however, quadruple applications at this dosage was effective in reducing lesion volumes. The protective effects of pyruvate were found over a range of times, from application at the time of QA injection to 1 h post administration; however, no protection was conferred if pyruvate was applied 30 min prior to QA injection or 3 h post-administration. We also found pyruvate protects different types of striatal neurons against QA toxicity including GABAergic projection neurons, cholinergic interneurons and NADPH-diaphorase interneurons. These results suggest that pyruvate may be effective in reducing neuronal damage in HD. PMID- 14552932 TI - The prevention challenge: an overview of this supplement. PMID- 14552933 TI - New thinking on older adults' physical activity. PMID- 14552934 TI - Assessing the building blocks of function: utilizing measures of functional limitation. AB - To better understand the health of older people, it is valuable to go beyond conventional measures of disease and self-perceived health and utilize assessments of functional status and disability. Physical decrements can be characterized at the level of organs and body systems (impairments); the organism as a whole (functional limitations); and the person in the context of the environment and its challenges (disability). To understand the dynamics of the pathway leading from disease to disability, it is critical to measure functional limitations that can be assessed through either self-report or standardized objective measures of physical performance. Both of these approaches may be used to evaluate functions such as grasping objects, walking, and climbing stairs. In aging research, measures of functional limitation are utilized as outcomes that indicate the impact of disease, impairments, and other risk factors on function. In turn, measures of functional limitation can be used to characterize the functional status of individuals and populations, and are powerful predictors of various adverse outcomes, including incident disability in people not currently disabled. Functional limitation measures add substantially to our knowledge about older populations, but further work is needed to promote their standardization and use in both clinical and research settings. PMID- 14552935 TI - Assessing disability in studies on physical activity. AB - Despite the increased focus on disability as an important research outcome, there remains a need to improve disability instrumentation. This article discusses the need for improved operational differentiation in instruments used to assess disability outcomes. It details the challenge of clearly distinguishing between the concepts of function and disability in outcome instruments used in physical activity studies. The second issue in existing instruments is the limited precision to detect important changes in disability in response to physical activity interventions. Computer adaptive testing is discussed as a promising avenue for resolving the methodologic limitations inherent in existing outcome instruments. Priorities for future research that move the field forward in both of these areas are discussed. PMID- 14552936 TI - Does late-life physical activity or exercise prevent or minimize disablement? A critical review of the scientific evidence. AB - Physical activity and exercise are widely purported to enhance health and minimize or prevent functional loss and disability. Yet, do the benefits of late life physical activity or exercise extend beyond disease and impairment-level factors? Does late-life physical activity minimize or prevent functional limitations and disability? To address these questions, a best-evidence framework was used to examine the effects of late-life physical activity on disablement outcomes. This review shows that exercise-particularly walking-increases muscle strength and aerobic capacity and reduces functional limitations. It is less clear, however, whether physical activity or exercise prevents or minimizes physical disability. Furthermore, this review shows a discrepancy between prospective and experimental studies: several well-conducted prospective studies show a beneficial effect of physical activity on minimizing disability, whereas the majority of experimental studies that have examined disability as an outcome do not show improvements in disability. Three research priorities are identified that would advance the science in this field: (1). development of a clear conceptual and theoretical framework of late-life physical activity and assessment; (2). use of a disablement outcomes framework to examine the outcomes of late-life physical activity; and (3). development of a mechanism of action explaining the relationship between physical activity and exercise and disablement outcomes. PMID- 14552937 TI - Conceptual challenges in linking physical activity and disability research. PMID- 14552938 TI - The benefits of strength training for older adults. AB - Aging is associated with a number of physiologic and functional declines that can contribute to increased disability, frailty, and falls. Contributing factors are the loss of muscle mass and strength as age increases, a phenomenon called sarcopenia. Sarcopenia can result or be exacerbated by certain chronic conditions, and can also increase the burden of chronic disease. Current research has demonstrated that strength-training exercises have the ability to combat weakness and frailty and their debilitating consequences. Done regularly (e.g., 2 to 3 days per week), these exercises build muscle strength and muscle mass and preserve bone density, independence, and vitality with age. In addition, strength training also has the ability to reduce the risk of osteoporosis and the signs and symptoms of numerous chronic diseases such as heart disease, arthritis, and type 2 diabetes, while also improving sleep and reducing depression. This paper reviews the current research on strength training and older adults, evaluating exercise protocols in a variety of populations. It is clear that a variety of strength-training prescriptions from highly controlled laboratory-based to minimally supervised home-based programs have the ability to elicit meaningful health benefits in older adults. The key challenges as this field of exercise science moves forward are to best identify the most appropriate strength-training recommendations for older adults and to greatly increase the access to safe and effective programs in a variety of settings. PMID- 14552939 TI - Balance training to maintain mobility and prevent disability. AB - Balance is important for the safe performance of many activities that allow older people to remain independent in their community. Housework, cooking, shopping, and travel generally require the ability to stand, reach, turn, and bend down and pick up objects from the floor. Multiple interacting factors are implicated in the deterioration of balance. Several strategies have been tested to improve balance and reduce falls. Home-based individualized training that attempted to improve identified deficits reduced falls and improved physical performance and stabilized or reduced disability. Home-based exercise programs that included low intensity strength and balance training have improved balance and reduced fall rates by about 40% compared to controls. Class-based exercise programs in senior centers or exercise centers have improved balance and physical performance, and some have reduced falls. Programs such as tai chi and social dance look promising and should be further investigated. PMID- 14552940 TI - Balance and mobility challenges in older adults: implications for preserving community mobility. AB - Mobility is a critical component of most activities of daily living. Therefore, maintenance of mobility independence is essential for health and well-being. Balance control is a primary requirement for successful mobility. In this article, we first discuss the challenges to mobility and balance that older adults face within the community. Next, we examine the implications for preserving mobility, in particular the efficacy of different types of exercise based interventions that have been used. Exercise and training done in relatively sterile environments, such as a laboratory, do not simulate the environmental challenges one faces in the community and are not particularly beneficial. In addition, the outcome measure(s) used to evaluate the success of the intervention programs need to reflect improvement in functional mobility within the community. We propose a mobility envelope measured from excursions into the community over a week as a likely candidate for outcome measure. PMID- 14552941 TI - Challenging aging stereotypes: strategies for creating a more active society. AB - BACKGROUND: The myths of aging provide a partial explanation for why older people have not been primary targets for health promotion and disease-prevention programs. Accelerated population aging signals an urgent need for increased attention to health promotion and disease-prevention interventions across the entire life course. OBJECTIVE: The purpose of this article is to review what is known about: (1). the prevalence and nature of ageist stereotypes; (2). the varied ways in which ageist stereotypes are harmful to health, functioning, and well-being; and (3). strategies for effective communication with older adults around lifestyle issues, with particular emphasis on physical activity. RESULTS: This article reviews literature on current myths and reality of aging in the context of designing health promotion programs for older adults. Strategies for combating ageist stereotypes are based on a multilevel view of determinants of health and aging. CONCLUSIONS: Ageist stereotypes are pervasive in U.S. society and harmful to older adults' psychological well-being, physical and cognitive functioning, and survival. Concrete strategies for communicating with older adults can increase the effectiveness of health promotion programs. Strategies for combating ageism and creating a healthier society must address all segments of society, and include educational and media campaigns, an expansion of current research foci, greater sensitivity from care providers, more opportunities for intergenerational linkages, the design of productive roles for older adults, a retrofit of the built environment, and intensified and collaborative action from both the public and private sectors. PMID- 14552942 TI - Promoting physical activity for older adults: the challenges for changing behavior. AB - This article addresses the formidable topic of promoting physical activity among older adults. The primary focus is on older adults who are at risk for or who have evidence of functional limitations or physical disabilities that often accompany chronic disease; where relevant, findings about healthy older adults are also discussed. All of these individuals experience barriers for physical activity that range from cultural expectations to complex symptoms that accompany the disablement process. Conducting physical activity programs in diverse settings and utilizing multiple channels of communication help to alleviate some of these barriers. A major emphasis of the article concerns behavior change strategies necessary in interventions for at-risk older adults with diverse needs and individual differences. Based on recent investigations, several challenges for research are proposed. It is suggested that further attention should be directed toward individualized tailoring of programs that recognize unique barriers in older adults such as intermittent illness and the burden of caregiving. Furthermore, research on physical activity interventions should examine whether they will benefit from collaborative, social problem-solving models of behavior change that link individual and group-mediated interventions. The final challenge proposed is research to examine viable ways of linking these interventions to population-based health promotion activity programs. PMID- 14552943 TI - Promoting physical activity among older adults: from ecology to the individual. AB - The promotion of physical activity in older populations holds the promise of promoting health and reducing the risk of disablement. Individual- and community based programs are often presented as being distinct and, in some cases, antithetical. The objective of this paper is to illustrate, through the use of an ecologic model, that individual- and community-based programs are complementary. The following topics are discussed: (1). the development of transdisciplinary theories; (2). the establishment of better measures to address the linkages across individual behavior, group behavior, the built environment, and the temporal sequence of biological, behavioral, and environmental factors; (3). the translation of research into practice and policies; and (4). the training of a new generation of scholars and practitioners to develop and execute research, practice, and policy agendas in this area. An ecologic model fosters a comprehensive agenda based on biological, behavioral, and environmental factors. The study of the interaction of the individual with the environment is challenging but potentially more important than the study of the role of the individual or the environment alone in the promotion of physical activity. PMID- 14552944 TI - Active living for assisted living: promoting partnerships within a systems framework. AB - In response to a growing need for assistance among our aging population, assisted living facilities have been designed to fill the widening chasm between community living and nursing care. Although sedentary behavior has been linked to functional limitations and disability, no comprehensive information exists about the social and physical environments and the programming available to promote physical activity in assisted living. Accordingly, this article includes data from an exploratory study that underscores the issues clearly related to physical activity for older adults in assisted living. The intent of this exploratory study was to partner with executive directors in order to conduct an analysis of the social and physical characteristics of assisted living. Interviews were conducted with executive directors (N=21) to discuss methods for assessing and promoting positive lifestyle behaviors with distinct emphasis on the targeted behavior of physical activity. Potential ways in which the social and physical environments could be modified to promote and support physically active living were identified. Clearly, promoting physical activity in assisted living is a challenge and will require a partnership with assisted living communities to develop effective and feasible systems-based interventions designed to make environments more engaging and, thereby, promote active living. PMID- 14552945 TI - Medical assessment for health advocacy and practical strategies for exercise initiation. AB - The universal caution to consult your family doctor before beginning an exercise program creates an image of exercise as potentially harmful. Moreover, insistence on extensive screening prior to exercise is both unrealistic and often inappropriate for the older adult. Recasting the role of the physician as a physical activity advocate rather than as a gatekeeper is recommended for incorporation into guidelines for exercise screening. A geriatric assessment focusing on identification of specific parameters predictive of disablement risk can be incorporated into the exercise prescription as a guide for initiating exercise. The purpose of this article is to: (1). review the role of the primary care physician in screening and advocating exercise; (2). examine objectively the risk of exercise among older adults and place these risks in context with current screening guidelines with a particular emphasis on the exercise test; and (3). examine how key concepts derived from epidemiologic studies of disability can be distilled into practical guidelines for exercise therapy. We provide an overview of relevant literature related to screening and initiating exercise. Key challenges are highlighted and discussed. Suggestions for changes in policy are recommended. Given the apparent discordance between screening guidelines for the older adult and risk of adverse events, and between existing recommendations for physical activity and epidemiologic studies of disability, an evidence-based approach is recommended to review and revise screening and prescribing practices. PMID- 14552946 TI - The National Blueprint Consensus Conference summary report: strategic priorities for increasing physical activity among adults aged >or=50. AB - This report summarizes the outcome of the National Blueprint Consensus Conference that was held in October 2002. At this conference, representatives of more than 50 national organizations convened in Washington DC with the goal of identifying high-priority and high-feasibility strategies that would advance the National Blueprint and that could be initiated within the next 12 to 24 months. The National Blueprint Consensus Conference has identified an ambitious agenda of 18 strategies that will need to be implemented in order to overcome societal barriers to physical activity among the middle-aged and older adult population. National organizations charged with the task of implementing the high-priority strategies will use professional networks, established delivery channels, and communication systems to translate the blueprint strategies into action. PMID- 14552947 TI - Physical activity to prevent or reverse disability in sedentary older adults. PMID- 14553797 TI - Liver fibrosis grade classification with B-mode ultrasound. AB - B-mode images of 20 fresh postsurgical human liver samples were obtained to evaluate ultrasound ability in determining the grade of liver fibrosis. Image features derived from gray level concurrence and nonseparable wavelet transform were extracted to classify fibrosis with a classifier known as the support vector machine. Each liver sample subsequently underwent histologic examination and liver fibrosis was graded from 0 to 5 (i.e., six grades total). The six grades were then combined into two, three, four and six classes. Classifications with the extracted image features by the support vector machine were tested and correlated with histology. The results revealed that the best classification accuracy of two, three, four and six classes were 91%, 85%, 81% and 72%, respectively. Thus, liver fibrosis can be noninvasively characterized with B-mode ultrasound, even though the performance declines as the number of classes increases. The elastic constants of 16 samples out of a total of 20 were also correlated with the image features. The Pearson correlation coefficients indicated that the image features are more strongly correlated with the fibrosis grade than with the elastic constant. PMID- 14553798 TI - Ureteric jet in the anaesthetised child. AB - We have already suggested that there is an active bimodal sphincteric mechanism in the human vesicoureteral junction (VUJ). This mechanism is affected by immaturity, hormonal effects and surgical ablation. This study set out to investigate the effects of anaesthesia on this mechanism. A total of 16 children undergoing surgery were recruited. They were well hydrated. The ureteric jets before and after anaesthesia were recorded using the colour and pulse wave Doppler. Of the total, 14 children showed mature (complex pattern) and two showed immature (monophasic pattern) waveforms before surgery. After anaesthesia, all showed the monophasic waveform (p=0.01). Thus, the sphincteric action of VUJ as manifest by the complexity of the ureteric jet was modified by anaesthesia by loss of the complex pattern. This supports the hypothesis that the complex pattern is a neural modulation of the simpler myogenic mechanism that produces the monophasic jet. PMID- 14553799 TI - Doppler ultrasonographic evaluation of hepatic blood flow in clinical sepsis. AB - The aim of this study was to determine the hemodynamic response of the liver to sepsis by measuring hepatic blood flow. Thirty patients with sepsis were examined using Doppler ultrasonography and measurements of portal venous blood flow, hepatic arterial blood flow and total hepatic blood flow were recorded. Data were retrospectively reviewed and compared with findings for a control group of 12 healthy subjects. Significantly increased values of hepatic blood flow (p<0.01) and portal venous blood flow (p<0.001) were observed in patients during early sepsis (5 h). In contrast, hepatic arterial blood flow was not significantly different from controls. During late sepsis (24 h), no significant differences were observed between patients and healthy controls. Our results support the concept that hepatic blood flow is significantly increased in patients with early sepsis due to increased portal venous blood flow; however, during late sepsis, hepatic blood flow is similar to that in controls. PMID- 14553800 TI - Perfusion harmonic imaging in acute middle cerebral artery infarction. AB - Initial reports indicate that cerebral perfusion deficits in acute ischemic stroke might be detectable by means of transcranial harmonic imaging after an ultrasound contrast agent (UCA) bolus injection. Twenty-four patients with acute middle cerebral artery (MCA) infarction were investigated twice with perfusion harmonic imaging (PHI) after Levovist (Schering, Berlin, Germany) bolus injection no longer than 12 h after symptom onset. The findings were compared with those of cranial computed tomography (CCT). All 24 patients suffered from acute ischemic stroke of the MCA territory (median National Institutes of Health Stroke Scale score: 15 points). Corresponding to the area of infarction in follow-up CCT, a marked contrast deficit was visualized in 19 of 24 patients by initial PHI, which had a sensitivity and specificity of 86.4% and 96.2%, respectively, for predicting the occurrence and localization of a definite infarction in the midthalamic plane. The area of hypoperfusion in the initial PHI investigation correlated with the definite area of infarction in follow-up CCT (r=0.66, p<0.01). When time-intensity curves of both hemispheres were compared, the areas under the curve were significantly less in the symptomatic brain regions (p=0.01). With PHI and UCA bolus injection, it is possible to assess cerebral perfusion deficits that correlate with the definite area of infarction in acute ischemic stroke patients. PMID- 14553801 TI - Assessment of local changes of cerebral perfusion and blood concentration by ultrasound harmonic B-mode contrast measurement in piglet. AB - This study tested the hypothesis that changes in the blood concentration, and possibly in the perfusion, of different areas in the brain can be assessed by the use of ultrasound contrast agent (CA) and (linear) echo densitometry. The experiments were performed with piglets (n=3) under general anesthesia and artificial ventilation. Ultrasound CA was administered through a femoral vein as a short bolus. First passage wash-in curve was measured from image gray level during continuous low level (mechanical index<0.2) ultrasound imaging. This curve was obtained from 1-cm2 areas of the cortex (surface), the brain stem (inner) and the left carotid artery (vessel). Cerebral hemoglobin concentration changes were measured with near-infrared spectroscopy (NIRS). This approach enabled a cross validation of these techniques. The measurements were repeated under conditions of normocapnia, mild hypercapnia and deep hypercapnia. Several physiologic signals, as well as the carotid blood flow, were measured simultaneously and related to gray level by linear regression analysis. The most significant results found were a high R2-statistic of the regression of the percentage change of the peak of the surface and inner wash-in curves with the arterial carbon dioxide pressure (R2=0.63 and R2=0.70, respectively), the blood pH (R2=0.79 and R2=0.81), the carotid flow (R2=0.75 and R2=0.72) and the partial arterial oxygen pressure (R2=0.47 and R2=0.55). Finally, a high correlation of peak gray level with total hemoglobin concentration change, independently measured by NIRS, was found (R2=0.69). In conclusion, these experiments show a reasonable intersubject variability of various relative measures derived from gray level ultrasound wash in curves. High sensitivity to physiologic changes related to hypercapnia was observed for the peak contrast of wash-in curves. For up-slope and area-under-the curve (first passage) this was lower but still highly significant. The gray-level ultrasound measures are highly correlated to changes in regional hemoglobin concentration in brain tissue assessed by NIRS. PMID- 14553802 TI - Comparison of two-dimensional and three-dimensional ultrasound in the assessment of the cervix to predict preterm delivery. AB - This study sought to determine whether 3-D transvaginal ultrasound (3D-TVS), compared with the 2-D transvaginal approach (2D-TVS), offers a better identification of some specific features of the uterine cervix that could be useful in the prediction of spontaneous preterm delivery (PTD). A total of 103 women with singleton pregnancies were prospectively evaluated with 2D-TVS and 3D TVS in the second or third trimester of pregnancy. Statistical analysis was performed with Wilcoxon matched-pairs test, chi2 and Pearson test (p<0.05 was considered significant) and receiver operating characteristic (ROC) curve analysis. Significant differences between these approaches were found for cervical length (p<0.001). A significant correlation was identified between 3D TVS cervical length and the interval between ultrasound examination and delivery as well as the gestational age at delivery (both p<0.001). ROC curve identified a threshold for 3D-TVS cervical length of 35 mm as an optimal predictor of PTD in the second trimester (sensitivity 100%, specificity 88%). 3D-TVS evaluation of the cervix in pregnancy seems to be an effective, noninvasive method for predicting PTD risk. PMID- 14553803 TI - The assessment of normal fetal brain volume by 3-D ultrasound. AB - The assessment of the fetal brain volume (BV) is very important in the evaluation of fetal growth. The purpose of this study is to use the three-dimensional (3-D) ultrasound (US) in constructing reference centiles of the fetal BV during normal gestation for clinical application. This study was undertaken by a prospective and cross-sectional design. In total, 203 singleton fetuses ranged between 20 and 40 weeks of gestation and fit the criteria of normal pregnancies were enrolled in this study. Our results showed that fetal BV is highly correlated with the gestational age (GA). Furthermore, using GA as the independent variable and the fetal BV as the dependent variable, the best-fit regression equation was BV (mL)= 171.48036+4.8079xGA+0.29521xGA2 (r=0.99, n=203, p<0.0001), with SD of BV (mL)=1.2533x(9.9474+0.07133xGA). In addition, the common growth parameters of the fetal biometry, such as biparietal diameter (BPD), occipitofrontal diameter (OFD), head circumference (HC), abdominal circumference (AC), femur length (FL) and estimated fetal weight (EFW), were all highly correlated with the fetal BV (all p<0.0001). In conclusion, our data on the fetal BV assessed by 3-D US may serve as a useful reference in evaluating fetal growth. PMID- 14553804 TI - Three-dimensional power Doppler ultrasound for the assessment of the fetal brain blood flow in normal gestation. AB - Early identification of the abnormal fetal brain vascularization and blood flow is very important, because the deficient perfusion of the fetal brain may be related to a poor prognosis of the central nervous system (CNS) development. To assess the fetal brain vascularization and the blood flow in normal gestation, we measured the fetal brain vascularization and the brain blood flow in normal fetuses using three-dimensional (3-D) power Doppler ultrasound (US) and the quantitative 3-D power Doppler histogram analysis. This study was undertaken by a prospective and cross-sectional design. In total, 155 normal singletons with gestational age (GA) between 21 and 40 weeks were included. The 3-D power Doppler US and the quantitative 3-D histogram analyses were used to assess the fetal brain vascular indexes, i.e., vascularization index (VI), flow index (FI) and vascularization-flow index (VFI), in each case. Our results revealed that all the fetal brain VI, FI and VFI increased significantly with GA (all p<0.001). In addition, the fetal brain VI, FI and VFI were all significantly correlated with the common fetal growth indices, such as biparietal diameter, occipitofrontal diameter, head circumference, abdominal circumference, femur length and estimated fetal weight. Our study indicates that fetal brain vascularization and blood flow increase significantly with the advancement of GA as well as the fetal common growth indices during normal gestation. We believe our data may serve as a reference for further studies of the fetal brain blood flow in abnormal conditions. PMID- 14553805 TI - Doppler evaluation of peripheral vascular adaptations to transverse aortic banding in mice. AB - Transverse aortic banding in mice is commonly used to produce pressure overload, but the resulting cardiac hypertrophy is variable and the actual load produced is unknown. The purposes of the study were to characterize peripheral blood flow in banded mice using noninvasive Doppler methods, investigate whether changes in flow could predict the amount of cardiac hypertrophy induced and validate the simplified Bernoulli equation for estimating the pressure drop across the stenosis in very small vessels. Wild-type mice underwent aortic banding (n=15) or sham operation (n=6). Doppler velocity was measured in the right and left carotid arteries (RCA and LCA) 1 day later, and the heart weight/body weight ratio was measured at 7 days. The RCA/LCA peak velocity ratio at 1 day was significantly correlated with the heart weight/body weight ratio at 7 days after banding (r=0.62, p<0.005). In another 12 banded mice, serial Doppler velocity signals were obtained from the aortic banding site, the abdominal aorta (ABD) and the RCA and LCA before, 1 day after and 7 days after banding. Peak RCA velocity increased significantly after banding and both peak LCA velocity and peak ABD velocity decreased significantly. Mean velocities of RCA, LCA and ABD were unchanged before and after banding, suggesting that mice utilize peripheral arterial adaptations to maintain normal cerebral and peripheral perfusion. There was a significant positive correlation (r=0.83, p<0.001) between the RCA/LCA peak velocity ratio and peak jet velocity across the aortic banding site. Our data indicate that changes in carotid velocity after aortic banding can be used to estimate the pressure drop across the aortic band and to predict loading and resulting cardiac hypertrophy in mice. Additionally, we validated that the simplified Bernoulli equation (DeltaP=4V2) can be used to estimate the pressure drop across the aortic band in mice noninvasively. PMID- 14553806 TI - Elastography imaging of small animal oncology models: a feasibility study. AB - To test the feasibility of applying ultrasonic elastography on small animal oncology models, experiments were performed in vitro and in situ on murine mammary lesions induced exogenously by tumor cell line 66.3. In vitro studies involved three 1-week-old excised tumors embedded in a phantom block with ultrasonic properties similar to those of soft biologic tissues. In situ studies involved five mice whose bodies were embedded in pure gelatin blocks. The data were acquired from the blocks with a clinical scanner modified to have an automated compressor assembly and processed to construct the elastograms at various imaging planes within each block. The results were analyzed both qualitatively and quantitatively to assess the merits of the elastographic imaging and its limitations for in vivo serial studies of tumors in small animal oncology models. PMID- 14553807 TI - Dual lumen transducer probes for real-time 3-D interventional cardiac ultrasound. AB - We have developed dual lumen probes incorporating a forward-viewing matrix array transducer with an integrated working lumen for delivery of tools in real-time 3 D (RT3-D) interventional echocardiography. The probes are of 14 Fr and 22 Fr sizes, with 112 channel 2-D arrays operating at 5 MHz. We obtained images of cardiac anatomy and simultaneous interventional device delivery with an in vivo sheep model, including: manipulation of a 0.36-mm diameter guidewire into the coronary sinus, guidance of a transseptal puncture using a 1.2-mm diameter Brockenbrough needle, and guidance of a right ventricular biopsy using 3 Fr biopsy forceps. We have also incorporated the 22 Fr probe within a 6-mm surgical trocar to obtain apical four-chamber ultrasound (US) scans from a subcostal position. Combining the imaging catheter with a working lumen in a single device may simplify cardiac interventional procedures by allowing clinicians to easily visualize cardiac structures and simultaneously direct interventional tools in a RT3-D image. PMID- 14553809 TI - Adaptive clutter filtering for ultrasound color flow imaging. AB - In this article, we present an adaptive clutter rejection method for selecting different clutter filters in ultrasound color flow imaging. A single clutter filter is typically used to reject the clutter. Because the clutter characteristics vary in both space and time, the single clutter filter approach has difficulty in providing optimum clutter rejection in ultrasound images. To achieve more accurate velocity estimation, we have developed a method to select a clutter filter adaptively at each location in an image from a set of predefined filters. Selection criteria have been developed based on the underlying clutter characteristics and the properties of various filters (e.g., minimum-phase finite impulse response, projection-initialized infinite impulse response and polynomial regression). We have incorporated our adaptive clutter rejection method in an ultrasound system. We have found that our adaptive method can reduce the mean absolute error between the estimated and true flow velocities significantly compared with the conventional methods, in which a single clutter filter is used throughout the entire image. With in vivo abdominal data, we obtained an average gain of 5.0 dB in signal-to-clutter ratio (SCR), compared with the conventional method. These preliminary results indicate that the proposed adaptive method could improve the accuracy of flow velocity estimation in ultrasound color flow imaging through the improvement in SCR and the reduction in bias. PMID- 14553808 TI - Statistical modeling of atherosclerotic plaque in carotid B mode images--a feasibility study. AB - A feasibility study undertaken to model atherosclerotic plaque in carotid B-mode images is presented. The study is based on 33 regions-of-interest collected from arterial images obtained from four patients. A bimodal gamma distribution with five parameters is proposed to model the statistics of the pixels in the gray level images. The parameters of the distribution are evaluated for regions containing plaque using curve-fitting techniques. This bimodal distribution appears to be a reasonable fit to the statistics of the pixels. This statistical model may aid in the objective classification of arterial plaque through the use of its parameters. PMID- 14553810 TI - In vitro and in vivo ablation of porcine renal tissues using high-intensity focused ultrasound. AB - The aim of this paper is to present issues regarding the thermal ablation of porcine renal tissues in vitro and in vivo using high-intensity focused ultrasound (HIFU). Production of lesions in the cortex in vitro is consistent, whereas lesions in the medulla are created whenever there are no air spaces in the medulla. Typically, the lesion length at 2000 W/cm2 and 5-s pulse duration is around 20 mm and the corresponding width around 3 mm. Lesioning of a large volume was achieved by moving the transducer in a grid formation. Lesioning through a fat layer is possible provided that there are no air spaces between the fat and kidney interface. It was found that, above 3200 W/cm2 with 5-s pulse duration at 4 MHz, cavitation activity occurred in most of the lesions created. PMID- 14553811 TI - Effects of ultrasound on the structure and function of tumor necrosis factor alpha. AB - The objective of this study was to investigate the effects of ultrasound on the structure and function of human tumor necrosis factor-alpha (TNF-alpha) and to study whether TNF-alpha underwent a denaturation process and the molecular structure was damaged when it was irradiated by ultrasound. The samples of TNF alpha were dissolved in aqueous solution and filled into polystyrene tubes. High intensity ultrasound processor (20 kHz frequency, burst mode, 0.5 duty factor, 100-500 W total electrical power, 0-20 min total treatment time) was used during the treatment. The biologic activity of TNF-alpha was determined by its toxic activity towards TNF-alpha sensitive cell line L929 in the presence of actinomycin D. The methods of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) were used to detect the integrity of TNF-alpha molecule after it was irradiated by ultrasound. The results showed TNF alpha could keep its biological activity, instead of undergoing a denaturation process, when it is irradiated by ultrasound in the aqueous solution; at the same time, the aggregates of TNF-alpha formed by the recombinant DNA E. coli could be dissociated through the molecular vibration induced by ultrasound energy. The biologic activity of TNF-alpha was not reduced, but small quantities of TNF-alpha molecular structure were damaged during the process of sonication. These features of TNF-alpha molecule irradiated by ultrasound probably gave TNF-alpha the advantage in being used in the drug microencapsulation and provided a new drugs formulation for tumor therapy. PMID- 14553812 TI - Impact of myocardial contrast echocardiography on vascular permeability: an in vivo dose response study of delivery mode, pressure amplitude and contrast dose. AB - An in vivo rat model of myocardial contrast echocardiography (MCE) was defined and used to examine the dose range response of microvascular permeabilization and premature ventricular contractions (PVCs) with respect to method of imaging, peak rarefactional pressure amplitude (PRPA) and agent dose. A left ventricular short axis view was obtained on anesthetized rats at 1.7 MHz using a diagnostic ultrasound system with simultaneous ECG recording. Evans blue dye, a marker for microvascular leakage, and a bolus of Optison were injected i.v. Counts of PVCs were made from video tape during the 3 min of MCE. Hearts were excised 5 min after imaging and petechial hemorrhages, Evans blue colored area and Evans blue content were determined. No PVCs or microvascular leakage were seen in rats imaged without contrast agent followed by contrast agent injection without imaging. When PVCs were detected during MCE, petechial hemorrhages and Evans blue leakage were also found in the myocardium. Triggering 1:4 at end-systole produced the most PVCs per frame and most microvascular leakage, followed by end-systole 1:1, continuous scanning and end-diastole triggering 1:1. All effects increased with increasing Optison dosage in the range 25 to 500 microL kg(-1). Ultrasound PRPA was important, with apparent thresholds for PVCs at 1.0 MPa and for petechiae at 0.54 MPa. PVCs, petechial hemorrhages and microvascular leakage in the myocardium occur as a result of MCE in rats. PMID- 14553813 TI - Polyacrylamide gel as an acoustic coupling medium for focused ultrasound therapy. AB - A hydrogel acoustic coupling medium was investigated as a practical alternative to water for clinical applications of focused ultrasound (US) therapy. Material characterization and functional testing of polyacrylamide gel couplers were performed. Acoustic, bulk and thermal properties were measured. Conical couplers were designed and fabricated to fit a 3.5-MHz, spherically concave transducer for functional tests, including Schlieren imaging, power efficiency measurements and in vivo hemostasis experiments. Polyacrylamide was shown to have favorable acoustic properties that varied linearly with acrylamide concentration from 10% to 20% weight in volume. Attenuation coefficient, sound speed and impedance ranged from 0.08 to 0.14 dB/cm at 1 MHz, 1546 to 1595 m/s and 1.58 to 1.68 Mrayl, respectively. An intraoperative in vivo hemostasis experiment in a sheep model demonstrated that the gel-coupled transducer was capable of inducing hemostasis in actively bleeding splenic and hepatic incisions. The results of this study show that polyacrylamide may be a promising coupling material for focused US therapy. PMID- 14553814 TI - Ultrasound-mediated cavitation thresholds of liquid perfluorocarbon droplets in vitro. AB - This study was undertaken to measure the ultrasound (US)-mediated cavitation threshold of microdroplets as a function of its content and US parameters (frequency, amplitude and burst length). Albumin-coated droplets were prepared with perfluoropropane, perfluorohexane or perfluoromethylcyclohexane contents. The filtered suspensions were diluted to 1:1000 (v) and compared with Optison. The formulations were injected into an acoustically transparent vessel and sonicated with a single focused transducer. The frequencies employed were 0.74, 1.1, 2.18 and 3.3 MHz and the burst length and acoustic pressure were varied. The inertial cavitation threshold for each experiment was monitored through passive acoustic detection. The formation of droplet emulsion of the perfluorocarbon increased the natural boiling point of the perfluorocarbon. However, perfluorocarbon droplets having contents with higher molecular weights and boiling points did not have detectably higher inertial cavitation thresholds and, thus, the droplets do not need to be in a superheated state to be cavitated by US bursts. Therefore, higher boiling point perfluorocarbons should be investigated for this purpose and may prove to be useful for both imaging and therapy. The inertial cavitation threshold of perfluorocarbon droplets increases with frequency, and was approximately 0.7 MPa at 0.74 MHz and 1.75 MPa at 3.3 MHz. Optison, already in a gaseous state, has the lowest cavitation threshold of all formulations studied. Results show that, for the frequencies tested, there is no dependence between inertial cavitation threshold and burst lengths between 20 and 100 ms. As a conclusion, the inertial cavitation threshold of albumin-coated microdroplets of several perfluorocarbons was determined in vitro. The results indicate that the physical properties of these droplets are such that they may be useful for localized US therapies. PMID- 14553815 TI - Influence of tissue preparation on the acoustic properties of tissue sections at high frequencies. AB - The purpose of the present study was to clarify the influence of tissue preparation on the high-frequency acoustic properties by comparing the acoustic properties of the formalin-fixed, paraffin-embedded, deparaffinized sections and formalin-fixed, frozen sections for two types of fat-containing renal cancer and fat-free renal oncocytoma using a SAM. There was no significant difference for the sound speed among the clear cell, granular cell renal cancer and oncocytoma in either groups, but the attenuation constant was significantly higher for the frozen than for that of the paraffin section in fat-containing renal cancer. In fat-free oncocytoma, there was no significant difference for the attenuation constant in either group. The data suggest that the fat component, which had been eluted by paraffinization, is stored and the true acoustic properties of the tissue can be measured in frozen section. PMID- 14553816 TI - Do we need a better approach for measuring pulse-wave velocity? PMID- 14553817 TI - Do we need a better approach for measuring pulse-wave velocity? PMID- 14553818 TI - The cardiac cocaine connection. PMID- 14553819 TI - Amlodipine and endothelial nitric oxide synthase activity. PMID- 14553820 TI - Of mice and men, rats and atherosclerosis. PMID- 14553821 TI - Matrix metalloproteinases: a review of their structure and role in acute coronary syndrome. AB - OBJECTIVE: To summarize the structure, function, and regulation of matrix metalloproteinases (MMPs) and to review the literature to date on their potential role in the pathogenesis of acute coronary syndromes. METHODS: A recursive strategy starting with a Medline Search for primary articles in the last decade, followed by identification of additional articles of interest among the cited literature in the primary articles, followed by identification of additional articles of interest cited in the secondary articles. RESULTS: MMPs play a central role in many fundamental processes in human health and disease. In vitro evidence suggests that MMP activity may facilitate atherosclerosis, plaque destabilization, and platelet aggregation. Limited evidence from clinical studies supports a role of MMPs in the development of acute coronary syndromes. CONCLUSIONS: MMP activity likely contributes to the development of acute coronary syndromes and may be an important therapeutic target for future drug development. PMID- 14553822 TI - The emerging role of asymmetric dimethylarginine as a novel cardiovascular risk factor. AB - There is abundant evidence that the endothelium plays a crucial role in the maintenance of vascular tone and structure. One of the major endothelium-derived vasoactive mediators is nitric oxide (NO). Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of NO synthase. ADMA inhibits vascular NO production at concentrations found in pathophysiological conditions (i.e., 3-15 micromol/l); ADMA also causes local vasoconstriction when it is infused intraarterially. The biochemical and physiological pathways related to ADMA are now well understood: dimethylarginines are the result of the degradation of methylated proteins; the methyl group is derived from S-adenosylmethionine. Both ADMA and its regioisomer, SDMA, are eliminated from the body by renal excretion, whereas only ADMA, but not SDMA, is metabolized via hydrolytic degradation to citrulline and dimethylamine by the enzyme dimethylarginine dimethylaminohydrolase (DDAH). DDAH activity and/or expression may therefore contribute to the pathogenesis of endothelial dysfunction in various diseases. ADMA is increased in the plasma of humans with hypercholesterolemia, atherosclerosis, hypertension, chronic renal failure, and chronic heart failure. Increased ADMA levels are associated with reduced NO synthesis as assessed by impaired endothelium-dependent vasodilation. In several prospective and cross-sectional studies, ADMA evolved as a marker of cardiovascular risk. With our increasing knowledge of the role of ADMA in the pathogenesis of cardiovascular disease, ADMA is becoming a goal for pharmacotherapeutic intervention. Among other treatments, the administration of L arginine has been shown to improve endothelium-dependent vascular function in subjects with high ADMA levels. PMID- 14553823 TI - Role of reactive oxygen species in cocaine-induced cardiac dysfunction. AB - OBJECTIVE: Contractility alterations and LV hypertrophy after chronic cocaine administration have been shown to be accompanied by an increase in oxidative stress. This study was carried out to investigate whether the production of reactive oxygen species is an early event of primary importance in cocaine induced myocardial injury or simply occurs as a consequence of the ventricular dysfunction itself. METHODS AND RESULTS: After 2 days of cocaine administration to rats, no differences were observed in echocardiographic parameters between the cocaine-treated group and the control group. However, an increase in oxidative stress in the myocardium was indicated by an increase in lipid peroxidation (+35%, cocaine vs. control), an increase in antioxidant enzymes (catalase +110%, glutathione peroxidase +40% and superoxide dismutase +38%) and of NADPH-driven superoxide production (assessed by chemiluminescence). Furthermore, higher gp91phox and p22phox mRNA expression, measured by quantitative real-time RT-PCR, was found in the cocaine group. On day 8, cocaine administration induced a cardiac dysfunction, characterized by a decrease in cardiac index (-30%, cocaine vs. controls) and left ventricular (LV) fractional shortening (-23%, cocaine vs. controls). This LV dysfunction was prevented by antioxidant treatment (100 mg/kg/day vitamin C and 100 U/kg/day vitamin E). Moreover, in these animals, antioxidant treatment decreased lipid peroxides and decreased the activity of NADPH oxidase, associated with the downregulation of gp91phox. CONCLUSION: These data indicate that cocaine administration induces early NADPH-driven O2-. release which may play an important role in the development and progression of the LV dysfunction observed after chronic cocaine abuse. PMID- 14553824 TI - Amlodipine activates the endothelial nitric oxide synthase by altering phosphorylation on Ser1177 and Thr495. AB - OBJECTIVE: The Ca2+ antagonist amlodipine increases the generation of nitric oxide (NO) from native and cultured endothelial cells. The aim of this investigation was to determine whether or not the activation of the endothelial NO synthase (eNOS) by this Ca2+ antagonist is related to alterations in eNOS phosphorylation. METHODS AND RESULTS: In isolated, pre-contracted, endothelium intact porcine coronary arteries, amlodipine elicited an NO-mediated relaxation and a leftward shift in the concentration-relaxation curve to bradykinin. Moreover, the Ca2+ antagonist increased the generation of NO from native endothelial cells, as detected by electron spin resonance spectroscopy and stimulated an 8-fold increase in cyclic GMP levels in cultured endothelial cells. In unstimulated endothelial cells, eNOS was not phosphorylated on Ser1177 but was phosphorylated on Thr495. Amlodipine elicited the phosphorylation of Ser1177 and attenuated Thr495 phosphorylation, with a time course similar to that of eNOS activation. The amlodipine-induced relaxation of porcine coronary arteries was attenuated by the B2 kinin receptor antagonist, icatibant, but this antagonist did not affect amlodipine-induced changes in eNOS phosphorylation in cultured endothelial cells. Moreover, amlodipine elicited the NO-mediated relaxation of rat aortic rings which do not express the B2 receptor. Amlodipine time dependently attenuated the phosphorylation of protein kinase C (PKC) in endothelial cells, with a time course similar to the changes in eNOS phosphorylation, and prevented the phorbol-12-myristate-13-acetate-induced activation of PKC. The PKC inhibitor, Ro 31-8220, also elicited the phosphorylation of Ser1177 and the dephosphorylation of Thr495 in cultured cells and induced a leftward shift in the concentration-relaxation curve to bradykinin in rings of porcine coronary artery. CONCLUSION: The Ca2+ antagonist, amlodipine, enhances endothelial NO generation by inducing changes in the phosphorylation of eNOS. Although the activation of eNOS was related to the activation of the B2 kinin receptor in the porcine coronary artery, a B2 receptor-independent mechanism involving the inhibition of PKC appears to account for the effects observed in the rat aorta as well as in cultured endothelial cells. PMID- 14553825 TI - Paradoxical reduction of atherosclerosis in apoE-deficient mice with obesity related type 2 diabetes. AB - OBJECTIVE: The effect of obesity and insulin resistance on the development of atherosclerosis was evaluated in apoE-deficient (ApoE(-/-)) mice. A previously described obesity model, in which the hypothalamic satiety center can be destroyed by a single gold thioglucose (GTG) injection, was used. To evaluate the effect of starvation on atherosclerosis ApoE(-/-) mice were food-restricted with 25% less chow than ad libitum-fed control mice. METHODS: Sixty-eight ApoE(-/-) mice were allocated into a control group (n=20), a GTG-injected group (n=28), and a food-restricted group (n=20). The control and food-restricted mice were injected with saline instead of GTG. The control and GTG-injected mice had free access to food, and all mice had free access to water during the study period. RESULTS: After 4 months, the GTG-injected mice were significantly overweight (mean body weight (g): 33 +/- 2.11 vs. 23 +/- 0.24 and 17 +/- 0.31 in control and food-restricted mice, respectively), obese, hypertriglyceridemic, insulin resistant, hyperinsulinemic (mean plasma insulin (ng/ml): 2.45 and 0.43 in obese and control mice, respectively), and hyperglycemic (mean plasma glucose (mmol/l): 11.03 and 7.80 in obese and control mice, respectively). Unexpectedly, these obese and diabetic mice developed significantly less atherosclerosis compared with lean non-diabetic control mice. Food-restricted mice also developed less atherosclerosis compared to control mice. CONCLUSIONS: These findings may question the usefulness of mouse models in studying the relation of obesity related type 2 diabetes to atherosclerosis and also the relevance of results obtained in apoE(-/-) mice with reduced weight gain during intervention. PMID- 14553826 TI - Cholinergic atrial fibrillation: I(K,ACh) gradients determine unequal left/right atrial frequencies and rotor dynamics. AB - OBJECTIVE: We tested the hypothesis that left atrial (LA) myocytes are more sensitive to acetylcholine (ACh) than right atrial (RA) myocytes, which results in a greater dose-dependent increase in LA than RA rotor frequency, increased LA to-RA frequency gradient and increased incidence of wavelet formation during atrial fibrillation (AF). METHODS AND RESULTS: AF was induced in seven Langendorff-perfused sheep hearts in the presence of ACh (0.1-4.0 microM) and studied using optical mapping and bipolar recordings. Dominant frequencies (DFs) were determined in optical and electrical signals and phase movies were used to identify rotors and quantify their dynamics. DFs in both atria increased monotonically with ACh concentration until saturation, but the LA frequency predominated at all concentrations. Rotors were also seen more often in the LA, and although their life span decreased, their frequency and number of rotations increased. Patch-clamp studies demonstrated that ACh-activated potassium current (I(K,ACh)) density was greater in LA than RA sheep myocytes. Additionally, ribonuclease protection assay demonstrated that Kir3.4 and Kir3.1 mRNAs were more abundant in LA than in RA. CONCLUSIONS: A greater abundance of Kir3.x channels and higher I(K,ACh) density in LA than RA myocytes result in greater ACh-induced speeding-up of rotors in the LA than in the RA, which explains the ACh dose dependent changes in overall AF frequency and wavelet formation. PMID- 14553827 TI - Cellular engineering of ventricular adult rat cardiomyocytes. AB - OBJECTIVE: Preparation of viable cultured adult cardiomyocytes (vARCs) is a prerequisite for cell-based transplantation and tissue engineering. Ectopic gene expression is important in this context. Here, we present an in vitro cell replating strategy using Accutase for cultured vARCs, allowing ectopic gene expression. METHODS: Cultured vARCs from 6- to 8-week-old rats were used. Transfections with EGFP (enhanced green fluorescent protein) constructs, Mlc-3f EGFP or alpha-actinin-EGFP were performed using adenovirus-enhanced transferrin mediated infection (AVET). Accutase (PAA Laboratories, Linz, Austria) was used for the detachment of cultured cells. Immunohistochemical analysis, together with confocal laser microscopy was used for structural analysis of the cells. RESULTS: Cultured vARCs could be detached with a high yield (40 to 60%) from primary cultures using Accutase. The cultivation period plays an important role in the yield of viable cells. Resultant replated vARCs (rep-vARCs) rapidly (1-2 h) acquired a rounded up shape without degradation of their contractile apparatus, which is in contrast to the rod-shaped freshly isolated vARCs (fi-vARCs). The detached cells survived passage through a narrow syringe needle. After seeding, detached cells rapidly attached to various substrates, increased their content of the contractile apparatus, and formed cell-cell contacts within 3 days after reseeding. The detached cells survived passage through a narrow syringe needle. The high recovery of cells after replating enabled the use of the AVET system for gene delivery. AVET is free of infectious particles and does not lead to expression of viral proteins. Transfection of vARCs prior to detachment had a small effect on cell recovery and ectopically synthesized proteins were properly localized after replating. CONCLUSIONS: Detachment of cultured vARCs using Accutase is well compatible with ectopic gene expression and yields a viable transgenic population of vARCs that eventually may be suitable as transgenic cardiomyocyte grafts. PMID- 14553828 TI - Tamoxifen and ICI 182,780 negatively influenced cardiac cell growth via an estrogen receptor-independent mechanism. AB - BACKGROUND: Recent studies have demonstrated that selective estrogen receptor modulators (SERMs) reduced identifiable risk factors implicated in cardiovascular disease. Despite this observation, the direct effect of SERMs on cardiac cell growth remains unexplored. METHODS: Neonatal rat cardiac myocytes (CM) and fibroblasts (CF) were exposed to either the partial estrogen receptor agonist/antagonist 4-OH tamoxifen (e.g., SERM) or the pure estrogen receptor antagonist ICI 182,780 and the effect on DNA synthesis, cell cycle protein expression and extracellular signal-regulated kinase (ERK1/2) phosphorylation were assessed. RESULTS: The treatment of CM and CF with either 4-OH tamoxifen or ICI 182,780 decreased DNA synthesis in the absence of apoptosis via an estrogen receptor-independent pathway. In CM and CF, 4-OH tamoxifen and ICI 182,780 treatment reduced proliferating cell nuclear antigen protein expression and concomitantly increased p27Kip1. 4-OH Tamoxifen and ICI 182,780 treatment increased ERK1/2 phosphorylation in CM and CF, and ERK1/2 kinase (MEK)-dependent inhibition of ERK1/2 activation attenuated ICI 182,780-mediated suppression of DNA synthesis. CONCLUSION: These data are the first to describe cardiac cells as novel targets of SERMs and ICI 182,780, and highlight the role of the ERK1/2 pathway in the suppression of DNA synthesis. PMID- 14553829 TI - Inhibition of p38 MAPK decreases myocardial TNF-alpha expression and improves myocardial function and survival in endotoxemia. AB - OBJECTIVES: The role of p38 mitogen-activated protein kinase (MAPK) activation in lipopolysaccharide (LPS)-induced myocardial dysfunction has not been clearly defined. Our aim was to investigate the contribution of p38 MAPK in myocardial tumor necrosis factor-alpha (TNF-alpha) expression, cardiac function and survival during acute endotoxemia in mice. METHODS: Acute endotoxemia was induced by LPS (10 mg/kg, i.p.) in mice. Two hours after LPS treatment, left ventricular (LV) function was assessed. Phosphorylation of p38 MAPK was measured by Western blotting. TNF-alpha mRNA and protein levels were determined by semi-quantitative reverse-transcriptase polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. RESULTS: LPS rapidly increased phosphorylation of p38 MAPK, followed by TNF-alpha mRNA expression and protein expression in the LV myocardium. Pre-treatment of the p38 MAPK inhibitor SB202190 (2 mg/kg, i.p.) decreased TNF-alpha mRNA and protein by 65 and 36%, respectively (P<0.05). Immunohistochemical staining confirmed that cardiomyocytes were the major source of TNF-alpha production in the myocardium and blocking p38 MAPK activation inhibited TNF-alpha expression in response to LPS. Pre-treatment of SB202190 or a TNF-alpha antagonist etanercept (2 mg/kg, i.p) significantly reversed LPS-induced LV depression (P<0.05). LPS (20 mg/kg, i.p.) induced 94% mortality in mice within 72 h and pre-treatment with SB202190 and etanercept decreased LPS-induced mortality to 65 and 40%, respectively (P<0.01). CONCLUSION: p38 MAPK activation represents an important mechanism leading to myocardial TNF-alpha production and cardiac dysfunction during acute endotoxemia in mice. Our data suggest that p38 MAPK is a potential therapeutic target of endotoxemia. PMID- 14553830 TI - Delayed preconditioning in cardiac myocytes with respect to development of a proinflammatory phenotype: role of SOD and NOS. AB - OBJECTIVE: Both superoxide dismutase (SOD) and nitric oxide synthase (NOS) have been implicated in delayed preconditioning (DP) to ischemia/reperfusion (I/R) in the heart. We used isolated cardiac myocytes to test the hypothesis that SOD and NOS may interact in the development of DP. METHODS: Mouse neonatal cardiac myocytes were challenged with anoxia/reoxygenation (A/R; an in vitro counterpart to I/R) and normoxia/normoxia (N/N) served as the control. Two indices of inflammation were measured: oxidant stress (DHR oxidation) and polymorphonuclear leukocyte (PMN) transendothelial migration (cell culture inserts). The role of SOD was assessed using an antisense approach and the role of NOS was assessed using iNOS and eNOS deficient myocytes. RESULTS: Cardiac myocytes exposed to A/R (1) produced more oxidants (intracellular fluorescence emission from 2.0 +/- 0.1 for N/N to 3.0 +/- 0.3 for A/R; P<0.05) and (2) promoted PMN migration (% migration from 8.4 +/- 0.9 for N/N to 14.1 +/- 1.1 for A/R; P<0.05). DP occurred if the myocytes were pretreated with an A/R challenge 24 h earlier. That is, these A/R-induced responses were significantly reduced (fluorescence emission 1.9 +/- 0.1 and % migration 8.4 +/- 0.7; P<0.05 as compared to A/R with no pretreatment). Myocyte Mn-SOD, but not Cu/Zn-SOD, activity increased 24 h after the initial A/R challenge. A Mn-SOD antisense oligonucleotide prevented the development of DP. DP occurred in iNOS, but not eNOS, deficient myocytes. A/R increased mRNA for eNOS, but not iNOS, in wild-type myocytes. A/R increased Mn SOD protein in both iNOS and eNOS deficient myocytes. However, Mn-SOD activity increased only in iNOS deficient myocytes. CONCLUSIONS: Collectively, these findings suggest that Mn-SOD and eNOS may act in concert in the development of DP in cardiac myocytes. PMID- 14553831 TI - Cardiotonic drugs differentially alter cytosolic [Ca2+] to left ventricular relationships before and after ischemia in isolated guinea pig hearts. AB - OBJECTIVE: Cardiotonic agents may differentially alter indices of the cytosolic [Ca2+]/left ventricular pressure (LVP) relationship when given before and after ischemia. We measured and calculated systolic-diastolic [Ca2+], systolic diastolic LVP, velocity ratios (VRs) d[Ca2+]/dtmax to dLVP/dtmax (VRmax), d[Ca2+]/dtmin to dLVP/dtmin (VRmin), and area ratio (AR, area Ca2+]/area LVP per beat) before and after 30 min global ischemia in guinea pig hearts. METHODS: Hearts were perfused with levosimendan, dobutamine, dopamine, or digoxin. Ca2+ transients were recorded by indo-1 fluorescence via a fiber optic probe placed on the LV free wall. [Ca2+]/LVP loops were acquired by plotting LVP time as a function of [Ca2+] at multiple time points during the cardiac cycle. RESULTS: Ischemia reperfusion increased [Ca2+] and decreased contractility and relaxation and produced a flatter and broader [Ca2+]/LVP loop. All drugs shifted the [Ca2+]/LVP loop rightward and upward when given before and after ischemia. Dobutamine increased [Ca2+] and contractility more than other drugs. Digoxin increased [Ca2+] the least but increased contractility similar to dopamine and levosimendan. Before ischemia dopamine and digoxin both decreased VRmax and VRmin, whereas dobutamine increased VRmin, but not VRmax, and levosimendan had no effect on VR. VRmax and VRmin were markedly elevated after ischemia, but again decreased with dopamine and digoxin; dobutamine again increased VRmin, but not VRmax, and levosimendan decreased both VRmax and VRmin. Before ischemia dopamine and digoxin both decreased AR, dobutamine increased AR, and levosimendan had no effect; after ischemia AR was markedly elevated but dopamine and digoxin decreased AR, dobutamine increased AR, and levosimendan decreased AR. CONCLUSION: Although each drug enhanced contractility and relaxation both before and after ischemia by increasing cytosolic [Ca2+] and Ca2+ flux, dopamine and digoxin improved, and dobutamine worsened responsiveness to Ca2+, i.e., velocity ratio and area ratio, whereas levosimendan had no net effect before ischemia but improved responsiveness after ischemia. PMID- 14553832 TI - Post-ischemic myocardial fibrosis occurs independent of hemodynamic changes. AB - OBJECTIVES: Myocardial fibrosis is a major component of ventricular remodeling after large myocardial infarction (MI). The present study tests the hypothesis that post-ischemic myocardial fibrosis can occur independent of hemodynamic changes. METHODS: A mouse model of distal left coronary artery ligation was established to induce a small infarct (less than 15% of the left ventricle) in order to avoid significant mechanical overload after permanent myocardial ischemia. Left heart catheterization was performed to evaluate the post-infarct hemodynamics. Tissues from both ischemic and non-ischemic myocardium were examined for mRNA and protein expression at 24, 72 h and 7 days after ligation. RESULTS: Heart/body weight ratio after ligation was increased by approximately 10% over sham control although there is no statistically significant difference in hemodynamic parameters between the two groups. Non-ischemic myocardium distant from the infarct site showed molecular evidence of myocardial fibrosis 72 h and 7 days after ligation. There was marked up-regulation of mRNAs for extracellular matrix (ECM) proteins and their cross-linking enzyme, such as collagens type I, III and VI, and lysyl oxidase. Immunohistochemical study confirmed that the expression of these ECM proteins was significantly increased in the non-ischemic myocardium after 7 days. TGF-beta1 was up-regulated after 72 h in both ischemic and non-ischemic myocardium. CONCLUSIONS: Molecular and histopathological findings demonstrate that abnormal myocardial fibrosis can be induced by a small infarct independent of secondary hemodynamic changes. PMID- 14553833 TI - PKC/Raf/MEK/ERK signaling pathway modulates native-LDL-induced E2F-1 gene expression and endothelial cell proliferation. AB - BACKGROUND AND OBJECTIVES: The interactions of low-density lipoprotein (LDL) with the endothelium are thought to play a major role in the development of atherosclerosis. Due to this reason, the molecular sequelae of events resulting from native LDL (N-LDL) interaction with human endothelial cells (HECs) are largely under investigation. METHODS AND RESULTS: Here, we report that the exposure of serum-free HECs to different concentrations of N-LDL-cholesterol (LDL chol) elicited a time- and dose-dependent induction of DNA synthesis. The exposure of serum-free HECs to N-LDL was able to elicit a time- and dose dependent increase of protein kinase C (PKC) activity that, along with the activation of the Raf/mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) signaling pathway, leads to an increase in E2F-1 gene expression. In addition, the treatment of HECs with N-LDL was also able to induce both E2F-1 gene transcription and protein expression. These N-LDL-aroused responses were dramatically counteracted by PKC inhibition or down regulation. Similarly to what observed for Raf/MEK/ERK activation and E2F-1 gene expression, the inhibition of PKC as well as its down regulation, significantly lowered the DNA synthesis induced by N-LDL in serum-free HECs. CONCLUSIONS: These results suggest that the activation of PKC/Raf/MEK/ERK-mediated events controlling E2F-1 gene expression by N-LDL may represent an important mechanism in the regulation of HECs proliferation during normal and pathological processes. PMID- 14553834 TI - 8-isoprostane increases expression of interleukin-8 in human macrophages through activation of mitogen-activated protein kinases. AB - BACKGROUND AND OBJECTIVES: 8-isoprostane is a marker of oxidative stress in vivo and increased plasma and urine levels are found in patients with vascular disease and in atherosclerotic plaques. Inflammatory chemokines such as interleukin (IL) 8 seem to play an important pathogenic role in atherogenesis. We therefore investigated the effects of 8-isoprostane on the expression of inflammatory chemokines with consciousness on IL-8 (mRNA and protein) in human macrophages. In addition, we studied the involvement of mitogen-activated protein kinases (ERK 1/2 and p38 MAPK) and nuclear factor-kappaB (NF-kappaB) in this process. METHODS AND RESULTS: 8-isoprostane (10 microM) induced IL-8 expression (mRNA and protein), measured by real-time quantitative RT-PCR and enzyme immunoassay, respectively, in both THP-1 macrophages and human monocyte-derived macrophages. Moreover, 8-isoprostane increased mRNA expression of macrophage inflammatory protein-1alpha as determined by RNase protection assay. In this process, 8 isoprostane induced the activation of two major MAP-kinases; ERK 1/2 and p38 MAPK. Furthermore, the ERK 1/2 inhibitor, PD98059, and the p38 MAPK inhibitor, SB203580, markedly reduced 8-isoprostane-induced IL-8 expression (mRNA and protein), while inhibition of NF-kappaB activation and translocation had no significant effect on IL-8 expression. CONCLUSIONS: We show that 8-isoprostane increases IL-8 expression in human macrophages involving both ERK 1/2 and p38 MAPK, but not NF-kappaB signaling pathway. These findings further support a link between oxidative stress/lipid peroxidation and inflammation in human macrophages and suggest a role for 8-isoprostane in this process. This 8-isoprostane-induced chemokine expression might be involved in the pathogenesis of atherosclerosis as well as other inflammatory disorders. PMID- 14553835 TI - The effects of dietary fatty acid supplementation on endothelial function and vascular tone in healthy subjects. AB - OBJECTIVE: Evaluation of the effects of supplementation of n-3 and n-6 fatty acids on vascular tone and endothelial function in healthy men and women aged 40 to 65 years. METHODS: In a double-blind, randomised, placebo controlled study, 173 healthy volunteers took one of six oil supplements for 8 months. Supplements were placebo, oleic acid rich sunflower oil, evening primrose oil, soya bean oil, tuna fish oil, and tuna/evening primrose oil mix. Endothelium-dependent and independent vascular responses were measured in the forearm skin using laser Doppler imaging following iontophoretic applications of acetylcholine and sodium nitroprusside, respectively. RESULTS: Acetylcholine, but not sodium nitroprusside responses were significantly improved after tuna oil supplementation (P=0.02). Additionally, there were significant positive correlations between acetylcholine responses and n-3 fatty acid levels in the plasma and erythrocyte membrane phospholipids after tuna oil supplementation. No significant changes in vascular response were seen after supplementation with any of the other oils. CONCLUSIONS: Fish oil supplementation has a beneficial effect on endothelial function, even in normal healthy subjects. Modification of the diet by an increase of 6% in eicosapentaenoic acid and 27% in docosahexaenoic acid (equivalent to eating oily fish 2-3 times/week) might have significant beneficial effects on cardiovascular function and health. PMID- 14553836 TI - Reduction of endothelial NOS and bradykinin-induced extravasation of macromolecules in skeletal muscle of the fructose-fed rat model. AB - OBJECTIVE: Reduced capillary permeability of the skeletal muscle vascular bed has been suggested to play a role in fructose-fed rats, corroborating a long held view that insulin resistance might partially be explained by the lack of access of insulin and glucose to its target organs, mainly skeletal muscles. The goal of this study was to explore mechanisms underlying this vascular abnormality, and more specifically the role of bradykinin and nitric oxide (NO) on skeletal muscle microcirculation and the extravasation of macromolecules. METHODS: For that purpose, Sprague-Dawley rats were fed with either a fructose-enriched (F) or a normal chow (N) diet and extravasation of macromolecules was assessed at 4 weeks by measuring in vivo the extravasation of Evans Blue (EB) dye in the quadriceps muscles of both groups after the intravenous injection of the potent vasodilator bradykinin (150 microg/kg). RESULTS: As expected, fructose-fed rats had less extravasation of EB in skeletal muscle in the basal state as compared to controls (F 17.6 +/- 4.4 vs. N 43.6 +/- 6.9 microg/g dry tissue; P<0.01). In response to bradykinin, the EB dye extravasation in skeletal muscle was 89.4% higher in rats fed the normal chow diet compared to the basal state (P<0.03). In contrast, no significant increase in vasopermeability was observed in fructose-fed animals acutely injected with BK (17.6 +/- 4.4 microg/g in the basal state versus 24.6 +/ 3.1 microg/g after the injection of BK; P=NS). To distinguish a functional from an anatomical/structural defect, hematoxylin-eosin sections as well as electron micrographs of skeletal muscle microvessels were examined in both groups of animals: no obvious abnormalities were found. However, in homogenates of skeletal muscles (quadriceps) of fructose-fed rats, there was a marked reduction of NO synthase (NOS) activity (-33.8%; P<0.001) as well as endothelial NOS immunoreactive mass (-23.4%; P<0.04) as compared to control animals. CONCLUSION: There is unresponsiveness of the skeletal muscle capillary bed to bradykinin in insulin-resistant animals most probably due to a reduction in endothelial NOS (activity and mass). Our results indicate a functional defect possibly involving responsiveness of the precapillary resistance and/or the endothelial barrier to bradykinin in skeletal muscles. Since insulin must cross the endothelial monolayer to reach its target cells on the abluminal side, it is suggested that reduced endothelial NOS and consequent reduced extravasation of macromolecules could exacerbate insulin resistance in skeletal muscles and hypertension in the fructose-fed rat. PMID- 14553838 TI - Differential localisation of the renin-angiotensin system in de-novo lesions and in-stent restenotic lesions in in-vivo human coronary arteries. AB - OBJECTIVE: Different components of the renin-angiotensin system (RAS) have been demonstrated in atherosclerotic plaques. However, the involvement of the RAS in in-stent restenosis is not clear. We studied the differential immunolocalisation of angiotensin converting enzyme (ACE) and the angiotensin II type 1 (AT1) receptor in de-novo stenotic lesions and in-stent restenotic lesions in human coronary arteries. METHODS: Using a pullback atherectomy catheter, biopsies from de-novo coronary lesions (n=19) and in-stent restenotic lesions (n=19) were obtained. The biopsies were immunostained for vascular smooth muscle cells (VSMCs), macrophages, ACE and the AT1 receptor. RESULTS: In biopsies from de-novo stenotic lesions ACE-positive macrophages were more numerous than in in-stent restenotic lesions (P=0.002). Moreover, in the latter lesions, ACE-positive macrophages decreased when the time interval of stent implantation was longer. On the other hand, in-stent restenotic lesions contained predominantly young VSMCs, which abundantly expressed AT1 receptors. CONCLUSIONS: Lesional ACE expression is not a prominent feature of in-stent restenotic lesions. In contrast, AT1 receptors are abundantly expressed on young VSMCs. In de-novo lesions ACE and AT1 receptors were found on macrophages and VSMCs, which were present in all specimens. PMID- 14553837 TI - Vascular endothelial growth factor-D expression in human atherosclerotic lesions. AB - OBJECTIVE: Vascular endothelial growth factor-D (VEGF-D) is a recently characterized member of the VEGF family, but its expression in atherosclerotic lesions remains unknown. We studied the expression of VEGF-D and its receptors (VEGFR-2 and VEGFR-3) in normal and atherosclerotic human arteries, and compared that to the expression pattern of VEGF-A. METHODS: Human arterial samples (n=39) obtained from amputation operations and fast autopsies were classified according to the stage of atherosclerosis and studied by immunohistochemistry. The results were confirmed by in situ hybridization and RT-PCR. RESULTS: We found that while VEGF-A expression increased during atherogenesis, VEGF-D expression remained relatively stable only decreasing in complicated lesions. In normal arteries and in early lesions VEGF-D was mainly expressed in smooth muscle cells, whereas in complicated atherosclerotic lesions the expression was most prominent in macrophages and also colocalized with plaque neovascularization. By comparing the staining profiles of different antibodies, we found that proteolytic processing of VEGF-D was efficient in the vessel wall. VEGFR-2, but not VEGFR-3, was expressed in the vessel wall at every stage of atherosclerosis. CONCLUSIONS: Our results suggest that in large arteries VEGF-D is mainly expressed in smooth muscle cells and that it may have a role in the maintenance of vascular homeostasis. However, in complicated lesions it was also expressed in macrophages and may contribute to plaque neovascularization. The constitutive expression of VEGFR-2 in arteries suggests that it may be one of the principal mediators of the VEGF-D effects in large arteries. PMID- 14553840 TI - Enhanced hindlimb collateralization induced by acidic fibroblast growth factor is dependent upon femoral artery extraction. AB - Recent investigations have established the feasibility of using exogenously delivered angiogenic growth factors to increase collateral artery development in animal models of myocardial and hindlimb ischemia. OBJECTIVE: Our aim was to evaluate the ability of a stabilized form of acidic fibroblast growth factor (aFGF-S117) to stimulate collateralization and arteriogenesis in the rabbit hindlimb following the surgical induction of ischemia by femoral artery extraction. A secondary objective was to examine angiogenic and arteriogenic effects of aFGF-S117 in the absence of a peripheral blood flow deficit. METHODS AND RESULTS: Five days after femoral artery removal, aFGF-S117 (1, 3, or 30 microg/kg) was intramuscularly delivered into the hindlimb, three times per week for 2 consecutive weeks. End-point measurements performed on day 20 found that hindlimb reserve blood flow was significantly improved in rabbits that received 3 or 30 microg/kg of aFGF-S117, with no difference in efficacy between these two doses. These hemodynamic results were supported by angiographic evidence showing enhanced density of collateral vessels in the medial thigh region and histological findings of increased capillary density within the gastrocnemius muscle from rabbits treated with aFGF-S117. When an efficacious dose of 3 microg/kg of aFGF-S117 was administered to sham-operated rabbits with intact femoral arteries, there was no change in any of the blood flow, angiographic or histological parameters measured. CONCLUSIONS: These findings demonstrate that a stabilized form of aFGF stimulated the development of functional collateral arteries in the rabbit hindlimb, an effect which was dependent upon removal of the femoral artery. These results suggest that aFGF-S117 may have therapeutic potential for the treatment of arterial occlusive disorders. PMID- 14553839 TI - Oxidative stress in the pathogenesis of thoracic aortic aneurysm: protective role of statin and angiotensin II type 1 receptor blocker. AB - OBJECTIVE: The pathogenesis of thoracic aortic aneurysms (TAA) is still unclear. A recent investigation indicated that angiotensin II, a potent activator of NADH/NADPH oxidase, plays an important role in aneurysmal formation. We investigated the potential role of p22phox-based NADH/NADPH oxidase in the pathogenesis of TAA. METHODS: Human thoracic aneurysmal (n=40) and non-aneurysmal (control, n=39) aortic sections were examined, and the localization of p22phox, an essential component of the oxidase, and its expressional differences were investigated by immunohistochemistry and Western blot. In situ reactive oxygen species (ROS) generation was examined by the dihydroethidium method, and the impact of medical treatment on p22phox expression was investigated by multiple regression analysis. RESULTS: In situ production of ROS and the expression of p22phox increased markedly in TAA throughout the wall, and Western blot confirmed the enhanced expression of p22phox. The expression was more intense in the regions where monocytes/macrophages accumulated. In these inflammatory regions, numerous chymase-positive mast cells and angiotensin converting enzyme-positive macrophages were present. Their localization closely overlapped the in situ activity of matrix metalloproteinase and the expression of p22phox. Multiple regression analysis revealed that medical treatment with statin and angiotensin II type 1 receptor blocker (ARB) suppressed p22phox expression in TAA. CONCLUSION: Our findings indicate the role of p22phox-based NADH/NADPH oxidase and the local renin-angiotensin system in the pathogenesis of TAA. Statin and ARB might have inhibitory effects on the formation of aneurysms via the suppression of NADH/NADPH oxidase. PMID- 14553841 TI - The nouns that define us. PMID- 14553842 TI - Who are we and what do we think? AB - BACKGROUND: The American Society of Breast Surgeons was founded on the principal of professional and practical development of its members. The Society undertook a survey of its members to identify issues and concerns and to guide future action. METHODS: The survey was prepared by the membership committee and a website was created for responses and analysis. The survey was posted on the website and mailed to 1530 members. Overall there was a 31% response. RESULTS: Demographic data regarding practice patterns, technology utilization, and current problems showed a diverse Society with varying needs and problems. CONCLUSIONS: The Society will pursue professional development in ultrasonography training, coding and reimbursement, and emerging technologies. The Society is committed to providing regional training programs, certification, and professional interaction. The Society will continue to look to the members for feedback and guidance. PMID- 14553843 TI - Interpretive disparity among pathologists in breast sentinel lymph node evaluation. AB - BACKGROUND: Immunohistochemical staining on breast sentinel lymph nodes (SLN) is controversial. METHODS: Twenty-five SLN cases were reviewed by 10 pathologists (three academic, seven private) including 5 negative by both hematoxylin and eosin (H&E) and immunohistochemistry, 11 micrometastases (<2 mm) negative by H&E but positive by immunohistochemistry, and 8 micrometastases and 1 macrometastasis (>2 mm) positive for both H&E and immunohistochemistry. Answers included "positive," "negative," and "indeterminate" for each slide. RESULTS: The mean number of incorrect responses was 6.6 for immunohistochemistry and 5 for H&E. Twelve percent of cases were correct by all 10 pathologists; 80% of positive IHC cases had at least one pathologist score it incorrectly. As tumor cells decrease in number, incorrect responses increase. When tumor cells numbered less than 10, more than 30% of pathologists answered incorrectly. CONCLUSIONS: As tumor cells decrease in number pathologists' ability to recognize them decreases. We propose adding "indeterminate" to "positive" and "negative" when tumor cells number less than 10. PMID- 14553844 TI - Success of sentinel lymph node mapping after breast cancer ablation with focused microwave phased array thermotherapy. AB - BACKGROUND: Breast cancer tumor ablation as part of a multimodality approach in the treatment of breast cancer is the subject of recent interest. This study was conducted to determine if the ability to perform sentinel node biopsy was impaired after thermal-induced ablation of breast cancer. METHODS: We studied patients who had sentinel node biopsy after preoperative focused microwave phased array for breast cancer ablation. RESULTS: Twenty-one patients with T1-T2 breast cancer and clinically negative axilla underwent wide local excision and sentinel node biopsy guided by blue dye and sulfur colloid. Surgery was done an average of 17 days after microwave ablation. Fifteen of 22 patients (68%) had histologic evidence of tumor necrosis. Sentinel lymph node mapping was successful in 19 of 21 patients (91%). Axillary metastases were detected in 42% of cases. CONCLUSIONS: This study documents successful sentinel lymph node mapping for patients treated with antecedent local tumor ablation using focused microwave phased array ablation. PMID- 14553845 TI - The clinical relevance of positive sentinel nodes only versus positive nonsentinel lymph nodes in breast cancer patients. AB - BACKGROUND: The purpose of this study was to determine the difference in clinical outcomes for patients with histologically positive sentinel lymph nodes (SLN+) compared with patients with histologically positive nonsentinel second echelon lymph nodes (NSLN+). METHODS: Eight hundred thirteen node positive patients from a prospectively accrued database of 3200 patients who underwent sentinel node mapping were evaluated. In all, 506 of the 813 patients (62%) were SLN+ only and 307 of the 813 patients (38%) were SLN+ plus at least one NSLN+. Patients' overall survival and disease-free survival were obtained and statistical analyses performed comparing the two groups. RESULTS: As the number of NSLN+ increased, there was a significant difference in disease-free survival (P = 0.001) and overall survival (P = 0.003) between those patients who had 0 to 4 NSLN+ and those who had 5 or more NSLN+. The SLN+ only patients did not show significant differences with respect to survival, based on the number of SLN+ (overall survival, disease-free survival; P = 0.742). CONCLUSIONS: The survival (overall survival, disease-free survival) for patients with 3 or more SLN+ was not statistically different than for patients with 1 or 2 SLN+ (P = 0.742). However, an alteration of biologic behavior was observed when multiple NSLN+ contain metastatic breast cancer. Involvement of 5 or more NSLN+ portends a significantly (P = 0.001) worse prognosis, regardless of the number of SLN+. PMID- 14553846 TI - The University of Southern California/Van Nuys prognostic index for ductal carcinoma in situ of the breast. AB - BACKGROUND: The original Van Nuys prognostic index (VNPI) was introduced in 1996 as an aid to the complex treatment decision-making process for patients with ductal carcinoma in situ (DCIS) of the breast. This update adds patient age to the previous predictors of local recurrence in breast preservation patients. METHODS: A prospective database consisting of 706 conservatively patients with DCIS was examined using multivariate analysis. Four independent predictors of local recurrence (tumor size, margin width, pathologic classification, and age) were used to derive a new formula for the University of Southern California (USC)/VNPI. RESULTS: In all, 706 patients with pure DCIS were treated with breast preservation. There was no statistical difference in the 12-year local recurrence free survival in patients with USC/VNPI scores of 4, 5, or 6, regardless of whether or not radiation therapy was used (P = not significant). Patients with USC/VNPI scores of 7, 8, or 9 received a statistically significant average 12% to 15% local recurrence-free survival benefit when treated with radiation therapy (P = 0.03). Patients with scores of 10, 11, or 12, although showing the greatest absolute benefit from radiation therapy, experienced local recurrence rates of almost 50% at 5 years. CONCLUSIONS: Ductal carcinoma in situ patients with USC/VNPI scores of 4, 5 or 6 can be considered for treatment with excision only. Patients with intermediate scores (7, 8, or 9) should be considered for treatment with radiation therapy or be reexcised if margin width is less than 10 mm and cosmetically feasible. Patients with USC/VNPI scores of 10, 11, or 12 exhibit extremely high local recurrence rates, regardless of irradiation, and should be considered for mastectomy, generally with immediate reconstruction or reexcision if technically possible. PMID- 14553847 TI - Effect of 0.5 Tesla magnetic resonance imaging on the surgical management of breast cancer patients. AB - BACKGROUND: This trial examines the utility of breast magnetic resonance imaging (MRI) for detection of occult breast disease and its effect on surgical treatment. METHODS: Between October 2000 and March 2002, 76 of 1289 patients underwent bilateral breast MRI within 4 months of a mammogram. The MRI scan, mammogram, pathology reports, and physicians' notes were reviewed to determine impact of MRI on surgical treatment. RESULTS: Magnetic resonance imaging detected 23 additional lesions in 19 patients not detected by mammogram. Cancer occult to mammography was detected by MRI in 6 women, constituting 7.9%. Magnetic resonance imaging impacted surgical treatment plans in 10 of 76 (13.2%) patients. CONCLUSIONS: Magnetic resonance imaging is effective at identifying new subclinical breast disease not seen on mammography. Ten of 76 patients (13.2%) who underwent MRI had their surgical management altered due to MRI findings. Specific criteria should be studied to know which subgroups would benefit most from breast MRI. PMID- 14553848 TI - The effect of neoadjuvant chemotherapy on estrogen and progesterone receptor expression and hormone receptor status in breast cancer. AB - BACKGROUND: Neoadjuvant chemotherapy may decrease tumor volume to allow breast conservation surgery. Its effect on estrogen and progesterone receptor (ER/PR) expression and hormone receptor (HR) status is controversial. METHODS: From February 2001 to July 2002, 56 breast cancer patients treated with neoadjuvant chemotherapy and 56 non-neoadjuvant therapy (control) patients with adequate tissue samples were identified. Quantitative ER/PR expression was analyzed in preneoadjuvant or preoperative core biopsies and final surgical specimens. Changes between the two groups were compared to determine if alterations were due to neoadjuvant chemotherapy or tissue sampling. RESULTS: The ER/PR expression changed in 34 (61%) neoadjuvant chemotherapy patients and 27 (48%) control patients. These expression changes resulted in HR status (positive/negative) alterations in 3 patients (5%) in both groups. Age, histology, chemotherapy regimen, and neoadjuvant response did not predict change. CONCLUSIONS: Hormone receptor status changed in 5% of neoadjuvant chemotherapy and control groups due to tissue sampling. As these changes may impact treatment, HR expression reanalysis in final surgical specimens is recommended. PMID- 14553849 TI - The molecular detection of micrometastatic breast cancer. AB - BACKGROUND: The rapid evolution of molecular technology and novel markers provides the opportunity to establish a more effective means to detect micrometastatic breast cancer. Given the controversies concerning application and clinical relevance, this review critically evaluates the current status of these molecular staging technologies. DATA SOURCES: Breast cancer literature addressing (1). molecular detection methodologies (immunohistochemistry, reverse transcriptase polymerase chain reaction, and microarray analysis); (2). specific tissue applications such as lymph nodes, bone marrow aspirate, and peripheral blood; (3). expert commentary concerning the clinical applications and pitfalls of these technologies; and (4). recent data from our molecular diagnostics laboratory. CONCLUSIONS: Molecular detection technologies such as reverse transcriptase polymerase chain reaction and microarray analyses are being developed that will likely have future application as cancer diagnostics. Further work is needed to establish assays that are validated by prospective clinical studies. Early identification of clinically relevant disease could lead to new treatment or staging approaches for breast cancer. PMID- 14553850 TI - Primary nonphylloides breast sarcomas. AB - BACKGROUND: The prevalence of primary breast sarcoma is low, occurring in fewer than 1% of women with breast malignancies. The purpose of this study was to examine the presentation, treatment, and prognosis of patients presenting with these neoplasms. METHODS: This was a retrospective review of patients with a primary breast sarcoma treated at Mayo Clinic, Rochester, Minnesota, between 1975 and 2001. Follow-up information was obtained. RESULTS: Of the 55 patients, 17 had breast-conserving therapy and 38 women had mastectomy. The mean patient age at presentation was 52 years (range 22 to 82). The types of sarcoma included angiosarcoma (18), malignant fibrous histiocytoma (11), stromal sarcoma (8), liposarcoma (4), leiomyosarcoma (4), dermatofibrosarcoma protuberans (4), osteosarcoma (3), fibrosarcoma (2), and rhabdomyosarcoma (1). Follow-up information was available for 53 patients, with a mean follow-up of 81 months. Twenty-nine of 53 patients (55%) developed recurrent sarcoma, and 23 patients (43%) died of their disease. Twenty-seven patients had no evidence of recurrence, and 3 patients were alive with disease at last follow-up. Overall median survival of patients with breast sarcoma was 58 months. Patients with angiosarcoma had a poorer outcome than other sarcoma patients. Twelve of 18 patients (67%) died of angiosarcoma, compared with 11 of 32 patients (34%) of all other sarcoma patients combined. Of 34 patients who did not receive adjuvant chemotherapy or radiation, 13 died of their disease (38%), as compared with 10 of 16 patients (63%) who did receive adjuvant therapy. CONCLUSIONS: While primary nonphylloides breast sarcomas are rare tumors, their treatment and prognosis are poor. Adjuvant chemotherapy and radiation did not improve survival in this report. Surgical extirpation remains the only effective treatment. PMID- 14553851 TI - Low-risk palpable breast masses removed using a vacuum-assisted hand-held device. AB - BACKGROUND: This study evaluates the safety, efficacy, and patient acceptance of a vacuum-assisted, hand-held biopsy device (Mammatome) in the percutaneous removal of breast masses using ultrasound guidance. METHODS: A multicenter, nonrandomized study evaluated 216 women with low-risk palpable lesions. Lesions 1.5 to 3.0 cm in size were removed using an 8-gauge probe. Those lesions <1.5 cm were removed with the 11-gauge probe. Follow-up evaluation was performed at 10 days and 6 months after biopsy. RESULTS: A total of 127 patients had biopsies using the 8-gauge probe, and 89 patients had biopsies using the 11-gauge probe. At 6-month follow-up, 98% of the lesions remained nonpalpable, 73% with no ultrasonographically visible evidence of the original lesion. Most complications were mild and anticipated. Most patients (98%) were satisfied with incision appearance, and 92% of patients would recommend the procedure to others. CONCLUSIONS: Percutaneous removal of palpable benign breast masses using the Mammotome system is feasible and safe, and yields high patient satisfaction. The results at 6 months after biopsy demonstrated the effectiveness of benign lesion removal, with correlative clinical data demonstrating lack of palpability and no need for additional procedures. Continuing evaluation of long-term efficacy is ongoing. PMID- 14553852 TI - Axillary lymph node metastases in patients with a final diagnosis of ductal carcinoma in situ. AB - BACKGROUND: Recent studies report the incidence of axillary metastases in patients with ductal carcinoma in-situ (DCIS) approaches 13%. The purpose of this study was to define the incidence of axillary micrometastases in patients with pure DCIS before and after the introduction of sentinel lymph node biopsy. METHODS: Patients with a final diagnosis of DCIS form the basis of this study. Data were entered prospectively into an Institutional Review Board approved Oracle database from January 1997 through July 2002. RESULTS: One hundred and thirty-four patients had lymph nodes evaluated. Ninety-eight percent of patients had no evidence of metastatic disease and 2% were found to have micrometastases. This was consistent in those who had level I or II lymph node sampling or both and those who had lymphatic mapping and a sentinel lymph node biopsy procedure. CONCLUSIONS: These data do not support axillary lymph node removal of any type in patients with pure DCIS. PMID- 14553854 TI - Intraoperative ultrasonography guidance is accurate and efficient according to results in 100 breast cancer patients. AB - BACKGROUND: The frequency of nonpalpable breast cancer has doubled in the last 10 years. Surgical use of high quality portable ultrasound units has made it possible to evaluate the time-saving method of intraoperative ultrasonography localization. METHODS: Ultrasonography localization in the operating room immediately prior to definitive surgery was performed by the surgeon. All tissue underwent specimen mammography and ink orientation. The presence of negative margins, the closest margins, the need for reexcision, and the cosmetic result were examined for each patient. RESULTS: Ultrasonography correctly localized all breast cancers at surgery (100% sensitivity). Negative margins for invasive carcinoma were found in 90% of patients. Negative margins were at least 10 mm in 27%, at least 5 mm in 62%, and at least 1 mm in 90%. Positive margins were due to the presence of noncalcified ductal carcinoma in-situ (4 of 10) or multifocal invasive disease (6 of 10) noted at final histology. Reexcisions were performed in 9% of patients. CONCLUSIONS: Surgical use of intraoperative ultrasonography for localization of breast cancer is accurate and efficient. Ultrasonography localization is easier on the patient, is time and cost efficient, and convenient to schedule. Surgeons should embrace this new skill and utilize this tool whenever a nonpalpable breast cancer can be seen with ultrasonography. PMID- 14553853 TI - Intraoperative margin assessment reduces reexcision rates in patients with ductal carcinoma in situ treated with breast-conserving surgery. AB - BACKGROUND: Reported rates of reexcision for margin control after breast conserving surgery for ductal carcinoma in situ (DCIS) range from 48% to 59%. The optimal technique for intraoperative margin assessment in patients with DCIS has yet to be defined. We sought to determine whether intraoperative multidisciplinary evaluation using gross tissue assessment and sectioned-specimen radiography reduces the need for reoperation for margin control in DCIS. METHODS: A prospectively compiled database was used to identify patients who had DCIS diagnosed by core needle biopsy and were treated with breast-conserving surgery at our institution between July 1999 and July 2002. All patients had intraoperative gross margin assessment and specimen radiography of both the whole and sliced specimen for calcifications. RESULTS: Four hundred two patients with DCIS were evaluated at our institution during the study period. Of these, 160 had excisional biopsy for diagnosis prior to referral, 92 had mastectomy as their initial procedure, 40 were seen for a second opinion only, and 1 patient refused surgery. The remaining 109 patients formed the study population. The median age was 55 years (range 34 to 81). The median pathologic size of DCIS was 1.2 cm (range 0.2 to 8.0 cm). Fifty-nine patients had positive (less than 1 mm) or close (less than 5 mm) margins on intraoperative assessment. Final pathology agreed with intraoperative assessment of a positive or close margin in 43 of the 59 patients (P = 0.00005). Seventy-five percent of those thought to have a positive or close margin at the time of surgery (n = 44) underwent intraoperative reexcision. Of the total 109 patients, 31 (34%) had an intraoperative reexcision that resulted in a change in margin status from positive on intraoperative evaluation to negative on final pathologic evaluation (P < 0.00001). A second procedure for margin control was necessary in only 24 patients (22%). The decision to excise additional tissue at the first surgery on the basis of intraoperative assessment resulted in significantly fewer second procedures for margin control (P = 0.029). CONCLUSIONS: In patients with DCIS, intraoperative margin assessment by gross pathological examination and sliced specimen radiography significantly affects intraoperative decision making, and excision of further tissue on the basis of intraoperative assessment results in a substantial decrease in second procedures for margin control. PMID- 14553855 TI - Postoperative ultrasound-guided percutaneous placement of a new breast brachytherapy balloon catheter. AB - BACKGROUND: The optimal technique and timing for the implantation of a recently developed breast brachytherapy balloon catheter (MammoSite; Proxima Therapeutics, Alpharetta, Georgia) have not been well defined. We hypothesized that placing this postoperatively, utilizing percutaneous ultrasound-guided placement, would be preferable. METHODS: Patients who met eligibility requirements for breast brachytherapy were implanted with the MammoSite device utilizing percutaneous ultrasound-guided technique. Additionally, to study optimal timing, a historical cohort of patients operated upon for breast cancer by two of the authors were analyzed to compare the intraoperative and postoperative candidacy for MammoSite placement. RESULTS: Twenty-one of 23 patients successfully completed brachytherapy after implantation, with only 2 (9%) requiring catheter removal secondary to unfavorable balloon positioning. There were no serious complications. Of 343 historical patients with breast cancer, 137 were intraoperative candidates for brachytherapy, but final postoperative histology disqualified 40 (29%). CONCLUSIONS: Implantation of the MammoSite brachytherapy device is optimally performed postoperatively, after the final pathology is defined, utilizing ultrasound-guided percutaneous technique. PMID- 14553856 TI - Single-step ultrasound localization of breast lesions and lumpectomy procedure. AB - BACKGROUND: This review was made to evaluate whether the efficiencies of intraoperative ultrasound-guided localization could be extended to stereotactic biopsy cases by using a marker visible by sonography and mammography. METHODS: A retrospective review identified 170 stereotactic directional vacuum-assisted biopsy (DVAB) procedures marked with an ultrasound-visible marker. Localization device, imaging method, lesion retrieval, and margin status were assessed for patients having subsequent lumpectomy or wider excision. RESULTS: Nineteen of 170 patients underwent lumpectomies (12) or wider excision (7) localized by a radiofrequency device or 18G needle up to 7 weeks after stereotactic biopsy. In 15 of 19 procedures, an ultrasound-guided localization was performed intraoperatively, targeting the marker. All targeted lesions were successfully excised. In the 13 malignant lesions (of 19 surgeries), only 1 had a positive margin (8%). CONCLUSIONS: Marking stereotactic biopsies with a sonographically visible marker allows ultrasound-guided intraoperative localization, improving efficiencies for the patient, surgeon, and operating room schedule. PMID- 14553857 TI - The effect of ipsilateral whole breast ultrasonography on the surgical management of breast carcinoma. AB - BACKGROUND: Patients can be selected for breast conservation therapy using mammography and physical examination. Whole breast ultrasonography has been shown to identify lesions not seen on mammography. This study sought to determine how often whole breast ultrasonography changes the surgical management of breast cancer. METHODS: All patients with stage 1 and 2 breast cancer undergoing whole breast ultrasonography were identified. A change in surgical management was defined as the identification by ultrasonography alone of foci of carcinoma greater than 1 cm from the primary tumor site or in another quadrant of the breast. RESULTS: There were 1385 breast cancer patients; 31% had ultrasonography. Eighteen percent of patients had abnormalities identified by ultrasonography alone. Changes in management occurred in 2.8%. The additional lesions led to four wider resections and eight mastectomies. Patients with ultrasonography abnormalities were significantly younger, and more likely to have histologic grade 2 or 3 disease. CONCLUSIONS: These findings do not support the routine use of ultrasonography in all breast cancer patients. Significant abnormalities were more commonly seen among younger patients with higher grade lesions. PMID- 14553858 TI - Does ultrasound core breast biopsy predict histologic finding on excisional biopsy? AB - BACKGROUND: The purpose of this study was to determine whether ultrasound-guided core breast biopsy accurately predicts the histologic finding of a subsequent excisional procedure. METHODS: Data were collected prospectively from 1997 to 2001 for 832 ultrasound-guided core breast biopsies (USB) that were followed by excisional breast procedure (EP) within 1 year at our institution. The principal histologic finding obtained at USB and EP was identified for each procedure and the degree of agreement was assessed. RESULTS: The USB histology predicted EP histology in 90% (n = 746) of the procedures. The USB histology was more significant than EP histology in 3% (n = 22) of procedures; USB histology underdetermined EP histology in 7% (n = 64) of procedures. Overall, our results indicate moderate agreement between the principal histology identified at USB relative to that identified at EP. CONCLUSIONS: Ultrasound-guided core breast biopsy is an effective diagnostic method, but sampling limitations do exist. PMID- 14553859 TI - Indigent breast cancer patients among all racial and ethnic groups present with more advanced disease compared with nationally reported data. AB - BACKGROUND: This study examines the epidemiologic and pathologic characteristics of indigent breast cancer patients followed up in a public city hospital in comparison to national standards. METHODS: A prospective oncology database was queried to identify all patients presenting with primary breast cancer. Medical records of 188 patients identified between March 1997 and May 2002 were retrospectively reviewed. Pathologic and epidemiologic data were compared with 1998 data reported by the Surveillance, Epidemiology, and End Results (SEER) program. RESULTS: Among the patient population 10% were Caucasian, 13% African American, 49% Hispanic, 25% Chinese, and 6% were of other background. The majority of patients were uninsured. Indigent patients within each ethnic group presented with more advanced disease when compared with patients reported by SEER. CONCLUSIONS: Indigent patients among all ethnic and racial backgrounds present with more advanced disease when compared with national statistics reported by SEER. The majority of these patients is uninsured and would benefit from more aggressive education, screening, detection methods, and follow-up. PMID- 14553860 TI - Breast cancer severity score is an innovative system for prognosis. AB - BACKGROUND: Survival in breast cancer maybe more accurately predicted by a scoring system based on the biologic characteristics of the tumor such as diameter, number of involved nodes, and hormone receptor status than by conventional staging. METHODS: Medical records of 419 patients with a diagnosis of breast cancer from a university hospital between 1997 and 2002 were reviewed. Tumor characteristics were given a value reflecting the impact on survival. Tumor diameter was assigned 1 point per cm, specifically: 1 point for 0.1 to 1 cm, 2 points for 1.1 to 2 cm, 3 points for 2.1 to 5 cm, and 4 points for 5.1 cm and over. Nodal status was assigned 1 point if the patient had 1 to 3 positive nodes, 5 points if 4 to 10 positive nodes were found, and 10 points if more than 10 nodes were identified. Hormone receptor status was given 1 point if either estrogen or progesterone receptors were not present or 2 points if both were absent. The breast cancer severity score (BCSS) was used to analyze overall and disease-free survival by Kaplan-Meier analysis. Significance of differences in survival was determined by log-rank analysis. RESULTS: An increased severity score was inversely proportional to overall and disease-free survivals. A BCSS of greater than 7 was most predictive of outcome with 5-year survival of 68%; and a BCSS of less than 7 had a 5-year survival of 98% (P = 0.0028). A BCSS of greater than 7 also discriminated disease-free survival within stage II (P = 0.02) and stage III (P = 0.01). CONCLUSIONS: The BCSS provides a better resolution of outcome than traditional staging, and works both for overall survival and within specific stages. Using the BCSS, management and follow-up can be specifically tailored for low- and high-risk patients. PMID- 14553861 TI - An increased risk of breast cancer after delayed first parity. AB - BACKGROUND: As delayed childbirth increases for socioeconomic and fertility reasons, its impact on breast cancer risk needs definition. METHODS: From 1975 to 1981, 1307 women with childbirth at >or=40 years of age were identified. They were divided into four groups by estimated first birth median ages (EFBMA): 23, 34, 38, and 41 years, corresponding to previous parity of more than 3, 2 or 3, 1, and zero, respectively. Cancer Registry cross-referencing identified those diagnosed with breast cancer. RESULTS: Breast cancer developed in 39 women. The EFBMA of 41 years carried a relative risk of 3.7, (95%CI: 1.30 to 10.5) compared with age 23. Odds ratio of breast cancer was 1.08 (95%CI: 1.02 to 1.14) with each year older at first birth and 0.79 (95% CI: 0.67 to 0.93) for each additional previous birth. CONCLUSIONS: Increased breast cancer risk with advancing maternal age at first childbirth is supported by 3.7 relative risk in women with an EFBMA of 41 years compared with those with an EFBMA of 23 years. PMID- 14553862 TI - Encircling guidewire facilitates complete excision of image-localized breast lesions. AB - BACKGROUND: An innovative encircling guidewire defines three sides of a target lesion with a single puncture to achieve negative margins. METHODS: Twenty-five patients requiring image-guided surgery were localized with an encircling guidewire. Although the deployed wire is circular, it is straight when placed, using a straight outer needle. After image-guided placement of the wire around the lesion, all patients underwent standard surgical excision. Each patient was categorized by proper localization of the target lesion, presence of negative margins, closest margin, and need for reexcision. RESULTS: The circlewire wire identified the target lesion in all cases. There were no complications relative to either version of the leading tip on the guidewire. Negative margins were achieved in all pure invasive ductal carcinomas. Positive margins were found in all 3 patients with extensive noncalcified in-situ disease and 1 patient with multifocal invasive lobular carcinoma. CONCLUSIONS: An innovative encircling localizing guidewire device gives the surgeon a new option to completely remove a nonpalpable breast lesion. PMID- 14553864 TI - Type 2 diabetes, cardiovascular risk, and the link to insulin resistance. AB - BACKGROUND: Patients with type 2 diabetes mellitus frequently have coexistent dyslipidemia, hypertension, and obesity, and are at risk for microvascular and macrovascular disease complications such as myocardial infarction, stroke, retinopathy, and microalbuminuria. To optimize cardiovascular health outcomes for patients with type 2 diabetes, strategies to reduce the risks of microvascular and macrovascular disease are needed in clinical practice. OBJECTIVE: This article provides an overview of the cardiovascular risk profile of patients with type 2 diabetes and discusses the cardiovascular consequences of use of the thiazolidinediones (insulin-sensitizing agents) in the treatment of type 2 diabetes. METHODS: A literature search of MEDLINE/PubMed was performed to identify relevant articles published from 1966 to April 2003. Search terms used were diabetes, cardiovascular disease, atherosclerosis, dyslipidemia, obesity, hypertension, blood pressure, hyperglycemia, inflammation, C-reactive protein, fibrinolysis, plasminogen activator inhibitor type-1, microalbuminuria, thiazolidinediones, safety, hepatotoxicity, and edema. Bibliographies within the identified articles were also evaluated for additional relevant articles and information. RESULTS: Recommendations for cardiovascular risk reduction through preventive and therapeutic strategies that target the symptoms of insulin resistance may reduce the microvascular and macrovascular sequelae of diabetes and ameliorate the impact of other components of the metabolic syndrome, including hypertension, hyperglycemia, and obesity. In this regard, thiazolidinediones are promising therapies. CONCLUSIONS: Early data suggest that, in addition to reducing hyperglycemia, pioglitazone and rosiglitazone effect changes in the dyslipidemic profile, hemodynamics, vascular inflammation, and endothelial functioning of patients with type 2 diabetes. Additional research is needed to further distinguish the cardiovascular benefits of these drugs. PMID- 14553865 TI - Pancreatic beta-cell loss and preservation in type 2 diabetes. AB - BACKGROUND: In most individuals, the need to respond to progressive states of insulin resistance is met by increasing insulin production. For insulin-resistant patients, however, the balance between insulin supply and demand may fail from the progressive loss of pancreatic beta-cell function, eventually leading to type 2 diabetes mellitus. OBJECTIVE: The aim of this review was to discuss the current concepts underlying potential pancreatic beta-cell failure in the progression toward type 2 diabetes and therapies that may alter the process. METHODS: Data included in this review were identified through a MEDLINE search for articles published from 1966 to April 2003. Search terms used were beta cell, diabetes, insulin resistance, obesity, cardiovascular disease, thiazolidinediones, and metformin. RESULTS: Evidence of the progressive loss of beta-cell function may include altered conversion of proinsulin to insulin, changes in pulsed and oscillatory insulin secretion, and quantitative reductions in insulin release. Potential underlying mechanisms are glucose toxicity, lipotoxicity, poor tolerance of increased secretory demand, and a reduction in beta-cell mass. CONCLUSION: Current clinical management of type 2 diabetes is focused on treatment of the signs and symptoms of late-stage disease rather than addressing potential underlying causes, which may be amenable to currently available therapies, based on a broad understanding of existing data, practice experience, and rational speculation. PMID- 14553866 TI - Insulin resistance: from predisposing factor to therapeutic target in type 2 diabetes. AB - BACKGROUND: Insulin resistance contributes to the pathogenesis of type 2 diabetes and is closely linked with cardiovascular risk factors and premature cardiovascular disease. OBJECTIVE: The purpose of this paper was to review the importance of insulin resistance as a core defect in type 2 diabetes, a potential contributor to accelerated atherosclerosis, and a potential target for insulin sensitizing agents. METHODS: Articles considered for inclusion in this review were identified through a search of MEDLINE/PubMed for reports published from 1966 to April 2003. Search terms used were insulin resistance, diabetes, insulin sensitivity, obesity, cardiovascular disease, metformin, thiazolidinediones, pioglitazone, rosiglitazone, and troglitazone. RESULTS: An overview of the epidemiology, natural history, and pathophysiology of type 2 diabetes is provided, with a focus on insulin resistance and a related discussion of the impact of current therapies used to treat insulin-resistant patients. In particular, information on insulin-sensitizing agents-metformin and the currently available thiazolidinediones (TZDs), pioglitazone and rosiglitazone-is presented. Although metformin has been shown to indirectly reduce insulin resistance, TZDs are the only available agents that have been shown to directly lower insulin resistance. CONCLUSIONS: Recent evidence indicates that metformin, pioglitazone, and rosiglitazone may improve the dyslipidemic profile, reduce vascular inflammation, and improve endothelial dysfunction, all of which may be particularly important to physicians seeking treatment options to prevent or reduce cardiovascular complications in patients with type 2 diabetes. PMID- 14553867 TI - Multicenter retrospective assessment of thiazolidinedione monotherapy and combination therapy in patients with type 2 diabetes: comparative subgroup analyses of glycemic control and blood lipid levels. AB - BACKGROUND: Thiazolidinediones (TZDs) have contributed to the management of patients with type 2 diabetes mellitus as unique insulin-sensitizing agents. When used as monotherapy or in combination therapy, these drugs not only reduce glycosylated hemoglobin (HbA(1c)) levels, but also effect changes in blood lipid concentrations and have the potential to ameliorate cardiovascular disease risk. Although drugs in the TZD class are perceived to be equivalent clinically, prospective and retrospective studies have demonstrated their ability to modify blood lipid levels. OBJECTIVE: We evaluated and compared the effects of pioglitazone and rosiglitazone monotherapy and combination therapy on blood lipid levels and HbA(1c) in patients with type 2 diabetes. METHODS: We conducted a multicenter retrospective chart review of 1115 records of patients with type 2 diabetes who received pioglitazone or rosiglitazone, alone or in combination with other antidiabetic agents, between August 1, 1999, and August 31, 2000. The review was conducted to evaluate pretreatment and posttreatment levels of triglyceride, total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and HbA(1c). RESULTS: All observed demographic characteristics, comorbidities, and concomitant drug use were similar in both treatment cohorts. Of the patients who received pioglitazone, 83% also received >/=1 other antihyperglycemic agent and 59% received some form of antihyperlipidemic therapy. Among those who received rosiglitazone, 81% received concomitant antihyperglycemic medication and 60% received some form of antihyperlipidemic therapy. With pioglitazone, mean levels of serum triglyceride, total cholesterol, and LDL-C decreased and HDL-C increased in most patients, with or without concomitant antihyperglycemic medications; with rosiglitazone, with or without other antidiabetic agents, triglyceride and HDL-C levels decreased, whereas total cholesterol and LDL-C levels increased in most patients. Reductions in HbA(1c) levels and increases in body weight related to each study drug were comparable. CONCLUSIONS: This comparative assessment of pioglitazone and rosiglitazone, based on observational data, reveals that use of these TZDs with other antidiabetic agents was similar in 605 primary care practices in the United States. In both monotherapy and combination treatment regimens, pioglitazone was associated with greater beneficial effects on lipids than was rosiglitazone. Additional studies are needed to determine the long-term outcomes of TZD therapy with concomitant antihyperglycemic medications. PMID- 14553868 TI - Implementation of an evidence-based guideline to reduce duration of intravenous antibiotic therapy and length of stay for patients hospitalized with community acquired pneumonia: a randomized controlled trial. AB - PURPOSE: Patients with pneumonia often remain hospitalized after they are stable clinically, and the duration of intravenous antibiotic therapy is a rate-limiting step for discharge. The purpose of this study was to determine whether implementation of an evidence-based guideline would reduce the duration of intravenous antibiotic therapy and length of stay for patients hospitalized with pneumonia. METHODS: In a seven-site, cluster randomized clinical trial, we enrolled 325 control and 283 intervention patients who were admitted by one of 116 physician groups. Within site, physician groups were assigned randomly to receive a practice guideline alone (control arm) or a practice guideline that was implemented using a multifaceted strategy (intervention arm). The effectiveness of guideline implementation was measured by the duration of intravenous antibiotic therapy and length of stay; differences in the rates of discontinuation and hospital discharge were assessed with proportional hazards models. Medical outcomes were assessed at 30 days. RESULTS: Intravenous antibiotic therapy was discontinued somewhat more quickly in the intervention group (hazard ratio [HR] =1.23; 95% confidence interval [CI]: 1.00 to 1.52; P = 0.06) than in the control group. Intervention patients were discharged more quickly, but the difference was not statistically significant (HR = 1.16; 95% CI: 0.97 to 1.38; P = 0.11). Fewer intervention (55% [157/283]) than control (63% [206/325]) patients had medical complications during the index hospitalization (P = 0.04), with no differences in other medical outcomes, including mortality, rehospitalization, and return to usual activities, between treatment arms. CONCLUSIONS: The multifaceted guideline implementation strategy resulted in a slight reduction in the duration of intravenous antibiotic therapy and a nonsignificant reduction in length of stay, without affecting patient outcomes. PMID- 14553869 TI - Antibiotic-coated hemodialysis catheters for the prevention of vascular catheter related infections: a prospective, randomized study. AB - PURPOSE: To determine the efficacy of minocycline-rifampin-coated hemodialysis catheters in reducing catheter-related infections in patients requiring hemodialysis for acute renal failure. METHODS: Between May 2000 and March 2002, 66 patients were randomly assigned to receive a minocycline-rifampin-impregnated central venous catheter and 64 were randomly assigned to receive an unimpregnated catheter. Patients were followed prospectively until the catheter was removed. Catheter-related infection was determined through quantitative catheter cultures, quantitative blood cultures, or both. RESULTS: Both groups of patients were similar in age, sex, underlying disease, type of dialysis (continuous vs. intermittent), neutropenia during catheterization and its duration, catheter insertion difficulties, and administration of blood products or medication. The mean (+/- SD) catheter dwell time was the same in both groups (8 +/- 6 days, P = 0.7). There were seven catheter-related infections (11%), all associated with the use of unimpregnated catheters. Kaplan-Meier estimates for the risk of catheter related infection showed that coated catheters were less likely to be associated with infection (P = 0.006). CONCLUSION: The use of polyurethane hemodialysis catheters impregnated with minocycline and rifampin decreases the risk of catheter-related infection in patients with acute renal failure. PMID- 14553870 TI - Conventional and molecular epidemiology of trimethoprim-sulfamethoxazole resistance among urinary Escherichia coli isolates. AB - BACKGROUND: Antibiotic resistance is increasing in Escherichia coli, the most common cause of urinary tract infections, but its epidemiology has not been well described. We evaluated the epidemiology of trimethoprim-sulfamethoxazole resistant E. coli in a large, public health care system in Denver, Colorado. METHODS: Outpatients with E. coli urinary tract infections during the first 6 months of 1998 were evaluated retrospectively. A prospective study was then performed to confirm the rate of trimethoprim-sulfamethoxazole resistance. We used several strain-typing methods (pulsed-field gel electrophoresis, ribotyping, serotyping) to evaluate the molecular epidemiology of the resistance. RESULTS: The rate of trimethoprim-sulfamethoxazole resistance was similar in the retrospective (24% [161/681]) and prospective (23% [30/130]) phases of the study (P = 0.89). Almost all trimethoprim-sulfamethoxazole-resistant strains (98%) were resistant to at least one other antibiotic. Risk factors for infection with a resistant strain included age < or =3 years, Hispanic ethnicity, recent travel outside the United States, and a prior urinary tract infection. However, rates of resistance were >15% among nearly all of the subgroups. Most strains had high level resistance (>1000 microg/mL) to trimethoprim-sulfamethoxazole. Of the 23 resistant isolates evaluated, 10 (43%) belonged to the clone A group. There was no correlation between conventional epidemiologic characteristics and the molecular mechanism of resistance or strain type. CONCLUSION: Resistance to trimethoprim-sulfamethoxazole among E. coli isolates among patients in a Denver public health care system is common, with high rates of resistance even among patients without risk factors. PMID- 14553871 TI - Interferon gamma levels in pleural fluid for the diagnosis of tuberculosis. AB - PURPOSE: To assess the utility of interferon gamma levels, including identification of the best cutoff for the diagnosis of tuberculosis. METHODS: We prospectively studied consecutive patients in a tertiary care, university affiliated hospital who had pleural effusions. Interferon gamma levels were measured blindly by radioimmunoassay. The diagnosis of tuberculosis was established using prespecified standard criteria. RESULTS: Of the 595 patients with pleural effusions, 82 patients (14%) had tuberculosis. The area under the receiver operating characteristic (ROC) curve for elevated interferon gamma levels in the diagnosis of tuberculosis was 0.99 (95% confidence interval [CI]: 0.97 to 1.00). A cutoff of 3.7 IU/mL yielded a sensitivity of 0.98 (95% CI: 0.91 to 1.00) and a specificity of 0.98 (95% CI: 0.96 to 0.99). The areas under the ROC curves, and the test's sensitivity and specificity, were similar among patients of different ages and by percentage of lymphocytes in the pleural fluid. In 5 of the 28 patients with hematologic malignancies, interferon gamma levels were slightly above the cutoff; no patient with vasculitis or granulomatous diseases had levels higher than 3.7 IU/mL. The 14 immunocompromised patients and the 3 transplantation patients with tuberculosis had interferon gamma levels greater than the cutoff. CONCLUSION: Elevated pleural interferon gamma levels (>3.7 IU/mL) are very valuable in diagnosing pleural tuberculosis. Patients with pleural effusion due to hematologic neoplasms occasionally have levels slightly above the cutoff. PMID- 14553872 TI - Reduction in long-term functional disability in rheumatoid arthritis from 1977 to 1998:a longitudinal study of 3035 patients. AB - PURPOSE: If newer, more aggressive treatment strategies in rheumatoid arthritis are more effective, long-term outcomes in rheumatoid arthritis should be improving substantially. We therefore assessed trends in disability over time in a large cohort of patients with rheumatoid arthritis. METHODS: We examined functional disability data from 3035 patients with rheumatoid arthritis whose disease onset was from 1977 to 1998. Disability data were collected semiannually with the Health Assessment Questionnaire disability index. We then estimated average disability for each patient. We also computed mean disability for each calendar year by averaging the values from all patients in that year. We examined the relation of successive annual cohorts and subsequent disability, adjusting for age, sex, race, education, clinical center, disease duration, follow-up, and attrition. We used two regression approaches: ordinary and generalized least squares. RESULTS: Average disability declined by about 2% to 3% per calendar year of disease onset (2.7% to 2.8% per year [P <0.001] in univariable models and 2.0% to 2.1% per year [P <0.001] in multivariable models). This trend was consistent by age, sex, race, disease duration, clinical center, and baseline disability. CONCLUSION: After accounting for potential confounders, average disability levels in rheumatoid arthritis have declined by approximately 40% in the 20+ years since 1977. This decline is consistent with a beneficial effect of the associated changes in treatment strategies. PMID- 14553873 TI - Inhaled corticosteroid use in asthma and the prevention of myocardial infarction. AB - BACKGROUND: Asthma patients may be at increased risk of cardiovascular outcomes due to hypoxemia from asthma exacerbations and bronchodilator-induced tachycardia. We investigated whether inhaled corticosteroids, which are known to improve asthma control and reduce exacerbations, are associated with a lower rate of myocardial infarction. METHODS: We used the Saskatchewan Health databases to form a population-based cohort of subjects aged 5 to 44 years who were using antiasthma drugs between 1975 and 1991. Subjects were followed until 1997, the age of 55 years, or death. A nested case-control approach was used where each subject with a first myocardial infarction was matched on calendar time, age, and sex with up to 10 controls randomly selected from the cohort. RESULTS: The cohort consisted of 30,569 subjects, including 105 patients with myocardial infarction who were matched with 933 controls. The adjusted rate ratio of myocardial infarction for inhaled corticosteroid use during the year before the index date was 0.56 (95% confidence interval [CI]: 0.32 to 0.99) as compared with no use. Myocardial infarction decreased by 12% (95% CI: 0% to 23%) with each additional canister used during this 1-year period. The rate ratio of myocardial infarction for inhaled corticosteroid use was 0.78 (95% CI: 0.41 to 1.51) among patients with milder asthma and 0.19 (95% CI: 0.04 to 0.97) among those with more severe asthma. CONCLUSION: Inhaled corticosteroid use may reduce the risk of myocardial infarction in asthma patients, particularly those with more severe disease. PMID- 14553874 TI - Cost-effectiveness of bypass surgery versus stenting in patients with multivessel coronary artery disease. AB - PURPOSE: To compare the cost-effectiveness of surgical and angioplasty-based coronary artery revascularization techniques, in particular, angioplasty with primary stenting. METHODS: We used data from the Study of Economics and Quality of Life, a substudy of the Bypass Angioplasty Revascularization Investigation (BARI), to measure the outcomes and costs of angioplasty and bypass surgery in patients with multivessel coronary artery disease who had not undergone prior coronary artery revascularization. Using a Markov decision model, we updated the outcomes and costs to reflect technology changes since the time of enrollment in BARI, and projected the lifetime costs and quality-adjusted life-years (QALYs) for the two procedures from the time of initial treatment through death. We accounted for the effects of improved procedural safety and efficiency, and prolonged therapeutic effects of both surgery and stenting. This study was conducted from a societal perspective. RESULTS: Surgical revascularization was less costly and resulted in better outcomes than catheter-based intervention including stenting. It remained the preferred strategy after adjusting the stent outcomes to eliminate the costs and events associated with target lesion restenosis. Among angioplasty-based strategies, primary stent use cost an additional 189,000 US dollars per QALY gained compared with a strategy that reserved stent use for treatment of suboptimal balloon angioplasty results. CONCLUSION: Bypass surgery results in better outcomes than angioplasty in patients with multivessel disease, and at a lower cost. PMID- 14553875 TI - Autoimmune aspects of cytokine and anticytokine therapies. AB - Cytokines and anticytokines are used increasingly in the treatment of immune, autoimmune, inflammatory, infectious, and malignant disorders. Commonly used treatments include the anti-tumor necrosis factor agents interferon alpha, interferon beta, interferon gamma, and interleukin 2. Several autoimmune phenomena have been reported in patients treated with these substances. This review summarizes the published data on the autoimmune manifestations associated with cytokine and anticytokine therapies, as well as describes possible mechanisms of these phenomena. PMID- 14553876 TI - Underuse of osteoporosis medications in elderly patients with fractures. PMID- 14553877 TI - Association between mortality and occupation among movie directors and actors. PMID- 14553878 TI - Cases from the Osler Medical Service at Johns Hopkins University. PMID- 14553879 TI - Making evidence-based practice improvement more than a comforting sentiment. PMID- 14553880 TI - Coronary artery bypass surgery is still cost-effective: using calibrated models to update clinical trials. PMID- 14553881 TI - Functional class in patients with heart failure is associated with the development of diabetes. PMID- 14553883 TI - Is chronic autoimmune thyroiditis a systemic disease? PMID- 14553885 TI - Antiglomerular basement membrane autoantibodies are nonpathogenic in Wegener's granulomatosis. PMID- 14553886 TI - Effective prevention of 5-fluorouracil-induced superficial phlebitis by ketoprofen lysine salt gel. PMID- 14553887 TI - Successive thyroid storms treated with L-carnitine and low doses of methimazole. PMID- 14553888 TI - Sjogren's-like syndrome following intravesical bacillus Calmette-Guerin immunotherapy. PMID- 14553890 TI - Rhabdomyolysis associated with probable SARS. PMID- 14553889 TI - Clinical and biological distinctions between type I and type II acquired angioedema. PMID- 14553891 TI - The internal medicine clerkship in the clinical education of medical students. PMID- 14553892 TI - Specific differentiation of recombinant PVY(N:O) and PVY(NTN) isolates by multiplex RT-PCR. AB - The recombinant isolates of tobacco veinal necrotic strain of Potato virus Y (PVYN) and potato tuber necrotic group (PVY(NTN)) contain segments of the PVYO and the PVY(N) genome. Three major recombinant junctions (RJ) are present in the genome of the recombinant PVY(NTN) at sites HC/Pro-P3, 6K2-NIa, and the C terminal region of CP gene and one RJ at HC/Pro-P3 site in some recombinant PVYN isolates (termed PVY(N:O)). Protocols for specific differentiation of the recombinant PVY(NTN) and PVY(N:O) from the non-recombinant PVYN are described. Specific primer pairs were designed to target the three RJs so that sense and antisense primers completely matched the nucleotide sequences at either side of the RJ. In a uniplex reverse transcription-polymerase chain reaction (RT-PCR), the first primer pair amplified a fragment of 641bp from the recombinant PVY(NTN) and PVY(N:O). The second and third primer pairs exclusively amplified fragments of 448 and 290bp, respectively from the recombinant PVY(NTN). In a multiplex (triplex) RT-PCR, when all three primer pairs were used simultaneously, the three fragments (641, 448 and 290bp) were amplified exclusively from the recombinant PVY(NTN), while only one fragment (641bp) was amplified from the PVY(N:O) isolates, clearly differentiating the two recombinant isolates. No amplification was observed from the non-recombinant PVY, including PVYO and North American (NA) PVY(N/NTN). For further improvement of the multiplex RT-PCR, effects of cDNA preparation using specific antisense primers, random primers or oligo(dT) plus random primers were investigated. The cDNA prepared by random primer plus oligo(dT) increased the overall band intensity. PMID- 14553893 TI - Reconciling HIV incidence results from two assays employed in the serological testing algorithm for recent HIV seroconversion (STARHS). AB - The Abbott HIVAB and Vironostika HIV-1 Microelisa assays have both been validated for use in the serological testing algorithm for recent HIV seroconversion (STARHS). This ability to identify recently-acquired infection provides valuable insight into the epidemic. The availability of each assay during different periods led to longitudinal studies of annual HIV incidence being based on a mixture of results from each. We investigated whether results from both assays could be reconciled. Using statistical methods, the correlation of the two assays' results and other performance characteristics were examined. Of 378 anti HIV-1 positive specimens examined by both assays, the Abbott assay flagged 40 as from recent infections, whereas Vironostika flagged 50. The correlation coefficient between screening reactivities in each assay was 0.84, and 0.77 in confirmatory mode. Abbott screening results were significantly higher than its confirmatory results, and some specimens from recent infections may consequently have gone undetected by that assay. This problem was not found with the Vironostika assay. Observational data indicated that the estimated HIV incidence derived from HIVAB results increased as the assay threshold, with its pre-defined seroconversion window, was increased. For Vironostika, the estimated HIV incidence remained stable over a wide range of thresholds. Modelling of the observed relationship between the two assays allowed an estimate of the equivalent threshold in the alternative assay, thus providing a means of reconciling results. Our findings suggest that the Vironostika assay is more reliable than the HIVAB, is easier to use, and is able to allow processing of more specimens per run. PMID- 14553894 TI - Detection of melon necrotic spot virus in water samples and melon plants by molecular methods. AB - Melon necrotic spot virus (MNSV) is a water and soil-borne pathogen affecting species of the Cucurbitaceae family both in hydroponic and soil crops. Molecular methods for detecting MNSV in water samples, nutrient solutions and melon plants were developed. For this purpose, water samples from a water source pool of a hydroponic culture or from the recirculating nutrient solution were concentrated by ultracentrifugation or PEG precipitation followed by RT-PCR analysis. Both concentration methods were suitable to allow the detection of MNSV and represent, as far as we know, the first time that this virus has been detected in water samples. A non-isotopic riboprobe specific for MNSV was obtained and used to detect the virus in plant tissue. Different parts of mechanically infected plants were examined including the roots, stems, inoculated cotyledons and young leaves. Excluding the inoculated cotyledons, the tissues showing the highest accumulation levels of the virus were the roots. The potential inclusion of such tools in management programs is discussed. PMID- 14553895 TI - An efficient method for precise gene substitution in the AcMNPV genome by homologous recombination in E. coli. AB - The RecA-mediated homologous recombination method was improved and used to direct gene replacement in baculoviruses. With this method, the p74 gene in the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) genome was substituted precisely by the p74 gene of Spodoptera litura multicapsid nucleopolyhedrovirus (SpltMNPV). In the recombinant bacmid, the AcMNPV p74 gene promoter controlled directly the expression of SpltMNPV p74 gene. Results of RT PCR showed transcription of SpltMNPV p74 gene in the recombinant, implying the potential use of this easy and efficient method in baculovirus gene function research. PMID- 14553896 TI - IgG-sandwich and IgM-capture enzyme-linked immunosorbent assay for the detection of antibody to Rift Valley fever virus in domestic ruminants. AB - The recent occurrence of the first confirmed outbreaks of Rift Valley fever in humans and livestock outside the African region, namely in the Kingdom of Saudi Arabia and Yemen, is of global medical and veterinary concern. Disadvantages of classical techniques for serological diagnosis of Rift Valley fever include health risk to laboratory personnel, restrictions for their use outside endemic areas and inability to distinguish between different classes of immunoglobulins. We report on the development and validation of sandwich and capture ELISAs (both based on inactivated antigen) for detection of IgG and IgM antibody to Rift Valley fever virus in bovine, caprine and ovine sera. Compared to virus neutralisation and haemagglutination-inhibition tests, the IgG sandwich ELISA was more sensitive in detection of the earliest immunological responses to infection or vaccination with Rift Valley fever virus. Its sensitivity and specificity derived from field data sets ranged in different ruminant species from 99.05 to 100% and from 99.1 to 99.9%, respectively. The specificity of IgM-capture ELISA varied between different species from 97.4 to 99.4%; its sensitivity was 100% in sheep tested 5-42 days post-infection. Our results in field-collected, experimental and post-vaccination sera demonstrate that these assays will be useful for epidemiological surveillance and control programmes, import/export veterinary certification, early diagnosis of infection, and for monitoring of immune response in vaccinated animals. As highly accurate and safe tests, they have the potential to replace traditional diagnostic methods, which pose biohazard risks limiting their use outside of endemic areas to high containment facilities. PMID- 14553897 TI - A real-time PCR assay for the detection of varicella-zoster virus DNA and differentiation of vaccine, wild-type and control strains. AB - Varicella-zoster vaccine is a live attenuated virus. It is, therefore, necessary to have a test to differentiate vaccine from wild-type varicella-zoster virus (VZV) strains for the investigation of varicella or zoster-like rash illness in individuals vaccinated previously. In addition, it is necessary to have a rapid VZV assay for use in the context of smallpox bioterrorism laboratory testing. Using specific primers and hybridization probes, a rapid method to differentiate vaccine strain VZV from wild-type VZV was developed based on the presence or absence of a Pst I restriction site within open reading frame (ORF) 38. Using this ORF 38 assay in conjunction with a similar previously described ORF 62 assay allows for further differentiation of vaccine strain, wild-type and a laboratory control strain (Ellen) VZV. This is accomplished because Ellen VZV is similar to wild-type VZV with respect to the ORF 38 assay but is similar to vaccine strain VZV with respect to the ORF 62 assay. The hybridization probes for each ORF are labeled with different fluorescent tags thus allowing both assays to be run simultaneously in a single tube. Both assays demonstrate a high degree of specificity for VZV and can reliably detect between 10 and 100 copies of VZV DNA. Thus, the real-time polymerase chain reaction (PCR) assay for VZV described below provides a rapid assay allowing the simultaneous differentiation of vaccine, wild type and laboratory control strains of VZV. PMID- 14553898 TI - The opioid receptors in inner ear of different stages of postnatal rats. AB - There is increasing evidence that the opioid system has a role in hearing. To provide further evidence for such a role, the expression of opioid receptor mRNAs and proteins in the inner ear of rats was studied during development from birth (P0) to postnatal day 16 (P16). A semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was employed to detect changes in the expression of delta- (DOR) kappa- (KOR) and mu- (MOR) opioid receptor mRNAs in rat cochleae at P0, P4, P8 and P16. Expression of DOR mRNA levels steadily increased from P0 to P8 with no further increases by P16. KOR mRNA was expressed at a relatively high level at P0 and P4 followed by a decrease while MOR mRNA was expressed at a low level at P0 and P4 followed by an increase by P8 and P16. Immunocytochemical labelling of inner ear sections revealed unique developmental and distribution patterns of opioid receptors. In the organ of Corti DOR immunoreactivity (DOR-IR) was detected in hair cells from P4. In contrast MOR-IR was present only in supporting cells at P0-P16. In the spiral ganglion all three receptor subtypes were expressed from P0 on nerve cell soma and qualitatively appeared to increase with age. Also DOR-IR and MOR-IR were detected at P8 and P16 in nerve fibers within the spiral ganglion. In the limbus DOR-IR was detected at P8 and P16 on cells proximal to the tectorial membrane while MOR-IR was detected more distally. In general these findings demonstrate that within the inner ear each receptor subtype follows specific temporal and spatial developmental patterns, some of which may be associated to the onset of hearing. The data provide further evidence that the opioid system may play a role in the development and functioning of the inner ear. PMID- 14553899 TI - Disappearance of click-evoked potentials on the neck of the guinea pig by pharmacological and surgical destruction of the peripheral vestibular afferent system. AB - In order to establish an animal model of acoustically evoked vestibulo-collic reflex, the so-called vestibular evoked myogenic potential in humans, potentials evoked by loud clicks on the neck of the guinea pig were recorded using subjects whose peripheral vestibular endorgans or vestibular afferents had been damaged. Four normal control guinea pigs, four guinea pigs that received an intramuscular injection of gentamicin for 20 days (90 mg/kg/day) and five guinea pigs whose vestibular nerves were surgically sectioned were used in this study. Under general anesthesia with an intraperitoneal injection of pentobarbital sodium (40 mg/kg), auditory brainstem responses (ABRs) were recorded. Then, potentials evoked by loud clicks on the pre-vertebral muscle at the level of the third cervical vertebral bone were recorded using silver ball electrodes. As a result, a distinctive negative peak (NP) with a latency of 6-8 ms was recorded in all animals in the control group. NP was not observed in the gentamicin-administered group while ABR was preserved. After sectioning the vestibular nerve, NP was abolished while ABR was preserved. From these results, NP could be of vestibular origin. These results are in agreement with a previous report of NP using subjects whose cochlea had been damaged pharmacologically. PMID- 14553900 TI - Distribution of catestatin-like immunoreactivity in the human auditory system. AB - Chromogranin A (CgA) belongs to the family of chromogranin peptides which are contained in large dense-core vesicles. The novel CgA fragment catestatin (bovine CgA(344-364); RSMRLSFRARGYGFRGPGLQL) is a potent inhibitor of catecholamine release by acting as a nicotinic cholinergic antagonist. Catestatin is a recently characterized neuropeptide, consisting of 21 amino acids, which might play an autocrine regulatory role in neuroendocrine secretion through its interaction with different nicotinic acetylcholine receptor subtypes. This study investigates for the first time the distribution of this peptide in the human auditory system using immunohistochemistry. A high density of catestatin-like immunoreactivity (catestatin-LI) is located in the spiral ganglion cells. In the dorsal cochlear nucleus, a high density of catestatin-LI consists of varicose fibers, immunoreactive varicosities and immunoreactive neurons. A moderate density is detected in the ventral cochlear and the medial vestibular nucleus. A low density is found in the inferior colliculus and superior olivary complex. The study indicates that catestatin is distinctly distributed in the auditory system, suggesting a role as a neuromodulatory peptide. Further studies should elucidate a possible interaction with other neurotransmitters in the auditory system. PMID- 14553901 TI - Expression of prestin, a membrane motor protein, in the mammalian auditory and vestibular periphery. AB - Hair cells are specialized mechanoreceptors common to auditory and vestibular sensory organs of mammalian and non-mammalian species. Different hair cells are believed to share common features related to their mechanosensory function. It has been shown that hair cells possess various forms of motile properties that enhance their receptor function. Membrane-based electromotility is a form of hair cell motility observed in isolated outer hair cells (OHCs) of the cochlea. A novel membrane protein, prestin, recently cloned from gerbil and rat tissues, is presumably responsible for electromotility. We cloned prestin from mouse organ of Corti and confirmed strong homology of this protein among different rodent species. We explored whether or not prestin is present in hair cells of the vestibular system. Using reverse transcription-polymerase chain reaction, we demonstrated that prestin is expressed in mouse and rat auditory and vestibular organs, but not in chicken auditory periphery. In situ hybridization and immunolocalization studies confirmed the presence of prestin in OHCs as well as in vestibular hair cells (VHCs) of rodent saccule, utricle and crista ampullaris. However, in the VHCs, staining of varying intensity with anti-prestin antibodies was observed in the cytoplasm, but not in the lateral plasma membrane or in the stereociliary membrane. Whole-cell patch-clamp recordings showed that VHCs do not possess the voltage-dependent capacitance associated with membrane-based electromotility. We conclude that although prestin is expressed in VHCs, it is unlikely that it supports the form of somatic motility observed in OHCs. PMID- 14553902 TI - Learning in discrimination of frequency or modulation rate: generalization to fundamental frequency discrimination. AB - Fifteen initially inexperienced subjects were trained for 4 weeks (12 2-h sessions) in frequency discrimination with pure tones around 88, 250, or 1605 Hz, or amplitude modulation rate discrimination of noise bands, using modulation rates around 88 or 250 Hz. Before, in the middle of, and after this training period, pure-tone frequency discrimination thresholds (DLFs), harmonic complex tone fundamental frequency discrimination thresholds (DLF0s), and amplitude modulation rate discrimination thresholds (DLFMs) were measured in several conditions including the trained one. Training in pure-tone frequency discrimination resulted in significantly larger improvements in DLF0s when the test complexes contained resolved harmonics than when they were composed of unresolved harmonics. This result supports the hypothesis that the discrimination of the F0 of resolved harmonics shares common underlying mechanisms with the frequency discrimination of pure tones. Training in rate discrimination did not result in larger DLF0 improvements for unresolved than for resolved harmonics. PMID- 14553903 TI - Thapsigargin suppresses cochlear potentials and DPOAEs and is toxic to hair cells. AB - Thapsigargin, a drug that inhibits sarco-endoplasmic reticulum Ca(2+) ATPases (SERCAs), was infused into the perilymph compartment of the guinea pig cochlea in increasing concentrations (0.1-10 microM) while sound evoked cochlear potentials were monitored. Thapsigargin significantly suppressed the compound action potential of the auditory nerve, cochlear microphonics, and increased N(1) latency at low (56 dB SPL) and high intensity (92 dB SPL) levels of sound, suppressed low intensity sound evoked summating potential (SP) and greatly increased the magnitude of the high intensity sound evoked SP. At 10 microM, the drug suppressed the cubic distortion product otoacoustic emissions (2f(1)-f(2)=8 kHz, f(2)=12 kHz) evoked by both high and low intensity primaries (45, 60, 70 dB SPL). Thapsigargin (10 microM; 30 min) increased the endocochlear potential slightly (5 mV). In chronic animals, thapsigargin (10 microM; 60 min) destroyed many outer hair cells and some inner hair cells, especially in the basal turns. These effects are consistent with the hypothesis that the inhibition of the SERCAs affects the function of the cochlear amplifier and outer hair cells to a greater degree than it affects other functions of the cochlea. PMID- 14553904 TI - Sound-evoked efferent effects on cochlear mechanics of the mustached bat. AB - The influence of the crossed medial efferent system on cochlear mechanics of the mustached bat was tested by measuring delayed evoked otoacoustic emissions (DEOAEs), cochlear microphonics, distortion product otoacoustic emissions (DPOAEs) and stimulus frequency otoacoustic emissions. Contralaterally delivered sinusoids, broadband noise and bat echolocation calls were used for acoustic stimulation of the efferent system. With all four measures we found a level dependent suppression under stimulation with both broadband noise and echolocation calls. In addition, the sharply tuned cochlear resonance of the mustached bat which is involved in processing echolocation signals at 61 kHz shifted upward in frequency by several 100 Hz. Presentation of sinusoids did not have any significant effect. DEOAEs and DPOAEs were in some cases enhanced during contralateral presentation of the bat calls at moderate intensities. The most important function of the efferent system in the mustached bat might be the control of the extraordinarily fine-tuned resonator of this species, which is close to instability as evident from the very pronounced evoked otoacoustic emissions which sometimes convert into spontaneous otoacoustic emissions of high level. PMID- 14553905 TI - C-Fos immunoreactivity mapping of the auditory system after electrical stimulation of the cochlear nerve in rats. AB - The aim of this study was to establish the use of c-Fos immunoreactivity as a marker for functional mapping in the auditory system in response to direct electrical stimulation of the cochlear nerve in the cerebellopontine angle. In rats the cochlear nerve was electrically stimulated with a biphasic current (120 250 microA, 5 Hz) for 30 min using a bipolar concentric Tungsten electrode. Bilateral cochlectomy was performed in a control group in order to investigate basal expression of c-Fos in the auditory brainstem nuclei. The response of auditory brainstem nuclei to electrical stimulation and the completeness of cochlear ablations were electrophysiologically verified. After the experiments, the animals were prepared for cryotomy and c-Fos immunohistochemistry. The results were morphologically analyzed and statistically compared among groups. In anesthetized animals with unilateral electrical stimulation of the cochlear nerve increased expression of c-Fos was detected in the ipsilateral ventral (VCN) and bilateral dorsal cochlear nucleus (DCN), whereas the VCN of the contralateral side revealed only few immunoreactive cells. In animals with bilateral cochlear ablation the number of c-Fos reactive cell nuclei representing basal expression was generally low in the VCN and DCN of both sides. Our data show that electrical stimulation of the cochlear nerve leads to increased expression of c-Fos in the cochlear nucleus. It also confirms bilateral connections between the cochlear nuclei. These experimental results suggest that c-Fos immunoreactivity mapping provides a powerful tool for functional investigations on the cellular level after direct electrical stimulation of the cochlear nerve. Future functional studies analyzing the effect of electrical stimulation of the central auditory system as performed by auditory brainstem implants could be investigated in detail by mapping c-Fos expression on cellular level. PMID- 14553906 TI - Enhancement of acoustic evoked potentials and impairment of startle reflex induced by reduction of GABAergic control of the neural substrates of aversion in the inferior colliculus. AB - The neural network of the inferior colliculus (IC), implicated in the generation of defensive behavior to aversive acoustic stimuli, is under tonic GABAergic control. Dopamine also seems to have a modulatory role in these neural circuits. It is still unclear how such changes in transmission of acoustic information influence the motor expression of the defensive behavior. Startle reaction to a sudden noise has been used as an effective way to measure the motor reactivity of rats to fearful acoustic stimuli. In this work we examined the processing of sensorial information--assessed by the recording of auditory evoked potentials (AEP)--and the behavioral effects--evaluated by the freezing and startle responses--during the reduction of GABA levels caused by microinjections of semicarbazide (SMC, 6 microg/0.2 microl), a glutamic acid decarboxylase inhibitor, into the IC. These data were compared to the effects of the overall arousal elicited by apomorphine (APO, 0.5 mg/kg, i.p.). The results obtained show that IC microinjections of SMC induced freezing behavior, enhanced the AEP and impaired the startle reaction to a loud sound. On the other hand, APO changed neither the AEP nor the startle in the same experimental conditions. These results suggest that the release of GABAergic control of the neural substrates of aversion in the IC results in an increased processing of auditory information along with an inhibitory influence on the motor pathways responsible for the startle response. PMID- 14553908 TI - The effect of L-carnitine on the prevention of experimentally induced myringosclerosis in rats. AB - The objective of this study was to investigate the possible effect of L-carnitine on the prevention of experimentally induced myringosclerosis. Twenty Sprague Dawley rats were bilaterally myringotomized. The rats were divided into two groups randomly: group 1 which were intraperitoneally administered saline and group 2 which were intraperitoneally administered L-carnitine. Blood samples were collected for biochemical evaluation and the tympanic membranes were harvested after 28 days. Histopathological and immunohistochemical evaluation were done under light microscopy. The mean malondialdehyde levels were 3.9+/-0.9 in group 2, and 7.9+/-1.1 in group 1 (P<0.001), nitric oxide levels were 25.6+/-6.4 in group 2 and 30.8+/-8.2 in group 1 (P=0.14) and acetylcholinesterase was 1035+/-60 in group 2 and 678+/-35 in group 1 (P=0.001). Myringosclerosis was more frequent and severe in group 1 than group 2 (P<0.007). Immunoreactivity was seen in 16 of 20 tympanic membranes in group 2 and six of 20 tympanic membranes in group 1 (P=0.005). We conclude that L-carnitine diminishes the occurrence of myringosclerosis in rats after myringotomy possibly by antioxidant activity and decreasing the formation of reactive oxygen species. PMID- 14553907 TI - Corticofugal modulation of amplitude domain processing in the midbrain of the big brown bat, Eptesicus fuscus. AB - Recent studies have shown that the corticofugal system systematically modulates and improves subcortical signal processing in the frequency, time and spatial domains. The present study examined corticofugal modulation of amplitude sensitivity of 113 corticofugally inhibited neurons in the central nucleus of the inferior colliculus (IC) of the big brown bat, Eptesicus fuscus. Cortical electrical stimulation decreased the number of impulses and increased the response latency of these neurons. They had an average of 5.9+/-4.4 kHz best frequency (BF) differences between collicular and electrically stimulated cortical neurons. Cortical electrical stimulation synchronized with sound stimulation for 30 min compressed the rate-amplitude functions of half (56, 49.6%) of these collicular neurons and shifted their minimum thresholds (MT) and dynamic ranges (DR) toward that of electrically stimulated cortical neurons for as long as 40 min. These collicular neurons had an average of 1.6+/-1.4 kHz BF differences. The shift in collicular MT and DR significantly increased with differences in MT and DR between collicular and cortical neurons. Cortical electrical stimulation also shifted the BF and best amplitude (BA) of collicular neurons toward that of cortical neurons. The BF shift increased with BF differences and the BA shift increased with BA differences. These data suggest that the corticofugal system modulates collicular responses on the basis of topographic projections between the IC and auditory cortex. However, corticofugal modulation of collicular amplitude sensitivity is primarily dependent upon the difference but not the absolute amplitude sensitivity between collicular and cortical neurons. PMID- 14553909 TI - Hearing in American leaf-nosed bats. III: Artibeus jamaicensis. AB - We determined the audiogram of the Jamaican fruit-eating bat (Phyllostomidae: Artibeus jamaicensis), a relatively large (40-50 g) species that, like other phyllostomids, uses low-intensity echolocation calls. A conditioned suppression/avoidance procedure with a fruit juice reward was used for testing. At 60 dB SPL the hearing range of A. jamaicensis extends from 2.8 to 131 kHz, with an average best sensitivity of 8.5 dB SPL at 16 kHz. Although their echolocation calls are low-intensity, the absolute sensitivity of A. jamaicensis and other 'whispering' bats does not differ from that of other mammals, including other bats. The high-frequency hearing of A. jamaicensis and other Microchiroptera is slightly higher than expected on the basis of selective pressure for passive sound localization. Analysis suggests that the evolution of echolocation may have been accompanied by the extension of their high-frequency hearing by an average of one-half octave. With respect to low-frequency hearing, all bats tested so far belong to the group of mammals with poor low-frequency hearing, i.e., those unable to hear below 500 Hz. PMID- 14553910 TI - The order of testing effect in otoacoustic emissions and its consequences for sex and ear differences in neonates. AB - The amplitude values of transient-evoked otoacoustic emissions, recorded from a large sample of neonates, were used to examine the asymmetry between ears tested and the differences due to the sex of the subject. Whilst the sex difference, with females having larger responses than males, has been a consistent finding in previous reports, the right/left ear difference, with the right ear giving a larger response than the left, has produced variable results that differed between laboratories. In this study, the sex difference was confirmed with females giving a 1.2 dB greater response than males. It was not affected by the age of the neonate. A significant effect of test order was found. The measured right/left difference was enhanced when the right ear was tested first but was diminished when the left ear was tested first. If the left ear is tested first then the measured right/left difference would be about 0.5 dB whereas, if the right ear is tested first, the measured right/left difference would be about 1.5 dB. When male/female comparisons were made for right and left ears separately and for the same ear tested first, the sex differences were the same for all four conditions. The sex and right/left differences have been confirmed as statistically significant effects and the order effect could explain the discrepancies and variability of the right/left differences reported in the literature. PMID- 14553911 TI - Taurine: new implications for an old amino acid. AB - Taurine is a semi-essential amino acid and is not incorporated into proteins. In mammalian tissues, taurine is ubiquitous and is the most abundant free amino acid in the heart, retina, skeletal muscle, brain, and leukocytes. In fact, taurine reaches up to 50 mM concentration in leukocytes. Taurine has been shown to be tissue-protective in many models of oxidant-induced injury. One possibility is that taurine reacts with hypochlorous acid, produced by the myeloperoxidase pathway, to produce the more stable but less toxic taurine chloramine (Tau-Cl). However, data from several laboratories demonstrate that Tau-Cl is a powerful regulator of inflammation. Specifically, Tau-Cl has been shown to down-regulate the production of pro-inflammatory mediators in both rodent and human leukocytes. Taurolidine, a derivative of taurine, is commonly used in Europe as an adjunctive therapy for various infections as well as for tumor therapy. Recent molecular studies on the function of taurine provide evidence that taurine is a constituent of biologic macromolecules. Specifically, two novel taurine-containing modified uridines have been found in both human and bovine mitochondria. Studies investigating the mechanism of action of Tau-Cl have shown that it inhibits the activation of NF-kappaB, a potent signal transducer for inflammatory cytokines, by oxidation of IkappaB-alpha at Met45. Key enzymes for taurine biosynthesis have recently been cloned. Cysteine sulfinic acid decarboxylase, a rate-limiting enzyme for taurine biosynthesis, has been cloned and sequenced in the mouse, rat and human. Another key enzyme for cysteine metabolism, cysteine dioxygenase (CDO), has also been cloned from rat liver. CDO has a critical role in determining the flux of cysteine between cysteine catabolism/taurine synthesis and glutathione synthesis. Taurine transporter knockout mice show reduced taurine, reduced fertility, and loss of vision due to severe apoptotic retinal degeneration. Apoptosis induced by amino chloramines is a current and important finding since oxidants derived from leukocytes play a key role in killing pathogens. The fundamental importance of taurine in adaptive and acquired immunity will be unveiled using genetic manipulation. PMID- 14553912 TI - Susceptibility of hematopoietic stem cells to pathogens: role in virus/bacteria tropism and pathogenesis. AB - Human hematopoietic stem cells (HSCs) are generated in the bone marrow and differentiate into erythrocytes, granulocytes, monocytes, megacaryocytes, and lymphocytes. HSCs may be manipulated under different conditions. Advances in cell biology result in a better understanding of the relationship between viruses/bacteria and hematopoietic cells. Microbial infections can lead to profound disturbance of hematopoiesis. Infection may augment the production of cytokines, with proliferation and differentiation of the stem cells. Alternatively, infection may lead to destruction of progenitor cells. This results in defective hematopoiesis in certain infections. Since circulating CD34+ cells represent a distinct progenitor pool responsible for seeding extramedullary sites of hematopoiesis, infected peripheral blood-derived CD34+ progenitor cells may serve to disseminate pathogens into diverse anatomic sites. Therefore, progenitor cell infection may additionally effect long-term functional consequences within extramedullary sites of lymphopoiesis. A variety of viruses have been reported to target HSCs, whereas quiescent human HSCs are fully resistant to infection by different bacteria. For susceptibility of HSCs to infectious agents pathogen-receptor interaction plays an important role in virus/bacteria tropism and pathogenesis. PMID- 14553913 TI - Localizing cell division in spherical Escherichia coli by nucleoid occlusion. AB - The spatial relationship between FtsZ localization and nucleoid segregation was followed in Escherichia coli thyA cells, made spheroidal by brief exposure to mecillinam and after manipulating chromosome replication time using changes ('steps') in thymine concentration [Zaritsky et al., Microbiology 145 (1999) 1015 1022]. In such cells, fluorescent FtsZ-GFP arcs did not overlap the DAPI-stained nucleoids. It is concluded that FtsZ rings are deposited between segregating nucleoids, consistent with the nucleoid occlusion model [Woldringh et al., J. Bacteriol. 176 (1994) 6030-6038]. PMID- 14553914 TI - Molecular cloning and characterization of a novel leptospiral lipoprotein with OmpA domain. AB - A novel antigenic protein of pathogenic Leptospira, Loa22, was identified by using the PhoA fusion method followed by immunoblotting with convalescent mouse sera. Loa22 was shown to be a lipoprotein having a C-terminal OmpA consensus domain. Loa22 was detected among pathogenic leptospires but not among non pathogenic leptospires, suggesting the possible involvement of this protein in virulence. The results of three different experiments suggested that Loa22 is located in the outer membrane and a small portion is exposed on the cell surface. Thus, Loa22 may be a candidate for a novel vaccine against infection with pathogenic leptospires. PMID- 14553915 TI - Different contributions of the outer and inner R-core residues of lipopolysaccharide to the recognition by spike H and G proteins of bacteriophage phiX174. AB - The binding of spike H and G proteins of bacteriophage phiX174 with lipopolysaccharides (LPSs) were evaluated by a competitive enzyme-linked plate assay using the biotin-labeled LPS of Escherichia coli C, one of a host strain, and the non-labeled LPSs having different R-core polysaccharide lengths. H protein promptly decreased its affinity when some saccharide residues were truncated from the outer R-core. However, G protein showed significant affinity to the LPSs lacking all the residues of the outer R-core and some of the inner R core. Thus, G protein rather than H protein well recognized the residues of the inner R-core of LPS. PMID- 14553916 TI - Expression of SA5K, a secretion antigen of Mycobacterium tuberculosis, inside human macrophages and in sputum from tuberculosis patients. AB - An 8.3 kDa protein (SA5K), secreted by Mycobacterium tuberculosis/Mycobacterium bovis bacillus Calmette-Guerin (BCG) in culture filtrate, has been previously described in our laboratory. In the present study, analysis of the distribution of SA5K gene (Rv1174c) among M. tuberculosis strains, isolated from a wide variety of clinical specimens, revealed that the gene is present in all clinical isolates analyzed (29/29). SA5K expression inside human macrophages infected with BCG was demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) on RNA extracted from bacterial cells following 24 and 48 h of infection. In addition, in order to evaluate whether SA5K gene was also expressed at the site of infection in the lung, a nested RT-PCR assay was developed to detect specific mRNA in sputum samples collected from smear positive tuberculosis patients. SA5K mRNA was detected in all the samples containing high numbers of tubercle bacilli demonstrating that the corresponding gene is expressed during the course of clinical infection. PMID- 14553917 TI - Study of 18S rRNA and rDNA stability by real-time RT-PCR in heat-inactivated Cryptosporidium parvum oocysts. AB - The public health problem posed by Cryptosporidium parvum has led the water supply industry to develop analytical tools for detecting viable oocysts in water. In this study, we report on a TaqMan real-time reverse transcription polymerase chain reaction (RT-PCR) method that targets and quantifies C. parvum 18S rRNA. To study the suitability of 18S rRNA as an indicator of Cryptosporidium oocyst viability, the stability of 18S rRNA and rDNA was monitored by real-time RT-PCR following various Cryptosporidium heat treatments. Decay of 18S rRNA was first observed after a 20-min treatment of C. parvum oocysts at 95 degrees C and was still detectable after 4 h. In contrast, rDNA was more heat resistant. The stability of 18S rRNA and rDNA was also studied after oocyst lysis by thermal shocks in the presence and absence of Chelex-100. In the former case, both rRNA and rDNA were degraded whereas in the presence of Chelex-100 both molecules were protected from heat degradation and were still detected after 4 h at 95 degrees C following thermal shocks. Our results indicate that 18S rRNA detection may not be directly associated with viability following heat inactivation of Cryptosporidium oocysts even if in all the experiments 18S rRNA was less stable than rDNA. PMID- 14553918 TI - beta-Lactam induction of colanic acid gene expression in Escherichia coli. AB - An unexpected observation led us to examine the relationship between beta-lactam exposure and synthesis of colonic acid capsular polysaccharide in Escherichia coli. Strains containing a cps-lacZ transcriptional fusion were challenged with antibiotics having various modes of action, and gene expression was detected by a disk-diffusion assay and in broth cultures. The cps genes were induced by a subset of beta-lactams but not by agents inhibiting protein synthesis or DNA replication, indicating that cps expression was specific and not due to stresses accompanying cell death or by a general inhibition of peptidoglycan synthesis. A narrow concentration just below the MIC triggered cps expression in liquid culture, suggesting the response may be triggered by near-lethal levels of antibiotic. Because colanic acid is important for maturation of biofilm architecture, antibiotics that increase its synthesis might exacerbate the formation or persistence of biofilms. PMID- 14553919 TI - Tn5060 from the Siberian permafrost is most closely related to the ancestor of Tn21 prior to integron acquisition. AB - A Tn21-related mercury resistance transposon, Tn5060, has been isolated from Pseudomonas sp. strain A19-1 from a 8,000-10,000-year-old Siberian permafrost sample, and sequenced. Like Tn21, the element transposes to different plasmids at a frequency of 10(-2)-10(-3) per target plasmid transfer. Comparison of the complete Tn5060 DNA sequence (8,667 bp) with that of Tn21 (19,672 bp) shows that Tn5060 does not contain integron In2 and deviates from Tn21 in four nucleotide positions. These and other comparative data demonstrate that Tn5060 is the most closely related of the characterized mercury resistances to the as yet hypothetical immediate ancestor of Tn21, TnX. PMID- 14553920 TI - Phylogenetic affinity of a Giardia lamblia cysteine desulfurase conforms to canonical pattern of mitochondrial ancestry. AB - Among a few potential archezoan groups, only the Metamonada (diplomonads, retortamonads, and oxymonads) still retain the status of amitochondriate protists that diverged before the acquisition or retention of mitochondria. Indeed, finding that diplomonad genomes harbor a gene encoding a mitochondrial type chaperonin 60, the most compelling evidence for their secondarily amitochondriate nature, may be interpreted as an acquisition of this important general chaperone during some transient alpha-proteobacterial endosymbiosis. Recently published data on the cysteine desulfurase IscS demonstrated an alpha-proteobacterial origin of mitochondrial enzymes including a diplomonad Giardia lamblia homolog. An extended phylogenetic analysis of IscS is reported here that revealed a full canonical pattern of mitochondrial ancestry for the giardial enzyme. The above canonical pattern, a sister group relationship of mitochondria and rickettsiae exclusive of free-living alpha-proteobacteria, was robustly confirmed by a comprehensive analysis of Cob and Cox1 subunits of the respiratory chain encoded by resident mitochondrial genes. Given that Fe-S cluster assembly involving IscS represents an essential mitochondrial function, these data strongly suggest that diplomonads once harbored bona fide mitochondria. PMID- 14553921 TI - Binding of Porphyromonas gingivalis fimbriae to Treponema denticola dentilisin. AB - Treponema denticola has been reported to coaggregate with Porphyromonas gingivalis and localize closely together in matured subgingival plaque. In this study of the interaction of T. denticola with P. gingivalis, the P. gingivalis fimbria-binding protein of T. denticola was identified by two-dimensional electrophoresis followed by a ligand overlay assay with P. gingivalis fimbriae, and was determined to be dentilisin, a chymotrypsin-like proteinase of T. denticola. The binding was further demonstrated with a ligand overlay assay using an isolated GST fusion dentilisin construct. Our results suggest that P. gingivalis fimbriae and T. denticola dentilisin are implicated in the coaggregation of these bacteria. PMID- 14553922 TI - Purification and properties of a new psychrophilic metalloprotease (Fpp2) in the fish pathogen Flavobacterium psychrophilum. AB - To go further into the characterization of the proteolysis exocellular system of the salmonid pathogen Flavobacterium psychrophilum, the purification and characterization of a novel protease designated Fpp2 (F. psychrophilum protease 2) was undertaken. A protease (Fpp2) hydrolyzing azocasein was purified. The Fpp2 can be defined as a metalloprotease, it had an estimated molecular mass of 62 kDa with calcium playing an important role in the thermostability of the enzyme. Proteolytic activity was optimal at pH 6.0-7.0 and 24 degrees C and activation energy for the hydrolysis of azocasein was determined to be 5.4 kcal mol(-1), being inactive at temperatures above 42 degrees C. All these results are characteristic of 'cold adapted enzymes'. Fpp2 proved to be a broad range hydrolytic enzyme because in optimal conditions it was able to hydrolyze matrix and muscular proteins. It can be concluded that the Fpp1, a previously characterized 55 kDa metalloprotease, and the Fpp2 protease were produced under different physiological conditions and were immunologically as well as biochemically different. PMID- 14553923 TI - Identification of tdh-positive Vibrio parahaemolyticus from an outbreak associated with raw oyster consumption in Spain. AB - Between August and September 1999, a total of 64 cases of illness were identified in three episodes of acute gastroenteritis associated with the consumption of live oysters from a typical outdoor street market in Galicia (northwest Spain). Nine case patients were hospitalized and analysis of their stool samples revealed the presence of Vibrio parahaemolyticus. The strains isolated from two stool samples were studied for antibiotic susceptibility, biochemical characteristics and presence of virulence factors. Both isolates were Kanagawa phenomenon positive and produced thermostable direct hemolysin, which is related to pathogenicity in humans. These results show the presence of pathogenic V. parahaemolyticus in mollusks harvested in Europe and reveal the risk of illness associated with their consumption, suggesting the revision of V. parahaemolyticus risk assessment associated with consumption of raw live shellfish. PMID- 14553924 TI - Isolation, characterization and expression of a cyclin from Leishmania donovani. AB - We have cloned and sequenced a DNA fragment (approximately 1 kb) containing a complete open reading frame from a cDNA library of Leishmania donovani promastigotes. The alignment of the derived polypeptide sequence and the modeling studies revealed that the protein is highly homologous to the mammalian cyclins having conserved cyclin box and substrate-docking motif. Northern blot analysis of the RNA isolated from synchronized L. donovani promastigotes showed periodic expression of the message with maximum abundance at S-phase suggesting its involvement in the events related to the regulation of DNA replication. The results confirm that we have isolated a cyclin molecule from L. donovani (LdCyc1) which may play an important role in the regulation of the parasite cell cycle. PMID- 14553925 TI - Identification of rpoE and nadB as host responsive elements of Yersinia enterocolitica. AB - We describe the identification of the Yersinia enterocolitica rpoE gene, which encodes the alternative sigma factor sigmaE, and the divergently transcribed nadB gene, as genes that are expressed during infection of mice. As in Escherichia coli, rpoE of Y. enterocolitica is essential for growth and its expression is autoregulated. Despite the similarities of the rpoE operons of Y. enterocolitica and E. coli, there are considerable differences in the response to extracytoplasmic stress. Unlike in E. coli, sigmaE of Y. enterocolitica is not induced by heat shock or ethanol. Overproduction of the outer membrane protein Ail does not lead to the induction of rpoE expression. However, rpoE expression is induced by osmotic stress. PMID- 14553926 TI - Use of rpoB sequences for phylogenetic study of Mycoplasma species. AB - rpoB sequences encoding the beta-subunit of RNA polymerase were determined in 26 Mycoplasma species for phylogenetic study. The portion of rpoB DNA used in this study showed a high degree of variation in terms of size and sequence among species. The rpoB phylogenies inferred from amino acid and nucleotide sequences were used to divide the mycoplasmas into two groups, a 'pneumoniae group' and a 'hominis group', which was consistent with the result from 16S rDNA sequence analysis. However, phylogenetic relationships within these groups differed in the two gene trees, which were supported by the incongruence length difference (ILD) test. This indicates that multiple gene sequences must be applied to infer accurate phylogenetic relationships among the mycoplasmas. The rpoB sequence, and especially the deduced amino acid sequence, offers a good alternative marker. PMID- 14553927 TI - Transcriptional analysis of the rpoE gene encoding extracytoplasmic stress response sigma factor sigmaE in Salmonella enterica serovar Typhimurium. AB - The rpoE gene of Salmonella enterica serovar Typhimurium (S. Typhimurium), which encodes the extracytoplasmic stress response sigma factor sigmaE, is critically important for the virulence of S. Typhimurium. We analysed expression of rpoE by wild-type and mutant bacteria grown in different conditions by S1-nuclease mapping using RNA, and using in vivo reporter gene fusions. Three promoters, rpoEp1, rpoEp2 and rpoEp3, were located upstream of the S. Typhimurium rpoE gene. The promoters were differentially expressed during growth and under several stress conditions including cold shock. Expression from the rpoEp3 promoter was absent in an S. Typhimurium rpoE mutant, demonstrating its dependence upon sigmaE. The level of mRNA corresponding to rpoEp3 was also higher in a cpxR mutant, indicating a negative regulation of the promoter by the Cpx system. Using this rpoE-dependent promoter, we optimised a two-plasmid system for identification of promoters recognised by S. Typhimurium sigmaE. The rpoEp3 promoter was active in the Escherichia coli two-plasmid system and has an identical transcription start point as in S. Typhimurium but only after induction of S. Typhimurium rpoE expression. PMID- 14553928 TI - Functional analysis of Ras in Colletotrichum trifolii. AB - Ras is a small monomeric GTP binding protein that transduces signals for growth and differentiation of eukaryotic organisms. Previously, a unique ras gene, designated Ct-ras, was cloned from the alfalfa fungal phytopathogen, Colletotrichum trifolii. Expression of Ct-Ras in mouse fibroblast cells (NIH3T3) demonstrated that Ct-ras is functionally similar to the mammalian ras genes since activating mutations of Ct-ras caused oncogenic phenotypes in nu/nu mice, including tumors. In C. trifolii, activated 'oncogenic' Ras (Val2) induced abnormal hyphal proliferation, defects in polarized growth and significantly reduced differentiation such as conidiation and appressorium formation in a nutrient dependent manner. Gene disruption of ct-ras was lethal. To further evaluate the function of Ct-Ras in C. trifolii, three different approaches were used: overexpression of cytosolic Ras by CAAX box deletion; expression of dominant negative Ct-RasT22N; and antisense ct-ras expression. Results showed that suppression of Ct-Ras activity significantly decreases fungal germination frequencies and hyphal growth rates. Taken together, these data suggest involvement of Ct-Ras in regulation of fungal cell growth and differentiation. PMID- 14553930 TI - Aerobic and anaerobic nitrate and nitrite reduction in free-living cells of Bradyrhizobium sp. (Lupinus). AB - Induction, energy gain, effect on growth, and interaction of nitrate and nitrite reduction of Bradyrhizobium sp. (Lupinus) USDA 3045 were characterized. Both nitrate and nitrite were reduced in air, although nitrite reduction was insensitive to ammonium inhibition. Anaerobic reduction of both ions was shown to be linked with energy conservation. A dissimilatory ammonification process was detected, which has not been reported in rhizobia so far. Nevertheless, anaerobic conversion of nitrate to ammonium was lower than 40%, which suggests the presence of an additional, nitrite reductase of denitrifying type. Nitrite toxicity caused a non-linear relationship between biomass produced and >2 mM concentrations of each N oxyanion consumed. At > or =5 mM initial concentrations of nitrate, a stoichiometric nitrite accumulation occurred and nitrite remained in the medium. This suggests an inhibition of nitrite reductase activity by nitrate, presumably due to competition with nitrate reductase for electron donors. Lowering of growth temperature almost completely diminished nitrite accumulation and enabled consumption as high as 10 mM nitrate, which confirms such a conclusion. PMID- 14553929 TI - csp-like genes of Lactobacillus delbrueckii ssp. bulgaricus and their response to cold shock. AB - The two csp-like genes from the lactic acid bacterium Lactobacillus delbrueckii ssp. bulgaricus were characterized and designated cspA and cspB. The gene cspA has been identified using a polymerase chain reaction (PCR)-based approach with degenerated primers and further characterized using an inverse PCR strategy. cspA encodes a protein of 65 amino acid residues which displays between 81 and 77% identity with proteins CspL and CspP of Lactobacillus plantarum. cspB has been identified as a cspA ortholog using the partial sequence of the L. bulgaricus ATCC11842. cspB encodes a protein of 69 amino acids which has 42% identity with CspA. Northern blot analyses showed that cspA is transcribed as a single gene and that its transcription increased after a temperature downshift from 42 to 25 degrees C. In contrast, cspB is part of an operon transcribed at constant level irrespective of the temperature. These results indicate that cspA encodes the only Csp-like protein of L. bulgaricus induced by a downshift of temperature. PMID- 14553931 TI - Transcriptional enhancement of RT-PCR for rapid and sensitive detection of Noroviruses. AB - Previously reported nucleic acid sequence-based amplification (NASBA) primers specific for the GII Noroviruses were adapted for reverse transcriptase polymerase chain reaction (RT-PCR), and detection sensitivity was then enhanced by a subsequent in vitro transcription of the RT-PCR amplicons. The NASBA-derived primers performed comparably to other broadly reactive GII Norovirus primers with respect to detection limits (i.e. 1 RT-PCR amplifiable unit (RT-PCRU) per reaction). Detection limits improved by approximately 1 log(10) to 0.3 RT-PCRU per reaction when transcriptional enhancement and electrochemiluminescence (ECL) hybridization followed RT-PCR. The method shows promise for improved detection sensitivity in instances where very low levels of virus contamination might be anticipated. PMID- 14553932 TI - Bacterial cell surface display for epitope mapping of hepatitis C virus core antigen. AB - Cell surface expression of protein has been widely used to display enzymes and antigens. Here we show that Pseudomonas syringae ice nucleation protein with a deletion of internal repeating domain (INC) can be used in Escherichia coli to display peptide in a conformationally active form on the outside of the folded protein by fusing to the C-terminus of INC. Diagnostic potential of this technology was demonstrated by effective mapping of antigenic epitopes derived from hepatitis C virus (HCV) core protein. Amino acids 1-38 and 26-53 of HCV core protein were found to react more sensitively in a native conformation with the HCV patient sera than commercial diagnostic antigen, c22p (amino acids 10-53) by display-ELISA. These results demonstrate that the bacterial cell surface display using INC is useful for peptide presentation and thus epitope mapping of antigen. PMID- 14553933 TI - The effects of Escherichia coli capsule, O-antigen, host neutrophils, and complement in a rat model of Gram-negative pneumonia. AB - Gram-negative enteric bacilli are agents of life-threatening pneumonia. The role of the bacterial capsule and O-antigen moiety of lipopolysaccharide in the pathogenesis of Gram-negative pneumonia was assessed. In a rat model of pneumonia the LD(50) of a wild-type extraintestinal pathogenic Escherichia coli strain (CP9) was significantly less than its isogenic derivatives deficient in capsule (CP9.137), O-antigen (CP921) or both capsule and O-antigen (CP923) (P< or =0.003). Studies using complement depleted or neutropenic animals established that both neutrophils and complement are important for the pulmonary clearance of E. coli. Data from these studies also support that capsule and O-antigen serve, at least in part, to counter the complement and neutrophil components of the pulmonary host defense response. Lastly, the contribution of E. coli versus neutrophils in causing lung injury was examined. Findings suggest that E. coli virulence factors and/or non-neutrophil host factors are more important mediators of lung injury than neutrophils. These findings extend our understanding of Gram negative pneumonia and have treatment implications. PMID- 14553934 TI - Acidic pH is required for the functional assembly of the type III secretion system encoded by Salmonella pathogenicity island 2. AB - Salmonella enterica employs two type III secretion systems (T3SS) for interactions with host cells during pathogenesis. The T3SS encoded by Salmonella pathogenicity island 2 (SPI2) is required for the intracellular replication of Salmonella and the survival inside phagocytes. During growth in vitro, acidic pH is a signal that promotes secretion of proteins by this T3SS. We analyzed protein levels and subcellular localization of various T3SS subunits under in vitro conditions at acidic or neutral pH, inducing or ablating secretion, respectively. Growth at acidic pH resulted in higher levels of SsaC, a protein forming the outer membrane secretin, without increasing expression of the operon containing ssaC. Acidic pH also induced oligomerization of SsaC subunits, a prerequisite for a functional secretin pore. It has previously been described that environmental stimuli resembling the intraphagosomal habitat of Salmonella control the expression of SPI2 genes. Here we propose that such stimuli also modulate the assembly of a functional T3SS that is capable of translocation of effector proteins into the host cell. PMID- 14553935 TI - Concurrence of losing a chromosome and the ability to produce destruxins in a mutant of Metarhizium anisopliae. AB - In a previous study, a spontaneous subtilisin pr1A and pr1B gene-deficient mutant of the entomopathogenic fungus Metarhizium anisopliae strain V275 has been identified [Wang, C.-S. et al. (2002) FEMS Microbiol. Lett. 213, 251-255]. The insecticidal metabolites of this mutant were studied further. High-performance liquid chromatography (HPLC) analysis indicated that the mutant isolate lost the ability to produce cyclic peptide toxins, destruxins, both in vitro and in vivo. Pulsed-field gel electrophoresis revealed that the mutant concurrently lost a 1.05 Mb (approximately) chromosome, demonstrating for the first time that a conditionally dispensable (CD) chromosome exists in the insect pathogenic fungus, M. anisopliae. Concurrence of losing the ability to produce destruxins and a CD chromosome in the mutant suggests that the toxin synthetase genes of M. anisopliae are located on this CD chromosome, as similarly described for plant pathogenic fungi. Semi-quantitative api ZYM analysis showed more biochemical disparities between the mutant and the wild-type strain. PMID- 14553936 TI - Effects of metronidazole and tinidazole on NTPDase1 and ecto-5'-nucleotidase from intact cells of Trichomonas vaginalis. AB - Here we report the effects of metronidazole and tinidazole on NTPDase1 and ecto 5'-nucleotidase from intact cells of Trichomonas vaginalis. Adenosine triphosphate (ATP) and adenosine diphosphate (ADP) hydrolysis was 5- to 7-fold higher for the fresh clinical strain, when compared with the ATCC (American Type Culture Collection) strain. ATP hydrolysis was activated in the presence of metronidazole in the ATCC strain, whilst it was inhibited 33% by 50 microM tinidazole in a fresh clinical isolate. The treatment of cells in the presence of metronidazole for 2 h inhibited ATP and ADP hydrolysis, whilst treatment with tinidazole inhibited ATP and ADP hydrolysis only in the fresh clinical isolate. The drugs did not change the ecto-5'-nucleotidase activity for both strains. Our results suggest that the modulation of extracellular ATP and ADP levels during treatment with these drugs could be a parasitic defence strategy as a survival mechanism in an adverse environment. PMID- 14553937 TI - Cloning and expression of a pectate lyase from the oral spirochete Treponema pectinovorum ATCC 33768. AB - The pelA gene, encoding a pectate lyase, from Treponema pectinovorum ATCC 33768 was isolated by heterologous expression of a cosmid library in Escherichia coli. In vitro transposon mutagenesis identified an open reading frame of 1293 bp capable of encoding a protein of 430 amino acids with a predicted amino-terminal signal sequence of 21 amino acids. Analysis of the amino acid sequence suggested that it is a member of the polysaccharide lyase family 10 of which all characterized members show pectate lyase activity. An amino-terminal His-tagged recombinant form of PelA was expressed and purified from E. coli. The recombinant enzyme has characteristics common to other bacterial pectate lyases such as an alkaline pH optimum, dependence on calcium ions for activity, and inhibition by zinc ions. PMID- 14553938 TI - Proteome analysis of Salmonella enterica serovar Typhimurium fis mutant. AB - Salmonella enterica serovar Typhimurium is an enteric pathogen and a principal cause of gastroenteritis in humans. The factor-for-inversion stimulation protein (Fis) is known to play a pivotal role in the expression of Salmonella pathogenicity island (SPI)-1 genes in addition to various cellular processes such as recombination, replication, and transcription. In order to understand Fis function in pathogenicity of Salmonella, we performed two-dimensional gel electrophoresis and identified proteins whose expression pattern is affected by Fis using mass spectrometry. The results revealed various proteins that can be grouped according to their respective cellular functions. These groups include the genes involved in the metabolism of sugar, flagella synthesis, translation, and SPI expression. Changes in SPI expression suggest the possibility that regulation of genes in SPI-2 as well as SPI-1 is affected by Fis. PMID- 14553940 TI - ATP-dependent glucokinase from the hyperthermophilic bacterium Thermotoga maritima represents an extremely thermophilic ROK glucokinase with high substrate specificity. AB - The gene (open reading frame (ORF) Tm1469, glk) encoding ATP-dependent ROK (repressors, ORFs, sugar kinases) glucokinase (ATP-GLK, EC 2.7.1.2) of the hyperthermophilic bacterium Thermotoga maritima was cloned and functionally expressed in Escherichia coli. The purified recombinant enzyme is a homodimer with an apparent molecular mass of 80 kDa composed of 36-kDa subunits. Rate dependence (at 80 degrees C) on glucose and ATP followed Michaelis-Menten kinetics with apparent Km values of 1.0 and 0.36 mM, respectively; apparent Vmax values were about 370 U mg(-1). The enzyme was highly specific for glucose as phosphoryl acceptor. Besides glucose only 2-deoxyglucose was phosphorylated to some extent, whereas mannose and fructose were not used. With a temperature optimum of 93 degrees C the enzyme is the most thermoactive bacterial ATP-GLK described. PMID- 14553939 TI - Purification, characterization, and physiological response of a catalase peroxidase in Mycobacterium sp. strain JC1 DSM 3803 grown on methanol. AB - A novel catalase-peroxidase (CP) from methanol-grown cells of Mycobacterium sp. strain JC1 was purified. The CP exhibited properties of both typical mycobacterial CPs (i.e. strict pH optimum, labile to heat treatment, capable of oxidizing NADH, and resistant to inhibition by 3-amino-1,2,4-triazole) and true catalases (i.e. stable against ethanol-chloroform treatment). The enzyme oxidized methanol and shared common antigenic groups with other mycobacteria. Isoniazid had almost no effect on the growth and expression of CP but inhibited the enzyme activity to some extent. Sodium nitroprusside arrested the growth but strongly stimulated the expression of CP with a concomitant increase in activity after the mid-exponential growth phase. PMID- 14553941 TI - DNA sequences of Mycobacterium leprae recovered from ancient bones. PMID- 14553942 TI - Changes in apoptosis-related pathways in acute myelocytic leukemia. AB - Expression analysis of apoptotic genes was performed for 15 patients with acute myelocytic leukemia (AML) at the time of diagnosis to identify genes and signaling pathways involved in the regulation of cell survival and apoptosis during leukemogenesis. cDNA array analysis revealed 34 genes whose expression was significantly different compared to others. Tumor suppressor genes TP53 and CDKN2A were downregulated and protooncogenes JUN and GRB10 were upregulated. Furthermore, several cellular signaling pathways acting either in cell cycle regulation or in apoptosis were altered. Deregulation was found in pathways that contribute to genomic stability (by downregulation of either TP53 or CSE1L and by upregulation of GADD45A) and regulate cell cycle progression (by downregulation of CDKN2A and upregulation of RBBP4, CDC37, and NEDD5). Alterations at the transcriptional level were identified, namely, upregulation of JUN and E2F5. Abnormalities were observed in the regulation of the caspases through upregulation of CASP8 and by altered expression of BCL2-related pathway. Extrinsic apoptotic signals mediated by IGFs were deregulated and the glutathione detoxification pathway was downregulated. These findings provide insight into the regulation of balance between apoptosis and cell proliferation signals, and suggest that these genes and pathways may have an important role in the pathogenesis of AML. PMID- 14553943 TI - Newly established Askin tumor cell line and overexpression of focal adhesion kinase in Ewing sarcoma family of tumors cell lines. AB - Askin tumor is a malignant small round cell tumor that originates from the thoracopulmonary region and is a member of Ewing sarcoma family of tumors (ESFT). Only a few Askin tumor cell lines have been established. An Askin tumor cell line, designated MP-ASKIN-SA, was established from the left thoracic tumor of a 13-year-old Japanese boy. ESFT is known to have a high rate of distant metastases at diagnosis. The genes controlling the spread of ESFT cells, however, have not been elucidated. G-banding chromosome analysis revealed that the MP-ASKIN-SA cell line has complex chromosomal abnormalities including trisomy 8. The EWS/FLI1 chimeric transcript and c-myc overexpression were revealed by the reverse transcriptase-polymerase chain reaction and Northern blot analysis. Furthermore, we investigated the expression of the focal adhesion kinase (FAK) gene in the ESFT cell lines using Northern blot analysis. In addition to the MP-ASKIN-SA cell line, six Ewing sarcoma cell lines, one peripheral nerve sheath tumor cell line, and two Askin tumor cell lines were analyzed. All ESFT cell lines, including MP ASKIN-SA, expressed five- to twenty-eight-fold-increased values of FAK, as compared with fibroblasts obtained from the bone marrow of a healthy volunteer. These results raise the possibility that the overexpression of c-myc and FAK are involved in the poor prognosis of ESFT. PMID- 14553944 TI - DNA gains at 8q23.2: a potential early marker in head and neck carcinomas. AB - Gains or amplifications involving chromosome arm 8q are one of the most recurrent chromosomal alterations in head and neck tumors. To characterize previously reported gains, we performed fluorescence in situ hybridization (FISH) using the sequences BAC RP1179E1 and 8-centromere PMJ 128 as probes. Gains and/or amplifications were detected in all 19 cases evaluated by FISH. The FISH analysis, but not G-banding, revealed homogeneously staining region in three cases. We conclude that gains of one or more genes on chromosome arm 8q may be important for the early stages of head and neck carcinomas. PMID- 14553945 TI - A new human cell line, PDSS-26, from poorly differentiated synovial sarcoma, with unique chromosomal anomalies. AB - Permanent synovial sarcoma cell lines are invaluable tools for understanding of the biology of this tumor. The present study reports the establishment of a new human cell line, PDSS-26, derived from a surgical specimen of a poorly differentiated synovial sarcoma. PDSS-26 has a doubling time of a 72 hours and grows as a monolayer of spindle cells that retain immunoreactivity for bcl-2 and vimentin. Karyotypic analysis revealed a rearrangement involving chromosomes 17 and 18, at the breakpoints q11.2 and q11.2, respectively, as the only structural aberrations. Analysis by reverse transcriptase polymerase chain reaction showed the presence of the SYT-SSX1 fusion transcript in both the primary tumor and the cell line. Cytoplasmic PTEN staining was detected by immunohistochemistry in both the PDSS-26 cell line and in original tumor, whereas no mutation was identified by automatic sequencing. Thus, PDSS-26 cells could be useful for future functional studies. PMID- 14553946 TI - Strong association between the GSTM1-null genotype and lung cancer in a Turkish population. AB - Glutathione S-transferases are possibly related to the detoxification of many xenobiotics involved in the etiology of cancer. To investigate the role of the glutathione S-transferase M1 deletion (GSTM1-null) in lung cancer, the polymerase chain reaction was used to determine the GSTM1 genotypes of lung cancer patients (n=101) and hospital (n=206) in a Turkish population. The prevalence of the GSTM1 null genotype in the case group was 48%, compared to 18% in the control group, giving an odds ratio (OR) of 4.14 (95% confidence interval [CI]=2.36-7.27). The analysis of patients by histologic type of lung cancer (10% adenocarcinoma, 43% squamous cell carcinoma, 26% small cell carcinoma, and 11% large cell carcinoma) showed no association between histopathologic type of lung cancer and GSTM1-null genotype. When the interaction between the GSTM1-null genotype and smoking status was analyzed, among the 67 smokers, the GSTM1-null genotype was found in 37 (55%) with an OR of 2.58 (95% CI=1.00-6.73) indicating a significant association. However, no association was found between smoking exposure (<30 and > or =30 packs/year) and GSTM1-null genotype. We conclude that, in this study the null GSTM1 genotype is an independent risk factor for the development of lung cancer for Turkish population. PMID- 14553947 TI - Differential deletions in 3p are associated with the development of head and neck squamous cell carcinoma in Indian patients. AB - In this study we performed detailed deletion mapping of two broad regions in the short arm (p) of chromosome 3 (i.e., 3p21.2 approximately p22 and 3p12 approximately p13), which were shown to have a high rate of deletions in head and neck lesions in our previous study. Using 18 highly polymorphic microsatellite markers, the deletion mapping was done in 35 dysplastic lesions and 46 primary head and neck squamous cell carcinoma (HNSCC) samples from Indian patients. Within the 21.6-megabase (Mb) region of 3p21.1 approximately p21.33, we have identified four areas (D1, 3p21.33; D2, 3p21.32; D3, 3p21.31; D4, 3p21.1) that showed a high frequency (46%-69%) of deletions in our samples. In the 3p12 approximately p13 region, we narrowed down the deletion within the 0.7-Mb region (D5, 3p12.1). Among these five regions (D1-D5), deletion in D3 is suggested to be necessary for the development of early dysplastic lesions, whereas the deletion in D2 may be necessary for dysplastic lesions and tumor progression. On the other hand, the deletion in D5 is significantly associated with progression of the lesions from mild/moderate to severe dysplasia. The deletions in D1 and D4, however, are required for tumor progression. As in our previous study, microsatellite size alterations (MA) were observed to be high in and around the highly deleted regions and gradually increased during the progression of the tumor. Loss of normal copy/interstitial alterations of chromosome 3 in the late stages of the tumor as well as rare biallelic alterations around the highly deleted regions also were seen in our samples. Human papilloma virus infection has been found to be associated with the deletion in the D5 region and MA in the D1 region, whereas nodal involvement of the tumor correlated only with the MA in D1 and D5. Thus, this study indicates that multiple tumor suppressor genes whose differential deletions are associated with the development of HNSCC may be present in 3p. PMID- 14553948 TI - Microsatellite alterations in hepatocellular carcinoma and intrahepatic cholangiocarcinoma. AB - A series of 20 hepatocellular carcinomas and 8 intrahepatic cholangiocarcinomas was screened from the Korean population for microsatellite alterations, including a loss of heterozygosity and replication errors using nine microsatellite markers containing several genes. The microsatellite results and our previous comparative genomic hybridization results of two tumors were compared at each locus, and the correlations between these and clinicopathologic variables were examined. The most characteristic findings were found at 13q. Replication errors were prevalent at D13S160 (13q21.2 approximately q31) and D13S292(13q12). The incidence of loss of heterozygosity, however, was higher at D13S153 (13q14.1 approximately q14.3) and D13S265(13q31 approximately q32). In contrast, there were higher deletion frequencies observed in hepatocellular carcinoma (HCC) and higher amplification frequencies observed in intrahepatic cholangiocarcinoma at 13q in our previous comparative genomic hybridization (CGH) study. Higher frequencies of replication errors were observed at D16S408 (13q12 approximately q21) and D16S504(13q23 approximately q24) in the HCC. This study found that significant differences in the patterns of genetic instability of microsatellites were dependent on the chromosomal loci. It is believed that certain genes at altered CGH regions, which are relevant to the development and/or progression of these cancers, are activated by different mutation mechanisms. PMID- 14553949 TI - Genetic and cytogenetic observations among different types of ovarian tumors are compatible with a progression model underlying ovarian tumorigenesis. AB - In this report we present the characterization of ovarian neoplasms including benign tumors, borderline tumors, and invasive carcinomas in order to assess whether a sharing of cytogenetic abnormalities is present in all three types of tumors. A cohort of 114 newly diagnosed and untreated ovarian epithelial tumors were analyzed by cytogenetic and molecular cytogenetic approaches with probes specific for chromosome 6. Three groups of chromosome abnormalities were identified: the first group included abnormalities common to all tumor classes (losses of chromosomes 6, 8, 10, 11, 15, 16, 17, 18, 19, 20, 21, 22, and X; gains of chromosomes 1, 3, 5, and 12; 6q24 approximately qter deletions); the second group presented specific abnormalities present in malignant but not in benign tumors (losses of chromosomes 2, 7, 13, and 14; gains of chromosome 4 and chromosome markers); and the last group included abnormalities unique to invasive carcinomas (loss of chromosome 4; gains of chromosomes 2, 7, 8, 9, 10, 16, 17, 18, 19, 20, and 21; 6q16 approximately q24 deletions; rearrangements of 3p, 3q, 13q, and 21q regions). The presence of shared chromosomal alterations in all three types of ovarian neoplasms investigated in this report seems therefore to suggest a progression model for these types of tumors. PMID- 14553950 TI - Interphase cytogenetic analysis in Argentinean B-cell chronic lymphocytic leukemia patients: association of trisomy 12 and del(13q14). AB - We have evaluated genomic aberrations by conventional cytogenetics and fluorescence in situ hybridization (FISH) analysis in a series of 57 Argentinean B-cell chronic lymphocytic leukemia (B-CLL) patients. The studies were performed on stimulated peripheral blood lymphocytes. FISH analysis for trisomy 12, 13q14 deletion, and monosomy of TP53 (also known as p53) was performed according to standard protocols. Our results showed 46.3% of patients with clonal chromosomal alterations by conventional cytogenetics and 80.7% by FISH. Trisomy 12 was found in 21.9% of patients by G-banding analysis and in 35% by FISH studies. Allelic loss of 13q14 was observed in 63.2% patients, most of them showing D13S319 and D13S25 deletion; 11% of patients showed TP53 monosomy. Coexistence of trisomy 12 and 13q14 deletion was found in 17.5% of patients. In this group, deletion 13q14 was the prevalent clone, with percentages 25-35% higher than those observed for trisomy 12, suggesting clonal evolution. The coexistence of trisomy 12 with deletion 13q14 was observed in a higher frequency than reported in the literature. A probable adverse prognosis is suggested for this group of patients, likely related to clonal evolution. PMID- 14553951 TI - Cytogenetic and comparative genomic hybridization findings in four cases of breast cancer after neoadjuvant chemotherapy. AB - To assess a potential common pattern of genetic alterations in chemotherapy resistant tumors we analyzed four tumors from breast cancer patients (patients 1 4) after neoadjuvant chemotherapy, by comparative genome hybridization (CGH) and conventional chromosome banding analysis. All patients showed structural aberrations involving chromosomes 1, 5, 11, 16, and 17. In CGH analysis, the patients showed typical imbalances for ductal breast cancer: gains of 1q (3 patients), 5q (2 patients), 8q (3 patients), and X (4 patients) and losses of 1p33 approximately p36 (3 patients), 16q (3 patients), 17p (3 patients), 19 (4 patients), and 22q (4 patients). Other recurrent imbalances of atypical pattern for ductal breast cancer were gain of 4q21 approximately q32 (2 patients), 20q21 approximately q22 (2 patients), and 21 (2 patients) and loss of 20p (3 patients). Three patients showed involvement of several regions bearing genes of drug resistance (MDR1 [HUGO symbol: ABCB1], BCRP [HUGO symbol: ABCG2], MRP1 [HUGO symbol: ABCC1], RFC1); the fourth patient displayed an amplification in the region of MYC (alias c-myc), thus providing--at the level of the light microscope -an explanatory background for the ability of their tumors to survive anthracycline-, taxane- and cyclophosphamide-based chemotherapy. Conventional cytogenetic analysis and CGH displayed highly coincidental findings in the tumors of four patients after neoadjuvant chemotherapy for breast cancer. PMID- 14553952 TI - ATM gene mutations are not involved in medulloblastoma in children. AB - Primitive neuroectodermal tumors (PNET)-medulloblastomas account for approximately 20% of all brain tumors in children. Ataxia-telangiectasia is an autosomal recessive neurological disorder with predisposition to cancer. The most common neoplasms are lymphoid malignancies and solid tumors, including central nervous system tumors, astrocytomas, and medulloblastomas. To investigate the potential role of the ATM gene in the pathogenesis of medulloblastoma, 13 tumors were screened for ATM mutations and 9 for loss of heterozygosity (LOH) of the ATM locus and flanking regions. In none of the tumors were mutations identified. In five of them, the well-known polymorphisms D1853N and F858L were identified and in all 22 tumors, the wild-type allele was preserved. The frequency of the polymorphisms was similar to that reported in our and other normal populations. The LOH of the 11q region (including the ATM gene), detected in 25% of informative cases, is consistent with the molecular and cytogenetic reports of deletion of chromosome 11 in 13%-41% of medulloblastomas. These results indicate that mutations in the ATM gene do not play a role in the pathogenesis of medulloblastoma in children. The LOH in the 11q region may suggest hidden unidentified tumor suppressor genes that may be involved in the malignant transformation. PMID- 14553953 TI - Cytogenetic analysis of the mononuclear cell component of giant cell tumors of bone. AB - This study comprises the cytogenetic analysis of mononuclear cells of a Mexican patient with a giant cell tumor of bone. This cell line showed nine translocations, two duplications, one addition, one deletion, and one ring chromosome, and did not present telomeric association. PMID- 14553954 TI - Centromeric transverse fission of chromosome 1 in a case of acute myelocytic leukemia. PMID- 14553955 TI - Severe megakaryocytic dysplasia in a case of myelodysplasia progressing to acute megakaryocytic leukemia presenting with dic(1;16)(q21;p13.3) and t(1;22)(p13;q13). PMID- 14553956 TI - A double-blind, placebo-controlled, dose-response study of the effectiveness and safety of lisinopril for children with hypertension. AB - BACKGROUND: Despite widespread use in hypertensive children, the safety and effectiveness of lisinopril had not been previously tested in a controlled study. METHODS: This study explored the dose-response relationship and safety of lisinopril in 115 hypertensive children, aged 6 to 16 years. Patients were randomized in a double-blind fashion for 2 weeks to one of three doses by body weight at baseline: <50 kg: low (0.625 mg), middle (2.5 mg), high (20 mg), and > or =50 kg: low (1.25 mg), middle (5 mg), high (40 mg). The dose-response for lisinopril was evaluated by analyzing the change in slope in sitting diastolic and systolic blood pressure (BP) by dose after 2 weeks of therapy compared to baseline. Patients then entered a double-blind withdrawal, where patients were either switched to placebo or continued their current lisinopril treatment for up to 2 weeks. Patients completed period II when their BP returned to baseline. Antihypertensive effectiveness, between placebo and lisinopril was determined for all doses. Adverse events were carefully monitored. RESULTS: There was a dose response relationship between the lowest and each of the higher doses of lisinopril. Blood pressure in the placebo group increased after withdrawal of lisinopril. The dose-response relationship was consistent across all subgroups (ie, age, Tanner stage, ethnicity, gender). CONCLUSIONS: Lisinopril, once daily, is an effective and well-tolerated antihypertensive in children aged 6 to 16 years. An initial dose of 0.07 mg/kg, administered once daily, effectively lowered BP within 2 weeks. Blood pressure was reduced in a dose-dependent fashion. PMID- 14553957 TI - Gestational calcium supplementation and blood pressure in the offspring. AB - BACKGROUND: The current study examined the relationship between calcium supplementation during pregnancy and blood pressure (BP) in the mother and offspring at 3 months and at 2 years postpartum. METHODS: Nulliparous pregnant women were assigned to either receive 2 g of calcium or placebo daily beginning between weeks 13 to 21 of gestation and continuing until delivery. Blood pressure was measured in children and their mothers at 3 months (n = 260) and (n = 57) at 2 years postpartum. Systolic BP was measured in the infants using a sphygmomanometer with ultrasonic amplification. For the toddlers, three supine BP measurements were taken from the right arm using a Critikon automated sphygmomanometer just after measurement of left ventricular wall thickness. RESULTS: Systolic BP in the calcium-supplemented infants was 2.2 mm Hg lower than in the placebo group (P >.05). At 2 years of age, systolic BP was 4.8 mm Hg lower in the calcium supplemented group (P <.05), whereas diastolic BP was 3 mm Hg lower (P >.05). There was no difference in left ventricular mass index between groups, although there was a significant correlation between systolic BP and wall thickness (P <.05). Maternal BP was positively correlated with circulating 1,25(OH)(2)D3 (P <.001) but did not differ between calcium groups at 3 months postpartum. CONCLUSIONS: The data on BP in the children are in agreement with previous studies and argue strongly for additional research into the effects of prenatal calcium supplementation on BP regulation in the offspring. PMID- 14553958 TI - Serum potassium and stroke risk among treated hypertensive adults. AB - BACKGROUND: In prospective studies, hypokalemia has been shown to be associated with a subsequent increase in stroke risk in treated hypertensive adults after 4 to 16 years of follow-up. Stroke risk associated with more recent assessments of hypokalemia has not been examined. METHODS: We used data from an on-going, population-based, case-control study of incident stroke at Group Health Cooperative (GHC). Cases were treated hypertensive adults, 30 to 79 years old, who sustained an incident ischemic or hemorrhagic stroke between July 1989 and December 2000. Controls were GHC members without a history of stroke who met same inclusion criteria as cases. Serum potassium (K(+)) levels were collected from GHC laboratory database. Hypokalemia (< or =3.4 mM/L) was defined using the most recent serum K(+) measure in the year before the index date (event date for cases; random date within calendar year of identification for controls). Multivariate logistic regression estimated the relative risk of ischemic and hemorrhagic stroke. RESULTS: Among 593 ischemic and 125 hemorrhagic stroke cases, and 2397 controls, few subjects were hypokalemic: 3%, 6%, and 2%, respectively. Using the normal range of serum K(+) as a reference (3.5 to 5.0 mM/L), hypokalemia was associated with an elevated risk of ischemic (odds ratio [OR]: 2.04; 95% confidence interval [CI]: 1.14-3.64) and hemorrhagic stroke (OR: 3.29; 95% CI: 1.45-7.48) in adjusted analyses. Associations were not modified by diuretic use. CONCLUSIONS: In adults with treated hypertension, hypokalemia in the year before a stroke was associated with an increased risk of incident ischemic and hemorrhagic stroke independent of diuretic use when compared to normal serum K(+) levels. PMID- 14553959 TI - Hemodynamic effects of chronic hemodialysis therapy assessed by pulse waveform analysis. AB - Cardiovascular responses to hemodialysis have been characterized by invasive monitoring techniques. These techniques are not feasible for evaluation of hemodynamic status during dialysis in the outpatient setting. In this study, we used pulse waveform analysis (PWA), a noninvasive tool designed for the ambulatory setting, to assess hemodynamic responses of dialysis treatments in 27 stable subjects with end-stage renal disease receiving chronic hemodialysis. In our population, systolic, diastolic, and pulse pressures were unaffected by dialysis despite the mean fluid removal of 3.0 +/- 0.2 kg. However, using PWA, we observed that stroke volume and cardiac output progressively declined by 17% to 19% (P <.001) with a concomitant increase in systemic vascular resistance by 22% from 1654 +/- 88 to 2020 +/- 121 dynes. sec. cm(-5) (P <.001). Also, we observed a significant reduction in small artery compliance from 4.7 +/- 0.5 to 3.3 +/- 0.4 mL. mm Hg(-1). 100 (P =.01), whereas large artery elasticity was unaffected. These findings suggest that changes in small artery vascular compliance contribute to the elevation in systemic vascular resistance during dialysis. This study confirms that hemodynamic adaptations to the dialysis procedure can be detected using PWA and are consistent with data obtained by invasive monitoring techniques. Furthermore, the observed reduction in vascular compliance in response to dialysis may contribute to the high cardiovascular risk in patients undergoing chronic hemodialysis therapy. PMID- 14553960 TI - Blood pressure and cognitive functioning among independent elderly. AB - BACKGROUND: The morbidity and mortality benefits of blood pressure (BP) control for the elderly is well documented. The cognitive consequences of hypertension control in this population, however, are still under debate. We aim to study the association between BP and cognitive performance in the elderly. Specifically, we explore 1) the possibility that BP is differentially associated with various cognitive domains; and 2) the utility of analyzing both BP and cognitive scores as continuous variables to unravel possible nonlinear associations. METHODS: Four hundred ninety-five community living 70 to 85 year olds completed eight cognitive tests that measured memory, concentration, visual retention, verbal fluency, and the mini-mental state examination (MMSE). The performance of each test was analyzed first by comparing four groups (normotensives, normalized hypertensives, untreated hypertensives, and treated but uncontrolled hypertensives). Then, using BP values as continuous variables, linear, U-curve, and J-curve associations were estimated. RESULTS: On all cognitive tests, except for verbal fluency, normotensives performed poorest, treated but uncontrolled hypertensives achieved the highest scores. The MMSE scores and the lighter concentration task were linearly related to BP; J-curve association was observed between memory and visual retention; prolonged concentration was related to pulse pressure alone. CONCLUSIONS: Low BP, as observed among the normotensive subjects, was associated with poor cognitive performance. Mild hypertension appeared to enhance cognitive functioning among the subjects of this study. Moreover, we found support for the hypothesis that the association between BP and different dimensions of cognition take on different patterns. PMID- 14553961 TI - Agonistic AT(1) receptor autoantibodies and monocyte stimulation in hypertensive patients. AB - BACKGROUND: Agonistic AT(1) receptor autoantibodies (AT(1)-AA) have been described in hypertensive and preeclamptic patients. Furthermore, monocytes are activated in hypertensive patients. We investigated and compared the ability of angiotensin II (Ang II) and AT(1)-AA to stimulate monocytes from hypertensive and normotensive persons. The adhesiveness of the monocytes to endothelial cell layers, tissue factor expression, and chemiluminescence were determined. METHODS: Blood samples were obtained from 17 patients with essential hypertension and from 20 normotensive subjects. Peripheral blood monocytes were isolated by Dynabeads and used in adhesion experiments. Adherence assays, Western blotting, and reactive oxygen species release by chemiluminescence were done. RESULTS: Monocyte adhesion to human aortic or umbilical vein endothelial cell layers was significantly higher after stimulation with AT(1)-AA, compared to Ang II or no stimulation. The effect was blocked with tissue factor antibody or epitope peptide preincubation. Eposartan was partially effective in blocking the effects. Western blotting after AT(1)-AA or Ang II stimulation showed that the monocytes expressed tissue factor. The AT(1)-AA and Ang II induced significantly higher chemiluminescence in monocytes from hypertensive than control subjects. Endothelial cells, on the other hand, showed much less chemiluminescence. CONCLUSIONS: These data show that monocytes can be stimulated by AT(1)-AA and Ang II to adhere, produce tissue factor, and probably reactive oxygen species. They underscore the importance of monocyte activation in hypertensive patients. The relevance of AT(1)-AA in hypertension will require further studies. PMID- 14553963 TI - beta-Adducin polymorphisms, blood pressure, and sodium excretion in three European populations. AB - The associations of the beta-adducin C1797T polymorphism with blood pressure (BP) and various indexes of sodium homeostasis were investigated in 388 men and 456 women, aged 18 to 60 years, recruited from three European populations (Cracow, Poland, n = 300; Novosibirsk, Russian Federation, n = 274; Mirano, Italy; n = 270). Phenotypes included 24-h ambulatory BP and urinary excretion of electrolytes and aldosterone. Subjects were genotyped for the beta-adducin polymorphism. Both a population-based association study and a family-based analysis were performed. Urinary sodium excretion was higher in Cracow than in Mirano (241 v 185 mmol/24 h, P <.05) and intermediate in Novosibirsk (206 mmol/24 h). The beta-adducin T allele (15.2% v 9.1%, P <.0001) was more prevalent in Mirano than in the two Slavic centers. In both population-based and family-based association analyses, there was significant heterogeneity between Slavic and Italian subjects in the phenotype-genotype relationships with beta-adducin. In the Slavic centers, 24-h systolic BP was higher in T allele carriers than in CC homozygotes (122.3 v 119.7 mm Hg, P =.03), whereas this was not the case in Mirano (121.8 v 122.9 mm Hg, P =.42). In Slavic (212.6 v 233.1 mmol/24 h) as well as in Italian (166.1 v 191.5 mmol/24 h) participants, 24-h sodium excretion was lower (P =.01) in T allele carriers than in CC homozygotes. These results were confirmed in the family-based analysis of offspring using a quantitative transmission disequilibrium test. In conclusion, the frequency of the beta adducin T allele and salt intake differ across European populations. Thus, both variation in genetic background and salt intake may explain the observed heterogeneity in the phenotype-genotype relationships. Genetic determinants of complex quantitative traits such as BP can only be investigated within their epidemiologic context. PMID- 14553962 TI - Effects of endothelial nitric oxide synthase, alpha-adducin, and other candidate gene polymorphisms on blood pressure response to hydrochlorothiazide. AB - BACKGROUND: Pharmacogenetic discoveries may enable greater individualization of antihypertensive drug therapy. We investigated polymorphisms in the genes encoding endothelial nitric oxide synthase (Glu298-->Asp), alpha-adducin (Gly460- >Trp), the beta(1)-adrenoceptor (Arg389-->Gly), beta2-adrenoceptor (Arg16-->Gly), and lipoprotein lipase (Ser447-->Stop) for their potential influences on blood pressure (BP) response to a thiazide diuretic. METHODS: The sample consisted of 291 unrelated non-Hispanic African American adults (150 women and 141 men) and 294 unrelated non-Hispanic white adults (126 women and 168 men) who were between 30 and 59.9 years of age and who had essential hypertension. Previous antihypertensive drug therapy was withdrawn for at least 4 weeks, and subjects were then treated with hydrochlorothiazide (25 mg daily) for 4 weeks to determine BP response. RESULTS: The covariates of ethnicity, gender, age, and waist-to-hip ratio accounted for 26% of interindividual variation in systolic BP response and 11% of interindividual variation in diastolic BP response. After adjustment for covariates, the endothelial nitric oxide synthase Glu298-->Asp polymorphism made an additional statistically significant contribution to predicting diastolic BP response to hydrochlorothiazide, accounting for another 1% of interindividual variation in response (P =.034). In contrast, the other polymorphisms, including the alpha-adducin Gly460-->Trp polymorphism, made no statistically significant contributions to prediction of BP response. CONCLUSIONS: Although we reject the null hypothesis of no genetic effects on BP response to hydrochlorothiazide, the influence of variation at single sites is likely to be small. More extensive characterization of genetic variation is required for pharmacogenetic approaches to become clinically useful in tailoring antihypertensive drug therapy for individual patients. PMID- 14553964 TI - T594M variant of the epithelial sodium channel beta-subunit gene and hypertension in individuals of African ancestry in South Africa. AB - BACKGROUND: The T594M variant of the beta-subunit of the sodium epithelial channel (ENaC) gene may contribute to hypertension in individuals of African origin. METHODS: A case-control study was performed to assess the role of the ENaC gene variant as an independent risk factor for hypertension in subjects of African ancestry. The effects of the ENaC gene variant on ambulatory blood pressure (BP) in hypertensive individuals and on office BP in hypertensive individuals and control subjects were also assessed. A total of 519 hypertensive patients with 24-h ambulatory BP (ABP) values determined while off medication, and 514 normotensive South African individuals of African ancestry were genotyped for the T594M polymorphism of the ENaC gene. RESULTS: A total of 22 (4.2%) hypertensive participants compared with 23 (4.5%) normotensive participants possessed the T594M variant (odds ratio = 1.06, confidence interval = 0.58 to 1.92, not significant). A similar genotype frequency distribution was noted in subjects representing the two predominant chiefdoms (Nguni and Sotho) in both case and control groups. No differences in frequency distribution of the T594M variant were noted with respect to either body mass index or gender. There were no differences in clinic or ambulatory mean, day, or night BP between hypertensive patients with or without the variant. Similarly, no differences were noted in clinic BP between control subjects with or without the variant. Other phenotypic parameters (including age and hypertension duration and severity) were also similar among hypertensive patients with or without the variant. CONCLUSION: These results do not support an important role for the T594M variant of the ENaC gene contributing to either the development or severity of hypertension in subjects of African ancestry. PMID- 14553965 TI - Disruption of the type 2 dopamine receptor gene causes a sodium-dependent increase in blood pressure in mice. AB - BACKGROUND: Dopamine D(2) receptors (D(2)Rs) are expressed in the kidney. It has not been determined whether D(2)Rs are involved in the mechanism of sodium handling and blood pressure (BP) control. METHODS: The function of D(2)Rs was investigated in mice disrupted with D(2)R gene (D(2)KO mice). Six-week-old male D(2)KO mice and wild-type (WT) mice were fed high-salt (4% NaCl) or low-salt (0.01% NaCl) diets for 8 weeks. RESULTS: Before starting the metabolic diet, there were no significant differences in body weight, food consumption, and 24-h urine excretions of creatinine, sodium and potassium. The high-salt diet caused a significant elevation in systolic BP in D(2)KO mice but not in WT mice. Calculation of sodium and potassium balances revealed a significantly high level of sodium retention in D(2)KO mice placed on the high-salt diet. Twenty-four-hour urine norepinephrine excretions and heart rates, indicators of sympathetic activity, were not different in D(2)KO and WT mice on the high-salt diet. Administration of nemonapride, a specific D(2)-like receptor antagonist, to WT mice given 0.9% NaCl in drinking water caused suppression of urinary sodium excretion but had no effect in mice without salt loading. CONCLUSIONS: These results suggest that D(2) receptors promote sodium excretion during a period of high salt intake. A defect in this mechanism may result in sodium-dependent BP elevation. PMID- 14553966 TI - Association of polymorphisms in the promoter region of the PNMT gene with essential hypertension in African Americans but not in whites. AB - BACKGROUND: Several studies have indicated that a region on human chromosome 17 may influence blood pressure. Our group reported positive linkage for hypertension to the region on human chromosome 17, between D17S1814 and D17S800 in white sibling pairs. In this study, we further investigated this result by examining the phenylethanolamine N-methyltransferase (PNMT) gene, which is located at 17q21 within the region where we found linkage. METHODS: A case/control association study was conducted to evaluate the relationship between genetic variants of the PNMT gene and risk for essential hypertension. Two single nucleotide polymorphisms (SNPs) in the promoter region of the gene were genotyped, PNMT-148 and PNMT-353, in three ethnic samples: African American (117 hypertensive, 96 normotensive), American white (91 hypertensive, 80 normotensive), and Greek white (99 hypertensive, 90 normotensive), using the homogeneous mass extend reaction (Sequenom) and RFLP for genotyping. RESULTS: A significant difference in allelic frequency of SNP-353 between hypertensives (38.02%) and normotensives (27.35%) in African Americans (P =.019) was found; however, no significant differences were observed for this SNP for the other ethnic groups. No association was found with SNP PNMT-148 in any of the ethnic groups. Frequencies of haplotypes based on the two SNPs were also compared between hypertensive and normotensive individuals. No significant difference was found in estimated haplotype frequencies between hypertensive and control subjects in the three ethnic groups. CONCLUSIONS: These results suggest that genetic variants of PNMT may play a role in the development of essential hypertension. PMID- 14553967 TI - Renal renin-angiotensin system activity in naturally reared and cross-fostered spontaneously hypertensive rats. AB - BACKGROUND: Young (4 week) spontaneously hypertensive rats (SHR) exhibit greater renal responses to angiotensin II (Ang II) than normotensive Wistar Kyoto (WKY) rats. SHR pups cross-fostering to a WKY dam at birth (SHRX) are less sensitive to Ang II and have lower adult blood pressure. The aim of this study was to compare renal renin-angiotensin system activity in young naturally reared and cross fostered SHR pups. METHODS: SHR and WKY rats were reared either by their natural mothers or by a foster mother of the opposite strain. At 5, 10, and 15 days of age, renal tissue renin activity and Ang II concentration were measured by radioimmunoassay. Renin-secreting cells were identified by in situ hybridization and AT(1) receptor expression was compared using Western blots. Ang II-mediated cAMP generation was measured in isolated proximal tubules. CONCLUSIONS: Tissue renin activity and numbers of renin-secreting cells did not differ, but Ang II was higher in SHRX. The AT(1) receptor expression was significantly lower in SHRX compared with SHR. Basal and Ang II-stimulated cAMP was lower in SHR tubules compared with WKY and SHRX tubules.Cross-fostering reversed the increased renal sensitivity of the SHR to Ang II. These data suggest that renal AT(1) receptor expression can be manipulated during the postnatal period and that this may affect adult blood pressure. PMID- 14553968 TI - beta-Blockers in hypertension-the emperor has no clothes: an open letter to present and prospective drafters of new guidelines for the treatment of hypertension. AB - Over the past decade, national and international guidelines have proposed beta blockers to be used on an equal footing with diuretics for initial therapy of hypertension. This preferred status was supposedly based on evidence documenting a reduction in morbidity and mortality with beta-blocker therapy in hypertension. We systematically analyzed all available outcome studies and found no evidence that beta-blocker based therapy, despite lowering blood pressure, reduced the risk of heart attacks or strokes. Despite the inefficacy of beta-blockers, the incidence of adverse effects is substantial. In the MRC study, for every heart attack or stroke prevented, three patients withdrew from atenolol because of impotence, and another seven withdrew because of fatigue. Thus the risk/benefit ratio of beta-blockers is characterized by lack of efficacy and multiple adverse effects. Given that many thorough, prospective, randomized trials attest to efficacy and safety of diuretics, calcium antagonists, ACE inhibitors, and angiotensin receptor inhibitors, the time has come to admit that beta-blockers should no longer be considered appropriate for first-line therapy in uncomplicated hypertension. PMID- 14553969 TI - Vascular effects of sildenafil in hypertensive cardiac transplant recipients. AB - BACKGROUND: Sildenafil is commonly used in the treatment of erectile dysfunction in hypertensive male cardiac transplant recipients (CTR); however, little is known about the vascular effects of sildenafil in these patients. METHODS: Central and peripheral arterial blood pressure (BP), heart rate, and brachial artery reactivity were determined in 15 hypertensive male CTR before and after oral sildenafil (50 mg) administration. RESULTS: Sildenafil improved brachial and aortic systolic BP, pulse pressure, aortic augmentation index, left ventricular tension time index, travel time of the reflected aortic pressure wave, and brachial artery reactivity (P <.01 for each comparison). No patient became hypotensive with sildenafil despite continuation of usual antihypertensive medications. CONCLUSIONS: Sildenafil (50 mg) is well tolerated in hypertensive CTR and improves BP, aortic augmentation index, and endothelial function. By decreasing the amplitude of the reflected pressure wave and delaying its return to the heart, sildenafil reduces left ventricular afterload and systolic stress. PMID- 14553970 TI - Blood pressure control in the hypertension clinic. AB - BACKGROUND: One effort to improve hypertension care in the United States is the development of the "Hypertension Specialist" program with appropriate training and certification of individual physicians. METHODS: We examined the impact of a hypertension clinic on blood pressure (BP) levels and control. RESULTS: By one year, BP declined 18/9 mm Hg, (P =.001) and BP control rates increased from 26% to 55% (P <.001) for systolic BP, from 47% to 82% (P <.001) for diastolic BP and from 18% to 52% (P <.001) for both. Age and systolic BP were significantly higher in uncontrolled patients. CONCLUSIONS: Attendance in a hypertension clinic significantly impacts BP control. PMID- 14553971 TI - Neurogenic essential hypertension revisited: the case for increased clinical and research attention. AB - The management of essential hypertension has increasingly focused on the use of diuretics, angiotensin-converting enzyme inhibitors, and angiotensin receptor blockers, which lower blood pressure (BP) through effects on blood volume and on the renin-angiotensin system. However, in many individuals these agents, whether given alone or in combination, fail to normalize BP. In such cases it is likely that hypertension is at least partly maintained by pathophysiologic mechanisms other than volume and the renin-angiotensin system, and therefore, that pharmacotherapy directed at other mechanisms is needed. One such form of hypertension is the often overlooked entity of neurogenic hypertension. The purpose of this article is to renew attention to this overlooked entity, to provide a very clinically oriented overview of its possible causes and manifestations, and to discuss the potentially important treatment implications of recognizing this form of hypertension. These implications underscore the need for further clinical and research attention concerning neurogenically mediated hypertension. PMID- 14553972 TI - Successes and shortcomings of the Food and Drug Modernization Act. PMID- 14553973 TI - Endothelial dysfunction in sustained and white coat hypertension. PMID- 14553974 TI - Regulatory mechanisms underlying decreased bone mineral density in women with essential hypertension. PMID- 14553976 TI - Synthesis of an octamannosyled glycan chain, the key oligosaccharide structure in ER-associated degradation. AB - The high-mannose type decasaccharide (Man(8)GlcNAc(2)), the proposed ligand of ER residing mannosidase-like proteins (MLP), and its monoglycosylated homologue (alpha-Glc(1)Man(8)GlcNAc(2)) were synthesized. The oligosaccharide assembly was performed in a convergent and stereoselective manner, using three oligosaccharide components, a core trisaccharide having a beta-mannoside bond, a liner mannotriose, and a branched mannotetraose. PMID- 14553977 TI - A practical synthesis of alpha-D-Manp-(1-->3)-alpha-D-Manp-(1-->2)-[alpha-D-Glcp (1-->3)]-alpha-D-Manp-(1-->2)-alpha-D-Manp-(1-->2)-alpha-D-Manp, an O-specific heterohexasaccharide fragment of Citrobacter braakii O7a, 3b, 1c. AB - An O-specific heterohexasaccharide fragment of Citrobacter braakii O7a, 3b, 1c, alpha-D-Manp-(1-->3)-alpha-D-Manp-(1-->2)-[alpha-D-Glcp-(1-->3)]-alpha-D-Manp-(1- >2)-alpha-D-Manp-(1-->2)-alpha-D-Manp was synthesized as its methyl glycoside. Acetylation of allyl 4,6-O-benzylidene-alpha-D-mannopyranoside, followed by debenzylidenization and benzoylation gave allyl 2,3-di-O-acetyl-4,6-di-O-benzoyl alpha-D-mannopyranoside (3), and subsequent deacetylation of 3 with CH(3)COCl MeOH gave the monosaccharide acceptor 4. Condensation of isopropyl 2,3,4,6-tetra O-benzyl-1-thio-beta-D-glucopyranoside (6) with 4 selectively afforded the alpha (1-->3)-linked disaccharide 7. Condensation of 7 with the (1-->3)-linked disaccharide donor 9, followed by deallylation and trichloroacetimidation, afforded the tetrasaccharide donor 12. Coupling of 12 with disaccharide acceptor 13, followed by debenzylation and deacylation, furnished the target heterohexasaccharide 16. PMID- 14553978 TI - Neighboring-group participation in benzylidene acetal ring-opening of a 2-cyano-2 deoxypyranoside derivative by diethylaluminum cyanide. AB - The oxirane ring-opening of an anhydro sugar with diethylaluminum cyanide (Et(2)AlCN) is a direct approach for obtaining a cyano derivative. Methyl 2,3 anhydro-4,6-O-benzylidene-alpha-D-allopyranoside showed anomalous chemical behavior when treated with Et(2)AlCN. The reaction afforded the corresponding beta-cyanohydrin as the minor component from a mixture of compounds resulting from the benzylidene acetal ring-opening caused by the attack of ethyl or cyano groups. PMID- 14553979 TI - Synthesis and characterisation of sulfated amphiphilic alpha-, beta- and gamma cyclodextrins: application to the complexation of acyclovir. AB - The synthesis of sulfated amphiphilic alpha-, beta- and gamma-cyclodextrins was achieved according to the standard protection-deprotection procedure. The formation of inclusion complexes between the amphiphilic alpha-, beta- and gamma cyclodextrins and an antiviral molecule, acyclovir (ACV) was investigated by UV visible spectroscopy (UV-Vis) and electrospray ionisation mass spectrometry (ESIMS). UV-Vis spectroscopy allowed determination of the stoichiometry and stability constants of complexes, whereas ESIMS, a soft ionisation technique, allowed the detection of the inclusion complexes. The results showed that the non sulfated amphiphilic cyclodextrins exhibit a 1:2 stoichiometry with acyclovir, while sulfated amphiphilic cyclodextrins, except gamma-cyclodextrin, exhibit a 1:1 stoichiometry indicating the loss of one interaction site. Non-covalent interactions between acyclovir and non-sulfated amphiphilic cyclodextrins appear to take place both in the cavity of the cyclodextrin and inside the hydrophobic zone generated by alkanoyl chains. In contrast, in the case of sulfated amphiphilic cyclodextrins, the interactions appear to involve only the hydrophobic region of the alkanoyl chains. PMID- 14553980 TI - FTIR study of state and phase transitions of low moisture sucrose and lactose. AB - Mid-infrared spectra of freeze-dried sucrose and lactose systems were acquired over a range of temperatures (30-200 degrees C) and water contents (0-6.3%). Starting from the glassy state, the experimental conditions were selected to cover the main thermal transitions: the glass-rubber transition, the crystallisation and, for some samples, the subsequent melting. The FTIR spectra were very sensitive to the physical state. While subtle but systematic spectral differences between the glassy and rubbery states were detectable throughout the spectrum, a very pronounced increase in spectral resolution was observed as crystallisation occurred and was followed by the expected spectral broadening during melting. The temperatures at which these changes occurred were in satisfactory agreement with the transition temperatures measured by differential scanning calorimetry (DSC). The increase in molecular mobility as a result of increasing temperature or plasticisation by water led to a significant shift of the O-H stretching band to higher wavenumbers indicating a weakening of hydrogen bonding. This shift reached a maximum as the DSC measured crystallisation temperature range was approached. As expected, the crystallisation led to a highly effective hydrogen bonding network. This was more significant for lactose than for sucrose. No significant step change in hydrogen bonding was observed at Tg. As anticipated, the temperature at which these transitions occurred decreased with increasing water content but overlapped when observed in the context of the shifted temperature (T-Tg). PMID- 14553981 TI - Synthesis of beta-(1-->6)-branched (1-->3)-glucododecaose and -glucopentadecaose with alternate beta- and alpha-bonds in the backbone. AB - Beta-D-Glcp-(1-->3)-[beta-D-Glcp-(1-->6)]-alpha-D-Glcp-(1-->3)-[beta-D-Glcp-(1- >3)-[beta-D-Glcp-(1-->6)]-alpha-D-Glcp-(1-->3)](2-3)-beta-D-Glcp-(1-->3)-[beta-D Glcp-(1-->6)]-beta-D-Glcp were synthesized as their methoxyphenyl glycosides in a concise way with a trisaccharide as the building block. PMID- 14553982 TI - A synergistic reaction mechanism of a cycloalternan-forming enzyme and a D glucosyltransferase for the production of cycloalternan in Bacillus sp. NRRL B 21195. AB - Cycloalternan-forming enzyme (CAFE) was first described as the enzyme that produced cycloalternan from alternan. In this study, we found that a partially purified preparation of CAFE containing two proteins catalyzed the synthesis of cycloalternan from maltooligosaccharides, whereas the purified CAFE alone was unable to do so. In addition to the 117 kDa CAFE itself, the mixture also contained a 140 kDa protein. The latter was found to be a disproportionating enzyme (DE) that catalyzes transfer of a D-glucopyranosyl residue from the non reducing end of one maltooligosaccharide to the non-reducing end of another, forming an isomaltosyl residue at the non-reducing end. CAFE then transfers the isomaltosyl residue to the non-reducing end of another isomaltosyl maltooligosaccharide, to form an alpha-isomaltosyl-(1-->3)-alpha-isomaltosyl-(1- >4)-maltooligosaccharide, and subsequently catalyzes a cyclization to produce cycloalternan. Thus, DE and CAFE act synergistically to produce cycloalternan directly from maltodextrin or starch. PMID- 14553984 TI - Imazalil-cyclomaltoheptaose (beta-cyclodextrin) inclusion complex: preparation by supercritical carbon dioxide and 13C CPMAS and 1H NMR characterization. AB - An inclusion complex between imazalil (IMZ), a selected fungicide, and cyclomaltoheptaose (beta-cyclodextrin, betaCD) was obtained using supercritical fluid carbon dioxide. The best preparation conditions were determined, and the inclusion complex was investigated by means of 1H NMR spectroscopy in aqueous solution and 13C CPMAS NMR spectroscopy in the solid state. Information on the geometry of the betaCD/IMZ complex was obtained from ROESY spectroscopy, while the dynamics of the inclusion complex in the kilohertz range was obtained from the proton spin-lattice relaxation times in the rotating frame, T(1rho) (1H). PMID- 14553983 TI - Preparation and reactivity of a novel disaccharide, glucosyl 1,5-anhydro-D fructose (1,5-anhydro-3-O-alpha-glucopyranosyl-D-fructose). AB - A novel disaccharide, glucosyl 1,5-anhydro-D-fructose (1,5-anhydro-3-O-alpha glucopyranosyl-D-fructose, GAF) was enzymatically prepared from 1,5-anhydro-D fructose (1,5-AF) and cyclomaltoheptaose (beta-cyclodextrin). Cyclodextrin glucanotransferase transferred various sizes of maltooligosaccharide to 1,5-AF. Glucoamylase digested the maltooligosyl chain of the products to a glucosyl residue giving a final product, GAF. An NMR analysis of GAF elucidated that the glucose residue was linked to C-3 of the 1,5-AF residue with an ether linkage. Reactivity on the aminocarbonyl reaction of GAF with bovine serum albumin was lower than that of 1,5-AF, but was higher than that of glucose. PMID- 14553985 TI - Forster energy-transfer studies between Trp residues of alpha1-acid glycoprotein (orosomucoid) and the glycosylation site of the protein. AB - Energy-transfer studies between Trp residues of alpha(1)-acid glycoprotein and the fluorescent probe Calcofluor White were performed. Calcofluor White interacts with carbohydrate residues of the protein, while the three Trp residues are located at the surface (Trp-160) and in hydrophobic domains of the protein (Trp 25 and Trp-122). Binding of Calcofluor to the protein induces a decrease in the fluorescence intensity of the Trp residues accompanied by an increase of that of Calcofluor White. Efficiency (E) of Trp fluorescence quenching was determined to be equal to 45%, and the Forster distance R(o), at which the efficiency of energy transfer is 50%, was calculated to be 18.13 A. This low distance and the value of the efficiency clearly indicate that energy transfer between Trp residues and Calcofluor White is weak. PMID- 14553986 TI - A mild and convenient indium(III) chloride-catalyzed synthesis of thioglycosides. AB - The efficiency of glycosidation reactions generally involves a high chemical yield, as well as high/complete stereo- and regioselectivity. All these depend on the compatibility of the reactivity of glycosyl donors and acceptors. Among glycosyl donors, thioglycosides are widely used because of their high degree of stability in many organic reactions. Although there are number of methods available for the preparation of thioglycosides, all of them have one or more disadvantages, especially concerning the time factor and cumbersome workup procedures. Here we report a convenient and high-yielding method for the preparation of thioglycosides. PMID- 14553987 TI - Efficient synthesis of 5-thio-D-arabinopyranose and 5-thio-D-xylopyranose from the corresponding D-pentono-1,4-lactones. AB - 5-Thio-D-arabinopyranose (5) and 5-thio-D-xylopyranose (10) were synthesized from the corresponding D-pentono-1,4-lactones. After regioselective bromination at C 5, transformation into 5-S-acetyl-5-thio derivatives, reduction into lactols and deprotection afforded the title compounds in 49 and 42% overall yield, respectively. PMID- 14553988 TI - Alkaline chitosan solutions. AB - Rigid and transparent hydrogels were obtained upon pouring chitosan salt solutions into saturated ammonium hydrogen carbonate. Incubation at 20 degrees C for 5 days yielded chitosan carbamate ammonium salt, Chit-NHCO(2)(-)NH(4)(+) a chemical species that either by hydrolysis or by thermal treatment decomposed to restore chitosan in free amine form. Chitosans of different degrees of acetylation, molecular sizes and origins (squid and crustaceans) were used as hydrochloride, acetate, glycolate, citrate and lactate salts. Their hydrogels obtained in ammonium hydrogen carbonate yielded chitosan solutions at pH values as high as 9.6, from which microspheres of regenerated chitosans were obtained upon spray-drying. These materials had a modest degree of crystallinity depending on the partial acylation that took place at the sprayer temperature (168 degrees C). Citrate could cross-link chitosan and impart insolubility to the microspheres. Chloride on the contrary permitted to prepare microspheres of chitosan in free amine form. By the NH(4)HCO(3) treatment, the cationicity of chitosan could be reversibly masked in view of mixing chitosan with alginate in equimolar ratio without coacervation. The clear and poorly viscous solutions of mixed chitosan carbamate and alginate were spray-dried at 115 degrees C to manufacture chitosan-alginate microspheres having prevailing diameter approx 2 micron. PMID- 14553989 TI - Global pollution monitoring of butyltin compounds using skipjack tuna as a bioindicator. AB - Butyltin compounds (BTs) including mono- (MBT), di- (DBT), tri-butyltin (TBT) and total tin (sigmaSn), were determined in the liver of skipjack tuna (Katsuwonus pelamis) collected from Asian offshore waters (off-Japan, the Japan Sea, off Taiwan, the East China Sea, the South China Sea, off-Philippines, off-Indonesia, the Bay of Bengal), off-Seychelles, off-Brazil and open seas (the North Pacific). BTs were detected in all the skipjack tuna collected, suggesting widespread contamination of BTs even in offshore waters and open seas on a global scale. Considering specific accumulation, Sex-, body length- differences and migration of skipjack tuna did not seem to affect BT concentrations, indicating rapid reflection of the pollution levels in seawater where and when they were collected. Skipjack tuna is a suitable bioindicator for monitoring the global distribution of BTs in offshore waters and open seas. High concentrations of BTs were observed in skipjack tuna from offshore waters around Japan, a highly developed and industrialized region (up to 400 ng/g wet weight). Moreover skipjack tuna collected from offshore waters around Asian developing countries also revealed the levels comparable to those in Japan (up to 270 ng/g wet weight) which may be due to the recent improvement in economic status in Asian developing countries. High percentages (almost 90%) of BTs in total tin (sigmaSn: sum of inorganic tin+organic tin) were found in the liver of skipjack tuna from offshore waters around Asian developing countries. This finding suggests that the anthropogenic BTs represent the major source of Sn accumulation in skipjack tuna from these regions. PMID- 14553990 TI - Mutual effects of cadmium and phosphate on their adsorption and desorption by goethite. AB - Adsorption of cadmium (Cd) and phosphate by oxides or soils has been extensively studied, but the adsorption/desorption kinetics and mutual effects of these two species in co-existing systems has received little attention. In this study, a batch equilibration method was used to investigate the effect of phosphate and its application time on Cd adsorption and desorption on goethite. The influence of Cd and its application time on phosphate sorption and desorption kinetics was also determined. For Cd adsorption, phosphate was introduced into the system by two sequences: pre-treating goethite at 40 (degrees)C for 1 week, and applying with Cd simultaneously. Similarly, for phosphate sorption, Cd was applied by pre treating goethite at 40 (degrees)C for 1 week or simultaneous addition with phosphate. Results demonstrated that phosphate added to goethite enhanced Cd adsorption, and facilitated Cd release as compared to untreated goethite. Cadmium had slightly higher adsorption, but a significantly faster desorption rate from the goethite simultaneously treated with phosphate and Cd, as compared to phosphate-pretreated goethite. Cadmium and its application time had little impact on phosphate sorption by goethite. However, phosphate desorption kinetics was affected by Cd application time. When the sorption time was short (15 min), phosphate desorption was faster from the goethite that was simultaneously treated with phosphate and Cd, as compared to Cd pretreated or untreated goethite. In contrast, a longer sorption time (4 weeks) resulted in a higher desorption rate of phosphate from Cd pretreated goethite than simultaneously phosphate-Cd treated goethite. This study provided useful information on adsorption/desorption kinetics in complicated Cd-phosphate-goethite systems. PMID- 14553991 TI - Soil ecotoxicity assessment using cadmium sensitive plants. AB - Four crop plant species (sweet corn, Zea may; wheat, Triticum aestivum; cucumber, Cucumis sativus; and sorghum, Sorghum bicolor) were tested to assess an ecotoxicity in cadmium-amended soils. The measurement endpoints used were seed germination and seedling growth (shoot and root). The presence of cadmium decreased the seedling growth. The medium effective concentration values (EC50) for shoot or root growth were calculated by the Trimmed Spearman-Karber method. Due to the greater accumulation of Cd to the roots, root growth was a more sensitive endpoint than shoot growth. Bioavailability and transport of Cd within plant were related to concentration and species. The ratio of bioaccumulation factor (BAF) in the shoots to the roots indicated high immobilization of Cd in the roots. Seed germination was insensitive to Cd toxicity, and is not recommended for a suitable assay. Among the test plants and test endpoints, root growth of sorghum and cucumber appears to be a good protocol to assess ecotoxicity of soils contaminated by Cd. PMID- 14553992 TI - Foraminiferal proxies for pollution monitoring in moderately polluted harbors. AB - Benthic foraminifera are increasingly used as environmental bio-indicators, especially in polluted environments where their sensitivity to pollutants may be expressed by a modification of the assemblages. Eighteen sediment samples were collected in September 2000 in five harbors located in moderately polluted estuaries on the coast of Vendee (France) for the study of foraminiferal assemblages. Ten heavy metals and 13 PAH have been analyzed from the sediments. The marine to continental estuarine gradient has a prevalent influence on the foraminiferal distribution. However, the results show that foraminiferal density and species richness of the assemblages decrease with an increase in heavy metal and PAH concentration, and therefore may be used as pollution indicators. Moreover, the more polluted areas are dominated by the tolerant pioneer species Haynesina germanica that may be used as bio-indicator of pollution, mainly in the uppermost areas. PMID- 14553993 TI - Ability of bacterial biphenyl dioxygenases from Burkholderia sp. LB400 and Comamonas testosteroni B-356 to catalyse oxygenation of ortho hydroxychlorobiphenyls formed from PCBs by plants. AB - Capacity of enzymes of the biphenyl/chlorobiphenyl pathway, especially biphenyl dioxygenase (BPDO) of two polychlorinated biphenyls (PCB) degrading bacteria, Burkholderia sp. LB400 and Comamonas testosteroni B-356, to metabolize ortho substituted hydroxybiphenyls was tested.,These compounds found among plant products of PCB metabolism, are carrying chlorine atoms on the hydroxyl substituted ring. The abilities of His-tagged purified LB400 and B-356 BPDOs to catalyze the oxygenation of 2-hydroxy-3-chlorobiphenyl, 2-hydroxy-5 chlorobiphenyl and 2-hydroxy-3,5-dichlorobiphenyl were compared. Both enzyme preparations catalyzed the hydroxylation of the three chloro-hydroxybiphenyls on the non-substituted ring. Neither LB400 BPDO nor B-356 BPDO oxygenated the substituted ring of the ortho-hydroxylated biphenyl. The fact that metabolites generated by both enzymes were identical for all three hydroxychlorobiphenyls tested; exclude any other mode of attack of these compounds by LB400 BPDOs than the ortho-meta oxygenation. PMID- 14553994 TI - Preliminary study of prairies forested with Eucalyptus sp. at the northwestern Uruguayan soils. AB - The land cover change of Uruguayan Forestal Plan provoked biogeochemical changes on horizon Au(1) of Argiudols; in native prairies which were replaced by monoculture Eucalyptus sp. plantation with 20 year rotations as trees. Five fields forested and six natural prairies were compared. The results not only show a statistical significant soil acidification, diminution of soil organic carbon, increase of aliphaticity degree of humic substances, and increase of affinity and capacity of hydrolytic activity from soil microbial communities for forested sites with Eucalyptus sp. but also, a tendency of podzolization and/or mineralization by this kind of land cover changes, with a net soil organic lost of 16.6 tons ha(-1) in the horizon Au(1) of soil under Eucalyptus sp. plantation compared with prairie. Besides, these results point out the necessity of correction of the methodology used by assigned Uruguayan commission to assess the national net emission of greenhouse gases, since the mineralization and/or podzolization process detected in forested soil imply a overestimation of soil organic carbon. The biochemical parameters show a statistical significant correlation between the soil organic carbon status and these parameters which were presented as essential for the correct evaluation of Uruguayan soil carbon sink. PMID- 14553995 TI - Throughfall chemistry and canopy interactions in a Sitka spruce plantation sprayed with six different simulated polluted mist treatments. AB - Throughfall chemistry was studied in a mature Sitka spruce plantation in order to investigate canopy interactions, such as nitrogen absorption, cation leaching, and neutralization of rainfall passing through the canopy. The plantation had been exposed to six different simulated mist treatments including N (NH(4)NO(3)) and S (H(2)SO(4) at pH 2.5) in four replicated blocks since 1996. Throughfall and rainfall were collected from May to September 2000. The results showed that 30 35% of the applied N was retained by the canopy. There were linear relationships between the loss of H(+) and increased K(+), Mg(2+) and Ca(2+) deposition through the canopy. However these increases in K(+), Mg(2+) and Ca(2+) deposition accounted for only about 50% of total neutralization of the acidity. The relationship between the anion deficits in throughfall and the loss of H(+) implied that weak organic acid anions were involved in the neutralization of the acidity in throughfall. PMID- 14553996 TI - White birch (Betula papyrifera Marshall) foliar litter decomposition in relation to trace metal atmospheric inputs at metal-contaminated and uncontaminated sites near Sudbury, Ontario and Rouyn-Noranda, Quebec, Canada. AB - Decomposition of white birch (Betula papyrifera Marshall) foliar litter was examined at metal-contaminated and uncontaminated sites established along gradients of soil Cu, Ni, Pb and Zn concentrations near Sudbury, Ontario and Rouyn-Noranda, Quebec. Over an 18-month study period, a significantly lower rate of litter mass loss was observed at the Sudbury contaminated site (S1) than at the uncontaminated site (S2). This result was not duplicated at corresponding sites (RN1, RN2) in Rouyn-Noranda, despite similar levels of soil metal contaminants and atmospheric inputs. Concentrations of metals in litter increased at all sites with time. However, the greatest litter Cu and Ni concentrations were observed at S1 (188 and 192 microg/g, respectively), a result of substantial net gains of these elements from atmospheric inputs. On a per hectare basis, Cu accumulation in litter at S1 approached recommended application rates of Cu as copper sulphate for control of fungal diseases in agricultural operations, indicating that the current rate of Cu smelter emissions in Sudbury may cause the observed impairment of decomposition. PMID- 14553997 TI - Evaluation of chemical immobilization treatments for reducing heavy metal transport in a smelter-contaminated soil. AB - Three chemical immobilization materials, agricultural limestone (AL), mineral rock phosphate (RP), and diammonium phosphate (DAP), were evaluated using solute transport experiments to determine their ability to reduce subsurface heavy metal transport in a smelter contaminated soil. Percent reductions in metals transported were based on comparison with cumulative totals of metal species eluted through 60 pore volumes from an untreated soil. Reductions of metal eluted from the AL treatment were 55% for Cd, 45.2% for Pb, and 21.9% for Zn. Rock phosphate mixed with soil at 60 and 180 g kg(-1) was generally ineffective for reducing Cd, Pb, and Zn elution with <27% reduction for Cd, Pb, and Zn. Rock phosphate placed under contaminated soil as a reactive barrier (i.e. layered RP) at 180 g kg(-1) reduced Cd 53% and Zn 24%, and was the most efficient treatment for reducing Pb (99.9%) transport. DAP treatments were superior to all other materials for reducing Cd and Zn elution with reduction >77% for Zn and >91% for Cd from the 90 g DAP kg(-1) treatment. Increasing DAP from 10 to 90 g kg(-1) increased total arsenic released from 0.13 to 29.5 mg kg(-1) and total P eluted from 2.31 to 335 mg. DAP at 10 g kg(-1) was the most effective treatment for immobilizing the combination of Cd, Pb, and Zn, with reductions of 94.6, 98.9, and 95.8%, respectively. PMID- 14553998 TI - Comparison of trace element accumulation in Baikal seals (Pusa sibirica), Caspian seals (Pusa caspica) and northern fur seals (Callorhinus ursinus). AB - Concentrations of 18 trace elements (V, Cr, Mn, Co, Cu, Zn, Se, Rb, Sr, Zr, Mo, Ag, Cd, Sb, Cs, Hg, Tl and Pb) were determined in liver, kidney, muscle and hair of Baikal seals, Caspian seals and northern fur seals. All the three species showed the highest concentrations of Hg, V, Mn, Se and Ag in liver, Cd, Co and Tl in kidney, and Cs in muscle among the soft tissues examined. The highest burdens of Zn, Rb and Cs were observed in muscle, Mo and Ag in liver, and Sb and Pb in hair in all the three species. Concentrations of non-essential elements, Rb, Cd, Cs and Hg, showed significant positive correlations among liver, kidney and muscle, whereas correlation coefficients for essential elements, Mn, Co, Cu, Zn and Se, between the three tissues were generally low for all the species, suggesting that homeostasis controls the concentrations of essential elements but not the non-essential elements in the tissues of these animals. Significant age dependent increase was found in the concentrations of V, Se and Ag in liver and Hg in liver and kidney of all the three species. Hair concentrations showed significant positive correlations with Zn levels in liver and kidney and Hg in muscle for Caspian seals, Hg in liver and kidney for Baikal seals, and Pb in liver for northern fur seals. Furthermore, regression analysis using the data in the present study and in the literature showed significant positive correlations between Hg levels in hair, and liver, kidney and muscle for various species of pinnipeds. These results indicate the possibility of using hair samples for monitoring these trace elements in pinnipeds. PMID- 14553999 TI - Combined effects of copper and food on the midge Chironomus riparius in whole sediment bioassays. AB - Effects observed in whole-sediment bioassays must be seen as the joint effect of all sediment characteristics. In whole-sediment bioassays, however, adverse effects on test organisms are usually attributed to the presence of contaminants and effects of food are often ignored. The aim of this study was to analyze the response of the midge Chironomus riparius to sediment spiked with different combinations of food and copper. The responses of C. riparius to these spiked sediments were assessed in 10-day whole-sediment bioassays. Decreases in survival, dry weight, and length of C. riparius were observed with increasing copper concentrations. However, an increase in the amount of food resulted in an increase of larval dry weight and length until copper concentrations reached a critical threshold of 200 mg/kg. In addition, an increase in the amount of food resulted in a decrease of accumulated copper in the larvae. The present study demonstrated that the combination of copper and food in the sediment determines the performance of C. riparius in whole-sediment bioassays. The dependency of C. riparius on high feeding levels, which mask toxic effects, questions its suitability as a test organism for whole-sediment bioassays. Because benthic communities in polluted ecosystems are often exposed to varying levels of both food and toxicants it is concluded that the trophic state of the ecosystem may alter the ecological risk of sediment-bound toxicants to opportunistic benthic invertebrates such as C. riparius. PMID- 14554000 TI - Regression models to predict water-soil heavy metals partition coefficients in risk assessment studies. AB - Risk assessment studies apply fate and transport models to predict the behaviour of chemicals in the environment. The definition of physico-chemical properties is crucial to predict the mobility of pollutants and heavy metals in particular within the environmental compartments. The conservative approach normally adopted at a screening level in attributing a value to the K(d) value, results in an extremely variable mobility in soil. In this paper a regression model to estimate rapidly the K(d) for heavy metals is proposed and applied to Pb, allowing a considerable reduction (3-4 orders of magnitude) of the estimation uncertainty. The application of a stepwise forward multiple regression to literature data provided a pH-dependent regression equation of the soil-water distribution coefficient (K(d)) for Pb: log K(d)=1.99+0.42 pH. PMID- 14554001 TI - Cadmium accumulation and elimination in tissues of juvenile olive flounder, Paralichthys olivaceus after sub-chronic cadmium exposure. AB - Experiments were carried out to investigate the accumulation and elimination of cadmium (Cd) in tissues (gill, intestine, kidney, liver and muscle) of juvenile olive flounder, Paralichthys olivaceus, exposed to sub-chronic concentrations (0, 10, 50, 100 microg l(-1)) of Cd. Cd exposure resulted in an increased Cd accumulation in tissues of flounder with exposure periods and concentration, and Cd accumulation in gill and liver increased linearly with the exposure time. At 20 days of Cd exposure, the order of Cd accumulation in organs was gill > intestine > liver > kidney > muscle and after 30 days of exposure, those were intestine > gill > liver > kidney > muscle. An inverse relationship was observed between the accumulation factor (AF) and the exposure level, but AF showed an increase with exposure time. During the depuration periods, Cd concentration in the gill, intestine and liver decreased immediately following the end of the exposure periods. No significant difference was found Cd in concentration in the kidney and muscle during depuration periods. The order of Cd elimination rate in organs were decreased intestine > liver > gill during depuration periods. PMID- 14554002 TI - High human exposure to lead through consumption of birds hunted with lead shot. AB - We assess lead contamination of Greenland seabirds killed with lead shot having studied thick-billed murre and common eider, the two most important species in the diet. The lead concentration is very high in meat of eiders killed with lead shot (mean 6.1 microg/g-wet wt, 95% CL 2.1-12). This level is about 44 times higher than in drowned eiders and eight times higher than in shot murres. Analyzing whole breasts instead of sub-samples reveals about seven times higher lead levels in birds' meat. We conclude that in some cases the lead intake by Greenland bird eaters will largely exceed the FAO/WHO tolerable lead intake guideline and that lead shot is a more important source of lead in the diet than previously estimated. PMID- 14554003 TI - Characterisation of a reference site for quantifying uncertainties related to soil sampling. AB - The paper reports a methodology adopted to face problems related to quality assurance in soil sampling. The SOILSAMP project, funded by the Environmental Protection Agency of Italy (APAT), is aimed at (i) establishing protocols for soil sampling in different environments; (ii) assessing uncertainties associated with different soil sampling methods in order to select the "fit-for-purpose" method; (iii) qualifying, in term of trace elements spatial variability, a reference site for national and international inter-comparison exercises. Preliminary results and considerations are illustrated. PMID- 14554004 TI - Soil mobility of sewage sludge-derived dissolved organic matter, copper, nickel and zinc. AB - A soil (sandy loam) column leaching study aimed to determine the extent of mobility and co-mobility of Cu, Ni, Zn and dissolved organic matter (DOM) released from a surface-application (equivalent to 50 t ds ha(-1) of anaerobically-digested sewage sludge. Leaching of DOM through the soil column was found to be almost un-retarded. Decidedly similar behaviour was exhibited by Ni suggesting that it migrated as organic complexes. Whilst Cu was also found to be leached, significant retardation was evident. However, the importance of DOM in promoting the mobility of both Cu and Ni was evidenced by their lack of mobility when added to the soil column as inorganic forms. The presence of DOM did not prevent Zn from becoming completely adsorbed by the soil solid phase. In relation to WHO drinking water guidelines, only Ni concentrations showed potential environmental significance, due to the relatively poor retention of Ni by the sludge solid phase. PMID- 14554005 TI - Genotype-specific responses of fluctuating asymmetry and of preadult survival to the effects of lead and temperature stress in Drosophila melanogaster. AB - Although fluctuating asymmetry (FA) increases with exposure to certain types of environmental stressors such as temperature extremes, relatively little is known about the effects of interaction (e.g., synergism) between known sources of environmental stress on FA. Knowledge of such interaction effects, and of the magnitude of genotype-by-environment interaction, are of fundamental importance toward predicting the usefulness of FA as a bioindicator of environmental pollution. We tested for synergistic effects on FA between elevated temperature and exposure to lead, and examined FA responses simultaneously in four genetic strains of Drosophila melanogaster known to differ in their degree of developmental instability, and presumably in their buffering capacity. In the absence of heavy metal, bristle FA increased with temperature, but in the presence of lead, FA at high temperature (30 (degrees)C) was reduced to levels similar, or below, that at lower temperature (25 (degrees)C). This temperature by lead interaction was statistically significant, but paradoxical in that the disruptive effects of temperature appeared to be attenuated in the presence of the heavy metal. In no case was there a significant effect of lead on bristle FAs, despite documented assimilation of heavy metal by flies, and in no case was the genotype by environment interaction significant. Whereas lead treatment did not influence survival, survival was reduced at the high temperature, but significantly so only in one genetic strain (Oregon-R). There was no relationship between survival and FA across stress treatments within lines. Thus, any disproportionate stress-induced mortality in developmentally unstable classes (developmental selection) was unlikely to bias the FA results. Our results underscore the need for independent replication of significant findings before FA based biomonitoring can be responsibly and effectively implemented. The results call for caution in using FA as a biomarker of stress, because stress factors may interact in complex and unpredictable ways, which could result in erroneous conclusions about real levels of stress present in field populations, under the unduly simplistic assumption that stress factors will act additively to increase FA. PMID- 14554006 TI - Neuregulin: a cut-and-pasted cancer gene? PMID- 14554007 TI - Promoting proteasomes: trash to treasure. PMID- 14554008 TI - Pharmacists seek the solution of a shaman. PMID- 14554011 TI - Mark Chandler discusses rules-based medicine and multi-analyte profiling. Interview by Stephen L Carney. PMID- 14554012 TI - A 'rule of three' for fragment-based lead discovery? PMID- 14554013 TI - PET and knockout mice in drug discovery. PMID- 14554015 TI - Multidimensional proteomics. PMID- 14554016 TI - Prevention and treatment of bacterial diseases caused by bacterial bioterrorism threat agents. AB - There is general consensus that the bacterial agents or products most likely to be used as weapons of mass destruction are Bacillus anthracis, Yersinia pestis, Francisella tularensis and the neurotoxin of Clostridium botulinum. Modern supportive and antimicrobial therapy for inhalational anthrax is associated with a 45% mortality rate, reinforcing the need for better adjunctive therapy and prevention strategies. Pneumonic plague is highly contagious, difficult to recognize and is frequently fatal. Therefore, the development of vaccines against this agent is crucial. Although tularemia is associated with low mortality, the highly infectious nature of aerosolized F. tularensis poses a substantive threat that is best met by vaccine development. Safer antitoxins and a vaccine are required to meet the threat of the use of botulinum toxin as a weapon of mass destruction. In this article, the current status of research in these areas is reviewed. PMID- 14554017 TI - Current trends in modern pharmaceutical analysis for drug discovery. AB - Traditionally, pharmaceutical analysis referred to the chemical analysis of drug molecules. However, over the years, modern pharmaceutical analysis has evolved beyond this to encompass combination techniques, high-throughput technologies, chemometrics, microdosing studies, miniaturization and nanotechnology. These analytical advances are now being employed in all stages of drug discovery and the focus of this review will be on how these technologies are being employed within this process. With new, improved and evolving technologies, as well as new applications for existing technology, the search for new drugs for the prevention and treatment of human diseases continues. PMID- 14554018 TI - Solid-state analysis of the active pharmaceutical ingredient in drug products. AB - The solid form of a drug substance is important when developing a new chemical entity. The crystalline form used in development is significant based on possible manufacturability, solubility, bioavailability and stability differences between the solid forms. Regulatory issues require that the form present in a solid dosage form or liquids containing undissolved drug substance be identified. Drug product samples can be analyzed by a variety of techniques to determine the crystal form present or changes that occur during the manufacture of a drug product. The form present will affect development, regulatory and intellectual property issues. PMID- 14554068 TI - Composite-composite repair bond strength: effect of different adhesion primers. AB - OBJECTIVES: Recently, new products have been introduced to repair composite restorations that may be used as 'one-step' primers or monomers and silane compounds which are used separately as 'multi-step' primers. The aim of this study was to compare the shear bond strength of the new composite resin to aged composite, by using different adhesion primers. METHODS: The substrates were particulate filler composite (Z250, 3M-ESPE), which was aged by boiling for 8 h and storing at 37 degrees C in water for 3 weeks. The aged substrate surfaces were wet-ground flat with 320-grit silicon carbide paper and subjected randomly (n=8) to either one-step adhesion primer: Compoconnect (CC) (Heraus Kulzer), or multi-step: Clearfil Repair (CF) (Kuraray) or an intermediate resin: Scothchbond Multi-purpose adhesive resin (3M-ESPE) according to the manufacturers' recommendations. Specimens with no surface treatment were used as control (C). New composite resin (Z250) was added to the substrate using 2 mm layer increments and light cured. The specimens were either water stored for 48 h or water stored for 24 h and then thermocycled for 6000 cycles. The shear bond strengths were measured with a crosshead speed of 1.0 mm/min using a universal testing machine. Data were analysed by two-way ANOVA and Tukey's post-hoc tests (p=0.05). RESULTS: All surface treatment methods showed significant difference compared to control (p<0.05). CF showed higher bond strength than CC and MP (p<0.05). Storage condition did not show a significant difference (p>0.05) in bond strength values. CONCLUSION: It was concluded that multi-step adhesion primer yielded higher bond strength compared to one-step primer or intermediate resin. PMID- 14554069 TI - Clinical evaluation of a self-etching and a one-bottle adhesive system at two years. AB - OBJECTIVES: The clinical performances of a self-etching adhesive system, Clearfil SE Bond, and a one-bottle adhesive system, Prime&Bond NT, were evaluated in non carious Class V restorations for a period of two years. METHODS: Ninety-eight restorations were made by one operator for 32 patients. The resin composite used to restore the teeth were Clearfil AP-X and Spectrum TPH for Clearfil SE Bond and Prime&Bond NT, respectively. Two clinicians at the baseline, 6th, 12th and 24th months evaluated the posterior composites according to the modified Ryge criteria's. For this, color match, marginal discoloration, marginal adaptation, recurrent caries, anatomic form, postoperative sensitivity and retention rates were considered. The changes across time and across groups were evaluated statistically. RESULTS: At two years, 88 restorations were reviewed in 28 patients. The retention rates for Clearfil SE Bond were 93 and 91% for Prime&Bond NT. The percentages of the retention rates of both adhesive systems were not found to be different when calculating the failure rates. Recurrent caries, anatomic form and postoperative sensitivity were scored as Alpha for all restorations. Two cases of both adhesive systems showed slight marginal discoloration problems. Three restorations of Prime&Bond NT and one of Clearfil SE Bond had marginal adaptation problems at two years. One case for each adhesive system had slight color change after the same period. CONCLUSION: We can conclude that both adhesive systems tested exhibited very good clinical performance at the end of two years. PMID- 14554070 TI - Association between local mechanical factors and orofacial pain: survey in the community. AB - OBJECTIVES: The aetiology of orofacial pain (OFP) is not well understood. We aimed to determine the relationship between OFP and local mechanical factors in an unselected general population sample. METHODS: A cross-sectional population based survey was conducted in the United Kingdom, involving 2504 participants (adjusted participation rate 74%). Postal questionnaire was used to collect information on OFP and local mechanical factors. RESULTS: A significant association was found between OFP and a history of tooth grinding, facial trauma, the jaw getting stuck or locked, a clicking or grating sound in the jaw joint when opening or closing the mouth, difficulty in opening the mouth wide, and chewing of pens or biting finger nails. The jaw getting stuck or locked had the highest relative risk of 2.7 (95% CI: 2.3-3.2). A history of orthodontic treatment, having any type of dentures and using chewing gum were not associated with OFP. There was some evidence of heterogeneity between types of OFP and local mechanical factors. CONCLUSIONS: Local factors play an important role in the aetiology of OFP. PMID- 14554071 TI - Changes in enamel surface roughness and adhesion of Streptococcus mutans to enamel after vital bleaching. AB - OBJECTIVES: The purpose of this in vitro study was to observe the influence of vital bleaching on changes to the enamel surface and adhesion of Streptococcus mutans to tooth enamel. METHODS: The coronal part of each of 70 extracted third molars was cut in half, with either the buccal or lingual half used for experiments or controls. Experimental halves were assigned to the following conditions: (A) enamel was bleached 1, 3 or 5 times using a bleaching material with or without etching; or (B) etched condition without bleaching. All control samples were kept intact in physiological saline solution. Surface roughness (Ra; roughness center-line average: microm) of enamel was measured for 35 pairs of specimens. TS broth culture medium containing 3% glucose was inoculated with S. mutans and cultured for 72 h before adding the other 35 pairs of specimens. Under scanning electron microscopy, the number of S. mutans colonies was counted and statistically analysed. RESULTS: Compared to controls, bleached enamel displayed increased colonies of S. mutans. Repeated bleaching further increased bacterial adhesion and maximal colonies counts were found under conditions of five bleaching treatments plus etching (p75% of the value-of-information attainable regarding the chicken-meat import decision. PMID- 14554142 TI - Assessment of veterinary needs of ruminant livestock owners in Ghana. AB - We assessed the needs of ruminant-livestock owners in three districts in Ghana for veterinary services, the acceptability of particular services or activities and the willingness of people to use services through public or private delivery systems. Purposive sampling was used in choosing 516, 100 and 100 ruminant livestock owners in East Mamprusi, Savelugu-Nanton and Tamale districts, respectively, for the administration of a questionnaire. The response proportions were 87-99%. Focus-group discussions (FGDs) were done in Savelugu-Nanton and Tamale districts. Animal diseases, housing, and lack of knowledge on management practices were identified as the three most important problems in the districts. Generally, accessibility to veterinary services and drugs was not easy. Large proportions of respondents identified advice on health, bathing or spraying against ectoparasites, castration, deworming, treatment involving injectables, sale of medicines or drugs, treatment for skin diseases, vaccinations, and treatment of wounds as activities that they needed. Many used veterinary staff to meet their needs and were willing to use the services of private veterinary providers if private clinics were established in their localities. PMID- 14554143 TI - The development and validation of an antibody-ELISA to detect Trypanosoma evansi infection in cattle in Australia and Papua New Guinea. AB - Trypanosoma evansi is exotic to Australia and Papua New Guinea (PNG). However, it might have been introduced to Papua (Indonesia); thus, there is a risk of it entering PNG and thence Australia. Because of logistical difficulties in PNG and northern Australia, surveillance for T. evansi must rely on serological tests. The accuracy of an Ab-ELISA using a detergent extract of T. evansi and three antigen fractions purified from the detergent extract using stepwise precipitation with saturated ammonium sulphate (AS) were compared. The ELISA using the AS 40-50% fraction had greater discriminatory power compared to the ELISA using the other antigen fractions. This ELISA then was compared with two commercial tests: the Card Agglutination Test for trypanosomiasis/T. evansi (CATT) and Suratex. CATT/T. evansi at 1/4 serum dilution has higher sensitivity and the ELISA has higher specificity. There is no likely benefit in combining antibody detection tests to improve the accuracy of diagnosis. Furthermore, the combination of Suratex (which was independent of the antibody tests) with the CATT or the ELISA did not improve the sensitivity. None of the tests was sufficiently sensitive to be used confidently to determine freedom from infection in animals imported into Australia from countries where T. evansi infection is endemic. PMID- 14554144 TI - Risk factors for porcine post-weaning multisystemic wasting syndrome (PMWS) in 149 French farrow-to-finish herds. AB - A cross-sectional study involving 149 farms was carried out in France in 2000 and 2001 to assess the risk factors for post-weaning multisystemic wasting syndrome (PMWS). The farms were divided into three groups according to their current or past PMWS status: CASES (current and typical PMWS), CONTROLS#1 (PMWS-free farms), and CONTROLS#2 (farms which have recovered from PMWS). Two different comparisons were tested: CASES versus CONTROLS#1 and CASES versus CONTROLS#2. In the first comparison, the odds of PMWS were increased when fattening pigs tested positive for parvovirus (PPv) and porcine reproductive and respiratory syndrome (PRRS) virus (OR=4.4 and 6.5, respectively), when separate vaccines for parvovirus and Erysipela for the gilts versus associated vaccines were used (OR=2.5), and when on-farm semen collection was used versus all the semen purchased from an insemination centre (OR=4.6). Large pens in weaning facilities increased the odds of PMWS (OR=4.1); whereas long empty periods in weaning and farrowing facilities versus shorter (OR=0.2), regular treatment against external parasites (OR=0.1), and housing the sows in collective pens during pregnancy versus individual pens (OR=0.3) all decreased the odds of PMWS. The same kinds of risk factors were found with the second comparison with, in addition, a common pit for several adjacent fattening rooms versus separate pits (OR=6.7) and a high level of cross fostering (OR=5.1). On the other hand, when farms had a self-replacement scheme for the gilts (OR=0.1), and when vaccination of the sows against E. coli was in place (OR=0.2), the odds of PMWS were decreased. PMID- 14554146 TI - Post-genomic technologies--thinking beyond the hype. PMID- 14554145 TI - Incidence of canine rabies in N'Djamena, Chad. AB - This work describes for the first time the incidence risk of passively reported canine rabies, and quantifies reported human exposure in N'Djamena (the capital of Chad). To diagnose rabies, we used a direct immunofluorescent-antibody test (IFAT). From January 2001 to March 2002, we were brought 34 rabies cases in dogs and three cases in cats. Canine cases were geographically clustered. The annual incidence risk of canine rabies was 1.4 (95% CI: 1.2, 1.7) per 1000 unvaccinated dogs. Most of the rabid dogs were owned-although free-roaming and not vaccinated against rabies. Most showed increased aggressiveness and attacked people without being provoked. Eighty-one persons were exposed to rabid dogs and four persons to rabid cats (mostly children<15 years old). Most of the exposed persons were neighbours or family members of the animal owner. Most exposures were transdermal bites, but nearly half of all exposed persons did not apply any first wound care or only applied a traditional treatment. In N'Djamena, humans are often exposed to canine rabies but do not use the full-course post-exposure treatment and wound care is insufficient. Most rabid dogs would be accessible to parenteral vaccination. Pilot vaccination campaigns are needed to determine the success of dog mass vaccination in N'Djamena as a way to prevent animal and human rabies. PMID- 14554147 TI - Cell cycle independent activity of cyclin D1. PMID- 14554148 TI - Minding the Ps and Qs of genomewide analysis. PMID- 14554149 TI - Infection stage clues to new antimalarial medicines. PMID- 14554152 TI - Science with a smile--cartoon capers. PMID- 14554153 TI - Quantitative image analysis: software systems in drug development trials. PMID- 14554154 TI - Ultra high quality uHTS. PMID- 14554155 TI - Structure-guided applications in drug discovery. PMID- 14554156 TI - In silico prediction of blood-brain barrier permeation. AB - This review examines the progress that is being made towards the in silico prediction of brain permeation. Following a brief introduction to the blood-brain barrier, the datasets currently available for in silico modeling are discussed. Recent developments in in silico models of brain permeation are summarized in the context of the current state of the art in prediction accuracy. An analysis of recent models is presented, focusing on what such models reveal about the molecular properties that determine brain permeation. The review concludes by presenting the current key issues in this area of research, noting in particular, the paucity of brain permeation data available for modeling. Finally, possible future directions are suggested. PMID- 14554157 TI - Vaccine adjuvants: role and mechanisms of action in vaccine immunogenicity. AB - Inactivated vaccines require adjuvants to stimulate an immune response. The choice of adjuvant or immune enhancer determines whether the immune response is effective, ineffective or damaging. Accordingly, there is a need for new adjuvants that stimulate the appropriate immunity, for example, T cell immunity for intracellular pathogens and cancer vaccines. In several adjuvants, the identification of chemical groups that interact with specific cell toll-like receptors (innate immunity) or receptors for co-stimulatory ligands (adaptive immunity), has enabled the establishment of structure-function relationships that are useful in the design of new adjuvants. Because of the crucial immunomodulating role of adjuvants, sub-unit vaccine development will remain dependent on new adjuvants. PMID- 14554158 TI - New approaches to in vitro models of blood-brain barrier drug transport. AB - The pharmaceutical industry has been searching for an in vitro blood-brain barrier (BBB) model that preserves in vivo transporter functions in CNS drug discovery and development. The application of conditionally immortalized cell lines derived from transgenic animals harboring temperature-sensitive SV40 large T-antigen gene, is a rational and promising approach to such a workable in vitro BBB model. The established brain capillary endothelial cell lines retain the in vivo transport rate of several compounds and various forms of gene expression. Furthermore, this new approach has enabled the development of stable and reproducible co-culture models with a pericyte cell line and/or an astrocyte cell line. PMID- 14554160 TI - Application of the SAXS method and viscometry for determination of the thickness of adsorbed polymer layers at the ZrO2-polymer solution interface. AB - The authors studied the influence of the molecular weight of polyacrylic acid (PAA) and polyacrylamide (PAM), solution pH and ionic strength, and the background electrolyte type on adsorption and the thickness of polyelectrolyte adsorption layers formed on ZrO(2) surface. Carboxyl groups distributed along PAA and PAM chains were shown to be responsible for their interface conformation, which directly influences the thickness of the adsorbed polyelectrolyte layers. Bonding of macromolecules with solid surface occurs through the hydrogen bridges of these groups. Two methods were applied to determine the PAA and PAM adsorption layer thickness on ZrO(2), i.e., SAXS (small angle X-ray scattering) and viscometry. Despite some limitations of the SAXS method resulting from the relationship between the size of solid pores, polymer molecular weight, and conformation of the adsorbed macromolecule, all obtained SAXS results were very close to those calculated from viscometry data. PMID- 14554161 TI - Synergism and foaming properties in mixed nonionic/fatty acid soap surfactant systems. AB - The synergism and foaming behavior of a mixed surfactant system consisting of a nonionic surfactant (polyethoxylated alkyl ether C(n)E(m)) and a fatty acid soap (sodium oleate) were studied. The micellar interaction parameter (the beta parameter) was determined from the cmc following the approach of Rubingh's regular solution theory. For both the C(12)E(6)/sodium oleate and the C(14)E(6)/sodium oleate mixtures, the results indicate a fairly strong attractive interaction (negative beta-values), which were in agreement with previous data reported for other nonionic/anionic surfactant systems. The characteristics of the foam produced from the surfactants were evaluated using a glass column equipped with a series of electrodes measuring the conductance of the foam, which enabled the water content of the foam to be determined. From these measurements, since the total foam volume was almost the same for all concentrations and surfactants, we compared the amount of liquid in the foam produced under dynamic foaming and the ability of the foam to entrain the liquid after the airflow was switched-off (static foam stability). The amount of liquid in the foam 100 s after the air was switched-off followed the order NaOl > C(12)E(6) > C(14)E(6). Also, the mixtures had the same foam volumes as the pure surfactants at the same concentration. However, both mixtures had higher concentrations of liquid in the foam when the mole fraction of the nonionic surfactant in the mixed surfactant system was greater than about >0.3 in the solution. PMID- 14554162 TI - Interfacial interactions between poly[L-lysine]-based branched polypeptides and phospholipid model membranes. AB - The interaction of five poly[L-lysine]-derived branched chain polypeptides of poly[Lys(X(i))] (X(i)K) or poly[Lys(X(i)-DL-Ala(m))] (XAK) with lipid bilayers (DPPC and DPPC/PG, 8:2) was studied by fluorescence polarization techniques. Two fluorescent probes, DPH and TMA-DPH, were utilized to monitor changes of motion in the internal and/or in the polar head regions, respectively. Results indicate that the interaction of polypeptides with neutral (DPPC) bilayers is mainly dependent on the polarity and electrical charge of side chains. The amphoteric E(i)K shows the highest level of interaction. Polycationic polypeptides (H(i)K, P(i)K, TAK) have a relatively small effect on the transition temperature of the lipids, while the polyanionic Succ-EAK has no effect at the alkyl chain region of the bilayer. Data with TMA-DPH indicate the lack of pronounced interaction between the polypeptides and the outer surface of the liposome. Similar tendency was documented for DPPC/PG vesicles. Polypeptides, H(i)K, and P(i)K induce significant changes in the transition temperature, thus indicating their insertion into the hydrophobic core of the bilayer without marked effect on the polar head region. Results suggest that these polypeptides (except E(i)K) have no destabilizing effect on liposomes studied. These properties are considered as beneficial for their use as safe carriers for bioactive molecules. PMID- 14554163 TI - Comparative study on Np(V) sorption on oxides of aluminum and silicon: effects of humic substance and carbonate in solution. AB - The sorption of Np(V) (total concentration 10(-5) mol/L) onto alumina and silica was studied by a batch technique under ambient aerobic conditions at 25 degrees C. The effects of pH, ionic strength, humic substance (HS), and added carbonate in aqueous solutions on the sorption of Np(V) onto alumina and silica were investigated. The sorption isotherms of Np(V) on alumina and the relationships between the equilibrium concentrations after sorption onto silica and the initial concentration before sorption in the absence and presence of HS and added carbonate in solutions were determined. It was found that as compared with the sorption of Np(V) onto alumina, the sorption by silica on the basis of mass is tremendously less, negative sorption of Np(V) onto silica occurs, the relative rate of sorption onto silica is quicker the sensitivity of sorption onto silica to ionic strength is higher, the pH dependence of sorption onto silica is less, and consequently the characteristics of Np(V) sorption onto alumina and silica are distinctly different. The effect of addition of HS or carbonate in solution was studied. Little effect of addition of HS (20 mg/L) on sorption onto alumina and silica were found. The addition of carbonate (0.001 mol/L) increased Np(V) sorption onto silica at pH values below 10 and decreased it at pH values above 10. PMID- 14554164 TI - Removal of methyl violet from aqueous solution by perlite. AB - The use of perlite for the removal of methyl violet from aqueous solutions at different concentration, pH, and temperature has been investigated. Adsorption equilibrium is reached within 1 h. The capacity of perlite samples for the adsorption of methyl violet was found to increase with increasing pH and temperature and decrease with expansion and increasing acid-activation. The adsorption isotherms are described by means of the Langmuir and Freundlich isotherms. The adsorption isotherm was measured experimentally at different conditions and the experimental data were correlated reasonably well by the adsorption isotherm of Langmuir. The order of heat of adsorption corresponds to a physical reaction. It is concluded that the methyl violet is physically adsorbed onto the perlite. The removal efficiency (P) and dimensionless separation factor (R) have shown that perlite can be used for removal of methyl violet from aqueous solutions, but unexpanded perlite is more effective. PMID- 14554165 TI - Adsorption of sodium dodecyl benzenesulfonate onto coal. AB - The adsorption behavior of sodium dodecyl benzenesulfonate (NaDDBS) on a raw (as received) coal sample and its demineralized variety with 11.3% and 1.2% ash contents respectively has been studied. The samples have been characterized by their proximate analysis, particle size distribution, surface area, porosity, density, points of zero charge, etc. Adsorption of NaDDBS on these two samples has been studied as a function of concentration of NaDDBS, temperature, pH, and presence of indifferent electrolyte in the medium. It has been observed that the isotherm exhibits two adsorption plateaus below and above the critical micelle concentration (CMC) of NaDDBS. Low heats of adsorption suggest weak hydrophobic bonding between adsorbent and adsorbate. The present work aims to correlate the adsorption of surfactant onto coal particles with the rheological behavior of coal-water slurry (CWS). The results reveal that addition of a very small amount of NaDDBS (0.3 wt% of coal) to 60% (w/w) CWS results in a marked reduction of the apparent viscosity of the CWS at a shear rate of 100 s(-1). The effect of pH on the apparent viscosity of CWS with and without the presence of the surfactant is also investigated. PMID- 14554166 TI - Modeling the adsorption of citric acid onto Muloorina illite and related clay minerals. AB - The adsorption of citric acid onto goethite, kaolinite, and illite was measured as a function of pH (adsorption edges) and concentration (adsorption isotherms) at 25 degrees C. The greatest adsorption was onto goethite and the least onto illite. Adsorption onto goethite was at a maximum below pH 5 and decreased as the pH was increased to pH 9. For kaolinite, maximum adsorption occurred between pH 4.5 and pH 7, decreasing below and above this pH region, while for illite maximum adsorption occurred between about pH 5 and pH 7, decreasing at both lower and higher pH. ATR-FTIR spectra of citrate adsorbed to goethite at pH 4.6, pH 7.0, and pH 8.8 were compared with those of citrate solutions between pH 3.5 and pH 9.1. While the spectra of adsorbed citrate resembled those of the fully deprotonated solution species, there were significant differences. In particular the C[bond]O symmetric stretching band of the adsorbed species at pH 4.6 and 7.0 changed shape and was shifted to higher wave number. Further spectral analysis suggested that citrate adsorbed as an inner-sphere complex at pH 4.6 and pH 7.0 with coordination to the surface most probably via one or more carboxyl groups. At pH 8.8 the intensity of the adsorbed bands was much smaller but their shape was similar to those from the deprotonated citrate solution species, suggesting outer-sphere adsorption. Insufficient citric acid adsorbed onto illite or kaolinite to provide spectroscopic information about the mode of adsorption onto these minerals. Data from adsorption experiments, and from potentiometric titrations of suspensions of the minerals in the presence of citric acid, were fitted by extended constant-capacitance surface complexation models. On the goethite surface a monodentate inner-sphere complex dominated adsorption below pH 7.9, with a bidentate outer-sphere complex required at higher pH values. On kaolinite, citric acid adsorption was modeled with a bidentate outer-sphere complex at low pH and a monodentate outer-sphere complex at higher pH. There is evidence of dissolution of kaolinite in the presence of citric acid. For illite two bidentate outer-sphere complexes provided a good fit to all data. PMID- 14554167 TI - Study of the factors affecting the forward and back extraction of yeast-lipase and its activity by reverse micelles. AB - The experimental results of yeast-lipase extracted by AOT-reverse-micelles in isooctane are reported. The effects of pH values and ionic strength in the aqueous phase, surfactant concentration, phase volume ratio, temperature, stirring time, and cosolvent concentration on the phase transfer of the lipase by the reverse micelles are studied. One hundred percent of forward extraction yield, 68% of back extraction yield, and 45% of activity recovery yield for the lipase are obtained. The phase transfer of the lipase is controlled by electrostatic and hydrophobic interactions. PMID- 14554168 TI - Determining the size distribution of core-shell spheres and other complex particles by laser diffraction. AB - The goal of this work is to determine the size distribution of hollow glass spheres by laser diffraction, an experiment which involves measuring angle dependent scattering of light from particles dispersed in a liquid. The proprietary software supplied with commercial instruments is not strictly applicable to our two-layer, glass-shell, hollow-core spheres because it requires that the particles have spatially homogeneous properties. We therefore developed Fortran code to compute the scattering from core-shell spherical particles. The results show that the scattering from representative hollow glass particles diverges from homogeneous sphere scattering when the radius decreases from 10 to 3 microm. Additionally, scattering measurements on two core-shell hollow glass powders were analyzed using the exact core-shell optical model and homogeneous sphere approximations. In both cases, the size distribution determined using the exact core-shell model differs from that determined using the homogeneous-sphere approximation when the distribution covers radii smaller than about 10 microm, as expected. The size distribution based on the exact core-shell optical model was determined using a new algorithm. Although the basic equations used in the algorithm have been published previously, they are developed here in a different form, which can be implemented using Fortran and MatLab routines available commercially and in the public domain. This algorithm could be used to determine the size distribution of other kinds of particles, such as cylindrical rods, as long as their angle-dependent scattering could be computed. PMID- 14554169 TI - Synthesis and stabilization of cadmium and thallium metal nanoparticles in a polymer matrix. AB - In situ synthesis of fine thallium and cadmium particles has been carried out by gamma-irradiation at room temperature in a preorganized polyacrylamide gel. It appears that Cd and Tl ions are chelated by the polymer matrix through physical entrapment as well as by weak chemical complexation. The metallic nanoparticles of Cd or Tl are well dispersed in the polymer matrix. The nanoparticles are stable in air and transfer electrons to methyl viologen (paraquat; 1,1'-dimethyl 4,4'-dipyridinium dichloride; MV(2+)(Cl(-))(2)). The radical, MV(+.), formed upon electron transfer, also gets trapped in the polymer matrix and shows high stability even in the presence of air. PMID- 14554170 TI - Liquid-liquid phase-transfer of magnetic nanoparticles in organic solvents. AB - We report a novel route for the preparation of well-defined colloidal dispersions of magnetic nanoparticles stabilized by steric repulsion in organic solvents. The usual methods standardly lead to the surfaction of multiparticle aggregates, incompatible with our long-term aim of studying and modeling the influence of magnetic dipolar interactions in colloidal dispersions which are free of aggregates, all other interactions being perfectly defined. A new and reproducible method based on a surfactant-mediated liquid-liquid phase transfer of individually dispersed gamma-Fe(2)O(3) nanoparticles from an aqueous colloidal dispersion to an organic phase is developed. The choice of the reagent and the preparation techniques is discussed. Among several solvent/surfactant pairs, the cyclohexane/dimethyldidodecylammonium bromide (DDAB) system is found to fulfill the colloidal stability criterion: aggregation does not appear, even upon aging. A complete transfer of isolated particles is observed above a threshold in DDAB concentration. The nanoparticle surface is then fully covered with adsorbed DDAB molecules, each surfactant head occupying a surface of 0.57+/-0.05 nm(2). The volume fraction of the cyclohexane-based organosols is easily tunable up to a volume fraction of 12% by modifying the volume ratio of the organic and of the aqueous phases during the liquid-liquid phase transfer. PMID- 14554171 TI - Filler-elastomer interactions: influence of silane coupling agent on crosslink density and thermal stability of silica/rubber composites. AB - In this work, the crosslink density and thermal stability of the silica/rubber composites treated by silane coupling agents, i.e., gamma-aminopropyl triethoxysilane (APS), gamma-chloropropyl trimethoxysilane (CPS), and gamma methacryloxypropyl trimethoxysilane (MPS), were investigated. The chemical structures of modified silicas were studied in term of solid-state 29Si NMR spectroscopy. The crosslink density of the composites was determined by swelling measurement. The development of organic functional groups on silica surfaces treated by coupling agents led to an increase in the crosslink density of the composites, resulting in increasing final thermal stability of the composites. The composites treated by MPS showed the superior crosslink density and thermal stability in these systems. The results could be explained by the fact that the organic functional groups of silica surfaces by silane surface treatments led to an increase of the adhesion at interfaces between silicas and the rubber matrix. PMID- 14554172 TI - Surface patterning via evaporation of ultrathin films containing nanoparticles. AB - The dewetting dynamics of ultrathin films containing potentially surface-active nanoparticles is considered in the presence of evaporation. Evolution equations for the film height and particle surface and bulk concentration are derived using a lubrication model coupled by a constitutive relation for the dependence of the viscosity on local particle concentration. A linear stability analysis and numerical simulations are used to determine how particle mass distribution depends on the various physical parameters such as equilibrium film separation distance, initial packing concentration, rate of evaporation, and particle surface activity. Our results show that when starting from an initially uniform distribution the particles become aligned into distinct "bands" in rectilinear geometry, or "rings" in cylindrical geometry. The functional dependence of the pattern spacing on relevant system parameters is studied and detailed herein. PMID- 14554173 TI - Influence of solution composition and ultrasonic treatment on optical spectra of TiO2 aqueous suspensions. AB - Investigation of TiO(2) aqueous suspensions has shown that their optical spectra can be unstable, with instability not related to precipitation or adherence of TiO(2) particles to the vessel walls. Increase of ionic strength of the suspension as well as neutralization of charged TiO(2) particles via pH adjustment accelerates the optical density drop. Vice versa, increasing the charge of TiO(2) particles via shifting pH in acidic or basic directions stabilizes the suspension's optical spectra, and ultrasonic treatment promotes optical density recovery. The observed behavior is attributed to alteration in the size of the suspension aggregates. PMID- 14554174 TI - Kinetic study of epoxy curing in the glass fiber/epoxy interface using dansyl fluorescence. AB - The fluorescence response of the dansyl chromophore has been used to study the kinetic of epoxy curing processes. With this new method, comparison between the curing at the interface of a glass fiber/epoxy and in the epoxy bulk of a composite material was studied. The effect of two glass fiber surface treatments was investigated. Commercial E-glass fibers were surface coated with 3 aminopropyltriethoxysilane (APTES) and 3-aminopropylmethyldiethoxysilane (APDES). Fluorimetry (using fluorescent labels) and FT-NIR (Fourier transformed infrared spectroscopy in the near range) techniques were used to monitor the curing process in these composite materials. From the analysis of the data obtained, different simple kinetic models were discussed and apparent activation energies were obtained. Furthermore, from those techniques the respective results were compared to obtain complementary information. Independently of the sample and the technique used for the kinetic analysis, no variation of the activation energy of the epoxy curing reaction was found, which suggests that there are no changes in the mechanism of the reaction along the process. Fluorescence from dansyl located at the glass fiber/epoxy interface reflected that the kind of reinforcement treatment clearly affects the epoxy curing process exactly in that region. However, when analytical response comes from the whole system the mechanism of the reaction does not seem to change with the silane coating used although is quite different in comparison with the process at the interface. PMID- 14554175 TI - Photocatalysis and hydrophilicity of doped TiO2 thin films. AB - TiO(2) thin films were prepared using the dip-coating method with a polymeric sol including additives such as Al, W, and Al+W to examine two major properties: photocatalysis and hydrophilicity. W-doped films showed the best photocatalytic efficiency, while Al-doped film was poorer than undoped samples. However, good hydrophilicity in terms of saturation contact angle and surface conversion rate was found in Al- and (Al+W)-mixed-doped films. It was found that deep electron hole traps and high surface acidity of W-doped TiO(2) thin film were the major factors in high photocatalytic efficiency. In addition, low surface acidities of Al- and (Al+W)-doped films provided better hydrophilicity than W-doped ones. However, the amount of [Ti(3+)] point defects on the surface was another major factor, probably the most important, in getting the best hydrophilicity. Conclusively, it seemed that many parts of the photocatalysis mechanism depend more on bulk-related properties than do those of hydrophilicity, which can be defined as an interfacial (surface) or near-surface-restricted process. PMID- 14554176 TI - On the electrochemical characterization of ion-selective cellulose acetate membranes with and without stigmasterol liquid membranes. AB - Stigmasterol, a plant product, has been used as a surfactant to generate liquid membranes supported on a cellulose acetate matrix. Electrochemical characterization of the membrane has been attempted with a view to simulating its behavior with natural membranes by measuring membrane potentials and membrane conductance. The selectivity of cellulose acetate membrane kept in contact with magnesium chloride solutions of different mean concentrations has been found to change from anion to cation. Transport numbers have been estimated from membrane potential data. Permselectivity and fixed charge density values for the cellulose acetate membrane with and without stigmasterol have been determined from transport numbers. The variation of these parameters with concentration and pH has also been examined. PMID- 14554177 TI - Porosity and interior structure of flocculated activated sludge floc. AB - This work estimated the porosities of activated sludge flocs, cationic polyelectrolyte flocculated, based on free-settling tests, buoyant weight measurements, and confocal laser scanning microscope (CLSM) tests. The extent of advective flow was estimated based on bubble-tracking test. The former two measurements suggested a close-to-unity porosity, that is, an extremely void floc interior. Meanwhile, the latter two tests recommended a dense floc interior with a porosity less than 64%. A discrepancy exists between the porosities estimated by various tests. A floc model was proposed based on the understanding that a vast amount of bound water in the floc was regarded as void in buoyant weight measurement, but was impermeable for advective flow. The distribution rather than the mean value of the porosity controls the advective flow. There existed no simple correlation between the porosities measured by different tests. PMID- 14554178 TI - Surface properties of aqueous amino acid solutions. I. Surface tension of hydrochloric acid-glycine and glycine-sodium hydroxide systems. AB - The surface tensions of aqueous solutions of four mixtures (hydrochloric acid glycine hydrochloride, glycine hydrochloride-glycine, glycine-sodium glycinate, and sodium glycinate-sodium hydroxide) were measured as a function of total molality and mole fraction. The measurements correspond to the change in surface tension with variation of pH. The contribution of glycine hydrochloride to the increments in surface tension is equivalent to that observed for the aqueous solution of glycine, while the contribution of sodium glycinate is much larger than that of glycine. The variations in surface tension on mixing in the surface region are discussed using comparisons with mixtures of simple salts. PMID- 14554179 TI - Testing the Gouy-Chapman theory by means of surface tension measurements for SDS NaCl-H2O mixtures. AB - Surface tension isotherms were measured for sodium dodecyl sulfate (SDS) at different concentrations of added salt (NaCl). The free energy of the surfactant monolayer was assessed by invoking the Gouy-Chapman theory for the charged head groups, the hydrophobic (Tanford) free energy of transfer of the hydrocarbon chain, and the hydrocarbon chain configurational free energy according to Gruen's calculations and finally macroscopic contact terms. In particular, the effect of an increased salt concentration in bulk was examined. Theoretical predictions compare well with the experimental findings, and good agreement was found with respect to both the variation of free energy of the monolayer and the surface pressure behavior. Thus, at least for a liquid-expanded monolayer of SDS, the Gouy-Chapman model yields a satisfactory account of the electrostatic contribution to the thermodynamic properties at different salt concentrations of NaCl. PMID- 14554180 TI - Shape change of prism solids in different liquid systems during local corrosion. AB - As reported in many previous publications, the cross section shape of prism solids will maintain the original square or become round after a certain duration of dipping and corrosion in certain melts. This paper gives theoretical explanations for these phenomena. Based on the deduction of the Laplace equation, it is found that for the solid-liquid system with contact angle (theta) smaller than 90 degrees, the maximum height of liquid surface on the lateral faces (H(f)) is greater than that at the prism edges (H(e)) before corrosion. If the corrosion and dissolution of the solid sample reduce the liquid surface tension, the case of H(f)>H(e) will be maintained or intensified due to the larger extent of dissolution at the edges, and the down-and-up motion of the liquid surface on the prism will mainly occur on the lateral faces, resulting in the maintenance of the original square shape of the sample's cross section. However, if the dissolution of the solid sample increases the liquid surface tension, the case H(f)90 degrees, the liquid surface height exhibits the opposite direction, and the evolution trends of the cross section shape are similar. PMID- 14554181 TI - Effect of counterions on surface and foaming properties of dodecyl sulfate. AB - The influence of counterions of surfactant on interfacial properties is studied by measuring foamability, foam stability, equilibrium and dynamic surface tension, and surface viscosity. The surfactant chosen is anionic dodecyl sulfate with various counterions, Li(+), Na(+), Cs(+), and Mg(++). Surface tension measurements show a decrease in the following order: LiDS > NaDS > CsDS > Mg(DS)(2). Foamability done using shaking method shows similar order as surface tension, i.e., LiDS > NaDS > CsDS > Mg(DS)(2). This has been explained in terms of the differences in micellar stability and diffusion of monomers. This is further confirmed by our dynamic surface tension results, which show the same order as equilibrium surface tension (i.e., LiDS > NaDS > CsDS > Mg(DS)(2)) at low bubble frequencies but the order is LiDS > NaDS = Mg(DS)(2) > CsDS at high bubble frequencies. Foam stability measurements were done at concentrations below and above cmc to elucidate the role of micelles. It was found that there is no significant change in foam stability when counterions are changed for surfactant concentration values below the cmc, but at concentration above cmc the foam stability of CsDS and Mg(DS)(2) are much greater than LiDS and NaDS indicating presence of stable micelles are essential to high foam stabilities. Surface viscosity measurements correlated well with the foam stability trends and gave the following order LiDS < NaDS < CsDS << Mg(DS)(2), indicating that the molecules of CsDS and Mg(DS)(2) are tightly packed at the air/water interface. PMID- 14554182 TI - Physicochemical properties of quaternary ammonium bromide-type trimeric surfactants. AB - Trimeric surfactants of quaternary ammonium bromide (m-2-m-2-m, where m is the hydrocarbon chain length of 8, 10, or 12) with three hydrocarbon chains and three hydrophilic groups connected by two ethylene spacer chains were synthesized by the reaction of N,N,N',N",N"-pentamethyldiethylenetriamine and the corresponding alkyl bromide. Their physicochemical properties were characterized by surface tension, static and dynamic light-scattering, and fluorescence spectrum of pyrene techniques. The critical micelle concentrations (cmc's) of m-2-m-2-m shifted to lower concentrations with increasing hydrocarbon chain length, and their values were smaller by about one to three orders of magnitude than those of the corresponding dimeric (m-2-m) and monomeric surfactants (C(m)TAB) with the same hydrocarbon chain length. Of these surfactants, 10-2-10-2-10 showed the greatest efficiency in lowering the surface tension and provided the smallest occupied area per molecule, indicating that it adsorbs more compactly at the air/water interface. In addition, from the static and dynamic light-scattering measurements, the aggregation numbers of the trimeric surfactants at the cmc were very small, and two hydrodynamic diameters above the cmc were observed. PMID- 14554183 TI - The solubilization of n-pentane gas in sodium dodecyl sulfate-polyethylene glycol solutions with and without electrolyte. AB - The solubility of n-pentane gas in aqueous solution of sodium dodecyl sulfate (SDS), SDS-0.1 wt% polyethylene oxide (PEG), SDS-0.1 wt% PEG+NaCl (0.1 mol/l), and SDS-0.1 wt% PEG+NaOH (0.1 mol/l) has been determined at 318.15 K. The concentration of SDS (m(SDS)) is up to 50 mmol/kg. The solubility increases linearly with the concentration of SDS above its critical micelle concentration (CMC) or critical aggregation concentration (CAC), indicating that micelles in the solutions solubilize the gas molecules and the solubility of n-pentane gas in the micelles is independent of the SDS concentration. It was found that the solubilization ability of micelles bound to PEG and free micelles to n-pentane gas is almost the same. The solubility of n-pentane gas in micelle phase is three magnitudes higher than that in the bulk solution. The solubilization property of SDS is changed by the addition of PEG, although the solubilizing effect of the polymer alone is not considerable. NaCl and NaOH affect the solubilization noticeably and increase the interaction strength between SDS and PEG. The standard Gibbs energies for the transfer of n-pentane gas from bulk phase to micelle phase are large negative values, indicating that the hydrocarbon gas prefers to exist in the hydrophobic interior of the micelles. PMID- 14554185 TI - Thermodiffusion in compact clays. AB - An experimental cell has been devised in order to measure the Soret coefficients in a compact clay, namely argilite, when a concentration difference of a binary mixture is applied simultaneously with a temperature difference. Temperature gradients have been imposed in the same direction as concentration gradients or in the opposite one. The sign of the Soret coefficients is related to the respective direction of these gradients. Generally, mass transfer was found to be enhanced by the Soret effect. Experimental values of the Soret coefficient are given. PMID- 14554184 TI - Aggregation and solubility behavior of asphaltenes and their subfractions. AB - Asphaltenes from four different crude oils (Arab Heavy, B6, Canadon Seco, and Hondo) were fractionated in mixtures of heptane and toluene and analyzed chemically, by vapor pressure osmometry (VPO), and by small angle neutron scattering (SANS). Solubility profiles of the asphaltenes and their subfractions indicated strong cooperative asphaltene interactions of a particular subfraction that is polar and hydrogen bonding. This subfraction had lower H/C ratios and modestly higher N, V, Ni, and Fe contents than the less polar and more soluble subfraction of asphaltenes. VPO and SANS studies indicated that the less soluble subfractions formed aggregates that were considerably larger than the more soluble subfractions. In general, asphaltene aggregate size increased with decreasing solvent aromaticity up to the solubility limit, beyond which the aggregate size decreased with heptane addition. The presence of a low wavevector Q feature in the scattering curves at 25 degrees C indicated that the individual aggregates were flocculating; however, the intensity of the feature was diminished upon heating of the samples to 80 degrees C. The solubility mechanism for Canadon Seco asphaltenes, the largest aggregate formers, appears to be dominated by aromatic pi-bonding interactions due to their low H/C ratio and low nitrogen content. B6 and Hondo asphaltenes formed similar-sized aggregates in heptol and the solubility mechanism is most likely driven by polar interactions due to their relatively high H/C ratios and high nitrogen contents. Arab Heavy, the least polar asphaltene, had a H/C ratio similar to Canadon Seco but formed the smallest aggregates in heptol. The enhancement in polar and pi-bonding interactions for the less soluble subfraction indicated by elemental analysis is reflected by the aggregate size from SANS. The less soluble asphaltenes contribute the majority of species responsible for aggregation and likely cause many petroleum production problems such as pipeline deposition and water-in-oil emulsion stabilization. PMID- 14554186 TI - Shear rate and viscosity corrections for a Casson fluid in cylindrical (Couette) geometries. AB - Shear rate and viscosity correction factors for a Casson fluid in cylindrical (Couette) geometries--type I, concentric cylinder with a narrow gap, and type II, cylinder in a semi-infinite medium--are developed in this work. The analytical correction factors compare well with published data corrected by general methods. Corrected viscosities were also compared to viscosity-shear rate data on chocolate in an absolute viscometer (cone-and-plate), which requires no corrections. This comparison showed excellent agreement of the corrected data in the higher shear rate range, but not very good agreement in the lower shear rate range. Reasons for this are discussed. In addition, a Reiner-Riwlin type formulation was developed for determination of model constants directly from rotational viscometry data without the need to correct for non-Newtonian shear rate, but this method is less accurate than existing slope-intercept methods for determining model constants. The analytical correction procedure developed here is expected to be useful wherever the Casson model is used to describe the rheology of a fluid, particularly one exhibiting yield-stress pseudoplastic behavior. PMID- 14554187 TI - Effect of chemical composition on electrokinetics of diaspore. AB - The effect of chemical composition of diaspores on their electrokinetics was studied. Increasing SiO(2) content in a diaspore sample was found to decrease its isoelectric point. The X-ray diffraction and SEM microanalysis showed the absence of distinct SiO(2) phases. A linear correlation was found to exist between the measured isoelectric point and alumina to silica mass ratio in diaspore samples. The linear regression analysis of the experimental data suggests a more significant impact of silicon content than aluminum content, indicating a preferential adsorption of dissolved silicon on diaspore samples and/or preferential dissolution of aluminum from diaspore samples. PMID- 14554188 TI - Experimental study of the immiscible displacement of shear-thinning fluids in pore networks. AB - The pore scale mechanisms and network scale transient pattern of the immiscible displacement of a shear-thinning nonwetting oil phase (NWP) by a Newtonian wetting aqueous phase (WP) are investigated. Visualization imbibition experiments are performed on transparent glass-etched pore networks at a constant unfavorable viscosity ratio and varying values of the capillary number (Ca), and equilibrium contact angle (theta(e)). Dispersions of ozokerite in paraffin oil are used as the shear-thinning NWP, and aqueous solutions of PEG colored with methylene blue are used as the Newtonian WP. At high Ca values, the tip splitting and lateral spreading of WP viscous fingers are suppressed; at intermediate Ca values, the primary viscous fingers expand laterally with the growth of smaller capillary fingers; at low Ca values, network spanning clusters of capillary fingers separated by hydraulically conductive noninvaded zones of NWP arise. The spatial distribution of the mobility of shear-thinning NWP over the pore network is very broad. Pore network regions of low NWP mobility are invaded through a precursor advancement/swelling mechanism even at relatively high Ca and theta(e) values; this mechanism leads to irregular interfacial configurations and retention of a substantial amount of NWP along pore walls; it becomes the dominant mechanism in displacements performed at low Ca and theta(e) values. The residual NWP saturation increases and the end WP relative permeability decreases as Ca increases and both become more sensitive to this parameter as the shear-thinning behavior strengthens. The shear-thinning NWP is primarily entrapped in individual pores of the network rather than in clusters of pores bypassed by the WP. At relatively high flow rates, the amplitude of the variations of pressure drop, caused by fluid redistribution in the pore network, increase with shear-thinning strengthening, whereas at low flow rates, the motion of stable and unstable menisci in pores is reflected in strong pressure drop fluctuations. PMID- 14554189 TI - Structure and rheological behavior of highly charged colloidal particles in a cylindrical pore I. Effect of pore size. AB - In this work we performed nonequilibrium Brownian dynamics (NEBD) computer simulations of highly charged colloidal particles in diluted suspension under a parabolic flow in cylindrical pores. The influence of charged and neutral cylindrical pores on the structure and rheology of suspensions is analyzed. A shear-induced disorder-order-disorder-like transition was monitored for low shear rates and small pore diameters. We calculate the concentration profiles, axial distribution functions, and axial-angular pair correlation functions to determine the structural properties at steady state for a constant shear flow for different pore sizes and flow strengths. Similar behavior has been observed in a planar narrow channel in the case of charged interacting colloidal particles (M.A. Valdez, O. Manero, J. Colloid Interface Sci. 190 (1997) 81). The mobility of the particles in the radial direction decreases rapidly with the flow and becomes practically frozen. The flow exhibits non-Newtonian shear thinning behavior due to interparticle interactions and particle-wall interaction; the apparent viscosity is lower as the pore diameter decreases, giving rise to an apparent slip in the colloidal suspension. The calculated slip velocity was higher than that obtained in a rectangular slit under shear flow. PMID- 14554190 TI - Hydrodynamic instability and coalescence in trains of emulsion drops or gas bubbles moving through a narrow capillary. AB - We investigate the effect of surfactant on the hydrodynamic stability of a thin liquid film formed between two emulsion drops or gas bubbles, which are moving along a narrow capillary. A ganglion (deformed drop or bubble in a pore) is covered by an adsorption monolayer of surfactant. Due to the hydrodynamic viscous friction, the surfactant is dragged from the front part of a moving ganglion toward its rear part. Consequently, the front and rear parts are, respectively, depleted and enriched in adsorbed surfactant. When such two ganglia move one after another, surfactant molecules desorb from the rear part of the first ganglion and are transferred by diffusion, across the intermediate liquid film, to the front part of the second ganglion. This leads to the appearance of a diffusion-driven hydrodynamic instability, which may cause coalescence of the two neighboring drops or bubbles. The coalescence occurs through a dimple-like perturbation in the film thickness, which is due to a local lowering in the pressure caused by a faster circulation of the liquid inside the film, which in turn is engendered by the accelerated surfactant diffusion across the thinner parts of the film. The developed theory predicts the critical distance between the two ganglia, which corresponds to the onset of coalescence, and its dependence on the radius of the capillary channel, velocity of motion, surfactant concentration and type of the operative surface forces. The results can be useful for a better understanding and quantitative description of the processes accompanying the flow of emulsions and foams though porous media. PMID- 14554191 TI - K+ fluxes in Schizosaccharomyces pombe. AB - All living cells accumulate high concentrations of K+ in order to keep themselves alive. To this end they have developed a great diversity of transporters. The internal level of K+ is the result of the net balance between the activities of the K+ influx and the K+ efflux transporters. Potassium fluxes have been extensively studied and characterized in Saccharomyces cerevisiae. However, this is not the case in the fission yeast and, in addition, the information available indicates that both yeasts present substantial and interesting differences. In this paper we have reviewed and summarized the information on K+ fluxes in Schizosaccharomyces pombe. We have included some unpublished results recently obtained in our laboratory and, in particular, we have highlighted the significant differences found between the well-known yeast S. cerevisiae and the fission yeast Sch. pombe. PMID- 14554192 TI - Sugar-induced apoptosis in yeast cells. AB - Sugars induce death of Saccharomyces cerevisiae within a few hours in the absence of additional nutrients to support growth; by contrast, cells incubated in water or in the presence of other nutrients without sugar remain viable for weeks. Here we show that this sugar-induced cell death (SICD) is characterized by rapid production of reactive oxygen species (ROS), RNA and DNA degradation, membrane damage, nucleus fragmentation and cell shrinkage. Addition of ascorbic acid to sugar-incubated cells prevents SICD, indicating that SICD is initiated by ROS. The lack of a protection mechanism against SICD suggests that sugars use to be the limiting nutrients for yeast and are probably depleted before all other nutrients. Being the limiting nutrient, sugars became the growth-stimulating agent, signaling the presence of sufficient nutrients for growth, but in the absence of the complementing nutrients they induce apoptotic death. PMID- 14554193 TI - Yeast as a tool to study Bax/mitochondrial interactions in cell death. AB - The budding yeast Saccharomyces cerevisiae has proven to be a powerful tool in investigations of the molecular aspects of the events involved in apoptosis, particularly the steps implicating mitochondria. Yeast does not have obvious homologs of the proteins involved in the regulation of apoptosis, and provides a simplified model system in which the function of these proteins can be unraveled. This review focuses on the interactions of two of the major pro-apoptotic Bcl-2 family members, Bax and Bid, with mitochondria. It is shown that yeast has allowed questioning of several crucial aspects of the function of these two proteins, namely the molecular mechanisms driving their insertion into the mitochondrial outer membrane and those leading to the permeabilization to cytochrome c. More recently, signaling pathways leading to Bax-induced cell death, as well as other forms of cell death, have been identified in yeast. Both 'apoptosis-like' and autophagy-related forms of cell degradation are involved, and mitochondria play a central role in these two signaling pathways. PMID- 14554194 TI - A Kluyveromyces lactis mutant in the essential gene KlLSM4 shows phenotypic markers of apoptosis. AB - We report the study of Kluyveromyces lactis cells expressing a truncated form of KlLSM4, a gene ortholog to LSM4 of Saccharomyces cerevisiae which encodes an essential protein involved in both pre-mRNA splicing and mRNA decapping. We had previously demonstrated that the first 72 amino acids of the K. lactis Lsm4p (KlLsm4Deltap) can restore cell growth in both K. lactis and S. cerevisiae cells not expressing the endogenous protein. However, cells showed a remarkable loss of viability in stationary phase. Here we report that cells expressing KlLsm4Deltap presented clear apoptotic markers such as chromatin condensation, DNA fragmentation, accumulation of reactive oxygen species, and showed increased sensitivity to different drugs. RNA analysis revealed that pre-mRNA splicing was almost normal while mRNA degradation was significantly delayed, pointing to this as the possible step responsible for the observed phenotypes. PMID- 14554195 TI - Genetic and biochemical interactions between the Arp2/3 complex, Cmd1p, casein kinase II, and Tub4p in yeast. AB - Arc35p, a component of the Arp2/3 complex, plays at least two distinct roles, regulating the actin cytoskeleton, but also microtubule function during cell division. Both functions involve calmodulin (CMD1). To investigate the pathway affecting microtubule function, we identified genes that are able to suppress the temperature-sensitive growth defect of the arc35-1 strain. Genes encoding gamma tubulin (TUB4) or any subunit of casein kinase II (CKII) suppressed this growth defect, but did not suppress the growth defect of a mutant in another subunit of the Arp2/3 complex, arp2-1. We could also show a physical association of Arc35p with subunits of CKII, Cmd1p, and Tub4p. Based on the exclusive localization of Arc35p to the cytosolic Arp2/3 complex and on mutant phenotypes, we propose that the role of the Arc35p/CKII interaction might be to activate a cytosolic pool of gamma-tubulin, likely via calmodulin, for its nuclear and/or cytoplasmic functions. PMID- 14554196 TI - Isolation and characterization of the HO gene from the yeast Saccharomyces paradoxus. AB - A DNA fragment homologous to the homothallism (HO) gene of Saccharomyces cerevisiae was isolated from Saccharomyces paradoxus and was found to contain an open reading frame that was 90.9% identical to the coding sequence of the S. cerevisiae HO gene. The putative HO gene was shown to induce diploidization in a heterothallic haploid strain from S. cerevisiae. Phylogenetic analysis revealed that the coding and 5'-upstream regulatory regions from five Saccharomyces sensu stricto HO genes have coevolved, and that S. paradoxus is phylogenetically closer to S. cerevisiae than to S. bayanus. Finally, heterothallic haploid strains were isolated from the original homothallic type strain of S. paradoxus by disrupting the S. paradoxus HO gene with the S. cerevisiae URA3 gene. PMID- 14554197 TI - Aerobic physiology of redox-engineered Saccharomyces cerevisiae strains modified in the ammonium assimilation for increased NADPH availability. AB - Recombinant strains altered in the ammonium assimilation pathways were constructed with the purpose of increasing NADPH availability. The NADPH dependent glutamate dehydrogenase encoded by GDH1, which accounts for a major fraction of the NADPH consumption during growth on ammonium, was deleted, and alternative pathways for ammonium assimilation were overexpressed: GDH2 (NADH consuming) or GLN1 and GLT1 (the GS-GOGAT system). The flux through the pentose phosphate pathway during aerobic growth on glucose decreased to about half that of the reference strain Saccharomyces cerevisiae CEN.PK113-7D, indicating a major redox alteration in the strains. The basic growth characteristics of the recombinant strains were not affected to a great extent, but the dilution rate at which the onset of aerobic fermentation occurred decreased, suggesting a relation between the onset of the Crabtree effect and the flux through the Embden-Meyerhof Parnas pathway downstream of glucose 6-phosphate. No redox effect was observed in a strain containing a deletion of GLR1, encoding glutathione reductase, an enzyme that is NADPH-consuming. PMID- 14554198 TI - High-level functional expression of a fungal xylose isomerase: the key to efficient ethanolic fermentation of xylose by Saccharomyces cerevisiae? AB - Evidence is presented that xylose metabolism in the anaerobic cellulolytic fungus Piromyces sp. E2 proceeds via a xylose isomerase rather than via the xylose reductase/xylitol-dehydrogenase pathway found in xylose-metabolising yeasts. The XylA gene encoding the Piromyces xylose isomerase was functionally expressed in Saccharomyces cerevisiae. Heterologous isomerase activities in cell extracts, assayed at 30 degrees C, were 0.3-1.1 micromol min(-1) (mg protein)(-1), with a Km for xylose of 20 mM. The engineered S. cerevisiae strain grew very slowly on xylose. It co-consumed xylose in aerobic and anaerobic glucose-limited chemostat cultures at rates of 0.33 and 0.73 mmol (g biomass)(-1) h(-1), respectively. PMID- 14554199 TI - Metabolic engineering of Saccharomyces cerevisiae for the synthesis of the wine related antioxidant resveratrol. AB - The stilbene resveratrol is a stress metabolite produced by Vitis vinifera grapevines during fungal infection, wounding or UV radiation. Resveratrol is synthesised particularly in the skins of grape berries and only trace amounts are present in the fruit flesh. Red wine contains a much higher resveratrol concentration than white wine, due to skin contact during fermentation. Apart from its antifungal characteristics, resveratrol has also been shown to have cancer chemopreventive activity and to reduce the risk of coronary heart disease. It acts as an antioxidant and anti-mutagen and has the ability to induce specific enzymes that metabolise carcinogenic substances. The objective of this pilot study was to investigate the feasibility of developing wine yeasts with the ability to produce resveratrol during fermentation in both red and white wines, thereby increasing the wholesomeness of the product. To achieve this goal, the phenylpropanoid pathway in Saccharomyces cerevisiae would have to be introduced to produce p-coumaroyl-CoA, one of the substrates required for resveratrol synthesis. The other substrate for resveratrol synthase, malonyl-CoA, is already found in yeast and is involved in de novo fatty-acid biosynthesis. We hypothesised that production of p-coumaroyl-CoA and resveratrol can be achieved by co-expressing the coenzyme-A ligase-encoding gene (4CL216) from a hybrid poplar and the grapevine resveratrol synthase gene (vst1) in laboratory strains of S. cerevisiae. This yeast has the ability to metabolise p-coumaric acid, a substance already present in grape must. This compound was therefore added to the synthetic media used for the growth of laboratory cultures. Transformants expressing both the 4CL216 and vst1 genes were obtained and tested for production of resveratrol. Following beta-glucosidase treatment of organic extracts for removal of glucose moieties that are typically bound to resveratrol, the results showed that the yeast transformants had produced the resveratrol beta-glucoside, piceid. This is the first report of the reconstruction of a biochemical pathway in a heterologous host to produce resveratrol. PMID- 14554200 TI - Combined overexpression of genes of the ergosterol biosynthetic pathway leads to accumulation of sterols in Saccharomyces cerevisiae. AB - Genes of the post-squalene ergosterol biosynthetic pathway in Saccharomyces cerevisiae have been overexpressed in a systematic approach with the aim to construct yeast strains that produce high amounts of sterols from a squalene accumulating strain. This strain had previously been deregulated by overexpressing a truncated HMG-CoA reductase (tHMG1) in the main bottleneck of the early ergosterol pathway. The overexpression of the gene ERG1 (squalene epoxidase) induced a significant decrease of the direct substrate squalene, a high increase of lanosterol, and a small increase of later sterols. The overexpression of the ERG11 gene encoding the sterol-14alpha-demethylase resulted in a decrease of lanosterol and an increase of downstream sterols. When these two genes were simultaneously overexpressed, later sterols from zymosterol to ergosterol accumulated and the content of squalene was decreased about three fold, indicating that these steps had limited the transformation of squalene into sterols. The total sterol content in this strain was three-fold higher than in a wild-type strain. PMID- 14554201 TI - The efficiency of functional mitochondrial replacement in Saccharomyces species has directional character. AB - Optimal interactions among nuclear and mitochondria-coded proteins are required to assemble functional complexes of mitochondrial oxidative phosphorylation. The communication between the nuclear and mitochondrial genomes has been studied by transplacement of mitochondria from related species into mutants devoid of mitochondrial DNA (rho0). Recently we have reported that the mitochondria transferred from Saccharomyces paradoxus restored partially the respiration in Saccharomyces cerevisiae rho0 mutants. Here we present evidence that the S. cerevisiae mitochondria completely salvage from respiration deficiency, not only in conspecific isolates but also in S. paradoxus. The respiratory capacity in less-related species can be recovered exclusively in the presence of S. cerevisiae chromosomes. The efficiency of the re-established oxidative phosphorylation did not rely on the presence of introns in the S. cerevisiae mitochondrial DNA. Our results suggest that, apart from evolutionary distance, the direction of mitochondrial replacement could play a significant role in installing the complete (wild-type-like) interaction between mitochondria and nuclei from different species. PMID- 14554202 TI - Geography and niche occupancy as determinants of yeast biodiversity: the yeast insect-morning glory ecosystem of Kipuka Puaulu, Hawai'i. AB - Biodiversity theory proposes two types of hypotheses to account for the species composition of a given community. The first encompasses geographic and historical factors. For example, local species richness is thought to be affected by area, proximity to large landmasses, dispersal mechanisms, and climatic history, collectively known as biogeography. The second type, termed niche occupancy rules, deals with the intrinsic properties of the species as they affect their interaction with the habitat and with other members of the community. The yeast insect-morning glory ecosystem is a good model to explore biodiversity theory in ascomycetous yeasts. Here we focus on beetles that breed or feed in morning glories and a group of ascomycetous yeasts that are associated exclusively with them. Specifically, we analyse the community found in the vicinity of Kipuka Puaulu, a small patch of disturbed but mature forest situated amidst lava flows on the island of Hawai'i. Major members of the yeast community include Metschnikowia hawaiiensis, Metschnikowia lochheadii, and the related asexual species Candida ipomoeae and Candida kipukae. These species are nearly indistinguishable from one another in terms of nutritional requirements and abilities, although their phylogenetic range is enormous. Their distribution, both global and local, is far from random. As Kipuka Puaulu is an island within an island, the principles of island biogeography may be invoked to explain some aspects of its yeast species composition. M. lochheadii, C. ipomoeae, and the rare species Candida hawaiiana are recent introductions from the American continent and therefore exotic, whereas M. hawaiiensis and C. kipukae might be regarded as endemic, as they are yet to be isolated elsewhere. Vectoring by certain nitidulid beetles explains the long-range dispersal of these species. However, niche occupancy rules may account in part for the local spatial distribution of the yeasts within the island of Hawai'i and within the kipuka itself. We have identified the maximum growth temperature as a potentially critical property of the fundamental niche of these yeasts. PMID- 14554203 TI - Issatchenkia hanoiensis, a new yeast species isolated from frass of the litchi fruit borer Conopomorpha cramerella Snellen. AB - The new ascogenous yeast species Issatchenkia hanoiensis was discovered in the frass of the litchi fruit borer Conopomorpha cramerella Snellen. The yeast forms unconjugated persistent asci containing one to two roughened ascospores. The yeast has a CoQ-7 system, which is typical for the genus Issatchenkia. The closest species to I. hanoiensis as indicated by analysis of the partial ribosomal DNA large-subunit (D1/D2) sequence is the asexual species Candida pseudolambica. The two share 94.2% similarity in the sequenced region. Other species of Issatchenkia were also among the closest relatives of I. hanoiensis, the level of similarity ranging from 89.8% to 94.1%. The type culture is strain HB1.3.13=CBS 9198=NRRL Y-27509. PMID- 14554204 TI - Comparative analyses of Saccharomyces cerevisiae RNAs using Agilent RNA 6000 Nano Assay and agarose gel electrophoresis. AB - Precise quantification and quality characterisation of isolated RNAs are prerequisites for their further exploitation in genome-wide microarrays, Northern blots, cDNA library preparation and others. Our data indicate that RNA analyses using Agilent RNA Nano Assay exhibit several advantages when compared with those performed on ethidium bromide-stained agarose gel electrophoresis or on a spectrophotometer. The RNA Nano Assay makes it possible to estimate RNA concentrations in the range from 1000 ng microl(-1) to 17 ng microl(-1). The presence of impurities including traces of DNA within RNA samples does not influence the concentration measurements. Like agarose gel electrophoresis, RNA Nano Assay allows to analyse RNAs dissolved in formamide and therefore protected against RNase action. Moreover, it allows a clearer distinction of partially degraded samples. The limitation of RNA Nano Assay is the impossibility to detect and to analyse double-stranded RNAs. PMID- 14554206 TI - Differential effects of static and dynamic compression on meniscal cell gene expression. AB - Cells of the meniscus are exposed to a wide range of time-varying mechanical stimuli that may regulate their metabolic activity in vivo. In this study, the biological response of the meniscus to compressive stimuli was evaluated in vitro, using a well-controlled explant culture system. Gene expression for relevant extracellular matrix proteins was quantified using real-time RT-PCR following a 24 h period of applied static (0.1 MPa compressive stress) or dynamic compression (0.08-0.16 MPa). Static and dynamic compression were found to differentially regulate mRNA levels for specific proteins of the extracellular matrix. Decreased mRNA levels were observed for decorin ( approximately 2.1 fold difference) and type II collagen ( approximately 4.0 fold-difference) following 24 h of dynamic compression. Decorin mRNA levels also decreased following static compression ( approximately 4.5 fold-difference), as did mRNA levels for both types I ( approximately 3.3 fold-difference) and II collagen ( approximately 4.0 fold-difference). Following either static or dynamic compression, mRNA levels for aggrecan, biglycan and cytoskeletal proteins were unchanged. It is noteworthy that static compression was associated with a 2.6 fold-increase in mRNA levels for collagenase, or MMP-1, suggesting that the homeostatic balance between collagen biosynthesis and catabolism was altered by the mechanical stimuli. These findings demonstrate that the biosynthetic response of the meniscus to compression is regulated, in part, at the transcriptional level and that transcription of types I and II collagen as well as decorin may be regulated by common mechanical stimuli. PMID- 14554207 TI - cDNA-arrays and real-time quantitative PCR techniques in the investigation of chronic Achilles tendinosis. AB - The aetiology and pathogenesis of chronic painful Achilles tendinosis are unknown. This investigation aimed to use cDNA arrays and real-time quantitative polymerase chain reaction (real-time PCR) technique to study tendinosis and control tissue samples. Five patients (females mean age 57.1+/-4.3 (years+/-SD)) with chronic painful Achilles tendinosis were included. From all patients, one biopsy was taken from the area with tendinosis and one from a clinically normal area (control) of the tendon. The tissue samples were immediately immersed in RNAlater and frozen at -80 degrees C until RNA extraction. Portions of pooled RNA from control and tendinosis sites, respectively, were transcribed to cDNA, radioactively labelled (32P), hybridized to cDNA expression arrays, and exposed to phosphoimager screens over night. Expressions of specific genes, shown to be regulated in the cDNA array analysis, were analyzed in the individual samples using real-time PCR. cDNA arrays showed that gene expressions for matrix metalloproteinase-2 (MMP-2), fibronectin subunit B (FNRB), vascular endothelial growth factor (VEGF), and mitogen-activated protein kinase p38 (MAPKp38) were up regulated, while matrix-metalloproteinase-3 (MMP-3) and decorin were down regulated, in tendinosis tissue compared with control tissue. Using real-time PCR, 4/5 and 3/5 patients showed up-regulation of MMP-2 and FNRB mRNA, respectively. For decorin, VEGF, and MAPKp38, real-time PCR revealed a great variability among patients. Interestingly, the mRNAs for several cytokines and cytokine receptors were not regulated, indicating the absence of an inflammatory process in chronic painful Achilles tendinosis. In conclusion, cDNA-arrays and real-time PCR can be used to study differences in gene expression levels between tendinosis and control tendon tissue. PMID- 14554208 TI - Gastric pentadecapeptide BPC 157 accelerates healing of transected rat Achilles tendon and in vitro stimulates tendocytes growth. AB - In studies intended to improve healing of transected Achilles tendon, effective was a stable gastric pentadecapeptide BPC 157 (GEPPPGKPADDAGLV, M.W. 1419). Currently in clinical trials for inflammatory bowel disease (PLD-116, PL 14736, Pliva), it ameliorates internal and external wound healing. In rats, the right Achilles tendon transected (5 mm proximal to its calcaneal insertion) presents with a large tendon defect between cut ends. Agents (/kg b.w., i.p., once time daily) (BPC 157 (dissolved in saline, with no carrier addition) (10 microg, 10 ng or 10 pg) or saline (5.0 ml)), were firstly applied at 30 min after surgery, the last application at 24 h before autopsy. Achilles functional index (AFI) was assessed once time daily. Biomechanical, microscopical and macroscopical assessment was on day 1, 4, 7, 10 and 14. Controls generally have severely compromised healing. In comparison, pentadecapeptide BPC 157 fully improves recovery: (i) biomechanically, increased load of failure, load of failure per area and Young's modulus of elasticity; (ii) functionally, significantly higher AFI-values; (iii) microscopically, more mononuclears and less granulocytes, superior formation of fibroblasts, reticulin and collagen; (iv) macroscopically, smaller size and depth of tendon defect, and subsequently the reestablishment of full tendon integrity. Likewise, unlike TGF-beta, pentadecapeptide BPC 157, presenting with no effect on the growth of cultured cell of its own, consistently opposed 4-hydroxynonenal (HNE), a negative modulator of the growth. HNE-effect is opposed in both combinations: BPC 157+HNE (HNE growth inhibiting effect reversed into growth stimulation of cultured tendocytes) and HNE+BPC 157(abolished inhibiting activity of the aldehyde), both in the presence of serum and serum deprived conditions. In conclusion, these findings, particularly, Achilles tendon transection fully recovered in rats, peptide stability suitable delivery, usefully favor gastric pentadecapeptide BPC 157 in future Achilles tendon therapy. PMID- 14554209 TI - Shock wave therapy induces neovascularization at the tendon-bone junction. A study in rabbits. AB - Despite the success in clinical application, the exact mechanism of shock wave therapy remains unknown. We hypothesized that shock wave therapy induces the ingrowth of neovascularization and improves blood supply to the tissues. The purpose of this study was to investigate the effect of shock wave therapy on neovascularization at the tendon-bone junction. Fifty New Zealand white rabbits with body weight ranging from 2.5 to 3.5 kg were used in this study. The right limb (the study side) received shock wave therapy to the Achilles tendon near the insertion to bone. The left limb (the control side) received no shock wave therapy. Biopsies of the tendon-bone junction were performed in 0, 1, 4, 8 and 12 weeks. The number of neo-vessels was examined microscopically with hematoxylin eosin stain. Neovascularization was confirmed by the angiogenic markers including vessel endothelial growth factor (VEGF) and endothelial nitric oxide synthase (eNOS) expressions and endothelial cell proliferation determined by proliferating cell nuclear antigen (PCNA) expression examined microscopically with immunohistochemical stains. The results showed that shock wave therapy produced a significantly higher number of neo-vessels and angiogenesis-related markers including eNOS, VEGF and PCNA than the control without shock wave treatment. The eNOS and VEGF began to rise in as early as one week and remained high for 8 weeks, then declined at 12 weeks; whereas the increases of PCNA and neo-vessels began at 4 weeks and persisted for 12 weeks. In conclusion, shock wave therapy induces the ingrowth of neovascularization associated with early release of angiogenesis-related markers at the Achilles tendon-bone junction in rabbits. The neovascularization may play a role to improve blood supply and tissue regeneration at the tendon-bone junction. PMID- 14554210 TI - Bone loss following tendon laceration, repair and passive mobilization. AB - Little is known about the localized changes in bone mass that occur following tendon or ligament injury. Interruption of normal load transfer at the insertion site will presumably lead to a localized loss of bone, although few data exist to support this claim. To test this hypothesis, we transected the canine flexor digitorum profundus (FDP) tendon from its insertion, and either repaired it using a trans-osseous suture technique or left it unrepaired (laceration only). Post operatively, forelimbs in the repair group were cast immobilized except for 10 min of daily passive mobilization rehabilitation, whereas in the laceration only group dogs were allowed full weight bearing. At 5-42 days post-injury, we assessed bone mineral density (BMD) using pQCT and osteoclast surface by histomorphometry. We measured significant bone loss in the distal phalanx after combined FDP tendon laceration, repair, and post-operative passive mobilization, with BMD decreases of 20%, 40%, and 41% at 10, 21, and 42 days (p<0.01). Moreover, we observed that passive mobilization and tendon laceration each contributed independently to the observed bone loss. At 42 days, BMD was reduced by 21% in bones that were not injured but were subjected to the post-operative passive mobilization protocol, while BMD was reduced by 28% in bones subjected to tendon laceration and full weight bearing (p<0.01). In both the passive mobilization and laceration specimens, we counted significantly increased osteoclasts after only 7-10 days, and these increases persisted through 42 days (p<0.05). We conclude that rapid and sustained bone resorption leads to significant bone loss in the 6-week period following flexor tendon injury and repair. This bone loss may impact healing by impeding the restoration of a strong tendon-bone interface. PMID- 14554211 TI - rhBMP-2 injected in a calcium phosphate paste (alpha-BSM) accelerates healing in the rabbit ulnar osteotomy model. AB - This study evaluated the ability of recombinant human bone morphogenetic protein 2 (rhBMP-2) delivered in an injectable calcium phosphate carrier (alpha-BSM) to accelerate healing in a rabbit ulna osteotomy model compared to untreated surgical controls. Healing was assessed by radiography, histology and biomechanics. Bilateral mid-ulnar osteotomies were created in 16 skeletally mature rabbits. One limb in each animal was injected with either 0.1 mg rhBMP 2/alpha-BSM (BMP) (N=8) or buffer/alpha-BSM (BSM) (N=8). Contralateral osteotomies served as untreated surgical controls (SXCT). Gamma scintigraphy showed 75%, 45% and 5% of the initial 125I-rhBMP-2 dose was retained at the osteotomy site at 3 h, 1 week and 3 weeks. The biological activity of rhBMP-2 (alkaline phosphatase activity from bioassay) extracted from alpha-BSM incubated in vitro up to 30 days at 37 degrees C was unchanged. Radiographs demonstrated complete bridging of the BMP limbs at 4 weeks whereas none of the BSM or SXCT limbs were bridged. Post-mortem peripheral quantitative computed tomography determined mineralized callus area was 62% greater in BMP limbs compared to SXCT limbs. Torsional stiffness and strength were 63% and 103% greater in BMP limbs compared to SXCT limbs. There was no difference in torsional properties between BSM and SXCT limbs. Failure occurred outside the osteotomy in four out of seven of the BMP limbs. All BSM and SXCT limbs failed through the osteotomy. Histology showed bony bridging of the osteotomy and no residual carrier in the BMP limbs. BSM and SXCT groups showed less mature calluses composed of primarily fibrocartilaginous tissue and immature bone in the osteotomy gap. These data indicate rhBMP-2 delivered in alpha-BSM accelerated healing in a rabbit ulna osteotomy model compared to BSM and SXCT groups. PMID- 14554212 TI - Cryopreservation of tissue engineered constructs for bone. AB - The large-scale clinical use of tissue engineered constructs will require provisions for its mass availability and accessibility. Therefore, it is imperative to understand the effects of low temperature (-196 degrees C) on the tissue engineered biological system. Initial studies used samples of the osteoblast-like cell line (SaOS-2) adhered to a two-dimensional poly(lactide-co glycolide) thin film (2D-PLAGA) or a three-dimensional poly(lactide-co-glycolide) sintered microsphere matrix (3D-PLAGA) designed for bone tissue engineering. Experimental samples were tested for their ability to maintain cell viability, following low temperature banking for one week, in solutions of the penetrating cryoprotective agents, dimethylsulfoxide (DMSO), ethylene glycol, and glycerol. Results indicated the DMSO solution yielded the greatest percent cell survival for SaOS-2 cells adhered to both the 2D- and 3D-PLAGA scaffolds; therefore, DMSO was used to cryopreserve mineralizing primary rabbit osteoblasts cells adhered to 2D-PLAGA matrices for 35 days. Results indicated retention of the extracellular matrix architecture as no statistically significant difference in the pre- and post-thaw mineralized structures was measured. Percent cell viability of the mineralized constructs following low temperature storage was approximately 50%. These are the first studies to address the issue of preservation techniques for tissue engineered constructs. The ability to successfully cryopreserve mineralized tissue engineered matrices for bone may offer an unlimited and readily available source of bone-like materials for orthopaedic applications. PMID- 14554213 TI - Shear movement at the fracture site delays healing in a diaphyseal fracture model. AB - This study tested the hypothesis that interfragmentary axial movement of transverse diaphyseal osteotomies would result in improved fracture healing compared to interfragmentary shear movement. Ten skeletally mature merino sheep underwent a middiaphyseal osteotomy of the right tibia, stabilized by external fixation with an interfragmentary gap of 3 mm. A custom made external fixator allowed either pure axial (n=5) or pure shear movement (n=5) of 1.5 mm amplitude during locomotion by the animals. The movement of the osteotomy gap was monitored weekly in two sheep by an extensometer temporarily attached to the fixator. After 8 weeks the sheep were killed, and healing of the osteotomies was evaluated by radiography, biomechanical testing, and undecalcified histology. Shear movement considerably delayed the healing of diaphyseal osteotomies. Bridging of the osteotomy fragments occurred in all osteotomies in the axial group (100%), while in the shear group only three osteotomies (60%) were partially bridged. Peripheral callus formation in the shear group was reduced by 36% compared to the axial group (p<0.05). In the axial group bone formation was considerably larger at the peripheral callus and in between the osteotomy gaps but not in the intramedullary area. The larger peripheral callus and excess in bone tissue at the level of the gap resulted in a more than three times larger mechanical rigidity for the axial than for the shear group (p<0.05). In summary, fixation that allows excessive shear movement significantly delayed the healing of diaphyseal osteotomies compared to healing under axial movement of the same magnitude. PMID- 14554214 TI - Noninvasive fatigue fracture model of the rat ulna. AB - Fatigue damage occurs in response to repeated cyclic loading and has been observed in situ in cortical bone of humans and other animals. When microcracks accumulate and coalesce, failure ensues and is referred to as fatigue fracture. Experimental study of fatigue fracture healing is inherently difficult due to the lack of noninvasive models. In this study, we hypothesized that repeated cyclic loading of the rat ulna results in a fatigue fracture. The aim of the study was to develop a noninvasive long bone fatigue fracture model that induces failure through accumulation and coalescence of microdamage and replicates the morphology of a clinical fracture. Using modified end-load bending, right ulnae of adult Sprague-Dawley rats were cyclically loaded in vivo to fatigue failure based on increased bone compliance, which reflects changes in bone stiffness due to microdamage. Preterminal tracer studies with 0.8% Procion Red solution were conducted according to protocols described previously to evaluate perfusion of the vasculature as well as the lacunocanalicular system at different time points during healing. Eighteen of the 20 animals loaded sustained a fatigue fracture of the medial ulna, i.e. through the compressive cortex. In all cases, the fracture was closed and non-displaced. No disruption to the periosteum or intramedullary vasculature was observed. The loading regime did not produce soft tissue trauma; in addition, no haematoma was observed in association with application of load. Healing proceeded via proliferative woven bone formation, followed by consolidation within 42 days postfracture. In sum, a noninvasive long bone fatigue fracture model was developed that lends itself for the study of internal remodeling of periosteal woven bone during fracture healing and has obvious applications for the study of fatigue fracture etiology. PMID- 14554215 TI - Bone architecture and disc degeneration in the lumbar spine of mice lacking GDF-8 (myostatin). AB - GDF-8, also known as myostatin, is a member of the transforming growth factor beta superfamily of secreted growth and differentiation factors that is expressed in vertebrate skeletal muscle. Myostatin functions as a negative regulator of skeletal muscle growth and myostatin null mice show a doubling of muscle mass compared to normal mice. We describe here morphology of the lumbar spine in myostatin knockout (Mstn(-/-)) mice using histological and densitometric techniques. The Mstn(-/-) mice examined in this study weigh approximately 10% more than controls (p<0.001) but the iliopsoas muscle is over 50% larger in the knockout mice than in wild-type mice (p<0.001). Peripheral quantitative computed tomography (pQCT) data from the fifth lumbar vertebra show that mice lacking myostatin have approximately 50% greater trabecular bone mineral density (p=0.001) and significantly greater cortical bone mineral content than normal mice. Toluidine blue staining of the intervertebral disc between L4-L5 reveals loss of proteoglycan staining in the hyaline end plates and inner annulus fibrosus of the knockout mice. Loss of cartilage staining in the caudal end plate of L4 is due to ossification of the end plate in the myostatin-deficient animals. Results from this study suggest that increased muscle mass in mice lacking myostatin is associated with increased bone mass as well as degenerative changes in the intervertebral disc. PMID- 14554216 TI - The effects of sex and estrogen therapy on bone ingrowth into porous coated implant. AB - Amounts of bone ingrowth into porous cobalt-chromium plugs were compared between male and female dogs, and among sham-operated and ovariectomized female dogs, with or without estrogen treatment, to investigate the effect of gender and estrogen therapy on biologic fixation. Each group consisted of eight skeletally mature dogs. Plugs were implanted bilaterally in the distal femur at 6 months after ovariectomy or sham operation. Estrogen treatment group received estradiol 20 microg/kg/day subcutaneous injection. Three months after implantation, histological examination showed significantly more bone ingrowth in areas with cortical bone contact than in areas with cancellous bone contact (P<0.001 for all groups). Bone ingrowth was essentially the same in male and female control dogs. Ovariectomized dogs showed less overall bone ingrowth than male and female controls (P=0.007). Bone ingrowth in areas with cortical bone contact did not decrease significantly, whereas bone ingrowth in areas with cancellous bone contact was significantly impaired (P<0.001) in ovariectomized dogs compared with female controls. Short-term, high-dose estradiol treatment did not increase bone ingrowth volume fraction. Mechanical tests did not show any statistical differences among groups. CONCLUSION: Type of bone contact is the key factor affecting the amount and pattern of bone ingrowth into the porous surface. Ovariectomy results in decreased bone ingrowth in areas with cancellous bone contact, but does not compromise bone ingrowth in areas with cortical bone contact. Short-term, high-dose estradiol treatment does not enhance bone ingrowth into the porous surface. Extensively coated or full-coated porous prostheses are recommended to achieve enough cortical bone contact and ingrowth for post menopausal patients. PMID- 14554217 TI - Mitogen-activated protein (MAP) kinases mediate PMMA-induction of osteoclasts. AB - Inflammatory osteolysis induced by implant-derived wear debris is associated with infiltration of various cell-types to the implant-bone interface leading to abundant secretion of pro-inflammatory cytokines and activation of proteinases that together lead to propagation of the localized inflammatory response and periprosthetic bone erosion. Tumor necrosis factor family members are considered to be direct mediators of inflammation and osteolysis. These cytokines exert their osteoclastic effects via activation of the transcription factor NF-kappaB and certain MAP kinases, including c-Jun, Erks and p38, all known to be essential for the development of osteoclasts. We have recently documented that the osteoclastogenic cytokines TNF and RANKL play a pivotal role in the development of inflammatory osteolysis. We have also found that PMMA particles stimulate osteoclastogenesis, at least in part, by induction of RANKL, TNF, and by activation of the transcription factor NF-kappaB. More importantly, our data indicate that inhibitors of the osteoclastogenic factors, TNF and RANKL abrogate particle-induced osteoclastogenesis. In the current study, we investigated if PMMA particles activate MAP kinases, and the potential role of these kinases as mediators of osteolysis. Using kinase assays, we show that in osteoclast precursors, PMMA particles markedly and rapidly activate p38 and ERK MAP kinases. This activation was specific, evident by complete blockade with specific inhibitory compounds. Similarly, we show that PMMA particles activate the JNK pathway, which is known to be involved in inflammatory and osteoclastogenic events. We also show that p38 MAP kinase regulates PMMA-activation of NF-kappaB, thus providing a possible mechanism for particle action in osteoclast precursors. Finally, we provide evidence that specific inhibitors of MAP kinases are capable of inhibiting PMMA-stimulated osteoclastogenesis. These data provide evidence that MAP kinases are potent mediators of particle-induced osteoclastogenesis. PMID- 14554219 TI - Detection of chimerism following vascularized bone allotransplantation by polymerase chain reaction using a Y-chromosome specific primer. AB - Chimerism following allogeneic organ transplantation is a phenomenon known to occur and be associated with development of immunologic tolerance in allotransplantation. However, little is known about graft cell migration following vascularized bone allografting. In this study, chimerism was assessed following vascularized tibia transplantation from male DA or PVG donors to female PVG rat recipients using a semi-quantitative polymerase chain reaction for the Y chromosome. FK-506 (Tacrolimus) was administered after transplantation for immunosuppression. All immunosuppresssed PVG rat recipients of PVG bone grafts showed a high level of chimerism (1%) in the thymus, spleen, liver and cervical lymph nodes at 18 weeks post-transplant. Donor cells were also detected in the contralateral tibia and humerus. In non-immunosuppressed PVG rat recipients of DA bone grafts, donor cells were detected in the spleen in three of five rats within 2 weeks post-transplant. In these animals the bone grafts were severely rejected. In immunosuppressed PVG rat recipients of DA bone grafts, two of five, four of eight and eight of 10 rats showed low level chimerism (0.1%) in peripheral blood at 1, 12, and 18 weeks post-transplant. Six rats showed a high level of chimerism in the spleen and thymus. Histological studies revealed no rejection findings through 18 weeks post-transplant. Our results indicate that chimerism, or the presence of graft cells in host tissue, may occur in the face of acute rejection and be demonstrable following vascularized isograft and allograft living bone transplantation when chronic immunosuppression is maintained. Graft vascular patency during the short-term likely allows cellular migration, even in the face of acute rejection. Long-term survival and proliferation of graft marrow elements in host tissue may be possible with adequate immunosuppression. PMID- 14554218 TI - Use of volumetric computerized tomography as a primary outcome measure to evaluate drug efficacy in the prevention of peri-prosthetic osteolysis: a 1-year clinical pilot of etanercept vs. placebo. AB - Although total hip replacement (THR) is amongst the most successful and beneficial medical procedures to date, long-term outcomes continue to suffer from aseptic loosening secondary to peri-prosthetic osteolysis. Extensive research over the last two decades has elucidated a central mechanism for osteolysis in which wear debris generated from the implant stimulates inflammatory cells to promote osteoclastogenesis and bone resorption. The cytokine tumor necrosis factor alpha (TNFalpha) has been demonstrated to be central to this process and is considered to be a leading target for intervention. Unfortunately, even though FDA approved TNF antagonists are available (etanercept), currently there are no reliable outcome measures that can be used to evaluate the efficacy of a drug to prevent peri-prosthetic osteolysis. To the end of developing an effective outcome measure, we evaluated the progression of lesion size in 20 patients with established peri-acetabular osteolysis (mean=29.99 cm(3), range=2.9-92.7 cm(3)) of an uncemented primary THR over 1-year, using a novel volumetric computer tomography (3D-CT) technique. We also evaluated polyethylene wear, urine N telopeptides and functional assessments (WOMAC, SF-36 and Harris Hip Score) for comparison. At the time of entry into the study baseline CT scans were obtained and the patients were randomized to etanercept (25 mg s.q., twice/week) and placebo in a double-blinded fashion. CT scans, urine and functional assessments were also obtained at 6 and 12 months. No serious adverse drug related events were reported, but one patient had to have revision surgery before completion of the study due to aseptic loosening. No remarkable differences between the groups were observed. However, the study was not powered to see significant drug effects. 3D-CT data from the 19 patients was used to determine the mean increase in lesion size over 48 weeks, which was 3.19 cm(3) (p<0.0013). Analysis of the urine N-telopeptides and functional assessment data failed to identify a significant correlation with wear or osteolysis. In conclusion, volumetric CT was able to measure progression of osteolysis over the course of a year, thus providing a technology that could be used in therapeutic trials. Using the data from this pilot we provide a model power calculation for such a trial. PMID- 14554220 TI - Inhibition of MMP-1 expression by antisense RNA decreases invasiveness of human chondrosarcoma. AB - We previously reported that an elevated level of matrix metalloproteinase-1 (MMP 1) gene expression in patients with chondrosarcoma has a strong statistical correlation with recurrence and in vitro invasion. In the present study, we used an antisense RNA strategy for MMP-1 inhibition to determine if this would affect the invasive characteristics of the cells. We transfected a human chondrosarcoma cell line with a retroviral plasmid expressing a 770 bp genomic fragment of the human MMP-1 gene in the sense or antisense orientation. The results show that cells transfected with the MMP-1 antisense fragment had a significant decrease in both MMP-1 protein and enzyme activity (p<0.05) as compared to cells transfected with an empty plasmid or the parental cells. Cells transfected with the MMP-1 antisense fragment demonstrated a significant decrease in their ability to invade the collagen I barrier (p<0.05). The gene expression for MMP-8 and MMP-13 were unaffected in cells transfected with the MMP-1 antisense fragment, MMP-1 sense fragment, or empty plasmid. These results support the hypothesis that MMP-1 facilitates tumor cell egress from chondrosarcoma tissue and demonstrate the potential of MMP-1 as a promising target for a novel biologic therapy in chondrosarcoma. PMID- 14554221 TI - The C-terminal domain of focal adhesion kinase reduces the tumor cell invasiveness in chondrosarcoma cell lines. AB - Human chondrosarcoma is a malignancy that has no effective systemic therapy, making the interruption of the metastatic cascade critical to enhance patient survival. The processes of local invasion and metastases share similar mechanisms at a cellular level. Focal adhesion kinase (FAK) has been implicated in local invasion of malignant tumor cells. In the current manuscript we examine the effect of FAK inhibition on cell attachment to extracellular matrix (ECM) and in vitro invasion. Bovine articular chondrocytes and two human chondrosarcoma cell lines were utilized to examine FAK activity in tumor cell invasiveness. Endogenous FAK activity was inhibited by adenoviral transfection with the C terminal domain of FAK. This inhibition resulted from decreased FAK phosphorylation, while FAK expression remained unchanged. Inhibition of FAK phosphorylation and hence its activity lead to decreased cell adhesion to Type II collagen and decreased cell invasiveness. These effects were not due to changes in integrin expression, indicating that the inhibition was the result of disruption of outside: in signaling. There are three important aspects to these results. The first is that interruption of transmembrane signaling can affect cell attachment. The second is that in chondrosarcoma, cell differentiation correlates with FAK expression and metastatic potential. Thirdly, that cell invasiveness correlates with FAK activity and implies a mechanistic role for this molecular complex in local invasion and metastasis. PMID- 14554222 TI - Fibronectin metabolism of cartilage explants in response to the frequency of intermittent loading. AB - Chondrocytes within articular cartilage experience complete unloading between loading cycles and in so doing utilize mechanical signals to regulate their own metabolic activities. A strongly elevated fibronectin content is an early feature in osteoarthritis and appears to be related to increases in both the synthesis and retention of this glycoprotein. The objectives of this study were to investigate systematically whether the frequency of intermittently applied cyclic mechanical loading of cartilage explants alters the biosynthesis and retention of fibronectin, and to assess whether it is possible to induce in vitro osteoarthritic-like changes of this metabolic parameter by mechanical means over a period of 6 days. Cartilage plugs consisting of viability-checked chondrocytes were exposed to sinusoidal cyclic compressive pressure alterations of 0.1, 0.5 or 1.0 Hz frequency with a peak stress of 0.5 MPa for a period of 5, 10 or 20 s, followed by an unloading period of 10, 100 or 1000 s, and compared to unloaded reference plugs from the same joint and topographic origin. The incorporation of radioactive precursor into fibronectin during the last 18 h, the content of fibronectin, and the viability of chondrocytes were determined. Our data revealed that (a) the fibronectin synthesis was selectively, but non-linearly affected by the frequency of intermittent loads applied (as defined by the frequency of the applied force, the duration of the loading cycle and the duration of the force free period between each loading cycle), and that (b) the retention of endogenous fibronectin and proteins within loaded cartilage explants is strongly elevated. These data support our hypothesis that the mechanical factor "frequency of intermittent loading" seems to be the crucial mechanical parameter controlling the metabolism of chondrocytes. The effect of the frequency of intermittent loading cannot be described by a simple statistical correlation, so that no specific predictions are possible. However, our results imply that distinct loading protocols have been established that can induce alterations of the fibronectin metabolism similar to those observed in human and animal osteoarthritis. PMID- 14554224 TI - Subject-specific finite element analysis of the human medial collateral ligament during valgus knee loading. AB - The objectives of this study were (1) to develop subject-specific experimental and finite element (FE) techniques to study the three-dimensional stress-strain behavior of ligaments, with application to the human medial collateral ligament (MCL), and (2) to determine the importance of subject-specific material properties and initial (in situ) strain distribution for prediction of the strain distribution in the MCL under valgus loading. Eight male knees were subjected to varus-valgus loading at flexion angles of 0 degrees, 30 degrees, and 60 degrees. Three-dimensional joint kinematics and MCL strains were recorded during kinematic testing. Following testing, the MCL of each knee was removed to allow measurement of the in situ strain distribution and to perform material testing. A FE model of the femur-MCL-tibia complex was constructed for each knee to simulate valgus loading at each flexion angle, using subject-specific bone and ligament geometry, material properties, and joint kinematics. A transversely isotropic hyperelastic material model was used to represent the MCL. The MCL in situ strain distribution at full extension was used to apply in situ strain to each MCL FE model. FE predicted MCL strains during valgus loading were compared to experimental measurements using regression analysis. The subject-specific FE predictions of strain correlated reasonably well with experimentally measured MCL strains (R(2)=0.83, 0.72, and 0.66 at 0 degrees, 30 degrees, and 60 degrees, respectively). Despite large inter-subject variation in MCL material properties, MCL strain distributions predicted by individual FE models that used average MCL material properties were strongly correlated with subject-specific FE strain predictions (R(2)=0.99 at all flexion angles). However, predictions by FE models that used average in situ strain distributions yielded relatively poor correlations with subject-specific FE predictions (R(2)=0.44, 0.35, and 0.33 at flexion angles of 0 degrees, 30 degrees, and 60 degrees, respectively). The strain distribution within the MCL was nonuniform and changed with flexion angle. The highest MCL strains occurred at full extension in the posterior region of the MCL proximal to the joint line during valgus loading, suggesting this region may be most vulnerable to injury under these loading conditions. This work demonstrates that subject-specific FE models can predict the complex, nonuniform strain fields that occur in ligaments due to external loading of the joint. PMID- 14554223 TI - Chondrogenic differentiation of mesenchymal progenitor cells encapsulated in ultrahigh-viscosity alginate. AB - One major problem of current cartilage repair techniques is that three dimensional encapsulated mesenchymal progenitor cells frequently differentiate into hypertrophic cells that express type X collagen and osteogenic marker genes. Studies on wild-type cells of murine mesenchymal C3H10T1/2 progenitor cells as well as on cells transfected with cDNA encoding for bone morphogenetic protein (BMP)-2 or -4 in alginate revealed that the formation of markers for osteogenesis and chondrogenic hypertrophy apparently depended on the BMP-transfection. Cells were encapsulated in ultrahigh-viscosity, clinical grade alginate and differentiation was studied over a period of 17 days. Consistent with results published previously staining with haematoxylin-eosin or Alcian blue, immunohistochemical analysis, and quantitative RT-PCR confirmed the expression of chondrogenic markers (chondroitin-4- and -6-sulfate as well as type II collagen). Production of chondrogenic markers was particularly high in BMP-4 transfected cells. Hypertrophic chondrogenesis did not occur in BMP-4 transfected cells, as revealed by measurement of type X collagen, but could be demonstrated for wild type cells and to some extent for BMP-2 transfected cells. The osteogenic markers, type I collagen, alkaline phosphatase, and Cbfa1 were upregulated in all cell lines even though the levels and the time of upregulation differed significantly. In any case, the markers were less and only very shortly expressed in BMP-4 transfected cells as revealed quantitatively by real time RT-PCR. Thus, the in vitro results suggested that BMP-4 is a very promising candidate for suppressing chondrogenic hypertrophy, while simultaneously enhancing the production of chondrogenic components. PMID- 14554225 TI - A quantitative analysis of valgus torque on the ACL: a human cadaveric study. AB - The loads needed to elicit a positive pivot shift test in a knee with an anterior cruciate ligament (ACL) rupture have not been quantified. The coupled anterior tibial translation (ATT), coupled internal tibial rotation (ITR), and the in situ force in the ACL in response to a valgus torque, an inherent component of the pivot shift test, were measured in 10 human cadaveric knee specimens. Using a robotic/universal force-moment sensor testing system, valgus torques ranging from 0.0 to 10.0 Nm were applied in nine increments on the intact and ACL-deficient knee in flexion ranging from 0 degrees to 90 degrees. At 15 degrees of knee flexion, the coupled ATT and ITR were significantly increased in the ACL deficient knee when compared to the intact knee. Coupled ATT increased a maximum of 291% (6.7 mm, p<0.05), while coupled ITR increased a maximum of 85% (5.1 degrees, p<0.05). At 30 degrees, the increases in coupled ATT and ITR were significant at valgus loads of 3.3 Nm and greater with a maximum increase in coupled ATT of 137% (6.3 mm, p<0.05) and a maximum increase in coupled ITR of 38% (3.6 degrees, p<0.05). At 45 degrees, coupled ATT increased significantly (maximum of 69%, 4.4 mm, p<0.05), but only at torques > or =6.7 Nm. The in situ force in the ACL was less than 20 N for all flexion angles when a torque between 3.3 and 5.0 Nm was applied. Low valgus torque elicited tibial subluxation in the ACL-deficient knee with low in situ ACL forces, similar to a positive pivot shift test. Thus, application of a valgus torque may be suitable to evaluate ACL deficient and ACL-reconstructed knees, since subluxation can be achieved with minimal harm to the ACL graft. This work is important in understanding one load component needed for the pivot shift examination; further studies quantifying other load components are essential for better comprehension of the in vivo pivot shift examination. PMID- 14554227 TI - Correlation of healing capacity with vascular response in the anterior cruciate and medial collateral ligaments of the rabbit. AB - In clinical terms, functional recovery after anterior cruciate ligament (ACL) injury is generally poorer than after medial collateral ligament (MCL) injury. In experimental studies of injury, the early phases of ligament healing require an augmented blood supply. We hypothesized that the differences in healing properties of the ACL and MCL would be reflected in the magnitude of their vascular responses to partial injury. This study is the first to quantify and define the time course of changes in blood flow and vascular volume following hemisection of the rabbit ACL and MCL. Adult female rabbits were assigned to control, sham operation, ACL hemisection or MCL hemisection groups. Standardized ACL or MCL injuries were surgically induced. About 2, 6 or 16 weeks later, blood flow and vascular volume of the ACL and MCL were measured. The MCL of the rabbit responded to hemisection with a large significant increase in blood flow and a substantial angiogenic response associated with inflammation and scar formation. During subsequent matrix remodelling, blood flow and vascular volume returned towards control values. In contrast, the ACL showed only a 2-fold increase in vascular volume, no increase in blood flow and atrophied after hemisection. The superior capacity of the MCL to increase its blood supply through angiogenesis and increased flow is essential for ligament healing to occur, and may be the major difference in healing potential between the ACL and MCL. PMID- 14554226 TI - Effects of transcutaneous topical injection of oxygen on vascular endothelial growth factor gene into the healing ligament in rats. AB - The effects of intermittent exposure to oxygen injection on an experimentally induced ligament tear were studied in the right hind limb of 17 male Sprague Dawley rats. Two rats were used for monitoring the partial oxygen pressure (pO(2)) of subcutaneous tissue and 15 rats were divided into the following three groups of 5 after an experimentally induced ligament tear: Group A, control group; Group B, injection of 0.5 ml hyaluronan to the wound transcutaneously; Group C, injection of 0.5 ml hyaluronan mixed with haemoglobin and oxygen (n=5). At 7 days post-ligament injury, we compared the ligaments of the three treatment groups for gross appearance, histology and expression of vascular endothelial growth factor (VEGF) mRNA by RT-PCR. Our results indicate that the pO(2) was immediately elevated to 334.6 mmHg by topical oxygen injection and this method was effective in promoting vessel formation in comparison to the control group (p<0.01). However, the expression of VEGF mRNA in the topical oxygen injection group (Group C) was lower than that in control group (p<0.05). Our results suggest that oxygen is able to accelerate vessel formation in spite of its effect of decreasing VEGF mRNA. Our method of using topical injection proved to be useful in healing the ligament and the wound. PMID- 14554228 TI - The healing medial collateral ligament following a combined anterior cruciate and medial collateral ligament injury--a biomechanical study in a goat model. AB - The ideal treatment of a combined anterior cruciate ligament (ACL) and medial collateral ligament (MCL) injury to the knee is still debated. In particular, the question of whether reconstruction of the ACL can provide the knee with sufficient multidirectional stability to allow for effective MCL healing needs to be better elucidated. Therefore, the first objective of this study was to quantify the changes in the function of goat knees between time-zero and 6 weeks following a combined ACL/MCL injury treated with ACL reconstruction. Using a robotic/universal force-moment sensor testing system, the kinematics of the knee and in situ forces in the ACL/ACL graft as well as in the sham-operated and healing MCL were evaluated in response to (1) a 67 N anterior-posterior (A-P) tibial load and (2) a 5 Nm varus-valgus (V-V) moment. The second objective was to evaluate the structural properties of the healing femur-MCL-tibia complex (FMTC) and the mechanical properties of the healing MCL at 6 weeks under uniaxial tension. In response to the 67 N A-P tibial load, the A-P translations for the experimental knee increased by as much as 4.5 times from time-zero to 6 weeks (p<0.05). Correspondingly, the in situ forces in the ACL graft decreased by as much as 45% (p<0.05). There was no measurable changes of the in situ force in the healing MCL. In response to a 5 Nm V-V moment, V-V rotations were twice as much as controls, but similar for both time periods. From time-zero to 6 weeks, the in situ forces in the ACL graft dropped by over 71% (p<0.05), while the in situ force in the healing MCL was as much as 35+/-19 N. In terms of the structural properties of the healing FMTC, the stiffness and ultimate load values at 6 weeks reached 53% and 29% of sham-operated contralateral controls, respectively (p<0.05). For the mechanical properties of the healing MCL substance, the values for tangent modulus and tensile strength were only 13% and 10% of sham-operated controls, respectively (p<0.05). These results suggest that the ACL graft stabilized the knee initially, but became loose over time. As a result, the healing MCL may have been required to take on excessive loads and was unable to heal sufficiently as compared to an isolated MCL injury. PMID- 14554229 TI - Specificity of muscle action after anterior cruciate ligament injury. AB - Neuromuscular control is believed to be a critical factor in dynamic knee stability. The purpose of this study was to evaluate voluntary muscle control in anterior cruciate ligament deficient (ACL-D) and uninjured people. Twenty athletes of similar age participated in this study. Subjects performed a target matching protocol that required them to produce isometric moments about the knee with fine control in flexion, extension, varus, and valgus (i.e., loads were generated in the plane perpendicular to the long axis of the shank). Electromyographic data were collected from 10 muscles that span the knee. A specificity index was calculated for each muscle to describe how fine-tuned (specific) its muscle activity pattern was with respect to its principal direction of action in the load plane. Diminished specificity of muscle action was observed in 8 of 10 muscles in the ACL-D subjects' involved knees when compared with the activity patterns from their uninvolved knees and those from the uninjured subjects' knees. The vastus lateralis muscle was especially affected. Increased and more global co-contraction was also observed in the ACL-D limbs. The alterations in muscle firing patterns observed in this study are consistent with diminished neuromuscular control. PMID- 14554230 TI - Vascular endothelial growth factor 121 and 165 in the subacromial bursa are involved in shoulder joint contracture in type II diabetics with rotator cuff disease. AB - Vascular endothelial growth factor (VEGF) is a glycoprotein that plays an important role in neovascularization and increases vascular permeability. We reported that VEGF is involved in motion pain of patients with rotator cuff disease by causing synovial proliferation in the subacromial bursa (SAB). The present study investigates whether VEGF is also involved in the development of shoulder contracture in diabetics with rotator cuff disease. We examined 67 patients with rotator cuff disease, including 36 with complete cuff tears, 20 with incomplete tears, and 11 without apparent tears (subacromial bursitis). The patients were into groups according to the presence or absence of diabetes (14 type II diabetics and 53 non-diabetics). Specimens of the synovium of the SAB were obtained from all patients during surgery. Expression of the VEGF gene in the synovium of the subacromial bursa was evaluated by using the reverse transcriptase polymerase chain reaction. The VEGF protein was localized by immunohistochemistry, and the number of vessels was evaluated based on CD34 immunoreactivity. The results showed that VEGF mRNA was expressed in significantly more diabetics (100%, 14/14) than in non-diabetics (70%, 37/53) (P=0.0159, Fisher's test). Investigation of VEGF isoform expression revealed VEGF121 in all 14 diabetics and in 37 of the 53 non-diabetics, VEGF165 in 12 of the 14 diabetics and in 21 of the 53 non-diabetics, and VEGF189 in 1 of the 14 diabetics and in 2 of the 53 non-diabetics. No VEGF206 was expressed in either group. VEGF protein was localized in both vascular endothelial cells and synovial lining cells. The mean number of VEGF-positive vessels and the vessel area were also significantly greater in the diabetics (p<0.015, Mann-Whitney U test). Synovial proliferation and shoulder joint contracture were more common in the diabetics (P=0.0329 and P=0.073, respectively; Fisher's test). The mean preoperative range of shoulder motion significantly differed in terms of elevation between two groups: 103.8 degrees in diabetics and 124.9 degrees in no diabetics (p=0.0039 Mann-Whitney U test). In contrast, external rotation did not significantly differ: 44 degrees in diabetics and 49 degrees in non-diabetics (p=0.4957, Mann-Whitney U test). These results suggest that VEGF121 and VEGF165 expression in the SAB is responsible for the development of shoulder joint contracture, especially in elevation, among type II diabetic patients with rotator cuff disease. PMID- 14554231 TI - Acetabular blood flow during Bernese periacetabular osteotomy: an intraoperative study using laser Doppler flowmetry. AB - BACKGROUND: The blood flow to the acetabular fragment is of some concern in juxtaarticular pelvic osteotomies used for the treatment of hip dysplasia. No direct measurements have determined the effect of the Bernese periacetabular osteotomy (PAO) on acetabular perfusion. METHODS: Acetabular perfusion was measured by means of laser Doppler flowmetry in 10 patients undergoing a PAO for symptomatic acetabular dysplasia. During the surgical procedure, the intraosseous high energy laser Doppler reliably depicts dynamic changes of small vessel blood flow. Measurements were performed after defined surgical steps to obtain sequential information on the blood perfusion of the acetabular fragment. RESULTS: After complete separation of the acetabular fragment, nine out of 10 patients had pulsatile signals, but the blood flow (BF) significantly decreased by 77%. Corrective positioning of the fragment induced no further drop of the BF signal but a loss of pulsatility in six patients. After a recovery period of about 30 min following preliminary fixation of the fragment, reestablishment of the pulsatile signal and an increase of the BF signal was seen. At termination of the surgical procedure, five out of eight patients, who could be followed throughout the whole procedure, showed a clear pulsatile signal in the supraacetabular area. Bleeding of the supraacetabular cancellous surface could be observed in all acetabula. CONCLUSION: Despite careful preservation of soft tissues during the surgical procedure, a significant reduction of the blood flow in the supraacetabular region has been observed. Nevertheless, a pulsatile signal in more than 60% of the fragments after fragment correction and an increasing signal during the recovery period showed ongoing blood perfusion indicating reversible changes in the measured supraacetabular area. All osteotomies healed within eight weeks without showing signs of necrosis during a minimum follow up of 1 year. PMID- 14554232 TI - Quantifying thumb rotation during circumduction utilizing a video technique. AB - Thumb rotation is an essential component of circumduction in order to achieve pulp to pulp contact. In order to evaluate opponensplasty, a device was developed to quantify thumb rotation utilizing a special jig to hold the hand and video analysis. Twenty-seven healthy volunteers (12 female and 15 male) were tested to obtain normative data. Thumb rotation was measured as the difference in angle between the thumb in abduction and extension and maximal circumduction. Rotation angles varied from 70 degrees to 110 degrees. No gender or right/left differences could be detected. We concluded that comparing rotation of the non-injured hand to the injured hand in evaluating opponensplasties can be used as an accurate measurement of thumb circumduction and opposition. PMID- 14554233 TI - Do oncologists have a role in emergency medicine? PMID- 14554234 TI - Amifostine and chemoradiation therapy: ASCO responds. PMID- 14554235 TI - Close relatives or distant cousins? PMID- 14554236 TI - Infections in patients with cancer undergoing chemotherapy: aetiology, prevention, and treatment. AB - Patients with cancer who are undergoing chemotherapy are highly susceptible, especially if neutropenic, to almost any type of bacterial or fungal infection. These infections cause substantial morbidity and mortality. Prophylactic use of antibiotics should be avoided, however, since this practice is associated with a risk of emergence of resistant bacteria and it does not lower the risk of death. However, chemoprophylaxis has a role for candidal fungal infections. Because infection in a neutropenic host can be rapidly fatal if not treated, the empirical administration of broad-spectrum intravenous antibiotics is generally indicated for these patients, and the local frequencies, susceptibility, and resistance patterns of various pathogens must be taken into account. Once therapy has been initiated, changes in antibiotic regimens during the first 5 days are useless unless the patient's clinical condition deteriorates substantially. The treatment of invasive fungal infections is particularly difficult. Many unsolved questions remain, and studies are proposed here that may shed light on these issues. PMID- 14554237 TI - Novel FLT3 point mutation in acute myeloid leukaemia. PMID- 14554238 TI - Inhibitors of cyclo-oxygenase 2: a new class of anticancer agents? AB - Experimental studies have shown that cyclo-oxygenase 2 (COX2) is involved in tumour development and progression. Selective inhibitors of COX2 (coxibs) block tumour growth through many mechanisms, especially by antiangiogenic and proapoptotic effects. In experimental models, coxibs potentiate the activity of cytotoxic agents, hormones, and radiotherapy. Large clinical studies have shown chemopreventive activity of coxibs in colorectal cancer. The findings of preclinical studies coupled with the overexpression of COX2 observed in advanced human tumours are the basis for new therapeutic anticancer strategies based on combinations of coxibs with other anticancer treatment modalities. Early clinical studies have documented the feasibility, good tolerability, and promising activity of coxibs combined with chemotherapy in patients with advanced colorectal and non-small-cell lung cancers. Here, we describe the recent findings on the antitumour effects of coxibs with particular focus on the opportunities that have emerged for treatment of cancer. PMID- 14554239 TI - Cancer-related fatigue: can exercise physiology assist oncologists? AB - Most patients with cancer experience fatigue, a severe activity-limiting symptom with a multifactorial origin. To avoid cancer-related fatigue, patients are frequently advised to seek periods of rest and to reduce their amount of physical activity. This advice is reminiscent of that formerly given to patients with heart disease. However, such recommendations can paradoxically compound symptoms of fatigue, since sedentary habits induce muscle catabolism and thus cause a further decrease in functional capacity. By contrast, there is scientific evidence that an exercise programme of low to moderate intensity can substantially reduce cancer-related fatigue and improve the quality of life of these patients. Current knowledge, combined with findings soon to be published, could launch new opportunities for patients with cancer. In this new century, exercise physiology could soon prove to be very useful for oncologists. PMID- 14554240 TI - Good practice and quality assurance in surgical oncology. AB - The Halstedian era of radical surgical extirpation for solid tumours dominated the first half of the 20th century. But as understanding of cancer biology increased, a paradigm shift occurred which moved the focus away from extensive surgery towards less radical procedures. Although surgery is a recognised factor in local disease control, prognosis is now believed to be predetermined at the time of diagnosis by the presence of micrometastatic deposits. Modern cancer management consists of more skilled and conservative surgery to remove the primary tumour; adjuvant therapies are also given before and after the operation to target the subclinical metastatic deposits. The most important components of high-quality care in surgical oncology are: sound clinical judgment, surgical skill, and multidisciplinary care. These prerequisites are best achieved by specialisation, but high operative volume is not essential for excision of many types of tumour. Quality assurance using several readily available tools can ensure that the process of care from presentation to outcome is constantly improved and that institutional variations in number of cases and quality of care are monitored. PMID- 14554241 TI - The role of nurse-led care in cancer management. AB - Extending nurses' roles and responsibilities so that they may take on some functions of doctors is widely advocated to assist with shortages of medical staff, improve service provision, and to reduce costs. In cancer care in particular, use of specialist nurses to help meet targets for faster diagnosis and treatment is seen as essential. However, there has been little detailed investigation of the consequences, effectiveness, or acceptability of doctor nurse substitution across health care, or more specifically in cancer services. In this article, I review the evidence for nurse-led care in cancer. PMID- 14554242 TI - Laparoscopic prophylactic surgery for HNPCC gene mutation carrier: has the time come? PMID- 14554243 TI - Chromosome arm 8p and cancer: a fragile hypothesis. AB - Chromosome arm 8p is one of the most frequently altered regions in human cancers. Several potential oncogenes and tumour suppressor genes have been identified but further investigations are needed to confirm which are bona fide oncogenic targets. In cancer cells, chromosome breaks may occur at fragile sites throughout the genome. Some fragile sites lie within genes that may have a role in cancer; the best example is FHIT at 3p14, which contains the fragile site FRA3B. We have found that chromosome breaks disrupt the NRG1 gene at 8p12 in breast and pancreatic cancers. We hypothesise that alteration of the NRG1 gene could occur through breakage at a non-common fragile site. PMID- 14554246 TI - Structural requirements for TLR4-mediated LPS signalling: a biological role for LPS modifications. AB - Cells of the mucosal lining are the first to encounter invading bacteria during infection, and as such, they have developed numerous ways of detecting microbial intruders. Recently, we showed that epithelial cells recognize lipopolysaccharide (LPS) through the CD14-Toll-like receptor (TLR)-4 complex. Here, we identify the substructures of LPS that are recognized by the TLR4 receptor complex. In contrast to lipid A, the O-antigen does not mediate an inflammatory response; rather it interferes with the lipid A recognition. An Escherichia coli strain genetically modified to express penta-acylated lipid A not only showed reduced immunogenicity, but was also found to inhibit pro-inflammatory signalling induced by wild-type E. coli (hexa-acylated lipid A) as well as LPS from other bacteria of the Enterobacteriaceae family. Furthermore, penta-acylated LPS from Pseudomonas aeruginosa acted as an antagonist to hexa-acylated E. coli LPS, as did E. coli, as shown by its inhibitory effect on IL-8 production in stimulated cells. Hypo-acylated lipid A, such as that of P. aeruginosa, is found in several species within the gut microflora as well as in several bacteria causing chronic infections. Thus, our results suggest that the composition of the microflora may be important in modulating pro-inflammatory signalling in epithelial cells under normal as well as pathologic conditions. PMID- 14554247 TI - Enhanced cloning efficiency of mouse bone marrow macrophage progenitors correlates with increased content of CSF-1 receptor of their progeny at low oxygen tension. AB - Mononuclear phagocytes are located in every tissue of metazoan organisms. In this extravascular space, they are designated as macrophages and are known to sense and process many signals including the local oxygen tension (PO2), which ranges from 150 mmHg at the lung apices to around 40 mmHg in mixed venous blood and most organs, and to less than 10 mmHg in tissues where long-term and dynamic remodeling processes occur. Most tissue macrophages survive and maintain their differentiated status within an environment bathed by colony-stimulating factor (CSF)-1 through the CSF-1 receptor, encoded by the Csf1r gene. In order to investigate the mRNA expression profile of macrophages as a function of PO2, we developed an in vitro model in which monocyte-derived macrophages were generated from mouse bone marrow progenitor cells grown and maintained under low (36 mmHg) or atmospheric (142 mmHg) PO2, in the presence of L929-conditioned medium (L-CM) as a source of CSF-1. We show that CSF-1-reactive C57BL/6 bone marrow cells displayed an increased cloning efficiency under a PO2 of 36, compared with 142 mmHg. Furthermore, we provide evidence of the overexpression of both CSF-1 receptor protein and mRNA by mouse monocyte-derived macrophages generated from bone marrow under low PO2. PMID- 14554248 TI - Control of Mycobacterium bovis BCG infection with increased inflammation in TLR4 deficient mice. AB - Live mycobacteria have been reported to signal through several pattern recognition receptors (PRR), among them toll-like receptor 4 (TLR4) and TLR2 in vitro. Here, we investigated the role of TLR4 in host resistance to Mycobacterium bovis (BCG) infection in vivo. In vitro, macrophages of TLR4 mutant C3H/HeJ mice infected with BCG expressed lower levels of TNF than controls, and TNF release was further decreased, although not completely absent, in the absence of TLR2. In vivo, TLR4 mutant C3H/HeJ and control C3H/HeOUJ mice were infected with BCG (2 x 10(6) CFU i.v.). Both TLR4 mutant and wild-type mice were able to control the infection and survived 8 months post-BCG infection. Macrophage activation with abundant acid-fast bacilli and expression of inducible nitric oxide synthase (iNOS) and MHC class II antigens was seen in both groups of mice. However, TLR4 mutant mice experienced an arrest of body weight gain and showed signs of increased inflammation, with persistent splenomegaly, increase in granuloma number and augmented neutrophil infiltration. Infection of TLR4-deficient mice with higher doses of BCG (1 and 3 x 10(7) CFU, i.v.) increased the inflammation in spleen and liver, associated with a transient, higher bacterial load in the liver. In summary, TLR4 mutant mice show normal macrophage recruitment and activation, granuloma formation and control of the BCG infection, but this is associated with persistent inflammation. Therefore, TLR4 signaling is not essential for early control of BCG infection, but it may have a critical function in fine tuning of inflammation during chronic mycobacterial infection. PMID- 14554249 TI - The RD1 proteins of Mycobacterium tuberculosis: expression in Mycobacterium smegmatis and biochemical characterization. AB - A 9.5-kb section of DNA called region of deletion 1 (RD1) is present in virulent Mycobacterium tuberculosis strains but is deleted in all attenuated Mycobacterium bovis BCG vaccine strains. This region codes for at least nine genes. Some or all RD1 gene products may be involved in virulence and pathogenesis, and at least two, ESAT-6 and CFP-10, represent potent T- and B-cell antigens. In order to produce the entire set of RD1 proteins with their natural posttranslational modifications, a robust expression system for M. tuberculosis proteins in the fast-growing saprophytic strain Mycobacterium smegmatis was developed. Our system employs the inducible acetamidase promoter and allows translational fusion of recombinant M. tuberculosis proteins with polyhistidine or influenza hemagglutinin epitope tags for affinity purification. Using eGFP as reporter gene, we showed that the acetamidase promoter is tightly regulated in M. smegmatis and that this promoter is much stronger than the widely used constitutive groEL2 promoter. We then cloned 11 open reading frames (ORFs) found within RD1 and successfully expressed and purified the respective proteins. Sera from tuberculosis patients and M. tuberculosis-infected mice reacted with 10 purified RD1 proteins, thus demonstrating that Rv3871, Rv3872, Rv3873, CFP-10, ESAT-6, Rv3876, Rv3878, Rv3879c and ORF-14 are expressed in vivo. Finally, glycosylation of the RD1 proteins was analyzed. We present preliminary evidence that the PPE protein Rv3873 is glycosylated at its C terminus, thus highlighting the ability of M. smegmatis to produce M. tuberculosis proteins bearing posttranslational modifications. PMID- 14554250 TI - Diagnostic evaluation of 2', 5'-oligoadenylate synthetase activities and antibodies against Epstein-Barr virus and Coxiella burnetii in patients with chronic fatigue syndrome in Japan. AB - To investigate the association of viral infections with chronic fatigue syndrome (CFS), we assayed 2', 5'-oligoadenylate synthetase (2-5AS) activities in peripheral blood mononuclear cells from CFS patients in Japan. These patients were diagnosed in two hospitals, H1 and H2, located in different areas of the country. The activities were detected in 19 (86%) and 7 (32%) of each of the 22 patients in H1 and H2, respectively, while they were detected in only four (11%) out of the 38 healthy controls. IFN-alpha was similarly detected in a few CFS patients and healthy controls. We also assayed the antibody titers against Epstein-Barr virus (EBV) and Coxiella burnetii in these patients. The EBV anti-EA IgG antibodies were detected in two (9%) and seven (32%) of each of the 22 patients in H1 and H2, respectively. Anti-C. burnetii IgG antibodies were detected in six (27%) out of 22 patients in H1 but not in 22 patients in H2, while they were detected in one (11%) of the nine healthy controls. Some CFS patients may be associated with EBV or C. burnetii infection. There were some statistical correlations between the 2-5AS activities and antibody titers of EA IgG (P < 0.05, Student's t-test) but not to the antibody titers of C. burnetii. The up-regulation of 2-5AS activities suggests immunological dysfunctions with some virus infections in the CFS patients. Our results indicate that 2-5AS activities are useful for a diagnostic marker of CFS and for exploring the complicated pathogenesis of CFS. PMID- 14554251 TI - Visceral leishmaniasis in eastern Sudan: parasite identification in humans and dogs; host-parasite relationships. AB - In 1996, an epidemic outbreak of visceral leishmaniasis (VL) started in Barbar el Fugara, a village in Gedarif State (eastern Sudan). From 1997 to 2000, regular epidemiological studies were carried out in the human population, as well as in mammals and sand flies. In symptomatic patients, 46/69 lymph node, 6/20 post kala azar dermal leishmaniasis (PKDL) and 1/4 cutaneous cultures in NNN medium were positive. In 69 dogs, 23/79 lymph node cultures were positive. In other mammals (47 rodents, five donkeys, one mongoose and one monkey) spleen and/or blood cultures were negative. Characterization of isolated strains (by starch gel electrophoresis and isoelectrofocusing) identified three zymodemes of Leishmania donovani, two of L. infantum and two of L. archibaldi complexes from patient samples and three zymodemes of L. donovani, three of L. infantum and two of L. archibaldi complexes from dog samples. Five of them were present in both man and dog. For the first time, a strain from a PKDL case was identified as L. infantum, and a child had the same L. infantum zymodeme in VL and in subsequent PKDL. Blood samples from dogs were studied by immunofluorescent antibody test (IFAT). The seroprevalence in dogs was 72.5%, 74.3% and 42.9% in 1998, 1999 and 2000, respectively. By using CDC miniature light traps 12 745 sand flies were collected and then identified. Phlebotomus papatasi (7%) and P. orientalis (5%) were sympatric, mainly inside homes (85% and 75%, respectively). These results, the relative stability of seroprevalence in dogs and the intradomiciliar presence of P. orientalis, known as a vector of VL in Sudan, suggest several hypotheses: (i) man is responsible for the disease in dogs, (ii) the dog is the reservoir of VL, (iii) the dog is an intermediate host between a possible sylvatic cycle and the anthroponotic cycle. More extensive studies are needed to assess the transmission cycle of VL in this area of Sudan. PMID- 14554253 TI - The Syrian hamster as a model for the dilated cardiomyopathy of Chagas' disease: a quantitative echocardiographical and histopathological analysis. AB - Chronic Chagas' disease cardiomyopathy (CCC) is caused by the protozoan Trypanosoma cruzi, and it affects 30% of the 16-18 million people infected in Latin America. A good rodent model that develops a dilated cardiomyopathy closely resembling human CCC after T. cruzi infection is still needed. We compared the cardiomyopathy developed by T. cruzi-infected Syrian hamsters with human Chagas' disease cardiomyopathy using quantitative methods. Female hamsters were infected with 3.5 x 10(4) (G1, n = 10) or 10(5) (G2, n = 10) T. cruzi Y strain blood trypomastigotes. Control animals (C, n = 10) were injected with saline solution. Cardiac function was assessed by echocardiography at 4, 8 and 12 months post infection. Heart sections were submitted to histopathological/morphometric analysis 12 months post-infection. At this time, ventricular dysfunction and diffuse or multi-focal myocarditis were observed in 91% and 100% of G1 and G2 infected groups, respectively. Median interstitial collagen volumes in groups C, G1 and G2 were 1.2%, 1.9% and 3.9%, respectively, and were significantly higher in group G2 than in group C. Among infected animals, myocarditis showed a positive correlation with interstitial fibrosis. Deaths in the chronic phase (8 12 months post-infection) were more frequent among G2 than G1, and were associated with macroscopic ventricular dilation, severe myocarditis and increased fibrosis values, along with an earlier onset of ventricular dysfunction. The T. cruzi chronically infected Syrian hamster develops a cardiomyopathy which resembles human Chagas' disease cardiomyopathy, and might be an adequate tool to investigate pathogenic mechanisms of this disease and to search for novel therapeutic strategies. PMID- 14554252 TI - TH2 profile in asymptomatic Taenia solium human neurocysticercosis. AB - Neurocysticercosis (NC), a parasitic disease caused by Taenia solium, may be either asymptomatic or have mild to severe symptoms due to several factors. In this study, the immunological factors that underlie NC pleomorphism were studied. Ten of the 132 inhabitants of a rural community in Mexico (Tepez) had a computerized tomography (CT) scan compatible with calcified NC, and all were asymptomatic. Their immunological profiles were compared with those of 122 CT scan negative (non-NC) subjects from the same village. NC was associated with a TH2 response (IgG4, IL-4, IL-5, IL-13). Subjects from Tepez had higher levels of specific antibodies (IgG1, IgG2, IgG4, IgE) and specific cell proliferation than subjects from an area with low exposure (Ensenada). This suggests that non-NC subjects from Tepez had been exposed to T. solium and resisted infection in the brain. Distinct immunological profiles in equally exposed individuals differing in outcome of infection support the hypothesis of host-related factors in resistance to and pathogenesis of NC. This is the first study reporting the immunological profile associated with the asymptomatic form of NC. PMID- 14554255 TI - Usutu virus activity in Austria, 2001-2002. AB - Usutu virus (USUV), a member of the mosquito-borne clade within the Flaviviridae family, was responsible for avian mortality in Austria in 2001. In 2002, the virus continued to kill birds, predominantly blackbirds. High numbers of avian deaths were recorded within the city of Vienna and in surrounding districts of the federal state of Lower Austria, while single die-offs were noticed in the federal states of Styria and Burgenland. A total of 72 birds were submitted for laboratory examination, 30 of which tested positive for USUV by immunohistochemistry and/or polymerase chain reaction. Laboratory-confirmed cases of USUV infection originated from the federal states of Vienna and Lower Austria only. The data show that (i) USUV has managed to overwinter and has been able to establish a transmission cycle in Austria, (ii) the virus seems to have become a resident pathogen of Austria with a tendency to spread to other geographic areas, and (iii) the surveillance of dead blackbirds is a useful sentinel system for monitoring USUV activity. PMID- 14554254 TI - Characterization of experimental equine glanders. AB - Considerable advances in understanding of the disease caused by Burkholderia mallei have been made employing a combination of tools including genetic techniques and animal infection models. The development of small animal models has allowed us to assess the role of a number of putative virulence determinants in the pathogenesis of disease due to B. mallei. Due to the difficulties in performing active immunization studies in small animals, and due to the fact that the horse is the target mammalian species for glanders, we have initiated experimental studies on glanders in horses. Intratracheal deposition of B. mallei produced clinical glanders with organisms being recovered from tissues of infected horses. The model should prove to be of considerable value in our ongoing studies on the pathogenesis and vaccine development for glanders. PMID- 14554256 TI - Sorting out self and microbial lipid antigens for CD1. AB - CD1 proteins mediate T cell activation in response to self and foreign lipids, including lipid antigens from the intracellular pathogen Mycobacterium tuberculosis. During natural infections, myeloid cells migrate to sites of infection and use microbial pattern recognition receptors to internalize live bacteria and lipid antigens into the endosomal network. New studies show that certain CD1 proteins are particularly receptive to binding lipid antigens in the low pH environment of endosomes. Therefore, the endosomal network may represent a depot for concentrating and then selectively presenting exogenous foreign lipid antigens to T cells. PMID- 14554257 TI - Fascination with bacteria-triggered cell death: the significance of Fas-mediated apoptosis during bacterial infection in vivo. AB - Increasing evidence indicates that bacterial pathogens have developed mechanisms to modulate the apoptotic signaling cascade of host cells and thereby cause disease. The Fas death receptor pathway is one of the most extensively investigated apoptotic signaling pathways. In this review we discuss the role of Fas signaling during the interplay between bacterial pathogens and the host in vivo. PMID- 14554258 TI - Availability and use of molecular hydrogen as an energy substrate for Helicobacter species. AB - Molecular hydrogen is produced in the large intestine of animals due to the fermentation reactions of sugar catabolism. The gastric pathogen Helicobacter pylori and the liver pathogen Helicobacter hepaticus have the capacity to use molecular hydrogen as a respiratory substrate. The amount of the gas within tissues colonized by these pathogens is ample, and use of H2 significantly increases the stomach colonization ability of H. pylori. PMID- 14555179 TI - Localization and photoaffinity labelling of the levetiracetam binding site in rat brain and certain cell lines. AB - Levetiracetam (2S-(2-oxo-1-pyrrolidinyl)butanamide, KEPPRA, a novel antiepileptic drug, has been shown to bind to a specific binding site located in the brain (Eur. J. Pharmacol. 286 (1995) 137). To identify the protein constituent of the levetiracetam binding site in situ, we synthesized the photoaffinity label [3H]ucb 30889 ((2S)-2-[4-(3-azidophenyl)-2-oxopyrrolidin-1-yl]butanamide), a levetiracetam analog with higher affinity for the levetiracetam binding site. This radioligand was used to map the levetiracetam binding site within the brain and to study its cellular and subcellular distribution. Autoradiography experiments using [3H]ucb 30889 in rat brain revealed a unique distribution profile that did not match that of classical receptors known to be involved in the generation of epileptic seizures. There was a high level of binding in the dentate gyrus, the superior colliculus, several thalamic nuclei, the molecular layer of the cerebellum and to a lesser extent in the cerebral cortex, the striatum and the hypothalamus. The levetiracetam binding site was restricted to neuronal cell types, undifferentiated PC12 cells and was highly enriched in synaptic vesicles. [3H]ucb 30889 was also used in photoaffinity labelling studies and shown to bind covalently to a membrane protein with a molecular weight of approximately 90 kDa. PMID- 14555178 TI - Binding characteristics of [3H]ucb 30889 to levetiracetam binding sites in rat brain. AB - Levetiracetam (2S-(2-oxo-1-pyrrolidinyl)butanamide, KEPPRA, a novel antiepileptic drug, has been shown to bind to a specific binding site located in brain (levetiracetam binding site [Eur. J. Pharmacol. 286 (1995) 137]). However, [3H]levetiracetam displayed only micromolar affinity for these sites making it an unsuitable probe for further characterization. The present study describes the binding properties of an analogue of levetiracetam: [3H]ucb 30889, (2S)-2-[4-(3 azidophenyl)-2-oxopyrrolidin-1-yl]butanamide. [3H]ucb 30889 binds reversibly to specific binding sites in rat brain. Kinetics at 4 degrees C were biphasic with half-times of association and dissociation of, respectively, 3 and 4 min for the fast component and 47 and 61 min for the slow component. [3H]ucb 30889 saturation binding curves were compatible with the labelling of a homogenous population of binding sites having a B(max) of 4496+/-790 fmol/mg protein (mean+/-S.D., n=5) and a K(d) of 62+/-20 nM (mean+/-S.D., n=5), a 20-fold increase in affinity compared to [3H]levetiracetam. Competition binding curves with ligands known to interact with levetiracetam binding sites and tissue distribution restricted to the brain indicated that [3H]ucb 30889 and [3H]levetiracetam bind to the same site. Although levetiracetam binding sites and GABA(A) (gamma-aminobutyric acid) receptors share some ligands such as pentobarbital and pentylenetetrazol, experiments performed with [35S]TBPS (tert-butyl-bicyclo[2.2.2]phosporothionate), a probe for the GABA(A) Cl(-) channel do not support the hypothesis that levetiracetam binding sites are part of the GABA(A) receptor complex. Preliminary autoradiography studies in rat brain revealed that [3H]ucb 30889 labels specific sites in all brain regions and that this binding is concentration-dependently displaced by levetiracetam. PMID- 14555180 TI - Evidence for inhibitory effect of the agonist gaboxadol at human alpha 1 beta 2 gamma 2S GABAA receptors. AB - Gaboxadol (THIP; 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol) is an agonist at GABA(A) receptors. THIP concentrations (0.01-50 mM) were applied rapidly to Sf9 cells expressing the human alpha(1)beta(2)gamma(2S) GABA(A) receptors. The EC(50) values for the peak current in THIP alone or THIP plus 1 microM diazepam were 154 and 53 microM, respectively. In supersaturating THIP (10-50 mM) the rate of current decay increased and an off-current developed when THIP was rapidly removed. The mean currents measured over the first 4 s in 10 mM and higher THIP concentrations were 0.6 or less of the 1 mM THIP mean current. Diazepam (1 microM) increased the 4 s mean current when evoked by 10 to 20 mM THIP but not 50 mM THIP. No similar effects on the current time-course were recorded in supersaturating gamma-aminobutyric acid (GABA) concentrations (50 and 80 mM). The results demonstrate an inhibitory as well as agonist effect of THIP at alpha(1)beta(2)gamma(2S) GABA(A) receptors. PMID- 14555181 TI - High-throughput real-time monitoring of Gs-coupled receptor activation in intact cells using cyclic nucleotide-gated channels. AB - Cyclic adenosine-monophosphate (cAMP) is one of the major second messenger molecules transmitting extracellular stimuli into short- and long-term changes of intracellular homeostasis. Measurements of cellular cAMP levels are often used to quantify and characterize signaling by G protein-coupled receptors. Current assays for cAMP determination are usually end-point assays involving cell lysis. We have developed a technology to monitor real-time changes of cAMP levels in living cells. This method uses a modified cyclic nucleotide-gated (CNG) Ca(2+) channel which is opened by intracellular cAMP. Thus, changes in cAMP levels are translated into changes in free Ca(2+) which can easily be measured using fluorimetric imaging technologies compatible with high-throughput screening formats. The new assay method was used to characterize the pharmacology of various endogenously and heterologously expressed G protein-coupled receptors and allows for the simultaneous study of G(s), G(i) and G(q)-linked receptors in the same cell population. PMID- 14555182 TI - 17 Beta-estradiol increases endothelial nitric oxide synthase mRNA copy number in cerebral blood vessels: quantification by real-time polymerase chain reaction. AB - The enzyme endothelial nitric oxide synthase (eNOS) plays a critical role in the maintenance of vascular tone. The mechanism by which estrogen increases eNOS function remains controversial. We demonstrate here using real-time polymerase chain reaction (PCR) and immunoblot analysis that in vivo estrogen treatment leads to a 100% increase in eNOS messenger RNA (mRNA) copy number and increases eNOS protein levels by 47% in mouse cerebral blood vessels. These data suggest that estrogen can modulate eNOS at the transcriptional level in blood vessels in vivo. PMID- 14555183 TI - Phosphatidylinositol 3-kinase in angiotensin II-induced hypertrophy of vascular smooth muscle cells. AB - Activation of 4E-binding protein 1 (4E-BP1) by growth factors regulates protein synthesis in vascular smooth muscle cells. The interaction between G protein coupled receptors and activated 4E-BP1 is unclear. We examined phosphadityl inositol (PI) 3-kinase in angiotensin II-induced 4E-BP1 phosphorylation in cultured rat vascular smooth muscle cells. Angiotensin II time and dose dependently stimulated phosphorylation of 4E-BP1 through the angiotensin AT(1) receptor. Pretreatment with wortmannin or 2-(4-Morpholinyl)-8-phenyl-4H-1 benzopyran-4-one (LY294002), a PI 3-kinase inhibitor, suppressed angiotensin II induced phosphorylation, but a mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinases (ERK) kinase-1 (MEK-1) inhibitor, 2'-Amino-3'-methoxyflavone (PD98059), and a p38 MAPK inhibitor, 4-(4 Fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)1H-imidazole (SB203580), had no effect. With regard to the involvement of mammalian target of rapamycin (mTOR) and p70 S6 kinase, angiotensin II-induced phosphorylation was abolished by pretreatment with rapamycin, but not by tosylphenylalanine chloromethyl ketone or tosyllysine chloromethyl ketone. Ca(2+) was involved, since intracellular Ca(2+) chelation inhibited angiotensin II-induced phosphorylation while a Ca(2+) ionophore, A23187, stimulated phosphorylation. Thus, angiotensin II induces the phosphorylation of 4E-BP1 via the PI 3-kinase/mTOR pathway, but not via ERK or p70 S6 kinase. PMID- 14555184 TI - The kynurenine 3-hydroxylase inhibitor Ro 61-8048 improves dystonia in a genetic model of paroxysmal dyskinesia. AB - The effects of the novel kynurenine 3-hydroxylase inhibitor 3,4-dimethoxy-N-[4-(3 nitrophenyl)thiazol-2-yl]benzenesulfonamide (Ro 61-8048) on severity of dystonia were examined in dt(sz) mutant hamsters, an animal model of paroxysmal dystonia, in which stress precipitates dystonic episodes. Ro 61-8048 (50, 100 and 150 mg/kg i.p.) significantly reduced the severity of dystonia in dt(sz) hamsters without leading to marked central side effects. Determinations of kynurenic acid concentrations in brain homogenates demonstrated that Ro 61-8048 (100 mg/kg i.p.) provoked a two- to threefold increase of the endogeneous broad spectrum glutamate receptor antagonist kynurenic acid in the striatum, cerebellum and brainstem of mutant hamsters. The antidystonic efficacy of Ro 61-8048 at well-tolerated doses suggests that kynurenine 3-hydroxylase inhibitors should be considered as new therapeutic candidates for the treatment of dyskinesias. PMID- 14555185 TI - Thyroxine pretreatment increases basal myocardial heat-shock protein 27 expression and accelerates translocation and phosphorylation of this protein upon ischaemia. AB - Thyroxine pretreatment increases the tolerance of the heart to ischaemia, and heat-shock protein 27 (HSP27) is considered to play an important role in cardioprotection. The present study investigated whether long-term thyroxine administration can induce changes in the expression, translocation and phosphorylation of HSP27 at baseline and upon ischaemic stress. L-Thyroxine (T(4)) was administered to Wistar rats (25 microg/100 g/day s.c.) for 2 weeks, while normal animals served as controls. Hearts from normal and thyroxine-treated rats were perfused in Langendorff mode and subjected to 10 or 20 min of zero-flow global ischaemia only or to 20 min of ischaemia followed by 45 min of reperfusion. Total and phospho-HSP27 expression were assessed at different times in the Triton-soluble (cytosol-membrane), S fraction, and the Triton-insoluble (cytoskeleton-nucleus) fraction, P fraction. Postischaemic recovery of left ventricular developed pressure at 45 min of reperfusion was expressed as % of the initial value. In hearts from thyroxine-treated animals, the levels of basal total HSP27 and phospho-HSP27 in the P fraction were significantly increased as compared to normal. In response to ischaemia, in hearts from thyroxine-treated rats, the levels of total HSP27 and phospho-HSP27 were found to be significantly increased in the P fraction at 10 and 20 min of ischaemia as compared to preischaemic values, whereas in normal hearts, the levels of total HSP27 and phospho-HSP27 were significantly increased at 20 min only. Postischaemic functional recovery was significantly greater in thyroxine-treated than in untreated hearts. In summary, long-term thyroxine pretreatment results in an increased basal expression and phosphorylation of HSP27 and in an earlier and sustained redistribution of HSP27 from the S to the P fraction in response to ischaemia. This effect might be of important therapeutic relevance. PMID- 14555186 TI - Pharmacological characterization of YM598, an orally active and highly potent selective endothelin ET(A) receptor antagonist. AB - We describe here the pharmacology of (E)-N-[6-methoxy-5-(2-methoxyphenoxy)[2,2' bipyrimidin]-4-yl]-2-phenylethenesulfonamide monopotassium salt (YM598), a novel selective endothelin ET(A) receptor antagonist synthesized through the modification of the ET(A)/ET(B) non-selective antagonist, bosentan. YM598 inhibited [125I]endothelin-1 binding to cloned human endothelin ET(A) and ET(B) receptor, with K(i) of 0.697 and 569 nM, and inhibited endothelin-1-induced increases in intracellular Ca(2+) concentration in human and rat endothelin ET(A) receptor. YM598 also inhibited endothelin-1-induced vasoconstriction in isolated rat aorta with a pA(2) value of 7.6. In vivo, YM598 inhibited the pressor response to big endothelin-1, a precursor peptide of endothelin-1. DR(2) values of YM598 in pithed rats were 0.53 mg/kg, i.v. and 0.77 mg/kg, p.o., and its antagonism in conscious rats was maintained for more than 6.5 h at 1 mg/kg, p.o. In contrast, YM598 had no effect on the sarafotoxin S6c-induced depressor or pressor responses. YM598 showed not only superior antagonistic activity and higher-selectivity for endothelin ET(A) receptor in vitro, but at least a 30-fold higher potency in vivo than bosentan. In conclusion, YM598 is a potent and orally active selective endothelin ET(A) receptor antagonist. PMID- 14555187 TI - Effect of experimental diabetes on cholinergic, purinergic and peptidergic motor responses of the isolated rat bladder to electrical field stimulation or capsaicin. AB - An attempt has been made to pharmacologically isolate cholinergic, P(2) purinoceptor-mediated and peptidergic (capsaicin-sensitive, tachykinin-mediated) contraction of the guanethidine-treated rat bladder detrusor preparation, in vitro. The effect of experimental diabetes was assessed on these types of contraction. Responses were evoked by electrical field stimulation (single shocks or 1 Hz for 30 s or 10 Hz for 40 s). Single shocks and 1-Hz stimulation were applied in the presence of (a). atropine (1 microM) or (b). P(2) purinoceptor antagonists (50 microM pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid) [PPADS] plus 100 microM suramin. Long-term electrical field stimulation (10 Hz for 40 s) (c). was applied with both atropine and the P(2) purinoceptor antagonists present in the organ bath. The effects of capsaicin (d). and ATP (e). were also studied. Three groups of experimental animals were used: streptozotocin treated (50 mg.kg(-1) i.p., 8 weeks before the experiment), parallel solvent treated and untreated rats. (a). Responses to electrical field stimulation in the presence of atropine were reduced by half by PPADS plus suramin, but were resistant to capsaicin tachyphylaxis. They were enhanced in preparations taken from diabetic rats. (b). Contractions to electrical field stimulation in the presence of PPADS plus suramin were reduced by 2/3 by atropine, but were left unchanged by capsaicin or diabetes. (c). Contractions to long-term stimulation had a quick and a sustained phase. Especially the latter was inhibited by capsaicin tachypyhlaxis; it was also strongly reduced in preparations taken from diabetic rats. (d). Contractions to capsaicin (30 nM and 1 microM) were resistant to tetrodotoxin, strongly reduced by a combination of tachykinin NK(1) and NK(2) receptor antagonists, and slightly reduced in preparations from diabetic animals. Capsaicin (1 microM) had no acute inhibitory action on cholinergic or purinergic responses, nor did it cause relaxation in precontracted preparations treated with tachykinin receptor antagonists. (e) ATP-induced contractions were strongly reduced by PPADS plus suramin (50 plus 100 microM) and to a similar degree by 100 plus 200 microM, respectively. It is concluded that experimental diabetes selectively impairs peptidergic, capsaicin-sensitive responses (especially those that involve impulse conduction) in the rat detrusor preparation. The contractile response to electrical field stimulation that remains after atropine plus the P(2) purinoceptor antagonists has a yet unknown transmitter background. PMID- 14555188 TI - Disruption of oxygen-regulated responses underlies pathological changes in the placentas of women who smoke or who are passively exposed to smoke during pregnancy. AB - Previously, we showed that maternal smoking harms human placental development by changing the balance between cytotrophoblast (CTB) proliferation and differentiation. To understand the mechanisms involved, we studied the effects of maternal smoking and in vitro exposure of CTBs to nicotine and on CTB expression of molecules that govern cellular responses to oxygen tension: the von Hippel Lindau tumor suppressor protein (pVHL), the hypoxia-inducible transcription factors (HIFs), and the vascular endothelial growth factors (VEGFs). We previously reported that hypoxia upregulates CTB pVHL expression (1). Here we show that in vitro exposure of CTBs to nicotine has the same effect. Maternal smoking also dysregulated CTB expression of all three molecules. Remarkably, we found that passive exposure to cigarette smoke had many of the same effects as active smoking, a graphic demonstration of the ill effects of cigarette smoke, even secondhand, on placental development. Together, these findings explain, in part, how smoking damages the placenta by altering expression of key mediators of placental development. PMID- 14555189 TI - Lack of evidence for intergenerational reproductive effects due to prenatal and postnatal undernutrition in the female CD-1 mouse. AB - The impacts of adverse environments during the prenatal and/or early postnatal periods may be manifested as functional deficits that occur later in life. Epidemiological studies have shown an association of sub-optimal pregnancy outcomes in one generation with similar events in the following one, a phenomenon termed the "intergenerational effect". Data indicate that the incidence of adverse pregnancy outcomes and/or low birth weight infants is more closely correlated with the mother's perinatal environment than with that during her pregnancy. However, epidemiological studies are inherently limited given the variability of lifestyles, ethnicity, nutritional status, and exposures to environmental factors. An appropriate animal model would permit control of parameters that may be impossible to evaluate in human populations. The current studies investigated the mouse as a possible animal model. Pregnant CD-1 mice were placed on an ad libitum or food-restricted diet (50% normal) throughout gestation to generate control (CON) and intrauterine growth retarded (IUGR) litters. At birth (postnatal day (PD) 1) pups (F1) were cross-fostered to control dams in litters of either 8 (CON) or 16 (postnatal food restriction (FR)). The experimental groups thus generated represented adequate nutrition (CON-CON) and undernutrition during the prenatal (IUGR-CON), or postnatal periods (CON-FR), or both (IUGR-FR). Pups of dams on a restricted diet during gestation had significant IUGR (P<0.001) as compared to controls (birth weights of 1.32 g versus 1.63 g). At weaning, the average weight of the pups was dependent on postnatal litter size and the difference in birth weights between IUGR and CON animals was not a significant factor. CON-CON pup weight was 24.1g and IUGR-CON was 22.2 g as compared to the CON-FR (17.0 g) and IUGR-FR (17.3 g) groups. The difference in weaning pup weights between the FR and CON groups was significant (P<0.01). The F1 FR females did not reach CON female weights at any time point through 11 months after weaning. At PD60, a single breeding period for all groups of females with CON males began and continued for 75 days with 17 opportunities for breeding. Animals that became pregnant during this time were removed and allowed to litter. No significant differences were noted in average F2 litter size or average pup weight at birth: (CON-CON 12.2/1.62 g; IUGR-CON 11.9/1.6 2 g; CON-FR 10.9/1.70 g; IUGR-FR 11.3/1.61 g). We conclude that body weight at birth in the CD-1 mouse is not correlated with growth through the period of weaning (PD28). We did not find any evidence for an intergenerational reproductive effect after developmental undernutrition. PMID- 14555190 TI - Disruption of testicular steroidogenesis and epididymal function by inhaled benzo(a)pyrene. AB - The objective of this study was to evaluate the effect of sub-acute exposure to inhaled benzo(a)pyrene (BaP) on testicular steroidogenesis and epididymal function in Fisher 344 rats. Animals were assigned randomly to two control groups and one experimental group for each exposure regimen. Treatment consisted of sub acute exposure of rats via inhalation to 25, 75, and 100 microg BaP/m(3), 4 h daily for 10 days. Control animals were either exposed to carbon black (CB; sham) to control for inert BaP carrier or they remained unexposed (UNC). Blood samples were collected immediately after the cessation of exposures (time 0) and at 24, 48, and 72 h post-cessation of exposure, to assess the effect of bioavailable BaP on systemic testosterone and luteinizing hormone (LH) concentrations by radioimmunoassay (RIA). Progressive sperm motility of stored sperm (cauda epididymal sperm) was determined microscopically, while density of stored sperm was determined by hemocytometric counting. Progressive motility of stored sperm was reduced in rats exposed to 75 and 100 microg BaP/m(3) compared with their counterparts that were exposed to 25 microg BaP/m(3) or controls. Plasma testosterone concentrations declined as a result of exposure of rats to 75 microg BaP/m(3) from 0 to 48 h post-termination of exposure compared with controls (P<0.05; treatment x time interaction). This decrease was followed subsequently by a compensatory increase in the plasma concentrations of this steroid at 72 h post-cessation of exposures compared with previous time periods and controls (P<0.05). Increases in the mean plasma LH concentrations were observed in rats exposed to 75 microg BaP/m(3) compared with controls, throughout the time periods studied (P<0.05; treatment x time interaction). These data suggest that sub-acute exposure to inhaled BaP contributes to reduced testosterone concentrations and consequently impaired epididymal function of exposed animals. PMID- 14555191 TI - Elevated incidence of hypospadias in two sicilian towns where exposure to industrial and agricultural pollutants is high. AB - We found significant elevated incidence of hypospadias in two towns in Southeastern Sicily selected on the basis of the presence of intense industrial (Augusta) and agricultural (Vittoria) activities. Cases and controls were chosen in records collected from a surveillance system on abnormal live births in the same area and in a large city (Catania) located in an area at low risk of exposure to environmental pollutants. From 1991 to 1998, 16 cases of isolated hypospadias were recorded among male live births in Augusta (12.1 per 1000 male live births) and 24 cases in Vittoria (7.4 per 1000 male live births) with an incidence significantly higher than that expected (3.2 per 1000 in Southeastern Sicily). Relative risks in Augusta and Vittoria were 3.8 (95% confidence interval: 2.16-6.14) and 2.3 (95% confidence interval: 1.48-3.43; P=0.00003 and 0.04, respectively). In Augusta, the incidence of hypospadias was higher than in Vittoria. Significant log odds ratios were found for occupational exposure in fathers both in Augusta and Vittoria (P=0.0478 and 0.026, respectively). However, daily contact with pollutants in Augusta may not be sufficient by itself to determine hypospadias and other factors might be involved. Similar factors may act also in Vittoria. Thus, contact with large amounts of pesticides is, by itself, a risk factor for hypospadias, though genetic and other environmental factors might be involved. PMID- 14555192 TI - Assessment of male reproductive system in the CD-1 mice following oral manganese exposure. AB - Manganese has wide industrial applications and exposure to manganese can result in serious health conditions. The purpose of this study was to determine the reproductive effect of oral manganese exposure in male mice. Manganese acetate was tested at three dose levels (7.5, 15.0, and 30.0 mg/kg/day) for 43 days. The control group (0 mg/kg/day) received distilled water. Control negative group did not receive anything. Reproductive organ weights were recorded. Histopathology was performed on right testis, epididymis, seminal vesicle, and the accessory glands. Cauda epididymal, testicular sperm counts, and sperm motility was evaluated on the organ from the left side. The results of this study suggest that exposure to manganese caused a statistically significant (P<0.001) decrease in sperm motility and sperm counts at 15.0 and 30.0 mg/kg/day. There were no alterations in the fertility or pathology of the testicular tissue in the manganese-treated mice when compared with the controls. PMID- 14555193 TI - Cadmium-induced damage to primary cultures of rat Leydig cells. AB - The mechanism of testicular toxicity of cadmium is poorly understood. Previous studies focusing on cadmium-related changes in testicular histopathology have implicated testicular blood vessel damage as the main cause of cadmium toxicity. To further explore the toxic effects of cadmium on testis, we isolated and cultured rat Leydig cells, exposed to 10, 20, and 40 microM of cadmium chloride (base doses). After 24 h of exposure, cells and supernatants were harvested to examine cytotoxicity and genotoxicity of cadmium. The results show that both cell viability and concentration of testosterone excretion in primary Leydig cells are significantly lower in cadmium-exposed groups compared to the controls. Changes in testosterone excretion with human chorionic gonadotropin (hCG) stimulation is especially profound. The contents of malondialdehyde (MDA) and the activity of glutathione peroxidase (GSH-Px) in exposed groups are significantly higher than those in the control group, but the activity of superoxide dismutase (SOD) is lower. The number of cells with DNA single strand breaks and the levels of cellular DNA damage in all three exposure groups are significantly higher than in controls. These results indicate that cadmium is directly toxic to primary Leydig cells, and that the decreased percentage of normal cells and the increased level of DNA damage in cadmium-exposed Leydig cells may be responsible for decreased testosterone secretion. PMID- 14555194 TI - Ultrastructural modifications in the mitochondrion of mouse Sertoli cells after inhalation of lead, cadmium or lead-cadmium mixture. AB - CD-1 mice inhaled 0.01 M lead acetate, 0.006 M cadmium chloride or Pb-Cd mixture during 1h twice a week during 4 weeks. Testes were processed for transmission electron microscopic analysis. The percentage of damaged mitochondria was related to exposure time and the type of metal inhaled, noticing more damage when the mixture was administered. A dose-time relationship was found. Cadmium chloride caused the most severe mitochondrial alteration compared to lead acetate, whereas the mixture was more aggressive compared with each metal alone. Our results suggest that the changes in Sertoli cell could lead to a transformation process that may interfere with spermatogenesis. PMID- 14555195 TI - Modulation of ovarian follicle maturation in Long-Evans rats exposed to polychlorinated biphenyls (PCBs) in-utero and lactationally. AB - Polychlorinated biphenyls (PCBs) are ubiquitous man-made toxicants capable of endocrine disruption. Studies in several species have shown that exposure to PCBs and their hydroxylated metabolites reduces fecundity and decreases circulating concentrations of thyroid hormones, causing serious reproductive and developmental defects. Thyroid hormones modulate both follicular development and steroidogenesis, and affect estrogen metabolism and the regulation of estrogen receptor. This study was designed (1). to determine whether exposure to a commercially prepared PCB mixture (Aroclor 1016) exerts detrimental effects on follicle maturation in the Long-Evans hooded rat; and (2). to determine whether the modulatory effects of Aroclor can be attenuated by levo-thyroxine sodium (T(4)) supplementation. Animals were treated on gestation days 7-13 with a single daily intraperitoneal injection (2.5 mg/kg per day) of Aroclor. Half of the Aroclor-treated dams were also given T(4) supplements (2.89 microg/kg per day) via drinking water. Female pups were sacrificed on postnatal days 24/25, and the ovaries were excised, fixed for histology and analyzed. The analysis included a count, measurement and classification of healthy and atretic preantral and antral follicles in the greatest cross-sectional area. The results indicated that treatment with Aroclor significantly reduced the number of preantral follicles <50000 microm(2) and the total number of antral follicles in the 50-100000 and >100000 microm(2) size classes. T(4) circumvented the Aroclor effect on the number of preantral follicles <50000 microm(2); however, a significant reduction in the antral follicle number persisted in the 50-100000 and >100000 microm(2) size classes. In addition, we observed a significant increase in atresia in the Aroclor-treated ovaries in the antral <50000 microm(2) size class, which was not present in ovaries exposed to both Aroclor and T(4). These data support the hypothesis that Aroclor reduces the number of preantral and antral follicles of certain size classes in rats exposed during the critical period of development, and that supplementation with T(4) can attenuate the effects of Aroclor on small, but not medium or large antral follicles. Atresia of small, antral follicles may constitute one of the underlying mechanisms by which folliculogenesis is modulated by Aroclor 1016. PMID- 14555196 TI - Comparative embryotoxicities of butyl benzyl phthalate, mono-n-butyl phthalate and mono-benzyl phthalate in mice and rats: in vivo and in vitro observations. AB - The embryotoxic effects of butyl benzyl phthalate (BBP) and its two main metabolites mono-n-butyl (MBP) and mono-benzyl (MBzP) phthalate were evaluated in OF1 mice and Sprague-Dawley rats, in vivo and in whole embryo culture. In vivo, pregnant mice and rats received a single oral dose (0.9-5.4 mmol/kg) of either of these compounds on GD 8 and 10, respectively, and their fetuses were examined externally on GD 18 and 21, respectively. In mice, BBP, MBP and MBzP caused concentration-related embryolethality and malformations. In rats, MBP and MBzP did not show developmental toxicity. Some teratogenicity and a slight increase in post-implantation loss were observed after BBP administration, but mice were more susceptible to its toxic effects than were rats. In vitro, GD 8 mouse embryos and GD 10 rat embryos were cultured for 46 h in the presence of the test compounds (0.5 to 3-5mM). The cultured mouse embryos did not appear intrinsically more sensitive to MBP and MBzP, than the rat embryos. Altogether, these results suggest that the species sensitivity observed in vivo after an oral administration of BBP, MBP or MBzP during early organogenesis, might be due to maternal factors, i.e. toxicity and/or kinetics. PMID- 14555197 TI - Dibromoacetic acid-induced elevations in circulating estradiol: effects in both cycling and ovariectomized/steroid-primed female rats. AB - Oral exposures to high concentrations of the drinking water disinfection by product dibromoacetic acid (DBA) over the course of 14 days have been found to disrupt estrous cyclicity in the female rat. In order to investigate possible alterations in the relevant hormonal regulatory mechanisms, female Sprague-Dawley rats were gavaged for 2 weeks with 270 mg/kg DBA, ovariectomized (OVX) and implanted with estradiol capsules. For these females, the induced luteinizing hormone (LH) surge in these animals showed a borderline suppression in peak LH concentrations that was accompanied by a marked increase in circulating estradiol. This elevation in estradiol was DBA dose-related and, for intact, normally cycling females receiving lower doses of DBA (60 and 120 mg/kg, 14 days), was present on the day of estrus, at a time when a dramatic fall from proestrous concentrations is normally evident. Evaluations of liver microsomal cytochrome p450 activity in OVX/estradiol-implanted rats showed a suppression in ethoxyresorufin-O-deethylase (EROD) and pentoxyresorufin-O-deethylase (PROD) activity (indications of the activity of CYP1A and 2B, respectively-two key enzymes in estradiol oxidative metabolism). Phenobarbital (PhB) exposure in these animals did show induction of this activity, but was unable to lower E2 concentrations. This suggests that a DBA-induced suppression in estradiol catabolism is present and may either involve a targeted effect on the estrogen binding site on the CYP2B1/2 and CYP1A genes apart from the PhB-responsive unit, or a second pathway (possibly sulfation) that is not PhB-inducible. PMID- 14555198 TI - Effects of 4-tert-octylphenol, 4-tert-butylphenol, and diethylstilbestrol on prenatal testosterone surge in the rat. AB - In the present study, we evaluated the effects that 4-tert-octylphenol (OP) and 4 tert-butylphenol (BP) had on the prenatal testicular testosterone surge at embryonic day (ED) 19.5 in the rat. In utero exposure to alkylphenols (0.1-100 mg/kg maternal weight) on EDs 13.5, 15.5, and 17.5 did not decrease testicular testosterone content, whereas exposure to diethylstilbestrol (DES) caused a significant depression in testosterone synthesis and secretion. The depression was maintained during ex vivo tissue culture. In order to elucidate the observed differences in the in vivo effects between alkylphenols and DES, the exposures were also carried out in tissue culture of intact ED 19.5 testes. Basal testosterone, progesterone, cAMP production and hCG-induced testosterone levels were determined during and after a 3-h culture period. DES (100 mg/l) did not alter testosterone production but caused a two-fold increase in progesterone. OP (10, 100, 500 mg/l) and BP (100 mg/l) significantly increased testosterone and progesterone levels by up to seven-fold. In the presence of BP 100 mg/l, however, the intratesticular testosterone content did not correlate with the significantly increased fraction of secreted, or leaked, testosterone. The latter was correlated with tissue damage observed at electron microscopic level. Consistent with this, BP 500 mg/l elevated testicular testosterone level slightly during the first hour in the culture but the level subsequently returned to the control value. At the electron microscopic level, alkylphenols caused most severe changes in Leydig cell membrane structures and lipid droplets. In the DES-treated testes, membrane vesicle formation around the lipid droplets and increased mitochondrial pleiomorphy were observed. Altogether, the present in vivo and in vitro analyses confirm different effects of alkylphenols and DES on fetal rat steroidogenesis and tissue structure. PMID- 14555199 TI - Male reproductive effects of octylphenol and estradiol in Fischer and Wistar rats. AB - 4-tert-Octylphenol is a non-ionic surfactant used as a detergent, emulsifier and wetting agent. It is generally accepted that it acts as a weak estrogenic substance when evaluated in in vitro and in vivo short-term screening assays. The sensitivity of animal species (mouse versus rat), strain (inbred versus outbred) has been a matter of concern when selecting assay type for testing of estrogenicity of chemicals. The present study was designed to investigate whether the choice of different animal strain, could affect the outcomes of studies. Fischer and Wistar adult male rats were exposed to vehicle or 400 mg/kg bw of 4 tert-octylphenol administrated orally by gavage. Estradiol benzoate, at a dose of 40 microg/kg bw, was used as positive control agent. Treatment with estradiol benzoate decreased serum levels of testosterone, LH, FSH, inhibin and increased prolactin. Additionally, estradiol benzoate decreased the weight of all investigated reproductive organs, decreased sperm production and increased seminiferous tubular degeneration in both strains. More progressive effects on testis weight and histopathology were observed in the Fischer rats. Oral administration of octylphenol at 400 mg/kg bw to both rat strains increased prolactin levels but had no effect on LH, FSH, testosterone or inhibin. In the octylphenol-treated Fischer rats the weights of the seminal vesicles and the levator ani/bulbocavernosus muscle were significantly decreased, whereas only the levator ani/bulbocavernosus muscle was affected in Wistar rats. The weights of all other reproductive organs and sperm count were unaffected. It is concluded that there might be an organ specific difference in sensitivity between the two strains with the Fischer rat being the most sensitive rat model as demonstrated mainly by the more progressive effects on testis weight and histopathology in estradiol benzoate-treated Fischer rats but also by the decrease in seminal vesicle weight in octylphenol-treated rats. PMID- 14555200 TI - Histological and histomorphometric alterations in thyroid and adrenals of CD rat pups exposed in utero to methyl thiophanate. AB - Pregnant CD rats were treated with an initial dose of 0, 310 or 560 mg/kg bw per day of the fungicide methyl thiophanate (MT) on gestational days 10-14, corresponding to formation of thyroid and adrenal primordia; newborns were sacrificed on postnatal days (PNDs) 10 and 23. No apparent maternal toxicity and no effects on litter size, viability or weight gain were present. Delayed ear pinna detachment and eye opening were present at top dose level. Thyroid histology showed increased irregular nuclei and/or mitoses (PND 10-both doses), cells with necrotic or hydropic changes (PND 23-top dose). The adrenal cortex showed increased karyomegaly and hydropic degeneration (PND 23-both doses). Thyroid histomorphometry showed reduced follicular density, moderately increased follicular cell height and number of nuclei/follicle (PND 10-top dose and PND 23 both doses), suggesting retarded follicular maturation. The adrenal cortex relative area was slightly decreased (PND 10-top dose and PND 23-both doses).MT may act as weak endocrine disrupter, suggesting that attention should be paid to delayed endocrine alterations elicited by agrochemicals. PMID- 14555201 TI - Harmonization of rat fetal external and visceral terminology and classification. Report of the Fourth Workshop on the Terminology in Developmental Toxicology, Berlin, 18-20 April 2002. AB - This article is a report on the Fourth Berlin Workshop on Terminology in Developmental Toxicology, which was held in April 2002. The workshop is part of an international project in the field of harmonization of terminology in developmental toxicology supported by IPCS. The goal of the Harmonization Project is to ensure better chemical risk assessment. The aim of this Fourth Workshop was to discuss the results of a previously conducted survey on classification of external and visceral anomalies, which are listed in the international glossary, developed under the auspices of IFTS (1997 glossary). The discussions among experts from research institutions, regulatory agencies, and industries were mainly focussed on terms for which there was disagreement and/or uncertainties and the possible reasons. For the illustration of "gray-zone" anomalies, pictures were provided by the participants, which constituted the basis for detailed discussions. There was high agreement that most of the external anomalies (>66%) should be classified as malformations. The few external anomalies for which there was low agreement to classify as a malformation were discussed in detail. None of the external findings, which had in the survey a high agreement, were categorized as a variation.A high agreement regarding the classification of approximately one third of visceral anomalies was achieved with 34 and 2% being described as malformation and variation, respectively. Most of the visceral findings had low agreement indices and there appeared to be several reasons for this. Thus, the response, 'Not known/not used in the laboratory' (N) was often given. A couple of reasons for difficulties in the classification of an anomaly were that it is only rarely seen upon fetal examination or tends to be species specific. Furthermore, the classification of some anomalies as malformation or variation will remain vague as the decision must be made on a case-by-case basis. Factors affecting the decision include: the availability of appropriate historical control data, description of the grading and severity, whether the anomaly occurs in isolation or whether there is a relationship with an abnormal process, and finally, if the change represents an irreversible one, affecting human and/or animal health. It was concluded that a severity grading, supported by pictures of the anomaly, would be especially helpful to classify certain changes as malformation or as variation. Several of the soft tissue changes were considered likely to be the consequence of functional disorders and thus not strictly developmental anomalies. The possibility to describe a finding as 'Not Malformation' (Unclassified) was agreed upon. As a general conclusion it was emphasized that the observation of a permanent structural change should be considered to be a warning of possible consequences to humans, even when there is no apparent adverse effect on health and survival in adult animals of the species under investigation. Therefore, research is needed to further investigate postnatal consequences. Future collaboration in the field of reproductive and developmental toxicology should aim to further develop and implement a harmonized approach to the interpretation of study data. Therefore, this terminology work will continue in close cooperation with the IPCS Harmonization Project. A Steering Group should be established to facilitate the implementation of harmonized terminology into daily scientific work and its regulatory application. PMID- 14555204 TI - Mitochondrion-dependent caspase activation by the HIV-1 envelope. AB - Cells expressing the envelope glycoprotein complex (Env) encoded by the human immunodeficiency virus can fuse with cells expressing Env receptors (CD4 and CXCR4). The resulting syncytia undergo apoptosis. We developed a cytofluorometric assay for the quantitation of syncytium formation and syncytial apoptosis. Using this methodology, we show that caspase activation in syncytia is inhibited by pharmacological or genetic intervention on cyclin-dependent kinase-1, p53, and mitochondrial membrane permeabilization (MMP). Thus, transfection of fusing cells with the viral mitochondrial inhibitor of apoptosis encoded by cytomegalovirus, a specific inhibitor of MMP, prevented the mitochondrial cytochrome c release and abolished simultaneously the activation of caspase-3. Conversely, inhibition of caspases did not prevent MMP. These results indicate that Env-elicited syncytial apoptosis involves the intrinsic (mitochondrial) pathway. PMID- 14555205 TI - Mitochondrial state 3 to 4 respiration transition during Fas-mediated apoptosis controls cellular redox balance and rate of cell death. AB - The role of reactive oxygen species (ROS) production in death receptor-mediated apoptosis is ill defined. We show that ROS levels play a novel role in moderating the rate of cell death in Fas-dependent apoptosis. Treatment of Jurkat T cells with oligomycin (ATP-synthase inhibitor) or FCCP (mitochondrial uncoupler) and Fas activating antibody (CH11), facilitated rapid cell death. ATP levels, DEVDase activity and cytochrome c release were not account for the synergistic killing effect. However, a decrease in cellular ROS production was associated with CH11 treatment and combinations of CH11 with oligomycin or FCCP further inhibited cellular ROS levels. Thus, decreased ROS production is correlated with accelerated cell death. A transition from state 3 to state 4 mitochondrial respiration following apoptotic stimuli accounted for an attenuated membrane potential and as a results mitochondria-derived ROS production capacity diminished. Similar observations were demonstrated in isolated rat liver mitochondria. Transfection with mitochondrial targeted catalase inhibited mitochondrial ROS production and potentiated cell death. These data show that ROS production is important in receptor-mediated apoptosis and may play a pivotal role in cell survival. PMID- 14555206 TI - Regulation of endoplasmic reticulum Ca2+ dynamics by proapoptotic BCL-2 family members. AB - Uncontrolled cytosolic Ca(2+) overload is a common cause of cell death in several pathological conditions. Recent evidences reveal a more regulated role for intracellular Ca(2+) stores in controlling cell death. Proteins of the BCL-2 family include anti- and proapoptotic members that control the mitochondrial amplification loop of apoptosis. The antiapoptotic protein BCL-2 prevents this mitochondrial loop, while the "multidomain" proapoptotic proteins BAX and BAK are crucial to initiate it. BCL-2, BAX and BAK localize also to the endoplasmic reticulum (ER), the main intracellular Ca(2+) store. Overexpression of BCL-2 reduces resting ER Ca(2+) and death in response to apoptotic stimuli that mobilize Ca(2+). Our recent data indicate that multidomain proapoptotics also influence Ca(2+) metabolism. Cells deficient for Bax, Bak (DKO) display lowered steady state ER Ca(2+) concentrations ([Ca(2+)](er)) and secondarily decreased mitochondrial Ca(2+) uptake. Genetic and pharmacologic correction of [Ca(2+)](er) indicates that it controls death in response to Ca(2+)-dependent, mitochondria utilizing signals such as oxidative stress and lipid mediators; and that it participates in the regulation of the apoptotic response to most intrinsic stimuli, such as staurosporine. Thus, BAX and BAK control apoptosis not only at the mitochondria, but also at the ER, an obligate checkpoint for Ca(2+)-dependent apoptotic stimuli. PMID- 14555207 TI - Raf and the road to cell survival: a tale of bad spells, ring bearers and detours. AB - Research of the last years has demonstrated the absolute requirement of mitogenic signaling pathways for the control of cell survival. As reviewed here for the members of the Raf kinase family, apoptosis suppression proceeds through diverse mechanisms. They include the recruitment of novel effectors such as IAP and Bcl-2 proteins, key molecules in cell survival control, which interfere with the executions of the cell death at various levels, but also direct effects on metabolic events. PMID- 14555208 TI - Molecular mechanisms underlying WOX1 activation during apoptotic and stress responses. AB - Human WWOX gene encodes a putative tumor suppressor WW domain-containing oxidoreductase WOX1 (also known as WWOX or FOR). A high frequency of loss of heterozygosity (LOH) of this gene has been shown in prostate, lung, breast and other cancers. In addition, numerous aberrant WWOX mRNA transcripts have been found in cancer cells. WOX1 is a proapoptotic protein. In response to stress or apoptotic stimuli, WOX1 became phosphorylated at Tyr33, which enabled its complex formation with activated p53 and JNK1. The p53/WOX1 complex translocated to the mitochondria and further to the nuclei to mediate apoptosis. WOX1 mutants, which were inactivated for nuclear translocation or Tyr33 phosphorylation, failed to induce apoptosis, indicating that activation of WOX1 via Tyr33 phosphorylation, followed by nuclear translocation, is essential for inducing cell death. WOX1 induced apoptosis synergistically with p53. In contrast, transiently activated JNK1 induced anti-apoptotic response, and this protective activity inhibited WOX1 induced apoptosis. Taken together, WOX1 is involved in stress and apoptotic responses, and is likely to regulate the activation of both p53 and JNK1. PMID- 14555209 TI - Alternative pathways of ovarian apoptosis: death for life. AB - Ovarian cell death is an essential process for the homeostasis of ovarian function in human and other mammalian species. It ensures the selection of the dominant follicle and the demise of excess follicles. In turn, this process minimizes the possibility of multiple embryo development during pregnancy and assures the development of few, but healthy embryos. Degeneration of the old corpora lutea in each estrus/menstrual cycle by programmed cell death is essential for maintaining the normal cyclicity of ovarian steroidogenesis. Although there are multiple pathways that can determine cell death or survival, crosstalk among endocrine, paracrine and autocrine factors, as well as among protooncogenes, tumor suppressor genes, survival genes and death genes, play an important role in determining the fate of ovarian somatic and germ cells. The establishment of immortalized rat and human steroidogenic granulosa cell lines and the investigation of pure populations of primary granulosa cells allows for systematic studies of the mechanisms that control steroidogenesis and apoptosis of granulosa cells. We have discovered that during initial stages of granulosa cell apoptosis progesterone production does not decrease. In contrast, we found that it is elevated for up to 24hr following the onset of the apoptotic stimuli exerted by starvation, cAMP, p53 or tumor necrosis factor alpha stimulation, before total cell collapse. These observations raise the possibility for an alternative unique apoptotic pathway, one that does not involve mitochondrial cytochrome C release associated with the destruction of mitochondrial structure and steroidogenic function. Using mRNA from apoptotic cells and Affymetrix DNA microarray we discovered that Granzyme B, a protease that normally resides in T cytotoxic lymphocytes and natural killer cells of the immune system is expressed and activated in granulosa cells, thereby allowing the apoptotic signals to bypass mitochondrial signals for apoptosis, which can preserve their steroidogenic activity until complete cell destruction. This unique apoptotic pathway assures the cyclicity of estradiol and progesterone release in the estrus/menstrus cycle even during the initial stage of apoptosis. PMID- 14555210 TI - Basic fibroblast growth factor (bFGF) regulation of the plasma membrane calcium ATPase (PMCA) as part of an anti-apoptotic mechanism of action. AB - Basic fibroblast growth factor (bFGF) preserves the viability of at least 13 different cells, including epithelial, endothelial, smooth muscle and neuronal cells. In spite of this profound and rather universal effect on cell viability, detailed studies regarding the mechanism of bFGF's action have not been conducted. Rather, most studies have simply shown that bFGF inhibits cells from undergoing programmed cell death (i.e. apoptosis). The most mechanistic studies to date have been conducted on either neurons or ovarian (granulosa) cells. These studies have shown that bFGF prevents apoptosis through both genomic and acute actions. Basic FGF's acute actions involved the maintenance of normal levels of intracellular free calcium levels ([Ca(2+)](i)). In granulosa cells, bFGF maintained [Ca(2+)](i) through a protein kinase C(delta) (PKCdelta)-dependent mechanism. Further, bFGF-activated PKCdelta maintained [Ca(2+)](i) by stimulating calcium efflux. The ability of bFGF to stimulate calcium efflux involved the plasma membrane calcium ATPase (PMCA). Interestingly, bFGF-activated PKCdelta appeared to regulate PMCA activity in part by promoting its membrane localization. PMID- 14555211 TI - Potentiation of cellular antioxidant capacity by Bcl-2: implications for its antiapoptotic function. AB - A substantial body of data from clinical and laboratory studies indicates that reactive oxygen intermediates are implicated in the pathogenesis of diverse human diseases, including cancer, diabetes, and neurodegenerative disorders. Oxidative stress induced by reactive oxygen intermediates often causes cell death via apoptosis that is regulated by a plenty of functional genes and their protein products. Bcl-2, which is an integral intermitochondrial membrane protein, blocks apoptosis induced by a wide array of death signals. In spite of extensive research, the molecular milieu that characterizes the antiapoptotic function of Bcl-2 is complex and not fully identified. Recently, there are several lines of evidence that Bcl-2 functions via antioxidant pathways to prevent apoptosis. Thus, bcl-2-overexpressing cells exhibit elevated expression of antioxidant enzymes and higher levels of cellular GSH compared with the control cells transfected with the vector alone. There has been increasing evidence supporting that the redox-sensitive transcription factor nuclear factor kappaB regulates the activity and/or expression of antioxidative and antiapoptotic target genes and promotes cell survival against oxidative cell death. This commentary focuses on the antioxidative functions of Bcl-2 and underlying molecular mechanisms in relation to its antiapoptotic property. The role of Bcl-2 in regulation of nuclear factor kappaB signaling pathways and possible cross-talk with mitogen activated protein kinases are also discussed. PMID- 14555212 TI - Peroxisome proliferator-activated receptor gamma (PPARgamma) ligands as bifunctional regulators of cell proliferation. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma), a member of the ligand-activated nuclear receptor superfamily, plays a key role in mediating differentiation of adipocytes and regulating fat metabolism. PPARgamma has been implicated in the pathophysiology of atherosclerosis, inflammation, obesity, diabetes, immune response, and ageing. Recently, it has been shown that activation of PPARgamma by J(2) series cyclopentenone prostaglandins (cyPGs), especially 15-deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)) or synthetic agents, such as antidiabetic thiazolidinediones, causes anti-proliferation, apoptosis, differentiation, and anti-inflammation of certain types of cancer cells. The anti-proliferative effects of PPARgamma activators are associated with de novo synthesis of proteins involved in regulating the cell cycle and cell survival/death. Anti-inflammatory effects of 15d-PGJ(2) are associated with interruption of nuclear factor-kappaB and subsequent blockade of inflammatory gene expression. Furthermore, 15d-PGJ(2) at nontoxic doses induce expression of phase II detoxification or stress-responding enzymes, which may confer cellular resistance or adaptation to oxidative stress. The presence of a reactive alpha,beta-unsaturated carbonyl moiety in the cyclopentenone ring of 15d-PGJ(2) is important for part of biological functions this cyPG has. Recently, attention has been focused on the anti-proliferative activity of nonsteroidal anti inflammatory drugs (NSAIDs) in cancerous or transformed cells, which is mediated through interaction with PPARgamma irrespective of their ability to inhibit COX 2. Despite the fact that abnormally elevated COX-2 is associated with resistance to cell death, induction of apoptosis by certain NSAIDs is accompanied by up regulation of COX-2 expression. This commentary focuses on dual effects of the typical PPARgamma agonist 15d-PGJ(2) on cell proliferation and growth, and its possible involvement in the NSAID-induced COX-2 expression and apoptosis. PMID- 14555213 TI - Pleiotropic anti-apoptotic activity of glucocorticoids in ovarian follicular cells. AB - Glucocorticoids (GC) such as hydrocortisone and dexamethasone (DEX) protect steroidogenic granulosa cells against apoptosis induced by serum deprivation, cAMP, tumor necrosis factor alpha stimulation or p53 activation. The protective effects were evident both in primary rat and human granulosa cells, which comprise the main population of the ovarian follicular cells, as well as in steroidogenic granulosa cell lines established in our laboratory. A correlation between the expression of Bcl-2 protein and protection against apoptosis induced by DEX was found in granulosa cell lines expressing various levels of Bcl-2. Incubation with DEX leads to development of a rigid network of actin cytoskeleton and increased incidence of adherence and gap junctions. Higher content of connexin 43 and total cadherins were found in GC stimulated cells compared to non stimulated, suggesting that cell contact and intracellular communication contribute to the DEX induced resistance to apoptotic signals. Activation by DEX of MAPK and Akt/PKB but not p38 supported the view of a pleiotropic action of GC against apoptotic signals. Granzyme B, a protease characteristic for induction of apoptosis by T-cytotoxic lymphocytes and natural killer cells, was expressed and augmented during stimulation of apoptosis in the granulosa cells, and its synthesis and activation was blocked by DEX. It is concluded that GC exerted their anti-apoptotic effects in granulosa cells by multiple characteristic pathways. Moreover, the presence of endogenous granzyme B in granulosa cells suggest a novel intrinsic alternative apoptotic pathway that was earlier reported to be mediated uniquely by T-cytotoxic lymphocytes and natural killer cells. The anti-apoptotic effect of GC may play an important role in the healing process of the ovulatory follicle subsequent to follicular rupture and its rapid conversion to an active corpus luteum. PMID- 14555215 TI - Nuclear factor-kappa B plays a central role in tumour necrosis factor-mediated liver disease. AB - Deregulation of the apoptotic program is considered an important cause in liver disease. It became clear that the cytokine tumour necrosis factor (TNF) is of specific interest in this context. Therefore, from a clinical point of view, therapeutic control of TNF-receptor signalling pathways is highly desirable. These TNF-initiated signalling pathways result in a direct apoptotic response as well as potent activation of proinflammatory gene expression via activation of the transcription factor nuclear factor-kappa B (NF-kappaB). Since the latter pathway contributes to a series of liver pathologies, inhibition of hepatic NF kappaB activation was viewed as a potential therapy for liver injury. However, the more recent finding that NF-kappaB activation in hepatocytes is anti apoptotic shows that NF-kappaB signalling represents a problematic therapeutic target. Here we review the role of TNF and NF-kappaB in liver pathophysiology, and the underlying mechanisms of hepatocyte sensitisation to TNF toxicity in vivo. Based on this knowledge, we suggest some potential strategies for the treatment of TNF-mediated liver disease. PMID- 14555214 TI - Regulation of proliferation, survival and apoptosis by members of the TNF superfamily. AB - Tumor necrosis factor (TNF) was first identified in 1984 as a cytokine with anti tumor effects in vitro and in vivo. Extensive research since then has shown that there are at least 18 distinct members of the TNF super family and they exhibit 15-25% amino acid sequence homology with each other. These family members bind to distinct receptors, which are homologous in their extracellular domain. These cytokines have been implicated in a wide variety of diseases including tumorigenesis, septic shock, viral replication, bone resorption, rheumatoid arthritis, diabetes, and other inflammatory diseases. TNF blockers have been approved for human use in treating some of these conditions in the United States and other countries. Various members of the TNF super family mediate either proliferation, survival, or apoptosis of cells. Although distinct receptors, all members share a common cell signaling pathway that mediates the activation of nuclear factor-kappaB (NF-kappaB) and mitogen-activated protein kinases (e.g. c jun N-terminal kinase). Regulation of cell growth and activation of NF-kappaB and of c-jun N-terminal kinase by the TNF super family is mediated through sequential activation/association of a set of cell signaling proteins named TNF receptor associated factors, Fas-associated death domain and FADD-like ICE, caspases, receptor-interacting protein, NF-kappaB-inducing kinases, and IkappaBalpha kinases. Both apoptotic and antiapoptotic signals are activated simultaneously by the same cytokine in the same cell. Together these cytokines regulate cell growth/survival/apoptosis in a complex dance of changing partners and overlapping steps. PMID- 14555216 TI - Slowly getting a clue on CD95 ligand biology. AB - Since the ligand for the death factor CD95 (CD95L) was identified almost a decade ago, it has been established that this molecule (CD95L, FasL, Apo-1L, CD178, TNFSF6, APT1LG1) has multiple immunoregulatory and pathophysiologically relevant functions. CD95L does not only act as a death factor when externalized with secretory lysosomes on cytotoxic T and NK cells or when expressed on CD4(+) T cells in the course of activation-induced cell death, it is also a key molecule for the establishment of immune privilege or tumor cell survival and may serve as a costimulatory molecule during T cell activation. Moreover, alterations of expression or shedding of different forms of CD95L are associated with many diseases including various malignancies, HIV infection, autoimmune disorders (systemic lupus erythematodes, rheumatoid arthritis), acute myocardial infarction, traumatic injury and many others. In most cases, however, the physiological link between altered CD95L expression and pathophysiology is unknown. Given the potency of the molecule to regulate death and survival of many different cell types, the control of CD95L production, transport, storage, shedding and release is of tremendous biological and clinical interest. This commentary aims at briefly summarizing the current knowledge, hypotheses and controversies about CD95L as a multifunctional ligand and receptor. It touches upon the complex networks of intracellular dynamics of protein transport and trafficking and the potential bidirectional signal transduction capacity of CD95L with a focus on molecular interactions that have been worked out over the past years. PMID- 14555217 TI - TWE-PRIL; a fusion protein of TWEAK and APRIL. AB - TWEAK and APRIL are both members of the tumor necrosis factor family, which are involved in respectively angiogenesis and immune regulation. While TWEAK is processed at the cell surface, APRIL is processed inside the cell by a furin convertase and is solely able to perform its function as a soluble factor. Recently, TWE-PRIL has been identified, which is an endogenous hybrid transcript between TWEAK and APRIL. TWE-PRIL is a transmembrane protein that consists of a TWEAK intracellular, transmembrane and stalk region combined with APRIL as its receptor-binding domain. As such TWE-PRIL is expressed at the cell surface. Although TWE-PRIL, like APRIL, can stimulate T and B cell lines, distinct biological functions that may result from its membrane anchoring cannot be excluded. Understanding the function of this newly identified protein will contribute to the elucidation of the complexity of the tumor necrosis factor family. PMID- 14555218 TI - Apoptotic signal transduction pathways in diabetes. AB - Failure of insulin producing pancreatic beta-cells is a common characteristic of type 1 (insulin-dependent) and type 2 (insulin non-dependent) diabetes mellitus. Accumulating evidence suggests that programmed cell death (apoptosis) is the main form of beta-cell death in these disorders. The beta-cell is particularly sensitive to apoptotic stimuli due to the inherent features of the specialized beta-cell phenotype. In type 1 diabetes anti-beta-cell autoimmune reactivity delivers the apoptotic signals in the form of inflammatory mediators or T-cell effectors. In type 2 diabetes, the metabolic derangement is associated with production of inflammatory mediators in insulin-sensitive tissues leading elevated levels of circulating inflammatory mediators such as IL-6 and TNF. Further glucose has been suggested to induce beta-cell apoptosis via the induction of beta-cell synthesis of IL-1 which via autocrine action may elicit signalling cascades analogous to those seen in beta-cell destruction in type 1 diabetes. Considering the apparent importance of IL-1-beta signalling in beta cell failure in both type 1 and type 2 diabetes, we here review the modulatory effect exerted on IL-1signalling by cellular characteristics related to the specialized beta-cell phenotype. We conclude that beta-cell differentiation signals (Pdx-1), glucose metabolism, calcium handling as well as regulation of naturally occurring inhibitors of cytokine signalling contribute to sensitize the beta-cell to apoptotic stimuli. We hypothesize that immunological stimuli in type 1 diabetes and metabolic/inflammatory signals in type 2 diabetes converge on common signalling pathways leading to beta-cell failure and destruction in these two diseases. PMID- 14555219 TI - The putative role of apoptosis-modified histones for the induction of autoimmunity in Systemic Lupus Erythematosus. AB - In recent years, it has become evident that Systemic Lupus Erythematosus (SLE) is a disease characterized by an array of autoantibodies directed against the native nucleosome, its DNA component and/or its histone component. Nuclear antigens are generated and released in vivo during apoptosis. A hallmark of apoptosis is the cleavage of chromatin by caspase-activated DNase. This fragmentation occurs at the internucleosomal level and leads to DNA ladder formation classically associated with apoptosis. Thus, dysregulation of DNA fragmentation might be directly linked to the induction of autoimmunity in SLE. In our studies, activated human lymphoblasts contain high amounts of core histones in their cell lysates after apoptosis induction. This accumulation correlated highly with markers of early apoptosis (Annexin V positive, propidium iodide negative), but not with markers of late apoptosis or necrosis. Interestingly, accumulation of core histones or nucleosomes in cell lysates was detected as early as 30 or 60 min after UV irradiation, whereas phosphatidylserine externalization occurred 2 hr after apoptosis induction. Our results suggest that extranuclear accumulation of core histones is a very early event in apoptosis, preceding the externalization of phagocytosis signals on the outer membrane surface of apoptotically dying lymphoblasts. The following review will discuss these results in a broader perspective which includes our hypothesis of how apoptosis dysregulation during early phases may contribute to the induction of autoimmunity against nuclear autoantigens as seen in SLE. PMID- 14555221 TI - Non-apoptotic functions of caspases in cellular proliferation and differentiation. AB - The cysteinyl aspartate-specific proteases (caspases) have been identified as key players in the cellular process termed programmed cell death or apoptosis. During apoptosis, activated apoptotic caspases cleave selected target proteins to execute cell death. Additionally to their established function in cell death, a variety of recent publications have provided increasing evidence that apoptotic caspases also participate in several non-apoptotic cellular processes. Activated caspases exhibit functions during T-cell proliferation and cell cycle regulation, but are also involved in the differentiation of a diverse array of cell types. In some cell types, their differentiation can be morphologically viewed as a kind of incomplete apoptosis. Analysis of well-known apoptotic targets of caspases implicates that the cleavage of a limited number of selected substrates plays a major role during non-apoptotic functions of caspases. Selective substrate cleavage might be regulated by activation of anti-apoptotic factors, via a compartmentalized activation of caspases, or through limited activity of caspases during apoptosis-independent functions. The increasing evidence for caspase function in non-apoptotic cellular events suggests that caspases play a much more diverse role than previously assumed. PMID- 14555220 TI - The prodigiosins, proapoptotic drugs with anticancer properties. AB - The family of natural red pigments, called prodigiosins (PGs), characterised by a common pyrrolylpyrromethene skeleton, are produced by various bacteria. Some members have immunosuppressive properties and apoptotic effects in vitro and they have also displayed antitumour activity in vivo. Understanding the mechanism of action of PGs is essential for drug development and will require the identification and characterisation of their still unidentified cell target. Four possible mechanisms of action have been suggested for these molecules: (i) PGs as pH modulators; (ii) PGs as cell cycle inhibitors; (iii) PGs as DNA cleavage agents; (iv) PGs as mitogen-activated protein kinase regulators. Here, we review the pharmacological activity of PG and related compounds, including novel synthetic PG derivatives with lower toxicity and discuss the mechanisms of action and the molecular targets of those molecules. The results reported in this review suggest that PGs are a new class of anticancer drugs, which hold out considerable promise for the Pharmacological Industry. PMID- 14555222 TI - Apoptosis in the normal and inflamed airway epithelium: role of zinc in epithelial protection and procaspase-3 regulation. AB - The epithelium lining the airways is a physical barrier as well as a regulator of physiological and pathological events in the respiratory system. Damage to the epithelium by oxidants released from inflammatory cells is a critical factor in the pathogenesis of airway inflammatory diseases such as bronchial asthma. In these diseases, excessive apoptosis may be a likely mechanism responsible for damage to, and sloughing, of airway epithelial cells. Factors that increase the airway epithelium's resilience to apoptosis are likely to lessen the severity of this disease. One such factor is the dietary metal zinc. A special role for labile intracellular pools of zinc as anti-apoptotic agents in the regulation of the caspases, has emerged over the past two decades. This review focuses on caspase-inhibitory functions of zinc in airway epithelial cells, apparent abnormalities of zinc homeostasis in asthmatics and studies from the authors' laboratory which showed that zinc was strategically localized in the apical cytoplasm of airway epithelium to control caspase-3 activated apoptosis. These findings are discussed in the context of recent data from a murine model of allergic asthma, showing that loss of airway epithelial zinc was accompanied by changes in levels of both procaspase-3 and active caspase-3 and that nutritional zinc deprivation further increased airway epithelial apoptosis. We hypothesize that zinc has a protective role for the airway epithelium against oxyradicals and other noxious agents, with important implications for asthma and other inflammatory diseases where the epithelial barrier is vulnerable and compromised. PMID- 14555223 TI - In the cut and thrust of apoptosis, serine proteases come of age. AB - Proteolysis is central to the systematic cellular degradation that occurs during apoptosis. Predominantly, caspases have been studied in this regard. However, increasing evidence suggests that certain serine proteases may also play a significant role in apoptosis. Not only are these serine proteases involved in apoptosis signalling pathways independently, but they may also interact with more classical mediators of apoptosis such as the caspases or Bcl-2 family proteins. Isolation of apoptosis-associated serine proteases and the use of specific inhibitors have helped to shed light on potential pathways in which they are involved. Despite the recent developments in the field, knowledge regarding the role of serine proteases in apoptosis remains limited, but it is clear that investigations are gathering momentum and such studies may herald a new and exciting departure in apoptosis research. PMID- 14555224 TI - Induction of apoptosis by curcumin: mediation by glutathione S-transferase P1-1 inhibition. AB - Expression of glutathione S-transferase P1-1 (GSTP1-1) is correlated to carcinogenesis and resistance of cancer cells against chemotherapeutic agents. Curcumin, a natural compound extracted from Curcuma longa, has shown strong antioxidant and anticancer properties and also the ability to regulate a wide variety of genes that require activating protein 1 and nuclear factor kappaB (NF kappaB) activation. In the present study, we examined the inhibitory effect of curcumin on the expression of GSTP1-1 mRNA as well as protein, and we correlated this inhibition with the apoptotic effect of curcumin on K562 leukemia cells. Curcumin efficiently inhibited the tumour necrosis factor alpha- and phorbol ester-induced binding of AP-1 and NF-kappaB transcription factors to sites located on the GSTP1-1 gene promoter. TNFalpha-induced GSTP1-1 promoter activity was also inhibited by curcumin as shown by reporter gene assay. In parallel, curcumin induced pro-caspases 8 and 9 as well as poly ADP ribose polymerase cleavage and thus leading to apoptosis in K562 cells. Our results overall add a novel role for curcumin as this chemoprotective compound could contribute to induce apoptosis by its ability to inhibit the GSTP1-1 expression at the level of transcription. PMID- 14555225 TI - Cholesterol crystallization and macrophage apoptosis: implication for atherosclerotic plaque instability and rupture. AB - The presence of abundant cholesterol crystals symbolizes the disorder of cholesterol metabolism during the development of atherosclerosis. Examination of cultured human THP-1 macrophages treated with the cholesterol oxide, 7 ketocholesterol, revealed a concentration- and time-dependent increase in formation of cholesterol crystals in the cells. Radioisotope labeling and X-ray diffraction confirmed the presence of 7-ketocholesterol crystalline domains (d space 35.8A). Under the normal cell culture condition (5% CO(2), 37 degrees ), incubation with 7-ketocholesterol induced moderate levels of apoptosis. Elevating temperature from 37 to 40 degrees markedly reduces formation of the crystals in the macrophages. Meanwhile, at high temperatures, significantly increased numbers of apoptotic cells were detected in the cells treated with 7-ketocholesterol but not in those with native free cholesterol. These results suggest that hyperthermia inhibits cholesterol crystallization and promotes apoptotic effects of oxysterols on macrophages. PMID- 14555226 TI - Regulation of smooth muscle cell growth, function and death in vitro by activated mast cells--a potential mechanism for the weakening and rupture of atherosclerotic plaques. AB - The fibrous cap of a lipid-containing atherosclerotic plaque consists of collagen produced by arterial smooth muscle cells (SMCs) of synthetic phenotype. A thick cap protects the lipid-rich core, whereas a thin cap predisposes it to rupture, with ensuing acute clinical complications, such as myocardial infarction. Among the pathological mechanisms leading to plaque weakening and rupture, one possibility is loss of the matrix-synthesizing SMCs. Indeed, caps of ruptured coronary plaques contain a reduced number of SMCs. In contrast, in such lesions, the number of activated inflammatory cells, such as mast cells, is increased, suggesting that they may regulate the SMC number. We have shown that heparin proteoglycans secreted by activated mast cells can efficiently inhibit proliferation of SMCs in vitro and reduce their ability to produce collagen. Chymase, a neutral serine protease secreted by activated mast cells, can also inhibit SMC-mediated collagen synthesis by a transforming growth factor-beta dependent and -independent mechanism, and moreover, cause degradation of the collagen matrix by activating latent interstitial collagenase (MMP-1). Furthermore, chymase can induce SMC apoptosis by degrading the extracellular matrix component fibronectin necessary for SMC adhesion, with subsequent disruption of focal adhesions and loss of outside-in survival signaling. Thus, activated mast cells may participate in the weakening and rupture of atherosclerotic plaques by secreting mediators, such as heparin proteoglycans and chymase, which affect the growth, function and death of arterial SMCs. PMID- 14555227 TI - The changing faces of glutathione, a cellular protagonist. AB - Glutathione (GSH) has been described for a long time just as a defensive reagent against the action of toxic xenobiotics (drugs, pollutants, carcinogens). As a prototype antioxidant, it has been involved in cell protection from the noxious effect of excess oxidant stress, both directly and as a cofactor of glutathione peroxidases. In addition, it has long been known that GSH is capable of forming disulfide bonds with cysteine residues of proteins, and the relevance of this mechanism ("S-glutathionylation") in regulation of protein function is currently receiving confirmation in a series of research lines. Rather paradoxically, however, recent studies have also highlighted the ability of GSH-and notably of its catabolites-to promote oxidative processes, by participating in metal ion mediated reactions eventually leading to formation of reactive oxygen species and free radicals. A crucial role in these phenomena is played by membrane bound gamma-glutamyltransferase activity. The significance of GSH as a major factor in regulation of cell life, proliferation, and death, should be regarded as the integrated result of all these roles it can play. PMID- 14555228 TI - Anti-apoptotic effect of HIV protease inhibitors via direct inhibition of calpain. AB - Treatment with drugs designed to inhibit the HIV protease ameliorates immune functions in AIDS patients, reducing cell deletion by apoptosis even in the absence of inhibition of viral spread. This suggests that they interact with the intrinsic apoptotic signaling. We found that caspases, the main executioner of the apoptotic process, are not directly inhibited. In search for the mechanism responsible for their anti-apoptotic effect, we have found that indinavir and ritonavir are able to inhibit apoptosis only in those cell systems where apoptosis involves the activation of calpains. They directly inhibit a calpain like activity expressed in lysates from apoptotic cells, to the same extent as commercially available calpain inhibitor 1. In in vitro assays with purified calpains, indinavir and ritonavir strongly inhibit m-calpain, and moderately mu calpain. These results have great therapeutic implications, going beyond AIDS treatment, since many degenerative disorders involve abnormal calpain activation, indicating calpain as an ideal pharmacological target. Indinavir and ritonavir, potent m-calpain inhibitors, largely used since several years on humans without important negative side effects, may become powerful tools against those pathologies. PMID- 14555229 TI - S-nitrosothiol-induced rapid cytochrome c release, caspase activation and mitochondrial permeability transition in perfused heart. AB - Nitric oxide (NO) is a physiological signalling molecule, however, at high concentrations NO is cytotoxic, and has been implicated in a wide range of inflammatory, ischaemic and degenerative diseases, including heart failure. We investigated whether NO or S-nitrosothiols can induce apoptosis in perfused heart, and whether it is mediated via the mitochondrial pathway of caspase activation. We found that perfusion of rat hearts with a physiological S nitrosothiol, S-nitrosoglutathione, at 0.4-1mM concentrations for just 10 min caused the release of cytochrome c from mitochondria into the cytosol, inhibition of mitochondrial respiration and caspase activation. Inhibited mitochondrial respiration was restored when exogenous cytochrome c was added to mitochondria, indicating that respiratory inhibition was caused by lack of cytochrome c in mitochondria. Release of cytochrome c, respiratory inhibition and caspase activation were prevented when hearts were pre-perfused with cyclosporin A, suggesting that mitochondrial permeability transition pore was involved. In contrast, perfusion of the hearts with diethylenetriamine/NO adduct releasing similar levels of NO to the S-nitrosoglutathione had no measurable effect on the heart. These data suggest that S-nitrosothiols are potent inducers of apoptosis in the heart and that S-nitrosothiol-induced apoptosis is mediated by mitochondrial permeability transition but not via NO. PMID- 14555230 TI - A novel role of protein kinase C-delta in cell signaling triggered by glutathione depletion. AB - Current evidence demonstrates that protein kinase C (PKC) belongs to a group of cell-signaling molecules that are sensitive targets for redox modifications and functional alterations that mediate oxidant-induced cellular responses. Our studies have demonstrated that diminished intracellular GSH was associated to inactivation of classic isoforms and increased activity of novel PKCs, and triggered molecular signals important for cell survival. Loss of GSH and oxidative damage are probably an early signaling event in apoptotic death, which is characterized by the activation of PKC-delta. Apoptotic process consequent to GSH depletion was inhibited by rottlerin, a PKC-delta-specific inhibitor, which exerted a negative effect on oxyradical production. Therefore, it may be concluded that PKC-delta activity is related to reactive oxygen species production and is involved in the pathway leading to apoptosis and growth arrest. PMID- 14555231 TI - Chromosomal DNA fragmentation in apoptosis and necrosis induced by oxidative stress. AB - Chromosomal DNA dysfunction plays a role in mammalian cell death. Oxidative stress producing reactive oxygen species (ROS) induces chromatin dysfunction such as single- and double-strand DNA fragmentation leading to cell death through apoptosis or necrosis. More than 1 Mbp giant DNA, 200-800 or 50-300 kbp high molecular weight (HMW) DNA and internucleosomal DNA fragments are produced by oxidative stress and by some agents producing ROS during apoptosis or necrosis in several types of mammalian cells. Some nucleases involved in the chromosomal DNA fragmentation in apoptosis or necrosis are classified. ROS-mediated DNA fragmentation is caused and enhanced by polyunsaturated fatty acids (PUFAs) or their hydroperoxides through lipid peroxidation. A reduction of intracellular GSH levels induced by the inhibition of cystein transport or GSH biosynthesis leads to cell death through over production and accumulation of ROS in some types of mammalian cells. The ROS accumulation system has been used as a model of oxidative stress to discuss whether ROS-mediated DNA fragmentation associated with cell death is based on apoptosis or necrosis. PMID- 14555232 TI - The histone deacetylase inhibitor suberic bishydroxamate: a potential sensitizer of melanoma to TNF-related apoptosis-inducing ligand (TRAIL) induced apoptosis. AB - TRAIL appears to be a promising anticancer agent in that it induces apoptosis in a wide range of cancer cells but not normal tissues. Sensitivity of melanoma cells to TRAIL-induced apoptosis varied considerably because of their development of various resistance mechanisms against apoptosis. We discuss in this report the potential effect of a histone deacetylase inhibitor SBHA on TRAIL-induced apoptosis. Histone deacetylase (HDAC) inhibitors regulate histone acetylation and thereby modulate the transcriptional activity of certain genes leading to cell growth arrest, cellular differentiation, and apoptosis. Suberic bishydroxamate (SBHA) is a relatively new HDAC inhibitor that induced apoptosis in the majority of melanoma cell lines through a mitochondrial and caspase-dependent pathway. This was due to its regulation of the expression of multiple proteins that are involved in either the mitochondrial apoptotic pathway (Bcl-2 family members) or the final phase of apoptosis (caspase-3 and XIAP). Co-treatment with SBHA at nontoxic doses and TRAIL resulted in a marked increase in TRAIL-induced apoptosis of melanoma, but showed no toxicity to melanocytes. SBHA appeared to sensitize melanoma to TRAIL-induced apoptosis by up-regulation of pro-apoptotic proteins in the TRAIL-induced apoptotic pathway such as caspase-8, caspase-3, Bid, Bak, and Bax, and up-regulation of the BH3 domain only protein, Bim. This, together with activated Bid, may have acted synergistically to cause changes in mitochondria. Treatment with SBHA also resulted in down-regulation of antiapoptotic members of the Bcl-2 family, Bcl-X(L) and Mcl-1, and the IAP member, XIAP. These changes would further facilitate apoptotic signaling. SBHA appeared therefore to be a potent agent in overcoming resistance of melanoma to TRAIL-induced apoptosis. PMID- 14555233 TI - DNA damage-triggered apoptosis: critical role of DNA repair, double-strand breaks, cell proliferation and signaling. AB - Genotoxic DNA damaging agents may activate both membrane death receptors and the endogenous mitochondrial damage pathway leading to cell death via apoptosis. Here, apoptotic responses in cells exhibiting a defect in various DNA repair pathways such as alkyltransferase, base excision repair, nucleotide excision repair and mismatch repair are reviewed. The HSVTk/ganciclovir and VZV/BVDU suicide system will also be discussed. Data are available to show that critical DNA damage triggers apoptosis in a DNA replication dependent way by activating the mitochondrial damage pathway in fibroblasts. It is proposed that DNA double strand breaks (DSBs) are common ultimate apoptosis-triggering lesions arising from primary DNA lesions during DNA replication. Thus, DNA replication is a necessary component in DNA damage-triggered apoptosis, at least in fibroblasts treated with genotoxins not inducing DSBs themselves. For methylating agents inducing O(6)-methylguanine, an additional requirement is mismatch repair provoking DSB formation that triggers Bcl-2 decline and caspase-9/-3 activation. This occurs independent of p53 since most of the repair deficient cell lines under study were mutated for p53. Moreover, p53 knockout fibroblasts are more sensitive to methylating agents and UV light than p53 wt cells, suggesting p53 to play a protective rather than a pro-apoptotic role in this cell system, probably by its involvement in DNA repair. However, for lymphoblastoid cells p53 wt variants are more sensitive to DNA damage indicating that p53 participates in apoptotic signaling in a cell type-specific fashion. The role of topoisomerase II inhibitors and c-Fos/AP-1 in apoptosis will also be discussed. PMID- 14555234 TI - p73 and p63 protein stability: the way to regulate function? AB - While the p53 homologue p73 has been found to be involved in tumorigenesis, the molecular mechanisms involved in this function are still not fully evident. The presence of two distinct promoters allows the formation of two proteins with opposite effects: while TA-p73 shows pro-apoptotic effects, DeltaN-p73 has an evident anti-apoptotic function. The relative expression of the two proteins is in fact related to the prognosis of several cancers. Since both p73 and p63, the other member of the same family, share the ability to interact with each other, it is important to understand the mechanisms that control the degradation and stability of both proteins, and their relative isoforms. p73 and p63 stability is regulated not only by protein modifications (phosphorylation, acetylation) but also by its degradation in the proteasome. To this end, the interaction with Mdm2, p300/CBP, and SUMO-1 are discussed in details. PMID- 14555235 TI - Turning cell cycle controller genes into cancer drugs. A role for an antiproliferative cytokine (betaGBP). AB - Cancer therapies based on drugs designed to interfere with specific targets within the molecular circuitry of cancer cells are currently under intense experimentation. Our strategy is based on the use of a naturally occurring immunomolecule which can selectively kill cancer cells, based on its ability to exploit genetic differences between normal and cancer cells. The betaGBP cytokine has previously been shown to negatively regulate the cell cycle by blocking cells in late S phase. In tumour cells, but not in normal cells, the S phase block has been shown to be followed by apoptosis. Mechanisms involved in S phase arrest have been pinpointed to downregulation of signalling and altered expression of cell cycle controller proteins, including E2F1, a transcription factor with ability to play a part in apoptosis. Here we discuss the use of betaGBP within the context of cancer surveillance and cancer therapeutics focussing on E2F1 as one mechanistic aspect relevant to betaGBP's selective induction of programmed cell death in cancer. PMID- 14555236 TI - Apoptosis in male germ cells in response to cyclin A1-deficiency and cell cycle arrest. AB - Male mice homozygous for a mutated allele of the cyclin A1 gene (Ccna1) are sterile due to a block in cell cycle progression before the first meiotic division. Meiosis arrest in Ccna1(-/-) spermatocytes is associated with desynapsis abnormalities, lowered MPF activity, and apoptosis as evidenced by TUNEL-positive staining. With time, adult testicular tubules exhibit severe degeneration: some tubules in the older animals are almost devoid of germ cells at various stages of spermatogenesis. The mechanisms by which the cells sense the cell cycle arrest and the regulation of the decision to undergo cell death are under investigation. PMID- 14555237 TI - The multifunctional protein PEA-15 is involved in the control of apoptosis and cell cycle in astrocytes. AB - PEA-15 is a small protein (15 kDa) that was first identified as an abundant phosphoprotein in brain astrocytes [Araujo et al., J Biol Chem 1993;268(8):5911 20], and subsequently shown to be widely expressed in different tissues and highly conserved among mammals [Estelles et al., J Biol Chem 1996;271(25):14800 6; Danziger et al., J Neurochem 1995;64(3):1016-25]. It is composed of a N terminal death effector domain and a C-terminal tail of irregular structure. PEA 15 is regulated by multiple calcium-dependent phosphorylation pathways that account for its different forms: a non-phosphorylated form in equilibrium with a mono and a biphosphorylated variety. This already suggested that PEA-15 may play a major role in signal integration. Accordingly, it has been demonstrated to modulate signaling pathways that control apoptosis and cell proliferation. In particular, PEA-15 diverts astrocytes from TNFalpha-triggered apoptosis and regulates the actions of the ERK MAP kinase cascade by binding to ERK and altering its subcellular localization. The three-dimensional structure of PEA-15 has been modelized and recently determined using NMR spectroscopy, and may help to understand the various functions played by the protein through its molecular interactions. PMID- 14555238 TI - Targeting apoptosis in acute tubular injury. AB - Recent research has shown that apoptosis and its regulatory mechanisms contribute to cell number regulation in acute renal failure. Acute tubular necrosis is the most frequent form of parenchymal acute renal failure. The main causes are ischemia-reperfusion, sepsis and nephrotoxic drugs. Exogenous factors such as nephrotoxic drugs and bacterial products, and endogenous factors such as lethal cytokines promote tubular cell apoptosis. Such diverse stimuli engage intracellular death pathways that in some cases are stimulus-specific. We now review the role of apoptosis in acute renal failure, the potential molecular targets of therapeutic intervention, the therapeutic weapons to modulate the activity of these targets and the few examples of therapeutic intervention on apoptosis. PMID- 14555239 TI - Mechanisms of apoptosis in developing thymocytes as revealed by adenosine deaminase-deficient fetal thymic organ cultures. AB - Adenosine deaminase (ADA) catalyzes the conversion of adenosine and deoxyadenosine to inosine and deoxyinosine, respectively. ADA-deficient individuals suffer from severe combined immunodeficiency and are unable to produce significant numbers of mature T or B lymphocytes. This occurs as a consequence of the accumulation of ADA substrates or their metabolites. dATP is a candidate toxic metabolite because its concentration in RBCs of ADA-deficient patients correlates with the severity of disease. Murine fetal thymic organ culture (FTOC) under ADA-deficient conditions can be used as a model system to investigate the biochemical mechanism responsible for the inhibition of thymopoiesis. In ADA-deficient FTOCs initiated at day 15 of gestation, thymocyte development was arrested at the CD4(-)CD8(-)CD44(lo)CD25(+) to CD4(-)CD8( )CD44(lo)CD25(-) transition. Apoptosis appeared to be involved because the cultures could be rescued by the pan-caspase inhibitor zVADfmk, a Bcl-2 transgene, or deletion of apoptotic protease activating factor-1. As in ADA deficient patients, dATP was also elevated in ADA-deficient FTOCs. dATP levels were normalized and thymocyte development was rescued in cultures treated with an inhibitor of adenosine kinase, the enzyme that phosphorylates deoxyadenosine to dAMP. zVADfmk also prevented the accumulation of dATP in ADA-deficient FTOCs, suggesting that deoxyadenosine was derived from thymocytes undergoing apoptosis as a consequence of failing the beta selection checkpoint. In contrast, dATP levels remained elevated in ADA-deficient FTOCs with fetal thymuses from Bcl-2 transgenic mice. These data suggest that thymocyte apoptosis as a consequence of failing developmental checkpoints involves one or more caspases that are not regulated by Bcl-2. PMID- 14555240 TI - Na(+), K(+)-ATPase: the new face of an old player in pathogenesis and apoptotic/hybrid cell death. AB - The Na(+), K(+)-ATPase is a ubiquitous membrane transport protein in mammalian cells, responsible for establishing and maintaining high K(+) and low Na(+) in the cytoplasm required for normal resting membrane potentials and various cellular activities. The ionic homeostasis maintained by the Na(+), K(+)-ATPase is also critical for cell growth, differentiation, and cell survival. Although the toxic effects of blocking the Na(+), K(+)-ATPase by ouabain and other selective inhibitors have been known for years, the mechanism of action remained unclear. Recent progress in two areas has significantly advanced our understanding of the role and mechanism of Na(+), K(+)-ATPase in cell death. Along with increased recognition of apoptosis in a wide range of disease states, Na(+), K(+)-ATPase deficiency has been identified as a contributor to apoptosis and pathogenesis. More importantly, accumulating evidence now endorses a close relationship between ionic homeostasis and apoptosis, namely the regulation of apoptosis by K(+) homeostasis. Since Na(+), K(+)-ATPase is the primary system for K(+) uptake, dysfunction of the transport enzyme and resultant disruption of ionic homeostasis have been re-evaluated for their critical roles in apoptosis and apoptosis-related diseases. In this review, instead of giving a detailed description of the structure and regulation of Na(+), K(+)-ATPase, the author will focus on the most recent evidence indicating the unique role of Na(+), K(+) ATPase in cell death, including apoptosis and the newly recognized "hybrid death" of concurrent apoptosis and necrosis in the same cells. It is also hoped that discussion of some seemingly conflicting reports will inspire further debate and benefit future investigation in this important research field. PMID- 14555241 TI - Cytoskeletal reorganization during process of apoptosis induced by cytostatic drugs in K-562 and HL-60 leukemia cell lines. AB - The aim of the present study was to investigate the reorganization of F-actin, vimentin and tubulin in K-562 and HL-60 cell lines during apoptosis induced by etoposide, doxorubicin and taxol. The distribution of cytoskeletal proteins was analyzed by fluorescence microscopy. Actin was also studied by confocal microscopy and at the ultrastructural level. Changes in the distribution of cytoskeletal proteins were found to be dose-dependent and appeared to be more intense in HL-60 cells. Etoposide- and doxorubicin-treated cells showed similar changes in the distribution of F-actin, vimentin and tubulin. The reorganization of cytoskeletal proteins seemed to be consistent with features of apoptosis. An increase in bright staining of F-actin, vimentin and tubulin at the site of apoptotic bodies formation was observed. Immunogold labeling of actin in HL-60 cells was associated with features typical for apoptosis, i.e. compaction and margination of nuclear chromatin. K-562 cells showed cytoplasmic actin-positivity in the cytoplasm. Significant changes in morphology of HL-60 cells were found in the following concentrations: etoposide 20, 200 microM; doxorubicin 5, 10 microM and taxol 2-10 microM. The investigated proteins seemed to be involved in the above-reported apoptotic changes. Bright staining of F-actin, vimentin and tubulin, concentrated at the site of apoptotic bodies formation might suggested importance of these proteins for this process. Moreover, the increase in actin labeling in areas of chromatin compaction and margination of nuclear chromatin especially in HL-60 cells, which are more susceptible to apoptosis might implicate that actin might be involved in the chromatin remodeling during apoptosis. PMID- 14555242 TI - Neurofibrillary degeneration of the Alzheimer-type: an alternate pathway to neuronal apoptosis? AB - Neuronal death is a process which may be either physiological or pathological. Apoptosis and necrosis are two of these processes which are particularly studied. However, in neurodegenerative disorders, some neurons escape to these types of death and "agonize" in a process referred to as neurofibrillary degeneration. Neurofibrillary degeneration is characterized by the intraneuronal aggregation of abnormally phosphorylated microtubule-associated Tau proteins. A number of studies have reported a reactivation of the cell cycle in the neurofibrillary degeneration process. This reactivation of the cell cycle is reminiscent of the initiation of apoptosis in post-mitotic cells where G1/S markers including cyclin D1 and cdk4/6, are commonly found. However, in neurons exhibiting neurofibrillary degeneration, both G1/S and G2/M markers are found suggesting that they do not follow the classical apoptosis and an aberrant cell cycle occurs. This aberrant response leading to neurofibrillary degeneration may be triggered by the sequential combination of three partners: the complex Cdk5/p25 induces both apoptosis and the "abnormal mitotic Tau phosphorylation". These mitotic epitopes may allow for the nuclear depletion of Pin1. This latter may be responsible for escaping classical apoptosis in a subset of neurons. Since neurofibrillary degeneration is likely to be a third way to die, molecular mechanisms leading to changes in Tau phosphorylation including activation of kinases such as cdk5 or other regulators such as Pin1 could be important drug targets as they are possibly involved in early stages of neurodegeneration. PMID- 14555243 TI - Mitochondrial dysfunction, apoptotic cell death, and Alzheimer's disease. AB - Being major sources of reactive oxygen species (ROS), mitochondrial structures are exposed to high concentrations of ROS and might therefore be particularly susceptible to oxidative injury. Mitochondrial damage may play a pivotal role in the cell death decision. Bolstered evidence indicates that mitochondrial abnormalities might be part of the spectrum of chronic oxidative stress occurring in Alzheimer's disease (AD) finally contributing to synaptic failure and neuronal degeneration. Accumulation and oligomerization of amyloid beta (Abeta) is also thought to play a central role in the pathogenesis of this disease by probably directly leading to mitochondrial dysfunction. Moreover, numerous lines of findings indicate increased susceptibility to apoptotic cell death and increased oxidative damage as common features in neurons from sporadic AD patients but also from familial AD (FAD) cases. Here we provide a summary of recent work demonstrating some key abnormalities that may initiate and promote pathological events in AD. Finally, we emphasize a hypothetical sequence of the pathogenic steps linking sporadic AD, FAD, and Abeta production with mitochondrial dysfunction, caspase pathway, and neuronal loss. PMID- 14555244 TI - The essentiality of Bcl-2, PKC and proteasome-ubiquitin complex activations in the neuroprotective-antiapoptotic action of the anti-Parkinson drug, rasagiline. AB - The anti-Parkinson drug, rasagiline, a irreversible propargyl possessing monoamine oxidase B inhibitor can protect neurons in vitro and in vivo from a variety of neurotoxic insults including SIN-1, glutamate, the parkinsonism inducing neurotoxin, N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, N-methyl-(R) salsolinol and including beta amyloid protein. Recent studies have shown that rasagiline rapidly modulates intracellular signaling pathways involved in cell survival and death. Specifically rasagiline activates Bcl-2, Bcl-xl, protein kinase C (PKC) and reduces Bax in a variety of cells including PC-12 and neuroblastoma human dopamine derived SH-SY5Y cells. These enzymes play key roles in cellular events including modulation of apoptotic processes, neuronal plasticity and amyloid precursor protein processing. This pharmacological action of rasagiline is also associated with the prevention of the neurotoxin induced fall in mitochondrial membrane potential, opening of mitochondria permeability transition pore, activation of proteasome-ubiquitin complex, inhibition of cytochrome c release and prevention of caspase 3 activation, similar to the actions of cyclosporin A or Bcl-2 over expression in SH-SY5Y cells. Rasagiline and its various derivatives induces PKC dependent release of soluble amyloid precursor protein alpha and which is blocked by inhibitors of alpha-secretase, PKC and MAPK-dependent signaling. Structure-activity relationship with various propargyl containing derivatives of rasagiline including propargylamine itself has shown that the above described pharmacological action of these compounds resides in the propargylamine moiety. These results have provided a new understanding into the mechanism of neuroprotective actions of rasagiline and its anti-Alzheimer drug derivatives TV3326 and TV3279, which are relevant for therapy of Parkinson's disease, Alzheimer's disease and other neurodegenerative diseases. PMID- 14555245 TI - The mechanisms of cell death in focal cerebral ischemia highlight neuroprotective perspectives by anti-caspase therapy. AB - A number of studies have validated the importance of caspase activation in ischemia-induced brain damage. Caspases participate in both the initiation and execution phases of apoptosis, and play a central role in neuronal death after global cerebral ischemia. In focal ischemia, apoptosis occurs in the penumbra during the secondary phase of expansion of the lesion. However, ultrastructural and biochemical analysis have also shown signs of apoptosis in the initial lesion, or infarct core, which is traditionally considered necrotic. Specific caspase pathways are activated in the core and in the penumbra, and participate in both cytoplasmic and nuclear apoptotic events, notwithstanding their initial classification as activator or initiator caspases. This confirms previous suggestions that caspase inhibition holds tremendous neuroprotective potential in stroke and other apoptosis-related degenerative diseases. Consequently, two new approaches, aimed at treating stroke-induced brain damage by anti-apoptotic molecules, are being developed in academic and industrial laboratories. These are based, respectively, on the use of small peptide sequences corresponding to the preferred cleavage site of a caspase, and on genomic constructions derived from the fusion of endogenous anti-caspase molecules with a protein transduction domain from the human immunodeficiency virus-1. Fusion proteins containing endogenous caspases inhibitors efficiently counteract apoptosis in vitro. In in vivo models of focal cerebral ischemia, fusion proteins successfully cross the blood brain barrier and protect cells from ischemic death. This new approach by protein therapy could prove to be an interesting alternative for the reduction of the dramatic consequences of stroke, provided that the long-term efficiency of this protection in terms of functional recovery is demonstrated. PMID- 14555246 TI - Cell death and growth arrest in response to photodynamic therapy with membrane bound photosensitizers. AB - Photodynamic therapy (PDT) is a treatment for cancer and for certain benign conditions that is based on the use of a photosensitizer and light to produce reactive oxygen species in cells. Many of the photosensitizers currently used in PDT localize in different cell compartments such as mitochondria, lysosomes, endoplasmic reticulum and generate cell death by triggering necrosis and/or apoptosis. Efficient cell death is observed when light, oxygen and the photosensitizer are not limiting ("high dose PDT"). When one of these components is limiting ("low dose PDT"), most of the cells do not immediately undergo apoptosis or necrosis but are growth arrested with several transduction pathways activated. This commentary will review the mechanism of apoptosis and growth arrest mediated by two important PDT agents, i.e. pyropheophorbide and hypericin. PMID- 14555248 TI - Is the paradigm shifting? AB - Reduction in bone mass has long dominated the thinking about and approach to the problem of osteoporosis. A now large body of evidence indicates that bone mass is not adequate to explain satisfactorily either the skeletal fragility of osteoporosis or the effects of bone active agents. By contrast, bone remodeling activity seems to provide a better explanation of both. It is suggested that current syntheses in the field are shifting to this conclusion. In attempting to make sense out of how a process designed by evolution to sustain bone strength could instead be contributing to weakness, I suggest: (1) prevailing bone remodeling levels are substantially higher than are optimal for maintenance of bone strength; (2) this high level has discernible environmental causes; and (3) such high remodeling is a major source-perhaps the major source-of osteoporotic bony fragility. Within this context, reduced bone mass, rather than the primary cause of fracture, is seen as a factor that predisposes individuals to the harmful effects of excessive remodeling. PMID- 14555249 TI - The lesson of temporary brittle bone disease: all bones are not created equal. AB - Temporary brittle bone disease (TBBD) is a recently described phenotype of multiple, unexplained fractures in the first year of life and predominantly in the first 6 months of life. There is usually no other injury such as bruising, subdural hematomas, retinal hemorrhages, or other internal organ injury. The susceptibility to fracture is transient, and there are no other radiographic or biochemical abnormalities noted in the standard evaluation that might suggest an underlying cause. The child abuse and pediatric radiology communities have, for the most part, been unwilling to accept this as a real condition, for they believe it is a ruse for child abuse. This review describes the experience of the author in evaluating infants with multiple unexplained fractures and the hypothesis that has emerged for explaining TBBD. The hypothesis is a prenatal application of the mechanostat/bone loading theory of bone formation and states that TBBD is caused by fetal immobilization which leads to fetal bone unloading and transient, relative osteopenia. Such susceptible infants can fracture with routine handling and present with a pattern of fractures that is similar to that which has been thought to be highly specific for child abuse. The review presents: (a) the evidence that indicates that normal fetal movement is important for normal fetal bone strength, (b) a critique of the radiologic approach in the diagnosis of child abuse in infants with multiple unexplained fractures, (c) observations that would indicate that child abuse is unlikely in infants with TBBD, and (d) new approaches to the infant with multiple unexplained fractures that would assist in accurate diagnosis. PMID- 14555247 TI - Connexins and apoptotic transformation. AB - We examined the influence of connexin (Cx) expression on the development of apoptosis in HeLa parental cells (coupling deficient cell line) and HeLa cells expressing wild-type Cx43 and Cxs fused with enhanced green fluorescent protein (EGFP). EGFP was attached to the C-terminus of Cx32 and Cx43, Cx32-EGFP and Cx43 EGFP, respectively, and to the N-terminus of Cx32, EGFP-Cx32. All fusion proteins assembled into junctional plaques (JPs) at areas of cell-cell contact, but only the C-terminal fusion proteins formed functional gap junction (GJ) channels as well as hemichannels. In each cell line, apoptosis was induced by treatment with various agents including anisomycin, camptothecin, cis-platinum, colchicine, cycloheximide, etoposide, staurosporin and taxol. Using fluorescence microscopy, time-lapse imaging and dual whole-cell voltage clamp techniques, we correlated the changes in functional properties of GJ channels and Cx distribution with the progression of apoptosis based on cells' labeling with acridine orange and ethidium bromide (EB). The early phase of apoptosis (a viable apoptotic (VA) state) was characterized by shrinkage of the cells and by increased internalization of JPs accompanied by decreased cell-cell coupling. The apoptotic reagents had no direct effect on electrical cell-cell coupling. Transformation from a VA to a nonviable apoptotic (NVA) state was faster in HeLa cells expressing Cx43 or Cx43-EGFP than in HeLa parental cells. The potent GJ uncoupler, octanol, slowed the transition of HelaCx43-EGFP cells into a NVA state. In the absence of apoptotic reagents, the rate of EB uptake was higher in HeLaCx43-EGFP than in HeLa parental cells consistent with the presence of open Cx43-EGFP hemichannels. However, in both cell lines the rate of EB uptake decreased proportionally during the development of apoptosis suggesting that membrane permeability ascribed to Cx hemichannels is reduced. Cells expressing Cx32-EGFP and EGFP-Cx32 demonstrate the same apoptotic patterns as HeLaCx43-EGFP and HeLa parental cells, respectively. Intracellular levels of ATP in HeLaCx43 EGFP cells were substantially lower than in HeLa parental cells, and ATP added to the medium abolished the accelerated transition from a VA to a NVA state in HeLaCx43-EGFP cells. In summary, Cx32 or Cx43 accelerates transformation of cells into a NVA state or secondary necrosis and this depends on the ability of Cxs to form functional GJ channels and hemichannels. PMID- 14555250 TI - Uniaxial cyclic stretch induces osteogenic differentiation and synthesis of bone morphogenetic proteins of spinal ligament cells derived from patients with ossification of the posterior longitudinal ligaments. AB - Ossification of the posterior longitudinal ligament of the spine (OPLL) is characterized by ectopic bone formation in the spinal ligaments. Mechanical stress, which acts on the posterior ligaments, is thought to be an important factor in the progression of OPLL. To elucidate this mechanism, we investigated the effects of in vitro sinusoidal cyclic stretch (120% peak to peak, at 1 Hz) on cultured spinal ligament cells derived from OPLL and non-OPLL patients. The mRNA expressions of alkaline phosphatase (ALP), osteopontin, bone morphogenetic protein (BMP)-2, BMP-4, and BMP receptors as well as ALP activity in cell layers and production of BMPs into the conditioned medium were significantly increased by cyclic stretch in OPLL cells, whereas no change was observed in non-OPLL cells. A stretch-activated Ca(2+) channel blocker, Gd(3+), the voltage-dependent L-type Ca(2+) channel blockers diltiazem and nifedipine, and Ca(2+)-free medium suppressed stretch-induced ALP activity, which suggests a role of Ca(2+) influx in the signal transduction of mechanical stress to the osteogenic response of OPLL cells. Our study provides first evidences that mechanical stress plays a key role in the progression of OPLL through the induction of osteogenic differentiation in spinal ligament cells and the promotion of the autocrine/paracrine mechanism of BMPs in this lesion. PMID- 14555251 TI - Effects of endurance exercise on three-dimensional trabecular bone microarchitecture in young growing rats. AB - Appropriate endurance exercise is capable of increasing bone mass and strength in both animals and humans. We examined the skeletal changes induced by treadmill running exercise in young growing rats with a particular emphasis on three dimensional trabecular bone microarchitecture. Fourteen male Wistar rats were divided into sedentary (CON; n = 7) and exercised (RUN; n = 7) groups at the age of 4 weeks. The rats in the RUN group performed the treadmill running exercise of 30 m/min for 60 min, 5 times a week. After 10 weeks of exercise, bone mineral density (BMD), cortical geometry, diaphyseal breaking force, and trabecular bone microarchitecture in the femur were measured. Three-dimensional trabecular bone microarchitecture was evaluated at the distal femoral metaphysis using microcomputed tomography. The running exercise significantly increased BMD, bone volume, bone volume fraction, trabecular thickness, and trabecular number, whereas trabecular bone pattern factor, the parameter associated with decreased trabecular connectivity, was significantly lower in the RUN group than the CON group. On the other hand, no significant difference in the degree of anisotropy and structure model index was observed between the two groups. At the femoral diaphysis, running exercise significantly increased cortical bone area, width, and maximum load without affecting bending stress, implying that the material properties of bone had not changed in the exercised rats. These results suggest that the increase in bone strength induced by endurance exercise is mediated by changes in trabecular bone microarchitecture as well as density and cortical geometry. PMID- 14555252 TI - On the importance of geometric nonlinearity in finite-element simulations of trabecular bone failure. AB - The finite element method, which has been successfully applied to studies of the elastic properties of trabecular bone, is now being used to simulate its failure. These simulations have used a geometrically linear (linear kinematic) approximation to the total stiffness matrix; nonlinear terms in the total stiffness matrix have been excluded from the computation in order to achieve efficiency. Because trabecular bone appears to be a slender (i.e., geometrically nonlinear) structure, we studied the validity of the linear kinematic approximation for simulating its failure. Two cases, designed to bracket the extremes of stability behavior, were explored: a single representative spicule of trabecular bone (case 1) and a volume of trabecular bone consisting of relatively low aspect ratio members (case 2). For case 1, geometrically linear (GL) and nonlinear (GNL) analyses were performed with two different materials models: a plastic damage model and a brittle damage model. When GNL terms were included in the total stiffness matrix, we found that load-path bifurcation preceded tissue failure regardless of the form of the damage model. This bifurcation was the result of a complex coupling between material yield and structural instability. The nature of this coupling was highly sensitive to the form of the damage model. None of these behaviors was observed in the linear analyses, where failure was insensitive to the form of the damage model and where structural instabilities were prevented from occurring. For case 2, compressive loading of a volume of trabecular bone, geometric nonlinear effects were pronounced. There was a bifurcation in load response that resulted in large apparent strain to failure. The GL simulations, on the other hand, precluded this bifurcation. We hypothesize that trabecular bone is a geometric nonlinear structure; nonlinear terms must be included in the total stiffness matrix to accurately simulate its failure. PMID- 14555253 TI - Characteristics of a prevalent vertebral deformity predict subsequent vertebral fracture: results from the European Prospective Osteoporosis Study (EPOS). AB - The presence of a prevalent vertebral deformity increases the risk of a future vertebral fracture. The aim of this study was to determine whether certain characteristics of the prevalent deformity, including its shape and location in the spine, influenced this effect. The 3100 men and 3500 women who took part in this analysis were recruited from population registers for participation in the European Prospective Osteoporosis Study (EPOS). Subjects had lateral thoracic and lumbar spine x-rays at baseline, and again after a mean interval of 3.8 years. Prevalent morphometric vertebral deformities on the baseline film were identified by the McCloskey-Kanis method. Incident fractures were defined as vertebrae that also satisfied the McCloskey-Kanis criterion for prevalent deformities on the follow-up film, and in addition had at least one height (anterior, mid, or posterior) which had reduced by at least 20% between films. Poisson regression was used to assess the association between various characteristics of the prevalent deformity and the risk of an incident vertebral fracture, with generalised estimating equations used to allow for the fact that each subject contributed several vertebrae to the analysis. The risk of an incident fracture increased with the number of prevalent deformities: relative risk (RR) for one prevalent deformity 3.2 (95% confidence interval (CI); 2.1, 4.8); 9.8 (95% CI;6.1, 15.8) for 2; and 23.3 (95% CI;15.3, 35.4) for 3 or more. Relative risks differed significantly according to the shape of the prevalent deformity, ranging from 5.9 (95% CI; 4.1, 8.6) if the anterior and mid heights were reduced to 1.6 (95% CI;0.8, 3.2) if the posterior and mid heights were reduced. Risks varied also according to the severity of the deformity. There were fivefold differences in relative risk of incident fracture depending on the location of the prevalent deformity within the spine. Compared to vertebrae in subjects with no deformities at baseline, the relative risk of an incident fracture within three vertebrae of a prevalent deformity was greater (7.7 (95% CI;5.6, 10.5)) than the risk in more distant vertebrae (4.0 (95% CI;2.6, 6.0)). In summary, the risk of a subsequent vertebral fracture in individuals with preexisting deformities is importantly influenced by the characteristics of these deformities. PMID- 14555254 TI - In situ chemistry of osteoporosis revealed by synchrotron infrared microspectroscopy. AB - Reduced bone density is a well-known feature of osteoporosis, yet little is known about the changes in the chemical composition of bone or the impact of such chemical changes on fracture risks. Using ovariectomized cynomolgus monkeys (Macaca fascicularis) as a model for the menopausal onset of osteoporosis, we examined the microscopic chemical changes of bone measured by synchrotron infrared microspectroscopy as a function of time after ovariectomy. The results demonstrate that cortical bone formed 1 or 2 years after ovariectomy, as identified by fluorochrome labeling, has a higher phosphate content (PO4(3 )/matrix ratio), a lower carbonate content (CO3(2-)/matrix ratio), and more mature collagen cross-links (nonreducible cross-link/reducible cross-link ratio) than that formed in sham-operated controls. Trabecular bone after ovariectomy shows no changes in phosphate content, a lower carbonate content, and immature collagen cross-linking. Treatment with a bone turnover suppressor, (nandrolone decanoate) reverses most of the ovariectomy-induced chemical changes in the cortical bone to the levels of the ovary-intact controls, but has little effect on the trabecular bone. These results demonstrate that bone newly synthesized after ovariectomy is chemically different from healthy bone within specific bone regions, which may contribute to reduced bone quality in osteoporosis. PMID- 14555255 TI - Severity of prevalent vertebral fractures and the risk of subsequent vertebral and nonvertebral fractures: results from the MORE trial. AB - Prevalent vertebral fractures and baseline bone mineral density (BMD) predict subsequent fracture risk. The objective of this analysis is to examine whether baseline vertebral fracture severity can predict new vertebral and nonvertebral fracture risk. In the randomized, double-blind 3-year Multiple Outcomes of Raloxifene Evaluation (MORE) trial, 7705 postmenopausal women with osteoporosis (low BMD or prevalent vertebral fractures) were randomly assigned to placebo, raloxifene 60 mg/day, or raloxifene 120 mg/day. Post hoc analyses studied the association between baseline fracture severity and new fracture risk in the placebo group and the effects of placebo, raloxifene 60 mg/day, and raloxifene 120 mg/day on new fracture risk in women with the most severe prevalent vertebral fractures (n = 614). Vertebral fracture severity was visually assessed using semiquantitative analysis of radiographs and categorized by estimated decreases in vertebral heights. Reported new nonvertebral fractures were radiographically confirmed. Baseline vertebral fracture severity predicted vertebral and nonvertebral fracture risk at 3 years. In women without prevalent vertebral fractures, 4.3 and 5.5% had new vertebral and nonvertebral fractures, respectively. In women with mild, moderate, and severe prevalent vertebral fractures, 10.5, 23.6, and 38.1% respectively had new vertebral fractures, whereas 7.2, 7.7, and 13.8% respectively experienced new nonvertebral fractures. Number of prevalent vertebral fractures and baseline BMD also predicted vertebral fracture risk, but the severity of prevalent vertebral fractures was the only predictor of nonvertebral fracture risk and remained a significant predictor after adjustment for baseline characteristics, including baseline BMD. In patients with severe baseline vertebral fractures, raloxifene 60 mg/day decreased the risks of new vertebral [RR 0.74 (95% Cl 0.54, 0.99); P = 0.048] and nonvertebral (clavicle, humerus, wrist, pelvis, hip, and leg) fractures [RH 0.53 (95% CI 0.29, 0.99); P = 0.046] at 3 years. To prevent one new fracture at 3 years in women with severe baseline vertebral fractures with raloxifene 60 mg/day, the number needed to treat (NNT) was 10 for vertebral and 18 for nonvertebral fractures. Similar results were observed in women receiving raloxifene 120 mg/day. In summary, baseline vertebral fracture severity was the best independent predictor for new vertebral and nonvertebral fracture risk. Raloxifene decreased new vertebral and nonvertebral fracture risk in the subgroup of women with severe vertebral fractures at baseline. These fractures may reflect architectural deterioration, independent of BMD, leading to increased skeletal fragility. PMID- 14555256 TI - Bone response to jumping is site-specific in children: a randomized trial. AB - Skeletal loading during growth may be one way of increasing bone mass early in life. We hypothesized that children randomized to a jumping program (25 jumps/day from a 45-cm box, 5 days/week for 12 weeks) would have greater increases in hip and spine bone mineral content (BMC) and 4% distal tibia volumetric bone density than children randomized to the control group. Our secondary hypothesis was that jumping would not be as beneficial among peripubertal children as compared to prepubertal or pubertal children due to the relatively high growth rate that occurs during the peripubertal period. Fifty-four children (31 girls) ages 3-5, 7 8, 11-12, and 15-18 years were enrolled. We performed bone, anthropometric, and force plate measurements at baseline and 12 weeks. Twenty-four-hour diet recall and Tanner's self-report of pubertal development were completed at baseline. Jumpers had a lower calcium intake than nonjumpers at baseline (965 +/- 403 vs 1295 +/- 465 mg/day, P < 0.01), but the groups were otherwise similar. Overall, jumpers had greater increases in total body BMC (45.0 +/- 4.9 vs 29.4 +/- 5.3 g, P = 0.03) and regional dual energy x-ray absorptiometry leg BMC (19.8 +/- 2.6 vs 11.5 +/- 2.8 g, P = 0.03) than nonjumpers at all pubertal stages. However, the 4% distal tibia bone response to jumping appeared to be modified by pubertal stage, with the greatest bone benefit from jumping observed in pubertal children (interaction of jumping group by pubertal stage, P < 0.05, for both BMC and volumetric BMD). A similar pattern was observed for spine BMC (interaction, P = 0.10). We conclude that skeletal loading increases total body and leg BMC in children, but may not have a positive effect at sites that are predominantly trabecular bone during periods of rapid growth (i.e., peripubertal period). PMID- 14555257 TI - Bone density in sheep: genetic variation and quantitative trait loci localisation. AB - Bone density (BD) is an important factor in osteoporotic fracture risk in humans. However, BD is a complex trait confounded by environmental influences and polygenic inheritance. Sheep provide a potentially useful model for studying differences in BD, as they provide a means of circumventing complex environmental factors and are a similar weight to humans. The aims of this study were to establish whether there is genetic variation in BD in sheep and then to localise quantitative trait loci (QTLs) associated with this variation. We also aimed to evaluate the relationship between fat and muscle body components and BD in sheep. Results showed that there was significant (P < 0.01) genetic variation among Coopworth sheep sires for BD. This genetic difference was correlated (P < 0.01) with body weight and muscle mass. A number of QTLs exceeding the suggestive threshold were identified (nine in total). Of these, two (chromosomes 1, P < 0.05; chromosome 24, P < 0.01) were significant using genome-wide permutation significance thresholds (2000 iterations). The position of the QTL on chromosome 24 coincided with a number of other body composition QTLs, indicating possible pleiotropic effects or the presence of multiple genes affecting body composition at that site. This study shows that sheep are potentially a useful model for studying the genetics of BD. PMID- 14555258 TI - The effect of enterocystoplasty on bone strength assessed at four different skeletal sites in a rat model. AB - The objective of the study was to investigate bone strength at four different skeletal sites in a chronic animal model of urinary diversion. Young male Wistar rats (120) were allocated randomly to four groups undergoing ileocystoplasty; ileocystoplasty and resection of the ileocecal segment; colocystoplasty; or sham operation (controls). After 8 months the lumbar vertebrae, femora, and tibiae were harvested at necropsy. Bone strength was assessed biomechanically at four different skeletal sites: vertebra L3, femoral middiaphysis, femoral neck, and distal femoral metaphysis. Bone mass and architecture were assessed using standard static histomorphometry of the proximal tibial metaphysis (trabecular bone volume [BV/TV]; trabecular number [Tb.N]) and ash weight. Statistically significant differences of biomechanical parameters between groups were observed at three skeletal sites with corresponding changes in tibial histomorphometry. Isolated ileocystoplasty resulted in decreased maximum load values of L3 (-16.4%; p < 0.0035) and a substantial reduction in tibial BV/TV (-34.7%; p < 0.05). Ileocystoplasty combined with resection of the ileocecal segment led to a significant loss of bone strength of L3 (-32.4%; p < 0.0015) and a dramatic reduction of tibial BV/TV (-45.9%; p < 0.01). Loss of tibial metaphyseal bone mass was predominantly caused by a decrease in Tb.N. (p < 0.01). Colonic augmentation had no significant effect on bone strength or histomorphometric values. In conclusion, this is the first experimental study to demonstrate the relevance of histomorphometrically proven bone loss after enterocystoplasty in terms of biomechanical variables. PMID- 14555260 TI - Long-term analgesic effect of clodronate in rodents. AB - Several studies have shown that treatment with bisphosphonates can reduce the pain associated with different painful diseases. In a previous study we demonstrated that in mice two bisphosponates, clodronate and pamidronate, had an antinociceptive effect under acute conditions not related to bone processes, after in vein (iv) or intracerebroventricular (icv) injection. The present study tested the time-dependent antinociceptive action of clodronate and pamidronate in comparison with that of acetylsalicylic acid (ASA) and morphine after iv and icv injection using the tail-flick test in acute and chronic treatment. The effects of clodronate on other measures of animal behaviour were also evaluated. In the tail-flick test, administration of clodronate iv produced an antinociceptive effect that was greater than that of ASA and statistically significant up to 16 h; pamidronate iv showed a significant antinociceptive effect for only 6 h. Clodronate and pamidronate icv showed an increase in tail-flick latency time that was significant and lasted for 16 and 6 h, respectively, while morphine produced an antinociceptive effect for 24 h. In the test we found significant differences between male and female mice in the latency time values but not in the length of the analgesic effect. In the chronic treatment paradigm, clodronate produced a significant increase of the tail-flick latency after the first injection. The analgesic effect increased up to 50% after 5 days of treatment. Significant analgesic effects were still present after 3, 7, and 14 days from the end of treatment. Clodronate did not produce any significant behavioural effects in the Rota-rod test, pentobarbital-induced sleeping time, and locomotor activity cage. These data indicate that clodronate presents a central and peripheral prolonged antinociceptive effect, without any behavioural side effects. PMID- 14555259 TI - Comparative effects of vitamin K and vitamin D supplementation on prevention of osteopenia in calcium-deficient young rats. AB - The aim of this study was to clarify the difference in the effects of vitamin K and vitamin D supplementation on the development of osteopenia in young rats under mild calcium deficiency. Sixty female Sprague-Dawley rats, 6 weeks of age, were randomized by stratified weight method into six groups with 10 rats in each group: baseline control, 0.5% (normal) calcium diet, 0.1% (low) calcium diet, 0.1% calcium diet + vitamin K (30 mg/100 g, food intake), 0.1% calcium diet + vitamin D (25 microg/100 g, food intake), and 0.1% calcium diet + K + D. After 10 weeks of feeding, serum calcium, 25-hydroxyvitamin D(3) [25 (OH) D(3)], 1,25 dihydroxyvitamin D(3) [1,25 (OH)(2) D(3)], and parathyroid hormone (PTH) levels were measured, and intestinal calcium absorption and renal calcium reabsorption were evaluated. Bone histomorphometric analyses were performed on cortical bone of the tibial shaft and cancellous bone of the proximal tibia. Calcium deficiency induced hypocalcemia, increased serum PTH and 1,25 (OH)(2) D(3) levels with decreased serum 25 (OH) D(3) level, stimulated intestinal calcium absorption and renal calcium reabsorption, and reduced maturation-related cortical bone gain as a result of decreased periosteal bone gain and enlarged marrow cavity but did not significantly influence maturation-related cancellous bone gain. Vitamin K supplementation in calcium-deficient rats stimulated renal calcium reabsorption, retarded the abnormal elevation of serum PTH level, increased maturation-related cancellous bone gain, and retarded the reduction in maturation-related cortical bone gain. On the other hand, vitamin D supplementation in calcium-deficient rats stimulated intestinal calcium absorption via increased serum 1,25 (OH)(2) D(3) level with prevention of the abnormal elevation of serum PTH level, prevented hypocalcemia, reduced the maturation-related cancellous bone gain, and prevented the reduction in periosteal bone gain and enhanced enlargement of the marrow cavity with no significant effect on the reduction in maturation-related cortical bone gain. However, no synergistic effect of vitamin K and vitamin D on intestinal calcium absorption, renal calcium reabsorption, and cancellous and cortical bone mass was found. This study shows the differential effects of vitamin K and vitamin D supplementation on the development of osteopenia in young rats under mild calcium deficiency. Vitamin K supplementation stimulates renal calcium reabsorption, increases maturation-related cancellous bone gain, and retards the reduction in maturation-related cortical bone gain, whereas vitamin D supplementation stimulates intestinal calcium absorption and prevents the reduction in maturation-related periosteal bone gain by inducing accumulation of calcium from cancellous and endocortical bone. PMID- 14555261 TI - Effects of 4-year treatment with once-weekly clodronate on prevention of corticosteroid-induced bone loss and fractures in patients with arthritis: evaluation with dual-energy X-ray absorptiometry and quantitative ultrasound. AB - The aim of this placebo-controlled study was to determine whether once-weekly clodronate could prevent osteoporosis in patients with arthritis at the start of corticosteroid therapy. One hundred sixty-three patients, 18 to 90 years of age, with rheumatoid or psoriatic arthritis, were randomly assigned to receive either clodronate (100 mg im/week) plus calcium and vitamin D (1000 mg and 800 UI, respectively) or calcium and vitamin D alone. Patients had started therapy with prednisone or its equivalent within the previous 100 days and had bone mineral density <2.5 SD below mean young normal values at the lumbar spine or femoral neck. The primary outcome was the difference between the two treatment groups at months 12, 24, 36, and 48 in the mean percentage change from baseline in the bone mineral density of the lumbar spine, femur (neck and total), and total body. Secondary measurements included changes in the stiffness index evaluated by ultrasound measurements and the rate of new vertebral fractures. The bone density and stiffness did not change significantly in the clodronate plus calcium and vitamin D group, whereas it declined significantly in the calcium plus vitamin D group. The difference between treatment groups at 48 months in the mean change from baseline was 8.78 +/- 1.4% for the lumbar spine (P < 0.01), 7.31 +/- 1.12% for the femoral neck (P < 0.01), 7.92 +/- 1.93% for the trochanter (P < 0.01), 8.39 +/- 1.80% for total femur (P < 0.01), 6.94 +/- 1.09% for total body (P < 0.01), and 9.38 +/- 2.21% for stiffness of os calcis (P < 0.01). Depending on the skeletal regions evaluated, 85 to 98% of patients treated with clodronate had a densitometric change lower than the lowest significant densitometric difference. One hundred percent of patients treated with calcium plus vitamin D had a densitometric decrease greater than the lowest significant difference. The relative risk of vertebral fractures and multiple vertebral fractures in the clodronate group compared to the calcium plus vitamin D group was 0.63 (0.35 0.98, 95% CI) and 0.25 (0.15-0.91, 95% CI), respectively. We concluded that pulsatory administration of im clodronate once weekly is a safe therapy for preventing corticosteroid induced osteoporosis in patients with arthritis. PMID- 14555262 TI - Effect of intravenous pamidronate on bone markers and local bone mineral density in fibrous dysplasia. AB - Bisphosphonates have proven to be effective in patients with fibrous dysplasia of the bone (FD) as shown by their effect on bone pain, markers of bone turnover, or radiological changes. The aim of this study was to evaluate the usefulness of measuring bone mineral density (BMD) of affected bones to assess the efficacy of bisphosphonate treatment. Seven patients (mean age 26 years) received courses of 180 mg intravenous infusion of pamidronate every 6 months (60 mg/day during 3 days). Clinical symptoms, serum alkaline phosphatase, and urinary C-terminal cross-linking telopeptide of type I collagen were assessed every 3 months. BMD of total skeleton and X-rays of FD areas (FDa) were performed at baseline and at 12 months. BMD of FDa was compared with the contralateral side (CL) using the region of interest program on the total skeleton scan. BMD of total skeleton was normal at baseline. Average BMD of FDa was -11.4% compared with CL, a significantly greater difference than that observed between the left and right sides in healthy controls, -0.7% (P < 0.02). At 12 months bone pain diminished in all patients. Bone turnover markers decreased. Mean total skeleton BMD increased 3.3% (P < 0.02). Subregions of the total skeleton scan presenting FD lesions augmented: arms +9.6% (P < 0.02), legs +4.2%, and pelvis +3.5% (P < 0.05). The increase in mean BMD of FDa was +6.8% compared with +2.6% in CL. No changes were observed on the X-ray. These results indicate that simultaneous determination of markers of bone turnover and BMD of FDa is useful in short-term follow-up to determine the efficacy of intravenous pamidronate. PMID- 14555263 TI - The effects of three-month intravenous ibandronate on bone mineral density and bone remodeling in Klinefelter's syndrome: the influence of vitamin D deficiency and hormonal status. AB - The aim of this study was to evaluate the effects of a 2-year treatment with intravenous ibandronate (2 mg every 3 months) and calcium (1000 mg daily) on bone mineral density (BMD) and bone markers in 14 patients with Klinefelter's syndrome who served as their own controls. During the follow-up of 5.9 years before the treatment was started, the mean rates of bone loss per year were 1.3, 0.9, and 0.6% in the lumbar spine, femoral neck, and total body, respectively. The rate of bone loss from the spine was significantly inversely related to both serum estradiol and testosterone. At the onset of treatment, the average age of the patients was 55.2 years (48-64 years), and T score, mean +/- SD, at the lumbar spine was -2.6 +/- 1.0. After 6 months, the mean serum CTX and PINP decreased by 39 and 55% below the pretreatment concentrations, respectively (P < 0.05). After 12 months of treatment, the patients gained mean +/- SD, 7.8 +/- 2.3% of BMD in the lumbar spine, 3.8 +/- 4.0% in the femoral neck, and 4.7 +/- 2.2% in the total body (P < 0.05). During the second year of treatment, all patients also received 700 IU of vitamin D daily. After 24 months of treatment, the patients gained 10.1 +/- 4.3% of BMD in the lumbar spine, 6.7 +/- 5.5% in the femoral neck, and 5.5 +/ 2.5% in the total body. The increase in BMD in the second year of ibandronate treatment was not significant. The rate of gain of BMD in the femoral neck was positively related to serum concentrations of testosterone and inversely related to 25-hydroxyvitamin D (P < 0.005). After the discontinuation of treatment, serum CTX and PINP increased to the pretreatment levels, and the lumbar spine and femur neck BMD decreased (P < 0.05). In conclusion, ibandronate was effective in increasing BMD at all sites, but the effects were adversely influenced by vitamin D insufficiency or deficiency. The overall changes in biochemical markers of bone remodeling were consistent with the antiresorptive effect of the drug. PMID- 14555264 TI - Isoflavones with supplemental calcium provide greater protection against the loss of bone mass and strength after ovariectomy compared to isoflavones alone. AB - Although hormone replacement therapy (HRT) and calcium (Ca) supplementation preserve bone mass more when combined, there is a growing concern over the safety of HRT that necessitates thorough investigation of effective, alternative treatments for bone loss. While plant-derived estrogen-like compounds such as isoflavones preserve bone, it is not known whether isoflavones and Ca supplementation attenuate losses in bone mass and strength to a greater extent when combined. This study compared the effects of an isoflavone extract + high Ca to isoflavone extract or high Ca alone on preservation of bone mineral density (BMD) and biomechanical strength in ovariectomized (ovx) rats. Rats were sham operated (n = 10) or ovx (n = 40). Shams were fed a 0.2% Ca diet. Ovx rats were randomized to a 0.2% Ca diet alone (OVX) or with isoflavone extract (IE; 1.6 g/kg diet) or to a high Ca diet (Ca; 2.5%) alone or a high Ca diet with the isoflavone extract (IE + Ca) for 8 weeks. BMD of femur and lumbar spine were measured by dual-energy X-ray absorptiometry. The biomechanical strength of femurs and individual vertebra was measured by three-point bending and compression testing, respectively. The average food intake was lowest (P < 0.05) among sham and IE groups and greatest (P < 0.05) among the OVX group. Final body weight was lowest (P < 0.05) among shams and highest (P < 0.05) among the OVX group while IE + Ca were lighter (P < 0.05) than all ovx groups. Femur and vertebra BMD was greater (P < 0.05) among IE + Ca and sham rats compared to IE, Ca, or OVX rats. Although there were differences in femur BMD among groups, biomechanical properties at the femur midpoint did not differ among groups, possibly due to the lack of cortical bone loss at this site. Conversely, vertebra biomechanical strength was greater (P < 0.05) among IE + Ca and Ca alone groups compared to IE alone. Uterine weight was higher (P < 0.05) among shams than OVX and IE with no difference among shams, Ca, or IE + Ca rats, suggesting that the isoflavones did not have an uterotrophic effect. In conclusion, isoflavones combined with high Ca are more protective against the loss of femur and vertebra BMD than isoflavones or high Ca diet alone. PMID- 14555266 TI - Tests of linkage and association of the COL1A2 gene with bone phenotypes' variation in Chinese nuclear families. AB - In the present study, we simultaneously test linkage and/or association of the collagen type I alpha 2 (COL1A2) gene with bone mineral density (BMD) and bone area. A total of 1280 subjects from 407 Chinese nuclear families (including both parents and their daughters) were genotyped for an intragenic marker MspI in the COL1A2 gene. BMD and bone area at the lumbar spine and hip were measured by dual energy X-ray absorptiometry. Applying the QTDT (quantitative transmission disequilibrium test) program, we performed tests for population stratification, within-family association (via transmission disequilibrium test), total association, linkage, and linkage while modeling association. Significant or marginal within-family associations were found with BMD at the lumbar spine (P = 0.013), trochanter (P = 0.004), and total hip (P = 0.053) and with bone area at the intertrochanteric region (P = 0.024) and total hip (P = 0.048). The positive associations were confirmed in permutations except for bone area at total hip (P > 0.10). A small proportion (<1%) of the population variance of bone phenotypes can be explained by the MspI polymorphism; however, it may be underestimated given the significant population stratification detected in our sample. Due to the limited number of sib pairs in this sample, we did not find evidence of linkage. In summary, the MspI polymorphism is likely to be in linkage disequilibrium with a nearby functional mutation affecting BMD and bone area. PMID- 14555265 TI - Interrelationship between bone turnover markers and dietary calcium intake in pregnant women: a longitudinal study. AB - This longitudinal study evaluated bone turnover and the interrelationship between changes in bone biomarkers and habitual dietary calcium intake during pregnancy in a group of women ranging widely with regard to dietary calcium intake. Thirty nine healthy pregnant and 30 nonpregnant women were studied. Calcium, phosphorus, 1alpha,25-dihydroxyvitamin D (1,25diHOD), bone alkaline phosphatase (bALP), carboxyterminal propeptides of type I procollagen (PICP) and carboxyterminal telopeptides of type I collagen (betaCTX and ICTP) were measured in serum and calcium, and creatinine and aminoterminal telopeptide (NTX) were determined in urine. Serum calcium and phosphorus did not change but the urinary Ca/Creat ratio and 1,25diHOD increased throughout pregnancy (P < 0.001 and P < 0.0001, respectively). Serum b-ALP and PICP increased during the last two trimesters (P < 0.0001 and P < 0.001, respectively). All studied bone resorption markers increased compared to nonpregnant values throughout pregnancy. The highest increment was observed in the third trimester. The level of significance decreased as follows: betaCTX > NTX >ICTP. Serum 1,25 diHOD versus calcium intake showed a positive and significant correlation (r = 0.51, P < 0.02). A negative correlation between the absolute change in betaCTX, NTX, and b-ALP between the third and second trimester and calcium intake at the end of pregnancy was observed in pregnant women who did not cover adequately calcium intake requirements (r = -0.47, P < 0.03; r = -0.41, P < 0.05; and r = -0.43, P < 0.05, respectively). These results suggest that skeletal response to pregnancy may not be entirely independent of maternal calcium intake, especially in women with usually low calcium intake. In summary, not only hormonal changes in calcium metabolism that occur during pregnancy but also other considerations, such as low dietary calcium intake, may lead to an increment in the biological activity of the skeleton. Additional studies must be conducted to confirm our findings and to gain a better understanding of skeletal response to a low calcium intake during pregnancy. PMID- 14555267 TI - Older people in China and the United Kingdom differ in the relationships among parathyroid hormone, vitamin D, and bone mineral status. AB - To explore whether there are ethnic differences in relationships among parathyroid hormone (PTH), vitamin D, and bone mineral status, 352 healthy volunteers, 60-83 years old, were studied in Shenyang, Peoples' Republic of China (108 men, 110 women), and in Cambridge, UK (67 men, 67 women), in late winter. Early morning fasting blood and 2-h fasting urine were analyzed for 25 hydroxyvitamin D (25OH-D), PTH, and free deoxypyridinoline (DPD). Hip bone mineral status was measured using dual-energy X-ray absorptiometry (Lunar). There were significant differences (P < 0.001) in plasma 25OH-D and PTH concentrations between Shenyang and Cambridge [25OH-D nmol/L: Shenyang = 29.0 (SD 12.7), Cambridge = 35.7 (12.9)]; PTH ng/L: Shenyang = 34.3 (13.4), Cambridge = 25.2 (11.0)]. PTH was negatively related to 25OH-D in both populations. The relationship was exponential, best described by an inverse log-log equation with no break point (P < 0.001), indicating that the exponential curve did not tend toward a low plateau. PTH was higher for a given 25OH-D and decreased less with increasing 25OH-D in Shenyang than in Cambridge (country-ln25OH-D interaction, P = 0.0005). After adjusting for bone area, weight, height, age, and sex, hip bone mineral content (BMC) was significantly related to PTH concentration in Cambridge but not in Shenyang [femoral neck coefficient: Cambridge = -0.064 (SE 0.027), P = 0.02; Shenyang = -0.027 (0.028), P = 0.3; trochanter: Cambridge = -0.116 (0.034), P = 0.001; Shenyang = -0.019 (0.027), P = 0.5]. There was a significant country lnPTH interaction at the trochanter (P = 0.02), but not at the femoral neck (P = 0.7). A weak positive association between BMC at the femoral neck and 25OH-D concentration was found in Cambridge [coefficient: 0.054 (0.028), P = 0.05] but not in Shenyang (coefficient: -0.013, P = 0.5; country-ln25OH-D interaction, P = 0.07). Urinary DPD concentration was also positively related to plasma PTH concentration in Cambridge subjects only [coefficient: 0.2 (0.08), P = 0.02]. These data suggest that although PTH increases when 25OH-D decreases, and Chinese people have a higher PTH for a given 25OH-D, older Chinese adults may be more resistant than Britons to the effects of PTH on bone. PMID- 14555268 TI - Homologous growth hormone accelerates bone healing--a biomechanical and histological study. AB - The purpose of this study was to prove whether homologous growth hormone has a beneficial effect in the early phase of bone healing. Therefore the left tibias of 24 Yucatan micropigs were osteotomized and stabilized by plate fixation. The treatment group (12 animals) received 100 microg of recombinant porcine growth hormone (rpGH)/kg body w/day sc, whereas the control pigs (12 animals) received 1 ml sodium chloride as placebo. After a healing period of 4 weeks the animals were sacrificed and destructive torsional testing was performed. For histological evaluation 6 microm serial slices of the tibiae were stained with von Kossa. The total area of callus formation (CA) and the mineralized bone area (BA) were quantified by image analysis. The fraction of mineralized bone tissue within the callus area, the bone density (BD), was calculated as follows: BD = (BA/CA) x 100. Torsional failure load was 91% higher and torsional stiffness 61% higher in the treatment group than in the control group (P < 0.05). The histomorphometric measurements revealed an advance for the CA (GH: 127.6 +/- 38.9 mm(2); placebo: 75.9 +/- 50.7 mm(2); P < 0.005) as well as for the BA (GH: 89.3 +/- 25.8 mm(2); placebo: 55.9 +/- 38.5 mm(2); P < 0.001) for the GH-treated animals in comparison to the control animals. The BD was similar in both groups (GH: 70.6 +/- 8.4%; placebo: 74.0 +/- 6.24%; P = 0.28). These data indicate that administration of homologous GH stimulates callus formation and ossification in the early phase of bone healing, which consequently results in an increased mechanical strength and stiffness. PMID- 14555269 TI - Growth hormone substitution increases gene expression of members of the IGF family in cortical bone from women with adult onset growth hormone deficiency- relationship with bone turn-over. AB - OBJECTIVE: To investigate the effects of growth hormone (GH) replacement therapy on bone matrix gene expression of insulin-like growth factors (IGFs) and markers of bone metabolism in women with adult-onset GH deficiency (GHD). DESIGN AND METHODS: Nineteen women, mean age 45 (range 24-56) years, were included in a double-blind, placebo-controlled parallel group study for 12 months. Biochemical markers were measured at baseline, 6 and 12 months. Bone biopsies were obtained and BMD was measured at baseline and after 12 months. RESULTS: Maximum responses were observed after 6 and 12 months, for bone resorptive and bone formative markers respectively. GH therapy enhanced gene expression in cortical bone of IGFs, GH-and calcitonin-receptor (CR) and osteoprotegerin (OPG), however with the most pronounced effects on CR and IGF-I. Changes in IGF-I gene expression during longitudinal follow-up were significantly correlated with changes in both circulating IGF-I (r = 0.82, p < 0.05), changes in markers of enhanced osteoclastic activity, measured both locally in bone (CR, r = 0.87, p < 0.01) and in serum (CTX-I, r = 0.86, p < 0.05), as well as serum bone ALP (r = 0.96, p < 0.01). CONCLUSIONS: This study indicates that both liver- and bone-derived IGF-I may be significant in mediating the effects of GH on bone metabolism in humans. PMID- 14555270 TI - Adiponectin as a novel determinant of bone mineral density and visceral fat. AB - Growing evidence suggests that positive associations between fat mass (FM) and bone mineral density (BMD) are mediated by not only biomechanical but also biochemical factors. Adiponectin is a novel adipocyte-derived hormone that regulates energy homeostasis and has anti-inflammatory and anti-atherogenic effects. Unlike other adipokines such as leptin, adiponectin levels decrease in obesity and type 2 diabetes. The purpose of our study was to investigate associations of serum adiponectin with BMD (DXA and QCT), FM (DXA and QCT), and serum leptin and soluble leptin receptor levels in 38 women and 42 men (age 39 81, BMI 17-55, 86% with type 2 diabetes). After adjusting for age, gender, race, smoking, and diabetes status, serum adiponectin was inversely associated with areal BMD (r = -0.20 to -0.3, all P < 0.01), volumetric BMD (r = -0.35 to -0.44, all P < 0.01), and visceral fat volume (r = -0.30, P < 0.01). These associations remained significant after adjusting for whole body fat mass. The associations of adiponectin with subcutaneous fat volume, whole body FM, and serum leptin level were not significant (all P > 0.1). These data suggest that adiponectin may play a role in the protective effects of visceral fat on BMD. PMID- 14555271 TI - Lovastatin inhibits adipogenic and stimulates osteogenic differentiation by suppressing PPARgamma2 and increasing Cbfa1/Runx2 expression in bone marrow mesenchymal cell cultures. AB - The mechanism whereby lovastatin can counteract steroid-induced osteonecrosis and osteoporosis is poorly understood. We assessed the effect of lovastatin on a multipotential cell line, D1, which is capable of differentiating into either the osteoblast or the adipocyte lineage. The expression of bone cell and fat cell transcription factors Cbfa1/Runx2 and PPARgamma2, respectively, were determined. 422aP2 gene expression was analyzed. Osteocalcin promoter activity was measured by cotransfecting the cells with the phOC-luc and pSV beta-Gal plasmids. Lovastatin enhanced osteoblast differentiation as assessed by a 1.8x increase in expression of Cbfa1/Runx2 and by a 5x increase in osteocalcin promoter activity. Expression of PPARgamma2 was decreased by 60%. By enhancing osteoblast gene expression and by inhibiting adipogenesis, lovastatin may shunt uncommitted osteoprogenitor cells in marrow from the adipocytic to the osteoblastic differentiation pathway. Future evaluation of lovastatin and other lipid-lowering drugs will help determine their potential as therapeutic agents for osteonecrosis and osteoporosis. PMID- 14555272 TI - Localized insulin-like growth factor I delivery to enhance new bone formation. AB - Insulin-like growth factor I (IGF I) exerts an important role during skeletal growth and bone formation. Therefore, its localized delivery appears attractive for the treatment of bone defects. To prolong IGF I delivery, we entrapped the protein into biodegradable poly(lactide-co-glycolide) microspheres (PLGA MS) and evaluated the potential of this delivery system for new bone formation in two defect models of ovine long bones, i.e., a 8-mm methaphyseal drill hole and a 10 mm segmental tibia defect. Administration of 100 microg of IGF I in PLGA MS resulted in new bone formation within 3 weeks in the drill hole and bridging of the segmental defect within 8 weeks. The observed increase of 12% newly formed bone in the drill hole defect after 3 weeks was substantial, compared to the measured morphometric bone-to-total area ratio of 31% bone in normal cancellous bone. Bone regeneration was further explored by measuring gene expression of typical markers for local mediators and growth factors by real-time polymerase chain reaction. Inflammation was reduced in presence of IGF I and this in vivo observation was corroborated in vitro by quantifying gene expression of inflammatory proteins and by assessing the activation of the NF-kappaB pathway, playing an important role in the regulation of inflammation. Administration of the IGF I delivery system downregulated inflammatory marker gene expression at the site of bone injury, induced new bone formation and reduced bone resorption, and resulted in bridging of 10-mm segmental tibial defects within 8 weeks. PMID- 14555273 TI - Endothelin-1 promotes osteoprogenitor proliferation and differentiation in fetal rat calvarial cell cultures. AB - Endothelin-1 (ET-1), a peptide produced by vascular endothelial cells, has been suggested to be one of the signaling factors between vascular and osteoblastic cells during bone growth and remodeling. The osteoinductive effects of ET-1 were tested on fetal rat calvaria which have the ability to form bone nodules in culture. ET-1 (10(-10) to 10(-6) M) dose-dependently increased cell proliferation. The effect of ET-1 (10(-8) M) on proliferation was greater than that of dexamethasone (Dex; 10(-8) M). ET-1 also increased the number of bone nodules by 146% over untreated cells, which coincided with a 3.1-fold increase in alkaline phosphatase activity. Limiting dilution assays showed that ET-1 treatment increased the number of osteoprogenitors (CFU-AP and CFU-OB) beyond what would be expected by a proliferative effect alone, indicating that ET-1 also stimulated osteoblast differentiation. Osteocalcin mRNA expression was upregulated as shown by Northern blot analysis. Using cDNA microarray analysis, ET-1 treatment resulted in an expression profile that included an upregulation of 163 genes and expressed sequence tags. Simultaneous addition of ET-1 and Dex to the medium further increased the number of bone nodules and alkaline phosphatase activity over either treatment alone. Our results show that ET-1 promotes both osteoblastic proliferation and differentiation and that the effects of ET-1 and Dex on differentiation are cooperative. PMID- 14555274 TI - Cyclic tensile strain and cyclic hydrostatic pressure differentially regulate expression of hypertrophic markers in primary chondrocytes. AB - Endochondral ossification is regulated by many factors, including mechanical stimuli, which can suppress or accelerate chondrocyte maturation. Mathematical models of endochondral ossification have suggested that tension (or shear stress) can accelerate the formation of endochondral bone, while hydrostatic stress preserves the cartilage phenotype. The goal of this study was to test this hypothesis by examining the expression of hypertrophic chondrocyte markers (transcription factor Cbfa1, MMP-13, type X collagen, VEGF, CTGF) and cartilage matrix proteins under cyclic tension and cyclic hydrostatic pressure. Chondrocyte seeded alginate constructs were exposed to one of the two loading modes for a period of 3 h per day for 3 days. Gene expression was analyzed using real-time RT PCR. Cyclic tension upregulated the expression of Cbfa1, MMP-13, CTGF, type X collagen and VEGF and downregulated the expression of TIMP-1. Cyclic tension also upregulated the expression of type 2 collagen, COMP and lubricin, but did not change the expression of SOX9 and aggrecan. Cyclic hydrostatic pressure downregulated the expression of MMP-13 and type I collagen and upregulated expression of TIMP-1 compared to the unloaded controls. Hydrostatic pressure may slow chondrocyte differentiation and have a chondroprotective, anti-angiogenic influence on cartilage tissue. Our results suggest that cyclic tension activates the Cbfa1/MMP-13 pathway and increases the expression of terminal differentiation hypertrophic markers. Mammalian chondrocytes appear to have evolved complex mechanoresponsive mechanisms, the effects of which can be observed in the histomorphologic establishment of the cartilaginous skeleton during development and maturation. PMID- 14555275 TI - Transcriptional induction of connective tissue growth factor/hypertrophic chondrocyte-specific 24 gene by dexamethasone in human chondrocytic cells. AB - Connective tissue growth factor (CTGF/Hcs24) is a critical growth factor for chondrocytic growth and differentiation. In this report, we describe for the first time glucocorticoid-mediated induction of the CTGF/Hcs24 gene in a chondrocytic cell line, HCS-2/8. Steady-state mRNA levels of CTGF/Hcs24 were remarkably increased after treatment with 50 nM dexamethasone, as confirmed by Northern blotting and quantitative real-time polymerase chain reaction (PCR) analysis. Corresponding to the increase in mRNA, production of CTGF/Hcs24 protein was remarkably enhanced, following a time course of up to 6 h. The observed increase in mRNA can be ascribed to transcriptional enhancement, since the stability of CTGF/Hcs24 mRNA was not affected by the same concentration of dexamethasone, which was indicated by the results of an mRNA degradation assay. However, unexpectedly, the prototypic ctgf/hcs24 promoter was not responsible for the dexamethasone stimulation, suggesting the glucocorticoid receptor binding site(s) to be elsewhere in the CTGF/Hcs24 gene. Enhancement of the prototypic promoter activity by dexamethasone was observed in murine fibroblastic cells, demonstrating the complexity of the regulatory mechanism of ctgf/hcs24 gene expression. Of importance, dexamethasone at the same concentration significantly stimulated proteoglycan synthesis in HCS-2/8 cells up to the same levels as exogenously added CTGF/Hcs24. These findings represent a novel effect of glucocorticoid on the production of CTGF/Hcs24 by chondrocytic cells, and indicate that CTGF/Hcs24 may mediate the stimulative effect of dexamethasone on chondrocytic phenotypes. Also, our results shed light on the complex mechanism of CTGF/Hcs24 induction by glucocorticoids. PMID- 14555276 TI - Hyaluronic acid reverses the abnormal synthetic activity of human osteoarthritic subchondral bone osteoblasts. AB - The underlying mechanisms responsible for both cartilage loss and subchondral bone changes in osteoarthritis (OA) remain unknown. It is becoming recognized that the extracellular matrix influences the metabolism of cells both in vivo and in vitro and can modify their responses to external stimuli. Indeed, the glycosaminoglycan/proteoglycan matrix is of major importance for the proliferation and/or differentiation of a number of cells. Here, we determined the potential role of hyaluronic acid (HA) of increasing molecular weight (MW) to alter the expression of metabolic markers and cytokine production by human osteoarthritic (OA) subchondral osteoblasts (Ob). Both 1,25(OH)(2)D(3)-induced alkaline phosphatase activity (ALPase) and osteocalcin release were increased in OA Ob when compared to normal. HA reduced osteocalcin release in OA Ob at MW of 300 and above, whereas HA failed to significantly modify ALPase. Parathyroid hormone (PTH) stimulated cyclic AMP (cAMP) formation by OA Ob. HA had a biphasic effect on this PTH-dependent activity, totally inhibiting cAMP formation at MW of 300 and 800. HA of increasing MW progressively reduced the levels of Prostaglandin E(2) (PGE(2)) and interleukin-6 (IL-6) produced by OA Ob. Interestingly, urokinase plasminogen activator (uPA) and and PA inhibitor-1 (PAI 1) levels were not significantly affected by HA of increasing MW; however, the PAI-1 to uPA ratio showed a slight, yet nonsignificant increase. Surprisingly, uPA activity was increased in OA Ob under the same conditions. Last, HA had no effect on the production of insulin-like growth factor-1 by these cells. Our data suggest that high MW HA can modify cellular parameters in OA Ob that are increased when compared to normal. The effect of HA on inflammatory mediators, such as PGE(2) and IL-6, and on uPA activity is more striking at higher MW as well. Taken together, these results could suggest that HA of increasing MW has positive effects on OA Ob by modifying their biological synthetic capacities. PMID- 14555277 TI - Resting T cells negatively regulate osteoclast generation from peripheral blood monocytes. AB - There is accumulating evidence that T cells may be involved in osteoclastogenesis in a variety of murine systems. However, the precise role of human T cells in the regulation of osteoclast generation is still unclear. To address this issue, we investigated the effect of resting peripheral T cells on receptor activator of NF kappaB ligand (RANKL)-induced osteoclast generation from human peripheral monocytes. Although osteoclasts were not generated in the culture of human peripheral blood mononuclear cells (PBMC) in the presence of RANKL and macrophage colony-stimulating factor (M-CSF), the addition of cyclosporine A (CsA), a potent inhibitor of T-cell function, resulted in the formation of an increasing number of lacunae resorption on dentine, suggesting T cells may inhibit osteoclast formation. In a coculture of T cells and monocytes, which were isolated from PBMC, T cells inhibited the osteoclast generation from monocytes, as determined by tartrate-resistant acid phosphatase (TRAP) staining and a pit assay using dentine. This inhibition of osteoclast generation by T cells was also observed in a culture of the parathyroid hormone-stimulated SaOS4/3 osteoblast cell line and monocytes. The culture in Transwell plates revealed that the cell-to-cell interaction was not required for the inhibition, suggesting that T-cell cytokines may be responsible for the inhibition. Among inhibitory T-cell cytokines on osteoclastogenesis, granulocyte-macrophage colony-stimulating factor (GM-CSF) and interferon-gamma (IFN-gamma) were actively produced by CD4 T cells but not CD8 T cells in the coculture of T cells with monocytes, and the neutralizing antibodies to these cytokines partially rescued the T-cell-induced inhibition of osteoclast formation. Although CsA did not affect RANKL-induced osteoclast generation in the culture of monocytes alone, it completely rescued the T-cell-induced inhibition of osteoclast formation and strongly inhibited the production of GM-CSF and IFN gamma. Thus, we demonstrate that resting T cells negatively regulate the osteoclast generation via production of GM-CSF and IFN-gamma by CD4 T cells and that CsA stimulates the osteoclast generation through the inhibition of the production of these cytokines. These findings provide new insight into therapeutic strategies for immunosuppression-induced bone loss in transplant and other diseases. PMID- 14555278 TI - Inhibition of RANKL-induced osteoclast formation in mouse bone marrow cells by IL 12: involvement of IFN-gamma possibly induced from non-T cell population. AB - IL-12 was shown to have the potential to inhibit osteoclast formation in mouse bone marrow cells treated with macrophage colony-stimulating factor (M-CSF) and receptor activator of NF-kappaB ligand (RANKL). When bone marrow macrophages (BMM) were used as osteoclast precursors, IL-12 failed to inhibit M-CSF/RANKL induced osteoclast formation from BMM. In coculture experiments using transwells, IL-12 did inhibit osteoclast formation from BMM cocultured with whole bone marrow cells. These results indicated that IL-12 indirectly affected M-CSF/RANKL-induced osteoclastogenesis in bone marrow cells and that the inhibition of IL-12 on osteoclast formation was caused by a humoral factor from bone marrow cells treated with IL-12. Experiments with anti-interferon (IFN)-gamma antibody and bone marrow cells from IFN-gamma receptor knockout mice revealed that IFN-gamma might be involved in the inhibition of osteoclast formation in this system. The expression of osteoprotegerin mRNA in bone marrow cells was not affected by treatment with IL-12. The inhibitory effect of IL-12 on osteoclast formation was also seen in the T cell-depleted bone marrow cells of normal mice and the whole bone marrow cells of athymic nude mice, while the inhibitory effect of IL-12 was partially suppressed in the B cell-depleted bone marrow cells. The inhibitory effect of IL-12 on M-CSF/RANKL-induced osteoclastogenesis was not accompanied with cell death, in contrast with our previous finding that the inhibitory effect of IL-12 on M-CSF/TNF-alpha-induced osteoclastogenesis is attributable to Fas and FasL-mediated apoptosis. PMID- 14555279 TI - Inhibition of osteoblast apoptosis by thrombin. AB - The multifunctional serine protease thrombin has been shown to be a specific agonist for a variety of functional responses of cells including osteoblasts. The current study was conducted to determine if thrombin was capable of inhibiting apoptosis in osteoblasts, and if so, to examine the mechanism by which this occurred. Thrombin (20-100 nM) significantly inhibited apoptosis in serum-starved cultures of the human osteoblast-like Saos-2 cell line and cultures of primary osteoblasts isolated from mouse calvariae, as well as dexamethasone-treated primary mouse osteoblasts. Inhibition of serum deprivation-induced apoptosis was shown to require thrombin's specific proteolytic activity. Primary mouse osteoblasts were found to express two functional thrombin receptors, PAR-1 and PAR-4. Thrombin inhibited serum deprivation-induced apoptosis in osteoblasts isolated from PAR-1 null mice to the same degree as in osteoblasts isolated from wild-type mice. Treatment of serum-deprived osteoblasts, isolated from either PAR 1 null or wild-type mice, with a PAR-4-activating peptide failed to significantly inhibit apoptosis compared to the relevant control. Medium conditioned by thrombin-treated osteoblasts, in which thrombin had been inactivated, was able to inhibit serum deprivation-induced osteoblast apoptosis almost as well as thrombin itself. Blocking protein synthesis, by cycloheximide pretreatment of the conditioning cells, prevented this action. The ability of known osteoblast survival factors, such as transforming growth factor beta1, fibroblast growth factor-2, insulin-like growth factor-II, and interleukin-6, to inhibit serum deprivation-induced osteoblast apoptosis was also tested. None of these factors was able to inhibit serum deprivation-induced osteoblast apoptosis to the same extent as thrombin. The results presented here demonstrate that thrombin treatment of osteoblasts inhibits apoptosis induced either by dexamethasone or by serum deprivation. Furthermore, it does so independently of the known thrombin receptors by bringing about the synthesis and/or secretion of an unknown survival factor or factors, which then act in an autocrine fashion to inhibit apoptosis. PMID- 14555280 TI - Finite element models predict in vitro vertebral body compressive strength better than quantitative computed tomography. AB - The correlation between bone mineral density and vertebral strength is not based on mechanical principles and thus the method cannot reflect the effects of subtle geometric features and densitometric inhomogeneities that may substantially affect vertebral strength. Finite element models derived from quantitative computed tomography (QCT) scans overcome such limitations. The overall goal of this study was to establish that QCT-based "voxel" finite element models are better predictors of vertebral compressive strength than QCT measures of bone mineral density with or without measures of cross-sectional area. QCT scans were taken of 13 vertebral bodies excised from 13 cadavers (L1-L4; age: 37-87 years; M = 6, F = 7) and used to calculate bone mineral density (BMD(QCT)). The QCT voxel data were converted into linearly elastic finite element models of each vertebra, from which measures of vertebral stiffness and strength were computed. The vertebrae were biomechanically tested in compression to measure strength. Vertebral strength was positively correlated with the finite element measures of strength (r(2) = 0.86, P < 0.0001) and stiffness (r(2) = 0.82, P < 0.0001), the product of BMD(QCT) and vertebral minimum cross-sectional area (r(2) = 0.65, P = 0.0008), and BMD(QCT) alone (r(2) = 0.53, P = 0.005). These results demonstrate that highly automated "voxel" finite element models are superior to correlation based QCT methods in predicting vertebral compressive strength and therefore offer great promise for improvement of clinical fracture risk assessment. PMID- 14555281 TI - Histamine H3 receptors regulate vascular permeability changes in the skin of mast cell-deficient mice. AB - The participation of histamine H(3) receptors in the regulation of skin vascular permeability changes in mast cell-deficient mice was studied. Although intradermal injection of histamine H(3) antagonists, iodophenpropit and clobenpropit, at a dose of 100 nmol/site caused significant increases in skin vascular permeability in both mast cell-deficient (WBB6F1 W/W(v)) and wild-type (WBB6F1 +/+) mice, this response was significantly lower in mast cell-deficient mice than in the wild-type controls. Histamine also caused dose-related increases in skin vascular permeability in both wild-type and mast cell-deficient mice. Significant effects were observed at doses of 10 and 100 nmol/site, and no significant difference in skin vascular permeability was observed between mast cell-deficient and wild-type mice. However, histamine contents of dorsal skin in mast cell-deficient mice were significantly lower than in wild-type mice. In addition, the H(1) antagonists diphenhydramine and chlorpheniramine and the NK(1) antagonists, L-732,138 and L-733,060, were able to antagonize H(3) antagonist induced skin vascular permeability. These results indicated that blockade of H(3) receptors by H(3) antagonists induce skin vascular permeability through mast cell dependent mechanisms. In addition, histamine and, to a lesser extent substance P are involved in the reaction. PMID- 14555282 TI - The [173-196] fragment of ovalbumin suppresses ovalbumin-specific rat IgE responses. AB - Peptides and protein hydrolysates are attractive tools for the induction of tolerance or regulation of targeted B and/or T cell responses. In vivo, peptides are mainly produced by the action of digestive enzymes or following the processing of exogenous antigens by antigen-presenting cells (APCs). In vitro, these molecules are generally produced by enzymatic digestion and chemical hydrolysis of proteins. We investigated the T and B cell determinants of the major food allergen ovalbumin (nOVA) in rat by analyzing (1) the stimulatory effect of nOVA peptides generated by cyanogen bromide (CNBr) cleavage on nOVA specific T cells, and (2) the potential of CNBr-derived OVA fractions to induce oral tolerance to nOVA. Peptide fractions of the CNBr-hydrolysated OVA were isolated by high-pressure liquid chromatography and tested for their ability to stimulate nOVA-specific T cells isolated from rats parenterally immunized with nOVA. The nOVA fractions containing the stimulatory determinants were then intragastrically administered to rat to test their potential to induce oral tolerance. The hole CNBr hydolysate stimulated proliferation of nOVA-specific T cells. Three out of the five HPLC-purified peptidic fractions were also able to stimulate proliferation and cytokine production by nOVA-specific T cells. A peptide fraction exhibiting a single peak by HPLC contained the 173-196 nOVA segment and stimulated nOVA-specific T cells. This segment also promoted oral tolerance to nOVA and reduced IgE responses. CNBr hydrolysis releases several peptides with stimulatory effect on nOVA-specific T cells including a new nOVA [173-196] T cell determinant which induces oral tolerance to nOVA. PMID- 14555283 TI - Nicorandil inhibits the release of TNFalpha from a lymphocyte cell line and peripheral blood lymphocytes. AB - Modulation of cytokine release may be of interest in modulating inflammatory diseases. This study determined whether nicorandil, a potassium channel opener, and nitric oxide (NO) donor could inhibit the release of tumour necrosis factor alpha (TNFalpha) from lymphocytes. Nicorandil significantly and dose-dependently inhibited the TNFalpha release from a human Epstein Barr virus-transformed B lymphocyte cell line (EBV-B) and peripheral blood B and T lymphocytes. The inhibition was reversed by the addition of both potassium channel inhibitor glibenclamide and the guanylyl cyclase inhibitor 1H-(1,2,4) oxadiazolo (4,3) quinoxalin-1-one (ODQ). Other potassium channel openers, pinacidil, or the nicorandil analogue SG-209, however, failed to demonstrate inhibition of TNFalpha release. The NO scavenger haemoglobin was unable to reverse the nicorandil induced TNFalpha inhibition, but in contrast to this, sodium nitroprusside (SNP) partially inhibited the release, which was reversed by haemoglobin. Nicorandil is able to inhibit TNFalpha release from lymphocytes, which requires the dual modes of both potassium channel opening and the nitrate moiety. Moreover, NO donation mechanism appears to be more dominant in the nicorandil inhibitory activity in lymphocytes.The dual mechanism involved in the inhibition of this cytokines may represent a novel therapeutical approach in the modulation of inflammatory disease. PMID- 14555285 TI - Bisphenol A significantly enhances the neutrophilic differentiation of promyelocytic HL-60 cells. AB - Bisphenol A (BPA) is a well-known endocrine disruptor. However, little information is available on its immunological effects. To investigate the effect of BPA on leukocyte differentiation, we investigated its action on the neutrophilic differentiation of HL-60 cells induced by dimethylsulfoxide and granulocyte colony-stimulating factor (G-CSF) for 6 days. At low concentrations (10(-10)-10(-8) M), BPA significantly increased the superoxide production by differentiated HL-60 cells stimulated with opsonized zymosan (OZ) by about 20%, and expression of CD18, a component of the OZ-receptor, was increased to a similar extent by 10(-9) M BPA. To investigate the effect of BPA on the activity of PU.1, a transcription factor specific for granulocytic differentiation, we established a stable clone that expressed luciferase as a reporter of PU.1 activity. PU.1 activity increased during the neutrophilic differentiation of HL 60 cells, reaching a peak on day 3 and decreasing thereafter. Nanomolar BPA augmented the PU.1 activity on day 3 by about 60%. On the other hand, tamoxifen, a competitive inhibitor of estrogen receptors, did not suppress the effect of BPA on the differentiation of HL-60 cells. These results suggest that BPA exerts an enhancing effect on the neutrophilic maturation of leukocytes through an estrogen receptor-independent pathway. Long-term exposure to BPA might significantly affect the innate immunity of mammals, even at low doses. PMID- 14555284 TI - Reversal of T cell anergy in leprosy patients: in vitro presentation with Mycobacterium leprae antigens using murabutide and Trat peptide in liposomal delivery. AB - Mycobacterium leprae, the causative agent of leprosy resides and multiplies within the host monocytes and macrophages, thereby evading host immune system. Cell-mediated immune response (CMI) plays a vital role as evidenced from the high CMI in BT/TT (borderline and tuberculoid) patients and conversely low in BL/LL (borderline and lepromatous) patients. In the present study, an attempt was made to immunomodulate the anergized T cells of lepromatous leprosy patients by presenting the mycobacterial antigen in combination with T cell adjuvant, murabutide (active analog of muramyl' dipeptide, MDP-BE) and a Trat peptide (T cell epitope of Integral membrane protein (Trat) from Escherichia coli) in particulate form (liposomes) or soluble form (media). PBMNC of normal, BT/TT and BL/LL were stimulated in vitro with five mycobacterial antigens (Ag) in the following formulations, Ag, Ag+murabutide, Ag+murabutide+Trat peptide either in liposomes or in medium. All the five antigen(s) when delivered in liposomes containing murabutide and Trat peptide showed a very high lymphoproliferative response (p<0.001) in all the three groups. IFN-gamma and IL-2 were significantly (p<0.001) high in these culture supernatants compared to IL-10 and IL-4 confirming a shift from CD4+Th2 to Th1 response in leprosy patients with particulate mode of antigen presentation. Interestingly, PBMNC derived from lepromatous patients also showed consistent T cell proliferation with all the formulations. Further, the mechanism of liposomal processing of antigens was studied using different inhibitors that interfere at different stages of antigen presentation. Results indicate that this study may pave way for an immunotherapeutic approach for reverting the anergic T cells of lepromatous patients to proliferating T cells with the release of Th1 cytokines thereby restoring the CMI response in these patients. PMID- 14555286 TI - Cytokine release by murine spleen cells following multiple low dose streptozotocin-induced diabetes and treatment with a TNFalpha transcriptional inhibitor. AB - We recently reported that administration of 9-[(1R, 3R)-trans-cyclopentan-3-ol] adenine (MDL 201,449A), a transcriptional inhibitor of TNFalpha, decreased hyperglycemia in murine diabetes induced by multiple low doses of streptozotocin (MLDSTZ). In the present study, we first investigated if in vivo administration of MDL 201,449A in the MLDSTZ model affects cytokine release from cultured spleen cells. Secondly, we studied how MDL 201,449A affects cytokine release from normal cultured spleen cells. In all experiments, the mitogen concanavalin A (2 micro g/ml) was added to the cultured spleen cells in order to enhance cytokine release. MLDSTZ treatment in vivo caused increased IFNgamma secretion, a decreased/retarded rate of increased TNFalpha accumulation, whereas IL-10 production was not altered compared to vehicle-treated mice. MDL 201,449A treatment of MLDSTZ mice did not affect cytokine release from spleen cells subsequently cultured in the absence of MDL 201,449A. We also studied cytokine release from normal spleen cells in the presence or absence of MDL 201,449A. Production of TNFalpha, IFNgamma and IL-10 was all suppressed by the drug. In groups where exposure to MDL 201,449A was discontinued, cytokine levels increased promptly and in the case of TNFalpha secretion, it exceeded the production from control cells. Our data suggest an enhanced Th1 cytokine secretion from spleen cells derived from MLDSTZ-treated mice. MDL 210,449A may be a potent inhibitor of cytokine secretion, albeit not completely selective for TNFalpha. However, when MDL 201,449A is withdrawn, there may be a rebound phenomenon of increased TNFalpha secretion. PMID- 14555287 TI - Comparison of effects of Y-27632 and Isoproterenol on release of cytokines from human peripheral T cells. AB - Y-27632 selectively inhibits Rho-associated protein kinases (ROCKs), an effector for RhoA. The RhoA system is involved in T cell activation. Y-27632 mimicked effects of beta agonists on human cells. We examined the effects of both Y-27632 and Isoproterenol (Iso) on the release of T helper type 1 (Th-1) cytokines (interleukin (IL)-2 and interferon (IFN)-gamma) and Th-2 cytokines (IL-4 and IL 5) from activated human T cells. Peripheral T cells obtained from seven healthy volunteers were incubated in the presence of Y-27632 (0.1-10 micro M) for 30 min, and stimulated with 50 micro g/ml of Concanavalin A (Con A) for 24 h. Concomitantly, after an incubation with medium alone, cells were stimulated with Con A in the presence of Iso (0.1-10 micro M). The concentration of these cytokines in supernatants was measured by ELISA. Both Y-27632 and Iso suppressed release of Th-1 cytokines, decreased release of Th-2 cytokines weakly, and reduced ratio of Th-1/Th-2 cytokine release from Con A-activated T cells. These inhibitory effects of Y-27632 closely resembled those of Iso at each concentration tested. Y-27632 mimicked effects of Iso on secretion of Th-1 and Th 2 cytokines from human peripheral T cells activated with Con A. It is suggested that the RhoA/ROCK system plays an important role in the release of Th-1 cytokines and is partially involved in the release of Th-2 cytokines from human T cells activated through T cell receptor (TCR). PMID- 14555288 TI - Nitric oxide synthase inhibitors modulate lipopolysaccharide-induced hepatocyte injury: dissociation between in vivo and in vitro effects. AB - Effects of endotoxemia-induced NO production on rat liver and hepatocytes in culture were investigated. Rats were treated intraperitoneally with saline, lipopolysaccharide (LPS, 10 mg/kg), L-nitroarginine methyl ester (L-NAME)+LPS, aminoguanidine (AG)+LPS, FK 506+LPS, S-nitroso-N-acetyl penicillamine (SNAP)+L NAME+LPS and SNAP+FK 506+LPS. Mortality, hepatocyte viability and liver function test were estimated. Liver morphology was observed by light and electron microscopy. Hepatocyte cultures were treated with LPS, cytokine mixture (CM) with or without FK 506, L-NAME or AG. Hepatocyte function and inducible form of NOS (iNOS) expression were evaluated. Twenty-four hours after treatments with saline, LPS, L-NAME+LPS, AG+LPS, FK 506+LPS, SNAP+L-NAME+LPS and SNAP+FK 506+LPS, rat mortalities were 0%, 10%, 48%, 8%, 20%, 38% and 0%, and hepatocyte viabilities were 93+/-3%, 80+/-3%, 52+/-8%, 88+/-1%, 70+/-3%, 80+/-4% and 82+/-3%, respectively. AG+LPS or L-NAME+LPS administration was followed by excessive vacuolization of hepatocytes with lesions in the intermediary lobule zone characterized by features of secondary necrosis as a continuation of apoptotic processes. SNAP+L-NAME+LPS resulted in a well-preserved structure of central vein lobules with sparse signs of apoptosis. Treatment with LPS or CM increased iNOS expression in hepatocyte culture, which was inhibited by L-NAME, FK 506 or AG. AG reduced LPS-induced rise in alanine aminotransferase leakage. LPS-induced NO exerts cytoprotective effects in vivo, while LPS-induced NO in vitro appears to be toxic. Based on the data of this report, one cannot use in vitro results to predict in vivo responses to LPS-induced NO production. The pharmacological modulation of iNOS expression or NO production in vivo or in vitro, therefore, by the development of specific NO donors or inhibitors is promising for improvement of hepatocyte functions under the two experimental conditions, respectively. PMID- 14555289 TI - Reduced birth defects caused by maternal immune stimulation may involve increased expression of growth promoting genes and cytokine GM-CSF in the spleen of diabetic ICR mice. AB - Maternal immune stimulation in mice decreases fetal abnormalities caused by diverse etiologies. Growth factors produced by activated immune cells were proposed to be key mediators that may exert their effects on placenta or embryo. Diabetes disrupts the secretion of cytokines, which may associate with diabetic embryopathy. Three different methods of maternal immune stimulation that result in approximately equal reduction of diabetic embryopathy were used in the present studies: footpad injection with complete Freund's adjuvant (CFA), intraperitoneal (i.p.) injection with granulocyte-macrophage colony-stimulating factor (GM-CSF), or i.p. injection with interferon-gamma (IFN-gamma). A gene microarray was then used to examine expression of a selected gene panel in splenic leukocytes. We hypothesized that maternal immune stimulation may act by overcoming altered gene expression patterns of immune cells in the diabetic mice, which partially mitigates the teratogenic effect of diabetes. It further seemed likely that a shared profile of splenic gene expression changes induced by the different immune stimulation procedures may be identified and related to reduced teratogenesis. The three procedures produced a common altered gene expression profile. Significantly affected genes included apoptotic and anti-apoptotic genes, and genes controlling cellular proliferation, and likely reflect a state of immune activation. The GM-CSF gene was up-regulated by all three immune stimulation procedures. The protein product of this gene regulates placental development, and was recently associated with reduced cleft palate in immune-stimulated pregnant mice after exposure to urethane. These data suggest that further studies of GM CSF as mediator of reduced birth defects in teratogen-challenged, immune stimulated mice are warranted. PMID- 14555290 TI - Batimastat reduces Mycobacterium tuberculosis-induced apoptosis in macrophages. AB - In this study, we report evidences that Mycobacterium tuberculosis (MTB)-induced apoptosis in macrophages is reduced by a broad-spectrum hydroxamic acid-based matrix metalloproteinase (MMP) inhibitor, Batimastat (BB-94). In particular, we show that BB-94 administration to MTB-infected macrophages inhibits apoptosis and the downmodulation of membrane CD14 expression. Moreover, the addition of broad spectrum matrix metalloproteinase inhibitor to cell culture, during MTB infection, decreases the release of soluble TNF-alpha and leads to a simultaneous increase of membrane TNF-alpha. These results show that MTB-induced apoptosis in macrophages is reduced by a MMP inhibitor and most probably is related to TNF alpha release. This identifies BB-94 as a simultaneous anti-apoptotic and anti inflammatory molecule during MTB infection. PMID- 14555291 TI - In vivo characterization of bioconjugate B cell toleragens with specificity for autoantibodies in antiphospholipid syndrome. AB - This study investigated the use of well-defined bioconjugate molecules to suppress antigen-specific B cell responses to domain I (DI) of human beta(2) glycoprotein I (beta(2)GPI) in rats. DI is the dominant target of pathogenic autoimmune antibodies in patients with antiphospholipid syndrome (APS), a disease characterized by antibody-mediated thromboembolic events. Rats primed with DI conjugated to keyhole limpet hemocyanin (DI-KLH) were rendered tolerant to subsequent antigen challenge by treatment with multivalent conjugates of DI. Antibodies to DI were suppressed 89-96% with intravenous doses of 500 micro g, and reductions were paralleled by decreases in splenic antigen-specific antibody forming cells (AFC). Suppression was achieved with a variety of conjugates having two to four copies of DI and circulating half-lives of 2.6-8.7 h. Antibodies to KLH were not suppressed, indicating the specificity of the approach. These results establish the basis for further development of therapeutic B cell toleragens to suppress pathogenic antibodies in APS and other autoimmune diseases. PMID- 14555292 TI - Pharmacological analysis of signal transduction pathways required for oxidative burst in chicken heterophils stimulated by a Toll-like receptor 2 agonist. AB - Toll-like receptors (TLRs) play an important role in the innate immune response of avian heterophils. We previously used the pharmacological inhibitors genistein, verapamil, chelerythrine, and pertussis toxin to investigate the upstream signaling events involved in TLR2-mediated oxidative burst in chicken heterophils. Only chelerythrine, a protein kinase C inhibitor, was found to significantly inhibit oxidative burst stimulated by the TLR2 agonist lipoteichoic acid (LTA). In the present study, we used selective pharmacological inhibitors to investigate the roles of phosphatidylinositol-3'-kinase (PI3-K), phospholipase C (PLC), calcium-dependent protein kinase C (PKC), extra-cellular signal regulated kinase (ERK), and nuclear translocation factor kappa B (NF-kappaB) on TLR2 mediated oxidative burst. U-73122 (a PLC inhibitor), wortmannin (a PI3-K inhibitor), PD 98059 (an ERK inhibitor), Go 6976 (a PKC inhibitor) and Bay 11 7082 (a NF-kappaB inhibitor) significantly decreased LTA-stimulated oxidative burst in heterophils by 77%, 30%, 36%, 78%, and 61%, respectively. Activated TLR2 utilizes PI3-K, PLC, PKC, ERK, and NF-kappaB as signaling factors that mediate the oxidative burst of chicken heterophils. PMID- 14555293 TI - Inhibition of Drynariae Rhizoma extracts on bone resorption mediated by processing of cathepsin K in cultured mouse osteoclasts. AB - In the traditional Korean medicine, Drynariae Rhizoma (DR) [Drynaria fortunei (kunze) J. Sm] has been reported as a good enhancer for bone healing. In this experiment, we investigate the effects of DR on bone resorption using the bone cells culture. Different concentrations of crude extract of DR were added to mouse bone cells culture. The mitochondria activity of the bone cells after exposure was determined by colorimetric MTT assay. It was demonstrated that DR has potential effects on the bone cells culture without any cytotoxicity. The most effective concentration of DR on bone cells was 100 micro g/ml. On the other hand, cathepsin K (Cat K) is the major cysteine protease expressed in osteoclasts and is thought to play a key role in matrix degradation during bone resorption. In this study, Mouse long bone cells including osteoclasts and osteoblast were treated with the PI3-kinase inhibitor, wortmannin (WT), and a specific inhibitor of protein kinase C (PKC), calphostin C. Although WT prevented the osteoclast mediated intracellular processing of Cat K, calphostin C did not. Similarly, treatment of osteoclasts-containing long bone cells with Drynariae Rhizoma (DR) extracts prevented the intracellular maturation of Cat K, suggesting that DR may disrupt the intracellular trafficking of pro Cat K. This is similar to that of WT. Since secreted proenzymes have the potential to reenter the cell via mannose 6-phosphate (M6P) receptor, to prevent this possibility, we tested WT and DR in the absence or presence of M6P. Inhibition of Cat K processing by WT or DR was observed in a dose-dependent manner. Furthermore, the addition of M6P resulted in enhanced potency of WT and DR. DR dose-dependently inhibited in vitro bone resorption with a potency similar to that observed for inhibition of Cat K processing. PMID- 14555294 TI - Assessment of peripheral arterial occlusive disease: comparison of multislice-CT angiography (MS-CTA) and intraarterial digital subtraction angiography (IA-DSA). AB - PURPOSE: The aim of the study was to assess the arterial vascular system of the lower extremities in patients with peripheral arterial occlusive disease using Multislice-CT angiography (MS-CTA) and to compare the results with the standard of reference, intraarterial digital subtraction angiography (IA-DSA). MATERIALS AND METHODS: MS-CTA and IA-DSA of the lower extremities were carried out on 23 patients with peripheral arterial occlusive disease (Fontain Stage IIb: 18, III: 3, IV: 2). MS-CTA comprised a 4 x 2.5 mm collimation, 15 mm table feed/rotation, 0.5 sec rotation time and 3 mm slice thickness (1.5 mm reconstruction increment). Delay time was determined by bolus tracking. 150 ml of contrast media were injected intravenously at a flow rate of 3 ml/sec. Maximum intensity projection (MIP) reformations were performed using a semi-automatic vessel tracking program. MS-CTA (axial and MIP-reformatted images) and IA-DSA were reviewed by two radiologists. The grade of vascular stenosis as well as occlusion were rated on a scale of 0 to 3 (0=0-50% stenosis, 1 = 51-75% stenosis, 2 = 76-99% stenosis, 3 = occlusion). RESULTS: For MS-CTA, the mean delay time was 30.2 s (23-40 s), mean scan time was 37.4 (33-42 s). Data analysis was based upon a total of 1136 vascular segments for both methods (568 each). A comparison of all the evaluated segments in both techniques revealed a MS-CTA / IA-DSA 86.3% match. Out of 442 segments proximal the trifurcation, 386 were correctly assessed in MS-CTA (87.3%) and distal the trifurcation, 101 out of 126 segments were rated correctly (80.2%). In MS-CTA, an overall confidence interval of 95% can be achieved in 83.2% to 89.0% for correctly rated stenosis grading. CONCLUSIONS: In patients with peripheral arterial occlusive disease, MS-CTA of the lower extremity is a promising minimal-invasive method for detection of relevant arterial stenoses. However, the technique was limited to routine diagnostic purposes due to severe calcifications and time consuming reconstruction procedures. PMID- 14555295 TI - Lysophosphatidic acid is the unique platelet-activating substance in human malignant ascites. AB - Pathological blood platelet activation promotes thrombosis in cancer patients, but the specific substances involved are still under investigation. Tumor exudates have been described to contain lysophosphatidic acid (LPA), a known platelet-activating substance, and the concentration of mediators present in malignant ascites are constantly equilibrated with the concentration in plasma. We hypothesized that the ascites of cancer patients might activate platelets, and that this may be caused by LPA. Indeed, ascites samples from 15 different patients with cancer induced shape change and an increase of cytosolic Ca2+ of isolated platelets; both responses were cross-desensitized by lysophosphatidic acid (LPA), but not by other platelet stimuli. Moreover shape change, Ca2+ mobilization and aggregation induced by ascites could be completely blocked by pretreatment of platelets with specific LPA-receptor antagonists. Phospholipids were extracted from ascites, separated by thin layer chromatography, and individual fractions were tested for activity on platelets. The platelet activating substance co-migrated with LPA, whereas other fractions were inactive. Notably, ascites induced through LPA-receptor activation platelet aggregation in whole blood. Our results suggest that LPA plays an essential role in the pathological platelet activation in cancer patients. We propose that LPA receptor antagonists could be effective in blocking cancer-associated platelet activation and thus preventing thrombosis. PMID- 14555296 TI - Evaluation of a didanosin-containing regimen including genotypic resistance testing: an open-label, multicenter study. AB - PURPOSE: To show Didanosin in a new formulation as a once-a-day capsula as a well tolerated and effective HIV-therapy when used in a protease sparing regimen including genotypic resistance pattern in blood, semen and cerebrospinal fluid before and during treatment. METHOD: Two groups of 58 patients, each containing 9 patients who had not been previously treated with any antiretroviral medication, and 49 patients heavily pretreated for 3,7 (DDI group) and 2,8 (non-DDI group) years, have been followed up for at least half a year. A group of 24 patients taking a special combination of Didanosin plus Efavirenz and Stavudine have been analysed with genotypic resistance testing concerning viral load response and resistance pattern under therapy. RESULTS: Suppression of plasma HIV-1 RNA to <50 copies/mL and <500 copies/mL in the DDI group was achieved in 74% and 84% of the pretreated patients, respectively, and in 100% of the naive patients after 24 weeks. In the non-DDI group suppression was achieved in 59% and 69% of the pretreated patients, respectively, and in also 100% of the naive patients. The viral load reduction in the DDI containing regimen at week 24 was 1.7 log subset 10 for the pretreated and 3,4 log subset 10 for the naive patients. In the non DDI group, the reduction was 1.5 for the pretreated and 4,0 for the naive patients. CD4 cell counts increased from 440 to 517 cells/microL at week 24 for the pretreated, and from 171 to 289 for the naive patients in the DDI containing regimen. In the other group, cells increased from 396 to 406 for the pretreated and from 155 to 321 for the naive patients. In each group, 12 patients discontinued treatment; 4 patients in the DDI group and 7 patients in the non-DDI group because of adverse events. There were no AIDS-defining events in the antiretroviral-treated patients in both groups. 16 patients of the special combination group (DDI, D4T and EFV) were evaluated for more than 24 weeks. Suppression of HIV-1 RNA to <50 copies /mL were found in 75% of the naive and 43% of the pretreated patients. No relevant mutations were found during treatment. CONCLUSION: The new formulation of Didanosin as a once-a-day capsula in a protease sparing regimen was well-tolerated, effective in reducing viral load and in preventing AIDS-defining events. The combination of DDI, D4T and EFV proved to be a potent therapy without developing relevant mutations. PMID- 14555297 TI - Lunar phase does not influence surgical quality. AB - INTRODUCTION: 10.5% of the German population believes in the effects of lunar phase on disease. The topic is hot in German TV program. It is believed that at new moon the rate of bleeding complications is increased and operations during the waning phase of the moon would be best to avoid complications, pain and scaring. To our knowledge the effect of lunar phase has not been studied in ambulatory surgery. PATIENTS AND METHODS: 782 patients were evaluated for complications and perception of the personal health after herniotomy, haemorrhoidectomy and crossectomy with partial vein stripping with or without phlebectomy as part of a quality control study. A questionnaire has been sent out to the patients asking the patient to rate postoperative pain, pain medication, restriction of daily activity, mental health and emotion, status of complaints after the operation. RESULTS: In 782 patients (mean age 50 years) 866 operations were performed. There were no major complications and only in 3.71% minor complications (local bleeding, haematoma, inflammation, abscess, seroma, lymphatic fistula, dehiscence) were observed. The operations were equally distributed to the lunar phases. Complications and patient's subjective perception of pain, restriction of daily activity, mental health and emotion, status of complaints after the operation were not associated with a lunar phase. CONCLUSIONS: The hypothesis that lunar phase influences the outcome of ambulatory operations is not valid. With regard to the organization of operations in the hospital and the patient's uncertainty to decide the right time the lunar phase philosophy may have an socio-economic impact not yet understood. PMID- 14555298 TI - Cognitive function and quality of diabetes care in patients with Type-2-diabetes mellitus in general practitioner practice. AB - INTRODUCTION/AIMS: The goal of the trial was the assessment of the quality of diabetes control and the cognitive function of all patients with type 2 diabetes mellitus treated in a randomly selected general practitioner practice in Thuringia, Germany. Furthermore possible interactions between patients' quality of diabetes control and their cognitive function should be analysed. PATIENTS AND METHODS: The investigation comprised all 141 patients with type 2 diabetes mellitus (age 67.3 +/- 10.5 years, body-mass index 29.3 +/- 4.5 kg/m2, patients without insulin therapy: n = 102, with insulin therapy: n = 39) who were patients at the general practitioner practice in Wandersleben, Thuringia, Germany, between November 1999 and April 2000. RESULTS: The mean HbA1c of all patients examined was 6.33 +/- 1.1% (normal range 3.5-6.8%). 55 patients were treated with diet (HbA1c 5.7 +/- 0.76%) and 47 patients were given oral antidiabetic drugs (HbA1c 6.5 +/- 0.76 %). 27/47 (57.4%) patients were treated with sulfonylurea, 37/47 (78,7%) with metformin, 3/47 (6,4%) with acarbose and 7/47 (14.9%) patients received glinides. 18/47 (38,3%) patients had combinations out of two or more oral antidiabetic drugs. A total of 39 patients with type 2 diabetes mellitus were treated with insulin (HbA1c 7.0 +/- 1.24%). The premorbide cognitive function of the patients was 97.9 +/- 10.0 IQ-points. It was significantly better than the actual cognitive function (91.8+/-13.5 IQ-points, p<0.001). The actual cognitive function showed a correlation with patient's age (r = -0.28, p = 0.001), educational level (r = -0.25, p = 0.006) and profession (r = -0.29, p = 0.001). Performing multivariate analysis the premorbide cognitive function (b = 0.53, p<0.001) and patient's age (b = -0.32, p<0.001) showed associations with the actual cognitive function (R-square =0.36). Associations with the HbA1c (R square = 0.37) showed the frequency of blood glucose self monitoring (beta = 0.38, p<0.001), the blood glucose value at the time of examination (beta = 0.34, p<0.001) and the diabetes duration (beta = 0.16, p = 0.03). There were no associations between actual cognitive function and other parameters. CONCLUSIONS: The data from this trial presents evidence which shows that treatment by general practitioners can enable patients both with and without insulin therapy to achieve a high quality of diabetes control. Hence, the results derived from ambulatory evaluations of treatment programmes can not be automatically tansferred to patients treated at hospitals. Regarding inpatient treatment, adapted structured treatment and teaching programmes are mandatory. PMID- 14555299 TI - Salmeterol versus sodium cromoglycate for the protection of exercise induced asthma in children--a randomised cross-over study. AB - OBJECTIVE: Physical activity is an important and frequent trigger of airways obstruction in asthmatic children. We aimed to compare the efficacies of 50 microg salmeterol twice daily and 2 mg SCG four times daily with respect to protection from exercise induced bronchoconstriction (EIB). METHODS: Twenty seven children and adolescents aged 4 to 16 years with mild or moderate exercise induced asthma (FEV1 70% to 90% predicted) were admitted to the study. Exercise challenge was performed on a treadmill using a predefined protocol in order to produce 10 minutes of exercise at near-maximum targets. The trial had a randomised, cross-over design comprising a 3-day run-in period and two 7-day treatment periods, separated by a one-week washout period. RESULTS: The mean protective efficacy of salmeterol was larger than that of SCG. A difference between treatments of 39.7% (95% CI, - 0,8 to 68.9%) in favour of salmeterol was calculated using a Hodges-Lehmann-estimate. The maximum post-challenge fall in FEV subset 1 was significantly lower (p<0.001) after salmeterol than after SCG (- 5.6 +/- 6.4% vs. -12.1 +/- 9.3%, respectively). In addition, salmeterol improved base-line lung function to a greater degree than SCG. FEV1 increased by 0.4 l/sec after salmeterol, whereas no improvement was observed after SCG. CONCLUSIONS: A one-week treatment with salmeterol 50 microg b.i.d in asthmatic children and adolescents provided better protection against EIB and improved baseline lung function as compared to SCG four times daily. PMID- 14555300 TI - Idiopathic thrombocytopenic purpura: an old disease revisited in the era of evidence-based medicine. PMID- 14555301 TI - Helicobacter pylori infection and immune thrombocytopenic purpura. PMID- 14555303 TI - Interaction of hemoglobin E and several forms of alpha-thalassemia in Cambodian families. AB - BACKGROUND AND OBJECTIVES: This study aimed to describe hematologic and molecular characterization of the interaction of hemoglobin (Hb) E and several forms of alpha-thalassemia causing complex thalassemia syndromes in two Cambodian families as well as to establish a rapid polymerase chain reaction (PCR) assay for simultaneous detection of Hb Constant Spring (CS) and Hb Pakse' (PS). DESIGN AND METHODS: Using PCR and DNA sequencing, the alpha- and beta-globin genotypes were examined. Clinical and hematologic data were assessed. A multiplex asymmetric allele-specific PCR for differential diagnosis of HbCS and HbPS was developed and validated. RESULTS: Eight genotypes including heterozygous HbCS, heterozygous HbPS, double heterozygous HbE/HbPS, double heterozygous HbE/alpha-thalassemia 2, triple heterozygous HbE/alpha-thalassemia /HbPS, homozygous HbE/alpha-thalassemia 2, compound alpha-thalassemia 2/HbCS and a hitherto undescribed compound HbCS/HbPS were found in these two families. Genotype-phenotype relationships are discussed and successful application of a multiplex PCR system for differential diagnosis of HbCS and HbPS is described. INTERPRETATION AND CONCLUSIONS: The interaction of several globin gene abnormalities in Cambodian families emphasizes the high frequencies of thalassemia and hemoglobinopathies. Identification of HbPS suggests that this mutation might be common and underestimated among South east Asian populations. A simplified PCR assay for simultaneous detection of HbCS and HbPS would facilitate characterization of these genotypes in both the clinical setting and population screening programs in the region. PMID- 14555304 TI - Asymptomatic and mild beta-thalassemia in homozygotes and compound heterozygotes for the IVS2+1G-->A mutation: role of the beta-globin gene haplotype. AB - BACKGROUND AND OBJECTIVES: We report on two families in which the beta(0) thalassemia mutation IVS2+1G-->A occurs either in the homozygous or compound heterozygous condition with other beta-thalassemia determinants. In the first family the proband, homozygous for the IVS2+1 determinant, is asymptomatic and was detected by chance during a screening program for beta-thalassemia. In the second family, the proband is a 43-year old female with a very mild thalassemia intermedia due to compound heterozygosity for the IVS2+1G>A and IVS1+110G>A mutations. Her father was diagnosed as having a thalassemic disorder only during the family studies carried out because of the proband's condition. He is a compound heterozygote for the Sicilian type deltabeta(0)-thalassemia and the IVS2+1 mutation and has a normal level of hemoglobin. DESIGN AND METHODS: In both families, the heterozygous carriers of the IVS2+1G>A have unusually elevated levels of fetal hemoglobin (HbF), and the homozygotes showed 98% HbF, reflecting an increased production of well hemoglobinized F-cells not associated with a significant erythroid expansion. RESULTS: The high HbF levels co-segregate with the beta-thalassemia mutation; the size and structure of both pedigrees do not allow the contribution of unlinked genes to the elevated production of HbF to be assessed. INTERPRETATION AND CONCLUSIONS: We propose that the unusual phenotypes resulting from homozygosity and compound heterozygosity for IVS2+1 are, against the background of a polygenic quantitative control of HbF expression, principally due to elements, such as repetitive sequences or single nucleotide polymorphisms, within or closely linked to the beta-gene cluster. These are potentially implicated in chromatin environment modifications, and could, therefore, be responsible for sustained HbF synthesis during development. PMID- 14555305 TI - Thalassemia minor, the Gilbert mutation, and the risk of gallstones. AB - BACKGROUND AND OBJECTIVES: Gallstones are a frequent complication of hemolytic anemias. The association with the mutation of the A(TA)nTAA motif of the promoter of the bilirubin UDP-glucuronosyltransferase gene has also been reported to increase the risk of gallstones. We studied the prevalence of cholelithiasis in thalassemia minor and the role of the Gilbert mutation. DESIGN AND METHODS: A group of 143 women obligate carriers of beta-thalassemia, and a control group of 170 hematologically normal women were compared. In both groups serum bilirubin, total cholesterol, and alanine-aminotransferase were measured and analysis of the mutation of the UGT-1A gene was performed. On the same occasion the women underwent ultrasonography. RESULTS: Total and unconjugated bilirubin were significantly higher in beta-thalassemia heterozygotes. Carriers of thalassemia had a higher prevalence of gallstones (20.3% vs 10.6% OR=2.15). Among the control group, the prevalence of gallstones did not differ significantly in relation to UGT1-A1 genotype, while in women carriers of beta-thalassemia it increased in an allele dose-dependent fashion. As compared to the controls, the odds ratios for the development of gallstones in thalassemic women were 1.68 (95% C.I.: 0.70 4.03) for those who had the normal UGT1-A1 genotype [(TA)6/(TA)6], 2.31 (95% C.I.: 1.06-5.02) for heterozygote carriers of the mutated genotype [(TA)7/(TA)6] and 3.88 (95% C.I.: 1.31-11.55) for those homozygous for the mutated genotype [(TA)7/(TA)7]. INTERPRETATION AND CONCLUSIONS: Thalassemia minor represents a risk factor for cholelithiasis and the Gilbert mutation further increases this risk. This is an additional example of how two genotypes can interact and modify a phenotype. PMID- 14555306 TI - Identification of two novel mutations in the 5'-untranslated region of H-ferritin using denaturing high performance liquid chromatography scanning. AB - BACKGROUND AND OBJECTIVES: Hereditary hyperferritinemia cataract syndrome is caused by mutations of the iron responsive elements (IREs) of L-ferritin mRNA. These alter the IRE structure and determine L-ferritin upregulation. IREs are located in 5'untranslated regions (5'UTR) of ferritin mRNAs. L-ferritin 5'UTR has been extensively studied and up to 21 different mutations have been identified. Only one mutation has been reported for H-ferritin 5'UTR; this mutation modified IRE structure and was apparently associated with high serum ferritin levels and iron overload. DESIGN AND METHODS: To identify other mutations in H ferritin 5'UTR we developed a fast DNA scanning method based on denaturing high performance liquid chromatography (HPLC). Five artificial DNA mutants were produced in order to validate the analytical conditions of the system for the identification of all mutations by single runs at 68 degrees C. The system was used to screen 660 DNA samples from subjects with high serum ferritin levels. RESULTS: Two abnormal patterns were identified carrying the mutations C20G and G34T. Structural data and the analysis of ferritin levels in red blood cells suggest that these mutations do not affect the functionality of the IRE. INTERPRETATION AND CONCLUSIONS: This large and first population analysis indicates that mutations in the H-ferritin 5'UTR are rare and do not seem to contribute to hyperferritinemia or iron overload. PMID- 14555307 TI - Imatinib mesylate therapy of chronic phase chronic myeloid leukemia resistant or intolerant to interferon: results and prognostic factors for response and progression-free survival in 150 patients. AB - BACKGROUND AND OBJECTIVES: Imatinib mesylate has recently been shown to be highly effective in chronic-phase chronic myeloid leukemia (CML). The results of imatinib treatment in chronic-phase CML patients resistant or intolerant to interferon (IFN) and the factors predicting therapeutic response and progression free survival were analyzed. DESIGN AND METHODS: One hundred and fifty patients with chronic-phase CML resistant (n=111) or intolerant (n=39) to IFN were treated with imatinib. Prognostic factors for response and disease progression were assessed by multivariate analysis. RESULTS: The median time from diagnosis was 43 months (0.5-188), median IFN therapy 21.5 months (0.5-140) and median follow-up from starting imatinib 13.6 months (range: 3-23). Complete hematologic response was achieved in 96 of 97 patients. Complete, partial and minor cytogenetic responses were present in 44%, 22%, and 8% of patients at 12 months. Grade III-IV neutropenia, thrombocytopenia, and anemia developed in 33%, 16%, and 6% of patients, respectively. Sixty-five patients discontinued treatment for a median of 4 weeks (1-36) due to toxicity. The rate of progression-free survival (lack of accelerated/blastic phase with persistent response) was 89.2% (95% CI: 84-94.4) at 12 months and 80.2% (95% CI: 72.2-88.2) at 18 months. Platelets > 450x10(9)/L and treatment discontinuation > 4 weeks were associated with a lower rate of major (complete plus partial) cytogenetic response. Patients in Sokal's high-risk group and those who did not achieve a major cytogenetic response had significantly shorter progression-free survival. INTERPRETATION AND CONCLUSIONS: Imatinib is highly effective in chronic-phase CML patients resistant or intolerant to IFN, especially in those with normal platelet counts and in those not requiring prolonged treatment discontinuation due to neutropenia. PMID- 14555308 TI - Clinical utility of the absolute number of circulating CD34-positive cells in patients with chronic myeloproliferative disorders. AB - BACKGROUND AND OBJECTIVES: Flow cytometry enumeration of peripheral blood CD34 positive cells provides reliable measurements of circulating hematopoietic progenitors in humans. Since the absolute number of circulating CD34-positive cells has been previously found to be elevated in myelofibrosis with myeloid metaplasia (MMM), we prospectively studied the clinical utility of this parameter in the work-up of patients with chronic myeloproliferative disorders. DESIGN AND METHODS: Of the 248 consecutive patients enrolled in this study, 106 had polycythemia vera (PV), 90 essential thrombocythemia (ET), and 52 myelofibrosis with myeloid metaplasia (MMM). The study population included both newly diagnosed and established cases, and of these latter some patients were on cytoreductive treatment while others were chemotherapy naive. Both cross-sectional and longitudinal investigations were carried out. Flow cytometry enumeration of CD34 positive cells was performed using a single-platform assay. RESULTS: Median numbers and ranges of circulating CD34-positive cells were 2.3x10(6)/L (0-5) in 20 control subjects, 2.2x10(6)/L (0-14) in those with PV, 2.4x10(6)/L (0-14) in those with ET, and 114x10(6)/L (6-2,520) in MMM patients. Analysis of variance demonstrated that values were markedly higher in MMM patients than in the remaining groups, and counts did not appear to fluctuate over short periods of follow-up. In both cross-sectional and longitudinal investigations on patients at clinical onset and/or out of cytoreductive treatment, a CD34-positive count of > or = 15x10(6)/L was always associated with MMM, clearly indicating a disease related specificity. INTERPRETATION AND CONCLUSIONS: The absolute number of circulating CD34-positive cells is normal or anyhow lower than 15x10(6)/L in patients with uncomplicated PV or ET, whereas it is equal or above this cut-off in those with MMM, likely reflecting abnormal hematopoietic progenitor cell trafficking. Thus, enumeration of circulating CD34-positive cells may be useful in the work-up of patients with myeloproliferative disorders. PMID- 14555309 TI - Anagrelide-induced bone marrow changes during therapy of chronic myeloproliferative disorders with thrombocytosis. an immunohistochemical and morphometric study of sequential trephine biopsies. AB - BACKGROUND AND OBJECTIVES: Anagrelide is an agent with a significant platelet lowering activity in humans. Contrasting the wealth of clinical data, bone marrow (BM) changes during therapy have been relatively rarely studied; information is particularly lacking regarding specific features of megakaryocytopoiesis. DESIGN AND METHODS: A study was performed on 15 patients with essential thrombocythemia and early stage chronic idiopathic myelofibrosis presenting with an elevated platelet count. These patients received anagrelide for 25 months resulting in a significant improvement of thrombocytosis. Evaluations were carried out by morphometry on sequential BM biopsies following enzyme- and immunohistochemical stainings that also included those for proliferative capacity and apoptosis. RESULTS: No significant change in proliferation or apoptosis was recognizable during anagrelide treatment. The most conspicuous alterations were those of the CD61+ megakaryocytopoiesis. Megakaryocytopoiesis revealed an increase in promegakaryoblasts together with an enhancement of proliferating cell nuclear antigen activity. This feature is in keeping with an inhibitory effect on endoreduplication implying an arrest of polyploidization and maturation into platelet-shedding large megakaryocytes. On the other hand, a significant increase in the number of megakaryocytes was not detectable. Anagrelide failed to exert a stimulating influence on the progression of myelofibrosis or on the amount of CD34+ progenitor cells. Regarding angiogenesis, there was no increase in the density of BM vessels, but distension of the vascular lumina corresponded with a vasodilatory effect. INTERPRETATION AND CONCLUSIONS: Anagrelide exerts a significant effect on endoreduplicative activity of megakaryocytes consistent with an inhibition of maturation and therefore, generates a relative predominance of precursor cells but fails to stimulate myelofibrosis. PMID- 14555310 TI - Idiotype vaccination of multiple myeloma patients using monocyte-derived dendritic cells. AB - BACKGROUND AND OBJECTIVES: Dendritic cells (DC) pulsed with multiple myeloma (MM) patient-specific idiotype (Id) protein can induce MM-specific T-cell responses. DESIGN AND METHODS: We established serum-free culture conditions to generate monocyte-derived DC for clinical use to circumvent anti-xenogenic immune responses with repetitive vaccinations. In a clinical phase I trial twelve patients responsive to high dose chemotherapy (HDT) were vaccinated with autologous Id pulsed DC vaccines followed by Id/keyhole limpet hemocyanin (Id/KLH) booster immunizations co-injected with granulocyte-macrophage colony stimulating factor as adjuvant. RESULTS: In vitro studies showed that serum-free generated DC were equally effective in the induction of specific T-cell responses as were DC generated with fetal calf serum. On average 4.5 x 10(6) DC of >60% purity were generated from peripheral blood monocytes obtained 3-6 months after HDT and autologous stem cell transplantation. Ten of twelve patients received all planned vaccines without serious toxicity. Two patients developed Id-specific T cell proliferative responses, in one patient an Id-specific cytotoxic T lymphocyte (CTL) response was measured. Id-specific TH1 cytokine secretion was found in one of the T-cell responding patients. All patients who received at least three Id/KLH vaccines mounted strong KLH specific T-cell and delayed antibody responses. Two patients remain in clinical partial response at 25 and 29 months after the start of the vaccination and ten patients have progressed, six of whom have died from progressive disease or infectious complications. INTERPRETATION AND CONCLUSIONS: Serum-free DC vaccines induce Id-specific T-cell responses in MM patients. PMID- 14555311 TI - Endogenous factor V synthesis in megakaryocytes contributes negligibly to the platelet factor V pool. AB - BACKGROUND AND OBJECTIVES: Coagulation factor V (FV) is distributed between two pools: 80% circulates in plasma and 20% is stored in platelets. The aim of the study was to determine the origin of platelet FV. DESIGN AND METHODS: We investigated a FV Leiden heterozygous patient who had received an allogeneic bone marrow transplant from a normal donor. The patient had been referred to our laboratory for his marked activated protein C (APC) resistance in the apparent absence of FV Leiden. Analysis of the DNA from a buccal swab showed that the patient was indeed a heterozygous carrier of FV Leiden. The difference in FV genotype between the hepatocytes (heterozygous FV Leiden) and the blood cells (homozygous normal) of the patient provided a good model to investigate the origin of platelet FV. Platelets were isolated from the patient and the bone marrow donor and activated with thrombin and ionomycin to release and activate FV. APC was then added and the inactivation of platelet FVa was followed over time with a highly sensitive prothrombinase-based assay. RESULTS: While the donor's platelet FVa showed a normal inactivation time course, the patient's platelet FVa was considerably resistant to APC. The kinetic pattern of APC catalyzed inactivation of the patient's platelet FVa was indistinguishable from that of plasma FVa from a FV Leiden heterozygote. INTERPRETATION AND CONCLUSIONS: These data indicate that platelet FV is derived from plasma and that endogenous FV synthesis by megakaryocytes contributes negligibly to the platelet FV pool. PMID- 14555312 TI - Comparison of six-month outcome of patients initially treated for acute deep vein thrombosis with a low molecular weight heparin Certoparin at a fixed, body-weight independent dosage or unfractionated heparin. AB - BACKGROUND AND OBJECTIVES: Body weight-adjusted subcutaneous low molecular weight heparin (LMWH) has been proven to be more effective and safer than aPTT-adjusted intravenous unfractionated heparin (UFH) for the initial treatment of patients with acute symptomatic deep venous thrombosis (DVT) based on analyses pooling the results of studies with different LMWHs. We investigated whether these findings hold for a particular LMWH by pooling the results of two independent studies. DESIGN AND METHODS: Patients with acute symptomatic proximal DVT (n=1758), proven by ascending phlebography or compression ultrasound, received either a fixed, body weight independent dose of 8,000 IU Certoparin b.i.d. (n=893) for 8.6 days or intravenous UFH (n=865) adjusted to an 1.5 to 3.0-fold prolongation of the aPTT for 12.0 days both followed by vitamin K-antagonists for 6 months. RESULTS: Venous thromboembolism (VTE) re-occurred in 5.1% and 3.1% (RRR 0.62, CI 0.39 0.98, 2p=0.04), major bleeding in 3.5% and 1.9% (RRR 0.55, CI 0.31-0.99, 2p=0.05), mortality in 3.6% and 2.1% (RRR 0.59, CI 0.34-1.04, 2p=0.08), and the composite outcome of all three events in 10.3% and 6.3% (RRR 0.61, CI 0.44 to 0.84, 2p=0.002) of patients at 6 months initially randomised to UFH and LMWH, respectively. INTERPRETATION AND CONCLUSIONS: The initial treatment of acute DVT with a fixed dose of the LMWH, certoparin, is more effective in reducing, over 6 months, the re-occurrence of VTE and the composite outcome of recurrent VTE, major bleeding, and mortality without any relation of the bodyweight of the patients to recurrent venous thromboembolism or major bleeding complications. PMID- 14555313 TI - Female genital tract graft-versus-host disease following allogeneic bone marrow transplantation. AB - BACKGROUND AND OBJECTIVES: Graft-versus-host disease (GVHD) is a complex syndrome observed after bone marrow transplantation (BMT) affecting several organs including the lower genital female tract. We tried to evaluate the incidence of genital tract involvement and whether there are specific risk factors. DESIGN AND METHODS: A retrospective study was conducted in order to describe genital manifestations of GVHD and evaluate its incidence, severity and remission among 213 females who underwent BMT. The risk factors studied were previous pregnancies, vaginal cultures just before BMT and hormonal replacement therapy (HRT). RESULTS: Genital lesions considered as expression of GVHD were found in 53 patients (24.9%). They appeared in the first 100 days after BMT in 12 women and beyond in 41 cases. Seventy-three percent of patients with such lesions showed some evidence of chronic GVHD elsewhere. The proposed grading, the first attempt of its kind, showed that genital chronic GVHD was minimal in 66%, moderate in 22% and severe in 12% of patients. Vaginal fibrosis, sometimes with complete obstruction, was seen in this last form. This occurred in 86.8% of patients after 2-157 months (median 22) while persistent GVHD was observed in 7 of them. In our sample no significant association was found between genital GVHD and previous pregnancies or vaginal infections at BMT, while HRT seems poorly associated with gynecological manifestations of GVHD (p=0.049). INTERPRETATION AND CONCLUSIONS: Genital GVHD is not unusual after BMT. It can seriously affect female sexuality and the overall quality of life. We suggest stressing the importance of early detection of genital involvement in order to prevent the most serious lesions. Further studies are needed to identify the triggering factors associated with the development of genital GVHD. PMID- 14555314 TI - Prospects for ultraviolet A1 phototherapy as a treatment for chronic cutaneous graft-versus-host disease. AB - BACKGROUND AND OBJECTIVES: Standard or investigative immunosuppressive therapies for cutaneous chronic graft-versus-host disease (GVHD) may prove not only ineffective but also cause serious adverse effects. Repeated exposure of the skin to ultraviolet radiation in the wavelength range 340-400 nm (so-called ultraviolet A1) was recently reported to have a strong local (intracutaneous) immunomodulatory activity. This study was undertaken to evaluate efficacy and safety of this phototherapy. DESIGN AND METHODS: Nine patients with cutaneous (4 lichenoid and 5 sclerodermoid) GVHD and mild or no other organ involvement were enrolled. All patients had developed serious drug toxicity and/or opportunistic infections. Phototherapy was administered three times a week. RESULTS: Complete remission was seen in 5 (2 lichenoid and 3 sclerodermoid) cases and a partial improvement in 4 (2 lichenoid and 2 sclerodermoid) after having received 15.8+/ 3.8 (lichenoid GVHD) or 21.6+/-8.0 (sclerodermoid GVHD) sessions of phototherapy. Adverse effects were not registered. At follow-up (range: 6-25 months), two patients with sclerodermoid lesions relapsed after 5 months but responded to another treatment cycle. Patients with lichenoid GVHD showed relapses within one month and prolonged maintenance phototherapy was needed. Problems of drug toxicity and opportunistic infections improved as phototherapy allowed the reduction or interruption of systemic drug therapies. INTERPRETATION AND CONCLUSIONS: Ultraviolet A1 phototherapy may be considered as an appropriate therapeutic approach for sclerodermoid GVHD with no or mild involvement of internal organs. Patients with lichenoid GVHD should be treated only if they develop serious adverse effects to immunosuppressive therapies and opportunistic infections because of the carcinogenic hazard of high cumulative doses of ultraviolet A1 radiation. PMID- 14555315 TI - Staging Hodgkin's lymphoma: why and how? PMID- 14555316 TI - The factor V HR2 haplotype and the risk of venous thrombosis: a meta-analysis. AB - BACKGROUND AND OBJECTIVES: A complex haplotype of factor V gene (FV HR2) has been recently reported. FVHR2 possesses decreased co-factor activity to APC in the degradation of FVIIIa, and an increased ratio of the more procoagulant isoform FV1 compared to FV2. Contrasting results on whether the haplotype induces a significant risk of venous thromboembolism (VTE) have been reported. DESIGN AND METHODS: It has been surmised that FVHR2 enhances the risk of VTE carried by FV Leiden. We carried out a meta-analysis of the reported studies on the role of HR2 haplotype in inducing a risk of VTE and the influence of the polymorphism on the risk carried by patients with FV Leiden. RESULTS: Eight studies were analyzed for the estimation of the risk of VTE. A total of 338 out of 2,696 cases (12.5%; range 7.8 to 18.5%) and 885 out of 7,710 controls (11.5%; range 8.1 to 12.1%) were HR2 positive. The odds ratio for VTE associated with HR2 haplotype was not statistically significant (OR 1.15; 95% C.I. 0.98-1.36). The OR for the association between FV Leiden and FV HR2 and the risk of VTE in cases and controls was largely heterogeneous as to OR and 95% C.I. and no statistical significant difference was observed. INTERPRETATION AND CONCLUSIONS: The data from the present meta-analysis suggests that FVHR2 could be a very mild prothrombotic factor. The association of FV Leiden and HR2 haplotype seems not to increase significantly the risk of VTE carried by isolated heterozygosity for FV Leiden. However, well-designed clinical studies are needed to clarify this issue definitely. PMID- 14555317 TI - Familial chronic lymphocytic leukemia. AB - Familial aggregation of chronic lymphocytic leukemia (CLL) has been observed more frequently than familial aggregation of any other type of oncohematologic disorder. The presence of cells with a CLL-like immunophenotype (CLL-like cells) was recently documented in 13.5% healthy first-degree relatives of CLL patients. We present a family with CLL in which 2 brothers, a sister and their mother were affected. PMID- 14555318 TI - A rare beta-thalassaemia mutation (C-T) at position -90 of the beta-globin gene discovered in a Chinese family. AB - We provide the first description of a Chinese family with three heterozygotes for a rare beta-thalassemia mutation previously observed in a Portuguese carrier. The mutation (-90 C-T) changes the conserved promoter sequence within the proximal CACCC box of the beta-globin gene; this can reduce beta-globin transcription significantly. PMID- 14555319 TI - Gilbert's syndrome as a predisposing factor for idiopathic cholelithiasis in children. AB - The frequency of the (TA)7/(TA)7 promoter genotype of UDP-glucuronosyltransferase gene (UGT1A1) was significantly higher (p<0.05) in a group of 30 children with cholelithiasis than in a control group of 40 healthy children, indicating that this genotype might be an underlying factor for gallstone initiation in otherwise healthy children. PMID- 14555320 TI - Immunological reconstitution in long-term survivors of thalassemia major patients after hematopoietic stem cell transplantation. AB - We studied the immune function of 33 long-term survivors of thalassemia after hematopoietic stem cell transplantation. Lymphocyte subsets, lymphoproliferative response and immunoglobulin were normal but the level of natural killer cells was low. Five and seven patients had suboptimal antibody response at 4 week after pneumococcal and hepatitis B vaccine, respectively, but this response returned to normal by 6 months. PMID- 14555321 TI - alpha-globin gene deletion and point mutation analysis among in Iranian patients with microcytic hypochromic anemia. AB - We tested 67 Iranian individuals, presenting with low mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) levels, normal hemoglobin electrophoresis and iron status, for the presence of twelve common alpha thalassemia gene deletions and point mutations. Five different mutations ( alpha(3.7), -alpha(4.2), --MED, -(alpha)20.5, Hb Constant Spring) were identified in a total of 43 cases PMID- 14555322 TI - Incidence and characteristics of myelodysplastic syndromes in Ourense (Spain) between 1994-1998. AB - The very few reference epidemiological studies on myelodysplastic syndromes (MDS) have been carried out in Europe: Germany, France, UK and Sweden. We present the first Spanish study on the incidence and characteristics of MDS. The incidence rates, distribution by FAB subtypes, sex and age groups are within the ranges established by the reference studies with minimal differences which we point out and attempt to explain. PMID- 14555323 TI - Safety and efficacy of stem cell mobilization under imatinib therapy. AB - In order to investigate the safety and efficacy of stem cell mobilization in chronic myeloid leukemia patients under imatinib therapy we treated 10 such patients with granulocyte colony-stimulating factor. We observed that none of the patients developed progressive disease under this treatment. Instead, sufficient CD34+ apheresis could be performed in 7 patients and, as assessed by nested reverse transcriptase polymerase chain reaction (RT-PCR), bcr/abl-negative stem cell products could be generated in 3 patients. Interestingly, in 3 other patients with bcr/abl-positivity in 1st round RT-PCR of peripheral leukocytes, bcr/abl transcripts in stem cell products could only be detected by nested RT PCR. PMID- 14555324 TI - Induction of apoptosis by castration in epithelium of the mouse seminal vesicles. AB - Castration on days 0, 5, 10, 20, 40, and 60 caused increases in an apoptotic index (% of apoptotic cells) in seminal vesicle (SV) epithelium, peaking 1-3 days after castration. The peak apoptotic indices after castration on days 0, 5, 10, and 20 were significantly lower than peak apoptotic indices observed after castration on days 40 and 60. DNA extracted from mouse SVs 2 days after castration on days 0, 5, 10, and 60 showed a ladder pattern on agarose gel electrophoresis. The secretion of androgen by testes was confirmed by the growth retardation of the SVs after castration on days 0, 5, 10, and 20. It would appear that a proportion of SV epithelial cells dependent on testicular androgens for survival is smaller before day 20 than after day 20. PMID- 14555325 TI - Transurethral incision for hematospermia caused by ejaculatory duct obstruction. AB - Hematospermia is supposed to be derived from pathological conditions in the seminal vesicle, prostate, testis, epididymis, or urethra. A recent advance in diagnostic procedures has demonstrated the seminal vesicle, the prostate, and midline cyst as potential sources of hematospermia. The authors describe a case of hematospermia caused by ejaculatory duct obstruction, in which a transurethral technique was successful. A 51-year-old male was referred to the authors' clinic with a chief complaint of hematospermia. Transurethral ultrasonography showed a cystic lesion surrounded with hyperechoic area in the middle of the prostate. Vasography demonstrated the distal dilation of the ejaculatory duct. Magnetic resonance imaging demonstrated a high signal intensity area in the middle of the prostate. Urethrocystoscopy showed an enlarged cystic lesion with an orifice at the prostatic urethra, which was incised endoscopically. There was no complication observed postoperatively. Seven months after the technique, hematospermia resolved completely. A midline cyst should be considered a cause of hematospermia and the incidence of such cysts may be higher than that previously recognized. The transurethral technique is expected to be a successful treatment approach. PMID- 14555326 TI - Expression of Bcl-2 and Bax in rhesus monkey testis during germ cell apoptosis induced by testosterone undecanoate. AB - Apoptosis occurs spontaneously during spermatogenesis and can be induced by androgen withdrawal. However, the molecular events governing apoptosis have not been characterized. To study the molecular mechanism of apoptosis induced by a high dose of testosterone undecanoate (TU), the authors examined the temporal changes in proapoptotic Bax and antiapoptotic Bcl-2 in TU-treated monkey testes. Apoptotic cells were identified in tissue sections by in situ end labeling of fragmented DNA. The results showed that a great deal of the apoptotic cells occurred in the testes on day 30 after TU injection and that the dominant apoptotic germ cells are spermatocytes and spermatids. The expression of Bcl-2 and Bax was assessed by immunohistochemical method and Western blot. As compared with that of normal testes, the levels of Bcl-2 protein increased significantly from 7 to day 14 while that of Bax protein was almost unchanged in the testes from day 7 up to day 60 after TU treatment. Bcl-2 was localized to the spermatids in the normal testes and temporarily distributed in both the cytoplasm and nucleus of those cell types susceptible to TU-induced apoptosis on day 14 after TU injection. Therefore, it is suggested that Bax may not play a role in initiating germ cell apoptosis induced by TU injection and that the evaluation in Bcl-2 expression may represent a survival mechanism for the remaining germ cell. PMID- 14555327 TI - Soluble Fas and interleukin-6 and interleukin-8 levels in seminal plasma of infertile men. AB - The seminal plasma levels of soluble Fas (sFas) and interleukins IL-6 and IL-8 were measured and their relationship with semen qualities was examined. The seminal plasma sFas level in fertile males was significantly higher than that in the infertile group. On the other hand, seminal plasma level of IL-6 was significantly lower in fertile males than in the infertile group. In the infertile group, patients with oligozoospermia had a lower seminal plasma sFas and a higher IL-6 level than those with normal sperm concentration. There was an inverse correlation between IL-6 and sperm concentration in infertile patients. Seminal plasma IL-8 was not correlated with sperm parameters. It would appear that certain kinds of cytokine in the seminal plasma might play an important role in improving semen quality. PMID- 14555328 TI - Hematuria from left internal pudendal and obturator arterial bleeding following sexual intercourse. AB - Bladder tamponade is an uncommon clinical symptom among men who experience suffering related to sexual intercourse. The authors report on a 46-year-old man with this symptom 4-5 years before hospitalization. Angiography confirmed the presence of left pudendal and obturator arterial bleeding, and embolotherapy of the internal pudendal and obturator arteries was performed. There was no mortality, or limb loss or loss of sexual potency at follow-up. PMID- 14555329 TI - Angiontensin-converting enzyme activity in dunning rat prostate tumor. AB - A dipeptidyl carboxypeptidase activity in the Dunning rat prostate tumor was characterized. This enzyme demonstrated the most prominent properties of angiotensin converting enzyme (ACE): that is, it was stimulated by NaCl and Co(2+) and was potently inhibited by captopril. The enzyme solubilized by Triton X-100 had a molecular mass of 110 kDa as determined by gel filtration chromatography. The specific activity of ACE did not change with castration, indicating that ACE activities are not controlled by androgen. The role of ACE in the prostate and its tumors is not understood, but the ability of this enzyme to hydrolyze a number of bioactive peptides suggests that it may function in controlling the molecular forms or activity of regulatory peptides. PMID- 14555330 TI - Povidone-iodine instillation for management of pelvic lymphocele after pelvic lymphadenectomy for staging prostate cancer. AB - The development of postoperative lymphoceles following pelvic lymph node dissection is a rare complication. It is a well-described complication of kidney transplantation. A patient who developed a symptomatic pelvic lymphocele after pelvic lymph node dissection for staging prostatic cancer was treated with percutaneous tube drainage, but the treatment was in vain. Successful treatment was accomplished with povidone-iodine instillation into the lymphocele. This simple, safe, and painless method for lymphocele treatment is recommended. PMID- 14555332 TI - Value of prostate-specific antigen in the staging of Taiwanese patients with newly diagnosed prostate cancer. AB - Records of 71 patients diagnosed with prostate cancer were reviewed retrospectively regarding clinical stage, prostate-specific antigen (PSA), Gleason score, CT scan of pelvis, bone scan, and pelvic lymph node dissection. Fourteen patients had pelvic lymphadenopathy based on the CT scan. Of these, no patient had a PSA level <4 ng/mL, 1 patient had a PSA level between 4 and 10 ng/mL, and 3 had a PSA level between 10 and 20 ng/mL. Twelve of 13 patients with positive bone scan results had a PSA level >20 ng/mL, and 1 patient had a PSA level between 10 and 20 ng/mL. PSA can be cost-effective in selecting and identifying appropriate staging for patients with newly diagnosed prostate cancer. CT scans are not indicated in men with clinical localized prostate cancer when PSA levels are < or =10 ng/mL. Bone scan is not required for staging asymptomatic men with PSA levels of < or =20 ng/mL. Pelvic lymphadenectomy for localized prostate cancer may not be necessary if PSA levels is < or =20 ng/mL and Gleason score is < or =5. PMID- 14555331 TI - Primary B cell lymphoma of the penis: successful treatment with organ preservation. AB - Malignant lymphoma uncommonly manifests in the genitourinary tract. Primary penile lymphoma is extremely rare. A 77-year-old male presented with primary malignant B-cell lymphoma of the penis with the chief complaint of a painless and itching nodule on the penile glans for more than 2 months. The pathologic examination with immunohistochemical stain of penile biopsy revealed malignant B cell lymphoma, mixed cellular type. The physical examination and the computed tomography scan of chest, abdomen, and pelvis showed no evidence of superficial, thoracic, abdominal, or pelvic lymphadenopathy. This case was treated with local excision and systemic chemotherapy with good cosmetic and functional results. There was absence of recurrence 16 months after therapy. PMID- 14555333 TI - Indication of varicocelectomy in the era of assisted reproductive technology: prediction of treatment outcome by noninvasive diagnostic methods. AB - Of 103 patients who underwent varicocelectomies, 87 underwent all the diagnostic methods: color Doppler ultrasonography (DOP), scrotal deep body temperature (DBT), and scrotal scintigraphy (SSG). Total motile sperm (TM) was calculated. The positivities of DOP, DBT, and SSG were 67, 51, and 90%, respectively. In the patients <30 years old, TM increased in the DBT-positive group. In older patients, positive for DBT indicated poor improvement of TM, whereas positive for DOP indicated better response in the group. Patients older than 30 years and positive for DBT, but not DOP, should be counseled directly to receive intrauterine insemination or in vitro fertilization/intracytoplasmic sperm injection. PMID- 14555334 TI - Persistent Mullerian duct syndrome: a surgical approach. AB - Laparoscopy revealed a left inguinal testis and a right abdominal testis. Surgery revealed uterus-like structures. The bilateral testes showed primitive testis without ovarian tissue. Physical examination showed a normal and an empty scrotum with a nonpalpable gonad. Chromosome analysis revealed 46,XY. Pathological findings demonstrated the immature testis and the immature uterus. PMID- 14555336 TI - Identification of cytochrome P450 enzymes involved in the metabolism of 4' methoxy-alpha-pyrrolidinopropiophenone (MOPPP), a designer drug, in human liver microsomes. AB - 1. The metabolism of 4'-methoxy-alpha-pyrrolidinopropiophenone (MOPPP), a novel designer drug, to its demethylated major metabolite 4'-hydroxy-pyrrolidinopropio phenone (HO-PPP) was studied in pooled human liver microsomes (HLM) and in cDNA expressed human hepatic cytochrome P450 (CYP) enzymes. 2. CYP2C19 catalysed the demethylation with apparent Km and Vmax values of 373.4 +/- 45.1 microM and 6.0 +/- 0.3 pmol min(-1) pmol(-1) CYP, respectively (mean +/- SD). Both CYP2D6 and HLM exhibited clear biphasic profiles with apparent K(m,1) values of 1.3 +/- 0.4 and 22.0 +/- 6.5 microM, respectively, and V(max,1) values of 1.1 +/- 0.1 pmol min(-1) pmol(-1) CYP and 169.1 +/- 20.5 pmol min(-1) mg(-1) protein, respectively. 3. Percentages of intrinsic clearances of MOPPP by particular CYPs were calculated using the relative activity factor (RAF) approach with (S) mephenytoin-4'-hydroxylation or bufuralol-1'-hydroxylation as index reactions for CYP2C19 or CYP2D6, respectively. 4. MOPPP, HO-PPP and the standard 3',4' methylenedioxy-pyrrolidinopropio-phenone (MDPPP) were separated and analysed by liquid chromatography-mass spectrometry in the selected-ion monitoring (SIM) mode. 5. The CYP2D6 specific chemical inhibitor quinidine (3 microM) significantly (p<0.0001) inhibited HO-PPP formation by 91.8 +/- 0.5% (mean +/- SEM) in incubation mixtures with HLM and 2 microM MOPPP. 6. It can be concluded from the data obtained from kinetic and inhibition studies that polymorphically expressed CYP2D6 is the enzyme mainly responsible for MOPPP demethylation. PMID- 14555335 TI - Mechanism-based inhibition of CYP activities in rat liver by fluoxetine and structurally similar alkylamines. AB - 1. The inhibition of cytochrome P450 (CYP)-mediated substrate oxidations by alkylamine-based drugs was investigated in rat hepatic microsomes. The effects of pre-incubation of the drugs with NADPH-fortified microsomes on inhibition potency was evaluated in relation to the formation of metabolite intermediate (MI) complexes with CYP in vitro. 2. The selective serotonin-reuptake inhibitor fluoxetine (FLU) emerged as a potent and preferential inhibitor of CYP2C11 in rat liver microsomes. After FLU biotransformation in NADPH-supplemented microsomes, IC50 values of 2 and 1 microM were determined against CYP2C11-dependent testosterone 2alpha- and 16alpha-hydroxylation; in the absence of pre-incubation, the corresponding IC50 values were 47 and 39 microM. MI complexation of CYP appeared to contribute significantly to inhibition by FLU, as evidenced by the 21% decrease in apparent microsomal CYP content produced by 50 microM FLU in the presence of NADPH. 3. The secondary amines nisoxetine (NIS), and especially, desipramine (DES) and nortriptyline (NOR), also inhibited CYP2C11 and generated MI complexes with microsomal CYP. In contrast, with the exception of SKF-525-A, tertiary alkylamines (10 compounds) inhibited specific CYP activities but did not form MI complexes. Pre-incubation of these agents with NADPH-supplemented microsomes did not enhance inhibition of CYP activities, thus suggesting that formation of inhibitory metabolites was minimal for these compounds. 4. These findings implicate drug-mediated MI complexation of CYPs in the inhibition of hepatic biotransformation processes by secondary alkylamines. In contrast, tertiary amines did not generate significant quantities of CYP-MI complexes under the test conditions. Despite their diffusion from the CYP active site, inhibition produced by tertiary amines and stable metabolites of other drugs may be significant. However, such inhibition would be of shorter duration than that from MI complexation, which involves quasi-covalent binding to the haem and prevention of oxygen activation. PMID- 14555337 TI - Characterization of bropirimine O-glucuronidation in human liver microsomes. AB - 1. The antitumour agent bropirimine undergoes significant Phase II conjugation in vivo. Incubation of [14C]bropirimine with human liver microsomes resulted in the formation of a single product peak (M1) using high-performance liquid chromatography with radiochemical detection and was tentatively assigned as bropirimine glucuronide based on sensitivity to beta-glucuronidase and by obtaining the expected mass of 442/444 amu with liquid chromatography/mass spectrometry. Following metabolite isolation, the structure of M1 was established as bropirimine O-glucuronide by 1H-nuclear magnetic spectroscopy. 2. Studies aimed at identifying the human liver UDP-glucuronosyltransferase (UGT) enzyme(s) involved in the glucuronidation of bropirimine were carried out using recombinant human UGTs and it was determined that glucuronidation of bropirimine was catalysed by UGT1A1, UGT1A3 and UGT1A9. Bropirimine O-glucuronidation followed Michaelis-Menten kinetics and the Km and Vmax (mean +/- SD; n = 3) were 1217 +/- 205 microM and 667 +/- 188 pmol min(-1) mg(-1), respectively. 3. The activity of bropirimine O-glucuronidation by human liver microsomes was inhibited by bilirubin (40%) and with mefenamic acid (80%). Although buprenorphine extensively inhibited the activity of bropirimine O-glucuronidation by UGT1A3, the inhibition profile did not parallel that observed in HLMs. 4. The results demonstrate that UGT1A9 and to a lesser extent UGT1A1 are responsible for the majority of bropirimine O-glucuronidation in man. PMID- 14555338 TI - Identification of some N-hydroxylated metabolites of (+/-)-3,4 methylenedioxymethamphetamine in horse urine by gas chromatography-mass spectrometry. AB - 1. The in vivo biotransformation of (+/-)-3,4-methylenedioxymethamphetamine [(+/ )-MDMA] in the thoroughbred horse was determined after oral administration. 2. Unconjugated compounds and aglycones were isolated from enzyme-hydrolysed urine by solid-phase extraction using mixed-mode cartridges. The basic isolates were derivatized (trimethylsilylether, TMS) and analysed by positive-ion electron ionization/gas chromatography-mass spectrometry (EI+/GC-MS). MDMA and 10 Phase I metabolites containing the arylisopropylamine substructure were detected. 3. N Hydroxy amphetamine and N-hydroxymethamphetamine were synthesized. The EI + mass spectra of their O-TMS derivatives showed characteristic alpha-cleavage ions at m/z 132 and 146, respectively, as base peaks. Based upon these data, five putative N-hydroxylated metabolites of MDMA were detected. 4. In the horse, (+/-) MDMA is metabolized by oxidative N-demethylation to form the primary amine methylenedioxyamphetamine (MDA). Both MDMA and MDA are further metabolized by oxidative demethylenation (cleavage and O-demethylation of the benzodioxole moiety) to form the corresponding catechols, 3-O-methylation to form the guaiacols and N-oxidation of the secondary and primary amine metabolites to form the hydroxylamines. 5. Both phenolic and N-hydroxy metabolites of (+/-)-MDMA undergo Phase II conjugation before excretion in urine. PMID- 14555339 TI - Metabolism and disposition of gemfibrozil in Wistar and multidrug resistance associated protein 2-deficient TR- rats. AB - 1. The roles of multidrug resistance-associated protein (Mrp) 2 deficiency and Mrp3 up-regulation were evaluated on the metabolism and disposition of gemfibrozil. 2. Results from in vitro studies in microsomes showed that the hepatic intrinsic clearance (CLint) for the oxidative metabolism of gemfibrozil was slightly higher (1.5-fold) in male TR- rats, which are deficient in Mrp2, than in wild-type Wistar rats, whereas CLint for glucuronidation was similar in both strains. 3. The biliary excretion of intravenously administered [14C]gemfibrozil was significantly impaired in TR-) rats compared with Wistar rats (22 versus 93% of the dose excreted as the acyl glucuronides over 72 h). Additionally, the extent of urinary excretion of radioactivity was much higher in TR- than in Wistar rats (78 versus 2.6% of the dose). 4. There were complex time dependent changes in the total radioactivity levels and metabolite profiles in plasma, liver and kidney, some of which appeared to be related to the up regulation of Mrp3. 5. Overall, it was demonstrated that alterations in the expression of the transporters Mrp2 and Mrp3 significantly affected the excretion as well as the secondary metabolism and distribution of [14C]gemfibrozil. PMID- 14555340 TI - Disposition of flurbiprofen in man: influence of stereochemistry and age. AB - 1. The stereoselective metabolism and pharmacokinetics of the enantiomers of flurbiprofen were investigated following the oral administration of the racemic drug (100 mg) to four young and four elderly healthy volunteers (two males and two females per group). 2. The stereochemical composition of the drug and the 4' hydroxy- metabolite in serum and the drug, 4'-hydroxy- and 3'-hydroxy-4'-methoxy- metabolites, both free and conjugated, in urine were determined by a direct chromatographic method of enantiomeric analysis. 3. Modest enantioselectivity in clearance (CL S/R: young, 0.86; elderly, 0.88) was largely responsible for the apparent elimination half-life of (S)-flurbiprofen being significantly greater (p<0.01) than that of the R-enantiomer in both age groups (young, S: 5.2 +/- 0.7 versus R: 4.5 +/- 0.6 h; elderly, S: 9.6 +/- 1.2 versus R: 7.1 +/- 1.0 h). The serum concentrations of 4'-hydroxyflurbiprofen were five- to 20-fold lower than those of the corresponding drug enantiomers, stereoselective disposition being evident in the significantly greater (p<0.05) apparent half-lives of the S- compared with the R-enantiomer in both groups (young, S: 10.6 +/- 2.4 versus R: 6.7 +/- 1.1 h; elderly, S: 13.7 +/- 1.7 versus R: 10.2 +/- 1.2 h). 4. Some 60 and 72% of the dose was excreted in 24-h urine in elderly and young volunteers, respectively, a significantly greater (p<0.05) proportion of which was of the R configuration in both age groups (S/R: young, 0.87; elderly, 0.81). The major urinary excretion products were flurbiprofen and 4'-hydroxyflurbiprofen, and their acyl-conjugates in both groups. 5. Age-associated differences in the pharmacokinetics of flurbiprofen occurred in a non-stereoselective manner and were primarily as a consequence of a significant approximately 40% decrease (p<0.01) in clearance of both enantiomers in the elderly due to reduced metabolic activity. Consequently, the elderly had greater exposure to both enantiomers, as reflected by the AUCs(0-inf) being significantly higher (p<0.05), by 60%, in this age group compared with the young. 6. The findings suggest that age-related alterations in the disposition of flurbiprofen could have significant implications for the use of the drug in the elderly. PMID- 14555341 TI - Oral absorption, metabolism and excretion of 1-phenoxy-2-propanol in rats. AB - 1. This study was designed to determine the absorption, metabolism and excretion of 1-phenoxy-2-propanol in Fischer 344 rats following oral administration in an effort to bridge data with other propylene glycol ethers. 2. Rats were administered a single oral dose of 10 or 100 mg kg(-1) 14C-1-phenoxy-2-propanol as a suspension in 0.5% methyl cellulose ether in water (w/w). Urine was collected at 0-12, 12-24 and 24-48 h and faeces at 0-24 and 24-48 h post-dosing and the radioactivity was determined. Urine samples were pooled by time point and dose level and analysed for metabolites using LC/ESI/MS and LC/ESI/MS/MS. 3. The administered doses were rapidly absorbed from the gastrointestinal tract and excreted. The major route of excretion was via the urine, accounting for 93 +/- 5% of the low and 96 +/- 3% of the high dose. Most of the urinary excretion of radioactivity occurred within 12 h after dosing; 85 +/- 2% of the low and 90 +/- 1% of the high dose. Total faecal excretion remained < 10%. Rats eliminated the entire administered dose within 48 h after dosing; recovery of the administered dose ranged from 100 to 106%. Metabolites tentatively identified in urine were conjugates of phenol (sulphate, glutathione) with very low levels (< 2%) of hydroquinone (glucuronide), conjugates of parent compound (glucuronide, sulphate) and a ring-hydroxylated metabolite of parent. There was no free parent compound or phenol in non-acid-hydrolysed urine. In acid-hydrolysed urine, 61% of the dose was identified as phenol and 13% as 1-phenoxy-2-propanol. Although the parent compound was stable to acid hydrolysis, some of the phenol in acid hydrolysed urine may have arisen from degradation of acid-labile metabolite(s) as well as hydrolysis of phenol conjugates. 4. Rapid oral absorption, metabolism and urinary excretion of 1-phenoxy-2-propanol in rats were similar to other propylene glycol ethers. PMID- 14555342 TI - Decreased c-Myc expression and its involvement in X-ray-induced apoptotic cell death of human T-cell leukaemia cell line MOLT-4. AB - PURPOSE: To investigate the possible involvement of c-Myc and ceramide-c-Jun N terminal kinase (JNK) pathway in X-ray-induced apoptotic cell death of MOLT-4 cells. MATERIALS AND METHODS: The expressions of c-Myc protein and c-myc mRNA after X-irradiation were analysed by Western blotting and RT-PCR between radiosensitive MOLT-4 and radioresistant variant Rh-1a cells with less JNK activation than the parental cells. Apoptotic cell death was determined by a dye exclusion test, the appearance of chromatin condensation and DNA fragmentation. The effect of a JNK activator anisomycin or c-Myc inhibitor peptides (Int-H1-S6A, F8A) on the amount of c-Myc protein and on the induction of apoptosis was investigated, respectively. RESULTS: In X-irradiated MOLT-4 cells, amounts of both c-myc mRNA and c-Myc protein rapidly decreased, which was followed by apoptotic cell death, while little change or limited reduction of c-Myc protein was observed in X-irradiated Rh-1a cells with accompanying higher cell viability. Exposure of MOLT-4 and Rh-1a cells to c-Myc inhibitor peptides similarly induced apoptotic cell death with decreases of c-Myc protein. Anisomycin rapidly induced JNK activation and a subsequent decrease of c-Myc protein, causing cell death in MOLT-4 cells. On the other hand, Rh-1a cells were more resistant to anisomycin than parental MOLT-4 cells, showing less JNK activation and a delayed decrease of c-Myc protein. CONCLUSION: A decrease of c-Myc protein was considered important in X-ray-induced apoptotic cell death of MOLT-4 cells; activation of the JNK pathway caused reduction in the amounts of c-myc mRNA and c-Myc protein, and finally induced apoptotic cell death. PMID- 14555343 TI - Lovastatin causes sensitization of HeLa cells to ionizing radiation-induced apoptosis by the abrogation of G2 blockage. AB - PURPOSE: To investigate the effect of inhibition of Ras/Rho-regulated signalling by 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) on radiation-induced cell killing and apoptosis. MATERIALS AND METHODS: Different human cell lines were pretreated or not with lovastatin before exposure to gamma rays. Afterwards, radiation-induced cell killing, formation and repair of double strand breaks, activation of radiation-inducible signal mechanisms (i.e. p53, p21, extracellular-signal-related kinase (ERK), NF-kappaB), changes in cell cycle progression and apoptosis were analysed. RESULTS: As shown by a colony formation assay, lovastatin sensitized HeLa cells to gamma-radiation-induced cell killing. The lovastatin effect was cell-type specific. Neither the level of gamma-ray induced double-strand breaks nor its repair were affected by lovastatin. Sensitization was independent of p53/p21Waf1- and NF-kappaB-related mechanisms. Radiation-stimulated activation of ERKs was attenuated by lovastatin. Cell cycle analyses revealed that the level of gamma-ray-induced G2 blockage was not affected by lovastatin. However, as analysed up to 72 h after irradiation, lovastatin pretreated cells showed an accelerated abrogation of G2 blockage as compared with the control. G2 abrogation is paralleled by an increase in the frequency of apoptotic and necrotic cells. CONCLUSIONS: The data show that lovastatin can render human cells more sensitive to the cytotoxic effect of gamma rays. This is related to abrogation of G2 blockage and a concomitant increase in apoptotic/necrotic cell death. PMID- 14555345 TI - Comparison of the abundance of 10 radiation-induced proteins with their differential gene expression in L929 cells. AB - PURPOSE: To determine whether radiation-induced changes in protein abundance can be correlated with their differential gene expression in a murine fibroblast L929 cell line. MATERIALS AND METHODS: L929 cells were irradiated with 6 Gy. Cell lysates were collected at different points in time (20 min, 12, 24, 36, 48 and 72 h). The extracted proteins were separated by two-dimensional gel electrophoresis and quantified using computerized image analysis. Proteins exhibiting a differential expression equal to or more than twofold were identified by mass spectrometry following trypsin digestion. From these, 10 proteins characterized by large changes of radiation-induced abundance were selected in order to measure their corresponding gene expression using RTQ-PCR (real-time quantitative polymerase chain reaction). RESULTS: Up to 15-fold changes in the abundance of these 10 proteins were associated with no detectable changes more than twofold on the gene expression level. However, one gene (VEGF-D) showed a significant (p=0.005) up-regulation (1.8-fold). CONCLUSIONS: Deducing protein abundance from mRNA expression levels and vice versa appears to be of limited use. Furthermore, examination of transcriptional and translational changes provides different but complementary information. PMID- 14555344 TI - Low and high LET radiation-induced apoptosis in M059J and M059K cells. AB - PURPOSE: To investigate and compare the ability of DNA-dependent protein kinase (DNA-PK)-deficient and -proficient cells to undergo apoptosis after exposure to low and high linear energy transfer (LET) radiation. MATERIALS AND METHODS: A human glioma cell line M059J lacking the catalytic subunit of DNA-PK (DNA-PKcs) and its DNA-PKcs-proficient counterpart, M059K, were exposed to 1 and 4 Gy of accelerated nitrogen ions (14N, 140 eV nm(-1), 8-12 Gy min(-1)) or 60Co gamma rays (0.2 eV nm(-1), 0.7 Gy min(-1)). The induction of apoptosis was studied up to 144 h post-irradiation using two different methods: morphological characterization of apoptotic cells after fluorescent staining and cell size distribution analysis to detect apoptotic bodies. In parallel, protein expression of DNA-PKcs and poly(ADP-ribose) polymerase (PARP) as well as DNA-PK and caspase 3 activity were investigated. RESULTS: Low and high LET radiations (4 Gy) induced a time-dependent apoptotic response in both cell lines. Low LET radiation induced a significantly elevated apoptotic response in M059J as compared with M059K cells at 144 h post-irradiation. Following high LET radiation exposure, there was no difference between the cell lines at this time. PARP cleavage was detected in M059J cells following both low and high LET irradiation, while only high LET radiation induced PARP cleavage in M059K cells. These cleavages occurred in the absence of caspase-3 activation. CONCLUSIONS: M059J and M059K cells both display radiation-induced apoptosis, which occur independently of caspase-3 activation. The apoptotic course differs between the two cell lines and is dependent on the quality of radiation. PMID- 14555346 TI - Molecular nature of mutations induced by irradiation with repeated low doses of X rays in spleen, liver, brain and testis of lacZ-transgenic mice. AB - PURPOSE: To investigate the molecular characteristics of mutations induced by repeated low doses of X-rays in spleen, liver, brain and testis of mice. MATERIALS AND METHODS: Muta mice, which harbour the lacZ gene contained in the lambda genome, were irradiated with 0.15 Gy every Monday, Wednesday and Friday for 6 months starting at 10 weeks of age for a total of 78 times. Four months after the last irradiation, DNAs were isolated from the four different tissues and the mutant frequencies of lacZ were determined. Next, the nucleotide sequences of the mutant lacZ genes were determined and compared with that of the wild-type to identify the molecular changes in the mutants. The frequencies of different types of mutations were compared with those found in age-matched non irradiated mice. They were also compared with those found in mice irradiated with a single high dose. RESULTS: The repeated low-dose irradiation resulted in slight increases in the mutant frequency in the four kinds of tissues. The spleen, liver and brain in repeatedly irradiated mice showed higher frequencies of deletion type mutations than those of non-irradiated mice. In testis, however, the level of the increase was modest and not statistically significant. Complex type mutations were observed only in irradiated tissues. The characteristics observed in somatic tissues were similar to those induced by a single high dose of irradiation. CONCLUSIONS: The results suggest that the mechanism of mutation induction in vivo is similar between low- and high-dose irradiation in spleen, liver and brain. The low induction of deletion mutations in testis with low-dose irradiation suggests that spermatogonial cells have a unique DNA repair system against low-dose radiation-induced damage. PMID- 14555347 TI - Modelling study on the protective role of OH radical scavengers and DNA higher order structures in induction of single- and double-strand break by gamma radiation. AB - PURPOSE: To quantify the protective effects of (non-histonic) OH-radical scavengers and DNA higher-order structures in induction of single- (ssbs) and double-strand breaks (dsbs) by gamma-rays. MATERIALS AND METHODS: Spatial distributions of energy depositions by gamma-rays in liquid water were modelled with the track structure modules of the biophysical simulation code PARTRAC. Such distributions were superimposed on different DNA structure models (e.g. linear DNA, SV40 'minichromosomes' and compact chromatin), and direct energy depositions in the sugar-phosphate were considered as potential (direct) ssbs. The diffusion and interaction of the main chemical species produced in liquid water radiolysis were explicitly simulated, and reactions of *OH with the sugar-phosphate were considered as potential (indirect) ssbs. Two ssb on opposite DNA strands within 10 base pairs were considered as one dsb. Yields of ssb and dsb Gy(-1) Dalton(-1) in different DNA target structures were calculated as a function of the *OH mean lifetime, whose inverse value was taken as representative of the scavenging capacity of the DNA environment. RESULTS AND CONCLUSIONS: A further validation of the models implemented in the PARTRAC code has been provided, thus allowing a better understanding of the mechanisms underlying DNA damage. More specifically, the protection due to *OH scavengers was separately quantified with respect to that due to histones and chromatin folding, which could be 'switched off' in the simulations. As expected, for a given value of the environment scavenging capacity, linear DNA was more susceptible to strand breakage than SV40 minichromosomes, which in turn showed higher damage yields with respect to cellular DNA due to the larger accessibility offered to *OH. Furthermore, by increasing the scavenging capacity, the break yields decreased in all structures and tended to coincide with direct damage yields. Very good agreement was found with available experimental data. Comparisons with data on 'nucleoid' DNA (i.e. unfolded and histone-depleted DNA) also suggested that the experimental procedures used to obtain such structures might lower the environment scavenging capacity owing to the loss of cellular scavengers. PMID- 14555348 TI - Comparison of bleomycin and radiation in the G2 assay of chromatid breaks. AB - PURPOSE: To compare bleomycin with radiation in the G2 chromatid break assay. Controversy exists in the literature about whether G2 bleomycin chromatid-break sensitivity links with cancer predisposition in the same way as the G2 chromatid radiosensitivity test (the so-called 'G2 assay'). Although bleomycin is referred to as a 'radiomimetic' agent, it differs from radiation in the way the damage is induced. MATERIALS AND METHODS: Epstein-Barr virus-immortalized lymphoblastoid cell lines from two head and neck squamous cell carcinoma patients, two breast cancer patients, two ataxia-telangiectasia patients and two normal control persons were used. Chromosomal damage was determined in cells exposed to 0.3-Gy radiation or 5 mU ml(-1) bleomycin. The numbers of chromatid breaks per cell and of aberrations per cell (i.e. breaks and gaps) were determined. RESULTS: A strong positive correlation was found between the two different damage inducers (r=0.99; p<0.001). This correlation was similar for both the breaks per cell and the total aberrations per cell. Inclusion of gaps in the scoring of chromatid breaks was associated with a higher variability of the data, but this did not influence the outcome of this study. CONCLUSIONS: Both bleomycin and radiation give the same sensitivity phenotypes as determined by the G2 assay of chromatid breaks. Thus, when no radiation facility is present, bleomycin seems to be a good alternative to radiation for this type of assay. PMID- 14555349 TI - Nucleotide excision repair proteins and their importance for radiation-enhanced transfection. AB - PURPOSE: Irradiated cells transfect more efficiently than unirradiated cells because of a radiation-induced increase in plasmid integration. However, the molecular mechanism is unclear. Because of recent observations that nucleotide excision repair (NER) proteins can be involved in certain types of recombination in yeast, it was hypothesized that NER proteins might play a role in this radiation-enhanced integration. MATERIALS AND METHODS: Hamster and human cells with inactivating mutations in NER genes were irradiated at doses from 0 to 6 Gy and then immediately transfected with a linearized selectable marker plasmid. Transfection-enhancement ratios (TERs) were calculated as the ratio of the number of drug-resistant colonies in unirradiated cells to the number of transfectants in irradiated cells, corrected for cytotoxicity from radiation. RESULTS: Transfection into unirradiated rodent cells was unaffected by NER mutation status. Transfection into unirradiated human cells, however, was increased by NER mutation. The TERs were 5 and 100 for CHO and primary human fibroblasts, respectively, after exposure of the cells to 6 Gy. Mutations in ERCC1, XPA, XPB, XPC, XPF, XPG and CSB dramatically reduced TER. Mutations in ERCC1, XPC, XPF, XPG and CSB suppressed transfection so that the TER was significantly below 1. CONCLUSIONS: The mechanism of radiation-enhanced plasmid integration was distinct from that of plasmid integration in unirradiated cells, and NER gene products were critical for enhanced integration to occur. PMID- 14555350 TI - Difference in the heat sensitivity of DNA-dependent protein kinase activity among mouse, hamster and human cells. AB - PURPOSE: To examine the heat sensitivity of DNA-dependent protein kinase (DNA-PK) activity in a variety of cultured mouse, hamster and human cell lines. MATERIALS AND METHODS: Eight cell lines, which have been routinely used in our laboratory, were examined. Cells were heated at 44.0 +/- 0.05 degrees C and DNA-PK activity was measured by a DNA-pull-down assay followed by gel-electrophoresis. Cellular sensitivity to hyperthermia and/or X-ray was evaluated by a colony formation assay. RESULTS: In mouse FSA1233 and FM3A cells, DNA-PK activity dropped to 15 16% of unheated control after 20 min of heating. In Chinese hamster V79 and CHO K1 cells, kinase activity did not change appreciably after 20 min treatment but decreased to 60-70 and 22-23% after 40 or 60 min treatment, respectively. However, even after 180 min treatment, DNA-PK activity remained almost intact in human MOLT-4, MKN45 and A7 cells, and decreased only slightly in U937 cells. Hyperthermic radiosensitization was seen even in human cells but, as a trend, it was small compared with rodent cells. CONCLUSIONS: The heat sensitivity of DNA-PK was clearly different among mouse, hamster and human cells. The results suggested a possibility that the role of DNA-PK inactivation in hyperthermic radiosensitization might be variable, depending on cells, and would reinforce the warning that the direct extrapolation of data from rodent cells might lead to overestimation of the effectiveness of hyperthermia on human cancer. PMID- 14555351 TI - LET dependence of lethality of carbon ion irradiation to single tobacco cells. AB - PURPOSE: To determine the radiation sensitivity and relationship between linear energy transfer (LET) and relative biological effectiveness (RBE) in single plant cells irradiated with heavy ions. MATERIALS AND METHODS: Single cells were isolated from the tobacco BY-2 cell line and irradiated with carbon ions (78.6 309 keV microm(-1)) and gamma-rays (0.2 keV microm(-1)). Two weeks after irradiation, colonies with 16 cells or more derived from the irradiated cells were counted as survivors. The surviving fraction was fitted using the single hit, multitarget theory. RESULTS: The doses needed to reduce the surviving fraction of the cells to 0.1 (D10) of gamma-rays and carbon ions were 47.2 and 10.5-12.6 Gy, respectively. The RBE based on the D10 peaked at an LET of 247 keV microm(-1). The inactivation cross-section of carbon ions reached a plateau of 11.3 microm2 at an LET of 247 keV microm(-1). CONCLUSIONS: The radiation sensitivity of single tobacco cells was much lower than that of mammalian cells, although the mean number of base pairs per chromosome in the two cell types was similar. The RBE peak based on the D10 of carbon ions in single tobacco cells occurred at a higher LET than it does in other organisms. PMID- 14555353 TI - Introduction to the Monte Carlo project and the approach to the validation of probabilistic models of dietary exposure to selected food chemicals. AB - The Monte Carlo project was established to allow an international collaborative effort to define conceptual models for food chemical and nutrient exposure, to define and validate the software code to govern these models, to provide new or reconstructed databases for validation studies, and to use the new software code to complete validation modelling. Models were considered valid when they provided exposure estimates (e(a)) that could be shown not to underestimate the true exposure (e(b)), but at the same time are more realistic than the currently used conservative estimates (e(c)). Thus, validation required e(b) 15000 inhabitants) but there were fewer smokers among men in the smallest communities. The time trends indicated an overall decreasing serum cholesterol level and increasing body mass index for both sexes and a decreasing proportion of smokers among men during the years surveyed. CONCLUSIONS: There are differences in levels of cardiovascular risk factors between communities of different sizes in the Northern Sweden MONICA area, mainly to the disadvantage of the smallest communities. PMID- 14555371 TI - Smoking and associated factors among adolescents in Tallinn, Helsinki and Moscow: a multilevel analysis. AB - AIMS: To investigate variations in smoking and associated factors on individual and aggregate level among adolescents in Tallinn (Estonia), Helsinki (Finland) and Moscow (the Russian Federation). METHODS: The data was collected in Tallinn (N=1268 in Estonian and N=901 in Russian schools), in Helsinki (N=1396) and in Moscow (N=618) from pupils aged 13-18 by self-administered questionnaire. Multilevel analysis was used to estimate the possible effects of background characteristics, of the proximity of other smokers and the school context on the tobacco use of pupils. RESULTS: The prevalence rate of smoking among girls in Russian schools in Tallinn (34.6%), among girls in Helsinki (39.5%), and in Moscow (32.1%) was higher than that among girls in Estonian schools in Tallinn (17.6%). Smoking was slightly less prevalent among boys in Helsinki (27.5%), compared to boys in Estonian (33.6%) and Russian (35.6%) schools in Tallinn, and to boys in Moscow (32.8%). Smoking increased with age. Multilevel analysis showed that smoking differed by school among pupils. Individual determinants of smoking as study site, grade, friends', siblings' and parental smoking behaved differently depending upon school. Friends and siblings' smoking showed interaction with study site to the smoking among girls. CONCLUSIONS: Strategies aimed at influencing smoking behaviour need to be directed not only towards the individual, but also towards the influences within the child's school environment. PMID- 14555373 TI - High prevalence of tiredness and pain in young schoolchildren. AB - AIMS: This study presents the prevalence of tiredness, headache, stomach ache, and backache in young schoolchildren. METHODS: A randomized cluster sample of 1155 children from grade 0 to 6 (aged 6-13 years) in Umea, a city in northern Sweden, completed a questionnaire. RESULTS: Tiredness was the most frequently reported complaint. One-third (31%) of the children felt tired at least once a week, when starting school in the morning, and 16% felt tired almost every day. Headache was the most commonly reported pain: 23% reported weekly recurrent headache and 9% had headache more than once a week. However, in the two lowest grades, stomach ache was more common than headache. The prevalence of weekly recurrent stomach ache was 19% while 8% experienced stomach ache more than once a week. Recurrent monthly backache was reported by 18% of the children, approximately half of them reporting weekly complaints. In general, the prevalence of complaints increased by grade. Headache and stomach ache were more prevalent among girls compared with boys, while no gender difference was seen for tiredness and backache. CONCLUSIONS: This study shows that tiredness, headache, stomach ache, and backache are common also among young schoolchildren. The results suggest a need of preventive efforts addressing these complaints at early age. PMID- 14555374 TI - Work characteristics and morbidity as predictors of self-perceived health status in Norwegian physicians. AB - AIMS: The purpose of this study was to assess whether work characteristics and morbidity are predictors of physical, psychological, and social functioning in physicians. METHODS: A cross-sectional study was undertaken of 1126 Norwegian physicians. For comparison of health status scores, the Short Form 36 questionnaire was used, adjusting for differences in age and gender where applicable. Somatic morbidity was classified on the basis of self-reported conditions and psychiatric morbidity with score on the 25-item Hopkins symptom checklist using a cut-off indicating clinical psychiatric disease. Multiple logistic regression analysis was used with SF-36 dimension score below/above the 10th percentile as the dependent variable. RESULTS: In the multivariate model, psychiatric comorbidity was a significant predictor in 7 of 8 dimensions of health status (odds ratio 0.17 to 0.49), and somatic comorbidity in 6 of 8 dimensions of health status (odds ratio 0.33 to 0.59). Working hours was a significant predictor of only one dimension of health status, general health, after adjustment for demographic and work-related variables. CONCLUSIONS: The most important predictors for most dimensions of health status among Norwegian physicians were the presence or absence of self-reported somatic or psychiatric morbidity, while working hours, choice of specialty, and position were less important. PMID- 14555375 TI - Impaired fasting glucose and impaired glucose tolerance are related to both heredity and low birth weight. AB - AIM: The aim of the study was to test the hypothesis of an association between low birth weight (LBW), abnormal glucose metabolism and heredity. METHODS: Common subjects were identified as newborns in a local birth register and as adult participants in the Vasterbotten Intervention Programme (n=7796). Outcome measures such as impaired fasting glucose (IFG) and impaired glucose tolerance (IGT) were investigated in relation to LBW, family history of diabetes, and sex. RESULTS: Fasting plasma glucose and two-hour plasma glucose were elevated among women with LBW in relation to normal birth weight (NBW). Men and women with NBW had increased risks of IFG and IGT if there was a hereditary background of diabetes mellitus. LBW and hereditary background of diabetes mellitus interacted on the risk of IFG and IGT in women. CONCLUSIONS: Hereditary background of diabetes and LBW increase the risk of abnormal glucose metabolism such as IFG and IGT in early middle age. LBW and hereditary background of diabetes have a synergy effect on the risk of IFG and IGT in women. PMID- 14555376 TI - Developments in Scandinavian and American medical sociology. AB - This paper examines the similarities and differences in Scandinavian and American medical sociology. First, the issue of medicalization has not been as important in Scandinavian as it has been in American medical sociology. Second, women's health has been less explored in Scandinavian than in American medical sociology. Third, research on social inequalities and health has been a major focus in Scandinavian medical sociology since the 1990s. Fourth, cultural sociology and the poststructuralist perspective have been part of mainstream theory building in European sociology and also European and Scandinavian medical sociology during the past decade, while American medical sociology has been characterized by social-role theory and a quantitative approach. PMID- 14555377 TI - Are the "myths" of low back pain alive in the general Norwegian population? AB - AIM: The aim of this study was to examine the perception of low back pain care and consequences according to what Deyo refers to as seven "myths" about back pain, in the Norwegian population. METHODS: In spring 2001, seven questions, corresponding to Deyo's myths, were included in an opinion poll (telephone interviews) of a representative sample (n=1015) of the Norwegian population. RESULTS: In total, 41% of the population held that 'If you have a slipped disc you must have surgery'. Approximately 50% believed that 'X-ray and newer imaging tests can always identify the cause of pain' and 'Most back pain is caused by injury and heavy lifting'. Almost 60% agreed that 'Everyone with back pain should have a spine X-ray'. However, only one-quarter believed that 'If your back hurts, you should take it easy until the pain goes away', and approximately one-fifth believed that 'Back pain is usually disabling'. Only 12% believed that 'Bed rest is the mainstay of therapy'. More individuals in the lower- compared with the higher-educated groups believed in the myths. CONCLUSION: Information concerning current knowledge on healthcare and health consequences of low back pain had reached only a small part of the general population. The most important factor for lack of knowledge was education. Developing effective methods to promote adequate self-care and treatment and reduce the risk of chronicity of low back pain in the lower-educated groups should be a top priority. PMID- 14555379 TI - Tracking the colleges: the Australian Medical Council. PMID- 14555380 TI - The pathology of oral cancer. AB - Oral cancer is a serious public health problem, with over 200,000 new cases reported annually worldwide, two-thirds of which occur in developing countries. The overall mortality rate for intra-oral cancer remains high at approximately 50%, even with modern medical services, probably due to the advanced stage of the disease at presentation. This review outlines recent advances in our understanding of the roles and interactions of major risk factors for oral cancer worldwide, notably tobacco, alcohol and betel quid and the genetic polymorphisms determining their metabolism that may predispose patients to oral carcinoma. Oral epithelial lesions with malignant potential are described. We discuss the histopathology of oral cancer, its grading and the staging of the disease at clinical, microscopic, immunohistological and molecular levels. A recommended minimum dataset for pathology reports is described. The available strategies and current prospects for controlling oral cancer in the community are summarised. PMID- 14555381 TI - Differential expression of transforming growth factors-beta 1, -beta 2, -beta 3 and the type I, II, III receptors in the lining epithelia of inflamed gingiva. AB - AIMS: To investigate the distribution of transforming growth factor-beta isoforms in chronically inflamed periodontal tissues. METHODS: The present study determined, by immunohistochemistry, the expression patterns of TGF-betas and their receptors in the lining epithelia of inflamed gingiva. Frozen sections were obtained from 22 human gingival biopsies. RESULTS: TGF-beta 1 was not detected in gingival epithelial cells in examined sections. Detection of TGF-beta 2 indicated a progressive reduction of staining from the external oral epithelium through to gingival sulcus and the gingival attachment or pocket epithelium. TGF-beta 3 showed intense staining in all domains of both minimally inflamed gingiva and advanced periodontitis tissues. TGF-beta RI was visualised as focal staining of the spinous layer in the external oral epithelium of both periodontitis lesions and minimally inflamed tissues. TGF-beta RII was present throughout the strata, but with progressive reduction in intensity from the oral epithelium to gingival attachment or pocket epithelium respectively while, conversely, TGF-beta RIII showed an increase in diffuse staining intensity from external oral epithelium to pocket epithelium. CONCLUSIONS: A distinct expression profile was observed within different individuals for TGF-betas and the corresponding receptors. These findings provide a basis for evaluation of the role of these growth factors in the pathogenesis of periodontitis. PMID- 14555382 TI - Oral focal mucinosis: report of 15 cases and review of the literature. AB - AIMS: To describe 15 cases of oral focal mucinosis (OFM) and compare these to previously reported cases. METHODS: Cases diagnosed as OFM in the period 1981 2003-were reviewed. Clinical information provided at the time of submission of each specimen was retrieved and supplemented by additional clinical details provided by the respective clinician at the time of compilation of this paper. The literature was reviewed. RESULTS: OFM presented as an innocuous soft tissue swelling that may be either pedunculated or sessile. The gingiva was confirmed as the most common site for OFM, with a predominance of females affected. Microscopically, OFM is characterised by an area of myxoid tissue which is usually well-defined. The lesion is periodic acid-Schiff (PAS)-negative and alcian blue-positive, with pre-digestion with hyaluronidase preventing the alcian blue staining. As the differential diagnosis includes myxoid neural lesions, S100 staining is important in establishing the diagnosis, with cases of OFM being negative. CONCLUSIONS: The cause of OFM remains unknown. The cases presented in this paper bring OFM to the attention of anatomical pathologists when considering the differential diagnosis of myxoid lesions of the oral cavity. PMID- 14555383 TI - The value of HPV DNA typing in the distinction between adenocarcinoma of endocervical and endometrial origin. AB - AIMS: Distinguishing between adenocarcinomas of endocervical and endometrial origin histologically can be difficult, particularly in small biopsies. Most endocervical adenocarcinomas contain human papillomavirus (HPV) deoxyribonucleic acid (DNA) of 'high-risk' (HR) types, whereas this has not been consistently demonstrated in endometrial adenocarcinomas. The aim of this study was to determine whether HPV DNA testing could aid in this differential diagnosis. METHODS: The frequency of HPV DNA in paraffin-embedded tissue samples from 50 endocervical and 50 endometrial adenocarcinomas was investigated using polymerase chain reaction (PCR) amplification techniques involving (i) a screening HPV test followed by HPV DNA sequencing, and (ii) a test designed to detect HR genotypes 16, 18, 31, 33, 35, 45 and 58. Control specimens included cervical intraepithelial neoplasia (CIN) III lesions, squamous cell carcinomas (SCCs) of the cervix and lung, and colonic adenocarcinomas. Measures to minimise cross contamination were implemented. RESULTS: The screening test followed by HPV DNA sequencing had the highest sensitivity. By this test HR HPV DNA was detected in 11 of 11 (100%) cervical intraepithelial neoplasia (CIN III) lesions, nine of 10 (90%) cervical SCCs, none of 10 (0%) colorectal adenocarcinomas and none of 10 (0%) SCCs of the lung. Thirty-nine (78%) endocervical adenocarcinomas contained HR HPV DNA, compared to one (2.0%) endometrial adenocarcinoma. CONCLUSIONS: The results suggest that HPV DNA testing could be a useful adjunct in distinguishing between endocervical and endometrial adenocarcinomas in curettings or small biopsy specimens. PMID- 14555384 TI - Supporting the implementation of breast pathology recommendations: feasibility of a feedback mechanism to pathologists. AB - AIMS: The aims of the study were to explore the feasibility, acceptability, impact and cost of a feedback program to pathologists reporting on breast specimens through a BreastScreen Service. METHODS: The study was conducted at a single BreastScreen NSW Screening and Assessment Service. Pathology reports were audited against the ACN recommendations about pathology reporting [Australian Cancer Network Pathology Working Party. The Pathology Reporting of Breast Cancer: a guide for pathologists, surgeons and radiologists. Sydney: Australian Cancer Network, 1997] during two consecutive periods. Feedback was provided to pathologists at the completion of each audit. A comparison of completeness of pathology reports between audits was undertaken. The Screening and Assessment Service staff and participating pathologists were interviewed to assess acceptability and information about resource allocation. RESULTS: The study demonstrated that the feedback mechanism was acceptable to pathologists and staff at the BreastScreen Screening and Assessment Service, and that participation in the study resulted in some improvements in pathology reporting. CONCLUSIONS: The study demonstrated the feasibility of providing feedback to pathologists about their reporting of breast specimens. However, there were costs to the Service of participating. A way of managing these costs would need to be explored if the feedback mechanism was made a routine BreastScreen procedure. PMID- 14555385 TI - The myopathology of floppy and hypotonic infants in Singapore. AB - AIMS: This study attempts to determine the type and relative frequency of muscle diseases contributing to floppy and hypotonic infants in Singapore. METHODS: Eighty consecutive muscle biopsies in the Department of Pathology, National University of Singapore, in the period 1978-2000, in which a clinical diagnosis of floppy or hypotonic infant was made, were reviewed. RESULTS: The commonest cause of severe hypotonia in infancy was spinal muscular atrophy, which accounted for 33% of cases followed by congenital muscular dystrophy (13%). Eight cases (10%) of infantile type II glycogenosis (Pompe's disease) were encountered. There were seven cases of congenital myopathy, of which four were centronuclear myopathy, and one each of central core myopathy, nemaline myopathy and congenital fibre type disproportion. One case of centronuclear myopathy was associated with type I fibre smallness. Type II atrophy, which is generally considered a non specific change, was encountered in five cases. Of interest is the relatively large number of muscle biopsies (29%) in which no significant pathological features were encountered at the light microscopic, histochemical as well as ultra-structural level. CONCLUSIONS: The study has revealed a great variety of pathology affecting the muscle of children presenting as floppy infants or with hypotonia. The muscle diseases included spinal muscular atrophy, congenital muscular dystrophies, congenital myopathies and metabolic myopathies. However, 23 (29%) cases showed no significant pathology. For this group of floppy and hypotonic infants further studies are needed. PMID- 14555386 TI - Improvement in the detection rate of t(14;18) translocation on paraffin-embedded tissue: a combination approach using PCR and FISH. AB - AIMS: PCR has been the primary method used for the detection of t(14;18) translocation in formalin-fixed, paraffin-embedded tissues. This technique mainly targets the well-characterised breakpoint regions in chromosomes 14 and 18. FISH is now applicable on paraffin tissue sections and has been suggested to be capable of detecting essentially 100% of t(14;18) translocated cases. In this study, we described the application of both PCR and FISH for the detection of t(14;18) translocation. METHODS: Fifty follicular lymphoma cases were retrieved from the files of the Department of Pathology, University of Malaya Medical Centre (UMMC). Nested PCR amplification of MBR/JH and mcr/JH was performed in these cases, and those cases that did not demonstrate the translocation were subjected to FISH analysis. RESULTS: Thirty cases (60%) had t(14;18) translocation detected by PCR, 25 (50%) had breakpoint with MBR and five (10%) involved mcr. Twenty cases without detectable t(14;18) translocation by PCR were analysed by FISH. Eleven cases were successfully probed, and four of them showed positive translocation signal. CONCLUSIONS: The combination of PCR and FISH analysis on paraffin tissue sections for the detection of t(14;18) translocation increases the sensitivity of detection from 60 to 68%. Problems encountered in our FISH analysis on tissue sections impose certain limitations in using this technique for retrospective screening of large number of samples. Therefore, we suggested the application of PCR as the first screening tool on retrospective archival materials, followed by FISH on those PCR-negative cases. PMID- 14555387 TI - Epithelial membrane antigen (EMA) or MUC1 expression in monocytes and monoblasts. AB - AIMS: Epithelial membrane antigen (EMA) or MUC1 belongs to a heterogeneous group of heavily glycosylated proteins and is expressed in most normal and epithelial neoplastic cells. EMA is also expressed in plasma cells, anaplastic large cell lymphoma (Ki-1 antigen), malignant histiocytosis and erythroleukaemia. In 1996, Cheong et al. (Hematology 1996; 1: 223) demonstrated the positive expression of EMA in monoblasts. Since there were very few useful markers for differentiating subtypes of acute myeloid leukaemia with a monocytic component from the those without, a study was conducted to evaluate the prevalence of EMA expression and its relationship with known markers for monocytic-macrophage lineage (CD11c, CD14 and intracellular CD68) in monocytes and monoblasts. METHODS: EMA detection was performed by flow cytometry in monocytes and monoblasts. EMA expression was compared with other known markers of monocytic-macrophage lineage (CD11c, CD14 and intracellular CD68). Samples of purified monocytes were obtained from 20 healthy volunteers. Twenty-two cases of monocytic AML (M4 and M5) were studied and controls were selected from 20 cases of acute lymphoblastic leukaemia (ALL) and 18 cases of non-monocytic AML (M0, M1, M2, M3, and M7). RESULTS: EMA was shown to be expressed strongly on the surface of all purified monocytes. EMA expression was observed on blast cells in 18/22 (81.8%) cases of AML M4 and M5, but not in that of non-monocytic AML or ALL. In this study EMA monoclonal antibody has demonstrated a strong association (P<0.001) with all the other known markers of monocytic-macrophage lineage in acute leukaemia subtypes. EMA had also shown 100% specificity and 81.8% sensitivity in the diagnosis of AML M4 and M5. CONCLUSIONS: The monoclonal antibody EMA (clone E29) is a useful marker in the classification of acute myeloid leukaemia and can be used as a supplementary analysis for the diagnosis of acute leukemia with monocytic involvement. PMID- 14555388 TI - The role of CD30, CD40 and CD95 in the regulation of proliferation and apoptosis in classical Hodgkin's lymphoma. AB - AIMS: CD30, CD40 and CD95 are members of the tumour necrosis factor receptor superfamily. Ligation to their respective ligands (CD30L, CD40L, CD95L) will generate a diverse set of signalling cascades. We aim to study the expression pattern of CD30, CD40 and CD95 in classical Hodgkin's lymphoma (cHL) and to correlate the expressions with proliferation and apoptosis in the Hodgkin/Reed Sternberg (H/RS) cells of cHL with or without associated Epstein-Barr virus (EBV) infection. METHODS: A total of 66 cHL cases were retrieved from the archives. Expressions of CD30, CD40, CD95 and proliferation by Ki-67 expression were detected with an immunohistochemical staining method. Apoptosis index was assessed by in situ TUNEL staining technique on 30 randomly selected cases and the presence of EBV was determined by EBER in situ hybridisation. RESULTS: Expression of CD30, CD40 and CD95 in the H/RS cells was observed in a high proportion of the cases (100, 93.9, 90.5%, respectively). There was no significant association or correlation of the expression of these molecules with the presence of EBV. Expression of CD40 was associated with expression of the proliferation marker Ki-67 (P=0.044), whereas strong (intermediate and high) expression of CD30 showed a significant correlation with proliferation in the EBV negative cases only (P=0.025). No correlation was observed for the expression of CD30 and CD40 with apoptosis of the H/RS cells. The childhood cases showed weaker CD95 expression in the H/RS cells than the adult cases, and the expression of CD95 was weaker than that of CD40 in the childhood group. CONCLUSIONS: Our results showed that CD30, CD40 and CD95 are highly expressed in the H/RS cells of the majority of cases of cHL. The expression patterns seem to be independent of EBV and do not correlate with apoptosis of the H/RS cells. PMID- 14555390 TI - Test and teach. An unusual cause of multiple bilateral pulmonary metastases. Osteosarcoma arising in monostotic Paget's disease of the femur. PMID- 14555389 TI - Common ALK gene rearrangement in Asian CD30+ anaplastic large cell lymphoma: an immunohistochemical and fluorescence in situ hybridisation (FISH) study on paraffin-embedded tissue. AB - AIMS: The most common recurrent genetic aberration in anaplastic large cell lymphoma (ALCL) is translocation involving the ALK gene that results in ectopic expression of ALK protein in lymphoid tissue. This study aims to investigate the frequency of ALK gene rearrangement in a series of Asian ALCL. METHODS: ALK gene rearrangement was detected by immunostaining of ALK protein and fluorescence in situ hybridisation (FISH) targeting at the 2p23 region. RESULTS: The expression of ALK protein was detected in 24/34 (71%) of the cases, and it was significantly higher in childhood cases (100%) when compared to adult cases (47%). The analyses by FISH were consistent with the results from immunostaining of ALK protein, but the analyses were only successful in 15/34 (44%) cases. FISH analyses detected extra copies of ALK gene in three cases, including one case that expressed ALK protein and showed 2p23 rearrangement. CONCLUSIONS: The current series revealed a high frequency of ALK gene rearrangement, especially in the children. Immunostaining of ALK protein is a reliable indication of ALK gene rearrangement, and is superior to FISH. However, FISH analysis is useful in detecting other genetic aberrations that are not related to ALK gene rearrangement. PMID- 14555392 TI - Immunohistochemical detection of the human herpes virus 8 (HHV8) latent nuclear antigen-1 in Kaposi's sarcoma. AB - AIMS: The molecular genetics of the human herpes virus 8 (HHV8) has now been characterised and the virus appears to be important in the pathogenesis of Kaposi's sarcoma (KS). This study attempts to determine the rate of HHV8 infection in KS in an Australian cohort. METHODS: Routine streptavidin-biotin peroxidase immunostaining with diaminobenzidine was performed on paraffin embedded archival tissues of 37 KS cases using a murine monoclonal antibody directed against the C-terminus of the latent nuclear antigen-1 molecule of HHV8 (clone 13B10; Novocastra) at 1:50 dilution. RESULTS: Positive HHV8 nuclear staining was detected in the nuclei of the spindle cells and endothelial cells of the vascular channels in about 78% (29/37) of all cases. HHV8 staining was absent in the non-neoplastic vessels in the adjacent tissue (P=0.0001, chi(2)=44.46; chi(2)-test with continuity correction) and the negative control cases of Merkel cell carcinoma (P=0.02, chi(2)=5.07; chi(2)-test with continuity correction). HHV8 staining was detected in 80% (8/10 cases) of the patch stage, 88% (7/8 cases) of the plaque stage and 74% (14/19 cases) of the late stage. No significant difference was found between HHV8 positivity and HIV status, age, gender, tumour recurrence, multiplicity or site of the lesions. CONCLUSIONS: The latent nuclear antigen-1 of HHV8 can be detected by immunohistochemistry in the majority of human KS lesions, raising the possibility of its future potential use as an adjunct for the diagnosis of KS in problematic cases. PMID- 14555393 TI - Primary meningeal anaplastic large cell lymphoma. PMID- 14555394 TI - Mycoplasma hominis infection of a subdural haematoma in the peripartum period. PMID- 14555396 TI - Re: Quantitative evaluation of AgNORs in bone tumours. PMID- 14555395 TI - Smooth muscle hyperplasia of the epididymis: a case report. PMID- 14555398 TI - Oral bioavailability of pentachlorophenol from soils of varying characteristics using a rat model. AB - Evidence accumulated during the last two decades suggests that only a fraction of any chemical in soil is available to organisms, and soil-related effects on bioavailability should be considered in optimizing soil remediation cleanup levels. In the current study, the absolute and relative bioavailabilities of pentachlorophenol (PCP) from freshly spiked and environmentally aged soils varying in organic carbon content, clay content, and pH were examined using a rat model. PCP is a broad-spectrum biocide widely used as a wood preservative, and thus is ubiquitous in the environment. Soils and corn oil containing equivalent levels of PCP were administered to male Sprague-Dawley rats by gavage at 2 dose levels: 100 and 200 microg/kg body weight. Equivalent doses were also given intravenously. The areas under the plasma concentration of PCP versus time curves were calculated, and absolute and relative bioavailabilities of PCP from each soil were determined. At a dose of 100 microg/kg body weight, the absolute bioavailabilities of PCP across soils ranged from 36% to 65%, and the relative bioavailabilities ranged from 48% to 82%. At the higher dose level (200 microg/kg body weight), absolute and relative bioavailability ranges were somewhat higher at 46% to 77% and 52% to 87%, respectively. All soils decreased absolute PCP bioavailability significantly at both dose levels and relative bioavailability at the lower PCP dose level. At the higher dose level, only one of the two field contaminated soils decreased the relative PCP bioavailability. The data indicate that PCP-soil interactions do significantly decrease the oral bioavailability of PCP from soil, but no obvious correlation was observed between soil properties and bioavailabilities. PMID- 14555399 TI - Health of tree swallow (Tachycineta bicolor) nestlings exposed to chlorinated hydrocarbons in the St. Lawrence River Basin. Part II. Basal and stress plasma corticosterone concentrations. AB - Sixteen-day-old tree swallows (Tachycineta bicolor) were collected in 1999 and 2000 from nine sites within the vicinity of the St. Lawrence River in Canada and the United States to determine if organochlorine contaminant concentrations correlated with corticosterone levels. Basal plasma corticosterone levels were determined in chicks reared in nest boxes, and stress plasma corticosterone levels were determined in chicks after exposure to a 10-min standardized stress test. Mean basal plasma corticosterone concentrations ranged from 4 to 37 ng/ml in 1999, and from 5 to 20 ng/ml in 2000. Mean poststress plasma corticosterone levels ranged from 26 to 67 ng/ml in 1999, and from 26 to 109 ng/ml in 2000. In 1999 and 2000, basal corticosterone was negatively correlated with polychlorinated dibenzofuran (PCDF). Total PCDFs ranged from 4.8 to 120.5 ng/kg wet weight in chicks in 1999 and 2000 among sites. These results indicate that current levels of organochlorine contaminants in the St. Lawrence River and surrounding tributaries may be interfering with the glucocorticoid endocrine axis of tree swallows. PMID- 14555400 TI - Metallothionein isoform 1 and 2 gene expression in a human urothelial cell line (UROtsa) exposed to CdCl2 and NaAsO2. AB - Studies have shown that metallothionein (MT) is overexpressed in some human bladder cancers and that overexpression can predict treatment response to neoadjuvant cisplatin, methotrexate, and vinblastine chemotherapy. In the present study the UROtsa cell line, a model of normal human urothelium, was used to determine the expression of the human MT-1 and MT-2 genes and MT protein following exposure to CdCl(2) or NaAsO(2) at lethal and sublethal levels. Acute exposure was modeled by treating confluent cultures with 100 microM NaAsO(2) or 53.4 microM CdCl(2) for 4 h followed by a 48-h recovery period. Extended exposure was modeled by treating confluent cells with 1, 4, and 8 microM As(3+) or 1, 5, and 9 microM Cd(2+) for 16 d, with the highest concentrations producing cell lethality. The expression of MT mRNAs and protein were determined by reverse transcription polymerase chain reaction (RT-PCR) and immunoblot analysis. Cell viability was determined by the MTT assay. It was shown that acute exposure to either As(3+) or Cd(2+) increased the levels of mRNAs for the MT-1E, MT-1X, and MT-2A genes, whereas extended exposure only increased these mRNAs following exposure to Cd(2+). It was shown that both acute and extended exposure to either As(3+) or Cd(2+) increased the levels of MT protein, reaching a maximal value of 8 ng MT protein/microg total protein for acute exposure to Cd(2+). This is in contrast to previous studies using cultured human proximal tubule cells, where similar extended treatment with Cd(2+) resulted in over 20-fold higher MT protein levels. These studies demonstrate that human urothelial cells accumulate only modest amounts of MT protein when exposed to either Cd(2+) or As(2+) for a 16-d time period. PMID- 14555401 TI - Increased expression of glial fibrillary acidic protein in cerebellum and hippocampus: differential effects on neonatal brain regional acetylcholinesterase following maternal exposure to combined chlorpyrifos and nicotine. AB - Cigarette smoking and environmental exposure to chlorpyrifos during pregnancy could lead to developmental toxicity in the offspring. In the present study, pregnant female Sprague-Dawley rats (300-350 g) were treated daily with nicotine (1 mg/kg, sc) or chlorpyrifos (0.1 mg/kg, dermal) or a combination of nicotine and chlorpyrifos from gestational days (GD) 4-20. Control animals were treated with saline and ethanol. Male offspring from the mothers treated with nicotine alone gained significantly less weight on postnatal day (PND) 30 as compared to control. On PND 7, there was a significant increase in brain acetylcholinesterase (AChE) activity in pups from nicotine- and chlorpyrifos-treated dams, whereas plasma butyrylcholinesterase (BChE) activity was significantly elevated in pups of mothers treated with either chlorpyrifos alone or pesticide combined with nicotine. On PND 30 there was a significant increase in AChE activity in brainstem and cerebellum in all treated male pups. In female pups on PND 30 there was a significant rise in AChE activity in brainstem of chlorpyrifos alone and in cerebellum of the combination nicotine and chlorpyrifos group. Histopathological evaluation demonstrated an increased neuronal cell death in the cerebellum granular cell layer of female offspring from nicotine or combined nicotine with chlorpyrifos group. A rise in glial fibrillary acidic protein (GFAP) immunostaining was observed in the CA1 subfield of hippocampus and cerebellum on PND 30 in female and male offspring of mothers treated with either nicotine or nicotine in combination with chlorpyrifos, but to a lesser extent in males. Data suggest that maternal exposure to nicotine and chlorpyrifos, alone or in combination, produces differential alterations in brain regional AChE activity and expression of GFAP in cerebellum and hippocampus in offspring on PND 30. PMID- 14555402 TI - Dioxin, dibenzofuran, and coplanar PCB levels in Laotian blood and milk from agent orange-sprayed and nonsprayed areas, 2001. AB - Agent Orange, a phenoxyherbicide contaminated with 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD), was used by American military forces during the United States Vietnam war between 1962 and 1971 primarily as a defoliant to destroy forests where enemy troops might find cover. Agent Orange was used mainly in Vietnam, but also to a lesser extent in Laos and Cambodia. In Laos, there have been no prior studies of TCDD contamination from Agent Orange, despite known defoliation and documented records of Agent Orange spraying. This article presents findings of TCDD in human blood and milk from two geographic areas in Laos: Vientiane, a nonsprayed area, and Sepone, an Agent Orange-sprayed area. German and Canadian laboratories used high-resolution gas chromatography-mass spectrometry to measure 7 dioxin, 10 dibenzofuran, and 4 non-ortho or coplanar polychlorinated biphenyls in Laotian blood and milk samples. Most subjects tested in this Laos Ministry of Health Study showed low dioxin and dibenzofuran levels, consistent with what would be expected in a primarily rural nonindustrial country. These findings are consistent with relatively low dioxin and dibenzofuran levels recently found in food from these same areas. The chemically and toxicologically related non-ortho PCBs were measured but were found at low levels compared to specimens from other countries, presumably because of less industrialization and industrial pollution in Laos. PMID- 14555403 TI - Effect of neonatal rat bisphenol a exposure on performance in the Morris water maze. AB - Bisphenol A (BPA), an environmental estrogen, is a component of many food and beverage containers and can leach into the container contents over time. Due to its estrogenic properties, exposure to BPA during development could alter the appropriate maturation of pathways essential for normal cognitive function at later ages. To investigate this, the effects of repeated postnatal exposure of male and female rats to BPA on spatial learning and memory were investigated using a Morris water maze. Breeders and offspring were maintained on a standard phytoestrogen-free diet. Oral administration of 72 microg/kg 17 beta-estradiol (E(2)), 100 microg/kg BPA (low BPA), 250 microg/kg BPA (high BPA), or the safflower oil vehicle was performed daily from postnatal d 1 (PND1) through PND14. There were no treatment-related effects on swimming ability or motivation (PND33) or on acquisition of maze solution (PND34-37). However, acquisition of maze performance was significantly better in control males than in control females. Treatment with E(2) and low BPA disrupted this normal gender-dependent pattern of acquisition, while treatment with high BPA did not. In a probe trial (PND40), females treated with high BPA spent significantly less time in the escape quadrant. These data indicate that E(2) and low dosages of BPA can alter the normal gender-dependent pattern of acquisition, while higher dosages of BPA alter the retention of spatial information without significantly affecting acquisition. PMID- 14555405 TI - Carboplatin-induced oxidative injury in rat inferior colliculus. AB - Carboplatin is currently being used as an anticancer drug against human cancers. However, high dose of carboplatin chemotherapy resulted in ototoxicity in cancer patients. Carboplatin-induced ototoxicity was related to oxidative stress to the cochlea and inner hair cell loss in animals. It is likely that initial oxidative injury spreads throughout the neuroaxis of the auditory system later. The study aim was to evaluate carboplatin-induced hearing loss and oxidative injury to the central auditory system (inferior colliculus) of the rat. Male Wistar rats were divided into two groups of seven animals each and treated as follows: (1) control (normal saline, intraperitoneal [i.p.]) and (2) carboplatin (256 mg/kg, i.p.). Auditory brain-evoked responses (ABRs) were recorded before and 4 days after treatments. The animals were sacrificed on the 4th day and inferior colliculus from brain stem and cerebellum were isolated and analyzed. Carboplatin significantly elevated the hearing threshold shifts at clicks, 2-, 4-, 8-, 16-, and 32-kHz tone burst stimuli. Carboplatin significantly increased nitric oxide and lipid peroxidation, xanthine oxidase, and manganese superoxide dismutase activities in the inferior colliculus, but not in the cerebellum, indicating an enhanced flux of free radicals in the central auditory system. Carboplatin significantly depressed the reduced to oxidized glutathione ratio, antioxidant enzyme activities, such as copper-zinc superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, and glutathione S-transferase, and enzyme protein expressions in the inferior colliculus, but not in the cerebellum, 4 days after treatment. The data suggest that carboplatin induced oxidative injury specifically in the inferior colliculus of the rat leading to hearing loss. PMID- 14555406 TI - Effect of ketamine on cocaine-induced immunotoxicity in rats. AB - The abuse of cocaine (COC) with ketamine (KET) is currently popular among young drug abusers and has been associated with increased risk of human immunodeficiency virus (HIV) infection. The effect of subacute exposure to COC and KET alone and in combination on the immune system was assessed in adult male Sprague-Dawley (SD) rats. To simulate the route and mode of human exposure, rats were treated with COC alone (5 mg/kg, i.v.), KET alone (100 mg/kg, p.o.) or KET followed immediately by COC (same doses and routes of administration) once-a-day for 7 consecutive days. Rats were sacrificed 30 minutes following the last treatment. Total circulating leukocyte and lymphocyte counts were decreased with relative neutrophilia, whereas immunoglobulin M (IgM) antibody response to sheep erythrocytes (SRBCs) was increased in animals treated with COC. Moreover, treatment with COC alone increased serum interleukin-10 (IL-10) concentration; however, it did not affect serum interferon gamma (INF-gamma) concentration. Spleen histology showed hyperplasia of white pulp whereas thymus gland demonstrated mild cortical degeneration. On the other hand, KET treatment did not produce any significant change of any of these parameters. However, when coadministered with COC, significant reduction of bodyweight, spleen/bodyweight, and thymus/bodyweight ratios with degeneration of splenic white pulp and thymic cortex occurred. Moreover, the primary immunoglobulin response to SRBC and serum IL-10 concentration were decreased without significant change in serum IFN-gamma or circulating leukocytic counts. COC caused a significant increase in serum corticosterone concentration that KET effectively prevented. On the other hand, a significant increase in plasma and tissue concentrations of norcocaine (NC) resulted following KET and COC administration in combination. Daily SKF-525A pretreatment at a dose of 30 mg/kg, i.p., for 7 days 1 hour prior to KET and COC in combination effectively reversed the effects of this combination on body weight, organ/bodyweight ratios, histopathology, and serum IgM and IL-10 concentrations without affecting leukocytic counts. On the other hand, SKF-525A pretreatment did not change the immunomodulatory effects of COC compared to non pretreated animals. The results suggest that COC-induced immunomodulation most likely occurred through neuroendocrinal mechanisms. On the other hand, enhanced oxidative metabolism of COC in the presence of KET-induced immunosuppression. PMID- 14555409 TI - The use of unallowed excipients. AB - The laws governing the U.S. Food and Drug Administration (FDA) do not provide for the approval of stand-alone excipients. Current regulations do not permit the use of excipients, but allow their use. The acceptance process for excipients is slow and only recently did the FDA propose draft guidance for nonclinical studies for pharmaceutical excipients. The FDA has made four suggestions to the U.S. Pharmacopoeia concerning including excipient monographs in the National Formulary for excipients not yet allowed. This article reviews these four proposals to identify the proposal that is most appropriate. PMID- 14555407 TI - Immunohistochemical changes in the mouse striatum induced by the pyrethroid insecticide permethrin. AB - Epidemiological studies have linked insecticide exposure and Parkinson's disease. In addition, some insecticides produce damage or physiological disruption within the dopaminergic nigrostriatal pathway of non-humans. This study employed immunohistochemical analysis in striatum of the C57BL/6 mouse to clarify tissue changes suggested by previous pharmacological studies of the pyrethroid insecticide permethrin. Dopamine transporter, tyrosine hydroxylase, and glial fibrillary acidic protein immunoreactivities were examined in caudate-putamen to distinguish changes in amount of dopamine transporter immunoreactive protein from degeneration or other damage to dopaminergic neuropil. Weight-matched pairs of pesticide-treated and vehicle-control mice were dosed and sacrificed on the same days. Permethrin at 0.8, 1.5 and 3.0 mg/kg were the low doses and at 200 mg/kg the high dose. Brains from matched pairs of mice were processed on the same slides using the avidin-biotin technique. Four fields were morphometrically located in each of the serial sections of caudate-putamen, digitally photographed, and immunopositive image pixels were counted and compared between members of matched pairs of permethrin-treated and vehicle-control mice. For low doses, only 3.0 mg/kg produced a significant decrease in dopamine transporter immunostaining. The high dose of permethrin did not produce a significant change in dopamine transporter or tyrosine hydroxylase immunostaining, but resulted in a significant increase in glial fibrillary acidic protein immunostaining. These data suggest that a low dose of permethrin can reduce the amount of dopamine transporter immunoreactive protein in the caudate-putamen. They also suggest that previously reported reductions in dopamine uptake of striatal synaptosomes of high-dose mice may be due to nondegenerative tissue damage within this region as opposed to reductions of dopamine transporter protein or death of nigrostriatal terminals. These data provide further evidence that insecticides can affect the primary neurodegenerative substrate of Parkinson's disease. PMID- 14555410 TI - Toxicity of excipients--a Food and Drug Administration perspective. AB - Excipients are essential components of drug products. They are also potential toxicants. Examples of known excipient-induced toxicities include renal failure and death from diethylene glycol, osmotic diarrhea caused by ingested mannitol, hypersensitivity reactions from lanolin, and cardiotoxicity induced by propylene glycol. Proposals to test or market new drug products in the United States should adequately address the safety of the proposed exposure to the excipients in those products. The specific safety data that may be needed will vary depending upon the clinical situation, including such factors as the duration, level, and route of exposure, but may include acute, repeat-dose, reproductive, and genetic toxicity data, carcinogenicity data, and specialized toxicology information, such as sensitization or local irritation data. Many guidances exist to aid in the development of pharmaceuticals, including the International Conference on Harmonization (ICH) documents and various Food and Drug Administration/Center for Drug Evaluation and Research (FDA/CDER) pharmacology and toxicology guidances. The FDA/CDER has recently adopted a new guidance for industry, "Nonclinical Studies for Development of Pharmaceutical Excipients," which focuses on issues associated with development of safety databases that will support clinical use of excipients in drug products. The new guidance document is introduced and discussed in this article. PMID- 14555411 TI - Are dietary supplements safe as currently regulated? The great debate. AB - At its 2002 annual meeting, the American College of Toxicology hosted in its series of issue sessions a symposium entitled "The Great Debate: Are Dietary Supplements Adequately Regulated to Enhance the Safety of Consumers." There were five participants in the debate, of whom three (the moderator and one advocate from either side) contributed papers. This article, reflecting the author's talk as the moderator, presents the applicable regulations, an overview, and history of the marketplaces and the range of perceived consumer safety problems. PMID- 14555412 TI - The Dietary Supplement and Health Education Act and supplements: dietary and nutritional supplements need no more regulations. AB - The Dietary Supplement and Health Education Act (DSHEA) of 1994 was intended to ensure consumer access to safe dietary supplements, e.g., herbs, minerals, and vitamins. It granted the Food and Drug Administration (FDA) considerable enforcement authority to regulate dietary supplement industry. This article discusses relevant components of the DSHEA, explains the division of regulatory responsibilities between the FDA and the Federal Trade Commission (FTC), and refutes the often cited allegation that the herb and dietary supplement industry are unregulated. PMID- 14555413 TI - Why consumers need more protection against claims for dietary supplements and herbs. AB - Federal agencies have never had enough resources to cope with the enormous amount of deception in the supplement and health-food marketplace. Passage of the 1994 Dietary Supplement Health and Education Act (DSHEA) has worsened the situation by encouraging dubious claims and weakening the Food and Drug Administration (FDA)'s ability to ban dangerous products. PMID- 14555414 TI - Parkinsonism due to manganism in a welder. AB - A 33-year-old right-handed male presented complaining of a 2-year history of progressive cognitive slowing, rigidity, tremors, slowing of movements, and gait instability leading to falls. On examination, he had a Mini-Mental Status Examination (MMSE) score of 29, slowed saccadic eye pursuit, hypomimia, cogwheel rigidity, a 3- to 4-Hz tremor, and a "cock-walk" gait. His symptoms and signs were similar to idiopathic Parkinson's disease; however, he was young, inattention and forgetfulness occurred early in the course of the disorder, levodopa was unhelpful, and his gait was atypical. His work up for secondary causes of parkinsonism was negative, except for increased signal intensity on T1 weighted magnetic resonance image (MRI) in the bilateral basal ganglia. Typical etiologies for that finding were ruled-out, which led to further inquiries into the patient's lifestyle. He was a welder, and discussion with his employer revealed that he used a steel-manganese alloy, he often worked in a confined ship's hold, and he did not use a respiratory mask. Because manganese toxicity can produce increased T1-weighted signal intensities in the basal ganglia, the authors tested his serum and urine manganese, and both were elevated. This patient emphasizes the importance of a careful occupational history in persons presenting with atypical manifestations of a neurodegenerative disorder. It also lends support to the hypothesis that welding can produce enough exposure to manganese to produce neurologic impairment. PMID- 14555415 TI - A toxicity profile of osteoprotegerin in the cynomolgus monkey. AB - Osteoprotegerin (OPG) is a novel secreted glycoprotein of the tumor necrosis factor (TNF) receptor superfamily that acts as an antiresorptive agent inhibiting osteoclast maturation. OPG acts by competitively inhibiting the association of the OPG ligand with the RANK receptor on osteoclasts and osteoclast precursors. This inhibition of osteoclasts can lead to excess accumulation of newly synthesized bone and cartilage in vivo. The purpose of this study was to investigate the potential toxicity of a human recombinant form of OPG in the young cynomolgus monkey. OPG was administered by intravenous (i.v.) or subcutaneous (s.c.) injection three times per week for either 4 or 13 weeks. There were no deaths during the study, no clinical signs related to treatment, no effect on body weight, appetence, or ophthalmology. No toxicologically relevant changes in routine laboratory investigations, organ weights, or gross or histopathological findings were observed. Serum ionized calcium and phosphorus were decreased at all dose levels. Evaluations were performed to monitor biochemical markers of bone resorption (N-telopeptide [NTx], deoxypyridinoline [DPD]), bone formation (skeletal alkaline phosphatase [sALP], osteocalcin [OC]), parathyroid hormone [PTH], and bone density of the proximal tibia and distal radius in vivo. Dose-related decreases in NTx and/or DPD were observed at each dose level, with up to a 90% decrease in NTx noted for animals treated i.v. or s.c. at 15 mg/kg. Similar decreases were observed for sALP and OC. PTH was increased for animals treated at 5 and 15 mg/kg (i.v. or s.c.). Trabecular bone density was increased for the majority of males and females treated i.v. or s.c. at 15 mg/kg and males treated i.v. at 5 mg/kg. Microscopic examination of the sternebrae revealed corresponding increases in bone. Decreases in markers of bone turnover, and corresponding increases in bone density, were consistent with the pharmacological action of OPG as an osteoclast inhibitor. The no-observable adverse-effect level (NOAEL) of OPG was 15 mg/kg. PMID- 14555416 TI - Final report on the safety assessment of triacetin. AB - Triacetin, also known as Glyceryl Triacetate, is reported to function as a cosmetic biocide, plasticizer, and solvent in cosmetic formulations, at concentrations ranging from 0.8% to 4.0%. It is a commonly used carrier for flavors and fragrances. Triacetin was affirmed as a generally recognized as safe (GRAS) human food ingredient by the Food and Drug Administration (FDA). Triacetin was not toxic to animals in acute oral or dermal exposures, nor was it toxic in short-term inhalation or parenteral studies, and subchronic feeding and inhalation studies. Triacetin was, at most, slightly irritating to guinea pig skin. However, in one study, it caused erythema, slight edema, alopecia, and desquamation, and did cause some irritation in rabbit eyes. Triacetin was not sensitizing in guinea pigs. Triacetin was not an irritant or a sensitizer in a clinical maximization study, and only very mild reactions were seen in a Duhring chamber test using a 50% dilution. In humans, Triacetin reportedly has caused ocular irritation but no injury. Triacetin was not mutagenic. Although there were no available reproductive and developmental toxicity data, Triacetin was quickly metabolized to glycerol and acetic acid and these chemicals were not developmental toxins. Reports of 1,2-glyceryl diesters, which may be present in Triacetin, affecting cell growth and proliferation raised the possibility of hyperplasia and/or tumor promotion. The Cosmetic Ingredient Review (CIR) Expert Panel concluded, however, that the effects of 1,2-glyceryl diesters on cell growth and proliferation require longer ester chains on the glycerin backbone than are present when acetic acid is esterified with glycerin, as in Triacetin. On the basis of the available information, the CIR Expert Panel concluded that Triacetin is safe as used in cosmetic formulations. PMID- 14555417 TI - Final report on the safety assessment of stearoxy dimethicone, dimethicone, methicone, amino bispropyl dimethicone, aminopropyl dimethicone, amodimethicone, amodimethicone hydroxystearate, behenoxy dimethicone, C24-28 alkyl methicone, C30 45 alkyl methicone, C30-45 alkyl dimethicone, cetearyl methicone, cetyl dimethicone, dimethoxysilyl ethylenediaminopropyl dimethicone, hexyl methicone, hydroxypropyldimethicone, stearamidopropyl dimethicone, stearyl dimethicone, stearyl methicone, and vinyldimethicone. AB - Dimethicone is a fluid mixture of fully methylated linear siloxane polymers end blocked with trimethylsiloxy units. Methicone is a linear monomethyl polysiloxane. The other dimethicones and methicones covered in this review are siloxane polymers of Dimethicone and Methicone. Most of these ingredients function as conditioning agents in cosmetic formulations at current concentrations of use of < or =15%. Clinical and animal absorption studies reported that Dimethicone was not absorbed following oral or dermal exposure. Dimethicone, Methicone, and Vinyldimethicone were not acutely toxic following oral exposure. No adverse reactions were found in rabbits following short-term dermal dosing with 6% to 79% Dimethicone, yet adverse effects were noted with a hand cream formulation containing 1% Dimethicone, suggesting something else in the preparation was toxic. Mice and rats were dosed for 90 days with up to 10% Dimethicone without adverse effect. Dimethicone did not produce adverse effects in acute and short-term inhalation-route studies, Methicone and Vinyldimethicone were negative in acute exposure studies using rats, but Hexyl Methicone was toxic to rats at 5 mg/L delivered in small particle (mean diameter of 0.29 micro) aerosols. Most dermal irritation studies using rabbits classified Dimethicone as a minimal irritant. Dimethicone (tested undiluted and at 79%) was not a sensitizer in four assays using mice and guinea pigs. It was not a sensitizer at 5.0% in a clinical repeated insult patch test using 83 panelists. Most ocular irritation studies using rabbits classified Dimethicone as a mild to minimal irritant. Dimethicone was tested in numerous oral-dose (using rats) and dermal dose (using rats, rabbits, and monkeys) reproductive and developmental toxicity studies. In a few studies, treated males had significantly decreased body weight and/or decreased testes or seminal vesicles weights. No treatment-related adverse findings were noted in dosed pregnant females or fetuses. Dimethicone was negative in all genotoxicity assays. It was negative in both an oral (tested at 91%) and dermal (tested at an unknown concentration) dose carcinogenicity assay using mice. The Cosmetic Ingredient Review (CIR) Expert Panel considered it unlikely that any of these polymers would be significantly absorbed into the skin due to their large molecular weight. Although adverse effects were noted in one inhalation study with small aerosol particles, the expected particle sizes for cosmetic products would primarily be in the range of 60 to 80 micro, and less than 1% would be under 10 micro, which is an upper limit for respirable particles. Overall, the safety test data support the safety of these ingredients at the concentrations they are known to be used in cosmetic formulations. Accordingly, the CIR Expert Panel was of the opinion that Stearoxy Dimethicone, Dimethicone, Methicone, Amino Bispropyl Dimethicone, Aminopropyl Dimethicone, Amodimethicone, Amodimethicone Hydroxystearate, Behenoxy Dimethicone, C24-28 Alkyl Methicone, C30-45 Alkyl Methicone, C30-45 Alkyl Dimethicone, Cetearyl Methicone, Cetyl Dimethicone, Dimethoxysilyl Ethylenediaminopropyl Dimethicone, Hexyl Methicone, Hydroxypropyldimethicone, Stearamidopropyl Dimethicone, Stearyl Dimethicone, Stearyl Methicone, and Vinyldimethicone are safe as used in cosmetic formulations. PMID- 14555418 TI - Final report on the amended safety assessment of sodium polynaphthalenesulfonate and sodium naphthalenesulfonate. AB - Sodium Polynaphthalenesulfonate (SPNS) and Sodium Naphthalenesulfonate (SNS) are sodium salts of naphthalene sulfonic acid. SPNS was used as an emulsion stabilizer, surfactant--hydrotrope, and/or surfactant--suspending agent at concentrations between 0.1% and 0.4%, in a wide range of products, including one lipstick. SNS is described as a surfactant--hydrotrope; no current uses were reported, but information was provided indicating that use concentrations would be typically below 2%. SNS is manufactured by reacting naphthalene with sulfuric acid to produce a sulfonic acid, which is then reacted with sodium hydroxide to produce the final product. The polymer form uses the sulfonic acid intermediate in a reaction with formaldehyde and water under conditions of heat and pressure to form the polymer sulfonic acid form, to which sodium hydroxide is added to make the final SPNS. The residue level of formaldehyde was 0.09%. Only around 1% of SNS in a 1-mg/ml solution applied to porcine skin penetrated the skin after 24 h, a similar amount was found noncovalently bound to the skin, and the concentration of material applied to the surface of the skin was largely unchanged. Both chemicals were not toxic in acute oral or dermal studies. In a subchronic oral toxicity study in rats, the effects noted were increases in urinary sugar in females and urine protein concentrations in males. Although undiluted SPNS was not a significant eye irritant in rabbits, undiluted SNS was a moderate eye irritant in rabbits. At 2%, SNS was a minimal eye irritant in rabbits. Undiluted SNS was at most a mild irritant in Guinea pigs, and was nonirritating at 20% and 2%. In a delayed contact hypersensitivity test in Guinea pigs, 30% SNS used in the induction phase and in the challenge phase produced no reactions. In a Guinea pig maximization test, 1% SNS used with Freund's complete adjuvant (FCA) injected in the initial sensitization, 50% SNS applied topically in the second sensitization, and up to 30% SNS applied topically in the challenge phase did not produce any irritation or sensitization. Both ingredients were negative in Ames mutagenesis assays. In clinical studies, SNS was neither an irritant (tested up to 2%), cumulative irritant (tested up to 1%), nor a sensitizer (tested up to 1%). The Panel considered the low penetration in concert with the low concentrations of use of these ingredients and the absence of significant overall toxicity and the limited negative genotoxicity findings sufficient to support a conclusion that SNS and SPNS are safe as used in cosmetic formulations intended to be applied to the skin. Use of SPNS in a lipstick formulation, was not considered to be different from application to the skin in that the barrier properties of the skin do not apply when these ingredients may contact mucous membranes or may be ingested. Accordingly, the Panel concluded that the available data are insufficient to support the safety of SNS and SPNS in cosmetic formulations that may contact mucous membranes or be ingested. The additional data needed to make a safety assessment for these uses include dermal reproductive and developmental toxicity data and one genotoxicity assay in a mammalian system, and if that study is positive, then a 2-year dermal carcinogenicity study using National Toxicology Program (NTP) methods may be needed. PMID- 14555419 TI - Final report on the amended safety assessment of diisopropyl dimer dilinoleate, dicetearyl dimer dilinoleate, diisostearyl dimer dilinoleate, dioctyl dimer dilinoleate, dioctyldodecyl dimer dilinoleate, and ditridecyl dimer dilinoleate. AB - Diisopropyl Dimer Dilinoleate, Dicetearyl Dimer Dilinoleate, Diisostearyl Dimer Dilinoleate, Dioctyl Dimer Dilinoleate, Dioctyldodecyl Dimer Dilinoleate, and Ditridecyl Dimer Dilinoleate are diesters of their respective alcohols and dilinoleic acid. They function as skin-conditioning agents in a variety of cosmetic products at concentrations around 10%, but may be used at concentrations up to 53% in lipsticks. These ingredients do not absorb radiation in the ultraviolet (UV) UVA or UVB range and the only impurities expected are <0.5% dilinoleic acid, <0.1% isopropyl alcohol or <1% isostearyl alcohol, and/or small amounts of dilinoleic acid and cetearyl alcohol or octyldodecanol, depending on which diester is used. The potential skin penetration of these ingredients was evaluated using an estimate of the octanol/water partition coefficient (logP of 17.7) based on the structure of Diisopropyl Dimer Dilinoleate. This is consistent with the insolubility of these ingredients in water. Safety test data on dilinoleic acid (no adverse effects) were considered relevant because dilinoleic acid is a component of these diesters and a likely breakdown product. The acute oral and dermal LD(50) values for rats of Diisopropyl, Diisostearyl, and Dioctyldodecyl Dimer Dilinoleate were >5.0 g/kg. In a subchronic feeding study, macrophage aggregation was seen in the mesenteric lymph node at the lowest dose level (0.1% in the diet). These ingredients did not produce skin or ocular irritation in animal tests, nor were they comedogenic. Ames testing, clastogenesis in human lymphocytes in culture, and L5178Y mouse lymphoma cell forward mutations were all negative, indicating no dilinoleic acid genotoxicity. No carcinogenicity or reproductive/developmental toxicity data were available; however, structural alerts that would suggest a mutagenic or carcinogenic risk are absent. Significant reproductive/developmental toxicity or other systemic toxicity is not expected with these ingredients because they remain on the skin surface. In clinical studies, cosmetic formulations containing these ingredients did not produce skin irritation or sensitization, although one report of sensitization to dilinoleic acid appeared in the case literature. The Panel did note that the concentration of use of Diisopropyl Dimer Dilinoleate was reportedly as high as 53% in lipsticks, but that the highest concentration tested for irritation/sensitization is 27%. Given the size of these molecules, their relative insolubility in water, their lipophilic nature, and the absence of any significant case reports of allergic reactions, a use concentration of 53% is not likely to be associated with any adverse effects. Accordingly, these diesters were considered safe as used in cosmetic products. PMID- 14555420 TI - Final report on the safety assessment of sodium sulfite, potassium sulfite, ammonium sulfite, sodium bisulfite, ammonium bisulfite, sodium metabisulfite and potassium metabisulfite. AB - Sodium Sulfite, Ammonium Sulfite, Sodium Bisulfite, Potassium Bisulfite, Ammonium Bisulfite, Sodium Metabisulfite, and Potassium Metabisulfite are inorganic salts that function as reducing agents in cosmetic formulations. All except Sodium Metabisulfite also function as hair-waving/straightening agents. In addition, Sodium Sulfite, Potassium Sulfite, Sodium Bisulfite, and Sodium Metabisulfite function as antioxidants. Although Ammonium Sulfite is not in current use, the others are widely used in hair care products. Sulfites that enter mammals via ingestion, inhalation, or injection are metabolized by sulfite oxidase to sulfate. In oral-dose animal toxicity studies, hyperplastic changes in the gastric mucosa were the most common findings at high doses. Ammonium Sulfite aerosol had an acute LC(50) of >400 mg/m(3) in guinea pigs. A single exposure to low concentrations of a Sodium Sulfite fine aerosol produced dose-related changes in the lung capacity parameters of guinea pigs. A 3-day exposure of rats to a Sodium Sulfite fine aerosol produced mild pulmonary edema and irritation of the tracheal epithelium. Severe epithelial changes were observed in dogs exposed for 290 days to 1 mg/m(3) of a Sodium Metabisulfite fine aerosol. These fine aerosols contained fine respirable particle sizes that are not found in cosmetic aerosols or pump sprays. None of the cosmetic product types, however, in which these ingredients are used are aerosolized. Sodium Bisulfite (tested at 38%) and Sodium Metabisulfite (undiluted) were not irritants to rabbits following occlusive exposures. Sodium Metabisulfite (tested at 50%) was irritating to guinea pigs following repeated exposure. In rats, Sodium Sulfite heptahydrate at large doses (up to 3.3 g/kg) produced fetal toxicity but not teratogenicity. Sodium Bisulfite, Sodium Metabisulfite, and Potassium Metabisulfite were not teratogenic for mice, rats, hamsters, or rabbits at doses up to 160 mg/kg. Generally, Sodium Sulfite, Sodium Metabisulfite, and Potassium Metabisulfite were negative in mutagenicity studies. Sodium Bisulfite produced both positive and negative results. Clinical oral and ocular-exposure studies reported no adverse effects. Sodium Sulfite was not irritating or sensitizing in clinical tests. These ingredients, however, may produce positive reactions in dermatologic patients under patch test. In evaluating the positive genotoxicity data found with Sodium Bisulfite, the equilibrium chemistry of sulfurous acid, sulfur dioxide, bisulfite, sulfite, and metabisulfite was considered. This information, however, suggests that some bisulfite may have been present in genotoxicity tests involving the other ingredients and vice versa. On that basis, the genotoxicity data did not give a clear, consistent picture. In cosmetics, however, the bisulfite form is used at very low concentrations (0.03% to 0.7%) in most products except wave sets. In wave sets, the pH ranges from 8 to 9 where the sulfite form would predominate. Skin penetration would be low due to the highly charged nature of these particles and any sulfite that did penetrate would be converted to sulfate by the enzyme sulfate oxidase. As used in cosmetics, therefore, these ingredients would not present a genotoxicity risk. The Cosmetic Ingredient Review Expert Panel concluded that Sodium Sulfite, Potassium Sulfite, Ammonium Sulfite, Sodium Bisulfite, Ammonium Bisulfite, Sodium Metabisulfite, and Potassium Metabisulfite are safe as used in cosmetic formulations. PMID- 14555424 TI - New uses for lithium and anticonvulsants. AB - Lithium and several anticonvulsant medications play an important role in the treatment of bipolar mood disorder. Clinical observations and open clinical trials are beginning to suggest additional therapeutic roles for these drugs. The largest number of reports indicate a monotherapy antidepressant effect for lithium and for lamotrigine. There is also strong evidence for lithium's role in augmenting antidepressant therapy, with reports also suggesting an augmenting role for divalproex and for lamotrigine. The potential usefulness of these medications for 21 other conditions has been reported in the literature, primarily from small case series or open-label, uncontrolled trials. PMID- 14555425 TI - Pretreatment episode count and response to lithium treatment in manic-depressive illness. AB - Factors that reliably predict treatment response in bipolar disorder are much needed, particularly since no available treatment routinely affords complete protection from future illness. The number of pretreatment episodes (PTEs) is a proposed predictor, but its value remains uncertain. We therefore reviewed available research on this topic. Based on a computer-assisted search of the literature, we identified 28 reports providing data on response to lithium treatment and on history of past illness. We evaluated their methods and findings to test the hypothesis that greater PTE count predicts inferior clinical response to lithium. Most studies (68%) found no support for the predicted relationship, and those that did or did not find the hypothesized relationship differed nonsignificantly in ratings of overall study quality and individual factors, including study size, previous lithium use, diagnostic criteria, and outcome measures. The concept that PTE count strongly and consistently predicts inferior clinical response to lithium treatment in manic-depressive disorders is not supported by available research, thus adding to emerging uncertainties about the relationship of past history and later course of these often severe, disabling, and life-threatening illnesses. PMID- 14555426 TI - Clinical use of anticonvulsants: a neurologist's perspective. AB - Seizures may have an acute, reversible cause; antiepileptic drugs (AEDs) are not needed for every patient who presents with a seizure. The neurologist's decision concerning whether to initiate drug therapy depends on verifying that one or more seizures has occurred, on determining the likelihood of recurrence, and on balancing the risks of future seizures against the risks of treatment. Choice of AED is guided by classification of the seizure type and epilepsy syndrome, and by assessment of the likelihood of successful use based on patient characteristics such as age, childbearing potential, and comorbidities, balanced against drug characteristics such as pharmacokinetic profile and common side effects. Dose adjustment depends on communication between the physician and patient regarding seizure control and adverse drug effects. Patients who do not respond adequately to the first two or three appropriate AEDs should be referred for a comprehensive evaluation to verify the diagnosis and determine suitability for alternative treatments such as epilepsy surgery. PMID- 14555427 TI - Clinical significance of brain white matter hyperintensities in young adults with psychiatric illness. AB - Magnetic resonance imaging (MRI) provides detailed images of brain anatomy, with especially clear definition of gray and white matter structures. Several brain MRI studies have suggested that adults with bipolar disorder (BD) are more likely to have "white matter hyperintensities" (WMH) than adults without BD. The disproportionately greater frequency of these lesions in otherwise physically healthy patients suggests that the illness itself, or treatments used to control the illness, may be risk factors for the development of white matter changes. Similarly, WMH may be an etiological factor for some types of BD. In addition to reviewing the relevant literature, this research study attempted to determine whether lithium treatment is associated with an increased prevalence of WMH in young adults with psychiatric illness. To test this hypothesis, we evaluated over 600 brain MRI scans from inpatients at McLean Hospital, Belmont, Massachusetts. We controlled for possible confounding variables such as age, vascular disease, substance abuse, and markers of illness severity. We found that individuals with BD were no more likely to have WMH than other psychiatric patients. Lithium use was nonsignificantly associated with the presence of WMH. A multivariate regression model for the presence of WMH showed that heart disease, female gender, and multiple psychiatric admissions were significant predictors of WMH. This study does not support previous findings that BD, compared to other psychiatric illnesses, was associated with increased risk of WMH. Lithium use may be subtly associated with WMH. Our results are consistent with previous research that found an association between cardiovascular disease, advanced age, and the presence of WMH, though our analysis appears to be unique in its inclusion of cardiovascular disease as a risk factor in young adults with psychiatric illness. PMID- 14555428 TI - A beautiful mind and insulin coma: social constraints on psychiatric diagnosis and treatment. PMID- 14555429 TI - Cristobalite contents of airborne respirable dust in the diatomaceous earth industry in California. PMID- 14555430 TI - Evaluation of elevated blood lead in a commercial painter. PMID- 14555431 TI - Health Effects of Mineral Oil Mist and Metalworking Fluids Symposium. October 2 4, 2002, Cincinnati, Ohio, USA. Proceedings. PMID- 14555432 TI - European hazard classification advice for crude oil-derived lubricant base oils compared with the proposed mineral oil mist TLV. AB - The notice of intended change for the threshold limit value (TLV) for mineral oil mist contains a notation for human carcinogenicity. A description is provided of the current European regulatory approach used to distinguish between carcinogenic and non-carcinogenic mineral base oils on the basis of oil refining process and chemical marker information. This approach has proven effective in creating a market situation in the countries of the European Union where many customers require severely refined, non-carcinogenic oils. It is recommended that ACGIH consolidate the distinction between poorly and severely refined base oils in the recommended TLV for mineral oil mist and use different toxicological considerations to derive exposure control guidelines. PMID- 14555433 TI - Metalworking fluids: oil mist and beyond. AB - This article is based upon my own experiences with metalworking fluids and the adverse health effects and medical conditions associated with exposure to metalworking fluids. I have researched and witnessed the benefits that can be achieved when metalworking fluids are properly maintained and managed. My experiences have provided insight into how a shop operates, including comprehension of the equipment used, processes, mist generating points, engineering controls currently being adopted, and procedures that are used to maintain metalworking fluids. I have been able to share my personal experiences with the country's leading experts in the field of metalworking fluids. I have presented my insights on the topic in Washington, D.C., to the Standard Advisory Committee of OSHA, as well as at many other conferences nationwide. I have provided awareness training for a number of union and nonunion workers. Being a part of developing successful metal removal fluid programs, I realize the importance of transferring and sharing information. Many times an organization is not fully aware of certain conditions and how to combat them. My mission and intent is to properly educate those who are exposed to the harm that metalworking fluids can invoke and to inform those involved of the possible methods of reducing long- and short-term risk. One thing that must be kept in mind is the way we view these fluids. Many shops categorize the fluids as a type of "operating expense" when they should actually be seen as a sort of investment. Just as performing a scheduled maintenance on a machine promises the best possible longevity of that machine, the upkeep of metalworking fluid also provides longer "tool life." Monitoring and maintaining the fluids also provides for more effective and efficient productivity. If we fail to consider that proper management of the fluids can cut cost dramatically, then we will miss out on the financial impact they can have on a company. Try looking at the fluids as a liquid tool. Doing so I believe will bring a better understanding of the value of a successful metalworking fluids program. With this new understanding, it can be seen just who must play a role in the management of metalworking fluids. The employees who deal with the daily tasks involving the coolant play a major part. They are on the floor where these metalworking fluids are being used. In many shops, it is assumed that the environmental health & safety departments are responsible for standard operating procedures and management of fluids. The EH&S department should only be responsible for the protection from exposure and the transfer of information regarding policy and procedure to their employees. Not all shops have the resources required to develop and implement the proper standard operating procedure. Therefore, we must understand that what is feasible for one may not be for another. Companies that lack the sufficient resources should not be neglected. It is crucial that awareness of proper standard operating procedure is shared with everyone involved with the fluids in order to provide proper metalworking fluids management. Fluids are as dynamic as the formulations themselves (complex & dynamic). These fluids can quickly become contaminated with foreign materials and chemicals, thereby become aerosolized into mist. With proper education and training, one will be able to control what gets aerosolized. PMID- 14555434 TI - An evaluation of ASTM method P-42-97 for sampling and analysis of metalworking fluids. PMID- 14555435 TI - Review of respiratory morbidity from occupational exposure to oil mists. AB - The American Conference of Governmental Industrial Hygienists (ACGIH) has recommended that the threshold limit value (TLV) for mineral oil mists be dramatically lowered, based on epidemiological evidence of respiratory health effects among machinists exposed to various metalworking fluid mists. A review of the literature regarding respiratory health effects from either metalworking or non-metalworking fluids suggest that machinists may have experienced slightly higher prevalence of common respiratory symptoms and mild and reversible cross shift changes in some measures of pulmonary function. However, the inconsistency and potential for both random and systematic error in this body of literature argue against drawing definitive conclusions. There is also no substantive evidence that any of these effects led to permanent disease or impairment. The most likely causal agents for respiratory effects in these workers are microbial contaminants in water-based metalworking fluids, not straight mineral oils. This is consistent with the epidemic outbreaks of hypersensitivity pneumonitis, bronchitis, and asthma reported at some work sites using water-based metalworking fluids. This highlights the importance of frequent cleaning and fluid changes for metalworking fluid reservoirs, as part of a systematic approach to managing metalworking fluid aerosol exposures. A dramatic drop in the TLV for mineral oil mists would not resolve this problem. PMID- 14555436 TI - Effects of fluid composition on mist composition. AB - In a reported study, mists of selected synthetic metalworking fluids were generated in laboratory experiments by two processes, nebulization (atomization) and air sparging (bubbling). Short-chain fatty acid species were determined by in situ trimethylsilyl derivatization. Comparison of relative amounts of the short chain acids collected from mists generated by nebulization with those generated by sparging showed that the sparged mists had significantly higher amounts of neodecanoic, nonanoic, and dodecanedioic acids. Comparison of the amounts of acids collected by the resin cartridges to amounts found on the filters showed that significant losses of octanoic and isononanoic acids occurred over 8 hours of collection and that only dodecanedioic acid was not lost from the filter over a 22-hour sampling period. In another reported metalworking mist study, contaminants of metalworking fluids, e.g., tramp oils, were shown in laboratory experiments to increase the misting potential of water-based metalworking fluids. Significantly, tramp oil contamination caused less misting in synthetic fluids than soluble and semi-synthetic fluids. PMID- 14555437 TI - Experimental and analytical efforts to characterize cutting fluid mist formation and behavior in machining. AB - The use of cutting fluids in machining operations is being carefully scrutinized by industry for several reasons, including its overall cost in the manufacturing process and its impact on worker health. Given the concerns associated with the use of cutting fluids, a number of experimental and analytical research efforts are being conducted to gain an understanding of the role of these fluids in various machining processes. The knowledge gained by this research will aid in the development and implementation of strategies to reduce or eliminate the negative effects of cutting fluids, while maintaining their beneficial role. This article presents the results of designed experiments focused on determining the significant variables that influence air quality during turning operations, as well as characterize the aerosol emissions associated with wet and dry turning. Air quality is characterized by measuring the mass concentration and particle size distribution of the dust and mist created during a set of machining experiments. The relative importance of vaporization/condensation and atomization as mist-generating mechanisms is also explored. The experiments revealed that spindle speed has a dominating effect on both mist mass concentration and aerodynamic particle size. Analytical models are presented that predict the average droplet size of the mist generated by atomization and are used to investigate droplet size trends for various cutting fluids and machining parameters. The results predicted by the models are consistent with the expected trends. PMID- 14555438 TI - Summary of the findings from the exposure assessments for metalworking fluid mortality and morbidity studies. AB - Since 1985, a number of North American researchers have evaluated the association between worker exposure to metalworking fluids (MWFs) and cancer mortality or respiratory morbidity. The studies have used different methods to measure the MWF aerosol concentration and to evaluate the exposures to the specific components of the MWF aerosol (bacteria, endotoxin, elements, metals, ethanolamines, polyaromatic hydrocarbons). This diversity of approaches makes comparison of study results difficult and has impeded the development of an occupational exposure limit (OEL) for metalworking fluids. This article summarizes the exposures measured in these North American epidemiologic studies by estimating their thoracic and inhalable MWF particulate levels. In addition, issues that must be resolved before a universal sampling and analysis method for MWF can be recommended are reviewed, including: the use of gravimetric versus extractable analysis; whether the analytical limit of detection can support a lower occupational exposure limit; if the volatile components of mineral oils should also be collected; and whether there are components of specific concern in MWFs that should be regulated separately. Finally, recommendations for future directions in MWF exposure assessment and control are suggested. PMID- 14555440 TI - Controlling health risks from workplace exposure to metalworking fluids in the United Kingdom engineering industry. AB - On October 15, 2002, the United Kingdom (UK) Health and Safety Executive (HSE) launched new guidance for the engineering industry, aimed at reducing health risks from metalworking fluids (MWFs). This guidance was the culmination of many years of work on this subject. In the early 1990s, the UK occupational exposure standards (OES) for oil mist were 5 mg/m(-3) 8-hour time-weighted average (TWA), and 10 mg/m(-3) short-term exposure limit (STEL). This was only applicable to highly refined mineral oil mists and there was no exposure limit for water-mix MWFs (emulsions, semi-synthetics, and synthetics). HSE therefore undertook to review the existing exposure limit for neat mineral oil mists (neat oils are fluids that contain highly refined mineral oils and additives, and are used neat without mixing with water) and consider developing one for water-mix MWFs. This led to the development of new air-sampling methods, a comprehensive survey, and the development of new good practice guidance in the place of statutory exposure limits. This new good practice guidance has been endorsed and launched with the help of relevant industry supplier, employer, and employee associations. The guidance builds on the philosophy of tackling health risks as a holistic approach; for example, not just tackling mist control through the use of ventilation, but also fluid selection, fluid delivery, and fluid management (fluid management means to effectively manage all aspects of the fluid, from storage and stock preparation to sump cleaning and fluid disposal, etc.). Tools, such as laminated task sheets, are provided to make it user friendly. It also demonstrates the business benefits from this approach, that managing your MWFs effectively can reduce the incidence of ill health, reduce fluid and disposal costs, increase tool life, and improve machining performance. PMID- 14555439 TI - Comparison of methods for the measurement of mist and vapor from light mineral oil-based metalworking fluids. AB - The measurement of oil mist derived from metalworking fluids formulated with light mineral oils can be highly inaccurate when using traditional filter sampling. This is due to evaporation of oil from the filter. In this work the practicability of an alternative approach measuring total oil mist and vapor was investigated. Combinations of inhalable particle samplers with backup sorbent vapor traps and standard vapor sampling on pumped and diffusive sorbent tubes were evaluated with gravimetric, infrared spectroscopic, and gas chromatographic analytical methods against the performance requirements of European Standard EN 482. An artificial aerosol was used to compare the methods against a reference method of filter sampler in series with three impingers. Multi-orifice samplers were used with standard 8-mm diameter charcoal tubes at 2 L/min without any signs of channelling or significant breakthrough, as were conical inhalable samplers with XAD-2 tubes at 1 L/min. Most combinations of samplers had a bias of less than 3 percent, but solitary pumped charcoal tubes underestimated total oil by 13 percent. Diffusive sampling was affected by impaction of mist particles and condensation of oil vapor. Gravimetric analysis of filters revealed significant potential sample loss during storage, with 4 percent being lost after one day when stored at room temperature and 2 percent when refrigerated. Samples left overnight in the balance room to equilibrate lost 24 percent. Infrared spectroscopy gave more precise results for vapor than gas chromatography (p = 0.002). Gas chromatography was less susceptible to bias from contaminating solvent vapors than infrared spectroscopy, but was still vulnerable to petroleum distillates. Under the specific test conditions (one oil type and mist particle size), all combinations of methods examined complied with the requirements of European Standard EN 484. Total airborne oil can be measured accurately; however, care must be taken to avoid contamination by hydrocarbon solvent vapors during sampling. PMID- 14555441 TI - Control methods for mineral oil mists. AB - Effective mist collection is important, but it is not the only determinant of mist concentration in plant air. Oil-based metalworking fluids such as straight and soluble oils contain semivolatile hydrocarbons. When these fluids form a mist, their semivolatile components partition between the vapor and mist phases depending on the makeup of the mist and on local conditions. This article addresses the relationship between the concentrations of semivolatile hydrocarbons in the vapor and mist phases using theory for partitioning developed in the field of atmospheric chemistry. Mist can be removed effectively in a collector that uses a HEPA filter as its final collection stage. Acceptable HEPA lifetime requires effective upstream stages that reduce mist loading to the HEPA; furthermore, acceptable HEPA performance requires that it be installed and maintained properly. Collectors designed to remove mist do not remove vapor, and as collector exhaust mixes into cooler plant air that already contains some mist, vapor from the collector can repartition to increase the mist concentration in the plant. Assessing the effect of vapor-to-mist repartitioning is complicated; however, repartitioning may be important for many of the compounds contained in oil-based metalworking fluids. Conditions that minimize vapor-to-mist repartitioning, such as ventilating the plant with clean outdoor air, increasing plant temperature, or controlling the release of vapor, may also be expensive, uncomfortable to plant occupants, or impractical from an engineering standpoint. As a result, very low mist concentrations in plant air may be difficult to attain. PMID- 14555442 TI - Petroleum mineral oil refining and evaluation of cancer hazard. AB - Petroleum base oils (petroleum mineral oils) are manufactured from crude oils by vacuum distillation to produce several distillates and a residual oil that are then further refined. Aromatics including alkylated polycyclic aromatic compounds (PAC) are undesirable constituents of base oils because they are deleterious to product performance and are potentially carcinogenic. In modern base oil refining, aromatics are reduced by solvent extraction, catalytic hydrotreating, or hydrocracking. Chronic exposure to poorly refined base oils has the potential to cause skin cancer. A chronic mouse dermal bioassay has been the standard test for estimating carcinogenic potential of mineral oils. The level of alkylated 3-7 ring PAC in raw streams from the vacuum tower must be greatly reduced to render the base oil noncarcinogenic. The processes that can reduce PAC levels are known, but the operating conditions for the processing units (e.g., temperature, pressure, catalyst type, residence time in the unit, unit engineering design, etc.) needed to achieve adequate PAC reduction are refinery specific. Chronic dermal bioassays provide information about whether conditions applied can make a noncarcinogenic oil, but cannot be used to monitor current production for quality control or for conducting research or developing new processes since this test takes at least 78 weeks to conduct. Three short-term, non-animal assays all involving extraction of oil with dimethylsulfoxide (DMSO) have been validated for predicting potential carcinogenic activity of petroleum base oils: a modified Ames assay of a DMSO extract, a gravimetric assay (IP 346) for wt. percent of oil extracted into DMSO, and a GC-FID assay measuring 3-7-ring PAC content in a DMSO extract of oil, expressed as percent of the oil. Extraction with DMSO concentrates PAC in a manner that mimics the extraction method used in the solvent refining of noncarcinogenic oils. The three assays are described, data demonstrating the validation of the assays are shown, and test results of currently manufactured base oils are summarized to illustrate the general lack of cancer hazard for the base oils now being manufactured. PMID- 14555443 TI - Updated epidemiology of workers exposed to metalworking fluids provides sufficient evidence for carcinogenicity. AB - The 1998 National Institute for Occupational Safety and Health (NIOSH) criteria document for metalworking fluids (MWF) is the most recent authoritative review of mortality studies of workers with these exposures. NIOSH concluded that substantial evidence exists for increased risk of cancer at several sites (larynx, rectum, pancreas, skin, scrotum, and bladder) among workers exposed to MWF before the mid-1970s, and that evidence is equivocal for cancer at several other sites, including stomach, esophagus, lung, prostate, brain, colon, and hematopoietic system. The UAW believes that systematic analysis of that body of data makes a much stronger case for stomach cancer related to MWF exposure. Since the Criteria document, the mortality experience of three of the cohorts reviewed has been either updated or reanalyzed. These updates strengthen the evidence for increased mortality from stomach and liver cancer, and non-malignant respiratory disease associated with exposure to water-based metalworking fluids. Additional toxicological data providing clear evidence for carcinogenicity of diethanolamine, a widely used ingredient, also increases the biological plausibility of these findings. Despite changes in composition of MWFs with time, and reduced exposure levels, these data contradict the notion that cancer risks have been eliminated. PMID- 14555444 TI - Epidemiologic evidence on the carcinogenicity of metalworking fluids. AB - The purpose of this review is to organize and evaluate the epidemiologic evidence regarding the potential carcinogenicity of metalworking fluids. Published literature was initially examined to identify the key contributions, with a strong emphasis on the series of studies by Eisen et al. A key challenge to addressing the issue is the diversity of metalworking fluids, additives, and by products produced in use, along with the notable changes in the composition and use of such agents over time. Although several smaller cohort studies provided useful data on this issue through the 1980s, the Eisen et al. studies offer unique information given the size of the cohort, sophistication in exposure assessment, and detailed analysis of cancer mortality risks within the cohort as a function of estimated exposure. The most notable associations, based on precision, magnitude, and evidence for increasing risk with increasing exposure are those between straight metalworking fluids and both rectal and laryngeal cancer, as well as soluble metalworking fluids and laryngeal cancer. Further progress will require additional studies of the scale of Eisen et al.'s as well as a more systematic approach to integrating information from toxicology and industrial hygiene into the interpretation of the epidemiologic literature. PMID- 14555445 TI - Respiratory toxicology of mineral oils in laboratory animals. AB - Early subchronic and chronic inhalation toxicology studies on various petroleum mineral oils and formulated lubricants supported the ACGIH TLV of 5 mg/m(3) for mineral oil mist. Additional subchronic studies with aerosolized mineral base oils and lubricants during the last 15 years demonstrated that exposures to aerosols of mineral base oils (often >100 mg/m(3)) resulted mainly in concentration-related accumulation in the lung of alveolar macrophages laden with oil droplets. Inflammatory cells were observed with higher aerosol concentrations, consistent with the clinical literature from highly exposed workers. These pulmonary changes appeared to be a nonspecific response to the presence of deposited aerosol. Studies on sensory irritation during exposures of lab animals to mineral oils indicate possible effects only with very high aerosol concentrations. Coupled with changes in refining to remove carcinogenic polynuclear aromatic hydrocarbons from mineral oils several decades ago, this information indicates that current aerosols of mineral oils have a profile of low toxicity from acute to long-term exposures. Available information suggests that additives in some formulated products and/or maintenance of mineral-based metalworking fluids may play a much more significant role in potential health effects. PMID- 14555446 TI - Inpatient hospital admission rates for nonmalignant respiratory disease among workers exposed to metal removal fluids at a U.S. automobile manufacturer. AB - This study was undertaken to determine the impact of exposure to metal removal fluids (MRFs) on the respiratory health of exposed workers. The outcome measure selected was the rate of hospital admissions for nonmalignant respiratory disease episodes as determined from healthcare insurance claims data. A cohort of MRF exposed employees was assembled from 11 manufacturing facilities where MRFs were extensively used in the manufacture of automotive engines, transmissions, and other machined parts. The MRF-exposed cohort included 20,434 employees of such facilities who worked at any time from 1993 through 1997. A non-MRF-exposed cohort was assembled from other employees of the same company during the same time period, but working in warehouse operations and other manufacturing facilities that did not use MRFs or any known respiratory sensitizing agents. The non-exposed cohort included 8681 employees. The crude hospital admission rate for the MRF-exposed cohort was 44 percent higher than that of the non-exposed cohort over the 5-year study period (6.67 vs. 4.62 per 1000 person years at risk, p < 0.05). With age adjustment, the MRF population's rate was still 35 percent higher, and still statistically significant. A nested case-control study was also conducted to determine whether the risk of hospital admission increased with the level of MRF exposure in the population working in MRF plants. The industrial hygiene reconstruction found the levels of exposures of both cases and controls to be very low, with the vast majority of study subjects (more than 90%) having exposures of less than 0.5 mg/m(3). The case-control study did not find any association between increased levels of MRF exposure and risk of hospitalization. The study did document an elevated risk of hospitalization among a sizable population employed in manufacturing operations where MRFs are used. PMID- 14555447 TI - Investigating respiratory responses to metalworking fluid exposure. AB - In response to worker and union representative concerns, the association between metalworking fluid exposure and respiratory symptoms was investigated in a cross sectional survey, in a large automotive machining location. A self-administered respiratory symptom-screening questionnaire was sent to 2935 current employees. MWF exposure levels were assigned to respondents on a departmental basis based on average and "peak" area aerosol measurements. MWF exposure, years in the plant, and smoking status were regressed on presence or absence of daily or weekly respiratory symptoms, as well as upper and lower respiratory symptom groupings derived from principal components factor analysis. The response rate was 81 percent. Symptom prevalence was high: 29 percent of subjects reported weekly or daily phlegm; 23 percent, dry cough; 42 percent, runny or plugged nose. Average aerosol concentration in departments with exposure ranged from 0.02 to 0.84 mg/m(3), and peak levels from 0.02 to 2.85 mg/m(3). Average exposures ranging from 0.25 to 0.84 mg/m(3), as compared to exposures in the range of 0.02 to 0.09 mg/m(3), were statistically significantly associated with wheezing, chest tightness, sore throat, and hoarse throat, as well as with the upper respiratory symptom grouping. When peak exposure was included in the regression, it exerted a stronger effect than average exposure level on dry cough, phlegm, wheezing, fever/chills, and hoarse throat, as well as on upper and lower respiratory symptom groupings. These effects were independent of smoking status. Exposure symptom trends for the average and peak departmental area concentration categories were statistically significant for the upper and lower respiratory symptom groupings and for most individual symptoms. We have observed an association of increasing upper and lower respiratory symptoms with estimated MWF exposure, measured independently, at average departmental aerosol concentrations well below the NIOSH recommended personal exposure level of 0.5 mg/m(3). The results have been used to prioritize exposure reduction efforts in the workplace. PMID- 14555448 TI - Aerosol mapping of a facility with multiple cases of hypersensitivity pneumonitis: demonstration of mist reduction and a possible dose/response relationship. AB - Recent outbreaks of hypersensitivity pneumonitis (HP) have been associated with exposures to metalworking (MWF) fluid aerosols. Mycobacteria present in mismanaged fluids have been suggested as the likely culprit. In January 2001, three machinists at a machining plant were hospitalized with HP. Subsequently, additional HP cases developed as well as cases of bronchitis and occupational asthma. In October 2001, an aerosol concentration map was constructed to advise on priorities for corrective actions. In April 2002 after installation of mist collectors, a second map was generated. Mist reductions varied from about 75 percent to an increase of about 20 percent. Mist concentrations increased in the areas near an uncontrolled transfer line. Subsequent to the first mapping exercise, the exposures of 30 HP cases were classified as low, medium, and high based on job location and the map. There were 6 cases among low exposure jobs (4% of exposed), 14 cases among medium exposure jobs (19% of exposed), and 10 cases among high exposure jobs (34% of exposed), suggesting a relation between exposure to contaminated metalworking fluid mist and the development of HP. A combination of interventions (fluid management, mist control, improved fresh air ventilation, and medical surveillance/restriction) eliminated any new cases 11 months after the first diagnosis of HP. PMID- 14555449 TI - Evaluation of hypersensitivity pneumonitis among workers exposed to metal removal fluids. AB - Hypersensitivity pneumonitis (HP) was identified among employees in an automobile parts manufacturing facility. Mycobacteria immunogenum (MI) was identified as a metal removal fluid (MRF) contaminant at this facility and had been identified as a contaminant in other facilities where HP had occurred. We therefore questioned whether measurement of MI-specific cell-mediated immunity would be associated with HP in this facility. We also questioned whether measures of cell-mediated immunity would be more informative about the presence of HP than evaluation of serum anti-MI antibody levels. Workers were categorized for exposure and disease status by questionnaire and review of medical records. Cell-mediated immunity to MI was assessed by measuring in vitro secretion of cytokines (interleukin 8, tumor necrosis factor alpha, and interferon-gamma) from peripheral blood mononuclear cells or anticoagulated whole blood induced by culture with MI antigen. Serum antibodies against MI were also measured. Six study participants met our survey definition for HP and 48 did not. As has been reported for various agents causing HP, serum antibody levels against MI were increased in both exposed workers and workers with HP. Serum antibodies did not distinguish between the two. When expressed as a percentage of secretion induced by lipopolysaccharide, MI induced a significant increase in interleukin-8 secretion in exposed participants' whole blood cultures. There were trends for increased MI induced secretion of interferon-gamma by peripheral blood mononuclear cells from both exposed workers and workers with HP. However, these trends did not attain statistical significance. Thus, several measures of immunity to MI distinguished between exposed and unexposed workers but not between workers with and without HP. These evaluations of cell-mediated immunity were not more informative than measurement of serum antibodies. As was done at this facility, institution of a comprehensive safety and health plan for MRF is necessary to eliminate (or minimize) health effects related to occupational exposures in the machining environment. PMID- 14555450 TI - Observations on the relationship between triazines and mycobacteria in metal removal fluids. AB - The purpose of the study was to determine whether hexahydrotriazine ("triazine") bactericides increase the likelihood of elevated mycobacteria growth in metal removal fluids (MRFs). In reaction to an outbreak of hypersensitivity pneumonitis (HP) at a transmission manufacturing facility, machining plants of a large automobile manufacturer were directed to increase the testing of MRF, especially testing for mycobacteria. A working hypothesis was developed that triazine bactericides cause an increase in mycobacteria contamination, which in turn may lead to an increased risk of HP for workers exposed to MRF aerosols. Test results for 277 central MRF systems from nine machining facilities were analyzed for various MRF properties including the presence and type of both bactericide in the MRF formula and bacterial contamination of the MRF. The study included data reflecting usual operating conditions of all of the 277 large MRF systems in the company. The study does not evaluate the effect of tank-side biocide addition. The results show that soluble oils, semi-synthetic, and synthetic MRFs are all capable of supporting mycobacterial growth. The results also demonstrate a highly significant association between routine use of triazine bactericides and mycobacterial contamination of MRF. Based on the hypothesis that mycobacteria are a likely causative agent of hypersensitivity pneumonitis from MRF exposure, it is concluded that elimination of triazines from MRF formulae may be protective against HP in the machining environment. PMID- 14555451 TI - DNA-based methodologies for rapid detection, quantification, and species- or strain-level identification of respiratory pathogens (Mycobacteria and Pseudomonads) in metalworking fluids. AB - Mycobacteria and pseudomonads occurring in modern metalworking fluids (MWF) have been implicated in occupational health hazards as causal agents for hypersensitivity pneumonitis (HP) and other respiratory illnesses in machine workers exposed to these fluids and their aerosols. Unlike the conventional cultural and biochemical methods, which are often slow and ambiguous and detect only culturable cells, DNA-based methods offer a time-saving alternative for reliable detection and identification of both culturable and nonculturable bacteria in MWF and for selective quantification of individual genera of pathogens of interest in these fluids. This is the first report on DNA-based direct detection of mycobacteria and pseudomonads in MWF without culturing. Genus specific PCR approach was successfully applied for screening of field MWF samples originating from different industrial users for detection of mycobacteria or pseudomonads including both culturable and nonculturable cells. PCR in combination with amplicon DNA sequencing led to the identification of Mycobacterium chelonae, Pseudomonas nitroreducens, and an undefined Pseudomonas species from these fluids. Genome fingerprinting by pulsed-field gel electrophoresis (PFGE) on Mycobacterium isolates further showed that the isolates represented three strains of M. chelonae although the possibility of one of the strains being clonal with M. immunogenum cannot be excluded. In parallel efforts, a quantitative competitive PCR method developed based on the Pseudomonas-specific PCR was applied to quantify total P. fluorescens cells in contaminated metalworking fluid and MWF aerosol without culturing. The DNA-based protocols developed in this study will allow rapid screening of field MWF samples for the presence of both culturable and nonculturable cells and thus facilitate effective fluid management and timely exposure assessment. PMID- 14555452 TI - Gold, silver, and bronze: metals, medals, and standards in hypersensitivity pneumonitis. PMID- 14555453 TI - Modeling airway remodeling: the winner by a nose? PMID- 14555454 TI - Neutrophil chemokines in severe exacerbations of chronic obstructive pulmonary disease: fatal chemo-attraction? PMID- 14555455 TI - Airway hyperresponsiveness in asthma: geometry is not everything! PMID- 14555456 TI - Phosphodiesterase 4 inhibitors: antiinflammatory therapy for chronic obstructive pulmonary disease at last? PMID- 14555457 TI - Skeletal muscles in chronic airflow obstruction: why bother? PMID- 14555459 TI - Financial disclosure of reviewers. PMID- 14555458 TI - Pathophysiology and management of pulmonary infections in cystic fibrosis. AB - This comprehensive State of the Art review summarizes the current published knowledge base regarding the pathophysiology and microbiology of pulmonary disease in cystic fibrosis (CF). The molecular basis of CF lung disease including the impact of defective cystic fibrosis transmembrane regulator (CFTR) protein function on airway physiology, mucociliary clearance, and establishment of Pseudomonas aeruginosa infection is described. An extensive review of the microbiology of CF lung disease with particular reference to infection with P. aeruginosa is provided. Other pathogens commonly associated with CF lung disease including Staphylococcal aureus, Burkholderia cepacia, Stenotrophomonas maltophilia, Achromobacter xylosoxidans and atypical mycobacteria are also described. Clinical presentation and assessment of CF lung disease including diagnostic microbiology and other measures of pulmonary health are reviewed. Current recommendations for management of CF lung disease are provided. An extensive review of antipseudomonal therapies in the settings of treatment for early P. aeruginosa infection, maintenance for patients with chronic P. aeruginosa infection, and treatment of exacerbation in pulmonary symptoms, as well as antibiotic therapies for other CF respiratory pathogens, are included. In addition, the article discusses infection control policies, therapies to optimize airway clearance and reduce inflammation, and potential future therapies. PMID- 14555460 TI - Ensuring quality in pharmaceutical studies. PMID- 14555461 TI - Methods for the assessment of endobronchial biopsies in clinical research: application to studies of pathogenesis and the effects of treatment. PMID- 14555462 TI - Calcium and ROS-mediated activation of transcription factors and TNF-alpha cytokine gene expression in macrophages exposed to ultrafine particles. AB - Ultrafine (Uf) particles are a component of particulate air pollution suggested to be responsible for the health effects associated with elevations of this pollutant. We have previously suggested that Uf particles, through the induction of oxidative stress, may induce inflammation in the lung, thus exacerbating preexisting illness in susceptible individuals. Alveolar macrophages are considered to play a key role in particlemediated inflammation and lung disease. The effect of Uf particles on rat alveolar macrophages and human blood monocytes was investigated with reference to the roles of calcium and reactive oxygen species (ROS). TNF-alpha protein release, intracellular calcium concentration, TNF-alpha mRNA expression, and transcription factor activation were studied as end points after treatment of rat alveolar macrophages or peripheral blood monocytes. The calcium channel blocker verapamil, the intracellular calcium chelator BAPTA-AM, the calmodulin inhibitor W-7, and the antioxidants Trolox and Nacystelin (NAL) were included in combination with Uf particles. Verapamil reduced intracellular calcium concentration in rat alveolar macrophages on stimulation with Uf particles. This effect was also apparent with transcription factor AP-1 activation. All antagonists and antioxidants reduced Uf-stimulated nuclear localization of the p50 and p65 subunits of NF-kappaB in human monocytes. Verapamil, BAPTA-AM, and NAL reduced Uf-stimulated TNF-alpha protein release, whereas only verapamil reduced Uf-stimulated mRNA expression in rat alveolar macrophages. In human monocytes, verapamil, Trolox, BAPTA-AM, and W-7 reduced Uf stimulated TNF-alpha protein release. These findings suggest that Uf particles may exert proinflammatory effects by modulating intracellular calcium concentrations, activation of transcription factors, and cytokine production through a ROS-mediated mechanism. PMID- 14555463 TI - TNF-alpha induces a decrease in eNOS promoter activity. AB - We determined whether TNF-alpha induces a decrease in activity of the promoter for the endothelial nitric oxide synthase (eNOS) gene in pulmonary microvessel endothelial monolayers (PMEM). eNOS promoter activity was assessed in PMEM transfected with plasmids coding the wild-type (F1: -1600 nt from transcription start site) and truncated (F2: -1189, F4: -779, F5: -494, F6: -166) human eNOS promoters linked to a luciferase reporter. PMEM lysates were analyzed for the luciferase/galactosidase ratio (Luc/Gal) after incubation with TNF-alpha (50 ng/ml) for 0.5 or 4 h. TNF-alpha caused a decrease in the Luc/Gal ratio in the PMEM transfected with wild-type F1 and truncated F2, F4, and F5 plasmids but not with truncated F6 plasmid. Truncated-promoter analysis indicated the response elements (-370)CACCC, (-231)GATA, and (-186)CACCC may regulate the effect of TNF alpha on the eNOS promoter. DNA-binding activity of (32)P-labeled oligonucleotide probes that span the GATA-binding site ((-239)-[(-231)GATA]-(-219)) and the two different CACCC-binding regions ((-379)-[(-370)CACCC]-(-358) and (-196)-[(-186) CACCC]-(-176)) were assessed using EMSA. In response to TNF-alpha treatment for 4 h, nuclear protein binding to (32)P oligonucleotides was characterized as: 1) a significant increase in binding to (-370)CACCC, 2) a significant decrease in binding to (-231)GATA, and 3) no change in (-186)CACCC binding. EMSA supershift analysis indicated that the transcription factor protein GATA-4 bound to the ( 231)GATA site, and Sp3 bound to the (-370)CACCC site. Our data indicate TNF causes a decrease in eNOS promoter activity that may be mediated by GATA-4 and Sp3. PMID- 14555464 TI - Ammonia assimilation by Saccharomyces cerevisiae. PMID- 14555465 TI - trans and cis splicing in trypanosomatids: mechanism, factors, and regulation. PMID- 14555466 TI - Eukaryotic methyl-CpG-binding domain proteins and chromatin modification. PMID- 14555468 TI - Chromosome pairing does not contribute to nuclear architecture in vegetative yeast cells. AB - There are several reports of a closer-than-random colocalization of homologous chromosomes in the vegetative nuclei of diploid budding yeast. Here, we studied by fluorescence in situ hybridization (FISH) the nuclear distribution of chromosomes and found a slight tendency toward closer proximity between homologous (allelic) loci than between any nonhomologous chromosomal regions. We show that most of this preferential association is not due to vegetative (also known as somatic) pairing but is caused by the polar orientation of interphase chromosomes (Rabl orientation). We quantified the occasional loss of detectable fluorescence signals that is inherent to the FISH method. Signal loss leads to the occurrence of a single signal that may be misinterpreted as the close association of two homologous chromosomal sites. The nuclear distribution of homologous loci, when corrected for the influence of nuclear architecture and methodological faults, was not different or was only marginally different from a random relative positioning as predicted by computer simulation. We discuss here several possibilities for the residual homologous proximity that do not invoke homology-dependent vegetative pairing, and we conclude that, in diploid budding yeast, constitutive vegetative pairing is a negligible factor for the organization of the interphase nucleus. PMID- 14555467 TI - Alpha-pheromone-induced "shmooing" and gene regulation require white-opaque switching during Candida albicans mating. AB - A 14-mer alpha-pheromone peptide of Candida albicans was chemically synthesized and used to analyze the role of white-opaque switching in the mating process. The alpha-pheromone peptide blocked cell multiplication and induced "shmooing" in a/a cells expressing the opaque-phase phenotype but not in a/a cells expressing the white-phase phenotype. The alpha-pheromone peptide induced these effects at 25 degrees C but not at 37 degrees C. An analysis of mating-associated gene expression revealed several categories of gene regulation, including (i) MTL homozygous-specific, pheromone stimulated, switching-independent (CAG1 and STE4); (ii) mating type-specific, pheromone-induced, switching-independent (STE2); and (iii) pheromone-induced, switching-dependent (FIG1, KAR4, and HWP1). An analysis of switching-regulated genes revealed an additional category of opaque-phase specific genes that are downregulated by alpha-pheromone only in a/a cells (OP4, SAP1, and SAP3). These results demonstrate that alpha-pheromone causes shmooing, the initial step in the mating process, only in a/a cells expressing the opaque phenotype and only at temperatures below that in the human host. These results further demonstrate that although some mating-associated genes are stimulated by the alpha-pheromone peptide in both white- and opaque-phase cells, others are stimulated only in opaque-phase cells, revealing a category of gene regulation unique to C. albicans in which alpha-pheromone induction requires the white opaque transition. These results demonstrate that in C. albicans, the mating process and associated gene regulation must be examined within the context of white-opaque switching. PMID- 14555469 TI - D-xylose metabolism in Hypocrea jecorina: loss of the xylitol dehydrogenase step can be partially compensated for by lad1-encoded L-arabinitol-4-dehydrogenase. AB - With the goal of the genetic characterization of the D-xylose pathway in Hypocrea jecorina (anamorph: Trichoderma reesei), we cloned the xdh1 gene, encoding NAD xylitol dehydrogenase, which catalyzes the second step of fungal D-xylose catabolism. This gene encodes a 363-amino-acid protein which has a mass of 38 kDa, belongs to the zinc-containing alcohol dehydrogenase family, exhibits high sequence identity to the published sequences of xylitol dehydrogenases from yeast origins, but contains a second, additional binding site for Zn2+. The enzyme catalyzed the NAD-dependent oxidation of xylitol and D-sorbitol and the NADH dependent reduction of D-xylulose and D-fructose. No activity was observed with NADP, L-arabinose, or L-arabinitol. A single 1.4-kb transcript was formed during growth on xylan, D-xylose, L-arabinose, L-arabinitol and, at a lower abundance, xylitol, D-galactose, galactitol, and lactose but not on D-glucose and glycerol. xdh1 deletion mutants exhibited 50% reduced growth rates on D-xylose, whereas growth rates on xylitol remained unaltered. These mutants contained 30% of the xylitol dehydrogenase activity of the parent strain, indicating the presence of a second xylitol dehydrogenase. This activity was shown to be due to lad1-encoded L arabinitol-4-dehydrogenase, because H. jecorina xdh1 lad1 double-deletion strains failed to grow on D-xylose or xylitol. In contrast, lad1 deletion strains of H. jecorina grew normally on these carbon sources. These results show that H. jecorina contains a single xylitol dehydrogenase which is encoded by xdh1 and is involved in the metabolism of D-xylose and that lad1-encoded L-arabinitol-4 dehydrogenase can compensate for it partially in mutants with a loss of xdh1 function. PMID- 14555470 TI - Nucleosome position-dependent and -independent activation of HIS7 epression in Saccharomyces cerevisiae by different transcriptional activators. AB - ARO4 and HIS7 are two tandemly orientated genes of Saccharomyces cerevisiae that are transcribed into the same direction. The ARO4 terminator and the HIS7 promoter regions are sensitive to Micrococcus nuclease (Mnase) and separated by a positioned nucleosome. The HIS7 promoter is target for the transcription factors Gcn4p and Bas1p/Bas2p that activate its transcription upon amino acid starvation and purine limitation, respectively. Activation of the HIS7 gene by Gcn4p overexpression but not by Bas1p/Bas2p releases an ordered nucleosome distribution to yield increased Mnase sensitivity throughout the intergenic region. This remodeling is SNF2 dependent but mostly GCN5 independent. Accordingly, SNF2 is necessary for the Gcn4p-mediated transcriptional activation of the HIS7 gene. GCN5 is required for activation upon adenine limitation by Bas1p/Bas2p. Our data suggest that activation of HIS7 transcription by Gcn4p and Bas1p/Bas2p is supported by a nucleosome position-dependent and -independent mechanism, respectively. Whereas Gcn4p activation causes Swi/Snf-mediated remodeling of the nucleosomal architecture at the HIS7 promoter, the Bas1p/Bas2p complex presumably activates in combination with Gcn5p-dependent histone acetylation. PMID- 14555472 TI - Conservation of RNase III processing pathways and specificity in hemiascomycetes. AB - Rnt1p, the only known Saccharomyces cerevisiae RNase III endonuclease, plays important functions in the processing of precursors of rRNAs (pre-rRNAs) and of a large number of small nuclear RNAs (snRNAs) and small nucleolar RNAs (snoRNAs). While most eukaryotic RNases III, including the Schizosaccharomyces pombe enzyme Pac1p, cleave double-stranded RNA without sequence specificity, Rnt1p cleavage relies on the presence of terminal tetraloop structures that carry the consensus sequence AGNN. To search for the conservation of these processing signals, I have systematically analyzed predicted secondary structures of the 3' external transcribed spacer (ETS) sequences of the pre-rRNAs and of flanking sequences of snRNAs and snoRNAs from sequences available in 13 other Hemiascomycetes species. In most of these species, except in Yarrowia lipolytica, double-stranded RNA regions capped by terminal AGNN tetraloops can be found in the 3' ETS sequences of rRNA, in the 5'- or 3'-end flanking sequences of sn(o)RNAs, or in the intergenic spacers of polycistronic snoRNA transcription units. This analysis shows that RNase III processing signals and RNase III cleavage specificity are conserved in most Hemiascomycetes species but probably not in the evolutionarily more distant species Y. lipolytica. PMID- 14555471 TI - Chitin synthesis in Saccharomyces cerevisiae in response to supplementation of growth medium with glucosamine and cell wall stress. AB - In Saccharomyces cerevisiae most chitin is synthesized by Chs3p, which deposits chitin in the lateral cell wall and in the bud-neck region during cell division. We have recently found that addition of glucosamine (GlcN) to the growth medium leads to a three- to fourfold increase in cell wall chitin levels. We compared this result to the increases in cellular chitin levels associated with cell wall stress and with treatment of yeast with mating pheromone. Since all three phenomena lead to increases in precursors of chitin, we hypothesized that chitin synthesis is at least in part directly regulated by the size of this pool. This hypothesis was strengthened by our finding that addition of GlcN to the growth medium causes a rapid increase in chitin synthesis without any pronounced change in the expression of more than 6,000 genes monitored with Affymetrix gene expression chips. In other studies we found that the specific activity of Chs3p is higher in the total membrane fractions from cells grown in GlcN and from mutants with weakened cell walls. Sucrose gradient analysis shows that Chs3p is present in an inactive form in what may be Golgi compartments but as an active enzyme in other intracellular membrane-bound vesicles, as well as in the plasma membrane. We conclude that Chs3p-dependent chitin synthesis in S. cerevisiae is regulated both by the levels of intermediates of the UDP-GlcNAc biosynthetic pathway and by an increase in the activity of the enzyme in the plasma membrane. PMID- 14555473 TI - Fission yeast Sap1 protein is essential for chromosome stability. AB - Sap1 is a dimeric sequence-specific DNA binding-protein, initially identified for its role in mating-type switching of the fission yeast Schizosaccharomyces pombe. The protein is relatively abundant, around 10,000 dimers/cell, and is localized in the nucleus. sap1+ is essential for viability, and transient overexpression is accompanied by rapid cell death, without an apparent checkpoint response and independently of mating-type switching. Time lapse video microscopy of living cells revealed that the loss of viability is accompanied by abnormal mitosis and chromosome fragmentation. Overexpression of the C terminus of Sap1 induces minichromosome loss associated with the "cut" phenotype (uncoupling mitosis and cytokinesis). These phenotypes are favored when the C terminus of Sap1 is overexpressed during DNA replication. Fluorescence in situ hybridization experiments demonstrated that the cut phenotype is related to precocious centromere separation, a typical marker for loss of cohesion. We propose that Sap1 is an architectural chromatin-associated protein, required for chromosome organization. PMID- 14555474 TI - Constitutive and hyperresponsive signaling by mutant forms of Saccharomyces cerevisiae amino acid sensor Ssy1. AB - Sensing of extracellular amino acids results in transcriptional induction of amino acid permease genes in yeast. Ssy1, a membrane protein resembling amino acid permeases, is required for signaling but is apparently unable to transport amino acids and is thus believed to be a sensor. By using a novel genetic screen in which potassium uptake was made dependent on amino acid signaling, we obtained gain-of-function mutations in SSY1. Some alleles confer inducer-independent signaling; others increase the apparent affinity for inducers. The results reveal that amino acid transport is not required for signaling and support the notion that sensing by Ssy1 occurs via its direct interaction with extracellular amino acids. PMID- 14555475 TI - Attachment of the ubiquitin-related protein Urm1p to the antioxidant protein Ahp1p. AB - Urm1p is a ubiquitin-related protein that serves as a posttranslational modification of other proteins. Urm1p conjugation has been implicated in the budding process and in nutrient sensing. Here, we have identified the first in vivo target for the urmylation pathway as the antioxidant protein Ahp1p. The attachment of Urm1p to Ahp1p requires the E1 for the urmylation pathway, Uba4p. Loss of the urmylation pathway components results in sensitivity to a thiol specific oxidant, as does loss of Ahp1p, implying that urmylation has a role in an oxidative-stress response. Moreover, treatment of cells with thiol-specific oxidants affects the abundance of Ahp1p-Urm1p conjugates. These results suggest that the conjugation of Urm1p to Ahp1p could regulate the function of Ahp1p in antioxidant stress response in Saccharomyces cerevisiae. PMID- 14555476 TI - Regulation of ENA1 Na(+)-ATPase gene expression by the Ppz1 protein phosphatase is mediated by the calcineurin pathway. AB - Saccharomyces cerevisiae strains lacking the Ppz1 protein phosphatase are salt tolerant and display increased expression of the ENA1 Na(+)-ATPase gene, a major determinant for sodium extrusion, while cells devoid of the similar Ppz2 protein do not show these phenotypes. However, a ppz1 ppz2 mutant displays higher levels of ENA1 expression than the ppz1 strain. We show here that the increased activity of the ENA1 promoter in a ppz1 ppz2 mutant maps to two regions: one region located at -751 to -667, containing a calcineurin-dependent response element (CDRE), and one downstream region (-573 to -490) whose activity responds to intracellular alkalinization. In contrast, the increased ENA1 expression in a ppz1 mutant is mediated solely by an intact calcineurin/Crz1 signaling pathway, on the basis that (i) this effect maps to a single region that contains the CDRE and (ii) it is blocked by the calcineurin inhibitor FK506, as well as by deletion of the CNB1 or CRZ1 gene. The calcineurin dependence of the increased ENA1 expression of a ppz1 mutant would suggest that Ppz1 could negatively regulate calcineurin activity. In agreement with this notion, a ppz1 strain is calcium sensitive, and this mutation does not result in a decrease in the calcium hypertolerance of a cnb1 mutant. It has been shown that ENA1 can be induced by alkalinization of the medium and that a ppz1 ppz2 strain has a higher intracellular pH. However, we present several lines of evidence that show that the gene expression profile of a ppz1 mutant does not involve an alkalinization effect. In conclusion, we have identified a novel role for calcineurin, but not alkalinization, in the control of ENA1 expression in ppz1 mutants. PMID- 14555477 TI - Phosphorylation of the MAPKKK regulator Ste50p in Saccharomyces cerevisiae: a casein kinase I phosphorylation site is required for proper mating function. AB - The Ste50 protein of Saccharomyces cerevisiae is a regulator of the Ste11p protein kinase. Ste11p is a member of the MAP3K (or MEKK) family, which is conserved from yeast to mammals. Ste50p is involved in all the signaling pathways that require Ste11p function, yet little is known about the regulation of Ste50p itself. Here, we show that Ste50p is phosphorylated on multiple serine/threonine residues in vivo. Threonine 42 (T42) is phosphorylated both in vivo and in vitro, and the protein kinase responsible has been identified as casein kinase I. Replacement of T42 with alanine (T42A) compromises Ste50p function. This mutation abolishes the ability of overexpressed Ste50p to suppress either the mating defect of a ste20 ste50 deletion mutant or the mating defect of a strain with a Ste11p deleted from its sterile-alpha motif domain. Replacement of T42 with a phosphorylation-mimetic aspartic acid residue (T42D) permits wild-type function in all assays of Ste50p function. These results suggest that phosphorylation of T42 of Ste50p is required for proper signaling in the mating response. However, this phosphorylation does not seem to have a detectable role in modulating the high-osmolarity glycerol synthesis pathway. PMID- 14555478 TI - Ask10p mediates the oxidative stress-induced destruction of the Saccharomyces cerevisiae C-type cyclin Ume3p/Srb11p. AB - Srb11p-Srb10p is the budding yeast C-type cyclin-cyclin-dependent kinase that is required for the repression of several stress response genes. To relieve this repression, Srb11p is destroyed in cells exposed to stressors, including heat shock and oxidative stress. In the present study, we identified Ask10p (for activator of Skn7) by two-hybrid analysis as an interactor with Srb11p. Coimmunoprecipitation studies confirmed this association, and we found that, similar to Srb11p-Srb10p, Ask10p is a component of the RNA polymerase II holoenzyme. Ask10p is required for Srb11p destruction in response to oxidative stress but not heat shock. Moreover, this destruction is important since the hypersensitivity of an ask10 mutant strain to oxidative stress is rescued by deleting SRB11. We further show that Ask10p is phosphorylated in response to oxidative stress but not heat shock. This modification requires the redundant mitogen-activated protein (MAP) kinase kinase Mkk1/2 but not their normal MAP kinase target Slt2p. Moreover, the other vegetative MAP kinases--Hog1p, Fus3p, or Kss1p--are not required for Ask10p phosphorylation, suggesting the existence of an alternative pathway for transducing the Pkc1p-->Bck1-->Mkk1/2 oxidative stress signal. In conclusion, Ask10p is a new component of the RNA polymerase II holoenzyme and an important regulator of the oxidative stress response. In addition, these results define a new role for the Pkc1p MAP kinase cascade (except the MAP kinase itself) in transducing the oxidative damage signal directly to the RNA polymerase II holoenzyme, thereby bypassing the stress activated transcription factors. PMID- 14555479 TI - A Phytophthora infestans G-protein beta subunit is involved in sporangium formation. AB - The heterotrimeric G-protein pathway regulates cellular responses to a wide range of extracellular signals in virtually all eukaryotes. It also controls various developmental processes in the oomycete plant pathogen Phytophthora infestans, as was concluded from previous studies on the role of the G-protein alpha-subunit PiGPA1 in this organism. The expression of the P. infestans G-protein beta subunit gene Pigpb1 was induced in nutrient-starved mycelium before the onset of sporangium formation. The gene was hardly expressed in mycelium incubated in rich growth medium. The introduction of additional copies of Pigpb1 into the genome led to silencing of the gene and resulted in transformants deficient in PiGPB1. These Pigpb1-silenced mutants formed very few asexual spores (sporangia) when cultured in rye sucrose medium and produced a denser mat of aerial mycelium than the wild type. Partially Pigpb1-silenced mutants showed intermediate phenotypes with regard to sporulation, and a relatively large number of their sporangia were malformed. The results show that PiGPB1 is important for vegetative growth and sporulation and, therefore, for the pathogenicity of this organism. PMID- 14555480 TI - Proteomics of Chlamydomonas reinhardtii light-harvesting proteins. AB - With the recent development of techniques for analyzing transmembrane thylakoid proteins by two-dimensional gel electrophoresis, systematic approaches for proteomic analyses of membrane proteins became feasible. In this study, we established detailed two-dimensional protein maps of Chlamydomonas reinhardtii light-harvesting proteins (Lhca and Lhcb) by extensive tandem mass spectrometric analysis. We predicted eight distinct Lhcb proteins. Although the major Lhcb proteins were highly similar, we identified peptides which were unique for specific lhcbm gene products. Interestingly, lhcbm6 gene products were resolved as multiple spots with different masses and isoelectric points. Gene tagging experiments confirmed the presence of differentially N-terminally processed Lhcbm6 proteins. The mass spectrometric data also revealed differentially N terminally processed forms of Lhcbm3 and phosphorylation of a threonine residue in the N terminus. The N-terminal processing of Lhcbm3 leads to the removal of the phosphorylation site, indicating a potential novel regulatory mechanism. At least nine different lhca-related gene products were predicted by comparison of the mass spectrometric data against Chlamydomonas expressed sequence tag and genomic databases, demonstrating the extensive variability of the C. reinhardtii Lhca antenna system. Out of these nine, three were identified for the first time at the protein level. This proteomic study demonstrates the complexity of the light-harvesting proteins at the protein level in C. reinhardtii and will be an important basis of future functional studies addressing this diversity. PMID- 14555481 TI - Copper response element and Crr1-dependent Ni(2+)-responsive promoter for induced, reversible gene expression in Chlamydomonas reinhardtii. AB - The Cpx1 and Cyc6 genes of Chlamydomonas reinhardtii are activated in copper deficient cells via a signal transduction pathway that requires copper response elements (CuREs) and a copper response regulator defined by the CRR1 locus. The two genes can also be activated by provision of nickel or cobalt ions in the medium. The response to nickel ions requires at least one CuRE and also CRR1 function, suggesting that nickel interferes with a component in the nutritional copper signal transduction pathway. Nickel does not act by preventing copper uptake/utilization because (i) holoplastocyanin formation is unaffected in Ni(2+) treated cells and (ii) provision of excess copper cannot reverse the Ni-dependent activation of the target genes. The CuRE is sufficient for conferring Ni responsive expression to a reporter gene, which suggests that the system has practical application as a vehicle for inducible gene expression. The inducer can be removed either by replacing the medium or by chelating the inducer with excess EDTA, either of which treatments reverses the activation of the target genes. PMID- 14555482 TI - Mechanisms of arsenical and diamidine uptake and resistance in Trypanosoma brucei. AB - Sleeping sickness, caused by Trypanosoma brucei spp., has become resurgent in sub Saharan Africa. Moreover, there is an alarming increase in treatment failures with melarsoprol, the principal agent used against late-stage sleeping sickness. In T. brucei, the uptake of melarsoprol as well as diamidines is thought to be mediated by the P2 aminopurine transporter, and loss of P2 function has been implicated in resistance to these agents. The trypanosomal gene TbAT1 has been found to encode a P2-type transporter when expressed in yeast. Here we investigate the role of TbAT1 in drug uptake and drug resistance in T. brucei by genetic knockout of TbAT1. Tbat1-null trypanosomes were deficient in P2-type adenosine transport and lacked adenosine-sensitive transport of pentamidine and melaminophenyl arsenicals. However, the null mutants were only slightly resistant to melaminophenyl arsenicals and pentamidine, while resistance to other diamidines such as diminazene was more pronounced. Nevertheless, the reduction in drug sensitivity might be of clinical significance, since mice infected with tbat1-null trypanosomes could not be cured with 2 mg of melarsoprol/kg of body weight for four consecutive days, whereas mice infected with the parental line were all cured by using this protocol. Two additional pentamidine transporters, HAPT1 and LAPT1, were still present in the null mutant, and evidence is presented that HAPT1 may be responsible for the residual uptake of melaminophenyl arsenicals. High-level arsenical resistance therefore appears to involve the loss of more than one transporter. PMID- 14555483 TI - A negative regulatory element controls mRNA abundance of the Leishmania mexicana Paraflagellar rod gene PFR2. AB - The Leishmania mexicana PFR2 locus encodes a component of the paraflagellar rod (PFR), a flagellar structure found only in the insect stage of the life cycle. PFR2 mRNA levels are 10-fold lower in the mammalian stage than in the insect stage. Nuclear run-on experiments indicate that the change in PFR2 mRNA abundance is achieved posttranscriptionally. Deletion and block substitution analysis of the entire 1,400-nucleotide 3' untranslated region (UTR) of PFR2C led to the identification of a regulatory element contained within 10 nucleotides of the 3' UTR, termed the PFR regulatory element (PRE), that is necessary for the 10-fold regulation of PFR2 mRNA levels. Comparison of the half-lives of PFR2 transcripts, identical except for the presence or absence of the PRE, revealed that the PRE acts by destabilizing the PFR2 mRNA in amastigotes. The PRE was inserted into a construct which directs the constitutive expression of a chimeric PFR2 transcript. Insertion of the PRE resulted in regulated expression of this transcript, demonstrating that the regulatory element is sufficient for promastigote-specific expression. Since the PRE is present in the 3' UTR of all L. mexicana PFR genes examined so far, we propose that it serves a means of coordinating expression of PFR genes. PMID- 14555484 TI - Candida albicans response regulator gene SSK1 regulates a subset of genes whose functions are associated with cell wall biosynthesis and adaptation to oxidative stress. AB - Ssk1p of Candida albicans is a putative response regulator protein of the Hog1 two-component signal transduction system. In Saccharomyces cerevisiae, the phosphorylation state of Ssk1p determines whether genes that promote the adaptation of cells to osmotic stress are activated. We have previously shown that C. albicans SSK1 does not complement the ssk1 mutant of S. cerevisiae and that the ssk1 mutant of C. albicans is not sensitive to sorbitol. In this study, we show that the C. albicans ssk1 mutant is sensitive to several oxidants, including hydrogen peroxide, t-butyl hydroperoxide, menadione, and potassium superoxide when each is incorporated in yeast extract-peptone-dextrose (YPD) agar medium. We used DNA microarrays to identify genes whose regulation is affected by the ssk1 mutation. RNA from mutant cells (strain CSSK21) grown in YPD medium for 3 h at 30 degrees C was reverse transcribed and then compared with similarly prepared RNA from wild-type cells (CAF2). We observed seven genes from mutant cells that were consistently up regulated (three-fold or greater compared to CAF2). In S. cerevisiae, three (AHP1, HSP12, and PYC2) of the seven genes that were up regulated provide cells with an adaptation function in response to oxidative stress; another gene (GPH1) is regulated under stress conditions by Hog1p. Three other genes that are up regulated encode a cell surface protein (FLO1), a mannosyl transferase (MNN4-4), and a putative two-component histidine kinase (CHK1) that regulates cell wall biosynthesis in C. albicans. Of the down regulated genes, ALS1 is a known cell adhesin in C. albicans. Verification of the microarray data was obtained by reverse transcription-PCR for HSP12, AHP1, CHK1, PYC2, GPH1, ALS1, MNN4-4, and FLO1. To further determine the function of Ssk1p in the Hog1p signal transduction pathway in C. albicans, we used Western blot analysis to measure phosphorylation of Hog1p in the ssk1 mutant of C. albicans when grown under either osmotic or oxidative stress. We observed that Hog1p was phosphorylated in the ssk1 mutant of C. albicans when grown in a hyperosmotic medium but was not phosphorylated in the ssk1 mutant when the latter was grown in the presence of hydrogen peroxide. These data indicate that C. albicans utilizes the Ssk1p response regulator protein to adapt cells to oxidative stress, while its role in the adaptation to osmotic stress is less certain. Further, SSK1 appears to have a regulatory function in some aspects of cell wall biosynthesis. Thus, the functions of C. albicans SSK1 differ from those of S. cerevisiae SSK1. PMID- 14555485 TI - Phospholipid-binding protein Cts1 controls septation and functions coordinately with calcineurin in Cryptococcus neoformans. AB - Cryptococcus neoformans is an opportunistic fungal pathogen that causes life threatening meningoencephalitis in immunocompromised patients. The Ca(2+) calmodulin-activated protein phosphatase calcineurin is necessary for virulence of C. neoformans. Mutants lacking the calcineurin catalytic (Cna1) or regulatory (Cnb1) subunit fail to grow at elevated temperature and are defective in virulence and hyphal elongation. Here we isolated a multicopy suppressor gene, CTS1, which restores growth of a calcineurin mutant strain at 37 degrees C. The CTS1 gene (for calcineurin temperature suppressor 1) encodes a protein containing a C2 domain and a leucine zipper motif that may function as an effector of calcineurin. The CTS1 gene was disrupted by homologous recombination, and cts1 mutants were viable but exhibited defects in cell separation, growth, mating, and haploid fruiting. In addition, cts1 mutants were inviable when calcineurin was mutated or inhibited. Taken together, these findings suggest that calcineurin and Cts1 function in parallel pathways that regulate growth, cell separation, and hyphal elongation. PMID- 14555486 TI - Recapitulation of the sexual cycle of the primary fungal pathogen Cryptococcus neoformans var. gattii: implications for an outbreak on Vancouver Island, Canada. AB - Cryptococcus neoformans is a human fungal pathogen that exists as three distinct varieties or sibling species: the predominantly opportunistic pathogens C. neoformans var. neoformans (serotype D) and C. neoformans var. grubii (serotype A) and the primary pathogen C. neoformans var. gattii (serotypes B and C). While serotypes A and D are cosmopolitan, serotypes B and C are typically restricted to tropical regions. However, serotype B isolates of C. neoformans var. gattii have recently caused an outbreak on Vancouver Island in Canada, highlighting the threat of this fungus and its capacity to infect immunocompetent individuals. Here we report a large-scale analysis of the mating abilities of serotype B and C isolates from diverse sources and identify unusual strains that mate robustly and are suitable for further genetic analysis. Unlike most isolates, which are of both the a and alpha mating types but are predominantly sterile, the majority of the Vancouver outbreak strains are exclusively of the alpha mating type and the majority are fertile. In an effort to enhance mating of these isolates, we identified and disrupted the CRG1 gene encoding the GTPase-activating protein involved in attenuating pheromone response. crg1 mutations dramatically increased mating efficiency and enabled mating with otherwise sterile isolates. Our studies provide a genetic and molecular foundation for further studies of this primary pathogen and reveal that the Vancouver Island outbreak may be attributable to a recent recombination event. PMID- 14555487 TI - Role of second-largest RNA polymerase I subunit Zn-binding domain in enzyme assembly. AB - The second-largest subunits of eukaryal RNA polymerases are similar to the beta subunits of prokaryal RNA polymerases throughout much of their lengths. The second-largest subunits from eukaryal RNA polymerases contain a four-cysteine Zn binding domain at their C termini. The domain is also present in archaeal homologs but is absent from prokaryal homologs. Here, we investigated the role of the C-terminal Zn-binding domain of Rpa135, the second-largest subunit of yeast RNA polymerase I. Analysis of nonfunctional Rpa135 mutants indicated that the Zn binding domain is required for recruitment of the largest subunit, Rpa190, into the RNA polymerase I complex. Curiously, the essential function of the Rpa135 Zn binding domain is not related to Zn(2+) binding per se, since replacement of only one of the four cysteine residues with alanine led to the loss of Rpa135 function. Even more strikingly, replacement of all four cysteines with alanines resulted in functional Rpa135. PMID- 14555488 TI - Engineered control of cell morphology in vivo reveals distinct roles for yeast and filamentous forms of Candida albicans during infection. AB - It is widely assumed that the ability of Candida albicans to switch between different morphologies is required for pathogenesis. However, most virulence studies have used mutants that are permanently locked into either the yeast or filamentous forms which are avirulent but unsuitable for discerning the role of morphogenetic conversions at the various stages of the infectious process. We have constructed a strain in which this developmental transition can be externally modulated both in vitro and in vivo. This was achieved by placing one copy of the NRG1 gene (a negative regulator of filamentation) under the control of a tetracycline-regulatable promoter. This modified strain was then tested in an animal model of hematogenously disseminated candidiasis. Mice injected with this strain under conditions permitting hyphal development succumbed to the infection, whereas all of the animals injected under conditions that inhibited this transition survived. Importantly, fungal burdens were almost identical in both sets of animals, indicating that, whereas filament formation appears to be required for the mortality resulting from a deep-seated infection, yeast cells play an important role early in the infectious process by extravasating and disseminating to the target organs. Moreover, these infecting Candida yeast cells still retained their pathogenic potential, as demonstrated by allowing this developmental transition to occur at various time points postinfection. We demonstrate here the importance of morphogenetic conversions in C. albicans pathogenesis. This engineered strain should provide a useful tool in unraveling the individual contributions of the yeast and filamentous forms at various stages of the infectious process. PMID- 14555489 TI - Sfp1 plays a key role in yeast ribosome biogenesis. AB - Sfp1, an unusual zinc finger protein, was previously identified as a gene that, when overexpressed, imparted a nuclear localization defect. sfp1 cells have a reduced size and a slow growth phenotype. In this study we show that SFP1 plays a role in ribosome biogenesis. An sfp1 strain is hypersensitive to drugs that inhibit translational machinery. sfp1 strains also have defects in global translation as well as defects in rRNA processing and 60S ribosomal subunit export. Microarray analysis has previously shown that ectopically expressed SFP1 induces the transcription of a large subset of genes involved in ribosome biogenesis. Many of these induced genes contain conserved promoter elements (RRPE and PAC). Our results show that activation of transcription from a reporter construct containing two RRPE sites flanking a single PAC element is SFP1 dependent. However, we have been unable to detect direct binding of the protein to these elements. This suggests that regulation of genes containing RRPEs is dependent upon Sfp1 but that Sfp1 may not directly bind to these conserved promoter elements; rather, activation may occur through an indirect mechanism. PMID- 14555490 TI - Bacterial catalase in the microsporidian Nosema locustae: implications for microsporidian metabolism and genome evolution. AB - Microsporidia constitute a group of extremely specialized intracellular parasites that infect virtually all animals. They are highly derived, reduced fungi that lack several features typical of other eukaryotes, including canonical mitochondria, flagella, and peroxisomes. Consistent with the absence of peroxisomes in microsporidia, the recently completed genome of the microsporidian Encephalitozoon cuniculi lacks a gene for catalase, the major enzymatic marker for the organelle. We show, however, that the genome of the microsporidian Nosema locustae, in contrast to that of E. cuniculi, encodes a group II large-subunit catalase. Surprisingly, phylogenetic analyses indicate that the N. locustae catalase is not specifically related to fungal homologs, as one would expect, but is instead closely related to proteobacterial sequences. This finding indicates that the N. locustae catalase is derived by lateral gene transfer from a bacterium. The catalase gene is adjacent to a large region of the genome that appears to be far less compact than is typical of microsporidian genomes, a characteristic which may make this region more amenable to the insertion of foreign genes. The N. locustae catalase gene is expressed in spores, and the protein is detectable by Western blotting. This type of catalase is a particularly robust enzyme that has been shown to function in dormant cells, indicating that the N. locustae catalase may play some functional role in the spore. There is no evidence that the N. locustae catalase functions in a cryptic peroxisome. PMID- 14555491 TI - Developmentally regulated chromosome fragmentation linked to imprecise elimination of repeated sequences in paramecia. AB - The chromosomes of ciliates are fragmented at reproducible sites during the development of the polyploid somatic macronucleus, but the mechanisms involved appear to be quite diverse in different species. In Paramecium aurelia, the process is imprecise and results in de novo telomere addition at locally heterogeneous positions. To search for possible determinants of chromosome fragmentation, we have studied an approximately 21-kb fragmentation region from the germ line genome of P. primaurelia. The mapping and sequencing of alternative macronuclear versions of the region show that two distinct multicopy elements, a minisatellite and a degenerate transposon copy, are eliminated by an imprecise mechanism leading either to chromosome fragmentation and the formation of new telomeres or to the rejoining of flanking sequences. Heterogeneous internal deletions occur between short direct repeats containing TA dinucleotides. The complex rearrangement patterns produced vary slightly among genetically identical cell lines, show non-Mendelian inheritance during sexual reproduction, and can be experimentally modified by transformation of the maternal macronucleus with homologous sequences. These results suggest that chromosome fragmentation in Paramecium is the consequence of imprecise DNA elimination events that are distinct from the precise excision of single-copy internal eliminated sequences and that target multicopy germ line sequences by homology-dependent epigenetic mechanisms. PMID- 14555492 TI - Termite gut symbiotic archaezoa are becoming living metabolic fossils. AB - Over the course of several million years, the eukaryotic gut symbionts of lower termites have become adapted to a cellulolytic environment. Up to now it has been believed that they produce nutriments using their own cellulolytic enzymes for the benefit of their termite host. However, we have now isolated two endoglucanases with similar apparent molecular masses of approximately 36 kDa from the not yet culturable symbiotic Archaezoa living in the hindgut of the most primitive Australian termite, Mastotermes darwiniensis. The N-terminal sequences of these cellulases exhibited significant homology to cellulases of termite origin, which belong to glycosyl hydrolase family 9. The corresponding genes were detected not in the mRNA pool of the flagellates but in the salivary glands of M. darwiniensis. This showed that cellulases isolated from the flagellate cells originated from the termite host. By use of a PCR-based approach, DNAs encoding cellulases belonging to glycosyl hydrolase family 45 were obtained from micromanipulated nuclei of the flagellates Koruga bonita and Deltotrichonympha nana. These results indicated that the intestinal flagellates of M. darwiniensis take up the termite's cellulases from gut contents. K. bonita and D. nana possess at least their own endoglucanase genes, which are still expressed, but without significant enzyme activity in the nutritive vacuole. These findings give the impression that the gut Archaezoa are heading toward a secondary loss of their own endoglucanases and that they use exclusively termite cellulases. PMID- 14555494 TI - Factors influencing the recombinational expansion and spread of telomeric tandem arrays in Kluyveromyces lactis. AB - We have previously shown that DNA circles containing telomeric repeats and a marker gene can promote the recombinational elongation of telomeres in Kluyveromyces lactis by a mechanism proposed to involve rolling-circle DNA synthesis. Wild-type cells acquire a long tandem array at a single telomere, while telomerase deletion (ter1-delta) cells, acquire an array and also spread it to multiple telomeres. In this study, we further examine the factors that affect the formation and spread of telomeric tandem arrays. We show that a telomerase(+) strain with short telomeres and high levels of subtelomeric gene conversion can efficiently form and spread arrays, while a telomere fusion mutant is not efficient at either process. This indicates that an elevated level of gene conversion near telomeres is required for spreading but that growth senescence and a tendency to elongate telomeres in the absence of exogenously added circles are not. Surprisingly, telomeric repeats are frequently deleted from a transforming URA3-telomere circle at or prior to the time of array formation by a mechanism dependent upon the presence of subtelomeric DNA in the circle. We further show that in a ter1-delta strain, long tandem arrays can arise from telomeres initially containing a single-copy insert of the URA3-telomere sequence. However, the reduced rate of array formation in such strains suggests that single-copy inserts are not typical intermediates in arrays formed from URA3 telomere circles. Using heteroduplex circles, we have demonstrated that either strand of a URA3-telomere circle can be utilized to form telomeric tandem arrays. Consistent with this, we demonstrate that 100-nucleotide single-stranded telomeric circles of either strand can promote recombinational telomere elongation. PMID- 14555495 TI - Neutral-lipid analysis reveals elevation of acylglycerols and lack of cholesterol esters in Plasmodium falciparum-infected erythrocytes. AB - Here we show that blood-stage Plasmodium falciparum organisms accumulate a high mass of triacylglycerol and diacylglycerol. However, we failed to detect cholesterol esters, a second neutral lipid species reported to be important for a related apicomplexan, Toxoplasma gondii. Evidence for P. falciparum and T. gondii homologues of acyl coenzyme A:diacylglycerol acyltransferase suggests that acylglycerols may be the conserved neutral lipids in apicomplexans. PMID- 14555493 TI - Posttranslational modifications required for cell surface localization and function of the fungal adhesin Aga1p. AB - Adherence of fungal cells to host substrates and each other affects their access to nutrients, sexual conjugation, and survival in hosts. Adhesins are cell surface proteins that mediate these different cell adhesion interactions. In this study, we examine the in vivo functional requirements for specific posttranslational modifications to these proteins, including glycophosphatidylinositol (GPI) anchor addition and O-linked glycosylation. The processing of some fungal GPI anchors, creating links to cell wall beta-1,6 glucans, is postulated to facilitate postsecretory traffic of proteins to cell wall domains conducive to their functions. By studying the yeast sexual adhesin subunit Aga1p, we found that deletion of its signal sequence for GPI addition eliminated its activity, while deletions of different internal domains had various effects on function. Substitution of the Aga1p GPI signal domain with those of other GPI-anchored proteins, a single transmembrane domain, or a cysteine capable of forming a disulfide all produced functional adhesins. A portion of the cellular pool of Aga1p was determined to be cell wall resident. Aga1p and the alpha-agglutinin Agalpha1p were shown to be under glycosylated in cells lacking the protein mannosyltransferase genes PMT1 and PMT2, with phenotypes manifested only in MATalpha cells for single mutants but in both cell types when both genes are absent. We conclude that posttranslational modifications to Aga1p are necessary for its biogenesis and activity. Our studies also suggest that in addition to GPI-glucan linkages, other cell surface anchorage mechanisms, such as transmembrane domains or disulfides, may be employed by fungal species to localize adhesins. PMID- 14555496 TI - Biosynthesis of glycosylphosphatidylinositol is essential to the survival of the protozoan parasite Toxoplasma gondii. AB - The PIGA gene from Toxoplasma gondii has been cloned and characterized. Like mammalian PIGA, the transmembrane and C-terminal domains are sufficient to direct localization to the parasite endoplasmic reticulum. A functional copy of PIGA is required for tachyzoite viability, demonstrating that glycosylphosphatidylinositol biosynthesis is an essential process in T. gondii. PMID- 14555497 TI - Therapeutic exploitation of the physiological and molecular genetic alterations in head and neck cancer. AB - Despite improvements in the diagnosis and management of head and neck squamous cell carcinomas, there has been minimal increase in the long-term survival in these patients over the last 30 years. Treatment intensification with concurrent chemoradiotherapy has been shown to increase survival and improve organ preservation over radiotherapy alone in patients with locally advanced tumor; however, at a cost of increased long-term toxicity. Recent advances in molecular technology have ushered in a new age of targeted therapy, which holds promise for a better outcome for these patients with potentially less normal tissue toxicity. Some of the new approaches aim to specifically inhibit tumor growth and metastasis by targeting the tumor microenvironment or vasculature, whereas others focus on specific protein or signal transduction pathways. This review will summarize these new molecular and physiological based strategies that can be used for both treatment and chemoprevention of head and neck squamous cell carcinoma. PMID- 14555498 TI - Natural T cell immunity against cancer. AB - It has long been a matter of debate whether tumors are spontaneously immunogenic in patients. With the availability of sensitive methods, naturally occurring T cells directed against tumor-associated antigens (TAAs) can be frequently detected in cancer patients. In this review, we summarize the current data on T cell responses to TAAs in various malignancies, including melanoma, colorectal cancer, leukemia, and breast cancer. T cell responses against various antigens, including melanoma differentiation antigens, carcinoembryonic antigen, epithelial cell adhesion molecule, her-2/neu, Wilms' tumor protein, proteinase 3, NY-ESO-1, and surviving, have been reported in a substantial number of patients. In contrast, other TAAs, including most antigens of the MAGE family, do not usually elicit spontaneous T cell responses. A distinction between direct ex vivo T cell responses and in vitro-generated T cell responses is provided because in vitro stimulation results in quantitative and functional changes of T cell responses. The possible role of TAA-specific T cells in immunosurveillance and tumor escape and the implications for immunological treatment strategies are discussed. Naturally occurring T cells against TAAs are a common phenomenon in tumor patients. Understanding the mechanisms and behavior of natural TAA-specific T cells could provide crucial information for rational development of more efficient T cell-directed immunotherapy. PMID- 14555499 TI - Ribonucleotide reductase subunit one as gene therapy target: commentary re: M-Y. Cao et al., Adenovirus-mediated ribonucleotide reductase R1 gene therapy of human colon adenocarcinoma. Clin. Cancer Res., 9: 4304-4308, 2003. PMID- 14555500 TI - Fulvestrant in postmenopausal women with advanced breast cancer. AB - PURPOSE: Patients with hormone-sensitive breast cancer who have responded to tamoxifen (TAM) may receive additional benefit from a second endocrine agent after progression or relapse after TAM therapy. Fulvestrant (FVT; Faslodex; i.m. injection, ICI 182,780; AstraZeneca Pharmaceuticals, Wilmington, DE) was developed as a selective antagonist of estrogen. In postmenopausal women, FVT is reported to inhibit the proliferative effects of estrogen on sensitive tissues and has no apparent measurable estrogenic activity. In this report, we describe the data and analyses supporting marketing approval for FVT by the United States Food and Drug Administration (FDA). EXPERIMENTAL DESIGN: The FDA review of 16 clinical trials and 6 pharmacokinetic trials, as well as preclinical pharmacology and chemistry data, are described. The bases for marketing approval are summarized. RESULTS: Toxicology studies in the mouse, rat, and dog showed minimal toxicity except for antiestrogenic effects. Because of FVT aqueous insolubility, an i.m. formulation, given at monthly intervals, was selected for clinical studies. Pharmacokinetic studies demonstrated sustained concentrations with monthly injection. In in vitro studies FVT was extensively metabolized, primarily by hepatic cytochrome P450 3A4. Phase I studies showed minimal toxicity, and the maximal dose (250 mg) was limited by FVT solubility. In two Phase III trials, 851 patients were randomized to either 250 mg FVT i.m. monthly or to anastrozole (ANZ) 1 mg p.o. daily. Ninety-six percent of patients had received TAM previously for early (adjuvant treatment) or advanced breast cancer. Response rates (RR) were 17% for both FVT and ANZ study arms in the North American trial, and were 20% versus 15% for FVT versus ANZ, respectively, in the European trial. There were no observed differences between study arms with respect to time to progression or survival. The most common FVT adverse events reported as potentially treatment-related were injection site reactions and hot flashes. CONCLUSIONS: FVT was approved on April 25, 2002 by the FDA for the treatment of hormone receptor-positive metastatic breast cancer in postmenopausal women with disease progression after antiestrogen therapy. The recommended dose is 250 mg i.m. monthly as a single 5 ml injection or as two concurrent 2.5 ml injections into the buttocks. Approval was based on results of two randomized trials comparing response rates and time to progression of FVT- and ANZ-treated patients. Complete prescribing information is available on the FDA website. PMID- 14555501 TI - Variability in response assessment in solid tumors: effect of number of lesions chosen for measurement. AB - PURPOSE: This study was performed to systematically evaluate the variability in tumor response assessments that occurs depending on how many tumor deposits are selected for measurement at imaging. EXPERIMENTAL DESIGN: The two largest perpendicular diameters of all tumor deposits in 36 patients were measured on computed tomography scans obtained at baseline and first posttherapy follow-up. A computerized modeling analysis of those data was performed to determine each patient's therapeutic response category assignment for every possible number of lesions in a grouping. The variance in the sum of measurements of these lesion groupings was calculated, and the frequency of response assessment categories was plotted against the number of lesions. RESULTS: The computerized analysis of the resultant 1,833,821 possible combinations of tumor deposits showed that when six lesions were measured bidimensionally and four lesions were measured undimensionally, the average variance decreased by 90%. The number of different response assessment categories into which a patient was assigned decreased with increasing lesion grouping size. When six or more lesions were measured bidimensionally, 9% of all possible lesion groupings still fell into a second response category, reflecting the effect of which particular lesions are chosen for measurement. CONCLUSIONS: Measuring larger numbers of lesions will decrease the variance. In this population, the variance decreased by at least 90% when six or more lesions were measured bidimensionally. Further confirmatory studies with larger series of patients are warranted before adopting this number as a criterion in clinical trials for assessing the activity of antineoplastic therapies. PMID- 14555503 TI - The relationship of human wound vascular endothelial growth factor (VEGF) after breast cancer surgery to circulating VEGF and angiogenesis. AB - PURPOSE: The assessment of locally produced proangiogenic cytokines may be an indicator of the stromal response of an individual to wounding or cancer. This study describes the profile of VEGF production in human surgical wounds in both breast cancer patients and reduction mammoplasty controls, and assesses the changes in systemic VEGF levels and platelet profiles perioperatively. EXPERIMENTAL DESIGN: Perioperative surgical wound fluid samples and blood were collected daily up to 13 days from 52 patients undergoing breast cancer surgery (local tumor burden), delayed breast reconstruction (previous tumor burden but none present at the time of surgery), or breast reduction surgery (noncancer control). Samples were analyzed for VEGF by ELISA RESULTS: VEGF levels in surgical wound fluid were lowest on day 1 followed by an early peak on day 2 of >900% the corresponding serum value. There was a trend in the VEGF response at the day-2 time point: reduction > reconstruction > cancer subgroups, with a significant difference between the reduction and cancer subgroups (P < 0.05). There was a 20-30-fold variation in the response between days 1 and 2, and within subgroups. CONCLUSIONS: Much higher local concentrations of angiogenic factors may need to be antagonized for effective antiangiogenic therapy, and there is great heterogeneity between patients. The small peripheral blood changes compared with large tumor fluid changes show that there is a tissue barrier. This has relevance for design of antiangiogenic therapy trials, highlighting the need for individually tailored treatment with biologically targeted interventions. PMID- 14555502 TI - Celecoxib modulates the expression of cyclooxygenase-2, ki67, apoptosis-related marker, and microvessel density in human cervical cancer: a pilot study. AB - PURPOSE: We investigated whether a short treatment with the cyclooxygenase-2 (COX 2) inhibitor celecoxib could modulate Ki67 antigen and the caspase cleavage product of keratin 18, recognized as a marker of early apoptosis. The activity of celecoxib on microvessel density (MVD) and angio-power Doppler sonography-derived indices of tumor vascularization was also assessed. Serum levels of squamous cell carcinoma antigen and the proliferative potential and subsets of peripheral T cells before and after celecoxib treatment were also analyzed. EXPERIMENTAL DESIGN: Tumor biopsy specimens from 14 patients with cervical cancer were obtained at baseline and after 10 days of celecoxib treatment (400 mg twice daily). Tumor and stroma COX-2 expression, Ki67, apoptosis, and MVD were assessed by immunohistochemistry, whereas prostaglandin E(2) levels were measured by RIA. RESULTS: At baseline, COX-2 integrated density values in tumor compartment ranged from 10.7 to 60.1 (median, 26.5) and were significantly higher than tumor COX-2 integrated density values after celecoxib treatment (range, 0.6-42.3; median, 12.6; P = 0.0043). The percentages of Ki67-positive tumor cells in pre-celecoxib cases ranged from 39.3 to 87.4 (median, 50.8) and were significantly higher than the percentage in the corresponding posttreatment samples (range, 27.7-83.8; median, 43.1; P = 0.0092). MVD values in pre-celecoxib biopsies ranged from 28.0 to 55.0 (median, 38.5) and were significantly higher than the corresponding values in posttreatment samples (range, 16.0-49.5; median; 27.6; P = 0.012). Also, prostaglandin E(2) levels showed a trend to be reduced after celecoxib treatment (range: 4.7-386.6 pg/mg wet tissue in pretreated cases versus 4.8-91.9 pg/mg wet tissue in posttreated cases (P = 0.092). CONCLUSIONS: In cervical cancer, celecoxib treatment decreases tumor COX-2 expression and markers of proliferation and neoangiogenesis, while being uneffective on stroma COX-2 levels, thus suggesting that selective COX-2 inhibitors may be a promising strategy not only for chemopreventive approaches but also for therapeutic approaches in this neoplasia. PMID- 14555504 TI - Resistance to gefitinib in PTEN-null HER-overexpressing tumor cells can be overcome through restoration of PTEN function or pharmacologic modulation of constitutive phosphatidylinositol 3'-kinase/Akt pathway signaling. AB - PURPOSE: Tyrosine kinase (TK) inhibitors are emerging as a promising new approach to the treatment of HER overexpressing tumors, however optimal use of these agents awaits further definition of the downstream signaling pathways that mediate their effects. We reported previously that both EGFR- and Her2 overexpressing tumors are sensitive to the new EGFR-selective TK inhibitor gefitinib (ZD1839, "Iressa"), and sensitivity to this agent correlated with its ability to down-regulate Akt. However, EGFR-overexpressing MDA-468 cells, which lack PTEN function, are resistant to ZD1839, and ZD1839 is unable to down regulate Akt activity in these cells. EXPERIMENTAL DESIGN: To study the role of PTEN function, we generated MDA468 cells with tet-inducible PTEN expression. RESULTS: We show here that the resistance of MDA-468 cells to ZD1839 is attributable to EGFR-independent constitutive Akt activation caused by loss of PTEN function in these cells. Reconstitution of PTEN function through tet inducible expression restores ZD1839 sensitivity to these cells and reestablishes EGFR-stimulated Akt signaling. Although restoration of PTEN function to tumors is difficult to implement clinically, much of the effects of PTEN loss are attributable to overactive PI3K/Akt pathway signaling, and this overactivity can be modulated by pharmacologic approaches. We show here that pharmacologic down regulation of constitutive PI3K/Akt pathway signaling using the PI3K inhibitor LY294002 similarly restores EGFR-stimulated Akt signaling and sensitizes MDA-468 cells to ZD1839. CONCLUSIONS: Sensitivity to ZD1839 requires intact growth factor receptor-stimulated Akt signaling activity. PTEN loss leads to uncoupling of this signaling pathway and results in ZD1839 resistance, which can be reversed with reintroduction of PTEN or pharmacologic down-regulation of constitutive PI3K/Akt pathway activity. These data have important predictive and therapeutic clinical implications. PMID- 14555505 TI - Amplification of virus-induced antimelanoma T-cell reactivity by high-dose interferon-alpha2b: implications for cancer vaccines. AB - PURPOSE: The therapeutic effectiveness of cancer vaccines, composed of tumor antigens that are also self-antigens, may be limited by the normal mechanisms that preserve immunological tolerance. Consistent with this notion, we found that vaccination of melanoma patients with recombinant viral vaccines expressing gp100 (a melanoma antigen also expressed by normal melanocytes) produced only transient increases in noncytotoxic T cells specific for immunodominant gp100 epitopes. To improve the therapeutic effects of these vaccines, IFN-alpha2b (IFN-alpha) was administered to some high-risk patients. EXPERIMENTAL DESIGN: 7 HLA-A*0201(+) patients were injected with high doses of IFN-alpha (20 MU/m(2) x 20 doses) at various times after completing the vaccination protocol. Clinical toxicity and responses were documented, and the effects on gp100-reactive T cells were measured by IFN-gamma enzyme-linked immunospot assays, tetramers of HLA-A*0201 and gp100 epitopes, and cellular cytotoxicity assays. RESULTS: In patients who had previously responded to vaccination, high doses of IFN-alpha recalled gp100 reactive T cells with the ability to kill gp100-expressing tumor targets in vitro. Concomitant with the reappearance of these cytotoxic T cells, tumor regression was observed in the two patients with clinically evident metastatic disease. CONCLUSIONS: The finding that high-dose IFN recalls previously activated tumor-reactive T cells with potent killing ability suggests a strategy to maintain antitumor responses initiated by cancer vaccines. PMID- 14555506 TI - Tirapazamine plus carboplatin and paclitaxel in advanced malignant solid tumors: a california cancer consortium phase I and molecular correlative study. AB - PURPOSE: Tumor hypoxia confers chemotherapy resistance. Tirapazamine is a cytotoxin that selectively targets hypoxic cells and has supra-additive toxicity with platinums and taxanes in preclinical studies. We conducted a Phase I study of tirapazamine, carboplatin, and paclitaxel and assessed potential plasma markers of hypoxia as surrogates for response. EXPERIMENTAL DESIGN: Forty-two patients with advanced solid tumors were treated at four dose levels; parallel dose escalations were carried out in chemotherapy-naive and previously treated subjects. Pre and post-therapy plasma levels of the hypoxia-induced proteins plasminogen activator inhibitor-1 and vascular endothelial growth factor were measured. RESULTS: Three of four chemotherapy-naive patients developed dose limiting toxicities at dose level 4 (grade 3 stomatitis/infection, grade 3 emesis, and grade 4 febrile neutropenia). Four of seven previously treated patients developed dose-limiting toxicities at dose level 3, including one death [grade 3 myalgia, grade 3 infection/grade 4 neutropenia, grade 3 infection/grade 4 neutropenia, and grade 5 infection (death)/grade 4 neutropenia]. Of 38 patients assessable for response, 3 had a complete response, 1 a partial response, 1 an unconfirmed partial response, and 23 had stable disease in at least one evaluation; 10 quickly progressed. One complete responder had normalization of vascular endothelial growth factor and plasminogen activator inhibitor-1 levels. CONCLUSION: Dose levels 3 (carboplatin AUC of 6, 225 mg/m(2) paclitaxel, and 330 mg/m(2) tirapazamine) and 2 (carboplatin AUC 6, 225 mg/m(2) paclitaxel, and 260 mg/m(2) tirapazamine) are the maximum tolerated doses for chemotherapy naive and patients treated previously, respectively. Dose level 3 is the experimental arm of a Phase III Southwest Oncology Group trial (S0003) in advanced non-small cell lung cancer. Potential markers of tumor hypoxia may be useful correlates in studies of hypoxic cytotoxins and are being prospectively investigated in S0003. PMID- 14555507 TI - Dihydropyrimidinase deficiency and severe 5-fluorouracil toxicity. AB - Dihydropyrimidinase (DHP) is the second enzyme in the catabolism of 5 fluorouracil (5FU), and it has been suggested that patients with a deficiency of this enzyme are at risk from developing severe 5FU-associated toxicity. In this study, we demonstrated for the first time that in one patient the severe toxicity, after a treatment with 5FU, was attributable to a partial deficiency of DHP. Analysis of the DHP gene showed that the patient was heterozygous for the missense mutation 833G>A (G278D) in exon 5. Heterologous expression of the mutant enzyme in Escherichia coli showed that the G278D mutation leads to a mutant DHP enzyme without residual activity. An analysis for the presence of this mutation in 96 unrelated Dutch Caucasians indicates that the allele frequency in the normal population is <0.5%. Our results show that a partial DHP deficiency is a novel pharmacogenetic disorder associated with severe 5FU toxicity. PMID- 14555508 TI - Reduced MLH1 expression after chemotherapy is an indicator for poor prognosis in esophageal cancers. AB - PURPOSE: Loss of function or expression of the mismatch repair gene MLH1 has been implicated in experimentally acquired resistance to cisplatin (CDDP) and other anticancer agents. The clinical significance of MLH1 expression was evaluated in advanced thoracic squamous cell carcinoma of the esophagus (ESCC) treated by neoadjuvant chemotherapy. EXPERIMENTAL DESIGN: We investigated MLH1 and P53 expression by immunohistochemistry in the surgical specimens of 107 patients who had undergone preoperative chemotherapy using CDDP along with 5-FU and ADM. These findings were correlated with the clinical outcome for this treatment. Biopsy samples before chemotherapy in 20 of these patients, and another 43 surgical specimens without chemotherapy, were also examined as control samples. RESULTS: In surgical specimens of ESCC, low MLH1 expression was not frequent without chemotherapy, whereas it was commonly observed after chemotherapy (14 versus 37%, P = 0.0057). Comparison between samples before and after chemotherapy revealed that MLH1 expression was unchanged during chemotherapy in 12 of 20 patients (60%) but was from high to low in 8 of 20 patients (40%). In the surgical specimen after neoadjuvant chemotherapy, MLH1 expression was not correlated with any clinicopathological factors, including the response to chemotherapy. However, low MLH1 showed poorer prognosis than high MLH1 (5-year survival 40.6 versus 19.3%, P = 0.0393), and in multivariate analysis, MLH1 was an independent prognostic factor for this multimodal treatment, following lymph node metastasis and clinical response to chemotherapy. Positive p53 expression, which was not affected by chemotherapy, was weakly associated with a poor response and clinical outcome, although this trend was not significant. CONCLUSIONS: In advanced ESCC, expression of MLH1 is reduced during CDDP-based chemotherapy, and this may partly account for poor postoperative survival. PMID- 14555509 TI - Evaluation of pre-existent immunity in patients with primary breast cancer: molecular and cellular assays to quantify antigen-specific T lymphocytes in peripheral blood mononuclear cells. AB - PURPOSE: Breast cancers are known to frequently (over)express several well characterized tumor-associated antigens (TAAs) such as carcinoembryonic antigen, MUC-1, Her-2/neu, and cancer/testis antigens such as NY-ESO-1, SSX-2, and members of the MAGE family. Whereas in melanoma patients, the detection of pre-existing T cell responses to tumor-associated differentiation antigens was a rationale to initiate several vaccination strategies, little is known thus far concerning tumor-specific immunity in breast cancer patients. The objectives of our study were (a) to modify and compare different immunodiagnostic T cell assays with regard to their suitability for clinical applications and (b) to determine endogenous TAA-specific T cell immunity of breast cancer patients at the time point of primary diagnosis. EXPERIMENTAL DESIGN: Using MUC-1- and Her-2/neu derived HLA-A*0201-restricted peptides as model antigens, we analyzed antigen dependent IFN-gamma release of T cells by enzyme-linked immunospot (ELISpot) assay, intracellular cytokine flow cytometry (CytoSpot), and quantitative real time PCR. As an assay independent of T cell function, we performed tetramer staining. RESULTS: In our hands, the quantitative real-time PCR method is most sensitive and a feasible screening test to perform an "immunological staging" of cancer patients. By doing this, we detected in 7 of 13 (54%) of HLA-A*0201(+) breast cancer patients a pre-existent specific cellular immune response to at least one of the investigated TAAs (MUC-1, Her-2/neu, carcinoembryonic antigen, NY-ESO-1, and SSX-2). Four of 21 patients (19%) were found to have a significant Her-2/neu-specific T cell response as defined by a stimulation index >/==" BORDER="0"> 2 (range, 10-88). CONCLUSIONS: Although the clinical relevance of endogenous TAA-specific immunity remains unclear, our findings suggest that patients with primary breast cancer can mount a T cell immune response to their tumor that might be beneficially enhanced by TAA-dependent vaccination strategies in the adjuvant situation. PMID- 14555510 TI - Pretreatment proliferation parameters do not add predictive power to clinical factors in cervical cancer treated with definitive radiation therapy. AB - PURPOSE: To examine the prognostic value of tumor proliferation measurements in women with carcinoma of the uterine cervix. We report an update of a prospective study focusing on whether pretreatment proliferation parameters are associated with clinical outcome, relative to other established clinical factors. MATERIALS AND METHODS: One hundred and one patients were recruited into the study from years 1991 to 1999. The LI for in vivo bromodeoxyuridine incorporation by the tumor and the potential doubling time (T(pot)) were determined by flow cytometry (fc). LI and its staining pattern were also assessed by immunohistochemistry (ih) using tissue sections. Apoptosis was assessed histologically using morphological criteria. Patients were treated with definitive radiation therapy. RESULTS: A successful fc measurement for LI-fc and T(pot) was possible in 95 patients (94%). The median/mean LI-fc was 6.6/7.6% (range 1.4-36.1%), and for LI-ih, 10.8/11.5%. To date, 43 patients have died of disease, and the median follow-up for alive patients is 6.2 years (range 1.3-9.3 years). Among 88 patients who completely responded to treatment, 40 patients have relapsed (14 pelvic, 23 distant, and 3 pelvic and distant). In univariate analysis, the significant factors for adverse disease-free survival were large tumor size (P = 0.0001), low hemoglobin (P = 0.001), pelvic lymph node status (P = 0.004), stage (P = 0.013), and overall treatment time (P = 0.0008). In multivariate analysis, only tumor size, pelvic lymph node status, and overall treatment time remained significant for disease free survival. LI-fc, LI-ih, T(pot), ploidy, pattern of bromodeoxyuridine staining, and apoptosis were not significantly associated with clinical outcome in univariate or multivariate analyses. CONCLUSIONS: These mature data indicate that none of the pretreatment proliferation parameters have prognostic significance in the radical radiotherapy of carcinoma of the uterine cervix, despite the significance of overall treatment time for treatment outcome. PMID- 14555511 TI - Polymorphisms in BRCA1 and BRCA2 and risk of epithelial ovarian cancer. AB - PURPOSE: Because inherited BRCA1 or BRCA2 mutations strikingly increase ovarian cancer risk, polymorphisms in these genes could represent low penetrance susceptibility alleles. Previous studies of the BRCA2 N372H polymorphism suggested that HH homozygotes have a modestly increased risk of both breast and ovarian cancer. We have examined whether BRCA2 N372H or common amino acid changing polymorphisms in BRCA1 predispose to ovarian cancer. EXPERIMENTAL DESIGN: A population-based, case control study of ovarian cancer was performed in North Carolina. Cases included 312 women with ovarian cancer (76% invasive and 24% borderline) and 401 age- and race-matched controls. Blood DNA from subjects was genotyped for BRCA2 N372H and BRCA1 Q356R and P871L. RESULTS: There was no association between BRCA2 N372H and risk of borderline or invasive epithelial ovarian cancer. The overall odds ratio (OR) for HH homozygotes was 0.8 [95% confidence interval (CI) = 0.4-1.5] and was similar in all subsets, including invasive serous cases. In addition, neither the BRCA1 Q356R (OR = 0.9, 95% CI 0.5 1.4) nor P871L (OR = 0.9, 95% CI 0.6-1.9) polymorphisms were associated with ovarian cancer risk. There was a significant racial difference in allele frequencies of the P871L polymorphism (P = 0.64 in Caucasians, L = 0.76 in African-Americans, P < 0.0001). CONCLUSIONS: In this population-based, case control study, common amino acid changing BRCA1 and 2 polymorphisms were not found to affect the risk of developing ovarian cancer. PMID- 14555512 TI - Increased populations of regulatory T cells in peripheral blood and tumor infiltrating lymphocytes in patients with gastric and esophageal cancers. AB - PURPOSE: It is well known that tumor-infiltrating lymphocytes (TILs) and, to a lesser extent, peripheral blood lymphocytes from patients with advanced-stage cancer have a poor immune response. Regulatory T cells (T-regs), characterized by coexpression of CD4 and CD25 markers, can inhibit the immune response mediated by CD4+/CD25- and CD8+ T cells. In the present study, we evaluated the prevalence of T-regs in peripheral blood and TILs in patients with gastric and esophageal cancers. EXPERIMENTAL DESIGN: The population of CD4+/CD25+ cells as a percentage of total CD3+ cells was evaluated by flow cytometric analysis with triple-color staining. To assess the functional activity of CD4+/CD25+ cells, CD4+/CD25+ or CD4+/CD25- cells were purified from peripheral blood mononuclear cells with magnetic beads. The cytokine production [interleukin (IL)-10 and IFN-gamma] from the CD4+/CD25+ cells in response to anti-CD3 stimulation was evaluated. Also, the antiproliferative function of CD4+/CD25+ cells was measured by evaluating the proliferative activity of CD4+/CD25- cells in response to anti-CD3 plus anti-CD28 in the presence of autologous CD4+/CD25+ cells. RESULTS: The prevalence of peripheral blood CD4+/CD25+ cells in both gastric (n = 20; 14.2 +/- 4.9%) and esophageal cancer patients (n = 10; 19.8 +/- 6.9%) was significantly higher than that in healthy donors (n = 16; 7.2 +/- 2.1%). The population of CD4+/CD25+ cells in the TILs of gastric cancer patients with advanced disease (19.8 +/- 4.5%) was significantly higher than that in TILs of patients with early-stage disease (4.8 +/- 2.1%) or that in intraepithelial lymphocytes of normal gastric mucosa (4.0 +/ 1.2%). As a functional consequence, CD4+/CD25+ cells did not produce IFN-gamma, whereas CD4+/CD25- cells secreted IFN-gamma. Moreover, CD4+/CD25+ cells produced large amounts of IL-10, whereas CD4+/CD25- cells secreted little IL-10. The proliferation of CD4+/CD25- cells was inhibited in the presence of CD4+/CD25+ cells in a dose-dependent manner, confirming that CD4+/CD25+ has an inhibitory activity corresponding to T-regs. CONCLUSIONS: The populations of CD4+/CD25+ T regs in peripheral blood and TILs in patients with gastric and esophageal cancers were significantly higher in comparison with those in healthy donors or normal mucosa. PMID- 14555513 TI - Increased expression of integrin-linked kinase is correlated with melanoma progression and poor patient survival. AB - PURPOSE: Integrin-linked kinase (ILK), a key component of the extracellular matrix adhesion, has been studied extensively in recent years. Overexpression of ILK in epithelial cells results in anchorage-independent cell growth with increased cell cycle progression. Furthermore, increased ILK expression is correlated with progression of several human tumor types, including breast, prostate, and colon carcinomas. However, the role of ILK overexpression in human melanoma pathogenesis is not known. To investigate whether ILK plays a role in melanoma progression, we measured ILK expression in primary melanoma biopsies at various stages of invasion and evaluated the prognostic value of ILK expression in human melanoma. EXPERIMENTAL DESIGN: We used tissue microarray and immunohistochemistry to determine ILK expression in 67 primary melanomas and analyzed the correlation between ILK expression and melanoma progression and 5 year patient survival. RESULTS: We show that strong ILK expression is significantly associated with melanoma thickness. Strong ILK expression was observed in 0, 22, 33, and 63% in melanoma biopsies 3.0 mm in thickness, respectively. Furthermore, strong ILK expression was detected in 83% of the tumors with lymph node invasion compared with only 18% for tumors without lymph node invasion (P < 0.01). Strikingly, our data revealed that strong ILK expression is inversely correlated with 5-year patient survival (P < 0.05). CONCLUSION: ILK expression increases dramatically with melanoma invasion and progression and is inversely correlated with patient survival. PMID- 14555514 TI - Expression analysis of DNA methyltransferases 1, 3A, and 3B in sporadic breast carcinomas. AB - PURPOSE: Three genes, namely DNA methyltransferase (DNMT) 1, DNMT3A, and DNMT3B, coding for DNMTs that affect promoter methylation status are thought to play an important role in the development of cancers. Little is known of the biological and clinical significance of these genes in human breast cancer. EXPERIMENTAL DESIGN: We used real-time reverse transcription-PCR assays to quantify the mRNA expression of the three DNMT genes in a series of 130 breast cancer patients. We also sought relationships between mRNA levels of the DNMTs and those of 20 target genes involved in the DNMT pathway (subgroup of 46 breast tumors). RESULTS: The DNMT3B gene showed the highest range of expression (81.8 compared with 16.6 and 14 for DNMT1 and DNMT3A, respectively). DNMT3B was overexpressed in 30% of the patients (5.4 and 3.1% for DNMT1 and DNMT3A, respectively). DNMT3B overexpression was significantly related to Scarff, Bloom, and Richardson histopathological grade III (P = 0.002), ERalpha negativity (P = 0.0015), and strong MKI67 expression (P = 3 x 10(-6)). In univariate analysis, DNMT3B overexpression was associated with poor relapse-free survival in the subgroup of patients who received adjuvant hormone therapy (with or without chemotherapy; P = 0.0064). Although the poor prognosis associated with DNMT3B overexpression was confirmed by univariate analysis in an independent series of 98 postmenopausal women exclusively treated with adjuvant tamoxifen therapy (P = 0.0036), DNMT3B expression status did not persist as an independent prognostic factor in multivariate analysis. CONCLUSIONS: Although we failed to identify underexpression of specific target genes associated with DNMT increasing expression, the frequent overexpression of DNMT3B in this breast tumor series points to DNMT3B as a potential new therapeutic target in breast cancer. PMID- 14555515 TI - High levels of serum HER-2/neu and YKL-40 independently reflect aggressiveness of metastatic breast cancer. AB - PURPOSE: To evaluate serum levels of HER2 (an epithelial growth factor) and YKL 40 (a growth factor participating in inflammation and remodeling of the extracellular matrix) in relation to outcome in patients with their first diagnosis of recurrent breast cancer. DESIGN: Serum HER2 and YKL-40 levels were measured in 100 patients referred with their first metastatic manifestation of breast cancer before first line anthracycline-based therapy and related to response to therapy, metastatic pattern, time to progression, and overall survival. During the observation period of 64-84 months, 89 patients died of breast cancer. RESULTS: The patients had higher serum HER2 and YKL-40 levels than healthy females (P < 0.0001). Serum HER2 was elevated in 32% of the patients and serum YKL-40 in 30%. These patients were more sick (P < 0.01) and more often had parenchymal involvement (P < 0.0005), especially liver metastases (P < 0.00005). In multivariate Cox analysis, high serum levels of HER2 or YKL-40 or lack of estrogen receptors independently doubled the relative risk of progression and dying (P < 0.001) even after accounting for other independent prognostic variables, such as axillary nodal involvement at primary diagnosis, liver metastases, and more than two metastatic sites. Fewer patients with high serum HER2 or YKL-40 or lack of estrogen receptors responded with a complete remission on chemotherapy (P = 0.005, 0.036, and 0.006). In these patients, high serum YKL 40 was a stronger predictor of survival than high serum HER2 or lack of estrogen receptors. CONCLUSIONS: High serum HER2 and YKL-40 independently identified subgroups of patients with metastatic breast cancer with a poor prognosis. PMID- 14555516 TI - Meningiomas: analysis of loss of heterozygosity on chromosome 10 in tumor progression and the delineation of four regions of chromosomal deletion in common with other cancers. AB - PURPOSE: Loss of heterozygosity (LOH) of alleles on chromosome 10 has been reported in many cancers, leading to the identification of tumor suppressor genes on this chromosome. Several reports implicate LOH of chromosome 10 alleles in meningioma progression, but the frequency and complexity of the loss have not been well characterized. Furthermore, the location and identity of the putative tumor suppressor genes on this chromosome that contribute to meningioma progression are unknown because the currently characterized tumor suppressor genes do not appear to be involved. Therefore, this study was undertaken to (a) assess the frequency and complexity of LOH in meningioma progression, (b) map the LOH patterns of individual meningiomas to define the smallest regions of shared chromosomal deletion, and (c) compare the identified regions with chromosome 10 deletions in other cancers, and thereby initiate the localization of the putative tumor suppressor genes. EXPERIMENTAL DESIGN: We examined 11 microsatellite dinucleotide repeat loci in 208 meningiomas of all grades using laser capture microdissection and fluorescence-based detection of PCR products. RESULTS: For all markers examined, the incidence of LOH was much higher in all grades than that previously reported, with incidence and complexity of LOH increasing with tumor grade. LOH mapping identified four regions of chromosomal deletion: 10pter D10S89, D10S109-D10S215, D10S187-D10S209, and D10S169-10qter. These deletions on chromosome 10 are shared with other cancer types. CONCLUSIONS: These results delineate chromosomal locations of putative tumor suppressor genes on chromosome 10 that likely play an early role in meningioma tumorigenesis as well as tumor progression. PMID- 14555517 TI - Meningiomas: loss of heterozygosity on chromosome 10 and marker-specific correlations with grade, recurrence, and survival. AB - PURPOSE: In a study of 208 meningiomas, we found a high incidence of loss of heterozygosity (LOH) on chromosome 10 in benign (73.4%), atypical (80.0%), and malignant (86.7%) tumors. A large percentage of the benign and atypical tumors and an increasing percentage of malignant tumors had LOH on multiple loci (43.9%, 45%, and 66.7%, respectively). The high incidence of LOH occurring early in meningioma progression suggests that LOH at individual alleles may serve as a marker of clinically relevant alterations useful for patient diagnosis, the subclassification of tumors, and/or the treatment of patients. EXPERIMENTAL DESIGN: To test this, we examined 208 sporadic and recurrent meningiomas of all grades for correlations between LOH at 11 markers on chromosome 10 and tumor location, histology, and grade and patient race, gender, age, recurrence, and survival. RESULTS: Several significant correlations were found. The data indicate that genetic differences occur not only between tumors of different grade, but also between tumors of the same grade, and therefore may be useful to define genetic subsets with clinical implications. LOH at D10S179 (P = 0.001) or D10S169 (P = 0.004) is most likely present in higher-grade meningiomas and, when present in benign tumors, may signify sampling error or a morphologically benign but biologically aggressive tumor. Furthermore, LOH at D10S209 (P = 0.06) and D10S169 (P = 0.01) may predict shorter survival and/or higher rates of recurrence, respectively, in tumors with benign or malignant histology. CONCLUSIONS: We conclude that these chromosome 10 markers deserve further testing as unfavorable prognostic indicators for meningioma patients. PMID- 14555518 TI - BRCA2 mutations and androgen receptor expression as independent predictors of outcome of male breast cancer patients. AB - PURPOSE: Germline mutations of the BRCA2 gene are involved in the development of a considerable number of male breast cancer cases. Although phenotypic differences have been observed between sporadic and BRCA-related breast carcinomas, conflicting data exist on the differences in prognosis of women with hereditary and sporadic breast cancer. The purpose of the study was to investigate the prognostic value of BRCA2 status in male breast carcinoma (MBC). EXPERIMENTAL DESIGN: We studied 43 male breast cancer patients, including 12 with BRCA2 mutations. Tumor samples were characterized immunohistochemically using antibodies to estrogen receptor, progesterone receptor, and androgen receptor (AR). RESULTS: BRCA2-related tumors presented at the earlier age compared with sporadic tumors (P = 0.005). Patients positive and negative for BRCA2 mutations did not differ with respect to tumor size, lymph node involvement, histological grade, and sex hormone receptor status. Five-year disease-free survival (DFS) and overall survival (OS) were significantly decreased in BRCA2-positive patients (67% versus 28% for BRCA2-negative versus positive patients, respectively, P = 0.017 for DFS; 86% versus 25%, P = 0.006 for OS). Shorter survival was also correlated with expression of AR in tumor tissue (74% versus 33% for patients with tumors staining negatively and positively for AR, P = 0.029 for DFS; 71% versus 57%, P = 0.05 for OS). CONCLUSIONS: The BRCA2 mutations and AR expression in tumor tissue are independent adverse factors for MBC prognosis. BRCA2-related MBC presents at the earlier age compared with non-BRCA2-related cancer, but do not differ with respect to other clinicopathological features. PMID- 14555519 TI - HLA-G is a potential tumor marker in malignant ascites. AB - PURPOSE: Molecular approaches as supplements to cytological examination of malignant ascites may play an important role in the clinical management of cancer patients. HLA-G is a potential tumor-associated marker and that one of its isoforms, HLA-G5, produces a secretory protein. This study is to assess the clinical utility of secreted HLA-G levels in differential diagnosis of malignant ascites. EXPERIMENTAL DESIGN: We used ELISA to assess whether secretory HLA-G (sHLA-G) could serve as a marker of malignant ascites in ovarian and breast carcinomas, which represent the most common malignant tumors causing ascites in women. RESULTS: On the basis of immunohistochemistry, 45 (61%) of 74 ovarian serous carcinomas and 22 (25%) invasive ductal carcinomas of the breast demonstrated HLA-G immunoreactivity ranging from 2 to 100% of the tumor cells. HLA-G staining was not detected in a wide variety of normal tissues, including ovarian surface epithelium and normal breast tissue. Revese transcription-PCR demonstrated the presence of HLA-G5 isoform in all of the tumor samples expressing HLA-G. ELISA was performed to measure the sHLA-G in 42 malignant and 18 benign ascites supernatants. sHLA-G levels were significantly higher in malignant ascites than in benign controls (P < 0.001). We found that the area under the receiver-operating characteristic curve for sHLA-G was 0.95 for malignant versus benign ascites specimens. At 100% specificity, the highest sensitivity to detect malignant ascites was 78% (95% confidence interval, 68-88%) at a cutoff of 13 ng/ml. CONCLUSIONS: Our findings suggest that measurement of sHLA-G is a useful molecular adjunct to cytology in the differential diagnosis of malignant versus benign ascites. PMID- 14555520 TI - Gene-specific formation and repair of DNA monoadducts and interstrand cross-links after therapeutic exposure to nitrogen mustards. AB - PURPOSE: To investigate the possibility of measuring the gene-specific DNA damage after therapeutic exposure to nitrogen mustards and to examine its relationship with the clinical response. EXPERIMENTAL DESIGN: The kinetics of gene-specific monoadducts and interstrand cross-link formation/repair were measured in the p53 and N-ras genes. DNA extracted from human peripheral lymphocytes following in vitro exposure to melphalan or therapeutic exposure to melphalan or cyclophosphamide was used. RESULTS: When lymphocytes were treated in vitro with biologically relevant doses of melphalan, monoadducts accumulated rapidly in both p53 and N-ras genes, reaching maximal levels within 2 h, whereas the highest interstrand cross-link levels were found within 8 h. Thereafter, the adducts were repaired with half-lives of 14.5 +/- 0.3 h (p53) or 18.8 +/- 1.5 h (N-ras) for monoadducts and 12.4 +/- 0.8 h (p53) or 14.1 +/- 2.2 h (N-ras) for interstrand cross-links. Moreover, peak levels of monoadducts in both genes were observed 2 h after treatment in peripheral leukocytes from patients with multiple myeloma treated with high-dose i.v. melphalan, supported by autologous stem cell transplantation, whereas interstrand cross-links were maximal within 8 h. Of seven patients examined, the three who showed the least levels of DNA damage did not respond to the high-dose melphalan. CONCLUSIONS: This is the first report showing that it is feasible to measure gene-specific DNA damage in a readily accessible tissue of humans exposed to bifunctional alkylating drugs and to examine, at the level of the individual patient, the relationships between the induction/repair of cytotoxic DNA damage and clinical response or long-term complications. PMID- 14555521 TI - Estrogen receptor-beta expression in human testicular germ cell tumors. AB - PURPOSE: Estrogen exposure has been linked to a risk for the development of testicular germ cell cancers. The effects of estrogen are now known to be mediated by estrogen receptor (ER)-alpha and -beta receptor subtypes, but only ER beta has been found in human normal testis. The goal of the present study was to compare the localization and expression levels of these ER subtypes in testicular germ cell cancers (seminomas and nonseminomatous germ cell tumors) with normal testis. For completeness, expression of androgen and progesterone receptors was also investigated. EXPERIMENTAL DESIGN: Immunohistochemistry was used to localize the expression of steroid receptors in 39 archival testicular germ cell cancers and 5 morphologically normal testes. Expression of the steroid receptors at the transcript level was semiquantified by reverse transcription-PCR in 5 paired fresh-frozen specimens of normal and neoplastic testes. RESULTS: ER-alpha was not expressed in the human normal testis. It was also absent in all of the testicular germ cell cancers studied. In contrast, ER-beta was strongly expressed in various germ cells of the normal testis. However, its expression was markedly diminished in seminomas, embryonal cell carcinomas, and in mixed germ cell tumors, at both transcriptional and translational levels. In contrast, ER-beta remained highly expressed in endodermal sinus tumors and teratomas. Progesterone receptor, an estrogen-regulated gene, was localized to spermatagonia of the normal testis, but its expression dramatically reduced in seminomas. With the exception of spermatagonia, androgen receptor was found in all of the germ cells of the normal testis, but, aside from trace staining in 3 of 5 endodermal sinus tumor cells, it was not detected immunohistochemically in any other germ cell cancer. CONCLUSIONS: We confirm expression of ER-beta, but not ER-alpha, in normal testicular cells, suggesting that only the former ER subtype mediates the action of estrogen in the human male gonad. Our results provide the first evidence that only ER-beta is expressed in testicular germ cell tumors. Its expression is down regulated in seminomas and embryonal cell carcinomas but remains high in endodermal sinus tumors and in teratomas. The observed differences in ER-beta expression levels among different testicular germ cell tumors may reflect divergent pathways of differentiation/dedifferentiation of these neoplasms from a common precursor. Collectively, these findings provide a possible mechanistic link between estrogen exposure and testicular cancer risk. PMID- 14555522 TI - FLT3 expressing leukemias are selectively sensitive to inhibitors of the molecular chaperone heat shock protein 90 through destabilization of signal transduction-associated kinases. AB - PURPOSE: We conducted studies to evaluate the hypothesis that FLT3 is a client of heat shock protein (Hsp) 90 and inhibitors of Hsp90 may be useful for therapy of leukemia. EXPERIMENTAL DESIGN: The effects of the Hsp90-inhibitor 17-allylamino 17-demethoxygeldanamycin (17-AAG) on cell growth, expression of signal transduction kinases, apoptosis, FLT3 phosphorylation and interaction with Hsp90 was determined in FLT3(+) human leukemias. RESULTS: We found that FLT3 is included in a multiprotein complex that includes Hsp90 and p23. 17-AAG inhibited FLT3 phosphorylation and interaction with Hsp90. FLT3(+) leukemias were significantly more sensitive to the Hsp90 inhibitors 17-AAG and Herbimycin A in cell growth assays than FLT3-negative leukemias. Cells transfected with FLT3 became sensitive to 17-AAG. Cell cycle inhibition and apoptosis were induced by 17-AAG. Cells with constitutive expression of FLT3, as a result of internal tandem duplication, were the most sensitive; cells with wild-type FLT3 were intermediate in sensitivity, and FLT3-negative cells were the least sensitive. 17 AAG resulted in reduced cellular mass of FLT3, RAF, and AKT. The mass of another Hsp, Hsp70, was increased. The expression level of MLL-AF4 fusion protein was not reduced by 17-AAG in human leukemia cells. CONCLUSIONS: FLT3(+) leukemias are sensitive to 17-AAG and Herbimycin A. 17-AAG inhibits leukemia cells with either FLT3-internal tandem duplication or wild-type FLT3, in part through destabilization of client kinases including FLT3, RAF, and AKT. 17-AAG is potentially useful for therapy of FLT3-expressing leukemias, including the mixed lineage leukemia fusion gene leukemias. PMID- 14555523 TI - Anti-leukemia effect of perillyl alcohol in Bcr/Abl-transformed cells indirectly inhibits signaling through Mek in a Ras- and Raf-independent fashion. AB - PURPOSE: Perillyl alcohol (POH) displays preventive and therapeutic activity against a wide variety of tumor models, and it has been suggested that this might be associated with the ability of POH to interfere with Ras prenylation. POH also selectively induces G(1) arrest and apoptosis in Bcr/Abl-transformed hematopoietic cells. Because signaling through Ras is necessary for Bcr/Abl transformation, we examined whether POH induces its anti-leukemia effect by inhibiting Ras signaling. EXPERIMENTAL DESIGN: The ability of POH to inhibit posttranslational farnesylation and signaling from Ras as well as signaling through the Raf-Mek-Erk cascade was examined in Bcr/Abl-transformed and mock transformed cells and related to the anti-leukemia effect of POH. RESULTS: POH does not affect Ras prenylation or Ras activity, but it blocks signaling downstream of Ras by reversing the state of activation of the Erk kinase, Mek. POH affects Mek activity only when it is added to intact cells. Treatment of either cell lysates or of purified Mek with POH has no effect on Mek activity. Inhibition of the Mek-Erk pathway seems to be related to the POH anti-leukemia effect for the following reasons: (a) the concentration of POH needed to block the Erk pathway, as well the kinetics with which POH inhibits this signaling cascade, both correlate with the anti-leukemia effect of POH; (b) both U0126 (a specific Mek inhibitor) and POH induce similar anti-leukemia effects; and (c) mock-transformed hematopoietic cells are simultaneously resistant to POH anti leukemia effects and inhibition of the Mek-Erk pathway. CONCLUSION: Blocking Mek is sufficient to induce growth arrest and apoptosis in Bcr/Abl-transformed cells; therefore, POH represents a novel small molecule inhibitor of Mek that might be effective for treating Bcr/Abl leukemias. PMID- 14555524 TI - Preclinical evaluation of targeted cytotoxic luteinizing hormone-releasing hormone analogue AN-152 in androgen-sensitive and insensitive prostate cancers. AB - PURPOSE AND EXPERIMENTAL DESIGN: To improve conventional chemotherapy, we developed cytotoxic analogues of luteinizing hormone-releasing hormone (LH-RH), which can be targeted to prostate cancers expressing LH-RH receptors. In view of pending clinical trials on cytotoxic LH-RH analogue AN-152, containing doxorubicin (DOX) linked to [D-Lys(6])-LH-RH, we investigated the effects of AN 152 on tumor growth of s.c. implanted androgen-sensitive LNCaP and MDA-PCa-2b prostate cancers, as well as androgen-independent C4-2 prostate cancers xenografted into the tibiae of nude mice. In the C4-2 study, serum prostate specific antigen (PSA) levels were also measured. LH-RH receptors were analyzed by reverse transcription-PCR and ligand competition assay. We also evaluated whether AN-152 can affect mRNA expression of human epidermal growth factor receptor and HER-2 and -3 oncogenes RESULTS: After 32 days of treatment with AN 152, the growth of LNCaP cancers in castrated nude mice was strongly inhibited by 83% versus intact controls (P < 0.01) and 62% versus castrated controls (P < 0.05). In animals bearing MDA-PCa-2b prostate cancers, therapy with AN-152 for 25 days resulted in a 69% inhibition of tumor growth (P < 0.01 versus controls) and was more effective (P < 0.05) than equimolar doses of DOX or microcapsules of LH RH agonist Decapeptyl. In nude mice bearing intraosseous C4-2 prostate cancers, treatment with AN-152 decreased serum PSA levels (P < 0.01) to 10.3 +/- 3.4 ng/ml from 24.8 +/- 4 ng/ml in controls, whereas DOX had no effect on PSA. The inhibitory effects of AN-152 on C4-2 tumors was accompanied by an increase in apoptosis and a decrease in tumor proliferation. Binding sites for LH-RH and the expression of mRNA for LH-RH receptors were found on s.c. C4-2 and MDA-PCa-2b tumors. The inhibition of MDA-PCa-2b tumors by AN-152 was associated with a significant decrease in mRNA expression for epidermal growth factor receptor, HER 2, and 3. CONCLUSIONS: Our findings suggest that cytotoxic analogue AN-152 could be considered for therapeutic trials in patients with advanced prostate carcinoma. PMID- 14555526 TI - Nonviral cytokine gene therapy on an orthotopic bladder cancer model. AB - PURPOSE: The purpose is to assess cytokine gene transfection in tumor cells and its therapeutic efficacy in an orthotopic mouse bladder cancer model after liposome-mediated gene transfer. EXPERIMENTAL DESIGN: A total of 1 x 10(5) MB49 cells was instilled into the bladder of C57BL/6 mice after electrocautery to establish the tumor model. The plasmids were constructed by inserting the coding sequences for murine IFN-alpha1 and granulocyte macrophage colony-stimulating factor into a plasmid vector pBudCE4.1. Transient transfection was performed using a cationic lipid N-[1-(2,3-dioleoyloxyl)propyl]-N,N,N trimethylammoniummethyl sulfate and methyl-beta-cyclodextrin-solubilized cholesterol. The in vitro expression of cytokines was checked by ELISA. The expression of the transgene in situ was confirmed by immunohistochemistry and 5 bromo-4-chloro-3-indolyl-beta-D-galactopyranoside staining. Mice bearing orthotopic tumors were treated with plasmid DNA/liposome complex by intravesical instillation twice a week for 3 weeks. RESULTS: Superficial bladder tumors were established by intravesical instillation of MB49 into cauterized bladders. The expression level of cytokines in transfected cell lines was increased significantly. In situ gene transfer to bladder tumors was accomplished via intravesical instillation of plasmid DNA/N-[1-(2,3-dioleoyloxyl)propyl]-N,N,N trimethylammoniummethyl sulfate/methyl-beta-cyclodextrin-solubilized cholesterol after a single 2 h in situ transfection. The tumor incidence in the treatment groups was dramatically decreased from 76.9% in the control group to 15.4-30.8% in the treatment groups. CONCLUSIONS: We demonstrated in the orthotopic mouse bladder cancer model that successful inhibition of tumor cell growth could be obtained with cytokine gene therapy. The results suggest that our liposome transfection system appears to be a promising method for gene therapy of bladder cancer in vivo. PMID- 14555525 TI - Stress-related mediators stimulate vascular endothelial growth factor secretion by two ovarian cancer cell lines. AB - PURPOSE: Stress has long been believed to influence carcinogenesis, but little is known about physiological mechanisms that may underlie these effects. We have recently observed lower levels of vascular endothelial growth factor (VEGF) in ovarian cancer patients with greater social support, whereas higher VEGF was found in patients with greater distress. The goal of this study was to examine possible mechanisms underlying these relationships. EXPERIMENTAL DESIGN: The effects of stress-related mediators including norepinephrine (NE), epinephrine, isoproterenol (a nonspecific beta-adrenergic agonist), and cortisol on the production of VEGF by the ovarian cell lines SKOV3 and EG were investigated. RESULTS: NE and isoproterenol significantly enhanced VEGF production by SKOV3 cells, and all three of the adrenergic agonists enhanced VEGF production by EG cells. These effects were blocked by the beta antagonist propranolol, supporting a role for beta-adrenergic receptors in these effects. Reverse transcriptase-PCR studies indicated constitutive expression of beta-1 and beta-2 adrenergic receptors on both cell lines. Effects of cortisol on VEGF production varied according to the specific cell line and dose, with stimulating effects on SKOV3 at pharmacologic doses (1000 nM) and on EG at physiological stress level doses (10 nM), and inhibitory effects on EG at pharmacologic doses. Although priming with cortisol blunted NE-induced VEGF production from both cell lines at 3 h, significant increases in VEGF were still seen. Priming with cortisol enhanced isoproterenol-induced VEGF production from SKOV3. CONCLUSION: These findings provide the first experimental evidence of a pathway by which biobehavioral stress mediators could directly contribute to the progression of ovarian tumors. PMID- 14555527 TI - Metabolic response of the CWR22 prostate tumor xenograft after 20 Gy of radiation studied by 1H spectroscopic imaging. AB - PURPOSE: The ability to determine the spatial and metabolic distribution of prostate cancer is essential in assessing initial stage, prognosis, and treatment efficacy. Current markers of tumor progression such as prostate-specific antigen (PSA) do not provide spatial information about tumor extent or regions of high metabolic activity. EXPERIMENTAL DESIGN: This study used the androgen-dependent CWR22 human prostate tumor xenograft in mice to characterize metabolic, PSA, and tumor volume changes that occurred with untreated growth or radiation therapy (XRT). One cohort of mice was studied as the tumor grew to 400 mm(3), whereas a second cohort was treated with a single 20-Gy fraction of radiation and studied before and 1, 2, and 4 days after XRT. In both cohorts, tumor volume, PSA, and choline:water ratios measured by nuclear magnetic resonance were monitored. RESULTS: The CWR22 tumor had an untreated tumor-doubling time of 2.6 +/- 0.6 days (n = 7). In untreated mice, PSA strongly correlated with tumor volume (P < 0.01, R(2) = 0.99). The untreated tumor cohort had a PSA-doubling time of 3.2 +/- 0.6 days. Administration of 20 Gy produced a regrowth delay of >15.8 +/- 4.8 days (n = 6). PSA values after XRT were not correlated with post-XRT tumor volume (P < 0.20, R(2) = 0.02). A constant level of the choline:water ratio (0.010 +/- 0.001; n = 22, R(2) = 0.007, P < 0.3) was observed during the course of untreated tumor growth. A statistically significant (P < 0.04, one-tailed t test) 42% decrease in the choline:water ratio at 24 h after administration of XRT preceded observable changes in PSA. CONCLUSIONS: Nuclear magnetic resonance spectroscopy provided a method with which to monitor metabolic changes of tumor response to XRT that preceded and predicted PSA and tumor volume changes. PMID- 14555528 TI - Drug interactions between the proteasome inhibitor bortezomib and cytotoxic chemotherapy, tumor necrosis factor (TNF) alpha, and TNF-related apoptosis inducing ligand in prostate cancer. AB - PURPOSE: Proteasome inhibition has been shown to be an effective anticancer therapy in many tumor models, including prostate cancer. We sought to identify drug interactions between the proteasome inhibitor bortezomib and other apoptotic stimuli, including cytotoxic chemotherapy and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). In addition, we wanted to gain insight into the role of nuclear factor kappaB inhibition as a mediator of bortezomib cytotoxic effects. EXPERIMENTAL DESIGN: Prostate cancer cell lines (LNCaP, LAPC4, CL1, and DU145) were treated with bortezomib and apoptotic stimuli (TRAIL, chemotherapy, and tumor necrosis factor alpha), alone or in combination. Apoptosis and cell viability were measured, and median effect/combination index analyses were used to quantitate drug interactions. Nuclear factor kappaB activity at baseline and in response to drug treatment was determined by gel shift and reporter gene assays. RESULTS: Bortezomib induced cell death of androgen-dependent (LNCaP and LAPC4) and androgen-independent (CL1 and DU145) prostate cancer cell lines, although androgen-dependent cells were more sensitive to proteasome inhibition. Bortezomib synergized with TRAIL and tumor necrosis factor alpha to induce death in both androgen-dependent and androgen-independent cells. CONCLUSIONS: Bortezomib and TRAIL represent a synergistic drug combination that warrants further evaluation in in vivo models of prostate cancer. PMID- 14555529 TI - Changes in tumor vascularization after irradiation, anthracyclin, or antiangiogenic treatment in nitrosomethyl ureas-induced rat mammary tumors. AB - PURPOSE: Changes in tumor vascularization may be involved in tumor regression after anticancer treatments. We therefore studied the relationship between tumor vascularization and tumor response according to treatment by irradiation (RT), epirubicin (EPI), or antiangiogenic agent TNP-470 in a nitrosomethyl-ureas induced rat mammary tumor model by measuring the changes in tumor blood flow using high-frequency Power-Doppler sonography. EXPERIMENTAL DESIGN: Mammary tumors were induced in female Sprague-Dawley rats by a single s.c. injection of nitrosomethyl-ureas. After tumor areas reached 2 cm(2), the animals received four weekly injections of epirubicin (EPI group), or a single fraction of 18 Gy (RT group), or six injections of TNP-470 within 12 days (TNP group), or both (RT combined with TNP-470, RT+TNP group). Power-Doppler sonography quantification of tumor vascularization (PDI) was performed before and 12 days after initiation of treatment. Tumor shrinkage was later evaluated and compared with the early changes in PDI values. RESULTS: Compared with the control group, EPI induced an arrest in tumor growth. A similar effect was obtained with TNP-470. There was a decrease in tumor area after RT, but administration of TNP-470 combined with RT did not further enhance this effect. Changes in tumor area paralleled changes in PDI in the EPI group. Furthermore, changes in PDI 7 days after RT were associated with further tumor change in the RT groups, whereas they were independent of the antitumor effect of TNP-470. CONCLUSIONS: Changes in functional tumor vascularization evolution appeared to be closely associated with tumor regression after anticancer treatment. PMID- 14555530 TI - Adenovirus-mediated ribonucleotide reductase R1 gene therapy of human colon adenocarcinoma. AB - Ribonucleotide reductase is the enzyme responsible for the reduction of ribonucleotides to their corresponding deoxyribonucleotides for DNA synthesis. Ribonucleotide reductase is a multisubunit complex containing two polypeptides, R1 and R2. In addition to catalytic and allosteric regulatory functions, the R1 subunit appears to act as a novel tumor suppressor. Previous studies demonstrated that overexpression of mouse R1 resulted in suppression of tumorigenicity and metastatic potential, whereas expression of antisense RNA, complementary to R1 mRNA, increased anchorage-independent growth of ras-transformed NIH 3T3 cells. The current study investigated the potential of R1 gene therapy for human cancer using a recombinant adenovirus encoding the human R1 gene (rAd5-R1). Recombinant viruses were constructed by FLP-mediated site-specific recombination and demonstrated high infectivity of a human colon carcinoma cell line (Colo320 HRS), as assessed by expression of a viral encoded beta-Gal gene (rAd5-LacZ). R1mRNA and protein were overexpressed in Colo320 HRS cells infected with rAd5-R1 compared with untreated or rAd5-LacZ-infected cells. Infection with rAd5-R1 inhibited Colo320 HRS cell proliferation, in vitro, in a time- and dose-dependent manner. When Colo320 HRS cells were treated with rAd5-R1, before injection into CD-1 mice, there was complete inhibition of tumor growth compared with treatment with rAd5-LacZ. Furthermore, intratumoral injection of rAd5-R1 into Colo320 HRS tumor xenografts inhibited tumor growth in CD-1 mice compared with rAd5-LacZ treated mice (P = 0.0001). These results demonstrate gene-specific antitumor effects of R1 and suggest that rAd5-R1 gene therapy has the potential to improve currently available treatments for colon cancer. PMID- 14555531 TI - Is interferon gamma one key of metastatic potential increase in human bladder carcinoma? AB - PURPOSE: IFN-gamma is detected in the urine of bladder cancer patients after intravesical bacillus Calmette Guerin instillation. Because it acts in the anticancer process, we studied its cellular and molecular mechanisms of action on human bladder cancer cell lines. RESULTS: IFN-gamma (>5 ng.ml(-1))(>400 IU.ml( 1)) inhibited the growth of bladder cancer cell lines and modified the expression of the tumor-associated markers tissue-type plasminogen activators, Plasminogen activator inhibitor-2, urokinase plasminogen activator receptor, colony stimulating factor 1, intercellular adhesion molecule 1, and class II MHC. Interestingly, IFN-gamma-induced apoptosis of the low-grade bladder cancer cell lines (RT4/G1 and RT112/G2) related to a cleavage of caspases 1, 8, and 9. This process was inhibited by the phosphatidylinositol 3'-kinase inhibitor (LY294002) and the protein synthesis inhibitor (cycloheximide). Moreover, low doses of IFN gamma (<5 ng.ml(-1))(<400 IU.ml(-1)) increased the resistance to the cytotoxic effect of tumor necrosis factor alpha in the RT112 cells but not in the RT4 cells. This acquired resistance was associated with morphological changes and with an increase of the cell migration and scattering. CONCLUSIONS: We demonstrated that in the low-grade bladder cancer cell lines, the effect of IFN gamma was dose dependent: high doses (>5 ng.ml(-1)) induced apoptosis of RT4 and RT112 cells, whereas low doses (<5 ng.ml(-1)) induced a resistance to the cytotoxic effect of tumor necrosis factor alpha and increase the metastatic potential of the RT112 cells. Therefore, we propose that a similar phenomenon could participate to the immunotherapy failure observed during tumor progression of bladder cancer. PMID- 14555532 TI - Effects of the proteasome inhibitor, bortezomib, on apoptosis in isolated lymphocytes obtained from patients with chronic lymphocytic leukemia. AB - PURPOSE: Bortezomib is a peptide boronic acid inhibitor of the proteasome developed for cancer therapy. The compound is being evaluated currently in Phase II and III clinical trials. Here we characterized the effects and mechanisms of action of bortezomib in cells obtained from patients with chronic lymphocytic leukemia (CLL). EXPERIMENTAL DESIGN: We exposed isolated CLL lymphocytes from >100 patients to various concentrations of bortezomib or other proapoptotic stimuli, and measured DNA fragmentation by propidium iodide staining and flow cytometry. We characterized the effects of bortezomib on release of apoptosis associated mitochondrial factors and measured downstream effects on caspase activation using a fluorogenic substrate cleavage assay. We assessed potential effects of the drug on inhibitor of apoptosis protein family apoptosis inhibitors by immunoblotting. Finally, we quantified the effects of bortezomib on apoptosis in 5 patients on a Phase II clinical trial. RESULTS: Bortezomib stimulated apoptosis more rapidly than positive controls (glucocorticoid and fludarabine), although substantial heterogeneity was noted with respect to the concentration of drug required to induce cell death. Bortezomib-induced apoptosis was associated with release of SMAC, apoptosis-inducing factor, and cytochrome c from mitochondria, but the drug did not affect levels of inhibitor of apoptosis protein family cell death inhibitors. Levels of apoptosis were marginally elevated in CLL cells obtained from 2 of 5 fludarabine-refractory patients treated with bortezomib in vivo. CONCLUSION: Our data confirm that bortezomib, like other proteasome inhibitors, has proapoptotic activity in CLL cells. PMID- 14555533 TI - Inhibition of intracerebral glioblastoma growth by local treatment with the scatter factor/hepatocyte growth factor-antagonist NK4. AB - PURPOSE: Scatter factor/hepatocyte growth factor (SF/HGF) and its tyrosine kinase receptor MET are strongly up-regulated in malignant gliomas. The SF/HGF-MET system contributes to glioma invasion and angiogenesis via autocrine and paracrine mechanisms. We analyzed whether local treatment with NK4, an antagonistic fragment of SF/HGF, could inhibit glioma growth in vivo. EXPERIMENTAL DESIGN: A guide-screw system was used to implant tumor cells intracerebrally and to perform therapeutic injections. Mice received daily intratumoral injections of NK4 or buffer as of day 1 or 7 after tumor cell injection until day 20. Functional effects of NK4 on glioma and endothelial cells were analyzed in vitro. RESULTS: Tumor volume was reduced by 61.1% in mice treated with NK4 compared with controls when treatment was initiated on day 1 (P < 0.05) and by 61.4% when treatment was initiated on day 7 (P < 0.001). Intratumoral microvessel density was reduced by 64.9% when treatment started on day 1 and by 36.7% when it started on day 7. The proliferative activity of the tumor cells was reduced by >30% regardless of when NK4-treatment was initiated. The apoptotic fraction of tumor cells was increased 2-fold and 1.5-fold when animals were treated with NK4 as of day 1 or day 7, respectively. In vitro, NK4 inhibited SF/HGF-induced glioblastoma, and endothelial cell migration and proliferation in a dose-dependent fashion. CONCLUSION: NK4 inhibits glioblastoma growth in vivo, most likely via antimitogenic, antimotogenic, proapoptotic, and antiangiogenic mechanisms. Given the strong up-regulation of SF/HGF and MET in human malignant gliomas, NK4 holds promise as a direct interstitial therapeutic agent for these fatal tumors. PMID- 14555534 TI - Flavopiridol potently induces small cell lung cancer apoptosis during S phase in a manner that involves early mitochondrial dysfunction. AB - PURPOSE: Accumulating evidence indicates that small cell lung cancer (SCLC) is defective in many of the regulatory mechanisms that control cell cycle progression. The purpose of this study was to determine the effects of flavopiridol, a pan-cyclin-dependent kinase inhibitor, on growth and apoptosis of SCLC cell lines. EXPERIMENTAL DESIGN: Cell growth was monitored using 3 (4,5dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide (MTT) and clonogenic assays. Induction of apoptosis was assessed using multiple assays, including flow cytometric determination of DNA content and mitochondrial membrane potential, terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL), and Western blot analysis of procaspase 3 and poly(ADP-ribose) polymerase cleavage. RESULTS: Flavopiridol induced growth inhibition and cytotoxicity in multiple SCLC cell lines, with an IC(50) of 50-100 nM and an LD(50) of 150-200 nM in 72-h MTT assays. The cytotoxicity seen in the MTT assay proved to be apoptosis by several criteria. Interestingly, inhibition of caspase activation with the caspase inhibitor Boc-Asp(OMe)-CH(2)F reduced TUNEL labeling by 40% but did not have any effect on the loss of mitochondrial membrane potential (detected as early as 4 h after drug exposure) or cytotoxicity in MTT assays. These results suggest that the primary event in flavopiridol-induced apoptosis involves induction of mitochondrial dysfunction. Cells synchronized with aphidicolin at the G(1)-S border and treated with flavopiridol during S phase showed a marked increase in apoptosis compared with an asynchronous population or a population treated during G(2)-M. Despite the increased apoptosis, a significant proportion of synchronized cells proceeded through S, G(2)-M, and into G(1) phase in the presence of flavopiridol, demonstrating that a high-grade cell cycle arrest is not required for apoptosis. Cells synchronized at the G(1)-S border treated with a short exposure to flavopiridol also showed more than a 10-fold decrease in clonogenicity compared with asynchronous cells treated identically. CONCLUSIONS: Taken together, these data demonstrate that flavopiridol potently and selectively induces SCLC apoptosis preferentially during S phase, in a manner that involves early mitochondrial dysfunction without a requirement for a high-grade block to cell cycle progression. Furthermore, clonogenicity data suggests that prior S phase synchronization could be a highly effective way of enhancing the efficacy of bolus or short infusions of flavopiridol in the clinical setting. PMID- 14555535 TI - Targeted liposomal c-myc antisense oligodeoxynucleotides induce apoptosis and inhibit tumor growth and metastases in human melanoma models. AB - PURPOSE: Melanoma is a highly malignant and increasingly common tumor. Because the cure rate of metastatic melanoma by conventional treatment is very low, new therapeutic approaches are needed. We previously reported that coated cationic liposomes (CCL) targeted with a monoclonal antibody against the disialoganglioside (GD(2)) and containing c-myb antisense oligodeoxynucleotides (asODNs) resulted in a selective inhibition of the proliferation of GD(2) positive neuroblastoma cells in vitro. EXPERIMENTAL DESIGN: Here, we tested the in vivo antitumor effects of this novel antisense liposomal formulation by targeting the c-myc oncogene on melanoma, a neuroectodermal tumor sharing with neuroblastoma the expression of GD(2). RESULTS: Our methods produced GD(2) targeted liposomes that stably entrapped 90% of added c-myc asODNs. These liposomes showed a selective binding for GD(2)-positive melanoma cells in vitro. Melanoma cell proliferation was inhibited to a greater extent by GD(2)-targeted liposomes containing c-myc asODNs (aGD(2)-CCL-myc-as) than by nontargeted liposomes or free asODNs. The pharmacokinetic results obtained after i.v. injection of [(3)H]-myc-asODNs, free or encapsulated in nontargeted CCLs or GD(2) targeted CCLs, showed that free c-myc-asODNs were rapidly cleared, with less than 10% of the injected dose remaining in blood at 30 min after injection. c-myc asODNs encapsulated within either CCL or aGD(2)-CCL demonstrated a more favorable profile in blood, with about 20% of the injected dose of each preparation remaining in vivo at 24 h after injection. In an in vivo melanoma experimental metastatic model, aGD(2)-CCL-myc-as, at a total dose of only 10 mg of asODN per kilogram, significantly inhibited the development of microscopic metastases in the lung compared with animals treated with myc-asODNs, free or entrapped in nontargeted liposomes, or aGD(2)-CCL encapsulating scrambled asODNs (P < 0.01). Moreover, mice bearing established s.c. human melanoma xenografts treated with aGD(2)-CCL-myc-as exhibited significantly reduced tumor growth and increased survival (P < 0.01 versus control mice). The mechanism for the antitumor effects appears to be down-regulation of the expression of the c-myc protein and interruption of c-myc-mediated signaling: induction of p53 and inhibition of Bcl 2 proteins, leading to extensive tumor cell apoptosis. CONCLUSION: These results suggest that inhibition of c-myc proto-oncogene by GD(2)-targeted antisense therapy could provide an effective approach for the treatment of melanoma in an adjuvant setting. PMID- 14555536 TI - Retinoid receptor-dependent and independent biological activities of novel fenretinide analogues and metabolites. AB - Fenretinide (4-HPR) is a retinoid analogue with antitumor and chemopreventive activities. In addition to 4-HPR, there are several other new phenylretinamides bearing hydroxyl, carboxyl, or methoxyl residues on carbons 2, 3, and 4 of the terminal phenylamine ring [N-(2-hydroxyphenyl)retinamide (2-HPR), N-(3 hydroxyphenyl)retinamide, N-(2-carboxyphenyl)retinamide, N-(3 carboxyphenyl)retinamide, N-(4-carboxyphenyl)retinamide, and N-(4 methoxyphenyl)retinamide (4-MPR) ]. It is hypothesized that these agents can act independent of the nuclear retinoid receptor pathway. To test this hypothesis directly, we have analyzed the activity of these phenylretinamides in vitro on a panel of F9 murine embryonal carcinoma cell lines, which includes wild-type (F9 WT) and mutant cells that have disrupted genes for both retinoid X receptor alpha and retinoic acid receptor gamma retinoid receptors (F9-KO). The F9-KO cells lack almost all measurable response to all-trans-retinoic acid, the primary biologically active retinoid. Two distinct effects of retinamides were identified. The first is a rapid, dose-dependent induction of cell growth inhibition (reduced cell viability), and the second is a slower induction of differentiation and accumulation of cells in the G(1) phase of the cell cycle that was observed with a concentration of 1 micro M, for only those phenylretinamides bearing charged (hydroxyl or carboxyl) groups on the terminal phenylamine ring. The induction of differentiation and G(1) accumulation was only observed in the F9-WT cells, indicating that this effect is receptor-dependent. 4 MPR, a major metabolite of 4-HPR, lacks a charged group on the terminal phenylamine ring and did not induce retinoid receptor-dependent effects, but did induce cell growth inhibition. Thus, 4-MPR may play a role in the clinical activity of 4-HPR. This study further reveals the mechanism of action of these novel phenylretinamides and supports continued investigation into their development as chemopreventive drugs. PMID- 14555537 TI - Selective estrogen receptor modulators as inhibitors of repopulation of human breast cancer cell lines after chemotherapy. AB - PURPOSE: Repopulation of surviving tumor cells between courses of chemotherapy might lead to effective drug resistance. Here we study inhibition of repopulation of hormone-responsive human breast cancer cell lines by selective estrogen receptor (ER) modulators (SERMs) during courses of chemotherapy. EXPERIMENTAL DESIGN: Hormone responsive breast cancer cell lines MCF-7 and T47D, and the ER- cell line MDA-231, were treated with either 4-hydroxy tamoxifen (4OHT) or arzoxifene during weekly courses of treatment with 5-fluorouracil (5-FU) or methotrexate (MTX). Clonogenic assays were performed to determine the overall survival of tumor cells after treatment with the SERMs alone, after one to three doses of 5-FU or MTX alone, and after 5-FU or MTX followed by each of the SERMs. RESULTS: Both SERMs inhibited the growth of ER+ cells MCF-7 and T47D but had no effect on the ER-cell line MDA-231. Arzoxifene was more effective than 4OHT. Between courses of treatment with either 5-FU or MTX, repopulation of ER+ cells was specifically inhibited by the SERMs, whereas repopulation of ER- MDA-231 was not affected. CONCLUSIONS: Arzoxifene and 4OHT can inhibit specifically the repopulation of ER+ breast cancer cells between courses of chemotherapy. Scheduling of short-acting SERMs between courses of chemotherapy has the potential to improve therapeutic index. PMID- 14555538 TI - Synergistic activity of recombinant human endostatin in combination with adriamycin: analysis of in vitro activity on endothelial cells and in vivo tumor progression in an orthotopic murine mammary carcinoma model. AB - PURPOSE: Current combination treatment strategies in malignancy are designed to evaluate the use of cytotoxic drugs and antiangiogenic agents. Endostatin, a fragment of collagen XVIII, specifically inhibits proliferation, migration, and differentiation of endothelial cells in vitro as well as angiogenesis and tumor progression in in vivo models. In this study, we determine the antitumor effect of rhEndostatin administered alone or in combination with Adriamycin against established orthotopic murine mammary carcinoma. EXPERIMENTAL DESIGN: Mice bearing orthotopically established DA-3 mammary adenocarcinoma tumors received varying doses of rhEndostatin alone and in combination with Adriamycin to assess tumor growth inhibition. Additional studies of this in vivo combination included a determination of Adriamycin-induced cardiotoxicity and in vitro effects on human umbilical vein endothelial cell proliferation and cord formation. RESULTS: For single-agent activity, optimal tumor growth inhibition was observed after s.c. administration of 50 mg/kg/day rhEndostatin or 5 mg/kg Adriamycin injected i.v. every 4 days. Combination of Adriamycin with optimal or suboptimal doses of rhEndostatin resulted in synergistic inhibition of DA-3 tumor growth. Importantly, unlike other antiangiogenic agents, rhEndostatin did not exacerbate the cardiotoxicity of Adriamycin. The synergistic interaction between rhEndostatin and Adriamycin was also observed in vitro for inhibition of human umbilical vein endothelial cell proliferation and inhibition of cord formation. CONCLUSIONS: These data suggest that the synergy observed with rhEndostatin in combination with Adriamycin is exerted at the level of the endothelial cell and can result in enhanced tumor growth inhibition. The potential benefit of Adriamycin used in combination with rhEndostatin is being considered for clinical evaluation. PMID- 14555539 TI - Control of COX-2 gene expression through peroxisome proliferator-activated receptor gamma in human cervical cancer cells. AB - PURPOSE: The peroxisome proliferator-activated receptor-gamma (PPARgamma), a ligand-dependent transcription factor belonging to the family of nuclear receptors, has been implicated in the control of cyclooxygenase (COX) 2 expression in some tissue, although the exact mechanism(s) of this activity has not been elucidated. In this study we explored the possible mechanism(s) of control of COX-2 gene expression through PPARgamma signaling in human cervical cancer. EXPERIMENTAL DESIGN: Using primary human cervical tissues and the CaSki human cervical cancer cell line, we assayed for PPARgamma and COX-2 mRNA expression by reverse transcription-PCR. Nuclear protein binding activities to three response elements located in the COX-2 promoter [nuclear factor kappaB (NFkappaB), cyclic AMP response element, and activator protein (AP)-2] were measured by gel mobility shift assays. We used transient transfection assays with COX-2 promoter reporter gene constructs to determine the regulatory sites in this promoter, which mediates PPARgamma regulation of COX-2 activity. RESULTS: We showed, for the first time, that primary human cervical cancer tissues express PPARgamma. Using CaSki cells, we demonstrated that COX-2 and PPARgamma mRNA levels were inversely regulated by PPARgamma ligands in that these compounds up regulated PPARgamma but down-regulated COX-2. In contrast, epidermal growth factor (EGF), a potent activator of COX-2, decreased PPARgamma mRNA levels. This down-regulation of PPARgamma mRNA by EGF was blocked in the presence of NS-398, a selective COX-2 inhibitor. PPARgamma ligands suppressed the binding activities of AP-1 (binding to CRE) and NFkappaB but not AP-2. Transient transfection results indicated that EGF stimulated whereas PPARgamma ligands inhibited COX-2 promoter (-327/+59) activity. This effect by PPARgamma ligands on the COX-2 promoter was blocked when the CRE, but not the NFkappaB, binding site was mutagenized. CONCLUSION: Cervical cancer cells express readily detectable levels of PPARgamma. There is reciprocal negative regulation between COX-2 and PPARgamma signaling in human cervical cancer cells. The ability of PPARgamma ligands to inhibit COX-2 appears to be mediated predominantly through inhibition of AP-1 protein binding to the CRE site in the COX-2 promoter. PMID- 14555540 TI - Treatment of ifosfamide encephalopathy with intravenous thiamin. PMID- 14555541 TI - The spectrum of smoking-related interstitial lung disorders: the never-ending story of smoke and disease. PMID- 14555542 TI - Idiopathic pulmonary fibrosis: the inflammation hypothesis revisited. PMID- 14555543 TI - Primary graft failure: who is at risk? PMID- 14555544 TI - Interaction of angiotensin-converting enzyme inhibition and aspirin in congestive heart failure: long controversy finally resolved? PMID- 14555545 TI - Hetastarch in perioperative volume expansion. PMID- 14555546 TI - Dosing inhaled steroids in asthma: is once-a-day administration effective? PMID- 14555547 TI - The overlap between respiratory bronchiolitis and desquamative interstitial pneumonia in pulmonary Langerhans cell histiocytosis: high-resolution CT, histologic, and functional correlations. AB - OBJECTIVES: To characterize the histologic, high-resolution CT (HRCT), and functional correlates of respiratory bronchiolitis (RB)/desquamative interstitial pneumonia (DIP)-like changes in biopsy specimens of adults with pulmonary Langerhans cell histiocytosis (PLCH). METHODS: We retrospectively identified 14 adult patients with histologically proven PLCH (all smokers) in whom both biopsy specimens and HRCT were available for review. The presence and extent of RB/DIP like changes on lung biopsy specimens of patients with PLCH were correlated with total pack-years of smoking, lung function variables, and the presence of ground glass attenuation on HRCT. RESULTS: Varying degrees of RB/DIP-like changes were identified in all biopsy specimens. The extent of involvement with RB/DIP-like changes correlated with the cumulative exposure to cigarettes smoked at the time of biopsy (r = 0.61, p = 0.03). Ground-glass attenuation was detected in three HRCTs, two of which had extensive RB/DIP-like changes (> or = 70% involvement) on histology, suggesting that substantial smoking-induced RB/DIP-like changes may be the cause of ground-glass attenuation on the HRCT of patients with PLCH. No correlation was found between the extent of RB/DIP-like changes and total lung capacity, FEV(1), or diffusion capacity of the lung for carbon monoxide. CONCLUSIONS: RB/DIP-like changes are exceedingly common in PLCH, may be sufficiently severe to cause the appearance of ground-glass attenuation on HRCT, and correlate with the cumulative exposure to cigarettes smoked. This study provides additional evidence that PLCH, RB, and DIP form a spectrum of interstitial patterns of lung injury to cigarette smoke. PMID- 14555548 TI - The increase in serum soluble ST2 protein upon acute exacerbation of idiopathic pulmonary fibrosis. AB - BACKGROUND: The human ST2 gene can be specifically induced by growth stimulation in fibroblastic cells, and the soluble ST2 protein (ST2) is expressed preferentially in T-helper type 2 (Th2) cells. Furthermore, ST2 is induced by proinflammatory stimuli such as tumor necrosis factor-alpha and interleukin-1 beta. It has been reported that the inflammatory response in idiopathic pulmonary fibrosis (IPF) is thought to be associated with proinflammatory cytokines and Th2 immune response. STUDY OBJECTIVE: The objective of this study was to evaluate the relevance of the serum ST2 levels in the pathogenesis of IPF. DESIGN: Retrospective study. SETTING: Inpatients in a college hospital. PARTICIPANTS: Forty-nine patients with IPF admitted to our hospital 64 times: 36 patients were admitted once, 11 patients were admitted twice, and 2 patients were admitted three times. The participants also included 200 healthy control volunteers. MEASUREMENTS AND RESULTS: Among 64 events in 49 patients with IPF, 50 of the events occurred in a stable state, and 14 events occurred during acute exacerbation. An acute exacerbation of IPF was defined as an accelerated phase of IPF. The serum ST2 levels were measured by enzyme-linked immunosorbent assay. The serum levels of ST2 in the stable state group did not differ from those in the healthy control group, while the serum levels of ST2 in the acute exacerbation group were significantly higher than those in the stable state group or the healthy control group (p < 0.001, acute exacerbation group vs stable state group or healthy control group; acute exacerbation group, 2.76 +/- 0.56 ng/mL; stable state group, 0.44 +/- 0.07 ng/mL; healthy control group, 0.42 +/- 0.03 ng/mL). Furthermore, serum ST2 statistically correlated with lactate dehydrogenase (r = 0.344, p = 0.005) and C-reactive protein (r = 0.496, p < 0.001), and inversely correlated with PaO(2) (r = - 0.356, p = 0.018) and the percentage of predicted vital capacity (r = - 0.346, p = 0.026). CONCLUSIONS: These results suggest that ST2 protein may increase in the serum, reflecting severity in the inflammatory process and Th2 immune response in the IPF lung. PMID- 14555549 TI - Radiologic findings are strongly associated with a pathologic diagnosis of usual interstitial pneumonia. AB - PURPOSE: To determine which clinical and radiologic findings are independently associated with a pathologic diagnosis of usual interstitial pneumonia (UIP). METHODS: We recently reported, using a prospective, multicenter study of patients suspected of having idiopathic interstitial pneumonia (IIP), that a confident diagnosis of UIP made by experienced radiologists was correct in 95% of cases. In the current article, we further analyzed data from this study. Ninety-one patients were entered into the study. Clinical, physiologic, chest radiographic, and CT features were prospectively recorded, and analyzed using univariate and multivariate logistic regression analysis to compare the patients with a histologic diagnosis of UIP with those who received other pathologic diagnoses. RESULTS: Fifty-four of 91 patients (59%) received a pathologic diagnosis of UIP. The following features recorded at the referring clinical centers were associated with a pathologic diagnosis of UIP on multivariate analysis: lower-lobe honeycombing on high-resolution CT (HRCT) [odds ratio, 11.45], radiographic findings consistent with UIP (odds ratio, 5.73), elevated ratio of FEV(1) to FVC (odds ratio, 4.8), and absence of smoking history (odds ratio, 0.19). On multivariate analysis of specific HRCT features recorded by four experienced chest radiologists, lower-lung honeycombing (odds ratio, 5.36) and upper-lung irregular lines (odds ratio, 6.28) were the only independent predictors of UIP. Using only these two factors, a diagnosis of UIP could be established with a sensitivity of 74%, a specificity of 81%, and a positive predictive value of 85%. CONCLUSION: In patients presenting with a clinical syndrome suggestive of IIP, CT findings of lower-lung honeycombing and upper-lung irregular lines are most closely associated with a pathologic diagnosis of UIP. PMID- 14555550 TI - A longitudinal study of lung function in nonsmoking patients with rheumatoid arthritis. AB - STUDY OBJECTIVE: s: Patients with rheumatoid arthritis (RA) have a high prevalence of pulmonary function test (PFT) abnormality, but the long-term significance of this is unknown. We performed a longitudinal study of pulmonary function in asymptomatic, nonsmoking patients with active RA requiring disease modifying drugs. We looked for temporal change in lung function and characteristics that would predict subsequent development of PFT abnormality or respiratory symptoms. METHODS: In 1990, 52 patients (44 women; age range, 29 to 78 years; median, 56 years) underwent clinical assessment (drug history, RA severity, immunologic, and inflammatory markers) and PFTs (spirometry, body plethysmography, gas transfer). PFT results were expressed as standardized residuals (SRs). Thirty-eight patients were reassessed in 2000. A self administered questionnaire was used to identify respiratory symptoms. RESULTS: The prevalence of pulmonary function abnormality was higher than expected compared with a reference population, but there was no significant increase in number over 10 years (8.7% in 1990 and 8.8% in 2000). When assessed by group means and compared with reference values, reduced diffusing capacity of the lung for carbon monoxide (DLCO) and increased ratio of residual volume (RV) to total lung capacity (TLC) [RV/TLC] were the only abnormalities to develop over the study period (mean DLCO in 2000, - 0.47 SR; 95% confidence interval [CI], - 0.91 to - 0.01; RV/TLC, 0.49 SR; 95% CI, 0.13 to 0.84). However, rates of change of pulmonary function variables were not significantly different from zero. Logistic regression did not identify any meaningful relationship between disease characteristics and PFT abnormality. CONCLUSIONS: Asymptomatic patients with RA have a higher prevalence of PFT abnormality than expected, but these do not increase in number over time. We did not identify any patient or disease-specific characteristic that could predict the development of respiratory disease in patients with RA. Analysis using percentage of predicted values, rather than SRs, is misleading as it exaggerates the extent of abnormality present. Abnormal lung function is a common and probably benign finding in nonsmoking, asymptomatic patients with RA. PMID- 14555551 TI - Clinical risk factors for primary graft failure following lung transplantation. AB - STUDY OBJECTIVE: s: Primary graft failure (PGF) is a devastating acute lung injury syndrome following lung transplantation. We sought to identify donor, recipient, and operative risk factors for its development. DESIGN: We conducted a cohort study of 255 consecutive lung transplant procedures performed between October 1991 and July 2000. We defined PGF as follows: (1) diffuse alveolar opacities exclusively involving allograft(s) and developing within 72 h of transplant, (2) a ratio of PaO(2) to fraction of inspired oxygen < 200 beyond 48 h postoperatively, and (3) no other secondary cause of graft dysfunction identified. Risk factors were assessed individually and adjusted for confounding using multivariable logistic regression models. SETTING: Tertiary-care academic medical center. RESULTS: The overall incidence was 11.8% (95% confidence interval [CI], 7.9 to 15.9). Following multivariable analysis, the risk factors independently associated with development of PGF were as follows: a recipient diagnosis of primary pulmonary hypertension (PPH; adjusted odds ratio [OR], 4.52; 95% CI, 1.29 to 15.9; p = 0.018), donor female gender (adjusted OR, 4.11; 95% CI, 1.17 to 14.4; p = 0.027), donor African-American race (adjusted OR, 5.56; 95% CI, 1.57 to 19.8; p = 0.008), and donor age < 21 years (adjusted OR, 4.06; 95% CI, 1.34 to 12.3; p = 0.013) and > 45 years (adjusted OR, 6.79; 95% CI, 1.61 to 28.5; p = 0.009). CONCLUSIONS: Recipient diagnosis of PPH, donor African-American race, donor female gender, and donor age are independently and strongly associated with development of PGF. PMID- 14555552 TI - Posttransplant lymphoproliferative disorder: incidence, presentation, and response to treatment in lung transplant recipients. AB - INTRODUCTION: Posttransplant lymphoproliferative disorder (PTLD) is a relatively infrequent but devastating complication that occurs after solid-organ transplantation. Although the optimal treatment for this condition is unknown, rituximab, a murine/human chimeric monoclonal antibody, has shown promise in the treatment of PTLD. In this report, we define the incidence, clinical features at presentation, and response to treatment of all cases of PTLD observed at our institution over a 10-year period, including four patients who received treatment with rituximab. METHODS: A review of all patients who underwent lung or heart lung transplant at Duke University from 1992 to 2002 was performed (n = 400), and demographic and clinical outcome data were extracted. RESULTS: PTLD was observed in 10 of 400 patients (2.5%). Patients who acquired PTLD were predominantly > 55 years old (8 of 10 patients) and with a native disease of COPD (7 of 10 patients). Diagnosis of PTLD was made a median of 343 days after transplant. The type of transplant and Epstein-Barr virus (EBV) status prior to transplant did not appear to influence the risk for PTLD. Patients presented with thoracic organ involvement (7 of 10 patients), extrapulmonary disease (2 of 10 patients), or both (1 of 10 patients). Histologic subtypes included polymorphic B cell (n = 4), monomorphic B cell (n = 3), B cell without further classification (n = 2), and anaplastic T cell (n = 1). Only one patient responded to reduced immunosuppression alone. Patients treated with surgery or radiation (n = 2) or rituximab (n = 4) had favorable responses to therapy. Both patients treated with chemotherapy died related to complications of treatment and PTLD. CONCLUSIONS: Presentation and histologic appearance of PTLD varies considerably among lung transplant recipients. PTLD was more frequent among older patients with COPD, regardless of pretransplant EBV serology. Rituximab appears effective as a first line therapy for PTLD, but additional studies are needed in order to define its efficacy and side effect profile in this population of patients. PMID- 14555553 TI - Aspirin does not adversely affect survival in patients with stable congestive heart failure treated with Angiotensin-converting enzyme inhibitors. AB - BACKGROUND: Experimental studies and retrospective analyses of mortality trials with angiotensin-converting enzyme inhibitors (ACE-Is) have suggested that aspirin may reduce the beneficial effect of these drugs. The aim of this study was to assess a possible detrimental effect of aspirin on survival in stable patients with left ventricular systolic dysfunction who had congestive heart failure and had been treated with ACE-Is. METHODS AND RESULTS: We performed a retrospective analysis in 755 consecutive stable patients with left ventricular systolic dysfunction. A Cox regression model was used to select independent predictors of survival and to test for a possible interaction between aspirin and ACE-Is with an adjustment to differences in clinical characteristics in subgroups of patients. Of the 755 patients, 328 (43.4%) had proven ischemic cardiomyopathy, 693 patients (91.8%) were receiving ACE-Is, and 317 patients were receiving aspirin (mean [+/- SD] dose, 183 +/- 65 mg/d; 74% of the patients receiving < or = 200 mg/d). During a median follow-up period of 1,996 days, there were 273 cardiac-related deaths, 14 urgent transplantations, 71 nonurgent transplantations, and 46 noncardiac-related deaths, and 3 patients were lost to follow-up. The cardiovascular mortality rates were 11.5% and 19.0%, respectively, at 1 and 2 years. There were no interactions among aspirin, ACE-Is, and survival in the overall population (p = 0.21), or in subgroups of patients with ischemic cardiomyopathy (p = 0.41) or with nonischemic cardiomyopathy (p = 0.74). CONCLUSIONS: In this population of stable patients with left ventricular systolic dysfunction, our retrospective analysis did not demonstrate any interaction between the use of aspirin and survival in patients receiving ACE-Is. PMID- 14555554 TI - Coronary sinus catheter placement: assessment of placement criteria and cardiac complications. AB - STUDY OBJECTIVES: To evaluate the placement and complications of a coronary sinus (CS) catheter in human subjects. DESIGN: Sixty-two CS catheters inserted in patients scheduled for coronary artery bypass graft surgery (CABG). SETTING: University hospital, anesthesia and cardiothoracic surgery departments. PATIENTS: Sixty-two patients without valvular or concomitant diseases undergoing CABG. INTERVENTIONS: CS fluoroscopy, measurements of CS flow, CS oxygen saturation, and CS distal tip pressure before incision, after incision, 20 min after aortic cross clamp release (X-off), 50 min after X-off, 2 h after X-off, 4 h after X-off, and 6 h after X-off. RESULTS: In 57 patients (92%), we achieved successful CS catheter placement. In five patients (8%), CS catheter positioning was not possible. Of the 57 CS catheters placed, dislocation occurred during the operation in six patients (11%) and postoperatively in three patients (6%). Cardiac complications of CS catheter placement occurred in nine patients (15%). Four patients (6%) acquired hemopericardium. Three of these patients had a small hematoma in the right ventricle. In two other patients, contrast medium appeared in the right ventricular wall during catheterization. No hemodynamic signs of these complications were detected clinically. Irregular heart rhythm was observed in only three patients. CS blood oxygen saturation ranged from 40 to 60%. CS flow amounted to 3% of cardiac output. Variations in CS flow paralleled changes in cardiac output. CONCLUSIONS: A CS catheter is a useful tool for clinical human cardiac research; however, the placement of a CS catheter can cause minor myocardial damage in > 10% of patients. Importantly, this damage may not be clinically evident, but only observed after thoracotomy. CS oxygen saturation, CS flow, distal tip pressure, and fluoroscopy are reliable tools to assess a safe and correct positioning of the CS catheter. PMID- 14555555 TI - Relative coronary flow velocity reserve improves correlation with stress myocardial perfusion imaging in assessment of coronary artery stenoses. AB - STUDY OBJECTIVE: To evaluate the angiographic and coronary flow velocity parameters that best correlate with the results of stress myocardial perfusion imaging. DESIGN: Criterion standard. SETTING: Tertiary care center. PATIENTS: Forty-eight patients undergoing diagnostic coronary angiography for angina or silent ischemia. INTERVENTIONS: We performed angiographic and coronary flow velocity measurements at rest and during hyperemia at the post-stenotic segment and in the adjacent angiographically normal branch of the left coronary artery. Relative coronary flow velocity reserve (RCFVR) was calculated as the ratio of post-stenotic to reference vessel coronary flow velocity reserve (CFVR). The best cutoff points for reversible perfusion defects were calculated using receiver operating characteristic curves. MEASUREMENTS AND RESULTS: Post-stenotic CFVR showed fairly good correlations with minimal lumen diameter and percentage of diameter stenosis (r = 0.57 and r = 0.55, respectively; p < 0.001). RCFVR showed stronger correlations with these angiographic indexes of stenosis severity (r = 0.66 and r = 0.68, respectively; p < 0.0001). Based on receiver operating characteristic cutoff values (1.67 for post-stenotic CFVR and 0.64 for RCFVR), RCFVR had better agreement with myocardial perfusion imaging results, compared to post-stenotic CFVR (92% vs 75%, respectively). This agreement was more meaningful in patients with moderate coronary artery stenoses (50 to 75%). The area under the curve was 0.65 (not significant) for post-stenotic CFVR and 0.88 (p < 0.01) for RCFVR. CONCLUSIONS: RCFVR describes better than post-stenotic CFVR the functional significance of coronary artery stenoses. PMID- 14555556 TI - Diagnostic usefulness of carotid pulse tracing in patients with hypertrophic obstructive cardiomyopathy due to midventricular obstruction: a comparison with idiopathic hypertrophic subaortic stenosis. AB - STUDY OBJECTIVES: Of the hypertrophic obstructive cardiomyopathies, midventricular obstruction (MVO) often has been overlooked. In this study, hemodynamic parameters in patients with MVO were compared with patients with idiopathic hypertrophic subaortic stenosis (IHSS), following which the specific markers for diagnosis of MVO were examined. PATIENTS AND DESIGN: Twenty healthy control subjects (mean [+/- SD] age, 54 +/- 8 years), 20 patients with MVO (mean age, 54 +/- 13 years), and 12 patients with IHSS (mean age, 58 +/- 12 years) participated in this study. Hemodynamic parameters associated with carotid pulse tracing (CPT) and echocardiography were examined. MEASUREMENTS AND RESULTS: Left ventricular ejection time (LVET) and left ventricular pressure gradient (LVPG) were greater in patients with IHSS than in patients with MVO (p < 0.0001 for both). However, left ventricular dimensions and interventricular septal thickness did not vary between patients with MVO and those with IHSS. As specific markers for the diagnosis of patients with MVO, two specific CPT patterns, the "spike-and dip pattern" and the "spike-and-half-dome pattern," were identified, but no specific markers were observed echocardiographically. Among patients with MVO, both LVPG and LVET were greater in patients with the spike-and-half-dome pattern than in patients with the spike-and-dip pattern (113 +/- 34 vs 57 +/- 17 mm Hg, respectively [p < 0.0001]; 318 +/- 19 vs 281 +/- 27 ms, respectively [p = 0.0033]), but echocardiographic parameters revealed no significant differences between the two types of MVOs. The pattern of continuous-wave Doppler recordings of the left ventricle in patients with the spike-and-half-dome pattern was identical to that of patients with IHSS, but that of patients with the spike-and dip pattern exhibited concavity from the onset of systole to the point of maximal velocity. CONCLUSIONS: Two specific patterns for the diagnosis of patients with MVO were identified by CPT. These patterns may be strongly related to differences in ejection dynamics. PMID- 14555557 TI - Intracoronary beta-irradiation with liquid rhenium-188: results of the Taiwan radiation in prevention of post-pure balloon angioplasty restenosis study. AB - STUDY OBJECTIVE: To assess the feasibility and short-term outcome of intracoronary irradiation after pure balloon angioplasty (POBA) of de novo and post-POBA restenotic lesions with a liquid beta-emitter (188)Re-filled balloon. DESIGN AND SETTING: Nonrandomized prospective study with contemporaneous control group in a single medical center. PATIENTS AND METHODS: In the Taiwan Radiation in Prevention of Post-Pure Balloon Angioplasty Restenosis study, 40 patients underwent 14-Gy irradiation and 15 patients underwent 20-Gy irradiation at a tissue depth of 0.5 mm after POBA. Thirty control patients received a 5-min inflation with a perfusion balloon catheter after POBA. RESULTS: No procedural or in-hospital complications, or 30-day major adverse cardiac events were noted. Six month angiographic restenosis rates were 49% in the 14-Gy group, 20% in the 20-Gy group, and 57% in the control group (p = 0.05, 20-Gy group vs control group). In the lesions with an arc of calcification of < 180 degrees, restenosis occurred in 15 of the 34 lesions (44%) in the 14-Gy group and in none of the 11 lesions (0%) in the 20-Gy group (p = 0.007). In a vessel with a reference diameter < 3.0 mm, restenosis occurred in 1 of the 8 lesions (13%) in the 20-Gy group, and in 8 of the 11 lesions (73%) in the control group (p = 0.02). In the post-POBA restenotic lesions, restenosis occurred in none of the six lesions (0%) in the 20-Gy group, and in five of the six lesions (83%) in the control group (p = 0.008). CONCLUSIONS: Post-POBA, catheter-based brachytherapy in nonstented native coronary artery with a (188)Re-filled balloon can effectively reduce target lesion restenosis with 20-Gy irradiation at a tissue depth of 0.5 mm and seems to be more effective in the treatment of lesions with an arc of calcification < 180 degrees, in a vessel with a reference diameter of < 3.0 mm, and in post-POBA restenotic lesions. PMID- 14555558 TI - Pharmacodynamics and pharmacokinetics of inhaled iloprost, aerosolized by three different devices, in severe pulmonary hypertension. AB - BACKGROUND: Inhalation of iloprost, a stable prostacyclin analog, is an effective therapy for pulmonary hypertension with few side effects. This approach may, however, be handicapped by limitations of currently available nebulization devices. We assessed whether the physical characterization of a device is sufficient to predict drug deposition and pharmacologic effects. METHODS: We investigated the effects of a standardized iloprost aerosol dose (5 micro g; inhaled within approximately 10 min) in 12 patients with severe pulmonary hypertension in a crossover design employing three well-characterized nebulizers. The nebulizers use different techniques to increase efficiency and alveolar targeting (Ilo-Neb/Aerotrap [Nebu-Tec; Elsenfeld, Germany], Ventstream [MedicAid; Bognor Regis, UK], and HaloLite [Profile Therapeutics; Bognor Regis, UK]). Measurements were performed using a Swan-Ganz catheter and determination of arterial iloprost plasma levels. RESULTS: During inhalation of iloprost, the pulmonary vascular resistance decreased substantially (baseline, approximately 1,250 dyne.s.cm(-5); decrease, - 35.5 to - 38.0%) and pulmonary artery pressure decreased substantially (baseline, approximately 58 mm Hg; decline, - 18.4 to 21.8%), whereas the systemic arterial pressure was largely unaffected. Cardiac output and mixed venous and arterial oxygen saturation displayed a marked increase. The pharmacodynamic profiles with the three devices were superimposable. Moreover, rapid entry of iloprost into the systemic circulation was noted, peaking immediately after termination of the inhalation maneuver, with very similar maximum serum concentrations (158 pg/mL, 155 pg/mL, and 157 pg/mL), and half-lives of serum levels (6.5 min, 9.4 min, and 7.7 min) for the three nebulizers, respectively. Interestingly, the "half-life" of the pharmacodynamic effects in the pulmonary vasculature (eg, decrease in pulmonary vascular resistance, ranging between 21 and 25 min) clearly outlasted this serum level based pharmacokinetic half-life. CONCLUSIONS: A standardized dose of aerosolized iloprost delivered by different nebulizer types induces comparable pharmacodynamic and pharmacokinetic responses. Pulmonary vasodilation, persisting after disappearance of the drug from the systemic circulation, supports the hypothesis that local drug deposition largely contributes to the preferential pulmonary vasodilation in response to inhaled iloprost. PMID- 14555559 TI - Increase in thrombomodulin concentrations after pulmonary thromboendarterectomy in chronic thromboembolic pulmonary hypertension. AB - STUDY OBJECTIVES: The objectives of the study were as follows: (1) to identify differences in endothelial dysfunction and altered hemostasis in patients with chronic thromboembolic pulmonary hypertension (CTEPH) compared with patients with acute pulmonary thromboembolism (APTE) uncomplicated by pulmonary arterial hypertension, by measuring the concentrations of thrombomodulin (TM), a receptor for thrombin and a major anticoagulant proteoglycan on the endothelial membrane, and other plasma factors of coagulation and fibrinolysis; and (2) to examine the effects of thromboendarterectomy on TM levels as a parameter of endothelial cell injury leading to abnormal hemostasis as well as to examine the clinical significance of TM as a marker of endothelial injury. DESIGN: Prospective comparison of concentrations of TM and other plasma parameters among patients with CTEPH or APTE and control subjects. PARTICIPANTS: We studied 22 healthy subjects (ie, control subjects), 22 patients who had been clinically stabilized after APTE, and 44 patients with CTEPH. In 21 of the patients with CTEPH, measurements were repeated after they had undergone pulmonary thromboendarterectomy. MEASUREMENTS AND RESULTS: Plasma concentrations of soluble TM in patients with CTEPH were measured and compared with those in patients with APTE. The mean (+/- SD) TM concentration in the CTEPH group (2.5 +/- 0.7 ng/mL) was significantly lower than that in the control group (4.0 +/- 0.6 ng/mL; p < 0.05). In contrast, the mean plasma TM concentration in the APTE group (4.6 +/- 1.9 ng/mL) was similar to that in the control group. After patients underwent pulmonary thromboendarterectomy, the mean TM concentration increased from 2.0 +/- 0.4 to 2.9 +/- 0.7 ng/mL (p < 0.05). In the CTEPH group, the plasma TM concentration was negatively correlated with mean pulmonary arterial pressure and total pulmonary resistance (p < 0.05). CONCLUSIONS: A decreased plasma TM concentration may reflect pulmonary vascular endothelial dysfunction leading to altered anticoagulant and fibrinolytic function in CTEPH, which rarely develops after APTE. Plasma TM measurements may be useful in distinguishing CTEPH with severe pulmonary hypertension from recurrent APTE. PMID- 14555560 TI - Effects of short-term 28% and 100% oxygen on PaCO2 and peak expiratory flow rate in acute asthma: a randomized trial. AB - STUDY OBJECTIVE: We conducted the first randomized controlled study to assess the effects of short-term 28% and 100% oxygen on PaCO(2) and peak expiratory flow rate (PEFR) in patients with acute severe asthma. PATIENTS AND INTERVENTIONS: Seventy-four patients (mean age, 37.9 +/- 9.7 years [+/- SD]; PEFR, 41.0 +/- 12.1% of predicted) from two emergency departments were randomized to receive 28% or 100% oxygen during 20 min. RESULTS: The administration of 100% oxygen significantly increases PaCO(2) (p = 0.03) and decreases PEFR (p = 0.001) as compared with administration of 28% oxygen. PaCO(2) before and during oxygen administration correlated significantly (p = 0.001) in both groups. Patients breathing 28% oxygen experienced a PaCO(2) fall; on the contrary, patients who received 100% oxygen showed an increase in PaCO(2), particularly those with PaCO(2) before oxygen treatment > 40 mm Hg. CONCLUSIONS: This study confirmed previous observations that oxygen dose should be variable and based on achieving and maintaining target arterial oxygen saturation measured by pulse oximetry > or = 92% rather than on prescribing predetermined concentrations or flow rates of inspired oxygen. PMID- 14555561 TI - A comparison of the clinical characteristics of children and adults with severe asthma. AB - OBJECTIVES: This study sought to better define the clinical characteristics of severe asthma in both children and adults, and to evaluate the effect of asthma duration on multiple parameters of disease severity. DESIGN: Retrospective analysis of prospectively collected data on 275 patients (125 children) with severe asthma who were admitted to a tertiary asthma referral center. METHODS: Demographics, lung function (ie, spirometry and body box plethysmography), glucocorticoid (GC) pharmacokinetic studies, and lymphocyte stimulation assays were performed on all patients. RESULTS: Children were as likely to require therapy with high-dose inhaled GCs and long-term therapy with oral GCs, and to have had a prior intubation, yet they had significantly less airflow limitation (mean [+/- SEM] FEV(1), 74.0 +/- 2.1% predicted vs 57.1 +/- 1.8% predicted, respectively; p < 0.0001), less resistance to airflow (mean airway resistance, 140.3 +/- 8.5% predicted vs 311 +/- 18% predicted, respectively; p < 0.0001), and larger lung volumes (mean total lung capacity, 116.4 +/- 1.6% predicted vs 105.3 +/- 1.8% predicted, respectively; p < 0.0001) compared to adults. Children were more likely to be male and to display greater responsiveness to GCs in vitro. Lung function impairment was associated with asthma duration in children and in adults with onset of asthma in childhood, while there was no relationship between disease severity and asthma duration among those with adult-onset asthma. Despite significant differences in disease duration, patients with adult-onset asthma had equally compromised lung function compared to adults with long-standing asthma. CONCLUSIONS: Children with severe asthma tended to be male, to have less severe airflow obstruction, and to display greater responsiveness to GCs in vitro compared to adults. Symptoms and episodic acute declines in lung function may precede chronic airflow limitation in this group of children. As such, it may be more relevant to follow the deterioration in lung function over time in children. Finally, disease severity in children and adults whose onset of asthma occurred in childhood was related to disease duration, but not in patients with onset of asthma in adulthood. PMID- 14555562 TI - Airway responsiveness to adenosine 5'-monophosphate and exhaled nitric oxide measurements: predictive value as markers for reducing the dose of inhaled corticosteroids in asthmatic subjects. AB - OBJECTIVES: To investigate the utility of the determination of airway responsiveness to inhaled adenosine 5'-monophosphate (AMP) and exhaled nitric oxide (ENO) levels as markers for safely reducing the dose of inhaled corticosteroids (ICS) in patients with asthma well controlled with a moderately high ICS dose. METHODS: A total of 37 patients with asthma well controlled for at least 3 months by treatment with a moderately high ICS dose (beclomethasone dipropionate, 500 to 1,000 microg or equivalent daily) were included in the study. Patients were treated for a 2-week run-in (baseline) period with their usual dose of ICS. For the next 12 weeks, patients were treated with ICS at half the previous dose, maintaining the same inhalation device. At the end of the baseline period and after 2 weeks, 8 weeks, and 12 weeks of treatment with a reduced dose of ICS, measurements were made in the following order: ENO, spirometry, and AMP challenge. Furthermore, patients completed a diary twice daily recording peak expiratory flow, daytime and nighttime symptoms, and use of rescue albuterol. RESULTS: Ten patients had an asthma exacerbation. Using a Kaplan-Meier survival analysis, the significant predictors of a failure of ICS reduction were having both bronchoconstriction in response to AMP and ENO levels > or = 15 parts per billion (ppb) at baseline (p = 0.006), as well as having both bronchoconstriction in response to AMP and ENO levels > or = 20 ppb at baseline (p = 0.033). Having a decrease in the provocative concentration of AMP causing a 20% fall in FEV(1) of at least one doubling concentration 2 weeks after the dose of ICS was halved was a borderline significant predictor for failure of ICS reduction (p = 0.062). CONCLUSION: These observations suggest that in asthmatic patients well controlled with ICS, the determination of AMP responsiveness and ENO levels may be useful to identifying those subjects whose condition will or will not deteriorate when the dose of ICS is reduced. PMID- 14555563 TI - Effect of montelukast on exhaled nitric oxide and nonvolatile markers of inflammation in mild asthma. AB - STUDY OBJECTIVES: Leukotriene receptor antagonists appear to exert anti inflammatory activity in asthma. We undertook the present study to evaluate the effect of montelukast on levels of exhaled nitric oxide (ENO) and two inflammatory markers, hydrogen peroxide (H(2)O(2)), and cysteinyl leukotrienes (cys-LTs), in the exhaled breath condensate of subjects with mild asthma. PATIENTS: Twenty stable subjects with mild asthma (15 women and 5 men; mean [+/- SD] age, 34.8 +/- 12.6 years) were included in the study. INTERVENTION: A 1-week run-in period was followed by 2 weeks of treatment (with montelukast or placebo) that was administered in randomized, double-blind, crossover fashion. One week of washout followed each treatment arm. RESULTS: Montelukast significantly reduced the levels of ENO from baseline (median, 52.5 parts per billion [ppb]; 25th to 75th percentile, 37.8 to 101.8 ppb) during the entire treatment period (ie, day 1 to day 14), with the effect measurable as early as day 1 (median, 45.9 ppb; 25th to 75th percentile, 29.3 to 92.5 ppb) and with the maximal effect being observed on day 7 (median, 35.7 ppb; 25th to 75th percentile, 27.6 to 66.6 ppb). The levels of ENO did not change significantly with placebo therapy. Montelukast improved symptom score and reduced peak expiratory flow (PEF) variability. Changes in PEF variability correlated positively with changes in ENO (r = 0.46; p = 0.04). No significant changes in FEV(1) or concentration of H(2)O(2) in the exhaled breath condensate were observed. Levels of cys-LTs were undetectable in the exhaled breath condensate. CONCLUSIONS: We concluded that montelukast reduces the levels of ENO in patients with mild asthma, a finding that is compatible with an anti-inflammatory effect of montelukast, and that ENO appears to be more sensitive in detecting this effect than FEV(1) and H(2)O(2) levels in the exhaled breath condensate. PMID- 14555564 TI - Sensitivity and validity of three bronchial provocation tests to demonstrate the effect of inhaled corticosteroids in asthma. AB - STUDY OBJECTIVES: To compare the sensitivity and validity of mannitol, histamine, and cold air challenges to demonstrate the effect of inhaled corticosteroids (ICS) in asthma. DESIGN: A prospective study. PARTICIPANTS: Seventeen patients with recently diagnosed, steroid-naive asthma who fulfilled the diagnostic criteria of Finnish Social Insurance Institution and were hyperresponsive to both mannitol and histamine. INTERVENTIONS: The following procedures were carried out at baseline and after 3 months and 6 months of treatment with inhaled budesonide, 800 microg/d: symptom assessment with a questionnaire, ambulatory peak expiratory flow (PEF) measurements twice daily for 2 weeks, and bronchial challenges with mannitol, histamine, and cold air. RESULTS: Budesonide decreased the sum symptom score, daily use of bronchodilating drugs, and diurnal PEF variation, but did not change FEV(1) percentage of predicted significantly. In addition, budesonide significantly decreased mannitol (p = 0.005) and histamine (p = 0.002) response dose ratios. The magnitude of the budesonide-induced change in responsiveness to these two challenges did not differ significantly. The effect of budesonide on cold air responsiveness did not reach statistical significance (p = 0.064). Change in mannitol responsiveness correlated significantly with the changes in sum symptom score and in FEV(1). Change in cold air responsiveness correlated with the changes in sum symptom score and in diurnal PEF variation. Change in histamine responsiveness correlated only with change in FEV(1). CONCLUSIONS: Mannitol challenge is both a sensitive and valid test to demonstrate the effects of ICS in asthma. Histamine challenge is equally sensitive for this purpose, but its validity may be lower than that of mannitol challenge. Cold air challenge seems to be a valid test to demonstrate the effects of ICS, but its sensitivity may be lower than that of mannitol and histamine challenges. PMID- 14555565 TI - Withdrawal from treatment as an outcome in the ISOLDE study of COPD. AB - OBJECTIVES: To investigate the determinants of patient withdrawal from our study, and the effect of these withdrawals on the outcome of treatment with inhaled corticosteroids in patients with COPD. DESIGN: A double-blind, placebo controlled, randomized trial. SETTING: Eighteen outpatient centers in the United Kingdom. PARTICIPANTS: Seven hundred fifty-one patients with stable COPD defined clinically and as baseline postbronchodilator FEV(1) > or = 0.8 L and < 85% predicted, FEV(1)/FVC ratio < 70%, and FEV(1) change after albuterol < 10% of predicted. INTERVENTION: Random assignment of either 500 microg bid of inhaled fluticasone propionate (FP) using a spacer device or an identical placebo inhaler. Treatment was continued for 3 years or until patients withdrew from follow-up. MEASUREMENTS AND RESULTS: Postbronchodilator FEV(1) was measured on three occasions before randomization and every 3 months thereafter. Health status was assessed by the disease-specific St. George Respiratory Questionnaire (SGRQ) and the modified short-form 36 questionnaire (SF-36) at baseline and every 6 months. Three hundred thirty-nine patients withdrew, of whom 156 patients received FP. Prescription of frequent courses of oral prednisolone was the most common reason for withdrawing as specified in the protocol (69 patients in the FP group withdrew due to respiratory symptoms, compared with 93 patients in the placebo group). This explained the significantly greater dropout of placebo treated patients that was most evident when FEV(1) was < 50% predicted. Patients withdrawing had a significantly more rapid decline in health status, measured by both the SGRQ and the SF-36 (p < 0.001). Those withdrawing from the placebo group had a more rapid decline in FEV(1) and more exacerbations than the FP-treated groups. Baseline FEV(1) was lower in dropouts than in patients completing the study receiving placebo, but there was no difference between the respective groups receiving FP. CONCLUSIONS: Patients who withdrew from follow-up were those with the most rapidly deteriorating health status and lung function. Losing these patients from the final analysis can reduce the power of a study to achieve its primary end point. PMID- 14555566 TI - Comparison of specific expiratory, inspiratory, and combined muscle training programs in COPD. AB - BACKGROUND: Respiratory muscle weakness may contribute to dyspnea and exercise limitation in patients with significant COPD. In an attempt to reduce the severity of breathlessness and to improve exercise tolerance, inspiratory muscle training has been applied in many COPD patients. On the other hand, there is a paucity of data related to expiratory muscle performance and training in COPD. METHODS: Thirty-two patients with significant COPD (ie, mean FEV(1), 37% of predicted) were recruited for the study. The patients were randomized into four groups: eight patients were assigned to receive specific expiratory muscle training (SEMT); eight patients received specific inspiratory muscle training (SIMT); eight patients received SEMT and SIMT (ie, the SEMT + SIMT group); and eight patients who were assigned to a control group received training with very low load. All patients trained daily, six times a week, with each session consisting of one half hour of training, for 3 months. Spirometry, respiratory muscle strength and endurance, 6-min walk test distance, the perception of dyspnea, and the Mahler baseline dyspnea index (BDI) were measured before and following training. RESULTS: Training caused a statistically significant specific increase in the expiratory muscle strength and endurance (in the SEMT and SEMT + SIMT groups) and in the inspiratory muscle strength and endurance (in the SIMT and SEMT + SIMT groups). There was significant increase in the distance walked in 6 min in the SEMT, SIMT, and SEMT + SIMT groups. However, the increase in the SIMT and SEMT + SIMT groups was significantly greater than that in the SEMT group. There was a statistically significant increase in the BDI, and a decrease in the mean Borg score during breathing against resistance in the SIMT and SEMT + SIMT groups, with no changes in the SEMT and control groups. CONCLUSIONS: The inspiratory and expiratory muscles can be specifically trained with improvement of both muscle strength and endurance. The improvement in the inspiratory muscle performance is associated with an increase in the 6-min walk test distance and the sensation of dyspnea. There is no additional benefit gained by combining SIMT with SEMT, compared to using SIMT alone. PMID- 14555567 TI - Body composition and resting energy expenditure in clinically stable, non-weight losing patients with severe emphysema. AB - STUDY OBJECTIVES: To characterize the metabolic status of weight-stable and clinically stable individuals with advanced emphysema. PATIENTS: Seventy-nine patients with severe emphysema (FEV(1), 29 +/- 13% of predicted [mean +/- SD]) evaluated for enrollment in the National Emphysema Treatment Trial and 20 age matched healthy subjects were studied. SETTING: Pulmonary function laboratory of university-affiliated teaching hospital. INTERVENTIONS: Data collection. MEASUREMENTS AND RESULTS: We measured lung function, body composition, serum leptin levels, serum tumor necrosis factor receptors (sTNF-Rs), resting oxygen consumption (RVO(2)) normalized to weight in kilograms (RVO(2)/kg), and RVO(2) normalized to fat-free mass (FFM) [RVO(2)/FFM]. The patient group and healthy group had similar age, body mass index (BMI), and body composition. RVO(2)/kg, RVO(2)/FFM, and sTNF-R levels were higher in patients compared to healthy subjects. There were no differences in serum leptin levels between emphysematous and healthy subjects, and there was no correlation between leptin and sTNF-R and RVO(2)/kg. Furthermore, both groups had similar gender-related differences in FFM, percentage of body fat, and serum leptin levels. Patients with lower BMI showed the greatest differences from control subjects in RVO(2)/kg. CONCLUSION: In weight-stable subjects with advanced emphysema, RVO(2)/kg and RVO(2)/FFM were higher compared to healthy subjects, especially in those with BMI in the lower end of the normal range. RVO(2)/kg and RVO(2)/FFM did not correlate with leptin or sTNF-R levels. These data show that a higher metabolic rate is found in patients with emphysema who are clinically and weight stable. Thus, hypermetabolism is a feature of the disease and not sufficient to lead to weight loss. PMID- 14555568 TI - Oxidative stress in expired breath condensate of patients with COPD. AB - OBJECTIVE: To evaluate the levels of hydrogen peroxide (H(2)O(2)) and 8 isoprostane in the expired breath condensate (EBC) of patients with COPD, and to assess the relationship between the above markers of oxidative stress and parameters expressing inflammatory process and disease severity. SETTING: Inpatient respiratory unit and outpatient clinic in tertiary care hospital. DESIGN: Cross-sectional study. PATIENTS: Thirty stable COPD patients (all smokers) with disease severity ranging from mild to severe. Ten subjects who were smokers with stage 0 disease (ie, at risk for COPD; mean [+/- SD] FEV(1), 88 +/- 5% predicted) were studied as a control group. METHODS: H(2)O(2) and 8 isoprostane levels were measured in EBC, and the values were correlated with variables expressing COPD severity (ie, FEV(1) percent predicted, dyspnea severity score (ie, Medical Research Council scale) and airway inflammation (ie, differential cell counts from induced sputum). RESULTS: The mean concentration of H(2)O(2) was significantly elevated in COPD patients compared to control subjects (mean, 0.66 micromol/L [95% confidence interval (CI), 0.54 to 0.68 micro mol/L) vs 0.31 micro mol/L [95% CI, 0.26 to 0.35 micromol/L], respectively; p < 0.0001). The difference was primarily due to the elevation of H(2)O(2) in patients with severe and moderate COPD, whose expired breath H(2)O(2) levels were significantly higher than those of patients with mild disease (mean, 0.96 micromol/L [95% CI, 0.79 to 1.13 micromol/L], 0.68 micromol/L [95% CI, 0.55 to 0.81 micromol/L], and 0.33 micromol/L [95% CI, 0.24 to 0.43 micromol/L], respectively, p < 0.0001). The mean concentration of 8-isoprostane was significantly elevated in patients with COPD compared to that of the control group (47 pg/mL [95% CI, 41 to 53 pg/mL] vs 29 pg/mL [95% CI, 25 to 33 pg/mL], respectively; p < 0.0001) but did not differ significantly among the different stages of the disease (p = 0.43). Repeatability and stability data within measurements showed that H(2)O(2) has a better repeatability and stability than 8-isoprostane. Furthermore, we observed significant correlations of H(2)O(2) with FEV(1), neutrophil count, and dyspnea score. Those correlations existed only in patients with moderate and severe disease. No correlations were found between levels of 8-isoprostane and the above parameters. CONCLUSIONS: We conclude that levels of H(2)O(2) and 8-isoprostane are elevated in the EBC of patients with COPD, but that H(2)O(2) seems to be a more repeatable and a more sensitive index of the inflammatory process and the severity of the disease. PMID- 14555569 TI - Plasma orexin-A levels in obstructive sleep apnea-hypopnea syndrome. AB - STUDY OBJECTIVES: Orexin and orexin receptors are present in the CNS. The effects of orexin peptides have been uniformly reported as excitatory, and the posterior hypothalamus containing orexin neurons has been implicated in arousal state control. Therefore, it is probable that the orexin system may have a neuromodulatory effect on arousal states. The aim of the present study was to investigate the relationship between plasma orexin-A levels and arousals from sleep in patients with obstructive sleep apnea-hypopnea syndrome (OSAHS). DESIGN: An analysis was conducted in 30 male patients with OSAHS, which had been diagnosed by polysomnography by the presence of an apnea-hypopnea index (AHI) of > 5, and 20 male age-matched and body mass index (BMI)-matched control subjects. RESULTS: Plasma orexin-A levels were higher in patients with OSAHS compared with those in control subjects (p < 0.05). Plasma orexin-A levels correlated positively, but weakly, with the arousal index (r = 0.51; p < 0.05) and the AHI (r = 0.52; p < 0.05). However, plasma orexin-A levels did not relate to age, BMI, Epworth sleepiness scale, PaO(2), PaCO(2), minimum arterial oxygen saturation (SaO(2)) during sleep, or mean SaO(2) during sleep. Plasma orexin-A levels can be a measure of both AHI and arousal index. CONCLUSION: These results suggested that the orexin system may be involved in arousal mechanisms in patients with OSAHS. PMID- 14555570 TI - 8-Isoprostane, a marker of oxidative stress, is increased in exhaled breath condensate of patients with obstructive sleep apnea after night and is reduced by continuous positive airway pressure therapy. AB - STUDY OBJECTIVES: Obstructive sleep apnea (OSA) is characterized by recurrent apnea during sleep that may compromise oxidative balance. Oxidative stress is increased in the blood and in the airways of OSA patients. DESIGN: The aim of this study was to investigate whether oxidative stress is determined by nocturnal apneas and could be reduced by CPAP therapy, and whether there is a relation between local and systemic oxidative stress in these patients. PATIENTS AND METHODS: Eighteen patients with OSA (13 men; mean [+/- SD] age, 48 +/- 3 years) and 12 healthy age-matched and weight-matched subjects (8 men; mean age, 46 +/- 7 years) were recruited. 8-Isoprostane was measured in exhaled breath condensate and blood by a specific enzyme immunoassay. MEASUREMENTS AND RESULTS: Higher concentrations of 8-isoprostane were found in the morning exhaled condensate (9.5 +/- 1.9 pg/mL) and plasma (9.7 +/- 1.5 pg/mL) of OSA patients compared to healthy obese subjects (6.7 +/- 0.2 and 7.1 +/- 0.3 pg/mL, respectively; p < 0.0001). Elevated mean concentrations of exhaled 8-isoprostane were observed in the OSA patients at 8:00 AM (9.5 +/- 1.9 pg/mL) but not at 8:00 PM (7.6 +/- 0.8 pg/mL; p < 0.0005), and a significant reduction was seen after continuous positive airway pressure (CPAP) therapy (7.7 +/- 0.9 pg/mL; before treatment, 9.6 +/- 1.7 pg/mL; p < 0.005). A positive correlation was found between morning exhaled 8 isoprostane levels and the apnea-hypopnea index (r = 0.8; p < 0.0001), and 8 isoprostane levels and neck circumference (r = 0.6; p < 0.0001). CONCLUSIONS: These findings suggest that systemic and local oxidative stress are increased in OSA patients, and that they are higher after nocturnal apnea and reduced by CPAP therapy. PMID- 14555571 TI - 24-hour BP in children with congenital central hypoventilation syndrome. AB - OBJECTIVE: To study circadian BP patterns in patients with congenital central hypoventilation syndrome (CCHS). DESIGN: Case-control study. SETTING: Teaching hospital in Paris, France. PATIENTS: Eleven patients with CCHS (median age, 13 years; range, 6 to 18 years) and 11 sex- and height-matched control subjects. INTERVENTION: None. METHODS: Each subject underwent 24-h ambulatory BP monitoring. Oxygen saturation and end-tidal PCO(2) were monitored noninvasively. Polysomnography was performed to determine sleep times. All patients with CCHS received mechanical ventilation during sleep. Mean values for systolic BP (SBP) and diastolic BP (DBP) during wakefulness and sleep were analyzed. Nocturnal BP "dipping" was defined as the difference in mean SBP (and/or DBP) between wakefulness and sleep, divided by individual waking mean values. BP "dippers" were defined as subjects showing at least 10% nocturnal dipping. RESULTS: Patients with CCHS had BPs in the low normal range of normative data. As compared to control subjects, patients with CCHS had lower BP during wakefulness (p = 0.003 and p = 0.016 for SBP and DBP, respectively), and higher BP during sleep (p = 0.016 and p = 0.002). Nocturnal BP dipping was abnormally reduced in patients with CCHS (p = 0.000). Ten of the 11 patients with CCHS were BP nondippers, compared to none of the control subjects. CONCLUSION: The abnormal circadian BP pattern observed in children and adolescents with CCHS may be related to autonomic nervous dysfunction. Lifelong cardiovascular follow-up is recommended for patients with CCHS. PMID- 14555572 TI - The relationship between congestive heart failure, sleep apnea, and mortality in older men. AB - STUDY OBJECTIVES: To examine the association of sleep apnea with heart disease. DESIGN: Prospective study. SETTING: Medical wards at the Veterans Affairs San Diego Healthcare System. PATIENTS: Three hundred fifty-three randomly selected inpatient men. MEASUREMENTS AND RESULTS: Sleep was recorded for 2 nights in the hospital. Medical conditions were obtained from hospital medical records. Cox proportional hazards analyses indicated that patients with congestive heart failure (CHF) plus central sleep apnea (CSA) had shorter survival than those with just CHF, just sleep apnea (obstructive or central), or neither. Survival for those with obstructive sleep apnea (OSA) or CSA and no CHF was no different than for those with neither disorder. Follow-up analysis showed that for those with no CHF, neither CSA nor OSA shortened survival (p > 0.80). For those with CHF, having CSA shortened the life span with a hazard ratio of 1.66 (p = 0.012), but having OSA had no effect. Patients with CHF had more severe sleep apnea than those with no heart disease. CONCLUSIONS: This study does not clarify the issues of cause and effect, but does reinforce the strong associations between sleep apnea and heart disease in elderly men. These data suggest that people with coronary disease should be regarded as a risk group for sleep apnea. PMID- 14555573 TI - Prevalence of symptoms and risk of sleep apnea in primary care. AB - BACKGROUND: To obtain prevalence estimates for key symptoms and features that can indicate the presence of obstructive sleep apnea (OSA) in a broad range of primary care settings. DESIGN: Cross-sectional survey. SETTING: Forty offices and clinics in the United States, Germany, and Spain. PARTICIPANTS: Consecutive patients who were > 15 years of age, regardless of the reason for the visit. MEASUREMENTS: We collected demographic information, prevalence of self-reported chronic snoring, sleepiness, obesity (body mass index [BMI] > 30), hypertension, and calculation of OSA risk, and we also compared results between the United States and Europe. RESULTS: There was a 78% return rate for 8,000 surveys (mean age, 51 years; age range, 15 to 98 years; 52% women). One third of participants (32%) had a high pretest probability for OSA, with a higher rate in the United States (35.8% of 3,915 participants) than in Europe (26.3% of 2,308 participants; p < 0.001; age-matched and sex-adjusted odds ratio [OR], 1.37; 95% confidence interval [CI], 1.16 to 1.61). Sleepiness (32.4% vs 11.8%, respectively; p < 0.001) followed by obesity and/or hypertension (44.8% vs 37.1%, respectively; p < 0.01) contributed to the OSA risk difference between participants in the United States and Europe, as frequent snoring and breathing pauses were similarly reported (44%). A high pretest probability for OSA was more often present in men than in women (37.9% vs 27.8%, respectively; p < 0.005; OR, 1.96; CI, 1.59 to 2.88) and in those that were obese (ie, BMI, > or = 30 kg/m(2)), a condition that is generally more common in the US population than in the European population (27.9% vs 17.2%, respectively; p < 0.01). CONCLUSIONS: Primary care physicians in the United States and Europe will encounter a high demand for services to confirm or manage sleep apnea, sleepiness, and obesity. PMID- 14555574 TI - A comparison of smoking habits among medical and nursing students. AB - OBJECTIVE: The approach and credibility of future physicians and nurses as treatment providers for smoking- and tobacco-related diseases may be influenced by their smoking habits. We compared smoking habits among medical and nursing students, and examined whether these habits changed during the course of education for each cohort. METHOD: Over 1,100 medical and nursing students from a university were surveyed in year 2000 using a questionnaire that included the Fagerstrom test for nicotine dependence (FTND). RESULTS: A total of 397 medical students and 126 nursing students completed the survey. Significantly fewer medical students (3.3%) smoked compared to nursing students (13.5%). Also, significantly more nursing students were former smokers (17.8%) than medical students (9.8%). The severity of nicotine dependence, as indicated by the total FTND score as well as scores on five of the six items on the FTND, was significantly lower among medical students compared to nursing students. Smoking or quit rates did not differ across class years in both groups; however, unlike nursing students, time since quitting significantly differed across class years for medical students. Although smoking habits appear to change little during the course of education for both medical and nursing students, many smokers may have quit just prior to entering medical school but not nursing school. CONCLUSIONS: The findings confirm the continuing decline in smoking among medical students in the United States; however, increased efforts to promote tobacco education and intervention among nursing students seem necessary. Nevertheless, both groups appear to have the potential to be credible advisors to patients and public regarding smoking cessation. PMID- 14555575 TI - A comparison of the original chronic respiratory questionnaire with a standardized version. AB - BACKGROUND AND OBJECTIVES: The chronic respiratory questionnaire (CRQ), a widely used measure of health-related quality of life (HRQL) in patients with chronic airflow limitation, includes an individualized dyspnea domain (patients identify five important activities, and report the degree of dyspnea on a 7-point scale). Because the individualized domain is unwieldy in multicenter clinical trials, we developed a standardized version and tested its discriminative and evaluative properties. METHODS: We enrolled 51 patients who completed the standardized and individualized CRQ before starting a respiratory rehabilitation program, and again 3 months later. We calculated both cross-sectional and longitudinal correlations between the two versions and a number of other HRQL instruments, and tested the relative ability of the individualized and standardized versions of the CRQ to detect improvement with rehabilitation. RESULTS: The results of the individualized questions suggested greater dysfunction (lower scores) than did the standardized questions both at baseline (3.18 vs 3.92, p < 0.001) and follow up (4.62 vs 4.84, p = 0.051). The standardized dyspnea domain showed superior discriminative validity. While both techniques detected important, statistically significant improvement with rehabilitation (individualized domain mean change, 1.44; 95% confidence interval [CI], 1.11 to 1.77 [p < 0.001]; standardized domain mean change, 0.92; 95% CI, 0.61 to 1.24 [p < 0.01]), the difference in effect was substantial and statistically significant (mean difference, 0.52; 95% CI, 0.22 to 0.82; p = 0.001). The two versions showed comparable longitudinal validity. CONCLUSIONS: A standardized version of the CRQ dyspnea domain improves the cross sectional validity, maintains longitudinal validity, but reduces the responsiveness. By increasing sample size, investigators can use the more efficient standardized version of the CRQ without compromising validity. PMID- 14555576 TI - The teaching of chest auscultation during primary care training: has anything changed in the 1990s? AB - OBJECTIVE: To survey the teaching time and importance given to cardiopulmonary auscultation during internal medicine (IM) and family practice (FP) residencies, and to compare current practices to those of the early 1990s. DESIGN: A nationwide mail survey of IM and FP program directors (PDs). SETTING: All Accreditation Council for Graduate Medical Education-accredited IM and FP residencies. PARTICIPANTS: A total of 538 of 939 PDs (57.5%). MEASUREMENTS AND MAIN RESULTS: In contrast to the early 1990s, when there had been no significant difference in teaching practices between IM and FP programs, more IM than FP residencies taught cardiopulmonary auscultation in 1999 (cardiac auscultation: IM residencies, 48%; FP residencies, 29.2% [p < 0.001]; pulmonary auscultation: IM residencies, 23.7%; FP residencies, 12.2% [p < 0.001]). Across the decade there also had been a significant increase in the percentage of IM programs offering structured education in chest auscultation (cardiac auscultation increase, 27.1 to 48% [p < 0.001]; pulmonary auscultation increase, 14.1 to 23.7% [p < 0.02]), but no significant changes for FP residencies. IM PDs gave more clinical importance to auscultation and expressed a greater desire for expanded teaching than did their counterparts in FP programs. CONCLUSIONS: This study indicates a significant gain over the last decade in the percentage of IM residencies offering structured teaching of cardiopulmonary auscultation. This same gain did not occur for FP programs. Whether these differences in attitudes and teaching practices will translate into improved auscultatory proficiency of IM trainees will need to be determined. PMID- 14555577 TI - Hetastarch and bleeding complications after coronary artery surgery. AB - STUDY OBJECTIVES: Controversy persists concerning the potential association between intraoperative use of hetastarch (ie, hydroxyethyl starch [HES]) and postoperative bleeding in patients undergoing surgery. To determine whether intraoperative HES use is associated with an increased risk of postoperative bleeding following coronary artery bypass graft (CABG) surgery. DESIGN: Case control study. SETTING: A large academic medical center in the northeastern United States. PARTICIPANTS: A consecutive sample of 238 patients undergoing CABG surgery. MAIN OUTCOME MEASURES: Cases consisted of patients who had received either > or = 3 U packed RBCs, > or = 3 U platelets, > or= 3 U fresh frozen plasma, or any cryoprecipitate within 72 h after undergoing a CABG procedure, or who had undergone surgical revision for bleeding. All other CABG surgery patients served as control subjects. RESULTS: In multivariate models that controlled for a wide variety of demographic and clinical characteristics, we found that, compared to patients who did not receive any HES during surgery, those who received 1 U intraoperative HES had more than twice the risk of a bleeding outcome (odds ratio [OR], 2.32; 95% confidence interval [CI], 1.10 to 4.91), and those who received 2 or 3 U HES had more than four times the risk of postoperative bleeding (OR, 4.57; 95% CI, 1.74 to 12.00). CONCLUSIONS: HES use in patients undergoing CABG surgery may be associated with a significant risk of postoperative bleeding. A double blinded, randomized, controlled trial will be necessary to confirm this finding. PMID- 14555578 TI - Lung volume reduction surgery in Australia and New Zealand. Six years on: registry report. AB - BACKGROUND: Lung volume reduction surgery (LVRS) has been shown to improve lung function, exercise performance, and quality of life in highly selected individuals with severe emphysema; however, major questions regarding the efficacy and long-term outcomes of LVRS still remain unanswered. Pending the results of large randomized controlled trials (RCTs), the Australian and New Zealand LVRS Database was created to audit local clinical practice and patient outcomes. AIMS: To review patient selection, surgical activity, and patient outcomes related to LVRS in Australia and New Zealand. METHODS: Prospective data were voluntarily submitted by hospitals performing LVRS in Australia and New Zealand. Preoperative, surgical, perioperative, and follow-up variables were analyzed. RESULTS: Data were collected from 15 hospitals regarding 529 patients. Mean age (+/- SD) at surgery was 63 +/- 7 years. Preoperatively, FEV(1) was 29 +/ 9% predicted, total lung capacity (TLC) was 138 +/- 20% predicted, residual volume (RV) was 250 +/- 64% predicted, and 6-min walk (6MW) distance was 327 +/- 111 m. There has been a reduction in the overall number of cases and hospitals performing LVRS since 1999. Improvements in lung function following LVRS (ie, FEV(1) increase of 38%, RV decrease of 27%, TLC decrease of 17%) and exercise capacity (ie, 6MW distance increase of 24%) appear to be maintained for approximately 3 years. CONCLUSIONS: LVRS continues to be performed in Australia and New Zealand, predominantly in large tertiary teaching hospitals with similar outcomes to those described in the literature. It remains difficult to capture long-term lung function and survival outcomes in this population. Ongoing audit and RCTs are both required to resolve the confusion that still shrouds this procedure. PMID- 14555579 TI - Surgical treatment of pacemaker and defibrillator lead endocarditis: the impact of electrode lead extraction on outcome. AB - BACKGROUND: Cardiac device (CD) endocarditis is an infrequent but potentially lethal infectious complication of permanent pacemakers or implantable cardioverter-defibrillators (ICDs), and mortality rates of 30 to 35% have been reported. Medical treatment has been suggested for the treatment of CD endocarditis, but there is increasing evidence that surgical treatment is to be preferred as the best approach to achieve eradication of the infection and reduce mortality. OBJECTIVE: To evaluate the following: (1) the clinical and echocardiographic characteristics of patients with pacemaker or ICD endocarditis, (2) the outcome of this population depending on the mode of treatment (medical vs surgical treatment), and (3) the clinical, microbiological, echocardiographic, and therapeutic variables associated with patient outcome. DESIGN: Prospective cohort study. SETTING: Tertiary referral center in Barcelona, Spain. PATIENTS: All consecutive patients with infectious endocarditis (IE) admitted to the study institution between 1990 and 2001 were prospectively evaluated by a multidisciplinary treatment team, and a definite diagnosis of CD endocarditis was established when cases met pathologic or clinical criteria according to the Duke criteria. RESULTS: A total of 31 patients, 25 men and 6 women aged 61 +/- 15 years (mean +/- SD), with pacemaker or ICD endocarditis were identified among 669 consecutive patients (4.6%) with IE. During the study period, a total of 3,768 pacemakers and 460 ICDs were implanted in the study institution. In 22 cases of pacemaker endocarditis, the pacemaker was implanted in our institution, and 9 cases were referred from other institutions (incidences of endocarditis on pacemaker and ICD implanted in our institution of 0.58% and 0.65%, respectively). Medical treatment without removal of the pacing system was initially performed on seven patients; all of them (100%) had relapses of endocarditis, and one patient died. The remaining 24 patients underwent surgical removal of the pacing system; 1 patient had one relapse, 3 patients died after surgical treatment, and the others were successfully cured with no relapses after a mean follow-up of 38 +/- 9 months. Clinical, echocardiographic, microbiological, and therapeutic variables were evaluated in association with prognosis. The only prognostic factor for failure of treatment or mortality was the absence of surgical treatment (p < 0.0001). CONCLUSIONS: Electrode lead endocarditis occurred in < 1% of pacemaker and ICD implants. Conservative treatment without explantation of all hardware failed in all patients, and surgical treatment during antibiotic therapy was effective in eradication of infection but was associated with a 12.5% mortality. The only patient characteristic associated with treatment failure or death was the absence of surgical removal of all infected hardware. Complete extraction of the pacemaker or ICD should be considered as standard therapy for most patients with CD endocarditis. PMID- 14555580 TI - The effect of pregnancy on survival in women with cystic fibrosis. AB - STUDY OBJECTIVES: Patients with cystic fibrosis (CF) are currently living to their fourth decade and are making reproductive decisions. Information concerning the reproductive health of women with CF has been limited to small or single center studies. DESIGN: We conducted a matched parallel-cohort study to assess the impact of pregnancy on the survival of women with CF. PARTICIPANTS: A parallel-cohort study included all women > 12 years of age who were enrolled in the US Cystic Fibrosis Foundation National Patient Registry from 1985 to 1997. MEASUREMENTS AND RESULTS: Six hundred eighty of the 8,136 women in the cohort became pregnant. These 680 women were matched on an index year to 3,327 control women with CF. At the inception of entry into the cohort, women who reported pregnancy were more likely to have had a higher percentage of predicted FEV(1) (67.5% predicted vs 61.7% predicted, respectively; p < 0.001) and a higher weight (52.9 vs 46.4 kg, respectively; p < 0.001). Using Kaplan-Meier survival curves, the 10-year survival rate in pregnant women (77%; 95% confidence interval [CI], 71 to 82%) was higher than in those women who did not become pregnant (58%; 95% CI, 55 to 62%). A separate analysis, matching pregnant patients on FEV(1) percent predicted, age, Pseudomonas aeruginosa colonization, and pancreatic function, obtained similar results. Using Cox proportional hazard modeling to adjust for baseline age, FEV(1) percent predicted, weight, height, and pulmonary exacerbation rate per year, pregnancy was not associated with an increase risk of death. Pregnancy was not harmful in any subgroup including patients with FEV(1) < 40% of predicted or diabetes mellitus. CONCLUSIONS: Women with CF who became pregnant were initially healthier and had better 10-year survival rates than women with CF who did not become pregnant. After adjustment for the initial severity of illness, women who became pregnant did not have a significantly shortened survival. PMID- 14555581 TI - Utility of the breathing reserve index at the anaerobic threshold in determining ventilatory-limited exercise in adult cystic fibrosis patients. AB - OBJECTIVES: Cardiopulmonary exercise testing in cystic fibrosis (CF) patients is useful to assess functional status and prognosis. Using the current interpretation guidelines, the utility of this testing will be limited in those patients who cannot exercise to a near-maximal level. This study investigates the utility of the breathing reserve index at the anaerobic threshold (BRIAT), which is defined as minute ventilation at the anaerobic threshold (AT)/maximum voluntary ventilation (MVV), to distinguish ventilatory-limited (VL) CF patients from nonventilatory-limited (NVL) CF patients. DESIGN: Exercise studies on 53 adult CF patients at baseline clinical status performed from 1993 to 1999 were reviewed, of which 40 met the inclusion criteria. The studies were performed via ramp protocol to the symptom-limited maximum on a cycle ergometer with breath-by breath expired gas analysis. AT was determined noninvasively via the V-Slope method. The patients were classified as VL if they had abnormal spirometry findings, reduced exercise capacity, and a breathing reserve index at maximum exercise (BRImax) of > or = 0.7. NVL patients had a normal BRImax and met the criteria for a maximal study. RESULTS: VL patients (21 patients) had significantly lower FVC, FEV(1), MVV, and body mass index than NVL patients (19 patients). The BRIAT for the VL group was significantly higher than that for the NVL group (p < 0.001). Logistic regression analysis revealed that BRIAT discriminated VL patients from NVL patients better than a variety of nonexercise variables tested. The BRIAT correlated extremely well with BRImax (r = 0.89; p < 0.01), FVC (r = -0.67; p < 0.001), FEV(1) (r = -0.76; p < 0.001), and FEV(1)/FVC ratio (r = -0.683; p < 0.001). A BRIAT value of 0.29 distinguished VL CF patients from NVL CF patients with 95.2% sensitivity and 84.2% specificity. CONCLUSIONS: The BRIAT assessed noninvasively correlates well with commonly used measurements of pulmonary function and accurately distinguishes CF patients with and without a ventilatory limitation to exercise. The BRIAT may have utility in the interpretation of exercise studies in CF patients who are unable to exercise to a maximal level. PMID- 14555582 TI - Comparative roles of levofloxacin and ofloxacin in the treatment of multidrug resistant tuberculosis: preliminary results of a retrospective study from Hong Kong. AB - OBJECTIVE: To compare levofloxacin and ofloxacin in the treatment of multidrug resistant tuberculosis (MDR-TB). PATIENTS AND METHODS: A retrospective analysis of 106 patients with MDR-TB (February 1990 through December 2000) receiving directly observed therapy with fluoroquinolone and accompanying drugs, which mainly included aminoglycosides, cycloserine, ethionamide/prothionamide, and pyrazinamide, was performed. Clinical data from 99 suitable patients were subjected to univariate analysis, stratification, and multiple logistic regression to compare the roles of levofloxacin and ofloxacin in multidrug regimens. RESULTS: Forty patients received 612.5 +/- 79.0 mg qd levofloxacin (mean +/- SD), and 59 patients received 628.8 +/- 101.8 mg qd ofloxacin together with similar active second-line drugs for similar durations. The times to sputum smear (both 1.8 months) and culture conversion (both 2.1 months) were equivalent. Adverse reactions occurred at similar rates (10.0% vs 11.9%). The combined treatment success rate was 83.8%, being higher among ofloxacin-susceptible than ofloxacin-resistant cases (90.5% vs 64.0%, p < 0.01). The success rates for the levofloxacin group were 90.0% (overall), 96.2% (ofloxacin-susceptible cases), and 78.6% (ofloxacin-resistant cases) in comparison with 79.7%, 87.5%, and 45.5%, respectively, for the ofloxacin group (Mantel-Haenszel common odds ratio estimate, 4.0; p < 0.05). Bacillary susceptibility to ofloxacin, good adherence, radiographic extent of one lung or less, and use of levofloxacin were independent predictors of favorable outcome (odds ratios, 7.6 to 21.3). One patient each from both groups relapsed. CONCLUSION: Levofloxacin was found to be more efficacious than ofloxacin when incorporated into multidrug regimens used for treatment of MDR-TB. PMID- 14555583 TI - Treatment of Mycobacterium avium-intracellulare complex lung disease with a macrolide, ethambutol, and clofazimine. AB - BACKGROUND: Mycobacterium avium-intracellulare (MAC) causes progressive lung disease. Recommended treatment regimens include a macrolide and a rifamycin, but drug intolerance and relapse after treatment is completed often limit successful therapy. METHODS: Consecutive individuals referred for treatment of MAC lung disease were treated with a regimen that included either clarithromycin, 500 mg bid, or azithromycin, 250 mg/d, on weekdays; ethambutol, 15 mg/kg/d; and clofazimine, 100 mg/d. The intention was to treat patients for a minimum of 12 months. The diagnosis of MAC lung disease was confirmed by multiple positive sputum culture findings in patients with typical symptoms and radiologic findings. RESULTS: Thirty patients (27 women and 3 men; mean age, 70 +/- 9.4 years [SD]) were treated. A total of 22 of the patients reported adverse effects from clarithromycin or azithromycin. Intolerance of clarithromycin resulted in the withdrawal of four patients before sputum conversion. The remaining patients continued treatment for an average of 10 months, and sputum findings converted to negative in all 26 patients (87%). One patient died of unrelated causes while still receiving therapy, and five patients (19%) relapsed an average of 17 months after treatment was completed. CONCLUSIONS: Treatment with a macrolide, ethambutol, and clofazimine was successful in 20 of 30 patients (67%) with MAC lung disease and is a reasonable alternative to rifamycin-containing regimens. PMID- 14555584 TI - Pulse transit time as a measure of inspiratory effort in children. AB - STUDY OBJECTIVES: The current criterion standard for measuring inspiratory effort, esophageal manometry, is an invasive procedure that young patients find intolerable. Inspiratory effort can also be assessed noninvasively by measuring the pulse transit time (PTT). PTT is the time the pulse wave (PW) takes to travel between two arterial sites (normally heart to finger). The speed at which the PW travels is directly proportional to arterial BP. When BP rises, PTT shortens. Conversely, when BP falls, PTT lengthens. In this study, we investigated PTT as a measure for evaluating inspiratory effort in children. PARTICIPANTS: We studied 15 healthy children (age range, 5 to 12 years; mean age [+/- SD], 8.3 +/- 2.74; 9 male children) selected from patients referred to our pediatric center for routine assessment. MEASUREMENTS AND RESULTS: We assessed changes in the PTT during breathing against known resistances in awake children. Resistance was applied to the nose and mouth with a modified, two-way, nonrebreathing facemask. Our data show a good correlation between the induced inspiratory effort and the amplitude of PTT variations. CONCLUSIONS: PTT should be a useful method for quantifying changes in inspiratory effort due to augmented upper airway resistance in awake children. PMID- 14555585 TI - Influence of body composition, hemoglobin concentration, and cardiac size and function of gender differences in maximal oxygen uptake in prepubertal children. AB - OBJECTIVE: To determine the relative contribution of the gender difference in body composition, blood hemoglobin concentration, and cardiac dimension and function at rest and exercise of the gender difference in maximal oxygen uptake (O(2)max) in 10- to 12-year-old children. SUBJECTS: Thirty-five healthy children (17 girls and 18 boys; mean +/- SD age, 10.5 +/- 0.4 years). EXPERIMENTAL DESIGN: An anthropometric evaluation (body surface area, body fat content, and lean body mass [LBM]), assessment of hemoglobin concentration, echocardiographic evaluation at rest (left ventricular dimensions, and diastolic and systolic indexes at rest), and cardiovascular evaluation during a maximal cycle exercise (stroke volume [SV], total peripheral resistance). RESULTS: The boys exhibited a higher mass-relative O(2)max than the girls (47.9 mL/kg/min vs 40.9 mL/kg/min, respectively); but when normalized for LBM (allometric equation), the difference totally disappeared (19.0 mL/kg LBM(1.33)/min vs 18.9 mL/kg LBM(1.33)/min, respectively). No significant gender differences were seen in maximal heart rate and arteriovenous oxygen difference; however, maximal SV (SVmax) was significantly higher in boys than in girls, but when expressed relative to LBM, the difference was no longer significant. CONCLUSIONS: These findings demonstrate that contrary to adults, the sole limiting factor of O(2) that distinguished boys from girls was a lower SVmax in the latter; however, this gender difference totally disappeared when normalized for LBM. Consequently, the gender difference in heart size and cardiac function during exercise should be interpreted as only one aspect of the lower LBM in girls and not as reflective of a more basic functional gender difference. PMID- 14555586 TI - Reduction in ventilatory response to CO2 with relaxation feedback during CO2 rebreathing for ventilator patients. AB - BACKGROUND: Previous studies have shown that relaxation biofeedback reduced the amount of time spent receiving ventilation for difficult-to-wean patients. OBJECTIVE: The present study was begun to test the hypothesis that the underlying mechanism of biofeedback ventilator weaning was the reduction of neural respiratory drive (NRD). DESIGN: Prospective. SETTING: Pulmonary Medicine division in a Veterans Affairs hospital and the St. Louis Regional Medical Center. SUBJECTS: Twenty-four patients who were receiving mechanical ventilation were randomly assigned to either the biofeedback group or the control group. INTERVENTION: Respiratory relaxation feedback (RFB) was administered while a single variable, PaCO(2), was input to the respiratory control system and the output was measured. While rebreathing 7% CO(2)/93% O(2), the biofeedback group received a CO(2) trial session and a CO(2) RFB session, and the control group received a CO(2) trial session and a CO(2) no-feedback (NFB) session. MEASUREMENTS AND RESULTS: There was a significant (p < 0.05) reduction in respiratory and EEG parameters for the RFB group at maximal end-tidal CO(2) (mean [+/- SE], 70 +/- 0.2 mm Hg) between the CO(2) trial and the CO(2) RFB session compared to the control group where there was no significant difference between the results of the CO(2) trial and the CO(2) NFB session. The mean values for the CO(2) trial and CO(2) RFB session for the biofeedback group were as follows: occlusion pressure 0.1 s from the onset of inspiration, 8.42 +/- 1.08 and 6.48 +/ 0.78 cm H(2)O (which reflects NRD), respectively; minute inspiratory ventilation, 15.84 +/- 0.81 and 13.91 +/- 0.72 L/min, respectively; mean inspiratory flow, 670 +/- 2.28 and 581 +/- 35 mL/s, respectively; respiration rate, 32 +/- 2.28 and 31.2 +/- 2.58 breaths/min, respectively; and chest background electromyography, 4.89 +/- 0.71 and 3.54 +/- 0.54 microV, respectively. The mean electroencephalograph outputs for the CO(2) trial and CO(2) RFB session for the biofeedback group were as follows: mean EEG frequency, 14.78 +/- 0.98 and 13.06 +/- 0.59 Hz, respectively; and beta EEG power, 3.1 +/- 0.03 and 2.39 +/- 0.19, microV(2), respectively; and gamma EEG power, 2.96 +/- 0.34 and 2.24 +/- 0.24 microV(2), respectively. CONCLUSION: We conclude that the decrease in respiratory parameters reflecting NRD induced by RFB represents a key mechanism for the previously demonstrated effectiveness of biofeedback in reducing weaning time from mechanical ventilation. PMID- 14555587 TI - Clinical ventilator adjustments that improve speech. AB - STUDY OBJECTIVES: We sought to improve speech in tracheostomized individuals receiving positive-pressure ventilation. Such individuals often speak with short phrases, long pauses, and have problems with loudness and voice quality. SUBJECTS: We studied 15 adults with spinal cord injuries or neuromuscular diseases receiving long-term ventilation. INTERVENTIONS: The ventilator was adjusted using lengthened inspiratory time (TI), positive end-expiratory pressure (PEEP), and combinations thereof. RESULTS: When TI was lengthened (by 8 to 35% of the ventilator cycle), speaking time increased by 19% and pause time decreased by 12%. When PEEP was added (5 to 10 cm H(2)O), speaking time was 25% longer and obligatory pauses were 21% shorter. When lengthened TI and PEEP were combined (with or without reduced tidal volume), their effects were additive, increasing speaking time by 55% and decreasing pause time by 36%. The combined intervention improved speech timing, loudness, voice quality, and articulation. Individual differences in subject response to the interventions were substantial in some cases. We also tested high PEEP (15 cm H(2)O) in three subjects and found speech to be essentially identical to that produced with a one-way valve. CONCLUSIONS: These simple interventions markedly improve ventilator-supported speech and are safe, at least when used on a short-term basis. High PEEP is a safer alternative than a one-way valve. PMID- 14555588 TI - Development of a methacholine challenge method to minimize methacholine waste. AB - BACKGROUND: The standard 2-min tidal breathing methacholine challenge utilizes 3 mL to produce an output of 0.26 mL per 2 min, resulting in a substantial amount of methacholine being discarded. OBJECTIVE: To develop a method with reduced methacholine waste and to compare it to the standard method. METHODS: Twelve subjects with mild, well-controlled asthma volunteered for this investigation. They underwent three methacholine challenges in random order. The first challenge was the conventional 2-min tidal breathing method using 3 mL of doubling concentrations inhaled for 2 min at 5-min intervals. The first modification utilized 1.5 mL of quadrupling concentrations inhaled for 1 min and then 2 min, keeping the time interval constant at 3 min between completion of one inhalation and commencement of the next inhalation. The second modification utilized 1.5 mL of eightfold concentration step-ups inhaled for 30 s, 60 s, and 120 s with a time interval of 3 min between completion of one inhalation and commencement of the next inhalation. For each method, the provocative concentration of methacholine causing a 20% fall in FEV(1) (PC(20)) was calculated based on a 2-min equivalent dose inhalation. RESULTS: There was no significant difference in the geometric mean PC(20) (1.5 mg/mL, 1.6 mg/mL, and 1.6 mg/mL for the three methods, respectively; p = 0.47). The quadrupling concentration method was preferred because it was less subject to error than the other modification. CONCLUSION: The amount of methacholine discarded during a methacholine challenge can be reduced by two thirds by decreasing the volume from 3 to 1.5 mL, and by using quadrupling concentrations inhaled either with quadrupling-dose step-ups, or with doubling dose step-ups by using sequential 1-min and 2-min inhalations. PMID- 14555589 TI - Inhibition of cytokine release from alveolar macrophages in pulmonary sarcoidosis by pentoxifylline: comparison with dexamethasone. AB - STUDY OBJECTIVES: Pentoxifylline (POF) has been shown to suppress the cytokine production from lipopolysaccharide (LPS)-stimulated monocytes/alveolar macrophages (AMs). Sarcoidosis is a granulomatous disease that is driven by the action of tumor necrosis factor (TNF)-alpha and other proinflammatory cytokines. In this study, we aimed to investigate the effects of POF on the production of TNF-alpha, interleukin (IL)-1 beta, IL-6, IL-8, IL-10, and the soluble TNF receptors (sTNFRs) 1 and 2 from AMs in sarcoidosis, and we also compared them with those of dexamethasone (DEX). METHODS: AMs from 14 patients with sarcoidosis were cultured for 24 h with RPMI medium alone or with LPS (100 ng/mL), and with POF at concentrations of 0.01, 0.1, and 1 mmol/L, or with 0.1 mmol/L DEX. Cytokines in the culture supernatants were analyzed by enzyme-linked immunosorbent assay. RESULTS: The results showed that POF induced a dose dependent suppression of the spontaneous TNF-alpha release from AMs in sarcoidosis (p < 0.001), and that the spontaneous release of the other cytokines was unaffected by POF at all tested concentrations, but a trend for the inhibition of IL-10 production was found (p = 0.092). DEX inhibited the spontaneous release of TNF-alpha (p < 0.001), sTNFR2 (p < 0.05), IL-1 beta (p < 0.05), and IL-10 (p < 0.01). POF also suppressed the LPS-stimulated production of these cytokines except for that of sTNFR1. Similar to POF, DEX inhibited the LPS stimulated production of these cytokines, but not that of sTNFR1 and IL-1 beta. CONCLUSIONS: Compared with DEX, POF may improve therapeutic regimens in patients with sarcoidosis either by sparing or by replacing corticosteroids. However, the precise clinical value of POF in the treatment of sarcoidosis and other lung diseases will have to be determined in further clinical trials. PMID- 14555590 TI - Sore throat, dysphagia, stiffness in back of neck, and dyspnea following tricyclic antidepressant overdose. PMID- 14555591 TI - Measuring agreement between diagnostic devices. AB - There is growing interest in using portable monitoring for investigating patients with suspected sleep apnea. Research studies typically report portable monitoring results in comparison with the results of sleep laboratory-based polysomnography. A systematic review of this research has recently been completed by a joint working group of the American College of Chest Physicians, the American Thoracic Society, and the American Academy of Sleep Medicine. The methods for comparing the results of portable monitors and polysomnography include product-moment correlation, intraclass correlation, mean differences/limits of agreement, sensitivity, specificity, and likelihood ratios. Each approach has advantages and limitations, which are highlighted in this review. PMID- 14555592 TI - Home diagnosis of sleep apnea: a systematic review of the literature. An evidence review cosponsored by the American Academy of Sleep Medicine, the American College of Chest Physicians, and the American Thoracic Society. PMID- 14555594 TI - Inhaled fluticasone propionate by diskus in the treatment of asthma: a comparison of the efficacy of the same nominal dose given either once or twice a day. AB - STUDY OBJECTIVE: In September 2000, the US Food and Drug Administration (FDA) approved the use of Flovent Diskus (FD) [fluticasone propionate; GlaxoSmithKline; Research Triangle Park, NC], which is an orally inhaled, dry-powder corticosteroid, for the maintenance treatment of asthma at dosages of 50 to 1,000 microg administered twice-daily. Once-daily dosage regimens did not receive approval. This article will detail six clinical trials, five of which incorporated comparative once-daily and twice-daily treatment arms of the same nominal dose of FD. DESIGN: Six 12-week, randomized, double-blind, placebo controlled studies in patients with mild-to-moderate asthma, including two pediatric asthma trials (patient age, 4 to 11 years) of total daily doses of fluticasone propionate (FP) of 100 or 200 microg, and four adult and adolescent studies of total daily doses of FP of 100, 200, or 500 microg. RESULTS: Twice daily dosing was numerically superior to once-daily dosing at the same nominal dose in all comparative studies for the primary end point, change in predose FEV(1). In five trials, the results of the once-daily dosage of FP were statistically indistinguishable from those with placebo. One trial demonstrated the superiority of FP, 500 microg once-daily, over placebo; however, the effect size was half that observed with twice-daily dosing. Once-daily FP dosing showed no advantage in safety or in patient adherence to medication. CONCLUSIONS: In the FDA review of once-daily dosing of the FD regimen, 100 or 200 microg once-daily dosing was not shown to be significantly better than placebo. FP 500 microg once daily was found to be superior to placebo, but at about one half the effect size as the same nominal dose given bid. No advantage in patient safety or adherence was demonstrated for once-daily administration over twice-daily administration, and once-daily administration is not currently recommended. PMID- 14555595 TI - An unusual case of travel sickness. PMID- 14555596 TI - Fever, consciousness disturbance, and muscle rigidity in a 68-year-old man with depressive disorder. PMID- 14555597 TI - Extrathoracic obstruction and hypoxemia occurring during exercise in a competitive female cyclist. AB - A 22-year-old competitive female cyclist complained of cough, chest tightness, and wheeze during high-intensity exercise that had previously been diagnosed as exercise-induced bronchospasm (EIB). A loud stridor, a sensation of her "throat closing," and severe dyspnea developed during maximal cycling exercise with concomitant reductions in both inspiratory and expiratory flow rates. A decrease of 25 L/min (26%) in minute ventilation and arterial hypoxemia (PaO(2) decrease, 93 to 76.5 mm Hg) resulted from this obstruction. Spontaneous tidal flow-volume loops (FVLs) during exercise exhibited a sawtooth pattern during inspiration, and substantial drops in flow rates after the stridor developed. However, maximal FVLs were unchanged from baseline following exercise, indicating that the obstruction was not EIB. We suggest that the continuous measurement of spontaneous breath-by-breath tidal FVLs may be a useful diagnostic tool for the identification of exercise-induced extrathoracic obstruction. Additionally, extrathoracic obstruction should be considered as an uncommon but potential cause of inadequate ventilation and arterial hypoxemia during exercise. PMID- 14555598 TI - High dose rate brachytherapy for nonmalignant airway obstruction: new treatment option. AB - STUDY OBJECTIVES: High dose rate (HDR) endobronchial brachytherapy is widely used as a palliative treatment for symptomatic airway obstruction by primary or secondary malignant tumors. We report on a successful use of HDR brachytherapy in patients with nonmalignant airway obstruction. DESIGN: Case series PATIENTS: Six patients received HDR brachytherapy for airway obstruction caused by granulation tissue around a metal stent placed for restoration of the airway patency for nonmalignant causes. In four patients, brachytherapy was performed following recurrent occlusion of the airway by granulation tissue formation; in two patients, it was done as a prophylactic procedure. INTERVENTION: HDR brachytherapy catheters were passed through the metal stents under direct fluoroscopic guidance. Simulation and computerized treatment planning were done, and a single dose of 10 Gy was administered using a brachytherapy remote afterloader with a (192)Ir source. The dose was prescribed to a distance of 1 cm from the center of the source, with a margin of 1 cm from the proximal and distal ends of the stent. RESULTS: At a median follow-up of 15 months, moderate granulation tissue formation was observed in only one patient; in four others, it was categorized as minimal, 5 to 30 months from the procedure. Restoration of the lumen was complete in four patients, near complete in one patient, and partial in one patient. In one patient, previously treated by external radiotherapy, local tissue necrosis was evident. CONCLUSION: HDR brachytherapy can be used safely for nonmalignant airway obstruction. Further studies including more patients and longer follow-up are needed. PMID- 14555599 TI - A mycotic pulmonary artery aneurysm presenting as an endobronchial mass. AB - We report the case of a 49-year-old woman who developed an endobronchial mycotic pulmonary artery aneurysm (PAA) as a complication of pneumonia. After presenting with patchy infiltrates, she developed right lung atelectasis. A noncontrast chest CT scan revealed a mass in the right hilum, and bronchoscopy identified an obstructing lesion in the right mainstem bronchus. Unfortunately, biopsy caused massive hemorrhage, and the patient died. An autopsy identified a mycotic PAA that had invaded the right mainstem. The case suggests that a contrasted CT scan should be included in the evaluation of endobronchial masses and that PAA should be included in the differential diagnosis of such lesions. PMID- 14555600 TI - Successful withdrawal of long-term epoprostenol therapy for pulmonary arterial hypertension. AB - BACKGROUND: IV epoprostenol treatment of pulmonary arterial hypertension (PAH) has been believed to require an indefinite duration of therapy OBJECTIVE: To describe the successful discontinuation of long-term epoprostenol therapy in four patients DESIGN: Case reports SETTING: Outpatient clinic, tertiary-care hospital PATIENTS: Four patients with acutely nonvasoreactive, World Health Organization (WHO) functional class IV PAH received long-term epoprostenol therapy. All patients subsequently demonstrated normalization of pulmonary arterial pressures on epoprostenol treatment. These patients were selected for epoprostenol withdrawal INTERVENTION: Down-titration and discontinuation of epoprostenol RESULTS: All four patients were safely transitioned from epoprostenol to oral therapies and have maintained WHO functional class I-II for a mean of 11 months (range, 8 to 16 months). The duration of epoprostenol therapy prior to discontinuation averaged 5.7 years (range, 2.4 to 13.5 years) CONCLUSION: Epoprostenol may sufficiently reverse the pathogenic process in select patients with PAH to allow a transition to less complex and less invasive treatment modalities. PMID- 14555601 TI - The long-term results of gastric bypass on indexes of sleep apnea. AB - STUDY OBJECTIVES: Weight loss improves obstructive sleep apnea (OSA), and clinicians regularly recommend dieting to their patients with OSA; however, many morbidly obese patients may be unable to lose weight without medical or surgical intervention. Gastric bypass surgery (GB) facilitates weight loss for morbidly obese patients. Studies show GB will improve symptoms associated with OSA, but little is known regarding the long-term effects of GB on this population. DESIGN: Historical, retrospective, cohort study. SETTING: Sleep Disorders Center, Sentara Norfolk General Hospital, Norfolk, VA. PARTICIPANTS: A list of subjects who underwent vertical Roux-en-Y GB was cross-referenced with the Sleep Disorders Center at Sentara Norfolk General Hospital to identify patients with a diagnosis of OSA during the preoperative evaluation prior to undergoing GB. INTERVENTIONS: GB. MEASUREMENTS: Our primary end point was the respiratory disturbance index (RDI). Secondary variables were body mass index (BMI), mean oxygen saturation, low oxygen saturation, a standardized depression scale, and the continuous positive airway pressure (CPAP) requirement. RESULTS: Thirty-four subjects with a diagnosis of OSA prior to GB were identified. Of these, 28 subjects were located and offered follow-up polysomnography after GB. Eight subjects returned for repeat polysomnography. The subjects were re-evaluated an average of 28 months after GB. Seven subjects had a lower BMI after GB. Mean BMI was reduced by 31% (p = 0.001). The mean decrease in RDI was 75% (p = 0.01), and five of the eight subjects no longer required nasal CPAP. Mean nocturnal oxygen saturation improved from 95 to 97% (p = 0.04). CONCLUSIONS: Weight reduction following GB is associated with significant improvements in sleep apnea indexes an average of 28 months after GB. Re-evaluation after GB is necessary to identify and treat those patients who, despite subjective improvement, may continue to require CPAP for residual OSA. PMID- 14555602 TI - High-altitude pulmonary edema. PMID- 14555603 TI - Interleukin-6, obstructive sleep apnea, and obesity. PMID- 14555604 TI - Beta-blockers with vasodilatory actions. PMID- 14555605 TI - Regards to Bill. PMID- 14555606 TI - Evaluation of diagnostic device for obstructive sleep apnea. PMID- 14555609 TI - Evaluation of the cancer chemopreventive potency of dithiolethione analogs of oltipraz. AB - Oltipraz and related dithiolethiones constitute an important class of chemopreventive agents that enhance the expression of carcinogen detoxication and antioxidant genes. Dose-response studies were undertaken to characterize the cancer chemopreventive activities of several dithiolethiones that are at least as active as oltipraz as inducers. Inhibition of formation of pre-neoplastic lesions and formation of DNA adducts in livers of rats exposed to aflatoxin B1 (AFB1) was monitored. In the tumorigenesis experiment, the dithiolethiones were orally gavaged 3 days/week for 3 successive weeks and at four doses ranging from 0.03 to 0.3 mmol/kg body wt. AFB1 was gavaged beginning 1 week after the start of the dithiolethiones and for two successive weeks. The burden of AFB1-induced putative pre-neoplastic lesions (glutathione S-transferase-placental isoform positive foci) was quantified by light microscopy. Reduction in AFB-DNA adduct burden was assessed 24 h following the first dose of AFB1. Both the parent 1,2-dithiole-3 thione (D3T) and its 5-tert-butyl derivative were more potent inhibitors than oltipraz against these endpoints, while two of the seven tested analogs were slightly less inhibitory. D3T, the most potent dithiolethione of this series, was examined by microarray analysis for induction of hepatic genes at an intermediate chemopreventive dose (0.1 mmol/kg). Transcript levels of eight genes, including two known to detoxify aflatoxin, namely, glutathione S-transferase A5 (GSTA5) and AFB1 aldehyde reductase (AFAR) were elevated. Western analysis indicated that induction of hepatic GSTA5 and AFAR were directly related to the dose of D3T. At the highest dose of D3T (0.3 mmol/kg), protein levels of GSTA5 and AFAR were induced by 7- and 27-fold, respectively. While efficacy in humans has yet to be tested, D3T is clearly more potent than oltipraz and serves as a useful molecular probe for determining the key events associated with protection by this class of agents. PMID- 14555610 TI - Diallyl disulfide (DADS) enhances gap-junctional intercellular communication by both direct and indirect mechanisms in rat liver cells. AB - Diallyl disulfide (DADS), a sulfur compound from garlic has been shown to exert many biological effects: induction of carcinogen detoxication, inhibition of tumor cell proliferation, etc. These effects are consistent with its anticarcinogenic properties in animal models and could account for garlic protective effects in humans. Our study demonstrates that DADS can improve gap junctional intercellular communication (GJIC) in vitro. In rat liver epithelial cells (REL cells), using the dye transfer assay, we observe a time-dependent stimulation of GJIC by DADS at non-cytotoxic concentrations. In addition, incubation of cells with DADS for 1 h prevents the inhibition of GJIC induced by 3,5-di-tertio-butyl-4-hydroxytoluene (BHT). We have studied the direct effects of DADS on the regulation of GJIC, and especially on the expression and localization of the connexin expressed in these cells (Cx43): the enhancement of dye transfer (x1.6) by DADS from 1 to 50 micro M is associated with an increase (x1.3-1.8) in the amount of Cx43 protein (western blotting) with no alteration of its localization in the cell-cell contact regions of the plasma membrane (immunofluorescence analysis). We have also explored the possibility that DADS might act indirectly on GJIC. On one hand, DADS does not change the amount of E cadherin, the adhesion molecule expressed in epithelial cells. On the other hand, it induces rapid inhibition of protein glycosylation. The data suggest that DADS could reduce local constraints imposed by glycoproteins, thus facilitating dye transfer. In conclusion, DADS can be included with other plant microconstituents, which have been demonstrated to improve GJIC. Its effect on REL cells can be explained by its ability to enhance the amount of Cx43 and also to diminish the level of glycosylated proteins. PMID- 14555611 TI - Characterizing the role of MDM2 in diethylnitrosamine induced acute liver damage and development of pre-neoplastic lesions. AB - Pre-neoplastic lesions in rodent liver often express high levels of MDM2 and lack a p53 response to DNA damage. The question we posed was whether there is a liver specific regulation of the p53/MDM2 feedback loop and if it can be related to the development of pre-neoplastic lesions, referred to as enzyme altered foci (EAF) in rats. Acute responses of p53 and MDM2 to diethylnitrosamine (DEN) were characterized by employing immunohistochemistry, western blotting, RT-PCR and in situ hybridization. A single dose of DEN induced a centrilobular p53 response that peaked at 24 h. It was associated with transcriptional activation of MDM2 and signs of apoptosis. However, in midzonal hepatocytes, which constitutively expressed high levels of cytoplasmic MDM2, there was a rapid-onset but transient p53 response. It was terminated at 24 h and there were no signs of apoptosis. The rapidly declining p53 levels in midzonal areas was preceded by a transient peak in MDM2 mRNA levels at 6 h. Rats pre-treated with repeated low or high weekly doses of DEN exhibited EAF and these lesions expressed high levels of cytoplasmic MDM2. Using MDM2 as a marker for EAF gave similar results as using glutathione transferase-P (GST-P) as a marker. Furthermore, small EAF, elicited by low doses of DEN, were preferentially localized to midzonal areas. It is concluded that in centrilobular areas DEN-induced alterations in p53/MDM2 levels are compatible with a previously described feedback loop. An attenuated p53 response in midzonal hepatocytes can be related to a high constitutive expression of MDM2 in these cells. The localization of small EAF to midzonal areas, and the fact that EAF cells expressed high levels of MDM2, indicates that MDM2 expression is a factor governing initiation and early development of EAF. The data support the hypothesis that EAF hepatocytes are initiated via epigenetic mechanisms. PMID- 14555612 TI - The role of p53 in base excision repair following genotoxic stress. AB - The p53 tumor suppressor protein is involved in apoptosis and cell cycle checkpoints. We have shown recently that p53 also facilitates base excision repair (BER). To further examine p53 involvement in the regulation of BER we chose to focus on 3-methyladenine DNA glycosylase (3-MeAde DNA glycosylase), the first enzyme acting in the BER pathway. 3-MeAde DNA glycosylase activity was found to be modulated by the p53 protein. This modulation was dependent on the type of genotoxic stress used. Gamma-irradiation damage resulted in activation of glycosylase, which was enhanced by p53. Doxorubicin and hydrogen peroxide (H2O2) treatment, although inducing p53 stabilization, did not cause the activation of glycosylase. Nitric oxide (NO) resulted in activation of 3-MeAde DNA glycosylase. Surprisingly this activation was down regulated by wild-type p53. The down regulation of 3-MeAde DNA glycosylase activity was due to trans repression of glycosylase mRNA by p53. Furthermore, we found that AP endonuclease (APE) activity was not altered by NO. Our study provides evidence for a possible antimutagenic role for p53 following exposure of cells to NO species. In the absence of p53, NO exposure results in elevation of 3-MeAde DNA glycosylase activity that results in elevation in the number of AP sites in DNA. At the same time, APE activity does not rise and removal of the AP sites is not further processed resulting in a mutator phenotype. When p53 is present, it down regulates the transcription of 3-MeAde DNA glycosylase. This provides a new model by which p53 prevents the creation of a mutator phenotype. PMID- 14555613 TI - Alteration of gene expression during radiation-induced resistance and tumorigenesis in NIH3T3 cells revealed by cDNA microarrays: involvement of MDM2 and CDC25B. AB - To identify a set of genes involved in the development of radiation-induced tumorigenesis, we used DNA microarrays consisting of 1176 mouse genes and compared expression profiles of radioresistant cells, designated NIH3T3-R1 and NIH3T3-R4. These cells were tumorigenic in a nude mouse grafting system, as compared with the parental NIH3T3 cells. Expression of MDM2, CDK6 and CDC25B was found to increase more than 3-fold. Entactin protein levels were down-regulated in NIH3T3-R1 and NIH3T3-R4 cells. Changes in gene expression were confirmed by reverse transcription-PCR or western blotting. When these genes were transfected into NIH3T3 cells, CDC25B and MDM2 overexpressing NIH3T3 cells showed radioresistance, while CDK6 overexpressing cells did not. In the case of entactin, overexpressing NIH3T3-R1 and NIH3T3-R4 cells were still radioresistant. Furthermore, CDC25B and MDM2 overexpressing cells grafted into nude mice were tumorigenic. NIH3T3-R1 and NIH3T3-R4 cells showed increased radiation-induced apoptosis accompanied by a faster growth rate, rather than an earlier radiation induced G2/M phase arrest, suggesting that the radioresistance of NIH3T3-R1 and NIH3T3-R4 cells was due to a faster growth rate rather than induction of apoptosis. In the case of MDM2 and CDC25B overexpressing cells, similar phenomena, such as increased apoptosis and a faster growth rate, were shown. The above results, therefore, demonstrate involvement of CDC25B and MDM2 overexpression in radiation-induced tumorigenesis and provide novel targets for detection of radiation-induced carcinogenesis. PMID- 14555614 TI - Dual efficacy of silibinin in protecting or enhancing ultraviolet B radiation caused apoptosis in HaCaT human immortalized keratinocytes. AB - An increasing incidence of human skin cancer and other adverse effects of solar ultraviolet (UV) radiation enhance the need for novel chemoprevention strategies. Here, we have studied the effect of silibinin on UVB-induced apoptosis in HaCaT cells. Silibinin strongly prevented lower doses (15 and 30 mJ/cm2) of UVB-induced apoptosis, as observed by a reversal in UVB-caused poly(ADP-ribose) polymerase (PARP) cleavage, caspase 9 activation and an increase in apoptotic cells. UVB induced PARP cleavage was also abolished by all caspase inhibitor, suggesting that it is a caspase-dependent effect. In other studies, silibinin restored UVB caused depletion of a protein inhibitor of apoptosis, survivin, concomitant with up-regulation of transcription factor nuclear factor kappaB DNA binding activity, without any noticeable effect on UVB-caused activated protein-1 activation. Further, silibinin treatment up-regulated UVB-induced extracellular signal regulated kinase 1/2 phosphorylation, suggesting a possible role as a survival event in the protective effect of silibinin. In other studies, silibinin caused a moderate increase in phospho-Bcl-2, without any noticeable changes in total Bcl-2 levels, and down-regulated bax levels moderately. Silibinin also caused a strong decrease in Bad heterodimerization with Bclx(L), which was consistent with an increased translocation of Bclx(L) to the mitochondria from the cytosol. Consistent with its protective effect on UVB-caused apoptosis, silibinin also increased S phase arrest, possibly providing a prolonged time for efficient DNA repair. Interestingly, the protective effects of silibinin in HaCaT cells were lost at a higher dose of UVB (120 mJ/cm2) and instead it further enhanced UVB caused apoptosis together with a strong decrease in UVB-caused activated protein 1 activation. Together, these results clearly demonstrate the dual efficacy of silibinin in protecting or enhancing UVB-caused apoptosis in the same cellular system and suggest that silibinin possibly works as a UVB damage sensor to exert its biological action. PMID- 14555615 TI - Plasma isoflavone levels versus self-reported soy isoflavone levels in Asian American women in Los Angeles County. AB - In a case-control study conducted among Asian-American women in Los Angeles County, we reported that the risk of breast cancer was significantly reduced in association with soy intake [Wu,A.H., Wan,P., Hankin,J. et al. (2002) Carcinogenesis, 23, 1491-1496]. In a subset of cases (n = 97) and controls (n = 97) we investigated the relationship between self-reported usual adult intake of soy isoflavones which was determined from a food frequency questionnaire and levels of plasma isoflavones (genistein and daidzein) and isoflavone metabolites (equol, dihydrogenistein and dihydrodaidzein) from a randomly timed blood specimen. In analyses conducted in cases and controls separately, levels of plasma genistein, daidzein and total isoflavones increased with increasing levels of self-reported intake of soy isoflavones. Breast cancer cases and control subjects did not differ in their respective associations between total plasma isoflavone levels and self-reported intake (P = 0.48). Among all subjects, there was a 3-fold difference in geometric mean plasma levels of total isoflavones [81.8 (95% CI = 53.4, 125.1) versus 26.4 nmol/l (95% CI = 16.6, 41.8)] between women in the highest quartile of soy isoflavone intake (>12.68 mg isoflavones/1000 kcal) compared with those in the lowest quartile of intake ( Asp mutation in PBP 5 markedly impairs deacylation with only minor effects on acylation, and abolishes CPase activity. We have determined the three-dimensional structure of a soluble form of wild-type PBP 5 at 1.85-A resolution and have also refined the structure of the G105D mutant form of PBP 5 to 1.9-A resolution. Comparison of the two structures reveals that the major effect of the mutation is to disorder a loop comprising residues 74-90 that sits atop the SXN motif of the active site. Deletion of the 74-90 loop in wild-type PBP 5 markedly diminished the deacylation rate of penicillin G with a minimal impact on acylation, and abolished CPase activity. These effects were very similar to those observed in the G105D mutant, reinforcing the idea that this mutation causes disordering of the 74-90 loop. Mutation of two consecutive serines within this loop, which hydrogen bond to Ser110 and Asn112 in the SXN motif, had marked effects on CPase activity, but not beta-lactam antibiotic binding or hydrolysis. These data suggest a direct role for the SXN motif in deacylation of the acyl-enzyme complex and imply that the functioning of this motif is modulated by the 74-90 loop. PMID- 14555649 TI - Delta Np63 alpha expression is regulated by the phosphoinositide 3-kinase pathway. AB - p63 is a homologue of p53 that functions to maintain progenitor cell populations in stratified epithelia. Delta Np63 alpha is overexpressed in epithelial cancers and has been shown to have oncogenic properties. We have previously reported that inhibition of epidermal growth factor receptor signaling results in a decrease in Delta Np63 alpha expression. Here, we demonstrate Delta Np63 alpha is a target of the phosphoinositide-3-kinase (PI3K) pathway downstream of the epidermal growth factor receptor. Treatment of keratinocytes with epidermal growth factor results in an increase in Delta Np63 alpha expression at the mRNA level, which is abrogated by inhibition of PI3K but not mitogen-activated protein kinase signaling. Small interfering RNA-mediated knockdown of the p110 beta catalytic subunit of PI3K results in a decrease in Delta Np63 alpha protein levels in keratinocytes. The results presented herein suggest that regulation of Delta Np63 alpha expression by the PI3K pathway plays a critical role in the survival and proliferative capacity of squamous epithelia. PMID- 14555650 TI - Atomic resolution structure of the HFBII hydrophobin, a self-assembling amphiphile. AB - Hydrophobins are proteins specific to filamentous fungi. Hydrophobins have several important roles in fungal physiology, for example, adhesion, formation of protective surface coatings, and the reduction of the surface tension of water, which allows growth of aerial structures. Hydrophobins show remarkable biophysical properties, for example, they are the most powerful surface-active proteins known. To this point the molecular basis of the function of this group of proteins has been largely unknown. We have now determined the crystal structure of the hydrophobin HFBII from Trichoderma reesei at 1.0 A resolution. HFBII has a novel, compact single domain structure containing one alpha-helix and four antiparallel beta-strands that completely envelop two disulfide bridges. The protein surface is mainly hydrophilic, but two beta-hairpin loops contain several conserved aliphatic side chains that form a flat hydrophobic patch that makes the molecule amphiphilic. The amphiphilicity of the HFBII molecule is expected to be a source for surface activity, and we suggest that the behavior of this surfactant is greatly enhanced by the self-assembly that is favored by the combination of size and rigidity. This mechanism of function is supported by atomic force micrographs that show highly ordered arrays of HFBII at the air water interface. The data presented show that much of the current views on structure function relations in hydrophobins must be re-evaluated. PMID- 14555651 TI - EVI1 promotes cell proliferation by interacting with BRG1 and blocking the repression of BRG1 on E2F1 activity. AB - EVI1 is a complex protein required for embryogenesis and inappropriately expressed in many types of human myeloid leukemia. Earlier we showed that the forced expression of EVI1 in murine hematopoietic precursor cells leads to their abnormal differentiation and increased proliferation. In this report, we show that EVI1 physically interacts with BRG1 and its functional homolog BRM in mammalian cells. We found that the C terminus of EVI1 interacts strongly with BRG1 and that the central and C-terminal regions of BRG1 are involved in EVI1 BRG1 interaction. Using reporter gene assays, we demonstrate that EVI1 activates the E2F1 promoter in NIH3T3 cells but not in BRG1-negative SW13 cells. Ectopic expression of BRG1 is able to repress the E2F1 promoter in vector-transfected SW13 cells but not in EVI1-transfected SW13 cells. Finally, we show that EVI1 up regulates cell proliferation in BRG1-positive 32Dcl3 cells but not in BRG1 negative SW13 cells. Taken together, these data support the hypothesis that the interaction with BRG1 is important for up-regulation of cell-growth by EVI1. PMID- 14555652 TI - Estradiol binding to maxi-K channels induces their down-regulation via proteasomal degradation. AB - Estrogens exert their biological action via both genomic and non-genomic mechanisms. Proteins different from classical estradiol receptors are believed to mediate the latter effects. Here we demonstrate that the maxi-K channel functions as an estrogen-binding protein in transfected HEK293 cells. Whole-cell maxi-K channel currents and protein expression were attenuated by exposure to either 17alpha- or 17beta-estradiol. This effect was dose-dependent for 17beta-estradiol at concentrations ranging from 10 nm to 1 microm, while 17alpha-estradiol inhibited channel expression only at 1 microm. These effects were mediated by direct low affinity binding of estradiol to the maxi-K channel but not to its accessory beta1-subunit, as revealed by cell membrane estradiol binding assays. However, specific binding of estradiol to the channel was facilitated by the presence of the beta1 subunit. Addition of MG-132, a blocker of proteasomal degradation, stabilized channel expression. These data suggest that channel down regulation is mediated by estrogen-induced proteasomal degradation, similar to the pathway used for estrogen receptor degradation. Membrane expression of endogenous maxi-K channels in cultured vascular smooth muscle cells was also attenuated by prolonged exposure to 17alpha- and 17beta-estradiol. Thus our studies demonstrate that estrogen binds to maxi-K channels and may directly regulate channel expression and function. These results will have important implications in understanding estradiol-induced effects in multiple tissues including vascular smooth muscle. PMID- 14555653 TI - LEK1 is a potential inhibitor of pocket protein-mediated cellular processes. AB - LEK1, a member of the LEK family of proteins, is ubiquitously expressed in developing murine tissues. Our current studies are aimed at identifying the role of LEK1 during cell growth and differentiation. Little is known about the function of LEK proteins. Recent studies in our laboratory have focused on the characterization of the LEK1 atypical Rb-binding domain that is conserved among all LEK proteins. Our findings suggest that LEK1 potentially functions as a universal regulator of pocket protein activity. Pocket proteins exhibit distinct expression patterns during development and function to regulate cell cycle, apoptosis, and tissue-specific gene expression. We show that LEK1 interacts with all three pocket proteins, p107, p130, and pRb. Additionally, this interaction occurs specifically between the LEK1 Rb-binding motif and the "pocket domain" of Rb proteins responsible for Rb association with other targets. Analyses of the effects of disruption of LEK1 protein expression by morpholino oligomers demonstrate that LEK1 depletion decreases cell proliferation, disrupts cell cycle progression, and induces apoptosis. Given its expression in developing cells, its association with pocket proteins, and its effects on proliferation, cell cycle, and viability of cells, we suggest that LEK1 functions in a similar manner to phosphorylation to disrupt association of Rb proteins with appropriate binding targets. Thus, the LEK1/Rb interaction serves to retain cells in a pre differentiative, actively proliferative state despite the presence of Rb proteins during development. Our data suggest that LEK1 is unique among LEK family members in that it specifically functions during murine development to regulate the activity of Rb proteins during cell division and proliferation. Furthermore, we discuss the distinct possibility that a yet unidentified splice variant of the closely related human CENP-F, serves a similar function to LEK1 in humans. PMID- 14555655 TI - Thermodynamic and structural analysis of peptide- and allele-dependent properties of two HLA-B27 subtypes exhibiting differential disease association. AB - Selected HLA-B27 subtypes are associated with spondyloarthropathies, but the underlying mechanism is not understood. To explain this association in molecular terms, a comparison of peptide-dependent dynamic and structural properties of the differentially disease-associated subtypes HLA-B*2705 and HLA-B*2709 was carried out. These molecules differ only by a single amino acid at the floor of the peptide binding groove. The thermostabilities of a series of HLA-B27 molecules complexed with nonameric and decameric peptides were determined and revealed substantial differences depending on the subtype as well as the residues at the termini of the peptides. In addition we present the crystal structure of the B*2709 subtype complexed with a decameric peptide. This structure provides an explanation for the preference of HLA-B27 for a peptide with an N-terminal arginine as secondary anchor and the lack of preference for tyrosine as peptide C terminus in B*2709. The data show that differences in thermodynamic properties between peptide-complexed HLA-B27 subtypes are correlated with a variety of structural properties. PMID- 14555654 TI - Riboflavin uptake and FAD synthesis in Saccharomyces cerevisiae mitochondria: involvement of the Flx1p carrier in FAD export. AB - We have studied the functional steps by which Saccharomyces cerevisiae mitochondria can synthesize FAD from cytosolic riboflavin (Rf). Riboflavin uptake into mitochondria took place via a mechanism that is consistent with the existence of (at least two) carrier systems. FAD was synthesized inside mitochondria by a mitochondrial FAD synthetase (EC 2.7.7.2), and it was exported into the cytosol via an export system that was inhibited by lumiflavin, and which was different from the riboflavin uptake system. To understand the role of the putative mitochondrial FAD carrier, Flx1p, in this pathway, an flx1Delta mutant strain was constructed. Coupled mitochondria isolated from flx1Delta mutant cells were compared with wild-type mitochondria with respect to the capability to take up Rf, to synthesize FAD from it, and to export FAD into the extramitochondrial phase. Mitochondria isolated from flx1Delta mutant cells specifically lost the ability to export FAD, but did not lose the ability to take up Rf, FAD, or FMN and to synthesize FAD from Rf. Hence, Flx1p is proposed to be the mitochondrial FAD export carrier. Moreover, deletion of the FLX1 gene resulted in a specific reduction of the activities of mitochondrial lipoamide dehydrogenase and succinate dehydrogenase, which are FAD-binding enzymes. For the flavoprotein subunit of succinate dehydrogenase we could demonstrate that this was not due to a changed level of mitochondrial FAD or to a change in the degree of flavinylation of the protein. Instead, the amount of the flavoprotein subunit of succinate dehydrogenase was strongly reduced, indicating an additional regulatory role for Flx1p in protein synthesis or degradation. PMID- 14555657 TI - In vitro selection of second site revertants analysis of the hairpin ribozyme active site. AB - We have used in vitro genetics to evaluate the function and interactions of the conserved base G8 in the hairpin ribozyme catalytic RNA. Second site revertant selection for a G8X mutant, where X is any of the other three natural nucleobases, yielded a family of second site suppressors of the G8U mutant, but not of G8C or G8A, indicating that only G and U can be tolerated at position 8 of the ribozyme. This result is consistent with recent observations that point to the functional importance of G8 N-1 in the chemistry of catalysis by this ribozyme reaction. Suppression of the G8U mutation was observed when changes were made directly across loop A from the mutated base at substrate position +2 or positions +2 and +3 in combination. The same changes made in the context of the natural G8 sequence resulted in a very large drop in activity. Thus, the G8U mutation results in a change in specificity of the ribozyme from 5'-N / GUC-3' to 5'-N / GCU-3'. The results presented imply that G8 interacts directly with U+2 during catalysis. We propose that this interaction favors the correct positioning of the catalytic determinants of G8. The implications for the folding of the ribozyme and the catalytic mechanism are discussed. PMID- 14555656 TI - Identification of Drosophila neuropeptide receptors by G protein-coupled receptors-beta-arrestin2 interactions. AB - Activation of G protein-coupled receptors (GPCR) leads to the recruitment of beta arrestins. By tagging the beta-arrestin molecule with a green fluorescent protein, we can visualize the activation of GPCRs in living cells. We have used this approach to de-orphan and study 11 GPCRs for neuropeptide receptors in Drosophila melanogaster. Here we verify the identities of ligands for several recently de-orphaned receptors, including the receptors for the Drosophila neuropeptides proctolin (CG6986), neuropeptide F (CG1147), corazonin (CG10698), dFMRF-amide (CG2114), and allatostatin C (CG7285 and CG13702). We also de-orphan CG6515 and CG7887 by showing these two suspected tachykinin receptor family members respond specifically to a Drosophila tachykinin neuropeptide. Additionally, the translocation assay was used to de-orphan three Drosophila receptors. We show that CG14484, encoding a receptor related to vertebrate bombesin receptors, responds specifically to allatostatin B. Furthermore, the pair of paralogous receptors CG8985 and CG13803 responds specifically to the FMRF amide-related peptide dromyosuppressin. To corroborate the findings on orphan receptors obtained by the translocation assay, we show that dromyosuppressin also stimulated GTPgammaS binding and inhibited cAMP by CG8985 and CG13803. Together these observations demonstrate the beta-arrestin-green fluorescent protein translocation assay is an important tool in the repertoire of strategies for ligand identification of novel G protein-coupled receptors. PMID- 14555658 TI - Evaluation by mutagenesis of the importance of 3 arginines in alpha, beta, and gamma subunits of human NAD-dependent isocitrate dehydrogenase. AB - Mammalian NAD-dependent isocitrate dehydrogenase is an allosteric enzyme, activated by ADP and composed of 3 distinct subunits in the ratio 2alpha:1beta:1gamma. Based on the crystal structure of NADP-dependent isocitrate dehydrogenases from Escherichia coli, Bacillus subtilis, and pig heart, and a comparison of their amino acid sequences, alpha-Arg88, beta-Arg99, and gamma Arg97 of human NAD-dependent isocitrate dehydrogenase were chosen as candidates for mutagenesis to test their roles in catalytic activity and ADP activation. A plasmid harboring cDNA that encodes alpha, beta, and gamma subunits of the human isocitrate dehydrogenase (Kim, Y. O., Koh, H. J., Kim, S. H., Jo, S. H., Huh, J. W., Jeong, K. S., Lee, I. J., Song, B. J., and Huh, T. L. (1999) J. Biol. Chem. 274, 36866-36875) was used to express the enzyme in isocitrate dehydrogenase deficient E. coli. Wild type (WT) and mutant enzymes (each containing 2 normal subunits plus a mutant subunit with alpha-R88Q, beta-R99Q, or gamma-R97Q) were purified to homogeneity yielding enzymes with 2alpha:1beta:1gamma subunit composition and a native molecular mass of 315 kDa. Specific activities of 22, 14, and 2 micromol of NADH/min/mg were measured, respectively, for WT, beta-R99Q, and gamma-R97Q enzymes. In contrast, mutant enzymes with normal beta and gamma subunits and alpha-R88Q mutant subunit has no detectable activity, demonstrating that, although beta-Arg99 and gamma-Arg97 contribute to activity, alpha-Arg88 is essential for catalysis. For WT enzyme, the Km for isocitrate is 2.2 mm, decreasing to 0.3 mm with added ADP. In contrast, for beta-R99Q and gamma-R97Q enzymes, the Km for isocitrate is the same in the absence or presence of ADP, although all the enzymes bind ADP. These results suggest that beta-Arg99 and gamma-Arg97 are needed for normal ADP activation. In addition, the gamma-R97Q enzyme has a Km for NAD 10 times that of WT enzyme. This study indicates that a normal alpha subunit is required for catalytic activity and alpha-Arg88 likely participates in the isocitrate site, whereas the beta and gamma subunits have roles in the nucleotide functions of this allosteric enzyme. PMID- 14555659 TI - Structure- and function-based characterization of a new phosphoglycolate phosphatase from Thermoplasma acidophilum. AB - The protein TA0175 has a large number of sequence homologues, most of which are annotated as unknown and a few as belonging to the haloacid dehalogenase superfamily, but has no known biological function. Using a combination of amino acid sequence analysis, three-dimensional crystal structure information, and kinetic analysis, we have characterized TA0175 as phosphoglycolate phosphatase from Thermoplasma acidophilum. The crystal structure of TA0175 revealed two distinct domains, a larger core domain and a smaller cap domain. The large domain is composed of a centrally located five-stranded parallel beta-sheet with strand order S10, S9, S8, S1, S2 and a small beta-hairpin, strands S3 and S4. This central sheet is flanked by a set of three alpha-helices on one side and two helices on the other. The smaller domain is composed of an open faced beta sandwich represented by three antiparallel beta-strands, S5, S6, and S7, flanked by two oppositely oriented alpha-helices, H3 and H4. The topology of the large domain is conserved; however, structural variation is observed in the smaller domain among the different functional classes of the haloacid dehalogenase superfamily. Enzymatic assays on TA0175 revealed that this enzyme catalyzed the dephosphorylation of phosphoglycolate in vitro with similar kinetic properties seen for eukaryotic phosphoglycolate phosphatase. Activation by divalent cations, especially Mg2+, and competitive inhibition behavior with Cl- ions are similar between TA0175 and phosphoglycolate phosphatase. The experimental evidence presented for TA0175 is indicative of phosphoglycolate phosphatase. PMID- 14555660 TI - Microsomal prostaglandin E2 synthase-1 is induced by conditional expression of RET/PTC in thyroid PCCL3 cells through the activation of the MEK-ERK pathway. AB - RET/PTC rearrangements are believed to be tumor-initiating events in papillary thyroid carcinomas. We identified microsomal prostaglandin E2 synthase-1 (mPGES 1) as a RET/PTC-inducible gene through subtraction hybridization cloning and expression profiling with custom microarrays. The inducible prostaglandin E2 (PGE2) biosynthetic enzymes cyclooxygenase-2 (COX-2) and mPGES-1 are up-regulated in many cancers. COX-2 is overexpressed in thyroid malignancies compared with benign nodules and normal thyroid tissues. Eicosanoids may promote tumorigenesis through effects on tumor cell growth, immune surveillance, and angiogenesis. Conditional RET/PTC1 or RET/PTC3 expression in PCCL3 thyroid cells markedly induced mPGES-1 and COX-2. PGE2 was the principal prostanoid and up-regulated (by approximately 60-fold), whereas hydroxyeicosatetraenoic acid metabolites were decreased, consistent with shunting of prostanoid biosynthesis toward PGE2 by coactivation of the two enzymes. RET/PTC activated mPGES-1 gene transcription. Based on experiments with kinase inhibitors, with PCCL3 cell lines with doxycycline-inducible expression of RET/PTC mutants with substitutions of critical tyrosine residues in the kinase domain, and lines with inducible expression of activated mutants of H-RAS and MEK1, RET/PTC was found to regulate mPGES-1 through Shc-RAS-MEK-ERK. These data show a direct relationship between activation of a tyrosine kinase receptor oncogene and regulation of PGE2 biosynthesis. As enzymes involved in prostanoid biosynthesis can be targeted with pharmacological inhibitors, these findings may have therapeutic implications. PMID- 14555661 TI - Defective p53 post-translational modification required for wild type p53 inactivation in malignant epithelial cells with mdm2 gene amplification. AB - Mdm2 gene amplification occurs in benign and chemotherapy-responsive malignant tumors with wtp53 genes as well as in breast and epithelial cancers. Mdm2 amplification in benign tumors suggests that it is not sufficient for p53 inactivation in cancer, implying that other defects in the p53 pathway are required for malignancy. We investigated mechanisms of wtp53 protein inactivation in malignant conversion of epithelial cells by comparing clonally related initiated cells with their derivative cancerous cells that have mdm2 amplification. Deficiencies in p53 accumulation and activities in response to DNA damage were not due simply to Mdm2 destabilization of p53 protein, but to continued association of DNA-bound p53 with Mdm2 protein and lack of binding and acetylation by p300 protein. The aberrant interactions were not because of mdm2 amplification alone, because DNA-bound p53 protein from initiated cells failed to bind ectopically expressed Mdm2 or endogenous overexpressed Mdm2 from cancerous cells. Phosphorylations of endogenous p53 at Ser18, -23, or -37 were insufficient to dissociate Mdm2, because each was induced by UV in cancerous cells. Interestingly, phospho-mimic p53-T21E did dissociate the Mdm2 protein from DNA bound p53 and recovered p300 binding and p21 induction in the cancerous cells. Thus wtp53 in malignant cells with mdm2 amplification can be inactivated by continued association of DNA-bound p53 protein with Mdm2 and failure of p300 binding and acetylation, coupled with a defect in p53 phosphorylation at Thr21. These findings suggest therapeutic strategies that address both p53/Mdm2 interaction and associated p53 protein defects in human tumors that have amplified mdm2 genes. PMID- 14555662 TI - Upper airway surface tension: is it a significant cause of airflow obstruction during sleep? PMID- 14555663 TI - Effect of long-term intermittent and sustained hypoxia on hypoxic ventilatory and metabolic responses in the adult rat. AB - The effects of chronic sustained hypoxia (SH) on ventilation have been thoroughly studied. However, the effects of intermittent hypoxia (IH), a more prevalent condition in health and disease are currently unknown. We hypothesized that the ventilatory consequences of SH and IH may differ and be related to changes in N methyl-D-aspartate (NMDA) glutamate receptor subunit expression. To examine these issues, Sprague-Dawley adult male rats were exposed to 30 days of either SH (10% O2) or IH (21% and 10% O2 alternations every 90 s) or to normoxia (RA), at the end of which ventilatory and O2 consumption responses to a 20-min acute hypoxic challenge (10% O2) were conducted. In addition, dorsocaudal brain stem tissue lysates were harvested at 1 h, 6 h, 1 day, 3 days, 7 days, 14 days, and 30 days of SH and IH and analyzed for NR1, NR2A, and NR2B NMDA glutamate receptor expression by immunoblotting. Normoxic ventilation was higher after both SH and IH (P < 0.001). Peak hypoxic ventilatory response was higher after SH but not after IH compared with RA. However, hypoxic ventilatory decline was more prominent after SH than IH (P < 0.001). NR1 expression showed a biphasic pattern of expression over time that was essentially identical after IH and SH (P value not significant). However, NR2A and NR2B expression was higher in IH compared with SH and RA (P < 0.01). We conclude that long-lasting exposures to SH and IH enhance normoxic ventilation but are associated with different time domains of ventilation during acute hypoxia that may be accounted in part by changes in NMDA glutamate receptor subunit expression. PMID- 14555664 TI - Altered ion transporter expression in bronchial epithelium in mountaineers with high-altitude pulmonary edema. AB - Hypoxia inhibits activity and expression of transport proteins of cultured lung alveolar epithelial cells. Here we tested whether hypoxia at high altitude affected the expression of ion transport proteins in tissues obtained from controls and mountaineers with high-altitude pulmonary edema (HAPE) at the Capanna Margherita (4,559 m). Expression was determined by RT-PCR and Western blots from brush biopsies of bronchial epithelium and from leukocytes obtained before and during the stay at high altitude. At low altitude, amounts of mRNAs were not different between control and HAPE-susceptible subjects. At high altitude, the amount of mRNA of Na-K-ATPase, CFTR, and beta-actin of brush biopsies did not change in controls but decreased significantly (-60%) in HAPE susceptible subjects. There was no change in Na channel mRNAs at high altitude in controls and HAPE. No statistically significant correlation was found between the expression of Na transporters and PO2 and O2 saturation. In leukocytes, 28S-rRNA and Na-K-ATPase decreased at altitude in control and HAPE-susceptible subjects, but no significant change in Na-K-ATPase protein was found. Hypoxia-inducible factor-1alpha mRNA and GAPDH mRNA tended to increase in leukocytes obtained from HAPE-susceptible subjects at high altitude but did not change in controls. These results show that hypoxia induces differences in mRNA expression of ion transport related proteins between HAPE-susceptible and control subjects but that these changes may not necessarily predict differences in protein concentration or activity. It is therefore unclear whether these differences are related to the pathophysiology of HAPE. PMID- 14555665 TI - Age-associated changes in skeletal muscles and their effect on mobility: an operational diagnosis of sarcopenia. AB - Sarcopenia, the reduction of muscle mass and strength that occurs with aging, is widely considered one of the major causes of disability in older persons. Surprisingly, criteria that may help a clinician to identify persons with impaired muscle function are still lacking. Using data from a large representative sample of the general population, we examined how muscle function and calf muscle area change with aging and affect mobility in men and women free of neurological conditions. We tested several putative indicators of sarcopenia, including knee extension isometric torque, handgrip, lower extremity muscle power, and calf muscle area. For each indicator, sarcopenia was considered to be present when the measure was >2 SDs below the mean. For all four measures, the prevalence of sarcopenia increased with age, both in men and women. The age associated gradient in prevalence was maximum for muscle power and minimum for calf-muscle area. However, lower extremity muscle power was no better than knee extension torque or handgrip in the early identification of poor mobility, defined either as walking speed <0.8 m/s or inability to walk at least 1 km without difficulty and without developing symptoms. Optimal cutoff values that can be used in the clinical practice to identify older persons with poor mobility were developed. The findings of the study lay the basis for a cost-effective, clinical marker of sarcopenia based on a measure of isometric handgrip strength. Our findings should be verified in a longitudinal study. PMID- 14555666 TI - Acute pathophysiological effects of muscle-expressed Dp71 transgene on normal and dystrophic mouse muscle. AB - products of the dystrophin gene range from the 427-kDa full-length dystrophin to the 70.8-kDa Dp71. Dp427 is expressed in skeletal muscle, where it links the actin cytoskeleton with the extracellular matrix via a complex of dystrophin associated proteins (DAPs). Dystrophin deficiency disrupts the DAP complex and causes muscular dystrophy in humans and the mdx mouse. Dp71, the major nonmuscle product, consists of the COOH-terminal part of dystrophin, including the binding site for the DAP complex but lacks binding sites for microfilaments. Dp71 transgene (Dp71tg) expressed in mdx muscle restores the DAP complex but does not prevent muscle degeneration. In wild-type (WT) mouse muscle, Dp71tg causes a mild muscular dystrophy. In this study, we tested, using isolated extensor digitorum longus muscles, whether Dp71tg exerts acute influences on force generation and sarcolemmal stress resistance. In WT muscles, there was no effect on isometric twitch and tetanic force generation, but with a cytomegalovirus promotor-driven transgene, contraction with stretch led to sarcolemmal ruptures and irreversible loss of tension. In MDX muscle, Dp71tg reduced twitch and tetanic tension but did not aggravate sarcolemmal fragility. The adverse effects of Dp71 in muscle are probably due to its competition with dystrophin and utrophin (in MDX muscle) for binding to the DAP complex. PMID- 14555667 TI - Respiratory effects in humans of a 5-day elevation of end-tidal PCO2 by 8 Torr. AB - The aims of this study were to determine 1) whether ventilatory adaptation occurred over a 5-day exposure to a constant elevation in end-tidal PCO2 and 2) whether such an exposure altered the sensitivity of the chemoreflexes to acute hypoxia and hypercapnia. Ten healthy human subjects were studied over a period of 13 days. Their ventilation, chemoreflex sensitivities, and acid-base status were measured daily before, during, and after 5 days of elevated end-tidal PCO2 at 8 Torr above normal. There was no major adaptation of ventilation during the 5 days of hypercapnic exposure. There was an increase in ventilatory chemosensitivity to acute hypoxia (from 1.35 +/- 0.08 to 1.70 +/- 0.07 l/min/%; P < 0.01) but no change in ventilatory chemosensitivity to acute hypercapnia. There was a degree of compensatory metabolic alkalosis. The results do not support the hypothesis that the ventilatory adaptation to chronic hypercapnia would be much greater with constant elevation of alveolar PCO2 than with constant elevation of inspired PCO2, as has been used in previous studies and in which the feedback loop between ventilation and alveolar PCO2 is left intact. PMID- 14555668 TI - High-speed running performance: a new approach to assessment and prediction. AB - We hypothesized that all-out running speeds for efforts lasting from a few seconds to several minutes could be accurately predicted from two measurements: the maximum respective speeds supported by the anaerobic and aerobic powers of the runner. To evaluate our hypothesis, we recruited seven competitive runners of different event specialties and tested them during treadmill and overground running on level surfaces. The maximum speed supported by anaerobic power was determined from the fastest speed that subjects could attain for a burst of eight steps (approximately 3 s or less). The maximum speed supported by aerobic power, or the velocity at maximal oxygen uptake, was determined from a progressive, discontinuous treadmill test to failure. All-out running speeds for trials of 3 240 s were measured during 10-13 constant-speed treadmill runs to failure and 4 track runs at specified distances. Measured values of the maximum speeds supported by anaerobic and aerobic power, in conjunction with an exponential constant, allowed us to predict the speeds of all-out treadmill trials to within an average of 2.5% (R2 = 0.94; n = 84) and track trials to within 3.4% (R2 = 0.86; n = 28). An algorithm using this exponent and only two of the all-out treadmill runs to predict the remaining treadmill trials was nearly as accurate (average = 3.7%; R2 = 0.93; n = 77). We conclude that our technique 1) provides accurate predictions of high-speed running performance in trained runners and 2) offers a performance assessment alternative to existing tests of anaerobic power and capacity. PMID- 14555669 TI - Upper airway response to electrical stimulation of the genioglossus in obstructive sleep apnea. AB - Contraction of the genioglossus (GG) has been shown to improve upper airway patency. In the present study, we evaluated responses in upper airway pressure flow relationships during sleep to electrical stimulation (ES) of the GG in patients with obstructive sleep apnea. Five patients with chronically implanted hypoglossal nerve (HG) electrodes and nine patients with fine-wire electrodes inserted into the GG were studied. Airflow was measured at multiple levels of nasal pressure, and upper airway collapsibility was defined by the nasal pressure below which airflow ceased ["critical" pressure (Pcrit)]. ES shifted the pressure flow relationships toward higher flow levels in all patients over the entire range of nasal pressure applied. Pcrit decreased similarly during both HG-ES and GG-ES (deltaPcrit was 3.98 +/- 2.31 and 3.18 +/- 1.70 cmH2O, respectively) without a significant change in upstream resistance. The site of collapse (velo- vs. oropharynx) did not influence the response to GG-ES. Moreover, ES-induced reductions in the apnea-hypopnea index of the HG-ES patients were associated with substantial decreases in Pcrit. Our findings imply that responses in apnea severity to HG-ES can be predicted by characterizing the patient's baseline pressure-flow relationships and response to GG-ES. PMID- 14555670 TI - Comparison of body composition methods in overweight and obese children. AB - The objective of the present study was to investigate the accuracy of percent body fat (%fat) estimates from dual-energy X-ray absorptiometry, air-displacement plethysmography (ADP), and total body water (TBW) against a criterion four compartment (4C) model in overweight and obese children. A volunteer sample of 30 children (18 male and 12 female), age of (mean +/- SD) 14.10 +/- 1.83 yr, body mass index of 31.6 +/- 5.5 kg/m, and %fat (4C model) of 41.2 +/- 8.2%, was assessed. Body density measurements were converted to %fat estimates by using the general equation of Siri (ADPSiri) (Siri WE. Techniques for Measuring Body Composition. 1961) and the age- and gender-specific constants of Lohman (ADPLoh) (Lohman TG. Exercise and Sport Sciences Reviews. 1986). TBW measurements were converted to %fat estimates by assuming that water accounts for 73% of fat-free mass (TBW73) and by utilizing the age- and gender-specific water contents of Lohman (TBWLoh). All estimates of %fat were highly correlated with those of the 4C model (r > or = 0.95, P < 0.001; SE < or = 2.14). For %fat, the total error and mean difference +/- 95% limits of agreement compared with the 4C model were 2.50, 1.8 +/- 3.5 (ADPSiri); 1.82, -0.04 +/- 3.6 (ADPLoh); 2.86, -2.0 +/- 4.1 (TBW73); 1.90, -0.3 +/- 3.8 (TBWLoh); and 2.74, 1.9 +/- 4.0 DXA (dual-energy X ray absorptiometry), respectively. In conclusion, in overweight and obese children, ADPLoh and TBWLoh were the most accurate methods of measuring %fat compared with a 4C model. However, all methods under consideration produced similar limits of agreement. PMID- 14555671 TI - Effect of hyperbaric oxygen on apoptosis in neonatal hypoxia-ischemia rat model. AB - We have previously demonstrated that a transient exposure to hyperbaric oxygen (HBO) attenuated the neuronal injury after neonatal hypoxia-ischemia. This study was undertaken to determine whether HBO offers this neuroprotection by reducing apoptosis in injured brain tissue. Seven-day-old rat pups were subjected to unilateral carotid artery ligation followed by 2 h of hypoxia (8% oxygen). Apoptotic cell death was examined in the injured cortex and hippocampus tissue. Caspase-3 expression and activity increased at 18 and 24 h after the hypoxia ischemia insult. At 18-48 h, poly(ADP-ribose) polymerase (PARP) cleavage occurred, which reduced the band at 116 kDa and enhanced the band at 85 kDa. There was a time-dependent increase in the number of terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL)-positive cells. A single HBO treatment (100% oxygen, 3 ATA for 1 h) 1 h after hypoxia reduced the enhanced caspase-3 expression and activity, attenuated the PARP cleavage, and decreased the number of TUNEL-positive cells observed in the cortex and hippocampus. These results suggest that the neuroprotective effect of HBO is at least partially mediated by the reduction of apoptosis. PMID- 14555672 TI - Age-related changes in the mitotic and metabolic characteristics of muscle derived cells. AB - Age-related sarcopenia could partly result from cumulative repeated episodes of incomplete repair and regeneration. We hypothesized that mitotic and metabolic events associated with satellite cell activation and proliferation could be altered with aging. Muscle-derived cells (mdc) were isolated from gastrocnemius and quadriceps muscles of young (3 wk old), adult (9 mo old), and old (24 mo old) Sprague-Dawley male rats (n = 10/group). The mdc from young growing rats started to proliferate earlier compared with adult and old animals. Cell cycle duration was significantly reduced with aging from 36.5 +/- 3.2 to 28.0 +/- 2.2 h. However, the proportion of noncycling (G0 phase) and cycling (G1 + S + G2 + M phases) cultured mdc was statistically unchanged among the three age groups. Significantly lower increase in c-met and proliferating cell nuclear antigen expression were observed in cultured mdc of old rats upon serum stimulation. Major changes in the expression of citrate synthase, lactate dehydrogenase, proteasome, caspase 3, plasminogen activators (PAs), and matrix metalloproteinase 2-9 (MMP2-9) were observed upon serum stimulation, but no age-related difference was noted. However, when measured on crushed muscle extracts, PAs and MMP2-9 enzyme activities were significantly decreased with aging. Our results show that cellular and biochemical events associated with the control of mdc activation and proliferation occur with aging. These alterations may participate in the accumulation of repeated episodes of incomplete repair and regeneration throughout the life span, thus contributing to the loss of skeletal muscle mass and function with aging. PMID- 14555673 TI - Preventing annoyance from odors in spaceflight: a method for evaluating the sensory impact of rodent housing. AB - For the scientific community, the ability to fly mice under weightless conditions in space offers several advantages over the use of rats. These advantages include the option of testing a range of transgenic animals, the ability to increase the number of animals that can be flown, and reduced demands on shuttle resources (food, water, animal mass) and crew time (for water refill). Mice have been flown in animal enclosure module (AEM) hardware only once [Space Shuttle Transport System (STS)-90] and were dissected early in the mission, whereas rats have been flown in the AEM on >20 missions. This has been due, in part, to concerns that strong and annoying odors from mouse urine (vs. rat urine) will interfere with crew performance in the shuttle middeck. To screen and approve mice for flight, a method was developed to evaluate the odor containment performance of AEMs housing female C57BL/6J mice compared with AEMs housing Sprague-Dawley rats across a 21 day test period. Based on the results of this test, consensus was reached that mice could fly in the AEM hardware for up to 17 days (including prelaunch and contingency) and that the AEM hardware would likely contain odors beyond this duration. Human sensory and electronic nose analysis of the AEMs postflight demonstrated their success in containing odors from mice for the mission duration of STS-108 (13 days). Although this paper focuses specifically on odor evaluations for the space shuttle, the concern is applicable to any confined, closed-system environment for human habitation. PMID- 14555674 TI - Invited Review: Role of mechanophysiology in aging of ECM: effects of changes in mechanochemical transduction. AB - Mechanical forces play a role in the development and evolution of extracellular matrices (ECMs) found in connective tissue. Gravitational forces acting on mammalian tissues increase the net muscle forces required for movement of vertebrates. As body mass increases during development, musculoskeletal tissues and other ECMs are able to adapt their size to meet the increased mechanical requirements. However, the control mechanisms that allow for rapid growth in tissue size during development are altered during maturation and aging. The purpose of this mini-review is to examine the relationship between mechanical loading and cellular events that are associated with downregulation of mechanochemical transduction, which appears to contribute to aging of connective tissue. These changes result from decreases in growth factor and hormone levels, as well as decreased activation of the phosphorelay system that controls cell division, gene expression, and protein synthesis. Studies pertaining to the interactions among mechanical forces, growth factors, hormones, and their receptors will better define the relationship between mechanochemical transduction processes and cellular behavior in aging tissues. PMID- 14555675 TI - Invited Review: Pathogenesis of osteoporosis. AB - Patients with fragility fractures may have abnormalities in bone structural and material properties such as larger or smaller bone size, fewer and thinner trabeculae, thinned and porous cortices, and tissue mineral content that is either too high or too low. Bone models and remodels throughout life; however, with advancing age, less bone is replaced than was resorbed within each remodeling site. Estrogen deficiency at menopause increases remodeling intensity: a greater proportion of bone is remodeled on its endosteal (inner) surface, and within each of the many sites even more bone is lost as more bone is resorbed while less is replaced, accelerating architectural decay. In men, there is no midlife increase in remodeling. Bone loss within each remodeling site proceeds by reduced bone formation, producing trabecular and cortical thinning. Hypogonadism in 20-30% of elderly men contributes to bone loss. In both sexes, calcium malabsorption and secondary hyperparathyroidism increase remodeling: more bone is removed from an ever-diminishing bone mass. As bone is removed from the endosteal envelope, concurrent bone formation on the periosteal (outer) bone surface during aging partly offsets bone loss and increases bone's cross-sectional area. Periosteal apposition is less in women than in men; therefore, women have more net bone loss because they gain less on the periosteal surface, not because they resorb more on the endosteal surface. More women than men experience fractures because their smaller skeleton incurs greater architectural damage and adapts less by periosteal apposition. PMID- 14555676 TI - Invited Review: Dynamic exercise performance in Masters athletes: insight into the effects of primary human aging on physiological functional capacity. AB - Physiological functional capacity (PFC) is defined here as the ability to perform the physical tasks of daily life and the ease with which these tasks can be performed. For the past decade, we have sought to determine the effect of primary (healthy) adult human aging on PFC and the potential modulatory influences of gender and habitual aerobic exercise status on this process by studying young adult and Masters athletes. An initial approach to determining the effects of aging on PFC involved investigating changes in peak exercise performance with age in highly trained and competitive athletes. PFC, as assessed by running and swimming performance, decreased only modestly until age 60-70 yr but declined exponentially thereafter. A progressive reduction in maximal O2 consumption (V(O2 max)) appears to be the primary physiological mechanism associated with declines in endurance running performance with advancing age, along with a reduction in the exercise velocity at lactate threshold. Because V(O2 max) is important in mediating age-related reductions in exercise performance and PFC, we then investigated the modulatory influence of habitual aerobic exercise status on the rate of decline in V(O2 max) with age. Surprisingly, as a group, endurance trained adults appear to undergo greater absolute rates of decline in V(O2 max) with advancing age compared with healthy sedentary adults. This appears to be mediated by a baseline effect (higher V(O2 max) as young adults) and/or a marked age-related decline in exercise training volume and intensity (stimulus) in endurance-trained adults. Thus the ability to maintain habitual physical activity levels with advancing age appears to be a critical determinant of changes in PFC in part via modulation of maximal aerobic capacity. PMID- 14555678 TI - HMB meta-analysis and the clustering of data sources. PMID- 14555679 TI - Modification of a sigmoidal equation for the pulmonary pressure-volume curve for asymmetric data. PMID- 14555680 TI - Impact of muscle length during stretch-shortening contractions on real-time and temporal muscle performance measures in rats in vivo. AB - The objective of the present study was to investigate the impact of muscle length during stretch-shortening cycles on static and dynamic muscle performance. Animals were randomly assigned to an isometric (control, Con, n = 12), a short muscle-length (S-Inj, 1.22-2.09 rad, n = 12), or a long-muscle-length (L-Inj, 1.57-2.44 rad, n = 12) group. The dorsiflexor muscles were exposed in vivo to 7 sets of 10 stretch-shortening contractions (conducted at 8.72 rad/s) or 7 sets of isometric contractions of the same stimulation duration by using a custom designed dynamometer. Performance was characterized by multipositional isometric exertions and positive, negative, and net work before exposure, 6 h after exposure, and 48 h after exposure to contractions. Real-time muscle performance during the stretch-shortening cycles was characterized by stretch-shortening parameters and negative, positive, and net work. The S-Inj group recovery (force difference) was similar to the Con group force difference at 48 h, whereas the L Inj group force difference was statistically greater at 1.39, 1.57, and 1.74 rad than the Con group force difference (P < 0.05). Negative work (P < 0.05) and net work (P < 0.05) were statistically lower in the S-Inj and L-Inj groups than in the Con group 48 h after exposure to contractions. Of the real-time parameters, there was a difference in cyclic force with treatment during the stretch shortening cycles (P < 0.0001), with the L-Inj group being the most affected. Thus longer ranges of motion result in a more profound isometric force decrement 48 h after exposure to contractions and in real-time changes in eccentric forces. PMID- 14555681 TI - Nitric oxide-cGMP pathway is involved in endotoxin-induced contractile dysfunction in rat hearts. AB - The mechanisms by which endotoxemia causes cardiac depression have not been fully elucidated. The present study examined the involvement of nitric oxide (NO) in this pathology. Rats were infused with lipopolysaccharide (LPS) or saline, and the plasma and myocardial NO(2)(-) and NO(3)(-) (NOx) concentrations were measured before or 3, 6, and 24 h after treatment. The hearts were then immediately isolated and mounted in a Langendorff apparatus, and left ventricular developed pressure (LVDP) was determined before biochemical analysis of the myocardium. LPS injection effected the expression of inducible NO synthase (iNOS) in the myocardium, a marked increase in plasma and myocardial NOx levels, and a significant decline in LVDP compared with saline controls. The LPS-induced NO production and concomitant cardiac depression were most pronounced 6 h after LPS injection and were accompanied by a significant increase in myocardial cGMP content. Myocardial ATP levels were not significantly altered after LPS injection. Significant negative correlation was observed between LVDP and myocardial cGMP content, as well as between LVDP and plasma NOx levels. Aminoguanidine, an inhibitor of iNOS, significantly attenuated the LPS-induced NOx production and contractile dysfunction. Furthermore, 1H-[1,2,4]oxadiazolo[4,3 a]quinoxalin-1-one, an inhibitor of soluble guanylate cyclase, significantly decreased myocardial cGMP content and attenuated the contractile depression, although aminoguanidine or 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one was not able to completely reverse myocardial dysfunction. Our data suggest that endotoxin-induced contractile dysfunction in rat hearts is associated with NO production by myocardial iNOS and a concomitant increase in myocardial cGMP. PMID- 14555682 TI - Arm-cranking muscle power and arm isometric muscle strength are independent predictors of all-cause mortality in men. AB - Poor muscle strength is associated with mortality, presumably due to low muscle mass. Notably, muscle power declines more rapidly than muscle strength with increasing age, which may be related to more complex central nervous system movement control. We examined arm-cranking power against four workloads and isometric strength measured in the upper extremities of 993 men longitudinally tested over a 25-yr period. Muscle mass was estimated by using 24-h creatinine excretion; physical activity was assessed by self-reported questionnaire. Muscle power and strength were modeled by time by using mixed-effects models, which developed regression equations for each individual. The first derivative of these equations estimated rate of change in strength or power at each evaluation. Survival analyses, using the counting method, examined the impact of strength, power, and their rates of change on all-cause mortality while adjusting for age. Arm-cranking power [relative risk (rr) = 0.984 per 100 kg.m.min(-1), P < 0.001] was a stronger predictor of mortality than was arm strength (rr = 0.986 per 10 kg, P = not significant), whereas rate of power change (rr = 0.989 per 100 kg.min(-1).yr(-1)) and rate of arm strength change (rr = 0.888 per 10 kg/yr) were risks independent of the power or strength levels. The impacts of power and strength were partially independent of muscle mass and physical activity. The risk of mortality was similar across the four power workloads (rr = 0.93-0.96 per 100 kg.m.min(-1)), whereas the lowest load generated less than one-half the power as the higher loads. Arm-cranking power is a risk factor for mortality, independent of muscle strength, physical activity, and muscle mass. The impact is found with loads that do not generate maximal power, suggesting an important role for motor coordination and speed of movement. PMID- 14555683 TI - Hypoxia-mediated prolonged elevation of sympathetic nerve activity after periods of intermittent hypoxic apnea. AB - Obstructive sleep apnea (OSA) is associated with transient elevation of muscle sympathetic nerve activity (MSNA) during apneic events, which often produces elevated daytime MSNA in OSA patients. Hypoxia is postulated to be the primary stimulus for elevated daytime MSNA in OSA patients. Therefore, we studied the effects of 20 min of intermittent voluntary hypoxic apneas on MSNA during 180 min of recovery. Also, we compared MSNA during recovery after either 20 min of intermittent voluntary hypoxic apneas, hypercapnic hypoxia, or isocapnic hypoxia. Consistent with our hypothesis, both total MSNA and MSNA burst frequency were elevated after 20 min of intermittent hypoxic apnea compared with baseline (P < 0.05). Both total MSNA and MSNA burst frequency remained elevated throughout the 180-min recovery period and were statistically different from time control subjects throughout this period (P < 0.05). Finally, MSNA during recovery from intermittent hypoxic apnea, hypercapnic hypoxia, and isocapnic hypoxia were not different (P = 0.50). Therefore, these data support the hypothesis that short term exposure to intermittent hypoxic apnea results in sustained elevation of MSNA and that hypoxia is the primary mediator of this response. PMID- 14555684 TI - Differential effects of sodium selenite in reducing tissue damage caused by three hemoglobin-based oxygen carriers. AB - Three "blood substitutes," a diaspirin cross-linked human hemoglobin (DBBF-Hb), a bovine polymerized hemoglobin (PolyHbBv), and a human polymerized hemoglobin (O-R PolyHbA(0)), that have undergone clinical trials are used in this study. Previously, we showed in the rat that coadministration of sodium selenite (Na(2)SeO(3)) and DBBF-Hb significantly decreased mesenteric venular leakage and epithelial disruption produced by DBBF-Hb alone but did not reduce mast cell degranulation unless given orally. The purpose of this study was to determine whether Na(2)SeO(3) produced similar beneficial responses when used with PolyHbBv and O-R-PolyHbA(0). In anesthetized Sprague-Dawley rats, the mesenteric microvasculature was perfused with PolyHbBv or O-R-PolyHbA(0), with and without Na(2)SeO(3) in the perfusate and suffusate, for 10 min, followed by FITC-albumin for 3 min, and then fixed for microscopy. Na(2)SeO(3) did not reduce leak number or area in preparations perfused with PolyHbBv and only reduced leak number (but not significantly) in preparations perfused with O-R-PolyHbA(0). Na(2)SeO(3) significantly increased mesenteric mast cell degranulation and impaired epithelial integrity in animals treated with PolyHbBv. In vitro, Na(2)SeO(3) significantly reduced the oxidation rate of DBBF-Hb in the presence of oxidants, had little effect on PolyHbBv, and increased the oxidation rate of O-R PolyHbA(0). These results suggest that Na(2)SeO(3) moderates hemoglobin-induced damage, at least partly, through its redox interactions with the heme sites in the hemoglobin molecules studied and that accessibility of the heme site to Na(2)SeO(3) governs those interactions. PMID- 14555686 TI - Role of insulin on exercise-induced GLUT-4 protein expression and glycogen supercompensation in rat skeletal muscle. AB - The purpose of this study was to investigate the role of insulin on skeletal muscle GLUT-4 protein expression and glycogen storage after postexercise carbohydrate supplementation. Male Sprague-Dawley rats were randomly assigned to one of six treatment groups: sedentary control (Con), Con with streptozocin (Stz/C), immediately postexercise (Ex0), Ex0 with Stz (Stz/Ex0), 5-h postexercise (Ex5), and Ex5 with Stz (Stz/Ex5). Rats were exercised by swimming (2 bouts of 3 h) and carbohydrate supplemented immediately after each exercise session by glucose intubation (1 ml of a 50% wt/vol). Stz was administered 72-h before exercise, which resulted in hyperglycemia and elimination of the insulin response to the carbohydrate supplement. GLUT-4 protein of Ex0 rats was 30% above Con in fast-twitch (FT) red and 21% above Con in FT white muscle. In Ex5, GLUT-4 protein was 52% above Con in FT red and 47% above Con in FT white muscle. Muscle glycogen in FT red and white muscle was also increased above Con in Ex5 rats. Neither GLUT 4 protein nor muscle glycogen was increased above Con in Stz/Ex0 or Stz/Ex5 rats. GLUT-4 mRNA in FT red muscle of Ex0 rats was 61% above Con but only 33% above Con in Ex5 rats. GLUT-4 mRNA in FT red muscle of Stz/C and Stz/Ex0 rats was similar but significantly elevated in Ex5/Stz rats. These results suggest that insulin is essential for the increase in GLUT-4 protein expression following postexercise carbohydrate supplementation. PMID- 14555685 TI - HSP90 and Akt modulate Ang-1-induced angiogenesis via NO in coronary artery endothelium. AB - This study examines the notion that heat shock protein (HSP) 90 binding to nitric oxide (NO), endothelial NO synthase (eNOS), and PI3K-Akt regulate angiopoietin (Ang)-1-induced angiogenesis in porcine coronary artery endothelial cells (PCAEC). Exposure to Ang-1 (250 ng/ml) for periods up to 2 h resulted in a time dependent increase in eNOS phosphorylation at Ser 1177 that occurred by 5 min and peaked at 60 min. This was accompanied by a gradual increase in NO release. Ang-1 also led to stimulation of HSP90 binding to eNOS and a significant increase in Akt phosphorylation. Thirty minutes of pretreatment of cells with either 1 microg/ml geldanamycin (a specific inhibitor of HSP90) or 500 nM wortmannin [a specific phosphatidylinositol 3 (PI3)-kinase (PI3K) inhibitor] significantly attenuated Ang-1-stimulated eNOS phosphorylation and NO production. Exposure to Ang-1 caused an increase in endothelial cell migration, tube formation, and sprouting from PCAEC spheroids, and pharmacological blockage of HSP90 function or inhibition of PI3K-Akt pathway completely abolished these effects. Inhibition of nitric oxide synthase by NG-nitro-l-arginine methyl ester (2.5 mM) also resulted in a significant decrease in Ang-1-induced angiogenesis. We conclude that stimulated HSP90 binding to eNOS and activation of the PI3-Akt pathway contribute to Ang-1-induced eNOS phosphorylation, NO production, and angiogenesis in PCAEC. PMID- 14555687 TI - Postcontraction insulin sensitivity: relationship with contraction protocol, glycogen concentration, and 5' AMP-activated protein kinase phosphorylation. AB - Exercise enhances insulin-stimulated glucose transport (GT) in skeletal muscle. Evidence suggests that 5' AMP-activated protein kinase (AMPK) and glycogen may be important for enhanced insulin sensitivity. Our goals were to investigate the effect of various in situ muscle contraction protocols on insulin-stimulated GT and assess the relationship of contraction-induced changes in AMPK and glycogen with postcontraction improvement in insulin-stimulated GT. Rats were anesthetized, both ulnar nerves were exposed, and one nerve was electrically stimulated to contract forelimb muscles. We performed a series of five experiments, sequentially varying only one contraction parameter (train duration, train rate, pulse frequency, number of 5-min bouts, or pulse duration) while holding the others constant. Both epitrochlearis muscles were dissected out and incubated for 3.5 h before measurement of GT. For each contraction parameter studied, we identified an apparent threshold value that did not induce a significant increase in insulin-stimulated GT and an apparent peak value, above which there was a plateau or decline in insulin-stimulated GT. Using other rats, we evaluated muscle AMPK phosphorylation and glycogen concentration immediately postcontraction. AMPK phosphorylation and reduction in glycogen were increased compared with resting controls in each protocol, which had previously been shown to increase insulin-stimulated GT, as well as in several protocols that did not significantly increase insulin-stimulated GT. These data suggest that contraction induced AMPK phosphorylation and decrease in glycogen may be necessary but are not sufficient for the postcontraction increase in insulin-stimulated GT in rat skeletal muscle. PMID- 14555688 TI - Changes in regional cerebral blood flow distribution during postexercise hypotension in humans. AB - This investigation compared patterns of regional cerebral blood flow (rCBF) during exercise recovery both with and without postexercise hypotension (PEH). Eight subjects were studied on 3 days with randomly assigned conditions: 1) after 30 min of rest; 2) after 30 min of moderate exercise (M-Ex) at 60-70% heart rate (HR) reserve during PEH; and 3) after 30 min of light exercise (L-Ex) at 20% HR reserve with no PEH. Data were collected for HR, mean blood pressure (MBP), and ratings of perceived exertion and relaxation, and rCBF was assessed by use of single-photon-emission computed tomography. With the use of ANOVA across conditions, there were differences (P < 0.05; mean +/- SD) from rest during exercise recovery from M-Ex (HR = +12 +/- 3 beats/min; MBP = -9 +/- 2 mmHg), but not from L-Ex (HR = +2 +/- 2 beats/min; MBP = -2 +/- 2 mmHg). After M-Ex, there were decreases (P < 0.05) for the anterior cingulate (-6.7 +/- 2%), right and left inferior thalamus (-10 +/- 3%), right inferior insula (-13 +/- 3%), and left inferior anterior insula (-8 +/- 3%), not observed after L-Ex. There were rCBF decreases for leg sensorimotor regions after both M-Ex (-15 +/- 4%) and L-Ex (-12 +/- 3%) and for the left superior anterior insula (-7 +/- 3% and -6 +/- 3%), respectively. Data show that there are rCBF reductions within specific regions of the insular cortex and anterior cingulate cortex coupled with a postexercise hypotensive response after M-Ex. Findings suggest that these cerebral cortical regions, previously implicated in cardiovascular regulation during exercise, may also be involved in PEH. PMID- 14555689 TI - Echocardiographic assessment of age-associated changes in systolic and diastolic function of the female F344 rat heart. AB - Aging is associated with hypertrophy, dilatation, and fibrosis of the left ventricle (LV) of the heart. Advances in echocardiographic assessment have made it possible to follow changes in cardiac function in a serial, noninvasive manner. The purpose was to determine whether there is echocardiographic evidence of age-associated changes in chamber dimensions and systolic and diastolic properties of the female Fischer 344 (F344) rat heart. On the basis of previous invasive studies, it was predicted that echocardiographic assessment would detect age-associated changes in indexes of systolic and diastolic function. Rats were sedated with 1.5% isoflurane and placed in the supine position. Two-dimensional images and two-dimensionally guided M-mode, Doppler M mode, Doppler tissue, and pulsed-wave Doppler recordings were obtained from the parasternal long axis, parasternal short axis, and/or apical four-chamber views as per convention by using a 15-MHz linear array or 8-MHz phased-array transducer or a GE S10-MHz phased-array transducer. Compared with young adult 4-mo-old rats, there is a significant decrement in the resting systolic function of the LV in 30-mo-old female F344 rats as evidenced by declines in LV ejection fraction (80 +/- 9 vs. 89 +/- 5%; mean +/- SD), fractional shortening (43 +/- 9 vs. 54 +/- 8%) and velocity of circumferential fiber shortening (2.43 +/- 0.53 vs. 2.99 +/- 0.50 circ/s). Evidence for age-associated differences in diastolic function included an increase in isovolumic relaxation time (25.0 +/- 7.6 vs. 17.2 +/- 4.4 ms) and decreases in the tissue Doppler peak E waves at the septal annulus and at the lateral annulus of the mitral valve. The modest changes in systolic and diastolic LV function that occur with advancing age in the female F344 rat are likely to reduce the capacity of the heart to respond to hemodynamic challenges. PMID- 14555690 TI - Activation of a stress-responsive mitogen-activated protein kinase cascade induces the biosynthesis of ethylene in plants. AB - Plants under stress from both biotic and abiotic sources produce increased levels of ethylene, which is perceived by ethylene receptors and triggers cellular responses further downstream. Protein phosphorylation and dephosphorylation were implicated in the regulation of ethylene induction by stresses based on studies using protein kinase and phosphatase inhibitors. However, the kinase(s) involved remains to be determined. Using a conditional gain-of-function transgenic system, we demonstrate that the activation of SIPK, a tobacco mitogen-activated protein kinase (MAPK), by NtMEK2DD, an active mutant of the upstream kinase of SIPK, resulted in a dramatic increase in ethylene production. The increase in ethylene after the activation of SIPK coincided with a dramatic increase in 1 aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS) activity, which was followed by the activation of a subgroup of ACS and ACC oxidase (ACO) genes, suggesting that either the activation of unidentified ACS(s) or post transcriptional regulation is involved. Infection with Tobacco mosaic virus (TMV), which is known to activate the SIPK cascade and induce ethylene biosynthesis, also induced the same ACSs and ACOs. After ethylene production in NtMEK2DD plants, strong activation of ETHYLENE-RESPONSE FACTOR (ERF) genes was observed, similar to the effect in NN tobacco plants infected with TMV. In contrast to previous reports, no major increase in jasmonic acid (JA) and methyl jasmonate (MJ) was detected after the activation of SIPK/WIPK in NtMEK2DD transgenic plants. These results suggest that the induction of ethylene but not JA/MJ is involved in plant defense responses mediated by the NtMEK2-SIPK/WIPK pathway. PMID- 14555691 TI - The TIME FOR COFFEE gene maintains the amplitude and timing of Arabidopsis circadian clocks. AB - Plants synchronize developmental and metabolic processes with the earth's 24-h rotation through the integration of circadian rhythms and responses to light. We characterize the time for coffee (tic) mutant that disrupts circadian gating, photoperiodism, and multiple circadian rhythms, with differential effects among rhythms. TIC is distinct in physiological functions and genetic map position from other rhythm mutants and their homologous loci. Detailed rhythm analysis shows that the chlorophyll a/b-binding protein gene expression rhythm requires TIC function in the mid to late subjective night, when human activity may require coffee, in contrast to the function of EARLY-FLOWERING3 (ELF3) in the late day to early night. tic mutants misexpress genes that are thought to be critical for circadian timing, consistent with our functional analysis. Thus, we identify TIC as a regulator of the clock gene circuit. In contrast to tic and elf3 single mutants, tic elf3 double mutants are completely arrhythmic. Even the robust circadian clock of plants cannot function with defects at two different phases. PMID- 14555692 TI - Proanthocyanidin-accumulating cells in Arabidopsis testa: regulation of differentiation and role in seed development. AB - Anthocyanidin reductase encoded by the BANYULS (BAN) gene is the core enzyme in proanthocyanidin (PA) biosynthesis. Here, we analyzed the developmental mechanisms that regulate the spatiotemporal expression of BAN in the developing Arabidopsis seed coat. PA-accumulating cells were localized histochemically in the inner integument (seed body and micropyle) and pigment strand (chalaza). BAN promoter activity was detected specifically in these cells. Gain-of-function experiments showed that an 86-bp promoter fragment functioned as an enhancer specific for PA-accumulating cells. Mutations in regulatory genes of PA biosynthesis abolished BAN promoter activity (transparent testa2 [tt2], tt8, and transparent testa glabra1 [ttg1]), modified its spatial pattern (tt1 and tt16), or had no influence (ttg2), thus revealing complex regulatory interactions at several developmental levels. Genetic ablation of PA-accumulating cells targeted by the BAN promoter fused to BARNASE led to the formation of normal plants that produced viable yellow seeds. Importantly, these seeds had no obvious defects in endosperm and embryo development. PMID- 14555693 TI - The BOTRYTIS SUSCEPTIBLE1 gene encodes an R2R3MYB transcription factor protein that is required for biotic and abiotic stress responses in Arabidopsis. AB - The molecular and cellular mechanisms involved in plant resistance to the necrotrophic fungal pathogen Botrytis cinerea and their genetic control are poorly understood. Botrytis causes severe disease in a wide range of plant species, both in the field and in postharvest situations, resulting in significant economic losses. We have isolated the BOS1 (BOTRYTIS-SUSCEPTIBLE1) gene of Arabidopsis based on a T-DNA insertion allele that resulted in increased susceptibility to Botrytis infection. The BOS1 gene is required to restrict the spread of another necrotrophic pathogen, Alternaria brassicicola, suggesting a common host response strategy against these pathogens. In the case of the biotrophic pathogens Pseudomonas syringae pv tomato and the oomycete parasite Peronospora parasitica, bos1 exhibits enhanced disease symptoms, but pathogen growth is similar in bos1 and wild-type plants. Strikingly, bos1 plants have impaired tolerance to water deficit, increased salinity, and oxidative stress. Botrytis infection induces the expression of the BOS1 gene. This increased expression is severely impaired in the coi1 mutant, suggesting an interaction of BOS1 with the jasmonate signaling pathway. BOS1 encodes an R2R3MYB transcription factor protein, and our results suggest that it mediates responses to signals, possibly mediated by reactive oxygen intermediates from both biotic and abiotic stress agents. PMID- 14555694 TI - Cytokinin-deficient transgenic Arabidopsis plants show multiple developmental alterations indicating opposite functions of cytokinins in the regulation of shoot and root meristem activity. AB - Cytokinins are hormones that regulate cell division and development. As a result of a lack of specific mutants and biochemical tools, it has not been possible to study the consequences of cytokinin deficiency. Cytokinin-deficient plants are expected to yield information about processes in which cytokinins are limiting and that, therefore, they might regulate. We have engineered transgenic Arabidopsis plants that overexpress individually six different members of the cytokinin oxidase/dehydrogenase (AtCKX) gene family and have undertaken a detailed phenotypic analysis. Transgenic plants had increased cytokinin breakdown (30 to 45% of wild-type cytokinin content) and reduced expression of the cytokinin reporter gene ARR5:GUS (beta-glucuronidase). Cytokinin deficiency resulted in diminished activity of the vegetative and floral shoot apical meristems and leaf primordia, indicating an absolute requirement for the hormone. By contrast, cytokinins are negative regulators of root growth and lateral root formation. We show that the increased growth of the primary root is linked to an enhanced meristematic cell number, suggesting that cytokinins control the exit of cells from the root meristem. Different AtCKX-green fluorescent protein fusion proteins were localized to the vacuoles or the endoplasmic reticulum and possibly to the extracellular space, indicating that subcellular compartmentation plays an important role in cytokinin biology. Analyses of promoter:GUS fusion genes showed differential expression of AtCKX genes during plant development, the activity being confined predominantly to zones of active growth. Our results are consistent with the hypothesis that cytokinins have central, but opposite, regulatory functions in root and shoot meristems and indicate that a fine-tuned control of catabolism plays an important role in ensuring the proper regulation of cytokinin functions. PMID- 14555695 TI - Limitations on geminivirus genome size imposed by plasmodesmata and virus-encoded movement protein: insights into DNA trafficking. AB - Animals and plants evolved systems to permit non-cell-autonomous trafficking of RNA, whereas DNA plays a cell-autonomous role. In plants, plasmodesmata serve as the conduit for this phenomenon, and viruses have evolved to use this pathway for the spread of infectious nucleic acids. In this study, a plant DNA virus was used to explore the constraints imposed on the movement of DNA through this endogenous RNA trafficking pathway. The combined properties of the geminivirus-encoded movement protein and plasmodesmata were shown to impose a strict limitation on the size of the viral genome at the level of cell-to-cell movement. Size increased viral genome components underwent homologous and nonhomologous recombination to overcome this strict limitation. Our results provide insights into the genetic mechanisms that underlie viral evolution and provide a likely explanation for why relatively few types of plant DNA viruses have evolved: they would have had to overcome the constraints imposed by an endogenous system operating to ensure that DNA acts in a cell-autonomous manner. PMID- 14555696 TI - MADS-box protein complexes control carpel and ovule development in Arabidopsis. AB - The AGAMOUS (AG) gene is necessary for stamen and carpel development and is part of a monophyletic clade of MADS-box genes that also includes SHATTERPROOF1 (SHP1), SHP2, and SEEDSTICK (STK). Here, we show that ectopic expression of either the STK or SHP gene is sufficient to induce the transformation of sepals into carpeloid organs bearing ovules. Moreover, the fact that these organ transformations occur when the STK gene is expressed ectopically in ag mutants shows that STK can promote carpel development in the absence of AG activity. We also show that STK, AG, SHP1, and SHP2 can form multimeric complexes and that these interactions require the SEPALLATA (SEP) MADS-box proteins. We provide genetic evidence for this role of the SEP proteins by showing that a reduction in SEP activity leads to the loss of normal ovule development, similar to what occurs in stk shp1 shp2 triple mutants. Together, these results indicate that the SEP proteins, which are known to form multimeric complexes in the control of flower organ identity, also form complexes to control normal ovule development. PMID- 14555697 TI - Two discrete cis elements control the Abaxial side-specific expression of the FILAMENTOUS FLOWER gene in Arabidopsis. AB - Our previous studies showed that a member of the YABBY gene family, FILAMENTOUS FLOWER (FIL), plays a role in specifying the abaxial side tissues in the development of lateral organs such as cotyledons, leaves, young flower buds, and flower organs. We examined the expression pattern of FIL and found a temporal change of expression domains in the developmental process of the floral meristem. We also examined the cis control regions by constructing a series of transgenic plants that carry green fluorescent protein under the control of the FIL promoter with several types of deletions, base changes, and tandem repeats and showed that the unique expression pattern is dependent on at least two cis-acting elements in the 5' regulatory region. One element proximal to the FIL gene would be responsible for the expression of both the abaxial and adaxial sides, and the other element of the 12-bp sequence would work to repress expression on the adaxial side. PMID- 14555698 TI - An Arabidopsis Callose Synthase, GSL5, Is Required for Wound and Papillary Callose Formation. AB - Arabidopsis was transformed with double-stranded RNA interference (dsRNAi) constructs designed to silence three putative callose synthase genes: GLUCAN SYNTHASE-LIKE5 (GSL5), GSL6, and GSL11. Both wound callose and papillary callose were absent in lines transformed with GSL5 dsRNAi and in a corresponding sequence indexed GSL5 T-DNA insertion line but were unaffected in GSL6 and GSL11 dsRNAi lines. These data provide strong genetic evidence that the GSL genes of higher plants encode proteins that are essential for callose formation. Deposition of callosic plugs, or papillae, at sites of fungal penetration is a widely recognized early response of host plants to microbial attack and has been implicated in impeding entry of the fungus. Depletion of callose from papillae in gsl5 plants marginally enhanced the penetration of the grass powdery mildew fungus Blumeria graminis on the nonhost Arabidopsis. Paradoxically, the absence of callose in papillae or haustorial complexes correlated with the effective growth cessation of several normally virulent powdery mildew species and of Peronospora parasitica. PMID- 14555700 TI - Differential subnuclear localization of RNA strands of opposite polarity derived from an autonomously replicating viroid. AB - The wide variety of RNAs produced in the nucleus must be localized correctly to perform their functions. However, the mechanism of this localization is poorly understood. We report here the differential subnuclear localization of RNA strands of opposite polarity derived from the replicating Potato spindle tuber viroid (PSTVd). During replication, (+)- and (-)-strand viroid RNAs are produced. We found that in infected cultured cells and plants, the (-)-strand RNA was localized in the nucleoplasm, whereas the (+)-strand RNA was localized in the nucleolus as well as in the nucleoplasm with distinct spatial patterns. Furthermore, the presence of the (+)-PSTVd in the nucleolus caused the redistribution of a small nucleolar RNA. Our results support a model in which (1) the synthesis of the (-)- and (+)-strands of PSTVd RNAs occurs in the nucleoplasm, (2) the (-)-strand RNA is anchored in the nucleoplasm, and (3) the (+)-strand RNA is transported selectively into the nucleolus. Our results imply that the eukaryotic cell has a machinery that recognizes and localizes the opposite strands of an RNA, which may have broad ramifications in the RNA regulation of gene expression and the infection cycle of pathogenic RNAs and in the development of RNA-based methods to control gene expression as well as pathogen infection. PMID- 14555699 TI - Regulation of flowering time and floral organ identity by a MicroRNA and its APETALA2-like target genes. AB - MicroRNAs (miRNAs) are approximately 21-nucleotide noncoding RNAs that have been identified in both animals and plants. Although in animals there is direct evidence implicating particular miRNAs in the control of developmental timing, to date it is not known whether plant miRNAs also play a role in regulating temporal transitions. Through an activation-tagging approach, we demonstrate that miRNA 172 (miR172) causes early flowering and disrupts the specification of floral organ identity when overexpressed in Arabidopsis. miR172 normally is expressed in a temporal manner, consistent with its proposed role in flowering time control. The regulatory target of miR172 is a subfamily of APETALA2 (AP2) transcription factor genes. We present evidence that miR172 downregulates these target genes by a translational mechanism rather than by RNA cleavage. Gain-of-function and loss of-function analyses indicate that two of the AP2-like target genes normally act as floral repressors, supporting the notion that miR172 regulates flowering time by downregulating AP2-like target genes. PMID- 14555701 TI - Telomerase inhibition and cell growth arrest by G-quadruplex interactive agent in multiple myeloma. AB - OBJECTIVE: The aim of this study was to test the efficacy of telomerase inhibitor (TMPyP4 [tetra(N-methyl-4-pyridyl)-porphyrin chloride]; a G-quadruplex intercalating porphyrin) as a potential therapeutic agent for multiple myeloma. MATERIALS AND METHODS: We studied telomere length, telomerase activity, and effect of telomerase inhibition in multiple myeloma cells. Several myeloma cell lines were analyzed for telomerase activity, telomere length, and gene expression. Three myeloma cell lines (U266, ARH77, and ARD) were treated with TMPyP4 for 3-4 weeks. Viable cell number was assessed by trypan blue exclusion, and nature of cell death was determined by annexin labeling and/or DNA fragmentation. In situ oligo ligation technique was used to identify specific DNase I-type DNA cleavage. RESULTS: We report high telomerase activity and shortened telomeres in myeloma cells compared to normal B cells. We have also observed inhibition of telomerase activity, reduction in telomere length, and decline of myeloma cell growth, as measured by trypan blue dye exclusion, following exposure to TMPyP4. Exposure to porphyrin reduced telomerase activity of U266, ARH77, and ARD myeloma cells by 98%, 92%, and 99%, respectively. Exposure to porphyrin had no effect on viability for the first 14 days, followed by death of 75-90% of cells over the next 2 weeks. The nature of cell death was apoptotic, as determined by annexin and DNA nick labeling. Majority of cells showed DNA fragmentation specific to caspase-3-activated DNase I. CONCLUSIONS: These results demonstrate anti-proliferative activity of G-quadruplex intercalating agents, and suggest telomerase as an important therapeutic target for myeloma therapy. PMID- 14555702 TI - Differential effects of the proteasome inhibitor bortezomib on apoptosis and angiogenesis in human prostate tumor xenografts. AB - Bortezomib (Velcade, PS-341) is a dipeptide boronate inhibitor of the 26S proteasome developed for use in cancer therapy. Here we examined the effects of bortezomib on apoptosis and angiogenesis in derivatives of two popular human prostate cancer cell lines (LNCaP-Pro5 and PC3M-Pro4). Bortezomib strongly inhibited proliferation in both cell lines in vitro, but the PC3M-Pro4 cells were significantly more sensitive than the LNCaP-Pro5 cells to bortezomib-induced apoptosis. The compound also significantly inhibited the growth of LNCaP-Pro5 and LNCaP-Pro4 tumor xenografts, but the mechanisms involved in tumor growth inhibition differed in the two models. Bortezomib-treated LNCaP-Pro5 tumors displayed reduced microvessel densities and vascular endothelial cell growth factor secretion and high levels of endothelial cell apoptosis consistent with angiogenesis inhibition. In contrast, PC3M-Pro4 tumors were poorly vascularized at baseline, and bortezomib failed to induce significant changes in microvessel density, angiogenic factor secretion, or endothelial cell death in this model. Rather, growth inhibition in the PC3M-Pro4 tumors was associated with direct increases in tumor cell death. Together, our results confirm that bortezomib is active in preclinical models of human prostate cancer, but its effects on apoptosis versus angiogenesis are cell type dependent. PMID- 14555703 TI - Anti-angiogenic activity of human endostatin is HIF-1-independent in vitro and sensitive to timing of treatment in a human saphenous vein assay. AB - Endostatin is a 20-kDa endogenous angiogenesis inhibitor that has recently been shown to inhibit the expression of vascular endothelial growth factor (VEGF), an angiogenic growth factor that is up-regulated by hypoxia via the HIF-1 transcription factor complex. To determine if the anti-angiogenic activity of endostatin involves a modulation of the HIF-1/VEGF pathway in cancer cells, experiments were conducted to establish what effect endostatin has on HIF-1 activity, HIF-1alpha protein production, and cellular localization in prostate cancer cells and endothelial cells. Endothelial cell tube formation was inhibited by endostatin purchased from Calbiochem (San Diego, CA) but not endostatin obtained from EntreMed (Rockville, MD). Subsequent experiments using Calbiochem endostatin showed that it did not alter HIF-1alpha protein production or cellular localization in any of the cell lines tested, nor did it alter HIF-1 transactivational activity in hypoxia. Whether or not this is also true in vivo remains to be determined. Nevertheless, these data suggest that the anti angiogenic activity of endostatin is independent of the HIF-1/VEGF pathway. Immunocytochemical staining results do not indicate a decreased production of VEGF in Calbiochem endostatin-treated LNCaP or human umbilical vein endothelial cells (HUVEC). Treatment of rat aortic cross sections with human endostatin from Calbiochem resulted in a dose-dependent inhibition of microvessel outgrowth. Importantly, inhibition of vessel outgrowth by Calbiochem endostatin in a human saphenous vein angiogenesis assay required early treatment. In view of this in vitro data, we suggest that clinical trials involving endostatin treatment of late-stage disease may not adequately represent the efficacy of this drug in early-stage cancer. PMID- 14555704 TI - PG490-88, a derivative of triptolide, causes tumor regression and sensitizes tumors to chemotherapy. AB - Treatment of solid tumors with combinations of chemotherapeutic agents has not led to significant increases in long-term survival. Recent studies support a role for inhibitors of checkpoint arrest as a means to enhance the cytotoxicity of chemotherapy. We have shown previously that triptolide (PG490), an oxygenated diterpene derived from a Chinese medicinal plant, induces apoptosis in cultured tumor cells and sensitizes tumor cells to topoisomerase inhibitors by blocking p53-mediated induction of p21. Here we extend our studies to a tumor xenograft model and evaluate the efficacy and safety of PG490-88 (14-succinyl triptolide sodium salt), a water-soluble prodrug of PG490. We also look at the combination of PG490 or PG490-88 with CPT-11, a topoisomerase I inhibitor, in cultured cells and in the tumor xenograft model. We show that PG490-88 is a safe and potent antitumor agent when used alone, causing tumor regression of lung and colon tumor xenografts. We also show that PG490-88 acts in synergy with CPT-11 to cause tumor regression. A phase I trial of PG490-88 for solid tumors began recently and safety and optimal dosing data should accrue within the next 12 months. Our findings that PG490-88 causes tumor regression and that it acts in synergy with DNA-damaging chemotherapeutic agents suggest a role as an antineoplastic agent and chemosensitizer for the treatment of patients with solid tumors. PMID- 14555706 TI - MAC-321, a novel taxane with greater efficacy than paclitaxel and docetaxel in vitro and in vivo. AB - The taxanes, paclitaxel (PTX) and docetaxel (DTX), belong to a novel class of anticancer drugs that stabilize microtubules and lead to tumor cell death. While both agents are widely used for the treatment of lung, breast, and ovarian cancer, many tumor types are refractory or develop resistance to these drugs. We describe here a novel analogue of DTX, designated MAC-321 [Microtubule/Apoptosis/Cytotoxic: 5beta, 20-epoxy-1, 2alpha-, 4-, 7beta-, 10beta , 13alpha-hexahydroxytax-11-en-9-one 4 acetate 2 benzoate 7-propionate 13-ester with (2R,3S)-N-tertbutoxycarbonyl-3-(2-furyl)isoserine], that overcomes P glycoprotein-mediated resistance to PTX and DTX in preclinical model systems. Similar to PTX or DTX, MAC-321 enhanced the rate of tubulin polymerization in vitro and caused the bundling of microtubules in cells. MAC-321 inhibited proliferation of a panel of 14 tumor cell lines with minimal variation in potency (IC(50) = 2.2 +/- 1.4 nM; range = 0.6-5.3 nM). Unlike PTX or DTX, the IC(50) of MAC-321 did not vary in cells that expressed low to moderate levels of P glycoprotein. Even under extraordinary conditions in KB-V1 cells, which highly overexpress P-glycoprotein, resistance to MAC-321 was 80-fold compared with that of PTX (1400-fold) and DTX (670-fold). In addition, equivalent or less resistance to MAC-321 compared with PTX or DTX was observed in four cell lines that contain distinct point mutations within the taxane-binding site of beta-tubulin. Most importantly, MAC-321 displayed superior in vivo efficacy because: (a) MAC-321 either partially or completely inhibited tumor growth in three tumor models that overexpressed P-glycoprotein and were resistant to PTX; and (b) unlike PTX or DTX, MAC-321 was highly effective when given orally. MAC-321 was also highly effective when given as single i.v. dose. Our findings suggest that MAC-321, which is currently under clinical evaluation, may have broad therapeutic value. PMID- 14555705 TI - Kahalalide F, a new marine-derived compound, induces oncosis in human prostate and breast cancer cells. AB - Kahalalide F (KF) is a novel antitumor drug of marine origin under clinical investigation. KF showed a potent cytotoxic activity against a panel of human prostate and breast cancer cell lines, with IC(50) ranging from 0.07 micro M (PC3) to 0.28 micro M (DU145, LNCaP, SKBR-3, BT474, MCF7). Importantly, nontumor human cells (MCF10A, HUVEC, HMEC-1, IMR90) were 5-40 times less sensitive to the drug (IC(50) = 1.6-3.1 micro M). KF cytotoxicity did not correlate with the expression level of the multidrug resistance MDR1 and of the tyrosine kinase HER2/NEU, and only slightly by the anti-apoptotic BCL-2 protein. KF action was triggered rapidly by short pulse treatments (15 min caused 50% maximum cytotoxicity). Neither a general caspase inhibitor (Z-VAD-fmk) nor transcription or translation inhibitors (actinomycin D, cycloheximide) blocked KF action. Flow cytometry analysis revealed that KF induced neither cell-cycle arrest nor apoptotic hypodiploid peak. Using mitochondrial (JC-1)- and lysosomal (LysoTracker Green, Acridine Orange)-specific fluorophores, we detected loss of mitochondrial membrane potential and of lysosomal integrity following KF treatment. Confocal laser and electron microscopy revealed that KF-treated cells underwent a series of profound alterations including severe cytoplasmic swelling and vacuolization, dilation and vesiculation of the endoplasmic reticulum, mitochondrial damage, and plasma membrane rupture. In contrast, the cell nucleus showed irregular clumping of chromatin into small, condensed masses, while chromatin disappeared from other nuclear domains, but the nuclear envelope was preserved and no DNA degradation was detected. Together, these data indicate that KF induces cell death via oncosis preferentially in tumor cells. PMID- 14555707 TI - Sulindac metabolites induce caspase- and proteasome-dependent degradation of beta catenin protein in human colon cancer cells. AB - Colorectal cancer (CRC) is the second leading cause of cancer death in the USA. Accumulation of beta-catenin protein is nearly ubiquitous in colon adenomas and cancers, presumably due to mutations in the APC or beta-catenin genes that inhibit proteasome-dependent degradation of beta-catenin protein. Substantial clinical, epidemiological, and animal evidence indicate that sulindac and other non-steroidal anti-inflammatory drugs (NSAIDs) prevent the development of CRC. The mechanisms by which sulindac exerts its potent growth inhibitory effects against colon tumor cells are incompletely understood, but down-regulation of beta-catenin has been suggested as one potential mechanism. The goal of this study was to determine the mechanism of beta-catenin protein down-regulation by sulindac metabolites. Treatment of human colon cancer cell lines with apoptotic concentrations of sulindac metabolites (sulindac sulfide, sulindac sulfone) induced a dose- and time-dependent inhibition of beta-catenin protein expression. Inhibition of proteasome activity with MG-132 partially blocked the ability of sulindac sulfide and sulindac sulfone to inhibit beta-catenin protein expression. Pretreatment with the caspase inhibitor z-VAD-fmk blocked morphological signs of apoptosis as well as caspase cleavage, and also partially prevented beta-catenin degradation by sulindac metabolites. These effects occurred in cells with bi allelic APC mutation (SW480), with wild-type APC but mono-allelic beta-catenin mutation (HCT116) and in cells that lack expression of either COX-1 or -2 (HCT15). These results indicate that loss of beta-catenin protein induced by sulindac metabolites is COX independent and at least partially due to reactivation of beta-catenin proteasome degradation and partially a result of caspase activation during the process of apoptosis. PMID- 14555708 TI - Amifostine impairs p53-mediated apoptosis of human myeloid leukemia cells. AB - Amifostine is used as a cytoprotective agent in cancer treatments. Amifostine protects from apoptosis in some models and has been used as hematopoiesis stimulator in myeloid malignancies. As the apoptosis induced by many antitumoral agents is mediated by p53, we studied the effect of amifostine on p53-mediated apoptosis. We used human myeloid leukemia K562 and NB4 cells expressing the temperature-conditional p53-Val(135) mutant. Both cell lines undergo apoptosis at 32 degrees C due to the presence of p53 in wild-type conformation. We found that amifostine dramatically reduced apoptosis by p53 in both cell lines, as assessed by cell morphology, annexin V binding, fraction of sub-G(1) cells, and DNA laddering. To explore the mechanism responsible for this apoptosis protection, we tested the effect of amifostine on p53 transcriptional activity. We found that amifostine reduced p53-mediated transactivation of target promoters in NB4 and K562. Macroarray analysis confirmed that several p53 target genes as p21(Waf1), mdm2, gadd45, pig8, and pig3 were down-regulated at the mRNA level by amifostine in NB4 and K562. Also, c-myc was up-regulated by amifostine in K562 in the presence of p53, consistently with the impairment of p53-mediated apoptosis exerted by c-Myc in these cells. We conclude that amifostine impairs p53 dependent apoptosis of myeloid leukemia cells by reducing the activation of apoptosis-related genes. Our results open the possibility that amifostine could reduce the effectiveness of antitumoral treatments when it is dependent on active p53. PMID- 14555709 TI - Viral delivery of P450 reductase recapitulates the ability of constitutive overexpression of reductase enzymes to potentiate the activity of mitomycin C in human breast cancer xenografts. AB - Indolequinones such as mitomycin C (MMC) require enzymatic bioreduction to yield cytotoxic moieties. An attractive approach to overcome the potential variability in reductive bioactivation between tumors is to exploit specific enzyme bioreductive drug combinations in an enzyme-directed gene therapy (GDEPT) approach. To this end, human breast cancer cell lines (T47D, MDA468, and MDA231) that overexpress either DT-diaphorase (DTD) or NADPH:cytochrome P450 reductase (P450R) have been developed. Cytotoxicity of MMC was evaluated in the panel of cell lines following aerobic or anoxic exposure in vitro. DTD and/or P450R overexpression sensitized cells to MMC in air with no further increase in the cytotoxicity of MMC under anoxia. The most profound effect was seen in the MDA468 cells, where a 27-fold increase in potency was observed for MMC in the DTD overexpressing cell line. The MMC sensitization achieved through DTD and P450R overexpression in MDA468 cells was maintained in vivo. Xenografts established from the clonal lines exhibited significant tumor control following MMC treatment (treated/control [T/C] 17% and 51% for DTD and P450R xenografts, respectively) that was not seen in wild-type tumors (T/C 102%). Delivery of a clinically relevant adenoviral vector encoding P450R to MDA468 wild-type tumors yielded comparable P450R activity to that seen in the P450R clonal xenografts and resulted in greater MMC sensitization (T/C 46%). The model systems developed will facilitate the identification of novel indolequinone agents that are targeted toward a specific enzyme for bioactivation and are consequently of potential use in a GDEPT approach. PMID- 14555711 TI - Application of quantitative models from population biology and evolutionary game theory to tumor therapeutic strategies. AB - Quantitative models from population biology and evolutionary game theory frame the tumor-host interface as a dynamical microenvironment of competing tumor and normal populations. Through this approach, critical parameters that control the outcome of this competition are identified and the conditions necessary for formation of an invasive cancer are defined. Perturbations in these key parameters that destabilize the cancer solution of the state equations and produce tumor regression can be predicted. The mathematical models demonstrate significant theoretical limitations in therapies based solely on cytotoxic drugs. Because these approaches do not alter critical parameters controlling system dynamics, the tumor population growth term will remain positive as long as any individual cells are present so that the tumor will invariably recur unless all proliferative cells are killed. The models demonstrate that such total effectiveness is rendered unlikely by the genotypic heterogeneity of tumor populations (and, therefore, the variability of their response to such drugs) and the ability of tumor cells to adapt to these proliferation constraints by evolving resistant phenotypes. The mathematical models support therapeutic strategies that simultaneously alter several of the key parameters in the state equations. Furthermore, the models demonstrate that administration of cytotoxic therapies will, by reducing the tumor population density, create system dynamics more conducive to perturbations by biological modifiers. PMID- 14555710 TI - Transduction of green fluorescent protein increased oxidative stress and enhanced sensitivity to cytotoxic drugs in neuroblastoma cell lines. AB - Green fluorescent protein (GFP) is employed as a selection marker for gene transduction and to track tumor cells. Transduction of enhanced GFP (eGFP) into human neuroblastoma cell lines via a lentiviral vector significantly sensitized CHLA-20 (wild-type and functional TP53), and to a lesser extent CHLA-90 cells (multidrug-resistant, mutant, and nonfunctional TP53) to carboplatin, doxorubicin, etoposide, or melphalan, relative to cells transduced using the cell surface antigen CD80 as a selection marker. Total glutathione (GSH) was significantly up-regulated (1.8- to 2.8-fold) after eGFP (but not CD80) transduction in cell lines with, but not in those lacking, functional p53. Cytotoxicity of GSH depletion by buthionine sulfoximine in CHLA-20 (but not in CHLA-20-eGFP) was diminished by hypoxia (2% O(2)). Thus, oxidative stress produced by GFP selects for cells with up-regulated GSH in a p53-dependent manner, and also enhanced the cytotoxicity of anticancer drugs in neuroblastoma cell lines. Our data suggest caution when employing GFP-transduced cells to assess drug sensitivity and that using a cell surface antigen as a selection marker for gene transduction may perturb cells less than GFP. PMID- 14555712 TI - Preclinical models for defining efficacy of drug combinations: mapping the road to the clinic. PMID- 14555713 TI - Protein kinase A potentiates adrenal 4 binding protein/steroidogenic factor 1 transactivation by reintegrating the subcellular dynamic interactions of the nuclear receptor with its cofactors, general control nonderepressed 5/transformation/transcription domain-associated protein, and suppressor, dosage sensitive sex reversal-1: a laser confocal imaging study in living KGN cells. AB - The mechanism through which protein kinase A (PKA) potentiates the transactivation ability of adrenal 4 binding protein/steroidogenic factor 1 (Ad4BP/SF-1) is currently unclear. In the present study, we investigated the mechanism by applying laser confocal microscopy and fluorescence recovery after photobleaching technique. In KGN cells, forskolin (a PKA stimulator) could reorganize wild-type Ad4BP/SF-1, but not mutant Ad4BP/SF-1 (G35E), from a diffuse distribution pattern to foci formation in the nucleus. The subcellular distributions of GCN5 (general control nonderepressed) and TRRAP (transformation/transcription domain-associated protein), both of which were recently proved to be working in the same complex as the third class of nuclear receptor coactivators, were unexpectedly diffuse inside and outside the nucleus, respectively, when they were separately transfected. However TRRAP was translocated into the nucleus in the presence of GCN5, and together with GCN5 colocalized with Ad4BP/SF-1 in the same foci when PKA was activated. A luciferase assay also indicated that these two cofactors enhanced Ad4BP/SF-1 transactivation.Dosage-sensitive sex reversal (DAX-1) interacts with and thus inhibits Ad4BP/SF-1 transactivation. The coexistence of the two proteins dramatically altered their respective subnuclear distributions. They colocalized extensively, suggestive of binding, and Ad4BP/SF-1 was sharply immobilized when DAX-1 was coexpressed, whereas PKA could maintain mobility, as evidenced by Fluorescence Recovery After Photobleaching showing that Ad4BP/SF-1 mobility recovered after forskolin treatment.Therefore, the PKA signal pathway may modify the interaction between Ad4BP/SF-1 and its activators and repressor (GCN5 and TRRAP are integrated, whereas DAX-1 is disassociated), and thus stimulate the Ad4BP/SF-1 transactivation. PMID- 14555714 TI - Small conductance Ca2+-activated K+ channels formed by the expression of rat SK1 and SK2 genes in HEK 293 cells. AB - The rat SK1 gene (rSK1) does not form functional Ca2+-activated potassium channels when expressed alone in mammalian cell lines. Using a selective antibody to the rSK1 subunit and a yellow fluorescent protein (YFP) tag we have discovered that rSK1 expression produces protein that remains largely at intracellular locations. We tested the idea that rSK1 may need an expression partner, rSK2, in order to form functional channels. When rSK1 was co-expressed with rSK2 in HEK 293 cells it increased the current magnitude by 77 +/- 34% (as compared with cells expressing rSK2 alone). Co-expression of rSK1 with rSK2 also changed the channel pharmacology. The sensitivity of SK current to block by apamin was reduced approximately 16-fold from an IC50 of 94 pM (for SK2 alone) to 1.4 nM (for SK2 and SK1 together). The sensitivity to block by UCL 1848 (a potent small molecule blocker of SK channels) was similarly reduced, approximately 26-fold, from an IC50 of 110 pM to 2.9 nM. These data clearly demonstrate that rSK1 and rSK2 subunits interact. The most likely explanation for this is that the subunits are able to form heteromeric assemblies. PMID- 14555715 TI - Buffer kinetics shape the spatiotemporal patterns of IP3-evoked Ca2+ signals. AB - Ca2+ liberation through inositol 1,4,5-trisphosphate receptors (IP3Rs) plays a universal role in cell regulation, and specificity of cell signalling is achieved through the spatiotemporal patterning of Ca2+ signals. IP3Rs display Ca2+-induced Ca2+ release (CICR), but are grouped in clusters so that regenerative Ca2+ signals may remain localized to individual clusters, or propagate globally between clusters by successive cycles of Ca2+ diffusion and CICR. We used confocal microscopy and photoreleased IP3 in Xenopus oocytes to study how these properties are modulated by mobile cytosolic Ca2+ buffers. EGTA (a buffer with slow 'on-rate') speeded Ca2+ signals and 'balkanized' Ca2+ waves by dissociating them into local signals. In contrast, BAPTA (a fast buffer with similar affinity) slowed Ca2+ responses and promoted 'globalization' of spatially uniform Ca2+ signals. These actions are likely to arise through differential effects on Ca2+ feedback within and between IP3R clusters, because Ca2+ signals evoked by influx through voltage-gated channels were little affected. We propose that cell specific expression of Ca2+-binding proteins with distinct kinetics may shape the time course and spatial distribution of IP3-evoked Ca2+ signals for specific physiological roles. PMID- 14555716 TI - The modulation by 5-HT of glutamatergic inputs from the raphe pallidus to rat hypoglossal motoneurones, in vitro. AB - Decreases in the activity of 5-HT-containing caudal raphe neurones during sleep are thought to be partially responsible for the resultant disfacilitation of hypoglossal motoneurones. Whilst 5-HT has a direct excitatory action on hypoglossal motoneurones as a result of activation of 5-HT2 receptors, microinjection of 5-HT2 antagonists into the hypoglossal nucleus reduces motor activity to a much lesser extent compared to the suppression observed during sleep suggesting other transmitters co-localised in caudal raphe neurones may also be involved. The aim of the present study was therefore to characterise raphe pallidus inputs to hypoglossal motoneurones. Whole cell recordings were made from hypoglossal motoneurones in vitro. 5-HT evoked a direct membrane depolarisation (8.45 +/- 3.8 mV, P < 0.001) and increase in cell input resistance (53 +/- 40 %, P < 0.001) which was blocked by the 5-HT2 antagonist, ritanserin (2.40 +/- 2.7 vs. 7.04 +/- 4.6 mV). Stimulation within the raphe pallidus evoked a monosynaptic EPSC that was significantly reduced by the AMPA/kainate antagonist, NBQX (22.8 +/- 16 % of control, P < 0.001). In contrast, the 5-HT2 antagonist, ritanserin, had no effect on the amplitude of these EPSCs (106 +/- 31 % of control, P = n.s.). 5-HT reduced these EPSCs to 50.0 +/- 13 % of control (P < 0.001), as did the 5-HT1A agonist, 8-OH-DPAT (52.5 +/- 17 %, P < 0.001) and the 5-HT1B agonist, CP 93129 (40.6 +/- 29 %, P < 0.01). 8-OH-DPAT and CP 93129 increased the paired pulse ratio (1.38 +/- 0.27 to 1.91 +/- 0.54, P < 0.05 & 1.27 +/- 0.08 to 1.44 +/- 0.13, P < 0.01 respectively) but had no effect on the postsynaptic glutamate response (99 +/- 4.4 % and 100 +/- 2.5 %, P = n.s.). They also increased the frequency (P < 0.001), but not the amplitude, of miniature glutamatergic EPSCs in hypoglossal motoneurones. These data demonstrate that raphe pallidus inputs to hypoglossal motoneurones are predominantly glutamatergic in nature, with 5-HT decreasing the release of glutamate from these projections as a result of activation of 5-HT1A and/or 5-HT1B receptors located on presynaptic terminals. PMID- 14555717 TI - A juvenile form of postsynaptic hippocampal long-term potentiation in mice deficient for the AMPA receptor subunit GluR-A. AB - In adult mice, long-term potentiation (LTP) of synaptic transmission at CA3-to CA1 synapses induced by tetanic stimulation requires L-alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionate (AMPA) receptors containing GluR-A subunits. Here, we report a GluR-A-independent form of LTP, which is comparable in size to LTP in wild-type mice at postnatal day 14 (P14) but diminishes between P14 and P42 in brain slices of GluR-A-deficient mice. The GluR-A-independent form of LTP is sensitive to D(-)-2-amino-5-phosphonopentanoic acid (D-AP5), but lacks short-term potentiation (STP) and can also be observed in the pairing induction protocol. As judged by unaltered paired-pulse facilitation, this LTP form is postsynaptically expressed despite depleted extrasynaptic AMPA receptor pools with reduced levels of GluR-B, which accumulates in somata and synapses of CA1 pyramidal neurons in GluR-A-deficient mice. Our results show that in the developing hippocampus synaptic plasticity can be expressed by AMPA receptors lacking the GluR-A subunit. PMID- 14555718 TI - Nicotine is highly effective at producing desensitization of rat alpha4beta2 neuronal nicotinic receptors. AB - We examined desensitization by acetylcholine (ACh) and nicotine at the rat alpha4beta2 neuronal nicotinic receptor stably expressed in HEK cells. For both agonists, the decay in response due to desensitization ('onset') was best fitted by the sum of two exponentials with the fast component dominant at concentrations > 1 microM. The time constants for onset were similar for both agonists, and showed little concentration dependence over the range of 0.1-100 microM. Recovery from desensitization also showed two exponential components. In contrast to the similarity in onset, nicotine produced longer lasting desensitization, resulting from an increase in the proportion of receptors in the slowly recovering population and from an increase in the time constant for the slow recovery process. The proportion of receptors in the slowly recovering population increased as the duration of the desensitizing pulse increased. Desensitization was also induced by low concentrations of agonist, with no apparent macroscopic response. A 100 s application of 10 nM nicotine desensitized 70 % of the peak response, while 100 s of 10 nM ACh desensitized only 15 %. At higher concentrations of agonist, which result in a macroscopic response, desensitization in the absence of activation also can occur. Nicotine is a very potent and efficacious desensitizing agent at this neuronal nicotinic receptor. PMID- 14555720 TI - Long vasodilator reflexes projecting through the myenteric plexus in guinea-pig ileum. AB - This study examined enteric neural reflexes activating submucosal cholinergic vasodilator motoneurons, which innervate the final resistance vessels regulating mucosal blood flow. Videomicroscopy was employed to monitor dilatation of submucosal arterioles in in vitro preparations from guinea-pig ileum. Balloon distension of intact lumen evoked reflex vasodilatation and flat sheet preparations were employed to separate mucosal mechanical stimulation from intestinal distension. Mucosal stroking and balloon distension of the orad segment evoked vasodilatations > 1.5 cm from the stimulating site. Mucosal stimulation was blocked by combined 5-HT3/5HT4 antagonists but distension-evoked responses were unaffected. Distension-evoked responses were also unaffected by nifedipine (5 microM) or nifedipine (1 microM) and wortmannin (300 nM), suggesting stretch activation rather than stretch-activated contraction was involved. Mucosal and distension-evoked responses were completely blocked when the myenteric plexus was surgically lesioned and were significantly inhibited by hexamethonium. The muscarinic antagonist 4-DAMP, which inhibits vasodilatations evoked by submucosal cholinergic vasodilator neurons, blocked dilatations elicited by mucosal stimulation and balloon distension. Maximal dilatations evoked with either sensory modality could be further enhanced when stimulated with the second modality. Dilatations evoked by stimulation of the aborad segment were similar to those elicited in the orad segment. In conclusion, sensory mechanisms in the mucosa and muscularis propria activate vasodilator pathways in the myenteric plexus which project for significant distances in both ascending and descending directions before innervating submucosal arterioles. These reflexes could co-ordinate mucosal blood flow during multiple motor events such as peristalsis and intestinal mixing between propulsive events. PMID- 14555721 TI - Activation of M1 muscarinic receptors triggers transmitter release from rat sympathetic neurons through an inhibition of M-type K+ channels. AB - Acetylcholine has long been known to excite sympathetic neurons via M1 muscarinic receptors through an inhibition of M-currents. Nevertheless, it remained controversial whether activation of muscarinic receptors is also sufficient to trigger noradrenaline release from sympathetic neurons. In primary cultures of rat superior cervical ganglia, the muscarinic agonist oxotremorine M inhibited M currents with half-maximal effects at 1 microM and induced the release of previously incorporated [3H]noradrenaline with half-maximal effects at 10 microM. This latter action was not affected by the nicotinic antagonist mecamylamine which, however, abolished currents through nicotinic receptors elicited by high oxotremorine M concentrations. Ablation of the signalling cascades linked to inhibitory G proteins by pertussis toxin potentiated the release stimulating effect of oxotremorine M, and the half-maximal concentration required to stimulate noradrenaline release was decreased to 3 microM. Pirenzepine antagonized the inhibition of M-currents and the induction of release by oxotremorine M with identical apparent affinity, and both effects were abolished by the muscarinic toxin 7. These results indicate that one muscarinic receptor subtype, namely M1, mediates these two effects. Retigabine, which enhances M currents, abolished the release induced by oxotremorine M, but left electrically induced release unaltered. Moreover, retigabine shifted the voltage-dependent activation of M-currents by about 20 mV to more negative potentials and caused 20 mV hyperpolarisations of the membrane potential. In the absence of retigabine, oxotremorine M depolarised the neurons and elicited action potential discharges in 8 of 23 neurons; in its presence, oxotremorine M still caused equal depolarisations, but always failed to trigger action potentials. Action potential waveforms caused by current injection were not affected by retigabine. These results indicate that the inhibition of M-currents is the basis for the stimulation of transmitter release from sympathetic neurons via M1 muscarinic receptors. PMID- 14555722 TI - Acute stress modulates the histamine content of mast cells in the gastrointestinal tract through interleukin-1 and corticotropin-releasing factor release in rats. AB - Stress results in activation of the hypothalamic pituitary adrenal axis and affects illnesses such as neuroinflammatory syndrome. In vivo acute stress (restraint stress) induces gastrointestinal function disturbances through colonic mast cell activation. This study investigated the effect of acute stress in histamine content of colonic mast cells, and the central role of interleukin-1 (IL-1) and corticotropin-releasing factor (CRF) in this effect. After a restraint stress session colonic segments were isolated and submitted to three protocols: (i) determination of histamine levels by radioimmunoassay (RIA) after incubation with 48/80 compound, (ii) evaluation by histology of mucosal mast cell (MMC) number and (iii) determination of histamine immunoreactivity of MMC. These procedures were conducted (1) in sham or stressed rats, (2) in stressed rats previously treated with intracerebroventricular (I.C.V.) IL-1ra or alpha-helical CRF9-41, (3) in naive rats pretreated with I.C.V. rhIL-1beta or CRF and (4) in rats treated with central IL-1beta and CRF plus alpha-helical CRF and IL-1ra, respectively (cross-antagonism reaction). Acute stress increases histamine content in colonic mast cells, without degranulation. I.C.V. pretreatment with IL 1ra or alpha-helical CRF9-41 blocked stress-induced mast cell histamine content increase. Both I.C.V. rhIL-1beta and CRF injections reproduced the stress-linked changes. I.C.V. treatment with CRF antagonist blocked I.C.V. rhIL-1beta-induced mast cell histamine content increase, whereas central IL-1ra did not affect stress events induced by I.C.V. CRF administration. These results suggest that in rats acute stress increases colonic mast cell histamine content. This effect is mediated by the release in cascade in the brain first of IL-1 and secondly of CRF. PMID- 14555723 TI - Null mutation of gp91phox reduces muscle membrane lysis during muscle inflammation in mice. AB - Muscle inflammation is a common feature in muscle injury and disease. Recently, investigators have speculated that inflammatory cells may increase or decrease muscle damage following modified muscle use, although there are few experimental observations to confirm either possibility. In the present study, a null mutation of gp91phox in neutrophils prevented superoxide production in cytotoxicity assays in which muscle cells were targets, and prevented most neutrophil-mediated cytolysis of muscle cells in comparison to wild-type neutrophils in vitro. We further tested whether deficiency in superoxide production caused a decrease in muscle membrane damage in vivo during modified muscle use. Gp91phox null mutant mice and wild-type mice were subjected to 10 days of muscle hindlimb unloading followed by reloading through return to normal locomotion, which induced muscle membrane lesions and muscle inflammation. Membrane lesions were quantified by measuring the presence of extracellular marker dye in reloaded soleus muscle fibres. There was a 90 % reduction in the number of fibres showing extensive membrane injury in gp91phox null mice compared to controls. Mutation of gp91phox did not change the concentration of neutrophils or macrophages in the reloaded muscle. Furthermore, muscle fibre growth during the reloading period was unaffected by the reduction in membrane injury. Together, these findings show that neutrophils can induce muscle membrane lysis through superoxide-mediated events, and indicate that superoxide-mediated membrane damage in vivo is not required for myeloid cell chemotaxis or muscle growth during muscle reloading. PMID- 14555724 TI - Small- and intermediate-conductance calcium-activated K+ channels provide different facets of endothelium-dependent hyperpolarization in rat mesenteric artery. AB - Activation of both small-conductance (SKCa) and intermediate-conductance (IKCa) Ca2+-activated K+ channels in endothelial cells leads to vascular smooth muscle hyperpolarization and relaxation in rat mesenteric arteries. The contribution that each endothelial K+ channel type makes to the smooth muscle hyperpolarization is unknown. In the presence of a nitric oxide (NO) synthase inhibitor, ACh evoked endothelium and concentration-dependent smooth muscle hyperpolarization, increasing the resting potential (approx. -53 mV) by around 20 mV at 3 microM. Similar hyperpolarization was evoked with cyclopiazonic acid (10 microM, an inhibitor of sarcoplasmic endoplasmic reticulum calcium ATPase (SERCA)) while 1-EBIO (300 microM, an IKCa activator) only increased the potential by a few millivolts. Hyperpolarization in response to either ACh or CPA was abolished with apamin (50 nM, an SKCa blocker) but was unaltered by 1-[(2 chlorophenyl) diphenylmethyl]-1H-pyrazole (1 microM TRAM-34, an IKCa blocker). During depolarization and contraction in response to phenylephrine (PE), ACh still increased the membrane potential to around -70 mV, but with apamin present the membrane potential only increased just beyond the original resting potential (circa -58 mV). TRAM-34 alone did not affect hyperpolarization to ACh but, in combination with apamin, ACh-evoked hyperpolarization was completely abolished. These data suggest that true endothelium-dependent hyperpolarization of smooth muscle cells in response to ACh is attributable to SKCa channels, whereas IKCa channels play an important role during the ACh-mediated repolarization phase only observed following depolarization. PMID- 14555726 TI - Calcium mobilisation and CCK secretion induced by modified fatty acids and latex microspheres reveal dual receptor mechanisms for lipid stimulation of STC-1 cells. AB - How fatty acids stimulate enteroendocrine cells to release cholecystokinin (CCK) is largely unknown. Recently, we proposed that the murine enteroendocrine cell line, STC-1, responds to insoluble fatty acid aggregates rather than fatty acid monomers in solution. This hypothesis led to two testable predictions. First, other insoluble particles of similar size but unrelated to fatty acid may be able to stimulate STC-1 cells in a similar fashion to dodecanoic acid and second, fatty acid sensing in STC-1 cells should be fairly insensitive to chemical modifications of the fatty acid as long as these modifications do not greatly alter the ability of the molecule to form insoluble aggregates. We used several analogues of dodecanoic acid and several varieties of latex microsphere (varying in size and surface charge) to see whether the predictions of our model hold. We found that while there was at least one latex microsphere that could induce CCK secretion and calcium mobilisation in STC-1 cells, there was a very poor correlation between the presence of insoluble aggregates and a cellular response. Instead the most important property, determining the potency of fatty acid analogues as stimulants of CCK secretion, was their amphipathicity. Removal of either the polar head or lipophilic tail completely abolished the ability of a given fatty acid analogue to stimulate STC-1 cells. These data suggested that while fatty acids can stimulate cells as aggregates, they may also be acting in monomeric form with the oil:water partitioning coefficient playing a crucial role. We finally resolved this issue with the observation that the sulfate ion greatly altered the response of STC-1 cells to monomeric dodecanoic acid. In the presence of sulfate, STC-1 cells will only respond to dodecanoic acid aggregates whereas when sulfate is replaced with chloride the cells clearly respond to dodecanoic acid monomers which are completely in solution. In summary, we propose that dodecanoic acid can stimulate STC-1 cells via two separate pathways one involving fatty acid monomers in solution and one involving fatty acid aggregates. Which pathway dominates depends on the presence of sulfate in the extracellular medium. PMID- 14555725 TI - The mechanism of the force response to stretch in human skinned muscle fibres with different myosin isoforms. AB - Force enhancement during lengthening of an active muscle, a condition that normally occurs during locomotion in vivo, is attributed to recruitment of myosin heads that exhibit fast attachment to and detachment from actin in a cycle that does not imply ATP splitting. We investigated the kinetic and mechanical features of this cycle in Ca(2+) activated single skinned fibres from human skeletal muscles containing different myosin heavy chain (MHC) isoforms, identified with single-fibre gel electrophoresis. Fibres were activated by using a new set-up that allows development of most of the tension following a temperature jump from 0-1 degrees C to the test temperature (approximately 12 degrees C). In this way we could prevent the development of sarcomere non-uniformity and record sarcomere length changes with a striation follower in any phase of the mechanical protocol. We found that: (i) fibres with fast MHC isoforms develop 40-70% larger isometric forces than those with slow isoforms, as a result of both a larger fraction of force-generating myosin heads and a higher force per head; (ii) in both slow and fast fibres, force enhancement by stretch is due to recruitment of myosin head attachments, without increase in strain per head above the value generated by the isometric heads; and (iii) the extent of recruitment is larger in slow fibres than in fast fibres, so that the steady force and power output elicited by lengthening become similar, indicating that mechanical and kinetic properties of the actin-myosin interactions under stretch become independent of the MHC isoform. PMID- 14555727 TI - Central command is capable of modulating sweating from non-glabrous human skin. AB - Isometric handgrip exercise (IHG) increases sweating rate without changing core or skin temperatures. The contribution of central command resulting in increases in sweating rate during IHG is unknown. To investigate this question, seven subjects performed IHG (35 % maximum voluntary contraction (MVC) for 2 min) followed by 2-min of post-exercise ischaemia (PEI), with and without partial neuromuscular blockade (PNB). PNB was performed to augment central command during the IHG bout. These trials were conducted while the subject was normothermic, mildly heated, and moderately heated. On the non-exercising arm, forearm sweating rate was monitored over a microdialysis membrane perfused with neostigmine (acetylcholinesterase inhibitor), and at an adjacent untreated site. In normothermia with PNB, despite reduced force production during IHG (17 +/- 9 versus 157 +/- 13 N; P < 0.001), the elevation in sweating rate at the neostigmine-treated site was greater relative to the control IHG bout (P < 0.05). During subsequent PEI, for the PNB trial mean arterial blood pressure (MAP) and sweating rate returned towards pre-IHG levels, while during the control trial these variables remained elevated. During IHG while mildly heated, the elevation in sweating rate was greater during the PNB trial relative to the control trial. In contrast, during moderate heating sweating increased during IHG for both trials, however the elevation in sweating rate during the PNB trial was not greater than during the control trial. These results suggest that central command is capable of modulating sweating rate in all thermal conditions, however its effect is reduced when body temperatures and/or sweating rate are substantially elevated. PMID- 14555728 TI - Shaping the excitability of human motor cortex with premotor rTMS. AB - Recent studies have shown that low-frequency repetitive transcranial magnetic stimulation (rTMS) to the left dorsal premotor cortex has a lasting influence on the excitability of specific neuronal subpopulations in the ipsilateral primary motor hand area (M1(HAND)). Here we asked how these premotor to motor interactions are shaped by the intensity and frequency of rTMS and the orientation of the stimulating coil. We confirmed that premotor rTMS at 1 Hz and an intensity of 90% active motor threshold (AMT) produced a lasting decrease in corticospinal excitability probed with single-pulse TMS over the left M1(HAND). Reducing the intensity to 80% AMT increased paired-pulse excitability at an interstimulus interval (ISI) of 7 ms. Opposite effects occurred if rTMS was given at 5 Hz: at 90% AMT, corticospinal excitability increased; at 80% AMT, paired pulse excitability at ISI = 7 ms decreased. No effects were seen if rTMS was applied at the same intensities to prefrontal or primary motor cortices. These findings indicate that the intensity of premotor rTMS determines the net effect of conditioning on distinct populations of neurones in the ipsilateral M1(HAND), but it is the frequency of rTMS that determines the direction of the induced change. By selecting the appropriate intensity and frequency, premotor rTMS allows to induce a predictable up- or down-regulation of the excitability in distinct neuronal circuits of human M1(HAND). PMID- 14555731 TI - Energy budget of hepatocytes from Antarctic fish (Pachycara brachycephalum and Lepidonotothen kempi) as a function of ambient CO2: pH-dependent limitations of cellular protein biosynthesis? AB - Scenarios of rising CO2 concentration in surface waters due to atmospheric accumulation of anthropogenic CO2, or in the deep sea due to anticipated industrial dumping of CO2, suggest that hypercapnia (elevated partial pressure of CO2) will become a general stress factor in aquatic environments, with largely unknown effects on species survival and well being, especially in cold and deep waters. For an analysis of CO2 effects at the cellular level, isolated hepatocytes were prepared from two representatives of the Antarctic fish fauna, Pachycara brachycephalum and Lepidonotothen kempi. Correlated changes in energy and protein metabolism were investigated by determining the rates of oxygen consumption at various levels of PCO2, of intra- and extracellular pH, and after inhibition of protein synthesis by cycloheximide. A decrease in extracellular pH (pHe) from control levels (pHe 7.90) to pHe 6.50 caused a reduction in aerobic metabolic rate of 34-37% under both normocapnic and hypercapnic conditions. Concomitantly, protein biosynthesis was inhibited by about 80% under conditions of severe acidosis in hepatocytes from both species. A parallel drop in intracellular pH probably mediates this effect. In conclusion, the present data indicate that elevated PCO2 may limit the functional integrity of the liver due to a pronounced depression in protein anabolism. This process may contribute to the limits of whole-animal tolerance to raised CO2 levels. PMID- 14555732 TI - van der Waals and hygroscopic forces of adhesion generated by spider capture threads. AB - Cribellar thread is the most primitive type of sticky prey capture thread found in aerial spider webs. Its outer surface is formed of thousands of fine fibrils that issue from a cribellum spinning field. The fibrils of primitive cribellar thread are cylindrical, whereas those of derived threads have nodes. Cribellar threads snag on insect setae but also adhere to smooth surfaces. A previous study showed empirically that cylindrical fibrils use only van der Waals forces to stick to smooth surfaces, as their stickiness is the same under different humidity. By contrast, noded fibrils are stickier under high humidity, where they are presumed to adsorb atmospheric water and implement hygroscopic (capillary) adhesion. Here, we model thread stickiness according to these two adhesive mechanisms. These models equate stickiness with the force necessary to overcome the adhesion of fibril contact points in a narrow band along each edge of the contact surface and to initiate peeling of the thread from the surface. Modeled and measured thread stickiness values are similar, supporting the operation of the hypothesized adhesive forces and portraying an important transition in the evolution of spider threads. Cribellar threads initially relied only on van der Waals forces to stick to smooth surfaces. The appearance of fibril nodes introduced hydrophilic sites that implemented hygroscopic force and increased thread stickiness under intermediate and high humidity. PMID- 14555729 TI - The central nucleus of the amygdala modulates gut-related neurons in the dorsal vagal complex in rats. AB - Using retrograde tract-tracing and electrophysiological methods, we characterized the anatomical and functional relationship between the central nucleus of the amygdala and the dorsal vagal complex. Retrograde tract-tracing techniques revealed that the central nucleus of the amygdala projects to the dorsal vagal complex with a topographic distribution. Following injection of retrograde tracer into the vagal complex, retrogradely labelled neurons in the central nucleus of the amygdala were clustered in the central portion at the rostral level and in the medial part at the middle level of the nucleus. Few labelled neurons were seen at the caudal level. Electrical stimulation of the central nucleus of the amygdala altered the basal firing rates of 65 % of gut-related neurons in the nucleus of the solitary tract and in the dorsal motor nucleus of the vagus. Eighty-one percent of the neurons in the nucleus of the solitary tract and 47 % of the neurons in the dorsal motor nucleus were inhibited. Electrical stimulation of the central nucleus of the amygdala also modulated the response of neurons in the dorsal vagal complex to gastrointestinal stimuli. The predominant effect on the neurons of the nucleus of the solitary tract was inhibition. These results suggest that the central nucleus of the amygdala influences gut-related neurons in the dorsal vagal complex and provides a neuronal circuitry that explains the regulation of gastrointestinal activity by the amygdala. PMID- 14555733 TI - Sensory innervation of the ovotestis in the snail Helix aspersa. AB - Because oviposition in the land snail Helix aspersa is a metabolically expensive process coupled to a high fixed cost, one expects oviposition to occur only when the clutch size surpasses a minimum value at which the reproductive benefit exceeds the cost. We propose that neural innervation of the gonad allows H. aspersa to monitor oocyte production and ensure an adequate supply of gametes prior to ovulation. The ovotestis is innervated by a branch of the intestinal nerve in which the majority of axon fibres measure <0.2 microm in diameter. We found a strong positive correlation between the number of mature oocytes in the ovotestis and the frequency of spontaneous afferent spikes in the nerve branch. Tactile stimulation of the ovotestis resulted in a 20-fold increase in afferent spikes and an efferent reflex directed towards the ovotestis and the pericardium. Afferent activity also increased 10-fold after an experimentally induced increase in the volume of the ovotestis. These results suggest that the growing oocytes expand the walls of the acini and trigger action potentials in the mechanosensitive nerve terminals that lie within the acinar walls. We hypothesize that the resulting tonic signal is permissive for ovulation. In addition, a phasic sensory signal may occur during ovulation to trigger CNS motor output related to oviposition. PMID- 14555735 TI - Burrow surveillance in fiddler crabs. I. Description of behaviour. AB - When defending resources, animals need to reliably detect and identify potential competitors. Animals that live at high population densities would be expected to be efficient in this aspect of resource defence since the time lost in false alarms could be substantial and the failure of identifying a competitor could be very costly. How does an animal decide whether another animal is or is not a threat to a resource or a territory? Fiddler crabs [Uca vomeris (McNeill)] operate from burrows that they guard and defend vigorously against other crabs. The crabs live in dense populations, with many animals inhabiting one square metre of mudflat. We describe here the behavioural responses of foraging crabs to repeated presentations of small crab-like dummies approaching their burrows. We explore the relationship between the probability and the timing of burrow defence responses, the crab's behavioural state, and the visual appearance and direction of approach of the dummies. We find that the probability of response of resident crabs is independent of the relative position of crab and dummy but is strongly affected by the dummy's position and movement direction relative to the crab's burrow. The critical stimuli are the dummy's distance from the crab's burrow and whether the dummy is moving towards the burrow or not. The response distance (dummy-burrow distance) increases with the crab's own distance from the burrow, indicating that the crabs modify their assessment of threat depending on their own distance away from the burrow. Differences in dummy size and brightness do not affect the probability or the timing of the response. We discuss these results in the context of fiddler crab social life and, in a companion paper, identify the visual and non-visual cues involved in burrow defence. PMID- 14555734 TI - Effects of loading and size on maximum power output and gait characteristics in geckos. AB - Stride length, stride frequency and power output are all factors influencing locomotor performance. Here, we first test whether mass-specific power output limits climbing performance in two species of geckos (Hemidactylus garnoti and Gekko gecko) by adding external loads to their bodies. We then test whether body size has a negative effect on mass-specific power output. Finally, we test whether loading affects kinematics in both gecko species. Lizards were induced to run vertically on a smooth wooden surface with loads of 0-200% of body mass (BM) in H. garnoti and 0-100% BM in G. gecko. For each stride, we calculated angular and linear kinematics (e.g. trunk angle, stride length), performance (maximum speed) and mean mass-specific power output per stride. The addition of increasingly large loads caused an initial increase in maximum mass-specific power output in both species, but for H. garnoti, mass-specific power output remained constant at higher loads (150% and 200% BM), even though maximum velocity declined. This result, in combination with the fact that stride frequency showed no evidence of leveling off as speed increased in either species, suggests that power limits maximum speed. In addition, the large gecko (G. gecko) produced significantly less power than the smaller H. garnoti, despite the fact that both species ran at similar speeds. This difference disappeared, however, when we recalculated power output based on higher maximum speeds for unloaded G. gecko moving vertically obtained by other researchers. Finally, the addition of external loads did not affect speed modulation in either species: both G. gecko and H. garnoti increase speed primarily by increasing stride frequency, regardless of loading condition. For a given speed, both species take shorter but more strides with heavier loads, but for a given load, G. gecko attains similar speeds to H. garnoti by taking longer but fewer strides. PMID- 14555736 TI - Burrow surveillance in fiddler crabs. II. The sensory cues. AB - Using crab-like dummies, we have shown previously that fiddler crabs [Uca vomeris (McNeill)] defend their burrows against intruders in a burrow-centred frame of reference. The crabs respond whenever an intruder approaches to within a certain distance of the burrow entrance, and this distance is independent of the approach direction. We show here that the crabs combine information from the path integration system on the location of their invisible burrow and visual information on the retinal position of an intruder to make this allocentric judgement. Excluding all alternative visual cues, we propose that the crabs employ a small set of matched visual filters to determine the relationship between a crab-like object and the invisible burrow. To account for the constantly varying distance between the crabs and their burrows, the state of the path integrator may select the appropriate one of these retinal 'warning zones'. We have shown before that burrow-owning fiddler crabs are extremely responsive to potential burrow snatchers, which we simulated with crab-like dummies moving across the substratum towards the burrow of residents. The crab's decision to respond to these dummies depends mainly on the spatial arrangement between itself, its burrow and the approaching dummy. The most important factor predicting response probability is the dummy's distance from the crab's burrow: the crabs are more likely to respond the closer the dummy approaches the burrow. The dummy-burrow distance not only determines the overall response probability but also the timing of burrow defence responses (i.e. when the crabs decide to react). Most interestingly, this response distance is independent of the dummy's direction of approach to the burrow. In addition, the crabs respond earlier to a dummy approaching their burrow if they themselves are further away from it, indicating that knowledge of their own distance from the burrow has an influence on their decision to respond. These results raise a number of interesting issues, which are the focus of this paper, regarding the cues and the information used by the crabs in burrow surveillance. PMID- 14555737 TI - Neural images of pursuit targets in the photoreceptor arrays of male and female houseflies Musca domestica. AB - Male houseflies use a sex-specific frontal eye region, the lovespot, to detect and pursue mates. We recorded the electrical responses of photoreceptors to optical stimuli that simulate the signals received by a male or female photoreceptor as a conspecific passes through its field of view. We analysed the ability of male and female frontal photoreceptors to code conspecifics over the range of speeds and distances encountered during pursuit, and reconstructed the neural images of these targets in photoreceptor arrays. A male's lovespot photoreceptor detects a conspecific at twice the distance of a female photoreceptor, largely through better optics. This detection distance greatly exceeds those reported in previous behavioural studies. Lovespot photoreceptors respond more strongly than female photoreceptors to targets tracked during pursuit, with amplitudes reaching 25 mV. The male photoreceptor also has a faster response, exhibits a unique preference for stimuli of 20-30 ms duration that selects for conspecifics and deblurs moving images with response transients. White-noise analysis substantially underestimates these improvements. We conclude that in the lovespot, both optics and phototransduction are specialised to enhance and deblur the neural images of moving targets, and propose that analogous mechanisms may sharpen the neural image still further as it is transferred to visual interneurones. PMID- 14555738 TI - Chemosensory tuning to a host recognition cue in the facultative specialist larvae of the moth Manduca sexta. AB - Larvae of Manduca sexta are facultative specialists on plants in the family Solanaceae. Larvae reared on solanaceous foliage develop a strong preference for their host; otherwise, they remain polyphagous. The host-specific recognition cue in potato foliage for Manduca larvae is the steroidal glycoside, indioside D. Two pairs of galeal taste sensilla, the lateral and medial sensilla styloconica, are both necessary and sufficient for the feeding preferences of host-restricted larvae. We conducted electrophysiological tip recordings from sensilla of solanaceous or wheat germ diet-reared larvae. For each animal, recordings of the responses to indioside D, glucose, tomatine and KCl were compared. All responses included both phasic and tonic portions. The sensilla styloconica of solanaceous reared larvae were tuned to indioside D, defined as maintaining a high sensitivity to indioside D, while showing lower sensitivity to other plant compounds. Half of the sensillar neurons of solanaceous-reared larvae were 'tuned' to indioside D, whereas those of wheat germ diet-reared larvae were not. The different responses between the two types of animals were a result of changes of individual receptor cells' responses in the sensilla. Feeding on solanaceous foliage therefore appears to result in a modification of the physiological responses of individual taste receptor cells that causes them to be tuned to the host-recognition cue indioside D. We propose that this tuning is the basis for the host-restricted larvae's strong behavioral preferences for solanaceous foliage. PMID- 14555739 TI - Mechanosensory-induced behavioural gregarization in the desert locust Schistocerca gregaria. AB - Desert locusts show an extreme form of phenotypic plasticity, changing between a cryptic solitarious phase and a swarming gregarious phase that differ in many aspects of behaviour, physiology and appearance. Solitarious locusts show rapid behavioural phase change in response to tactile stimulation directed to the hind femora. Repeatedly touching as little as one quarter of the anterior (outer) surface area of a hind femur produced full behavioural gregarization within 4 h. Solitarious locusts have approximately 30% more mechanosensory trichoid sensilla on the hind femora than do gregarious locusts but have similar or fewer numbers of sensilla elsewhere on the legs. Tactile stimulation of a hind femur in solitarious locusts that had been restrained so that they could not move their legs failed to induce any behavioural gregarization. Patterned electrical stimulation of metathoracic nerve 5, which innervates the hind leg, however, produced full gregarization in restrained locusts. Our data show for the first time that the gregarizing signal combines both exteroceptive and proprioceptive components, which travel in both nerves 5B1 and 5B2, and provides us with a powerful experimental method with which to elicit and study neuronal plasticity in this system. Acetic acid odour, a strong chemosensory stimulus that activates the same local processing pathways as exteroceptive stimuli, failed to elicit behavioural gregarization, suggesting an early segregation in the central nervous system of the mechanosensory signals that leads to gregarization. PMID- 14555740 TI - Effect of coronary perfusion on the basal performance, volume loading and oxygen consumption in the isolated resistance-headed heart of the trout Oncorhynchus mykiss. AB - Basal performance, volume loading response and oxygen consumption were determined in a resistance-headed preparation of the isolated trout heart. Two groups of hearts were used: the +CF group, in which the coronary vascular tree was perfused with a flow directly related to the pressure generated by the heart, and the -CF group, in which the coronary flow was set to zero. As a criterion for setting basal performance, the atrial input pressure was set in order to induce the ventricle to produce a cardiac output of 15 ml min(-1) kg(-1). Once basal conditions were obtained, the preparation was perfused for 30 min, and atrial and aortic pressure, cardiac output, heart rate, coronary pressure and coronary flow were determined at 5 min intervals. At the onset of perfusion, there was no difference in the basal performance between the two groups: the same preload was necessary to get the same cardiac output in both perfusion groups. None of the other performance parameters determined were different. However, after only 5 min of perfusion, the -CF hearts displayed significant adjustments, with increased atrial preload and ventricular preload (mean atrial pressure), and a significant decrease in cardiac output. At the end of the 30 min basal perfusion period, hearts were challenged with a stepwise increase in preload in order to obtain maximal stroke work (volume loading). The effect of coronary perfusion on the heart's response to volume loading was highly significant: the stroke work preload relationship was significantly shifted towards higher preload values in the -CF group. Also, the maximal work produced by the heart under the experimental conditions used was lower in the -CF group. Rate of oxygen consumption of the heart increased significantly with volume loading, from a basal value of approximately 20 microl O2 min(-1) g(-1) to approx. 40 microl O2 min(-1) g(-1), but was not significantly affected by the absence of coronary perfusion. Mechanical efficiency under basal conditions was approximately 17%, but was not affected by either volume loading or coronary perfusion. Taken as a whole, these data represent direct evidence of the effect of coronary perfusion on the mechanical performance of the trout heart, but also show that these effects are limited by significant self-adjustments that occur in the heart. PMID- 14555741 TI - Electrical activity of caudal neurosecretory neurons in seawater- and freshwater adapted flounder: responses to cholinergic agonists. AB - The caudal neurosecretory system (CNSS) of the euryhaline flounder is involved in osmoregulatory responses underlying adaptation to seawater and freshwater. This study compared electrophysiological activity and responses to cholinergic agonists in the neuroendocrine Dahlgren cells in an in vitro preparation taken from fully seawater- (SWA) or freshwater-adapted (FWA) fish. Resting membrane and action potential parameters showed few differences between SWA and FWA cells. The hyperpolarisation-activated sag potential and depolarising afterpotential were present under both conditions; however, amplitude of the latter was significantly greater in SWA cells. The proportions of cells within the population exhibiting different firing patterns were similar in both adaptation states. However, bursting parameters were more variable in FWA cells, suggesting that bursting activity was less robust. The muscarinic agonist, oxotremorine, was largely inhibitory in Dahlgren cells, but increased activity in a non-Dahlgren cell population, alpha neurons. Nicotine promoted bursting activity in SWA Dahlgren cells, whereas it inhibited over half of FWA cells. PMID- 14555742 TI - Slow death in the leopard frog Rana pipiens: neurotransmitters and anoxia tolerance. AB - While frogs such as Rana temporaria are known to withstand 4-5 h anoxia at room temperature, little is known about the neurological adaptations that permit this. Previous research has shown that changes in neuroactive compounds such as glutamate and dopamine in anoxia-sensitive (mammalian) brains follow a strikingly different pattern than is observed in truly anoxia-tolerant vertebrates such as the freshwater turtle. The present study measured changes in the levels of whole brain and extracellular amino acids, and extracellular dopamine, in the normoxic and 3-4 h anoxic frog Rana pipiens, in order to determine whether their neurotransmitter responses resemble the anoxia-vulnerable or anoxia-tolerant response. Increases in whole brain serine, glycine, alanine and GABA levels were similar to those seen in anoxia-tolerant species, although the levels of glutamine, taurine and glutamate did not increase as occurs in true facultative anaerobes. Extracellular levels of aspartate, taurine and GABA also increased significantly, while glutamate levels decreased. The maintenance of low extracellular glutamate was the most significant difference between the frog and the anoxic/ischemic mammalian brain, although aspartate did increase 215% over a 4 h period of anoxia. A 12-fold increase in extracellular dopamine levels during anoxia was the biggest contrast between anoxia-tolerant vertebrates and R. pipiens. The frog could thus be an interesting model in which to examine the mechanisms of dopamine failure in early anoxia, which occurs rapidly in the mammal but over a period of hours in the 'slow death' of the anoxic frog brain. PMID- 14555744 TI - Photobehavior of stony corals: responses to light spectra and intensity. AB - Tentacle expansion and contraction were investigated in four zooxanthellate coral species and one azooxanthellate coral (Cladopsammia gracilis). Favia favus, Plerogyra sinuosa and Cladopsammia gracilis expand their tentacles at night, while tentacles in Goniopora lobata and Stylophora pistillata are expanded continuously. Light at wavelengths in the range 400-520 nm was most effective in eliciting full tentacle contraction in F. favus and in P. sinuosa. Higher light intensities in the range 660-700 nm also caused tentacle contractions in F. favus. Tentacles in C. gracilis did not respond to light. Zooxanthellar densities in tentacles were significantly higher in G. lobata, which has continuously expanded tentacles, than in F. favus and P. sinousa, where tentacles are expanded at night. Photosynthetic efficiency in F. favus and P. sinuosa was lower in specimens with contracted tentacles. However, in the dark, no differences were found in the maximum quantum yield of photochemistry in PSII (Fv/Fm) of the expanded versus the contracted tentacles of any of the four species. This work suggests that species whose tentacles remain continuously expanded have either dense algal populations in their tentacles, as in G. lobata, or minute tentacles, like S. pistillata. Dense algal populations in tentacles allow harvesting of light while small tentacles do not scatter light or shade zooxanthellae in the underlying body of the polyp. PMID- 14555743 TI - Seismic signals in a courting male jumping spider (Araneae: Salticidae). AB - Visual displays in jumping spiders have long been known to be among the most elaborate animal communication behaviours. We now show that one species, Habronattus dossenus, also exhibits an unprecedented complexity of signalling behaviour in the vibratory (seismic) modality. We videotaped courtship behaviour and used laser vibrometry to record seismic signals and observed that each prominent visual signal is accompanied by a subsequent seismic component. Three broad categories of seismic signals were observed ('thumps', 'scrapes' and 'buzzes'). To further characterize these signals we used synchronous high-speed video and laser vibrometry and observed that only one seismic signal component was produced concurrently with visual signals. We examined the mechanisms by which seismic signals are produced through a series of signal ablation experiments. Preventing abdominal movements effectively 'silenced' seismic signals but did not affect any visual component of courtship behaviour. Preventing direct abdominal contact with the cephalothorax, while still allowing abdominal movement, only silenced thump and scrape signals but not buzz signals. Therefore, although there is a precise temporal coordination of visual and seismic signals, this is not due to a common production mechanism. Seismic signals are produced independently of visual signals, and at least three independent mechanisms are used to produce individual seismic signal components. PMID- 14555745 TI - The aerodynamics of avian take-off from direct pressure measurements in Canada geese (Branta canadensis). AB - Direct pressure measurements using electronic differential pressure transducers along bird wings provide insight into the aerodynamics of these dynamically varying aerofoils. Acceleration-compensated pressures were measured at five sites distributed proximally to distally from the tertials to the primaries along the wings of Canada geese. During take-off flight, ventral-to-dorsal pressure is maintained at the proximal wing section throughout the wingstroke cycle, whereas pressure sense is reversed at the primaries during upstroke. The distal sites experience double pressure peaks during the downstroke. These observations suggest that tertials provide weight-support throughout the wingbeat, that the wingtip provides thrust during upstroke and that the kinetic energy of the rapidly flapping wings may be dissipated via retarding aerodynamic forces (resulting in aerodynamic work) at the end of downstroke. PMID- 14555746 TI - A role for nitric oxide in hypoxia-induced activation of cardiac KATP channels in goldfish (Carassius auratus). AB - Hypoxia-induced shortening of cardiac action potential duration (APD) has been attributed in mammalian hearts to the activation of ATP-sensitive potassium (KATP) channels. Since KATP channels are also present at high densities in the hearts of vertebrate ectotherms, speculation arises as to their function during periods of reduced environmental oxygen. The purpose of the present study was to determine whether nitric oxide (NO) plays a role in cardiac sarcolemmal KATP channel activation during hypoxia in a species with a high degree of tolerance to low oxygen environments: the goldfish (Carassius auratus). Conventional intracellular and patch-clamp recording techniques were used to record responses from excised ventricles or isolated ventricular myocytes and inside-out patches, respectively, from fish acclimated at 21 degrees C. During moderate, substrate free hypoxia (6.1 +/- 0.2 kPa), ventricular APD was significantly shortened at 50% and 90% of full repolarization, a response that was reversible upon reoxygenation and blocked by the KATP channel antagonist BDM. Under normoxic conditions, APD was also reduced in the presence of the NO-donor SNAP (100 micromol l(-1)). In cell-attached membrane patches, sarcolemmal KATP channel activity was enhanced after 10 min hypoxia, an effect that was reduced or eliminated by simultaneous exposure to BDM, to the guanylate cyclase inhibitor ODQ or to the NO synthase inhibitor L-NAME. In cell-free patches, KATP channel activity was abolished by 2 mmol l(-1) ATP but increased by SNAP; the cGMP analog 8-Br-cGMP (200 micromol l(-1)) also enhanced activity, an effect that was eliminated by BDM. Our data indicate that NO synthesized in cardiac myocytes could enhance sarcolemmal KATP channel activation during moderate hypoxia in goldfish. This response may serve a cardioprotective role by helping to conserve ATP or by reducing intracellular Ca2+ accumulation. PMID- 14555747 TI - Analysis of myostatin gene structure, expression and function in zebrafish. AB - Myostatin is a member of the TGF-beta family that functions as a negative regulator of skeletal muscle development and growth in mammals. Recently, Myostatin has also been identified in fish; however, its role in fish muscle development and growth remains unknown. We have reported here the isolation and characterization of myostatin genomic gene from zebrafish and analysis of its expression in zebrafish embryos, larvae and adult skeletal muscles. Our data showed that myostatin was weakly expressed in early stage zebrafish embryos, and strongly expressed in swimming larvae, juvenile and skeletal muscles of adult zebrafish. Transient expression analysis revealed that the 1.2 kb zebrafish myostatin 5' flanking sequence could direct green fluorescent protein (GFP) expression predominantly in muscle cells, suggesting that the myostatin 5' flanking sequence contained regulatory elements required for muscle expression. To determine the biological function of Myostatin in fish, we generated a transgenic line that overexpresses the Myostatin prodomain in zebrafish skeletal muscles using a muscle-specific promoter. The Myostatin prodomain could act as a dominant negative and inhibit Myostatin function in skeletal muscles. Transgenic zebrafish expressing the Myostatin prodomain exhibited no significant change in myogenic gene expression and differentiation of slow and fast muscle cells at their embryonic stage. The transgenic fish, however, exhibited an increased number of myofibers in skeletal muscles, but no significant difference in fiber size. Together, these data demonstrate that Myostatin plays an inhibitory role in hyperplastic muscle growth in zebrafish. PMID- 14555748 TI - Gait parameters in vertical climbing of captive, rehabilitant and wild Sumatran orang-utans (Pongo pygmaeus abelii). AB - Vertical climbing is central to the locomotor and foraging strategies of the great apes and, indeed, to theories about the evolution of locomotor specialisations of hominoid primates. Nevertheless, its kinematics have yet to be fully evaluated. Here, we present spatio-temporal parameters of 80 climbing sequences containing 560 limb cycles obtained from video recordings of captive, rehabilitant and wild Sumatran orang-utans (Pongo pygmaeus abelii). Gait parameters such as cycle duration, duty factor, laterality of footfalls, relative stride length and normalised speed were analysed to identify the influence of body size (through an intraspecific comparison of age/sex classes), the influence of the environment (through an intraspecific comparison of animals living in different habitats) and the influence of morphology (through an interspecific comparison with the climbing characteristics of other primates) on climbing ability in orang-utans. Results show that there are only few differences between the climbing of wild, rehabilitant and captive individuals. Cycle duration is longer and speed is lower for the wild individuals than for other groups due to the complexity of their environment and lack of familiarity with each substrate, which results in a more cautious gait. Sex/age-related differences in climbing were found to be small, although juveniles generally exhibited a shorter cycle duration and lower duty factor than other groups. The spatio-temporal gait parameters of adult females carrying infants were not found to differ significantly from those for females without an infant, which suggests that climbing kinematics are not affected by the presence of a clinging infant. Extended-elbow vertical climbing is primarily characterised by a higher duty factor than flexed-elbow climbing, indicating that the former is an energetically more demanding form of locomotion. In comparison with other primates, orang-utans exhibit a longer cycle duration, longer strides but lower climbing speed, reflecting a compromise between the demands of a large body mass and extreme joint mobility. PMID- 14555749 TI - Exposure to ultraviolet radiation causes apoptosis in developing sea urchin embryos. AB - Laboratory exposures of embryos from the sea urchin Strongylocentrotus droebachiensis to ultraviolet B radiation (UV-B, 290-320 nm), equivalent to a depth of 1-3 m in the Gulf of Maine, resulted in significant damage to DNA measured as cyclobutane pyrimidine dimer formation. Cells with DNA damage caused by ultraviolet radiation (UVR, 290-400 nm) and oxidative stress can survive, but are often retained in the G1/S phase of the cell cycle to repair DNA as a result of the expression of cell cycle genes such as p53 and p21, and the subsequent inhibition of the activity of cyclin-dependent kinases such as cdc2; if DNA cannot be repaired it can lead to programmed cell death or apoptosis. Sea urchin embryos exposed to UV-B radiation exhibit significantly higher protein concentrations of the antioxidant enzyme superoxide dismutase, and the transcriptional activators p53 and p21. The downstream activator of the cell cycle, cdc2, showed significantly lower protein concentrations with exposure to increasingly shorter wavelengths of UVR. Decreases in cdc2 could have been caused directly by exposure to UV-B or as a result of downregulation via the p53, p21 cascade, or both. These cellular events lead to apoptosis, as shown by the significant increase in DNA strand breaks observed in the nuclei of developing embryos exposed to UVR using the TUNEL assay. Cellular death, and a decrease in sea urchin embryo survivorship, are caused by the indirect and direct effects of exposure to UVR that leads to apoptosis in these laboratory experiments. PMID- 14555750 TI - Immunohistochemical fiber typing of harbor seal skeletal muscle. AB - There is strong evidence that pinnipeds maintain a lipid-based, aerobic metabolism during diving. However, the few fiber-typing studies performed on pinniped skeletal muscles are not consistent with an aerobic physiological profile. The objective of this study was to reexamine the fiber type distribution throughout the primary locomotory muscles of the harbor seal Phoca vitulina. Results from immunohistochemical (IHC) fiber typing indicated that harbor seal swimming muscles (the epaxial muscles) are composed of 47.4% type I (slow twitch, oxidative) fibers and 52.8% IIa (fast twitch, oxidative) fibers, which are homogeneously distributed throughout the muscle. Harbor seal pectoralis, a secondary swimming muscle, was composed of 16.2% type I and 84.3% type IIa fibers. No fast twitch, glycolytic (type IIb) fibers were detected in either muscle, in contrast to published data on fiber typing of harbor seal epaxial muscles using traditional histochemical techniques. The extreme specificity inherent in the IHC fiber typing procedure leads us to conclude that harbor seal swimming muscle is entirely composed of oxidative fibers. Our results are consistent with the enzymatic analyses of pinniped skeletal muscle that support the use of lipid-derived aerobic catabolism to fuel working muscle during diving in these marine mammals. PMID- 14555751 TI - Short-term transformation and long-term replacement of branchial chloride cells in killifish transferred from seawater to freshwater, revealed by morphofunctional observations and a newly established 'time-differential double fluorescent staining' technique. AB - Short- and long-term responses to direct transfer from seawater to freshwater were examined in gill chloride cells of killifish, which developed distinct freshwater- and seawater-type chloride cells in the respective environments. In a short-term response within 24 h after transfer, seawater-type chloride cells forming a pit structure on the apical surface were transformed into freshwater type cells equipped with developed microvilli on the flat or projecting apical membrane, via the intermediate type. The transformation process was accompanied by the disappearance of apically located Cl- channel (cystic fibrosis transmembrane conductance regulator) and neighboring accessory cells. Chloride cell replacement was also examined as a long-term adaptation to freshwater transfer, using a newly established 'time-differential double fluorescent staining (TDS)' technique. In the TDS technique, in vivo labeling of chloride cells was performed on two separate days, using two distinguishable mitochondria specific fluorescent probes. For 3 days after freshwater transfer, 14.7% of seawater-type cells were replaced with newly differentiated freshwater-type cells, whereas these ratios of chloride cell replacement were much lower (1.2% and 1.8%) in seawater- and freshwater-maintained groups, respectively. In consequence, following direct transfer of killifish from seawater to freshwater, seawater-type chloride cells were transformed morphologically and functionally into freshwater-type cells as a short-term response, followed by the promotion of chloride cell replacement as a long-term response. PMID- 14555752 TI - The ontogenetic scaling of hydrodynamics and swimming performance in jellyfish (Aurelia aurita). AB - It is not well understood how ontogenetic changes in the motion and morphology of aquatic animals influence the performance of swimming. The goals of the present study were to understand how changes in size, shape and behavior affect the hydrodynamics of jet propulsion in the jellyfish Aurelia aurita and to explore how such changes affect the ontogenetic scaling of swimming speed and cost of transport. We measured the kinematics of jellyfish swimming from video recordings and simulated the hydrodynamics of swimming with two computational models that calculated thrust generation by paddle and jet mechanisms. Our results suggest that thrust is generated primarily by jetting and that there is negligible thrust generation by paddling. We examined how fluid forces scaled with body mass using the jet model. Despite an ontogenetic increase in the range of motion by the bell diameter and a decrease in the height-to-diameter ratio, we found that thrust and acceleration reaction scaled with body mass as predicted by kinematic similarity. However, jellyfish decreased their pulse frequency with growth, and speed consequently scaled at a lower exponential rate than predicted by kinematic similarity. Model simulations suggest that the allometric growth in Aurelia results in swimming that is slower, but more energetically economical, than isometric growth with a prolate bell shape. The decrease in pulse frequency over ontogeny allows large Aurelia medusae to avoid a high cost of transport but generates slower swimming than if they maintained a high pulse frequency. Our findings suggest that ontogenetic change in the height-to-diameter ratio and pulse frequency of Aurelia results in swimming that is relatively moderate in speed but is energetically economical. PMID- 14555753 TI - Adaptations to diving hypoxia in the heart, kidneys and splanchnic organs of harbor seals (Phoca vitulina). AB - Pinnipeds (seals and sea lions) have an elevated mitochondrial volume density [VV(mt)] and elevated citrate synthase (CS) and beta-hydroxyacyl-CoA dehydrogenase (HOAD) activities in their swimming muscles to maintain an aerobic, fat-based metabolism during diving. The goal of this study was to determine whether the heart, kidneys and splanchnic organs have an elevated VV(mt) and CS and HOAD activities as parallel adaptations for sustaining aerobic metabolism and normal function during hypoxia in harbor seals (Phoca vitulina). Samples of heart, liver, kidney, stomach and small intestine were taken from 10 freshly killed harbor seals and fixed in glutaraldehyde for transmission electron microscopy or frozen in liquid nitrogen for enzymatic analysis. Samples from dogs and rats were used for comparison. Within the harbor seal, the liver and stomach had the highest VV(mt). The liver also had the highest CS activity. The kidneys and heart had the highest HOAD activities, and the liver and heart had the highest lactate dehydrogenase (LDH) activities. Mitochondrial volume densities scaled to tissue-specific resting metabolic rate [VV(mt)/RMR] in the heart, liver, kidneys, stomach and small intestine of harbor seals were elevated (range 1.2-6.6x) when compared with those in the dog and/or rat. In addition, HOAD activity scaled to tissue-specific RMR in the heart and liver of harbor seals was elevated compared with that in the dog and rat (3.2x and 6.2x in the heart and 8.5x and 5.5x in the liver, respectively). These data suggest that organs such as the liver, kidneys and stomach possess a heightened ability for aerobic, fat based metabolism during hypoxia associated with routine diving. However, a heightened LDH activity in the heart and liver indicates an adaptation for the anaerobic production of ATP on dives that exceed the animal's aerobic dive limit. Hence, the heart, liver, kidneys and gastrointestinal organs of harbor seals exhibit adaptations that promote an aerobic, fat-based metabolism under hypoxic conditions but can provide ATP anaerobically if required. PMID- 14555754 TI - Waved albatrosses can navigate with strong magnets attached to their head. AB - The foraging excursions of waved albatrosses Phoebastria irrorata during incubation are ideally suited for navigational studies because they navigate between their Galapagos breeding site and one specific foraging site in the upwelling zone of Peru along highly predictable, straight-line routes. We used satellite telemetry to follow free-flying albatrosses after manipulating magnetic orientation cues by attaching magnets to strategic places on the birds' heads. All experimental, sham-manipulated and control birds, were able to navigate back and forth from Galapagos to their normal foraging sites at the Peruvian coast over 1000 km away. Birds subjected to the three treatments did not differ in the routes flown or in the duration and speed of the trips. The interpretations and implications of this result depend on which of the current suggested magnetic sensory mechanisms is actually being used by the birds. PMID- 14555755 TI - Energy metabolism and valve closure behaviour in the Asian clam Corbicula fluminea. AB - Since its invasion of Europe in the early 1980s, the Asian clam Corbicula fluminea has become very abundant in nearly all western river systems. Today this species is one of the most important biomass producers in the River Rhine. Monitoring the valve movements of C. fluminea over a period of 2 years revealed a circadian rhythm in summer, with extended periods (10-12 h) of valve closure, predominantly in the morning hours. Altogether valve movements were very scarce, frequently fewer than four movements per individual per day. Simultaneous measurements of heat dissipation and oxygen consumption (calorespirometry) revealed an intermittent metabolism in the clam. With the onset of valve closure, C. fluminea reduced its metabolic rate to 10% of the standard metabolic rate (SMR) measured when the valves were open. Nevertheless, this depressed metabolism remained aerobic for several hours, enabling the clam to save energy and substrates compared to the requirements of the tenfold higher SMR. Only during long-lasting periods of valve closure (more than 5-10 h) did the clams become anaerobic and accumulate succinate within their tissues (2 micromol g(-1) fresh mass). Succinate is transported into the mantle cavity fluid, where it reaches concentrations of 4-6 mmol l(-1). Because this succinate-enriched fluid must pass the gills when the valves open again, we suggest that this anaerobic end product is at least partly reabsorbed, thus reducing the loss of valuable substrates during anaerobiosis. Propionate was also produced, but only during experimental N2-incubation, under near-anoxic conditions. The intermittent metabolism of C. fluminea is discussed as an adaption to efficiently exploit the rare food supply, saving substrates by the pronounced metabolic depression during valve closure. PMID- 14555756 TI - Developmental amnesia and its relationship to degree of hippocampal atrophy. AB - Two groups of adolescents, one born preterm and one with a diagnosis of developmental amnesia, were compared with age-matched normal controls on measures of hippocampal volume and memory function. Relative to control values, the preterm group values showed a mean bilateral reduction in hippocampal volume of 8 9% (ranging to 23%), whereas the developmental amnesic group values showed a reduction of 40% (ranging from 27% to 56%). Despite equivalent IQ and immediate memory scores in the two study groups, there were marked differences between them on a wide variety of verbal and visual delayed memory tasks. Consistent with their diagnosis, the developmental amnesic group was impaired relative to both other groups on nearly all delayed memory measures. The preterm group, by contrast, was significantly impaired relative to the controls on only a few memory measures, i.e., route following and prospective memory. We suggest that early hippocampal pathology leads to the disabling memory impairments associated with developmental amnesia when the volume of this structure is reduced below normal by approximately 20-30% on each side. Whether this is a sufficient condition for the disorder or whether abnormality in other brain regions is also necessary remains to be determined. PMID- 14555758 TI - Differential regulation of symmetry genes and the evolution of floral morphologies. AB - Shifts in flower symmetry have occurred frequently during the diversification of angiosperms, and it is thought that such shifts play important roles in plant pollinator interactions. In the model developmental system Antirrhinum majus (snapdragon), the closely related genes CYCLOIDEA (CYC) and DICHOTOMA (DICH) are needed for the development of zygomorphic flowers and the determination of adaxial (dorsal) identity of floral organs, including adaxial stamen abortion and asymmetry of adaxial petals. However, it is not known whether these genes played a role in the divergence of species differing in flower morphology and pollination mode. We compared A. majus with a close relative, Mohavea confertiflora (desert ghost flower), which differs from Antirrhinum in corolla (petal) symmetry and pollination mode. In addition, Mohavea has undergone a homeotic-like transformation in stamen number relative to Antirrhinum, aborting the lateral and adaxial stamens during flower development. Here we show that the patterns of expression of CYC and DICH orthologs have shifted in concert with changes in floral morphology. Specifically, lateral stamen abortion in Mohavea is correlated with an expansion of CYC and DICH expression, and internal symmetry of Mohavea adaxial petals is correlated with a reduction in DICH expression during petal differentiation. We propose that changes in the pattern of CYC and DICH expression have contributed to the derived flower morphology of Mohavea and may reflect adaptations to a pollination strategy resulting from a mimetic relationship, linking the genetic basis for morphological evolution to the ecological context in which the morphology arose. PMID- 14555757 TI - Processive phosphorylation of alternative splicing factor/splicing factor 2. AB - SR proteins, named for their multiple arginine/serine (RS) dipeptide repeats, are critical components of the spliceosome, influencing both constitutive and alternative splicing of pre-mRNA. SR protein function is regulated through phosphorylation of their RS domains by multiple kinases, including a family of evolutionarily conserved SR protein-specific kinases (SRPKs). The SRPK family of kinases is unique in that they are capable of phosphorylating repetitive RS domains with remarkable specificity and efficiency. Here, we carried out kinetic experiments specially developed to investigate how SRPK1 phosphorylates the model human SR protein, ASF/SF2. By using the start-trap strategy, we monitored the progress curve for ASF/SF2 phosphorylation in the absence and presence of an inhibitor peptide directed at the active site of SRPK1. ASF/SF2 modification is not altered when the inhibitor peptide (trap) is added with ATP (start). However, when the trap is added first and allowed to incubate for a specific delay time, the decrease in phosphate content of the enzyme-substrate complex follows a simple exponential decline corresponding to the release rate of SRPK1. These data demonstrate that SRPK1 phosphorylates a specific region within the RS domain of ASF/SF2 by using a fully processive catalytic mechanism, in which the splicing factor remains "locked" onto SRPK1 during RS domain modification. PMID- 14555759 TI - A genetic lesion that arrests plasma cell homing to the bone marrow. AB - The coordinated regulation of chemokine responsiveness plays a critical role in the development of humoral immunity. After antigen challenge and B cell activation, the emerging plasma cells (PCs) undergo CXCL12-induced chemotaxis to the bone marrow, where they produce Ab and persist. Here we show that PCs, but not B cells or T cells from lupus-prone NZM mice, are deficient in CXCL12-induced migration. PC unresponsiveness to CXCL12 results in a marked accumulation of PCs in the spleen of mice, and a concordant decrease in bone marrow PCs. Unlike normal mice, in NZM mice, a majority of the splenic PCs are long-lived. This deficiency is a consequence of the genetic interactions of multiple systemic lupus erythematosus susceptibility loci. PMID- 14555761 TI - Trace but not delay fear conditioning requires attention and the anterior cingulate cortex. AB - Higher cognitive functions such as attention have been difficult to model in genetically tractable organisms. In humans, attention-distracting stimuli interfere with trace but not delay conditioning, two forms of associative learning. Attention has also been correlated with activation of anterior cingulate cortex (ACC), but its functional significance is unclear. Here we show that a visual distractor interferes selectively with trace but not delay auditory fear conditioning in mice. Trace conditioning is associated with increased neuronal activity in ACC, as assayed by relative levels of c-fos expression, and is selectively impaired by lesions of this structure. The effects of the ACC lesions are unlikely to be caused by indirect impairment of the hippocampus, which is required for mnemonic aspects of trace conditioning. These data suggest that trace conditioning may be useful for studying neural substrates of attention in mice, and implicate the ACC as one such substrate. PMID- 14555760 TI - Methylated DNA-binding domain 1 and methylpurine-DNA glycosylase link transcriptional repression and DNA repair in chromatin. AB - The methyl-CpG dinucleotide containing a symmetrical 5-methylcytosine (mC) is involved in gene regulation and genome stability. We report here that methylation mediated transcriptional repressor methylated DNA-binding domain 1 (MBD1) interacts with methylpurine-DNA glycosylase (MPG), which excises damaged bases from substrate DNA. MPG itself actively represses transcription and has a synergistic effect on gene silencing together with MBD1. Chromatin immunoprecipitation analysis reveals the molecular movement of MBD1 and MPG in vivo: (i) The MBD1-MPG complex normally exists on the methylated gene promoter; (ii) treatment of cells with alkylating agent methylmethanesulfonate (MMS) induces the dissociation of MBD1 from the methylated promoter, and MPG is located on both methylated and unmethylated promoters; and (iii) after completion of the repair, the MBD1-MPG complex is restored on the methylated promoter. Mobility shift and structural analyses show that the MBD of MBD1 binds a methyl-CpG pair (mCpG x mCpG) but not the methyl-CpG pair containing a single 7-methylguanine (N) (mCpG x mCpN) that is known as one of the major lesions caused by MMS. We further demonstrate that knockdown of MBD1 by specific small interfering RNAs significantly increases cell sensitivity to MMS. These data suggest that MBD1 cooperates with MPG for transcriptional repression and DNA repair. We hypothesize that MBD1 functions as a reservoir for MPG and senses the base damage in chromatin PMID- 14555762 TI - Synchronous coadaptation in an ancient case of herbivory. AB - Coevolution has long been considered a major force leading to the adaptive radiation and diversification of insects and plants. A fundamental aspect of coevolution is that adaptations and counteradaptations interlace in time. A discordant origin of traits long before or after the origin of the putative coevolutionary selective pressure must be attributed to other evolutionary processes. Despite the importance of this distinction to our understanding of coevolution, the macroevolutionary tempo of innovation in plant defenses and insect counterdefenses has not been documented. Molecular clocks for a lineage of chrysomelid beetles of the genus Blepharida and their Burseraceae hosts were independently calibrated. Results show that these plants' defenses and the insect's counterdefensive feeding traits evolved roughly in synchrony, providing macroevolutionary confirmation of synchronous plant-herbivore coadaptation. The association between these two groups of organisms was determined to be about 112 million years old, the oldest age so far for a specialized plant-herbivore association. PMID- 14555763 TI - Multiple, recurring origins of aposematism and diet specialization in poison frogs. AB - Aposematism is the association, in a prey organism, of the presence of a warning signal with unprofitability to predators. The origin of aposematism is puzzling, because of its predicted low probability of establishment in a population due to the prey's increased conspicuousness. Aposematism is a widespread trait in invertebrate taxa, but, in vertebrates, it is mostly evident in amphibians, reptiles, and fishes. Poison frogs (Dendrobatidae) are one of the most well known examples of the co-occurrence of warning coloration and toxicity. This monophyletic group of mostly diurnal leaf-litter Neotropical anurans has both toxic/colorful and palatable/cryptic species. Previous studies suggested a single origin of toxicity and warning coloration, dividing the family in two discrete groups of primitively cryptic and more derived aposematic frogs. Recent molecular phylogenetic analyses using mostly aposematic taxa supported this conclusion and proposed a single tandem origin of toxicity and conspicuous warning coloration. By using expanded taxon and character sampling, we reexamined the phylogenetic correlation between the origins of toxicity and warning coloration. At least four or five independent origins of aposematism have occurred within poison frogs; by using simulations, we rejected hypotheses of one, two, or three origins of aposematism (P < 0.002). We also found that diet specialization is linked with the evolution of aposematism. Specialization on prey, such as ants and termites, may have evolved independently at least two times. PMID- 14555764 TI - Evolution of eusociality and the soldier caste in termites: influence of intraspecific competition and accelerated inheritance. AB - We present new hypotheses and report experimental evidence for powerful selective forces impelling the evolution of both eusociality and the soldier caste in termites. Termite ancestors likely had a nesting and developmental life history similar to that of the living family Termopsidae, in which foraging does not occur outside the host wood, and nonsoldier helpers retain lifelong options for differentiation into reproductives. A local neighborhood of families that live exclusively within a limited resource results in interactions between conspecific colonies, high mortality of founding reproductives, and opportunities for accelerated inheritance of the nest and population by offspring that differentiate into nondispersing neotenic reproductives. In addition, fertile reproductive soldiers, a type of neotenic previously considered rare and docile, frequently develop in this intraspecific competitive context. They can be highly aggressive in subsequent interactions, supporting the hypothesis that intercolonial battles influenced the evolution of modern sterile termite soldier weaponry and behaviors. PMID- 14555765 TI - AIPL1, a protein implicated in Leber's congenital amaurosis, interacts with and aids in processing of farnesylated proteins. AB - The most common form of blindness at birth, Leber's congenital amaurosis (LCA), is inherited in an autosomal recessive fashion. Mutations in six different retina specific genes, including a recently discovered gene, AIPL1, have been linked to LCA in humans. To understand the molecular basis of LCA caused by aryl hydrocarbon receptor-interacting protein-like 1 (AIPL1) mutations, and to elucidate the normal function of AIPL1, we performed a yeast two-hybrid screen using AIPL1 as bait. The screen demonstrated that AIPL1 interacts specifically with farnesylated proteins. Mutations in AIPL1 linked to LCA compromise this activity. These findings suggest that the essential function of AIPL1 within photoreceptors requires interactions with farnesylated proteins. Analysis of isoprenylation in cultured human cells shows that AIPL1 enhances the processing of farnesylated proteins. Based on these findings, we propose that AIPL1 interacts with farnesylated proteins and plays an essential role in processing of farnesylated proteins in retina. PMID- 14555766 TI - Fitness consequences of a regulatory polymorphism in a seasonal environment. AB - Gene regulation is commonly assumed to have evolved in response to environmental variability. Although tightly regulated in Escherichia coli strain K12, transcriptional control of arginine biosynthesis is deregulated in strain B. Caused by a single amino acid replacement in the arginine repressor, these contrasting regulatory strategies result in a fitness tradeoff. The K12 repressor is selectively favored in the presence of arginine and disfavored in its absence. In environments that cycle between high and low arginine, short seasons favor the K12 allele, whereas long seasons favor the B allele. Unexpectedly then, deregulated expression is adaptive in some seasonal habitats. PMID- 14555767 TI - Hepatocyte growth factor/scatter factor mediates angiogenesis through positive VEGF and negative thrombospondin 1 regulation. AB - Hepatocyte growth factor/scatter factor (HGF/SF), acting through the Met receptor, plays an important role in most human solid tumors, and inappropriate expression of this ligand-receptor pair is often associated with poor prognosis. The molecular basis for the malignant potential of the HGF/SF-Met signal in cancer cells has mostly been attributed to its mitogenic and invasive properties. However, HGF/SF also induces angiogenesis, but the signaling mechanism has not been fully explained, nor has this activity been directly associated with HGF/SF Met-mediated tumorigenesis. It is known that HGF/SF induces in vitro expression of vascular endothelial growth factor (VEGF), a key agonist of tumor angiogenesis; by contrast, thrombospondin 1 (TSP-1) is a negative regulator of angiogenesis. Here, we show that, in the very same tumor cells, in addition to inducing VEGF expression, HGF/SF dramatically down-regulates TSP-1 expression. We show that TSP-1 shut-off plays an important, extrinsic role in HGF/SF-mediated tumor development, because ectopic expression of TSP-1 markedly inhibits tumor formation through the suppression of angiogenesis. Interestingly, although VEGF induced expression is sensitive to inhibitors of several pathways, including mitogen-activated protein kinase, phosphoinositide 3-kinase, and signal transducer and activator of transcription 3, TSP-1 shut-off by HGF/SF is prevented solely by inhibiting mitogen-activated protein kinase activation. These studies identify HGF/SF as a key switch for turning on angiogenesis. They suggest that TSP-1 is a useful antagonist to tumor angiogenesis and that it may have therapeutic value when used in conjunction with inhibitors of VEGF. PMID- 14555769 TI - Agricultural ethics and multifunctionality are unavoidable. PMID- 14555770 TI - Sharing the wealth. The mechanics of a data release from industry. PMID- 14555768 TI - Treatment of Helicobacter gastritis with IL-4 requires somatostatin. AB - Fifty percent of the world's population is infected with Helicobacter pylori; however, treatment has been insufficient to eradicate the organisms due to rising antibiotic resistance. Helicobacter infection is characterized by induction of a T helper 1 lymphocyte (Th1) immune response, hypergastrinemia, and suppressed tissue somatostatin (SOM) levels. However, the mechanism by which the immune response regulates acid secretion is not known. We show here that treatment with IFN-gamma, a Th1 cytokine, was sufficient to induce gastritis, increase gastrin, and decrease SOM levels within 7 days. In contrast, the T helper 2 lymphocyte cytokine IL-4 increased SOM levels and effectively suppressed gastrin expression and secretion. This result demonstrated reciprocal regulation of acid regulatory peptides by immune modulators. IL-4 pretreatment prevented gastritis in infected wild-type but not in SOM null mice. Thus, the ability of IL-4 to oppose a Th1 mediated infection required SOM. Immunofluorescence was used to document the presence of IL-4 receptors on the gastric SOM-secreting cell (D cell). Moreover, IL-4 stimulated SOM release from primary D cell cultures. Treatment of mice chronically infected with Helicobacter felis for 2 mo with the SOM analogue octreotide resolved the inflammation. Thus, a mechanism by which IL-4 resolves inflammation in the stomach is by stimulating the release of SOM from gastric D cells. PMID- 14555771 TI - Control of ascorbate synthesis by respiration and its implications for stress responses. PMID- 14555772 TI - Chemical genetic approaches to plant biology. PMID- 14555773 TI - Is ATP a signaling agent in plants? PMID- 14555774 TI - A gateway cloning vector set for high-throughput functional analysis of genes in planta. AB - The current challenge, now that two plant genomes have been sequenced, is to assign a function to the increasing number of predicted genes. In Arabidopsis, approximately 55% of genes can be assigned a putative function, however, less than 8% of these have been assigned a function by direct experimental evidence. To identify these functions, many genes will have to undergo comprehensive analyses, which will include the production of chimeric transgenes for constitutive or inducible ectopic expression, for antisense or dominant negative expression, for subcellular localization studies, for promoter analysis, and for gene complementation studies. The production of such transgenes is often hampered by laborious conventional cloning technology that relies on restriction digestion and ligation. With the aim of providing tools for high throughput gene analysis, we have produced a Gateway-compatible Agrobacterium sp. binary vector system that facilitates fast and reliable DNA cloning. This collection of vectors is freely available, for noncommercial purposes, and can be used for the ectopic expression of genes either constitutively or inducibly. The vectors can be used for the expression of protein fusions to the Aequorea victoria green fluorescent protein and to the beta-glucuronidase protein so that the subcellular localization of a protein can be identified. They can also be used to generate promoter-reporter constructs and to facilitate efficient cloning of genomic DNA fragments for complementation experiments. All vectors were derived from pCambia T-DNA cloning vectors, with the exception of a chemically inducible vector, for Agrobacterium sp.-mediated transformation of a wide range of plant species. PMID- 14555775 TI - RNA interference in the moss Physcomitrella patens. AB - The moss Physcomitrella patens performs efficient homologous recombination, which allows for the study of individual gene function by generating gene disruptions. Yet, if the gene of study is essential, gene disruptions cannot be isolated in the predominantly haploid P. patens. Additionally, disruption of a gene does not always generate observable phenotypes due to redundant functions from related genes. However, RNA interference (RNAi) can provide mutants for both of these situations. We show that RNAi disrupts gene expression in P. patens, adding a significant tool for the study of plant gene function. To assay for RNAi in moss, we constructed a line (NLS-4) expressing a nuclearly localized green fluorescent protein (GFP):beta-glucuronidase (GUS) fusion reporter protein. We targeted the reporter protein with two RNAi constructs, GUS-RNAi and GFP-RNAi, expressed transiently by particle bombardment. Transformed protonemal cells are marked by cobombardment with dsRed2, which diffuses between the nucleus and cytoplasm. Cells transformed with control constructs have nuclear/cytoplasmic red fluorescence and nuclear green fluorescence. In cells transformed with GUS-RNAi or GFP-RNAi constructs, the nuclear green fluorescence was reduced on average 9 fold as soon as 48 h after transformation. Moreover, isolated lines of NLS-4 stably transformed with GUS-RNAi construct have silenced nuclear GFP, indicating that RNAi is propagated stably. Thus, RNAi adds a powerful tool for functional analysis of plant genes in moss. PMID- 14555776 TI - DNA sequence-based "bar codes" for tracking the origins of expressed sequence tags from a maize cDNA library constructed using multiple mRNA sources. AB - To enhance gene discovery, expressed sequence tag (EST) projects often make use of cDNA libraries produced using diverse mixtures of mRNAs. As such, expression data are lost because the origins of the resulting ESTs cannot be determined. Alternatively, multiple libraries can be prepared, each from a more restricted source of mRNAs. Although this approach allows the origins of ESTs to be determined, it requires the production of multiple libraries. A hybrid approach is reported here. A cDNA library was prepared using 21 different pools of maize (Zea mays) mRNAs. DNA sequence "bar codes" were added during first-strand cDNA synthesis to uniquely identify the mRNA source pool from which individual cDNAs were derived. Using a decoding algorithm that included error correction, it was possible to identify the source mRNA pool of more than 97% of the ESTs. The frequency at which a bar code is represented in an EST contig should be proportional to the abundance of the corresponding mRNA in the source pool. Consistent with this, all ESTs derived from several genes (zein and adh1) that are known to be exclusively expressed in kernels or preferentially expressed under anaerobic conditions, respectively, were exclusively tagged with bar codes associated with mRNA pools prepared from kernel and anaerobically treated seedlings, respectively. Hence, by allowing for the retention of expression data, the bar coding of cDNA libraries can enhance the value of EST projects. PMID- 14555778 TI - Interpreting the plastid carbon, nitrogen, and energy status. A role for PII? PMID- 14555779 TI - Biochemistry and molecular biology of antioxidants in the rhizobia-legume symbiosis. PMID- 14555780 TI - wCLUTO: a Web-enabled clustering toolkit. AB - As structural and functional genomics efforts provide the biological community with ever-broadening sets of interrelated data, the need to explore such complex information for subtle relationships expands. We present wCLUTO, a Web-enabled version of the stand-alone application CLUTO, designed to apply clustering methods to genomic information. Its first application is focused on the clustering transcriptome data from microarrays. Data can be uploaded by the user into the clustering tool, a choice of several clustering methods can be made and configured, and data are presented to the user in a variety of visual formats, including a three-dimensional "mountain" view of the clusters. Parameters can be explored to rapidly examine a variety of clustering results, and the resulting clusters can be downloaded either for manipulation by other programs or to be saved in a format for publication. PMID- 14555777 TI - Cytoskeleton-plasma membrane-cell wall continuum in plants. Emerging links revisited. PMID- 14555781 TI - Isolation of a crystal matrix protein associated with calcium oxalate precipitation in vacuoles of specialized cells. AB - The formation of calcium (Ca) oxalate crystals is considered to be a high capacity mechanism for regulating Ca in many plants. Ca oxalate precipitation is not a stochastic process, suggesting the involvement of specific biochemical and cellular mechanisms. Microautoradiography of water lettuce (Pistia stratiotes) tissue exposed to 3H-glutamate showed incorporation into developing crystals, indicating potential acidic proteins associated with the crystals. Dissolution of crystals leaves behind a crystal-shaped matrix "ghost" that is capable of precipitation of Ca oxalate in the original crystal morphology. To assess whether this matrix has a protein component, purified crystals were isolated and analyzed for internal protein. Polyacrylamide gel electrophoresis revealed the presence of one major polypeptide of about 55 kD and two minor species of 60 and 63 kD. Amino acid analysis indicates the matrix protein is relatively high in acidic amino acids, a feature consistent with its solubility in formic acid but not at neutral pH. 45Ca-binding assays demonstrated the matrix protein has a strong affinity for Ca. Immunocytochemical localization using antibody raised to the isolated protein showed that the matrix protein is specific to crystal-forming cells. Within the vacuole, the surface and internal structures of two morphologically distinct Ca oxalate crystals, raphide and druse, were labeled by the antimatrix protein serum, as were the surfaces of isolated crystals. These results demonstrate that a specific Ca-binding protein exists as an integral component of Ca oxalate crystals, which holds important implications with respect to regulation of crystal formation. PMID- 14555782 TI - Cis-acting elements and DNA-binding proteins involved in CO2-responsive transcriptional activation of Cah1 encoding a periplasmic carbonic anhydrase in Chlamydomonas reinhardtii. AB - Expression of Cah1, encoding a periplasmic carbonic anhydrase in Chlamydomonas reinhardtii Dangeard, is activated when cells are exposed to low-CO2 conditions (0.04% [v/v]) in light. By using an arylsulfatase reporter gene, a regulatory region essential for the transcriptional activation of Cah1 was delimited to a 63 bp fragment between -293 and -231 relative to the transcription start site. Linker-scan analysis of the 63-bp region identified two enhancer elements, EE-1 (AGATTTTCACCGGTTGGAAGGAGGT) and EE-2 (CGACTTACGAA). Gel mobility shift assays indicated that nuclear extracts purified from cells grown under low-CO2 conditions in light contained DNA-binding proteins specifically interacting with EE-1 and EE-2. Gel mobility shift assays using mutant oligonucleotide probes revealed that the protein binding to EE-1 preferentially recognized a 9-bp sequence stretch (AGATTTTCA) of EE-1, containing a conserved sequence motif named EEC, GANTTNC, which is also present in EE-2. The EE-1- and EE-2-binding proteins interacted with the EECs contained in both of the two enhancer elements in vitro. Four EECs in the 5'-upstream region from -651 to -231 of Cah1 played a central role in the transcriptional activation of Cah1 under low-CO2 conditions. These EEC-binding proteins were present even in cells grown under high-CO2 conditions (5% [v/v]) or in the dark when Cah1 is not activated. On the basis of these results, the relationship between the transcriptional regulation of Cah1 and protein-binding to the enhancer elements in the 5'-upstream region of Cah1 is discussed. PMID- 14555784 TI - Focus issue: cell death in the heart. PMID- 14555783 TI - Interaction of calmodulin, a sorting nexin and kinase-associated protein phosphatase with the Brassica oleracea S locus receptor kinase. AB - Recognition of self-pollen during the self-incompatibility response in Brassica oleracea is mediated by the binding of a secreted peptide (the S locus cysteine rich protein) to the S locus receptor kinase (SRK), a member of the plant receptor kinase (PRK) superfamily. Here, we describe the characterization of three proteins that interact with the cytosolic kinase domain of SRK. A B. oleracea homolog of Arabidopsis kinase-associated protein phosphatase was shown to interact with and dephosphorylate SRK and was itself phosphorylated by SRK. Yeast (Saccharomyces cerevisiae) two-hybrid screens identified two additional interactors, calmodulin and a sorting nexin, both of which have been implicated in receptor kinase down-regulation in animals. A calmodulin-binding site was identified in sub-domain VIa of the SRK kinase domain. The binding site is conserved and functional in several other members of the PRK family. The sorting nexin also interacted with diverse members of the PRK family, suggesting that all three of the interacting proteins described here may play a general role in signal transduction by this family of proteins. PMID- 14555785 TI - Apoptosis in heart failure and the senescent heart. AB - The progressive loss of cardiac myocytes by apoptotic cell death has been discussed as an important pathogenic component in the failing myocardium as well in the aging heart. The degree to which apoptosis contributes to myocyte loss in these conditions, however, is a controversial issue. This review focuses on the regulation of apoptosis, evidence implicating apoptosis as a mechanism for the progression and development of heart failure, the role of apoptotic death in senescent cardiac dysfunction, as well as on the problems of detection of apoptosis. PMID- 14555786 TI - The balance between pro-apoptotic and anti-apoptotic pathways in the failing myocardium. AB - The purpose of this review is to highlight those circulating molecules, membrane receptors, and signaling pathways that initiate, potentiate, or conversely, inhibit apoptosis within cardiomyocytes. This review focuses on pathways directly related to the failing heart and discusses the limitations of current methodologies for assessing cardiomyocellular apoptosis. It is important to note that the adrenergic, reactive oxygen species, and proinflammatory cytokine signaling pathways are not the only pro-apoptotic pathways active in the myocardium, nor are IL-6-related cytokine, calcineurin, and IGF-1/PI3K/Akt signaling pathways the only anti-apoptotic pathways active in the myocardium. However, they are among the best-characterized apoptosis-mediating pathways and therefore they may serve as foundation for future studies aimed at identifying novel apoptotic regulating pathways active in cardiomyocytes. Considering the short history of studying cardiomyocellular apoptosis, a tremendous body of knowledge has been collected. Understandably, much more work remains. Tomorrow's studies must (1) continue to examine the signaling pathways mediating cardiomyocellular apoptosis by focusing on the links to the ubiquitous apoptosis effectors, (2) use the expanding body of knowledge to develop more specific inhibitors of apoptosis, and then (3) confirm the causal relationship of cardiomyocellular apoptosis and cardiac dysfunction in physiologic models of cardiac challenge. PMID- 14555787 TI - Bringing cell death alive. AB - The role of apoptosis in ischemia and reperfusion of the heart has been widely debated. This controversy has been continued because of the lack of an apoptosis detection method that allowed obtaining detailed kinetic and quantitative information on apoptosis. Here we focus on recent findings that look into the detection of apoptosis following ischemia and reperfusion in the heart in animal models and in patients using Annexin-A5 based image technology. Following cardiac cell damage, one major characteristic finding is that apoptotic cells express phosphatidylserines (PS) on the outer leaflet of their cell membrane, serving as a "remove me" signal for the immune system. Annexin-A5, a native plasma protein with a high affinity for PS, can be used to measure this mode of cell death. Several Annexin-A5 based imaging systems have been developed to measure apoptosis from cell culture up to patients. In this review, implications, limitations, and clinical relevance of cell death imaging will be discussed. PMID- 14555788 TI - Cell death and diabetic cardiomyopathy. AB - Myocardial cell death is a key element in the pathogenesis and progression of various etiological cardiomyopathies such as ischemia-reperfusion, toxic exposure, and various chronic diseases including myocardial infarction, atherosclerosis, and endothelial dysfunction. Myocardial cell death is also observed in the hearts of diabetic patients and animal models; however, its importance in the development of diabetic cardiomyopathy is not completely understood. The goal of this review is to summarize our current understanding of the characteristics of diabetes-induced myocardial cell death. In the search of the mechanisms by which diabetes induces myocardial cell death, multiple cell death pathways have been proposed. Reactive oxygen and nitrogen species accumulation plays a critical role in the cell death process. Several studies have shown that suppression of myocardial cell death by antioxidants or inhibitors for apoptosis-specific signaling pathways results in a significant prevention of diabetic cardiotoxicity, suggesting that cell death in diabetic subjects plays an important role in the development of diabetic cardiomyopathy. PMID- 14555789 TI - NF-kappaB as an integrator of diverse signaling pathways: the heart of myocardial signaling? AB - NF-kappaB is a pleiotropic transcription factor implicated in the regulation of diverse biological phenomena, including apoptosis, cell survival, cell growth, cell division, innate immunity, cellular differentiation, and the cellular responses to stress, hypoxia, stretch and ischemia. In the heart, NF-kappaB has been shown to be activated in atherosclerosis, myocarditis, in association with angina, during transplant rejection, after ischemia/reperfusion, in congestive heart failure, dilated cardiomyopathy, after ischemic and pharmacological preconditioning, heat shock, burn trauma, and in hypertrophy of isolated cardiomyocytes. Regulation of NF-kappaB is complicated; in addition to being activated by canonical cytokine-mediated pathways, NF-kappaB is activated by many of the signal transduction cascades associated with the development of cardiac hypertrophy and response to oxidative stress. Many of these signaling cascades activate NF-kappaB by activating the IkappaB kinase (IKK) complex a major component of the canonical pathway. These signaling interactions occur largely via signaling crosstalk involving the mitogen-activated protein kinase/extracellular signalregulated kinase kinases (MEKKs) that are components of mitogen activated protein kinase (MAPK) signaling pathways. Additionally, there are other signaling factors that act more directly to activate NF-kappaB via IkappaB or by direct phosphorylation of NF-kappaB subunits. Finally, there are combinatorial interactions at the level of the promoter between NF-kappaB, its coactivators, and other transcription factors, several of which are activated by MAPK and cytokine signaling pathways. Thus, in addition to being a major mediator of cytokine effects in the heart, NF-kappaB is positioned as a signaling integrator. As such, NF-kappaB functions as a key regulator of cardiac gene expression programs downstream of multiple signal transduction cascades in a variety of physiological and pathophysiological states. We show that genetic blockade of NF-kappaB reduces infarct size in the murine heart after ischemia/reperfusion (I/R), implicating NF-kappaB as a major determinant of cell death after I/R. These results support the concept that NF-kappaB may be an important therapeutic target for specific cardiovascular diseases. PMID- 14555790 TI - The cardioprotective agent urocortin induces expression of CT-1. AB - The corticotrophin-releasing hormone-related factor, urocortin (Ucn) and the interleukin (IL)-6 family cytokine cardiotrophin-1 (CT-1) are both cardioprotective agents able to protect the heart from ischemic damage. In both cases the protective effect involves activation of the p42/p44 MAPK and PI-3 kinase/Akt pathways, but the protective effect of Ucn requires de novo protein synthesis whereas that of CT-1 does not. In this study, we show that Ucn induces enhanced expression of CT-1 at both the mRNA and protein levels. This effect is mediated by activation of the CT-1 gene promoter and requires the transcription factor C/EBPbeta/NF-IL6. Hence, a specific cardioprotective factor can induce enhanced expression of another cardioprotective factor belonging to an unrelated protein family. PMID- 14555792 TI - The regulation and consequences of immune-mediated cell death in atheromatous diseases. AB - Atheromatous diseases are lipid and cell-rich vascular disorders that include coronary artery disease (CAD), transplant vascular disease (TVD), and restenosis. Considering the inflammatory nature of these diseases, cytotoxic immune mechanisms such as the FasL and granzyme/perforin pathways most likely play important roles in the development and remodeling of many lesions. Furthermore, although the contributions of immune responses to each disease vary, the correspondent localization of certain mediators and effectors suggests that they may contribute to a spectrum of atheromatous diseases. In this review, the contribution of immune cell-mediated cell death in the onset and pathogenesis of CAD and TVD is examined. PMID- 14555791 TI - HSP60, Bax, and cardiac apoptosis. AB - HSP60 has long been known as an important chaperonin and as having key folding functions within the mitochondria. However, it has now become evident that significant amounts of HSP60 are found in extra-mitochondrial locations. This extra-mitochondrial HSP60 in the heart has key anti-apoptotic functions. Extra mitochondrial HSP60 complexes with both bax and bak, but not with bcl-2. Reduction in HSP60 is sufficient to precipitate apoptosis. In the setting of hypoxia and reoxygenation HSP60 decreases with reoxygenation, but the apoptotic cascade has already been triggered by end-hypoxia. Redistribution of cytosolic HSP60 to the plasma membrane during hypoxia appears to contribute to the initiation of the apoptotic cascade with hypoxia and reoxygenation. PMID- 14555793 TI - Therapeutic angiogenesis: a complex problem requiring a sophisticated approach. AB - Blood and vascular disorders underlie a plethora of pathologic conditions and are the single most frequent cause of human disease. Ischemia, involving restricted blood flow to tissues is the most common consequence of vessel dysfunction resulting in the disruption of oxygen and nutrient delivery and the accumulation of waste metabolites. Cells cannot survive extended severe ischemia but may be able to adapt to a moderate condition where diffusion to and from bordering nonischemic regions sustains vital functions. Under this condition, the secondary functions of effected cells are likely to be impaired, and a new metabolic equilibrium is established, determined by the level of cross-diffusion and degree of hypoxia. In tissues with a normally high metabolic turnover such as skeletal and cardiac muscle, even mild ischemia causes hypoxia, acidosis, and depressed function (contractility) and eventually threatens myocyte viability and organ function. Ischemic cardiac muscle is additionally vulnerable because reperfusion is essential for survival but reperfusion itself poses additional stress principally from increased production of free radicals during reoxygenation. The latter effect is called reperfusion injury and can cause as much damage as the ischemia. The treatment possibilities for ischemia-related vascular disease are limited. Lipid/cholesterol-lowering agents, diet and antiplatelet adherence (aspirin) therapy may help slow the progression of vessel disease in some instances; but surgical reconstruction may be the only option in advanced stages, and even this is not always an option. An alternative and rather obvious strategy to treat ischemia is to activate endogenous angiogenic or arteriogenic pathways to stimulate revascularization of the tissue. The feasibility of such a strategy has now been established through the results of studies over the past decade, and a new discipline called therapeutic angiogenesis has emerged. This review focuses on the application of therapeutic angiogenesis for treating ischemic muscle disease and includes a critical evaluation of the parameters and limitations of current procedures. The development of this technology has benefited from its application to both peripheral and coronary artery disease and results from both are reviewed here. PMID- 14555794 TI - Astrocyte-mediated methylmercury neurotoxicity. AB - Methylmercury (MeHg) is a potent neurotoxicant. Any source of environmental mercury represents a potential risk for human MeHg poisoning, because the methylation of inorganic mercury to MeHg in waterways results ultimately in its accumulation in the sea food chain, which represents the most prevalent source for human consumption. A small amount of MeHg accumulates in the central nervous system (CNS), particularly in astrocytes. Astrocytic swelling, excitatory amino acid (EAA) release and uptake inhibition, as well as EAA transporter expression inhibition are known sequelae of MeHg exposure. Herein, we review the effect of MeHg on additional transport systems (for cystine and cysteine) as well as arachidonic acid (AA) release and cytosolic phospholipase A2 (cPLA2) regulation and attempt to integrate the effects of MeHg in astrocytes within a mechanistic hypothesis that explains the inability of these cells to maintain control of the proper milieu of the extracellular fluid and, in turn, leads to neuronal demise. PMID- 14555795 TI - Concentrations of copper, zinc, and magnesium in sera from patients with idiopathic dilated cardiomyopathy. AB - Trace elements are known to have a key role in myocardial metabolism. The accumulation (cobalt, arsenic, copper) or deficiency (selenium, zinc) of trace elements may be responsible for idiopathic dilated cardiomyopathy. We investigated the trace element concentrations (Cu, Zn, Mg) in sera from patients with dilated cardiomyopathy by atomic absorption spectrophotometry. We observed that patients with dilated cardiomyopathies have higher copper and lower zinc concentrations in serum than healthy controls. The magnesium concentrations of patients did not differ significantly from that of control subjects. PMID- 14555796 TI - Iron concentrations in intestinal cancer tissue and in colon and rectum polyps. AB - A prospective randomized trial was used to determine iron concentrations in intestinal cancer tissue and colorectum polyps. We investigated the possible difference between the concentrations of iron, ferritin, albumin, and hemoglobin in the serum of patients with colorectal cancer and polyps. We also determined the relationship between the iron and ferritin levels in cancer tissue, the localization of neoplasms, and the stage of their development. The study comprises 67 patients with colorectum cancer and 42 patients with colon and rectum polyps. The metal was determined by using the total-reflection X-ray fluorescence (TRXRF) method. The mean concentration of iron in colorectal cancer equaled 46.1 microg/g of the tissue and was higher than in the case of polyps (43.2 microg/g). The mean serum iron level in patients with colorectal cancer was statistically lower than in the serum of patients with polyp and in the control group (54.5, 91.3, and 108.0 microg/g, respectively). The determined average concentration of ferritin in the serum of patients with colorectal cancer equaled 60.4 microg/g and was statistically lower than the level of this enzyme in the serum of patients with polyps (85.2 microg/g) and in the control group (102.0 microg/g). There was no difference between the serum albumin and hemoglobin concentrations in patients with colorectal cancer, polyps, and the control. There was no difference in the levels of iron and ferritin depending on the location of the neoplasm and the stage of its development. Also, there was no difference between the concentrations of iron in the cancer tissue of malignant and benign tumors after taking into consideration sex and age of patients. During the examination we determined significantly higher concentrations of iron in the cancer tissue and not in the polyp. The low levels of iron in the serum of patients with malignant tumor may increase colorectal cancer risk. PMID- 14555798 TI - Computer simulation for effect of Pr(III) on Ca(II) speciation in human interstitial fluid. AB - A multiphase model of metal ion speciation in human interstitial fluid was constructed and the effect of Pr(III) on Ca(II) speciation was studied. Results show that free Ca2+, [Ca(HCO3)], and [Ca(Lac)] are the main species of Ca(II). Because of the competition of Pr(III) for ligands with Ca(II), the percentages of free Ca2+, [Ca(Lac)], and [Ca(His)(Thr)H3] increase gradually and the percentages of CaHPO4(aq) and [Ca(Cit)(His)H2] decrease gradually with the increase in the total concentration of Pr(III). However, the percentages of [Ca(HCO3)] and CaCO3(aq) first increase and then begin to decrease when the total concentration of Pr(III) exceeds 6.070 x 10-4 M. PMID- 14555797 TI - Plasma malondialdehyde and serum trace element concentrations in patients with active pulmonary tuberculosis. AB - Increased amounts of reactive oxygen species (ROS) are produced as a consequence of a phagocyte respiratory burst during pulmonary inflammation. The aim of our study was to assess the concentration of malondialdehyde (MDA) and trace metals in patients with active pulmonary tuberculosis (TB). Eighty-three subjects were enrolled into the study and prospectively divided into three groups: 22 subjects with healthy controls (group I), 21 patients with inactive pulmonary TB (group II), and 40 patients with active pulmonary TB (group III). Before beginning the therapy, plasma MDA and serum concentrations of zinc (Zn), copper (Cu), albumin, and iron (Fe) were measured. The concentration of MDA and Cu in group III were higher than in the other groups (p<0.0001). The serum Zn and albumin levels were significantly lower in group III compared with healthy controls (p<0.05). There was a positive correlation between MDA and erythrocyte sedimentation rate (r = +0.647, p<0.0001; Spearman's test). Our data indicated increased circulating levels of MDA and changed serum trace metal levels in active pulmonary TB. Trace metal levels must be closely followed during the diseases process and further studies are needed to assess the role of antioxidants as adjuvant therapy in patients with active pulmonary TB. PMID- 14555799 TI - Variation in the distribution of trace elements in hepatoma. AB - There are many reports of reduction of zinc level and rise of copper level in serum of patients with liver disease. However, there are a few reports that compare the trace elements in tumor tissues and nontumor tissues of the liver with hepatoma. We studied trace element distribution in tumor tissues and nontumor tissues of liver with hepatoma and compared them with data from normal liver tissues. Zinc (Zn), copper (Cu), selenium (Se), cadmium (Cd), mercury (Hg), and iron (Fe) were chosen as the trace elements to be observed. We observed falls of Zn, Cd, and Hg levels in tumor tissues and the rise of Cu level as a result of this investigation. Zn, Cd, and Hg levels in tumor tissues were significantly lower than those in nontumor tissues and Zn, Cd, and Hg levels in nontumor tissues were significantly lower than in normal liver tissues. This tendency was clearer for Cd and Hg than for Zn. Although the distribution of Cu was not significant, a distribution contrary to that of Zn was shown. These findings indicate that the distribution of Zn, Cd, and Hg can serve as supportive evidence that could be useful as a tumor marker. Selenium showed almost the same accumulation tendency among tumor tissues, nontumor tissues, and normal livers. Although correlation was observed among most metals in the normal liver, there was almost no correlation in tumor tissues. PMID- 14555800 TI - Changes in serum selenium, copper, zinc levels and cu/zn ratio in patients with pulmonary tuberculosis during therapy. AB - The effectiveness and success of antituberculosis therapy is mainly measured by its ability to identify the organism in the sputum. In certain cases, available tuberculosis tests are not satisfactory and do not provide enough information on the effectiveness of antituberculosis therapy. Copper (Cu), zinc (Zn), and selenium (Se) are the essential elements that play a crucial role in the immune system. The serum levels of these elements vary in many diseases including tuberculosis. In this study, we investigate whether the serum levels of Cu, Zn, and Se change during antituberculosis therapy. We have included 22 pulmonary tuberculosis cases that were newly diagnosed with positive sputum and 18 healthy subjects. At the beginning and 2 mo after therapy, serum levels of Cu, Zn, and Se were measured by atomic absorption spectrometry. Despite Se and Cu levels not being affected during the treatment, we found that there was a significant increase in the levels of Zn and a decrease in the Cu/Zn ratio. Serum Zn levels and the Cu/Zn ratio could be used as a valuable laboratory tool for the clinicians to assess response to therapy or effectiveness of the ongoing antituberculosis therapy. PMID- 14555801 TI - Effects of vanadyl sulfate on kidney in experimental diabetes. AB - The aim of this work was to investigate the biochemical and histological effects of vanadyl sulfate on blood glucose, urea, and creatinine in serum and nonenzymatic glycosylation and glutathione levels in kidney tissue of normal and streptozotocin (65 mg/kg) diabetic rats. Vanadyl sulfate was administered by gavage at a dose of 100 mg/kg. After 60 d of treatment, serum urea, creatinine, and blood glucose levels significantly increased in the diabetic group but not so in the vanadyl sulfate, which showed significantly reduced serum urea and blood glucose levels and a nonsignificant reduction of serum creatinine levels. Nonenzymatic glycosylation was increased and the glutathione level was decreased in the kidney tissue of diabetic rats. Treatment with vanadyl sulfate reversed these effects. Degenerative changes were detected in diabetic animals by electron and light microscopy. Although there are individual differences in diabetic animals given vanadium, some reduction of degenerative changes were observed. PMID- 14555802 TI - Transmission electron microscopy study of the effects of zinc overdose on mouse liver tissue. AB - Thirty adult male mice were divided into three groups. The animals in group I were used as controls and drank only water during the entire period of experimentation. Group II animals drank water containing 1.5 g/100 mL zinc as ZnSO4, and group III animals received 2.5 g/100 mL zinc. After 3 wk supplementation with high doses of zinc, the animals were killed and the livers were removed and examined by electron microscopic techniques. After the supplementation period, the animals in groups II and II showed various degrees of degenerative changes in the hepatocytes, such as increased size and the presence of spaces and an abundance of lipid globules in the cytoplasm. The mitochondria showed a crystalline appearance, a diluted matrix, and dense aggregations. Some smooth endoplasmic reticulum tubules showed dilation and were filled with a dense substance. None of these changes were present in the group I control animals. PMID- 14555803 TI - Approaches to immune reconstitution in HIV infection. PMID- 14555804 TI - Antiretroviral therapy: select lessons from recent studies. PMID- 14555805 TI - The increasing importance of sexually transmitted diseases in HIV-infected persons. PMID- 14555806 TI - [Control of morphogenesis of woody plant by gravity on earth]. AB - Using the weeping branch of Japanese flowering cherry tree and its woody stem of the seedling grown under simulated microgravity condition by three dimensional clinostat, it was elucidated that the morphogenesis of its secondary xylem supporting the plant itself to grow upward is seriously controlled by gravity on earth with a sedimentable amyloplast as its sensor. Space experiment of woody plant is expected to elucidate such problem. PMID- 14555807 TI - [Preface to special issue: "Molecular mechanism of the adaptation of terrestrial plants to gravity environment on Earth"]. AB - Organisms borne in the primitive sea about 30 million years ago had evolved in water without a large influence of gravity on earth. About 4 million years ago, the first terrestrial organisms, plants appeared on the land from the sea. The terrestrial plants have adapted to and evolved on the land environment so that they can extend their roots downward in soil and their shoots upward against 1 g gravity. At least two functions that were acquired during the process of evolution helped the terrestrial plants to adapt to gravity environment on earth. One is gravitropism. The other is the reinforcement of the cell wall, particularly the secondary cell wall. In the present feature articles, the molecular mechanism of the adaptation of terrestrial plants to gravity environment on earth will be reviewed, paying special attention to the mechanism of the genetic control of the signaling of gravity stimulus in gravitropism, automorphogenesis, genes involved in auxin transport, gravity effect on cell wall properties and gravimorphogenesis in terrestrial plants. PMID- 14555808 TI - [Mechanism of gravi-sensing and -transduction in gravitropism of higher plants]. AB - In higher plants, some organs such as roots, hypocotyls, and stems, can sense the direction of gravity to regulate their orientation. Gravitropic response is composed of four steps; 1. gravity sensing and conversion of physical stimuli to biochemical signals, 2. intracellular signal transduction in gravity sensing cells, 3. signal transmitting to responding tissues, 4. differential growth of organs. Here we focus on the former two steps. Recent studies using modern technique have gradually unveiled early events and mechanism of gravitropic response. Genetic approach provided evidences that strongly support the classical theory for gravity sensing (step 1). Computational analysis suggested the existence of another gravity sensing mechanism in roots. Spatial and temporal ion imaging in living organs in real time provided information on step 2. In addition, reverse genetic approach suggested asymmetrical intracellular distribution of auxin transporter [correction of transpoter] is a possible link between step 2 and 3. However, molecular basis of the signaling mechanism remains unknown. We believe extensive molecular genetic approach combined with recent techniques cited here shed the light to this ambiguous area of research. PMID- 14555809 TI - [Graviresponse in higher plants and its regulation in molecular bases: relevance to growth and development, and auxin polar transport in etiolated pea seedlings]. AB - We review the graviresponse under true and simulated microgravity conditions on a clinostat in higher plants, and its regulation in molecular bases, especially on the aspect of auxin polar transport in etiolated pea (Pisum sativum L. cv. Alaska) seedlings which were the plant materials subjected to STS-95 space experiments. True and simulated microgravity conditions substantially affected growth and development in etiolated pea seedlings, especially the direction of growth of stems and roots, resulting in automorphosis. In etiolated pea seedlings grown in space, epicotyls were the most oriented toward the direction far from the cotyledons, and roots grew toward the aerial space of Plant Growth Chamber. Automorphosis observed in space were well simulated by a clinorotation on a 3 dimensional clinostat and also phenocopied by the application of auxin polar transport inhibitors of 2,3,5-triiodobenzoic acid, N-(1-naphtyl)phthalamic acid and 9-hydroxyfluorene-9-carboxylic acid. Judging from the results described above together with the fact that activities of auxin polar transport in epicotyls of etiolated pea seedlings grown in space substantially were reduced, auxin polar transport seems to be closely related to automorphosis. Strenuous efforts to learn in molecular levels how gravity contributes to the auxin polar transport in etiolated pea epicotyls resulted in successful identification of PsPIN2 and PsAUX1 genes located in plasma membrane which products are considered to be putative efflux and influx carriers of auxin, respectively. Based on the results of expression of PsPIN2 and PsAUX1 genes under various gravistimulations, a possible role of PsPIN2 and PsAUX1 genes for auxin polar transport in etiolated pea seedlings will be discussed. PMID- 14555810 TI - [Auxin-mediated response of cucumber seedlings to gravity]. AB - Gravity regulates peg formation because cucumber seedlings grown in a horizontal position develop a peg on the lower side of the transition zone (TR zone) but not on the upper side. Studies on peg formation have suggested the regulation of peg formation by gravity as follows. Cucumber seedlings potentially develop a peg on both the lower and upper sides of the TR zone. The development of the peg on upper side of the TR zone is suppressed in response to gravity. A phytohormone, auxin, induces peg formation. Upon gravistimulation the auxin concentration on the upper side of the TR zone is reduced to a level below the threshold value necessary for peg formation. The unequally distributed auxin across TR zone is caused by a change in accumulation of auxin influx carrier (CsAUX1) protein and auxin efflux carrier (CsPIN1) protein in response to gravity. In addition, TR zone before peg initiation expresses both CsARF2 (putative activator of auxin response factor) and CsIAA1 (putative repressor of auxin-inducible gene expression), by which TR zone could respond the auxin gradient regulated by gravity. PMID- 14555811 TI - [Gravity resistance, another graviresponse in plants--function of anti gravitational polysaccharides]. AB - The involvement of anti-gravitational polysaccharides in gravity resistance, one of two major gravity responses in plants, was discussed. In dicotyledons, xyloglucans are the only cell wall polysaccharides, whose level, molecular size, and metabolic turnover were modified under both hypergravity and microgravity conditions, suggesting that xyloglucans act as anti-gravitational polysaccharides. In monocotyledonous Poaceae, (1-->3),(1-->4)-beta glucans, instead of xyloglucans, were shown to play a role as anti-gravitational polysaccharides. These polysaccharides are also involved in plant responses to other environmental factors, such as light and temperature, and to some phytohormones, such as auxin and ethylene. Thus, the type of anti-gravitational polysaccharides is different between dicotyledons and Poaceae, but such polysaccharides are universally involved in plant responses to environmental and hormonal signals. In gravity resistance, the gravity signal may be received by the plasma membrane mechanoreceptors, transformed and transduced within each cell, and then may modify the processes of synthesis and secretion of the anti gravitational polysaccharides and the cell wall enzymes responsible for their degradation, as well as the apoplastic pH, leading to the cell wall reinforcement. A series of events inducing gravity resistance are quite independent of those leading to gravitropism. PMID- 14555812 TI - Multiplex PCR for the detection of genes encoding aminoglycoside modifying enzymes and methicillin resistance among Staphylococcus species. AB - We developed multiplex polymerase chain reaction (PCR) to detect aac(6 ')/aph(2 "), aph(3 ')-IIIa, and ant(4 ')-Ia, the genes encoding the most clinically relevant amino-glycoside modifying enzymes (AME), and simultaneously, the methicillin resistant gene, mecA, in Staphylococcus species. Clinical isolates of 45 S. aureus and 47 coagulase negative staphylococci (CNS) from tertiary university hospitals were tested by conventional susceptibility testing, using the agar dilution method and by multiplex PCR. Of a total of 92 isolates, 61 isolates were found to be methicillin-resistant. Of these, 54 isolates (89%) were found to be harboring mecA. Seventy-five percent of the 92 isolates demonstrated resistance to at least one of the aminoglycosides tested. Moreover, resistance to aminoglycosides was closely associated with methicillin-resistance (p<0.05). The most prevalent AME gene was aac(6 ')/aph(2 ") which was found in 65% of the isolates, and ant(4 ')-Ia and aph(3 ')-IIIa were present in 41% and 9% of the isolates, respectively. The concordance between methicillin-resistance and the presence of mecA gene was 98% in S. aureus and 81% in CNS. The concordance between gentamicin resistance and the presence of aac(6 ')/aph(2 ") gene was 100% in S. aureus and 85% in CNS. The multiplex PCR method that we developed appears to be both a more rapid and reliable than conventional method. PMID- 14555813 TI - Detrimental effects of N(omega) nitro-L-arginine methyl ester (L-NAME)in experimental Escherichia coli sepsis in the newborn piglet. AB - The role of nitric oxide during neonatal sepsis is complex. We tested the hypothesis that nonselective inhibition of nitric oxide synthase with N(omega) nitro-L-arginine methyl ester (L-NAME) is detrimental during the early phase of experimental sepsis in the newborn piglet. Newborn piglets were divided into four groups: 6 in the control group, 6 in the L-NAME control group, 12 in the sepsis group (SG), and 11 in the sepsis with L-NAME group (NS). Sepsis was induced by intravenous injection of 10(8) colony forming units of Escherichia coli. L-NAME 10 mg/kg was given intravenously 60 min before the induction of sepsis. The survival rate of piglets after 4 hr was 27% in NS, while it was 100% in other groups. Systemic hypotension, observed in both SG and NS, were more profound in NS. Leukopenia was observed in both SG and NS. Thrombocytopenia, prolongation of prothrombin time and activated partial thromboplastin time, and increase in thrombin-antithrombin complexes were observed only in NS. Decreased PaO2 /FiO2 ratio, arterial pH and base excess, and increased blood lactate levels observed in both SG and NS, but were more profound in NS. These findings suggest that nonselective inhibition of nitric oxide synthase with L-NAME is detrimental during the early phase of experimental neonatal sepsis. PMID- 14555814 TI - Estrogen and enalapril attenuate the development of right ventricular hypertrophy induced by monocrotaline in ovariectomized rats. AB - The present study evaluated the importance of ovarian functions and the renin angiotensin system in the progression of the right ventricular (RV) hypertrophy. Female Sprague-Dawley rats were bilaterally ovariectomized (Ovx) and injected with monocrotaline (MCT, 60 mg/kg, sc). Four weeks after MCT-treatment, only the male and Ovx female rats showed marked RV hypertrophy. The hypertrophied RV of the male-MCT and Ovx-MCT rats exhibited remarkably elevated renin mRNA levels. Gene expression levels of angiotensinogen, TGF-beta1, and endothelin-1 in the hypertrophied RV also increased, but to the less degree than did the renin mRNA. To investigate beneficial effects of estrogen or enalapril on progression of the pulmonary hypertension and RV hypertrophy, histological changes of the lung and heart were examined. Sham-MCT female rats showed histological changes indicating pulmonary hypertension without RV hypertrophy. In contrast, Ovx-MCT rats showed marked RV hypertrophy with pathological changes, denoting severe pulmonary and myocardial injuries. Estrogen-or enalapril-treated Ovx-MCT rats did not show RV hypertrophy, and showed remarkably ameliorated ultrastructural changes in the lung and RV. These results from this rat model suggest that both estrogen and inhibition of the renin-angiotensin system have protective functions against the development of the pulmonary hypertension and cardiac remodeling. PMID- 14555815 TI - Effects of N-acetyl-L-cysteine and glutathione on antioxidant status of human serum and 3T3 fibroblasts. AB - The effectiveness of several sulfhydryl compounds in the treatment of paraquat intoxication has been previously tested based on their antioxidant ability. However, practical guidelines for their clinical use remain to be determined. As a preliminary pharmacokinetic study on sulfhydryl compounds, we attempted to establish the optimal concentration of N-acetyl-L-cysteine, glutathione, superoxide dismutase, and catalase. We measured the antioxidant effect of these antioxidants in normal pooled plasma and on intracellular reactive oxygen species (ROS) induced by paraquat. N-acetyl-L-cysteine begins to suppress the production of ROS in plasma at concentrations as low as 5 mM, with the suppression being maximal at 40 mM. In the same way, glutathione increased the total antioxidant status in plasma at concentrations of 5-40 mM in a dose-dependent manner. Complete suppression of ROS in plasma induced by exposure to 500 micro M paraquat for 40 min was observed when using 40 mM N-acetyl-L-cysteine and 5 mM glutathione. These concentrations are comparable with 50 units of catalase, which reduced ROS at concentrations of 5-100 units. Further pharmacokinetic study into the systemic administration of these antioxidants is necessary, using effective concentrations of 5-40 mM for both N-acetyl-L-cysteine and glutathione, and 1-50 units of catalase. PMID- 14555816 TI - Cadmium induced acute lung injury and TUNEL expression of apoptosis in respiratory cells. AB - We examined the ultrastructural features of the lung parenchyma and the expression of apoptosis of the respiratory cells by TUNEL technique. Male Sprague Dawley rats (n=30) were intra-tracheally injected with cadmium (2.5 mg/kg) into both lungs. The light and electron microscopic features of the lung tissues were examined on Days 1, 3, 7 and 10 after the injection of cadmium. Specimen preparations for the light and electron microscopic TUNEL stains were performed. Ultrastructurally, on Days 1 and 3, the alveolar spaces were filled with edematous fluid, and desquamated type I epithelial cells. On Days 7 and 10, the alveolar spaces and interstitium were patchy infiltrated with young fibroblasts and some collagen deposition. The light microscopic TUNEL stain showed that apoptosis of the alveolar cells was most prominent on Day 1, and then the number of apoptosis was markedly decreased on Days 3, 7 and 10. The electron microscopic TUNEL stain showed the electron dense homogenous nuclear expression, and the formation of intra-nuclear blebs which protrude to the outside of nuclei. On Days 7 and 10, there are frequent apoptotic nuclear bodies in the alveolar macrophages. We could examine the identification of the equivocal apoptotic cells and various morphologic expression of apoptotic nuclei on the electron microscopic TUNEL stain. PMID- 14555817 TI - Effects of perfluorocarbon associated high frequency oscillatory ventilation on hemodynamics and gas exchange in the newborn piglets with respiratory distress. AB - We sought to know whether there is a further improvement in gas exchange when partial liquid ventilation (PLV) is added to high-frequency oscillatory ventilation (HFOV) in a piglet model of saline lavage-induced acute lung injury. Seven 7-9 day-old newborn piglets of mixed strain were treated with repeated saline lavage to achieve a uniform degree of acute lung injury. Then, HFOV were applied to the subject. Four animals received two consecutive doses (15 mL/kg) of perfluorodecalin at 30-min interval (PFC+HFOV group). The other three animals remained on HFOV alone (HFOV-only group). Repetitive lung lavage led to a significant acute aggravation in both gas exchange and hemodynamic parameters. Subsequent application of HFOV produced a significant rapid recovery in both gas exchange and hemodynamic parameters to near baseline levels. During and after perfluorodecalin dosing, there were no significant changes in gas exchange or hemodynamic parameters over time in both groups, and no significant differences in gas exchange or hemodynamic parameters between groups. We concluded that the addition of 30 mL/kg of perfluorodecalin to HFOV showed no detrimental effect on hemodynamics, but did not produce a significant improvement in gas exchange over a three-hour period. PMID- 14555818 TI - The clinical significance of antibody screening test including Dia+ panel cell in Asian-Mongoloid populations. AB - The Di(a) antigen is well known as one of the antigens with low incidence among Caucasians; however, it has been discovered with a relatively higher incidence among Mongoloid populations. Thus, it has been speculated that the incidence of unexpected antibody against the Di(a) antigen might be relatively higher among these populations. Hemolytic transfusion reactions (HTRs) and hemolytic disease of the newborns (HDNs) caused by anti-Di(a) have been reported sporadically. However, there has been no prospective study on the incidence of anti-Di(a) in Mongoloid populations particularly. The authors conducted a series of antibody screening tests on 11,219 Korean individuals for 25 months, by using three kinds of screening cells including Di(a) cell. Anti-Di(a) was detected in 8 patients, seven of whom had a history of transfusions or were multigravida. The incidence of anti-Di(a) measured in this study was higher than expected, ranked third among unexpected antibodies identified during the period of the study, so it is strongly recommended that the Di(a+) panel cell must be incorporated into antibody screening test for safer transfusion in Asian-Mongoloid populations. PMID- 14555819 TI - High dose therapy followed by autologous peripheral blood stem cell transplantation as a first line treatment for multiple myeloma: a Korean Multicenter Study. AB - We conducted a phase II multicenter trial to estimate the response and survival of patients with newly diagnosed multiple myeloma to high dose melphalan therapy followed by autologous peripheral blood stem cell transplantation. Eligible patients who had undergone induction with vincristine, adriamycin and dexamethasone (VAD) should have adequate cardiac, pulmonary and renal function (creatinine <2 mg/dL). Melphalan at 200 mg/m2 was used as a conditioning regimen. Eighty patients were enrolled from 13 centers. The median age of the patients was 53 yr (range; 20 to 68 yr). The initial stage was IA/IIA/IIB/IIIA/IIIB in 3/8/1/54/14 patients, respectively. Beta2-microglobulin, CRP and LDH were increased in 74, 42 and 34% of the patients examined. Cytogenetic data were available in 30 patients, and 6 patients showed numeric or structural abnormalities. Two therapy-related mortalities occurred from infection. Among the 78 evaluable patients, CR/PR/MR/NC/PD were achieved in 48/26/2/1/1 patients, respectively. After a median follow-up of 30 months, the median overall and event free survivals were 66 months (95% CI: 20-112) and 24 months (95% CI: 18-29), respectively. This study verifies the efficacy and feasibility of high dose melphalan therapy with autologous stem cell transplantation in newly diagnosed multiple myeloma. PMID- 14555820 TI - Production of stromal cell-derived factor-1 (SDF-1)and expression of CXCR4 in human bone marrow endothelial cells. AB - This study investigated the production of stromal cell-derived factor-1 (SDF-1) and the expression of CXCR4 in human bone marrow endothelial cells (BMECs). Human BMEC cell line BMEC-1 cells expressed SDF-1 mRNA, and conditioned medium induced chemoattraction of CD34+ cells. Migration was not inhibited by pretreating the input cells with pertussis toxin, indicating that the chemoattractive activity was not dependent on SDF-1. Three-day culture of BMEC-1 and primary human BMEC cells produced 1,710+/-204 and 1,050+/-153 pg/mL SDF-1alpha, respectively, which was much less than primary human BM stromal cells (29,536+/-532 pg/ mL). By immuno-histochemistry, CXCR4 was detected in the endothelial cells lining sinusoids, arterioles, and venules in the bone marrow. However, cultured BMECs and BMEC-1 cells did not express CXCR4 on their surfaces. These results indicate that BMECs produce and release small amounts of SDF-1 and express CXCR4 in vivo only. PMID- 14555822 TI - Postoperative outcome in formerly premature infants undergoing herniorrhaphy: comparison of spinal and general anesthesia. AB - To compare the postoperative outcome according to the type of anesthesia, formerly prematured and high-risk infants who had received and weaned ventilator care preoperatively and had undergone inguinal herniorrhaphy were enrolled in this study. Immediate pre- and post-operative respiratory data which contained the lowest respiratory rates, SpO2, heart rates and the incidence of hypoxemia and bradycardia were collected with the incidence of ventilator care, application of continuous positive airway pressure (CPAP), application of oxygen, hospital stay, and respiratory mortality by chart review, retrospectively. Among the twenty-nine infants, fourteen received the general anesthesia (GA group), and fifteen received the spinal anesthesia (SA group). Postoperatively, the infants in the GA group had lower SpO2 (77.1 +/- 20.9% vs. 93.0 +/- 5.5%), higher incidence of hypoxemia (6 vs. 0), ventilator care (5 vs. 0) and application of CPAP (4 vs. 0) than the infants in the SA group. One infant in the GA group died because of acute respiratory failure caused by respiratory syncythial virus pneumonia. We concluded that spinal anesthesia reduces postoperative oxygen desaturation and respiratory morbidity in formerly prematured and high-risk infants who underwent inguinal herniorrhaphy. PMID- 14555821 TI - Serum hepatitis B virus (HBV) DNA levels at different stages of clinical course in patients with chronic HBV infection in an endemic area. AB - The aims of this study were to investigate serum hepatitis B virus (HBV) DNA levels at different clinical stages in patients with chronic HBV infection, and to determine the serum HBV DNA level that discriminated HBeAg-negative chronic hepatitis B(CHB) cases from inactive HBsAg carriers. In all, 222 patients, encompassing 68 HBeAg-positive CHB patients (HBeAg-positive, ALT-elevation), 89 HBeAg-negative CHB patients (HBeAg-negative, ALT-elevation), and 65 inactive HBsAg carriers (HBeAg-negative, ALT-normal), were tested. The ALT levels had been tested more than twice during the previous six months, and the serum HBV DNA levels were quantified by a polymerase chain reaction-based assay. The serum HBV DNA levels of the HBeAg-negative patients were significantly lower than those of the HBeAg-positive patients (median 2.7 x 10(4) vs. 1.6 x 10(8) copies/mL; p=0.000). In addition, the HBV DNA levels of the HBeAg-negative CHB patients were significantly higher than those of the inactive HBsAg carriers (median 2.2 x 10(5) vs. 3.2 x 10(3) copies/ mL; p=0.000). The optimal HBV DNA level for discriminating HBeAg-negative CHB cases from inactive HBsAg carriers was 2.0 x 10(4) copies/mL. The serum HBV DNA levels were lower than the cutoff value in 72.3% (47/65) of the inactive HBsAg carriers, and in 31.5% (28/89) of the HBeAg negative CHB patients. The serum HBV DNA levels differed significantly between these two groups. However, the levels in the two groups overlapped extensively, preventing the definition of a differentiation cut-off value. PMID- 14555823 TI - Serum albumin level correlates with disease severity in patients with Hemorrhagic Fever with Renal Syndrome. AB - Hypoalbuminemia frequently occurs in Hemorrhagic Fever with Renal Syndrome (HFRS), but clinical significance of hypoalbuminemia is not well known. This study was designed to evaluate hypoalbuminemia as a marker of severity of disease in patients with HFRS. We evaluated the relationship between the level of serum albumin and clinical parameters representing the severity of disease in 144 patients with HFRS. The patients were divided into three groups based on the level of serum albumin; Group I (normal serum albumin), Group II (serum albumin <3.5 g/dL and >/=3.0 g/dL), and Group III (serum albumin <3.0 g/dL). Of the total of 144 patients, 42 patients (29.2%) were categorized as Group I, 39 patients (27.1%) as Group II, and 63 patients (43.8%) as Group III. Group III had a higher rate of incidence in episode of hypotension, pulmonary edema than did Group I and Group II. The lowest level of serum albumin was positively correlated with platelet count (r=0.505, p<0.001) and was negatively correlated with leukocyte count (r=-0.329, p<0.001), BUN (r=-0.484, p<0.001), serum creatinine (r=-0.394, p<0.001), and AST (r=-0.251, p=0.002). Our data suggest that hypoalbuminemia frequently occurs in the acute stage of HFRS, and level of serum albumin is associated with the disease severity of HFRS. PMID- 14555825 TI - 99mTc-HDP pinhole SPECT findings of foot reflex sympathetic dystrophy: radiographic and MRI correlation and a speculation about subperiosteal bone resorption. AB - Reflex sympathetic dystrophy (RSD) is a common rheumatic disorder manifesting painful swelling, discoloration, stiffening and atrophy of the skin. Radiographic alterations include small, spotty subperiosteal bone resorption (SBR) and diffuse porosis, and MR imaging shows bone and soft-tissue edema. The purposes of current investigation were to assess 99mTc HDP pinhole SPECT (pSPECT) findings of RSD, to correlate them with those of radiography and MRI and to speculate about causative mechanism of SBR which characterizes RSD. pSPECT was performed in five patients with RSD of the foot. pSPECT showed small, discrete, spotty hot areas in the subperiosteal zones of ankle bones in all five patients. Diffusely increased tracer uptake was seen in the retrocalcaneal surface where the calcaneal tendon inserts in two patients with atrophic RSD. pSPECT and radiographic correlation showed spotty hot areas, that reflect focally activated bone turnover, to closely match with SBR. Further correlation with MRI showed both spotty hot areas and SBR to coincide in location with the insertions of ligaments and tendons, onto which pulling strain is constantly exerted. In contrast, the disuse osteoporosis in unstrained bones did not show any more significantly increased tracer uptake than normal cancellous bones. PMID- 14555824 TI - Inhibitory effects of potassium channel blockers on carbachol-induced contraction in rat detrusor muscle. AB - We present accidental findings that potassium channel blockers, such as tetraethyl-ammonium (TEA) or 4-aminopyridine (4-AP), inhibit the sustained tonic contraction induced by carbachol in rat detrusor muscle strips. The relatively lower concentrations (<2 mM) of TEA and 4-AP inhibited phasic and tonic contractions induced by 5 micro M carbachol, whilst the relatively higher concentrations of TEA and 4-AP (>5 mM) potentiated phasic contractions. The potentiation of phasic contraction was not observed in nicardipine pretreated condition. In nicardipine pretreated condition, the concentration-response curves for the negative inotropic effect of potassium channel blockers were shifted to the right by the increasing concentration of carbachol from 0.5 microM to 5 microM. IC50 was changed significantly from 0.19 to 0.64 mM (TEA) and from 0.21 to 0.96 (4-AP). Such inhibitory effects were also observed in Ca2+ depleted condition, where 0.1 mM EGTA and 1 microM thapsigargin were added into Ca2+ free solution. In conclusion, inhibitory effects of potassium channel blockers on carbachol-induced contraction may be ascribed to the direct inhibition of receptor-agonist binding. PMID- 14555826 TI - Distribution of lengths of the normal femur and tibia in Korean children from three to sixteen years of age. AB - To develop a standard growth curve of the lower extremity in Korean children from 3 to 16 yr of age, the lengths from a total of 2087 normal long bone segments (582 femurs and 645 tibias in boys, and 417 femurs and 443 tibias in girls) were measured. Children were grouped by years of bone age, which was determined by using the Korean specific bone age standard; TW2-20 method. The growth spurt occurred in girls from eight to eleven years by bone age, and in boys from eleven to thirteen years. The mean tibial length relative to the mean femoral length was 0.78 in boys and 0.79 in girls. The overall growth pattern was similar to that observed in American children in the 1960s. Korean children and adolescents appear to have a different tempo of skeletal maturation during pubertal growth from that of English and American children and adolescents. The Korean standard growth curve and the Korean bone age chart allow determination of the presence of any existent growth abnormalities and prediction of future remaining growth in lower extremities. These normative growth standards can be used for leg-length equalization purposes in children with anisomelia. PMID- 14555827 TI - Biochemical characteristics of a Korean patient with mucolipidosis III (pseudo Hurler polydystrophy). AB - We performed a biochemical study on the patient with mucolipidosis III (ML-III, pseudo-Hurler polydystrophy) in Korea. Confluent fibroblasts from the patient and from normal controls were cultured for 4, 12, 24, 48, and 72 hr, respectively. Lysosomal enzyme activities in culture media after different incubation times and in plasma, leuko-cytes, and fibroblasts were determined. Most of the leukocyte lysosomal enzymes were within normal limits or slightly lowered; however, plasma lysosomal enzyme activities such as those of hexosaminidase and arylsulfatase A were markedly increased. Numerous phase-dense inclusions were present in the cytoplasm of cultured fibroblasts. Lysosomal enzyme activities of fibroblasts were markedly decreased except for beta-glucosidase. The rates of increase of the lysosomal enzyme activities with incubation time were greater in the culture medium of the patient than in normal control, whereas no difference in the beta glucosidase activity of the culture media of the patient and the control was found. This study describes the first case of ML-III in Korea, with its typical biochemical characteristics, i.e., a problem with targeting and transporting of lysosomal enzymes which results in a marked increase in plasma lysosomal enzyme activities and a high ratio of extracellular to intracellular lysosomal enzyme activities in cultured fibroblasts. PMID- 14555828 TI - Rapid detection of duplication/deletion of the PMP22 gene in patients with Charcot-Marie-Tooth disease Type 1A and hereditary neuropathy with liability to pressure palsy by real-time quantitative PCR using SYBR Green I dye. AB - Mutations and altered gene dosage of the peripheral myelin protein (PMP22) gene in chromosome 17p11.2-12 are the main causes for hereditary neuropathies, accounting for approximately 70% of all cases. Patients with duplication of the PMP22 develop Charcot-Marie-Tooth disease type 1A (CMT1A) and deletion of one PMP22 allele leads to hereditary neuropathy with liability to pressure palsy (HNPP). Twenty patients with CMT1A, 17 patients with HNPP, and 18 normal family members and 28 normal controls were studied by real-time quantitative PCR using SYBR Green I on the ABI 7700 Sequence Detection System. The copy number of the PMP22 gene was determined by the comparative threshold cycle method and the albumin was used as a reference gene. The PMP22 duplication ratio ranged from 1.45 to 2.06 and the PMP22 deletion ratio ranged from 0.42 to 0.64. The PMP22 ratio in normal controls, including normal family members, ranged from 0.85 to 1.26. No overlap was found between patients with CMT1A or patients with HNPP and normal controls. This method is fast, highly sensitive, specific, and reproducible in detecting PMP22 duplication and deletion in CMT1A and HNPP patients, respectively. PMID- 14555829 TI - Initial dose cascade of TTS fentanyl with proper adjuvant medications in cancer pain. AB - According to the three step-ladder analgesics in patients with cancer pain, adjuvant drugs are required for pain relief according to the pain character and also to reduce side effects of opioids. Pain clinicians sometimes want to decide to jump directly from naive and mild opioid to transdermal therapeutic system (TTS) fentanyl with less side effects. We investigated the safety, efficacy, and satisfaction of the patients of TTS fentanyl converting from opioid-naive and mild-opioid with adjuvant drug medications in related to dose cascade of TTS fentanyl. Both opioid-naive (n=3) and opioid-using (n=34) patients started with TTS fentanyl in the lowest available delivery rate (25 microg/hr) with rescue medication. A numeric rating scale (NRS, from 0=no pain to 10=worst pain imaginable), satisfaction of the patients with the transdermal therapy and side effects were recorded everyday during 29 days. Average reductions of NRS scores were 1.79 and 2.77, and the mean doses were 35.14 and 44.12 microg/hr on the 15th and 29th day, respectively. Reported level of satisfaction with the transdermal patch and generalized pain management were 'completely satisfied' and 'satisfied'. Frequent side effects were nausea, vomiting, and constipation. In conclusion, initial application of TTS fentanyl with proper adjuvant medications is effective, safe, and well tolerated. PMID- 14555830 TI - A case of human cyclosporiasis causing traveler's diarrhea after visiting Indonesia. AB - This is the first case of human cyclosporiasis reported in Korea. We detected the oocyst of Cyclospora cayetanensis from a 14-yr old girl who complained of persistent diarrhea after traveling to Indonesia. Round oocysts sized about 8 to 9 m with wrinkle on the wall were found in modified acid fast stained stool specimen. Stainability was variable from red to pale. Oocyst wall showed typical autofluorescence under ultraviolet illumination. The exact diagnosis for the cause of diarrhea and treatment for this patient were not provided at the right moment from the hospital since the diagnostic system for the Cyclospora infection was not ready in the clinical laboratory of the hospital. More attention should be paid on Cyclospora as a cause of diarrhea especially for those returning from a trip to the tropics and an adequate diagnostic system for the Cyclospora infection should be implemented in clinical laboratories as soon as possible. PMID- 14555831 TI - A case of primary pericardial undifferentiated sarcoma. AB - Primary pericardial sarcomas are extremely rare. The authors report a case of a 46-yr-old woman in whom a large mediastinal mass was discovered. The patient presented with cough, dyspnea, and orthopnea. Diagnostic investigations, such as echocardiography, computed tomography, and exploratory thoracotomy provided the evidence of a large mass in the mediastinum, attached by a broad base to the superior portion of the pericardium. A excisional biopsy was performed, and histologic examination of a biopsy specimen showed undifferentiated sarcoma. However, the complete removal of the mass was impossible due to adhesion to the adjacent great vessels. After the completion of the chemotherapy the patient was completely asymptomatic. However, follow-up transesophageal echocardiography showed a residual 3 x 4 cm-sized mass. The patient received the radiotherapy with a total dose of 55 Gy over 6 weeks. At present, there is no evidence of disease progression. PMID- 14555832 TI - Telangiectatic focal nodular hyperplasia of the liver: a case detected at birth. AB - A case of telangiectatic focal nodular hyperplasia (FNH) was detected at birth and was surgically removed. Grossly, the lesion was a solitary nodule and showed vague nodularity, appearing as an adenoma-like mass with fine fibrous septa, but having no macroscopic scar. On microscopic scale, the mass typically had neither fibrous central scar nor hyperplastic nodules different from the usual FNHs. The hepatic plates were separated by sinusoidal dilatation, sometimes alternating with areas of marked ectasia. Instead of large fibrous scar, thin fibrous septa were often found, and contained abnormal tortuous large arteries. These high pressure vessels were connected directly into the adjacent sinusoids and made marked dilation of sinusoids. Bile ductular proliferation was also noted in the thin fibrous septa. To our knowledge, this is considered to be the first reported case of telangiectatic FNH detected at birth. PMID- 14555833 TI - A spontaneously ruptured gastric stromal tumor with cystic degeneration presenting as hemoperitoneum: a case report. AB - We report a case of a 38-yr-old man with a spontaneously ruptured gastric stromal tumor presenting as hemoperitoneum in outpatient clinic. He visited our hospital with generalized abdominal pain after abdominal CT scan for the evaluation of the asymptomatic palpable abdominal mass. Repeated abdominal CT scan showed a size decrement of cystic mass compared with the previous abdominal CT scan, and newly developed fluid collection in the left paracolic gutter. An emergency laparotomy revealed a ruptured gastric stromal tumor with bloody fluid in the peritoneal cavity. Immunohistochemical examination revealed positive reactivity to C-kit protein and CD34. The patient presented with hemoperitoneum due to spontaneous rupture of the tumor, which is an extremely rare complication. PMID- 14555834 TI - Retroperitoneal abscess complicated by acupuncture: case report. AB - With acupuncture treatment becoming an increasingly popular analgesic, there have been increasing reports on its associated complications. Although pneumothorax is the most frequently reported injury caused by acupuncture needles, infectious complications may not be uncommon. Most infectious complications show less serious clinical manifestations than pneumothorax, but retroperitoneal or intraabdominal abscess caused by acupuncture may be much more serious conditions. We experienced a 56-yr-old male diabetic patient presenting with serious retroperitoneal abscess after acupuncture treatments. Emergency operative drainage with adequate antibiotic therapy was performed. Bacterial culture of blood and closed pus specimens recovered Klebsiella pneumoniae. In addition to application of better knowledge on anatomy, appropriate antiseptic practice by practitioners will reduce many serious complications associated with acupuncture. PMID- 14555835 TI - Emphysematous prostatic abscess due to Klebsiella pneumoniae: report of a case and review of the literature. AB - Emphysematous prostatic abscess is a very rare form of prostatitis. Emphysematous prostatic abscess due to Klebsiella pneumoniae may have a poor prognosis according to a few previous reports. We report a rare case of successfully treated emphysematous prostatic abscess with cystitis due to Klebsiella pneumoniae in a 50-yr-old man with 15-yr history of diabetes mellitus. The patient was referred to the emergency room of our hospital. The KUB film revealed gas shadows in the lower pelvic area suggestive of emphysematous cystitis or emphysematous prostatic abscess. The gas was mainly occupying the prostate and was also seen in the bladder on pelvic CT. The patient was successfully treated with long-term antibiotic use and additional percutaneous drainage of the abscess. Emphysematous prostatic abscess may be misdiagnosed as emphysematous cystitis due to the similar location of gas shadows on radiography. Computerized tomography and transrectal ultrasonography are helpful in making the diagnosis of emphysematous prostatic abscess. Appropriate use of effective antibiotics with drainage of pus is the best treatment. This case emphasizes the importance of timely and accurate diagnosis followed by appropriate treatment in emphysematous prostatic abscess in diabetic patients. PMID- 14555836 TI - A case of hypertrophic osteoarthropathy associated with nasopharyngeal carcinoma in a child. AB - Hypertrophic osteoarthropathy is characterized by clubbing of the digital tips and periosteal reaction of long bones. Most of the cases are associated with malignancy or other conditions such as congenital heart disease, liver cirrhosis, pulmonary fibrosis, biliary atresia, and gastrointestinal polyps. Hypertrophic osteoarthropathy associated with malignancy is rare in children. A few cases of hypertrophic osteoarthropathy in children with nasopharyngeal carcinoma have been reported, however, there has been no report of such case in Korea. We present a case of hypertrophic osteoarthropathy associated with nasopharyngeal carcinoma with lung metastasis in a 14-yr-old boy. In this case, hypertrophic osteoarthropathy regressed after intensive chemotherapy, but subsequently the patient died of progressive lung metastasis. PMID- 14555837 TI - Warfarin-associated fetal intracranial hemorrhage: a case report. AB - A 27-yr-old woman who had been taking warfarin for 10 yr after mitral valve replacement became pregnant. After knowing her pregnancy, she received heparinization for nine weeks instead of warfarin, and took oral anticoagulant again. At 24 weeks of gestation, fetal ultrasound and MRI showed a left subdural hematoma, and the pregnancy was terminated. Subdural hematoma was demonstrated on autopsy. Fatal bleeding of the fetus is a rare complication of maternal warfarin medication, occurring mostly in the second or third trimester. There is no alternative regimen available, so that regular monitoring by fetal ultrasound and strict control of warfarin dose with regular measurement of prothrombin time are the best way to prevent intrauterine fetal death due to bleeding. PMID- 14555838 TI - Chordoid meningioma: a case report. AB - The term "chordoid meningioma" means meningioma, which is pathologically similar to chordoma, and previously reported that rarely associated with microcytic anemia and/or dysgammaglobulinemia especially in pediatric population. We present a case of this rare variant, which comprises less than 0.5% of all meningiomas. A 33-yr-old man visited our hospital, complaining visual field defect worsening over 7 yr. Neurological examination showed left homonymous hemianopsia. The brain magnetic resonance imaging revealed well enhancing right temporo-occipital mass with cystic portion. Histopathologic findings of resected tumor were compatible with chordoid meningioma which included trabeculae of eosinophilic, vacuolated cells in a myxoid matrix with prominent lymphoplasmacellular infiltration. The neoplastic cells were positive for vimentin and epithelial membrane antigen and negative for glial fibrillary acidic protein and cytokeratin. This is an adult case of chordoid meningioma without anemia or dysgammaglobulinemia. PMID- 14555839 TI - The roles of soluble osteopontin using osteopontin-transgenic mice in vivo: proliferation of CD4+ T lymphocytes and the enhancement of cell-mediated immune responses. AB - We generated transgenic mice expressing osteopontin (OPN) under the control of the alpha(1)-antitrypsin promoter. These mice (OPN-T mice) expressed OPN mRNA in liver and kidney, and released a large amount of plasma OPN, which increased after stimulation with turpentine oil. Before sensitization, the number of CD4+ T cells in lymph nodes was significantly higher in OPN-T than nontransgenic mice, and that in spleen was slightly higher, whereas that of CD8+ T cells was no different between OPN-T and nontransgenic mice. After sensitization, the CD4+ T cell numbers in spleen increased significantly, while there were almost no changes in the CD8+ T cells in lymph nodes and spleen. The intensity of contact hypersensitivity responses to 2,4-dinitrofluorobenzene (DNFB) was obviously enhanced in OPN-T mice. In the delayed-type hypersensitivity (DTH) model elicited by DNFB, the number of CD8+ T cells among DNFB-2,4,6-trinitrobenzenesulfonic acid (TNBS)-peritoneal exudate cells was significantly higher in OPN-T than nontransgenic mice, while there was almost no difference in that of CD4+ T cells. Adoptive transfer experiments revealed that the enhanced reactivity is carried by CD4+ and CD8+ T cells, respectively, although the ability of transferring DTH was significantly lower in CD8+ than in CD4+ T cells. The enhancement of CD8+ T cell migration was observed in OPN-T mice. These results suggest that OPN induces a proliferation of effector CD4+ and CD8+ cells in cell-mediated reactions and plays a role in the migration of CD8+ T cells. PMID- 14555840 TI - Expression of uridine diphosphate N-acetyl-alpha-D-galactosamine: polypeptide N acetylgalactosaminyl transferase 3 in adenocarcinoma of the pancreas. AB - Uridine diphosphate (UDP) N-acetyl-alpha-D-galactosamine:polypeptide N acetylgalactosaminyl transferase 3 (GalNAc-T3), one of the enzymes that catalyze the initial glycosylation of mucin type O-linked proteins, was shown to associate with the differentiation of cancer cell lines and the prognosis of some kinds of cancers. In the present study, the association of GalNAc-T3 expression with clinicopathologic features of pancreatic adenocarcinoma and patients' survival was examined. The level of expression of GalNAc-T3 was analyzed immunohistochemically in paraffin-embedded tumor samples from surgically resected specimens of 59 patients with pancreatic ductal adenocarcinoma. The correlations of GalNAc-T3 expression with clinicopathologic features and prognosis were studied. Thirty-five tumors showed high- intensity GalNAc-T3 staining, whereas 24 showed low-intensity staining. A close association was observed between GalNAc-T3 staining intensity and histologic differentiation and the stage of the tumors. The low-intensity group showed a high rate of the poorly differentiated subtype (p < 0.001) and T4 of the pTNM staging system (p < 0.05). These findings indicate that the expression of GalNAc-T3 is associated with the differentiation and aggressiveness of ductal adenocarcinoma of the pancreas. PMID- 14555841 TI - Expression of RHOC is associated with metastasis of gastric carcinomas. AB - OBJECTIVES: RHOC, a member of the RAS-related small GTPase protein family, regulates cytoskeletal structures and has the potential to transform cultured cells. It has recently been reported that RHOC contributes to the metastatic phenotype of melanoma cells. The purpose of this study was to clarify its biological relevance to gastric carcinogenesis and metastasis. METHODS: We examined the expression of RHOC by quantitative RT-PCR in 51 cases of gastric carcinoma tissues from prior surgical cases (intestinal type: 24 cases, diffuse type: 27 cases) and in 8 gastric carcinoma cell lines. RESULTS: RHOC expression levels in primary tumors were significantly higher in cases with metastasis than in those without metastasis (p = 0.0202; Mann-Whitney U test). RHOC expression levels in primary tumor and their metastatic tumors were significantly higher than their corresponding nonneoplastic mucosa (p = 0.0357, and 0.0173, respectively; Wilcoxon signed rank test). RHOC mRNA expression was confirmed in the gastric carcinoma cell lines. CONCLUSION: Our findings suggest that elevated expression of the RHOC gene may be involved in the metastasis of gastric carcinomas and may be a good genetic marker for the prediction of a metastatic potential. PMID- 14555842 TI - Association between expression levels of CA 19-9 and N-acetylglucosamine-beta;1,3 galactosyltransferase 5 gene in human pancreatic cancer tissue. AB - OBJECTIVE: CA 19-9, equivalent to Sialyl Lewis antigen, is a well-known tumor marker in pancreatic cancer. At the initial step of the biosynthesis of CA 19-9, N-acetylglucosamine-beta1,3-galactosyltransferase (beta3Gal-T) transfers galactose to N-acetylglucosamine (GlcNAc). Recently, beta3Gal-T5 has been presumed to be related to the formation of the type 1 chain in an in vitro experiment in terms of kinetic enzyme characterization. The purpose of this study was to investigate which beta3Gal-T is related to the synthesis of CA 19-9 in human pancreatic cancer tissues. METHODS: We examined beta;3Gal-T1, T2, T3, T4, and beta;3Gal-T5 mRNA expressions in 13 noncancerous and cancerous tissues of the human pancreas using real-time polymerase chain reaction, and compared those gene expression levels with the immunoreactivity of CA 19-9 and its precursor DUPAN-2 in cancerous tissues. RESULTS: Beta;3Gal-T5 gene expression significantly augmented in cancerous tissues, when compared with the adjacent noncancerous tissues. Additionally, there was a good correlation between BETA;3GAL-T5 gene transcription levels and immunohistochemical grades of CA 19-9 or its precursor DUPAN-2 in cancerous tissues. However, no correlation was observed between beta;3Gal-T1, T2, T3, and beta;3Gal-T4 gene expression levels and CA 19-9 or DUPAN-2 immunoreactive grades in cancerous tissue. CONCLUSION: beta3Gal-T5 is presumed to be responsible for the synthesis of CA 19-9 in pancreatic cancer tissue. PMID- 14555843 TI - Inflammatory cytokines, pleuropneumonia infection and the effect of dexamethasone. AB - OBJECTIVES: Actinobacillus pleuropneumoniae causes an often fatal infection of swine due to pleuropneumonia. To determine if inflammatory cytokines are associated with A. pleuropneumoniae-induced pneumonia, infected and noninfected animals were concomitantly administered saline or dexamethasone. METHODS: Twenty four swine were treated with saline, A. pleuropneumoniae, dexamethasone, or A. pleuropneumoniae and dexamethasone (n = 6). The plasma levels of TNF-alpha, IL 1beta, IL-6, IL-8, and IL-10 were examined through time of necropsy (72 h). Gross pathology and histopathology was performed on all animals. RESULTS: Dexamethasone had no effect on A. pleuropneumoniae-induced increases in lung/body weight ratios. Gross pathology of the infected pigs included fibrinous pleuropneumonia with necrosis and hemorrhage in a focal to a multifocal pattern. Histopathology of infected pig lungs revealed necrotizing extensive, fibrinopurulent pneumonia with edema and fibrinopurulent pleuritis. Plasma IL-6 levels were elevated in A. pleuropneumoniae-infected animals beginning 6 h after infection. Dexamethasone treatment did not alter A. pleuropneumoniae-induced plasma IL-6 levels. A. pleuropneumoniae infection did not elicit plasma levels of TNF-alpha, IL-1beta, IL-8, or IL-10. CONCLUSION: These results suggest that the pneumonia caused by A. pleuropneumoniae infection is not due to the release of systemic inflammatory cytokines. PMID- 14555844 TI - Altered expression of ARPP protein in skeletal muscles of patients with muscular dystrophy, congenital myopathy and spinal muscular atrophy. AB - OBJECTIVES: Ankyrin-repeated protein with PEST and a proline-rich region (ARPP) is a recently identified protein with 4 ankyrin-repeated motifs in its central portion. Type 1 myofibers of skeletal muscle express high levels of ARPP. Recently, we have found that ARPP expression was induced in mouse denervated skeletal muscle. This led us to hypothesize that ARPP expression might be induced in skeletal muscle under some pathological conditions. METHODS: In this study, we performed immunohistochemical analysis of ARPP expression in biopsy specimens of muscle tissue from 15 patients with muscular dystrophies (MDs), 13 with congenital myopathies and 11 with spinal muscular atrophies (SMAs). RESULTS: The ARPP expression levels of all the specimens from MD patients appeared to be lower than control muscle levels. In contrast, the specimens from the 13 patients with congenital myopathies were all ARPP positive. We also found increased numbers of ARPP-positive myofibers in patients with congenital myopathies, and these myofibers co-expressed the slow myosin heavy chain. Indeed, it has been reported that type 1 myofibers are predominant in patients with congenital myopathies, suggesting that increased numbers of ARPP-positive myofibers in such patients may be associated with increased numbers of type 1 fibers. In patients with SMAs, we found that ARPP-positive myofibers tended to be distributed in groups. As grouped myofibers have been reported to result from the process of denervation, innervation and subsequent denervation of re-innervated myofibers, the grouped ARPP-positive myofibers in SMA patients may result from denervation of the motor units. CONCLUSIONS: These findings suggest that evaluation of ARPP may be helpful for the histological diagnosis of muscle diseases. PMID- 14555845 TI - Differential expression of protease-activated receptors 1, 2, and 4 on human endothelial cells from different vascular sites. AB - OBJECTIVE: Protease-activated receptors (PARs) mediate DNA synthesis in endothelial cells when activated by serine proteases. However, despite the existence of heterogeneity among endothelial cells from each tissue, the responses to PAR-1, PAR-2, and PAR-4 activation are poorly defined and compared between endothelial cells from different sites. The aim of this study was to investigate whether PAR-mediated DNA synthesis differed in various endothelial cell types. METHODS: We examined the incorporation of BrdU by human pulmonary artery endothelial cells (HPAECs), human aortic endothelial cells (HAECs), and human umbilical vein endothelial cells (HUVECs). RESULTS: When the endothelial cells were treated with the selective PAR-1-activating peptide, SFLLRN, HAECs showed the highest BrdU incorporation rate (182 +/- 28%). In contrast, treatment with the PAR-2-activating peptide, SLIGKV, resulted in the highest BrdU incorporation rate (173 +/- 37%) in HPAECs, when pretreated with TNF-alpha. The PAR-4-activating peptide, GYPGQV, induced DNA synthesis in HPAECs and HAECs, but not in HUVECs. CONCLUSION: These findings suggest that each PAR preferentially targets an endothelial cell type, and thus plays a distinct role in diverse physiological or pathological conditions. PMID- 14555846 TI - Human herpesvirus-6: a short review of its biological behavior. AB - HHV-6 shows a widespread distribution with life-long persistence. The virus is frequently reactivated, yet remains clinically inapparent unless the patient is immunodeficient in some way. Even then, HHV-6 reactivation may simply enhance the pathogenicity of other viruses or existing autoimmune disorders rather than becoming a pathogen itself. Future clinical studies need to focus on such indirect viral influences mediated through molecular mimicry and interference with cell receptor expression, and cytokine and chemokine network regulation. Nevertheless, such disturbances may afford therapeutic intervention to disrupt herpesvirus interference and improve certain disease processes. There are only a few diseases for which an immediate causal relationship to HHV-6 infection has been suggested. PMID- 14555847 TI - Safety evaluation of GX-12, a new HIV therapeutic vaccine: investigation of integration into the host genome and expression in the reproductive organs. AB - AIDS is one of the greatest infectious disease threats to human health despite the extensive efforts made since the discovery of HIV in 1983. The use of plasmid DNA vaccination to elicit humoral and cell-mediated immune responses against HIV infection has produced promising results in animal and in human trials. However, there are several safety concerns about the use of a DNA vaccine, which include the possibility of integration into the host genome, adverse immunopathology, and anti-DNA autoantibody induction. In this study, we examined the potential integration and distribution of GX-12, a new therapeutic vaccine for HIV infection, at various times in muscles and reproductive organs of rats. Animals of both sexes were injected with GX-12 at the dose of 400 microg/animal i.m. once a week for 4 weeks, and host genome integration and tissue distribution were examined on day 1, 5, 15, 30 and 45 days after the final injection. A PCR-based assay revealed that GX-12 was not integrated into the host genome, nor expressed in reproductive organs at any time. These findings suggest that the risk of mutation or germline transmission due to GX-12 injection is negligible. PMID- 14555848 TI - Analysis of T cell receptor variable regions and complementarity determining region 3 of infiltrating T lymphocytes in the liver of patients with chronic type B hepatitis. AB - OBJECTIVES: T cell receptor (TCR) variable regions and complementarity determining regions 3 (CDR3) of infiltrating T cells in the livers of patients with chronic hepatitis B were examined in detail. METHODS: TCR usage was examined by the inverse polymerase chain reaction in 2 hepatitis B e antigen-positive patients, one at an early stage and the other at an advanced stage. RESULTS: In 61 productively rearranged clones bearing TCRalpha genes in the liver of the patient with early stage chronic hepatitis B, the Valpha 7.2 gene segment was most frequently used (16.4%), while the other Valpha gene segments were used less frequently. All but one of the clones with the Valpha7.2 gene were joined with the Jalpha33 gene, and the CDR3 regions of these clones were the same length and similar in amino acid sequence. In contrast, in 68 productively rearranged clones from the liver of the patient with advanced stage disease, all Valpha gene segments were used less frequently, and combinations of the Valpha and Jalpha gene segments varied. CONCLUSIONS: Our data suggest a relatively restricted usage of TCR in the liver in early stage chronic hepatitis B. PMID- 14555849 TI - E/NS1 modifications of dengue 2 virus after serial passages in mammalian and/or mosquito cells. AB - OBJECTIVE: Dengue viruses are routinely maintained in nature by transmission cycles involving the passage of virus between humans and AEDES mosquitoes. The number of dengue virus lineages has been increasing over time. The aim of this study was to identify the genetic diversity of dengue 2 virus serially transferred in mammalian and/or mosquito cells. METHODS: The E/NS1 gene of dengue 2 virus variants derived from serial passages in Vero or C6/36 cells, or alternately in both cell systems, was amplified and sequenced in order to observe gene modification after serial passages. RESULTS: Three nucleotides (two in E and one in NS1) or two amino acids (one each in E and NS1) changed in the virus that was continuously cultured in Vero cells for 20 passages, whereas four nucleotides (two each in E and NS1) or three amino acids (one in E and two in NS1) changed in the virus cultured for 30 passages. The genome of dengue 2 virus remained stable even when the virus was serially transferred in C6/36 cells for 30 generations. However, there was one amino acid substitution (E46 I-->V) resulting from a single nucleotide change in the E region of dengue 2 virus alternately transferred in C6/36 and Vero cells for either 20 or 30 passages. In addition, dengue 2 virus obtained from serially cultured Vero cells usually replicated better when it reinfected Vero cells, reflecting its high adaptation fitness to the host cell. CONCLUSIONS: It is concluded that genetic changes of dengue 2 virus are constrained in Vero (mammalian) cells, resulting in a variety of genome related quasispecies populations. Some populations of the virus are subsequently selected by and genetically (at least in the E/NS1 portion of the viral genome) maintained in C6/36 (mosquito) cells during replicative competition. PMID- 14555851 TI - Full-length sequences of six hepatitis E virus isolates of genotypes III and IV from patients with sporadic acute or fulminant hepatitis in Japan. AB - OBJECTIVE: Ranges of variation and conservation in sequence need to be defined for detecting and genotyping hepatitis E virus (HEV). METHODS: Six HEV isolates from Japanese patients were sequenced over the entire genome and compared phylogenetically along with 16 reported HEV isolates, including two from pigs. RESULTS: Three of the six HEV isolates were of genotype III, and the remaining three were of genotype IV. Local clusterings of Japanese HEV isolates were observed in the phylogenetic analyses, including a swine HEV isolate reported previously (swJ570). All six HEV isolates possessed three open reading frames (ORFs). The ORF3 in the three isolates of genotype III were in a different reading frame, while that in the three isolates of genotype IV were in the same reading frame as ORF1. A stretch of 46-96 nucleotides was identified, point mutations and deletions in which were specific for the four genotypes (I-IV). A polymerase chain reaction method was developed with 9 nested universal primers, deduced from conserved regions in the 5'-terminal sequences of the 22 HEV genomes. CONCLUSIONS: Conserved and genotype-specific variation in HEV sequences, identified in the comparison of 22 full-length genomes, would be useful in designing primers for sensitive detection and specific genotyping of HEV RNA. PMID- 14555850 TI - Long-term outcome, with monitoring of platelet counts, in patients with chronic hepatitis C and liver cirrhosis after interferon therapy. AB - OBJECTIVE: Because the determination of the stage of fibrosis depends on rather subjective judgment, more objective parameters are needed. In this study, we followed the long-term outcome, with monitoring of platelet counts, in patients with chronic hepatitis C or liver cirrhosis (LC) who had undergone interferon (IFN) therapy. METHODS: 596 patients who were diagnosed at our institute from 1987 to 1998 with chronic hepatitis C and LC were treated with IFNs. A further 58 patients were not treated (NT). The annual rate of changes in platelet counts were calculated and compared for IFN-treated and NT patients. RESULTS: The relationship between the efficacy of IFN therapy and the incidence of hepatocellular carcinoma (HCC) showed that the patients who were virologic sustained responders (VSR) had a significantly lower incidence of HCC than the nonresponders (NR) and NT patients. The change in platelet counts was +4,350/microl/year in the VSR, +1,010/microl/year in the biochemical sustained responders (BSR), -4,540/microl/year in the NR and -6,180/microl/year in the NT patients, indicating a significant platelet increase in the VSR, a decrease of the same magnitude in the NR and NT patients, and no change in the BSR. The cumulative probability of developing HCC and liver failure was significantly higher in groups with decreased platelet counts than in groups with increased platelet counts among patients who had undergone IFN therapy. Multivariate analyses revealed that a decrease in platelet counts was the cardinal risk factor for development of HCC and liver failure in chronic hepatitis C or LC patients. CONCLUSION: Investigation of platelet counts was useful for determining the long term outcome of patients who had undergone IFN therapy and for predicting the development of HCC. PMID- 14555852 TI - Characterization of HeLa cells persistently infected with influenza virus B/Lee/40 with respect to telomerase activity and apoptosis. AB - OBJECTIVE: The purpose of this study was to examine telomerase activity and apoptotic changes in HeLa cells persistently infected with influenza viruses B/Lee/40 (He/Le cells). METHODS: He/Le cells were established as described previously [Intervirology 2002;45:67-70], and passaged twice a week. The existence of influenza virus genes was monitored by the reverse transcription polymerase chain reaction (RT-PCR). Telomerase activities in He/Le cells were assayed by Telochaser (stretch PCR method). Apoptotic changes in He/Le cells were examined using the Apoptotic DNA Ladder Kit and the In situ Cell Death Detection Kit, Fluorescence. RESULTS: In He/Le cells, (1) all eight influenza virus genes were detected by RT-PCR until 62 days post infection (p.i.); (2) only nucleoprotein gene remained detectable until 120 days p.i.; (3) telomerase activity of He/Le cells normalized to those of uninfected HeLa cells was remarkably decreased (16-55% of control) during the persistence of influenza and recovered up to 80% of control on day 168 p.i. when no influenza virus gene was detected by RT-PCR, and (4) no apoptotic changes were detected despite the continuous existence of influenza virus genes. CONCLUSION: In He/Le cells, telomerase activity was suppressed exclusively during the persistence of influenza, and no apoptotic changes were detected. PMID- 14555853 TI - Focus on cat allergen (Fel d 1): immunological and aerodynamic characteristics, modality of airway sensitization and avoidance strategies. AB - The increasing frequency of pet ownership (especially cats) in many industrialized countries has raised the level of exposure to the allergens produced by these animals. Moreover, it is likely that modern energy-saving systems and the wide use of upholstered furniture has resulted in closer contact between cats (and their allergens) and humans. Many different methods have been developed to quantify the main cat allergen (Fel d 1) in settled dust and in ambient air. The threshold levels of cat allergen inducing sensitization or triggering respiratory symptoms in sensitized patients have been calculated in settled dust, but airborne amounts of Fel d 1 probably represent a more reliable index of allergen exposure. Noticeably, the amount of Fel d 1 may be relatively high also in confined environments where cats have never been kept. It has been demonstrated that clothes of cat owners are the main source for dispersal of allergens in cat-free environments. This fact may be of relevance, because recent studies have shown that allergic sensitization to cats is more likely to develop in children exposed to moderate levels of this allergen than in children exposed to high amounts of Fel d 1. The ubiquity of cat allergen may justify the common observation that allergen avoidance is often insufficient to reduce the risk of developing allergic sensitization and/or symptom exacerbation in highly susceptible patients. Further efforts are needed to improve the efficacy of Fel d 1 avoidance strategies to try to reduce the risk of allergic sensitization to this allergen. PMID- 14555854 TI - Renaissance of the blocking antibody concept in type I allergy. AB - Formation of IgE antibodies against per se harmless antigens (i.e. allergens) is the hallmark and key pathomechanism of type I allergy, a hypersensitivity disease affecting more than 25% of the population. Classical experiments performed more than 65 years ago demonstrated that allergen-specific IgG antibodies, termed blocking antibodies, can antagonize the cascade of allergic inflammation resulting from allergen recognition by IgE antibodies. However, controversial results have questioned the protective role of IgG antibodies in allergic diseases. Here, we review recent data demonstrating that blocking antibodies inhibit allergen-induced release of inflammatory mediators from basophils and mast cells as well as IgE-facilitated allergen presentation to T cells, thus leading to suppression of T cell activation. Furthermore, it has been reported that the development of blocking antibodies is associated with reduced boosts of allergen-specific IgE production in patients receiving allergen-specific immunotherapy. These findings suggest that blocking antibodies have protective activity by inhibiting immediate as well as late inflammatory responses and long term ameliorating activity on the allergic immune response by antagonizing the underlying IgE production. Induction of blocking antibodies is thus an important mechanism underlying allergen-specific immunotherapy. In addition, passive administration of blocking antibodies may be considered as a potential therapeutic strategy for allergic diseases. PMID- 14555855 TI - Three-dimensional structure of Fel d 1, the major allergen in cat. PMID- 14555857 TI - Interleukin-10 influences the expression of MRP8 and MRP14 in human dendritic cells. AB - BACKGROUND: We investigated the differential gene expression of human dendritic cells (DC) under the influence of the cytokine interleukin-10 (IL-10). METHODS: The gene expression profiles of IL-10-treated DC and untreated DC (untreated control) were compared using cDNA expression array. RESULTS: In this array, several differentially expressed transcripts could be identified. Two of them, the transcripts for the myeloid-related proteins (MRPs) MRP8 and MRP14, were examined more closely. The transcripts encoding for these proteins were expressed in mature untreated DC, and significantly induced in IL-10-treated DC. CONCLUSIONS: These data demonstrate that the genes for MRP8 and MRP14 are expressed in various DC subpopulations in different intensity, depending on their maturational status and are increased after the treatment with IL-10. Expression of the genes for MRP8 and MRP14 in DC has not been reported before. These results point to different, still unknown functions of these two proteins in DC: to a role in DC maturation, and possibly to an influence on the creation of tolerogenic T cells under the influence of IL-10. PMID- 14555856 TI - Ana o 2, a major cashew (Anacardium occidentale L.) nut allergen of the legumin family. AB - BACKGROUND: We recently cloned and described a vicilin and showed it to be a major cashew allergen. Additional IgE-reactive cashew peptides of the legumin group and 2S albumin families have also been reported. Here, we attempt to clone, express and characterize a second major cashew allergen. METHODS: A cashew cDNA library was screened with human IgE and rabbit IgG anti-cashew extract antisera, and a reactive nonvicilin clone was sequenced and expressed as a fusion protein in Escherichia coli. Immunoblotting was used to screen for reactivity with patients' sera, and inhibition of immunoblotting was used to identify the corresponding native peptides in cashew nut extract. The identified allergen was subjected to linear epitope mapping using SPOTs solid-phase synthetic peptide technology. RESULTS: Sequence analysis showed the selected clone, designated Ana o 2, to encode for a member of the legumin family (an 11S globulin) of seed storage proteins. By IgE immunoblotting, 13 of 21 sera (62%) from cashew-allergic patients were reactive. Immunoblot inhibition data showed that the native Ana o 2 constitutes a major band at approximately 33 kD and a minor band at approximately 53 kD. Probing of overlapping synthetic peptides with pooled human cashew allergic sera identified 22 reactive peptides, 7 of which gave strong signals. Several Ana o 2 epitopes were shown to overlap those of the peanut legumin group allergen, Ara h 3, in position but with little sequence similarity. Greater positional overlap and identity was observed between Ana o 2 and soybean glycinin epitopes. CONCLUSIONS: We conclude that this legumin-like protein is a major allergen in cashew nut. PMID- 14555858 TI - Excretory-secretory products secreted by Paragonimus westermani delay the spontaneous cell death of human eosinophils through autocrine production of GM CSF. AB - BACKGROUND: Eosinophils play important roles in tissue inflammatory responses associated with helminth infections. Excretory-secretory products (ESP) produced by tissue-invasive helminths contain a large quantity of proteolytic enzymes that can modulate the host's immune responses. However, little is known regarding the roles of worm-derived products that are responsible for eosinophilic inflammatory responses in helminth infections. OBJECTIVE: In the present study, we investigated whether ESP produced by Paragonimus westermani, which cause pulmonary or extrapulmonary paragonimiasis in human beings, regulates both cell survival and death of human eosinophils. METHODS: The ESP was obtained from P. westermani newly excysted metacercariae (PwNEM). Eosinophils were purified from peripheral blood of healthy donors, and the purified eosinophils were incubated with or without the ESP secreted by PwNEM. The viability of eosinophils was assessed by staining with propidium iodide using the flow cytometer. RESULTS: When eosinophils were incubated with a low concentration of the ESP produced by PwNEM, which totally consists of proteolytic enzymes, eosinophil cell death was delayed compared with results for cells incubated with medium alone. In fact, the ESP at a low concentration stimulated eosinophils to produce detectable levels of GM-CSF that can delay eosinophil cell death. In contrast, eosinophil cell death was dose-dependently accelerated when cells were incubated with high concentrations of the ESP. To see whether the dose-dependent biphasic survival effect of the ESP on eosinophils is primarily due to the protease activity contained in the ESP, a high dose of the ESP was treated with heat at 56 degrees C for 30 min before being added to eosinophils. Attenuating protease activity in a high dose of the ESP by heat treatment reversed the ESP-afforded eosinophil cell death. This prolonged survival of eosinophils induced by the heated ESP was remarkably inhibited by anti-GM-CSF-neutralizing mAb and Jak2 kinase inhibitor AG 490. CONCLUSION: These results suggest that the proteases in the ESP secreted by PwNEM are able to regulate eosinophil survival through the autocrine production of GM-CSF. Thus, the enhanced eosinophil survival induced by Paragonimus-secreted products may contribute to the elicitation of eosinophilic inflammatory responses at the worm-infected lesion in human paragonimiasis. PMID- 14555859 TI - Antigenic specificity of serum antibodies in mice fed soy protein. AB - BACKGROUND: Soybean protein is used in a number of food products but unfortunately is also a common cause of food allergy. Upon ingestion of soy protein, healthy mice like other animals and humans generate a soy-specific antibody response in the absence of signs of illness. Not much is known about the relationship between the immunogenic proteins involved in this nondeleterious antibody response and the pathological response associated with food allergy. The objective of the present study was to characterize the antigenic specificity of the soy protein-specific antibody response generated in healthy mice ingesting soy protein. METHODS: Blood from mice fed a soy-containing diet was analyzed using ELISA and immunoblot for antibody reactivity towards various soy protein fractions and pure soy proteins/subunits. Mice bred on a soy-free diet were used as controls. RESULTS: The detectable antigenic specificity of the serum antibodies of soy-consuming mice comprised glycinin and beta-conglycinin. Immunoblots with soy protein extract demonstrated antibody reactivity towards both the basic and the acidic chains of glycinin and the beta-conglycinin subunits with an individual response pattern among mice. Moreover, antibody reactivity was found towards the native quaternary structure of glycinin. CONCLUSIONS: Mice ingesting soy protein generate an antibody response with reactivity towards glycinin and beta-conglycinin. Antibody reactivity found towards the native quaternary structure of glycinin indicates an oral immunogenicity of the highly processing-resistant oligomerized glycinin. PMID- 14555860 TI - Eosinophil count in nasal mucosa is more suitable than the number of ICAM-1 positive nasal epithelial cells to evaluate the severity of house dust mite sensitive allergic rhinitis: a clinical correlation study. AB - BACKGROUND: House dust mite (HDM)-sensitive allergic rhinitis is a perennial rhinitis with persistent nasal inflammation. Currently, there are no reliable parameters to monitor the severity of perennial allergic rhinitis. The purpose of this study was to evaluate correlations between clinical and laboratory parameters in patients with HDM-sensitive allergic rhinitis. METHODS: We measured nasal symptoms, did the Dermatophagoides pteronyssinus (Der P) skin prick test (SPT), evaluated the Der P allergen nasal challenge threshold, and laboratory parameters [(1) inflammatory cell count from nasal mucosal scraping specimens: eosinophils and neutrophils and (2) immunocytochemistry: ICAM-1 expression on nasal epithelial cells] in 20 cases of HDM-sensitive allergic rhinitis and performed correlation tests between all parameters. RESULTS: The wheal diameter induced by Der P SPT was significantly correlated with the Der P allergen nasal challenge threshold (p = 0.001). The number of eosinophils from nasal mucosal scrapping specimens was correlated with the ICAM-1 expression on nasal epithelial cells (p = 0.039), the number of neutrophils from nasal mucosal scrapping specimens (p = 0.001), and nasal stuffiness (p = 0.037) but did not correlate with total nasal symptom scores. CONCLUSION: Clinical symptoms of HDM-sensitive allergic rhinitis showed a poor correlation with inflammatory parameters. The eosinophil count in nasal mucosa is correlated with ICAM-1 expression and more suitable than ICAM-1 levels to evaluate the severity of HDM-sensitive allergic rhinitis. This study also supports the role of the SPT in the diagnosis of nasal allergy to HDM. PMID- 14555861 TI - Dissimilarity between seasonal changes in airway responsiveness to adenosine-5' monophosphate and methacholine in patients with grass pollen allergic rhinitis: relation to induced sputum. AB - BACKGROUND: In patients with allergic rhinitis, bronchial hyperresponsiveness (BHR) and airway inflammation may increase during pollen exposure. BHR can be assessed by adenosine-5'-monophosphate (AMP) or methacholine challenge. It has been suggested that BHR to AMP is more closely related to airway inflammation than BHR to methacholine. Seasonal allergic rhinitis offers a dynamic model to study changes in BHR and airway inflammation during natural allergen exposure. METHODS: We measured BHR [provocative concentration causing a 20% fall (PC(20)) in forced expiratory volume in 1 s (FEV(1))] to AMP and methacholine, and induced sputum cells in 16 rhinitis patients before and during the 2001 grass pollen season. RESULTS: There was a decrease in PC(20) methacholine during pollen exposure (geometric mean PC(20) from 3.22 to 1.73 mg/ml, p = 0.0023), whereas no reduction was observed for PC(20) AMP (p = 0.11). There was no increase in sputum eosinophils [pre: 0.69% (95% confidence interval 0.22-2.07); during: 1.85 (0.55- 5.6), p = 0.31]. Although the correlation of log PC(20) methacholine and log PC(20) AMP at baseline was good (r = 0.76, p = 0.001), individual seasonal changes (doubling concentrations) in PC(20) methacholine were not correlated with changes in PC(20) AMP (rho = 0.21, p = 0.44). There was no correlation between baseline log PC(20) methacholine or seasonal changes in PC(20) methacholine and sputum eosinophils (p > 0.12, all correlations). In contrast, there was a significant correlation between seasonal changes in PC(20) AMP and changes in sputum eosinophils (rho = -0.59, p = 0.025). CONCLUSIONS: These data show dissimilarity between seasonal changes in PC(20) AMP and methacholine in patients with seasonal allergic rhinitis. Moreover, PC(20) AMP seems to be more closely related to sputum eosinophils than PC(20) methacholine. The clinical significance of this discrepancy is unclear. PMID- 14555862 TI - Tolerability of rofecoxib in patients with adverse reactions to nonsteroidal anti inflammatory drugs: a study of 216 patients and literature review. AB - BACKGROUND: Adverse reactions to nonsteroidal anti-inflammatory drugs (NSAIDs) are frequent, particularly among patients with chronic urticaria or asthma. The identification of an alternative safe and reliable drug is a common problem in clinical practice. OBJECTIVE: To assess the tolerability of rofecoxib, a new NSAID that selectively inhibits the inducible isoform of cyclooxygenase, in a large group of NSAID-sensitive patients. METHODS: We studied 216 patients (164 females and 52 males) who had suffered adverse reactions to one or more NSAIDs; 98 subjects (45.4%) had experienced reactions to only one NSAID (single hypersensitivity) and 118 subjects (54.6%) had reacted to multiple NSAIDs (multiple hypersensitivity). Cutaneous reactions were reported by 79.6% of the subjects, respiratory symptoms by 10.7%, cutaneous and respiratory symptoms by 8.3%, anaphylaxis by 1.4%. All the subjects underwent a single-blind, placebo controlled oral challenge with divided therapeutic doses of rofecoxib (6.25 mg +18.75 mg 1 h later = cumulative dose of 25 mg). RESULTS: No reactions to the placebo were observed; only 1 subject (0.46%) experienced an urticarial reaction, after the second dose of rofecoxib. CONCLUSIONS: Considering previous studies and our own data, rofecoxib was well tolerated by all of the 174 patients with exclusively NSAID-related respiratory symptoms. Rofecoxib also had a very low rate (1.6%) of cross-reactivity in the 600 patients with exclusively cutaneous reactions to NSAIDs. PMID- 14555864 TI - Effects of legal mechanisms on perceived coercion and treatment adherence among persons with severe mental illness. AB - This study takes preliminary steps to examine the effects of 2 legal mechanisms outpatient commitment (OPC) and representative payeeship (rep payee)-on perceived coercion and treatment adherence in persons with severe mental illness (SMI). Data were collected in structured interviews from 258 involuntarily-admitted inpatients with SMI who were followed for 1 year after discharge. Subjects' clinicians and family members were also interviewed. The analyses show that subjects with both OPC and a rep payee perceived their mental health treatment to be significantly more coercive than subjects who did not receive these legal interventions. Duration of OPC and having a newly-appointed rep payee were both significantly associated with increased treatment adherence over the course of 1 year. For treatment-adherent subjects, neither form of legal mechanism was associated with perceived coercion in treatment, whereas subjects who were nonadherent perceived legal mechanisms as significantly more coercive. The data suggest that these legal tools both affect adherence to treatment, but when used together significantly increase the perceived coerciveness of treatment. Questions remain concerning the underlying mechanism of these effects. PMID- 14555863 TI - An association of maternal age and birth weight with end-stage renal disease in Saskatchewan. Sub-analysis of registered Indians and those with diabetes. AB - AIMS: To determine links between birth related factors and end-stage renal disease (ESRD). METHODS: This 1:3 age, sex, and source population (registered Indians [SkRI] and other Saskatchewan people [OSkP]) matched case-control study, compared maternal age and parity, gestational age, low birth weight (LBW), and high birth weight (HBW), between subjects with and without ESRD. RESULTS: Of 1,162 subjects, 277 cases (48 SkRI and 229 OSkP) and 601 controls (112 SkRI and 489 OSkP) had birth weight information. A trend for increased LBW rates occurred among SkRI and OSkP cases compared to controls (10.4 vs. 5.3% and 6.6 vs. 4.3%), and was significant for OSkP female cases (OR 3.66; 95% confidence interval [CI] 1.05, 12.73). Higher HBW rates occurred in SkRI cases (14.6% compared to 11.6% controls; N/S), and 3/5 female SkRI diabetic ESRD (DESRD) cases were over 3,750 g compared to 1/14 controls (p < 0.05). Only maternal age >/=30 years was an independent predictor for ESRD, particularly for OSkP non-DESRD cases (OR 2.45; 95% CI 1.03, 5.8). Cases with older mothers had lower mean birth weights than controls (3,236 vs. 3,434 g; p = 0.005). CONCLUSIONS: Older maternal age may predispose offspring to ESRD through mechanisms that differ for DESRD versus non DESRD, and that may relate to ethnicity. PMID- 14555865 TI - Sense of coherence: a mediator between violence exposure and psychopathology in Russian juvenile delinquents. AB - Previous research has demonstrated that exposure to community violence tends to have a hazardous impact on the health and well being of youth. In this context, sense of coherence is related to indicators of well being and mental health and is considered crucial for human information processing in resolving conflicts and coping with enduring stress. The aim of the present study was to explore the role of sense of coherence as a potential mediator in the relationships between community violence exposure and psychopathology, controlling for involvement in severe problem behavior. The study was conducted with a group of Russian juvenile delinquents (N = 159; a highly traumatized population with a history of frequent exposure to violence), who completed a set of questionnaires assessing their exposure to community violence, psychopathology, involvement in severe problem behaviors, and sense of coherence. The relationships among the variables of interest were explored using a structural equation modeling approach. Sense of coherence partially mediated the link between victimization and psychopathology. The direct relationships between victimization and psychopathology decreased in size, suggesting that higher levels of sense of coherence can potentially reduce the levels of psychopathology. These results have an important theoretical value and are meaningful for clinical work, suggesting that preventive and therapeutic interventions should aim at increasing sense of coherence, which might be especially valuable in populations at risk. PMID- 14555866 TI - The effect of early versus late antidepressant treatment on physical impairment associated with poststroke depression: is there a time-related therapeutic window? AB - Impairments in activities of daily living (ADL) are common after stroke and may be related to poststroke depression. We have demonstrated that remission of poststroke major depression was associated with improvement in ADL. The administration of antidepressants within the first 3 months after stroke has been shown to prevent poststroke depression, early administration might also improve recovery of ADL among patients with stroke. This study examines the effect of early versus late treatment with antidepressants on recovery in ADL. Among 62 patients after stroke, the therapeutic effect of a 3-month course of antidepressants begun during the first month after stroke was compared with the effect of treatment begun after 1 month. The severity of impairment was measured using the Functional Independence Measure (FIM) and post-treatment outcome was assessed over the following 21 months. Although both the early and late treatment groups showed improvements in FIM scores during the 3 months of treatment, the early treatment group improved significantly more than the late treatment group. After the treatment, the early treatment group maintained this improvement over 2 years while the late treatment group deteriorated over time. There were no significant differences in the 2 groups that would explain the findings. Recovery in ADL impairment after stroke appeared to be enhanced by the use of antidepressant medication if treatment was started within the first month after stroke. These findings are consistent with the hypothesis that there may be a time-related therapeutic window in the treatment of physical impairment associated with poststroke depression. PMID- 14555867 TI - Outcomes for depressed and anxious inpatients discharged before or after group cognitive behavior therapy: a naturalistic comparison. AB - Treatment outcomes for psychiatric inpatients with a primary diagnosis of a depressive or anxiety disorder who completed a cognitive behavior therapy (CBT) program while inpatients or when discharged were examined. Of 340 inpatients, 197 had been discharged to day patient status before the end of the CBT program, and 143 remained as inpatients. Data were collected before and after the CBT program and at 3-month follow-up. There was a significant improvement by posttreatment in self-esteem, locus of control, anxiety, depression, and stress, and these gains were maintained at 3-month follow-up. In addition, anxious, but not depressed, patients who completed CBT as day patients demonstrated greater improvements from after the CBT program to follow-up than those treated as inpatients. Possible reasons for the beneficial effects of completing the CBT as a day patient rather than an inpatient are discussed, and threats to the validity of this naturalistic study are considered. PMID- 14555868 TI - Subjective effects of antidepressants: a pilot study of the varieties of antidepressant-induced experiences in meditators. AB - The use of antidepressants continues to increase, yet relatively little is known about their precise subjective effects, and there is growing concern about subtle psychological side effects. One novel investigative approach to these problems may be to use introspectively trained subjects such as meditators. Experienced meditators recently taking antidepressants rated antidepressant effects on multiple dimensions of experience and reported significant emotional, motivational, and cognitive effects and benefits. This study suggests that a) meditators may benefit both clinically and meditatively from antidepressants, b) meditators may provide significant novel information on antidepressant effects, c) meditators may prove valuable for phenomenological investigations of psychopathology, drug effects, and therapies, d) meditation may prove a helpful maintenance therapy for depression, and e) enhanced equanimity may contribute to the broad therapeutic efficacy of antidepressants. PMID- 14555869 TI - Satisfaction with quality of life varies with temperament types of patients with schizophrenia. AB - We sought to explore the relationships of three temperament factors with domain specific subjective quality of life (QOL) of patients with schizophrenia. Ninety patients with schizophrenia were evaluated using the Quality of Life Enjoyment and Life Satisfaction Questionnaire, the Tridimensional Personality Questionnaire, the Positive and Negative Syndromes Scale, the Distress Scale for Adverse Symptoms, the Insight and Treatment Attitudes Questionnaire, the Insight Self-Report Scale, and standardized questionnaires for self-reported emotional distress and stress process-related variables. Predictors of domain-specific QOL were identified using multiple regression techniques. Temperament factors explain 6% to 16% of variability in QOL domain scores among patients with schizophrenia after controlling for the remaining variables (emotional distress, social support, self-esteem, avoidance coping, age, side effects, and depression). We found that higher levels of novelty seeking are associated with better general QOL, physical health, and more positive subjective feelings, whereas higher levels of reward dependence are related to better satisfaction from social relationships. Higher levels of harm avoidance are associated with poorer satisfaction with general activities, and medication. Thus, temperament factors, as assessed by the Tridimensional Personality Questionnaire, substantially influence satisfaction with life quality in schizophrenia. Novelty seeking, reward dependence, and harm avoidance are associated with different domains of QOL. PMID- 14555870 TI - Linking posttraumatic stress disorder and psychosis: a look at epidemiology, phenomenology, and treatment. AB - Several recent studies have provided direct evidence for the link between posttraumatic stress disorder (PTSD) and psychosis. Patients with psychotic disorders are known to be at a higher risk of traumatization and PTSD. Additionally, preclinical and clinical data suggest that the effects of trauma exposure on neural networks may provide a common diathesis for disorders like PTSD and schizophrenia. This article reviews evidence on a) the magnitude of association between PTSD and psychosis, b) the causal mechanisms implicated, and c) treatment considerations relevant to this association. A comprehensive MEDLINE search was conducted, and articles pertinent to epidemiological, clinical, and treatment aspects of comorbid PTSD and psychosis were identified. High rates of PTSD characterize patients with severe mental illness. Psychotic phenomena may also be a relatively common manifestation in patients with chronic PTSD. However, in clinical settings, the diagnosis is often missed, and few systematic guidelines exist for the identification and treatment of these comorbidities. Future neurobiological and treatment studies may be useful in better informing the clinical management of these subgroups. PMID- 14555871 TI - Posttraumatic stress disorder symptomatology in Vietnam veterans before and after September 11. PMID- 14555872 TI - Vigilance and avoidance during sleep in US Vietnam War veterans with posttraumatic stress disorder. PMID- 14555873 TI - Content of worry in the community: what do people with generalized anxiety disorder or other disorders worry about? PMID- 14555874 TI - The usefulness of the Axis V diagnosis: opinions of health care professionals. PMID- 14555877 TI - Tackling the growing global burden of atherosclerotic cardiovascular diseases. PMID- 14555878 TI - Type D personality, cardiac events, and impaired quality of life: a review. AB - BACKGROUND: Psychological distress has been associated with the pathogenesis and progression of coronary heart disease (CHD) but little is known about the determinants of distress as a coronary risk factor. Although it has become unfashionable to focus on personality factors since research on Type A behaviour yielded inconsistent findings, personality may comprise a major explanatory factor of individual differences in stress-related CHD. This article focuses on Type D--the distressed--personality, which describes patients who experience increased negative emotions and tend to inhibit the expression of these emotions in social interactions. METHODS: The article reviews research on Type D personality in the context of CHD. RESULTS: Accumulating evidence indicates that cardiac patients with the Type D personality are at increased risk for cardiovascular morbidity and mortality (odds ratios ranging from 4.1-8.9, P<0.0001) independent of standard cardiac risk factors. Type D patients are also at increased risk for psychological distress, clustering of psychosocial risk factors, impaired quality of life, and seem to benefit less from medical and invasive treatment. Preliminary evidence suggests that physiological hyper reactivity and activation of pro-inflammatory cytokines may be responsible for the detrimental effect of Type D personality on cardiac prognosis. CONCLUSIONS: There is an urgent need to adopt a personality approach in the identification of patients at risk for stress-related cardiac events. Type D is a stable personality construct that may be of special interest not only in CHD, but in other chronic cardiac conditions as well. PMID- 14555879 TI - Cardiovascular risk factor levels in urban and rural Thailand--The International Collaborative Study of Cardiovascular Disease in Asia (InterASIA). AB - BACKGROUND: Vascular mortality is increasing in economically developing countries but reliable data about the determinants of cardiovascular disease are few. The International Collaborative Study of Cardiovascular Disease in Asia (InterASIA) was designed to obtain precise estimates of cardiovascular risk factor levels in the adult population of Thailand. DESIGN: A complex sample survey. METHODS: Data from a structured questionnaire, brief physical examination and a blood sample were collected from 5305 individuals aged 35 years or older (response rate 68%). Mean risk factor levels were calculated for eight groups defined by age and sex in 18 representative urban and rural areas of Thailand. Population risk factor levels were calculated by applying sampling weights derived from the 2000 Thai Census and allowing for the complex sampling design. RESULTS: The estimated mean (standard error) population blood pressure was 120/76 (0.7/0.5) mmHg, mean serum total cholesterol was 5.2 (0.06) mmol/l, mean body mass index was 24 (0.2) kg/m2, mean fasting plasma glucose was 5.6 (0.06) mmol/l, the proportion with diabetes 9.6 (1)% and the proportion of current smokers was 25 (3)%. There were estimated to be 5.1 (0.5) million individuals with high blood pressure, 4.4 (0.4) million with high total cholesterol, 8.9 (0.8) million overweight or obese, 2.4 (0.2) million with diabetes and 6.2 (0.9) million current smokers. Mean levels of all major risk factors, except smoking, were worse in urban compared with rural areas. However, except for total cholesterol, the absolute numbers of individuals with abnormal risk factor levels were highest in rural areas. CONCLUSION: Absolute levels of cardiovascular risk factors in Thailand are high. Effective risk factor control strategies that target both rural and urban areas of Thailand have the potential to avert much premature cardiovascular disease. PMID- 14555880 TI - Psychosocial distress and impaired quality of life--targets neglected in the secondary prevention in women with ischaemic heart disease. AB - BACKGROUND: Secondary prevention in patients with ischaemic heart disease (IHD) focusing on classical risk factors has been less successful in women than in men. Previous studies have suggested that psychosocial factors may be new targets for secondary prevention. In the present study, psychosocial factors have been compared in women with and without IHD. METHODS AND SUBJECTS: Using a cross sectional comparison, classical cardiovascular risk factors and self-rated stress behaviour, vital exhaustion, stressful life events and quality of life were assessed in 198 women (age 35-77 years) who had IHD and a population-based sample of 206 women (age 45-74 years) without IHD. RESULTS: Control of classical risk factors was most often adequate in women with IHD. The prevalence of smoking was 9% in women with versus 18% in women without IHD (P<0.001), blood pressure levels were similar, and a very large share of women with IHD were on treatment with anti-thrombotic, anti-hypertensive and/or lipid-lowering drugs (P<0.001 compared with women free of IHD). In contrast, women with IHD scored significantly worse than women without IHD in self-rated stress behaviour (P=0.003 after adjustment for other possible determinants), and in vital exhaustion (P=0.003). In univariate analyses, quality of life was significantly worse in women with IHD. Stressful life events did not differ between the two groups. CONCLUSIONS: In this group of women with IHD, control of classical risk factors was good, whereas control of psychosocial risk factors was clearly inadequate. Secondary prevention by psychosocial intervention in women with IHD should be evaluated in randomized controlled trials. PMID- 14555881 TI - A critical appraisal of the prognostic value of the VE/VCO2 slope in chronic heart failure. AB - BACKGROUND: Increased ventilatory drive, reflected by the slope of increase of ventilation relative to carbon dioxide production (VE/VCO2 slope), has been shown to have a high prognostic value in patients with chronic heart failure (CHF). However, there is no consensus on how to calculate it, as the relation between VE and VCO2 becomes nonlinear near the end of exercise, when ventilation is driven both by CO2 output and by decrease in plasma pH. This may influence the results. METHODS: Ninety-seven CHF patients with ejection fraction <45% (mean 27 +/- 9%), in NYHA class II-IV underwent a cardiopulmonary exercise test. VE/VCO2 slope was assessed by linear regression using all the data points (Sl), using only points before (Sli), or after (Slf) the non-linear part of the curve, and using only the first 3-min data (Sl3 min). Peak oxygen uptake (VO2), and circulatory power (VO2 x systolic arterial pressure) were also assessed. Death and transplantation were the end-points considered (mean follow-up 22 months). RESULTS: Mean value of VE/VCO2 overall slope was 39.3 +/- 11.6 (22-78). In 64% of the patients, two distinct slopes could be found: an initial, linear slope (31.8 +/- 7.5, 18-62) and a final, steeper slope (48.6 +/- 15.7, 24-101). Patients in whom no rupture of slope was observed were sicker. There was a relation between initial and overall VE/VCO2 slopes (r=0.915, P<10(-4)) and between overall and 3-min VE/VCO2 slopes (r=0.808, P<10(-4)). VE/VCO2 slope correlated with peak VO2 (r=-0.55, P<10(-4)) and peak circulatory power (r=-0.49, P<10(-4)). Univariate analysis showed that the prognostic value of overall VE/VCO2 slope (chi2 25.4, P<10(-4)) was greater than initial (chi2 22.8, P<10(-4)), 3-min (chi2 14.6, P<10(-4)) or final VE/VCO2 slopes (chi2 6.7, P=0.09). By multivariate analysis, the prognostic value of the peak circulatory power was similar to that of the VE/VCO2 overall slope. CONCLUSIONS: The VE/VCO2 slope should be computed from all the data points to have its highest prognostic value. Peak circulatory power also has similar prognostic value. PMID- 14555882 TI - Nuclear factor-kappa B activation in skeletal muscle of patients with chronic heart failure: correlation with the expression of inducible nitric oxide synthase. AB - BACKGROUND: In the advanced stages of chronic heart failure (CHF) the expression of inducible nitric oxide synthase (iNOS) in skeletal muscle (SM) may contribute to exercise intolerance and early fatigue. Cell culture studies and promoter analysis demonstrated that the transcription factor nuclear factor kappa B (NF kappaB) is essential for iNOS-expression. The aim of this study was to assess whether NF-kappaB is activated in skeletal muscle of patients with CHF and linked to the expression of iNOS. METHODS: Skeletal muscle biopsies were obtained from seven CHF patients and seven healthy controls (HC). Nuclear proteins were isolated and the content of activated NF-kappaB was analysed by electrophoretic mobility shift assay (EMSA). Inducible nitric oxide synthase expression in SM was determined by real time polymerase chain reaction (PCR). RESULTS: The expression of iNOS (1.4 +/- 0.2 versus 0.5 +/- 0.08 arbitrary units, P=0.001) and the activation of NF-kappaB in the SM (0.5 +/- 0.1 versus 0.1 +/- 0.04 arbitrary units, P=0.009) was significantly increased in CHF patients as compared to healthy controls. Furthermore, a linear correlation was observed between NF kappaB activation and iNOS expression (r=0.78, P<0.001). CONCLUSIONS: The results of this study indicate for the first time that in the skeletal muscle of patients with chronic heart failure the activation of the transcription factor NF-kappaB is increased and may represent one important regulatory factor for the expression of iNOS in patients. Nevertheless, due to the small sample size this observation has to be confirmed in subsequent studies with more patients. PMID- 14555883 TI - A prospective study of infection and cardiovascular diseases: the Busselton Health Study. AB - BACKGROUND: Infectious agents might play a role in the aetiology of cardiovascular diseases. The aim was to determine the association of antibodies to implicated infectious agents with coronary heart disease (CHD) and stroke in a population-based prospective study. DESIGN: This study was based on a cohort of 1612 cardiovascular disease-free adults in the 1981 Busselton Health Survey. Primary risk factors were measured from stored serum and case-cohort sampling was used to reduce costs and preserve serum. The outcomes of interest were time to first CHD or stroke event. Serum antibody tests were carried out for all 218 CHD cases, all 119 stroke cases and a random subset of 451 subjects. METHODS: Sera were tested for antibodies to Chlamydia pneumoniae (IgG and IgA), and for IgG antibodies to Helicobacter pylori and cytomegalovirus (CMV). The association between serum antibody and risk of cardiovascular diseases was analysed using Cox proportional hazards regression. RESULTS: The estimated population prevalence was 24% for C. pneumoniae IgG, 7% for C. pneumoniae IgA, 58% for H. pylori and 85% had CMV antibody levels greater than 15 AU/mL. The estimated relative risk of CHD was around 1.2 for all antibodies examined, except for C. pneumoniae IgA for which it was less than one, and the estimated relative risk of stroke was around 0.85, however in all cases the 95% confidence interval included one. CONCLUSIONS: This study of an Australian population does not support an association between serum antibody levels to C. pneumoniae, H. pylori and CMV with development of cardiovascular diseases. PMID- 14555884 TI - Insulin resistance and other risk factors for coronary heart disease in elderly men. The Study of Men Born in 1913 and 1923. AB - BACKGROUND: We have previously shown that insulin is not a major risk factor for CHD. Our data have been re-analysed with longer follow-up and the homeostasis model assessment (HOMA) method. DESIGN: Prospective cohort study of 57- and 67 year-old men. METHODS: Insulin resistance was estimated with the HOMA equation. Standard methods were used to measure risk factors. The endpoint (CHD) was the combination of non-fatal myocardial infarction and fatal CHD during 13 years of follow-up. RESULTS: The risk of CHD increased 2.5-fold in known diabetics compared with those with normal glucose tolerance, 2.2-fold among those in the highest compared with the lowest quintile of insulin resistance, and 2.4-fold among those in the highest compared with the lowest quintile of fasting insulin. Increased physical activity decreased the risk of CHD by 65%. Cholesterol was also a significant risk factor for CHD but blood pressure, obesity and smoking were not related to the incidence of CHD. CONCLUSION: Insulin resistance is a significant risk factor for CHD. Whether it is a causal risk factor remains to be proven. Regular physical activity protects against CHD. PMID- 14555885 TI - Predictive value of classical risk factors and their control in coronary patients: a follow-up of the EUROASPIRE I cohort. AB - BACKGROUND: Both EUROASPIRE studies revealed the suboptimal management of coronary patients regarding lifestyle changes and prophylactic use of cardiovascular drugs. We report here on the mortality follow-up of the EUROASPIRE I cohort over a median period of 4.4 years. DESIGN AND METHODS: The EUROASPIRE I cohort consisted of a consecutive sample of patients aged < or =70 years from nine European countries, hospitalized because of coronary artery bypass graft, percutaneous transluminal coronary angioplasty, acute myocardial infarction or myocardial ischaemia. Baseline data, gathered in 1995-96 through standardized methods, were linked to cause-specific mortality as registered up to 1 April 2000 in 3343 patients. RESULTS: After adjustment for age, gender and diagnostic category according to Cox modelling, smoking, previous coronary heart disease and diabetes proved significant predictors of total, cardiovascular (CVD) and coronary heart disease (CHD) mortality. Obesity, low education, raised blood pressure, elevated total cholesterol and low HDL cholesterol, however, were not significantly associated with higher mortality rates. In multivariate analysis, smoking and diabetes emerged as the strongest predictors of CVD [risk ratios (RR) 2.2 and 2.5 respectively] and CHD mortality (RR 2.4 and 2.4 respectively). CONCLUSIONS: The results of the mortality follow-up of the EUROASPIRE I patients underline the importance of smoking and diabetes in the secondary prevention of CHD. Failure to find statistically significant associations between other classical risk factors, such as blood pressure and plasma lipid levels, and mortality may be related to the extensive use of antihypertensive and lipid lowering drugs in this cohort. PMID- 14555887 TI - Exercise prescription based on heart rate: a simple thing or a science? PMID- 14555886 TI - Influence of beta-blocker use on percentage of target heart rate exercise prescription. AB - BACKGROUND: Exercise is recommended for cardiac patients irrespective of beta blockers. Percentages of maximal heart rate (%HRmax) and heart rate reserve (%HRR) are widely used to determine training intensities. The purpose of this study was to investigate the influence of chronic cardioselective beta blockade on the %HRmax and %HRR model. METHODS: Ten healthy male subjects randomly received oral placebo or beta-blocker bisoprolol (5 mg/day) for 2 weeks using a double-blind, crossover design. In the second week, the subjects performed a cardiopulmonary exercise test until exhaustion to determine the aerobic (AeT) and anaerobic (AnT) threshold. RESULTS: No significant differences were found for absolute and relative values of oxygen consumption, power output and ratings of perceived exertion at AeT, AnT and maximum workload. Mean HR was significantly (P<0.05) lower at rest (-15 +/- 5 bpm), AeT (-19 +/- 8 bpm), AnT (-22 +/- 10 bpm) and maximal workload (-19 +/- 11 bpm) with bisoprolol compared to placebo. Percentage of maximal heart rate (%HRmax) was significantly (P<0.05) reduced at rest (43 versus 39%), AeT (64 versus 60%) and AnT (86 versus 82%), a trend for a reduction was found for %HRR at AnT (75 versus 71%, P=0.07). CONCLUSIONS: Exercise prescription using %HRmax or %HRR methods are of limited accuracy for patients taking beta-blockers. Although %HRmax and %HRR are easy to determine and therefore attractive, we suggest that the most precise exercise prescription would depend on AeT and AnT. Percentages of maximal oxygen consumption or maximal workload or ratings of perceived exertion may be suggested as a substitute. Alternatively, upper limits for %HRmax and %HRR should be lower for patients taking beta-blockers. PMID- 14555888 TI - How to carry out a health-orientated marathon training programme for running and inline skating. AB - BACKGROUND: More and more people want to do something for their health. Nevertheless, certain reluctance exists when it comes to committing oneself to higher sporting aims, as well as some uncertainty concerning proper training. DESIGN AND METHODS: Two model projects have been developed, aimed at motivating larger parts of the population towards a health-orientated training programme. Both projects serve as a basis for further, similarly designed large-scale projects. In the running project a health-focused run training programme was set up aimed at preparing people, previously inexperienced in running, for a marathon. In the inline project an inline training programme was developed which comprised both the training of essential techniques as well as individualized endurance training, the aim being the participation in an inline marathon. RESULTS: A total of more than 1000 people participated in both projects. The health check performed before the beginning of the project revealed some pathological findings, demonstrating the necessity of a medical examination before taking up regular training. Both projects led to an improvement in endurance performance. In addition, the blood-chemical metabolic parameters showed clear optimization, in particular the lipid profile. The inline technique training resulted in an improvement of the braking skills, which is accompanied by a reduction in the risk of injuries. CONCLUSIONS: Both model projects have shown that large-scale sports programmes that focus primarily on the prevention of injuries and disease (under sport-medical guidance) with a performance incentive are accepted by the population, provided they offer a high fun factor. PMID- 14555889 TI - European guidelines on cardiovascular disease prevention in clinical practice: third joint task force of European and other societies on cardiovascular disease prevention in clinical practice (constituted by representatives of eight societies and by invited experts). AB - Guidelines aim to present all the relevant evidence on a particular issue in order to help physicians to weigh the benefits and risks of a particular diagnostic or therapeutic procedure. They should be helpful in everyday clinical decision-making. A great number of guidelines have been issued in recent years by different organisations--European Society of Cardiology (ESC), American Heart Association (AHA), American College of Cardiology (ACC), and other related societies. By means of links to web sites of National Societies several hundred guidelines are available. This profusion can put at stake the authority and validity of guidelines, which can only be guaranteed if they have been developed by an unquestionable decision-making process. This is one of the reasons why the ESC and others have issued recommendations for formulating and issuing guidelines. In spite of the fact that standards for issuing good quality guidelines are well defined, recent surveys of guidelines published in peer reviewed journals between 1985 and 1998 have shown that methodological standards were not complied with in the vast majority of cases. It is therefore of great importance that guidelines and recommendations are presented in formats that are easily interpreted. Subsequently, their implementation programmes must also be well conducted. Attempts have been made to determine whether guidelines improve the quality of clinical practice and the utilisation of health resources. In addition, the legal implications of medical guidelines have been discussed and examined, resulting in position documents, which have been published by a specific task force. The ESC Committee for practice guidelines (CPG) supervises and coordinates the preparation of new guidelines and expert consensus documents produced by task forces, expert groups or consensus panels. The Committee is also responsible for the endorsement of these guidelines or statements. PMID- 14555890 TI - Over the counter cosmetic colored contact lenses: deja vu (disaster!) all over again! PMID- 14555891 TI - Ocular complications associated with the use of cosmetic contact lenses from unlicensed vendors. AB - PURPOSE: To call attention to the unauthorized sale of cosmetic contact lenses, resulting in ocular complications. DESIGN Observational case report. METHODS: Retrospective, observational, clinical practice setting. RESULTS: Six patients (five female and one male) were seen urgently for acute eye pain and redness after wearing cosmetic plano contact lenses. None of the patients had previously worn a contact lens or spectacle correction. None of the lenses were dispensed by eye care professionals. One patient developed pseudomonal keratitis, ultimately requiring penetrating keratoplasty for visual rehabilitation. CONCLUSIONS: Colored contact lenses are being dispensed without a prescription or fitting from unlicensed vendors, such as cosmetics, convenience, and accessory stores. Lenses are sold individually and without care instructions. Consequently, uninformed lens wearers are experiencing acute, vision-threatening infections and inflammation. PMID- 14555892 TI - Kersley lecture: eye believe in contact lenses: contact lenses and/or refractive surgery. AB - PURPOSE: Media publicity has made patients increasingly aware that surgical vision correction is available. This article gives an overview on refractive surgery and contact lenses. METHODS: Contact lens and refractive surgery possibilities are highlighted for patients with myopia, hyperopia, astigmatism, presbyopia, aphakia, and keratoconus. Therapeutic, pediatric, and cosmetic indications are discussed. RESULTS: Refractive surgery is beneficial for low refractive errors. The use of contact lenses is mandatory in cases of monovision and strabismus before refractive surgery and beneficial for therapeutic use postoperatively. CONCLUSION: Although many patients are always best corrected with contact lenses, only the right synergism of contact lenses and refractive surgery will benefit patients. PMID- 14555893 TI - Ulcerative keratitis in contact lens wearers. AB - PURPOSE: To determine the clinical microbiological characteristics of corneal ulcers in contact lens wearers. METHODS: A retrospective study of 23 patients admitted to our department with contact lens-related corneal ulcers during a 43 month period. Detailed demographic data, the type of contact lens, duration of lens wear, and wearing schedule were derived from a self-administered questionnaire. The severity of the ulcer; cultures of corneal scrapings, storage solutions, and contact lenses; treatment; and final outcome were evaluated. RESULTS: Of the 86 cases of ulcerative keratitis admitted during the study period, 23 (26.74%) were attributed to contact lens use. Most patients were young women from urban areas. All of them were using soft contact lenses for 3 days to 20 years. Five patients used daily-wear lenses as extended-wear lenses. Most ulcers (47.82%) were mild; 30.43% were moderate; and 21.47% were severe. Corneal scrapings for cultures were obtained in 15 of the cases and were positive in 10 (43.47%) of them, whereas in 33.33% of the culture-positive storage solutions and in 66.67% of the culture-positive contact lenses, corneal scrapings were negative. Pseudomonas aeruginosa was the most frequent isolated pathogen (60%). The final visual acuity was 20/40 or better in 60.87% of the cases. CONCLUSIONS: Contact lens use is an important risk factor for the development of ulcerative keratitis, with P. aeruginosa remaining the predominant pathogen. It seems important to culture contact lenses and contact lens storage solutions, in addition to the corneal scrapings, and the role of initial therapy for the corneal ulcers remains important. PMID- 14555894 TI - Do the economic and social factors play an important role in relation to the compliance of contact lenses care routines? AB - PURPOSE: To evaluate the behavior of contact lens wearers of distinct economic and social groups in relation to their contact lens care routines. METHODS: The authors studied 60 patients of distinct social groups at the same Contact Lens Department at Sorocaba Eye Hospital in Sorocaba, Brazil. The patients were divided in two groups. Group 1 included 31 patients who could not pay for medical assistance and were in the Government Health Assistance Program. Group 2 included 29 patients who had health insurance and patients paying for medical assistance. We studied their age, sex, period of contact lens wear, and type of contact lenses that they were wearing and related these variables to the contact lens care routines. RESULTS: There was no significant difference between the two groups in relation to their contact lens care routines. CONCLUSIONS: Social factors do not seem to be the most important factor in the compliance of contact lens care routines. PMID- 14555895 TI - Evaluation of corneal staining and patient preference with use of three multi purpose solutions and two brands of soft contact lenses. AB - PURPOSE: Clinical signs and symptoms vary depending on contact lens brand, lens care products, or a combination of the two. Studies evaluated corneal staining and subject preference associated with three multipurpose solutions (MPS) and two soft contact lens brands. METHODS: Two sites conducted the randomized, investigator-masked, 2-month crossover studies. Forty-five subjects used regimen 1 (No Rub Opti-Free Express MPS [Alcon, Fort Worth, TX]) and regimen 2 (Complete MPS [Allergan, Irvine, CA]) for 1 month each (study 1). In study 2, 44 subjects used regimen 1 and regimen 3 (ReNu MultiPlus MPS [Bausch & Lomb, Rochester, NY]). ACUVUE 2 (Johnson & Johnson Vision Care, Jacksonville, FL) or SofLens 66 (Bausch & Lomb) lenses were randomly assigned to subjects for the duration of the study. Corneal staining was evaluated on days 0 (baseline), 7, 14, and 28 of each month. RESULTS: Corneal staining was significantly worse with regimen 3 for both lens brands at day 28 compared with baseline (P < 0.01). Significant differences in staining were noted between regimens 1 and 3 throughout the study, primarily with SofLens 66 lenses (P or =20% of primary tumours) at CASP8, SLIT2 and RASSF1A in Wilms' tumour and at RASSF1A, TIMP3, DAPK, SLIT2, MT1G and GSTP1 in RCC. PMID- 14555993 TI - Murine pancreatic tumor cell line TD2 bears the characteristic pattern of genetic changes with two independently amplified gene loci. AB - TGFalpha/p53(+/-) transgenic mice represent a genetically engineered mouse model for pancreatic adenocarcinoma. The tumors develop a characteristic pattern of secondary genetic changes. From one of these tumors, the permanent cell line TD2 was established. Here, we describe in detail the genetic changes by molecular cytogenetic techniques. The original tumor-specific CGH profile has been retained unchanged. The most characteristic aberration pattern bears chromosome 11. Egfr, localized on proximal chromosome 11, is amplified two to three times and leads to an easily identifiable, stable marker chromosome with a large amplification unit, which is present in each metaphase. The wild-type p53 gene on distal chromosome 11 is lost. The p16Ink4a locus on chromosome 4 is hypermethylated. For c-Myc a 15 fold amplification, present in a 1.65 Mb amplification unit, is detected on chromosome 15. Transition between presence in the form of several double minutes, DMs, or a single homogeneously staining region, HSR, was observed for c-Myc. Molecular-cytogenetic analysis of both amplification units show that Egfr amplification and c-Myc amplification represent two alternative modes by which genes get amplified in tumor cells. The expression level of the respective genes was proven by Northern blot analysis. The cell line TD2 represents a valuable in vitro model for pancreatic adenocarcinoma. PMID- 14555994 TI - Molecular subclassification of kidney tumors and the discovery of new diagnostic markers. AB - We analysed the expression profiles of 70 kidney tumors of different histological subtypes to determine if these subgroups can be distinguished by their gene expression profiles, and to gain insights into the molecular mechanisms underlying each subtype. In all, 39 clear cell renal cell carcinomas (RCC), seven primary and one metastatic papillary RCC, six granular RCC from old classification, five chromophobe RCC, five sarcomatoid RCC, two oncocytomas, three transitional cell carcinomas (TCC) of the renal pelvis and five Wilms' tumors were compared with noncancerous kidney tissues using microarrays containing 19,968 cDNAs. Based on global gene clustering of 3560 selected cDNAs, we found distinct molecular signatures in clear cell, papillary, chromophobe RCC/oncocytoma, TCC and Wilms' subtypes. The close clustering in each of these subtypes points to different tumorigenic pathways as reflected by their histological characteristics. In the clear cell RCC clustering, two subgroups emerged that correlated with clinical outcomes, confirming the potential use of gene expression signatures as a predictor of survival. In the so-called granular cell RCC (terminology for a subtype that is no longer preferred), none of the six cases clusters together, supporting the current view that they do not represent a single entity. Blinded histological re-evaluation of four cases of 'granular RCC' led to their reassignment to other existing histological subtypes, each compatible with our molecular classification. Finally, we found gene sets specific to each subtype. In order to establish the use of some of these genes as novel subtype markers, we selected four genes and performed immunohistochemical analysis on 40 cases of primary kidney tumors. The results were consistent with the gene expression microarray data: glutathione S-transferase alpha was highly expressed in clear cell RCC, alpha methylacyl racemase in papillary RCC, carbonic anhydrase II in chromophobe RCC and K19 in TCC. In conclusion, we demonstrated that molecular profiles of kidney cancers closely correlated with their histological subtypes. We have also identified in these subtypes differentially expressed genes that could have important diagnostic and therapeutic implications. PMID- 14555995 TI - X-ray crystal structure of IRF-3 and its functional implications. AB - Transcription factor IRF-3 is post-translationally activated by Toll-like receptor (TLR) signaling and has critical roles in the regulation of innate immunity. Here we present the X-ray crystal structure of the C-terminal regulatory domain of IRF-3(175-427) (IRF-3 175C) at a resolution of 2.3 A. IRF-3 175C is structurally similar to the Mad homology domain 2 of the Smad family. Structural and functional analyses reveal phosphorylation-induced IRF-3 dimerization, which generates an extensive acidic pocket responsible for binding with p300/CBP. Although TLR and Smad signaling are evolutionarily independent, our results suggest that IRF-3 originates from Smad and acquires its function downstream of TLR. PMID- 14555996 TI - Crystal structure of IRF-3 reveals mechanism of autoinhibition and virus-induced phosphoactivation. AB - IRF-3, a member of the interferon regulatory factor (IRF) family of transcription factors, functions as a molecular switch for antiviral activity. IRF-3 uses an autoinhibitory mechanism to suppress its transactivation potential in uninfected cells, and virus infection induces phosphorylation and activation of IRF-3 to initiate the antiviral responses. The crystal structure of the IRF-3 transactivation domain reveals a unique autoinhibitory mechanism, whereby the IRF association domain and the flanking autoinhibitory elements condense to form a hydrophobic core. The structure suggests that phosphorylation reorganizes the autoinhibitory elements, leading to unmasking of a hydrophobic active site and realignment of the DNA binding domain for transcriptional activation. IRF-3 exhibits marked structural and surface electrostatic potential similarity to the MH2 domain of the Smad protein family and the FHA domain, suggesting a common molecular mechanism of action among this superfamily of signaling mediators. PMID- 14555997 TI - Tandem PDZ repeats in glutamate receptor-interacting proteins have a novel mode of PDZ domain-mediated target binding. AB - The interaction of the glutamate receptor subunits 2 and 3 (GluR2/3) with multi PDZ domain glutamate receptor-interacting protein (GRIP) is important for the synaptic trafficking and clustering of the receptors. Binding of GluR2/3 to GRIP requires both the fourth and fifth PDZ domains (PDZ4 and PDZ5) to be covalently linked, although only one PDZ domain is directly involved in binding to the receptor tail. To elucidate the molecular basis of this mode of PDZ domain mediated target recognition, we solved the solution structures of the PDZ45 tandem and the isolated PDZ4 of GRIP. The two PDZ domains form a compact structure with a fixed interdomain orientation. The interdomain packing and the stable folding of both PDZ domains require a short stretch of amino acids N terminal to PDZ4 and a conserved linker connecting PDZ4 and PDZ5. PDZ4 contains a deformed aB-bB groove that is unlikely to bind to carboxyl peptides. Instead, the domain stabilizes the structure of PDZ5. PMID- 14555998 TI - Structural correspondence between the alpha-helix and the random-flight chain resolves how unfolded proteins can have native-like properties. AB - Recently, we have proposed that, on average, the structure of the unfolded state of small, mostly alpha-helical proteins may be similar to the native structure (the 'mean-structure' hypothesis). After examining thousands of simulations of both the folded and the unfolded states of five polypeptides in atomistic detail at room temperature, we report here a result that seems at odds with the mean structure hypothesis. Specifically, the average inter-residue distances in the collapsed unfolded structures agree well with the statistics of the ideal random flight chain with link length of 3.8 A (the length of one amino acid). A possible resolution of this apparent contradiction is offered by the observation that the inter-residue distances in a typical alpha-helix over short stretches are close to the average distances in an ideal random-flight chain. PMID- 14556000 TI - Liquid-liquid phase transition in supercooled silicon. AB - Silicon in its liquid and amorphous forms occupies a unique position among amorphous materials. Obviously important in its own right, the amorphous form is structurally close to the group of 4-4, 3-5 and 2-6 amorphous semiconductors that have been found to have interesting pressure-induced semiconductor-to-metal phase transitions. On the other hand, its liquid form has much in common, thermodynamically, with water and other 'tetrahedral network' liquids that show density maxima. Proper study of the 'liquid-amorphous transition', documented for non-crystalline silicon by both experimental and computer simulation studies, may therefore also shed light on phase behaviour in these related materials. Here, we provide detailed and unambiguous simulation evidence that the transition in supercooled liquid silicon, in the Stillinger-Weber potential, is thermodynamically of first order and indeed occurs between two liquid states, as originally predicted by Aptekar. In addition we present evidence to support the relevance of spinodal divergences near such a transition, and the prediction that the transition marks a change in the liquid dynamic character from that of a fragile liquid to that of a strong liquid. PMID- 14556001 TI - Interfacial heat flow in carbon nanotube suspensions. AB - The enormous amount of basic research into carbon nanotubes has sparked interest in the potential applications of these novel materials. One promising use of carbon nanotubes is as fillers in a composite material to improve mechanical behaviour, electrical transport and thermal transport. For composite materials with high thermal conductivity, the thermal conductance across the nanotube matrix interface is of particular interest. Here we use picosecond transient absorption to measure the interface thermal conductance (G) of carbon nanotubes suspended in surfactant micelles in water. Classical molecular dynamics simulations of heat transfer from a carbon nanotube to a model hydrocarbon liquid are in agreement with experiment. Our findings indicate that heat transport in a nanotube composite material will be limited by the exceptionally small interface thermal conductance (G approximately 12 MW m(-2) K(-1)) and that the thermal conductivity of the composite will be much lower than the value estimated from the intrinsic thermal conductivity of the nanotubes and their volume fraction. PMID- 14556002 TI - Hepatocyte growth factor and its receptor are required for malaria infection. AB - Plasmodium, the causative agent of malaria, must first infect hepatocytes to initiate a mammalian infection. Sporozoites migrate through several hepatocytes, by breaching their plasma membranes, before infection is finally established in one of them. Here we show that wounding of hepatocytes by sporozoite migration induces the secretion of hepatocyte growth factor (HGF), which renders hepatocytes susceptible to infection. Infection depends on activation of the HGF receptor, MET, by secreted HGF. The malaria parasite exploits MET not as a primary binding site, but as a mediator of signals that make the host cell susceptible to infection. HGF/MET signaling induces rearrangements of the host cell actin cytoskeleton that are required for the early development of the parasites within hepatocytes. Our findings identify HGF and MET as potential targets for new approaches to malaria prevention. PMID- 14556003 TI - Essential role of endothelial nitric oxide synthase for mobilization of stem and progenitor cells. AB - Endothelial nitric oxide synthase (eNOS) is essential for neovascularization. Here we show that the impaired neovascularization in mice lacking eNOS is related to a defect in progenitor cell mobilization. Mice deficient in eNOS (Nos3(-/-)) show reduced vascular endothelial growth factor (VEGF)-induced mobilization of endothelial progenitor cells (EPCs) and increased mortality after myelosuppression. Intravenous infusion of wild-type progenitor cells, but not bone marrow transplantation, rescued the defective neovascularization of Nos3(-/ ) mice in a model of hind-limb ischemia, suggesting that progenitor mobilization from the bone marrow is impaired in Nos3(-/-) mice. Mechanistically, matrix metalloproteinase-9 (MMP-9), which is required for stem cell mobilization, was reduced in the bone marrow of Nos3(-/-) mice. These findings indicate that eNOS expressed by bone marrow stromal cells influences recruitment of stem and progenitor cells. This may contribute to impaired regeneration processes in ischemic heart disease patients, who are characterized by a reduced systemic NO bioactivity. PMID- 14556004 TI - TRAM is specifically involved in the Toll-like receptor 4-mediated MyD88 independent signaling pathway. AB - Recognition of pathogens by Toll-like receptors (TLRs) triggers innate immune responses through signaling pathways mediated by Toll-interleukin 1 receptor (TIR) domain-containing adaptors such as MyD88, TIRAP and TRIF. MyD88 is a common adaptor that is essential for proinflammatory cytokine production, whereas TRIF mediates the MyD88-independent pathway from TLR3 and TLR4. Here we have identified a fourth TIR domain-containing adaptor, TRIF-related adaptor molecule (TRAM), and analyzed its physiological function by gene targeting. TRAM-deficient mice showed defects in cytokine production in response to the TLR4 ligand, but not to other TLR ligands. TLR4- but not TLR3-mediated MyD88-independent interferon-beta production and activation of signaling cascades were abolished in TRAM-deficient cells. Thus, TRAM provides specificity for the MyD88-independent component of TLR4 signaling. PMID- 14556005 TI - Tim-3 inhibits T helper type 1-mediated auto- and alloimmune responses and promotes immunological tolerance. AB - Although T helper (T(H)) cell-mediated immunity is required to effectively eliminate pathogens, unrestrained T(H) activity also contributes to tissue injury in many inflammatory and autoimmune diseases. We report here that the T(H) type 1 (T(H)1)-specific Tim-3 (T cell immunoglobulin domain, mucin domain) protein functions to inhibit aggressive T(H)1-mediated auto- and alloimmune responses. Tim-3 pathway blockade accelerated diabetes in nonobese diabetic mice and prevented acquisition of transplantation tolerance induced by costimulation blockade. These effects were mediated, at least in part, by dampening of the antigen-specific immunosuppressive function of CD4(+)CD25(+) regulatory T cell populations. Our data indicate that the Tim-3 pathway provides an important mechanism to down-regulate T(H)1-dependent immune responses and to facilitate the development of immunological tolerance. PMID- 14556006 TI - Interaction of Tim-3 and Tim-3 ligand regulates T helper type 1 responses and induction of peripheral tolerance. AB - T helper type 1 (T(H)1) immune responses are central in cell-mediated immunity, and a T(H)1-specific cell surface molecule called Tim-3 (T cell immunoglobulin domain, mucin domain) has been identified. Here we report the identification of a secreted form of Tim-3 that contains only the immunoglobulin (Ig) variable (V) domain of the full-length molecule. Fusion proteins (Tim-3-Ig) of both Tim-3 isoforms specifically bound CD4(+) T cells, indicating that a Tim-3 ligand is expressed on CD4(+) T cells. Administration of Tim-3-Ig to immunized mice caused hyperproliferation of T(H)1 cells and T(H)1 cytokine release. Tim-3-Ig also abrogated tolerance induction in T(H)1 cells, and Tim-3-deficient mice were refractory to the induction of high-dose tolerance. These data indicate that interaction of Tim-3 with Tim-3 ligand may serve to inhibit effector T(H)1 cells during a normal immune response and may be crucial for the induction of peripheral tolerance. PMID- 14556007 TI - Tat-binding protein-1, a component of the 26S proteasome, contributes to the E3 ubiquitin ligase function of the von Hippel-Lindau protein. AB - von Hippel-Lindau (VHL) gene inactivation occurs in von Hippel-Lindau (VHL) disease. The protein pVHL functions in a multi-subunit E3 ubiquitin ligase that targets the hypoxia-inducible transcription factor Hif1 alpha for proteasomal degradation during normoxia. We establish that pVHL binds to Tat-binding protein 1 (TBP-1), a component of the 19S regulatory complex of the proteasome. TBP-1 associates with the beta-domain of pVHL and complexes with pVHL and Hif1 alpha in vivo. Overexpression of TBP-1 promotes degradation of Hif1 alpha in a pVHL dependent manner that requires the ATPase domain of TBP-1. Blockade of TBP-1 expression by small interfering RNA (siRNA) causes prolonged degradation kinetics of Hif1 alpha. Several distinct mutations in exon 2 of VHL disrupt binding of pVHL to TBP-1. A pVHL mutant containing a P154L substitution coimmunoprecipitates with Hif1 alpha, but not TBP-1, and does not promote degradation of Hif1 alpha. Thus, the ability of pVHL to degrade Hif1 alpha depends in part on its interaction with TBP-1 and suggests a new mechanism for Hif1 alpha stabilization in some pVHL-deficient tumors. PMID- 14556008 TI - Mutations in a novel gene encoding a CRAL-TRIO domain cause human Cayman ataxia and ataxia/dystonia in the jittery mouse. AB - Cayman ataxia is a recessive congenital ataxia restricted to one area of Grand Cayman Island. Comparative mapping suggested that the locus on 19p13.3 associated with Cayman ataxia might be homologous to the locus on mouse chromosome 10 associated with the recessive ataxic mouse mutant jittery. Screening genes in the region of overlap identified mutations in a novel predicted gene in three mouse jittery alleles, including the first mouse mutation caused by an Alu-related (B1 element) insertion. We found two mutations exclusively in all individuals with Cayman ataxia. The gene ATCAY or Atcay encodes a neuron-restricted protein called caytaxin. Caytaxin contains a CRAL-TRIO motif common to proteins that bind small lipophilic molecules. Mutations in another protein containing a CRAL-TRIO domain, alpha-tocopherol transfer protein (TTPA), cause a vitamin E-responsive ataxia. Three-dimensional protein structural modeling predicts that the caytaxin ligand is more polar than vitamin E. Identification of the caytaxin ligand may help develop a therapy for Cayman ataxia. PMID- 14556009 TI - Plasma endothelin-1 in patients with stable or unstable angina. AB - BACKGROUND: It has been documented that an elevated endothelin-1 (ET-1) plasma concentration is associated with an increased risk of serious coronary events and the presence of angiographically documented coronary artery disease (CAD). The results of a few studies which examined ET-1 plasma level in patients with stable or unstable angina, were inconclusive. AIM: To assess whether ET-1 blood concentration measured in the coronary sinus and peripheral vein is associated with clinical symptoms in patients with multi-vessel CAD. METHODS: The study group consisted of 23 patients with multi-vessel CAD of whom 11 had unstable angina and 12 - stable angina. Both groups were matched with regard to age, gender and the presence of cardio-vascular risk factors. Blood samples for ET-1 assessment were taken during coronary angiography simultaneously from the coronary sinus and femoral vein. ET-1 was measured using an immunoenzymatic method. RESULTS: ET-1 plasma level in the peripheral venous circulation was similar in patients with unstable or stable angina (0.45+/-0.18 pmol/L versus 0.46+/-0.14 pmol/L, NS) whereas ET-1 level in the coronary sinus was significantly higher in patients with unstable angina (1.44+/-0.47 pmol/L versus 0.34+/-0.17 pmol/L, p<0.05). CONCLUSIONS: ET-1 concentration in the coronary sinus is significantly higher in patients with unstable rather than stable angina which confirms the role of ET-1 in the pathogenesis of CAD. Our results suggest a possible future role of endothelin receptor blockers in the treatment of patients with unstable angina. PMID- 14556010 TI - Myocardial infarction in patients with diabetes. Results of primary coronary angioplasty. AB - BACKGROUND: Although the introduction of primary percutaneous coronary interventions (PCI) improved the outcome of patients with acute myocardial infarction (MI), diabetes remains a significant factor which worsens prognosis. AIM: To compare the immediate and in-hospital results of PCI in patients with acute MI with or without diabetes. METHODS: The outcome of 139 patients with diabetes and 528 patients without diabetes was compared. Thrombolytic therapy was administered prior to PCI to 43.2% of patients with diabetes and 42.4% of patients without diabetes. RESULTS: Patients with diabetes were older, more frequently of female gender and had higher incidence of hypertension as well as multi-vessel coronary artery disease. PCI was effective in 85.6% of diabetics and 90.2% of non-diabetic patients (NS). The reocclusion rate was significantly higher in diabetics than in non-diabetics (11.5% vs 5.5%, p=0.012) whereas the incidence of haemorrhagic complications was similar. Mortality rates were comparable in both groups (7.2% in diabetics vs 5.9% in non-diabetics, NS). CONCLUSIONS: 1) Immediate efficacy of primary PCI for acute MI is similar in diabetics and non-diabetics, however, the in-hospital reocclusion rate is higher in the former group of patients. 2) In-hospital mortality is not affected by the presence of diabetes. 3) Thrombolytic and invasive treatment of diabetic patients with acute MI is not associated with an increased risk of bleeding. PMID- 14556011 TI - Beta-thromboglobulin and platelets in unstable angina. AB - BACKGROUND: Atheromatous plaque rupture is the main cause of platelet activation in ischaemic heart disease (IHD). Platelet activation is manifested by a release into circulation of the components of granules, including beta-thromboglobulin (beta-TG) - a marker of platelet activation in vivo. The platelet count (PLT), mean platelet volume (MPV) and the proportion of large platelets (L(PLT)) are indirect platelet activation markers. Data in literature on the role of these markers in patients with unstable angina are discordant. AIM: To assess plasma concentration of beta-TG, PLT, MPV and LPLT in patients with unstable angina before and during standard pharmacological therapy. METHODS: The study group consisted of 54 patients (19 females and 35 males) with unstable angina who were divided into two groups: Group A - 45 patients with a history of angina, and group B - nine patients with a new onset unstable angina. beta-TG and platelet activation markers were measured at baseline (groups A and B) and after 8-10 days of standard medical therapy for unstable angina (group B). The control group consisted of 26 healthy subjects (13 females and 13 males). RESULTS: The mean beta-TG concentration in groups A (16.2 IU/ml) and B (19.7 IU/ml - before and 21.8 IU/ml - after treatment) was significantly (p<0.05) higher than in controls (10.6 IU/ml). In patients with unstable angina, the PLT and MPV values were not affected by therapy and were similar to those obtained in controls, whereas the LPLT value was significantly higher than in controls. CONCLUSIONS: Concentrations of beta-TG and L(PLT) are increased in patients with unstable angina due to platelet activation. The introduction of standard medical treatment for unstable angina did not significantly change beta-TG and platelet activation markers. PMID- 14556012 TI - Is preinfarction angina associated with better outcome after myocardial infarction? AB - BACKGROUND: In a clinical setting of acute myocardial infarction (MI), short lasting and transient anginal pain, preceding the development of acute MI, is regarded as a symptom representing ischaemic preconditioning. Some experimental and clinical data suggested that preinfarction angina may favourably influence the course of acute MI. AIM: We sought to examine the hypothesis that preinfarction angina occurring within 24 hours prior to the onset of acute MI favourably influences the outcome. METHODS: The study group consisted of 331 patients who were admitted to our hospital due to acute MI with ST segment elevation with a symptom duration <12 hours and received thrombolysis. Preinfarction angina within 24 hours prior to MI was present in 80 patients whereas the remaining 251 patients had no chest pain preceding acute MI. The course of the in-hospital phase of MI (mean 15 days) was analysed. RESULTS: In patients with preinfarction angina the in-hospital complication rate was significantly lower than in patients without angina preceding acute MI (p<0.001). Patients without preinfarction angina more frequently developed heart failure (p<0.001) or died (p<0.01) in hospital. Patients with preinfarction angina had significantly less extensive MI and had reperfusion symptoms more frequently. Multivariate analysis showed that there were three factors which independently favourably influenced survival: preinfarction angina (p=0.01), age < or =65 years (p=0.04) and duration of chest pain during acute MI < or =3h (p=0.03). Of the analysed group, 73 patients died in hospital. The independent variables predicting death included prior MI (p=0.04), history of diabetes (p=0.02), acute left bundle branch block (p=0.01) and age >65 years (p=0.03). Non-fatal re infarction complicated the in-hospital course of MI in 27 patients. The independent variables which predicted this complication included age >65 years (p=0.03) and hypercholesterolemia (p=0.04). CONCLUSIONS: Patients with preinfarction angina, occurring within 24 hours of acute MI, have better in hospital outcome and less extensive myocardial injury than patients without antecedent angina. These results may be attributed to the protective effects of ischaemic preconditioning. PMID- 14556014 TI - [Acute, massive pulmonary embolism treated with surgical embolectomy without cardio-pulmonary by-pass - a case report]. AB - A case of a 44-year-old male with massive pulmonary embolism is presented. Due to recent intra-cranial haemorrhage and on-going urinary bleeding, the patient was not treated with anticoagulant or thrombolytic agents. Because of the symptoms of cardiogenic shock, an urgent surgical pulmonary embolectomy was performed, using an approach firstly described by Trendelenburg in 1908. The procedure was performed without a cardio-pulmonary by-pass which was contra-indicted in this patient. During surgery, a massive thrombus from both right and left pulmonary arteries was removed. The patient survived surgery and seven days later was transferred to another hospital where the rehabilitation process was successfully continued. PMID- 14556013 TI - Percutaneous internal carotid artery angioplasty with stenting: early and long term results. AB - BACKGROUND: Cerebro-vascular accidents are the third most common cause of death. The most frequent localisation of lesions responsible for stroke are bifurcation of the common carotid artery or the proximal internal carotid artery (ICA). Surgical carotid endarterectomy (CEA) and carotid artery stenting (CAS) are the non-pharmacological methods used to treat carotid artery stenosis. AIM: To assess the efficacy and safety of CAS of ICA. METHODS: CAS was performed in 75 patients (49 males, 26 females) with a mean age of 65.2+/-9.1 years. Twenty (26.7%) patients underwent CAS with the use of the central nervous system (CNS) protective devices. The immediate, mid-term, and long-term results were analysed. RESULTS: In total, 84 stents were implanted to 80 ICA in 73 patients. In two patients stent implantation was not possible. In 7 (9%) patients with a stenosis of both ICA, a bilateral procedure was performed. In two patients concomitant dilatations of the vertebral artery, and in the other two - of subclavian artery, were performed. In 38 patients coronary angiography was performed directly before CAS; one patient underwent coronary angioplasty. In 20 patients protective CNS devices were used. During the procedure four patients developed ischaemic stroke on the side of CAS. In one patient neurological symptoms completely disappeared within 48 hours. The type of technique used during CAS did not influence the frequency of ischaemic complications. Four patients developed hyperperfusion syndrome which disappeared after a few days. There were fire deaths during follow up: three due to myocardial infarction (MI), one - after urgent CABG, and one due to pulmonary embolism. There were no deaths due to stroke. No new ischaemic changes in CNS nor significant changes in the neurological status, using the UNSS or Barthel scales, were observed. Asymptomatic restenosis was documented in six patients whereas one patient developed symptomatic restenosis due to stent deformation. CONCLUSIONS: Percutaneous angioplasty of an internal carotid artery carries a risk not exceeding that of surgical endarterectomy. In our study, a one year follow-up revealed a minor risk of ischaemic stroke. Percutaneous angioplasty with the use of protective devices should be tested in larger groups of patients in order to establish the real clinical usefulness and improved safety of this technique. PMID- 14556015 TI - [Primary coronary angioplasty with stent implantation in a patient with dextrocardia - a case report]. AB - A case of a 45-year-old male with acute myocardial infarction and dextrocardia is presented. The patient underwent successful primary coronary angioplasty with direct stenting. Difficulties in establishing diagnosis and treating patients with dextrocardia and chest pain are discussed. PMID- 14556016 TI - [Aortic valve stenosis when to operate?]. PMID- 14556020 TI - [Electrocardiogram of the month]. PMID- 14556021 TI - [Fistula between aorta and left atrium due to bacterial endocarditis in a patient with biological aortic valve - a case report]. PMID- 14556022 TI - [Angiogram of the month]. PMID- 14556023 TI - [Ventricular bigeminy originating from right ventricular outflow tract - treated with RF ablation]. PMID- 14556024 TI - [Cardiovascular rehabilitation in the era of invasive cardiology]. PMID- 14556026 TI - A trend of improved survival of childhood hepatoblastoma treated with cisplatin and doxorubicin in Taiwanese children. AB - Hepatoblastoma is the second most common childhood malignant hepatic tumor in Taiwan. Its prognosis used to be poor. We reviewed our cases in this decade to see if there has been any improvement of survival in our patients with hepatoblastoma. From 1988 to 2000, 19 patients with hepatoblastoma in this institution were included in the study. These patients' clinical manifestations, laboratory and image studies, histological findings, treatment modalities and prognostic significance were analyzed. The mean age at diagnosis was 13.5 months, ranging from 0 to 4 years old (male:female =11:8). Abdominal distension was the most common symptom, and hepatomegaly was the most common physical finding. Laboratory abnormalities included elevated alpha-fetoprotein, thrombocytosis and abnormal liver function profiles. Treatment modalities included primary surgery with postoperative chemotherapy in three, chemotherapy only in four and preoperative chemotherapy plus surgery with or without postoperative chemotherapy in nine patients. The overall 2-year survival rate is 38.6%. The significant prognostic factors include patients' compliance, resectability and chemotherapy protocol. The introduction of a new chemotherapy protocol designed by the International Society of Pediatric Oncology Study (SIOPEL) in 1994 improved the 2 year survival rate from 12.5 to 58.4% ( P=0.01). In conclusion, the improved chemotherapy protocol enhances the survival rate of hepatoblastoma in Taiwanese children. PMID- 14556027 TI - Enteral glutamine supplementation and dexamethasone attenuate the local intestinal damage in rats with experimental necrotizing enterocolitis. AB - The pathogenesis of necrotizing enterocolitis (NEC) presumptively is due to an inappropriate intestinal epithelial immunologic response of immature gut to luminal stimuli. Glutamine is essential for intestinal crypt cell proliferation and enhances the cellular response to growth factors. We aimed to test the hypothesis that the supplementation of enteral feedings with glutamine may stimulate an immature intestine and decrease the intestinal inflammatory change in NEC. Immediately after birth, the neonatal rats were weighed and randomized into one of four treatment groups. Group 1 consisted of rats whom were breast fed. Group 2 (NEC group) consisted of neonates whom were fed with a special rodent formula. Rats in groups 3 and 4 were fed in a similar fashion to those in group 2, and glutamine 0.3 mg/kg per day and dexamethasone 0.5 mg/kg per day were added to their formula, respectively. The neonatal rats were weighed and killed on day 4: the last 4 cm of terminal ileum was harvested for morphological studies and detection of nitrite and nitrate levels in tissue. The animals in the NEC group showed various degrees of inflammatory changes similar to clinical NEC. The inflammatory changes of the intestine appeared to be attenuated in both glutamine and steroid-treated animals compared to those in the NEC group. Only steroid treatment decreased the tissue levels of these nitrogen oxides that were increased in rats in the NEC group. We herein provide evidence that maturational agents such as glutamine and dexametasone can attenuate the local intestinal inflammatory damage in experimental NEC. These findings support the hypothesis that the gut immaturity in premature infants represents a risk factor for NEC. PMID- 14556028 TI - A new indication for buccal mucosal graft: isolated congenital fistula of the penile urethra. AB - Isolated congenital urethrocutaneous fistula is uncommon, and its repair has been associated with high incidence of recurrence. However, the use of buccal mucosal graft offers a satisfactory closure after previous failures. We report a new case in whom we adopted the buccal mucosal urethral replacement to treat the recurrence. PMID- 14556029 TI - Familial posterior fossa arachnoid cyst. AB - BACKGROUND: Arachnoid cysts are a relatively common incidental finding on CT scans of the brain. They most commonly occur in the middle cranial fossa, where familial occurrence has rarely been reported. Posterior fossa arachnoid cysts are more unusual. CASE HISTORIES: We report the presence of quadrigeminal cistern arachnoid cysts in siblings. PMID- 14556030 TI - Progressive posthemorrhagic hydrocephalus leads to changes of amplitude integrated EEG activity in preterm infants. AB - AIM: Intraventricular hemorrhage (IVH) is the most common cause of brain lesions in preterm infants. Among infants with IVH about 35% develop posthemorrhagic hydrocephalus (PPH) which may lead to secondary injury. Therapeutic interventions to reduce the increased intracranial pressure are invasive and carry a high risk of complications. Amplitude-integrated EEG (aEEG) allows continuous neurophysiological surveillance and may help in defining the optimal timing for intervention in infants with progressive PHH. In this report we show, for the first time, a change in aEEG activity in two preterm infants with PHH. METHODS: Cerebral activity was continuously monitored by aEEG provided by the Cerebral Function Monitor (Lectromed, UK) in two preterm infants with PPH. RESULTS: With increasing ventricular width, aEEG showed an increased discontinuity without distinguishable sleep-wake cycling in both infants. One infant showed an abrupt onset of a nearly isoelectric pattern without any change in clinical condition. Clinical signs of increased intracranial pressure developed 6-12 h later in both children. In one patient, aEEG activity returned to normal after successful shunting and reduction of intracranial pressure. CONCLUSION: Continuous neurophysiological monitoring by aEEG may be of value in the diagnostic and therapeutic management of preterm infants with progressive PHH. PMID- 14556031 TI - Source localization of the re-build up phenomenon in pediatric moyamoya disease-a dipole distribution analysis using MEG and SPECT. AB - OBJECTS: The "re-build up" phenomenon is a pathognomonic finding induced after hyperventilation in pediatric moyamoya disease, and is closely related to ischemic attacks. However, its precise mechanism remains unknown. METHODS: In this study, MEG, MRI, and SPECT were used to clarify its mechanism in 4 children with moyamoya disease. Cerebral blood flow before and after acetazolamide administration was measured. Spontaneous magnetic brain activity was measured using a whole-head MEG system. The quantified dipole distribution was shown on 3D MRI. RESULTS AND CONCLUSION: The original current dipoles of the "re-build up" slow waves were predominantly located in the deep cortical sulci in the area with impaired reactivity to acetazolamide. The results suggested that the re-build up waves derive from the deep cortical sulci due to the reduction in the cerebral perfusion reserve. PMID- 14556033 TI - Low-contrast detectability in volume rendering: a phantom study on multidetector row spiral CT data. AB - To cope with the increasing amount of CT data, there is growing interest in direct volume-rendering techniques (VRT) as a diagnostic tool. The aim of this phantom study was to analyze the low-contrast detectability (LCD) of VRT compared with multi-planar reformations (MPR). Soft tissue lesions were simulated by spheres of different diameters (3-8 mm). The average lesion density was 15 HU compared with a background density of 35 HU. Two different CT protocols with 40 and 150 mAs were performed on a multi-detector row CT. The scanning parameters were as following: 140 kV; 2x0.5-mm slice collimation; pitch 2 (table movement per rotation/single slice collimation), and reconstruction with 0.5-mm slice thickness at 0.5-mm interval. A B30 kernel was used for reconstruction. The VRT was performed by mapping Hounsfield values to gray levels equal to a CT window (center: 60 HU; window: 370 HU ). A linear ramp was applied for the opacity transfer function varying the maximum opacity between 0.1 and 1.0. A statistical method based on the Rose model was used to calculate the detection threshold depending on lesion size and image noise. Additionally, clinical data of 2 patients with three liver lesions of different sizes and density were evaluated. In VRT, LCD was most dependent on object size. Regarding lesions larger than 5 mm, VRT is significantly superior to MPR (p<0.05) for all opacity settings. In lesions sized 3-5 mm a maximum opacity level approximately 40-50% showed a near equivalent detectability in VRT and MPR. For higher opacity levels VRT was superior to MPR. Only for 3-mm lesions MPR performed slightly better in low contrast detectability (p<0.05). Compared with MPR, VRT shows similar performance in LCD. Due to noise suppression effects, it is suited for visualization of data with high noise content. PMID- 14556032 TI - Perspectives and limitations of image-guided neurosurgery in pediatric patients. AB - INTRODUCTION: Progress in image-guided neurosurgery, and specifically in computer assisted frameless navigation techniques and the application of robotic systems, has brought about many changes in the way we approach and treat pathologies involving the adult and pediatric central nervous system (CNS). Nevertheless, children are a patient group with special demands, in whom image-guided surgical techniques have certain limitations. In this article we address the common problem of registration accuracy in image-guided neurosurgery and assess both the reliability and the potential of ultrasound-based neuronavigation and robot assisted neuroendoscopy in pediatric patients. METHODS AND RESULTS: We present our experience with new referencing and tracking techniques adapted to the needs of the pediatric patient. In addition, advantages and limitations of a 3D real time ultasonography-based navigation system are demonstrated. Finally, we describe our experience with robot-assisted neuroendocopy and discuss the possible implications of the technique for the future. CONCLUSION: Image-guided techniques in pediatric neurosurgery are valuable tools and may open up new perspectives in the future. From our point of view, these tools enhance both intellectual and manual surgical skills, which does not mean that they should be considered as a replacement for such skills. PMID- 14556034 TI - Hyperpolarized 3-helium MR imaging of the lungs: testing the concept of a central production facility. AB - The aim of this study was to test the feasibility of a central production facility with distribution network for implementation of hyperpolarized 3-helium MRI. The 3-helium was hyperpolarized to 50-65% using a large-scale production facility based at a university in Germany. Using a specially designed transport box, containing a permanent low-field shielded magnet and dedicated iron-free glass cells, the hyperpolarized 3-helium gas was transported via airfreight to a university in the UK. At this location, the gas was used to perform in vivo MR experiments in normal volunteers and patients with chronic obstructive lung diseases. Following initial tests, the transport (road-air-road cargo) was successfully arranged on six occasions (approximately once per month). The duration of transport to imaging averaged 18 h (range 16-20 h), which was due mainly to organizational issues such as working times and flight connections. During the course of the project, polarization at imaging increased from 20% to more than 30%. A total of 4 healthy volunteers and 8 patients with chronic obstructive pulmonary disease were imaged. The feasibility of a central production facility for hyperpolarized 3-helium was demonstrated. This should enable a wider distribution of gas for this novel technology without the need for local start-up costs. PMID- 14556035 TI - Bone mineral density in patients with Behcet's disease. AB - Behcet's disease is a complex, multisystemic, inflammatory disorder characterized clinically by recurrent oral and genital ulcerations as well as uveitis, sometimes leading to blindness. The etiology and pathogenesis of this syndrome remain obscure. However, various factors are suspected, including genetic propensity, infectious precipitants, and immunological abnormalities. Considering the chronicity and unclear etiology of the disease, we conducted a prospective investigation of a possible alteration in the bone mineral density of affected persons. Thirty-five patients (18 males and 17 females, mean age 38.02+/-7.93 years) diagnosed with Behcet's disease and 33 sex- and age-matched healthy controls (14 males and 19 females, mean age 40.06+/-7.66 years) were seen on an outpatient basis, and bone densitometry measurements were done from June 2000 to December 2002 at the Mersin University Hospital in Turkey. Postmenopausal women with Behcet's disease and patients receiving oral corticosteroid therapy were excluded from the study. The mean disease duration was 6.68+/-7.05 years. Bone mineral density was measured with dual X-ray absorptiometry at the lumbar spine and right femur. The mean Z scores of the patient and control groups were -0.50+/ 1.06 and -0.13+/-0.92 at the lumbar spine, respectively, and 0.38+/-1.07 and 0.45+/-1.20 at the right femur, respectively. No significant differences in bone mineral density values were detected in the groups at either the lumbar (P = 0.15) or right femur (P = 0.82) site. Body mass index and disease duration did not influence bone mineral density, and age had a positive correlation with bone mineral density in patients with Behcet's disease. In conclusion, although it is difficult to draw definite conclusions due to the relatively small sample size, our study confirms that bone mineral density in Behcet's disease was not lower than in healthy subjects. PMID- 14556036 TI - A delta-endotoxin encoded in Pseudomonas fluorescens displays a high degree of insecticidal activity. AB - The short field-life of Bacillus thuringiensis (Bt) insecticidal crystal protein has limited its use. When the Bt toxin is produced in Pseudomonas fluorescens it can be encapsulated and retain its effectiveness for two to three times longer than other Bt formulations. In order to improve Bt expression, we have synthesized cryIA(c) Bt delta-endotoxin encoding region (GenBank AF537267) according to the usage codon of P. fluorescens and transformed the Bt toxin expression cassette into P. fluorescens strains. T7 RNA polymerase and the T7 promoter system were used to control expression of Bt toxin. SDS-PAGE and Western blotting assay revealed that the delta-endotoxin was expressed as 8% of the total protein in P. fluorescens. In in vitro tests, release of toxin from dead bacteria was demonstrated. Supplementation of diets with Bt toxin-containing Pseudomonas bacterium resulted in high mortality of cabbage butterfly ( Pieris brassicae) larvae. PMID- 14556037 TI - Effect of carbon monoxide, hydrogen and sulfate on thermophilic (55 degrees C) hydrogenogenic carbon monoxide conversion in two anaerobic bioreactor sludges. AB - The conversion routes of carbon monoxide (CO) at 55 degrees C by full-scale grown anaerobic sludges treating paper mill and distillery wastewater were elucidated. Inhibition experiments with 2-bromoethanesulfonate (BES) and vancomycin showed that CO conversion was performed by a hydrogenogenic population and that its products, i.e. hydrogen and CO2, were subsequently used by methanogens, homo acetogens or sulfate reducers depending on the sludge source and inhibitors supplied. Direct methanogenic CO conversion occurred only at low CO concentrations [partial pressure of CO (PCO) <0.5 bar (1 bar=10(5) Pa)] with the paper mill sludge. The presence of hydrogen decreased the CO conversion rates, but did not prevent the depletion of CO to undetectable levels (<400 ppm). Both sludges showed interesting potential for hydrogen production from CO, especially since after 30 min exposure to 95 degrees C, the production of CH4 at 55 degrees C was negligible. The paper mill sludge was capable of sulfate reduction with hydrogen, tolerating and using high CO concentrations (PCO>1.6 bar), indicating that CO-rich synthesis gas can be used efficiently as an electron donor for biological sulfate reduction. PMID- 14556038 TI - Metabolic analysis of acetate accumulation during xylose consumption by Paenibacillus polymyxa. AB - Paenibacillus polymyxa ATCC 12321 produced more acetic acid and less butanediol from xylose than from glucose. The product yields from xylose were ethanol (0.72 mol/mol sugar), (R,R)-2,3-butanediol (0.31 mol/mol sugar), and acetate (0.38 mol/mol sugar) while those from glucose were ethanol (0.74 mol/mol sugar), (R,R) 2,3-butanediol (0.46 mol/mol sugar), and acetate (0.05 mol/mol sugar). Higher acetate kinase activity and lower acetate uptake ability were found in xylose grown cells than in glucose-grown cells. Furthermore, phosphoketolase activity was higher in xylose-grown cells than in glucose-grown cells. In fed-batch culture on xylose, glucose feeding raised the butanediol yield to 0.56 mol/mol sugar and reduced acetate accumulation to 0.04 mol/mol sugar. PMID- 14556039 TI - Homogeneous batch cultures of Aspergillus oryzae by elimination of wall growth in the Variomixing bioreactor. AB - A novel principle for mixing and aeration in stirred bioreactors, named Variomixing, was developed. Four baffles are rotated intermittently at a rotational speed slower or similar to the speed of a centrally placed axial flow impeller. Rotational speeds of the baffles and impeller of 5-10 and 500-600 rpm, respectively, results in the highly turbulent flow regime characteristic of conventional bioreactors with high mixing and mass transfer capacities. Stagnant zones around crevices and crannies in which wall growth may commence are avoided since the baffles are never completely at rest. Increasing the rotational speed of the baffles (5 s every 5 min), so that it follows the speed of the impeller (500-600 rpm), cancels the effect of the baffles and a deep vortex and high peripheral liquid flow rates at the reactor wall develop. The vortex ensures that also the head-space of the reactor wall is flushed and any deposits removed. The filamentous fungus Aspergillus oryzae has been grown in batch cultures in the Variomixing bioreactor. Compared to conventional laboratory-scale bioreactors, in which more than 30% of all biomass was found attached to walls, less than 2% of the total A. oryzae biomass was found on the walls in the Variomixing bioreactor. PMID- 14556040 TI - Characterization of the ask-asd operon in aminoethoxyvinylglycine-producing Streptomyces sp. NRRL 5331. AB - The first two genes of the threonine pathway, ask and asd, were cloned and sequenced from the aminoethoxyvinylglycine-producing Streptomyces sp. NRRL 5331. The two genes are organized in a bicistronic operon. ask, encoding the apartokinase (ASK), is located upstream from asd. The presence of a ribosome binding site within the ask sequence suggests that this open reading frame encodes two overlapping proteins. The formation of both subunits of the aspartokinase from a single gene was studied using antibodies raised against the C-terminal end of the aspartokinase subunits. Disruption of asd results in a significant decrease of aminoethoxyvinylglycine production, thus supporting the involvement of the ask-asd operon in the biosynthesis of this metabolite. This is the first report in which a gene cluster for the first two steps of aminoethoxyvinylglycine biosynthesis is characterized. PMID- 14556041 TI - Collagenolytic proteases from bacteria. AB - Collagen degradation occurs during various physiological and pathological conditions. However, only a limited number of proteases with unique characteristics can trigger collagen degradation. Until recently, practical applications of collagenolytic proteases from bacteria had not been considered because their functions in bacteria are closely related to pathogenicity. However, bacterial collagenolytic proteases have many interesting and useful features. This review focuses on the collagenolytic proteases from bacteria, in particular their molecular properties and practical applications. PMID- 14556042 TI - Perception of the risk of gastrointestinal adverse drug reactions with non steroidal anti-inflammatory drugs (including coxibs): differences among general practitioners, gastroenterologists and rheumatologists. AB - OBJECTIVE: To compare the perception of risk of gastrointestinal adverse drug reactions (ADRs) with non-steroidal anti-inflammatory drugs (NSAIDs) (including coxibs) in general practitioners, gastroenterologists and rheumatologists. METHODS: Physicians (69 general practitioners, 45 gastroenterologists and 58 rheumatologists; 172 total) were interviewed using visual analogue scales in order to define a score of perceived risk (ranking from 0 to 10) of gastrointestinal ADRs associated with four classes of NSAIDs (indolics, oxicams, arylcarboxylic derivatives, coxibs). RESULTS: When the three groups of physicians were taken as a whole, indolic derivatives were ranked as the most dangerous NSAIDs [median score: 5.3 (25-75th percentiles: 3.5-7)] followed by oxicams [median score: 5.0 (25-75th percentiles: 3.1-6.2)], arylcarboxylic derivatives [median score 4.7 (25-75th percentiles: 3.0-6.5)] and coxibs [median score 1.9 (25-75th percentiles: 1.2-3.0)]. Among physicians, general practitioners gave the most important mean score [median score 4.8 (25-75th percentiles: 3.4-5.7)] for NSAIDs as a whole followed by gastroenterologists [median score 4.6 (25-75th percentiles: 3.6-5.6)] and rheumatologists [median score 3.6 (25-75th percentiles: 2.3-4.7)]. There was no significant difference among the three groups of physicians in the median score of perceived digestive risk associated with the use of indolic derivatives, whereas significant differences were found for arylcarboxylic derivatives, oxicams and coxibs, with higher median scores given by gastroenterologists followed by general practitioners and rheumatologists. CONCLUSION: This study shows differences in the perception of risks of gastrointestinal ADRs. Rheumatologists systematically considered NSAIDs as less harmful than general practitioners and gastroenterologists did. In contrast, the widely demonstrated difference in gastrointestinal risk among the different groups of NSAIDS (with, for example, a higher risk for oxicams) was not pointed out by the three studied groups of physicians. There are differences in the perception of gastrointestinal risks of NSAIDs (including coxibs) among physicians according to their medical education and medical specialisation. PMID- 14556043 TI - Preconcentration of selenium compounds on a porous graphitic carbon column in view of HPLC-ICP-AES speciation analysis. AB - The retention of organic selenium compounds on a porous graphitic carbon stationary phase was investigated. Different acids were studied as mobile phases to elute selenocystamine, selenoethionine, selenomethionine and selenocystine. Detection was achieved using inductively coupled plasma-atomic emission spectrometry to provide selenium-specific and sensitive detection. The separation of the four species was carried out using methanoic acid. An important on-column preconcentration was obtained when solutes were injected in nitric acid or trifluoroacetic acid (TFA) media. The large injection volume employed (2,500 micro L) allowed us to reach low relative detection limits (2-6 micro g/L). The method, employing TFA as injection solvent and methanoic acid as the eluent was found to be robust with respect to different matrices spiked with selenocompounds. PMID- 14556044 TI - Rapid determination of telmisartan in pharmaceuticals and serum by the parallel catalytic hydrogen wave method. AB - The polarographic characteristics of telmisartan have been investigated in 0.8 mol L(-1) NH3.H2O-NH4Cl (pH 8.9)-0.01 mol L(-1) H2O2 as supporting electrolyte. The results demonstrate that the polarographic reduction wave at ca. -1.30 V in the absence of H2O2 is a catalytic hydrogen wave, and the reduction wave enhanced by H2O2 is a so-called parallel catalytic hydrogen wave. The analytical sensitivity of the parallel catalytic hydrogen wave is ca. 60 times higher than that of the corresponding catalytic hydrogen wave. Based on the parallel catalytic hydrogen wave a novel method has been developed for determination of telmisartan by linear sweep polarography. The calibration curve is linear in the range 2.0x10(-8)-2.0x10(-6) mol L(-1) and the detection limit is 1.0x10(-8) mol L(-1). The precision is excellent with relative standard deviations of 2.6% at a concentration of 1.0x10(-7) mol L(-1) telmisartan. The proposed method has been applied to the direct determination of the telmisartan in capsule forms and biological samples. The proposed method has been proved to be advantageous over existing CZE and MEKC methods in simplicity, rapidity, and reproducibility. PMID- 14556045 TI - Analysis of silica hydride and surface hydrosilation reactions by solid-state NMR in the preparation of p-chlorobenzamide bonded silica phase. AB - 29Si and (13)C CP-MAS NMR spectroscopy was used to follow the conversion of native silica to a p-chlorobenzamide bonded silica material. The benzamide bonded phase was prepared via a hydrosilation reaction of a hydride silica intermediate with p-chloro- N-allylbenzamide. Solid-state NMR was used to show the disappearance of reactive surface hydride species (M(H)) and to identify newly formed bonded chemical species on the silica surface. DRIFT spectroscopy, elemental analysis, and specific surface-area determinations (BET) of the prepared phases are also reported. PMID- 14556046 TI - FT/ICR-mass spectrometry in nanotechnology: the investigation of metalloid clusters. AB - The particularity of metalloid clusters as a special kind of metal atom cluster is described. For the first time such metalloid clusters are investigated in the gas phase by means of FT/ICR-mass spectrometry, the results of which show that metalloid clusters represent a bridge between the bulk metal and metal compounds that can be found in solution after oxidation of the bulk metal. The metalloid clusters presented herein are [Ga19R6]- (R=C(SiMe3)3), and SiAl14Cp*6 and the precursor Al4Cp*4 (Cp*=eta5-C5Me5). PMID- 14556048 TI - Tetracyclines for treating multidrug-resistant Acinetobacter baumannii ventilator associated pneumonia. AB - OBJECTIVE: To report the use of tetracyclines for the treatment of multidrug resistant Acinetobacter baumannii ventilator-associated pneumonia (VAP). DESIGN: Observational case series. SETTING: . The Presley Regional Trauma Center located within the Regional Medical Center, Memphis, Tennessee, USA. PATIENTS AND PARTICIPANTS: Seven critically ill trauma patients with VAP caused by A. baumannii isolates that were resistant to all antibiotics tested except for doxycycline or minocycline. INTERVENTIONS: Patients were treated with IV doxycycline or minocycline for an average of 13.5 (range 9-20) days. MEASUREMENTS AND RESULTS: Doxycycline or minocycline was successful in six of seven patients. CONCLUSIONS: Doxycycline or minocycline may be effective for treating multidrug resistant A. baumannii VAP. PMID- 14556049 TI - Measurement of diaphragm loading during pressure support ventilation. AB - OBJECTIVE: The diaphragmatic pressure-time product (PTPdi) has been used to quantify the loading and unloading of the diaphragm. The validity of the relationship between PTPdi and diaphragm electrical activity (EMGdi) during pressure-support ventilation (PSV) is unclear. We examined this relationship. DESIGN AND SETTING: Physiological study in a physiology laboratory. SUBJECTS: Six healthy adults. INTERVENTIONS: Spontaneous breathing (SB) and two levels of PSV (6 and 12 cmH(2)O), breathing room air and incremental concentrations of carbon dioxide, sufficient to achieve an EMGdi signal of approximately 200% of baseline value. MEASUREMENTS AND RESULTS: We measured the electrical (EMGdi) and mechanical (PTPdi) activity of the diaphragm using oesophageal electrode and oesophageal and gastric balloon catheters. The relationship between EMGdi and PTPdi during SB was linear in five subjects and curvilinear in one. However, with PSV 12 cmH(2)O we observed that the relationship between EMGdi and PTPdi was 'left shifted'; specifically, for any given level of EMGdi the PTPdi was smaller with PSV 12 cmH(2)O than during SB. However, when PTPdi was converted to power (the product of pressure and flow) the tendency to left shift was largely reversed. CONCLUSIONS: We conclude that when assessing of diaphragm unloading during PSV flow measurements are required. Where flow is constant, PTPdi is a valid measure of diaphragm unloading, but if not these data may be used to make an appropriate correction. PMID- 14556050 TI - Identification of QTLs associated with Fusarium head blight resistance in Zhedar 2 barley. AB - Fusarium head blight (FHB) in barley and wheat, caused by Fusarium graminearum, is a continual problem worldwide. Primarily, FHB reduces yield and quality, and results in the production of the toxin deoxynivalenol (DON), which can affect food safety. Identification of QTLs for FHB severity, DON level and related traits heading-date (HD) and plant-height (HT) with consistent effects across a set of environments, would provide the basis for marker-assisted selection (MAS) and potentially increase the efficiency of selection for resistance. A segregating population of 75 double-haploid lines, developed from the three-way cross Zhedar 2/ND9712//Foster, was used for genome mapping and FHB severity evaluation. A linkage map of 214 RFLP, SSR and AFLP markers was constructed. Phenotypic data were collected in replicated field trials from five environments in two growing seasons. The data were analyzed using MQTL software to detect quantitative trait locus (QTL) x environment (E) interactions. Because of the presence of QTL x E, the MQM procedure in MAPQTL was applied to identify QTLs in single environments. We identified nine QTLs for FHB severity and five for low DON. Many of the disease-related QTLs identified were coincident with FHB QTLs identified in previous studies. Only two of the QTLs identified in this study were consistent across all five environments, and both were Zhedar 2 specific. Five of the FHB QTLs were associated with HD, and two were associated with HT. Regions that appear to be promising candidates for MAS and further genetic analysis include the two FHB QTLs on chromosome 2H and one on 6H, which were also associated with low DON and later heading-date in multiple environments. This study provides a starting point for manipulating Zhedar 2-derived resistance by MAS in barley to develop cultivars that will show effective resistance under disease pressure. PMID- 14556051 TI - Evaluating the potential of barley and wheat microsatellite markers for genetic analysis of Elymus trachycaulus complex species. AB - The potential of barley and wheat microsatellite markers for genetic analysis of Elymus trachycaulus complex species was evaluated. A set of 25 barley and 3 wheat microsatellite markers were tested for their ability to cross-amplify DNA from four accessions of E. trachycaulus and two accessions Pseudoroegneria spicata. Thirteen barley (52%) and two (68%) wheat primer pairs successfully amplified consistent products from both E. trachycaulus and P. spicata species. Four of the 15 successful primer pairs produced visible polymorphisms among the accessions tested. A higher successful rate of cross-species amplification of barley and wheat microsatellite markers in E. trachycaulus and P. spicata was found in this study. These primer pairs are now available for use as markers in genetic analysis of E. trachycaulus complex species. Our results suggest that publicly available wheat and barley microsatellite markers are a valuable resource for the genetic characterization of wild Triticeae species. PMID- 14556052 TI - Tobacco-induced alterations to the Fourier-transform infrared spectrum of serum. AB - Infrared (IR) spectroscopy can distinguish differences in the characteristics of diverse molecules by using infrared radiation to probe chemical bonds. Consequently, alterations to the molecular characteristics of tissues and body fluids that help define specific pathological processes and conditions can be identified by IR spectroscopy. This study analyzed the molecular spectrum of cotinine by IR spectroscopy and determined tobacco-induced alterations to the IR profile of serum to establish whether these alterations can differentiate smokers and nonsmokers. The IR spectra of serum samples obtained from 20 smokers and 25 nonsmokers were captured using a FTS-40 IR spectrometer. Linear discriminant analysis method was used to partition the samples into smoker and nonsmoker groups according to the discriminatory patterns in the data and into a validation set to test the accuracy of the trained algorithm in distinguishing smokers and nonsmokers. Cotinine molecules were shown to exhibit a characteristic IR absorption spectrum. Several differences in the sera spectra of the two groups were observed, including an overall shift in the secondary structure of serum proteins favoring increased beta-sheet content in smokers. The overall accuracy of the training and validation sets was 96.7%, and 82.8%, respectively. The identification of specific absorption peaks for tobacco-induced alterations to the IR molecular profile of serum permits the development of an IR spectroscopy technique that can be used to differentiate smokers from nonsmokers. This further extends the utility of IR spectroscopy as a rapidly emerging tool in the field of molecular biodiagnostics. PMID- 14556054 TI - Primary CMV infections--an important differential diagnosis in outpatients. PMID- 14556053 TI - Unraveling the multiple roles of leptin in inflammation and autoimmunity. AB - Initially described as an antiobesity hormone, leptin has subsequently been shown also to influence hematopoiesis, thermogenesis, reproduction, angiogenesis, and immune homeostasis. Leptin links nutritional status and proinflammatory T helper 1 immune responses, and the decrease in leptin plasma concentration during food deprivation leads to impaired immune function. This review focuses on the multiple roles of leptin in chronic inflammation and autoimmunity and suggests new possible therapeutic implications for leptin modulators. PMID- 14556055 TI - A pooled analysis of telithromycin in the treatment of community-acquired respiratory tract infections in adults. AB - BACKGROUND: Thirteen multinational, Phase III studies were conducted to establish the efficacy of telithromycin 800 mg once daily in the treatment of community acquired respiratory tract infections (RTIs). PATIENTS AND METHODS: Data were analyzed from 4,743 adult patients participating across four indications: community-acquired pneumonia (CAP) of mild to moderate severity, acute exacerbations of chronic bronchitis (AECB), acute maxillary sinusitis (AMS) and tonsillitis/pharyngitis. RESULTS: Treatment with telithromycin for either 5 days (AECB, AMS and tonsillitis/pharyngitis) or 7-10 days (CAP and AMS) provided high rates of clinical and bacteriologic cure (5-day, 87.0% and 86.0%, respectively; 7 to 10-days, 90.3% and 90.5%, respectively) that were equivalent to those of a 10 day course of comparator antibacterials (86.5% and 86.5%, respectively). The clinical efficacy of telithromycin extended to high-risk CAP and AECB patients and to all key respiratory pathogens, including Streptococcus pneumoniae strains resistant to penicillin or erythromycin and atypical/intracellular pathogens. Telithromycin was generally well-tolerated across patient groups. CONCLUSION: These findings support the use of telithromycin as an effective therapy for the treatment of community-acquired RTIs. PMID- 14556056 TI - Primary cytomegalovirus infection in an outpatient setting--laboratory markers and clinical aspects. AB - BACKGROUND: Occasionally, primary cytomegalovirus (CMV) infection may give rise to more or less severe clinical illness in immunocompetent adults. We retrospectively analyzed cases of acute CMV infection in medical outpatients. PATIENTS AND METHODS: Over a 6-year period, we identified 22 patients with a febrile illness and hepatitis suffering from primary CMV infection. This was diagnosed on the basis of a strongly positive CMV IgM antibody test result and/or CMV IgG seroconversion. Clinical features as well as relevant laboratory results were analyzed. We also tested available samples for CMV glycoprotein B-specific antibodies and CMV IgG avidity and analyzed results of Epstein-Barr virus (EBV) specific antibody assays. In addition, current age-specific CMV IgG seroprevalence rates were determined using 9,870 routine patient samples. RESULTS: At presentation, all patients complained of malaise and fever higher than 38 degrees C, and many also complained of cephalgia. Most patients who underwent abdominal ultrasonography had an enlargement of the spleen. Most patients had a relative lymphocytosis but only three had a mild leukocytosis. C reactive protein was only slightly elevated in 13 patients; all 22 patients had elevated levels of alanine aminotransferase (ALT) and lactate dehydrogenase (LDH). Half the patients reported travel to areas outside western Europe, mostly to tropical and subtropical areas, within 3 weeks before onset of illness. Primary CMV infection was confirmed by negative anti-gB antibody test results and the absence of high-avidity CMV antibodies. In contrast, despite past EBV infection demonstrated by positive anti-EBNA-1 results, 15 out of 21 patients tested for EBV markers had positive or nonspecific IgM test results. The overall CMV IgG seroprevalence rate in the routine samples was 64.4%, with marked age dependent increases. CONCLUSION: CMV is a relevant differential diagnosis in feverish illnesses accompanied by hepatitis in otherwise healthy adults, about 40% of whom are CMV-naive. Half our patients seem to have acquired their CMV infection abroad, so that a diagnosis of CMV infection needs to be taken into account in travelers, in addition to infectious illnesses more commonly considered in this context, such as dengue or hepatitis A. For diagnosis, both CMV and EBV antibody studies should be performed and the inclusion of assays able to demonstrate past infection is helpful for achieving a definite diagnosis. PMID- 14556057 TI - Immunization with Haemophilus influenzae (Hib) conjugate vaccine following discharge from hospital. AB - BACKGROUND: This study was undertaken to ascertain Haemophilus influenzae type b (Hib) conjugate vaccination status following discharge from hospital and to determine the time taken to vaccinate with Hib conjugate vaccine following hospital discharge of unvaccinated children. MATERIALS AND METHODS: Population based cross-sectional survey involving children aged 3-59 months admitted to a hospital with laboratory confirmed invasive H. influenzae disease. RESULTS: Of 51 eligible children, 42 (82%) had not received Hib conjugate vaccine prior to admission. No children were immunized prior to discharge. Vaccination following discharge took an average of 110 days (range 3-441 days). More than one quarter (28%) remained unvaccinated following discharge. If these children had been immunized, immunization coverage among this group of children would have increased significantly from 73% to 98% (p = 0.0017). CONCLUSION: In the immediate post-Hib conjugate vaccine period, delayed immunization after discharge was identified. This reduces the individual and community benefits of immunization. PMID- 14556058 TI - Cutaneous anthrax in adults: a review of 25 cases in the eastern Anatolian region of Turkey. AB - BACKGROUND: The clinical features, therapy and outcome of anthrax cases from the Elazig province (the eastern Anatolian region) of Turkey seen in our clinic over an 8-year period were reviewed. PATIENTS AND METHODS: The records of 25 anthrax cases observed in our clinic during the period January 1994 to April 2002 were examined. RESULTS: All cases were cutaneous; 18 (72%) patients exhibited malignant pustules and seven (28%) malignant edema. Three of the patients with a malignant pustule developed anthrax sepsis when admitted to our clinic. All cases were treated with penicillin. One patient who had penicillin allergy was treated with ciprofloxacin. In addition, patients with malignant edema were also treated with systemic corticosteroids (methylprednisolone or dexamethasone). Two patients died due to anthrax sepsis; one case with anthrax sepsis recovered. The mortality rate was 8%. DISCUSSION: Anthrax is still a reality in Turkey. Cutaneous anthrax should be considered in any patient with a painless ulcer with vesicles, edema and a history of exposure to animals or animal products. In our series, penicillin and ciprofloxacin were effective in treatment of anthrax. Our anthrax sepsis case demonstrates that anthrax sepsis is not always fatal if antibiotic treatment is given early after diagnosis. PMID- 14556059 TI - Analysis of prognostic factors in 95 patients with Acinetobacter baumannii bacteremia. AB - BACKGROUND: Because Acinetobacter baumannii bacteremia is a global problem, we were motivated to characterize this disease in Taiwan. PATIENTS AND METHODS: We analyzed findings in 95 patients with documented A. baumannii bacteremia between January 1, 1998 and December 31, 2000 (47 men, 48 women; mean age 58.8 years). RESULTS: The mean length of stay in the hospital was 44.0 days. Clinically, 76 patients had fever and 35 patients developed shock. Fifty patients had respiratory tract infections; 24, urinary tract infections; 11, intra-abdominal infections; three, CNS infections; and two, catheter-related infections. Five patients had primary bacteremia. Empirical antibiotic therapy was initiated at the onset of the clinical signs of infection. Antimicrobial susceptibility test results were variable. 47 patients died and 48 survived; the mortality rate for A. baumannii bacteremia was 45.3% (43/95). CONCLUSION: Physicians should pay attention to this infection because the early identification of high-risk patients could facilitate prophylaxis and potentially reduce associated problems. PMID- 14556060 TI - Spectrum of opportunistic infections and profile of CD4+ counts among AIDS patients in North India. AB - BACKGROUND: As the number of AIDS cases increases in India, information available among clinicians about the prevalence of opportunistic infections (OIs) is scarce. The aim of the present study was to document the characteristic OIs of HIV-infected North Indian patients along with their CD4+ counts. PATIENTS AND METHODS: The study group consisted of subjects with confirmed serodiagnosis of HIV, attending the medical clinics at a tertiary health care center in North India. The CD4+ counts were estimated by FACS Calibur (BD) flow cytometer. Simultaneously, routine microbiology smears, cultures and serology were performed to confirm OI. RESULTS: In this retrospective study of 421 subjects, the predominant OI was tuberculosis (47%, 189 cells/ micro l), followed by parasitic diarrhea (43.5%, 227 cells/ micro l) and oral candidiasis (25.2%, 189 cells/ micro l). CONCLUSION: Tuberculosis was the most frequent OI in the HIV-infected patients studied, and the major mode of transmission of HIV was by sexual route. The median CD4+ counts observed were lower when compared to other studies. PMID- 14556061 TI - Clinical significance of human intestinal spirochetosis--a morphologic approach. AB - Intestinal spirochetosis (IS) is a condition defined morphologically by the presence of spirochetal microorganisms attached to the apical cell membrane of the colonic and rectal epithelium. Intestinal spirochetes comprise a heterogeneous group of bacteria. In humans Brachyspira aalborgi and Brachyspira pilosicoli predominate. Prevalence rates of IS are low where living standards are high, in contrast to poorly developed areas where IS is common. Homosexuals and HIV-infected individuals are at high risk of being colonized. Clinical significance in individual cases has remained unclear up to now. A review of the literature reveals that invasion of spirochetes beyond the surface epithelium is associated with gastrointestinal symptoms which respond to antibiotic treatment (metronidazole), whereas patients lacking this feature are mostly asymptomatic. Homosexual and HIV-positive men are more likely to be symptomatic irrespective of invasion. Rare cases of spirochetemia and multiple organ failure have been reported in critically ill patients. PMID- 14556063 TI - Influenza B pneumonia with Staphylococcus aureus superinfection associated with parvovirus B19 and concomitant agranulocytosis. AB - An 11-year-old patient with anamnestic fever for 3 days and signs of upper respiratory tract infection underwent fulminant Staphylococcus aureus pneumonia with concomitant agranulocytosis. From autopsia influenza B virus and parvovirus B19 were detected by nucleic acid amplification technique (NAT). Specific IgG but no IgM points to preexisting parvovirus B19 infection. Whether in this case agranulocytosis can be interpreted as early manifestation of reactivated parvovirus B19 infection is under discussion. Therefore, parvovirus B19 could have provoked a foudroyant course of influenza B pneumonia which was superinfected with S. aureus. PMID- 14556062 TI - Safety of heart-lung machines during prolonged standby. AB - BACKGROUND: Coronary artery bypass surgery is increasingly performed without an extra-corporeal circuit. A heart-lung machine (HLM) is kept on standby for safety reasons, but rarely used. The tubes of the machines are currently discarded after every operation. Costs and waste would be avoided if HLMs could stay on standby for longer periods of time. We therefore investigated the sterility of intra-tube fluid over time. MATERIALS AND METHODS: Four machines were tested. The tube system was fixed and filled with priming solution. Samples of intra-tube fluid and fluid from the fluid bags were taken after 0, 12, 24, 36, 48 and 72 h. Microbiological cultures were performed by direct inoculation and membrane filtration and incubated up to 14 days. Endotoxin levels were also determined. RESULTS: Bacteria were grown from two samples only and were considered as contaminants. Endotoxin concentrations never exceeded acceptable levels. CONCLUSION: To keep HLMs on standby for 72 h is probably safe. The prolonged use of HLMs will contribute to cost and waste reduction in open heart surgery. PMID- 14556064 TI - Multilevel epidural abscess formation with paraplegia in a healthy 33-year-old man caused by Staphylococcus aureus (MSSA). AB - We report an unusual case of a devastating multilevel pyogenic spondylitis with paraplegia and soft tissue abscess formation in a previously healthy young man. Methicillin susceptible Staphylococcus aureus (MSSA) was identified as causal pathogen. The infection could only be managed after surgical debridement of all spinal manifestations and a prolonged course of antibiotic therapy. It is possible that delayed surgical debridement of all infection sites fostered the course of the disease. PMID- 14556065 TI - Gonococcal tonsillar infection--a case report and literature review. AB - Oral gonococcal infection is an uncommon but well-described manifestation of gonococcal infection, usually described as pharyngitis in the literature. Tonsillitis is much rarer and its role in the clinical presentation in oral gonorrhea is less clear. We describe a case of oral gonorrhea presenting with tonsillitis and a discrete cervical lymphadenopathy and present a review of the literature from 1961 to 2002. Of the 512 reported cases of oral gonococcal infection, only 61 have been described to be tonsillitis. The tonsils were invariably enlarged and infected. A whitish-yellow exudate in the cryptae was described in 12 cases (20.6%). Fever and cervical lymphadenopathy appear to be rather uncommon, since they have been described in only five (8.2%) and six (9.3%) of the 61 patients with tonsillitis, respectively. Gonococcal tonsillitis should be included in the differential diagnosis of tonsillitis in sexually active patients. PMID- 14556066 TI - High level of resistance to levofloxacin of Streptococcus pneumoniae following fluoroquinolone therapy. PMID- 14556067 TI - The relationship between Chlamydia pneumoniae seropositivity and peripheral vascular diseases, acute myocardial infarction and late-onset asthma. PMID- 14556068 TI - The effect of forest clear-cutting on habitat use in Sichuan snub-nosed monkey (Rhinopithecus roxellana) in Shennongjia Nature Reserve, China. AB - The habitat use of two groups of Sichuan snub-nosed monkey ( Rhinopithecus roxellana) was observed, using the focal group method, for 7 months in four seasons from June 2000 to October 2001. The habitats were classed into primary forest and three successional habitats: after clear-cutting: grassland, shrub forest and young forest. The results showed that the large group of monkeys had larger range areas than the small group in the same season. Both groups had larger range sizes in summer or autumn than in winter or spring. They spent most of their time using primary forest and young forest, rarely used shrub forest and did not use grassland. In each season, they used the habitats non-randomly and preferred primary forest. The preference order of habitats for both groups every season was the same: primary forest > young forest > shrub forest >/= grassland. The results suggested that primary forest was high-quality habitat and should be conserved as a top priority. Clear-cutting would cause habitat loss and habitat fragmentation, and should therefore be prohibited. High-quality habitat for the monkeys is difficult to restore from clear-cutting. PMID- 14556071 TI - Uptake of amplifiable fragments of retrotransposon DNA from the human alimentary tract. AB - Few attempts have been made to study the transfer of DNA from ingested food across the intestinal barrier. A low uptake of ingested DNA has been observed in mice, cattle and poultry. There have been no reports on humans so far. Maintenance of species barriers, protection against retrotransposons, optimisation of oral DNA vaccines and the fate of genetically modified foodstuffs are issues where this topic is of importance. We therefore used the high-copy number rabbit retrotransposon RERV-H, and rabbit mitochondrial DNA, to study the transfer of DNA from ingested rabbit meat into the bloodstream of two human volunteers. A quantitative PCR was used to measure RERV-H levels in food and in the blood. Amplification with the primers selected results in the generation of a 250-bp fragment of RERV-H. Transfer across the intestinal epithelium could be demonstrated in both subjects. Levels of the fragment in the bloodstream peaked at 1-3 h after ingestion of the experimental meal. One hour after a meal of rabbit meat containing 10(14) copies of RERV-H DNA, a maximum concentration of 200 copies of RERV-H DNA per ml of peripheral blood was observed, which corresponds to the uptake of approximately 10(6) RERV-H DNA copies in 1 h. RERV-H DNA was detected in both cellular and plasma compartments. Both rabbit retrotransposon and mitochondrial DNA was taken up from the human alimentary tract. The size of the fragments detected is similar to that of SINE retrotransposons (approximately 300 bp). The fate and functionality of alimentary DNA in humans will require further study. PMID- 14556072 TI - Difference in cytoplasmic localization pattern of neutral mucin among lobular endocervical glandular hyperplasia, adenoma malignum, and common adenocarcinoma of the uterine cervix. AB - To investigate whether the cytoplasmic localization pattern of neutral mucin differs between lobular endocervical glandular hyperplasia (LEGH) and cervical adenocarcinoma (CxAd), including minimal-deviation adenocarcinoma (MDA), or adenoma malignum, alcian blue (pH 2.5)/periodic acid-Schiff (AB-PAS) staining was performed to formalin-fixed paraffin-embedded tissue sections of 13 lesions of LEGH and 53 tumors of CxAd, including 6 tumors of MDA. The cytoplasmic localization of neutral mucin was classified as a "whole cytoplasmic pattern," in which neutral mucin filled the cytoplasm entirely, or as an "apical pattern," in which neutral mucin was localized in the subsurface area only. Cytoplasmic neutral mucin patterns were detected in all 13 cases of LEGH and in 19 cases (36%) of CxAd, including five cases of MDA. The localization of neutral mucin was always the whole cytoplasmic pattern in 13 cases of LEGH, but was the apical pattern in these 19 cases of CxAd. The other 34 cases of CxAd, including 1 case of MDA, corresponded to the acid mucin pattern stained purple or blue or no staining pattern by AB-PAS. Among the 53 cases of CxAd, the apical neutral mucin pattern was an indicator of poorer patient prognosis by univariate and multivariate analyses. The examination of cytoplasmic localization of neutral mucin might be applicable, not only to differential diagnosis between LEGH and CxAd, including MDA, but also to estimate clinical aggressiveness of CxAd. PMID- 14556073 TI - Comparison of the effects of phorbol dibutyrate and low-frequency stimulation of synaptic inputs on the excitability of myenteric AH neurons. AB - Low-frequency stimulation of synaptic inputs to after-hyperpolarising (AH) neurons in the guinea-pig small intestine causes sustained increases in excitability that far outlast the stimulus period. This excitation has been called sustained, slow, post-synaptic excitation (SSPE). Intracellular microelectrodes were used to record the effects of the protein kinase C (PKC) stimulant, phorbol dibutyrate (PDBu), and compare these with changes seen during the SSPE, in AH neurons of the small intestine of the guinea-pig. PDBu (1 nM-1 microM) increased excitability, depolarised the membrane and increased input resistance concentration dependently, mimicking the effects of low-frequency stimulation of pre-synaptic inputs. These changes developed slowly after the start of infusion and were only slowly reversible after wash out. PDBu suppressed a late after-hyperpolarising potential (AHP) that depends on Ca2+ entry via voltage-gated Ca2+ channels during the action potential. The effects of PDBu (10 nM) on the late AHP were indistinguishable from those observed during the SSPE. PDBu, at a concentration that inhibited the AHP, had no effect on the action potential half-width or the slope of its first repolarisation phase (the early phase of repolarisation is slowed by the Ca2+ influx of the action potential). Thus PDBu inhibited K+ channel opening underlying the late AHP, but did not suppress Ca2+ entry during the action potential. The hyperpolarisation-activated cation current (Ih) in intrinsic primary afferent neurons (IPANs) was not affected by PDBu. We conclude that PDBu mimics the sustained excitation caused by low-frequency stimulation of synaptic inputs to IPANs by closing IK channels responsible for the AHP or restricting their opening by Ca2+ and by reducing the current carried by K+ channels that are active at rest. IK channels, the opening of which results in the AHP, have consensus sites for PKC and are likely targets for phosphorylation during the SSPE. PMID- 14556074 TI - Mitochondria and release at hippocampal synapses. AB - Mitochondria are present in some, but not all presynaptic terminals in the hippocampus. Mitochondria are capable of sequestering and storing large amounts of calcium, but it is unclear whether they influence release probability at these synapses. Using FM dye imaging techniques and confocal microscopy, we have examined the relationship between mitochondrial presence/absence and presynaptic vesicle release from rat hippocampal neurones in primary dissociated culture at room temperature. Following staining with the mitochondrial dye mitotracker green, we were able to resolve putative individual mitochondria associated with neuronal processes. The majority of mitochondria were positionally stable, although some exhibited periods of rapid motility (up to 0.4 microm/s) interspersed with periods of immobility. Co-staining with mitotracker green and the synaptic vesicle dye FM 4-64 indicated that 180 of 506 (36%) synapses were devoid of mitochondria. A comparison of vesicular release in response to stimulation at 1 Hz and at 10 Hz revealed no differences in release properties between synapses with and without mitochondria. PMID- 14556075 TI - RhoA induction by functional overload and nandrolone decanoate administration in rat skeletal muscle. AB - The regulation of skeletal muscle regeneration and growth in response to functional overload is a coordinated interaction of mechanical and endocrine signaling pathways. This study's purpose was to determine if RhoA expression and activity in rat plantaris muscle was induced by functional overload with or without anabolic steroid administration. Male Sprague Dawley (125 g) rats were subjected to bilateral ablation of the gastrocnemius muscle for 3 and 21 days and treated with nandrolone decanoate (ND, 6 mg/kg b.w.) or sesame seed oil injections. Western blot analysis revealed that RhoA protein expression was induced 2.1-fold by overload and 1.9-fold by ND at 3 days. RhoA protein remained elevated by overload after 21 days (3.8-fold). In addition, RhoA protein expression in C2C12 myotubes was induced after 18 h of 1% (1.8-fold) or 2% (2.2 fold) chronic radial stretch. Competitive RT PCR revealed that RhoA mRNA concentration increased 1.9-fold with ND, 2.9-fold with overload, and 11.8-fold with overload and ND administration when compared to sham at 3 days, indicating pre-translational control of RhoA by ND and a synergism between ND and overload to up-regulate RhoA mRNA. The ratio of RhoA protein associated with the muscle membrane fraction, an indicator of RhoA activity, increased 3.7-fold after 3 days of overload compared to controls. Although ND with overload (3.8-fold) produced a larger induction of RhoA protein than overload alone, the ratio of RhoA protein associated with the membrane fraction was not altered by ND treatment at 3 days. In conclusion, RhoA is an integrator of both mechanical and growth factor signaling whose expression and activity are increased by the combination of anabolic steroid and functional overload treatments in rat plantaris muscle. PMID- 14556076 TI - Oxygen uptake kinetics: old and recent lessons from experiments on isolated muscle in situ. AB - The various mechanisms responsible for ATP resynthesis include phosphocreatine (PCr) hydrolysis, anaerobic glycolysis and oxidative phosphorylation. Among these, the latter represents the most important mechanism of energy provision. However, oxidative phosphorylation is characterized by a lower maximal power and a slow attainment of a steady state in response to increased metabolic demand. The rate of adjustment of oxidative metabolism during metabolic transitions, which can be evaluated on the basis of the analysis of O2 uptake (VO2) kinetics, has implications for exercise tolerance and muscle fatigue. Analysis of VO2 kinetics represents a valid tool for the functional evaluation of healthy subjects, athletes and patients. Over the last 35 years experiments conducted on isolated muscle preparations in situ have allowed us to gain insights into several key aspects of skeletal muscle VO2 kinetics. Their main limiting factor resides in an intrinsic slowness of intracellular oxidative metabolism when adjusting to augmented metabolic needs. The rate of adjustment of oxidative phosphorylation in mitochondria can be functionally related to PCr hydrolysis occurring in the cytoplasm. PMID- 14556078 TI - Increase of KL-6 in sera of uveitis patients with sarcoidosis. AB - BACKGROUND: KL-6 is a human glycoprotein secreted by type II alveolar cells in lung, and its serum levels increase in pneumonia of various causes. KL-6 is a member of the MUC-1 family, which is expressed in lung, cornea, and conjunctiva. The purpose of the present study was to investigate the clinical usefulness of quantifying serum KL-6 levels for diagnosing sarcoidosis in patients with uveitis. METHODS: Sera were obtained from 24 uveitis patients diagnosed with sarcoidosis, 37 uveitis patients with other etiologies, and 138 healthy control subjects. Serum concentration of KL-6 was determined by a human KL-6 electrochemiluminescence immunoassay. RESULTS: The average level of KL-6 in the sera of uveitis patients with sarcoidosis was 387 U/ml. This was significantly higher than in healthy subjects and uveitis patients with other etiologies. The KL-6 measurements identified 45.8% of sarcoidosis-positive patients. When the KL 6 results were combined with serum angiotensin-converting enzyme (ACE) concentrations, 87.5% of sarcoidosis patients were identified, compared to 66.7% using ACE results alone. The combined measurement identified 10.8% of the non sarcoid patients and 0.72% of healthy subjects as positive (false positive). CONCLUSION: Combined measurements of serum KL-6 and ACE may be useful as a screening for sarcoidosis in uveitis patients. PMID- 14556077 TI - Long-term results after photodynamic therapy with verteporfin for choroidal neovascularizations secondary to inflammatory chorioretinal diseases. AB - BACKGROUND: To evaluate the safety and the long-term effect on visual acuity of photodynamic therapy (PDT) for sub- and juxtafoveal choroidal neovascularizations (CNV) secondary to inflammatory conditions. METHODS: In a prospective pilot study, 19 patients with CNV due to inflammatory conditions underwent PDT treatment with verteporfin with standard parameters. Regular follow-up was carried out every 3 months with ETDRS visual acuity measurement and fluorescein angiography. The initial diagnosis included punctate inner choroidopathy (PIC) ( n=7), presumed ocular histoplasmosis syndrome (POHS) (6), multifocal choroiditis with panuveitis (MCP) (2) and other inflammatory conditions (4). RESULTS: All patients had a minimum follow-up of one year. After a mean follow-up of 22.1 months, the mean change in visual acuity was +1.63 ETDRS lines. At their last visit, all eyes showed a disappearance of pretreatment leakage in fluorescein angiography and 63.2% (12/19) eyes had an improvement of 2 or more lines, while 26.3% (5/19) remained stable (+/-1 line) and 10.5% (2/19) lost 2 or more lines. A mean of 2.0 (1-5) treatments was performed. No clinically relevant side effects or complications were observed. CONCLUSIONS: PDT is a safe and effective treatment option for CNV secondary to inflammatory conditions. The results are better than for CNV secondary to AMD. For juxtafoveal CNV, the results are similar to those of subfoveal CNV with no additional safety concerns. Based on this observation, we consider PDT as treatment of choice for subfoveal CNV secondary to inflammatory chorioretinal diseases and for selected cases with juxtafoveal CNV. PMID- 14556079 TI - Functional thrombomodulin deficiency causes enhanced thrombus growth in a murine model of carotid artery thrombosis. AB - Thrombomodulin (TM) bound thrombin initiates the protein C anticoagulant pathway and defects in TM result in enhanced coagulation. Recent studies suggest a role for TM in arterial vascular disease. In order to corroborate this association we studied arterial thrombus formation in mice with a functional TM defect. We used mice homozygous for a (404)Glu-to-Pro mutation in the TM gene (TM(pro/pro)) and compared these with wildtype littermates in a model of FeCl(3) induced carotid artery thrombosis. Time-to-occlusion (TTO) was assessed by arterial blood flow measurement, using a Doppler flow probe. Complete occlusion occurred in 8/10 (80%) TM(pro/pro) mice and in 3/11 (27%) littermate controls. Mean time to occlusion (TTO) [+/- SE] was 767 +/- 196 s in the F2-TM(pro/pro) mice, versus 1507 +/- 159 s in controls (p = 0.007, Mann Whitney U test). Histology and immunostaining for tissue factor did not reveal any differences in thrombus morphology or thrombogenicity between the two groups. These data confirm and extend the finding that a functional deficiency in TM results in enhanced thrombus formation in a murine model of carotid artery thrombosis and support a role for TM defects in arterial thrombotic disease. PMID- 14556080 TI - HMG-CoA reductase inhibition reduces the proinflammatory activation of human vascular smooth muscle cells by the terminal complement factor C5b-9. AB - The terminal complement complex C5b-9 is known to participate in inflammatory processes including atherosclerosis. Inflammation appears to be a direct consequence of C5b-9-mediated cell stimulation. 3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors may exert anti-inflammatory effects on vascular cells independent of lowering plasma cholesterol. Thus, we studied activation of vascular smooth muscle cells (VSMCs) by C5b-9 focusing on whether inhibition of the HMG-CoA reductase can reduce the proinflammatory effects of C5b 9.C5b-9 in sublytic concentrations increased the proliferation of human VSMCs and induced a time-dependent activation of the mitogen-activated protein (MAP) kinase extracellular signal-regulated kinase (ERK). Proliferation and ERK1/2 activation could be inhibited by the specific ERK inhibitor PD98059. HMG-CoA inhibition with cerivastatin-reduced VSMC proliferation and C5b-9-induced ERK1/2 activation. Cerivastatin also reduced the C5b-9-induced synthesis of the proinflammatory interleukin-6 (IL-6). Furthermore, C5b-9 induced activation of the transcription factors activator protein- 1 (AP-1) and nuclear factor-kappaB (NF-kappaB), which could be inhibited by pretreatment of VSMCs with cerivastatin. L-mevalonate and geranylgeranylpyrophosphate reversed the inhibitory effects of cerivastatin. The present study in VSMCs shows that cerivastatin inhibits IL-6 synthesis and cell proliferation induced by the terminal complement complex C5b-9. This may be an important mechanism contributing to the beneficial effects of HMG-CoA reductase inhibitors beyond lowering of plasma cholesterol. PMID- 14556081 TI - A hyperpolarization activated inward current (If) is present in infant ventricular myocytes. AB - I(f) was shown to be present in adult human atrial and ventricular myocytes but data obtained from infant myocytes are lacking. We have studied I(f) in isolated ventricular myocytes from children undergoing surgical correction of tetralogy of Fallot (TOF; n = 5; mean age: 15.3 months). All recordings were made with the patch clamp technique in the whole cell mode at a temperature of 36-37 degrees C. A modified Tyrode solution containing 25 mM KCl was used to amplify I(f). Considering I(f) to be present when its current density at -120 mV was greater than 0.5 pA/pF, I(f) could be found in 28 out of 32 myocytes (88%). The mean current density was -2.01 +/- 0.3 pA/pF (mean +/- S.E.M.). First current activation occurred at -70 mV and I(f) could be reversibly inhibited by superfusing the myocytes with CsCl (2 mM). Half maximal activation (V(1/2)) of I(f) was at -80.3 +/- 1.0 mV (n = 28). Beta-adrenergic receptor stimulation with isoproterenol (1 microM) caused an acceleration of current activation and a shift of V(1/2) by 7.88 +/- 1.8 mV (n = 10) to less negative potentials. This study provides first evidence that the hyperpolarization-activated pacemaker current I(f) is present in infant human ventricular myocytes. Our results suggest that I(f) in ventricle of infants suffering from TOF has similar properties as I(f) in adult ventricle. PMID- 14556082 TI - Regional and frequency-dependent changes in action potentials and transient outward K+ currents in ventricular myocytes from J-2-K cardiomyopathic hamsters. AB - OBJECTIVES: Although lengthening of action potential duration (APD) and decreased transient outward K+ currents (I(to)) have been observed in ventricular myocytes from cardiomyopathic hamsters, epi- and endo-cardial differences in I(to) and their roles in frequency-dependent changes in APD have not been clarified. METHODS: The patch-clamp technique of whole-cell configuration was used to record membrane potentials and currents in epicardial and endocardial myocytes of the J 2 hamster germline without (control) and with cardiomyopathy (CM). RESULTS: In control, APD in endocardial myocytes was longer than that in epicardial myocytes at 0.1 Hz. APD significantly lengthened with increased frequencies of stimulation from 0.1 to 6.0 Hz in both groups with the longer APD in endocardial myocytes. In CM, APD lengthened in epicardial myocytes exceeding the endocardial APD without a frequency-dependent prolongation. Pretreatment with 4 mM 4-aminopyridine completely abolished the frequency-dependent changes and abolished APD differences between epicardial and endocardial myocytes, and between control and CM hamsters. The transient outward K+ current (I(to)) significantly decreased in epicardial myocytes from CM hamsters compared with that of control (17.5 +/- 1.5 pA/pF in control vs. 9.5 +/- 2.5 pA/pF in CM at +60 mV) with altered recovery from inactivation, without changes in the endocardial I(to). Moreover, the inward rectifier K+ current decreased in epicardial myocytes from CM hamsters and the L type Ca2+ current reduced in both regions from CM compared to control. CONCLUSION: Results indicate that differences in APD between epi- and endocardial myocytes in CM hamsters are mainly caused by a decreased current density and altered recovery from inactivation of I(to) in epicardial myocytes. PMID- 14556083 TI - Cardiac-directed expression of adenylyl cyclase and heart rate regulation. AB - Mice with cardiac-directed overexpression of AC(VI) show increased cardiac responsiveness to beta-adrenergic receptor stimulation but regulation of heart rate is unknown. Telemetry was used to test the hypothesis that mice overexpressing cardiac adenylyl cyclase type VI (AC(VI)) would have normal heart rate regulation. Mice overexpressing cardiac AC(VI) were generated using the alphaMHC promoter and studied 10 days after implantation of telemetry devices. Cardiac transgene AC(VI) presence and expression was verified using PCR, RT-PCR and immunoblotting. Ambulatory heart rates were assessed using time and frequency domain analysis over two 24 hour light-dark cycles. Heart rates then were assessed following pharmacological blockade. Time domain analyses showed ambulatory heart rates were unchanged (AC(VI): 597 +/- 15 (SEM) bpm, Control: 595 +/- 12 bpm; p = 0.92). Circadian heart rate variability was preserved and not different from control mice (ANOVA, p = 0.52). Frequency domain analysis of heart rate variability also was unchanged. No difference in heart rate response to pharmacological autonomic blockade was found (intrinsic heart rate: AC(VI) 622 +/ 17 bpm, control 616 +/- 16 bpm, p = 0.79). In conclusion, mice overexpressing cardiac AC(VI) have normal conscious ambulatory heart rates and normal heart rate variability. Overexpression of cardiac AC(VI) does not result in altered heart rate regulation in contrast to cardiac overexpression of other elements of the beta-adrenergic signaling pathway. PMID- 14556084 TI - Putative role of epithelial sodium channels (ENaC) in the afferent limb of cardio renal reflexes in rats. AB - Recent studies suggest a role of ion channels of the DEG/ENaC family for mechanosensation in different species and in baroreceptor reflex control in rats. We tested the hypothesis that ENaC within the cardiac sensory network are mandatory for mechanosensation. Experiments were performed in male Sprague-Dawley rats, isolated nodose ganglion cells with cardiac afferents and isolated vagus nerves. Epicardial delivery of the amiloride analogue benzamil intended to specifically inhibit ENaC presumably located on cardiac sensory afferents indeed blunted the mechanosensitive (i. e., sympathoinhibition by intravenous volume loading [-32% and -42% in treated groups vs. -67% in controls; n = 7 each; p < 0.05]) as well as-though to a lesser extent-the 5-HT(3)-mediated chemosensitive cardiorenal reflex in vivo in a dose-dependent manner. Using patch clamp technique, however, it turned out that neither amiloride nor benzamil influenced mechanically induced currents in ganglion nodosum cells in vitro, stimulated by hypoosmotic stress. The unspecific stretch activated ion channel blocker gadolinium completely abolished mechanically induced currents, indicating respective cells were mechanosensitive. In isolated vagus nerves benzamil impaired action potentials obtained by electrical stimulation (C-spike amplitude [-33%]; latency [+12%]; n = 8; p < 0.05). Our findings at least cast doubt on ENaC exclusively playing a specific role as mechanotransducers within the cardiac sensory network. Other ion channels might be involved. Furthermore the observed findings in vivo could also be due to unspecific disturbance of afferent signal conduction. PMID- 14556085 TI - Effect of ghrelin and synthetic growth hormone secretagogues in normal and ischemic rat heart. AB - Receptors for growth hormone secretagogues have been identified in cardiac tissue, but their functional role is unknown. We have investigated the effect of different growth hormone secretagogues on contractile performance and on the susceptibility to ischemic injury, in isolated working rat hearts. In particular, we tested the endogenous secretagogue ghrelin and the synthetic secretagogues hexarelin and MK-0677. Under aerobic conditions, none of these substances produced any significant hemodynamic effects. In hearts subjected to 30 minutes of ischemia followed by 120 minutes of reperfusion, the synthetic peptidyl secretagogue hexarelin (1 microM) significantly reduced infarct size, as estimated on the basis of triphenyltetrazolium chloride staining, while the non peptidyl secretagogue MK-0677 was ineffective. The endogenous peptidyl secretagogue ghrelin (20 nM) was also protective, while desacylated ghrelin, which is devoid of biological effects, did not modify ischemic injury. The protection provided by hexarelin was partly abolished by the protein kinase C inhibitor chelerythrine. We conclude that ghrelin and hexarelin have a specific cardioprotective effect, which is independent of growth hormone secretion, and might be related to protein kinase C activation. PMID- 14556086 TI - Coronary vascular dysfunction associated with direct current shock injury. AB - OBJECTIVE: To determine if cardiac injury following DC shocks includes impairment of coronary vascular reactivity. METHODS: 36 dogs (18-32 kg) were anesthetized and a thoracotomy was performed. Either antioxidant enzymes, superoxide dismutase (SOD, 15,000 U/kg) plus catalase (55,000 U/kg) or the NO synthase inhibitor N(G) nitro-L-arginine (L-NNA, 5 mg/kg) was administered IV prior to sham (no shocks) or DC shock treatment, and the results were compared to dogs which did not receive SOD/catalase or L-NNA. In sham dogs, electrodes cradled the heart, but no shocks were delivered. In shock dogs, three 20 Joule DC shocks were delivered to the epicardium using hand-held paddles. Other dogs were allowed a 3-hour recovery period after the shocks. Epicardial microvessels and conduit rings were studied in vitro. Antagonists were not added to the bath of the study vessel. Internal diameter was measured in microvessels after constriction with endothelin. Tension of conduit arteries was measured after constriction with PGF(2alpha). Responses to acetylcholine (Ach, 10(-10)-10(-4) M), bradykinin (10(-14)-10(-6) M), the calcium ionophore A23187 (A23187, 10(-12)-10(-4) M) or nitroprusside (SNP, 10( 10)-10(-4) M) were measured. RESULTS: Bradykinin, A23187 and SNP dependent dilation was not different between vessels from sham and shocked animals. Dilation to Ach was attenuated in vessels from shocked dogs. Superoxide production probably contributed to the impaired dilation to Ach since treatment with SOD/catalase improved dilation. Treatment with L-NNA also improved vascular function after DC shock. CONCLUSION: DC shocks cause endothelial dysfunction, as demonstrated by impaired dilation to acetylcholine, in both canine coronary microvascular and conduit arteries. Since pretreatment with either SOD/catalase or L-NNA protects against this damage, a free radical mechanism, possibly involving eNOS, may contribute to endothelial dysfunction.DC shocks for cardioversion and defibrillation cause myocardial injury that may be free radical mediated. PMID- 14556087 TI - Coronary blood flow regulation in the prediabetic metabolic syndrome. AB - This study tested whether the pre-diabetic metabolic syndrome impairs coronary blood flow control sufficiently to alter the balance between coronary blood flow and myocardial metabolism. Experiments were conducted in dogs instrumented with catheters in the aorta, coronary sinus, and left ventricle and with flow transducers around the circumflex coronary artery and aorta. Coronary blood flow, myocardial oxygen consumption (MVO(2)), cardiac output, aortic pressure, left ventricular pressure and heart rate were measured at rest and during treadmill exercise in normal, control and high fat fed dogs. High fat feeding for approximately six weeks increased body weight 15%, increased aortic blood pressure 10%, and induced insulin resistance. Fasting plasma insulin levels were increased 2.4-fold while plasma glucose concentration was unchanged relative to controls (5.0 +/- 0.3 mM). The cardiac index increased with exercise but was not altered by high fat feeding. The metabolic syndrome reduced the slope of the relationship between coronary blood flow and MVO(2) ( P < 0.0001) and decreased coronary venous PO(2) at a given level of MVO(2) ( P < 0.05). These findings indicate that the metabolic syndrome impairs the balance between myocardial oxygen delivery and metabolism by tonically vasoconstricting the coronary circulation. PMID- 14556091 TI - [Strengthening the capacity of reference in response to violence in society and within the family]. AB - Results of longitudinal studies in developmental psychology have shown the importance of protective factors especially within high risk samples for adaptive development as they serve as puffers against many risks. An outstanding protective factor is a secure parent-child attachment relationship. This article outlines the different ways how attachment relationships can serve as protective mechanisms and especially how disorganized attachment relationships are leading to maladaptation and hostile attitudes and behaviors. In doing so it underscores the special importance of attachment experiences in early infancy. The concept of attachment relationship is described as opposed to the bonding concept with regard to its implications for practice. One of the first is to intervene early. In the final part the evidence based early intervention program "STEEP" is described as an application of attachment research on a poverty sample and as a response to violence. PMID- 14556092 TI - [Standards in the perinatal medicine--Doppler sonography in the gestation]. PMID- 14556093 TI - [The oxytocin signal cascade during premature labor]. AB - Since 30 years the rate of preterm deliveries of 7% remained unchanged. This is due to a lack of understanding of the underlying pathomechanisms of preterm labor. Oxytocin (OT) as well as its receptor (OTR) play a key role in the process of (preterm) labor as part of a paracrine system that regulates myometrial contractility. Binding of OT to its corresponding receptor, OTR, leads to activation of actin-myosin interactions and therewith myometrial contractions as well as production of intrauterine prostaglandins (PGE(2), PGF(2 alpha)) mainly in decidua and myometrium. Oxytocin expression increases significantly at time of parturition, as does the expression of its receptor. Both can be influenced by diverse cellular substrates. The focus of our research group is based on the exploration of the influence of cytokines on OTR signaling. PMID- 14556094 TI - [Biopsychosocial predictors of preterm labor and preterm delivery? Results of a prospective study]. AB - RESEARCH QUESTION: In addition to medical, job related, sociodemographic risk factors, and health related behavior, topics that traditionally have been less in the focus of attention in this context, such as biographic data, coping with stress, personality variables, pregnancy related attitudes, fears, and the social network were examined. PATIENTS AND METHODS: 589 women between 16(th) and 22(nd) week of pregnancy were examined using a questionnaire that was designed for the study. This resulted in 508 women pregnant with a single child, whose pregnancy and delivery were examined based on their medical records. Factor analysis and main component analysis with subsequent varimax rotation resulted in factors that were subject to a proof of reliability. Statistical analysis was based on logistic regression. RESULTS: 129 (27.7%) of women displayed signs/indices of an imminent preterm delivery, 29% (5.8%) of whom later actually had a preterm delivery before the end of the 37(th) week of pregnancy. Pregnant women who were in treatment for an imminent preterm delivery appear to have been subject to higher social stress as compared to those, who later later actually gave birth before completing the 37(th) week of pregnancy. Lack of a female network and lack of emotional understanding from the partner are correlated to both of these complications. Specifically, actual preterm delivery appears to be significantly influenced by partner relationship. Another significant predicting variable for imminent and actual preterm delivery appears to be a history of gynecological problems. A distinct risk factor for delivery before completion of the 37(th) week of pregnancy was a history of colpitis. In addition, pronounced anxieties in respect to the pregnancy, and low general anxiety were significant predicting variables for delivery before completion of the 37(th) week of pregnancy. CONCLUSIONS: Partner relationship, female networks, psychosomatic reactivity in terms of diseases/disorders of the reproductive organs, and anxieties appear to be worthwhile targets in the prevention of preterm delivery. PMID- 14556095 TI - [Measurement of volume flow by "colour velocity imaging (CVI)": technique of measurement in the intrahepatic fetal umbilical vein]. AB - "Colour Velocity Imaging (CVI)" permits acquisition of ultrasound Doppler data for quantitative analysis. CVI can be used to study the fetal circulation. In a pilot study we attempted measuring umbilical venous flow. Data acquisition is simple and fast. In the current implementation, data is sent to a work-station and analysed off-line. There is significant "intra-observer-variability" that could be attributable to true physiological variation or measurement inaccuracies. Measurement of fetal umbilical volume flow by CVI is a new and simple technique. Before its clinical use assumptions about the vessel, flow, variability and normal values must be verified in the fetus. PMID- 14556096 TI - [Association between ovarian stimulation regime and protein metabolism of patients undergoing IVF/ICSI therapy]. AB - The aim of this study was to determine the relationship between the ovarian stimulation protocol (with HMG or FSH) after down-regulation with GnRH anologa and protein (total protein and albumin) as well as bilirubin and urea in serum. Furthermore, it was intended to find out the effect of these parameters on IVF/ICSI outcome. 50 patients were included in this study. All patients underwent controlled ovarian hyperstimulation for assisted reproduction therapy either with FSH (Gonal-F) or HMG (Menogon). Ovulation induction was induced by human chorionic gonadotrophine (HCG, Predalon) 10 000 IU i. m. The protein concentration (total protein, albumin) as well as bilirubin and urea concentrations were determined before down-regulation with GnRHa, at the beginning of ovarian stimulation with FSH or HMG, on the day of ovulation induction with HCG, during oocyte retrieval and fourteen days after embryo transfer. The age, body mass index and etiology of infertility showed no significant difference between patients stimulated with HMG or FSH. Total protein and albumin concentration decreased significantly (p=0.001) from 77.45 +/- 5.90 g/L and 47.02 +/- 3.41 g/L to 74.60 +/- 4.6 g/L and 45.04 +/- 2.39 g/L, respectively at the time of oocyte retrieval. Whereas, no significant change with bilirubin and urea concentration was observed. However, the mean concentration of total protein, albumin, bilirubin of patients who become pregnant was higher of those who did not. In conclusion, this study shows that total protein and albumin concentration in plasma decreased during the follicular phase significantly with the application of exogenous gonadotrophins and steroid hormones in comparison to the value before down-regulation. The mean value of total protein, albumin, bilirubin of patients who become pregnant was higher (but not significant) of those who did not. However, the high individual variation in the present results shows that these parameters are not useful as a predictor of IVF/ICSI outcome. PMID- 14556097 TI - [Placenta praevia percreta--a serious complication after previous cesarean section]. AB - A 36-year-old G4P1 presented at 20 weeks gestation with vaginal bleeding. Her obstetrical history was significant for two first-trimester spontaneous abortions requiring curettage and a cesarean section. On admission placenta praevia was suspected by ultrasound. A placenta percreta was suspected by ultrasound follow up at 30 weeks. At 33 weeks she underwent cesarean section because of serious vaginal bleeding. The profound blood loss with consecutive coagulopathy required an emergency hysterectomy and multiple blood transfusions. Placenta percreta is a rare but dramatic complication after previous cesarean section. This should be kept in mind as the rate of elective cesarean sections is rising continuously. PMID- 14556098 TI - [Workflow improvement and efficiency gain with near total digitalization of a radiology department]. AB - PURPOSE: To determine the temporal changes of the workflow caused by digitalization of the radiology department after installation of digital luminescence-radiography (DLR), a radiology information system (RIS) and picture archiving and communication system (PACS) at the Missionsarztliche Klinik in April 2000. MATERIALS AND METHODS: In a comparative study, a workflow analysis by manual registration of different work steps was performed before (1999) and after (2001) digitalization of a radiology department. RESULTS: The digitalization shortened the examination time for patients from a mean of 8 min to 5 min. The time the patient is absent from the emergency room did not change. Reporting radiographic examinations including comparison with previous studies begins earlier from a mean of 2 h 37 min to 17 min. Using PACS, 85.9 % of all cases could be interpreted on the day of the examination (without PACS 41.2 %) and 87.2 % of the reports were completed the day after the examination (without PACS 64.5 %). No time differences were found between reading conventional studies on the monitor or as soft-copy. CONCLUSION: Compared to conventional film-screen systems, complete digitalization of a radiology department is time saving at nearly all steps of the workflow, with expected positive effects on the workflow quality of the entire hospital. PMID- 14556099 TI - [Normal and abnormal water diffusion in the brain]. AB - Diffusion magnetic resonance imaging (MRI) has become an important tool in the radiologic diagnosis of diseases of the brain as it measures molecular motion of water that characterizes the microstructure of tissues. Its most important clinical use to date is the early detection of cerebral ischemia by revealing the ischemic injury shortly after vessel occlusion and simultaneously providing therapy-relevant information on the tissue at risk. Furthermore, diffusion MRI is diagnostically promising in other diseases of the brain and is thus increasingly becoming part of routine clinical protocols in the diagnosis of tumors, inflammation, trauma, demyelination, dysmyelination and neurodegeneration. Although abnormalities of diffusion are generally not pathognomonic, diffusion MRI affords information about tissue changes for specific disorders that complements information obtained with standard MR techniques and frequently shows pathology earlier. In addition, diffusion MRI can be applied to plan, guide and follow-up biopsies or resective surgery. Particularly diffusion tensor imaging (DTI), which displays the orientation of white matter fibers, holds promise for improved surgical planning. Moreover, DTI can be used to detect changes in connectivity between functional brain areas. Therefore, DTI is highly relevant not only in advancing the knowledge of white matter diseases but also in stimulating research on normal brain development and brain aging. PMID- 14556100 TI - Coronary MR imaging using free-breathing 3D steady-state free precession with radial k-space sampling. AB - PURPOSE: To investigate the potential of free-breathing 3D steady-state free precession (SSFP) imaging with radial k-space sampling for coronary MR angiography (MRA), coronary projection MR-angiography and coronary vessel wall imaging. MATERIALS AND METHODS: A navigator-gated free-breathing T2-prepared 3D SSFP sequence (TR = 6.1 ms, TE = 3.0 ms, flip angle = 120 degrees, field-of-view = 360 mm(2)) with radial k-space sampling (384 radials) was implemented for coronary MRA. For projection coronary MRA, this sequence was combined with a 2D selective aortic spin tagging pulse. Coronary vessel wall imaging was performed using a high-resolution inversion-recovery black-blood 3D radial SSFP sequence (384 radials, TR = 5.3 ms, TE = 2.7 ms, flip angle = 55 degrees, reconstructed resolution 0.35 x 0.35 x 1.2 mm(3)) and a local re-inversion pulse. Six healthy volunteers (two for each sequence) were investigated. Motion artifact level was assessed by two radiologists. RESULTS: In coronary MRA, the coronary lumen was displayed with a high signal and high contrast to the surrounding lumen. Projection coronary MRA demonstrated selective visualization of the coronary lumen while surrounding tissue was almost completely suppressed. In coronary vessel wall imaging, the vessel wall was displayed with a high signal when compared to the blood pool and the surrounding tissue. No visible motion artifacts were seen. CONCLUSION: 3D radial SSFP imaging enables coronary MRA, coronary projection MRA and coronary vessel wall imaging with a low motion artifact level. PMID- 14556101 TI - [Evaluation of the contribution of the importance of neuroimaging for the diagnostics of dementias--comparison to the psychological diagnostics]. AB - OBJECTIVE: While psychology is accepted as a necessary component of the dementia diagnostics, the extended clinical diagnostics with neuroimaging is differently estimated. The goal of the study is the quantification of the individual contribution of the two different methods. METHODS: Of 100 patients the diagnosis of entrance, the neurological, the psychological, and the final clinical diagnosis were documented. For both imaging and psychology the sensitivity, specificity, and the positive predictive value were computed. The diagnostic of each method was determined from the change of the final in relation to the initial clinical diagnosis. The neuroradiological investigation took place with MRI, the psychological examination used both usual power and special speed tests. RESULTS: The extended clinical diagnostics led for 26 % of the patients to the change of the clinical diagnosis. Imaging and psychology supplied different own but supplementing contributions. In the case of annihilation imaging contributed with 73.3 %, psychology with 54.1 % to the diagnosis of a neurodegenerative dementia, whereas the contributions to the diagnosis of a vascular dementia were 83.3 % and 70.8 %, respectively. However psychology diagnosed and quantified the dementia. The contribution of neuroimaging consisted in the differential diagnosis of the dementias. Organic causes of symptomatic dementias and vascular encephalopathy without dementia but with consequences for a secondary prophylaxis were additional information also. CONCLUSION: Psychology improves the diagnostic accuracy of dementias. Neuroimaging improves the differential diagnosis of dementias and supplies additional clinically relevant findings. In the qualified diagnostics and differential diagnostics of the dementias both methods are indispensable. PMID- 14556102 TI - [Quantitative analysis of contrast enhanced MRI of the inferior alveolar nerve in inflammatory changes of the mandible]. AB - PURPOSE: To evaluate the role of contrast enhanced MRI in quantifying signal changes of the inferior alveolar nerve following inflammatory changes of the mandible. MATERIAL AND METHODS: 30 patients with inflammatory changes of the mandible underwent MRI of the face. Both sides of the mandible, the affected as well as the unaffected healthy side were evaluated retrospectively. Regions of interest were placed at 5 defined places on both sides to assess signal intensity before and after intravenous application of paramagnetic contrast agent. The results of the measurements were compared between the healthy and the affected side (t-test, p < 0.05) and correlated with clinical findings (t-test, p < 0.05). RESULTS: All patients with hypesthesia of the inferior alveolar nerve in areas of the lips or chin (n = 4) showed a marked increase in signal intensity from 35 % to 83 % distal to the inflammatory process. Patients without sensitivity disorders showed less increase in signal intensity with a maximum of 51 % distal to the inflammatory process. In nearly all patients no contrast enhancement was observed distal to the first molar on the unaffected side. CONCLUSIONS: Quantitative analysis of contrast enhanced MRI of the neurovascular bundle is able to show pathologic mandibular lesions. In case of inflammatory changes of the mandible the neurovascular bundle is able to show pathologic mandibular lesions. In case of inflammatory changes of the mandible, the neurovascular bundle shows a significant increase in signal intensity distal to the lesion compared to the unaffected healthy side of the mandible. PMID- 14556103 TI - [Left ventricular functional parameters using ECG-gated multidetector spiral CT in comparison with invasive ventriculography]. AB - PURPOSE: To evaluate the accuracy of left ventricular function using a multidetector CT (MDCT) with retrospective ECG-gating and to compare the results with conventional ventriculography. MATERIALS AND METHODS: In 26 patients, retrospectively ECG-gated MDCT of the heart (Volume Zoom, Siemens, Germany) as well as conventional coronary angiography including ventriculography was performed to exclude or follow coronary artery disease. CT examination (120 KV, 400 mAs) was carried out with 4 x 1 mm collimation (500 ms gantry rotation time). For CT angiography, 150 ml of contrast media was injected intravenously at a flow rate of 4 ml/s. All data sets of the functional cardiac parameters were reconstructed in end-systolic and end-diastolic phase. End-systolic volume (ESV), end-diastolic volume (EDV) and ejection fraction (EF) were determined from multiplanar reformations orthogonally through the cardiac short axis and analyzed using special evaluation software (ARGUS, Siemens). The results were compared with ESV, EDV and EF obtained from invasive ventriculography. RESULTS: In all cases, a sufficient quality of the MDCT images was achieved. EDV (150.1 +/- 16.2 ml MDCT vs. 138.7 +/- 16.9 ml ventriculography; mean difference 11.4 +/- 12.7 ml; r = 0.51) had an acceptable correlation to conventional ventriculography, and ESV (58.1 +/- 14.6 ml vs. 50.2 +/- 13.4 ml; mean difference 7.9 +/- 8.8 ml; r = 0.81) and EF (60.9 +/- 13.6 % vs. 64.9 +/- 12.7 %; mean difference 4.0 +/- 6.2 %; r = 0.79) showed a good correlation. In comparison with invasive ventriculography, MDCT tended to overestimate significantly EDV (p = 0.008) and ESV (p = 0.003) and to underestimate EF (p = 0.001). CONCLUSION: MDCT of the heart with retrospective ECG-gating enables efficient estimation of left ventricular function, providing important additional information of non-invasive cardiac imaging using MDCT. However, EDV and EVS were significantly overestimated and EF was underestimated in MDCT compared to ventriculography. PMID- 14556104 TI - [Multislice TrueFISP-MR imaging for identifying stress-induced myocardial functional disturbances in coronary heart disease]. AB - PURPOSE: This study assesses the left ventricular function using a new multislice cine sequence and determines the diagnostic accuracy of stress-induced wall motion abnormalities in patients with coronary artery disease (CAD). MATERIALS AND METHODS: 15 patients (mean age 57.7 years) with angiographically proven CAD were examined on a 1.5 T whole body system (Magnetom Sonata, Siemens, Erlangen) at rest and during dipyridamole-induced (0.56 mg/kg body weight) stress. Left ventricular function was determined using a multislice (steady-state) sequence (TR 2.3 ms, TE 1.15 ms, slice thickness 10 mm, temporal resolution 77 ms) as well as a standard single-slice true FISP 2D sequence (TR 3.2 ms, TE 1.6 ms, slice thickness 5 mm, temporal resolution 45 ms) as reference. RESULTS: Both cine sequences provide high sensitivity and excellent correlation (r = 0.95) with angiographic findings for the detection of regional wall motion abnormalities. However, the measurement of functional parameters yielded significant differences. End-systolic left ventricular volumes (ESV) were systematically overestimated in the multislice images (mean 78 ml, + 5.8 %) compared with the reference single-slice images (mean 74 ml) (p < 0.05). This resulted in underestimation of the ejection fraction with multislice images (mean 40 %, - 11.3 %) compared with single-slice images (mean 46 %) (p < 0.05). CONCLUSION: The multislice sequence results in a substantial reduction of imaging time and breath hold periods necessary to cover the left ventricle for functional assessment. The multislice sequence yields adequate images, especially for qualitative determination of wall motion abnormalities. Due to the reduced spatial and temporal resolution of the multi-slice sequence, however, some uncertainty concerning the functional parameters has to be taken into account. PMID- 14556105 TI - [Enhanced MR-angiography (CE-MRA) of the supraaortic arteries: dose reduction with a contrast agent of high relaxivity (Gd-BOPTA)]. AB - PURPOSE: Reducing the amount of contrast medium in contrast-enhanced MR angiography (CE-MRA) of the cervical vessels could lead to considerable cost reduction. This study investigates whether the amount of contrast medium possessing high relaxivity can be reduced without loss of diagnostic information. MATERIALS AND METHODS: In a prospective study the supraaortic vessels of 40 patients were examined on a 1.5 T MR-system using 10 or 20 ml Gd-BOPTA (MultiHance). The examinations were evaluated by measurement of the signal increase in the common carotid arteries in 38 patients. Furthermore, CE-MRA was rated by two independent observers, who were blinded to the amount of contrast medium and degree of stenosis. Rating criteria were quality of arterial contrast and image impression. RESULTS: Signal increase in the carotid artery was identical for 10 and 20 ml Gd-BOPTA in 12 patients with optimal bolus timing (10 ml: 454 +/- 58; 20 ml: 458 +/- 63). Signal intensities were significantly lower for 10 ml when bolus timing was suboptimal (281 +/- 80 vs 353 +/- 65; p < 0.02). The quality of the CE-MRA using 10 and 20 ml Gd- BOPTA was rated as identical by the observers, and the amount of contrast medium used could not be deduced from the images. CONCLUSION: Image quality and signal values of the carotid arteries did not differ in CE-MRA performed with either 10 ml or 20 ml Gd-BOPTA (MultiHance) as long as the examination is adequate. Thus, cost reduction can be achieved with a reduced standard dose of 10 ml Gd-BOPTA without loss of diagnostic information. PMID- 14556106 TI - [Use of SPIO-enhanced T1- and T2-weighted images for the differentiation of liver lesions: an ROC analysis]. AB - PURPOSE: The superparamagnetic iron oxide (SPIO) Resovist is a contrast media with shortening of both T(1) and T(2) relaxation time. This study evaluates the impact of SPIO-enhanced T(1)- and T(2)-weighted images for the differentiation of liver lesions. MATERIALS AND METHODS: SPIO-enhanced MRI examinations (1.5 T, Symphony Quantum) of 61 patients were analyzed. Thirty-seven patients had malignant liver lesions (metastases n = 32, HCC n = 5) proven by biopsy or laparotomy, 11 patients had benign liver lesions (FNH n = 2, hemangiomas n = 4, benign cysts n = 5, normal liver on laparoscopy n = 13). After unenhanced T(1)- and T (2)-weighted imaging, a bolus injection of 1.4 ml SPIO (Resovist) was given, followed by T(1)-weighted imaging at 20 s, 60 s, and 5 min and T(2) weighted imaging at 10 min post injection. A score from 1 (benign) to 5 (malignant) was used by three blinded radiologist for the ROC analysis of the unenhanced T(1)-/T(2)-weighted images (set 1) and of the combinations of unenhanced T(1)/T(2)-weighted and SPIO T(1)-weighted images (set 2), unenhanced T(1)/T(2) and SPIO T(2)-w images (set 3) and all images (set 4). RESULTS: The accuracy of plain MRI (set 1: 56 %) was increased by SPIO-enhanced T(1)-weighted images (set 2: 81 %) and SPIO-enhanced T(2)-weighted images (set 3: 90 %). Best results were obtained using unenhanced T(1)-weighted, unenhanced T(2)-weighted and both SPIO T(1)-weighted and T(2)-weighted images (set 4: 93%). The accuracy of predicting histopathologic diagnosis was 91%. CONCLUSION: For the differentiation of liver lesions, SPIO-enhanced T(2)-weighted images had a greater impact on the accuracy of MRI than T (1)-weighted images, but SPIO enhanced T(1)-weighted images provided additional information in some patients and should not be deleted. PMID- 14556107 TI - [Detection of liver lesions with gadolinium-enhanced VIBE sequence in comparison with SPIO-enhanced MRI]. AB - PURPOSE: Detection of malignant liver lesions with gadolinium-enhanced volumetric interpolated breath-hold examination (VIBE) in comparison with SPIO-enhanced MRT (four different SPIO-enhanced T2w sequences) and histological and imaging follow up in non-cirrhotic livers at 1.5 T. PATIENTS AND METHODS: Twenty-two patients with suspected focal liver lesions prospectively underwent a gadolinium-enhanced 3D VIBE. Four T2w sequences (HASTE sequence, fast spin-echo-sequence with and without fat-saturation, FLASH 2D gradient echo-sequence) after administration of superparamagnetic iron oxide (SPIO) served as gold standard combined with histological work-up in 17 patients and histological and imaging follow-up in five patients. The image quality was evaluated and the detectability of intrahepatic lesions was rated by the alternative free-response receiver operating characteristic (AFROC) analysis. In addition, the contrast-to-noise ratio was compared. RESULTS: Altogether 49 malignant and 35 benign liver lesions were found. Concerning the image quality, VIBE turned out to be of slightly poorer image quality than the SPIO-enhanced examination with HASTE sequence (4.95 vs. 5.0). The fast spin-echo-sequence without fat-saturation demonstrated the highest contrast-to-noise ratio. All sequences showed a comparable certainty in detecting lesion (area under the curve 0.68-0.73) and identifying malignant liver lesions. CONCLUSION: Despite the small number of patients, VIBE seems to be a comparable, inexpensive and fast method in diagnosing malignant liver lesions. PMID- 14556108 TI - [CT-colonography with the 16-slice CT for the diagnostic evaluation of colorectal neoplasms and inflammatory colon diseases]. AB - PURPOSE: Comparison of the performance of virtual and conventional colonoscopy for the detection of colorectal polyps and inflammatory colon diseases using a 16 slice spiral CT scanner. Furthermore, presentation of the first experiences with a new three-dimensional reconstruction mode ("colon-dissection") that dissects the colonic wall like a pathologic-anatomic preparation to increase the sensitivity. MATERIALS AND METHODS: Forty patients were studied using a 16-slice spiral CT (Lightspeed 16, General Electric Medical Systems, Milwaukee, Wisconsin 53201, USA). The examination was performed after standard oral preparation for colonoscopy. The colonic distension was achieved with room air and intravenous butylscopolamine. Images were obtained in supine and prone position using a detector configuration of 16 x 0.625 mm, pitch 1.7, rotation time 0.5 s, 160 mAs and 120 kV. Axial reconstruction with a slice thickness of 0.625 mm. The CT data were assessed by two radiologists on an Advantage Workstation (Volume Analysis 2, USA) using a software with the capabilities of axial, multiplanar and volume rendering, virtual endoscopy, and colon dissection. Conventional colonoscopy was used to determine the sensitivity. RESULTS: A total of 30 polyps were found in 8 patients and a carcinoma was detected in two patients. Colonography identified 4 polyps with a diameter of 10 mm or more, 6 polyps with a diameter of 5 mm to 9.9 mm, 11 polyps with a diameter of 3 to 4.9 mm and 9 polyps with a diameter of 3.0 mm or less. There were two false negative findings (one polyp of 3 mm and one of 4 mm had been overseen) and two false positive findings for polyps (polyps of 4 mm and 6 mm). The sensitivity and specificity for the detection of colonic polyps were 93% and 94% with the "colonic-dissection" mode, 87 % and 94 % with the "virtual-endoscopy" mode and 63 % and 97 % with multi-planar reconstruction, respectively. Depending on the diameter of the colonic polyps, the "colon dissection" mode ("virtual-coloscopy") had a sensitivity and specificity of 100 % and 100 % for polyps with a diameter over 5.0 mm, 91 % and 82 % for polyps with a diameter from 3.0 to 4.9 mm and 89 % and 78 % for polyps with a diameter under 3.0 mm, respectively. Inflammatory colon diseases presented as thickening of the colon wall over 5 mm. CONCLUSIONS: Virtual colonoscopy with 16-slice spiral CT allows accurate detection of colonic polyps, including small polyps below a diameter of 3 mm. In comparison with the 2D- and 3D-"virtual-endoscopic" reconstruction, the 3D-reconstruction software "colon-dissection" achieves the highest sensitivity for the detection of colonic masses. Therefore, the combination of a 16-slice spiral CT and the "colon-dissection" reconstruction software provides a high resolution in the z-axis for detecting colonic masses and polyps down to a diameter of less than 2 mm, with a sensitivity of about 90 %. This sensitivity is much higher than the sensitivity achievable with 4-slice spiral CT and without "colon-dissection" mode. A thickened colon wall over 5 mm indicates inflammatory colon disease. PMID- 14556109 TI - [Endovascular therapy of abdominal aortic aneurysm: results of a mid-term follow up]. AB - PURPOSE: Prospective study to evaluate clinical results and complications of endovascular abdominal aortic aneurysm treatment in a mid-term follow-up. MATERIALS AND METHODS: A total of 122 patients (9 females, 113 males, average age 70.9 +/- 7.9 years) with abdominal aortic aneurysms were treated with stent grafts (53 Vanguard or Stentor endografts, 69 Talent endografts). Group I consisted of 40 patients who had all aortic tributaries of the aneurysm sac occluded prior to endovascular grafting, either spontaneously by parietal thrombosis or by selective coil embolization of the respective ostia preserving collateral circulation distal to the vessel occlusion. Group II consisted of 82 patients and included all cases without or with incomplete coil embolization with at least one patent vessel. Stent grafting was performed in general anesthesia in the first 21 patients, followed by peridural anesthesia in 15 cases, and local anesthesia with conscious sedation in 86 cases. The results were evaluated with Spiral-CT, MRI and radiographs of the endovascular graft, with follow-up examinations obtained at 3, 6, 12 months, and every year. RESULTS: Implantation was successfully completed in all cases without primary conversion surgery, laparotomy, or any significant complication. Mean follow-up was 29 +/- 21 months (maximum 84 months). The 30-day mortality was 0.8 % due to a myocardial infarction 3 days after discharge from the hospital. A total of 47 re interventions were performed in 29 patients (23.8 %), with 35 re-interventions in 18 cases with Vanguard or Stentor endografts and 12 re-interventions in 11 patients with Talent endografts. 23 percutaneous re-interventions included distal graft extension (n = 11), Wallstents for kinking and limb stenosis (n = 3), and secondary coil embolization of collateral vessels (n = 9). 24 surgical re interventions included proximal graft extension (n = 6), new endovascular grafts (n = 3), surgical clipping of lumbar and mesenteric artery branches for type-II endoleaks following ineffective secondary coil embolization (n = 1), and femorofemoral crossover bypasses (n = 4). A total of 10 secondary conversion operations were performed because of damage to the membrane (n = 4; 3 Vanguard endografts, 1 Talent endograft), significant caudal migrations (n = 5; 4 Vanguard endografts, 1 Talent endograft) associated with type-I endoleaks (n = 2), limb occlusion (n = 1), disconnection of graft components (n = 1), and significant endoluminal thrombus deposits (n = 1). One patient, who was followed for 82 months, suffered from a significant endoleak for 10 months with increasing aneurysm diameter but he refused surgery. He was admitted with aneurysm perforation and was successfully operated with aortic graft replacement. Compared to group II, the incidence and size of endoleaks was reduced in group I (incidence 19.2 % versus 29.9 %, p < 0.05). Group I demonstrated significantly better aneurysm shrinkage at 36 months follow-up (Delta sagittal diameter - 11.1 +/- 8.4 mm versus - 4.9 +/- 6.2 mm, p < 0.05). CONCLUSION: In selected patients, endovascular aneurysm treatment is an effective alternative to open surgery. It is safely performed in local anesthesia with low mortality rate and a low number of acute complications. Intermediate follow-up revealed re-interventions in around one quarer of all patients, especially when Vanguard or Stentor endografts were implanted. Primary coil embolization of all aortic branches prior to endovascular grafting improves clinical outcome. Insufficient proximal fixation and its consecutive complications remains a major problem of this method. PMID- 14556110 TI - [Contrast-enhanced ultrasound with Optison in percutaneous thermoablation of liver tumors]. AB - PURPOSE: To detect vascularization of liver tumors by ultrasound enhanced with Optison and to determine whether increasing necrosis during a percutaneous thermal ablation can be visualized by this method. MATERIALS AND METHODS: Twenty two patients with non-resectable malignant liver tumors (9 patients with HCC, 13 patients with metastases) and a total number of 34 lesions underwent percutaneous radiofrequency thermoablation using a needle applicator perfused with a 0.9 % NaCl solution (Electrotome HiTT 106, Berchtold). The tumor size ranged from 2 to 8 cm with an average size of 3.6 cm. While intermittent energy was supplied during thermoablation, multi-slice CT (Volume-Zoom, Siemens) and ultrasound were obtained. Ultrasound was performed with a multi-frequency transducer (3 - 7 MHz, LOGIQ 700, GE) and 0.5 to 1 ml of Optison as contrast agent. If necessary, the injection of the contrast agent was repeated after about 30 min. The mechanical index (MI) was set low (0.2 - 0.3) to prevent bubble destruction. Ultrasound power Doppler (PD), contrast harmonic imaging (CHI) and coded harmonic angiography (CHA) as subtraction mode of harmonic imaging were applied intermittently. RESULTS: Only 21 of the 34 lesions showed increased intratumoral perfusion when the conventional B-Mode with PD was used. Similar to spiral CT, the tumor vascularization in the early arterial phase was only reliably visible with CHA after application of contrast agent. CHI with PD was best to detect increasing hypoperfusion of the tumors. When the energy supply was continued, hyperechoic border zones became visible around the central hypoechoic defects. The contrast between remaining tumor and surrounding liver tissue improved in CHI for up to 30 min after the bolus injection of contrast medium, considerably facilitating the evaluation of perfusion. Metal artifacts and the limited amount of contrast agent that can be safely administered interfered with monitoring the thermal ablation with spiral-CT. After an administered energy of maximal 100.000 watts, no more tumor vascularization was seen in 28 of 34 cases. Follow-up spiral CT showed a complete necrosis in these cases. CONCLUSION: Evaluating perfusion with contrast-enhanced ultrasound may be helpful in monitoring the evolving necrosis during thermoablation of liver tumors. The more reliable assessment of the tumor necrosis enables a more targeted therapy. PMID- 14556111 TI - [Postsurgical computed tomography of uncemented hip arthroplasty]. AB - PURPOSE: Presentation of the value of postsurgical computed tomography (CT) to diagnose loosening of uncemented femoral stems. MATERIALS AND METHODS: Incremental CT and spiral CT were performed on six femora with implanted uncemented stems after the entire femora were embedded in polymethylmethacrylate. The femora were subsequently sectioned (thickness 1 mm, separation 8 mm) and the medial and lateral contact areas of the prosthetic stem compared with the CT data. RESULTS: The CT showed a contact of femoral stem and cortical bone between 0.4 mm (3.4 %) and 4.8 mm (47.1 %) and the section specimens between 0.9 mm (8.7 %) and 3.4 mm (36.7 %). No correlation was found between the results (r = 0.61), since the individual differences were up to 24 % in almost all sections. CONCLUSIONS: Neither single-slice nor two-slice CT is capable of demonstrating the direct bone-endoprosthesis contact. Multidetector row CT (MDCT) is conceivably more accurate to measure the cortical contact of the femoral stem. PMID- 14556113 TI - [Measurement of the AT and the CCD angle of macerated cadaver femora: a comparative study between CT and MRI measurements]. AB - PURPOSE: To compare measurements of the real AT (femoral antetorsion) and CCD (caput-collum-diaphysis) angle by computed tomography (CT) and magnetic resonance imaging (MRI) of hip specimen using direct measurements as reference standard, and to show that measurement by MRI can replace CT measurements and may help avoiding X-ray exposition. MATERIALS AND METHODS: CT and MRI measurements were obtained on 25 in water-arrested macerated human femora. Postprocessing was done by 4 independent readers on a workstation using a dedicated 3D-software. Direct measurements of the real AT and the CCD angle were used as reference standard. The analysis included Student's t test for paired values, interobserver variability using intra-class correlation coefficients (ICC), maximum and middle divergence of the angles, and Bland-Altman plots. RESULTS: For determining AT and the CCD angle with CT and MRI, good correlation was found between the 4 readers and with measurements using the reference standard. ICCs were 0.97 and 0.90 for measuring AT and CCD angle with CT, and 0.95 and 0.71 for measurements with MRI, respectively. Mean divergence between CT measurements and those of the reference standard was 0.8 degrees for AT and 0.7 degrees for the CCD-angle. Mean divergence between MRI measurements and those of the reference standard was 0.3 degrees for AT and -0.4 degrees for the CCD-angle. Mean divergences between CT and MRI measurements of AT and CCD-angle were 0.5 degrees. Neither systematic errors nor dependences on the qualitative size of the reference data were evident in the divergences of measurements. CONCLUSION: Measurements of the real AT and CCD angle by CT and MR imaging revealed a good correlation with direct measurements of the femoral specimen and consequently can be recommended for clinical use. MRI measurements can replace CT measurements, avoiding X-ray exposure especially in young patients undergoing preoperative evaluation for hip dysplasia. PMID- 14556114 TI - [Abscess forming portal vein thrombosis: successful percutaneous catheter drainage]. PMID- 14556112 TI - [Diagnostic efficacy of gadodiamide-enhanced dynamic examination technique in comparison to conventional magnet resonance tomography in exacerbated chronic osteomyelitis]. AB - PURPOSE: To evaluate the diagnostic effectiveness of dynamic gadodiamide-enhanced MRI in discriminating reparative tissue formation from florid inflammation in patients with chronic osteomyelitis. MATERIALS AND METHODS: Subjects with chronic osteomyelitis and clinically suspected exacerbation were consecutively enrolled into a prospective trial. Following conventional contrast-enhanced MRI scanning, a dynamic contrast-enhanced SE sequence was carried out in all patients (acquisition interval 58 s, 11 repetitions with a total acquisition time of 11 minutes). The relative increase of the time-dependent signal intensity was assessed in representative circular regions of interest placed over bone marrow and surrounding soft tissues. As diagnostic reference standard, histologic and intraoperative findings were obtained independently from the result of the index test. The MRI studies were evaluated by two radiologists who were blinded to the findings of the reference test. RESULTS: 30 of 51 patients were subsequently scheduled for surgery (mean age 42.6 +/- 12.7 years, 27 males). 19 cases had a florid granulocytic infection, with 6 cases showing histologic signs of putrid liquefaction. Another 11 patients had chronic-granulating, fibrosing osteomyelitis. The course of the signal-time curves differed significantly between the different pathologic processes (multiple measures ANOVA, p < 0,001) in both bone marrow and soft tissue. The areas under the receiver operating characteristics (ROC) curves of the dynamic sequence as a performance measure to distinguish between acute and chronic infection were 0.73 (95 % confidence interval 0.41 - 1.00) and 0.82 (95 % confidence interval 0.59 - 1.00), respectively. Although no overall difference was found in diagnostic efficacy between conventional and dynamic MRI, the dynamic technique provided important additional information for equivocal conventional results. CONCLUSION: Dynamic contrast-enhanced MRI scanning is a safe and potentially valuable tool to discriminate between florid and chronic inflammation in documented osteomyelitis. PMID- 14556115 TI - [Limb-girdle muscular dystrophy in MRI]. PMID- 14556116 TI - [Left ventricular myxoma--a rarity]. PMID- 14556117 TI - [Phytoestrogens]. AB - The soy-isoflavones genistein and daidzein and the flaxseed-lignans secoisolariciresinol and matairesinol belong to the group of phytoestrogens. Epidemiological data suggest that phytoestrogens have a preventive effect against various estrogen-related diseases/symptoms such as breast cancer, menopausal symptoms, cardiovascular diseases, and osteoporosis. To prove these assumptions, available controlled clinical trials have been critically reviewed. Especially soy-isoflavones have been extensively studied. There is no scientific evidence for an effect of phytoestrogens on menopausal symptoms and risk factors of breast cancer. However, isoflavones-containing soy protein can lower total cholesterol, LDL cholesterol, and triglyceride serum levels. The strongest evidence exists for a preventive effect of soy isoflavones on postmenopausal bone loss of the lumbar spine. Distinct effects on estrogen-related diseases can be explained at least in part by the different affinity of isoflavones to estrogen receptors alpha and beta and the distinct tissue distribution of these receptors. PMID- 14556118 TI - [Peri- and postmenopausal changes--body awareness, sexuality, and self- image of the middle-aged and older woman]. AB - INTRODUCTION, METHODS: For the first time a study has been made, in a German urban population, of sexuality, body awareness and self-image of women between 50 and 82 years. A special anonymous questionnaire was designed by female gynaecologists and psychologists for this cross-sectional study in Munich. It focuses on body awareness, self imagery, sexual activity and psychological and physiological sexual feeling in their biographic context. 102 women took part. RESULTS: 53.9% are sexually active and 74.5% think of themselves as "a knowing woman", "middle aged" or "in the best years". Self perception of the body as being attractive correlates highly significantly with the psychological well being. There is no correlation between women's psychological well-being and the perception of their general beauty. Women who find their body attractive think of themselves as more sensual than those, who do not find their body attractive. General and bodily perception of beauty are negatively correlated to the body mass index. Here, 61.8% of the women think they are overweight, but only 41.2 % have a body mass index over 25. 12.9% were sexually abused, and another 8.8% do not want to answer this question. There is a strong correlation between fear and sexual abuse. Only about 50% of the women speak about their sexuality and, of them, only 9.8% speak with their gynaeclologist. CONCLUSION: Sexuality is very important for the middle-aged and older woman, their body perception is as sensitive as in younger women and is of enormous importance for their psychological well-being. A Sexual history should be obligatory for older women. Fear of the examination can be a hint towards sexual abuse and should be taken seriously. All therapeutic interventions such as medication or operations should be discussed, and the patient's sexual wishes and expectations have to be taken into consideration. PMID- 14556119 TI - [Pregnancy in young girls]. AB - More and increasingly young adolescents are sexually active and pay less or no attention to contraception. The pregnancy rate among adolescents has been definitely increasing in recent years and especially among the youngest. This entails a large proportion of medicosomatic and also psychosocial risks such as premature labour and delivery as well as social disadvantages. Moreover, abortions have greatly increased. e.g. by 20% in 2001. Among the causes besides earlier maturity we may assume inadequate sex education falling short of cognitive capacity and emotional needs as well as difficult social problems with deficiencies in tenderness requirements and poor prospects of vocational training and professional career 8 examples, case reports). The overall situation pre programmes medium-term and long-term consequences such as deficient school education and training, financial dependence and excessive financial and emotional strain on mother and child. A considerable gap in this regard can be filled by medical health promotion in step with physical and mental development, as well as by primary and secondary prevention in respect of information deficits, within the framework of a sound relationship of trust and reliability between the physician and the (female) patient. PMID- 14556120 TI - [New perspectives in intrauterine surveillance with the fetal magnetoencephalogram]. AB - PURPOSE: Despite intensive research and surveillance up to now one has failed to reduce cerebral handicaps in newborn. Fetal heart rate tracing (CTG) and Doppler have reduced the number of subpartal severe asphyxia and fetal death. But, 90% of cerebral damage is a result of antepartal problems. Thus only 10% can be avoided by intensive surveillance during labor. Detection of antenatal cerebral injury is a rare case and its impact on later fetal life can only be estimated. Insight in fetal neuronal function is not possible. Factors and time pattern determining fetal cerebral injury are thus not known. This publication explains a new system with whom one might be able to get more insight in cerebral wellbeing during the fetal intrauterine life. METHODS AND RESULTS: A new diagnostic approach is set up by recording fetal magnet encephalographic signals (fMEG) thus offering the opportunity to detect fetal brain function. An array which was especially designed to fit to the pregnant body consists of 151 sensors which are able to record the fMEG. Clinical testing is performed in the moment at the UAMS in Little Rock, Arkansas in Cooperation with the Institutes for Medical Psychology and the Frauenklinik in Tiibingen. First results and arising questions are published. CONCLUSION: With this new system a deeper insight into the fetal neuronal development and fetal wellbeing during pregnancy might be achieved thus reforming the fetal surveillance in the 21st century. PMID- 14556121 TI - [Hypothyroidism in dizygotic premature twins due to excessive prepartal vaginal iodine application]. AB - In dizygotic premature twins delivered by Caesarean section after prolonged efforts to effect premature birth we found markedly enhanced thyroid stimulating hormone levels. The mother had been subjected to local polyvidone iodine therapy for seven weeks with PVP suppositories to prevent vaginal infections. Control checks revealed persistently elevated TSH levels, whereas T3/T4 levels were always reduced. Greatly pronounced iodine excretion was seen in the urine of both the babies. This raised the suspicion of iodine-induced hypothyroidism in both children. Substitution with 1-thyroxine was initiated and a withdrawal trial performed after 12 weeks. The patients remained euthyroid after the treatment had been discontinued. Due to possible transfer of iodide to the fetus associated with the risk of iodine-induced hypothyroidism, it is recommended to abstain from vaginal application of iodine during pregnancy. PMID- 14556122 TI - Finding nerves is not simple. PMID- 14556123 TI - "He said, she said, NSAIDs". PMID- 14556124 TI - You're not as clean as you think! The role of asepsis in reducing infectious complications related to regional anesthesia. PMID- 14556125 TI - Painful paresthesiae are infrequent during brachial plexus localization using low current peripheral nerve stimulation. AB - BACKGROUND: Considerable controversy exists over the relationship of paresthesia to nerve stimulation. The purpose of this study was to determine the frequency with which patients report paresthesia at the point that an acceptable motor response is obtained to low-intensity current electrical stimulation. METHODS: Low-intensity current nerve stimulation (0.6 mA, 200 microseconds, 2 Hz) was used to identify the brachial plexus in 64 consecutive patients having shoulder or arm surgery with an interscalene block. During nerve localization and while maintaining a motor response (0.20 mA-0.40 mA), the patients were queried regarding any radiating sensation or pain (paresthesia) in the shoulder or extremity on the side of the blockade. Sensory distribution of the block, motor strength of the arm muscles, and adequacy of anesthesia were used to assess the extent of blockade. RESULTS: Ninety-five percent of patients had satisfactory surgical anesthesia. None of the patients spontaneously reported having a paresthesia during nerve stimulation. However, on careful questioning, half of the patients (55%) reported electrical paresthesia, defined as dull tingling sensation traveling down to their hands and coinciding with the motor response. In addition, most patients (71%) spontaneously reported having a mild, radiating paresthesia on initial injection of local anesthetic. CONCLUSIONS: Painful paresthesiae should be infrequent when a low-stimulating current is used to identify the neural components of the brachial plexus and when the block needle is advanced slowly. Low-current intensity nerve stimulation can be used to achieve successful interscalene block with minimal discomfort to the patient. PMID- 14556126 TI - Relationship between evoked motor response and sensory paresthesia in interscalene brachial plexus block. AB - BACKGROUND AND OBJECTIVES: This study sought to define the relationship between a paresthesia and a motor response (MR) to electrical nerve stimulation using a peripheral nerve stimulator (PNS) during interscalene block. We sought to determine if at a low amperage (< or =1.0 mA) a MR would precede a paresthesia. METHODS: Twenty-two interscalene blocks were performed using insulated needles and a PNS. A MR was obtained at 0.5 mA and then the PNS was turned off. The needle was further advanced until a paresthesia was elicited. The PNS was again turned on, the needle held immobile, and the amperage increased in 0.1 mA increments up to 0.5 mA, or an MR obtained, whichever occurred first. If no MR was obtained, the needle was withdrawn at 0.5 mA in the same direction as it entered until MR was again observed. RESULTS: A MR was obtained at 0.5 mA in all the patients. After the PNS was turned off and the needle further advanced, a paresthesia was elicited in 21 patients. When the PNS was turned on again, a MR was produced at 0 to 0.5 mA in 13 patients. In a subset of 8 patients without a second MR to stimulation up to 0.5 mA, the needle was withdrawn at that amperage. A MR was subsequently obtained during withdrawal in each patient in this subset. CONCLUSIONS: MR preceded paresthesia in every patient. The most likely explanation for this observation is that MR can be achieved at a small distance from the nerve, whereas elicitation of mechanical paresthesia requires either nerve contact or more intimate location of the needle's tip relative to the nerve. Another possible explanation is that motor fibers are located in a more superficial position and are therefore encountered first. Motor and sensory responses are separate and discrete phenomena. PMID- 14556127 TI - Is skin disinfection with 10% povidone iodine sufficient to prevent epidural needle and catheter contamination? AB - BACKGROUND AND OBJECTIVES: Epidural space infection is a potential complication of epidural catheter placement. In this study, we investigated the incidence of epidural needle and catheter contamination after skin surface disinfection with 10% povidone-iodine (PI). METHODS: Sixty seven patients having surgery under epidural anesthesia were enrolled in this prospective study. After preparation with 10% PI, skin swab cultures were taken from the site of catheter insertion. Epidural needles were cultured immediately after epidural catheters were placed. Catheters were removed at 48 hours and 2 to 3 cm of the distal tips were cultured as well. RESULTS: Fifty-six skin swabs, 52 epidural needles, and 48 catheters were cultured. Although only 3.5% (2) colonization was observed on skin surface cultures, 34.6% (18) of the epidural needles and 45.8% (22) of the catheters were colonized. No systemic or local infection was observed. CONCLUSIONS: Our results suggest that despite skin surface disinfection with PI, there is still significant risk for contamination of needles and catheters during epidural catheterization. PMID- 14556128 TI - A comparison of levobupivacaine 0.5% and racemic bupivacaine 0.5% for extradural anesthesia for caesarean section. AB - BACKGROUND AND OBJECTIVES: Bupivacaine is available as a racemic mixture of its enantiomers, dextrobupivacaine and levobupivacaine. Both in vitro and in vivo studies show that dextrobupivacaine has a greater inherent central nervous system (CNS) and cardiovascular toxicity than levobupivacaine. Clinical studies show levobupivacaine to have similar local anesthetic potency to the racemate. The aim of this study was to investigate the clinical efficacy and safety of levobupivacaine compared with racemic bupivacaine for extradural anesthesia. METHODS: We studied 62 patients undergoing elective caesarean delivery who received 25 mL of either levobupivacaine 0.5% or bupivacaine 0.5%, extradurally, in a randomized, double-blind study. RESULTS: The 2 groups were similar in terms of time to block suitable for surgery, segmental spread, and duration of sensory block. However, lower-limb motor block was significantly longer in the levobupivacaine group but of significantly less intensity. CONCLUSIONS: Levobupivacaine produces an extradural block that is similar to bupivacaine, and is a suitable local anesthetic agent for caesarean delivery. PMID- 14556129 TI - Hemodynamic and blockade effects of high/low epinephrine doses during axillary brachial plexus blockade with lidocaine 1.5%: A randomized, double-blinded study. AB - BACKGROUND AND OBJECTIVES: Although epinephrine commonly is added to local anesthetics for regional anesthesia, rarely it may cause undesirable hemodynamic side effects. This study compared the hemodynamic and blockade effects of 25 and 200 microg epinephrine during axillary brachial plexus blockade with lidocaine 1.5%. METHODS: Sixty American Society of Anesthesiologist classification I or II patients were divided randomly into 3 groups. Patients in group 1 received 5 mL of saline containing 25 microg epinephrine and then 35 mL of 1.5% lidocaine; patients in group 2 received 5 mL of saline alone and then 200 microg of epinephrine mixed with 35 mL of 1.5% lidocaine; patients in group 3 received 5 mL of saline alone and then 35 mL of 1.5% lidocaine. Hemodynamic data were measured for 1 to 10 minutes at 1-minute intervals after axillary injection. The duration time of motor and sensory block was recorded. RESULTS: Complete anesthesia was achieved in 85% of patients in groups 1 and 3 and 90% in group 2. Motor block duration was significantly longer in group 2 than in groups 1 and 3 (P <.05). There were no significant differences in analgesia between groups 1 and 2. Analgesia duration was significantly longer in groups 1 and 2 than in group 3 (P <.05). Heart rate from the 3rd to 6th minute was higher in group 2 than in groups 1 and 3 (P <.05). Systolic arterial pressure from the 3rd to 5th minute and diastolic arterial pressure from 2nd to 6th minute were higher in group 2 than in groups 1 and 3 (P <.05). CONCLUSIONS: Low-dose epinephrine offers more stable hemodynamics and similar blockade, and thus may be beneficial for patients undergoing forearm and hand surgery who are at risk for tachycardia and/or hypertension. PMID- 14556130 TI - Early experience with continuous cervical paravertebral block using a stimulating catheter. AB - BACKGROUND AND OBJECTIVES: This study reports our early experience with continuous cervical paravertebral block (CCPVB) using a stimulating catheter for the management of acute pain after shoulder surgery. METHODS: This prospective observational study presents 256 CCPVB for pain relief after 14 different shoulder operations. Surgery was performed under general anesthesia and blocks were placed prior to induction of general anesthesia (n = 81 [32%]), after induction of general anesthesia (n = 116 [45%]), or postoperatively in the recovery room (n = 59 [23%]). A bolus dose of 30 mL of 0.5% ropivacaine was followed by an infusion of 0.1 mL/kg/h of 0.2% ropivacaine. Patient- or nurse initiated bolus doses of 10 mL of the same drug were used for breakthrough pain and rescue analgesics were available. Postoperative pain, patient satisfaction, and motor function in different parts of the upper limb were evaluated immediately after surgery (time 0), and then 6, 12, 24, 48, 60 hours, and 14 days postoperatively. RESULTS: An average of 2 (range 1-7) attempts were needed to advance the catheter while still stimulating the nerve. Average postoperative pain ranged from 0.27 +/- 1.04 cm to 0.78 +/- 1.56 cm (mean +/- SD) on a visual analog scale (VAS) (0-10 cm) for the first 48 hours and 3.8 +/- 2.1 cm and 3.5 +/ 2.4 cm at 60 hours and 14 days, respectively. Patient satisfaction on a VAS of 0 to 5 was 4.19 +/- 1.1, 4.28 +/- 1.01, and 4.69 +/- 1.05 at times 0, 6 hours, and 14 days, respectively. Motor function returned to normal in the fingers within 24 hours and in the shoulder within 60 hours. Complications included Horner's syndrome (40%), dyspnea (8%), superficial skin infection (5%), posterior neck pain (22%), subclavian artery puncture (1%), contralateral epidural spread (4%), and 8% of the patients complained of an unpleasant "dead feeling" of the arm. Ninety-one percent of patients would request CCPVB again for future shoulder surgery. There was no evidence of nerve damage. PMID- 14556131 TI - No anesthetic or analgesic benefit of neostigmine 1 mg added to intravenous regional anesthesia with lidocaine 0.5% for hand surgery. AB - BACKGROUND AND OBJECTIVES: Neostigmine has shown analgesic benefit when used as an adjunct to epidural or intrathecal anesthesia and analgesia, but evidence of benefit in the peripheral nervous system is controversial. This study aimed to determine if neostigmine 1 mg added to intravenous regional anesthesia (IVRA) provided any advantage in terms of intraoperative anesthesia or postoperative analgesia. METHODS: We recruited 54 patients booked for hand surgery into this randomized, double-blind study. For the IVRA technique, patients were administered 3 mg/kg of 0.5% lidocaine (maximum 45 mL). The treatment group (group N) had 1 mg neostigmine added to lidocaine before dilution. The control group (group C) had no additives to the IVRA solution. At the completion of surgery and after transfer to the recovery room, patients had verbal response pain scores measured at 30 minutes, 1 hour, and 2 hours after cuff deflation. Time to first request for analgesic, side effects, and analgesic consumption at 24 hours were also recorded. RESULTS: Significantly more patients in group N had motor block at 5 and 10 minutes after injection of study solution. There were no other significant differences in sensory block onset, intraoperative anesthesia, postoperative analgesia, or adverse effects between groups. CONCLUSIONS: Neostigmine 1 mg provides no anesthetic or analgesic advantage when added to IVRA for upper limb surgery. PMID- 14556132 TI - Ambulatory perineural infusion: the patients' perspective. AB - BACKGROUND AND OBJECTIVES: Ambulatory perineural local anesthetic infusion is a relatively new method for providing postoperative analgesia, and many aspects of this technique remain in the domain of conjecture and speculation. This retrospective chart review and survey was undertaken to investigate patients' opinions on various aspects of their ambulatory perineural infusion experience. METHODS: Patients who had received an ambulatory perineural infusion from the University of Florida were identified via pharmacy records. Patients were contacted by phone and were asked various questions regarding their experiences and preferences during and after their perineural infusion. RESULTS: Of 217 patients identified, 215 charts were located and retrieved. Of these, 137 (64%) were successfully contacted and 131 (61%) consented to take part in the survey. More than 97% of patients reported that they felt "safe" during home infusion, that one physician telephone call each night was optimal contact, and that they were comfortable removing the catheter with instructions given over the phone. Only 4% would have preferred to return for catheter removal, and 43% felt that they would have been comfortable with only written instructions for catheter removal. CONCLUSION: This investigation suggests that perineural local anesthetic infusion is generally well tolerated by ambulatory patients. PMID- 14556133 TI - Portable infusion pumps used for continuous regional analgesia: delivery rate accuracy and consistency. AB - BACKGROUND AND OBJECTIVES: Multiple benefits of postoperative perineural local anesthetic infusion have been shown including potent analgesia, decreased opioid requirements, and improved rehabilitation. Consequently, portable infusion pumps have been used with increasing frequency to provide perineural infusion for medically unsupervised ambulatory patients. We believe that the infusion rate accuracy and reliability of these pumps infusing potentially toxic doses of medication should be investigated independently. Therefore, we studied the flow rate accuracy and consistency of various portable infusion pumps that have not been examined previously. METHODS: Using a computer/mass balance combination to record infusion rates, 6 pumps (3 electronic and 3 non electronic) were tested. Several factors that may influence pump performance were varied: temperature (ambient/skin), battery (replacement/addition), and catheter exchange (wound/perineural). RESULTS: Infusion rate accuracy differed significantly among the pumps, exhibiting flow rates within +/-15% of their expected rate for 55% to 99% of their infusion duration. Furthermore, the profiles (infusion rate over time) of the various pumps differed significantly depending on the pump power source. Although elastomeric pump infusion rate increased with an increase in temperature, battery life was a limiting factor for one of the electronic pumps. Substituting wound catheters with commonly used perineural catheters did not significantly alter infusion profile. CONCLUSIONS: Factors such as infusion rate accuracy and consistency, infusion profile, temperature sensitivity, and battery life affect the dose of medication administered by various portable pumps used for continuous regional analgesia. Health care providers should take these factors into consideration when choosing and using a portable infusion pump for local anesthetic administration. PMID- 14556134 TI - Altered perceptions after upper and lower extremity blocks: an initial investigation. AB - BACKGROUND AND OBJECTIVES: Nerve blocks frequently produce unusual altered perceptions in the extremities. We examined perceptual changes experienced after peripheral blocks. METHODS: Fifty consecutive patients having an upper or lower extremity block for surgery participated in this prospective study. Patients were divided into 2 groups: upper extremity (n = 20) and lower extremity (n = 30). Each group was asked a list of questions about perceptions of limb sensation, length, weight, and location and given a detailed 2-point discrimination test over the V(1)-V(3) divisions of the trigeminal nerve prior to block and sedation. While the extremity was still blocked, the exam was repeated before postanesthesia care unit discharge. RESULTS: In both groups, 98% of patients described altered limb perception. The perceptions in the upper extremity were: heaviness, 60%; numbness, 50%; warmth, 40%; pain, 30%; full or fat, 20%; floating, 5%; shorter, 0%; or thinner, 10%. The perceptions in the lower extremity were: numbness, 75%; heaviness, 46%; warmth, 33%; pain, 32%; full or fat, 36%; floating, 25%; shorter, 18%, or thinner, 7%. Upper extremity block patients were more likely to describe the limb as lighter (P <.0001); the lower extremity group was more likely to describe the limb as numb (P =.01) or floating (P =.0002). There was no difference in the ability to correctly identify the location of the limb between the groups. There was no difference in 2-point discrimination between each assessment for either group. CONCLUSION: The results of this study confirm and quantify the perceptions experienced by patients undergoing upper and lower extremity blocks. These perceptions are prevalent. This knowledge is helpful in providing patients with accurate preoperative preparation. Further investigation is warranted to determine the neurologic etiology of these observations. PMID- 14556135 TI - Comparison of heart rate response to an epinephrine test dose and painful stimulus in children during sevoflurane anesthesia: heart rate variability and beat-to-beat analysis. AB - BACKGROUND AND OBJECTIVES: During regional anesthesia, various stimuli leading to an adrenergic response can occur. However, simulation of an epidural test dose by using intravenous administration of epinephrine (EPI) has always been compared with an intravenous saline infusion as the control. The aim of this study was to evaluate the possibility of distinguishing in children the effect on HR by an intravascular epinephrine infusion and a painful stimulus, using heart rate variability (HRV) and beat-to-beat analysis of HR. METHODS: Thirty American Society of Anesthesiologists physical status P I children who required elective surgery were studied. At 1 minimum alveolar concentration (MAC) of sevoflurane, electrocardiogram was recorded continuously. Systolic blood pressure (SBP) was measured every minute. Measurements were performed after an intravenous administration of 0.5 microg/kg of epinephrine and during a small skin surgical incision (SI). Time-varying auto-regressive modeling of the interpolated RR sequences was performed for estimating power spectrum (msec(2)). The HF bands were defined by (0.15-0.4 Hz). RESULTS: Median (range) age and weight of all children were 3.5 (1-10) years and 16 (9-30) kg. EPI produced a lower increase in HR than did SI. SBP increased significantly more than did after SI. T-wave amplitude increased significantly after EPI but not after SI. Sixty seconds after the first change in HR, a secondary decrease (in comparison to control value) can be detected with EPI in contrast to SI. HF spectral power increased significantly after EPI administration but decreased after SI. The sensitivity, specificity, and positive and negative predictive value were respectively for DeltaHR >10 beats per minuteof 56%, 26%, 43%, and 38%; for DeltaSBP >15 mm Hg of 60%, 86%, 81%, and 67%; and for DeltaT-wave amplitude >25% of 86%, 73%, 76%, and 84%. Using detection of the secondary decrease of HR, 60 seconds after the first change in HR, sensitivity, specificity, and positive and a negative predictive value were respectively 96%, 100%, 100%, and 96%. CONCLUSIONS: Detection of the secondary HR decrease, 60 seconds after the first change in HR, allows us to distinguish the effects of a painful stimulus from those related to the epinephrine test dose at 1 MAC of sevoflurane. This secondary HR decrease induced by epinephrine appears primarily because of a compensatory increase in parasympathetic tone. PMID- 14556137 TI - Lateral approach to the sciatic nerve block in the popliteal fossa: correlation between evoked motor response and sensory block. AB - BACKGROUND AND OBJECTIVES: The purpose of this study was to identify which of two motor responses of the foot (plantar flexion versus dorsiflexion) best predicts complete sensory blockade of the sciatic nerve when is used for lateral popliteal sciatic nerve block. METHODS: Thirty American Society of Anesthesiologist physical status I or II patients scheduled for foot and ankle surgery under lateral popliteal sciatic nerve block were enrolled in the study. During each block, the needle was placed to evoke one of the following motor responses of the foot: plantar flexion or dorsiflexion. Thirty milliliters of 0.75% ropivacaine was injected after the motor response was elicited at <0.5 mA. The sequence of elicited motor response was randomized. Sensory blockade of the areas of the foot innervated by the deep peroneal, superficial peroneal, posterior tibial, and sural nerves was checked in a blinded manner. Time required for onset of sensory and motor block of the foot was recorded. RESULTS: The 2 groups were similar with regard to demographic variables and type of surgery. The total of nerves blocked (deep and superficial peroneal, posterior tibial, and sural nerves) after elicited plantar flexion was greater (complete sensory block in 58 of 60 nerve distributions) than after elicited dorsiflexion (34 of 60 nerve distributions) (P <.05). Onset of complete sensory and motor blockade of the foot was faster after elicited plantar flexion (16.6 +/- 5.1 minutes, 20.1 +/- 5.1 minutes, respectively) than after elicited dorsiflexion (24.3 +/- 5.1 minutes, 28.1 +/- 5.0 min, P <.05). CONCLUSIONS: After stimulation of the sciatic nerve, plantar flexion better predicts complete sensory blockade of the foot than dorsiflexion when using the lateral approach to the popliteal fossa. The findings of the present study apply to a single injection of 30 mL of ropivacaine 0.75%. PMID- 14556138 TI - The effect of cyclooxygenase-2 inhibitors on bone healing. PMID- 14556136 TI - Computed tomography scanning of the sciatic nerve posterior to the femur: Practical implications for the lateral midfemoral block. AB - BACKGROUND AND OBJECTIVES: Using computed tomography (CT) scans of the thighs, this study addresses sciatic nerve anatomy at the injection site for the lateral midfemoral sciatic nerve block. It addresses the recommendation of neutral leg rotation to facilitate block placement. METHOD: This prospective and descriptive study involves 21 patients scheduled for CT scan imaging of the lower limbs. Transverse CT scans were analyzed at 20, 25, and 30 cm distal to the upper border of the greater trochanter (GT) of the femur with the knee externally rotated by 30 degrees. The angle alpha formed by the broad axis of the sciatic nerve and the coronal plane, skin-to-nerve distance, great vessel-to-nerve distance, division of the sciatic nerve, and widths of the perineural space were assessed. Values are expressed as mean +/- SD. RESULTS: At 20, 25, and 30 cm distal to the GT, the alpha angle was 50 degrees +/- 14 degrees, 55 degrees +/- 13 degrees, and 56 degrees +/- 26 degrees, respectively. This angle increased to nearly 90 degrees when the knee was rotated to a neutral position. The skin-to-nerve distance was 5.9 +/- 1.1 cm, 5.4 +/- 0.9 cm, and 5.7 +/- 1.1 cm. The section width of the perineural space was 1.8 +/- 0.9 cm(2), 3.9 +/- 2 cm(2), and 5.6 +/- 2.4 cm(2). The sciatic nerve was divided in 27% of subjects at 20 cm and in 90% at 30 cm distal. CONCLUSION: The characteristics of sciatic nerve anatomy described in this study support observations and clinical recommendations regarding lateral midfemoral sciatic nerve block. PMID- 14556139 TI - Memories of the early years of regional anesthesia for childbirth. PMID- 14556141 TI - Severe phantom leg pain in an amputee after lumbar plexus block. AB - OBJECTIVES: To describe the onset of phantom leg pain in an amputee with the performance of a lumbar plexus block and the subsequent alleviation after the performance of a sciatic nerve block. CASE REPORT: A 72-year-old American Society of Anesthesiologists physical status III woman presented for left total hip arthroplasty. Her history was significant for a left below the knee amputation. Since the amputation she had suffered from intermittent phantom leg pain. A lumbar plexus block was performed for postoperative pain management. After the lumbar plexus block, the patient experienced severe pain radiating to the left phantom foot. Because of the severity of the phantom pain, a sciatic nerve block was performed. The phantom leg pain resolved within 5 minutes. The intraoperative care under general anesthesia was uneventful. After surgery the patient had continued blockade in both nerve distributions with excellent analgesia. Full recovery of the lumbar plexus and sciatic nerve function was present on the first postoperative day. CONCLUSION: The temporal relationship between the onset of the phantom leg pain and the lumbar plexus block suggests a causal relationship. In this case, it appears that ongoing peripheral input from the lumbar plexus may have been sufficient for the tonic inhibition of phantom pain in the sciatic distribution. The immediate reactivation of the phantom pain and its subsequent relief suggests dynamic processing of peripheral inputs by central neurons, which apparently is rapid and reversible in some cases of phantom pain. PMID- 14556140 TI - Transverse myelitis associated with Acinetobacter baumanii intrathecal pump catheter-related infection. AB - OBJECTIVE: To describe a late neurologic complication of intrathecal pump implantation and show the methods used for the diagnosis and successful treatment of transverse myelitis in this setting. CASE REPORT: A 32-year-old man with a chronic abdominal pain syndrome presented with right lower-extremity numbness 2 months after the placement of an intrathecal morphine pump. This progressed to bilateral lower extremity and ascending sensory loss to T12-L1 dermatome, significant lower-extremity weakness, constipation with overflow incontinence, and detrusor instability causing urinary incontinence in discrete episodes over the following 2 months consistent with a myelopathy. Magnetic resonance imaging (MRI) of the thoracic spine and cerebrospinal fluid (CSF) analysis were consistent with transverse myelitis. The intrathecal pump was removed and an Acinetobacter baumanii catheter-tip infection was diagnosed. Clinical course improved with the co-administration of intravenous corticosteroids and antibiotics, with significant clinical improvement within 30 days. CONCLUSIONS: Clinicians should recognize transverse myelitis as a possible late complication of intrathecal pump placement. Early medical intervention and removal of the intrathecal pump may be necessary to prevent irreversible spinal cord damage and may support good recovery. PMID- 14556142 TI - Ultrasound-guided popliteal block demonstrates an atypical motor response to nerve stimulation in 2 patients with diabetes mellitus. AB - BACKGROUND AND OBJECTIVES: Nerve stimulation is a useful technique to identify peripheral nerves before blockade. We report 2 cases of the failure of nerve stimulation to accurately localize the sciatic nerve in patients with diabetes mellitus undergoing outpatient foot procedures. We also introduce a novel approach to performing a popliteal fossa block using ultrasound guidance. CASE REPORT: Ultrasound-guided popliteal fossa blocks were performed in 2 patients with diabetes mellitus. Both patients failed to develop an appropriate motor response or paresthesia to nerve stimulation. The needle positions were confirmed by ultrasound guidance and injections of local anesthesia were made uneventfully. Appropriate surgical anesthesia was established and the procedures were performed uneventfully. CONCLUSION: Ultrasound facilitated the accurate localization of the sciatic nerve in 2 patients with diabetes mellitus. Neither patient had a paresthesia or muscle twitch below 2.4 mA. There is theoretical concern that patients with underlying neuropathy, such as patients with diabetes mellitus, may have an altered response to either motor or sensory stimulation. PMID- 14556143 TI - Are nonstimulating catheters really inappropriate for continuous nerve block techniques? PMID- 14556145 TI - A case of arteriosclerosis obliterans with monitored regional oxygen saturation during treatment with a lumbar sympathetic ganglion block. PMID- 14556146 TI - Identification of coracoid process for infraclavicular blocks. PMID- 14556147 TI - System failure versus physician accountability. PMID- 14556148 TI - Knotted peripheral nerve catheter. PMID- 14556149 TI - Sacroiliac joint pain: should physicians be blocking lateral branches, medial branches, dorsal rami, or ventral rami? PMID- 14556185 TI - [46,XY female sex reversal patient with a novel point mutation in the coding sequence of the SRY gene]. AB - OBJECTIVE: To investigate the molecular mechanism of a Chinese patient with 46, XY sex reversal. METHODS: DNA fragments of the SRY gene from the typical XY female sex reversal patient and her father were amplified by polymerase chain reaction (PCR). The amplified PCR fragments were cloned into the pUCm-T vector, and direct sequencing was carried out on an ABI 377-3 automated DNA sequencer to detect the mutation. PCR-restriction enzyme digestion was applied to detect the results of DNA sequencing. RESULTS: A novel mutation of the SRY gene was identified in the XY sex reversal patient of this study. A T is replaced by an A in codon 129 at position +387, resulting in the replacement of the polar amino acid tyrosine (TAT) by the stop code (TAA) in the HMG-box, whereas her father was proved to have the wild-type sequence. Because the mutation introduced an enzyme site of MaeIII, the PCR-restrict enzyme digestion showed that there were three bands (131 bp,231 bp and 247 bp) in the patient, whereas two bands (131 bp and 478 bp) in normal man. It verified the results of sequencing analysis. The results after searching the Human Gene Mutation Database showed that this mutation was not described before and should be a new mutation. CONCLUSION: The novel mutation in SRY gene has provided valuable information for the understanding of molecular mechanism of the patient with 46,XY female sex reversal. PMID- 14556186 TI - [Mutation analysis of fibroblast growth factor receptor 3 gene in an achondroplasia family]. AB - OBJECTIVE: To clarify the patients' pathogenic mechanism in an achondroplasia family not according with the genetic law of autosomal dominant inheritance disease at gene level. METHODS: Genomic DNA from peripheral blood of all members in this family was used for amplification of the exon 10 of fibroblast growth factor receptor 3(FGFR3) gene by PCR; mutation was detected by DNA sequencing and identified by restriction endonuclease MaeIII. RESULTS: A new mutation of A to T at nucleotide 1180 was found in patients but not in unaffected members. CONCLUSION: Combined with pedigree analysis, it was summarized that achondroplasia patients in this family might result from this new mutation. PMID- 14556187 TI - [Comparison and analysis of the molecular character of breakpoints in introns of deletion hotspots of dystrophin gene]. AB - OBJECTIVE: To study the exons deletion mechanisms for dystrophin gene, the molecular characters of breakpoints of junction fragments for deletion-type Duchenne muscular dystrophy (DMD) patients with 46 and 51 exons deletion were compared and analyzed. METHODS: Deletion-type DMD patients were detected by multiplex polymerase chain reaction(mPCR). The breakpoints of junction fragments with 46 and 51 exons deletions were cloned and sequenced respectively. RESULTS: Analysis of sequences of deletion-junction fragment of exon 46 showed that the 5'breakpoint was located in AT-rich region of intron 45 and the 3' breakpoint was in medium reiteration repeats (MER1) sequence. There existed 2 bp(ta) junction homology between two breakages. No small insertion, small deletion or point mutation was located near the junction point. Similarly, analysis of sequences of deletion-junction fragment of exon 51 showed that the 5 breakpoint was located in transposon-like human elements (THE1) of intron 50 and the 3' breakpoint was in L2 sequence. There existed 3 bp(cta) junction homology between two breakages. No small insertion, small deletion or point mutation was located near the junction point. By analyzing the secondary structure of junction fragments with 46 and 51 exons deletions, it was demonstrated that all breakpoints of junction fragments were located at the non-matching regions of single-strand hairpin. CONCLUSION: By comparing the junction fragments with 46 or 51 exons deletion, it was found that all of breakpoints were located in repeat sequences and the repeat sequences formed the single-strand hairpin which could make the introns instable and result in exon deletion. PMID- 14556188 TI - [Preclinical research of a new therapy for Gaucher's disease with F213I mutation]. AB - OBJECTIVE: To design and make trial of a new therapy for Gaucher disease. METHODS: A substrate analogue of beta-Glc (glucocerebroside analogue, GCA) was used as a molecular chaperon. Normal and mutant skin fibroblasts were cultured with or without GCA. The activity of beta-Glc was assayed by fluorescent enzymologic techniques. The amount of beta-Glc was determined using Western blot. The beta -Glc was localized by double cell stain experiment. The degradation of glucocerebroside was assessed by thin layer chromatography (TLC) experiment using 14C-Serine. RESULTS: It was found that GCA could enhance the activity and amount of beta-Glc with F213I mutation. It also promoted the beta-Glc with F213I mutation to the lysosome and accelerated the degradation of glucocerebroside. CONCLUSION: The low molecular compound analogous to beta-Glc substrate (GCA ) may be a new therapeutic strategy for Gaucher's disease with F213I mutation. PMID- 14556189 TI - [Studies on molecular epidemiology of Y chromosome azoospermia factor microdeletions in Chinese patients with idiopathic azoospermia or severe oligozoospermia]. AB - OBJECTIVE: To determine microdeletion loci and the characteristic of Y chromosome azoospermia factor(AZF) associated with Chinese idiopathic azoospermia or severe oligozoospermia and hence to provide a theoretic basis for gene diagnosis of AZF microdeletion in Chinese infertility men. METHODS: The subjects of this study included 134 cases of azoospermia and 118 severe oligozoospermia, and 210 healthy male controls. With multiplex PCR technique, an analysis of 15 sequence tag sites(STS) in AZFa, AZFb and AZFc microdeletion was performed. RESULTS: No microdeletion was detected in controls whereas 13 STS loci microdeletions existed in cases, including sY84 and sY86 in AZFa; sY121, sY123, sY124, sY127, sY134 and sy133 in AZFb; sY152, sY242, sY254, sY255 and sY157 in AZFc. Five azoospermia patients were involved in AZFa microdeletions, 7 azoospermia and 3 severe oligozoospermia patients in AZFb, and 14 azoospermia and 18 severe oligozoospermia patients in AZFc. The prevalence rates of microdeletion in AZFa, AZFb and AZFc were 2.0%,4.0% and 12.7% respectively. The microdeletion rate showed statistically significant difference between the cases and controls. CONCLUSION: This study revealed an association between the microdeletion of 13 STS loci of AZF region and the idiopathic azoospermia or severe oligozoospermia in Chinese. There is no evidence to identify the genetic polymorphism of the above STS loci. The frequency and distribution characteristic of AZF microdeletion are similar to those of Caucasians. As candidate loci the 13 STS are useful in clinical gene diagnosis for the detection of AZF microdeletion in Chinese idiopathic azoospermia and severe oligozoospermia. PMID- 14556190 TI - [Development of salivary gland tumors in pleomorphic adenoma gene 1 transgenic mice]. AB - OBJECTIVE: Activation and overexpression of pleomorphic adenoma (PLAG1) gene due to t(3;8)(p21;q12) translocation are associated with the development of human pleomorphic adenomas of the salivary glands. This study was conducted to generate ubiquitously-expressed or tissue-specific expressed PLAG1 transgenic mice and to elucidate the role of PLAG1 gene in tumorigenesis in vivo. METHODS: Human PLAG1 cDNA was cloned from salivary gland tumor or placenta tissues by RT-PCR. Ubiquitous expression vector pCMV-EGFP/PLAG1 driven by CMV promoter and tissue specific expression vector pMMTV-PLAG1 driven by MMTV LTR were constructed. NIH3T3 cells transiently transfected with pCMV-EGFP/PLAG1 showed high expression of PLAG1 in nucleus. Transgenes were microinjected into pronucleus of zygotes to generate transgenic mice. RESULTS: It was found that the human PLAG1 cDNA cloned from several salivary gland tumor and normal placenta tissues consistently showed a variation of a single nucleotide at the same position when compared with the human PLAG1 cDNA sequence in Genbank (Accession No. U65002), which led to T458P at protein level. It might be a single nucleotide polymorphism (SNP)locus. Fused EGFP/PLAG1 protein was found to be localized in the nucleus of NIH3T3 cells transiently transfected with pCMV-EGFP/ PLAG1. Several pCMV-EGFP/PLAG1 and pMMTV PLAG1 transgenic mouse lines were obtained respectively. As might be expected, pMMTV-PLAG1 transgenic mice spontaneously developed salivary gland tumors in three independent lines, among which, line 42 showed tumorigenic phenotype in 100% of transgenic mice within three months after birth. CONCLUSION: Overexpression of PLAG1 gene plays a crucial role in tumorigenesis of salivary gland tumors. PMID- 14556191 TI - [Killer Ig-like receptor gene content diversity and haplotype analysis in Chinese Han population in Shanghai]. AB - OBJECTIVE: To detect the diversity of killer Ig-like receptor(KIR) gene content and the combination of haplotypes in Chinese Han population in Shanghai area. METHODS: DNA samples from 87 randomly unrelated healthy individuals in Shanghai Han population were genotyped with SSP/PCR method. RESULTS: (1) Frequencies of KIR genes: All of 18 known KIRs genes, such as 2DL1-5, 2DS1-5, 3DL1-3, 3DS1, KIR1D and the pseudogenes X, Xv and Z(KIR2DP1) were observed in Shanghai Hans. All individuals contain 3DL3, 2DL4, 3DL2 and 3DL1; the most common genes were 2DL3, Z, 2DL1 and X; the following were 2DS4, 1D, 2DL5, 2DS1, 3DS1 and 2DS5; the next were 2DS2, 2DL2, 2DS3 and Xv. (2) Frequencies of KIR gene haplotypes; there were 13 haplotypes detected in 87 Han individuals, among them, the most frequent one was type 2 (haplotypeA-2DS4). (3) Frequencies of KIR genotypes: 18 kinds of the combinations of the haplotypes were observed; the most frequent ones were AJ(2,2), AF (1,2). Also, In this study were identified five new genotypes FZ1 2 9 , FZ2 1 16 , FZ3 6 17 , FZ4 4 13 and FZ5 2 6 ,which had not been observed in Caucasians so far. CONCLUSION: These findings suggest that there are distinctive frequencies of KIR gene content, haplotype as well as genotype in Chinese Han population in Shanghai area. PMID- 14556192 TI - [Preliminary analysis of mutations in X-linked adrenoleukodystrophy gene(ABCD1) in Chinese patients]. AB - OBJECTIVE: To detect the mutations in exon 6 of ABCD1 gene encoding adrenoleukodystrophy protein(ALDP) in Chinese X-linked adrenoleukodystrophy (ALD MIM 300100) patients. METHODS: Genomic DNA from 14 unrelated patients and two patients' parents with X-linked ALD was extracted using standard procedures from the peripheral blood leukocytes. Polymerase chain reaction (PCR) and DNA direct sequencing were employed to analyze exon 6 of ABCD1 gene. RESULTS: Three mutations in exon 6 were identified in 3 of 14 patients. One mutation was deleted 1 base pair at splice acceptor-site (1489-6 del C). It was not clear what the effect of this mutation is on the ALD protein, maybe induce splicing error. One missense mutation: T1559A(L520Q). These two patients' mothers were heterozygous. The third patient had a mutation: G1548A (L516 L), which is a known polymorphism. It was not a disease causing mutation, so there should be another mutation in this patient. CONCLUSION: For the first time, mutations in ABCD1 are identified in Chinese ALD patients in the mainland of China. No major gene deletion or rearrangement is detected in exon 6. Despite many mutations having been identified in patients with these clinical phenotypes, the genotype-phenotype correlations have not been clarified, suggesting that other genetic or environmental factors may also be involved in determining phenotypic expression in ALD. Two carriers are also confirmed. PMID- 14556193 TI - [The application of denaturing high performance liquid chromatography in detection of novel polymorphisms in PAI-1gene]. AB - OBJECTIVE: To detect single nucleotide polymorphisms(SNPs) in codon region of plasminogen activator inhibitor-1 (PAI-1) gene and associate them with development of coronary artery disease(CAD). METHODS: Denaturing high performance liquid chromatography(DHPLC) and DNA sequencing were used to scan all of exons in PAI-1 gene in 93 CAD patients and 123 control subjects. RESULTS: Two polymorphisms in exon 2, G43A and G49A, which results in missense mutation Ala15Thr and Val17Ile, were found. Heterozygotes for these variants as well as wild type homozygotes were only found in all subjects. The association of genotypes with CAD development and with PAI 1 antigen level were evaluated. The comparison of genotype frequency distribution between two groups revealed that there were more heterozygous carriers of mutation at both polymorphic sites in control than in CAD subjects, but there was no statistically significant difference between them. And genotypes of both polymorphisms were not associated with PAI-1 level. CONCLUSION: Two SNPs in exon 2 of PAI-1 gene were identified by DHPLC method. Both polymorphisms were not associated with the incidence of CAD. PMID- 14556195 TI - [Genetic polymorphism of 8 STR loci on short arm of chromosome 3]. AB - OBJECTIVE: To get the genotype and allele frequency distributions of 8 short tandem repeat (STR) loci on chromosome 3p (D3S1297, D3S1489, D3S1266, D3S1568, D3S1289, D3S1300, D3S1285 and D3S3681) in Chinese Han population in Hunan area. METHODS: Blood samples were collected from the random Han individuals in Hunan and the whole genomic DNA was extracted. STR loci were amplified by multiplex-PCR technique and genotyped by ABI 377 sequencer. RESULTS: Ninety-one alleles were detected, with frequencies ranging from 0.002 to 0.431, and these alleles constituted 312 genotypes. All the 8 loci met Hardy-Weinberg equilibrium. The statistical analysis of 8 STR loci showed the heterozygosity (H) >or= 0.729, the discrimination power (DP) >or= 0.725, the probabilities of paternity exclusion (PPE) >or= 0.596, and the polymorphic information content (PIC >or= 0.682). The result indicated that there was a significant difference between Han ethnic group and the white and the black. CONCLUSION: These results could serve as valuable data to enrich the Chinese genetic database and play an important role in Chinese population genetic and forensic medical application. PMID- 14556194 TI - [Research on expression and control of p16 and p21 by wild-type p53 gene in two lung adenocarcinoma cell lines]. AB - OBJECTIVE: To evaluate the potential of p53 gene therapy for lung cancer in nude mice. METHODS: Two lung adenocarcinoma cell lines L-18 and 95D were infected with adenovirus encoding wild-type p53 gene pAdCMV -p53 (Ad-p53 ) in vitro and in vivo. The antitumor effect of wild type p53 gene was assessed by cell growth curve, reverse transcriptase polymerase chain reaction (RT-PCR) analysis and TUNEL staining methods. RESULTS: The p53-specific growth inhibition and apoptosis of tumor cells were observed in both cell lines in vitro. By RT-PCR analysis, the increasing expression of p21 gene but not of p16 gene after p53 gene infection suggested that p21 gene played an important role in p53 gene induced cell apoptosis. The in vivo study revealed that celiac injection of p53 gene significantly inhibited the tumorigenesis in 95D and L-18 cells in nude mice. However, no obvious inhibition of tumorigenesis was observed after subcutaneous injection of p53 gene in L-18 cell line, compared with the inhibition noted in 95D cell line. CONCLUSION: The results showed the adenovirus-mediated antitumor therapy by means of p53 gene infection might be a potential way to inhibit cancer growth and induce tumor cell apoptosis. PMID- 14556196 TI - [Association of the polymorphism of platelet membrane glycoprotein I a gene with myocardial infarction]. AB - OBJECTIVE: Platelet membrane glycoprotein (GP) Ia/IIa complex is the major collagen receptor on platelets. Platelet activation by GP Ia/ IIa dependent adhesion leads to cellular events that catalyze prothrombin conversion and fibrin clot formation. Correlation between the polymorphism of platelet membrane GP Ia gene and myocardial infarction (MI) was explored. METHODS: A total of 137 patient s with myocardial infarction and 175 controls with no history of coronary heart disease, thrombogenic and hemorrhagenic diseases were studied by case-control. Platelet GP I a gene 807 C/T polymorphisms were checked by polymerase chain reaction-sequence specific primers. RESULTS: There were significant differences in the distribution of T and C alleles between MI and control groups (T:42.70% vs 32.00%, C:57.30% vs 68.00%, P<0.001). No matter among all subjects or among subjects aged 0.05). The same result was seen after stratification by age and gender in each group, respectively(P>0.05). The DD genotype and D allele showed a tendency to decrease with the increase of age (P<0.001). CONCLUSION: The above results suggested that essential hypertension was not associated with ACE I/D polymorphism. The distributions of ACE genotype and allele varied with age, and the subjects with the character of DD genotype were at higher risk of early death. PMID- 14556203 TI - [Analysis on mutations of GJB2 gene in Chinese population]. AB - OBJECTIVE: To determine the prevalence and types of GJB2 mutations and to investigate the genetic mechanism in Chinese autosomal recessive deafness. METHODS: The subjects were four Chinese pedigrees (39 individuals) and 50 normal adults. GJB2 was amplified by PCR. The products were digested with restriction enzyme Apa I, then sequenced. RESULTS: Homozygous deletion C at position 232-235 of GJB2 (235delC),which resulted in frameshift mutation, was found in four affected individuals of two pedigrees; the compound heterozygous deletions (235delC/232G to A) were found in two affected individuals in one pedigree. One carrier with 235delC was found in normal controls (1% allele). Two kinds of polymorphisms 79G to A(V27I) and 3 41A to G(E114G) were found in both affected and normal controls. The frequencies of allele for 79G to A and 341A to G in normal controls were 30%, 21%, respectively. CONCLUSION: 235delC mutation of GJB2 was related with Chinese autosomal recessive deafness, and the 232G to A(Ala78Thr) missense mutation was found to be a novel mutation. PMID- 14556205 TI - [Preimplantation genetic diagnosis for beta-thalassemia using whole genome amplification]. AB - OBJECTIVE: To achieve pregnancy with unaffected embryo using in vitro fertilization and embryo transfer (IVF-ET) and preimplantation genetic diagnosis(PGD) for the couples at risk of having children with beta-thalassemia. METHODS: A couple carrying different thalassemia mutations of codon 41/42 and codon IVS2 position 654 received standard IVF treatment and intracytoplasmic sperm injection, embryo biopsy, single cell polymerase chain reaction and DNA analyses, and only the unaffected or carrier embryos were transferred to uterus. Pregnancy confirmation, and prenatal diagnosis were done at 20 week's gestation. RESULTS: A total of 13 embryos were analyzed in the IVF cycle. PGD indicated that 2 were normal 18.1 , 3 were affected 27.3 , and 6 were carriers 54.5 ; diagnosis was not possible in 2. Three embryos were transferred to uterus on the third day after oocyte retrieval. Ultrasonography showed twin pregnancy with one blighted ovum. The prenatal diagnoses revealed that both fetuses were unaffected, one normal baby and one carrier were born. CONCLUSION: These studies represent the successful application of PGD for beta-thalassemia in China. PMID- 14556204 TI - [Study on gamma-synuclein gene in patients with idiopathic Parkinson's disease]. AB - OBJECTIVE: To evaluate the relationship between idiopathic Parkinson's disease (PD) and two polymorphisms (C243G and A377T) of the gamma-synuclein gene in a Chinese Han population of Shanghai area. METHODS: Polymorphic genotyping was performed with PCR-RPLP technique. Association analysis was carried out in 145 unrelated idiopathic PD patients and 184 age-matched healthy controls. RESULTS: The authors failed to detect any distributional difference of the C243G and A377T polymorphisms of the gamma-synuclein gene between PD cases and control subjects, nor did they find any association. CONCLUSION: These data do not support that gamma-synuclein gene C243G and A377T polymorphisms are involved in idiopathic PD onset in the Han population of Shanghai area. PMID- 14556206 TI - [Technique of PCR-ACRS for the detection of CYP21 gene mutations]. AB - OBJECTIVE: To establish a rapid method of detecting CYP21 gene mutations. METHODS: Fifty Chinese patients with 21-hydroxylase deficiency and some of their families were investigated. Blood samples were obtained for extraction of peripheral blood lymphocytes. A search for restriction sites discriminating between the morbid and the normal in CYP21 gene was made by the computer program DNAssist. PCR-based amplication-created restriction site(PCR-ACRS) was performed at I172N and R356W which are not natural recognition sequence. In addition, I172N and R356W were analysed in five families which conform to the applicability of PCR-ACRS. RESULTS: In 50 identified 21-hydroxylase deficient Chinese patients, 21 were found to have I172 N (3 were homozygote, 18 were heterozygote); 8 were found to have R356W, all of them were heterozygote. By analysing the families, the findings were consistent with the characteristics of autosomal recessive genetic deficiency. CONCLUSION: Analysis of CYP21 gene point mutations using PCR-ACRS is relatively simple, accurate and feasible. PMID- 14556207 TI - [Polymorphism of c-Ha-ras 3'VNTR in hydatidiform mole and its relationship with the development of postmolar tumor]. AB - OBJECTIVE: To study the polymorphism of c-Ha-ras 3' variable number of tandem repeat (VNTR) and its relationship with development of postmolar tumor. METHODS: The cases of hydatidiform mole (HM) were retrospectively analyzed by amplified fragment length polymorphism-polyacrylamide gel electrophoresis. The DNA origin of HM was determined by comparison with the parents' DNA amplified results. RESULTS: Among the samples from 15 cases, DNA from only paternal origin was found in 2 cases. DNA from both parents was in 13 cases, and of these 13 cases, 2 were found to be had balanced DNA origin, 11 had more DNA form paternal origin than DNA from maternal origin. CONCLUSION: The HM which has DNA from both parents origin and predominantly from paternal origin developed to postmolar tumor more frequently. PMID- 14556208 TI - [The SNPs analysis of encoding sequence of interacting factor gene in Chinese population]. AB - OBJECTIVE: To screen the variations of TG interacting factor(TGIF) gene in encoding sequence in Chinese high myopia patients and normal controls and to analyze the SNPs of TGIF gene encoding sequence in Chinese population. METHODS: Genomic DNA was collected from 204 probands with high myopia and 112 unrelated persons without high myopia. The coding sequences of TGIF gene in 316 subjects were analyzed by using exon-by-exon PCR heteroduplex-SSCP analysis and sequencing. RESULTS: There were 3 types of SNP and one single nucleotide mutation in the coding sequence of TGIF gene: IVS-2 nt350 G --> T(36/204), codon140 CCA - > CCG; Pro140Pro codon163 CCG --> CTG;Pro163Leu and codon126 GTG --> GCG; Val126Ala(1/204). The SNPs of codon140 CCA --> CCG and codon163 CCG --> CTG were composed of 3 alleles and 5 genotypes in Chinese population which abide by Hardy Weinberg law. CONCLUSION: There was no evidence to prove that mutations in the TGIF gene are responsible for the high myopia in Chinese. Three SNPs of coding sequence TGIF gene in Chinese population abide by Hardy-Weinberg law. PMID- 14556209 TI - Link between biological signaling and increased enantioseparations of acids using glycopeptide antibiotics. AB - The vancomycin analog A82846B has been shown to provide excellent selectivity as a chiral recognition agent for some acidic test analytes in capillary electrophoresis (CE) and high-performance liquid chromatography (HPLC). In both modes A82846B outperforms vancomycin as a chiral selector. A82846B has enhanced antibacterial activity data in comparison to vancomycin, which is probably due to the increased dimerization constant of over 100 in magnitude in comparison to vancomycin. The link between the electrophoretic and chromatographic separations observed and the biological activity of A82846B is discussed. Dimerization of A82846B in solution was proposed as a theory as to why A82846B gave such enhanced separations for acidic racemates. Further literature and experimental studies support the theory. PMID- 14556210 TI - Absolute configuration determination of chiral molecules in the solution state using vibrational circular dichroism. AB - Advances in the measurement, calculation, and application of vibrational circular dichroism (VCD) for the determination of absolute configuration are described. The purpose of the review is to provide an up-to-date perspective on the capability of VCD to solve problems of absolute stereochemistry for chiral molecules primarily in the solution state. The scope of the article covers the experimental methods needed for the accurate measurement of VCD spectra and the theoretical steps required to systematically deduce absolute configuration. Determination of absolute configuration of a molecule by VCD requires knowledge of its conformation or conformational distribution, and hence VCD analysis necessarily provides solution-state conformation information, in many cases available by no other method, as an additional benefit. Comparisons of the advantages and limitations of VCD relative to other available chiroptical methods of analysis are also presented. PMID- 14556211 TI - Using stereocartography for predicting efficacy of stereoinduction by chiral catalysts. AB - A computational method called stereocartography is used to examine regions around chiral catalysts that are most stereoinducing during Diels-Alder reactions. Geometries and atomic charges of catalysts are first generated quantum mechanically. The transition state of the reaction being catalyzed is then computed quantum mechanically and those enantiomeric transition states are used as probes to determine where around the catalyst stereoinduction is optimal. A description of how to treat catalysts with multiple conformations is given. In this article seven catalysts containing a variety of ligand motifs and metals were evaluated. The hypothesis that the region of maximum stereoinduction must be spatially coincident with the site of chemistry for a catalyst to be efficient is upheld. PMID- 14556212 TI - G-quartet self-assembly under osmotic pressure: remote control by vesicle shrinking rather than stress. AB - Does osmotic pressure stimulate assembly or disassembly of supramolecules in vesicles? Self-assembly was conceivable as intravesicular response to osmotic shrinking upon application of extravesicular overpressure, whereas disassembly was conceivable as a response to bilayer stress in hyperosmotic vesicles. Self assembly of guanosine 5'-monophosphates (GMPs) into G-quartets was selected to investigate the nature of remote control of supramolecular chemistry within vesicles by osmotic pressure. Using circular dichroism spectroscopy to selectively detect G-quartets, we found that extravesicular overpressure stimulates intravesicular self-assembly, whereas underpressure stimulates disassembly. G-quartet self-assembly by osmotic pressure exhibited ion-selective metal-cation templation, as expected. The key conclusions are that supramolecular chemistry within vesicles is governed by vesicle shape rather than vesicle stress and that detection of osmotic pressure by CD spectroscopy is an interesting alternative to the commonly used methods based on fluorescence self-quenching. PMID- 14556213 TI - Photoreaction of a racemate proceeding through radical or ion pairs. The effect of circularly polarized light on the enantiomeric excess of recovered reactant. AB - A simple theory has been developed to explain how circularly polarized light will affect the outcome of a photoreaction of a racemic substrate proceeding through ion pairs or radical pairs. One can calculate the enantiomeric excess and amount of recovered substrate at any stage of the reaction. PMID- 14556214 TI - A novel L-neopentylglycine derivative as auxiliary for copper-catalyzed asymmetric Michael reactions. AB - L-Neopentylglycine diethylamide (4a). was prepared from the new unnatural amino acid L-neopentylglycine (1). The utilization of amide 4a as a chiral auxiliary in the copper(II)-catalyzed asymmetric Michael reaction was investigated in comparison with L-valine diethylamide (4b). Cyclic beta-oxocarboxylates 7 react with 4a and 4b to give the respective enaminoesters 8, which were converted with methyl vinyl ketone (9) in the presence of 10 mol% Cu(OAc)(2). H2O at room temperature in acetone to yield the optically active Michael addition products (R)-10a, b with high selectivity independent of the starting enamine. In the case of the seven-membered beta-oxocarboxylate 7c, however, the valine-derived enamine 8f led to higher enantioselectivity for product 10c. Despite the bulkiness of the neopentyl group, the isopropyl group with an alpha-branch has a better stereoinducing effect. PMID- 14556215 TI - Enantioseparation of chiral alcohols by complex formation and subsequent supercritical fluid extraction. AB - The very first application of supercritical fluid extraction (SFE) on enantioseparation of alcohols is discussed. Resolution of three chiral alcohols (trans-2-chloro-cyclohexanol, trans-2-bromo-cyclohexanol, and trans-2-iodo cyclohexanol) were performed by partial complexation with (-)-O,O'-dibenzoyl (2R,3R)-tartaric acid monohydrate (DBTA). DBTA formed diastereomeric complexes with all S,S-enantiomers stable enough to extract the unreacted alcohols with supercritical carbon dioxide. Resolution efficiency increased with the size of halogen substituents, and by the proper selection of molar ratio, pure (-)-R,R trans-2-iodo-cyclohexanol (ee > 99%, yield: 39%) or (+)-S,S-trans-2-iodo cyclohexanol (ee = 98%, yield: 8%) were prepared in one process step. Achieved resolution efficiency values were much higher in all resolution procedures than in any other known enantioseparation of these racemic compounds. The developed method offers an environmentally friendly, efficient alternative of currently applied resolution processes, also on a preparative scale. PMID- 14556216 TI - Effects from hydrogen bonding and rigidity on selectivity in tartaric acid-based chiral selectors for enantioselective liquid chromatography. AB - Tartaric acid-based selectors 1 ((R,R)-O,O'-Bis(dimethylbenzoyl)-N,N'-diallyl N,N'-dimethyl tartaramide) and 2 ((R,R)-N-allyl-O,O'-bis(dimethylbenzoyl) tartarimide) were synthesized, immobilized on silica, and evaluated as chiral stationary phases in enantioselective chromatography. Comparison with the commercially available column Kromasil-CHI-1, based on selector 3 ((R,R)-O,O' Bis(dimethylbenzoyl)-N,N'-diallyl tartaramide) resulted in the conclusion that amide NH functions are essential to chiral recognition on this sorbent. Furthermore, NH functions contribute significantly to retention of analytes except for alcohols. It was also found that flexibility of the benzoyl moieties is essential to selectivity. PMID- 14556217 TI - Absolute configuration of (-)-2,6-dimethyl-10-(p-tolyl)-2,6(E)-undecadiene from Cistus monspeliensis. AB - (-)-2,6-Dimethyl-10-(p-tolyl)-2,6(E)-undecadiene (1) is a major constituent in the essential oil of Cistus monspeliensis, an aromatic shrub common in Mediterranian countries. 1 was isolated by column chromatography, subjected to ozonolyses, and the absolute configuration was determined by enantioselective gas chromatographic correlation with the ozonolysis product of the sesquiterpene hydrocarbon ar-curcumene with known absolute configuration. PMID- 14556220 TI - Association between atmospheric ozone levels and damage to human nasal mucosa in Florence, Italy. AB - We evaluated the effects of urban air pollutants on human nasal mucosa over an 8 month period on 102 subjects living in Florence, Tuscany, Italy. A group of subjects living in a city with a lower level of pollution (Sassari, Sardinia, Italy) was also analyzed. Nasal mucosa cells were harvested by brushing, a noninvasive procedure. Half of the cells were used for genotoxicity studies using the alkaline comet assay, and half for morphological studies. The levels of DNA damage in the nasal mucosa were considerably higher (+73%) in the subjects living in Florence than in Sassari. High levels of atmospheric ozone in Florence air correlated with DNA damage, and to the prevalence of inflammatory pathologies of the upper respiratory tract, although the ozone concentrations were below the Italian recommended attention level. Furthermore, higher levels of DNA damage were correlated with a dysfunction in the ability to maintain a normal epithelial cell structure. These data suggest an association between ozone air levels and damage in the upper respiratory tract. It remains unclear whether ozone itself or other associated pollutants are responsible for the observed alterations. PMID- 14556221 TI - Structure-activity relationships for the mutagenicity and carcinogenicity of simple and alpha-beta unsaturated aldehydes. AB - Aldehydes are important industrial compounds that are used for the synthesis of chemicals and pharmaceuticals and as solvents, food additives, and disinfectants. Because of their reactivity, aldehydes are able to interact with electron-rich biological macromolecules and adverse health effects have been reported, including general toxicity, allergenic reactions, mutagenicity, and carcinogenicity. The cost, time, and number of animals necessary to adequately screen these chemicals places serious limitations on the number of aldehydes whose health potential can be studied and points to the need of using alternative methods for assessing, at least in a preliminary way, the risks associated with the use of aldehydes. A method of choice is the study of quantitative structure activity relationships (QSARs). In the present work, we present QSAR models for the mutagenicity and carcinogenicity of simple aldehydes and alpha-beta unsaturated aldehydes. The models point to the role of electrophilicity, bulkiness, and hydrophobicity in the genotoxic activity of the aldehydes and lend themselves to the prediction of the activity of other untested chemicals of the same class. PMID- 14556222 TI - Investigating the safety of plants used in South African traditional medicine: testing for genotoxicity in the micronucleus and alkaline comet assays. AB - Previous studies indicate that traditional botanical remedies can be valuable for treating human disease. The potential risk from long-term use of such remedies has not, however, been fully investigated, especially in terms of their potential carcinogenic activity. In the present study, 51 South African plant species were selected on the basis of their use in traditional medicine and crude extracts were sequentially prepared from different dried plant parts using dichloromethane followed by 90% methanol. These extracts were tested for genotoxic activity in human peripheral blood lymphocytes using the micronucleus test, with further testing of select extracts using the alkaline comet assay. Screening results indicated the induction of significant numbers of micronuclei by many of the plant extracts. Several samples also induced DNA damage in human white blood cells using the alkaline comet assay. Although a number of these plants are recognised as toxic by traditional healers, several plants that are used in common remedies were found to be genotoxic and potentially dangerous. Environ. PMID- 14556223 TI - Use of the comet assay for studying environmental genotoxicity: comparisons between visual and image analyses. AB - In order to evaluate the applicability of different measurement parameters employed in the comet assay for analyzing environmental samples, fish hepatoma (RTH-149) cells were exposed to concentrations of the model genotoxic agent hydrogen peroxide (H(2)O(2); 1, 5, and 10 microM) and to five water samples from sites along the Kishon River, the most polluted river in Israel. DNA damage was scored in parallel by visual and computer-image (Viscomet) analyses using 12 different parameters. Each parameter exhibited a different profile of responses. The four visual parameters were highly sensitive to the lowest (1 microM) H(2)O(2) concentration (1.8-7.0-fold of the control). At 10 microM H(2)O(2) exposure, the visual parameter, percentage severe damage, showed the highest (40.3-fold) response while four other parameters, tail area, tail extent moment (Viscomet), mean actual tail length and cumulative tail length (visual analysis), also had substantially elevated responses (8-11-fold). We found that the DNA damage induced by field samples was similar in magnitude to the damage induced by 1 microM H(2)O(2), with only some of the parameters being highly sensitive to the damage. Only about one-half of the parameters could distinguish four significant levels of genotoxicity among the five sampling sites, while the remaining parameters detected only three levels. It is concluded that the choice of parameters for analyzing genotoxicity in ecotoxicological studies should be made in accordance with the characteristics of each parameter. PMID- 14556224 TI - Development and characterization of a stable epithelial cell line from Muta Mouse lung. AB - We have isolated and characterized a stable epithelial cell line from Muta Mouse lung that is a suitable complement to the in vivo assay system. The cells are contact inhibited, forming a flat monolayer, and retain several epithelial/pulmonary characteristics. The genome is stable across more than 50 generations, with a modal chromosome number of 78. Spontaneous rates of micronuclei (19.2 +/- 1.4 per 1,000), sister chromatid exchanges (0.25 +/- 0.004 per chromosome), and chromosome aberrations ( approximately 4%) are lower than, or comparable to, other transgenic cell lines currently used in mutagenicity research. Fluorescence in situ hybridization analyses showed that 80% of cells contain three lambdagt10lacZ loci. Slot-blot analyses indicated that the average cell contains approximately 17 transgene monomers. Spontaneous mutant frequency at the lacZ transgene is stable (39.8 +/- 1.1 x 10(-5)), and the direct-acting mutagens N-ethyl-N-nitrosourea and ICR-191 yielded increases in mutant frequency of 6.3- and 3.2-fold above control, respectively. Benzo[a]pyrene (BaP) exposure increased mutant frequency more than 25-fold above control and did not require an exogenous metabolic activation mixture. Inhibition of Cyp1A1 by 5 microM alpha naphthoflavone eliminated BaP mutagenesis. Activation and mutation induction by the heterocyclic amine 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine required a low concentration (0.05% v/v) of exogenous rat liver S9. High activity of alpha, micro, and pi glutathione-S-transferase isozymes appears to confer resistance to the cytotoxic effects of xenobiotics. The cell line is a suitable complement to the in vivo Muta Mouse assay, and provides an opportunity for routine in vitro mutagenicity testing using an endpoint that is identical to that employed in vivo. PMID- 14556225 TI - Evaluation of mutagenic activity in an extract of pepper tree stem bark (Schinus terebinthifolius Raddi). AB - An extract (decoction) from pepper tree stem bark (Schinus terebinthifolius Raddi) is widely used in Brazil as a topical antiinflammatory agent and to cicatrize wounds. The extract contains catechin, tannins, terpenes, flavonoids, and saponins; of these components, both mutagenic potential and antioxidant properties have been ascribed to flavonoids. The mutagenicity of some flavonoids is believed to be associated with the formation of reactive oxygen species and seems to depend on the number and position of hydroxyl groups. In the present study, we evaluated an extract of S. terebinthifolius in a series of cell-free and bacterial assays in order to determine its genotoxic potential. The extract was negative in a cell-free plasmid DNA test, indicating that it did not directly break DNA. Positive results, however, were obtained in the SOS chromotest, in a forward mutagenesis assay employing CC104 and CC104mutMmutY strains of Escherichia coli, and in the Salmonella reversion assay, using strains TA97, TA98, TA100, and TA102. All the bacterial tests were performed without exogenous metabolic activation due to the topical use of this preparation. The results indicate that pepper tree stem bark extract produces DNA damage and mutation in bacteria, and that oxidative damage may be responsible for the genotoxicity. PMID- 14556226 TI - Methylated trivalent arsenicals as candidate ultimate genotoxic forms of arsenic: induction of chromosomal mutations but not gene mutations. AB - Arsenic is a prevalent human carcinogen whose mutagenicity has not been characterized fully. Exposure to either form of inorganic arsenic, As(III) or As(V), can result in the formation of at least four organic metabolites: monomethylarsonic acid, monomethylarsonous acid (MMA(III)), dimethylarsinic acid, and dimethylarsinous acid (DMA(III)). The methylated trivalent species, as well as some of the other species, have not been evaluated previously for the induction of chromosome aberrations, sister chromatid exchanges (SCE), or toxicity in cultured human peripheral blood lymphocytes; for mutagenicity in L5178Y/Tk(+/-) mouse lymphoma cells or in the Salmonella reversion assay; or for prophage-induction in Escherichia coli. Here we evaluated the arsenicals in these assays and found that MMA(III) and DMA(III) were the most potent clastogens of the six arsenicals in human lymphocytes and the most potent mutagens of the six arsenicals at the Tk(+/-) locus in mouse lymphoma cells. The dimethylated arsenicals were also spindle poisons, suggesting that they may be ultimate forms of arsenic that induce aneuploidy. Although the arsenicals were potent clastogens, none were potent SCE inducers, similar to clastogens that act via reactive oxygen species. None of the six arsenicals were gene mutagens in Salmonella TA98, TA100, or TA104; and neither MMA(III) nor DMA(III) induced prophage. Our results show that both methylated As(V) compounds were less cytotoxic and genotoxic than As(V), whereas both methylated As(III) compounds were more cytotoxic and genotoxic than As(III). Our data support the view that MMA(III) and DMA(III) are candidate ultimate genotoxic forms of arsenic and that they are clastogens and not gene mutagens. We suggest that the clastogenicity of the other arsenicals is due to their metabolism by cells to MMA(III) or DMA(III). PMID- 14556227 TI - Single-cell immunohistochemical mutation load assay (SCIMLA) using human paraffin embedded tissues. AB - It would be advantageous to measure mutation load in situ in order to determine the relationship between a high mutation load and increased risk for cancer or other diseases and to evaluate sources of possible mutagen exposure. Previously, in situ mutation detection assays have been plagued with multiple rounds of amplification and high rates of false-positives and false-negatives. The single cell immunohistochemical mutation load assay (SCIMLA) was developed to measure somatic mutation frequency, pattern, and spectrum in normal tissues with a single round of amplification. The P53 gene was utilized as a mutation reporter because of the unusual property that missense mutations often cause P53 protein to accumulate in the cell, allowing the mutant proteins to be detected by immunohistochemical staining. Alternative reporter genes with stabilized mutant proteins may be envisioned. Single cells that stain positively for P53 protein overabundance (red cells) were microdissected from ethanol-fixed and paraffin embedded tissues. A novel stimulated-PCR (S-PCR) protocol permitted successful amplification of a 1.8-kb segment of the P53 gene (i.e., exons 5-9) in 87% of single mammary cells. Subsequent sequence analysis demonstrated that 35% of the amplified red-stained epithelial cells from normal breast tissue have missense mutations at evolutionarily conserved amino acids. Jackpot mutations, presumably due to clonal expansion, were common. False-positive missense mutations at conserved residues were observed in 3% of the clear cells (i.e., without red stain), presumably due to DNA polymerase error in early PCR cycles. The allele dropout rate was measured at 40% of the amplified cells. SCIMLA is applicable to a variety of tissues, utilizes a single amplification of an endogenous gene, displays mutant cells in situ, and may be adapted to other species. PMID- 14556228 TI - Metabolic polymorphisms and the micronucleus frequency in buccal epithelium of adolescents living in an urban environment. AB - Micronuclei and other biomarkers were evaluated in oral cells from 11- to 16-year old girls living in a foster home in the central area of Mexico City. Variables analyzed for possible association with these biomarkers include smoking habits, body mass index, metabolic polymorphisms for NAT1 and GSTM1 and whether the cells were obtained from the cheek or pharynx. The results indicated that individuals having the NAT1*10 homozygous genotype showed a significant increase in chromatin buds and binucleated cells. When the damage in the cheek was compared with damage in the pharynx, a significant increase in micronuclei and binucleated cells was found for the latter tissue in all the individuals analyzed. PMID- 14556229 TI - Patterns of DNA migration in two-dimensional single-cell gel electrophoresis analyzed by DNA breakage detection-fluorescence in situ hybridization. PMID- 14556230 TI - Binary exposure of A549 cells to benzo[a]pyrene and UVC radiation yields enhanced DNA damage in the comet assay but no enhancement of 8-oxo-deoxyguanosine. PMID- 14556231 TI - Evidence that the metabotropic glutamate receptor 5 antagonist MPEP may act as an inhibitor of the norepinephrine transporter in vitro and in vivo. AB - The mechanisms through which blockade of metabotropic glutamate receptors 5 (mGluR5) results in anxiolytic and antidepressant effects are currently unknown. In the present study, we therefore hypothesized that the anxiolytic- and antidepressant-like profile of the noncompetitive mGluR5 receptor antagonist 2 ethyl-6-(phenylethynyl)-pyridine (MPEP) may be mediated by inhibition of the norepinephrine transporter (NET). Accordingly, we first examined the potency of MPEP to bind to or inhibit uptake at the NET as well as the dopamine and serotonin transporters (DAT and SERT, respectively). We also examined the simultaneous in vivo effects of MPEP and desipramine (DMI) on both NE-like oxidation current in the amygdala (AMY) and cell firing in the locus coeruleus (LC) by means of differential pulse voltammetry (DPV) coupled with electrophysiology. MPEP completely displaced the binding of [3H]-nisoxetine on human NET with a pKi of 6.63 +/- 0.02. In addition, MPEP was able to inhibit [3H] NE uptake in LLCPK cells expressing human NET, with a pIC50 of 5.55 +/- 0.09. In vivo DPV data revealed that both MPEP (30 mg/kg i.p.) and DMI (10 mg/kg i.p.) significantly increased NE-like voltammetric responses levels in the AMY, whereas both compounds also significantly decreased cell firing monitored concomitantly from the second microelectrode in the LC. Collectively, the results of the present study provide potential new mechanisms through which MPEP exerts its anxiolytic and antidepressant effects. PMID- 14556232 TI - Stimulation of postsynaptic alpha1b- and alpha2-adrenergic receptors amplifies dopamine-mediated locomotor activity in both rats and mice. AB - Recent experiments have shown that mice lacking the alpha1b-adrenergic receptor (alpha1b-AR KO) are less responsive to the locomotor hyperactivity induced by psychostimulants, such as D-amphetamine or cocaine, than their wild-type littermates (WT). These findings suggested that psychostimulants induce locomotor hyperactivity not only because they increase dopamine (DA) transmission, but also because they release norepinephrine (NE). To test whether NE release could increase DA-mediated locomotor hyperactivity, rats were treated with GBR 12783 (10 mg/kg), a specific inhibitor of the DA transporter, and NE release was enhanced with dexefaroxan (0.63-10 mg/kg), a potent and specific antagonist at alpha2-adrenergic receptors. Dexefaroxan increased the GBR 12783-mediated locomotor response by almost 8-fold. The role of alpha1b-ARs in this effect was then verified in alpha1b-AR KO mice: whereas dexefaroxan (1 mg/kg) doubled locomotor hyperactivity induced by GBR 12783 (14 mg/kg) in WT mice, it decreased it by 43% in alpha1b-AR KO mice. Finally, to test whether this latter inhibition was related to the occupation of alpha2-adrenergic autoreceptors or of alpha2-ARs not located on noradrenergic neurons, effects of dexefaroxan on locomotor hyperactivity induced by D-amphetamine (0.75 mg/kg) were monitored in rats depleted in ascending noradrenergic neurons. In these animals dexefaroxan inhibited by 25-70% D-amphetamine-induced locomotor hyperactivity. These data indicate not only that the stimulation of alpha1b-ARs increases DA-mediated locomotor response, but also suggest a significant implication of postsynaptic alpha2-ARs. Involvement of these adrenergic receptor mechanisms may be exploited in the therapy of Parkinson's disease. PMID- 14556233 TI - Early embryonic death of glutamate carboxypeptidase II (NAALADase) homozygous mutants. AB - Glutamate carboxypeptidase II (EC 3.4.17.21) catalyzes the hydrolysis (Km = 0.2 microM) of the neuropeptide N-acetylaspartylglutamate to yield N-acetylaspartate and glutamate and also serves as a high-affinity folate hydrolase in the gut, cleaving the polyglutamate chain to permit the absorption of folate. N acetylaspartylglutamate is an agonist at the mGluR3 metabotropic receptor and a source of extracellular glutamate through hydrolysis by glutamate carboxypeptidase II. Given the important role of glutamate in brain development and function, we were interested in the effects of a null mutation of glutamate carboxypeptidase II that would potentiate the effects of N acetylaspartylglutamate. The PGK-Neomycin cassette was inserted to delete exons 9 and 10, which we previously demonstrated encode for the zinc ligand domain essential for enzyme activity. Successful germline transmission was obtained from chimeras derived from embryonic stem cells with the targeted mutation of glutamate carboxypeptidase II. Homozygous null mutants did not survive beyond embryonic day 8. Folate supplementation of the heterozygous mothers did not rescue the homozygous embryos. Mice heterozygous for the null mutation appeared grossly normal and expressed both mutated and wild-type mRNA but the activity of glutamate carboxypeptidase II is comparable to the wild-type mice. The results indicate that the expression of glutamate carboxypeptidase II is upregulated when one allele is inactivated and that its activity is essential for early embryogenesis. PMID- 14556234 TI - Dopamine responsiveness to drugs of abuse: A shell-core investigation in the nucleus accumbens of the mouse. AB - The existence of subterritories within the nucleus accumbens has now been widely supported by histochemical, neurochemical, electrophysiological, as well as morphological and ultrastructural studies and suggest specific afferent and efferent systems involved in different behavioral aspects. Microdialysis studies in the rat have consistently shown that most drugs of abuse increase extracellular dopamine levels preferentially in the shell subregion of the nucleus accumbens. The study of the relative roles of NAc subregions may considerably help our understanding of the neurobiological basis of drug addiction. Accordingly, the aim of the present work was to extend the outcome of rat studies to the mouse species. Five major drugs of abuse were systemically and acutely administered to mice with a microdialysis probe implanted in either the shell or the core. A statistical comparison was performed on data transformed as percentage values of baseline dopamine vs. logarithmic values with baseline dopamine as a covariate. Results show a significant increase in dopamine levels in both the shell and core subregions following cocaine, amphetamine, nicotine, ethanol, and morphine treatments. A difference between shell and core after cocaine, nicotine, and morphine was evident when data were analyzed as percent values of baseline. However, such a shell-core dichotomy became no longer significant when ANOVA was applied on the statistically more appropriate logarithmic transformation of data with baseline as a covariate. The significant baseline differences among groups of mice (dopamine levels in the shell significantly lower compared with dopamine levels in the core) may have compromised, at least in part, the statistical procedure usually applied in microdialysis studies. These findings suggest that a careful evaluation of the data is required when subtle changes in extracellular levels of DA are measured. PMID- 14556235 TI - Direct activation by dopamine of recombinant human 5-HT1A receptors: comparison with human 5-HT2C and 5-HT3 receptors. AB - The effects of dopamine (DA) on the function of human 5-HT1A receptors expressed in Xenopus oocytes and CHO-K1 cells were investigated. In addition, the effect of DA on the activation of three different types of human 5-HT receptors (5-HT1A, 5 HT2C, and 5-HT3) were studied comparatively in Xenopus oocyte expression system. Application of 5-HT or DA in oocytes coexpressing 5-HT1A receptors and G-protein activated inwardly rectifying potassium channels (GIRK1) induced inward currents with respective EC50 values of 4.2 nM and 11.2 microM. Maximal responses induced by DA were 85 +/- 4% of maximal 5-HT currents and DA responses were blocked by the specific 5-HT1A antagonist, WAY-100635 (50 nM). In CHO-K1 cells expressing 5 HT1A receptors, 5-HT and DA inhibited the specific binding of selective antagonist [3H]-8-OH-DPAT with IC50 values of 10.2 nM and 1.4 microM, and both 5 HT and DA inhibited the forskolin-induced accumulation of cAMP. In oocytes expressing 5-HT2C receptors, 5-HT and DA induced inward currents with respective EC50 values of 6.2 nM and 67.7 microM. Magnitudes of maximal DA induced currents were 42 +/- 3% of maximal 5-HT responses and blocked by the 5-HT2 antagonist, piperazine (1 microM). In oocytes expressing 5-HT3 receptors, 5-HT and DA induced fast inward currents with respective EC50 values of 2.1 microM and 266.3 microM. Maximal DA induced currents were 37 +/- 3% of maximal 5-HT responses and blocked the specific 5-HT3 antagonist LY-278584 (0.1 microM). Comparison of the potencies and efficacies of 5-HT and DA indicated that the relative potency of DA increased in the order of 5-HT3 > 5-HT1A > 5-HT2C, and relative efficacy increased in the order of 5-HT1A > 5-HT2C > 5-HT3. These results suggest that although DA activates different subtypes of human 5-HT receptors directly, the potency and efficacy of the binding site varies significantly among different receptors. PMID- 14556236 TI - Synaptic plasticity in rat subthalamic nucleus induced by high-frequency stimulation. AB - The technique of deep brain stimulation (DBS) has become a preferred surgical choice for the treatment of advanced Parkinson's disease. The subthalamic nucleus (STN) is presently the most promising target for such DBS. In this study, whole cell patch-clamp recordings were made from 46 STN neurons in rat brain slices to examine the effect of high-frequency stimulation (HFS) of the STN on glutamatergic synaptic transmission in STN neurons. HFS, consisting of trains of stimuli at a frequency of 100 Hz for 1 min, produced three types of synaptic plasticity in 17 STN neurons. First, HFS of the STN induced short-term potentiation (STP) of evoked postsynaptic current (EPSC) amplitude in four neurons. STP was associated with a reduction in the EPSC paired-pulse ratio, suggesting a presynaptic site of action. Second, HFS of the STN generated long term potentiation (LTP) of EPSC amplitude in eight neurons. Although the EPSC paired-pulse ratio was reduced transiently in the first 2 min following HFS, ratios measured 6-20 min after HFS were unchanged from control. This suggests that LTP is maintained by a postsynaptic mechanism. Third, HFS produced long-term depression (LTD) of EPSC amplitude in five STN neurons. LTD was associated with a significant increase in EPSC paired-pulse ratios, indicating a presynaptic site of action. These results suggest that HFS can produce long-term changes in the efficacy of synaptic transmission in the STN. HFS-induced synaptic plasticity might be one mechanism underlying the effectiveness of DBS in the STN as a treatment of advanced Parkinson's disease. PMID- 14556237 TI - Amygdaloid D1 receptors are not linked to stimulation of adenylate cyclase. AB - In contrast to the classic signal transduction of D1 dopamine receptors in striatum or molecular expression systems, it has been reported that D1 receptor agonists do not stimulate adenylate cyclase in homogenates of microdissected nuclei of the amygdaloid complex. This article examines this phenomenon in detail to determine if lack of cAMP signaling in the amygdaloid complex is an experimental artifact, or an indication of a marked difference from the well studied basal ganglia terminal fields. Thus, whereas dopamine agonists failed to increase cAMP synthesis in the amygdala, forskolin, guanine nucleotides, or Mg2+ were able to stimulate adenylate cyclase activity in the same preparations. Under several different conditions, caudate preparations responded more robustly than amygdaloid preparations, while amygdala homogenates exhibited higher basal production of cAMP. Whereas the beta-adrenergic agonist isoproterenol was able to stimulate cAMP efflux in membranes from both the caudate and amygdala under a variety of tested conditions, neither dopamine nor fenoldopam (D1 agonist) could stimulate adenylate cyclase in the amygdala. Additionally, while manipulation of Ca2+ and calmodulin affected the differential actions of dopamine in the caudate, no change in these parameters restored sensitivity to dopamine in the amygdala. Together, these data challenge the commonly accepted notion that cAMP is a mandatory signaling pathway for D1 receptors. Because it is now proven that G protein-coupled receptors can signal promiscuously, elucidation of the non-cAMP dependent signaling mechanisms resulting from D1 activation is clearly critical in understanding how this important receptor functions in situ. PMID- 14556238 TI - Activation of metabotropic glutamate receptor 5 is associated with effect of amphetamine on brain neurons. AB - The role of metabotropic glutamate receptor 5 (mGluR5) was explored in mechanisms underlying the action of amphetamine (AMPH). The activity of mGluR5 was monitored by measuring the level of [3H]inositol monophosphates in brain neurons, in response to stimulation of 2-choloro-5-hydroxyphenylglycine (CHPG), a selective agonist of mGluR5. Treatment with 1 microM of AMPH for 1 h or 7 days increased the CHPG (1 mM, 30 min)-evoked phosphoinositide turnover by 46% or 92% and 26% or 84% in cultured cortical and hippocampal neurons, respectively, from that of CHPG only treated cells. When AMPH was present during CHPG application post-1 h or 7 day AMPH incubation, the rate of phosphoinositide hydrolysis in cortical neurons became 121% or 142% higher than that treated with CHPG only. The postnatal day (P) 21 (juvenile) and P60 (adult) rats received three intraperitoneal injections of 5 mg/kg of AMPH or saline daily for 6 days. They were challenged on the eighth day with one dosage and sacrificed 3 h later. Reversible 3H-glutamate binding detected increases of 22-89% in the binding levels of cortex and hippocampus of both ages following the AMPH injections. Increases of 13-18% in the levels of mGluR5 mRNA were seen in the juvenile pyramidal neurons of hippocampal CA1-4, granular cells of dentate gyrus, and ventral thalamic nuclei, as shown by in situ hybridization. The AMPH-induced altered activity of mGluR5 is probably associated with changes in the expression of the glutamate receptors, including mGluR5. AMPH may modify the sensitivity of mGluR5 or interact with the receptor itself. PMID- 14556239 TI - Evaluation of a new norepinephrine transporter PET ligand in baboons, both in brain and peripheral organs. AB - Reboxetine is a specific norepinephrine transporter (NET) inhibitor and has been marketed in several countries as a racemic mixture of the (R,R) and (S,S) enantiomers for the treatment of depression. Its methyl analog (methylreboxetine, MRB) has been shown to be more potent than reboxetine itself. We developed a nine step synthetic procedure to prepare the normethyl precursor, which was used to synthesize [11C]O-methylreboxetine ([11C]MRB). We also developed a convenient resolution method using a chiral HPLC column to resolve the racemic precursor to obtain enantiomerically pure individual precursors that lead to the individual enantiomers (R,R)-[11C]MRB and (S,S)-[11C]MRB. Here we report an evaluation of the racemate and individual enantiomers of [11C]MRB as radioligands for PET imaging studies of NET systems in baboons both in brain and in peripheral organs. The relative regional distribution of the radioactivity after injection of [11C]MRB in baboon brain is consistent with the known distribution of NET. For a NET-poor region such as striatum, there were no significant changes in the striatal uptakes with and without the nisoxetine pretreatment. In contrast, a significant blocking effect was observed in NET-rich regions such as thalamus and cerebellum after injection of racemic [11C]MRB, with an even more dramatic effect after injection of (S,S)-[11C]MRB. These results, along with the fact that there was no regional specificity and no blocking effect by nisoxetine for (R,R) [11C]MRB, suggest the enantioselectivity of MRB in vivo, consistent with previous in vitro and in vivo studies in rodents. PET studies of baboon torso revealed a blocking effect by desipramine only in the heart, a NET-rich organ, after injection of (S,S)-[11C]MRB, but not the (R,R)-isomer. These studies demonstrate that the use of (S,S)-[11C]MRB would allow a better understanding of the role that NET plays in living systems. PMID- 14556240 TI - Characterization of fluoxetine plus olanzapine treatment in rats: a behavior, endocrine, and immediate-early gene expression analysis. AB - A large number of individuals afflicted with psychiatric disorders, particularly depression with psychotic features, do not respond to conventional drug therapy. An option for this phenomenon is to augment a standard selective serotonin (5-HT) reuptake inhibitor with an atypical antipsychotic agent. In this regard, fluoxetine and olanzapine have been used concomitantly for treatment-resistant depression and bipolar depression. Although highly efficacious in terms of producing superior improvement of symptoms across a variety of psychological measures, the motor patterns, endocrine profiles, and intracellular signaling pathways affected by drug augmentation have not been determined. Here we show that fluoxetine (10 mg/kg) plus olanzapine (5 mg/kg) given to rats for 7 consecutive days (i.e., subchronic treatment) alters motor activity and diminishes spontaneous behaviors as measured by spatial position and angular path analyses. In addition, the same drug combination pattern sensitizes peak adrenal corticosterone secretion without altering serum glucose levels. We also show that subchronic fluoxetine and olanzapine exposure suppresses the induction of two immediate-early gene transcription factors (e.g., pCREB and FOS) that are associated with long-lasting changes in synaptic efficacy and structural modifications in the prefrontal cortex, piriform cortex, and hippocampus. These results suggest that fluoxetine plus olanzapine can interact in a fashion not predicted by the currently accepted model of fluoxetine monotherapy and provide insight into the synergistic actions of drug augmentation in patients with treatment-resistant depression. PMID- 14556241 TI - A tribute to Bryan D. Hall: Festschrift 2003. PMID- 14556242 TI - The hedgehog signaling network. AB - In the hedgehog signaling network, mutations result in various phenotypes, including, among others, holoprosencephaly, nevoid basal cell carcinoma syndrome, Pallister-Hall syndrome, Greig cephalopolysyndactyly, Rubinstein-Taybi syndrome, isolated basal cell carcinoma, and medulloblastoma. Active Hedgehog ligand is double lipid modified with a C-terminal cholesterol moiety and an N-terminal palmitate. Transport active Hedgehog from the signaling cell to the responding cell occurs through three mechanisms: 1). formation of multimeric Hedgehog which makes it soluble; 2). function of Dispatched in releasing the lipid-anchored protein from the signaling cell; and 3). movement across the plasma membrane of the responding cell by Tout-velu-dependent synthesis of heparan sulfate proteoglycan. In the responding cell, active Hedgehog binds to its receptor Patched, a 12-pass transmembrane protein, which frees Smoothened, an adjacent 7 pass transmembrane protein, for downstream signaling. Patched and Smoothened may shuttle oppositely between the plasma membrane and endocytic vesicles in response to active Hedgehog ligand. In downstream signaling, Cubitus interruptus (Gli proteins in vertebrates), Costal 2, Fused, and Suppressor of Fused form a tetrameric complex. Cubitus interruptus is a bifunctional transcription regulator. In the absence of active Hedgehog ligand, a truncated transcriptional repressor is generated that binds target genes and blocks their transcription. In the presence of active Hedgehog ligand, a full length transcriptional activator binds target genes and upregulates their transcription. Target genes include Wingless (Wnt gene family in vertebrates), Decapentaplegic (Bone Morphogenetic Proteins in vertebrates), and Patched. The upregulation of Patched expression, resulting in Patched protein at the cell membrane, sequesters Hedgehog and limits its spread beyond the cells in which it is produced. Thus, a balance is created by the antagonism of Hedgehog and Patched, whose relative concentrations alternate with respect to each other. Many more factors that are essential for the hedgehog signaling network are also discussed: Megalin, Rab23, Hip, GAS1, PKA, GSK3, CK1, Slimb, SAP18, and CBP. PMID- 14556243 TI - Genetic syndromes among individuals with mental retardation. AB - Individuals with mental retardation more commonly have malformations and other structural anomalies than individuals without mental retardation. In many cases, the associated anomalies comprise recognizable syndromes caused by genetic or environmental insults. To co-occurrence of structural anomalies with mental retardation thus assists in the diagnostic evaluation, particularly in infants and young children. The coexistence of structural and mental abnormalities also suggests that both originated in the embryonic period of development. PMID- 14556244 TI - X-linked cubitus valgus with mental retardation and typical face. AB - In 1973, Jones and Smith described two maternal male first cousins with a similar pattern of malformation, including mental retardation, cubitus valgus, and unusual facies. The purpose of this report is to describe three additional cases, a 10-year-old male and his 30-year-old maternal uncle and an unrelated 15-year old boy, bringing to five the total number of individuals with this disorder. The principal features include moderate mental retardation, mild microcephaly, a short philtrum, deep-set, downslanting palpebral fissures, multiple nevi, and striking cubitus valgus. Documentation of this disorder in two maternal male first cousins as well as in a male and his maternal uncle support an X-linked recessive mode of inheritance for this condition. PMID- 14556245 TI - A new X-linked syndrome with agenesis of the corpus callosum, mental retardation, coloboma, micrognathia, and a mutation in the Alpha 4 gene at Xq13. AB - We describe two brothers with a unique pattern of malformations that includes coloboma (iris, optic nerve), high forehead, severe retrognathia, mental retardation, and agenesis of the corpus callosum (ACC). Both boys have low-set cupped ears with sensorineural hearing loss, normal phallus, pectus excavatum, scoliosis, and short stature. One brother had choanal atresia and cardiac defects consisting of ventricular septal defect (VSD) and patent ductus arteriosus (PDA) which resolved spontaneously. Differential diagnosis between a number of clinical entities was considered, however, because ACC and the distinctive facial features were reminiscent of FG syndrome, DNA was analyzed for markers linked to the FGS1 locus at Xq13-q21. Notably, the brothers were concordant for markers spanning this presumed FG region, and in both we have identified adjacent alterations ( 57delT and T-55A) in the Alpha 4 gene located within this interval. Alpha 4 is a regulatory subunit of the major cellular phosphatase, PP2A, that has recently been shown to interact with MID1, the product of the gene mutated in X-linked Opitz GBBB syndrome. The double nucleotide change identified in this family was not observed in 410 control chromosomes, suggesting that it may be a pathogenetic change. Altered expression of Alpha 4, through either a change in translational efficiency, mRNA stability or splicing, could explain the clinical phenotype in these boys and the phenotypic overlap with Opitz GBBB syndrome. PMID- 14556246 TI - GTF2I hemizygosity implicated in mental retardation in Williams syndrome: genotype-phenotype analysis of five families with deletions in the Williams syndrome region. AB - Most individuals with Williams syndrome (WS) have a 1.6 Mb deletion in chromosome 7q11.23 that encompasses the elastin (ELN) gene, while most families with autosomal dominant supravalvar aortic stenosis (SVAS) have point mutations in ELN. The overlap of the clinical phenotypes of the two conditions (cardiovascular disease and connective tissue abnormalities such as hernias) is due to the effect of haploinsufficiency of ELN. SVAS families often have affected individuals with some WS facial features, most commonly in infancy, suggesting that ELN plays a role in WS facial gestalt as well. To find other genes contributing to the WS phenotype, we studied five families with SVAS who have small deletions in the WS region. None of the families had mental retardation, but affected family members had the Williams Syndrome Cognitive Profile (WSCP). All families shared a deletion of LIMK1, which encodes a protein strongly expressed in the brain, supporting the hypothesis that LIMK1 hemizygosity contributes to impairment in visuospatial constructive cognition. While the deletions from the families nearly spanned the WS region, none had a deletion of FKBP6 or GTF2I, suggesting that the mental retardation seen in WS is associated with deletion of either the centromeric and/or telomeric portions of the region. Comparison of these five families with reports of other individuals with partial deletions of the WS region most strongly implicates GTF2I in the mental retardation of WS. PMID- 14556247 TI - Is Jaffe-Campanacci syndrome just a manifestation of neurofibromatosis type 1? AB - This article describes four patients with non-ossifying fibromas (NOFs) and multiple cafe-au-lait spots. Two of the patients were diagnosed with NOFs when they presented with a femur fracture. The other two patients were diagnosed with NOFs because of complaints of leg problems. In addition, axillary freckles and Lisch nodules were present in all four patients and multiple cutaneous neurofibromas in two patients. These four patients fulfilled the diagnostic criteria for neurofibromatosis type 1 (NF1) and also have been diagnosed with Jaffe-Campanacci syndrome. We propose that Jaffe-Campanacci syndrome is a manifestation of NF1 and suggest that patients with NF1 should have more rigorous radiographic screening of the long bones during early adolescence or adulthood to determine the presence or absence of NOFs. Appropriate intervention (exercise restriction, bracing, and/or surgery) might decrease the long-term disability associated with Jaffe-Campanacci syndrome. PMID- 14556248 TI - Does chromosome 22 have anything to do with sex determination: further studies on a 46,XX,22q11.2 del male. AB - Several years ago, we presented a patient with true hermaphroditism and partial duplication of chromosome 22 and no evidence of SRY (Aleck et al. [1999: Am J Med Genet 85:2-4]). Recently a 46,XX male with velocardiofacial syndrome and a deletion of 22q11.2 and no evidence of Y chromosomal loci in blood DNA was reported (Phelan et al. [2003: Am J Med Genet 116A:77-79]). We have restudied this patient as he enters puberty. Because chromosomal deletions sometimes involve micro rearrangements of nearby material, we have extensively studied this individual's chromosome 22 looking for evidence of any gene duplication. We studied a number of variable number tandem repeat (VNTR) loci along chromosome 22 in the patient and both parents. Normal Mendelian inheritance of the VNTRs was found. We then used quantitative multiplex PCR of short fluorescent fragments (QMPSF) to delineate the 22q11.2 deletion in this patient (Jacquet et al. [2002: Hum Molec Genet 11:2243-2249]) and found a pattern of deletion typical of the velocardiofacial DiGeorge syndrome. Finally, the patient's DNA has been analyzed using a full coverage human chromosome 22 genomic microarray (array comparative genomic hybridization [CGH]) for evidence of rearrangements outside the classical velocardiofacial DiGeorge associated deletion (Buckley et al. [2002: Hum Molec Genet 11:3221-3229]). The array-CGH profile of this patient confirms the deletion encompassing the typically deleted region associated with the velocardiofacial DiGeorge syndrome and provides no support for additional gene copy number aberrations on 22q. Thus, there is no evidence of any chromosome 22 trisomic material. In this case, the rare events of sex reversal and 22q11.2 deletion may have occurred together by chance. PMID- 14556249 TI - Adult height in Noonan syndrome. AB - Short stature is a very common manifestation of Noonan syndrome (NS) and is accompanied by a variable delay in bone age. Although reports of adult height in NS are uncommon, some feel growth hormone therapy will increase adult height. We report our findings in 73 adults over 21 years of age with NS. Thirty percent of this group had an adult height in the normal range between 10th percentile and 90th percentile. Over half of the females and nearly 40% of males had an adult height below the 3rd percentile. The presence or severity of heart disease was not a factor, and none of the adults with a normal height had been treated with growth hormone. Serial measurements of height for many years through childhood to adulthood were available in only a few patients, but their pattern of growth suggests catch up may occur in late adolescence. To evaluate the benefit of growth hormone therapy, long term serial height measurements over a period of years comparing treated and untreated patients are needed. It will be important to determine what role, if any, the mutated PTPN 11 gene plays in the short stature common in NS. PMID- 14556250 TI - Methotrexate/misoprostol embryopathy: report of four cases resulting from failed medical abortion. AB - Methotrexate, a methyl derivative of aminopterin, is a folic acid antagonist and a known human teratogen; misoprostol is a synthetic prostaglandin E1 analog that causes uterine contractions. Recently, there has been resurgence in the use of methotrexate in combination with misoprostol or of methotrexate alone for the treatment of unwanted or ectopic pregnancies, respectively. This report documents the findings in four infants who were exposed prenatally to methotrexate alone or in combination with misoprostol in a failed attempt at medical abortion or treatment of ectopic pregnancy. All patients demonstrated growth deficiency, with growth parameters <10th centile, and all displayed features consistent with methotrexate and/or misoprostol embryopathy. Since an increasing number of medical abortions are being performed, it is important for physicians to recognize the associated teratogenic effects of these abortifacients. Data from the patients herein described should prompt obstetricians and other health care practitioners who prescribe these medications to counsel their patients regarding these risks, especially if the treatment regimen fails to induce an abortion. PMID- 14556251 TI - Reciprocal translocation associated with multiple exostoses in seven members of a three generation family and discovered through an infertile male. AB - We report a four generations family with multiple exostoses segregating with a reciprocal translocation t(8;19)(q24.11;q13.13) in 8 members of three generations. FISH investigations detected a breakage of the dosage-sensitive EXT1 gene. Although three members of the family died perinatally from unknown causes and one carrier had four spontaneous abortions, the translocation was discovered only when the cytogenetic analysis was requested in an affected male because of oligozoospermia. In fact, it is well known that infertile males may be carriers of reciprocal or Robertsonian translocations with a higher frequency than the general population. This family stresses the importance of requesting the cytogenetic analysis in all cases in which a dominant disease segregates with repeated miscarriages and/or newborn deaths of unknown cause. PMID- 14556252 TI - Toriello-Carey syndrome: delineation and review. AB - Toriello and Carey [1988: Am J Med Genet 31:17-23] first described a syndrome with component manifestations of corpus callosum agenesis, unusual facial appearance, Robin sequence, and other anomalies. This was termed the Toriello Carey syndrome by Lacombe et al. [1992: Am J Med Genet 42:374-376]. Since then, 11 reports describing 16 additional children have been published; in addition, we have had the opportunity to review over 30 unpublished cases. However, for various reasons, only 25 of the unpublished patients were included in this review. Based on this total, we can begin to better delineate this syndrome, as well as provide some information on natural history. PMID- 14556253 TI - Craniofacial-deafness-hand syndrome revisited. AB - In 1983 Sommer described a new syndrome in a mother and her infant daughter which was subsequently called the syndrome of craniofacial, hand anomalies and sensorineural deafness. The syndrome consisted of a normal calvarium with a flat facial profile, hypertelorism and small palpebral fissures with an antimongoloid slant, a depressed nasal bridge with a button tip and slitlike nares and a small "pursed" mouth. Profound sensorineural hearing loss and ulnar deviation of the hands with flexion contractures of digits three, four and five was evident. The family had another child, a son, two years after the birth of the index case that had the exact manifestations as his mother and sister. Because of three affected family members in two generations and a phenotype of midfacial anomalies and dystopia canthorum resembling Waardenburg syndrome, a search for mutations in the PAX3 gene was undertaken. A missense mutation in the paired domain of PAX3 (Asn47Lys) was detected. We have provided a 20-year follow-up of a syndrome characterized by craniofacial anomalies, hearing loss and hand deformities and which is caused by a PAX3 missense mutation. PMID- 14556254 TI - A new recessive syndrome with VATER-like defects, pulmonary hypertension, abnormal ears, blue sclera, laryngeal webs, and persistent growth deficiency. AB - VATER association is a term frequently used to describe children with multiple malformations. However, occasionally these children have malformations that are not typical of VATER defects and may represent unique multiple malformation syndromes with specific prognoses and recurrence risks. Two siblings with a heretofore previously undescribed multiple malformation syndrome are presented whose features include vertebral defects, cardiac abnormalities, pulmonary hypertension, laryngeal webs, blue sclerae, and persistent growth deficiency. Autosomal recessive inheritance is suggested. These cases suggest that other individuals labeled VATER association with multiple other defects should be reviewed more closely to give accurate prognosis and recurrence risk information to families. PMID- 14556255 TI - Normal cognition and behavior in a Smith-Lemli-Opitz syndrome patient who presented with Hirschsprung disease. AB - Smith-Lemli-Opitz syndrome (SLOS) is an autosomal recessive disorder of cholesterol biosynthesis. It is caused by mutations in the gene encoding the enzyme 7-dehydrocholesterol Delta7-reductase (DHCR7), which catalyzes the final step in cholesterol biosynthesis, usually resulting in cholesterol deficiency. We report a 3.5-year-old girl who has cognition in the low average range and normal behavior, but in whom molecular studies identified two missense mutations in DHCR7: V326L and F284L. She was born at term following an uncomplicated pregnancy and delivery, and presented at 12 days of age with poor feeding, abdominal distention, and jaundice. Colonic biopsy was consistent with Hirschsprung disease. On physical examination she had mild ptosis, a long philtrum, mild micrognathia, a short, upturned nose, and subtle 2,3 syndactyly. Her 7 dehydrocholesterol (7-DHC) level was markedly elevated at 8.7 mg/dl (normal 0.10 +/- 0.05), and her cholesterol level was normal at 61 mg/dl (normal for newborn period 50-80 mg/dl). Karyotype analysis was normal, 46,XX. Breast milk feeding was initiated and continued for 18 months. Cholesterol supplementation was implemented at 100 mg/kg/day at 3 months, which resulted in increased cholesterol levels and reduced dehydrocholesterol levels. Neuropsychological testing has shown functioning in the low average range, between the 14th and 18th centiles when compared to peers. This is markedly higher than most children with SLOS. She has no behavioral problems. MRI and MRS testing of the brain revealed no structural abnormalities. This is in contrast to a recently reported case by Prasad et al. [2002: Am J Med Genet 108:64-68] with a mild phenotype, behavioral problems, and abnormal MRI, who is compound heterozygote for both a null and missense mutation. Our case suggests that patients with severe feeding disorders with or without Hirschprung disease and postnatal onset microcephaly may warrant screening for SLOS. PMID- 14556256 TI - Moebius sequence and hypogonadotrophic hypogonadism. AB - A distinct form of Moebius sequence is associated with hypogonadotrophic hypogonadism. There have been five such cases to date. We now add a further case with detailed neurologic, endocrine, and autopsy findings and offer a hypothesis drawing parallels with the already established basis of hypogonadotrophic hypogonadism in the X-linked form of Kallman syndrome. PMID- 14556257 TI - Modifications and substitutions of the RNA extraction module in the ViroSeq HIV-1 genotyping system version 2: effects on sensitivity and complexity of the assay. AB - Genotypic testing for HIV-1 resistance to anti-retroviral drugs has become accepted widely as a routine method to guide anti-retroviral therapy. However, implementation into routine high-throughput laboratory diagnosis is difficult due to the complexity of the assay. A commercially available assay is the ViroSeq HIV 1 Genotyping System (Applied Biosystems, Weiterstadt, Germany). We modified and substituted the RNA extraction module to optimize the proportion of samples amplified successfully as follows: 1 ml plasma was concentrated by ultracentrifugation and extracted according to the manufacturer's instructions (Kit), by substituting the lysis buffer (Roche, Roche Diagnostics GmbH, Mannheim, Germany), and by using the QIAamp Viral RNA Kit (Qiagen GmbH, Hilden, Germany) with elution volumes of 60 (Q60) or 50 micro l (Q50). Overall Q50 showed a higher success rate (97%) than the other extraction modules used (range 88-91%). In samples with a viral load range of 1,000-4,999 copies/ml, Q50 was superior (95 vs. 65% to 83%), while in samples with a viral load range of 5,000-9,999 copies/ml or those with 10,000 or more copies/ml, the success rate of the extraction procedures showed no significant differences. In 18 samples, which were negative using the Kit or Roche extraction, Q60 resulted in 7/18 positive results; in addition the Q50 was successful in amplifying 7/10 of the Q60 negative samples. When investigating samples with a measurable viral load of less than 1,000 copies/ml or lower, Q50 had the highest success rate with 80% compared to the other procedures (33-63%). A statistically significant new cut-off could be defined for Q50 at a value of 250 copies/ml. The results showed clearly that the ViroSeq System is suitable for analyzing the HIV-1 genotype over a wide range of viral loads but could be improved significantly when substituting the RNA extraction module with Q50 without using a nested PCR protocol. This is of great importance as it avoids further time- and cost-intensive steps. PMID- 14556258 TI - Elevated levels of antibodies against 70 kDa heat shock proteins in the sera of patients with HIV infection. AB - Heat shock proteins (Hsp), especially 70 kDa heat shock protein (Hsp70) play an important role in the life cycle of HIV-1 virus. Hsp70 is overexpressed in HIV infected cells and this is the most abundant Hsp associated with HIV virions. The aim of our study was to investigate whether HIV infection increases the extent of specific humoral immune response against Hsp70. The serum concentration of anti Hsp70 IgG antibodies was measured in 47 HIV-infected patients, and 62 healthy, HIV-seronegative persons. Nineteen patients on highly active anti-retroviral therapy (HAART) were followed for 24 months in a longitudinal study. Anti-Hsp70 antibodies were measured by ELISA, using recombinant human Hsp70. Levels of anti Hsp70 antibodies were significantly (P < 0.0001) higher in the HIV-infected patients (median: 1409 (25th-75th percentile: 1031-2214) AU/ml) than in healthy control subjects (626 (429-970) AU/ml). In 19 HIV patients, serum levels of anti Hsp70 antibodies significantly (P < 0.001) decreased during 24 (11-41) months HAART (1309 (887-2213) AU/ml before and 640 (386-959) AU/ml during HAART), accompanied by viral load reduction and CD4+ count elevation. It is concluded that HIV-infection induces a marked increase in the anti-Hsp70 antibody levels, which is consistent with the enhanced expression of Hsp70 on the surface of HIV infected cells and/or incorporation of the protein into the membrane of HIV virions. PMID- 14556259 TI - Effector HIV-specific cytotoxic T-lymphocyte activity in long-term nonprogressors: associations with viral replication and progression. AB - Ex vivo effector cytotoxic T-lymphocyte (CTL) activity was assessed in 27 members of the Australian Long-Term Nonprogressor cohort and correlated with genetic, virological, and immunological markers. The 27 individuals were antiretroviral naive with CD4(+) T-cell counts of >500 cells/ microl for more than 8 years after human immunodeficiency virus type 1 (HIV-1) infection. Effector CTL activity was determined using a standard ex vivo chromium release assay. Individuals with CTL activity (HIV-1 env(IIIB) or pol or gag) were then compared to those without CTL activity in relation to plasma HIV-1 RNA, ICD p24 antigen, beta(2)-microglobulin, CD4 and CD8 T-cell counts, CCR5 and CCR2b genotypes, and progression to CD4 <500 cells/microl or commencement of antiretroviral treatment. Of the 27 individuals examined, 19 had no detectable effector CTL activity. The eight individuals with detectable CTL activity had significantly higher plasma levels of HIV-1 RNA (P = 0.014), immune complex dissociated p24 antigen (P = 0.006), and beta(2) microglobulin (P = 0.009). There was increased risk of progression within 4 years of study entry in individuals with detectable effector CTL activity, higher plasma levels of HIV-1 RNA, higher beta(2)-microglobulin levels, and higher immune complex dissociated p24 antigen levels at enrollment (P = 0.017, P = 0.004, P = 0.027, P = 0.008 respectively). Multivariate analysis demonstrated viral load remained the strongest predictor of disease progression within this group (P = 0.017). There were no significant associations between CTL response and chemokine receptor genotype. These findings demonstrate the importance of HIV replication in generating an effector CTL response and show that effector CTL activity may be an early predictor of progression in people with long-term asymptomatic HIV infection. PMID- 14556260 TI - HBV DNA integration and HBV-transcript expression in non-B, non-C hepatocellular carcinoma in Japan. AB - Few studies have examined the etiology of hepatocellular carcinoma (HCC) in patients without hepatitis virus infection. We evaluated the role of occult hepatitis B virus (HBV) infection in the development of HCC in Japanese patients without hepatitis B surface antigen (HBsAg) and antibodies to hepatitis C antigen (anti-HCV). Twenty-one HBsAg negative and anti-HCV negative (non-B, non-C) patients with HCC were studied. HBV DNA in serum and HBV transcripts in liver were examined by polymerase chain reaction (PCR) or reverse transcription and PCR. HBV DNA integration was examined by Southern blot analysis or cassette ligation-mediated PCR as described previously. p53 mutations were examined by direct sequencing. HBV DNA was not detected in serum from any patients. HBV related transcripts were detected in 5 of 7 HCCs from patients with antibodies to hepatitis core antigen (anti-HBc) and in 3 of 14 HCCs from patients without anti HBc (P = 0.0261). HBV DNA was integrated into human genome in two non-B, non-C HCCs. Of the 14 patients without anti-HBc, 5 had a history of excessive alcohol intake. In exons 5 through 8 of the p53 gene, mutations were detected in 2 of 8 HCCs with HBV-transcripts and in 5 of 13 HCCs without such transcripts. p53 mutation at codon 159 was found in 2 of 6 patients with excessive alcohol intake without HBV-transcripts. These results suggested that occult HBV infection might play an important role in hepatocarcinogenesis in non-B, non-C patients with anti HBc and that excessive alcohol intake might be related to HCC in non-B, non-C patients in Japan. PMID- 14556261 TI - Complete genome sequence and phylogenetic analysis of hepatitis B virus (HBV) isolates from patients with chronic HBV infection in Korea. AB - Although there is a report of high rate of hepatitis B virus (HBV) infection in South Korea, only a few entire genome sequences of HBV isolates from Korea have been reported. To obtain the complete nucleotide sequence of the Korean HBV, viral DNA was extracted from sera of Korean patients with chronic HBV infection who have not been exposed to any antiviral treatment. Complete genomic sequences were determined on three Korean HBV isolates. The entire genomic length of Korean HBV isolates, designated as KUHB84, KUHB81, and KUHB95, was 3,215 base pairs. No deletions and insertions were observed. Core promoter mutations (T1762/A1764) were detected in two isolates, KUHB84 and KUHB95. Phylogenetic analysis based on the entire genomic sequences showed that the Korean HBV isolates were genotype C and related closely to the Japanese HBV. PMID- 14556262 TI - Virological and biochemical relapse according to YMDD motif mutant type during long-term lamivudine monotherapy. AB - Whether the type of lamivudine-resistant virus in hepatitis B virus (HBV) influences the clinical outcome, it is not completely understood. We evaluated the serial changes in YMDD motif mutant in 60 Japanese genotype C-HBV patients who received long-term lamivudine monotherapy. YIDD or YVDD alone tended to stop shifting to the mixed type (YVDD and YIDD) within 12 months after the detection of mutant virus. Hence, the characteristics, virological relapse (DNA breakthrough) and biochemical relapse (breakthrough hepatitis) of 49 patients, who could be classified into three types (continuous YVDD, continuous YIDD, and the mixed type), were investigated. YVDD and YIDD type tended to have the opposite background with regard to age, histology, and viral load. The mixed and YIDD types tended to have similar backgrounds, except for viral load. In the mixed type, both the HBeAg-positive rate and viral load as risk factors for emergence of the mutant tended to be high. Mutant virus, DNA breakthrough and breakthrough hepatitis emerged significantly earlier in the mixed type than the two other types. The incidence of severe breakthrough hepatitis accompanied by icteric flare-up tended to be higher in the mixed type than the other types. Our results suggest that the YMDD motif mutant type might emerge from different backgrounds and modulate the virological and biochemical relapse after the emergence. Large-scale studies of each mutant type should be conducted in the future to confirm these findings. PMID- 14556263 TI - Broadly cross-reactive mimotope of hypervariable region 1 of hepatitis C virus derived from DNA shuffling and screened by phage display library. AB - The hypervariable region 1 (HVR1) is the target of neutralizing antibodies but with isolate specificity. The aim of this study was to obtain immunogenic mimotopes of HVR1, which can react broadly with different HVR1 antibodies and could be one of the candidate immunogens in an effective vaccine against HCV. Thirty-one HVR1 cDNA fragments were digested by DNase I into a pool of random fragments and reassembled by repeated cycles of annealing in the presence of DNA polymerase to their original size. The shuffled HVR1 was then inserted into the gene III phagemid vector pCANTAB-5E and displayed on the surface of the phage. Eight individual phages were selected after four rounds of biopanning against anti-HVR1. ELISA was carried out on immobilized purified phages, respectively, to detect their reactivity with a panel of sera. DNA sequences of the inserts were analyzed and compared with the consensus sequences defined by Puntoriero et al. [(1998) EMBO J 17:3521-3533]. The reactivity of the eight selected clones to the 30 sera were from 53.3 to 80%. Among these, phage 13 (ETYVSGGSAARNAYGLTSLFTVGPAQK, aa 384-410) reacted most broadly. None of the selected sequences encoded for peptides corresponded to known HVR1 from original viral isolates. The two high reactive phages had the similar amino acid sequences with the consensus, which might play a particular role in determining the frequency of reactivity. In conclusion, this study has used effectively DNA shuffling combined with phage display technology to identify broadly cross reactive mimotopes recognized by human polyclonal antibodies. Mimotope 13 and 23 appeared to be most reactive immunologically and could be candidate immunogens. Efforts are now underway to identify their neutralizing antibodies by immunization of animals. PMID- 14556264 TI - Evaluation of antibody-based and nucleic acid-based assays for diagnosis of hepatitis E virus infection in a rhesus monkey model. AB - We have evaluated four hepatitis E virus (HEV) specific antibody assays, using sequential samples taken from 86 rhesus monkeys at intervals for up to 86 weeks after they had been infected with different doses of HEV. The animals are a common experimental model of hepatitis E. The large collection of sequential samples used avoids uncertainties encountered in previous studies regarding the precise infection status of study subjects and minimizes bias due to the individuality of response to infection. One assay (YES IgG) was produced with synthetic peptides; the others (E2 IgM, E2 IgG, and GL IgG) were produced with recombinant antigens. The results were compared with the viral RNA contents of the serum and stool samples and the occurrence of these virological and immunological markers in the course of the infection was temporally related to the development of hepatitis. Diagnostic utility of the markers was assessed according to their response rates and prevalence at different times in the course of infection. All the animals produced E2 IgG and developed viremia and all but one also produced E2 IgM and excreted the virus in stool, whereas response rates for the other antibodies were lower and decreased with virus dose. Hepatitis occurred over a period of 4 weeks between 3 and 7 weeks after infection. Virological activity occurred mainly during the incubation period and the prevalence of viral markers declined rapidly after the onset of hepatitis. Production of the E2 antibodies immediately preceded the onset of hepatitis, and this was followed about one week later by production of the other antibodies. Seroprevalence E2 IgM reached a peak value 3 weeks after the onset of hepatitis, whereas seroprevalence of GL IgG and YES IgG peaked after the disease had subsided. E2 IgG persisted in all animals for the entire duration of the experiment of up to 86 weeks and possibly beyond and, thus, can serve as a useful epidemiological marker of HEV infection. PMID- 14556265 TI - Frequency of human cytomegalovirus-specific T cells during pregnancy determined by intracellular cytokine staining. AB - The characteristics of cytomegalovirus (CMV)-specific T-cell immunity was investigated in pregnant women with primary, latent, or reactivated CMV infection, and in a comparative group of non-pregnant women. Forty-six pregnant and 8 non-pregnant women were examined based on the presence of serum antibody activity against CMV and viral excretion in urine. The frequency of CMV-specific CD4(+) T cells in peripheral blood lymphocytes was determined by staining for intracellular cytokines, interferon (IFN)-gamma, and tumor necrosis factor (TNF) alpha. There was no change in the frequencies of CMV-specific CD4(+) T cells in CMV-seropositive normal non-pregnant and pregnant women at any gestation. However, the frequency of CMV-specific CD4(+) T cells in pregnant women associated with CMV reactivation or reinfection was significantly higher than in CMV-seropositive normal pregnant and non-pregnant women. There were no CMV transmissions to the infants of all these women. These CMV-specific T cells responses in pregnant women may contribute some to block the intrauterine CMV infection in their infants. PMID- 14556266 TI - Behavior of the dengue virus in solution. AB - The dengue virus consists of four antigenically related but distinct viruses, termed Dengue virus 1-4 (DEN 1-4). We have established that the dengue virus loses infectivity over time in solution in an exponentially declining manner. The four strains examined (one from each serotype) have half-lives that range from 2.5 to 7.5 hr in defined medium. The half-life is temperature and pH-dependent and is affected by the nature of the host cell in which it is produced, but is not dependent upon the presence of either Mg(2+) ions or chelating agents. Electron microscopy (EM) of solutions of the dengue virus show almost complete virus aggregation after 24 hr at room temperature, while RT-PCR shows an intact RNA genome. These results show that the solution environment of the dengue virus is an important determinant of dengue virus infectivity. PMID- 14556267 TI - Detection of enterovirus in human skeletal muscle from patients with chronic inflammatory muscle disease or fibromyalgia and healthy subjects. AB - Enterovirus RNA has been found previously in specimens of muscle biopsy from patients with idiopathic dilated cardiomyopathy, chronic inflammatory muscle diseases, and fibromyalgia or chronic fatigue syndrome (fibromyalgia/chronic fatigue syndrome). These results suggest that skeletal muscle may host enteroviral persistent infection. To test this hypothesis, we investigated by reverse transcription-polymerase chain reaction (RT-PCR) assay the presence of enterovirus in skeletal muscle of patients with chronic inflammatory muscle diseases or fibromyalgia/chronic fatigue syndrome, and also of healthy subjects. Three of 15 (20%) patients with chronic inflammatory muscle diseases, 4 of 30 (13%) patients with fibromyalgia/chronic fatigue syndrome, and none of 29 healthy subjects was found positive. The presence of VP-1 enteroviral capsid protein was assessed by an immunostaining technique using the 5-D8/1 monoclonal antibody; no biopsy muscle from any patient or healthy subject was found positive. The presence of viral RNA in some muscle biopsies from patients exhibiting muscle disease, together with the absence of VP-1 protein, is in favor of a persistent infection involving defective viral replication. PMID- 14556268 TI - Molecular epidemiology of Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 strains from Russian patients with classic, posttransplant, and AIDS-associated Kaposi's sarcoma. AB - We report the molecular characterization of 38 new Kaposi's sarcoma-associated herpesvirus (KSHV) strains from Russian patients with either classic (25 cases), epidemic/AIDS-associated (7 cases), or posttransplant/immunosuppressed patients (6 cases), or Kaposi's sarcoma (KS). While a complete sequence of the K1 gene (870 bp) was obtained from 30 strains, only partial sequences of the hypervariable regions VR1 (372 bp) and/or VR2 (381 bp) of the K1 gene were obtained from eight strains of KS paraffin blocks. Sequence comparison and phylogenetic studies indicate that the novel KSHV strains belong to either the A subtype (28 cases) or the C subtype (10 cases). Within the 28 strains of A subtype, 24 (86%) belong to the large A' subgroup, mostly A1 and A1' clades, and 4 belong to the A" subgroup, mostly A3 clade. Within the 10 strains of subtype C, 4 were of C' subgroup, and 6 of the C". Some molecular variants of subtype A' were observed, with 3 strains exhibiting an insertion of a single amino acid at the position 65 and 2 strains (both from AIDS-KS) with an unique deletion of 17 amino acids in the VR2 region. Polymerase chain reaction-based subtyping of the K14.1 genomic region indicated that most (23/32) of the novel strains belonged to the P subtype. The results indicate that despite a wide genetic diversity of A and C K1 subtypes of KSHV strains present in Russia, most are closely related and belong to the A1 or A1' molecular clades suggesting a common origin. This study also expands the data regarding the absence of any correlation between a K1 molecular subtype and a specific KS type (classic, epidemic, or posttransplant), as well as between the K1 and K14.1 molecular subtypes. PMID- 14556269 TI - Real-time nested multiplex PCR for the detection of herpes simplex virus types 1 and 2 and varicella zoster virus. AB - One hundred forty-nine specimens were tested in a LightCycler nested multiplex polymerase chain reaction (LCnmPCR) for Herpes simplex virus (HSV)1, HSV2, and VZV. Eighty-one were from genitourinary medicine (GUM) patients and the other 68 specimens were from other patients with skin lesions. The results were compared to a conventional multiplex nested PCR (nmPCR) using agarose gel electrophoresis. Twenty-five specimens were positive in both assays for HSV1 and 29 were positive for VZV. For HSV2 there were 27 positive in the LCnmPCR and 26 positive in the nmPCR assay. The melting temperatures (Tms) of each target were different with a mean of 84.75 degrees C for HSV1, 88.57 degrees C for HSV2, and 83.62 degrees C for VZV. The melting curves of positive specimens directly overlaid the melting curves of the positive controls in the assay. The LCnmPCR assay is a convenient alternative to conventional PCR using agarose gel electrophoresis. It improves specimen turnaround time by eliminating the need for gel electrophoresis, transillumination, and gel photography. It also shows increased sensitivity for HSV2 over our standard assay. This LCnmPCR reduces further the possibility of amplicon contamination with nested PCR protocols. PMID- 14556270 TI - Parameters of CpG oligodeoxynucleotide-induced protection against intravaginal HSV-2 challenge. AB - Synthetic oligodeoxynucleotides (ODN) containing unmethylated CpG dinucleotides within the context of certain flanking bases (CpG motifs) have been shown to induce potent innate and adaptive immune responses. Vaginal delivery of CpG ODN alone protects mice from vaginal herpes simplex virus type-2 (HSV-2) challenge. Here, we investigated the importance of timing of delivery, formulation, route and dose of vaginally administered CpG ODN in the prevention or treatment of intravaginal (IVAG) HSV-2 infection. Mice treated intravaginally with CpG ODN containing a phosphorothioate backbone 24 hours prior to IVAG HSV-2 challenge survived infection, showed minimal vaginal pathology, and had virtually no detectable virus in vaginal washes, when compared to mice treated with non-CpG ODN. Genital treatment of HSV-2 infected mice with CpG ODN 4 hours after infection resulted in increased survival and decreased pathology and vaginal virus titers, whereas treatment of infected mice with CpG ODN 24 and 72 hours after IVAG HSV-2 infection had no effect on disease progression. Both liquid and solid (delivered on a bio-erodible muco-adhesive film) formulations of CpG ODN were effective in protection against genital HSV-2 following vaginal delivery. Lastly, IVAG delivery of 10 micro g of CpG ODN protected as well as a 100 micro g dose. PMID- 14556271 TI - Mathematical model to simulate the cellular dynamics of infection with human herpesvirus-6 in EBV-negative infectious mononucleosis. AB - Acute infection with human herpesvirus-6 induces physiological cell proliferation in persons without major immune deficiency. It thus can serve as a parameter to validate a mathematical model designed to simulate cell proliferation under physiological and pathological conditions. Such a mathematical model is presented to simulate various cell changes of the T-cell immune system during the course of HHV-6 infection. Model development follows several steps, beginning with a basic model containing physiological T-cell pools to the introduction of infectious stimuli in the final model. A search algorithm designed to optimize the system parameters, as well as initial variables of the model, is presented. The results of simulation runs for acute HHV-6 infection of the final computational model correspond well to the data, as documented in human patients; they suggest that the computational model presented for the simulation of T-cell levels in a given viral infection may well serve as a tool for similar studies of other viral infections, including those that lead to cellular aplasia or neoplasia. PMID- 14556272 TI - Analysis of human herpesvirus-6 IE1 sequence variation in clinical samples. AB - Herpesvirus immediate early (IE) proteins are known to play key roles in establishing productive infections, regulating reactivation from latency, and creating a cellular environment favourable to viral replication. Human herpesvirus-6 (HHV-6) IE genes have not been studied as intensively as their homologues in the prototype betaherpesvirus human cytomegalovirus (HCMV). Whilst the HCMV IE1 gene is relatively conserved, early studies indicated that HHV-6 IE1 exhibited a high level of sequence variation between HHV-6A and HHV-6B isolates, although the observation was based primarily on virus stocks that had been isolated and propagated in vitro. In this study, we investigated the level of HHV 6 IE1 sequence variation in vivo by direct sequencing of circulating virus in clinical samples without prior in vitro culture. Sequences exactly matching those reported for reference HHV-6 isolates were identified in clinical samples, thus the HHV-6 laboratory strains used in the majority of in vitro studies appear to be representative of virus circulating in vivo with respect to the IE1 gene. The HHV-6 IE1 sequence is also conserved in reference strains that had been passaged extensively in vitro. The high degree of divergence between variant A and B type IE1 sequences was confirmed, but interestingly HHV-6B IE1 sequences were observed to further segregate into two distinct subgroups, with the laboratory strains Z29 and HST representative of these two subgroups. Within each HHV-6B subgroup, a remarkably high level of homology was observed. Thus the HHV-6 IE1 sequence appears highly stable, underlining its potential importance to the viral life cycle. PMID- 14556273 TI - Moderate variation of the oncogenic potential among high-risk human papillomavirus types in gynecologic patients with cervical abnormalities. AB - The oncogenic potential of human papillomavirus (HPV) infection was assessed by following the disease course in 455 patients who had had a routine diagnostic Hybrid Capture HPV test due to squamous cell abnormalities of the uterine cervix as detected by cytology and/or colposcopy. At entry, 308 patients had cytologic atypia classified as P3 by the Papanicolau classification, 168 had a positive high-risk HPV test, and 23 were infected only with low-risk HPV. The patients were followed-up using the patient registry until the endpoint of histologically diagnosed cervical intraepithelial neoplasia (CIN). High-grade CIN was diagnosed in 75 surgical biopsies. High-risk HPV infection (relative risk: 76.8 CI(95): 23.7-249.5), cytologic atypia (RR: 16.2 CI(95): 3.9-66.6), and age above 35 (RR: 1.99 CI(95): 1.26-3.16) were independent risk factors for high-grade CIN, while the viral load did not predict oncogenic progression (P = 0.47). After PCR-RFLP typing, the high-risk types were classified into groups as follows: (1) types 16 and 18, (2) types 45, 52, and 56, (3) types 31, 33, 35, 51, and 58. The relative risks of high-grade CIN were 119.1 (CI(95): 36.2-390.9) for group 1, 44.4 (CI(95): 9.8-201) for group 2, and 39.7 (CI(95): 10.9-144.8) for group 3, respectively. The risk ratios between the groups of high-risk types were found to differ at most by a factor of 2.98 (corrected P value: 0.007) indicating that the oncogenic potential varies moderately within the high-risk group of HPVs. PMID- 14556274 TI - Buccal exposure to human papillomavirus type 16 is a common yet transitory event of childhood. AB - High-risk human papillomaviruses, such as type 16 (HPV-16), are established etiological agents for cervical carcinoma. In most cases, this virus is transmitted sexually, though can also be spread from mother to infant at delivery. We have demonstrated previously a high prevalence ( approximately 52%) of HPV-16 DNA in the mouths of prepubertal children, albeit with low levels of transcription [Rice et al., 2000]. We investigated whether childhood buccal infections with HPV-16 are persistent or transient and whether children became infected through contact with their immediate family members. Two groups of children were selected: one group were all initially HPV-16 E5 DNA-positive in sensitive polymerase chain reaction tests of swabs from their buccal mucosa (n = 20), and the other group consisted of children who were all HPV-16 E5-negative (n = 19). Thirty months later, a second oral swab was collected from each child and tested for HPV DNA. At this second visit, 40% of the HPV-16-positive group had no detectable HPV-16 DNA; conversely, 63% of children who were originally HPV-16 negative had now acquired the virus. Three months later, a third sample was collected from eight children and their immediate families (seven were HPV-16 E5 DNA-positive at the second visit). Amongst the family samples tested, in two families a single previously untested child was HPV-16 DNA-positive. It is concluded that HPV-16 DNA in the oral cavities of children is a transient event and is most probably acquired from their peers. PMID- 14556275 TI - Molecular epidemiology of measles virus in Kenya. AB - Measles causes significant morbidity and mortality globally. Many countries have embarked on immunization programs to control and prevent measles outbreaks and eventually to eliminate endemic measles. Kenya is currently in the outbreak control and prevention stage for measles. Measles virus genotyping is important for molecular epidemiological purposes, including the documentation of the elimination of endemic measles virus strains from a country, and mapping of transmission pathways. In this study, we collected clinical specimens from measles outbreak cases in 2002 in Kenya for measles virus genotyping. We were able to isolate and/or detect measles virus in 10 cases from 5 of the 8 provinces in Kenya. All these Kenyan measles strains were determined to be genotype D4 strains when compared to the standard World Health Organization-designated measles virus reference strains. Interestingly, the Kenyan D4 strains clustered into two distinct D4 subgroups. In addition, the inclusion of other published D4 measles strains in this analysis indicated that there are four distinct D4 clusterings, or subgroups: Montreal-like, India-like, Johannesburg-like, and Ethiopia-like. This is the first measles molecular epidemiology study in Kenya and establishes the current endemic measles strain as genotype D4. Importantly, this study shows that the Kenyan D4 strains are distinct from the B3 measles strain found in West Africa and the D4 strains reported in Ethiopia. PMID- 14556276 TI - Parvovirus B19 infection in Taiwanese patients with hematological disorders. AB - Human parvovirus B19 has a strong tissue tropism for erythroid progenitor cells and is a causative agent for anemia. However, it remains unclear whether patients with hematological disorders are at a higher risk of B19 infection. In the present study, we evaluated the prevalence of B19 infection in 278 patients with hematological disorders by indirect antibody capture enzyme-linked immunosorbent assay. Virus inoculation into cell culture (of TF-6, WRL-68, and HL-60) was carried out using serum from patients with positive IgM anti-B19 and was then examined by nested polymerase chain reaction, dot-blot analysis, and sequence analysis. Our data demonstrated that the total seropositive rates of either IgG or IgM were 71.9%. The seropositive rates increase significantly with age (P < 0.001). After adjustment for age, the seropositive rate was significantly higher in our patients than in the general population with standardized rate ratio of 1.56 (95% CI = 1.43-1.68). No significant difference was found among different disease subgroups (P = 0.311). Nine patients (3.2%) had active B19 infection with positive IgM antibody, with four diagnosed as having idiopathic thrombocytopenic purpura (ITP). Viremia of B19 virus could be detected in eight of nine patients, including three patients in serum only, three patients in bone marrow only, and two patients in both serum and bone marrow. We conclude that patients with hematological disease have higher seropositive rates for B19 than occur in normal controls and that study of occult parvovirus B19 infection is recommended in patients with hematological disease. PMID- 14556277 TI - Characterisation of human tick-borne encephalitis virus from Sweden. AB - Viruses of the tick-borne encephalitis (TBE) antigenic complex, within the family Flaviviridae, cause a variety of diseases including uncomplicated febrile illness, meningo-encephalitis and haemorrhagic fever. Different wildlife species act as reservoir hosts with ixodid tick species as vectors. TBE virus (TBEV) causes 40-130 cases confirmed serologically in Sweden each year. Characteristics of TBEV strains circulating in Sweden have not been investigated previously and no viral sequence data has been reported. In the present study, virus strains were isolated from serum of patients with clinical symptoms consistent with acute TBEV infection. Serologic characterisation, using a panel of E-specific monoclonal antibodies and cross-neutralisation tests, indicated that the Swedish strains of TBEV, isolated 1958-1994, all belonged to the Western TBEV subtype, which includes the Austrian vaccine strain Neudoerfl. Genetic analysis of a partial E-sequence confirmed this close relationship: all Swedish TBEV strains belonged to the European lineage of the Western TBEV subtype, which includes the previously characterised strains Neudoerfl, Hypr, and Kumlinge. Further, three Swedish strains showed partial E-sequences identical to that of the Finnish Kumlinge strain, ten Swedish strains formed a well-supported separate cluster, whereas four others did not show any real clustering. No apparent correlation was observed in comparison of clinical parameters with genetic data or geographic origin of the strains. PMID- 14556278 TI - Seasonal changes in avian song control circuits do not cause seasonal changes in song discrimination in song sparrows. AB - In seasonally breeding songbirds, brain nuclei of the song control system that act in song perception change in size between seasons. It has been hypothesized that seasonal regression of song nuclei may impair song discrimination. We tested this hypothesis in song sparrows (Melospiza melodia), a species in which males share song types with neighbors and must discriminate between similar songs in territorial interactions. We predicted that song sparrows with regressed song systems would have greater difficulty in discriminating between similar songs. Sparrows were housed either on short days (SD) and had regressed song circuits, or were exposed to long days and implanted with testosterone (LD+T) to induce full growth of the song circuits. We conducted two experiments using a GO/NO-GO operant conditioning paradigm to measure song discrimination ability of each group. Birds learned four (experiment 1) or three (experiment 2) pairs of song types sequentially, with each pair more similar in the number of shared song elements and thus more difficult to discriminate. Circulating T levels differed between the SD and LD+T groups. The telencephalic song nuclei HVc, RA, and area X were larger in the LD+T birds. The two groups of sparrows did not differ, however, in their ability to learn to discriminate between shared song types, regardless of the songs' similarity. These results suggest that seasonal changes in the song control system do not affect birds' ability to make difficult song discriminations. PMID- 14556279 TI - Androgens and estrogens induce seasonal-like growth of song nuclei in the adult songbird brain. AB - In seasonally breeding songbirds, the brain regions that control song behavior undergo dramatic structural changes at the onset of each annual breeding season. As spring approaches and days get longer, gonadal testosterone (T) secretion increases and triggers the growth of several song control nuclei. T can be converted to androgenic and estrogenic metabolites by enzymes expressed in the brain. This opens the possibility that the effects of T may be mediated via the androgen receptor, the estrogen receptor, or both. To test this hypothesis, we examined the effects of two bioactive T metabolites on song nucleus growth and song behavior in adult male white-crowned sparrows. Castrated sparrows with regressed song control nuclei were implanted with silastic capsules containing either crystalline T, 5alpha-dihydrotestosterone (DHT), estradiol (E(2)), or a combination of DHT+E(2). Control animals received empty implants. Song production was highly variable within treatment groups. Only one of seven birds treated with E(2) alone was observed singing, whereas a majority of birds with T or DHT sang. After 37 days of exposure to sex steroids, we measured the volumes of the forebrain song nucleus HVc, the robust nucleus of the archistriatum (RA), and a basal ganglia homolog (area X). All three steroid treatments increased the volumes of these three song nuclei when compared to blank-implanted controls. These data demonstrate that androgen and estrogen receptor binding are sufficient to trigger seasonal song nucleus growth. These data also suggest that T's effects on seasonal song nucleus growth may depend, in part, upon enzymatic conversion of T to bioactive metabolites. PMID- 14556281 TI - Neural correlates to flight-related density-dependent phase characteristics in locusts. AB - Locust phase polymorphism is an extreme example of behavioral plasticity; in response to changes in population density, locusts dramatically alter their behavior. These changes in behavior facilitate the appearance of various morphological and physiological phase characteristics. One of the principal behavioral changes is the more intense flight behavior and improved flight performance of gregarious locusts compared to solitary ones. Surprisingly, the neurophysiological basis of the behavioral phase characteristics has received little attention. Here we present density-dependent differences in flight-related sensory and central neural elements in the desert locust. Using techniques already established for gregarious locusts, we compared the response of locusts of both phases to controlled wind stimuli. Gregarious locusts demonstrated a lower threshold for wind-induced flight initiation. Wind-induced spiking activity in the locust tritocerebral commissure giants (TCG, a pair of identified interneurons that relay input from head hair receptors to thoracic motor centers) was found to be weaker in solitary locusts compared to gregarious ones. The solitary locusts' TCG also demonstrated much stronger spike frequency adaptation in response to wind stimuli. Although the number of forehead wind sensitive hairs was found to be larger in solitary locusts, the stimuli conveyed to their flight motor centers were weaker. The tritocerebral commissure dwarf (TCD) is an inhibitory flight-related interneuron in the locust that responds to light stimuli. An increase in TCD spontaneous activity in dark conditions was significantly stronger in gregarious locusts than in solitary ones. Thus, phase dependent differences in the activity of flight-related interneurons reflect behavioral phase characteristics. PMID- 14556280 TI - Limits on volume changes in the mushroom bodies of the honey bee brain. AB - The behavioral maturation of adult worker honey bees is influenced by a rising titer of juvenile hormone (JH), and is temporally correlated with an increase in the volume of the neuropil of the mushroom bodies, a brain region involved in learning and memory. We explored the stability of this neuropil expansion and its possible dependence on JH. We studied the volume of the mushroom bodies in adult bees deprived of JH by surgical removal of the source glands, the corpora allata. We also asked if the neuropil expansion detected in foragers persists when bees no longer engage in foraging, either because of the onset of winter or because colony social structure was experimentally manipulated to cause some bees to revert from foraging to tending brood (nursing). Results show that adult exposure to JH is not necessary for growth of the mushroom body neuropil, and that the volume of the mushroom body neuropil in adult bees is not reduced if foraging stops. These results are interpreted in the context of a qualitative model that posits that mushroom body neuropil volume enlargement in the honey bee has both experience-independent and experience-dependent components. PMID- 14556282 TI - Dehydroepiandrosterone upregulates neural androgen receptor level and transcriptional activity. AB - The mechanism of action of dehydroepiandrosterone (DHEA), a neuroactive neurosteroid synthesized in the brains of humans and other mammals, has not been fully characterized in the adult brain. Although well known for modulatory effects on GABA(A), NMDA, and sigma(1) receptors, studies in both CNS and peripheral target cells suggest that DHEA also may exert genomic effects via the androgen receptor (AR). The current study tested the hypothesis that DHEA was capable of producing androgenic effects in the CNS by assaying its ability to induce three characteristic effects of an androgenic compound. These included the ability to upregulate neural AR protein level in mouse brain and immortalized GT1 7 hypothalamic cells, the capacity to induce transcriptional activity through AR in CV-1 cells transfected with an MMTV-ARE-CAT reporter, and competition for recombinant AR binding in a radioligand binding assay. The results showed that DHEA treatment significantly augmented AR both in vivo and in vitro, and that this effect was not blocked by trilostane (TRIL), a known 3beta-hydroxysteroid dehydrogenase (3beta-HSD) inhibitor. DHEA also promoted AR-mediated CAT reporter expression and competed with dihydrotestosterone (DHT) for binding to recombinant AR in a cell-free system. These data indicate that DHEA possesses intrinsic androgenic activity that is potentially independent of metabolic conversion to other androgens, and that it can affect gene function through the AR. In combination with its modulation of neurotransmitter receptors at the cell membrane level, the findings suggest that the mechanism of action of DHEA in the brain can involve a "crosstalk" cellular signaling system that involves both nongenomic and genomic components. PMID- 14556283 TI - Balance of cell proliferation and death among dynamic populations: a mathematical model. AB - Developmental changes in cell numbers represent the dynamic balance between cell proliferation and death. One obstacle to assessing this balance is an inability to quantify the total amount of cell death, i.e., with a positive indicator such as terminal dUTP nick end labeling (TUNEL) or caspase activity. A novel mathematical model is described wherein data on daily cell growth (the change in cell number) and cell cycle kinetics can be used to determine the total amount of cell death. Two sets of data from previously published studies were tested in this model; primary cultured cortical neurons and B104 neuroblastoma cells. These two preparations have contrasting features: neuronal cultures are heterogeneous and have relatively few cells that are actively cycling (i.e., the growth fraction for these cells is low), whereas B104 cells are relatively homogeneous cultures in which the growth fraction is high. In primary cortical cultures, there was a balance in cell production and death. Treatment with a potent anti mitogen, ethanol (400 mg/dl), affected this balance principally by reducing cell production, although the rate of cell death was also increased. In untreated B104 cells, there was eight-fold more cell production than cell death. Growth factors such as platelet-derived growth factor BB doubled cell production. Ethanol reduced cell production by >60%, and it eliminated growth factor-mediated cell production. All of these changes occurred in the absence of an effect on the amount of cell death. Thus, the model is ideal for predicting the effects of an epigenetic factor (e.g., a growth factor, toxin, or pharmacological agent) on cell development and can be useful in determining the consequences of a genetic manipulation as well. PMID- 14556284 TI - Methylene blue blocks cGMP production and disrupts directed migration of microglia to nerve lesions in the leech CNS. AB - Migration and accumulation of microglial cells at sites of injury are important for nerve repair. Recent studies on the leech central nervous system (CNS), in which synapse regeneration is successful, have shown that nitric oxide (NO) generated immediately after injury by endothelial nitric oxide synthase (eNOS) stops migrating microglia at the lesion. The present study obtained results indicating that NO may act earlier, on microglia migration, and aimed to determine mechanisms underlying NO's effects. Injury induced cGMP immunoreactivity at the lesion in a pattern similar to that of eNOS activity, immunoreactivity, and microglial cell accumulation, which were all focused there. The soluble guanylate cyclase (sGC) inhibitor methylene blue (MB) at 60 microM abolished cGMP immunoreactivity at lesions and blocked microglial cell migration and accumulation without interfering with axon conduction. Time-lapse video microscopy of microglia in living nerve cords showed MB did not reduce cell movement but reduced directed movement, with significantly more cells moving away from the lesion or reversing direction and fewer cells moving toward the lesion. The results indicate a new role for NO, directing the microglial cell migration as well as stopping it, and show that NO's action may be mediated by cGMP. PMID- 14556285 TI - Specification of motoneuron fate in Drosophila: integration of positive and negative transcription factor inputs by a minimal eve enhancer. AB - We are interested in the mechanisms that generate neuronal diversity within the Drosophila central nervous system (CNS), and in particular in the development of a single identified motoneuron called RP2. Expression of the homeodomain transcription factor Even-skipped (Eve) is required for RP2 to establish proper connectivity with its muscle target. Here we investigate the mechanisms by which eve is specifically expressed within the RP2 motoneuron lineage. Within the NB4-2 lineage, expression of eve first occurs in the precursor of RP2, called GMC4-2a. We identify a small 500 base pair eve enhancer that mediates eve expression in GMC4-2a. We show that four different transcription factors (Prospero, Huckebein, Fushi tarazu, and Pdm1) are all expressed in GMC4-2a, and are required to activate eve via this minimal enhancer, and that one transcription factor (Klumpfuss) represses eve expression via this element. All four positively acting transcription factors act independently, regulating eve but not each other. Thus, the eve enhancer integrates multiple positive and negative transcription factor inputs to restrict eve expression to a single precursor cell (GMC4-2a) and its RP2 motoneuron progeny. PMID- 14556286 TI - Rapid electrical and delayed molecular signals regulate the serum response element after nerve injury: convergence of injury and learning signals. AB - Axotomy elicits changes in gene expression, but little is known about how information from the site of injury is communicated to the cell nucleus. We crushed nerves in Aplysia californica and the sciatic nerve in the mouse and found short- and long-term activation of an Elk1-SRF transcription complex that binds to the serum response element (SRE). The enhanced short-term binding appeared rapidly and was attributed to the injury-induced activation of an Elk1 kinase that phosphorylates Elk1 at ser383. This kinase is the previously described Aplysia (ap) ERK2 homologue, apMAPK. Nerve crush evoked action potentials that propagated along the axon to the cell soma. Exposing axons to medium containing high K(+), which evoked a similar burst of spikes, or bathing the ganglia in 20 microM serotonin (5HT) for 20 min, activated the apMAPK and enhanced SRE binding. Since 5HT is released in response to electrical activity, our data indicate that the short-term process is initiated by an injury-induced electrical discharge that causes the release of 5HT which activates apMAPK. 5HT is also released in response to noxious stimuli for aversive learning. Hence, apMAPK is a point of convergence for injury signals and learning signals. The delay before the onset of the long-term SRE binding was reduced when the crush was closer to the ganglion and was attributed to an Elk1 kinase that is activated by injury in the axon and retrogradely transported to the cell body. Although this Elk1 kinase phosphorylates mammalian rElk1 at ser383, it is distinct from apMAPK. PMID- 14556287 TI - Neuromuscular activity impairs axonal sprouting in partially denervated muscles by inhibiting bridge formation of perisynaptic Schwann cells. AB - Following partial denervation of rat hindlimb muscle, terminal Schwann cells extend processes from denervated endplates to induce and guide sprouting from the remaining intact axons. Increased neuromuscular activity significantly reduces motor unit enlargement and sprouting during the acute phase of sprouting. These findings led to the hypothesis that increased neuromuscular activity perturbs formation of Schwann cell bridges and thereby reduces sprouting. Adult rat tibialis anterior (TA) muscles were extensively denervated by avulsion of L4 spinal root and were immediately subjected to normal caged activity or running exercise (8 h daily) for 3, 7, 14, 21, and 28 days. Combined silver/cholinesterase histochemical staining revealed that the progressive reinnervation of denervated endplates by sprouts over a 1 month period in the extensively partially denervated TA muscles was completely abolished by increased neuromuscular activity. Immunohistochemical staining and triple immunofluorescence revealed that the increased neuromuscular activity did not perturb the production of Schwann cell processes, but prevented bridging between Schwann cell processes at innervated and denervated endplates. Our findings suggest that failure of Schwann cell processes to bridge between endplates accounts, at least in part, for the inhibitory effect of increased neuromuscular activity on sprouting. PMID- 14556288 TI - Synaptic connections of cholinergic antennal lobe relay neurons innervating the lateral horn neuropile in the brain of Drosophila melanogaster. AB - Presumed cholinergic projection neurons (PNs) in the brain of the fruit fly Drosophila melanogaster, immunoreactive to choline acetyltransferase (ChAT), convey olfactory information between the primary sensory antennal lobe neuropile and the mushroom body calyces, and finally terminate in the lateral horn (LH) neuropile. The texture and synaptic connections of ChAT PNs in the LH and, comparatively, in the smaller mushroom body calyces were investigated by immuno light and electron microscopy. The ChAT PN fibers of the massive inner antennocerebral tract (iACT) extend into all portions of the LH, distributing in a nonrandom fashion. Immunoreactive boutons accumulate in the lateral margins of the LH, whereas the more proximal LH exhibits less intense immunolabeling. Boutons with divergent presynaptic sites, unlabeled as well as ChAT immunoreactive, appear to be the preponderant mode of synaptic input throughout the LH. Synapses of ChAT-labeled fibers appear predominantly as divergent synaptic boutons (diameters 1-3 microm), connected to unlabeled postsynaptic profiles, or alternatively as a minority of tiny postsynaptic spines (diameters 0.05-0.5 microm) among unlabeled profiles. Together these spines encircle unidentified presynaptic boutons of interneurons which occupy large areas of the LH. Thus, synaptic circuits in the LH differ profoundly from those of the PNs in the mushroom body calyx, where ChAT spines have not been encountered. Synaptic contacts between LH ChAT elements were not observed. The synaptic LH neuropile may serve as an output area for terminals of the ChAT PNs, their presynaptic boutons providing input to noncholinergic relay neurons. The significance of the postsynaptic neurites of the ChAT PNs is discussed; either local or other interneurons might connect the ChAT PNs within the LH, or PNs might receive inputs arising from outside the LH. PMID- 14556289 TI - Origin of the isthmus? A comparison of the brains of lancelets and vertebrates. PMID- 14556290 TI - Cytoarchitecture, topography, and descending supraspinal projections in the anterior central nervous system of Branchiostoma lanceolatum. AB - The central nervous system (CNS) of the chordate amphioxus (Branchiostoma lanceolatum) is divisible into a spinal cord and an anterior portion in some ways equivalent to the brain of craniates. The present study reports on this anterior portion, with respect to general topography, cytoarchitecture, and cells that give rise to descending supraspinal projections. The anterior portion of the CNS is located adjacent to the first four myomeres and rostral to the first giant cell of Rohde-it can be divided into several regions that differ with respect to their cytoarchitecture. The tip of the neural tube is formed by a small anterior vesicle; caudally, there is a much larger region that is intercalated between the anterior vesicle and the first cell of Rohde. This intercalated region, in turn, consists of three subdivisions: an anterior subdivision adjacent to myomere 1, an intermediate subdivision adjacent to myomere 2, and a posterior one adjacent to myomeres 3 and 4. After injections of tracers into the spinal cord a large number of cells were labeled in the intercalated region. The spinally projecting cells were not evenly distributed: their number was decreased in the center of the intermediate subdivision. These subdivisions, which have previously not been noted, may be aligned with the expression domains of regulatory genes (e.g., AmphiOtx, AmphiHox) in larval lancelets. In particular, the center of the intermediate subdivision may correspond to a "nonHox/nonOtx" domain in the CNS of the larva. A similar embryonic domain occurs in the brain of craniates in which it develops into the isthmus cerebri that separates mid- and hindbrain. A close structural and topographical inspection of the corresponding region of adult lancelets reveals, however, that this region is not the homolog of an isthmus, but a uniquely derived, autapomorphic feature of lancelets. PMID- 14556291 TI - Internalization of interphotoreceptor retinoid-binding protein by the Xenopus retinal pigment epithelium. AB - Xenopus rods and cones secrete into the interphotoreceptor matrix (IPM) a 124-kDa glycoprotein termed interphotoreceptor retinoid-binding protein (IRBP; Hessler et al. [1996] J. Comp. Neurol. 367:329-341). IRBP is confined to the IPM, being too large to diffuse through the zonulae adherentes between adjacent photoreceptor and Muller cells. Despite this physical entrapment within the subretinal space, IRBP is rapidly cleared from the IPM by an unknown mechanism. Immunohistochemistry and immunoelectron microscopy were used to localize IRBP in intact and detached retina-retinal pigment epithelium (RPE) eyecups. The effects of light, dark, and time of day on the compartmentalization of IRBP were characterized by quantitative Western blot analysis and by immunoprecipitation of IRBP labeled in vivo by intraocular injection of [(35)S]methionine. Immunohistochemistry showed that the apparent intercellular IRBP in both the RPE and the photoreceptors is resistant to saline extraction, in contrast to that in the IPM. In the RPE, IRBP was associated with matrix material within phagosomes and endosomes. The IPM, RPE, and retina contained 75%, 18%, and 7% of the total IRBP in the eye, respectively. The IPM and RPE contain 130 +/- 14 pmoles and 34 +/- 4 pmoles of IRBP, respectively. The amounts of IRBP in the RPE at middark and midlight were the same. Furthermore, the in vivo uptake of [(35)S]methionine labeled IRBP was light independent. Our studies suggest that IRBP is not strictly confined to the subretinal space but rather that significant amounts are present intracellularly, particularly within the RPE, which does not synthesize IRBP. Furthermore, IRBP secreted by the photoreceptors is taken up from the IPM mainly through a light-independent endocytic pathway separate from outer segment phagocytosis. The role of RPE endocytosis should be explored in relation to the function of IRBP. PMID- 14556292 TI - Dopaminergic amacrine cells in the inner nuclear layer and ganglion cell layer comprise a single functional retinal mosaic. AB - Many types of retinal neuron are distributed in an orderly manner across the surface of the retina. Indeed, the existence of such regularity amongst a population of neurons, termed a retinal mosaic, may be a defining feature of functionally independent types of retinal neuron. We have examined the spatial distribution of dopaminergic amacrine cells in the ferret retina both in the inner nuclear layer (INL) and in the ganglion cell layer (GCL) to determine whether the cells in each layer form an independent retinal mosaic as evidence of whether they should be considered as two separate types. Ferret retinas contain approximately 1,900 dopaminergic amacrine cells, of which 27% are located in the GCL, and the rest in the INL. Based on analysis of their Voronoi domains as well as autocorrelation analysis and tests for complete spatial randomness, we found that the distribution of INL cells was statistically regular, while that of the GCL cells was not. However, by using cross-correlation analysis, these two groups of cells were found to be spatially dependent: an exclusion zone was detected in the cross-correlogram of roughly the same size as that found in the autocorrelograms of both INL and GCL cells. Such a pattern would be expected if dopaminergic amacrine cells in the INL and GCL were members of a single regular population differing only in their somatic depth. By using computer simulations, we tested this hypothesis directly, confirming that a random assignment of 27% from the total population produces cross-correlograms that are indistinguishable from those of the biological mosaics. We conclude, therefore, that the cells in the two layers form a single functional population; those in the GCL appear to be misplaced. Somatic positioning with respect to depth within the retina is not, by itself, a reliable guide for functional classification. PMID- 14556293 TI - Aging of the human cerebellum: a stereological study. AB - Cerebella from 19 normal Caucasian males, ages 19-84 years, were studied using stereological methods. Cerebellum was divided into four different regions: the anterior and posterior lobe, the vermis, and the flocculonodular lobe. Total volume of the cerebellar cortex and white matter, cerebellar surface area, total Purkinje and granule cell number, and the distribution of the volumes of the Purkinje cells and their nuclei were estimated in all four regions. The global white matter was reduced by 26% with age; the mean volume of the Purkinje cell body was decreased by 33% with no decrease in the volume of the Purkinje cell nuclei. A tendency towards a 16% total cerebellar volume loss was seen without a concomitant neuron loss. No global Purkinje or granule cell loss was detected with age, total Purkinje cell number being 28 x 10(6) (coefficient of variation, CV = 0.16) and total granule cell number 109 x 10(9) (CV = 0.17). However, a significant change was observed with age in the anterior lobe, where a selective 40% loss of both Purkinje and granule cells was found. Furthermore, a 30% loss of volume, mostly due to a cortical volume loss, was recorded in the anterior lobe, which is predominantly involved in motor control. PMID- 14556294 TI - Local commissural interneurons integrate information from intersegmental coordinating interneurons. AB - The information that coordinates movements of swimmerets on different segments of the crayfish abdomen is conducted by interneurons that originate in each abdominal ganglion. These interneurons project axons to neighboring ganglia and beyond. To discover the anatomy of these axons in their target ganglia, we used Neurobiotin and dextran-Texas Red microelectrodes to fill them near their targets. Coordinating axons coursed through these target ganglia close to the midline and extended only a few short branches that did not approach the lateral neuropils. Two of the three types of coordinating axons made direct synaptic connections with a class of local commissural interneurons that relayed the information to targets in the swimmeret pattern-generating circuits. These commissural interneurons, named here ComInt 1 neurons, followed a particular route to cross the midline and reach their targets. ComInt 1 neurons were nonspiking; they received EPSPs from the coordinating axons near the midline and transmitted this information to their targets in the lateral neuropils using graded transmission. The output of each ComInt 1 was restricted to a single local circuit and had opposite effects on the power-stroke and return-stroke motor neurons driven by that circuit. ComInt 1 neurons were direct postsynaptic targets of both descending and ascending coordinating axons that originated in other anterior and posterior ganglia. Because of phase differences in the impulses in these different coordinating axons, their signals arrived simultaneously at each ComInt 1. We discuss these findings in the context of alternative models of the intersegmental coordinating circuit. PMID- 14556295 TI - Consequences of interspecies antennal imaginal disc transplantation on organization of olfactory glomeruli and pheromone blend discrimination. AB - The antennal imaginal disc was transplanted between male larvae of two different heliothine moth species, Heliothis virescens (HV) and Helicoverpa zea (HZ). Males of these species respond to distinct pheromone blends, have different peripheral and central olfactory neuron specificities, as well as distinct arrangements of antennal lobe olfactory glomeruli, in the specialized male macroglomerular complex (MGC). After pupal development and adult eclosion, unilateral (with one antennal disc left intact) and bilateral antennal transplant males were assayed in a wind tunnel to both species' pheromone blends to determine their ability to discriminate between the two signals. The postmetamorphic developmental effects of interspecific transplantation upon the primary olfactory centers in the moth brain were then examined in these same individuals. Behavioral tests showed that both types of unilateral transplant continued to exhibit upwind anemotactic flight to the normal recipient blend with occasional flights to the donor blend. In contrast, bilateral transplants preferred the HV pheromone blend regardless of the direction of transplant, with some males of each type also responding to the HZ blend. Neuroanatomic evaluation of the MGC revealed that the donor arrangement of MGC glomeruli was induced in 73% HZ donor to HV recipient transplants and 56% of the reciprocal transplant. Surprisingly, several V-Z bilateral transplant males responded to both HV and HZ pheromone blends and had two HV MGC structures. This behavioral outcome was unexpected, because responses to the HV blend are mediated by inputs that are normally antagonistic to HZ males and the normal HV antenna lacks olfactory receptor neurons capable of responding to the essential minor pheromone component of the HZ blend. These data indicate a plasticity in developmental pathways regulating the expression of peripheral olfactory receptor neurons and in the glomerular processing of species-specific olfactory information. PMID- 14556296 TI - Hodological characterization of the medial amygdala in anuran amphibians. AB - Early studies in anuran amphibians defined the amygdala as a single unit that only later could be subdivided into medial and lateral parts with the achievement of sensitive immunohistochemical and tracing techniques. However, the terminology used was often misleading when comparing with "homologous" amygdaloid nuclei in amniotes. Recently, the basal telencephalon of anurans has been demonstrated to be more complex than previously thought, and distinct amygdaloid nuclei were proposed on the basis of immunohistochemistry. Moreover, developmental data are increasing that support this notion. In the present study, we analyzed the patterns of afferent and efferent connections of the medial amygdala (MeA; formerly amygdala pars lateralis), considered as the main target of the vomeronasal information from the accessory olfactory bulb, as in other vertebrates. By means of axonal transport of dextran amines, the afferent and efferent connections of the MeA were traced in Rana perezi and Xenopus laevis under in vivo and in vitro conditions. Largely similar results were found in both species. The results showed abundant intratelencephalic and extratelencephalic connections that were readily comparable to those of other tetrapods. Most of these connections were reciprocal and, in particular, the strong relation of the MeA with the hypothalamus, via the stria terminalis, was demonstrated. Immunohistochemical techniques showed staining patterns that revealed abundant peptidergic afferents to the MeA, as well as minor inputs containing other neurotransmitters such as catecholamines. Double-labeling experiments demonstrated that the peptidergic fibers that reach the MeA originate in the ventral hypothalamus, whereas the catecholaminergic innervation of the MeA arises in the caudal extent of the posterior tubercle. Taken together, the results about connectivity in our study support the comparison of the MeA in anurans with its counterparts (and similarly named) amygdaloid nuclei in amniotes. Most of the hodological features of the medial amygdala seem to be shared by those tetrapods with well-developed vomeronasal systems. PMID- 14556297 TI - Stereologic analysis of estrogen receptor alpha (ER alpha) expression in rat hypothalamus and its regulation by aging and estrogen. AB - The estrogen receptor alpha (ERalpha) in the hypothalamus plays important roles in the regulation of reproductive development, physiology, and behavior. However, the expression of the ERalpha may change during aging or in response to varying estrogen levels. The present study measured changes in the numbers of ERalpha expressing cells in specific hypothalamic and preoptic nuclei of ovariectomized female Sprague-Dawley rats at three ages (young [3-4 months], middle-aged [10-12 months], or old [24-26 months]) and with or without estrogen replacement. Numbers of ERalpha-immunoreactive neurons were quantified in four regions relevant to reproductive function: the anteroventral periventricular nucleus (AVPV), medial preoptic nucleus (MPN), arcuate nucleus (ARH), and ventromedial nucleus (VMN), using an unbiased stereologic approach. In the AVPV and VMN, significant age related increases in the numbers of ERalpha-expressing cells from the middle-aged to the old group were detected, and no differences were observed in the MPN and ARH, indicating that ERalpha neuron number is maintained or even elevated during aging. No significant effects of estrogen on ERalpha cell number were detected in any of the four regions studied. Therefore, ERalpha cell number in the rat hypothalamus and preoptic area changes with aging in a region-specific manner. PMID- 14556298 TI - Regeneration of supraspinal axons after complete transection of the thoracic spinal cord in neonatal opossums (Monodelphis domestica). AB - These studies define the time table and origin of supraspinal axons regenerating across a complete spinal transection in postnatal Monodelphis domestica. After lumbar (L1) spinal cord injection of fluorophore-dextran amine conjugate on postnatal (P) day 4, a consistent number of neurons could be labeled. The numbers of labeled neurons remained stable for several weeks, but subsequently declined by P60 in control animals and by P35 in animals with complete spinal transection (T4-T6) performed at P7. In control animals, 25-40% of neurons labeled with a fluorophore injected (L1) at P4 could also be double-labeled by a second fluorophore injected (T8-T10) at different older ages. In spinally transected animals, total numbers of neurons labeled with the second marker were initially lower compared with age-matched controls, but were not significantly different by 3 weeks after injury. The proportion of double-labeled neurons in spinally transected animals increased from approximately 2% 1 week after injury (P14) to approximately 50% by P60, indicating that a substantial proportion of neurons with axons transected at P7 is able to regenerate and persist into adulthood. However, the proportion of axons originating from regenerating neurons made only a small contribution at older ages to total numbers of fibers growing through the injury site, because much of development of the spinal cord occurs after P7. Evidence was obtained that degenerating neurons with both apoptotic and necrotic morphologies were present in brainstem nuclei; the number of neurons with necrotic morphology was much greater in the brainstem of animals with spinal cords transected at P7. PMID- 14556299 TI - Victims of our own success: will immunization remain the paradigm of effective prevention? Grantmakers In Health. AB - As part of its continuing mission to serve trustees and staff of health foundations and corporate giving programs, Grantmakers In Health (GIH) convened a select group of grantmakers and national experts who have made a major commitment to childhood immunization. The roundtable explored various factors influencing public acceptance of childhood immunization, with discussions ultimately centering on the importance of ensuring and conveying accurate information to the public and policymakers. Current and potential roles for health philanthropy were also discussed. This report brings together key points from the day's discussion with factual information on childhood immunization drawn from a background paper prepared for the meeting. When available, recent findings, facts, and figures have been incorporated. PMID- 14556300 TI - Grassroots health care coverage: local communities are trying novel approaches to providing care for the indigent and working uninsured. PMID- 14556301 TI - New research highlights effects of Medigap reform. PMID- 14556302 TI - Enoxaparin in percutaneous interventions: when to monitor? A case series. PMID- 14556303 TI - Genetic testing: a family affair. PMID- 14556304 TI - Of genes and covenants. PMID- 14556305 TI - Validation of phage T7 biological dosimeter by quantitative polymerase chain reaction using short and long segments of phage T7 DNA. AB - Phage T7 can be used as a biological dosimeter; its reading, the biologically effective dose (BED), is proportional to the inactivation rate |ln (n/n0)|. For the measurement of DNA damage in phage T7 dosimeter, a quantitative polymerase chain reaction (QPCR) methodology has been developed using 555 and 3826 bp fragments of phage T7 DNA. Both optimized reactions are so robust that an equally good amplification was obtained when intact phage T7 was used in the reaction mixture. In the biologically relevant dose range a good correlation was obtained between the BED of the phage T7 dosimeter and the amount of ultraviolet (UV) photoproducts determined by QPCR with both fragments under the effect of five various UV sources. A significant decrease in the yield of photoproducts was detected by QPCR in isolated T7 DNA and in heated phage compared with intraphage DNA with all irradiation sources. Because the yield of photoproducts was the same in B, C and A conformational states of T7 DNA, a possible explanation for modulation of photoproduct frequency in intraphage T7 DNA is that the presence of bound phage proteins induces an alteration in DNA structure that can result in increased induction of photoproducts. PMID- 14556306 TI - Measurement of solar UVB variations by polysulphone film. AB - Sheets of polysulphone film have been extensively used as detectors to monitor solar UVB radiation. The advantages of polysulphone detectors are that they are small in size, they have good thermal stability and they are sensitive to UVB radiation. The principal disadvantage of polysulphone detectors is that their spectral sensitivity includes part of the short-wavelength UVA. In this study, we investigate the spectral sensitivity of the polysulphone detector with a series of monochromatic (+/- 2 nm) excitations. We then compare the polysulphone effective solar radiation with the erythemally effective solar radiation by comparing solar UVB data obtained with polysulphone films with those obtained with a spectroradiometer. From polysulphone data on the seasonal variation of solar UVB radiation, we estimate the corresponding fluctuations of the absorption of the ozone layer. We show that the spectral sensitivity of the polysulphone film is closer to the erythema action spectrum than that indicated by earlier data and that polysulphone detectors can be used to predict the erythema risk of solar UVB. Measurements on solar UVB with polysulphone films and with a spectroradiometer were found to be strongly correlated (R2 > 0.95). Finally, polysulphone-based measurements provide a good measure of the fluctuations of the stratospheric ozone layer. PMID- 14556307 TI - UV damage and photoreactivation: timing and age are everything. AB - Aquatic organisms, ranging from bacteria to fish, living in clear lakes are presently receiving damaging levels of UV radiation. Photoreactivation is a light dependent mechanism by which some organisms deal with DNA damage caused by UV radiation. Yet, photoreactivation is a mechanism that confounds long-term predictive modeling of UV effects on the survival of these organisms. Here we show that a short-lived rotifer species, Asplanchna girodi, previously thought to have little to no photoreactivation, does indeed have a significant amount of it. The ability to undergo photoreactivation in A. girodi is dependent on age and becomes apparent only after several days of observation after UV exposure. PMID- 14556308 TI - Viability of the antigen determines whether DNA or urocanic acid act as initiator molecules for UV-induced suppression of delayed-type hypersensitivity. AB - UV radiation suppresses the immune response, and UV-induced immune suppression contributes to UV-induced photocarcinogenesis. For UV-induced immune suppression to occur, electromagnetic energy (i.e. UV radiation) must be converted to a biological signal. Two photoreceptors have been identified in the skin that serves this purpose, epidermal DNA and trans-urocanic acid (UCA). Although compelling evidence exists to support a role for each pathway (UV-induced DNA damage or photoisomerization of UCA) in UV-induced immune suppression, it is not clear what determines which photoreceptor pathway is activated. To address this question, we injected UV-irradiated mice with a monoclonal antibody with specificity for cis-UCA or applied liposomes containing DNA repair enzymes to the skin of UV-irradiated mice. The effect that each had on UV-induced suppression of delayed-type hypersensitivity was measured. We asked whether the light source used (FS-40 sunlamps vs solar-simulated UV radiation) altered whichever pathway of immune suppression was activated. Different doses of UV radiation and the viability of the antigen were also considered. Neither the dose of UV nor the light source had any influence on determining which pathway was activated. Rather, we found that the viability of the antigen was the critical determinant. When live antigens were used, UV-induced immune suppression was blocked with monoclonal anti-cis-UCA but not with T4 endonuclease V-containing liposomes. The reverse was observed when formalin-fixed or killed antigens were used. Our findings indicate that antigen viability dictates which photoreceptor pathway predominates after UV exposure. PMID- 14556309 TI - Immunological aspects of photodynamic therapy of liver tumors in a rat model for colorectal cancer. AB - We have investigated tumor immunological effects of photodynamic therapy (PDT) of liver metastases. Livers of Wag/Rij rats were inoculated with three tumors of a syngeneic rat colon carcinoma cell line, CC531. One tumor in each rat was illuminated, with or without previous administration of the photosensitizer metatetrahydroxyphenylchlorin (mTHPC). PDT was effective in causing necrosis of tumors, but it did not affect the growth rate of nearby, nonilluminated tumors in the liver. Immunological staining of tumors showed natural killer (NK) cells to be significantly lower in PDT-treated tumors than in control tumors (P < 0.05). T cells in PDT-treated tumors and in their margins were lower than in tumors that received only sensitizer or only illumination (P = 0.015) at day 2 after treatment but reappeared at the tumor margins from day 7 after treatment. For macrophages, a similar pattern was found. NK cells, T cells or macrophages in nonilluminated tumors in mTHPC-treated rats did not increase significantly when compared with tumors in rats without mTHPC treatment. These findings indicated that no antitumor effect of a systemic immune response was present, as measured by the effect of PDT on growth of distant tumors and the number of T lymphocytes, NK cells and macrophages in these tumors. PMID- 14556310 TI - Ca(2+)-dependent and caspase-3-independent apoptosis caused by damage in Golgi apparatus due to 2,4,5,7-tetrabromorhodamine 123 bromide-induced photodynamic effects. AB - To clarify the role of the Golgi apparatus in photodynamic therapy-induced apoptosis, its signaling pathway was studied after photodynamic treatment of human cervix carcinoma cell line HeLa, in which a photosensitizer, 2,4,5,7 tetrabromorhodamine 123 bromide (TBR), was incorporated into the Golgi apparatus. Laser scanning microscopic analysis of TBR-loaded HeLa cells confirmed that TBR was exclusively located in the Golgi apparatus. HeLa cells incubated with TBR for 1 h were then exposed to visible light using an Xe lamp. Light of wavelength below 670 nm was eliminated with a filter. Morphological observation of nuclei stained with Hoechst 33342 revealed that apoptosis of cells was induced by exposure to light. Electron spin resonance spectrometry showed that light-exposed TBR produced both singlet oxygen (1O2) and superoxide anion (O2-). Apoptosis induction by TBR was inhibited by pyrrolidine dithiocarbamate, an O2- scavenger, but not by NaN3, a quencher of 1O2. Furthermore, TBR-induced apoptosis was inhibited by aurintricarboxylic acid and ZnCl2, which are known as inhibitors of deoxyribonuclease (DNase) gamma, and (acetoxymethyl)-1,2-bis(o-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid, a chelator of Ca2+, but not by acetyl Asp-Glu Val-Asp-aldehyde, an inhibitor of caspase-3. These results suggested that O2- was responsible for TBR-induced apoptosis, and Ca(2+)-dependent and caspase-3 independent nuclease such as DNase gamma played an important role in apoptotic signaling triggered by Golgi dysfunction. PMID- 14556311 TI - Photooxidations initiated or sensitized by biological molecules: singlet oxygen versus radical peroxidation in micelles and human blood plasma. AB - Biomolecules common in blood plasma, including 2-methyl-1,4-naphthoquinone (vitamin K-0, 2), 2,3-dimethoxy-5-methyl-1,4-benzoquinone (ubiquinone-0, 3), bilirubin, 4, and urocanic acid, 5, were used as photoactivators for the photooxidation of methyl linoleate (ML) in 0.50 M sodium dodecyl sulfate micelles to mimic a bioenvironment. UV irradiation of 2 in this system initiated H-atom abstraction from ML (Type-I mechanism). The evidence includes kinetics of oxygen uptake, inhibition of oxidation by an antioxidant ((R)-(+)-6-hydroxy-2,5,7,8 tetramethylchroman-2-carboxylic acid [Trolox], 7) and the analysis of four geometric hydroperoxides formed (cis, trans to trans, trans ratio of 0.5). In contrast, irradiation with a singlet-oxygen sensitizer, 3,5-di-t-butyl-1,2 benzoquinone, 1, formed six isomers by a Type-II mechanism, yielding a cis, trans to trans, trans isomer ratio of 6. Peroxidation activated by 3 or 4 with visible light occurred by a singlet-oxygen pathway (Type-II mechanism), as shown by kinetics of oxygen uptake and the effect of quenchers. In contrast, peroxidation in the presence of 5 in this system initiated H-atom abstraction from ML as shown by oxygen uptake and inhibition by Trolox. A comparison of thermal free-radical peroxidation with direct photooxidation of human blood plasma samples showed important differences. Blood plasma resisted thermal peroxidation because of natural antioxidants or on the addition of Trolox. In contrast, direct photooxidation involved singlet oxygen, according to the effect of quenchers and the lack of inhibition by antioxidants. PMID- 14556312 TI - Hesperetin glucuronide, a photoprotective agent arising from flavonoid metabolism in human skin fibroblasts. AB - There is considerable interest in the biological properties of flavonoids in terms of their antioxidant and cytoprotective actions. The interaction of the flavanone hesperetin with human skin fibroblasts (FEK4) has revealed the potential for metabolism to hesperetin glucuronide and its subsequent extrusion. As a consequence of this observation, the effectiveness of hesperetin glucuronides, in comparison with that of the aglycone form, in protecting against UV-A radiation has been investigated. The results indicate that hesperetin glucuronides, but not hesperetin, protect against UV-A-induced necrotic cell death. PMID- 14556313 TI - Chain-breaking antioxidant and cytoprotective action of nitric oxide on photodynamically stressed tumor cells. AB - Nitric oxide (.NO) has a multitude of physiological roles, including the ability to protect cells against oxidant-induced killing, e.g. by inhibiting caspase mediated apoptosis or by intercepting damaging free radicals derived from membrane lipids. The purpose of this study was to test the hypothesis that low flux .NO acting in the latter fashion can enhance tumor-cell resistance to photodynamic killing, specifically that sensitized by 5-aminolevulinic acid (ALA) derived protoporphyrin IX (PpIX). Preliminary model experiments with iron ascorbate-treated, PpIX-sensitized liposomes showed that spermine NONOate (SPER/NO)-derived .NO had no effect on photoinduced accumulation of primary singlet oxygen adducts, e.g. the cholesterol hydroperoxide 5 alpha-OOH, but dose dependently inhibited the buildup of free radical-generated oxidation products arising from one-electron turnover of primary peroxides. In subsequent studies, breast tumor COH-BR1 cells in serum-free medium were treated with 1 mM ALA for 15 min and then without ALA for 3.75 h, allowing biogenerated PpIX to diffuse to extramitochondrial sites, including plasma membrane. Cells were irradiated in the absence or presence of SPER/NO and compared for peroxidative damage and Hoechst assessed viability after 5 h in the dark. Iron-stimulated necrotic photo-killing and accumulation of chain lipid peroxidation products were observed, and this was inhibited strongly by SPER/NO, but not by decomposed SPER/NO, confirming that .NO was the active agent. When introduced after irradiation, .NO became progressively less inhibitory, consistent with ongoing but waning free-radical activity. These findings provide new insights into the possible role of .NO in tumor resistance to ALA-photodynamic therapy and other photo-dynamic treatments. PMID- 14556314 TI - Monitoring in situ dosimetry and protoporphyrin IX fluorescence photobleaching in the normal rat esophagus during 5-aminolevulinic acid photodynamic therapy. AB - Experimental therapies for Barrett's esophagus, such as 5-aminolevulinic acid (ALA)-based photodynamic therapy (PDT), aim to ablate the premalignant Barrett's epithelium. However, the reproducibility of the effects should be improved to optimize treatment. Accurate irradiation with light of a proper wavelength (633 nm), fluence and fluence rate has shown to be critical for successful ALA-PDT. Here, we have used in situ light dosimetry to adjust the fluence rate measured within the esophagus for individual animals and monitored protoporphyrin IX (PpIX) fluorescence photobleaching simultaneously. Rats were administered 200 mg kg-1 ALA (n = 14) or served as control (n = 7). Animals were irradiated with an in situ measured fluence rate of 75 mW cm-2 and a fluence of 54 J cm-2. However, this more accurate method of light dosimetry did not decrease the variation in tissue response. Large differences were also observed in the dynamics of PpIX fluorescence photobleaching in animals that received the same measured illumination parameters. We found that higher PpIX fluorescence photobleaching rates corresponded with more epithelial damage, whereas lower rates corresponded with no response. A two-phased decay in PpIX fluorescence could be identified in the response group, with a rapid initial phase followed by a slower rate of photobleaching. Non-responders did not show the rapid initial decay and had a significantly lower rate of photobleaching during the second phase of the decay (P = 0.012). PMID- 14556315 TI - Potentiation of photodynamic therapy with hypericin by mitomycin C in the radiation-induced fibrosarcoma-1 mouse tumor model. AB - Hypericin, a polycyclic quinone obtained from plants of the genus Hypericum, has been shown to be a promising photosensitizer. We investigated the combination of hypericin-photodynamic therapy (PDT) and a bioreductive drug mitomycin C (MMC) in the present study. The radiation-induced fibrosarcoma-1 tumors were exposed to laser light (120 J/cm2 at 595 nm) 24 h after an intravenous injection of hypericin (1 mg/kg). Hypericin-PDT alone significantly decreased tumor perfusion and oxygen tension as demonstrated by India ink staining technique and OxyLite pO2 measurement, respectively. The in vivo-in vitro cell-survival assay revealed about 60% direct tumor cell killing immediately after PDT. No significant delayed tumor cell death was observed after PDT, which suggests that vascular damage does not contribute significantly to the overall tumor cell death. Injection of a 2.5 mg/kg dose of MMC 20 min before light application significantly decreased tumor cell survival and delayed tumor growth compared with PDT or MMC alone. No greater skin reaction was observed after the combination of MMC and PDT than after PDT alone. Our study demonstrates that combining hypericin-PDT with MMC can be effective in enhancing tumor response with little side effect. PMID- 14556316 TI - Photochemical internalization enhances the cytotoxic effect of the protein toxin gelonin and transgene expression in sarcoma cells. AB - Further advantages in the treatment of soft-tissue sarcomas will only be achieved by tailoring the adjuvant therapy after surgery. The photochemically directed release of macro-molecules from endosomes and lysosomes into the cytosol is a novel technology, named photochemical internalization (PCI), that has been evaluated for treatment of sarcoma cells in vitro. Two human synovial sarcoma cell lines (SW 982 and CME-1) were treated with the photosensitizer meso tetraphenylporphine with two sulfonate groups on adjacent phenyl rings (TPPS2a) and a plasmid encoding enhanced green fluorescent protein (EGFP) complexed to poly-L-lysine to investigate the influence of PCI on gene transfer and with 5 micrograms/mL gelonin to investigate PCI of a Type-I ribosome-inactivating protein toxin. In addition, both cell lines were transduced with an Adenovirus serotype 5 encoding the Escherichia coli lacZ gene (AdHCMV-lacZ, expressing beta galactosidase) and treated with TPPS2a and light to evaluate the effect of PCI on the transduction rate. Photochemically induced transfection with the reporter gene EGFP in CME-1 cells increased from 0% of cells at no light to 40% of the cells after 60 s of light exposure. In contrast, the SW 982 cells showed no enhanced expression of the gene. The fraction of virally transduced cells was about doubled in both cell lines by means of PCI, although the transduction was more efficient in the CME-1 cells. Both cell lines became up to four-fold more sensitive to light when combining photochemical treatment with gelonin incubation. Our experiments showed that PCI induced the endocytic escape of therapeutic substances in cells derived from human soft-tissue sarcomas. PMID- 14556317 TI - Initial characterization of the primary photochemistry of AppA, a blue-light using flavin adenine dinucleotide-domain containing transcriptional antirepressor protein from Rhodobacter sphaeroides: a key role for reversible intramolecular proton transfer from the flavin adenine dinucleotide chromophore to a conserved tyrosine? AB - The flavin adenine dinucleotide (FAD)-containing photoreceptor protein AppA (in which the FAD is bound to a novel so-called BLUF domain) from the purple nonsulfur bacterium Rhodobacter sphaeroides was previously shown to be photoactive by the formation of a slightly redshifted long-lived intermediate that is thought to be the signaling state. In this study, we provide further characterization of the primary photochemistry of this photoreceptor protein using UV-Vis and Fourier-transform infrared spectroscopy, pH measurements and site-directed mutagenesis. Available evidence indicates that the FAD chromophore of AppA may be protonated in the receptor state, and that it becomes exposed to solvent in the signaling state. Furthermore, experimental data lead to the suggestion that intramolecular proton transfer (that may involve [anionic] Tyr 17) forms the basis for the stabilization of the signaling state. PMID- 14556318 TI - A photochemical study of (E,E,E,E)-2-[9-(2-hydroxyethyl)imino-3,7-dimethyl 1,3,5,7-decatrien-1-yl]- 1,3,3-trimethylcyclohexene, a derivative of all-trans retinal and ethanolamine. AB - A derivative of all-trans-retinal (RAL) and ethanolamine, A2-E, is the main fluorescent component of human retinal lipofuscin. The accumulation of lipofuscin has been correlated with exposure to ambient radiation and loss of photoreceptors. A possible precursor to A2-E is the imine formed from RAL and ethanolamine. This compound, (E,E,E,E)-2-[9-(2-hydroxyethyl)imino-3,7-dimethyl 1,3,5,7- decatrien-1-yl]-1,3,3-trimethylcyclohexene (HIDD), has been synthesized and structurally characterized. The photophysical and photochemical properties of HIDD and its protonated form, HIDD-H+, have been investigated using steady-state and time-resolved methods. Both HIDD and HIDD-H+ are weakly fluorescent, and the fluorescence lifetime and quantum yield for HIDD are ca 0.6 ns and 4.0 +/- 0.5 x 10(-4), respectively. HIDD forms a triplet excited state on direct excitation that decays with kd = 3.4 x 10(4) s-1. The extinction coefficient and quantum yield of intersystem crossing for the HIDD triplet are measured as 7.6 +/- 1.3 x 10(4) M-1 cm-1 and 0.055 +/- 0.006, respectively. The triplet excited state of HIDD-H+ can be sensitized by triplet energy transfer and has a decay rate constant of 1.6 x 10(4) s-1. The lifetime of the HIDD triplet excited state is observed to decrease with increasing concentration of the well-known electron or hydrogen atom donors, 2,3,5,6-tetramethyl-1,4-phenylenediamine and 2,3,5 trimethylhydroquinone, and the bimolecular rate constants for these reactions are approximately 5.4 x 10(6) M-1 s-1 and 1.7 x 10(8) M-1 s-1, respectively. These types of reactions may model photooxidative mechanisms of damage in the retina. PMID- 14556319 TI - Sequential effects of ultraviolet radiation on the histomorphology, cell density and antioxidative status of the lens epithelium--an in vivo study. AB - In vivo progressive effects of UV irradiation on the lens epithelium were studied using various histomorphological and biochemical parameters. Fifteen day old rat pups were exposed to 600 mW/m2 of radiation, including UV-A and UV-B, 12 h daily for 90, 120, 150 and 180 days. Biochemical parameters such as protein-bound and non-protein-bound sulfhydryl groups in both soluble and insoluble fractions and enzymes, which play an important role in combating the oxidative stress, were studied. Decreased cell density of lens epithelial cells (LEC) was observed in all three zones along with the decrease in the levels of soluble sulfhydryls (S SH), glutathione reductase (GR), superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT). On the other hand, an increase in insoluble sulfhydryls was observed. Because of the decrease in S-SH and GR activities, the LEC became vulnerable to oxidative stress. Decreased activities of SOD, GPx and CAT suggest elevated oxidative stress. This effect of UV radiation may lead to cell death that may be responsible for the observed decrease in the cell density in all three zones of the lens epithelium. PMID- 14556320 TI - Maternal health care at the crossroads. PMID- 14556321 TI - Pre-operative investigations: yield and conformity to national guidelines. AB - Routine ordering of pre-operative investigations yields a low true positive rate and is not cost effective. In this study, case notes of 251 adults who underwent elective surgery were reviewed. Pre-operative investigations were classified as 'indicated' or 'not indicated', based on the national guidelines. Only 56% of all tests done were indicated. The overall rates of expected and unexpected abnormal values from pre-operative blood investigations were 51.1% and 34.4% respectively. This study found that selective testing based on guidelines was beneficial. However, the results also suggest that the local guidelines need to be reviewed. PMID- 14556322 TI - Pre and post treatment mucociliary function in allergic rhinitis in three different treatment modalities. AB - Allergic rhinitis causes an impairment of the mucociliary function in the nose. It is hoped that treatment of perennial allergic rhinitis would be able to revert mucociliary function to normal. This study aims to compare pre and post treatment mucociliary transport time in 3 different treatment modalities. Ninety-two newly diagnosed patients with allergic rhinitis were randomised into 3 groups and started on different treatment regimes. At the end of 8 weeks, the group treated with only intranasal beclomethasone showed some, though not significant, improvement in the mucociliary function. There were no changes in the mucociliary function in the other two groups treated with beclomethasone and loratidine or loratidine alone. PMID- 14556323 TI - Urethritis in men at the Genito-Urinary Medicine Clinic Kuala Lumpur Hospital. AB - The clinical features and aetiology of 100 consecutive symptomatic heterosexual male patients with urethritis were studied from March 1994 to August 1994 in the Genito-Urinary Medicine (GUM) Clinic, Kuala Lumpur Hospital. Gonococcal urethritis (GU) was found to be more common (53%) than non-gonococcal urethritis(47%). All patients with GU confirmed microbiologically had clinically evident urethral discharge. Almost half (41%) of the patients with GU developed post-gonococcal urethritis (PGU). The most common organism isolated in PGU was Ureaplasma urealyticum (37%) whilst only 4% had both Chlamydia trachomatis and Ureaplasma urealyticum. Of the 47% of patients with non-gonococcal urethritis (NGU), 50% had no microorganism isolated, 32% had Ureaplasma urealyticum, 7% Chlamydia trachomatis and 11% both Chlamydia trachomatis and Ureaplasma urealyticum. PMID- 14556324 TI - Pregnancy in patients with renal transplants in Malaysia. AB - There were 72 pregnancies in 46 renal transplants (RTs) between 1984 and 2001, 89% from living donors, 11% cadaveric. Mean age at RT was 26.9 +/- 4.3 years and at pregnancy 30.7 +/- 4.7 years. Mean time to pregnancy after RT was 4.5 +/- 3.1 years. 54% were unplanned. 45 (63%) resulted in surviving infants, 37% delivered by Caesarean section. 35% were premature. Mean birth weight was 2.38 +/- 0.57 kg. 64% were on cyclosporine. No patient had an acute rejection during pregnancy; 38% had pre-existing hypertension. Complications include urinary infection (13%), proteinuria (15%) and preeclampsia (15%). Mean serum creatinine before pregnancy was 112.7 +/- 32.6 umol/l, 1 year post-pregnancy it was 119.4 +/- 38.7. The mean time of follow up of mothers is 4.9 +/- 3.5 years. 10 year graft survival was 83% and patient survival 94%. PMID- 14556325 TI - Did we do it right?--an evaluation of the colour coding system for antenatal care in Malaysia. AB - Identification of pregnancies that are at greater than average risk is a fundamental component of antenatal care. The objective of this study was to assess the level of appropriate management and outcomes among mothers with hypertensive disorders of pregnancy, postdates and anemia in pregnancy, and to determine whether the colour coding system had any effect on the maternal mortality ratios. A retrospective follow-through study confined to users of government health services in Peninsular Malaysia was carried out in 1997. The study areas were stratified according to their high or low maternal mortality ratios. The study randomly sampled 1112 mothers out of 8388 mothers with the three common obstetric problems in the selected study districts. The study showed that the prevalence of anemia, hypertensive disorders in pregnancy and postmaturity among mothers with these conditions were according to known international standards. There was no significant difference in the colour coding practices between the high and low maternal mortality areas. Inappropriate referrals were surprisingly lower in the areas with high maternal mortality. Inappropriate care by diagnosis and by assigned colour code were significantly higher in the areas with high maternal mortality. The assigned colour code was accurate in only 56.1% of cases in the low maternal mortality areas and in 55.8% of the cases in the high maternal mortality areas and these two areas did not differ significantly in their accurate assignment of the colour codes. The colour coding system, as it exists now should be reviewed. Instead, a substantially revised system that takes cognisance of evidence in the scientific literature should be used to devise a more effective system that can be used by health care personnel involved in antenatal care to ensure appropriate level of care and referrals. PMID- 14556326 TI - A community based study on the prevalence and factors affecting smoking in Kampong Jenderam Hilir, Sepang, Selangor. AB - OBJECTIVES: i) To determine the prevalence of smoking among the community aged 15 and above in Kg. Jenderam Hilir in Sepang District, Selangor state. ii) To determine the relationship between smoking and age, sex, household income and education. iii) To determine the parental and peer influence on smoking in Kg Jenderam Hilir. MATERIALS AND METHODS: A study was conducted in Kg. Jenderam Hilir, Sepang. All residents aged 15 years and above from all the 381 households of Kg. Jenderam Hilir were included in this study. A standardized pre-tested structured questionnaire was used in this study. RESULTS: Out of 894 respondents interviewed 69% were males compared to 3.9% of the females (overall 33.6%) reported to have smoked at least once in his/her lifetime. The difference was statistically significant. The mean duration of smoking was 17.7 years. The age at which smoking was first attempted ranged from 4 years to 51 years with a mean of 18.7 years. The mean initiation age for the males was significantly lower as compared to the mean initiation age for the females. The prevalence of current smokers was significantly higher amongst males (52.8%) as compared to the females (1.4%). The prevalence of smoking among current smokers increased with age (p < 0.01). There was no significant difference in the prevalence of smoking by household income and educational level. The mean number of cigarettes smoked by males was significantly higher than the females (p < 0.05). The mean duration of smoking among current smokers was 20 years. There was a significant difference in the level of addiction to smoking and the duration of smoking (p < 0.05). The high level of addiction increased with the duration of smoking (p < 0.05). Sixty five percent of the current smokers reported to have tried to quit smoking while 26% of ever smokers had reported to quit smoking and remained nonsmokers. The quit ratio was calculated to be 26.0%. The quit ratio was 52.6% among those with college or university education. There was a significant difference in the quit ratio by education level (p < 0.01). There was no significant difference in smokers whose parents were smokers as compared to nonsmokers whose parents were smokers. However, there was a significant association between smoking and peer influence (p < 0.01). PMID- 14556327 TI - Medial opening wedge high tibial osteotomy for osteoarthritis of knee: long-term results in 50 knees. AB - To retrospectively study the long term results of high tibial valgus osteotomy in management of primary medial compartment osteoarthritis, with special reference to patient satisfaction and functional assessment, we conducted this study in radiologically established 50 knees of patients with primary osteoarthrosis of knee. In these patients with medial compartment disease (varus knee), medial open wedge osteotomy was performed using full thickness iliac crest grafts. These were called for follow-up after average 7.5 years and clinico-radiological assessment was done. Although many surgeons of the west do not favour this osteotomy, we found the procedure to be quite acceptable to our patients in whom the primary concerns of cost and squatting habits are well taken care of. The authors feel that this osteotomy is still relevant in the third world. Results do deteriorate with time but most patients consider the surgery satisfactory. PMID- 14556328 TI - The use of nerve conduction studies in determining the short-term outcome of Bell's palsy. AB - Bell's palsy is a common neurological problem causing considerable loss of self esteem among patients. A prospective observational study was conducted to determine the short-term outcome of Bell's palsy at 1 month and 2 months after the onset and the relationship between these outcomes with facial nerve degeneration. We also determined if gender, age, diabetes, systolic and diastolic blood pressure influence the severity of facial nerve degeneration and the clinical outcome at 2 months after the onset. After clinically grading the newly diagnosed unilateral Bell's palsy patients using the House-Brackmann facial nerve grading system, nerve conduction studies of the facial nerve were done to determine the severity of facial nerve degeneration. The recovery of the facial paralysis was clinically graded again at the end of 1 month and 2 months from the onset. A total of 37 patients were recruited. There was a strong positive correlation between facial nerve degeneration and the clinical outcome of Bell's palsy at 1 month (r = 0.794; p < 0.0005) and 2 months (r = 0.732; p < 0.0005) after the onset. There was no significant correlation between either the facial nerve degeneration or the clinical outcome at 2 months with the patients' age (p = 0.288 and p = 0.799 respectively), systolic blood pressure (p = 0.425 and p = 0.933 respectively) or diastolic blood pressure (p = 0.243 and p = 0.579 respectively). Neither the severity of facial nerve degeneration nor the clinical outcome at 2 months were significantly different between male and female patients (p = 0.460 and p = 0.725 respectively) or diabetic and non-diabetic patients (p = 0.655 and p = 0.655 respectively). PMID- 14556329 TI - World Health Organization Quality of Life Assessment: brief version in Bahasa Malaysia. AB - WHOQOL-100, a 100 items quality of life assessment by WHO is too lengthy to be applicable in researches where the quality of life is one of the many variables of interest. The abbreviated version with 26 items is more acceptable by subjects, especially those with illness. The generic and the abbreviated Malay version were given to subjects who were healthy and with illness. Results showed that the domain scores produced by WHOQOL-BREF correlate highly with that of WHOQOL-100. WHOQOL-BREF domain scores demonstrated good discriminant validity, construct validity, internal consistency and test-retest reliability. The study indicates that WHOQOL-BREF in its brevity offers a valid and reliable assessment of quality of life. PMID- 14556330 TI - Paediatric admissions at a tertiary hospital in Kuala Lumpur--a case for a short stay ward. AB - The profile of admissions staying less than 24 hours admitted to the paediatric wards of University Malaya Medical Center, Kuala Lumpur, over a period of six weeks was reviewed to ascertain the need of a short-stay ward. Ninety-three (22%) of the 428 admissions admitted during the study period were discharged within 24 hours, 56 (60%) were discharged within 12 hours. Major categories of admissions were: elective investigative procedures (43%), and emergency admissions (44%). Reasons for emergency admissions: infections 42%, minor trauma/cerebral concussion 25% and febrile/afebrile seizures 11%. Only 20% required percutaneous oximetry monitoring and 2% required observations more frequently than 2 hourly. There may be a case for a short stay ward in a big paediatric unit in Malaysia. PMID- 14556331 TI - A retrospective review comparing the use of Gonal-F and Metrodin-HP for in-vitro fertilisation (IVF). AB - All cycles of IVF with pituitary down-regulation (n = 57) done at the Damansara Fertility Centre in the year 2000 were studied. All the 57 patients had controlled ovarian hyperstimulation, either using Metrodin HP (n = 27) or Gonal-F (n = 30). Of these, 53 patients reached oocyte pick-up, 26 patients in Metrodin HP group and 27 patients in Gonal-F group. Gonal-F resulted in a higher clinical pregnancy rate of 66.6% compared to Metrodin HP 38.5% (p < 0.05). The live birth rate tends to be higher in Gonal-F group (40.7%) compared to Metrodin HP (30.8%), (p > 0.05). PMID- 14556332 TI - Congenital diaphragmatic hernia in a post-partum woman. AB - Congenital diaphragmatic hernia (CDH) is rare in adults. We report a 24 year old woman presenting with shortness of breath, chest pain and nausea after the birth of her first baby. Clinical examination, plain radiography and a CT scan revealed herniation of abdominal contents into her left chest. Via a midline laparotomy, the contents were reduced and the defect repaired, using a mesh. She remains symptom-free three years since her surgery and even after a second childbirth. A brief review of the literature reporting adult diaphragmatic hernia of congenital origin accompanies this case report. We conclude that symptomatic CDH in adults usually presents as an emergency with gastrointestinal and occasionally respiratory complications. Early diagnosis and repair is essential to avoid subsequent morbidity and mortality. PMID- 14556333 TI - Leukaemia cutis presenting as leonine facies. AB - Acute leukaemia may rarely present as diffuse papules, nodules and plaques forming leonine facies. Leukaemia cutis generally carries a poor prognosis, and responds less well to chemotherapy. We described a case of acute myelomonocytic leukaemia presenting as leonine facies as a result of extensive cutaneous infiltration. The patient did not achieve haematological remission following standard induction chemotherapy and succumbed 6 weeks after the diagnosis was made. PMID- 14556334 TI - Spinal cord injury without radiological abnormality (SCIWORA). AB - SCIWORA or Spinal Cord Injury Without Radiological Abnormality; is a pre-MRI term that includes injuries to the spinal cord in the absence of radiological (plain radiographs, tomographs and CT scans) evidence of injury to the spinal column or cord. It occurs in skeletally immature spines because of the inherent plasticity of the bony structures in this age group. The prognosis is dependent on the extent of cord damage, and the role of active management is limited. A high index of suspicion is needed to establish a diagnosis. This diagnostic accuracy can be improved with the free availability of MRI scanning for the spines. We describe three cases of SCIWORA with a minimum follow-up of three years and a review of current literature. PMID- 14556335 TI - Giant cell arteritis with panocular involvement in an Indian male. AB - A case of giant cell arteritis with systemic and panocular involvement is reported here. This elderly Indian male presented with symptoms of unilateral temporal headache and intermittent jaw claudication for a month followed by diplopia and blurring of vision and later loss of vision in the right eye. The right eye showed some limitation of ocular movements, presence of relative afferent pupil defect, anterior segment ischaemic changes and anterior ischaemic optic neuropathy. Visual evoked potential showed an absent P1 wave while the left eye with normal 6/6 vision sowed a prolonged P1 wave. Fundus fluoresceine angiography showed delay in choroidal perfusion. His erythrocyte sedimentation rate (ESR) was 120 mm/hr and he was started on oral prednisolone. Superficial temporal artery biopsy obtained one week after starting steroids was positive for giant cell arteritis. Steroids led to the resolution of optic disc swelling, disappearance of anterior segment signs, full recovery of right ocular movements and no further deterioration of the fellow eye. On steroids, he developed insomnia and progressive myopathy which resolved and is now symptom free at lower doses of steroids. PMID- 14556336 TI - Two-stage management of mega occipito-encephalocele. AB - We report three cases of large occipito-encephaloceles that were managed in the Neurosciences Unit, Hospital Universiti Sains Malaysia over the last 5 years. All patients had pre-operative MRI (magnetic resonance imaging) evolution and mapping of the sagittal sinus tract. The cerebrospinal fluid (CSF) of all three patients were initially diverted by means of ventricular shunt two weeks prior to shunt removal. The slow drainage of CSF prevented electrolyte and volume disturbances due to sudden decompression during their definitive occipital encephalocele operations. After 3 years follow-up, all these patients are progressing well. PMID- 14556337 TI - Metastatic disease of the proximal femur. AB - Since January 1999, ten patients had undergone surgical treatment for metastatic bony lesions of proximal femur at this centre. Seven of these patients were treated for complete pathological fractures, one for impending fracture and one for revision of internal fixation and loosening of hemiarthroplasty. Primary malignancies were located in breast in four cases, prostate in three and one in lung, thyroid and neurofibrosarcoma. Two patients had died within six months after surgery, four after 1 year while the remaining four were still alive. The mean duration of survival was eleven months. Nine patients had been ambulating pain free and there were no failure of reconstruction. PMID- 14556338 TI - Spontaneous rectus sheath haematoma in pregnancy. AB - Bleeding into the rectus sheath is an uncommon but a well-recognised condition that mimics several other diagnoses of acute abdomen. A wide range of etiology has been proposed in association with this condition. It is often self-limiting, but can lead to unnecessary laparotomy if the diagnosis is not recognised. PMID- 14556339 TI - Uterine artery bleeding and haemoperitoneum during the second trimester in pregnancy. AB - A case of spontaneous rupture of uterine artery in the second trimester of pregnancy is described. Haemorrhage from rupture of uterine artery during pregnancy was discovered at laparotomy. This was an unusual but serious complication of pregnancy. This condition is extremely rare and one must consider it in cases of incomprehensible abdominal pain with or without haemodynamic collapse. A review of the literature revealed only four similar cases so far. This pregnancy continued till 37 weeks pregnancy and had a spontaneous vaginal delivery. Immediate institution of effective resuscitative measures and early surgical intervention were essential to both foetal and maternal survival. PMID- 14556340 TI - Relapsed Hodgkin's disease presenting as a right knee swelling. AB - A 49 year-old Indian housewife was diagnosed with Hodgkin's disease in 1995. She was given combination chemotherapy comprising Chlorambucil, Vincristine, Procarbazine and Prednisolone. Unfortunately she defaulted after two courses of chemotherapy. One year later, she developed progressive right knee swelling and pain, associated with loss of appetite, loss of weight, intermittent fever, night sweats and pruritus. The right knee swelling measured 15 cm x 20 cm and was warm and tender. A plain radiograph of the right knee revealed osteolytic lesions at the distal end of the right femur and the proximal ends of the right tibia and fibula, associated with gross periosteal reaction and soft tissue swelling. Apart from left cervical lymphoadenopathy, examination of other systems was unremarkable. Pelvic bone marrow biopsy was inconclusive. An open biopsy of the lower end of the right femur was consistent with Hodgkin's disease. She was given salvage combination therapy comprising Chlorambucil, Vincristine, Procarbazine, Prednisolone, Doxorubicin, Bleomycin and Vinblastine. She tolerated the treatment well and responded with significant reduction in the swelling and pain of the right knee. Unfortunately, she again defaulted treatment after 2 courses of chemotherapy. This case illustrates an unusual presentation of Hodgkin's disease in relapse. PMID- 14556341 TI - An uncommon cause of duodenal obstruction. AB - This is a case report of an elderly male, who presented with duodenal obstruction caused by abdominal aortic aneurysm. An axillo-bifemoral shunting with an infra renal straight inlay graft together with a feeding jejunostomy was performed. This is rare presentation with only a few cases reported. PMID- 14556342 TI - Eagle's syndrome. AB - Eagle's syndrome is an uncommon condition resulting from an elongated styloid process, which causes cervico facial pain, tinnitus and otalgia. A 48-year-old female presented to the clinic with bilateral upper neck pain radiating to the ears with tinnitus for almost one-year duration. Examination of the oral cavity revealed atrophic tonsils and palpable bony projection deep in the tonsillar fossa. Plain lateral neck X-ray and CT scan confirmed the presence of bilateral elongated styloid processes, which were subsequently resected surgically through an oropharyngeal approach. The patient was asymptomatic at follow up at 2 years. PMID- 14556343 TI - Dengue antibodies in a suburban community in Malaysia. PMID- 14556344 TI - Palliative care for lung cancer patients in Penang Hospital. PMID- 14556346 TI - Medicine and the law. AB - When a doctor is required to go to court, he does so with some amount of trepidation. The degree of trepidation increases in direct proportion as to whether he is required to be a witness or a defendant. The practice of medicine on the other hand requires the patient to have full confidence and open out his secrets to the doctor. If you hold back vital information, the diagnosis may be entirely different to the disease that you have. Lawyers who enter hospitals may also do so with some trepidation, maybe even more so than doctors who enter courts, as their lives are at stake. There is a perception that medico-legal matters are on the rise. We may put forward a few reasons for this: 1. A better educated and increasingly assertive public with greater awareness of the medical and legal systems; 2. Rising expectations of medical results; 3. Commercialization of medical care with erosion of the doctor-patient trust relationship. This paper will discuss the reasons for and the ways to address medical errors as well as explore the reasons for defensive medicine. The argument is put forward that public education programs on the risks inherent in some of the new advances in treatment modalities and surgery and professional education programs on the need for obtaining the patient's informed consent to such treatment is needed. Public advocacy programs to demonstrate the problems in medicine and the delivery of health care resulting from strict cost containment limitations should be carried out. There is also the need to enhance the level and quality of medical education for all physicians, including improved clinical training experiences. Doctors' must manage their clinical affairs in a professional manner without being dictated to by the legal system. However, it would be wise to take note of the views expressed by learned counsel and judges in their courts. The middle road is always the best and we must never be extreme in our viewpoints. We must always remember the patient is why we are here and the patient must never suffer in the process while we formulate our responses to the medico-legal challenges that lie ahead. PMID- 14556345 TI - Social drug use in the parturient: implications for the management of obstetrical anaesthesia. AB - Maternal use of social drugs in pregnancy continues to increase--worldwide. Although a great deal has been learned regarding the implications of illicit drug abuse in pregnancy (cocaine, amphetamines, hallucinogens), the use of social drug in pregnancy has received far less attention. This article reviews the consequences of the social drug use in pregnancy including ethanol, tobacco and caffeine and offers recommendation for anaesthetic management of these potentially complicated pregnancies. PMID- 14556347 TI - The law and medicine--the right to healthcare and treatment. PMID- 14556348 TI - Unwanted pregnancy--medical and ethical dimensions. AB - Globally, abortion mortality accounts for approximately 13% of all maternal mortality. Unsafe abortion procedures, untrained abortion providers, restrictive abortion laws and high maternal mortality and morbidity from abortion tend to occur together. Unplanned and unwanted pregnancies constitute a serious public health responsibility. While fertility has declined by half in developing countries, the motivation to control and space births has risen faster than the rate of contraceptive use. Preventing maternal mortality and morbidity from abortion in countries where these remain high is a matter of good public health policy and medical practice, and constitutes an important part of safe motherhood initiatives. A range of positive steps has been taken to reduce deaths and morbidity from abortion in a growing number of countries over the past 15 years. Making abortion legal is an essential prerequisite in making it safe. In this respect, changing the law does matter and assertions to the contrary are ill conceived and unsupported in practice. Although, in many countries, trends towards safer abortion have often occurred prior to or in the absence of changes in the law, legal changes need to take place if safety is to be sustained for all women. Religious laws may also require attention when legal change is being contemplated. There are three main ways of approaching this problem: liberalizing the existing law within the penal or criminal code; partially or fully legalizing abortion through a positive law or a court ruling; and decriminalising abortion by taking it out of the law. Women's health groups and other advocates, parliamentarians and health professionals, can work together to support the right of women not to die from unsafe abortions and to ensure they receive treatment for complications. Committed doctors can make a difference by providing treatment for abortion complications, interpreting the law in a liberal way and providing safe services where these are legal as well as training providers in the safest techniques to reduce mortality and morbidity. Although law, policy and women's rights are central to this issue, making abortions safe is above all a public health responsibility of governments. Moreover, reducing maternal mortality by making abortions safe is also an important part of the international commitment made in Cairo in 1994 at the ICPD and reaffirmed at the Cairo meeting in 1999. PMID- 14556349 TI - The legal aspects of the termination of unwanted pregnancies and the risks faced by the medical doctor: a UK perspective. AB - Historical perspective of terminations of unwanted pregnancies in the UK. Moral and ethical considerations imposed by established church's teachings becoming increasingly in conflict with the wishes and expectations of a more secular society. Recognition that illegal abortion was, as a matter of fact available, at great risk to vulnerable girls and women. Eventually public demand and a radical and reforming government led to the current Statutory Framework. Statutory provisions: Offences against the Person Act 1861, Sections 58 and 59; Infant Life Preservation Act 1929 Section 1. Recognition of the limited flexibility allowed by the law in the original restrictive statutory framework. The direction to the jury in July 1938 by Macnaghten J in the case of R. v. Bourne [1939] 1 KB 687, where an eminent obstetrician was acquitted after carrying out an abortion on a young rape victim. Then the modern statutory provisions: Abortion Act 1967, amended by the Human Fertilisation and Embryology Act 1990. The statutory framework provides for healthcare professionals not to have to take part in terminations if they have a conscientious objection to doing so. While there are still fierce challenges from moral pressure groups when any changes in the detail of the law are proposed--such as reducing the maximum gestation period for a lawful termination--as a whole society seems to have accepted the current law. Issues affecting doctors who consider and provide terminations; current medico legal problems relating to wanted pregnancies that have been lost by reason of clinical negligence, and unwanted children that have been born by reason of clinical negligence. PMID- 14556350 TI - Social and religious dimensions of unwanted pregnancy: an Islamic perspective. AB - The concept of 'unwanted pregnancy' is a recent in human history and is associated with social stresses of modern life. The purposes of the law, maqasid al shari'at, and its principles, qawa'id a shari'at, focus on preventing 'unwanted pregnancy', protecting the rights of the fetus and infant, and mitigating the adverse effects of 'unwanted pregnancy' by social measures. 'Unwanted pregnancy' is associated with general social determinants (hedonistic life styles, sexual transgression, addiction to drugs, fear of poverty, and low female status) and specific antecedent causes (sexual crimes, egoistic greed, maternal/fetal disease, and gender discrimination). It is prevented by sexual hygiene, marriage, contraception, deterring sexual crimes, and raising the status of women. The adverse sequelae of 'unwanted pregnancy' (feticide, infanticide, or child abuse and neglect) can be prevented by defending the basic human right of the fetus and infant to life, promoting social institutions for child welfare (nuclear family, extended family, foster care, and open adoption). Closed adoption is forbidden by Law but care in a foster home is allowed and is encouraged if the nuclear and extended families are unwilling or are unable to care for children. Abortion at any stage of pregnancy is a crime against humanity. It is not a solution to the problem but is part of the problem. It will encourage more 'unwanted pregnancies'. PMID- 14556351 TI - The legal dimensions of medical negligence. PMID- 14556352 TI - Medical misadventures. AB - Historical perspective of terminations of unwanted pregnancies in the UK. Moral and ethical considerations imposed by established church's teachings becoming increasingly in conflict with the wishes and expectations of a more secular society. Recognition that illegal abortion was, as a matter of fact available, at great risk to vulnerable girls and women. Eventually public demand and a radical and reforming government led to the current Statutory Framework. Statutory provisions: Offences against the Person Act 1861, Sections 58 and 59; Infant Life Preservation Act 1929 Section 1. Recognition of the limited flexibility allowed by the law in the original restrictive statutory framework. The direction to the jury in July 1938 by Macnaghten J in the case of R. v. Bourne [1939] 1 KB 687, where an eminent obstetrician was acquitted after carrying out an abortion on a young rape victim. Then the modern statutory provisions: Abortion Act 1967, amended by the Human Fertilisation and Embryology Act 1990. The statutory framework provides for healthcare professionals not to have to take part in terminations if they have a conscientious objection to doing so. While there are still fierce challenges from moral pressure groups when any changes in the detail of the law are proposed--such as reducing the maximum gestation period for a lawful termination--as a whole society seems to have accepted the current law. Issues affecting doctors who consider and provide terminations; current medico legal problems relating to wanted pregnancies that have been lost by reason of clinical negligence, and unwanted children that have been born by reason of clinical negligence. PMID- 14556353 TI - Medical care for people under detention. AB - Human Rights traditionally refer to rights and freedom that are inherent to every human being. They are based on Human Rights Law and concern the respect for dignity and worth of a person. These rights are universal, inalienable, indivisible, inter-related and interdependent. Members of Societies are detained for varied reasons and are made up of different age groups and gender. The United Nations through its numerous agencies, associated Conventions, Treaties and Resolutions have laid down guidelines that govern the rights of those under detention. Article 5 of General Assembly Resolution 45/111 clearly stipulates that except for those limitations that are demonstrably necessitated by the fact of incarceration, all prisoners shall retain the human rights and fundamental freedom set out in the Universal Declaration of Human Rights. As such, the Medical and Health Care of People under Detention should not be any different from the other members of societies. The Right to Health and Medical Care is stipulated under various Articles contained in the UN Bill of Human Rights (UDHR, ICCPCR and ICESCR) as well as other Conventions, e.g. Convention against Torture (CAT), Convention on Rights of the Child (CRC) and Convention for the Extinction of all Forms of Discrimination against Women (CEDAW). The United Nations have also developed specific guidelines and instruments for Treatment of People under Detention. These include the General Assembly Resolution 45/111 December 1990 elucidating the Basic Principles for Treatment of Prisoners, ECOSOG resolution 663C and 2076 regarding the Standard Minimum Rules for the Treatment of Prisoners which covers rules pertaining to accommodation and Medical Services, General Assembly Resolution 37/194 on Principles of Medical Ethics relevant to the role of health personnel, particularly Physicians, in the Protection of Prisoners and Detainees against Torture and Other Cruel, Inhuman or Degrading Treatment or Punishment. PMID- 14556354 TI - The importance of effective communication in preventing litigation. AB - Good communication in all aspects of medical practice is essential. Effective communication with your patient not only enables you to take an accurate history but also helps the patient to understand their illness and no doubt assists the healing process. Communication with other health professionals allows the team approach to healthcare to succeed. It reduces the chances of a breakdown in continuity of care, builds relationships and understanding between different disciplines and specialties and helps professionals to learn from each other. In the medico-legal field poor communication is the underlying problem in the most of cases that MPS deals with. The majority of negligence cases are not related to the clinical quality of care but are triggered by inadequate communication. A breakdown in the doctor-patient often occurs before the incident that leads to a claim. It is as if the patient is just waiting for their moment to sue. In a busy clinic or GP surgery it is often easy to forget the human needs of the patient and concentrate on their medical needs. First impressions are vital not just with the doctor but also with other staff and even the clinic or hospital itself. During the consultation careful listening, giving sincere empathy early in the consultation and an expression of understanding of their concerns will go a long way to instill confidence in the patient and reduce the likelihood of a complaint should things go wrong. PMID- 14556355 TI - The role of hospital management in reducing medical errors. PMID- 14556356 TI - The patient's view. AB - The medical practitioner has always had to juggle several roles. First and foremost, the doctor is a healer, a provider of curative services. Second, he is an examiner, an assessor of the patient's health status. Third, he is a researcher, always trying to push the boundaries of medical knowledge. Fourth, he is a rationer of services, he decides how best to apportion the limited resources at his disposal. Traditionally, the patient-doctor relationship has been largely exclusive in nature and the doctor would quite comfortably slip in and out of these roles, his focus centred on his patient's interests. In this era of large corporate health care providers, multi-billion-biotechnology industry, mammoth pharmaceutical companies, medical insurance schemes and international trade instruments, it has become increasingly difficult for the doctor to juggle these four roles. He is constantly subjected to conflicting demands. Patients' interests do not always come first anymore and patients are beginning to realise this. They no longer trust the medical profession unreservedly. There has been steady erosion of the patient-doctor relationship most clearly evidenced by the rising tide of litigation against doctors. There needs to be a reappraisal of these roles that the doctor plays. The conflicts must be recognised and addressed. Patients need to be informed and their interests must be protected if the doctor-patient relationship is to be restored. Medical malpractice suits are on the increase. The tort system as it exists is failing both doctors and patients. The question we must ask is what are patients looking for when they sue doctors? Most of the time they need compensation for the injuries suffered. Sometimes they are looking for accountability, they want the doctor to be punished in some way. Sometimes they merely want to air their grievances and know that they are heard. The current system more often than not takes too long to compensate, the process is a gamble and doctors who are clearly negligent quietly settle and are rarely censured. We need to revamp the existing system to allow for speedy and equitable compensation; true accountability; and articulation and auditing of standards of practice. PMID- 14556357 TI - Therapeutic and reproductive cloning--implications and recommendations. AB - The announcement of Dolly's birth took the world by storm, mainly because what was thought impossible has become possible. Optimism that new approaches in agriculture and medicine abound, as much as fear and imagination leading to Frankenstein-like scenarios. Scientifically, cloning refers to replicating an animal with the same nuclear genetic material; whilst it may refer to embryos as the source material, the current storm refers to differentiated ("somatic") cells. Cloning technology is useful in the following areas: agriculture, to produce animals of superior or specific qualities; endangered animals, to increase genetic diversity through widening the gene pool; understanding fundamental questions in developmental embryology, through the use of laboratory animals; and in human therapy, to produce cells and possibly tissues for repair and regeneration. In the first 2 instances, it is reproductive cloning. In the last instance, it is therapeutic cloning, as no individuals are "produced". Human reproductive cloning is not allowed by all governments that have deliberated on it, and therapeutic cloning is allowed by some under certain circumstances. As therapeutic cloning has great potential in cures of many diseases, it should be allowed but with safeguards to prevent abuse and reproductive cloning in the human. PMID- 14556358 TI - Ethical issues in stem cell research. AB - Like most cutting edge medical technology, human stem cell research raises a number of difficult and important ethical issues and concerns, requiring potential benefits to be balanced against the need to protect the rights and welfare of citizens. Much of the debate involves research using embryonic stem (ES) cells, which in turns revolves around the moral status of the human embryo, and the level of respect and protection that should be accorded. This is an especially sensitive issue in pluralistic societies where different, if not conflicting, cultural and religious perspectives exist. Another contentious issue as far as the derivation of ES cells is concerned is the intent involved in producing the embryos, specifically whether it is ethically permissible to allowing embryos to be made solely for the purpose of research. These and several other relevant ethical issues will be discussed, including a comparison of guidelines and positions adopted in different countries. PMID- 14556359 TI - Appropriate medical care for persons in detention. AB - The people who are in detention are screened by the Medical Officer of the Prison and if they are found to be unwell, these prisoners will be accommodated in the sickbay and medical treatment will be provided. If their sickness needs further investigations and management, they will be sent to the Government Hospital. If the prisoners are found to have infectious or contagious diseases, steps will be taken to prevent the spread of these diseases to other prisoners in the prison. Prisoners are given time to exercise to maintain good health and their clothing are regularly washed to make sure that they will not contract skin diseases, e.g. scabies, ringworm, etc. The Prison Department since 1989 has increasing numbers of HIV positive prisoners. The Department complies with this problem by sending staff for courses, lectures and seminars so that they will be able to handle these prisoners more efficiently in the prison. When these HIV/AIDS prisoners' condition turns bad, they are usually transferred to a Government Hospital. Another of the Prison Department's prominent medical problem among the prisoners is drug addiction. Staff trained with skill and techniques are counselors for the drug related prisoners. Realizing and in anticipation that the sickbays in the prisons are going to be full of HIV/AIDS prisoners and drug related prisoners, special attention will be given to more allocation to upgrade the sickbays in the prison. White attires will be provided to the sick prisoners in the sickbays so that they will look neat and clean. More doctors, medical assistants and nurses will be employed so that appropriate medical care or rather more appropriate medical care can be provided to the sick prisoners in the prisons. The Prison Department is in the process of privatizing medical care for prisoners in the prison and the Department is also trying to convert some prisons to be medical prisons so that adequate medical care can be given to the sick prisoners. PMID- 14556360 TI - Legal minimum standards of medical care for people in detention in Malaysia. PMID- 14556361 TI - Contemporary issues in risk management: "risk management for dentists". AB - The management of the clinician who generates complaints and claims on a regular basis, raises issues of professional responsibility and presents ethical challenges for the defence team, in addition to the immediate practical need for advice and representation. This short session examines some of these issues and suggests some ways that are already being used to address them. PMID- 14556362 TI - Providing protection for doctors. AB - Over the last decade the number of negligence cases brought against doctors, dentists and other healthcare professional has increased significantly in many countries around the world. In addition, the proliferation of the pathways of accountability in many countries means that doctors are feeling undervalued and vulnerable and are therefore seeking assistance and protection more than ever. The effect of a claim or complaint on a doctor can be devastating both personally and professionally. The relationship with the patient is damaged and an increase in defensive practice ultimately causes an increase in healthcare costs. Patient expectations have increased. Patients are more knowledgeable than they were five years ago and the demands on the professions are increasing daily. Unfortunately because of the long delays that are seen between an incident and a claim it is very difficult to predict what reserves are required in the future. We have seen a number of organisations pull out of the business over the last year and it highlights the importance of having an understanding of the needs of the professions and the ability to track trends in litigation in order to set appropriate subscription rates. Doctors and other healthcare professionals require more than just financial protection. They need support from their colleagues and assistance from experienced lawyers in order to help restore their confidence and ensure that they can continue to practice for the benefit of their patients in the future. PMID- 14556363 TI - Resolutions of the commonwealth medico-legal conference. PMID- 14556364 TI - Evaluating trihalomethane content in drinking water on the basis of common monitoring parameters: regression models. AB - The presence of trihalomethanes (THMs) in potable-water sources is an issue of great interest because of the negative impact THMs have on human health. The objective of this study was to correlate the presence of trihalomethanes with more routinely monitored parameters of water quality, in order to facilitate THM control. Water samples taken at various stages of treatment from a water treatment plant were analyzed for the presence of trihalomethanes with the Fujiwara method. The data collected from these determinations were compared with the values obtained for free-residual-chlorine and combined-residual-chlorine levels as well as standard physico-chemical and microbiological indicators such as chemical oxygen demand (by the KMnO4 method), total chlorophyll, conductivity, pH, alkalinity, turbidity, chlorides, sulfates, nitrates, nitrites, phosphates, ammonia, calcium, magnesium, heterotrophic bacteria count, Pseudomonas spp., total and fecal coliforms, and fecal streptococci. The data from these determinations were compiled, and statistical analysis was performed to determine which variables correlate best with the presence and quantity of trihalomethanes in the samples. Levels of THMs in water seem to correlate directly with levels of combined residual chlorine and nitrates, and inversely with the level of free residual chlorine. Statistical analysis with multiple linear regression was conducted to determine the best-fitting models. The models chosen incorporate between two and four independent variables and include chemical oxygen demand, nitrites, and ammonia. These indicators, which are commonly determined during the water treatment process, demonstrate the strongest correlation with the levels of trihalomethanes in water and offer great utility as an accessible method for THM detection and control. PMID- 14556365 TI - Aerobic residential onsite sewage systems: an evaluation of treated-effluent quality. AB - This retrospective cohort study used existing data to evaluate the quality of effluent from three of the most common types of onsite residential aerobic treatment sewage systems (Multi-Flo, Norweco, and Whitewater) installed in Kitsap County, Washington. Five-day biochemical oxygen demand (BOD5), total suspended solids (TSS), and fecal-coliform-bacteria parameters were used to determine performance. Although most (77 percent) of the systems were less than one year old at the time of sampling, over a third failed to meet NSF certification standards for BOD5 and TSS in effluent (< 30 milligrams per liter [mg/L]). Over two-thirds of systems failed to meet Washington State Board of Health Treatment Standard 2 criteria for BOD5 and TSS (< 10 mg/L). Furthermore, an average of 59 percent of the systems failed to meet state standards for fecal coliform (< 800 fecal coliform bacteria per 100 milliliters). PMID- 14556366 TI - NEHA's 2003 Annual Educational Conference and Exhibition-Reno, Nevada. PMID- 14556367 TI - Housing, inspection, and statute-of-limitations issues. PMID- 14556368 TI - Anthrax--CDC review. PMID- 14556369 TI - Bioterrorism early-warning system--algae? PMID- 14556370 TI - New emergency and disaster preparedness course work for physicians and other health care professionals. PMID- 14556371 TI - Adult day centers: everything you wanted to know but were afraid to ask! AB - Adult day services are a growing component of the wide spectrum of home and community-based care. However, up until now there has been only sketchy information about how many centers there are, what they do, whom they serve, and how they operate. This research brief presents findings from a new study by Partners in Caregiving: The Adult Day Services Program at the Wake Forest University School of Medicine, which provides the most comprehensive information to date about this important service. Understanding what centers do and whom they serve can be the start of a beautiful relationship. PMID- 14556372 TI - Setting the standard for community-based care & creating a comprehensive continuum of care. AB - Home health and adult day services providers are natural collaborators, as both institutions seek to allow chronically ill patients to receive care at home. Well formed partnerships between these two types of providers can result in better patient care that is more economical under the prospective payment system, and open up joint marketing opportunities. The authors review the benefits of home health-adult day services collaboration, as well as some considerations for home health agencies entering into an agreement with an adult day services provider. PMID- 14556374 TI - A step-by-step guide to home telehealth program planning. AB - Many home health providers have become familiar with the idea of home telehealth service, but planning its optimal delivery nevertheless presents a challenge. This article presents the first steps agencies need to take before the "televisiting" can begin. PMID- 14556373 TI - Financial success under PPS: how to beat the odds, Part II. AB - Whether you are a publicly held company or a small mom and pop agency, you have to maximize both efficiency and productivity while maintaining patient satisfaction and staff morale. In the new world of home care, this is a tall order. In the first installment of this two part series, which ran in the July 2002 issue of CARING the author reviewed the dimensions of financial success, issues of length of stay and episode of care, resource utilization planning, and reducing overhead. This second part covers overall productivity, clinical productivity, and management strategies to synergize financial success under the prospective payment system. PMID- 14556375 TI - Non-competition agreements: how to make them work for you. AB - In a time of staffing shortages and increased competition for shrinking reimbursement dollars, home care agencies need to protect themselves from unprincipled or unknowing employees that leave the agency to work for a competitor or for a patient, bringing with them other agency employees, patient lists, and other proprietary information. Well-drafted, attorney-reviewed, enforceable non-competition agreements can help mitigate some of these problems. PMID- 14556376 TI - A tribute to Representative Ben Cardin. PMID- 14556377 TI - Connecting with the cognitively impaired: dementia and Alzheimer's disease. AB - When confronted with their own or their loved one's cognitive decline, many people attribute it to "normal aging." Home care providers, due to their regular interaction with their clients, may be the first to identify dementia. As such, home care providers can play an important role by encouraging those involved to address the problem and develop a plan of care to help them to successfully navigate the challenges they face. Alzheimer's disease is one of the most common causes of dementia. This article identifies the warning signs of Alzheimer's disease and provides advice to home care providers on intervention and how to care for patients who are cognitively impaired. PMID- 14556378 TI - Power publicity: the ultimate referral source. PMID- 14556379 TI - Remembering Mother Teresa. PMID- 14556381 TI - A new way of analysing symmetrical and asymmetrical gaits in quadrupeds. AB - This new approach highlights for the first time that all the gaits, symmetrical as well as asymmetrical, correspond to the succession of sequences that start by the movements of the two fore limbs, followed by the movements of the two hind limbs. Inside those sequences, the gaits are identified by the time lag between the movements of the two pairs and by the time lag between the movements of the two feet inside each pair. This approach, by breaking the stride paradigm, gives a new framework to locomotion analysis. PMID- 14556380 TI - Identification of new partners of the epithelial sodium channel alpha subunit. AB - A fine regulation of the amiloride-sensitive Epithelial Sodium Channel (ENaC), made of alpha, beta and gamma subunits, is crucial for maintenance of Na+ balance and blood pressure. Both beta- and gamma-ENaC participate in negative regulation by interacting with Nedd4-2, an E3 ubiquitin-ligase. Disruption of this interaction results in increased ENaC activity (Liddle syndrome). By two-hybrid screenings, we identified new potential partners of alpha-ENaC: WWP1 (E3 ubiquitin-ligase protein), UBC9 and TSG101 (E2 ubiquitin/SUMO-conjugating enzymes) and confirmed these interactions in GST pull-down assays. All these partners are implicated in protein trafficking and could be involved in the regulation of ENaC activity. PMID- 14556382 TI - [Role of man in the history of vegetation of the Ellero valley (maritime Alps, Italy)]. AB - The pollen analysis of a sediment core from a peat bog (Rifugio Mondovi) at the mountain belt (1760 m) in the Ellero Valley (Italian Maritime Alps) shows the postglacial vegetation history. The sequence starts at 12,000 BP during a peak of pine pollen; this first phase shows a low representation of birch and the presence of Tilia. Younger Dryas is characterised by increasing percentages of Artemisia, showing the presence of deciduous Quercus, fir and beech. Elm appears at the beginning of the Holocene during the second pine peak (9800 BP). A 3000 year hiatus is present. Sedimentation resumes at 6000 BP in a Rhododendron fir wood. The present timberline at 1500 m, at the limit of the beech wood, is a result of the decline of the fir-wood at 2600 BP, which allowed an expansion of beech. During this period, there was a continual increase in Gramineae and deciduous oak and the first occurrences of evergreen oak are observed. The development of larch occurs at 1800 BP, together with walnut, chestnut, cereals and vine. PMID- 14556383 TI - Changes in antioxidant and lignifying enzyme activities in sunflower roots (Helianthus annuus L.) stressed with copper excess. AB - Treatment with 50 microM CuSO4 for five days caused significant decrease in dry matter production and protein level of ten-day-old sunflower seedling roots. An increase of lipoperoxidation product rate was also observed. The involvement of some enzyme activities in the sunflower root defence against Cu-induced oxidative stress was studied. Copper treatment induced several changes in antioxidant enzymes. SOD (superoxide dismutase, EC 1.15.1.1) activity was reduced but CAT (catalase, EC 1.11.1.6) and GPX (guaiacol peroxidase, EC 1.11.1.7) activities were significantly enhanced. The lignifying peroxidase activities, assayed using coniferyl alcohol and syringaldazine, were also stimulated. Analysis by native gel electrophoresis of syringaldazine peroxidase activity showed the stimulation of an isoform (A2) and the induction of another one (A1) under cupric stress conditions. On the other hand, the activity of PAL (phenylalanine ammonia lyase, EC 4.3.1.5), which plays an important role in plant defence, was also activated. The possible mechanisms by which Cu-induced growth delay and changes in enzymatic activities involved in plant defence processes are discussed. PMID- 14556384 TI - [Potential role of winter rape weeds in the extension of broomrape in Poitou Charentes]. AB - In the Poitou-Charentes district, among the 82 species of winter rape weeds identified, 22 displayed a strong affinity for this crop (Brassica napus L.). In fields, 50% of these weeds were parasitized by Orobanche ramosa, playing the role of host plants. Greenhouse co-cultures (weed/Orobanche ramosa) showed that weeds non-parasitized in fields could be attacked by broomrape, developing a more or less complete cycle. In vitro co-cultures (weed/Orobanche ramosa) revealed that root exudates of non-parasitized weeds, in fields or in greenhouse co-cultures, could induce Orobanche ramosa seed germination, but not attachment. These weeds could play the role of false hosts. PMID- 14556385 TI - [Multixenobiotic defense mechanism (MDMX) in bivalves]. AB - Multixenobiotic defence mechanism (MXDM) consists in a cellular system that functions as membrane extrusion pumps effluxing organic compounds out of the cells. In bivalves, it represents a primordial protection against toxic effects of organic xenobiotics in preventing their cellular accumulation. It has raised attention during the last decade for its potential to be used as a biomarker of pollution. This article reviews the fundamental knowledge on the MXDM system in bivalves and the methods proposed to assess its activity. Finally, it reviews the major results of laboratory and field studies that enabled to hypothesise that MXDM could be used as a biomarker of environmental stress rather than of pollutant exposure. PMID- 14556386 TI - Stem cells: ethics, legislation and regulation. PMID- 14556387 TI - Maternal food calling in domestic hens: influence of feeding context. AB - The aim of this study was to understand the relationship between production of food calls by maternal hens and food context. In a series of experiments with broody hens, we manipulated quality of items, quantity of food, food experience and dispersion of food items. We measured the frequency of food calling during standardized tests. Our results show that all the variables tested had significant effects on food calling. These results present some similarities and some discrepancies with previous reports on food calling by cockerels. PMID- 14556388 TI - Status of the so-called African pygmy elephant (Loxodonta pumilio (NOACK 1906)): phylogeny of cytochrome b and mitochondrial control region sequences. AB - Among the African elephants, it has been unanimously acknowledged that the forest elephants (cyclotis form) are peculiar, so that they have been elevated to the specific rank. The development of molecular analyses of extant Loxodonta has only focused on two forms yet: the savannah form (africana) and the forest form (cyclotis), disregarding the so-called pygmy elephants (pumilio or fransseni) the systematic status of which has been debated since their discovery. Therefore, we have sampled nine dwarfed-labelled specimens in collection and eight specimens of typical forest elephants that we compared to three savannah elephants and two Asian elephants. Because of the degraded nature of the nuclear DNA content in bone samples of old specimens, we assayed mitochondrial markers; 1961 bp of the mitochondrial genome were sequenced (over a continuous range spanning the cytochrome b gene, tRNA Thr, tRNA Pro, hypervariable region 1 and central conserved region of the control region). Pumilio and cyclotis are not sister taxa: the phylogenetic analyses rather account for the inclusion of the so-called pygmy elephants within a monophyletic group of forest elephants sensu lato. The internal structure of this clade reveals to depend on isolation and remoteness between populations, characteristics that may have been extensively influenced by climatic variations during the Quaternary period. We conclude that the specific taxon Loxodonta pumilio (or Loxodonta fransseni) should be abandoned. PMID- 14556389 TI - [Substitution therapy with buprenorphine to pregnant women with opioid dependency]. PMID- 14556390 TI - [Buprenorphine as opioid substitution to pregnant addicts]. PMID- 14556391 TI - [What factors are of significance in selection of hospital?]. PMID- 14556392 TI - [The use of the free choice of hospital rights in three different counties, 1991 1999]. PMID- 14556393 TI - [Evidence concerning umbilical cord hygiene in newborn infants]. PMID- 14556394 TI - [Substitutions therapy of opiate addiction in Denmark]. PMID- 14556395 TI - [Prehospital treatment of patients with opiate overdose in Copenhagen 1995-1998]. PMID- 14556396 TI - [Paternal inheritance of mitochondrial DNA]. PMID- 14556397 TI - [Tracheal rupture after orotracheal intubation]. PMID- 14556398 TI - [Hormone therapy and breast cancer]. PMID- 14556399 TI - [Pneumoconiosis--news about radiographic assessment]. PMID- 14556400 TI - [Street narcotics]. PMID- 14556401 TI - [Cochrane. The missing link?]. PMID- 14556402 TI - [A union basis of values stigmatize the mentally ill--there is a need for ethical analysis]. PMID- 14556403 TI - [Abbreviations and thrombosis prophylaxis one more time]. PMID- 14556404 TI - [The natural nutrition of human beings]. PMID- 14556405 TI - [Are sick people not allowed to go to work?]. PMID- 14556406 TI - Where quality is job one? PMID- 14556407 TI - Grandma, her shiny buick, and the marketplace. PMID- 14556408 TI - Brown Medical School: class of 2003. PMID- 14556409 TI - Is there a virtual medical school on the horizon? PMID- 14556410 TI - "It ain't easy being green," a case-based analysis of ethics and medical education on the wards. PMID- 14556411 TI - Protecting the public and assuring high practice and professional standards in the physician community: the Rhode Island Board of Medical Licensure and Discipline. PMID- 14556412 TI - Images in medicine. Dysphagia lusoria. PMID- 14556413 TI - Fickle syrinx. PMID- 14556414 TI - Free cancer screening for uninsured and underinsured women in Rhode Island. PMID- 14556415 TI - Utilization of hospital inpatient care in Rhode Island, 2000. PMID- 14556416 TI - Extracranial vertebral artery dissection. PMID- 14556417 TI - The myth of holism. PMID- 14556418 TI - Meaning and agency in discussing end-of-life care: a study of elderly veterans' values and interpretations. AB - The authors of this exploratory study used textual analysis of transcribed interviews to examine the mental constructs that individuals form around advance care terminology and to learn how elderly veterans conceptualize the language used in the Veterans Administration advance directive. They found that respondents often negotiated meaning by drawing on rigid schemas, specific mental constructs already in place: The Lord's Will, Machine Talk, Being a Burden, and Being Productive. The authors also examined the transcripts for agency. In addition to assigning external agency for end-of-life care decisions, respondents often expressed a complex interaction of "self" and "other" agency. These results challenge us to develop communication methods that allow patients to claim agency and participate fully in decisions regarding their health care, especially at the end of life. PMID- 14556419 TI - Identifying signals of suffering by linking verbal and facial cues. AB - Here, the authors describe microanalytically the two main behavioral states in suffering (enduring and emotional suffering) so that in subsequent research, appropriate comforting responses to ease and relieve suffering can be identified for each behavioral state. Their objectives were to describe the facial expressions of enduring and emotional suffering, and to link them with verbal narrative and thus develop a microanalytic description of each behavioral state. Using Ekman's modified EMFACS, they videotaped interviews with 19 participants and coded co-occurring verbal text and expressions. They also documented differences between each behavioral state and the transitions from enduring to emotional suffering. Enduring and emotional suffering are distinct and identifiable behaviors. These formerly implicit behavioral cues can be used in clinical assessment and research. PMID- 14556420 TI - Implicit exchanges in family caregiving for frail elders in Taiwan. AB - In this study, the authors use in-depth interviews with inductive analysis to develop a conceptual framework for exploring social exchanges and their implicit calculations for caregivers in Taiwan. They interviewed 12 caregivers, based on theory-based sampling and maximum variation. They found some components of implicit exchanges of the caregivers, and drew a framework to describe it. At the beginning of care, motivations were mostly from obligation accompanied by reciprocity or repaying motives. In the process of caregiving, some unique, implicit cultural implicit exchanges were found, such as karma, a demonstrative behavior to investment, equitable share of responsibility, and the pressure or rewards from public opinion. These implicit exchanges might be intermediary factors in helping caregivers cope with their burden or even in influencing their continuation of care. The findings are implicated to help family caregivers continue their care and not damage their quality of care. PMID- 14556422 TI - Understanding the barriers to cervical cancer screening among older women. AB - Despite the fact that 90% of cervical cancers can be prevented by regular Pap screening, many women are not screened regularly, particularly older women. To understand better the barriers to screening, the authors held five focus groups during April 2000 in Prince Edward Island (PEI), Canada. Of the 32% of women aged 45 to 70 who had not been screened over a 5-year period (1995-2000), 60 randomly selected women participated in this research. The authors analyzed discussions for themes using an inductive approach, with interpretation guided by the population health model. Women who have avoided a Pap test are falling between the cracks created by a complex interaction between personal experiences and the health system's approach to Pap screening. PMID- 14556421 TI - Participant experiences of Talking Circles on type 2 diabetes in two Northern Plains American Indian Tribes. AB - The Talking Circle, a culturally appropriate, 12-week educational intervention, was employed on two Northern Plains American Indian reservations to provide information on type 2 diabetes. In a phenomenological study, funded as a minority supplement to the Talking Circle intervention, the authors asked 8 American Indian participants of the Talking Circle to describe their experience of being an American Indian Talking Circle participant. Seven common themes describe the phenomenon of participating in a Talking Circle diabetic intervention. The Talking Circle technique was effective in providing information on type 2 diabetes through culturally appropriate community sharing. Type 2 diabetes is viewed by both outsiders and those involved as a chronic disease of the utmost concern in American Indian communities. PMID- 14556423 TI - Postpartum flatal and Fecal Incontinence Quality-of-Life Scale: a disease- and population-specific measure. AB - Using various recruiting methods, the authors identified 10 women who suffer from flatal and/or fecal incontinence subsequent to one or more previous vaginal deliveries. Each of these women participated in an individual in-depth 1-hour interview assessing symptom frequency, severity, and impact on quality of life. Participants also completed the Fecal Incontinence Quality-of-Life Scale and evaluated how well this scale captured their experiences. The authors used qualitative analyses to generate themes from the interviews and modified the existing scale, adding new items and themes to capture this population's particular symptom experience. This scale is being evaluated in the context of a surgical clinical trial comparing two techniques for repairing anal sphincter lacerations from delivery. PMID- 14556424 TI - "I wish he hadn't told me that": methodological and ethical issues in social trauma and conflict research. AB - Undertaking research on individuals who have experienced social traumas, such as being a victim or perpetrator of genocides and wars, presents difficult decisions for qualitative researchers. Deciding how to deal with these issues becomes more problematic when the researcher is a member of the society in conflict. To do this work, and to work collaboratively with researchers from the other side, sensitive ways to collect data have to be chosen. Interpretations of the materials can be no less difficult: Analyses often lead to information and understandings that may be difficult for the researcher to deal with from ethical, moral, and personal standpoints, especially when he or she is a member of the society and culture under study. In this keynote address, the author explores methodological and ethical issues connected to these topics. She brings examples from her work on the Palestinian-Israeli conflict and focuses on use of the life story methodology. PMID- 14556425 TI - Video-cued narrative reflection: a research approach for articulating tacit, relational, and embodied understandings. AB - The author's purpose in this article is to describe the effectiveness of video cued narrative reflection as a research approach for accessing relational, practice-based, and lived understandings. Video-cued narrative reflection provides moment-by-moment access to tacit experience. The immediate nature of the videotape captures emotional nuances, embodied perceptions, spatial influences, relational understandings, situational factors, and temporal manifestations. By watching videotaped interactions, participants are able to re-collect, re experience, and interpret their life world. Video-cued narrative reflection allows participants to be simultaneously engaged and reflective while describing significant understandings. By inserting audiotaped reflective commentary of participants into the original videotape transcript, contextual meanings can be located and articulated more easily. Although not appropriate for all types of research, this approach offers promise for certain studies. PMID- 14556426 TI - Validation of qualitative research in the "real world". AB - In this article, the author takes up the debate about the usefulness of the concept of validity in qualitative research and acknowledges the critical role of the researcher as an "instrument" in the research process. Qualitative research, and the process of analysis in particular, involves continuous reflexivity and self-scrutiny. Balancing the need for creativity and rigor, the qualitative researcher can experience uncertainty, particularly in relation to small numbers. The author describes steps that she and a colleague took to ensure the validity and accuracy of the findings in a qualitative study of female sex workers. She discusses specific challenges in relation to the validity of their interpretation and describes two unexpected and serendipitous validity checks that served as affirmation. PMID- 14556427 TI - Attaining the 3 x 5 goal: distributing antiretroviral therapy to 3 million by 2005. PMID- 14556428 TI - Pre-exposure chemoprophylaxis (PREP) as an HIV prevention strategy. AB - Chemoprophylaxis may be a prevention strategy for the sexual transmission of human immunodeficiency virus (HIV). Evidence suggests that condom use has waned with the availability of antiretroviral medication, at least in some resource rich settings. Barrier methods of HIV prevention have inherent problems, and the potential for failure. Microbicide research has focused primarily on male-to female transmission. Analogous to post-exposure prophylaxis, HIV prevention may be achieved by pre-exposure prophylaxis in some settings. Research in this potential strategy may be rewarding. PMID- 14556429 TI - Tuberculosis and HIV: a partnership against the most vulnerable. AB - Tuberculosis (TB) is a major cause of morbidity and mortality worldwide. Each year, there are eight million new Mycobacterium tuberculosis complex (MTB) infections and three million TB-related deaths. The catastrophic effects of TB are borne disproportionately among the most vulnerable. The HIV pandemic has further increased the burden so that the risk of TB reactivation from latency is 5 to 15 percent in HIV/TB coinfection. Tuberculosis reactivation fuels further primary infections, creating a vicious cycle of increasing infection, disease, and deaths. In addition, drug-resistant TB exacerbates this increasingly common problem. The clinical presentations of TB in relation to HIV and HIV-associated immune deficiency are discussed from the perspective of clinical diagnosis and treatment in patient care. Tuberculosis prophylaxis, concurrent drug treatment of TB and HIV, drug interactions, and overlapping toxicities are detailed for the practitioner. Immune reconstitution inflammatory reactions are now a common phenomenon in HIV treatment, where similar reactions have been less commonly described in TB treatment in the past. Global distributive injustices in wealth, the burden of disease, and the provision of healthcare are obvious in TB, and clearly show us that the needs of the most vulnerable populations must be met in order to address the problems. PMID- 14556430 TI - Pharmacological principles governing the use of proton pump inhibitors: tailoring therapy to improve GERD outcomes. Preface. PMID- 14556431 TI - Living with chronic heartburn: insights into its debilitating effects. AB - Heartburn is a common, often disabling condition. Twenty percent of adults exhibit symptoms at least once weekly. Few obtain complete satisfactory relief of their symptoms without the simultaneous implementation of significant lifestyle modifications and appropriate pharmacologic intervention. Poor sleep and chronic symptoms impair the QoL in 25% to 30% of such cases. Nocturnal symptoms are more troublesome, more difficult to treat, and are often manifested by extraesophageal symptoms. Several patients, however, unknown to their physicians continue to experience heartburn, despite lifestyle changes and taking prescription drugs. Adequate acid suppression is currently essential to effective management of the condition. PMID- 14556432 TI - The role of acid suppression in the management and prevention of gastrointestinal hemorrhage associated with gastroduodenal ulcers. AB - Peptic ulcer bleeding remains a substantial source of morbidity and mortality in the ICU setting. Endoscopic injection with adrenaline and thermocoagulation is the mainstay of treatment for peptic ulcer bleeds. To enhance healing and overcome limitations of endoscopic therapies, acid suppression therapy is recommended. Although results from a few studies do not support their use fully following an episode of acute UGI bleeding, PPIs have been used successfully to lower transfusion requirements and additional surgical procedures, reduce hospital stays, and lower medical costs. H2RAs and PPIs have a rapid onset of action when given intravenously; however, patients quickly become tolerant to the effects of H2RAs, generally requiring increased doses of medication after the first day of administration. PPIs provide persistent acid suppression, maintaining neutral gastric pH, especially during the critical first 72 hours following a bleed. Recent clinical studies further support their use in preventing bleeding in the clinical setting. Controversy exists over the utility of pharmacologically induced acid suppression in critically ill patients at risk for stress ulcers. Comparative pH studies, however, suggest that i.v. PPIs such as pantoprazole are more effective in raising intragastric pH than are H2RAs. Although the clinical benefits of PPIs for stress ulcer prophylaxis have not been established, there is a theoretical framework suggesting that they should be beneficial. Ongoing clinical studies should show whether the theoretical advantage translates into clinically meaningful benefits. PMID- 14556433 TI - Pharmacologic features of proton pump inhibitors and their potential relevance to clinical practice. AB - In conclusion, PPIs differ slightly in their PK and PD properties and in their propensity to interact with other medications. Inhibition of acid secretion is a function of proton pump inhibition, and PPIs work in a similar fashion to achieve this goal. PK/PD principles can be used to optimize PPI therapy when considering issues related to administration, such as timing of drug delivery relative to the number of active pumps and the drug's AUC. PMID- 14556434 TI - Managing gastroesophageal reflux disease: from pharmacology to the clinical arena. AB - GERD is a condition affecting patients throughout the 24-hour period, although the nighttime interval may require special consideration because of the pharmacologic profile of the agents used to treat GERD, and the normal physiologic processes rendering nighttime GERD particularly damaging. GERD patients should be managed with appropriate therapy proportional to the frequency and severity of their symptoms. PPIs are the most potent inhibitors of acid secretion, and with a thorough knowledge of their pharmacologic properties, clinicians can be helped in identifying strategies that can maximize the benefits of their potency (see Table 2). PPIs offer significant benefit to persons requiring longer-term therapy because they are potent agents and offer ease of dosing and favorable drug interaction and adverse effect profiles. However, it is necessary that clinicians understand the physiology and pharmacology of acid secretion to use them appropriately. Inevitably, proper therapeutic treatment demands that variables such as pharmacokinetics, ethnicity (metabolic profile), and the normal physiology of acid secretion be considered when choosing an appropriate PPI. PMID- 14556435 TI - Advances in the surgical treatment of colon and rectal cancer. AB - A family history of colorectal cancer is the basis for risk stratification and allows reliable screening recommendations for individual patients. It is also the starting point for diagnosing hereditary forms of colorectal cancer. The surgical implications of treating hereditary nonpolyposis colorectal cancer and familial adenomatous polyposis are briefly described. Laparoscopic colectomy and sentinel node biopsies are new techniques with potential application in the treatment of colon cancer. For rectal cancer, advances have resulted in better preoperative staging and improved oncologic and functional outcomes. Surgical intervention for recurrent colorectal cancer requires careful patient selection and a multidisciplinary approach. The evaluation and treatment of patients with different patterns of recurrent colorectal cancer is described. PMID- 14556437 TI - The forgotten one. PMID- 14556436 TI - Complications of cirrhosis. PMID- 14556438 TI - A cancer clip. PMID- 14556439 TI - Bigger is not always better. PMID- 14556440 TI - [Heterogeneity of Parkinson's disease]. AB - Parkinson's disease was, until recently, a unique disease. The discovery of several genetic factors has emphasized the heterogeneity of the disease. Analyse of structure and function of these gene products points to their critical role in dopaminergic neurons death and pathophysiology of parkinson's disease. In the mean time the clinical constellation of parkinsonian syndromes have been lumped into "synucleinopathies" and "taupathies" that both share common pathologic lesions. It is likely that clarification of the combined genetic and environmental factors undelying these various disorders will lead to novel diagnostic and therapeutic strategies. PMID- 14556441 TI - [Lewy bodies, a misleading marker for Parkinson's disease?]. AB - The Lewy body, an eosinophilic inclusion around 10 microns in diameter, is localised in the neuronal perikaryon. Its dense core is surrounded by a clear halo, which is lacking in the so-called "cortical Lewy bodies". Numerous proteins have been identified in Lewy bodies, among which the three neurofilament isoforms, ubiquitin and proteasome subunits. More recently, alpha-synuclein--a pre-synaptic protein--has been found to be the essential constituent of the Lewy body. Alpha-synuclein antibody has greatly increased the sensitivity of the neuropathological examination: it has emphasized the frequency of "Lewy neurites" (accumulation of alpha--synuclein in neuronal processes) and has shown the importance of extra-nigral pathology. Lewy bodies and neurites are indeed to be found in many areas of the central and peripheral nervous system: stellate ganglia, cardiac and enteric plexus, pigmented nuclei of the brainstem, basal nucleus of Meynert, amygdala, limbic nuclei of the thalamus, parahippocampal and cingulate gyri, insula and isocortex. Lewy body diseases include at least three clinical syndromes: 1) idiopathic Parkinson disease in which the brainstem bears the brunt of the pathology 2) Parkinson disease dementia in which Lewy lesions are found in the brainstem and are also abundant in the isocortex. A large number of senile plaques is frequently associated. 3) In dementia with Lewy bodies, the same lesions are observed but the cognitive deficit occurs first or shortly (less than one year) after the motor symptoms. PMID- 14556442 TI - [Brain imaging and mobility in Parkinson's disease]. AB - Numerous abnormalities of motor function have been identified in Parkinson's disease and functional imaging methods have given us a better understanding of underlying mechanisms. An underactivity of some frontal areas, in particular the supplementary motor area and the dorsolateral-prefrontal cortex, has been found in several studies. The activity of these areas increases when motor function improves following administration of dopaminergic drugs or functional neurosurgery. The dysfunction of the basal ganglia-thalamo-cortical loops in Parkinson's disease and in particular the excess of inhibition of the thalamo cortical pathway by the internal pallidum is likely to be involved in the abnormal activity of the cortical areas. An excess of activation has been demonstrated in other areas and is probably compensatory. PMID- 14556443 TI - [Therapeutic and physiopathological contribution of electric stimulation of deep brain structures in Parkinson's disease]. AB - Ablative functional neurosurgery constituted during the first half of the 20th century the main treatment of advanced forms of Parkinson's disease. The surgical procedure was spectacularly efficient on tremor, but sometimes complications were not totally regressive and even more severe when surgery was bilateral, this leading to neurocognitive deficits or to speech problems. This, associated to the efficiency of levodopa, was at the origin of the almost total disappearance of this type of surgery during the '60s until the eighties. The rebirth of functional neurosurgery, necessitated by the appearance of dyskinesias, was possible in most part because of the development of techniques with a lower morbidity, such as high frequency stimulation (HFS). This was initially applied to the thalamic ventral intermedius nucleus VIM, and the low morbidity of the method has been demonstrated by the possibility to operate a large number of patients bilaterally without complication. The demonstration that the pallidal target, which had been abandoned during the '50s because of its low efficiency on the triad of symptoms of Parkinson's disease, was selectively efficient on dyskinesias, led naturally to apply HFS to this target. In 1990, the demonstration in the field of fundamental research of the role of the subthalamic nucleus as a key element of regulation of movement, suggested to register this nucleus into the list of targets, despite the risk of hemiballism that this structure presents when it is lesioned by haemorrhage, because of the good tolerance of HFS as a surgical method. This target quickly showed its remarkable efficiency on all symptoms of Parkinson's disease. Because of the reduction of the doses of dopaminergic treatments which were allowed, this target had also the capacity to reduce, indirectly, the intensity of dyskinesias. If the efficiency of the method is nowadays demonstrated as well as the stability on the long term of its results, the mechanism is still mostly not understood. Moreover the physiopathogenic hypothesis suggests the possibility of a neuroprotective effect of the stimulation, which still needs to be clearly established at the experimental level as well as at the level of clinical applications. PMID- 14556444 TI - [Results of heart transplantation: experience of 233 grafts]. AB - BACKGROUND: Heart failure is one of the leading causes of hospitalization and death. The aim of this study was to evaluate long term outcomes after cardiac transplantation. METHODS: A retrospective review of 222 consecutive cardiac allograft recipients who underwent 233 transplantations between 1986 to 2000 was undertaken. Cardiomyopathy (123) and ischemic heart disease (87) were the most common indications. Mean age was 51 years +/- 11, and male gender was predominant (184). RESULTS: 33 patients (14.8%) died in the post operative period, mainly from graft failure (24 pts). During the follow-up period (total 1157 pt/yrs, mean 6.2 +/- 5 yrs, max 16 yrs), 60 late deaths occurred from cancer (21), graft failure (13), infection (13), and miscellaneous (13). Retransplantations were performed in 11 patients. The actuarial survival of the entire cohort was 75%, 66% and 50% at 1, 5, and 10 years respectively. CONCLUSIONS: Cardiac transplantation gives satisfactory long term results for patients with end-stage heart failure, providing good exercise tolerance and survival for 10 years or more in a large number of patients. Improvement in immunosuppression therapy is responsible for decrease in acute rejection rate. Reduction in HLA mismatch should allow for better immunomodulation and decreased incidence of cardiac allograft vasculopathy and malignancies. PMID- 14556446 TI - [Turner syndrome and mosaicism]. AB - Turner's syndrome is a gonadosomatic dysgenesis of female phenotype due to a more or less complete monosomy of one of the X chromosomes leading to a haploinsufficiency of the development genes situated at the level of the pseudoautosomal region of the gonosomes. Further experience of the karyotype showed a preponderance of mosaics and considerable variability of Turner's phenotype in proportion to the number of 45, X cells. The rare cases of monozygotism discordant with variable tissular distribution mosaics show that the phenotypic expression is a genic dosage effect. In patients with TS it would thus be of interest to study a second tissue such skin fibroblasts when a discordance is observed between the phenotype and the karyotype. PMID- 14556445 TI - [Repair of the mammalian central nervous system: the "spinal cord" model]. AB - The central nervous system of adult mammals has been classically considered as structurally rigid, tightly wired, and unable to be repaired. We have shown that there exists a rather considerable degree of intrinsic plasticity due to the neurons themselves, but merely to glial cells and to multipotent stem cells. The spinal cord constitutes a good model on which we could demonstrate, with vascular and traumatic animal paradigms, that an early pharmacologic intervention could reduce significantly the extent of lesions and the subsequent functional deficit. Moreover, we showed that regeneration of severed central axons could occur, provided that the astrocytes' component of the glial scar was modified. Finally, transplants of embryonic neurons were shown to repair the axonal circuitry below a sectioned cord, and to restore reflex functions. All these data point to unprecedented perspectives of efficient therapies in acute and chronic neurological diseases. PMID- 14556447 TI - [Breast reconstruction using a rectus abdominis myocutaneous flap: analysis of 234 cases]. AB - The pedicled rectus abdominis myocutaneous flap is a very sophisticated form of breast reconstruction. Provided the indications are carefully set out and a perfect mastery of the technique, the rate of complications is consequently very low (9 reinterventions for 234 cases: 3.8%). The authors stress one particular point which they deem important: the preliminary vascular preparation to improve arterial flow and venous drainage of the internal mammary pedicled. The high rate of good results and of patient satisfaction--84% in the reported series--makes this form of reconstruction the prime method to choose in the case of patients with different types of carcinoma and more particularly in the early stages of breast cancer or when there is a need for prophylactic mastectomy. PMID- 14556448 TI - [Importance of the nerve biopsy for the diagnosis of atypical forms of chronic inflammatory demyelinating polyradiculoneuritis: 8 cases]. AB - The objective of the study was to define how could be helpful a nerve biopsy for identification of atypical cases of chronic inflammatory demyelinating polyneuropathy (CIDP). An ad hoc committee in 1991 defined the clinical, electrophysiological and pathological criteria for diagnosis of CIDP. In common with other authors, we regard the rather specific electrophysiological criteria as being too restrictive, and we think that a significant number of patients may therefore not benefit from effective treatment or be excluded from therapeutic trials. The Inflammatory Neuropathy Cause and Treatment (INCAT) group (2001) has proposed new electrophysiological criteria of CIDP, which are more sensitive and do not loose any specificity. Over a period of three years (January 1999 to December 2001), we classified 44 patients into two categories: those presenting the strict criteria of the ad hoc committee and those who we regarded as cases of CIDP who did not meet these strict criteria. All these patients benefited from one or more clinical and electrophysiological examinations; extensive biological workup and genetic study when appropriate excluded other causes of neuropathy. Nerve biopsies were taken from all patients and samples were included in paraffin and epon for systematic light and electron microscopic examination. Out of 44 patients, 24 fulfilled the INCAT electrophysiological criteria with only 12 of these cases fulfilling the criteria of the ad hoc committee. Eight patients did not fulfill any of the widely accepted electrophysiological criteria of CIDP. However, study of nerve biopsies of these eight patients revealed histological features characteristic of CIDP according to histological criteria (AAN-1991). Among these patients, six have been treated and five responded favorably to conventional treatments for CIDP. Without information from the nerve biopsy, these patients would not have been treated effectively because their electrophysiological profile was indicative of axonal impairment interpreted erroneously as primary. PMID- 14556449 TI - [Intensive treatment of multiple myeloma]. AB - High dose therapy with autologous stem cell transplantation (ASCT) has been extensively used in the past 15 years in multiple myeloma. The IFM 90 trial has shown that autologous bone marrow transplantation (BMT) is superior to conventional chemotherapy in terms of response rate, event free survival, overall survival. Several other randomized studies confirm that ASCT yields superior complete remission and event free survival rates. However, the benefit for overall survival is not always significant because some patients may receive high dose therapy at the time of relapse. While ASCT appears to be the treatment of choice for younger patients, a number of questions have been addressed in the past few years (optimal conditioning regimen, best source of stem cells, impact of tandem autotransplants, role of maintenance therapy, results of transplantation in patients over 65 years of age or with renal failure). These issues are addressed in this review. Analysis of large cohorts of patients indicate that a low beta 2 microglobulin level and the absence of chromosome 13 abnormalities are associated with a better outcome. However in patients with a high beta 2 microglobulin level and chromosome 13 abnormalities, the prognosis is poor even after tandem transplantations. Allogeneic BMT is offered to a minority of younger patients with an HLA identical sibling. Initial series have shown a high-toxic death rate and no survival advantage compared to ASCT. Yet, allogeneic BMT is possibly the only curative therapy. Reports of CR achieved after infusion of donor lymphoid cells in patients relapsing after allogeneic BMT support the concept of a graft versus myeloma effect. Therefore, the objectives of current studies are to reduce transplant related mortality by using earlier BMT, better selection of patients, better graft-versus host prophylaxis or non myeloablative conditioning regimens. PMID- 14556450 TI - [Central nervous control of erection]. AB - Penile erection is caused by a change of the activity of efferent autonomic pathways to the erectile tissues and of somatic pathways to the perineal striated muscles. The sympathic outflow is mainly antierectile, the sacral parasympathic outflow is proerectile and the pudendal outflow, through contraction of the perineal striated muscles, enhances an erection already present. Spinal neurones controlling erection are activated by afferents from the genitals. It is likely that these primary afferents do not directly stimulate the spinal sympathetic, parasympathetic and somatic nuclei, but do through spinal interneurones. This spinal network is able of integrating information from periphery to elicit reflexive erections. It also receives supraspinal descending pathways from pons and hypothalamic nuclei, among the latter, the paraventricular nucleus and the medial preoptic area. These structures are likely involved to regulate penile erection in more integrated and coordinated responses occurring during sexual behavior. By receiving ascending projections from the spinal level which convoy informations from genitals, they could reinforce penile erection. Role of putative neuromediators or regulatory peptides is evoked. PMID- 14556451 TI - [Epidemiology of venous thrombosis]. AB - Venous thrombo-embolic disease (VTE) is a frequent pathology, responsible for around 10,000 deaths each year in France. Post-thrombotic sequellae is also a main problem related to the disease. The epidemiology of acquired and genetic risk factors of VTE is better understood and has allowed to show that in many cases it is a multifactorial disease. The association of several environmental or genetic and their analysis in the physiopathology of the disease and in the risk of recurrency is important in order to allow a better prevention of the disease and of its recurrencies. PMID- 14556452 TI - [Platelets and arterial thrombosis]. AB - The pathological mechanisms involved in arterial thrombus formation are similar to the mechanisms involved in physiological hemostasis. Arterial thrombosis is initiated following lesion of the vessel wall, either through rupture of an atherosclerotic plaque containing lipids, adhesive proteins and tissue factor or after angioplasty exposing the thrombogenic subendothelial matrix. Platelets play a major role in arterial thrombus formation through ADP secretion and thrombin generation on their activated surface. Arterial thrombosis is a frequent complication of atherosclerotic lesions and leads to acute ischemic events. These events are therapeutic emergencies which require administration of antithrombotic drugs inhibiting platelet functions and thrombin. PMID- 14556453 TI - [From heparin to synthetic antithrombotic oligosaccharides]. AB - In the early eighties, following breakthroughs in oligosaccharide chemistry, we achieved the total chemical synthesis of pentasaccharides related to the antithrombin-binding domain in heparin. Selective inhibitors of coagulation factor Xa, thus obtained, represent a new class of efficient antithrombotic drugs. In a further step, we have designed and synthesised oligosaccharides (pentadeca--to eicosasaccharides), comprising an antithrombin binding domain prolonged by a thrombin binding domain. These compounds inhibit both factor Xa and thrombin, in the presence of antithrombin, while they are devoid of undesirable non-specific interactions, particularly with platelet factor 4 (PF4). The efficacy of these new synthetic antithrombotics, with a unique biological profile, will be evaluated in clinical trials. PMID- 14556454 TI - [The Choay domain -- the structure responsible for the anticoagulant action of heparins]. AB - We describe the common structural basis for the anticoagulant action of the many different heparins available to the clinician. From different types of heparin we prepared fractions of virtually single molecular weight. We determined the molar concentration of material (HAM) containing the antithrombin (AT) binding pentasaccharide (A-domain), the specific catalytic activity in thrombin- and factor Xa inactivation and the capacity to inhibit thrombin generation (TG). We also calculated the molar concentration of A-domain with 12 sugar units at its non-reducing end, i.e. the structure that carries anti-thrombin activity (Choay- or C-domain). The anti-thrombin activity and the effects on TG are determined by the concentration of C-domain and independent of the source material or Mr. High Mr fractions (> 15,000) are less active, probably through interaction with non-AT plasma proteins. Anti-factor Xa activity is not indicative of anticoagulant potency but is a sensitive indicator of Mr and therefore predicts favourable pharmacokinetic properties: long half-life in the circulation and high bioavailability. PMID- 14556455 TI - [From fibrinogen to fibrin and its dissolution]. AB - Fibrinogen is a protein synthesised by the liver. It is converted by thrombin to an insoluble fibrin network to induce, together with platelet aggregates, haemostasis in response to rupture of endothelium. This change includes several steps and implied factor XIII. Molecular properties of fibrin are responsible for its important role in hemostasis which goes beyond the one of a simple final inert product of coagulation. In fact, fibrin regulates thrombin and factor XIII activities and its own destruction also called fibrinolysis. The multiple domains of fibrinogen and fibrin confer a role not only in haemostasis but also in wound healing, cellular migration and proliferation, due to interactions with endothelial cells, leukocytes and components of the extracellular matrix. Fibrin must be removed once its haemostatic role has been reached. The fibrinolytic process takes place in the vessel lumen. It is strongly regulated by the plasma concentration of an inhibitor called plasminogen activator inhibitor-1 (PAI-1) which synthesis strongly increases in obese insulin resistant and diabetic patients. Data from animal models show that increased PAI-1 production represents a prothrombotic state. Fibrinolysis plays also a role in tissue remodeling (vascular wall, placenta, adipose tissue....) by degrading the extracellular matrix, by activating growth factors or modifying cellular adhesion and migration properties. It has been proposed that PAI-1 in excess could be directly responsible for the development of atherothrombosis in insulin resistant subjects. Moreover recent results from transgenic mice indicate that PAI-1 in excess interferes also with weight gain. These data point out the importance of the haemostatic system in the extra vascular phenomenon of tissue remodeling. PMID- 14556456 TI - [Treatment and prevention of venous thromboembolic events: present and future antithrombotic agents]. AB - Two new classes of anticoagulants are actually developed which would change in the near future our strategies for the prevention and the treatment of venous thromboembolic events. These two classes are the anti-factor Xa and anti-factor IIa (direct antithrombin) agents. Among the anti factor Xa, the pentasaccharides are initiating their clinical use. Fondaparinux is a synthetic form of the natural pentasaccharide, its pharmacokinetics allows one s.c. administration/24 hours. It is active in prevention and treatment of venous thromboembolic and coronary thrombotic events. A modified form (idraparinux) whose pharmacokinetics allows one administration only once a week should have the same type of efficacy. Among direct antithrombin agents, hirudin and derivatives have been developed in the past decade with a limited use due to several drawback. More recently synthetic direct antithrombins modified to allow oral route have been developed, the most advanced in development, melagatran, is active in the prevention and treatment of venous thromboembolic and coronary thrombotic events. It could allow (if confirmed by clinical trials) a complete oral treatment of deep vein thrombosis without any biological monitoring. Melagatran is also active in the prevention of arterial thromboembolic events on atrial fibrillation. But other molecular forms of synthetic orally active direct antithrombin are also in development. Besides these important changes in our therapeutics which should appear in a near future, molecules aimed at other target are also tested: the most advanced are those antagonizing the initial phase of tissue factor activation of factor VII but other strategies are being tested such as stimulation of fibrinolysis. These new drugs at our disposal to treat venous thromboembolism should modify completely our handling of the patients. But additionally the numerous clinical trials necessary to prove the efficacy of the drugs, modify our understanding in the implication of the coagulation and in the physiopathogeny of thrombotic events. PMID- 14556457 TI - [Familial adenomatous polyposis: long term follow-up of patients after surgical treatment. Prognostic factors]. AB - The purposes of this study are: 1) to evaluate if recent progresses (knowledge of natural history, genetic diagnosis and surgical treatment) have an impact upon the long term follow up of familial adenomatous polyposis (FAP); 2) to assess the prognosis factors that are relative to recent progresses in diagnosis and treatment procedures. A retrospective study of 51 cases was carried out in July 2002 to analyse the following characteristics: phenotype, treatment, operative mortality and morbidity, late complications, especially rectal stump cancer after ileo-rectal anastomosis, duodenal adenomatosis and desmoid tumors. Twenty seven men and 24 women underwent surgery: 11 colo-rectal cancers were present at first step. Initial surgical procedures included 39 total colectomies with ileo-rectal anastomosis (IRA), 6 coloproctectomies with ileo-anal anastomosis (IAA) and 6 coloproctectomies with permanent ileostomy. Operative mortality was nil. Operative morbidity affected 11 patients. The rectum had to be secondary removed in 11 patients with convert in IAA. Duodenal adenomatosis required surgery in 10 patients: 5 surgical local excisions, 4 duodenopancreatectomies and 1 palliative by-pass. Six desmoid tumors were noted during the follow-up. On the whole 3 patients were lost of sight. Nine patients died (19.1%), 4 deaths were in relation with the disease: 1 rectal cancer, 2 duodenal cancers, 1 desmoid tumor necrosis. At the end of the follow up (mean duration: 17 years) 26 IRA and 17 IAA are present with good functional results. This study, according to already published data, suggests that today the risk of death related to colorectal cancer is becoming lower than the risk of death from duodenal cancer and desmoid tumor evolution, particularly since the introduction of the restorative proctocolectomy. The genetic diagnosis is useful in order to determine the choice of surgical procedures. PMID- 14556458 TI - [Serotonin and the cardiovascular system: role of the serotoninergic 5-HT 2B receptor]. AB - Identification of factors controlling myocardial differentiation and proliferation is of great importance for understanding the pathogenesis of congenital heart diseases. The molecular mechanism by which serotonin (5 hydroxytryptamine, 5-HT) regulates embryonic development of heart and cardiovascular functions remained unknown until recently. Inactivation of the 5 HT2B receptor (5-HT2BR) gene leads to embryonic and neonatal death due to the following defects in the heart: (i) 5-HT2BR mutant embryos exhibit a lack of trabeculae in the heart and a reduction in the expression levels of a tyrosine kinase receptor, ErbB-2, leading to mid-gestation lethality. These in vivo data suggest that the 5-HT2BR coupled to the heterotrimeric G protein Gq uses the signaling pathway of ErbB-2 for cardiac differentiation. (ii) Newborn 5-HT2BR-ko mutant mice exhibit cardiac dilation resulting from contractility and structural deficits at the intercellular junctions between cardiomyocytes. (iii) In adult 5 HT2BR mutant mice, echocardiography and electrocardiography confirm the presence of left ventricular dilation and decreased systolic function. These results constitute the first genetic evidence that 5-HT, via the 5-HT2BR, regulates differentiation and proliferation during development as well as cardiac structure and function in adults. PMID- 14556459 TI - [To make good use of medical error]. AB - Error, even if not made through negligence, always affects the physician. That probably explains why the literature on medical error is so sparse. Thanks to large surveys carried out mainly in American hospitals, awareness of the major role of this risk factor begins to appear. These studies demonstrate: three important facts: 1/ The frequency of errors with serious consequences is nearly 3% of all hospitalizations. 2/ Because of the growing complexity of modern health care system, medical errors have new characteristics. Even if they apparently originate from individual behaviour, they are in fact frequently rooted in the health care organisation and so are in fact systemic errors. 3/ A lot of these systemic errors could be prevented or corrected by measures at the level of the organisation. The study of mistakes is vital in implementing quality management of health care. Consequently it is the responsibility of the organisation to record and analyse adverse events. Each error can be an opportunity for safety improvement, provided it is acknowledged. PMID- 14556460 TI - [Degenerative neurological diseases of the central nervous system related to genetic neurolipidoses]. AB - Genetic neurometabolic diseases in childhood are multisystemic. Surprisingly, these genetic diseases can manifest for the first time during adolescence and adulthood. In this case, the clinical presentation and evolutivity are very different. In childhood, many neurological systems are touched and their evolution is rapidly lethal. In the adult, their presentation may be that of a degenerative disease of the central nervous system and, according to the disease, the syndrome is very particular and very systematized. From our clinical and biological experience, we would like to suggest a decision tree. PMID- 14556461 TI - [Spiral CT: is it time for lung cancer screening?]. AB - Recent studies have benefited from the progress of the scanner in order to relaunch the debate on the opportunity of screening lung cancer. Even if these initial studies have succeeded in showing that the practise of helical scan allows the screening of a considerable proportion of cases at an early stage which may have 5-year survivals of up to 70%, there is no specific indication of the diminution of lung cancer-specific mortality. Studies of cohorts of several dozen millions subjects during a period of 8 to 10 years are to be realised and an international pool will be charged of the harmonization of the protocols in order to realize the analyse of the collected information. An experimental study of 1000 subjects financed by a grant have started recently in France. PMID- 14556462 TI - [The law of medical responsibility. New stakes]. AB - The law of March 4, 2002, concerning the rights of patients and the quality of cares, establishes an extra jurisdictional way to solve conflicts of medical responsibility. After reminding the rules of common law which apply to this type of litigation, the text explains the outline of law goals, means used and established procedures. A medico-juridical critical study highlights confusions, imperfections and unvoiced comments of the text. Corrections might be necessary. The gravity of the stakes for medicine is underlined. PMID- 14556463 TI - [Body-contouring surgery]. AB - Concepts of beauty have been continuously evolving throughout the history of mankind. The voluptuous figures that were idealized by artists in the past have been substituted by slimmer forms. Medical advances in this century have permitted safe and efficient surgical correction of contour deformities. Until recently, these alterations were mostly hidden under heavy clothing or were reluctantly accepted. Current fashion trends generally promote body-revealing attire. The media frequently encourages the importance of fitness and good health linking these qualities with youthfulness and beauty. The subliminal as well as overt message is that these are necessary and desirable requirements for social acceptance and professional success. On the other hand, current sedentary lifestyle and dietary excesses, associated with factors such as genetic determination, pregnancy and the aging process, contribute to alterations of body contour that result in the loss of the individual's body image. This creates a strong psychological motivation for surgical correction. Localized fat deposits and skin flaccidity are sometimes resistant to the most sincere efforts in weight loss and sport activities. This ever-increasing request for contour surgery has been favorably met by safe and effective anesthesiology as well as efficient surgical techniques, resulting in a high degree of patient satisfaction. It is essential that today's aesthetic surgeon understand the motivations of patients who present with body contour deformities. A request for surgical treatment should be seen as a legitimate desire to achieve a physical form that approximates the individual with his or her ideal self-image. Additionally, the surgeon must always consider the possible benefit of including the participation of a multidisciplinary team approach. Depending on each case, this team should include consultants in endocrinology, dermatology, oculoplastics, pediatrics and other appropriate specialties. PMID- 14556464 TI - [New directions in the surgical treatment of primary hyperparathyroidism]. AB - For a long time, bilateral cervicotomy with a scrupulous exploration of the sites of the parathyroid glands was the technique of choice in the treatment of primary hyperparathyroidism. Recently new less invasive surgical techniques appeared: unilateral approach under local anaesthesia (UA LA) and video endoscopic parathyroidectomy (VEP), which could be used in 50% of patients after elimination of contraindications. Three factors support these new techniques: HPT I is in 85% of the patients related to a solitary adenoma. Ultrasonography or scintigraphy isolated or associated can detect the adenoma. Peroperative monitoring of the PTH can control the success of surgery. In our experience, 95 to 98 per cent of patients are cured of their HPTI.... In our opinion, UALA, technically simpler, with long term good results, is the technique of choice. PMID- 14556465 TI - [Treatment of prostate cancer using real-time brachytherapy]. PMID- 14556466 TI - [Critical analysis of the laparoscopic approach in colorectal surgery based on the personal experience of 613 interventions]. AB - Initial experience of colo-rectal laparoscopic surgery, after a period of enthusiasm tempered by anticipated technical difficulties, was held back due to reservations made in the case of operations for cancer, following the publication warning surgeons concerning the danger of the development of port-site metastasis. Nevertheless, this type of surgery continued to be developed for benign diseases of the colon, enabling acquiring of the necessary surgical skill. This skill could then applied by experienced surgical teams for the treatment of cancer, that do not seem to show negative aspects, in connection with local recurrences or 5 years survival rates. These reports need to be confirmed by ongoing multi-center trials currently being undertaken in several countries. The advantages of laparoscopic surgery in terms of post-operative comfort having been widely demonstrated, it seems that as applied strictly to colo-rectal pathologies, morbidity and mortality of this surgical procedure are at least equal to those of conventional "open" surgery. These good results require, however, a relatively long period of training. Based on the experience of 613 patients operated by laparoscopic surgery, the authors highlight the special characteristics of this surgical technique whose future expansion can be expected and is justified. PMID- 14556467 TI - [Spinal instrumentation, source of progress, but also revealing pitfalls]. AB - The second half of the XXo century and especially the last 30 years have been the source of a great improvement for surgical treatment of spinal pathology essentially in 3 directions:--First, for the patient himself and his comfort by suppression for most of the cases of any post operative external support thanks to the rigidity, security and strength of segmental fixation given by the hooks, screws and rods systems, as well for posterior as anterior instrumentation. In addition, these new techniques allow the patient to return quickly to standing and walking activity and subsequently the surgery for adult people increased dramatically especially for all kind of degenerative diseases and more and more extended spinal deformities.--The second major improvement came from the real and new understanding of the 3 dimensions for all the physiology and pathology of the spine leading to practical applications for the design and surgical strategies for correction. The exploding expansion of the era of computer technology brought a lot of help in such understanding as well as for the development of spinal instrumentation.--Finally the impressive development of medical imaging with CT scan and less and less invasive techniques like MRI allow a much better vision of spinal cord and roots (a major concern for the spinal surgeon). All this occurs also because simultaneous revolution occurred in the field of anesthesia and intensive care especially post operatively, but also because the big progress for monitoring of vital function as well as neurological monitoring during surgery. The consequence of that was an improvement for the results concerning the patient for functional quality of life as well as for cosmesis. The subsequent failures resulting of these improvements came from various fields even if we exclude infection or neurological complications more and more controlled now.--At the level of the indications because of these lack of post operative external immobilization, indication for cosmesis extend widely including adults. But also because these adult peoples are asking more and more for an active way of life including often sports. The indications for function also extend requiring more and more pre operative checking to limit the increase of the risk.--At the level of the pre operative strategy, most surgeons were establishing the strategy looking only on more or less close similar cases already done. In fact no personalized strategy done on biomechanical static and dynamic data of the patient as well as its connective and bone tissue proper qualities were really performed. It is why it is still now difficult to know pre operatively the behavior of the not fused area above and mainly below the instrumented area. Because the power of correction was very much improved with the new instruments, we can get some 3D imbalance immediately after or secondary acquired after such surgery as well from anterior or posterior devices. It is why we need pre operative simulation of surgery thanks to the computer.--At the level of surgery itself: it is not entered in the practical field any per operative 3D reliable measurement to know exactly the surgeon is doing because the maximum of reduction is not the optimum for many cases and this has to be quantified. The last thing is of course the difficulty to translate the lying position of the patient during surgery to the standing functional one. Some answers or perspectives for the future may come from:--The real pre operative simulation of the surgery (including levels of instrumentation amount of correction in 3D) according to the personalized values, stiffness, quality of soft tissues, weight, size, of the patient, localization and type of the deformity, etc.... Computer software already exists or are on the way to be expanded.--Per operative 3D control of what is doing the surgeon are also emerging with real time information in order to adjust the correction according to the pre op. simulation. In addition the noticed improvements on the biological field for fusion with bone activators like BMP, Hydroxyapatite, or bone substitutes like bio-active ceramics will probably help for fusion and decrease necessity of bone grafting. It is the same about disc regeneration which is on the way. Finally it is evident that the first steps already done for spinal surgery avoiding fusion will extend.--For children and growing spine, the challenge is major, but with memory metal instruments, laser precise destruction of abnormal growing structures as well as posterior flexible instrumentation avoiding stripping of the periosteum and leaving integrity of the disc and facet joints function, improvements are also on the way.--For adult and degenerative spinal deformities and pain, the development of spinal arthroplasty already done for the disc replacement will improve as well as for the posterior joints units where artificial ligaments experience will be replaced by real artificial joints still on experiment. In conclusion, some general biological medical questions are still waiting for answers:--Neurology and erect posture- Growth and degeneration--Malignancy (comprehension and control)--Pain and suffering. And of course what is the fact of the genetics for all of these problems: plenty of work for the future. PMID- 14556468 TI - [Evolution of the concept of mitochondrial disease]. AB - The concept of mitochondrial disease originated in 1962 when Luft and co-workers described a patient with non thyroidal hypermetabolism related to loose coupling of oxidation-phosphorylation in muscle mitochondria. Over the following quarter of century, with the routine use of the Engel-Gomory staining on muscle biopsy revealing ragged-red fibres as a convenient markers for mitochondrial pathology, numerous papers described clinical, biochemical and morphological aspects of mitochondrial encephalomyopathies. With the discovery in 1988 of mutations in mitochondrial DNA, the concept of mitochondrial disease with maternal transmission led to an explosive expansion of research in the field. Throughout the 1990's the rapid identification of multiple mitochondrial gene defects associated with clinically diverse disorders has left practitioners puzzled about diagnosing such heterogeneous and complexes syndromes. The great complexity of the system and the ubiquitous repartition of mitochondria explain the wide variety of clinical phenotypes associated with primary mitochondrial diseases due to mutations in the mitochondrial genome, in the nuclear genome or in the cross talk between the two genomes and regulations. In the past few years, the pivotal role of mitochondria in drug sensitivity, their key role in aging, apoptosis or neurodegeneration lead to a mitochondial medicine. Here the term of mitochondrial disease is limited to genetic defect in the respiratory chain where advance were recently especially significant for the evolution of the concept and updated classification. PMID- 14556469 TI - [Ethics and animal experimentation. The ethics committees in France and foreign countries]. AB - Functions, main features, organization and operating principles of ethical committees for animal experiments in France and foreign countries are presented from a general viewpoint. Four majors concepts are underlined: awareness of respect of animal, competence, open door policy, communication. PMID- 14556470 TI - [The state and health insurance]. AB - The relationship between the State and the health insurance passes through an institutional and financial crisis, leading the government to decide a new governance of the health care system and of the health insurance. The onset of the institutional crisis is the consequence of the confusion of the roles played by the State and the social partners. The social democracy installed by the French plan in 1945 and the autonomy of management of the health insurance established by the 1967 ordinances have failed. The administration parity (union and MEDEF) flew into pieces. The State had to step in by failing. The light is put on the financial crisis by the evolution of ONDAM (National Objective of the Health Insurance Expenses) which appears in the yearly law financing Social Security. The drift of the real expenses as compared to the passed ONDAM bill is constant and worsening. The question of reform includes the link between social democracy to be restored (social partners) and political democracy (Parliament and Government) to establish a contractual democracy. The Government made the announcement of an ONDAM sincere and medically oriented, based on tools agreed upon by all parties. The region could become a regulating step involving a regional health council. An accounting magistrate would be needed to consider not only the legal aspect but to include economic fallouts of health insurance. The role and the missions of the Social Security Accounting Committee should be reinforced. PMID- 14556471 TI - [The Law of 19 Ventose, year 21 (10 March 1803): fundamental text and provisional expedient]. AB - During the last years of the Ancien Regime, Fourcroy established the link between Vicq d'Azyr and Colombier who both wore themselves out in order to reform the medical profession, fallen into a state of turmoil. But both died, and henceforth everything was handled by Fourcroy. On 10 March 1803 (19 Ventose an XI) he arranged for the passage of a fundamental law--today is its bi-centennial- reorganising the profession. Henceforth no one could exercise that profession without having been received as a doctor, except in underdeveloped regions where "health officers" (less qualified practitioners) would be accepted. PMID- 14556472 TI - [Operability of intracranial meningiomas]. AB - Intracranial meningiomas are lesions which generally can be removed radically by surgery. However, because of their location, the extension of the dural attachment, the inclusion of the main arteries of the circle of Willis or of cranial nerves, total removal carries a high risk of morbidity. In a review of a personal series of 500 cases, the limits of surgery are discussed and different groups of tumors are defined corresponding to their surgical operability. Although surgery appears indisputable on the convexity of the brain, para sagittal and anterior basal meningiomas, total removal remains questionable for tumors of the inner part of the sphenoid ridge, of the petrous bone and of the falco-tentorial junction. In a few other cases, surgery is either unnecessary (asymptomatic meningioma) or inefficient in that there is no clear benefit for the patient (cavernous sinus meningioma). In the cases where surgery does not offer a complete cure, radiotherapy may be considered. PMID- 14556473 TI - [Surgical rehabilitation of the upper limbs after traumatic tetraplegia]. AB - Patients with a cervical spinal cord injury experience a complete paralysis of their lower limbs and a partial paralysis of their upper limbs associated with a dramatic decrease of upper limbs function. Twenty years ago, surgical rehabilitation of their upper limbs started to develop, based mainly on tendon transfer procedures. We report a consecutive series of 69 such patients operated in the past 12 years, amounting to 188 rehabilitation procedures on their upper limbs. Surgery was beneficial in all cases, there was no case of functional loss, and the patients were mostly satisfied with the procedures. PMID- 14556475 TI - [Recent data on the physiopathology of hyperthyroidism]. AB - Cloning of the TSH receptor has led to marked progress in the understanding of the pathogenesis of hyperthyroidism. Genetic studies in familial cases of Graves disease have allowed to dismiss the TSH receptor as a candidate gene. Loci harbouring genes probably involved in the occurrence of Graves' disease have been described. Purification of native TSH receptor has allowed to study directly and to characterize the autoantibodies directed against this antigen. In cases of non immune hyperthyroidism the implication of activating mutations of the TSH receptor and of the a subunit of Gs has been described. PMID- 14556474 TI - [Adverse drug effects observed at French admissions departments and emergency services (Prospective study of the National Educational Association for Teaching Therapeutics and proposals for preventive measures]. AB - Various studies have shown that adverse drug events (ADE) are a substantial cause of hospital admissions. However, little is known about the incidence and severity of ADE resulting in hospital visits. To address this issue, we conducted a prospective survey in 10 primary care and emergency departments of French public hospitals. This study was performed over two periods of one week, one in June 1999 and one in December 1999, in primary care and emergency departments of five university hospitals and five general hospitals throughout France. Out of a total of 1,937 patients consulting, 1,562 were taking at least one drug during the previous week and were included for analysis according to the protocol. Altogether, 328 (21%; 95% confidence interval 19% to 23%) of these patients receiving at least one drug consulted because of an ADE. The sex ratio (M/F) was the same in both groups with or without ADE (1.04 vs 1.02, respectively, P = 0.83). Patients with ADE were older than those without (63.5 vs 54.8 years, P < 0.0001). Furthermore, ADR patients were more likely to have a higher severity score than no-ADE group (P = 0.019). The outcome seemed to be worse in patients with an ADE. The percentage of patients treated with 2 or more drugs and the number of drug exposures were significantly higher in patients with ADE than in those without (90.9% vs 75.0%, P < 0.0001, and 5.17 vs 3.82, P < 0.0001, respectively). The most frequent causes of visits for ADE-patients were digestive (n = 53, 16.2%), neurological (n = 52, 15.9%), cardiovascular (n = 49, 14.9%) and malaise (n = 49, 14.9%) events. In total, 410 drugs were incriminated in the occurrence of 328 ADE. The most frequently incriminated drug classes were (1) psychotropic agents, including anxiolytics and/or hypnotics, antidepressants and antipsychotics (n = 84, 20.5%), (2) diuretics (n = 48, 11.7%), (3) anticoagulants (n = 38, 9.3%), (4) other cardiovascular drugs (n = 63, 15.4%), and (5) analgesics, including non steroidal antiinflammatory agents (n = 57, 13.9%). The avoidability of ADE could be estimated by an external expert panel in 280 of the 328 cases. In 106 cases (37.9%), ADE was considered to be preventable because a contra-indication or a warning about drug use had not been respected. PMID- 14556476 TI - [Psychological and neuropsychological problems in multiple sclerosis]. AB - Neuropsychological investigations have demonstrated that cognitive disorders are common (about 60%) in patients with multiple sclerosis. 22 patients and 22 controls participated in the study with a review of literature. The cognitive dysfunction may be termed a subcortical white matter dementia. The hallmarks are: forgetfulness, reduced speed of information processing, impaired attention and slowness of thought processes, impaired ability to manipulate acquired knowledge. Psychiatric disturbance have also high prevalence: emotional or personality changes, depression. Pathological laughing and crying are classical but not well understood. This intellectual and emotional changes in multiple sclerosis are studied by adapted psychometric psychiatric examination. Correlation of magnetic resonance imaging with neuropsychological testing is now demonstrated. Total lesion score is the best predictor of cognitive deficits, cerebral atrophy and lesions of the corpus collosium also. Neuropsychological rehabilitation techniques and symptomatic treatments must be applied to patients with multiple sclerosis. PMID- 14556477 TI - [The remyelination phenomenon in multiple sclerosis]. AB - Multiple sclerosis is an inflammatory demyelinating disease of the central nervous system. Available treatments (immuno-modulators, immuno-suppressants) limit central nervous system inflammation and are only partially effective. Remyelination of naked axons becomes insufficient in most cases as the disease progresses. The reason for this repair deficit are many, including oligodendroglial, axonal and environmental factors. Understanding why remyelination fails is crucial for devising effective methods by which to enhance it. PMID- 14556478 TI - [Epidemiology of childhood cancer]. AB - The knowledge of the epidemiology of cancer in childhood is relevant of a strict methodology allowed by the activation of population-based specific registries, case-control and cohort studies. Descriptive epidemiology is a mean of survey and of public health, while analytic epidemiology contributes to define the role of genetic and environmental factors, along with their interaction. The collected data help indirectly to improve the care of the child with cancer and to a better understanding of the carcinogenesis. PMID- 14556479 TI - [Therapeutic progress in acute lymphoblastic leukemias and stage IV Burkitt-type lymphomas in children]. AB - Prognosis of stage IV Burkitt's lymphoma according to Murphy's classification and FAB L3 acute lymphoblastic leukemia was dramatically improved during the two last decades through a very intensive and short duration chemotherapy. At the end of the seventies the rate of cure was 30% in stage IV Burkitt's lymphoma with up to 25% blasts in the bone marrow, and there was no cure for cases with central nervous system involvement and FAB L3 ALL. The French Society of Paediatric Oncology (SFOP) was one of the international leading groups in treatment research in these diseases. Successive LMB protocols led to the international LMB96 protocol involving France, United Kingdom and United States of America. Currently, event free survival of all stage IV Burkitt's lymphoma and FAB L3 LAL is higher than 85%, and approaches 80% in the cases with central nervous system involvement and 90% in the cases without CNS involvement. Relapse cases, occurring always early, still have a very bad prognosis and remain a challenge for the future. PMID- 14556480 TI - [The Luys' body]. AB - In 1865, Jules Luys (1828-1897) described the 'accessory band of the superior olive' (red nucleus). In 1877, A. Forel completed the description and gave the name of Corpus Luysii (CL) to this grey sub-thalamic formation. In 1927, P. Martin's attribute the Hemiballismus to the destruction of the C.L. and specifies the function of the nucleus foreseen by Luys, which play a crucial role in the synthesis of automatic motor action. The new stimulation techniques of this nucleus in the treatment of Parkinson's disease would have opened a third life for the C.L. if its name hade not be changed into 'nucleus subthalamicus'. Jules Luys, a good anatomist, one of the pioneers in France of microscope and photography, ruined his reputation by his ramblings on hysteria and hypnosis which allowed the 'action of medications at distance', the 'storage of cerebral activities within magnetic crowns' and gave prominence to 'brain emanations'. PMID- 14556481 TI - Polymeric membrane electrodes for selective determination of methiaden. AB - Polyvinyl chloride matrix membrane ion-selective electrodes for the determination of methiaden hydrochloride based on the ion-pair of methiaden cation with tetraphenylborate anion were prepared using bis(2-ethylhexyl)sebacate, (electrode A), 2-nitrophenyl octyl ether (electrode B). 2-nitrophenyl dodecyl ether (electrode C) and 1-isopropyl-4-nitrobenzene (electrode D) as the membrane solvents. The electrodes exhibit near-Nernstian response in different concentration ranges, depending on the used membrane solvents. The basic analytical parameters of these electrodes were evaluated. The potentiometric titration method was used to determine methiaden hydrochloride with good precision and accuracy. PMID- 14556482 TI - Kinetics of hydrolysis of diltiazem hydrochloride in aqueous solutions. AB - The kinetics of hydrolysis of diltiazem hydrochloride in aqueous solutions at 313, 323, 333 and 353 K over the pH-range 0.4-9.7 has been investigated. The decomposition was followed by the HPLC method. The pH-rate profile was accounted for by the specific acid- and base-catalysed reactions and also by assuming spontaneous or water-catalysed decomposition of both dissociated and undissociated molecules of diltiazem. Various buffer substances were found to exhibit general acid and base catalysis of the degradation. Thermodynamic parameters of the reaction: energy and enthalpy of activation and the frequency factor for the specific rate constants were determined. PMID- 14556483 TI - Feasibility of the Ph.Eur. flow-through cell for dissolution testing of the compounded rectal suppositories containing indomethacin or sodium diclofenac. AB - Ph.Eur. and BP have introduced a dissolution apparatus for suppositories. Suitability of the apparatus for quality control of indomethacin or sodium diclofenac (100 mg) compounded suppositories was evaluated and the effect of the type of suppository base on dissolution profiles was studied. The fastest and most reproducible release profiles were observed for hydrophilic base (macrogols). More than 80% of the drug was released during 60 min, while after 350 min 18.5-50% of the total amount was released from lipophilic bases (Witepsol and Adeps solidus). The results demonstrate that slow and non-reproducible release occurs when the lipophilic suppository base does not melt. The feasibility of the test for the formulations, which do not melt during the procedure, is questionable. PMID- 14556484 TI - Antimicrobial studies of hydrazone complexes of Hg(II) and Fe(II) divalent metal ions. AB - The antibacterial, antifungal and antitubercular activities of Hg(II) and Fe(II) complexes of hydrazone were studied. All the complexes have been screened against Staphylococcus aureus, Salmonella typhi, Candida albicans, Aspergillus niger, Saccharomyces cerevisiae and Mycobacterium tuberculosis H37Rv and found to be more toxic than the parent ligand. The activity increased in the order [5-methyl 3-oximino-hexan-2-one-hydrazone]2 Fe(II) < [5-methyl-3-oximino-hexan-2-one hydrazone]2 Hg(II) < 3-oximino-hexan-2-one-phenylhydrazone]2 Fe(II) < [5-methyl-3 oximino-hexan-2-one-phenylhydrazone]2 Hg(II) for antibacterial and antifungal activity. PMID- 14556485 TI - Synthesis of new N-substituted cyclic imides with an expected anxiolytic activity. XXIV. Derivatives of N-hydroxy-1-methoxybicyclo[2.2.2]oct-5-ene-2,3 dicarboximide. AB - The preparation of a number of derivatives of N-hydroxy-1 methoxybicyclo[2.2.2]oct-5-ene-2,3-dicarboximide with an expected anxiolytic and antidepressive activity has been described. PMID- 14556486 TI - Synthesis of new N-substituted cyclic imides with potential anxiolytic activity. XXV. Derivatives of halogenodibenzo[e.h]bicyclo[2.2.2]octane-2.3-dicarboximide. AB - The preparation of a number of derivatives of dibenzo[e.h]bicyclo[2.2.2]octane 2.3-dicarboximide with an expected anxiolytic activity. PMID- 14556487 TI - Antioxidant properties of captopril in vitro. AB - This paper reports on the effect of captopril on the stability of unsaturated fatty acids in the evening primrose oil. The experiment was performed for captopril at concentrations of 0.05%, 0.1% and 0.2% in the samples of evening primrose oil at temperatures of 20 degrees, 40 degrees and 60 degrees C. The determination was performed on the 10th day of incubation. The results were compared to those obtained for analogous samples with octyl gallate, an antioxidant commonly used in oil systems, taken at the same concentrations and at the same temperatures. The changes in concentrations of the substances studied with time were monitored by gas chromatography. The results have shown captopril to be a stronger antioxidant than octyl gallate at all concentrations and temperatures used. PMID- 14556488 TI - Determination of immunological activity in vitro of some plant raw materials. AB - Aqueous extracts of five tannin plant raw materials have been studied for the survivals of thymocytes from the thymus of Balb-c mice. The immunostimulation activity was compared with a preparation from calf thymi TFX and phenolic acids: ellagic and gallic acids. It was found that only the Fructus Myrtilli extract has revealed a statistically significant influence on the survival of thymocytes. PMID- 14556489 TI - Influence of felbamate on the antinociceptive action of morphine, metamizol and indomethacin in mice. AB - The influence of felbamate (200 mg/kg or 50 mg/kg) on antinociceptive effect of morphine (10 mg/kg), metamizol (500 mg/kg) and indomethacin (10 mg/kg or 1.4 mg/kg) was investigated in a mouse model using the hot-plate test. All drugs were injected intraperitoneally. Felbamate was administered to mice 30 min before applying the analgesic drugs. Measurement of nociception was performed within 2 h after felbamate administration. The research studies were further conducted with a multiple (10 days) drug dosage. Felbamate in the dose of 200 mg/kg significantly increases the analgesic effect of morphine and weakens the effect of metamizol and indomethacin. Multiple administration of felbamate does not affect the activity of morphine and metamizol, but decreases analgesic effect of indomethacin. PMID- 14556490 TI - Studies on brain biogenic amines in methanolic extract of Cuscuta reflexa Roxb. and Corchorus olitorius Linn. seed treated mice. AB - The methanolic extract of both Cuscuta reflexa stem and Corchorus olitorius seed showed marked protection against convulsion induced by chemoconvulsive agents in mice. The catecholamines contained were significantly increased in the processed extract treated mice. The amount of GABA, which is most likely to be involved in seizure activity, was increased significantly in mice brain after a six week treatment. Results of the present study revealed that both the processed extracts showed a significant anticonvulsive property by altering the level of catecholamines and brain amino acids in mice. PMID- 14556491 TI - New possibilities of application of multifunctional polymers and polymer conjugates. AB - Present review provides examples of new applications of multifunctional polymers and polymer conjugates, i.e. polymer-active substance conjugates, polymer-protein conjugates, in pharmacy and medicine. PMID- 14556492 TI - Selenium as an anticancerogenic agent. AB - Selenium is an essential component of the intracellular antioxidant system, which defends the cell and the whole organism against oxidative damage and so from oxidative disease, especially from cancer. Selenium may be considered as an anticancerogenic agent, and a higher concentration of selenium in plasma may be a protective factor against human malignancies. It is also interesting to note that higher doses of selenium, applied to patients simultaneously with chemotherapy, may lower the toxicity of antineoplasm drugs without weakening the therapeutic effect. PMID- 14556493 TI - The synthesis and applications of 5-aminolevulinic acid (ALA) derivatives in photodynamic therapy and photodiagnosis. AB - A route has been developed to high-purity precursors, viz., ALA esters, to be used in photodiagnosis and photodynamic therapy. Hexyl, butyl and methyl 5 aminolevulinates are similar to the ALA acid in chemical stability and efficacy in producing the appropriate photosensitizer PpIX. Tests carried out on animal models showed the method based on the esters to be the more selective. PMID- 14556494 TI - Search for new lead structures in the isoxazole heterocyclic system. AB - Looking for active immunosuppressant, a series of substituted phenylamides of 5 aminomethinimino-3-methylisoxazole-4-carboxylic acid was obtained, which showed immunosuppressory activities in the in vitro and in vivo tests, comparable with that of cyclosporine A. Rentgenostructural studies of three most representative derivatives were performed and the molecular modelling of compounds, demonstrating most characteristic biological activities, was performed. In the next stage, quantum-chemical investigations were conducted in order to determine structure-activity relationships. PMID- 14556495 TI - The optimal way from laboratory to industrial facility. AB - The chance of success on the way "from laboratory to industrial facility" greatly increases with shortening of the time of the research and development (R&D) phase, upon the condition that the process is ready for implementation, i.e. elaborated completely in the form of a process design with positive conclusions regarding the implementation (1,2). The complexity of process designing and a typical course of realization of an investment in the chemical industry are described. Profitability, teamwork as well as creative and alternative way of solving problems have been suggested as the three rules to be followed when elaborating a chemical process (3). The phases and stages of sealing up are shown. It has been emphasized that the optimal timing of the research-design implementation course is to be reached by overlapping of following steps and minimizing of the number of mistakes owing to good organization of the teamwork of researchers, design engineers and manufactures--specialists in different fields. The use of miniplants and mathematical methods of designing of experiments as an effective solution for some technological problems e.g. recycling, hazards, differences in laboratory and technological solutions, validation, has been proposed. Examples of the long-term development of technology and conquering the market as well as a rapid financial success as a result of by-passing the patent rights have been shown. PMID- 14556496 TI - [West Nile fever]. PMID- 14556497 TI - [RNAi-mediated specific gene silencing and its application to medical treatments of viral infectious disease]. PMID- 14556498 TI - [Subacute sclerosing panencephalitis (SSPE): viral mutation and neuropathogenicity]. PMID- 14556499 TI - [Japanese encephalitis]. PMID- 14556500 TI - [Neuropathogenesis by cytomegalovirus]. PMID- 14556501 TI - [Anti-poliovirus response of neural cells]. PMID- 14556502 TI - [A personal view on 50 years of the Japanese Society for Virology]. PMID- 14556503 TI - [Hantavirus infection as a rodent borne zoonosis]. PMID- 14556504 TI - [For the prevention and control of pandemic influenza]. PMID- 14556505 TI - [Fight against AIDS--activities by WHO]. PMID- 14556506 TI - [Toward the control of herpes simplex virus infections]. PMID- 14556507 TI - [How can we improve the system to analyze HCV replication system?]. PMID- 14556508 TI - [Report of the 50th annual meeting of the Japanese Society for Virology]. PMID- 14556509 TI - [Pathology of human herpesvirus 8 (HHV-8)]. PMID- 14556511 TI - [7th international congress on AIDS in Asia and the Pacific in the era of massive infectious diseases]. PMID- 14556510 TI - [The mechanism of persistent infection and pathogenesis of Borna disease virus]. PMID- 14556512 TI - Effect of beta-endorphin and DAGO, a selective agonist of mu-opioid receptors, on the proliferative activity of lymphocytes. PMID- 14556513 TI - The heptapeptide SEMAX stimulates BDNF expression in different areas of the rat brain in vivo. PMID- 14556514 TI - Determination of myocardium viability on the basis of the spectra of laser induced fluorescence. PMID- 14556516 TI - Central mechanisms that control respiration in the pulmonate snail Lymnaea stagnalis: phasic coupling of lung ventilation to cyclic locomotion. PMID- 14556515 TI - Study of the mechanism of the cytostatic effect of picolinic acid on proliferating Tetrahymena pyriformis cells. PMID- 14556517 TI - Trekrezan as a modulator of hemato- and immunopoieses. PMID- 14556518 TI - Changes in systemic arterial pressure response to norepinephrine in rabbits during adaptation to cold. PMID- 14556520 TI - Chemosensory systems of fish as indicators of aqueous medium quality. PMID- 14556519 TI - M-cholinergic response of arterial pressure in rabbit small intestine blood vessels during cold adaptation. PMID- 14556521 TI - Coordinated expression of the genes Gus and Mup as a potential basis of the functional activity of the androgen-dependent pheromones of the house mouse (Mus musculus L.). PMID- 14556522 TI - Effect of the NO donor sodium nitroprusside and the NO-synthase inhibitor L-NOARG on the binding of [3H]quinuclidinyl benzylate by muscarinic cholinoreceptors of the rat cerebral cortex. PMID- 14556523 TI - The role of proteinases of the enteral microbiota in temperature adaptation of fish and helminths. PMID- 14556524 TI - Birds spread soil microarthropods to Arctic islands. PMID- 14556525 TI - Revision of phylogenetic relationships in the antilopinae subfamily on the basis of the mitochondrial rRNA and beta-spectrin nuclear gene sequences. PMID- 14556526 TI - Effect of brassinosteroids on the hormonal balance in wheat seedlings. PMID- 14556527 TI - Kinetin eliminates an enhanced photoperiodic sensitivity of PHYB-transgenic potato plants. PMID- 14556529 TI - Elementary behavioral acts of valve movements in mussels (Mytilus edulis L.). PMID- 14556528 TI - The role of salicylic acid in systemic resistance of tomato to nematodes. PMID- 14556530 TI - A new locality of early cretaceous mammals in western Siberia. PMID- 14556531 TI - N-benzyl and O-benzyl compounds: a new group of synthetic auxin biomimetics. PMID- 14556532 TI - Reproduction of polytocous mammals: the role of the mother-offspring integrative physiological system. PMID- 14556533 TI - Location of the Prox1 gene expression during newt lens and retina regeneration. PMID- 14556534 TI - Seasonal variation of the histochemical parameters of skin glands in the vole Microtus socialis paradoxus and their importance for the taxonomic status. PMID- 14556535 TI - Genetic regulation of inflorescence development in chrysanthemum. PMID- 14556536 TI - Association of Atlantic hydrothermal mytilids of the genus Bathymodiolus spp. (Mollusca: Mytilidae) with the polychaeta Branchipolynoe aff. seepensis (Polychaeta: Polynoidae): commensalism or parasitism? PMID- 14556537 TI - Changes in the extracellular matrix surface network during cyclic reproduction of proembryonic cell complexes in the Fagopyrum tataricum (L.) gaertn callus. PMID- 14556538 TI - Determination of the proteins and capsids of herpes simplex virus in human spermatozoa. PMID- 14556539 TI - [Antibodies to H+/K+-ATPase of gastric parietal cells in children with Helicobacter pylori associated chronic gastritis]. AB - 54 children at the age of 3-15 suffering from HP-associated CG were examined. Antibodies to H+/K(+)-ATPase of gastric parietal cells were determined with the help of the immune-enzyme analysis method. It was discovered that antibodies to H+/K(+)-ATPase are found in 27.7% of children. Concomitant autoimmune diseases, pangastritis, morphologic symptoms of the pre-atrophic stage of autoimmune gastritis as well as the drop of secretion at the fasting phase were discovered more often in antibody-positive patients. The successful eradication of HP in two children resulted in the disappearance of anti-parietal autoantibodies. PMID- 14556540 TI - [Status of antibiotic-resistant Helicobacter pylori in Russia]. PMID- 14556541 TI - [Current methods of ulcer therapy and their efficacy and cost]. AB - The purpose of this study is to assess the efficiency of the anti-ulcer therapy taking into account the economic aspects of treatment. We have examined 708 HP positive(+) patients with stomach ulcers (SU) and duodenal ulcers (DU). Various schemes of anti-helicobacter and anti-secretory therapy were administered to them. Positive dynamics of some clinical data (local palpatory tenderness and abdominal wall resistance) and gastritis (gastroduodenitis) activity was discovered in all groups of patients. The cost of the treatment was determined with the help of marketing studies by taking the lowest price of drugs. There were no statistically reliable differences found groups of traditional treatment regimens by both results of HP eradication and the cost of eradicative and anti secretory therapy. All drugs under study are highly effective for rapid relief of symptoms at an exacerbation of stomach ulcer associated with HP as well as the decrease of gastritis (gastroduodenitis) activity. It is possible to recommend them for both eradicative therapy and prolonged treatment. PMID- 14556542 TI - [Use of the immunomodulator hepon in the treatment of erosive peptic ulcer lesions]. PMID- 14556544 TI - [Problem of etiological multifactorial property of stomach cancer in current studies]. AB - The study of tumor growth morphogenesis is an urgent problem of modern oncology. The resolution of the stomach cancer problem is related to the study of those pathologic processes in both the organism as a whole and the mucous coat of the stomach in particular, which precede the malignant growth development. First of all, this is a question of pretumor diseases. PMID- 14556543 TI - [Use of omeprazole and esomeprazole in patients suffering from bronchial asthma with associated gastroesophageal reflux disease]. AB - The purpose of this study is a comparative assessment of the clinical efficiency of Omeprazole and Esomeprazole in patients suffering from bronchial asthma (BA) with associated gastroesophageal reflux disease (GERD). 101 patients with BA and associated GERD being under study were divided into three groups depending on the way of application of the anti-reflux therapy. Patients from the I group were administered Omeprazole--20 mg per day, patients from the II group received Omeprazole--mg per day, and patients from the III group were administered Esomeprazole--40 mg per day, the treatment courses of each group amounting to eight weeks. The clinical efficiency indices of pulmonary (CEIP) and esophageal (CEIE) symptoms were calculated when the results of the study were summed. The decrease of CEIP and CEIE against the background of the anti-reflux therapy (for example, the CEIP of patients from the III group decreased from 14.9 +/- 1.5 (before the treatment) to 2.9 +/- 0.6 points following eight weeks of treatment (p < 0.001), and the CEIE decreased from 9.0 +/- 1.2 (before the treatment) to 1.7 +/- 0.3 points following eight weeks of treatment (p < 0.001)) has demonstrated the reliable improvement of the clinical course of BA with associated GERD. We failed to discover any essential differences between the impact of Omeprazole in the dose equal to 40 mg per day and the impact of Esomeprazole in the same dose on the CEIP. Thus, the CEIP amounted to 10.1 +/- 1.2 points in the II group and 8.5 +/- 1.4 points in the III group (p > 0.05) after the fourth week of treatment. At the same time, we have discovered a reliable advantage of Esomeprazole as compared with Omeprazole for the improvement of the CEIE. The therapy of BA with associated GERD with Omeprazole in the dose equal to 40 mg per day or Esomeprazole in the dose equal to 40 mg per day contributes to the reliable improvement of both pulmonary and esophageal symptoms, and application of Esomeprazole results in a faster reduction of bronchial obstruction and gastroesophageal reflux. PMID- 14556545 TI - [Metabolic syndrome and the pancreas. Status of blood circulation in the pancreas in metabolic syndrome in patients with different types of hyperlipoproteinemia ]. AB - A study of blood circulation in the pancreas of 42 patients with the metabolic syndrome by the method of intracavitary rheography revealed a drop in the intensity of terminal blood circulation and presence of venous congestion with the activation of venous-arterial shunting. Hemodynamic disorders directly depended on the adiposity degree and patients' age, being evident as signs of a more obvious disorder of terminal blood circulation in patients with HLP of the II and III type. PMID- 14556546 TI - [Morphofunctional organization of pancreatic secretory activity (a new paradigm)]. AB - This is a review of physiological, morphological and clinical studies of pancreas secretion with the emphasis placed on pancreatic enzymes. It was established that different parts of the duodenal mucous coat have different chemosensory thresholds to the stimulators and inhibitors of pancreatic secretion. Secretory regions of the pancreas having a modular organization are also functionally different. The module is a set of pancreatic acinar and ductal cells (a secretory component), a system of ducts with their valve structures and a microdepository of secretion (a transport component) with control mechanisms specific for the module. Pancreatic secretion is a result of stimulation and inhibition of specialized secretory and transport modules. PMID- 14556547 TI - [Basis for the use of a new galena-based substance from plants for prevention and treatment of cholelithic disease]. AB - With the purpose of prevention and treatment of cholelithic disease, a new galena based phytocomposition having the choleritic, cholekinetic, spasmolytic and hypocholesterolemic effects at the same time was administered to patients along with lithotripsy. Clinical manifestations of cholecystitis disappeared after a daily intake of the phytocomposition during 30 days in 96 patients out of 418 people having the diagnosis of cholelithic disease and high initial concentrations of?-lipoproteins and cholesterol in the blood serum. The level of lipids dropped reliably, and it became much easier to use the lithotripsy method. Due to the decrease of bile viscosity, it was possible to note easier elimination of fragments from the gallbladder. During the monotherapy with the galena-based phytocomposition, full litholisis of single (up to 30 mm) or multiple (up to 50% of the gallbladder volume) calculi was determined in seven patients out of 47 (17%) during the period from two to six months. PMID- 14556548 TI - [Indices of cholecystokinin secretion, vegetative regulation of heart rate and anxiety level in patients with a motor dysfunction of the gallbladder]. AB - We examined 36 patients with biliary diseases (14 men and 22 women) at the age of 19-62 (19 patients with chronic non-calculous cholecystitis, 11 patients with cholelithiasis, and 6 patients with gallbladder dysfunctions) as well as 14 practically healthy people from the control group. Methods of the study included dynamic sorbite echography of the gallbladder, determination of serum cholecystokinin by the radioimmunoassay (RIA) as well as examination of variability of the cardiac rate and anxiety level. Patients with a motor dysfunction of the gallbladder suffer from disorders of the correlation between cholecystokinin secretion, the functional state of the vegetative nervous system and the nature of the motor and evacuation function of the gallbladder. PMID- 14556549 TI - [Endothelial dysfunction in patients with chronic colitis]. AB - The work shows that patients with chronic colitis have an expressed endothelial dysfunction, which becomes apparent as an increase of the level of vasoconstrictor endothelin-1 and decrease of the level of vasodilator nitric oxide. The obtained data indicate the role of the endothelial dysfunction in the pathogenesis of chronic colitis. The efficiency of the complex therapy with the use of folic acid and zinkteral for the restoration of the endothelium function is marked in comparison with the group of the patients treated with the help of the basic therapy. PMID- 14556550 TI - [Combination therapy of chronic pancreatitis (double-blind, placebo-controlled, pilot studies)]. PMID- 14556551 TI - [Hepon, promoter of local immunity in the complex therapy of dysfunctional microflora in bowel disorders]. AB - The promoter of local immunity Heponum contributes to the restoration of eubiosis and normalization of showings of the immune status in patients with post infection IBS. It is recommended to include Heponum in the complex therapy of chronic bowels diseases with the purpose of the restoration of normal microbiocenosis. PMID- 14556552 TI - [Bowel dysfunctional microflora: clinical significance and prospects for its therapy]. AB - Due to the application of complex drugs as well as synergistic action of drugs with a different mechanism of their activity, it is quite possible to restore the eubiosis of bowels. The up-to-date correction of biocenosis is an essential principle of the microecological approach to the maintenance of health in individuals and the entire population. PMID- 14556553 TI - [Adequate correction of gastric functional structure in surgery of peptic ulcer]. AB - Self-regulating mechanisms of homeostasis of the digestive apparatus are interrelated and interdependent with the algorithm of alimentary chymus movement from stomach into bowels. As the natural digestive tract is disturbed (pylorectomy, shutting-off the duodenum and drainage of stomach), the pathology leaves the frameworks of one organ and takes on a polyorgan character. A new principle of operation--adequate correction of functional structures of stomach ("repair" of the organ)--is suggested. It is based on the original plastic surgery of the pyloric canal, which enables us to maintain the natural algorithm of the gastroduodenal "passage", ulcerectomy and correction of acid formation. The status of regulating systems was estimated by the vital activity of the mucous membrane of the operated stomach, the trophism of which depends on the total influence of "feedbacks", as has been known. PMID- 14556554 TI - [Pancreatic exocrine secretion in experimental acute postoperative pancreatitis]. AB - Transformations of parameters of pancreatic exocrine secretion after the direct operations on the pancreas of a dog were studied during the experiment. It was determined that the early post-operative period proceeds in the conditions of the expressed pancreatic hypersecretion, hyperenzymemia and ductal hypertension. Their intensity reduces as the destructive process in pancreas develops. This results in changes of pancreatic exosecretion in two phases: the increasing and decreasing hypersecretion. The role of augmentation of the secretory activity of pancreas in pathogenesis of the acute postoperative pancreatitis is discussed. Its development was observed in all operated animals. This enabled us to resolve all contradictions and to prove the expediency of the application of pancreas secretion inhibitors at the prevention of acute post-operative pancreatitis. PMID- 14556555 TI - [Morphological study of bile in the diagnosis of biliary diseases]. AB - The purpose of this work was to study principles of bile structurization in healthy people and patients with various biliary diseases. 160 patients with different biliary diseases and other diseases of the hepatopancreaticoduodenal zone were examined. Samples of gallbladder bile were taken from corpses of young men, who did not have any diseases of the hepatopancreaticoduodenal zone. Their diagnoses were confirmed by an ultrasound morphological study. Bile was studied by the cuneate dehydration and viscosimetric methods. The structure of facies of gallbladder bile under conditions of absence of diseases of the hepatopancreaticoduodenal zone was shown. The facies have a wide convex peripheral zone (a cushion) without any well-defined border. The central part of the facies is amorphous or fine-grained, sometimes with occasional inclusions of larger crystals. Markers of exacerbation of chronic cholecystitis (dendrites in the central zone of bile facies) as well as signs of the presence of a malignant neoplasm touching the bile (large diamond-shaped crystals in the central zone of bile facies where dendrites begin) were discovered. PMID- 14556557 TI - Get physicians on your side to multiply DM results. PMID- 14556556 TI - [Clinical assessment of the efficacy of early enteral nutrition with the help of "peptamen" nutriment in patients who underwent gastrectomy due to stomach cancer]. AB - The protein-energetic insufficiency in cancer patients aggravates the course of the disease and leads to various complications in the post-operative period including lethal outcomes. The purpose of the study is to assess the efficiency of the use of the Peptamen compound in the early post-operative period in patients with moderate and serious degrees of protein-energetic insufficiency, who underwent gastrectomy due to stomach cancer. Two groups of patients were studied differing by the method of nutritional support. The use of the Peptamen compound in the early post-operative period leads to the improvement of the absorption of nutrients, increase in the functional activity of the intestine and immunomodulation, and as a result of this, to the reduction of post-operative complications and fast restoration of homeostasis. PMID- 14556558 TI - Lay educators shore up diabetes DM efforts in Hispanic communities. AB - Diabetes impacts Hispanics at twice the rate of Caucasians, and traditional DM efforts are not adequately reaching this fast-growing minority group. However, a novel approach that makes use of 'lay educators' is showing promise at a number of sites across the country. And developers suggest that the model may have potential use in other minority groups as well. PMID- 14556559 TI - Modest approach toward diabetes DM can deliver big returns. AB - One DM effort that includes a maximum of two outbound calls to diabetic participants has had considerable success in bringing physician-prescribed treatment in line with recommended guidelines, boosting self-care skills, and cutting costs. Find out how this pharmacist-driven effort is getting results. PMID- 14556560 TI - New inner-city asthma interventions put social workers in the driver's seat. AB - Why? Because research shows that, unlike so many failed strategies, a social worker-based intervention can boost outcomes and cut costs. The critical element seems to be the social worker's ability to remove barriers that commonly sabotage other efforts. See how this alternative approach to asthma DM is being implemented. PMID- 14556562 TI - Hyperoxia may reduce energetic efficiency in the trained rat. AB - BACKGROUND: Several studies have been conducted in recent years in the attempt to improve running performance by the use of hyperbaric oxygen, but there is disagreement as to whether this has any beneficial effect. The purpose of this study was to measure the effect of 24 h breathing 100% O2 in normobaric conditions on energetic efficiency in the trained rat. METHODS: Experiments were carried out on trained rats whose oxygen consumption was evaluated during the training period and on its completion. At the end of the training period, the rats were divided into two groups: 1) rats exposed to air (21% O2) in normobaric conditions; and 2) rats exposed to 100% O2 in normobaric conditions. In addition, two groups of sedentary rats were used: 3) sedentary rats exposed to air (21% O2) in normobaric conditions; and 4) sedentary rats exposed to 100% O2 in normobaric conditions. Energetic efficiency was estimated by measuring O2 consumption at submaximal exercise (45 m.min-1, 10 degrees incline). RESULTS: Training alone reduced O2 consumption by 18% during submaximal exercise. Exposure to 100% oxygen for 24 h in normobaric conditions reversed the effect of complete training by elevating the O2 consumption by 17%, which was close to the oxygen consumption of the rats during the incomplete training period. CONCLUSIONS: Our results suggest that prolonged exposure to hyperoxia induces a reduction in the energetic efficiency of the trained rat. The relevance of these findings to sports and diving is discussed. PMID- 14556561 TI - Acceleration-induced near-loss of consciousness: the "A-LOC" syndrome. AB - BACKGROUND: There is an insidious phenomenon that can occur when aircrew are exposed to +Gz stress even at levels that are insufficient to cause +Gz-induced loss of consciousness (G-LOC). Under these circumstances aircrew exhibit an altered state of awareness that was termed Almost Loss of Consciousness (A-LOC) by the U.S. Navy in the late 1980's. A-LOC is a syndrome that includes a wide variety of cognitive, physical, emotional, and physiological symptoms. While A LOC has been observed in centrifuge studies and reported in flight for over 15 yr, a definitive description of the syndrome does not exist. METHODS: Nine subjects were exposed to short +6, 8, and 10 Gz pulses of increasing duration until they experienced G-LOC. Instrumentation included two channels of ECG and near infrared spectroscopy (NIRS) to measure relative cerebral tissue oxygenation (rSo2). Subjects indicated +Gz-induced visual symptoms (light loss, LL) by pressing a switch when LL began and releasing it when total vision was restored. Short-term memory loss was assessed using a simple math task. Data analysis included a description and the time course of the physical, physiological, cognitive, and emotional responses. RESULTS: There were 66 episodes of A-LOC that were identified out of a total of 161 +Gz pulse exposures. Many incidents of sensory abnormalities, amnesia, confusion, euphoria, difficulty in forming words, and reduced auditory acuity were documented. Often these responses occurred in multiple subjects and at different +Gz levels. One of the most common symptoms was a disconnection between cognition and the ability to act on it. There was a significant reduction in rSo2 over baseline, greater overshoot in rSo2 (increase in oxygenation above baseline after the +Gz exposure), faster fall in rSo2 during +Gz stress, and prolonged recovery time associated with A-LOC as compared with +Gz exposures without symptoms. CONCLUSION: Evaluation of the range of symptoms associated with A-LOC can lead to a program to increase pilots' awareness of the phenomenon and further our understanding of the relationship between the outward symptoms and the underlying physiological changes. PMID- 14556563 TI - Negative pressure breathing enhances nitrogen elimination. AB - BACKGROUND: The rate of nitrogen elimination during decompression is primarily dependent on tissue perfusion and, therefore, cardiac output (CO). Negative pressure breathing (NPB) is thought to increase CO by enhancing venous return and, consequently, the rate of nitrogen elimination. This notion was investigated in the present study. METHODS: Nitrogen elimination was measured in five subjects lying supine while breathing a nitrogen free gas (79% Ar, 20% O2) supplied at -15 cm H2O (-1.5 kPa) or at atmospheric pressure. RESULTS: Over a 2-h washout period, NPB increased nitrogen elimination by 39.2 +/- 21.7% (mean +/- SD), increased calf blood flow by 34.1 +/- 34.6%, and elevated BP slightly during the last half of the experiment. Negative pressure breathing did not significantly change CO; the increase in nitrogen elimination may have been due to redistribution of blood flow. CONCLUSIONS: Negative pressure breathing appears to be a useful means of increasing nitrogen elimination and should be considered in situations such as decompression or treatment of decompression sickness where this effect may be beneficial. PMID- 14556564 TI - Submaximal exercise with self-contained breathing apparatus: the effects of hyperoxia and inspired gas density. AB - BACKGROUND: The self-contained breathing apparatus (SCBA) used by firefighters, and other working in dangerous environments, adds an external resistance to expiration, which increases expiratory work during heavy exercise. HYPOTHESIS: Compressed air is typically used with the SCBA and we hypothesized that changing the inspired oxygen concentration and/or gas density with helium would reduce the external expiratory resistance. METHODS: On separate days, 15 men completed four 30-min bouts of treadmill exercise dressed in protective clothing and breathing the test gases through the SCBA. Four different gas mixtures were assigned in random order: [compressed air (NOX: 21% O2, 79% N2), hyperoxia (HOX: 40% O2, 60% N2), normoxic-helium (HE-OX: 21% O2, 79% He), and helium-hyperoxia (HE-HOX: 40% O2, 60% He)]. RESULTS: Compared with NOX, the two helium mixtures (but not HOX), decreased the external breathing resistance and all three gas mixtures decreased the peak expired mask pressure and the ventilatory mass moved. Both hyperoxic mixtures decreased blood lactate and the rating of perceived exertion was decreased at 30 min with HE-HOX. CONCLUSIONS: These results demonstrate that the helium-based gas mixtures, and to a lesser extent HOX, reduce the expiratory work associated with the SCBA during strenuous exercise. PMID- 14556565 TI - Contrast sensitivity after +Gz acceleration. AB - BACKGROUND: This study was designed to determine the extent and duration of contrast sensitivity (CS) loss after high sustained +Gz acceleration in a centrifuge. METHODS: The subjects were 12 healthy male flight surgeons between 20 and 22 (mean = 21.1) yr of age. The human centrifuge at the Aviation Physiology Research Laboratory in Tainan, Taiwan, was used to expose the subjects to an acceleration profile. Each subject experienced three centrifuge runs made up of one gradual onset and two rapid onset profiles. Contrast sensitivity (CS) was measured before, and at 5 min, 10 min, and 20 min for the right eye; and 7 min, 12 min, and 22 min for the left eye after the acceleration. Both eyes were measured with the right eye being tested first. RESULTS: There was a generalized depression of CS at 5-12 min for both eyes. The depression was more severe at low and medium frequencies (1.5, 3.0, 6.0 cycles per degree, cpd) than at a high spatial frequency (18.0 cpd). There was a significant decrease in CS of the right eye at 1.5 cpd (p < 0.05 between control and 5 min), and on the left eye at 3.0 cpd (p < 0.05 between control and 7 min, p < 0.05 between control and 12 min) and 6.0 cpd (p < 0.05 between control and 12 min). The CS loss was more obvious at 5 12 min for both eyes, and there was only partial recovery at 22 min after the acceleration. CONCLUSIONS: These results demonstrated that +Gz acceleration is associated with CS loss. The recovery time was greater than expected. Factors other than ocular blood flow may be involved in the prolonged CS loss. PMID- 14556566 TI - The risk of altitude decompression sickness at 12,000 m and the effect of ascent rate. AB - INTRODUCTION: Loss of aircraft cabin pressurization can result in very rapid decompression rates. The literature contains reports of increased or unchanged levels of altitude decompression sickness (DCS) resulting from increasing the rate of decompression. We conducted two prospective exposure profiles to quantify the DCS risk at 12,192 m (40,000 ft), and to determine if there was a greater DCS hazard associated with a much higher rate of decompression than typically used during past DCS studies. METHODS: The 63 human subjects participated in 80 altitude chamber decompression exposures to a simulated altitude of 12,192 m (2.72 psia; 18.75 kPa) for 90 min, following preoxygenation with 100% oxygen for 90 min. Half of the subject-exposures involved an 8-min decompression (1,524 mpm; 5,000 fpm) and the other half experienced a 30-s decompression (mean of 24,384 mpm; 80,000 fpm). Throughout each ascent and exposure, subjects were seated at rest and breathed 100% oxygen. At altitude, they were monitored for precordial venous gas emboli (VGE) and DCS symptoms. RESULTS: The higher decompression rate yielded 55.0% DCS and 72.5% VGE and the lower rate produced 47.5% DCS and 65.0% VGE. Chi square and log rank tests based on the Kaplan-Meier analyses indicated no difference in the incidence or onset rate of DCS or VGE observed during the two profiles. CONCLUSION: Decompression rate to altitude up to 24,384 mpm was found not to have an effect on DCS risk at altitude. However, research is needed to define the DCS risk with decompression rates greater than 24,384 mpm. It was also found that the onset time to DCS symptoms decreases as altitude increases. PMID- 14556568 TI - Case-control study of risk factors for fatal and non-fatal injury in crashes of civil aircraft. AB - BACKGROUND: There have been few studies of the risk factors for fatal injury in air crashes, and none of risk factors for all serious injury (fatal and non fatal). OBJECTIVE: To identify the potentially modifiable risk factors for fatal and non-fatal injury in civil aircraft crashes in New Zealand. METHODS: We analyzed records from all reported civil aircraft crashes in New Zealand (1988 1994). Air crash data from the official databases were merged with nationwide injury records and information obtained from coroners' files. Pilots-in-command who were fatally injured were compared with pilots-in-command who were not fatally injured using 50 variables covering pilot, aircraft, environmental, and operational characteristics. A second analysis compared pilots-in-command who were seriously injured (either fatally or non-fatally) with those who were involved in a crash but not hospitalized with an injury. Univariate and multivariate logistic regression analyses were then conducted to estimate the odds associated with each of the factors identified by the above comparisons. RESULTS: The most significant risk factors for fatal and serious injury were aerobatic flight, post-crash fire, not having a certificate of airworthiness, and off-airport location. Environmental characteristics including terrain type, wind, rain, and elevation of the crash site were identified as risk factors, as was being under instruction. Flying a twin-engine aircraft was a risk factor for fatal injury, while piloting a microlight aircraft was a risk factor for all serious injury. CONCLUSION: Environmental and operational factors, rather than pilot or aircraft characteristics, are the key determinants of the injury outcome of civil aircraft crashes. PMID- 14556567 TI - Association between right-to-left shunts and brain lesions in sport divers. AB - BACKGROUND: Recent studies suggest that healthy sport divers may develop clinically silent brain damage, based on the association between a finding of multiple brain lesions on MRI and the presence of right-to-left shunt, a pathway for venous gas bubbles to enter the arterial system. METHODS: We performed echocontrast transcranial Doppler sonography in 42 sport divers to determine the presence of a right-to-left shunt. Cranial MRI was carried out using a 1.5 T magnet. A lesion was counted if it was hyperintense on both T2-weighted and T2 weighted fluid attenuated inversion recovery sequences. To test the hypothesis that the occurrence of postdive arterial gas emboli is related to brain lesions on MRI, we measured postdive intravascular bubbles in a subset of 15 divers 30 min after open water scuba dives. RESULTS: Echocontrast transcranial Doppler sonography revealed a right-to-left shunt in 16 of the divers (38%). Only one hyperintensive lesion of the central white matter was found and that was in a diver with no evidence of a right-to-left shunt. Postdive arterial gas emboli were detected in 3 out of 15 divers; they had a right-to-left shunt, but no pathologic findings on cranial magnetic resonance imaging. CONCLUSIONS: Our data support the theory that right-to-left shunts can serve as a pathway for venous gas bubbles into the arterial circulation. However, we could not confirm an association between brain lesions and the presence of a right-to-left shunt in sport divers. PMID- 14556569 TI - Peptic ulcer data from 4 years of endoscopy in 955 pilots of the Japan Air Self Defense Force. AB - BACKGROUND: The treatment of peptic ulcer disease has undergone profound changes due to the recognition of Helicobacter pylori as a causative factor. A survey of medical records was made to determine the prevalence of peptic ulcer among pilots of the Japan Air Self Defense Force (JASDF) and to decide on a possible change in JASDF medical policy toward an ulcer-treatment regime involving therapy to eradicate Helicobacter pylori. METHODS: The subjects were 955 male pilots, age 40 or older. Between 1996 and 1999, they underwent gastrointestinal endoscopy 2.47 times on average. Annual ulcer rates and recurrence rates were obtained from the endoscopic file data. Smoking habits and use of NSAIDs were also assessed as important risk factors for peptic ulcer. RESULTS: The detection rate of open ulcer for each year was 2.3-3.1% in the stomach and 1.9-4.4% in the duodenum. For ulcers including scarring, the corresponding figures were 7.3-9.5% and 12.7 19.9%. The recurrence rate from S1 scars in the duodenum was 34%, significantly higher than that from S2 scars (7%) (p < 0.0005). There was no significant difference in recurrence rate between S1 scars and S2 scars in the stomach. There was a significant association between gastric ulcer and smoking (p < 0.0005). None of the pilots took long-term NSAID medications. CONCLUSIONS: Peptic ulcer occurs more frequently in the JASDF pilots than in the general population. S1 scarring in the duodenum, as well as open ulcers in either stomach or duodenum, are candidates for Helicobacter pylori eradication therapy if the bacterium is detected. Other types of scars are also candidates for this therapy when pilots have symptoms related to infection with Helicobacter pylori. PMID- 14556571 TI - Inflight workload assessment: comparison of subjective and physiological measurements. AB - INTRODUCTION: Assessment of pilot workload during flight is an important aviation safety consideration. The aim of this study was to assess inflight pilot workload using both physiological and multidimensional subjective-ratings measurements (heart rate and NASA Task Load Index, respectively), comparing relative sensitivity during the four phases of flight: take-off, cruise, approach, and landing. METHODS: Ten male pilots volunteered to participate in the trials, which took place in a Boeing 747-400 flight simulator. Electrocardiography was performed throughout the test using the portable Cardiovis ECG system. Mean heart rate (HR) and incremental heart rate (delta HR) were considered indices of physiological workload. RESULTS: Peak HR was observed during take-off (83.2 bpm) and landing (88.6 bpm); moreover, delta HR was also greatest (14.2 bpm and 18.8 bpm). The Task Load Index (TLX) scale revealed that mental and performance demands were essential components of workload during flight. In addition, temporal demand was an important component of workload during take-off and physical demand was significant during cruise. Analysis of correlation revealed that the delta HR is significantly related to TLX scores (r = 0.81, n = 40). CONCLUSIONS: Management of the individual sources of stress, which tend to become predominant during different flight phases, should be emphasized in periodic recurrent training. For example, a pilot must be trained to cope with the increased temporal stresses associated with take-off. In addition, the recommendations will be concerned with maintaining vigilance, task allocation between pilots, and inflight rest during long-haul cruise. PMID- 14556570 TI - Perceived fatigue for short- and long-haul flights: a survey of 739 airline pilots. AB - BACKGROUND: Fatigue-related incidents in aviation may be self-reported by pilots in confidential systems. The aim of this study was to clarify what fatigue means to pilots on short- and long-haul flights (SHF and LHF, respectively). METHODS: Questionnaires were distributed to pilots through four airlines. Questions concerned the perceived causes of fatigue, its signs and symptoms in the reporting pilot and observed in others, as well as the strategies used to minimize its impact. RESULTS: Of 3,436 questionnaires distributed, 739 (21.5%) were returned. For LHF, fatigue was seen as mainly due to night flights (59%) and jet lag (45%). For SHF, fatigue was caused by prolonged duty periods (multi segment flights over a sequence of 4 to 5 d) (53%) and successive early wake-ups (41%). Self-reported manifestations of fatigue in 60% of LHF pilots and 49% of SHF pilots included reduction in alertness and attention, and a lack of concentration. Signs observed in other crewmembers included an increase in response times and small mistakes (calculation, interpretation). When pilots were tired, all the flying tasks seemed to be more difficult than usual. In both LHF and SHF, rest and sleep management were the primary strategies used to cope with fatigue. Analysis showed that duty time is a major predictor of fatigue, but that it cannot be considered independently from the other contributory factors. CONCLUSION: For both LHF and SHF, pilots reported acute fatigue related to sleep deprivation, due mainly to work schedules: night flights, jet-lag, and successive early wake-ups. These causal factors could easily be assessed in investigation of accidents and incidents. PMID- 14556572 TI - Acute soft tissue neck injury from unexpected acceleration. AB - Recent evidence has identified a risk of neck injury resulting from exposure to high sustained G force during operation of fast jet military aircraft. This review considers the risk of acute soft tissue neck injury when exposure to acceleration is unexpected by the individual. In a military jet, this might occur to a back seat occupant such as an instructor or weapons operator/navigator, but unexpected forces may also be encountered by roller coaster riders and human centrifuge users under certain conditions. The mechanisms of injury in this situation are examined, the effect of acceleration axes (Gx, Gy, and Gz) are considered, and relevant experimental data are evaluated. These data are used to determine what influence the unexpected nature of the acceleration exposure has on the risk of neck injury. Injury below previously established 'tolerance' limits is possible, with some individuals being at risk from unexpected acceleration exposure above around +2 to . PMID- 14556574 TI - Hand discomfort following heliox chamber dives. AB - During a series of dry chamber dives using compressed heliox, five attendants and one wet diver experienced eight episodes of hand discomfort, the character of which was atypical of limb pain during decompression sickness. Although immersed for most of the dive, during the compression and decompression phases, the wet diver's hands were out of the water and hence exposed to the helium-containing chamber atmosphere. In all cases, symptoms resolved within a maximum of 48 h. There was no response to hyperbaric oxygen therapy in the three cases that presented before spontaneous resolution. While the attendants wore dry suits to minimize skin absorption of helium, their hands, were exposed to the heliox atmosphere. After the first six cases of hand symptoms, a dry glove assembly was added to prevent helium absorption through the exposed hand. Two cases of hand discomfort occurred following the addition of the dry glove assembly to the dry suit. In both cases, the symptoms were less severe and resolved over a significantly shorter time period. Adoption of the dry gloves resulted in the incidence of hand discomfort among attendants falling from 25% (5/20) to 2.4% (2/84) (p = 0.005). Possible mechanisms of causation of this hand discomfort, thought to be the result of local tissue absorption of helium, are discussed. PMID- 14556573 TI - Diabetes mellitus type 2 in aviators: a preventable disease. AB - INTRODUCTION: The current epidemic of obesity and resultant diabetes mellitus type 2 (DMT2) is a tsunami that will impact healthcare worldwide and lap over into aerospace medicine. Metabolic syndrome (MBS) is the major link between obesity and DMT2. METHODS: A review of U.S. Air Force Aeromedical Consult Service (ACS) records was accomplished looking at aviators with a diagnosis of DMT2. Case reports of three flyers with DMT2 are presented and discussed. Other aeromedical agencies were contacted regarding their experiences and this information was summarized. A literature review on DMT2, obesity, and metabolic syndrome was accomplished. RESULTS: Of 70 charts for flyers identified with diabetes mellitus at the ACS between 1975 and 2000, over 95% were for DMT2. The mean body mass index for these aviators was 26.2. Currently, all services grant restricted waivers for some aviators with DMT2, none in high performance, single-seat aircraft. The FAA is currently allowing most flyers with stable DMT2 to operate aircraft in all categories with specific restrictions. DISCUSSION: Obesity and metabolic syndrome are becoming increasingly prevalent in the aviation community. Aggressive actions to limit weight gain and identify those at risk for developing DMT2 must be considered for all populations. PMID- 14556575 TI - Cerebral infarction in a 24-year-old pilot. AB - Ischemic stroke is a rare event in young adults. We report on a 24-yr-old pilot with cerebral infarction of undetermined etiology, temporally associated with chain smoking. The patient exhibited dysphasia, stupor (confused consciousness), and right facial-nerve palsy. Computed-tomography revealed a low-density area in the left insular cortex. Cerebroangiography showed severe stenosis in a branch of the left middle cerebral artery. After admission, the patient made a rapid and uneventful recovery within 72 h. MRI showed an area of hyperintensity on T2 weighted images 2 mo after the attack. Based on the hyperintense area on FLAIR (fluid attenuated inversion recovery sequence) images obtained in MRI performed 10 mo after the attack, we diagnosed a cerebral infarction. In the Japan Air Self Defense Force, cerebral infarction is an aeromedically disqualifying condition. However, in the evaluation 2 mo after the attack, differentiation from reversible ischemic neurological deficit was difficult. We discuss the criteria used for diagnosis and the risk factors for cerebral infarction in young adults, as well as the aeromedical disposition of young pilots. PMID- 14556576 TI - [Genetics and arterial hypertension: 3 approaches to decode a complex disease]. AB - Human arterial hypertension is a complex trait, partly determined by genetic factors. From the analysis of familial studies, it has been estimated that approximately 30% of the blood pressure variance within a population was of genetic origin. Three main types of human studies have been undertaken to try to identify susceptibility genes to hypertension. The first one corresponds to the systematic analysis of the so-called candidate genes, i.e. genes encoding proteins, enzymes, receptors, which are known to belong to pathways controlling blood pressure. Up to now, the most interesting results have been obtained on genes encoding the renin angiotensin system, the a adducin, the G protein subunit beta 3, and adrenergic receptors. The genome wide scan approach corresponds to a systematic analysis of evenly spaced markers throughout the genome in sibling pairs or in more complex families. This second strategy has shown that there was not a single locus that was regularly found by several studies, but rather several possible loci which most often have not been replicated from one study to another one. Among those, the long arm of the human chromosome 17 (17q12-q21) is in synteny with a blood pressure locus found in spontaneously hypertensive rats. The third approach, up to now the most successful, corresponds to the identification of major genes involved in rare Mendelian forms of hypertension. For example, genes responsible for Liddle syndrome, glucocorticoid remediable aldosteronism, apparent mineralocorticoid excess have been characterized and have demonstrated the importance of sodium and water homeostasis in blood pressure control. PMID- 14556577 TI - [Recent development of recommendations for the screening and treatment of arterial hypertension]. AB - Hypertension is a very prevalent condition which alone, or in association with various risk factors including smoking, hypercholesterolemia and diabetes mellitus, may lead to fatal or non-fatal stroke or myocardial infarction. Antihypertensive treatment is effective at preventing cardiovascular events. However, only a minority of patients with hypertension ever experience a stroke or myocardial infarction and a few patients with well-controlled hypertension may suffer such events. Before committing a patient to lifelong treatment, the physician should ensure that high blood pressure is not a symptom of an underlying correctable disease, that hypertension is persistent and requires medication, and that prevention measures take into account all correctable risk factors. PMID- 14556578 TI - [Evaluation of the effect of magnetic fields on the secretion of melatonin in humans and rats. Circadian study]. AB - The consequences of electromagnetic exposure on human health are receiving increasing scientific attention and have become the subject of a vigorous public debate. In the present study we evaluated the effects of magnetic field on pineal function in man and rat. Two groups of Wistar male rats were exposed to 50-Hz magnetic fields of either 1, 10 or 100 microT. The first group was exposed for 12 hours and the second for 30 days (18 hours per day). Short-term exposure depressed both pineal NAT activity and nocturnal serum melatonin concentration but only with the highest intensity used (100 microT). Long-term exposure to a magnetic field significantly depressed the nighttime peak of serum melatonin concentration and pineal NAT activity with 10 and 100 microT. Our results show that sinusoidal magnetic fields altered the production of melatonin through an inhibition of pineal NAT activity. Both duration and intensity of exposure played an important role in this effect. In the second step of this study, thirty-two young men (20-30 years old) were divided into two groups (control group, i.e., sham-exposed: 16 subjects; exposed group: 16 subjects). The subjects were exposed to the magnetic field from 23 h to 08 h (i.e. for 9 h) while lying down. In one experiment the exposure was continuous, in the second one, the magnetic field was intermittent. No significant differences were observed between sham-exposed (control) and exposed men for serum melatonin and 6-sulfatoxymelatonin. In our last and more recent study, we looked for the circadian rhythm of melatonin in 15 men exposed chronically and daily for a period of 1-20 years, in the workplace and at home, to a 50 Hz (exposure 0.1 to > 0.3 microT) magnetic field. The results are compared to those for 15 unexposed men who served as controls. Blood samples were taken hourly from 2000 to 0800. Nighttime urine was also collected and analyzed. This work shows that subjects exposed over a long period (up to 20 years) and on a daily basis to magnetic fields experienced no changes in their plasma melatonin level, their urinary 6-sulfatoxymelatonin level, or the circadian rhythm of melatonin. It thus clearly rebuts the "melatonin hypothesis" that a decrease in plasma melatonin concentration--or a disruption in its secretion--explains the occurrence of, clinical disorders or cancers possibly related to magnetic fields. PMID- 14556579 TI - [Sino-nasal polyposis. Evaluation of the efficacy of combined local and general corticotherapy in a series of 100 consecutive patients with a 3-year follow-up]. AB - Nasal polyposis accounts for 40% of chronic nasal disease. The part played by surgery seems to be steadily growing if the number of publications dedicated to this approach is any yardstick. The medical treatment remains however the undisputed therapeutic mainstay but trials dedicated to the assessment of its overall efficacy are rather scarce. The aim of this study is focused on the evaluation of a dual modality, topical and systemic, over a follow-up period of three years. A total of 100 subjects were treated according to a standardized therapeutic protocol combining a short-term oral administration of a corticosteroid (prednisolone) and a daily intranasal spray of an other steroid (beclomethasone). Over the follow-up period of three years, this dual modality proved to be successful in 85% of the subjects; only 15% had to undergo surgery after its failure. The average symptom reduction reached an improvement rate varying from 58 to 80%, according to the symptom type. The daily dosage of prednisolone and beclomethasone was progressively decreased while the gain in nasal comfort was being preserved. Management of nasal polyps should be primarily medical. Resorting to surgical procedures should not be envisaged before a trial of dual steroid therapy under strict compliance to treatment. The single character of this long follow-up and the importance of the series of patients included in the protocol make all the originality of this work which shows for the first time the long-term improvement after medical treatment. PMID- 14556580 TI - Sharing your work: building knowledge about nursing care quality. PMID- 14556581 TI - Process improvement for asthma. An integrated approach. AB - At least 18 million people in the United States have asthma. Despite the publication of national guidelines, the delivery of care has not substantially changed. This article describes a program at Chicago city sites to improve the delivery of care to adults and children with asthma. Using consistent comprehensive patient education materials, innovative provider education, and a variety of continuous quality improvement interventions including creation of a designated practitioner, this project enhances partnership between patients and health care providers. PMID- 14556583 TI - Patient satisfaction of emergency nursing care in the United States, Slovenia, and Australia. AB - Patient satisfaction with nursing care in the emergency department (ED) was measured in 616 patients in California and Kentucky in the United States, Slovenia, and rural and urban Victoria in Australia. Patients were almost equally divided between men and women, primarily white, with a mean age of 44. They responded to the caring and teaching subscales of the Consumer Emergency Care Satisfaction Scale (CECSS). While scores indicated no differences in caring, the US and Slovenian groups reported greater satisfaction with teaching than did the Australian group. The subscale alpha coefficients were .87 for the caring subscale and .92 for the teaching subscale. PMID- 14556582 TI - Process-based costing. AB - Understanding how quality improvement affects costs is important. Unfortunately, low-cost, reliable ways of measuring direct costs are scarce. This article builds on the principles of process improvement to develop a costing strategy that meets both criteria. Process-based costing has 4 steps: developing a flowchart, estimating resource use, valuing resources, and calculating direct costs. To illustrate the technique, this article uses it to cost the care planning process in 3 long-term care facilities. We conclude that process-based costing is easy to implement; generates reliable, valid data; and allows nursing managers to assess the costs of new or modified processes. PMID- 14556584 TI - Perceptions of utilization review nurses. "Nurses like us". AB - The purpose of this research was to describe the characteristics of nurses who perform telephonic care quality assessment, or utilization review, prior to, concurrent with, and after medical interventions and hospitalizations. It compares these nurses demographically to nurses employed in other settings, discusses the nurses' perceptions of the nature of their work, examines assertions that utilization review is a nursing activity, and documents the nurses' education and training for ethical decision-making. Implications for nursing care quality are discussed. PMID- 14556585 TI - Functional status improvement in the elderly following coronary artery bypass graft. AB - This study assessed functional status changes among elderly persons (aged 65+) during the first 2 years following an elective coronary artery bypass graft. Physical functional status increased 39.1% from baseline to 1 year and 2.1% from 1 to 2 years. Role functioning increased 42.7% at 1 year and slightly decreased by 2.1% from 1 to 2 years postsurgery. From baseline to 1 year, female patients nearly doubled the percent gain of men for both Physical Function (.3% vs +33.9%, respectively) and Role Function (+62.0% vs +37.8%, respectively). Physical Function scores continued to increase from 1 to 2 years for women (+9.5%) compared to men (+0.7%). These findings suggest that elderly persons can expect postsurgical functional status to increase steadily during the first 2 years after surgery. PMID- 14556586 TI - School-based health centers. Statewide quality improvement program. AB - School-based health centers have been proposed as an avenue to provide health care for vulnerable youth. Colorado has 45 school-based health centers that provided 50,000 visits to 18,600 vulnerable youth in the 1998-1999 school year. Sustainability of the centers, however, depends on the ability to obtain an ongoing stream of revenue including managed care contracts, which in turn depend on the center's ability to meet the standard of care in the community. School based health centers should meet standards of care similar to those of community health centers including certification, credentialing of providers, and a systematic evaluation of the outcomes of services. This article reports on a statewide quality improvement program that established certification standards for school-based health centers, verified the credentials of providers at the centers, and measured quality of care in centers across the state. PMID- 14556587 TI - Quality health care in an era of limited resources. Challenges and opportunities. AB - Quality health care is a complex phenomenon. The factors contributing to quality in health care are as varied as the strategies needed to achieve this elusive goal. This article examines the impact of medical errors, health finance (insurance), moral hazard, and provider shortages on the quality of health care today in the United States. Emphasis is given to the need to restore public trust and to create a health care system that maximizes scarce physical, fiscal, and human resources. PMID- 14556588 TI - United States Census 2000 population with bridged race categories. AB - OBJECTIVES: The objectives of this report are to document the methods developed at the National Center for Health Statistics (NCHS) to bridge the Census 2000 multiple-race resident population to single-race categories and to describe the resulting bridged race resident population estimates. METHOD: Data from the pooled 1997-2000 National Health Interview Surveys (NHIS) were used to develop models for bridging the Census 2000 multiple-race population to single-race categories. The bridging models included demographic and contextual covariates, some at the person-level and some at the county-level. Allocation probabilities were obtained from the regression models and applied to the Census Bureau's April 1, 2000, Modified Race Data Summary File population counts to assign multiple race persons to single-race categories. RESULTS: Bridging has the most impact on the American Indian and Alaska Native (AIAN) and Asian or Pacific Islander (API) populations, a small impact on the Black population and a negligible impact on the White population. For the United States as a whole, the AIAN, API, Black, and White bridged population counts are 12.0, 5.0, 2.5, and 0.5 percent higher than the corresponding Census 2000 single-race counts. At the sub-national level, there is considerably more variation than observed at the national level. The bridged single-race population counts have been used to calculate birth and death rates produced by NCHS for 2000 and 2001 and to revise previously published rates for the 1990s, 2000, and 2001. The bridging methodology will be used to bridge postcensal population estimates for later years. The bridged population counts presented here and in subsequent years may be updated as additional data become available for use in the bridging process. PMID- 14556589 TI - The impact of the medical malpractice crisis on OB-GYNs and patients in southern New Jersey. AB - The results of this study indicate that the current medical malpractice crisis has created significant insurance affordability and accessibility problems for OB GYNs in southern New Jersey. Even more important is the result that shows this crisis as beginning to have a very real impact on patient access to care. The average cost for malpractice coverage per physician increased from $34,616.67 in 2000 to $78,818.18 in 2003. This represents a 128% increase in just four years. As dramatic as these premium increases appear, the fact that many physicians have had to reduce their coverage in order to afford any malpractice coverage at all has not yet been taken into account. The increase in premiums particularly affects solo practitioners, such as those practicing in the more rural areas of southern New Jersey, because they are unable to take advantage of the economies of scale that larger practices enjoy. It is also the case that premium increases are even higher for high-risk subspecialties, such as maternal and fetal medicine. Certified nurse midwives are just beginning to see significant increases in their premiums as well. Fourteen point three percent of practices got quotes from three carriers this year, and 12.2% of practices got more than three quotes. This may indicate that there are physicians who are unable to find affordable coverage. More than one quarter of OB-GYN practices (26.5%) reported that their current carrier dropped one or more of the physicians in their practice during the most recent renewal period. Close to half of the practices (49%) that responded to the survey indicated that they came across at least one carrier who refused to provide a quote for liability coverage, while 14% of the practices indicated that more than three carriers refused to provide a quote. This indicates a significant problem with access to coverage. The current malpractice crisis already has had an impact on the way in which physicians practice medicine. Most significant, thirteen practices in the region indicated that at least one of their physicians had stopped delivering babies, two additional practices indicated that they had ceased delivering or caring for high risk babies, and six practices indicated that at least one of their physicians had ceased performing surgical procedures. There are even more strategies that physicians in southern New Jersey are considering for the near future. Clearly, physicians believe that patient access to care is already a problem in southern New Jersey. If premiums do not stabilize in the very near future, there is every indication that southern New Jersey will have a severe access to care issue for its obstetrical patient population. Unfortunately, a recent report on the nation's medical malpractice insurance crisis predicts at least two more years of continuing financial problems for medical malpractice insurers. Despite the fact that New Jersey's state legislators ended their session without enacting any new tort reform laws, physicians across the state continue to apply pressure for passage of a bill to help the current medical liability crisis. Physicians plan a new round of lobbying and demonstrations beginning October 7, 2003. PMID- 14556590 TI - The HIPAA privacy rule and physician responses to medical-record requests in civil litigation. PMID- 14556591 TI - The editors of New Jersey medicine. PMID- 14556592 TI - Garbage and sewage disposal. 1906. PMID- 14556594 TI - Doctors as facilitators. PMID- 14556593 TI - Stroke. PMID- 14556595 TI - HIV-AIDS in 2003. PMID- 14556596 TI - HIV disease surveillance. Collaboration between medicine and public health. AB - Public Health Surveillance is critical to the management of programs designed to control the epidemic of HIV-AIDS. Surveillance defines changing trends, helps to formulate preventive initiatives and evaluate their effectiveness, and to allocate resources. Collaboration between clinical medicine and public health is essential to achieve reliable surveillance. PMID- 14556597 TI - Rapid diagnostic testing for HIV. Clinical implications. AB - There have been many advances in HIV counseling, testing, and referral over the past ten years. One of the most recent advances is the United States Food and Drug Administration's (FDA) approval of a rapid test. As a result of the availability of rapid diagnostic testing for HIV, practices regarding testing and counseling need to be updated, particularly in time-sensitive situations in which rapid results are crucial. PMID- 14556598 TI - Recommendations for reducing the risk of occupational HIV transmission. AB - HIV infection is an occupational risk for health care workers. To protect these health care workers, the Public Health Service has recommendations for postexposure chemoprophylaxis. PMID- 14556599 TI - HIV counseling and testing in pregnancy. PMID- 14556600 TI - Updated recommendations for reducing vertical HIV transmission. PMID- 14556601 TI - Immunizations in HIV-infected patients. PMID- 14556602 TI - Contraindicated antiretroviral drug combinations. AB - The proper implementation of combination antiretroviral treatment regimens is fundamental to successful therapeutic outcomes for patients with HIV/AIDS. Unfortunately, some patients are still being prescribed contraindicated antiretroviral regimens that include: 1. stavudine plus zidovudine; 2. Invirase plus two nucleoside analog reverse transcriptase inhibitors (NRTIS); 3. zalcitabine plus didanosine; 4. zalcitabine plus stavudine; and, 5. zalcitabine plus lamivudine. Inappropriate regimens such as these either have limited effectiveness or potential severe toxicity. PMID- 14556603 TI - HIV resistance testing. A clinical tool. AB - Drug resistance threatens to erase the recent gains made in treating HIV infection. HIV resistance testing offers clinicians the ability to screen patients for resistant strains and adjust treatment accordingly. Resistance testing is recommended for patients on antiretroviral agents with virologic failure or with suboptimal suppression of viral load after initiation of antiretroviral therapy. Because of the potential transmission of resistant strains, testing should be considered for patients with acute infection. Although the care of many patients with HIV disease is complex and should be managed by a physician with experience and expertise in HIV disease, all practitioners should be familiar with the types of testing available and their limitations. PMID- 14556604 TI - HIV and hepatitis C co-infection. AB - Infections with HIV and hepatitis C (HCV) are individually severe diseases that call for complex medical managements. HIV-HCV co-infection complicates the medical management of both diseases. This paper provides an overview of HIV-HCV co-infection with a brief review of the natural history of hepatitis C as it applies to the co-infected patient and recommendations for the diagnosis and medical management of the co-infected patient. PMID- 14556605 TI - Syringe exchange. PMID- 14556606 TI - The numbers behind the news. Helping the public understand medical research results. PMID- 14556607 TI - What do you think? PMID- 14556608 TI - Assuring the benefits of modern medicine for all Americans. PMID- 14556609 TI - Paying for quality. PMID- 14556610 TI - Blue Cross and Blue Shield of North Carolina encourages kids to "be active". PMID- 14556611 TI - A national technology assessment institute. PMID- 14556612 TI - Common ground. Health plans and physicians share information to improve processes, patient care. PMID- 14556613 TI - The Alzheimer's epidemic: preparing for the influx of dementia patients. PMID- 14556614 TI - Immunization rates climb when plans, providers, and communities collaborate. 2. PMID- 14556615 TI - Finding the solution. AAHP program aligns health plans with solutions providers. PMID- 14556616 TI - The next-generation health plan: customer responsiveness. Last of a series on health plans' evolution. PMID- 14556617 TI - The audit advantage: get paid what you deserve--on time! PMID- 14556618 TI - Provider-based clinics: opportunities and challenges. PMID- 14556619 TI - Receptor desensitization by neurotransmitters in membranes: are neurotransmitters the endogenous anesthetics? AB - A mechanism of anesthesia is proposed that addresses one of the most troubling peculiarities of general anesthesia: the remarkably small variability of sensitivity within the human population and across a broad range of animal phyla. It is hypothesized that in addition to the rapid, saturable binding of a neurotransmitter to its receptor that results in activation, the neurotransmitter also acts indirectly on the receptor by diffusing into the postsynaptic membrane and changing its physical properties, causing a shift in receptor conformational equilibrium (desensitization). Unlike binding, this slower indirect mechanism is nonspecific: each neurotransmitter will, in principle, affect all receptors in the membrane. For proteins modeled as having only resting and active conformational states, time-dependent ion currents are predicted that exhibit many characteristics of desensitization for both inhibitory and excitatory channels. If receptors have been engineered to regulate the time course of ion currents by this mechanism, then (a) mutations that significantly alter receptor sensitivity to this effect would be lethal and (b) by design, excitatory receptors would be inhibited, but inhibitory receptors activated, so that their effects are not counterproductive. The wide range of exogenous molecules that affect the physical properties of membranes as do neurotransmitters, but that do not bind to receptors, would thus inhibit excitatory channels and activate inhibitory channels, i.e., they would act as anesthesics. The endogenous anesthetics would thus be the neurotransmitters, the survival advantage conferred by their proper membrane-mediated desensitization of receptors explaining the selection pressure for anesthesic sensitivity. PMID- 14556620 TI - Attenuated total reflection Fourier transform infrared spectroscopy: a method of choice for studying membrane proteins and lipids. PMID- 14556621 TI - Presence of ferric hydroxide clusters in mutants of Haemophilus influenzae ferric ion-binding protein A. AB - The periplasmic iron binding protein plays an essential role in the iron uptake pathway of Gram-negative pathogenic bacteria from the Pasteurellaceae and Neisseriaceae families and is critical for survival of these pathogens within the host. In this study, we report the crystal structures of two mutant forms of ferric ion-binding protein A (FbpA) from Haemophilus influenzae with bound multinuclear oxo-metal clusters. Crystals of site-directed mutants in the metal or anion binding ligands contain protein in the open conformation, and two mutant FbpAs, H9A and N175L, contain different cluster arrangements in the iron-binding pocket. The iron clusters are anchored by binding to the two tyrosine ligands (Tyr195 and Tyr196) positioned at the vertex of the iron-binding pocket but are not coordinated by the other metal binding ligands. Our results suggest that the metal clusters may have formed in situ, suggesting that the mutant FbpAs may serve as a simple model for protein-mediated mineralization. PMID- 14556622 TI - PA1b, an insecticidal protein extracted from pea seeds (Pisum sativum): 1H-2-D NMR study and molecular modeling. AB - PA1b (pea albumin 1, subunit b) is a 37-amino acid cysteine-rich plant defense protein isolated from pea seeds (Pisum sativum). It induces short-term mortality in several pests, among which the cereal weevils Sitophilus sp. (Sitophilus oryzae, Sitophilus granarius, and Sitophilus zeamais) that are a major nuisance for stored cereals, all over the world. As such, PA1b is the first genuine protein phytotoxin specifically toxic to insects, which makes it a promising tool for seed weevil damage control. We have determined the 3-D solution structure of PA1b, using 2-D homonuclear proton NMR methods and molecular modeling. The primary sequence of the protein does not share similarities with other known toxins. It includes six cysteines forming three disulfide bridges. However, because of PA1b resistance to protease cleavage, conventional methods failed to establish the connectivity pattern. Our first attempts to assign the disulfide network from NOE data alone remained unsuccessful due to the tight packing of the cysteine residues within the core of the molecule. Yet, the use of ambiguous disulfide restraints within ARIA allowed us to establish that PA1b belongs to the inhibitor cystine-knot family. It exhibits the structural features that are characteristic of the knottin fold, namely, a triple-stranded antiparallel beta sheet with a long flexible loop connecting the first to the second strand and a series of turns. A comparison of the structural properties of PA1b with that of structurally related proteins adopting a knottin fold and exhibiting a diverse range of biological activities shows that the electrostatic and lipophilic potentials at the surface of PA1b are very close to those found for the spider toxin ACTX-Hi:OB4219, thereby suggesting activity on ion channels. PMID- 14556623 TI - Antitumor agents 4. Characterization of free radicals produced during reduction of the antitumor drug 5H-pyridophenoxazin-5-one: an EPR study. AB - 5H-Pyridophenoxazin-5-one (PPH), a new anticancer iminoquinone, is able to inhibit a large number of lymphoblastoid and solid tumor-derived cells at submicromolar concentrations. Molecular modeling calculations indicated that this compound might intercalate into the DNA double strand. This was also supported by nuclear magnetic resonance studies. Since free radicals arising from anticancer quinonic drugs have been proposed to be key species responsible for DNA cleavage, we have aimed to intercept and identify free radicals from PPH generated under bioreductive conditions. The first and second monoelectronic reduction potentials of PPH were measured by means of cyclic voltammetry: the reduction potential of PPH is compatible with its reduction by compounds such as NADH, and suggested that reduction of PPH may play a role in its cytotoxicity. The radical anion PPH(*)(-) was detected by means of electron paramagnetic resonance spectroscopy, and its identification was supported by DFT calculations. EPR experiments in the presence of spin traps 5,5-dimethylpyrroline N-oxide and 5-(diethoxyphosphoryl)-5 methylpyrroline N-oxide suggested the occurrence of an electron transfer between the radical anion of the drug and oxygen resulting in the formation of the superoxide anion (O(2)(*)(-)). The enthalpy of the reaction of PPH(*)(-) with O(2) was determined both in the gas phase and in solution at the B3LYP/6-31+G level using the isodensity PCM method, and the overall process in dimethyl sulfoxide was predicted to be slightly exothermic. We propose that the monoelectronic reduction of PPH in the proximity of DNA may eventually lead to radicals that could cause considerable damage to DNA, thus accounting for the high cytotoxic activity of the drug. Indeed, a comet assay (alkaline single-cell electrophoresis) showed that PPH causes free radical-induced DNA damage. PMID- 14556624 TI - NMR structures reveal how oxidation inactivates thrombomodulin. AB - Oxidation of Met 388, one of the three linker residues connecting the fourth and fifth EGF-like domains of thrombomodulin (TM), is deleterious for TM activity. An NMR structure of the smallest active fragment of TM (TMEGF45) and a crystal structure of a larger fragment (TMEGF456) bound to thrombin both show that Met 388 is packed into the fifth domain. Using multidimensional NMR, we have solved the structure of TMEGF45 in which Met 388 is oxidized (TMEGF45ox) and the structure of TMEGF45 in which Met 388 is mutated to Leu (TMEGF45ML). Comparison of the structures shows that the fifth domain has a somewhat different structure depending on the residue at position 388, and several of the thrombin-binding residues are packed into the fifth domain in the oxidized protein while they are exposed and free to interact with thrombin in the native structure and the Met Leu mutant. This observation is consistent with kinetic measurements showing that the K(m) for TMEGF45ox binding to thrombin is 3.3-fold higher than for the native protein. Most importantly, the connection between the two domains, as indicated by interdomain NOEs, appears to be essential for activity. In the TMEGF45ox structure which has a reduced k(cat) for protein C activation by the thrombin TMEGF45ox complex, interaction between the two domains is lost. Conversely, a tighter connection is observed between the two domains in TMEGF45ML, which has a higher k(cat) for protein C activation by the thrombin-TMEGF45ML complex. PMID- 14556626 TI - Glucosylation of Ras by Clostridium sordellii lethal toxin: consequences for effector loop conformations observed by NMR spectroscopy. AB - The lethal toxin (LT) from Clostridium sordellii, which belongs to the family of large clostridial cytotoxins, acts as a monoglucosyltransferase for the Rho subfamily GTPase Rac and also modifies Ras. In the present study we investigated structural changes of H-Ras in its di- and triphosphate form that occur upon glucosylation of the effector domain amino acid threonine-35 by LT. (31)P NMR experiments recorded during the enzymatic glucosylation process, using UDP glucose as a cosubstrate, show that the modification of the threonine side chain influences the chemical shifts of the phosphate groups of the bound nucleotides. In the diphosphate-bound form (Ras.GDP) glucosylation of Thr35 induces only small changes in the chemical environment of the active center. In the triphosphate form with the GTP analogue GppNHp bound (Ras.GppNHp) Ras shows at least two different conformations in the active center that exchange on a medium-range time scale (10 to 0.1 ms). Glucosylation selectively stabilizes one distinct conformation of the effector loop (state 1) with tyrosine-32 probably apart from the nucleotide and threonine-35 not involved in magnesium ion coordination. This conformation is known to have a low affinity to effector proteins such as Raf-1, AF-6, or Byr2 and thus prevents the transduction of the activation signal in the Ras-mediated pathway. NMR correlation spectra of Ras(T35glc).GDP and denaturation experiments with urea indicate that the glucose is bound in the alpha-anomeric form to the hydroxyl group of the threonine-35 side chain. Inhibition of the glucosylation reaction by 1,5-gluconolactone suggests a stereospecific reaction mechanism with a glucosyl oxonium ion transition state for the enzymatic activity of LT. PMID- 14556625 TI - Crystallographic studies on the complex behavior of nicotine binding to P450cam (CYP101). AB - Crystallographic and spectroscopic studies have been undertaken to characterize the binding behavior of the non-native substrate nicotine in the active site of the monooxygenase hemoprotein cytochrome P450cam. Despite the existence of a theoretical model that is consistent with the observed distribution of monooxygenation products, the crystal structure of the complex indicates that the primary binding mode of nicotine is unproductive. The structure is confirmed by spectral data that indicate direct coordination of substrate pyridine nitrogen with the heme iron. This would be the proper structure for evaluating binding affinity and inhibition. Reduction of the heme from Fe(III) to Fe(II) and introduction of carbon monoxide into crystals of the nicotine-P450cam complex, to simulate molecular oxygen binding, produces reorientation of the nicotine. This orientation is the appropriate one for predicting regioselectivity and the kinetic features of substrate oxidation. While it is not clear that such complicated behavior will be exhibited for other enzyme-substrate interactions, it is clear that a single crystal structure for a given substrate-enzyme interaction may not provide a good description of the binding mode responsible for product formation. PMID- 14556627 TI - Enthalpy/entropy compensation: influence of DNA flanking sequence on the binding of 7-amino actinomycin D to its primary binding site in short DNA duplexes. AB - The effect of the context of the flanking sequence on ligand binding to DNA oligonucleotides that contain consensus binding sites was investigated for the binding of the intercalator 7-amino actinomycin D. Seven self-complementary DNA oligomers each containing a centrally located primary binding site, 5'-A-G-C-T 3', flanked on either side by the sequences (AT)(n) or (AA)(n) (with n = 2, 3, 4) and AA(AT)(2), were studied. For different flanking sequences, (AA)(n)-series or (AT)(n)-series, differential fluorescence enhancements of the ligand due to binding were observed. Thermodynamic studies indicated that the flanking sequences not only affected DNA stability and secondary structure but also modulated ligand binding to the primary binding site. The magnitude of the ligand binding affinity to the primary site was inversely related to the sequence dependent stability. The enthalpy of ligand binding was directly measured by isothermal titration calorimetry, and this made it possible to parse the binding free energy into its energetic and entropic terms. Our results reveal a pronounced enthalpy-entropy compensation for 7-amino actinomycin D binding to this family of oligonucleotides and suggest that the DNA sequences flanking the primary binding site can strongly influence ligand recognition of specific sites on target DNA molecules. PMID- 14556628 TI - Electron transfer complexes of cytochrome c peroxidase from Paracoccus denitrificans containing more than one cytochrome. AB - According to the model proposed in previous papers [Pettigrew, G. W., Prazeres, S., Costa, C., Palma, N., Krippahl, L., and Moura, J. J. (1999) The structure of an electron-transfer complex containing a cytochrome c and a peroxidase, J. Biol. Chem. 274, 11383-11389; Pettigrew, G. W., Goodhew, C. F., Cooper, A., Nutley, M., Jumel, K., and Harding, S. E. (2003) Electron transfer complexes of cytochrome c peroxidase from Paracoccus denitrificans, Biochemistry 42, 2046-2055], cytochrome c peroxidase of Paracoccus denitrificans can accommodate horse cytochrome c and Paracoccus cytochrome c(550) at different sites on its molecular surface. Here we use (1)H NMR spectroscopy, analytical ultracentrifugation, molecular docking simulation, and microcalorimetry to investigate whether these small cytochromes can be accommodated simultaneously in the formation of a ternary complex. The pattern of perturbation of heme methyl and methionine methyl resonances in binary and ternary solutions shows that a ternary complex can be formed, and this is confirmed by the increase in the sedimentation coefficient upon addition of horse cytochrome c to a solution in which cytochrome c(550) fully occupies its binding site on cytochrome c peroxidase. Docking experiments in which favored binary solutions of cytochrome c(550) bound to cytochrome c peroxidase act as targets for horse cytochrome c and the reciprocal experiments in which favored binary solutions of horse cytochrome c bound to cytochrome c peroxidase act as targets for cytochrome c(550) show that the enzyme can accommodate both cytochromes at the same time on adjacent sites. Microcalorimetric titrations are difficult to interpret but are consistent with a weakened binding of horse cytochrome c to a binary complex of cytochrome c peroxidase and cytochrome c(550) and binding of cytochrome c(550) to the cytochrome c peroxidase that is affected little by the presence of horse cytochrome c in the other site. The presence of a substantial capture surface for small cytochromes on the cytochrome c peroxidase has implications for rate enhancement mechanisms which ensure that the two electrons required for re-reduction of the enzyme after reaction with hydrogen peroxide are delivered efficiently. PMID- 14556629 TI - The redox domain of the Yap1p transcription factor contains two disulfide bonds. AB - The subcellular localization of the Saccharomyces cerevisiae transcription factor Yap1p is regulated by oxidation and reduction. We purified Yap1p from yeast and characterized its properties in vitro. Electrophoretic mobility shift assays showed that the purified protein can specifically bind the TRX2 target promoter. Yap1p was purified under reducing conditions, but removal of reducing agents resulted in the formation of an oxidized Yap1p species with properties similar to in vivo oxidized Yap1p. MALDI-TOF mass spectrometry analysis revealed that the oxidized form of Yap1p contains two disulfide bonds between C303-C598 and C310 C629. A stable domain of approximately 15 kDa was detected upon limited proteolysis of oxidized but not reduced Yap1p. This Yap1p protease resistant domain was purified, and MALDI-TOF mass spectrometry analysis showed that it was comprised of two separate cysteine-containing peptides of Yap1p. These peptides are separated by 250 amino acids and are joined by the C303-C598 and C310-C629 disulfide bonds. Taken together, these data suggest that the domain that controls Yap1p subcellular localization is modular and contains a redox center comprised of four cysteine residues. PMID- 14556630 TI - Structural reorganization of proteins revealed by radiolysis and mass spectrometry: G-actin solution structure is divalent cation dependent. AB - The solution structures of isolated monomeric actins in their Mg(2+)-ATP and Ca(2+)-ATP bound forms and in complexes with gelsolin segment-1 have been probed using hydroxyl radicals (*OH) generated by synchrotron X-ray radiolysis. Proteolysis and mass spectrometry analysis of 28 peptides containing 58 distinct reactive probe sites within actin were used to monitor conformational variations linked to divalent cation and gelsolin segment-1 binding. The solvent accessibilities of the probe sites, as measured by footprinting in solution for the Ca(2+)-G-actin and Mg(2+)-G-actin complexes with gelsolin segment-1, were consistent with available crystallographic data. This included a specific protection at the contact interface between the partners, as revealed by reduced reactivity of peptide 337-359 in the complex. Aside from the specific protection indicated previously, the oxidation rates for the reactive residues of the isolated Ca(2+)-G-actin were similar to those of the actin gelsolin segment-1 complexes; however, the reactivity of numerous residues in the isolated Mg(2+)-G actin form was significantly reduced. Specifically, Mg(2+)-G-actin has a set of protected sites relative to Ca(2+)-G-actin that suggest a structural reorganization in subdomains 4 and 2 and a C-terminus more closely packed onto subdomain 1. These conformational variations for isolated Mg(2+)-G-actin provide a structural basis for its greater tendency to polymerize into filaments as compared to Ca(2+)-G-actin. PMID- 14556631 TI - Oxidative inactivation of purified plasma membrane Ca2+-ATPase by hydrogen peroxide and protection by calmodulin. AB - Calmodulin (CaM)-regulated plasma membrane Ca(2+)-ATPase (PMCA) is critical for the regulation of free intracellular Ca(2+) levels. PMCA activity and levels in neuronal membranes are decreased with aging, possibly due to oxidation-induced inactivation. In the present studies, inhibition of PMCA by H(2)O(2) was characterized in enzyme purified from human erythrocyte membranes. Basal and CaM stimulated PMCA activities were inhibited by exposure to H(2)O(2) (25-100 microM). However, neither the concentration-dependent enhancement of PMCA activity by CaM nor the binding of CaM to H(2)O(2)-exposed PMCA was disrupted by treatment with H(2)O(2). Rates of inactivation by H(2)O(2) of basal and CaM stimulated PMCA were nearly identical. The addition of CaM after exposure to H(2)O(2) did not protect enzyme activity, although the binding of CaM to PMCA before exposure to H(2)O(2) protected the enzyme completely, indicating a CaM induced conformational state resistant to oxidation. H(2)O(2) quenched Trp fluorescence in PMCA, an index of conformational changes, with a rate similar to that observed for enzyme inactivation. H(2)O(2) enhanced the solvent accessibility of Trp residues in PMCA, whereas accessibility of the only Trp residue in the CaM-binding domain peptide was unaltered. Exposure of PMCA to H(2)O(2) led to aggregate formation partially reversible by dithiothreitol (DTT) but not to recovery of activity. Amino acid analysis indicated Cys modification following H(2)O(2) exposure but no Cys oxyacids. Because DTT did not reverse inactivation by H(2)O(2), it appears that the disulfide bond formation led to conformational changes that were not fully reversed when the bonds were reduced. Preincubation of PMCA with CaM protected the enzyme from undergoing this conformational change. PMID- 14556632 TI - Modulation of kinesin half-site ADP release and kinetic processivity by a spacer between the head groups. AB - A series of modifications of the junction of the neck linker and neck coil of dimeric Drosophila kinesin were constructed to determine the influence of head orientation and spacing on the ATPase kinetics. Ala(345) is the first residue in the coiled-coil of the neck, and its replacement with glycine or proline produces no significant change in the k(cat) or K(0.5(MT)) values for activation of their ATPase by microtubules (MTs) or in their k(bi(ratio)) value for the average number of ATP molecules hydrolyzed during a processive encounter with a MT. Addition or deletion of a single amino acid at the junction produces only modest changes with less than a 2-fold reduction in kinetic processivity. Insertion of a spacer of 6 or 12 additional amino acids at the neck linker junction increases the K(0.5(MT)) value by 3-4-fold with a corresponding decrease in kinetic processivity. The sliding velocities of all the mutant constructs under multimotor conditions are within 30% of the wild-type value. All the constructs with single residue changes exhibit half-site ADP release on binding to MTs. The constructs with long insertion, however, rapidly release both ADP molecules per dimer on binding to a MT, indicating that the steric constraints that prevent release of ADP from the tethered head of wild-type kinesin have been relieved by the long insertions. The constructs with long inserts have decreased kinetic processivity and dissociate from the MT during ATP hydrolysis 3-fold faster than wild-type. PMID- 14556634 TI - Interaction of cholesterol with a docosahexaenoic acid-containing phosphatidylethanolamine: trigger for microdomain/raft formation? AB - Docosahexaenoic acid (DHA, 22:6) containing phospholipids have been postulated to be involved in promoting lateral segregation within membranes into cholesterol- (CHOL-) rich and CHOL-poor lipid microdomains. Here we investigated the specific molecular interactions of phospholipid bilayers composed of 1-[(2)H(31)]palmitoyl 2-docosahexaenoyl-sn-glycero-3-phosphoethanolamine (16:0-22:6PE-d(31)) or 1 [(2)H(31)]palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (16:0-18:1PE-d(31)) with equimolar CHOL using solid-state (2)H NMR spectroscopy and low- and wide angle X-ray diffraction (XRD). Moment analysis of (2)H NMR spectra obtained as a function of temperature reveals that the main chain melting transition and the lamellar-to-inverted hexagonal (H(II)) phase transition of 16:0-22:6PE-d(31) remain in the presence of equimolar CHOL, whereas addition of equimolar CHOL essentially obliterates the gel-to-liquid crystalline transition of 16:0-18:1PE d(31). (2)H NMR order parameter measurements show that the addition of equimolar CHOL in the lamellar liquid crystalline phase causes a smaller increase in order for the perdeuterated sn-1 chain by 22% for 16:0-22:6PE-d(31) as opposed to 33% for 16:0-18:1PE-d(31). XRD experiments determined markedly lower solubility of 32 +/- 3 mol % for CHOL in 16:0-22:6PE bilayers in contrast to the value of approximately 51 mol % for 16:0-18:1PE. Our findings provide further evidence that cholesterol has a low affinity for DHA-containing phospholipids and that this reduced affinity may serve as a mechanism for triggering the formation of lipid microdomains such as rafts. PMID- 14556633 TI - Direct analysis of tubulin expression in cancer cell lines by electrospray ionization mass spectrometry. AB - Differential expression of tubulin isotypes, mutations, and/or post-translational modifications in sensitive and Taxol-resistant cell lines suggests the existence of tubulin-based mechanisms of resistance. Since tubulin isotypes are defined by their C-terminal sequence, we previously described a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry-based analysis of tubulin diversity in human cell lines by analysis of their CNBr-released C-terminal peptides [Rao, S., Aberg, F., Nieves, E., Horwitz, S. B., and Orr, G. A. (2001) Biochemistry 40, 2096-103]. We now describe the liquid chromatography/electrospray ionization mass spectrometry analysis of native tubulins in Taxol-stabilized microtubules from parental and Taxol/epothilone resistant human cancer cell lines. This method allows the direct determination of tubulin isotype composition, including post-translational modifications and mutations occurring throughout the entire protein. Four major isotypes, betaI-, betaIVb-, Kalpha1-, and alpha6-tubulin, were detected in two human carcinoma cell lines, A549 and HeLa. betaIII-Tubulin represented a minor species, as did alpha4 tubulin which was detected for the first time in both cell lines. The three alpha tubulins were almost totally tyrosinated, and post-translational modifications were limited to low levels of monoglutamylation of Kalpha1-, betaI-, and betaIII tubulin. betaII- and betaIVa-tubulins were not detected in either parental or drug-resistant cell lines, in contrast to previous RNA-based studies. Since mutations can occur in a single tubulin allele, the question as to whether the wild-type and mutant transcripts are both translated, and to what levels, is important. Heterozygous expression of Kalpha1- or betaI-tubulin mutants that introduced mass changes as small as 26 Da was readily detected in native tubulins isolated from Taxol- and epothilone-resistant cell lines. PMID- 14556635 TI - Topological and functional study of subunit h of the F1Fo ATP synthase complex in yeast Saccharomyces cerevisiae. AB - Subunit h, a 92-residue-long, hydrophilic, acidic protein, is a component of the yeast mitochondrial F1Fo ATP synthase. This subunit, homologous to the mammalian factor F6, is essential for the correct assembly and/or functioning of this enzyme since yeast cells lacking it are not able to grow on nonfermentable carbon sources. Chemical cross-links between subunit h and subunit 4 have previously been shown, suggesting that subunit h is a component of the peripheral stalk of the F1Fo ATP synthase. The construction of cysteine-containing subunit h mutants and the use of bismaleimide reagents provided insights into its environment. Cross-links were obtained between subunit h and subunits alpha, f, d, and 4. These results and secondary structure predictions allowed us to build a structural model and to propose that this subunit occupies a central place in the peripheral stalk between the F1 sector and the membrane. In addition, subunit h was found to have a stoichiometry of one in the F1Fo ATP synthase complex and to be in close proximity to another subunit h belonging to another F1Fo ATP synthase in the inner mitochondrial membrane. Finally, functional characterization of mitochondria from mutants expressing different C-terminal shortened subunit h suggested that its C-terminal part is not essential for the assembly of a functional F1Fo ATP synthase. PMID- 14556636 TI - Vibrational Stark effects calibrate the sensitivity of vibrational probes for electric fields in proteins. AB - Infrared spectroscopy is widely used to probe local environments and dynamics in proteins. The introduction of a unique vibration at a specific site of a protein or more complex assembly offers many advantages over observing the spectra of an unmodified protein. We have previously shown that infrared frequency shifts in proteins can arise from differences in the local electric field at the probe vibration. Thus, vibrational frequencies can be used to map electric fields in proteins at many sites or to measure the change in electric field due to a perturbation. The Stark tuning rate gives the sensitivity of a vibrational frequency to an electric field, and for it to be useful, the Stark tuning rate should be as large as possible. Vibrational Stark effect spectroscopy provides a direct measurement of the Stark tuning rate and allows a quantitative interpretation of frequency shifts. We present vibrational Stark spectra of several bond types, extending our work on nitriles and carbonyls and characterizing four additional bond types (carbon-fluorine, carbon-deuterium, azide, and nitro bonds) that are potential probes for electric fields in proteins. The measured Stark tuning rates, peak positions, and extinction coefficients provide the primary information needed to design amino acid analogues or labels to act as probes of local environments in proteins. PMID- 14556637 TI - A gene cluster responsible for alkylaldoxime metabolism coexisting with nitrile hydratase and amidase in Rhodococcus globerulus A-4. AB - An enzyme "alkylaldoxime dehydratase (OxdRG)" was purified and characterized from Rhodococcus globerulus A-4, in which nitrile hydratase (NHase) and amidase coexisted with the enzyme. The enzyme contains heme b as a prosthetic group, requires reducing reagents for the reaction, and is most active at a neutral pH and at around 30 degrees C, similar to the phenylacetaldoxime dehydratase from Bacillus sp. OxB-1 (OxdB). However, some differences were seen in subunit structure, substrate specificity, and effects of activators and inhibitors. The corresponding gene, oxd, encoding a 1059-base pair ORF consisting of 353 codons, was cloned, sequenced, and overexpressed in Escherichia coli. The predicted polypeptide showed 30.3% identity to OxdB. The gene is mapped just upstream of the gene cluster encoding the enzymes involved in the metabolism of aliphatic nitriles, i.e., NHase and amidase, and their regulatory and activator proteins. We report here the existence of an aldoxime dehydratase genetically linked with NHase and amidase, and responsible for the metabolism of alkylaldoxime in R. globerulus. PMID- 14556638 TI - Characterization of Mycobacterium tuberculosis mycothiol S-conjugate amidase. AB - Mycothiol is comprised of N-acetylcysteine (AcCys) amide linked to 1D-myo inosityl 2-amino-2-deoxy-alpha-D-glucopyranoside (GlcN-Ins) and is the predominant thiol found in most actinomycetes. Mycothiol S-conjugate amidase (Mca) cleaves the amide bond of mycothiol S-conjugates of a variety of alkylating agents and xenobiotics, producing GlcN-Ins and a mercapturic acid that can be excreted from the cell. Mca of Mycobacterium tuberculosis (Rv1082) was cloned and expressed as a soluble protein in Escherichia coli. The protein contained 1.4 +/- 0.1 equiv of zinc after purification, indicating that Mca is a metalloprotein with zinc as the native metal. Kinetic studies of Mca activity with 14 substrates demonstrated that Mca is highly specific for the mycothiol moiety of mycothiol S conjugates and relatively nonspecific for the structure of the sulfur-linked conjugate. The deacetylase activity of Mca with GlcNAc-Ins is small but significant and failed to saturate at up to 2 mM GlcNAc-Ins, indicating that Mca may contribute modestly to the production of GlcN-Ins when GlcNAc-Ins levels are high. The versatility of Mca can be seen in its ability to react with a broad range of mycothiol S-conjugates, including two different classes of antibiotics. The mycothiol S-conjugate of rifamycin S was produced under physiologically relevant conditions and was shown to be a substrate for Mca in both oxidized and reduced forms. Significant activity was also seen with the mycothiol S-conjugate of the antibiotic cerulenin as a substrate for Mca. PMID- 14556639 TI - Spectroscopic studies of DNA and ATP binding to human polynucleotide kinase: evidence for a ternary complex. AB - Human polynucleotide kinase (hPNK), which possesses both 5'-DNA kinase and 3'-DNA phosphatase activities, is a DNA repair enzyme required for processing and rejoining of single- and double-strand-break termini. Full-length hPNK was subjected to sedimentation and spectroscopic analyses in association with its ligands, a 20-mer oligonucleotide, ATP, and AMP-PNP (a nonhydrolyzable analogue of ATP). Sedimentation equilibrium measurements indicated that hPNK was a monomer in the presence and absence of the ligands. Circular dichroism measurements revealed that the ligands induced different conformational changes in hPNK, although AMP-PNP induced the same conformational changes as ATP. CD also indicated that the oligonucleotide could bind to the protein-AMP-PNP complex. Protein-ligand binding affinities and stoichiometries were determined by measuring changes in protein intrinsic fluorescence. Titrating hPNK with the oligonucleotide indicated tight binding with a K(d) value of 1.3 microM and with 1:1 stoichiometry. A 5'-phosphorylated oligonucleotide with the same sequence exhibited an almost 6-fold lower affinity (K(d) value, 7.2 microM). ATP and AMP PNP bound with high affinity (K(d) values, respectively, of 1.4 and 1.6 microM), and the observed binding stoichiometries were 1:1. Furthermore, the nonphosphorylated oligonucleotide was able to bind to hPNK in the presence of AMP PNP with a K(d) value of 2.5 microM, confirming the formation of a ternary complex. This study provides the first direct physical evidence for such a ternary complex involving a polynucleotide kinase, AMP-PNP, and an oligonucleotide, and supports a reaction mechanism in which ATP and DNA bind simultaneously to the enzyme. PMID- 14556640 TI - DNA dynamics in RecA-DNA filaments: ATP hydrolysis-related flexibility in DNA. AB - RecA-catalyzed DNA recombination is initiated by a mandatory, high-energy form of DNA in RecA-nucleoprotein filaments, where bases are highly unstacked and the backbone is highly unwound. Interestingly, only the energetics consequent to adenosine triphosphate (ATP) binding, rather than its hydrolysis, seems sufficient to mediate such a high-energy structural hallmark of a recombination filament. The structural consequence of ATP hydrolysis on the DNA part of the filament thus remains largely unknown. We report time-resolved fluorescence dynamics of bases in RecA-DNA complexes and demonstrate that DNA bases in the same exhibit novel, motional dynamics with a rotational correlation time of 7-10 ns, specifically in the presence of ATP hydrolysis. When the ongoing ATP hydrolysis of RecA-DNA filament is "poisoned" by a nonhydrolyzable form of ATP (ATPgammaS), the motional dynamics cease and reveal a global motion with a rotational correlation time of >20 ns. Such ATP hydrolysis-induced flexibility ensues in single-stranded as well as double-stranded bases of RecA-DNA filaments. These results suggest that the role of ATP hydrolysis is to induce a high level of backbone flexibility in RecA-DNA filament, a dynamic property that is likely to be important for efficient strand exchanges in ATP hydrolysis specific RecA reactions. It is the absence of these motions that may cause high rigidity in RecA-DNA filaments in ATPgammaS. Dynamic light scattering measurement comparisons of RecA-ss-DNA filaments formed in ATPgammaS vs that of ATP confirmed such an interpretation, where the former showed a complex of larger (30 nm) hydrodynamic radius than that of latter (12-15 nm). Taken together, these results reveal a more dynamic state of DNA in RecA-DNA filament that is hydrolyzing ATP, which encourage us to model the role of ATP hydrolysis in RecA-mediated DNA transactions. PMID- 14556641 TI - Chirality of peptide bond-forming condensation domains in nonribosomal peptide synthetases: the C5 domain of tyrocidine synthetase is a (D)C(L) catalyst. AB - Nonribosomal peptides (NRP) such as the antibiotic tyrocidine have D-amino acids, introduced by epimerase (E) domains embedded within modules of the enzymatic assembly lines. We predict that the peptide bond-forming condensation (C) domains immediately downstream of E domains are D-specific for the peptidyl donor and L specific for the aminoacyl acceptor ((D)C(L)). To validate this prediction and establish that the C(5) domain of tyrocidine synthetase is indeed (D)C(L), the apoT (thiolation) forms of module 4 (TycB(3) AT(4)E) and module 5 (TycC(1) C(5)AT(5)) were expressed. T(5) was posttranslationally primed with CoASH to introduce the HS-pantetheinyl group and autoaminoacylated with radiolabeled L Asn* or L-Asp*. Alternate donor substrates were introduced by priming apo AT(4)E with synthetically prepared tetrapeptidyl-CoA's differing in the chirality of Phe 4, D-Phe-L-Pro-L-Phe-L-Phe-CoA, and D-Phe-L-Pro-L-Phe-D-Phe-CoA. The tetrapeptidyl-S-T(4) and L-Asp-S-T(5) were studied for peptide bond formation and chain translocation by C(5) to yield pentapeptidyl-S-T(5), whose chirality (D-L-L D-L- vs D-L-L-L-L-) was assayed by thioester cleavage and chiral chromatography of the released pentapeptides. Only the D-Phe-4 pentapeptidyl-S-T(5) was generated, implying that only D-L-L-D-S-T(4) was utilized, proving C(5) is indeed a (D)C(L) catalyst. Furthermore, a mutant with an inactive E domain transferred tetrapeptide only when loaded with D-Phe-4 tetrapeptidyl donor, not L-Phe-4, confirming that in the wild-type assembly line C(5) only transfers D-L-L-L tetrapeptidyl-S-T(4) after in situ epimerization by the E domain. These results contrast the observation that C(5) can make both L-Phe-L-Asn and D-Phe-L-Asn when assayed with Phe as the donor substrate. Hence, utilizing an aminoacyl-S-T(4) versus the natural peptidyl-S-T(4) donor produced misleading information regarding the specificity of the condensation domain. PMID- 14556642 TI - Effects of ethanol on protein kinase C alpha activity induced by association with Rho GTPases. AB - Previous studies have shown that n-alkanols have biphasic chain length-dependent effects on protein kinase C (PKC) activity induced by association with membranes or with filamentous actin [Slater, S. J., et al. (1997) J. Biol. Chem. 272, 6167 6173; Slater, S. J., et al. (2001) Biochim. Biophys. Acta 1544, 207-216]. Recently, we showed that PKCalpha is also activated by a direct membrane lipid independent interaction with Rho GTPases. Here, the effects of ethanol and 1 hexanol on Rho GTPase-induced activity were investigated using an in vitro assay system to provide further insight into the mechanism of the effects of n-alkanols on PKC activity. Both ethanol and 1-hexanol were found to have two competing concentration-dependent effects on the Ca(2+)- and phorbol ester- or diacylglycerol-dependent activities of PKCalpha associated with either RhoA or Cdc42, consisting of a potentiation at low alcohol levels and an attenuation of activity at higher levels. Measurements of the Ca(2+), phorbol ester, and diacylglycerol concentration-response curves for Cdc42-induced activation indicated that the activating effect corresponded to a shift in the midpoints of each of the curves to lower activator concentrations, while the attenuating effect corresponded to a decrease in the level of activity induced by maximal activator levels. The presence of ethanol enhanced the interaction of PKCalpha with Cdc42 within a concentration range corresponding to the potentiating effect, whereas the level of binding was unaffected by higher ethanol levels that were found to attenuate activity. Thus, ethanol may either enhance activation of PKCalpha by Rho GTPases by enhancing the interaction between the two proteins or attenuate the level of activity of Rho GTPase-associated PKCalpha by inhibiting the ensuing activating conformational change. The results also suggest that the effects of ethanol on Rho GTPase-induced activity may switch between an activation and inhibition depending on the concentration of Ca(2+) and other activators. PMID- 14556643 TI - A model for the activation of plasma membrane calcium pump isoform 4b by calmodulin. AB - Overexpression of the plasma membrane calcium pump (PMCA) isoform 4b by means of the baculovirus system enabled us, for the first time, to study the kinetics of calmodulin binding to this pump. This was done by stopped-flow fluorescence measurements using 2-chloro-(amino-Lys(75))-[6-[4-(N,N-diethylamino)phenyl]-1,3,5 triazin-4-yl]calmodulin (TA-calmodulin). Upon mixing with PMCA, the fluorescence of TA-calmodulin changed along a biphasic curve: a rapid and small increase in fluorescence was followed by a slow and large decrease that lasted about 100 s. The experiment was done at several PMCA concentrations. Global fitting nonlinear regression analysis of these results led to a model in which PMCA is present in two forms: a closed conformation and an open conformation. Calmodulin reacts with both conformations but reacts faster and with higher affinity for the open conformation. Measurements of the ATPase activity of PMCA under similar conditions revealed that the open form has higher ATPase activity than the closed one. Contrasting with the reaction with the whole pump, TA-calmodulin reacted rapidly (in about 2 s) with a calmodulin-binding peptide made after the sequence of the calmodulin-binding domain of PMCA (C28). Results of TA-calmodulin binding to C28 are explained by a simpler model, in which only an open conformation exists. PMID- 14556644 TI - Effects of delayed preconditioning on myocardial regional contractility during repeated episodes of low-flow ischaemia in anaesthetized dogs: possible role of nitric oxide. AB - The effect of cardiac pacing on repeated low-flow ischaemia-induced changes in regional myocardial segmental contractility, and the role in these changes of nitric oxide, were investigated in anaesthetized dogs. Dogs were instrumented for cardiac pacing (pacing electrode in the right ventricle). Dogs were paced (four times for 5 min; pacing rate 220 beats.min(-1)) 24 h prior to the repeated ischaemic insults. Controls were instrumented, but not paced. After 24 h, the dogs were re-anaesthetized with pentobarbitone and subjected to four 20 min low flow ischaemia and reperfusion cycles, by constricting the left anterior descending coronary artery (LAD) to achieve an approx. 50% reduction in resting coronary blood flow. In some dogs (both control and paced), N (G)-nitro-L arginine methyl ester (L-NAME; a non-selective inhibitor of nitric oxide synthase) was infused into a side-branch of the LAD 10 min prior to the first ischaemia/reperfusion cycle. Regional contractile function was measured by ultrasonic microcrystals in the ischaemic and normal regions of the left ventricular wall supplied from the LAD and left circumflex coronary artery respectively, and expressed as percentage changes in segmental shortening (%SS). In some dogs, myocardial tissue blood flow (coloured microspheres) and lactate production (local coronary venous sampling) were measured; samples were also taken for histological analysis. In control dogs, the regional %SS was progressively reduced within the ischaemic segment during the four repeated occlusions (by 40+/-6, 59+/-6, 68+/-6, 70+/-6% during occlusions 1-4 respectively). These reductions were more pronounced, especially during the first two cycles (68+/-6, 68+/-6, 67+/-6, 67+/-6%, respectively), when the dogs had been previously subjected to cardiac pacing. In both paced and control dogs, these changes in contractile function were L-NAME-sensitive. Thus, in the presence of L-NAME, changes in regional segmental shortening in control dogs were 37+/-8, 40+/-8, 37+/-8, 42+/-11% and in the paced dogs 46+/-6, 45+/-7, 45+/-8, 45+/-7% respectively, during the four consecutive occlusions. There were no significant differences in tissue blood flow or in lactate production between the groups, and no structural changes indicative of infarction. These results show that the myocardium rapidly adapts to re-occurring acute ischaemia by reducing contractility within the ischaemic segment and, thereby, metabolic demand. Furthermore, cardiac pacing 24 h prior to these ischaemic challenges induces a similar adaptive response, a form of 'delayed preconditioning'. Since both the acute and delayed adaptation were L-NAME-sensitive, we suggest that this adaptation involves nitric oxide. PMID- 14556645 TI - Oxidative stress-related parameters in the liver of non-alcoholic fatty liver disease patients. AB - Oxidative stress is implicated in the pathogenesis of non-alcoholic fatty liver disease (NAFLD). In the present study, hepatic and plasma oxidative stress related parameters were measured and correlated with clinical and histological findings in 31 NAFLD patients showing increased body mass index. Liver protein carbonyl content was enhanced by 403% in patients with steatosis (n=15) compared with control values (n=12), whereas glutathione content, superoxide dismutase (SOD) activity and the ferric reducing ability of plasma (FRAP) were decreased by 57%, 48% and 21% (P<0.05) respectively. No changes in microsomal p-nitrophenol hydroxylation and the total content of cytochrome P450 (CYP) or CYP2E1 were observed. Patients with steatohepatitis (n=16) exhibited protein carbonyl content comparable with that of controls, whereas glutathione content, SOD and catalase activities were decreased by 27%, 64% and 48% (P<0.05). In addition, FRAP values in patients with steatohepatitis were reduced by 33% and 15% (P<0.05) when compared with controls and patients with steatosis respectively, whereas p nitrophenol hydroxylation (52%) and CYP2E1 content (142%) were significantly increased (P<0.05) compared with controls. It is concluded that oxidative stress is developed in the liver of NAFLD patients with steatosis and is exacerbated further in patients with steatohepatitis, which is associated with CYP2E1 induction. Substantial protein oxidation is followed by proteolysis of the modified proteins, which may explain the co-existence of a diminished antioxidant capacity and protein oxidation in the liver of patients with steatohepatitis. PMID- 14556646 TI - Rosiglitazone, an agonist of peroxisome-proliferator-activated receptor gamma (PPARgamma), decreases inhibitory serine phosphorylation of IRS1 in vitro and in vivo. AB - Peroxisome-proliferator-activated receptor gamma agonists such as rosiglitazone, a thiazolidinedione, improve insulin sensitivity in vivo, but the underlying mechanism(s) remains unclear. Phosphorylation of IRS1 (insulin receptor substrate protein 1) on certain serine residues, including S307 and S612 in rodent IRS1 (equivalent to S312 and S616 in human IRS1), has been shown to play a negative role in insulin signalling. In the present study, we investigated whether rosiglitazone improves insulin sensitivity by decreasing IRS1 inhibitory serine phosphorylation. In HEK-293 (human embryonic kidney 293) cells stably expressing recombinant IRS1 and in 3T3L1 adipocytes, rosiglitazone attenuated PMA-induced IRS1 S307/S612 phosphorylation and decreased insulin-stimulated Akt phosphorylation. We observed increased IRS1 S307 phosphorylation and concomitant decrease in insulin signalling as measured by insulin-stimulated IRS1 tyrosine phosphorylation, and Akt threonine phosphorylation in adipose tissues of Zucker obese rats compared with lean control rats. Treatment with rosiglitazone at 30 mg/kg body weight for 24 and 48 h increased insulin signalling and decreased IRS1 S307 phosphorylation concomitantly. Whereas the 48 h treatment reversed hyper phosphorylation (and activation) of both c-Jun N-terminal kinase and p38 mitogen activated protein kinase, the 24 h treatments only decreased hyper phosphorylation of p38 mitogen-activated protein kinase. The treatment of the Zucker obese rats with rosiglitazone also reversed the high circulating levels of non-esterified fatty acids, which have been shown to be correlated with increased IRS1 serine phosphorylation in other animal models. Taken together, these results suggest that IRS1 inhibitory serine phosphorylation is a key component of insulin resistance and its reversal contributes to the insulin sensitizing effects by rosiglitazone. PMID- 14556647 TI - Identification, characterization and subcellular localization of TcPDE1, a novel cAMP-specific phosphodiesterase from Trypanosoma cruzi. AB - Compartmentalization of cAMP phosphodiesterases plays a key role in the regulation of cAMP signalling in mammals. In the present paper, we report the characterization and subcellular localization of TcPDE1, the first cAMP-specific phosphodiesterase to be identified from Trypanosoma cruzi. TcPDE1 is part of a small gene family and encodes a 929-amino-acid protein that can complement a heat shock-sensitive yeast mutant deficient in phospho-diesterase genes. Recombinant TcPDE1 strongly associates with membranes and cannot be released with NaCl or sodium cholate, suggesting that it is an integral membrane protein. This enzyme is specific for cAMP and its activity is not affected by cGMP, Ca2+, calmodulin or fenotiazinic inhibitors. TcPDE1 is sensitive to the phosphodiesterase inhibitor dipyridamole but is resistant to 3-isobutyl-1-methylxanthine, theophylline, rolipram and zaprinast. Papaverine, erythro-9-(2-hydroxy-3-nonyl) adenine hydrochloride, and vinpocetine are poor inhibitors of this enzyme. Confocal laser scanning of T. cruzi epimastigotes showed that TcPDE1 is associated with the plasma membrane and concentrated in the flagellum of the parasite. The association of TcPDE1 with this organelle was confirmed by subcellular fractionation and cell-disruption treatments. The localization of this enzyme is a unique feature that distinguishes it from all the trypanosomatid phosphodiesterases described so far and indicates that compartmentalization of cAMP phosphodiesterases could also be important in these parasites. PMID- 14556648 TI - N-Formyl peptide receptor subtypes in human neutrophils activate L-plastin phosphorylation through different signal transduction intermediates. AB - We investigated the coupling of the fMLP (N -formyl-L-methionyl-L-leucyl-L phenylalanine; 'chemotactic peptide') receptor with phosphorylation of the actin binding protein L-plastin in neutrophils. Using two-dimensional IEF (isoelectric focusing)/PAGE and MALDI-TOF (matrix-assisted laser desorption ionization-time-of flight)-MS, L-plastin was identified as a major phosphoprotein in fMLP-stimulated neutrophils whose phosphorylation was dependent on phosphoinositide 3-kinase, PLD (phospholipase D) and PKC (protein kinase C) activity. Two fMLP receptor subtypes were identified in neutrophils, characterized by a distinct sensitivity to fMLP and antagonistic peptides. Both receptor subtypes induced the phosphorylation of L-plastin. L-plastin phosphorylation induced by low-affinity fMLP receptors involves an action of phosphoinositide 3-kinase, PLD and PKC isotypes. In contrast, none of these intermediates are utilized by high-affinity fMLP receptors in the phosphorylation of L-plastin. However, the PKC inhibitor Ro-31 8220 inhibits L-plastin phosphorylation induced by the high-affinity fMLP receptor. Thus, an as yet unknown Ro-31-8220-sensitive kinase regulates L-plastin phosphorylation in response to the high-affinity fMLP receptor. The results suggest a model in which receptor subtypes induce a similar endpoint event through different signal-transduction intermediates. This may be relevant in the context of cell migration in which one receptor subpopulation may become desensitized in a concentration gradient of chemoattractant. PMID- 14556649 TI - Calcium/calmodulin-dependent protein kinase IIdelta associates with the ryanodine receptor complex and regulates channel function in rabbit heart. AB - Cardiac ryanodine receptors (RyR2s) play a critical role in excitation contraction coupling by providing a pathway for the release of Ca(2+) from the sarcoplasmic reticulum into the cytosol. RyR2s exist as macromolecular complexes that are regulated via binding of Ca(2+) and protein phosphorylation/dephosphorylation. The present study examined the association of endogenous CaMKII (calcium/calmodulin-dependent protein kinase II) with the RyR2 complex and whether this enzyme could modulate RyR2 function in isolated rabbit ventricular myocardium. Endogenous phosphorylation of RyR2 was verified using phosphorylation site-specific antibodies. Co-immunoprecipitation studies established that RyR2 was physically associated with CaMKIIdelta. Quantitative assessment of RyR2 protein was performed by [(3)H]ryanodine binding to RyR2 immunoprecipitates. Parallel kinase assays allowed the endogenous CaMKII activity associated with these immunoprecipitates to be expressed relative to the amount of RyR2. The activity of RyR2 in isolated cardiac myocytes was measured in two ways: (i) RyR2-mediated Ca(2+) release (Ca(2+) sparks) using confocal microscopy and (ii) Ca(2+)-sensitive [(3)H]ryanodine binding. These studies were performed in the presence and absence of AIP (autocamtide-2-related inhibitory peptide), a highly specific inhibitor of CaMKII. At 1 microM AIP Ca(2+) spark duration, frequency and width were decreased significantly. Similarly, 1 microM AIP decreased [(3)H]ryanodine binding. At 5 microM AIP, a more profound inhibition of Ca(2+) sparks and a decrease in [(3)H]ryanodine binding was observed. Separate measurements showed that AIP (1-5 microM) did not affect sarcoplasmic reticulum Ca(2+)-ATPase-mediated Ca(2+) uptake. These results suggest the existence of an endogenous CaMKIIdelta that associates directly with RyR2 and specifically modulates RyR2 activity. PMID- 14556650 TI - Evidence for specific tetraspanin homodimers: inhibition of palmitoylation makes cysteine residues available for cross-linking. AB - It is a well-established fact that tetraspanin proteins, a large family of integral membrane proteins involved in cell motility, fusion and signalling, associate extensively with one another and with other transmembrane and membrane proximal proteins. In this study, we present results strongly suggesting that tetraspanin homodimers are fundamental units within larger tetraspanin complexes. Evidence for constitutive CD9 homodimers was obtained using several cell lines, utilizing the following four methods: (1) spontaneous cross-linking via intermolecular disulphide bonds, (2) use of a cysteine-reactive covalent cross linking agent, (3) use of an amino-reactive covalent cross-linking agent, and (4) covalent cross-linking via direct intermolecular disulphide bridging between unpalmitoylated membrane-proximal cysteine residues. In the last case, incubation of cells with the palmitoylation inhibitor 2-bromopalmitate exposed membrane proximal cysteine residues, thus effectively promoting 'zero-length' cross linking to stabilize homodimers. Similar to CD9, other tetraspanins (CD81 and CD151) also showed a tendency to homodimerize. Tetraspanin homodimers were assembled from newly synthesized proteins in the Golgi, as evidenced by cycloheximide and Brefeldin A inhibition studies. Importantly, tetraspanin homodimers appeared on the cell surface and participated in typical 'tetraspanin web' interactions with other proteins. Whereas homodimers were the predominant cross-linked species, we also observed some higher-order complexes (trimers, tetramers or higher) and a much lower level of cross-linking between different tetraspanins (CD81-CD9, CD9-CD151, CD81-CD151). In conclusion, our results strongly suggest that tetraspanin homodimers, formed in the Golgi and present at the cell surface, serve as building blocks for the assembly of larger, multicomponent tetraspanin protein complexes. PMID- 14556653 TI - Computer-aided tissue engineering: application to biomimetic modelling and design of tissue scaffolds. AB - Computer-aided tissue engineering (CATE) enables many novel approaches in modelling, design and fabrication of complex tissue substitutes with enhanced functionality and improved cell-matrix interactions. Central to CATE is its bio tissue informatics model that represents tissue biological, biomechanical and biochemical information that serves as a central repository to interface design, simulation and tissue fabrication. The present paper discusses the application of a CATE approach to the biomimetic design of bone tissue scaffold. A general CATE based process for biomimetic modelling, anatomic reconstruction, computer assisted-design of tissue scaffold, quantitative-computed-tomography characterization, finite element analysis and freeform extruding deposition for fabrication of scaffold is presented. PMID- 14556651 TI - Development of a novel fluorogenic proteolytic beacon for in vivo detection and imaging of tumour-associated matrix metalloproteinase-7 activity. AB - The present study describes the in vivo detection and imaging of tumour associated MMP-7 (matrix metalloproteinase-7 or matrilysin) activity using a novel polymer-based fluorogenic substrate PB-M7VIS, which serves as a selective 'proteolytic beacon' (PB) for this metalloproteinase. PB-M7VIS is built on a PAMAM (polyamido amino) dendrimer core of 14.2 kDa, covalently coupled with an Fl (fluorescein)-labelled peptide Fl(AHX)RPLALWRS(AHX)C (where AHX stands for aminohexanoic acid) and with TMR (tetramethylrhodamine). PB-M7VIS is efficiently and selectively cleaved by MMP-7 with a k (cat)/ K (m) value of 1.9x10(5) M( 1).s(-1) as measured by the rate of increase in Fl fluorescence (up to 17-fold for the cleavage of an optimized PB-M7VIS) with minimal change in the TMR fluorescence. The K (m) value for PB-M7VIS is approx. 0.5 microM, which is approx. two orders of magnitude lower when compared with that for an analogous soluble peptide, indicating efficient interaction of MMP-7 with the synthetic polymeric substrate. With MMP-2 or -3, the k (cat)/ K (m) value for PB-M7VIS is approx. 56- or 13-fold lower respectively, when compared with MMP-7. In PB-M7VIS, Fl(AHX)RPLALWRS(AHX)C is a selective optical sensor of MMP-7 activity and TMR serves to detect both the uncleaved and cleaved reagents. Each of these can be visualized as subcutaneous fluorescent phantoms in a mouse and optically discriminated based on the ratio of green/red (Fl/TMR) fluorescence. The in vivo specificity of PB-M7VIS was tested in a mouse xenograft model. Intravenous administration of PB-M7VIS gave significantly enhanced Fl fluorescence from MMP-7 positive tumours, but not from control tumours ( P <0.0001), both originally derived from SW480 human colon cancer cells. Prior systemic treatment of the tumour-bearing mice with an MMP inhibitor BB-94 ([4-( N -hydroxyamino)-2 R isobutyl-3 S -(thienylthiomethyl)-succinyl]-L-phenylalanine- N -methylamide), markedly decreased the Fl fluorescence over the MMP-7-positive tumour by approx. 60%. Thus PB-M7VIS functions as a PB for in vivo detection of MMP-7 activity that serves to light this optical beacon and is, therefore, a selective in vivo optical molecular imaging contrast reagent. PMID- 14556652 TI - Investigation of metal binding and activation of Escherichia coli glyoxalase I: kinetic, thermodynamic and mutagenesis studies. AB - GlxI (glyoxalase I) isomerizes the hemithioacetal formed between glutathione and methylglyoxal. Unlike other GlxI enzymes, Escherichia coli GlxI exhibits no activity with Zn(2+) but maximal activation with Ni(2+). To elucidate further the metal site in E. coli GlxI, several approaches were undertaken. Kinetic studies indicate that the catalytic metal ion affects the k (cat) without significantly affecting the K (m) for the substrate. Inductively coupled plasma analysis and isothermal titration calorimetry confirmed one metal ion bound to the enzyme, including Zn(2+), which produces an inactive enzyme. Isothermal titration calorimetry was utilized to determine the relative binding affinity of GlxI for various bivalent metals. Each metal ion examined bound very tightly to GlxI with an association constant ( K (a))>10(7) M(-1), with the exception of Mn(2+) ( K (a) of the order of 10(6) M(-1)). One of the ligands to the catalytic metal, His(5), was altered to glutamine, a side chain found in the Zn(2+)-active Homo sapiens GlxI. The affinity of the mutant protein for all bivalent metals was drastically decreased. However, low levels of activity were now observed for Zn(2+)-bound GlxI. Although this residue has a marked effect on metal binding and activation, it is not the sole factor determining the differential metal activation between the human and E. coli GlxI enzymes. PMID- 14556654 TI - Laryngeal Involvement of Rhabdomyosarcoma in an Adult. AB - BACKGROUND: Although initial presentation of sarcomas in the head and neck region is relatively common, involvements of the larynx by rhabdomyosarcoma either primary or metastatic have been reported rarely. CASE REPORT: A case of rhabdomyosarcoma in the right thigh, which involved laryngeal structures three years after the diagnosis, is presented. CONCLUSIONS: The laryngeal involvement by primary and metastatic rhabdosarcomas is very rare, however when it occurs it can result in life treating upper airway obstruction. After emergency intervention, radiotherapy and / or chemotherapy must be undertaken for long-term benefit and surgery may be helpful only in selected cases. PMID- 14556655 TI - Role of AML1/Runx1 in the pathogenesis of hematological malignancies. AB - AML1/Runx1, originally identified as a gene located at the breakpoint of the t(8;21) translocation, encodes one of the two subunits forming a heterodimeric transcription factor. AML1 contains a highly evolutionally conserved domain called the Runt domain, responsible for both DNA binding and heterodimerization with the partner protein, CBFbeta. AML1 is widely expressed in all hematopoietic lineages, and regulates the expression of a variety of hematopoietic genes. Numerous studies have shown that AML is a critical regulator of hematopoietic development. In addition, AML1 and CBFbeta are frequent targets for chromosomal translocation in human leukemia. Translocations lead to the generation of fusion proteins, which play a causative role for the development of leukemia, primarily by inhibiting AML1 function. Point mutations that impair AML1 function are also associated with familial and sporadic leukemias. Loss of AML1 function is thus implicated in a number of leukemias through multiple pathogenic mechanisms. However, AML1-related translocations or haploinsufficiency of AML1 are not immediately leukemogenic in animal models, suggesting that additional genetic events are required for the development of full-blown leukemia. PMID- 14556656 TI - Pre-B lymphomas in SL/Kh mice: a multifactorial disease model. AB - The pre-B lymphoma in the inbred mouse strain SL/Kh is an excellent multifactorial disease model. The endogenous murine leukemia virus provirus Emv11, genetically acquired from an AKR progenitor, is the pathogenetic agent that reintegrates to dysregulate several host genes, i.e., Stat5a, Evi3, c-Myc, N Myc, Stat5b, and others. Constitutive activation of Stat5a either by provirus integration or by transfection of the active mutant Stat5a cDNA transforms pre-B cells in bone marrow. Genetically determined expansion of early B cells and a dominant SL/Kh MHC allele predispose the animals to succumb to pre-B lymphomas. A number of other host genetic and epigenetic factors that determine the types of lymphomas, susceptibility to lymphomas, and length of the latent period are discussed. PMID- 14556657 TI - Cytokeratin 19 expression in hepatocellular carcinoma predicts early postoperative recurrence. AB - Clinicopathologic features and postoperative outcomes were investigated for patients who underwent curative surgery for biliary marker (CK7 and CK19) positive hepatocellular carcinoma (HCC). Of 157 HCCs, 93 were CK7(-)CK19(-), 49 were CK7(+)-CK19(-), 1 was CK7(-)CK19(+), and 14 were CK7(+)- CK19(+). Semiquantitative analysis of expression levels demonstrated a significant correlation between CK7 and CK19 expression. Of various clinicopathologic parameters, tumor differentiation exhibited a significant correlation with CK7 and CK19 expression. All 15 patients with CK19-positive HCC also had anti-HBc. Log-rank test revealed that CK7 expression, CK19 expression, high aspartate aminotransferase (AST) activity, low albumin concentration, portal invasion, intrahepatic metastasis, and severe fibrosis (cirrhosis) reduced the tumor-free survival rate. Multivariate analysis demonstrated that CK19 expression, intrahepatic metastasis, and severe fibrosis were independent predictors of postoperative recurrence, while CK7 expression was not. Twelve of 15 patients with CK19-positive HCC had tumor recurrence within 2 years after surgery, a significantly higher incidence of early recurrence than for CK19-negative HCC. The incidence of extrahepatic disease, especially lymph node metastasis, was significantly higher for patients with CK19-positive HCC. These findings indicate that CK19 expression is a predictor of early postoperative recurrence due to increased invasiveness. PMID- 14556658 TI - Establishment and characterization of unique cell lines derived from pyothorax associated lymphoma which develops in long-standing pyothorax and is strongly associated with Epstein-Barr virus infection. AB - Malignant lymphoma frequently develops in the pleural cavity of patients with over 20 years' history of pyothorax. The term pyothorax-associated lymphoma (PAL) has been proposed for this type of tumor. We established four novel lymphoma cell lines (OPL-3, -4, -5, and -7) from four patients with PAL. Characteristics of the four cell lines are as follows: B-cell nature with defective expression of B-cell and T-cell surface antigens, monoclonal pattern of Epstein-Barr virus (EBV) infection in lymphoma cells (thus indicating an etiological role of EBV for lymphomagenesis), complicated chromosomal abnormalities with numerous structural and numerical abnormalities, and occasional but distinct genome instability. These abnormalities in cell character might be caused by the specific circumstances of PAL lymphomagenesis, i.e., chronic inflammation. Thus, PAL cell lines could be useful in analysis of molecular mechanisms leading to malignancy in chronic inflammation. PMID- 14556659 TI - Ubiquitin-immunoreactive degradation products of cytokeratin 8/18 correlate with aggressive breast cancer. AB - Decreased amounts of cytokeratin (CK) 8/18 in the cytoplasm of breast cancer cells correlate with a poor prognosis. Although such decreases have been attributed to suppressed gene expression, accelerated protein degradation may also be responsible. In order to investigate whether selective degradation via the ubiquitin (Ub)-dependent proteasome pathway occurs in breast cancer, one- and two-dimensional (1-D and 2-D) immunoblot analysis was performed on cancerous and normal breast tissue from 50 breast cancer patients using the anti-Ub monoclonal antibodies (mAbs) KM691 and KM690. On 1-D gel electrophoresis, one broad band or two bands were detected at about 43 kDa; these were detected only in cancer tissue. Immunoreactive bands at 43 kDa were significantly associated with aggressive morphology (P = 0.011), nuclear p53 accumulation (P = 0.015) and overexpression of Her2 / neu protein (P = 0.012). On 2-D gel electrophoresis, these bands were fractionated into a group of several spots that formed a staircase pattern at 40-45 kDa. Partial amino acid sequencing analysis demonstrated that these Ub-immunoreactive spots corresponded to CK8 and CK18; however, since they did not have an amino-terminal domain, and were located at lower molecular weight positions than intact CK8 and CK18 on the 2-D gel, they were regarded as degradation products. CK18 degradation was confirmed by confocal microscopy as loss of the frame-like network that forms the luminal structure. These results indicate that CK 8/18 degradation products are detected specifically in breast cancer and may determine its aggressiveness. PMID- 14556660 TI - Estrogen sulfotransferase and sulfatase: Roles in the regulation of estrogen activity in human uterine endometrial carcinomas. AB - The regulation of estrogen activity through the formation and cleavage of sulfoconjugates of estrogens is known to be related to the progression and metastasis of estrogen-dependent breast carcinomas, but the involvement of sulfoconjugates in the steroid stimulation of endometrial functions and the progression of endometrial adenocarcinomas is not clearly understood yet. Estrogen sulfotransferase (EST) in the uterine endometria during the follicular phase was more active than during the luteal phase, but estrogen sulfate (ES) sulfatase exhibited lower activity during the follicular phase than during the luteal phase. However, ES sulfatase activities in cancerous tissues were lower than those in normal endometria and endometrial adenocarcinoma-derived cells, among which the activity was exceedingly high in Ishikawa cells, suggesting that ES sulfatase in Ishikawa cells contributes to the estrogen-dependent growth of these cells. EST activities higher than that in Ishikawa cells were found in only 3 of 24 cancerous tissues. Reverse transcriptase-polymerase chain reaction (RT PCR) analysis of the EST and ES sulfatase genes in carcinoma-derived cells demonstrated the extensive expression of both genes in Ishikawa cells. The isolated EST gene was transfected into Ishikawa cells with a mammalian expression vector to establish cell clones with enhanced EST activity, and the estrogen dependent cell growth of the resultant cell clones was found to be abolished, due to the enhanced sulfoconjugation of estrogen. Since ES sulfatase activity in cancerous tissues was significantly lower than that in Ishikawa cells, it might be not involved in the enhancement of estrogen activity associated with the pathogenesis of endometrial adenocarcinoma tissues. PMID- 14556661 TI - Development of consensus fluorogenically labeled probes of the immunoglobulin heavy-chain gene for detecting minimal residual disease in B-cell non-Hodgkin lymphomas. AB - We have examined 72 patients with B-cell non-Hodgkin lymphoma (B-NHL) in order to search for consensus sequences of the immunoglobulin heavy chain (IgH) gene, and developed consensus fluorogenically labeled probes for use in an allele-specific oligonucleotide (ASO) real-time quantitative polymerase chain reaction (RQ-PCR) assay of minimal residual disease (MRD). We detected a clonal IgH variable region (VH) sequence in 51 (70.8%) of the 72 B-NHLs, the most frequent VH gene usages being VH3 and VH4 (45/51, 88.2%). It was possible to design three consensus fluorogenic probes for the VH3 gene and one for the VH4 gene avoiding these hypermutations. Our sequencing results suggested that consensus fluorogenic probes would be best based on the 5'-side of the framework region 3 (FR3) because the frequency of somatic hypermutations was significantly lower in the regions on which the probes were based than in the remaining parts of FR3 (P < 0.05). Nineteen (54.3%) of 35 B-NHLs with the VH3 gene and 5 (50%) of 10 with the VH4 gene had sequences identical to at least one of these probes. We found that probes containing one base substitution were still applicable for a MRD study, whereas those containing two or more were not. Therefore, our four probes were applicable for 37 (82.2%) of the 45 patients with VH3 or VH4. This limited number of probes makes a large-scale study of MRD in B-NHL more feasible. PMID- 14556662 TI - Characterization of cell death induced by ethacrynic acid in a human colon cancer cell line DLD-1 and suppression by N-acetyl-L-cysteine. AB - Since ethacrynic acid (EA), an SH modifier as well as glutathione S-transferase (GST) inhibitor, has been suggested to induce apoptosis in some cell lines, its effects on a human colon cancer cell line DLD-1 were examined. EA enhanced cell proliferation at 20-40 microM, while it caused cell death at 60-100 microM. Caspase inhibitors did not block cell death and DNA ladder formation was not detected. Poly(ADP-ribose) polymerase, however, was cleaved into an 82-kDa fragment, different from an 85-kDa fragment that is specific for apoptosisis. The 82-kDa fragment was not recognized by antibody against PARP fragment cleaved by caspase 3. N-Acetyl-L-cysteine (NAC) completely inhibited EA-induced cell death, but 3(2)-t-butyl-4-hydroxyanisole or pyrrolidinedithiocarbamate ammonium salt did not. Glutathione (GSH) levels were dose-dependently increased in cells treated with EA and this increase was hardly affected by NAC addition. Mitogen-activated protein kinase (MAPK) kinase (MEK) 1, extracellular signal-regulated kinase (ERK) 1 and GST P1-1 were increased in cells treated with 25-75 microM EA, while c-Jun N-terminal kinase (JNK) 1 and p38 MAPK were markedly decreased by 100 microM EA. NAC repressed EA-induced alterations in these MAPKs and GST P1-1. p38 MAPK inhibitors, SB203580 and FR167653, dose-dependently enhanced EA-induced cell death. An MEK inhibitor, U0126, did not affect EA-induced cell death. These studies revealed that EA induced cell death concomitantly with a novel PARP fragmentation, but without DNA fragmentation. p38 MAPK was suggested to play an inhibitory role in EA-induced cell death. PMID- 14556663 TI - Up-regulation of DNA-dependent protein kinase correlates with radiation resistance in oral squamous cell carcinoma. AB - DNA-PK is a nuclear protein with serine/threonine kinase activity and forms a complex consisting of the DNA-PKcs and a heterodimer of Ku70 and Ku80 proteins. Recent laboratory experiments have demonstrated that the DNA-PK complex formation is one of the major pathways by which mammalian cells respond to DNA double strand breaks induced by ionizing radiation. In this study, we evaluated the relationship between expression levels of DNA-PKcs, Ku70 and Ku80 proteins and radiation sensitivity in oral squamous cell carcinoma (OSCC) cell lines and in OSCC patients treated with preoperative radiation therapy. The OSCC cell lines greatly differed in their response to irradiation, as assessed by a standard colony formation assay. However, the expression levels of the DNA-PK complex proteins were all similar, and there was no association between the magnitude of their expression and the tumor radiation sensitivity. Expression of DNA-PK complex proteins increased after radiation treatment, and the increased values correlated with the tumor radiation resistance. Expression of DNA-PKcs and Ku70 after irradiation was increased in the surviving cells of OSCC tissues irradiated preoperatively. These results suggest that up-regulation of DNA-PK complex protein, especially DNA-PKcs, after radiation treatment correlates to radiation resistance. DNA-PKcs might be a molecular target for a novel radiation sensitization therapy of OSCC. PMID- 14556664 TI - DNA methylation of multiple genes in gastric carcinoma: association with histological type and CpG island methylator phenotype. AB - Hypermethylation of CpG islands is associated with silencing of various tumor suppressor genes. Recent studies on colorectal and gastric cancer have identified a CpG island methylator phenotype (CIMP), which involves the targeting of multiple genes by promoter hypermethylation. For determination of association between DNA methylation pattern or histological type and CIMP status in gastric carcinoma, CpG islands in the promoters of hMLH1 and CDH1 genes, CpG islands overlapping exon 1 of MGMT and p16(INK4a) genes, and a non-CpG island in exon 1 of the RAR-beta gene were studied. The presence of the CIMP was determined by monitoring five methylated in tumor (MINT ) loci in 103 gastric carcinomas. Among the 103 gastric carcinomas, DNA hypermethylation was detected in the following frequencies: 14 (14%) for hMLH1, 26 (25%) for MGMT, 26 (25%) for p16(INK4a), 54 (52%) for CDH1, and 53 (52%) for RAR-beta. Forty-two (41%) of 103 gastric carcinomas were positive for the CIMP. CIMP and hypermethylation of p16(INK4a) gene were found more frequently in intestinal and diffuse-adherent types than in diffuse-scattered type (P = 0.013 and 0.017, respectively). In contrast, hypermethylation of the CDH1 and RAR-beta genes was more common in the diffuse scattered type than in the other types (P = 0.008 and 0.007, respectively). In intestinal- and diffuse-adherent-type gastric carcinomas, we found significant associations between the presence of the CIMP and hypermethylation of several genes: hMLH1 (P = 0.006), p16(INK4a) (P = 0.018), CDH1 (P = 0.024), and RAR-beta (P = 0.044). Our overall results suggest that in some intestinal- and diffuse adherent-type gastric carcinomas, DNA hypermethylation affects non-specific gene promoters concordantly, at least in part, whereas in diffuse-scattered-type gastric carcinoma, DNA hypermethylation affects specific genes such as CDH1 and RAR-beta. PMID- 14556665 TI - Multiple fuzzy neural network system for outcome prediction and classification of 220 lymphoma patients on the basis of molecular profiling. AB - A fuzzy neural network (FNN) using gene expression profile data can select combinations of genes from thousands of genes, and is applicable to predict outcome for cancer patients after chemotherapy. However, wide clinical heterogeneity reduces the accuracy of prediction. To overcome this problem, we have proposed an FNN system based on majoritarian decision using multiple noninferior models. We used transcriptional profiling data, which were obtained from "Lymphochip" DNA microarrays (http://llmpp.nih.gov/DLBCL), reported by Rosenwald (N Engl J Med 2002; 346: 1937-47). When the data were analyzed by our FNN system, accuracy (73.4%) of outcome prediction using only 1 FNN model with 4 genes was higher than that (68.5%) of the Cox model using 17 genes. Higher accuracy (91%) was obtained when an FNN system with 9 noninferior models, consisting of 35 independent genes, was used. The genes selected by the system included genes that are informative in the prognosis of Diffuse large B-cell lymphoma (DLBCL), such as genes showing an expression pattern similar to that of CD10 and BCL-6 or similar to that of IRF-4 and BCL-4. We classified 220 DLBCL patients into 5 groups using the prediction results of 9 FNN models. These groups may correspond to DLBCL subtypes. In group A containing half of the 220 patients, patients with poor outcome were found to satisfy 2 rules, i.e., high expression of MAX dimerization with high expression of unknown A (LC_26146), or high expression of MAX dimerization with low expression of unknown B (LC_33144). The present paper is the first to describe the multiple noninferior FNN modeling system. This system is a powerful tool for predicting outcome and classifying patients, and is applicable to other heterogeneous diseases. PMID- 14556666 TI - Helicobacter pylori risk associated with childhood home environment. AB - Helicobacter pylori (H. pylori) is considered to be a cause of gastric ulcer, gastric cancer and other diseases. The relationship between infection and the hygiene or housing circumstances of such patients in their childhood was explored. The study subjects were those who applied for a H. pylori antibody test, and were asked to fill out a questionnaire enclosed with a test kit, inquiring as to their hygiene and housing conditions when they were 10 years old. Of 5971 applicants, 5854 agreed to participate in the study. Associations between the six factors in the questionnaire and infection were calculated, and adjusted for sex, age and district. Drinking water, type of toilet, residential area, number of people in the house, and birth order showed significant correlations with H. pylori infection. The odds ratios (95% confidence intervals) were 0.73 (0.55-0.96) for tap water, 0.72 (0.63-0.84) for flush toilets, 0.74 (0.66-0.83) for urban location, 1.34 (1.09-1.64) for 7 or more people in the household, 1.19 (1.00-1.41) for 4th or 5th in birth order, and 1.47 (1.17-1.85) for 6th or more in birth order. No significant association with breast feeding was observed. These results suggest that infection with H. pylori may be associated with water related sanitary factors in childhood, and that the bacillus may be transmitted within a family. PMID- 14556667 TI - Projection of lung cancer mortality in Japan. AB - According to the National Vital Statistics data, age-standardized mortality rates (ASRs) of lung cancer have shown slightly declining trends in Japan for both men and women. In order to evaluate whether this tendency will continue, a Bayesian age-period-cohort (APC) model was applied using the National Vital Statistics data from 1952 to 2001. In the projection, a Gaussian autoregressive prior model was applied to smooth age, period, and cohort effects from its 2 immediate predecessors by extrapolation. Posterior distributions from which we drew inferences on mortality rates were derived from 15,000 iterations using 5000 burn in iterations. We defined the median of the iterated values as the overall summary mortality rate of the iterated results. Our results suggest that the number of deaths due to lung cancer will double for men and women during the next 3 decades due to the aging of the baby-boomer generation (individuals who were born between 1947 and 1951). Currently declining trends in some age groups will reverse and start to increase again in the next decades. However, for recent birth cohorts, the results of the projection varied according to whether the data set included early age group mortality or not. Lung cancer mortality in the future depends on the risk factors engaged in by today's young people, especially smoking. Strong promotion of anti-smoking measures and careful surveillance for lung cancer are needed. PMID- 14556668 TI - Critical role of the Th1/Tc1 circuit for the generation of tumor-specific CTL during tumor eradication in vivo by Th1-cell therapy. AB - Th1 and Th2 cells obtained from OVA-specific T cell receptor transgenic mice completely eradicated the tumor mass when transferred into mice bearing A20-OVA tumor cells expressing OVA as a model tumor antigen. To elucidate the role of Tc1 or Tc2 cells during tumor eradication by Th1- or Th2-cell therapy, spleen cells obtained from mice cured of tumor by the therapy were re-stimulated with the model tumor antigen (OVA) for 4 days. Spleen cells obtained from mice cured by Th1-cell therapy produced high levels of IFN-gamma, while spleen cells from mice cured by Th2-cell therapy produced high levels of IL-4. Intracellular staining analysis demonstrated that a high frequency of IFN-gamma-producing Tc1 cells was induced in mice given Th1-cell therapy. In contrast, IL-4-producing Tc2 cells were mainly induced in mice after Th2-cell therapy. Moreover, Tc1, but not Tc2, exhibited a tumor-specific cytotoxicity against A20-OVA but not against CMS-7 fibrosarcoma. Thus, immunological memory essential for CTL generation was induced by the Th1/Tc1 circuit, but not by the Th2/Tc2 circuit. We also demonstrated that Th1-cell therapy is greatly augmented by combination therapy with cyclophosphamide treatment. This finding indicated that adoptive chemoimmunotherapy using Th1 cells should be applicable as a novel tool to enhance the Th1/Tc1 circuit, which is beneficial for inducing tumor eradication in vivo. PMID- 14556670 TI - Overview of antibody therapy in B-cell non-Hodgkin's lymphoma. AB - The main treatment modalities for lymphoma in the past decade have been radiation therapy and chemotherapy. Recently, molecular engineering provided humanized antibodies with promising clinical activity, and rituximab is the first commercially available antibody. This anti-CD20 monoclonal antibody (MoAb) showed little toxicity and demonstrated excellent clinical activity. Given as a single agent, it induces a high-response rate even in pretreated low-grade non-Hodgkin's lymphoma, the effect being higher if administered for a prolonged period of time. Its action is synergistic with chemotherapy, and combination treatment could improve survival in patients with aggressive lymphomas. Rituximab also demonstrated the ability to clear tumor cells from the circulation, allowing for an in vivo purging effect in the setting of peripheral stem cell collection and transplantation. Still, a number of issues related to its use need to be addressed, such as optimal dose and schedule and the situations in which rituximab should be given as a single agent or in addition to chemotherapy or other drugs, such as other MoAbs or interferons. We also need to understand when rituximab should be used in first-line treatment, with which type of chemotherapy the combination is most cost-effective, and patient population that will benefit most from this antibody treatment. PMID- 14556671 TI - Epoetin alfa therapy for patients with hematologic malignancies and mild anemia. AB - Anemia has been reported in approximately 40%-70% of patients with hematologic malignancies, with severity depending on the type and stage of disease and whether the patient has received myelosuppressive chemotherapy. Growing evidence supports the role of epoetin alfa in correcting anemia and improving quality of life (QOL) in patients with hematologic malignancies. Clinical practice guidelines recommend the use of epoetin alfa in patients with cancer-related anemia (including patients with hematologic malignancies) and hemoglobin levels < or =10 g/dL. Epoetin alfa treatment is optional for patients with cancer-related anemia and hemoglobin levels>10 g/dL and <12 g/dL, depending on clinical circumstances. A prospective, open-label, randomized trial evaluating hematologic response, transfusion use, and QOL after immediate or delayed epoetin alfa treatment in mildly anemic patients (hemoglobin< or =12 g/dL) undergoing chemotherapy for chronic lymphocytic leukemia, multiple myeloma, or lymphoma was recently completed. Study objectives included determining any correlation between changes in hemoglobin level and QOL and assessing any correlation between QOL measures and health care resource use. Interim results suggest that epoetin alfa treatment in patients with hematologic cancers and hemoglobin< or =12 g/dL who are receiving chemotherapy increases hemoglobin, functional capacity, well-being, work and productivity, and health resource use. Further evaluation of alternative epoetin alfa dosing schedules and use of epoetin alfa in treating anemia in patients with specific hematologic malignancies is ongoing. PMID- 14556672 TI - Pegylated liposomal doxorubicin, vincristine, and reduced-dose dexamethasone as first-line therapy for multiple myeloma. AB - In patients with multiple myeloma (MM) who may ultimately receive active therapy, the combination of VAD (vincristine, doxorubicin, and dexamethasone) has been shown to be effective. However, the use of VAD is complicated by inherent risks that result from the use of central venous catheters, steroid toxicity, and by doxorubicin-associated adverse events such as cardiotoxicity and alopecia. To address these issues, a phase II trial investigating the combination of vincristine, pegylated liposomal doxorubicin, and reduced-schedule oral dexamethasone in the first-line treatment of patients with MM has been conducted. Patients with symptomatic, newly diagnosed MM were treated with intravenous (i.v.) pegylated liposomal doxorubicin 40 mg/m2 and vincristine 2 mg on day 1, along with dexamethasone 40 mg/day given either i.v. or orally for 4 days, every 4 weeks for a minimum of 6 cycles. Responses were reported in 29 patients (88%), and an additional 3 patients achieved stable disease. The median time to maximal response was 5.8 months (range, 0.7-13.6 months), and median overall survival time is estimated to be 60 months. This treatment regimen was well tolerated, and the most common grade 3/4 adverse events included hand-foot syndrome (21%), neutropenia (30%), anemia (21%), and mucositis (12%). Based on these results, the vincristine/liposomal doxorubicin/dexamethasone regimen appears to be effective and well tolerated in the first-line treatment of MM. PMID- 14556674 TI - The role of erythropoietin in the anemia of myelodysplastic syndrome. AB - Although erythropoietin (EPO) deficiency is not responsible for the anemia of myelodysplasia, pharmacologic doses of recombinant human EPO (rHuEPO, epoetin alfa) and epoetin beta have been studied extensively as treatment of anemia in myelodysplastic syndrome (MDS). When an epoetin is used as a single growth factor in patients with MDS, clinically meaningful responses occur in only a small minority of patients (16%). Patients who are transfusion-dependent are less likely to respond than patients who are transfusion-independent. Serum EPO level has a weak association with response rate and cannot be used to select or exclude patients from empirical trials of epoetins. The dose schedule commonly used as initial treatment 40,000 U/week, is consistent with clinical observations, but an optimal dose schedule has not been determined. The combination of an epoetin and granulocyte colony-stimulating factor (G-CSF) produces a higher erythroid response rate (36%) than the regimen of epoetin alone, but we have found no randomized trial data to support this point. However, the design of the clinical trials, which included adding G-CSF after epoetin alone had failed, supports the hypothesis that combined use of growth factors, rather than just higher doses of epoetin, is responsible for the high response rate observed with the combination of epoetin and G-CSF. Unfortunately, as in the case of epoetin alone, patients who are transfusion-dependent (> or =2 U red blood cells/month) are less likely to respond to combined growth factor therapy. Although the ability of patients with MDS to show an erythroid response to epoetin is of biologic interest, because of high costs and the limited response rate in transfusion-dependent patients, epoetin therapy, with or without G-CSF, cannot be regarded as a definitive therapy for the anemia of MDS. PMID- 14556673 TI - Farnesyltransferase inhibitors in acute myeloid leukemia and myelodysplastic syndromes. AB - Farnesyltransferase inhibitors were initially developed as Ras inhibitors as they inhibit the prenylation necessary for Ras activation. It is clear now that their mechanism of action is more complex and probably involves other proteins unrelated to Ras. At least 3 drugs within this family have been investigated in acute myeloid leukemia, myelodysplastic syndromes, and other leukemias. These are tipifarnib (R115777, Zarnestra), lonafarnib (SCH66336, Sarasar), and BMS-214662. The first 2 are administered orally, whereas BMS-214662 is given intravenously. These drugs are at different stages of development, and design of treatment schedules and methodology of the available studies are very different. Although most of the information is still preliminary, these agents have demonstrated clear evidence of clinical activity in these diseases and very favorable toxicity profiles. Several studies are still ongoing to better define the efficacy of these agents in the treatment of leukemias, as well as to determine the best schedules, the role of combination with other agents, and the role of these agents in different settings, such as the management of minimal residual disease. It is very possible that these agents will soon find their way to the ranks of established agents for the management of myeloid malignancies PMID- 14556676 TI - 39th annual meeting of the American Society of Clinical Oncology. PMID- 14556675 TI - The implications of anemia in multiple myeloma. AB - Multiple myeloma (MM) is commonly associated with anemia. Several causes have been implicated, but anemia of chronic disease with inadequate erythropoietin (EPO) production related to the inflammatory cytokines appears to be of utmost importance. Interleukin-1 and tumor necrosis factor are capable of suppressing erythropoiesis. Anemia has broad implications. First, the low hemoglobin and hematocrit are associated with poor quality of life and performance and affect daily activity. Second, anemia has an impact on the cardiovascular system. Considering that most MM patients are elderly, this may be even more important. Anemia has been shown to induce or aggravate hypoxia and ischemic complications. Third, anemia has been shown to be a poor prognostic factor in MM. Traditionally, patients with symptomatic anemia were treated with red blood cell transfusions as needed. The introduction of epoetin alfa and epoetin beta into clinical practice opened new avenues to these patients. The administration of epoetins to patients with MM and anemia have been shown to be very useful. Several studies in more than 1000 patients have demonstrated a high response rate (range, 25%-85%; mean, 60%). This response is characterized by a significant increase of hemoglobin, hematocrit, and the number of red blood cells together with a reduction in the blood transfusion requirements. This is also associated with an improved quality of life. Although there is no complete agreement about the role of pretreatment serum EPO levels, many investigators believe that relatively low levels may help in predicting response, thereby limiting the number of potential candidates to receive this expensive therapy. The epoetins are safe and well tolerated with minimal toxicity; however, some concern has been recently raised regarding several dozen patients who developed pure red cell aplasia while on epoetin therapy. However, this adverse effect appears to be extremely rare. Recent data suggest that EPO has additional biologic effects, such as longer-than-expected survival in patients with MM. This observation is further supported by animal studies, demonstrating an antimyeloma effect of EPO in mice models. This effect has been shown to be immune mediated. If these exciting data are confirmed in future clinical trials, this may have significant implications on the treatment of MM. PMID- 14556679 TI - Acquired immunodeficiency syndrome-related lymphoma in the era of highly active antiretroviral therapy. AB - The treatment and outcome of human immunodeficiency virus (HIV) infection altered dramatically in the mid-1990s with the introduction of highly active antiretroviral therapy (HAART). Highly active antiretroviral therapy, where available, has led to a dramatic decline in mortality from HIV and a decrease in the incidence of opportunistic infections and Kaposi sarcoma. This article addresses the effects that HAART has had on acquired immunodeficiency syndrome (AIDS)-related non-Hodgkin's lymphoma (NHL). Metaanalysis of numerous cohort studies confirmed that the incidence of AIDS-related NHL has decreased since the advent of HAART. This decline is most marked for primary cerebral lymphomas and systemic immunoblastic lymphoma but has not been demonstrated for Burkitt lymphoma. In addition to genetic predisposing factors, age, nadir CD4 cell count, and lack of HAART therapy predict the development of NHL. The clinical presentation of AIDS-related NHL has not changed, but several institutions have reported an improvement in survival since the introduction of HAART. Moreover, HAART has been combined safely with systemic chemotherapy in the management of NHL, and this approach results in a more modest decrease in immune function than when chemotherapy is administered alone. This has led to a more aggressive approach to the management of AIDS-related NHL and response rates and overall survival durations that are approaching those seen in stage-matched high-grade lymphomas in the immunocompetent population. PMID- 14556680 TI - Fourteen-day CHOP supported with granulocyte colony-stimulating factor in patients with aggressive non-Hodgkin's lymphoma: results of a phase II study. AB - The safety and efficacy of compressed-cycle (14-day) standard-dose CHOP (cyclophosphamide/doxorubicin/vincristine/prednisone) supported with prophylactic recombinant granulocyte colony-stimulating factor (G-CSF; filgrastim) were evaluated in patients with aggressive non-Hodgkin's lymphoma (NHL). Patients with intermediate- or high-grade NHL (Working Formulation groups D-H and J; N = 120), accrued from 25 clinical practices, were given 6 cycles of standard-dose CHOP every 14 days. Granulocyte colony-stimulating factor 5 microg/kg was given daily subcutaneously in each cycle, starting on day 2 and continuing until the absolute neutrophil count was = 10000/microL. The overall response rate was 89%, with complete responses (CRs) in 52 of 120 patients (43%) and partial responses in 55 (46%). These results are consistent with previously reported outcomes from trials in this population. Of the 720 chemotherapy cycles planned for all patients, 615 (85%) were given on time at full dose. The median relative dose intensity (RDI) of cyclophosphamide and doxorubicin was 99%; the RDI of vincristine was 73%. In the 53 patients = 60 years of age, 80% of the chemotherapy cycles were given on time at full dose, with median RDIs similar to those in the entire population. Response rates in the older patients were also similar, with CRs in 24 patients (45%) and partial responses in 21 (40%). Hematologic toxicity was significant but tolerable, with no treatment-related deaths. At a median follow-up of 20.6 months, 77% of patients were still alive. Standard-dose CHOP administered every 14 days with prophylactic G-CSF support was delivered as planned in most patients and produced response rates comparable with those with CHOP given every 3 weeks, without exceptional toxicity. PMID- 14556681 TI - Primary gastrointestinal non-Hodgkin's lymphoma: treatment outcome. AB - Primary gastrointestinal lymphoma is the most common extranodal presentation of non-Hodgkin's lymphoma. In this retrospective study, all cases of primary gastrointestinal lymphoma registered at Kuwait Cancer Control Center between January 1992 and December 2001 were reviewed. Patients were diagnosed through an endoscopic biopsy or exploratory laparotomy. Staging procedures included computed axial tomography scan, bone marrow biopsy, barium and follow-through study, as well as routine chemistry (including lactate dehydrogenase) and hemography. There were 46 patients (30 men and 16 women); ages ranged from 16 to 82 years (median age, 43.6 years). The most common site of involvement was the stomach (70%), and the most common histology was large B cell. At least 50% of cases underwent primary radical surgery, and the majority of patients received subsequent chemotherapy and/or local radiation therapy. Chemotherapy consisted of the CHOP (cyclophosphamide/doxorubicin/vincristine/prednisone) regimen for large B-cell histology and CVP (cyclophosphamide/vincristine/prednisone) or oral alkylating agents for low-grade histology. Of this cohort, only 12 cases (26%) have had local or distant relapse. Disease-free survival rates for low-grade and high grade non-Hodgkin's lymphoma were 64% and 81%, respectively. Primary gastrointestinal lymphoma is a potentially curable malignancy. PMID- 14556682 TI - Lymphomas and oncolytic virus therapy. AB - There are several well-documented cases in medical literature of the remission of leukemias and malignant lymphomas following natural human viral infections. In the hope of being able to reproduce these spontaneous tumor regressions, investigators have studied various viruses with distinct oncolytic properties. The first attempts to treat patients with oncolytic viruses took place > 80 years ago; however, it achieved little success. With modern technologies and current knowledge of viruses and cancer, there is an expectation for the discovery of efficient oncolytic viral therapies. This article will review the current knowledge of oncolytic viruses in relation to the treatment of lymphoma. PMID- 14556683 TI - Multimodality treatment in a case of primary cardiac lymphoma. AB - Primary cardiac lymphoma(PCL) is an exceptionally rare entity associated with a poor progonosis. The patient in this report underwent successful surgical resection of a PCL. We now describe her multimodality treatment including autologous stem cell transplantation which resulted in a 22 month survival. PMID- 14556684 TI - Rituximab in lymphocyte predominance Hodgkin's disease: a case series. AB - Rituximab in combination with chlorambucil or radiation therapy may be an effective and less-toxic therapeutic alternative for patients with lymphocyte predominance Hodgkin's disease (LPHD). We treated 6 patients with LPHD with weekly rituximab at 375 mg/m2 for 4 weeks, followed by either radiation therapy or chlorambucil. Four patients had previously untreated disease and 2 had relapsed LPHD. All patients had no evidence of disease progression at a median follow-up time of 12.5 months after receiving rituximab therapy (range, 6-39 months) and a median follow-up time of 6.5 months after completion of chlorambucil or radiation therapy (range, 3-25 months). Further follow-up is warranted to evaluate response duration and late toxicity of this novel treatment strategy PMID- 14556685 TI - A prospective, open-label safety and efficacy study of combination treatment with bortezomib (PS-341, velcade and melphalan in patients with relapsed or refractory multiple myeloma. PMID- 14556686 TI - Artificial fish schools: collective effects of school size, body size, and body form. AB - Individual-based models of schooling in fish have demonstrated that, via processes of self-organization, artificial fish may school in the absence of a leader or external stimuli, using local information only. We study for the first time how body size and body form of artificial fish affect school formation in such a model. For a variety of group sizes we describe how school characteristics (i.e., group form, spread, density, polarization, turning rate, and speed) depend on body characteristics. Furthermore, we demonstrate that the nearest neighbor distance and turning rate of individuals are different for different regions in the group, although the agents are completely identical. Our approach shows the significance of both self-organization and embodiment in modeling of schools of artificial fish and, probably, in structuring schools of real fish. PMID- 14556687 TI - Evolving mobile robots able to display collective behaviors. AB - We present a set of experiments in which simulated robots are evolved for the ability to aggregate and move together toward a light target. By developing and using quantitative indexes that capture the structural properties of the emerged formations, we show that evolved individuals display interesting behavioral patterns in which groups of robots act as a single unit. Moreover, evolved groups of robots with identical controllers display primitive forms of situated specialization and play different behavioral functions within the group according to the circumstances. Overall, the results presented in the article demonstrate that evolutionary techniques, by exploiting the self-organizing behavioral properties that emerge from the interactions between the robots and between the robots and the environment, are a powerful method for synthesizing collective behavior. PMID- 14556688 TI - Bridging nonliving and living matter. AB - Assembling non-biological materials (geomaterials) into a proto-organism constitutes a bridge between nonliving and living matter. In this article we present a simple step-by-step route to assemble a proto-organism. Many pictures have been proposed to describe this transition within the origins-of-life and artificial life communities, and more recently alternative pictures have been emerging from advances in nanoscience and biotechnology. The proposed proto organism lends itself to both traditions and defines a new picture based on a simple idea: Given a set of required functionalities, minimize the physicochemical structures that support these functionalities, and make sure that all structures self-assemble and mutually enhance each other's existence. The result is the first concrete, rational design of a simple physicochemical system that integrates the key functionalities in a thermodynamically favorable manner as a lipid aggregate integrates proto-genes and a proto-metabolism. Under external pumping of free energy, the metabolic processes produce the required building blocks, and only specific gene sequences enhance the metabolic kinetics sufficiently for the whole system to survive. We propose an experimental implementation of the proto-organism with a discussion of our experimental results, together with relevant results produced by other experimental groups, and we specify what is still missing experimentally. Identifying the missing steps is just as important as providing the road map for the transition. We derive the kinetic and thermodynamic conditions of each of the proto-organism subsystems together with relevant theoretical and computational results about these subsystems. We present and discuss detailed 3D simulations of the lipid aggregation processes. From the reaction kinetics we derive analytical aggregate size distributions, and derive key properties of the metabolic efficiency and stability. Thermodynamics and kinetics of the ligation directed self-replication of the proto-genes is discussed, and we summarize the full life cycle of the proto-organism by comparing size, replication time, and energy with the biomass efficiency of contemporary unicells. Finally, we also compare our proto-organism picture with existing origins-of-life and protocell pictures. By assembling one possible bridge between nonliving and living matter we hope to provide a piece in the ancient puzzle about who we are and where we come from. PMID- 14556689 TI - Selective instability: maternal effort and the evolution of gene activation and deactivation rates. AB - We previously used simulations of gene expression to demonstrate that rapid activation and deactivation rates stabilized outcomes in stochastic systems. We hypothesized that transient single allele inactivation of an autosomal gene during gametogenesis or very early embryogenesis could have a selective advantage if it permits the functional sampling of each allele and precludes committing maternal effort to an embryo with a deleterious mutation. To test this hypothesis, we simulated the evolution of gene expression activation and deactivation rates and imposed two different selective pressures on the populations: (a). late selection against individuals that cannot maintain a threshold level of gene product that occurs after the investment of maternal effort (i.e., after birth); or (b). early selection: in addition to late selection, maintenance of the gene product above a threshold level was necessary for early development prior to commitment of maternal effort. We found that the opportunity to save reproductive effort from early selection caused the evolution of higher deactivation rates and lower activation rates than in the late selection condition. Thus, we predict that in the special case where early selection can save maternal investment in non-viable offspring, gene expression activation rates and deactivation rates might be selected to permit sampling of the product from each allele. PMID- 14556690 TI - Viscous populations and their support for reciprocal cooperation. AB - Viscous populations (those whose members are spatially distributed and have limited mobility and locality of interaction and mating) have been proposed to support the evolution of reciprocal cooperation among self-interested individuals. Here we present a model of such a population and describe how its examination yielded the realization that different classes of viscous populations exist with differing levels of support for reciprocal cooperation. Specifically we find from our model that, in a spatially distributed population with increased viscosity, the reciprocally cooperative tit-for-tat strategy may not be globally stable due to a corresponding increase in local population density. PMID- 14556691 TI - Production effects related to mastitis and mastitis economics in dairy cattle herds. AB - Mastitis is the most prevalent production disease in dairy herds world-wide and is responsible for several production effects. Milk yield and composition can be affected by a more or less severe short-term depression and, in case of no cure, by a long-acting effect, and, sometimes, an overlapping effect to the next lactation. Summary values in the literature for losses of milk production were proposed at 375 kg for a clinical case (5% at the lactation level) and at 0.5 kg per 2-fold increase of crude SCC of a cow. Due to the withdrawal period after treatment, composition changes in milk can almost be neglected in economic calculations. Lethality rate for clinical mastitis is very low on the average, while anticipated culling occurs more frequently after clinical and subclinical mastitis (relative risk between 1.5 and 5.0). The economics of mastitis needs to be addressed at the farm level and, per se, depends on local and regional epidemiological, managerial and economic conditions. To assess the direct economic impact of mastitis, costs (i.e. extra resource use) and losses (i.e. reduced revenues) have to be aggregated. To support decision making for udder health control, it is necessary to use a marginal approach, based on the comparison of the losses avoided and the additional costs of modified plans, compared to the existing ones. PMID- 14556692 TI - Statistical modelling for clinical mastitis in the dairy cow: problems and solutions. AB - Modelling case occurrence and risk factors for clinical mastitis, as a key multifactorial disease in the dairy cow, requires statistical models. The type of model used depends on the choice of perception or the study level: herd, lactation, animal, udder and quarter. The validity of the tests that are performed through these models is especially ensured when hypotheses of independence between statistical units are respected, and when the model adjustments do not involve overdispersion faced with the observed data. In the article, the main sources of overdispersion are identified according to the different levels of perception of mastitis risk. Then, the proposed solutions to control for overdispersion at each study level are discussed and the difficulty to compare the study results is highlighted through a variety of methodological choices of the authors. Two main categories of models are used for modelling clinical mastitis, i.e. generalist exploratory models and explanatory designed models. The contribution of the explanatory models to improve modelling accuracy and relevance is documented through the two main published methodological approaches, the first one being based on a states model, and the second on a survival model. The integration and optimisation of such explanatory modelling methods should be possible in the future in order to develop a more global explanatory model including herd risk factors, which could pertinently predict udder infections (both clinical and subclinical) at the cow, lactation, or even udder and quarter levels. PMID- 14556693 TI - Coliform mastitis. AB - Gram-negative bacteria that commonly cause bovine mastitis are classified as environmental pathogens. The point sources of coliform bacteria that cause infections include bedding materials, soil, manure and other organic matter in the environment of cows. Rates of coliform mastitis increase during climatic periods that maximize populations in the environment. The portal of entry into the mammary gland for Gram-negative bacteria is the teat canal. Once in the gland, bacteria must utilize available substrates in the mammary secretion to replicate and evade host defenses. Rates of coliform mastitis are greater during the transitional phases of the non- lactating period than during lactation. The ability to infect the non-lactating gland is directly related to the ability of bacteria to acquire iron from the mammary secretion. The primary host defense against coliform mastitis during lactation is the elimination of bacteria by neutrophils migrating into the gland in response to inflammation. Damage to the host is mediated by the release of endotoxin. The severity and duration of clinical signs associated with coliform mastitis are reduced by the use of core antigen bacterins. PMID- 14556694 TI - Severity of E. coli mastitis is mainly determined by cow factors. AB - Intramammary infections of dairy cows with Gram-positive bacteria such as Staphylococcus aureus (major cause of mastitis) have received a lot of attention because of their major economic impact on the dairy farm through production losses induced by an increase in somatic cell count. Management strategies, including greater awareness for efficient milking and hygienic measures, have limited the spread of Gram-positive bacteria and resulted in a significant decrease of proportion of S. aureus isolates and subclinical mastitis worldwide. Other organisms such as coliform subspecies and Streptococcus uberis, both environmental bacteria that cause clinical mastitis, have received less attention. Escherichia coli causes inflammation of the mammary gland in dairy cows around parturition and during early lactation with striking local and sometimes severe systemic clinical symptoms. This disease affects many high producing cows in dairy herds and may cause several cases of death per year in the most severe cases. It is well known that bacterial, cow and environmental factors are interdependent and influence mastitis susceptibility. Many studies, executed during the last decade, indicate that the severity of E. coli mastitis is mainly determined by cow factors rather than by E. coli pathogenicity. During E. coli mastitis, the host defense status is a cardinal factor determining the outcome of the disease. Today, we know that the neutrophil is a key factor in the cows' defense against intramammary infection with E. coli. Effective elimination of the pathogen by neutrophils is important for the resolution of infection and the outcome of E. coli mastitis. This review is a compilation of some major findings over the last 15 years concerning mainly host factors that modulate and influence neutrophil function and the mammary inflammatory reaction. The individual chapters address: virulence factors of E. coli strains, how neutrophils kill E. coli, connection between endotoxins, tumor necrosis factor alpha and nitric oxide, severity classification of E. coli mastitis, lifespan of neutrophils, host factors that influence severity, tissue damage and production loss. PMID- 14556695 TI - Indicators of inflammation in the diagnosis of mastitis. AB - Mastitis affects the quality of milk and is a potential health risk for the other cows. In a well managed dairy herd, in addition to clinical mastitis, subclinical mastitis should be efficiently detected. Bacteriological sampling is not feasible as a routine test to identify subclinical mastitis, and indirect tests of mastitis are more suitable for selecting cows with intramammary infections for subsequent bacteriological sampling. Mastitis affects the composition of milk, and the degree of changes depends on the infecting agent and the inflammatory response. Indicators of inflammation in the milk which can be determined using rapid, reliable and easy routine techniques, can be used for the early detection of mastitis. The measuring of the somatic cell count in milk is the standard method, but the analysis technique is problematic for routine use in herds. The most promising parameters for monitoring subclinical mastitis are milk N-acetyl beta-D-glucosaminidase activity, lactose, and electrical conductivity along with some other indicators such as optical and milk flow measurements, preferably with an inter-quarter evaluation included in the test. Acute phase proteins, haptoglobin and serum amyloid A, are also potential candidates for mastitis monitoring. New mastitis detection systems which can be adapted into on-line use are urgently needed, since dairy units are growing bigger and automatic milking systems are being taken into use. PMID- 14556696 TI - Monitoring udder health and milk quality using somatic cell counts. AB - In this article the use of somatic cell counts for monitoring udder health and milk quality is discussed. Somatic cell count dynamics at quarter, cow, herd and population level are discussed and illustrated with examples. Quarter and cow somatic cell counts directly represent the inflammatory status of the mammary gland. Herd and population somatic cell count are related to the inflammatory process in individual cows but much more reflect the udder health status of the herd and the quality of the raw milk in the herd and the population. Application of monitoring tools in herd health management are illustrated using a case study. Understanding infection dynamics requires precise longitudinal data. Monitoring tools are required to find the areas of risk in the herd. It is inevitable that more complete udder health programs and monitoring systems are to be developed and implemented. These programs are necessarily dynamic and complex. Implementation of complete udder health programs should be accompanied by research efforts to further fine-tune these complete udder health control and monitoring programs. PMID- 14556697 TI - The bovine neutrophil: Structure and function in blood and milk. AB - Migration of polymorphonuclear neutrophil leukocytes (PMN) into the mammary gland provide the first line of defense against invading mastitis pathogens. Bacteria release potent toxins that activate white blood cells and epithelial cells in the mammary gland to secrete cytokines that recruit PMN that function as phagocytes at the site of infection. While freshly migrated PMN are active phagocytes, continued exposure of PMN to inhibitory factors in milk such as fat globules and casein, leads to altered PMN morphology and reduced phagocytosis. In the course of phagocytosing and destroying invading pathogens, PMN release chemicals that not only kill the pathogens but that also cause injury to the delicate lining of the mammary gland. This will result in permanent scarring and reduced numbers of milk secretory cells. The life span of PMN is limited by the onset of apoptosis. To minimize damage to mammary tissue, PMN undergo a specialized process of programmed cell death known as apoptosis. Macrophages quickly engulf and phagocytose apoptotic PMN, thereby minimizing the release of PMN granular contents that are damaging to tissue. The PMN possess an array of cell surface receptors that allow them to adhere and migrate through endothelium and to recognize and phagocytose bacteria. One receptor found on phagocytes that is receiving considerable attention in the control of infections by Gram-negative bacteria is CD14. Binding of lipopolysaccharide (LPS) to membrane bound CD14 causes release of tumor necrosis factor-alpha and sepsis. Binding of LPS to soluble CD14 shed from CD14-bearing cells results in neutralization of LPS and rapid recruitment of PMN to the site of infection. Recent advances in the fields of genomics and proteomics should greatly enhance our understanding of the PMN role in controlling intramammary infections in ruminants. Further, manipulation of PMN, through either recombinant proteins such as soluble CD14 that enhance PMN response or agents that mediate PMN apoptosis, may serve as novel therapeutics for the treatment of mastitis. PMID- 14556698 TI - Polymorphonuclear proteolytic activity and milk composition change. AB - Relationships between Quarter Milk Cell Count (QMCC) and Tank Milk Cell Count (TMCC) with milk biochemical and technological parameters in milk and dairy products were investigated. All parameters measured were affected by the increase of TMCC and QMCC between 0 and 600 000 cells/mL. The variable effect of lactation stage which is different for different authors, is discussed. The three mechanisms, measured during the inflammation of the udder, implicated in the modification of milk quality are described (a decrease in synthesis, a decrease in the milk barrier permeability and an increase in proteolytic activities). The direct effect of plasmin in caseinolysis is well known; the specific role of the increase of somatic cells (especially PMN) in the modification of milk quality is described. Several specific proteolytic activities of PMN are described and the impact of these activities on caseinolysis is evaluated. Two hypothetical mechanisms of caseinolysis by PMN are suggested and a synthetic scheme of the role of plasmin, bacteria and somatic cells in caseinolysis is discussed. PMID- 14556699 TI - The complement in milk and defense of the bovine mammary gland against infections. AB - The mammary gland of dairy cows, which is prone to infection by various bacteria, mobilizes local and systemic immune defenses to cope with pathogens. The complement system plays an important part in the innate immunity against microorganisms through its bactericidal, opsonic, and phlogistic functions. The amount of the complement in the milk of healthy glands of dairy cows is low. Moreover, the classical pathway of activation is not functional because of a shortage in C1q. By contrast, the alternative pathway is active, deposits C3b and C3bi on bacteria, and generates amounts of C5a which are highly variable among cows. A slight inhibition of the bactericidal/hemolytic activities, of the deposition of C1q on bacteria, and of the phlogistic activity of C5a makes milk a rather anti-inflammatory fluid. The inhibitory activity does not involve C3b/ C3bi deposition on bacteria, nor the generation of C5a by the alternative pathway. When inflammation develops, the blood-derived complement components overcome the inhibitions and complement-dependent bactericidal, opsonic and phlogistic activities may be high in milk. Further research is necessary to evaluate the contribution of C5a to the recruitment of leukocytes in the mammary gland, and to specify the links between the complement system and the response of resident cells (leukocytes and mammary epithelial cells) to infection stimulus. This will help to define the contribution of the complement system to resistance against mastitis, and could help to differentiate animals more or less resistant to this frequent and costly disease. PMID- 14556700 TI - Genetics of resistance to mastitis in dairy cattle. AB - Genetic variability of mastitis resistance is well established in dairy cattle. Many studies focused on polygenic variation of the trait, by estimating heritabilities and genetic correlation among phenotypic traits related to mastitis such as somatic cell counts and clinical cases. The role of Major Histocompatibility Complex in the susceptibility or resistance to intrammamary infection is also well documented. Finally, development from molecular genome mapping led to accumulating information of quantitative trait loci (QTL) related to mastitis resistance and better understanding of the genetic determinism of the trait. From economic and genetic analyses, and according to welfare and food safety considerations and to breeders and consumer's concern, there is more and more evidence that mastitis should be included in breeding objective of dairy cattle breeds. Many countries have implemented selection for mastitis resistance based on linear decrease of somatic cell counts. Given biological questioning, potential unfavourable consequences for very low cell counts cows are regularly investigated. Improvement of selection accuracy for mastitis resistance is ongoing and includes: advances in modelling, optimal combination of mastitis related traits and associated predictors, such as udder morphology, definition of global breeding objective including production and functional traits, and inclusion of molecular information that is now available from QTL experiments. PMID- 14556701 TI - Mastitis of dairy small ruminants. AB - Staphylococci are the main aetiological agents of small ruminants intramammary infections (IMI), the more frequent isolates being S. aureus in clinical cases and coagulase negative species in subclinical IMI. The clinical IMI, whose annual incidence is usually lower than 5%, mainly occur at the beginning of machine milking and during the first third of lactation. These features constitute small ruminant peculiarities compared to dairy cattle. Small ruminant mastitis is generally a chronic and contagious infection: the primary sources are mammary and cutaneous carriages, and spreading mainly occurs during milking. Somatic cell counts (SCC) represent a valuable tool for prevalence assessment and screening, but predictive values are better in ewes than in goats. Prevention is most often based on milking machine management, sanitation and annual control, and milking technique optimisation. Elimination mainly relies on culling animals exhibiting clinical, chronic and recurrent IMI, and on drying-off intramammary antibiotherapy; this treatment allows a good efficacy and may be used selectively by targeting infected udders only. Heritability values for lactation mean SCC scores are between 0.11 and 0.15. Effective inclusion of ewe's mastitis resistance in the breeding goal has recently been implemented in France following experimental and large scale estimations of genetic parameters for SCC scores. PMID- 14556703 TI - 15 years of neuron cell biology. PMID- 14556704 TI - Neurogenesis and the cell cycle. AB - For a long time, it has been understood that neurogenesis is linked to proliferation and thus to the cell cycle. Recently, the gears that mediate this linkage have become accessible to molecular investigation. This review describes some of the progress that has been made in understanding how the molecular machinery of the cell cycle is used in the processes of size regulation in the brain, histogenesis, neuronal differentiation, and the maintenance of stem cells. PMID- 14556705 TI - Cytoskeletal dynamics and transport in growth cone motility and axon guidance. AB - Recent studies indicate the actin and microtubule cytoskeletons are a final common target of many signaling cascades that influence the developing neuron. Regulation of polymer dynamics and transport are crucial for the proper growth cone motility. This review addresses how actin filaments, microtubules, and their associated proteins play crucial roles in growth cone motility, axon outgrowth, and guidance. We present a working model for cytoskeletal regulation of directed axon outgrowth. An important goal for the future will be to understand the coordinated response of the cytoskeleton to signaling cascades induced by guidance receptor activation. PMID- 14556706 TI - The control of dendrite development. AB - Dendrite development is an important and unsolved problem in neuroscience. The nervous system is composed of a vast number of neurons with strikingly different morphology. Neurons are highly polarized cells with distinct subcellular compartments, including one or multiple dendritic processes arising from the cell body, and a single, extended axon. Communications between neurons involve synapses formed between axons of the presynaptic neurons and dendrites of the postsynaptic neurons. Extensive studies over the past decade have identified many molecules underlying axonal outgrowth and pathfinding. In contrast, the control of dendrite development is still much less well understood. However, recent progress has begun to shed light on the molecular mechanisms that orchestrate dendrite growth, arborization, and guidance. PMID- 14556707 TI - Mechanisms of synapse assembly and disassembly. AB - The mechanisms that govern synapse formation and elimination are fundamental to our understanding of neural development and plasticity. The wiring of neural circuitry requires that vast numbers of synapses be formed in a relatively short time. The subsequent refinement of neural circuitry involves the formation of additional synapses coincident with the disassembly of previously functional synapses. There is increasing evidence that activity-dependent plasticity also involves the formation and disassembly of synapses. While we are gaining insight into the mechanisms of both synapse assembly and disassembly, we understand very little about how these phenomena are related to each other and how they might be coordinately controlled to achieve the precise patterns of synaptic connectivity in the nervous system. Here, we review our current understanding of both synapse assembly and disassembly in an effort to unravel the relationship between these fundamental developmental processes. PMID- 14556708 TI - The birth of a channel. AB - An ion channel protein begins life as a nascent peptide inside a ribosome, moves to the endoplasmic reticulum where it becomes integrated into the lipid bilayer, and ultimately forms a functional unit that conducts ions in a well-regulated fashion. Here, I discuss the nascent peptide and its tasks as it wends its way through ribosomal tunnels and exit ports, through translocons, and into the bilayer. We are just beginning to explore the sequence of these events, mechanisms of ion channel structure formation, when biogenic decisions are made, and by which participants. These decisions include when to exit the endoplasmic reticulum and with whom to associate. Such issues govern the expression of ion channels at the cell surface and thus the electrical activity of a cell. PMID- 14556709 TI - Neuronal polarity and trafficking. AB - Among the most morphologically complex cells, neurons are masters of membrane specialization. Nowhere is this more striking than in the division of cellular labor between the axon and the dendrites. In morphology, signaling properties, cytoskeletal organization, and physiological function, axons and dendrites (or more properly, the somatodendritic compartment) are radically different. Such polarization of neurons into domains specialized for either receiving (dendrites) or transmitting (axons) cellular signals provides the underpinning for all neural circuitry. The initial specification of axonal and dendritic identity occurs early in neuronal life, persists for decades, and is manifested by the presence of very different sets of cell surface proteins. Yet, how neuronal polarity is established, how distinct axonal and somatodendritic domains are maintained, and how integral membrane proteins are directed to dendrites or accumulate in axons remain enduring and formidable questions in neuronal cell biology. PMID- 14556710 TI - Polarized domains of myelinated axons. AB - The entire length of myelinated axons is organized into a series of polarized domains that center around nodes of Ranvier. These domains, which are crucial for normal saltatory conduction, consist of distinct multiprotein complexes of cell adhesion molecules, ion channels, and scaffolding molecules; they also differ in their diameter, organelle content, and rates of axonal transport. Juxtacrine signals from myelinating glia direct their sequential assembly. The composition, mechanisms of assembly, and function of these molecular domains will be reviewed. I also discuss similarities of this domain organization to that of polarized epithelia and present emerging evidence that disorders of domain organization and function contribute to the axonopathies of myelin and other neurologic disorders. PMID- 14556711 TI - Specific localization and timing in neuronal signal transduction mediated by protein-lipid interactions. AB - A large number of signaling proteins translocate from the cytosol to the plasma membrane in response to receptor and electrical stimuli. The site of translocation to the plasma membrane and the "on" and "off" rates of the translocation process are critical for defining the specificity of the signaling response. In addition to targeting mechanisms based on protein-protein interactions, signaling proteins have evolved a large repertoire of covalent lipid modifications and lipid binding protein modules that regulate reversible membrane association. The time constants of these membrane interactions range from milliseconds to several hours. Here we discuss how diversity in lipid-based membrane anchoring and targeting motifs contributes to plasticity in neuronal signaling by providing local and regional control mechanisms as well as a means to transduce and integrate signals over a broad range of different time scales. PMID- 14556712 TI - Local calcium signaling in neurons. AB - Transient rises in the cytoplasmic concentration of calcium ions serve as second messenger signals that control many neuronal functions. Selective triggering of these functions is achieved through spatial localization of calcium signals. Several qualitatively different forms of local calcium signaling can be distinguished by the location of open calcium channels as well as by the distance between these channels and the calcium binding proteins that serve as the molecular targets of calcium action. Local calcium signaling is especially prominent at presynaptic active zones and postsynaptic densities, structures that are distinguished by highly organized macromolecular arrays that yield precise spatial arrangements of calcium signaling proteins. Similar forms of local calcium signaling may be employed throughout the nervous system, though much remains to be learned about the molecular underpinnings of these events. PMID- 14556713 TI - Compartmentalized synthesis and degradation of proteins in neurons. AB - An important aspect of gene expression in neurons involves the delivery of mRNAs to particular subcellular domains, where translation of the mRNAs is locally controlled. Local synthesis of protein within dendrites plays a key role in activity-dependent synaptic modifications. In growing axons, local synthesis in the growth cone is important for extension and guidance. Recent evidence also documents the existence of mechanisms permitting local protein degradation, providing bidirectional control of protein composition in local domains. Here, we summarize what is known about local synthesis and degradation of protein in dendrites and axons, highlighting key unresolved questions. PMID- 14556714 TI - AMPA receptor trafficking at excitatory synapses. AB - Excitatory synapses in the CNS release glutamate, which acts primarily on two sides of ionotropic receptors: AMPA receptors and NMDA receptors. AMPA receptors mediate the postsynaptic depolarization that initiates neuronal firing, whereas NMDA receptors initiate synaptic plasticity. Recent studies have emphasized that distinct mechanisms control synaptic expression of these two receptor classes. Whereas NMDA receptor proteins are relatively fixed, AMPA receptors cycle synaptic membranes on and off. A large family of interacting proteins regulates AMPA receptor turnover at synapses and thereby influences synaptic strength. Furthermore, neuronal activity controls synaptic AMPA receptor trafficking, and this dynamic process plays a key role in the synaptic plasticity that is thought to underlie aspects of learning and memory. PMID- 14556715 TI - Neurotransmitter release at central synapses. AB - Our understanding of synaptic transmission has grown dramatically during the 15 years since the first issue of Neuron was published, a growth rate expected from the rapid progress in modern biology. As in all of biology, new techniques have led to major advances in the cell and molecular biology of synapses, and the subject has evolved in ways (like the production of genetically engineered mice) that could not even be imagined 15 years ago. My plan for this review is to summarize what we knew about neurotransmitter release when Neuron first appeared and what we recognized we did not know, and then to describe how our views have changed in the intervening decade and a half. Some things we knew about synapses- "knew" in the sense that the field had reached a consensus--are no longer accepted, but for the most part, impressive advances have led to a new consensus on many issues. What I find fascinating is that in certain ways nothing has changed--many of the old arguments persist or recur in a different guise--but in other ways the field would be unrecognizable to a neurobiologist time-transported from 1988 to 2003. PMID- 14556716 TI - Glial cells and neurotransmission: an inclusive view of synaptic function. AB - Glial cells throughout the nervous system are closely associated with synapses. Accompanying these anatomical couplings are intriguing functional interactions, including the capacity of certain glial cells to respond to and modulate neurotransmission. Glial cells can also help establish, maintain, and reconstitute synapses. In this review, we discuss evidence indicating that glial cells make important contributions to synaptic function. PMID- 14556717 TI - Diversity in the mechanisms of neuronal cell death. AB - Neurons may die as a normal physiological process during development or as a pathological process in diseases. The best-understood mechanism of neuronal cell death is apoptosis, which is regulated by an evolutionarily conserved cellular pathway that consists of the caspase family, the Bcl-2 family, and the adaptor protein Apaf-1. Apoptosis, however, may not be the only cellular mechanism that regulates neuronal cell death. Neuronal cell death may exhibit morphological features of autophagy or necrosis, which differ from that of the canonical apoptosis. This review evaluates the evidence supporting the existence of alternative mechanisms of neuronal cell death and proposes the possible existence of an evolutionarily conserved pathway of necrosis. PMID- 14556718 TI - Do disorders of movement cause movement disorders and dementia? AB - Neurons require long-distance microtubule-based transport systems to ferry vital cellular cargoes and signals between cell bodies and axonal or dendritic terminals. Considerable progress has been made on developing a molecular understanding of these processes and how they are integrated into normal neuronal functions. Recent work also suggests that these transport systems may fail early in the pathogenesis of a number of neurodegenerative diseases. PMID- 14556719 TI - The ubiquitin proteasome system in neurodegenerative diseases: sometimes the chicken, sometimes the egg. AB - The ubiquitin-proteasome system targets numerous cellular proteins for degradation. In addition, modifications by ubiquitin-like proteins as well as proteins containing ubiquitin-interacting and -associated motifs modulate many others. This tightly controlled process involves multiple specific and general enzymes of the system as well as many modifying and ancillary proteins. Thus, it is not surprising that ubiquitin-mediated degradation/processing/modification regulates a broad array of basic cellular processes. Moreover, aberrations in the system have been implicated, either as a primary cause or secondary consequence, in the pathogenesis of both inherited and acquired neurodegenerative diseases. Recent findings indicate that the system is involved in the pathogenesis of Parkinson's, Alzheimer's, Huntington's, and Prion diseases as well as amyotrophic lateral sclerosis. This raises hopes for a better understanding of the pathogenetic mechanisms involved in these diseases and for the development of novel, mechanism-based therapeutic modalities. PMID- 14556720 TI - Nitric oxide, inflammation and acute burn injury. AB - Nitric oxide (NOz.rad;) is a diatomic mediator liberated on oxidation of L arginine by the nitric oxide synthase (NOS) family of enzymes. It has complex and wide ranging functions in vivo and has been implicated in the development of the profound inflammatory response that occurs as a result of cutaneous burn injury. In addition, dysregulation of NOS activity has been associated with multiple organ failure in human burn patients and may therefore represent a novel therapeutic target in such circumstances. This review focuses on the role of NOz.rad; in inflammation, with particular emphasis on the acute post-burn inflammatory response. Specific areas of discussion include the maintenance of microvascular haemostasis, leukocyte recruitment and remote organ dysfunction following thermal injury. PMID- 14556721 TI - Monocyte chemoattractant protein-1 (MCP-1) and macrophage infiltration into the skin after burn injury in aged mice. AB - Clinical observations and laboratory studies have shown a delay in dermal wound healing in aged subjects. Since macrophages play a key role in wound healing, we investigated age related differences in MCP-1 production and monocyte recruitment to the wound following burn injury using a murine model. The present study shows that there is an increase in MCP-1 levels in the burned-normal skin interface at 1-day post burn in both young and aged burned mice compared to sham injured mice. However, the levels of MCP-1 in aged burned mice (133.16+/-36.55pg/mg protein) were approximately half the levels of young burned mice (286.15+/-45.36pg/mg protein, P<0.05). Additionally, at 4 days post burn, MCP-1 levels in aged mice (290.73+/-101.98) reached the same levels as in young mice (243.97+/-36.71). There was no difference in macrophage accumulation into the wound between young and aged at either time point. These data demonstrate that the difference in dermal MCP-1 levels between the young and aged is not associated with a difference in macrophage infiltration to the wound following burn injury, suggesting that the lower MCP-1 content in the aged is possibly affecting other phases of wound healing in the aged. PMID- 14556722 TI - The female, red Duroc pig as an animal model of hypertrophic scarring and the potential role of the cones of skin. AB - Hypertrophic scarring occurs after deep dermal wounds. Our understanding of the etiology is poor; one reason is the lack of an animal model. In 1972, Silverstein described scarring in the Duroc pig but the model was never confirmed nor disproved. Another reason, as we previously suggested, is that hypertrophic scarring only occurs within regions of human skin that contain cones and the cones have not been studied in relation to hypertrophic scarring. We, therefore (i) explored healing in the female, red Duroc model for similarities to human hypertrophic scarring, studying wound thickness, appearance, healing status at 3 weeks, histology, and immunocytochemical localization of decorin, versican, TGFbeta1 and IGF-1; and (ii) examined Duroc skin for cones. We found that healing after deep wounds in Duroc pigs is similar, but not identical, to human hypertrophic scarring. We also found that Duroc skin contains cones. Healing in the female, red Duroc pig is sufficiently similar to human hypertrophic scarring to warrant further study so that it can be accepted or rejected as a model of human hypertrophic scarring. In addition, the relationship of the cones to hypertrophic scarring needs further detail and can be studied in this model. PMID- 14556723 TI - Laser Doppler imaging determines need for excision and grafting in advance of clinical judgment: a prospective blinded trial. AB - INTRODUCTION: Clinicians' judgment as to which burns require excision and grafting remains one aspect of burn care without objective measurements. This study presents a prospective, blinded trial to assess decision to operate by laser Doppler imaging (numerical criteria) versus the clinical judgment of an experienced burn surgeon. METHODS: A number of 23 patients were enrolled in this prospective trial and 41 representative wounds of indeterminate depth were selected for observation. Daily determination of need to operate (burn depth) was made by a single burn surgeon. Laser Doppler imager (LDI) scans of the same wounds were simultaneously obtained, and not revealed to the clinician. Data analysis compared quickness of decision to operate by LDI to the clinician's judgment. Concurrence of decisions by either method was compared. RESULTS: A total of 23 patients and 41 wounds were analyzed. LDI and the surgeon agreed in determination of wound depth 56% of the time (23/41, P=0.031). Biopsy confirmation was obtained for 21 wounds. The surgeon's determination of burn depth was accurate in 71.4% of wounds biopsied (15/21). When the LDI scan median flux indicated need for excision, it was 100% accurate (7/7). When both the surgeon and the LDI were correct in assessing wound depth, LDI would have saved median number of 2 days (minimum=0, maximum=4). CONCLUSION: LDI allowed for earlier, objective determination of need to operate. Concurrence with clinical judgment in this blinded study was excellent. LDI should be seen as an effective aid to clinical judgment when contemplating excision of burns with indeterminate depth. PMID- 14556724 TI - Impairment after burns: a two-center, prospective report. AB - Impairment rating is regularly reported for trauma and other conditions but rarely for burns. The purposes of this study were: (1) to report impairment collected prospectively at our burn center, (2) to relate this impairment to measures of psychosocial and functional outcome, and (3) to compare these data to similar data from another burn center to verify that rating impairment is standardized and that the impairments are similar. We studied 139 patients from the University of Washington (UW) Burn Center and 100 patients from the University of Texas (UT) Southwestern Burn Center. The average whole person impairment (WPI) ratings at the University of Washington were 17% and this correlated with total body surface area burned and days off work. It did not correlate with Brief Symptom Inventory (BSI), Functional Independence Measure (FIM), Short-Form 36-Item Health Survey (SF-36), Satisfaction With Life Scale (SWLS), and the Community Integration Questionnaire (CIQ). Average whole person impairment ratings at UT Southwestern were similar at 19%. Several components of the impairment rating, however, differed at the two institutions. To minimize this variation, we recommend: (1) use the skin impairment definitions of the fifth edition of the Guides to the Evaluation of Permanent Impairment (or the most recent published versions of the Guide), and (2) include sensory impairment in healed burns and skin grafts in the skin impairment. PMID- 14556725 TI - A comparison of keratinocyte cell sprays with and without fibrin glue. AB - Fibrin glue is an excellent template for cellular migration and has been shown to be an effective delivery system for cultured autologous keratinocytes. We have investigated whether fibrin glue has any benefit on the percentage of epithelial cover when cultured autologous keratinocytes are sprayed onto a freshly debrided wound bed. Three pigs were used for this study. This provided a total of 18 full thickness, vertically orientated wounds, each 4cm in diameter and isolated in PTFE chambers to prevent re-epithelialisation from the wound margins. Eight wounds were sprayed with cultured autologous keratinocytes suspended in 2ml culture medium and eight wounds were sprayed with cultured autologous keratinocytes suspended in 1ml of the fibrin/aprotinin component of Tisseel fibrin glue (Baxter) mixed with 1ml of culture medium. In the latter group the thrombin component of the fibrin glue kit was applied to the wound bed immediately prior to grafting. The remaining two wounds were used as controls and sprayed with either culture medium or fibrin glue without cells. Epithelial cover was calculated in whole-wound biopsies at 3 weeks using image analysis, histology and immunohistochemistry. The cell suspension in fibrin glue appeared to spread more evenly over the wound surface, with no pooling in the inferior aspect of the wound. However, mean epithelial area at 3 weeks in the fibrin group was 1.6cm(2) per wound compared with 1.8cm(2) for the non-fibrin group, as measured by image analysis of digital photographs. There was no statistically significant difference between the two groups (P=0.802). This surprising result was confirmed by histological analysis of the wound biopsies, with a good correlation between histological and image analysis data (R=0.967). There was no observable difference in the quality of the epithelium on histological and immunohistological analysis of either group. PMID- 14556726 TI - An epidemiologic study of elderly burn patients in Ain Shams University Burn Unit, Cairo, Egypt. AB - Recent statistics from Egypt indicate that elderly persons comprise 5.8% of the general population. Elderly patients remain a high-risk group in all burn units. This prospective study investigated geriatric burn patients (defined as 60 years and older) who were present at our center between May 1995 and October 2001. Of the 4220 patients who were present during this period, 97 (2.3%) were elderly, and 63 (7.1%) of the 880 total admitted were geriatrics. Of the 97 elderly burn patients who were present during the study period, 31 died (mortality rate 31.9%). Other epidemiological data, including duration of hospitalization, seasonal and day/hour variation in burn incidence and burn causes and accident site are presented. Ideas for management and prevention are also presented. PMID- 14556727 TI - Characteristics of self-immolation attempts in Akita Prefecture, Japan. AB - Akita Prefecture has had the highest suicide rate in Japan for the past 9 years. To obtain further information on suicide attempts by self-immolation in Akita, we performed a statistical analysis of patients in this prefecture who attempted to burn themselves. Over the past 6 years, 541 patients suffering from burns were transferred to medical emergency units. Of these, 35 (6.5%) attempted self immolation, most of whom were between 20 and 60 years of age. Women over 50 years of age outnumbered men in the same age group. All 35 patients sustained flame burns. The total burn surface area (TBSA), burn index (BI), rate of inhalation injury, and mortality rate were all significantly higher in the patients who attempted self-immolation than in those with nonsuicidal burns. Most (68.6%) of the self-immolation attempts were made indoors. Because the Japanese are not generally a very religious people, training to help them cultivate a philosophy of life and educating them in moral science to help them form a personal view of life and death may be necessary to prevent suicides. PMID- 14556728 TI - Comparison of self-reported and monitored compliance of daily injection of human growth hormone in burned children. AB - Objectives. We compared insulin-like growth factor (IGF-I) levels obtained in two groups with different methods of assessing compliance. Burned children were randomized to receive a daily injection of 0.05-0.1mg/kg per day of recombinant human growth hormone (rhGH) or placebo. Study design. One hundred twenty-five children (age range 6 months to 17 years) with total body surface area burns >40% participated in the study. Baseline levels of IGF-I were obtained at hospital discharge, at which time daily injections of rhGH or placebo were initiated. Assessment of IGF-I levels was repeated at 3-month intervals for 1 year. A directly monitored group met daily with research staff that witnessed the preparation and injection of the study drug. A self-reported group completed a Self Reported Compliance Questionnaire (SRCQ) that assessed compliance with drug regimen. A compliance of at least 85% (injections reported administered/maximal total injections possible) was accepted as being compliant. Data were analyzed using a one-way ANOVA followed by a Student Newman-Kuels test, with the results given as means+/-S.E.M. Results. The percent change in IGF-I levels between 6 and 9 months in the self-reported (22.7+/-7.9%) and monitored groups (21.9+/-8.6%) were similar. In contrast, the percent change in IGF-I levels in the placebo group was significantly decreased (-5.6+/-6.3%). Conclusions. Self-reported scores via an SRCQ is a potentially useful and valid method of assessing compliance of rhGH injections, as both reported and directly monitored methods yield similar changes in levels of IGF-I. PMID- 14556729 TI - A randomised clinical trial comparing a hydrocolloid-derived dressing and glycerol preserved allograft skin in the management of partial thickness burns. AB - Membranous dressings for the treatment of partial and mixed thickness burns are among the most innovative and promising new developments of the last years. In this study, we present data of a randomised prospective comparative study on a carboxymethylcellulose based dressing, Hydrofibre((R)) and glycerolized human allograft skin. In a 2 year period, 80 patients (40 for each material) were enrolled in the trial. Study wounds (<10% TBSA) that had not re-epithelialised after 14+/-3 days were debrided and grafted or, if small enough, managed with a topical antimicrobial agent. Mean total TBSA was 8.3+/-5.2%, study burn 3.7+/ 2.0% for the Hydrofibre((R)) group and 7.3+/-4.3% total, 3.4+/-2.1% study burn for the allograft skin group (n.s. Wilcoxon rank sum test). No significant differences between groups were established in number of patients with superficial/deep burns. In both groups about 2/3 of the patients healed completely with the dressings applied (24/40 versus 27/40 for Hydrofibre((R)) versus allograft skin, respectively). However, a higher incidence of post-study excision and grafting was found in the Hydrofibre((R)) group (45% versus 15% in the allograft skin group, P=0.004, Mann-Whitney). At 10 weeks follow-up no significant differences were seen in scar colour, pigmentation, pliability, height or itching (Vancouver Scar Scale). Skin elasticity, measured by the Cutometer((R)), was significantly better for the allograft group (P=0.010, Wilcoxon). These differences were no longer found at 6 months and 1 year follow up. Incidence of hypertrophy after 6 months was higher, but not significantly, in the Hydrofibre((R)) compared to the allograft skin group (52.5% versus 30%, P=0.09, chi-square). In view of the results from our comparative study on Hydrofibre((R)) versus allograft skin, we prefer the use of allograft skin for the category of larger burns of mixed depth, usually presented to burn centres. However, for partial thickness and small burns Hydrofibre((R)) can be the first choice in treatment. PMID- 14556730 TI - Stenotrophomonas maltophilia: a serious and rare complication in patients suffering from burns. AB - Stenotrophomonas maltophilia is rarely implicated in clinical infections but it constitutes a significant nosocomial pathogen, especially in immunocompromised patients. This report describes the first case of a generalised infection caused by S. maltophilia that included bacteremia, wound and respiratory tract infection in a patient suffering from burns. Given the emergence of S. maltophilia nosocomial infections, especially in patients with burns, isolation of the bacterium from blood cultures should prompt the commencement of adequate antibiotic treatment. PMID- 14556731 TI - Paediatric friction hand injuries caused by domestic vacuum cleaners-a review from one unit. AB - The management of seven children with vacuum cleaner friction burns is described. Six of the children sustained full thickness burns. Four were treated operatively. Indications for surgery are outlined and important accident prevention strategies are highlighted. PMID- 14556733 TI - Double reverse V-Y-plasty in postburn scar contractures: a new modification of V Y-plasty. AB - Several techniques have been defined to use in surgical treatment of postburn scar contractures. However, distal flap necrosis is frequently seen since most of these techniques require random-pattern flaps and there is often poor vascular supply to scar tissue. In the Z-plasty, V-Y-plasty and their analogues, excess tissue requires excision of the dog-ear. A new modification of V-Y-plasty, called "double reverse V-Y-plasty", is discussed. Nineteen postburn scar contractures were successfully treated with double reverse V-Y-plasty. The postoperative results represent the versatility of this technique in the surgical treatment of postburn scar contractures, especially neck and extremities. There was no distal flap necrosis. Double reverse V-Y-plasty is effective and alternative to the current techniques in surgical treatment of every kind of postburn scar contractures with one or more contracture lines. It does not need any surgical knack. ADVANTAGES: (i) double reverse V-Y-plasty is safely useful, when skin tension across the contracture line is too great to use any local flaps; (ii) when superficial scarring is localized in the contracture site, it is superior to other local flaps because of rich vascularity and mobility; (iii) double reverse V-Y-plasty is advised to the inexperienced surgeon, since it is easy to use; (iv) color and texture matches are cosmetically acceptable, and the resultant contracture is as much as with other techniques; (v) there is no necessity to use excision of the dog-ear; (vi) it can be utilized under local anesthesia almost in all cases; and (vii) it requires a shorter period of operation, and hospitalization. PMID- 14556732 TI - Hydrocortisone improved haemodynamics and fluid requirement in surviving but not non-surviving of severely burned patients. AB - Recent studies have shown that administration of hydrocortisone may lead to a reduction of catecholamines and to an improved outcome in septic patients. However, there are no data on the use of hydrocortisone in burn patients although in these patients reduction of vasopressors might be even more crucial for outcome due to improvement of skin perfusion. This study presents the first results on the impact of hydrocortisone administration in norepinephrine dependent severely burned patients. In a prospective cohort study fourteen consecutive severely burned patients received, 12h after norepinephrine dependency, a hydrocortisone bolus of 100mg followed by 0.18mg/(kgh) hydrocortisone. The course of the necessary norepinephrine dose, as well as the fluid balance was documented 12h prior and after the first dosage of hydrocortisone. Statistical analysis showed an unexpected increase of the required norepinephrine dosage. A statistical post hoc evaluation of surviving and non-surviving patients revealed a significant increase of norepinephrine in non-survivors whereas in survivors it was possible to reduce norepinephrine significantly. Furthermore, the median fluid requirement of surviving patients could be significantly reduced whereas in the group of non-survivors there was no change of volume needed. Our data suggests that hydrocortisone might be useful in selected patients with severe burn injuries. However, patients not responding to hydrocortisone administrations seem to have a poor prognosis. Our findings are in contrast to previously published data on septic patients, in whom hydrocortisone administration resulted in a reduction of norepinephrine. In burned patients the severity of trauma seems to have more profound influence on the pathophysiological mechanism of sepsis. Due to the high number of non-responders, the potential immune suppression and impaired wound healing caused by the side effects of hydrocortisone, further selection criteria seem to be necessary. A short ACTH-test might be considered prior to hydrocortisone administration to select patients who might benefit from this therapy. In summary, further prospective controlled studies will be necessary to establish hydrocortisone in the routine therapy of severely burned patients. PMID- 14556734 TI - Repair and reconstruction of massively damaged burn wounds. AB - OBJECTIVE: To report repair and reconstruction of massively damaged burn wounds under unusual condition. METHODS: One hundred forty-eight patients with deep burn tissue defects admitted from January 1993 through December 2000 were analyzed, among them 96 patients suffered from electrical injury which constituted 65.3% of all cases, 18 patients with hot press injury, 18 cases with deep burns as a result of CO poisoning or epileptic seizure, accounting for 12.2 and 12.2%, respectively, 6 cases with radiation injury, comprising 4.0% of all cases, 2 cases with explosive injury, 2 cases with chemical burn and 6 cases caused by erosive chemicals and wound infection. One hundred seventy-six flaps were transferred with mostly local flaps to repair deep burn wounds in 148 patients with tissue defects, in which necrotic tendons were replaced by acellular allogeneic tendons simultaneously in seven cases. Sixty-one iliolumbar arterial axial skin flaps for coverage of soft-tissue defects in hands or wrists were transplanted. Technical innovations to repair large soft-tissue defects of temporal region and ear, chin and lip, and dorso-lateral aspect of foot due to deep burn were explored. New technics to define necrotic tissue and vascular damage as a result of electrical injury were developed. RESULTS: The biggest dimension of flaps in this group was 22cmx30cm. The survival rate of flap was 96.5%, while necrosis of the tip of flap occurred in 3.5%. The function and configuration were satisfactory after 4 months to 8 years follow-up. The technique of 99Tcm-methylene di-phosphonate (99Tcm-MDP) scintigraphy helped identify necrotic tissue before operation, and with the help of digital subtraction arteriogram (DSA) arterial injury could be identified. B-mode ultrasound was helpful to show the extent of endothelial injury, and Colour Doppler was useful to show luminal blood flow signal and filling defect in the injured artery. CONCLUSIONS: Repair and reconstruction of massively damaged burn wound at early stage could be achieved. Techniques which helped define the extent of damage to the soft tissues, including arteries and tendons were essential preoperatively for successful reconstruction. Functional and aesthetic reconstruction, as well as the general condition of the patients, could thus be significantly improved early after the devastating injuries. Further innovations of operative technics would benefit more patients with such injuries. PMID- 14556735 TI - Bacterial toxicosis/toxic shock syndrome as a contributor to morbidity in children with burn injuries. AB - When a child with a thermal injury suddenly becomes unwell, it is often difficult to obtain an accurate diagnosis in the early stages of the illness. Out of 71 children admitted to our burns unit over a 15-month period, 13 became acutely toxic. Most of the cases had relatively small burns and the exact reason for their sudden deterioration was not apparent in the immediate stages. Several children required admission to the intensive therapy unit and there was one death. The number of children experiencing this significant morbidity over a relatively short period of time prompted us to perform a detailed retrospective study to ascertain the exact cause of the deterioration and to identify any patterns in their presenting signs and symptoms. There was considerable variation in the clinical picture of the children who developed the toxicosis, however, several features were frequently observed which resembled those seen in the toxic shock syndrome (TSS). In order to be more specific, we applied the criteria for TSS as defined by the Centres for Disease Control (CDC) to each of the 13 children. All of the criteria were noted in six children and the majority in the other seven. This was a much higher incidence than we would normally have expected. The clinical features of children with TSS are not always easy to distinguish from those observed in other illnesses. We therefore explored the possibility of alternative diagnoses that may have caused the toxicosis in these children. The mortality associated with TSS can be high, especially if there is a delay in recognition and subsequent management of the disease. Clinicians should be alerted to the fact that TSS is probably more prevalent than previously thought and should practise with a high index of suspicion in any child with a burn whose clinical condition suddenly deteriorates. PMID- 14556736 TI - Frostbite of the gluteal region. PMID- 14556737 TI - The use of an island posterior calf fasciocutaneous flap for sensate cover of the burnt knee. PMID- 14556738 TI - Mud volcano and deep dermal burns. PMID- 14556739 TI - Sites for interaction between Gal80p and Gal1p in Kluyveromyces lactis: structural model of galactokinase based on homology to the GHMP protein family. AB - The induction of transcription of the galactose genes in yeast involves the galactose-dependent binding of ScGal3p (in Saccharomyces cerevisiae) or KlGal1p (in Kluyveromyces lactis) to Gal80p. This binding abrogates Gal80's inhibitory effect on the activation domain of Gal4p, which can then activate transcription. Here, we describe the isolation and characterization of new interaction mutants of K.lactis GAL1 and GAL80 using a two-hybrid screen. We present the first structural model for Gal1p to be based on the published crystal structures of other proteins belonging to the GHMP (galactokinase, homoserine kinase, mevalonate kinase and phosphomevalonate kinase) kinase family and our own X-ray diffraction data of Gal1p crystals at 3A resolution. The locations of the various mutations in the modelled Gal1p structure identify domains involved in the interaction with Gal80p and provide a structural explanation for the phenotype of constitutive GAL1 mutations. PMID- 14556740 TI - Molecular dynamics simulation of dark-adapted rhodopsin in an explicit membrane bilayer: coupling between local retinal and larger scale conformational change. AB - The light-driven photocycle of rhodopsin begins the photoreceptor cascade that underlies visual response. In a sequence of events, the retinal covalently attached to the rhodopsin protein undergoes a conformational change that communicates local changes to a global conformational change throughout the whole protein. In turn, the large-scale protein change then activates G-proteins and signal amplification throughout the cell. The nature of this change, involving a coupling between a local process and larger changes throughout the protein, may be important for many membrane proteins. In addition, functional work has shown that this coupling occurs with different efficiency in different lipid settings. To begin to understand the nature of the efficiency of this coupling in different lipid settings, we present a molecular dynamics study of rhodopsin in an explicit dioleoyl-phosphatidylcholine bilayer. Our system was simulated for 40 ns and provides insights into the very early events of the visual cascade, before the full transition and activation have occurred. In particular, we see an event near 10 ns that begins with a change in hydrogen bonding near the retinal and that leads through a series of coupled changes to a shift in helical tilt. This type of event, though rare on the molecular dynamics time-scale, could be an important clue to the types of coupling that occur between local and large-scale conformational change in many membrane proteins. PMID- 14556741 TI - Crystal structure of a family 4 uracil-DNA glycosylase from Thermus thermophilus HB8. AB - Uracil-DNA glycosylase (UDG; EC 3.2.2.-) removes uracil from DNA to initiate DNA base excision repair. Since hydrolytic deamination of cytosine to uracil is one of the most frequent DNA-damaging events in all cells, UDG is an essential enzyme for maintaining the integrity of genomic information. For the first time, we report the crystal structure of a family 4 UDG from Thermus thermophilus HB8 (TthUDG) complexed with uracil, solved at 1.5 angstroms resolution. As opposed to UDG enzymes in its other families, TthUDG possesses a [4Fe-4S] cluster. This iron sulfur cluster, which is distant from the active site, interacts with loop structures and has been suggested to be unessential to the activity but necessary for stabilizing the loop structures. In addition to the iron-sulfur cluster, salt bridges and ion pairs on the molecular surface and the presence of proline on loops and turns is thought to contribute to the enzyme's thermostability. Despite very low levels of sequence identity with Escherichia coli and human UDGs (family 1) and E.coli G:T/U mismatch-specific DNA glycosylase (MUG) (family 2), the topology and order of secondary structures of TthUDG are similar to those of these distant relatives. Furthermore, the coordinates of the core structure formed by beta-strands are almost the same. Positive charge is distributed over the active-site groove, where TthUDG would bind DNA strands, as do UDG enzymes in other families. TthUDG recognizes uracil specifically in the same manner as does human UDG (family 1), rather than guanine in the complementary strand DNA, as does E.coli MUG (family 2). These results suggest that the mechanism by which family 4 UDGs remove uracils from DNA is similar to that of family 1 enzymes. PMID- 14556742 TI - Mechanism of C2-toxin inhibition by fluphenazine and related compounds: investigation of their binding kinetics to the C2II-channel using the current noise analysis. AB - The binding component C2II of the binary actin ADP-ribosylating C2-toxin from Clostridium botulinum is essential for intoxication of target cells. Activation by a protease leads to channel formation and this is presumably required for the transport of the toxic C2I component into cells. The C2II-channel is cation selective and contains a binding site for fluphenazine and structurally related compounds. Ion transport through C2II and in vivo intoxication is blocked when the sites are occupied by the ligands. C2II was reconstituted into artificial lipid bilayer membranes and formed ion permeable channels. The binding constant of chloroquine, primaquine, quinacrine, chloropromazine and fluphenazine to the C2II-channel was determined using titration experiments, which resulted in its block. The ligand-induced current noise of the C2II-channels was investigated using fast Fourier transformation. The noise of the open channels had a rather small spectral density, which was a function of the inverse frequency up to about 100 Hz. Upon addition of ligands to the aqueous phase the current through C2II decreased in a dose-dependent manner. Simultaneously, the spectral density of the current noise increased drastically and its frequency dependence was of Lorentzian type, which was caused by the on and off-reactions of the ligand mediated channel block. The ligand-induced current noise of C2II was used for the evaluation of the binding kinetics for different ligands to the channel. The on rate constant of ligand binding was between 10(7) and 10(9) M(-1) s(-1) and was dependent on the ionic strength of the aqueous phase. The off-rate varied between about 10 s(-1) and 3900 s(-1) and depended on the structure of the ligand. The role of structural requirements for the effective block of C2II by the different ligands is discussed. PMID- 14556743 TI - A structural analysis of the interaction between ncd tail and tubulin protofilaments. AB - ncd is a minus-end directed, kinesin-like motor, which binds to microtubules with its motor domain and its cargo domain as well. Typical of retrograde motors, the motor domain of ncd locates to the C-terminal end of the polypeptide chain, and hence, the cargo domain constitutes the N-terminal region. To date, several studies have investigated the interaction properties of the motor domain with microtubules, but very few structural data are available about the tail itself or its interaction with microtubules as cargo. Here, we applied cryo-electron microscopy and helical 3D image reconstruction to 15 protofilament microtubules decorated with an ncd tail fragment (N-terminal residues 83-187, named NT6). In our study, the ncd tail shows a behaviour resembling filamentous MAPs such as tau protein, exhibiting a highly flexible structure with no large globular domains. NT6 binds to four different sites on the outer side of microtubules within the proximity of the kinesin motor-binding site. Two of these sites locate within the groove between two neighbouring protofilaments, and appear as strong binding sites, while the other two sites, located at the outer rim, appear to play a secondary role. In addition, the ncd tail fragment induces the formation of large protofilament sheets, suggesting a tail-induced modification of lateral protofilament contacts. PMID- 14556744 TI - Synchronous loss of quasispecies memory in parallel viral lineages: a deterministic feature of viral quasispecies. AB - Viral quasispecies are endowed with a memory of their past evolutionary history in the form of minority genomes of their mutant spectra. To determine the fate of memory genomes in evolving viral quasispecies, we have measured memory levels of antigenic variant of foot-and-mouth disease virus (FMDV) RED, which includes an Arg-Glu-Asp (RED) at a surface antigenic loop of the viral capsid. The RED reverted to the standard Arg-Gly-Asp (RGD), and the RED remained as memory in the evolving quasispecies. In four parallel evolutionary lineages, memory reduction followed a strikingly similar pattern, and at passage 60 memory levels were indistinguishable from those of control populations (devoid of memory). Nucleotide sequence analyses indicated that memory loss occurred synchronously despite its ultimate molecular basis being the stochastic occurrence of mutations in the evolving quasispecies. These results on the kinetics of memory levels have unveiled a deterministic feature of viral quasispecies. Molecular mechanisms that may underlie synchronous memory loss are the averaging of noise signals derived from mutational input, and constraints to genome diversification imposed by a nucleotide sequence context in the viral genome. Possible implications of the behaviour of complex, adaptive viral systems as experimental models to address primary mechanisms of neurological memory are discussed. PMID- 14556745 TI - Linear remodeling of helical virus by movement protein binding. AB - Previously we have shown that encapsidated potato virus X (PVX) RNA was nontranslatable in vitro, but could be converted into a translatable form by binding of the PVX-coded movement protein (termed TGBp1) to one end of a polar helical PVX virion. We reported that binding of TGBp1 to coat protein (CP) subunits located at one extremity of the helical particles induced a linear destabilization of the CP helix, which was transmitted along the whole particle. Two model structures were used: (i) native PVX and (ii) artificial polar helical PVX-like particles lacking intact RNA (PVX(RNA-DEG)). Binding of TGBp1 to the end of either of these particles led to their destabilization, but no disassembly of the CP helix occurred. Influence of additional factors was required to trigger rapid disassembly of TGBp1-PVX and TGBp1-PVX(RNA-DEG) complexes. Thus: (i) no disassembly was observed unless TGBp1-PVX complex was translated. A novel phenomenon of TGBp1-dependent, ribosome-triggered disassembly of PVX was described: initiation of translation and few translocation steps were needed to trigger rapid (and presumably cooperative) disassembly of TGBp1-PVX into protein subunits and RNA. Importantly, the whole of the RNA molecule (including its 3' terminal region) was released. The TGBp1-induced linear destabilization of CP helix was reversible, suggesting that PVX in TGBp1-PVX complex was metastable; (ii) entire disassembly of the TGBp1-PVX(RNA-DEG) complex (but not of the TGBp1 free PVX(RNA-DEG) particles) into 2.8S subunits was triggered under influence of a centrifugal field. To our knowledge, transmission of the linear destabilization along the polar helical protein array induced by a foreign protein binding to the end of the helix represents a novel phenomenon. It is tempting to suggest that binding of TGBp1 to the end of the PVX CP helix induced conformational changes in terminal CP subunits that can be linearly transferred along the whole helical particle, i.e. that intersubunit conformational changes may be transferred along the CP helix. PMID- 14556746 TI - Tertiary structure of thiopurine methyltransferase from Pseudomonas syringae, a bacterial orthologue of a polymorphic, drug-metabolizing enzyme. AB - In humans, the enzyme thiopurine methyltransferase (TPMT) metabolizes 6 thiopurine (6-TP) medications, including 6-thioguanine, 6-mercaptopurine and azathioprine, commonly used for immune suppression and for the treatment of hematopoietic malignancies. S-Methylation by TPMT prevents the intracellular conversion of these drugs into active 6-thioguanine nucleotides (6-TGNs). Genetic polymorphisms in the TPMT protein sequence have been associated with decreased tissue enzymatic activities and an increased risk of life-threatening myelo suppression from standard doses of 6-TP medications. Biochemical studies have demonstrated that TPMT deficiency is primarily associated with increased degradation of the polymorphic proteins through an ubiquitylation and proteasomal dependent pathway. We have now determined the tertiary structure of the bacterial orthologue of TPMT from Pseudomonas syringae using NMR spectroscopy. Bacterial TPMT similarly catalyzes the S-adenosylmethionine (SAM)-dependent transmethylation of 6-TPs and shares 45% similarity (33% identity) with the human enzyme. Initial studies revealed an unstructured N terminus, which was removed for structural studies and subsequently determined to be required for enzymatic activity. Despite lacking sequence similarity to any protein of known three dimensional structure, the tertiary structure of bacterial TPMT reveals a classical SAM-dependent methyltransferase topology, consisting of a seven stranded beta-sheet flanked by alpha-helices on both sides. However, some deviations from the consensus topology, along with multiple insertions of structural elements, are evident. A review of the many experimentally determined tertiary structures of SAM-dependent methyltransferases demonstrates that such structural deviations from the consensus topology are common and often functionally important. PMID- 14556747 TI - Solution NMR study of the interaction between NTF2 and nucleoporin FxFG repeats. AB - Interactions with nucleoporins containing FxFG repeat cores are crucial for the nuclear import of RanGDP mediated by nuclear transport factor 2 (NTF2). We describe here a solution NMR-based study that identifies primary and secondary FxFG-binding sites on NTF2 and accounts for a range of observations on the rate of NTF2 nuclear trafficking. We used three complementary NMR methods, namely amide group chemical shift titrations, NOE and cross-saturation measurements, to show that the major FxFG-binding site on the dimeric rat NTF2 (rNTF2) molecule is centred on Trp7 and is formed by residues from both NTF2 chains. A secondary FxFG binding site is located at the rNTF2 hydrophobic cavity and these two sites, together with a surface hydrophobic cluster centred on Trp112, merge into an elongated hydrophobic stripe on the rNTF2 surface. The primary site centred on Trp7 is lost in the rNTF2-W7A mutant that has been shown to bind FxFG nucleoporins with greatly reduced affinity, whereas the secondary site at the rNTF2 hydrophobic cavity is retained. The interface between NTF2 and FxFG nucleoporins detected by NMR is more extensive than that detected by X-ray crystallography, and the presence of a secondary site at the NTF2 hydrophobic cavity accounts for the unexpectedly rapid nuclear import of rNTF2-W7R recently observed by others. The structure of the binding interfaces on these transport factors provides a rationale for the specificity of their interactions with nucleoporins that, combined with their weak binding constants, facilitates rapid translocation through NPCs during nuclear trafficking. PMID- 14556748 TI - Structural stability and solution structure of chaperonin GroES heptamer studied by synchrotron small-angle X-ray scattering. AB - The GroES protein from Escherichia coli is a well-known member of the molecular chaperones. GroES consists of seven identical 10 kDa subunits, and forms a dome like oligomeric structure. In order to obtain information on the structural stability and unfolding-refolding mechanism of GroES protein, especially at protein concentrations (0.4-1.2 mM GroES monomer) that would mimic heat stress conditions in vivo, we have performed synchrotron small-angle X-ray scattering (SAXS) experiments. Surprisingly, in spite of the high protein concentration, reversibility in the unfolding-refolding reaction was confirmed by SAXS experiments structurally. Although the unfolding-refolding reaction showed an apparent single transition with a Cm of 1.1 M guanidium hydrochloride, a more detailed analysis of this transition demonstrated that the unfolding mechanism could be best explained by a sequential three-state model, which consists of native heptamer, dissociated monomer, and unfolded monomer. Together with our previous result that GroES unfolded completely via a partially folded monomer according to a three-state model at low protein concentration (5 microM monomer), the unfolding-refolding mechanism of GroES protein could be explained uniformly by the three-state model from low to high protein concentrations. Furthermore, to clarify an ambiguity of the native GroES structure in solution, especially mobile loop structures, we have estimated a solution structure of GroES using SAXS profiles obtained from experiments and simulation analysis. The result suggested that the native structure of GroES in solution was very similar to that seen in GroES-GroEL complex determined by crystallography. PMID- 14556749 TI - Functional recycling of C2 domains throughout evolution: a comparative study of synaptotagmin, protein kinase C and phospholipase C by sequence, structural and modelling approaches. AB - The C2 domain is one of the most frequent and widely distributed calcium-binding motifs. Its structure comprises an eight-stranded beta-sandwich with two structural types as if the result of a circular permutation. Combining sequence, structural and modelling information, we have explored, at different levels of granularity, the functional characteristics of several families of C2 domains. At the coarsest level, the similarity correlates with key structural determinants of the C2 domain fold and, at the finest level, with the domain architecture of the proteins containing them, highlighting the functional diversity between the various sub-families. The functional diversity appears as different conserved surface patches throughout this common fold. In some cases, these patches are related to substrate-binding sites whereas in others they correspond to interfaces of presumably permanent interaction between other domains within the same polypeptide chain. For those related to substrate-binding sites, the predictions overlap with biochemical data in addition to providing some novel observations. For those acting as protein-protein interfaces, our modelling analysis suggests that slight variations between families are a result of not only complementary adaptations in the interfaces involved but also different domain architecture. In the light of the sequence and structural genomic projects, the work presented here shows that modelling approaches along with careful sub-typing of protein families will be a powerful combination for a broader coverage in proteomics. PMID- 14556750 TI - The relationship between conservation, thermodynamic stability, and function in the SH3 domain hydrophobic core. AB - To investigate the relationships between sequence conservation, protein stability, and protein function, we have measured the thermodynamic stability, folding kinetics, and in vitro peptide-binding activity of a large number of single-site substitutions in the hydrophobic core of the Fyn SH3 domain. Comparison of these data to that derived from an analysis of a large alignment of SH3 domain sequences revealed a very good correlation between the distinct pattern of conservation observed at each core position and the thermodynamic stability of mutants. Conservation was also found to correlate well with the unfolding rates of mutants, but not to the folding rates, suggesting that evolution selects more strongly for optimal native state packing interactions than for maximal folding rates. Structural analysis suggests that residue-residue core packing interactions are very similar in all SH3 domains, which provides an explanation for the correlation between conservation and mutant stability effects studied in a single SH3 domain. We also demonstrate a correlation between stability and the in vivo activity of mutants, and between conservation and activity. However, the relationship between conservation and activity was very strong only for the three most conserved hydrophobic core positions. The weaker correlation between activity and conservation seen at the other seven core positions indicates that maintenance of protein stability is the dominant selective pressure at these positions. In general, the pattern of conservation at hydrophobic core positions appears to arise from conserved packing constraints, and can be effectively utilized to predict the destabilizing effects of amino acid substitutions. PMID- 14556751 TI - Crystal structure of the extracellular protein secretion NTPase EpsE of Vibrio cholerae. AB - Type II secretion systems consist of an assembly of 12-15 Gsp proteins responsible for transporting a variety of virulence factors across the outer membrane in several pathogenic bacteria. In Vibrio cholerae, the major virulence factor cholera toxin is secreted by the Eps Type II secretion apparatus consisting of 14 Eps proteins. One of these, EpsE, is a cytoplasmic putative NTPase essential for the functioning of the Eps system and member of the GspE subfamily of Type II secretion ATPases. The crystal structure of a truncated form of EpsE in nucleotide-liganded and unliganded state has been determined, and reveals a two-domain architecture with the four characteristic sequence "boxes" of the GspE subfamily clustering around the nucleotide-binding site of the C domain. This domain contains two C-terminal subdomains not reported before in this superfamily of NTPases. One of these subdomains contains a four-cysteine motif that appears to be involved in metal binding as revealed by anomalous difference density. The EpsE subunits form a right-handed helical arrangement in the crystal with extensive and conserved contacts between the C and N domains of neighboring subunits. Combining the most conserved interface with the quaternary structure of the C domain in a distant homolog, a hexameric model for EpsE is proposed which may reflect the assembly of this critical protein in the Type II secretion system. The nucleotide ligand contacts both domains in this model. The N2-domain-containing surface of the hexamer appears to be highly conserved in the GspE family and most likely faces the inner membrane interacting with other members of the Eps system. PMID- 14556752 TI - Extended-spectrum beta-lactamases: implications for the clinical microbiology laboratory, therapy, and infection control. AB - Extended-spectrum beta-lactamase (ESBL) producing gram-negative bacilli are a growing concern in human medicine today. When producing these enzymes, organisms (mostly K. pneumoniae and E. coli) become highly efficient at inactivating the newer third-generation cephaloporins (such as cefotaxime, ceftazidime, and ceftriaxone). In addition, ESBL-producing bacteria are frequently resistant to many classes of non-beta-lactam antibiotics, resulting in difficult-to-treat infections. This review gives an introduction into the topic and is focused on various aspects of ESBLs; it covers the current epidemiology, the problems of ESBL detection and the clinical relevance of infections caused by ESBL-producing organisms. Therapeutic options and potential strategies for dealing with this growing problem are also discussed in this article. PMID- 14556753 TI - Circulating levels of granulocyte macrophage colony-stimulating factor in patients with the systemic inflammatory response syndrome. AB - OBJECTIVES: Granulocyte-macrophage colony stimulating factor (GM-CSF) is a key regulator cytokine that modulates the proliferation and maturation of polymorphonuclear and mononuclear progenitors. This study was designed to investigate and clarify the role of GM-CSF in 52 critically ill patients with systemic inflammatory response syndrome (SIRS). METHODS: Serum levels of GM-CSF were detected by an immunoenzyme assay. RESULTS: Our results clearly show that the serum concentrations of GM-CSF were significantly elevated in patients with infectious and noninfectious SIRS (33.2+/-45.7pg/ml, controls: 17.2+/-9.8pg/ml; p=0.0303). In addition, GM-CSF levels significantly decreased in patients with SIRS, particularly in patients with infectious SIRS, 5 and 7 days later. There was a clear tendency toward higher levels of GM-CSF in patients with poor, as compared with those having a good outcome of the disease. CONCLUSION: These results show that GM-CSF may play an important role in patients with infectious and noninfectious SIRS, and that GM-CSF levels progressively and significantly decrease in patients with infectious SIRS. PMID- 14556754 TI - Clinical prognostic markers in patients with severe sepsis: a prospective analysis of 139 consecutive cases. AB - OBJECTIVES: To analyze the clinical characteristics and determine predictive factors of mortality in previously healthy individuals suffering from severe sepsis. METHODS: The study included 139 patients with severe sepsis, admitted to the Department of Medicine over a two years period. Data recorded on admission included demographic information, blood pressure, core temperature, white blood count, hepatic and renal function tests, coagulation factors, blood gases, serum lactic acid levels, simplified acute physiology score (SAPS-II) and Glasgow Coma Scale (GCS). RESULTS: On admission, 62 patients were hypotensive, 52 had signs of diffuse intravascular coagulation (DIC), 72 had renal and 27 hepatic dysfunction. The overall mortality rate was 27.3%. Twenty-nine patients had septic shock on admission with a mortality rate of 62.07%. Hypoxemia, metabolic acidosis and the presence of DIC were more frequent in non-survivors, who also had significantly higher SAPS-II on admission and days 3 and 7. Independent factors associated with mortality were older age, septic shock, DIC and acute renal failure on admission, as well as SAPS-II at all time points and lactic acid levels on day 7. CONCLUSIONS: Septic patients with advanced age, septic shock, renal failure, DIC and metabolic acidosis on admission are at increased risk of mortality. The sustained presence of high SAPS-II and lactacidemia one week after admission are also important risk factors of poor outcome. PMID- 14556755 TI - Are infections due to resistant pathogens associated with a worse outcome in critically ill patients? AB - OBJECTIVES: To evaluate the outcome of critically ill patients infected with antimicrobial resistant microorganisms, and to analyse the factors involved in the development of antimicrobial resistance. METHODS: All patients admitted to a 31-bed mixed medico-surgical intensive care unit who developed a nosocomial infection were prospectively followed until discharge or death. RESULTS: Of 949 consecutive patients admitted, 186 developed a nosocomial infection: 79 with an antimicrobial-resistant pathogen and 107 with susceptible strains. The lungs were the main source of infections in both groups. The main resistant microorganisms were Enterobacter aerogenes, methicillin resistant Staphylococcus aureus (MRSA), and Enterobacter cloacae. The main susceptible microorganisms were Enterobacter spp., methicillin susceptible S. aureus (MSSA), and Proteus mirabilis. Patients infected with resistant strains had a longer length of stay prior to infection (9+/-4 vs. 5+/-3 days), longer total length of stay (18+/-16 vs. 11+/-7 days), longer duration of mechanical ventilation (12+/-15 vs. 6+/-7 days), and more severe coagulation, liver, and renal dysfunction (all p<0.05). The maximum degrees of organ failure during the ICU stay, and the respiratory dysfunction, but not infection with a resistant pathogen, were independent predictors for death. Multivariate logistic regression revealed previous use of multiple antibiotics, duration of length of stay prior to infection, and the degree of liver failure as independent factors for development of infection with resistant organisms. CONCLUSIONS: Infection with antimicrobial resistant microorganisms is not an independent predictor for death. The development of antimicrobial resistance is related to the previous use of multiple antibiotics, the ICU length of stay, and the severity of hepatic dysfunction. PMID- 14556757 TI - Provision of hepatitis B vaccination for primary care dental staff in Scotland. AB - OBJECTIVES: All dental surgeons should be protected from hepatitis B virus (HBV) infection by immunisation, ideally administered and monitored via occupational health services (OHS). This study examined relevant OHS systems in place for dental primary care healthcare workers (DHCW) across all Health Board Areas (HBAs) in Scotland. It also explored the DHCWs' knowledge of, and access to, these systems in three HBAs. METHODS: Data from senior staff in all Scottish Health Boards and Primary Care Trusts were collected by self-completing questionnaires. Information from DHCWs was collected via telephone interviews with General Dental Practitioners (GDPs) and Community Dental Officers (CDOs) in each of Ayrshire and Arran, Highland and Lothian Health Boards. RESULTS: Thirteen of the 15 HBAs had robust HBV vaccination and monitoring systems. However, only 7/15 (47%) of these covered all DHCWs. Seven HBAs provided vaccination and monitoring for CDOs only, leaving GDPs to undertake these responsibilities for themselves. Of the 105 DHCWs approached, 82 gave an interview. These interviews highlighted major differences between HBAs in relation to access of DHCWs to OHS and indicated that CDOs had greater access than GDPs to OHS. Overall, 31% of DHCWs were not satisfied with the OHS available. CONCLUSION: In order to safeguard both staff and patients, significant further work is required to ensure that all DHCWs have access to appropriate OHS support for provision and monitoring of immunisation procedures and related functions such as management of sharps injuries. PMID- 14556756 TI - The role of induced sputum in the diagnosis of pulmonary tuberculosis. AB - OBJECTIVES: To determine the sensitivity of referral guidelines for induced sputum in pulmonary tuberculosis (PTB) case detection and determine the diagnostic yield of the test. METHODS: A retrospective analysis of case notes of adult patients referred for induced sputum was carried out at the North Trent Regional Department of Infection and Tropical Medicine, the Royal Hallamshire Hospital, Sheffield, UK. Clinical characteristics were noted on patients referred during a 12-month period in 2001-2002, and of all patients with positive induced sputum results from April 1998 to March 2002. RESULTS: Over a 12-month period 114 patients were referred in our unit for induced sputum sampling. Twelve of the 114 patients had positive sputum cultures giving a positive yield for the test of 10.5%. With strict adherence to our referral guidelines, 10 of the 12 positive cases (83.3%) would have been detected. With modification to the referral guidelines, all 12 microbiologically positive cases would have been detected making the guidelines 100% sensitive in detecting microbiologically positive cases among patients referred with a clinical suspicion of PTB who are unable to self-expectorate. The positive yield for the test would have been increased to 13.2%. In the past 4 years 29 patients had positive induced sputum results. Of these, 11 patients (37.9%) had concurrent microbiological proven extra-pulmonary TB (EPTB) and 10 (34.5%) had past histories of TB infection. Seven patients had chest X-rays reported as normal. CONCLUSIONS: Induced sputum offers a relatively non-invasive investigation for patients with suspected PTB who are unable to bring up sputum. Induced sputum also offers a means of establishing a microbiological diagnosis in some cases of EPTB. An investigative algorithm is suggested.Summary. In an era of increasing drug resistant disease, establishing a microbiological diagnosis of TB is paramount. Sputum induction can be used as an alternative investigation to bronchoscopy in suspected PTB patients who are able to self-expectorate. Induced sputum is a sensitive investigation for suspected PTB patients but with a low diagnostic yield. Referral guidelines are suggested. PMID- 14556758 TI - Ileitis due to Yersinia pseudotuberculosis in Crohn's disease. AB - We present the case of a young woman with fulminating terminal ileitis due to Yersinia pseudotuberculosis, in whom Crohn's disease was subsequently diagnosed. Serological testing was prompted by radiological, macroscopic and histological changes which were atypical of Crohn's disease. We speculate that Y. pseudotuberculosis ileitis may occasionally produce an acute exacerbation of Crohn's disease, but that its occurrence may not be diagnosed if the atypical features are not recognised. In this case, the classical features of Y. pseudotuberculosis may have been modified by pre-existing Crohn's disease. We propose that, because of defects in mucosal barrier function and immunological function, patients with Crohn's may have increased susceptibility to infection by Yersinia sp. PMID- 14556759 TI - Necrotizing granulomatous vasculitis in advanced HIV infection. AB - We describe the first case of granulomatous necrotizing vasculitis not restricted to the central nervous system in an HIV-infected patient. No mycobacteria or drugs potentially associated with granuloma formation were involved in this patient, suggesting that the cause of this vasculitis was probably autoimmune. The development of granulomatous vascular inflammation in this patient with less than 200 CD4 cells/microl might have been due to immune overactivation. After starting potent antiretroviral therapy a profound immune deactivation was observed and the vasculitis did not relapse. PMID- 14556760 TI - Aspergillus endophthalmitis: an unusual complication of disseminated infection in renal transplant patients. AB - Endogenous Aspergillus endophthalmitis (AE) is a rare complication of invasive aspergillosis (IA) in transplant patients. In this report, we describe two patients with polycystic kidney disease, who developed AE with cerebral involvement after renal transplantation. Both patients received intense immunosuppression with methyl prednisolone and mycophenolate mofitil (MMF) because of persistent rejection, which rendered them diabetic and vulnerable to opportunistic infections. Endophthalmitis developed within six months of transplantation and was confirmed by microscopy and culture of the vitreous fluid. Patients were treated with combinations of different anti-fungal agents including liposomal amphotericin B, 5-flucytosine, itraconazole, voriconazole and terbinafine. In an electronic MEDLINE review, we found eight further cases of AE in renal transplant patients between 1959 and September 2002. Based on this review, we identified possible risk factors including CMV infection, diabetes mellitus and treatment for rejection with agents such as methyl prednisolone and MMF. In 70% of cases the histology, microscopy or culture of vitreous fluid confirmed the diagnosis. The outcome of AE in renal transplant patients was poor with 70-100% mortality. The review of reported cases and current practice guidelines suggests that vitrectomy and intravitreal amphoterecin B is the treatment of choice. In addition, new antifungal agents with good CSF and ocular penetration such as voriconalzole should be considered for the treatment of invasive cerebral/ocular aspergillosis. PMID- 14556761 TI - Polymerase chain reaction aids in the diagnosis of an unusual case of Aspergillus niger endocarditis in a patient with acute myeloid leukaemia. AB - Endocarditis secondary to Aspergillus niger has not been described in a leukaemic patient. We describe a case of A. niger endocarditis in a patient with acute myeloid leukaemia and refractory fever. The microbiological cause of his endocarditis was initially misdiagnosed because he fulfilled the Duke criteria for enterococcal endocarditis. A polymerase chain reaction test utilizing pan fungal primers detected a product from an Aspergillus sp. The DNA was subsequently sequenced and was found to have 100% homology with A. niger. A postmortem revealed fungal endocarditis secondary to disseminated aspergillosis, without evidence of bacterial endocarditis. The patient was found to have a lung aspergilloma that was possibly occupationally acquired, and may have been long standing. PMID- 14556763 TI - Sir, an offal blackening! PMID- 14556762 TI - Successful treatment of Xylohypha bantiana brain abscess mimicking invasive cerebral aspergillosis in a liver transplant recipient. AB - Invasive aspergillosis is a highly lethal disease in immunocompromised hosts especially with central nervous system involvement. In a review of 2380 liver transplants performed over 14 years, 13 out of 15 fungal brain abscesses were due to Aspergillus species and only one patient survived [Arch Surg, 132 (1997) 304]. Dematiaceous fungi while less readily recognised, often have a more favorable outcome [Transplantation, 63 (1997) 160]. We report a Xylophypha bantiana cerebral abscess in a liver transplant recipient initially identified as a branching mould on histology that had a successful outcome due to aggressive appropriate treatment. PMID- 14556765 TI - Energy medicine united. PMID- 14556766 TI - The use of herbs by pregnant and childbearing women: a risk-benefit assessment. AB - This paper reviews some of the evidence in relation to herbal remedies, and considers both their efficacy and safety for pregnant and childbearing women. It concludes with some suggested guidelines for midwifery and obstetric practice. PMID- 14556767 TI - Twin quilts--stories to be told. AB - A perusal of the literature reveals the many ways quilt design and personal communication is interwoven. We selected quilt making to provide those affected by breast cancer-as a patient, friend or relative-the opportunity to communicate their experience through quilt making. With the participation of community quilters, a concept and quilt design was developed to incorporate the work of patients and others in making a quilt. The project pattern and materials were presented. Contributors were invited to write about what inspired them to participate. For the 80 blocks that were constructed as twin quilts, 36 individuals wrote something about their block. Analysis of the vignettes identified major categories. The most common theme was honoring a specific person's life and their breast cancer journey. Displayed together with their accompanying vignettes, the quilts have become a focal point for interaction among patients and their families. PMID- 14556768 TI - Health hazards and electromagnetic fields. AB - Biological rhythms, physical wellbeing and mental states are dependent on our electrical brainwave system interacting with the extremely weak electromagnetic fields generated by the Earth's telluric and Cosmic radiations. In a single generation, since the evolution of humankind over millions of years, we are exposed to a wide range of powerful, artificially generated electromagnetic radiation which adversely affects the subtle balance in nature's energy fields and has become the source of so-called 'diseases of civilization'. This also includes electromagnetic sensitivity. Generally, there is a lack of awareness and understanding of the impact electromagnetic fields can have upon health and wellbeing.Our ancestors were acutely aware that certain locations, were perceived to have a positive energy field which was beneficial to health and vitality. Over time, these areas are now referred to as sacred sites for spiritual ceremony and as healing centres. In contrast, there are other geographical locations that can have a negative effect upon health and these are known as geopathic stress zones. It is believed that such zones can interfere with the brain's normal function that inhibits the release of melatonin and other endocrine secretions needed to replenish the immune system. Geopathic stress can affect animals and plant life as well as human beings and significantly contributes to sick building syndrome (SBS). Whilst there is an increasing body of opinion amongst eminent researchers and scientists who are addressing these issues, the establishment professions are slow to change. However, very gradually, modern allopathic medicine and attitudes are beginning to recognise the extraordinary wisdom and efficacy of ancient traditions such as acupuncture, light, colour and other therapies based on the understanding and treatment of the interaction of a person's electromagnetic subtle body and the immediate environment. These and many other 'complementary' therapies may soon become mainstream medical practice. In the meantime, we can help ourselves by learning how to detect the hazards and daily practise prudent avoidance. PMID- 14556769 TI - Qigong: harmonising the breath, the body and the mind. AB - Qigong is a form of exercise developed in China over millennia to prevent illness and foster physical and mental health. Despite many claims of the therapeutic effects of Qigong, the fundamental concepts and principles underlying this holistic therapy remain unfamiliar to Western populations. This article helps practitioners to understand the art of Qigong based on three main principles, harmonising the breath, the body and the mind. PMID- 14556770 TI - Single-blind trial addressing the differential effects of two reflexology techniques versus rest, on ankle and foot oedema in late pregnancy. AB - This single-blind randomised controlled trial explored the differential effects of two different foot reflexology techniques with a period of rest on oedema relieving effects and symptom relief in healthy pregnant women with foot oedema. Fifty-five women in the third trimester were randomly assigned to one of the three groups: a period of rest, 'relaxing' reflexology techniques or a specific 'lymphatic' reflexology technique for 15 min with pre- and post-therapy ankle and foot circumference measurements and participant questionnaire. There was no statistically significant difference in the circumference measurements between the three groups; however, the lymphatic technique reflexology group mean circumference measurements were all decreased. A significant reduction in the women's symptom mean measurements in all groups (p<0.0001) was apparent. A 'perceived wellbeing' score revealed the lymphatic technique group (p<0.0001) significantly increased their wellbeing the most, followed closely by relaxing techniques (p<0.001) and then the control rest group (p<0.03). Lymphatic reflexology techniques, relaxing reflexology techniques and a period of rest had a non-significant oedema-relieving effect. From the women's viewpoint, lymphatic reflexology was the preferred therapy with significant increase in symptom relief. PMID- 14556771 TI - The lecture from hell. PMID- 14556772 TI - Return to TRI: Winston Churchill Fellow and Salford University Scholar travel to Miami and the Keys. AB - During May and June 2003, the 2 authors visited the internationally renowned Touch Research Institute based at the University of Miami and Jackson Memorial Hospital. For one of the authors, Peter Mackereth, this was a return visit. Both authors are Ph.D. students who have successfully obtained travel scholarships to participate in seminars and laboratory work. The paper will report on the experience of their visit, the current and future work of the TRI and its collaborators. Jacqui took the opportunity to spend time at the Dolphin Research Centre, this provided insight into the therapeutic value of interacting with these amazing mammals. Peter also took the opportunity to visit a group of massage therapists working in Key West. PMID- 14556773 TI - Immunobiology of acute graft-versus-host disease. AB - Graft-versus-host disease (GVHD) has been the primary limitation to the wider application of allogeneic bone marrow transplantation (BMT). The immunobiology of acute GVHD is complex and can be conceptualized to be a three-step process. In step 1, the conditioning regimen (irradiation and/or chemotherapy) leads to the damage and activation of host tissues and induces the secretion of inflammatory cytokines TNF-alpha and IL-1. As a consequence expression of MHC antigens and adhesion molecules is increased, thus enhancing the recognition of host alloantigens by donor T cells. Donor T-cell activation in step 2 is characterized by donor T-cell interaction with host APCs and subsequent proliferation, differentiation, and secretion of cytokines. Cytokines such as IL-2 and IFN-gamma enhance T-cell expansion, induce cytotoxic T cells (CTL) and natural killer (NK) cell responses, and prime additional mononuclear phagocytes to produce TNF-alpha and IL-1. These inflammatory cytokines in turn stimulate production of inflammatory chemokines, thus recruiting effector cells into target organs. In step 3, effector functions of mononuclear phagocytes are triggered via a secondary signal provided by lipopolysaccharide (LPS) that leaks through the intestinal mucosa damaged during step 1. This mechanism may result in the amplification of local tissue injury and further promotion of an inflammatory response, which, together with the CTL and NK components, leads to target tissue destruction in the transplant host. PMID- 14556774 TI - Anaemia and red blood cell transfusion in the critically ill patient. AB - Anaemia is a common finding in critically ill patients. There are often multiple causes. Obvious causes include surgical bleeding and gastrointestinal haemorrhage but many patients have no overt bleeding episodes. Phlebotomy can be a significant source of blood loss. In addition, critically ill patients have impaired erythropoiesis as a consequence of blunted erythropoietin production and direct inhibitory effects of inflammatory cytokines. The ability of a patient to tolerate anaemia depends on their clinical condition and the presence of any significant co-morbidity; maintenance of circulating volume is of paramount importance. There is no universal transfusion trigger. Current guidelines for critically ill and perioperative patients advise that at Hb values <70 g/L red blood cell transfusion is strongly indicated and at Hb values >100 g/L transfusion is unjustified. For patients with Hb values in the range 70 to 100 g/L the transfusion trigger should be based on clinical indicators. Most stable critically ill patients can probably be managed with a Hb concentration between 70 and 90 g/L. Uncertainties exist concerning the most appropriate Hb concentration for patients with significant cardio-respiratory disease. PMID- 14556775 TI - Congenital neutropenia. AB - Congenital neutropenia is strictly defined as neutropenia present at birth. However, it is more generally used to describe neutropenia secondary to inherited genetic mutations. This review will discuss the presentation of such children and the various causes of congenital neutropenia. In particular, it will focus on severe congenital neutropenia (SCN) and the recent discovery of mutations in the gene encoding neutrophil elastase in the majority of cases of SCN. The potential mechanisms of pathogenesis and of transformation to leukaemia will be discussed. Shwachman-Diamond Syndrome and other less common causes of congenital neutropenia will also be reviewed. Finally, an approach to the child with potential congenital neutropenia will be presented. PMID- 14556776 TI - Dyskeratosis congenita: its link to telomerase and aplastic anaemia. AB - Dyskeratosis congenita (DC) is an inherited bone marrow failure syndrome exhibiting considerable clinical and genetic heterogeneity. X-linked recessive, autosomal dominant and autosomal recessive forms are recognised. The gene mutated in X-linked DC (DKC1) encodes a highly conserved nucleolar protein called dyskerin. Dyskerin associates with the H/ACA class of small nucleolar RNAs which are important in guiding the conversion of uracil to pseudouracil in ribosomal RNA. Dyskerin also associates with the RNA component of telomerase (hTR) which is important in the maintenance of telomeres. Mutations in hTR were recently demonstrated in patients with autosomal dominant DC and in a subset of patients with aplastic anaemia (AA) but without other diagnostic features of DC. This discovery demonstrates that both DC and a subset of AA are due to a defect in telomerase. The link between DC and AA and in turn to defective telomerase suggests that treatments directed at correction of telomerase activity might benefit DC/AA patients who do not respond to conventional therapy. PMID- 14556777 TI - Stem cells and immune reconstitution in AIDS. AB - The hematopoietic system generally has reserve sufficient to tolerate significant insult and regenerative capacity to overcome most damage due to infectious agents. However, HIV infection results in a progressive decline in hematopoietic function and even in the context of potent, anti-retroviral therapy is able to only incompletely reconstitute immune function. The ability of the immune system to respond to HIV itself remains compromised, a defect that leaves infected individuals with a lifelong dependence on medications. The capability of stem cells and the thymus to restore function and their limitations in the context of HIV infection are discussed in this review. PMID- 14556778 TI - Stem cell transplantation for multiple sclerosis: what is the evidence? AB - Experimental and clinical observations have indicated that high-dose immunosuppression followed by autologous stem cell transplantation (ASCT) can induce remissions in severe, refractory, autoimmune diseases including multiple sclerosis (MS), a T cell-mediated autoimmune disorder against CNS myelin components, causing severe chronic disability. Control of the disease is unsatisfactory in most of the patients, especially those with rapidly evolving relapsing-remitting course and those with chronic progressive disease. The rationale for treating autoimmune diseases with ASCT is based on the immunosuppressive and immunomodulating effects of ASCT which may shift the immunological balance towards disease quiescence, a hypothesis supported by the results of ASCT in animal models of MS and by clinical observations in MS patients transplanted for concurrent malignancies. A number of phase I-II studies of ASCT in patients with active MS, conducted worldwide since 1995, and a comprehensive analysis of 85 patients, recently reported by the European Group for Blood and Marrow Transplantation (EBMT), have shown the feasibility of the method, a prominent anti-inflammatory effect on magnetic resonance imaging (MRI) disease, and a possible clinical benefit for active and refractory cases. The impact on MRI disease parameters appears superior with ASCT than with conventional therapies but the clinical results, in terms of stabilization of disease and prevention of disability, need to be validated in prospective, controlled trials. The procedure is also associated with a transplant-related mortality risk, of about 5% in high-risk cases, i.e., in older patients, those with high disability scores, those receiving strong myeloablative conditioning regimens and those undergoing intensive in vivo or ex vivo T cell-depletion. Therefore, it could be recommended for the treatment of a chronic, non-lethal, disease like MS only if it proved superior to standard therapies. A randomized trial is now launched by the EBMT to compare ASCT to mitoxantrone, currently regarded as one of the best available treatments, in properly selected patients having high chance of response at minimal mortality risk. PMID- 14556779 TI - Receptor tyrosine kinases in normal and malignant haematopoiesis. AB - Haematopoiesis is controlled by a number of growth factors and cytokines, a number of which act through binding to high-affinity receptor tyrosine kinases (RTKs). Approximately 20 different RTK classes have been identified, all of which share a similar structure that includes a ligand binding extracellular domain, a single transmembrane domain and an intracellular tyrosine kinase domain. Recent studies have linked an increasing number of mutations in the RTKs to the pathogenesis of both acute and chronic leukaemia. For example, the FLT3 receptor, a RTK class III, is the most commonly mutated gene in acute myeloid leukaemia, while c-kit mutations are strongly linked to the development of mast cell malignancy. This review summarizes the RTK classes that are known to be expressed on normal haematopoietic tissue and highlights the many 'gain-of-function' mutations involved in leukaemogenesis. It is to be hoped that this knowledge will provide important new insights for targeted therapy in leukaemia. PMID- 14556780 TI - Treatment of human immunodeficiency virus-related lymphoma with haematopoietic stem cell transplantation. AB - The advent of highly active antiretroviral therapy (HAART) and its co administration with chemotherapy in patients with human immunodeficiency virus (HIV)-related lymphoma has lead to the exploration of potentially curative combination chemotherapy and myeloablative therapy followed by autologous haematopoietic stem cell transplantation (ASCT). Applying the same principles used for patients with HIV-negative aggressive lymphoma, in 1998 we developed a program of high-dose therapy and ASCT at City of Hope for patients with HIV related lymphoma and Hodgkin's disease. Our studies have primarily included patients with chemosensitive lymphoma in relapse or first remission with poor risk features at diagnosis. Filgrastim (G-CSF)-primed peripheral blood stem cell mobilization and apheresis have been successful while patients were receiving HAART and chemotherapy. To date, ASCT has been performed in 19 patients with HIV related lymphoid malignancies, representing the largest single-institution experience reported to date. Most patients received a chemotherapy-based conditioning regimen consisting of high-dose carmustine, etoposide and cyclophosphamide. Early infections, namely bacteremias and neutropenic fever were similar to those observed in the HIV-negative transplant setting. Opportunistic infections were rare and easily treatable. There were three early deaths, two from relapsed lymphoma and one from multi-organ failure in an older patient. The remaining 16 patients are alive and in remission. In summary, ASCT is well tolerated, can result in long-term remissions, and is potentially curative in selected HIV-related lymphoma patients with chemosensitive relapse and high-risk disease in first remission defined by the age-adjusted International Prognostic Index criteria (i.e., two or three of the following: elevated LDH, advanced stage, and poor performance status). Acquisition of resistance to HAART remains as a potential problem for HIV-positive patients who are cured of their lymphoma. PMID- 14556781 TI - Human cytomegalovirus-specific immunity following haemopoietic stem cell transplantation. AB - The herpesvirus Human Cytomegalovirus (HCMV) is an important opportunistic infection in recipients of allogeneic haemopoietic stem cell transplants, in whom HCMV-specific CD8+ and CD4+ T-cell responses are impaired. The nature of the HCMV specific T-cell response in healthy virus carriers has been characterised in detail. High frequencies of circulating CD8+ T-cells that recognise defined viral peptides are maintained for years, and include individual CD8+ clones that have undergone extensive clonal expansion and phenotypic diversification in vivo. Following stem cell transplantation, the kinetics of HCMV-specific CD8+ T-cell reconstitution in the recipient are related to the presence or absence of antigen experienced CD8+ T-cells transferred via the allograft, and to the presence of the virus in the recipient. We discuss recent progress in our understanding of HCMV-specific immunity in healthy virus carriers and in recipients after alloSCT. PMID- 14556784 TI - Synthesis and evaluation of alpha,alpha-disubstituted-3-mercaptopropanoic acids as inhibitors for carboxypeptidase A and implications with respect to enzyme inhibitor design. AB - 2-Ethyl-2-methyl-3-mercaptopropanoic acid (6) and 2-benzyl-2-methyl-3 mercaptopropanoic acid (7) were synthesized and evaluated as inhibitors for carboxypeptidase A (CPA), a prototypical zinc protease with the expectation that the binding affinities of these inhibitors would be augmented over those of 2 ethyl-3-methylsuccinic acid (2) and 2-benzyl-3-methylsuccinic acid (3), respectively, in light of the fact that the sulfhydryl group is a better zinc coordinating moiety than the carboxylate group. Contrary to the expectation, however, the inhibitory potency of 6 was not improved and that of 7 was rather attenuated by the replacement. A probable explanation for the unexpected results is offered. PMID- 14556785 TI - 2,4-Diaminopyrimidines as inhibitors of Leishmanial and Trypanosomal dihydrofolate reductase. AB - This paper describes the synthesis of 4'-substituted and 3',4'-disubstituted 5 benzyl-2,4-diaminopyrimidines as selective inhibitors of leishmanial and trypanosomal dihydrofolate reductase. Compounds were then assayed against the recombinant parasite and human enzymes. Some of the compounds showed good activity. They were also tested against the intact parasites using in vitro assays. Good activity was found against Trypanosoma cruzi, moderate activity against Trypanosoma brucei and Leishmania donovani. Molecular modeling was undertaken to explain the results. The leishmanial enzyme was found to have a more extensive lipophilic binding region in the active site than the human enzyme. Compounds which bound within the pocket showed the highest selectivity. PMID- 14556786 TI - Studies on Quinones. Part 38: synthesis and leishmanicidal activity of sesquiterpene 1,4-Quinones. AB - The reaction of (+)-euryfuran 1 with several benzo-, naphtho- and benzo[b]thiophene-1,4-quinones in acetic acid yields the corresponding euryfuryl 1,4-quinones 3, 5, 7, 8, 10, 12, and 14. The structure of compounds 7, 8, 12, and 14 was assigned through 2D NMR 1H-13C HMBC experiments. The influence of the acidity of the solvent upon the reactivity and regioselectivity of the quinones to the oxidative coupling reaction, is discussed. The in vitro activity of the euryfurylquinones and their corresponding precursors against Leishmania amazonensis is described. PMID- 14556787 TI - Small hydroxyethylene-based peptidomimetics inhibiting both HIV-1 and C. albicans aspartic proteases. AB - We have extended a highly flexible method for rapidly assembling aspartic protease inhibitors to produce symmetric and asymmetric monohydroxyethylene peptidomimetics. This method is based on the prior synthesis of the central non cleavable peptide-bond isostere [NH(2)-P(1)psiP1'-NH(2); psi=hydroxyethylene isostere, HNCH(Bz)CHOHCH(2)CH(Bz)NH], with the possibility of accurately controlling its stereochemistry (S,S,S or S,R,S), and subsequently adding appropriate flanking units, chosen from commercially available amino acids, aromatic carboxylic acids, or phenoxyacetic acid (Poa) derivatives. The method was used to make asymmetric inhibitors of general formula Kyn-Xaa-PhepsiPhe dmPoa, (Kyn=kynurenic acid, Xaa=Val, Thr or D-thienylglycine, M(r)=716-754) and symmetric inhibitors of formula xPoa-PhepsiPhe-xPoa (xPoa=Poa or dimethyl-, hydroxy-, formyl- or acetyl-Poa, M(r)=553-609), with logP(o/w) values ranging from 4.1 to 7.6. Inhibition of HIV-PR did not depend on the stereochemistry of the hydroxyl group, while it depended markedly on the substituents present on the Poa residues, with dmPoa being preferred over Poa or its more hydrophilic derivatives. Conversely, inhibition of Candida albicans Sap2 was higher for the S,S,S epimers, and Poa or its hydrophilic derivatives were preferred over dmPoa. PMID- 14556788 TI - Synthesis and structure-activity relationships of 5-phenylthiophenecarboxylic acid derivatives as antirheumatic agents. AB - 5-(Phenylthiophene)-3-carboxylic acid (2a), a metabolite of esonarimod (1), which was developed as a new antirheumatic drug, was considered as a lead compound for new antirheumatic drugs. A new series of 2a derivatives were synthesized and their characteristic pharmacological effects, that is their antagonistic effect toward interleukin (IL)-1 in mice and the suppressive effect against adjuvant induced arthritis (AIA) in rats, were evaluated and compared with those of 1. The structure-activity relationships indicated that [5-(4-bromophenyl)-thiophen-3 yl]acetic acid (5d), methyl [5-(4-chlorophenyl)-thiophen-3-yl]acetate (5h), and methyl [5-(4-bromophenyl)-thiophen-3-yl]acetate (5i) suppressed AIA more potently than 1 and all of the other synthesized compounds. PMID- 14556789 TI - Cochlioquinone A1, a new anti-angiogenic agent from Bipolaris zeicola. AB - Cochlioquinone A1 (CoA1) was newly isolated from the culture extract of Bipolaris zeicola as a potent anti-angiogenic agent. CoA1 inhibited in vitro angiogenesis of bovine aortic endothelial cells (BAECs) such as bFGF-induced tube formation and invasion at the concentration (1 microg/mL) without cytotoxicity. Notably, CoA1 exhibited more potent inhibition activity for the growth of BAECs than that of normal and cancer cell lines investigated in this study. These results demonstrate that CoA1 is a new anti-angiogenic agent and can be developed as a new therapeutic agent for angiogenesis-related diseases. PMID- 14556790 TI - Synthesis and pharmacological characterization of functionalized 2-pyridones structurally related to the cardiotonic agent milrinone. AB - A new class of cardiotonic agents characterized by a 2-pyridone structure was synthesized. Appropriate sym-2-dimethylaminomethylene-1,3-diones reacted with methylcyanoacetate to afford the desired compounds. These derivatives were evaluated for their ability in inducing cardiotonic response on guinea pig isolated myocardial preparations. Compound 8b increased atrial contractility to an extent which is significantly higher than that of milrinone, the parent drug used as a reference compound. The pharmacological characterization and the docking studies performed on 8b highlighted its selective mechanism of action via type 3 PDE (PDE3) inhibition. PMID- 14556791 TI - Diaryldimethylpiperazine ligands with mu- and delta-opioid receptor affinity: Synthesis of (+)-4-[(alphaR)-alpha-(4-allyl-(2S,5S)-dimethylpiperazin-1-yl)-(3 hydroxyphenyl)methyl]-N-ethyl-N-phenylbenzamide and (-)-4-[(alphaR)-alpha-(2S,5S) dimethylpiperazin-1-yl)-(3-hydroxyphenyl)methyl]-N-ethyl-N-phenylbenzamide. AB - We have explored the synthesis of compounds that have good affinity for both mu- and delta-opioid receptors from the (alphaR,2S,5S) class of diaryldimethylpiperazines. These non-selective compounds were related to opioids that have been found to interact selectively with mu- or delta-opioid receptors as agonists or antagonists. In our initial survey, we found two compounds, (+)-4 [(alphaR)-alpha-(4-allyl-(2S,5S)-dimethylpiperazin-1-yl)-(3-hydroxyphenyl)methyl] N-ethyl-N-phenylbenzamide (14) and its N-H relative, (-)-4-[(alphaR)-alpha (2S,5S)-dimethylpiperazin-1-yl)-(3-hydroxyphenyl)methyl]-N-ethyl-N phenylbenzamide (15), that interacted with delta-receptors with good affinity, and, as we hoped, with much higher affinity at mu-receptors than SNC80. The relative configuration of the benzylic position in (+)-4-[(alphaR)-alpha-(4-allyl (2S,5S)-dimethyl-1-piperazinyl)-(3-methoxyphenyl)methyl]-benzyl alcohol (10) was determined by X-ray crystallographic analysis of a crystal that was an unresolved twin. The absolute stereochemistry of that benzylic stereogenic center was unequivocally derived by the X-ray crystallographic analysis from the two other centers of asymmetry in the molecule that were known. Those were established from the synthesis via a dipeptide cyclo-L-Ala-L-Ala in which the absolute stereochemistry was established. PMID- 14556792 TI - Discovery of novel benzothiazolesulfonamides as potent inhibitors of HIV-1 protease. AB - The human immunodeficiency virus (HIV) has been shown to be the causative agent for AIDS. The HIV virus encodes for a unique aspartyl protease that is essential for the production of enzymes and proteins in the final stages of maturation. Protease inhibitors have been useful in combating the disease. The inhibitors incorporate a variety of isosteres including the hydroxyethylurea at the protease cleavage site. We have shown that the replacement of t-butylurea moiety by benzothiazolesulfonamide provided inhibitors with improved potency and antiviral activities. Some of the compounds have shown good oral bioavailability and half life in rats. The synthesis of benzothiazole derivatives led us to explore other heterocycles. During the course of our studies, we also developed an efficient synthesis of benzothiazole-6-sulfonic acid via a two-step procedure starting from sulfanilamide. PMID- 14556794 TI - Synthesis and biological evaluation of benzo[d]isothiazole, benzothiazole and thiazole Schiff bases. AB - Three new series of benzo[d]isothiazole, benzothiazole and thiazole Schiff bases were synthesized and tested in vitro with the aim of identifying novel lead compounds active against emergent and re-emergent human and cattle infectious diseases (AIDS, hepatitis B and C, tuberculosis, bovine viral diarrhoea) or against drug-resistant cancers (leukaemia, carcinoma, melanoma, MDR tumors) for which no definitive cure or efficacious vaccine is available at present. In particular, these compounds were evaluated in vitro against representatives of different virus classes, such as a HIV-1 (Retrovirus), a HBV (Hepadnavirus) and the single-stranded RNA(+) viruses Yellow fever virus (YFV) and Bovine viral diarrhoea virus (BVDV), both belonging to Flaviviridae. Title compounds were also tested against representatives of Gram-positive and Gram-negative bacteria (Staphylococcus aureus, Salmonella spp.), various atypic mycobacterial strains (Mycobacterium fortuitum and Mycobacterium smegmatis), yeast (Candida albicans) and mould (Aspergillus fumigatus). None of the compounds showed antiviral or antimicrobial activity. The benzo[d]isothiazole compounds showed a marked cytotoxicity (CC(50)=4-9 microM) against the human CD4(+) lymphocytes (MT-4) that were used to support HIV-1 growth. For this reason, the most cytotoxic compounds of this series were evaluated for their antiproliferative activity against a panel of human cell lines derived from haematological and solid tumors. The results highlighted that all the benzo[d]isothiazole derivatives inhibited the growth of leukaemia cell lines, whereas only one of the above mentioned compounds (1e) showed antiproliferative activity against two solid tumor-derived cell lines. PMID- 14556793 TI - Synthesis and in vitro evaluation of potential antichagasic hydroxymethylnitrofurazone (NFOH-121): a new nitrofurazone prodrug. AB - The synthesis of mutual prodrugs of nitrofurazone with primaquine, using specific and nonspecific spacer groups, has been previously attempted seeking selective antichagasic agents. The intermediate reaction product, hydroxymethylnitrofurazone (NFOH-121), was isolated and tested in LLC-MK(2) culture cells infected with trypomastigotes forms of Trypanosoma cruzi showing higher trypanocidal activity than nitrofurazone and benznidazol in all stages. The mutagenicity tests showed that the prodrug was less toxic than the parent drug. Degradation assays were carried out in pH 1.2 and 7.4. PMID- 14556795 TI - Synthesis and cytotoxicity evaluation of pyridin[2,3-f]indole-2,4,9-trione and benz[f]indole-2,4,9-trione derivatives. AB - 3-Ethoxycarbonyl-3-methyl-1N-substrituted-2,3-dihydro-pyridin[2,3-f]indole-2,4,9 trione [9(a-d)] and 3-ethoxycarbonyl-3-methyl-N-substrituted-2,3-dihydro benz[f]indole-2,4,9-trione [10(a-i)] derivatives were synthesized from 7-chloro-6 (1,1-diethoxycarbonyl-ethyl)-5,8-quinolinedione (7) and 2-chloro-3-(1,1 diethoxycarbonyl-ethyl)-1,4-naphthoquinone (8), respectively, using a variety of alkyl- and arylamines. The cytotoxic activities of the synthesized compounds were evaluated by a Sulforhodamine B (SRB) assay against the following tumor cell lines: A459 (human non-small cell lung), SK-OV-3 (human ovarian), SK-MEL-2 (human melanoma), XF498 (human CNS), and HCT 15 (human colon). Almost all the derivatives mentioned above had a more potent cytotoxic effect against SK-OV-3 than etoposide. In particular, 3-ethoxycarbonyl-3-methyl-N-(4-aminophenyl)-2,3 dihydro-benz[f]indole-2,4,9-trione (10h) exhibited greater activity against all the tumor cell lines, and its cytotoxic effect against SK-OV-3 was especially higher than doxorubicin. PMID- 14556796 TI - Oxidative conjugation of chlorogenic acid with glutathione. Structural characterization of addition products and a new nitrite-Promoted pathway. AB - Chlorogenic acid (1), a cancer chemopreventive agent widely found in fruits, tea and coffee, undergoes efficient conjugation with glutathione (GSH), in the presence of horseradish peroxidase/H(2)O(2) or tyrosinase at pH 7.4, to yield three main adducts that have been isolated and identified as 2-S glutathionylchlorogenic acid (3), 2,5-di-S-glutathionylchlorogenic acid (4) and 2,5,6-tri-S-glutathionylchlorogenic acid (5) by extensive NMR analysis. The same pattern of products could be obtained by reaction of 1 with GSH in the presence of nitrite ions in acetate buffer at pH 4. Mechanistic experiments suggested that oxidative conjugation reactions proceed by sequential nucleophilic attack of GSH on ortho-quinone intermediates. Overall, these results provide the first complete spectral characterization of the adducts generated by biomimetic oxidation of 1 in the presence of GSH, and disclose a new possible nitrite-mediated conjugation pathway of 1 with GSH at acidic pH of physiological relevance. PMID- 14556797 TI - Design, synthesis and pharmacological profile of novel dopamine D2 receptor ligands. AB - The present study describes the synthesis and pharmacological profile of three novel heterocyclic compounds originally designed, on the basis of bioisosterism, as dopamine D2 receptor ligands: 1-[1-(4-chlorophenyl)-1H-pyrazol-4-ylmethyl]-4 phenyl-piperazine (LASSBio-579), 1-phenyl-4-(1-phenyl-1H-[1,2,3]triazol-4 ylmethyl)-piperazine (LASSBio-580) and 1-[1-(4-chlorophenyl)-1H-[1,2,3]triazol-4 ylmethyl]-4-phenyl-piperazine (LASSBio-581). Binding studies performed on brain homogenate indicated that all three compounds bind selectively to D2 receptors. In addition, electrophysiological studies carried out in cultured hippocampal neurons suggested that LASSBio-579 and 581 act as D2 agonists, whereas LASSBio 580 acts as a D2 antagonist. PMID- 14556798 TI - Synthesis, molecular modeling and biological evaluation of aza-proline and aza pipecolic derivatives as FKBP12 ligands and their in vivo neuroprotective effects. AB - Nonimmunosuppressant ligands, exemplified by GPI 1046 (1), for the peptidyl prolyl isomerase FKBP12 have been found to unexpectedly possess powerful neuroprotective and neuroregenerative effects in vitro and in vivo. We have extensively explored the therapeutic utility of FKBP12 ligands based on analogues of proline and pipecolic acid. As part of our ongoing program to explore novel structural classes of FKBP12 ligands, we herein wish to report a new class of FKBP12 ligands containing aza-proline and aza-pipecolic acid analogues. Details of the synthetic studies, together with biological activity will be presented. PMID- 14556799 TI - Inhibitors of lipoprotein(a) assembly. AB - Compounds of the general structure A and B were investigated for their activity as lipoprotein(a), [Lp(a)], assembly (coupling) inhibitors. SAR around the amino acid derivatives (structure A) gave compound 14-6 as a potent coupling inhibitor. Oral dosing of compound 14-6 to Lp(a) transgenic mice and cymologous monkeys resulted in a>30% decrease in plasma Lp(a) levels after 1-2 weeks of treatment at 100 mg/kg/day. PMID- 14556800 TI - Is intracytoplasmic sperm injection safe? AB - OBJECTIVE: To review the pregnancy and birth outcomes of patients undergoing intracytoplasmic sperm injection (ICSI). DESIGN: Selective review of the literature. SETTING: Couples undergoing either conventional IVF or ICSI and their resulting offspring. PATIENT(S): None. MAIN OUTCOME MEASURE(S): None. INTERVENTION(S): None. RESULTS: These observational studies are unfortunately inherently biased. Regardless, the information and experiences from these large centers cannot be ignored. These experiences reveal that birth weight may be decreased by an amount that is generally not clinically relevant. Chromosomal and genetic abnormalities are increased probably only as a direct corollary to the underlying parental risk. Congenital malformations reveal no clustering of any single specific abnormality. Developmental assessment is available for up to 2 years, and no major delays have been identified in either motor or mental function. CONCLUSION(S): Intracytoplasmic sperm injection appears to be a safe alternative for couples who otherwise would be unable to achieve pregnancy. The inherent risks associated with these genetically "at risk" couples mandate thorough evaluation and counseling before undertaking ICSI. PMID- 14556801 TI - Preimplantation aneuploidy screening using comparative genomic hybridization or fluorescence in situ hybridization of embryos from patients with recurrent implantation failure. AB - OBJECTIVE: To select chromosomally euploid embryos for transfer by analyzing single biopsied blastomeres using either fluorescence in situ hybridization (FISH) for chromosomes 13, 16, 18, 21, and 22 or comparative genomic hybridization (CGH), which provides a full karyotype. DESIGN: Prospective observational study. SETTING: A large IVF unit and the research laboratory of a hospital clinical genetics unit. PATIENT(S): Twenty patients with recurrent implantation failure. INTERVENTION(S): Ovarian stimulation and IVF by intracytoplasmic sperm injection (ICSI), embryo biopsy, and embryo transfer. MAIN OUTCOME MEASURE(S): Chromosome normality of biopsied blastomeres and implantation and clinical pregnancy rates. RESULT(S): Comparative genomic hybridization was able to identify many chromosomal abnormalities that would have been missed if those cells had been analyzed by FISH. The clinical pregnancy rate per transfer and implantation rate was 11% and 7% for embryos analyzed by FISH and 21% and 15% for embryos analyzed by CGH. CONCLUSION(S): Comparative genomic hybridization is more effective than FISH for identifying chromosomally normal embryos, which may result in a higher clinical pregnancy rate and implantation rate after embryo transfer. PMID- 14556802 TI - Preimplantation diagnosis for aneuploidies using fluorescence in situ hybridization or comparative genomic hybridization. PMID- 14556803 TI - Questions concerning the suitability of comparative genomic hybridization for preimplantation genetic diagnosis. PMID- 14556804 TI - Aneuploidy screening of preimplantation embryos using comparative genomic hybridization versus fluorescence in situ hybridization techniques. PMID- 14556806 TI - Development of a nonsurgical diagnostic tool for endometriosis based on the detection of endometrial leukocyte subsets and serum CA-125 levels. AB - OBJECTIVE: To determine whether the proportion of several leukocyte subsets is modulated in the endometrium of patients with endometriosis and, if yes, whether it can be used for diagnostic purposes. DESIGN: Case-control study. SETTING: Eight clinical institutions of the Montreal area. PATIENT(S): Women who underwent laparoscopy or laparotomy between 1997 and 2001, who had regular menstrual cycles and were not under hormone treatment for the previous 3 months were selected. This study included 368 women, 173 with surgically confirmed endometriosis and 195 controls with no surgical evidence of endometriosis. INTERVENTIONS: MAIN OUTCOME MEASURE(S): Cytometry analysis was used to measure the proportion of several leukocyte subsets among CD45(+) endometrial cells. RESULT(S): The proportion of CD3(+), CD16(+), CD3(-)HLADR(-), CD3(-)CD45RA(-), CD3(+)CD16(-), CD3(+)CD56(-), CD56(-)CD16(+), and CD16b(+) leukocytes was significantly altered in the endometrium of cases compared with controls. A multiple logistic regression model was adjusted with these endometrial leukocytes, serum CA-125 levels, risk factors, and confounders. The diagnostic performance of this predictive model was defined by a specificity of 95% and a sensitivity of 61%. Furthermore, the positive and negative predictive values were 91% and 75%, respectively. CONCLUSION(S): This predictive model represents a novel diagnostic tool to identify women with a high likelihood of suffering from endometriosis. PMID- 14556807 TI - The elusive diagnostic test for endometriosis. PMID- 14556808 TI - Altered expression of interleukin-18 in the peritoneal fluid of women with endometriosis. AB - OBJECTIVE: Peritoneal fluid (PF) inflammatory factors may participate in the pathogenesis of endometriosis. The aim of this study was to investigate PF interleukin (IL)-18 levels in women with and without endometriosis. DESIGN: Controlled clinical study. SETTING: Women undergoing laparoscopy at a university hospital. PATIENT(S): Fifty women with previously untreated endometriosis, 8 women on GnRH agonists for endometriosis, and 18 control women with normal pelvic anatomy who were undergoing tubal ligation. INTERVENTION(S): Peritoneal fluid IL 18 levels as measured by ELISA. MAIN OUTCOME MEASURE(S): Peritoneal fluid IL-18 levels. RESULT(S): Peritoneal fluid IL-18 levels were significantly higher in women with previously untreated endometriosis (mean +/- SEM, 91.1 +/- 6.5 pg/mL) than in control women (59.4 +/- 2.0 pg/mL). Interestingly, women with superficial (100.0 +/- 10.2 pg/mL) and deep peritoneal implants (94.0 +/- 10.8 pg/mL) had significantly higher PF IL-18 levels than did women with endometriomas (57.8 +/- 1.8 pg/mL). Similarly, women with stage I-II endometriosis (97.3 +/- 8.0 pg/mL), but not women with stage III-IV endometriosis (74.9 +/- 9.9 pg/mL), had significantly higher PF IL-18 levels than did control women. Peritoneal fluid IL 18 levels were significantly higher in the luteal phase than in the follicular phase but did not discriminate between women with pelvic pain or infertility. CONCLUSION(S): Peritoneal fluid IL-18 is elevated in women with peritoneal, minimal- to mild-stage endometriosis. PMID- 14556809 TI - Relationship between the outcomes of assisted reproductive techniques and sperm DNA fragmentation as measured by the sperm chromatin structure assay. AB - OBJECTIVE: To investigate how moderate and/or high levels of DNA fragmentation (DFI), as measured by the sperm chromatin structure assay (SCSA), affect either IVF or IVF with intracytoplasmic sperm injection (ICSI) fertilization, cleavage, blastulation, implantation, and pregnancy. DESIGN: Retrospective clinical study. SETTING: Academic human reproduction laboratory. PATIENT(S): Eighty-nine couples undergoing IVF with conventional fertilization or ICSI. INTERVENTION(S): Sperm chromatin structure assay testing (SCSA) of semen aliquot taken from ejaculate used for assisted reproductive technology (ART). MAIN OUTCOME MEASURE(S): Related DFI to conventional semen parameters and cycle-specific outcomes after ART. RESULT(S): No patients achieved clinical pregnancy if SCSA values exceeded the DFI (27%, P<.01), moderate DFI (15%, P<.01), or high DFI (15%, P<.05) thresholds. Dividing the DFI sperm population into moderate-fragmentation and high fragmentation categories did not improve the prognostic value of the SCSA. No coefficient of determination (r(2)) between SCSA parameters and conventional parameters exceeded 0.29. CONCLUSION(S): Sperm chromatin structure assay identified thresholds for negative pregnancy outcome after ART not identified using conventional semen parameters. This is the first study analyzing the clinical value of sperm DFI to [1] include a large number of ART patients (n = 89), [2] perform SCSA analysis on a semen aliquot from the ejaculate used for ART, and [3] examine how the extent (moderate and high DFI) of DFI influenced ART outcomes. PMID- 14556810 TI - Partners of men with Klinefelter syndrome can benefit from assisted reproductive technologies. AB - OBJECTIVE: To report the birth of a healthy female infant from a father with nonmosaic Klinefelter syndrome (KS) and document the experience of men with KS undergoing assisted conception. DESIGN: Retrospective. SETTING: Private IVF center. PATIENT(S): Twelve couples with male factor infertility due to Klinefelter syndrome undergoing assisted reproduction treatment. INTERVENTION(S): Controlled ovarian hyperstimulation, testicular sperm extraction, intracytoplasmic sperm injection (ICSI), round spermatid injection (ROSI), and preimplantation genetic diagnosis. MAIN OUTCOME MEASURE(S): Testicular sperm retrieval rate, fertilization rate, and pregnancy outcome. RESULT(S): There was a sufficient amount of motile sperm for injection into mature oocytes in 6 of the 11 testicular biopsies (54.5%). Fertilization rates for ICSI and ROSI cases were 54.2% and 41.6%, respectively. The pregnancy rate per ET was 27.2%. None of the ROSI cases resulted in pregnancy. Two patients had spontaneous abortions at 8 and 18 weeks of gestation, respectively. Only one patient delivered a healthy female baby after 36 weeks of an uneventful pregnancy. CONCLUSION(S): Men with KS can benefit from assisted reproductive technologies, and the testicular sperm retrieval rate among them is promising. Although sex chromosome aberrations among the embryos from men with KS are not common, couples can be offered preimplantation genetic diagnosis before ET. PMID- 14556811 TI - Scanning of Y-chromosome azoospermia factors loci using real-time polymerase chain reaction and melting curve analysis. AB - OBJECTIVE: To develop a novel method to scan AZF loci looking for microdeletions. DESIGN: Molecular method development. SETTING: Men undergoing reproductive techniques in a private fertility unit. Molecular methods were performed in a private center for biomedical research. PATIENT(S): : Fifty-eight men divided in two groups depending on seminal analyses. A group of 19 women were also included as positive controls (absence of amplification). INTERVENTION(S): Peripheral blood extraction and DNA purification. MAIN OUTCOME MEASURE(S): Our method is based on real-time polymerase chain reaction (PCR) and melting curve analysis. We performed the screening of 16 selected sequence tagged sites (STS) within AZF loci, and we also calculated the mean, range, and standard deviation for melting temperature patterns and the crossing points values for each STS tested. RESULT(S): We detected one azoospermic patient with several STS deleted within the AZFc region. No deletions were detected in a group of 13 healthy men, and no amplification for any of the STS tested were observed in the positive control group (19 healthy women). CONCLUSION(S): We have developed a novel method based on real-time PCR and melting curve analysis to scan AZF loci looking for microdeletions This method is fast and reliable and permits the scanning of DNA from one patient per hour, minimizing the risk of cross contamination, and false positive and false-negative results. PMID- 14556812 TI - Effectiveness of combined tamoxifen citrate and testosterone undecanoate treatment in men with idiopathic oligozoospermia. AB - OBJECTIVE: To assess the effect of treatment with a combination of the antiestrogen tamoxifen citrate and the androgen testosterone undecanoate on sperm variables and pregnancy incidence in men with idiopathic oligozoospermia and couple subfertility. DESIGN: Prospective, randomized, placebo-controlled trial. SETTING: Clinical research in a tertiary care academic hospital. PATIENT(S): Two hundred twelve men with idiopathic oligozoospermia and 82 normozoospermic men with female factor subfertility. INTERVENTION(S): Oligozoospermic patients were randomly assigned to two treatment groups with tamoxifen citrate, 20 mg/d, and testosterone undecanoate, 120 mg/d (n = 106) or placebo treatment (n = 106) for 6 months. Normozoospermic men were followed for the same period. Couple counseling was part of the intervention in all groups. MAIN OUTCOME MEASURE(S): Pregnancy incidence and sperm characteristics after 3 and 6 months on medication and 3 months after the end of the trial. RESULT(S): In the active treatment group, total sperm count (median [25th, 75th percentile], 27.1 x 10(6) cells/mL [9.4, 54.0 x 10(6) cells/mL] at baseline and 61.5 x 10(6) cells/mL [28.2, 119.6 x 10(6) cells/mL] at 6 months), progressive motility (mean [+/-SD], 29.7% +/- 12.0% at baseline and 41.6% +/- 13.1% at 6 months), and normal morphology (mean, 41.2% +/- 14.0% at baseline and 56.6% +/- 11.5% at 6 months) were noted. No marked changes were observed in placebo recipients or normozoospermic men. The incidence of spontaneous pregnancy was 33.9% in the active treatment group and 10.3% in the placebo group (36 vs. 11 pregnancies), with a relative risk of 3.195 (95% CI, 2.615 to 3.765). CONCLUSION(S): Treatment with tamoxifen citrate and testosterone undecanoate improved sperm variables and led to a higher incidence of pregnancy in couples with subfertility related to idiopathic oligozoospermia. PMID- 14556813 TI - Ureaplasma parvum and Ureaplasma urealyticum are detected in semen after washing before assisted reproductive technology procedures. AB - OBJECTIVE: To investigate the prevalence of ureaplasmas in semen and washed semen and to explore their effect on semen andrology variables. DESIGN: Prospective study. SETTING: In vitro fertilization (IVF) unit of a private hospital. PATIENT(S): Three hundred forty-three men participating in an assisted reproductive technology (ART) treatment cycle. MAIN OUTCOME MEASURE(S): The prevalence of ureaplasmas in semen and washed semen tested by culture, polymerase chain reaction assays, and indirect immunofluorescent antibody assays. RESULT(S): Ureaplasmas were detected in 73 of 343 (22%) semen samples and 29 of 343 (8.5%) washed semen samples. Ureaplasmas adherent to the surface of spermatozoa were demonstrated by indirect immunofluorescent antibody testing. Ureplasma parvum serovar 6 (36.6%) and U. urealyticum (30%) were the most prevalent isolates in washed semen. A comparison of the semen andrology variables of washed semen ureaplasma positive and negative groups demonstrated a lower proportion of nonmotile sperm in men ureaplasma positive for washed semen. CONCLUSION(S): Ureaplasmas are not always removed from semen by a standard ART washing procedure and can remain adherent to the surface of spermatozoa. PMID- 14556815 TI - Pregnancy outcomes using donor sperm insemination after failed in vitro fertilization with intracytoplasmic sperm injection cycles in couples with complex infertility disorders. AB - OBJECTIVE: To evaluate alternatives for couples with severe male factor infertility who fail to conceive with IVF-intracytoplasmic sperm injection (ICSI). DESIGN: Outcomes of couples using artificial insemination with donor sperm (AID) after failed IVF-ICSI, assessing multiple risk factors affecting prognosis. SETTING: University infertility service. PATIENT(S): Nineteen patients with complex infertility disorders who failed IVF-ICSI and subsequently used AID (1 to 7 cycles). INTERVENTION(S): Artificial insemination with donor sperm was performed 36 hours after detection of an LH surge or hCG injection. MAIN OUTCOME MEASURE(S): Pregnancy outcomes were determined. RESULT(S): Seventeen pregnancies occurred in 16 women associated with AID for a pregnancy rate per cycle of 27.9% within a mean of 3.2 +/- 18 cycles. Live birth rate per cycle was 24.6%. CONCLUSION(S): A high pregnancy rate was achieved with AID in women who failed IVF-ICSI. Given the low cost and effectiveness of AID in this series, consideration of AID is a reasonable and effective option even in couples with poor prognosis who fail to conceive with IVF-ICSI. PMID- 14556814 TI - Technological assessment of intracytoplasmic sperm injection: an analysis of the value context. AB - Since the introduction of intracytoplasmic sperm injection (ICSI) in 1992, the new technology has raised a series of basic moral issues. In this article, I present a brief review of this moral debate. I argue that even if we solve the moral problems of the debate, there is a residue of value issues. These value issues are crucial for the understanding of the dynamics of the health policy and the legislative process concerning ICSI. That is, values analyses are crucial to make viable moral judgments about new technologies such as ICSI. PMID- 14556816 TI - Modulation of human sperm function by peritoneal fluid. AB - OBJECTIVE: To examine the effect of peritoneal fluid on various parameters of sperm function in vitro. DESIGN: Prospective study. SETTING: Basic research laboratory. PATIENT(S): Semen samples were obtained from normozoospermic volunteers (n = 43). Peritoneal fluids were aspirated laparoscopically from women with unexplained infertility (n = 14). Follicular fluid and oocytes were collected from patients undergoing IVF-ET. INTERVENTION(S): Sperm incubated under capacitating conditions were exposed to peritoneal fluid, and functional variables were evaluated in vitro. MAIN OUTCOME MEASURE(S): Sperm viability and motility, follicular fluid and calcium ionophore-induced acrosome reactions, protein tyrosine phosphorylation, expression of D-mannose binding sites, and ability of sperm to interact with zona pellucida. RESULT(S): Exposure of sperm to peritoneal fluid for up to 6 hours did not affect sperm viability or motility. Unlike follicular fluid, peritoneal fluid did not induce the acrosome reaction. Moreover, incubation of sperm with > or =20% v/v peritoneal fluid for 1 hour prevented the follicular fluid and the ionophore-induced acrosome reaction. Although treatment with peritoneal fluid allowed protein tyrosine phosphorylation during capacitation, it resulted in a significant decrease in the expression of D mannose binding sites and sperm-zona pellucida binding. CONCLUSION(S): Peritoneal fluid maintains sperm survival and decreases sperm ability to respond to inducers of the acrosome reaction and bind to the zona pellucida in vitro, indicating that this fluid might modulate sperm function in vivo. PMID- 14556817 TI - Quantitation by image analysis of global DNA methylation in human spermatozoa and its prognostic value in in vitro fertilization: a preliminary study. AB - OBJECTIVE: To determine the relationship between sperm DNA methylation level and sperm characteristics and pregnancy rates. DESIGN: Prospective study. Quantitation by image analysis of DNA methylation in sperm nucleus. SETTING: Department of Reproduction Biology, Edouard Herriot Hospital, Lyon, France. PATIENT(S): Infertile couples undergoing IVF-ET. INTERVENTION(S): The immunostaining of 5 methyl-cytosine was performed on the spare sperm suspension that was used for an assisted reproduction technology procedure. MAIN OUTCOME MEASURE(S): Sperm characteristics according to World Health Organization criteria, sperm motility parameters with computer-assisted semen analysis, sperm DNA methylation level, and heterogeneity index (HI). RESULT(S): Sperm DNA methylation level and HI are correlated with sperm DNA characteristics. HI is negatively correlated with fertilization rate; sperm DNA methylation level is correlated with pregnancy rate. CONCLUSION(S): The DNA methylation level in human spermatozoa could be a new approach to evaluating the ability of spermatozoa to fertilize and lead to normal embryo development. PMID- 14556818 TI - Selective ovary resistance to insulin signaling in women with polycystic ovary syndrome. AB - OBJECTIVE: Insulin resistance is a common feature of both polycystic ovary syndrome (PCOS) and non-insulin-dependent diabetes mellitus (NIDDM); however, the persistent reproductive disturbances appear to be limited to the former, suggesting that insulin resistance in the ovary itself may confer this susceptibility. DESIGN: Prospective study. SETTING: University-affiliated department. PATIENT(S): Forty-four women undergoing IVF treatment, of whom 11 had polycystic ovaries and 33 had normal ovulation (NO). INTERVENTION(S): The various effects and signaling of insulin and insulin-like growth factor-1 (IGF-1) were examined in cultured ovarian granulosa cells treated with troglitazone (1 microg/mL) or with vehicle by reverse transcription-polymerase chain reaction, western blot, and in vitro functional analyses. MAIN OUTCOME MEASURE(S): Glycogen and DNA syntheses, mRNA and protein expression, and cellular localization of insulin/IGF-1 receptors and insulin receptor substrates (IRSs). RESULT(S): There were significant decreases in insulin-stimulated glucose incorporation into glycogen in PCOS cells, which is a metabolic action of insulin. However, IGF-1 stimulation was found to be greater in PCOS cells at all experimental concentrations with respect to thymidine incorporation compared with NO cells, which is a mitogenic action. Troglitazone increased the insulin-induced glycogen synthesis but reduced the IGF-1-augmented responses of DNA synthesis in PCOS cells to the range within those of NO granulosa cells. We then found that troglitazone treatment reversed the expression imbalance between IRS-1 and IRS-2 in PCOS cells. CONCLUSION(S): There is a selective defect in insulin actions in PCOS granulosa cells, which suggests ovarian insulin resistance, and this metabolic phenotype is associated with an enhanced IGF-1 mitogenic potential. Troglitazone could divergently alter expression of various IRS molecules and insulin actions and could be used as an ovarian insulin sensitizer and mitogen/steroidogenic inhibitor in PCOS. PMID- 14556819 TI - Ultrasound diagnosis of polycystic ovaries in women who have no symptoms of polycystic ovary syndrome is not associated with subfecundity or subfertility. AB - OBJECTIVE: To evaluate the effect on fertility of the appearance of polycystic ovaries in women who have no symptoms of polycystic ovary syndrome. DESIGN: Case control study. SETTING: Teaching hospitals in Hull, United Kingdom. PATIENT(S): Women with the appearance of polycystic ovaries on ultrasound and women with normal ovaries. INTERVENTION(S): A questionnaire about previous subfertility, pregnancies, menstrual pattern, features of polycystic ovary syndrome, gynecological history, and individual lifestyle factors. MAIN OUTCOME MEASURE(S): Time to pregnancy (TTP) and relative risk (RR) of subfertility in symptomatic and asymptomatic subgroups of both groups. RESULT(S): Women with PCOs took longer TTP and were significantly less fertile if they were obese (RR = 2.6), had menstrual disturbances (RR = 4.6), hirsutism (RR = 2.5), and/or acne (RR = 2.7). Further reductions in fecundity occurred with an increasing number of symptoms (threefold, sevenfold, and 10-fold longer TTP with two, three, and four symptoms, respectively). The TTP of women with no symptoms was not significantly longer and they were not more likely to be subfertile than women with normal ovaries. These symptoms were not associated with significantly reduced fecundity in women with normal ovaries. CONCLUSION(S): The appearance of polycystic ovaries has been shown to have no significant impact on fertility in women with no symptoms. Appearances alone do not reflect the pathological features of polycystic ovary syndrome, and additional diagnostic criteria should be considered. Obesity, menstrual disturbances, and/or hyperandrogenism are factors associated with subfertility in women with polycystic ovaries. PMID- 14556820 TI - Recovery of ovarian activity in women with functional hypothalamic amenorrhea who were treated with cognitive behavior therapy. AB - OBJECTIVE: To determine whether cognitive behavior therapy (CBT) targeted to problematic attitudes common among women with functional hypothalamic amenorrhea would restore ovarian function. DESIGN: Randomized, prospective, controlled intervention. SETTING: Clinical research center in an academic medical institution. PATIENT(S): Sixteen women participated who had functional hypothalamic amenorrhea; were of normal body weight; and did not report psychiatric conditions, eating disorders, or excessive exercise. INTERVENTION(S): Subjects were randomized to CBT or observation for 20 weeks. MAIN OUTCOME MEASURE(S): Serum levels of E(2) and P and vaginal bleeding were monitored. RESULT(S): Of eight women treated with CBT, six resumed ovulating, one had partial recovery of ovarian function without evidence of ovulation, and one did not display return of ovarian function. Of those randomized to observation, one resumed ovulating, one had partial return of ovarian function, and six did not recover. Thus, CBT resulted in a higher rate of ovarian activity (87.5%) than did observation (25.0%), chi(2) = 7.14. CONCLUSION(S): A cognitive behavioral intervention designed to minimize problematic attitudes linked to hypothalamic allostasis was more likely to result in resumption of ovarian activity than observation. The prompt ovarian response to CBT suggests that a tailored behavioral intervention offers an efficacious treatment option that also avoids the pitfalls of pharmacological modalities. PMID- 14556821 TI - Treatment of functional hypothalamic amenorrhea with hypnotherapy. AB - OBJECTIVE: To determine the effects of hypnotherapy on resumption of menstruation in patients with functional hypothalamic amenorrhea (FHA). DESIGN: Uncontrolled clinical study. SETTING: Academic clinical care center. PATIENT(S): Twelve consecutive women with FHA were selected. INTERVENTION(S): A single 45- to 70 minute session of hypnotherapy was administered, and patients were observed for 12 weeks. MAIN OUTCOME MEASURE(S): Patients were asked whether or not menstruation resumed and whether or not well-being and self-confidence changed. RESULT(S): Within 12 weeks, 9 out of 12 patients (75%) resumed menstruation. All of the patients, including those who did not menstruate, reported several beneficial side effects such as increased general well-being and increased self confidence. CONCLUSION(S): Hypnotherapy could be an efficacious and time-saving treatment option that also avoids the pitfalls of pharmacological modalities for women with FHA. PMID- 14556822 TI - Follicle-stimulating hormone receptor polymorphisms in women with normogonadotropic anovulatory infertility. AB - OBJECTIVE: To assess the incidence of different FSH receptor genotypes in normogonadotropic anovulatory infertile women (World Health Organization class II) and normo-ovulatory controls and to correlate these genotypes with baseline characteristics and ovarian responsiveness during ovulation induction. DESIGN: Cross-sectional study. SETTING: University hospital. PATIENT(S): Thirty normo ovulatory controls and 148 normogonadotropic anovulatory infertile women. INTERVENTION(S): All participants underwent a standardized evaluation that included cycle history, body mass index measurement, and transvaginal ultrasonography of ovaries. Fasting blood samples were obtained for endocrine evaluation. Ovarian responsiveness to FSH in normogonadotropic anovulatory infertile women was assessed during ovulation induction, and DNA was analyzed to determine the FSH receptor genotype. MAIN OUTCOME MEASURE(S): Prevalence of FSH receptor polymorphisms, baseline serum FSH levels, amount of FSH administered, duration of stimulation, and ovarian response dose. RESULT(S): The Thr/Thr 307 genotype was significantly less prevalent (52% vs. 23%) and the Ser/Ser 680 polymorphism was significantly more prevalent (40% vs. 16%) in patients compared with controls. Normogonadotropic anovulatory infertile women with the Ser/Ser 680 polymorphism presented with higher median FSH serum levels (5.2 IU/L [range, 2.4 9.7 IU/L]) than did those with the Asn/Asn 680 (4.6 IU/L [range, 1.4-5.8 IU/L) and Asn/Ser 680 (4.5 IU/L [range, 1.8-9.7 IU/L) variants. However, ovarian responsiveness to FSH was similar among anovulatory women with the various polymorphisms. CONCLUSION(S): Normogonadotropic anovulatory infertile patients have a different FSH receptor genotype than do normo-ovulatory controls. Although this characteristic is associated with increased baseline FSH serum levels, altered ovarian sensitivity to exogenous FSH during ovulation induction could not be established. PMID- 14556823 TI - ATP-induced apoptosis of human granulosa luteal cells cultured in vitro. AB - OBJECTIVE: To investigate the role of extracellular adenosine triphosphatase (ATP) as an inducer of apoptotic cell death in human granulosa cells and to elucidate its underlying mechanism. DESIGN: Prospective study. SETTING: Gynecologic clinic and human reproduction research laboratory. PATIENT(S): Twenty five patients undergoing IVF or IVF-ET. INTERVENTION(S): ATP treatment of granulosa luteal cells subjected to primary culture. MAIN OUTCOME MEASURE(S): Apoptosis was assessed by the annexin V binding assay and the terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling assay. The receptor type that binds ATP, thus mediating ATP-induced apoptosis, was determined by using the calcium imaging and patch-clamp techniques. Mitochondrial depolarization was assessed by staining with the membrane potential-sensitive dye 5,5',6,6'-tetrachloro-1,1',3,3'- tetraethylbenzimidazolyl carbocyanide iodide (JC 1). RESULT(S): ATP elevated [Ca(2+)](i) by mobilizing intracellularly stored Ca(2+). An ionic membrane current evoked by ATP was mediated by the Ca(2+) activated K(+) channel. ATP induced a mitochondrial depolarization with a concomitant increase in cellular apoptosis. Treatment with hCG eliminated both ATP-induced mitochondrial depolarization and apoptosis. CONCLUSION(S): Extracellular ATP recognized by P(2Y) type purinoceptor triggers apoptosis in human granulosa luteal cells, and the downstream apoptotic cascade may act at least in part through mitochondria. The antiapoptotic effect of hCG has possible clinical implications in the treatment of disorders involving granulosa cell degeneration (such as follicular atresia). PMID- 14556824 TI - Karyotyping of human metaphase II oocytes by multifluor fluorescence in situ hybridization. AB - OBJECTIVE: To quantify aneuploidy in inseminated, injected, and noninjected oocytes from infertility patients using Multifluor fluorescence in situ hybridization (M-FISH). DESIGN: Prospective study. SETTING: Reproductive biology group, academic unit of pediatrics, obstetrics, and gynecology. PATIENT(S): Forty eight patients undergoing ovarian stimulation and either intracytoplasmic sperm injection (ICSI) or conventional in vitro fertilization (IVF). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): M-FISH karyotyping of 67 metaphase II oocytes, including noninjected in vitro matured oocytes, and injected inseminated-failed fertilized oocytes. RESULT(S): Thirty-nine percent of oocytes were aneuploid, with nondisjunction of chromosomes in 34% of oocytes and predivision of chromatids in 10%. There was no difference in aneuploidy rates between ICSI noninjected in vitro matured oocytes and injected, failed fertilized oocytes. Chromosomes most frequently involved in aneuploidy were 15, 18, 19, 22, and X. In seven injected ICSI MII oocytes, the prematurely condensed sperm chromatin was karyotyped by M-FISH. CONCLUSION(S): M-FISH was used to diagnose aneuploidy at maternal meiosis I in 39% of oocytes, and M-FISH karyotyping of sperm was demonstrated. PMID- 14556825 TI - Increased endogenous level of melatonin in preovulatory human follicles does not directly influence progesterone production. AB - OBJECTIVE: To study the role of follicular melatonin on steroid production by human preovulatory follicles. DESIGN: In vivo comparative and in vitro culture studies. SETTING: University hospital. PATIENT(S): Thirty-six patients with tubal and/or male, but not endocrinological, infertility factors. INTERVENTION(S): Follicular fluid collection during IVF. In vitro granulosa cell culture from luteinizing or growing Graafian follicles. MAIN OUTCOME MEASURE(S): Follicular melatonin, P, T, and E(2) concentrations. In vitro P production by granulosa cells. RESULT(S): There was a positive correlation between follicular melatonin and P concentrations. Melatonin did not stimulate in vitro P production by granulosa cells from luteinizing or growing follicles. Melatonin, P, and E(2) concentrations were significantly higher, but T concentrations were lower, in large follicles than in small follicles. CONCLUSION(S): Preovulatory follicles contain a high amount of melatonin compared with that in small immature follicles; melatonin may play an important, but indirect, role in P production by human granulosa cells. PMID- 14556826 TI - Short coincubation of gametes in in vitro fertilization improves implantation and pregnancy rates: a prospective, randomized, controlled study. AB - OBJECTIVE: To examine the benefits of short coincubation of gametes compared to prolonged incubation in in vitro fertilization (IVF). DESIGN: Prospective randomized controlled study. SETTING: Hospital-based fertility center. PATIENT(S): One hundred thirty patients (group 1) and 129 patients (group 2). INTERVENTION(S): Oocytes from group 1 were exposed to spermatozoa for 2 hours, and oocytes from group 2 were exposed to spermatozoa for 20 hours. MAIN OUTCOME MEASURE(S): Fertilization and cleavage rates, embryo quality, and pregnancy and implantation rates were evaluated. Estradiol (E(2)) and progesterone (P(4)) levels were measured in the wells of culture dishes after 2-hour exposure of oocytes/zygotes to spermatozoa in group 1 and after 20-hour exposure in both the groups. RESULT(S): There was no difference between the two groups in the fertilization rate and the number of embryos obtained. However, the proportion of grade 1 embryos was significantly higher among group 1 compared to group 2 patients. Clinical pregnancy and implantation rates were significantly higher among group 1 compared to group 2. The significantly higher levels of E(2) and P(4) in the 20-hour cultures compared with the 2-hour cultures may have been detrimental to embryo quality, pregnancy, and implantation rates. CONCLUSION(S): Coincubation of gametes for 2 hours with standard or high insemination concentrations of spermatozoa significantly improved embryo quality and the pregnancy and implantation rates compared with overnight incubation of gametes. PMID- 14556827 TI - Values of fasting glucose levels, glucose tolerance tests, and glucose-insulin ratios as predictors of glucose tolerance. AB - OBJECTIVE: To examine the ability to use parameters obtainable from an oral glucose tolerance test to predict insulin action as determined under hyperinsulinemic, hyperglycemic conditions. DESIGN: Prospective clinical investigation. SETTING: University medical center clinical research unit. PATIENT(S): Healthy male volunteers. INTERVENTION(S): Oral glucose tolerance test and hyperglycemic (+125 mg/dL) clamp studies. MAIN OUTCOME MEASURE(S): Glucose and insulin (I) levels, rate of glucose uptake (M) under hyperglycemic conditions, and M/I ratios. RESULT(S): Among individuals with normal glucose tolerance, as assessed by an oral glucose tolerance test, the fasting insulin level is the glucose tolerance test parameter that correlates best with insulin action during a hyperglycemic clamp. CONCLUSION(S): Measurement of fasting serum insulin levels in conjunction with an oral glucose tolerance test improves the ability to assess insulin action. Such combinations may improve the ability to diagnose insulin-resistant states. PMID- 14556828 TI - Angiopoietin-2 polymorphism in women with idiopathic recurrent miscarriage. AB - OBJECTIVE: To investigate the relationship between idiopathic recurrent miscarriage and a polymorphism of the gene encoding for angiopoietin-2 (ANGPT2), an autochthonous modulator of angiogenesis during pregnancy. DESIGN: Prospective case control study. SETTING: Academic research institution. PATIENT(S): One hundred thirty-one women with a history of three or more consecutive pregnancy losses before 20 weeks' gestation, and 125 healthy, postmenopausal controls with at least two live births and no history of pregnancy loss. INTERVENTION(S): Peripheral venous puncture. MAIN OUTCOME MEASURE(S): Polymerase chain reaction and restriction fragment length polymorphism analysis were performed to identify the different ANGPT2 alleles. RESULT(S): No association between mutant (mt) allele and the occurrence of idiopathic recurrent miscarriage was found. Between women with primary and secondary idiopathic recurrent miscarriage, no statistically significant differences with respect to allele frequencies were observed. CONCLUSION(S): This is the first report on the ANGPT2 gene polymorphism in women with idiopathic recurrent miscarriage, demonstrating that the investigated polymorphism is not associated with idiopathic recurrent miscarriage in a white population. PMID- 14556829 TI - Monitoring of menstrual cycles, ovulation, and adrenal suppression by saliva sampling in female patients with 21-hydroxylase deficiency. AB - OBJECTIVE: To investigate the correlation between menstrual cycles, ovulation, and adrenal suppression in congenital adrenal hyperplasia. DESIGN: Prospective observational study. SETTING: An academic outpatient clinic. PATIENT(S): Five females with salt-wasting 21-hydroxylase deficiency, aged 15.5 to 22.9 years; one had amenorrhea, one had irregular bleeding, and three had regular bleeding. INTERVENTION(S): Daily morning saliva sampling for 40 to 280 days. MAIN OUTCOME MEASURE(S): Salivary levels of progesterone (P), 17-hydroxyprogesterone (17-OHP), and androstenedione. RESULT(S): In the amenorrheic patient, the elevated P and 17 OHP levels decreased when the glucocorticoid dose was increased, and subsequently menarche occurred. The androstenedione levels were normal. The correlations between P and 17-OHP levels before and after menarche suggest that adrenal progesterone had prevented menarche. The patient with irregular bleeding showed slightly elevated androstenedione levels and increased levels of 17-OHP and P in an irregular pattern, without correlation in time with vaginal bleeding. Three patients with regular cycles showed a biphasic pattern of P levels, indicating ovulation. CONCLUSION(S): These longitudinal data support the hypothesis that menstrual cycling in females with 21-hydroxylase deficiency can be prevented or disturbed by elevated progesterone levels of adrenal origin, in the absence of androgen excess. Increasing glucocorticoid dose could suppress adrenal progesterone production, resulting in menarche. PMID- 14556830 TI - A meta-analysis of ultrasound-guided versus clinical touch embryo transfer. AB - OBJECTIVE: To determine the relative efficacy of ultrasound-guided embryo transfer and embryo transfer by clinical touch alone. DESIGN: Systematic review and meta-analysis of randomized, controlled trials comparing ultrasound-guided embryo transfer with embryo transfer by clinical touch alone. SETTING: Infertility centers providing treatment with in vitro fertilization/embryo transfer. PATIENT(S): Women undergoing embryo transfer. INTERVENTION(S): Embryo transfer with or without transabdominal ultrasound guidance. MAIN OUTCOME MEASURE(S): Clinical pregnancy rate and embryo implantation rate. RESULT(S): A total of eight prospective controlled trials were identified. Of these studies, four were nonrandomized or quasi-randomized and four were genuinely randomized. Meta-analysis demonstrated a significantly increased chance of clinical pregnancy following ultrasound-guided embryo transfer in all studies and in the genuinely randomized subgroup. The embryo implantation rate was also significantly increased following ultrasound-guided embryo transfer. CONCLUSION(S): Ultrasound guided embryo transfer significantly increases the chance of clinical pregnancy and significantly increases the embryo implantation rate. PMID- 14556831 TI - Ultrasound-guided embryo transfer: a meta-analysis of randomized controlled trials. AB - OBJECTIVE: To conduct a meta-analysis of randomized controlled studies to evaluate abdominal ultrasound-guided embryo transfer compared to the clinical touch method. DESIGN: Meta-analysis. SETTING: Four studies performed in academic research centers in Barcelona, Vizcaya, Hong Kong, and Madrid. PATIENT(S): A total of 2,051 patients: 1,024 received ultrasound-guided embryo transfers and 1,027 received clinical touch method transfers. INTERVENTION(S): Fourteen studies were retrieved from the literature, but only four of them fitted our selection criteria. The meta-analysis was conducted using the RevMan software with the Peto modified Mantel-Haenszel method and the fixed effect model. MAIN OUTCOME MEASURE(S): The primary outcome measures were the clinical pregnancy and ongoing pregnancy rates. The secondary outcome measures were the incidences of ectopic pregnancies, miscarriage rate, multiple pregnancies, and difficult transfers. RESULT(S): Compared to the clinical touch method, abdominal ultrasound-guided transfer significantly increased the clinical pregnancy rate and the ongoing pregnancy rate. There was no effect on the incidence of ectopic pregnancy, multiple pregnancy, or miscarriage rate. CONCLSUION(S): Ultrasound-guided embryo transfer increases the clinical pregnancy and ongoing pregnancy rates significantly compared to the clinical touch method. PMID- 14556832 TI - Preimplantation genetic diagnosis for the Kell genotype. AB - OBJECTIVE: To use preimplantation genetic diagnosis (PGD) to achieve a Kell 1 (K1) allele-free pregnancy in couples at risk for producing a child with hemolytic disease of the newborn (HDN) caused by maternofetal incompatibility in sensitized mothers. DESIGN: DNA analysis of biopsied blastomeres from cleavage stage embryos in IVF-ET with the goal of identifying and transferring back to patients the K1 allele-free embryos. SETTING: IVF program at the Reproductive Genetics Institute, Chicago, Illinois, and IVF Michigan, Rochester Hills, Michigan. PATIENT(S): Two at-risk couples with a history of neonatal death caused by HDN due to K1/K2 genotype in a male partner. INTERVENTION(S): Biopsy of single blastomeres and testing for paternal K1 allele in each embryo after standard IVF. MAIN OUTCOME MEASURE(S): DNA analysis of blastomeres indicating whether corresponding embryos were K1 allele-free for the purpose of transferring only embryos without the K1 allele. RESULT(S): Of 36 embryos tested in five cycles from two couples, 18 were predicted to be K1 allele-free. Of these, 9 were transferred, resulting in a K1 allele-free twin pregnancy and the birth of two healthy children. CONCLUSION(S): PGD of the K1 genotype resulted in the birth of healthy twins confirmed to be free of the K1 allele. PGD in couples with a heterozygous K1/K2 male partner provides an option for avoiding HDN in sensitized mothers. PMID- 14556834 TI - Predictive value of urine human chorionic gonadotropin after assisted reproductive technology. PMID- 14556833 TI - Prevalence of the triple X syndrome in phenotypically normal women with premature ovarian failure and its association with autoimmune thyroid disorders. AB - OBJECTIVE: To determine the prevalence of triple X females among patients with premature ovarian failure and to describe the clinical features of the syndrome. DESIGN: Case report. SETTING: Tertiary care hospital. PATIENT(S): Fifty-two consecutive patients with secondary amenorrhea due to premature ovarian failure and no clinical stigmata of Turner's syndrome. MAIN OUTCOME MEASURE(S): Triple X syndrome and clinical features, as assessed by karyotype analysis using Giemsa trypsin banding of metaphase chromosomes. RESULT(S): Two of the 52 patients with premature ovarian failure had triple X syndrome. Both cases had associated autoimmune thyroid disorder. One of the women with triple X syndrome had two pregnancies that were complicated by premature birth, idiopathic thrombocytopenia, neonatal death, and occipital encephalocoele. CONCLUSION(S): Among patients with premature ovarian failure, 3.8% have triple X syndrome. The syndrome may be associated with autoimmune thyroid disorder and poor pregnancy outcome due to congenital malformation. PMID- 14556835 TI - Quantitative determination of sst2 and sst5 gene expression in uterine leiomyomata and the effect of treatment with somatostatin analogue. PMID- 14556836 TI - Effects of goserelin administration before myomectomy on plasma homocysteine levels in patients with symptomatic uterine leiomyomata. PMID- 14556837 TI - Autologous transplantation of cryopreserved ovary induces the generation of antiovary antibodies in sheep. PMID- 14556838 TI - High CD44 content in ovarian endometriotic cysts. PMID- 14556839 TI - A critical issue in the summary measures for ART. What is the experimental unit- the cycle or the patient? PMID- 14556841 TI - Polycystic ovarian disease and serum leptin levels? PMID- 14556843 TI - Optimal day for embryo transfer? PMID- 14556845 TI - Diagnosis of endometriosis: utility of MRI? PMID- 14556848 TI - Metabolism of dietary procyanidins in rats. AB - Procyanidins are major dietary polyphenols made of elementary flavan-3-ol (epi)catechin units. They have antioxidant properties and may contribute to health benefits in humans, but little is known about their metabolic fate. We compared here the metabolism of procyanidin dimer B3, trimer C2, and polymer isolated from willow tree catkins to that of catechin monomer in rats. These compounds were administered in the rat diet (0.1%, w/w) for 5 d and their metabolites estimated in 24 h urine. In rats fed procyanidins, neither parent compound nor catechin derivatives could be detected in contrast to animals fed catechin monomer, which excreted large amounts of catechin and its 3'-O methylated form (25.7 +/- 0.6%). On the other hand, 16 metabolites of microbial origin were detected and identified as phenylvaleric, phenylpropionic, phenylacetic, and benzoic acid derivatives. Their total yields significantly decreased from the catechin monomer (10.6 +/- 1.1%) to the procyanidin dimer (6.5 +/- 0.2%), trimer (0.7 +/- 0.1%), and polymer (0.5 +/- 0.1%). Therefore, the degree of procyanidin polymerization has a major impact on their fate in the body characterized by a poor absorption through the gut barrier and a limited metabolism by the intestinal microflora as compared to catechin. This will have to be considered to explain the health effects of procyanidins. The contribution of their microbial metabolites should also be further investigated. PMID- 14556849 TI - Vanadium-induced STAT-1 activation in lung myofibroblasts requires H2O2 and P38 MAP kinase. AB - Vanadium compounds present in air pollution particulate matter activate signal transduction pathways in pulmonary cell types leading to pathological outcomes including aberrant cell proliferation, apoptosis, and cytokine expression. Vanadium has been proposed to activate transcription factors via the generation of hydrogen peroxide (H2O2). We investigated the mechanisms through which vanadium pentoxide (V2O5), the major form of vanadium released from the industrial burning of fuel oil, activated the signal transducer and activator of transcription (STAT)-1. V2O5-induced STAT-1 activation was blocked by catalase and N-acetyl-L-cysteine (NAC), suggesting vanadium-induced generation of H2O2. Surprisingly, however, V2O5 did not increase H2O2 levels released by rat lung myofibroblasts into cell culture supernatants. Instead, these quiescent myofibroblasts spontaneously released micromolar concentrations of H2O2, and the addition of V2O5 reduced H2O2 levels in cell culture supernatants within minutes. V2O5 suppressed H2O2 for as long as 24 h. Differences in the temporal activation of STAT-1 and p38 MAPK were observed following V2O5 or H2O2 treatment, and STAT-1 activation by V2O5 or H2O2 was attenuated by an inhibitor of the EGF receptor tyrosine kinase (AG1478) or p38 MAPK (SB203580). The phosphorylation of p38 MAPK by V2O5 was inhibited by NAC and catalase, yet the EGF receptor inhibitor AG1478 had no effect on V2O5-induced p38 MAPK activation. Collectively, our findings support the novel hypothesis that H2O2 spontaneously generated by myofibroblasts fuels vanadium-induced activation of STAT-1. Moreover, p38 MAPK and EGF receptor activation are required for V2O5-induced STAT-1 activation. PMID- 14556850 TI - Scavenging system efficiency is crucial for cell resistance to ROS-mediated methylglyoxal injury. AB - Methylglyoxal is a reactive dicarbonyl compound endogenously produced mainly from glycolytic intermediates. Recent research indicates that methylglyoxal is a potent growth inhibitor and genotoxic agent. The antiproliferative activity of methylglyoxal has been investigated for pharmacological application in cancer chemotherapy. However, various cells are not equally sensitive to methylglyoxal toxicity. Therefore, it would be important to establish the cellular factors responsible for the different cell-type specific response to methylglyoxal injury, in order to avoid the risk of failure of a therapy based on increasing the intracellular level of methylglyoxal. To this purpose, we comparatively evaluated the signaling transduction pathway elicited by methylglyoxal in human glioblastoma (ADF) and neuroblastoma (SH-SY 5Y) cells. Results show that methylglyoxal causes early and extensive reactive oxygen species generation in both cell lines. However, SH-SY 5Y cells show higher sensitivity to methylglyoxal challenge due to a defective antioxidant and detoxifying ability that, preventing these cells from an efficient scavenging action, elicits extensive caspase-9 dependent apoptosis. These data emphasize the pivotal role of antioxidant and detoxifying systems in determining the grade of sensitivity of cells to methylglyoxal. PMID- 14556851 TI - Heme oxygenase-2 knockout neurons are less vulnerable to hemoglobin toxicity. AB - When cortical neurons are exposed to hemoglobin, they undergo oxidative stress that ultimately results in iron-dependent cell death. Heme oxygenase (HO)-2 is constitutively expressed in neurons and catalyzes heme breakdown. Its role in the cellular response to hemoglobin is unclear. We tested the hypothesis that HO-2 attenuates hemoglobin neurotoxicity by comparing reactive oxygen species (ROS) formation and cell death in wild-type and HO-2 knockout cortical cultures. Consistent with prior observations, hemoglobin increased ROS generation, detected by fluorescence intensity after dihydrorhodamine 123 or dichlorofluorescin diacetate loading, in wild-type neurons. This fluorescence was significantly attenuated in cultures prepared from HO-2 knockout mice, and cell death as determined by propidium iodide staining was decreased. In other experiments, hemoglobin exposure was continued for 19 h; cell death as quantified by LDH release was decreased in knockout cultures, and was further diminished by treatment with the HO inhibitor tin protoporphyrin IX. In contrast, HO-2 knockout neurons were more vulnerable than wild-type neurons to inorganic iron. HO-1, ferritin, and superoxide dismutase expression in HO-2 -/- cultures did not differ significantly from that observed in HO-2 +/+ cultures; cellular glutathione levels were slightly higher in knockout cultures. These results suggest that heme breakdown by heme oxygenase accelerates the oxidative neurotoxicity of hemoglobin, and may contribute to neuronal injury after CNS hemorrhage. PMID- 14556852 TI - The effects of hydrogen peroxide promoted by homocysteine and inherited catalase deficiency on human hypocatalasemic patients. AB - Elevated plasma homocysteine can generate oxygen free radicals and hydrogen peroxide. The enzyme catalase is involved in the protection against hydrogen peroxide. We examined the effect of oxidative stress promoted by homocysteine on erythrocyte metabolism (blood hemoglobin, MCV, folate, B12, serum LDH, LDH isoenzymes, haptoglobin) in the oxidative stress sensitive Hungarian patients with inherited catalase deficiency. The plasma homocysteine (HPLC method, Bio Rad), folate, B12 (capture binding assay, Abbott), blood hemoglobin concentrations, blood catalase activity (spectrophotometric assay of hydrogen peroxide), and MCV values were determined in 7 hypocatalasemic families including hypocatalasemic (male:12, female:18) patients and their results were compared to those of the normocatalasemic (male:17 female: 12) family members. We found decreased (p <.036) folate (ng/ml) concentrations (male hypocatalasemic 5.44 +/- 2.81 vs. normocatalasemic 7.56 +/- 1.97, female 5.01 +/- 1.93 vs. 6.61 +/- 1.91), blood hemoglobin (p <.010, male:140.2 +/- 11.0 vs. 153.6 +/- 11.6 g/l, female: 128.4 +/- 10.9 vs. 139.6 +/- 9.2 g/l). Increased levels of MCV (p <.001) were detected in hypocatalasemic patients (male: 98.6 +/- 3.4 vs. 90.1 +/- 7.5 fl, female: 95.9 +/- 3.9 vs. 90.1 +/- 2.5 fl), plasma homocysteine (p <.049, male: 9.72 +/- 3.61 vs. 7.36 +/- 2.10 umol/l, female: 9.06 +/- 3.10 vs. 6.84 +/- 2.50 umol/l) and not significant (p >.401) plasma B12 (male: 336 +/- 108 vs. 307 +/- 76 pg/ml, female: 373 +/- 180 vs. 342 +/- 75 pg/ml). The serum markers of hemolysis (LDH, LDH isoenzymes, haptoglobin) did not show significant (p >.228) signs of oxidative erythrocyte damage. We report firstly on increased plasma homocysteine concentrations in inherited catalase deficiency. The increased plasma homocysteine and inherited catalase deficiency together could promote oxidative stress via hydrogen peroxide. The patients with inherited catalase deficiency are more sensitive to oxidative stress of hydrogen peroxide than the normocatalasemic family members. This oxidative stress might be responsible for the decreased concentration of the blood hemoglobin via the oxidation sensitive folate and may contribute to the early development of arteriosclerosis and diabetes in these patients. PMID- 14556853 TI - Two redox centers within Yap1 for H2O2 and thiol-reactive chemicals signaling. AB - The Yap1 transcription factor regulates yeast responses to H2O2 and to several unrelated chemicals and metals. Activation by H2O2 involves Yap1 Cys303-Cys598 intra-molecular disulfide bond formation directed by the H2O2 sensor Orp1/Gpx3. We show here that the electrophile N-ethylmaleimide activates Yap1 by covalent modification of Yap1 C-terminal Cys598, Cys620, and Cys629, in an Orp1 and Yap1 oxidation-independent way, thus establishing an alternate and distinct mode of Yap1 activation. We also show that menadione, a superoxide anion generator and a highly reactive electrophile, operates both modes of Yap1 activation. Further, the Yap1 C-terminal domain reactivity towards other electrophiles (4 hydroxynonenal, iodoacetamide) and metals (cadmium, selenium) suggests a common mechanism for sensing thiol reactive chemicals, involving thiol chemical modification. We propose that Yap1 has two distinct molecular redox centers, one triggered by ROS (hydroperoxides and the superoxide anion) and the other by chemicals with thiol reactivity (electrophiles and divalent heavy metals cations). These data indicate that yeast cells cannot sense these compounds through the same molecular devices, albeit they are all electrophilic. PMID- 14556854 TI - Impact of intraischemic temperature on oxidative stress during hepatic reperfusion. AB - This study was designed to investigate the influence of intraischemic liver temperature on oxidative stress during postischemic normothermic reperfusion. In C57BL/6 mice, partial hepatic ischemia was induced for 90 min and intraischemic organ temperature adjusted to 4 degrees C, 15 degrees C, 26 degrees C, 32 degrees C, and 37 degrees C. As detected by electron spin-resonance spectroscopy, plasma/blood concentrations of hydroxyl and ascorbyl radicals were significantly increased in all groups after ischemia/reperfusion independent of the intraischemic temperature. In tissue, however, postischemic lipid peroxidation was attenuated after organ cooling down to 32 degrees C-26 degrees C and not detectable after ischemia at 15 degrees C-4 degrees C. mRNA expression of superoxide dismutase-1 and heme oxygenase-1, measured during reperfusion, was significantly elevated in the group at 37 degrees C as compared to the hypothermic groups at 4 degrees C-32 degrees C. The reduction of radical generation was associated with a prevention of adenosine monophosphate hydrolysis during ischemia in the hypothermic groups. In conclusion, ischemia-reperfusion induced oxidative stress in the liver tissue is non-linearly-dependent on intraischemic temperature, whereas the plasma/blood concentration of radicals is not affected by organ cooling. Oxidative stress is reduced through mild hypothermia at 32 degrees C-26 degrees C and inhibited completely at 15 degrees C. Reduction of initial intracellular radical generation and prevention of secondary oxidant-induced tissue injury are possible mechanisms of this protection. PMID- 14556855 TI - Epicatechin and its methylated metabolite attenuate UVA-induced oxidative damage to human skin fibroblasts. AB - The ultraviolet A component of sunlight causes both acute and chronic damage to human skin. In this study the potential of epicatechin, an abundant dietary flavanol, and 3'-O-methyl epicatechin, one of its major in vivo metabolites, to protect against UVA-induced damage was examined using cultured human skin fibroblasts as an in vitro model. The results obtained clearly show that both epicatechin and its metabolite protect these fibroblasts against UVA damage and cell death. The hydrogen-donating antioxidant properties of these compounds are probably not the mediators of this protective response. The protection is a consequence of induction of resistance to UVA mediated by the compounds and involves newly synthesized proteins. The study provides clear evidence that this dietary flavanol has the potential to protect human skin against the deleterious effects of sunlight. PMID- 14556856 TI - Serum ferritin concentration is associated with plasma levels of cholesterol oxidation products in man. AB - Cholesterol oxidation products, oxysterols, are thought to play a part in the initiation and development of human atherosclerotic lesions. Excessive body iron has been suggested to promote atherosclerosis and coronary heart disease through its pro-oxidative properties. In the present study, the associations between serum ferritin and plasma oxysterol concentrations were examined in 669 eastern Finnish men. Serum ferritin concentration had statistically significant (p <.05) direct correlations with most of the measured oxysterols. In multivariate adjusted regression models, serum ferritin concentration predicted significantly the levels of 27-hydroxycholesterol (beta = 0.13, p <.001), 7alpha hydroxycholesterol (beta = 0.11, p =.005), 25-hydroxycholesterol (beta = 0.10, p =.007), 7-ketocholesterol (beta = 0.10, p =.009), and 7beta-hydroxycholesterol (beta = 0.10, p =.02). In conclusion, excess body iron, as assessed by serum ferritin, is associated with increased levels of circulating oxysterols, both of enzymatic and nonenzymatic origin, in man. PMID- 14556857 TI - Ascorbate enhances iNOS activity by increasing tetrahydrobiopterin in RAW 264.7 cells. AB - Studies on the effect of ascorbic acid on inducible nitric oxide synthase (iNOS) activity are few and diverse, likely to be dependent on the species of cells. We investigated a role of ascorbic acid in iNOS induction and nitric oxide (NO) generation in mouse macrophage cell line RAW 264.7. Although interferon- (IFN-) gamma alone produced NO end products, ascorbic acid enhanced NO production only when cells were synergistically stimulated with IFN-gamma plus Escherichia coli lipopolysaccharide (LPS). Ascorbate neither enhanced nor decreased the expression of iNOS protein in RAW 264.7 cells, in contrast to the reports that ascorbic acid augments iNOS induction in a mouse macrophage-like cell line J774.1 and that ascorbate suppresses iNOS induction in rat skeletal muscle endothelial cells. Intracellular levels of tetrahydrobiopterin (BH4), a cofactor for iNOS, were increased by ascorbate in RAW 264.7 cells. However, ascorbate did not increase GTP cyclohydrolase I mRNA, the main enzyme at the critical steps in the BH4 synthetic pathway, expression levels and activity. Sepiapterin, which supplies BH4 via salvage pathway, more efficiently enhanced NO production if ascorbate was added. These data suggest that enhanced activation of iNOS by ascorbic acid is mediated by increasing the stability of BH4 in RAW 264.7 cells. PMID- 14556859 TI - Protection conferred by Bcl-2 expression involves reduced oxidative stress and increased glutathione production during hypothermia-induced apoptosis in AK-5 tumor cells. AB - Hypothermia is known to retard mammalian cell growth, however, BC-8 cells, which have originated from AK-5 tumor after single cell cloning, were triggered into apoptotic pathway when grown at 30 degrees C. Cell death process showed typical apoptotic features like DNA fragmentation, cytochrome c release, etc. Introduction of Bcl-2 gene in BC-8 cells inhibited hypothermia-induced apoptotic process, which is ascribed to reduced ROS generation and higher glutathione production. Thus, Bcl-2 seems to control the apoptotic induction process at the level of redox regulation, in addition to its known effects at the mitochondrial dysregulation. These observations suggest that tumors, which are low in Bcl-2 expression, are sensitive to hypothermic shock and make hypothermia an interesting inducer of apoptosis in tumor cells. PMID- 14556858 TI - Superoxide and hydrogen peroxide production by Drosophila mitochondria. AB - Drosophila melanogaster is a key model organism for genetic investigation of the role of free radicals in aging, but biochemical understanding is lacking. Superoxide production by Drosophila mitochondria was measured fluorometrically as hydrogen peroxide, using its dependence on substrates, inhibitors, and added superoxide dismutase to determine sites of production and their topology. Glycerol 3-phosphate dehydrogenase and center o of complex III in the presence of antimycin had the greatest maximum capacities to generate superoxide on the cytosolic side of the inner membrane. Complex I had significant capacity on the matrix side. Center i of complex III, cytochrome c, and complex IV produced no superoxide. Native superoxide generation by isolated mitochondria was also measured without added inhibitors. There was a high rate of superoxide production with sn-glycerol 3-phosphate as substrate; two-thirds mostly from glycerol 3 phosphate dehydrogenase on the cytosolic side and one-third on the matrix side from complex I following reverse electron transport. There was little superoxide production from any site with NADH-linked substrate. Superoxide production by complex I following reverse electron flow from glycerol 3-phosphate was particularly sensitive to membrane potential, decreasing 70% when potential decreased 10 mV, showing that mild uncoupling lowers superoxide production in the matrix very effectively. PMID- 14556860 TI - Radical scavenging properties of genistein. AB - The reactivity of genistein toward reactive radical species has been investigated by means of pulse radiolysis. The values of rate constants, respectively 2.3 x 10(10) M(-1)s(-1) and 1.3 x 10(10) M(-1)s(-1) for the reaction with hydroxyl radical at pH 8.3 and 3.0, are close to diffusion limit indicating that genistein is a potent hydroxyl radical scavenger. The reactivity of genistein towards one electron oxidants has also been investigated. The rate constants k = 4.6 x 10(9) M(-1)s(-1) (pH 8.3) and 6.7 x 10(8) M(-1)s(-1) (pH 7.6) have been determined for the reaction of genistein with *N3 and Br2*- radicals, respectively. For both oxidants the rate constants at pH 3 does not exceed 10(8) M(-1)s(-1). The differences in reactivity of genistein towards the oxidants at different acidity of the solution have been assumed to arise from the acid-base equilibria of genistein. The dissociation constants for genistein (pKa: 7.2, 10.0, and 13.1) have been evaluated spectroscopically. The influence of acid-base equilibria on bond dissociation energy and ionization potential for genistein has also been investigated by means of DFT calculations. It has been concluded on the basis of these calculations that monoanionic form of genistein existing at physiological pH is more powerful radical scavenger than the neutral molecule. PMID- 14556861 TI - Role of poly(ADP-ribose)polymerase in cisplatin-induced injury in LLC-PK1 cells. AB - Acute renal failure is a dose-limiting factor during cisplatin chemotherapy. We have previously shown in rats that the hydroxyl radical scavenger edaravone reverses cisplatin-induced in vivo renal damage. In the present study, the role of poly(ADP-ribose) polymerase (PARP) in cisplatin nephrotoxicity was investigated in porcine tubular cells LLC-PK1. Cell injury was elicited by transient exposure to 500 microM cisplatin for 1 h or continuous exposure to 30 microM cisplatin for 24 h. Various hydroxyl radical scavengers reversed cell damage in a transient but not permanent model. The cell injury seemed to be necrosis and apoptosis in transient and permanent models, respectively, as assessed by TUNEL method and Annexin V stain. PARP inhibitors such as 3 aminobenzamide and benzamide inhibited cell damage in transient but not permanent model. PARP-dependent cell injury was also observed after transient exposure to hydroxyl radical-generating solution. We demonstrated for the first time the activation of PARP in renal tubular cells by transient cisplatin exposure, as determined by immunofluorescent stain with anti-poly(ADP-ribose) antibody. Moreover, ATP was depleted by transient exposure to cisplatin or hydroxyl radical, both of which were reversed by PARP inhibitors. These findings suggest that hydroxyl radical generation followed by PARP activation contributes to the necrotic cell injury caused by a transient exposure to cisplatin. PMID- 14556862 TI - MAPK pathways mediate hyperoxia-induced oncotic cell death in lung epithelial cells. AB - Cell injury and cell death of pulmonary epithelium plays an important role in the pathogenesis of acute lung injury in animals exposed to prolonged hyperoxia. The aim of this study was to decipher the molecular mechanisms modulating cell death induced by hyperoxia in lung epithelium. Cell death is thought to be either apoptotic, with shrinking phenotypes and activated caspases, or oncotic, with swelling organelles. Exposure to 95% O2 (hyperoxia) induced cell death of MLE-12 cells with cellular as well as nuclear swelling, cytosolic vacuolation, and loss of mitochondrial structure and enzyme function. Neither elevated caspase-3 activity nor phosphatidylserine translocation were detected, suggesting that in hyperoxia, MLE-12 cells die via oncosis rather than apoptosis. In addition, hyperoxia triggered a sustained activation of the transcription factor AP-1, as well as mitogen-activated protein kinase (MAPK) family members p38 and JNK. Importantly, survival of MLE-12 cells in hyperoxia was significantly enhanced when either AP-1, p38, or JNK activation was inhibited by either specific inhibitors or dominant negative DNA constructs, indicating that in lung epithelial cells hyperoxia induces a program-driven oncosis, involving AP-1, JNK, and p38 MAPK. Interestingly, hydrogen peroxide-induced oxidative apoptosis of MLE 12 cells, with a shrinking nuclear morphology and activated caspase-3 activity, is also mediated by AP-1, JNK, and p38. Therefore, our data indicate that although they have divergent downstream events, oxidative oncosis and apoptosis share upstream JNK/p38 and AP-1 pathways, which could be used as potential targets for reducing hyperoxic inflammatory lung injury. PMID- 14556863 TI - Effect of negative emotions on frequency of coronary heart disease (The Normative Aging Study). AB - Negative emotions, such as depression and anxiety, have been associated with the development of coronary heart disease (CHD). In multivariate models, negative emotions have predicted CHD outcomes, such as nonfatal myocardial infarction and CHD mortality. Few studies, however, have investigated this relation while controlling for variables associated with the metabolic syndrome or those indicative of sympathetic nervous system activity. We prospectively examined the relation between negative emotions and incident CHD in older men (mean 60.3 +/- 7.9 years) participating in the Normative Aging Study (NAS). Four hundred ninety eight men who completed the Minnesota Multiphasic Personality Inventory (MMPI) and who participated in a subsequent laboratory assessment were included in the study. All men were not on medication and free of diagnosed CHD and diabetes. Negative emotions were measured by the MMPI Welsh A scale, which is comprised of 39 items measuring symptoms of depression and anxiety. Negative emotion score, sociodemographic characteristics, health behaviors, components of the metabolic syndrome, and stress hormones were used to predict incident CHD over a 3-year follow-up period. During follow-up, 45 CHD events were observed. In unadjusted logistic regression analyses, negative emotions significantly predicted the incidence of CHD (odds ratio [OR] 1.06, 95% confidence interval [CI] 1.01 to 1.10, p = 0.02). After adjusting for potential covariates, negative emotions continued to predict the incidence of CHD (OR 1.06, 95% CI 1.01 to 1.12, p = 0.02) A linear, dose-response relation was observed (chi-square 10.8, degree of freedom 2, p = 0.005): participants who had the highest level of negative emotions experienced the greatest incidence of CHD. PMID- 14556864 TI - Temporal changes and prognostic significance of measures of heart rate dynamics after acute myocardial infarction in the beta-blocking era. AB - Previous studies have shown that indexes describing heart rate (HR) dynamics may predict subsequent deaths of patients after an acute myocardial infarction (AMI). Because beta-blocking (BB) drugs affect both mortality and HR dynamics, the prognostic power of measurements of HR dynamics may have changed in the current era of BB therapy. This study assessed the temporal changes and prognostic significance of time-domain, spectral, and fractal indexes of HR variability along with HR turbulence after an AMI among patients with optimized BB medication. SD of NN intervals, spectral indexes, the short-term fractal scaling exponent (alpha(1)), power-law slope (beta), and turbulence onset and slope were measured in 600 patients at 5 to 7 days after AMI and in 416 patients at 12 months after AMI. In the multivariate analysis, after adjusting for clinical variables, only reduced fractal HR indexes, alpha1 and beta (p <0.01 for both), turbulence onset, and slope (p <0.05 for both), measured at the convalescent phase after AMI, predicted subsequent cardiac death. All time-domain and spectral HR variability indexes and turbulence onset increased significantly during the 12 month period after AMI (p <0.001 for all), whereas the fractal indexes and turbulence slope remained unchanged. Late after AMI, reduced beta (p <0.05) and turbulence slope (p <0.01) were the only independent predictors of cardiac mortality. Traditional time-domain and spectral measurements of HR variability and turbulence onset improved significantly after AMI, whereas the fractal HR dynamics and turbulence slope remained stable. Fractal HR variability and HR turbulence retain their prognostic power in the BB era, when measured either at the convalescent or late phase after AMI. PMID- 14556865 TI - Evaluation of the association of proximal coronary culprit artery lesion location with clinical outcomes in acute myocardial infarction. AB - Impaired coronary artery blood flow and left anterior descending (LAD) artery culprit location are angiographic variables that have been associated with poorer outcomes after fibrinolytic administration in patients with acute myocardial infarction (AMI). We hypothesized that culprit lesion location in the proximal portion of the culprit artery would also be associated with poorer clinical outcomes compared with a mid or distal location. Lesion location and clinical outcomes were evaluated in 2,488 patients from the Thrombolysis In Myocardial Infarction (TIMI) 4, 10A, 10B, and 14 trials. Proximal lesions were located before or at the first major branch of the parent artery, mid lesions were between the first and the second major branches, and all other lesions were classified as distal. Proximal lesions were associated with a higher incidence of in-hospital death or recurrent AMI compared with mid or distal lesions (10.5% [n = 478] vs 6.1% [n = 1,498] vs 3.7% [n = 511], p <0.001), and they were associated with a higher rate of in-hospital death (6.7% [n = 478] vs 3.2% [n = 1,498] vs 2.5% [n = 511], p = 0.001). In a multiple logistic regression model adjusting for TIMI flow grade, age, gender, and pulse, the planimetered distance from the ostium to the LAD culprit lesion was associated with 30-day death or recurrent AMI (odds ratio 0.79 per centimeter increase in distance down the artery, p = 0.01). Proximal culprit lesion location is associated with an increased risk of adverse outcomes after fibrinolytic administration, which is likely due to a larger area of subtended myocardium. In patients with a LAD culprit lesion, proximal lesion location is a multivariate correlate of adverse outcomes even after adjustment for coronary blood flow and other covariates. PMID- 14556866 TI - Bivalirudin provides increasing benefit with decreasing renal function: a meta analysis of randomized trials. AB - Chronic kidney disease is associated with an increased risk of ischemic and bleeding events after percutaneous coronary intervention (PCI). The direct thrombin inhibitor bivalirudin reduces these combined events. We sought to assess whether this benefit was influenced by renal function. A meta-analysis of 3 randomized trials (n = 5,035) comparing bivalirudin with heparin during PCI, stratified by estimated creatinine clearance using the Cockcroft-Gault equation (>90 [n = 1,578], 90 to 60 [n = 2,163], 59 to 30 [n = 1,255], and <30 ml/min [n = 39]), was conducted. The composite end points of death, myocardial infarction or revascularization, hemorrhage, and combined ischemic or bleeding events were assessed. A common odds ratio for each creatinine clearance strata was estimated with a random-effects model. The interaction between renal impairment and benefit from bivalirudin was assessed. Adverse ischemic and bleeding events increased with decreasing renal function. The relative benefit of bivalirudin with respect to ischemic and bleeding events was maintained within each stratum. The absolute benefit in terms of ischemic and bleeding complications increased with decreasing creatinine clearance (normal 2.2%, mild 5.8%, moderate 7.7%, severe 14.4%; p trend <0.001, interaction p = 0.044). Renal dysfunction remains a prevalent risk factor for ischemic and bleeding events in patients who undergo PCI. Bivalirudin provides greater absolute benefit in patients with impaired renal function. PMID- 14556867 TI - Comparison of inflammatory markers in patients with diabetes mellitus versus those without before and after coronary arterial stenting. AB - Patients with diabetes are at increased risk for adverse events after coronary stenting, perhaps reflecting a pro-inflammatory state. To characterize the inflammatory response to coronary stenting in patients with and without diabetes, blood samples were obtained from 75 patients before stenting and 10 minutes, 1 hour, and 24 hours later. C-reactive protein (CRP, microg/ml), interleukin (IL)-6 (pg/ml), IL-1 receptor antagonist (pg/ml), and soluble CD40 ligand (ng/ml) were assayed in each sample by enzyme-linked immunosorbent assay. Concentration changes after stenting were identified by repeated-measures analysis of variance. Multivariate analysis was performed to delineate independent predictors of increased concentrations of inflammation markers. Overall, 88% of patients had acute coronary syndromes; 36% had elevated markers of cardiac injury. The preprocedural concentrations of CRP in those with diabetes were more than twice as high as those in patients without diabetes. Two independent predictors of elevated preprocedural CRP concentrations were diabetes (odds ratio 3.95, 95% confidence interval 1.17 to 13.4) and a cardiac marker-positive acute coronary syndrome (odds ratio 3.70, 95% confidence interval 1.22 to 11.2). Preprocedural concentrations of IL-6, IL-1 receptor antagonist, and soluble CD40 ligand tended to be greater in patients with diabetes. The increase in CRP after stenting was much greater for patients without diabetes compared with that in patients with diabetes such that the apparent intensity of inflammation after 24 hours was similar in those with and without diabetes. Thus, patients with and without diabetes exhibit different inflammatory responses to stenting, reflecting the lower preprocedural inflammation in those without diabetes versus those with diabetes. PMID- 14556868 TI - Incidence, predictors, and prognostic implications of bleeding and blood transfusion following percutaneous coronary interventions. AB - Bleeding related to percutaneous coronary intervention (PCI) occurs relatively frequently. We retrospectively investigated the incidence, predictors, and prognostic impact of periprocedural bleeding and transfusion in 10,974 patients who underwent PCI. Bleeding definitions were based on Thrombolysis In Myocardial Infarction (TIMI) criteria: (1) major bleeding (n = 588; 5.4%): if patients had a hemorrhagic stroke or if hematocrit decreased >15 points or by 10 to 15 points with clinical bleeding; (2) minor bleeding (n = 1,394; 12.7%): if hematocrit decreased <10 points with clinical bleeding or by 10 to 15 points without clinical bleeding; and (3) no bleeding (n = 8,992; 81.9%): if hematocrit decreased <10 points without clinical bleeding. Patients with major bleeding were older than patients with minor or no bleeding (67.8 +/- 11 vs 65.9 +/- 11 vs 63.6 +/- 11 years, respectively; p <0.001) and more often experienced intraprocedural complications, such as emergency use of an intra-aortic balloon pump (13.6% vs 6.5% vs 2.3%, respectively; p <0.001). Multivariate logistic regression analysis identified the use of an intra-aortic balloon pump (odds ratio [OR] 3.0, p <0.0001), procedural hypotension (OR 2.9, p <0.001), and age >80 years (OR 1.9 compared with age <50 years, p = 0.001) as the strongest predictors for major bleeding. Patients who had major bleeding had higher in-hospital and 1-year mortality compared with patients with minor or no bleeding. Bleeding was an independent predictor of in-hospital death. Thus, periprocedural major bleeding occurs relatively frequently and is associated with adverse outcomes. Patients >80 years of age who experience intraprocedural complications are at particularly high risk. PMID- 14556869 TI - Intravascular ultrasound-guided directional coronary atherectomy for unprotected left main coronary stenoses with distal bifurcation involvement. AB - Stent implantation in unprotected left main coronary artery (LMCA) bifurcation lesions may improve procedural and late clinical outcomes. However, concerns regarding stent-related complications, such as stent jail, subacute thrombosis, and in-stent restenosis remain. Optimal debulking by directional coronary atherectomy (DCA) with intravascular ultrasound (IVUS) guidance may be effective in this complex lesion subset, but this strategy has not yet been established. Our objective was to evaluate the safety and efficacy of IVUS-guided DCA for unprotected LMCA stenoses with distal bifurcation involvement. A total of 67 consecutive patients were included in this study and procedural success was achieved in all cases. Two cardiac deaths (2.9%) were noted and 3 patients (4.5%) underwent repeat angioplasty during hospitalization. There was no Q-wave myocardial infarction or emergency bypass surgery. Non-Q-wave myocardial infarction (creatine kinase-MB >3 times normal) occurred in 13.4% of patients. Stent implantation was necessary in 17 cases (25.4%) to achieve an optimal result. IVUS showed an improved lumen cross-sectional area and a low plaque burden in the LMCA after intervention. All-cause mortality, angiographic restenosis, and the target lesion revascularization rates at 6 months were 7.4%, 23.8%, and 20.0%, respectively. With IVUS guidance, aggressive DCA can be performed safely in unprotected LMCA bifurcation lesions, and optimal angiographic and IVUS results can be achieved with low residual plaque burden, which leads to a low restenosis rate. Optimal lesion debulking by DCA does not necessarily need adjunctive stenting in this specific anatomic subset. PMID- 14556870 TI - Efficacy and safety of sustained-release propafenone (propafenone SR) for patients with atrial fibrillation. AB - The objective of this randomized, double-blind, placebo-controlled clinical trial was to test the efficacy and safety of a new sustained-release preparation of the antiarrhythmic drug propafenone (propafenone SR) in reducing the frequency of symptomatic arrhythmia recurrences in patients with atrial fibrillation (AF). Patients with a history of symptomatic AF who were in sinus rhythm were randomly assigned to receive placebo or propafenone SR 425, 325, or 225 mg, all twice daily. Recurrent symptomatic arrhythmias were documented using transtelephone electrocardiographic monitoring. Electrocardiograms were reviewed by an event committee that was blinded to treatment assignment. In the primary efficacy analysis, propafenone SR significantly lengthened the time to first symptomatic atrial arrhythmia recurrence at all 3 doses compared with placebo as assessed by log-rank test: propafenone SR 425 mg twice daily versus placebo twice daily, p <0.001; 325 mg twice daily versus placebo twice daily, p <0.001; and 225 mg twice daily versus placebo twice daily, p = 0.014. The median time to recurrence was 41 days in the placebo twice daily group, >300 days in the propafenone SR 425-mg group, 291 days in the 325-mg group, and 112 days in the 225-mg group. Adverse effects leading to withdrawal were higher in the propafenone SR 425-mg twice daily group than in any other group. Thus, propafenone SR has important and statistically significant antiarrhythmic effects in patients with AF. PMID- 14556871 TI - Radiofrequency catheter ablation of septal accessory pathways in the pediatric age group. AB - Radiofrequency catheter ablation (RCA) of septal accessory pathways may be technically challenging in children due to the risk of inadvertent atrioventricular (AV) block in the setting of small cardiac dimensions. Outcomes were reviewed for all patients aged < or =19 years with manifest and concealed septal accessory pathways undergoing RCA since 1990 at a single institution. One hundred forty-five procedures were performed in 127 patients (mean age 11.6 years). The number of studies according to accessory pathway location were: anteroseptal (n = 36), midseptal (n = 20), mouth of coronary sinus (n = 40), middle cardiac vein (n = 6), right posteroseptal (n = 21), and left posteroseptal (n = 22). Ablation was deferred for 9 patients (6 anteroseptal and 3 midseptal) in favor of additional pharmacologic trials. Acute success rates for targeted accessory pathways were: anteroseptal (96%), midseptal (94%), mouth of coronary sinus (88%), middle cardiac vein (100%), right posteroseptal (100%), and left posteroseptal (96%). Recurrence rates during follow-up were: anteroseptal (14%), midseptal (12%), mouth of coronary sinus (3%), right posteroseptal (4%), and left posteroseptal (4%). Permanent second or third degree AV block occurred in 4 of 136 RCA attempts (3%), involving 2 anteroseptal and 2 midseptal pathways. In 3 of these 4 cases, a high probability of block was anticipated from prior ablation efforts, prompting pacemaker insertion before or in conjunction with RCA. Thus, in the pediatric age group, acute RCA success rates for septal accessory pathways can exceed 90%. The risks of AV block and accessory pathway recurrence are most relevant to anteroseptal and midseptal pathways. These data may be factored into patient selection and the decision whether to ablate. PMID- 14556873 TI - Long-term prognostic implication of extracardiac vascular disease in patients undergoing percutaneous coronary intervention. AB - Patients with extracardiac vascular disease were identified from 2,372 consecutive percutaneous coronary intervention (PCI) cases performed between 1997 and 2001. After multivariate adjustment, we found the presence of extracardiac vascular disease to be associated with a significantly higher risk for late mortality (hazard ratio [HR] 1.4, 95% confidence interval [CI] 1.0 to 2.0, p = 0.029). When extracardiac vascular disease was separated into cerebrovascular disease and peripheral vascular disease, cerebrovascular disease was less common but was associated with a trend towards worse survival. PMID- 14556872 TI - Relation of thoracic aortic and aortic valve calcium to coronary artery calcium and risk assessment. AB - Aortic calcium, aortic valve calcium (AVC), and coronary artery calcium (CAC) have been associated with cardiovascular event risk. We examined the prevalence of thoracic aortic calcium (TAC) and AVC in relation to the presence and extent of CAC, cardiovascular risk factors, and estimated risk of coronary heart disease (CHD). In 2,740 persons without known CHD aged 20 to 79 years, CAC was assessed by electron beam- or multidetector-computed tomography. We determined the prevalence of TAC and AVC in relation to CAC, CHD risk factors, and predicted 10 year risk of CHD. A close correspondence of TAC and AVC was observed with CAC. TAC and AVC increased with age; by the eighth decade of life, the prevalence of TAC was similar to that of CAC (>80%), and 36% of men and 24% of women had AVC. Age, male gender, and low-density lipoprotein cholesterol were directly related to the likelihood of CAC, TAC, and AVC; higher diastolic blood pressure and cigarette smoking additionally predicted CAC. Body mass index and higher systolic and lower diastolic blood pressures were also related to TAC, and higher body mass index and lower diastolic blood pressure were related to AVC. Calculated risk of CHD increased with the presence of AVC and TAC across levels of CAC. TAC and AVC provided incremental value over CAC in association with the 10-year calculated risk of CHD. If longitudinal studies show an incremental value of aortic and aortic valve calcium over that of CAC for prediction of cardiovascular events, future guidelines for risk assessment incorporating CAC assessment may additionally incorporate the measurement of aortic and/or aortic valve calcium. PMID- 14556874 TI - Frequency and prognosis of emergency coronary artery bypass grafting after percutaneous coronary intervention for acute myocardial infarction. AB - We evaluated the frequency and prognosis of emergency coronary artery bypass grafting (CABG) after percutaneous coronary intervention (PCI) for acute myocardial infarction in a large, multicenter registry of contemporary PCI. In this study, emergency CABG occurred in 2% of cases, and was associated with high in-hospital mortality (20%) and with a high incidence of stroke (8%), renal failure requiring dialysis (8.3%), and bleeding (63.3%). PMID- 14556875 TI - Incidence and mechanism of late stent malapposition after phosphorus-32 radioactive stent implantation. AB - Late stent malapposition is a potential complication of intracoronary brachytherapy. The aim of our study was to determine the incidence and mechanism of late stent malapposition after implantation of phosphorus-32 radioactive stents. We analyzed 159 patients for de novo lesions after the implantation of phosphorus-32 radioactive stents. There were 15 late stent malappositions. The incidence of malapposition was higher in patients who received Hot-Ends Isostents. External elastic membrane expansion greater than plaque increase in malapposed segments was observed. Late stent malapposition is caused by a localized increase in external elastic membrane that is greater than the increase in plaque area; this is believed to be a dose-dependent phenomenon because it was more common with Hot-Ends Isostents. PMID- 14556876 TI - Impact of extracardiac vascular disease on acute prognosis in patients who undergo percutaneous coronary interventions (data from the Blue Cross & Blue Shield of Michigan Cardiovascular Consortium [BMC2]). AB - Extracardiac vascular disease is associated with an increased risk of in-hospital mortality and other complications after coronary interventions, independent from other co-morbidities and baseline characteristics. The underlying cause of this significant association is unclear, but it warrants further investigation in an attempt to improve outcome in this high-risk cohort. PMID- 14556877 TI - Effect of pravastatin on coronary plaque volume. AB - A volumetric analysis by 3-dimensional intravascular ultrasound revealed that lipid-lowering therapy with pravastatin significantly reduced coronary plaque volume. The changes in plaque volume were inversely correlated with the changes in plasma levels of high-density lipoprotein cholesterol but not with changes in levels of total cholesterol or low-density lipoprotein cholesterol. PMID- 14556878 TI - Effect of tirofiban before primary angioplasty on initial coronary flow and early ST-segment resolution in patients with acute myocardial infarction. AB - Sixty-one patients with ST elevation acute myocardial infarction were randomized to receive open-label tirofiban in the emergency room before primary angioplasty versus glycoprotein IIb/IIIa inhibitors administered after initial coronary angiography. Early administration of tirofiban before primary angioplasty resulted in nonsignificant improvement in initial coronary flow (Thrombolysis In Myocardial Infarction trial grade 2 or 3 flow, 39% vs 27%, p >0.20). Patients receiving early tirofiban treatment were more likely to achieve complete (>70%) ST-segment resolution at 90 minutes (69% vs 44%, p = 0.07). PMID- 14556879 TI - Comparison of creatine kinase elevation and outcome of comparison of percutaneous coronary intervention for saphenous vein graft in-stent restenosis versus de novo stenosis. AB - Percutaneous coronary intervention of saphenous vein grafts is associated with distal embolization. We aimed to compare potential differences in patients undergoing percutaneous coronary intervention for in-stent restenosis with de novo saphenous vein graft lesions. Myocardial necrosis was associated with higher mortality regardless of lesion type. PMID- 14556880 TI - Social and psychosocial influences on inflammatory markers and vascular function in civil servants (the Whitehall II study). AB - Social position and psychosocial factors are associated with coronary disease, but the underlying pathophysiologic mechanisms remain unclear. In a sample of 283 nonsmokers, we found that social position was inversely associated with interleukin-6 and C-reactive protein and that participants with mild depression had impaired endothelial function. PMID- 14556881 TI - Efficacy and impact of monophasic versus biphasic countershocks for transthoracic cardioversion of persistent atrial fibrillation. AB - This report compares the cumulative efficacy of cardioversion and skeletal muscle injury after either damped sine wave monophasic or truncated exponential biphasic transthoracic cardioversion of persistent atrial fibrillation. The trial sought to refute the null hypothesis of therapeutic equivalence between monophasic and biphasic waveforms. Results of the study reveal similar cumulative efficacy of waveforms and greater levels of skeletal muscle damage when patients are younger and male, and when monophasic waveforms are used. PMID- 14556882 TI - Bidirectional ventricular tachycardia and channelopathy. AB - Based on similarity of electrocardiographic features, bidirectional ventricular tachycardia has been considered a variant of long QT syndrome. Genes causing long QT syndrome were used as candidate genes in 4 patients with bidirectional ventricular tachycardia. In 2 patients, we identified a common low penetrance HERG allele (R1047L) with an intermediate biophysical phenotype. PMID- 14556883 TI - QT prolongation and Torsades de Pointes in patients infected with human immunodeficiency virus and treated with methadone. AB - Four patients infected with human immunodeficiency virus receiving antiretroviral treatment and high doses of methadone (>200 mg/day) presented with several syncopal episodes. A significant prolongation of the QTc interval was detected in all of them, and in 3 patients, > or =1 episode of Torsades de Pointes was recorded. The sequence of events in these cases suggests that high doses of methadone caused QT prolongation and provided the substrate for syncope and Torsades de Pointes. PMID- 14556884 TI - Relation between microvolt-level T-wave alternans and cardiac sympathetic nervous system abnormality using iodine-123 metaiodobenzylguanidine imaging in patients with idiopathic dilated cardiomyopathy. AB - We examined the relation between microvolt-level T-wave alternans and cardiac sympathetic nervous system abnormality using iodine-123 metaiodobenzylguanidine imaging in patients with idiopathic dilated cardiomyopathy. Our results strongly indicate that cardiac sympathetic denervation and accelerated sympathetic nervous activity play important roles in the presence of microvolt-level T-wave alternans in patients with idiopathic-dilated cardiomyopathy. PMID- 14556885 TI - Usefulness of chronotropic incompetence to dipyridamole in predicting myocardial perfusion defects in heart transplant recipients. AB - The aim of this report was to assess the relation between heart rate response to dipyridamole infusion and perfusion defects at quantitative sestamibi single photon emission computed tomographic imaging. We demonstrated in 166 heart transplant recipients that chronotropic incompetence to dipyridamole is the only significant and independent predictor of perfusion defects. PMID- 14556886 TI - Prospective study of bacteremia after cardiac catheterization. AB - Nine hundred sixty consecutive cardiac catheterization procedures were studied prospectively for the presence of periprocedural bacteremia. Overall, among 960 procedures, only 4 were associated with clinically significant bacteremia. All 4 were related to the intravenous line and none to the cardiac procedure itself. Clinically nonsignificant bacteremias were correlated with procedural duration, multiple skin punctures, use of multiple balloons, and obesity. PMID- 14556887 TI - Comparison of acute hemodynamic effects of aerosolized and intravenous iloprost in secondary pulmonary hypertension in children with congenital heart disease. AB - Both aerosolized and intravenous infusion of iloprost caused a significant decrease in mean pulmonary artery pressure and pulmonary vascular resistance. Although intravenous infusion caused a large decrease in mean systemic arterial pressure, this was only slightly affected by aerosolized iloprost. Aerosolized iloprost caused a significant decrease in the pulmonary-to-systemic vascular resistance ratio; however, intravenous infusion did not cause a prominent decrease in this ratio. PMID- 14556889 TI - Influence of residual stenosis after coronary stent implantation on development of restenosis and stent thrombosis. PMID- 14556891 TI - A theoretical study of effects of cytosolic Ca2+ oscillations on activation of glycogen phosphorylase. AB - Taking into account the Ca(2+)-stimulated degradation of inositol 1,4,5 trisphosphate (IP(3)) by a 3-kinase, we have theoretically explored the effects of both simple and complex Ca(2+) oscillations on the regulation of a phosphorylation-dephosphorylation cycle process involved in glycogen degradation by glycogen phosphorylase a-form, respectively. For the case of simple Ca(2+) oscillations, the roles of cytosolic Ca(2+) oscillations in the regulation of active phosphorylase depend upon the maximum rate of IP(3) degradation by the 3 kinase, V(M5). In particular, the smaller the values of V(M5) are, the lower the effective Ca(2+) threshold for the activation of glycogen phosphorylase will be. For the case of complex Ca(2+) oscillations, the average level of fraction of active phosphorylase is nearly independent from the level of stimulation increasing in the bursting oscillatory domain. Both simple and complex Ca(2+) oscillations can contribute to increase the efficiency and specificity of cellular signalling, and some theoretical results of activation of glycogen phosphorylase regulated by Ca(2+) oscillations are close to the experimental results for gene expression in lymphocytes. PMID- 14556892 TI - Gating kinetics of potassium channel in rat dorsal root ganglion neurons analyzed with fractal model. AB - The kinetics of ion channels have been widely modeled as a Markov process. In these models it is assumed that the channel protein has a small number of discrete conformational states and kinetic rate constants connecting these states are constant. To study the gating kinetics of voltage-dependent K(+) channel in rat dorsal root ganglion neurons, K(+) channel current were recorded using cell attached patch-clamp technique. The K(+) channel characteristic of kinetics were found to be statistically self-similar at different time scales as predicted by the fractal model. The fractal dimension D for the closed times and for the open times depend on the pipette potential. For the open and closed times of kinetic setpoint, it was found dependent on the applied pipette potential, which indicated that the ion channel gating kinetics had nonlinear kinetic properties. Thus, the open and closed durations, which had the voltage dependence of the gating of this ion channel, were well described by the fractal model. PMID- 14556893 TI - Signal processing times in neutrophil activation: dependence on ligand concentration and the relative phase of metabolic oscillations. AB - Intracellular NAD(P)H oscillations exhibited by polarized neutrophils display congruent with 20 s periods, which are halved to congruent with 10 s upon stimulation with chemotactic peptides such as FNLPNTL (N-formyl-nle-leu-phe-nle tyr-lys). By monitoring this frequency change, we have measured accurately the time interval between stimulus and metabolic frequency changes. A microscope flow chamber was designed to allow rapid delivery of FNLPNTL to adherent cells. Using fluorescein as a marker, we found delivery to be complete and stable throughout the chamber within approximately 400 ms. Peptides were injected into the chamber at concentrations ranging from 10(-6) to 10(-9) M. Injections also varied with respect to the relative phase of a cell's NAD(P)H oscillations. The time interval between injection of 10(-6) M FNLPNTL and the acquisition of congruent with 10 s period metabolic oscillations was found to be 12.2+/-3.3 s when injections occurred at the NAD(P)H oscillation peak whereas the lag time was 22.5+/-4.8 s when coinciding with a trough. At 10(-8) M FNLPNTL, lag times were found to be 26.1+/-5.2 and 30.5+/-7.3 s for injections at NAD(P)H peaks and troughs, respectively. FNLPNTL at 10(-9) M had no effect on metabolic oscillations, consistent with previous studies. Our experiments show that the kinetics of transmembrane signal processing, in contrast to a simple transmembrane chemical reaction, can depend upon both ligand dose and its temporal relationship with intracellular metabolic oscillations. PMID- 14556894 TI - Excited state charge-transfer dynamics study of poplar plastocyanin by ultrafast pump-probe spectroscopy and molecular dynamics simulation. AB - We have applied ultrafast pump-probe spectroscopy to investigate the excited state dynamics of the blue copper protein poplar plastocyanin, by exciting in the blue side of its 600-nm absorption band. The decay of the charge-transfer excited state occurs exponentially with a time constant of approximately 280 fs and is modulated by well visible oscillations. The Fourier transform of the oscillatory component, besides providing most of the vibrational modes found by conventional resonance Raman, presents additional bands in the low frequency region modes, which are reminiscent of collective motions of biological relevance. Notably, a high frequency mode at approximately 508 cm(-1), whose dynamics are consistent with that of the excited state and already observed for other blue copper proteins, is shown to be present also in poplar plastocyanin. This vibrational mode is reproduced by a molecular dynamics simulation involving the excited state of the copper site. PMID- 14556895 TI - An NMR and circular dichroism study of the interaction of thiocyanate with human and cross-linked hemoglobin: identification of Lys-alpha-99 as a possible dissociation linked binding site. AB - The interaction of thiocyanate with human native and cross-linked oxyhemoglobin (oxyHb), and methemoglobin (metHb) has been investigated by optical spectroscopy, circular dichroism (CD) and nuclear spin lattice relaxation rate measurements. The interaction of thiocyanate anion with human hemoglobin has been investigated by NMR measurements of the nuclear spin lattice relaxation rate of N(15) labeled thiocyanate in the presence of cyanomethemoglobin and cross-linked cyanomethemoglobin. Results show that thiocyanate is located approximately 8.9 and 6.2 A away from the heme group in cyanomethemoglobin and cross-linked cyanomethemoblobin, respectively. These results are consistent with the binding of SCN(-) at the lys-alpha-99 in the unmodified hemoglobin. Since this site is blocked in the cross-linked hemoglobin, the binding site is different. Results show that one mole of SCN(-) is binding to one mole of oxyhemoglobin suggesting that binding at the lys-alpha-99 is linked to dissociation of the hemoglobin tetramer into dimers due to its location at the alpha(1)beta(2) interface. Circular dichroism studies show that the interaction of thiocyanate with oxyHb decreases the optical rotation at 240 nm indicating a conformational change of the protein, which influences the electronic transitions of a number of peptide bonds or (and) a few aromatic side chains. PMID- 14556896 TI - Kinetic characterization of TAR RNA-Tat peptide and neomycin interactions by acoustic wave biosensor. AB - The kinetics of binding of short Tat peptides and an aminoglycoside molecule to the human immunodeficiency virus-type 1(HIV-1) TAR RNA and to a bulge mutant analogue (MTAR) is studied in a biosensor format by monitoring the time course of the response in a series resonance frequency, using an acoustic wave biosensor. Association and dissociation rate constants are evaluated by fitting the experimental data to a simple 1:1 (Langmuir) model. Kinetic rate and equilibrium dissociation constants show that MTAR-peptide complexes are subject to a higher dissociation rate and are less stable compared to the corresponding TAR-peptide complexes. In addition, longer peptides display enhanced discrimination ability than a shorter peptide according to the equilibrium dissociation constants evaluated using this technique. K(D) values for TAR-Tat vs. MTAR-Tat complexes are 2.6 vs. 3.8 microM for Tat-12, 0.87 vs. 4.3 microM for Tat-18 and 0.93 vs. 1.6 microM for Tat-20. The equilibrium dissociation constant for TAR-neomycin complex is 12.4 microM and it is comparable to the values obtained from non biosensor type assays. These findings are in parallel with those cited in the literature and the results from this study underline the potential of the acoustic wave sensor for detailed biophysical analysis of nucleic acid-ligand binding. PMID- 14556897 TI - New pinch-porphyrin complexes with quantum mixed spin ground state S=3/2,5/2 of iron (III) and their catalytic activity as peroxidase. AB - New complexes of the pinch-porphyrin family were obtained from the dimethylester of (proto-, meso-, and deutero-porphyrinato)iron(III) with the ligand [N,N'-bis pyridin-2-ylmethyl-propane-1,3-diamine] 1-3 and with the ligand [N-pyridin-2 ylmethyl-N'-[3-[(pyridin-2-ylmethyl)-amino]-propyl]-propane-1,3-diamine] 4-6. The UV/VIS studies of 1-6 indicate an increase in the distortion of the ligand field excited state. The 1H NMR spectra of 1-6 at RT and over the range 223-328 K show iron(III)-complexes with quantum mixed spin state (qms) S=5/2, S=3/2. The chemical shifts of the meso protons are consistent with qms state S=3/2, S=5/2, where the S=3/2 spin state is lowest in energy. For methyl-heme the chemical shifts are also consistent with a qms state but now the S=5/2 ground state is lowest in energy. ESR spectra of 1-6 show two different species, B and C, of iron(III) with qms, S=5/2, S=3/2 consistent with the 1H NMR results. Species B with 70% of S=5/2 and species C with 72.5% of S=3/2. The catalytic activity as peroxidase of 1-6 was quantified by guaiacol test; their theoretical maximum rate constants were k(cat) approximately 10(2)-10(3) M(-1) s(-1). A quantitative empirical correlation is found: the higher the 32 spin contribution to the qms state and the higher proportion of this species into the samples, the higher the peroxidase activity. Such a correlation was also obtained for pinch-porphyrins already reported. PMID- 14556898 TI - An electrochemical investigation of effect of ATP on hemoglobin. AB - The titratable potentiometric response of hemoglobin (Hb) induced by adenosine-5' triphosphate (ATP) is observed. The concentration-dependent effect of ATP on the anaerobic redox reaction of the protein at pH 7.0 reflects that ATP will induce stabilization of the reduced state and destabilization of the R-like (met Fe(III)) state of the metHb, when ATP concentration is lower than 3.0 mM. But when ATP concentration is between 4 and 7 mM, shift of the oxidation potential may also be observed. With reference to the study of adenosine, adenosine-5' monophosphate, adenosine-5'-diphosphate and 2,3-diphosphoglycerate, the allosteric effect of ATP on Hb is discussed extensively. This study has given an electrochemical approach to the investigation of effect of ATP, an in vivo allosteric effector, on Hb in the physiological concentration range. PMID- 14556899 TI - Long-range correlations in the helix free energy distribution in DNA. AB - In this paper we explore the free energy distribution in the helical form of DNA using the genome of the virus Rickettsia prowazekii Madrid E as an example. The genome of this organism has been determined by Andersson et al. (Nature 396 (1998) 133) and is available on the World Wide Web (www.tigr.org). Using the helix statistical weights based on nearest-neighbor base pairs of SantaLucia (Proc. Natl. Acad. Sci. USA 95 (1998) 1460), we calculate the free energy in consecutive blocks of m base pairs in the DNA sequence and then construct the free energy distribution for these values. Using the maximum-entropy method we can fit the distribution curves with a function based on the moments of the distribution. For blocks containing 10-20 base pairs the distribution is slightly skewed and we require four moments to accurately fit the function. For blocks containing 100 base pairs or more, the distribution is well approximated by a Gaussian function based on the first two moments of the distribution. We find that the free energy distribution for m=20 can be reproduced using random sequences that have the local (singlet, doublet or triplet) statistics of Rickettsia. However, for much larger blocks, for example m=500, the width of the free energy distribution based on the actual Rickettsia genome is broader by almost a factor of 3 than the distributions based on random local statistics. We find that the distribution functions for the C or G content in blocks of m base pairs have almost the same behavior as a function of block size as do the free energy distributions. In order to duplicate the width of the distribution functions based on the actual Rickettsia sequence, we need to introduce tables (matrices) that correlate the states of consecutive blocks hundreds of base pairs long. This indicates that correlations on the order of the number of base pairs contained in the average gene are required to give the actual widths for either the C or G content or the helix free energy distributions. Above a certain m value, the distributions for larger m can be accurately expressed in terms of the distribution functions for smaller m. Thus, for example, the distribution for m=5000 can be expressed in terms of the generating function for m=1000. PMID- 14556900 TI - Kinetics of heat- and acidification-induced aggregation of firefly luciferase. AB - The general approach to analysis of the kinetics of protein aggregation registered by the turbidimetric method has been elaborated. The terminal part of the kinetic curves is analyzed using a theoretical equation connecting the derivative of the apparent absorbance (A) with respect to time (dA/dt) and A (t is time). This analysis allows the limiting value of A at t--> infinity (A(lim)) and the order of aggregation with respect to protein (n) to be calculated. Approach proposed was applied to analysis of thermal and acidification-induced aggregation of firefly luciferase. In both cases the A(lim) value is a linear function of the protein concentration. The terminal part of the kinetic curves of thermal aggregation follows the first-order kinetics (n=1), whereas the kinetics of acidification-induced aggregation are characterized by the value of n higher than unity (n=1.29). The mechanism of nucleation-dependent aggregation has been discussed. PMID- 14556901 TI - MD simulation of a plastocyanin mutant adsorbed onto a gold surface. AB - MD simulation of plastocyanin, an electron transfer protein, adsorbed onto a gold surface, has been performed for 10 ns. Starting from the crystallographic structure of a poplar plastocyanin mutant engineered with the insertion of a disulfide bridge, the protein has been anchored to a gold substrate modeled by a cluster of three layers in the Au<111> configuration. A number of significant structural and dynamical properties of the protein molecule, covalently bound through either one or two sulfur atoms to the gold surface, has been extracted and compared with those of the free protein. Attention has been paid to investigate the dynamical aspects putatively related to the electron transfer process. In particular, the cross-correlation function between specific active site vibrations and all the other protein atom motions and the principal component analysis have been calculated in order to put into evidence dynamical correlation of some functional relevance. The results are discussed also in connection with related experiments. PMID- 14556902 TI - Diffusion and partition of solutes in cartilage under static load. AB - We describe experimental apparatus, methodology and mathematical algorithms to measure diffusion and partition for typical small ionic solutes and inulin (a medium size solute) in statically loaded cartilage. The partition coefficient based on tissue water (K(H(2)O)) of Na(+) increased from 1.8 to 4.5 and for SO(4)(-2) decreased from 0.5 to 0.1, when the applied pressure was raised from zero to 22 atm K(H(2)O) of inulin decreased from 0.3 to 0.05, for an increase in pressure from zero to 11 atm. Our theoretical interpretation of the results is that the partition coefficient can be expressed as a function of fixed charge density (FCD) for both loaded and unloaded cartilage. The partition coefficient shows good agreement with the ideal Gibbs-Donnan equilibrium, particularly when FCD is based on extrafibrillar water (EFW). The diffusion coefficients, D also decreased with an increase in applied pressure; raising the pressure from 0 to 22 atm resulted in the following changes in the values of D: for Na(+) from 2.86 x 10(-6) to 1.51 x 10(-6) cm(2)/s, for SO(4)(-2) from 1.58 x 10(-6) to 7.5 x 10(-7) cm(2)/s, for leucine from 1.69 x 10(-6) to 8.30 x 10(-7) cm(2)/s and for inulin from 1.80 x 10(-7) to 3.30 x 10(-8) cm(2)/s. For the three small solutes (two charged and one neutral) the diffusion coefficient D is highly correlated with the fraction of fluid volume in the tissue. These experimental results show good agreement with the simple model of Mackie and Meares: hence solute charge does not affect the diffusion of small solutes under load. For inulin D & K show some agreement with a modified Ogston model based on two major components, viz., glycosaminoglycans (GAG) and core protein. We conclude that the changes in the partition and diffusion coefficients of small and medium size solutes in statically loaded cartilage can be interpreted as being due to the reduction in hydration and increase in FCD. The change in the latter affects the partition of small ionic solutes and the partition and diffusion of larger molecules. Our results throw light on the ionic environment of chondrocytes in loaded cartilage as well as on the transport of solutes through the matrix. PMID- 14556903 TI - Protein hydration and the huge electrostriction. AB - Recent experiments indicated the existence of hydration shells about biomolecules with densities markedly higher than that of bulk water. The compression is due to the pull of the dipoles of H(2)O molecules, necessary to achieve the thermodynamic equilibrium, from bulk water into the high field (approx. 10(9) V/m) region at the surface of the protein molecule. The electric field values at the surfaces of the biomolecules are calculated on the basis of the known densities. The reverse calculation of the limiting density values on the basis of known electric field distributions is performed, too. The results compare favourably with experiment. PMID- 14556905 TI - A theory on the origin of cooperativity in DNA renaturation kinetics. AB - Theoretical proof for the existence of cooperativity (experimental evidences have already reported in our earlier work: Biochem. Biophys. Res. Commun., 293 (2002) 870) in the phenomenon of DNA renaturation has been obtained using a generalized birth-death master equation approach. Results have shown that no special searching mechanism (i.e. a facilitated mechanism) was necessary for complementary DNA strands to find their correct-contacts but searching by a 'random jump' method was enough to explain not only the observed cooperativity phenomenon but also the magnitude of correct-contact-forming rate constant. Impact of cooperativity phenomenon on length of primers and their specificity in polymerase chain reaction has also been discussed. PMID- 14556904 TI - Synthesis, solution structure analysis and antibody binding of cyclic epitope peptides from glycoprotein D of Herpes simplex virus type I. AB - Two cyclic peptides with a thioether bond have been synthesised corresponding to the 9-22 (9LKMADPNRFRGKDL(22)) sequence of glycoprotein D (gD-1) of Herpes simplex virus. The role of the secondary structure in protein-specific monoclonal antibody recognition was investigated. The sequence selected for this study comprises a strongly antigenic site adopting a beta-turn at residues 14Pro (15)Asn. Thioether bond was formed between the free thiol group of cysteine or homocysteine inserted in position 11 and the chloroacetylated side chain of lysine in position 18. We report here the preparation of cyclic peptides containing Cys or Hcy in position 11, differing only in one methylene group. The linear precursor peptides were synthesised by Boc/Bzl strategy on MBHA resin, and the cyclisation was carried out in alkaline solution. The secondary structure of the peptides was studied by CD, FT-IR and NMR spectroscopy. The CD and FT-IR data have revealed fundamental changes in the solution conformation of the two compounds. The CH(2) group difference significantly resulted in the altered turn structure at the 12Ala and 13Asp as identified by NMR spectroscopy. The antibody binding properties of the cyclopeptides studied by gD-specific monoclonal antibody (A16) in direct and competition enzyme-linked immunosorbent assay (ELISA) were also not the same. We found that peptide LK[HcyADPNRFK]GKDL exhibited higher affinity to Mab A16 than peptide LK[CADPNRFK]GKDL, however, their reactivity was significantly lower compared to the linear ones. Our results clearly show the importance of secondary structure in an antibody binding and demonstrate that even a slight modification of the primary structure dramatically could influence the immune recognition of the synthetic antigens. PMID- 14556906 TI - Temperature dependency and temperature compensation in a model of yeast glycolytic oscillations. AB - Temperature sensitivities and conditions for temperature compensation have been investigated in a model for yeast glycolytic oscillations. The model can quantitatively simulate the experimental observation that the period length of glycolytic oscillations decreases with increasing temperature. Temperature compensation is studied by using control coefficients describing the effect of rate constants on oscillatory frequencies. Temperature compensation of the oscillatory period is observed when the positive contributions to the sum of products between control coefficients and activation energies balance the corresponding sum of the negative contributions. The calculations suggest that by changing the activation energies for one or several of the processes, i.e. by mutations, it could be possible to obtain temperature compensation in the yeast glycolytic oscillator. PMID- 14556908 TI - Negative feedback regulation is responsible for the non-linear modulation of photosynthetic activity in plants and cyanobacteria exposed to a dynamic light environment. AB - Photosynthetic organisms exposed to a dynamic light environment exhibit complex transients of photosynthetic activities that are strongly dependent on the temporal pattern of the incident irradiance. In a harmonically modulated light of intensity I approximately const.+sin(omegat), chlorophyll fluorescence response consists of a steady-state component, a component modulated with the angular frequency of the irradiance omega and several upper harmonic components (2omega, 3omega and higher). Our earlier reverse engineering analysis suggests that the non-linear response can be caused by a negative feedback regulation of photosynthesis. Here, we present experimental evidence that the negative feedback regulation of the energetic coupling between phycobilisome and Photosystem II (PSII) in the cyanobacterium Synechocystis sp. PCC6803 indeed results in the appearance of upper harmonic modes in the chlorophyll fluorescence emission. Dynamic changes in the coupling of the phycobilisome to PSII are not accompanied by corresponding antiparallel changes in the Photosystem I (PSI) excitation, suggesting a regulation limited to PSII. Strong upper harmonic modes were also found in the kinetics of the non-photochemical quenching (NPQ) of chlorophyll fluorescence, of the P700 redox state and of the CO(2) assimilation in tobacco (Nicotiana tabaccum) exposed to harmonically modulated light. They are ascribed to negative feedback regulation of the reactions of the Calvin-Benson cycle limiting the photosynthetic electron transport. We propose that the observed non linear response of photosynthesis may also be relevant in a natural light environment that is modulated, e.g., by ocean waves, moving canopy or by varying cloud cover. Under controlled laboratory conditions, the non-linear photosynthetic response provides a new insight into dynamics of the regulatory processes. PMID- 14556907 TI - Photosystem I trimers from Synechocystis PCC 6803 lacking the PsaF and PsaJ subunits bind an IsiA ring of 17 units. AB - We report a structural characterization by electron microscopy and image analysis of a supramolecular complex consisting of Photosystem I (PSI) and the chlorophyll binding protein IsiA from a mutant of the cyanobacterium Synechocystis PCC 6803 lacking the PsaF and PsaJ subunits. The circular complex consists of a central PSI trimer surrounded by a ring of 17 IsiA units, one less than in the wild-type supercomplex. We conclude that PsaF and PsaJ are not obligatory for the binding of the IsiA ring, and that the size of the PSI complex determines the number of IsiA units in the ring. The resulting number of 17 copies implies that each PSI monomer has a different association to the IsiA ring. PMID- 14556909 TI - Identification of intramembrane hydrogen bonding between 13(1) keto group of bacteriochlorophyll and serine residue alpha27 in the LH2 light-harvesting complex. AB - Intramembrane hydrogen bonding and its effect on the structural integrity of purple bacterial light-harvesting complex 2, LH2, have been assessed in the native membrane environment. A novel hydrogen bond has been identified by Raman resonance spectroscopy between a serine residue of the membrane-spanning region of LH2 alpha-subunit, and the C-13(1) keto carbonyl of bacteriochlorophyll (BChl) B850 bound to the beta-subunit. Replacement of the serine by alanine disrupts this strong hydrogen bond, but this neither alters the strongly red-shifted absorption nor the structural arrangement of the BChls, as judged from circular dichroism. It also decreases only slightly the thermal stability of the mutated LH2 in the native membrane environment. The possibility is discussed that weak H bonding between the C-13(1) keto carbonyl and a methyl hydrogen of the alanine replacing serine(-4) or the imidazole group of the nearby histidine maintains structural integrity in this very stable bacterial light-harvesting complex. A more widespread occurrence of H-bonding to C-13(1) not only in BChl, but also in chlorophyll proteins, is indicated by a theoretical analysis of chlorophyll/polypeptide contacts at <3.5 A in the high-resolution structure of Photosystem I. Nearly half of the 96 chlorophylls have aa residues suitable as hydrogen bond donors to their keto groups. PMID- 14556910 TI - Nitric oxide and peroxynitrite cause irreversible increases in the K(m) for oxygen of mitochondrial cytochrome oxidase: in vitro and in vivo studies. AB - Mitochondrial cytochrome oxidase is competitively and reversibly inhibited by inhibitors that bind to ferrous heme, such as carbon monoxide and nitric oxide. In the case of nitric oxide, nanomolar levels inhibit cytochrome oxidase by competing with oxygen at the enzyme's heme-copper active site. This raises the K(m) for cellular respiration into the physiological range. This effect is readily reversible and may be a physiological control mechanism. Here we show that a number of in vitro and in vivo conditions result in an irreversible increase in the oxygen K(m). These include: treatment of the purified enzyme with peroxynitrite or high (microM) levels of nitric oxide; treatment of the endothelial-derived cell line, b.End5, with NO; activation of astrocytes by cytokines; reperfusion injury in the gerbil brain. Studies of cell respiration that fail to vary the oxygen concentration systematically are therefore likely to significantly underestimate the degree of irreversible damage to cytochrome oxidase. PMID- 14556911 TI - Origin of apparent negative cooperativity of F(1)-ATPase. AB - In order to get insight into the origin of apparent negative cooperativity observed for F(1)-ATPase, we compared ATPase activity and ATPMg binding of mutant subcomplexes of thermophilic F(1)-ATPase, alpha((W463F)3)beta((Y341W)3)gamma and alpha((K175A/T176A/W463F)3)beta((Y341W)3)gamma. For alpha((W463F)3)beta((Y341W)3)gamma, apparent K(m)'s of ATPase kinetics (4.0 and 233 microM) did not agree with apparent K(m)'s deduced from fluorescence quenching of the introduced tryptophan residue (on the order of nM, 0.016 and 13 microM). On the other hand, in case of alpha((K175A/T176A/W463F)3)beta((Y341W)3)gamma, which lacks noncatalytic nucleotide binding sites, the apparent K(m) of ATPase activity (10 microM) roughly agreed with the highest K(m) of fluorescence measurements (27 microM). The results indicate that in case of alpha((W463F)3)beta((Y341W)3)gamma, the activating effect of ATP binding to noncatalytic sites dominates overall ATPase kinetics and the highest apparent K(m) of ATPase activity does not represent the ATP binding to a catalytic site. In case of alpha((K175A/T176A/W463F)3)beta((Y341W)3)gamma, the K(m) of ATPase activity reflects the ATP binding to a catalytic site due to the lack of noncatalytic sites. The Eadie-Hofstee plot of ATPase reaction by alpha((K175A/T176A/W463F)3)beta((Y341W)3)gamma was rather linear compared with that of alpha((W463F)3)beta((Y341W)3)gamma, if not perfectly straight, indicating that the apparent negative cooperativity observed for wild-type F(1)-ATPase is due to the ATP binding to catalytic sites and noncatalytic sites. Thus, the frequently observed K(m)'s of 100-300 microM and 1-30 microM range for wild-type F(1)-ATPase correspond to ATP binding to a noncatalytic site and catalytic site, respectively. PMID- 14556912 TI - Spectroscopic characterization of a semi-stable, charge-separated state in Cu(2+) substituted reaction centers from Rhodobacter sphaeroides. AB - In reaction centers from Rhodobacter sphaeroides exposed to continuous illumination in the presence of an inhibitor of the Q(A)(-) to Q(B) electron transfer, a semi-stable, charge-separated state was formed with halftimes of formation and decay of several minutes. When the non-heme iron was replaced by Cu(2+), the decay of the semi-stable, charge-separated state became much slower than in centers with bound Fe(2+) with about the same rate constant for formation. In Cu(2+)-substituted reaction centers, the semi-stable state was associated with an EPR signal, significantly different from that observed after chemical reduction of the acceptor-side quinone or after illumination at low temperature, but similar to that of an isolated Cu(2+) in the absence of magnetic interaction. The EPR results, obtained with Cu(2+)-substituted reaction centers, suggest that the slow kinetics of formation and decay of the charge-separated, semi-stable state is associated with a structural rearrangement of the acceptor side and the immediate environment of the metal-binding site. PMID- 14556913 TI - Acquisition of photosynthetic capacity by a reaction centre that lacks the Q(A) ubiquinone; possible insights into the evolution of reaction centres? AB - A photosynthetically impaired strain of Rhodobacter sphaeroides containing reaction centres with an alanine to tryptophan mutation at residue 260 of the M polypeptide (AM260W) was incubated under photosynthetic growth conditions. This incubation produced photosynthetically competent strains containing suppressor mutations that changed residue M260 to glycine or cysteine. Spectroscopic analysis demonstrated that the loss of the Q(A) ubiquinone seen in the original AM260W mutant was reversed in the suppressor mutants. In the mutant where Trp M260 was replaced by Cys, the rate of reduction of the Q(A) ubiquinone by the adjacent (H(A)) bacteriopheophytin was reduced by three-fold. The findings of the experiment are discussed in light of the X-ray crystal structures of the wild type and AM260W reaction centres, and the possible implications for the evolution of reaction centres as bioenergetic complexes are considered. PMID- 14556914 TI - Is research on human tissues at a crossroads? PMID- 14556915 TI - Human tissue research: EORTC recommendations on its practical consequences. AB - Improvement in cancer care is possible by applying new treatment modalities, which are emerging from knowledge and discoveries coming from laboratory research. This is possible through international collaboration and the collection of tumour tissues. Creation of a European Organisation for Research and Treatment of Cancer (EORTC) Tumor Bank is a natural step in this direction, by offering tumour sample collection from patients entered in EORTC trials. The aim of such a bank is not only to improve the diagnostic review process, but also to facilitate translational research by allowing clinicians and basic scientists to enter into close collaborations. The EORTC Tissue Research Policy is developed to assure, under the EORTC legal framework, an ethical and scientific review of research projects, guarantee adequate information is given to patients, establish procedures on the use of materials, including legal aspects, and publication policies. Being part of the EU TubaFrost project, the EORTC will provide a common international platform for the use of tissues for research purposes, finding a balance between different laws and assuring scientific progress. PMID- 14556916 TI - EORTC Cancer in the Elderly Task Force guidelines for the use of colony stimulating factors in elderly patients with cancer. AB - Increasing age is not, in itself, a contraindication to cancer chemotherapy. Myelosuppression, however, a common adverse consequence of the administration of many standard-dose chemotherapy regimens to both young and elderly patients with cancer, increases with age. The risk of development of febrile neutropenia may contribute to a reluctance to administer chemotherapy in the elderly patient population. We conducted a detailed literature search (1992-2002) to derive evidence-based conclusions on the value of prophylactic colony-stimulating factor (CSF) administration in elderly patients receiving chemotherapy. Sufficient evidence allows us to affirm that prophylactic granulocyte colony-stimulating factor (G-CSF) reduces the incidence of chemotherapy-induced neutropenia, febrile neutropenia and infections in elderly patients receiving myelotoxic chemotherapy for non-Hodgkin's lymphoma (NHL), small-cell lung cancer (SCLC) or urothelial tumours. Lack of available trial data does not allow similar conclusions to be drawn for other cancers studied, but it is likely that similar benefits would accrue from the use of prophylactic G-CSF. There is insufficient evidence to extend this recommendation to include the use of granulocyte-macrophage colony stimulating factor (GM-CSF). There are insufficient data available to allow the evaluation of the impact of prophylactic CSF on the incidence of toxic deaths in elderly patients with cancer and this is a crucial question for geriatric oncology practice. There is no evidence in elderly patients that the delivery of standard-dose chemotherapy on schedule improves efficacy measures. The data show that febrile neutropenic events are more likely to occur during the first and second cycles of chemotherapy, thus prophylactic measures should be considered early in the course of treatment. Furthermore, since systematic dose reduction can impact on outcome, primary prophylactic use of G-CSF for all elderly patients receiving curative myelotoxic chemotherapy (cyclophosphamide, doxorubicin, vincristine and prednisolone (CHOP) or CHOP-like) is indicated and we suggest a risk-adapted strategy with primary prophylactic G-CSF administration in high-risk patients. Dose intensification, through dose interval reduction, facilitated by prophylactic G-CSF, improved survival in elderly patients with some specific diseases. There is a need for further well-designed studies to identify the elderly patients who will benefit most from prophylactic G-CSF. To achieve this, we strongly urge the design of and participation in further trials. PMID- 14556917 TI - The current position of complementary/alternative medicine in cancer. AB - Complementary/alternative medicine is a tempting option for many cancer patients. Unfortunately, reliable information is not always easy to obtain and some cancer patients may be misled by interested parties. Research in this area is scarce and with numerous therapies we cannot be sure about efficacy or safety. It follows that our current knowledge gaps urgently needs to be filled. PMID- 14556918 TI - Randomised trials comparing chemotherapy regimens for advanced non-small cell lung cancer: biases and evolution over time. AB - We systematically evaluated the evidence from randomised trials comparing various chemotherapy regimens for advanced non-small cell lung cancer. Across 254 eligible trials (42661 patients), no regimens were compared in >6 studies. Twenty six trials (10%) found statistically significant differences in survival between the compared arms. Only five reported the randomisation mode, and four reported adequate allocation concealment; nine performed unaccounted interim analyses. Statistical significance was more common in larger (P=0.003), more recent studies (P=0.031), and trials from countries with only one published eligible study (P=0.008). Increased reported median survival was independently associated with platinum and/or taxane and combination regimens, but also with the year of publication, smaller sample size, and larger representation of non-stage IV patients and patients with a better performance status. The proportion of enrolled patients with a performance status of 2 or worse decreased significantly over time (12.9% per decade, P<0.001). Randomised evidence in this field is fragmented and subject to considerable selection biases. PMID- 14556919 TI - Neuropsychological functioning following systemic treatment in women treated for breast cancer: a review. AB - The aim of this review was to evaluate the effect of treatment and illness related factors on neuropsychological functioning in women treated for breast cancer. Eight studies were identified examining neuropsychological test performance following systemic treatment. Six of the eight studies suggest that neuropsychological functioning may be impaired following treatment. However, there are a number of important methodological issues which limit interpretation of these results. Therefore, it is unclear whether neuropsychological outcome differs according to a range of treatment, biomedical and psychological factors. Larger samples with longitudinal follow-up are required in order to examine the treatment-related factors that best predict cognitive deficits. PMID- 14556920 TI - Frequent methylation of p16INK4A and p14ARF genes implicated in the evolution of chronic myeloid leukaemia from its chronic to accelerated phase. AB - The frequency and mechanism of p16(INK4A) and p14(ARF) gene alterations were studied in cell samples from 30 patients with Philadelphia (Ph) chromosome positive chronic myeloid leukaemia (CML), both at diagnosis and at the onset of the accelerated phase (AP) of the disease. No alterations in the p16(INK4A) or p14(ARF) genes were found in any of the chronic phase (CP) samples. DNA sequencing analyses detected p16(INK4A) or p14(ARF) mutations in 17 AP samples. All mutations were heterozygous without loss of the other allele. Aberrant methylation of the p16(INK4A) or p14(ARF) promoters was found in 14 of 30 AP samples. The most common situation was the simultaneous methylation of both promoters. Our data indicate that p16(INK4A) and p14(ARF) are primary targets for inactivation by promoter methylation in the acceleration of CML. Transcriptional silencing of the p16(INK4A) and p14(ARF) genes may be important in the conversion of CML from the CP to the AP. PMID- 14556921 TI - Gene promoter hypermethylation in oral rinses of leukoplakia patients--a diagnostic and/or prognostic tool? AB - Leukoplakia is the most frequent oral precancerous lesion and shows a variable rate of malignant transformation. We hypothesised that the detection of molecular alterations, like the promoter hypermethylation of DNA, in oral cytological samples from patients at risk of developing primary or recurrent tumours could be a valuable diagnostic and prognostic tool in the management of these lesions. Two groups of patients with differing risks of developing oral squamous cell carcinoma (OSCC) were analysed. DNA was extracted from the oral rinse of each patient. The methylation status of the p16, p14 and MGMT gene promoters was determined using a methylation-specific polymerase chain reaction (MSP). Methylation of p16 and MGMT was observed in 44 and 56% of the oral samples, respectively. Only 12% of the cases showed p14 methylation. DNA hypermethylation was more frequent in patients with previous OSCC. DNA promoter hypermethylation is frequent during early oral carcinogenesis and even more so in the later stages. MSP using oral rinses is a non-invasive and highly sensitive technique which could be used to monitor patients with precancerous and cancerous oral lesions. PMID- 14556922 TI - Efficacy of tamoxifen following anastrozole ('Arimidex') compared with anastrozole following tamoxifen as first-line treatment for advanced breast cancer in postmenopausal women. AB - Anastrozole ('Arimidex') is indicated for the treatment of advanced breast cancer in postmenopausal women. Combined analysis of two international randomised, double-blind trials (n=1021) showed that in patients with hormone receptor positive tumours, first-line treatment with anastrozole significantly prolonged the time to progression (TTP) compared with tamoxifen (median TTP: 10.7 versus 6.4 months, respectively; P=0.022). Second-line tamoxifen following anastrozole, or vice versa, in this trial population was unblinded. The treatments were crossed over and then efficacy was assessed using a questionnaire. Of 511 patients randomised to anastrozole, 137 (26.8%) received second-line tamoxifen. Questionnaire data were available for 119 patients; 58 (48.7%) gained clinical benefit (CB=complete+partial response (CR+PR)+(stable disease (SD) >/=24 weeks)), while 12 (10.1%) had an objective response (OR=CR+PR). Of 510 patients randomised to tamoxifen, 134 (26.3%) received second-line anastrozole. Questionnaire data from 95 patients showed that 54 (56.8%) gained CB and 7 of the patients gaining CB (7.4%) had an OR. Previous studies showed anastrozole is effective after first line tamoxifen. These data show that the sequential administration of first-line anastrozole followed by tamoxifen provides effective use of these drugs in the treatment of postmenopausal women with advanced breast cancer. PMID- 14556923 TI - An open randomised trial of second-line endocrine therapy in advanced breast cancer. comparison of the aromatase inhibitors letrozole and anastrozole. AB - It was previously shown that letrozole (Femara) was significantly more potent than anastrozole (Arimidex) in inhibiting aromatase activity in vitro and in inhibiting total body aromatisation in patients with breast cancer. The objective of this study was to compare letrozole (2.5 mg per day) and anastrozole (1 mg per day) as endocrine therapy in postmenopausal women with advanced breast cancer previously treated with an anti-oestrogen. This randomised, multicentre and multinational open-label phase IIIb/IV study enrolled 713 patients. Treatment was for advanced breast cancer that had progressed either during anti-oestrogen therapy or within 12 months of completing that therapy. Patients had tumours that were either positive for oestrogen and/or progesterone receptors (48%) or of unknown receptor status (52%). The primary efficacy endpoint was time to progression (TTP). Secondary endpoints included objective response, duration of response, rate and duration of overall clinical benefit (responses and long-term stable disease), time to treatment failure, and overall survival, as well as general safety. There was no difference between the treatment arms in TTP; median times were the same for both treatments. Letrozole was significantly superior to anastrozole in the overall response rate (ORR) (19.1% versus 12.3%, P=0.013), including in predefined subgroups (receptor status-unknown, and soft-tissue- and viscera-dominant site of disease). There were no significant differences between the treatment arms in the rate of clinical benefit, median duration of response, duration of clinical benefit, time to treatment failure or overall survival. Both agents were well tolerated and there were no significant differences in safety. These results support previous data documenting the greater aromatase-inhibiting activity of letrozole and indicate that advanced breast cancer is more responsive to letrozole than to anastrozole as second-line endocrine therapy. PMID- 14556924 TI - A phase I study of S-1 combined with weekly cisplatin for metastatic gastric cancer in an outpatient setting. AB - A dose-escalation study was conducted for patients with metastatic gastric cancer to determine the recommended dose of weekly intravenous (i.v.) cisplatin combined with a fixed dose of a new oral dihydropyrimidine dehydrogenase-inhibitory fluoropyrimidine, S-1, on an outpatient basis. Secondary endpoints were to define the toxicity profile and to determine tumour responses. S-1 was fixed at a dose of 70 mg/m(2)/day and was administered for 2 weeks followed by a 1-week rest. Three dose levels of cisplatin (10, 15 and 20 mg/m(2)) were studied. Cisplatin was infused over 30 min on days 1 and 8. 20 patients were enrolled. No dose limiting toxicities (DLTs) were recorded during the administration of cisplatin up to 20 mg/m(2), except for grade 3 diarrhoea and stomatitis in one patient at dose level 3. No grade 4 adverse events occurred. However, grade 2 gastrointestinal adverse reactions, such as nausea and anorexia, were seen in 7 of 13 patients at dose level 3 within the first two treatment cycles. This was determined to be the maximum acceptable level that would not negate the advantages observed with use of an oral drug such as S-1. An objective tumour response was seen at all dose levels, and the overall response rate in the 18 patients evaluated was 61%. A higher response rate of 78% was observed in 9 patients who had received no prior chemotherapy. Oral S-1 with weekly cisplatin is a feasible and promising combination regimen that is appropriate for an outpatient setting. A randomised phase II study comparing this combination with S 1 alone in chemo-nai;ve patients is warranted. PMID- 14556925 TI - Glufosfamide administered using a 1-hour infusion given as first-line treatment for advanced pancreatic cancer. A phase II trial of the EORTC-new drug development group. AB - The activity of glufosfamide (beta-D-glucopyranosyl-N,N'-di-(2-chloroethyl) phosphoric acid diamide) against pancreatic cancer was investigated in a multicentre, phase II clinical study. Chemotherapy-nai;ve patients with advanced or metastatic disease were treated with glufosfamide (5 g/m(2)) using a 1-h intravenous (i.v.) infusion every 3 weeks. Patients were randomised between active-hydration and normal fluids to evaluate the nephroprotective effect of forced diuresis. Patients experiencing >0.4 mg/dl (>35 micromol/l) increase in serum creatinine compared with their baseline value were taken off treatment for safety reasons. The evaluation of response was according to the Response evaluation criteria in solid tumours (RECIST). Blood sampling was performed for pharmacokinetic analyses. 35 patients from 13 institutions were registered over a 13-month period. A total of 114 treatment cycles (median 3, range 1-8) were administered to 34 patients; 18 patients were allocated to the hydration arm. Overall haematological toxicity was mild. Metabolic acidosis occurred in 2 patients treated in the active-hydration arm, grade 3 hypokalaemia was recorded in 5 patients and grade 3 hypophosphataemia in 4 patients. One patient had a grade 4 increase in serum creatinine level, concomitantly to disease progression. Active-hydration did not show a nephroprotective effect and the plasma pharmacokinetics (Pk) of glufosfamide was not significantly influenced by hydration. Two confirmed partial remissions (PR) were reported (response rate 5.9%, 95% Confidence Interval (CI) 0.7-19.7%) and 11 cases obtained disease stabilisation (32.4%). An extra mural review panel confirmed all of the responses. Median overall survival was 5.3 months (95% CI 3.9-7.1) and time to progression (TTP) was 1.4 months (95% CI 1.3-2.7). In conclusion, glufosfamide administered using a 1-h infusion every 3 weeks has a modest activity in advanced pancreatic adenocarcinoma. Haematological toxicity is particularly mild, but regular monitoring of renal function is recommended. PMID- 14556926 TI - Langerhans cell histiocytosis in adults. Report from the International Registry of the Histiocyte Society. AB - Langerhans cell histiocytosis (LCH), characterised by the infiltration of one or more organs by large mononuclear cells, can develop in persons of any age. Although the features of this disease are well described in children, they remain poorly defined in adults. From January 2000 to June 2001, 274 adults from 13 countries, with biopsy-proven adult LCH, were registered with the International Histiocyte Society Registry. Information was collected about clinical presentation, family history, associated conditions, cigarette smoking and treatment, to assist in future management decisions in patients aged 18 years and older. There were slightly more males than females (143:126), and the mean ages at the onset and diagnosis of disease were 33 years (standard deviation (S.D.) 15 years) and 35 years (S.D. 14 years), respectively. 2 patients had consanguineous parents, and 1 had a family history of LCH; 129 reported smoking (47.1%); 17 (6.2%) had been diagnosed with different types of cancer. Single-system LCH, found in 86 patients (31.4%), included isolated pulmonary involvement in 44 cases; 188 patients (68.6%) had multisystem disease; 81 (29.6%) had diabetes insipidus. Initial treatment consisted of vinblastine administered with or without steroids, to 82 patients (29.9%), including 9 who had received it with etoposide, which was the sole agent given to 19 patients. 236 patients were considered evaluable for survival. At a median follow-up of 28 months from diagnosis, 15 patients (6.4%) had died (death rate, 1.5/100 person years, 95% Confidence Interval (95% CI) 0.9-2.4). The probability of survival at 5 years postdiagnosis was 92.3% (95% CI 85.6-95.9) overall, 100% for patients with single system disease (n=37), 87.8% (95% CI 54.9-97.2) for isolated pulmonary disease (n=34), and 91.7% (95% CI 83.6-95.9) for multisystem disease (n=163). Survival did not differ significantly among patients with multisystem disease, with or without liver or lung involvement) 5-year survival 93.6% (95% CI 84.7-97.4) versus 87.5% (95% CI 65.5-95.9), respectively; P value 0.1). LCH in adults is most often a multisystem disease with the highest mortality seen in patients with isolated pulmonary involvement. It should be included in the differential diagnosis of disseminated or localised disease of the bone, skin and mucosa, as well as the lung and the endocrine and central nervous system, regardless of the age of the patient. A prospective international therapeutic study is warranted. PMID- 14556927 TI - Nasopharyngeal carcinoma in childhood and adolescence: analysis of a series of 32 patients treated with combined chemotherapy and radiotherapy. AB - Standard therapy for nasopharyngeal carcinoma (NPC) in children has generally followed the guidelines established for adults. We report here, the treatment outcomes in 32 children and adolescents with NPC and we discuss treatment approaches. Between 1993 and 1997, 32 NPC patients aged 38.5 degrees C, and had not been vaccinated with BCG or vaccinia. The OR was 0.29 (CI: 0.15-0.57) in those who had had a severe infectious disease and were vaccinated with either BCG or vaccinia and 0.33 (CI: 0.17-0.65) for those with 1 or more severe infectious diseases and who had received both vaccinations. We conclude that both vaccinations as well as previous episodes of having a severe infectious disease induced the same protective mechanism with regards to the risk of melanoma. Because of a 'masking effect' by the vaccinia vaccination, the protective effect of the BCG vaccination and of certain infectious diseases against cancer has remained undetected. The vaccinations contributed more to the protection of the population than a previous episode of having an infectious disease. In view of the termination of vaccinations with vaccinia in all countries and of BCG in many of them, these findings call for a re-evaluation of vaccination strategies. PMID- 14556931 TI - Radiation dose as a risk factor for malignant melanoma following childhood cancer. AB - The aim of this study was to determine therapy-related risk factors for the development of melanoma after childhood cancer. Among 4401 3-year survivors of a childhood cancer in eight French and British centres and 25120 patients younger than 20 years old at first malignant neoplasm (FMN) extracted from the Nordic Cancer Registries, 16 patients developed a melanoma as a second malignant neoplasm (SMN). A cohort study of the French and British cohorts was performed. In a nested case-control study, the 16 patients who developed a melanoma as a SMN (cases) were matched with 3-5 controls in their respective cohort according to gender, age at the first cancer, the calendar year of occurrence of the first cancer and follow-up. Radiotherapy appeared to increase the risk of melanoma for local doses >15 Gy, Odds Ratio (OR)=13 (95% Confidence Interval (CI): 0.94-174). Regarding chemotherapy, we observed an increased OR for both alkylating agents and spindle inhibitors, OR=2.7 (95% CI: 0.5-14). Children treated for a gonadal tumour as a FMN were found to be at a higher risk of melanoma, OR=8.7 (95% CI: 0.9-86). The adjusted OR for the local radiation dose was 1.07 (95% CI: 1.00 1.15). In conclusion, radiotherapy may contribute to an increased risk of melanoma as a SMN, but only at very high doses of low linear energy transfer radiation. Common genetic origins between gonadal tumours and malignant melanomas are likely. PMID- 14556932 TI - Superior antitumour activity of S-1 in tumours with a high dihydropyrimidine dehydrogenase activity. AB - To elucidate the mechanism of the enhanced antitumour activity of S-1 (1 M tegafur, 0.4 M 5-chloro-2, 4-dihydroxypyridine, and 1 M potassium oxonate) in terms of the phosphorylation and degradation pathways of 5-fluorouracil (5-FU) metabolism, we investigated tumoral thymidylate synthase (TS) content, dihydropyrimidine dehydrogenase (DPD) activity, the TS inhibition rate (TS-IR), and 5-FU incorporated into RNA (F-RNA) in four human gastric cancer xenografts (MKN-28, MKN-74, GCIY and GT3TKB) and compared the results obtained with S-1 with those obtained with 5-FU and UFT (1 M tegafur, 4 M uracil). 5-FU was administered intraperitoneally (i.p.) to mice at a dose of 50 mg/kg, three times, on days 0, 4 and 8. S-1 and UFT were administered orally at doses of 10 and 24 mg/kg, respectively, once a day, for 9 consecutive days. Antitumour activity was evaluated as the maximum inhibition of tumour growth in each animal. S-1 showed a better antitumour activity than 5-FU and UFT in tumours with a high DPD activity (GCIY and GT3TKB). There were inverse correlations between the antitumour activity and both TS content and DPD activity in the 5-FU and UFT groups. However, no such correlations were observed in the S-1 group. In GCIY and GT3TKB xenografts, TS-IR was significantly higher in the S-1 group than in the 5-FU or UFT groups. In GT3TKB xenografts, the F-RNA level was significantly higher in the S-1 group than in the 5-FU or UFT groups. The superior cytotoxicity of S-1 appears to be attributable to both an increased inhibition of DNA synthesis and an enhanced blockade of RNA function against tumours with a high DPD activity. PMID- 14556933 TI - Acquisition of anoikis resistance in human osteosarcoma cells. AB - Under normal circumstances, adhered cells die of anoikis when detached from their extracellular matrix (ECM). Resistance to anoikis has been implicated in the progression of many human malignancies by affording an increased survival time in the absence of matrix attachment, facilitating the migration and eventual colonisation of distant sites. In this study, an anoikis-resistant variant of the human osteosarcoma cell line, SAOS-2 (SAOSar), was generated by sequential cycles of culturing under adhered and suspended conditions. It was also shown that although parental SAOS (SAOSp) cells are a heterogeneous population with varying levels of sensitivity to anoikis, the establishment of anoikis-resistant clones was not necessarily the result of mere selection of a previously resistant subpopulation. Anoikis-resistant cells were also derived from anoikis-sensitive SAOS clones by exposure to anoikis-inducing culture conditions. This suggests that lack of the normal signalling generated by attachment to the ECM could represent a driving force towards anoikis resistance. Resistance to anoikis could not be attributed to a general defect in the apoptotic pathway since apoptosis in both sensitive and resistant populations was induced after treatment with staurosporine, cycloheximide and hydrogen peroxide. This suggests that the apoptotic machinery is intact in both anoikis-sensitive and -resistant SAOS cells and that the death signal in anoikis-sensitive cells is generated by the lack of attachment, most probably by unligated integrins. Anoikis-resistant cells have circumvented this death signal and remain viable despite suspended conditions. PMID- 14556934 TI - Antitumour activity of the silybin-phosphatidylcholine complex, IdB 1016, against human ovarian cancer. AB - This study aimed to assess, in an in vivo experimental model, the growth inhibitory effects of IdB 1016 (Silipide, a complex of silybin/phosphatidylcholine) when used as a single agent against human ovarian cancer. We also wanted to investigate the mechanism of the antiangiogenic action by assessing Vascular Endothelial Growth Factor (VEGF) levels and by using macroarray technology to evaluate the regulation of a panel of genes involved in angiogenesis. We also aimed to establish the plasma and tumour bioavailability of silybin after repeated administration of IdB 1016. Female nude mice bearing human ovarian cancer xenografts (A2780) received 450 mg/kg/day IdB 1016 daily by oral gavage until the end of the study. At sacrifice, blood and tumour specimens were collected and subsequently processed for the determination of silybin levels, VEGF levels or a gene expression profile. IdB 1016 was significantly active in inhibiting ovarian tumour growth. Treatment with 450 mg/kg/day for a total of 20 administrations produced a tumour weight inhibition (TWI%) of 78% and a Log10 Cell Kill (LCK) of 1.1. Free silybin levels were found to be 7.0+/-5.3 microg/ml and 183.5+/-85.9 ng/g tissue (mean+/-standard deviation (S.D.)) in the plasma and tumour samples, respectively. No significant differences were found in the concentration of human VEGF in xenografts from control and IdB 1016-treated mice. The array analysis suggested the downregulation of the VEGR receptor 3 and the upregulation of angiopoietin-2 as potential mechanisms for the antiangiogenic activity. In conclusion, these findings suggest IdB 1016 is a good candidate, with a relevant clinical potential, for use in the management of recurrent ovarian cancer. A phase II, non-randomised clinical study is now ongoing in our Institute aimed at evaluating the efficacy of daily administrations of IdB 1016 in the serological recurrence of ovarian cancer. PMID- 14556935 TI - Distribution of a GABAB-like receptor protein in the rat central nervous system. AB - Using a homology-based bioinformatics approach we have identified the human and rodent orthologues of a novel putative seven transmembrane G protein coupled receptor, termed GABA(BL). The amino acid sequence homology of these cDNAs compared to GABA(B1) and GABA(B2) led us to postulate that GABA(BL) may be a putative novel GABA(B) receptor subunit. We have developed a rabbit polyclonal antisera specific to the GABA(BL) protein and assessed the distribution of GABA(BL) in the rat CNS by immunohistochemistry. Protein expression was particularly dense in regions previously shown to contain known GABA(B) receptor subunits. Dense immunoreactivity was observed in the cortex, major subfields of the hippocampus and the dentate gyrus. GABA(BL) labelling was very conspicuous in the cerebellum, both in the granule cell layer and in Purkinje cells, and was also observed in the substantia gelatinosa and ventral horn motor neurons of the spinal cord. GABA(BL) immunoreactivity was also noted in a subset of parvalbumin positive hippocampal interneurons. Our data suggest a widespread distribution of GABA(BL) throughout the rat CNS. PMID- 14556936 TI - Specific oligobodies against human brain acetylcholinesterase. AB - In order to develop the specific oligobodies against human brain acetylcholinesterase (AChE) and distinguishes between human erythrocyte and brain AChEs, we applied the strategy of 'target switching' to obtain the specific polyclonal and monoclonal oligobodies. The specificity between human brain AChE and other ChEs was identified by Western blotting, dot blotting and enzyme protein binding assay (EPBA). The results showed that the oligobodies against the human brain AChE specifically immunoreacted with the human brain AChE and Torpedo AChE, not showing significant binding to AChE from human erythrocyte and butyrylcholinesterase (BChE) from human serum. PMID- 14556937 TI - Rhythmic and discrete elements in multi-joint coordination. AB - Everyday actions invariably consist of a combination of discrete and rhythmic elements within or across joints. The study investigated constraints arising from the co-occurrence of the two actions in a two-joint task and how endpoint trajectories are shaped due to these action elements at the joint level. The task consisted of an elbow oscillation in the plane that was to be merged with a fast discrete adduction or abduction in the shoulder triggered by an auditory signal. The task was performed with and without explicit instruction about the joint involvement. Two hypotheses were tested: (1) kinematic constraints for the coupling of discrete and rhythmic elements arise at the neuro-muscular level, such that EMG bursts of the discrete and rhythmic movement have a tendency to synchronize. This effect was documented previously in a comparable single-joint task. (2) The merging of the two elements is constrained by intersegmental torques such that initiation and performance of the discrete movement utilizes interaction torques. This hypothesis rests on the assumption that the CNS has an internal model of the limb dynamics and exploits passive torques. Key results support hypothesis 1: (i) the discrete action's initiation at the shoulder was constrained to a preferred phase of the ongoing elbow oscillation. (ii) The rhythmic elbow movement showed a systematic phase advance during the discrete shoulder shift, similar to those reported for the single-joint variant of the task. Reaction times of the discrete movement were longer and peak velocities slower than reported for isolated discrete movements, due to the simultaneous presence of the oscillation. (iii) Interaction torques acting from the elbow onto the shoulder joint were not selectively exploited for the acceleration of the discrete shoulder movement. Indirectly however, hypothesis 2 also found support: torques at the elbow generated compensatory muscle activity in the shoulder that stabilized the stationary joint. It was this rhythmic activity that posed the direct constraints on the initiation of the discrete movement. PMID- 14556938 TI - Upregulation of oligodendrocyte progenitor cells associated with restoration of mature oligodendrocytes and myelination in peri-infarct area in the rat brain. AB - This study examines the alteration of oligodendrocyte progenitor cells (OPCs), mature oligodendrocytes (OLGs) and myelination after focal ischemia in the rat brain. Adult male Sprague-Dawley rats were subjected to 90-min occlusion of the middle cerebral artery, followed by reperfusion time of up to 2 weeks. The infarct core showed a rapid and progressive decrease in the number of OPCs, OLGs, as well as the myelin density after 48 h of recirculation. The peri-infarct area exhibited a moderate reduction in the number of OLGs and the myelin density with a slight increase in the number of OPCs at 48 h of recirculation. Subsequently, a steady increase in the number of OPCs and a gradual recovery of the number of OLGs were noted in the peri-infarct area, which were accompanied by a gradual restoration of the myelin density, resulting in almost complete recovery of myelin density at 2 weeks of recirculation. OPCs in the peri-infarct area showed characteristic morphological changes such as mitotic figures, monopolar or bipolar shapes, and hypertrophied cell bodies and processes, all indicating active cell proliferation and migration. These findings suggest that the upregulation of OPCs may contribute to replenishment of OLGs and resultant remyelination in the peri-infarct area after ischemic insult. PMID- 14556939 TI - Possible mechanisms underlying the protective effects of SY-21, an extract of a traditional Chinese herb, on transient brain ischemia/reperfusion-induced neuronal death in rat hippocampus. AB - We examined the neuroprotective actions of SY-21, a potent ingredient of a traditional Chinese herb, histologically. Transient brain ischemia (15 min) was induced by four-vessel occlusion in Sprague-Dawley rats. Administration of SY-21 20 min before or 6 h after brain ischemia significantly increased the number of surviving hippocampal CA1 pyramidal cells in rats subjected to transient brain ischemia followed by 5 days of reperfusion. Neuronal cell death resulting from ischemic events is associated with abnormal activation of the NMDA receptors. Tyrosine phosphorylation of the NMDA receptor subunit NR2A by Src or Fyn has been implicated in the up-regulation of NMDA receptor activity. In order to investigate the possible mechanism of the neuroprotective action of SY-21, we examined the effects of SY-21 on the ischemia/reperfusion-induced increases in tyrosine phosphorylation of the NMDA receptor subunit 2A (NR2A) and on the interactions involving NR2A, PSD-95 and Src/Fyn. We found that the increase in the tyrosine phosphorylation of NR2A induced by brain ischemia/reperfusion is suppressed by SY-21 administered 15 min before, or instantly after, brain ischemia. Also, SY-21 attenuated the increased interactions involving NR2A, PSD 95, Fyn and Src. These results demonstrate that SY-21 has a prominent neuroprotective action against brain ischemic insult, and the mechanism may involve the regulation of the tyrosine phosphorylation of NR2A by changing the above interactions. PMID- 14556940 TI - Apparent opposite effects of tetrabenazine and reserpine on the toxic effects of 1-methyl-4-phenylpyridinium or 6-hydroxydopamine on nigro-striatal dopaminergic neurons. AB - It is well documented that VMAT2 protects nigrostriatal DA neurons against MPP(+) by sequestering it inside vesicles away from its mitochondrial site of neurotoxic action. However, the implication of the VMAT2 in the mechanism of action exerted by 6-OHDA has received little attention. Therefore, the aim of the present study was to determine whether the vesicular sequestration of 6-OHDA would protect dopaminergic neurons from its toxicity similarly to what is observed with MPP(+). We injected mice with 6-OHDA 90 min after TBZ treatment. Since, unexpectedly, TBZ pretreatment prevented 6-OHDA neurotoxicity, we performed a similar experience replacing 6-OHDA with MPP(+) in order to check our experimental protocol. TBZ pretreatment similarly prevented MPP(+) neurotoxicity. This discrepancy with what is commonly describe in the literature, led us to use reserpine. Indeed, the long lasting VMAT2 inhibition induced by reserpine allowed us to inject neurotoxins while mice no longer presented hypothermia. Contrary to TBZ pretreatment, reserpine pretreatment potentiated both 6-OHDA and MPP(+) toxicity on dopaminergic neurons. Hypothermia elicited by TBZ appeared to be responsible, at least in part, for the neuroprotective effect observed. To verify this hypothesis, we investigated the influence of hypothermia on the toxic activity of both neurotoxins. A hypothermia similar to that induced by TBZ was obtained by a forced swimming test of putting mice into cool water (23 degrees C). The hypothermia prevented both 6-OHDA and MPP(+)-induced neurotoxicity. We finally reported that VMAT2 inhibition potentiates both MPP(+) and 6-OHDA neurotoxicity. PMID- 14556941 TI - Riluzole suppresses experimental autoimmune encephalomyelitis: implications for the treatment of multiple sclerosis. AB - Recent studies suggest that glutamate neurotoxicity is involved in the pathogenesis of multiple sclerosis (MS), and that treatment with glutamate receptor (AMPA/kainate) antagonists inhibits experimental autoimmune encephalomyelitis (EAE), the conventional model of MS. Therefore, we examined whether riluzole, an inhibitor of glutamate transmission, affects the pathogenesis and clinical features of MS-like disease in myelin oligodendrocyte glycoprotein (MOG)-induced EAE in mice. Here we report that riluzole (10 mg/kgx2/day, i.p.), administered before and even after the appearance of clinical symptoms, dramatically reduced the clinical severity of MOG-induced EAE, while all the MOG-immunized control mice developed significant clinical manifestations. Moreover, the riluzole-treated mice demonstrated only mild focal inflammation, and less demyelination, compared to MOG-treated mice, using histological methods. Furthermore, riluzole markedly reduced axonal disruption, as assessed by Bielshowesky's silver staining and by antibodies against non-phosphorylated neurofilaments (SMI-32). No difference was detected in the immune system potency, as T-cell proliferative responses to MOG were similar in both groups. In conclusion, our study demonstrates, for the first time, that riluzole can reduce inflammation, demyelination and axonal damage in the CNS and attenuate the clinical severity of MOG-induced EAE. These results suggest that riluzole, a drug used in amyotrophic lateral sclerosis (ALS), might be beneficial for the treatment of MS. PMID- 14556942 TI - 3,4-Dihydroxyphenylacetaldehyde is the toxic dopamine metabolite in vivo: implications for Parkinson's disease pathogenesis. AB - In Parkinson's disease (PD), there is a highly selective loss of dopamine (DA) neurons in the substantia nigra (SN) greater than in the ventral tegmental area (VTA). The simplest explanation for selective DA neuron loss in PD is that DA is toxic and, because only DA neurons contain significant amounts of DA, this highly localized synthesis of DOPAL accounts for selective vulnerability of DA neurons. However, the large concentrations of DA required to produce in vivo toxicity cast doubt on its role in PD pathogenesis. Alpha-synuclein (alpha-syn) is the major component of the Lewy body, the pathological marker of PD, and is genetically linked to the disease. Recent studies indicate that alpha-syn neurotoxicity is mediated by a free radical generating metabolite of DA. Here we test the hypothesis that 3,4-dihydroxyphenylacetaldehyde (DOPAL), the monamine oxidase metabolite of DA, mediates DA toxicity in vivo. We injected DOPAL, DA and its oxidative, reduced and methylated metabolites into rat SN and VTA. Five days post surgery, the injection sites were evaluated in Nissl preparations and with tyrosine hydroxylase (for DA neurons), neuronal nuclear antigen (for neurons) and glial fibrillary acidic protein (for astrocytes) immunoreactivities. Lesion size in SN vs. VTA was compared using morphometry. DOPAL at concentrations as low as 100 ng was toxic to DA SN neurons>DA VTA neurons>glia. Neither DA nor its other metabolites showed evidence of neurotoxicity at fivefold higher doses. However, 20 microg of DA produced lesions in the SN and VTA. We conclude that DOPAL is the toxic DA metabolite in vivo. Implications for a unified hypothesis for PD pathogenesis are discussed. PMID- 14556943 TI - P2X3-immunoreactive primary sensory neurons innervating lumbar intervertebral disc in rats. AB - The P2X(3) receptor is normally localized in a sub-population of small-diameter dorsal root ganglion (DRG) neurons, and is thought to be related to pain perception. The aim of this study in rats was to examine P2X(3)-immunoreactivity in DRG neurons innervating the lumbar disc and in DRG neurons innervating cutaneous tissues. Fluoro-Gold was applied to the L5-L6 disc, the plantar skin of the hind paw (L4-L5 dermatomes), and the back skin (L1-L2 dermatomes). It has been reported that the L5-L6 disc is innervated by T13-L5 DRG neurons. We performed immunostaining using antibodies against the P2X(3) receptor of T13-L5 DRGs to examine the L5-L6 disc, L4 and L5 DRGs to examine plantar skin and L1 and L2 DRGs to examine back skin. The P2X(3)-immunoreactivity was detected in 22.0 and 22.8% of neurons, labeled by Fluoro-Gold applied to plantar and back skin, respectively. However, P2X(3)-immunoreactivity was detected in only 4.0% of the neurons projecting to the L5-L6 disc. The proportion of P2X(3)-immunoreactive neurons was significantly larger in the DRG neurons innervating the plantar or the back skin, than in the DRG neurons innervating the lumbar disc. These results suggest that the P2X(3) receptors are abundant in DRG neurons innervating cutaneous tissues, but not in neurons innervating the lumbar disc. It is likely therefore that the P2X(3) receptor is less related to the mechanism of discogenic pain, than to cutaneous tissue pain. PMID- 14556945 TI - Hypoxic induction of vascular endothelial growth factor is selectively impaired in mice carrying the mutant SOD1 gene. AB - Localization and hypoxic induction of vascular endothelial growth factor (VEGF) was examined in the spinal cord of transgenic mice carrying a mutation in the superoxide dismutase 1 gene. Immunohistochemical and immunofluorescent study demonstrated that VEGF is mainly expressed in motor neurons before and after hypoxia. Baseline expression of VEGF was higher in transgenic (Tg) mice than in wild-type (Wt) littermates. However, VEGF was hardly induced after hypoxia in Tg mice, whereas Wt mice showed an approximate nine-fold increase. Impaired VEGF induction was evident in Tg mice at 12 weeks of age, when they were still presymptomatic. In contrast, baseline and hypoxic expression of brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor did not differ between Tg and Wt mice. Thus, the present study demonstrates that hypoxic induction of VEGF in Tg mice is selectively impaired from a very early stage, suggesting profound involvement in the pathogenesis of motor neuron degeneration in this animal model of amyotrophic lateral sclerosis. PMID- 14556944 TI - Losses of NG2 and NeuN immunoreactivity but not astrocytic markers during early reperfusion following severe focal cerebral ischemia. AB - The ability of glia to recover essential functions following a period of focal cerebral ischemia is likely to be one important factor influencing the severity of tissue damage that subsequently develops. In this study, we have compared changes in immunoreactivity of markers specific for astrocytes, NG2-positive glia and neurons in tissue subregions during early reperfusion following 3 h of middle cerebral artery occlusion to provide insights into possible differential susceptibility of these cell populations. Under the conditions used, infarction ultimately encompasses most of the perfusion territory of the occluded artery. Nonetheless, alterations in immunoreactivity during the first 3 h of recirculation were restricted to brain regions that had been subjected to severe ischemia. In the striatum, cellular immunoreactivity for NG2 and neuronal markers, NeuN and microtubule-associated protein 2, was greatly reduced by 1 h of reperfusion and declined further at 3 h. NG2 labeling of blood vessels in the striatum appeared post-ischemically, mimicking expression of this protein during development. Less severe changes were seen in the neuronal markers in overlying cerebral cortex. In contrast to the losses of other cellular proteins, immunoreactivity for the astrocytic marker, glial fibrillary acidic protein, was preserved in all tissue that had been subjected to severe ischemia and labeling of another astrocytic protein, glutamine synthetase, was increased by 3 h of reperfusion. These findings provide the first evidence of marked sensitivity of NG2-immunoreactivity to severe ischemia and suggest a greater initial resistance of astrocytes compared with neurons and NG2-positive glia to ischemia-reperfusion damage. PMID- 14556946 TI - Tremor in the human hand following peripheral nerve transection and reinnervation. AB - Slow movements and position holding by the digits are both characterised by 8-10 Hz tremor which appears to be centrally generated. Denervation and subsequent reinnervation lead to significant alterations in peripheral connectivity and reflex organisation. We have tested the hypothesis that 8-10 Hz tremor is present in the digits of subjects following a complete nerve lesion. The frequency content of abduction and adduction movements was recorded in 12 index fingers and nine little fingers reinnervated subsequent to a complete ulnar nerve transection. An optical position laser transducer was used to measure digital movements, minimising mechanical interference to the system. Concurrently, surface electromyograms (EMG) were also recorded from first dorsal interosseus muscles (1DI) and abductor digiti minimi brevis (ADMB) muscles for index and little fingers, respectively. The maximal voluntary contraction (MVC) of the reinnervated muscles varied from 5.9% to 100% of those of the unimpaired, contralateral hands. The subjects performed abduction-adduction movements of the index and little fingers and a position holding task. Significant peaks in PSD curves of acceleration and rectified integrated EMG traces were identified. Tremor in the 8-10 Hz range was evident in both the acceleration and EMG signals for the majority of digits during both the slow movement and position holding tasks. These findings demonstrate the robust nature of these 8-10 Hz oscillations, even following the significant changes in peripheral connectivity of muscle and nerve resulting from nerve transection and reinnervation. PMID- 14556948 TI - Arousal in Drosophila. AB - Arousal can be described as an endogenously generated or exogenously induced change in behavioral responsiveness. Changes in levels of arousal, such as occur during sleep or attention, most likely accomplish adaptive functions common to most animals. Recent evidence demonstrating changing arousal states in Drosophila melanogaster complements other behavioral research in this model organism. Herein we review the methodology related to the study of circadian rhythms, sleep and anesthesia where arousal, or lack of it, plays an essential role. We end this review by discussing a new method that allows for the first time to correlate changes in brain electrophysiology to changes in behavioral arousal in the fruit fly. PMID- 14556947 TI - Prenatal stress in rats predicts immobility behavior in the forced swim test. Effects of a chronic treatment with tianeptine. AB - Prenatally-stressed (PS) rats are characterized by a general impairment of the hypothalamo-pituitary-adrenal (HPA) axis and sleep disturbances indicating that this model has face validity with some clinical features observed in a subpopulation of depressed patients. The prolonged corticosterone secretion shown by PS rats in response to stress was positively correlated with an increased immobility behavior in the forced swim test. To investigate the predictive validity of this model, a separate group of animals was chronically treated with the antidepressant tianeptine (10 mg/kg i.p. for 21 days). Such chronic treatment reduced in PS rats immobility time in the forced swim test. These findings suggest that the PS rat is an interesting animal model for the evaluation of antidepressant treatment. PMID- 14556949 TI - Locomotor activity: a complex behavioural trait to unravel. AB - Locomotor activity in Drosophila, as in other organisms, is an important trait since it is at the basis of almost all behaviours. Indeed, the locomotor centre is implicated in all complex behaviours consisting of a change in the position of the animal with respect to its environment. Despite its importance, locomotor activity itself has received sparse attention for the following two reasons: first, until recently, the study of locomotor activity has lacked a well automated and standardised paradigm which is necessary for a detailed description. Second, locomotor activity is complicated by many factors (genetic, feeding, temperature), and as such is rather difficult to study. With recent technological developments, locomotor activity is now more accessible to automated paradigms. These have permitted us to reveal that locomotor activity is a very complex and rich behaviour that follows strict rules, harbours an organised (fractal-like) structure, and consequently might adhere to highly organised neurophysiological processes. Undoubtedly, locomotor activity has now reached a scientific maturity that allows it to be studied with the panoply of neuroethological approaches, in particular genetic, to unravel its mechanisms and neural circuitry. Consequently, we propose that locomotor activity can now represent a relevant biomarker to study various model diseases such as addiction, Parkinson, Alzheimer, Huntington, and diabetes. PMID- 14556950 TI - Circadian rhythms of locomotor activity in Drosophila. AB - Drosophila is by far the most advanced model to understand the complex biochemical interactions upon which circadian clocks rely. Most of the genes that have been characterized so far were isolated through genetic screens using the locomotor activity rhythms of the adults as a circadian output. In addition, new techniques are available to deregulate gene expression in specific cells, allowing to analyze the growing number of developmental genes that also play a role as clock genes. However, one of the major challenges in circadian biology remains to properly interpret complex behavioral data and use them to fuel molecular models. This review tries to describe the problems that clockwatchers have to face when using Drosophila activity rhythms to understand the multiple facets of circadian function. PMID- 14556951 TI - Experimental studies of adult Drosophila chemosensory behaviour. AB - Drosophila melanogaster is a powerful animal model to study the processes underlying behavioural responses to chemical cues. This paper provides a review of the important literature to present recent advances in our understanding of how gustatory and olfactory stimuli are perceived. An overview is given of the experimental procedures currently used to characterize the fly chemosensory behaviour. Since this species provides extremely useful genetic tools, a focus is made on those allowing to manipulate behaviour, and hence to understand its molecular and cellular bases. Such tools include single-gene mutants and the Gal4/UAS system. They can be combined with studies of the natural polymorphism of behavioural responses. Recent data obtained with these various approaches unravel some important aspects of taste and olfaction. These appear as rather complex processes, as revealed by results showing dose-dependence, plasticity and sexual dimorphism. Taken together, these results and the available tools open interesting perspectives for the years to come, in our attempts to make the link between genes and behaviour. PMID- 14556953 TI - The voyeurs' guide to Drosophila melanogaster courtship. AB - Drosophila melanogaster has a long and distinguished history as a model organism in studies of sex-specific behaviour. Courtship is relevant to a wide variety of areas of biological research, from investigations into the evolution of sex specific behaviours, to studies of the molecular mechanisms underlying the perception and processing of sex-specific information, and the identification of genes that enable a fly to behave in a sex-specific fashion. To address any of these issues it is essential that courtship behaviour is investigated in a robust, reproducible and reliable manner. In this review I consider many of the problems that one might encounter in a study of Drosophila courtship, and how such issues may be addressed. PMID- 14556954 TI - Associative learning and memory in Drosophila: beyond olfactory conditioning. AB - The associative learning abilities of the fruit fly, Drosophila melanogaster, have been demonstrated in both classical and operant conditioning paradigms. Efforts to identify the neural pathways and cellular mechanisms of learning have focused largely on olfactory classical conditioning. Results derived from various genetic and molecular manipulations provide considerable evidence that this form of associative learning depends critically on neural activity and cAMP signaling in brain neuropil structures called mushroom bodies. Three other behavioral learning paradigms in Drosophila serve as the main subject of this review. These are (1) visual and motor learning of flies tethered in a flight simulator, (2) a form of spatial learning that is independent of visual and olfactory cues, and (3) experience-dependent changes in male courtship behavior. The present evidence suggests that at least some of these modes of learning are independent of mushroom bodies. Applying targeted genetic manipulations to these behavioral paradigms should allow for a more comprehensive understanding of neural mechanisms responsible for diverse forms of associative learning and memory. PMID- 14556955 TI - "Drosophila behaviour: concepts and tools" from arousal to memory in fruitflies. PMID- 14556957 TI - Chronic Helicobacter pylori infection results in gastric hypoacidity and hypergastrinemia in wild-type mice but vagally induced hypersecretion in gastrin deficient mice. AB - Helicobacter pylori infection is a causal factor of gastric cancer (which is associated with low gastric acid secretion) or duodenal ulcer (high acid secretion). Parietal cells and ECL cells in the stomach are controlled by gastrin, which plays a crucial role in the regulation of acid secretion. The present study was undertaken to identify a possible role of gastrin in determining the different responses of the parietal cells and ECL cells to chronic H. pylori infection. Wild-type (C57BL/6J) gastrin(+/+) mice and gastrin( /-) knockout mice, generated through targeted gene disruption and backcrossed eight times to C57BL/6J, were infected with H. pylori for 9 months. The acid output was measured 4 h after pylorus ligation (known to cause vagal excitation). The gastric mucosa was examined by immunocytochemistry with antisera to alpha subunit of H+/K(+)-ATPase for the parietal cells, and to histamine and vesicle monoamine transporter-2 for the ECL cells, and by quantitative electron microscopy. In infected gastrin(+/+) mice, the acid output and the percentage of secreting parietal cells (freely fed state) were 20-30% of the values in uninfected controls, while the density and ultrastructure of parietal cells were normal. The infected mice had hypergastrinemia and displayed hypertrophy and hyperplasia of ECL cells. Although uninfected gastrin(-/-) mice had lower the acid output than uninfected gastrin(+/+) mice, there was a higher acid output (approximately 3 times) in infected gastrin(-/-) mice than their uninfected homologues. The numbers of parietal cells and ECL cells remained unchanged in infected gastrin(-/-) mice. In conclusion, chronic H. pylori infection results to impaired parietal-cell function (acid hyposecretion), hypergastrinemia and hyperplasia of ECL cells in wild-type mice but leads to vagally induced hypersecretion in gastrin-deficient mice. PMID- 14556956 TI - Antinociceptive role of oxytocin in the nucleus raphe magnus of rats, an involvement of mu-opioid receptor. AB - Recent studies showed that oxytocin plays an important role in nociceptive modulation in the central nervous system. The present study was undertaken to investigate the role of oxytocin in antinociception in the nucleus raphe magnus (NRM) of rats and the possible interaction between oxytocin and the opioid systems. Intra-NRM injection of oxytocin induced dose-dependent increases in hindpaw withdrawal latencies (HWLs) to noxious thermal and mechanical stimulation in rats. The antinociceptive effect of oxytocin was significantly attenuated by subsequent intra-NRM injection of the oxytocin antagonist 1-deamino-2-D-Tyr-(Oet) 4-Thr-8-Orn-oxytocin. Intra-NRM injection of naloxone dose-dependently antagonized the increased HWLs induced by preceding intra-NRM injection of oxytocin, indicating an involvement of opioid receptors in oxytocin-induced antinociception in the NRM of rats. Furthermore, the antinociceptive effect of oxytocin was dose-dependently attenuated by subsequent intra-NRM injection of the mu-opioid antagonist beta-funaltrexamine (beta-FNA), but not by the kappa-opioid antagonist nor-binaltorphimine (nor-BNI) or the delta-opioid antagonist naltrindole. The results demonstrated that oxytocin plays an antinociceptive role in the NRM of rats through activating the oxytocin receptor. Moreover, mu-opioid receptors, not kappa and delta receptors, are involved in the oxytocin-induced antinociception in the NRM of rats. PMID- 14556958 TI - Allatostatin gene expression in brain and midgut, and activity of synthetic allatostatins on feeding-related processes in the cockroach Blattella germanica. AB - Allatostatins of the YXFGLamide group were discovered in cockroaches through their capacity to inhibit juvenile hormone biosynthesis. Here, we assess the occurrence of preproallatostatin (preproAST) mRNA in the brain and midgut of adult females of the cockroach Blattella germanica, and estimate brain and midgut preproAST mRNA levels during the first reproductive cycle. Reverse transcription polymerase chain reaction (RT-PCR) shows that brain preproAST mRNA levels increase slightly during the gonadotrophic cycle, and remain high during ootheca transport. In the midgut, preproAST mRNA levels decline around the middle of the gonadotrophic cycle. The pattern of allatostatin expression in gut tissues suggests that these peptides play roles related to feeding and nutrition. Our results have shown that synthetic allatostatins inhibit hindgut motility and activate midgut alpha-amylase secretion. In addition, injected allatostatins inhibit food consumption, which might be connected to the above activities. PMID- 14556959 TI - Expression and characterization of the preoptic regulatory factor-1 and -2 peptides. AB - The preoptic regulatory factors, PORF-1 and PORF-2, were originally detected as cDNAs from the rat preoptic area of the hypothalamus. Expression of the messenger RNAs is dependent on age, gender and hormonal status. PORF-1 is a putative transcription factor while PORF-2 affects cell growth. However, expression of peptides predicted from the PORF-1 and PORF-2 open reading frames has not been reported. The characterization of PORF-1 and PORF-2 peptides expressed in vitro and in vivo is described here. There are three potential PORF-1 peptides of 37, 59 and 86 amino acids, because of the presence of two TGA codons that can code either for termination or for selenocysteine incorporation in the PORF-1 open reading frame. All three peptides are detected after in vitro translation of full length porf-1 brain mRNA. The two larger ones are found in normal male rat brain. The expression of a given peptide in bacteria depends upon the host strain. In contrast, a single 75 amino acid PORF-2 peptide is detected by in vitro translation, in transformed bacteria, and in rat brain. The PORF-1 and PORF-2 peptides have the charges and hydrophobicities predicted from their brain cDNA open reading frames. These small peptides may represent examples of multiple usage of single genetic loci in eukaryotes, giving rise to both a protein product and its regulatory peptide. PMID- 14556960 TI - Involvement of transmitters in pituitary adenylate cyclase-activating polypeptide induced hyperthermia. AB - The involvement of transmitters in the hyperthermic effect of centrally administered pituitary adenylate cyclase-activating polypeptide (PACAP-38) was studied. Rats were treated with different receptor antagonists or agonists in doses that per se proved to be ineffective. The following agonists and antagonists were used: haloperidol, phenoxybenzamine, propranolol, atropine, bicuculline, naloxone, apomorphine, bromocriptine and methysergide. Apomorphine and bromocriptine enhanced the elevation of body temperature induced by PACAP-38. The PACAP-38-hyperthermia was antagonized by haloperidol while other receptor blockers used were ineffective. Our results suggest that dopaminergic but not cholinergic, noradrenergic, serotoninergic, GABA-ergic or opioid mediation may be involved in the hyperthermic effect of PACAP-38 in rats. PMID- 14556961 TI - Bradykinin B1 receptor stimulates the proximal tubule Na(+)-ATPase activity through protein kinase C pathway. AB - Recently, our group described a B1-mediated stimulatory effect of des-Arg(9) bradykinin (DABK) on the Na(+)-ATPase activity of proximal tubule basolateral membranes (BLM) [Biochim. Biophys. Acta 1431 (1999) 483.]. Data in the present report suggest the participation of a phosphatidylinositol-specific PLC (PI PLC)/protein kinase C (PKC) pathway as the molecular mechanism of DABK-mediated stimulation of the Na(+)-ATPase activity since (i) 10(-8) M DABK activates PI-PLC activity; (ii) 10(-9) M U73122, a PI-PLC inhibitor, abolishes the effect of 10( 8) M DABK on the Na(+)-ATPase activity; (iii) 10(-8) M DABK increases phosphoprotein formation by 34%. This effect is completely reversed by 10(-7) M calphostin C, an inhibitor of PKC; (iv) 20 ng/ml TPA, an activator of PKC, and 10(-8) M DABK stimulate the Na(+)-ATPase activity in a similar and nonadditive manner. Furthermore, the effect of 10(-8) M DABK is completely reversed by calphostin C; (v) 10(-8) M DABK increases phosphoserine residue levels by 54%. This effect is completely reversed by 10(-7) M calphostin C. PMID- 14556962 TI - Sarcosine1,glycine8 angiotensin II is an AT1 angiotensin II receptor subtype selective antagonist. AB - Studies predating the discovery of the two major subtypes of angiotensin II (Ang II) receptors, AT1 and AT2, revealed anomalous characteristics of sarcosine1,glycine8 Ang II (Sar1,Gly8 Ang II). It competed poorly for 125I-Ang II binding in bovine brain but potently antagonized dipsogenic responses to intracerebroventricularly administered Ang II. Subsequent recognition that bovine brain contains AT(2) receptors, while dipsogenic responses to Ang II are mediated by AT1 receptors, suggests that Sar1,Gly(8) Ang II is AT1 selective. Sar1,Gly8 Ang II competed for 125I-sarcosine1,isoleucine8 Ang II binding to AT1 receptors in pituitary, liver and adrenal (the latter with the AT2 selective antagonist PD 123,319) with Ki's of 0.66, 1.40 and 1.36 nM, respectively. In contrast, the Ki of Sar1,Gly8 Ang II for AT2 receptors in rat adrenal (with the selective AT1 antagonist losartan) was 52 nM. 125I-Sar1,Gly8 Ang II (0.5-3 nM) bound to AT1 receptors in pituitary, liver, heart, adrenal, and hypothalamic membranes with high affinity (Kd=0.43, 1.6, 2.3, 0.96 and 1.8 nM, respectively), but showed no saturable binding to the adrenal AT2 receptor. 125I-Sar1,Gly8 Ang II selectively labeled AT1 receptors in sections of adrenal using receptor autoradiography. Thus, binding studies reveal Sar1,Gly8 Ang II to be the first angiotensin peptide analog to show AT1 receptor selectivity. 125I-Sar1,Gly8 Ang II offers a new means to selectively radiolabel AT1 receptors and may help to characterize ligand docking sites and agonist switches for AT1 versus AT2 receptors. PMID- 14556963 TI - Protective effect of bradykinin antagonist Hoe-140 during in vivo myocardial ischemic-reperfusion injury in the cat. AB - The effect of icatibant (Hoe-140), a selective bradykinin receptor (B(2)) antagonist on myocardial ischemic-reperfusion injury was studied in open chest barbiturate anaesthetized cats. The left anterior descending coronary artery was occluded for 15 min, followed by 60 min of reperfusion. Saline or icatibant (200 microg/kg) was administered intravenously slowly over 2 min, 5 min before reperfusion. In the saline-treated group, myocardial ischemic-reperfusion injury was evidenced by depressed MAP, depressed peak positive and negative dP/dt and elevated left ventricular end-diastolic pressure and enhanced oxidative stress [elevated plasma thiobarbituric acid reactive substances (TBARS; a marker for lipid peroxidation), depressed myocardial GSH (reduced glutathione), superoxide dismutase (SOD), catalase] and depletion of adenosine triphosphate (ATP) along with rise in plasma creatine phosphokinase (CPK). Administration of icatibant resulted in complete hemodynamic recovery together with repletion of ATP and reduction in plasma TBARS without any significant change in myocardial SOD, catalase and GSH. The results of the present study suggest a protective role of icatibant in myocardial ischemic-reperfusion injury. PMID- 14556964 TI - Low T cell receptor excision circle levels in patients thymectomized 25-54 years ago. PMID- 14556965 TI - Genetic and structural polymorphism of complement receptor 1 in normal Indian subjects. AB - The human complement receptor 1 (C3b/C4b receptor, CD35, CR1), a polymorphic membrane bound glycoprotein, is differentially expressed on erythrocytes, eosinophils, monocytes, B and T-lymphocytes, dendritic cells and kidney podocytes. It also occurs in the plasma as soluble CR1 (sCR1) and in urine as urinary CR1 (uCR1). Different population studies have suggested the functional and physiological significance of the structural (CR1-A/190, CR1-B/220, CR1-C/160 and CR1-D/250 kDa) and genomic (HH, high erythrocyte CR1 expression, HL, intermediate and LL, low expression) polymorphisms in health and disease. However, simultaneous study on the structural and genomic polymorphism in the same group of study subjects is lacking. This is the first study on both quantitative and structural polymorphism in 101 healthy volunteers from different parts of India by random sampling. In our study, AA phenotype was found to be expressed in 84.2% of individuals and 14.8% carried AB phenotype. One individual (0.9%) was found to possess BB phenotype. Homozygous BB pattern was identified for the first time in Indian subjects. The relative gene frequencies for A and B allele were found to be 0.916 and 0.084, respectively. Pertaining to quantitative polymorphism, percentage distribution for HH, HL and LL phenotypes was found to be 23.7, 54.45 and 21.79%, respectively and the gene frequencies were 0.51 and 0.49 for H and L allele, respectively. The observations for quantitative as well as structural polymorphism showed a good probability of fitness with Hardy Weinberg equilibrium, thus proving that both the types of CR1 polymorphic forms are encoded by autosomal co-dominant alleles. We found a higher frequency of HL and AA phenotypes in the study subjects. Our findings are unique as we found that gene frequencies for structural and quantitative polymorphism in our study subjects were a combination of those found in Caucasian and Oriental populations. PMID- 14556966 TI - Interleukin-12 release from macrophages by hyaluronan, chondroitin sulfate A and chondroitin sulfate C oligosaccharides. AB - Mixtures of hyaluronan (HA), chondroitin sulfate (CS)-A and CS-C oligosaccharides were generated through the enzymatic digestion of the polysaccharides with either mammalian hyaluronidase or bacterial HA lyase or chondroitinase. Compared to mammalian enzymes, bacterial enzymes hydrolyze the polysaccharides through a different mechanism yielding chemically distinct sets of oligosaccharides. Peripheral leukocytes and a human monocytic cell line were exposed to these oligosaccharides and the amount of interleukin-12 released by the cells was measured. For all types of oligosaccharide tested, we found that the amount of interleukin-12 induced by oligosaccharides generated with bacterial enzyme was significantly lower than the amount of interleukin-12 induced by oligosaccharides generated with mammalian enzyme. In addition, we observed that CS oligosaccharides generated with bacterial enzyme were capable of reducing the lipopolysaccharide-induced interleukin-12 production in macrophages. Our results indicate that HA or CS oligosaccharides generated with mammalian enzymes might possess pro-inflammatory potential, while HA or CS oligosaccharides generated with bacterial enzymes might possess non- or anti-inflammatory properties. The implications of our findings in view of the ongoing investigation of the potential therapeutic benefits of HA and CS in arthritis or other inflammatory pathologies are discussed. PMID- 14556967 TI - Induction of cell cycle regulatory proteins by murine B cell proliferating pectic polysaccharide from the roots of Bupleurum falcatum L. AB - Bupleuran 2IIc, a pectic polysaccharide isolated from the roots of Bupleurum falcatum L., was characterized as a T-cell-independent B cell mitogen, that activates, proliferates and differentiates B cells in vivo and in vitro (Immunology 97 (1999) 540). Studies were focused on elucidating the mechanism by which bupleuran 2IIc causes proliferation of B cells and expression of cell cycle regulatory proteins. B cells showed slower rates of entry into the S and G2/M phases of the cell cycle when stimulated with bupleuran 2IIc versus anti-IgM. However, the Stimulation Index continued up to two times longer with bupleuran 2IIc over anti-IgM. Although both bupleuran 2IIc and anti-IgM induced similar expressions of cell cycle regulatory proteins, cyclins D2, A, and B1, in B cells, those cells stimulated with bupleuran 2IIc appeared to sustain expressions of these protein for longer periods of time. Stimulation of B cells with bupleuran 2IIc induced phosphorylation of retinoblastoma protein, pRB, an important gene product regulating the restriction point, R, which is responsible for the transition from the G0/G1 to the S phases of the cell cycle. The results of this study demonstrate that both bupleuran 2IIc and anti-IgM interact with B cells, thus, leading to expressions of cell cycle regulatory proteins. However, the respective modes of binding and proximity of interactions with the B cell membrane may differ. PMID- 14556968 TI - The beta-globin HS4 insulator confers copy-number dependent expression of IgH regulatory elements in stable B cell transfectants. AB - Locus control regions (LCR) were first defined by their theoretical ability to enhance the expression of linked genes in a tissue-specific, position-independent and copy-number dependent manner. In fact, few of the so-called LCR identified completely fulfil this definition. For example, the regulatory elements located in 5' (Emu) and 3' (HS3a; HS1,2; HS3b; HS4) of the IgH locus display some properties of a LCR but lack a copy-number dependence and sometimes display position effects in transgenes. In order to study whether addition of insulators would allow to overcome such problems in transgenes, we studied constructs harboring a V(H) promoter-green fluorescent protein reporter gene linked to the 3' and/or 5' IgH elements, surrounded or not with the chicken beta-globin 5'HS4 insulator. When flanked with insulators it appeared that either 3' IgH and 5' IgH regulatory elements now behave as true LCR elements and noticeably display copy number dependence in transfected pre-B or B cell lines. PMID- 14556969 TI - MHC class I-mediated exogenous antigen presentation by exosomes secreted from immature and mature bone marrow derived dendritic cells. AB - Exosomes are 50-90 nm vesicles with antigen presenting ability carrying major histocompatibility complex (MHC) class I, class II, abundant co-stimulatory molecules and some tetraspan proteins. Although dendritic cells (DCs) are one of the professional antigen presenting cells capable of presenting exogenous antigens in MHC class I-mediated antigen specific manner (cross-presentation), the cross-presentation ability by exosomes from immature or mature DCs are unknown. Here we show that exosomes released from ovalbumin (OVA) protein-pulsed bone marrow derived dendritic cells (BM-DCs) weakly present the peptide determinants to OVA specific MHC class I-restricted CD8(+) T cell hybridomas. The exosomes secreted by OVA(257-264) peptide- or OVA protein-pulsed mature BM-DCs activated OVA specific MHC class I-restricted T cell hybridomas more efficiently than those from immature BM-DCs. Transporters associated with antigen processing (TAP) deficient mice-derived BM-DCs were also used to examine whether functional TAP activity was required for cross-presentation by exosomes. The exosomes obtained from OVA(257-264) peptide-pulsed BM-DCs derived from TAP(-/-) mice showed a significant antigen presenting ability to OVA specific MHC class I restricted T cell hybridomas. Altogether, our data indicate that BM-DCs secrete exosomes with weak cross-presentation ability. PMID- 14556970 TI - Stromal cell-derived factor-1 production by spleen cells is affected by nitric oxide in protective immunity against blood-stage Plasmodium chabaudi CR in C57BL/6j mice. AB - Malaria, a major endemic tropical disease, is caused by the infection of blood cells by Plasmodium protozoa. Most patients control their parasitemia by a not fully understood spleen-dependent mechanism. SDF-1alpha is a chemokine produced by stromal cells such as reticular spleen cells. Nitric oxide (NO) has several immune functions, including killing of intracellular pathogens and its function in malaria is debated. We have previously shown that SDF-1alpha production peaks during the ascending parasitemia in Plasmodium chabaudi infection and its supplementation in lethal models could reduce the parasitemia. In the present study, we analyzed SDF-1 production by spleen cells as related to NO metabolism in the P. chabaudi rodent malaria model using IFN-gamma; TNFR and iNOS-knockout mice or iNOS-blocked, L-NAME- or aminoguanidine-treated mice. Parasitemia and production of SDF-1alpha and SDF-1beta were determined by RT-PCR. In vitro NO production by spleen adherent cells was also tested. The data showed that parasitemia was less intense in both iNOS(-/-) or NO-inhibited mice than in controls, with increased and long-lasting production of SDF-1alpha mRNA. In the absence of cytokines involved in the final regulation of NO production by effector cells, as is the case for TNFR(-/-) and GKO mice, the infection progressed in an uncontrolled manner regardless of SDF-1alpha production, suggesting that these cytokines must be involved in the control of parasitemia after the SDF-1alpha dependent process. The SDF-1beta isoform was constitutive in all experiments, with elevated levels only clearly seen in TNFR(-/-) mice. We conclude that SDF-1 is involved in the promotion of parasitemia control in malaria, and excessive NO could affect its production. PMID- 14556971 TI - Macrophage distinguishes Vibrio cholerae hemolysin from its protease insensitive oligomer by time dependent and selective expression of CD80-CD86. AB - The monomeric and oligomeric forms of Vibrio cholerae hemolysin (HlyA), a membrane damaging toxin that forms transmembrane pentameric diffusion channels in target eukaryotic membrane, show a pronounced difference in protease susceptibility, presumably due to masking of sensitive peptide bonds during oligomerization. In this work, we examined if resistance of a protein to proteolytic processing affects the expression of costimulatory molecules, CD80 and CD86, on macrophage exposed to the same antigen. The murine peritoneal cavity macrophages expressed both CD80 and CD86 after 24 h of incubation with HlyA monomer but failed to express the costimulatory molecules when treated with the HlyA oligomer. The expression of CD80 molecule on macrophage after 48 h by the HlyA oligomer that failed to express the costimulatory molecules after 24 h indicates that proteolytic processing plays a decisive role in expression of CD80 and CD86 on cell surface. PMID- 14556972 TI - Differential role of mitogen-activated protein kinases in CD40-mediated IL-12 production by immature and mature dendritic cells. AB - Using a murine spleen-derived dendritic cell (DC) line (BC1) CD40-mediated interleukin (IL)-12 production was analyzed and compared between immature and mature DC. BC1 cells, immature DC (iDC), were maturated by treatment with lipopolysaccharide (LPS) or tumor necrosis factor (TNF)-alpha. IL-12 production of LPS-treated DC (LPS/DC) was markedly enhanced by treatment with an anti-CD40 monoclonal antibody (mAb). Although the anti-CD40 mAb also enhanced IL-12 productions of iDC and TNF-alpha-treated DC (TNF/DC), these production levels were considerably low compared with that of LPS/DC. CD40-mediated IL-12 productions by iDC and TNF/DC were significantly enhanced by treatment with PD98059, a specific inhibitor of extracellular signal-related kinase (ERK) pathway. In contrast, PD98059 showed no significant effects on CD40-mediated IL 12-production by LPS/DC. These results demonstrated that ERK pathway was involved in negative regulation of the IL-12 productions by iDC and TNF/DC but not by LPS/DC. On the other hand, SB203580, a specific inhibitor of p38 mitogen activated protein kinase (MAPK) pathway, completely inhibited CD40-mediated IL-12 production by iDC, while not affecting those of TNF/DC and LPS/DC. Thus, p38 MAPK pathway appeared to positively regulate the IL-12 production in iDC but not in mature DC. It seems that roles of ERK and p38 MAPK for IL-12 production are developmentally changed in murine DC. PMID- 14556973 TI - Approaches toward reversal of increased vascular permeability in C1 inhibitor deficient mice. AB - C1 inhibitor (C1INH) deficient mice have increased vascular permeability that can be demonstrated by the extravasation of Evans Blue dye. This vascular leak is reversed with protease inhibitors, such as C1INH itself, DX88 (a recombinant variant Kunitz domain plasma kallikrein inhibitor), and the bradykinin receptor type 2 antagonist, Hoe140. The studies described here were undertaken for the following reasons: (1) To provide a more quantitative analysis of the effects of these interventions; (2) to provide data to further test the hypothesis that increased vascular permeability results from contact system activation with kallikrein-mediated release of bradykinin; (3) to test the hypothesis that the amino terminal non-serpin domain of C1INH modulates access to complex proteases, such as kallikrein complexed with high molecular weight kininogen (HK); and (4) to determine whether attenuated androgens or estrogens exert a direct effect on C1INH synthesis. To characterize the differences in these reagents, the dose response and the rate of reappearance of increased vascular permeability in C1INH(-/-) mice were determined for the following agents: human plasma-derived C1INH, a recombinant Kunitz domain plasma kallikrein inhibitor (DX88), a bradykinin receptor antagonist (Hoe140), and a recombinant C1INH with an amino terminal truncation at amino acid 98 and substitution of the P2 Ala with a Val (Cserp98,A443V). C1INH and Cserp98,A443V were equivalent in activity, which provides further support for the hypothesis that the vascular leak is mediated by bradykinin and suggests that the amino terminal domain neither enhances nor interferes with access to kallikrein within the kallikrein-HK complex. DX88 was effective at very low doses, as was Hoe140. The duration of action of Hoe140 was quite prolonged. The data indicate that, in the mouse, neither danazol nor estrogens have a significant effect on C1INH synthesis. PMID- 14556974 TI - Human chymase stimulates Ca2+ signaling in human polymorphonuclear cells. AB - Human chymase is known to function as a chemoattractant for human leukocytes. To investigate the mechanism of the chymase-induced cell migration, change in intracellular calcium concentration ([Ca(2+)]i) was examined in human polymorphonuclear (PMN) cells using Fluo-3 as a fluorescent Ca(2+) indicator. Treatment of PMN cells with human chymase caused [Ca(2+)]i elevation in a concentration-dependent manner. Depletion of extracellular Ca(2+) from the medium partially attenuated the chymase-induced [Ca(2+)]i increase, showing that both Ca(2+) influx and Ca(2+) release from internal stores might be involved in the [Ca(2+)]i response. Pretreatment of the cells with pertussis toxin completely blocked the chymase-induced [Ca(2+)]i signal, suggesting an involvement of G protein in the chymase-mediated [Ca(2+)]i elevation. The data in the present study raise the possibility that the chymase-induced cell migration is mediated by the [Ca(2+)]i elevation, which might be caused by stimulation of a G-protein coupled receptor such as protease-activated receptors (PARs). PMID- 14556975 TI - Effective elicitation of anti-tumor immunity by collocation of antigen with encoding gene in the same vaccine. AB - Peptide and naked DNA vaccines, aimed at generating strong cytotoxic T lymphocyte (CTL) responses capable of mediating tumor regression, have been considered of the most rapidly evolving technologies for tumor vaccination. Results from clinical trials of these strategies are encouraging, but unfortunately, most of those trials have been proved as partly successful. Given the distinct dynamics of antigen presentation for peptide and gene forms of antigens, we explored a novel concept that collocation of the antigen with the encoding gene in the same vaccine could effectively elicit anti-tumor immunity, and developed a novel peptide-DNA dual vaccine (PDDV), which combines the benefits of peptide and DNA vaccines and could induce tumor-specific CTL response. Furthermore, PDDV effectively protected mice against fatal P815 tumor challenge and cured tumor bearing DBA/2 mice, suggesting PDDV as a potential formulation of tumor vaccine. PMID- 14556976 TI - Effect of an acidic heteropolysaccharide (ARAGAL) from the gum of Anadenanthera colubrina (Angico branco) on peritoneal macrophage functions. AB - Brazilian flora are a source of interesting polysaccharides which, either in their native state or when submitted to structural modifications, might have potential applications as biological response modifiers (BRM). A complex acidic heteropolysaccharide, containing mainly galactose and arabinose (ARAGAL), isolated from the gum of the native leguminous tree Anadenanthera colubrina (Angico branco), was studied for its immunological properties on peritoneal exudate cells, namely their superoxide anion production, phagocytic activity, morphological alterations and percentage content of activated macrophages. Activation of macrophages showing increased cytoplasm, bright and large nuclei, various cytoplasmatic projections and spreading ability, was detected following in vitro cell exposure to ARAGAL or in cells obtained from treated animals. In vitro exposure to ARAGAL increased the occurrence of activated macrophages in a time- and a dose-dependent pattern, since approximately 82% of the cells were activated in the presence of 300 microg/ml of ARAGAL after 24 h of incubation and approximately 91% after 48 h. The occurrence of activated macrophages was also evident in cell preparations from ARAGAL-treated mice, their percentage showing a dose-dependent pattern. There were approximately 60, 75 and 75% following treatment with 100, 250 and 500 mg/kg of ARAGAL, respectively. A phagocytic assay showed that 25 microg/ml ARAGAL was sufficient to impose a maximum phagocytic ability, although this effect was dose-dependent. O(2)(-) production by macrophages from ARAGAL-treated mice was 70% higher than that of cells from untreated mice. Moreover, cells from treated mice responded to PMA, the effect being 25% higher than that of the control using untreated mice. These results thus suggest a possible role of ARAGAL from A. colubrina as a BRM. PMID- 14556977 TI - Prostanoids and MPO-halide system products as a link between innate and adaptive immunity. AB - and The crosstalk between innate adaptive immunity is regulated by cytokines and complex interactions between cells of the immune system. A variety of endogenous agents are involved in the regulation of the cytokine network. Especially, eicosanoids and ROIs have a great impact on the regulation of cytokine production. Eicosanoids (prostanoids, leukotrienes and lipoxins) are produced mainly by inflammatory cells while their receptors are distributed on the cells of both arms of the immune system. Depending on the predominant prostanoid produced and the profile of prostanoid receptors expression on immune cells, eicosanoids can selectively regulate the production of Th1 and Th2 driven cytokines. Inflammatory cells (neutrophils, macrophages), are also a rich source of large amounts of ROIs. In this paper we have focused on the role of taurine chloramine (TauCl), the physiological product of neutrophil MPO-halide system, in the regulation of immune system. It is well documented that TauCl has pleiotropic effects on the inductive phase of the immune response. TauCl's immunoregulatory properties result from its ability to modulate the production of cytokines and eicosanoids. Finally, we conclude that eicosanoids and ROIs provide an important link between the afferent branches and the innate and adaptive immune response. PMID- 14556978 TI - Involvement of ICOS-B7RP-1 costimulatory pathway in the regulation of immune responses to Leishmania major and Nippostrongylus brasiliensis infections. AB - The ICOS-B7RP-1-mediated T cell costimulatory pathway has been implicated crucial for T cell activation and differentiation. In this study, we investigated the role of this costimulation in the regulation of immune responses to parasitic infections by using blocking antibody against B7RP-1 as well as ICOS-deficient mice. The administration of anti-B7RP-1 monoclonal antibody (mAb) significantly suppressed the footpad swelling in susceptible BALB/c mice upon Leishmania major infection. The observation was consistent not only with the significant suppression of IL-4, IL-5 and IL-10 secretion from lymph node cells, which were derived from L. major-infected mice, but also with the significant reduction of total serum IgE and IgG(1) in anti-B7RP-1 mAb-treated BALB/c mice. Infection of ICOS-deficient mice with L. major also suggested the impaired Th2 immune responses in the absence of this costimulation. The immunological function of ICOS-B7RP-1 costimulatory pathway in infection was further confirmed by infecting anti-B7RP-1 mAb-treated wild type or ICOS-deficient mice with Nippostrongylus brasiliensis. The characteristic elevation of total serum IgE and eosinophilia upon N. brasiliensis infection was suppressed by blocking this costimulation. Moreover, the protection to N. brasiliensis adult worms was suppressed in anti B7RP-1 mAb-treated wild type or ICOS-deficient mice. These results suggest the crucial role of this costimulatory pathway in the regulation of Th2-biased T cell differentiation and in host immune responses against L. major and N. brasiliensis infections. PMID- 14556979 TI - Toll-like receptor 4 is not involved in host defense against respiratory tract infection with Sendai virus. AB - Toll-like receptors (TLR) induce innate immune responses upon stimulation by a wide variety of pathogens. TLR4 has been implicated in innate immunity against respiratory syncytial virus (RSV) by an interaction with the viral envelope fusion (F) protein. Sendai virus (mouse parainfluenza type 1) shares many features with RSV, including a structurally and functionally similar F protein. To determine the role of TLR4 in host defense against Sendai virus respiratory tract infection, TLR4 mutant and wildtype mice were intranasally infected with Sendai virus. Sendai infection resulted in an increase in viral RNA copies in lung homogenates peaking on day 4. Pulmonary viral loads, histopathology, cytokine levels and leukocyte influx were similar in TLR4 mutant and wildtype mice. In spite of the structural similarities shared by the F proteins of Sendai virus and RSV, TLR4 is not involved in host defense against respiratory tract infection with Sendai virus. PMID- 14556980 TI - Attenuated nef DNA vaccine construct induces cellular immune response: role in HIV-1 multiprotein vaccine. AB - HIV-1 positive patients generate Nef-specific CTL response, indicating that Nef is a potent immunogen. However, Nef is also known to down regulate the expression of CD4 and MHC-I molecules, thereby protecting virally infected target cells. We compared the immunogenicity of non-functional nef vaccine constructs to wild type functional nef as potential immunogen. Mice were immunized with different nef constructs and assessed for their ability to induce cellular immune responses. Evaluation of T cell immune responses in mice showed that non-functional nef vaccine constructs are capable of inducing a significant T cell immune response measured by IFN-gamma ELISPOT. Further epitope mapping studies indicate that one of our attenuated constructs, Nef R-38, has multiple CTL epitopes spanning throughout the gene. Our results indicate that functionally attenuated Nef antigen might be a better candidate for future multiprotein HIV-1 vaccine. PMID- 14556981 TI - Modulation of the constitutive or cytokine-induced bronchial epithelial cell functions in vitro by fluticasone propionate. AB - When exposed to proinflammatory mediators, human bronchial epithelial cells (HBECs) upregulate the 'constitutive' adhesion molecule expression and cytokine/chemokine release. We tested whether and to what extent the inhibitory effect of fluticasone propionate on HBECs could involve the 'constitutive' and 'cytokine-induced' proinflammatory functions. Stimulation of the HBECs with interleukin (IL)-4 plus tumour necrosis factor (TNF)-alpha was more effective in upregulating intercellular adhesion molecule (ICAM)-1 ( approximately 2.2-fold increase) than vascular adhesion molecule (VCAM)-1 ( approximately 1.6-fold increase) expression (P<0.05) and in increasing the release of 'regulated on activation normal T cell expressed' (RANTES, 5.7-fold increase) than of IL-8 (3.5 fold increase) and granulocyte macrophage-colony stimulating factor (GM-CSF, 2.8 fold increase), (P<0.01). Fluticasone propionate, at the two concentrations tested (10 and 100 nM), was more effective in inhibiting the 'IL-4 plus TNF-alpha induced' ICAM-1 expression than VCAM-1 expression (P<0.05) and in downregulating RANTES than IL-8 or GM-CSF secretion (P<0.05). The degree of inhibition demonstrated by fluticasone propionate appeared to be related to the degree of cell activation. In addition, for both adhesion molecules, the effect of fluticasone propionate at both concentrations tested appeared to be related to a complete inhibition of 'IL-4 plus TNF-alpha-induced' expression with no involvement of the 'constitutive' expression. Slightly different results were observed for cytokine/chemokine release. Indeed, evaluating RANTES, a complete inhibition of the 'IL-4 plus TNF-alpha-induced' release with a partial inhibition also of the 'constitutive' release at both concentrations of the drug tested was found, whereas for GM-CSF and IL-8, only a partial inhibition of the 'IL-4 plus TNF-alpha-induced' release in the presence of fluticasone propionate 10 and 100 nM. Thus, HBECs can constitutively or upon activation express adhesion molecules and secrete proinflammatory proteins at various levels and the different ability of fluticasone propionate to modulate the HBEC functions appears to be mostly related to the different inhibition of the various 'IL-4 plus TNF-alpha-induced' responses. PMID- 14556982 TI - Parenchymal cells in immune and tolerance induction. AB - There is increasing evidence that dendritic cells are capable of inducing T cell tolerance to tissue-specific antigens by presenting these antigens to CD4 and CD8 T cells in the respective draining lymph nodes. In contrast, parenchymal cells are often seen as immunologically inert. This lecture summarizes studies showing that tissue cells like sinusoidal endothelial cells and hepatocytes in the liver as well as keratinocytes in the skin can induce peripheral T cell tolerance. This tolerance induction is often preceded by T cell activation. Thus, several tolerance mechanisms are operating in parallel. We still have no learn which mechanism is most suitable for therapeutic intervention in autoimmune diseases and organ transplantation. PMID- 14556983 TI - Chlamydophila (Chlamydia) pneumoniae induces histidine decarboxylase production in the mouse lung. AB - Chlamydophila (Chlamydia) pneumoniae (C. pneumoniae) is the third most common cause of community-acquired pneumonia and is probably involved in the development of certain chronic inflammatory diseases, including atherosclerosis and adult onset asthma. Histamine, synthesized by histidine decarboxylase (HDC) from L histidine, plays an essential role in allergic and inflammatory processes and in cell differentiation. The effect of C. pneumoniae infection on the expression of HDC has not been examined. In the present study, normal Balb/c mice and HDC knockouts, and control mice with a CD1 background were infected intranasally with C. pneumoniae. On days 1, 3, 7, 16 and 31 after infection, the normal Balb/c mice were sacrificed and divided into three groups. In the homogenized lungs of the first group, C. pneumoniae titres were determined and demonstrated peak levels on day 7. HDC production was revealed by a Western blot assay throughout the observation period of 1-16 days, and cytokine concentrations were determined by ELISA. The interleukin-3 (IL-3) and interleukin-6 (IL-6) levels were highest on day 1 and on days 1-3, respectively; the interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) levels reached the maximum on day 7, but the quantity of IL 4 was still three times higher than that in the control group 16 days after infection. The lungs of the mice in the second group were processed for the in situ demonstration of HDC activity, while the lungs in the third group were stained for C. pneumoniae antigen. The HDC activity was increased predominantly in the bronchial epithelial cells, while C. pneumoniae antigens were expressed especially in the interstitial macrophages. The HDC knockout mice exhibited a higher survival rate after C. pneumoniae infection than did the control mice. These results point to a strong association between local histamine production and other inflammatory mediators and are novel in demonstrating the role of histamine in the pathomechanism of C. pneumoniae infections. PMID- 14556984 TI - Current issues in antigen presentation--focus on the dendritic cell. AB - A personal view of some of the major current issues in antigen presentation, which focuses on the biology of the dendritic cell. The question of dendritic cell heterogeneity is discussed and the inclusion of the "plasmacytoid" dendritic cell in this family is questioned. Defining the molecules involved in dendrite formation may ultimately provide the only reliable signature of the dendritic cell. The role of the dendritic cell as a key link between innate and adaptive immunity is here revisited, and the direct and indirect interactions between dendritic cells and microorganisms are discussed. Reactive oxygen species may be key modulators of dendritic cell function, although it remains unclear whether direct or indirect mechanisms are more important. Study of the cell biology of antigen processing reveals a tightly regulated complex system involving multiple proteinases, adapted to the most efficient presentation of invading microorganisms and playing a key role in shaping the T cell repertoire. PMID- 14556985 TI - Immunity to blood-stage murine malarial parasites is MHC class II dependent. AB - To determine whether MHC class II antigen presentation is essential for the induction of protective immunity against blood-stage malarial parasites, we used gene-targeted knockout (KO) mice to follow the time-course of nonlethal Plasmodium yoelii and Plasmodium chabaudi infections in two models of MHC class II deficiency. Infection of MHC class II KO (A(-/-)) mice with either parasite species resulted in an unremitting hyperparasitemia, whereas MHC-intact control mice resolved their parasitemia. In contrast, invariant chain KO (Ii(-/-)) mice, which present antigen via recycled but not nascent MHC class II molecules, eventually cured their infections when infected with P. yoelii. P. chabaudi parasitemia declined to subpatent levels in most Ii(-/-) mice but then recrudesced. Immunity to blood-stage malaria may be achieved by cell-mediated and antibody-mediated mechanisms of immunity, as such, the findings in A(-/-) mice indicate an essential role for MHC class II presentation of malarial antigens. Moreover, they suggest that protective immune responses to malarial antigens capable of eliminating blood-stage parasites are T cell dependent and can be induced with antigens processed in early and late endosomes. PMID- 14556986 TI - CD27 synergizes with CD40 to induce IgM, IgG, and IgA antibody responses of peripheral blood B cells in the presence of IL-2 and IL-10. AB - CD40, a member of the tumor necrosis factor receptor (TNFR) family, promotes IgM, IgG, and IgA antibody (Ab) synthesis in combination with a variety of cytokines. Another TNFR family member, CD27, causes B cells to differentiate into antibody forming cells, with marginal effects on proliferation. In the present study, we examined whether anti-CD27 monoclonal antibody (mAb) modulates the antibody production induced by anti-CD40 mAb immobilized on L cells expressing FcgammaRII (CDw32) in the presence of IL-2 and/or IL-10. The anti-CD40 mAb substantially enhanced IgM, IgG, and IgA production in combination with IL-2 and IL-10, whereas anti-CD27 mAb augmented it only marginally, as assessed by enzyme-linked immunosorbent assay. The addition of anti-CD27 mAb enhanced the anti-CD40 mediated IgM, IgG, and IgA antibody production only when both IL-2 and IL-10 were present in the culture. The CD27-positive B cell compartment generated synergistic antibody responses in response to four different stimulants, anti CD27/anti-CD40 mAb and cytokines IL-2/IL-10, whereas the CD27-negative B cell compartment failed to do so. Kinetic analysis showed that anti-CD40 might function in the early phase of B cell activation, while anti-CD27 mAb functioned in the late stage. The addition of CD27(-) to CD27(+) B cells in various ratios did not have any effect on the antibody production, suggesting that CD27(+) to CD27(-) B cell interaction does not occur in this system. Our findings suggest that a member of the TNFR family, CD27, cooperates with CD40 to induce efficient antibody production in combination with cytokines IL-2 and IL-10. PMID- 14556987 TI - Immunotolerance toward native alphaA-crystallin in knockout mice deficient in the functional protein. AB - Immune response against self antigens is normally prevented by an elaborate immunotolerance mechanism. A potential problem for recipients of gene therapy is, therefore, an immune response against the newly introduced gene product. To examine this issue we tested the immune response to the native proteins in knockout (KO) mice in which the genes for alphaA- or alphaB-crystallin were disrupted by partial or complete gene deletion, respectively. alphaA- and alphaB crystallins are two immunologically distinct polypeptides which form the large ( approximately 800 kDa) complex in the lens referred to as alpha-crystallin. When immunized with murine alpha-crystallin, alphaB-crystallin KO mice, in which the corresponding gene was completely deleted, responded well to the absent self antigen. In contrast, alphaA-crystallin KO mice, with the partial gene deletion, resembled wild type (WT) mice in being immunotolerant toward the native crystallin. Although no functional alphaA-crystallin could be detected in the lens of alphaA-crystallin KO mice, mRNA transcript coding for a truncated alphaA crystallin gene was found in thymi of these mice, suggesting that thymic expression of a residual fragment of the protein is responsible for the tolerance induction. These data suggest that nonfunctional proteins may induce immunotolerance and protect recipients of gene therapy from immunity against the native proteins. PMID- 14556988 TI - Differential expression of stem cell mobilization-associated molecules on multi lineage cells from adipose tissue and bone marrow. AB - Our laboratory has characterized a population of stromal cells obtained from adipose tissue termed processed lipoaspirate cells (PLAs). PLAs, like bone-marrow derived mesenchymal stem cells (BM-MSCs), have the capacity to differentiate along the adipogenic, osteogenic, chondrogenic, and myogenic lineages, In order to better characterize these two multi-lineage populations, we examined the surface phenotype of both bone marrow and adipose tissue-derived cells from five patients undergoing surgery. PLA and BM-MSC cells were isolated, subcultivated, and evaluated for cell surface marker expression using flow cytometry. PLA and BM MSC cells both expressed CD13, CD29, CD44, CD90, CD105, SH-3, and STRO-1. Differences in expression were noted for cell adhesion molecules CD49d (Integrin alpha4), CD54 (ICAM-1), CD34, and CD106 (VCAM-1). While markedly similar, the surface phenotypes of PLA and BM-MSC cells are distinct for several cell adhesion molecules implicated in hematopoietic stem cell homing, mobilization, and proliferation. PMID- 14556989 TI - Candida albicans clades. AB - DNA fingerprinting with the complex probe Ca3 has revealed the following five Candida albicans clades: group I, group II, group III, group SA and group E. These groups exhibit geographical specificity. Group SA is relatively specific (i.e., highly enriched) to South Africa, group E is relatively specific to Europe, and group II is absent in the Southwest USA and South America. The maintenance of deep-rooted clades side by side in the same geographical locale and the apparent absence of subclade structure suggest little recombination between clades, but higher rates of recombination within clades. Exclusive 5 fluorocytosine resistance in the majority of group I isolates reinforces the above conclusions on recombination, and demonstrates that clades differ phenotypically. The ramifications of these findings with regard to pathogenesis are discussed. In particular, these findings lay to rest the idea that one strain represents all strains of C. albicans, support the need for a worldwide analysis of population structure and clade-specific phenotypic characteristics, and demonstrate that in the future, pathogenic characteristics must be analyzed in representatives from all five clades. PMID- 14556990 TI - Advances in antibody-mediated immunity against Mycobacterium tuberculosis: implications for a novel vaccine strategy. AB - Cell-mediated immunity is considered to be the major component of the host response against Mycobacterium tuberculosis, whereas antibody-mediated immunity historically has been considered inconsequential. In recent years, studies from several groups have challenged the traditional dogma and demonstrated that monoclonal antibodies can modify various aspects of mycobacterial infections. This review describes the experimental evidence supporting a role for antibodies in defense against mycobacterial infections and outlines future challenges to the field of antibody-mediated immunity against M. tuberculosis, with particular emphasis on the implications of these findings for a novel vaccine strategy. PMID- 14556991 TI - The phototoxicity of phenothiazinium derivatives against Escherichia coli and Staphylococcus aureus. AB - Phenothiazinium dyes, and derivatives, were tested for toxicity to Escherichia coli and Staphylococcus aureus. The dyes were generally lipophilic (log P>1) and showed inherent dark toxicity (minimum lethal concentrations: 3.1-1000 microM). Dye illumination (total light dose of 3.15 J cm(-1) over 30 min) led to up to eight-fold reductions in minimum lethal concentrations. Most of the illuminated dyes showed significant relative singlet oxygen yields (phi'delta: 0.18-1.35) suggesting a type II mechanism of generating a phototoxic response. Although generally up to six-fold more effective against S. aureus, the dyes tested efficiently killed E. coli and may be of particular use in combating Gram negative pathogens. PMID- 14556992 TI - Carriage of Neisseria meningitidis and Neisseria lactamica in northern Greece. AB - In response to an increase in the number of cases of invasive meningococcal disease (IMD) in northern regions of Greece, a survey was carried out to determine if there was an increase in carriage of Neisseria meningitidis, particularly in areas where there have been increases in immigrant populations from neighbouring countries. The second objective was to determine if there was an increase in the serogroup C:2a:P1.5,2 a phenotype associated with recent outbreaks or changes in antibiotic sensitivities. As carriage of Neisseria lactamica is associated with development of natural immunity to IMD, the third objective was to determine the carriage rate of N. lactamica in this population. Among 3167 individuals tested, meningococci were isolated from 334 (10.5%). Compared with our previous studies, the proportion of meningococcal carriers was significantly increased among children in secondary education (11.3%) (chi2=9.67, P<0.005) and military recruits (37.4%) (chi2=21.11, P<0.000). Only 5/334 (1.5%) isolates expressed the phenotype associated with the increase in IMD in Greece. N. lactamica was isolated from 146/3167 (4.6%) participants. It was isolated from 71/987 (7.2%) children attending primary or nursery schools; however, the highest proportion of carriers (11.3%) was found in the boarding school for young Albanian men. In the 21-59-year age range, the majority of N. lactamica isolates (22/25, 88%) were from women, probably due to closer or more prolonged contact with children in the primary school age range. Smoking was significantly associated with isolation of meningococci from men but not from women. Penicillin insensitive strains (25/334, 7.5%) were identified in all four regions examined; the majority (14/25, 56%) were obtained from military personnel. We conclude that there was a higher proportion of carriers in the population of northern Greece; however, the increase in carriage rate was not associated with the influx of immigrants from neighbouring countries, and there was not a higher incidence of the C:2a:P1.5,2 strain responsible for increased disease activity in Greece in either the immigrant or local populations. PMID- 14556993 TI - Evaluation of non-culture diagnosis of invasive meningococcal disease by polymerase chain reaction (PCR). AB - Antibiotic treatment prior to transport or admission to hospital has reduced the proportion of cases of invasive meningococcal disease (IMD) from which Neisseria meningitidis can be isolated by standard microbiological techniques. Identification of meningococci by polymerase chain reaction (PCR) was assessed in relation to microbiological diagnosis for cases over a 4-year period between 1998 and 2001. A screening assay for the IS1106 gene was used to detect meningococcal DNA and five additional assays for siaD and orf-2 genes were performed to determine the serogroup. PCR results were compared with results of bacteriological culture, other laboratory test results and clinical data. The sensitivity of the PCR assay for culture-confirmed cases was 98.5%. The specificity of the assay was 96% based on test results for patients from whom other bacteria were isolated, children with viral meningitis and afebrile negative controls. The siaD B/C/W-135 and Y as well as the orf-2 gene for serogroup A PCR assays were able to determine the serogroup for 75.2% of cases that were positive by PCR screening assay. When isolates from patients with IMD were tested by both agglutination and PCR, the results agreed in all cases. PCR is a useful tool for diagnosis of IMD when Gram stain and culture tests are negative due to antibiotic treatment prior to collection of samples for microbiological analyses. PMID- 14556994 TI - Pseudomonas aeruginosa-specific IgG1 and IgG2 subclasses in enhancement of pulmonary clearance following passive immunisation in the rat. AB - Pseudomonas aeruginosa is an opportunistic Gram-negative pathogen, which causes serious debilitating infections in patients with compromised lung function. The mechanism by which P. aeruginosa is cleared from the lung is not fully defined, although our previous studies have established a role for cellular immunity in protection against P. aeruginosa infections. This study aimed to evaluate the role of P. aeruginosa-specific IgG in protection against P. aeruginosa in a rat model of acute pulmonary infection. Immunoaffinity chromatography was used to purify total rat IgG from rat immune serum (rats immunised with P. aeruginosa) and non-immune serum. Untreated recipient rats were injected intravenously with different concentrations of pure IgG prepared from serum of unimmunised rats (non immune IgG) or from rats immunised intestinally with killed P. aeruginosa (immune IgG) and infected intratracheally with P. aeruginosa 18 h later. The protective capability of the purified IgG against P. aeruginosa was assessed by measurement of reduction in P. aeruginosa infection in the lung 4 h after instillation of bacteria. Enhanced bacterial clearance induced by IgG was determined to be dose dependent with a 1 mg dose failing to enhance clearance, whereas 5 mg of immune IgG enhanced clearance from the airways and the lung tissue. Measurement of the IgG1, IgG2a and IgG2b isotypes in serum and the lung lavage following transfer of P. aeruginosa-specific IgG found that all three were present. These results demonstrate that anti-P. aeruginosa IgG can enhance bacterial clearance from the airways in an acute infection and identify an important role for IgG in acute respiratory infections caused by P. aeruginosa. PMID- 14556995 TI - Escherichia coli bind to urinary bladder epithelium through nonspecific sialic acid mediated adherence. AB - The first step in the bacterial colonization and infection of uropathogenic Escherichia coli is adherence to uroepithelium. Over 80% of all urinary tract infections are caused by E. coli. Uropathogenic E. coli express several adherence factors including type 1 and P fimbriae, which mediate attachment to the uroepithelium through specific binding to different glycoconjugate receptors. We showed that P and type 1 fimbriae are not the sole adhesins on uropathogenic E. coli and sialic acid also mediates nonspecific bacterial adherence of uropathogenic E. coli and urinary bladder epithelium. PMID- 14556996 TI - Modulation of the humoral immune response of swine and mice mediated by toxigenic Pasteurella multocida. AB - Progressive atrophic rhinitis is an upper respiratory tract disease of pigs caused by toxigenic strains of the bacterium Pasteurella multocida. In this study the effect of P. multocida on the humoral immune response of pigs and mice was investigated. Pigs were given live intranasal challenge with either a toxigenic strain or a non-toxigenic strain of P. multocida, or were given daily intranasal instillation of a cell-free lysate of the toxigenic strain. Mice were given a live intranasal challenge of either a toxigenic or a non-toxigenic strain of P. multocida. All of the animals were immunised with ovalbumin and serum concentrations of anti-ovalbumin antibodies were quantified and compared between different treatment groups and control animals. Intranasal challenge with toxigenic P. multocida caused a significant reduction in the levels of anti ovalbumin IgG in both species. A similar effect was seen in pigs given a cell free extract of toxigenic P. multocida. Whilst the mechanism of this suppression is unclear, we surmise that immunomodulation of the host is an important virulence factor for toxigenic P. multocida, and could be an important function of the toxin. This immunomodulatory effect may enhance colonisation of P. multocida aiding horizontal transmission and may predispose to concurrent infection with other potential pathogens. PMID- 14556997 TI - Kinetic study of porcine GM-CSF expression in porcine alveolar macrophages and spleen cells. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) is an important regulator in inducing differentiation and proliferation of immune cells. The functional roles of porcine GM-CSF (pGM-CSF) have not yet been revealed. Therefore, expression patterns of pGM-CSF were investigated in immune cells after cloning and sequencing of whole pGM-CSF cDNA. Whole cDNA of pGM-CSF was amplified from porcine alveolar macrophages stimulated by lipopolysaccharide (LPS), using 5'- and 3'-rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR) methods. The products of 5'- and 3'-RACE-PCR were cloned, and the nucleotide sequence of whole pGM-CSF cDNA was determined (GenBank accession number AY116504). The kinetics of pGM-CSF mRNA expression were studied in porcine immune cells such as alveolar macrophages and spleen cells, using a real-time quantitative PCR. The expression of pGM-CSF in LPS-, phytohemagglutinin (PHA)-, or concanavalin A (ConA)-stimulated cells was always higher as compared to the control cells. The expression levels of pGM-CSF in alveolar macrophages were highest at 5 h after LPS stimulation and then continuously decreased in the late phase. In spleen cells, the LPS-stimulated group showed the highest levels after 5 h, but the PHA- and the ConA-stimulated groups showed slightly increased expression levels at the early phase and peaked at 24 h. To our knowledge, this is the first published report describing the nucleotide sequence of whole cDNA and the expression pattern of pGM-CSF using real-time quantitative PCR. These results indicate that pGM-CSF has its own characteristic expression profile in different immune cells. PMID- 14556998 TI - The intrahepatic immune response during chronic hepatitis B infection can be monitored by the fine-needle aspiration biopsy technique. AB - Frequent analysis of the intrahepatic cellular immune response during chronic hepatitis B infection is not feasible with the liver tissue biopsy technique, due to its risk profile and patient discomfort. We investigated whether the relatively safe and patient-friendly cytological fine-needle aspiration biopsy (FNAB) technique is suited for this purpose. FNABs taken during hepatitis flares in three chronic hepatitis B patients treated with interferon-alpha, showed significant increments of CD8(+)-lymphocytes compared with the FNABs taken before and after the flares. No increments were observed in peripheral blood. The increments of intrahepatic CD8+ lymphocytes detected by the FNAB were related to anti-viral immune reactivity, since they coincided with significant serum hepatitis B virus DNA level reductions and in two of three patients with HBeAg seroconversion. In conclusion, the FNAB technique is suited to investigate the intrahepatic immune response during chronic hepatitis B infection on a frequent basis. PMID- 14556999 TI - Structure of the O-polysaccharide leads to classification of Proteus penneri 31 in Proteus serogroup O19. AB - O-polysaccharide was obtained by mild acid degradation of the lipopolysaccharide (LPS) of Proteus penneri strain 31. Sugar and methylation analyses along with NMR spectroscopic studies, including 2D 1H,1H COSY, TOCSY, ROESY, 1H,13C and 1H,31P HMQC experiments, demonstrated the following structure of the polysaccharide: [carbohydrate structure: see text] where FucNAc is 2-acetamido-2,6 dideoxygalactose and EtnP is 2-aminoethyl phosphate. The polysaccharide studied has the same carbohydrate backbone as the O-polysaccharide of Proteus vulgaris O19. Based on this finding and close serological relatedness of the LPS of the two strains, it is proposed to classify P. penneri 31 in Proteus serogroup O19 as an additional subgroup. In contrast, D-GlcNAc6PEtn and alpha-L-FucNAc-(1-->3)-D GlcNAc shared with a number of other Proteus O-polysaccharides could not provide any significant cross-reactivity of the corresponding LPS with rabbit polyclonal O-antiserum against P. penneri 31. PMID- 14557000 TI - Quantitative real-time PCR using TaqMan and SYBR Green for Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, tetQ gene and total bacteria. AB - Accurate quantification of bacterial species in dental plaque is needed for microbiological diagnosis of periodontal diseases. The present study was designed to assess the sensitivity, specificity and quantitativity of the real-time PCR using the GeneAmp Sequence Detection System with two fluorescence chemistries. TaqMan probe with reporter and quencher dye, and SYBR Green dye were used for sources of the fluorescence. Primers and probes were designed for Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia and total bacteria based on the nucleotide sequences of the respective 16S ribosomal RNA genes. Since spread of antibiotic resistance genes is one of the crucial problems in periodontal therapy, quantitative detection of tetQ gene, which confers resistance to tetracycline, was included in the examination. The detection of P. gingivalis, P. intermedia and A. actinomycetemcomitans was linear over a range of 10-10(7) cells (10-10(7) copies for tetQ gene), while the quantitative range for total bacteria was 10(2)-10(7) cells. Species-specific amplifications were observed for the three periodontal bacteria, and there was no significant difference between the TaqMan and SYBR Green chemistry in their specificity, quantitativity and sensitivity. The SYBR Green assay, which was simpler than TaqMan assay in its manipulations, was applied to the clinical plaque samples. The plaque samples were obtained from eight patients (eight periodontal pockets) before and 1 week after the local drug delivery of minocycline. Although the number of P. gingivalis, P. intermedia and A. actinomycetemcomitans markedly decreased after the antibiotic therapy in most cases, higher copy numbers of the tetQ gene were detectable. The real-time PCR demonstrated sufficient sensitivity, specificity and quantitativity to be a powerful tool for microbiological examination in periodontal disease, and the quantitative monitoring of antibiotic resistance gene accompanied with the antibiotic therapy should be included in the examination. PMID- 14557001 TI - Structure of the O-polysaccharide of Proteus penneri 28 and Proteus vulgaris O31 and classification of P. penneri 26 and 28 in Proteus serogroup O31. AB - The lipopolysaccharides (LPS) of Proteus penneri 28 and Proteus vulgaris O31 (PrK 55/57) were degraded with dilute acetic acid and structurally identical high molecular-mass O-polysaccharides were isolated by gel-permeation chromatography. Sugar analysis and nuclear magnetic resonance (NMR) spectroscopic studies showed that both polysaccharides contain D-GlcNAc, 2-acetamido-2,6-dideoxy-L-glucose (L 2-acetamido-2,6-dideoxyglucose (N-acetylquinovosamine)) and 2-acetamido-3-O-[(S) 1-carboxyethyl]-2-deoxy-D-glucose (N-acetylisomuramic acid) and have the following structure: [carbohydrate structure: see text] where (S)-1-carboxyethyl [a residue of (S)-lactic acid] (S-Lac) is an ether-linked residue of (S)-lactic acid. The O-polysaccharide studied is structurally similar to that of P. penneri 26, which differs only in the absence of S-Lac from the GlcNAc residue. Based on the O-polysaccharide structures and serological data of the LPS, it was suggested classifying these strains in one Proteus serogroup, O31, as two subgroups: O(31a), 31b for P. penneri 28 and P. vulgaris PrK 55/57 and O31a for P. penneri 26. A serological relatedness of the LPS of Proteus O(31a), 31b and P. penneri 62 was revealed and substantiated by sharing epitope O31b, which is associated with N-acetylisomuramic acid. It was suggested that a cross-reactivity of P. penneri 28 O-antiserum with the LPS of several other P. penneri strains is due to a common epitope(s) on the LPS core. PMID- 14557003 TI - Pregnancy-related changes in the longer-term management of HIV-infected women in Europe. AB - OBJECTIVES: To clarify policies regarding management of HIV-infected women, relating to pregnancy, in view of current European consensus guidelines. STUDY DESIGN: Postal questionnaire survey in 36 hospitals in 11 European countries. RESULTS: Responses were received from 22 (61%) centres, representing all 11 countries. In principle, antiretroviral therapy (ART) would be reviewed in treated women wanting to become pregnant in nearly all centres. Multidisciplinary management of infected pregnant women was routine in 17 (77%) centres, facilitating continuity of care. Approximately half the clinicians would use zidovudine monotherapy for pregnant women without indications for ART, while the remainder prescribed combination therapy. In 1998-2000, pre-eclampsia was the most prominent adverse event (22 centres) in women receiving ART, with congenital abnormalities (13 abnormalities in 6 centres) and stillbirth (5 centres) also reported. CONCLUSIONS: Policies varied, particularly regarding prophylactic ART for women without indications for treatment and did not always follow current European guidelines. PMID- 14557004 TI - Risk factors for macrosomia and its clinical consequences: a study of 350,311 pregnancies. AB - OBJECTIVES: To identify demographic risk factors for either birthweight >4kg or over the 90th centile and to quantify the obstetric risks. STUDY DESIGN: Data from 350,311 completed singleton pregnancies in the North West Thames Region between 1988 and 1997 were analysed using logistic regression. Predisposing factors and pregnancy outcome were compared by birthweight 2.5-4kg (n=259,902) and >4kg (n=36,462) and 10th-90th centile (n=279,780) and >90th centile (n=34,937). RESULTS: Macrosomia defined as birthweight >90th centile was more likely in women whose BMI >30 (kg/m(2)) (odds ratio (OR) 2.08; confidence intervals (CI) 1.99, 2.17), parity >4 (OR 2.20; CI 2.02, 2.40), age >40 (OR 1.22; CI 1.11, 1.35) and in women with pre-existing diabetes (OR 6.97; CI 5.36, 8.16) or who developed gestational diabetes (OR 2.77; CI 2.51, 3.07). Macrosomia defined by birthweight >4kg was better than birthweight >90th centile at predicting morbidity and was associated with a prolonged first and second stage of labour (OR 1.57; CI 1.51, 1.63) and (OR 2.03; CI 1.88, 2.19), respectively, an increased risk of instrumental vaginal delivery (OR 1.76; CI 1.68, 1.85), third degree perineal trauma (OR 2.73; CI 2.30, 3.23), emergency caesarean section (OR 1.84; CI 1.75, 1.93), postpartum haemorrhage (OR 2.01; CI 1.93, 2.10), Apgar score <4 (OR 1.35; CI 1.03, 1.76), and admission to the special care baby unit (OR 1.51; CI 21.38, 1.68). CONCLUSION: Macrosomia is more common in mothers who are obese, older or diabetic and is associated with significant obstetric morbidity. PMID- 14557005 TI - Which women are most affected by delivery-related changes in pelvic organ mobility? AB - OBJECTIVE: Vaginal childbirth is a risk factor for female pelvic organ prolapse and stress incontinence. Imaging studies have suggested that vaginal delivery leads to increased anterior vaginal wall mobility. The authors intended to define the relationship between antepartum pelvic organ mobility and delivery-related changes in these parameters. STUDY DESIGN: Prospective observational clinical study conducted on 200 nulliparous women recruited from antenatal clinic. Patients were assessed by translabial ultrasound in late pregnancy and 2-5 months postpartum. RESULTS: Vaginal childbirth resulted in highly significant changes to all parameters used to describe pelvic organ mobility. These alterations were more marked in those women with limited pelvic organ mobility prior to childbirth, and these negative correlations were highly significant (anterior compartment r=-0.519, P<0.0001, central compartment r=-0.539, P<0.0001, posterior compartment r=-0.604, P<0.0001). CONCLUSIONS: Women with little pelvic organ mobility before childbirth appear to be those that show most marked delivery related changes. PMID- 14557006 TI - Maternal metabolic control and perinatal outcome in women with gestational diabetes treated with regular or lispro insulin: comparison with non-diabetic pregnant women. AB - OBJECTIVE: To compare maternal glucose levels and neonatal outcome, achieved in women with gestational diabetes (GDM) receiving either regular insulin or insulin lispro, with those of a control group of non-diabetic pregnant women. STUDY DESIGN: We enrolled 49 pregnant women with GDM, randomly allocated to the treatment with either insulin lispro (n=25) or regular insulin (n=24), and 50 pregnant women with normal GCT, matched for age, parity, pre-pregnancy weight and BMI, who formed the control group. All the women were caucasian, non-obese, with a singleton pregnancy and delivered term live born infants. Women of both groups were requested to perform a blood glucose profile (consisting of nine determinations: fasting/pre-prandial, 1 and 2h post-prandial) every week from the time of diagnosis to 38 weeks (study subgroups) or every 2 weeks from 28 to 38 weeks' gestation (control group). RESULTS: Overall pre-prandial blood glucose values in diabetic women were significantly higher than those of controls; at the 1h post-prandial time point, blood glucose values of GDM women receiving insulin lispro were similar to those of controls, whereas in the regular group they were significantly higher. Overall, both the lispro and regular insulin obtained optimal metabolic control at the 2h post-prandial time point, although near normal blood glucose levels 2h after lunch could be observed only in the lispro group. There were no statistically significant differences between the groups in neonatal outcome and anthropometric characteristics; however, the rate of infants with a cranial-thoracic circumference (CC/CT) ratio between the 10th and the 25th percentile was significantly higher in the group treated with regular insulin in comparison to the lispro and control groups. CONCLUSIONS: Fasting/pre-prandial and 1h post-prandial maternal blood glucose levels in non-diabetic pregnant women fell well below the currently accepted criteria of glycemic normality in diabetic pregnancies. In women with GDM, the use of insulin lispro enabled the attainment of near-normal glucose levels at the 1h post-prandial time point and was associated with normal anthropometric characteristics; the use of regular insulin was not able to blunt the 1h peak post-prandial response to a near-normal extent and resulted in infants with a tendency toward the disproportionate growth. Insulin lispro can be regarded as a valuable option for the treatment of gestational diabetes. PMID- 14557007 TI - Social deprivation and poor access to care as risk factors for severe pre eclampsia. AB - OBJECTIVE: To estimate the associations between biomedical, social and health care factors and the occurrence of severe pre-eclampsia, eclampsia or HELLP syndrome. STUDY DESIGN: A case-control study conducted in 14 of the 15 maternity hospitals of Brussels. Cases were all 99 women who delivered in these hospitals in 1996 and who had severe pre-eclampsia, eclampsia or HELLP syndrome. Controls were 200 women without these severe maternal conditions, randomly selected among women who delivered in the same hospitals during the same period. Crude odds ratios were computed and adjusted odds ratios were derived from logistic regression. RESULTS: Indicators of social deprivation such as low educational level, poverty and illegal residence or asylum request, were strongly associated with the outcome in univariate analysis. So were African or Turkish ethnicity, obesity, chronic hypertension and primiparity. Logistic regression showed that no access to national health insurance and history of residence in another country were strongly and independently associated with the outcome (adjusted odds ratio = 4.0 (95% confidence interval 1.1, 14.0) and 3.7 (95% confidence interval 1.9, 7.3), respectively). CONCLUSIONS: The burden of pre-eclampsia is concentrated in socially disadvantaged women. Health services should be more responsive to the specific needs of these women. Low access to health care may contribute to the occurrence of severe pre-eclampsia in our setting. PMID- 14557008 TI - Pulmonary functions in pre-eclamptic women. AB - OBJECTIVE: The purpose of this study was to evaluate certain pulmonary function tests of the pre-eclamptic women in the early postpartum period. STUDY DESIGN: Forced vital capacity (FVC), forced expiratory volume in 1s (FEV(1)) and peak expiratory flow (PEF) were measured in 13 pre-eclamptic and 15 control subjects undergoing cesarean section (C/S); and 11 pre-eclamptic and 15 control subjects undergoing vaginal delivery (VD) on the postpartum third day. RESULTS: Pre eclamptic women had significantly lower FVC, FEV(1) and PEF measurements than the control women (P<0.05). When the subjects were grouped according to the mode of delivery, FVC and FEV(1) values were observed to be significantly different between the pre-eclamptic and control groups undergoing C/S (P<0.05). None of these parameters were significantly different between the pre-eclamptic and control groups who had delivered vaginally (P>0.05). CONCLUSION: These data indicate that certain pulmonary functions might be impaired in the early postpartum period in pre-eclamptic women undergoing C/S. PMID- 14557009 TI - Do prophylactic prostaglandins reduce the transfusion rate at cesarean section in high-order multiple pregnancies? AB - OBJECTIVE: Cesarean section is the more usual mode of delivery in high-order multiple pregnancy (> or =3). Excessive uterine distension increases the risk of bleeding and the need for transfusion. The aim of this study was to investigate if prophylactic use of prostaglandins at cesarean section for high-order multiple pregnancies reduces blood loss and transfusion requirement based on historic data. STUDY DESIGN: We studied a prospective series of 28 parturients with high order multiple pregnancy (group 2) who were treated, after clamping the last umbilical cord, with oxytocin (5IU intravenous then 35IU in a 24h infusion) combined with intravenous prostaglandin. A comparable retrospective series of 14 patients (group 1) had been given oxytocin alone at the same dose. Postoperative serum hemoglobin and transfusion rate as well as adverse effects were compared between the two groups. Student's t-test was used to compare continuous variables. Chi square test and Fisher exact test were used to compare categorical variables. RESULTS: The two groups were comparable for anthropometric data and duration of pregnancy. None of the patients in group 2 required red cell transfusion while 21.4% of those in group 1 required transfusion. A significant lower decrease of postoperative haemoglobin is noted in group 2 (P=0.0006). Multivariate analysis using variables significant at univariate analysis and pre eclampsia confirmed this difference. There were no adverse reactions to treatment. CONCLUSION: In our experience, prophylactic prostaglandin infusion at cesarean section in high-order multiple pregnancy is associated with a lower need for per operative red cell transfusion and a higher postoperative hemoglobin level. This observation merits confirmation in larger studies. PMID- 14557010 TI - T-helper 2-cytokine levels in women with threatened abortion. AB - OBJECTIVE: Considering that successful embryo development has been immunologically attributed to a T-helper 2 phenomenon and that threatened abortion is a very frequent but pathogenetically not well-defined clinical entity, our purpose was to investigate serum levels of the main T-helper 2-type cytokines during the evolution of this condition. STUDY DESIGN: Three T-helper 2 type cytokines (interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-13 (IL 13)) were measured by an enzyme-linked immunosorbent assay (ELISA) in serum of 12 women with threatened abortion both at hospital admission and discharge time. Fourteen women with missed abortion, 14 normal pregnant women and 14 normal non pregnant women represent study control groups. RESULTS: Serum concentrations of the selected T-helper 2-type cytokines showed no significant differences in women with threatened abortion with those of normal pregnant and non-pregnant women, whereas showed significantly lower values in women with missed abortion. CONCLUSION: Our data (a) confirm the concept that first-trimester normal pregnancy is a T-helper 2 phenomenon, (b) show that threatened abortion, when T helper 2-biased, may tend to a positive evolution of the condition, (c) display that interleukin-10, particularly, may represent a useful diagnostic and prognostic marker for predicting the normal continuance of the pregnancy in threatened abortion, (d) confirm the existence of a T-helper 2-type pattern deficiency in missed abortion, and finally (e) may open the way to new T-helper 2 biased immune therapies in case of difficult first-trimester pregnancies. PMID- 14557011 TI - A randomised comparison of side effects and patient inconvenience of two vaginal progesterone formulations used for luteal support in in vitro fertilisation cycles. AB - OBJECTIVE: To compare side effects and patient inconvenience of two vaginal progesterone (P) formulations for luteal support in in vitro fertilisation cycles. STUDY DESIGN: Sixty infertile patients at risk of developing ovarian hyperstimulation syndrome were randomised to receive either Cyclogest vaginal suppositories 400mg twice daily or Crinone 8% vaginal gel once daily for 14 days as the luteal support. On Day 6 and Day 16 after embryo transfer, they rated side effects and patient inconvenience into four grades: none, mild, moderate and severe by completing a questionnaire. RESULTS: Perineal irritation was reported by about 20% of patients in each group. Significantly more patients using Cyclogest suppositories graded inconvenience of administration, leaking out and interference with coitus as moderate or severe. CONCLUSION: There was no difference in perineal irritation after Cyclogest suppositories or Crinone 8% gel although significantly more patients found inconvenience of administration, leaking out and interference with coitus after Cyclogest. PMID- 14557012 TI - Complete down-regulation is not mandatory for good assisted reproductive treatment cycle outcomes. AB - OBJECTIVE: To determine the implications and predictive value of estradiol concentrations following pituitary down-regulation with gonadotrophin releasing hormone agonists in women undergoing controlled ovarian hyperstimulation for assisted reproductive technology. STUDY DESIGN: A total of 277 patients undergoing ovarian hyperstimulation for intracytoplasmic sperm injection (ICSI) were enrolled into the study and the patients were divided into four groups according to estradiol levels on the initial day of stimulation of which group-A consisted of the patients who had < or =25 pg/ml (n=90), group-B with levels between 26 and 50 pg/ml (n=104), group-C with levels between 51 and 75 pg/ml (n=67) and group-D with levels > or =76-90 pg/ml (n=16) and the results were compared. The primary outcome measures included ovarian response and the clinical pregnancy rates. RESULTS: The clinical pregnancy rates in groups-A, B, C and D were 33.3% (30/90), 26.0% (27/104), 35.8% (24/67), and 25.0% (4/16), respectively, and there was no statistically significant difference (P=0.482). The mean number of oocytes retrieved in groups were (9.7+/-5.8, 10.3+/-6.5, 11.0+/-6.8, and 12.1+/-6.6), respectively (P=0.453) and the fertilization rates in groups-A, B, C and D were found to be similar (75, 80, 73 and 79%, respectively; P=0.658). CONCLUSION: Complete and deep desensitization obviously seems not to be mandatory for successful stimulation in assisted reproductive technology cycles. PMID- 14557013 TI - Medico-economic approach to the management of uterine myomas: a 6-month cost effectiveness study of pelvic embolization versus vaginal hysterectomy. AB - INTRODUCTION: Uterine artery embolization is a technique that has been recently proposed for the management of uterine myomas an alternative to vaginal hysterectomy. The results provided by the first published studies demonstrate a significant decrease in symptoms in 70-95% of cases. The aim of our study was to compare the cost-effectiveness ratios for pelvic embolization and vaginal hysterectomy looked at from the hospital point of view. MATERIALS AND METHODS: Two populations of patients were randomly selected from women undergoing pelvic embolization or vaginal hysterectomy. We retrospectively measured the cost at 6 months of the two types of procedure (costs of hospitalization, drugs, and complementary examinations) as well as the success of each of them at 6 months. The cost of the techniques themselves were measured prospectively on the basis of four procedures. RESULTS: The cost-effectiveness ratios were 2320 Euros per success (mean cost 2134 Euros per effectiveness 92%) for embolization and 2789 Euros per success (mean cost 2789 Euros per effectiveness 100%) for hysterectomy. DISCUSSION: Pelvic embolization is more cost-effective than vaginal hysterectomy. The integration of the notion of quality of life with the notion of cost should permit a future study to reinforce interest in performing pelvic embolizations in the management of uterine myomas. PMID- 14557014 TI - Left:right side ratio of endometriotic implants in the pelvis. AB - OBJECTIVE: The frequency of endometriotic lesions in the right and left hemipelvis was analysed in 856 women with endometriosis. Eligible for the study were women with primary or secondary sterility, chronic pelvic pain, fibroids or benign ovarian cysts requiring laparoscopy or laparotomy consecutively observed between May 1991 and July 1992 in 23 obstetric and gynecology departments in Italy. Women with a previous diagnosis of endometriosis were excluded. A total of 3684 women entered the study. Of those, 856 had endometriosis and are considered in the present analysis. RESULTS: Five hundred and ninety four had bilateral lesions (including both ovarian and peritoneal lesions). Of the 262 women with unilateral lesions, 118 (45%, 95% confidence intervals (CI) 38-54) had the lesions in the right side of the pelvis and 144 (55%) in the left one. CONCLUSIONS: From an anatomical point of view, these findings support the transplantation therapy in the pathogenesis of endometriosis. PMID- 14557015 TI - Absence of point mutation in codons 12 and 13 of K-RAS oncogene in HPV-associated high grade dysplasia and squamous cell cervical carcinoma. AB - OBJECTIVES: Research data on the K-RAS gene mutation in carcinogenesis of the uterine cervix remain contradictory. Hence the question of whether spot mutations of the RAS genes or their excessive expression are an indispensable condition for the generation of the neoplastic phenotype of the cervical epithelial cell remains without an explicit answer. AIM OF THE STUDY: The purpose of the study was identification of point mutation in codons 12 and 13 of the first exon of K RAS gene in DNA from squamous cell cervical carcinomas, high grade dysplasias, and normal epithelium. MATERIAL AND METHODS: The study group consisted of 35 postoperative tissues from patients diagnosed with high grade dysplasia and 29 postoperative tissues from patients diagnosed with squamous cell cervical carcinoma. The control group consisted of normal cervical tissue specimens obtained from 33 patients who underwent hysterectomy due to uterine leiomyomas. Identification of point mutation in codons 12 and 13 of the first exon of K-RAS genes was performed using polymerase chain reaction (PCR)-SSCP technique. RESULTS: PCR-SSCP analysis did not reveal the presence of point mutation in codons 12 and 13 of K-RAS gene in any of the analyzed cases. PMID- 14557016 TI - Comparison of the effects of the ligation of ascending branches of bilateral arteria uterina with tourniquet method on the intra-operative and post-operative hemorrhage in abdominal myomectomy cases. AB - OBJECTIVE: To compare the effects of ligation of the ascending branches of the uterine artery and tourniquet methods intra-operative and post-operative hemorrhage during abdominal myomectomy. METHODS: Fifty-one cases with symptomatic myoma uteri were prospectively, randomly assigned to two single blind groups. Myomectomy was performed in Group 1 (n=26) by bilateral ligation of the ascending branches of the uterine artery and in Group 2 (n=25) by the intra-operative tourniquet method. Mann-Whitney U, Wilcoxon Rank and chi(2) tests were used in the comparison of the data. Spearman correlation analysis was used to determine the correlation between myoma enucleation, operation duration and blood loss (r(s), P, n). RESULTS: Operation time, number of removed myoma nuclei, (P>0.05, Mann-Whitney U test), and febrile morbidity were similar in the two groups (P>0.05, chi(2) test). Intra-operative blood loss was lower in Group 1 (P<0.05, Mann-Whitney U test). In three cases from Group 2 post-operative hemorrhage occurred, and the laparotomy and bilateral ligation of the internal iliac arteries were performed in these cases. There was a positive correlation among myoma enucleation, operation duration and blood loss (r(s)=0.7, P=0.001, n=51). CONCLUSIONS: Intra-operative blood loss is lower with bilateral ligation of the ascending branches of the uterine artery during myomectomy. The effect of ligation on hemorrhage continues in the post-operative period. PMID- 14557017 TI - Ectopic pregnancy among non-medicated and copper-containing intrauterine device users: a 10-year follow-up. AB - OBJECTIVE: To compare ectopic pregnancy (EP) rates among users of non-medicated and copper-containing IUDs. STUDY DESIGN: The number of women with non-medicated and copper devices were 3491 and 11,682, respectively. Cumulative woman-months of use (CWMU), Pearl Index (PI) and gross cumulative lifetable EP rates for 1000 women were calculated. RESULTS: The numbers of users at the end of the 10th year were 550 and 569 for non-medicated and copper-containing IUDs, respectively. CWMU for plastic and copper devices was 191,678 and 473,533, respectively, at the 169th month of maximum follow-up. The PIs were 0.8 and 1.0 calculated for 1000 users per year. The gross cumulative lifetable EP rates for 1000 women were 1.4 and 0.9 at 12 months, and 6.8 and 8.9 at the end of the 10th year, respectively, for the plastic and the copper IUDs. CONCLUSION: Although the rates were lower for the plastic IUD in the majority of the observations, there were no statistically significant differences between the two groups. PMID- 14557018 TI - New technology for diagnosis of bacterial vaginosis. AB - OBJECTIVE: To replace clinical diagnosis of bacterial vaginosis (BV) with a new and rapid analytical method based on ion mobility spectrometry (IMS). IMS is an instrumental technique for identifying compounds and determining their concentrations, based on measurement of the velocity of ions drifting through air at atmospheric pressure under the influence of an electric field. The technique is particularly sensitive to amines taking less than 2 min. STUDY DESIGN: Clinical examination of 174 samples determined 22 BV-positive and 152 BV-negative samples. IMS analyzed and recorded biogenic amine emanation mobility spectra of the 174 samples of vaginal discharge from duplicate swabs. RESULTS: IMS confirmed 21 (true positive BV) samples with 1 false negative (21/22, sensitivity=95.5%). Out of 152 samples, 150 were confirmed true BV-negative with 2 false positive samples (specificity=98.7%), PPV: 91.3%, NPV: 100%. CONCLUSIONS: The diagnostic procedure shows high accuracy and is technically simple and rapid. The trimethylamine level becomes an index of severity of the infection. PMID- 14557019 TI - Buschke-Lowenstein tumour in pregnancy. AB - During pregnancy a localised human papillomavirus (HPV) lesion may, in rare cases, develop into a Buschke-Lowenstein tumour. The choice of treatment is crucial as standard systemic treatment is teratogenic. We performed laser CO2 microsurgery because it has a low incidence of complications. PMID- 14557020 TI - Pseudocyst of the umbilical cord with mucoid degeneration of Wharton's jelly. AB - The perinatal findings of a pregnancy complicated by an umbilical cord abnormality associated with mucoid degeneration of Wharton's jelly are presented. Serial ultrasound examinations were not performed because the patient did not visit our outpatient department regularly. The umbilical cord cyst was not detected on ultrasound until delivery. PMID- 14557021 TI - Intramural pregnancy with negative maternal serum b-HCG. AB - We report an extremely rare case of intramural pregnancy with negative maternal b HCG value. The patient underwent a "myomectomy" but the pathology examination revealed chorionic villi consistent with ectopic pregnancy. Diagnosis is very difficult in cases with menorrhagia and especially when pregnancy is not suspected. PMID- 14557022 TI - Specialist life--Dr. Richard William Beard. PMID- 14557023 TI - Analysis of retropubic colpourethrosuspension results by suburethral sling with REMEEX prosthesis. PMID- 14557025 TI - 'Complementing' viral infection: mechanisms for evading innate immunity. AB - Through co-evolution with their hosts, viruses have developed a variety of immune escape and control mechanisms. In addition to strategies used to avoid the cellular and humoral immune responses, many viral families encode proteins capable of neutralizing the host's first line of defense, complement. The diversity of these complement avoidance mechanisms and proposed mechanisms by which viruses not only avoid, but also use the immune system to their advantage are discussed. PMID- 14557026 TI - Bacterial spore germination and protein mobility. AB - Fluorescence recovery after photobleaching (FRAP) of green fluorescent protein (GFP) has been used to report on protein mobility in single spores. Proteins found in dormant Bacillus spores are not mobile; however, mobility is restored when germination occurs and the core rehydrates. Spores of a cwlD mutant, in which the cortex is resistant to hydrolysis, are able to complete the earliest stages of germination in response to a specific germinant stimulus; in these circumstances, the protein in the spore remains immobile. Therefore, the earliest stages of spore germination, including loss of resistance to extreme heat and the complete release of the spore component dipicolinic acid, are achieved without the restoration of protein mobility. PMID- 14557027 TI - Searching for keys under the devil's lamppost. PMID- 14557028 TI - What is driving the acquisition of mutS and rpoS polymorphisms in Escherichia coli? AB - Pathogenic and commensal Escherichia coli isolates frequently contain defective alleles of the mutS and rpoS genes, located in a highly polymorphic segment of the chromosome. The environments leading to enrichment of rpoS mutations and the selective advantages of these mutants are becoming apparent. Unexpectedly, rpoS defects occur because of a basic design limitation in cellular regulation. Antagonistic pleiotropy results from the futile competition between different sigma factors associated with the RNA polymerase, and drives the elimination of RpoS (or sigma(S)) in environments requiring high levels of transcription that is dependent on RpoD (or sigma(D) or sigma(70)). Nutrient-limited environments provide an ideal breeding ground for rpoS mutations. By contrast, in other settings, increased stress resistance selects for restoration of rpoS function. Hence extensive polymorphism in the mutS-rpoS region is postulated to result from cycling between environments in which the functional or non-functional genes provide distinct fitness advantages. PMID- 14557029 TI - Oomycetes and fungi: similar weaponry to attack plants. AB - Fungi and Oomycetes are the two most important groups of eukaryotic plant pathogens. Fungi form a separate kingdom and are evolutionarily related to animals. Oomycetes are classified in the kingdom Protoctista and are related to heterokont, biflagellate, golden-brown algae. Fundamental differences in physiology, biochemistry and genetics between fungi and Oomycetes have been described previously. These differences are also reflected in the large variations observed in sensitivity to conventional fungicides. Recently, more pronounced differences have been revealed by genomics approaches. However, in this review we compare the mode of colonization of the two taxonomically distinct groups and show that their strategies have much in common. PMID- 14557030 TI - Progress in functional genomics approaches to antifungal drug target discovery. AB - Antifungal drug discovery is starting to benefit from the enormous advances in the genomics field, which have occurred in the past decade. As traditional drug screening on existing targets is not delivering the long-awaited potent antifungals, efforts to use novel genetics and genomics-based strategies to aid in the discovery of novel drug targets are gaining increased importance. The current paradigm in antifungal drug target discovery focuses on basically two main classes of targets to evaluate: genes essential for viability and virulence or pathogenicity factors. Here we report on recent advances in genetics and genomics-based technologies that will allow us not only to identify and validate novel fungal drug targets, but hopefully in the longer run also to discover potent novel therapeutic agents. Fungal pathogens have typically presented significant obstacles when subjected to genetics, but the creativity of scientists in the anti-infectives field and the cross-talk with scientists in other areas is now yielding exciting new tools and technologies to tackle the problem of finding potent, specific and non-toxic antifungal therapeutics. PMID- 14557031 TI - Multi-locus sequence typing: a tool for global epidemiology. AB - The characterization of pathogenic isolates plays a pivotal role in the epidemiology of infectious diseases, generating the information necessary for identifying, tracking, and intervening against disease outbreaks. In 1998 multi locus sequence typing (MLST) was proposed as a nucleotide sequence-based approach that could be applied to many bacterial pathogens. It combined developments in high-throughput sequencing and bioinformatics with established population genetics techniques to provide a portable, reproducible, and scalable typing system that reflected the population and evolutionary biology of bacterial pathogens. MLST schemes have been developed for a variety of procaryotic and eucaryotic pathogens and the data generated have contributed to both epidemiological surveillance and fundamental studies of pathogen biology. PMID- 14557032 TI - Current diagnosis of histoplasmosis. AB - Histoplasmosis is a common infection endemic in many regions of America, Asia, India and Africa, with sporadic cases also occurring throughout the world. Although excellent laboratory methods for diagnosis are available, there are deficiencies that must be met by continued research. Clinicians and laboratory directors must be familiar with the uses and limitations of a battery of serologic and mycological tests to accurately diagnose histoplasmosis. Research is needed to reduce false-negative and false-positive results, and to improve the identification of the organism in tissues. Approaches to the diagnosis of histoplasmosis and areas that require further research will be reviewed. PMID- 14557033 TI - Simulating transport of non-Chernobyl (137)Cs and (90)Sr in the North Atlantic Arctic region. AB - The spatial and temporal distributions of the anthropogenic radionuclides (137)Cs and (90)Sr, originating from nuclear bomb testing and the Sellafield reprocessing plants in the Irish Sea, are simulated using a global version of the Miami Isopycnic Coordinate Ocean Model (MICOM). The physical model is forced with daily atmospheric re-analysed fields for the period 1950 to present. Comparison of temporal evolution of observed and simulated concentrations of (137)Cs have been conducted for the regions east of Scotland, west of central Norway and at the entrance of the Barents Sea. It follows that the radionuclides from the Sellafield discharge reach the Barents Sea region after 4-5 years, in accordance with observations. The simulation provides a detailed distribution and evolution of the radionuclides over the integration time. For the Atlantic waters off the coast of Norway and in the southern Barents Sea, the atmospheric fallout dominates over the Sellafield release up to the mid 1960s and from the early 1990s, whereas Sellafield is the main source for the two radionuclides in the 1970s and 1980s. It is furthermore argued that model systems like the one presented here can be used for future prediction of radioactive contaminations in the Nordic Seas and the Arctic Ocean, for instance under various global warming scenarios. PMID- 14557034 TI - Radon-222 signatures of natural ventilation regimes in an underground quarry. AB - Radon-222 activity concentration has been monitored since 1999 in an underground limestone quarry located in Vincennes, near Paris, France. It is homogeneous in summer, with an average value of 1700 Bq m(-3), and varies from 730 to 1450 Bq m( 3) in winter, indicating natural ventilation with a rate ranging from 0.5 to 2.4 x 10(-6) s(-1) (0.04-0.22 day(-1)). This hypothesis is supported by measurements in the vertical access pit where, in winter, a turbulent air current produces a stable radon profile, smoothly decreasing from 700 Bq m(-3) at 20 m depth to 300 Bq m(-3) at surface. In summer, a thermal stratification is maintained in the pit, but the radon-222 concentration jumps repeatedly between 100 and 2000 Bq m( 3). These jumps are due to atmospheric pressure pumping, which induces ventilation in the quarry at a rate of about 0.1 x 10(-6) s(-1) (0.009 day(-1)). Radon-222 monitoring thus provides a dynamical characterisation of ventilation regimes, which is important for the assessment of the long-term evolution of underground systems. PMID- 14557035 TI - Polonium-210 budget in cigarettes. AB - Due to the relatively high activity concentrations of (210)Po and (210)Pb that are found in tobacco and its products, cigarette smoking highly increases the internal intake of both radionuclides and their concentrations in the lung tissues. That might contribute significantly to an increase in the internal radiation dose and in the number of instances of lung cancer observed among smokers. Samples of most frequently smoked fine and popular brands of cigarettes were collected from those available on the Egyptian market. (210)Po activity concentrations were measured by alpha spectrometry, using surface barrier detectors, following the radiochemical separation of polonium. Samples of fresh tobacco, wrapping paper, fresh filters, ash and post-smoking filters were spiked with (208)Po for chemical recovery calculation. The samples were dissolved using mineral acids (HNO(3), HCl and HF). Polonium was spontaneously plated-out on stainless steel disks from diluted HCl solution. The (210)Po activity concentration in smoke was estimated on the basis of its activity in fresh tobacco and wrapping paper, fresh filter, ash and post-smoking filters. The percentages of (210)Po activity concentrations that were recovered from the cigarette tobacco to ash, post-smoking filters, and smokes were assessed. The results of this work indicate that the average (range) activity concentration of (210)Po in cigarette tobacco was 16.6 (9.7-22.5) mBq/cigarette. The average percentages of (210)Po content in fresh tobacco plus wrapping paper that were recovered by post-smoking filters, ash and smoke were 4.6, 20.7 and 74.7, respectively. Cigarette smokers, who are smoking one pack (20 cigarettes) per day, are inhaling on average 123 mBq/d of (210)Po and (210)Pb each. The annual effective doses were calculated on the basis of (210)Po and (210)Pb intake with the cigarette smoke. The mean values of the annual effective dose for smokers (one pack per day) were estimated to be 193 and 251 microSv from (210)Po and (210)Pb, respectively. PMID- 14557036 TI - Global fallout (137)Cs accumulation and vertical migration in selected soils from South Patagonia. AB - The spatial distribution and vertical migration of global fallout (137)Cs were studied in soils from South Patagonia at the austral region of South America in semi-natural and natural environments located between 50-54 degrees S and 68-74 degrees W. The (137)Cs areal activity density varied from 222 to 858 Bq m(-2), and was found to be significantly positively correlated (p<0.001) with the mean annual precipitation rate. The fraction of the total activity density observed in steppe grass varied from <0.03% to 0.12% (median <0.07%) and is considerably lower than the results obtained at the South Shetland Islands (median 8%) and in other temperate environments in south-central Chile (median 0.2%). The median of the convection velocity v(s) of (137)Cs in the soil in such polar isotundra climate has been determined to be 0.056 cm y(-1). This value is higher than v(s) determined under polar climate (-0.012 cm y(-1)) and is near to the upper limit of v(s)-values determined in temperate environments from Chile (0.019 cm y(-1)). The median value of the diffusion coefficient D(s) (0.048 cm(2) y(-1)) is similar to D(s) observed in an Antarctic region (0.043 cm(2) y(-1)) and lower than D(s) in temperate regions of Chile (1.24 cm(2) y(-1)). About 35 years after the highest depositions, (137)Cs had penetrated to a depth of 6-14 cm in the Patagonian soils and can be expected to remain in the rooting zone of grass for many decades. Nevertheless, because of its low transfer to steppe grass observed at this region, the radioecological sensitivity of this ecosystem with respect to fallout radiocesium seems to be lower than in other polar regions. PMID- 14557037 TI - Thorium and uranium contents in human urine: influence of age and residential area. AB - Inductively coupled plasma mass spectrometry (ICP-MS) has been used for the determination of (232)Th and (238)U in urine of unexposed Jordanian subjects living in six cities. The range of (232)Th excretion in all subjects was found to be 1.4-640 microBq d(-1) with an average of 34.8 microBq d(-1) (geometric mean 15.8 microBq d(-1)). Results showed no statistically significant correlation with age and residential area. The average value obtained is in agreement with levels considered normal in some recent publications. The average value of (238)U in all samples was found to be 3955 microBq d(-1) (geometric mean 1107 microBq d(-1)), which is higher than reported figures from Germany and India, but in agreement with those figures given in ICRP publication, number 23. The mean values of the different groups were found to be proportional to age up to 60 years. A noticeable drop is observed for subjects greater than 60 years old. PMID- 14557038 TI - Soil-water distribution coefficients and plant transfer factors for (134)Cs, (85)Sr and (65)Zn under field conditions in tropical Australia. AB - Measurements of soil-to-plant transfer of (134)Cs, (85)Sr and (65)Zn from two tropical red earth soils ('Blain' and 'Tippera') to sorghum and mung crops have been undertaken in the north of Australia. The aim of the study was to identify factors that control bioaccumulation of these radionuclides in tropical regions, for which few previous data are available. Batch sorption experiments were conducted to determine the distribution coefficient (K(d)) of the selected radionuclides at pH values similar to natural pH values, which ranged from about 5.5 to 6.7. In addition, K(d) values were obtained at one pH unit above and below the soil-water equilibrium pH values to determine the effect of pH. The adsorption of Cs showed no pH dependence, but the K(d) values for the Tippera soils (2300-4100 ml/g) exceeded those for the Blain soils (800-1200 ml/g) at equilibrium pH. This was related to the greater clay content of the Tippera soil. Both Sr and Zn were more strongly adsorbed at higher pH values, but the K(d) values showed less dependence on the soil type. Strontium K(d)s were 30-60 ml/g whilst Zn ranged from 160 to 1630 ml/g for the two soils at equilibrium pH. With the possible exception of Sr, there was no evidence for downward movement of radionuclides through the soils during the course of the growing season. There was some evidence of surface movement of labelled soil particles. Soil-to-plant transfer factors varied slightly between the soils. The average results for sorghum were 0.1-0.3 g/g for Cs, 0.4-0.8 g/g for Sr and 18-26 g/g for Zn (dry weight) with the initial values relating to Blain and the following values to Tippera. Similar values were observed for the mung bean samples. The transfer factors for Cs and Sr were not substantially different from the typical values observed in temperate studies. However, Zn transfer factors for plants grown on both these tropical soils were greater than for soils in temperate climates (by more than an order of magnitude). This may be related to trace nutrient deficiency and/or the growth of fungal populations in these soils. The results indicate that transfer factors depend on climatic region together with soil type and chemistry and underline the value of specific bioaccumulation data for radionuclides in tropical soils. PMID- 14557039 TI - Waste disposal in plants: where and how? PMID- 14557040 TI - Role of death in providing lifeline to plants. AB - As the major transporters and distributors of water and minerals, xylem vessels and tracheids are the lifeline of plants. Interestingly, the building blocks of these water pipes are dead tracheary elements and vessel elements that have the process of cell death integrated into their differentiation programme. Using the Zinnia in vitro model system for xylogenesis, a key nuclease that is responsible for nuclear degradation during the terminal stages of tracheary element differentiation has been identified recently. PMID- 14557041 TI - Plant iNOS: conquest of the Holy Grail. PMID- 14557042 TI - Alternative splicing and proteome diversity in plants: the tip of the iceberg has just emerged. AB - Alternative splicing has recently emerged as one of the most significant generators of functional complexity in several relatively well-studied animal genomes, but little is known about the extent of this phenomenon in higher plants. However, recent computational and experimental studies discussed here suggest that alternative splicing probably plays a far more significant role in the generation of proteome diversity in plants than was previously thought. PMID- 14557043 TI - CAF-1 and MSI1-related proteins: linking nucleosome assembly with epigenetics. PMID- 14557044 TI - And then there were many: MADS goes genomic. AB - During the past decade, MADS-box genes have become known as key regulators in both reproductive and vegetative plant development. Traditional genetics and functional genomics tools are now available to elucidate the expression and function of this complex gene family on a much larger scale. Moreover, comparative analysis of the MADS-box genes in diverse flowering and non-flowering plants, boosted by bioinformatics, contributes to our understanding of how this important gene family has expanded during the evolution of land plants. Therefore, the recent advances in comparative and functional genomics should enable researchers to identify the full range of MADS-box gene functions, which should help us significantly in developing a better understanding of plant development and evolution. PMID- 14557045 TI - Forward genetics and map-based cloning approaches. AB - Whereas reverse genetics strategies seek to identify and select mutations in a known sequence, forward genetics requires the cloning of sequences underlying a particular mutant phenotype. Map-based cloning is tedious, hampering the quick identification of candidate genes. With the unprecedented progress in the sequencing of whole genomes, and perhaps even more with the development of saturating marker technologies, map-based cloning can now be performed so efficiently that, at least for some plant model systems, it has become feasible to identify some candidate genes within a few months. This, in turn, will boost the use of forward genetics approaches, as applied (for example) to isolating genes involved in natural variation and genes causing phenotypic mutations as derived from (second-site) mutagenesis screens. PMID- 14557046 TI - A role for the ubiquitin-26S-proteasome pathway in gibberellin signaling. AB - The gibberellin (GA) signaling pathway, like auxin and jasmonate signaling, uses the ubiquitin-proteasome pathway to control expression through protein degradation. A conserved F-box protein of an SCF E3 ubiquitin ligase is a positive regulator of GA signaling in Arabidopsis and rice. GA apparently stimulates stem elongation by causing this SCF complex to regulate negatively a family of negative regulators of GA response (the DELLA family of putative transcription factors). The DELLA family members AtRGA or (Repressor of ga1-3) and OsSLR1 (SLENDER RICE1) proteins both appear to be subject to GA-induced proteolysis. The need to have the F-box genes AtSLY1 and OsGID2 for this proteolysis suggests that GA causes proteolysis of AtRGA/OsSLR1 via the SCF(AtSLY1/OsGID2) ubiquitin ligase. PMID- 14557047 TI - Mechanotransduction in gravisensing cells. AB - Plant organs can re-orientate themselves with respect to gravity. Gravisensing cells (statocytes) contain movable amyloplasts whose potential energy is apparently used to activate calcium channels by exerting tension on the actin network and/or pressure on the cytoskeleton elements lining the plasma membrane. The chain of events that follows remains to be further analysed but includes transient pH changes in the cytosol and sustained pH changes in the cell wall. Transduction ends with relocation of the auxin efflux carriers responsible for the lateral transport of auxin, which reorients the root tip in the direction of gravity. Many questions remain to be solved but recent studies now herald a better understanding of the molecular events involved in gravisensing. PMID- 14557049 TI - ESCVS 2003. Abstracts of the 52nd International Congress of the European Society for Cardiovascular Surgery. November 7-10, 2003. Istanbul, Turkey. PMID- 14557048 TI - Calcium/calmodulin-mediated signal network in plants. AB - Various extracellular stimuli elicit specific calcium signatures that can be recognized by different calcium sensors. Calmodulin, the predominant calcium receptor, is one of the best-characterized calcium sensors in eukaryotes. In recent years, completion of the Arabidopsis genome project and advances in functional genomics have helped to identify and characterize numerous calmodulin binding proteins in plants. There are some similarities in Ca(2+)/calmodulin mediated signaling in plants and animals. However, plants possess multiple calmodulin genes and many calmodulin target proteins, including unique protein kinases and transcription factors. Some of these proteins are likely to act as "hubs" during calcium signal transduction. Hence, a better understanding of the function of these calmodulin target proteins should help in deciphering the Ca(2+)/calmodulin-mediated signal network and its role in plant growth, development and response to environmental stimuli. PMID- 14557050 TI - Association of PADI4 and rheumatoid arthritis: a successful multidisciplinary approach. AB - The identification of functionally relevant polymorphisms of peptidylarginine deiminase 4, an enzyme that catalyzes the post-translational citrullination of proteins, as a rheumatoid arthritis gene is one of the most convincing success stories of complex disease gene mapping to date. In addition to an extensive single nucleotide polymorphism-based association study in a Japanese cohort, a range of techniques have been used to validate this finding. PMID- 14557051 TI - Interaction makes the heart grow stronger. AB - A novel interaction between GATA4 and TBX5 could explain phenotypic similarities (atrial septal defects) between patients with mutations in GATA4 or TBX5. The cardiac transcription factor GATA4 has not previously been implicated in a human disorder but a recent paper by Garg et al. provides evidence of mutations in GATA4 that cause atrial septal defects. Mutations in TBX5 have already been shown to cause similar atrial septal defects in Holt-Oram syndrome. PMID- 14557052 TI - Death and the retrovirus. AB - A flurry of new papers has shown that HIV reverse transcription is vulnerable to G-->A hypermutation. Apparently, cytidine bases in nascent DNA synthesis are lethally edited by the host cell molecule apolipoprotein B editing complex protein (APOBEC) 3G. This death mechanism is circumvented by the HIV viral infectivity factor protein, which prevents APOBEC3G from entering the virion. PMID- 14557053 TI - Huntington's disease: a synaptopathy? AB - Huntington's disease (HD) is caused by a polyglutamine expansion in the protein huntingtin. In its terminal stage, HD is characterized by widespread neuronal death in the neocortex and the striatum. Classically, this neuronal death has been thought to underlie most of the symptoms of the disease. Accumulating evidence suggests, however, that cellular dysfunction is important in the pathogenesis of HD. We propose that specific impairment of the exocytosis and endocytosis machinery contributes to the development of HD. We also suggest that abnormal synaptic transmission underlies the early symptoms of HD and can contribute to the triggering of cell death in later stages of the disease. PMID- 14557055 TI - In vivo gene therapy for diabetes mellitus. AB - Gene therapy has been hyped as a possible 'cure' for diabetes mellitus in the near future ever since insulin was first cloned and expressed in cultured cells in the late 1970s. In the past decade, however, the bar for gene therapy for diabetes has been raised because of recent advances in the clinical management of diabetes. Although current treatment modalities fall far short of a cure, they produce greatly improved, if imperfect, glycemic control. In this context, we review the latest advances in in vivo gene therapy and conclude that the most widely applied strategy of insulin gene transfer does not measure up to the existing treatment options, whereas the recently proved concept of induced islet neogenesis has the potential of bettering the currently available therapy. Much work remains to be done, however, before this regimen can be taken from the bench to the bedside. PMID- 14557054 TI - Transcriptionally targeted gene therapy to detect and treat cancer. AB - The greatest challenge in cancer treatment is to achieve the highest levels of specificity and efficacy. Cancer gene therapy could be designed specifically to express therapeutic genes to induce cancer cell destruction. Cancer-specific promoters are useful tools to accomplish targeted expression; however, high levels of gene expression are needed to achieve therapeutic efficacy. Incorporating an imaging reporter gene in tandem with the therapeutic gene will allow tangible proof of principle that gene expression occurs at the correct location and at a sufficient level. Gene-based imaging can advance cancer detection and diagnosis. By combining the cancer-targeted imaging and therapeutic strategies, the exciting prospect of a 'one-two punch' to find hidden, disseminated cancer cells and destroy them simultaneously can potentially be realized. PMID- 14557056 TI - Stem cell therapy for ischemic heart disease. AB - Recent experimental and clinical observations have suggested that cell transplantation could be of therapeutic value for the treatment of heart disease. This approach was based on the idea that transplanted donor cardiomyocytes would integrate with the host myocardium and thereby directly contribute to cardiac function. Surprisingly, the observation that non-cardiomyogenic cells could also improve cardiac function indicates that functional integration of donor cells might not be required to achieve a beneficial effect. More recently, several observations have suggested the presence of a greater than anticipated developmental repertoire in adult-derived stem cells, which, if further validated, would offer unprecedented opportunities for the restoration of cardiac function in diseased hearts. Here, we discuss current issues regarding the potential use of stem cell transplantation for the treatment of ischemic heart disease. PMID- 14557057 TI - Cholesterol-receptor-mediated genomics in health and disease. AB - Cross-talk between cell-surface receptor C(k) and intracellular receptors (liver X receptor-alpha and peroxisome-proliferator-activated receptors) controls a set of crucial genes that maintain a finely orchestrated balance between various cellular processes involved in cell growth, differentiation, apoptosis, cholesterol homeostasis and inflammation. Abnormal cross-talk of these receptors can lead to several human diseases, particularly atherosclerosis, cancer and autoimmune diseases. As our understanding of the complex signaling events that link these receptors to human health improves, we are beginning to appreciate the enormous potential of the proposed cross-talk model of cholesterol receptors in the prevention and/or treatment of diseases. PMID- 14557058 TI - Significance of immune response to enzyme-replacement therapy for patients with a lysosomal storage disorder. AB - Lysosomal storage disorders are collectively important because they cause significant morbidity and mortality. Patients can present with severe symptoms that include somatic tissue and bone pathology, developmental delay and neurological impairment. Enzyme-replacement therapy has been developed as a treatment strategy for patients with a lysosomal storage disorder, and for many of these disorders this treatment is either in clinical trial or clinical practice. One major complication arising from enzyme infusion into patients with a lysosomal storage disorder is an immune response to the replacement protein. From clinical trials, it is clear that there is considerable variability in the level of immune response to enzyme-replacement therapy, dependent upon the replacement protein being infused and the individual patient. Hypersensitivity reactions, neutralizing antibodies to the replacement protein and altered enzyme targeting or turnover are potential concerns for patients exhibiting an immune response to enzyme-replacement therapy. The relative occurrence and significance of these issues have been appraised. PMID- 14557059 TI - Roles for BTAF1 and Mot1p in dynamics of TATA-binding protein and regulation of RNA polymerase II transcription. AB - Regulation of RNA polymerase II (pol II) transcription is a highly dynamic process requiring the coordinated interaction of an array of regulatory proteins. Central to this process is the TATA-binding protein (TBP), the key component of the multiprotein complex TFIID. Interaction of TBP with core promoters nucleates the assembly of the preinitiation complex and subsequent recruitment of pol II. Despite recent advances in our understanding of the dynamic nature of the pol II transcription apparatus, the dynamics of TBP function on pol II promoters has remained largely unexplored. Human BTAF1 (TAF(II)170/TAF-172) and its yeast ortholog, Mot1p, are evolutionarily conserved members of the SNF2-like family of ATPase proteins. Genetic identification of Mot1p as a repressor of pol II transcription was supported by findings that Mot1p and BTAF1 could dissociate TBP from TATA DNA complexes using the energy of ATP hydrolysis. Recent data have revealed new aspects of BTAF1 and Mot1p as positive regulators of TBP function in the pol II system and have described new observations relating to their molecular mechanism of action. We review these data in the context of previous findings with particular attention paid to how human BTAF1 and Mot1p may dynamically regulate TBP function on pol II promoters in cells. PMID- 14557060 TI - Structure of the micronuclear alpha-tubulin gene in the phyllopharyngean ciliate Chilodonella uncinata: implications for the evolution of chromosomal processing. AB - Ciliates are a group of microbial eukaryotes defined by the presence of dimorphic nuclei-each cell contains both a transcriptionally active macronucleus and a germline micronucleus. During the development of the macronucleus, germline chromosomes are rearranged through extensive fragmentation, removal of internally excised sequences (IESs) and DNA amplification. We have characterized three IESs in the gene that encodes alpha-tubulin in the phyllopharyngean ciliate Chilodonella uncinata. The IESs are located within the coding domain, range in size from 81 to 107 bp, and are flanked by direct repeats that vary in length from 6 to 8 bp. All three IESs are moderately AT-rich and each contains two copies of a conserved sequence motif. These data provide evidence for the existence of IESs in phyllopharyngean ciliates and suggest that IES processing in C. uncinata may rely on a novel cis-acting sequence. Comparisons of the IESs in C. uncinata with those of 'model' ciliates-Paramecium, Tetrahymena, Euplotes, Oxytricha and Stylonychia-reveal considerable variation in chromosomal processing among ciliates. PMID- 14557061 TI - Complete DNA sequences of the botulinum neurotoxin complex of Clostridium botulinum type A-Hall (Allergan) strain. AB - BOTOX is manufactured with the purified native 900-kDa type A neurotoxin complex from Clostridium botulinum type A-Hall (Allergan) strain. This complex is composed of the botulinum neurotoxin (BoNT) and several toxin associated proteins known as the hemagglutinins (HAs) and the non-toxic non-hemagglutinin protein (NTNH). We describe here the complete gene sequences of the BoNT complex of type A-Hall (Allergan) strain. Using a polymerase chain reaction-based approach, we sequenced six open reading frames (ORFs) encoding BoNT (1296 amino acids), the toxin-associated proteins: HA70, 625 aa; HA17, 147 aa; HA34, 291 aa; NTNH, 1193 aa; and the regulatory component botR/OrfX, 178 aa. Comparative alignments of the amino acid sequence of BoNT/A shows a 98-100% sequence identity among different strains of the type A, except for the Kyoto-F strain (90%), whereas the sequence identity between BoNT/A and other toxin serotypes is only 30.4-39.1%. Similar to the neurotoxin, the toxin-associated proteins and botR from type A-Hall strain also share more than 95% identity to the homologous proteins found in type A NCTC2916 strain. Among all the toxin associated proteins, NTNHs and HA70s are the most conserved with 65-87% identity across different serotypes. On the other hand, HA34s, present only in serotypes A-D, show greater diversity than all other toxin-associated proteins; HA34/A has 90% identity to HA34/B and only approximately 35% identity to HA34/C and HA34/D. Relatively higher sequence identity ( approximately 60%) is seen in HA17 and botR of Hall A when compared to their counterparts in serotypes C or D. Of all proteins within the toxin complex, NTNH and HA70 have the highest degree of conservation across serotypes and this may underscore a critical role for these proteins in the formation of the complexes. Physiologically, different duration of action in different serotypes may be due to different modifications of toxins by neuronal enzymes, which lead to different compartmentalization of different toxins. Computer-assisted motif analysis reveals that toxins contain several potential sites for phosphorylation by casein kinase II, protein kinase C, tyrosine kinases, glycogen synthase kinase 3, cGMP dependent protein kinase (PKG) that are well conserved. The reported sequence information for type A-Hall strain will potentially facilitate elucidation of the toxin interactions with the nontoxin proteins in the complex. PMID- 14557062 TI - Egr-1 target genes in human endothelial cells identified by microarray analysis. AB - Early growth response factor 1 (Egr-1) is a key transcriptional factor to mediate gene expression after vascular injury. To better understand the role of Egr-1 in vasculature, we globally profiled Egr-1 target genes in human endothelial cells using adenoviral gene transfer and Affymetrix oligonucleotide-based microarray technology. More than 300 genes regulated by >/=3-fold with Egr-1 overexpression were identified and, partially, confirmed by Northern and Western blotting, including genes for transcriptional regulators, signaling proteins, cell cycle regulatory proteins, growth factors, and cytokines. Among them, thymus-expressed chemokine (TECK) and IP-30 were dramatically induced by Egr-1, but TNFalpha related apoptosis inducing ligand (TRAIL) was significantly repressed by Egr-1, suggesting that Egr-1 is a key mediator of inflammation and apoptosis in vascular cells. These data provide novel Egr-1 target genes and contribute to the understanding of the role of Egr-1 in vasculature. PMID- 14557063 TI - Endonuclease genes up-regulated in tissues undergoing programmed cell death are expressed during male gametogenesis in barley. AB - In the process of programmed cell death (PCD), a key role has been attributed to endonucleases capable to cleave nuclear DNA at internucleosomal sites. In barley (Hordeum vulgare L.), two such nucleases (Bnuc1 and BEN1) were individually identified in unrelated tissues. In the present work, we demonstrate that their genes are also expressed in immature anthers at different stages of pollen development. Further experiments carried out on RNA extracted from immature barley anthers led to discover a novel endonuclease gene, namely Bnuc2 (AJ311603 in the EMBL/GenBank/DDBJ databases), eventually found up-regulated at the tetrad stage. The protein encoded was found to conserve large sequence portions of Bnuc1 and BEN1 endonucleases, including the domain regions involved in secretion and DNA/RNA binding. A survey conducted on barley EST libraries showed that Bnuc2 and BEN1 mRNAs are jointly present also in the transcriptome of 20 DAP spike and that other endonuclease ESTs are co-expressed with Bnuc1 or BEN1 in tissues where PCD has been recorded. Therefore, it can be concluded that during the PCD process, a set of S1-type endonucleases is synthesised regardless of the tissue considered. PMID- 14557064 TI - Mutational library analysis of selected amino acids in the receptor binding domain of envelope of Akv murine leukemia virus by conditionally replication competent bicistronic vectors. AB - The envelope protein of retroviruses is responsible for viral entry into host cells. Here, we describe a mutational library approach to dissect functional domains of the envelope protein involving a retroviral vector, which expresses both the envelope protein of Akv murine leukemia virus (MLV) and the neomycin phosphotransferase II (Neo) selection marker from the same transcript. Envelope expression was achieved by inserting an internal ribosome entry site (IRES) between the neo and the env genes. We found the structure of the linker between the IRES element and env to be critical for sufficient envelope expression. This vector functions as a replication competent mini-virus in a culture of NIH 3T3 derived semi-packaging cells that express the viral Gag and Pol proteins. Titers comparable to those of wild type virus were achieved by this system. To test this vector system, we created a random mutational library of Arg 85 and Asp 86 in the first variable region of Akv envelope protein. Homologous amino acids to Asp 86 in Moloney and Friend murine leukemia viruses are thought to be directly involved in receptor binding. Subsequent selection of mutants capable of infecting murine NIH 3T3 cells indicated that the wild type aspartic acid or another hydrophilic residue at position 86 is an important determinant for envelope function. PMID- 14557065 TI - Mini-lambda: a tractable system for chromosome and BAC engineering. AB - The bacteriophage lambda (lambda) recombination system Red has been used for engineering large DNA fragments cloned into P1 and bacterial artificial chromosomes (BAC or PAC) vectors. So far, this recombination system has been utilized by transferring the BAC or PAC clones into bacterial cells that harbor a defective lambda prophage. Here we describe the generation of a mini-lambda DNA that can provide the Red recombination functions and can be easily introduced by electroporation into any E. coli strain, including the DH10B-carrying BACs or PACs. The mini-lambda DNA integrates into the bacterial chromosome as a defective prophage. In addition, since it retains attachment sites, it can be excised out to cure the cells of the phage DNA. We describe here the use of the mini-lambda recombination system for BAC modification by introducing a selectable marker into the vector sequence of a BAC clone. In addition, using the mini-lambda, we create a single missense mutation in the human BRCA2 gene cloned in a BAC without the use of any selectable marker. The ability to generate recombinants very efficiently demonstrates the usefulness of the mini-lambda as a very simple mobile system for in vivo genome engineering by homologous recombination, a process named recombineering. PMID- 14557066 TI - Isolation and characterization of a novel human thioredoxin-like gene hTLP19 encoding a secretory protein. AB - A novel human gene, named as hTLP19, was isolated and characterized as secretory protein by combining bioinformatics tools with experiments. The hTLP19 encodes 172 amino acid residues with signal peptide in its N-terminal and a thioredoxin (Trx) domain that is homologous with some genes in Mus musculus, Xenopus laevis, etc. Moreover, the result from insulin reduction assay indicated that the hTLP19 protein has Trx enzymatic activity. By comparing full-length cDNA with human genomic DNA, the hTLP19 gene might have seven coding exons spanning 35 kb of genomic DNA on the human chromosome 1p32.3. Northern blot analysis showed that human hTLP19 is expressed widely in many tissues with 1.6 kb band and extra 1.2 kb band in placenta. Subcellular localization and immunoblotting assays indicated that hTLP19 might be secreted out of cell through trans-Golgi network (TGN). PMID- 14557067 TI - Expression modes of interferon-alpha inducible genes in sensitive and resistant human melanoma cells stimulated with regular and pegylated interferon-alpha. AB - Interferon-alpha with its antiproliferative activity is widely used for the treatment of viral infections and tumor therapy such as melanoma. Naturally occurring resistance to recombinant interferon alpha-2a (IFN-alpha) and severe side effects limit the therapeutic efficacy. Understanding of the molecular mechanisms involved in unresponsiveness may therefore lead to the development of novel formulations that overcome resistance. Here, we have applied oligonucleotide DNA microarrays with probe sets for about 11,400 human transcripts to study the expression of interferon-alpha inducible genes in a sensitive and resistant melanoma cell line over multiple time points and two interferon formulations. We identified two major groups of genes with termed interferon primary response genes (IPRGs) or interferon secondary response genes (ISRGs). IPRGs are upregulated early after interferon stimulation in both the sensitive and the resistant line and they contain IREs in the noncoding regulatory region. In contrast, ISRG expression occurs preferentially in the sensitive line ME15 at late time points, and this group of genes lacks typically IREs. In addition to these two major interferon response gene classes, we identified a relatively small number of genes with complex kinetic expression modes. In addition, we show for the first time that regular and pegylated recombinant interferons are equally potent activators of interferon (IFN) gene expression. Finally, we propose that the ISRGs are activated downstream of the primary response genes by a molecule or pathway, which awaits identification, and interferon inducible gene expression is thus more complicated than previously thought. PMID- 14557068 TI - Human SLITRK family genes: genomic organization and expression profiling in normal brain and brain tumor tissue. AB - Slitrk family proteins are characterized as integral membrane proteins that have two leucine-rich repeat (LRR) domains and a carboxy-terminal domain that is partially similar to trk neurotrophin receptor proteins. The LRR domains are similar to those of slit proteins. In a previous study, we showed that mouse Slitrk genes are expressed predominantly in neural tissue and have neurite modulating activity in cultured neuronal cells. Their expression profiles as well as their functions vary among the family members. In this paper, we characterized the human SLITRK1, SLITRK2, SLITRK3, SLITRK4, SLITRK5, and SLITRK6 genes. The six genes are located in three clusters, on 3q, 13q, and Xq, respectively. Their expression was detected mainly in the brain, but the expression profile of each SLITRK was unique. SLITRK expression was also investigated in various types of brain tumor tissue. The results showed that all SLITRK genes are differentially expressed in brain tumors, including astrocytoma, oligodendroglioma, glioblastoma, medulloblastoma, and supratentorial primitive neuroectodermal tumor (PNET). Particularly interesting findings were that SLITRK3 expression was enhanced in tissue from several different types of tumors and SLITRK6 expression was highly selective. These results suggest that the human SLITRK genes are useful molecular indicators of brain tumor properties. PMID- 14557069 TI - Novel sequences encoding venom C-type lectins are conserved in phylogenetically and geographically distinct Echis and Bitis viper species. AB - Envenoming by Echis saw scaled vipers and Bitis arietans puff adders is the leading cause of death and morbidity in Africa due to snake bite. Despite their medical importance, the composition and constituent functionality of venoms from these vipers remains poorly understood. Here, we report the cloning of cDNA sequences encoding seven clusters or isoforms of the haemostasis-disruptive C type lectin (CTL) proteins from the venom glands of Echis ocellatus, E. pyramidum leakeyi, E. carinatus sochureki and B. arietans. All these CTL sequences encoded the cysteine scaffold that defines the carbohydrate-recognition domain of mammalian CTLs. All but one of the Echis and Bitis CTL sequences showed greater sequence similarity to the beta than alpha CTL subunits in venoms of related Asian and American vipers. Four of the new CTL clusters showed marked inter cluster sequence conservation across all four viper species which were significantly different from that of previously published viper CTLs. The other three Echis and Bitis CTL clusters showed varying degrees of sequence similarity to published viper venom CTLs. Because viper venom CTLs exhibit a high degree of sequence similarity and yet exert profoundly different effects on the mammalian haemostatic system, no attempt was made to assign functionality to the new Echis and Bitis CTLs on the basis of sequence alone. The extraordinary level of inter specific and inter-generic sequence conservation exhibited by the Echis and Bitis CTLs leads us to speculate that antibodies to representative molecules should neutralise the biological function of this important group of venom toxins in vipers that are distributed throughout Africa, the Middle East and the Indian subcontinent. PMID- 14557070 TI - Interaction of proteins with promoter elements of the human U2 snRNA genes in vivo. AB - The multicopy human U2 small nuclear (sn)RNA genes are transcribed by RNA polymerase (pol) II and contain two major promoter elements upstream of the transcription start site: an essential proximal sequence element (PSE) at around 55 and a distal sequence element (DSE) at around -220. We have carried out an in vivo footprinting analysis on these genes, and the results suggest that most, if not all, of the U2 gene promoters are bound by factors in interphase. Both the DSE and the PSE are protected from digestion, and the pattern of methylation protection over the DSE is virtually identical to that obtained in vitro using nuclear extract. Our results also indicate that the DNA between the PSE and the transcription start site is distorted and that proteins interact with the promoter between -20 and -33. Mutation of this sequence affects both the accuracy of initiation and polymerase specificity, underlining the importance of this region in U2 gene expression. We have also analysed the pattern of protection over the DSE and PSE of the U2 genes in mitotic cells. The degree of protection over all promoter elements is drastically reduced, suggesting that loss of DNA binding factors from the promoter plays a role in the shutdown of U2 gene transcription in mitosis. PMID- 14557071 TI - Neutral ceramidase gene: role in regulating ceramide-induced apoptosis. AB - The sphingolipid, ceramide, is a natural dietary constituent and a potent mediator of apoptosis. If left undegraded, it may induce apoptosis and cause disruption of cellular integrity. A potential mechanism to prevent ceramide induced apoptosis in various organs may involve ceramidases that facilitate the degradation of ceramide. In this study, we first isolated and characterized the murine neutral ceramidase (N-CDase) gene, mapped its chromosomal location and determined its developmental and organ-specific expression. Then we used cultured mesangial cells as our in vitro model and mouse gastrointestinal (GI) tract as the in vivo model to determine the effects of an inhibitor of N-CDase, D-erythro MAPP, to delineate whether N-CDase plays a role in preventing ceramide-induced apoptosis. Our results show that: (i) the structure of the murine neutral ceramidase gene is virtually identical to that of the human gene; (ii) it is localized on chromosome 19 at bands C2-C3 that is syntenic to human chromosome 10q24-26; (iii) N-CDase expression is developmentally regulated and it is expressed at high levels in cultured mesangial cells and in specific regions of the mouse small intestine; (iv) inhibition of N-CDase by D-erythro-MAPP leads to increased ceramide levels and consequent apoptosis in cultured mesangial cells; (v) mice treated with D-erythro-MAPP alone also caused apoptosis in the small intestine; and (vi) mice treated with D-erythro-MAPP prior to feeding C2 ceramide manifest markedly elevated levels of apoptosis in the GI tract raising the possibility that neutral ceramidase plays a detoxifying role against inadvertent stimulation of ceramide-induced apoptosis in organs that come in contact with this sphingolipid. We propose that N-CDase is an essential component of an innate detoxifying mechanism to prevent ceramide-induced apoptosis. PMID- 14557072 TI - Lupin nad9 and nad6 genes and their expression: 5' termini of the nad9 gene transcripts differentiate lupin species. AB - The mitochondrial nad9 and nad6 genes were analyzed in four lupin species: Lupinus luteus, Lupinus angustifolius, Lupinus albus and Lupinus mutabilis. The nucleotide sequence of these genes confirmed their high conservation, however, higher number of nucleotide substitution was observed in the L. albus genes. Southern hybridizations confirmed the presence of single copy number of these genes in L. luteus, L. albus and L. angustifolius. The expression of nad9 and nad6 genes was analyzed by Northern in different tissue types of analyzed lupin species. Transcription analyses of the two nad genes displayed single predominant mRNA species of about 0.6 kb in L. luteus and L. angustifolius. The L. albus transcripts were larger in size. The nad9 and nad6 transcripts were modified by RNA editing at 8 and 11 positions, in L. luteus and L. angustifolius, respectively. The gene order, rps3-rpl16-nad9, found in Arabidopsis thaliana is also conserved in L. luteus and L. angustifolius mitochondria. L. luteus and L. angustifolius showed some variability in the sequence of the nad9 promoter region. The last feature along with the differences observed in nad9 mRNA 5' termini of two lupins differentiate L. luteus and L. angustifolius species. PMID- 14557073 TI - An interplay of Sp1, GKLF and CREB-2 controls human Pre-alpha-Inhibitor gene (ITIH3) transcription. AB - Pre-alpha-Inhibitor is a plasma protease inhibitor and a heterodimeric molecule whose one polypeptide chain is encoded by the ITIH3 gene. In order to understand the expression of this protein that is regulated in health and disease, we have analyzed the 5' flanking region of ITIH3, specifically focussing on its proximal promoter. A combination of methods including wild-type (wt) or mutant promoter linked to a reporter cat gene, co-transfections of cat constructs with expression plasmids for nuclear factors and electrophoretic mobility shift assays revealed that two antagonistic sets of regulatory elements and nuclear proteins are critical for the activity of this promoter. Indeed, several overlapping Sp1/Sp3 binding sites are required for a sustained activity. However, a tripartite complex including CREB-2 and two molecules of the gut-enriched, Kruppel-like factor cooperate to bind to an upstream area whose 3' end overlaps the Sp1 binding sites. The resulting competition between this tripartite complex and Sp1 results in impaired occupancy of Sp1-binding sites by Sp1 and a consequent reduction in ITIH3 transcription. Competition between Sp1 and a Kruppel-like factor for GC-rich sites has been previously reported, but this is the first description of an elaborate tripartite cooperation of two Kruppel-like factors and CREB as a key step in such a competition. PMID- 14557074 TI - Gene cloning, purification, and stoichiometry quantification of phi29 anti receptor gp12 with potential use as special ligand for gene delivery. AB - Bacterial virus phi29 is the most efficient in vitro DNA packaging system, with which up to 90% of the added DNA can be packaged into purified recombinant procapsid in vitro. The findings that phi29 virions can be assembled with the exclusive use of cloned gene products have bred a thought that phi29 has a potential to be a gene delivery vector since it is a nonpathogenic virus. gp12 of bacterial virus phi29 has been reported to be the anti-receptor that is responsible for binding the virus particle to the host cell. We cloned the gene coding gp12, overexpressed it in Escherichia coli, and purified the gene product to study the properties and functions of gp12 in virus assembly. According to SDS PloyAcrylamide Gel Electrophoresis (SDS-PAGE) analysis and N-terminal sequencing, recombinant gp12 isolated from E. coli had a molecular mass of 80 kDa, and 24 amino acids at N-terminal were cleaved after expression. The purified recombinant gp12 was incorporated into phi29 particles and converted the gp12-lacking assembly intermediates of phi29 into infectious virions in vitro. This purified protein gp12 was able to compete with infectious phi29 virions for binding to the host cell, thus inhibiting the infection by phi29. Scanning Transmission Electron Microscopy (STEM) analysis and sedimentation studies revealed that recombinant gp12 products were assembled into biologically active dimers. Analysis of the dose-response curve showed that 12 dimeric gp12 complexes were assembled onto viral particles and that each virion contained 24 copies of gp12 molecules. The results provide a basis for future research into bacteriophage-host interaction by modifying the anti-receptor protein. The ultimate goal is to re-target the bacteriophage to new host cells for the purpose of gene delivery. PMID- 14557075 TI - Isolation and characterisation of two multidrug resistance associated protein genes from maize. AB - Two genes encoding ATP-binding cassette (ABC) transporters were isolated from the crop plant Zea mays (maize). The clones, designated ZmMRP1 and ZmMRP2, were highly homologous to members of the multidrug resistance associated protein (MRP) subfamily. Genomic Southern analysis and characterisation of bacterial artificial chromosome (BAC) clones demonstrated that both genes are present in two copies in maize, which are located in proximity to each other, suggesting the occurrence of duplication events. The full-length genomic and cDNA sequences of ZmMRP1 and 2 were obtained, permitting analysis of the intron/exon structures and protein domains. Intron positions and phasing were conserved between ZmMRP1 and 2 and their closest Arabidopsis homologues. Both clones contained two copies each of the membrane spanning domains and nucleotide-binding folds diagnostic of the ABC superfamily, and ZmMRP1 contained an additional N-terminal membrane-spanning domain (MSD0) that is typical of MRP transporters but which is lacking in the most closely related Arabidopsis and rice MRPs. In contrast, ZmMRP2 and its closest rice but not Arabidopsis homologues lacked MSD0, suggesting the repeated loss of this domain in MRP family evolution. ZmMRP1 and 2 were expressed in all tissues examined but displayed distinct expression profiles in response to herbicide safeners and pro-oxidants. ZmMRP1 was induced by aminotriazole and to a lesser extent by menadione, whereas ZmMRP2 was expressed at a lower constitutive level and did not exhibit strong induction by any of the compounds tested. The characterisation of these clones represents an important step in the experimental analysis of the MRP subfamily in a monocotyledonous crop plant. PMID- 14557076 TI - Differential gene expression in genetically matched mouse melanoma cells with different metastatic potential. AB - In vitro fusion of weakly metastatic Cloudman S91 melanoma cells with macrophages from DBA/2J mice (syngeneic with Cloudman S91 melanoma) produced hybrids with metastatic potentials ranging from low to high, with more than half showing enhanced metastasis over the parental melanoma [Clin. Exp. Metastasis 16 (1998) 299]. These hybrids, derived from the same parental fusion partners, represent a unique genetically matched model for analyzing differential gene expression regulating the metastatic phenotype. We have examined the differences in gene expression in metastatic fusion hybrid compared to its parental partners, non /poorly metastatic melanoma cells and normal macrophages. An approach by selective polymerase chain reaction (PCR) amplification and display of 3' end restriction fragments of double-stranded cDNAs was used [Methods Enzymol. 303 (1999) 272]. Gene expression analyses showed an extensive set of transcripts that were up- or down-regulated in the most metastatic hybrid, H95-1, compared to the parental macrophages or melanoma cells. Sequence analyses of more than 60 of these differentially expressed cDNAs revealed significant up- or down-regulation of a number of genes known to be associated with metastasis of melanoma and other solid tumors. Some genes are found to express exclusively either in normal macrophages or in melanoma. Thirteen fragment sequences were found with no matches with GenBank search. Comparison of these gene expression patterns should be of great value in understanding the coordinate programs regulating metastasis. Further, the increased expression of gene(s) common in macrophage and fusion hybrids may be of importance in identifying the regulatory factor(s) related to macrophage-like trait, motility, a critical step of metastatic processes, in hybrids. PMID- 14557077 TI - A single amino acid substitution in MSH5 results in DNA alkylation tolerance. AB - DNA alkylation tolerance is a major concern in cancer chemotherapy. It has been suggested that mutations in DNA mismatch repair genes may result in alkylation tolerance. This alkylation tolerant phenotype is often manifested in cells lacking an O(6)-methylguanine DNA methyltransferase (MTase) activity. However, deletion of each mismatch repair gene in the MTase mutant of a model eukaryotic yeast does not result in alkylation tolerance. We previously isolated an alkylation tolerant mutant and mapped the mutation to MSH5. Here we present evidence that a single point mutation that results in a Y823H amino acid substitution, but not deletion, of the MSH5 gene is responsible for tolerance to killing by DNA alkylating agents. We also find that other preexisting amino acid variations may also enhance alkylation tolerance in the above mutation background. Since MSH5 encodes a protein homologous to DNA mismatch recognition proteins, mismatch repair genes are frequently mutated in cancers cells and, like mismatch repair genes, MSH5 is highly conserved from yeast to human, this observation suggests novel mechanisms of chemotherapeutic drug resistance that may occur in certain human cancer patients. PMID- 14557079 TI - Sometimes a cigar [magazine] is more than just a cigar [magazine]: pro-smoking arguments in Cigar Aficionado, 1992-2000. AB - Since its first issue in 1992, few periodicals have enjoyed the rapid growth and international popularity of Cigar Aficionado. Although the magazine professes to simply celebrate "the good life and the joys of cigar smoking," we argue that it serves a more insidious function; specifically, the periodical supplies readers with 7 persuasive strategies aimed at rebuking dominant anti-smoking health assertions: (a) the cigars-are-not-cigarettes argument, (b) the life-is-dangerous argument, (c) the health-benefits argument, (d) the moderation argument, (e) the old-smokers argument, (f) the bad-science argument, and (g) the good-science argument. These pro-smoking arguments ultimately serve to relieve the cognitive dissonance associated with the consumption of a potentially deadly product and to maintain a loyal readership, free from guilt or anxiety. PMID- 14557078 TI - The mouse YAF2 gene generates two distinct transcripts and is expressed in pre and postimplantation embryos. AB - Mammalian Polycomb group (PcG) proteins are known to function during the maintenance of spatially restricted expression of Hox cluster genes and cellular proliferation. To understand the molecular basis of PcG functions, it is important to identify the components of mammalian PcG complexes. We isolated mouse YAF2 as a protein that interacts with Ring1B, a known constituent of mammalian PcG complexes. We show that the murine YAF2 locus generates two different transcripts, mYAF2-a and mYAF2-b by alternative splicing of the third exons which encode two YAF2 isoforms of 179 and conceptual 60 amino acids, respectively. At least five exons encoding mYAF2 transcripts are mapped on chromosome 15E3 region. Expression of mYAF2 mRNA was observed in both pre- and postimplantation embryos. In mid-gestation embryos, mYAF2 expression is strongly seen in the region close to the surface ectoderm. Finally, biochemical evidence and colocalization studies in tissue culture cells suggest that the product of the mYAF2 gene is involved in PcG complexes together with Ring1B and/or Ring1A. PMID- 14557080 TI - Expanding the boundaries of medical education: evidence for cross-cultural exchanges. AB - PROBLEM STATEMENT AND BACKGROUND: Cross-cultural experiences are in increasing demand by both graduate and undergraduate medical students, yet the benefits of these experiences are not clearly established. METHOD: A review of the literature was conducted to identify articles on the outcomes of cross-cultural experiences. Themes were identified and categorized into domains. RESULTS: Forty-two studies were found; 27 articles used qualitative methods, nine used quantitative methods, and six used both. Most (24) were from the nursing literature, 18 were from the medical literature. All studies reported positive outcomes along four domains: students' professional development, students' personal development, medical school benefits, and host population benefits. CONCLUSIONS: Studies reviewed were primarily case controlled or case series. Future research is needed that more clearly defines outcome measures and uses more rigorous methods. Although results suggest positive outcomes in all domains, additional research is needed before cross-cultural rotations can be supported based on evidence. PMID- 14557081 TI - Recent trends in match process outcomes for U.S. senior medical students. AB - PURPOSE: It is difficult to compare match outcomes across specialties because they may change over time and data are not reported in uniform formats. METHOD: Recent U.S. student match outcomes were analyzed for 16 specialties. RESULTS: On the basis of student match rates, three specialties were consistently competitive, eight were consistently noncompetitive, and five were in flux. Unfilled position rates increased for five specialties and remained stable or decreased for 11. CONCLUSIONS: Many specialties, both competitive and noncompetitive, recently have become increasingly desirable to match participants, whereas primary care specialties have become less desirable to both U.S. students and other match participants. PMID- 14557082 TI - The impact of postgraduate training and timing on USMLE Step 3 performance. AB - PURPOSE: This study examined the extent to which differences in clinical experience, gained in postgraduate training programs, affect performance on Step 3 of the United States Medical Licensing Examination (USMLE). METHOD: Subjects in the study were 36,805 U.S. and Canadian medical school graduates who took USMLE Step 3 for the first time between November 1999 and December 2002. Regression analyses examined the relation between length and type of postgraduate training and Step 3 score after controlling for prior performance on previous USMLE examinations. RESULTS: Results indicate that postgraduate training in programs that provide exposure to a broad range of patient problems, and continued training in such areas, improves performance on Step 3. CONCLUSIONS: Study data reaffirm the validity of the USMLE Step 3 examination, and the information found in the pattern of results across specialties may be useful to residents and program directors. PMID- 14557083 TI - Trouble in academia: ten years of litigation in medical education. AB - BACKGROUND: This study looks at a disturbing increase in medical education ("ME") litigation over the past ten years (1993-2002). METHOD: Using the LEXIS/NEXIS online legal database, this study identifies and analyzes cases in which a medical student, resident, or faculty member ("ME participants") was involved in a lawsuit with their respective institution or some other aspect of ME. RESULTS: The majority of claims lodged against institutions by ME participants concern the ME participant's termination from their respective institution and allege institutional discrimination or the failure to provide adequate due process protections. CONCLUSION: Discrimination and due process claims are direct challenges to the effectiveness of an institution's policies and procedures regarding physician review, promotion, and termination. All relevant events in a student or resident's education, positive and negative, must be fully documented. The failure to establish and faithfully implement comprehensive policies and procedures leaves institutions particularly vulnerable to this most frequent cause of litigation. PMID- 14557084 TI - Educating for advocacy: exploring the source and substance of community responsive physicians. AB - PURPOSE: Physicians are called to be community responsive. But with few existing operational definitions, how can it be taught? The nature, inclination, and impact of community-responsive physicians were studied to address this educational fissure. METHOD: Case studies on eight community-responsive physicians were conducted. Data were qualitatively analyzed using NVivo software. RESULTS: One way physicians were community responsive was through advocacy. Three themes arose: (1) having knowledge of difference, social determinants of health, and the power and privilege of physicians; (2) being influenced by role models and exposure to marginalized groups; and (3) motivation to do the right thing, give back, make a difference, and be intellectually challenged. CONCLUSIONS: Educational strategies to advance the teaching of physicians to be community responsive advocates are highlighted. PMID- 14557085 TI - Identification of physician and patient attributes that influence the likelihood of screening for intimate partner violence. AB - PURPOSE: Effective assessment of intimate partner violence (IPV) demands that everyone at risk be screened. To identify potential barriers, paper-and-pencil case scenarios identified possible practitioner and patient attributes that influence IPV screening. METHOD: First-year residents responded to one of four short written scenarios describing a divorced female patient with nonlocalized abdominal pain; variables were patient's age and abdominal bruising. Residents rated their likelihood of screening for IPV and seven other screening tasks and self-assessed their competence in performing each task. Regression analyses assessed the influence of resident and patient characteristics on screening likelihood. RESULTS: Patient bruising, younger patient age, and resident self assessed competence best predicted IPV screening. Men were less likely than women to screen for IPV. CONCLUSIONS: Although most physicians receive training on IPV in medical school, barriers to IPV screening still exist. Identifying obstacles to IPV risk-assessment is an essential prerequisite for improving educational programs that promote routine IPV screening. PMID- 14557086 TI - Domestic violence: increasing knowledge and improving skills with a four-hour workshop using standardized patients. AB - PURPOSE: Domestic violence (DV) is common, yet physicians feel unprepared to address it. Educational interventions may improve the care provided to DV victims, yet the effectiveness of interventions is often unproven. METHOD: Written questions and DV-specific standardized patient (SP) checklist items from an end-of-clerkship and fourth-year comprehensive multispecialty (the Clinical Performance Examination or CPX) examinations of medical students participating in a DV workshop using SPs was compared with nonparticipants. RESULTS: DV workshop participants' and nonparticipants' written question mean scores were 93.2% and 85.8%, respectively, p =.02. End-of-clerkship SP examination DV-specific checklist scores for participants and nonparticipants was 76.3% and 60.0%, respectively, p =.002. Workshop participants scored 44.1% on the CPX DV-specific checklist items versus 35.6% for the nonparticipants, p =.01. CONCLUSION: A DV workshop improved knowledge and skills assessed four and an average of 27 weeks later. PMID- 14557087 TI - An examination of the relationship between clinical skills examination performance and performance on USMLE Step 2. AB - PURPOSE: To examine the relationship between performance on a large-scale clinical skills examination (CSE) and a high-stakes multiple-choice examination. METHOD: Two samples were used: (1) 6,372 first-taker international medical graduates (IMGs); and (2) 858 fourth-year U.S. medical students. Ninety-seven percent of IMGs and 70% of U.S. students had completed Step 2. Correlations were calculated, scatter plots produced, and regression lines estimated. RESULTS: Correlations between CSE and Step 2 ranged from .16 to .38. The observed relationship between scores confirms that CSE score information is not redundant with MCQ score information. This result was consistent across samples. CONCLUSIONS: Results suggest that the CSE assesses proficiencies distinct from those assessed by current USMLE components and therefore provides evidence justifying its inclusion in the medical licensure process. PMID- 14557088 TI - Assessing physical examination skills of senior medical students: knowing how versus knowing when. AB - PURPOSE: Literature has documented a lack of physical examination proficiency among medical students. To investigate UCLA medical students' physical examination proficiency, this study compared their performance on a multiple station Clinical Performance Examination (CPX) focused on clinical cases with that on a thorough physical examination objective structured clinical examination (OSCE) of the same organ systems. METHOD: One hundred sixty-three beginning fourth-year students participated in the study. Four organ systems were included in both the CPX and OSCE. Pearson's correlation coefficients and paired-sample t tests were conducted to determine the correlations between and differences in the two tests. RESULTS: The physical examination scores on the CPX were significantly (p <.001) lower both by organ subscale and in total than those on the OSCE. Correlation coefficients between the two tests were negligible. CONCLUSIONS: The study suggests that fourth-year medical students may demonstrate competency in directed examination of organ systems while being unable to correctly apply those examination skills to the work-up of clinical cases. PMID- 14557089 TI - Feasibility, reliability, and validity of the mini-clinical evaluation exercise (mCEX) in a medicine core clerkship. AB - PURPOSE: To determine the feasibility, reliability and validity of the mCEX when used to evaluate medical students' clinical skills in a medicine core clerkship. METHOD: In 2002, students were required to complete nine mCEX during their medicine clerkship. Mean mCEX scores were correlated with exam scores and course grades. RESULTS: 89% of targeted mCEX were completed. The reproducibility coefficient for eight mCEX was.77. Mean mCEX scores were significantly correlated with exam scores (r =.22; p =.004), inpatient (r =.43; p <.0001), outpatient (r =.35; p <.0001), and final course grades (r =.19; p =.014). CONCLUSIONS: These data support the feasibility, reproducibility, and validity of the mCEX in evaluating medicine clerkship students' clinical skills. PMID- 14557090 TI - Intersite consistency as a measurement of programmatic evaluation in a medicine clerkship with multiple, geographically separated sites. AB - PURPOSE: This study introduces "intersite consistency" as a measurement of programmatic evaluation and demonstrates its feasibility and construct validity. METHOD: Student data in our multisite, geographically separated clerkship were collected prospectively over a ten-year period (1990-2000). We calculated mean scores for each clerkship measurement and analyzed these data on both a yearly and a 10-year cumulative basis. Analyses of variance (ANOVA) and linear regression were used for statistical analysis. RESULTS: Data for 1,632 (98%) students were included in our study. During this ten-year study period, we had 22 different on-site clerkship directors at seven clerkship sites. ANOVA and linear regression of year-to-year and cumulative data did not demonstrate an effect of site on student outcomes. CONCLUSIONS: Intersite consistency can be used as one measure of programmatic evaluation for multisite clerkships. PMID- 14557091 TI - Do peer chart audits improve residents' performance in providing preventive care? AB - PURPOSE: One recommended method to evaluate residents' competence in practice based learning and improvement is chart audit. This study determined whether residents improved in providing preventive care after a peer chart audit program was initiated. METHOD: Residents audited 1,005 charts and scored their peers on 12 clinical preventive services. The mean total chart audit scores were compared across five time blocks of the 45-month study. RESULTS: Residents' performance in providing preventive care initially improved significantly but declined in the last ten months. However, their performance remained significantly higher than at the beginning. CONCLUSIONS: By auditing their peers' charts, residents improved their own performance in providing preventive care. The diffusion of innovations theory may explain the prolonged implementation phase and problems in maintaining a chart audit program. PMID- 14557092 TI - Responses of rural family physicians and their colleague and coworker raters to a multi-source feedback process: a pilot study. AB - PURPOSE: To describe responses of family physicians, their medical colleagues, and coworker raters to a multisource feedback assessment process. METHOD: Data collection tools included multisource feedback self-assessment and medical colleague, coworker, and patient rating forms; and program evaluation physician and rater questionnaires. RESULTS: The pilot study included 142 physicians and their raters, with 113 (80%) physicians completing evaluations. Positive correlations were found between familiarity scores and medical colleague and coworker mean ratings. Peer medical colleagues were significantly more familiar with physicians than were consultants. Consultants were unable to rate items most frequently. Physicians disagreed with colleague feedback more frequently. Agreement was positively correlated with scores. CONCLUSIONS: Familiarity, ability to observe physicians appropriately to rate them, and physicians' responses to feedback are factors to consider when multisource feedback is used. PMID- 14557093 TI - Assessment of a clinical performance evaluation tool for use in a simulator-based testing environment: a pilot study. AB - PURPOSE: This study assessed a clinical performance evaluation tool for use in a simulator-based testing environment. METHOD: Twenty-three subjects were evaluated during five standardized encounters using a patient simulator (six emergency medicine students, seven house officers, ten chief resident-fellows). Performance in each 15-minute session was compared with performance on an identical number of oral objective-structured clinical examination (OSCE) sessions used as controls. Each was scored by a faculty rater using a scoring system previously validated for oral certification examinations in emergency medicine (eight skills rated 1 8; passing = 5.75). RESULTS: On both simulator exams and oral controls, chief resident-fellows earned (mean) "passing" scores [sim = 6.4 (95% CI: 6.0-6.8), oral = 6.4 (95% CI: 6.1-6.7)]; house officers earned "borderline" scores [sim = 5.6 (95% CI: 5.2-5.9), oral = 5.5 (95% CI: 5.0-5.9)]; and students earned "failing" scores [sim = 4.3 (95% CI: 3.8-4.7), oral = 4.5 (95% CI: 3.8-5.1)]. There were significant differences among mean scores for the three cohorts, for both oral and simulator test arms (p <.01). CONCLUSIONS: In this pilot, a standardized oral OSCE scoring system performed equally well in a simulator-based testing environment. PMID- 14557094 TI - Introduction of patient video clips into computer-based testing: effects on item statistics and reliability estimates. AB - PURPOSE: Using patient video clips to evaluate examinees' skills in interpreting physical examination findings is possible with computer-based testing, but the psychometric properties of video-based questions are unknown. METHOD: We developed parallel test questions incorporating video clips or text descriptions of abnormal neurologic findings and administered them to 106 fourth-year medical students finishing their Neurology Clerkship. RESULTS: Overall, video-based questions had comparable difficulty and discrimination compared to analogous text based questions. Preliminary studies indicated similar reliability with text- and video-based questions. CONCLUSIONS: The inclusion of patient video clips in computer-based testing is feasible from technical, practical, and psychometric perspectives. Further study is needed to gather validity evidence for this novel question format. PMID- 14557096 TI - Team learning in medical education: initial experiences at ten institutions. AB - PURPOSE: In the midst of curricular reforms that frequently call for reducing lectures and increasing small-group teaching, there is a crisis in faculty time for teaching. This paper describes the initial experiences of ten institutions with team learning (TL), a teaching method which fosters small-group learning in a large-class setting. METHOD: After initial pilot studies at one institution, nine additional institutions implemented TL in one or more courses. RESULTS: Within 18 months, TL has been used in 40 courses (from.5% to 100% of the time) and all ten institutions will increase its use next year. CONCLUSIONS: We surmise that this relatively rapid spread of TL into the medical curriculum is due to the sound pedagogy and efficiency of TL as well as the modest financial resources and support we have provided to partner institutions. PMID- 14557095 TI - Development of a reliable multimedia, computer-based measure of clinical skills in bedside neurology. AB - PURPOSE: This study reports the development of reliable multimedia, computer based measures of bedside neurology skills. METHOD: A consortium of neurologists and medical educators (1) identified bedside skills and (2) created a computer based test. Test-item stems were multimedia clips of standardized patients. Options appeared as text. Sixty-one junior medical students responded to these items. RESULTS: The 77-item test yielded a reliability coefficient of 0.85. Subgroup item analysis resulted in reliability coefficients of 0.73 for the 20 question "pretest" (Test A) and 0.75 for the 20-question "posttest" (Test B). CONCLUSION: Two sets of test items were developed that can be used as outcome measures in studies that assess the effectiveness of educational interventions in bedside neurology. PMID- 14557097 TI - A liberal arts education as preparation for medical school: how is it valued? How do graduates perform? AB - PURPOSE: This study examines the utility of a liberal arts education on medical students' preparation and performance. METHOD: Data included a survey of admission committee members, a preadmission survey of two cohorts of students, and academic performance and extracurricular involvement during medical school. RESULTS: Some admission committee members perceived applicants with liberal arts backgrounds to have certain advantages. These students preferred "discussing issues," and showed an initial preference for the practice of psychiatry. Despite entering with lower total grade-point average and being less involved in extracurricular activities, they were more likely to receive formal commendation and be elected to Alpha Omega Alpha. CONCLUSIONS: Although similarities outweigh differences, students with liberal arts backgrounds may benefit from an educational breadth well-suited for practicing the "art of medicine." PMID- 14557099 TI - An evaluation of local item dependencies in the Medical Council of Canada Qualifying Examination Part I. AB - PURPOSE: This study investigates (a) whether items within the Multiple-Choice Questions component of the Medical Council of Canada's Qualifying Examination Part I exhibit local dependencies and (b) potential sources of such dependencies. METHOD: The dimensionality of each of six discipline-based subtests was assessed based on exploratory nonlinear factor analyses. A standardized Fisher's z statistic was used to test residual item correlations for local item dependencies. The characteristics of pairs of items flagged as possibly locally dependent were reviewed. RESULTS: Some items in the Pediatrics and Public Medicine/Community Health subtests are locally dependent; these tend to be the more difficult items on the subtests. DISCUSSION: While these results are encouraging, the possible causes and potential impacts of any local dependencies should be investigated further. PMID- 14557098 TI - Differential weighting of errors on a test of clinical reasoning skills. AB - PROBLEM STATEMENT AND BACKGROUND: Examinees can make three types of errors on the short-menu questions in the Clinical Reasoning Skills component of the Medical Council of Canada's Qualifying Examination Part I: (1) failing to select any correct responses, (2) selecting too many responses, or (3) selecting a response that is inappropriate or harmful to the patient. This study compared the information provided by equal and differential weighting of these errors. METHOD: The item response theory nominal model was applied to fit examinees' response patterns on the 1998 test. RESULTS: Differential error weighting resulted in improved model fit and increased test information for examinees in the lower half of the achievement continuum. CONCLUSION: Differential error weighting appears promising. The pass score is near the lower end of the achievement continuum; therefore, this approach may improve the accuracy of pass-fail decisions. PMID- 14557100 TI - Analysis of the relationship between score components on a standardized patient clinical skills examination. AB - PURPOSE: This work investigated the reliability of and relationships between individual case and composite scores on a standardized patient clinical skills examination. METHOD: Four hundred ninety two fourth-year U.S. medical students received three scores [data gathering (DG), interpersonal skills (IPS), and written communication (WC)] for each of 10 standardized patient cases. mGENOVA software was used for all analyses. RESULTS: Estimated generalizability coefficients were 0.69, 0.80, and 0.70 for the DG, IPS, and WC scores, respectively. The universe-score correlation between DG and WC was high (.83); those for DG/IPS and IPS/WC were not as strong (0.51 and 0.37, respectively). Task difficulty appears to be modestly but positively related across the three scores. Correlations between the person-by-task effects for DG/IPS and DG/WC were positive yet modest. The estimated generalizability coefficient for a ten-case test using an equally weighted composite DG/WC score was 0.78. CONCLUSIONS: This work allows for interpretation of correlations between (1) proficiencies measured by multiple scores and (2) sources of error that affect those scores as well as for estimation of the reliability of composite scores. Results have important implications for test construction and test validity. PMID- 14557101 TI - Correlates of performance of the ECFMG Clinical Skills Assessment: influences of candidate characteristics on performance. AB - PURPOSE: Since 1998, over 30,000 international medical graduates (IMGs) have taken the ECFMG CSA. Although candidate background variables have been shown to predict basic and clinical science performance, little work has focused on the relationships between candidate characteristics and clinical skills performance. The purpose of this study was to investigate whether candidate characteristics were related to CSA pass/fail status. METHOD: Logistic regression analyses were conducted to explore the relationships between candidate characteristics and CSA pass/fail status for a sample of 11,690 IMGs over a two-year period. RESULTS: Results indicated that gender, English language proficiency, recent medical school graduation, and clinical science performance (USMLE Step 2) were all related to clinical skills proficiency. CONCLUSIONS: The results provide additional support for the validity of CSA pass/fail decisions. PMID- 14557102 TI - Evaluation of an automated procedure for scoring patient notes as part of a clinical skills examination. AB - PROBLEM STATEMENT AND BACKGROUND: The purpose of the present study was to examine the extent to which an automated scoring procedure that emulates expert ratings with latent semantic analysis could be used to score the written patient note component of the proposed clinical skills examination (CSE). METHOD: Human ratings for four CSE cases collected in 2002 were compared to automated holistic scores and to regression-based scores based on automated holistic and component scores. RESULTS AND CONCLUSIONS: Regression-based scores account for approximately half of the variance in the human ratings and are more highly correlated with the ratings than the scores produced from the automated algorithm. Implications of this study and suggestions for follow-up research are discussed. PMID- 14557103 TI - Can the strength of candidates be discriminated based on ability to circumvent the biasing effect of prose? Implications for evaluation and education. AB - PURPOSE: Residents have greater confidence in diagnoses when indicative features are presented in medical terminology. The current study examines the implications of this result by assessing its relationship to clinical ability. METHOD: Candidates writing the Medical Council of Canada's Qualifying Examination completed six questions in which the terminology used was manipulated. The influence of aptitude was examined by contrasting groups based on performance on the medicine section of Part I. RESULTS: The difference between the candidates was greatest in the mixed conditions in which the features consistent with one diagnosis were presented in medicalese and those consistent with a second diagnosis were presented using lay terminology; weaker candidates were more biased by language than stronger candidates. CONCLUSIONS: The results suggest that the language used in presenting case histories will influence the reliability of medical examinations. Furthermore, they suggest that weaker candidates might benefit from practice in making the translation between lay terminology and medicalese. PMID- 14557105 TI - Establishing passing standards for classroom achievement tests in medical education: a comparative study of four methods. AB - PURPOSE: The purpose of this research was to evaluate the Direct Borderline standard-setting method, designed for classroom instructor use, and to compare the characteristics of this newer method to three well-established methods. Most standard-setting methods were designed for large-scale assessments, and most research has taken place in the context of high-stakes examinations. METHOD: Four absolute standard-setting methods (Nedelsky, Direct Borderline, Hofstee, and Ebel) were studied for year 1 and 2 basic science examinations. RESULTS: The Direct Borderline method produced passing scores similar to the Nedelsky method and was reproducible. The Hofstee and Ebel methods produced the lowest passing scores. Standard errors at the passing score were the same or lower for the Direct Borderline method compared with the Nedelsky method. CONCLUSIONS: The Direct Borderline method has reasonable psychometric characteristics and may be practical for faculty to use in establishing absolute passing standards for classroom achievement tests. PMID- 14557104 TI - The privileged status of prestigious terminology: impact of "medicalese" on clinical judgments. AB - PURPOSE: Health professionals frequently use medical terminology like dyspnea or nasopharyngitis. These two studies examine how the use of medical terms affects the judgments of seriousness, prevalence, and disease; and diagnostic judgments. METHOD: In study 1, a survey containing the names of 22 diseases with either a medical or lay description was completed by 47 undergraduate psychology students and 25 medical students, who were asked to judge seriousness, prevalence, and how "disease-like" it was. In study 2, undergraduate students learned four "pseudopsychiatry" conditions, each with four associated features. Features were presented in lay or medical versions. They were then tested with 18 new cases with two medical features from one condition and two lay terms from the other. RESULTS: In study 1, the medical students rated conditions as more disease-like, more serious, and less prevalent than did the psychology students. Medical descriptions were seen as significantly less common and somewhat more serious and more disease-like. In study 2, the participants rated the condition with medical features consistently more likely than the alternative, regardless of training condition. CONCLUSIONS: The specific words used to describe a feature or condition can have an impact on judgments of likelihood of disease, and, to a lesser extent, judgments of seriousness. PMID- 14557106 TI - The effect of incorporating normative data into a criterion-referenced standard setting in medical education. AB - PURPOSE: Determining standards for assessing clinical performance is a controversial issue. Purely item-based methods such as the Angoff method often produce unrealistic judgments, even when used by experienced judges. The rather unstudied compromise methods combine absolute and relative judgments and thereby incorporate normative data into criterion-based standard-setting judgments. The purpose of this study was to compare the quality and implications of standards set by three methods used for the Objective Structured Clinical Examination (OSCE). METHOD: Ninety-six judges set standards for 36 surgical year-4 undergraduate OSCE stations. All judges had normative student performance data when judgments were made with the Angoff, Ebel, or Hofstee methods. RESULTS: The Hofstee method gave more realistic cutoff scores and standard errors and better Meskauskas and Jaeger indices than the Angoff and Ebel methods. CONCLUSIONS: Medical educators setting standards for an OSCE should consider adopting the Hofstee method. PMID- 14557107 TI - Incorporating prior beliefs about selection bias into the analysis of randomized trials with missing outcomes. AB - In randomized studies with missing outcomes, non-identifiable assumptions are required to hold for valid data analysis. As a result, statisticians have been advocating the use of sensitivity analysis to evaluate the effect of varying assumptions on study conclusions. While this approach may be useful in assessing the sensitivity of treatment comparisons to missing data assumptions, it may be dissatisfying to some researchers/decision makers because a single summary is not provided. In this paper, we present a fully Bayesian methodology that allows the investigator to draw a 'single' conclusion by formally incorporating prior beliefs about non-identifiable, yet interpretable, selection bias parameters. Our Bayesian model provides robustness to prior specification of the distributional form of the continuous outcomes. PMID- 14557108 TI - Estimating haplotype frequencies and standard errors for multiple single nucleotide polymorphisms. AB - Estimating haplotype frequencies becomes increasingly important in the mapping of complex disease genes, as millions of single nucleotide polymorphisms (SNPs) are being identified and genotyped. When genotypes at multiple SNP loci are gathered from unrelated individuals, haplotype frequencies can be accurately estimated using expectation-maximization (EM) algorithms (Excoffier and Slatkin, 1995; Hawley and Kidd, 1995; Long et al., 1995), with standard errors estimated using bootstraps. However, because the number of possible haplotypes increases exponentially with the number of SNPs, handling data with a large number of SNPs poses a computational challenge for the EM methods and for other haplotype inference methods. To solve this problem, Niu and colleagues, in their Bayesian haplotype inference paper (Niu et al., 2002), introduced a computational algorithm called progressive ligation (PL). But their Bayesian method has a limitation on the number of subjects (no more than 100 subjects in the current implementation of the method). In this paper, we propose a new method in which we use the same likelihood formulation as in Excoffier and Slatkin's EM algorithm and apply the estimating equation idea and the PL computational algorithm with some modifications. Our proposed method can handle data sets with large number of SNPs as well as large numbers of subjects. Simultaneously, our method estimates standard errors efficiently, using the sandwich-estimate from the estimating equation, rather than the bootstrap method. Additionally, our method admits missing data and produces valid estimates of parameters and their standard errors under the assumption that the missing genotypes are missing at random in the sense defined by Rubin (1976). PMID- 14557109 TI - Combining biomarkers to detect disease with application to prostate cancer. AB - In early detection of disease, combinations of biomarkers promise improved discrimination over diagnostic tests based on single markers. An example of this is in prostate cancer screening, where additional markers have been sought to improve the specificity of the conventional Prostate-Specific Antigen (PSA) test. A marker of particular interest is the percent free PSA. Studies evaluating the benefits of percent free PSA reflect the need for a methodological approach that is statistically valid and useful in the clinical setting. This article presents methods that address this need. We focus on and-or combinations of biomarker results that we call logic rules and present novel definitions for the ROC curve and the area under the curve (AUC) that are applicable to this class of combination tests. Our estimates of the ROC and AUC are amenable to statistical inference including comparisons of tests and regression analysis. The methods are applied to data on free and total PSA levels among prostate cancer cases and matched controls enrolled in the Physicians' Health Study. PMID- 14557110 TI - Control for confounding in the presence of measurement error in hierarchical models. AB - Hierarchical modeling is becoming increasingly popular in epidemiology, particularly in air pollution studies. When potential confounding exists, a multilevel model yields better power to assess the independent effects of each predictor by gathering evidence across many sub-studies. If the predictors are measured with unknown error, bias can be expected in the individual substudies, and in the combined estimates of the second-stage model. We consider two alternative methods for estimating the independent effects of two predictors in a hierarchical model. We show both analytically and via simulation that one of these gives essentially unbiased estimates even in the presence of measurement error, at the price of a moderate reduction in power. The second avoids the potential for upward bias, at the price of a smaller reduction in power. Since measurement error is endemic in epidemiology, these approaches hold considerable potential. We illustrate the two methods by applying them to two air pollution studies. In the first, we re-analyze published data to show that the estimated effect of fine particles on daily deaths, independent of coarse particles, was downwardly biased by measurement error in the original analysis. The estimated effect of coarse particles becomes more protective using the new estimates. In the second example, we use published data on the association between airborne particles and daily deaths in 10 US cities to estimate the effect of gaseous air pollutants on daily deaths. The resulting effect size estimates were very small and the confidence intervals included zero. PMID- 14557111 TI - Multivariate exploratory tools for microarray data analysis. AB - The ultimate success of microarray technology in basic and applied biological sciences depends critically on the development of statistical methods for gene expression data analysis. The most widely used tests for differential expression of genes are essentially univariate. Such tests disregard the multidimensional structure of microarray data. Multivariate methods are needed to utilize the information hidden in gene interactions and hence to provide more powerful and biologically meaningful methods for finding subsets of differentially expressed genes. The objective of this paper is to develop methods of multidimensional search for biologically significant genes, considering expression signals as mutually dependent random variables. To attain these ends, we consider the utility of a pertinent distance between random vectors and its empirical counterpart constructed from gene expression data. The distance furnishes exploratory procedures aimed at finding a target subset of differentially expressed genes. To determine the size of the target subset, we resort to successive elimination of smaller subsets resulting from each step of a random search algorithm based on maximization of the proposed distance. Different stopping rules associated with this procedure are evaluated. The usefulness of the proposed approach is illustrated with an application to the analysis of two sets of gene expression data. PMID- 14557112 TI - Generalized common spatial factor model. AB - There are often two types of correlations in multivariate spatial data: correlations between variables measured at the same locations, and correlations of each variable across the locations. We hypothesize that these two types of correlations are caused by a common spatially correlated underlying factor. Under this hypothesis, we propose a generalized common spatial factor model. The parameters are estimated using the Bayesian method and a Markov chain Monte Carlo computing technique. Our main goals are to determine which observed variables share a common underlying spatial factor and also to predict the common spatial factor. The model is applied to county-level cancer mortality data in Minnesota to find whether there exists a common spatial factor underlying the cancer mortality throughout the state. PMID- 14557113 TI - The spherical deformation model. AB - Miller et al. (1994) describe a model for representing spatial objects with no obvious landmarks. Each object is represented by a global translation and a normal deformation of a sphere. The normal deformation is defined via the orthonormal spherical-harmonic basis. In this paper we analyse the spherical deformation model in detail and describe how it may be used to summarize the shape of star-shaped three-dimensional objects with few parameters. It is of interest to make statistical inference about the three-dimensional shape parameters from continuous observations of the surface and from a single central section of the object. We use maximum-likelihood-based inference for this purpose and demonstrate the suggested methods on real data. PMID- 14557114 TI - Statistical analysis of microarray data: a Bayesian approach. AB - The potential of microarray data is enormous. It allows us to monitor the expression of thousands of genes simultaneously. A common task with microarray is to determine which genes are differentially expressed between two samples obtained under two different conditions. Recently, several statistical methods have been proposed to perform such a task when there are replicate samples under each condition. Two major problems arise with microarray data. The first one is that the number of replicates is very small (usually 2-10), leading to noisy point estimates. As a consequence, traditional statistics that are based on the means and standard deviations, e.g. t-statistic, are not suitable. The second problem is that the number of genes is usually very large (approximately 10,000), and one is faced with an extreme multiple testing problem. Most multiple testing adjustments are relatively conservative, especially when the number of replicates is small. In this paper we present an empirical Bayes analysis that handles both problems very well. Using different parametrizations, we develop four statistics that can be used to test hypotheses about the means and/or variances of the gene expression levels in both one- and two-sample problems. The methods are illustrated using experimental data with prior knowledge. In addition, we present the result of a simulation comparing our methods to well-known statistics and multiple testing adjustments. PMID- 14557115 TI - Estimation of effective reproduction numbers for infectious diseases using serological survey data. AB - The effective reproduction number of an infection, denoted Re, may be used to monitor the impact of a vaccination programme. If Re is maintained below 1, then sustained endemic transmission of the infection cannot occur. In this paper we discuss methods for estimating Re from serological survey data, allowing for age and individual heterogeneity. We describe semi-parametric and parametric models, and obtain an upper bound on Re when vaccine coverage and efficacy are not known. The methods are illustrated using data on mumps and rubella in England and Wales. PMID- 14557116 TI - Attenuation caused by infrequently updated covariates in survival analysis. AB - This paper deals with hazard regression models for survival data with time dependent covariates consisting of updated quantitative measurements. The main emphasis is on the Cox proportional hazards model but also additive hazard models are discussed. Attenuation of regression coefficients caused by infrequent updating of covariates is evaluated using simulated data mimicking our main example, the CSL1 liver cirrhosis trial. We conclude that the degree of attenuation depends on the type of stochastic process describing the time dependent covariate and that attenuation may be substantial for an Ornstein Uhlenbeck process. Also trends in the covariate combined with non-synchronous updating may cause attenuation. Simple methods to adjust for infrequent updating of covariates are proposed and compared to existing techniques using both simulations and the CSL1 data. The comparison shows that while existing, more complicated methods may work well with frequent updating of covariates the simpler techniques may have advantages in larger data sets with infrequent updatings. PMID- 14557117 TI - The Cleveland Clinic Foundation. PMID- 14557118 TI - Surgery in the aged. PMID- 14557119 TI - Molecular biology of aging. AB - Aging seems to reflect a patterned program of decline in functional reserves and a reduced ability to maintain homeostasis. Advances in molecular biology have provided new insights into the process of aging at the cellular level. In this review, a few selected models of aging that illustrate the molecular methods and techniques used in aging research are discussed. PMID- 14557120 TI - Fluid and electrolytes in the aged. AB - OBJECTIVE: To review the physiological changes in fluid and electrolytes that occur in aging. DATA SOURCES: Data collected for this review were identified from a MEDLINE database search of the English-language literature. The indexing terms were fluids, intravenous fluids, fluid resuscitation, fluid management, perioperative, electrolytes, aged, elderly, hemodynamics, hyponatremia, hypernatremia, hypocalcemia, hypercalcemia, hypomagnesemia, hypermagnesemia, hypophosphatemia, hypokalemia, and hyperkalemia. Relevant references from articles obtained by means of the above search terms were also used. STUDY SELECTION: All pertinent studies were included. Only articles that were case presentations or did not specifically address the topic were excluded. DATA SYNTHESIS: The fastest-growing segment of the population in the United States is individuals 65 years or older. It is imperative that health care professionals review the physiological changes that manifest during the aging process. Fluids and electrolytes are important perioperative factors that undergo age-related changes. These changes include impaired thirst perception; decreased glomerular filtration rate; alterations in hormone levels, including antidiuretic hormone, atrial natriuretic peptide, and aldosterone; decreased urinary concentrating ability; and limitations in excretion of water, sodium, potassium, and acid. CONCLUSIONS: There are age-related alterations in the homeostatic mechanisms used to maintain electrolyte and water balance. Health care providers must familiarize themselves with these alterations to guide treatment of this growing population. PMID- 14557121 TI - Surgery in the aged population: surgical oncology. PMID- 14557122 TI - Physiological features of aging persons. PMID- 14557124 TI - Laparoscopic surgery: an excellent approach in elderly patients. AB - HYPOTHESIS: A review of the literature will show that laparoscopy is safe and effective for the treatment of surgical diseases in elderly patients. DATA SOURCES: An electronic search using the PubMed and MEDLINE databases was performed using the term laparoscopy in elderly patients. Literature published in the English language in the past decade was reviewed. Pertinent references from articles and books not identified by the search engines were also retrieved. Relevant surgical textbooks were also reviewed. STUDY SELECTION: All relevant studies that could be obtained regardless of the study design were included. DATA EXTRACTION: All studies that contained material applicable to the topic were considered. Data on patient characteristics and surgical outcomes were abstracted. DATA SYNTHESIS: Sixteen studies evaluated laparoscopic cholecystectomy in the elderly. Compared with open cholecystectomy, elderly patients undergoing the laparoscopic procedure had a lower incidence of complications and a shorter hospitalization. In the 4 studies reporting the results of laparoscopic antireflux surgery in the elderly, the morbidity, mortality, and length of hospital stay were similar to those of younger patients. The elderly had equally good postoperative symptom relief. Ten reports of laparoscopic colon resection in the elderly demonstrated earlier return of bowel function, shorter hospitalization, and less cardiopulmonary morbidity. CONCLUSIONS: Despite underlying comorbidities, individuals older than 65 years tolerate laparoscopic procedures extremely well. Complications and hospitalization are lower than in open procedures. Surgeons need to inform primary care physicians of the excellent result of laparoscopic procedures in the elderly to encourage earlier referrals. PMID- 14557123 TI - Outcomes of ostomy procedures in patients aged 70 years and older. AB - HYPOTHESIS: Patients aged 70 years and older undergo proportionately more emergency and permanent fecal ostomy procedures than younger patients. Older patients have comparable short-term outcomes in morbidity and mortality, with adverse outcomes dependent on comorbid conditions and timing of the procedure rather than age alone. Older patients should be treated similarly to younger patients in terms of subsequent ostomy takedown, if an acceptable operative risk. DESIGN: A retrospective review of our facility's experience with fecal ostomies between 1992 and 2002 was performed to determine the effect of advanced age on surgical outcome measures. SETTING: A tertiary managed care medical center. PATIENTS: Three hundred eighty-three consecutive patients who underwent new fecal ostomy procedures between October 1, 1992, and October 1, 2002. One hundred three patients were aged 70 years or older (mean age, 76.4 years), and 280 patients were younger than 70 years (mean age, 49.6 years). There were 220 elective procedures and 163 emergency procedures. Outcome was analyzed between the 2 age groups. MAIN OUTCOME MEASURES: Indications for ostomy, type of ostomy, preoperative comorbidity, postoperative morbidity and mortality, length of intensive care unit and hospital stay, and subsequent ostomy takedown success. RESULTS: Three hundred eighty-three new fecal ostomies were created. The diagnosis leading to creation of the ostomy was more often malignancy in older patients (74.8%) compared with younger patients (45.0%). Both age groups underwent a similar proportion of emergency procedures (older vs younger patients, 43.7% vs 42.1%; P=.07), but more older patients were left with permanent stomas (59.2% vs 41.1%, P=.002). Older patients also had more preoperative comorbidities (P=.001), higher American Society of Anesthesiologists scores (P=.001), longer hospital stays (P=.04), and more postoperative complications. Thirty-day mortality was 6.8% in the older group vs 0.4% in the younger group (P=.001). Fewer older patients were eligible for ostomy reversal (41.1% vs 59.2%), and a smaller proportion of eligible older patients actually underwent the reversal procedure (78.7% vs 95.2%). The complication rate associated with ostomy reversal was not significantly different in the 2 age groups (P=.002). CONCLUSIONS: Patients aged 70 and older undergo proportionately more permanent fecal ostomy procedures than younger patients, with longer hospital stays, more postoperative complications, and higher mortality rates. However, surgical outcome measures in older patients following ostomy procedures remain within acceptable standards. Furthermore, older patients tolerate ostomy reversal with minimal morbidity and should not be denied consideration based on age alone if an eligible candidate. PMID- 14557125 TI - Transplantation in elderly patients. PMID- 14557126 TI - Trauma in the elderly patient. PMID- 14557127 TI - Aesthetic and reconstructive surgery in the aging patient. PMID- 14557128 TI - Inpatient surgery in California: 1990-2000. AB - BACKGROUND: The practice environment for surgery is changing. However, little is known regarding the trends or current status of inpatient surgery at a population level. HYPOTHESIS: Inpatient surgical care has changed significantly over the last 10 years. DESIGN: Longitudinal analysis of California inpatient discharge data (January 1, 1990, through December 31, 2000). SETTING: All 503 nonfederal acute care hospitals in California. PATIENTS: All inpatients undergoing general, vascular, and cardiothoracic surgery in California from January 1, 1990, through December 31, 2000, were obtained. MAIN OUTCOME MEASURES: Volume, mean age, comorbidity profile, length of hospital stay, and in-hospital mortality were obtained for inpatient general, vascular, and cardiothoracic surgical procedures performed during the period 1990 to 2000. Rates of change and trends were evaluated for the 10-year period. RESULTS: Between January 1, 1990, and December 31, 2000, 1.64 million surgical procedures were performed. The number of surgical procedures increased 20.4%, from 135,795 in 1990 to 163,468 in 2000. Overall, patients were older and had more comorbid disease in 2000 compared with 1990. Both crude and adjusted (by type of operation) in-hospital mortality decreased from 3.9% in 1990 to 2.75% (P<.001) and 2.58% (P<.001), respectively, in 2000. Length of hospital stay decreased over the period for all operations analyzed. CONCLUSIONS: The total number of inpatient general, vascular, and cardiothoracic surgical procedures has increased over the past decade. Furthermore, our findings indicate that the outcomes of care (eg, in-hospital mortality and length of hospital stay) for patients who undergo general, vascular, and cardiothoracic surgical procedures have improved. However, continued evaluations at the population level are needed. PMID- 14557129 TI - Pretransplantation soluble adhesion molecule expression predicts outcome after living donor renal transplantation. AB - HYPOTHESIS: Occult pretransplantation systemic inflammation will identify patients at risk for poor outcomes after renal transplantation. DESIGN: Retrospective cohort study. Adhesion molecule levels were measured in pretransplantation serum samples from 86 recipients. Univariate and multivariate analyses were conducted to assess a possible correlation between serum adhesion molecule level and outcome. SETTING: University referral center. MAIN OUTCOME MEASURES: Allograft rejection and survival. RESULTS: Patients with low levels of vascular cell adhesion molecule 1 had less graft rejection (P=.007). Low levels of vascular cell adhesion molecule 1 independently predicted decreased rejection (relative risk, 0.17; P=.01), and high levels of vascular cell adhesion molecule 1 independently predicted graft loss (relative risk, 3.83; P=.02). Similar correlations were observed for intercellular adhesion molecule 1. CONCLUSIONS: Decreased pretransplantation adhesion molecule expression correlates with less rejection, and increased levels correlate with graft loss. Assessment of pretransplantation inflammatory status may be useful in optimizing immunosuppression therapy. PMID- 14557130 TI - Excellent short-term results with steroid-free maintenance immunosuppression in low-risk simultaneous pancreas-kidney transplantation. AB - HYPOTHESIS: Steroid avoidance is possible in simultaneous pancreas-kidney transplantation with the use of newer immunosuppressive agents and induction therapy. DESIGN: A retrospective consecutive case review. SETTING: A university tertiary referral center. PATIENTS: Medical records of 40 consecutive patients who underwent pancreas-kidney transplantation from November 2000 to July 2002 were reviewed. INTERVENTION: The immunosuppression protocol used in this series of patients consisted of Thymoglobulin induction combined with mycophenolate mofetil, tacrolimus, and sirolimus for maintenance immunosuppression. Steroids were used as pretreatment only, given with Thymoglobulin, and were typically discontinued by postoperative week 1. MAIN OUTCOME MEASURES: Graft and patient survival rates, rejection rates of the kidney or pancreas, infection rates, and surgical complication rates. RESULTS: Patient, kidney, and pancreas survival rates were 95.0%, 92.5%, and 87.5%, respectively. Biopsy-proven pancreas rejection rates at 1 and 3 months' posttransplantation were 2.5%. Kidney rejection rates at 1 and 3 months were 2.5%. Steroids were given only to patients with documented transplant rejection. Surgical and medical complications were no different from earlier protocols. CONCLUSIONS: Immunosuppression protocols that do not include maintenance steroids have shown minimal rejection in the first 3 months and equivalent patient and graft survival rates compared with protocols that use steroids. The potential beneficial long-term impact of steroid avoidance will require further study. PMID- 14557131 TI - Cervical spinal cord injury and the need for cardiovascular intervention. AB - HYPOTHESIS: The level of cervical spinal cord injury (CSCI) can be used to predict the need for a cardiovascular intervention. DESIGN: Retrospective review. Data included level of spinal cord injury, Injury Severity Score, lowest heart rate, and systolic blood pressure in the first 24 hours and intensive care unit course. The level of CSCI was divided into high (cord level C1-C5) or low (cord level C6-C7). Neurogenic shock was defined as bradycardia with hypotension. Statistical analysis was performed with the t test and the chi2 test. SETTING: Level I trauma center. PATIENTS: The patients studied were those with quadriplegia who experienced a CSCI and were admitted to the hospital between December 1, 1993, and October 31, 2001. INTERVENTIONS: Pressors, chronotropic agents, and pacemakers.Main Outcome Measure Use of a cardiovascular intervention in the presence of neurogenic shock. RESULTS: Eighty-three patients met the criteria for CSCI and quadriplegia, 62 in the high (C1-C5) and 21 in the low (C6 C7) level. There was no significant difference between the 2 groups in mean +/- SD age (38.2+/-17.8 vs 34.7+/-15.6 years; P=.43), mean +/- SD Injury Severity Score (35.7+/-17.5 vs 32.5+/-11.2; P=.44), mean +/- SD admission base deficit ( 0.7+/-3.6 vs 0.7+/-2.7; P=.06), or mortality (12 [19%] of 62 patients vs 2 [10%] of 21 patients; P=.29). Neurogenic shock was present in 19 (31%) of the 62 patients with high CSCI and in 5 (24%) of the 21 patients with low CSCI (P=.56). There was a marked difference in the use of a cardiovascular intervention between those with a high and those with a low CSCI: 15 (24%) of 62 patients vs 1 (5%) of 21 patients (P=.02). Two patients with C1 through C5 spinal cord injuries required cardiac pacemakers. CONCLUSIONS: There was no significant difference in the frequency of neurogenic shock by injury level. Patients with a high CSCI (C1 C5) had a significantly greater requirement for a cardiovascular intervention compared with patients with lower injuries (C6-C7). PMID- 14557132 TI - Use of the radial artery for hemodialysis access. AB - HYPOTHESIS: Under limited conditions, use of the radial artery (RA) for hemodialysis access in patients with end-stage renal disease is safe and effective. METHODS: The nondominant upper extremities of 211 consecutive patients with end-stage renal disease were evaluated with duplex ultrasonography to assess RA and ulnar artery (UA) flow contributions to their hands. Diameters and peak systolic velocities were measured in each artery, from which peak flows were calculated. Arteries of less than 2.0 mm in diameter were deemed unusable for access procedures or inadequate as the sole supply to the hand. Flow rates within 20% of each other were considered equivalent, and the RA was dominant if its flow exceeded that of the UA by 20%. Radial arteries with peak flow rates of less than 125 mL/min were judged inadequate, and RAs were not used in patients with peak UA flow rates of less than 100 mL/min. RESULTS: The RA flow was equivalent to the UA flow in 56 patients and was dominant in 120. Flow through the RA was adequate in 166 patients, but 41 had insufficient UA flow contributions and 15 had unsatisfactory UA diameters. Overall, 25 patients had inadequate RA diameters, 72 patients had inadequate UA diameters, and 16 patients had insufficient calibers of both vessels. CONCLUSIONS: The RA carried sufficient flow to support a shunt or fistula in 78.7% of patients, but it would have been unwise to use it in about one third of those cases because of potentially insufficient perfusion to the hand by the UA. The RA could therefore be safely used in 49.8% of patients with end-stage renal disease evaluated for hemodialysis access. PMID- 14557133 TI - Melanoma in the elderly patient. AB - BACKGROUND: The incidence of cutaneous melanoma is rising steadily, and the rate of increase is among the highest for any form of cancer. Although the reliability of age as a prognostic factor is debatable, several studies suggest that age has an important prognostic use. HYPOTHESIS: Age alone does not predict a poor prognosis in the older patient with melanoma. SETTING: University teaching hospital. METHODS: A retrospective review was undertaken to identify patients aged 65 years or older with intermediate-thickness melanoma (1-4 mm). Two hundred thirteen such patients were identified. Data are given as mean +/- SD. RESULTS: The mean age was 72.2+/-6.1 years. The mean follow-up was 49 months. By univariate analysis, the mean disease-free survival (DFS) and overall survival (OS) for lymph node-positive patients was 36.0+/-9.6 and 56.0+/-10.6 months, respectively. The mean DFS for node-negative patients was 155.0+/-9.8 months, and the mean OS was 166.0+/-9.2 months (P<.001 for both). The mean DFS and OS for women were 151.0+/-11.2 and 163.0+/-10.9 months, respectively. In contrast, men had 116.0+/-9.5 months' DFS and 127.0+/-9.0 months' OS (P=.01 for both). By multivariate analysis, lymph node status was the most predictive variable for DFS and OS (P<.001 for both). Sex tended to affect OS (P=.02) but did not achieve prognostic significance on DFS (P=.09). Other factors such as location, ulceration, histological type, and mitoses per square millimeter failed to show any prognostic significance. Stratification into 3 age groups (65-70, 71-80, and >80 years) had no significant effect on DFS (P=.95) or OS (P=.92). CONCLUSIONS: Lymph node status is the most important prognostic factor in older patients with intermediate-thickness melanoma. Identification of high-risk factors may help stratify these patients for recommendation of more aggressive treatment or adjuvant therapies. Among these patients, age alone was not a significant prognostic factor in the clinical management of melanoma. PMID- 14557134 TI - Incision length for standard thyroidectomy and parathyroidectomy: when is it minimally invasive? AB - HYPOTHESIS: Current techniques for open conventional thyroidectomy or parathyroidectomy have evolved to enable a shorter incision (main proposition), and the length of the incision is influenced by objective factors. DESIGN: Case series. SETTING: University referral center. PATIENTS AND INTERVENTION: Retrospective study of the most recent 200 primary consecutive routine thyroid and parathyroid operations (excluding neck dissections). MAIN OUTCOME MEASURES: The length of incision was routinely measured with a ruler before the incision. Univariate and multivariate analysis was performed to distinguish variables affecting length of incision. RESULTS: Mean length of the incision was 5.5 cm for total thyroidectomy, 4.6 cm for lobectomy, and 3.5 cm for parathyroidectomy (P<.001). It was 4.1 cm for bilateral parathyroid exploration, but was reduced to 3.2 and 2.8 cm for unilateral (P<.001) and focal (P<.001) explorations, respectively. By multiple regression analysis, thyroid specimen volume and patient body mass index were independent predictors of incision length in thyroidectomy. Extent of exploration and resident training level were independent predictors of incision length in parathyroidectomy. CONCLUSIONS: Current techniques for open conventional thyroidectomy or parathyroidectomy have evolved to enable a shorter incision. Thyroid volume, patient body mass index, extent of the planned parathyroid exploration, and the resident clinical training stage are important variables for incision length in open operation and should be taken into account when minimally invasive thyroidectomy and parathyroidectomy are evaluated. PMID- 14557135 TI - RNA silencing in surgical disease. PMID- 14557136 TI - Surgery in Jamaica. PMID- 14557137 TI - Surgical mentors: from bosom buddies to Baghdad by the bay. PMID- 14557138 TI - Image of the month. Giant serous microcystic cystadenoma of the pancreas. PMID- 14557139 TI - Patients with breast cancer and positive sentinel nodes: axillary dissection or axillary radiotherapy? PMID- 14557140 TI - La vie. PMID- 14557141 TI - Mechanism of new antipsychotic medications: occupancy is not just antagonism. AB - Antagonism of D2-like dopamine receptors is the putative mechanism underlying the antipsychotic efficacy of psychotropic drugs. Positron emission tomographic studies suggest that the antipsychotic effect of dopamine receptor antagonists occurs within a therapeutic window between 60% and 80% (striatal) D2 receptor occupancy. The incidence of extrapyramidal side effects increases above the 80% threshold. However, the novel atypical antipsychotic drug, aripiprazole, occupies up to 95% of striatal D2-like dopamine receptors at clinical doses, and the incidence of extrapyramidal side effects with aripiprazole is no higher than with placebo. The most likely explanation for this finding is aripiprazole's weak partial agonism at D2-like dopamine receptors. This particular pharmacologic feature characterizes a new class of atypical antipsychotics that does not match the original concept of a therapeutic occupancy window for antagonist antipsychotics. When not involving pure antagonists, it implies a need to adjust the expected receptor occupancy (measured using positron emission tomography) for the therapeutic window. PMID- 14557142 TI - Relationship between antidepressant medication treatment and suicide in adolescents. AB - CONTEXT: A decade of increasing antidepressant medication treatment for adolescents and corresponding declines in suicide rates raise the possibility that antidepressants have helped prevent youth suicide. OBJECTIVE: To evaluate the relationship between regional changes in antidepressant medication treatment and suicide in adolescents from 1990 to 2000. DESIGN: Analysis of prescription data from the nation's largest pharmacy benefit management organization, national suicide mortality files, regional sociodemographic data from the 1990 and 2000 US Census, and regional data on physicians per capita. PARTICIPANTS: Youth aged 10 to 19 years who filled a prescription for antidepressant medication and same-aged completed suicides from 588 three-digit ZIP code regions in the United States. MAIN OUTCOME MEASURES: The relationship between regional change in antidepressant medication treatment and suicide rate stratified by sex, age group, regional median income, and regional racial composition. RESULTS: There was a significant adjusted negative relationship between regional change in antidepressant medication treatment and suicide during the study period. A 1% increase in adolescent use of antidepressants was associated with a decrease of 0.23 suicide per 100 000 adolescents per year (beta = -.023, t = -5.14, P<.001). In stratified adjusted analyses, significant inverse relationships were present among males (beta = -.032, t = -3.81, P<.001), youth aged 15 to 19 years (beta = -.029, t = 3.43, P<.001), and regions with lower family median incomes (beta = -.023, t = 3.73, P<.001). CONCLUSIONS: An inverse relationship between regional change in use of antidepressants and suicide raises the possibility of a role for using antidepressant treatment in youth suicide prevention efforts, especially for males, older adolescents, and adolescents who reside in lower-income regions. PMID- 14557143 TI - Magnetic resonance imaging of mediodorsal, pulvinar, and centromedian nuclei of the thalamus in patients with schizophrenia. AB - BACKGROUND: Postmortem and magnetic resonance imaging (MRI) data have suggested volume reductions in the mediodorsal (MDN) and pulvinar nuclei (PUL) of the thalamus. The centromedian nucleus (CMN), important in attention and arousal, has not been previously studied with MRI. METHODS: A sample of 41 patients with schizophrenia (32 men and 9 women) and 60 healthy volunteers (45 men and 15 women) underwent assessment with high-resolution 1.2-mm thick anatomical MRI. Images were differentiated to enhance the edges and outline of the whole thalamus, and the MDN, PUL, and CMN were outlined on all slices by a tracer masked to diagnostic status. RESULTS: Significantly smaller volumes of the MDN and PUL were found in patients with schizophrenia compared with controls. Volume relative to brain size was reduced in all 3 nuclei; differences in relative reduction did not differ among the nuclei. The remainder of the thalamic volume (whole thalamus minus the volume of the 3 delineated nuclei) was not different between schizophrenic patients and controls, indicating that the volume reduction was specific to these nuclei. Volume relative to brain size was reduced in all 3 nuclei and remained significant when only patients who had never been exposed to neuroleptic medication (n = 15) were considered. For the MDN, women had larger relative volumes than men among controls, but men had larger volumes than women among schizophrenic patients. CONCLUSIONS: Three association regions of the thalamus that have reciprocal connectivity to schizophrenia-associated regions of the cortex have significantly smaller volumes on MRI in patients with schizophrenia. PMID- 14557144 TI - Longitudinal trajectories of depression and anxiety in a prospective community study: the Zurich Cohort Study. AB - BACKGROUND: The dearth of long-term follow-up studies of community-based samples and differences in methodology in existing studies highlight the need for research designed to examine the stability, comorbidity, and diagnostic thresholds of depression and anxiety in the community. METHODS: Prospective study of a community-based cohort aged 19 and 20 years from the canton of Zurich, Switzerland. Semistructured diagnostic interviews were administered by clinically experienced interviewers at 5 assessment points during a 15-year period. The format of the interview permitted assessment of major mental disorders at both the diagnostic and subthreshold levels. RESULTS: Comorbid anxiety and depression tended to be far more persistent than either syndrome alone. Individuals with anxiety states alone tended to develop either depression alone or comorbid anxiety and depression as they progressed through adulthood. In contrast, depression alone and depression comorbid with anxiety tended to be more stable than anxiety alone over time. The patterns of stability were similar for subthreshold- and threshold-level disorders. CONCLUSIONS: These findings have important implications for classification and treatment of affective disorders. The greater stability of comorbid anxiety and depression than either disorder alone illustrates the importance of further investigation of comorbid states compared with noncomorbid states in etiologic and treatment research. The persistence of subthreshold-level depression and anxiety from early to mid adulthood also suggests the importance of characterizing the continuum of expression of depression and anxiety rather than adhering to strict diagnostic thresholds. PMID- 14557145 TI - Transcranial magnetic stimulation in the treatment of depression: a double-blind, placebo-controlled trial. AB - BACKGROUND: High-frequency left-sided repetitive transcranial magnetic stimulation (HFL-TMS) has been shown to have antidepressant effects in double blind trials. Low-frequency stimulation to the right prefrontal cortex (LFR-TMS) has also shown promise, although it has not been assessed in treatment-resistant depression and its effects have not been compared with those of HFL-TMS. OBJECTIVE: To prospectively evaluate the efficacy of HFL-TMS and LFR-TMS in treatment-resistant depression and compared with a sham-treated control group. DESIGN: A double-blind, randomized, sham-controlled trial. SETTING: Two general psychiatric services. PARTICIPANTS: Sixty patients with treatment-resistant depression who had failed to respond to therapy with multiple antidepressant medications were divided into 3 groups of 20 that did not differ in age, sex, or any clinical variables. All patients completed the double-blind phase of the study. INTERVENTIONS: Twenty 5-second HFL-TMS trains at 10 Hz and five 60-second LFR-TMS trains at 1 Hz were applied daily. Sham stimulation was applied with the coil angled at 45 degrees from the scalp, resting on the side of one wing of the coil. Main Outcome Measure Score on the Montgomery-Asberg Depression Rating Scale. RESULTS: There was a significant difference in response among the 3 groups (F56,2 = 6.2), with a significant difference between the HFL-TMS and sham groups and between the LFR-TMS and sham groups (P<.005 for all) but not between the 2 treatment groups. Baseline psychomotor agitation predicted successful response to treatment. CONCLUSIONS: Both HFL-TMS and LFR-TMS have treatment efficacy in patients with medication-resistant major depression. Treatment for at least 4 weeks is necessary for clinically meaningful benefits to be achieved. Treatment with LFR-TMS may prove to be an appropriate initial repetitive TMS strategy in depression taking into account safety, tolerability, and efficacy considerations. PMID- 14557146 TI - Mild depressive symptoms are associated with amplified and prolonged inflammatory responses after influenza virus vaccination in older adults. AB - BACKGROUND: Depression is associated with enhanced production of proinflammatory cytokines that influence a spectrum of conditions associated with aging, including cardiovascular disease, osteoporosis, arthritis, type 2 diabetes mellitus, certain cancers, periodontal disease, frailty, and functional decline. In this prospective community study, we assessed the relationship between depressive symptoms and changes in inflammatory response after an influenza virus vaccination. METHODS: To study the dynamics of interleukin (IL) 6 levels in plasma in response to an immunological challenge, we obtained blood samples in 119 older adults (mean age, 71.21 +/- 8.68 years [SD]) immediately before an annual influenza vaccination and again 2 weeks later. The short form of the Beck Depression Inventory, completed at these same times, provided information on depressive symptoms. RESULTS: The number of depressive symptoms in this sample was low on average before vaccination (mean +/- SD number of symptoms, 3.07 +/- 3.09) and did not change significantly after vaccination. Participants with more depressive symptoms had higher levels of IL-6 before and after vaccination than did those who reported fewer symptoms; moreover, individuals reporting more depressive symptoms also showed an increase in plasma IL-6 levels 2 weeks later, while there was little change in IL-6 levels among those reporting few or no symptoms. CONCLUSIONS: Even a modest number of depressive symptoms may sensitize the inflammatory response system in older adults and produce amplified and prolonged inflammatory responses after infection and other immunological challenges. Sustained and/or amplified inflammatory responses could accelerate a range of age-related diseases. PMID- 14557147 TI - A polymorphism in the serotonin receptor 3A (HTR3A) gene and its association with harm avoidance in women. AB - BACKGROUND: The brain neurotransmitter serotonin is known to affect various aspects of human behavior, including personality traits. Serotonin receptor type 3 is a ligand-gated channel encoded by 2 different subunit genes, HTR3A and HTR3B. A polymorphism (C178T) in the 5' region of the HTR3A gene has recently been identified and suggested to be of functional importance. OBJECTIVE: To elucidate the possible association between the C178T polymorphism in the HTR3A gene and personality traits in women. DESIGN: Two independent samples of 35- to 45-year-old Swedish women were recruited using the population register. Sample 1 (n = 195) was assessed via the Karolinska Scales of Personality and the Temperament and Character Inventory; sample 2 (n = 175) was assessed using the latter only. Both samples were genotyped with respect to the C178T polymorphism in the HTR3A gene. The A1596G polymorphism in the same gene was also investigated. RESULTS: A significant association between C178T genotype and the Temperament and Character Inventory factor harm avoidance was observed in sample 1 (corrected for multiple comparisons P =.04); this finding was subsequently replicated in sample 2 (P =.004) (pooled populations: P<.001). In the pooled sample, all harm avoidance subscales were found to be significantly associated with the C178T polymorphism: anticipatory worry (P =.001), fear of uncertainty (P<.001), shyness (P<.001), and fatigability and asthenia (P =.008). In addition, a significant association was found in sample 1 between the C178T polymorphism and the Karolinska Scales of Personality nonconformity factor (corrected P =.002), including the subscales of social desirability (P<.001), indirect aggression (P =.002), verbal aggression (P =.05), and irritability (P<.001). Participants homozygous for the less common T allele (<4%) differed from the remaining women by displaying lower ratings on harm avoidance and nonconformity. CONCLUSION: The C178T polymorphism in the HTR3A gene may affect the personality trait of harm avoidance in women. PMID- 14557148 TI - A randomized controlled trial of cognitive therapy, a self-help booklet, and repeated assessments as early interventions for posttraumatic stress disorder. AB - BACKGROUND: It is unclear what psychological help should be offered in the aftermath of traumatic events. Similarly, there is a lack of clarity about the best way of identifying people who are unlikely to recover from early posttraumatic symptoms without intervention. OBJECTIVE: To determine whether cognitive therapy or a self-help booklet given in the initial months after a traumatic event is more effective in preventing chronic posttraumatic stress disorder (PTSD) than repeated assessments. DESIGN: Randomized controlled trial. Patients Motor vehicle accident survivors (n = 97) who had PTSD in the initial months after the accident and met symptom criteria that had predicted persistent PTSD in a large naturalistic prospective study of a comparable population. SETTING: Patients were recruited from attendees at local accident and emergency departments. INTERVENTIONS: Patients completed a 3-week self-monitoring phase. Those who did not recover with self-monitoring (n = 85) were randomly assigned to receive cognitive therapy (n = 28), a self-help booklet based on principles of cognitive behavioral therapy (n = 28), or repeated assessments (n = 29). MAIN OUTCOME MEASURES: Symptoms of PTSD as assessed by self-report and independent assessors unaware of the patient's allocation. Main assessments were at 3 months (posttreatment, n = 80) and 9 months (follow-up, n = 79). RESULTS: Twelve percent (n = 12) of patients recovered with self-monitoring. Cognitive therapy was more effective in reducing symptoms of PTSD, depression, anxiety, and disability than the self-help booklet or repeated assessments. At follow-up, fewer cognitive therapy patients (3 [11%]) had PTSD compared with those receiving the self-help booklet (17 [61%]; odds ratio, 12.9; 95% confidence interval, 3.1-53.1) or repeated assessments (16 [55%]; odds ratio, 10.3; 95% confidence interval, 2.5 41.7). There was no indication that the self-help booklet was superior to repeated assessments. On 2 measures, high end-state functioning at follow-up and request for treatment, the outcome for the self-help group was worse than for the repeated assessments group. CONCLUSIONS: Cognitive therapy is an effective intervention for recent-onset PTSD. A self-help booklet was not effective. The combination of an elevated initial symptom score and failure to improve with self monitoring was effective in identifying a group of patients with early PTSD symptoms who were unlikely to recover without intervention. PMID- 14557149 TI - Early reactions to cannabis predict later dependence. AB - CONTEXT: While there is a growing literature on the linkages between early subjective responses to nicotine and alcohol and later risks of nicotine or alcohol dependence, to date there has been no study of this issue in relation to cannabis. OBJECTIVE: To examine the extent to which subjective responses to early (prior to the age of 16 years) cannabis use were associated with subsequent cannabis dependence in a birth cohort studied to the age of 21 years. DESIGN: Data on early (prior to the age of 16 years) subjective reactions to cannabis use and subsequent cannabis dependence were gathered over the course of the Christchurch Health and Development Study, a 21-year longitudinal study of a birth cohort of children born in Christchurch, New Zealand. SETTING: General community sample. PARTICIPANTS: Members of a population-based birth cohort (86.5% white, 11.3% New Zealand Maori, and 2.2% Pacific Island). Main Outcome Measure Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition cannabis dependence (for those aged 16-21 years). RESULTS: Of the cohort, 198 (20%) had used cannabis prior to the age of 16 years. Among this high-risk group, rates of dependence were high with 21.7% meeting DSM-IV criteria for cannabis dependence by the age of 21 years. There were clear tendencies for rates of cannabis dependence to increase with increasing reports of positive responses to early cannabis use: those reporting 5 positive responses had odds of cannabis dependence that were 28.5 (95% confidence interval, 6.3-133.8) times higher than those not reporting positive reactions to cannabis. The association held (odds ratio, 23.4; 95% confidence interval, 4.0-135.9) after control for potentially confounding factors including the extent of use of cannabis prior to age 16 years. The extent of early negative reactions to cannabis was unrelated to later cannabis dependence. CONCLUSIONS: Early subjective responses to cannabis are prognostic of later cannabis dependence. These findings may suggest the presence of genetically mediated individual differences in early responsiveness to cannabis. Clinicians should be aware that young people who report positive reactions to early use of cannabis are at increased risks of later cannabis dependence. PMID- 14557150 TI - Community reinforcement therapy for cocaine-dependent outpatients. AB - OBJECTIVE: To examine the contributions of community reinforcement therapy to outcome in the community reinforcement approach (CRA) + vouchers outpatient treatment for cocaine dependence. METHODS: One hundred cocaine-dependent outpatients were randomly assigned to one of 2 treatment conditions: CRA + vouchers or vouchers only. All patients earned incentives in the form of vouchers exchangeable for retail items contingent on cocaine-free urinalysis results during treatment weeks 1 to 12. Incentives were combined with a 24-week course of CRA therapy designed to promote healthy lifestyle changes in the CRA + vouchers condition, while incentives represented the primary treatment in the vouchers only condition. Patient drug use and psychosocial functioning were assessed at intake and at least every 3 months for 2 years after treatment entry. RESULTS: Patients treated with CRA + vouchers were retained better in treatment, used cocaine at a lower frequency during treatment but not follow-up, and reported a lower frequency of drinking to intoxication during treatment and follow-up compared with patients treated with vouchers only. Patients treated with CRA + vouchers also reported a higher frequency of days of paid employment during treatment and the initial 6 months of follow-up, decreased depressive symptoms during treatment only, and fewer hospitalizations and legal problems during follow-up. CONCLUSIONS: Combining CRA with vouchers had therapeutic effects on substance abuse and psychosocial functioning during treatment and posttreatment follow-up in cocaine-dependent outpatients, although effects on cocaine use appear to be limited to the treatment period. PMID- 14557152 TI - Hearts and minds and child restraints in airplanes. PMID- 14557154 TI - Day care for preschool children. PMID- 14557156 TI - Is epinephrine efficacious in the treatment of bronchiolitis? PMID- 14557155 TI - A meta-analysis of randomized controlled trials evaluating the efficacy of epinephrine for the treatment of acute viral bronchiolitis. AB - BACKGROUND: Controversy exists surrounding the use of bronchodilators for bronchiolitis. Epinephrine hydrochloride is being used with increasing frequency in this group; however, its efficacy has not been systematically reviewed. OBJECTIVE: To systematically review randomized controlled trials comparing inhaled or systemic epinephrine vs placebo or other bronchodilators. DATA SOURCES: MEDLINE, EMBASE, the Cochrane Central Register of Controlled Trials, primary authors, and reference lists. STUDY SELECTION: Studies were included if they (1) were randomized, controlled trials; (2) involved children 2 years or younger with bronchiolitis; and (3) presented quantitative outcomes. DATA EXTRACTION: Two reviewers independently extracted data and assessed study quality. DATA SYNTHESIS: We included 14 studies (7 inpatient, 6 outpatient, and 1 patient status unknown). Thirteen of forty-five comparisons were significant. Among outpatients, results favored epinephrine compared with placebo for clinical score at 60 minutes (standardized mean difference [SMD], -0.81; 95% confidence interval [CI], -1.56 to -0.07), oxygen saturation at 30 minutes (weighted mean difference [WMD], 2.79; 95% CI, 1.50-4.08), respiratory rate at 30 minutes (WMD, 4.54; 95% CI, -8.89 to -0.19), and improvement (odds ratio, 25.06; 95% CI, 4.95 126.91); among inpatients, for clinical score at 60 minutes (SMD, -0.52; 95% CI, 1.00 to -0.03). Among outpatients, results favored epinephrine compared with albuterol sulfate (salbutamol) for oxygen saturation at 60 minutes (WMD, 1.91; 95% CI, 0.38-3.44), heart rate at 90 minutes (WMD, -14.00; 95% CI, -22.95 to 5.05), respiratory rate at 60 minutes (WMD, -7.76; 95% CI, -11.35 to -4.17), and improvement (odds ratio, 4.51; 95% CI, 1.93-10.53); among inpatients, respiratory rate at 30 minutes (WMD, -5.12; 95% CI, -6.83 to -3.41). CONCLUSIONS: Epinephrine may be favorable compared with placebo and albuterol for short-term benefits among outpatients. There is insufficient evidence to support the use of epinephrine among inpatients. Large, multicentered trials are required before routine use among outpatients can be strongly recommended. PMID- 14557157 TI - Effects and costs of requiring child-restraint systems for young children traveling on commercial airplanes. AB - CONTEXT: The US Federal Aviation Administration is planning a new regulation requiring children younger than 2 years to ride in approved child-restraint seats on airplanes. OBJECTIVES: To estimate the annual number of child air crash deaths that might be prevented by the proposed regulation, the threshold proportion of families switching from air to car travel above which the risks of the policy would exceed its benefits, and the cost per death prevented. DESIGN: Risk and economic analyses. RESULTS: Child-restraint seat use could prevent about 0.4 child air crash deaths per year in the United States. Increased deaths as a result of car travel could exceed deaths prevented by restraint seat use if the proportion of families switching from air to car travel exceeded about 5% to 10%. The estimate for this proportion varied with assumptions about trip distance, driver characteristics, and the effectiveness of child-restraint seats but is unlikely to exceed 15%. Assuming no increase in car travel, for each dollar increase in the cost of implementing the regulation per round trip per family, the cost per death prevented would increase by about $6.4 million. CONCLUSIONS: Unless space for young children in restraint seats can be provided at low cost to families, with little or no diversion to automobile travel, a policy requiring restraint seat use could cause a net increase in deaths. Even excluding this possibility, the cost of the proposed policy per death prevented is high. PMID- 14557158 TI - Physician variation in test ordering in the management of gastroenteritis in children. AB - OBJECTIVES: To describe the variation among physicians in test ordering when caring for children with gastroenteritis and to explore the effect of hospital charge information on such variation. DESIGN: Prospective, nonmasked, observational study and controlled trial of price information. SETTING: Urban, university-affiliated pediatric emergency department. PARTICIPANTS: Pediatric emergency medicine faculty (n = 10) and fellows (n = 5). METHODS: Test-ordering practices were reviewed during 3 periods: control, intervention, and washout. During the intervention period, test charge information was placed on patients' emergency department records. Telephone contact with families was initiated 7 days after care. RESULTS: We included 3198 visits. Individual physician mean test charges varied more than 2-fold during the control period (mean, 127 US dollars; range, 82 US dollars-185 US dollars). Based on their test charges (control period), physicians were assigned to the "high" (n = 8) or "low" (n = 7) test user group. Differences in mean charges in high vs low test users during the control period (144 US dollars vs 112 US dollars) persisted in the intervention period (80 US dollars vs 52 US dollars; Mann-Whitney P =.01), as did rates of intravenous fluid use (20% vs 14% in both periods). Among the lowest-acuity patients, low test users exhibited greater price sensitivity (vs high users). Patients treated by low test users did not differ in improved condition (82% vs 86%) or family satisfaction (93% vs 92%); they had more unscheduled follow-up (25% vs 17%; P<.01), but were no more often admitted (5% vs 3%; P =.11). CONCLUSIONS: Physicians varied in resource use when treating children with gastroenteritis. High and low test users were sensitive to price information. This intervention did not seem to compromise patient outcome. PMID- 14557159 TI - Depression in Medicaid-covered youth: differences by race and ethnicity. AB - BACKGROUND: Racial disparities have previously been noted in antidepressant use among Medicaid-covered youth. OBJECTIVE: To determine if racial and ethnic differences are due to dissimilarity in the prevalence of diagnosed depression or disparate treatment patterns. METHODS: Claims were examined for 192 441 youth (5 18 years old) who had been continuously enrolled in Medicaid from July 1, 1997, to December 31, 1998. Diagnosed depression was defined as having been assigned an International Classification of Diseases, Ninth Revision code for a depressive disorder in a medical claim. Logistic regression methods were used to evaluate the association between race/ethnicity and (1) depression diagnosis and (2) depression treatment in the 6 months following a new episode of diagnosed depression. All analyses were controlled for age, sex, and rural or urban residence. RESULTS: Two percent of the total sample had a depression diagnosis, 25% of which were new episodes of depression. Compared with white youth, Hispanic (odds ratio [OR], 0.51; 95% confidence interval [CI], 0.46-0.57), Asian/Pacific Islander (OR, 0.16; 95% CI, 0.12-0.21), and black (OR, 0.31; 95% CI, 0.26-0.37) youth were less likely to have a depression diagnosis. Following a new diagnosis, Native American (OR, 0.29; 95% CI, 0.18-0.46) and Hispanic (OR, 0.42; 95% CI, 0.30-0.61) youth were less likely than white youth to have received an antidepressant or a mental health specialty visit. CONCLUSIONS: Racial and ethnic disparities exist in both the prevalence and treatment of diagnosed depression. Future studies should examine underlying reasons for these disparities and how they affect the quality of care for depressed Medicaid-covered youth. PMID- 14557160 TI - Prediction of resource use during acute pediatric illnesses. AB - BACKGROUND: Significant resources are used for acute illnesses in children. Identifying predictors of resource use can focus interventions to reduce this use. OBJECTIVE: To determine the relative effects of maternal, infant, social milieu, and demographic characteristics, the mother-child interaction, and perception of illness severity on the use of resources during acute illnesses in children. DESIGN: At the 2-week and 6-, 15-, and 24-month well-child care visits of a cohort of mother-infant dyads, the mother-well-child interaction was assessed by using the Biringen Emotional Availability Scales, and data were gathered regarding maternal depression and sense of competence, infant temperament, maternal social support, the home environment, and demographic characteristics. At each of the cohort's 1983 ill-child care visits during 30 months of follow-up, the mother-ill-child interaction was assessed by using the Emotional Availability Scales, and mothers and pediatricians independently assessed illness severity using the Acute Illness Observation Scales. Resources used during the illnesses-over-the-counter and prescription medications, tests, hospitalizations, follow-up visits, and the emergency department-were assessed. SETTING: A hospital primary care center and an urban and a suburban private practice. Patients Between February 1, 1995, and March 30, 1998, a consecutive sample of 380 dyads were asked to enroll at the 2-week well-child care visit; 316 (83.2%) consented, and complete data were available for analysis of 243 dyads. MAIN OUTCOME MEASURES: A path analytic framework using a structural equation model assessed the presence and strength of predictive relationships between demographic, maternal, infant, and social milieu data, the Biringen Emotional Availability Scales, and the Acute Illness Observation Scales and the main outcome measure, resource use. RESULTS: Three variables predicted greater mean resource use during each acute illness episode: a less optimal mother-child interaction (beta = -.53), lower scores for parenting sense of competence (beta = -.26), and greater perception of illness severity by mothers (beta =.33). By using the coefficient of determination (R2), these 3 predictors account for 55% of the reliable variance in resource use during acute illnesses. CONCLUSION: The quality of the mother-child interaction, maternal sense of competence, and maternal assessment of severity of the illness are major predictors of resource use during acute pediatric illnesses, and should be important foci of interventions to reduce resource use. PMID- 14557161 TI - Trends in psychotropic medication costs for children and adolescents, 1997-2000. AB - OBJECTIVE: To examine trends in psychotropic medication utilization and costs for children and adolescents between January 1, 1997, and December 31, 2000. METHODS: Pharmacy claims were analyzed for mental health users 17 years and younger (N = 83 039) from a national database covering 1.74 million privately insured youths. Utilization rates and costs for dispensed medications were compared across psychotropic drug categories and individual agents over time. RESULTS: Overall use of psychotropic drugs increased from 59.5% of mental health outpatients in 1997 (a 1-year prevalence of 28.7 per 1000) to 62.3% in 2000 (33.7 per 1000), a 4.7% increase. The largest changes in utilization were seen for atypical antipsychotics (138.4%), atypical antidepressants (42.8%), and selective serotonin reuptake inhibitors (18.8%). The average prescription price increased by 17.6% (7.90 US dollars per prescription), a change in turn attributed to a shift toward costlier medications within the same category (55.1% of the increase, or 4.35 US dollars) and to pure inflation (44.9% of the increase, or 3.55 US dollars; P for trend <.001 for all comparisons). Almost half (46.7%) of the 2.7 million US dollars gross sales differential was accounted for by only 3 of the 39 drugs identified (amphetamine compound, risperidone, and sertraline), and 75% was accounted for by 7 drugs (the previous 3 and bupropion, paroxetine, venlafaxine, and citalopram). CONCLUSIONS: Psychotropic drug expenditure increases during the late 1990s resulted from more youths being prescribed drugs, a preference for newer and costlier medications, and the net effects of inflation. The impact of managed care and pharmaceutical marketing effects on these trends warrants further study. PMID- 14557162 TI - Traumatic low attenuation subdural fluid collections in children younger than 3 years. AB - OBJECTIVE: To determine the time course for the development of posttraumatic nonhemic subdural fluid collections in infants and young children. DESIGN: Retrospective consecutive case series during 16 years. Patients Fifty-five head trauma patients younger than 3 years with low attenuation subdural fluid on computed tomography. MAIN OUTCOME MEASURE: Time after head trauma when low attenuation fluid first becomes visible. SETTING: Regional pediatric medical center. RESULTS: The initial visualization of low attenuation subdural fluid was within 4 days of the trauma for 44 of the patients. The mean +/- SD size of the subdural fluid collections when first identified was 4.6 +/- 2.0 mm (range, 2-12 mm), and the maximum observed size was 7.7 +/- 3.5 mm (range, 3-21 mm). The mean +/- SD time after injury until the maximum observed size was 16 +/- 18 days (range, 0-87 days). Low attenuation subdural fluid and high attenuation intracranial hemorrhage coexisted on at least 1 computed tomographic study during the first week after the trauma in 42 (81%) of the 52 patients with hemorrhage. CONCLUSION: Low attenuation subdural fluid collections (distinct from clotted blood) in infants and young children with head injuries most often develop during the first week after the traumatic event. PMID- 14557163 TI - Abusive head injury as a cause of apparent life-threatening events in infancy. AB - BACKGROUND: An apparent life-threatening event (ALTE) refers to the sudden occurrence of a breathing abnormality, color change, or alteration in muscle tone or mental status in an infant. Several patients with ALTEs admitted to our institution were found to have sustained abusive head injuries. OBJECTIVE: To systematically examine the possible causes of ALTEs and their relative frequencies. STUDY DESIGN: Prospective consecutive case series of 243 infants younger than 12 months admitted to a tertiary care academic medical center for evaluation of an ALTE during a 32-month interval. RESULTS: Thirty-five different causes of ALTEs were identified. Six subjects (2.5%) were diagnosed as having abusive head injuries, or 1 admission every 5 months. Three patients died in the hospital, 2 of whom were diagnosed as having abusive head injuries. CONCLUSIONS: A wide spectrum of diseases and disorders can precipitate an ALTE. Among them, abusive head injury, a recently recognized cause, occurs frequently enough to obligate its inclusion in the differential diagnosis. An ophthalmologic evaluation with dilated fundus examination and cranial imaging should therefore be considered early in the investigation unless another cause becomes apparent soon after admission. PMID- 14557165 TI - The impact of the hepatitis B virus vaccine on the incidence of hepatitis B virus associated membranous nephropathy. AB - BACKGROUND: The hepatitis B virus (HBV) vaccine has resulted in a decline in the incidence of HBV carriage and hepatocellular carcinoma in southeast Asia. Vaccine efficacy in Africa has not been adequately assessed. OBJECTIVE: To report on the impact of HBV vaccination in South Africa on HBV-associated membranous nephropathy (MN) over 6 years. METHODS: King Edward VIII Hospital in Durban is the only tertiary referral center for the province of KwaZulu-Natal for children with renal diseases. The HBV vaccine was introduced into the South African Expanded Programme on Immunisation on April 1, 1995; vaccine coverage rates between April 1, 1995, and December 31, 2001, for children for the first, second, and third doses were 85.4%, 78.2%, and 62.0%, respectively. Hepatitis B virus status was determined using a radioimmunoassay (January 1, 1984-March 31, 1991) or an enzyme-linked immunosorbent assay. Membranous nephropathy was confirmed by the results of a renal biopsy. The hospital average annual incidence of HBV associated MN was compared before and after vaccination and according to age groups. RESULTS: Between January 31, 1984, and December 31, 2001, there were 119 children with HBV-associated MN; their mean age was 7 years (range, 1-14 years), and 101 (84.9%) were males. The average annual rate ratio per 105 child population was 0.25. The overall incidence rate ratio showed a significant decrease from January 1, 2000, to December 31, 2001, compared with the preimmunization period (January 1, 1984-December 31, 1994) (incidence rate ratio, 0.12; 95% confidence interval, 0.03-0.50). Children from birth to the age of 4 years experienced no disease after 1998. Children aged 5 to 10 years showed a significant decrease in 2000-2001 compared with the prevaccination years (incidence rate ratio, 0.19; 95% confidence interval, 0.05-0.80). CONCLUSION: The HBV vaccine, even at low coverage for the full South African Expanded Programme on Immunisation schedule, reduced the hospital incidence of HBV-associated MN over 6 years. PMID- 14557164 TI - Lake-associated outbreak of Escherichia coli O157:H7 in Clark County, Washington, August 1999. AB - CONTEXT: Escherichia coli O157:H7, one of hundreds of strains of the gram negative bacterium E coli, has been implicated in numerous lake-borne outbreaks of infection during the past decade. In August 1999, several children who later became ill with E coli O157:H7 infection reported swimming in a lake in Clark County, Washington. The lake was closed and an investigation begun. OBJECTIVES: To identify the source of the outbreak and determine risk factors for infection with E coli O157:H7.Design, Setting, and Patients Two case-control studies were performed among residents of and visitors to Clark County in August 1999 by using community and campground-registrant control subjects. Main Outcome Measure Risk factors for infection with E coli O157:H7 among Clark County residents or visitors. RESULTS: We identified 37 case patients (including 29 primary-case patients) with a median age of 5 years (age range, 1-14 years for primary-case patients). Eight children were hospitalized, 3 with hemolytic uremic syndrome; none died. With analysis restricted to primary-case patients, illness was strongly associated with swimming in the lake (18 of 18 case patients vs 1 of 18 neighborhood-matched and age-matched control subjects; matched odds ratio undefined; P<.001). All primary-case patients were children younger than 15 years who swam in the lake. Illness was associated with placing the head underwater, getting lake water in the mouth, or swallowing lake water (26 of 27 case patients vs 43 of 62 control subjects; matched odds ratio = 11.5; P =.005). Cultures of lake water yielded E coli O157:H7 that matched the outbreak strain according to results of pulsed-field gel electrophoresis. CONCLUSIONS: To date, this is one of the largest documented outbreaks of E coli O157:H7 infection associated with unchlorinated recreational water and represents the first outbreak in which the strain was isolated from lake water. Guidelines are needed to decrease the risk of enteric illness associated with swimming in recreational lakes. PMID- 14557166 TI - Odorants in breast milk. PMID- 14557167 TI - In the hands of children: fatal car, van, and truck crashes involving drivers aged 7 through 14 years. PMID- 14557168 TI - Subspecialty care use rates: reverse causation and generalizability issues. PMID- 14557169 TI - Pediatric referral patterns. PMID- 14557170 TI - Evaluation of the clinical course of dry eye syndrome. AB - OBJECTIVE: To assess subjective symptoms, tear function factors, and ocular surface morphology in the clinical course of patients with dry eye syndrome under treatment within an observation period of up to 8 years. METHODS: In 97 patients (78 women and 19 men) with ocular discomfort, a clinical diagnosis of dry eye syndrome was made based on typical symptoms and a reduced tear film breakup time of less than 10 seconds. Subsequent evaluations revealed a diagnosis of aqueous tear deficiency in 9 patients, meibomian gland dysfunction in 32 patients, and aqueous tear deficiency combined with meibomian gland dysfunction in 30 patients, aqueous tear deficiency associated with Sjogren syndrome in 12 patients, and aqueous tear deficiency and meibomian gland dysfunction associated with Sjogren syndrome in 14 patients. Follow-up assessments were performed 12 to 94 months (mean follow-up, 40 months) after the initial diagnosis. MAIN OUTCOME MEASURES: In different subgroups of dry eye tear film breakup time, Schirmer test without local anesthesia (Schirmer I), fluorescein and rose bengal staining, impression cytology, as well as subjective dry eye symptoms and frequency of tear substitute application were compared at baseline and after a follow-up of 1 to 8 years (mean, 3.3 years). RESULTS: At baseline, tear film function and ocular surface test results found more pathologic abnormalities and more severe subjective symptoms in patients with aqueous tear deficiency associated with Sjogren syndrome and aqueous tear deficiency and meibomian gland dysfunction associated with Sjogren syndrome compared with the other groups who had dry eye syndrome. No differences in frequency of tear substitute application were observed. At follow up, tear breakup time, Schirmer I test results, and corneal fluorescein staining improved compared with baseline values, whereas rose bengal staining and impression cytology of the conjunctival surface remained almost unchanged. Subjective symptoms and frequency of artificial tear application were reduced. CONCLUSIONS: Within the observation period of up to 8 years, the dry eye syndrome improved or stabilized under appropriate treatment. Although no patient was completely cured, subjective reports as well as frequency of artificial tear application were reduced. PMID- 14557171 TI - Amniotic membrane with living related corneal limbal/conjunctival allograft for ocular surface reconstruction in Stevens-Johnson syndrome. AB - OBJECTIVE: To report the use of preserved amniotic membrane and living related corneal limbal/conjunctival transplantation in total limbal stem cell and conjunctival deficiency secondary to severe Stevens-Johnson syndrome. DESIGN: Prospective, noncomparative, interventional case series. PATIENTS AND METHODS: Ten eyes of 10 patients with total limbal stem cell and conjunctival deficiency secondary to Stevens-Johnson syndrome underwent excision of cicatricial tissue followed by amniotic membrane and living related corneal limbal/conjunctival transplantation. MAIN OUTCOME MEASURES: Reconstruction of corneal epithelium (clear appearance without epithelial defect, normal fluorescein permeability, and the absence of conjunctiva-derived goblet cells on impression cytologic testing), decrease in corneal vascularization, and improvement in visual acuity. RESULTS: During a mean follow-up of 16.7 months, satisfactory ocular surface reconstruction was obtained in 2 eyes (20%), with reduced inflammation and vascularization and a mean epithelialization time of 3 weeks. Surgical failure was observed in 4 cases (40%) and complications (infection) in 4 cases (40%). Visual acuity improved in 4 eyes (40%), remained stable in 5 eyes (50%), and decreased in 1 eye (10%). CONCLUSIONS: Amniotic membrane and living related corneal limbal/conjunctival transplantation were successful in 20% of severe cases of total limbal stem cell and conjunctival deficiency secondary to Stevens Johnson syndrome. A high proportion of postoperative complications, in particular, infection, seemed to jeopardize a favorable outcome. PMID- 14557172 TI - Evaluation of minimally invasive therapies and rationale for a prospective randomized trial to evaluate selective intra-arterial lysis for clinically complete central retinal artery occlusion. AB - OBJECTIVES: To determine the effect of commonly used minimally invasive treatments for clinically complete nonarteritic central retinal artery occlusion (CRAO) and design a prospective randomized trial to evaluate selective intra arterial lysis for this condition. METHODS: In this retrospective noncomparative case series, all medical records of patients with a diagnosis of CRAO treated at the Department of Ophthalmology, Ludwig-Maximilians-Universitat, Munich, Germany, from 1994 through 1999 were reviewed for treatments administered and course of visual acuity. MAIN OUTCOME MEASURES: Best-corrected visual acuity (BCVA) at initial and last visit. RESULTS: We identified 102 patient medical records; 71 were suitable for further analysis. Forty-four (62%) of the 71 patients included were treated with oral acetylsalicylate; 44 (62%), with oral acetazolamide; 32 (45%), with ocular massage; 22 (31%), with isovolemic hemodilution; 19 (27%), with oral pentoxifylline; 8 (11%), with topical beta-blocker; 6 (8%), with paracentesis of the anterior chamber; 4 (6%), with subcutaneous heparin. A mean +/- SD number of treatments of 2.5 +/- 1.4 was administered per patient, and BCVA increased by a mean +/- SD number of Snellen lines of 0.7 +/- 2.8. The BCVA in 11 patients (15%) increased by 3 or more lines. Multivariate stepwise regression did not reveal any single or combination treatment as a significant factor for improvement in BCVA. Patient age and duration of visual impairment before initial examination were not significant predictors of final BCVA. CONCLUSIONS: Commonly used minimally invasive treatments of CRAO do not improve the natural course of the disease. A prospective trial by the European Assessment Group for Lysis in the Eye is under way to evaluate selective intra-arterial lysis, and in this trial some of these minimally invasive treatments are used in the control group. PMID- 14557173 TI - Chorioretinal anastomosis after radial optic neurotomy for central retinal vein occlusion. AB - OBJECTIVES: To evaluate the incidence of chorioretinal anastomosis after radial optic neurotomy and to determine its effect on visual acuity and foveal thickness in patients with central retinal vein occlusion. METHODS: We conducted a prospective, uncontrolled, interventional study of 14 patients with preoperative visual acuities below 20/125. Pars plana vitrectomy and radial optic neurotomy were performed. Fluorescein angiography and optical coherence tomography were used to monitor the evolution of macular edema. RESULTS: All patients underwent radial optic neurotomy with no major complications. Eight patients (57.1%) gained 1 or more lines of visual acuity while the visual acuity of 6 patients (42.9%) improved by 2 or more lines (mean visual acuity, 20/80; P<.001) (mean visual acuity gain, 3 lines). The decrease in macular thickness was shown to be statistically significant (P<.001) (median, 282 microm). Retinochoroidal shunts developed in 6 eyes (42.9%) at the site of the radial optic neurotomy. MAIN OUTCOME MEASURES: Improvement in visual acuity and a decrease in foveal thickness seen on optical coherence tomography. CONCLUSIONS: Surgical decompression of central retinal vein occlusion via radial optic neurotomy seems to be a promising technique that improves or at least stabilizes the course of severe central retinal vein occlusion. Improvement may occur because of optic nerve decompression, vitrectomy, and by inducing new chorioretinal shunts that drain retinal circulation to the choroid and accelerate resolution of retinal edema. PMID- 14557174 TI - Polypoidal choroidal vasculopathy: incidence, demographic features, and clinical characteristics. AB - OBJECTIVE: To clarify the incidence, demographic features, and clinical characteristics of polypoidal choroidal vasculopathy (PCV) in Japanese patients. METHODS: Consecutive patients with presumed neovascular age-related macular degeneration (AMD) who met the eligibility criteria were examined between January 1, 1999, and October 31, 2001. All patients underwent complete ophthalmologic examination and fluorescein and indocyanine green angiography. RESULTS: Among 471 eyes of 418 patients who met the criteria, 110 eyes (23%) of 100 patients were diagnosed as having PCV and 361 eyes (77%) of 318 patients as having neovascular AMD. Mean age of patients with PCV was 68.4 years, with a male preponderance (63% of patients); involvement was mostly unilateral (90% of patients), and polypoidal vascular lesions were located mainly in the macula (85% of eyes). Retinal manifestations of PCV were characterized by serous macular detachment (52% of eyes), submacular hemorrhage (30% of eyes), and retinal pigment epithelium degeneration (10% of eyes). There were few subretinal fibrovascular proliferations (7% of eyes). Mean visual acuity was 0.31 in eyes with PCV and 0.18 in eyes with AMD. The incidence of severe visual loss (0.2 or worse) was 35% in PCV and 53% in AMD. CONCLUSIONS: The incidence of PCV in Japanese patients is high, and the incidence and demographic features vary in different ethnic groups. The clinical manifestations of PCV and AMD resemble each other; however, PCV is characterized by low incidence of subretinal fibrovascular proliferation, slow progression of vascular abnormality, and minimal association with conventional choroidal neovascularization. These factors seem to lead to a more favorable visual outcome in PCV compared with neovascular AMD. PMID- 14557175 TI - Prognostic value of the disodium phosphate 32P uptake test in uveal melanoma: a long-term study. AB - OBJECTIVE: To evaluate whether nuclear activity as measured by the disodium phosphate 32P (32P) uptake test for uveal melanoma is of prognostic value and corresponds to known prognostic factors. METHODS: A retrospective analysis of 121 patients with choroidal and/or ciliary body melanoma, tested with the 32P uptake test before enucleation between January 1, 1973, and December 31, 1976, at the Leiden University Medical Center. We obtained the 25-year follow-up information of this group of patients and compared the 32P test results and histopathological variables with the long-term survival rates. RESULTS: The cumulative 5-, 10-, and 20-year survival for melanoma-related death was 81.4%, 73.3%, and 63.9%, respectively. The results of the 32P uptake test were not significantly correlated with survival (P =.35). Of all prognostic factors under study, tumor diameter, cell type, and mitotic count were identified as the most important prognostic markers for uveal melanoma in this group. CONCLUSIONS: The 32P isotope uptake test has no prognostic value for uveal melanoma. Moreover, the results of this study indicate that it is unlikely that cell activity as determined by 32P uptake involves mitotic activity of the tumor. PMID- 14557177 TI - Nonmechanical Q-switched erbium:YAG laser trephination for penetrating keratoplasty: experimental study on human donor corneas. AB - OBJECTIVE: To assess the alterations in human donor corneal tissue induced by Q switched erbium (Er):YAG laser corneal trephination. METHODS: Thirty human corneoscleral donor buttons unsuitable for transplantation were placed in an artificial chamber on an automated rotation device. Corneas were trephined with a Q-switched Er:YAG laser (wavelength, 2.94 microm; pulse duration, 400 nanoseconds) along (donor and recipient) aluminum silicate (ceramic) open masks. A spot diameter of 0.65 mm, energy setting of 50 mJ/pulse, and repetition rate of 5 Hz were used. Corneal thermal damage and cut regularity were quantitatively assessed in 24 corneas processed for light microscopy and by transmission and scanning electron microscopy. RESULTS: The stromal thermal damage was the highest (mean [SD], 8.0 [2.7] microm) at a 150-microm cut depth and decreased downward. Cut regularity was very good and did not significantly differ between donors and recipients. Scanning electron microscopy confirmed that the cuts were highly regular; transmission electron microscopy revealed 2 distinctive subzones within the stromal thermal damage zone. CONCLUSIONS: Thermal damage induced by Q switched Er:YAG nonmechanical corneal trephination was low, and the regularity of the cuts was very good. CLINICAL RELEVANCE: The Q-switched Er:YAG laser may have the potential to become an alternative to the excimer laser for nonmechanical penetrating keratoplasty. PMID- 14557176 TI - Assessment of multifocal electroretinogram abnormalities and their relation to morphologic characteristics in patients with large drusen. AB - OBJECTIVES: To determine the extent of functional changes in the first-order kernel multifocal electroretinogram (mfERG) responses in patients with large drusen by means of a localized analysis and to determine correlations between mfERG responses and morphologic changes. METHODS: Thirty-one eyes from 20 patients ages 58 to 84 years with large drusen (> or =5 drusen > or =63 microm diameter) were studied. The mfERGs were recorded with a stimulus of 103 hexagons and a flash intensity of 2.67 candela (cd).s-1.m-2. Each of the 103 single first order kernel mfERG responses was analyzed and compared with those of age-matched healthy control subjects. Imaging studies, including color stereo fundus photography, red-free fundus photography, and fluorescein angiography, were performed in all patients, and morphologic changes (drusen in red-free fundus photography, staining or window defect in fluorescein angiography) were determined with a digital measurement tool. The mfERG responses were correlated to areas with and without morphologic changes. RESULTS: Reduced responses were found in 10.0% (scalar products) and 4.0% (response densities) and delayed implicit times in 13.8% (N1), 18.9% (P1), and 23.8% (N2) of all mfERGs. Abnormal mfERG responses extended up to 25 degrees in radius. Significant morphologic functional relations were detected in only a few patients. Abnormal mfERG variables were present in areas without morphologic changes. CONCLUSIONS: Patients with large drusen exhibit functional changes in the cone-driven pathways evaluated by the mfERG, indexed particularly by implicit times. Morphologically visible changes do not predict retinal function. Large drusen are associated with a more general retinal dysfunction. PMID- 14557178 TI - Cytotoxicity of indocyanine green on retinal pigment epithelium: implications for macular hole surgery. AB - OBJECTIVE: To evaluate the potential cytotoxic effects of indocyanine green (ICG) on cultured human retinal pigment epithelium (RPE) and the resultant implications for macular hole surgery. METHODS: Human RPE cells were exposed to ICG in concentrations from 0.001 to 5 mg/mL. The exposure duration ranged from 5 minutes to 3 hours. Light microscopy, MTS viability assay, and calcein AM-ethidium homodimer 1 staining were used to evaluate the cytotoxic effects of ICG. RESULTS: The RPE cells incubated with up to 5 mg/mL of ICG for 5 minutes or less exhibited no morphologic change and no significant decrease in dehydrogenase activity. When RPE cells were exposed to 5 mg/mL of ICG for 10 minutes, 1 mg/mL of ICG for 20 minutes, or 0.01 mg/mL of ICG for 3 hours, cell morphologic features were altered, mitochondrial dehydrogenase activity decreased, and some cells were necrotic. CONCLUSIONS: Indocyanine green caused cytotoxicity in cultured human RPE in a dose- and time-dependent manner. Cell death occurred by necrosis. CLINICAL RELEVANCE: Exposure of RPE cells to ICG concentrations up to 5 mg/mL for 5 minutes or less was not injurious; prolonged exposure to a low ICG concentration was toxic. Since ICG may be retained in the vitreous cavity for a lengthy period, thorough washout of ICG during macular hole surgery is required. PMID- 14557179 TI - A population-based study of the refractive outcome in 10-year-old preterm and full-term children. AB - OBJECTIVE: To evaluate the refractive outcome in 10-year-old prematurely born children and in full-term control children. METHODS: Retinoscopy during cycloplegia was performed in 213 prematurely born children from a previous population-based study on the incidence of retinopathy of prematurity and in 217 children born at term. The spherical equivalent, astigmatism, anisometropia, and significant refractive errors (defined as hypermetropia >3 diopters [D], myopia < or =-1 D, astigmatism > or =1 D in 1 or both eyes, and/or anisometropia > or =1 D) were analyzed. RESULTS: Significant refractive errors were found in 29.6% of the prematurely born and in 7.8% of the full-term children. Prematurely born children had higher prevalences of hypermetropia of more than 3 D, myopia of -1 D or less, astigmatism of 1 D or more, and anisometropia of 1 D or more than those born at term. In the preterm group, the cryotreated children had the greatest risk of refractive errors (16 [64%] of 25 children), with higher prevalences of myopia (<0, < or =-1, or <-3 D), astigmatism (> or =1 D), and anisometropia (> or =1 D). CONCLUSIONS: Significant refractive errors were 4 times more common in 10 year-old prematurely born children than in full-term controls. Cryotreated children had the highest risk, but prematurity per se was also associated with refractive errors. Ophthalmological follow-up of prematurely born children should, therefore, also include children without retinopathy of prematurity in the neonatal period. PMID- 14557180 TI - Epidemiology of childhood ocular trauma in a northeastern Colombian region. AB - OBJECTIVE: To describe the epidemiology of ocular trauma in children 15 years and younger who underwent evaluation during a 5-year period in the emergency department of a tertiary referral center in northeastern Colombia. METHODS: We retrospectively reviewed the medical records of children 15 years and younger. Records of 393 children with 415 incidents of eye injury were included in the study, of whom 22 were initially treated for bilateral ocular trauma. RESULTS: Most patients (64.9%) were boys. The highest proportion of injuries (44.4%) occurred at home, followed by streets and roads (28.6%). Blunt (35.1%) and sharp (22.6%) objects represented the most frequent causes of trauma. Closed-globe injuries were far more frequent than open-globe injuries for boys (82.4% vs 17.6%) and girls (83.8% vs 16.2%). Of those with closed-globe injuries, 253 injuries (80.0%) registered an initial visual acuity of greater than 20/60, whereas 31 open-globe injuries (52.5%) registered an initial visual acuity of less than 20/400. Most closed-globe injuries (223 [92.1%]) did not cause any final visual impairment in the affected eye, whereas 26 open-globe injuries (55.3%) caused severe visual impairment or blindness. CONCLUSIONS: Most of the accidents reported in this study could have been avoided; the data demonstrate a clear need for primary prevention and control measures. PMID- 14557181 TI - Retinal emboli and cardiovascular disease: the Beaver Dam Eye Study. AB - OBJECTIVE: To describe the 10-year incidence of retinal emboli, associated risk factors, and relationship of retinal emboli at baseline to stroke and ischemic heart disease mortality. METHODS: The Beaver Dam Eye Study is a large (N = 4926) population-based study of persons 43 to 86 years of age at the time of the baseline examination. Retinal emboli were detected at baseline (1988-1990) and at a 5-year (1993-1995) and 10-year (1998-2000) follow-up by grading of stereoscopic 30 degrees color fundus photographs using standardized protocols. Cause-specific mortality was determined from death certificates. RESULTS: The 10-year cumulative incidence of retinal emboli was 1.5%. After adjustment for age and sex, the incidence of retinal emboli was associated with increased pulse pressure (odds ratio [OR] [fourth vs first quartile range], 2.42; 95% confidence interval [CI], 0.98-5.97; P =.03, for test of trend), higher serum total cholesterol level (OR, 2.77; 95% CI, 1.06-7.23; P =.03), higher white blood cell count (OR, 2.28; 95% CI, 1.04-4.96; P =.05), smoking status (OR [current vs never-smoker], 4.60; 95% CI, 2.08-10.16; P<.001), and a history of coronary artery bypass surgery (OR, 7.17; 95% CI, 3.18-16.18; P<.001) at baseline. After controlling for age, sex, and systemic factors, a significantly higher hazard of dying with a mention of stroke on the death certificate was found in people with retinal emboli (hazard ratio, 2.40; 95% CI, 1.16-4.99) compared with those without. CONCLUSIONS: We found associations of smoking and cardiovascular disease with the incidence of retinal emboli. Persons with retinal emboli are also at increased risk of stroke related death. PMID- 14557182 TI - A peculiar autosomal dominant macular dystrophy caused by an asparagine deletion at codon 169 in the peripherin/RDS gene. AB - OBJECTIVE: To describe the clinical and genetic findings in a family with a peculiar autosomal dominant macular dystrophy with peripheral deposits. METHODS: All family members underwent an ophthalmic examination, and their genomic DNA was screened for mutations in the human retinal degeneration slow (peripherin/RDS) and rhodopsin genes. In selected cases, fluorescein angiography and electrophysiologic testing were performed. RESULTS: The age at onset of the disease was between the third and fourth decades of life, starting with mild visual acuity loss and periods of metamorphopsia. Clinical signs included subretinal yellowish macular deposits evolving into geographic atrophy and retinal hypopigmentation and hyperpigmentation. Electroretinography demonstrated rod dysfunction, and electro-oculograms were mildly to severely disturbed. All affected members were found to carry a 3-base pair deletion affecting codon 169 of the peripherin/RDS gene. This mutation resulted in an asparagine (Asn) deletion in the peripherin/RDS protein and was not found in 155 control individuals. CONCLUSION: A deletion of Asn169 in the peripherin/RDS protein causes a peculiar form of autosomal dominant macular dystrophy in a large family from the Netherlands. CLINICAL RELEVANCE: Characterizing the phenotype and genotype in this family may, in the long term, result in a better understanding of the precise mechanism underlying this retinal degeneration. PMID- 14557184 TI - Estimates of incidence rates with longitudinal claims data. AB - OBJECTIVE: To estimate incidence rates of the 3 major chronic eye diseases- diabetic retinopathy (DR), glaucoma, and age-related macular degeneration (ARMD)- by using longitudinal claims data from Medicare. METHODS: Longitudinal cases were ascertained by using a national probability sample of Medicare beneficiaries aged 65 years and older in 1991 who initially had none of the eye diseases documented. After adjusting for death and enrollment in a health maintenance organization, claims filed by optometrists or ophthalmologists with an International Classification of Diseases, Ninth Revision, Clinical Modification code for all forms of DR, glaucoma, and ARMD were used to indicate diagnosis. RESULTS: Annual incidence rates for the 3 conditions after the first year of observation ranged from 14.3% to 17.7% (higher earlier) across an 8-year longitudinal follow-up. Incidence rates among those with diabetes mellitus for any form of DR varied between 3.8% and 6.5%, while those for glaucoma varied between 4.6% and 7.8% and those for ARMD varied between 7.5% and 9.3%. CONCLUSIONS: Longitudinal claims data after the first year provide relatively stable estimates of incidence rates on an annual basis. These estimates are comparable with those of the few population-based studies available. PMID- 14557183 TI - Autosomal dominant central areolar choroidal dystrophy and a novel Arg195Leu mutation in the peripherin/RDS gene. PMID- 14557185 TI - Ophthalmology in Sweden. PMID- 14557186 TI - Ophthalmology in Finland. PMID- 14557187 TI - Idiopathic limbal squamous metaplasia. PMID- 14557188 TI - Unilateral tuberculous conjunctivitis with tarsal necrosis. PMID- 14557189 TI - Epstein-Barr virus-associated leiomyosarcoma of the iris in a child infected with human immunodeficiency virus. PMID- 14557190 TI - Adenocarcinoma of the retinal pigment epithelium. PMID- 14557192 TI - Morgagnian cataract with an isolated posterior capsular opening. PMID- 14557191 TI - Unilateral conjunctival-corneal argyrosis simulating conjunctival melanoma. PMID- 14557193 TI - Perivenular macular whitening during acute central retinal vein occlusion. PMID- 14557194 TI - Intravitreal triamcinolone for radiation-induced macular edema. PMID- 14557196 TI - In vivo confocal microscopy and genotyping of a family with Thiel-Behnke (honeycomb) corneal dystrophy. PMID- 14557195 TI - Bilateral neuroretinitis in a 6-year-old boy with acquired toxoplasmosis. PMID- 14557197 TI - Scanning laser polarimetry and progression of visual field defects after intraocular pressure increase. PMID- 14557198 TI - Resolution of hypotony maculopathy. PMID- 14557199 TI - Macular hole and macular detachment in severe hypertensive retinochoroidopathy. PMID- 14557200 TI - The multifocal electroretinogram in acute macular neuroretinopathy. PMID- 14557201 TI - Blockage of retrograde axonal flow after retinal artery occlusion: ophthalmoscopic findings. PMID- 14557202 TI - Interface corneal epithelial cyst following epikeratoplasty. PMID- 14557203 TI - Progression of congenital ectropion uveae. PMID- 14557204 TI - Chemoreduction for retinoblastoma. PMID- 14557205 TI - Health insurance and cancer survival. PMID- 14557206 TI - Should older drivers have to prove that they are able to drive? PMID- 14557207 TI - Driver screening for older adults. PMID- 14557210 TI - Cancer survival in Kentucky and health insurance coverage. AB - BACKGROUND: Access to health insurance influences the amount and quality of health care received, which in turn is likely to be related to survival. Few studies have systematically examined cancer survival by individual level health insurance data from a state population-based cancer registry for 4 anatomic sites. METHODS: Men and women aged 18 to 99 years who were registered from 1995 to 1998 with the Kentucky Cancer Registry, Lexington, with colorectal, lung, breast, or prostate cancer were followed up through 1999. Three-year crude and relative survival proportion by 7 health insurance categories and by sex for all 4 sites were calculated. Poisson regression was used to model the risk of death (controlling for age group at diagnosis, sex, race, stage at diagnosis, and treatment) relative to expected deaths in the general population from all 4 cancers by health insurance category. RESULTS: Among patients with prostate cancer, 3-year relative survival proportion was 98% for the privately insured and 83% for the uninsured; comparable figures were 91% and 78% for patients with breast cancer; 71% and 53% for patients with colorectal cancer; and 23% and 13% for patients with lung cancer. For all 4 cancers the uninsured ranked fifth or sixth on survival, above patients with unknown insurance type or Medicaid/welfare. CONCLUSION: These findings confirm purported disparities in cancer care and point toward the need to make quality care accessible to all segments of the population. PMID- 14557211 TI - Increases in clinically severe obesity in the United States, 1986-2000. AB - BACKGROUND: We know that Americans are increasingly becoming overweight, but we do not know whether this trend applies to clinically severe obesity (>100 lbs [45 kg] overweight), which is believed to have different causes than typical weight gain. Severe obesity is more serious for an individual's health and creates different challenges for the health care system. This study estimates trends for extreme weight categories between the years 1986 and 2000. METHODS: The data come from the Behavioral Risk Factor Surveillance System. The dependent variable is weight category according to the body mass index (BMI; calculated as weight in kilograms divided by the square of height in meters) based on self-reported weight and height. Regression models adjust for changes in population characteristics and state participation. RESULTS: Between 1986 and 2000, the prevalence of a BMI (self-reported) of 40 or greater (about 100 lbs [45 kg] overweight) quadrupled from about 1 in 200 adult Americans to 1 in 50; the prevalence of a BMI of 50 or greater increased by a factor of 5, from about 1 in 2000 to 1 in 400. In contrast, obesity based on a BMI of 30 or greater roughly doubled during the same period, from about 1 in 10 to 1 in 5. CONCLUSIONS: The prevalence of clinically severe obesity is increasing much faster than obesity. The widely published trends for overweight/obesity underestimate the consequences for physician practices, hospitals, and health plans because comorbidities and resulting service use are much higher among severely obese individuals. Accommodating severely obese patients will no longer be a rare event, and providers have to prepare to treat such patients on a regular basis. PMID- 14557212 TI - Risk of mortality by histologic type of breast cancer among women aged 50 to 79 years. AB - BACKGROUND: Recent studies suggest that the use of combined estrogen and progestin hormone replacement therapy is associated with an increased risk of invasive lobular carcinoma (ILC), but that it has little association with risk of invasive ductal carcinoma (IDC). Also, the incidence rates of ILC have risen over the past 10 years while those of IDC have remained constant. Differences in survival rates by histologic types of tumor have been reported, but few of the published studies were population based or had adequate power to address this issue. METHODS: We conducted a retrospective cohort study spanning the years 1974 through 1998 using data from the 9 cancer registries that have participated in the Surveillance, Epidemiology, and End Results Program since 1974. The cohort consisted of 164 958 women aged 50 to 79 years who had been diagnosed as having 1 of 7 histologic types of invasive breast cancer. Risks of mortality due to any cause were estimated using the Cox proportional hazards model. RESULTS: Women with ILC had a risk of mortality 11% lower than women with IDC. The magnitude of this difference has increased over the past 10 years and, from 1994 through 1998, the risk of mortality was 26% lower for women with ILC. Also, the risk of mortality was between 8% and 34% lower in women with mucinous carcinoma, comedocarcinoma, or medullary, tubular, and papillary carcinomas compared with women with IDC. CONCLUSIONS: Differences in prognosis by histologic type of breast cancer were identified. The survival rate of women 50 to 79 years old who have ILC, the cancer whose histologic type is the most closely linked with the use of combined estrogen and progestin hormone replacement therapy, is more favorable than that of women with IDC and appears to be improving over time. PMID- 14557213 TI - Knowledge and attitudes regarding cardiovascular disease risk and prevention in patients with coronary or peripheral arterial disease. AB - BACKGROUND: We compared perceptions regarding risk of cardiovascular events and benefits of cardiovascular disease (CVD) risk factor reduction between patients with peripheral arterial disease (PAD), patients with coronary artery disease (CAD), and patients without atherosclerosis (no disease). METHODS: Participants with no disease (n = 142) had a normal ankle-brachial index and no clinically evident atherosclerosis (group 1). The PAD participants (n = 136) had an ankle brachial index less than 0.90 and no other clinically evident atherosclerosis (group 2). Participants with CAD (n = 70) had a normal ankle-brachial index and a history of heart disease (group 3). Participants were interviewed regarding risk of mortality, CVD, and the importance of CVD risk factor reduction for hypothetical patients with PAD and CAD. RESULTS: All groups reported that risks of myocardial infarction, stroke, and death were higher for a patient with CAD than for a patient with PAD. Group 2 was less likely than group 3 to believe that PAD is associated with an extremely high risk of stroke (13.3% vs 28.7%; P =.005) or mortality (10.9% vs 26.6%; P =.003). Group 2 was less likely than group 1 to believe that a patient with PAD has a very high risk of myocardial infarction (13.1% vs 23.8%; P =.02), stroke (13.3% vs 27.5%; P =.003), or mortality (10.9% vs 24.3%; P =.004). Compared with group 3, a smaller percentage of patients in group 2 reported that cholesterol lowering was very important in PAD (57.5% vs 75.8%; P =.005). CONCLUSIONS: Compared with other patients, those with PAD underestimated the high risk of cardiovascular events associated with PAD and the benefits of cholesterol-lowering therapy. These findings may help explain the low rates of CVD risk factor control previously reported in patients with PAD. PMID- 14557214 TI - Bone mineral density measurement and treatment for osteoporosis in older individuals with fractures: a gap in evidence-based practice guideline implementation. AB - BACKGROUND: Osteoporosis evaluation and treatment guidelines state that, because of the high risk for future fractures, a fracture in an older individual warrants initiation of pharmacological treatment or bone mineral density (BMD) measurement followed by treatment according to BMD. We compared current practice with these guidelines. METHODS: We used the electronic data systems of a health maintenance organization to collect fracture, BMD measurement, and pharmacy data for women aged 50 to 89 years and men aged 65 to 89 years who sustained a study-defined fracture during 1998 or 1999. We determined those who had BMD measurement or pharmacological treatment for osteoporosis (bisphosphonate or estrogen) during the 2 years. We compared the evaluation and treatment data with evidence-based clinical guidelines (for women) or expert consensus (for men). RESULTS: Of 70 513 members in the eligible age groups, 2804 persons sustained study-defined fractures. Overall, only 4.6% of those with fractures had treatment initiated after the fracture. Women sustained 80.7% of the study-defined fractures; 8.4% had BMD measurement and 42.4% received any treatment during the 2 years. Bone mineral density measurement and treatment frequency decrease significantly with age in women. In men, 1.5% had BMD measurement and 2.8% received any treatment. Approximately 51% (51.2%) of women and 95.5% of men in our study population were not evaluated or treated in accord with guideline or expert recommendations. CONCLUSIONS: Evaluation and treatment rates for osteoporosis in older individuals with fractures fall far below national recommendations, especially for men. Intervention strategies should be developed and evaluated to prevent refracture in older individuals with fractures. PMID- 14557215 TI - Communitywide trends in the use and outcomes associated with beta-blockers in patients with acute myocardial infarction: the Worcester Heart Attack Study. AB - BACKGROUND: Despite the benefits associated with beta-blocker therapy in patients with acute myocardial infarction (AMI), limited recent data are available describing the extent of use of this therapy and the associated hospital and long term outcomes, particularly from the perspective of a population-based study. Data are also limited about the characteristics of patients with AMI who do not receive beta-blockers. This study examines more than 2 decades of trends in the use of beta-blockers in hospitalized patients with AMI. METHODS: Communitywide study of 10,374 patients hospitalized with confirmed AMI in all metropolitan Worcester hospitals during 12 annual periods between 1975 and 1999. RESULTS: There was a marked increase in the use of beta-blockers in hospitalized patients between 1975 (11%) and 1999 (82%). Older patients, women, and patients with comorbidities were significantly less likely to be treated with beta-blockers. After controlling for other prognostic factors, patients treated with beta blockers were less likely to develop heart failure (adjusted odds ratio [OR], 0.58; 95% confidence interval [CI], 0.53-0.63), cardiogenic shock (OR, 0.46; 95% CI, 0.39-0.54), and primary ventricular fibrillation (OR, 0.84; 95% CI, 0.65 1.08) and were less likely to die (OR, 0.26; 95% CI, 0.22-0.29) during hospitalization than were patients who did not receive this therapy. Patients who used beta-blockers during hospitalization had significantly lower death rates after hospital discharge. CONCLUSIONS: The results of this observational study demonstrate encouraging trends in the use of beta-blockers in hospitalized patients with AMI and document the benefits to be gained from this treatment. PMID- 14557217 TI - Sustained benefit of a community and professional intervention to increase acute stroke therapy. AB - BACKGROUND: The ultimate test of an educational intervention is the sustainability of the effect after the intervention ceases. METHODS: The TLL Temple Foundation Stroke Project was a quasi-experimental study aimed at increasing Food and Drug Administration-approved acute stroke therapy in a nonurban community in east Texas. During the intensive community and professional intervention (phase 2), significantly more patients with acute stroke received intravenous tissue plasminogen activator (tPA) compared with the preintervention period (phase 1). In the comparison community, no change was noted. We present the results of tPA treatment in the 6 months after the intervention ended (phase 3). RESULTS: Two hundred thirty-eight patients had a validated stroke during phase 3. Among patients who experienced an ischemic stroke, 11.2% in the intervention group received intravenous tPA compared with 2.2% in phase 1 (P =.007). In the comparison group, 1.4% received intravenous tPA in phase 3 compared with 0.7% in phase 1 (P>.99). Among eligible candidates for treatment, 69.2% were treated in phase 3 in the intervention community compared with 13.6% in phase 1 (P =.002). In the comparison group, 20.0% were treated in phase 3 compared with 6.7% in phase 1 (P =.45). There was 1 protocol violation among the 9 patients treated in the intervention community in phase 3. CONCLUSIONS: There was a sustained benefit of the intervention in increasing tPA treatments in the intervention community even after cessation of the educational effort. Treatments in the control community remained few through all 3 phases of the study. A carefully planned multilevel intervention can improve community stroke treatments even in a nonurban community. PMID- 14557216 TI - CD4 T-lymphocyte recovery in individuals with advanced HIV-1 infection receiving potent antiretroviral therapy for 4 years: the Swiss HIV Cohort Study. AB - BACKGROUND: Highly active antiretroviral therapy (HAART) for human immunodeficiency virus (HIV)-1 infection allows recovery of CD4 T lymphocytes. Few studies have explored the long-term T-lymphocyte responses to HAART. METHODS: Plasma HIV-1 RNA levels and CD4 and CD8 T-lymphocyte counts were longitudinally analyzed over 4 years in 2235 participants of the Swiss HIV Cohort, commencing HAART between 1996 and 1997. The CD4 T-lymphocyte count increase, the percentage of individuals with a CD4 T-lymphocyte count of 500/microL or greater and less than 200/microL, and the determinants of CD4 T-lymphocyte recovery were evaluated in individuals treated with continuous (CONT; n = 985) and discontinuous (DISCONT; n = 1250) HAART. RESULTS: At 4 years, 69.5% of subjects (CONT, 84.5%; DISCONT, 53.6%; P<.001) showed HIV-1 RNA levels below 400 copies/mL, while the median CD4 T-lymphocyte count increased from 190/microL to 423/microL (CONT, 486/microL; DISCONT, 343/microL; P<.001). Of the 2235 participants, 38.8% (CONT, 47.7%; DISCONT, 29.4%; P<.001) reached a CD4 T-lymphocyte count of 500/microL or greater, but in 15.6%, CD4 T-lymphocyte count remained below 200/microL (CONT, 5.9%; DISCONT, 25.9%; P<.001). Larger increases in CD4 T-lymphocyte count were associated with higher baseline HIV-1 RNA, a larger percentage of undetectable HIV-1 RNA levels, lower baseline CD8 T-lymphocyte count, and younger age. Individuals reaching a CD4 T-lymphocyte count of 500/microL or greater at 4 years were characterized by higher nadir and baseline CD4 T-lymphocyte counts and a more sustained reduction of HIV-1 RNA levels. CONCLUSIONS: At 4 years, only 39% of individuals treated with HAART reached a CD4 T-lymphocyte count of 500/microL or greater, and 16% with CD4 T-lymphocyte counts less than 200/microL remained susceptible to opportunistic infections. Treatment interruptions, a poor virologic response, and older age were the major factors negatively affecting the recovery of CD4 T lymphocytes. PMID- 14557218 TI - The effect of ethnicity on survival in male veterans referred for electrocardiography and treadmill testing. AB - BACKGROUND: Ethnic differences in the relationship between access to health care and survival are difficult to define because of many confounding factors, such as socioeconomic status and baseline differences in health. Because the Veterans Affairs health care system offers health care largely without financial considerations, it provides an ideal setting in which to identify and understand ethnic differences in health outcomes. Previous studies in this area have lacked clinical and cardiovascular data with which to adjust for baseline differences in patients' health. METHODS: Data were collected from consecutive men referred for resting electrocardiography (ECG) (n = 41 087) or exercise testing (n = 6213) during 12 years. We compared ethnic differences in survival between whites, blacks, and Hispanics after considering baseline differences in age and hospitalization status. We also adjusted for electrocardiogram abnormalities and cardiac risk factors, exercise test results, and cardiovascular comorbidities. RESULTS: White patients tended to be older and had more baseline comorbidities and cardiovascular interventions when they presented for testing. White patients had increased mortality rates compared with blacks and Hispanics. In the ECG population, after adjusting for demographics and baseline electrocardiogram abnormalities, Hispanics had improved survival compared with whites and blacks. In the exercise test population, after adjusting for the same factors, as well as adjusting for the presence of cardiovascular comorbidities, cardiac risk factors, and exercise test findings, Hispanics also exhibited improved survival compared with the other 2 ethnicities. There were no differences in mortality rates between whites and blacks. CONCLUSION: Our findings demonstrate that the health care provided to veterans referred for routine ECG or exercise testing is not associated with poorer survival in ethnic minorities. PMID- 14557219 TI - Are gifts from pharmaceutical companies ethically problematic? A survey of physicians. AB - BACKGROUND: Personalized pharmaceutical marketing to physicians, including the provision of gifts and sponsorship of educational and recreational activities, raises ethical issues. We sought to determine the degree to which physicians regarded common pharmaceutical marketing activities as ethically problematic, and to compare the views of experienced physicians and physicians-in-training. METHODS: A questionnaire that included 18 scenarios portraying interactions between physicians and the pharmaceutical industry was distributed to residents and faculty members at a US medical school. RESULTS: Most marketing activities were not thought to pose major ethical problems. Respondents tended to make distinctions about the ethical appropriateness of gifts on the basis of the monetary value and type of gift. Some respondents' views would be in violation of recent professional guidelines that address interactions between physicians and pharmaceutical companies. However, some respondents were troubled by activities that are permitted by professional guidelines. The responses of residents and faculty physicians were similar. CONCLUSIONS: Despite the recent publicity about ethical problems in relationships between physicians and the pharmaceutical industry, inexperienced and experienced physicians at a single institution continue to have a rather permissive view about a variety of marketing activities. PMID- 14557220 TI - The decreasing prevalence of reversible dementias: an updated meta-analysis. AB - BACKGROUND: In 1988, 2 meta-analyses suggested that the prevalence of reversible dementia was significantly lower than had been previously estimated. It was predicted that further work would indicate an even lower rate. The present study represents an updated meta-analysis of the true prevalence of reversible dementia. METHODS: MEDLINE was searched from 1987 through 2002. References were also gleaned from pertinent articles and relevant textbooks. Data were extracted on the nature and provenance of the studies, dementia etiology, and the proportion of cases that were potentially reversible and reversed. RESULTS: Fifty articles were identified of which 39 met the study criteria, representing 7042 patients of whom 5620 (87.2%) had dementia. Patients were classified according to etiology and, where possible (in 23 [59%] of 39 studies), whether the dementia partially or completely resolved. A much higher proportion of studies than was previously the case were either community-based (31%) or observed subjects from outpatient departments (54%). Alzheimer disease was still the commonest cause of dementia (56.3%) followed by a vascular etiology (20.3%). Conditions requiring neuroimaging made up only 2.2% of cases. Potentially reversible causes were seen in 9%, and only 0.6% of dementia cases actually reversed (0.29% partially, 0.31% fully). CONCLUSIONS: The reported proportion of dementias that reverse is much lower than previously thought. While comorbidity should always be treated for its own sake and in the hope that cognitive decline may at least be delayed, the present findings have significant clinical and economic implications for the workup of dementia. PMID- 14557221 TI - Optimizing long-term cardiac management after major vascular surgery: Role of beta-blocker therapy, clinical characteristics, and dobutamine stress echocardiography to optimize long-term cardiac management after major vascular surgery. AB - BACKGROUND: Survivors of major vascular surgery are at increased risk of late cardiac complications. OBJECTIVE: To examine the cardioprotective effect of beta blockers. METHODS: A follow-up study was conducted in 1286 patients who survived surgery for at least 30 days. Patients were screened for cardiac risk factors and dobutamine stress echocardiography (DSE) results; 1034 patients (80%) underwent preoperative DSE, and 370 (29%) received beta-blockers. The main outcome measure was late cardiac death or myocardial infarction. RESULTS: Seventy-four patients (5.8%) had late cardiac events. Cardiac event rates in patients with 0, 1 to 2, and 3 or more risk factors were 1.6%, 4.7%, and 19.2%, respectively. In patients without risk factors, beta-blockers were associated with improved event-free survival (2.8% vs 0%), and DSE had no additional prognostic value. In patients with 1 to 2 risk factors, the presence of ischemia during DSE increased cardiac events from 3.9% to 9.8%. However, if patients with ischemia were treated with beta-blockers, the risk decreased to 7.2%. In patients with 3 or more risk factors, DSE and beta-blockers stratified patients into intermediate- and high risk groups. In patients without ischemia, beta-blockers reduced the cardiac event rate from 15.1% to 9.5%, whereas the cardioprotective effect was limited in patients with 3 or more risk factors and positive DSE findings. CONCLUSIONS: Long term beta-blocker use is associated with a reduction in the cardiac event rate, except for patients with 3 or more risk factors and positive findings on DSE. PMID- 14557222 TI - Aging bone and osteoporosis: strategies for preventing fractures in the elderly. AB - As the older population increases, the incidence of osteoporotic fractures is expected to dramatically rise during the next few decades. Older patients are much more susceptible to fracture at any given bone mineral density (BMD) than are younger patients because of various factors, including the quality of aging bone, which involves more than BMD. Suppression of increased bone turnover by antiresorptive therapies, even with only small changes in BMD, can reduce fracture risk, especially in the lumbar spine. Bisphosphonate treatment can significantly reduce vertebral and nonvertebral fractures, including hip fractures, even in the very elderly. Prospective analyses show that risedronate therapy consistently and significantly reduces the risk of new morphometric vertebral fractures after 1 year in postmenopausal women. Post hoc analyses report significant reductions in the risk of 1 new clinical vertebral fracture after 6 months of risedronate therapy and after 1 year of alendronate therapy. Oral raloxifene therapy and salmon calcitonin nasal spray therapy have been shown to reduce the risk of vertebral fracture after 3 and 5 years, respectively, and post hoc data show a significant reduction in clinical vertebral fracture risk at 1 year with raloxifene use. However, neither raloxifene therapy nor calcitonin therapy reduce the risk of nonvertebral and hip fractures at currently approved doses. Bisphosphonates have been shown to be safe and efficacious with 7 years' risedronate sodium and 10 years' alendronate sodium data published, and bisphosphonates reduce bone turnover and increase BMD to a greater degree than raloxifene and calcitonin, which may partly account for their nonvertebral and hip fracture reduction effect. Therefore, bisphosphonate therapy with risedronate or alendronate should be considered in patients with low BMD at the hip and in older patients with osteoporosis and osteopenia, particularly those with an existing fracture. PMID- 14557223 TI - A better alternative for estimating renal function. PMID- 14557224 TI - Understanding statistical information: the problem of numerical interpretation. PMID- 14557225 TI - Productivity loss due to irritable bowel syndrome. PMID- 14557226 TI - Hypersensitivity pneumonitis associated with hot tub use. PMID- 14557227 TI - Anticoagulation with prosthetic cardiac valves. PMID- 14557228 TI - Heart rate variability. PMID- 14557229 TI - On the maturation of mathematical modeling of endocrine systems. PMID- 14557230 TI - Intrauterine growth restriction and reduced glomerular number: role of apoptosis. PMID- 14557231 TI - Circadian rhythm regulation: a central role for the neuropeptide vasoactive intestinal polypeptide. PMID- 14557232 TI - Our fragmentary knowledge of the regulatory functions of ANG II "fragments": are we beginning to see the light? PMID- 14557233 TI - Repeated administration of the anorectic factor prolactin-releasing peptide leads to tolerance to its effects on energy homeostasis. AB - Central administration of a single dose of prolactin-releasing peptide (PrRP) causes a reduction in both fast-induced and nocturnal food intake and body weight gain. The aim of this study was to examine the effect of repeated administration of PrRP on energy homeostasis, including a measure of the expression of the mitochondrial uncoupling protein-1 (UCP-1) in brown adipose tissue. Conscious, free-feeding animals received central injections of PrRP (4 nmol icv) or vehicle. A single injection at 1000 caused a sustained hyperthermia over the 4-h test period and an increase in the expression of UCP-1 mRNA. Repeated, twice daily injection caused a reduction in body weight gain greater than that seen in pair fed animals for the first 48-72 h. After 72 h, the animals became refractory to the actions of PrRP. The pair-fed group showed a reduction in UCP-1 mRNA expression at 48 h, which was reversed by PrRP treatment. This study indicates that PrRP exerts its effects on energy homeostasis in the short-medium term by reducing food intake and increasing energy expenditure. PMID- 14557234 TI - Central leptin gene therapy fails to overcome leptin resistance associated with diet-induced obesity. AB - The objective of this study was to determine if central overexpression of leptin could overcome the leptin resistance caused by 100 days of high-fat feeding. Three-month old-F344XBN male rats were fed either control low fat chow (Chow), which provides 15% of energy as fat, or a high-fat/high-sucrose diet (HF), which provides 59% of energy as fat. Over several weeks, the HF-fed animals spontaneously split into two groups of animals: those that became obese on the HF diet (DIO) and those that did not gain extra weight on the HF diet [diet resistant (DR)]. After 100 days of HF feeding, animals were given a single intracerebroventricular injection containing 5.75E10 particles of rAAV encoding leptin (rAAV-leptin) or control virus (rAAV-con). Chow animals responded robustly to rAAV-leptin, including significant anorexia, weight loss, and lipopenia. In contrast, DIO were completely unresponsive to rAAV-leptin. DR rats responded to rAAV-leptin, but in a more variable fashion than Chow. Unlike what was observed in Chow, the anorectic response to rAAV-leptin rapidly attenuated and was no longer significant by day 14 postvector delivery. Both DIO and DR animals were found to have reduced long-form leptin receptor expression and enhanced basal P STAT-3 in the hypothalamus with respect to Chow. rAAV-leptin caused an increase in STAT3 phosphorylation and proopiomelanocortin expression in the hypothalamus and an increase in uncoupling protein-1 in brown adipose tissue in both Chow and DR animals, but failed to do so in DIO. This suggests that central overexpression of leptin is not a viable strategy to reverse diet-induced obesity. PMID- 14557235 TI - Cholecystokinin and D-fenfluramine inhibit food intake in oxytocin-deficient mice. AB - Results from previous studies indicate that oxytocin (OT)-containing neural pathways are activated in laboratory rats after systemic administration of CCK or d-fenfluramine and that centrally released OT may participate in the anorexigenic effects of these treatments. To explore the relationship between feeding behavior and OT function, the effects of CCK and d-fenfluramine on feeding and central c Fos expression were compared in wild-type (OT+/+) and OT-deficient mice (OT-/-) of C57BL/6 background. Male OT+/+ and OT-/- mice were administered saline or CCK (1, 3, or 10 microg/kg ip) after overnight food deprivation. Saline-treated OT+/+ and OT-/- mice consumed equivalent amounts of food after an overnight fast. CCK inhibited deprivation-induced food intake in a dose-dependent manner to a similar extent in both genotypes. CCK treatment also induced similar hindbrain and forebrain patterns of increased c-Fos expression in mice of both genotypes. After treatment with d-fenfluramine (10 mg/kg ip), both OT+/+ and OT-/- mice consumed significantly less food than untreated controls, with no difference between genotypes. We conclude that OT signaling pathways are unnecessary for the anorexigenic effects of systemically administered CCK and d-fenfluramine in C57BL/6 mice. PMID- 14557236 TI - Neuropeptide FF exerts pro- and anti-opioid actions in the parabrachial nucleus to modulate food intake. AB - Neurons that synthesize the morphine modulatory peptide neuropeptide FF (NPFF; Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-NH2) densely innervate the parabrachial nucleus (PBN), an area implicated in regulating food intake. We analyzed opioid-related actions of NPFF in feeding in adult male Sprague-Dawley rats. Unilateral infusion of 2 nmol/0.5 microl of the mu-opioid receptor agonist [d-Ala2,NMe Phe4,glycinol5]enkephalin (DAMGO) into the lateral PBN increased 4-h food intake from 0.7 +/- 0.1 to 3.3 +/- 0.3 g. NPFF (1.25-5.0 nmol) prevented this hyperphagic mu-opioidergic action. In rats fed after 4-h deprivation (baseline = 12.3 +/- 0.3 g/2 h), 5 nmol of NPFF did not alter and larger doses (10 and 20 nmol) actually increased food intake (+36, 54%). Twenty nanomoles also elevated intake of freely feeding rats (from 0.7 +/- 0.1 to 5.1 +/- 1.0 g/4 h). The opioid receptor blocker naloxone (10 nmol) antagonized this increase. These data reveal both pro- and anti-opioid actions of NPFF in the PBN to modulate feeding. The mechanisms for the opposite actions of low and high concentrations of this neuropeptide in parabrachial regulation of food intake remain to be determined. PMID- 14557237 TI - An orexigenic role for mu-opioid receptors in the lateral parabrachial nucleus. AB - The pontine parabrachial nucleus (PBN) has been implicated in regulating ingestion and contains opioids that promote feeding elsewhere in the brain. We tested the actions of the selective mu-opioid receptor (mu-OR) agonist [d-Ala2,N Me-Phe4,Gly5-ol]enkephalin (DAMGO) in the PBN on feeding in male rats with free access to food. Infusing DAMGO (0.5-4.0 nmol/0.5 microl) into the lateral parabrachial region (LPBN) increased food intake. The hyperphagic effect was anatomically specific to infusions within the LPBN, dose and time related, and selective for ingestion of chow compared with (nonnutritive) kaolin. The nonselective opioid antagonist naloxone (0.1-10.0 nmol intra-PBN) antagonized DAMGO-induced feeding, with complete blockade by 1.0 nmol and no effect on baseline. The highly selective mu-opioid antagonist d-Phe-Cys-Trp-Arg-Thr-Pen-Thr NH2 (CTAP; 1.0 nmol) also prevented this action of DAMGO, but the kappa antagonist nor-binaltorphimine did not. Naloxone and CTAP (10.0 nmol) decreased intake during scheduled feeding. Thus stimulating mu-ORs in the LPBN increases feeding, whereas antagonizing these sites inhibits feeding. Together, our results implicate mu-ORs in the LPBN in the normal regulation of food intake. PMID- 14557238 TI - Regulation of androgen receptor expression at the onset of functional overload in rat plantaris muscle. AB - Skeletal muscle androgen receptor (AR) expression at the onset of functional overload (OV) has not been well described. It is also not known if overload and/or anabolic steroid differentially regulate AR expression. The purpose of this study was to examine AR gene expression at the onset of functional OV in rat plantaris muscle with and without nandrolone decanoate (ND) administration. The functional significance of AR protein induction was examined using skeletal alpha actin promoter activity in transiently transfected CV-1 fibroblast cells. Male Sprague-Dawley rats ( approximately 125 g) were functionally overloaded for 1, 3, 7, or 21 days. A subset of animals was given an ND (6 mg/kg) injection at day 0 and then overloaded for 3 days. Control animals underwent sham surgeries. AR protein concentration increased 106 and 279% after 7 and 21 days of OV, respectively. AR mRNA increased 430% after 7 days of OV. AR protein expression in C2C12 murine myotubes subjected to 1% chronic radial stretch for 18 h was elevated 101% compared with control. ND treatment increased AR protein concentration 1,300% compared with controls, and there was no additional effect when ND and OV were combined. ND with 3 days of OV treatment increased AR mRNA expression 50% compared with control. AR overexpression in transiently transfected CV-1 fibroblast cells increased -424 bp skeletal alpha-actin promoter activity 80 to 1,800% in a dose-dependent fashion. Co-overexpression of either serum response factor (SRF) or active RhoA with AR overexpression induced a synergistic 36- and 28-fold induction of skeletal alpha-actin promoter. Cotransfection of AR, SRF, and active RhoA induced 180-fold increase in skeletal alpha-actin promoter activity. In conclusion, AR protein expression is increased after 7 days of functional OV, and this induction is regulated pretranslationally. AR induction in conjunction with SRF and RhoA signaling may be an important regulator of gene expression during overload-induced muscle growth. PMID- 14557239 TI - Increased thirst and vasopressin secretion after myocardial infarction in rats. AB - Impaired regulation of salt and water balance in left ventricular dysfunction and heart failure can lead to pulmonary and peripheral edema and hyponatremia. Previous studies of disordered water regulation in heart failure have used models of low cardiac output with normal cardiac function (e.g., inferior vena cava ligation). We investigated thirst and vasopressin (AVP) secretion in a rat myocardial infarction model of chronic left ventricular dysfunction/heart failure in response to a 24-h water deprivation period. Thirst (implied from water drunk), hematocrit, plasma renin activity, and plasma AVP concentrations increased with water deprivation vs. ad libitum water access. Thirst and plasma AVP concentrations were significantly positively correlated with infarct size after 24-h water deprivation but not under ad libitum water access conditions. The mechanism by which this occurs is unclear but could involve increased osmoreceptor sensitivity, altered stimulation of baroreceptors, the renin angiotensin system, or altered central neural control. PMID- 14557240 TI - C-reactive protein: a surrogate risk marker or mediator of atherothrombosis? PMID- 14557241 TI - C-reactive protein and atherothrombosis--beyond a biomarker: an actual partaker of lesion formation. PMID- 14557244 TI - Cdc48p is required for the cell cycle commitment point at Start via degradation of the G1-CDK inhibitor Far1p. AB - The budding yeast Cdc48p and its mammalian homologue p97 are involved in many important cellular activities. Because previous cdc48 mutants have exclusive G2/M arrest, Cdc48p was thought to play an essential role only during mitosis. We found that Cdc48p is required for the execution of Start (a yeast cell cycle commitment point equivalent to the restriction point in mammalian cells) in both a normal mitotic cell cycle and cell cycle reentry after mating pheromone withdrawal through degradation of the G1-cyclin-dependent kinase inhibitor Far1p. Our work is the first to uncover novel roles of Cdc48p as a critical cell cycle regulator in G1, and to shed new light on cell cycle regulation of Far1p, which is the first cyclin-dependent kinase inhibitor shown to be a substrate of an essential proteolysis event mediated by Cdc48p. PMID- 14557245 TI - APP-BP1 mediates APP-induced apoptosis and DNA synthesis and is increased in Alzheimer's disease brain. AB - APP-BP1, first identified as an amyloid precursor protein (APP) binding protein, is the regulatory subunit of the activating enzyme for the small ubiquitin-like protein NEDD8. We have shown that APP-BP1 drives the S- to M-phase transition in dividing cells, and causes apoptosis in neurons. We now demonstrate that APP-BP1 binds to the COOH-terminal 31 amino acids of APP (C31) and colocalizes with APP in a lipid-enriched fraction called lipid rafts. We show that coexpression of a peptide representing the domain of APP-BP1 that binds to APP, abolishes the ability of overexpressed APP or the V642I mutant of APP to cause neuronal apoptosis and DNA synthesis. A dominant negative mutant of the NEDD8 conjugating enzyme hUbc12, which participates in the ubiquitin-like pathway initiated by APP BP1, blocks neuronal apoptosis caused by APP, APP(V642I), C31, or overexpression of APP-BP1. Neurons overexpressing APP or APP(V642I) show increased APP-BP1 protein levels in lipid rafts. A similar increase in APP-BP1 in lipid rafts is observed in the Alzheimer's disease brain hippocampus, but not in less-affected areas of Alzheimer's disease brain. This translocation of APP-BP1 to lipid rafts is accompanied by a change in the subcellular localization of the ubiquitin-like protein NEDD8, which is activated by APP-BP1. PMID- 14557246 TI - AIP is a mitochondrial import mediator that binds to both import receptor Tom20 and preproteins. AB - Most mitochondrial preproteins are maintained in a loosely folded import competent conformation by cytosolic chaperones, and are imported into mitochondria by translocator complexes containing a preprotein receptor, termed translocase of the outer membrane of mitochondria (Tom) 20. Using two-hybrid screening, we identified arylhydrocarbon receptor-interacting protein (AIP), an FK506-binding protein homologue, interacting with Tom20. The extreme COOH terminal acidic segment of Tom20 was required for interaction with tetratricopeptide repeats of AIP. An in vitro import assay indicated that AIP prevents preornithine transcarbamylase from the loss of import competency. In cultured cells, overexpression of AIP enhanced preornithine transcarbamylase import, and depletion of AIP by RNA interference impaired the import. An in vitro binding assay revealed that AIP specifically binds to mitochondrial preproteins. Formation of a ternary complex of Tom20, AIP, and preprotein was observed. Hsc70 was also found to bind to AIP. An aggregation suppression assay indicated that AIP has a chaperone-like activity to prevent substrate proteins from aggregation. These results suggest that AIP functions as a cytosolic factor that mediates preprotein import into mitochondria. PMID- 14557247 TI - A role for Yip1p in COPII vesicle biogenesis. AB - Yeast Ypt1p-interacting protein (Yip1p) belongs to a conserved family of transmembrane proteins that interact with Rab GTPases. We encountered Yip1p as a constituent of ER-derived transport vesicles, leading us to hypothesize a direct role for this protein in transport through the early secretory pathway. Using a cell-free assay that recapitulates protein transport from the ER to the Golgi complex, we find that affinity-purified antibodies directed against the hydrophilic amino terminus of Yip1p potently inhibit transport. Surprisingly, inhibition is specific to the COPII-dependent budding stage. In support of this in vitro observation, strains bearing the temperature-sensitive yip1-4 allele accumulate ER membranes at a nonpermissive temperature, with no apparent accumulation of vesicle intermediates. Genetic interaction analyses of the yip1-4 mutation corroborate a function in ER budding. Finally, ordering experiments show that preincubation of ER membranes with COPII proteins decreases sensitivity to anti-Yip1p antibodies, indicating an early requirement for Yip1p in vesicle formation. We propose that Yip1p has a previously unappreciated role in COPII vesicle biogenesis. PMID- 14557248 TI - Identification of a novel antiapoptotic protein that antagonizes ASK1 and CAD activities. AB - Diverse stimuli initiate the activation of apoptotic signaling pathways that often causes nuclear DNA fragmentation. Here, we report a new antiapoptotic protein, a caspase-activated DNase (CAD) inhibitor that interacts with ASK1 (CIIA). CIIA, by binding to apoptosis signal-regulating kinase 1 (ASK1), inhibits oligomerization-induced ASK1 activation. CIIA also associates with CAD and inhibits the nuclease activity of CAD without affecting caspase-3-mediated ICAD cleavage. Overexpressed CIIA reduces H2O2- and tumor necrosis factor-alpha induced apoptosis. CIIA antisense oligonucleotides, which abolish expression of endogenous CIIA in murine L929 cells, block the inhibitory effect of CIIA on ASK1 activation, deoxyribonucleic acid fragmentation, and apoptosis. These findings suggest that CIIA is an endogenous antagonist of both ASK1- and CAD-mediated signaling. PMID- 14557249 TI - APP processing is regulated by cytoplasmic phosphorylation. AB - Amyloid-beta peptide (Abeta) aggregate in senile plaque is a key characteristic of Alzheimer's disease (AD). Here, we show that phosphorylation of amyloid precursor protein (APP) on threonine 668 (P-APP) may play a role in APP metabolism. In AD brains, P-APP accumulates in large vesicular structures in afflicted hippocampal pyramidal neurons that costain with antibodies against endosome markers and the beta-secretase, BACE1. Western blot analysis reveals increased levels of T668-phosphorylated APP COOH-terminal fragments in hippocampal lysates from many AD but not control subjects. Importantly, P-APP cofractionates with endosome markers and BACE1 in an iodixanol gradient and displays extensive colocalization with BACE1 in rat primary cortical neurons. Furthermore, APP COOH-terminal fragments generated by BACE1 are preferentially phosphorylated on T668 verses those produced by alpha-secretase. The production of Abeta is significantly reduced when phosphorylation of T668 is either abolished by mutation or inhibited by T668 kinase inhibitors. Together, these results suggest that T668 phosphorylation may facilitate the BACE1 cleavage of APP to increase Abeta generation. PMID- 14557250 TI - Huntingtin forms toxic NH2-terminal fragment complexes that are promoted by the age-dependent decrease in proteasome activity. AB - Although NH2-terminal mutant huntingtin (htt) fragments cause neurological disorders in Huntington's disease (HD), it is unclear how toxic htt fragments are generated and contribute to the disease process. Here, we report that complex NH2 terminal mutant htt fragments smaller than the first 508 amino acids were generated in htt-transfected cells and HD knockin mouse brains. These fragments constituted neuronal nuclear inclusions and appeared before neurological symptoms. The accumulation and aggregation of these htt fragments were associated with an age-dependent decrease in proteasome activity and were promoted by inhibition of proteasome activity. These results suggest that decreased proteasome activity contributes to late onset htt toxicity and that restoring the ability to remove NH2-terminal fragments will provide a more effective therapy for HD than inhibiting their production. PMID- 14557251 TI - Skeletal myosin heavy chain function in cultured lung myofibroblasts. AB - Myofibroblasts are unique contractile cells with both muscle and nonmuscle properties. Typically myofibroblasts are identified by the expression of alpha smooth muscle actin (ASMA); however some myofibroblasts also express sarcomeric proteins. In this study, we show that pulmonary myofibroblasts express three of the eight known sarcomeric myosin heavy chains (MyHCs) (IIa, IId, and embryonic) and that skeletal muscle myosin enzymatic activity is required for pulmonary myofibroblast contractility. Furthermore, inhibition of skeletal myosin activity and myofibroblast contraction results in a decrease in both ASMA and skeletal MyHC promoter activity and ASMA protein expression, suggesting a potential coupling of skeletal myosin activity and ASMA expression in myofibroblast differentiation. To understand the molecular mechanisms whereby skeletal muscle genes are regulated in myofibroblasts, we have found that members of the myogenic regulatory factor family of transcription factors and Ca(2+) - regulated pathways are involved in skeletal MyHC promoter activity. Interestingly, the regulation of skeletal myosin expression in myofibroblasts is distinct from that observed in muscle cells and suggests that cell context is important in its control. PMID- 14557252 TI - How VASP enhances actin-based motility. AB - The function of vasodilator-stimulated phosphoprotein (VASP) in motility is analyzed using a biomimetic motility assay in which ActA-coated microspheres propel themselves in a medium containing actin, the Arp2/3 complex, and three regulatory proteins in the absence or presence of VASP. Propulsion is linked to cycles of filament barbed end attachment-branching-detachment-growth in which the ActA-activated Arp2/3 complex incorporates at the junctions of branched filaments. VASP increases the velocity of beads. VASP increases branch spacing of filaments in the actin tail, as it does in lamellipodia in living cells. The effect of VASP on branch spacing of Arp2/3-induced branched actin arrays is opposed to the effect of capping proteins. However, VASP does not compete with capping proteins for binding barbed ends of actin filaments. VASP enhances branched actin polymerization only when ActA is immobilized on beads or on Listeria. VASP increases the rate of dissociation of the branch junction from immobilized ActA, which is the rate-limiting step in the catalytic cycle of site directed filament branching. PMID- 14557253 TI - CD151 regulates epithelial cell-cell adhesion through PKC- and Cdc42-dependent actin cytoskeletal reorganization. AB - CD151, a member of the tetraspanin family proteins, tightly associates with integrin alpha3beta1 and localizes at basolateral surfaces of epithelial cells. We found that overexpression of CD151 in A431 cells accelerated intercellular adhesion, whereas treatment of cells with anti-CD151 mAb perturbed the integrity of cortical actin filaments and cell polarity. E-Cadherin puncta formation, indicative of filopodia-based adhesion zipper formation, as well as E-cadherin anchorage to detergent-insoluble cytoskeletal matrix, was enhanced in CD151 overexpressing cells. Levels of GTP-bound Cdc42 and Rac were also elevated in CD151-overexpressing cells, suggesting the role of CD151 in E-cadherin-mediated cell-cell adhesion as a modulator of actin cytoskeletal reorganization. Consistent with this possibility, engagement of CD151 by the substrate-adsorbed anti-CD151 mAb induced prominent Cdc42-dependent filopodial extension, which along with E-cadherin puncta formation, was strongly inhibited by calphostin C, a protein kinase C (PKC) inhibitor. Together, these results indicate that CD151 is involved in epithelial cell-cell adhesion as a modulator of PKC- and Cdc42 dependent actin cytoskeletal reorganization. PMID- 14557254 TI - Distinct ligand binding sites in integrin alpha3beta1 regulate matrix adhesion and cell-cell contact. AB - The integrin alpha3beta1 mediates cellular adhesion to the matrix ligand laminin 5. A second integrin ligand, the urokinase receptor (uPAR), associates with alpha3beta1 via a surface loop within the alpha3 beta-propeller (residues 242 246) but outside the laminin binding region, suggesting that uPAR-integrin interactions could signal differently from matrix engagement. To explore this, alpha3-/- epithelial cells were reconstituted with wild-type (wt) alpha3 or alpha3 with Ala mutations within the uPAR-interacting loop (H245A or R244A). Wt or mutant-bearing cells showed comparable expression and adhesion to laminin-5. Cells expressing wt alpha3 and uPAR dissociated in culture, with increased Src activity, up-regulation of SLUG, and down-regulation of E-cadherin and gamma catenin. Src kinase inhibition or expression of Src 1-251 restored the epithelial phenotype. The H245A and R244A mutants were unaffected by coexpression of uPAR. We conclude that alpha3beta1 regulates both cell-cell contact and matrix adhesion, but through distinct protein interaction sites within its beta propeller. These studies reveal an integrin- and Src-dependent pathway for SLUG expression and mesenchymal transition. PMID- 14557255 TI - Direct determination of the interleukin-6 binding epitope of the interleukin-6 receptor by NMR spectroscopy. AB - All cytokines belonging to the interleukin-6 (IL-6)-type family of cytokines utilize receptors that have a modular build of several immunoglobulin-like and fibronectin type III-like domains. Characteristic of these receptors is a cytokine receptor homology region consisting of two such fibronectin domains defined by a set of four conserved cysteines and a tryptophan-serine-X-tryptophan serine sequence motif. On target cells, interleukin-6 first binds to its specific receptor and subsequently to a homodimer of the signal transducer protein gp130. The interleukin-6 receptor consists of three extracellular domains. The N terminal immunoglobulin-like domain is not involved in ligand binding, whereas the third membrane proximal fibronectin-like domain accounts for more than 90% of the binding energy to IL-6. Here, the key residues of this fibronectin-like domain involved in the interaction with IL-6 are described. Chemical shift mapping data with 15N-labeled IL-6R-D3 and unlabeled IL-6 coupled with recent structural data clearly reveal the epitope within the IL-6R-D3 responsible for mediating the high affinity interaction with its cognate cytokine. PMID- 14557256 TI - Tumor necrosis factor receptor-associated factor (TRAF) 1 regulates CD40-induced TRAF2-mediated NF-kappaB activation. AB - To investigate CD40 signaling complex formation in living cells, we used green fluorescent protein (GFP)-tagged CD40 signaling intermediates and confocal life imaging. The majority of cytoplasmic TRAF2-GFP and, to a lesser extent, TRAF3 GFP, but not TRAF1-GFP or TRAF4-GFP, translocated into CD40 signaling complexes within a few minutes after CD40 triggering with the CD40 ligand. The inhibitor of apoptosis proteins cIAP1 and cIAP2 were also recruited by TRAF2 to sites of CD40 signaling. An excess of TRAF2 allowed recruitment of TRAF1-GFP to sites of CD40 signaling, whereas an excess of TRAF1 abrogated the interaction of TRAF2 and CD40. Overexpression of TRAF1, however, had no effect on the interaction of TRADD and TRAF2, known to be important for tumor necrosis factor receptor 1 (TNF-R1) mediated NF-kappaB activation. Accordingly, TRAF1 inhibited CD40-dependent but not TNF-R1-dependent NF-kappaB activation. Moreover, down-regulation of TRAF1 with small interfering RNAs enhanced CD40/CD40 ligand-induced NF-kappaB activation but showed no effect on TNF signaling. Because of the trimeric organization of TRAF proteins, we propose that the stoichiometry of TRAF1-TRAF2 heteromeric complexes ((TRAF2)2-TRAF1 versus TRAF2-(TRAF1)2) determines their capability to mediate CD40 signaling but has no major effect on TNF signaling. PMID- 14557257 TI - Gene fusion and overlapping reading frames in the mammalian genes for 4E-BP3 and MASK. AB - 4E-BP3 is a member of the eukaryotic initiation factor (eIF) 4F-binding protein family of translational repressors. eIF4E-binding proteins (4E-BPs) inhibit translation initiation by sequestering eIF4E, the cap-binding protein, from eIF4G thus preventing ribosome recruitment to the mRNA. Previous analysis of 4E-BP3 expression uncovered an 8.5-kb mRNA variant of unknown origin. To study this splice variant, we determined the structure of the genomic locus encoding human 4E-BP3 (EIF4EBP3). EIF4EBP3 is located on human chromosome 5q31.3 and comprises three exons (A, B, and C) and two introns. Exon B contains the region of the open reading frame responsible for eIF4E binding. GenBank searches revealed multiple expressed sequence tags originating from the alternative splicing of exon B with unidentified upstream exons. Further studies revealed that the 8.5-kb transcript arises from the fusion of EIF4EBP3 with the mammalian homologue of Drosophila MASK (multiple ankyrin repeats, single KH domain), which is crucial for photoreceptor differentiation, cell survival, and proliferation. Surprisingly, the open reading frame of the MASK-BP3 transcript is different from that of 4E BP3, which indicates that exon B is translated using an alternative reading frame. A gene fusion similar to that of MASK and EIF4EBP3 has been reported only once in mammals for the UEV1-Kua transcript. The use of an alternative reading frame is also very rare, having been described for two loci, INK4a/ARF and XLalphas/ALEX. The simultaneous exploitation of both mechanisms underscores the flexibility of mammalian genomes and has important implications for the functional analysis of 4E-BP3 and MASK. Interestingly, both eIF4E and MASK are downstream effectors of the Ras/MAPK pathway, which provides a rationale for the MASK-BP3 fusion in mammals. PMID- 14557258 TI - Lysine-phosphatidylcholine adducts in kringle V impart unique immunological and potential pro-inflammatory properties to human apolipoprotein(a). AB - Lipoprotein(a), Lp(a), an athero-thrombotic risk factor, reacts with EO6, a natural monoclonal autoantibody that recognizes the phophorylcholine (PC) group of oxidized phosphatidylcholine (oxPtdPC) either as a lipid or linked by a Schiff base to lysine residues of peptides/proteins. Here we show that EO6 reacts with free apolipoprotein(a) apo(a), its C-terminal domain, F2 (but not the N-terminal F1), kringle V-containing fragments obtained by the enzymatic digestion of apo(a) and also kringle V-containing apo(a) recombinants. The evidence that kringle V is critical for EO6 reactivity is supported by the finding that apo(a) of rhesus monkeys lacking kringle V did not react with EO6. Based on the previously established EO6 specificity requirements, we hypothesized that all or some of the six lysines in human kringle V are involved in Schiff base linkage with oxPtdPC. To test this hypothesis, we made use of a recombinant lysine-containing apo(a) fragment, rIII, containing kringle V but not the protease domain. EO6 reacted with rIII before and after reduction to stabilize the Schiff base and also after extensive ethanol/ether extraction that yielded no lipids. On the other hand, delipidation of the saponified product yielded an average of two mol of phospholipids/mol of protein consistent with direct analysis of inorganic phosphorous on the non-saponified rIII. Moreover, only two of the six theoretical free lysine amino groups per mol of rIII were unavailable to chemical modification by 2,4,6-trinitrobenzene sulfonic acid. Finally, rIII, like human apo(a), stimulated the production of interleukin 8 in THP-1 macrophages in culture. Together, our studies provide evidence that in human apo(a), kringle V is the site that reacts with EO6 via lysine-oxPtdPC adducts that may also be involved in the previously reported pro-inflammatory effect of apo(a) in cultured human macrophages. PMID- 14557260 TI - Mutagenesis of putative catalytic and regulatory residues of Streptomyces chromofuscus phospholipase D differentially modifies phosphatase and phosphodiesterase activities. AB - Phospholipase D from Streptomyces chromofuscus (sc-PLD) is a member of the diverse family of metallo-phosphodiesterase/phosphatase enzymes that also includes purple acid phosphatases, protein phosphatases, and nucleotide phosphodiesterases. Whereas iron is an essential cofactor for scPLD activity, Mn2+ is also found in the enzyme. A third metal ion, Ca2+, has been shown to enhance scPLD catalytic activity although it is not an essential cofactor. Sequence alignment of scPLD with known phosphodiesterases and phosphatases requiring metal ions suggested that His-212, Glu-213, and Asp-389 could be involved in Mn2+ binding. H212A, E213A, and D389A were prepared to test this hypothesis. These three mutant enzymes and wild type scPLD show similar metal content but considerably different catalytic properties, suggesting different roles for each residue. His-212 appears involved in binding the phosphate group of substrates, whereas Glu-213 acts as a ligand for Ca2+. D389A showed a greatly reduced phosphodiesterase activity but almost unaltered ability to hydrolyze the phosphate group in p-nitrophenyl phosphate suggesting it had a critical role in aligning groups at the active site to control phosphodiesterase versus phosphatase activities. We propose a model for substrate and cofactor binding to the catalytic site of scPLD based on these results and on sequence alignment to purple acid phosphatases of known structure. PMID- 14557259 TI - Adiponectin stimulates angiogenesis by promoting cross-talk between AMP-activated protein kinase and Akt signaling in endothelial cells. AB - Adiponectin is an adipocyte-specific adipocytokine with anti-atherogenic and anti diabetic properties. Here, we investigated whether adiponectin regulates angiogenic processes in vitro and in vivo. Adiponectin stimulated the differentiation of human umbilical vein endothelium cells (HUVECs) into capillary like structures in vitro and functioned as a chemoattractant in migration assays. Adiponectin promoted the phosphorylation of AMP-activated protein kinase (AMPK), protein kinase Akt/protein kinase B, and endothelial nitric oxide synthesis (eNOS) in HUVECs. Transduction with either dominant-negative AMPK or dominant negative Akt abolished adiponectin-induced eNOS phosphorylation as well as adiponectin-stimulated HUVEC migration and differentiation. Dominant-negative AMPK also inhibited adiponectin-induced Akt phosphorylation, suggesting that AMPK is upstream of Akt. Dominant-negative Akt or the phosphatidylinositol 3-kinase inhibitor LY294002 blocked adiponectin-stimulated Akt and eNOS phosphorylation, migration, and differentiation without altering AMPK phosphorylation. Finally, adiponectin stimulated blood vessel growth in vivo in mouse Matrigel plug implantation and rabbit corneal models of angiogenesis. These data indicate that adiponectin can function to stimulate the new blood vessel growth by promoting cross-talk between AMP-activated protein kinase and Akt signaling within endothelial cells. PMID- 14557261 TI - The three-dimensional structure of the N-acetylglucosamine-6-phosphate deacetylase, NagA, from Bacillus subtilis: a member of the urease superfamily. AB - The enzyme N-acetylglucosamine-6-phosphate deacetylase, NagA, catalyzes the hydrolysis of the N-acetyl group of GlcNAc-6-P to yield glucosamine 6-phosphate and acetate, the first committed step in the biosynthetic pathway to amino-sugar nucleotides. It is classified into carbohydrate esterase family CE-9 (see afmb.cnrs-mrs.fr/CAZY/). Here we report the cloning, expression, and three dimensional structure (Protein Data Bank code 1un7) determination by x-ray crystallography of the Bacillus subtilis NagA at a resolution of 2.0 A. The structure presents two domains, a (beta/alpha)(8) barrel enclosing the active center and a small beta barrel domain. The structure is dimeric, and the substrate phosphate coordination at the active center is provided by an Arg/His pair contributed from the second molecule of the dimer. Both the overall structure and the active center bear a striking similarity to the urease superfamily with two metals involved in substrate binding and catalysis. PIXE (Proton-Induced x-ray Emission) data show that iron is the predominant metal in the purified protein. We propose a catalytic mechanism involving proton donation to the leaving group by aspartate, nucleophilic attack by an Fe-bridged hydroxide, and stabilization of the carbonyl oxygen by one of the two Fe atoms of the pair. We believe that this is the first sugar deacetylase to utilize this fold and catalytic mechanism. PMID- 14557262 TI - Contribution of the membrane-distal tyrosine in intracellular signaling by the granulocyte colony-stimulating factor receptor. AB - We have evaluated the contribution of intracellular tyrosine residues of the granulocyte colony-stimulating factor receptor (GCSF-R) to its signaling and cellular outcomes. We began with stable BaF3 cell lines overexpressing wild-type or mutant GCSF-Rs. When all four intracellular tyrosines of the GCSF-R were replaced with phenylalanine (FFFF GCSF-R), cell proliferation and survival were compromised. Replacement of only the membrane-distal tyrosine (YYYF GCSF-R) also showed reduced survival following a GCSF withdrawal/replacement protocol, suggesting a role for this tyrosine. Proliferation by FFFY GCSF-R cells was attenuated by approximately 70%. In evaluating the biochemical steps involved in signaling, we then showed that the membrane-distal tyrosine was necessary and sufficient for c-Jun N-terminal kinase (JNK) activation. With the use of a cell permeable JNK-inhibitory peptide, JNK was implicated in the proliferation of the FFFY GCSF-R mutant. To further define the events linking the membrane-distal tyrosine and JNK activation, the Src homology 2 domains of Shc, Grb2, and 3BP2 were shown to bind the full-length GCSF-R and a phosphopeptide encompassing the membrane-distal tyrosine. When binding to variant phosphopeptides based on this membrane-distal tyrosine was tested, altering the amino acids immediately following the phosphotyrosine could selectively abolish the interaction with Shc or Grb2, or the binding to both Grb2 and 3BP2. When these changes were introduced into the full-length GCSF-R and new cell lines created, only the mutant that did not interact with Grb2 and 3BP2 did not activate JNK. Our results suggest that direct binding of Shc by the GCSF-R is not essential for JNK activation. PMID- 14557263 TI - Phosphorylation-regulated inhibition of the Gz GTPase-activating protein activity of RGS proteins by synapsin I. AB - Synapsins are neuronal proteins that bind and cluster synaptic vesicles in the presynaptic space, presumably by anchoring to actin filaments, but specific regulatory functions of the synapsins are unknown. We found that a sub-population of brain synapsin Ia, a splice variant of one of three synapsin isoforms, inhibits the GTPase-activating protein (GAP) activity of several RGS proteins. Inhibition is highly selective for Galphaz, a member of the Gi family that is found in neurons, platelets, adrenal chromaffin cells, and a few other neurosecretory cells. Gz has been indirectly implicated in the regulation of secretion. Synapsin Ia constitutes a major fraction of the total GAP-inhibitory activity in brain, and its inhibitory activity is absent from the brains of synapsin I(-/-)/II(-/-) mice. Inhibition depends on the cationic D/E domain of synapsin. Phosphorylation of synapsin Ia at serine 9 by either cyclic AMP dependent protein kinase or p21-activated protein kinase (PAK1) attenuates its potency as a GAP inhibitor more than 7-fold. Synapsin can thus act as a phosphorylation-modulated mediator of feedback regulation of Gz signaling by the synaptic machinery. PMID- 14557264 TI - Mannose 6-phosphate receptor-mediated uptake is defective in acid sphingomyelinase-deficient macrophages: implications for Niemann-Pick disease enzyme replacement therapy. AB - Progressive accumulation of lipid-laden macrophages is a hallmark of the acid sphingomyelinase (ASM)-deficient forms of Niemann-Pick disease (i.e. Types A and B NPD). To investigate the mechanisms underlying enzyme replacement therapy for this disorder, we studied the uptake of recombinant, human ASM (rhASM) by alveolar macrophages from ASM knock-out (ASMKO) mice. The recombinant enzyme used for these studies was produced in Chinese hamster ovary cells and contained complex type, N-linked oligosaccharides. Binding of radiolabeled, rhASM to the ASMKO macrophages was enhanced as compared with normal macrophages, consistent with their larger size and increased surface area. However, internalization of the enzyme by the ASMKO cells was markedly reduced when compared with normal cells. Studies using receptor-specific ligands to inhibit enzyme uptake revealed that in normal cells rhASM was taken up by a combination of mannose and mannose 6 phosphate receptors (MR and M6PR, respectively), whereas in the ASMKO cells the M6PR had a minimal role in rhASM uptake. Expression of M6PR mRNA was normal in the ASMKO cells, although Western blotting revealed more receptors in these cells when compared with normal. We therefore hypothesized that lipid accumulation in ASMKO macrophages led to abnormalities in M6PR trafficking and/or degradation, resulting in reduced enzyme uptake. Consistent with this hypothesis, we also found that, when rhASM was modified to expose terminal mannose residues and target mannose receptors, the uptake of this modified enzyme form by ASMKO cells was approximately 10-fold greater when compared with the "complex" type rhASM. These findings have important implications for NPD enzyme replacement therapy, particularly in the lung. PMID- 14557265 TI - Two components of a secreted cell number-counting factor bind to cells and have opposing effects on cAMP signal transduction in Dictyostelium. AB - A secreted 450-kDa complex of proteins called counting factor (CF) is part of a negative feedback loop that regulates the size of the groups formed by developing Dictyostelium cells. Two components of CF are countin and CF50. Both recombinant countin and recombinant CF50 decrease group size in Dictyostelium. countin- cells have a decreased cAMP-stimulated cAMP pulse, whereas recombinant countin potentiates the cAMP pulse. We find that CF50 cells have an increased cAMP pulse, whereas recombinant CF50 decreases the cAMP pulse, suggesting that countin and CF50 have opposite effects on cAMP signal transduction. In addition, countin and CF50 have opposite effects on cAMP-stimulated Erk2 activation. However, like recombinant countin, recombinant CF50 increases cell motility. We previously found that cells bind recombinant countin with a Hill coefficient of approximately 2, a KH of 60 pm, and approximately 53 sites/cell. We find here that cells also bind 125I-recombinant CF50, with a Hill coefficient of approximately 2, a KH of approximately 15 ng/ml (490 pm), and approximately 56 sites/cell. Countin and CF50 require each other's presence to affect group size, but the presence of countin is not necessary for CF50 to bind to cells, and CF50 is not necessary for countin to bind to cells. Our working hypothesis is that a signal transduction pathway activated by countin binding to cells modulates a signal transduction pathway activated by CF50 binding to cells and vice versa and that these two pathways can be distinguished by their effects on cAMP signal transduction. PMID- 14557266 TI - Mechanism and stoichiometry of interaction of DnaG primase with DnaB helicase of Escherichia coli in RNA primer synthesis. AB - Initiation and synthesis of RNA primers in the lagging strand of the replication fork in Escherichia coli requires the replicative DnaB helicase and the DNA primase, the DnaG gene product. In addition, the physical interaction between these two replication enzymes appears to play a role in the initiation of chromosomal DNA replication. In vitro, DnaB helicase stimulates primase to synthesize primers on single-stranded (ss) oligonucleotide templates. Earlier studies hypothesized that multiple primase molecules interact with each DnaB hexamer and single-stranded DNA. We have examined this hypothesis and determined the exact stoichiometry of primase to DnaB hexamer. We have also demonstrated that ssDNA binding activity of the DnaB helicase is necessary for directing the primase to the initiator trinucleotide and synthesis of 11-20-nucleotide long primers. Although, association of these two enzymes determines the extent and rate of synthesis of the RNA primers in vitro, direct evidence of the formation of primase-DnaB complex has remained elusive in E. coli due to the transient nature of their interaction. Therefore, we stabilized this complex using a chemical cross-linker and carried out a stoichiometric analysis of this complex by gel filtration. This allowed us to demonstrate that the primase-helicase complex of E. coli is comprised of three molecules of primase bound to one DnaB hexamer. Fluorescence anisotropy studies of the interaction of DnaB with primase, labeled with the fluorescent probe Ru(bipy)3, and Scatchard analysis further supported this conclusion. The addition of DnaC protein, leading to the formation of the DnaB-DnaC complex, to the simple priming system resulted in the synthesis of shorter primers. Therefore, interactions of the DnaB-primase complex with other replication factors might be critical for determining the physiological length of the RNA primers in vivo and the overall kinetics of primer synthesis. PMID- 14557267 TI - Interferon regulatory factor-3-mediated activation of the interferon-sensitive response element by Toll-like receptor (TLR) 4 but not TLR3 requires the p65 subunit of NF-kappa. AB - Interferon regulatory factor (IRF) 3 is a transcription factor that binds the interferon-sensitive response element (ISRE) and is activated by Toll-like receptor 3 (TLR3) and TLR4. We have found that a dominant negative form of I kappa B kinase 2 and a mutant form of I kappa B, which acts as a super-repressor of NF-kappa B, blocked activation of the ISRE by the TLR4 ligand lipopolysaccharide but not the TLR3 ligand poly(I-C). TLR4 failed to activate the ISRE in mouse embryonic fibroblasts bearing a targeted deletion of p65, whereas the response to TLR3 in these cells was normal. The p65 subunit of NF-kappa B was detected in the lipopolysaccharide-activated but not poly(I-C)-activated ISRE binding complex. Finally, p65 promoted transactivation of gene expression by IRF 3. These results therefore indicate that IRF-3-mediated activation of the ISRE by TLR4 but not TLR3 requires the p65 subunit of NF-kappa B. PMID- 14557268 TI - G protein-coupled receptor kinase interaction with Hsp90 mediates kinase maturation. AB - G protein-coupled receptor kinase 2 (GRK2) is a serine/threonine-specific protein kinase that mediates agonist-dependent phosphorylation of numerous G protein coupled receptors. In an effort to identify proteins that regulate GRK2 function, we searched for interacting proteins by immunoprecipitation of endogenous GRK2 from HL60 cells. Subsequent analysis by gel electrophoresis and mass spectrometry revealed that GRK2 associates with heat shock protein 90 (Hsp90). GRK2 interaction with Hsp90 was confirmed by co-immunoprecipitation and was effectively disrupted by geldanamycin, an Hsp90-specific inhibitor. Interestingly, geldanamycin treatment of HL60 cells decreased the expression of endogenous GRK2 in a dose- and time-dependent manner, and metabolic labeling demonstrated that geldanamycin rapidly accelerated the degradation of newly synthesized GRK2. The use of various protease inhibitors suggested that GRK2 degradation induced by geldanamycin was predominantly through the proteasome pathway. To test whether Hsp90 plays a general role in regulating GRK maturation, additional GRKs were studied by transient expression in COS-1 cells and subsequent treatment with geldanamycin. These studies demonstrate that GRK3, GRK5, and GRK6 are also stabilized by interaction with Hsp90. Taken together, our work revealed that GRK interaction with heat shock proteins plays an important role in regulating GRK maturation. PMID- 14557269 TI - Up-regulation of cyclooxygenase-2 and apoptosis resistance by p38 MAPK in hypericin-mediated photodynamic therapy of human cancer cells. AB - Photodynamic Therapy (PDT) is an approved anticancer therapy that kills cancer cells by the photochemical generation of reactive oxygen species following absorption of visible light by a photosensitizer, which selectively accumulates in tumors. We report that hypericin-mediated PDT of human cancer cells leads to up-regulation of the inducible cyclooxygenase-2 (COX-2) enzyme and the subsequent release of PGE2. Dissection of the signaling pathways involved revealed that the selective activation of p38 MAPK alpha and beta mediate COX-2 up-regulation at the protein and messenger levels. The p38 MAPK inhibitor, PD169316, abrogated COX 2 expression in PDT-treated cells, whereas overexpression of the drug-resistant PD169316-insensitive p38 MAPK alpha and beta isoforms restored COX-2 levels in the presence of the kinase inhibitor. Transcriptional regulation by nuclear factor-kappaB was not involved in COX-2 up-regulation by PDT. The half-life of the COX-2 messenger was drastically shortened by p38 MAPK inhibition in transcriptionally arrested cells, suggesting that p38 MAPK mainly acts by stabilizing the COX-2 transcript. Overexpression of WT-p38 MAPK increased cellular resistance to PDT-induced apoptosis, and inhibiting this pathway exacerbated cell death and prevented PGE2 secretion. Hence, the combination of PDT with pyridinyl imidazole inhibitors of p38 MAPK may improve the therapeutic efficacy of PDT by blocking COX-2 up-regulation, which contributes to tumor growth by the release of growth- and pro-angiogenic factors, as well as by sensitizing cancer cells to apoptosis. PMID- 14557270 TI - Basic fibroblast growth factor stimulates activation of Rac1 through a p85 betaPIX phosphorylation-dependent pathway. AB - In a previous study (Shin, E. Y., Shin, K. S., Lee, C. S., Woo, K. N., Quan, S. H., Soung, N. K., Kim, Y. G., Cha, C. I., Kim, S. R., Park, D., Bokoch, G. M., and Kim, E. G. (2002) J. Biol. Chem. 277, 44417-44430) we reported that phosphorylation of p85 betaPIX, a guanine nucleotide exchange factor (GEF) for Rac1/Cdc42, is a signal for translocation of the PIX complex to neuronal growth cones and is associated with basic fibroblast growth factor (bFGF)-induced neurite outgrowth. However, the issue of whether p85 betaPIX phosphorylation affects GEF activity on Rac1/Cdc42 is yet to be explored. Here we show that Rac1 activation occurs in a p85 betaPIX phosphorylation-dependent manner. A GST-PBD binding assay reveals that Rac1 is activated in a dose- and time-dependent manner in PC12 cells in response to bFGF. Inhibition of ERK or PAK2, the kinases upstream of p85 betaPIX in the bFGF signaling, prevents Rac1 activation, suggesting that phosphorylation of p85 betaPIX functions upstream of Rac1 activation. To directly address this issue, transfection studies with wild-type and mutant p85 betaPIX (S525A/T526A, a non-phosphorylatable form) were performed. Expression of mutant PIX markedly inhibits both bFGF- and nerve growth factor (NGF)-induced activation of Rac1, indicating that phosphorylation of p85 betaPIX is responsible for activation of this G protein. Both wild-type and mutant p85 betaPIX displaying negative GEF activity (L238R/L239S) are similarly recruited to growth cones, suggesting that Rac1 activation is not essential for translocation of the PIX complex (PAK2-p85 betaPIX-Rac1). However, expression of mutant p85 betaPIX (L238R/L239S) results in retraction of the pre-existing neurites. Our results provide evidence that bFGF- and NGF-induced phosphorylation of p85 betaPIX mediates Rac1 activation, which in turn regulates cytoskeletal reorganization at growth cones, but not translocation of the PIX complex. PMID- 14557271 TI - Characterization of a low molecular weight glycolipid antigen from Cryptosporidium parvum. AB - Cryptosporidium parvum, an Apicomplexan parasite of the mammalian gut epithelium, causes a diarrheal illness in a wide range of hosts and is transmitted by contamination of food or water with oocyst-laden feces from an infected animal. We have identified a glycosylinositol phospholipid from the sporozoite stage of the parasite that is frequently recognized by serum antibodies from human cryptosporidiosis patients. The humoral immune response is dominated by IgG1 subclass antibodies but can also include IgA and IgM antibodies. The glycosylinositol phospholipids were purified by butanol extraction of a Triton X 114-soluble fraction followed by octyl-Sepharose column chromatography and preparative high performance TLC and were shown to include at least 5 species. By using mass spectrometry and radiolabeled neutral glycan analysis, we found that the structure of the dominant glycosylinositol phospholipid antigen contained a C18:0 lyso-acylglycerol, a C16:0-acylated inositol, and an unsubstituted mannose3 glucosamine glycan core. Other diacyl species were also identified, most notably a series of glycosylinositol phospholipids having an acyl-linked C20:0 to C28:0 lipid on the inositol ring. Less abundant species having three acyl-linked fatty acids and species with an additional 1-3 hexoses linked to the mannose core were also observed. We are currently working to determine the role that these glycolipids may play in the development of disease and in the clearance of infection. PMID- 14557272 TI - Residue Gln4863 within a predicted transmembrane sequence of the Ca2+ release channel (ryanodine receptor) is critical for ryanodine interaction. AB - Despite the pivotal role of ryanodine in ryanodine receptor (RyR) research, the molecular basis of ryanodine-RyR interaction remains largely undefined. We investigated the role of the proposed transmembrane helix TM10 in ryanodine interaction and channel function. Each amino acid residue within the TM10 sequence, 4844IIFDITFFFFVIVILLAIIQGLII4867, of the mouse RyR2 was mutated to either alanine or glycine. Mutants were expressed in human embryonic kidney 293 cells, and their properties were assessed. Mutations D4847A, F4850A, F4851A, L4858A, L4859A, and I4866A severely curtailed the release of intracellular Ca2+ in human embryonic kidney 293 cells in response to extracellular caffeine and diminished [3H]ryanodine binding to cell lysates. Mutations F4846A, T4849A, I4855A, V4856A, and Q4863A eliminated or markedly reduced [3H]ryanodine binding, but cells expressing these mutants responded to extracellular caffeine by releasing stored Ca2+. Interestingly these two groups of mutants, each with similar properties, are largely located on opposite sides of the predicted TM10 helix. Single channel analyses revealed that mutation Q4863A dramatically altered the kinetics and apparent affinity of ryanodine interaction with single RyR2 channels and abolished the effect of ryanodol, an analogue of ryanodine, whereas the single channel conductance of the Q4863A mutant and its responses to caffeine, ATP, and Mg2+ were comparable to those of the wild type channels. Furthermore the effect of ryanodine on single Q4863A mutant channels was influenced by the transmembrane holding potential. Together these results suggest that the TM10 sequence and in particular the Q4863 residue constitute an important determinant of ryanodine interaction. PMID- 14557273 TI - 15-Deoxy-Delta12,14-prostaglandin J2 regulates endogenous Cot MAPK kinase kinase 1 activity induced by lipopolysaccharide. AB - Cot is a MAPK kinase kinase that has been implicated in cellular activation and proliferation. Here, we show that the addition of lipopolysaccharide (LPS) to RAW264 macrophages induces a 10-fold increase of endogenous Cot activity, measured as MAPK kinase kinase 1 activity. Taxol, but not phorbol 12-myristate 13 acetate (PMA), induces a similar activation of Cot. A tyrosine kinase activity is involved in Cot activation by LPS. 15-Deoxy-Delta12,14-prostaglandin J2, but not rosiglitazone, blocks Cot activation by LPS. Furthermore, 15-deoxy-Delta12,14 prostaglandin J2 also inhibited the LPS-induced Cot in vitro. However, 15-deoxy Delta12,14-prostaglandin J2 does not inhibit MAPK kinase 1 or ERK1/ERK2 activation/phosphorylation induced by PMA and mediated by c-Raf. Considering these data, we propose that the inhibition of LPS-induced Cot activation is one mechanism by which 15-deoxy-Delta12,14-prostaglandin J2 acts as an anti inflammatory. PMID- 14557274 TI - Differential and special properties of the major human UGT1-encoded gastrointestinal UDP-glucuronosyltransferases enhance potential to control chemical uptake. AB - UDP-glucuronosyltransferase (UGT) isozymes detoxify metabolites, drugs, toxins, and environmental chemicals via conjugation to glucuronic acid. Based on the extended UGT1 locus combined with Northern blot analysis and in situ hybridization, we determined the distribution of UGT1A1 and UGT1A7 through UGT1A10 mRNAs and found them for the first time segmentally distributed in the mucosal epithelia layer of the gastrointestinal tract. Biochemically, recombinant isozymes exhibited pH optima of 5.5, 6.4, 7.6, 8.5, and/or a broad pH range, and activities were found to be unaffected or progressively inhibited by increasing substrate concentrations after attaining Vmax for certain chemicals. Under different optimal conditions, all exhibited wide substrate selections for dietary and environmentally associated chemicals. Evidence also suggests tandem effects of isozymes in the time for completion of reactions when comparing short- and long-term incubations. Moreover, treatment of colon cells with certain diet associated constituents, curcumin and nordihydroguaiaretic acid, reversibly targets UGTs causing inhibition without affecting protein levels; there is no direct inhibition of control UGT using curcumin as substrate in the in vitro assay. In summary, we demonstrate that UGTs are located in gastrointestinal mucosa, have vast overlapping activities under differential optimal conditions, and exhibit marked sensitivity to certain dietary substrates/constituents, representing a first comprehensive study of critical properties concerning glucuronidating isozymes in alimentary tissues. Additionally, the highly dynamic, complex, and variable properties necessarily impact absorption of ingested chemicals and therapeutic drugs. PMID- 14557275 TI - Overexpression of diacylglycerol acyltransferase-1 reduces phospholipid synthesis, proliferation, and invasiveness in simian virus 40-transformed human lung fibroblasts. AB - Diacylglycerol (DAG) is a versatile molecule that participates as substrate in the synthesis of structural and energetic lipids, and acts as the physiological signal that activates protein kinase C. Diacylglycerol acyltransferase (DGAT), the last committed enzyme in triacylglycerol synthesis, could potentially regulate the content and use of both signaling and glycerolipid substrate DAG by converting it into triacylglycerol. To test this hypothesis, we stably overexpressed the DGAT1 mouse gene in human lung SV40-transformed fibroblasts (DGAT cells), which contains high levels of DAG. DGAT cells exhibited a 3.9-fold higher DGAT activity and a 3.2-fold increase in triacylglycerol content, whereas DAG and phosphatidylcholine decreased by 70 and 20%, respectively, compared with empty vector-transfected SV40 cells (Control cells). Both acylation and de novo synthesis of phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin were reduced by 30-40% in DGAT cells compared with controls, suggesting that DGAT used substrates for triacylglycerol synthesis that had originally been destined to produce phospholipids. The incorporation of [14C]DAG and [14C]fatty acids released from plasma membrane by additions of either phospholipase C or phospholipase A2 into triacylglycerol was increased by 6.2- and 2.8-fold, respectively, in DGAT cells compared with control cells, indicating that DGAT can attenuate signaling lipids. Finally, DGAT overexpression reversed the neoplastic phenotype because it dramatically reduced the cell growth rate and suppressed the anchorage-independent growth of the SV40 cells. These results strongly support the view that DGAT participates in the regulation of membrane lipid synthesis and lipid signaling, thereby playing an important role in modulating cell growth properties. PMID- 14557276 TI - The SH3 domain-containing adaptor HIP-55 mediates c-Jun N-terminal kinase activation in T cell receptor signaling. AB - HIP-55 (hematopoietic progenitor kinase 1 (HPK1)-interacting protein of 55 kDa, also called SH3P7 and mAbp1) is a novel SH3 domain-containing protein. HIP-55 binds to actin filaments both in vitro and in vivo. HIP-55 activates HPK1 and c Jun N-terminal kinase (JNK), which are two important lymphocyte signaling molecules. Until now, the regulation and function of HIP-55 in T cell receptor (TCR) signaling were unknown. We found that HIP-55 was recruited to glycolipid enriched microdomains upon TCR stimulation, which indicates that HIP-55 is regulated by TCR signaling. HIP-55 interacted with ZAP-70, a critical protein tyrosine kinase in TCR signaling, and this interaction was induced by TCR signaling. ZAP-70 phosphorylated HIP-55 at Tyr-334 and Tyr-344 in vitro and in vivo, and the HIP-55 mutant (Y334F/Y344F) was not tyrosine-phosphorylated in stimulated T cells. To study its function in T cell activation, HIP-55-deficient Jurkat T cells were established using the RNA interference approach. In the HIP 55-deficient cells, TCR (but not UV)-stimulated JNK activation was decreased. Furthermore, the activation of HPK1, a known JNK upstream activator and HIP-55 interacting protein, was also decreased in the HIP-55-deficient cells. Our data reveal the regulation of HIP-55 during TCR signaling, and using a genetic approach, we demonstrate for the first time that HIP-55 plays a functional role in TCR signaling. PMID- 14557277 TI - The crystal structures of Klebsiella pneumoniae acetolactate synthase with enzyme bound cofactor and with an unusual intermediate. AB - Acetohydroxyacid synthase (AHAS) and acetolactate synthase (ALS) are thiamine diphosphate (ThDP)-dependent enzymes that catalyze the decarboxylation of pyruvate to give a cofactor-bound hydroxyethyl group, which is transferred to a second molecule of pyruvate to give 2-acetolactate. AHAS is found in plants, fungi, and bacteria, is involved in the biosynthesis of the branched-chain amino acids, and contains non-catalytic FAD. ALS is found only in some bacteria, is a catabolic enzyme required for the butanediol fermentation, and does not contain FAD. Here we report the 2.3-A crystal structure of Klebsiella pneumoniae ALS. The overall structure is similar to AHAS except for a groove that accommodates FAD in AHAS, which is filled with amino acid side chains in ALS. The ThDP cofactor has an unusual conformation that is unprecedented among the 26 known three dimensional structures of nine ThDP-dependent enzymes, including AHAS. This conformation suggests a novel mechanism for ALS. A second structure, at 2.0 A, is described in which the enzyme is trapped halfway through the catalytic cycle so that it contains the hydroxyethyl intermediate bound to ThDP. The cofactor has a tricyclic structure that has not been observed previously in any ThDP-dependent enzyme, although similar structures are well known for free thiamine. This structure is consistent with our proposed mechanism and probably results from an intramolecular proton transfer within a tricyclic carbanion that is the true reaction intermediate. Modeling of the second molecule of pyruvate into the active site of the enzyme with the bound intermediate is consistent with the stereochemistry and specificity of ALS. PMID- 14557278 TI - Effect of hypertension and hypertension-glucose intolerance on myocardial ischemic injury. AB - Systemic hypertension and type 2 diabetes mellitus are major risk factors for myocardial infarction. Yet, glucose intolerance, a prelude stage to type 2 diabetes, is associated with reduced infarct size. Since chronic hypertension adversely affects the myocardium, we tested the hypothesis that the coexistence of systemic hypertension and glucose intolerance reverses the cardioprotection associated with impaired glucose tolerance. Hearts from 9-month-old animals were subjected to a 40-minute occlusion of the left coronary artery followed by 2 hours of reperfusion. Before ischemia, similar values for the four experimental groups were observed for the coronary flow, heart rate, and maximum ventricular pressure. During the course of the ischemia-reperfusion insult, the two hypertensive groups displayed greater reductions in contractility than their normotensive counterparts. Infarct size was lower in the normotensive glucose intolerant rat than in the normotensive control rat. Surprisingly, the hypertrophied hearts of the hypertensive and hypertensive glucose-intolerant rats displayed reduced infarct size (P<0.05). However, raising the afterload pressure from 100 to 160 cm H2O increased infarct size in the two hypertensive groups. This narrowed the differential between the hypertensive glucose-intolerant (160 cm H2O) and the normotensive control (100 cm H2O) rats. Nonetheless, at the higher afterload pressure, infarct size was less in the hypertensive glucose intolerant rats than in their hypertensive counterparts. In conclusion, the impairment in contractile function despite the reduction in infarct size underscores the increased susceptibility of the hypertrophied, hypertensive heart to ischemic injury. Furthermore, exacerbation of cell death at elevated afterload pressure indicates the potential benefit of aggressive antihypertensive therapy. PMID- 14557280 TI - Role of endothelium-derived hyperpolarizing factor in human forearm circulation. AB - Endothelium-derived hyperpolarizing factor (EDHF) contributes to endothelium dependent relaxation of isolated arteries, but it is not known whether this also occurs in the case of humans in vivo. The present study examined the role of EDHF in human forearm circulation. Forearm blood flow (FBF) was measured by strain gauge plethysmography in 31 healthy, normal subjects (mean+/-SE age, 23+/-2 years; 24 men and 7 women). After oral administration of aspirin (486 mg), we infused NG-monomethyl-L-arginine (8 micromol/min for 5 minutes) into the brachial artery. We used tetraethylammonium chloride (TEA, 1 mg/min for 20 minutes), a KCa channel blocker, as an EDHF inhibitor, and nicorandil as a direct K+ channel opener. TEA significantly reduced FBF (P<0.05) but did not change systemic arterial blood pressure. Furthermore, TEA significantly inhibited the FBF increase in response to substance P (0.8, 1.6, 3.2, and 6.4 ng/min, n=8) and bradykinin (12.5, 25, 50, and 100 ng/min, n=8; both P<0.001), whereas it did not affect the FBF increase in response to acetylcholine (4, 8, 16, and 32 microg/min, n=8), sodium nitroprusside (0.4, 0.8, 1.6, and 3.2 microg/min, n=8), or nicorandil (0.128, 0.256, 0.512, and 1.024 mg/min, n=8). These results suggest that EDHF contributes substantially to basal forearm vascular resistance, as well as to forearm vasodilatation evoked by substance P and bradykinin in humans in vivo. PMID- 14557279 TI - Celiprolol activates eNOS through the PI3K-Akt pathway and inhibits VCAM-1 Via NF kappaB induced by oxidative stress. AB - Vascular cell adhesion molecule-1 (VCAM-1) and reactive oxygen species play critical roles in early atherogenesis, and nitric oxide (NO) is an important regulator of the cardiovascular system. Although celiprolol, a specific beta1 antagonist with weak beta2-agonistic action, stimulates endothelial nitric oxide synthase (eNOS) production, the mechanisms remain to be determined. Because it was recently reported that phosphatidylinositol 3-kinase (PI3K) and its downstream effector Akt are implicated in the activation of eNOS and that regulation of VCAM-1 expression is mediated via nuclear factor-kappaB (NF kappaB), we hypothesized that celiprolol activates phosphorylation of eNOS through the PI3K-Akt signaling pathway; that celiprolol modulates VCAM-1 expression, which is associated with inhibiting NF-kappaB phosphorylation; and that celiprolol suppresses NAD(P)H oxidase p22phox, p47phox, gp91phox, and nox1 expression in the left ventricle of deoxycorticosterone acetate (DOCA)-salt hypertensive rats. eNOS and Akt phosphorylation upregulated by celiprolol alone were suppressed by treatment with celiprolol plus wortmannin. Increased expression of VCAM-1, p22phox, p47phox, gp91phox, nox1, activated p65 NF-kappaB, c-Src, p44/p42 extracellular signal-regulated kinases, and their downstream effector p90 ribosomal S6 kinase phosphorylation in DOCA rats was inhibited by celiprolol. Celiprolol administration resulted in a significant improvement in cardiovascular remodeling and suppression of transforming growth factor-beta1 gene expression. In conclusion, celiprolol suppresses VCAM-1 expression because of inhibition of oxidative stress, NF-kappaB, and signal transduction, while increasing eNOS via stimulation of the PI3K-Akt signaling pathway and improving cardiovascular remodeling. PMID- 14557281 TI - Mechanisms mediating the vasoactive effects of the B1 receptors of bradykinin. AB - Bradykinin normally exerts its vasodilatory effect via the B2 receptor (B2R), but in this receptor's absence, the B1 receptor becomes expressed and activated. To explore the mechanism of B1R-mediated vasodilation, 8 groups of B2R gene-knockout mice received a 2-week infusion of a B1R antagonist (300 microg x kg(-1) x d(-1)) or vehicle (groups 1 and 2), B1R antagonist or vehicle plus NO inhibition with Nomega-nitro-L-arginine methyl ester (groups 3 and 4), B1R antagonist or vehicle plus cyclooxygenase inhibition with indomethacin (groups 5 and 6), or B1R antagonist or vehicle plus blockade of vasoconstricting prostaglandin (PG) H2 and thromboxane A2 (TxA2) with SQ29548 (groups 7 and 8). The B1R antagonist produced significant (P<0.05) blood pressure increases of 17.7+/-3.1 mm Hg in group 1 and 10.4+/-3 mm Hg in group 3, whereas their vehicle-treated respective control groups 2 and 4 had no significant blood pressure changes. Indomethacin abolished the capacity of the B1R antagonist to raise blood pressure, as did blockade of the receptors of PGH2 and TxA2. Injection with the B1R agonist produced a hypotensive response (12+/-1.3 mm Hg), which was further accentuated by TxA2 blockade (21.7+/-4.1 mm Hg). Analysis of B1R gene expression by reverse transcription-polymerase chain reaction (PCR) in cardiac and renal tissues revealed marked expression at baseline, with further upregulation by 1.5- to 2 fold after various manipulations. Expression of the TxA2 receptor gene in renal tissue by quantitative real-time PCR was significantly lower in mice treated with the B1R antagonist, consistent with increased levels of agonist for this receptor. The data confirm that the B1R becomes markedly expressed in the absence of B2R and suggest that it contributes to vasodilation by inhibiting a vasoconstricting product of the arachidonic acid cascade acting via the PGH2/TxA2 receptor. PMID- 14557282 TI - G-protein beta3-subunit gene 825T allele and hypertension: a longitudinal study in young grade I hypertensives. AB - The 825T allele of the GNB3 gene has been associated with essential hypertension and obesity in cross-sectional studies. We have therefore planned a longitudinal cohort study to assess whether the GNB3 825T allele is predictive of blood pressure increase in young subjects with grade I hypertension. We genotyped at the GNB3 825 locus 461 participants of the Hypertension and Ambulatory Recording Venetia Study (HARVEST) study (age, 18 to 45 years) at low cardiovascular risk, according to 1999 ISH/WHO criteria. The study end point was eligibility for antihypertensive medication, that is, progression to grade II hypertension during the first year of observation or office systolic blood pressure > or =150 mm Hg and/or office diastolic blood pressure > or =95 mm Hg in two later consecutive visits during follow-up. At baseline, there was no statistically significant difference among genotypes with respect to body mass index, blood pressure, and heart rate. During follow-up (mean, 4.7 years), 113 (51.1%) patients with CC genotype and 145 (60.4%) patients with TT/TC genotype reached the end point. According to survival analysis, the patients carrying the 825T allele had an increased risk of reaching the blood pressure end point (CI, 1.108 to 1.843; P=0.006). In young patients with grade I hypertension, the 825T allele is associated with increased risk of progression to more severe hypertension requiring antihypertensive therapy. The GNB3 825T allele may be considered a genetic marker of predisposition for hypertension. PMID- 14557283 TI - Blood pressure lowering and life expectancy based on a Markov model of cardiovascular events. AB - The life expectancy benefits of antihypertensive treatment, based on both systolic and diastolic blood pressure reduction, was estimated with a cardiovascular disease event Markov model with prospective data from 57 573 men and women. Seven patient states were defined, including (1) no cardiovascular disease, (2) stroke, (3) myocardial infarction, (4) revascularization, (5) history of cardiovascular disease, (6) noncardiovascular disease death, and (7) cardiovascular death. Risk functions were developed from gender-specific multivariate Cox proportional hazards models for primary events and age-, smoking , and diabetes-adjusted models for secondary events. At baseline we assumed (1) hypothetical pretreatment blood pressures of 160/95 or 150/90 mm Hg; (2) strategies A and B lower blood pressure by 20/13 and 13/8 mm Hg, respectively; and (3) baseline age of 35 years. For subjects initially at 160/95 mm Hg, those with antihypertensive treatment, antihypertensive treatment and diabetes, or antihypertensive treatment, diabetes, and currently smoking had corresponding gains in life expectancy of 2.43, 2.80, and 2.43 years for Strategy A. An initial blood pressure of 150/90 mm Hg resulted in similar gains. Compared with Strategy B, with blood pressure reductions of 13/8 mm Hg, Strategy A provided additional gains in life expectancy of 0.84, 0.99, and 0.87 years for those with antihypertensive treatment, antihypertensive treatment and diabetes, or antihypertensive treatment, diabetes, and currently smoking. The initial blood pressure level did not affect the magnitude of life expectancy gains for equivalent blood pressure reductions. Greater gains in life expectancy among hypertensive and diabetic women suggest that blood pressure lowering may yield greater benefits in selected subgroups. PMID- 14557284 TI - Short-term blood pressure, noradrenergic, and vascular effects of nocturnal home hemodialysis. AB - Long-term nocturnal hemodialysis, which uses longer and more frequent sessions than conventional hemodialysis, lowers clinic blood pressure and left ventricular mass. We tested the hypotheses that short-term nocturnal hemodialysis would (1) reduce ambulatory blood pressure; (2) cause peripheral vasodilation; (3) lower plasma norepinephrine concentration; and (4) improve the arterial response to reactive hyperemia (a marker of endothelium-dependent vasodilation). We studied 18 consecutive patients (age, 41+/-2; [mean+/-SEM]) before and 1 and 2 months after conversion from conventional (three 4-hour sessions per week) to nocturnal (six 8-hour sessions per week) hemodialysis. As the dialysis dose per session (Kt/V) increased from 1.24+/-0.06 to 2.04+/-0.08 after 2 months (P=0.02), symptomatic hypotension developed and most antihypertensive medications were withdrawn. Nocturnal hemodialysis nonetheless lowered 24-hour mean arterial pressure (from 102+/-3 to 90+/-2 mm Hg after 2 months; P=0.01), total peripheral resistance (from 1967+/-235 to 1499+/-191 dyne x s x cm(-5); P<0.01) and plasma norepinephrine (from 2.66+/-0.4 to 1.96+/-0.2 nmol; P=0.04). Endothelium dependent vasodilation could not be elicited during conventional hemodialysis ( 2.7+/-1.8%) but was restored (+8.0+/-1.0%; P=0.001) after 2 months of nocturnal hemodialysis. The brachial artery response to nitroglycerin also improved (from 6.9+/-2.8 to 15.7+/-1.6%; P<0.05). Nocturnal hemodialysis had no effect on weight or on stroke volume. Rapid reversal of these markers of adverse cardiovascular events with more intense hemodialysis may translate into improved outcome in this high-risk group of patients. PMID- 14557285 TI - Effect of shear stress on asymmetric dimethylarginine release from vascular endothelial cells. AB - We demonstrated recently that plasma concentrations of asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide (NO) synthase, are increased by high salt intake concomitantly with a decrease in plasma levels of NO in human hypertension. We investigated the effect of shear stress on ADMA release in 2 types of cells: transformed human umbilical vein endothelial cells (HUVECs; cell line ECV-304) and HUVECs. Exposure of ECV-304 cells and HUVECs to shear stress with the use of a cone-plate viscometer enhanced gene expression of protein arginine methyltransferase (PRMT-1), ADMA synthase. In HUVECs, the ratio of PRMT-1 to glyceraldehyde 3-phosphate dehydrogenase mRNA was increased by 2 fold by a shear stress of > or =15 dyne/cm2. A dominant-negative mutant of IkappaB kinase alpha and troglitazone at 8 micromol/L, an activator of peroxisome proliferator-activated receptor gamma, abolished the shear stress-induced increase in PRMT-1 gene expression in parallel with the blockade of nuclear factor (NF)-kappaB translocation into the nucleus. The activity of dimethylarginine dimethylaminohydrolase, the degradation enzyme of ADMA, was unchanged after shear stress < or =15 dyne/cm2 and was enhanced by 1.48+/-0.06 fold (P<0.05) by shear stress at 25 dyne/cm2. The release of ADMA was increased by 1.64+/-0.10-fold (P<0.05) by shear stress at 15 dyne/cm2 but was not affected by shear stress at 25 dyne/cm2. These results indicate that shear stress enhances gene expression of PRMT-1 and ADMA release via activation of the NF-kappaB pathway. Shear stress at higher magnitudes facilitates the degradation of ADMA, thus returning ADMA release levels to baseline. PMID- 14557286 TI - Cytoskeletal reorganization by mycophenolic acid alters mesangial cell migration and contractility. AB - Cytoskeleton alterations are a hallmark of mesangial cell activation during glomerulosclerosis. The aim of this study was to investigate whether mycophenolic acid (MPA) affects cytoskeletal organization and motility of human mesangial cells. Using the IP15 cell line, we found that treatment with 1 micromol/L MPA inhibited both receptor-dependent (angiotensin II) and receptor-independent (KCl) contractile responses, as well as serum-induced migration activity, suggesting alterations in the intracellular mechanisms that control mesangial cell motility. Immunofluorescence studies of MPA-treated cells provided evidence for decreased membrane disassembly/reassembly of alpha-smooth muscle actin and F-actin fibers, which was correlated with sustained quantitative and qualitative modifications of actin-associated proteins: calponin was overexpressed and became associated with actin fibers, whereas phosphorylation levels of cofilin and myosin light chain increased, suggesting both an activation of the mechanisms responsible for actin polymerization and an inhibition of actin-depolymerizing processes. These observations support a stabilizing effect of MPA on the mesangial actin cytoskeleton, which constitutes an additive action by which MPA, beyond its anti inflammatory, antiproliferative and antifibrotic properties, might protect against excessive mesangial activation in the context of various glomerulopathies and kidney transplantation. PMID- 14557287 TI - Clinical potential of the acyclic nucleoside phosphonates cidofovir, adefovir, and tenofovir in treatment of DNA virus and retrovirus infections. AB - The acyclic nucleoside phosphonates HPMPC (cidofovir), PMEA (adefovir), and PMPA (tenofovir) have proved to be effective in vitro (cell culture systems) and in vivo (animal models and clinical studies) against a wide variety of DNA virus and retrovirus infections: cidofovir against herpesvirus (herpes simplex virus types 1 and 2 varicella-zoster virus, cytomegalovirus [CMV], Epstein-Barr virus, and human herpesviruses 6, 7, and 8), polyomavirus, papillomavirus, adenovirus, and poxvirus (variola virus, cowpox virus, vaccinia virus, molluscum contagiosum virus, and orf virus) infections; adefovir against herpesvirus, hepadnavirus (human hepatitis B virus), and retrovirus (human immunodeficiency virus types 1 [HIV-1] and 2 [HIV-2], simian immunodeficiency virus, and feline immunodeficiency virus) infections; and tenofovir against both hepadnavirus and retrovirus infections. Cidofovir (Vistide) has been officially approved for the treatment of CMV retinitis in AIDS patients, tenofovir disoproxil fumarate (Viread) has been approved for the treatment of HIV infections (i.e., AIDS), and adefovir dipivoxil (Hepsera) has been approved for the treatment of chronic hepatitis B. Nephrotoxicity is the dose-limiting side effect for cidofovir (Vistide) when used intravenously (5 mg/kg); no toxic side effects have been described for adefovir dipivoxil and tenofovir disoproxil fumarate, at the approved doses (Hepsera at 10 mg orally daily and Viread at 300 mg orally daily). PMID- 14557289 TI - Babesia divergens, a bovine blood parasite of veterinary and zoonotic importance. AB - Babesia divergens is an intraerythrocytic protozoan parasite, transmitted by the tick Ixodes ricinus, and is the main agent of bovine babesiosis in Europe. It is not only a cause of significant loss to the cattle industry; it can also infect immunocompromised humans, causing medical emergencies characterized by rapid fulmination and parasitemias that may exceed 70%. The current emphasis in Europe on sustainable agriculture and extensification is likely to lead to an increase in vector tick populations with increased risk of infection. Despite the veterinary and zoonotic importance of this parasite, relatively little research has been carried out on B. divergens, and many questions regarding the parasite's epidemiology and the host's response remain unanswered. A better understanding of the species' biology and host-parasite interactions may lead to improved control mechanisms and new trends in vaccine and antibabesial drug development. This review provides the first comprehensive summary of B. divergens biology, including its morphology, life cycle, and host specificity, and the current state of knowledge of both human and bovine infections. PMID- 14557290 TI - Role of Toll-like receptors in pathogen recognition. AB - The innate immune system relies on a vast array of non-clonally expressed pattern recognition receptors for the detection of pathogens. Pattern recognition receptors bind conserved molecular structures shared by large groups of pathogens, termed pathogen-associated molecular patterns. The Toll-like receptors (TLRs) are a recently discovered family of pattern recognition receptors which show homology with the Drosophila Toll protein and the human interleukin-1 receptor family. Engagement of different TLRs can induce overlapping yet distinct patterns of gene expression that contribute to an inflammatory response. The TLR family is characterized by the presence of leucine-rich repeats and a Toll/interleukin-1 receptor-like domain, which mediate ligand binding and interaction with intracellular signaling proteins, respectively. Most TLR ligands identified so far are conserved microbial products which signal the presence of an infection, but evidence for some endogenous ligands that might signal other danger conditions has also been obtained. Molecular mechanisms for pathogen associated molecular pattern recognition still remain elusive but seem to be more complicated than initially anticipated. In most cases, direct binding of microbial ligands to TLRs still has to be demonstrated. Moreover, Drosophila TLRs bind endogenous ligands, generated through a proteolytic cascade in response to an infection. In the case of endotoxin, recognition involves a complex of TLR4 and a number of other proteins. Moreover, TLR heterodimerization further extends the spectrum of ligands and modulates the response towards specific ligands. The fact that TLR expression is regulated in both a cell type- and stimulus-dependent fashion further contributes to the complexity. PMID- 14557288 TI - Infections in patients with inherited defects in phagocytic function. AB - Patients with defects in phagocytic function are predisposed to intracellular microorganisms and typically have early dissemination of the infection. Recognition of the underlying disorder and aggressive antimicrobial therapy has been beneficial for the patients. Improved understanding of the pathophysiology has also affected patient management by allowing specific, targeted immunomodulatory intervention. The disorders described in this review are not common but have had a significant impact on our understanding of the role of phagocytic cells in host defense. Conversely, understanding the role of the neutrophil and macrophage in infection has benefited not just the patients described in this review but also other patients with similar disease processes. PMID- 14557291 TI - Prevention of cytomegalovirus disease in recipients of allogeneic stem cell transplants. AB - The main risk factors for cytomegalovirus (CMV) disease in recipients of allogeneic stem cell transplants (SCT) are recipient CMV seropositivity and acute graft-versus-host disease. Currently, two antiviral strategies, prophylactic or preemptive antiviral treatment, are used for prevention of CMV disease. Preemptive treatment is most favorable when short-term (14-day) treatment is applied. Several methods are available for monitoring of CMV reactivation. PCR based CMV DNA detection assays are the most sensitive methods; however, the clinical benefit of this high sensitivity is unclear. Even more, there is lack of clarity whether PCR tests can better be performed with plasma, whole blood, or peripheral blood leukocyte samples. Recovery of a CMV-specific CD8(+) cytotoxic-T lymphocyte (CTL) response is necessary for preventing CMV reactivation and disease. Reconstitution of absolute CMV-specific CTL counts to values above 10 x 10(6) to 20 x 10(6) CTLs/liter is associated with protection from CMV disease. In the near future, preemptive therapy might be withheld in patients with CMV reactivation who are shown to have adequate CMV-specific cytotoxic T-cell levels. Antiviral therapy with (val)acyclovir has been studied only as prophylactic treatment for prevention of CMV infection. High-dose oral valacyclovir is more effective than acyclovir when used in addition to preemptive treatment of CMV reactivation with ganciclovir or foscarnet. Three antiviral drugs have been tested for preemptive therapy of CMV reactivation and/or treatment of CMV disease. Although intravenous ganciclovir is considered the drug of choice, foscarnet has similar efficacy and less toxicity, especially hematologic toxicity. Cidofovir has not been tested extensively, but so far the results are disappointing. Oral valganciclovir for preemptive treatment of SCT recipients is currently being studied. In addition to antiviral therapy, adoptive immunotherapy with CMV-specific cytotoxic T cells as prophylactic or preemptive therapy is a very elegant strategy; however, generation of these cells is expensive and time consuming, and therefore the therapy is not available at every transplantation center. Magnetic selection of CMV-specific CD8(+) T cells from peripheral blood by using HLA class I-peptide tetramers may be very promising, making this strategy more accessible. PMID- 14557293 TI - Bacterial wall as target for attack: past, present, and future research. AB - When Bacteria, Archaea, and Eucarya separated from each other, a great deal of evolution had taken place. Only then did extensive diversity arise. The bacteria split off with the new property that they had a sacculus that protected them from their own turgor pressure. The saccular wall of murein (or peptidoglycan) was an effective solution to the osmotic pressure problem, but it then was a target for other life-forms, which created lysoymes and beta-lactams. The beta-lactams, with their four-member strained rings, are effective agents in nature and became the first antibiotic in human medicine. But that is by no means the end of the story. Over evolutionary time, bacteria challenged by beta-lactams evolved countermeasures such as beta-lactamases, and the producing organisms evolved variant beta-lactams. The biology of both classes became evident as the pharmaceutical industry isolated, modified, and produced new chemotherapeutic agents and as the properties of beta-lactams and beta-lactamases were examined by molecular techniques. This review attempts to fit the wall biology of current microbes and their clinical context into the way organisms developed on this planet as well as the changes arising since the work done by Fleming. It also outlines the scientific advances in our understanding of this broad area of biology. PMID- 14557294 TI - American canine hepatozoonosis. AB - American canine hepatozoonosis (ACH) is a tick-borne disease that is spreading in the southeastern and south-central United States. Characterized by marked leukocytosis and periosteal bone proliferation, ACH is very debilitating and often fatal. Dogs acquire infection by ingesting nymphal or adult Gulf Coast ticks (Amblyomma maculatum) that, in a previous life stage, ingested the parasite in a blood meal taken from some vertebrate intermediate host. ACH is caused by the apicomplexan Hepatozoon americanum and has been differentiated from Old World canine hepatozoonosis caused by H. canis. Unlike H. canis, which is transmitted by the ubiquitous brown dog tick (Rhipicephalus sanguineus), H. americanum is essentially an accidental parasite of dogs, for which Gulf Coast ticks are not favored hosts. The geographic portrait of the disease parallels the known distribution of the Gulf Coast tick, which has expanded in recent years. Thus, the endemic cycle of H. americanum involves A. maculatum as definitive host and some vertebrate intermediate host(s) yet to be identified. Although coyotes (Canis latrans) are known to be infected, it is not known how important this host is in maintaining the endemic cycle. This review covers the biology of the parasite and of the tick that transmits it and contrasts ACH with classical canine hepatozoonosis. Clinical aspects of the disease are discussed, including diagnosis and treatment, and puzzling epidemiologic issues are examined. Brief consideration is given to the potential for ACH to be used as a model for study of angiogenesis and of hypertrophic osteoarthropathy. PMID- 14557292 TI - Potential uses of probiotics in clinical practice. AB - Probiotics are defined as live microorganisms that, when administered in adequate amounts, confer a health benefit on the host. There is now mounting evidence that selected probiotic strains can provide health benefits to their human hosts. Numerous clinical trials show that certain strains can improve the outcome of intestinal infections by reducing the duration of diarrhea. Further investigations have shown benefits in reducing the recurrence of urogenital infections in women, while promising studies in cancer and allergies require research into the mechanisms of activity for particular strains and better designed trials. At present, only a small percentage of physicians either know of probiotics or understand their potential applicability to patient care. Thus, probiotics are not yet part of the clinical arsenal for prevention and treatment of disease or maintenance of health. The establishment of accepted standards and guidelines, proposed by the Food and Agriculture Organization of the United Nations and the World Health Organization, represents a key step in ensuring that reliable products with suitable, informative health claims become available. Based upon the evidence to date, future advances with single- and multiple-strain therapies are on the horizon for the management of a number of debilitating and even fatal conditions. PMID- 14557295 TI - Antigens and alternatives for control of Anaplasma marginale infection in cattle. AB - Anaplasmosis, a tick-borne cattle disease caused by the rickettsia Anaplasma marginale, is endemic in tropical and subtropical areas of the world. The disease causes considerable economic loss to both the dairy and beef industries worldwide. Analyses of 16S rRNA, groESL, and surface proteins have resulted in the recent reclassification of the order Rickettsiales. The genus Anaplasma, of which A. marginale is the type species, now also includes A. bovis, A. platys, and A. phagocytophilum, which were previously known as Ehrlichia bovis, E. platys, and the E. phagocytophila group (which causes human granulocytic ehrlichiosis), respectively. Live and killed vaccines have been used for control of anaplasmosis, and both types of vaccines have advantages and disadvantages. These vaccines have been effective in preventing clinical anaplasmosis in cattle but have not blocked A. marginale infection. Thus, persistently infected cattle serve as a reservoir of infective blood for both mechanical transmission and infection of ticks. Advances in biochemical, immunologic, and molecular technologies during the last decade have been applied to research of A. marginale and related organisms. The recent development of a cell culture system for A. marginale provides a potential source of antigen for the development of improved killed and live vaccines, and the availability of cell culture-derived antigen would eliminate the use of cattle in vaccine production. Increased knowledge of A. marginale antigen repertoires and an improved understanding of bovine cellular and humoral immune responses to A. marginale, combined with the new technologies, should contribute to the development of more effective vaccines for control and prevention of anaplasmosis. PMID- 14557298 TI - A Canadian agency for public health: if not now, when? PMID- 14557302 TI - Physician supply: future tense. PMID- 14557296 TI - Laboratory diagnosis of amebiasis. AB - The detection of Entamoeba histolytica, the causative agent of amebiasis, is an important goal of the clinical microbiology laboratory. To assess the scope of E. histolytica infection, it is necessary to utilize accurate diagnostic tools. As more is discovered about the molecular and cell biology of E. histolytica, there is great potential for further understanding the pathogenesis of amebiasis. Molecular biology-based diagnosis may become the technique of choice in the future because establishment of these protozoa in culture is still not a routine clinical laboratory process. In all cases, combination of serologic tests with detection of the parasite (by antigen detection or PCR) offers the best approach to diagnosis, while PCR techniques remain impractical in many developing country settings. The detection of amebic markers in serum in patients with amebic colitis and liver abscess appears promising but is still only a research tool. On the other hand, stool antigen detection tests offer a practical, sensitive, and specific way for the clinical laboratory to detect intestinal E. histolytica. All the current tests suffer from the fact that the antigens detected are denatured by fixation of the stool specimen, limiting testing to fresh or frozen samples. PMID- 14557303 TI - Word watch. PMID- 14557304 TI - Physician supply: future tense. PMID- 14557305 TI - Physician supply: future tense. PMID- 14557307 TI - Physician supply: future tense. PMID- 14557308 TI - The whiplash debate. PMID- 14557311 TI - Violence in Liberia. PMID- 14557312 TI - Acute hepatitis associated with levofloxacin in a patient with renal insufficiency. PMID- 14557297 TI - Current perspectives on ophthalmic mycoses. AB - Fungi may infect the cornea, orbit and other ocular structures. Species of Fusarium, Aspergillus, Candida, dematiaceous fungi, and Scedosporium predominate. Diagnosis is aided by recognition of typical clinical features and by direct microscopic detection of fungi in scrapes, biopsy specimens, and other samples. Culture confirms the diagnosis. Histopathological, immunohistochemical, or DNA based tests may also be needed. Pathogenesis involves agent (invasiveness, toxigenicity) and host factors. Specific antifungal therapy is instituted as soon as the diagnosis is made. Amphotericin B by various routes is the mainstay of treatment for life-threatening and severe ophthalmic mycoses. Topical natamycin is usually the first choice for filamentous fungal keratitis, and topical amphotericin B is the first choice for yeast keratitis. Increasingly, the triazoles itraconazole and fluconazole are being evaluated as therapeutic options in ophthalmic mycoses. Medical therapy alone does not usually suffice for invasive fungal orbital infections, scleritis, and keratitis due to Fusarium spp., Lasiodiplodia theobromae, and Pythium insidiosum. Surgical debridement is essential in orbital infections, while various surgical procedures may be required for other infections not responding to medical therapy. Corticosteroids are contraindicated in most ophthalmic mycoses; therefore, other methods are being sought to control inflammatory tissue damage. Fungal infections following ophthalmic surgical procedures, in patients with AIDS, and due to use of various ocular biomaterials are unique subsets of ophthalmic mycoses. Future research needs to focus on the development of rapid, species-specific diagnostic aids, broad-spectrum fungicidal compounds that are active by various routes, and therapeutic modalities which curtail the harmful effects of fungus- and host tissue-derived factors. PMID- 14557313 TI - Potential use of safer injecting facilities among injection drug users in Vancouver's Downtown Eastside. AB - BACKGROUND: The Vancouver Coastal Health Authority will initiate North America's first sanctioned safer injecting facility, as a pilot project, on Sept. 15, 2003. The analyses presented here were conducted to estimate the potential use of safer injecting facilities by local illicit injection drug users (IDUs) and to evaluate the potential impact of newly established Health Canada restrictions and current police activities on the use of the proposed facility. METHODS: During April and May 2003, we recruited active IDUs in Vancouver's Downtown Eastside to participate in a feasibility study. We used descriptive and univariate statistics to determine potential use of a safer injecting facility and to explore factors associated with willingness to use such a facility with and without federal restrictions and police presence. RESULTS: Overall, 458 street-recruited IDUs completed an interviewer-administered survey, of whom 422 (92%) reported a willingness to use a safer injecting facility. Those expressing willingness were more likely to inject in public (odds ratio [OR] 3.9, 95% confidence interval [CI] 1.9-8.0). When the restrictions in the Health Canada guidelines were considered, only 144 (31%) participants were willing to use a safer injecting facility. IDUs who inject alone were more likely (OR 1.8, 95% CI 1.0-3.1) and women were less likely (OR 0.6, 95% CI 0.4-0.9) to be willing to use a safer injecting facility operating under these restrictions. Only 103 (22%) of the participants said they would be willing to use a safer injecting facility if police were stationed near the entrance. INTERPRETATION: Most IDUs participating in this study expressed a willingness to use a safer injecting facility. However, willingness declined substantially when the IDUs were asked about using a facility operating under selected Health Canada restrictions and in the event that police were stationed near the entrance. PMID- 14557315 TI - Safer injecting facilities in Vancouver: considering issues beyond potential use. PMID- 14557314 TI - Incidence and estimated rates of residual risk for HIV, hepatitis C, hepatitis B and human T-cell lymphotropic viruses in blood donors in Canada, 1990-2000. AB - BACKGROUND: Since 1990, the Canadian Red Cross Society and Canadian Blood Services have been testing blood donors for hepatitis C virus (HCV) antibody and HCV nucleic acids and have supplemented HIV antibody testing with p24 antigen testing. We report trends in the incidence of blood-transmissible viral markers and estimates of the risk of undetected infection in donors over the last decade. METHODS: We extracted anonymous donor and blood-transmissible disease information from the Canadian Blood Services National Epidemiology Donor Database for 8.9 million donations from 2.1 million donors between June 1990 and December 2000. The risk of transfusion-transmitted infection (or "residual risk") refers to the chance that an infected donation escapes detection because of a laboratory test's window period (i.e., the time between infection and detection of the virus by that test). We determined the probability of residual contamination of a unit of blood after testing by using the incidence/window period model, which is based on the incidence of infection in repeat donors and the window period for each laboratory test. The viral markers evaluated in the study were HIV, HCV, hepatitis B virus (HBV) and human T-cell lymphotropic virus (HTLV). RESULTS: Except for HBV, the transmissible-disease rates of the other evaluated viruses decreased over the study period, with less of a decrease for HTLV. In 2000, the transmissible-disease-positive rate per 100 000 donations was 0.38 for HIV, 16.83 for HCV, 12.40 for HBV and 1.77 for HTLV. The residual risk of HIV, HCV and HTLV decreased over the study period; the residual risk of HBV fluctuated throughout the decade. The current residual risk per million donations is 0.10 for HIV, 0.35 for HCV, 13.88 for HBV and 0.95 for HTLV. INTERPRETATION: Except for HBV, the estimated risk of undetected infection (residual risk) has decreased over time. The rates of transmissible disease and the probability of undetected transmission of infection are at par with, if not lower than, those reported for other industrialized countries. PMID- 14557316 TI - Protection at the cutting edge: the case for central review of human gene transfer research. PMID- 14557317 TI - A practical approach to airway management in patients with SARS. PMID- 14557318 TI - Recommendations for screening and prevention of tuberculosis in patients with HIV and for screening for HIV in patients with tuberculosis and their contacts. PMID- 14557319 TI - Determining resuscitation preferences of elderly inpatients: a review of the literature. AB - Studies have shown that discussions with elderly hospital patients about cardiopulmonary resuscitation (CPR) preferences occur infrequently and have variable content. Our objective was to identify themes in the existing literature that could be used to increase the frequency and improve the quality of such discussions. We found that patients and families are familiar with the concept of CPR but have limited understanding of the procedure and overestimate its benefit. Most patients are interested in being involved in discussions about CPR and in sharing responsibility for decisions with physicians; however, older patients who participate in these discussions may have variable decision-making capacity. Physicians do not routinely discuss CPR with older patients, and patients do not initiate such discussions. When discussions do occur, the information provided to patients or families about resuscitation and its outcomes is not always consistent. Physicians should initiate CPR discussions, consider patients' levels of understanding and decision-making capacity, share responsibility for decisions where appropriate and involve the family where possible. Documentation of discussions and patient preferences may help to minimize misunderstandings and increase the stability of the decision during subsequent admissions to hospital. PMID- 14557320 TI - High-altitude decompression illness: case report and discussion. AB - Decompression illness (DCI) can occur in a variety of contexts, including scuba diving and flight in nonpressurized aircraft. It is characterized by joint pain, neurologic injury, and respiratory or constitutional symptoms. To prepare flight crews for accidental decompression events, the Canadian Armed Forces regularly conducts controlled and supervised depressurization exercises in specialized chambers. We present the cases of 3 Canadian Armed Forces personnel who successfully completed such decompression exercises but experienced DCI after they took a 3-hour commercial flight 6 hours after the completion of training. All 3 patients were treated in a hyperbaric oxygen chamber. The pathophysiology, diagnosis and management of DCI and the travel implications for military personnel who have undergone such training exercises are discussed. Although DCI is relatively uncommon, physicians may see it and should be aware of its presentation and treatment. PMID- 14557321 TI - Congenital abdominal wall defects. PMID- 14557322 TI - A 12-year-old girl with pharyngitis, meningitis and sinovenous thrombosis. PMID- 14557327 TI - Is it flu or SARS? MDs gear up for a difficult winter. PMID- 14557328 TI - New SARS recommendations to be released this month. PMID- 14557329 TI - Canadian media say DTC pharma ads the right Rx. PMID- 14557332 TI - Pharmacies, not MDs, should distribute medicinal marijuana: chair. PMID- 14557334 TI - Canada's approach to public health must be reinvented: SARS report. PMID- 14557336 TI - Will police presence deter use of drug injection site? PMID- 14557339 TI - All hospitals are not equal for treatment of patients with acute myocardial infarction. PMID- 14557341 TI - MADIT II (second Multicenter Automated Defibrillator Implantation Trial) debate: risk stratification, costs, and public policy. PMID- 14557340 TI - From vulnerable plaque to vulnerable patient: a call for new definitions and risk assessment strategies: Part II. AB - Atherosclerotic cardiovascular disease results in >19 million deaths annually, and coronary heart disease accounts for the majority of this toll. Despite major advances in treatment of coronary heart disease patients, a large number of victims of the disease who are apparently healthy die suddenly without prior symptoms. Available screening and diagnostic methods are insufficient to identify the victims before the event occurs. The recognition of the role of the vulnerable plaque has opened new avenues of opportunity in the field of cardiovascular medicine. This consensus document concludes the following. (1) Rupture-prone plaques are not the only vulnerable plaques. All types of atherosclerotic plaques with high likelihood of thrombotic complications and rapid progression should be considered as vulnerable plaques. We propose a classification for clinical as well as pathological evaluation of vulnerable plaques. (2) Vulnerable plaques are not the only culprit factors for the development of acute coronary syndromes, myocardial infarction, and sudden cardiac death. Vulnerable blood (prone to thrombosis) and vulnerable myocardium (prone to fatal arrhythmia) play an important role in the outcome. Therefore, the term "vulnerable patient" may be more appropriate and is proposed now for the identification of subjects with high likelihood of developing cardiac events in the near future. (3) A quantitative method for cumulative risk assessment of vulnerable patients needs to be developed that may include variables based on plaque, blood, and myocardial vulnerability. In Part I of this consensus document, we cover the new definition of vulnerable plaque and its relationship with vulnerable patients. Part II of this consensus document will focus on vulnerable blood and vulnerable myocardium and provide an outline of overall risk assessment of vulnerable patients. Parts I and II are meant to provide a general consensus and overviews the new field of vulnerable patient. Recently developed assays (eg, C-reactive protein), imaging techniques (eg, CT and MRI), noninvasive electrophysiological tests (for vulnerable myocardium), and emerging catheters (to localize and characterize vulnerable plaque) in combination with future genomic and proteomic techniques will guide us in the search for vulnerable patients. It will also lead to the development and deployment of new therapies and ultimately to reduce the incidence of acute coronary syndromes and sudden cardiac death. We encourage healthcare policy makers to promote translational research for screening and treatment of vulnerable patients. PMID- 14557342 TI - Cardiac memory and cortical memory: do learning patterns in neural networks impact on cardiac arrhythmias? AB - Memory is a property of diverse biological systems, including brain and heart. Studies in cortical neuronal networks have identified an increased sensitivity to infrequent (rare) stimulation patterns that can result in their achieving dominance over network firing. This adaptive behavior is applied to the heart in an attempt to explain the ability of pulmonary venous and other ectopic foci to achieve dominance over cardiac rhythm. Developmental changes in determinants of cardiac rhythm are explored as possible determinants of the range of rhythms expressed by the heart. By understanding the mechanisms for these behavior patterns, we may obtain new means for manipulating memory to return dysrhythmic hearts to normal sinus rhythm. PMID- 14557343 TI - Aldosterone blockade in patients with systolic left ventricular dysfunction. PMID- 14557344 TI - ACC/AHA/ESC guidelines for the management of patients with supraventricular arrhythmias--executive summary: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines and the European Society of Cardiology Committee for Practice Guidelines (Writing Committee to Develop Guidelines for the Management of Patients With Supraventricular Arrhythmias). PMID- 14557345 TI - Cardiology patient pages. Computed tomography scan and magnetic resonance imaging. PMID- 14557346 TI - Images in cardiovascular medicine. Assessment of cor triatriatum sinistrum by magnetic resonance imaging. PMID- 14557347 TI - Trends in stroke mortality. PMID- 14557348 TI - Arrhythmia monitoring through pacemakers. PMID- 14557349 TI - Morning blood pressure surge and the risk of stroke. PMID- 14557350 TI - Effect of the Asp298 variant of endothelial nitric oxide synthase on survival for patients with congestive heart failure. PMID- 14557351 TI - Angiotensin-converting enzyme inhibitor trandolapril does not affect C-reactive protein levels in myocardial infarction patients. PMID- 14557352 TI - Two better than one. PMID- 14557353 TI - Preserved alpha-adrenergic tone in the leg vascular bed of spinal cord-injured individuals. AB - BACKGROUND: Supraspinal sympathetic control of leg vascular tone is lost in spinal cord-injured individuals, but this does not result in a reduced leg vascular tone: Leg vascular resistance is even increased. The aim of this study was to assess the alpha-adrenergic contribution to the increased vascular tone in the lower extremity in patients without central sympathetic control of leg circulation. METHODS AND RESULTS: Upper-leg vascular resistance responses to local infusion of incremental doses of phentolamine (a competitive antagonist of the alpha-adrenoceptor) into the femoral artery were determined in 10 spinal cord injured individuals (SCI) and 8 healthy age-matched control subjects during local beta-adrenergic receptor blockade with propranolol. Basal leg vascular resistance was higher in SCI than in control subjects (41+/-6 arbitrary units [AU] versus 24+/-4 AU; P=0.034). The same accounts for minimal leg vascular resistance, assessed during reactive hyperemia, which was higher in SCI compared with control subjects (6.9+/-1.0 AU versus 2.5+/-0.2 AU; P<0.01). The maximal phentolamine induced reduction in leg vascular resistance normalized to each individual's minimal resistance did not differ between the groups (68+/-17% and 51+/-4% for SCI and control subjects, respectively; P>0.1). A decline in mean arterial pressure was observed in both groups with increasing dosage of phentolamine. In response, baroreceptor-mediated vasoconstriction was observed in the noninfused leg of the control subjects, whereas in SCI individuals this reaction was absent. CONCLUSIONS: These results indicate that the alpha-adrenoceptor-mediated vascular tone in the leg is preserved in spinal cord-injured individuals without sympathetic supraspinal control. PMID- 14557354 TI - Sirolimus-eluting stent implantation in ST-elevation acute myocardial infarction: a clinical and angiographic study. AB - BACKGROUND: Sirolimus-eluting stents (SES) have recently been proven to reduce restenosis and reintervention compared with bare stents. Safety and effectiveness of SES in acute myocardial infarction remain unknown. METHODS AND RESULTS: Since April 16, 2002, a policy of routine SES implantation has been instituted in our hospital, with no clinical or anatomic restrictions, as part of the RESEARCH (Rapamycin-Eluting Stent Evaluated At Rotterdam Cardiology Hospital) registry. During 6 months of enrollment, 96 patients with ST-elevation acute myocardial infarction underwent percutaneous recanalization and SES implantation; these patients comprise the study population. The incidence of major adverse cardiac events (death, nonfatal myocardial infarction, reintervention) was evaluated. Six month angiographic follow-up was scheduled per protocol. At baseline, diabetes mellitus was present in 12.5% and multivessel disease in 46.9%. Primary angioplasty was performed in 89 patients (92.7%). Infarct location was anterior in 41 (42.7%) of the cases, and 12 patients (12.5%) had cardiogenic shock. Postprocedural TIMI-3 flow was achieved in 93.3% of the cases. In-hospital mortality was 6.2%. One patient (1.1%) had reinfarction and target lesion reintervention the first day as a result of distal dissection and acute vessel occlusion. During follow-up (mean follow-up of 218+/-75 days), 1 patient died (1.1%), no patient had recurrent myocardial infarction, and there were no additional reinterventions. No early or late stent thromboses were documented. At angiographic follow-up (70%), late loss was -0.04+/-0.25, and no patient presented angiographic restenosis. CONCLUSIONS: In this study, sirolimus-eluting stent implantation for patients with ST-elevation acute myocardial infarction was safe without documented angiographic restenosis at 6 months. PMID- 14557355 TI - Catheter ablation for paroxysmal atrial fibrillation: segmental pulmonary vein ostial ablation versus left atrial ablation. AB - BACKGROUND: Segmental ostial catheter ablation (SOCA) to isolate the pulmonary veins (PVs) and left atrial catheter ablation (LACA) to encircle the PVs both may eliminate paroxysmal atrial fibrillation (PAF). The relative efficacy of these 2 techniques has not been directly compared. METHODS AND RESULTS: Of 80 consecutive patients with symptomatic PAF (age, 52+/-10 years), 40 patients underwent PV isolation by SOCA and 40 patients underwent LACA to encircle the PVs. During SOCA, ostial PV potentials recorded with a ring catheter were targeted. LACA was performed by encircling the left- and right-sided PVs 1 to 2 cm from the ostia and was guided by an electroanatomic mapping system; ablation lines also were created in the mitral isthmus and posterior left atrium. The mean procedure and fluoroscopy times were 156+/-45 and 50+/-17 minutes for SOCA and 149+/-33 and 39+/-12 minutes for LACA, respectively. At 6 months, 67% of patients who underwent SOCA and 88% of patients who underwent LACA were free of symptomatic PAF when not taking antiarrhythmic drug therapy (P=0.02). Among the variables of age, sex, duration and frequency of PAF, ejection fraction, left atrial size, structural heart disease, and the ablation technique, only an increased left atrial size and the SOCA technique were independent predictors of recurrent PAF. The only complication was left atrial flutter in a patient who underwent LACA. CONCLUSIONS: In patients undergoing catheter ablation for PAF, LACA to encircle the PVs is more effective than SOCA. PMID- 14557356 TI - Infarct remodeling after intracoronary progenitor cell treatment in patients with acute myocardial infarction (TOPCARE-AMI): mechanistic insights from serial contrast-enhanced magnetic resonance imaging. AB - BACKGROUND: Experimental and initial clinical studies suggest that transplantation of circulating blood- (CPC) or bone marrow-derived (BMC) progenitor cells may beneficially affect postinfarction remodeling processes after acute myocardial infarction (AMI). To relate functional characteristics of the infused cells to quantitative measures of outcome at 4-month follow-up, we performed serial contrast-enhanced MRI and assessed the migratory capacity of the transplanted progenitor cells immediately before intracoronary infusion. METHODS AND RESULTS: In 28 patients with reperfused AMI receiving either BMCs or CPCs into the infarct artery 4.7+/-1.7 days after AMI, serial contrast-enhanced MRI performed initially and after 4 months revealed a significant increase in global ejection fraction (from 44+/-10% to 49+/-10%; P=0.003), a decrease in end systolic volume (from 69+/-26 to 60+/-28 mL; P=0.003), and unchanged end diastolic volumes (122+/-34 versus 117+/-37 mL; P=NS). Infarct size, measured as late enhancement (LE) volume, decreased significantly, from 46+/-32 to 37+/-28 mL (P<0.05). There was a significant correlation between the reduction in LE volume and global ejection fraction improvement. The migratory capacity of transplanted cells as assessed ex vivo toward a gradient of vascular endothelial growth factor for CPCs and stromal cell derived factor-1 for BMCs was closely correlated with the reduction of LE volume. By multivariate analysis, migratory capacity remained the most important independent predictor of infarct remodeling. CONCLUSIONS: Analysis of serial contrast-enhanced MRI suggests that intracoronary infusion of adult progenitor cells in patients with AMI beneficially affects postinfarction remodeling processes. The migratory capacity of the infused cells is a major determinant of infarct remodeling, disclosing a causal effect of progenitor cell therapy on regeneration enhancement. PMID- 14557357 TI - Predictors of improved renal function after percutaneous stent-supported angioplasty of severe atherosclerotic ostial renal artery stenosis. AB - BACKGROUND: Percutaneous stent-supported angioplasty is a treatment option for atherosclerotic ostial renal artery stenosis. Improvement of renal function by such intervention, however, is controversial and thought to be limited to specific subsets, such as nondiabetic patients and bilateral stenoses. In this prospective study, we investigated predictors for improvement of renal function and blood pressure after renal artery stent placement. METHODS AND RESULTS: The study included 215 consecutive patients with ostial renal artery stenosis of > or =70% diameter stenosis undergoing stent-supported angioplasty. The primary end point was decrease in serum creatinine concentration at 1 year; the secondary end point, decrease in average mean arterial blood pressure assessed by 24-hour monitoring. One-year follow-up was complete in 191 surviving patients. In 52% (99/191) of the patients, serum creatinine concentration decreased during 1-year follow-up. Median serum creatinine concentration dropped significantly from 1.21 mg/dL (quartiles: 0.92, 1.60 mg/dL) at baseline to 1.10 mg/dL (quartiles: 0.88, 1.50 mg/dL) at 1 year (P=0.047). On average, mean arterial blood pressure decreased significantly, from 102+/-12 mm Hg (mean+/-SD) at baseline to 92+/-10 mm Hg at 1 year (P<0.001). Significant independent predictors of improved renal function were baseline serum creatinine (odds ratio [95% CI], 2.58 [1.35 to 4.94], P=0.004) and left ventricular function (OR 1.51 [1.04 to 2.21], P=0.032). Female sex, high baseline mean blood pressure, and normal renal parenchymal thickness were independent predictors for decreased mean blood pressure. CONCLUSIONS: Stent-supported angioplasty for severe ostial renal artery stenosis improves renal function and blood pressure in a broader spectrum of patients than previously thought. PMID- 14557358 TI - Dynamic regulation of sodium/calcium exchange function in human heart failure. AB - BACKGROUND: Sarcolemmal Na/Ca exchange (NCX) regulates cardiac Ca and contractility. NCX function during the cardiac cycle is determined by intracellular [Ca] and [Na] ([Ca]i, and [Na]i) and membrane potential (Em), which all change in human heart failure (HF). Therefore, changes in NCX function may contribute to abnormal Ca regulation in human HF. METHODS AND RESULTS: We assessed the cellular bases of differences in NCX function in ventricular myocytes from failing (F) and nonfailing (NF) human hearts. Allosteric activation of NCX by [Ca]i was comparable in F and NF myocytes (K1/2=150+/-31 nmol/L, n=7). The steady-state relation between [Ca]i and NCX current (INCX) was used to infer the local submembrane [Ca]i ([Ca]sm) that is sensed by NCX dynamically during the action potential (AP) and Ca transient (37 degrees C). This involved "tail" INCX measurement during abrupt repolarization of APs and Ca transients, where peak inward INCX indicates [Ca]sm. This allows inference of the direction of Ca transport by the NCX during the AP. In NF myocytes, NCX extrudes Ca for most of the AP. Three factors shift the direction of NCX-mediated Ca transport (to favor more Ca influx) in F versus NF myocytes, as follows: (1) reduced [Ca]sm, (2) prolonged AP duration, and (3) elevated [Na]i. CONCLUSIONS: These results show that Ca entry through NCX may limit systolic dysfunction due to reduced sarcoplasmic reticulum Ca stores in HF but could contribute to slow decay of the [Ca]i transient and to diastolic dysfunction. PMID- 14557359 TI - Blood oxygen level-dependent magnetic resonance imaging in patients with stress induced angina. AB - BACKGROUND: Blood oxygen level-dependent (BOLD) MRI reflects tissue oxygenation and may be useful for the detection of myocardial ischemia in patients with suspected coronary artery disease. METHODS AND RESULTS: We studied 25 patients with stress-induced angina using a T2*-sensitive echo planar imaging sequence before and during adenosine in a single-slice approach. BOLD-MRI results were compared with quantitative angiography and adenosine thallium single-photon emission computed tomography (SPECT). Although image quality was variable because of artifacts, no data were excluded from the analysis. During adenosine, a mean signal intensity decrease was observed for myocardial segments related to coronary stenoses >75%. On average, a nonsignificant increase was observed in the other segments. The angiographically determined stenosis was correlated with BOLD MRI results. Including all segments and using BOLD-MRI signal intensity increase cutoff value of 1.2%, BOLD-MRI had a sensitivity of 88% and a specificity of 47% to correctly classify severe stenoses. Adenosine thallium SPECT data from distal segments of the same coronary territory were also correlated with BOLD-MRI. However, variability was substantial. CONCLUSIONS: In patients with stress induced angina, adenosine BOLD-MRI detects myocardial ischemia in myocardial segments related to severe coronary stenoses. Its potential will increase with additional improvement of spatial coverage and image quality. PMID- 14557361 TI - Electrophysiological and electrocardiographic characteristics of focal atrial tachycardia originating from the pulmonary veins: acute and long-term outcomes of radiofrequency ablation. AB - BACKGROUND: The objective of this study was to describe the electrophysiological characteristics, anatomic distribution, and long-term outcome after focal ablation (RFA) of pulmonary vein (PV) atrial tachycardia (AT). Both atrial fibrillation (AF) and AT may be due to a rapidly firing focus in the PVs. Whether these represent two aspects of the same process is unknown. METHODS AND RESULTS: Twenty-seven patients with 28 PV(16%) ATs of a consecutive series of 172 undergoing RFA for focal AT are reported. The mean age was 39+/-16 years, with symptoms for 9+/-14 years resistant to 1.7+/-0.8 medications. AT occurred spontaneously or with isoproterenol in all patients and was not inducible with PES in any. The distribution of PV ATs was right superior PV, 11; left superior PV, 11; left inferior PV, 5; and right inferior PV, 1; 26of 28 foci (93%) were ostial. RFA was successful in 28 of 28 PV ATs acutely. RFA was focal in 25 of 28, with PV isolation of a single target vein in 3. There were 4 recurrences at a mean of 3.3 months. Repeat RFA was performed in all 4 and successful in 3 of 4. All but one recurrence occurred from the same site. Long-term success was achieved in 26 of 27 (96%) patients at mean follow-up of 25+/-22 months. No patients have had subsequent development of AF or AT from a different site. CONCLUSIONS: PV AT has a distribution similar to PV AF, with a propensity to upper veins. However, the majority of foci are ostial, and only a small percentage occur from deep in the PV. Focal RFA is associated with high long-term success, with freedom from both AT from other sites and from AF. PV AT is a localized process and therefore may be different from PV AF. PMID- 14557360 TI - Body mass index: a risk factor for unstable angina and myocardial infarction in patients with angiographically confirmed coronary artery disease. AB - BACKGROUND: In patients with coronary artery disease (CAD), acute thrombosis frequently occurs in coronary arteries with only mild or moderate stenoses. Obesity increases the risk of atherosclerosis, but it is not known whether it also increases the risk of coronary thrombosis. We hypothesized that body mass index (BMI) might be an independent predictor of an acute coronary syndrome in patients with established coronary atherosclerosis. METHODS AND RESULTS: Of 504 patients undergoing coronary angiography, those with evidence of >10% coronary artery stenoses were divided into 2 groups, with either stable (n=226) or unstable CAD (unstable angina or myocardial infarction; n=156). After adjusting for other risk factors (age, gender, blood pressure, lipid levels, insulin resistance, leptin, fibrinogen, C-reactive protein (CRP), CAD severity on angiography, smoking status, and a history of myocardial infarction or hypertension), BMI had a significant independent association with an acute coronary syndrome, with an odds ratio of 1.49 (P=0.014). This positive relation between BMI and the risk of acute coronary events was evident for even mildly elevated BMI values. Multivariate analysis also showed that CRP and the number of coronary lesions were independent predictors of risk of an acute coronary event. CONCLUSIONS: In patients with established coronary atherosclerosis, BMI, as well as CRP and number of coronary lesions, are independently associated with acute coronary syndromes. There is evidence of increased risk even at mildly elevated BMI levels. PMID- 14557362 TI - Joint effects of C-reactive protein and glycated hemoglobin in predicting future cardiovascular events of patients with advanced atherosclerosis. AB - BACKGROUND: C-reactive protein (CRP) and glycohemoglobin (HbA1c) are established risk factors for the development of cardiovascular disease. We investigated the joint effects of these parameters on cardiovascular outcome of patients with advanced atherosclerosis. METHODS AND RESULTS: We studied 454 patients with advanced atherosclerosis (median age, 69 years; 264 male). Cardiovascular risk profile, high-sensitivity CRP (hs-CRP), and HbA1c were obtained at baseline, and patients were followed for a median of 21 months (interquartile range, 13 to 26) for the occurrence of major adverse cardiovascular events (MACE) (myocardial infarction, percutaneous coronary interventions, coronary artery bypass graft, carotid revascularization, stroke, and death). We observed 166 MACE in 128 patients (28%). Cumulative event-free survival rates at 6, 12, and 24 months were 91%, 85%, and 73%, respectively. Adjusted hazard ratios for the occurrence of MACE according to increasing quartiles of hs-CRP and HbA1c were 1.35 (P=0.31), 1.90 (P=0.026) and 2.13 (P=0.007), and 1.40 (P=0.26), 1.81 (P=0.059), and 2.36 (P=0.023), respectively, compared with the lowest quartiles. Considering both parameters jointly, we found that patients with hs-CRP >0.44 mg/dL and HbA1c >6.2% (upper quartiles) were at highest risk for MACE, with each parameter adding to the prognostic information of the other. CONCLUSIONS: Inflammation, indicated by hs-CRP, and hyperglycemia, indicated by HbA1c, jointly contribute to the cardiovascular risk of patients with advanced atherosclerosis. Patients with both hs-CRP and HbA1c in the upper quartiles (>0.44 mg/dL and >6.2%, respectively) are at particularly high risk for poor cardiovascular outcome. PMID- 14557363 TI - Prognostic significance of small-artery structure in hypertension. AB - BACKGROUND: The presence of structural alterations in the microcirculation may be considered an important mechanism of organ damage; however, it is not currently known whether structural alterations of small arteries may predict fatal and nonfatal cardiovascular events. METHODS AND RESULTS: One hundred twenty-eight patients were included in the present study. There were 59 patients with essential hypertension, 17 with pheochromocytoma, 20 with primary aldosteronism, 12 with renovascular hypertension, and 20 normotensive patients with non-insulin dependent diabetes mellitus. All subjects were submitted to a biopsy of subcutaneous fat. Small resistance arteries were dissected and mounted on an isometric myograph, and the tunica media-to-internal lumen ratio (M/L) was measured. The subjects were reevaluated after an average follow-up time of 5.4 years. Thirty-seven subjects had a documented fatal or nonfatal cardiovascular event (5.32 events/100 patients per year). In the subcutaneous small arteries of subjects with cardiovascular events, a smaller internal diameter and a clearly greater M/L was observed. Our subjects were subdivided according to the presence of an M/L greater or smaller than the mean and median values observed in the whole population (0.098) or mean value +2 SD of our normal subjects (0.11). Life table analyses showed a significant difference in event-free survival between the subgroups. Cox's proportional hazard model, considering all known cardiovascular risk factors, indicated that only pulse pressure (P=0.009) and M/L (P<0.0001) were significantly associated with the occurrence of cardiovascular events. CONCLUSIONS: Our results strongly indicate a relevant prognostic role of structural alterations in small resistance arteries of a high-risk population. PMID- 14557364 TI - Gadolinium cardiovascular magnetic resonance predicts reversible myocardial dysfunction and remodeling in patients with heart failure undergoing beta-blocker therapy. AB - BACKGROUND: In some patients with heart failure, beta-blockers can improve left ventricular (LV) function and reduce morbidity and mortality. We hypothesized that gadolinium-enhanced cardiovascular magnetic resonance imaging (CMR) can predict reversible myocardial dysfunction and remodeling in heart failure patients treated with beta-blockers. METHODS AND RESULTS: Forty-five patients with chronic heart failure underwent CMR. Contrast imaging using gadolinium was performed to obtain high-resolution spatial maps of myocardial scarring and viability. Cine imaging was performed to assess LV function and morphology and was repeated in 35 patients after 6 months of beta-blockade. Gadolinium CMR demonstrated scarring in 30 of 45 patients (67%). Scarring was found in 100% of patients with ischemic cardiomyopathy (28 of 28) but in only 12% with nonischemic cardiomyopathy (2 of 17). In the 35 patients who were maintained on beta-blockers and had a second study, there was an inverse relation between the extent of scarring at baseline and the likelihood of contractile improvement 6 months later (P<0.001). For instance, contractility improved in 56% (674 of 1207) of regions with no scarring but in only 3% with >75% scarring (8 of 232). Multivariate analysis showed that the amount of dysfunctional but viable myocardium by CMR was an independent predictor of the change in ejection fraction (P=0.01), mean wall motion score (P=0.0007), LV end-diastolic volume index (P=0.007), and LV end systolic volume index (P< or =0.0001). CONCLUSIONS: For heart failure patients treated with beta-blockers, gadolinium-enhanced CMR predicts the response in LV function and remodeling. PMID- 14557365 TI - Ras induces vascular smooth muscle cell senescence and inflammation in human atherosclerosis. AB - BACKGROUND: Vascular cells have a finite cell lifespan and eventually enter an irreversible growth arrest, cellular senescence. The functional changes associated with cellular senescence are thought to contribute to human aging and age-related vascular disorders. Ras, an important signaling molecule involved in atherogenic stimuli, is known to promote aging in yeast and cellular senescence in primary human fibroblasts. The aim of this study was to investigate the role of Ras-induced vascular smooth muscle cell (VSMC) senescence in atherogenesis. METHODS AND RESULTS: We introduced an activated ras allele (H-rasV12) into human VSMCs using retroviral infection. Introduction of H-rasV12 induced a growth arrest with phenotypic characteristics of cellular senescence, such as enlarged cell shapes and increases in expression of cyclin-dependent kinase inhibitors and senescence-associated beta-galactosidase (SA-beta-gal) activity. Activation of Ras drastically increased expression of proinflammatory cytokines, in part through extracellular signal-regulated kinase activation. To determine whether Ras activation induces cellular senescence in vivo, we transduced the adenoviral vector encoding H-rasV12 into rat carotid arteries injured by a balloon catheter. Introduction of Ras into the arteries enhanced vascular inflammation and senescence compared with mock-infected injured arteries. Moreover, SA-beta-gal positive VSMCs were detected in the intima of advanced human atherosclerotic lesions and exhibited increased levels of extracellular signal-regulated kinase activity and proinflammatory cytokine expression. CONCLUSIONS: Our results suggest that atherogenic stimuli mediated by Ras induce VSMC senescence and vascular inflammation, thereby contributing to atherogenesis. This novel mechanism of atherogenesis may provide insights into a new antisenescence treatment for atherosclerosis. PMID- 14557366 TI - Dynamics of the pulmonary venous flow in the fetus and its association with vascular diameter. AB - BACKGROUND: The usual positioning of the Doppler sample volume to assess fetal pulmonary vein flow is in the distal portion of the vein, where the vessel diameter is maximal. This study was performed to test the association of the pulmonary vein pulsatility index (PVPI) with the vessel diameter. METHODS AND RESULTS: Twenty-three normal fetuses (mean gestational age, 28.6+/-5.3 weeks) were studied by Doppler echocardiography. Pulmonary right upper vein flow was assessed adjacent to the venoatrial junction ("distal" position) and in the middle of the vein ("proximal" position). The vessel diameter was measured by 2D echocardiography with power Doppler, and the PVPI was obtained by the ratio (maximal velocity [systolic or diastolic peak]-minimal velocity [presystolic peak])/mean velocity. The statistical analysis used t test and exponential correlation studies. Mean distal diameter was 0.33+/-0.10 cm (0.11 to 0.57 cm), and mean proximal diameter was 0.16+/-0.08 cm (0.11 to 0.25 cm) (P<0.0001). Mean distal PVPI was 0.84+/-0.21 (0.59 to 1.38), and mean proximal PVPI was 2.09+/ 0.59 (1.23 to 3.11) (P<0.0001). Exponential inverse correlation between pulmonary vein diameter and pulsatility index was highly significant (P<0.0001), with a determination coefficient of 0.439. CONCLUSIONS: In the normal fetus, the pulmonary venous flow pulsatility decreases from the lung to the heart, and this parameter is inversely correlated to the diameter of the pulmonary vein, which increases from its proximal to its distal portion. This study emphasizes the importance of the correct positioning of the Doppler sample volume, adjacent to the venoatrial junction, to assess pulmonary venous flow dynamics. PMID- 14557367 TI - Thromboxane A2 regulates vascular tone via its inhibitory effect on the expression of inducible nitric oxide synthase. AB - BACKGROUND: Circulatory failure in sepsis arises from vascular hyporesponsiveness, in which nitric oxide (NO) derived from inducible NO synthase (iNOS) plays a major role. Details of the cross talk between thromboxane (TX) A2 and the iNOS-NO system, however, remain unknown. We intended to clarify the role of TXA2, via the cross talk, in vascular hyporesponsiveness. METHODS AND RESULTS: We examined cytokine-induced iNOS expression and NO production in cultured vascular smooth muscle cells (VSMCs) and cytokine-induced hyporesponsiveness of the aorta from mice lacking the TXA2 receptor (TP-/- mice). The cytokine-induced iNOS expression and NO production observed in wild-type VSMCs were significantly augmented in TP-/- VSMCs, indicating an inhibitory effect of endogenous TXA2 on iNOS expression. Furthermore, in indomethacin-treated wild-type VSMCs, U-46619, a TP agonist, inhibited cytokine-induced iNOS expression and NO production in a concentration-dependent manner, effects absent from TP-/- VSMCs. In an ex vivo system, the cytokine-induced hyporesponsiveness of aortas to phenylephrine was significantly augmented in TP-/- aorta but was almost completely canceled by aminoguanidine, an iNOS inhibitor. Accordingly, cytokine-induced NO production was significantly higher in TP-/- aorta than in wild-type aorta. Moreover, U 46619 significantly suppressed lipopolysaccharide-induced NO production in vivo only in wild-type mice. CONCLUSIONS: These results suggest that TXA2 has a protective role against the development of vascular hyporesponsiveness via its inhibitory action on the iNOS-NO system under pathological conditions such as sepsis. PMID- 14557368 TI - Aldosterone regulates vascular reactivity: short-term effects mediated by phosphatidylinositol 3-kinase-dependent nitric oxide synthase activation. AB - BACKGROUND: There is increasing evidence for rapid nongenomic effects of aldosterone. Therefore, we studied the immediate effects of aldosterone on vascular reactivity in rat aortic ring segments and on endothelial and vascular smooth muscle cellular responses. METHODS AND RESULTS: In endothelium-intact ring segments, aldosterone attenuated phenylephrine-mediated constriction (maximal reduction, 25+/-4% below control phenylephrine-mediated constriction). In contrast, in endothelium-denuded vessels, aldosterone mediated a monophasic dose dependent enhancement of vasoconstrictor response. In endothelial cells, aldosterone caused a phosphatidylinositol 3-kinase (PI3K)-dependent increase in nitric oxide synthase activity as well as PI3K-dependent activation of extracellular signal-regulated kinase 1/2 and p70 S6 kinase. CONCLUSIONS: Overall, these data support a novel effect of aldosterone on vascular endothelial and smooth muscle cell function. These rapid effects of aldosterone might be important in both the short- and long-term regulation of peripheral vascular resistance. Furthermore, in the setting of endothelial dysfunction, alterations in aldosterone's short-term vascular responses might contribute to its pathophysiological effects in cardiovascular disease. PMID- 14557369 TI - Activation of peroxisome proliferator-activated receptor-alpha protects the heart from ischemia/reperfusion injury. AB - BACKGROUND: Peroxisome proliferator-activated receptor-alpha (PPAR-alpha) is expressed in the heart and regulates genes involved in myocardial fatty acid oxidation (FAO). The role of PPAR-alpha in acute ischemia/reperfusion myocardial injury remains unclear. METHODS AND RESULTS: The coronary arteries of male mice were ligated for 30 minutes. After reperfusion for 24 hours, ischemic and infarct sizes were determined. A highly selective and potent PPAR-alpha agonist, GW7647, was administered by mouth for 2 days, and the third dose was given 1 hour before ischemia. GW7647 at 1 and 3 mg x kg(-1) x d(-1) reduced infarct size by 28% and 35%, respectively (P<0.01), and myocardial contractile dysfunction was also improved. Cardioprotection by GW7647 was completely abolished in PPAR-alpha-null mice. Ischemia/reperfusion downregulated mRNA expression of cardiac PPAR-alpha and FAO enzyme genes, decreased myocardial FAO enzyme activity and in vivo cardiac fat oxidation, and increased serum levels of free fatty acids. All of these changes were reversed by GW7647. Moreover, GW7647 attenuated ischemia/reperfusion-induced release of multiple proinflammatory cytokines and inhibited neutrophil accumulation and myocardial expression of matrix metalloproteinases-9 and -2. Furthermore, GW7647 inhibited nuclear factor-kappaB activation in the heart, accompanied by enhanced levels of inhibitor-kappaBalpha. CONCLUSIONS: Activation of PPAR-alpha protected the heart from reperfusion injury. This cardioprotection might be mediated through metabolic and antiinflammatory mechanisms. This novel effect of the PPAR-alpha agonist could provide an added benefit to patients treated with PPAR-alpha activators for dyslipidemia. PMID- 14557371 TI - Effects of pulmonary insufficiency on biventricular function in the developing heart of growing swine. AB - BACKGROUND: This study was conducted to determine the effects of chronic pulmonary insufficiency (PI) on right (RV) and left (LV) ventricular function in young growing swine. METHODS AND RESULTS: Six PI and 5 control animals were studied. PI was induced by transcatheter placement of stents across the pulmonary valve. Indices of systolic function (ejection fraction, cardiac output, and cardiac functional reserve), diastolic function (compliance), and myocardial contractility (the slope of the relationship of end-systolic pressure versus end systolic volume [Emax] and the slope of the dP/dtmax-end-diastolic volume relationship [MdP/dt]) were assessed within 2 days of intervention and 3 months later. MRI was used to quantify PI and ventricular volumes. Conductance catheter techniques were used to obtain indices of contractility and diastolic compliance from pressure-volume relations at rest and under dobutamine infusion. In the PI group, pulmonary regurgitant fraction was 49.2+/-5.9% at 3-month follow-up. RV cardiac functional reserve was limited, diastolic function was preserved, and myocardial contractility was altered (Emax=2.6+/-0.3 mm Hg/mL for the PI group versus 3.5+/-0.4 mm Hg/mL for control; P<0.01). LV cardiac functional reserve was limited, ventricular compliance decreased, and myocardial contractility was preserved. CONCLUSIONS: In the young developing heart, chronic PI alters biventricular systolic function, RV myocardial contractility, and LV diastolic performance. PMID- 14557370 TI - Molecular imaging of angiogenesis in early-stage atherosclerosis with alpha(v)beta3-integrin-targeted nanoparticles. AB - BACKGROUND: Angiogenesis is a critical feature of plaque development in atherosclerosis and might play a key role in both the initiation and later rupture of plaques that lead to myocardial infarction and stroke. The precursory molecular or cellular events that initiate plaque growth and that ultimately contribute to plaque instability, however, cannot be detected directly with any current diagnostic modality. METHODS AND RESULTS: Atherosclerosis was induced in New Zealand White rabbits fed 1% cholesterol for approximately 80 days. alpha(v)beta3-Integrin-targeted, paramagnetic nanoparticles were injected intravenously and provided specific detection of the neovasculature within 2 hours by routine magnetic resonance imaging (MRI) at a clinically relevant field strength (1.5 T). Increased angiogenesis was detected as a 47+/-5% enhancement in MRI signal averaged throughout the abdominal aortic wall among rabbits that received alpha(v)beta3-targeted, paramagnetic nanoparticles. Pretreatment of atherosclerotic rabbits with alpha(v)beta3-targeted, nonparamagnetic nanoparticles competitively blocked specific contrast enhancement of the alpha(v)beta3-targeted paramagnetic agent. MRI revealed a pattern of increased alpha(v)beta3-integrin distribution within the atherosclerotic wall that was spatially heterogeneous along both transverse and longitudinal planes of the abdominal aorta. Histology and immunohistochemistry confirmed marked proliferation of angiogenic vessels within the aortic adventitia, coincident with prominent, neointimal proliferation among cholesterol-fed, atherosclerotic rabbits in comparison with sparse incidence of neovasculature in the control animals. CONCLUSIONS: This molecular imaging approach might provide a method for defining the burden and evolution of atherosclerosis in susceptible individuals as well as responsiveness of individual patients to antiatherosclerotic therapies. PMID- 14557372 TI - Reversal of atrial mechanical dysfunction after cardioversion of atrial fibrillation: implications for the mechanisms of tachycardia-mediated atrial cardiomyopathy. AB - BACKGROUND: Atrial mechanical "stunning" develops after cardioversion of atrial fibrillation (AF) and is implicated in the genesis of thromboembolic complications. However, the mechanisms responsible for this phenomenon are poorly understood. Whether atrial mechanical dysfunction caused by AF can be reversed by pacing at increased rates or by pharmacological agents is unknown. METHODS AND RESULTS: Twenty-six patients with AF undergoing cardioversion were dichotomized prospectively on the basis of the duration of arrhythmia as short-duration (1 to 6 months) or long-duration (> or =3 years) AF. Left atrial appendage emptying velocities (LAAEVs) and spontaneous echocardiographic contrast (LASEC) were assessed by transesophageal echocardiography during AF, after reversion to sinus rhythm, during atrial pacing at cycle lengths of 750 to 250 ms, after a postpacing pause, and with isoproterenol. In patients with short-duration AF, LAAEV decreased (42.0+/-2.7 to 18.5+/-2.0 cm/s; P<0.0001) and LASEC increased (0.9+/-0.3 to 2.2+/-0.3; P<0.01) with termination of AF; pacing increased LAAEV (48.3+/-4.1 cm/s; P<0.0001) and decreased LASEC (1.5+/-0.3; P<0.01); isoproterenol increased LAAEV (73.3+/-7.8 cm/s; P<0.0001) and decreased LASEC (0.3+/-0.2; P<0.01); and the postpacing pause increased LAAEV (68.3+/-3.8 cm/s; P<0.0001). In contrast, patients with long-duration AF demonstrated a significantly attenuated response of atrial mechanical function at each time point. With termination of AF, LAAEV decreased (19.1+/-2.6 to 8.2+/-1.0 cm/s; P=0.003) and LASEC increased (2.0+/-0.2 to 3.3+/-0.2; P<0.01); pacing increased LAAEV (18.4+/-2.7 cm/s; P<0.0001) and decreased LASEC (2.3+/-0.2; P<0.01); isoproterenol increased LAAEV (26.1+/-3.9 cm/s; P=NS to equivalent atrial rate) and decreased LASEC (1.0+/-0.3; P<0.01); and the postpacing pause increased LAAEV (27.2+/-2.4 cm/s; P=0.007). CONCLUSIONS: Atrial pacing at increased rates and isoproterenol can reverse atrial mechanical stunning associated with short duration AF. In contrast, long-duration AF is associated with an attenuated response to these maneuvers. These findings suggest a functional contractile apparatus in the mechanically remodeled atrium caused by AF; however, with longer durations of AF, additional factors may determine atrial mechanical function. PMID- 14557373 TI - Contrasting metabolic effects of antihypertensive agents. AB - Hypertension often coexists with hyperlipidemia, insulin resistance, and glucose intolerance, a comorbidity known as metabolic syndrome X. Different antihypertensives have mixed effects on these associated abnormalities. We compared three antihypertensives in the spontaneously hypertensive obese rat model of syndrome X. Moxonidine (4 mg/kg), an imidazoline and alpha2-adrenergic agonist, alpha-methyldopa (200 mg/kg), an alpha2-adrenergic agonist, or the vasodilator hydralazine (10 mg/kg) was given orally for 15 d. All three agents lowered blood pressure equally. Moxonidine significantly reduced fasting plasma insulin, glucagon, cholesterol, triglycerides, and free fatty acids (FFA) compared with untreated controls. In contrast, syndrome X markers were not affected by alpha-methyldopa treatment, and hydralazine reduced only glucagon and FFA. Relative to untreated controls, moxonidine improved glucose tolerance as shown by reduced glucose area under the curve (AUC) (13.6 +/- 2.4 versus 42.5 +/- 9.9 g x min/dl). Insulin AUC was increased (7.4 +/- 0.9 versus 3.9 +/- 1.8 microg x min/ml) as was the plasma C-peptide response to the glucose load. In contrast, alpha-methyldopa and hydralazine worsened glucose tolerance (68 +/- 26 and 110 +/ 21 g x min/ml, respectively) and significantly reduced insulin AUC (2.5 +/- 0.8 and -2.3 +/- 1.0 microg x min/ml, respectively) compared with controls. Moxonidine reduced but alpha-methyldopa and hydralazine elevated glucagon levels after the glucose load. Contrary to the "hemodynamic hypothesis" for the metabolic actions of antihypertensives, which predicts roughly equal benefits, only moxonidine had a positive impact on comorbidities. This unique action suggests a role for direct stimulation of imidazoline receptors. PMID- 14557374 TI - Generation and evaluation of a CYP2C9 heteroactivation pharmacophore. AB - Positive cooperativity (auto- and heteroactivation) of drug oxidation, a potential cause of drug interactions, is well established in vitro for cytochrome P450 (P450) 3A4 but to a much lesser extent for other drug-metabolizing P450 isoforms. Using a high throughput fluorescent-based CYP2C9 effector assay, we identified >30 heteroactivators from a set of 1504 structurally diverse compounds. Several potent heteroactivators of CYP2C9-mediated 7-methoxy-4 trifluoromethyl-coumarin metabolism are marketed drugs or endogenous compounds (amiodarone, niclosamide, liothyronine, meclofenemate, zafirlukast, estropipate, and dichlorphenamide, yielding 150% control reaction velocity at 0.04, 0.09, 0.5, 1, 1.2, 1.5, and 2.5 microM, respectively). Some heteroactivators are also known CYP2C9 substrates or inhibitors, suggesting potential multiple binding sites and substrate-dependent effects. v(150%), the concentration of effector giving 150% of control reaction velocity, was used as pharmacophore modeling parameter based on enzyme kinetic assumptions. The generated pharmacophore (training set: n = 36, v(150%) 0.04-150 microM) contains one hydrogen bond acceptor, one aromatic ring, and two hydrophobes. v(150%) values for 94% of the training set heteroactivators were predicted within 1 log unit for the residual (r [log observed v(150%)] versus [log predicted v(150%)] = 0.71; r2 0.50). The model also correctly identifies close to 70% of potent inhibitors (IC50 < 1 microM) as high-affinity CYP2C9 binders, suggesting that heteroactivators and inhibitors share some common structural CYP2C9 binding features, supporting the previously suggested hypothesis that CYP2C9 heteroactivators can bind within the active site. PMID- 14557375 TI - Repression of mitogen-activated protein kinase (MAPK) phosphatase-1 by anthracyclines contributes to their antiapoptotic activation of p44/42-MAPK. AB - Anthracyclines are commonly used chemotherapeutics, and in some models enhance p44/42-mitogen-activated protein kinase (MAPK) pathway signaling by effects on upstream kinases. To evaluate the impact of anthracyclines on p44/42-MAPK in breast cancer, A1N4-myc human mammary and BT-474 and MDA-MB-231 breast carcinoma cells were studied. Treatment with doxorubicin or epirubicin resulted in increased phospho-p44/42-MAPK levels in a time- and concentration-dependent manner. This was associated with p44/42 activation, as reflected by increased p90 ribosomal protein S6 kinase and Bad phosphorylation. Activation of p44/42 appeared to be antiapoptotic, since MAPK stimulation with epidermal growth factor or a dominant-positive p42 construct inhibited apoptosis. Modest activation of the upstream MAPK kinase MEK was noted under some conditions, but inhibition of MEK did not abolish p44/42 activation, suggesting a contribution from another mechanism. Anthracyclines were found to decrease expression of MAPK phosphatase-1 (MKP-1) both in vitro and in vivo. MKP-1 mRNA levels were decreased in anthracycline-treated cells, and transcription from the MKP-1 promoter was repressed. Inhibition of MKP-1 expression through the use of small interfering RNAs decreased the ability of anthracyclines to induce phospho-p44/42. Wild-type mouse embryo fibroblasts (MEFs) treated with doxorubicin showed increased phospho p44/42-MAPK levels, but MEFs from MKP-1 heterozygous and homozygous knockout mice had blunted p44/42 activation. These studies support the ability of anthracyclines to activate antiapoptotic p44/42-MAPK phosphorylation in breast cancer, and indicate that this occurs in part through the novel mechanism of repression of MKP-1 transcription. PMID- 14557376 TI - Differential effects of agonists on adenylyl cyclase superactivation mediated by the kappa opioid receptors: adenylyl cyclase superactivation is independent of agonist-induced phosphorylation, desensitization, internalization, and down regulation. AB - Prolonged activation of opioid receptors followed by agonist removal leads to adenylyl cyclase (AC) superactivation. In this study, we examined in CHO cells stably expressing the human or rat kappa opioid receptor (hkor or rkor) whether agonists had differential abilities to induce AC superactivation and whether the hkor and rkor exhibited differential AC superactivation. Pretreatment of the hkor with (trans)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl) cyclohexyl]benzeneacetamide methanesulfonate (U50,488H) induced AC superactivation in a time- and dose-dependent manner, reaching a plateau at 4 h and 0.1 microM. The extents of AC superactivation after a 4-h pretreatment of the hkor with saturating concentrations of agonists were in the order of the full agonists U50,488H, dynorphin A(1-17), (+/-)-ethylketocyclazocine, etorphine, and U69,593 > the high-efficacy partial agonist nalorphine > the low-efficacy partial agonists nalbuphine, morphine, and pentazocine. Interestingly, the full agonist levorphanol caused much lower AC superactivation than other full agonists and reduced the AC superactivation induced by U50,488H and dynorphin A(1-17) in a dose-dependent manner. The order of relative efficacies of agonists in causing AC superactivation mediated by the rkor was similar to that mediated by the hkor and the extents of AC superactivation were slightly lower. Because the rkor does not undergo U50,488H (1 microM)-induced phosphorylation, desensitization, internalization, and down-regulation in these cells, the degree of AC superactivation is independent of these processes. This is among the first reports to demonstrate that relative efficacies of agonists in causing AC superactivation generally correlated with those in activating G proteins and a full agonist reduced AC superactivation induced by another full agonist. PMID- 14557377 TI - Stereoselective and concentration-dependent polarized epithelial permeability of a series of phosphoramidate triester prodrugs of d4T: an in vitro study in Caco-2 and Madin-Darby canine kidney cell monolayers. AB - Nucleoside analogs are successful, widely used antiviral and anticancer therapeutics. Nucleotide prodrugs (i.e., pronucleotide) have increasingly been used to improve in vivo efficacy of nucleoside analogs. In this study, we evaluated the permeability of a series of phosphoramidate triester prodrugs of the anti-HIV drug 2',3'-didehydro-2',3'-dideoxythymidine across monolayers of Caco-2, Madin-Darby canine kidney (MDCKII) epithelial cell line, and its recombinant clone containing the human MDR1/P-gp gene (MDR1-MDCKII). Transport was studied in the apical-to-basolateral (A-B) and the basolateral-to-apical directions (B-A). The impact upon transport of differences in stereochemistry at the chiral phosphate center was evaluated. In the Caco-2 and MDCK models the A-B permeability was lower than expected based on the lipophilicity of the compounds, suggesting the involvement of a polarized efflux system and/or metabolic degradation in limiting the absorption of these ester-based prodrugs. Average permeability values through cell monolayers obtained in the A-B direction were lower than in the B-A direction. The inclusion of the P-glycoprotein (P-gp) inhibitor verapamil in the transport medium markedly increased the permeability in the A-B direction, whereas decreasing it in the opposite direction, suggesting an efflux mechanism mainly mediated by P-gp. Stereoselective permeability was significant for the most lipophilic compounds, where the diastereoisomer possessing the slower eluting time on a reverse-phase high-performance liquid chromatography column was transported through Caco-2 and MDCK monolayers at higher rate. PMID- 14557378 TI - Dynorphinergic mechanism mediating endomorphin-2-induced antianalgesia in the mouse spinal cord. AB - We have previously demonstrated that both endomorphin-1 (EM-1) and endomorphin-2 (EM-2) at high doses (1.75-35 nmol) given intrathecally (i.t.) or intracerebroventricularly produce antinociception by stimulation of mu-opioid receptors. Now, we report that EM-2 at small doses (0.05-1.75 nmol), which injected alone did not produce antinociception, produces anti-analgesia against opioid agonist-induced antinociception. The tail-flick (TF) response was used to test the antinociception in male CD-1 mice. Intrathecal pretreatment with EM-2 (0.02-1.75 nmol) 45 min before i.t. morphine (3.0 nmol) injection dose dependently attenuated morphine-induced TF inhibition. On the other hand, a similar dose of EM-1 (1.64 nmol) failed to produce any antianalgesic effect. The EM-2 (1.75 nmol)-produced anti-analgesia against morphine-induced TF inhibition was blocked by i.t. pretreatment with the mu-opioid antagonist naloxone or 3 methoxynaltrexone, but not delta-opioid receptor antagonist naltrindole, kappa opioid receptor antagonist nor-binaltorphimine, or N-methyl-d-aspartate (NMDA) receptor antagonist MK-801. The EM-2-induced antianalgesic effect against morphine-induced TF inhibition was blocked by i.t. pretreatment with antiserum against dynorphin A(1-17), but not beta-endorphin, [Met]-enkephalin, [Leu] enkephalin, or cholecystokinin antiserum (200 microg each). The i.t. EM-2 pretreatment also attenuated the TF inhibition induced by other mu-opioid agonists, [d-Ala2,N-Me-Phe4,Gly-ol5]-enkephalin, EM-1 and EM-2, delta-opioid agonist deltorphin II, and kappa-opioid agonist (trans)-3,4-dichloro-N-methyl-N [2-(1-pyrrolidinyl)-cyclohexyl]benzeneacetamide methane-sulfonate hydrate (U50,488H). It is concluded that EM-2 at subanalgesic doses presumably stimulates a subtype of mu-opioid receptor and subsequently induces the release of dynorphin A(1-17) to produce antianalgesic effects against mu-, delta-, or kappa-agonists induced antinociception. The EM-2-induced antianalgesia is not mediated by the release of [Met]-enkephalin, [Leu]-enkephalin, beta-endorphin, or cholecystokinin, nor does it involve kappa- or delta-opioid or NMDA receptors in the spinal cord. PMID- 14557379 TI - Induction of C-X-C chemokines, growth-related oncogene alpha expression, and epithelial cell-derived neutrophil-activating protein-78 by ML-1 (interleukin 17F) involves activation of Raf1-mitogen-activated protein kinase kinase extracellular signal-regulated kinase 1/2 pathway. AB - Neutrophil recruitment into the airway typifies pulmonary inflammation and is regulated through chemokine network, in which two C-X-C chemokines play a critical role. Airway epithelial cells and vein endothelial cells are major cell sources of chemokines. ML-1 (interleukin-17F) is a recently discovered cytokine and its function still remains elusive. In this report, we investigated the functional effect of ML-1 in the expression of growth-related oncogene (GRO)alpha and epithelial cell-derived neutrophil activating protein (ENA)-78. The results showed first that ML-1 induces, in time- and dose-dependent manners, the gene and protein expressions for both chemokines in normal human bronchial epithelial cells and human umbilical vein endothelial cells. Furthermore, selective mitogen activated protein kinase kinase (MEK) inhibitors 2'-amino-3'-methoxyflavone (PD98059), 1,4-diamino-2,3-dicyano-1,4-bis(o-aminophenylmercapto) butadiene (U0126), and Raf1 kinase inhibitor I partially inhibited Ml-1-induced GROalpha and ENA-78 production. In contrast, the combination of PD98059 and Raf1 kinase inhibitor I completely abrogated the chemokine production, whereas a protein kinase C inhibitor, 2-(1-(3-aminopropyl) indol-3-yl)-3-(1-methylindol-3-yl) maleimide, acetate (Ro-31-7549), and a phosphatidylinositol 3-kinase inhibitor, 2 (4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002), did not affect their production. Together, these data indicates a role for Raf1-MEK-extracellular signal-regulated kinase 1/2 pathway in ML-1 induced C-X-C chemokine expression, suggesting potential pharmacological targets for modulation. PMID- 14557380 TI - Opioid interactions in rhesus monkeys: effects of delta + mu and delta + kappa agonists on schedule-controlled responding and thermal nociception. AB - Agonists at delta, mu, and kappa opioid receptors produce interacting effects in rodents and nonhuman primates. To further evaluate the determinants of these interactions, this study examined the effects of mixtures of delta + mu and delta + kappa agonists in rhesus monkeys (n = 4-5) using two behavioral procedures, an assay of schedule-controlled responding for food reinforcement and an assay of thermal nociception. Results were analyzed using dose-addition analysis. In the assay of schedule-controlled responding, the delta agonist (+)-4-[(alphaR)-alpha ((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3-methoxy-benzyl]-N,N-diethyl benzamide (SNC80); the mu agonists methadone, fentanyl, morphine, and nalbuphine; and the kappa agonists (5alpha,7alpha,8beta)-(-)-N-methyl-N-(7-(1-pyrrolidinyl)-1 oxaspiro(4,5)dec-8-yl) benzeneacetamide (U69,593) and bremazocine all dose dependently decreased rates of food-maintained responding when administered alone. Fixed ratio mixtures of SNC80 + mu agonists produced additive or subadditive effects, whereas SNC80 + kappa agonist mixtures produced only additive effects. In the assay of thermal nociception, SNC80 produced no measurable effects when administered alone, whereas mu and kappa agonists produced dose-dependent antinociception. SNC80 + mu agonist mixtures produced superadditive effects manifested as leftward shifts in mu agonist dose-effect curves. This synergism was antagonized by the delta-selective antagonist naltrindole, suggesting that SNC80-induced enhancement of mu agonist antinociception was delta receptor-mediated. SNC80 did not enhance the antinociceptive effects of the highly selective kappa agonist U69,593, and it produced only a marginal enhancement of antinociception produced by the less selective kappa agonist bremazocine. These results suggest that delta agonists may selectively enhance the antinociceptive effects of mu agonists in rhesus monkeys. These results also confirm that opioid agonist interactions may depend on the receptor selectivity and relative doses of the agonists and on the experimental endpoint. PMID- 14557381 TI - Involvement of adenosine A1 and A2A receptors in the adenosinergic modulation of the discriminative-stimulus effects of cocaine and methamphetamine in rats. AB - Adenosine, by acting on adenosine A1 and A2A receptors, is known to antagonistically modulate dopaminergic neurotransmission. We have recently reported that nonselective adenosine receptor antagonists (caffeine and 3,7 dimethyl-1-propargylxanthine) can partially substitute for the discriminative stimulus effects of methamphetamine. In the present study, by using more selective compounds, we investigated the involvement of A1 and A2A receptors in the adenosinergic modulation of the discriminative-stimulus effects of both cocaine and methamphetamine. The effects of the A1 receptor agonist N6 cyclopentyladenosine (CPA; 0.01-0.1 mg/kg) and antagonist 8-cyclopentyl-1,3 dimethylxanthine (CPT; 1.3-23.7 mg/kg) and the A2A receptor agonist 2-p-(2 carboxyethyl)phenethylamino-5'-N-ethylcarboxamidoadenosine hydrochloride (CGS 21680; 0.03-0.18 mg/kg) and antagonist 3-(3-hydroxypropyl)-8-(3-methoxystyryl)-7 methyl-1-propargylxanthin phosphate disodium salt (MSX-3; 1-56 mg/kg) were evaluated in rats trained to discriminate either 1 mg/kg methamphetamine or 10 mg/kg cocaine from saline under a fixed-ratio 10 schedule of food presentation. The A1 and A2A receptor antagonists (CPT and MSX-3) both produced high levels of drug-lever selection when substituted for either methamphetamine or cocaine and significantly shifted dose-response curves of both psychostimulants to the left. Unexpectedly, the A2A receptor agonist CGS 21680 also produced drug-appropriate responding (although at lower levels) when substituted for the cocaine-training stimulus, and both CGS 21680 and the A1 receptor agonist CPA significantly shifted the cocaine dose-response curve to the left. In contrast, both agonists did not produce significant levels of drug-lever selection when substituted for the methamphetamine-training stimulus and failed to shift the methamphetamine dose-response curve. Therefore, adenosine A1 and A2A receptors appear to play important but differential roles in the modulation of the discriminative-stimulus effects of methamphetamine and cocaine. PMID- 14557382 TI - Rapid transcriptional suppression of rat cytochrome P450 genes by endotoxin treatment and its inhibition by curcumin. AB - Down-regulation of constitutive hepatic cytochrome P450 (P450) mRNAs by bacterial endotoxin (lipopolysaccharide, LPS) or other inflammatory stimuli has been documented extensively, but the contribution of transcriptional suppression to this effect is poorly understood. Here, we demonstrate that the rates of transcription of the CYP2C11, CYP3A2, and CYP2E1 genes are reduced to 20, 30, and 10% of control levels, respectively, in rat liver within 1 to 2 h of injection of LPS (1 mg/kg). The magnitude and rapidity of these effects indicate that transcriptional suppression is a primary reason for the decline in P450 mRNAs. Injection of curcumin significantly inhibited the rapid transcriptional suppression of CYP2E1, and blocked that of CYP3A2. These effects seemed to be independent of inhibition of nuclear factor-kappaB (NF-kappaB) activation by curcumin, because induction of known NF-kappaB-regulated genes was not attenuated. One hour after LPS injection, the DNA-binding activities of hepatocyte nuclear factor (HNF)1alpha, HNF3beta, and HNF4alpha were reduced to 73, 72, and 53%, respectively, of control values. The nuclear abundances of Sp1, liver-enriched transcriptional inhibitory protein (LIP), HNF1alpha, and HNF3beta were unchanged, whereas the abundance of HNF4alpha was reduced to 87% of control levels. We conclude that changes in Sp1 or LIP do not contribute significantly to the early suppression of P450 transcription in the acute phase rat liver. Although changes in DNA-binding activities of HNF1alpha, HNF3beta, and HNF4alpha are too small individually to explain the observed changes in P450 transcription, the role of each factor in concert with other factors remains to be determined. PMID- 14557383 TI - The pathogenic roles of tumor necrosis factor receptor p55 in acetaminophen induced liver injury in mice. AB - Acetaminophen (APAP) causes a massive production of intrahepatic tumor necrosis factor alpha (TNF-alpha). However, it still remains elusive regarding the roles of TNF-alpha in APAP-induced liver injury. Hence, we examined pathogenic roles of the TNF-alpha-TNF receptor with a molecular weight of 55 kDa (TNF-Rp55) axis in APAP-induced hepatotoxicity using TNF-Rp55-deficient [TNF-Rp55-knockout (KO)] mice. When wild-type (WT) BALB/c and TNF-Rp55-KO mice were intraperitoneally injected with a lethal dose of APAP (750 mg/kg), the mortality of TNF-Rp55-KO mice was marginally but significantly reduced compared with WT mice. Upon treatment with a nonlethal dose (600 mg/kg), WT mice exhibited an increase in serum transaminase levels. Histopathologically, centrilobular hepatic necrosis with leukocyte infiltration was evident at 10 and 24 h after APAP challenge. Moreover, mRNA expression of adhesion molecules, several chemokines, interferon gamma (IFN-gamma), and inducible nitric oxide synthase (iNOS) was enhanced in the liver. On the contrary, serum transaminase elevation and histopathological changes were attenuated in TNF-Rp55-KO mice injected with APAP (600 mg/kg). The gene expression of all molecules except for IFN-gamma and iNOS was significantly attenuated in TNF-Rp55-KO mice. Moreover, anti-TNF-alpha neutralizing antibodies alleviated liver injury when administered at 2 or 8 h after but not at 1 h before APAP challenge to WT mice. Collectively, the TNF-alpha-TNF-Rp55 axis has pathogenic roles in APAP-induced liver failure. PMID- 14557385 TI - Injury, sepsis, and the regulation of Toll-like receptor responses. AB - Although we tend to think that the immune system has evolved to protect the host from invading pathogens and to discriminate between self and nonself, there must also be an element of the immune system that has evolved to control the response to tissue injury. Moreover, these potential immune-regulatory pathways controlling the injury response have likely coevolved in concert with self and nonself discriminatory immune-regulatory networks with a similar level of complexity. From a clinical perspective, severe injury upsets normal immune function and can predispose the injured patient to developing life-threatening infectious complications. This remains a significant health care problem that has driven decades of basic and clinical research aimed at defining the functional effects of injury on the immune system. This review and update on our ongoing research efforts addressing the immunological response to injury will highlight some of the most recent advances in our understanding of the impact that severe injury has on the innate and adaptive immune system focusing on phenotypic changes in innate immune cell responses to Toll-like receptor stimulation. PMID- 14557384 TI - The chronic consequences of severe sepsis. AB - The early events of severe sepsis set in motion a cascade of events that significantly contributes to the morbidity and mortality observed during the first few days of this syndrome. Although sepsis is a deadly, acute disease, survivors also suffer long-term consequences. Clinical data underscore subsequent high mortality rates associated with patients who are long-term survivors of the acute septic episode. Within 1 year of surviving severe sepsis, there is a 26% predicted mortality rate, and many patients succumb to lung complications. In this review, we focus on the cellular and molecular mechanisms that dictate the longer-term sequela of sepsis and related lung injury. We have established a murine model of experimental sepsis [cecal ligation and puncture (CLP)], which results in an approximate 60% survival rate. Our studies have demonstrated that these survivors are susceptible to a fungal infection with 100% mortality when challenged 3 days or 15 days post-recovery from the initial CLP. This increased mortality correlates with changes in cytokines and Toll-like receptor expression and alterations in lung leukocyte populations. We hypothesize that the lung becomes predisposed to nosocomial infections for extended periods of time after severe sepsis via mechanisms that include alterations in inflammatory cytokines and an increase in immunomodulatory chemokines, such as monocyte chemoattractant protein-1 and C10. These mediators may alter the innate-immune response by affecting dendritic cells and macrophages, which could provide a mechanism for the immunosuppression observed following sepsis. PMID- 14557386 TI - Physiologic variations in granulocytic surface antigen expression: impact of age, gender, pregnancy, race, and stress. AB - There is a growing interest in the use of granulocytic surface markers for the diagnosis of some inherited and acquired disorders, such as Shwachman-Diamond syndrome and myelodysplastic syndromes. Understanding the impact of physiologic factors, such as age, gender, pregnancy, race, and stress on granulocytic surface markers is essential for appropriate interpretation of results. Some surface markers show marked variations at the very early and the very late stages in life. Fetal granulocytes tend to have a lower expression of CD11b, CD11c, CD18, and CD32. Term neonatal granulocytes are frequently associated with a lower expression of CD10, CD11b, CD13, CD33, and CD62L and a higher expression of CD55 and CD64. Elderly individuals have shown a higher expression of CD64. Pregnancy is associated with temporary changes in granulocytic surface markers, such as a lower expression of CD16 and a higher CD64, partially mimicking an inflammatory response. Stress also has an impact on some surface markers, particularly adhesion molecules, such as CD62L and CD54. These factors need to be taken in consideration for the optimal interpretation of granulocytic surface marker studies. PMID- 14557387 TI - Adoptive transfer of ex vivo immune-programmed NKT lymphocytes alleviates immune mediated colitis. AB - T lymphocyte-expressing natural killer (NK) cell markers (NKT cells) play a role in immune regulation. Our aim was to evaluate the in vivo effect of adoptive transfer of immune-programmed NKT cells. Colitis was induced in C57/B6 mice by 2,4,6-trinitrobenzenesulfonic acid. NKT, CD4, CD8 lymphocytes, and dendritic cells (DC) were prepared from spleens of naive mice, animals with colitis, and animals with colitis that were orally tolerized. Subsets of splenocytes, NKT, CD4, and CD8 and NKT+CD4, NKT+CD8, and NKT+DC lymphocytes were prepared. Assessment of the T helper cell type 1 (Th1)/Th2 cytokine secretion paradigm in vitro was performed before and following exposure to the antigen. Adoptive transfer of ex vivo immune-programmed lymphocytes from each group was performed into recipient mice, followed by colitis induction. Ex vivo exposure of NKT cells harvested from mice with colitis-to-colitis proteins [colitis-extracted proteins (CEP)] led to a Th2 cytokine shift. The interleukin (IL)-4/interferon-gamma (IFN gamma) ratio increased for NKT harvested from colitis-harboring mice following exposure to CEP. Adoptive transfer of NKT lymphocytes harvested from colitis harboring mice, which were ex vivo-educated, significantly alleviated experimental colitis in vivo. Intrahepatic NKT lymphocytes increased significantly in mice transplanted with NKT lymphocytes harvested from colitis harboring donor mice, which were ex vivo-exposed to CEP, similar to mice transplanted with NKT lymphocytes harvested from tolerized donors. Exposure of NKT cells to the disease-target antigen induced a significant increase in the IL 4/IFN-gamma cytokine ratio. Adoptive transfer of a relatively small number of immune-programmed NKT cells induced a systemic Th1 to Th2-immune shift and alleviated immune-mediated colitis. PMID- 14557388 TI - Inheritance, biochemical abnormalities, and clinical features of feline mucolipidosis II: the first animal model of human I-cell disease. AB - Mucolipidosis II (ML II), also called I-cell disease, is a unique lysosomal storage disease caused by deficient activity of the enzyme N-acetylglucosamine-1 phosphotransferase, which leads to a failure to internalize enzymes into lysosomes. We report on a colony of domestic shorthair cats with ML II that was established from a half-sibling male of an affected cat. Ten male and 9 female kittens out of 89 kittens in 26 litters born to clinically normal parents were affected; this is consistent with an autosomal recessive mode of inheritance. The activities of three lysosomal enzymes from affected kittens, compared to normal adult control cats, were high in serum (11-73 times normal) but low in cultured fibroblasts (9-56% of normal range) that contained inclusion bodies (I-cells), reflecting the unique enzyme defect in ML II. Serum lysosomal enzyme activities of adult obligate carriers were intermediate between normal and affected values. Clinical features in affected kittens were observed from birth and included failure to thrive, behavioral dullness, facial dysmorphia, and ataxia. Radiographic lesions included metaphyseal flaring, radial bowing, joint laxity, and vertebral fusion. In contrast to human ML II, diffuse retinal degeneration leading to blindness by 4 months of age was seen in affected kittens. All clinical signs were progressive and euthanasia or death invariably occurred within the first few days to 7 months of life, often due to upper respiratory disease or cardiac failure. The clinical and radiographic features, lysosomal enzyme activities, and mode of inheritance are homologous with ML II in humans. Feline ML II is currently the only animal model in which to study the pathogenesis of and therapeutic interventions for this unique storage disease. PMID- 14557389 TI - Microsatellite variation in Japanese and Asian horses and their phylogenetic relationship using a European horse outgroup. AB - The genetic relationships of seven Japanese and four mainland-Asian horse populations, as well as two European horse populations, were estimated using data for 20 microsatellite loci. Mongolian horses showed the highest average heterozygosities (0.75-0.77) in all populations. Phylogenetic analysis showed the existence of three distinct clusters supported by high bootstrap values: the European cluster (Anglo-Arab and thoroughbreds), the Hokkaido-Kiso cluster, and the Mongolian cluster. The relationships of these clusters were consistent with their geographical distributions. Basing our assumptions on the phylogenetic tree and the genetic variation of horse populations, we suggest that Japanese horses originated from Mongolian horses migrating through the Korean Peninsula. The genetic relationship of Japanese horses corresponded to their geographical distribution. Microsatellite polymorphism data were shown to be useful for estimating the genetic relationships between Japanese horses and Asian horses. PMID- 14557390 TI - Genetic diversity present within the near-complete mtDNA genome of 17 breeds of indigenous Chinese pigs. AB - The genetic diversity present within the near-complete mitochondrial genome (15,982 bp) was determined from 17 indigenous Chinese pig breeds and 3 European breeds. Animals were selected from 17 Chinese breeds that reflect the large phenotypic diversity of Chinese pigs and represent each of the six breed types, which are grouped based on morphological characteristics. Analysis of nucleotide diversity confirmed a high level of divergence between animals of European versus Asian origin; however, much more limited variation was observed between the 17 indigenous Chinese breeds. Each had a unique haplotype, but the lowest pairwise sequence divergence was only 0.01 +/- 0.01%, observed between the Tongcheng and Yushan Black. Comparison of control region sequence diversity revealed the 17 Chinese breeds contain a lower average pairwise distance (0.61 +/- 0.19%) than a group of European commercial breeds (0.91 +/- 0.21%). The dendrogram constructed from the near-complete mtDNA sequences showed the Chinese sequences loosely clustering into two groups. Although some correspondence with geographic origin was present, notable differences between the dendrogram and the traditional pig breed grouping system were observed. PMID- 14557391 TI - Mapping of 53 loci in American mink (Mustela vison). AB - Fifty-three genes were mapped in the American mink genome using polymerase chain reaction (PCR)-based analysis of a Chinese hamster-American mink somatic cell hybrid panel. Heterologous primers designed for cat gene mapping were used in this study. Forty-nine of these loci were localized into expected chromosome regions according to Zoo-FISH data, whereas four loci--ALPL, CDC20, ERF-2, and Fc(Mv)23617--were mapped out of expected conserved regions. PCR products amplified with primers corresponding to these four markers were partly sequenced and verified using BLAST. The results showed the homology to be more than 90% between mink and human or cat counterparts. At present, the gene map of American mink has expanded to 127 loci. PMID- 14557392 TI - On the number of genes controlling the grass stage in longleaf pine. AB - The grass stage is an inherent and distinctive developmental trait of longleaf pine (Pinus palustris), in which height growth in the first few years after germination is suppressed. In operational forestry practice the grass stage extends for two to several years and often plays a role in planting failures and decisions to plant alternative species. Interspecies hybrids involving loblolly (P. taeda) and slash (P. elliottii var. elliottii) pines have been investigated as a means to produce planting stock with improved early height growth and to develop backcross populations for advanced generation breeding. We have reevaluated data from several interspecies populations, with the objective of estimating the number of genes contributing to the difference in first-year height growth between longleaf and loblolly pines. Estimates based on means and variances of parental and interspecies hybrid and backcross families suggest a minimum of 4 to 10 genes with standard errors less than half the estimates. These results suggest that the grass stage has evolved through the accumulation of alleles at several loci, each with small effects on various components of first year height growth. Given the complexity of the grass-stage trait, tree breeders may need to combine genetic marker analysis with recurrent backcross breeding to efficiently develop longleaf pine planting stock for improved reforestation. PMID- 14557393 TI - Genetic structure, mating system, and long-distance gene flow in heart of palm (Euterpe edulis Mart.). AB - We report a detailed analysis of the population genetic structure, mating system, and gene flow of heart of palm (Euterpe edulis Mart.-Arecaceae) in central Brazil. This palm is considered a keystone species because it supplies fruits for birds and rodents all year and is intensively harvested for culinary purposes. Two populations of this palm tree were examined, using 18 microsatellite loci. The species displays a predominantly outcrossed mating system (tm = 0.94), with a probability of full sibship greater than 70% within open-pollinated families. The following estimates of interpopulation genetic variation were calculated and found significant: FIT = 0.17, FIS = 0.12, FST = 0.06, and RST = 0.07. This low but significant level of interpopulation genetic variation indicates high levels of gene flow. Two adult trees were identified as likely seed parents (P > 99.9%) of juveniles located at a distance of 22 km. Gene flow over such distances has not been reported before for tropical tree species. The establishment and management of in situ genetic reserves or ex situ conservation and breeding populations for E. edulis should contemplate the collection of several hundreds open-pollinated maternal families from relatively few distant populations to maximize the genetic sampling of a larger number of pollen parents. PMID- 14557394 TI - Microsatellite markers in avocado (Persea americana Mill.): genealogical relationships among cultivated avocado genotypes. AB - Twenty-five microsatellite markers uniquely differentiated 35 avocado cultivars and two wild relatives. Average heterozygosity was high (60.7%), ranging from 32% in P. steyermarkii to 84% in Fuerte and Bacon. In a subset of 15 cultivars, heterozygosity averaged 63.5% for microsatellites, compared to 41.8% for restriction fragment length polymorphisms (RFLPs). A neighbor-joining tree, according to average shared allele distances, consisted of three clusters likely corresponding to the botanical races of avocado and intermediate clusters uniting genotypes of presumably racially hybrid origin. Several results were at odds with existing botanical assignments that are sometimes rendered difficult by incomplete pedigree information, the complexity of the hybrid status (multiple backcrossing), or both. For example, cv. Harvest clustered with the Guatemalan race cultivars, yet it is derived from the Guatemalan x Mexican hybrid cv. Gwen. Persea schiedeana grouped with cv. Bacon. The rootstock G875 emerged as the most divergent genotype in our data set. Considerable diversity was found particularly among accessions from Guatemala, including G810 (West Indian race), G6 (Mexican race), G755A (hybrid Guatemalan x P. schiedeana), and G875 (probably not P. americana). Low bootstrap support, even upon exclusion of (known) hybrid genotypes from the data matrix, suggests the existence of ancient hybridization or that the botanical races originated more recently than previously thought. PMID- 14557395 TI - Genetic and molecular markers of the Queensland fruit fly, Bactrocera tryoni. AB - Twenty-six microsatellite markers, along with two restriction fragment length polymorphism (RFLP) markers and three morphological markers, have been mapped to five linkage groups, corresponding to the five autosomes of the Queensland fruit fly, Bactrocera tryoni. All these molecular and genetic markers were genotyped in three-generation pedigrees. Eight molecular markers were also localized to the salivary gland polytene chromosomes by in situ hybridization. This provides a substantial starting point for an integrated genetic and physical map of B. tryoni. PMID- 14557396 TI - Uniformly minimum variance unbiased estimation of gene diversity. AB - Gene diversity is an important measure of genetic variability in inbred populations. The survival of species in changing environments depends on, among other factors, the genetic variability of the population. In this communication, I have derived the uniformly minimum variance unbiased estimator of gene diversity. The proposed estimator of gene diversity does not assume that the inbreeding coefficient is known. I have also provided the approximate variance of this estimator according to Fisher's method. In addition, I have developed a numerical resampling-based method for obtaining variances and confidence intervals based on the maximum likelihood estimator and the uniformly minimum variance unbiased estimator. Efficiency in estimation of the gene diversity based on these two estimators is discussed. In accordance with the simulation results, I found that the uniformly minimum variance estimator developed in this report is more accurate for estimation of gene diversity than the maximum likelihood estimator. PMID- 14557397 TI - Molecular mapping of the male-sterile, female-sterile mutant gene (st8) in soybean. AB - Soybean male-sterile, female-sterile mutant genes have been identified by genetic and cytological studies. The St8 gene has been identified as an asynaptic mutation resulting in male and female sterility. This mutant gene was derived from a gene-tagging study using the soybean w4-mutable line. In this report we identified the genetic map position of st8 via restriction fragment length polymorphism (RFLP) and simple sequence repeat (SSR) markers. The St8 gene mutation was located between RFLP marker E107 and SSR markers Satt132, Sct_065, and Satt414 on molecular linkage group J and linked to each by 7.8 cM and 3.4 cM, respectively. PMID- 14557398 TI - DetSel 1.0: a computer program to detect markers responding to selection. AB - Estimating population parameters from polymorphism frequency data requires neutral genetic markers. Any departure from neutrality may invalidate the inferences drawn from such analyses. We recently discussed the possibility of identifying markers that show deviation from neutral expectations in pairwise comparisons of diverging populations. We are now releasing a user-friendly software package that implements all the necessary steps to identify the signature of selection among molecular markers in a set of polymorphism data. This software can be downloaded free of charge at http://www.univ montp2.fr/~genetix/detsel/detsel.html. PMID- 14557401 TI - Sodium and chloride concentrations, pH, and depth of airway surface liquid in distal airways. AB - The composition and depth of the airway surface liquid (ASL) are key parameters in airway physiology that are thought to be important in the pathophysiology of cystic fibrosis and other diseases of the airways. We reported novel fluorescent indicator and microscopy methods to measure [Na+], [Cl-], pH, and depth of the ASL in large airways (Jayaraman, S., Y. Song, L. Vetrivel, L. Shankar, and A.S. Verkman. 2001. J. Clin. Invest. 107:317-324.). Here we report a stripped-lung preparation to measure ASL composition and depth in small distal airways. Distal ASL was stained with ion- or pH-sensitive fluorescent indicators by infusion into mouse trachea of a perfluorocarbon suspension of the indicator. After stripping the pleura and limited microdissection of the lung parenchyma, airways were exposed for measurement of ASL [Na+], [Cl-], and pH by ratio imaging microscopy, and depth by confocal microscopy. The stripped-lung preparation was validated in stability and tissue viability studies. ASL [Na+] was 122 +/- 2 mM, [Cl-] was 123 +/- 4 mM and pH was 7.28 +/- 0.07, and not dependent on airway size (<100- to >250-mum diameter), ENaC inhibition by amiloride, or CFTR inhibition by the thiazolidinone CFTRinh-172. ASL depth was 8-35 mum depending on airway size, substantially less than that in mouse trachea of approximately 55 mum, and not altered significantly by amiloride. These results establish a novel lung preparation and fluorescence approach to study distal airway physiology and provide the first data on the composition and depth of distal ASL. PMID- 14557402 TI - The role of loop 5 in acetylcholine receptor channel gating. AB - Nicotinic acetylcholine receptor channel (AChR) gating is an organized sequence of molecular motions that couples a change in the affinity for ligands at the two transmitter binding sites with a change in the ionic conductance of the pore. Loop 5 (L5) is a nine-residue segment (mouse alpha-subunit 92-100) that links the beta4 and beta5 strands of the extracellular domain and that (in the alpha subunit) contains binding segment A. Based on the structure of the acetylcholine binding protein, we speculate that in AChRs L5 projects from the transmitter binding site toward the membrane along a subunit interface. We used single channel kinetics to quantify the effects of mutations to alphaD97 and other L5 residues with respect to agonist binding (to both open and closed AChRs), channel gating (for both unliganded and fully-liganded AChRs), and desensitization. Most alphaD97 mutations increase gating (up to 168-fold) but have little or no effect on ligand binding or desensitization. Rate-equilibrium free energy relationship analysis indicates that alphaD97 moves early in the gating reaction, in synchrony with the movement of the transmitter binding site (Phi = 0.93, which implies an open-like character at the transition state). alphaD97 mutations in the two alpha subunits have unequal energetic consequences for gating, but their contributions are independent. We conclude that the key, underlying functional consequence of alphaD97 perturbations is to increase the unliganded gating equilibrium constant. L5 emerges as an important and early link in the AChR gating reaction which, in the absence of agonist, serves to increase the relative stability of the closed conformation of the protein. PMID- 14557403 TI - Constitutive activation of the Shaker Kv channel. AB - In different types of K+ channels the primary activation gate is thought to reside near the intracellular entrance to the ion conduction pore. In the Shaker Kv channel the gate is closed at negative membrane voltages, but can be opened with membrane depolarization. In a previous study of the S6 activation gate in Shaker (Hackos, D.H., T.H. Chang, and K.J. Swartz. 2002. J. Gen. Physiol. 119:521 532.), we found that mutation of Pro 475 to Asp results in a channel that displays a large macroscopic conductance at negative membrane voltages, with only small increases in conductance with membrane depolarization. In the present study we explore the mechanism underlying this constitutively conducting phenotype using both macroscopic and single-channel recordings, and probes that interact with the voltage sensors or the intracellular entrance to the ion conduction pore. Our results suggest that constitutive conduction results from a dramatic perturbation of the closed-open equilibrium, enabling opening of the activation gate without voltage-sensor activation. This mechanism is discussed in the context of allosteric models for activation of Kv channels and what is known about the structure of this critical region in K+ channels. PMID- 14557410 TI - Abstracts of the International Conference on Water-Saving Agriculture and Sustainable Use of Water and Land Resources (ICWSAWLR). Yangling, Shaanixi, P. R. China. October 26-29, 2003. PMID- 14557404 TI - Movements near the gate of a hyperpolarization-activated cation channel. AB - Hyperpolarization-activated cation (HCN) channels regulate pacemaking activity in cardiac cells and neurons. Like the related depolarization-activated K+ channels (Kv channels), HCN channels use an intracellular activation gate to regulate access to an inner cavity, lined by the S6 transmembrane regions, which leads to the selectivity filter near the extracellular surface. Here we describe two types of metal interactions with substituted cysteines in the S6, which alter the voltage-controlled movements of the gate. At one position (L466), substitution of cysteine in all four subunits allows Cd2+ ions at nanomolar concentration to stabilize the open state (a "lock-open" effect). This effect depends on native histidines at a nearby position (H462); the lock-open effect can be abolished by changing the histidines to tyrosines, or enhanced by changing them to cysteines. Unlike a similar effect in Kv channels, this effect depends on a Cd2+ bridge between 462 and 466 in the same subunit. Cysteine substitution at another position (Q468) produces two effects of Cd2+: both a lock-open effect and a dramatic slowing of channel activation-a "lock-closed" effect. The two effects can be separated, because the lock-open effect depends on the histidine at position 462. The novel lock-closed effect results from stabilization of the closed state by the binding of up to four Cd2+ ions. During the opening conformational change, the S6 apparently moves from one position in which the 468C cysteines can bind four Cd2+ ions, possibly as a cluster of cysteines and cadmium ions near the central axis of the pore, to another position (or flexible range of positions) where either 466C or 468C can bind Cd2+ in association with the histidine at 462. PMID- 14557411 TI - The adaptor protein AP-3 is required for CD1d-mediated antigen presentation of glycosphingolipids and development of Valpha14i NKT cells. AB - Relatively little is known about the pathway leading to the presentation of glycolipids by CD1 molecules. Here we show that the adaptor protein complex 3 (AP 3) is required for the efficient presentation of glycolipid antigens that require internalization and processing. AP-3 interacts with mouse CD1d, and cells from mice deficient for AP-3 have increased cell surface levels of CD1d and decreased expression in late endosomes. Spleen cells from AP-3-deficient mice have a reduced ability to present glycolipids to natural killer T (NKT) cells. Furthermore, AP-3-deficient mice have a significantly reduced NKT cell population, although this is not caused by self-tolerance that might result from increased CD1d surface levels. These data suggest that the generation of the endogenous ligand that selects NKT cells may also be AP-3 dependent. However, the function of MHC class II-reactive CD4+ T lymphocytes is not altered by AP-3 deficiency. Consistent with this divergence from the class II pathway, NKT cell development and antigen presentation by CD1d are not reduced by invariant chain deficiency. These data demonstrate that the AP-3 requirement is a particular attribute of the CD1d pathway in mice and that, although MHC class II molecules and CD1d are both found in late endosomes or lysosomes, different pathways mediate their intracellular trafficking. PMID- 14557412 TI - Targeted inactivation of the IL-4 receptor alpha chain I4R motif promotes allergic airway inflammation. AB - The insulin/interleukin-4 (IL-4) receptor (I4R) motif mediates the association of insulin receptor substrate (IRS)-2 with the interleukin-4 (IL-4)Ralpha chain and transduces mitogenic signals in response to IL-4. Its physiological functions were analyzed in mice with a germline point mutation that changed the motif's effector tyrosine residue into phenylalanine (Y500F). The Y500F mutation abrogated IRS-2 phosphorylation and impaired IL-4-induced CD4+ T lymphocyte proliferation but left unperturbed Stat6 activation, up-regulation of IL-4 responsive gene products, and Th cell differentiation under Th2 polarizing conditions. However, in vivo the Y500F mutation was associated with increased allergen-induced IgE production, airway responsiveness, tissue eosinophilia, and mucus production. These results define an important role for the I4R motif in regulating allergic inflammation. PMID- 14557413 TI - Normal induction but attenuated progression of germinal center responses in BAFF and BAFF-R signaling-deficient mice. AB - The factors regulating germinal center (GC) B cell fate are poorly understood. Recent studies have defined a crucial role for the B cell-activating factor belonging to TNF family (BAFF; also called BLyS) in promoting primary B cell survival and development. A role for this cytokine in antigen-driven B cell responses has been suggested but current data in this regard are limited. A BAFF receptor expressed by B cells (BAFF-R/BR3) is defective in A/WySnJ mice which exhibit a phenotype similar to BAFF-deficient (BAFF-/-) animals. Here, we show that although GC responses can be efficiently induced in both A/WySnJ and BAFF-/- mice, these responses are not sustained. In BAFF-/- mice, this response is rapidly attenuated and accompanied by perturbed follicular dendritic cell development and immune complex trapping. In contrast, analysis of the A/WySnJ GC response revealed a B cell autonomous proliferative defect associated with reduced or undetectable Ki67 nuclear proliferation antigen expression by GC B cells at all stages of the response. These data demonstrate a multifaceted role for the BAFF pathway in regulating GC progression. PMID- 14557414 TI - Prevention of lipopolysaccharide-induced microangiopathy by gp49B1: evidence for an important role for gp49B1 expression on neutrophils. AB - gp49B1 is expressed on mast cells and inhibits immunoglobulin E-dependent activation and inflammation in vivo. We now show that gp49B1 is expressed on neutrophils and prevents neutrophil-dependent vascular injury in response to lipopolysaccharide (LPS). The intradermal (i.d.) injection of LPS into gp49B1 null (gp49B-/-) but not gp49B1-sufficient (gp49B+/+) mice elicited macroscopic hemorrhages by 24 h, which were preceded on microscopic analyses by significantly more intravascular thrombi (consisting of neutrophils, platelets, and fibrin) that occluded venules and by more tissue neutrophils than in gp49B+/+ mice. However, there were no differences in the number of intact (nondegranulating) mast cells or the tissue levels of mediators that promote neutrophil recruitment. Hemorrhage was prevented by depleting neutrophils, blocking beta2 integrin intercellular adhesion molecule 1 interactions, or inhibiting coagulation. These characteristics indicate that gp49B-/- mice are exquisitely sensitive to a local Shwartzman reaction (LSR) after a single i.d. injection of LPS, whereas in the classic LSR, a second exposure is required for increased beta2 integrin function, intravascular neutrophil aggregation, formation of occlusive thrombi, and hemorrhage. Moreover, LPS increased gp49B1 expression on neutrophils in vivo. The results suggest that gp49B1 suppresses the LPS-induced increase in intravascular neutrophil adhesion, thereby providing critical innate protection against a pathologic response to a bacterial component. PMID- 14557416 TI - Clinical review 163: Cardiovascular endocrinology: Low-density lipoprotein size and cardiovascular disease: a reappraisal. PMID- 14557415 TI - Transforming growth factor (TGF)-beta1-producing regulatory T cells induce Smad mediated interleukin 10 secretion that facilitates coordinated immunoregulatory activity and amelioration of TGF-beta1-mediated fibrosis. AB - Interleukin (IL)-10 and transforming growth factor (TGF)-beta1 are suppressor cytokines that frequently occur together during a regulatory T cell response. Here we used a one gene doxycycline (Dox)-inducible plasmid encoding TGF-beta1 to analyze this association and test its utility. In initial studies, we showed that intranasal administration of this plasmid (along with Dox) led to the appearance of TGF-beta1-producing cells (in spleen and lamina propria) and the almost concomitant appearance of IL-10-producing cells. Moreover, we showed that these cells exert Dox-regulated suppression of the T helper cell (Th)1-mediated inflammation in trinitrobenzene sulfonic acid colitis. In subsequent in vitro studies using retroviral TGF-beta1 expression, we established that IL-10 production by Th1 cells occurs after exposure to TGF-beta1 from either an endogenous or exogenous source. In addition, using a self-inactivating retrovirus luciferase reporter construct we showed that TGF-beta1 induces Smad4, which then binds to and activates the IL-10 promoter. Furthermore, intranasal TGF-beta1 plasmid administration ameliorates bleomycin-induced fibrosis in wild-type but not IL-10-deficient mice, strongly suggesting that the amelioration is IL-10 dependent and that IL-10 protects mice from TGF-beta1-mediated fibrosis. Taken together, these findings suggest that the induction of IL-10 by TGF-beta1 is not fortuitous, but instead fulfills important requirements of TGF-beta1 function after its secretion by regulatory T cells. PMID- 14557417 TI - Clinical review 164: The laboratory diagnosis of adrenal pheochromocytoma: the Mayo Clinic experience. PMID- 14557418 TI - In search of the impossible dream? Thyroid hormone replacement therapy that treats all symptoms in all hypothyroid patients. PMID- 14557419 TI - Combined thyroxine/liothyronine treatment does not improve well-being, quality of life, or cognitive function compared to thyroxine alone: a randomized controlled trial in patients with primary hypothyroidism. AB - T(4) is standard treatment for hypothyroidism. A recent study reported that combined T(4)/liothyronine (T(3)) treatment improved well-being and cognitive function compared with T(4) alone. We conducted a double-blind, randomized, controlled trial with a crossover design in 110 patients (101 completers) with primary hypothyroidism in which liothyronine 10 micro g was substituted for 50 micro g of the patients' usual T(4) dose. No significant (P < 0.05) difference between T(4) and combined T(4)/T(3) treatment was demonstrated on cognitive function, quality of life scores, Thyroid Symptom Questionnaire scores, subjective satisfaction with treatment, or eight of 10 visual analog scales assessing symptoms. For the General Health Questionnaire-28 and visual analog scales assessing anxiety and nausea, scores were significantly (P < 0.05) worse for combined treatment than for T(4) alone. Serum TSH was lower during T(4) treatment than during combined T(4)/T(3) treatment (mean +/- SEM, 1.5 +/- 0.2 vs. 3.1 +/- 0.2 mU/liter; P < 0.001), a potentially confounding factor; however, subgroup analysis of subjects with comparable serum TSH concentrations during each treatment showed no benefit from combined treatment compared with T(4) alone. We conclude that in the doses used in this study, combined T(4)/T(3) treatment does not improve well-being, cognitive function, or quality of life compared with T(4) alone. PMID- 14557420 TI - Does a combination regimen of thyroxine (T4) and 3,5,3'-triiodothyronine improve depressive symptoms better than T4 alone in patients with hypothyroidism? Results of a double-blind, randomized, controlled trial. AB - Some hypothyroid patients receiving levothyroxine replacement therapy complain of depressive symptoms despite normal TSH measurements. It is not known whether adding T(3) can reverse such symptoms. We randomized 40 individuals with depressive symptoms who were taking a stable dose of levothyroxine for treatment of hypothyroidism (excluding those who underwent thyroidectomy or radioactive iodine ablation of the thyroid) to receive T(4) plus placebo or the combination of T(4) plus T(3) in a double-blind manner for 15 wk. Participants receiving combination therapy had their prestudy dose of T(4) dropped by 50%, and T(3) was started at a dose of 12.5 micro g, twice daily. T(4) and T(3) doses were adjusted to keep goal TSH concentrations within the normal range. Compared with the group taking T(4) alone, the group taking both T(4) plus T(3) did not report any improvement in self-rated mood and well-being scores that included all subscales of the Symptom Check-List-90, the Comprehensive Epidemiological Screen for Depression, and the Multiple Outcome Study (P > 0.05 for all indexes). In conclusion, the current data do not support the routine use of combined T(3) and T(4) therapy in hypothyroid patients with depressive symptoms. PMID- 14557421 TI - Insulin action and distribution of tissue blood flow. PMID- 14557422 TI - Direct measurements of the permeability surface area for insulin and glucose in human skeletal muscle. AB - To elucidate mechanisms regulating capillary transport of insulin and glucose, we directly calculated the permeability surface (PS) area product for glucose and insulin in muscle. Intramuscular microdialysis in combination with the forearm model and blood flow measurements was performed in healthy males, studied during an oral glucose tolerance test or during a one-step or two-step euglycemic hyperinsulinemic clamp. PS for glucose increased significantly from 0.29 +/- 0.1 to 0.64 +/- 0.2 ml/min.100 g after oral glucose tolerance test, and glucose uptake increased from 1.2 +/- 0.4 to 2.6 +/- 0.6 micro mol/min.100 g (P < 0.05). During one-step hyperinsulinemic clamp (plasma insulin, 1.962 pmol/liter), PS for glucose increased from 0.2 +/- 0.1 to 2.3 +/- 0.9 ml/min.100 g (P < 0.05), and glucose uptake increased from 0.6 +/- 0.2 to 5.0 +/- 1.4 micro mol/min.100 g (P < 0.05). During the two-step clamp (plasma insulin, 1380 +/- 408 and 3846 +/- 348 pmol/liter), the arterial-interstitial difference and PS for insulin were constant. The PS for glucose tended to increase (P = not significant), whereas skeletal muscle blood flow increased from 4.4 +/- 0.7 to 6.2 +/- 0.8 ml/min.100 ml (P < 0.05). The present data show that PS for glucose is markedly increased by oral glucose, whereas a further vasodilation exerted by high insulin concentrations may not be physiologically relevant for capillary delivery of either glucose or insulin in resting muscle. PMID- 14557423 TI - Cushing's syndrome due to medullary thyroid carcinoma: diagnosis by proopiomelanocortin messenger ribonucleic acid in situ hybridization. AB - Medullary thyroid carcinoma (MTC) rarely causes ectopic ACTH syndrome. We describe a 38-yr-old man with renal stones who had a 5-cm MTC removed in 1992. He was RET-protooncogene positive (codon 618). Serum calcitonin was 1597 pg/ml postoperatively. In 1996 he had rib fractures, bruising, weakness, and three to four stools per day. Laboratory studies revealed an elevated 24-h urine-free cortisol (780 micro g/d), epinephrine (66 micro g/d), and calcium (558 mg/d). Baseline serum cortisol was 23.9 micro g/dl and decreased to 12.9 and 4.5 micro g/dl after 2 mg and 8 mg dexamethasone suppression, respectively. Plasma ACTH was 170 pg/ml and decreased to 75 and 24 pg/ml after dexamethasone. Bone density t score was -4.3 (trochanter). Computed tomography scans showed multiple cervical nodes and 2-cm right adrenal nodule. Magnetic resonance imaging (MRI) scan showed a prominent, homogeneous pituitary; the adrenal MRI scan was not typical for a pheochromocytoma. Serum CRH was less than 6.6 pg/ml. Bilateral adrenalectomy revealed two adjacent right adrenal pheochromocytomas and corrected the elevated urine cortisol (30 micro g/d), epinephrine (0 micro g/d), and calcium (281 mg/d) but not plasma ACTH (125 pg/ml). Neck dissection reduced calcitonin by 96% (5300 to 120 pg/ml) and ACTH by 91% (125 to 11 pg/ml). Carcinoembryonic antigen was reduced from 32.0 to 2.3 ng/ml. Immunohistochemical stain was negative for ACTH in the MTC-positive lymph nodes and the pheochromocytoma. Proopiomelanocortin mRNA by in situ hybridization was positive in the MTC but not in the pheochromocytoma. A repeat pituitary MRI scan was normal. The differential diagnosis of ACTH-dependent Cushing's syndrome in this case included pituitary disease or ectopic ACTH, either from medullary thyroid carcinoma or pheochromocytoma. ACTH stains were unrevealing, but proopiomelanocortin mRNA in situ hybridization in MTC tissue and plasma ACTH response to neck dissection confirmed MTC as the source of ectopic ACTH. PMID- 14557424 TI - Effect of pamidronate treatment in children with polyostotic fibrous dysplasia of bone. AB - Intravenous infusions with the bisphosphonate compound pamidronate decrease bone pain and reportedly can lead to refilling of dysplastic lesions in adults with fibrous dysplasia (FD) of bone. Here we describe the effects of this treatment approach in 18 children and adolescents (age at start of therapy, 6.2-17.5 yr; eight girls) with polyostotic FD, who received pamidronate for 1.2-9.1 yr (median, 3.8 yr). Treatment cycles with pamidronate (1-1.5 mg/kg.d on 3 consecutive days) were given every 4 months. Levels of serum alkaline phosphatase and urinary collagen type I N-telopeptide were elevated at baseline and decreased continuously during the first 3 yr of therapy. There was no radiographic evidence of filling of lytic lesions or thickening of the bone cortex surrounding the lesions in any patient. Histomorphometric results in dysplastic bone tissue of patients receiving pamidronate (n = 7; time of therapy, 1.4-4.8 yr) were similar to those of patients without medical therapy (n = 9). No serious side effects were noted. In conclusion, pamidronate therapy appears to be safe in children and adolescents with polyostotic FD. However, we found no clear evidence that pamidronate has an effect on dysplastic lesions in such patients. PMID- 14557425 TI - Reversible metaphyseal dysplasia, a novel bone phenotype, in two unrelated children with autoimmunepolyendocrinopathy-candidiasis-ectodermal dystrophy: clinical and molecular studies. AB - We report the association of an undescribed, reversible metaphyseal dysplasia (RMD) with autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) in two patients, one homozygous and one heterozygous for a 13-bp deletion in exon 8 of the autoimmune regulator (AIRE) gene. One patient also had a novel deletion in exon 6, resulting in a frameshift mutation and introduction of a STOP codon in exon 10. Their APECED phenotypes differed, but both patients developed progressive skeletal deformities and growth failure from early childhood. Radiological examination suggested a generalized abnormality of endochondral ossification, with irregular, flared, radioopaque regions in the metaphyses, subjacent to the growth plates. Histopathology in patient 1 showed islands of calcified cartilage within bone, consistent with impaired coupling of cartilage resorption with vascular invasion and ossification. Despite discordance for puberty, both patients experienced radiological resolution of their bone disease in their mid-teens, with improvement in histopathology in patient 1. RMD may constitute a rare phenotypic variation of APECED, possibly resulting from autoimmunity directed against skeletal proteins. We also demonstrated AIRE expression in chondrocytes derived from human fetal growth plates, primary culture of human chondrocytes, and two chondrosarcoma cell lines, suggesting a potential role for abnormal AIRE expression in the development of RMD. PMID- 14557426 TI - Are adult patients with Laron syndrome osteopenic? A comparison between dual energy X-ray absorptiometry and volumetric bone densities. AB - Severe short stature resulting from a deficiency in IGF-I is a prominent feature of Laron syndrome (LS). Although low bone mineral density (BMD) has been noted in LS patients examined by dual energy x-ray absorptiometry (DEXA), this technique does not take volume into account and may therefore underestimate the true bone density in patients with small bones. The aim of the present study was to evaluate the BMD yielded by DEXA in our LS patients using estimated volumetric values. Volumetric density was calculated with the following formulas: bone mineral apparent density (BMAD) = bone mineral content (BMC)/(area)(3/2) for the lumbar spine and BMAD = BMC/area(2) for the femoral neck. The study sample included 12 patients (mean age, 43.9 yr; mean height, 123.7 cm). Findings were compared with 10 osteopenic subjects without developmental abnormalities (mean age, 56 yr; mean height, 164.8 cm) and 10 healthy control subjects matched for sex and age to the LS patients (mean height, 165.5 cm). BMAD in the LS group was 0.201 +/- 0.02 g/cm(3) at the lumbar spine and 0.201 +/- 0.04 g/cm(3) at the femoral neck; corresponding values for the osteopenic group were 0.130 +/- 0.01 and 0.140 +/- 0.01 g/cm(3), and for the controls, 0.178 +/- 0.03 and 0.192 +/- 0.02 g/cm(3). Although areal BMD was significantly lower in the LS and osteopenic subjects compared with controls (P < 0.02) at both the lumbar spine and femoral neck, BMAD was low (P < 0.01) in the osteopenic group only. In conclusion, DEXA does not seem to be a reliable measure of osteoporosis in patients with LS. PMID- 14557427 TI - Association of morphological characteristics with precocious puberty and/or gelastic seizures in hypothalamic hamartoma. AB - The pathogenesis of central precocious puberty (PP) and/or gelastic seizures due to a hypothalamic hamartoma (HH) is still under debate. We evaluated the association of clinical symptoms with morphology and localization of the HH in 34 patients. The majority (86.4%) of HHs in patients with isolated PP (n = 22; 68.2% females) revealed a parahypothalamic position without affecting the third ventricle (91%). Half of them were pedunculated, and 40.9% showed a diameter less than 10 mm. In contrast, 11 of 12 patients with seizures, eight of whom were male, presented with a sessile intrahypothalamic hamartoma, 10 of which distorted the third ventricle. Logistic regression analysis revealed an increased relative risk (RR) for epilepsy in males (RR, 4.3; 95% confidence interval, 0.96-19). However, combination of the risk factor gender with intrahypothalamic position (RR, 19; 1.3-285) and distortion of the third ventricle (RR, 10; 0.6-164) reduced the risk associated with male gender to 1.1. The position of a HH and involvement of the third ventricle are likely to be more predictive for clinical characteristics than size and shape. Male gender was associated with an intrahypothalamic HH and epilepsy, suggesting a sexually dimorphic developmental pattern of this heterotopic mass. PMID- 14557428 TI - Is glycosylated hemoglobin A1c a surrogate for metabolic syndrome in nondiabetic, first-degree relatives of African-American patients with type 2 diabetes? AB - Glycosylated hemoglobin (Hb)A1c provides a practical assessment of long-term glycemic control in patients with diabetes. However, whether HbA1c has any clinical significance in metabolic syndrome (MS) in nondiabetic subjects remains debatable. Therefore, we examined the impact of different levels of HbA1c on insulin sensitivity (Si), non-insulin-dependent glucose disposal, and blood pressure (BP), as well as lipids and lipoproteins in nondiabetic, first-degree relatives of African-American patients with type 2 diabetes. The study consisted of 219 nondiabetic, first-degree relatives (offspring and siblings) of African American patients with type 2 diabetes. To examine the metabolic impact of HbA1c in our population, HbA1c was divided into tertiles (normal range, 3.3-6.4%). The mean HbA1c was 4.7% (range, 3.3-4.8%, n = 74) for tertile 1, 5.4% (range, 4.9 5.6%, n = 73) for tertile 2, and 5.8% (range, 5.7-6.4%, n = 72) for tertile 3. Si and glucose effectiveness (Sg) were determined by the Bergman's minimal model method. Homeostasis model assessment (HOMA)-insulin resistance and HOMA-beta-cell function were also estimated. BP, body compositional variables, and body fat distribution, as well as fasting serum lipid and lipoprotein concentrations, were determined in each subject. The mean age, body weight, body mass index, waist and hip circumference, and systolic and diastolic BPs were significantly (P < 0.02 0.001) greater in the subjects in tertile 3 than those in tertiles 1 and 2. The mean fasting serum glucose was significantly (P < 0.01) higher in tertile 3 (95.5 +/- 3.2 mg/dl) than in tertile 2 (83.0 +/- 2.7 mg/dl) and tertile 1 (78.8 +/- 1.5 mg/dl). Mean fasting serum insulin and c-peptide levels tended to be higher in tertile 3 subjects than in those in tertiles 1 and 2, but the mean differences did not reach statistical significance. The mean Si was significantly (P < 0.001) lower in the subjects in tertile 3 [1.66 +/- 0.2019 x 10(-4).min(-1)( micro U/ml)(-1)], when compared with those in tertile 1 [2.27 +/- 0.20 19 x 10(-4).min( 1)( micro U/ml)(-1)] and tertile 2 [2.61 +/- 0.19 x 10(-4).min(-1)( micro U/ml)( 1)]. The mean Sg was significantly (P < 0.02) lower in tertile 3 (1.95 +/- 0.12 x 10(-2).min(-1)), when compared with those of tertile 1 (2.27 +/- 0.10 x 10( 2).min(-1)) and tertile 2 (2.29 +/- 0.11 x 10(-2).min(-1)). In addition, the (HOMA)-insulin resistance was significantly (P < 0.01) higher in tertile 3 (3.62 +/- 0.26) than in tertile 1 (2.6 +/- 0.21) and tertile 2 (2.55 +/- 0.31) HbA1c. In contrast, HOMA-beta-cell function, was not different among tertiles 1, 2, and 3. Mean fasting serum triglycerides, cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol levels also were not significantly different in subjects in tertile 3, when compared with those in tertiles 1 and 2. In summary, the present study demonstrates that the upper tertile HbA1c level (tertile 3) reflects some components of MS in the nondiabetic, obese, first-degree relatives of African-Americans who are genetically predisposed to type 2 diabetes. The metabolic abnormalities in the upper tertile 3 subjects included a reduced insulin action (Si) and reduced Sg, as well as elevated systolic and diastolic BPs, but not beta-cell secretion and lipids and lipoproteins. We conclude that the upper tertile of HbA1c should be considered as a major surrogate of MS in high-risk African-Americans who are genetically predisposed to type 2 diabetes. PMID- 14557429 TI - The hypoparathyroidism of autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy protective effect of male sex. AB - In autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy, hypoparathyroidism (HP) is the most common endocrine component. It occurs in most (but not all) patients. Determinants of its occurrence are unknown, and there is no proof for its autoimmune nature. Recently, the Ca(2+)-sensing receptor (CaSR) was reported to be an autoantigen in HP. With our group of 90 patients, we aimed at identifying the determinants and pathomechanism of HP. For the determinants, we evaluated gender and the HLA class II. For the pathomechanism, we searched for parathyroid autoantibodies, including antibodies against CaSR and PTH. Also, we studied whether AIRE is expressed in the human parathyroid, because its absence could be a pathogenetic factor. We found a clear gender linkage with lower and later incidence in males. Of the 14 patients who had escaped HP, 13 were males. This was associated with adrenal failure, which was the first or only endocrinopathy in 47% of males vs. 7% of females. In contrast, we found no linkage to the HLA class II. By immunofluorescence, 19% of the patients had antibodies to parathyroid epithelia. By immunoblotting, these recognized several parathyroid proteins. No antibodies were observed against the CaSR or PTH. By RT PCR, AIRE mRNA was not found in the parathyroid. PMID- 14557430 TI - Efficacy and safety of oral weekly ibandronate in the treatment of postmenopausal osteoporosis. AB - Adherence to oral daily bisphosphonate regimens in postmenopausal osteoporosis is currently suboptimal. Less frequent dosing regimens are likely to improve patient adherence and thus, potentially, patient outcomes. A multicenter, randomized, double-blind, noninferiority study was conducted in 235 women (53-80 yr old; time since menopause >/==" BORDER="0"> 3 yr) with postmenopausal osteoporosis [lumbar spine (L1-L4) bone mineral density (BMD) T-score /==" BORDER="0"> 27 kg/m(2)) with PCOS were randomized to receive either metformin (500 mg twice daily for 3 months, then 1000 mg twice daily for 3 months) or ethinyl estradiol (35 micro g) cyproterone acetate (2 mg) oral contraceptive pills. The serum concentrations of CRP were significantly higher in obese than in nonobese subjects at baseline [4.08 +/- 0.53 (SE) vs. 1.31 +/- 0.28 mg/liter; P < 0.001] and correlated to BMI and to a lesser extent waist-hip ratio, suggesting that the elevated CRP levels may be related to obesity and not only to PCOS itself. During metformin treatment, serum CRP levels decreased significantly from 3.08 +/- 0.7 mg/liter to 1.52 +/- 0.26 mg/liter at 6 months in the whole study population (P = 0.006) and especially in obese subjects. In contrast, the treatment with ethinyl estradiol cyproterone acetate increased serum CRP levels from 2.91 +/- 0.68 mg/liter to 4.58 +/- 0.84 mg/liter (P < 0.001). Whether this effect is related to estrogen action in the liver or whether it reflects increased inflammation process and possible risks for cardiovascular disease remains unclear. The decrease of serum CRP levels during metformin therapy is in accordance with the known beneficial metabolic effects of this drug and suggests that CRP or other inflammation parameters could be used as markers of treatment efficiency in women with PCOS. PMID- 14557436 TI - Effects of an excess and a deficiency of endogenous parathyroid hormone on volumetric bone mineral density and bone geometry determined by peripheral quantitative computed tomography in female subjects. AB - Peripheral quantitative computed tomography (pQCT) is useful for evaluating volumetric bone mineral density (vBMD) as well as bone mineral density (BMD) of cortical and trabecular bones separately. Although PTH affects cortical and trabecular bones differently, the effects of endogenous PTH on vBMD and bone geometry have not previously been examined with pQCT. We, therefore, investigated the effects of an excess and a deficiency of endogenous PTH on bone by employing dual-energy x-ray absorptiometry and pQCT in 36 female patients with primary hyperparathyroidism (hyper), nine female patients with idiopathic or postoperative hypoparathyroidism (hypo), and 100 normal controls matched to age, gender, and body size (cont). Lumbar BMD by dual-energy x-ray absorptiometry was higher in the order: hypo > cont = hyper, and radius-1/3 BMD was significantly higher in the order: hypo > cont > hyper. The area of radius-1/3 was significantly higher in hyper than in cont. As for pQCT, trabecular vBMD was significantly higher in the order: hypo > cont > hyper at the 4% site (hypo, 157.5 +/- 36.7 mg/cm(3); cont, 123.4 +/- 47.5 mg/cm(3); hyper, 98.4 +/- 41.7 mg/cm(3)). Cortical vBMD was higher in the order: hypo > cont > hyper at the 20% site (hypo, 1141.1 +/- 53.1 mg/cm(3); cont, 1090.2 +/- 72.9 mg/cm(3); hyper, 1038.6 +/- 89.1 mg/cm(3)). Total bone area and endosteal and periosteal circumferences were significantly higher in hyper than in cont and hypo. Cortical area and thickness were higher in the order: hypo > cont > hyper. Bone strength indices were not significantly different among the three groups. In conclusion, vBMD evaluation revealed that an excess of endogenous PTH was catabolic for both cortical and trabecular bones, and that bone mass (especially trabecular bone mass) was preserved under a condition of deficient endogenous PTH. An excess of endogenous PTH stimulated periosteal bone formation, which might partly compensate for a decrease in bone strength induced by low BMD. PMID- 14557437 TI - Contraceptive efficacy of a depot progestin and androgen combination in men. AB - WHO studies provided proof of concept for hormonal male contraception using a prototype androgen-alone regimen. Combined testosterone plus progestin regimens offer more practical promise, but no contraceptive efficacy studies have been completed. The objective of this study was to establish the proof of principle for depot hormonal androgen/progestin combination as a male contraceptive. We performed a contraceptive efficacy study of 55 healthy men in stable fertile relationships seeking a change in contraceptive method. Testosterone (four 200-mg implants, every 4 or 6 months) and 300 mg depot medroxyprogesterone acetate, im, every 3 months were administered. Once sperm output was suppressed (<1 million/ml for 2 consecutive months), men entered a 12-month contraceptive efficacy period, ceasing other contraception. The main outcome measure was contraceptive failure (pregnancy) rate. No pregnancies occurred in 426 person-months (35.5 person years; 95% confidence limits for contraceptive failure rate, 0-8%/annum), superior to the first year failure rate of condoms, the only reversible male method. Sperm density fell rapidly, so 94% of men entered the efficacy phase by 3 months, with only 2 of 55 (3.6%) men not sufficiently suppressed to enter efficacy. A few men treated with testosterone implants at 6-month intervals demonstrated androgen deficiency symptoms and/or escape of gonadotropin and spermatogenic suppression between months 5 and 6; after a protocol amendment, all men receiving testosterone implants at 4-month intervals avoided androgen deficiency or loss of gonadotropin and sperm output suppression. Recovery was complete (median, 3.6 months to sperm reappearance and 5.0 months to 20 million sperm/ml) in all but one man with an incidental testicular disorder. Discontinuations were for protocol-related reasons (n = 15) or altered personal circumstances (n = 12), but there were no serious adverse effects related to drug exposure. The first male contraceptive efficacy study using a prototype depot androgen/progestin combination demonstrates high contraceptive efficacy with satisfactory short-term safety and recovery of spermatogenesis. Further studies of purpose-developed products are required to extend the overall safety and efficacy experience with depot androgen/progestin combinations, the most promising approach to hormonal male contraception. PMID- 14557438 TI - The impact of dose and route of estrogen administration on the somatotropic axis in normal women. AB - Oral estrogen therapy has reliably been found to reduce levels of serum IGF-I and increase mean 24-h GH levels in postmenopausal women as well as increase GH requirements in patients with GH deficiency. It is thought to act by inhibiting GH-stimulated IGF-I secretion, thus resulting in diminished feedback at the hypothalamic-pituitary axis and, hence, increased GH levels. In contrast, the administration of transdermal estrogen has variably been found to reduce, not change or increase, levels of serum IGF-I. We sought to clarify the effect of transdermal estrogen on the GH/IGF-I axis by using the IGF-I generation test, in which the acute response to a bolus dose of GH is examined. Nine healthy postmenopausal women received three different formulations of estrogen: oral estradiol (1 mg every 12 h), transdermal estradiol (50 micro g/d), and transdermal estradiol (200 micro g/d) for a 6-wk period in random order, separated by an 8-wk washout period. At the start of the study and in the last week of each estrogen formulation treatment, subjects underwent an IGF-I generation test. Oral estradiol reduced baseline (P < 0.05) and GH stimulated (P < 0.05) IGF-I levels, and GH stimulated IGF-binding protein-3 (IGFBP-3) levels (P < 0.05). High dose transdermal estrogen did not affect basal levels of IGF-I or IGFBP-3, but reduced the response of these GH-dependent peptides to GH stimulation (P < 0.05). Low dose transdermal estrogen did not alter either baseline or peak IGF-I levels, but reduced the peak IGFBP-3 response to GH stimulation (P < 0.05). Estradiol levels were lower during both transdermal estrogen preparations than during oral estrogen. It has been suggested that the effect of estrogen on responsiveness to GH is limited to that administered by the oral route. We have demonstrated that transdermal estrogen also has a significant impact on responsiveness to GH despite achieving levels of circulating estrogen lower than those achieved by oral estrogen replacement. PMID- 14557439 TI - Oxidative stress is associated with adiposity and insulin resistance in men. AB - To investigate the direct relationship of oxidative stress with obesity and insulin resistance in men, we measured the plasma levels of 8-epi-prostaglandin F2alpha (PGF2alpha) in 14 obese and 17 nonobese men and evaluated their relationship with body mass index; body fat weight; visceral, sc, and total fat areas, measured by computed tomography; and glucose infusion rate during a euglycemic hyperinsulinemic clamp study. Obese men had significantly higher plasma concentrations of 8-epi-PGF2alpha than nonobese men (P < 0.05). The plasma levels of 8-epi-PGF2alpha were significantly correlated with body mass index (r = 0.408; P < 0.05), body fat weight (r = 0.467; P < 0.05), visceral (r = 0.387; P < 0.05) and total fat area (r = 0.359; P < 0.05) in all (obese and nonobese) men. There was also a significant correlation between the plasma levels of 8-epi PGF2alpha and glucose infusion rate in obese men (r = -0.552; P < 0.05) and all men (r = -0.668; P < 0.01). In all subjects, the plasma levels of 8-epi-PGF2alpha were significantly correlated with fasting serum levels of insulin (r = 0.487; P < 0.01). In brief, these findings showed that the circulating levels of 8-epi PGF2alpha are related to adiposity and insulin resistance in men. Although correlation does not prove causation, the results of this study suggest that obesity is an important factor for enhanced oxidative stress and that this oxidative stress triggers the development of insulin resistance in men. PMID- 14557440 TI - Adult height and adult sitting height in childhood medulloblastoma survivors. AB - Outcome in adult height and sitting height is poor in children surviving medulloblastoma due to craniospinal irradiation (CSRT) and chemotherapy. We evaluated adult height and sitting height in 51 medulloblastoma patients stratified into four groups: G1, GH-deficient (GHD) patients treated with 23-39 Gy CSRT but not treated with GH [recombinant human (rh)GH]; G2, patients treated with rhGH; G3, patients who were not GHD; and G4, patients treated with 18 Gy CSRT and rhGH. Standing/sitting height of each group was compared with parental height and previously reported outcome studies. The rhGH dose was 0.3 mg/kg.wk, a higher dose compared with other reports of adult heights. The adult heights were significantly taller in group G2 [mean height SD score (SDS) = -1.86] than that achieved in previous studies (P < 0.0001), but not different from group G3, non GHD (mean SDS = -1.55). The tallest stature achieved was in group G4 (18 Gy CSRT), a height SDS of -1.01. Sitting heights were significantly less than the normal population, with mean SDS of -2.96 but -1.62 in group G4. We conclude that adult heights but not sitting heights in medulloblastoma survivors are significantly improved with the higher dose of rhGH. The lower dose of CSRT further improves not only adult height but also sitting height. PMID- 14557441 TI - Early endocrine, metabolic, and sonographic characteristics of polycystic ovary syndrome (PCOS): comparison between nonobese and obese adolescents. AB - Approximately half of all women with polycystic ovary syndrome (PCOS) are overweight or obese, and studies have reported endocrine and metabolic differences between lean and obese women with PCOS. PCOS has not been as extensively investigated in the adolescent population. The objectives of our study were to further characterize early endocrine and metabolic alterations in adolescents with PCOS and to determine whether differences between nonobese and obese women with PCOS are present early in its course. We studied an ethnically heterogeneous group of 48 adolescents: 11 nonobese with PCOS [age, 16.1 +/- 1.9 yr; body mass index (BMI), 22.5 +/- 1.5 kg/m(2)], 22 obese with PCOS (age, 15.5 +/- 1.4 yr; BMI, 35.9 +/- 6.2 kg/m(2)), and 15 obese controls (age, 14.4 +/- 1.5 yr; BMI, 35.8 +/- 7.1 kg/m(2)). Fasting levels of glucose, insulin, proinsulin, hemoglobin A1c, testosterone, SHBG, Delta4-androstenedione (Delta4-A), dehydroepiandrosterone sulfate (DHEAS), LH, FSH, IGF-I, IGF binding protein-1, free IGF-I, and lipids were measured. Six of the 11 nonobese PCOS subjects, 11 of the 22 obese PCOS subjects, and six of the 15 controls underwent standard oral glucose tolerance testing. The insulin response to the oral glucose tolerance test was measured by the insulin area under the curve (I(AUC120)). Measures of insulin sensitivity were calculated as the fasting glucose to insulin ratio, quantitative insulin sensitivity check index, and composite insulin sensitivity index. The nonobese adolescents with PCOS demonstrated higher levels of LH, SHBG, Delta4-A, DHEAS, dihydrotestosterone, free IGF-I, and high-density lipoprotein, and lower low-density lipoprotein, compared with the obese PCOS group. Fasting levels of insulin and proinsulin, I(AUC120), and log I(AUC120) were higher, and the fasting glucose to insulin ratio, quantitative insulin sensitivity check index, and composite insulin sensitivity index were lower in the obese compared with the nonobese PCOS subjects. Greater levels of LH and androgens, including total and free testosterone, Delta4-A, and DHEAS, and lower SHBG levels were found in the obese PCOS group compared with the obese controls. Adolescents with PCOS manifest clinical, metabolic, and endocrine features similar to those of adult women, and differences between nonobese and obese women with PCOS may be detected in adolescence. Our findings indicate a more pronounced alteration in the hypothalamo-pituitary-adrenal axis in nonobese adolescents with PCOS and a more marked dysregulation of insulin levels and impairment of insulin sensitivity in their obese counterparts. Our data also suggest differences in the IGF system between nonobese and obese adolescents with PCOS. PMID- 14557442 TI - Stability of sleep timing against the melatonin secretion rhythm with advancing age: clinical implications. AB - Aging is often associated with decreased ability of sleep maintenance. It has been hypothesized that the elderly experience a delayed timing of sleep period relative to the circadian phase of various sleep-promoting physiological functions, possibly causing decreased sleep propensity in the latter part of their nocturnal sleep. We evaluated the relationship between the sleep timing and circadian phase of melatonin secretion, which is known as a possible human sleep modulator as well as a stable marker of biological clock phase (BCP). Actigraph sleep recordings were performed, followed by the evaluation of melatonin phase under dim light in 42 healthy elderly volunteers (mean age, 68.8 yr; male/female ratio, 16/26) and 27 healthy young male volunteers (22.5 yr). Elderly subjects showed remarkable clock time advances in both the midpoint of BCP and sleep timing, with a significant decrease in sleep maintenance ability. However, they showed no significant age-related changes in the sleep timing against the midpoint of BCP, suggesting that early morning awakening in the elderly appeared in a BCP for which sleep propensity remained sufficient to sustain sleep. The present findings do not support the hitherto known hypothesis that age-related delay in the sleep timing against the BCP induces the deterioration in sleep maintenance in the elderly. PMID- 14557443 TI - Oxyntomodulin suppresses appetite and reduces food intake in humans. AB - Oxyntomodulin (OXM) is released from the gut postprandially, in proportion to energy intake, and circulating levels of OXM are elevated in several conditions associated with anorexia. Central injection of OXM reduces food intake and weight gain in rodents, suggesting that OXM signals food ingestion to hypothalamic appetite-regulating circuits. We investigated the effect of iv OXM (3.0 pmol/kg.min) on appetite and food intake in 13 healthy subjects (body mass index, 22.5 +/- 0.9 kg/m(2)) in a randomized, double-blind, placebo-controlled, cross over study. Infusion of OXM significantly reduced ad libitum energy intake at a buffet meal (mean decrease, 19.3 +/- 5.6%; P < 0.01) and caused a significant reduction in scores for hunger. In addition, cumulative 12-h energy intake was significantly reduced by infusion of OXM (mean decrease, 11.3 +/- 6.2%; P < 0.05). OXM did not cause nausea or affect food palatability. Preprandial levels of the appetite-stimulatory hormone, ghrelin, were significantly suppressed by OXM (mean reduction, 44 +/- 10% of postprandial decrease; P < 0.0001). Elevated levels of endogenous OXM associated with disorders of the gastrointestinal tract may contribute to anorexia. PMID- 14557444 TI - Evaluation of diagnostic accuracy of insulin-like growth factor (IGF)-I and IGF binding protein-3 in growth hormone-deficient children and adults using ROC plot analysis. AB - We critically evaluated the diagnostic value of IGF-I and IGF-binding protein-3 (IGFBP-3) in GH deficiency (GHD) in children and adults using receiver operating characteristic (ROC) plot analysis. Sixty-six children (chronological age, 1.3-15 yr) were studied: 34 GHD and 32 idiopathic short stature (ISS). Ninety-two adults (chronological age, 18-70 yr) were also evaluated: 72 GHD, 34 of childhood onset (AGHD-CO), and 38 of adult onset (AGHD-AO); and 20 healthy volunteers. The SD score (SDS) for IGF-I was calculated from 596 normal subjects (212 children and 384 adults), and the SDS for IGFBP-3 was calculated from 350 normal subjects (212 children and 138 adults). The ROC plot showed that the best IGF-I SDS cut-off line was -1.65 for children [sensitivity (S), 68%; specificity (Sp), 97%, diagnostic efficiency (DEf), 81%], the cut-off line for AGHD was -1.65 for AGHD CO (S, 91%; Sp, 100%; DEf, 94%), and the cut-off line for AGHD-AO was -1.80 (S, 81%; Sp, 100%; DEf, 88%). For IGFBP-3 SDS, the best cut-off line was -1.80 for children (S, 90%; Sp, 60%; DEf, 78%); it was -1.45 for AGHD-CO (S, 90%; Sp, 75%; DEf, 82%) and -0.90 for AGHD-AO (S, 90%; Sp, 68%; DEf, 77%). An accurate diagnosis was obtained using IGF-I SDS alone in GHD children 65%; ISS, 97%; AGHD CO, 92%; AGHD-AO, 86%, with IGFBP-3 SDS alone in GHD children 60%; ISS, 90%; AGHD CO, 75%; AGHD-AO, 68%. Considering both, an accurate diagnosis was obtained in GHD children 60%; ISS, 87%; AGHD-CO, 71%; AGHD-AO, 64%. In conclusion, our findings support the need to use cut-off lines expressed in SDS obtained using an appropriate statistical methodology for better characterization of the various clinical presentations. IGF-I proved to be more useful because of its good diagnostic efficiency and accuracy in both children and adults, whereas IGFBP-3 did not significantly contribute to the diagnosis of GHD. PMID- 14557446 TI - Flutamide-metformin therapy to reduce fat mass in hyperinsulinemic ovarian hyperandrogenism: effects in adolescents and in women on third-generation oral contraception. AB - Adolescents and young women with menstrual irregularities and hyperinsulinemic hyperandrogenism, so-called polycystic ovary syndrome, have abnormalities in body composition, even when nonobese. The combination of flutamide (125-250 mg/d) and metformin has additive benefits on endocrine- metabolic indices in women with polycystic ovary syndrome. However, it is unknown whether this combination also reverses the abnormalities in body composition in this population, especially if the flutamide dose is further reduced, the combination is given at a young age, and an oral contraceptive (OC) is added. Two randomized 3-month studies with flutamide-metformin were conducted in nonobese patients (n = 45) with hyperinsulinemic ovarian hyperandrogenism. In teenagers [n = 21; approximately 15 yr; no use of OC (OC-)], we determined whether a minidose combination (flutamide 62.5 mg/d) improved body composition and endocrine-metabolic indices, and in young women (n = 24; approximately 18 yr; OC+), we determined whether the addition of the minidose combination to a gestodene-containing OC exerted additive effects on the same variables. In OC- teenagers, flutamide-metformin improved fasting insulin/glucose ratio; serum IGF-binding protein-1, testosterone, SHBG, androstenedione, triglycerides, low- density lipoprotein and high-density lipoprotein cholesterol; lean mass, total fat, and abdominal fat (all P < 0.01). In OC+ women, the addition of flutamide-metformin was associated with gain of lean mass and loss of total fat (P < 0.01 vs. OC alone), but the addition failed to reduce abdominal fat. In conclusion, in nonobese adolescents with hyperinsulinemic ovarian hyperandrogenism, minidose flutamide-metformin reversed endocrine-metabolic anomalies and the excess of fat, including abdominal fat; in women, adding flutamide- metformin to a third-generation OC was also effective in improving body composition, but abdominal adiposity was not improved. PMID- 14557445 TI - Transsphenoidal surgery for pituitary tumors in the United States, 1996-2000: mortality, morbidity, and the effects of hospital and surgeon volume. AB - Larger surgical caseload is associated with better patient outcome for many complex procedures. We examined the volume-outcome relationship for transsphenoidal pituitary tumor surgery using the Nationwide Inpatient Sample, 1996-2000. Multivariate regression adjusted for patient demographics, acuity measures, medical comorbidities, and endocrine status. A total of 5497 operations were performed at 538 hospitals by 825 surgeons. Outcome measured at hospital discharge was: death (0.6%), discharge to long-term care (0.9%), to short-term rehabilitation (2.1%), or directly home (96.2%). Outcomes were better after surgery at higher-volume hospitals (OR 0.74 for 5-fold-larger caseload, P = 0.007) or by higher-volume surgeons (OR 0.62, P = 0.02). A total of 5.4% of patients were not discharged directly home from lowest-volume-quartile hospitals, compared with 2.6% at highest-volume-quartile hospitals. In-hospital mortality was lower with higher-volume hospitals (P = 0.03) and surgeons (P = 0.09). Mortality rates were 0.9% at lowest-caseload-quartile hospitals and 0.4% at highest-volume-quartile hospitals. Postoperative complications (26.5% of admissions) were less frequent with higher-volume hospitals (P = 0.03) or surgeons (P = 0.005). Length of stay was shorter with high-volume hospitals (P = 0.02) and surgeons (P < 0.001). Hospital charges were lower for high-volume hospitals, but not significantly. This analysis suggests that higher-volume hospitals and surgeons provide superior short-term outcomes after transsphenoidal pituitary tumor surgery with shorter lengths of stay and a trend toward lower charges. PMID- 14557447 TI - Clinical utility of an immunoradiometric assay for parathyroid hormone (1-84) in primary hyperparathyroidism. AB - The reliable diagnosis of primary hyperparathyroidism depends on the measurement of PTH. The PTH assays in widespread use measure not only the hormone but also hormone fragments, thus limiting the clinical utility of the assays. A new immunoradiometric assay (IRMA) using an antigenic determinant at the extreme amino-terminal of the PTH molecule detects only full-length PTH (1-84). We compared three PTH assays and determined the presence of PTH (1-84) and PTH fragments in serum and parathyroid adenomas of patients with primary hyperparathyroidism. We studied 56 patients with primary hyperparathyroidism. PTH levels were increased in 63% using the midmolecule RIA; in 73% in the "intact" IRMA; and in 96% in the PTH (1-84)-IRMA. The PTH (1-84)-IRMA correlated with the other assays (midmolecule RIA R = +0.736; P < 0.0001; "intact"-IRMA R = +0.951; P < 0.0001) and indices of disease activity (serum calcium R = +0.511, P < 0.0001; alkaline phosphatase R = +0.489, P = 0.001; and radius bone density R = -0.366, P < 0.01). In 21 consecutive patients undergoing parathyroidectomy, 18 had parathyroid adenomas. Intact PTH was higher than PTH (1-84)-IRMA in both serum and glandular homogenates from these patients. Similar proportions of PTH (1-84) and hormone fragments were found in both adenomas [66 +/- 3% of "intact" PTH reflected PTH (1-84) and sera (73 +/- 2% of "intact" PTH reflected PTH (1-84)]. We conclude that the PTH (1-84)-IRMA offers improved diagnostic sensitivity in patients with primary hyperparathyroidism than other currently available assays. This study also provides evidence that both PTH (1-84) and PTH fragments are produced in parathyroid adenomas and that peripheral metabolism of hormone and fragment does not alter the proportion of bioactive hormone. PMID- 14557448 TI - Final height and body mass index among adult survivors of childhood brain cancer: childhood cancer survivor study. AB - The objectives of this study were 1) to compare final height and body mass index (BMI) between adult survivors of childhood brain cancer and age- and sex-matched population norms, 2) to quantify the effects of treatment- and cancer-related factors on the risk of final height below the 10th percentile (adult short stature) or having a BMI of 30 kg/m(2) or more (obesity). Treatment records were abstracted and surveys completed by 921 adults aged 20-45 yr who were treated for brain cancer as children and were participants in the multicenter Childhood Cancer Survivor Study. Nearly 40% of childhood brain cancer survivors were below the 10th percentile for height. The strongest risk factors for adult short stature were young age at diagnosis and radiation treatment involving the hypothalamic-pituitary axis (HPA). The multivariate odds ratio for adult short stature among those 4 yr of age or younger at diagnosis, relative to ages 10-20 yr, was 5.67 (95% confidence interval, 3.6-8.9). HPA radiation exposure increased the risk of adult short stature in a dose-response fashion (trend test, P < 0.0001). Adjuvant chemotherapy was not an independent risk factor for adult short stature. BMI distribution in survivors did not differ appreciably from that of population norms; however, in females, young age at diagnosis and HPA radiation dose (trend test, P < 0.001) were associated with risk of obesity. Except for patients treated with surgery only, survivors of childhood brain cancer are at very high risk for adult short stature, and this risk increases with radiation dose involving the HPA. We did not find a corresponding elevated risk for obesity. PMID- 14557449 TI - Soy isoflavones have a favorable effect on bone loss in Chinese postmenopausal women with lower bone mass: a double-blind, randomized, controlled trial. AB - Animal studies have shown that soy isoflavones have an effect in preventing estrogen-related bone loss, but few data are available in humans, especially in the Asian populations. This double-blind, placebo-controlled, randomized trial examines the effects of soy isoflavones on bone loss in postmenopausal Chinese women, aged 48-62 yr. Two hundred and three eligible subjects were randomly assigned to three treatment groups with daily doses of placebo (1 g starch; n = 67), mid-dose (0.5 g starch, 0.5 g soy extracts, and approximately 40 mg isoflavones; n = 68), and high dose (1.0 g soy extracts and approximately 80 mg isoflavones; n = 68). All were given 12.5 mmol (500 mg) calcium and 125 IU vitamin D(3). Bone mineral density (BMD) and bone mineral content (BMC) of the whole body, spine, and hip were measured using dual energy x-ray absorptiometry at baseline and 1 yr post treatment. Both univariate and multivariate analyses showed that women in the high dose group had mild, but statistically significantly, higher favorable change rate in BMC at the total hip and trochanter (P < 0.05) compared with the placebo and mid-dose groups, even after further adjustments for the potential confounding factors. Further stratified analyses revealed that the positive effects of soy isoflavone supplementation were observed only among women with lower initial baseline BMC (median or less). In conclusion, soy isoflavones have a mild, but significant, independent effect on the maintenance of hip BMC in postmenopausal women with low initial bone mass. PMID- 14557450 TI - The mathematical model for total pubertal growth in idiopathic growth hormone (GH) deficiency suggests a moderate role of GH dose. AB - The role of GH treatment during total pubertal growth (TPG) is still unclear. We developed a prediction model for TPG (centimeters) through a multiple regression analysis of various prepubertal parameters in 303 adolescents with idiopathic GH deficiency from the KIGS database. Prepubertal catch-up growth and near-adult height were achieved, and GH dose was kept constant at approximately 30 micro g/kg.d. The model was validated on a cohort of 36 patients from one center. Four TPG predictors explained 70% of the variability with an error SD of 4.2 cm: gender (TPG in males was >11.3 cm vs. that in females), age at onset of puberty (negative), height SD score minus midparental height SD score at puberty onset (negative), and mean GH dose during puberty (positive). Our analysis suggests that TPG in idiopathic GH deficiency is only moderately dependent on GH dose. The use of a higher GH dosage at the onset of puberty should thus depend on the individual's height development. The TPG model aids in the planning of individually optimized and cost-effective GH treatment. PMID- 14557451 TI - A reappraisal of the utility of somatostatin receptor scintigraphy in patients with ectopic adrenocorticotropin Cushing's syndrome. AB - Ectopic ACTH hypersecretion is a rare cause of Cushing's syndrome. Bronchial carcinoids are the most common neoplasms causing the occult ectopic ACTH syndrome (EAS). Localization of these tumors is often difficult. The diagnostic utility of somatostatin receptor scintigraphy (SRS) in EAS has been studied in a limited number of patients with conflicting results. Herein we report our experience with 12 consecutive cases. Histological confirmation was obtained in nine patients, the majority being bronchial carcinoids. Among the seven patients with histologically confirmed bronchial carcinoids, SRS was performed in six patients. In three patients SRS correctly localized a bronchial carcinoid tumor at presentation. In the remaining three it became positive after 8, 22, and 27 months during follow-up. In two patients SRS was positive without any finding in the corresponding conventional imaging study. In two patients positive computed tomography/magnetic resonance imaging preceded SRS localization. There was no false positive SRS. Among three patients with highly suspected EAS, SRS was positive in one. Both patients with EAS due to medullary thyroid carcinoma had focal positive uptake. In summary, in this study a substantial number of patients had positive tumor localization by SRS. Therefore, SRS is a useful tool in the evaluation of patients with EAS. PMID- 14557452 TI - Optimizing control of acromegaly: integrating a growth hormone receptor antagonist into the treatment algorithm. AB - Acromegaly is associated with significant morbidities and a 2- to 3-fold increase in mortality because of the excessive metabolic action of GH and IGF-I, a marker of GH output. Reductions in morbidity correspond with decreases in IGF-I, and mortality is lowered following normalization of IGF-I or GH levels. Therefore, this has become an important end point. Current guidelines for the treatment of acromegaly have not considered recent advances in medical therapy, in particular, the place of pegvisomant, a GH receptor antagonist. Treatment goals include normalizing biochemical markers, controlling tumor mass, preserving pituitary function, and relieving signs and symptoms. Surgery reduces tumor volume and is considered first-line therapy. Radiation reduces tumor volume and GH and IGF-I levels, but the onset of action is slow and hypopituitarism typically develops. Therefore, pharmacotherapy is often used following surgery or as first-line therapy for nonresectable tumors. Dopamine agonists can be considered in patients exhibiting minimal disease or those with GH-prolactin-cosecreting tumors but will not achieve hormone normalization in most patients. Somatostatin analogs effectively suppress GH and IGF-I in most patients, but intolerance (e.g. diarrhea, cramping, gallstones) can occur. Pegvisomant, the newest therapeutic option, blocks GH action at peripheral receptors, normalizes IGF-I levels, reduces signs and symptoms, and corrects metabolic defects. Pegvisomant does not appear to affect tumor size and has few adverse effects. Pegvisomant is the most effective drug treatment for acromegaly in normalizing IGF-I and producing a clinical response; it is the preferred agent in patients resistant to or intolerant of somatostatin analogs. PMID- 14557453 TI - Unique epitopes of glutamic acid decarboxylase autoantibodies in slowly progressive type 1 diabetes. AB - Disease-specific epitope profiles of glutamic acid decarboxylase (GAD)65 autoantibodies (GAD65Ab) were studied in slowly progressive type 1 (insulin dependent) diabetes mellitus (SPIDDM) and acute onset type 1 (insulin-dependent) diabetes mellitus (AIDDM) using seven kinds of GAD65/67 chimeric molecules. Sera obtained from Japanese SPIDDM (n = 17) and AIDDM (n = 46) patients followed prospectively were analyzed by immunoprecipitation, ELISA, and Western blotting. GAD65Ab in all SPIDDM samples reacted specifically with an N-terminal linear epitope located on the membrane anchoring domain between amino acids 17-51 and C terminal conformational epitope between amino acids 443-585 of GAD65. The binding of GAD65Ab with N-terminal 83 residues in SPIDDM inversely correlated with the period in which insulin was not required. GAD65Ab in AIDDM did not react with N terminal epitope located between amino acids 1-83, irrespective of the titer of GAD65Ab. A novel epitope of GAD65Ab in AIDDM residing between amino acids 244-360 was identified in 17% (8 of 46) of patients whose age of onset was younger than other AIDDM patients. In conclusion, GADAb in SPIDDM has unique N-terminal linear epitopes that are located on the anchoring domain of GAD65 molecules. Association is suggested between GAD65Ab targeted to this region and slowly progressive beta cell failure in SPIDDM. PMID- 14557455 TI - Regulatory effects of gonadotropin-releasing hormone (GnRH) I and GnRH II on the levels of matrix metalloproteinase (MMP)-2, MMP-9, and tissue inhibitor of metalloproteinases-1 in primary cultures of human extravillous cytotrophoblasts. AB - An intricate balance between the production of matrix metalloproteinases (MMPs) and their endogenous inhibitors, tissue inhibitors of metalloproteinases (TIMPs), modulates the overall proteolytic activity of trophoblasts during human implantation. In these studies we have examined the ability of classical GnRH I and the second form of this hormone (GnRH II) to regulate MMP-2, MMP-9, and TIMP 1 mRNA and protein levels in extravillous cytotrophoblasts propagated from explants of first trimester chorionic villi. GnRH I and GnRH II were found to increase MMP-2 and MMP-9 mRNA and protein levels in these primary cell cultures in a dose- and time-dependent manner using quantitative competitive-PCR and ELISA. In contrast, these two hormones decreased trophoblastic TIMP-1 mRNA and protein levels. Cetrorelix, a GnRH receptor antagonist, inhibited the regulatory effects of GnRH I, but not GnRH II, on MMP-2, MMP-9, and TIMP-1 expression in these cells. Collectively, these observations suggest that GnRH I and GnRH II differentially regulate MMP-2, MMP-9, and TIMP-1 expression in human trophoblasts, possibly via distinct receptor-mediated intracellular signaling pathways. PMID- 14557454 TI - Sexual dimorphism of growth hormone (GH) regulation in humans: endogenous GH releasing hormone maintains basal GH in women but not in men. AB - GH secretory patterns in humans are sexually dimorphic in terms of pulse regularity, amplitude of the diurnal rhythm, and magnitude of basal (trough) secretion. The neuroendocrine mechanisms of gender-specific GH regulation in humans are currently unknown, but the interpulse GH levels are generally assumed to be controlled by somatostatin. In rats, however, administration of antiserum to GHRH lowers GH interpulse levels in females but not males. In this study, using a competitive antagonist to GHRH in humans, we investigated whether endogenous GHRH has differential, gender-specific effects on the interpulse GH levels. Six healthy men and five healthy women (20-28 yr old) who were nonobese, did not smoke, and were on no medications known to influence GH secretion were studied. Each served as his or her own control during an infusion of GHRH antagonist or saline for a 27-h period. A control bolus of GHRH was given near the end of the infusion. In both sexes during GHRH antagonist infusion, mean GH, pulse amplitude, and GH response to GHRH decreased significantly, whereas pulse frequency remained unchanged. However, during the GHRH antagonist infusion, trough GH did not significantly change in men (P = 0.54) but significantly decreased in women (P = 0.008). Deconvolution analysis confirmed the lack of a significant change in basal secretion in men (P = 0.81) as opposed to women (P = 0.006). We conclude that sexual dimorphism in the neuroendocrine regulation of GH secretion in humans involves a differential role of endogenous GHRH in maintaining baseline GH. PMID- 14557456 TI - Insulin sensitivity, insulin secretion, and glucose tolerance versus intima-media thickness in nondiabetic postmenopausal women. AB - To study the association between insulin sensitivity and secretion vs. early manifestations of atherosclerosis, we performed a 5-yr prospective study in 84 nondiabetic, postmenopausal women, aged 58.7 +/- 0.4 yr (mean +/- SD). Insulin sensitivity was measured with the euglycemic, hyperinsulinemic clamp, and insulin secretion was measured as the acute response to iv arginine (5 g). Early atherosclerosis was studied by ultrasonography of the right carotid artery. Mean intima-media thickness (IMT), determined 1 cm proximal to the bifurcation, was 0.81 +/- 0.14 mm at baseline and increased by 0.012 +/- 0.014 mm/yr over the 5 yr (P < 0.001). The maximal IMT, determined in the carotid bifurcation, was 1.42 +/- 0.42 mm at baseline and increased by 0.035 +/- 0.049 mm/yr (P < 0.001). Neither basal IMT nor the increase in mean or maximal IMT correlated to insulin sensitivity or secretion. In contrast, both baseline IMT and the progression in IMT over the 5-yr follow-up (both mean common carotid artery IMT and maximal bifurcation IMT) correlated with systolic blood pressure and low-density lipoprotein cholesterol. We conclude that carotid intima-media thickness is not related to insulin sensitivity or secretion in nondiabetic, postmenopausal women. Instead, the strongest association is seen with systolic blood pressure and low density lipoprotein cholesterol levels. PMID- 14557457 TI - Effect of ingested nutrients on the release of thrittene into the human circulation. AB - Thrittene is a recently described peptide with a sequence homologous with somatostatin-28 ((1-13)) but is produced independent of the preprosomatostatin gene. It is localized in epithelial cells in stomach and gut mucosal crypts and in neuronal cell bodies in the myenteric plexus and enteric axons. It is also present in human plasma. The aim of this study was to determine whether the release of thrittene into the circulation was affected by the ingestion of nutrients and, if so, whether the pattern of release was distinct from the closely related peptide somatostatin-28 (S-28). Thrittene was indirectly measured in human plasma by an RIA using antiserum F4. F4 interacts with the Asn(5)-Pro(6) region shared by S-28 and thrittene. The contribution of S-28 to F4 immunoreactivity (F4-IR) was determined using a specific two-site assay, and this measure was subtracted from the total F4-IR to give an estimate of thrittene levels. Plasma for assay was taken from healthy men on 4 separate days before and after intake of: a mixed meal (715 kcal), and meals containing primarily fat (25 g; 225 kcal), carbohydrate (100 g; 454 kcal), and protein (22 g; 100 kcal). After the mixed meal, both S-28 and thrittene rose by 50-100% within 30 min and gradually declined by 4 h. These increments were mimicked after ingestion of fat. By contrast, thrittene levels increased after carbohydrate but not protein intake, whereas S-28 concentrations rose after protein but not carbohydrate ingestion. These findings indicate that thrittene is secreted into the mammalian circulation during food intake and raise the possibility that thrittene may play a role in nutrient disposition. The dichotomy in responses of thrittene and S-28 to ingestion of the major macronutrients suggests that they are secreted from different gastrointestinal cells. PMID- 14557458 TI - Association between insulin-like growth factor I (IGF-I) polymorphisms, circulating IGF-I, and pre- and postnatal growth in two European small for gestational age populations. AB - The purpose of this study was to assess the association of IGF-I and birth size by studying small for gestational age (SGA) subphenotypes and undertaking more detailed analysis of IGF-I genetic markers. SGA subjects from Haguenau, France (n = 113), and Gothenburg, Sweden (n = 174), were studied. The Swedish subjects were subphenotyped according to postnatal growth (114 short SGA and 60 SGA catch-up). IGF-I dinucleotide repeat and single nucleotide polymorphism (SNP) markers were studied, and haplotypes were generated in the Swedish short SGA group by identity of state. Association analysis was undertaken using the Monte Carlo method of association analysis of multiallelic markers for dinucleotide repeat markers, by exact chi(2) analysis for SNPs and by ANOVA for serum IGF-I levels. IGF-I genotype was associated with the SGA phenotype, in particular with symmetrical SGA and low birth weight, and with IGF-I levels in SGA subjects. Association with postnatal growth was different in the two populations, which may reflect the power of the smaller subphenotype groups. Haplotype analysis in the Swedish short SGA subjects showed that the region of association lay between the promoter and intron 2 of the IGF-I gene. These studies validate the association of the IGF-I gene with birth size and refine the region of association in Swedish short SGA subjects. PMID- 14557459 TI - Possible angiogenic roles of insulin-like growth factor II and its receptors in uterine vascular adaptation to pregnancy. AB - Adaptation of the maternal uterine vasculature is essential for normal fetal and placental development in which angiogenesis is considered one of the most critical adaptive changes during pregnancy. Highly expressed in cytotrophoblasts and maternal endothelial cells during pregnancy, IGF-II promotes cell migration and regulates fetal and placental growth. We hypothesized that IGF-II regulates uterine angiogenesis during pregnancy. Both uterine vasculature and isolated uterine microvascular endothelial cells expressed high levels of IGF-II and IGF II/mannose-6 phosphate receptor mRNA as shown by in situ hybridization. Physiological concentrations of IGF-II significantly increased vessel formation, as shown by a three-dimensional angiogenesis assay in vitro or a chicken chorionallantoic membrane assay in vivo. The angiogenic response of IGF-II could be reversed by the addition of beta-galactosidase or rabbit-antihuman IGF-II/M6P receptor antiserum, whereas blocking antibodies against IGF-I receptor or insulin receptor influenced IGF-II-induced sprout formation. IGF-II promoted migration of endothelial cells (10-250 ng/ml) tested in a modified Boyden chamber, but no stimulating effect on proliferation was observed. The application of several intracellular signal transduction molecules and their inhibitors indicated that protein kinase C and G(i) protein might play a role in the IGF-II-induced angiogenesis. Our results suggest an important angiogenic role of IGF-II in the vascular adaptation to pregnancy. PMID- 14557460 TI - Recombinant human thyrotropin reduces serum vascular endothelial growth factor levels in patients monitored for thyroid carcinoma even in the absence of thyroid tissue. AB - In this study, we have investigated in vivo the time-dependent effects of TSH on vascular endothelial growth factor (VEGF) production in patients monitored for thyroid carcinoma. Serum VEGF, thyroglobulin (Tg), and TSH levels were assayed at baseline and 6, 24, 30, 48, 72, and 96 h and 1 wk after administration of recombinant human TSH (rhTSH) in 45 thyroidectomized patients affected by differentiated thyroid carcinoma. At baseline, the patients with metastasis (18 cases) showed serum Tg and VEGF values significantly higher than those seen in the cured patients (27 cases). During rhTSH stimulation, the mean VEGF levels decreased significantly in both patient groups. In 60% of patients with metastasis, VEGF nadir occurred at the same time as serum TSH reached the highest values, whereas in 85.7% of the cured patients VEGF decreased after the TSH peak (P = 0.003). In conclusion, we demonstrate for the first time that short-term administration of rhTSH in patients monitored for differentiated thyroid carcinoma induces a significant reduction in serum VEGF values even in the absence of thyroid tissue. This result would suggest that TSH may be able in vivo to regulate VEGF production from tissues other than the thyroid gland. PMID- 14557461 TI - Serum adiponectin levels are inversely associated with overall and central fat distribution but are not directly regulated by acute fasting or leptin administration in humans: cross-sectional and interventional studies. AB - Adiponectin is an adipocyte-secreted protein that circulates in high concentrations in the serum and acts to increase insulin sensitivity. Previous studies have shown that serum adiponectin is inversely associated with fat mass and insulin resistance in humans and that acute fasting decreases adipose tissue adiponectin mRNA expression in rodents. Whether acute energy deprivation, body fat distribution, or serum hormone levels are associated with circulating adiponectin in humans remains largely unknown. To identify predictors of serum adiponectin levels, we evaluated the association of adiponectin with several anthropometric, metabolic, and hormonal variables in a cross-sectional study of 121 women without a known history of diabetes. We also performed interventional studies to assess whether fasting for 48 h and/or leptin administration regulates serum adiponectin in healthy men and women. Our cross-sectional study shows that, in addition to overall obesity, central fat distribution is an independent negative predictor of serum adiponectin and suggests that adiponectin may represent a link between central obesity and insulin resistance. In addition, estradiol is negatively and independently associated with adiponectin, whereas there is no association between serum adiponectin and leptin, cortisol, or free testosterone levels. Our interventional studies demonstrate that neither fasting for 48 h, resulting in a low leptin state, nor leptin administration at physiological or pharmacological doses alters serum adiponectin levels. Further studies are needed to fully elucidate the physiology of adiponectin in humans and its role in the pathogenesis of insulin-resistant states. PMID- 14557462 TI - Enhanced repression by HESX1 as a cause of hypopituitarism and septooptic dysplasia. AB - HESX1 is a paired-like homeodomain transcription factor that functions as a repressor of PROP1-mediated gene stimulation. Mutations in HESX1 have been implicated in cases of septooptic dysplasia and congenital hypopituitarism. All mutations in HESX1 identified to date have resulted in impaired DNA binding and defective HESX1 action. We have identified a novel HESX1 mutation in genomic nucleotide position 1684 (g.1684delG), which results in a mutant protein with increased DNA binding. In turn, this mutation causes increased repression of PROP1-dependent gene activity. These data suggest that enhancement of transcriptional repression during pituitary organogenesis is a novel mechanism for the development of congenital pituitary disorders. PMID- 14557463 TI - Phenotypic and genetic heterogeneity in congenital generalized lipodystrophy. AB - Congenital generalized lipodystrophy (CGL) is a rare autosomal recessive disorder characterized by near complete absence of adipose tissue from birth. Recently, mutations in 1-acylglycerol-3-phosphate O-acyltransferase 2 (AGPAT2) and Berardinelli-Seip congenital lipodystrophy 2 (BSCL2) genes were reported in pedigrees linked to chromosomes 9q34 and 11q13, respectively. There are limited data regarding phenotypic differences between the various subtypes of CGL. Furthermore, whether there are additional loci for CGL remains unknown. Therefore, we genotyped 45 pedigrees with CGL for AGPAT2 and BSCL2 loci and compared the phenotypes in the various subtypes. Twenty-six pedigrees harbored mutations, including seven novel variants, in the AGPAT2 gene, and 11 pedigrees harbored mutations in the BSCL2 gene, including five novel variants. Eight pedigrees had no substantial alterations in either gene. Of these, three informative pedigrees showed no linkage to markers spanning the AGPAT2 and BSCL2 loci, and in six of the affected subjects, the transcripts of AGPAT2 and BSCL2 were normal. All subtypes of CGL showed high prevalence of diabetes, hypertriglyceridemia, and acanthosis nigricans. However, patients with BSCL2 mutations had lower serum leptin levels, an earlier onset of diabetes, and higher prevalence of mild mental retardation compared with other subtypes. We conclude that besides AGPAT2 and BSCL2, there may be additional loci for CGL. The genetic heterogeneity in CGL patients is accompanied by phenotypic heterogeneity. PMID- 14557464 TI - Circulating resistin levels are not associated with obesity or insulin resistance in humans and are not regulated by fasting or leptin administration: cross sectional and interventional studies in normal, insulin-resistant, and diabetic subjects. AB - Resistin is a novel adipocyte-secreted hormone proposed to link obesity with diabetes. Studies in mice have revealed conflicting data however, and the physiological role of circulating resistin in humans remains unknown. We conducted cross-sectional studies in 123 middle-aged women and 120 healthy young subjects and found that serum resistin levels did not correlate with markers of adiposity, including body mass index, waist-to-hip ratio, or fat mass, or insulin resistance assessed by homeostasis model, lipid profile, or serum leptin levels; but females had higher resistin levels than males (P < 0.02). We also found no difference in serum resistin levels between lean healthy and obese insulin resistant nondiabetic and type 2 diabetic adolescents. Finally, to evaluate the effect of food deprivation and/or leptin administration on resistin levels, we performed interventional studies that revealed no significant difference in resistin levels after 48 h of fasting and/or leptin administration at either physiological or pharmacological doses. We conclude that circulating resistin is unlikely to play a major role in insulin resistance or energy homeostasis in humans. PMID- 14557465 TI - Elevated levels of mannan-binding lectin in patients with type 1 diabetes. AB - The hepatic protein mannan-binding lectin (MBL) activates the complement system on binding to carbohydrate patterns and is involved in first-line defense against invading microorganisms. Emerging evidence indicates that in some situations MBL may cause inexpedient complement activation and tissue injury through binding to endothelial glycosylations. MBL levels are suppressed by insulin treatment in critically ill patients, and, hypothetically, hepatic portal hypoinsulinemia could lead to increased levels of MBL in patients with type 1 diabetes. We measured MBL and C-reactive protein (CRP) levels in 132 normoalbuminuric type 1 diabetic patients and 66 healthy age- and sex-matched controls. The median MBL concentration was higher in diabetic patients than in healthy controls [1290 micro g/liter (interquartile range, IQR 354-2961 micro g/liter) vs. 970 micro g/liter (IQR 277-1607 micro g/liter), P = 0.025], whereas CRP concentrations were similar among patients and controls [1.42 mg/liter (IQR 0.95-2.21) vs. 1.21 mg/l (IQR 0.74-2.13), NS]. In diabetic subjects, CRP levels correlated with poor glycemic control as indicated by hemoglobin A(1c) and daily insulin dose, which was not the case with MBL. MBL concentrations were positively correlated with urinary albumin excretion (r = 0.22; P = 0.013) and increased with increasing urinary albumin excretion tertile (P = 0.036). In conclusion, our data demonstrate that circulating MBL concentrations are significantly elevated in patients with type 1 diabetes and suggest a possible role of MBL in the pathogenesis of renovascular complications in diabetes. PMID- 14557466 TI - The Gly16-->Arg16 and Gln27-->Glu27 polymorphisms of beta2-adrenergic receptor are associated with metabolic syndrome in men. AB - Endogenous catecholamines contribute to regulation of adipose tissue lipolysis, glucose homeostasis, and vascular tone. The goal of the present study was to assess the association between the Gly(16)-->Arg(16) and Gln(27)-->Glu(27) polymorphisms of the beta(2)-adrenergic receptor and metabolic syndrome. Participants were recruited in a population survey and included 1195 men and women. Metabolic syndrome was defined according to National Cholesterol Education Program Adult Treatment Panel III guidelines. There were 276 patients with metabolic syndrome and 872 controls. The Gly(16)-->Arg(16) (P < 0.005) and Gln(27)-->Glu(27) (P < 0.04) polymorphisms were associated with metabolic syndrome in men, but not in women. In multivariate analyses adjusting for age, physical activity, smoking habits, alcohol consumption, and body mass index, the odds ratio of metabolic syndrome was 1.83 (95% confidence interval, 1.10-3.05) and 2.43 (95% confidence interval, 1.19-4.95) in men bearing the Gly(16)/Arg(16) and Arg(16)/Arg(16) genotypes, respectively. Similarly, the odds ratios of metabolic syndrome were 0.99 (95% confidence interval, 0.50-1.93) and 1.67 (95% confidence interval, 0.84-3.33) in men bearing the Gln(27)/Glu(27) and Gln(27)/Gln(27) genotypes, respectively. Because both variants were in linkage disequilibrium, a haplotype analysis was performed. There was no evidence of any statistically significant association between beta(2)-adrenergic receptor haplotypes and metabolic syndrome. In conclusion, these data suggest that the Arg(16) and Gln(27) variants of the beta(2)-adrenergic receptor gene contribute to metabolic syndrome susceptibility in men. PMID- 14557467 TI - Octreotide abolishes the acute decrease in bone turnover in response to oral glucose. AB - Feeding or oral intake of glucose results in an acute suppression of bone turnover. This does not appear to be mediated by insulin. Several gastrointestinal hormones modulate bone turnover in vitro and may mediate this response. We examined whether inhibiting the production of gastrointestinal hormones using octreotide could block glucose-mediated suppression of bone turnover. Fifteen subjects were each studied on four occasions in a randomized, single-blind, crossover study after receiving 1) oral placebo, iv saline; 2) oral glucose, iv saline; 3) oral glucose, iv octreotide; or 4) iv octreotide alone. We measured serum C-terminal telopeptide of type I collagen, urinary N-terminal telopeptide of type I collagen, osteocalcin, procollagen type I N-terminal propeptide, PTH, insulin, ionized calcium, and glucose over 4 h. All bone turnover markers decreased significantly after oral glucose (P < 0.001). At 120 min serum C-terminal telopeptide decreased by 45 +/- 2%, urinary N-terminal telopeptide by 31 +/- 7%, osteocalcin by 16 +/- 1%, and procollagen type I N terminal propeptide by 8 +/- 1%. There was no significant decrease in bone turnover in response to oral glucose during octreotide infusion. Octreotide alone resulted in a significant increase in all bone turnover markers (P < 0.05) and PTH (P < 0.01). We conclude that octreotide completely abolishes the bone turnover response to glucose intake and increases PTH secretion. The apparent bone turnover response to feeding is probably mediated by an octreotide inhibitable endocrine factor. PMID- 14557468 TI - Leptin administration increases nocturnal concentrations of luteinizing hormone and growth hormone in juvenile female rhesus monkeys. AB - The importance of leptin in regulating sexual maturation is supported by data showing that deletions of the leptin gene or alterations in the leptin receptor result in infertility. However, attempts to define a role for leptin in normal puberty have produced equivocal results, leading to the conclusion that, if leptin is involved in puberty, its role is permissive and not obligatory. To better define the importance of leptin in primate puberty, the present study tested the hypothesis that a premature elevation in nocturnal leptin concentrations would accelerate indices of puberty, including nocturnal LH secretion in female rhesus monkeys (Macaca mulatta). Juvenile, gonadally intact females were treated daily with leptin (n = 6; 30 micro g/kg, sc at 1700 h) from 12-30 months of age and were compared with age-matched control females (n = 13). Chronic elevation in peripheral concentrations of leptin increased serum levels of both daytime and nighttime bioactive LH at a significantly younger age compared with control females. The earlier rise in LH in leptin-treated females was associated with an earlier increase in serum estradiol and occurrence of menarche. Despite this effect of leptin, nocturnal serum LH was significantly higher at each age assessed in non-leptin-treated ovariectomized controls (n = 6). In addition, leptin increased skeletal lengths and maturity that were associated with significantly higher serum levels of nocturnal GH and daytime IGF I. Although body weights were not consistently affected by treatment, body mass index, as an index of body fat, was consistently lower in leptin-treated females. Taken together, these data indicate that the chronic elevation in serum leptin concentrations advances the nocturnal increase in serum LH as well as other parameters of female puberty. Furthermore, the observation that nocturnal LH was higher in age-matched, agonadal females compared with the leptin-treated females suggests that the nongonadal drive to LH secretion is operative in female macaques as early as 14 months of age, suggesting that the effect of leptin on puberty in female primates may involve a diminution in gonadal negative feedback suppression of LH secretion. Such a role would suggest that leptin is permissive yet critical for advancing female puberty. PMID- 14557469 TI - Adiposity contributes to differences in left ventricular structure and diastolic function with age in healthy men. AB - We sought to examine the influence of adiposity in age-associated changes in the left ventricle (LV) in a cohort of 113 healthy men, aged 20-79 yr, by measuring LV structure and diastolic function (echocardiography), whole body composition, and regional adiposity (dual energy x-ray absorptiometry). Aging was associated with increased levels of adiposity, greater wall thickness to chamber radius ratio, LV concentric remodeling, and reduced LV diastolic function (all P < 0.05). Bivariate correlation analysis showed that mean LV wall thickness, a concentric LV morphology, and diastolic function were related to adiposity (r = 0.63 to 0.51; all P < 0.05). The relation between age and both mean LV wall thickness and concentric remodeling was reduced after controlling for percentage total body fat (by 38% and 54%, respectively), percentage abdominal fat (by 42% and 62%), and the abdominal/thigh fat ratio (by 35% and 46%). The diastolic function-age relation was reduced after controlling for percentage total body fat (by 35%), percentage abdominal fat (by 39%), and the abdominal/thigh fat ratio (by 29%). There were no apparent differences in the contribution of percentage total body fat, percentage abdominal fat, or abdominal/thigh fat to the association between age and LV structure/diastolic function. We conclude that increasing adiposity contributes to the LV remodeling/reduced diastolic function that occurs with aging in healthy men. PMID- 14557470 TI - Auxology is a valuable instrument for the clinical diagnosis of SHOX haploinsufficiency in school-age children with unexplained short stature. AB - SHOX (short stature homeobox-containing gene) mutations causing haploinsufficiency have been reported in some individuals with idiopathic short stature and in many patients with Leri-Weill-dyschondrosteosis. Around 80% of SHOX mutations are complete gene deletions, whereas diverse point mutations account for the rest. The aim of this study was to estimate the prevalence of SHOX mutations in children with idiopathic short stature and to give an unbiased characterization of the haploinsufficiency phenotype of such children. We recruited 140 children (61 girls), in our clinic, with idiopathic short stature, which was defined by the presence of normal IGF-I and free T(4); a normal karyotype in females; the absence of endomysium antibodies, of chronic organic, psychological, or syndromatic disease; and by the lack of clear signs of any osteodysplasia. Height, arm span, and sitting height were recorded, and subischial leg length was calculated. Two highly polymorphic microsatellite markers located around the SHOX coding region (CA-SHOX repeat and DXYS233) were PCR-amplified with fluorescent primers and separated in an automatic sequencing machine. Analysis of parental DNA was performed in the probands who had only one fragment size of each of both markers. SHOX haploinsufficiency caused by a SHOX deletion was confirmed in three probands (2%), all females, who carried a de novo deletion through loss of the paternal allele. Their auxological data revealed a significant shortening of arms and legs in the presence of a low-normal sitting height, when compared with the other 137 children tested. Therefore, the extremities-trunk ratio (sum of leg length and arm span, divided by sitting height) for total height was significantly lower in the three SHOX haploinsufficient probands, in comparison with the whole group. This observation was confirmed with the auxological data of five additional patients (four females) previously diagnosed with SHOX haploinsufficiency; all but the youngest girl had height-adjusted extremities-trunk ratios more than 1 SD below the mean. All children with SHOX haploinsufficiency exhibited at least one characteristic radiological sign of Leri-Weill-dyschondrosteosis in their left-hand radiography, namely triangularization of the distal radial epiphysis, pyramidalization of the distal carpal row, or lucency of the distal ulnar border of the radius. Our observations suggest that it is rational to limit SHOX mutation screening to children with an extremities-trunk ratio less than 1.95 + 1/2 height (m) and to add a critical judgment of the hand radiography. PMID- 14557472 TI - Insulin resistance, hemostatic factors, and hormone interactions in pre- and perimenopausal women: SWAN. AB - We evaluated the association of hemostatic factors with insulin resistance in relation to reproductive hormones including FSH, estradiol, testosterone, and SHBG. SHBG was used to calculate the free estradiol index and free androgen index. We studied 3,200 women, aged 42-52 yr, in the Study of Women's Health Across the Nation, a prospective multiethnic study of the menopausal transition. We measured the hemostatic factors, fibrinogen, factor VIIc, tissue plasminogen activator (t-PA), and plasminogen activator inhibitor type 1 (PAI-1), as well as glucose and insulin to calculate insulin resistance. After adjustment for body mass index, site, and ethnicity, SHBG was correlated with PAI-1 (partial r = 0.30) and t-PA (partial r = -0.12). Although testosterone was associated with t PA (partial r = 0.13) and PAI-1 (partial r = 0.07), free androgen index was strongly correlated with t-PA (partial r = 0.18) and PAI-1 (partial r = 0.26). SHBG modified the association of hemostatic factors with insulin resistance. Women with greater insulin resistance had lower SHBG and higher PAI-1. Estrogen measures were not associated with insulin resistance. The influence of sex hormones on hemostatic factors and insulin resistance is poorly understood. SHBG, which influences the amount of bioavailable hormone, significantly modified the association of PAI-1 and t-PA with insulin resistance. The longitudinal Study of Women's Health Across the Nation will help us discern whether this interaction contributes to heart disease and diabetes among postmenopausal women. PMID- 14557471 TI - The pathophysiology of diabetes involves a defective amplification of the late phase insulin response to glucose by glucose-dependent insulinotropic polypeptide regardless of etiology and phenotype. AB - The effect of the insulinotropic incretin hormone, glucagon-like peptide-1 (GLP 1), is preserved in typical middle-aged, obese, insulin-resistant type 2 diabetic patients, whereas a defective amplification of the so-called late-phase plasma insulin response (20-120 min) to glucose by the other incretin hormone, glucose dependent insulinotropic polypeptide (GIP), is seen in these patients. The aim of the present investigation was to evaluate plasma insulin and C-peptide responses to GLP-1 and GIP in five groups of diabetic patients with etiology and phenotype distinct from the obese type 2 diabetic patients. We studied (six in each group): 1) patients with diabetes mellitus secondary to chronic pancreatitis; 2) lean type 2 diabetic patients (body mass index < 25 kg/m(2)); 3) patients with latent autoimmune diabetes in adults; 4) diabetic patients with mutations in the HNF 1alpha gene [maturity-onset diabetes of the young (MODY)3]; and 5) newly diagnosed type 1 diabetic patients. All participants underwent three hyperglycemic clamps (2 h, 15 mM) with continuous infusion of saline, 1 pmol GLP 1 (7-36)amide/kg body weight.min or 4 pmol GIP pmol/kg body weight.min. The early phase (0-20 min) plasma insulin response tended to be enhanced by both GIP and GLP-1, compared with glucose alone, in all five groups. In contrast, the late phase (20-120 min) plasma insulin response to GIP was attenuated, compared with the plasma insulin response to GLP-1, in all five groups. Significantly higher glucose infusion rates were required during the late phase of the GLP-1 stimulation, compared with the GIP stimulation. In conclusion, lack of GIP amplification of the late-phase plasma insulin response to glucose seems to be a consequence of diabetes mellitus, characterizing most, if not all, forms of diabetes. PMID- 14557473 TI - Intronic single nucleotide polymorphisms in the RET protooncogene are associated with a subset of apparently sporadic pheochromocytoma and may modulate age of onset. AB - Approximately 75% of pheochromocytomas are sporadic. Germline mutations in RET, VHL, SDHB, and SDHD have been shown to cause the 25% that are hereditary. Germline high penetrance gain-of-function RET mutations cause multiple endocrine neoplasia type 2, of which medullary thyroid carcinoma (MTC) and pheochromocytoma are components, whereas loss-of-function mutations cause Hirschprung disease (HSCR). A low-penetrance founder locus, in linkage disequilibrium with a RET ancestral haplotype comprising specific alleles at three intron (IVS) 1 single nucleotide polymorphisms (SNPs) (haplotype 0) and SNP A45A, predisposes to the majority of isolated HSCR. A different low-penetrance locus, in linkage disequilibrium with IVS 1 haplotype 2 and SNP S836S, was associated with a subset of sporadic MTC. We, therefore, sought to determine whether RET might also be a low-penetrance gene for apparently sporadic pheochromocytoma. We analyzed 104 pheochromocytoma cases without germline mutations in RET, VHL, SDHD, and SDHB for their status at A45, S836, three IVS 1 SNPs, and a novel upstream insertion/deletion variant. Pheochromocytoma cases were not associated with either A45A or S836S, but we found that cases were associated with haplotype 0 (P = 0.032). However, unlike HSCR, this pheochromocytoma-associated haplotype 0 was not associated with A45A. Taken together with the strengthening of association with the addition of the 5' insertion/deletion variant data (P = 0.016), our observations suggest the presence of a low-penetrance pheochromocytoma susceptibility locus in a region upstream of the putative loci for HSCR and apparently sporadic MTC. PMID- 14557474 TI - Modified quantitative insulin sensitivity check index is better correlated to hyperinsulinemic glucose clamp than other fasting-based index of insulin sensitivity in different insulin-resistant states. AB - Fasting-based index estimates of insulin sensitivity were compared with euglycemic hyperinsulinemic clamp (IS clamp) measurements in 148 subjects: normal controls (n = 46), and obese (n = 12), polycystic ovary syndrome (n = 16), first degree relatives of type 2 diabetic (n = 17), impaired glucose tolerance (n = 28), and type 2 diabetic (n = 29) patients. The fasting-based indexes tested included log homeostasis model assessment (HOMA), the quantitative insulin sensitivity check index (QUICKI), the revised QUICKI, and a new revised QUICKI using fasting plasma glycerol. In the population studied, at 40 mU/m(2).min (n = 30) revised QUICKI (r = 0.86; P < 0.0001) and QUICKI-glycerol (r = 0.87; P < 0.0001) gave higher correlations with the IS clamp than QUICKI and log HOMA (r = 0.78 and r = -0.78; P < 0.001). For subjects tested at 75 mU/m(2).min (n = 118), comparable correlations were found for all indexes (r > 0.80; P < 0.0001). When studied in subgroups, revised QUICKI and QUICKI-glycerol give significantly higher correlations with the IS clamp than other indexes for lean control subjects studied at 40mU/m(2).min and impaired glucose tolerance subjects. We confirmed, in a large patient population with a wide range of insulin sensitivities, that no single test is superior in all groups of patients. However, QUICKI and revised QUICKI are good indexes that offer correlations similar to or higher than values obtained with log HOMA. Such indexes are simple tools to estimate insulin sensitivity appropriate for epidemiological studies. PMID- 14557475 TI - Body fat distribution and cortisol metabolism in healthy men: enhanced 5beta reductase and lower cortisol/cortisone metabolite ratios in men with fatty liver. AB - In Cushing's syndrome, cortisol causes fat accumulation in specific sites most likely to be associated with insulin resistance, notably in omental adipose and also perhaps in the liver. In idiopathic obesity, cortisol-metabolizing enzymes may play a key role in determining body fat distribution. Increased regeneration of cortisol from cortisone within adipose by 11beta-hydroxysteroid dehydrogenase (HSD) type 1 (11HSD1) has been proposed to cause visceral fat accumulation, whereas decreased hepatic 11HSD1 may protect the liver from glucocorticoid excess. Increased inactivation of cortisol by 5alpha- and 5beta-reductases in the liver may drive compensatory activation of the hypothalamic-pituitary-adrenal axis, hence increasing adrenal androgens and 'android' central obesity. This study aimed to examine relationships between these enzymes and detailed measurements of body fat distribution. Twenty-five healthy men (age, 22-57 yr; body mass index, 20.6-35.6 kg/m(2)) were recruited from occupational health services. Body composition was assessed by anthropometric measurements, bioimpedance, and cross-sectional abdominal magnetic resonance imaging scans. Liver fat content was assessed by magnetic resonance imaging spectroscopy. Insulin sensitivity was measured in a euglycemic hyperinsulinemic clamp. Cortisol metabolites were measured in a 24-h urine sample by gas chromatography-mass spectrometry. In vivo hepatic 11HSD1 activity was measured by generation of plasma cortisol after an oral dose of cortisone. In vitro 11HSD1 activity and mRNA were measured in 18 subjects who consented to provide abdominal sc adipose biopsies. Indices of obesity (body mass index, whole-body percentage fat, waist/hip ratio) were associated with higher urinary excretion of 5alpha- and 5beta-reduced cortisol metabolites (for percentage fat, P < 0.05 and P < 0.01, respectively) and increased adipose 11HSD1 activity (P < 0.05). Liver fat accumulation was associated with a selective increase in urinary excretion of 5beta-reduced cortisol and cortisone metabolites (P < 0.01) and a lower ratio of cortisol/cortisone metabolites in urine (P < 0.001) but no difference in in vivo cortisone-to-cortisol conversion or in vitro adipose 11HSD1. Higher excretion of 5beta-reduced cortisol metabolites was independently associated with insulin resistance and hypertriglyceridemia. Lower conversion of cortisone to cortisol was associated with lower fasting plasma cortisol (P < 0.01). However, visceral adipose fat mass was not associated with indices of cortisol metabolism; indeed, after adjusting for the effects of whole-body and liver fat, increased visceral fat was associated with lower cortisol metabolite excretion. We conclude that alterations in 11HSD1 and hepatic 5alpha-reductase activity are associated with generalized, rather than central, obesity in humans. Activation of 5beta reductase in men with fat accumulation in the liver may confound the interpretation of cortisol metabolite excretion when liver fat content is unknown, and may contribute to altered bile acid and cholesterol metabolism in nonalcoholic steatohepatitis. PMID- 14557477 TI - Expression of interleukin-6 and its effects on growth of HP75 human pituitary tumor cells. AB - The role of IL-6 in the pathogenesis of pituitary adenomas is unclear, as tumor biology is difficult to study in primary culture. We have shown here that the human pituitary cell line HP75 synthesizes IL-6 mRNA and expresses and secretes IL-6 (6167 +/- 56 pg/ml/72 h for 30,000 cells). IL-6 receptor (IL-6R) mRNA was identified by in situ hybridization and RT-PCR. Exogenous IL-6 in low dose (1 ng/ml) stimulated, whereas higher doses (100 ng/ml) inhibited, growth. This diverse effect occurs in other cell types as a result of receptor down regulation. Cell growth was inhibited by IL-6-blocking antibody (76 +/- 6.5% inhibition; P < 0.0001). This demonstrates that IL-6 is an important growth regulator in HP75 cells, having an autocrine growth stimulatory effect under basal conditions. IL-1alpha and dibutyryl cAMP stimulated and dexamethasone inhibited IL-6 secretion; however, bacterial lipopolysaccharide, forskolin, and cholera toxin had no effect. This implies that there is a defect in the control of IL-6 secretion. Soluble IL-6R was not detected, but soluble gp130 receptor was present in the conditioned medium. Stimulation of cleavage of soluble IL-6R from the membrane-bound IL-6R could not be induced by phorbol ester or dexamethasone. Whether IL-6 has a similar effect in human pituitary adenomas requires further investigation. PMID- 14557476 TI - Germline succinate dehydrogenase subunit D mutation segregating with familial non RET C cell hyperplasia. AB - C cell hyperplasia is associated with medullary carcinoma of the thyroid in the inherited MEN2 syndromes, in which the great majority of cases have been shown to be due to a mutation in the RET oncogene. We report a study of a family with C cell hyperplasia and hypercalcitoninemia in which no cases of medullary carcinoma have yet occurred and which lacked an identifiable causative RET mutation. Four of the family members showed hypercalcitoninemia, and marked C cell hyperplasia was present in each of the three in whom thyroidectomy has been performed. We investigated the possible involvement of the SDHD gene, because somatic and germline mutations in this gene have been found in a variety of tumors of neural crest-derived tissue. A germline mutation in exon 2 of the SDHD gene (c149 A-G, His 50 Arg) was found in six members of the family; all the four available members with hypercalcitoninemia possessed the mutation. One of the five available members without hypercalcitoninemia, an 18-yr-old female, also showed the mutation. We conclude that we have identified a new syndrome, characterized by familial non-RET C cell hyperplasia. Our studies suggest that a mutation in SDHD may be causative. These observations have implications for apparently incidental cases of hypercalcitoninemia or C cell hyperplasia. PMID- 14557478 TI - Intercellular adhesion molecule 1 gene polymorphisms in Graves' disease. AB - It was recently suggested that genetic factors could play a major role in the development of Graves' disease (GD). The aim of the present study was to evaluate the frequency of the c.721G-->A polymorphism and the c.1405A-->G polymorphism of the intercellular adhesion molecule 1 (ICAM-1) gene in subjects with GD compared with that in healthy controls, because ICAM-1 was found to play a key role in lymphocyte infiltration into the thyroid gland and the concentration of the soluble form of ICAM-1 correlates significantly with the clinical activity and treatment status in GD. We have analyzed the association of ICAM-1 polymorphisms with the age at onset of GD and the presence of ophthalmopathy. In a group of 235 patients with GD and 211 healthy controls we have shown that polymorphism at position c.721G-->A is associated with an earlier age of GD onset and that the c.1405A-->G polymorphism of the ICAM-1 gene could predispose to Graves' ophthalmopathy. This suggests that G241R and K469E amino acid substitutions in the ICAM-1 molecule could influence the intensity/duration of the autoimmunity process and the infiltration of orbital tissues. It could be speculated that therapy that modulates ICAM-1 function may delay the onset and/or prolong the remission and/or have an influence on clinical manifestations of GD. PMID- 14557479 TI - Oxidative stress does not modulate metabolic rate or skeletal muscle sympathetic activity with primary aging in adult humans. AB - Support of resting metabolic rate (RMR) by the beta-adrenergic receptors of the sympathetic nervous system is attenuated with age and contributes to declines in RMR. This may be mediated by an age-associated increase in oxidative stress that can suppress beta-adrenergic responsiveness and/or modulate sympathetic activity. To address these issues, RMR was determined in 12 young (23 +/- 1 yr, mean +/- SE) and 21 older (68 +/- 3 yr) adults before and during systemic infusion of ascorbic acid [bolus, 0.06 g/kg fat free mass (FFM); drip, 0.02]. Ascorbic acid increased plasma concentrations similarly in young (72 +/- 5 to 1107 +/- 114 micro mol/liter) and older (70 +/- 6 to 1022 +/- 63 micro mol/liter) adults, and reduced (P = 0.001) plasma concentrations of isoprostanes (young, -82.8 +/- 47; older, -107 +/- 29 pg/ml). Baseline RMR(FFM) was lower (5719 +/- 215 vs. 6703 +/- 328 kJ/d; P = 0.001) and muscle sympathetic nerve activity (MSNA) was greater (MSNA, 28 +/- 2 vs. 23 +/- 3 bursts/min; P < 0.05) in older compared with young. However, neither RMR(FFM) (young, +117 +/- 63; older, +163 +/- 48 kJ/d; P = 0.14) or MSNA (young, 0 +/- 2; older, -1 +/- 1 bursts/min; P = 0.71) changed in either age group during ascorbic acid infusion compared with saline control. These results indicate that increased oxidative stress: 1) is not a mechanism contributing to decreases in RMR with primary aging; and 2) does not modulate MSNA in healthy adult humans. PMID- 14557480 TI - Bound leptin and sympathetic outflow in nonobese men. AB - Leptin exists in a free form and a receptor-bound form. Protein-bound rather than free leptin levels may be associated with regulation of muscle sympathetic nerve activity (MSNA). We determined MSNA and bound leptin concentrations in 25 men [age, 29 +/- 6 yr, body mass index (BMI), 24 +/- 3 kg/m(2)]. Baroreflex sensitivity was measured using phenylephrine and nitroprusside infusions. We measured bound leptin in patients with central (multiple system atrophy, n = 8; age, 59 +/- 8 yr; BMI, 23 +/- 2 kg/m(2)) and peripheral autonomic failure (pure autonomic failure, n = 4; age, 71 +/- 10 yr; BMI, 25 +/- 3 kg/m(2)). MSNA was correlated with protein-bound leptin concentrations (r(2) = 0.35; P < 0.01) but not with free leptin levels (r(2) = 0.09). MSNA at baseline was 15 +/- 2 bursts x minutes(-1) in subjects with lower and 24 +/- 3 bursts x minutes(-1) in subjects with higher bound leptin concentrations (P < 0.05). Blood pressure as well as baroreflex regulation of heart rate and MSNA was similar in both groups. Phenylephrine and nitroprusside responses were similar. Patients with multiple system atrophy and autonomic failure featured similar bound leptin levels. We conclude that protein-bound rather than free leptin levels are correlated with basal sympathetic outflow in normotensive, nonobese men. This relationship cannot be explained by a direct central nervous effect of protein-bound leptin. Instead, protein-bound leptin may increase sympathetic vasomotor tone indirectly via a baroreflex mechanism. PMID- 14557481 TI - Laminin isoforms in fetal and adult human adrenal cortex. AB - Laminin has been proposed to influence the function of human adrenal cortex. We have studied the distribution of laminin (Ln) chains using immunofluorescence in human fetal and adult adrenal cortex. In the fetal gland Ln alpha2- and alpha5 chains were weakly expressed in the definitive zone, whereas Ln alpha4-, beta1-, and gamma1-chains occurred around vessels. In the adult gland, Ln alpha2-, alpha5 , and gamma1-chains were found in epithelial basement membranes (BM) in all cortical zones, Ln alpha4-chain in vessels, Ln beta1-chain in outer zone, and Ln beta2-chain in the two inner zones of the cortex, respectively. Among the integrins in adult gland, integrin alpha(3)-subunit was confined to basal surfaces of cortical cells, alpha(6) to vessels, alpha(1) to the stroma, and alpha(2) diffusely to epithelial cells. Lutheran glycoprotein and dystroglycan occurred in the fetal gland diffusely in the definitive zone and throughout the epithelium in the adult. The isoform composition of BM of the adult adrenal gland is distinct, with Ln-2 and -10 in BM of the outer zone and Ln-4 and -11 in BM of the two inner zones. The results suggest that integrin alpha(3)beta(1) and Lutheran are candidate receptors for Ln-10 and -11, whereas dystroglycan probably binds Ln-2 and -4. PMID- 14557482 TI - The expression of Smads in human endometrium and regulation and induction in endometrial epithelial and stromal cells by transforming growth factor-beta. AB - Human endometrium expresses TGF-beta and TGF-beta receptors where they regulate several endometrial biological activities implicated in embryo implantation, irregular bleeding, endometriosis, and cancer. In the present study, we determined the expression of Smads, intracellular signals that mediate TGF-beta receptors signals from the cell surface to the nucleus, in the endometrium as well as isolated endometrial epithelial (EEC) and stromal (ESC) cells. We also determined whether TGF-beta regulates the expression Smads and activates Smad3 in these cells and endometrial surface epithelial cell line (HES). Using semiquantitative RT-PCR, Western blot analysis, and immunohistochemistry, we found that endometrium, EEC, ESC, and HES express Smad3, -4, and -7 mRNA and protein and contain phosphorylated Smad3 (pSmad3). Smads and pSmad3 were localized in the epithelial and stromal cells with cytoplasmic/nuclear localization. TGF-beta in a dose- and time-dependent manner increased the expression of Smads mRNA and protein, the rate of pSmad3 activation, and Smad3 translocation into the nucleus in ESC and HES. The effect of TGF-beta on pSmad3 induction was, in part, abrogated by the pretreatment of HES and ESC with TGF beta type II receptor antisense oligonucleotides. We conclude that human endometrium expresses the necessary components of Smad signaling pathway, whose expression and induction in endometrial epithelial and stromal cells are regulated by TGF-beta. PMID- 14557483 TI - Correlation between the loss of thyroglobulin iodination and the expression of thyroid-specific proteins involved in iodine metabolism in thyroid carcinomas. AB - Progress in biotechnology has provided useful tools for tracing proteins involved in thyroid hormone synthesis in vivo. Mono- or polyclonal antibodies are now available to detect on histological sections the Na(+)/I(-) symporter (NIS) at the basolateral pole of the cell, the putative iodide channel (pendrin) at the apical plasma membrane, thyroperoxidase (TPO), and members of the NADPH-oxidase family, thyroid oxidase 1 and 2 (ThOXs), part of the H(2)O(2)-generating system. The aim of this study was to correlate thyroglobulin (Tg) iodination with the presence of these proteins. Tg, T(4)-containing Tg, NIS, pendrin, TPO, ThOXs, and TSH receptor (TSHr) were detected by immunohistochemistry on tissue sections of normal thyroids and various benign and malignant thyroid disorders. Tg was present in all cases. T(4)-containing Tg was found in the adenomas, except in Hurthle cell adenomas. It was never detected in carcinomas. NIS was reduced in all types of carcinomas, whereas it was detected in noncancerous tissues. Pendrin was not expressed in carcinomas, except in follicular carcinomas, where weak staining persisted. TPO expression was present in insular, follicular carcinomas and in follicular variants of papillary carcinomas, but in a reduced percentage of cells. It was below the level of detection in papillary carcinomas. The H(2)O(2)-generating system, ThOXs, was found in all carcinomas and was even increased in papillary carcinomas. Its staining was apical in normal thyroids, whereas it was cytoplasmic in carcinomas. The TSHr was expressed in all cases, but the intensity of the staining was decreased in insular carcinomas. In conclusion, our work shows that all types of carcinomas lose the capacity to synthesize T(4)-rich, iodinated Tg. In follicular carcinomas, this might be due to a defect in iodide transport at the basolateral pole of the cell. In papillary carcinomas, this defect seems to be coupled to an altered apical transport of iodide and probably TPO activity. The TSHr persists in virtually all cases. PMID- 14557484 TI - Sertoli cell proliferation during prepubertal development in the rhesus monkey (Macaca mulatta) is maximal during infancy when gonadotropin secretion is robust. AB - Although a marked pubertal increase in Sertoli cell number is a hallmark of testicular development in the rhesus monkey, the ontogeny of this somatic cell type before puberty is less clear. To clarify this issue, groups (n = 4) of neonate (1-2 d old), infant (4-5 months old), juvenile (14-17 months old), and adult male rhesus monkeys were injected with 5-bromo-2'-deoxyuridine (BrdU) 2 h before castration. Tissue was fixed in Bouin's fluid, and the percentage of BrdU labeled Sertoli cells at each developmental stage was calculated. In addition to the labeling index, Sertoli cell number per testis for the neonate and infant groups was enumerated using standard histomorphometry and compared with that previously reported by this laboratory for juvenile and adult rhesus monkeys. The number of Sertoli cells per testis in infants (156 +/- 49 x 10(6), mean +/- SD) was 4-fold greater than that in neonates (42 +/- 12 x 10(6)). The previously established value for this parameter in juvenile monkeys was 286 +/- 121 x 10(6). Incorporation of BrdU into nuclei of Sertoli cells indicated that these cells were mitotically active at all three stages of prepubertal development. The labeling index in the neonate and infant groups (1.33% in both cases), however, was significantly greater than that in juveniles (0.25%). From the foregoing results, we conclude that Sertoli cell proliferation during prepubertal development in the rhesus monkey occurs predominantly during infancy, when gonadotropin secretion is elevated, and to a lesser extent during the juvenile phase of development, when circulating gonadotropin concentrations are undetectable. PMID- 14557485 TI - Enhanced expression of nicotinamide N-methyltransferase in human papillary thyroid carcinoma cells. AB - To gain an understanding of the molecular pathogenesis of thyroid cancer, we used DNA microarray to study the expression profiles of 10 different human thyroid carcinoma cell lines. These included papillary lines BHP 2-7, BHP 7-13, BHP 10-3, BHP 18-21, NPA 87, and TPC1; anaplastic lines ARO 81-1 and DRO 90-1; follicular line WRO 82-1; and medullary line HRO 85-1. Among the genes with increased expression in the cancer cell lines, a gene coding for nicotinamide N methyltransferase (NNMT) was identified for being highly expressed only in the papillary cell lines. NNMT catalyzes N-methylation of nicotinamide and other structurally related compounds and is highly expressed in the human liver. The results were further confirmed by semiquantitative RT-PCR and Northern blot analysis. NNMT catalytic activities were determined in all of the cells described above and in additional cell lines. Significantly higher NNMT enzyme activities were detected in eight of 10 of the papillary lines and three of six of the follicular cell lines tested. Normal thyroid tissue, thyroid primary cultures, anaplastic cancer cells, and medullary cancer cells showed no or low enzyme activity. Immunohistochemical staining for NNMT of human thyroid specimens showed strong and abundant cytoplasmic reactions in the sections of papillary carcinomas, and weak or scanty reaction in the normal thyroid tissues. These results indicate that NNMT is a potential biomarker for papillary thyroid carcinoma. PMID- 14557486 TI - Reduced expression of immunoreactive beta2-adrenergic receptor protein in human myometrium with labor. AB - A considerable body of evidence exists suggesting that the beta(2)-adrenergic receptor (beta(2)-AR) mediates uterine relaxation. However, little information exists on the localization, distribution, or expression of beta(2)-ARs in the human myometrium during the nonpregnant to labor transition. We have used immunochemical methods to investigate beta(2)-AR localization and expression in the nonpregnant, term pregnant, and term parturient uterus. Myometrial biopsies were obtained from 1) nonpregnant, menstruating women undergoing hysterectomy; 2) singleton term pregnant women undergoing elective cesarean section before the onset of labor; or 3) singleton term pregnant women undergoing emergency cesarean section after spontaneous labor. Tissues were processed for immunohistochemistry, immunofluorescence, and Western blotting and a primary polyclonal antibody specific to the human beta(2)-AR to identify immunoreactive myometrial beta(2) AR. Protein levels were subsequently quantified by densitometry relative to rat brain protein. Immunohistochemistry and immunofluorescence demonstrated the presence of beta(2)-AR predominantly at the plasma membrane and also in the cytosol of myometrial cells. A 2-fold decrease in protein levels of the beta(2) AR was apparent in the myometrium of labor compared with that of nonpregnant and pregnant nonlaboring women (P < 0.05). These results demonstrate that down regulation of beta(2)-AR protein with labor may constitute a contributory mechanism by which uterine quiescence is removed at term. PMID- 14557487 TI - Expression of betaglycan, an inhibin coreceptor, in normal human ovaries and ovarian sex cord-stromal tumors and its regulation in cultured human granulosa luteal cells. AB - Activins and inhibins are often antagonistic in the regulation of ovarian function. TGFbeta type III receptor, betaglycan, has been identified as a coreceptor to enhance the binding of inhibins to activin type II receptor and thus to prevent the binding of activins to their receptor. In this study we characterized the expression and regulation pattern of betaglycan gene in normal ovaries and sex cord-stromal tumors and in cultured human granulosa-luteal cells from women undergoing in vitro fertilization. Expression of betaglycan mRNA was detected by RT-PCR or Northern blotting in normal ovarian granulosa, thecal, and stroma cells as well as in granulosa-luteal cells. Immunohistochemical analysis revealed positive staining for betaglycan in antral and preovulatory follicular granulosa and thecal cells and in corpora lutea of normal ovaries. Furthermore, betaglycan expression was detected in the vast majority of granulosa cell tumors, thecomas, and fibromas, with weaker staining in granulosa cell tumors compared with fibrothecomas. In cultured granulosa-luteal cells, FSH and LH treatment increased dose-dependently the accumulation of betaglycan mRNA, as did the protein kinase A activator dibutyryl cAMP and the protein kinase C inhibitor staurosporine. In contrast, the protein kinase C activator 12-O-tetradecanoyl phorbol 13-acetate had no significant effect on betaglycan mRNA levels. Treatment with prostaglandin E(2) and with its receptor EP2 subtype agonist butaprost increased betaglycan mRNA accumulation and progesterone secretion dose- and time dependently. In summary, betaglycan gene is expressed in normal human ovarian steroidogenic cells and sex cord-stromal ovarian tumors. The accumulation of its mRNA in cultured granulosa-luteal cells is up-regulated by gonadotropins and prostaglandin E(2), probably via the protein kinase A pathway. The specific expression and regulation pattern of betaglycan gene may be related to the functional antagonism of inhibins to activin signal transduction in human ovaries. PMID- 14557488 TI - Enhanced polyadenosine diphosphate-ribosylation in gonadotropin-releasing hormone agonist-treated uterine leiomyoma. AB - This study aimed to examine the activation of poly(ADP-ribose) polymerase (PARP) and the accumulation of its end product, poly(ADP-ribose) (PAR), in uterine leiomyoma specimens obtained from 25 patients receiving Leuplin depot [leuprorelin acetate, depot (LA)] treatment and 46 control patients and explore their correlation with tumor shrinkage and degeneration caused by the therapy. Immunoblotting analysis showed that specimens from LA-treated patients had higher PARP expression. The numbers of both PARP- and PAR-immunolabeled cells were higher in leiomyoma with LA treatment. This was correlated with the clinical response of LA therapy that LA induced more leiomyoma degeneration. The analysis of power Doppler sonography indicated a progressive decrease in blood supply to tumor following LA treatment. In vitro experiments using primarily cultured leiomyoma cells exhibited that the deprivation of serum or ovarian hormones or LA directly failed to induce PARP and PAR production. Our results suggested that reduced blood flow and subsequent ischemic damages in leiomyoma could be responsible for PARP overexpression and PAR accumulation, clinical response, and tumor degeneration caused by LA treatment. PMID- 14557489 TI - Regulation of PTEN (phosphatase and tensin homolog deleted on chromosome 10) expression by estradiol and progesterone in human endometrium. AB - PTEN (phosphatase and tensin homolog deleted on chromosome 10) is a tumor suppressor gene, mutated frequently in a variety of human tumors. PTEN regulates cell growth, apoptosis, and proliferation. Phosphorylation in PTEN tail causes its inactivation and decreases its degradation. There is little known about the regulation of PTEN by ovarian steroids. We hypothesized that PTEN expression in human endometrium is variable throughout the menstrual cycle and early pregnancy, and that ovarian steroids regulate PTEN expression because PTEN is critical in many steroid-sensitive tissues such as endometrium, prostate, and breast. In the present study, we have observed a direct regulation of PTEN by ovarian steroids. Estradiol increased PTEN phosphorylation at 5-15 min. After 24-h treatment, progesterone induced a significant increase in PTEN protein levels, assessed by Western blot. Furthermore, we evaluated for the first time a comparison between menstrual cycle and early pregnancy, immunohistochemically. Endometrial PTEN expression revealed temporal and spatial changes throughout the menstrual cycle and during early pregnancy. We conclude that estradiol may down-regulate PTEN activity by increasing its phosphorylation, but progesterone is likely to regulate the PTEN pool by decreasing its phosphorylation and increasing its protein level. Presented data, therefore, suggest that ovarian steroids regulate the endometrial PTEN pool. We propose that PTEN might be one of the signaling proteins that estrogen and progesterone are acting to affect endometrial cell proliferation and/or apoptosis. PMID- 14557490 TI - ONYX-015 enhances radiation-induced death of human anaplastic thyroid carcinoma cells. AB - ONYX-015 is a genetically modified adenovirus with a deletion of the E1B early gene and therefore is designed to replicate preferentially in p53-mutated cells causing their death. We previously demonstrated that the ONYX-015 virus kills anaplastic thyroid carcinoma (ATC) cells and enhances the antineoplastic effects of doxorubicin and paclitaxel. Here we report that ONYX-015 increased the cytopathic effect of radiotherapy in three ATC cell lines. In fact, ONYX-015 and radiation induced a significant cytopathic effect on ATC cells. DNA fragmentation analysis showed that ATC ONYX-015-treated cells were very sensitive to radiation induced apoptosis. In addition, low doses of ONYX-015 associated with a single radiation dose of 10 Gy delayed the growth of a xenograft tumor induced by ARO cells in athymic mice. Our results suggest that the combination of ONYX-015 and radiotherapy should be considered for experimental trials in patients with anaplastic thyroid carcinoma. PMID- 14557491 TI - Late gestation increase in 11beta-hydroxysteroid dehydrogenase 1 expression in human fetal membranes: a novel intrauterine source of cortisol. AB - Late human gestation is associated with an increase in the concentration of cortisol (F) in the fetal circulation and amniotic fluid. It had been assumed that most of the F measured in the amniotic fluid came from the fetal adrenal gland. However, local production of F can also occur in human intrauterine tissues from inactive cortisone under the influence of the enzyme 11beta hydroxysteroid dehydrogenase (11beta-HSD) type 1. Recent studies have shown that 11beta-HSD 1 activity is up-regulated by prostaglandins (PG) E2 and F2alpha, hormones that are produced in the fetal membranes (FM) at term. In the present study, we hypothesized that 11beta-HSD 1 expression would increase in FM during pregnancy and at labor, creating the potential for local increase in F production at term. We examined 11beta-HSD 1 expression in placenta and FM obtained during normal pregnancy from nonlaboring women [26-28 wk (n = 3); 29-30 wk (n = 3); 32 33 wk (n = 3); 35-36 wk (n = 3)] and from uncomplicated term pregnancies after elective cesarean section (n = 6). 11beta-HSD 1 expression was also examined in amnion and chorionic tissues in relation to term labor (n = 12). Immunohistochemistry and Western blot analysis were used to examine 11beta-HSD 1 localization and expression. 11beta-HSD 1 activity was also measured in microsomal fractions prepared from whole fetal membranes. At term, immunoreactive 11beta-HSD 1 expression was localized predominantly to the chorion trophoblast cells, attached decidua, and amnion epithelial cells. 11beta-HSD 1 expression in FM increased with gestational age and reflected increased enzyme reductase activity. No change in 11beta-HSD 1 expression was found in placental tissue from the same patients. There was a significant increase in 11beta-HSD 1 expression in amnion but not in chorion with the onset of labor. We suggest that increases in 11beta-HSD 1 expression/activity by intrauterine membranes during late gestation may result in increased potential for a local increase in F production and that FM should be considered as an extraadrenal source of F during late gestation. This local F production may be involved in different pathways contributing to the regulation of parturition. PMID- 14557492 TI - Association of a rare thyroglobulin gene microsatellite variant with autoimmune thyroid disease. AB - Genetic and environmental factors contribute to the development of Graves' disease and Hashimoto's thyroiditis. These diseases, although clinically distinct, share many immunological and histological features. Susceptibility genes for autoimmune thyroid disease (AITD) have been investigated, although only the human leukocyte antigen and cytotoxic T lymphocyte-associated antigen-4 gene regions have been consistently associated with disease. Recent data, however, have shown linkage and association of chromosome 8q24 (containing the thyroglobulin gene) to AITD. Therefore, we performed a case-control association study on patients with AITD and controls using previously associated markers (D8S284 and Tgms2). No differences in allele frequencies were observed between AITD cases and controls for D8S284. Compared with the three common alleles (frequencies >10%), the rare alleles of Tgms2 were increased (chi(2)= 10.6; P = 0.001) at Tgms2. This group included the 336-bp allele (increased in cases vs. controls: chi(2)= 24.97; P < 0.001), which has previously been reported to be associated with AITD. The rarity of this allele in the United Kingdom, however, precluded analysis in our family dataset. Although these findings may represent a random chance event, in view of previous reports of linkage and association of this gene region to AITD, this may be an example of a rare causal variant of a complex disease. PMID- 14557493 TI - Human placental growth hormone, insulin-like growth factor I and -II, and insulin requirements during pregnancy in type 1 diabetes. AB - Human placental GH (hPGH) replaces pituitary GH during pregnancy. hPGH is correlated to serum IGF-I in normal pregnancies and in pregnancies complicated by fetoplacental disorders. In gestational diabetes and type 2 diabetes no correlation between hPGH and IGF-I has been found. The relationship between hPGH and IGF-I in type 1 diabetes mellitus has not been investigated thoroughly. Furthermore, hPGH may be involved in the development of insulin resistance during pregnancy. In this prospective, longitudinal study, 51 type 1 diabetic subjects were followed with repeated blood sampling during pregnancy (median, 14 blood samples/subject; range, 8-26). Maternal concentrations of serum hPGH, IGF-I, and IGF-II were measured and compared with insulin requirements and birth characteristics. hPGH was detected from as early as 6 wk gestation. In all subjects, a rise in serum hPGH was observed during pregnancy, and the rise between wk 16 and 25 was correlated to the rise between wk 26 and 35 (P < 0.001). From wk 26 onward, the increase in hPGH values was significantly correlated to the birth weight, expressed as a z-score (r(s) = 0.54; P < 0.001), as were the absolute hPGH values. Also, a positive influence of hPGH on placental weight was found. Serum IGF-I values decreased significantly from the first to the second trimester (P 30 precise sites of ubiquitination. Major classes of identified ubiquitinated proteins include ER resident membrane proteins, plasma membrane-localized permeases, receptors, and enzymes, and surprisingly, components of the actin cytoskeleton. By determining the differential abundance of ubiquitinated proteins in yeast mutated for NPL4 and UBC7, which are major components of ER-associated degradation (ERAD), we furthermore were able to classify 83 of these identified ubiquitinated membrane proteins as potential endogenous substrates of the ERAD pathway. These substrates are highly enriched for proteins that localize to or transit through the ER. Interestingly, we also identified novel membrane-bound transcription factors that may be subject to ubiquitin/proteasome-mediated cleavage and activation at the ER membrane. PMID- 14557539 TI - Elevated gene expression levels distinguish human from non-human primate brains. AB - Little is known about how the human brain differs from that of our closest relatives. To investigate the genetic basis of human specializations in brain organization and cognition, we compared gene expression profiles for the cerebral cortex of humans, chimpanzees, and rhesus macaques by using several independent techniques. We identified 169 genes that exhibited expression differences between human and chimpanzee cortex, and 91 were ascribed to the human lineage by using macaques as an outgroup. Surprisingly, most differences between the brains of humans and non-human primates involved up-regulation, with approximately 90% of the genes being more highly expressed in humans. By contrast, in the comparison of human and chimpanzee heart and liver, the numbers of up- and down-regulated genes were nearly identical. Our results indicate that the human brain displays a distinctive pattern of gene expression relative to non-human primates, with higher expression levels for many genes belonging to a wide variety of functional classes. The increased expression of these genes could provide the basis for extensive modifications of cerebral physiology and function in humans and suggests that the human brain is characterized by elevated levels of neuronal activity. PMID- 14557541 TI - Coloration strategies in peacock feathers. AB - We report the mechanism of color production in peacock feathers. We find that the cortex in differently colored barbules, which contains a 2D photonic-crystal structure, is responsible for coloration. Simulations reveal that the photonic crystal structure possesses a partial photonic bandgap along the direction normal to the cortex surface, for frequencies within which light is strongly reflected. Coloration strategies in peacock feathers are very ingenious and simple: controlling the lattice constant and the number of periods in the photonic crystal structure. Varying the lattice constant produces diversified colors. The reduction of the number of periods brings additional colors, causing mixed coloration. PMID- 14557542 TI - Worker policing without genetic conflicts in a clonal ant. AB - In group-living animals, mutual policing to suppress reproduction is an important mechanism in the resolution of conflict between selfish group members and the group as a whole. In societies of bees, ants, and wasps, policing against the production of males by other workers is expected when egg laying by workers decreases the average inclusive fitness of individual group members. This may result (i) from the relatedness of workers being lower to worker than to queen derived males or (ii) from a lowered overall colony efficiency. Whereas good evidence exists for policing behavior caused by genetic conflicts, policing caused by efficiency factors has not been demonstrated. We investigated the regulation of reproduction in the ant Platythyrea punctata, a species in which colonies are clones because workers are capable of producing female offspring by thelytokous parthenogenesis. Reproductive conflicts resulting from differences in genetic relatedness are therefore not expected, but uncontrolled reproduction by all workers could lead to the destruction of sociality. Here we show that worker policing by aggressive attacks against additionally reproducing workers keeps the number of reproducing workers low. Furthermore, through experimental manipulation of the number of brood items per colony, we show that worker policing can enhance group efficiency. PMID- 14557540 TI - HIV-1 in genital tract and plasma of women: compartmentalization of viral sequences, coreceptor usage, and glycosylation. AB - Worldwide, 90% of HIV-1 infections are transmitted heterosexually. Because the genital mucosa are the sites of initial contact with HIV-1 for most exposed individuals, study of the virus from the genital tract is critical for the development of vaccines and therapeutics. Previous analyses of HIV-1 in various tissues have documented compartmentalization of viral genomes. Whether compartmentalization was associated with viral phenotypic differences or immune status, however, was not well understood. We compared HIV-1 gp120 env sequences from the genital tract and plasma of 12 women. Eight women displayed compartmentalized HIV-1 RNA genomes, with viral sequences from each site that were clearly discrete, yet phylogenetically related. The remaining four exhibited env sequences that were intermingled between the two sites. Women with compartmentalized HIV-1 genomes had higher CD4+ cell counts than those displaying intermingled strains (P = 0.02). Intrapatient HIV-1 recombinants comprising sequences that were characteristic of both sites were identified. We next compared viral phenotypes in each compartment. HIV-1 coreceptor usage was often compartmentalized (P 0.01). The number of N-linked glycosylation sites, associated with neutralization resistance, also differed between compartments (P < 0.01). Furthermore, disparities between the density of gp120 glycosylations in each compartment correlated with higher CD4+ counts (P = 0.03). These data demonstrate that the genital tract and plasma can harbor populations of replicating HIV-1 with different phenotypes. The association of higher CD4+ cell counts with compartmentalization of viral genomes and density of gp120 glycosylations suggests that the immune response influences the development of viral genotypes in each compartment. These findings are relevant to the prevention and control of HIV-1 infection. PMID- 14557543 TI - Genomewide screening for fusogenic human endogenous retrovirus envelopes identifies syncytin 2, a gene conserved on primate evolution. AB - Screening human sequence databases for endogenous retroviral elements with coding envelope genes has revealed 16 candidate genes that we assayed for their fusogenic properties. All 16 genes were cloned in a eukaryotic expression vector and assayed for cell-cell fusion by using a large panel of mammalian cells in transient transfection assays. Fusion was observed for two human endogenous retrovirus (HERV) envelopes, the previously characterized HERV-W envelope, also called syncytin, and a previously uncharacterized gene from the HERV-FRD family. Cells prone to env-mediated fusion were different for the two envelopes, indicating different receptor usage. A search for the FRDenv gene in primates indicated that the corresponding proviral element is present in all simians, from New World monkeys to humans, being absent only in prosimians. Cloning of the corresponding env genes in simians disclosed conservation of the fully coding status of the gene, and most remarkably, conservation of its fusogenic property. Finally, a Northern blot analysis for the expression of the FRD family among a series of human tissues demonstrated specific expression in the placenta, as previously demonstrated for the other fusogenic human envelope of the HERV-W family. Altogether, the present data have identified a previously uncharacterized envelope (that we propose to name syncytin 2 after renaming syncytin as syncytin 1) with a potential role in placenta formation, and the identification of the complete set of retroviral envelopes with fusogenic properties now allows a definite analysis of the possible role of HERV in this physiological process, via classical genetic approaches. PMID- 14557544 TI - Brain-derived neurotrophic factor-dependent unmasking of "silent" synapses in the developing mouse barrel cortex. AB - Brain-derived neurotrophic factor (BDNF) is a critical modulator of central synaptic functions such as long-term potentiation in the hippocampal and visual cortex. Little is known, however, about its role in the development of excitatory glutamatergic synapses in vivo. We investigated the development of N-methyl-D aspartate (NMDA) receptor (NMDAR)-only synapses (silent synapses) and found that silent synapses were prominent in acute thalamocortical brain slices from BDNF knockout mice even after the critical period. These synapses could be partially converted to alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR)-containing ones by adding back BDNF alone to the slice or fully converted to together with electric stimulation without affecting NMDAR transmission. Electric stimulation alone was ineffective under the BDNF knockout background. Postsynaptically applied TrkB kinase inhibitor or calcium-chelating reagent blocked this conversion. Furthermore, the AMPAR C-terminal peptides essential for interaction with PDZ proteins postsynaptically prevented the unmasking of silent synapses. These results suggest that endogenous BDNF and neuronal activity synergistically activate AMPAR trafficking into synaptic sites. PMID- 14557545 TI - Combinatorial discovery of two-photon photoremovable protecting groups. AB - A design principle for a two-photon photochemically removable protecting group based on sequential one-photon processes has been established. The expected performance of such groups in spatially directed photoactivation/photodeprotection has been shown by a kinetic analysis. One particular molecular class fitting into this design, the nitrobenzyl ethers of o hydroxycinnamates, has been presented. An initial demonstration of two-photon deprotection of one such group prompted further optimization with respect to photochemical deprotection rate. This was accomplished by the preparation and screening of a 135-member indexed combinatorial library. Optimum performance for lambda >350 nm deprotection in organic solvent was found with 4,5-dialkoxy and cyano substitution in the nitrobenzyl group and 4-methoxy substitution in the cinnamate. PMID- 14557546 TI - Contrasting patterns of expression of transcription factors in pancreatic alpha and beta cells. AB - Pancreatic alpha and beta cells are derived from the same progenitors but play opposing roles in the control of glucose homeostasis. Disturbances in their function are associated with diabetes mellitus. To identify many of the proteins that define their unique pathways of differentiation and functional features, we have analyzed patterns of gene expression in alphaTC1.6 vs. MIN6 cell lines by using oligonucleotide microarrays. Approximately 9-10% of >11,000 transcripts examined showed significant differences between the two cell types. Of >700 known transcripts enriched in either cell type, transcription factors and their regulators (TFR) was one of the most significantly different categories. Ninety six members of the basic zipper, basic helix-loop-helix, homeodomain, zinc finger, high mobility group, and other transcription factor families were enriched in alpha cells; in contrast, homeodomain proteins accounted for 51% of a total of 45 TFRs enriched in beta cells. Our analysis thus highlights fundamental differences in expression of TFR subtypes within these functionally distinct islet cell types. Interestingly, the alpha cells appear to express a large proportion of factors associated with progenitor or stem-type cells, perhaps reflecting their earlier appearance during pancreatic development. The implications of these findings for a better understanding of alpha and beta cell dysfunction in diabetes mellitus are also considered. PMID- 14557547 TI - The primary mechanism of attenuation of bacillus Calmette-Guerin is a loss of secreted lytic function required for invasion of lung interstitial tissue. AB - Tuberculosis remains a leading cause of death worldwide, despite the availability of effective chemotherapy and a vaccine. Bacillus Calmette-Guerin (BCG), the tuberculosis vaccine, is an attenuated mutant of Mycobacterium bovis that was isolated after serial subcultures, yet the functional basis for this attenuation has never been elucidated. A single region (RD1), which is absent in all BCG substrains, was deleted from virulent M. bovis and Mycobacterium tuberculosis strains, and the resulting DeltaRD1 mutants were significantly attenuated for virulence in both immunocompromised and immunocompetent mice. The M. tuberculosis DeltaRD1 mutants were also shown to protect mice against aerosol challenge, in a similar manner to BCG. Interestingly, the DeltaRD1 mutants failed to cause cytolysis of pneumocytes, a phenotype that had been previously used to distinguish virulent M. tuberculosis from BCG. A specific transposon mutation, which disrupts the Rv3874 Rv3875 (cfp-10 esat-6) operon of RD1, also caused loss of the cytolytic phenotype in both pneumocytes and macrophages. This mutation resulted in the attenuation of virulence in mice, as the result of reduced tissue invasiveness. Moreover, specific deletion of each transcriptional unit of RD1 revealed that three independent transcriptional units are required for virulence, two of which are involved in the secretion of ESAT-6 (6-kDa early secretory antigenic target). We conclude that the primary attenuating mechanism of bacillus Calmette-Guerin is the loss of cytolytic activity mediated by secreted ESAT-6, which results in reduced tissue invasiveness. PMID- 14557548 TI - MADS box genes control vernalization-induced flowering in cereals. AB - By comparing expression levels of MADS box transcription factor genes between near-isogenic winter and spring lines of bread wheat, Triticum aestivum, we have identified WAP1 as the probable candidate for the Vrn-1 gene, the major locus controlling the vernalization flowering response in wheat. WAP1 is strongly expressed in spring wheats and moderately expressed in semispring wheats, but is not expressed in winter wheat plants that have not been exposed to vernalization treatment. Vernalization promotes flowering in winter wheats and strongly induces expression of WAP1. WAP1 is located on chromosome 5 in wheat and, by synteny with other cereal genomes, is likely to be collocated with Vrn-1. These results in hexaploid bread wheat cultivars extend the conclusion made by Yan et al. [Yan, L., Loukoianov, A., Tranquilli, G., Helguera, M., Fahima, T. & Dubcovsky, J. (2003) Proc. Natl. Acad. Sci. USA 100, 6263-6268] in the diploid wheat progenitor Triticum monococcum that WAP1 (TmAP1) corresponds to the Vrn-1 gene. The barley homologue of WAP1, BM5, shows a similar pattern of expression to WAP1 and TmAP1. BM5 is not expressed in winter barleys that have not been vernalized, but as with WAP1, expression of BM5 is strongly induced by vernalization treatment. In spring barleys, the level of BM5 expression is determined by interactions between the Vrn-H1 locus and a second locus for spring habit, Vrn-H2. There is now evidence that AP1-like genes determine the time of flowering in a range of cereal and grass species. PMID- 14557549 TI - DNA-repair protein hHR23a alters its protein structure upon binding proteasomal subunit S5a. AB - The Rad23 family of proteins, including the human homologs hHR23a and hHR23b, stimulates nucleotide excision repair and has been shown to provide a novel link between proteasome-mediated protein degradation and DNA repair. In this work, we illustrate how the proteasomal subunit S5a regulates hHR23a protein structure. By using NMR spectroscopy, we have elucidated the structure and dynamic properties of the 40-kDa hHR23a protein and show it to contain four structured domains connected by flexible linker regions. In addition, we reveal that these domains interact in an intramolecular fashion, and by using residual dipolar coupling data in combination with chemical shift perturbation analysis, we present the hHR23a structure. By itself, hHR23a adopts a closed conformation defined by the interaction of an N-terminal ubiquitin-like domain with two ubiquitin-associated domains. Interestingly, binding of the proteasomal subunit S5a disrupts the hHR23a interdomain interactions and thereby causes it to adopt an opened conformation. PMID- 14557551 TI - Mitochondrial disorders come full circle. PMID- 14557550 TI - Differential regulation of midbrain dopaminergic neuron development by Wnt-1, Wnt 3a, and Wnt-5a. AB - The Wnts are a family of glycoproteins that regulate cell proliferation, fate decisions, and differentiation. In our study, we examined the contribution of Wnts to the development of ventral midbrain (VM) dopaminergic (DA) neurons. Our results show that beta-catenin is expressed in DA precursor cells and that beta catenin signaling takes place in these cells, as assessed in TOPGAL [Tcf optimal promoter beta-galactosidase] reporter mice. We also found that Wnt-1, -3a, and 5a expression is differentially regulated during development and that partially purified Wnts distinctively regulate VM development. Wnt-3a promoted the proliferation of precursor cells expressing the orphan nuclear receptor-related factor 1 (Nurr1) but did not increase the number of tyrosine hydroxylase-positive neurons. Instead, Wnt-1 and -5a increased the number of rat midbrain DA neurons in rat embryonic day 14.5 precursor cultures by two distinct mechanisms. Wnt-1 predominantly increased the proliferation of Nurr1+ precursors, up-regulated cyclins D1 and D3, and down-regulated p27 and p57 mRNAs. In contrast, Wnt-5a primarily increased the proportion of Nurr1+ precursors that acquired a neuronal DA phenotype and up-regulated the expression of Ptx3 and c-ret mRNA. Moreover, the soluble cysteine-rich domain of Frizzled-8 (a Wnt inhibitor) blocked endogenous Wnts and the effects of Wnt-1 and -5a on proliferation and the acquisition of a DA phenotype in precursor cultures. These findings indicate that Wnts are key regulators of proliferation and differentiation of DA precursors during VM neurogenesis and that different Wnts have specific and unique activity profiles. PMID- 14557552 TI - TMS: using brain plasticity to treat chronic poststroke symptoms. PMID- 14557554 TI - Hemispherectomy for intractable unihemispheric epilepsy etiology vs outcome. AB - BACKGROUND: Surgical removal of one hemisphere has been performed for several decades to treat intractable unihemispheric epilepsy. A prior case series focused on the outcomes after 58 surgeries at Johns Hopkins Hospital in 1997. This series, and an additional 53 cases, were reviewed to bring the outcomes up to date. METHODS: Charts of the 111 patients undergoing hemidecortications at the Pediatric Epilepsy Center from 1975 to 2001 were reviewed and families were contacted. Three children died in the immediate perioperative period and three were lost to follow-up immediately after surgery. Follow-up ranged from 3 months to 22 years. RESULTS: Two children died several years later due to intractable seizures. Overall, 65% are seizure-free, 21% have occasional, non-handicapping seizures, and 14% have troublesome seizures. Eighty percent are on one anticonvulsant or none and 89% are able to walk without assistance. Etiology strongly predicted seizure outcome. Patients with migrational disorders are less likely to be seizure-free than all other etiologies (predominantly Rasmussen and congenital vascular injuries) combined (51% vs 71%, p = 0.05). CONCLUSIONS: Hemidecortication continues to be a beneficial procedure in reducing seizure frequency in cases of unilateral cortical epilepsy. Fewer children with migrational disorders are seizure-free. PMID- 14557555 TI - Autoantibodies to NMDA receptor in patients with chronic forms of epilepsia partialis continua. AB - BACKGROUND: Antibody-mediated and cytotoxic T cell-mediated pathogenicity have been implicated as the autoimmune pathophysiologic mechanisms in Rasmussen's encephalitis. METHODS: The authors investigated autoantibodies against the NMDA glutamate receptor (GluR) epsilon2 subunit and their epitopes in serum and CSF samples from 15 patients with chronic epilepsia partialis continua (EPC), 17 with West syndrome, 10 with Lennox-Gastaut syndrome, and 11 control subjects. RESULTS: In 15 patients with chronic EPC, we detected NMDA-type GluR epsilon2 autoantibodies in histologically proven Rasmussen's encephalitis (3/3 patients), clinical Rasmussen's encephalitis (6/7 patients), acute encephalitis/encephalopathy (2/3 patients), and nonprogressive EPC (2/2 patients). Serum IgM autoantibodies were found in the early phase of EPC and became negative later in four patients. The autoantibodies were not detected in West syndrome, Lennox-Gastaut syndrome, or controls. Among 10 patients with histologically proven or clinical Rasmussen's encephalitis, epitope analyses showed that the autoantibodies were predominantly against C-terminal epitopes and rarely against N-terminal epitope, with inconsistency in profile during the courses of disease. Epitope recognition spectrum of autoantibodies was broader in CSF than in serum, and the serum or CSF profile showed an increase in number of epitopes as disease progressed in some patients. CONCLUSIONS: The presence of autoantibodies against NMDA GluR epsilon2 suggests autoimmune pathologic mechanisms but is not a hallmark of Rasmussen's encephalitis. Patients with Rasmussen's encephalitis may have autoantibodies against several neural molecules, and these autoantibodies may be produced in the CNS after cytotoxic T cell-mediated neuronal damage. PMID- 14557556 TI - Proton MRS reveals frontal lobe metabolite abnormalities in idiopathic generalized epilepsy. AB - OBJECTIVE: To assess gamma-aminobutyric acid (GABA) plus homocarnosine (GABA+) and glutamate plus glutamine (GLX) concentrations in the frontal lobes of patients with idiopathic generalized epilepsy (IGE). METHODS: Twenty-one patients and 17 healthy volunteers were studied. A single voxel was prescribed in each frontal lobe for each subject. Point-resolved spectroscopy (PRESS)-localized short echo time MR spectroscopy (MRS) was performed to measure GLX and the metabolites N-acetylaspartate plus N-acetylaspartylglutamate (NAAt), creatine and phosphocreatine (Cr), choline-containing compounds (Cho), and myo-inositol (Ins). A double quantum GABA filter was used to measure GABA+. Segmented T1-weighted images gave the tissue composition of the prescribed voxel. RESULTS: Group comparisons showed elevation of GLX and reduction of NAAt in the patient group (p < 0.05). The metabolite ratios GLX/NAAt and GLX/Ins also showed elevation in IGE (p = 0.01). No group effect was observed for GABA+, Cr, or Cho. Ins concentrations were not significantly reduced in the patient group but were less in the subgroup of patients who were taking sodium valproate. CONCLUSIONS: IGE was associated with bilateral frontal lobe metabolite changes. Elevation in GLX was observed, which may imply increased neuronal excitability, whereas reduction in NAAt suggests reduced overall neuronal numbers or neuronal dysfunction. PMID- 14557558 TI - Possible phenotypic dosage effect in patients compound heterozygous for FSHD sized 4q35 alleles. AB - OBJECTIVE: Autosomal dominant facioscapulohumeral muscular dystrophy (FSHD) is associated with a contraction of the D4Z4 repeat array on chromosome 4. So far, homozygosity or compound heterozygosity for FSHD alleles has not been described, and it has been debated whether the absence of such subjects is because of the rarity or the lethality of the disorder. METHODS: Two unrelated families in which the probands are compound heterozygous for two FSHD-sized alleles were studied. Clinical examination, pulsed-field gel electrophoresis (PFGE) studies of DNA with probes proximal and distal to D4Z4, and cytogenetic analysis of metaphase chromosomes by FISH were performed. RESULTS: Complementary molecular and cytogenetic approaches confirmed the chromosome 4qA origin of all FSHD-sized repeat arrays that segregate in the families. CONCLUSIONS: Heterozygosity for FSHD-sized alleles is compatible with life in men and women. A possible dosage effect was observed in both probands in whom each 4qA allele contributed to the FSHD phenotype. Because at least one of the FSHD alleles in both families showed an unusual low penetrance, the authors propose that susceptibility for FSHD is partly determined by intrinsic properties of the disease allele other than the residual D4Z4 repeat size alone. PMID- 14557557 TI - Remarkable infidelity of polymerase gammaA associated with mutations in POLG1 exonuclease domain. AB - OBJECTIVE: To better understand the still unknown pathologic mechanism involved in the accumulation of multiple mtDNA deletions in stable tissues. METHODS: A large-scale screening of mtDNA molecules from skeletal muscle was performed in 14 patients with progressive external ophthalmoplegia (PEO) and 2 patients with mitochondrial neurogastrointestinal encephalomyopathy carrying mutations on ANT1, C10ORF2 or POLG1, and TP genes. RESULTS: Patients with at least one mutation in the exonuclease domain of POLG1 showed the highest frequency of individually rare point mutations only in the mtDNA control region; in addition, high levels, in terms of frequency and heteroplasmy, of recurrent mutations (A189G, T408A, and T414G) and alterations affecting the (HT)D310 region were detectable in many of the patients. Two homozygous POLG1 mutations, within the exonuclease domain, were able to induce an increased mutational burden also in fibroblasts from patients with PEO. CONCLUSIONS: Specific POLG1 mutations directly affect the integrity of the mtDNA by reducing its proof-reading exonuclease activity, resulting in the accumulation of heteroplasmic levels of both randomly rare and recurrent point mutations in the skeletal muscle tissue and fibroblasts. PMID- 14557559 TI - Cold induces shifts of voltage dependence in mutant SCN4A, causing hypokalemic periodic paralysis. AB - BACKGROUND: The authors reported a mutation, P1158S, of the human skeletal muscle sodium channel gene (SCN4A) in a family with cold-induced hypokalemic periodic paralysis (hypoKPP) and myotonia. OBJECTIVE: To identify mechanisms of temperature dependency in this channelopathy. METHODS: Using the amphotericin B perforated patch clamp method, sodium currents were recorded at 22 and 32 degrees C from the wild-type (WT) and P1158S mutant SCN4A expressed in tsA201 cells. Computer simulation was performed, incorporating the gating parameters of the P1158S mutant SCN4A. RESULTS: P1158S mutant SCN4A exhibited hyperpolarizing shifts in voltage dependence of both activation and inactivation curves at a cold temperature and a slower rate of inactivation than the WT. Computer simulation reproduced the abnormal skeletal muscle electrical activities of both paralysis at a low potassium concentration in the cold and myotonia at a normal potassium concentration. CONCLUSIONS: Both paralysis and myotonia are attributable to the biophysical properties of the SCN4A mutation associated with hypoKPP. This is the first report of an SCN4A mutation that exhibits temperature-dependent shifts of voltage dependence in sodium channel gating. PMID- 14557560 TI - Deep brain stimulation of the VIM thalamic nucleus modifies several features of essential tremor. AB - BACKGROUND: Pharmacologic interventions (e.g., beta blockers) and thalamic lesions have failed to alter the pathophysiology of essential tremor (ET) beyond a reduction in tremor amplitude. Deep brain stimulation (DBS) of the ventral intermediate (VIM) nucleus of the thalamus successfully reduces tremor rating scores. It is unknown how VIM DBS alters the pathophysiologic characteristics of ET. OBJECTIVE: To determine the effects of VIM DBS on the neurophysiologic characteristics of ET. METHODS: Hand tremor and EMG activity of forearm extensor and flexor muscles were recorded in six patients with ET ON-DBS and OFF-DBS and from six age- and sex-matched control subjects. Hand tremor was assessed across different inertial loads. The amplitude, frequency, regularity, and tremor-EMG coherence were analyzed. RESULTS: VIM DBS reduced the amplitude, increased the frequency, decreased the regularity, and reduced the 1 to 8 Hz tremor-EMG coherence of ET. ON-DBS, patients with ET had greater tremor amplitude, lower frequency, more regularity, and greater tremor-EMG coherence compared to control subjects. CONCLUSIONS: Whereas pharmacologic and thalamic lesions have previously failed to change characteristics of ET beyond amplitude reduction, VIM DBS modified multiple features of ET. The changes in ET after VIM DBS provide strong evidence for clinical efficacy. PMID- 14557561 TI - Creatine supplementation in Huntington's disease: a placebo-controlled pilot trial. AB - OBJECTIVE: To evaluate the effect of creatine (Cr) supplementation (5 g/day) in Huntington's disease (HD). METHODS: A 1-year double-blind placebo-controlled study was performed in 41 patients with HD (stage I through III). At baseline and after 6 and 12 months, the functional, neuromuscular, and cognitive status of the patients was assessed by a test battery that consisted of 1) the Unified Huntington's Disease Rating Scale (UHDRS), 2) an exercise test on an isokinetic dynamometer to assess strength of the elbow flexor muscles, 3) a maximal exercise test on a bicycle ergometer to evaluate cardiorespiratory fitness, and 4) a test to assess bimanual coordination ability. Following the baseline measurements, the subjects were assigned to either a creatine (n = 26) or a placebo group (n = 15). RESULTS: Scores on the functional checklist of the UHDRS (p < 0.05), maximal static torque (p < 0.05), and peak oxygen uptake (p < 0.05) decreased from the start to the end of the study, independent of the treatment received. Cognitive functioning, bimanual coordination ability, and general motor function (total motor scale, UHDRS) did not change from baseline to 1 year in either group. CONCLUSION: One year of Cr intake, at a rate that can improve muscle functional capacity in healthy subjects and patients with neuromuscular disease (5 g/day), did not improve functional, neuromuscular, and cognitive status in patients with stage I to III HD. PMID- 14557562 TI - Risk of action tremor in relatives of tremor-dominant and postural instability gait disorder PD. AB - BACKGROUND: Action tremor may be more prevalent in relatives of patients with Parkinson's disease (PD) than in relatives of control subjects. This tremor could represent mild PD or essential tremor. An estimate of the risk of this condition in families of patients with PD is important when studying the genetics of PD. OBJECTIVE: S: To determine the risk of action tremor in first-degree relatives of probands with tremor-dominant PD (TD-PD) and postural instability gait disorder PD (PIGD-PD) compared with first-degree relatives of control probands. METHODS: PD and control probands participated in a familial aggregation study of PD. The presence of action tremor in their relatives was ascertained from reports of one or more informants. Relatives who met diagnostic criteria for PD were excluded. Cox proportional hazards models adjusting for gender, education, race, and vital status (dead vs alive) of the relatives were used to assess the relative risk (RR) of action tremor in first-degree relatives of PD probands vs first-degree relatives of control probands. RESULTS: There were 487 PD probands, 409 control probands, and 5,563 relatives. The risk of action tremor was higher in the relatives of TD-PD probands than in the relatives of control probands (RR = 2.14; 95% CI = 1.53 to 2.98) but not in the relatives of PIGD-PD probands compared with the relatives of control probands (RR = 1.81; 95% CI = 0.66 to 5.02). CONCLUSION: The risk of action tremor was increased in the relatives of PD probands, particularly when they had TD-PD. Whether the tremor in these relatives represents essential tremor or an isolated manifestation of PD requires further investigation. PMID- 14557563 TI - Objective assessments of longitudinal outcome in Gilles de la Tourette's syndrome. AB - OBJECTIVE: To define the long-term outcome in Gilles de la Tourette syndrome (GTS) using objective rating measures. BACKGROUND: Previous historical studies suggest spontaneous improvement of tic symptoms after adolescence, but objective longitudinal data are limited. METHODS: The authors reviewed all videotapes in their database (1978 through 1991) of children with GTS (ages 8 to 14) who were seen in their tertiary care movement disorder center and underwent a standardized 5-minute filming protocol (n = 56). Through multiple contact methods, they successfully located 36 of these patients, who are now adults (age >20 years), and recruited 31 (28 men and 3 women) to volunteer for a second videotape and in person assessment. A blinded rater evaluated the 62 tapes and rated five tic domains: body areas involved, motor and phonic tic frequency, and motor and phonic tic severity. Using standardized GTS videotape rating scale and Wilcoxon signed-rank tests with Bonferroni correction for multiple comparisons, the authors compared the two videotapes for each tic domain as well as the composite tic disability score. RESULTS: Ninety percent of adult patients still had tics. Adult patients who considered themselves tic-free were often inaccurate in their self-assessment: 50% had objective evidence of tics. Mean objective tic disability diminished in comparison to childhood (mean composite tic disability score childhood 9.58 vs adulthood 7.52, p = 0.014). All domains improved by adulthood, and significant improvements occurred in motor tic severity (p = 0.008). The improvements in tic disability did not relate to medication use, as only 13% of adults received medications for tics, compared with 81% of children. CONCLUSIONS: In GTS syndrome, tics objectively improve over time but most adults have persistent tics. PMID- 14557564 TI - A randomized, double-blind, placebo controlled study on analgesic effects of botulinum toxin A. AB - OBJECTIVE: Botulinum toxin type A (BTXA) is used to treat neurologic disorders associated with increased muscle tone. Its use is often associated with pain relief. METHODS: A possible direct analgesic effect of BTXA on C and Adelta fibers was studied on 16 healthy volunteers receiving 30 U BTXA into one forearm and pure saline into the other. To exclude the secondary effect due to muscular tone reduction, BTXA was injected intradermally. Thermal sensory testing of heat pain (threshold and tolerance) and neuroselective current sensory testing of current pain threshold/tolerance were performed at baseline and 3, 14, and 28 days after treatment. Thereafter, on day 28, capsaicin was administered simultaneously into both forearms to evaluate a possible peripheral effect and central effect on pain processing and on the axon reflex flare. RESULTS: The authors observed no significant difference in any of the perception outcome measures between BTXA and placebo pretreated areas. Flare areas as a result of the release of neuropeptides after capsaicin application showed no differences. CONCLUSIONS: The results suggest that pain reduction after BTXA treatment is mediated through its effect on muscle tone rather than a direct analgesic effect. PMID- 14557565 TI - The impact of parkinsonism on costs of care in patients with AD and dementia with Lewy bodies. AB - OBJECTIVE: To compare the annual direct costs for patients with Alzheimer's disease (AD) and no parkinsonism (AD), patients with AD and parkinsonism (AD/P), and patients with dementia with Lewy bodies (DLB). For the entire sample, the authors examined the incremental costs associated with increasing parkinsonism. METHODS: Cross-sectional comparisons of 1 year of direct costs were done. Fifteen patients met criteria for DLB, and 133 met criteria for probable AD, of whom 39 had signs of parkinsonism and 94 did not. Patients' caregivers reported on the patient's use of health care services, receipt of unpaid care, and comorbid medical conditions. Severity of cognitive impairment and parkinsonism were determined during patient examinations. Costs were estimated by multiplying utilization data by a unit cost for each type of care. Costs were compared after adjustment for covariates using multiple regression equations. RESULTS: After adjusting for important covariates, patients with AD/P or DLB had significantly higher annual direct costs than patients with AD. The average adjusted increases in costs above the AD baseline costs were 7,119 dollars (AD/P) and 13,754 dollars (DLB) for formal direct costs and 7,394 dollars (AD/P) and 19,564 dollars (DLB) for total direct costs. Models for the entire sample estimated that a 1-point increase in a parkinsonism scale would result in an annual increase of 784 dollars in formal costs and 827 dollars in total costs of care. CONCLUSIONS: Patients with Alzheimer's disease and parkinsonism or dementia with Lewy bodies have significantly higher formal and total direct costs of care than patients with Alzheimer's disease. Signs of parkinsonism in patients with degenerative dementias are significant independent predictors of costs of care. PMID- 14557566 TI - Changes in DWI and MRS associated with white matter hyperintensities in elderly subjects. AB - OBJECTIVE: To assess normal-appearing white matter (NAWM) characteristics by magnetic resonance spectroscopy (MRS) and diffusion-weighted imaging (DWI) in elderly subjects. METHODS: The authors studied 60 volunteers (mean age 72.6 years; SD 4.7; range 64 to 84 years) without signs of neurologic illness. They used DWI and spectroscopic imaging to investigate whether there were changes in the NAWM that related to the presence of white matter hyperintensities (WMH). RESULTS: The authors found a correlation (p < 0.001) between the apparent diffusion coefficient in the NAWM and the total volume of WMH. The metabolite ratios N-acetylaspartate/creatine and N-acetylaspartate/choline of the NAWM also correlated significantly with total WMH volume. These correlations were independent of age. CONCLUSIONS: Damage associated with WMH is detectable in NAWM. PMID- 14557567 TI - Neuromotor abnormalities and risk for psychosis in Alzheimer's disease. AB - BACKGROUND: Cross-sectional studies in Alzheimer's disease (AD) show a strong relationship between extrapyramidal motor signs and presence of psychosis, yet it remains unclear whether neuromotor abnormalities precede and therefore can predict development of psychosis in AD. OBJECTIVE: To identify cognitive and motor risk factors for the development of psychosis in patients with AD. METHODS: Baseline clinical motor ratings and instrumental measures of neuromuscular function were obtained from 54 nonpsychotic patients with AD who were evaluated annually for 2 years for the development of psychosis. Survival analyses were performed to identify incidence and risks associated with psychosis. RESULTS: The incidence of new onset psychosis in our sample was 32.5% in 2 years. Patients with abnormal agonist muscle burst amplitudes during rapid alternating movements of the hand were more likely to develop psychosis than those without (OR = 4.31; p = 0.007). Women with AD also had a higher risk of developing psychosis within 2 years than men (OR = 1.33; p = 0.01). CONCLUSIONS: Using simple noninvasive instrumental procedures for assessing neuromotor function, it may be possible to identify an individual's level of risk for developing psychosis during the course of AD. PMID- 14557568 TI - White matter disease and sleep-disordered breathing after acute stroke. AB - OBJECTIVE: To assess sleep-disordered breathing (SDB) in patients with acute stroke and its relationship to prestroke cerebrovascular disease, particularly leukoariosis. METHODS: The authors studied SDB and CT evidence of prestroke cerebrovascular disease, nonacute brain infarction, and white matter disease (WMD) in 78 previously independent patients with acute stroke. Subjects underwent respiratory sleep studies to determine the Apnea-Hypopnea Index (AHI) and Desaturation Index (DI) 7 to 14 days following stroke. CT scans were evaluated for severity of acute prestroke cerebrovascular disease and WMD severity. RESULTS: Prestroke cerebrovascular disease was present in 49 (63%) patients and was associated with worse SDB (mean AHI 35 vs 23, p < 0.01; mean DI 28 vs 18, p < 0.05), particularly in men (mean AHI 43 vs 27, p < 0.01). WMD severity correlated with AHI (r = 0.26, p < 0.05) and age (r = 0.56, p < 0.01). WMD severity in frontal and basal ganglia areas showed the strongest association. On multivariate analysis, WMD severity correlated independently with AHI (R(2) = 0.07, p < 0.05). CONCLUSIONS: The presence of prestroke cerebrovascular disease and severity of WMD are associated with worse SDB. These findings suggest that either white matter is particularly vulnerable to the hypoxia and blood pressure variability associated with SDB or that WMD is a major factor exacerbating SDB following stroke. PMID- 14557569 TI - Stroke risk in anticoagulated patients with atrial fibrillation undergoing endoscopy. AB - OBJECTIVE: To determine the risk of stroke in anticoagulated patients with atrial fibrillation (AF) when anticoagulation is adjusted for bronchoscopy, gastroscopy, or colonoscopy, and to identify factors that could modify this risk. METHODS: The authors reviewed patients with AF undergoing endoscopies between 1995 and 2001, with specific analysis of patients with AF in whom anticoagulation was adjusted for the procedures. The authors calculated the stroke rate within 30 days of the procedures. RESULTS: Twelve strokes occurred in 987 patients undergoing 1,137 procedures (1.06%/procedure). The risk ranged from 0.31% for patients with nonvalvular AF undergoing routine procedures to 2.93% for complex patients undergoing endoscopies combined with other procedures or with comorbid illnesses. Patients with stroke were more likely to be complex (7/12 vs 219/975, p = 0.04); to be older than 80 years (6/12 vs 187/975, p = 0.017); to have a history of stroke (7/12 vs 194/975, p = 0.004), hypertension (10/12 vs 508/975, p = 0.04), or hyperlipidemia (9/12 vs 334/975, p = 0.005); or to have a family history of vascular disease (10/12 vs 502/975, p = 0.039). CONCLUSIONS: The risk of stroke in patients with AF whose anticoagulation is adjusted for endoscopies is low, but almost tenfold higher in patients with complex clinical situations. Age, history of stroke, hypertension, hyperlipidemia, and family history of vascular disease may increase the risk of stroke. PMID- 14557570 TI - The "double panda sign" in Wilson's disease. PMID- 14557571 TI - The neurologist and Harvard criteria for brain death. AB - The Harvard ad hoc committee to examine the definition of brain death, chaired by anesthesiologist Beecher, published their report in 1968. Throughout the years, the committee has been a target of criticism. However, the workings of the ad hoc committee have not been carefully evaluated, and could provide insight into the role of the neurologists and their interaction with contemporaries. Review of the original files and interviews with three surviving members-Adams, Potter, and Murray-revealed 1) a major role and original contributions of the neurologists Schwab and Adams in defining criteria, and 2) a swift and decisive working committee that formulated a document aimed at simplicity and accuracy, and without special interest. PMID- 14557572 TI - Plasticity of motor cortex excitability induced by rehabilitation therapy for writing. AB - The mechanisms of rehabilitation-induced plasticity in the motor system after stroke are not defined. The authors studied seven patients with residual poststroke agraphia, aphasia, and right hemiparesis. After a 40-minute rehabilitation therapy that promoted use of the paretic hand for writing, the authors observed a task-specific increase in recruitment of ipsilateral corticospinal pathways. Rehabilitation aimed to increase the use of the paretic hand may induce recruitment of previously silent ipsilateral corticospinal pathways even in poorly recovered poststroke patients. PMID- 14557573 TI - Orthostatic headaches without CSF leak in postural tachycardia syndrome. AB - Four women age 17 to 28 years presented with orthostatic headaches as the most prominent feature of their symptom complex. None had CSF leak or intracranial hypotension. Autonomic studies showed evidence of orthostatic intolerance with tachycardia in all cases. Treatment of orthostatic intolerance, mainly with volume expansion, was only partially effective. Orthostatic headaches are not always caused by CSF leak or supine intracranial hypotension. Occasionally they may be the major clinical manifestation of postural tachycardia syndrome or orthostatic intolerance. PMID- 14557575 TI - Painful vascular compression syndrome of the sciatic nerve caused by gluteal varicosities. AB - The authors report three patients with chronic sciatic pain without focal neurologic deficit. Sitting or lying on the affected side provoked pain, and standing and walking relieved it. MRI revealed varicotic gluteal vessels compressing the sciatic nerve. Decompression of the nerve resulted in complete and permanent pain relief. Sciatic or buttock pain in patients with varicosities and pain provocation in the sitting or lying position suggests this neurovascular compression syndrome. PMID- 14557574 TI - Does induction of plastic change in motor cortex improve leg function after stroke? AB - Combined peripheral nerve and brain stimulation ("dual stimulation") induces changes in the excitability of normal motor cortex. The authors sought to establish whether dual stimulation would also induce motor cortex plasticity and associated functional improvements in nine stroke patients with chronic stable hemiparesis. Following 4 weeks of daily dual stimulation, improvements were seen in some neurophysiological and functional measures. This technique may offer therapeutic opportunities in some stroke patients. PMID- 14557576 TI - Charcot-Marie-Tooth disease with giant axons: a clinicopathological and genetic entity. AB - The authors report an Italian family with autosomal-dominant Charcot-Marie-Tooth disease (CMT) in which there were giant axons in the sural nerve biopsy. Linkage to the known CMT2 loci (CMT2A, CMT2B, CMT2D, CMT2F) and mutations in the known CMT2 genes (Cx32, MPZ, NEFL), GAN, NEFM, and CMT1A duplication/HNPP deletion were excluded. This family with CMT and giant axons has a pathologic and genetic entity distinct from classic CMT. PMID- 14557577 TI - SURF1 gene mutations in three cases with Leigh syndrome and cytochrome c oxidase deficiency. AB - Leigh syndrome (LS) is one of the most frequent forms of mitochondrial disease in infancy and childhood. Mutations in SURF1 have been shown to be an important cause of LS with cytochrome c oxidase (COX) deficiency. The authors have identified four pathogenic mutations including a novel, in-frame, 15-bp tandem duplication (806-820) in exon 8 and a novel 751+1G>A splice site mutation in SURF1 in three cases of LS with COX deficiency. PMID- 14557578 TI - Gammadelta T cell non-responsiveness in Campylobacter jejuni-associated Guillain Barre syndrome patients. AB - To seek evidence for a role of molecular mimicry in the induction of Guillain Barre syndrome (GBS), the authors studied Campylobacter jejuni-reactive T lymphocytes in patients with GBS. In contrast to controls, gammadelta T cells of patients with GBS with antecedent C jejuni infections failed to respond to C jejuni. Supplementing cell cultures with the cytokines interleukin-2 or interleukin-15 resulted in restoration of the gammadelta T cell proliferative response. Gammadelta T cell non-responsiveness may lead to defective regulation of antibody production, and in this way an (auto)immune response against ganglioside-like epitopes on peripheral nerve may cause GBS. PMID- 14557579 TI - Patients with severe muscle wasting are prone to develop hypoglycemia during fasting. AB - The authors investigated whether hypoglycemia develops during 23 hours of fasting in patients with Duchenne dystrophy (7 patients), spinal muscular atrophy (4 patients), and congenital myopathy (2 patients), all with residual muscle mass <10% of body weight. All patients with spinal muscular atrophy and congenital myopathy and one patient with Duchenne dystrophy, but none of six healthy subjects, developed hypoglycemia. Skeletal muscle is an important source of gluconeogenic substrates during fasting. Hypoglycemia must be considered in patients with low muscle mass, especially during surgery and febrile episodes. PMID- 14557580 TI - Assessment of a DJ-1 (PARK7) polymorphism in Finnish PD. AB - Mutations in DJ-1 are a cause of autosomal recessive parkinsonism. Polymorphism of genes implicated in hereditary forms of parkinsonism may be a predisposing factor in sporadic Parkinson's disease (PD). The authors analyzed whether a polymorphism (g.168_185del) within exon 1 of DJ-1 contributes to the risk of sporadic PD in a Finnish case-control series. This gene does not play a major role in the genetic predisposition to PD in this population. PMID- 14557581 TI - Huntington's disease--like 2 can present as chorea-acanthocytosis. AB - Three patients from a previously described family with autosomal dominant chorea acanthocytosis were found to have the CTG trinucleotide repeat expansion mutation of the junctophilin-3 gene associated with Huntington's disease-like 2 (HDL2). One of six previously identified patients with HDL2 had acanthocytosis on peripheral blood smear, suggesting that HDL2 should be considered in the differential of chorea-acanthocytosis. PMID- 14557582 TI - PS1 Alzheimer's disease family with spastic paraplegia: the search for a gene modifier. AB - PS1 mutations are associated with classic Alzheimer's disease (AD); however, some families develop AD and spastic paraplegia (SP) with brain pathology characterized by Abeta cotton wool plaques. The authors report a variant AD family with the E280Q PS1 mutation. The fact that the same PS1 mutation can be found in patients with either variant or classic AD argues in favor of the presence of a genetic modifier. The authors have excluded that this modifier effect originates from coding sequence variations in three SP genes or from a second mutation in the other AD genes. PMID- 14557583 TI - L-DOPA-induced excessive daytime sleepiness in PD: a placebo-controlled case with MSLT assessment. AB - A 74-year-old patient with idiopathic Parkinson's disease was evaluated for unintended sleep episodes that occurred after long-term treatment with 400 mg/day of L-dopa. Overnight sleep studies and multiple sleep latency testing were carried out under double-blind administration of either L-dopa or placebo. Mean sleep latency with L-dopa was 7 minutes, in contrast to a normal value of 19 minutes, 25 seconds with placebo. The authors' results suggest that L-dopa may cause daytime somnolence in some patients with Parkinson's disease. PMID- 14557584 TI - Vibration-induced median neuropathy. PMID- 14557585 TI - Seizure exacerbation associated with oxcarbazepine in idiopathic focal epilepsy of childhood. PMID- 14557586 TI - Disappearance of resting tremor after "stereotaxic" thalamic stroke. PMID- 14557587 TI - Alexander disease with serial MRS and a new mutation in the glial fibrillary acidic protein gene. PMID- 14557588 TI - Diffusion-weighted MRI abnormalities as a clue to the diagnosis of herpes simplex encephalitis. PMID- 14557589 TI - Season of birth effect in narcolepsy. PMID- 14557590 TI - Atypical Leigh syndrome associated with the D393N mutation in the mitochondrial ND5 subunit. PMID- 14557591 TI - Response of recurrent anaplastic ependymoma to a combination of tamoxifen and isotretinoin. PMID- 14557592 TI - Sensorineural hearing loss: a reversible effect of vigabatrin. PMID- 14557593 TI - Treatment of steroid-unresponsive tumefactive demyelinating disease with plasma exchange. PMID- 14557594 TI - The impact of comorbid disease and injuries on resource use and expenditures in parkinsonism. PMID- 14557595 TI - Premorbid weight, body mass, and varsity athletics in ALS. PMID- 14557596 TI - Neutralizing antibodies during treatment of secondary progressive MS with interferon beta-1b. PMID- 14557597 TI - Comparison of proteomic and genomic analyses of the human breast cancer cell line T47D and the antiestrogen-resistant derivative T47D-r. AB - In search of novel mechanisms leading to the development of antiestrogen resistance in human breast tumors, we analyzed differences in the gene and protein expression pattern of the human breast carcinoma cell line T47D and its derivative T47D-r, which is resistant toward the pure antiestrogen ZM 182780 (Faslodex trade mark, fulvestrant). Affymetrix DNA chip hybridizations on the commercially available HuGeneFL and Hu95A arrays were carried out in parallel to the proteomics analysis where the total cellular protein content of T47D or T47D r was separated on two-dimensional gels. Thirty-eight proteins were found to be reproducibly up- or down-regulated more than 2-fold in T47D-r versus T47D in the proteomics analysis. Comparison with differential mRNA analysis revealed that 19 of these were up- or down-regulated in parallel with the corresponding mRNA molecules, among which are the protease cathepsin D, the GTPases Rab11a and MxA, and the secreted protein hAG-2. For 11 proteins, the corresponding mRNA was not found to be differentially expressed, and for eight proteins an inverse regulation was found at the mRNA level. In summary, mRNA expression data, when combined with proteomic information, provide a more detailed picture of how breast cancer cells are altered in their antiestrogen-resistant compared with the antiestrogen-sensitive state. PMID- 14557598 TI - Proteomic analysis of chronic lymphocytic leukemia subtypes with mutated or unmutated Ig V(H) genes. AB - Chronic lymphocytic leukemia (CLL) is a common hematopoietic malignant disease with variable outcome. CLL has been divided into distinct groups based on whether somatic hypermutation has occurred in the variable region of the immunoglobulin heavy-chain locus or alternatively if the cells express higher levels of the CD38 protein. We have analyzed the proteome of 12 cases of CLL (six mutated (M-CLL) and six unmutated (UM-CLL) immunoglobulin heavy-chain loci; seven CD38-negative and five CD38-positive) using two-dimensional electrophoresis and mass spectrometry. Statistical evaluation using principal component analysis indicated significant differences in patterns of protein expression between the cases with and without somatic mutation. Specific proteins indicated by principal component analysis as varying between the prognostic groups were characterized using mass spectrometry. The levels of F-actin-capping protein beta subunit, 14-3-3 beta protein, and laminin-binding protein precursor were significantly increased in M CLL relative to UM-CLL. In addition, primary sequence data from tandem mass spectrometry showed that nucleophosmin was present as several protein spots in M CLL but was not detected in UM-CLL samples, suggesting that several post translationally modified forms of nucleophosmin vary between these two sample groups. No specific differences were found between CD38-positive and -negative patient samples using the same approach. The results presented show that proteomic analysis can complement other approaches in identifying proteins that may have potential value in the biological and diagnostic distinction between important clinical subtypes of CLL. PMID- 14557599 TI - Complementary analysis of the Mycobacterium tuberculosis proteome by two dimensional electrophoresis and isotope-coded affinity tag technology. AB - Classical proteomics combined two-dimensional gel electrophoresis (2-DE) for the separation and quantification of proteins in a complex mixture with mass spectrometric identification of selected proteins. More recently, the combination of liquid chromatography (LC), stable isotope tagging, and tandem mass spectrometry (MS/MS) has emerged as an alternative quantitative proteomics technology. We have analyzed the proteome of Mycobacterium tuberculosis, a major human pathogen comprising about 4,000 genes, by (i) 2-DE and mass spectrometry (MS) and by (ii) the isotope-coded affinity tag (ICAT) reagent method and MS/MS. The data obtained by either technology were compared with respect to their selectivity for certain protein types and classes and with respect to the accuracy of quantification. Initial datasets of 60,000 peptide MS/MS spectra and 1,800 spots for the ICAT-LC/MS and 2-DE/MS methods, respectively, were reduced to 280 and 108 conclusively identified and quantified proteins, respectively. ICAT LC/MS showed a clear bias for high M(r) proteins and was complemented by the 2 DE/MS method, which showed a preference for low M(r) proteins and also identified cysteine-free proteins that were transparent to the ICAT-LC/MS method. Relative quantification between two strains of the M. tuberculosis complex also revealed that the two technologies provide complementary quantitative information; whereas the ICAT-LC/MS method quantifies the sum of the protein species of one gene product, the 2-DE/MS method quantifies at the level of resolved protein species, including post-translationally modified and processed polypeptides. Our data indicate that different proteomic technologies applied to the same sample provide complementary types of information that contribute to a more complete understanding of the biological system studied. PMID- 14557601 TI - What the amygdala does and doesn't do in aversive learning. PMID- 14557602 TI - Desensitization of postsynaptic glutamate receptors contributes to high-frequency homosynaptic depression of aplysia sensorimotor connections. AB - Withdrawal reflexes of Aplysia are mediated in part by a monosynaptic circuit of sensory (SN) and motor (MN) neurons. A brief high-frequency burst of spikes in the SN produces excitatory postsynaptic potentials (EPSPs) that rapidly decrease in amplitude during the burst of activity. It is generally believed that this and other (i.e., low-frequency) forms of homosynaptic depression are entirely caused by presynaptic mechanisms (e.g., depletion of releasable transmitter). The present study examines the contribution that desensitization of postsynaptic glutamate receptors makes to homosynaptic depression. Bath application of cyclothiazide, an agent that reduces desensitization of non-NMDA glutamate receptors, reduced high-, but not low-frequency synaptic depression. Thus, a postsynaptic mechanism, desensitization of glutamate receptors, can also contribute to homosynaptic depression of sensorimotor synapses. PMID- 14557603 TI - Hemispheric dissociation of the involvement of NOS isoforms in memory for discriminated avoidance in the chick. AB - Previous research has indicated a role for both the neuronal (nNOS) and endothelial (eNOS) nitric oxide isoforms in memory formation. In addition, two distinct periods of activity of nitric oxide activity, dissociated by hemispheric localization, are implicated following passive avoidance training in the chick. In the present study, we trained black Australorp-white Leghorn chicks on a color discrimination avoidance task. Diphenyleneiodonium chloride (1 microM) or N propyl-l-arginine (50 microM) was administered into either the left or right hemisphere of the chick brain in an attempt to differentiate the effects of inhibiting eNOS or nNOS, respectively. The memory loss previously observed following administration of diphenyleneiodonium chloride between 10 and 20 min posttraining was found to be lateralized to the right hemisphere, although administration of this agent into the left hemisphere around the time of training was also amnestic. In contrast, N-propyl-l-arginine caused memory loss only when administered to the left hemisphere around the time of training. These findings suggest that activation of both eNOS and nNOS isoforms may be essential for long term memory consolidation of this task. Further, these two periods of activity are defined temporally and by hemisphere localization, although confirmation with more selective inhibitors when they become available is advised. PMID- 14557604 TI - A specific role for the human amygdala in olfactory memory. AB - The medial temporal lobe is known to play a role in the processing of olfaction and memory. The specific contribution of the human amygdala to memory for odors has not been addressed, however. The role of this region in memory for odors was assessed in patients with unilateral amygdala damage due to temporal lobectomy (n = 20; 11 left, 9 right), one patient with selective bilateral amygdala damage, and in 20 age-matched normal controls. Fifteen odors were presented, followed 1 h later by an odor-name matching test and an odor-odor recognition test. Signal detection analyses showed that both unilateral groups were impaired in their memory for matching odors with names, these patients were not significantly impaired on odor-odor recognition. Bilateral amygdala damage resulted in severe impairment in both odor-name matching as well as in odor-odor recognition memory. Importantly, none of the patients were impaired on an auditory verbal learning task, suggesting that these findings reflect a specific impairment in olfactory memory, and not merely a more general memory deficit. Taken together, the data provide neuropsychological evidence that the human amygdala is essential for olfactory memory. PMID- 14557605 TI - Emotion-induced amnesia in rats: working memory-specific impairment, corticosterone-memory correlation, and fear versus arousal effects on memory. AB - We have shown previously that psychological stress (predator exposure) impairs spatial memory in rats. We have extended that finding here to show that predator stress selectively impaired recently acquired (hippocampal-dependent) spatial working memory without affecting long-term (hippocampal-independent) spatial reference memory. We also investigated why predator exposure impairs memory. Was spatial memory impaired because of the fear-provoking aspects of predator exposure or only because the cat was a novel and arousing stimulus? If the latter possibility was correct, then any novel and arousing stimulus, independent of its emotional valence (i.e., aversive or appetitive), would impair memory. We found that spatial working memory was not impaired when the male rats were exposed to a sexually receptive female rat, a stimulus that was novel and arousing to them, but not aversive. We also found that there was an equivalent increase in serum corticosterone levels in male rats exposed to either a cat or a female rat, but only the cat-exposed rats exhibited a significant correlation between corticosterone levels and impaired memory. Overall, this series of experiments demonstrates that (1). predator stress selectively impaired working (hippocampal dependent), but not reference (hippocampal-independent), memory; (2). a fear provoking stimulus, and not merely novelty and increased arousal, impaired spatial memory; and (3). increased corticosterone levels correlated with impaired spatial working memory only under predator exposure, that is, fear-provoking conditions. PMID- 14557606 TI - Developmental changes in eyeblink conditioning and neuronal activity in the pontine nuclei. AB - Neuronal activity was recorded in the pontine nuclei of developing rats during eyeblink conditioning on postnatal days 17-18 (P17-P18) or P24-P25. A pretraining session consisted of unpaired presentations of a 300-msec tone conditioned stimulus (CS) and a 10-msec periorbital shock unconditioned stimulus (US). Five paired training sessions followed the unpaired session, consisting of 100 trials of the CS paired with the US. The rats trained on P24-P25 exhibited significantly more conditioned responses (CRs) than the rats trained on P17-P18, although both groups produced CRs by the end of training. Ontogenetic increases in pre-CS and stimulus-elicited activity in the pontine nuclei were observed during the pretraining session and after paired training. The activity of pontine units was greater on trials with CRs relative to trials without CRs in rats trained on P24 P25, but almost no CR-related modulation was observed in the pontine units of rats trained on P17-P18. The findings indicate that pontine neuronal responses to the CS and modulation of pontine activity by the cerebellum and red nucleus undergo substantial postnatal maturation. The developmental changes in pontine neuronal activity might play a significant role in the ontogeny of eyeblink conditioning. PMID- 14557607 TI - Stimulus generalization of conditioned eyelid responses produced without cerebellar cortex: implications for plasticity in the cerebellar nuclei. AB - In Pavlovian eyelid conditioning and adaptation of the vestibulo-ocular reflex, cerebellar cortex lesions fail to completely abolish previously acquired learning, indicating an additional site of plasticity in the deep cerebellar or vestibular nucleus. Three forms of plasticity are known to occur in the deep cerebellar nuclei: formation of new synapses, plasticity at existing synapses, and changes in intrinsic excitability. Only a cell-wide increase in excitability predicts that learning should generalize broadly from a training stimulus to other stimuli capable of supporting learning, whereas the alternatives predict that learning should be relatively specific to the training stimulus. Here we show that deep nucleus plasticity, as assessed by conditioned eyelid responses produced without input from the cerebellar cortex, is relatively specific to the training conditioned stimulus (CS). We trained rabbits to a tone or light CS with periorbital stimulation as the unconditioned stimulus (US), and pharmacologically disconnected the cerebellar cortex during a posttraining generalization test. The short-latency conditioned responses unmasked by this treatment showed strong decrement along the dimension of auditory frequency and did not generalize across stimulus modalities. These results cannot be explained solely by a cell-wide increase in the excitability of deep nucleus neurons, and imply that an input specific mechanism in the deep cerebellar nucleus operates as well. PMID- 14557608 TI - Role of the serotonin 5-HT(2A) receptor in learning. AB - This study reviews the role of the serotonin 5-HT2A receptor in learning as measured by the acquisition of the rabbit's classically conditioning nictitating membrane response, a component of the eyeblink response. Agonists at the 5-HT2A receptor including LSD (d-lysergic acid diethylamide) enhanced associative learning at doses that produce cognitive effects in humans. Some antagonists such as BOL (d-bromolysergic acid diethylamide), LY53,857, and ketanserin acted as neutral antagonists in that they had no effect on learning, whereas others (MDL11,939, ritanserin, and mianserin) acted as inverse agonists in that they retarded learning through an action at the 5-HT2A receptor. These results were placed in the context of what is known concerning the anatomical distribution and electrophysiological effects of 5-HT2A receptor activation in frontal cortex and hippocampus, as well as the role of cortical 5-HT2A receptors in schizophrenia. It was concluded that the 5-HT2A receptor demonstrates constitutive activity, and that variations in this activity can produce profound alterations in cognitive states. PMID- 14557609 TI - A pharmacological analysis of an associative learning task: 5-HT(1) to 5-HT(7) receptor subtypes function on a pavlovian/instrumental autoshaped memory. AB - Recent studies using both invertebrates and mammals have revealed that endogenous serotonin (5-hydroxytryptamine [5-HT]) modulates plasticity processes, including learning and memory. However, little is currently known about the mechanisms, loci, or time window of the actions of 5-HT. The aim of this review is to discuss some recent results on the effects of systemic administration of selective agonists and antagonists of 5-HT on associative learning in a Pavlovian/instrumental autoshaping (P/I-A) task in rats. The results indicate that pharmacological manipulation of 5-HT1-7 receptors or 5-HT reuptake sites might modulate memory consolidation, which is consistent with the emerging notion that 5-HT plays a key role in memory formation. PMID- 14557611 TI - Quantitation of contacts among sensory, motor, and serotonergic neurons in the pedal ganglion of aplysia. AB - Present models of long-term sensitization in Aplysia californica indicate that the enhanced behavioral response is due, at least in part, to outgrowth of sensory neurons mediating defensive withdrawal reflexes. Presumably, this outgrowth strengthens pre-existing connections by formation of new synapses with follower neurons. However, the relationship between the number of sensorimotor contacts and the physiological strength of the connection has never been examined in intact ganglia. As a first step in addressing this issue, we used confocal microscopy to examine sites of contact between sensory and motor neurons in naive animals. Our results revealed relatively few contacts between physiologically connected cells. In addition, the number of contact sites was proportional to the amplitude of the EPSP elicited in the follower motor neuron by direct stimulation of the sensory neuron. This is the first time such a correlation has been observed in the central nervous system. Serotonin is the neurotransmitter most closely examined for its role in modulating synaptic strength at the sensorimotor synapse. However, the structural relationship of serotonergic processes and sensorimotor synapses has never been examined. Surprisingly, serotonergic processes usually made contact with sensory and motor neurons at sites located relatively distant from the sensorimotor synapse. This result implies that heterosynaptic regulation is due to nondirected release of serotonin into the neuropil. PMID- 14557612 TI - Enhanced retention in the passive-avoidance task by 5-HT(1A) receptor blockade is not associated with increased activity of the central nucleus of the amygdala. AB - The effect of blockade of 5-HT1A receptors was investigated on (1). retention in a mildly aversive passive-avoidance task, and (2). spontaneous single-unit activity of central nucleus of the amygdala (CeA) neurons, a brain site implicated in modulation of retention. Systemic administration of the selective 5 HT1A antagonist NAN-190 immediately after training markedly-and dose-dependently facilitated retention in the passive-avoidance task; enhanced retention was time dependent and was not attributable to variations in wattages of shock received by animals. Systemic administration of NAN-190 had mixed effects on spontaneous single-unit activity of CeA neurons recorded extracellularly in vivo; microiontophoretic application of 5-HT, in contrast, consistently and potently suppressed CeA activity. The present findings-that 5-HT1A receptor blockade by NAN-190 (1). enhances retention in the passive-avoidance task, and (2). does not consistently increase spontaneous neuronal activity of the CeA-provide evidence that a serotonergic system tonically inhibits modulation of retention in the passive-avoidance task through activation of the 5-HT1A receptor subtype at brain sites located outside the CeA. PMID- 14557613 TI - Delay-dependent working memory impairment in young-adult and aged 5-HT1BKO mice as assessed in a radial-arm water maze. AB - Serotonin (5-HT) plays a modulatory role in mnemonic functions, especially by interacting with the cholinergic system. The 5-HT1B receptor is a key target of this interaction. The 5-HT1B receptor knockout mice were found previously to exhibit a facilitation in hippocampal-dependent spatial reference memory learning. In the present study, we submitted mice to a delayed spatial working memory task, allowing the introduction of various delays between an exposure trial and a test trial. The 5-HT1BKO and wild-type mice learned the task in a radial-arm water maze (returning to the most recent presented arm containing the escape platform), and exhibited a high level of performance at delays of 0 and 5 min. However, at the delay of 60 min, only 5-HT1BKO mice exhibited an impairment. At a delay of 90 min, all mice were impaired. Treatment by scopolamine (0.8 mg/kg) induced the same pattern of performance in wild type as did the mutation for short (5 min, no impairment) and long (60 min, impairment) delays. The 22 month-old wild-type and knockout mice exhibited an impairment at short delays (5 and 15 min). The effect of the mutation affected both young-adult and aged mice at delays of 15, 30, and 60 min. Neurobiological data show that stimulation of the 5-HT1B receptor inhibits the release of acetylcholine in the hippocampus, but stimulates this in the frontal cortex. This dual function might, at least in part, explain the opposite effect of the mutation on reference memory (facilitation) and delay-dependent working memory (impairment). These results support the idea that cholinergic-serotonergic interactions play an important role in memory processes. PMID- 14557610 TI - Multiple serotonergic mechanisms contributing to sensitization in aplysia: evidence of diverse serotonin receptor subtypes. AB - The neurotransmitter serotonin (5-HT) plays an important role in memory encoding in Aplysia. Early evidence showed that during sensitization, 5-HT activates a cyclic AMP-protein kinase A (cAMP-PKA)-dependent pathway within specific sensory neurons (SNs), which increases their excitability and facilitates synaptic transmission onto their follower motor neurons (MNs). However, recent data suggest that serotonergic modulation during sensitization is more complex and diverse. The neuronal circuits mediating defensive reflexes contain a number of interneurons that respond to 5-HT in ways opposite to those of the SNs, showing a decrease in excitability and/or synaptic depression. Moreover, in addition to acting through a cAMP-PKA pathway within SNs, 5-HT is also capable of activating a variety of other protein kinases such as protein kinase C, extracellular signal regulated kinases, and tyrosine kinases. This diversity of 5-HT responses during sensitization suggests the presence of multiple 5-HT receptor subtypes within the Aplysia central nervous system. Four 5-HT receptors have been cloned and characterized to date. Although several others probably remain to be characterized in molecular terms, especially the Gs-coupled 5-HT receptor capable of activating cAMP-PKA pathways, the multiplicity of serotonergic mechanisms recruited into action during learning in Aplysia can now be addressed from a molecular point of view. PMID- 14557615 TI - Acquisition, retention, and recall of memory after injection of RS67333, a 5 HT(4) receptor agonist, into the nucleus basalis magnocellularis of the rat. AB - The serotonin 5-HT4 subtype receptor is predominantly localized into anatomical structures linked to memory and cognition. A few experimental studies report that the acute systemic administration of selective 5-HT4 agonists has ameliorative effects on memory performance, and that these effects are reversed by contemporary administration of 5-HT4 receptor antagonists. To verify whether this procognitive action occurs via the activation of the cholinergic nucleus basalis magnocellularis (NBM)-cortical pathways, we examined the effects of RS67333, a selective partial agonist of the 5-HT4 receptor, on rat performance in a place recognition task upon local administration of the drug into the NBM area. The intra-NBM administration of RS67333 enhances the acquisition (200-500 ng/0.5 microL) and the consolidation (40-200 ng/0.5 microL) of the place recognition memory. These effects are reversed by pretreatment with the selective 5-HT4 receptor antagonist RS39604 (300 ng/0.5 microL). Conversely, the recall of memory is not affected by the 5-HT4 agonist. Our results indicate that 5-HT4 receptors located within the NBM may play a role in spatial memory and that the procognitive effect of RS67333 is due, at least in part, to the potentiation of the activity of cholinergic NBM-cortical pathways. PMID- 14557614 TI - Improvements in hippocampal-dependent learning and decremental attention in 5 HT(3) receptor overexpressing mice. AB - The 5-HT3 receptor for serotonin is expressed within limbic structures and is known to modulate neurotransmitter release, suggesting that this receptor may influence learning and memory. Perturbations in serotonergic neurotransmission lead to changes in the ability to attend, learn, and remember. To examine the role of 5-HT3 receptors in learning, memory, and attention, 5-HT3 receptor overexpressing (5-HT3-OE) transgenic mice and their wild-type littermates (WT) were tested in Pavlovian contextual and cued fear conditioning, fear extinction, and latent inhibition (LI) paradigms. Prepulse inhibition (PPI) was assessed to reveal changes in sensorimotor gating. Additionally, anxious behaviors, shock sensitivity, and reactions to novel stimuli were evaluated. 5-HT3-OE mice displayed enhanced contextual conditioning, whereas cued conditioning remained the same as that of WT mice. 5-HT3-OE mice did not differ from WT in extinction rates to either the context or cue. LI was enhanced for 5-HT3-OE mice compared to WT. PPI remained unchanged. No differences in sensitivity to footshock or startle were found. However, 5-HT3-OE mice demonstrated heightened exploratory behavior in response to novel environmental stimuli and decreased anxiety as measured in the elevated plus-maze. Results indicate that overexpression of the 5-HT3 receptor in mouse forebrain results in enhanced hippocampal-dependent learning and attention. Enhanced inspective behavior in response to novelty may contribute to the observed improvements in learning, memory, and attention due to 5-HT3 receptor overexpression. PMID- 14557616 TI - Inability to evoke a long-lasting protective immune response to respiratory syncytial virus infection in mice correlates with ineffective nasal antibody responses. AB - Long-lasting protective antibody is not normally generated in children following primary respiratory syncytial virus (RSV) infection, frequently leading to reinfection. We used the BALB/c mouse model to examine the role of the nasal associated lymphoid tissue and the bone marrow in the generation of RSV-specific long-lasting plasma cells, with a view to further understanding the mechanisms responsible for the poorly sustained RSV antibody levels following primary infection. We show here that substantial numbers of RSV-specific plasma cells were generated in the bone marrow following challenge, which were maintained thereafter. In contrast, in the nasal-associated lymphoid tissue, RSV-specific plasma cell numbers waned quickly both after primary infection and after challenge and were not maintained at a higher level after boosting. These data indicate that the inability to generate a robust local mucosal response in the nasal tissues may contribute substantially to the likelihood of subsequent reinfection and that the presence of serum anti-RSV antibody without local protection is not enough to protect against reinfection. PMID- 14557617 TI - Coronaviruses as vectors: position dependence of foreign gene expression. AB - Coronaviruses are the enveloped, positive-stranded RNA viruses with the largest RNA genomes known. Several features make these viruses attractive as vaccine and therapeutic vectors: (i) deletion of their nonessential genes is strongly attenuating; (ii) the genetic space thus created allows insertion of foreign information; and (iii) their tropism can be modified by manipulation of the viral spike. We studied here their ability to serve as expression vectors by inserting two different foreign genes and evaluating systematically the genomic position dependence of their expression, using a murine coronavirus as a model. Renilla and firefly luciferase expression cassettes, each provided with viral transcription regulatory sequences (TRSs), were inserted at several genomic positions, both independently in different viruses and combined within one viral genome. Recombinant viruses were generated by using a convenient method based on targeted recombination and host cell switching. In all cases high expression levels of the foreign genes were observed without severe effects on viral replication in vitro. The expression of the inserted gene appeared to be dependent on its genomic position, as well as on the identity of the gene. Expression levels increased when the luciferase gene was inserted closer to the 3' end of the genome. The foreign gene insertions generally reduced the expression of upstream viral genes. The results are consistent with coronavirus transcription models in which the transcription from upstream TRSs is attenuated by downstream TRSs. Altogether, our observations clearly demonstrate the potential of coronaviruses as (multivalent) expression vectors. PMID- 14557618 TI - Feline foamy virus genome and replication strategy. AB - Crucial aspects of the foamy virus (FV) replication strategy have so far only been investigated for the prototypic FV (PFV) isolate, which is supposed to be derived from nonhuman primates. To study whether the unusual features of this replication pathway also apply to more-distantly related FVs, we constructed feline FV (FFV) infectious molecular clones and vectors. It is shown by quantitative RNA and DNA PCR analysis that FFV virions contain more RNA than DNA. Full-length linear DNA was found in extracellular FFV by Southern blot analysis. Similar to PFV, azidothymidine inhibition experiments and the transfection of nucleic acids extracted from extracellular FFV indicated that DNA is the functional relevant FFV genome. Unlike PFV, no evidence was found indicating that FFV recycles its DNA into the nucleus. PMID- 14557619 TI - Measles virus (MV) nucleoprotein binds to a novel cell surface receptor distinct from FcgammaRII via its C-terminal domain: role in MV-induced immunosuppression. AB - During acute measles virus (MV) infection, an efficient immune response occurs, followed by a transient but profound immunosuppression. MV nucleoprotein (MV-N) has been reported to induce both cellular and humoral immune responses and paradoxically to account for immunosuppression. Thus far, this latter activity has been attributed to MV-N binding to human and murine FcgammaRII. Here, we show that apoptosis of MV-infected human thymic epithelial cells (TEC) allows the release of MV-N in the extracellular compartment. This extracellular N is then able to bind either to MV-infected or uninfected TEC. We show that recombinant MV N specifically binds to a membrane protein receptor, different from FcgammaRII, highly expressed on the cell surface of TEC. This new receptor is referred to as nucleoprotein receptor (NR). In addition, different Ns from other MV-related morbilliviruses can also bind to FcgammaRII and/or NR. We show that the region of MV-N responsible for binding to NR maps to the C-terminal fragment (N(TAIL)). Binding of MV-N to NR on TEC triggers sustained calcium influx and inhibits spontaneous cell proliferation by arresting cells in the G(0) and G(1) phases of the cell cycle. Finally, MV-N binds to both constitutively expressed NR on a large spectrum of cells from different species and to human activated T cells, leading to suppression of their proliferation. These results provide evidence that MV-N, after release in the extracellular compartment, binds to NR and thereby plays a role in MV-induced immunosuppression. PMID- 14557620 TI - Defining the structure-function relationships of bluetongue virus helicase protein VP6. AB - The VP6 protein of bluetongue virus possesses a number of activities, including nucleoside triphosphatase, RNA binding, and helicase activity (N. Stauber, J. Martinez-Costas, G. Sutton, K. Monastyrskaya, and P. Roy, J. Virol. 71:7220-7226, 1997). Although the enzymatic functions of the protein have been documented, a detailed structure and function study has not been completed and the oligomeric form of the protein in solution has not been described. In this study, we have characterized VP6 activity by creating site-directed mutations in the putative functional helicase domains. Mutant proteins were expressed at high levels in an insect cell by using recombinant baculoviruses purified and analyzed for ATP binding, ATP hydrolysis, and RNA unwinding activities. UV cross-linking experiments indicated that the lysine residue in the conserved motif AXXGXGK(110)V is directly involved in ATP binding, whereas mutant R(205)Q in the arginine-rich motif ER(205)XGRXXR bound ATP at a level comparable to that of the wild-type protein. The RNA binding activity was drastically altered in the R(205)Q mutant and was also affected in the K(110)N mutant. Helicase activity was altered in both mutants. The mutation E(157)N in the DEXX sequence, presumed to act as a Walker B motif, showed an intermediate activity, implying that this motif does not play a crucial role in VP6 function. Purified protein demonstrated stable oligomers with a ring-like morphology in the presence of nucleic acids similar to those shown by other helicases. Gel filtration chromatography, native gel electrophoresis, and glycerol gradient analysis clearly indicated multiple oligomeric forms of VP6. PMID- 14557621 TI - Two distinct size classes of immature and mature subviral particles from tick borne encephalitis virus. AB - Flaviviruses assemble in the endoplasmic reticulum by a mechanism that appears to be driven by lateral interactions between heterodimers of the envelope glycoproteins E and prM. Immature intracellular virus particles are then transported through the secretory pathway and converted to their mature form by cleavage of the prM protein by the cellular protease furin. Earlier studies showed that when the prM and E proteins of tick-borne encephalitis virus are expressed together in mammalian cells, they assemble into membrane-containing, icosahedrally symmetrical recombinant subviral particles (RSPs), which are smaller than whole virions but retain functional properties and undergo cleavage maturation, yielding a mature form in which the E proteins are arranged in a regular T = 1 icosahedral lattice. In this study, we generated immature subviral particles by mutation of the furin recognition site in prM. The mutation resulted in the secretion of two distinct size classes of particles that could be separated by sucrose gradient centrifugation. Electron microscopy showed that the smaller particles were approximately the same size as the previously described mature RSPs, whereas the larger particles were approximately the same size as the virus. Particles of the larger size class were also detected with a wild-type construct that allowed prM cleavage, although in this case the smaller size class was far more prevalent. Subtle differences in endoglycosidase sensitivity patterns suggested that, in contrast to the small particles, the E glycoproteins in the large subviral particles and whole virions might be in nonequivalent structural environments during intracellular transport, with a portion of them inaccessible to cellular glycan processing enzymes. These proteins thus appear to have the intrinsic ability to form alternative assembly products that could provide important clues about the role of lateral envelope protein interactions in flavivirus assembly. PMID- 14557622 TI - Genetically targeted adenovirus vector directed to CD40-expressing cells. AB - The success of gene therapy depends on the specificity of transgene delivery by therapeutic vectors. The present study describes the use of an adenovirus (Ad) fiber replacement strategy for genetic targeting of the virus to human CD40, which is expressed by a variety of diseased tissues. The tropism of the virus was modified by the incorporation into its capsid of a protein chimera comprising structural domains of three different proteins: the Ad serotype 5 fiber, phage T4 fibritin, and the human CD40 ligand (CD40L). The tumor necrosis factor-like domain of CD40L retains its functional tertiary structure upon incorporation into this chimera and allows the virus to use CD40 as a surrogate receptor for cell entry. The ability of the modified Ad vector to infect CD40-positive dendritic cells and tumor cells with a high efficiency makes this virus a prototype of choice for the derivation of therapeutic vectors for the genetic immunization and targeted destruction of tumors. PMID- 14557623 TI - Protein products of the open reading frames encoding nonstructural proteins of human astrovirus serotype 8. AB - Human astroviruses have a positive-strand RNA genome, which contains three open reading frames (ORF1a, ORF1b, and ORF2). The genomic RNA is translated into two nonstructural polyproteins, nsp1a and nsp1ab, that contain sequences derived from ORF1a and from both ORF1a and ORF1b, respectively. Proteins nsp1a and nsp1ab are thought to be proteolytically processed to yield the viral proteins implicated in the replication of the virus genome; however, the intermediate and final products of this processing have been poorly characterized. To identify the cleavage products of the nonstructural polyproteins of a human astrovirus serotype 8 strain, antisera to selected recombinant proteins were produced and were used to analyze the viral proteins synthesized in astrovirus-infected Caco-2 cells and in cells transfected with recombinant plasmids expressing the ORF1a and ORF1b polyproteins. Pulse-chase experiments identified proteins of approximately 145, 88, 85, and 75 kDa as cleavage intermediates during the polyprotein processing. In addition, these experiments and kinetic analysis of the synthesis of the viral proteins identified polypeptides of 57, 20, and 19 kDa, as well as two products of around 27 kDa, as final cleavage products, with the 57-kDa polypeptide most probably being the virus RNA polymerase and the two approximately 27-kDa products being the viral protease. Based on the differential reactivities of the astrovirus proteins with the various antisera used, the individual polypeptides detected were mapped to the virus ORF1a and ORF1b regions. PMID- 14557624 TI - Polyclonal immunoglobulin G from patients neutralizes human immunodeficiency virus type 1 primary isolates by binding free virions, but without interfering with an initial CD4-independent attachment of the virus to primary blood mononuclear cells. AB - We investigated the relationship between human immunodeficiency virus type 1 (HIV 1) primary isolate (PI) antibody-mediated neutralization and attachment to primary blood mononuclear cells (PBMC). Incubation of PIs with immunoglobulin G (IgG) purified from infected patients did not inhibit attachment of the viruses with PBMC, but partial to complete neutralization was achieved. Neutralization of PIs already fixed on the cells was achieved by some IgG samples only and was of limited intensity compared to the former neutralization protocol. On the contrary, the binding of IgG to free virions was shown to be sufficient to reach potent neutralization, as the infectivity of IgG-PI complexes purified from the bulk of antibodies before addition to PBMC was strongly diminished compared to mock-treated controls. Monoclonal antibodies to the CDR2 domain of CD4 completely inhibited the infection of PBMC without interfering with the attachment of PIs to the cells, suggesting that, under these experimental conditions, the initial attachment of viruses to PBMC involves alternative cellular receptors. This initial interaction may also involve other components of the viral envelope than gp120, as partial depletion of the surface glycoproteins of primary viral particles that resulted in an almost complete loss of infectivity did not impair attachment to PBMC. A limited inhibition of attachment was observed when interfering with putative interactions with cellular heparan sulfate, whereas no effect was observed for cellular CD147 or nucleolin or for virion-incorporated cyclophilin A. Altogether, our results favor a mechanism of neutralization of HIV 1 PIs by polyclonal IgG where antibodies predominantly bind free virions and neutralize without interfering with the attachment to PBMC, which, in this model, is mainly CD4 independent. PMID- 14557625 TI - The human immunodeficiency virus type 1 Vif protein reduces intracellular expression and inhibits packaging of APOBEC3G (CEM15), a cellular inhibitor of virus infectivity. AB - Replication of human immunodeficiency virus type 1 (HIV-1) in most primary cells and some immortalized T-cell lines depends on the activity of the viral infectivity factor (Vif). Vif has the ability to counteract a cellular inhibitor, recently identified as CEM15, that blocks infectivity of Vif-defective HIV-1 variants. CEM15 is identical to APOBEC3G and belongs to a family of proteins involved in RNA and DNA deamination. We cloned APOBEC3G from a human kidney cDNA library and confirmed that the protein acts as a potent inhibitor of HIV replication and is sensitive to the activity of Vif. We found that wild-type Vif inhibits packaging of APOBEC3G into virus particles in a dose-dependent manner. In contrast, biologically inactive variants carrying in-frame deletions in various regions of Vif or mutation of two highly conserved cysteine residues did not inhibit packaging of APOBEC3G. Interestingly, expression of APOBEC3G in the presence of wild-type Vif not only affected viral packaging but also reduced its intracellular expression level. This effect was not seen in the presence of biologically inactive Vif variants. Pulse-chase analyses did not reveal a significant difference in the stability of APOBEC3G in the presence or absence of Vif. However, in the presence of Vif, the rate of synthesis of APOBEC3G was slightly reduced. The reduction of intracellular APOBEC3G in the presence of Vif does not fully account for the Vif-induced reduction of virus-associated APOBEC3G, suggesting that Vif may function at several levels to prevent packaging of APOBEC3G into virus particles. PMID- 14557626 TI - Membrane requirements for uridylylation of the poliovirus VPg protein and viral RNA synthesis in vitro. AB - Efficient translation of poliovirus (PV) RNA in uninfected HeLa cell extracts generates all of the viral proteins required to carry out viral RNA replication and encapsidation and to produce infectious virus in vitro. In infected cells, viral RNA replication occurs in ribonucleoprotein complexes associated with clusters of vesicles that are formed from preexisting intracellular organelles, which serve as a scaffold for the viral RNA replication complex. In this study, we have examined the role of membranes in viral RNA replication in vitro. Electron microscopic and biochemical examination of extracts actively engaged in viral RNA replication failed to reveal a significant increase in vesicular membrane structures or the protective aggregation of vesicles observed in PV infected cells. Viral, nonstructural replication proteins, however, bind to heterogeneous membrane fragments in the extract. Treatment of the extracts with nonionic detergents, a membrane-altering inhibitor of fatty acid synthesis (cerulenin), or an inhibitor of intracellular membrane trafficking (brefeldin A) prevents the formation of active replication complexes in vitro, under conditions in which polyprotein synthesis and processing occur normally. Under all three of these conditions, synthesis of uridylylated VPg to form the primer for initiation of viral RNA synthesis, as well as subsequent viral RNA replication, was inhibited. Thus, although organized membranous structures morphologically similar to the vesicles observed in infected cells do not appear to form in vitro, intact membranes are required for viral RNA synthesis, including the first step of forming the uridylylated VPg primer for RNA chain elongation. PMID- 14557627 TI - Binding partners for the UL11 tegument protein of herpes simplex virus type 1. AB - The product of the U(L)11 gene of herpes simplex virus type 1 (HSV-1) is a 96 amino-acid tegument protein that accumulates on the cytoplasmic face of internal membranes. Although it is thought to be important for nucleocapsid envelopment and egress, the actual function of this protein is unknown. Previous studies focused on the characterization of sequence elements within the UL11 protein that function in membrane binding and trafficking to the Golgi apparatus. Binding was found to be mediated by two fatty acyl groups (myristate and palmitate), while an acidic cluster and a dileucine motif were identified as being important for the recycling of UL11 from the plasma membrane to the Golgi apparatus. The goal of the experiments described here was to identify and characterize binding partners (viral or cellular) of UL11. Using both immunoprecipitation and glutathione S transferase (GST) pull-down assays, we identified a 40-kDa protein that specifically associates with UL11 from infected Vero cells. Mutational analyses revealed that the acidic cluster and the dileucine motif are required for this association, whereas the entire second half of UL11 is not. In addition, UL11 homologs from pseudorabies and Marek's disease herpesviruses were also found to be capable of binding to the 40-kDa protein from HSV-1-infected cells, suggesting that the interaction is conserved among alphaherpesviruses. Purification and analysis of the 40-kDa protein by mass spectrometry revealed that it is the product of the U(L)16 gene, a virion protein reported to be involved in nucleocapsid assembly. Cells transfected with a UL16-green fluorescent protein expression vector produced a protein that was of the expected size, could be pulled down with GST-UL11, and accumulated in a Golgi-like compartment only when coexpressed with UL11, indicating that the interaction does not require any other viral products. These data represent the first steps toward elucidating the network of tegument proteins that UL11 links to membranes. PMID- 14557629 TI - Dengue 2 PDK-53 virus as a chimeric carrier for tetravalent dengue vaccine development. AB - Attenuation markers of the candidate dengue 2 (D2) PDK-53 vaccine virus are encoded by mutations that reside outside of the structural gene region of the genome. We engineered nine dengue virus chimeras containing the premembrane (prM) and envelope (E) genes of wild-type D1 16007, D3 16562, or D4 1036 virus within the genetic backgrounds of wild-type D2 16681 virus and the two genetic variants (PDK53-E and PDK53-V) of the D2 PDK-53 vaccine virus. Expression of the heterologous prM-E genes in the genetic backgrounds of the two D2 PDK-53 variants, but not that of wild-type D2 16681 virus, resulted in chimeric viruses that retained PDK-53 characteristic phenotypic markers of attenuation, including small plaque size and temperature sensitivity in LLC-MK(2) cells, limited replication in C6/36 cells, and lack of neurovirulence in newborn ICR mice. Chimeric D2/1, D2/3, and D2/4 viruses replicated efficiently in Vero cells and were immunogenic in AG129 mice. Chimeric D2/1 viruses protected adult AG129 mice against lethal D1 virus challenge. Two tetravalent virus formulations, comprised of either PDK53-E- or PDK53-V-vectored viruses, elicited neutralizing antibody titers in mice against all four dengue serotypes. These antibody titers were similar to the titers elicited by monovalent immunizations, suggesting that viral interference did not occur in recipients of the tetravalent formulations. The results of this study demonstrate that the unique attenuation loci of D2 PDK-53 virus make it an attractive vector for the development of live attenuated flavivirus vaccines. PMID- 14557628 TI - Chromatin remodeling of the Kaposi's sarcoma-associated herpesvirus ORF50 promoter correlates with reactivation from latency. AB - The switch from latent to lytic infection of Kaposi's sarcoma-associated herpesvirus is initiated by the immediate early transcriptional activator protein Rta/open reading frame 50 (ORF50). We examined the transcriptional regulation of the ORF50 core promoter in response to lytic cycle stimulation. We show that the ORF50 promoter is highly responsive to sodium butyrate (NaB) and trichostatin A (TSA), two chemicals known to inhibit histone deacetylases. The NaB and TSA responsive element was mapped to a 70-bp minimal promoter containing an essential GC box that binds Sp1/Sp3 in vitro and in vivo. Micrococcal nuclease mapping studies revealed that a nucleosome is positioned over the transcriptional initiation and the Sp1/3 binding sites. Stimulation with NaB or TSA increased histone acetylation and restriction enzyme accessibility of the ORF50 promoter transcription initiation site. Chromatin immunoprecipitation assay was used to demonstrate that the ORF50 promoter is associated with several different histone deacetylase proteins (including HDAC1, 5, and 7) in latently infected cells. NaB treatment led to the rapid association of Ini1/Snf5, a component of the Swi/Snf family of chromatin remodeling proteins, with the ORF50 promoter. Ectopic expression of the CREB-binding protein (CBP) histone acetyltransferase (HAT) stimulated plasmid-based ORF50 transcription in a HAT-dependent manner, suggesting that CBP recruitment to the ORF50 promoter can be an initiating event for transcription and viral reactivation. Together, these results suggest that remodeling of a stably positioned nucleosome at the transcriptional initiation site of ORF50 is a regulatory step in the transition from latent to lytic infection. PMID- 14557630 TI - Structure and expression of mobile ETnII retroelements and their coding-competent MusD relatives in the mouse. AB - ETnII elements are mobile members of the repetitive early transposon family of mouse long terminal repeat (LTR) retroelements and have caused a number of mutations by inserting into genes. ETnII sequences lack retroviral genes, but the recent discovery of related MusD retroviral elements with regions similar to gag, pro, and pol suggests that MusD provides the proteins necessary for ETnII transposition in trans. For this study, we analyzed all ETnII elements in the draft sequence of the C57BL/6J genome and classified them into three subtypes (alpha, beta, and gamma) based on structural differences. We then used database searches and quantitative real-time PCR to determine the copy number and expression of ETnII and MusD elements in various mouse strains. In 7.5-day-old embryos of a mouse strain in which two mutations due to ETnII-beta insertions have been identified (SELH/Bc), we detected a three- to sixfold higher level of ETnII-beta and MusD transcripts than in control strains (C57BL/6J and LM/Bc). The increased ETnII transcription level can in part be attributed to a higher number of ETnII-beta elements, but 70% of the MusD transcripts appear to have been derived from one or a few MusD elements that are not detectable in C57BL/6J mice. This element belongs to a young MusD subgroup with intact open reading frames and identical LTRs, suggesting that the overexpressed element(s) in SELH/Bc mice might provide the proteins for the retrotransposition of ETnII and MusD elements. We also show that ETnII is expressed up to 30-fold more than MusD, which could explain why only ETnII, but not MusD, elements have been positively identified as new insertions. PMID- 14557631 TI - Development of resistance against diketo derivatives of human immunodeficiency virus type 1 by progressive accumulation of integrase mutations. AB - The diketo acid L-708,906 has been reported to be a selective inhibitor of the strand transfer step of the human immunodeficiency virus type 1 (HIV-1) integration process (D. Hazuda, P. Felock, M. Witmer, A. Wolfe, K. Stillmock, J. A. Grobler, A. Espeseth, L. Gabryelski, W. Schleif, C. Blau, and M. D. Miller, Science 287:646-650, 2000). We have now studied the development of antiviral resistance to L-708,906 by growing HIV-1 strains in the presence of increasing concentrations of the compound. The mutations T66I, L74M, and S230R emerged successively in the integrase gene. The virus with three mutations (T66I L74M S230R) was 10-fold less susceptible to L-708,906, while displaying the sensitivity of the wild-type virus to inhibitors of the RT or PRO or viral entry process. Chimeric HIV-1 strains containing the mutant integrase genes displayed the same resistance profile as the in vitro-selected strains, corroborating the impact of the reported mutations on the resistance phenotype. Phenotypic cross resistance to S-1360, a diketo analogue in clinical trials, was observed for all strains. Interestingly, the diketo acid-resistant strain remained fully sensitive to V-165, a novel integrase inhibitor (C. Pannecouque, W. Pluymers, B. Van Maele, V. Tetz, P. Cherepanov, E. De Clercq, M. Witvrouw, and Z. Debyser, Curr. Biol. 12:1169-1177, 2002). Antiviral resistance was also studied at the level of recombinant integrase. Single mutations did not appear to impair specific enzymatic activity. However, 3' processing and strand transfer activities of the recombinant integrases with two (T66I L74M) and three (T66I L74M S230R) mutations were notably lower than those of the wild-type integrase. Although the virus with three mutations was resistant to inhibition by diketo acids, the sensitivity of the corresponding enzyme to L-708,906 or S-1360 was reduced only two- to threefold. As to the replication kinetics of the selected strains, the replication fitness for all strains was lower than that of the wild-type HIV-1 strain. PMID- 14557632 TI - CpG DNA induces protective antiviral immune responses in Atlantic salmon (Salmo salar L.). AB - Oligodeoxynucleotides (ODN) containing unmethylated CpG dinucleotides within specific sequence contexts (CpG motifs) are detected, like bacterial or viral DNA, as a danger signal by the vertebrate immune system. CpG ODN show promise as vaccine adjuvants and immunoprotective agents in animal models. Here we report that pretreatment with CpG ODN in animals induces nonspecific protection against viral infection. A panel of different synthetic CpG ODN was tested for the in vitro effects in Atlantic salmon (Salmo salar L.) leukocytes. The ODN were tested for their capacity to stimulate proliferation of peripheral blood leukocytes and to induce production of interferon-like factors in head kidney leukocytes. These studies revealed that the sequence and number of the CpG motifs as well as the lengths of the ODN contribute to their stimulatory activity. ODN with the 6-mer CpG motif (5'-GTCGTT-3') showed the highest stimulatory activity and were shown to induce protection against infectious pancreatic necrosis virus when injected in Atlantic salmon. Expression of the Mx transcript, as an indicator of alpha/beta interferon induction, was induced in the CpG-injected fish. These results suggest that CpG DNA in fish induces early, nonspecific antiviral protection. PMID- 14557633 TI - Rescue of the adeno-associated virus genome from a plasmid vector: evidence for rescue by replication. AB - In cultured cells, adeno-associated virus (AAV) replication requires coinfection with a helper virus, either adenovirus or herpesvirus. In the absence of helper virus coinfection AAV can integrate its genome site specifically into the AAVS1 region of chromosome 19. Upon subsequent infection with a helper virus, the AAV genome is released from chromosome 19 by a process termed rescue, and productive replication ensues. The AAV genome cloned into a plasmid vector can also serve to initiate productive AAV replication. When such constructs are transfected into cells and those cells are simultaneously or subsequently infected with a helper virus, the AAV genome is released from the plasmid. This process is thought to serve as a model for rescue from the human genomic site. In this report we present a model for rescue of AAV genomes by replication. A hallmark of this model is the production of a partially single-stranded and partially double stranded molecule. We show that the AAV2 Rep 68 protein, together with the UL30/UL42 herpes simplex virus type 1 DNA polymerase and the UL29 single-strand DNA binding protein ICP8, is sufficient to efficiently and precisely rescue AAV from a plasmid in a way that is dependent on the AAV inverted terminal repeat sequence. PMID- 14557634 TI - Formation of polyomavirus-like particles with different VP1 molecules that bind the urokinase plasminogen activator receptor. AB - Icosahedral virus-like particles formed by the self-assembly of polyomavirus capsid proteins (Py-VLPs) can serve as useful nanostructures for delivering nucleic acids, proteins, and pharmaceuticals into animal cells and tissues. Four predominant surface-exposed loops in the VP1 structure offer potential sites to display sequences that might contribute new targeting specificities. Introduction into each of these loops of sequences derived from the amino-terminal fragment of urokinase plasminogen activator (uPA) or a related phage display peptide reduced the solubility of VP1 molecules when expressed in insect cells, and insertions into the EF loop reduced VP1 solubility least. Coexpression in insect cells of the uPA-VP1 molecules and VP1 containing a FLAG epitope in the HI loop permitted the formation of heterotypic Py-VLPs containing uPA-VP1 and FLAG-VP1. These heterotypic VLPs bound to uPAR on the surfaces of animal cells. Heterotypic Py VLPs containing ligands for multiple cell surface receptors should be useful for targeting specific cells and tissues. PMID- 14557635 TI - Resistance of human cytomegalovirus to the benzimidazole L-ribonucleoside maribavir maps to UL27. AB - 1-(beta-D-Ribofuranosyl)-2,5,6-trichlorobenzimidazole (TCRB) and its 2-bromo analog, BDCRB, are potent and selective inhibitors of human cytomegalovirus (HCMV) DNA processing and packaging. Since they are readily metabolized in vivo, analogs were synthesized to improve biostability. One of these, 1-(beta-L ribofuranosyl)-2-isopropylamino-5,6-dichlorobenzimidazole (1263W94; maribavir), inhibits viral DNA synthesis and nuclear egress. Resistance to maribavir was mapped to UL97, and this viral kinase was shown to be a direct target of maribavir. In the present study, an HCMV strain resistant to TCRB and BDCRB was passaged in increasing concentrations of maribavir, and resistant virus was isolated. This strain (G2) grew at the same rate as the wild-type virus and was resistant to both BDCRB and maribavir. Resistance to BDCRB was expected, because the parent strain from which G2 was isolated was resistant due to known mutations in UL56 and UL89. However, no mutations were found in UL97 or other relevant open reading frames that could explain resistance to maribavir. Because sequencing of selected HCMV genes did not identify the resistance mutation, a cosmid library was made from G2, and a series of recombinant G2 wild-type viruses were constructed. Testing the recombinants for sensitivity to maribavir narrowed the locus of resistance to genes UL26 to UL32. Sequencing identified a single coding mutation in ORF UL27 (Leu335Pro) as the one responsible for resistance to maribavir. These results establish that UL27 is either directly or indirectly involved in the mechanism of action of maribavir. They also suggest that UL27 could play a role in HCMV DNA synthesis or egress of HCMV particles from the nucleus. PMID- 14557636 TI - HLA-A11-restricted epitope polymorphism among Epstein-Barr virus strains in the highly HLA-A11-positive Chinese population: incidence and immunogenicity of variant epitope sequences. AB - An individual's CD8(+)-cytotoxic-T-lymphocyte (CTL) response to Epstein-Barr virus (EBV) latent cycle antigens focuses on a small number of immunodominant epitopes often presented by just one of the available HLA class I alleles; for example, HLA-A11-positive Caucasians frequently respond to two immunodominant HLA A11 epitopes, IVTDFSVIK (IVT) and AVFDRKSDAK (AVF), within the nuclear antigen EBNA3B. Here, we reexamine the spectrum of EBV strains present in the highly HLA A11-positive Chinese population for sequence changes in these epitopes relative to the Caucasian type 1 prototype strain B95.8. The IVT epitope was altered in 61 of 64 Chinese type 1 viruses, with four different sequence variants being observed, and the AVF epitope was altered in 46 cases with six different sequence variants; by contrast, all 10 Chinese type 2 viruses retained the prototype 2 epitope sequences. All but one of the type 1 epitope variants were poorly recognized by IVT- or AVF-specific CTLs in pulse-chase assays of peptide-mediated target cell lysis. More importantly, we screened HLA-A11-positive Chinese donors carrying viruses with known epitope mutations for evidence of epitope-specific CTL memory by enzyme-linked immunospot assays: none of the type 1 variants tested, nor the type 2 prototype, appeared to be immunogenic in vivo. The data remain consistent with the possibility that, during virus-host coevolution, pressure from the host CTL-mediated immune response has given A11 epitope-loss viruses a selective advantage. PMID- 14557637 TI - Latent gene sequencing reveals familial relationships among Chinese Epstein-Barr virus strains and evidence for positive selection of A11 epitope changes. AB - Epstein-Barr virus (EBV) strains from the highly HLA-A11-positive Chinese population are predominantly type 1 and show a variety of sequence changes (relative to the contemporary Caucasian prototype strain B95.8) in the nuclear antigen EBNA3B sequences encoding two immunodominant HLA-A11 epitopes, here called IVT and AVF. This has been interpreted by some as evidence of immune selection and by others as random genetic drift. To study epitope variation in a broader genomic context, we sequenced the whole of EBNA3B and parts of the EBNA2, 3A, and 3C genes from each of 31 Chinese EBV isolates. At each locus, type 1 viruses showed <2% nucleotide divergence from the B95.8 prototype while type 2 sequences remained even closer to the contemporary African prototype Ag876. However, type 1 isolates could clearly be divided into families based on linked patterns of sequence divergence from B95.8 across all four EBNA loci. Different patterns of IVT and AVF variation were associated with the different type 1 families, and there was additional epitope diversity within families. When the EBNA3 gene sequences of type 1 Chinese strains were subject to computer-based analysis, particular codons within the A11-epitope-coding region were among the few identified as being under positive or diversifying selection pressure. From these results, and the observation that mutant epitopes are consistently nonimmunogenic in vivo, we conclude that the immune selection hypothesis remains viable and worthy of further investigation. PMID- 14557638 TI - Human immunodeficiency virus type 1 escape from RNA interference. AB - Sequence-specific degradation of mRNA by short interfering RNA (siRNA) allows the selective inhibition of viral proteins that are critical for human immunodeficiency virus type 1 (HIV-1) replication. The aim of this study was to characterize the potency and durability of virus-specific RNA interference (RNAi) in cell lines that stably express short hairpin RNA (shRNA) targeting the HIV-1 transactivator protein gene tat. We found that the antiviral activity of tat shRNA was abolished due to the emergence of viral quasispecies harboring a point mutation in the shRNA target region. Our results suggest that, in order for RNAi to durably suppress HIV-1 replication, it may be necessary to target highly conserved regions of the viral genome. Alternatively, similar to present antiviral drug therapy paradigms, DNA constructs expressing multiple siRNAs need to be developed that target different regions of the viral genome, thereby reducing the probability of generating escape mutants. PMID- 14557639 TI - CD4 and major histocompatibility complex class I downregulation by the human immunodeficiency virus type 1 nef protein in pediatric AIDS progression. AB - The human immunodeficiency virus type 1 (HIV-1) nef gene is a crucial determinant in AIDS disease progression. Although several in vitro activities have been attributed to the Nef protein, identifying the one critical for in vivo pathogenicity remains elusive. In this study, we examined a large number of nef alleles derived at various time points from 13 perinatally infected children showing different progression rates: six nonprogressors (NPs), three slow progressors (SPs), and four rapid progressors (RPs). The patient-derived nef alleles were analyzed for their steady-state expression of a Nef protein, for their relative ability to downregulate cell surface expression of CD4 and major histocompatibility complex class I (MHC-I) and for their capacity to bind the clathrin adaptor AP-1 complex. We found that NP-derived nef alleles, compared to nef alleles isolated from SPs and RPs, had reduced CD4 and MHC-I downregulation activities. In contrast, SP- and RP-derived nef alleles did not differ and efficiently downregulated both CD4 and MHC-I. AP-1 binding was a conserved function of primary nef alleles not correlated with clinical progression. Defective Nef proteins from NPs, rather than sharing common specific changes in their sequences, accumulated various amino acid substitutions, mainly located outside the conserved domains previously associated with Nef biological properties. Our data indicate that Nef-mediated downregulation of cell surface CD4 and MHC-I significantly contributes to the expression of the pathogenic potential of HIV-1. PMID- 14557641 TI - Expression of the zinc-finger antiviral protein inhibits alphavirus replication. AB - The rat zinc-finger antiviral protein (ZAP) was recently identified as a host protein conferring resistance to retroviral infection. We analyzed ZAP's ability to inhibit viruses from other families and found that ZAP potently inhibits the replication of multiple members of the Alphavirus genus within the Togaviridae, including Sindbis virus, Semliki Forest virus, Ross River virus, and Venezuelan equine encephalitis virus. However, expression of ZAP did not induce a broad spectrum antiviral state as some viruses, including vesicular stomatitis virus, poliovirus, yellow fever virus, and herpes simplex virus type 1, replicated to normal levels in ZAP-expressing cells. We determined that ZAP expression inhibits Sindbis virus replication after virus penetration and entry, but before the amplification of newly synthesized plus strand genomic RNA. Using a temperature sensitive Sindbis virus mutant expressing luciferase, we further showed that translation of incoming viral RNA is blocked by ZAP expression. Elucidation of the antiviral mechanism by which ZAP inhibits Sindbis virus translation may lead to the development of agents with broad activity against alphaviruses. PMID- 14557640 TI - A molecularly cloned Schwarz strain of measles virus vaccine induces strong immune responses in macaques and transgenic mice. AB - Live attenuated RNA viruses make highly efficient vaccines. Among them, measles virus (MV) vaccine has been given to a very large number of children and has been shown to be highly efficacious and safe. Therefore, this vaccine might be a very promising vector to immunize children against both measles and other infectious agents, such as human immunodeficiency virus. A vector was previously derived from the Edmonston B strain of MV, a vaccine strain abandoned 25 years ago. Sequence analysis revealed that the genome of this vector diverges from Edmonston B by 10 amino acid substitutions not related to any Edmonston subgroup. Here we describe an infectious cDNA for the Schwarz/Moraten strain, a widely used MV vaccine. This cDNA was constructed from a batch of commercial vaccine. The extremities of the cDNA were engineered in order to maximize virus yield during rescue. A previously described helper cell-based rescue system was adapted by cocultivating transfected cells on primary chicken embryo fibroblasts, the cells used to produce the Schwarz/Moraten vaccine. After two passages the sequence of the rescued virus was identical to that of the cDNA and of the published Schwarz/Moraten sequence. Two additional transcription units were introduced in the cDNA for cloning foreign genetic material. The immunogenicity of rescued virus was studied in macaques and in mice transgenic for the CD46 MV receptor. Antibody titers and T-cell responses (ELISpot) in animals inoculated with low doses of rescued virus were identical to those obtained with commercial Schwarz MV vaccine. In contrast, the immunogenicity of the previously described Edmonston B strain-derived MV clone was much lower. This new molecular clone will allow for the production of MV vaccine without having to rely on seed stocks. The additional transcription units allow expressing heterologous antigens, thereby providing polyvalent vaccines based on an approved, safe, and efficient MV vaccine strain that is used worldwide. PMID- 14557643 TI - Caprine arthritis-encephalitis virus envelope surface glycoprotein regions interacting with the transmembrane glycoprotein: structural and functional parallels with human immunodeficiency virus type 1 gp120. AB - A sequence similarity between surface envelope glycoprotein (SU) gp135 of the lentiviruses maedi-visna virus and caprine arthritis-encephalitis virus (CAEV) and human immunodeficiency virus type 1 (HIV-1) gp120 has been described. The regions of sequence similarity are in the second and fifth conserved regions of gp120, and the similarity is highest in sequences coinciding with beta-strands 4 to 8 and 25, which are located in the most virion-proximal region of the gp120 inner domain. A subset of this structure, formed by gp120 beta-strands 4, 5, and 25, is conserved in most or all lentiviruses. Because of the orientation of gp120 on the virion, this highly conserved virion-proximal region of the gp120 core may interact with the transmembrane glycoprotein (TM) together with the amino and carboxy termini of full-length gp120. Therefore, interactions between SU and TM of lentiviruses may be structurally related. Here we tested whether the amino acid residues in the putative virion-proximal region of CAEV gp135 comprising putative beta-strands 4, 5, and 25, as well as its amino and carboxy termini, are important for stable interactions with TM. An amino acid change at gp135 position 119 or 521, located in the turn between putative beta-strands 4 and 5 and near beta-strand 25, respectively, specifically disrupted the epitope recognized by monoclonal antibody 29A. Thus, similar to the corresponding gp120 regions, these gp135 residues are located in close proximity to each other in the folded protein, supporting the hypothesis of a structural similarity between the gp120 virion-proximal inner domain and gp135. Amino acid changes in the amino- and carboxy-terminal and putative virion-proximal regions of gp135 increased gp135 shedding from the cell surface, indicating that these gp135 regions are involved in interactions with TM. Our results indicate structural and functional parallels between CAEV gp135 and HIV-1 gp120 that may be more broadly applicable to the SU of other lentiviruses. PMID- 14557642 TI - Multigene DNA priming-boosting vaccines protect macaques from acute CD4+-T-cell depletion after simian-human immunodeficiency virus SHIV89.6P mucosal challenge. AB - We evaluated four priming-boosting vaccine regimens for the highly pathogenic simian human immunodeficiency virus SHIV89.6P in Macaca nemestrina. Each regimen included gene gun delivery of a DNA vaccine expressing all SHIV89.6 genes plus Env gp160 of SHIV89.6P. Additional components were two recombinant vaccinia viruses, expressing SHIV89.6 Gag-Pol or Env gp160, and inactivated SHIV89.6 virus. We compared (i) DNA priming/DNA boosting, (ii) DNA priming/inactivated virus boosting, (iii) DNA priming/vaccinia virus boosting, and (iv) vaccinia virus priming/DNA boosting versus sham vaccines in groups of 6 macaques. Prechallenge antibody responses to Env and Gag were strongest in the groups that received vaccinia virus priming or boosting. Cellular immunity to SHIV89.6 peptides was measured by enzyme-linked immunospot assay; strong responses to Gag and Env were found in 9 of 12 vaccinia virus vaccinees and 1 of 6 DNA primed/inactivated-virus-boosted animals. Vaccinated macaques were challenged intrarectally with 50 50% animal infectious doses of SHIV89.6P 3 weeks after the last immunization. All animals became infected. Five of six DNA-vaccinated and 5 of 6 DNA-primed/particle-boosted animals, as well as all 6 controls, experienced severe CD4(+)-T-cell loss in the first 3 weeks after infection. In contrast, DNA priming/vaccinia virus boosting and vaccinia virus priming/DNA boosting vaccines both protected animals from disease: 11 of 12 macaques had no loss of CD4(+) T cells or moderate declines. Virus loads in plasma at the set point were significantly lower in vaccinia virus-primed/DNA-boosted animals versus controls (P = 0.03). We conclude that multigene vaccines delivered by a combination of vaccinia virus and gene gun-delivered DNA were effective against SHIV89.6P viral challenge in M. nemestrina. PMID- 14557644 TI - Replication of an E1B 55-kilodalton protein-deficient adenovirus (ONYX-015) is restored by gain-of-function rather than loss-of-function p53 mutants. AB - ONYX-015 (dl1520) is an E1B 55-kilodalton protein-deficient replicating adenovirus that is currently in clinical trials as an antitumor agent. On the basis of the observation that the E1B 55kD gene product is able to bind to and inactivate p53, ONYX-015's mechanism of action is proposed to involve selective replication in and killing of p53-deficient cells. While its efficacy as a therapeutic agent appears evident, the virus's mechanism of cellular selectivity, including a possible role of p53 in this regard, is less clear. Indeed, there have been a number of recent reports suggesting that the p53 status of target cells does not reliably predict ONYX-015 replication or cell killing. To address the role of p53 in ONYX-015 selectivity, we have undertaken a rigorous analysis of the behavior of this virus in small airway-derived primary human epithelial cells expressing either dominant-negative or gain-of-function mutant p53 genes. Examination of small airway epithelial cells expressing a variety of p53 mutant alleles revealed that while all were able to inhibit endogenous p53 activity, only one allele examined, 248W, demonstrated a markedly increased ability to facilitate ONYX-015 replication. This allele is a member of a group of p53 mutants (know as class I mutants) characterized by retention of global structural conformation but loss of DNA-binding activity. These observations indicate that the nature of the p53 mutation affects ONYX-015 replication, help reconcile disparate published findings, and may provide criteria by which to direct clinical application of ONYX-015. PMID- 14557645 TI - Optimal induction of T-cell responses against hepatitis C virus E2 by antigen engineering in DNA immunization. AB - Although DNA immunization is a safe and efficient method for inducing cellular immune responses, it generates relatively weak and slow immune responses. Here, we investigated the effect of hepatitis C virus (HCV) antigen modifications on the induction of T-cell responses in DNA immunization. It is likely that the strength of T-cell responses has an inverse relationship with the length of the insert DNA. Interestingly, a mixture of several plasmids carrying each gene induced a higher level of T-cell responses than a single plasmid expressing a long polyprotein. Moreover, the presence of a transmembrane domain in HCV E2 resulted in stronger T-cell responses against E2 protein than its absence. Taken together, our results indicate that the tailored modifications of DNA-encoded antigens are capable of optimizing the induction of T-cell responses which is required for eliminating the cells chronically infected with highly variable viruses such as HCV and human immunodeficiency virus. PMID- 14557646 TI - The 3' end of Norwalk virus mRNA contains determinants that regulate the expression and stability of the viral capsid protein VP1: a novel function for the VP2 protein. AB - Norwalk virus (NV) is the prototype strain of a group of noncultivable human caliciviruses responsible for epidemic outbreaks of acute gastroenteritis. The capsid protein VP1 is synthesized from a subgenomic RNA that contains two open reading frames (ORFs), ORF2 and ORF3, and the 3' untranslated region (UTR). ORF2 and ORF3 code for the capsid protein (VP1) and a small structural basic protein (VP2), respectively. We discovered that the yields of virus-like particles (VLPs) composed of VP1 are significantly reduced when this protein is expressed from ORF2 alone. To determine how the 3' terminus of the NV subgenomic RNA regulates VP1 expression, we compared VP1 expression levels by using recombinant baculovirus constructs containing different 3' elements. High VP1 levels were detected by using a recombinant baculovirus that contained ORF2, ORF3, and the 3'UTR (ORF2+3+3'UTR). In contrast, expression of VP1 from constructs that lacked the 3'UTR (ORF2+3), ORF3 (ORF2+3'UTR), or both (ORF2 alone) was highly reduced. Elimination of VP2 synthesis from the subgenomic RNA by mutation resulted in VP1 levels similar to those obtained with the ORF2 construct alone, suggesting a cis role for VP2 in upregulation of VP1 expression levels. Comparisons of the kinetics of RNA and capsid protein expression levels by using constructs with or without ORF3 or the 3'UTR revealed that the 3'UTR increased the levels of VP1 RNA, whereas the presence of VP2 resulted in increased levels of VP1. Furthermore, VP2 increased VP1 stability and protected VP1 from disassembly and protease degradation. The increase in VP1 expression levels caused by the presence of VP2 in cis was also observed in mammalian cells. PMID- 14557648 TI - Identification of a human papillomavirus type 16-specific epitope on the C terminal arm of the major capsid protein L1. AB - To characterize epitopes on human papillomavirus (HPV) virus-like particles (VLPs), a panel of mutated HPV-16 VLPs was created. Each mutated VLP had residues substituted from HPV-31 or HPV-52 L1 sequences to the HPV-16 L1 backbone. Mutations were created on the HPV-31 and -52 L1 proteins to determine if HPV-16 type-specific recognition could be transferred. Correct folding of the mutated proteins was verified by resistance to trypsin digestion and by binding to one or more conformation-dependent monoclonal antibodies. Several of the antibodies tested were found to bind to regions already identified as being important for HPV VLP recognition (loops DE, EF, FG, and HI). Sequences at both ends of the long FG loop (amino acids 260 to 290) were required for both H16.V5 and H16.E70 reactivity. A new antibody-binding site was discovered on the C-terminal arm of L1 between positions 427 and 445. Recognition of these residues by the H16.U4 antibody suggests that this region is surface exposed and supports a recently proposed molecular model of HPV VLPs. PMID- 14557647 TI - Changes in mumps virus gene sequence associated with variability in neurovirulent phenotype. AB - Mumps virus is highly neurotropic and, prior to widespread vaccination programs, was the major cause of viral meningitis in the United States. Nonetheless, the genetic basis of mumps virus neurotropism and neurovirulence was until recently not understood, largely due to the lack of an animal model. Here, nonneurovirulent (Jeryl Lynn vaccine) and highly neurovirulent (88-1961 wild type) mumps virus strains were passaged in human neural cells or in chicken fibroblast cells with the goal of neuroadapting or neuroattenuating the viruses, respectively. When tested in our rat neurovirulence assay against the respective parental strains, a Jeryl Lynn virus variant with an enhanced propensity for replication (neurotropism) and damage (neurovirulence) in the brain and an 88 1961 wild-type virus variant with decreased neurotropic and neurovirulent properties were recovered. To determine the molecular basis for the observed differences in neurovirulence and neuroattenuation, the complete genomes of the parental strains and their variants were fully sequenced. A comparison at the nucleotide level associated three amino acid changes with enhanced neurovirulence of the neuroadapted vaccine strain: one each in the nucleoprotein, matrix protein, and polymerase and three amino acid changes with reduced neurovirulence of the neuroattenuated wild-type strain: one each in the fusion protein, hemagglutinin-neuraminidase protein, and polymerase. The potential role of these amino acid changes in neurotropism, neurovirulence, and neuroattenuation is discussed. PMID- 14557649 TI - Murine cytomegalovirus m41 open reading frame encodes a Golgi-localized antiapoptotic protein. AB - Viruses have evolved various strategies to prevent premature apoptosis of infected host cells. Some of the viral genes mediating antiapoptotic functions have been identified by their homology to cellular genes, but others are structurally unrelated to genes of known function. In this study, we used a random, unbiased approach to identify such genes in the murine cytomegalovirus genome. From a library of random transposon insertion mutants, a mutant virus that caused premature cell death was isolated. The transposon was inserted within open reading frame m41. An independently constructed m41 deletion mutant showed the same phenotype, whereas deletion mutants lacking the adjacent genes m40 and M42 did not. Apoptosis occurred in different cell types, could be blocked by caspase inhibitors, and did not require p53. Within the murine cytomegalovirus genome, m41, m40, and m39 form a small cluster of genes of unknown function. They are homologous to r41, r40, and r39 of rat cytomegalovirus, but lack sequence homology to UL41, UL40, and UL37 exon 1 (UL37x1) which are located at the corresponding positions of the human cytomegalovirus genome. Unlike UL37x1 of human cytomegalovirus, which encodes a mitochondrion-localized inhibitor of apoptosis that is essential for virus replication, m41 encodes a protein that localizes to the Golgi apparatus. The murine cytomegalovirus m41 product is the first example of a Golgi-localized protein that prevents premature apoptosis and thus extends the life span of infected cells. PMID- 14557650 TI - Cell surface expression of major histocompatibility complex class I molecules is reduced in hepatitis C virus subgenomic replicon-expressing cells. AB - The hepatitis C virus (HCV) causes chronic hepatitis in most infected individuals by evading host immune defenses. In this investigation, we show that HCV-infected cells may go undetected in the immune system by suppressing major histocompatibility complex (MHC) class I antigen presentation to cytotoxic T lymphocytes. Cells expressing HCV subgenomic replicons have lower MHC class I cell surface expression. This is due to reduced levels of properly folded MHC class I molecules. HCV replicons induce endoplasmic reticulum (ER) stress (K. Tardif, K. Mori, and A. Siddiqui, J. Virol. 76:7453-7459, 2002), which results from a decline in protein glycosylation. Decreasing protein glycosylation can disrupt protein folding, preventing the assembly of MHC class I molecules. This results in the accumulation of unfolded MHC class I. Therefore, the persistence and pathogenesis of HCV may depend upon the ER stress-mediated interference of MHC class I assembly and cell surface expression. PMID- 14557651 TI - Murine leukemia virus particle assembly quantitated by fluorescence microscopy: role of Gag-Gag interactions and membrane association. AB - In order to track the assembly of murine leukemia virus (MLV), we used fluorescence microscopy to visualize particles containing Gag molecules fused to fluorescent proteins (FPs). Gag-FP chimeras budded from cells to produce fluorescent spots, which passed through the same pore-size filters and sedimented at the same velocity as authentic MLV. N-terminal myristylation of Gag-FPs was necessary for particle formation unless wild-type Gag was coexpressed. By labeling nonmyristylated Gag with yellow FP and wild-type Gag with cyan FP, we could quantitate the coincorporation of two proteins into single particles. This experiment showed that nonmyristylated Gag was incorporated into mixed particles at approximately 50% the efficiency of wild-type Gag. Mutations that inhibit Gag Gag interactions (K. Alin and S. P. Goff, Virology 216:418-424, 1996; K. Alin and S. P. Goff, Virology 222:339-351, 1996) were then introduced into the capsid (CA) region of Gag-FPs. The mutations P150L and R119C/P133L inhibited fluorescent particle formation by these Gag-FPs, but Gag-FPs containing these mutations could be efficiently incorporated into particles when coexpressed with wild-type Gag. When these mutations were introduced into nonmyristylated Gag-FPs, no incorporation into particles in the presence of wild-type Gag was detected. These data suggest that two independent mechanisms, CA interactions and membrane association following myristylation, cooperate in MLV Gag assembly and budding. PMID- 14557653 TI - Mutational inactivation of two distinct negative RNA elements in the human papillomavirus type 16 L2 coding region induces production of high levels of L2 in human cells. AB - Here we show that the 5' end and the middle region of the L2 coding sequence of human papillomavirus type 16 contain strong inhibitory RNA sequences termed inhibitory regions I and II. This is in contrast to L1, which contains one inhibitory region in the 5' end of the coding region. Inhibitory regions I and II acted in cis to reduce L2 mRNA levels and to inhibit the use of the mRNA. In tandem, the two regions reduced L2 mRNA production to undetectable levels. Specific mutational inactivation of the two inhibitory elements in the 5' end and in the middle region of L2 by the introduction of nucleotide substitutions that changed the nucleotide sequence but not the protein sequence resulted in production of high levels of L2 mRNA and protein. In contrast to L2, a partial L1 mutant in which only the first one third of L1 was mutated produced levels of L1 mRNA and protein similar to those in a full L1 mutant. In addition, the constitutive transport element of simian retrovirus type 1 overcomes the effect of the inhibitory sequences of L1 but not L2. PMID- 14557652 TI - A locus on mouse chromosome 6 that determines resistance to herpes simplex virus also influences reactivation, while an unlinked locus augments resistance of female mice. AB - During studies to determine a role for tumor necrosis factor (TNF) in herpes simplex virus type 1 (HSV-1) infection using TNF receptor null mutant mice, we discovered a genetic locus, closely linked to the TNF p55 receptor (Tnfrsf1a) gene on mouse chromosome 6 (c6), that determines resistance or susceptibility to HSV-1. We named this locus the herpes resistance locus, Hrl, and showed that it also mediates resistance to HSV-2. Hrl has at least two alleles, Hrl(r), expressed by resistant strains like C57BL/6 (B6), and Hrl(s), expressed by susceptible strains like 129S6 (129) and BALB/c. Although Hrl is inherited as an autosomal dominant gene, resistance to HSV-1 is strongly sex biased such that female mice are significantly more resistant than male mice. Analysis of backcrosses between resistant B6 and susceptible 129 mice revealed that a second locus, tentatively named the sex modifier locus, Sml, functions to augment resistance of female mice. Besides determining resistance, Hrl is one of several genes involved in the control of HSV-1 replication in the eye and ganglion. Remarkably, Hrl also affects reactivation of HSV-1, possibly by interaction with some unknown gene(s). We showed that Hrl is distinct from Cmv1, the gene that determines resistance to murine cytomegalovirus, which is encoded in the major NK cell complex just distal of p55 on c6. Hrl has been mapped to a roughly 5 centimorgan interval on c6, and current efforts are focused on obtaining a high resolution map for Hrl. PMID- 14557654 TI - Distinct domains in the adenovirus E3 RIDalpha protein are required for degradation of Fas and the epidermal growth factor receptor. AB - Adenovirus (Ad) types 2 and 5 encode at least five proteins within the E3 transcription unit that help the virus evade the immune system. Two such proteins, RIDalpha (formerly E3-10.4K) and RIDbeta (formerly E3-14.5K), form the RID (receptor internalization and degradation) complex (formerly E3-10.4K/14.5K). RID mediates clearance from the cell surface and lysosomal degradation of a number of important members in the tumor necrosis factor receptor (TNFR) superfamily and the receptor tyrosine kinase receptor family. Affected receptors include Fas, TRAIL (TNF-related apoptosis-inducing ligand) receptor 1 (TR1), TR2, and epidermal growth factor receptor (EGFR). Degradation of Fas and TRAIL receptors protects Ad-infected cells from apoptosis. To investigate the mechanism of action of RIDalpha, 14 mutant RIDalpha proteins, each containing a three- to five-amino-acid deletion, were constructed and then expressed from the E3 region of a replication-competent recombinant Ad in the same context as wild-type RIDalpha. Each mutant protein was characterized with regard to five physical properties associated with wild-type RIDalpha, namely, protein stability, proteolytic cleavage, insertion into the membrane, complex formation with RIDbeta, and transport to the cell surface. Additionally, the mutant proteins were tested for their ability to mediate internalization and degradation of EGFR and Fas and to protect cells from Fas-mediated apoptosis. The majority of mutant RIDalpha proteins (8 out of 14) were physically similar to wild-type RIDalpha. With regard to functional characteristics, the cytoplasmic domain of RIDalpha is largely unimportant for receptor internalization and degradation and the extracellular domain of RIDalpha is important for down-regulation of EGFR but not Fas. PMID- 14557655 TI - Hypovirus papain-like protease p29 functions in trans to enhance viral double stranded RNA accumulation and vertical transmission. AB - The prototypic hypovirus CHV1-EP713 attenuates virulence (hypovirulence) and alters several physiological processes of the chestnut blight fungus Cryphonectria parasitica. The papain-like protease, p29, and the highly basic protein, p40, derived, respectively, from the N-terminal and C-terminal portions of the CHV1-EP713-encoded open reading frame (ORF) A polyprotein, p69, both contribute to reduced pigmentation and sporulation. The p29 coding region was shown to suppress pigmentation and asexual sporulation in the absence of virus infection in transformed C. parasitica, whereas transformants containing the p40 coding domain exhibited a wild-type, untransformed phenotype. Deletion of either p29 or p40 from the viral genome also results in reduced accumulation of viral RNA. We now show that p29, but not p40, functions in trans to enhance genomic RNA accumulation and vertical transmission of p29 deletion mutant viruses. The frequency of virus transmission through conidia was found to decrease with reduced accumulation of viral genomic double-stranded RNA (dsRNA): from almost 100% for wild-type virus to approximately 50% for Deltap29, and 10 to 20% for Deltap69. When expressed from a chromosomally integrated cDNA copy, p29 elevated viral dsRNA accumulation and transmission for Deltap29 mutant virus to the level shown by wild-type virus. Increased viral RNA accumulation levels were also observed for a Deltap69 mutant lacking almost the entire ORF A sequence. Such enhancements were not detected in transgenic fungal colonies expressing p40. Mutation of p29 residues Cys(70) or Cys(72), strictly conserved in hypovirus p29 and potyvirus HC-Pro, resulted in the loss of both p29-mediated suppressive activity in virus-free transgenic C. parasitica and in trans enhancement of RNA accumulation and transmission, suggesting a linkage between these functional activities. These results suggest that p29 is an enhancer of viral dsRNA accumulation and vertical virus transmission through asexual spores. PMID- 14557656 TI - Severe CD4+ T-cell depletion in gut lymphoid tissue during primary human immunodeficiency virus type 1 infection and substantial delay in restoration following highly active antiretroviral therapy. AB - Gut-associated lymphoid tissue (GALT) harbors the majority of T lymphocytes in the body and is an important target for human immunodeficiency virus type 1 (HIV 1). We analyzed longitudinal jejunal biopsy samples from HIV-1-infected patients, during both primary and chronic stages of HIV-1 infection, prior to and following the initiation of highly active antiretroviral therapy (HAART) to determine the onset of CD4(+) T-cell depletion and the effect of HAART on the restoration of CD4(+) T cells in GALT. Severe depletion of intestinal CD4(+) T cells occurred during primary HIV-1 infection. Our results showed that the restoration of intestinal CD4(+) T cells following HAART in chronically HIV-1-infected patients was substantially delayed and incomplete. In contrast, initiation of HAART during early stages of infection resulted in near-complete restoration of intestinal CD4(+) T cells, despite the delay in comparison to peripheral blood CD4(+) T-cell recovery. DNA microarray analysis of gene expression profiles and flow-cytometric analysis of lymphocyte homing and cell proliferation markers demonstrated that cell trafficking to GALT and not local proliferation contributed to CD4(+) T-cell restoration. Evaluation of jejunal biopsy samples from long-term HIV-1-infected nonprogressors showed maintenance of normal CD4(+) T-cell levels in both GALT and peripheral blood. Our results demonstrate that near-complete restoration of mucosal immune system can be achieved by initiating HAART early in HIV-1 infection. Monitoring of the restoration and/or maintenance of CD4(+) T cells in GALT provides a more accurate assessment of the efficacy of antiviral host immune responses as well as HAART. PMID- 14557657 TI - Array analysis of viral gene transcription during lytic infection of cells in tissue culture with Varicella-Zoster virus. AB - Varicella-zoster virus (VZV), a neurotropic alphaherpesvirus, causes childhood chickenpox (varicella), becomes latent in dorsal root and autonomic ganglia, and reactivates decades later to cause shingles (zoster) and other neurologic complications. Although the sequence and configuration of VZV DNA have been determined, relatively little is known about viral gene expression in productively infected cells. This is in part because VZV is highly cell associated, and sufficient titers of cell-free virus for use in synchronizing infection do not develop. PCR-based transcriptional arrays were constructed to simultaneously determine the relative abundance of the approximately 70 predicted VZV open reading frames (ORFs). Fragments (250 to 600 bp) from the 5' and 3' end of each ORF were PCR amplified and inserted into plasmid vectors. The virus DNA inserts were amplified, quantitated, and spotted onto nylon membranes. Probing the arrays with radiolabeled cDNA synthesized from VZV-infected cells revealed an increase in the magnitude of the expressed VZV genes from days 1 to 3 after low multiplicity virus infection but little change in their relative abundance. The most abundant VZV transcripts mapped to ORFs 9/9A, 64, 33/33A, and 49, of which only ORF 9 corresponded to a previously identified structural gene. Array analysis also mapped transcripts to three large intergenic regions previously thought to be transcriptionally silent, results subsequently confirmed by Northern blot and reverse transcription-PCR analysis. Array analysis provides a formidable tool to analyze transcription of an important ubiquitous human pathogen. PMID- 14557659 TI - Oligodendrocyte-specific expression of human immunodeficiency virus type 1 Nef in transgenic mice leads to vacuolar myelopathy and alters oligodendrocyte phenotype in vitro. AB - Vacuolar myelopathy (VM) is a frequent central nervous system complication of human immunodeficiency virus type 1 (HIV-1) infection. We report here that transgenic (Tg) mice expressing even low levels of Nef in oligodendrocytes under the regulation of the myelin basic protein (MBP) promoter (MBP/HIV(Nef)) developed VM similar to the human disease in its appearance and topography. The spinal cords of these Tg mice showed lower levels of the myelin proteins MAG and CNPase and of the 21-kDa isoform of MBP prior to the development of vacuoles. In addition, Tg oligodendrocytes in primary in vitro cultures appeared morphologically more mature but, paradoxically, exhibited a less mature phenotype based on O4, O1, CNPase, and MBP staining. In particular, mature CNPase(+) MBP(+) Tg oligodendrocytes were less numerous than non-Tg oligodendrocytes. Therefore, Nef appears to affect the proper differentiation of oligodendrocytes. These data suggest that even low levels of Nef expression in human oligodendrocytes may be responsible for the development of VM in HIV-1-infected individuals. PMID- 14557658 TI - The AIDS-like disease of CD4C/human immunodeficiency virus transgenic mice is associated with accumulation of immature CD11bHi dendritic cells. AB - CD4C/human immunodeficiency virus (HIV) transgenic mice develop an AIDS-like disease. We used this model to study the effects of HIV-1 on dendritic cells (DC). We found a progressive decrease in total DC numbers in the lymph nodes, with a significant accumulation of CD11b(Hi) DC. In the thymus, the recovery of transgenic CD8alpha(+) DC had a tendency to be lower. Spleen DC were augmented in the marginal zone. Transgenic DC showed a decreased capacity to present antigen in vitro, consistent with their reduced major histocompatibility complex class II expression and impaired maturation profile. The accumulation of immature DC may contribute to disease and may reflect an adaptive advantage for the virus by favoring its replication and preventing the generation of fully functional antiviral responses. PMID- 14557660 TI - Evidence against an essential role of COPII-mediated cargo transport to the endoplasmic reticulum-Golgi intermediate compartment in the formation of the primary membrane of vaccinia virus. AB - Vaccinia virus assembles two distinct lipoprotein membranes. The primary membrane contains nonglycosylated proteins, appears as crescents in the cytoplasm, and delimits immature and mature intracellular virions. The secondary or wrapping membrane contains glycoproteins, is derived from virus-modified trans-Golgi or endosomal cisternae, forms a loose coat around some intracellular mature virions, and becomes the envelope of extracellular virions. Although the mode of formation of the wrapping membrane is partially understood, we know less about the primary membrane. Recent reports posit that the primary membrane originates from the endoplasmic reticulum-Golgi intermediate compartment (ERGIC). According to this model, viral primary membrane proteins are cotranslationally inserted into the ER and accumulate in the ERGIC. To test the ERGIC model, we employed Sar1(H79G), a dominant negative form of the Sar1 protein, which is an essential component of coatomer protein II (COPII)-mediated cargo transport from the ER to the ERGIC and other post-ER compartments. Overexpression of Sar1(H79G) by transfection or by a novel recombinant vaccinia virus with an inducible Sar1(H79G) gene resulted in retention of ERGIC 53 in the ER but did not interfere with localization of viral primary membrane proteins in factory regions or with formation of viral crescent membranes and infectious intracellular mature virions. Wrapping of intracellular mature virions and formation of extracellular virions did not occur, however, because some proteins that are essential for the secondary membrane were retained in the ER as a consequence of Sar1(H79G) overexpression. Our data argue against an essential role of COPII-mediated cargo transport and the ERGIC in the formation of the viral primary membrane. Instead, viral membranes may be derived directly from the ER or by a novel mechanism. PMID- 14557661 TI - Metaphase chromosome tethering is necessary for the DNA synthesis and maintenance of oriP plasmids but is insufficient for transcription activation by Epstein-Barr nuclear antigen 1. AB - Epstein-Barr Virus (EBV) infects resting B cells, within which it establishes latency as a stable, circular episome with only two EBV components, the cis element oriP and the latently expressed protein EBNA1. It is believed that EBNA1's ability to tether oriP episomes to metaphase chromosomes is required for its stable replication. We created fusions between the DNA-binding domain (DBD) of EBNA1 and the cellular chromatin-binding proteins HMGA1a and HMG1 to determine the minimal requirements for stable maintenance of an oriP-based episome. These two proteins differ in that HMGA1a can associate with metaphase chromosomes but HMG1 cannot. Interestingly, coinciding with metaphase chromosome association, HMGA1a-DBD but not HMG1-DBD supported both the transient replication and stable maintenance of oriP plasmids, with efficiencies quantitatively similar to that of EBNA1. However, HMGA1a-DBD activated transcription from EBNA1-dependent episomal reporter to only 20% of the level of EBNA1. Furthermore, EBNA1 but not HMGA1a-DBD activated transcription from a chromosomally integrated EBNA1-dependent transcription reporter. This indicates that EBNA1 possesses functional domains that support transcription activation independent of its ability to tether episomal oriP plasmids to cellular chromosomes. We provide evidence that metaphase chromosome tethering is a fundamental requirement for maintenance of an oriP plasmid but is insufficient for EBNA1 to activate transcription. PMID- 14557662 TI - Active borna disease virus polymerase complex requires a distinct nucleoprotein to-phosphoprotein ratio but no viral X protein. AB - Analysis of the composition and regulation of the Borna disease virus (BDV) polymerase complex has so far been limited by the lack of a functional assay. To establish such an assay on the basis of an artificial minigenome, we constructed expression vectors encoding either nucleoprotein (N), phosphoprotein (P), X protein, or polymerase (L) of BDV under the control of the chicken beta-actin promoter. A Flag-tagged version of L colocalized with virus-encoded N and P in characteristic nuclear dots of BDV-infected cells and increased viral N-protein levels in persistently infected Vero cells. Vector-driven expression of L, N, and P in BSR-T7 cells together with a negative-sense BDV minigenome carrying a chloramphenicol acetyltransferase (CAT) reporter gene resulted in efficient synthesis of CAT protein. Induction of CAT protein synthesis strongly depended on a 10- to 30-fold molar excess of the N-encoding plasmid over the P-encoding plasmid. Cotransfection of even small amounts of plasmid encoding the viral X protein reduced CAT synthesis to background levels. Thus, the N-to-P stoichiometry seems to play a central role in the regulation of the BDV polymerase complex. Our data further suggest a negative regulatory function for the X protein of BDV. PMID- 14557663 TI - Norwalk virus nonstructural protein p48 forms a complex with the SNARE regulator VAP-A and prevents cell surface expression of vesicular stomatitis virus G protein. AB - Norwalk virus (NV), a reference strain of human calicivirus in the Norovirus genus of the family Caliciviridae, contains a positive-strand RNA genome with three open reading frames. ORF1 encodes a 1,789-amino-acid polyprotein that is processed into nonstructural proteins that include an NTPase, VPg, protease, and RNA-dependent RNA polymerase. The N-terminal protein p48 of ORF1 shows no significant sequence similarity to viral or cellular proteins, and its function in the human calicivirus replication cycle is not known. The lack of sequence similarity to any protein in the public databases suggested that p48 may have a unique function in the NV replication cycle or, alternatively, may perform a characterized function in replication by a unique mechanism. In this report, it is shown that p48 displays a vesicular localization pattern in transfected cells when fused to the fluorescent reporter EYFP. A predicted transmembrane domain at the C terminus of p48 was not necessary for the observed localization pattern, but this domain was sufficient to redirect localization of EYFP to a fluorescent pattern consistent with the Golgi apparatus. A yeast two-hybrid screen identified the SNARE regulator vesicle-associated membrane protein-associated protein A (VAP A) as a binding partner of p48. Biochemical assays confirmed that p48 and VAP-A interact and form a stable complex in mammalian cells. Furthermore, expression of the vesicular stomatitis virus G glcyoprotein on the cell surface was inhibited when cells coexpressed p48, suggesting that p48 disrupts intracellular protein trafficking. PMID- 14557664 TI - Astrovirus induces diarrhea in the absence of inflammation and cell death. AB - Astroviruses are a leading cause of infantile viral gastroenteritis worldwide. Very little is known about the mechanisms of astrovirus-induced diarrhea. One reason for this is the lack of a small-animal model. Recently, we isolated a novel strain of astrovirus (TAstV-2) from turkeys with the emerging infectious disease poult enteritis mortality syndrome. In the present studies, we demonstrate that TAstV-2 causes growth depression, decreased thymus size, and enteric infection in infected turkeys. Infectious TAstV-2 can be recovered from multiple tissues, including the blood, suggesting that there is a viremic stage during infection. In spite of the severe diarrhea, histopathologic changes in the intestine were mild and there was a surprising lack of inflammation. This may be due to the increased activation of the potent immunosuppressive cytokine transforming growth factor beta during astrovirus infection. These studies suggest that the turkey will be a useful small-animal model with which to study astrovirus pathogenesis and immunity. PMID- 14557665 TI - Adenovirus E1B 55-kilodalton oncoprotein binds to Daxx and eliminates enhancement of p53-dependent transcription by Daxx. AB - The adenovirus E1B 55-kDa protein impairs the p53 pathway and enhances transformation, although the underlying mechanisms remain to be defined. We found that Daxx binds to the E1B 55-kDa protein in a yeast two-hybrid screen. The two proteins interact through their C termini. Mutation of three potential phosphorylation sites (S489/490 and T494 to alanine) within the E1B 55-kDa protein did not affect its interaction with Daxx, although such mutations were previously shown to inhibit E1B's ability to repress p53-dependent transcription and to enhance transformation. In addition to their coimmunoprecipitation in 293 extracts, purified Daxx interacted with the E1B 55-kDa protein in vitro, indicating their direct interaction. In 293 cells, Daxx colocalized with the E1B 55-kDa protein within discrete nuclear dots, where p53 was also found. Such structures were distinct from PML (promyelocytic leukemia protein) bodies, and it appeared that Daxx was displaced from PML bodies. Thus, the Daxx concentration was diminished in dots with a prominent presence of PML and vice versa. Indeed, PML overexpression led to dramatic redistribution of Daxx from p53-E1B 55-kDa protein complexes to PML bodies. Additionally, expression of the E1B 55-kDa protein in Saos2 osteosarcoma cells reduced the number of PML bodies. Our data suggest that E1B and PML compete for available Daxx in the cell. Surprisingly, Daxx significantly augmented p53-mediated transcription and the E1B 55-kDa protein eliminated this effect. Thus, it is likely that the E1B 55-kDa protein sequesters Daxx and p53 in specific nuclear locations, where p53 cannot activate transcription. One consequence of the Daxx-E1B interaction might be an alteration of normal interactions of Daxx, PML, and p53, which may contribute to cell transformation. PMID- 14557666 TI - Dengue virus induces novel changes in gene expression of human umbilical vein endothelial cells. AB - Endothelial cells are permissive to dengue virus (DV) infection in vitro, although their importance as targets of DV infection in vivo remains a subject of debate. To analyze the virus-host interaction, we studied the effect of DV infection on gene expression in human umbilical vein endothelial cells (HUVECs) by using differential display reverse transcription-PCR (DD-RTPCR), quantitative RT-PCR, and Affymetrix oligonucleotide microarrays. DD identified eight differentially expressed cDNAs, including inhibitor of apoptosis-1, 2'-5' oligoadenylate synthetase (OAS), a 2'-5' OAS-like (OASL) gene, galectin-9, myxovirus protein A (MxA), regulator of G-protein signaling, endothelial and smooth muscle cell-derived neuropilin-like protein, and phospholipid scramblase 1. Microarray analysis of 22,000 human genes confirmed these findings and identified an additional 269 genes that were induced and 126 that were repressed more than fourfold after DV infection. Broad functional responses that were activated included the stress, defense, immune, cell adhesion, wounding, inflammatory, and antiviral pathways. These changes in gene expression were seen after infection of HUVECs with either laboratory-adapted virus or with virus isolated directly from plasma of DV-infected patients. Tumor necrosis factor alpha, OASL, and MxA and h-IAP1 genes were induced within the first 8 to 12 h after infection, suggesting a direct effect of DV infection. These global analyses of DV effects on cellular gene expression identify potentially novel mechanisms involved in dengue disease manifestations such as hemostatic disturbance. PMID- 14557667 TI - The hepatitis B virus polymerase mutation rtV173L is selected during lamivudine therapy and enhances viral replication in vitro. AB - Therapy of chronic hepatitis B virus (HBV) infection with the polymerase inhibitor lamivudine frequently is associated with the emergence of viral resistance. Genotypic changes in the YMDD motif (reverse transcriptase [rt] mutations rtM204V/I) conferred resistance to lamivudine as well as reducing the in vitro replication efficiency of HBV. A second mutation, rtL180M, was previously reported to partially restore replication fitness as well as to augment drug resistance in vitro. Here we report the functional characterization of a third polymerase mutation (rtV173L) associated with resistance to lamivudine and famciclovir. rtV173L was observed at baseline in 9 to 22% of patients who entered clinical trials of adefovir dipivoxil for the treatment of lamivudine resistant HBV. In these patients, rtV173L was invariably found as a third mutation in conjunction with rtL180M and rtM204V. In vitro analyses indicated that rtV173L did not alter the sensitivity of wild-type or lamivudine-resistant HBV to lamivudine, penciclovir, or adefovir but instead enhanced viral replication efficiency. A molecular model of HBV polymerase indicated that residue rtV173 is located beneath the template strand of HBV nucleic acid near the active site of the reverse transcriptase. Substitution of leucine for valine at this residue may enhance polymerization either by repositioning the template strand of nucleic acid or by affecting other residues involved in the polymerization reaction. Together, these results suggest that rtV173L is a compensatory mutation that is selected in lamivudine-resistant patients due to an enhanced replication phenotype. PMID- 14557668 TI - Hendra virus V protein inhibits interferon signaling by preventing STAT1 and STAT2 nuclear accumulation. AB - The V protein of the recently emerged paramyxovirus, Nipah virus, has been shown to inhibit interferon (IFN) signal transduction through cytoplasmic sequestration of cellular STAT1 and STAT2 in high-molecular-weight complexes. Here we demonstrate that the closely related Hendra virus V protein also inhibits cellular responses to IFN through binding and cytoplasmic sequestration of both STAT1 and STAT2, but not STAT3. These findings demonstrate a V protein-mediated IFN signal evasion mechanism that is a general property of the known Henipavirus species. PMID- 14557669 TI - Binding of transmissible gastroenteritis coronavirus to brush border membrane sialoglycoproteins. AB - Transmissible gastroenteritis coronavirus (TGEV) is a porcine pathogen causing enteric infections that are lethal for suckling piglets. The enterotropism of TGEV is connected with the sialic acid binding activity of the viral surface protein S. Here we show that, among porcine intestinal brush border membrane proteins, TGEV recognizes a mucin-type glycoprotein designated MGP in a sialic acid-dependent fashion. Virus binding assays with cryosections of the small intestine from a suckling piglet revealed the binding of TGEV to mucin-producing goblet cells. A nonenteropathogenic mutant virus that lacked a sialic acid binding activity was unable to bind to MGP and to attach to goblet cells. Our results suggest a role of MGP in the enteropathogenicity of TGEV. PMID- 14557670 TI - Multiple viral determinants mediate myopathogenicity in coxsackievirus B1-induced chronic inflammatory myopathy. AB - Mice infected with myopathic coxsackievirus B1 Tucson (CVB1(T)) develop chronic inflammatory myopathy (CIM) consisting of hind limb weakness and inflammation. Amyopathic virus variants are infectious but attenuated for CIM. In this report, viral clones, chimeras, and sequencing were used to identify viral determinants of CIM. Chimeras identified several regions involved in CIM and localized a weakness determinant to nucleotides 2493 to 3200 of VP1. Sequencing of multiple clones and viruses identified five candidate determinants that were strictly conserved in myopathic viruses with one located in the 5' untranslated region (UTR), three in the VP1 capsid, and one in the 3C protease. Taken together, these studies implicate Tyr-87 and/or Val-136 as candidate determinants of weakness. They also indicate that there are at least two determinants of inflammation and one additional determinant of weakness encoded by myopathic CVB1(T). PMID- 14557671 TI - Retrotransposition and cell-to-cell transfer of foamy viruses. AB - A remarkable feature of the prototype foamy virus (PFV) replication pathway has been reported to consist of the ability to retrotranspose intracellularly with high efficiency (M. Heinkelein, T. Pietschmann, G. Jarmy, M. Dressler, H. Imrich, J. Thurow, D. Lindemann, M. Bock, A. Moebes, J. Roy, O. Herchenroder, and A. Rethwilm, EMBO J. 19:3436-3345, 2000). PFV intracellular retrotransposition (IRT) was reported to be enhanced by coexpression of fusion-defective envelope protein. To investigate the possibility of cell-to-cell transfer of PFV genomes, which could mimic IRT, we performed cocultivation experiments with cells transfected with an IRT-competent and marker gene-expressing PFV vector together with cells expressing a different marker and measured cells positive for both markers. The findings corroborated the initial report on IRT of Env-deficient PFV. Furthermore, they indicated that viral cores that have incorporated fusion deficient Env can be transferred from cell to cell in a cell type-specific manor. One possible explanation consists of a minor alternative cleavage site in Env that can be used to expose the fusion peptide of the Env transmembrane protein, which appears to be required for virus uptake. PMID- 14557672 TI - Induction of protective immunity against malaria by priming-boosting immunization with recombinant cold-adapted influenza and modified vaccinia Ankara viruses expressing a CD8+-T-cell epitope derived from the circumsporozoite protein of Plasmodium yoelii. AB - We immunized mice with an attenuated (cold-adapted) influenza virus followed by an attenuated vaccinia virus (modified vaccinia virus Ankara), both expressing a CD8(+)-T-cell epitope derived from malaria sporozoites. This vaccination regimen elicited high levels of protection against malaria. This is the first time that the vaccine efficacy of a recombinant cold-adapted influenza virus vector expressing a foreign antigen has been evaluated. PMID- 14557674 TI - Regulation of the human leukemia inhibitory factor gene by ETS transcription factors. AB - OBJECTIVES: Leukemia inhibitory factor (LIF) is a pleiotropic cytokine mainly produced by activated T lymphocytes. We previously demonstrated that human Jurkat T lymphoma cells represent a valid model of LIF gene expression. This study was designed to identify regions critical for LIF promoter activation in Jurkat cells. METHODS: Luciferase constructs under the control of different portions of the human LIF promoter were transfected into Jurkat cells, and promoter activity was determined by luminometry. Similar experiments were performed with constructs bearing mutations in the putative ETS binding regions in the LIF promoter. RT PCR, Western blot and gelshift experiments were performed to study expression and DNA binding of ETS factors in lymphoid cells. RESULTS: With the exception of the shortest construct not including the putative ETS binding sites, all wildtype LIF promoter constructs were strongly inducible by phorbol ester/ionomycin. In contrast, the mutant constructs were significantly less inducible. Cotransfection of the wild-type constructs with ETS expression vectors resulted in significant enhancement of promoter activity. ets-1 and ets-2 mRNA and protein were shown to be expressed in Jurkat cells. Gelshift experiments revealed that proteins present in nuclear extracts from Jurkat cells specifically bind to both artificial ETS consensus sites and ETS binding sites present in the LIF promoter. CONCLUSIONS: We conclude that binding of ETS transcription factors to the ETS binding sites in the human LIF promoter is critical for its inducibility in response to T cell activators. ETS transcription factors thus play an important functional role within the endocrine-immune network. PMID- 14557673 TI - Anterior pituitary cells express pattern recognition receptors for fungal glucans: implications for neuroendocrine immune involvement in response to fungal infections. AB - OBJECTIVES: Hormones and cytokines are known to act as regulatory messengers between the neuroendocrine and immune systems. The innate immune system identifies infectious agents by means of pattern-recognition receptors. These receptors recognize pathogen-specific macromolecules called pathogen-associated molecular patterns. Fungal cell wall glucans nonspecifically stimulate various aspects of innate immunity via interaction with membrane receptors on immune competent cells. Glucans are also released into the systemic circulation of patients with fungal infections. Recent evidence confirms the existence of glucan specific receptors on cells outside the immune system. We hypothesized that glucans may directly interact with pituitary cells as an early signaling event in fungal infections. METHODS: We characterized the receptor-mediated interaction of glucan derived from Candida albicans with pituitary cells using surface plasmon resonance. Prolactin levels were assayed by commercial ELISA. TLR2, TLR4 and CD14 mRNA levels were assessed by RT-PCR. RESULTS: A single glucan-specific binding site was identified on rodent somatomammotroph (K(D) = 3.9 microM) and human folliculostellate cell (K(D) = 3.6 microM) membranes. Coincubation of glucan with somatomammotroph cells for 72 h significantly (p < 0.01) increased prolactin accumulation by 56-62% over that observed in cells treated with media alone. Glucan also increased TLR4 and CD14 gene expression in human folliculostellate cells. CONCLUSIONS: Pituitary cells directly recognize and respond to fungal cell wall glucans resulting in stimulation of pituitary cell TLR4 and CD14 gene expression. In addition, glucan stimulates secretion of prolactin, a hormone that plays an important role in the response to infection. PMID- 14557675 TI - Regulation of allergic lung inflammation in rats: interaction between estradiol and corticosterone. AB - OBJECTIVE: One third of asthmatic women report a decreased expiratory peak flow during menses. Since asthma is characterized by lung inflammation and bronchopulmonary hyperresponsiveness, we investigated the role played by estradiol in allergic lung inflammation. METHODS: Cell migration to the lungs of allergic female rats subjected to oophorectomy (OVx) was compared to that in their sham-operated (sham) control counterparts. Seven days after OVx or sham operation, the rats were sensitized intraperitoneally with ovalbumin (OA, 1 mg/kg) suspended in aluminum hydroxide (day 0). At day 7, a subcutaneous booster of OA was performed and an aerosolized OA challenge was carried out at day 14. One day later (day 15), the rats were killed and cell counts were performed in bronchoalveolar lavages (BAL), in peripheral blood and in bone marrow lavages. RESULTS: After the antigen challenge, OVx rats showed a significant decrease in cell migration to the lung as compared to sham-operated rats. Differential analyses of BAL revealed a reduced number of eosinophils, mononuclear cells and neutrophils. In contrast, in bone marrow as well as in the peripheral blood the numbers of eosinophils, mononuclear cells and neutrophils were increased relative to sham controls. Mast cell numbers were similar in both groups. The estradiol receptor antagonist tamoxifen decreased the allergic lung inflammation in intact rats down to levels similar to those found in untreated OVx rats. In contrast, 17beta-estradiol replacement in OVx rats reestablished the allergic lung inflammation, as observed by an elevated number of eosinophils, mononuclear cells and neutrophils recovered in BAL. Similarly, an elevated number of inflammatory cells were quantified in BAL from allergic OVx rats when corticosterone effects were blocked with metyrapone or RU-486. CONCLUSION: Our results suggest that estradiol has proinflammatory actions on the allergic lung response, and these actions seem to be mediated, at least in part, by endogenous glucocorticoids. PMID- 14557676 TI - Hypothalamic response to experimental allergic encephalomyelitis: role of substance P. AB - Adjuvant-induced arthritis (AA) is thought to be a model for experimental chronic stress that has as main features decreased adrenocorticotropin hormone (ACTH) plasma levels and a rise in median eminence content of arginine vasopressin (AVP) due to the activity of substance P. In experimental allergic encephalomyelitis (EAE), another chronic stress model, the role of substance P action is not clear. In this paper we tried to clarify the role of substance P in Lewis rats, which are susceptible to this disease. EAE was induced using myelin basic protein plus complete Freund's adjuvant injected into the hind limbs. One day later injections of an antagonist to substance P (RP 67580), saline, and substance P were administered daily for 12-14 days through a stainless steel cannula into the lateral ventricle of the brain, and then the rats were killed. The rats were divided into groups of controls, sham, diseased controls (no intracerebroventricular injections) and EAE (injected intracerebroventricularly). Plasma was used for the quantification of ACTH and corticosterone but not AVP which was assayed in hypothalamic median eminence extracts. In noninjected diseased rats the plasma levels of ACTH and corticosterone were significantly higher than in noninjected control rats, whereas the AVP concentrations in the median eminence were unchanged. The substance P antagonist did not affect the levels of these hormones in plasma or the median eminence. Substance P decreased the plasma levels of ACTH and corticosterone but did not increase the median eminence content of vasopressin. Administration of the antagonist 30 min before an equivalent dose of substance P increased the plasma levels of the two hormones, but did not change the content of AVP. Based on the lack of response to the antagonist RP 67580 we suggest that the substance P has different roles in EAE and AA at least in the later stages of EAE (after 11 days of immunization). PMID- 14557677 TI - Granulocyte macrophage colony-stimulating factor regulates activity of the nervous system. AB - OBJECTIVES: The peripheral administration of granulocyte macrophage colony stimulating factor (GM-CSF), widely used for the treatment of cytopenias, is often associated with neurological effects [Lieschke et al., N Engl J Med 1992;327:28-34]. This cytokine has recently been reported to affect neurotransmitter metabolism in the nervous system [Bianchi, Neuroreport 1997;8:3587-3590]. To further investigate the neuromodulatory effect of GM-CSF we studied the influence of GM-CSF on the efferent electric activity in the splenic nerve and the integral neuronal activity in medullary gigantocellular reticular formation (MGRF) in rats. METHODS: Anaesthetized (sodium thiopental 70 mg/kg, i.p.) Wistar rats were injected subcutaneously with 1 microg/kg of hr GM-CSF. Efferent electric activity in the splenic nerve and integral electric activity in MGRF were analyzed. The effectiveness of the applied dose of GM-CSF was verified by determining the elevation of the white blood cell count in peripheral blood 60 min after injection. RESULTS: We found that GM-CSF increases efferent electric activity in the splenic nerve and decreases that of MGRF as is evident by the frequency of electric discharges. The latency of both effects was 5-15 min. CONCLUSIONS: This data support the view that GM-CSF exerts a neuromodulatory effect and may provide a new link of neuroimmune communication. PMID- 14557678 TI - Inhibition of decrease in natural killer cell activity in repeatedly restraint stressed mice by a biological response modifier derived from cultured mycelia of the basidiomycete Tricholoma matsutake. AB - OBJECTIVE: To develop a method to cope with stress-induced reduction in immunocompetence, we evaluated the immunomodulatory activities of a biological response modifier derived from the mycelia of the basidiomycete Tricholoma matsutake (CM6271) in mice under repeated restraint stress. METHODS: C57BL/6 mice were inserted, one per tube, into 50-ml polypropylene tubes into which more than 30 ventilation holes had been drilled, and were restrained everyday for 20 days in this fashion for set periods of time. Natural killer (NK) cell activity and NK1.1-positive cell counts in the spleen, ACTH and corticosterone levels in the blood were determined. CM6271 was orally administered daily during the restraint stress period. RESULTS: (1) When the mice were restrained in a confined space for 6 h per day for 20 days, the NK cell activity and the NK1.1-positive cell counts in the spleen significantly decreased after day 5 with an increase in the blood ACTH and corticosterone levels. (2) Oral administration of CM6271 during the restraint stress period significantly prevented the stress-induced decrease in NK cell activity. The effect was dependent on the timing, duration, and doses administered. (3) CM6271 did not significantly affect the splenic NK1.1-positive cell counts or the levels of blood ACTH and corticosterone in restraint-stressed mice. CONCLUSION: The above findings suggest that CM6271 inhibits the restraint stress-induced decrease of NK cell activity in a timing of administration and dose-dependent manner. PMID- 14557679 TI - Cohabitation with a sick cage mate: consequences on behavior and on ehrlich tumor growth. AB - The present study analyzed the effects of cohabitation for 11 days with a sick cage mate on behavior and Ehrlich tumor growth in mice. Pairs of female mice were divided into one control and one experimental group. One mouse of each control pair was kept undisturbed and called 'healthy companion' (HC). One animal of each experimental pair of mice was inoculated (i.p.) with 5 x 10(6) Ehrlich tumor cells, and the other, the object of this study, was called 'sick companion' (SC). The SC mice presented: (1) increased activity in an open field, (2) increased number of entries and of movements within the plus-maze open arms, (3) similar levels of plus-maze closed-arm exploration, (4) a decrease in the exploratory activity in a hole board, (5) a decrease in the number of white but not red blood cells, and (6) similar corticosterone serum levels. Eleven days after cohabitation with a conspecific, HC and SC mice were injected with 5 x10(6) Ehrlich tumor cells. Results showed that SC animals presented decreased resistance to the ascitic form of the Ehrlich tumor. The observed data provide experimental evidence that psychosocial stress induced by cohabitation with a sick cage mate changed at the same time some behavioral and physiological parameters, and decreased resistance to Ehrlich tumor. These data are discussed in the light of a possible neuroimmune system interaction. PMID- 14557680 TI - The sympathetic control of blood supply is different in the spleen and lymph nodes. AB - OBJECTIVES: The noradrenergic innervation of lymphoid organs controls several immune cell functions and local blood perfusion. Considering that cell and antigen uptake depend on the blood supply to lymphoid organs, the hypothesis was tested that feedback signals from activated immune cells control sympathetic vasomotor activity. METHODS: We determined the blood flow in spleen and mesenteric lymph nodes (mLN) of Wistar Kyoto rats during immune stimulation with endotoxin (LPS; 10 microg/kg) and following disruption of the noradrenergic transmission. RESULTS: Our data indicate that (a) the splenic noradrenaline content, which reflects the density of the sympathetic innervation, is 5 times higher in the spleen than in other peripheral organs and the spleen receives stronger tonic sympathetic input than mLN; (b) immune stimulation with LPS causes a 4-fold increase in the IL-1beta production in the spleen, but only 2-fold in mLN; (c) IL-1beta causes an inhibition of the sympathetic vasoconstrictor tonus in the spleen, but has no significant effect on the noradrenergic vascular tonus in mLN, and (d) in mLN, the local hyperemia induced by LPS is attenuated by the degranulation of vesicular stores of histamine and serotonin, indicating that these monoamines participate in the vasodilator effect of LPS in mLN. CONCLUSIONS: The present experiments, taken together with our previous studies, indicate that the control of blood supply to the spleen and mLN involves different mechanisms. While blood perfusion in the spleen depends on the inhibition of the noradrenergic vasoconstriction by endogenously produced IL 1beta, other vasoactive mediators such as serotonin and histamine play a role in the control of mLN perfusion. PMID- 14557681 TI - Neuroepidemiology increases impact factor. PMID- 14557682 TI - Impaired motor speed, visuospatial episodic memory and verbal fluency characterize cognition in long-term stroke survivors: the Tromso Study. AB - The cognitive function after stroke is examined in acute and subacute phase, but poorly characterized in long-term stroke survivors. This paper discusses cognitive function among long-term stroke survivors, with matched stroke-free subjects, based on a population survey. General cognition, verbal, executive and visuospatial function, memory, attention, and motor speed were tested as well as motor function in upper extremities. Stroke survivors and controls were most effectively discriminated by means of motor speed, followed by visuospatial episodic memory and verbal fluency. This pattern of cognitive disturbances may be a consequence of cerebral lesions in frontal subcortical areas, and is different from Alzheimer's disease. PMID- 14557683 TI - Risk factors of stroke mortality: a 40-year follow-up of the Corfu cohort from the Seven-Countries Study. AB - The present work evaluates several risk factors for stroke mortality based on a 40-year follow-up of the Corfu cohort from the Seven-Countries Study. The population studied in this analysis consisted of 529 rural men (49.7 +/- 5.7 years old) enrolled at 1961. The death rate at the end of follow-up was 87.1% (i.e. 461 deaths out of 529 participants). Of 529 cardiovascular-disease-free men at entry, 74 (14%) died because of hemorrhagic or thrombotic stroke. Age (HR=1.11/year, p<0.001), pulse pressure (HR=1.16/5 mm Hg, p<0.001), total serum cholesterol levels (HR=0.81/10 mg/dl, p<0.01) and presence of physical activity (HR=0.59, p<0.05), showed a statistically significant association with fatal stroke events. In conclusion, age, pulse pressure levels, physical activity (protective) and total serum cholesterol levels (protective) were significantly related with 40-year stroke mortality. PMID- 14557685 TI - Viral meningitis-associated hospitalizations in the United States, 1988-1999. AB - We used the National Hospital Discharge Survey and the Nationwide Inpatient Sample of the Health Care Cost and Utilization Project to estimate disease burden associated with viral meningitis hospitalizations in the United States. During 1988-1999, viral meningitis accounted for an estimated 434,000 hospitalizations (annual average, 36,000; average annual hospitalization rate, 14/100,000), and 2.1 million hospital days (annual average, 175,000). The estimated mean charge for viral meningitis-associated hospitalization during 1993-1997 varied between USD 6,562 and 8,313, resulting in annual estimated hospitalization costs between USD 234 and 310 million and a total estimated cost of nearly USD 1.3 billion for the 5-year period. In summary, viral meningitis remains an important cause of morbidity and financial burden and merits efforts to improve diagnostic, treatment, and prevention options. PMID- 14557684 TI - Alcohol and hemorrhagic stroke in Santiago, Chile. A case-control study. AB - BACKGROUND AND PURPOSE: Hemorrhagic stroke (HS) is a major cause of disability and death worldwide. There is a dearth of information on HS from geographically defined populations in Latin America. In this study we assessed the importance of alcohol consumption as a risk factor for HS in Chile. METHODS: Case-control study in Santiago, Chile, of 140 consecutive patients with CT-confirmed HS, matched by sex and age with 140 hospital controls. Alcohol consumption was measured in grams (ethanol) per week, using a questionnaire administered to the patients or caregivers or both. We defined four categories of alcohol consumption: nondrinkers (0.0 g/week), light (0.1-115 g/week), moderate (116-402.5 g/week) and heavy drinkers (>402.5 g/week). Other variables measured included diabetes mellitus (DM), cigarette smoking, arterial hypertension, liver disease and chronic use of nonsteroidal anti-inflammatory drugs (NSAID). Statistical analysis was performed with STATA 6.0 software. RESULTS: A total of 280 subjects with a mean age of 65.5 years were studied over a 3-year period, 122 men (43.5%) and 158 women (56.5%). Alcohol intake was 394.1 g/week among cases and 174.5 g/week in controls (p=0.01). The following odds ratios (OR) with 95% confidence intervals (CI) were found: hypertension 4.89 (2.86-10.3) and chronic use of NSAID 3.44 (2.15-12.9). Using conditional logistic regression analysis high alcohol intake was found to have a statistically significant OR of 4.47 (CI 1.14-17.2). CONCLUSIONS: In Chile, a high alcohol intake (>402.5 g/week) increased more than 4 times the risk of HS and remained a significant risk factor for HS after controlling for hypertension, cigarette smoking, liver disease, blood cholesterol levels, and chronic use of NSAID. The risk was higher in younger patients (<65 years of age). PMID- 14557686 TI - Occupational exposure to metals and solvents and the risk of motor neuron disease. A case-control study. AB - Previous studies based on recalled occupational histories have implicated metal and solvent exposure in the etiology of motor neuron disease (MND). We have used death certificates held in pension fund archives and linked unbiased historical occupational records to investigate the effects of occupational exposure to metals or solvents on the risk of death from MND in 22 cases and 206 controls from 22,526 past employees of a major UK engineering company. We found no evidence of increased risk of death from MND in individuals who had worked with either metals (adjusted odds ratio=0.88, 95% CI 0.35-2.22) or solvents (odds ratio=1.12, 95% CI 0.45-2.78), and no relation between disease risk and either duration or intensity of exposure. We conclude that metal and solvent exposures are unlikely to be involved in the etiology of MND. PMID- 14557687 TI - Parkinson's disease and CYP2D6 polymorphism in Asian populations: A meta analysis. AB - Polymorphism of CYP2D6 and its relationship with the development of Parkinson's disease (PD) has been controversial. The distribution of the B-mutation of CYP2D6, a mutation that results in the absence of a functional protein, differs by ethnicity and accounts for less than 1% of the 'poor metabolizer' phenotype in Asians. Thus, a meta-analysis was conducted to determine if polymorphism, other than the B-mutation, within the CYP2D6 gene confers a greater susceptibility to PD outcome among Asian populations. Eleven studies were identified, two of which were excluded due to unavailability in the English language or availability of the same original data in more detail in another publication. None of the studies showed a statistically significant association between CYP2D6 polymorphism and PD (p<0.05). The overall odds ratio was 0.84 (95% confidence interval 0.66-1.08). We conclude that among Asian populations, there is no convincing evidence of an association between CYP2D6 polymorphism and the risk of developing PD. PMID- 14557688 TI - Multiple sclerosis in Northern Sardinia, Italy: a methodological approach for genetic epidemiological studies. AB - The etiopathogenesis of multiple sclerosis (MS) remains unclear. However, genetic factors are believed to be important in disease susceptibility. A methodological approach is presented for a population-based study aimed at investigating MS familial incidence and patterns of familial clustering in Northern Sardinia, Italy, with a reported MS prevalence of 150/100,000 population. Patients with MS since 1965 to the present and known to the MS Register for the province of Sassari will be asked to provide genealogical, comorbid and demographic information. Statistical analyses of the familial risk for MS will depend on the completeness of MS 'age at onset' data for affected individuals and age at the time family history is obtained (or age at death) for unaffected family members. PMID- 14557689 TI - Chemotherapy for high-grade glioma. PMID- 14557690 TI - Exercise-induced human coronary collateral function: quantitative assessment during acute coronary occlusions. AB - In 50 patients undergoing percutaneous transluminal coronary angioplasty because of chronic angina pectoris, a collateral flow index (CFI) was determined at the start and the end of two 1-min coronary occlusions, randomly accompanied by a resting state or a 3-min dynamic handgrip exercise (DHE). CFI expressing collateral flow relative to normal antegrade flow was determined by simultaneous coronary occlusive pressure, mean aortic pressure and central venous pressure measurements. When comparing CFI without and with DHE at the start as well as at the end of balloon occlusions, a significant increase was observed with DHE (overall p < 0.0001); start: 0.18 +/- 0.12 vs. 0.22 +/- 0.13, respectively (p = 0.01); end of occlusion: 0.21 +/- 0.14 vs. 0.25 +/- 0.14, respectively (p = 0.007). PMID- 14557691 TI - A syndrome of transient left ventricular apical wall motion abnormality in the absence of coronary disease: a perspective from the United States. AB - BACKGROUND: The syndrome of chest pain associated with characteristic anterior electrocardiographic changes, moderate increases in cardiac enzymes, and a reversible apical wall motion abnormality in the absence of coronary artery disease has been documented in Japan, but has received relatively little attention in other countries. METHODS: The clinical and echocardiographic data of 12 patients (11 women, mean age 64 +/-14 years) who presented with chest symptoms, electrocardiographic (ECG) changes indicative of an acute anteroapical myocardial infarction, abnormal cardiac enzyme levels and echocardiography showing an apical wall motion abnormality were collected. Coronary angiography was performed in 10 patients. A follow-up echocardiogram was obtained within 2 weeks of the initial diagnosis in most cases. RESULTS: An identifiable, precipitating ("trigger") event could be identified in all 12 individuals. Respiratory distress was present in 7, the death of a relative in 3, in 4 a surgical or medical procedure had been performed, and in 1 a panic disorder was diagnosed. The echocardiograms showed a characteristic wall motion pattern of significant apical dysfunction. All of the patients who underwent coronary arteriography had noncritical coronary artery disease. Follow-up echocardiography showed normalization of the LV dysfunction in all instances. CONCLUSION: We identified a syndrome of chest pain, dyspnea, ECG and enzyme changes mimicking acute myocardial infarction, similar to the "Takotsubo" syndrome described in Japan. It is likely that the widespread use of echocardiography, coupled with increased recognition of this syndrome, will result in this diagnosis being made more commonly. PMID- 14557692 TI - The influence of bradycardia and normofrequent ventricular pacing on myocardial blood flow and energy demand in rats. AB - The purpose of the present study was to determine the effect of beta-blockade induced bradycardia and normofrequent ventricular pacing (VP) on regional myocardial (MBF) and cerebral blood flow (CBF) as well as on myocardial oxygen consumption (MVO(2)) using colored microspheres. The MBF at sinus rhythm (SR) was 4.60 +/- 1.27 ml/g/min and decreased to 3.80 +/- 0.74 ml/g/min (p < 0.05) during beta-blockade. The MBF was 5.10 +/- 1.88 ml/g/min during VP with beta-blockade. The endo-/epicardial MBF ratio during VP was significantly reduced. MVO(2) at beta-blockade was significantly lower (0.21 +/- 0.06 ml/g x min(-1), p < 0.05) than at SR (0.47 +/- 0.15 ml/g x min(-1)) and during VP with beta-blockade (0.54 +/- 0.14 ml/g x min(-1)). RESULTS: (i) MBF and MVO(2) are reduced with decreasing mean arterial pressure (MAP) during beta-blockade-induced bradycardia, but MBF and MVO(2) are significantly increased during normofrequent VP with beta-blockade despite a further reduction in MAP. (ii) The MBF ratio is constant during beta blockade but is significantly lower during normofrequent VP with beta-blockade. (iii) CBF remained constant during all experimental conditions. CONCLUSION: Despite beta-blockade, the subendocardium is jeopardized during normofrequent VP by ischemia. PMID- 14557693 TI - Regional heterogeneity in gene expression profiles: a transcript analysis in human and rat heart. AB - BACKGROUND: Investigations involving biopsies of human cardiac tissue often assume that myocardial samples from a specific location are representative of the entire heart. HYPOTHESIS: There are significant regional differences in gene expression in the heart. METHODS: We used two models. In the first model, seven whole human hearts were cut in 1-cm slices from apex to base and 11 distinct regions were sampled. Full thickness left ventricular tissue was further subdivided equally into an inner, outer, and middle region. In the second model, hearts were removed from adult Sprague-Dawley rats and were divided into 4 regions. Using species-specific quantitative reverse transcriptase-polymerase chain reaction, we measured transcript levels of myosin heavy chain beta (MHC beta), glucose transporter 1 (GLUT 1), and atrial natriuretic factor (ANF) in tissue samples from both models. RESULTS: In human heart, there were significant differences in transcript levels between regions. The following patterns could be recognized among the seven hearts. ANF expression was highest in the subendocardial region. MHC-beta and GLUT 1 transcript levels were higher in the right ventricle than the left ventricle. As expected, ANF transcript levels were highest in the atria, where MHC-beta and GLUT 1 expression was low. Analogous to the human studies, MHC-beta and GLUT 1 transcript levels were low in rat atria as compared to ventricles. In rat heart, MHC-beta expression was higher in the left ventricle than the right ventricle while GLUT 1 expression was not significantly different between ventricles. CONCLUSION: Despite the large variability in transcript levels among different regions in human hearts, certain patterns in gene expression emerged suggesting that different anatomical regions of the heart also differ in respect to gene expression. PMID- 14557694 TI - Smooth muscle cell outgrowth from coronary atherectomy specimens in vitro is associated with less time to restenosis and expression of a key Transcription factor KLF5/BTEB2. AB - Atherectomy specimens offer an opportunity to study the biology of coronary artery lesions. We cultured smooth muscle cells (SMCs) from specimens obtained from 24 patients with coronary restenosis after angioplasty to study the relationship between activity of SMCs (in vitro outgrowth) and the time course of restenosis. We also examined expression of a Kruppel-like zinc-finger transcription factor 5 (KLF; also known as BTEB2 and IKLF), which is markedly induced in activated SMCs, in the same specimens. SMC outgrowth was observed in 9 of 24 specimens (37.5%). Restenosis occurred sooner (p < 0.01) in patients whose specimens showed outgrowth compared to those whose specimens showed no outgrowth. Immunostaining for KLF5 was more common in specimens with outgrowth (89 vs. 20%, p < 0.01). These data suggest that the number of activated SMCs in lesions may determine in vitro outgrowth and also affect the time to restenosis. PMID- 14557695 TI - T-wave alternans in patients with right ventricular tachycardia. AB - Microvolt T-wave alternans has been proposed as a new risk marker for ventricular arrhythmias. However, the clinical significance of T-wave alternans in patients with ventricular tachycardia (VT) originating from the right ventricle has been unknown. The study population consisted of 20 patients with arrhythmogenic right ventricular cardiomyopathy (ARVC) or idiopathic VT. T-wave alternans was measured during bicycle exercise testing using the CH 2000 system. Of the 7 patients with ARVC, 6 (86%) were positive for T-wave alternans. On the other hand, only 1 (8%) of 13 patients with idiopathic VT originating from the right-ventricular outflow tract was positive for T-wave alternans. PMID- 14557696 TI - Dobutamine stress echocardiography in hypertrophic cardiomyopathy. AB - AIMS: Myocardial ischemia in the absence of coronary artery disease is common in patients with hypertrophic cardiomyopathy (HCM). Dobutamine stress echocardiography (DSE) induces left ventricular (LV) new wall motion abnormalities (NWMA) in some patients with HCM. We evaluated the effects of dobutamine on LV performance and hemodynamics in HCM. METHODS AND RESULTS: Eighteen patients with non-obstructive HCM underwent DSE. Dobutamine was administered at dosages of 5, 10, 20, 30 and 40 microg/kg/min with increments at intervals of 3 min. Seven patients developed NWMA, whereas the other 11 did not. During DSE, heart rate increased significantly more in NWMA patients,whereas LV outflow tract gradient (OTG) increased significantly and similarly in both groups. At peak dobutamine dose, NWMA patients had a significant increase in LV end-systolic diameter and volume and a significant decrease in LV fractional shortening and ejection fraction. Posterior wall thickening increased significantly, whereas septal thickening did not increase throughout DSE in both groups. CONCLUSIONS: In a subgroup of patients with HCM, DSE induces NWMA, associated with a greater increase in heart rate, irrespective of LVOTG. NWMA induce a depression of global LV systolic performance. The septum shows no contractile reserve, regardless of NWMA. These phenomena may be the result of induction of ischemia and/or impaired LV systolic function due to fast heart rate. PMID- 14557697 TI - Circadian variation of QT dispersion determined by twelve-lead Holter electrocardiography. PMID- 14557698 TI - "Apoptoses en masse" form Condorelli's prestatic myocardial infarction. PMID- 14557699 TI - Atrial fibrillation in hypertension: an established association with several unresolved issues. PMID- 14557700 TI - Sudden cardiac death in a hunger striker. PMID- 14557701 TI - Home care technology and telehealth--the future is here! PMID- 14557702 TI - Seven performance-accelerating technologies that will shape the future of home care. PMID- 14557703 TI - What is data decision support and why should I care? PMID- 14557704 TI - Telehealth opportunities for home care patients. PMID- 14557705 TI - Assessing technology needs for the elderly: finding the perfect match for home. PMID- 14557706 TI - Connecting for compliance: improving satisfaction and care coordination by extending the home care record to the physician. PMID- 14557707 TI - The Cardiac Connection program: home care that doesn't miss a beat. PMID- 14557708 TI - The American Telemedicine Association and home care. PMID- 14557709 TI - The impact of technology on the "older" nurse. PMID- 14557710 TI - A picture speaks louder than words...but a digital camcorder tells the whole story. PMID- 14557711 TI - Interactive personal technology education program decreases adverse medication events. PMID- 14557712 TI - Telehealth and risk management in home health. PMID- 14557713 TI - First home telehealth clinical guidelines developed by the American Telemedicine Association. PMID- 14557714 TI - OASIS Web- and CD-based training now available. PMID- 14557715 TI - Home care's challenge: move the information...not the patient. PMID- 14557716 TI - Effects of stimulus salience and methamphetamine on choice reaction time in the rat: central tendency versus distribution skew. AB - Stimulants decrease reaction time in humans as well as laboratory rats. This effect is seen as a decrease in average reaction time or a shift in the distribution peak towards shorter reaction times. However, response-time distributions are typically skewed, exhibiting a positive tail. Our goal for this project was to develop a method of analyzing reaction-time distributions in the rat which will allow us to study systematically measures of central tendency and distribution skew. This analysis subdivided reaction time into initiation time and movement time, and also subdivided the response time distributions into distribution mode and distribution skew. Rats were trained on a two-choice visual reaction-time task. We then evaluated the effects of stimulus salience and methamphetamine (METH) treatment (vehicle, 0.25, 0.5, 1.0, and 1.5 mg/kg) on measures of distribution mode and skew. Stimulus salience decreased initiation time mode, initiation time skew and movement time skew, but had no effect on movement time mode. METH had a greater effect on skew for the initiation time distribution and a greater effect on mode for the movement time distribution. This analysis will serve as a useful method of determining whether initiation time and movement time, as well as distribution mode and distribution skew, represent different behavioral processes in the rat. PMID- 14557718 TI - Effects of the delta-opioid receptor agonist SNC80 on learning relative to its antidepressant-like effects in rats. AB - Delta-opioid receptor agonists produce decreases in immobility in the forced swim test, suggesting that these compounds have antidepressant-like activity. There is also the possibility that these compounds decrease immobility in the forced swim test by disrupting learning processes that occur during the swim, or with successive swim exposures, thus falsely identifying them as having "antidepressant" potential. This study investigated the effects of the delta opioid receptor agonist, SNC80, on responding in a repeated-acquisition procedure and in the forced swim test in rats, and the effects were compared directly to those of scopolamine, a compound known to disrupt memory and learning. SNC80 disrupted acquisition of a response sequence (learning) and produced a significant antidepressant-like effect in the forced swim test. Scopolamine, however, produced larger decrements in learning without producing behavioral changes consistent with an antidepressant-like profile of action. These results suggest that SNC80 produces antidepressant-like activity through a mechanism independent of its disruptive effects on learning. PMID- 14557717 TI - Characterization of scratching responses in rats following centrally administered morphine or bombesin. AB - The aim of this study was to characterize scratching behavior elicited by central administration of morphine or bombesin in rats, and to determine the role of opioid receptors in scratching induced by both pruritogenic agents. Central administration included intracisternal (i.c.), intrathecal (i.t.), and intracerebroventricular (i.c.v.) routes. Scratching events made with hind paws were counted by observers blinded to treatment conditions. Intracisternal morphine (0.01-0.1 microg) produced dose-dependent increases in scratching; the maximum response to i.c. morphine 0.1 microg was approximately 500 scratches within a 1-hour period. Neither i.t. nor i.c.v. morphine significantly increased scratching. Bombesin (0.01-0.32 microg) elicited robust scratching following i.c. administration. The maximum response to i.c. bombesin 0.32 microg was approximately 4000 scratches within a 1-hour period. Both i.t. and i.c.v. bombesin produced profound scratching at similar doses. Antagonist studies confirmed that mu-opioid receptors selectively mediate i.c. morphine-induced scratching. However, selective mu-, kappa-, and delta-opioid antagonists did not attenuate i.c. bombesin-induced scratching. These results demonstrate that morphine and bombesin elicit scratching through different receptor mechanisms, at different central sites, and to different degrees. PMID- 14557719 TI - Systemic and intracerebroventricular administration of sodium barbital induced a place preference in rats. AB - We have shown previously that 15 mg/kg pentobarbital induces a conditioned place preference (CPP), but it is unsuitable for intracranial administration. Since the long-acting barbiturate, sodium barbital, is soluble at a neutral pH, we tested whether it would induce a CPP when administered centrally. Furthermore, because barbital has a long duration of action, and because we obtained a significant CPP to systemically administered barbital using 30-minute conditioning trials, we tested whether longer conditioning trials would produce a more robust CPP. Using a three-compartment apparatus and an unbiased procedure, we found that systemic administration of barbital induced a significant CPP at 8 and 24 mg/kg, but not 2.7 or 72 mg/kg (i.p.). When rats were conditioned to 24 mg/kg barbital for conditioning trials of (1/2), 1, 3, or 6 hours, only the 30-min conditioning trial produced a CPP. Finally, 240 and 480 microg intracerebroventricular (ICV) barbital induced a significant CPP, but 60 or 120 microg did not. These findings suggest that: (1) like pentobarbital, barbital has reinforcing properties measured in the CPP test; (2) the CPP is impaired, rather than enhanced, by increasing the duration of drug-context pairing; and (3) the reinforcing effects of barbiturates are centrally mediated. PMID- 14557720 TI - Facilitation and disruption of memory for the passive avoidance task in the day old chick using dopamine D1 receptor compounds. AB - This series of studies provides a behavioural account of dopamine D1-receptor dependent facilitation and disruption of memory for the single-trial passive avoidance task in the day-old chick. The D1 antagonist, SCH23390, induced memory disruption in a dose-dependent manner from 60 min after training with a strong (100% methyl anthranilate) aversant experience. The D1 agonist, SKF38393, was found to facilitate memory in chicks given a weak (20% vol/vol methyl anthranilate) training experience. The D2 antagonist, sulpiride, and the D2 agonist, quinpirole, showed no memory effects. The research indicates an important role for dopamine D1-dependent mechanisms in memory formation in the chick. PMID- 14557722 TI - Effects of fluoxetine on social behaviour and plasma corticosteroid levels in female Mongolian gerbils. AB - Gerbils are a highly social species and extremely sensitive to social manipulations. In this laboratory, separating male/female pairs has been found to produce significant effects on these animal's subsequent social behaviour. The present studies were conducted in order to examine the effects of a short period of individual housing in females of this species, as this may also be predicted to produce alterations in social responding. It was found that 21 days' individual housing induced a marked reduction in social behaviour directed towards an untreated male placed in the same arena. This was indicated by a highly significant increase in immobile-in-contact, a behaviour that involves females freezing while, and only while, they are being socially investigated. This represents the declining of an invitation to socially interact and so high levels of immobile-in-contact indicate low levels of social motivation. There was also an increase in evading, upon another animal's approach, and a decrease in social investigation of other animals. The effects of 15 days of fluoxetine were found to be highly dependent on housing condition. In individually housed females, 10 mg/kg increased their social investigation of other animals and markedly reduced the duration of immobile-in-contact. Twenty mg/kg also reduced levels of immobile-in-contact and increased the frequency of active approaches towards other animals. Fluoxetine therefore acts to increase social motivation in individually housed animals. By direct contrast, in group-housed female gerbils, fluoxetine had no effects on social behaviour and produced clear indications of sedation. While housing condition had no influence on levels of corticosterone, fluoxetine produced dose-related increases in corticosteroid levels in both group and individually housed animals. These findings show that: (1) a short period of individual housing induces a significant reduction in these animals' motivation towards social behaviour; (2) the effects of fluoxetine on behaviour are greatly influenced by housing condition--prosocial effects are seen in individually housed animals but only sedative effects are seen in animals maintained in groups; and (3) while housing condition has no effects on levels of corticosterone, fluoxetine dose-dependently stimulates corticosteroid release. It can be concluded that the effects of fluoxetine on gerbil behaviour are independent of its stimulatory influence on HPA axis functioning, and that the prosocial effects of this selective serotonin reuptake inhibitor (SSRI) can only be seen in animals with a pre-existing social deficit. PMID- 14557721 TI - Repeated administration of phencyclidine, amphetamine and MK-801 selectively impairs spatial learning in mice: a possible model of psychotomimetic drug induced cognitive deficits. AB - Cognitive deficits are a key feature of schizophrenia. N-Methyl-D-aspartate (NMDA) receptor antagonists and amphetamine are known to induce psychotic behaviors and cognitive deficits in animals and humans, often affecting visuo spatial abilities. Phencyclidine (PCP), MK-801 and amphetamine (AMPH) have been used in pharmacological animal models of schizophrenia, but none of these models has focused so far on spatial learning after repeated administration of the drugs. The objective of this study was to test whether repeated administration of PCP, AMPH or MK-801 influenced the performance of mice in a non-associative spatial learning test. CD-1 male mice were given i.p. daily injections of either saline, PCP (5.0, 10.0 mg/kg), AMPH (2.5, 5 mg/kg) or MK-801 (0.3, 0.6 mg/kg), for 5 days. On day 6 all mice were tested in an open field containing five different objects. After three sessions of habituation, each animal's reactivity to object displacement and object substitution was assessed. No significant differences among treatment groups were observed in object exploration or locomotion during the habituation phase. Five days of repeated PCP, AMPH or MK 801 administration selectively and differentially impaired the ability of mice to discriminate a spatial change, while leaving intact the ability to react to a non spatial change. These data suggest that neurobiological adaptations to drug regimens known to induce psychotic behaviors and alterations in locomotor activity or stereotypies can also alter spatial learning, as assessed in this test, thus indicating that these regimens could also mimic some of the cognitive deficits observed in schizophrenia. PMID- 14557723 TI - Similar effects of cocaine and immobilization stress on the levels of heat-shock proteins and stress-activated protein kinases in the rat hippocampus, and on swimming behaviors: the contribution of dopamine and benzodiazepine receptors. AB - Cocaine (COC) has been reported to cause effects similar to physiological stressors in the brain neuroendocrinal system, including heat-shock protein (HSP) expression, although these effects have not been elucidated in detail. In the present study, we examined the effects of repeated (4 days) treatments with cocaine hydrochloride (35 mg/kg, i.p.) and 10 min immobilization stress (IM) on the distribution of HSP (HSP27, HSP60, HSP70, HSC70) and stress-activated protein kinase (SAPK) (SAPKalpha, SAPKbeta, SAPKgamma) immunoreactive nerve cells (positive cells) in the rat hippocampus. The swimming behaviors of the rats in the forced swimming test were also examined. In both COC and IM groups, an early enhancement (5 h time point) of hippocampal HSP (HSP27, HSP60, HSP70, HSC70) and SAPK (SAPKbeta, SAPKgamma) positive cells was observed, whereas a recovery (SAPKs) or attenuation (HSP60 and HSC70) was observed at the 24 h time point. In both groups, a depression of the swimming behaviors (attenuation in the activity counts and time until immobility) below the control level was observed at the 5 h point, but a recovery was observed at the 24 h time point. At the 48 h time point, all parameters returned to the control level. These alterations in the levels of HSPs and SAPKs, and the swimming behaviors were similar to those observed in the stress (IM) group, and were characteristic in that all of these alterations were attenuated by the benzodiazepine inverse agonist, Ro 15-4513 (5 mg/kg, i.p.), and the dopamine D1 receptor antagonist, SCH23390 (0.5 mg/kg, i.p.), which was not observed in the groups treated with another stressor-like drug (bicuculline). PMID- 14557724 TI - Context-dependent catalepsy intensification is due to classical conditioning and sensitization. AB - Haloperidol-induced catalepsy represents a model of neuroleptic-induced Parkinsonism. Daily administration of haloperidol, followed by testing for catalepsy on a bar and grid, results in a day-to-day increase in catalepsy that is completely context dependent, resulting in a strong placebo effect and in a failure of expression after a change in context. The aim of this study was to analyse the associative learning process that underlies context dependency. Catalepsy intensification was induced by a daily threshold dose of 0.25 mg/kg haloperidol. Extinction training and retesting under haloperidol revealed that sensitization was composed of two components: a context-conditioning component, which can be extinguished, and a context-dependent sensitization component, which cannot be extinguished. Context dependency of catalepsy thus follows precisely the same rules as context dependency of psychostimulant-induced sensitization. Catalepsy sensitization is therefore due to conditioning and sensitization. PMID- 14557725 TI - Disruption of a drug-induced choice behavior by UV light. AB - A considerable body of evidence suggests that UV light disrupts ligand binding in vitro. In vivo, UV light effects have been reported to disrupt simple behaviors such as spontaneous locomotor activity. However, there are no reports of UV light blocking a more complex drug-altered behavior. We now report that: (1) cocaine dose-relatedly reversed planarians' usual selection of dark over light (from 19.4+/-4.4% to 73.3+/-6.7%) (this effect was not due to an increase in locomotor activity, since cocaine only minimally increases locomotor activity at the highest dose tested); and (2) the cocaine-induced alteration of behavioral choice was attenuated significantly (P<0.05) by UV light (366 nm and 254 nm). The results demonstrate alteration of a relatively complex choice behavior by UV light. PMID- 14557726 TI - Tobacco update: scientific advances, clinical perspectives. PMID- 14557727 TI - Taking on tobacco: the need for a comprehensive approach. PMID- 14557728 TI - The genetics of smoking related behavior: a brief review. AB - Smoking behavior is influenced by both genetic and environmental factors. Many years of twin and adoption studies have demonstrated that heritability is at least 50% for both smoking initiation and smoking persistence. Furthermore, the extent to which genetic and environmental factors contribute to smoking behavior in men is significantly different from that in women. It has been reported that polymorphisms of candidate genes, such as cytochrome P450, dopamine receptor and transporter, and serotonin transporter genes, are associated with smoking behavior. However, many of these reports have not yet received independent confirmation. Based on the results from genome-wide linkage and association analyses on different subject populations, 15 loci located on 8 chromosomes were reported to harbor susceptibility genes for nicotine and other substances of abuse. It is expected that a newly completed human genome sequence, as well as advances in genomic technologies such as genotyping and array analysis, will greatly enhance progress toward the identification of genes associated with smoking behavior. PMID- 14557729 TI - Cigarette smoke-induced airway inflammation as sampled by the expired breath condensate. AB - Airways exposed to smoke respond with inflammatory processes. The airway inflammation generally present in smokers causes persistent cough and phlegm production, reactive airway disease, and tissue infiltration by inflammatory cells. Although the short-term response may be protective, long-term pathological consequences include swelling of the airway epithelium, mucus hypersecretion, and increased airway reactivity characteristic of chronic bronchitis and chronic obstructive lung disease and the tissue destruction characteristic of emphysema. The natural history of these diseases is poorly understood, because human airway tissue is available for study only at autopsy, from surgical specimens, or from procedures such as bronchoscopy or thoracotomy. A noninvasive method of monitoring the inflammation is by analyzing expired breath condensate, which contains a diluted sample of airway surface liquid. The study of expired breath condensate may offer a more practical approach to sampling airway chemistry and make it possible to study the detailed inflammatory response to airborne particulates. PMID- 14557730 TI - Oral changes associated with tobacco use. AB - Tobacco is a delivery system for the addictive agent nicotine. The dental profession is encouraged to perform oral examinations that focus on oral cancer detection, but other oral changes occur with tobacco use. The oral mucosa is composed of stratified squamous epithelium and masticatory/keratinized (hard palate, dorsum of the tongue, and keratinized gingival) and lining mucosa (floor of the mouth, ventrolateral surface of the tongue, soft palate complex, labial vestibule, and buccal mucosa). Tobacco use affects the surface epithelium, resulting in changes in the appearance of the tissues. The changes may range from an increase in pigmentation to thickening of the epithelium (white lesion). Tobacco use can also irritate the minor salivary glands on the hard palate and directly increase a person's risk for periodontal disease and oral cancer. This article will review some of the more common oral lesions that are associated with tobacco use-smoker's melanosis, nicotinic stomatitis, periodontal disease, smokeless tobacco keratosis, gingival recession/tooth abrasion, black hairy tongue, and oral cancer. PMID- 14557731 TI - Treating nicotine dependence. AB - This article reviews the biology of nicotine addiction and the most effective treatments for nicotine dependence. Nicotine elevates dopamine levels in the brain, which is associated with drug reinforcement and dependence. Nicotine abuse leads to nicotine dependence. The most effective pharmacological adjuncts for treating nicotine dependence are nicotine replacement therapy and bupropion SR. When these are combined with behavioral counseling, the best treatment outcomes are observed. PMID- 14557732 TI - Smoking and depression in Chinese Americans. AB - BACKGROUND: A close link between smoking and depression has been documented by research primarily based on U.S. white populations. This study examined the association between depressive symptoms and smoking behaviors in Chinese American smokers. METHODS: Analyses were based on baseline data collected from a convenience sample of 199 Chinese smokers who resided in northern California. The sample included 20.6% women, 97% immigrants, with mean age of 40.8 years and mean cigarettes/day of 8.9. RESULTS: Compared with population-based studies of Chinese Americans, the Chinese smokers in the current study reported higher depressive symptoms assessed by the Center for Epidemiologic Studies-Depression Scale [M, 20.4; 95% confidence interval (CI), 18.8-22.2], higher lifetime prevalence rates of major depressive disorders (30.3%; 95% CI, 24.0-37.2%) and dysthymia (11.6%; 95% CI, 7.5-16.9%). Multiple regression analysis suggested female gender, unemployment, major depression or dysthymia within the past year, previous experience with nicotine withdrawal syndrome, and high temptation to smoke under negative affect situations are associated with a higher level of depressive symptoms. CONCLUSIONS: The level of depressive symptoms among Chinese American smokers is comparable with that observed in other US populations reported. In the current sample, elevated depressive symptoms were more prominent among women or those who were unemployed, smokers who reported significant nicotine withdrawal at previous quit attempts, and high temptation to smoke when experiencing negative emotions. Findings support further examination of the role of depression in smoking among Chinese Americans and underscore the importance of addressing depressive symptoms when treating tobacco use in Chinese smokers. PMID- 14557733 TI - Smoking cessation in women with cardiac risk. AB - This article summarizes what is known about smoking among women, in particular the consequences of smoking on women's cardiac health. Smoking is recognized as a major independent cause of coronary heart disease, a condition that ranks first among the causes of death for middle-aged and older women. Women who stop smoking greatly reduce their risk of heart disease and other smoking-related illnesses. Research since the 1980 report of the US Surgeon General has been inconsistent in finding differences by gender in smoking cessation, and few approaches have been devised specifically for women. Several factors are of unique concern among women and potential barriers to smoking cessation, including weight concerns, negative affect regulation, and hormonal influences. More research is needed to determine whether these factors, when targeted specifically, improve cessation success rates among women. PMID- 14557734 TI - It ain't over till it's over: the case for offering relapse-prevention interventions to former smokers. AB - Most people who attempt to quit tobacco smoking eventually relapse. Although treatment strategies have been developed to prevent smoking relapse, they tend to be available only to the small proportion of smokers who enroll in intensive smoking cessation treatments. It is argued that freestanding relapse-prevention interventions could be offered to persons who recently ceased smoking, whether they used a formal treatment program or quit on their own. A line of research is described demonstrating that a series of relapse-prevention booklets mailed to recent quitters significantly reduces smoking relapse. Moreover, the intervention seems to be highly cost-effective. If disseminated widely, such an approach has the potential to make a significant public health impact. PMID- 14557735 TI - Quitlines in North America: evidence base and applications. AB - Quitlines provide a model for the translation of research findings to public health application. Quitlines are currently in operation in more than half of US states, in Canada, and in multiple countries globally. Overall, when implemented correctly, quitlines have been shown to be efficacious and effective. Multiple quitline models are in use, but there is no evidence on the relative effectiveness of one over the other. Differences have been demonstrated for the efficacy of quitlines for specific applications, with the strongest evidence base for application as a primary intervention or as follow-up for hospitalized patients and particularly for cardiac patients. The evidence base for both reactive and proactive services is reviewed, and future directions to continue to advance the field are discussed. PMID- 14557737 TI - Characteristics of African American smokers: a brief review. AB - The Public Health Service Guideline, Treating Tobacco Use and Dependence, contains a wealth of information regarding current treatment practices. Unfortunately, guidance for tailoring standardized treatments to meet the needs of specific ethnic subgroups is limited. This article highlights features unique to African American smokers, a necessary precursor to developing intervention programs that hold promise for improving outcomes. PMID- 14557738 TI - Smoking cessation or reduction in pregnancy treatment methods: a meta-evaluation of the impact of dissemination. AB - Active and passive exposure to tobacco smoke during pregnancy and infancy is the most serious and preventable cause of adverse maternal, fetal, and infant outcomes in the United States. The multiple risks of tobacco exposure to mothers and infants are definitive, and the clinical and economic benefits of cessation have been documented. This article provides a synopsis of the state of the science and art in this specialized area and reviews the evidence for validity of patient assessment methods and the effectiveness of smoking cessation/reduction treatments for pregnant women. A synthesis of 4 topics is presented: (1) the validity of patient reports of smoking status and recent trends during pregnancy and postpartum; (2) definition of "Best Practice" smoking cessation methods for pregnant women; (3) the effect of dissemination of effective clinical practice methods among the 800,000+ pregnant US smokers each year; and (4) the evidence for the cost-benefit of improved maternal and infant outcomes from cessation. PMID- 14557736 TI - What have we learned from 20 years of research on smokeless tobacco cessation? AB - Although smoking cessation continues to get most of the attention from both researchers and practitioners, treatment for smokeless tobacco (SLT) dependence gets little consideration. The reasons are varied, but essentially smoking is more prevalent, has greater public health implications, and has been the subject of clean air laws and environmental issues that have focused the attention on a burned tobacco, thereby leaving SLT with scant public health attention. This is unfortunate, because the sales of SLT products, especially moist snuff, have increased consistently over the past decade; regular use of these products can result in oral cancer, other negative effects on oral health, and nicotine dependence or addiction. This article will review some unique issues of SLT use that affect cessation and the empirical research on interventions. PMID- 14557739 TI - Serious mental illness and tobacco addiction: a model program to address this common but neglected issue. AB - Tobacco addiction among persons with serious mental illness (SMI) has been largely ignored. About 75 to 85% of persons with schizophrenia, bipolar disorder, and other SMI use tobacco; most will either die and/or have reduced quality of life because of tobacco-caused medical diseases. Tobacco addiction is the most common co-occurring disorder for the SMI population. A dramatic reduction in tobacco use in the general population has occurred during the past 40 years; however, there has been almost no reduction for smokers with SMI. The University of Medicine and Dentistry of New Jersey program targets smokers with SMI and provides outreach services, clinical treatment and research, and consultation to other community-based mental health treatment agencies in New Jersey. Clinical and research evidence supports motivation-based treatment, blending mental health and addiction treatment approaches, and integrating tobacco dependence treatment within mental health settings. The unique barriers and clinical issues for this population are described. PMID- 14557740 TI - The female smoker: from addiction to recovery. AB - Millions of American girls and women have been drawn to smoking by an industry that has been clearly and systematically targeting women of all ages and life circumstances. Big tobacco's well-timed marketing strategies skillfully link cigarette use to typical female values: independence, self-reliance, weight control, stress management, social progress and popularity, personal attractiveness, autonomy, self-fulfillment, youth, happiness, personal success, health, and active, vigorous, and strenuous lifestyles. Biologically speaking, women are especially vulnerable to the legion of health problems of tobacco use. Smoking is a critical hazard for women in their reproductive years, particularly when they are pregnant. The US Public Health Service 2000 Clinical Practice Guideline provides helpful guidance and sound general recommendations for the treatment of women of all ages for tobacco use and dependence. Women and girls who smoke represent diverse subgroups of the population with unique issues and needs. The 2001 Surgeon General's Report on Women and Smoking stresses the importance of multistrategy programs for treating female smokers. This approach includes antitobacco media campaigns, increases in tobacco prices, promotion of nonsmoking in public places, curbs on tobacco advertising and promotion, enforcement of legislation to reduce youth access to tobacco products, and effective tobacco use treatment programs. PMID- 14557741 TI - A brief review of adolescents and tobacco: what we know and don't know. AB - Smoking among teenagers is a major public health problem, and the purpose of this article is to review the literature concerning etiological factors of teen smoking, effects of prevention programs on reducing rates of teenage smoking, and the efficacy of smoking cessation treatment. Peer, family, and intrapersonal factors all play a role in the continuum of initiation to regular use. Prevention programs focusing on social norms governing teenage smoking are particularly effective in reducing onset of smoking behavior. Finally, behavioral cessation programs have been shown to be effective in reducing teenage smoking. Future directions concerning prevention and cessation research are discussed. PMID- 14557742 TI - Smoking cessation research in primary care treatment centers: the SCRIPT-MS project. AB - Despite wide distribution of an evidence-based clinical practice guideline, the provision of treatment for tobacco use has been weak. The primary care setting is an ideal environment in which to implement the tobacco clinical practice guideline. It has been suggested that implementation of the guideline may be enhanced by adapting guideline recommendations into a stepped-care plus treatment matching model; however, this model has yet to be tested. This article describes an ongoing investigation designed to evaluate (1) the feasibility of implementing this treatment model in a primary care environment, (2) efforts on identifying reasonable assessment methods, (3) primary care providers' use of assessment data, and (4) the relationships between predictors and outcomes under differing treatment "step" conditions in primary care settings. Six primary care clinics are participating in this study comparing usual care, brief clinical intervention, and enhanced clinical intervention conditions. The last of these conditions is hypothesized to produce the best and most cost-effective outcomes. Recruitment is expected to continue until January 2004, being 65% complete as of this writing. Follow-up contacts will continue until April 2005. Results should provide information that will contribute to the ongoing development of primary care-based tobacco intervention approaches. PMID- 14557743 TI - Healthcare provider training in tobacco treatment: building competency. AB - One of the most effective disease prevention and health promotion strategies available is the treatment of tobacco dependence. The Agency for Healthcare Research and Quality (AHRQ) clinical practice guideline for treating tobacco use and dependence provides a number of recommendations for interventions by health care systems and providers, including that treatment be reimbursed, identification of tobacco users be institutionalized, and all patients be offered brief treatment and have more intensive treatment available to them. Unfortunately, these recommended interventions have not been routinely implemented. As part of a comprehensive approach to improving implementation, provider training is needed. Three models for training healthcare providers in brief tobacco treatment intervention (medical education, professional education, and community-based education) are described, a model for training providers in intensive tobacco treatment interventions is presented, and a rationale for a national training and certification program is made. PMID- 14557744 TI - Trends and patterns of tobacco use in the United States. AB - This review summarizes recent trends and current patterns of tobacco use in the United States. Although adult smoking dropped between 1965 and 1990, from 50% to 28% of men and from 35% to 23% of women, the past decade has seen little further progress. In 2000, 25.7% of US men and 21.0% of women were smokers. Adolescent smoking has been declining since the late 1990s, but nearly 30% of high school seniors still smoke. In 2000, 4.4% of US men and 0.3% of women used snuff or chewing tobacco. Although adolescent smokeless tobacco use has declined in recent years, 14.8% of male high school students were current users in 2001. In 2001, 22.1% of male high school students and 8.5% of women students were current cigar smokers. Bidis and kreteks may be gaining popularity among young people, and more than 15% of adolescent smokers use these tobacco products. Despite recent progress, tobacco use remains prevalent in the United States. State and local governments need to invest adequate resources in the full range of tobacco control activities. PMID- 14557745 TI - Public health issues in treating tobacco use. AB - Comprehensive strategies are needed to prevent initiation of tobacco use, to help users quit, and to reduce morbidity, mortality and indirect social costs. Using an agent-host-vector model for disease, all forms of tobacco are the "agent," whether they are manufactured by large companies or are traditional tobacco products. "Host" refers to tobacco users and everyone else because all are in some way adversely affected, including the unborn. The "vector" is the tobacco industry. The public needs truth, tobacco dependent victims require help, and the tobacco industry needs accountability and restraint. PMID- 14557746 TI - Macrophages act as effectors of tissue damage in acute renal allograft rejection. AB - BACKGROUND: Macrophages constitute 38% to 60% of infiltrating cells during acute renal allograft rejection. Their contribution to tissue damage during acute rejection was examined by depleting macrophages in a rat model. METHODS: Lewis rats underwent bilateral nephrectomy and then received a Dark Agouti renal allograft and liposomal-clodronate, control phosphate-buffered saline liposomes, or saline intravenously (n=7 per group) on days 1 and 3 postsurgery. Grafts were harvested on day 5. RESULTS: Liposomal-clodronate treatment resulted in a 70% reduction in blood ED1+ monocytes and 60% reduction in intragraft ED1+ macrophages (both P<0.01). Half of all remaining interstitial ED1+ cells were undergoing apoptosis (terminal deoxynucleotide transferase-mediated dUTP nick-end labeling+/ED1+), and thus functional depletion of more than 75% of macrophages was achieved. Histologic and functional parameters of acute rejection were attenuated: interstitial infiltrate, tubulitis, and glomerulitis (P<0.01); tubular cell apoptosis (P<0.001); tubular cell proliferation (P<0.001); and serum creatinine (P<0.01). Production of inducible nitric oxide synthase by infiltrating cells and urinary nitric oxide excretion was reduced by 90% (P<0.001). In contrast, no reduction in the number of other leukocytes was seen (CD3+, CD4+, CD8+, and natural killer cells). Activation of lymphocytes (CD25+) and production of lymphocyte effector molecules (granzyme B) were unaltered. CONCLUSION: This study demonstrates that macrophages contribute to tissue damage during acute rejection. PMID- 14557747 TI - Specific immune responses against airway epithelial cells in a transgenic mouse trachea transplantation model for obliterative airway disease. AB - BACKGROUND: Immune injury to airway epithelium is suggested to play a central role in the pathogenesis of obliterative bronchiolitis (OB) after clinical lung transplantation. In several studies, a rejection model of murine trachea transplants is used, resulting in obliterative airway disease (OAD) with similarities to human OB. To focus on the role of an immune response specifically against airway epithelium, we transplanted tracheas from transgenic mice expressing human epithelial glycoprotein (hEGP) on epithelial cells. We hypothesized that the immune response against the hEGP-2 antigen would result in OAD in the trachea transplants. METHODS: Tracheas from hEGP-2 transgenic and control nontransgenic FVB/N mice were heterotopically transplanted into FVB/N mice and harvested at week 1, 3, 6, and 9. Anti-hEGP-2 antibodies were determined in the recipient blood. The trachea grafts were analyzed for cellular infiltration, epithelial cell injury, and luminal obliteration. RESULTS: Recipients of transgenic tracheal grafts gradually developed anti-hEGP-2 antibodies. In the transgenic grafts, the submucosa was infiltrated predominantly by CD4+ T cells. Epithelial cells remained present but showed progressive abnormality. The tracheal lumen showed a mild degree of obliteration. All these changes were absent in nontransgenic FVB/N trachea transplants. CONCLUSION: The hEGP-2 antigen on the epithelial cells of transgenic trachea transplants induces specific humoral and cellular immune responses, leading to a mild form of OAD. It provides a suitable model for further investigation of the role of epithelial cells in the development of OAD in animals and OB in human-lung transplantation. PMID- 14557748 TI - Significant prolongation of animal survival by combined therapy of FR167653 and cyclosporine A in rat renal allografts. AB - BACKGROUND: Cyclosporine A (CsA) has improved short-term results in renal transplantation. However, its toxicities have been serious problems in clinical patients undergoing long-term administration. It is necessary to develop a nonnephrotoxic immunosuppressant that allows reducing doses of CsA. The authors tested the effect of a new pyrazolotriazin compound, FR167653 (FR), that inhibits the production of proinflammatory cytokines, using a rat renal acute-rejection model (Wistar-Lewis). METHODS: The authors first examined the adverse reactions of FR in naive rats. The effect of FR monotherapy was evaluated by treating allograft recipients at a dosage of 32 or 64 mg/kg/day intraperitoneally. The synergistic effect of FR and CsA at a minimum dose was examined by treating the recipients daily with 64 mg/kg of FR and 0.1, 0.5, or 1.0 mg/kg of CsA. RESULTS: Long-term administration of FR caused no severe adverse reactions in naive rats at less than 96 mg/kg/day, and FR is a nonnephrotoxic but liver-toxic agent at higher doses. Monotherapy of FR (8.7+/-1.3 days) or CsA (8.7+/-1.7 days) did not influence graft survival (vs. 8.2+/-0.8 in controls, n=10). However, the combined treatment of FR and CsA significantly prolonged animal survival (>50% of animals survived a 42-day follow-up period; P<0.01 vs. all other groups, n=15/group). Immunologic analysis demonstrated the significantly decreased production of major inflammatory mediators and adhesion molecules in the allografts as compared with those of all other groups. CONCLUSIONS: A nonnephrotoxic agent, FR may be a promising candidate for a new combined immunosuppressive regimen that potentially reduces the amount of CsA. PMID- 14557749 TI - NOD/LtSz-Rag1nullPfpnull mice: a new model system with increased levels of human peripheral leukocyte and hematopoietic stem-cell engraftment. AB - BACKGROUND: A critical need exists for effective small-animal models that accept engraftment of human hematopoietic progenitor cells and mature lymphocytes. The purpose of this study was to determine the phenotypic effects of perforin (Pfp) deficiency on nonobese diabetic (NOD)-Rag1null mice and to evaluate the ability of NOD/LtSz-Rag1nullPfpnull recipients to support engraftment with human hematolymphoid cells. METHODS: A new genetic stock of NOD mice doubly homozygous for targeted mutations at the recombination activating gene (Rag)-1 and Pfp genes was developed. NOD/LtSz-Rag1nullPfpnull mice were studied for immunopathologic and hematologic abnormalities. The ability of these mice to support engraftment with human peripheral blood mononuclear cells (PBMC) and umbilical-cord blood hematopoietic progenitor cells was assessed. RESULTS: NOD/LtSz-Rag1nullPfpnull mice lacked mature B cells, T cells, natural killer (NK) cell cytotoxic activity and were devoid of serum immunoglobulin (Ig) throughout a 37-week lifespan. These mice supported heightened engraftment with human PBMC as compared with NOD/LtSz Rag1null controls as evidenced by a 4- to 5-fold increase in percentages of human lymphocytes and a 7- to 13-fold increase in percentages of CD4+ T cells in the peripheral blood and spleen. Total numbers of human CD4+ T cells were increased approximately 20-fold in the spleens of NOD/LtSz-Rag1nullPfpnull mice. These mice also showed approximately 12-fold higher levels of engraftment with human umbilical-cord blood cells compared with NOD/LtSz-Rag1null mice. CONCLUSIONS: NOD/LtSz-Rag1nullPfpnull mice are devoid of mature B cell, T cell, and NK cell cytotoxic activity, engraft at high levels with human PBMC, and hematopoietic progenitor cells and provide a new NK cell-deficient model for human hematolymphoid cell engraftment. PMID- 14557750 TI - Superparamagnetic iron oxide particles transactivator protein-fluorescein isothiocyanate particle labeling for in vivo magnetic resonance imaging detection of cell migration: uptake and durability. AB - Conjugation of dextran-coated superparamagnetic iron oxide (SPIO) particles with transactivator protein (Tat)-peptide and fluorescein isothiocyanate (FITC) allows cells to readily uptake SPIO particles. This makes possible high-resolution, real time imaging of these cells by magnetic resonance imaging (MRI). First, we need to understand how various subpopulations take up and maintain SPIO particles. In this report, we have focused on differences in T cells, B cells, and macrophages with respect to cross-linked (CL)-SPIO Tat-FITC particle uptake over 72 hours. We have found that cells quickly take up the particles and that the bead loss that does occur is not related to cell death or apoptosis. In contrast with reports in the literature, we have observed migration of the Tat-peptide conjugates primarily to the cytoplasm rather than the nucleus. PMID- 14557751 TI - Gemcitabine with cyclosporine or with tacrolimus exerts a synergistic effect and induces tolerance in the rat. AB - BACKGROUND: This study investigated the effect of the antineoplastic agent gemcitabine (dFdC) in combination with cyclosporine (CsA) or with FK506 on acute heart allograft rejection in a rat model. METHODS: Transplantations were performed in the fully allogeneic Lewis-to-Brown Norway strain combination. dFdC, CsA, and FK506 single-drug therapy and combinations of dFdC with CsA and FK506 were administered at various dosages starting on day 1 to prevent and on day 4 to treat acute rejection until day 20. Animals who did not reject their graft were intraperitoneally injected with 108 splenic donor-type lymphocytes. In addition, Lewis and third-party skin grafts were transplanted to these animals. RESULTS: Mean graft survival times under CsA, FK506, and dFdC monotherapy were 18.3/63.7 days (1 mg/5 mg per kg), 41.7 days, and 24.7/38.7 days (100 microg/150 microg per kg), respectively. CsA and FK506 in combination with dFdC prolonged graft survival to more than 100 days (CsA) and more than 95.2 days (FK506). Graft survival after treatment of an ongoing rejection was 21.5/38.3 days for CsA (1 mg/5 mg per kg) and 17.7/59.2 days for dFdC (100 microg/150 microg per kg). The combination of CsA+dFdC prompted indefinite survival of five of six hearts. Lymphocyte inoculation did not induce graft rejection. Notably, none of the Lewis, but all third-party, skin grafts were rejected immediately. Histomorphologic analysis of grafted hearts, however, demonstrated typical features of chronic rejection. CONCLUSIONS: The combination of CsA and FK506 with low-dose dFdC exerts a synergistic effect in the prevention and treatment of acute allograft rejection in this model. Although chronic rejection could not be prevented, strain-specific tolerance was achieved. Therefore, combining standard immunosuppressants with dFdC is a novel, promising strategy for prevention and treatment of acute allograft rejection. PMID- 14557752 TI - Restoration of functional motor units in a rat model of sphincter injury by muscle precursor cell autografts. AB - BACKGROUND: Urinary incontinence is a debilitating condition that affects primarily elderly individuals. One major mechanism results from chronic denervation of the striated urethral sphincter with associated fibrosis. The authors investigated the fate of muscle precursor cells (MPC) injected into a model of striated urethral sphincter injury that reproduces the histopathologic changes of sphincter insufficiency. METHODS: The striated urethral sphincter of older male rats was damaged by electrocoagulation. MPC were isolated from limb myofiber explants, infected with an adenovirus carrying the transgene encoding beta-galactosidase, and injected into the sphincter of the same animal 37 days after injury. Animals were killed 5 and 30 days after injection for assessment of sphincter function and the formation of motor units. RESULTS: Electrocoagulation resulted in an irreversible destruction of both sphincteric myofibers and nerve endings, with a functional incapacity of the damaged sphincter to sustain an increase in bladder pressure; atrophy and fibrosis developed after 1 month. Injection of MPC resulted in the formation of beta-galactosidase-expressing myotubes in the sphincter that persisted beyond 30 days. The regenerated myotubes carried acetylcholine receptors associated with a nerve ending and were thus considered to form anatomic motor units. Urodynamic studies confirmed the restoration of 41% of sphincter function 1 month after MPC injection. CONCLUSIONS: The authors showed that MPC isolated from limb muscles of an older animal can recapitulate a myogenic program when injected into an irreversibly injured sphincter. The maturation of MPC activates nerve regeneration and restores functional motor units. PMID- 14557753 TI - Engraftment and tumor formation after allogeneic in utero transplantation of primate embryonic stem cells. AB - BACKGROUND: To achieve human embryonic stem (ES) cell-based transplantation therapies, allogeneic transplantation models of nonhuman primates would be useful. We have prepared cynomolgus ES cells genetically marked with the green fluorescent protein (GFP). The cells were transplanted into the allogeneic fetus, taking advantage of the fact that the fetus is so immunologically immature as not to induce immune responses to transplanted cells and that fetal tissue compartments are rapidly expanding and thus providing space for the engraftment. METHODS: Cynomolgus ES cells were genetically modified to express the GFP gene using a simian immunodeficiency viral vector or electroporation. These cells were transplanted in utero with ultrasound guidance into the cynomolgus fetus in the abdominal cavity (n=2) or liver (n=2) at the end of the first trimester. Three fetuses were delivered 1 month after transplantation, and the other, 3 months after transplantation. Fetal tissues were examined for transplanted cell progeny by quantitative polymerase chain reaction and in situ polymerase chain reaction of the GFP sequence. RESULTS: A fluorescent tumor, obviously derived from transplanted ES cells, was found in the thoracic cavity at 3 months after transplantation in one fetus. However, transplanted cell progeny were also detected (approximately 1%) without teratomas in multiple fetal tissues. The cells were solitary and indistinguishable from surrounding host cells. CONCLUSIONS: Transplanted cynomolgus ES cells can be engrafted in allogeneic fetuses. The cells will, however, form a tumor if they "leak" into an improper space such as the thoracic cavity. PMID- 14557754 TI - Liver transplantation from Maastricht category 2 non-heart-beating donors. AB - BACKGROUND: The demand for liver transplantation has increasingly exceeded the supply of cadaver donor organs. Non-heart-beating donors (NHBDs) may be an alternative to increase the cadaver donor pool. METHODS: The outcome of 20 liver transplants from Maastricht category 2 NHBDs is compared with 40 liver transplants from heart-beating donors (HBDs). After unsuccessful cardiopulmonary resuscitation (CPR), cardiopulmonary support (CPS) with simultaneous application of chest and abdominal compression (n=6), and cardiopulmonary bypass (CPB; n=14), which was hypothermic (n=7) or normothermic (n=7), were used to preserve the organs from NHBDs. Factors that may influence the outcome of livers from Maastricht category 2 NHBDs were also investigated. RESULTS: With a minimum follow-up of 2 years, actuarial patient and graft survivals with livers from Maastricht category 2 NHBDs were 80% and 55%, respectively. Transplantation of organs from these donors was associated with a significantly higher incidence of primary nonfunction, biliary complications, and more severe initial liver dysfunction compared with livers from HBDs. Graft survival was 83% in livers from NHBDs preserved with CPS and 42% in those maintained with CPB. No graft failed if the duration of warm ischemia did not exceed 130 min with CPR or CPS, and if the period of CPB did not surpass 150 min when this method was used after CPR, regardless if it was hypothermic or normothermic. CONCLUSION: Livers from Maastricht type 2 NHBDs may be used for transplantation if the period of warm ischemia during CPR or CPS does not exceed 130 min. Hypothermic or normothermic CPB after CPR preserves liver viability for an additional 150 min. PMID- 14557755 TI - Value of donor swabs for intra-abdominal infection in simultaneous pancreas kidney transplantation. AB - BACKGROUND: Simultaneous pancreas-kidney transplantation (SPK) has a higher rate of surgical complications compared with other whole organ transplantations. Graft thrombosis and intra-abdominal infections are the most frequent causes for relaparotomy. We evaluated risk factors for abdominal infections after SPK, with emphasis on the value of the routinely taken intraoperative swabs. METHODS: Between June 1994 and December 2000, 177 SPK were performed. Immunosuppression consisted of antithymocyte globulin induction and triple-drug maintenance therapy. Routine swabs were taken from the graft perfusion solutions, from the donors' duodenum, and from the recipients' bladder and jejunum (in case of enteric drainage). RESULTS: A total of 19 (10.7%) of 177 patients underwent 41 relaparotomies as a result of intra-abdominal infections. Positive microbial results from any donor site and positive duodenal swabs were significant risk factors for intra-abdominal infections after SPK (P=0.01, P=0.02). There was a significantly higher incidence of abdominal infections when Candida was found in the donor duodenal swab (P=0.0048). Patient survival was significantly lower in cases with abdominal infection (P=0.02). Survival rates of patients with and without abdominal infection were 89.5% and 97.4% at 1 year and 72.3% and 92.8% at 5 years, respectively. CONCLUSIONS: The results of this study confirm that abdominal infections significantly reduce patient survival and thus jeopardize the success of SPK. Positive donor duodenal swabs have been revealed to be a significant risk factor for a subsequent intra-abdominal infection, especially when Candida was found. PMID- 14557757 TI - Hypophosphatemia after 95 right-lobe living-donor hepatectomies for liver transplantation is not a significant source of morbidity. AB - BACKGROUND: Hypophosphatemia appears to be a universal event after right hepatic lobectomy for live-donor adult liver transplantation according to one report. Because hypophosphatemia appears to contribute to increased postoperative complications, routine hyperalimentation with supratherapeutic levels of phosphorus was advocated. METHODS: From July 2000 to May 2002, we performed 95 right-lobe living-donor hepatectomies for 95 adult liver-transplant recipients, the largest single institutional experience. We reviewed our data that were collected prospectively. RESULTS: We did not find profound hypophosphatemia (<1.0 mg/dL) to be prevalent in our donors. At least six (6.3%) donors did not have postoperative hypophosphatemia. In addition, there appears to be no increased morbidity related to hypophosphatemia when aggressively corrected with intravenous or oral phosphates in our group of donors that underwent right-lobe hepatectomies. CONCLUSIONS: We, therefore, cannot endorse the routine administration of hyperalimentation with supratherapeutic phosphorus because of its potential morbidity. PMID- 14557756 TI - Pharmacodynamics, pharmacokinetics, and safety of multiple doses of FTY720 in stable renal transplant patients: a multicenter, randomized, placebo-controlled, phase I study. AB - BACKGROUND: FTY720, a novel immunomodulator, displays potent immunosuppressive activity in a variety of preclinical transplant models. This study examined the safety, pharmacodynamics, and pharmacokinetics of multiple doses of FTY720 in stable renal transplant patients. METHODS: This randomized, multicenter, double blind, placebo-controlled, phase I study included adults who had been maintained on a regimen of cyclosporine A (CsA) microemulsion and prednisone (or its equivalent) for at least 1 year after renal transplantation. Patients received once-daily doses of 0.125, 0.25, 0.5, 1.0, 2.5, or 5.0 mg FTY720, or placebo for 28 days. After completion of study drug administration, the patients were monitored until day 56 by serial laboratory tests, clinical examinations, and recording of adverse events. The study includes 76 treatment courses (61 FTY720 and 15 placebo), with 65 patients enrolled once and 11 reenrolled. RESULTS: FTY720 doses greater than or equal to 1.0 mg/day produced a significant reduction in peripheral blood lymphocyte count by up to 85%, which reversed within 3 days after discontinuation of study medication. Compared with placebo-treated patients, FTY720 subjects did not show a major increase in adverse events or a change in renal function. Pharmacokinetic measurements revealed that FTY720 displayed linear relations of doses and concentrations over a wide range, but had no effect on CsA exposure. CONCLUSIONS: At doses up to 5.0 mg/day for 28 days, stable renal transplant patients treated with FTY720 in combination with CsA and prednisone displayed a dose-dependent, reversible decline in peripheral blood lymphocytes without an enhanced incidence of collateral toxicities, except possibly bradycardia. PMID- 14557759 TI - Allograft transmission of hepatitis C virus infection from infected donors in cardiac transplantation. AB - BACKGROUND: The frequency and outcome of hepatitis C virus (HCV) infection in recipients of hearts from HCV-infected donors remains poorly characterized. METHODS: Between 1991 and 1999, 10 anti-HCV-negative patients received hearts from donors who were anti-HCV and HCV RNA-positive. Each recipient was tested for anti-HCV and HCV RNA and serially evaluated for liver dysfunction. Recipient records were reviewed for cumulative steroid boluses in the first posttransplant year and other components of the immune suppression regimen. We analyzed recipient outcome in relation to the virologic status of the donor, including the level of HCV RNA and genotype and the type of antirejection therapy. RESULTS: All 10 recipients became HCV RNA positive. Donor-recipient pairs expressed identical genotypes in each instance. Six of nine evaluable recipients developed biochemical evidence of hepatitis. Recipients with genotype 1 (1a, 1b) accounted for five of the six cases, and all patients with genotype 1 developed hepatitis. Severe liver injury occurred in two patients. Two deaths occurred, both of which were genotype 1 patients who had been given multiple boluses of corticosteroids in the first posttransplant year. No definite relationship between viral load in the donor and recipient outcome was found. CONCLUSION: Transmission of HCV infection from cardiac donors who are viremic at the time of organ donation occurs with high frequency and can cause severe hepatitis. Hearts from infected patients should probably be restricted to those recipients who already have evidence for hepatitis C or are in need of emergent transplantation. PMID- 14557758 TI - Long-term acceptance of rat cardiac allografts on the basis of adenovirus mediated CD40Ig plus CTLA4Ig gene therapies. AB - BACKGROUND: We have previously demonstrated that blockade of either CD80/86-CD28 or CD40-CD154 costimulatory pathways by using adenovirus vector coding CTLA4Ig (AdCTLA4Ig) or CD40Ig (AdCD40Ig) genes induced donor-specific tolerance in rat liver transplantation. In this study, we asked whether these gene-therapy-based costimulation blockade would induce tolerance in cardiac transplantation. METHODS: Heterotopic heart transplantation was performed in a full major histocompatibility complex (MHC) barrier combination of ACI (RT1avl) to Lewis (LEW, RT1l) rats. Vector (1 x 10(9) plaque forming unit [PFU]), AdLacZ, AdCTLA4Ig, or AdCD40Ig, was administered intravenously to recipient animals immediately after grafting, and graft survival, serum CTLA4Ig/CD40Ig levels, and graft histology were assessed. Tolerance was determined by secondary skin-graft challenging. RESULTS: Allografts of both untreated and AdLacZ controls were promptly rejected within 7 days, whereas a single treatment with AdCTLA4Ig or AdCD40Ig significantly prolonged median graft survival to 55.5 and 28.5 days, respectively. In contrast, the combined AdCTLA4Ig and AdCD40Ig gene therapy maintained high CTLA4Ig and CD40Ig levels through the posttransplant period and allowed long-term cardiac allograft survival for more than 270 days. However, both donor and third-party skin grafts were rejected in the animals who harbored cardiac grafts over 150 days. Also, typical features of chronic rejection were evident in the long-term surviving grafts. CONCLUSION: Simultaneous blockade of CD28 and CD154 pathways by AdCTLA4Ig plus AdCD40Ig induces a strong immunosuppression that allows long-term acceptance of full MHC mismatched cardiac graft in rats. This strategy, however, was not enough to induce tolerance to skin grafts and to avoid chronic rejection, as shown in the liver-transplantation model. PMID- 14557760 TI - Is selenium deficiency an important risk factor for chronic graft nephropathy? A pilot study. AB - BACKGROUND: Lipid peroxidation by free radicals is a key step in the development of atherosclerosis. Chronic graft nephropathy (CGN) is a common cause of allograft failure and shares many histologic features with atherosclerosis. Although hyperlipidemia is a common finding in renal transplant recipients, not all patients develop CGN. We hypothesized that the degree of damage sustained is related to recipient antioxidant status and that only those who are antioxidant deficient succumb to free radical attack and develop CGN. We aimed to determine the antioxidant profiles of patients with biopsy-proven CGN and to compare their profiles to transplant patients with good renal function. METHODS: Plasma selenium and vitamin A and E concentrations were measured in 10 patients with CGN and 10 contemporaneous, sex-matched patients with normal renal graft function, who received the same immunosuppressive therapy. RESULTS: Patients with CGN had significantly lower plasma selenium concentrations compared with those with normal renal allograft function (P<0.05). There were no significant differences in plasma vitamin A or E concentrations between the two groups. There was no difference in the prevalence of any of the immunologic or nonimmunologic risk factors: human leukocyte antigen mismatches, panel-reactive antibody status, number of rejection episodes, cold ischemic time, hyperlipidemia, hypertension, diabetes, and cytomegalovirus infection between the two groups. CONCLUSIONS: Patients with CGN have evidence of selenium deficiency, suggesting that impaired antioxidant status may contribute to the development of CGN. PMID- 14557761 TI - Increased expression of transforming growth factor-beta and eosinophil infiltration is associated with the development of transplant arteriosclerosis in long-term surviving cardiac allografts. AB - BACKGROUND: Transplant arteriosclerosis is a major limiting factor for long-term function of allografts in clinical transplantation. This study investigated the impact of three different protocols capable of inducing long-term allograft survival on the development of transplant arteriosclerosis and immune response in cardiac allografts. METHODS: CBA.Ca (H2k) recipients of fully allogeneic C57/BL10 (H2b) heart grafts received a short-term course of anti-CD154 antibody or were pretreated with anti-CD4 antibody in combination with donor alloantigen in the form of CBK (H2k+Kb) bone marrow or C57BL/10 donor-specific transfusion (DST). Grafts were analyzed on day 40 or 100 after transplantation for transplant arteriosclerosis and expression of interferon-gamma, interleukin (IL)-2, IL-4, IL 10, IL-12p40, inducible nitric oxide synthase, and transforming growth factor (TGF)-beta1 mRNA. Serum was analyzed for the presence of alloantibodies. RESULTS: Intimal proliferation was 62%+/-11% on day 40 in the anti-CD154 group, progressed from 31%+/-10% on day 40 to 68%+/-8% on day 100 in the CBK-bone marrow group, but remained stable at 39%+/-4% in the DST group. Increased transplant arteriosclerosis on day 100 was associated with high intragraft TGF-beta1 mRNA production and eosinophil infiltration, but not alloantibody production. Progressing transplant arteriosclerosis was associated with increased IL-4 expression. CONCLUSION: Treatment protocols for the induction of long-term allograft survival can differ substantially in the extent and kinetics of transplant arteriosclerosis. IL-4 and TGF-beta1 may be two potential therapeutic targets to attenuate the development of transplant arteriosclerosis in the long term. PMID- 14557762 TI - Porcine endothelium activated by anti-alpha-GAL antibody binding mediates increased human neutrophil adhesion under flow. AB - BACKGROUND: Neutrophils participate in acute vascular rejection (AVR) of organ xenografts. Induced antibodies (Abs), including anti-Galalpha1,3Gal (alpha-Gal) Abs, have been suggested to cause AVR. We investigated the adhesion of naive human neutrophils to porcine aortic endothelial cells (PAECs) stimulated with anti-alpha-Gal Abs under conditions of flow. In addition, the ability of human neutrophils to adhere to human and porcine endothelium under static and flow conditions was evaluated. METHODS AND RESULTS: In a flow-adhesion assay, a significant increase in adhesion of human neutrophils to PAECs, but not to human aortic endothelial cells (HAECs), was detected 6 hours after anti-alpha-Gal Ab binding. After Ab stimulation, PAECs expressed CD62E and increased levels of CD106, indicating an activated endothelial cell (EC) phenotype. In a migration assay, supernatants from Ab-stimulated PAECs induced migration of human neutrophils, which was partially blocked by anti-porcine (p) interleukin (IL)-8 Abs and an antagonist to platelet-activating factor (PAF). In static and flow adhesion assays, no difference in adhesion of human neutrophils to unstimulated or tumor necrosis factor (TNF)-alpha-stimulated HAECs and PAECs could be detected. CONCLUSIONS: Our data suggest that anti-alpha-Gal Abs play an important role in the initiation of AVR by mediating adhesion and recruitment of neutrophils within an organ xenograft. In contrast with previous investigations, our data argues against a differential recognition of PAECs and HAECs by human neutrophils. Thus, to prevent AVR and accomplish long-term xenograft survival, it will be important to remove anti-alpha-Gal Abs before and after pig-to-human transplantation. PMID- 14557763 TI - Five-year results of kidney transplantation under tacrolimus-based regimes: the persisting significance of vascular rejection. AB - BACKGROUND: Acute rejection has been the major risk factor for medium-term kidney graft loss because of chronic allograft nephropathy. We investigated whether the use of improved immunosuppression has altered the relationship between acute and chronic rejection by analyzing data from 245 renal transplant patients receiving Tacrolimus-based immunosuppression. RESULTS: Five-year graft survival (censored for death with functioning graft) was 88.8% with no significant difference between living and cadaveric kidney transplants. The only significant predictor of medium-term graft loss was acute vascular rejection. CONCLUSION: Under Tacrolimus-based immunosuppression, the occurrence of acute interstitial rejection, even when occurring late, repeatedly, or with failure of graft function to return to baseline, was not associated with chronic allograft nephropathy or medium-term graft loss. Vascular rejection remains the major immunological obstacle to long-term transplant success. Five-year overall survival rates with a functioning graft of 80% with 90% graft survival censored for death with function seem to be realistic and achievable goals. PMID- 14557765 TI - Trichilemmal carcinoma: a case report of a rare skin cancer occurring in a renal transplant patient. PMID- 14557764 TI - Intracytoplasmic tagging of cells with ferumoxides and transfection agent for cellular magnetic resonance imaging after cell transplantation: methods and techniques. AB - BACKGROUND: Superparamagnetic iron oxides (SPIO) are being used to label cells for in vivo monitoring by magnetic resonance imaging (MRI). The purpose of this study is to present protocols using SPIO and a polycationic transfection agent for magnetic labeling of cells as a basis for cellular MRI. METHODS: Various concentrations of ferumoxides (FE)-poly-l-lysine (PLL) complexes were used to magnetically label cells. Iron incorporation into cells along with cell viability and short- and long-term toxicity were evaluated. RESULTS: Rapidly growing cell suspension and adherent cells were effectively labeled by means of endocytosis into endosomes at low concentrations of FE (25 microg/mL media) and PLL (0.75 microg/mL media). Hematopoietic stem cells and lymphocytes required higher concentrations of PLL (1.5 microg/mL) in serum-free media during initial FE-PLL complex formation before labeling the cells in culture. Total iron concentration in cells depended on the cell type, concentration of FE-PLL complexes in media, cellular density, and incubation time. Iron concentrations after overnight incubation with given FE at 25 microg/mL media resulted in, for example, T cells being labeled with 1 to 3 pg/cell of intracytoplasmic endosomal iron and 15 to 20 pg/cell of intracytoplasmic iron in mesenchymal stem cells compared with 0.01 to 0.1 pg/cell for unlabeled cells. Protocols developed for this study demonstrated no adverse effect on the cell viability, functional capacity, or toxicity. CONCLUSION: This technique can be used to label cells for in vivo MRI tracking of stem cells and lymphocytes. FE at a concentration of 25 to 50 microg/mL with a ratio of SPIO to PLL of 1:0.03 to 1:0.06 would be sufficient to label cells for cellular MRI. PMID- 14557766 TI - Diffuse C4d deposition and morphology of acute humoral rejection in a stable renal allograft. PMID- 14557767 TI - Insulin independence after conversion to tacrolimus and sirolimus-based immunosuppression in islet-kidney recipients. PMID- 14557768 TI - Enlargement of the cadaveric-liver donor pool using in-situ split-liver transplantation despite complex hepatic arterial anatomy. PMID- 14557769 TI - Disseminated Penicillium marneffei infection in a renal-transplant recipient successfully treated with liposomal amphotericin B. PMID- 14557770 TI - In memoriam: Rupert E. Billingham (1921-2002). PMID- 14557773 TI - The magic's in the moment. AB - Learn to look at your own management highlights for inspiration. PMID- 14557772 TI - Beyond the charge nurse. PMID- 14557774 TI - Prepare for periodic performance review. AB - Evaluate your facility's compliance with Accreditation Participation Requirements. PMID- 14557775 TI - Today's challenges shape tomorrow's technology, Part 1. AB - ring's viability and progress, leaders must use the challenges facing the profession and the health care industry as a blueprint for a future technological vision. PMID- 14557776 TI - Standing up to the scrutiny of medical malpractice. AB - Correct charting, updated policies and procedures, and firm staff understanding are critical components of avoiding medical malpractice. PMID- 14557777 TI - Create accountable, balanced work environments. PMID- 14557778 TI - Innovative leadership through emotional intelligence. AB - Explore the relatively new concepts of emotional labor and emotional intelligence, and review leadership strategies that draw from these elements. PMID- 14557779 TI - When push comes to shove: managers as mediators. AB - Nurse executives build healthy environments by encouraging managers to embrace their role as mediators. PMID- 14557780 TI - Mediation, not litigation. PMID- 14557783 TI - What's behind RN turnover? AB - Examine the results of a qualitative study designed to enhance your understanding of RNs' perceptions of the factors prompting them to leave employment in acute care settings. PMID- 14557784 TI - Uncovering pressure ulcer incidence. AB - Researchers compare state-specific pressure ulcer incidence, staff mix, and nursing care quality outcomes data. PMID- 14557785 TI - Leading your leader. AB - Learn how one facility developed nurse managers' leadership skills through a questionnaire process and training opportunities. PMID- 14557786 TI - The Magnetic pull. AB - Learn how leading hospitals attained Magnet status from the American Nurses Credentialing Center. PMID- 14557787 TI - Where do nurse educators fit? PMID- 14557788 TI - Consider case management. AB - Case management serves as an alternative career for advanced practice nurses. PMID- 14557789 TI - Controlling food intake and energy balance: which macronutrient should we select? PMID- 14557790 TI - Neural systems controlling food intake and energy balance in the modern world. AB - PURPOSE OF REVIEW: Obesity continues to increase around the globe, and creates a major health problem because of its comorbidities such as diabetes and cardiovascular disease. Considering that the major cause of most human obesities is the modern lifestyle in a rapidly changing environment, this suggests that the battle is between brain areas controlling internal metabolic homeostasis and those dealing with cognitive and emotional processing of external information. Thus, this review is intended to bridge this apparent neural dichotomy and highlight possible ways of interactions between these homeostatic and nonhomeostatic systems. RECENT FINDINGS: Rapid progress has been made in identifying the molecular mechanisms underlying the neural circuits regulating food intake and energy balance. Specific populations of peptidergic neurons in the medial hypothalamus can be considered metabolic integrators sensing both short and long-term availability of fuels. These cell groups in turn connect with various other brain regions to orchestrate adaptive responses through changes in food intake, as well as endocrine and autonomic responses. Alternatively, the impact of the environmental changes on ingestive behavior have been documented, and many of the underlying cortico-limbic pathways are starting to be identified. It has been proposed that a switch from instinctual control of energy balance to a more cognitive control will be necessary to stop the obesity epidemic. SUMMARY: The realization that there is nothing wrong with the homeostatic regulatory system in common obesity suggests that a systematic investigation of pathways that link the 'cognitive' with the 'metabolic' brain might be a more promising area of research. Such an investigation would reveal new molecular and cellular mechanisms providing new drug targets, leading to behavioral therapies that are directed more towards the causes of common obesity. PMID- 14557791 TI - Peripheral signals in the control of satiety and hunger. AB - PURPOSE OF REVIEW: Food intake is critical for survival and is a complex behavior with multiple levels of control. Short-term, meal-related signals arise from many sources including the gastrointestinal tract, the environment, and higher centers in the brain. As described in this review, inputs from the gastrointestinal tract can exert potent effects on meal initiation, meal termination, and meal frequency. The complex array of signals generated from the gastrointestinal system and from adipose tissue, which participate in the regulation of food intake, and specifically how these signals relate to satiety and hunger, is the focus of this review. RECENT FINDINGS: Literature on the role of the well-studied gastrointestinal peptide, cholecystokinin, in satiety, in addition to its interaction with long-term adiposity signals in mediating food intake will be reviewed. In addition, literature on the gastrointestinal hormones glucagon-like peptide 1, apolipoprotein A-IV and peptide YY, and how they may act to regulate satiety, is described. Finally, the newly discovered hormone, ghrelin, and how it relates to meal initiation and hunger is discussed. SUMMARY: A better understanding of these systems and how they relate to body adiposity will prove to have important clinical applications. The available data suggest that interventions directed at multiple targets in the energy homeostasis system may be necessary to achieve and maintain weight loss. PMID- 14557792 TI - Fats and food intake. AB - PURPOSE OF REVIEW: Given the global rates of obesity and the potential link to dietary fat intake, understanding the role of fat in the regulation of food intake is critical. Some short-term, laboratory-based studies demonstrate poor compensation for manipulation of fat content, leading to passive overconsumption, while others demonstrate compensation to levels similar to other macronutrients. The observation of compensation in the short term does not concur with long-term rates of obesity increase. This review discusses factors that may explain at a physiological level these discrepancies, in particular fat structure, dietary adaptation, and palatability. RECENT FINDINGS: Medium-chain triglycerides have been demonstrated to be more satiating and promote weight loss. Recent data suggest different gastrointestinal transduction mechanisms elicit vagal afferent firing for fatty acids of different chain length. Dietary adaptation to fat can influence the sensitivity of the feedback response, which appears to be nutrient specific and relate to gastric emptying rates and hormonal feedback. Fat content has been found to influence palatability of foods. Recently it has been demonstrated that increasing palatability can partially override the satiating effects of covertly manipulated macronutrient preloads. Recent data suggest that hormonal influences may also affect the palatability response. SUMMARY: It is becoming increasingly clear that although energy density of diets is a major factor determining intake, macronutrient structure, subject, dietary and taste differences can all play an important modulatory influence on the final response on food intake. Further understanding of these factors and interactions may provide strategies to help aid weight regulation. PMID- 14557793 TI - The significance of protein in food intake and body weight regulation. AB - PURPOSE OF REVIEW: To highlight the underexposed but important role of protein in food intake and body weight regulation. RECENT FINDINGS: Protein plays a key role in food intake regulation through satiety related to diet-induced thermogenesis. Protein also plays a key role in body weight regulation through its effect on thermogenesis and body composition. A high percentage of energy from dietary protein limits body weight (re)gain through its satiety and energy inefficiency related to the change in body composition. SUMMARY: Protein is more satiating than carbohydrate and fat in the short term, over 24 h and in the long term. Thermogenesis plays a role in this satiety effect, but the role of satiety hormones still needs to be elucidated. On the short-term 'fast' proteins are more satiating than 'slow' proteins, and animal protein induces a higher thermogenesis than vegetable protein. In the longer term the higher postabsorptive satiety and thermogenesis are sustained irrespective of the protein source. High-protein diets affect body weight loss positively only under ad-libitum energy intake conditions, implying also a decreased energy intake. Body composition and metabolic profile are improved. Additional protein consumption results in a significantly lower body weight regain after weight loss, due to body composition, satiety, thermogenesis, and energy inefficiency, while the metabolic profile improves. Implications from these findings are: for practice, recommendations for increasing the percentage of energy from protein while reducing energy intake; for clinical research, assessment of the paradox of increasing the percentage energy from a highly satiating macronutrient; of the potential roles of protein in a negative and positive energy balance; assessment of possibilities of replacing dietary protein by effective amino acids or peptides that may show a similar impact on body weight regulation. PMID- 14557794 TI - Alcohol and food intake. AB - PURPOSE OF REVIEW: Alcohol is commonly consumed around mealtimes, and both the immediate pharmacological actions of ethanol and the energy generated by metabolism of alcohol have the potential to modify food intake. Effects of moderate alcohol consumption on food intake in humans will be reviewed, and potential mechanisms considered. RECENT FINDINGS: Unlike other macronutrients, there is minimal evidence for any reduction in food intake to compensate for the potential energy in alcohol. In contrast, moderate alcohol consumption prior to a test meal leads to a short-term increase in food intake. This stimulatory effect of alcohol is not apparent beyond acute administration, but the inability to reduce voluntary energy intake in response to energy from alcohol metabolism is evident over extended periods. Alcohol suppresses fatty acid oxidation, increases short-term thermogenesis and stimulates a number of neurochemical and peripheral systems implicated in appetite control, including inhibitory effects on leptin, glucagon-like peptide-1, and serotonin, and enhancement of gamma-aminobutyric acid, endogenous opioids and neuropeptide Y. All of these effects could lead to overeating, and mechanisms underlying appetite stimulation through alcohol require further substantiation. SUMMARY: Alcohol is a complex component of the diet, and appears to have multiple effects on appetite. Failure to reduce food intake in response to energy from alcohol makes moderate alcohol consumption a risk factor for obesity. Further integration of evidence from nutrition and neuroscience will be crucial to our understanding of effects of alcohol on appetite. PMID- 14557795 TI - The question of micronutrient supplements. PMID- 14557796 TI - Antimutagenic activity of tea: role of polyphenols. AB - PURPOSE OF REVIEW: Tea is considered to be one of the most promising dietary chemopreventive agents and, consequently, it is being studied extensively worldwide. Despite the fact that tea has proved very efficient in affording protection against chemical-induced cancer in animal models of the disease, epidemiological studies do not always support the laboratory findings, so that the value of tea as a human anticarcinogen may be considered as 'not proven'. A major mechanism of the anticarcinogenic activity of tea in animals is impairment of the interaction of carcinogens with DNA leading to mutations. The antimutagenic activity of tea as well as the underlying mechanisms will be reviewed, and the role of polyphenols, the postulated bioactive components, and caffeine will be critically evaluated. RECENT FINDINGS: In rats, exposure to tea modulated the disposition of heterocyclic amines, a major group of food-borne carcinogens, stimulating the pathways that lead to deactivation, and this is concordant with the established ability of tea to modulate the carcinogen metabolizing enzyme systems. These observations provide a rational mechanism for the anticarcinogenic activity of tea in animals. SUMMARY: The beneficial activities of tea have always been attributed to the polyphenols, as these are present in tea at substantial concentrations and are endowed with antioxidant activity. It is becoming increasingly evident, however, that the bioavailability of these compounds is poor as a result of limited absorption and presystemic metabolism by mammalian and microbial enzymes. We propose that the biological activity of tea may be mediated by caffeine and microbial metabolites of polyphenols. PMID- 14557797 TI - Tomato products and lycopene supplements: mandatory components in nutritional treatment of cancer patients? AB - PURPOSE OF REVIEW: This review presents the latest experimental and clinical research focussing on the relationship between the intake of tomato products and lycopene supplementation and carcinogenesis, with the aim of drawing conclusions for concepts of clinical nutritional support. RECENT FINDINGS: Apart from the preventative role of tomato products/lycopene intake there is evidence that oral supplementation of these compounds in cancer patients may also improve the biomarkers of carcinogenesis and reduce tumour growth. New experimental studies in animal models provide insights concerning the potential mechanism(s). SUMMARY: Although the first clinical trials are promising, it is too early to make final recommendations for nutritional therapy in cancer patients. Whether the bioactive compound in tomatoes is lycopene or whether other substrates contribute to the beneficial physiological effects is still unclear. PMID- 14557798 TI - Nutraceuticals--a pharmaceutical viewpoint: part II. AB - PURPOSE OF REVIEW: To review pharmaceutical and pharmacological issues relating to the benefits and risks associated with the use of naturally sourced nutraceuticals when administered singly or in combinations. RECENT FINDINGS: The application of vegetable extracts or dietary supplementation with selenium or antioxidant vitamins results in positive benefits on immunity and other phenomena associated with chronic diseases, ageing and cancer. However, there appear to be no cardiovascular benefits from vitamin mixtures, which may in fact cause harm. Therefore, although recent publications have increased our understanding of the metabolic actions of nutraceuticals, learning to use them to the best advantage is going to require products with uniform and consistent quality. Unfortunately, a single purified substance will not always have the same antioxidant activity, nor provide the same clinical benefits as nutraceutical mixtures and combinations occurring in natural extracts. In order to perform first-class clinical studies to determine safety and efficacy, the stability, compatibility and other pharmaceutical variables inherent in many of these combination products will have to be better controlled. SUMMARY: Nutraceuticals have potent biological actions. Their use is increasing dramatically, and there is growing evidence of clinical benefits. No medicinal products are completely safe so their risks need to be characterized and controlled. Imposing pharmaceutical levels of control and regulation would increase costs and reduce patient access to new products, but the evidence is compelling that closer monitoring of raw materials, processing and formulation will be required to maximize the benefits and minimize the risks. PMID- 14557799 TI - Using live, attenuated influenza vaccine for prevention and control of influenza: supplemental recommendations of the Advisory Committee on Immunization Practices (ACIP). AB - This report summarizes recommendations by the Advisory Committee on Immunization Practices (ACIP) for using intranasally administered, trivalent, cold-adapted, live, attenuated influenza vaccine (LAIV), which was approved for use in the United States on June 17, 2003 (FluMist trade mark, produced by MedImmune, Inc., Gaithersburg, Maryland). LAIV is currently approved for use among healthy persons (i.e., those not at high risk for complications from influenza infection) aged 5 49 years. This report includes information regarding 1) vaccine composition and mechanisms of action; 2) comparison between LAIV and trivalent inactivated influenza vaccine; 3) effectiveness and safety of LAIV; 4) transmission and stability of LAIV viruses; 5) recommendations and contraindications for using LAIV; and 6) dosage and administration of LAIV. This report supplements the 2003 ACIP recommendations regarding prevention and control of influenza (CDC. Prevention and Control of Influenza: Recommendations of the Advisory Committee on Immunization Practices [ACIP]. MMWR 2003;52[No. RR-8]:1-36.) PMID- 14557801 TI - Genomic instability and bystander effects: a historical perspective. AB - Data have been emerging over the past two decades concerning two phenomena in which important biological effects of ionizing radiation arise in cells that in themselves receive no radiation exposure. In the first, radiation-induced genomic instability, biological effects occur in the progeny of the irradiated cell after many generations of cell division. In the second, radiation-induced bystander effects, they arise in cells that receive no radiation exposure as a consequence of damage signals transmitted from neighboring irradiated cells; transmission may be mediated either by direct intercellular communication through gap junctions, or by factors released into the surrounding medium. In both phenomena, the biological effects appear to be associated with an upregulation of oxidative metabolism. The present paper is designed to review the historical background leading to our current knowledge of these two phenomena, and to indicate some future directions for research that will allow us to assess better their importance in the health effects of exposure to ionizing radiation. PMID- 14557802 TI - Radiation-induced DNA damage and delayed induced genomic instability. AB - Ionizing radiation induces genomic instability, which is transmitted over many generations after irradiation through the progeny of surviving cells. Induced genomic instability is manifested as the expression of the following delayed effects: delayed reproductive death or lethal mutation, chromosomal instability, and mutagenesis. Since induced genomic instability accumulates gene mutations (actually genomic instability is the process whereby gene mutation increases subtle difference) and gross chromosomal rearrangements, it has been thought to play a role in radiation-induced carcinogenesis. Radiation-induced genomic instability exerts its effects for prolonged periods of time, suggesting the presence of a mechanism by which the initial DNA damage in the surviving cells is memorized. Recent studies have shown that such memory transmission causes delayed DNA breakage, which in turn plays a role in the induction of delayed phenotypes. Although radiation-induced genomic instability has been studied for years, many questions remain to be answered. This review summarizes the current data on radiation-induced genomic instability. In particular, the mechanism(s) involved in the initiation and perpetuation of radiation-induced genomic instability, and a role of delayed activation of p53 protein are discussed. PMID- 14557803 TI - Role of genetic background in induced instability. AB - Genomic instability is effectively induced by ionizing radiation. Recently, evidence has accumulated supporting a relationship between genetic background and the radiation-induced genomic instability phenotype. This is possibly due to alterations in proteins responsible for maintenance of genomic integrity or altered oxidative metabolism. Studies in human cell lines, human primary cells, and mouse models have been performed predominantly using high linear energy transfer (LET) radiation, or high doses of low LET radiation. The interplay between genetics, radiation response, and genomic instability has not been fully determined at low doses of low LET radiation. However, recent studies using low doses of low LET radiation suggest that the relationship between genetic background and radiation-induced genomic instability may be more complicated than these same relationships at high LET or high doses of low LET radiation. The complexity of this relationship at low doses of low LET radiation suggests that more of the population may be at risk than previously recognized and may have implications for radiation risk assessment. PMID- 14557804 TI - Mouse models for induced genetic instability at endogenous loci. AB - Exposure to environmental factors and genetic predisposition of an individual may lead individually or in combination to various genetic diseases including cancer. These diseases may be a consequence of genetic instability resulting in large scale genomic rearrangements, such as DNA deletions, duplications, and translocations. This review focuses on mouse assays detecting genetic instability at endogenous loci. The frequency of DNA deletions by homologous recombination at the pink-eyed unstable (p(un)) locus is elevated in mice with mutations in ATM, Trp53, Gadd45, and WRN genes and after exposure to carcinogens. Other quantitative in vivo assays detecting loss of heterozygosity events, such as the mammalian spot assay, Dlb-1 mouse and Aprt mouse assays, are also reviewed. These in vivo test systems may predict hazardous effects of an environmental agent and/or genetic predisposition to cancer. PMID- 14557805 TI - Clinical implications of radiation-induced genomic instability. AB - Radiation-induced genomic instability encompasses a range of measurable end points such as chromosome destabilization, sister chromatid exchanges, gene mutation and amplification, late cell death and aneuploidy, all of which may be causative factors in the development of clinical disease, including carcinoma. Clinical implications of genomic instability can be broadly grouped into two main areas: as a marker for increased cancer risk/early detection, and as a consequence of radiation therapy (IR) that may be causative of, or a strong marker for, the induction of a therapy-induced second malignancy. Research in human populations has been limited, but broadly encompasses three populations: those exposed to alpha-particle irradiation, those with a cancer diagnosis who were examined for lymphocyte sensitivity to IR as a biomarker for risk of cancer induction, and those who successfully completed radiation therapy for an index cancer and who were examined for the induction of a second malignancy. This review examines each of those populations in turn and offers some potential future research directions to better elucidate the role of radiation-induced genomic instability in clinical disease. PMID- 14557806 TI - Second cancers after radiotherapy: any evidence for radiation-induced genomic instability? AB - Do second primary cancers in humans arise from radiation-induced somatic genomic instability after radiotherapy for the first malignancy? The amount of truly pertinent human information on this issue is sparse, leading to the conclusion that we cannot confirm or refute that instability induction by radiation is involved. However, the in vitro findings of radiation-induced genomic instability through bystander effects or increased mutation rates in cell progeny of apparently normal but irradiated cells are provocative and their transferability to human in vivo biology deserves further investigation. We describe possible animal and human studies to stimulate ideas, but the collaborative commitment of multiple large institutions to tumor tissue procurement and retrieval will be essential. In addition, detecting the temporal progression of genomic instability and identifying the salient genetic events as being radiation-induced will be pivotal. Execution of some of the studies suggested is not possible now, but applying next-generation methods could bring the concepts to fruition. As nearly one in 10 cancer diagnoses are second (or higher) malignancies, it is important to understand the contribution of radiotherapy to second cancer induction and pursue well-coordinated efforts to determine the role of induced genomic instability. PMID- 14557807 TI - Radiation-induced bystander effects, carcinogenesis and models. AB - Implications for carcinogenesis of radiation-induced bystander effects are both mechanistic and practical. They include induction of second cancers, perturbations to tissue social control and induction of genomic instability and delayed or immediate mutations in areas not receiving a direct deposition of energy. Bystander effects have consequences for DNA damage-mutation-cancer initiation paradigms of radiation carcinogenesis that provide the mechanistic justification for low-dose risk estimates. If carcinogenesis does not result from directly induced DNA mutations, then the carcinogenic initiation process may not simply relate to radiation dose. Modification of the preclonal state through genetic and epigenetic mechanisms may occur. To deal with the complexity of these interactions, a 'chaotic' or 'bifurcation' model invoking autopoietic theory is proposed that could accommodate both beneficial (hormetic) and harmful effects of radiation at comparable doses. Carcinogenesis may then be thought of as the result of a disturbance of the genetic/epigenetic balance occurring within the organ. Ultimate clonality may reflect domination due to selection processes rather than the initiating damage. PMID- 14557808 TI - Genomic instability and bystander effects induced by high-LET radiation. AB - An understanding of the radiobiological effects of high-linear energy transfer (LET) radiation is essential for radiation protection and human risk assessment. Ever since the discovery of X-rays was made by Rontgen more than a century ago, it has always been accepted that the deleterious effects of ionizing radiation, such as mutation and carcinogenesis, are due mainly to direct damage to DNA. With the availability of a precision single-particle microbeam, it is possible to demonstrate, unequivocally, the presence of a bystander effect with many biological end points. These studies provide clear evidence that irradiated cells can induce a bystander mutagenic response in neighboring cells not directly traversed by alpha-particles, and that cell-cell communication processes play a critical role in mediating the bystander phenomenon. Following exposure to high LET radiation, immortalized human bronchial (BEP2D) and breast (MCF-10F) cells have been shown to undergo malignant transformation through a series of successive steps, before becoming tumorigenic in nude mice. There is a progressive increase in genomic instability, determined either by gene amplification or allelic imbalance, with the highest incidence observed among established tumor cell lines, relative to transformed, nontumorigenic and control cell lines. PMID- 14557809 TI - Bystander responses induced by low LET radiation. AB - Radiation-induced bystander responses are observed when cells respond to their neighbours being irradiated. Considerable evidence is now available regarding the importance of these responses in cell and tissue models. Most studies have utilized two approaches where either a media-transferable factor has been assessed or cells have been exposed to low fluences of charged particles, where only a few percent are exposed. The development of microbeams has allowed nontargeted responses such as bystander effects to be more carefully analysed. As well as charged particle microbeams, X-ray microprobes have been developed, and several groups are also developing electron microbeams. Using the Gray Cancer Institute soft X-ray microprobe, it has been possible to follow the response of individual cells to targeted low doses of carbon-characteristic soft X-rays. Studies in human fibroblasts have shown evidence of a significant radiation quality-dependent bystander effect, measured as chromosomal damage in the form of micronuclei which is radiation quality dependent. Other studies show that even under conditions when only a single cell is targeted with soft X-rays, significant bystander-mediated cell killing is observed. The observation of bystander responses with low LET radiation suggests that these may be important in understanding radiation risk from background levels of radiation, where cells observe only single electron track traversals. Also, the indirect evidence for these responses in vivo indicates that they may have a role to play in current radiotherapy approaches and future novel strategies involving modulating nontargeted responses. PMID- 14557810 TI - Oxidative metabolism, gap junctions and the ionizing radiation-induced bystander effect. AB - Evidence accumulated over the past two decades has indicated that exposure of cell populations to ionizing radiation results in significant biological effects occurring in both the irradiated and nonirradiated cells in the population. This phenomenon, termed the 'bystander response', has been shown to occur both in vitro and in vivo and has been postulated to impact both the estimation of risks of exposure to low doses/low fluences of ionizing radiation and radiotherapy. Several mechanisms involving secreted soluble factors, oxidative metabolism and gap-junction intercellular communication have been proposed to regulate the radiation-induced bystander effect. Our current knowledge of the biochemical and molecular events involved in the latter two processes is reviewed in this article. PMID- 14557811 TI - Radiation-induced genomic instability and bystander effects: inter-related nontargeted effects of exposure to ionizing radiation. AB - The paradigm of genetic alterations being restricted to direct DNA damage after exposure to ionizing radiation has been challenged by observations in which cells that are not exposed to ionizing radiation exhibit responses typically associated with direct radiation exposure. These effects are demonstrated in cells that are the descendants of irradiated cells (radiation-induced genomic instability) or in cells that are in contact with irradiated cells or receive certain signals from irradiated cells (radiation-induced bystander effects). There is accumulating evidence that radiation-induced genomic instability may be a consequence of, and in some cell systems may also produce, bystander interactions involving intercellular signalling, production of cytokines and free-radical generation. These processes are also features of inflammatory responses that are known to have the potential for both bystander-mediated and persisting damage as well as for conferring a predisposition to malignancy. Thus, radiation-induced genomic instability and untargeted bystander effects may reflect inter-related aspects of inflammatory-type responses to radiation-induced stress and injury and contribute to the variety of pathological consequences of radiation exposures. PMID- 14557812 TI - Raft ceramide in molecular medicine. AB - Ceramide, generated by the action of acid sphingomyelinase (ASM), has emerged as a biochemical mediator of stimuli as diverse as ionizing radiation, chemotherapy, UVA light, heat, CD95, reperfusion injury, as well as infection with some pathogenic bacteria and viruses. ASM activity is also crucial for developmental programmed cell death of oocytes by apoptosis. Recently, we proposed a comprehensive model that might explain these diverse functions of ceramide: Upon contacting the relevant stimuli, ASM translocates into and generates ceramide within distinct plasma membrane sphingolipid-enriched microdomains termed rafts. Ceramide, which manifests a unique biophysical property, the capability to self associate through hydrogen bonding, provides the driving force that results in the coalescence of microscopic rafts into large-membrane macrodomains. These structures serve as platforms for protein concentration and oligomerization, transmitting signals across the plasma membrane. Preliminary data suggest that manipulation of ceramide metabolism and/or the function of ceramide-enriched membrane platforms may present novel therapeutic opportunities for the treatment of cancer, degenerative disorders, pathogenic infections or cardiovascular diseases. PMID- 14557813 TI - Induced genomic instability in irradiated germ cells and in the offspring; reconciling discrepancies among the human and animal studies. AB - Many studies confirmed that radiation induces genomic instability in whole-body systems. However, the results of the studies are not always consistent with each other. Attempts are made in the present review to resolve the discrepancies. Many of the studies in human and experimental animals utilize the length change mutation of minisatellite sequences as a marker of genomic instability. Minisatellite sequences frequently change their length, and the data obtained by conventional Southern blotting give rather qualitative information, which is sometimes difficult to scrutinize quantitatively. This is the problem inevitably associated with the study of minisatellite mutations and the source of some conflicts among studies in humans and mice. Radiation induction of genomic instability has also been assessed in whole-body experimental systems, using other markers such as the mouse pink-eyed unstable allele and the specific pigmentation loci of medaka fish (Oryzias latipes). Even though there are some contradictions, all these studies have demonstrated that genomic instability is induced in the germ cells of irradiated parents, especially of males, and in offspring born to them. Among these, transmission of genomic instability to the second generation of irradiated parents is limited to the mouse minisatellite system, and awaits further clarification in other experimental systems. PMID- 14557814 TI - Radiation-induced transgenerational instability. AB - To date, the analysis of mutation induction has provided an irrefutable evidence for an elevated germline mutation rate in the parents directly exposed to ionizing radiation and a number of chemical mutagens. However, the results of numerous publications suggest that radiation may also have an indirect effect on genome stability, which is transmitted through the germ line of irradiated parents to their offspring. This review describes the phenomenon of transgenerational instability and focuses on the data showing increased cancer incidence and elevated mutation rates in the germ line and somatic tissues of the offspring of irradiated parents. The possible mechanisms of transgenerational instability are also discussed. PMID- 14557815 TI - Is there a common mechanism underlying genomic instability, bystander effects and other nontargeted effects of exposure to ionizing radiation? AB - A number of nontargeted and delayed effects associated with radiation exposure have now been described. These include radiation-induced genomic instability, death-inducing and bystander effects, clastogenic factors and transgenerational effects. It is unlikely that these nontargeted effects are directly induced by cellular irradiation. Instead, it is proposed that some as yet to be identified secreted factor can be produced by irradiated cells that can stimulate effects in nonirradiated cells (death-inducing and bystander effects, clastogenic factors) and perpetuate genomic instability in the clonally expanded progeny of an irradiated cell. The proposed factor must be soluble and capable of being transported between cells by cell-to-cell gap junction communication channels. Furthermore, it must have the potential to stimulate cellular cytokines and/or reactive oxygen species. While it is difficult to imagine a role for such a secreted factor in contributing to transgenerational effects, the other nontargeted effects of radiation may all share a common mechanism. PMID- 14557816 TI - Dissecting HIV-1 through RNA interference. PMID- 14557817 TI - Cytostatic and apoptotic actions of TGF-beta in homeostasis and cancer. PMID- 14557818 TI - EB1 reveals mobile microtubule nucleation sites in Arabidopsis. AB - In plants, it is unclear how dispersed cortical microtubules are nucleated, polarized and organized in the absence of centrosomes. In Arabidopsis thaliana cells, expression of a fusion between the microtubule-end-binding protein AtEB1a and green fluorescent protein (GFP) results in labelling of spindle poles, where minus ends gather. During interphase, AtEB1a-GFP labels the microtubule plus end as a comet, but also marks the minus end as a site from which microtubules can grow and shrink. These minus-end nucleation sites are mobile, explaining how the cortical array can redistribute during the cell cycle and supporting the idea of a flexible centrosome in plants. PMID- 14557819 TI - Actin filament uncapping localizes to ruffling lamellae and rocketing vesicles. AB - Regulated actin filament assembly is critical for eukaryotic cell physiology. Actin filaments are polar structures, and those with free high affinity or barbed ends are crucial for actin dynamics and cell motility. Actin filament barbed-end capping proteins inhibit filament elongation after binding, and their regulated disassociation is proposed to provide a source of free filament ends to drive processes dependent on actin polymerization. To examine whether dissociation of actin filament capping proteins occurs with the correct spatio-temporal constraints to contribute to regulated actin assembly in live cells, I measured the dissociation of an actin capping protein, gelsolin, from actin in cells using a variation of fluorescence resonance energy transfer (FRET). Uncapping was found to occur in cells at sites of active actin assembly, including protruding lamellae and rocketing vesicles, with the correct spatio-temporal properties to provide sites of actin filament polymerization during protrusion. These observations are consistent with models where uncapping of existing filaments provides sites of actin filament elongation. PMID- 14557821 TI - The receptors and role of angiotensin II in knee joint blood flow regulation and role of nitric oxide in modulation of their function. AB - OBJECTIVES: Angiotensin-converting enzyme (ACE) upregulation in the stroma cells of arthritis rheumatoid joints may produce a higher tissue concentration of angiotensin II (angII), which is a vasoconstrictor and mitogen factor that causes local hypoxia and synovial proliferation. No study in the literature has examined the role of angII in joint blood flow (JBF) regulation and the potential effect of ACE inhibitors on JBF. METHODS: The study was performed on 20 Dutch white rabbits to examine the JBF response to angII, angII receptor subtypes, and the role of nitric oxide (NO) in angII effects in knee joint blood vessels. Drugs were administered locally through retrograde saphenous artery cannulation. Joint vascular resistance (JVR) was calculated by dividing the arterial blood pressure by the JBF. RESULTS: AngII increased JVR dose dependently. The angII type 1 (AT(1)) receptor antagonist losartan did not change the basal JVR but completely blocked the effect of angII on JVR. N(omega)-nitro-L-arginin methyl ester (L NAME) increased JVR by a mean (+/-SEM) of 25.8 +/- 8.7% (p < 0.05) but did not affect the joint vessel response to angII and losartan. CONCLUSIONS: AngII receptors are from the AT(1) subtype in normal joint blood vessels, but angII plays no significant role in JBF regulation. The basal release of NO plays a role in resting JBF regulation, but NO does not affect the AT(1) receptor-mediated vasoconstriction of joint blood vessels. PMID- 14557822 TI - Early hepatic microvascular injury in response to acetaminophen toxicity. AB - OBJECTIVE: The hepatic toxic response to acetaminophen (APAP) is characterized by centrilobular (CL) necrosis preceded by hepatic microvascular injury and congestion. The present study was conducted to examine changes in liver microcirculation after APAP dosing. METHODS: Male C57Bl/6 mice were treated with APAP (600 mg/kg body weight) by oral gavage. The livers of anesthetized mice were examined using established in vivo microscopic methods at 0, 0.5, 1, 2, 4, 6, 12 hours after APAP. RESULTS: The levels of hepatic transaminases (i.e., alanine aminotransferase [ALT] and aspartate transaminase) increased minimally for up to 2 hours. Thereafter, their levels were significantly and progressively increased. The numbers of swollen sinusoidal endothelial cells (SECs) in periportal regions were increased (3.5-fold) from 0.5 to 6 hours, and those in CL regions were increased (4.0-fold) at 0.5 and 1 hour. The intensity of in vivo staining for formaldehyde-treated serum albumin, which is a specific ligand for SECs, was reduced from 2 to 12 hours. Erythrocytes infiltrated into the space of Disse as early as 2 hours, and the area occupied by these cells was markedly increased at 6 hours. Sinusoidal perfusion was reduced from 1 through 12 hours, with a nadir (35% decrease) at 4 and 6 hours. Phagocytic Kupffer cell activity was significantly elevated from 0.5 through 12 hours. Although gadolinium chloride minimized the changes in sinusoidal blood flow and reduced ALT levels 6 hours after APAP, it failed to inhibit endothelial swelling, extravasation of erythrocytes, and CL parenchymal necrosis. CONCLUSIONS: These results confirm that APAP-induced SEC injury precedes hepatocellular injury, supporting the hypothesis that SECs are an early and direct target for APAP toxicity. These findings also suggest that reduced sinusoidal perfusion and increased Kupffer cell activity contribute to the development of APAP-induced liver injury. PMID- 14557823 TI - Hydrodynamic interactions between rolling leukocytes in vivo. AB - OBJECTIVE: The aim of this study was to characterize the hydrodynamic interactions between rolling and free-stream leukocytes in an in vivo model of selectin-mediated rolling, and to identify those physical mechanisms that influence the dynamics of transient adhesion with the walls of postcapillary venules. METHODS: Postcapillary venules of diameter 22-37 microm in the cheek pouch of anesthetized hamsters were visualized using intravital microscopy, with selectin-mediated rolling occurring in response to surgical preparation. RESULTS: Rolling velocity was found to be a strong function of the center-to-center separation with the nearest cell, and also was found to correlate strongly with the number of nearby cells. These effects are shown to be beyond that attributable to variations along the length of the vessel. Adherent leukocytes were observed to provide a nucleation site, precipitating further adhesion events of free-stream cells. CONCLUSIONS: The dynamics of the transient adhesion of leukocytes to the vessel wall in postcapillary venules is strongly dependent on the local concentration of adherent leukocytes, due to the complex hydrodynamics induced by their presence. The results are shown to agree well with theoretical considerations of the flow field induced by multiple nearby cells, suggesting a need for the future exploration of multicellular effects in the microcirculation. PMID- 14557824 TI - Role of the actin cytoskeleton in regulating endothelial permeability in venules. AB - OBJECTIVE: This study was performed to determine the effect of myosin light chain kinase (MLCK) inhibition on histamine- and thrombin-induced venular permeability in the rat mesentery, coincidental with actin cytoskeleton changes. METHODS: The mesenteric microvasculature of rats was perfused with a fluorescent tracer plus thrombin, histamine, or buffered saline, and the preparation was suffused with the MLCK inhibitor ML-7. The microvasculature then was stained for actin. RESULTS: The average (+/- SE) number of leaks per micrometer of venule length of the thrombin plus 5 microM ML-7 treatment (35.3 +/- 5.9 x 10(-4); n = 224) was significantly lower than that for the thrombin-only treatment (61.7 +/- 5.6 x 10( 4); n = 385; p < 0.001). The histamine preparations required higher concentrations of ML-7 to significantly reduce the number of leaks. A concentration of 100 microM reduced the average leak number from 20.8 +/- 3.9 x 10(-4) (n = 140) to 2.5 +/- 0.8 x 10(-4) (n = 383; p < 0.001), but 20 microM ML-7 had no effect. Although leaky areas of both the thrombin- and histamine-treated preparations showed disruptions of the peripheral actin rim coincident with fluorescein isothiocyanate-bovine serum albumin leaks, qualitative and quantitative differences were identified. CONCLUSIONS: The results suggest both similar and dissimilar mechanisms for thrombin and histamine regarding in situ endothelial gap formation. PMID- 14557825 TI - Hepatic microcirculatory dysfunction during cholestatic liver injury in rats. AB - OBJECTIVE: : The present study was conducted to elucidate the sequential alterations in the hepatic microvascular inflammatory response to extrahepatic biliary obstruction. METHODS: : The hepatic microvasculature in anesthetized Sprague-Dawley rats was studied by in vivo microscopy 3, 7, and 14 days after bile duct ligation (BDL) or sham operation. RESULTS: : The numbers of adhering leukocytes and swollen sinusoidal endothelial cells were significantly increased at 3, 7, and 14 days after BDL when compared with sham-operated controls. Concomitantly, the numbers of sinusoids containing blood flow were significantly and progressively decreased by up to 30%. The phagocytic activity of hepatic macrophages was significantly elevated during the development of biliary cholestasis. In particular, centrilobular phagocytosis at 14 days after BDL was significantly increased 1.4- to 2.0-fold when compared with that at 3 and 7 days after BDL. Electron microscopy also revealed evidence of activated Kupffer cells reflected by numerous filopodia and ruffles. CONCLUSIONS: : These results suggest that hepatic microcirculatory dysfunction subsequent to BDL contributes to cholestatic liver injury. Microcirculation (2003) 10, 421-432. doi:10.1038/sj.mn.7800208 PMID- 14557826 TI - The influence of probe fiber distance on laser Doppler perfusion monitoring measurements. AB - Laser Doppler perfusion monitoring (LDPM) is a noninvasive technique for monitoring skin microcirculation. The aim of this article was to investigate the influence of fiber separation on clinical LDPM measurements. A dual-channel LDPM system was used in combination with a probe that consists of two sets of detection fibers, at 0.2 and 1.0 mm from the illuminating fiber. Measurements were performed at the big toe of 8 healthy subjects and 11 subjects who had vascular disorders. In most cases, fluxes detected at both fiber distances showed very similar fluctuations. For each fiber separation, flux values of healthy subjects and patients were not significantly different. Furthermore, skin temperature (range: 22-34 degrees C) influenced the toe's pulp microcirculation markedly, increasing similarly at both probe separations, with a higher flux at a separation of 1.0 mm than at 0.2 mm. The flux ratio signal, obtained by dividing the flux at 0.2 mm by the flux at 1.0 mm, was significantly different between the two groups (p &< 0.05). In conclusion, the flux detected in vivo by means of LDPM, is influenced by the distance between the optical fibers. Use of the flux ratio with a multiseparation probe deserves attention as it is a possible marker for discriminating normal tissue perfusion from pathological skin tissue perfusion, independently from tissue temperature. PMID- 14557827 TI - Modulation of susceptibility to weight gain and insulin resistance in low birthweight rats by treatment of their mothers with leptin during pregnancy and lactation. AB - OBJECTIVES: To investigate whether administration of leptin to rats during pregnancy and lactation affects placental 11beta-hydroxysteroid dehydrogenase (11beta-HSD2) activity and the susceptibility of their offspring to weight gain and insulin resistance. DESIGN: Pregnant rats fed on a low-protein diet were administered leptin or saline by subcutaneous minipump from day 14 of gestation and throughout lactation. A further group was fed a normal diet and given saline. After weaning, the offspring of each group were fed on a normal diet until 6 weeks of age and then half of each group was transferred to a high-fat diet until 12 months of age. RESULTS: Plasma leptin levels were raised two-fold on days 16 18 of pregnancy in the leptin-treated dams, but, despite a constant rate of infusion, at parturition they dipped to control levels before rising again. The activity of placental 11beta-HSD2 was reduced by the low-protein diet; this reduction was prevented by treating the dams with leptin. The male offspring of the saline-treated dams gained more weight and had higher plasma leptin levels on the high fat than the chow diet, but the offspring of the leptin-treated dams did not. Fasting blood glucose and intraperitoneal glucose tolerance at 6 and 12 months of age was unaffected by the high-fat diet, but only the offspring of the leptin-treated dams achieved this control without raised insulin levels. CONCLUSIONS: The rate of leptin clearance appears to increase at parturition. The administration of leptin to rats during late pregnancy and lactation makes their male offspring less susceptible to high-fat-diet-induced weight gain and insulin resistance. PMID- 14557828 TI - Relationship between ambulation and obesity in older persons with and without low back pain. AB - CONTEXT: For obese older persons, ambulation is both functionally important and a means of weight control. The relationship between weight and ambulation is not known in this population. Also, the extent to which pain interferes with ambulation is not studied. OBJECTIVE: To examine the relationship between obesity and ambulation, and to determine the effect of pain and body mass index (BMI) on ambulation in older persons. DESIGN, SETTING, AND PARTICIPANTS: Retrospective study of 82 older persons, ages 55-79 y, some with no back pain recruited from the community, others with back pain or spinal stenosis recruited from a magnetic resonance imaging (MRI) scanner as part of a larger university study of spinal stenosis. OUTCOME MEASURES: Age, Visual Analog Scales for pain, BMI, patient diagnosis (no pain, mechanical back pain, and spinal stenosis), walking velocity and stride length on a 15-min laboratory ambulation test, and 1-week community ambulation measured with a pedometer (steps, distance, and energy expenditure). RESULTS: BMI had a significant inverse relationship with ambulatory measurements in terms of the distance walked, steps taken, and walking velocity. Pain severity and pain category also had a significant inverse relationship with these measures. A negative correlation was observed between pain and obesity, although the relationship was statistically nonsignificant. DISCUSSION: Obese older people walked less than the nonobese older people. Pain was associated with decreased ambulation. Clinicians who intend to encourage increased ambulation in older obese persons should consider possible barriers posed by musculoskeletal pain. PMID- 14557829 TI - Obesity is a chronic, relapsing neurochemical disease. AB - The debate about whether obesity should be called a disease continues. From a clinical perspective, it meets the criteria needed to call it a disease. It has an etiology--an imbalance between energy intake and expenditure. It has a pathogenesis in the feedback systems involving leptin, neurochemicals in the brain, and the neural and endocrine messages that respond to the intake of food. The pathology of obesity lies in its enlarged fat cells, and the pathophysiology lies in the changes in the secretion of products from these enlarged fat cells, including cytokines, procoagulants, inflammatory peptides, and angiotensinogen. These secretory products of fat cells and the increased mass of fat are responsible for the associated metabolic diseases, such as diabetes, hypertension, heart disease, sleep apnea, and some sorts of cancer. Treatments consist of techniques to alter the balance between energy intake and energy expenditure. This constellation of factors leads to the conclusion that obesity should be called a disease. PMID- 14557830 TI - Correlates of obesity in postmenopausal women with breast cancer: comparison of genetic, demographic, disease-related, life history and dietary factors. AB - BACKGROUND: Obesity in women has been associated with a variety of factors, including genetic predisposition, social class, early age at menarche, exercise, alcohol consumption and diet. Obesity is a risk factor for the occurrence and the recurrence of breast cancer in postmenopausal women, perhaps because of increased exposure to estrogen, insulin and insulin-like growth factors (IGFs). The progesterone receptor (PR) and the steroid hormone receptor coactivator pCIP/ACTR/AIB1/TRAM1/RAC3 (AIB1) are hypothesized to mediate signaling crosstalk between these hormonal pathways. Polymorphisms in both genes have been described and their association with breast cancer risk reported. If genetic factors contribute to obesity, and the PR and AIB1 genes influence estrogenic, insulin and IGF pathways, then genetic patterns resulting from PR and AIB1 polymorphisms may be associated with obesity in postmenopausal women. OBJECTIVE: We compared the PR and AIB1 genotypes of postmenopausal women with breast cancer with demographic, disease-related, reproductive, lifestyle and dietary variables in terms of the strength of their relationship with obesity (BMI> or =30 kg/m2). SUBJECTS: A total of 301 postmenopausal women previously diagnosed with Stage I, II or IIIA breast cancer, who are enrolled in the Women's Healthy Eating and Living (WHEL) study (age: 34.5-70.8 y, BMI: 17.8-54.6 kg/m2). MEASUREMENTS: The PR polymorphism PROGINS was identified by PCR. The length of the AIB1 polyglutamine repeat was determined by PCR and nondenaturing gel electrophoresis or DNA sequencing. BMI was obtained at the baseline clinic visit upon entry into the WHEL study. Information about date of diagnosis, stage of disease, tumor hormone receptor status and adjuvant treatment received were obtained from medical records. Reproductive, menstrual history, demographic, family history of cancer, smoking history and exercise frequency and intensity information were obtained from questionnaires. Dietary and alcohol intake data came from four 24-h telephone recalls of food intake obtained at the study entry. RESULTS: The combined inheritance of PROGINS A1/A1 and AIB1 28/29, 28/30, 28/31, 29/29 or 29/30 (AIB1 LG) genotypes (adjusted odds ratio (OR)=2.22 (95% confidence interval 1.25-3.93)) and early age at menarche (<12 y) (adjusted OR=2.34 (1.12-4.86)) were each associated with the risk for obesity. Current use of tamoxifen (adjusted OR=0.49 (0.28-0.87)) and an alcohol intake > or =10 g/day (adjusted OR=0.28 (0.11 0.77)) were inversely associated with BMI > or =30 kg/m2. CONCLUSION: Early age at menarche and a PROGINS A1/A1+AIB1 LG genetic pattern had comparable levels of association with obesity in this cross-sectional sample of postmenopausal women with breast cancer. Since this was a cross-sectional rather than a case-control design, the association between PROGINS and AIB1 genotype and obesity found in this sample should be considered preliminary, and must be re-evaluated with a new and larger sample. PMID- 14557831 TI - Expression of the thermogenic nuclear hormone receptor coactivator PGC-1alpha is reduced in the adipose tissue of morbidly obese subjects. AB - Peroxisome proliferator-activated receptor gamma coactivator 1alpha (PGC1alpha) is an accessory protein which can potentiate the transcriptional activation function of many nuclear hormone receptors. Its tissue distribution and physiological studies suggest that its principal in vivo roles are to promote cold-induced thermogenesis, mitochondrial biogenesis, hepatic gluconeogenesis, and fatty acid beta-oxidation. It is expressed in the white adipose tissue of both humans and rodents, and in rodents it has been suggested to mediate in part the leptin-induced conversion of white adipocytes from fat storing to fat oxidising cells. In this study, quantitative real-time PCR has been used in human tissue to demonstrate that (1) PGC1alpha mRNA levels in subcutaneous fat are three-fold lower in morbidly obese than in slim subjects; (2) there are no differences in PGC1alpha mRNA between omental and subcutaneous mature adipocytes; (3) there is a robust induction of PGC1alpha expression during subcutaneous human preadipocyte differentiation ex vivo. Whether low PGC1alpha expression is a prelude to the development of obesity, or a consequence of that obesity, attempts to upregulate endogenous white adipose tissue expression may prove a valuable new avenue to explore in obesity therapy. PMID- 14557832 TI - Overweight Japanese with body mass indexes of 23.0-24.9 have higher risks for obesity-associated disorders: a comparison of Japanese and Mongolians. AB - OBJECTIVE: The degree of obesity of Asians is less than that of Caucasians. It has been suggested that Japanese, categorized as having normal weight (BMI<25.0), as defined by WHO (2000), have a tendency toward increased incidences of dyslipidemia and diabetes. Our objective was to analyze parameters constituting obesity-associated disorders in overweight Japanese and Mongolians with a body mass index (BMI) of 23.0-24.9, and to assess the suitability for Asians of the Regional Office for Western Pacific Region of WHO criteria pertaining to obesity (WPRO criteria, 2000). DESIGN: Cross-sectional study in a workplace setting. SUBJECTS: A total of 386 Japanese men and 363 Japanese women, and 102 Mongolian men and 155 Mongolian women. MEASUREMENTS: Anthropometric measurements (weight, height, waist circumference, hip circumference and blood pressure) and metabolic measurements (plasma levels of total cholesterol, HDL cholesterol, triglyceride, glucose and insulin). RESULTS: Graded increases in BMI of Japanese and Mongolians were positively associated with body fat percent, waist circumference, hip circumference and waist/hip ratio. The Japanese were categorized as 22% overweight, 22% obese I, 3% obese II; the Mongolians rated as 18% overweight, 34% obese I, 19% obese II, based on the WPRO BMI criteria. The Mongolians had a higher prevalence of obesity and a higher body fat percent, but a lesser gradation of dyslipidemia, than did the BMI-matched Japanese groups. Overweight Japanese (BMI 23.0-24.9), in comparison to normal Japanese (BMI 18.5-22.9), had significant differences in systolic blood pressure, HDL-cholesterol and triglyceride in men, and in systolic and diastolic blood pressure, HDL cholesterol, triglyceride, insulin and Homoeostasis model assessment-insulin resistance in women. In contrast, the Mongolians showed no significant differences in metabolic parameters between overweight and normal subjects, except for diastolic blood pressure. CONCLUSION: Since the relationship between abdominal fat mass and BMI is ethnic-specific, a universal BMI cutoff point is inappropriate for Asian populations such as the Japanese and Mongolians. The present investigation suggests that, while the WPRO criteria are suitable for Japanese, the WHO criteria are more appropriate for Mongolians. PMID- 14557833 TI - Genetic basis of congenital generalized lipodystrophy. AB - Congenital generalized lipodystrophy (CGL) is an autosomal recessive disorder characterized by extreme lack of body fat and severe insulin resistance since birth. Recently, mutations have been reported in 1-acylglycerol-3-phosphate-O acyltransferase 2 (AGPAT2) and Berardinelli-Seip congenital lipodystrophy 2 (BSCL2 or Seipin) genes in affected subjects from pedigrees linked to chromosomes 9q34 and 11q13, respectively. The AGPAT2 catalyses the acylation of the lysophosphatidic acid at the sn-2 position to form phosphatidic acid, a key intermediate in the biosynthesis of triacylglycerol and glycerophospholipids. High expression of AGPAT2 mRNA in adipose tissue compared to other isoforms suggests that the mutations might affect the adipose tissue the most. The function of BSCL2 remains unknown. Several CGL pedigrees reveal no mutation in either of the above genes and are not linked to these loci, suggesting additional genetic loci for CGL. Thus, several distinct mechanisms can lead to extreme lack of adipose tissue in humans and cause CGL. PMID- 14557834 TI - [The inappropriate usage and adverse effects of drugs in clinical practice]. PMID- 14557835 TI - [Which device should be used to treat wheezing attacks in children? A nebulizer or a metered-dose inhaler?]. PMID- 14557836 TI - [Pediatric hematopoietic cell transplantations: growing pains]. PMID- 14557837 TI - [Maternal allegations for weaning: qualitative study]. AB - OBJECTIVE: To study the reasons for weaning given by women receiving care at a Baby-Friendly Maternity in Teresina, state of Piaui, Brazil. METHODS: The methodological principles of qualitative research were applied, supported by the social representation theory. Data were collected from 24 women who were in the process of weaning their babies before the 4th month of life. RESULTS: The decision-making process that leads women to wean their babies is complex and guilt-ridden. The following reasons for weaning were mentioned: having weak or little milk; puerperal problems affecting the breasts; lack of experience; disparity between the needs of the mother and the needs of the baby; external factors; work; ambiguity between wish/capacity to breastfeed and between burden/desire. CONCLUSIONS: A sense of solitude/isolation on the part of the mother and the need for support, not only from health services, but also from other segments of society, were conveyed in all interviews. The model of health care providing should be amended so as to treat breastfeeding as an act to be learned by women and protected by society. PMID- 14557838 TI - [Impact of otitis media on language acquisition in children]. AB - OBJECTIVE: To review the literature concerning the impact of otitis media on language acquisitions in children. SOURCES OF DATA: Medline. The following keywords were used for searching: otitis media, hearing, language and children. SUMMARY OF THE FINDING: The hearing loss observed in otitis media may be a result of: 1) structural disorders in the middle ear; 2) ultrastructural lesions and biochemical anomalies in the cochlea or 3) abnormalities in the central auditory pathways. About 50% of cases of secretory otitis media, for instance, are accompanied by mild fluctuating hearing loss. According to the literature, since the first three years of life are critical for language development, children affected by otitis media in this period are at risk for presenting disturbances in language acquisition, behavior and later academic achievement. CONCLUSIONS: The main consequences of otitis media and hearing loss on language acquisition are phonetic and speech articulation problems, as well as impairment of reading comprehension. PMID- 14557839 TI - [Prescription of drugs not appropriate for children in a Pediatric Intensive Care Unit]. AB - OBJECTIVE: To assess the extent of use of drugs not appropriate for children in prescriptions issued in a tertiary pediatric intensive care unit (PICU), according to FDA standards. METHODS: Observational cross-sectional study. The prescriptions issued to all patients admitted to the PICU at Hospital de Clinicas de Porto Alegre, Brazil, over a six-week period were assessed. Patients' age, sex, weight, prior disease, reason for admission to the PICU and pediatric index of mortality (PIM) were recorded, as were all drugs prescribed, their indications, presentations, doses, frequencies and means of administration. Adequacy for prescription of drugs in three pediatric age ranges was defined according to USA Food and Drug Administration (FDA) approval classification, based on the USP DI 2001 drug reference database. RESULTS: Data were obtained in the months of July and August 2002, on different days, for six consecutive weeks, based on prescriptions issued to 51 patients in 54 admissions to the PICU. Median patient age was 10.5 months; 61% of patients were male. Two thirds of patients (65%) presented prior disease. 87% of admissions were due to clinical reasons, of which 57% were respiratory complaints. A total of 747 prescription items were registered, with prevalence of 10.5% for non-approved uses and 49.5% for off label uses. No statistically significant difference was found in the distribution of prevalence of irregular prescription either by the three age ranges or by level of severity of disease at admission (according to PIM risk categories). CONCLUSIONS: The high prevalence of prescription of drugs not appropriate for children confirms, in the Brazilian context, the inadequate and inadvertent use of drugs either not approved or off-label for PICU use. This demonstrates the need to encourage further studies on the quality, efficacy and safety of drugs for pediatric use. PMID- 14557840 TI - [Metered-dose inhalers with home-made spacers versus nebulizers to treat moderate wheezing attacks in children]. AB - OBJECTIVE: To compare the effectiveness of salbutamol administration by metered dose inhaler with a home-made spacer versus jet nebulizer in children with moderate wheezing attacks. MATERIAL AND METHODS: A randomized, single-blinded trial was performed with a convenience sample of children presenting wheezing. The children were enrolled in an emergency room and randomly assigned to one of two treatment groups: home-made spacer group or nebulizer group. Clinical scores and oxygen saturation were recorded at baseline and 15 minutes after salbutamol administration. Treatment with salbutamol (100 microg/3 kg in the spacer group, and 250 microg/3 kg in the nebulizer group) was repeated at 20-minute intervals, until the child was considered to have improved significantly, with no need of any further dose, or until three doses were delivered. Treatment cost, time spent to prepare and deliver the drug, and level of parental satisfaction with the treatment were recorded. RESULTS: Fifty-four children with age between 22 days and 11.7 years were enrolled--27 in each group. Baseline and demographic characteristics were similar for both groups. The spacer was as effective as the nebulizer in terms of clinical score and oxygen saturation. The different doses (100 microg/3 kg with the spacer, and 250 microg/3 kg with the nebulizer) were shown to be clinically equivalent. Treatment cost was significantly lower in the spacer group, as was the time to prepare and deliver the drug. Parental satisfaction was similar for both inhaler devices. CONCLUSION: The home-made spacer with a metered-dose inhaler is a cost-effective alternative to a jet nebulizer in the delivery of salbutamol to children with moderate wheezing attacks. PMID- 14557841 TI - [Clinical and epidemiological analysis of bone marrow transplantation in a pediatric oncology unit]. AB - OBJECTIVE: To describe the demographics and the most important acute clinical complications in patients undergoing bone marrow transplantation in the Pediatric Oncology Unit at Hospital de Clinicas de Porto Alegre, Brazil. MATERIAL AND METHODS: A retrospective analysis was performed including 41 patients treated between August 1997 and June 2002. Twenty patients received allogeneic transplants (AG) and 21 received autologous transplants (AT). RESULTS: The mean age of AG patients was 8.9 +/- 5.4 years. Twelve patients were male. Stem cell sources were: bone marrow in 12 patients; peripheral blood in five; and unrelated cord blood in three. The diseases were acute lymphoid leukemia in seven patients; acute myeloid leukemia in four; chronic myeloid leukemia in two; myelodysplastic syndrome in two; Burkitt's lymphoma in one; severe combined immunodeficiency in one; Chediaki-Higashi syndrome in one; Fanconi anemia in one; and aplastic anemia in one. One patient developed grade II acute graft-versus-host-disease (GVHD), and three patients had grade IV GVHD. Three patients developed chronic GVHD. In all of them, the cell source was peripheral blood. Survival in this group was 70.0 +/- 10.3%. The main cause of death was GVHD in three patients and sepsis in another three. All deaths occurred before day 100. One of the patients who received unrelated cord blood is alive 3.5 years after the transplantation. In AG patients, mean age was 8.7 +/- 4.3 years. Eleven patients were male. The stem cell sources were: peripheral blood in 16; bone marrow in three; and peripheral blood + bone marrow in two. The diseases were: Wilms' tumor in five patients; Ewing's sarcoma family tumors in four; neuroblastoma in three; Hodgkin's disease in three; non-Hodgkin's lymphoma in one; rhabdomyosarcoma in two; neuroectodermic tumor of the central nervous system in two; acute myeloid leukemia in one. Survival in this group was 59.4 +/- 11.7%. Five patients died due to tumor relapse, two patients due to sepsis and one patient died in remission 20 months after bone marrow transplantation due to infection. In the whole group, the most common toxicities were vomiting, mucositis, diarrhea and abdominal pain. Infections were recorded in 58.5% of the patients. In 46.9%, at least one pathogen was isolated in the blood culture. The time required for neutrophil and platelet engraftment was correlated to the number of hematopoietic stem cell infused. CONCLUSION: The overall survival in our patients is similar to that reported in the literature. We did not find differences between AT and AG patients regarding acute toxicities and infections. PMID- 14557842 TI - [Clinical and laboratory findings in a series of cases of infective endocarditis]. AB - OBJECTIVE: To describe clinical and laboratory data of infective endocarditis (IE) in 28 children from Vitoria, state of Espirito Santo, Brazil. METHODS: We reviewed the medical records of 28 children aged 18 years and under admitted to the Infectious Diseases Unit of Nossa Senhora da Gloria Children's Hospital with a diagnosis of IE from January 1993 to December 2001. The diagnosis of IE was based on the criteria established by the Duke Endocarditis Service (Duke University, Durham, North Carolina, USA): positive blood cultures and echocardiogram (primary criteria); and fever, history of heart disease, recent heart murmur, and vascular and immunological phenomena (secondary criteria). A specific protocol was filled out by interns and medical students and revised by physicians from the hospital medical team to obtain the following data: age, sex, clinical and laboratory findings, and results of transthoracic echocardiography. In all cases, three blood samples were collected under aseptic conditions and inoculated in aerobic and anaerobic environments. The samples were then incubated at 37 degrees C and tested with the VITEC SYSTEM automatized system (BIOLAB). RESULTS: Among 28 patients, 16 were boys. Age was between 3 and 180 months (mean 70.6 +/- 59.2 months). Most children (68%) came from the metropolitan area of Vitoria, and half presented a history of cardiopathy. The most frequent clinical findings which led to suspicion of IE were: prolonged fever in all cases, new or changing regurgitant murmur (67.9%), dyspnea (57.1%), hepatomegaly (57.1%), vascular phenomena (32.2%), splenomegaly (28.6%), and Osler's nodes (7.1%). Blood cultures were positive in 16 patients (57.1%), and community-acquired S. aureus was the most frequently isolated pathogen (56.6%). Despite the treatment, fever persisted for 2.5 to 30 days (median=18.0 days). Mean leukocyte count at admission was 11,657 +/- 7,085 mm3. Transthoracic echocardiography showed endocardial lesion in all patients, evidencing vegetations in the tricuspid valve (25%), mitral valve (25%), and intraventricular communication border (28%). Thirteen (46.4%) children presented concomitant sepsis and three (10.7%) developed hospital-acquired infection. There was one death (3.6%). CONCLUSIONS: IE in our country is frequent among children younger than 2 years with congenital cardiopathy. Community-acquired S. aureus was the most frequently isolated microorganism in children with no previous cardiac lesion. Acute infective endocarditis, bacterial endocarditis, Staphylococcus aureus, transthoracic echocardiography. PMID- 14557843 TI - [Importance of clinical and laboratory profiles for the differential diagnosis of malaria and acute viral hepatitis]. AB - OBJECTIVE: To establish clinical and diagnostic findings of malaria and acute viral hepatitis in children, stressing similarities and differences, so as to enhance the sensitivity of early malaria diagnosis in childhood. METHODS: Two groups were studied, each including 30 children between 2 and 10 years of age. The patients presented either primary malaria infection or acute viral hepatitis, confirmed by thick blood film and tests for markers of viral hepatitis A and B. The patients were submitted to the following evaluations: erythrocyte, leucocyte and platelet counts, hemoglobin and hematocrit dosage, hepatic enzymes, urea, creatinine and bilirubin dosage. Clinical and laboratory findings were described for both groups and compared. Individuals with alterations on the physical exam in both groups were compared using Fisher's exact test. RESULTS: Baseline clinical findings were the same in all patients: fever, headache, digestive problems and dark urine. One half of malaria patients did not present the classical malaria signs, but all of them presented fever, differently from patients with hepatitis. In malaria patients, anemia and thrombocytopenia were significantly more frequent than in hepatitis patients. A remarkable increase of bilirubin and hepatic enzyme levels was found in hepatitis patients. CONCLUSIONS: A detailed physical examination and a thorough evaluation of non-specific laboratory tests are sufficient to allow the presumptive diagnosis of both malaria and viral hepatitis, and to reinforce the early diagnosis and treatment of malaria. PMID- 14557844 TI - [Clinical and transcutaneous oxygen saturation characteristics in hospitalized infants with acute viral bronchiolitis]. AB - OBJECTIVE: To describe the clinical characteristics of infants with acute viral bronchiolitis (AVB) and to assess the influence of oxygen desaturation time (DT) as a prognostic test to estimate the evolution of such patients. METHODS: We performed a cohort study with 111 hospitalized patients diagnosed with AVB receiving oxygen therapy through nasal prong. The outcomes were: length of admission, length of oxygen therapy and time elapsed to read 95% saturation in room air. A severity score was obtained twice a day based on clinical signs during the time when the patient required oxygen supplementation. After the supply of oxygen was interrupted, the time required for transcutaneous oxygen saturation decreased to 90% and 85%. The Chi2 test or Fisher's exact test were used to compare categorical variables. The t test or Mann-Whitney's test were used for numerical variables. Spearman's correlation was used to evaluate associations in continuous variables with asymmetric distribution. RESULTS: Most patients (61.3%) were younger than 4 months. Patients with wheezing history (45%) were analyzed separately and had similar results to those of the group with AVB (p< or =0.05). Twenty-six patients (23%) had moderate or severe malnutrition. All patients were using bronchodilators; 20% were using systemic corticosteroids; and 47%, antibiotics. The median time of oxygen therapy required for a reading of 95% oxygen saturation in room air was 83 hours (IQI 55-128). The median of length of admission was 7 days (IQI 5-10.5). Little clinical variability was observed in the period studied. No significant correlations were found between the clinical scores, DT and the outcomes. CONCLUSION: DT was not useful as an aid to assess AVB patients on oxygen therapy in this study. It is possible that this tool could have been more useful in patients with more clinical variability. PMID- 14557845 TI - [Upper airway cellular pattern in infants with acute bronchiolitis: neutrophils or eosinophils?]. AB - OBJECTIVE: To analyze the cellular pattern of nasopharyngeal secretions in infants with acute bronchiolitis (AB), focusing on the presence or absence of neutrophils and eosinophils. METHOD: Hospitalized children with AB admitted to Hospital Sao Lucas, Porto Alegre, Brazil, between May and July 2002 were recruited. Nasopharyngeal aspirates were collected during the first 48 hours after admission. Slides were stained with May Grunwald and Giemsa. Total cell count and cellular viability were obtained in all samples. RESULTS: Thirty-eight infants with AB were enrolled. The mean age was 2.2 months (interquartile range: 1.2-3.5), and 21 subjects were male. Neutrophils were the predominant cells in the nasopharyngeal aspirates (median 95%, interquartile: 94-97). No eosinophils were found in the samples studied. CONCLUSION: Our results suggest that eosinophils do not play a significant role in the pathophysiogenesis of AB. Infants with AB present a specific inflammatory response to viral infections, which is distinct from the immune response observed in asthma. PMID- 14557846 TI - [Child feeding and diarrhea morbidity]. AB - OBJECTIVE: To evaluate the protective role of breastfeeding against diarrhea in children younger than one year of age in the city of Feira de Santana, Brazil, in 2001. METHODS: A cross-sectional study was carried out. Questionnaires were applied to mothers by 104 university students on the national vaccination day in 44 health units (71.0%) selected by simple stratification. 2,319 children were evaluated (24.3% of the estimated population). The prevalence ratio was calculated and considered significant if p< or =0.05 with a 95% confidence interval. RESULTS: Diarrhea occurred in 11.6% of the sample, with greater frequency after the sixth month (63.3%). The chance for presenting diarrhea was 64% higher in children younger than six months who were not breastfed vs. breastfed children (p<0.02). When compared to the children who were exclusively breastfed, the chance for presenting diarrhea increased to 82% in children who were not breastfed (95% CI 1.11-3.01). CONCLUSIONS: Breastfeeding and exclusive breastfeeding were a protective factor against diarrhea in the first six months of life. PMID- 14557847 TI - [Assessment of mid-upper arm circumference as a method for obesity screening in preschool children]. AB - OBJECTIVE: To evaluate if mid-upper arm circumference (MUAC) can be used for obesity screening in preschool children. METHODS: 1,090 children aged from 12 to 59.99 months were studied. Personal information, weight, height and MUAC were obtained. Sensitivity and specificity to detect obese children were calculated for MUAC to age and MUAC to height z scores. Obesity was defined as weight to height z score >2. RESULTS: 6.6% of the children had a weight to height z score >2. The best association between sensitivity (76.5%) and specificity (77.9%) to detect obese children for MUAC to age was obtained with a z score of 0.7. In terms of MUAC to height, the best association between sensitivity (79.4%) and specificity (77.6%) to detect obese children was obtained with a 0.6 score. CONCLUSIONS: MUAC to height z score was not advantageous for obesity screening when compared to MUAC to age z score. MUAC to age z score seems to be an adequate alternative method for obesity screening in preschool children. PMID- 14557848 TI - [Smoking during early adolescence. A study in schoolchildren from Argentina]. AB - OBJECTIVE: To investigate the prevalence of smoking among pre-teenagers (11 and 12 years of age) and to identify risk factors for smoking. METHOD: A cross sectional study was carried out with 2,386 schoolchildren from 19 schools in Argentina. A structured, self-completed and anonymous questionnaire was employed. RESULTS: The prevalence of smoking was 15.1%, ranging from 7.9% in the 7th grade to 23.9% in the 9th grade. The mean age at the start of smoking was 11.7 years. Peer and family pressure were statistically significant risk factors for initiating the habit. CONCLUSIONS: These findings support the recommendation to begin smoking prevention programs before early adolescence. PMID- 14557849 TI - [Chronic recurrent multifocal osteomyelitis of the mandible: report of three cases]. AB - OBJECTIVE: To report three cases of chronic recurrent multifocal osteomyelitis of the mandible, an inflammatory disease affecting one or more bones with absence of isolated microorganisms in affected areas. DESCRIPTION: The first case is a 13 year-old female presenting with pain and fever after dental treatment. The patient received antibiotic treatment for osteomyelitis, but developed progressive enlargement of the mandible and palmoplantar pustulosis. Bone scintigraphy showed intense and diffuse uptake in the mandible. The swelling decreased after indomethacin and hyperbaric oxygen therapy. Case 2 is a 9 year old female patient with recurrent pain and edema of the right mandible for three years. The diagnosis of osteomyelitis was established and amoxicillin introduced. After three months, tomography showed diffuse mandible osteolysis. Indomethacin and hyperbaric oxygen therapy were introduced, however the patient presented a relapse and was treated with prednisone, rofecoxib and methotrexate. Patient 3, a 10 year-old male, had palmoplantar pustulosis and recurrent enlargement of the mandible. Tomography showed diffuse mandible osteolysis and scintigraphy revealed intense and diffuse uptake in the mandible. The patient was treated with prednisone. Rofecoxib was replaced after two relapses. COMMENTS: Chronic recurrent multifocal osteomyelitis of the mandible is often associated with prolonged pain periods and periods of activity and remission of the inflammatory process. Its recognition is important to prevent the patient from being submitted to prolonged antibiotic therapy and unnecessary invasive procedures. PMID- 14557850 TI - [Bartter' syndrome: five cases with different clinical expression]. PMID- 14557851 TI - [Asthma in Brazilian school-age children: a public health matter?]. PMID- 14557852 TI - [The contribution of short-term video-EEG for the differential diagnosis of seizures in childhood]. PMID- 14557853 TI - Associations of polymorphisms in the vitamin D receptor gene (BsmI and FokI) with bone mineral density in postmenopausal women in Malta. AB - Previous studies have suggested that variations in the vitamin D receptor (VDR) gene are related to bone mineral density (BMD). In this study, the T-->C transition in the start codon and the G-->A polymorphism at the 3' end of the VDR gene, identified by endonucleases FokI and BsmI, respectively, were analysed and correlated with BMD in postmenopausal Maltese women ( n=104). Genotype frequencies observed for the VDR start codon polymorphism (SCP) were CC: 60.4%; CT: 30.7% and TT: 8.9%, while those observed for the 3' in this study were GG: 16.4%; GA: 51.9%; AA: 31.7%. In postmenopausal women, both lumbar and femoral BMD were observed to be highest in CC homozygotes for the FokI genotype and in GG homozygotes for the BsmI genotype, although in both groups the difference between the genotypes was not statistically significant, even after adjusting BMD for age, BMI and years since menopause. No evidence of linkage disequilibrium between the two alleles was observed. PMID- 14557854 TI - Impact of postural deformities and spinal mobility on quality of life in postmenopausal osteoporosis. AB - The objective of the study was to evaluate the impact of postural deformities and spinal mobility on quality of life (QOL) in patients with spinal osteoporosis. A total of 157 postmenopausal women aged over 60 years with osteoporosis were divided into five groups according to their postural deformities: round back (RB, n=41), hollow round back (HRB, n=33), whole kyphosis (WK, n=40), lower acute kyphosis (LAK, n=18), and normal posture (NP, n=25). QOL was evaluated using the Japanese Osteoporosis QOL Questionnaire (JOQOL) proposed by the Japanese Society for Bone and Mineral Research. This questionnaire contains six domains, with higher scores indicating higher levels of QOL. The number of vertebral fractures, thoracic kyphosis and lumbar lordosis angles, and spinal range of motion (ROM) during maximum flexion and extension were also measured with radiographs. Total QOL scores in RB, HRB, WK, and LAK groups were significantly lower than those in the NP group, and those in WK group were even lower compared with the other groups ( P<0.05). All the groups with postural deformities, but not the NP group, showed significant positive correlations between total QOL score and spinal ROM (0.521STAT3 signaling in neuroendocrine function: an integrative perspective. AB - The hormone leptin is secreted by adipose tissue in proportion to fat mass to signal the repletion of body energy stores to the neuroendocrine system. Leptin acts on neurons in the hypothalamus and elsewhere in the brain to decrease appetite and regulate the activity of the thyroid, adrenal, growth, gonadal, and lactational axes. Conversely, absence of leptin signaling initiates the neuroendocrine starvation response. Leptin mediates these effects by activating the long form (LRb) of its receptor. One LRb signal, STAT3, has recently been shown to play a critical role in the regulation of body weight and some elements of neuroendocrine function (thyroid, adrenal, lactation), although the participation of STAT3 in the gonadal and growth axes is negligible. We discuss these findings in the context of the hypothalamic neuroendocrine system as it is presently understood. PMID- 14557864 TI - Carbon dioxide effects on stomatal responses to the environment and water use by crops under field conditions. AB - Reductions in leaf stomatal conductance with rising atmospheric carbon dioxide concentration ([CO2]) could reduce water use by vegetation and potentially alter climate. Crop plants have among the largest reductions in stomatal conductance at elevated [CO2]. The relative reduction in stomatal conductance caused by a given increase in [CO2] is often not constant within a day nor between days, but may vary considerably with light, temperature and humidity. Species also differ in response, with a doubling of [CO2] reducing mean midday conductances by <15% in some crop species to >50% in others. Elevated [CO2] increases leaf area index throughout the growing season in some species. Simulations, and measurements in free air carbon dioxide enrichment systems both indicate that the relatively large reductions in stomatal conductance in crops would translate into reductions of <10% in evapotranspiration, partly because of increases in temperature and decreases in humidity in the air around crop leaves. The reduction in evapotranspiration in crops is similar to that in other types of vegetation which have smaller relative reductions in stomatal conductance, because of the poorer aerodynamic coupling of the canopy to the atmosphere in crops. The small decreases in evapotranspiration at elevated [CO2] may themselves be important to crop production in dry environments, but changes in climate and microclimate caused by reduced stomatal conductance could also be important to crop production. PMID- 14557865 TI - Temporal patterns in seedling establishment on pocket gopher disturbances. AB - Disturbances often facilitate seedling establishment, and can change the species composition of a community by increasing recruitment of disturbance-adapted species. To understand the effects of pocket gopher disturbances on alpine seedling dynamics, we examined the gopher disturbances' effects on seedling emergence and survival on gopher disturbances 0 to 5 years old. In contrast to results from most other ecosystems, these recently created gopher mounds had lower seedling emergence and survival rates than undisturbed areas. A lack of correlation between species' abundances on gopher mounds and undisturbed sites in one of the two communities studied suggested that a suite of disturbance-adapted species recruited onto the mounds. To explain low seedling emergence on recent gopher mounds, we quantified gopher mound seed banks and studied recruitment in a site with mounds that ranged from 0 to >20 years old. Seed numbers in first-year gopher mound soils were extremely low relative to undisturbed soils, and this pattern was mirrored in seedling establishment patterns over the long term. Gopher disturbance depressed seedling emergence density for the first 5 years. Subsequently, emergence density increased until at least 20 years following the disturbance. Emergence on disturbances more than 20 years old was higher than on undisturbed sites. Therefore, gopher disturbances probably facilitate seedling establishment in alpine dry and moist meadow; however, this process takes place over decades. PMID- 14557866 TI - Earlier flowering between 1962 and 2002 in agricultural varieties of white clover. AB - Date of first flowering was recorded for 40 years in the same agricultural varieties of white clover (Trifolium repens L.). Standard procedures, experimental conditions and germplasm allowed the unmediated effects of climatic trends to be observed. Flowering dates of the same varieties at the same site have become significantly earlier over this period, with an advance of first flowering of approximately 7.5 days per decade since 1978. Annual maximum, minimum and soil temperatures at the site increased during the 40-year period. First flowering dates (FFDs) were significantly negatively correlated with minimum and maximum temperatures during February and March and soil temperatures between January and April. Maximum, minimum and soil temperatures increased between 1962 and 2002 for these months. No significant correlation was seen between FFD and soil temperatures at other times of the year including the period of vernalisation during winter. The occurrence of ground frost in February was significantly correlated with later flowering. Total annual rainfall and the number of wet days in a year increased between 1962 and 2002. However, only rainfall in February had a significant impact on FFD. PMID- 14557867 TI - Design and analysis of multiple choice feeding preference data. AB - Traditional analyses of feeding experiments that test consumer preference for an array of foods suffer from several defects. We have modified the experimental design to incorporate into a multivariate analysis the variance due to autogenic change in control replicates. Our design allows the multiple foods to be physically paired with their control counterparts. This physical proximity of the multiple food choices in control/experimental pairs ensures that the variance attributable to external environmental factors jointly affects all combinations within each replicate. Our variance term, therefore, is not a contrived estimate as is the case for the random pairing strategy proposed by previous studies. The statistical analysis then proceeds using standard multivariate statistical tests. We conducted a multiple choice feeding experiment using our experimental design and utilized a Monte Carlo analysis to compare our results with those obtained from an experimental design that employed the random pairing strategy. Our experimental design allowed detection of moderate differences among feeding means when the random design did not. PMID- 14557868 TI - Response of net ecosystem gas exchange to a simulated precipitation pulse in a semi-arid grassland: the role of native versus non-native grasses and soil texture. AB - Physiological activity and structural dynamics in arid and semi-arid ecosystems are driven by discrete inputs or "pulses" of growing season precipitation. Here we describe the short-term dynamics of ecosystem physiology in experimental stands of native (Heteropogon contortus) and invasive (Eragrostis lehmanniana) grasses to an irrigation pulse across two geomorphic surfaces with distinctly different soils: a Pleistocene-aged surface with high clay content in a strongly horizonated soil, and a Holocene-aged surface with low clay content in homogenously structured soils. We evaluated whole-ecosystem and leaf-level CO2 and H2O exchange, soil CO2 efflux, along with plant and soil water status to understand potential constraints on whole-ecosystem carbon exchange during the initiation of the summer monsoon season. Prior to the irrigation pulse, both invasive and native grasses had less negative pre-dawn water potentials (Psipd), greater leaf photosynthetic rates (Anet) and stomatal conductance (gs), and greater rates of net ecosystem carbon exchange (NEE) on the Pleistocene surface than on the Holocene. Twenty-four hours following the experimental application of a 39 mm irrigation pulse, soil CO2 efflux increased leading to all plots losing CO2 to the atmosphere over the course of a day. Invasive species stands had greater evapotranspiration rates (ET) immediately following the precipitation pulse than did native stands, while maximum instantaneous NEE increased for both species and surfaces at roughly the same rate. The differential ET patterns through time were correlated with an earlier decline in NEE in the invasive species as compared to the native species plots. Plots with invasive species accumulated between 5% and 33% of the carbon that plots with the native species accumulated over the 15-day pulse period. Taken together, these results indicate that system CO2 efflux (both the physical displacement of soil CO2 by water along with plant and microbial respiration) strongly controls whole-ecosystem carbon exchange during precipitation pulses. Since CO2 and H2O loss to the atmosphere was partially driven by species effects on soil microclimate, understanding the mechanistic relationships between the soil characteristics, plant ecophysiological responses, and canopy structural dynamics will be important for understanding the effects of shifting precipitation and vegetation patterns in semi-arid environments. PMID- 14557869 TI - Ultrastructure of pigment-dispersing hormone-immunoreactive neurons in a three dimensional model of the accessory medulla of the cockroach Leucophaea maderae. AB - Locomotor activity rhythms of the cockroach Leucophaea maderae are orchestrated by two bilaterally symmetric, mutually coupled, circadian pacemakers. They lie in the optic lobes of the brain and are confined to the accessory medulla (AMe), ventro-medially to the medulla. The AMe is innervated by approximately 12 pigment dispersing hormone (PDH)-immunoreactive anterior medulla neurons (PDHMe), which are circadian pacemaker candidates in the fruitfly and the cockroach. We have developed a three-dimensional computer model of the AMe and associated structures as a framework for neuroanatomical studies. Our greatly improved understanding of this structure in space has allowed us further to subdivide the anterior PDHMe into three subgroups, i.e., large, medium-sized, and small anterior PDHMe. The synaptic connections of two of these subgroups have been examined within subcompartments of the AMe by light and electron microscopy. The large, intensely staining, anterior PDHMe contain medium-sized dense-core vesicles and form input and output synapses with profiles densely filled with clear vesicles primarily in the anterior and shell neuropil of the AMe. The medium-sized anterior PDHMe contain large dense-core vesicles and constitute input and output synapses either with profiles being densely filled with clear vesicles, or with profiles containing granular dense-core vesicles. The small, weakly staining anterior PDHMe belong to a morphological group different from the large and medium-sized PDHMe and cannot be further identified at the electron-microscopic level because of their weak PDH immunoreactivity. PMID- 14557870 TI - Loss of purinergic P2X receptor innervation in human detrusor and subepithelium from adults with sensory urgency. AB - Purinergic P2X receptors associated with the parasympathetic nerves supplying human bladder smooth muscle (detrusor) are implicated in control of detrusor contractility. The relative abundance of all seven subtypes colocalised with synaptic vesicles on parasympathetic nerves was examined in specimens from normal adult bladder and in adults with the urodynamics findings of sensory urgency (SU) to determine how receptor distribution varied in patients with a small bladder capacity. Alteration in control of detrusor innervation was examined with P2X subtype-specific antibodies and an antibody (SV2) against synaptic vesicles, using immunofluorescence and confocal microscopy. Detrusor samples were taken from: controls, at cystectomy for cancer or cystoscopic biopsy for haematuria (n=22, age 33-88 years) and adults with sensory urgency at cystoscopy/cystodistension (n=11, age 37-70 years). Normal adult specimens contained detrusor muscle innervated by parasympathetic nerves possessing large varicosities (1.2 microm) distributed along their length. These mostly all showed colocalised patches of presynaptic P2X(1,2,3,5) subtypes while presynaptic subtypes P2X(4,6,7) were present in only 6-18% of varicosities. Detrusor nerve varicosities from SU patients revealed general loss of all presynaptic P2X subtypes with the proportion containing receptors reducing to only 0.5-5% depending on P2X subtype. The same loss was recorded from the sensory nerves in the surrounding lamina propria. This specific loss of P2X receptors may impair control of detrusor distension and contribute to the pathophysiology of sensory urgency. PMID- 14557871 TI - Relief effect of vitamin A on the decreased motility of sperm and the increased incidence of malformed sperm in mice exposed neonatally to bisphenol A. AB - Administration of 50 microg of bisphenol A (BPA) for the first 5 days after birth resulted in a decrease in the percentage of moving sperm, and an increase in the incidence of malformed sperm, in the epididymides of mice at 10 weeks of age, although no marked changes were found in the testicular histology between BPA treated and vehicle-treated control mice. The deteriorating effects of 50 microg of BPA were ameliorated by the concurrent administration of 100 IU of retinol acetate (RA). Neonatal treatment with 0.5 microg of BPA for 5 days resulted in an increase in the incidence of malformed sperm, whereas the BPA effect became more severe in mice nursed by mothers fed a vitamin A-deficient (VAD) diet only a few days before and after parturition. On the other hand, neonatal treatment with 20 microg of estrogen for the first 5 days after birth resulted in an increase in the number of estrogen receptor alpha (ERalpha)-positive cells in the epithelium of the vas deferens, whereas only a few epithelial cells showed weak ERalpha positive signals in the vehicle-treated control mice at 18 days after birth. This increase, however, was suppressed by the concurrent administration of RA. Although five daily treatments with 50 microg BPA led to no significant increase in the number of ERalpha-positive cells, it may have been due to the weak estrogenic activity of BPA, as discussed. These findings clearly showed that in mice, neonatal exposure to a relatively large dose of BPA causes damage to the motility and morphology of sperm, but the BPA effect is, to some extent, inhibited by a supplement of VA, and enhanced under a VAD condition. PMID- 14557872 TI - Evidence for substantial effect modification by gender in a large-scale genetic association study of the metabolic syndrome among coronary heart disease patients. AB - Major genetic determinants of the metabolic syndrome - a clustering of abdominal obesity, high triglycerides, low HDL cholesterol, high blood pressure and high fasting glucose - remain elusive. We surveyed 207 single-nucleotide polymorphisms in 110 candidate genes among coronary artery disease patients, a population enriched for metabolic abnormalities. The number of abnormalities (0-5) was determined in the 214 male and 91 female patients, and the association with each polymorphism evaluated by means of ordinal regression analysis. Polymorphisms in eight genes, including LDLR, GBE1, IL1R1, TGFB1, IL6, COL5A2, SELE and LIPC, were associated with metabolic syndrome in the whole population ( P values ranged from 0.047 to 0.008). Variants in seven additional genes showed significant gene by gender interaction. Among these, separate analyses in men and women revealed a strong association with a silent polymorphism in the low-density lipoprotein receptor-related protein gene, LRPAP1, among females ( P=0.0003), but not males ( P=0.292). Other genes associated only in females included THBS1, ACAT2, ITGB3, F2 and SELP ( P values ranging from 0.032 to 0.002). Only one gene ( PRCP) was significantly associated in men alone ( P=0.039). Our results propose several new candidate genes for the metabolic syndrome and suggest that the genetic basis of this syndrome may be strongly modified by gender. PMID- 14557873 TI - Angiostrongylus vasorum (Baillet, 1866) Kamensky, 1905: emergence of third-stage larvae from infected Biomphalaria glabrata snails. AB - Biomphalaria glabrata snails were experimentally infected with Angiostrongylus vasorum first-stage larvae and divided into four groups of 30 snails. To assess the shedding of third-stage larvae (L3), the snails were maintained under different stimuli: group 1 60 W light bulb for 24 h, group 2 37 degrees C water bath for 24 h, group 3 room temperature (23-25 degrees C) for 24 h, Group 4 room temperature (23-25 degrees C) for up to 15 days. After 24 h, a total of 512 A. vasorum L3, alive and active, were released by snails from group 1, while 2,446 L3 were released from group 2 and five L3 from group 3. After 15 days, snails from group 4 released a total of 44 L3. To evaluate the infectivity of A. vasorum L3, two mongrel dogs were successfully infected with L3 released by snails from groups 1 and 2, confirming that the infection of dogs with A. vasorum L3 was possible, independently of ingestion of the mollusk intermediate host. The results shown in these experiments suggest that angiostrongylosis could be directly transmitted to the definitive hosts, with implications for the parasite's life cycle. PMID- 14557874 TI - Evidence of horizontal transmission of feline leukemia virus by the cat flea ( Ctenocephalides felis). AB - The feline leukemia virus (FeLV) is a naturally occurring and widespread retrovirus among domestic cats. The virus is mainly transmitted horizontally through saliva, blood and other body fluids by close contact between cats. Vectors other than cats, e.g. blood-sucking parasites, have not been reported. This study tested the vector potential of the cat flea ( Ctenocephalides felis) for FeLV. In a first feeding, fleas were fed for 24 h with blood from a FeLV infected cat with persistent viremia. FeLV could be detected in the fleas, as well as in their feces. Fleas were then divided in two populations and fed in a second feeding for 5 h or 24 h with non-infected non-viremic blood. FeLV was again detected in the fleas and their feces. In addition, the two resulting blood samples of the second feeding were subsequently tested for FeLV and both samples were positive for FeLV RNA. The cat flea transmitted the FeLV from one blood sample to another. In a third feeding, the same populations of fleas were fed again with non-infected blood for 5 h or 24 h. This time FeLV was not detected in the fleas, or in the feces or blood samples. Results show that cat fleas are potential vectors for FeLV RNA in vitro and probably also in vivo. PMID- 14557875 TI - Up-regulated humoral immune response in the soft tick, Ornithodoros moubata (Acari: Argasidae). AB - Ticks have an efficient defense system for preventing microbial infection. The antimicrobial peptide defensin is one effective molecule in this system. Here we investigated immune competence and the involvement of defensin in the humoral defense of the soft tick, Ornithodoros moubata. Semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) revealed that gene expression of all four defensin isoforms was up-regulated by bacteria or bacterial components. Defensin gene up-regulation by hemocoelic inoculation of bacteria involves the midgut and granulocytes. In immunodetection analysis, immunization by bacterial injection increases the relative concentration of defensin-like material in the hemolymph plasma. Furthermore, elevated antibacterial activity against Gram positive bacteria but not against Gram-negative bacteria was observed after immunization by a liquid growth inhibition assay. Therefore, enhanced anti-Gram positive bacterial activity appears to be partially dependent on the release of defensin into the hemolymph. These findings demonstrate that defensin plays an important role in the up-regulated humoral response of O. moubata. PMID- 14557876 TI - Biochemical analysis of encapsulated and non-encapsulated species of Trichinella (Nematoda, Trichinellidae) from cold- and warm-blooded animals reveals a high genetic divergence in the genus. AB - Multilocus enzyme electrophoresis was used to analyse genetic variation in the genus Trichinella. Twenty-eight isolates belonging to eight species and six genotypes were analysed for 12 enzyme systems, producing 19 different phenotypes. According to Jaccard's similarity index, the isolates clustered into two main groups, specifically, encapsulated species/genotypes and non-encapsulated species/genotypes. Furthermore, the non-encapsulated species clustered into two other groups: the species infecting mammals and birds ( Trichinella pseudospiralis) and those infecting mammals and reptiles ( Trichinella papuaeand Trichinella zimbabwensis). The encapsulated species/genotypes, which only infect mammals, clustered into four main groups: the cosmopolitan species Trichinella spiralis, the species/genotypes of the temperate regions ( Trichinella britovi, Trichinella murrelli, Trichinella T8, and Trichinella T9), the species/genotype of the arctic region ( Trichinella nativa and Trichinella T6), and the equatorial species Trichinella nelsoni. These results are consistent with biological, epidemiological, and molecular data, which show a high genetic divergence in this genus. PMID- 14557877 TI - Effect of native Xenorhabdus on the fitness of their Steinernema hosts: contrasting types of interaction. AB - Steinernema species are entomopathogenic nematodes. They are symbiotically associated with Enterobacteriaceae of the genus Xenorhabdus. These nematode bacteria symbioses are extremely diversified and constitute an important new model in ecology and evolution to investigate symbioses between microbes and invertebrates. However, no study has so far adequately evaluated either the outcome of the interactions or the obligate nature of interactions in different Steinernema species in the same way. Studying three different species of Steinernema, we showed that symbiotic nematodes are always fitter than aposymbiotic ones. Nevertheless, we revealed contrasting types of interaction in terms of outcome and obligate nature of the interaction. Bacterial analyses showed that nematode species differed dramatically in the number of symbiotic Xenorhabdus they carried. We suggested that when the interaction appeared more facultative for a nematode species, the nematodes carried fewer Xenorhabdus cells than strongly dependent worm species. Thus, the symbiont transmission appeared to become more efficient as the relationship between the nematode and the bacteria became tighter. PMID- 14557880 TI - Interleukin-3-producing CD4+ T-cells support eosinophil survival in a patient with transient hypereosinophilia. PMID- 14557878 TI - Epidemiological aspects of cancer screening in Germany. AB - PURPOSE: Since 1971, a statutory early detection programme has operated in Germany which comprises health-insurance-paid annual examinations of the breast, cervix, prostate, rectum, and the skin. Since the programme is conceptualised as opportunistic screening, the attendance rates have been low and only reached about 50% among females and 13% among males by the end of the 1990s. Based on these figures and present knowledge on the efficacy of screening modalities, we assessed past benefits and the future potential of cancer screening in Germany. METHODS: We used published data on the efficacy of screening procedures and German attendance rates, and internationally available data on incidence and mortality in Germany and, for cervical cancer, in other countries. Incidence and mortality rates have been standardised to the world standard, and screening benefit has been given as the population preventable fraction given in percentage. RESULTS: The past benefits of the statutory early detection programme ranged around 2.0-6.5%. Since the upper limit was due to generous assumptions regarding efficacy or inclusion of treatment effects, the true value might be closer to the estimates of the effect of cervical cancer screening (2.0-4.7%). The achievable future benefit of exploiting the theoretical potential of more exhaustive screening could provide a further mortality reduction of about 3.4% (50% compliance) or 4.7% (70% compliance). CONCLUSIONS: Screening partially requires an expensive medical infrastructure and is not without risks for the participants. The overall benefit is critically dependent upon the quality of the programme and its in-time control. Any benefit may be annulled by poor quality while costs are overflowing. Well-organised high-quality screening may be a sound basis for cancer control. To preserve or increase the impact of screening and control its expenses: (a) further research efforts are needed towards new or better targeted screening tools or modalities; (b) the efficacy of new modalities has to be evaluated carefully in advance; (c) the programme has to be reconceptualised as organised screening; (d) in-time quality control based on the collection of the basic performance data must be an intrinsic part of the programme. PMID- 14557879 TI - Expression of Cdx2 and the phenotype of advanced gastric cancers: relationship with prognosis. AB - PURPOSE: The clinicopathologic significance of the phenotype and Cdx2 expression has hitherto remained unclear in gastric cancers. In the present study, we therefore examined the correlation between prognosis, phenotype, and Cdx2 expression in advanced cases. METHODS: We evaluated 177 advanced gastric carcinomas histologically and phenotypically. The expression of Cdx2 was also assessed by immunohistochemistry. RESULTS: The lesions were phenotypically divided into 32 gastric (G type), 36 gastric and intestinal mixed (GI type), 53 intestinal (I type), and 56 null (N type) types, independent of the histological classification. Cdx2 nuclear staining demonstrated a close relation to intestinal phenotypic expression, not with the histological classification. Kaplan-Meier analysis of Cdx2 expression and the phenotype showed that the Cdx2 positive groups had a significantly better outcome than the negative ones (P=0.0013), and the patients with GI type cancers had significant better survival than those with N type (P=0.0052). CONCLUSIONS: Our results suggest that Cdx2 is a useful prognostic marker. In addition, advanced gastric cancers with both intestinal and gastric phenotypic expression have a relatively good prognosis. Combined evaluation of gastric and intestinal epithelial cell markers, including Cdx2, is clinically useful to predict outcome in patients with advanced neoplasm of the stomach. PMID- 14557881 TI - The maize disorganized aleurone layer 1 and 2 ( dil1, dil2) mutants lack control of the mitotic division plane in the aleurone layer of developing endosperm. AB - The maize ( Zea mays L.) endosperm consists of an epidermal like layer of isodiametric aleurone cells surrounding a central body of starchy endosperm cells. In disorgal1 ( dil1) and disorgal2 ( dil2) mutants the control of the mitotic division plane is relaxed or missing, resulting in mature grains with disorganized aleurone layers. In addition to orientation of the division plane, both the shape and size of the aleurone cells are affected, and often more than one layer of aleurone cells is present. Homozygous dil1 and dil2 grains are shrunken due to reduced accumulation of starchy endosperm and premature developmental arrest of the embryo, and mature mutant grains germinate at a very low rate and fail to develop into plants. However, homozygous mutant plants can be obtained through embryo rescue, revealing that both mutants have an irregular leaf epidermis as well as roots with a strongly reduced number of root hairs and aberrant root hair morphology. Our results suggest the presence of common regulatory mechanisms for the control of cell division orientation in the aleurone and plant epidermis. PMID- 14557883 TI - How long do we need teaching in the operating room? The true costs of achieving surgical routine. AB - BACKGROUND: Our aim was to quantify the incremental costs of longer operating times of residents and less-experienced junior consultants when compared with senior consultants on the basis of two surgical routine procedures. METHODS: We prospectively assessed 246 patients who underwent laparoscopic cholecystectomy and 216 patients who underwent open inguinal hernia repair. Operating times, complication rates and overall costs for these patients were recorded and linked to the attending surgeons. RESULTS: Most importantly, operating times significantly depend on the surgeon (P<0.001) and on proper supervision of junior surgeons (P<0.001 to P=0.003). When compared with those of senior surgeons, incremental costs for the hospital provider were Euro 200 and Euro 54 per laparoscopic cholecystectomy and Euro 153 and Euro 106 per open hernia repair when carried out by junior consultants and residents, respectively. Overall incremental costs per year for these procedures were Euro 8,370 for residents and Euro 22,922 for junior consultants. CONCLUSION: Owing to longer operating times for junior consultants the costs of achieving surgical routine are considerably higher than previously estimated. These higher costs derive from junior consultants performing operations without proper supervision from senior consultants. We conclude that prolonged supervision in the operating room is highly cost-effective regardless of higher costs for personal resources per operating-minute. PMID- 14557882 TI - Acetylcholine induces Ca2+ oscillations via m3/m4 muscarinic receptors in the mouse oocyte. AB - Changes in intracellular Ca2+ concentration are required for the activation of mammalian oocytes. They are caused mainly by Ca2+ release from the endoplasmic reticulum (ER) via Ins P3 receptors (Ins P3R). Several studies have reported that acetylcholine (ACh) is capable of triggering early activation events in mouse oocytes over-expressed with the m1 muscarinic ACh receptor (m1AChR). Here we examined which subtypes of the mAChR (m1 to m4) are involved in the generation of Ca2+ oscillations in native mouse oocytes. ACh (10 microM) elicited regular Ca2+ oscillations similar to those induced by sperm in their temporal characteristics. The Ca2+ oscillations were abolished by application with atropine, the mAChR inhibitor. Within 1 min after treatment of ACh, intracellular Fluo-3 fluorescence intensity increased from 794+/-119 to 2023+/-755 (increase to 250% of original value), indicating a strong rise of cytosolic Ca2+ concentration. 4-DAMP mustard and Tropicamide, specific antagonists of m3AChR and m4AChR, completely abolished ACh-induced Ca2+ oscillations. In the ovulated oocytes, the expression of m3/m4 AChR was clearly detected by RT-PCR analysis. Furthermore, ACh-induced Ca2+ oscillations were also abolished or decreased by PLC inhibitors (U73122 or D609) and an Ins P3-receptor antagonist (xestospongin C), confirming that ACh generates Ca2+ oscillations via the PLC-Ins P3 (PI) pathway. These results strongly suggest that m3/m4AChR is coupled to the generation of Ca2+ oscillations mainly via the PI pathway in mouse oocytes. PMID- 14557884 TI - Heart rate variability during cycloergometric exercise or judo wrestling eliciting the same heart rate level. AB - This study compared heart rate variability (HRV) in ten male judokas between two types of exercise eliciting the same near-maximal average heart rate (HR): judo wrestling vs. cycloergometric bout. Beat-to-beat RR intervals were recorded during (1) a 4-min judo randori (wrestling); (2) a 4-min cycloergometric exercise eliciting maximal oxygen consumption (VO(2MAX)). Time series were analyzed both by short term Fourier transform (STFT) and Poincare plot (PP). The main results are as follows. First, despite the fact that the same maximal HR was reached during the two exercises, the spectral energy computed from the judo recordings was significantly higher than that recorded from the cycloergometric exercise. Second, according to the PP index of rapid HRV (SD1), the high-frequency spectral energy (HF) was significantly higher during judo than cycloergometric exercise as well. Third, judo spectra show chaotic harmonics in place of the precise HF peak observed during cycloergometric exercise. Fourth, the respective parts of normalized LFn and HFn are not different between the two exercise modes, suggesting that autonomic control during severe exercise cannot depend on the type of exercise. In conclusion, this study shows that it is possible, according to the observed kind of variability from RR time series, to differentiate between two types of effort: steady-state dynamic exercise or conversely exercise made of both isometric and irregular dynamic efforts (wrestling, collective sports, and others). PMID- 14557885 TI - Acute and severe hypobaric hypoxia-induced muscle oxidative stress in mice: the role of glutathione against oxidative damage. AB - This study intended to analyze: (1) the effects of acute and severe hypoxia exposure on skeletal muscle oxidative stress and oxidative damage markers; (2) the protective role of the antioxidant glutathione against oxidative damage; and (3) the expression of heat shock protein 70 kDa (HSP70) induced by this hypoxic insult. Forty mice were divided into four groups: control + placebo (C+P), hypoxia + placebo (H+P), control + l-buthionine-[ S, R]-sulfoximine (BSO, a GSH depleting compound) (C+BSO) and hypoxia + BSO (H+BSO). Hypoxia groups were continuously exposed for 24 h to a hypobaric hypoxic environment equivalent to an altitude of 7000 m and sacrificed immediately after. Control groups were maintained at sea level during the experimental protocol. Analyzed biochemical parameters were: reduced (GSH) and oxidized (GSSG) glutathione, thiobarbituric acid reactive substances (TBARS), sulfhydryl protein groups (SH), N-acetyl-beta- d-glucosaminidase (NAG) and HSP70 protein. Hypoxia (H+P) per se, compared to C+P, induced a significant increase in %GSSG (5.68 vs. 1.14%), TBARS (436.7 vs. 227.9 nM), NAG (4.49 vs. 3.35 U/mg) and HSP70 (178.7 vs. 100%). Compared with H+P, H+BSO showed a significant decrease in total glutathione (19.30 vs. 6.13 nmol/mg) and an additional increase in %GSSG (5.68 vs. 11.33%) and in HSP70 expression (178.7 vs. 202.2%). However, no further oxidative damage was observed in H+BSO. These data suggest that acute hypoxia per se might enhance oxidative stress; however, the glutathione system seems to have a modest role in skeletal muscle protection against hypoxia-induced oxidative stress. Moreover, hypoxia and BSO treatment is a sufficient stimulus to promote HSP70 overexpression. PMID- 14557887 TI - Dysphonia: medical treatment and a medical voice hygiene advice approach. A prospective randomised pilot study. AB - For many years all patients with dysphonia referred to in the literature as resulting from non-organic (functional) voice disorders were sent to speech therapy. Medical diagnoses were not taken into account. In our earlier Cochrane review on vocal cord nodules we discovered that evidence-based research in the area of benign voice disorders with dysphonia, and with or without slight benign swellings including nodules on the vocal cords, was lacking at that time. Therefore, a prospective randomised pilot study based on our Cochrane review has been made on dysphonic patients with non-organic (function provoked?) voice disorders as the basis for further evidence-based studies. Medical treatment was based on the scientific approach that once a micro-organic disorder caused by reflux, infection, allergy or environmental irritatants (e.g., dust or noise in the workplace) was discovered by very careful anamnesis and systematic objective routine analyses and was treated effectively, with documentation, the non-organic voice disorder disappeared, as, e.g., in the case of a diagnosis and treatment of helicobakter pylori. The reason is that the mucosal swelling/dysfunction of the vocal cords is secondary. In order to try to understand why the recommendation to all these patients for many years was only voice therapy, which the speech therapists "felt to be effective", updated voice-hygiene advice (for posture, accents of the diaphragm, intonation pattern and resonance) was given by experienced laryngologists, randomised with the updated medical diagnosis/therapy in order to elucidate what effect the training might have. No evidence-based studies in the literature document any effect. The crucial point seemed to be that doctors mostly did not examine any other diagnoses other than the "dysphonia" and did not dig down to any of the medical reasons when the vocal fold diagnosis of "non- organic disorders" was made. This should be changed in the future. This pilot study was based on a comparison of ten dysphonic patients with stroboscopic non- organic (functional) voice disorders, where a micro organic diagnosis was searched for and treated systematically in a medical regime (for infections, allergies, gastrooesophageal reflux and environmental irritants such as dust, noise, etc.) versus ten dysphonic patients with stroboscopically confirmed non-organic (functional) voice disorders, having only the traditional but optimal voice advice, which we can call medical voice-hygiene advice, including the use of the Accent method. A retrospective group of ten patients treated medically was included, too. A demand cannot be made that the functional group being treated by randomisation with voice advice should also be medically treated at once, the medical approach being the new one. On the other hand, it is strange that no evidence-based research was made before. All patients were measured two times with stored videostroboscopy, a quality-of-life questionnaire and phonetograms with 1-month intervals. All patient groups improved. There was no statistical improvement in favour of the medical group with the voice-related quality-of-life score, also not for the group who received voice-hygiene advice. The geometrical mean values of the phonetogram areas in decibels times semitones were better in all groups, but a statistical difference was not found between the medically treated group and the voice-hygiene advice group. The pilot study showed that both medical treatment and medical voice-hygiene advice had a positive effect on dysphonia in non-organic (functional) voice disorders. There is need of an extensive prospective randomised trial on dysphonia including vocal cord nodules to find out which treatment should be used for this group of patients. It is suggested that an eventual randomisation for microsurgical treatment or regular voice therapy should be made after a period of systematic medical diagnosis and treatment including medical voice-hygiene advice. PMID- 14557888 TI - Significance of CA 125 serum level in discrimination between benign and malignant masses in the pelvis. AB - AIM: Our aim was to confirm that preoperative CA 125 serum level can be useful for discrimination between benign and malignant masses in the pelvis. METHODS: Preoperative CA 125 serum level was analyzed retrospectively in 121 patients who had surgery because of a malignant ovarian tumor and in 91 patients with benign masses in the pelvis. The cutoff serum level CA 125 between benign and malignant masses in the pelvis was 35 and 65 IU/ml. RESULTS: Of those patients with a malignant ovarian tumor, 65.3% had menopause whereas only 31.5% of those with a benign tumor did so. The average age of the patients with a malignant tumor was 54.2 years and of those with a benign tumor 46.8 years. The preoperative CA 125 serum level was higher than 35 IU/ml in 80.2% and higher than 65 IU/ml in 72.7% of all analyzed patients with a malignant tumor, whereas it was 23.9% and 9.8% respectively in patients with a benign mass. In early stage ovarian cancer disease (borderline stage, I/II) the preoperative CA 125 serum level was higher than 35 IU/ml in 67.8% and in 52.5% higher than 65 IU/ml. In advanced stages (III/IV), it was higher than 35 and 65 IU/ml in 96.1%. After therapy the CA 125 serum level dropped below 35 IU/ml in 70.8% and after three chemotherapy courses in 78.1%. A CA 125 level less than 35 IU/ml was achieved by therapy in 84.2% patients with an early stage disease (I/II) and in 62.1% in advanced stages (III/IV). The calculated sensitivity was 80.2% and negative 74.5% (CA 125 higher than 35 IU/ml) and 72.7%, 90.2%, 90.7%, 71.6% respectively (CA 125 higher than 65 IU/ml). CONCLUSION: . Preoperative determination of CA 125 is a very useful method to discriminate between benign and malignant masses in the pelvis. PMID- 14557889 TI - True ectopic ovary: a case and review. AB - INTRODUCTION: Ectopic ovarian tissue is a rare gynecologic problem. The nomenclature and classification systems are unclear and still debated. CASE REPORT: Herein we provide a case report and review the literature to help clarify the classification system of ectopic ovarian tissue. PMID- 14557886 TI - Differential binding of platelet-derived growth factor isoforms to glycosaminoglycans. AB - The platelet-derived growth factor (PDGF) family comprises disulfide-bonded dimeric isoforms and plays a key role in the proliferation and migration of mesenchymal cells. Traditionally, it consists of homo- and heterodimers of A and B polypeptide chains that occur as long (A(L) and B(L)) or short (A(S) and B(S)) isoforms. Short isoforms lack the basic C-terminal extension that mediates binding to heparin. In the present study, we show that certain PDGF isoforms bind in a specific manner to glycosaminoglycans (GAGs). Experiments performed with wild-type and mutant Chinese hamster ovary cells deficient in the synthesis of GAGs revealed that PDGF long isoforms bind to heparan sulfate and chondroitin sulfate, while PDGF short isoforms only bind to heparan sulfate. This was confirmed by digestion of cell surface GAGs with heparitinase and chondroitinase ABC and by incubation with sodium chloride to prevent GAG sulfation. Furthermore, exogenous GAGs inhibited the binding of long isoforms to the cell membrane more efficiently than that of short isoforms. Additionally, we performed surface plasmon resonance experiments to study the inhibition of PDGF isoforms binding to low molecular weight heparin by GAGs. These experiments showed that PDGF-AA(L) and PDGF-BB(S) isoforms bound to GAGs with the highest affinity. In conclusion, PDGF activity at the cell surface may depend on the expression of various cellular GAG species. PMID- 14557890 TI - Clinical results after restorative proctocolectomy without diverting ileostomy for ulcerative colitis. AB - BACKGROUND AND AIMS: This study evaluated the postoperative complications and clinical results of restorative proctocolectomy without diverting ileostomy for ulcerative colitis. PATIENTS AND METHODS: One hundred selected patients had a hand-sewn ileal J-pouch anal anastomosis with mucosectomy using an ultrasonically activated scalpel. RESULTS: Three patients with pouch-related complications who needed diverting ileostomy. Five patients showed intestinal obstruction; two of the five needed relaparotomy and division of adhesions. The median number of bowel movements per 24 h was 6.5 (2-13) at 3 months and 5 (3-10) at 12 months. The corresponding nightly frequencies were 0 (0-5) at 3 months and 0 (0-3) at 12 months. After 3 months 82% of patients had no soiling during the daytime, and 45% were fully continent day and night. After 3 months 89% had recovered the ability to distinguish flatus from feces. CONCLUSION: Ileal pouch anal anastomosis can be performed safely without diverting ileostomy using an ultrasonically activated scalpel. The postoperative functional result was stabilized 3 months after the operation. PMID- 14557891 TI - The clear cell adenocarcinoma of the colon: case report and review of the literature. AB - BACKGROUND: Primary clear cell adenocarcinoma of the colon is a rare oncological entity. We report a case of such a tumor in the left colonic flexure of an 89 year-old patient who was treated surgically in our Department. We also review published cases which include primary colonic clear cell formations. CASE PRESENTATION: The patient sustained a segmental resection of the region [pT2 N0 (0/12) M0]. Histochemical examination indicated a primary clear cell colonic adenocarcinoma. The patient was discharged in good health but died from his disease 2 years later. RESULTS: Primary clear cell colonic adenocarcinoma is found exclusively in the left colon and generally in elderly men. From the few cases that have been published it is not possible to estimate the oncological impact of this tissue. CONCLUSION: Even in advanced age surgical resection is the mainstay of the recommended treatment. PMID- 14557892 TI - Total cystectomies in the surgical treatment of rectal cancer with prior chemoradiation: analysis of postoperative morbidity and survival. AB - BACKGROUND AND AIMS: Curative surgery for rectal cancer seldom requires urinary tract resections. The study investigated morbidity and survival following resection of rectum with total cystectomy following chemoradiation for primary rectal cancer. PATIENTS AND METHODS: 19 consecutive patients with primary nonresectable rectal cancer undergoing preoperative chemoradiation and operated on by a multidisciplinary team of surgeons. RESULTS: Morbidity was moderately low, and only five cases required surgical reintervention. No postoperative deaths were observed. Long-term survival in this group of patients compares well with the survival of patients with primarily nonresectable rectal cancer without the involvement of urinary bladder. CONCLUSION: Extended pelvic exenteration due to rectal cancer is relatively safe and in selected patients offers long-term survival and a chance of a cure. Involvement of the urinary bladder does not adversely affect outcome of rectal cancer treatment. PMID- 14557893 TI - Introduction of a urodynamic score to detect pre- and postoperative neurological deficits in children with a primary tethered cord. AB - OBJECT: An increasing number of asymptomatic children are diagnosed with occult spinal dysraphism, raising the question of their optimal management. Urodynamic study (UDS) is the most reliable method of detecting neuro-urological abnormalities in these children. The rate of postoperative retethering ranges from 10 to 20% and is not always immediately clinically significant. The aim of this prospective study was to develop a reliable method that could be used in the preoperative assessment and postoperative follow-up of children with a tethered cord syndrome (TCS). METHODS: From 1989 to 1997, 15 children underwent spinal cord untethering for TCS. Preoperatively, patients were assessed with MRI and UDS. Postoperative UDS were repeated at 6- to 12-month intervals. Four UDS parameters were identified, graded, and added to obtain a UDS score. A group of 38 children without dysraphic condition was used as control and allowed the calculation of a normal score. CONCLUSIONS: There was a statistically significant difference in the preoperative UDS scores between the control group and the study group ( p<0.001). Postoperatively, there was a statistically significant improvement ( p<0.001) in UDS scores. UDS score is a reliable tool for identifying and quantifying neuro-urological disorders in patients with TCS. Postoperatively, this score was useful in the early diagnosis of spinal cord retethering. PMID- 14557894 TI - MR imaging in the assessment of residual tumour following inadequate primary excision of soft tissue sarcomas. AB - The purpose of this retrospective study was twofold: firstly, to assess the ability of MR imaging in confirming/excluding the presence of residual tumour following inadequate primary excision of soft tissue sarcomas; and secondly, to assess the accuracy of the original radiologists report as compared with a retrospective review of the scan hard copy in confirming/excluding. A total of 111 cases were identified that fulfilled the inclusion criteria of inadequate primary surgery followed by a MR scan and subsequent wide re-excision of the surgical field. The gold standard for the assessment of the MR imaging studies was histological examination of the re-excision specimens. Histological examination revealed residual tumour in 63 (56.7%) cases. In 48 cases the residual tumour was classified macroscopic (maximum diameter >10 mm) and 15 cases microscopic (maximum diameter 0.01). Pure GGO were not observed. Lesion profusion was highly variable and correlated moderately negative with diffusion capacity (mean 88.2% (SD +/- 20.9%); r=-0.54; p<0.001) and very weak with vital capacity and FEV1 (mean values 92.2% (SD +/- 18.3%); r=-0.27; p<0.05 and 89.8% (SD +/- 17.5%); r=-0.31; p<0.01). In patients with GGO, BAL differentials tended towards neutrophilia (kappa=0.39; p=0.04; McNemar test p>0.2), but not towards lymphocytosis (kappa=0.10; p=0.23; McNemar test p>0.2). Differences in smoking history were not significant (p>0.1). The HRCT appears most appropriate for the detection and follow-up of ILD associated with RA. The PFT and BAL correlate only partially with lesion profusion or grading on HRCT, but they contribute valuable information about dynamic lung function and differential diagnoses (pneumonia, medication side effects). PMID- 14557896 TI - Docetaxel plus epirubicin is a highly active, well-tolerated, first-line chemotherapy for metastatic breast cancer: results of a large, multicentre phase II study. AB - PURPOSE: In this multicentre phase II study, the efficacy and safety profile of the combination of docetaxel and epirubicin as first-line chemotherapy for metastatic breast cancer (MBC) were evaluated. METHODS: Epirubicin (75 mg/m(2)) and docetaxel (75 mg/m(2)) were given intravenously once every 3 weeks for six cycles to 133 patients with MBC. RESULTS: The overall clinical response rate was 67% (complete and partial responses were 23% and 44%, respectively). The median time to progression was 10.8 months (95% CI 9.7-12.6) and the median overall survival was 19.5 months. Granulocyte colony-stimulating factor support was administered to 32% of patients and in 22% of cycles. Grade 3/4 neutropenia occurred in 35% of patients and febrile neutropenia in 19%. The most frequent grade 3/4 non-haematological toxicities (as percent of patients) were asthenia (6%), vomiting (5%) and nausea (5%). No patients developed congestive heart failure. CONCLUSIONS: The combination of docetaxel and epirubicin was highly active as first-line treatment for MBC and showed a manageable toxicity profile. PMID- 14557898 TI - Evidence that 99mTc-(V)-DMSA uptake is mediated by NaPi cotransporter type III in tumour cell lines. AB - In vivo studies have demonstrated that pentavalent technetium-99m dimercaptosuccinic acid [(99m)Tc-(V)-DMSA] may be a useful tumour imaging agent. Several studies have suggested that (99m)Tc-(V)-DMSA uptake may be related to the structural similarity between the (99m)Tc-(V)-DMSA core and the PO(4)(3-) anion. As phosphate ions enter cells via NaPi cotransporters, we investigated whether (99m)Tc-(V)-DMSA uptake is mediated by NaPi cotransporters. (99m)Tc-(V)-DMSA and phosphate uptake kinetics were compared in three cancer cell lines (MCF-7, G152 and MG-63) under several conditions (with and without sodium and NaPi cotransporter inhibitor and at different pH). Determination of molecular NaPi cotransporter mRNA expression was performed by reverse-transcriptase polymerase chain reaction (Rt-PCR) assay. Results obtained in the presence of NaPi inhibitor, in sodium-free medium and at alkaline pH showed that (99m)Tc-(V)-DMSA accumulation is linked to NaPi cotransporter functionality. MCF-7 and G152 exhibited the same tracer uptake, whereas MG-63 showed the highest phosphate accumulation and the lowest (99m)Tc-(V)-DMSA uptake. These results were in accordance with mRNA NaPi expression, i.e. all cell lines expressed NaPi type III but MG-63 also co-expressed NaPi type I. The total level of NaPi cotransporter was highly correlated with phosphate accumulation, while the level of type III was related to (99m)Tc-(V)-DMSA uptake. We have demonstrated that (99m)Tc-(V) DMSA uptake is specifically mediated by NaPi type III in cancer cells. PMID- 14557899 TI - "A Japanese gentleman of the Samurai tradition": Takeshi Matsunaga 1945-2003. PMID- 14557900 TI - Solitary fibrous tumor in the pelvic space. AB - A case of a solitary fibrous tumor (SFT) of the pelvic space in a 64-year-old man is reported herein. Computed tomography (CT) of the pelvis showed a large mass enhanced heterogeneously left paracentral and posterior to the bladder and intimately associated with prostate. The site of origin of the mass could not be defined on CT because margins blended with the bladder, prostate, and rectum. A tumorectomy was performed and has remained well with no evidence of recurrence during the last 3 months. The tumor was 12.5 x 9.5 x 8.3 cm in size, solid with a fibromuscular capsule, and gray-tan in color. Histologically, the neoplasms were well circumscribed and composed of short spindle cells arranged without an obvious pattern. Immunohistochemically, these cells were strongly positive for CD 34 and negative for S-100, alpha SMA, and AE1/AE3. PMID- 14557901 TI - Zonal frequency analysis of the gyral and sulcal extent of cerebral infarcts. Part III: Middle cerebral artery and watershed infarcts. AB - We tested the hypothesis that frequency analysis of the anatomic zones affected by single anterior (A), posterior (P), and middle (M) cerebral artery (CA), multivessel, and watershed infarcts will disclose specific sites (peak zones) most frequently involved by each type, sites most frequently injured by multiple different types (vulnerable zones), and overlapping sites of equal relative frequency for two or more different types of infarct (equal frequency zones). We adopted precise definitions of each vascular territory. CT and MRI studies of 50 MCA, 20 ACA-MCA, three PCA-MCA, and 30 parasagittal watershed infarcts were mapped onto a standard template. Relative infarct frequencies in each zone were analyzed within and across infarct types to identify the centers and peripheries of each, vulnerable zones, and equal frequency zones. These data were then correlated with the prior analysis of 47 ACA, PCA, dual ACA-PCA, and ACA-PCA-MCA infarcts. Zonal frequency data for MCA and watershed infarcts, the sites of peak infarct frequency, the sites of vulnerability to diverse infarcts, and the overlapping sites of equal infarct frequency are tabulated and displayed in standardized format for direct comparison of different infarcts. This method successfully displays the nature, sites, and extent of individual infarct types, illustrates the shifts in zonal frequency and lesion center that attend dual and triple infarcts, and clarifies the relationships among the diverse types of infarct. PMID- 14557902 TI - Assessment of brain perfusion with MRI: methodology and application to acute stroke. AB - We review the methodology of brain perfusion measurements with MRI and their application to acute stroke, with particular emphasis on the work awarded by the 6th Lucien Appel Prize for Neuroradiology. The application of the indicator dilution theory to the dynamic susceptibility-weighted bolus-tracking method is explained, as is the approach to obtaining quantitative measurements of cerebral blood flow (CBF) and volume (CBV). Our contribution to methodological developments, such as CBV measurement with the frequency-shifted burst sequence, development of the PRESTO sequence, comparison of different deconvolution methods and of spin- and gradient-echo sequences, and the validation of MRI measurements against positron emission tomography is summarised. The pathophysiology of brain ischaemia and the role of neuroimaging in the setting of acute stroke are reviewed, with an introduction to the concepts of ischaemic penumbra and diffusion/perfusion mismatch. Our work on the determination of absolute CBF and CBV thresholds for predicting the area of infarct growth, identification of the best perfusion parameters (relative or absolute) for predicting the area of infarct growth and the role of MR angiography is also summarised. We conclude that MRI is a very powerful way to assess brain perfusion and that its use might help in selecting patients who will benefit most from treatment such as thrombolysis. PMID- 14557903 TI - Giant spinal perimedullary fistula in hereditary haemorrhagic telangiectasia: diagnosis, endovascular treatment and review of the literature. AB - Hereditary hemorrhagic telangiectasia (HHT, Rendu-Osler-Weber disease) is a vascular disorder with dominant autosomal transmission characterised usually by multiple mucocutaneous and visceral abnormalities. Neurological manifestations due to the primary involvement of spinal cord by vascular malformations are rare. We present a young man with HHT associated with a central nervous system arteriovenous malformation and a giant perimedullary fistula, that was manifested as progressive myelopathy. The diagnosis was made coupling magnetic resonance imaging to selective spinal arteriography. The therapeutic option was endovascular treatment by mechanically detachable coils which resulted in full exclusion of the fistula with full improvement of symptoms. During follow-up a stable clinical and morphological outcome was achieved. Clinical manifestations in HHT, with emphasis on neurological symptoms, are reviewed as well as the therapeutic options to deal with giant perimedullary fistula. PMID- 14557904 TI - Cranial MRI in Behcet's disease: 134 examinations of 98 patients. AB - Two observers, blinded to the patient's neurological status, reviewed 134 MRI studies of 98 consecutive patients with Behcet's disease (BD), to define imaging patterns and to look for any relationship between the MRI findings and the timing of the examination in patients with differing courses of disease. There were 43 patients with overt parenchymal central nervous system (CNS) involvement, 22 with attacks and remissions, 15 with secondary progressive and six with primary progressive disease; 14 had raised intracranial pressure (RICP). Of the remaining 41 patients without specific neurological complaints, 16 had abnormalities on examination (silent CNS involvement) and 25 did not. During an acute CNS attack, the most common finding was a large lesion in the brain-stem or basal ganglia, extending to the diencephalon. On MRI performed after remission of an acute attack or during secondary progression, the same sites were affected, but the lesions were smaller or scattered, with less clearly defined margins. In primary progressive disease or silent CNS involvement, the cerebral white matter was most commonly involved, but almost half the MRI studies were normal. The brain parenchyma was abnormal in only one of the patients with RICP. MRI was normal in all but three patients without clinical CNS involvement, in whom it showed a few millimetric white-matter lesions. Brain-stem atrophy was seen in 15 patients examined >1 year after an initial parenchymal CNS episode, with secondary progressive cases predominating. PMID- 14557905 TI - Absolute contraindications in relation to potential drug interactions in outpatient prescriptions: analysis of the first five million prescriptions in 1999. AB - OBJECTIVES: Adverse drug interactions increase morbidity and mortality. To prevent these, situations leading to adverse prescriptions must be clarified. This study quantifies and analyses prescriptions with potential adverse drug interactions in primary health care in the north of France over a 3-month period. METHODS: All prescriptions administered between 1 January and 31 March 1999 were analysed to identify potential interactions amongst drugs appearing on the same prescription sheet. The regional French healthcare database was compiled to further classify contra-indications. RESULTS: There were 5,358,374 prescriptions administered to 44% of the overall population of the Nord-Pas de Calais area (1,754,372 patients/3,990,167 general population). There were 14,390 prescriptions classified as either absolute (26%) or relative contraindications (74%). Nine drug categories accounted for most of the absolute contraindications: dopaminergic antiparkinsonians, neuroleptic agents, migraine treatments (such as ergot alkaloids, sumatriptan and other triptan derivatives), prokinetic drugs (cisapride), antibacterial drugs (macrolides), antifungals (imidazoles), antiarrhythmics, betablockers and analgesics (opioids and floctafenine). In 54% of patients exposed, the incurred risk was either QT prolongation/Torsade de Pointes or antagonism of dopaminergic antiparkinson agents with dopamine receptor antagonists prescribed as antipsychotic agents. CONCLUSIONS: Among a non-selected population of ambulatory outpatients, the number of quarterly prescriptions with contra-indications with potentially harmful drug interaction was 27 in 10,000 prescriptions. This would extrapolate to nearly 200,000 contra-indications on the same-prescription sheets in France in the first quarter of 1999. PMID- 14557906 TI - Effect of inherent epileptic seizures on brain injury after transient cerebral ischemia in Mongolian gerbils. AB - Subthreshold excitotoxic stimuli such as brief cerebral ischemia or chemically induced seizures modulate brain injury resulting from subsequent transient ischemia. Depending on the delay between the two insults, either tolerance or cumulative damage will develop. We were interested whether non-chemically induced inherent epileptic seizures as they occur in Mongolian gerbils have an effect on the outcome of a transient global ischemia, i.e., whether they are an interfering variable in ischemia experiments. Occurrence of spontaneous seizures in adult male gerbils was registered with a video-controlled seizure monitoring system. Bilateral occlusion of common carotid arteries was carried out 2 h or 24 h after the last generalized seizure. After 4 days survival, the extent of ischemia induced neuronal damage and glial activation were assessed in the hippocampus and striatum. No significant difference in the ischemia induced nerve cell loss was observed in cresyl violet stained sections between the 2-h or 24-h interval gerbils. Neuronal expression of endothelial nitric oxide synthase in CA1 disappeared with neuronal degeneration. Distribution and degree of upregulation of glial fibrillary acidic protein as marker for astrocytes did not differ between the two groups. We concluded that non-chemically induced inherent epileptic seizures neither protect the gerbil brain from injury nor augment the degree of damage resulting from transient forebrain ischemia. Thus, inherent epileptic seizures do not influence the outcome of the insult, making the gerbil a reliable model for studies on transient brain ischemia. PMID- 14557907 TI - Early postnatal corticosterone administration regulates neurotrophins and their receptors in septum and hippocampus of the rat. AB - The principal glucocorticoid in rats, corticosterone, interacts with neurons in the limbic system and leads to morphological and behavioral changes. Putative corticosterone-triggered mediators are neurotrophins. In the present study we investigated the effects of early postnatal corticosterone treatment in rats on neurotrophic factors of the nerve growth factor (NGF) family and their receptors. Newborn rats were treated with corticosterone-containing polymers until postnatal day 12. The mRNA and protein levels of the neurotrophins of the NGF family (NGF, BDNF, NT-3 and NT-4/5) and their receptors (trkA, trkB, trkC and p75) were quantified in septum and hippocampus using RT-PCR. In the septal region, we found an unchanged mRNA expression after corticosterone treatment, whereas in the hippocampus there was a general increase in mRNA. Particularly, the gene expression of NGF, NT-3, and the high affinity receptors trkA, trkB and trkC increased significantly. Quantification of the neurotrophin protein levels using an ELISA revealed significant treatment effects for NGF and NT-4/5 in the hippocampus. The present study of corticosterone treatment in young rats demonstrates interactions of steroid hormones with neurotrophic factors and their receptors in the septo-hippocampal system during the first two postnatal weeks. PMID- 14557908 TI - Transient expression of osteopontin mRNA and protein in amoeboid microglia in developing rat brain. AB - To investigate a potential role of osteopontin (OPN) in developing rat brain, the expression of OPN mRNA and protein in the developing rat brain relative to the distribution of brain macrophages was investigated using in situ hybridization and immunohistochemistry, and the phagocytic capability of OPN-expressing cells was accessed using rhodamine isothiocyanate (RhIc) as a tracer. OPN-expressing cells appeared from embryonic day 16. During the first week of postnatal life, OPN-labeled cells increased markedly, and peaked around P7, then declined and had completely disappeared by the end of the second postnatal week. The spatiotemporal distribution pattern of OPN mRNA closely matched that of OPN protein. Their morphology and localization were compared with those of cells expressing the established microglial marker OX-42 in adjacent sections, and double-labeling studies demonstrated that OPN was localized to the amoeboid microglia which stain with the lectin GSI-B4, another marker for microglia. Furthermore, OPN-labeled cells were confirmed to be active phagocytes emitting RhIc fluorescence indicating that the tracer into the brain tissues was engulfed by phagocytosis. Therefore, these results provide the first evidence that OPN is transiently expressed in active brain macrophages in the embryonic and early postnatal brain, and suggest that OPN may contribute to the migration and phagocytic function of brain macrophages in the developing brain. PMID- 14557909 TI - Selective loss of hilar neurons and impairment of initial learning in rats after repeated administration of electroconvulsive shock seizures. AB - Prolonged seizures induced by neurotoxins or intracranial electrical stimulation provoke death of hippocampal neurons, which results in conspicuous learning and memory deficits. We examined whether repeated brief seizures elicited by electroconvulsive shock (ECS) can also deteriorate hippocampal structure and function. Adult Wistar rats were administered six ECS seizures, the first five of which were 24 h apart, whilst the last two were spaced by a 2-h interval. Following a 2-month recovery period, the cognitive status of the animals was assessed using the water maze task. ECS-treated animals were incapable of learning the constant platform position version of this task during the first 4 days of training, but performed similarly to control rats throughout the rest of the acquisition period, on the probe trial, and on the variable platform position and visible platform tasks. The results of the morphological analysis showed that the total number of hippocampal pyramidal neurons and dentate gyrus granule cells were similar in control and ECS-treated rats. However, ECS treatment caused loss of approximately 17% of cells in the hilus of the dentate gyrus, which was accompanied by significant mossy fiber sprouting into the dentate inner molecular layer. In addition, we found that the ECS-induced decrease in the total number of hilar cells was not due to loss of inhibitory interneurons immunoreactive to somatostatin. These findings support the view that ECS-induced seizures can produce a number of morphological and functional changes in the rat hippocampal formation, which qualitatively resemble those previously described in other seizure models. PMID- 14557910 TI - Gap effects on saccade and vergence latency. AB - To explore visual space, we make saccades, vergence, and, most frequently, combined saccade-vergence eye movements. The initiation of saccades is well studied, while that of vergence is less explored. Saccade latency is influenced by the fixation task: when the target appears simultaneously with the offset of the fixation point, latencies tend to be regular, whereas the introduction of a gap period before target onset causes the emergence of express latencies (80- to 120-ms). This study examines in ten normal adults whether the gap paradigm has a similar effect on the latency of vergence and combined eye movements. The second goal is to identify contextual factors that favor the emergence of short latencies, by comparing a condition in which gap and simultaneous trials were performed in separate blocks (pure blocks) with a condition in which the two types of trials were interleaved randomly (mixed blocks). The results are: (1) the gap paradigm reduced similarly (by approximately -30 ms) the mean latency of saccades, convergence, divergence, and both the saccadic and vergence components of combined eye movements; (2) the gap paradigm was responsible for the emergence of 80- to 120-ms latencies for saccades and divergence (pure or combined), but rarely for convergence; (3) inspection of the latency distributions showed that such short latencies formed a clearly distinct population, different from anticipatory responses or regular latencies, for saccades (pure or combined) but not for pure vergence; importantly, distinct express latencies were found also for the convergence and divergence components of combined eye movements; (4) no difference was found for the group of subjects between pure and mixed blocks, but the latter yielded shorter latencies for some subjects, suggesting an idiosyncratic phenomenon. We suggest that distinct express latencies are specific to saccades and could correspond to a specific mode of saccade initiation. Interestingly, the express mode of triggering can be transferred to the vergence component in the ecological condition in which saccade is combined with vergence. PMID- 14557911 TI - Responses of medial olivocochlear neurons. Specifying the central pathways of the medial olivocochlear reflex. AB - Medial olivocochlear (MOC) neurons project to outer hair cells (OHC), forming the efferent arm of a reflex that affects sound processing and offers protection from acoustic overstimulation. The central pathways that trigger the MOC reflex in response to sound are poorly understood. Insight into these pathways can be obtained by examining the responses of single MOC neurons recorded from anesthetized guinea pigs. Response latencies of MOC neurons are as short as 5 ms. This latency is consistent with the idea that type I, but not type II, auditory nerve fibers provide the major inputs to the reflex interneurons in the cochlear nucleus. This short latency also implies that the cochlear-nucleus interneurons have rapidly conducting axons. In the cochlear nucleus, lesions of the posteroventral subdivision (PVCN), but not the anteroventral (AVCN) or dorsal (DCN) subdivisions, produce permanent disruption of the MOC reflex, based on a metric of adaptation of the distortion-product otoacoustic emission (DPOAE). This finding supports earlier anatomical results demonstrating that some PVCN neurons project to MOC neurons. Within the PVCN, there are two general types of units when classified according to poststimulus time histograms: onset units and chopper units. The MOC response is sustained and cannot be produced solely by inputs having an onset pattern. The MOC reflex interneurons are thus likely to be chopper units of PVCN. Also supporting this conclusion, chopper units and MOC neurons both have sharp frequency tuning. Thus, the most likely pathway for the sound-evoked MOC reflex begins with the responses of hair cells, proceeds with type I auditory-nerve fibers, PVCN chopper units, and MOC neurons, and ends with the MOC terminations on OHC. PMID- 14557912 TI - A medullary source of norepinephrine in cat cochlear nuclear complex. AB - Norepinephrine is believed to modulate important functions of the cochlear nuclear complex (CNC) such as the detection of signals in noise and the processing of timing cues. To better understand the impact of the noradrenergic system in the CNC, we used neurotransmitter immunohistochemistry combined with retrograde tract-tracing to identify the noradrenergic cell groups that project to the CNC. Here we present data showing that the CNC receives noradrenergic inputs from the A1 cell group located in the ventrolateral medulla. The projection from A1 to the CNC may be part of a system-wide modulation by the noradrenergic system based on stress and arousal level, or it may be part of a separate circuit that modulates its targets during survival behaviors. PMID- 14557913 TI - Diversity of axonal ramifications belonging to single lateral and medial olivocochlear neurons. AB - A classification of olivocochlear (OC) neurons based on their location in either the lateral or medial region of the superior olivary complex provides a powerful tool for predicting their terminations beneath the inner (IHC) or outer hair cells (OHC) of the cochlea, respectively. Yet the morphology of axonal terminations belonging to single lateral OC (LOC) and medial OC (MOC) neurons, which can provide clues about the functional capabilities of individual efferent neurons, has been relatively unexplored. Following injections of biotinylated dextran amine into regions containing OC neurons in the rat, we reconstructed 19 LOC and 15 MOC axons in surface preparations of the cochlea. Confirming previous studies, LOC axons could be classified as either intrinsic or shell based on the length (short versus long) of their terminal ramifications beneath the IHC. However, intrinsic LOC axons were of two types, those that traveled to the organ of Corti without branching (simple intrinsic) and those that had three or more branches that converged on the same discrete patch of IHCs (converging intrinsic). Regarding shell neurons, we found that they may have as many as four intraganglionic branches that could innervate as much as 41% of cochlear length. Lastly, we found that MOC neurons were extremely diverse, not only in the number of their tunnel-crossing fibers (1-15), but also in both the number of boutons they formed (1-48) and in their basal-apical spans (1-45%). Analysis revealed that the number of tunnel-crossing fibers formed by a given axon was closely related to the total number of its terminal boutons, but not to its cochlear span. Analysis further suggested the existence of two distinct subtypes of MOC neurons on the basis of the number of tunnel-crossing fibers and boutons each possessed: the more common sparsely-branched and the quite rare highly-branched MOC neurons. In conclusion, the variety of axonal ramifications of individual LOC and MOC neurons has functional implications and raises the question of whether the various types of efferent neurons might be subject to selective control by ascending and descending central auditory and possibly non-auditory pathways. PMID- 14557914 TI - Enhanced inhibitory avoidance learning prevents the memory-impairing effects of post-training hippocampal inactivation. AB - Rats were trained on an inhibitory avoidance task to study the effects of post training administration of tetrodotoxin (TTX, which temporarily inactivates neural activity) on memory consolidation. During training, independent groups of rats received either a mild foot shock (0.8 mA) or a stronger (1.0 mA) foot shock. TTX was administered bilaterally into the dorsal hippocampus immediately after training, and memory of the task was measured 48 h later. We corroborated the typical amnesic effect of intrahippocampal infusions of TTX in those rats trained with the mild-intensity foot shock. More importantly, with the stronger foot shock, the same treatment was ineffective in producing amnesia. These results suggest that, after an enhanced learning experience, other brain regions are also activated, which may compensate for the amnesic effect of TTX infusions into the hippocampus. PMID- 14557915 TI - Effects of illusory spatial anisometry in unilateral neglect. AB - Patients with visuospatial neglect tend to underestimate horizontal magnitudes in contralesional space. It has been recently hypothesised that this behaviour might be due to anisometry of space perception, by which horizontal stimuli would be progressively underestimated proceeding from the ipsi towards the contralesional side of space. We investigated the effects of modulating space perception through the Oppel-Kundt illusion (i.e. a filled space is perceived as more expanded than an empty space) on the behaviour of 28 neglect patients and 28 normal subjects. The two groups bisected lines on backgrounds of vertical lines evenly spaced or unevenly spaced, with distances which decreased progressively from one side of the page to the other. On the same backgrounds, they extended segments to the left or to the right so as to double them. Patients also had to cancel targets, the density of which was evenly distributed or horizontally increased from one side of the page to the other. Both groups were prone to the illusion. Neglect bias was modulated by the illusion in the expected direction. It was reduced when the illusion induced a perceptual distortion opposite to that thought to underlie neglect. On bisection and cancellation tasks, illusory effects were greater in patients with higher values of response bias on the Milner Landmark task. These findings, taken together with patients' anatomical data, suggest that a modulation of neglect through a visual illusion can normally be induced in patients with relatively intact visual input processes. PMID- 14557916 TI - Remote activation of referred phantom sensation and cortical reorganization in human upper extremity amputees. AB - Phantom limb sensation, whether painful or not, frequently occurs after peripheral nerve lesions. It can be elicited by stimulating body parts adjacent to the amputation site (referred to as phantom sensation) and it is often similar in quality to the stimulation at the remote site. The present study induced referred phantom sensations in two upper limb amputees. Neuroelectric source imaging (ESI) as well as functional magnetic resonance imaging (fMRI) was used to assess reorganization in primary somatosensory cortex (SI). Whereas recent studies found mislocalization of sensation related to stimulation mainly in regions adjacent and ipsilateral to the amputation site, we report here the elicitation of phantom sensation in the arm by stimulation in the lower body part both ipsi- and contralateral to the amputation in two arm amputees. The fMRI evaluation of one patient showed no shift in the location of the foot whereas ESI revealed major reorganization of the mouth region in primary somatosensory cortex in both patients. These data suggest that cortical structures other than SI might be contributing to the phenomenon of referred sensation. Candidate structures are the thalamus, secondary somatosensory cortex, posterior parietal cortex and prefrontal cortex. PMID- 14557917 TI - Testosterone reinforcement: intravenous and intracerebroventricular self administration in male rats and hamsters. AB - RATIONALE: Anabolic steroids are drugs of abuse. However, the potential for addiction remains unclear. Testosterone induces conditioned place preference in rats and oral self-administration in hamsters. OBJECTIVES: To determine if male rats and hamsters consume testosterone by intravenous (IV) or intracerebroventricular (ICV) self-administration. METHODS: With each nose-poke in the active hole during daily 4-h tests in an operant conditioning chamber, gonad-intact adult rats and hamsters received 50 microg testosterone in an aqueous solution of beta-cyclodextrin via jugular cannula. The inactive nose-poke hole served as a control. Additional hamsters received vehicle infusions. RESULTS: Rats ( n=7) expressed a significant preference for the active nose-poke hole (10.0+/-2.8 responses/4 h) over the inactive hole (4.7+/-1.2 responses/4 h). Similarly, during 16 days of testosterone self-administration IV, hamsters ( n=9) averaged 11.7+/-2.9 responses/4 h and 6.3+/-1.1 responses/4 h in the active and inactive nose-poke holes, respectively. By contrast, vehicle controls ( n=8) failed to develop a preference for the active nose-poke hole (6.5+/-0.5 and 6.4+/ 0.3 responses/4 h). Hamsters ( n=8) also self-administered 1 microg testosterone ICV (active hole:39.8+/-6.0 nose-pokes/4 h; inactive hole: 22.6+/-7.1 nose pokes/4 h). When testosterone was replaced with vehicle, nose-poking in the active hole declined from 31.1+/-7.6 to 11.9+/-3.2 responses/4 h within 6 days. Likewise, reversing active and inactive holes increased nose-poking in the previously inactive hole from 9.1+/-1.9 to 25.6+/-5.4 responses/4 h. However, reducing the testosterone dose from 1 microg to 0.2 microg per 1 microl injection did not change nose-poking. CONCLUSIONS: Compared with other drugs of abuse, testosterone reinforcement is modest. Nonetheless, these data support the hypothesis that testosterone is reinforcing. PMID- 14557918 TI - Drug binding to aromatic residues in the HERG channel pore cavity as possible explanation for acquired Long QT syndrome by antiparkinsonian drug budipine. AB - Budipine is a non-dopaminergic antiparkinsonian drug causing acquired forms of Long QT syndrome (aLQTS). As a consequence, the manufacturer has restricted the use of budipine in patients who exhibit additional risk factors for the development of "Torsades-de-Pointes" tachycardias (TdP). The molecular basis of this serious side effect has not been elucidated yet. Human ether-a-go-go related gene (HERG) channel block being the main cause of drug induced QT prolongation, we investigated the effect of budipine on the rapid component of the delayed rectifier potassium current (I(K(r))) in guinea pig cardiomyocytes and on HERG potassium channels heterologously expressed in Xenopus oocytes. In guinea pig cardiomyocytes, budipine (10 microM) inhibited I(K(r)) by 86% but was without any effect on calcium currents. In Xenopus oocytes, HERG potassium channels were blocked by budipine with an IC(50) of 10.2 microM. Onset of block was fast and block was only slowly and incompletely reversible upon washout. Budipine blocked HERG channels in the open and inactivated state, but not in the closed states. The half-maximal activation voltage was slightly shifted towards more negative potentials. Steady-state inactivation of HERG was also influenced by budipine. Budipine block was neither voltage- nor frequency-dependent. In HERG channel mutants Y652A and F656A, drug affinity was reduced dramatically. Therefore, these two aromatic residues in the channel pore are likely to form a main part of the binding site for budipine. In summary, this is the first study that provides a molecular basis for the budipine-associated aLQTS observed in clinical practice. Furthermore, these findings underline the importance of the aromatic residues Y652 and F656 in the binding of lipophilic drugs to HERG channels. PMID- 14557919 TI - Biomonitoring of aromatic amines VI: determination of hemoglobin adducts after feeding aniline hydrochloride in the diet of rats for 4 weeks. AB - The assessment of the carcinogenic properties of aniline is still controversial. Aniline has, if at all, genotoxic properties but is also acutely toxic and it has been proposed that the hematotoxic effects are responsible for the formation of hemangiosarcomas and fibrosarcomas in the spleen of male rats. As part of a bigger project in which the pathology of male Fischer F344 rats was studied after feeding 10, 30, or 100 mg/kg body weight aniline hydrochloride for 1 and 4 weeks in the diet, the aniline-hemoglobin (Hb) adducts were determined as a biochemical effect marker during those periods. An improved method for the work-up procedure and the adduct analysis was developed for this purpose. The Hb adduct levels increased proportionately with dose after 1 week, which indicates that metabolic activation was not saturated. After 4 weeks of feeding, the adduct levels increased less than proportionately, which suggests that a saturation process is involved. Since it is unlikely that metabolic activation was saturated, the results could be explained by a more rapid clearance of stressed erythrocytes at the carcinogenic dose level. The latter interpretation is supported by other observations which indicate that erythrocytes are damaged dose dependently. A no observed-effect level (NOEL) has not been reached but could be close to the low dose of 10 mg/kg body weight per day. The Hb adduct formation at the low dose, however, indicates that this should not be considered a no-effect level (NEL). The results support the conclusion that hemolytic anemia is an essential prerequisite for aniline toxicity and tumor development, but they do not fully explain the tissue specificity. PMID- 14557920 TI - Meropenem prevents levofloxacin-induced resistance in penicillin-resistant pneumococci and acts synergistically with levofloxacin in experimental meningitis. AB - The aim of the present study was to investigate the potential synergy between meropenem and levofloxacin in vitro and in experimental meningitis and to determine the effect of meropenem on levofloxacin-induced resistance in vitro. Meropenem increased the efficacy of levofloxacin against the penicillin-resistant pneumococcal strain KR4 in time-killing assays in vitro and acted synergistically against a second penicillin-resistant strain WB4. In the checkerboard, only an additive effect (FIC indices: 1.0) was observed for both strains. In cycling experiments in vitro, levofloxacin alone led to a 64-fold increase in the MIC for both strains after 12 cycles. Addition of meropenem in sub-MIC concentrations (0.25 x MIC) completely inhibited the selection of levofloxacin-resistant mutants in WB4 after 12 cycles. In KR4, the addition of meropenem led to just a twofold increase in the MIC for levofloxacin after 12 cycles. Mutations detected in the genes encoding for topoisomerase IV (parC) and gyrase (gyrA) confirmed the levofloxacin-induced resistance in both strains. Addition of meropenem was able to completely suppress levofloxacin-induced mutations in WB4 and led to only one mutation in parE in KR4. In experimental meningitis, meropenem, given in two doses (2 x 125 mg/kg), produced a good bactericidal activity (-0.45 Deltalog10 cfu/ml.h) comparable to one dose (1 x 10 mg/kg) of levofloxacin (-0.44 Deltalog10 cfu/ml.h) against the penicillin-resistant strain WB4. Meropenem combined with levofloxacin acted synergistically (-0.93 Deltalog10 cfu/ml.h), sterilizing the CSF of all rabbits. PMID- 14557921 TI - Pulmonary Mycobacterium xenopi infection in AIDS patients treated with HAART in Hungary. AB - Reported here are three cases of pulmonary Mycobacterium xenopi infection that occurred in AIDS patients in Hungary shortly after starting highly active antiretroviral therapy. In this country, Mycobacterium xenopi is the most common nontuberculous mycobacterial species causing pulmonary mycobacterial infections. Cases of pulmonary Mycobacterium xenopi disease have been described in patients infected with the human immunodeficiency virus infection and in patients with other immunodeficiencies; however, only limited information is currently available concerning the connection between nontuberculous Mycobacterium infection and AIDS in Hungary. This report thus adds useful information regarding the diagnosis, clinical course, and treatment regimens of Mycobacterium xenopi infections in AIDS patients. PMID- 14557922 TI - Rapid identification of yeasts commonly found in positive blood cultures by amplification of the internal transcribed spacer regions 1 and 2. AB - A multiplex PCR method using one universal and eight species-specific primers was developed to rapidly identify eight yeast species found in positive blood cultures. The species-specific primers were designed from the internal transcribed spacer regions 1 and 2 of the rRNA gene, whereas the universal primer was located at the 26S rRNA gene. The eight species were Candida albicans, Candida glabrata, Candida guilliermondii, Candida krusei, Candida lusitaniae, Candida parapsilosis, Candida tropicalis, and Cryptococcus neoformans. The PCR products (116 to 630 bp) were different in length and could be effectively separated and recognized by polyacrylamide gel electrophoresis. By testing 234 positive blood cultures (237 isolates), 234 (98.7%) isolates of the above eight species were correctly identified by the multiplex PCR. The present method is simple to perform and can be completed within 6 h. PMID- 14557923 TI - External quality control of direct antigen tests to detect group A streptococcal antigen. AB - Presented here are the results of an external quality control survey organized by the Swiss Center for Quality Control (CSCQ) to evaluate the performance of direct antigen tests (DATs) widely used in Swiss medical practices and laboratories for the diagnosis of group A streptococcal pharyngitis. Twice yearly over a 4-year period, just over 100 participants were requested to analyze positive, weakly positive and negative samples provided to them by the CSCQ with their routinely used DATs and to send the results to the CSCQ. For 1,620 samples distributed, the CSCQ received 1,484 (91.6%) results obtained with 17 different DATs. The specificity of all DATs for negative samples was >91%. For samples containing abundant group A streptococcal antigen, sensitivities ranged from 59.1% to 95.5%; however, for samples containing low levels of antigen, the sensitivity was much lower for all DATs, ranging from 8.7% to 69.8%. Therefore, negative DAT results should be verified with well-performed cultures in order to assure the optimal care of patients with pharyngitis. PMID- 14557924 TI - Reference values for the SERION classic ELISA for detecting Legionella pneumophila antibodies. PMID- 14557926 TI - Objective evaluation of sensory function in patients with carpal tunnel syndrome using the current perception threshold. AB - The purpose of this study was to determine the sensitivity and specificity for the current perception threshold (CPT) test during clinical examination of idiopathic carpal tunnel syndrome (CTS). We studied 71 patients (102 hands) with CTS; 50 healthy individuals (100 hands) served as a control group. Using a neurometer, we investigated standard CPT values to obtain the sensitivity and specificity of the CPT test in the clinical examination of CTS. In the CTS group, cases that had abnormal values in the range analysis, the ratio analysis, or both were called "true-positive CPTs"; in the control group cases that had normal values in the range and ratio analyses were called "true-negative CPTs." We defined the sensitivity as the percentage of true-positive CPTs in the CTS group and the specificity as the percentage of true-negative CPTs in the control group. The sensitivity of the CPT test using both range and ratio analyses was 73%, and its specificity was 74%. Based on these results we concluded that the CPT test is a reliable quantitative sensory function test. PMID- 14557927 TI - Degenerative osteoarthrosis of tarsometatarsal joints in hallux valgus: a radiographic study. AB - To clarify the pathogenesis of degenerative osteoarthrosis of the tarsometatarsal joints in hallux valgus, we evaluated dorsoplantar and lateral radiographs during weight-bearing in 16 patients (25 feet) with hallux valgus accompanied by degenerative osteoarthrosis of the tarsometatarsal joints and 25 controls (25 feet) with hallux valgus alone. The proximal second metatarsal articular angle (a parameter we devised), the hallux valgus angle, intermetatarsal angle, metatarsal length, sesamoid displacement, calcaneal pitch, and foot length were measured and then evaluation using a mapping system was performed. There were no significant differences in the hallux valgus angle, intermetatarsal angle, sesamoid displacement, calcaneal pitch, or foot length. In the presence of degenerative osteoarthrosis of the tarsometatarsal joints, the second, third, and fourth metatarsals were long, and a large inclination of the proximal articular surface of the second metatarsal and adduction of the first to fourth metatarsals were observed. These findings appeared to be involved in the development of this disorder. PMID- 14557928 TI - Efficacy and limitations of current methods of intraoperative spinal cord monitoring. AB - The purpose of this study was to evaluate the efficacy and reliability of intraoperative spinal monitoring using spinal cord-evoked and compound muscle action potentials. We reviewed 716 cases of spinal monitoring performed over 15 years. The series contained 672 patients with spinal functions that could be monitored intraoperatively; 44 (6.1%) were impossible to record. Based on the 21 impossible-to-record patients, it is evident that missing a serious case such as a Frankel B type spinal cord injury indicates the limitations of the current monitoring methods for stimulating and recording. The monitoring outcomes were true-negative in 652 patients, true-positive in 12, false-negative in four, and false-positive in four. In two of the patients with false-negative results, postoperative myelomere motor paralysis was observed temporarily even though it was possible to record the muscle-evoked potential after electrical stimulation to the brain [Br(E)-MsEP] at the end of the operation. In cases in which the spinal parenchyma or spinal nerve root might be selectively damaged, Br(E)-MsEPs may not diagnose the disorder accurately. By employing multimodal monitoring, it should be possible to eliminate patients with false-negative results and to detect spinal disorders during the early stages, as well as to examine whether the abnormality that had been recorded by a single method is false-positive. PMID- 14557929 TI - Linkage guide for rotational alignment during total knee arthroplasty. AB - A linkage guide was devised for use in conjunction with knee arthroplasty instruments to achieve proper component rotation. The femoral component was rotationally aligned to the surgical epicondylar axis using one guide. The other guides were used after all bone surfaces were cut and soft tissue balancing was completed. A Kirschner wire was guided into the proximal tibial aspect parallel to the sagittal plane of the femoral component with the patella in its normal position and the knee in full extension. The wire was used as a reference to determine tibial component rotation. The relative rotational alignment of 21 knees in 19 patients who had undergone cruciate-retaining total knee arthroplasty utilizing this guide was assessed using the modified Eckhoff method. The results of radiographic analysis were compared with those in a control group of 25 knees where the guide was not employed. The external rotation of the femoral component relative to the tibial component was 1.3 degrees +/- 2.0 degrees (mean +/- SD) in the guided group and 1.1 degrees +/- 4.4 degrees in the control group. The relative rotational angle was significantly more consistent using the guide. Implant malrotation can be reduced using our technique. PMID- 14557930 TI - Results of surgical treatment for lumbar canal stenosis due to degenerative spondylolisthesis: enlargement of the lumbar spinal canal. AB - The purpose of this retrospective study was to analyze clinical results and radiographic findings in patients who underwent surgical enlargement of the lumbar spinal canal combined with resection of the posterosuperior margin underneath the slipping vertebral body for the treatment of lumbar canal stenosis due to degenerative spondylolisthesis. A series of 64 patients who were observed for 3 years or more after operation were examined. The mean age at the time of operation was 64.2 years. The follow-up period was 3-17 years. The Japanese Orthopaedic Association (JOA) score increased from 14.9 points before operation to 25.4 points at the time of the study on average. The general improvement rate was 75.6%. The height and range of motion of the enlarged intervertebral disc were mildly to moderately decreased, and it was found there was a small effect on the adjacent intervertebral disc. On computed tomography, the total level of the enlarged region of the posterosuperior margin increased from 184.4 mm(2) to 339.1 mm(2) on average, but the area of the resected region was 163.3 mm(2) and accounted for 48% of the postoperative area of the spinal canal in the posterosuperior margin. This enlargement of the spinal canal was maintained along the dural canal, and physiological morphology was established. By surgically enlarging the lumbar spinal canal combined with resecting the posterosuperior margin underneath the slipping vertebral body, concomitant repositioning or spinal fixation was unnecessary. PMID- 14557931 TI - Cementless total hip arthroplasty using porous-coated Biomet acetabular cups (Hexloc and Ringloc types). AB - This study examined the mid-term (more than 5 years) results of cementless total hip arthroplasty (THA) using Biomet acetabular cups (locking mechanism: Hexloc type or Ringloc type). A series of 58 patients (70 hips) who had undergone cementless THA at our department were available for inclusion in this study. The average age at surgery was 55.2 years, and the average follow-up was 6 years 10 months. Osteolysis occurred in five cases. In the group with osteolysis, the average annual linear wear was 0.18 mm/year, and in the group without osteolysis it was 0.10 mm/year. The higher linear wear rate in the osteolysis group was statistically significant compared to that in the nonosteolysis group. The age of the patient was closely related to polyethylene (PE) wear in the Hexloc group but not in the Ringloc group. In the Ringloc group there was no close correlation between the thickness of the PE liner and the average annual linear wear, whereas in the Hexloc group the thicker liner had significantly less annual linear wear. It was thought that PE wear of the Hexloc cup was more susceptible to the age of the patient at surgery and the PE thickness than that of the Ringloc cup owing to several structural differences, such as rotational stability and conformity between the metal cup and the PE liner. PMID- 14557932 TI - Minimum 10-year radiographic follow-up of a cementless acetabular component for primary total hip arthroplasty with a bulk autograft. AB - We radiographically investigated 36 hips in 34 patients with osteoarthritis of the hip who had undergone total hip arthroplasty. Their mean age was 59.2 years (range 36-79 years), and the mean follow-up period was 11.2 years (range 10-14 years). The long-term outcome and the chronological changes in the bulk autograft were examined. The acetabular component of the prosthesis was a Lord-type threaded cup with a smooth surface. At follow-up, bone absorption was minor in 17 joints, moderate in 11, and major in 8. The hips with graft coverage of >==20% (group A) had a significantly higher loosening rate than hips with coverage of <<20% (group B) ( P << 0.05). The cup position changed markedly in group A. Our findings indicate that graft coverage should be less than 20% when a bulk graft is used together with a smooth-surfaced cementless cup. PMID- 14557933 TI - Dendritic cell and effector cell infiltration in soft tissue sarcomas with reactive lymphoid hyperplasia. AB - The lymph node is the site of antigen presentation, and dendritic cells are sentinels for anti-tumor immunity. However, little is known about the histological features of lymph nodes and dendritic cells in soft tissue sarcomas. The reactive lymph node and infiltration of dendritic cells or effector cells were studied histologically in 10 soft tissue sarcomas with reactive lymphoid hyperplasia. The cases included four malignant fibrous histiocytomas, two malignant peripheral nerve sheath tumors, one synovial sarcoma, one epithelioid sarcoma, one malignant granular cell tumor, and one liposarcoma. The proportions of the T zone, lymphoid follicle, and lymphoid sinus (which was occupied by cells immunopositive for antibodies against CD3, CD20, or CD68) were 33.4% +/- 11.0%, 6.1% +/- 4.9%, and 13.5% +/- 6.5%, respectively. T zone hyperplasia was observed in all cases, and sinus histiocytosis was found in four. The proportion of the T zone in regional lymph nodes of soft tissue sarcoma patients was significantly higher than that in adult autopsy cases without a cancer history. CD8-, TIA-1-, or granzyme B-positive effector cells were found in each sarcoma tissue. Whereas CD1a-positive dendritic cells were not detected, S-100 protein-positive or CD83 positive dendritic cells were observed in five sarcoma tissues. The coefficient correlation between the numbers of effector cells and dendritic cells positive for CD83 or S-100 protein were demonstrated. Although this is a preliminary report, the present study demonstrated that some soft tissue sarcoma patients showed reactive lymphoid hyperplasia. Furthermore, the association between the infiltration of dendritic cells and that of effector cells was observed in patients with soft tissue sarcomas. PMID- 14557934 TI - Proliferative activity of osteosarcoma cells: comparison of osteoblastic and nonosteoblastic regions. AB - There are two opinions regarding malignancy evaluation by location for osteoblastic osteosarcoma: One is that the nonosteoblastic region is undifferentiated and the degree of malignancy is high; the other is that the osteoblastic region sometimes shows a marked chemotherapy effect, so the degree of biological malignancy is higher. To clarify this point, we compared and examined the difference in growth ability at intratumoral locations within the same tumor using proliferating cell nuclear antigen (PCNA) immunohistochemical staining and argyrophilic nucleolar organizer regions (AgNORs) staining. The subjects were 10 patients with osteoblastic osteosarcoma at the distal region of the femur. Specimens obtained during surgery were stained with PCNA and AgNORs, after which about 2400 tumor cells were counted per field of view, respectively. In all patients, the nonosteoblastic region showed a higher value than the osteoblastic region for both the PCNA labeling index and AgNORs number. A positive correlation was observed between the PCNA labeling index and the AgNORs number. Both the PCNA-labeling index and AgNORs number in the metastatic foci were higher than in the primary lesions. There are differences in proliferative ability in the intratumoral locations within the same osteoblastic osteosarcoma. Moreover, there are differences in proliferative ability between the metastatic foci and the primary lesion. PMID- 14557935 TI - Experimental study on thermal welding for the knee meniscal white zone. AB - It is difficult to repair tears of the meniscus at the white zone by suturing. We considered that there might be an optimal tissue welding temperature when thermally welding the meniscal white zone, so we conducted a thermal welding experiment using bovine and human menisci. The samples prepared after thermal welding were investigated by measuring the tensile strength, analyzing the histological findings of the welded portion and the meniscal parenchyma, and conducting biochemical analyses. In the experiment using human menisci, histological findings were investigated using transmission electron microscopy (TEM). As the findings on the welded tissue in the white zone of the bovine meniscus indicated positive results at 63 degrees C and 69 degrees C, the experiments using human meniscal white zone were conducted at these two temperatures. The highest tensile strength after thermal welding of the human meniscal white zone was 101.4 +/- 2.6 g/cm(2) at 63 degrees C. Although there were no significant differences in the amount of pyridinoline per unit collagen weight [Pyr/Hpr (%)] in the human meniscus after thermal welding between the control group and various temperature groups (55 degrees C, 63 degrees C, 69 degrees C, 75 degrees C), the amount of pyridinoline per unit of collagen weight tended to decline as the welding temperature increased. According to the TEM findings, the internal structure of the nuclei of the cells was preserved in the 63 degrees C group, whereas cells were denatured inside the nuclei in the 69 degrees C group. They exhibited necrosis, making cell regeneration difficult. Therefore, it seems feasible that thermal welding at 63 degrees C can repair tears in the human meniscal white zone. PMID- 14557936 TI - Effect of the rate of prestretching a peripheral nerve on regeneration potential after transection and repair. AB - We tested the influence of nerve stretch injury on nerve regeneration after cutting and suturing. An external fixator was used to lengthen the femur, and consequently the sciatic nerve, progressively, by 3 mm/day (5%/day: group I) or 5 mm/day (9%/day: group II). In both groups the total lengthening was 15 mm, corresponding to approximately 23%-28% elongation of the sciatic nerve. Seven days after initiating nerve lengthening, the external fixator was removed and the nerve was transected and sutured. At 2, 4, 6, or 8 weeks after this suturing, semithin sections were prepared from the sciatic and tibial nerves. Although regeneration of nerve fibers was observed beginning at 2 weeks in all groups, nerve regeneration showed a delay only in group II at 6 weeks, when group I showed nerve regeneration as good as in the unstretched control group. Differences in nerve regeneration after nerve lengthening at different rates were considered the result of mechanical nerve injury, nerve degeneration, ischemia, and fibrosis, all made worse with more rapid lengthening. Nerve lengthening at a more moderate rate (3 mm/day) did not appear to compromise regeneration. PMID- 14557937 TI - Articular cartilage repair of rabbit chondral defect: promoted by creation of periarticular bony defect. AB - This study investigated the stimulatory effect of bone healing on cartilage regeneration. Full-thickness osteochondral defects, 5 mm wide x 3 mm deep, were created in the patellar groove of the right distal end of the femur in 60 mature Japanese white rabbits. The bone defect was made in the medial condyle of the ipsilateral tibia in the experimental group. The control group had only a chondral defect in the patellar groove. At 4, 8, and 12 weeks after surgery, cartilage repair was evaluated macroscopically and histologically using a semiquantitative grading scale. The mean scores of the gross and histological evaluation grade in the experimental group were significantly superior to those in the control group at 8 and 12 weeks ( P << 0.05). These findings suggested that regeneration of articular cartilage defects might be promoted by creation of a periarticular bony defect. PMID- 14557938 TI - Medial collateral ligament and partial anterior cruciate ligament transection: mRNA changes in uninjured ligaments of the sheep knee. AB - Following knee ligament injury, clinical and experimental investigations usually focus on the injured ligament, and uninjured ligaments of the same joint are largely ignored and presumed to remain unchanged. The purpose of this study was to characterize changes in mRNA levels for a relevant subset of molecules in the uninjured knee ligaments following combined unilateral medial collateral ligament (MCL) and partial anterior cruciate ligament (ACL) transection in sheep. Semiquantitative reverse transcription-polymerase chain reaction was performed for collagen types I, III, and V; matrix metalloproteinase-13 (MMP-13); and tissue inhibitor of metalloproteinase-1 for both injured and uninjured knee ligaments at 6 and 12 weeks after injury. Collagen type I, III, and V mRNA levels were significantly increased in MCL scars at 6 weeks as well as in the uninjured lateral collateral ligament and the anteromedial band of the ACL (AM-ACL). MMP-13 mRNA levels were also elevated in the MCL at 6 and 12 weeks and in the AM-ACL 6 weeks after injury. In contrast, significant changes in the posterior cruciate ligament were not detected at either time point, indicating specificity in the transient alterations. These results suggest that following injury, responses occur in uninjured ligaments that are specific although transient in nature. These responses may be an adaptive attempt to preserve function until the scar tissue can stabilize the mechanical environment following injury. PMID- 14557939 TI - Measuring of the compensation of a nerve root in a cervical schwannoma: a case report. AB - A 64-year-old woman experienced numbness and hypesthesia of the right C6 dermatome a year ago. Enhanced magnetic resonance imaging of the cervical spine revealed an enhanced tumor continuing into the foramen from the spinal cord at the C5/6 intervertebral level. It was thought to be an Eden type 2 schwannoma. Right unilateral laminectomy was performed on C5. The tumor was present in the intradural area and arose from the right C6 anterior root. Compound muscle action potentials (CMAPs) from the deltoid, biceps, and extensor carpi radial (ECR) muscles were recorded following electric cervical nerve root stimulation (0.2 ms duration, and 7 mA intensity). CMAPs of large amplitude were obtained from the deltoid, biceps, and ECR muscles following C5 root stimulation, but those following C6 root stimulation were small. As a result it was determined that the right C6 root was not associated with the nerve distribution for these muscles, so it was resected en bloc with the tumor. No apparent loss of motor function was observed. Standard needle electromyography showed no denervation potentials or decrease in motor unit potentials in either the deltoid or biceps muscles. Intraoperative investigation for compensation of nerve root is clinically useful for determining whether resection of a nerve root results in muscle weakness after surgery for a cervical schwannoma. PMID- 14557940 TI - Multiple insufficiency fractures with severe osteoporosis. AB - Multiple insufficiency fracture is a rare injury. We report a 63-year-old woman who spontaneously developed insufficiency fractures at multiple sites including ribs, sacrum, pubis, ischium, acetabulum, metatarsal bone, and femoral neck. The patient had severe osteoporosis with a bone mineral density of 0.267 g/cm(2), although there was no evidence of bone metabolic disease or metastatic bone tumor. Risk factors for osteoporosis in this case were her postmenopausal state and a history of gastrectomy. Interestingly, the serum level of insulin-like growth factor I, recognized as a growth factor that stimulates bone formation, was markedly decreased, and the patient had had viral hepatitis C. It was speculated that the synergistic effects of these disorders might have produced the osteoporosis, leading ultimately to the multiple insufficiency fractures. PMID- 14557941 TI - Acute thoracic myelopathy after a traumatic episode in a patient with neurofibromatosis associated with sharply angular scoliosis: a case report. AB - We treated a 16-year-old boy with acute thoracic myelopathy due to sharply angular scoliosis. The patient underwent posterior decompression and posterior spinal fusion using Luque spinal segmental instrumentation. Postoperatively, spasticity increased, and voluntary movement of the lower extremities disappeared. The patient could walk without assistive devices 5 months postoperatively. As pseudarthrosis at the posterior fusion site was suspected, anterior spinal fusion was performed using a fibular bone strut. His neurologic deficits were completely alleviated, and bony union was obtained. In our case, paraparesis followed relatively mild trauma because the spinal cord was predisposed to injury by continuous mechanical compression, although the cause of the severe temporary deficits following the first operation was not identified. PMID- 14557942 TI - Kienbock's disease with congenital capitohamate coalition in an ulnar ray defect: a case report. AB - We report a patient who developed Kienbock's disease with congenital capitohamate coalition and absence of the little finger: an ulnar ray abnormality of the hand without hypoplasty or defect of the ulna at the forearm. Kienbock's disease combined with an ulnar ray abnormality is a rare situation and the force distribution is unclear. The etiology of the Kienbock's disease is still obscure, so numerous surgical procedures have developed. Capitohamate fusion is one of them, but the case reported here highlights the fact that capitohamate fusion may be a questionable treatment for Kienbock's disease. PMID- 14557943 TI - Computed tomography-guided screw fixation of a sacroiliac joint dislocation fracture: a case report. AB - A 19-year-old woman sustained a vertical shear type pelvic fracture. Sacroiliac fixation using computed tomography (CT)-guided cannulated screws was performed for a left sacroiliac dislocation fracture, and a satisfactory result was obtained over time. Patients who have posterior instability of the lateral compression or vertical shear type do not obtain adequate stability by fixation of the anterior part alone; and they often have persistent residual pain, necessitating internal fixation of the posterior part later. Advantages of CT guided sacroiliac screw fixation include precise evaluation of the degree of reduction and absence of nerve and vascular damage during the time the screw is inserted into the sacral body. This procedure is a useful, safe method owing to its minimal invasiveness in patients with unstable pelvic fractures that are reducible by manual manipulation or traction. PMID- 14557944 TI - Osteoporotic vertebral fracture adjacent to a nonsegmented hemivertebra. AB - A combination of osteoporotic vertebral fractures and congenital spinal deformity is theoretically possible, but there have been no reports on this combination in the literature. We describe a rare case of an osteoporotic vertebral fracture adjacent to the nonsegmented hemivertebra. A 60-year-old postmenopausal woman who did not recall any specific trauma presented with severe back pain. She had markedly decreased bone mineral density and significant lumbar kyphoscoliosis with a nonsegmented hemivertebra between L1 and L2 on radiographs of the lumbar spine. Magnetic resonance imaging (MRI) revealed a vertebral fracture adjacent to the nonsegmented hemivertebra. Laboratory studies showed increased serum bone specific alkaline phosphatase (BAP) and urinary type I collagen crosslinked N telopeptide (NTx). A thoracolumbar brace was applied for 3 months. Daily administration of alendronate normalized her serum BAP and urinary NTx levels. MRI scans of the lumbar spine after 6 months also confirmed normalized signal intensities of the fractured vertebra adjacent to the nonsegmented hemivertebra. The vertebral fracture seemed to be induced by spinal malalignment, increased stress on the adjacent level of the fused segment, and its fragility due to osteoporosis. PMID- 14557945 TI - Genetic analysis of osteoarthritis: toward identification of its susceptibility genes. PMID- 14557946 TI - Controlling mesenchymal stem cell differentiation by TGFBeta family members. AB - Mesenchymal stem cells can differentiate into various tissue types including bone, cartilage, fat, and muscle. Transforming growth factor-Beta (TGFBeta) family members, including TGFBetas and bone morphogenetic proteins (BMPs), play important roles in directing fate decisions for mesenchymal stem cells. TGFBeta can provide competence for early stages of chondroblastic and osteoblastic differentiation, but it inhibits myogenesis, adipogenesis, and late-stage osteoblast differentiation. BMPs also inhibit adipogenesis and myogenesis, but they strongly promote osteoblast differentiation. TGFBeta family members signal via specific serine/threonine kinase receptors and their nuclear effectors, termed Smad proteins. In this review we discuss recent advances in our understanding of the molecular mechanisms by which TGFBeta family members control mesenchymal stem cell differentiation. PMID- 14557947 TI - [Follow-up of patients who seek treatment for grief]. AB - INTRODUCTION: Based on several contributions and studies on evolutive aspects of grief, this study aims to focus on the two-year follow-up of patients from the Community Mental Health Service of Pa rla who came to the clinic in relationship with grief. METHODS: Based upon a previous research, the authors have performed a follow-up study at two years in a sample of 51 patients, carried out by a telephone interview that examined subjective improvement, coping strategies and grieving process, biographical continuity and later life events. RESULTS: Two years after the first visit, most of the patients considered that they had coped well with the loss and were able to resume their normal activities. They ranked their clinical situation as <>. One out of every four reported a partial or marked detention in their biographical course. CONCLUSIONS: In the long term evolution of a visit due to grief, a positive outcome might be expected in the most of the cases. Factors associated with a worst outcome are a problematic bond with the deceased, and the son/daughter loss. Positive outcome was associated with social support and psychotherapy. PMID- 14557948 TI - [Developing a combined predictor measure for early detection of psychosis proneness]. AB - INTRODUCTION: This study falls within research on neurocognitive risk markers of schizophrenia spectrum disorders, and its usefulness for early prevention strategies. METHOD: Two samples of 60 adults and 65 children and adolescents, with some schizophrenia high-risk individuals among them, were studied with the aim of harmonizing different strategies for early detection. One of the most practical aspects of this study was the design of a combined and straightforward measure (different in adults and adolescents) of vulnerability to schizophrenia spectrum disorders (schizotaxia), in contrast to the high availability of neuropsychological measures. For that purpose, several psychometric analyses were made exploring schizotypy prodromes, as well as neurocognitive functions (prefrontal, attentional, working memory and general cognitive functioning tasks) or external markers of these disorders. RESULTS: In keeping with previous research, our results indicate that the more accurate measure of schizotaxia or vulnerability to schizophrenia combines, on the one hand, negative symptoms of schizotypy and, on the other hand, neuropsychological deficits in cognitive frontal functions such as memory, attention and executive functions. CONCLUSIONS: Therefore, the findings suggest that it must be possible to identify normal subjects vulnerable to squizophrenia spectrum disorders, adults o adolescents, because some of them show early neurocognitive deficits and schizotipic symptoms similar to those observed in clinical samples. Actas Esp Psiquiatr 2003;31(5):244 251 PMID- 14557949 TI - [Screening for mild cognitive impairment: usefulness of the 7-Minute Screen Test]. AB - INTRODUCTION: The <<7- Minute Screen>> is a neurocognitive screening test for the detection of Alzheimer's disease (AD) patients in primary care settings. It consists of 4 brief subtests (orientation, memory, visuoconstruction and verbal fluency) and provides a broader neuropsychological profile than other widely used screening tests, The aim of the present study was to study the usefulness of this screening test for the detection of Mild Cognitive Impairment (MCI). METHODS: Thirty-two patients with probable AD (NINCDS-ADRDA criteria), 25 patients with MCI, and 35 healthy control subjects, matched for age and education, underwent a comprehensive neuropsychological battery and the Rio-de-la-Plata version of the 7 Minute Screen. RESULTS. This test showed 93 % sensitivity and 97% specificity in detecting mild-moderate Alzheimer's disease MMSE<24), but it exhibited a substantially decreased sensitivity (28 %) in its ability to detect MCI in AD (MMSE >2 4 ). CONCLUSION: The screening batteries do not replace a m o re compre h e n s i ve neuro psychological assessment. Th ey are useful in detecting patients with mild dementia, but caution must be the rule when considering a diagnosis of MCI. PMID- 14557951 TI - [Opiate dependence users profile. A decade review]. AB - INTRODUCTION: Substance dependence, especially heroin dependence, has become one of the most prevalent psychiatric diseases during recent years. In spite of the numerous related publications, very few articles offer a heroin users profile based on an extensive sample and a broad range of variables of opiate dependent patients in Spain, thus limiting the possibility of generalizing the different findings published. OBJECTIVE: Present a description of an opiate dependence user profile, based on the analysis of a broad range of variables in an extensive population sample. Methodology. An observational, retrospective study, with treatment group (n=1177) and no control group was established. Descriptive and survival techniques were used to analyse different variables (sociodemographic data, medical and psychiatric history, drug consumption record, risk behavior in regards to HIV infection, previous dehabituations, legal situation, treatment carried out for addiction, etc.). RESULTS: We describe a heroine dependent user pattern that coincides in the different variables with the data offered by previous reviews in Spain. CONCLUSIONS: The profile defined can be considered as representative of opiate users in Spain, and it may be used as a reference for future articles. PMID- 14557950 TI - [Olanzapine effects on emotional recognition in treatment refractory schizophrenics]. AB - INTRODUCTION: The main purpose of this study was to determine if olanzapine (OLZ) can improve the ability to recognize emotional expressions, in the facial, prosodic and contextual modalities in treatment refractory schizophrenics (TRS) and, if this could be related to its effects on depressive symptoms. METHOD: 14 TRS participated in the study. The Calgary Depression Scale and tasks for recognition of facial, prosodic and contextual emotions were applied prior to and 8 weeks after consuming OLZ. The results were compared to a control group (CO). RESULTS: TRS obtained lower scores than the CO on the recognition of facial and prosodic emotions. They also showed less empathy to the happiness film and they expressed incongruous answers on the contextual emotions. The TRS increased the number of correct responses for the prosodic recognition of happiness and they showed a reduction in their depressive symptomatology after OLZ treatment. CONCLUSIONS: OLZ caused a decrease of the depressive symptoms and improved the interpretation of positive prosodic affective stimuli, an aspect that may facilitate the social adaptation of TRS. PMID- 14557953 TI - [Pharmacological treatment of substance dependence from a neuroscientific perspective (II): alcohol, benzodiazepines and nicotine]. AB - The second part of this review deals with those neuroscientific aspects specific to the development and maintenance of dependence of three substances, two legal drugs (alcohol and tobacco) and a group of medications with abuse potential, benzodiazepines. Based on this context, the different pharmacological treatments of alcohol dependence, both related to detoxification and dehabituation, are discussed first. Treatment of the benzodiazepine withdrawal syndrome, together with the most outstanding aspects in the recent literature on relapse prevention, are reviewed. The publications on the treatment of nicotine dependence, both on replacement therapies and on bupropion, are analyzed. Finally, a critical reflection of the sources used to conduct this two-part review is done. PMID- 14557952 TI - [The 5-HT1A receptors: from molecular biology to neuropsychiatric symptoms]. AB - Among the multiple serotonin receptors identified to date, the 5 hydroxytryptamine (5-HT)1A subtype is among the best known because selective ligands have been available for more than 15 years. Radioactive derivatives of these ligands make it possible to demonstrate the presence of 5-HT1A binding sites mainly in the limbic areas and in the raphe nuclei in the brain, where they correspond to post-synaptic receptors and pre-synaptic autoreceptors respectively. On stimulation of 5-HT1A autoreceptors, they regulate serotonin release in the distal regions of the neuron by inhibitory firing activity. In this way, they help to maintain the serotonin in the terminal regions at physiological levels, which favors correct neuronal functioning. This review article summarizes key data on localization, study technique, molecular biology, signal transduction, differential functional properties of pre-synaptic versus post-synaptic 5-HT1A receptors, and behavioral effects and clinical correlates of their activation, especially cognitive and emotional responses. Mention is made about the role of these receptors in the neurogenesis process of certain brain areas and of the possible clinical and therapeutic implications of these processes. PMID- 14557954 TI - [Cannabis dependence: clinical implications. Based and one case]. AB - Cannabis has been commonly regarded as an innocuous drug, and has been used for centuries due to its multiple therapeutic effects (analgesic, anticonvulsant, sedative, antidepressant, etc.). However, growing evidence in recent years highlights the risk of adverse effects secondary to dependence and withdrawal syndrome. We report a case of severe cannabis dependence, that led to a variety of psychopathological, cognitive and somatic symptoms. This is compared with the bibliography and we analyze the influence of a possible underlying psychiatric disease. The adverse effects include affective and eating disorders, and the possibility of seizure induction. A novel therapeutic strategy in cannabis dependence is reported. Key words: Cannabis. Mania. Bulimic behaviors. PMID- 14557955 TI - [Familiar Munchausen syndrome: an unusual presentation]. AB - Munchausen syndrome is a subtype included in factitious disorders in which somatic signs and symptoms predominate. The patient self-inflicts damage with the only objective of assuming a <1 year after the onset of CSD. Onset of CSD in patients with an IgG titer with an optical density of > or =1.0 occurred within the prior 12 months. No association was found between antibody titers or their kinetics and the clinical manifestations or duration of disease. EIA allows for the identification of atypical manifestations of CSD that were unrecognized before the use of serological assays. Complete recovery from these manifestations may take months. Results of this study provide additional data supporting the utility of EIA in the serodiagnosis of CSD. PMID- 14557958 TI - Protective efficacy of a second pneumococcal conjugate vaccine against pneumococcal acute otitis media in infants and children: randomized, controlled trial of a 7-valent pneumococcal polysaccharide-meningococcal outer membrane protein complex conjugate vaccine in 1666 children. AB - To assess the efficacy of a 7-valent pneumococcal polysaccharide-meningococcal outer membrane protein complex conjugate vaccine (PncOMPC) against acute otitis media (AOM), 1666 infants were randomly assigned to receive either PncOMPC or control vaccine (hepatitis B vaccine) at 2, 4, 6, and 12 months of age. Of the 835 children assigned to receive PncOMPC, 187 received a 23-valent pneumococcal polysaccharide vaccine (PncPS) at 12 months of age instead. Whenever AOM was diagnosed, middle ear fluid was aspirated for bacterial culture. In the PncOMPC and control groups, there were 110 and 250 AOM episodes, respectively, in children between 6.5 and 24 months of age that could be attributed to vaccine serotypes, which indicates a vaccine efficacy of 56% (95% confidence interval, 44%-66%). The serotype-specific efficacy ranged from 37% for 19F to 82% for 9V. The 2 boosters seemed to provide equal protection against AOM, but PncPS induced markedly higher antibody concentrations. The efficacy of PncOMPC was comparable to that of the recently licensed pneumococcal conjugate vaccine. PMID- 14557959 TI - Azithromycin found to be comparable to levofloxacin for the treatment of US travelers with acute diarrhea acquired in Mexico. AB - Increased drug resistance among enteropathogens is an emergent problem in travelers' diarrhea. This randomized, double-blind trial was conducted in Guadalajara, Mexico, during the summers of 1999-2001 to compare azithromycin with levofloxacin for the treatment of travelers' diarrhea. A total of 217 US adults were randomized to receive a single oral dose of azithromycin (1000 mg; 108 persons) or levofloxacin (500 mg; 109 persons), with a follow-up period of 4 days. Three patients in each group dropped out of the study. The median time between initiation of therapy and passage of the last unformed stool (azithromycin group, 22.3 h; levofloxacin group, 21.5 h) and the number of unformed stools passed during the 4-day follow-up period (azithromycin group, 6.5; levofloxacin group, 5.5) were similar. Treatment failure occurred in 10 patients (9.5%) receiving azithromycin and 8 patients (7.5%) receiving levofloxacin. Possible minor, self-limiting adverse events occurred in 57 patients in each treatment group. Azithromycin was found to be a safe and effective alternative to levofloxacin for the treatment of acute travelers' diarrhea in US adult travelers to Mexico. PMID- 14557960 TI - Attributable mortality of nosocomial candidemia, revisited. AB - We reexamined the attributable mortality of nosocomial candidemia 15 years after a retrospective cohort study performed at our hospital demonstrated an attributable mortality of 38%. For all episodes of nosocomial candidemia between 1 July 1997 and 30 June 2001, we matched control patients with case patients by age, sex, date of hospital admission, underlying disease(s), length of time at risk, and surgical procedure(s). We analyzed 108 matched pairs. There were no statistically significant differences in age, sex, underlying disease(s), time at risk, surgical procedure, or vital signs at admission between cases and controls. The crude mortality among case patients was 61% (66 of 108 patients), compared with 12% (13 of 108) among control patients, for an attributable mortality of 49% (95% CI, 38%-60%). Nosocomial candidemia is still associated with an extremely high crude and attributable mortality--much higher than that expected from underlying disease alone. PMID- 14557961 TI - Thrice-weekly clarithromycin-containing regimen for treatment of Mycobacterium kansasii lung disease: results of a preliminary study. AB - We initiated a prospective trial of an intermittent clarithromycin-containing regimen for the treatment of patients with Mycobacterium kansasii lung disease. Eighteen patients (10 men and 8 women) with M. kansasii lung disease received a regimen consisting of 500-1000 mg of clarithromycin, 25 mg/kg ethambutol, and 600 mg of rifampin 3 times per week. The primary treatment end point was a 12-month period during which sputum cultures were sterile while the patient was receiving therapy. Four male patients were lost to follow-up, but all of the remaining patients successfully completed therapy without significant drug-related adverse events. The mean time (+/- standard deviation [SD]) to sputum conversion was 1.0+/-0.9 months, and the mean duration (+/-SD) of therapy was 13.4+/-0.9 months. No patient who successfully completed therapy had relapsed after a mean (+/-SD) of 46+/-8.0 months. Clarithromycin- and rifampin-containing regimens offer the possibility of effective short-course and intermittent treatment of M. kansasii lung disease. PMID- 14557962 TI - Neisseria meningitidis carriage during an outbreak of serogroup C disease. AB - During 2001, an outbreak of serogroup C meningococcal disease led to immunization of individuals aged 13-29 years in Abbotsford, British Columbia, Canada. This study addresses the distribution of Neisseria meningitidis carriage in this population and the implications of that distribution for the targeting of the immunization campaign. Pharyngeal swabs were obtained at immunization from 2004 people. Colonies were identified and serogrouped using standard agglutination methods and by PCR. Isolates were characterized using pulsed-field gel electrophoresis (PFGE). The prevalence of N. meningitidis carriage was 153 carriers per 2004 subjects (7.6%; 95% confidence interval, 6.5%-8.9%). Only 6 (4%) of the isolates from these carriers were found to be serogroup C by agglutination or PCR testing, and all of these were from individuals within the age group targeted for immunization. Only 1 of these 6 isolates was found to be identical to the outbreak strain by PFGE. The observation that a virulent strain is not circulating widely suggests the possibility of low background immunity in the population at risk and emphasizes the importance of vaccination in controlling epidemic group C meningococcal disease. PMID- 14557963 TI - Group A streptococcal infections in Sweden: a comparative study of invasive and noninvasive infections and analysis of dominant T28 emm28 isolates. AB - Surveillance of group A streptococcus (GAS) infections in Sweden during 1996-1997 indicated that T28 isolates were dominant, whereas T1M1 infections were uncommon. Circulating T28 isolates were nearly all emm28, MLST52, and these clones had also been prevalent 10 years earlier. Isolates from invasive and noninvasive infections were of similar types and prevalences. The average national incidence of invasive episodes was 2.9/100,000 population but varied between 0 and 8.3/100,000 population in different counties. It increased markedly with age, reaching 22.9 episodes/100,000 among people aged > or =90 years. The incidence of puerperal sepsis was higher than expected (22.4/100,000 of those at risk), with 1 death. Overall mortality was 16% and was associated with preexisting chronic disease (P=.002). Streptococcal toxic shock syndrome (STSS) developed in approximately 15% of patients with invasive episodes, with a mortality rate of 45%. The use of nonsteroidal anti-inflammatory drugs was not found to be associated with the development of STSS. PMID- 14557964 TI - Relapse of type A beta-lactamase-producing Staphylococcus aureus native valve endocarditis during cefazolin therapy: revisiting the issue. AB - Our experience with a patient with methicillin-susceptible Staphylococcus aureus aortic native valve endocarditis, who had a relapse involving fever and positive blood culture results while receiving cefazolin, led us to evaluate this organism's ability to hydrolyze cefazolin at high inocula, a previously well documented phenomenon. Analysis of the infecting strain disclosed a high minimum inhibitory concentration of cefazolin when a large inoculum was used, as well as rapid and complete cefazolin degradation, which was associated with regrowth in a time-kill experiment. DNA sequencing of the beta-lactamase gene showed that it was identical to that of the S. aureus type A beta-lactamase, known to efficiently inactivate cefazolin. A word of caution is given regarding the use of this antibiotic for treatment of endocarditis caused by this type of S. aureus isolate. PMID- 14557965 TI - Hypermutation as a factor contributing to the acquisition of antimicrobial resistance. AB - Contrary to what was thought previously, bacteria seem to be, not merely spectators to their own evolution, but, through a variety of mechanisms, able to increase the rate at which mutations occur and, consequently, to increase their chances of becoming resistant to antibiotics. Laboratory studies and mathematical models suggest that, under stressful conditions, such as antibiotic challenge, selective pressure favors mutator strains of bacteria over nonmutator strains. These hypermutable strains have been found in natural bacterial populations at higher frequencies than expected. The presence of mutator strains in the clinical setting may indicate an enhanced risk of acquiring antibiotic resistance through mutational and recombinational events. In addition, some antibiotics are inducers of mechanisms that transiently increase the mutation rate, and thus probably act, not only as mere selectors of antibiotic resistant clones, but also as resistance promoters. PMID- 14557966 TI - Gatifloxacin, gemifloxacin, and moxifloxacin: the role of 3 newer fluoroquinolones. AB - Gatifloxacin, gemifloxacin, and moxifloxacin are the newest fluoroquinolones and show excellent in vitro activity against a wide variety of respiratory tract pathogens, many gram-negative aerobic organisms, and Bacteroides fragilis. These agents may be administered as oral and/or intravenous formulations with excellent bioavailability. The pharmacodynamics of these 3 new fluoroquinolones is more favorable than that of levofloxacin or ciprofloxacin for Streptococcus pneumoniae. All 3 agents are approved for the treatment of acute exacerbation of chronic bronchitis and community-acquired pneumonia. In addition, gatifloxacin and moxifloxacin are approved for the treatment of sinusitis. The toxicity of these 3 agents appears to be similar to that of the other fluoroquinolones in terms of gastrointestinal and central nervous system disturbances. All 3 agents have a low risk of phototoxicity, but gemifloxacin is associated with an increased risk of skin rash that is not a photoreaction. These agents can be useful for treatment of bacterial respiratory tract infections in patients who are allergic to beta-lactams, but caution must be exercised to avoid the potential for selection of widespread resistance, which may occur with indiscriminate use. PMID- 14557967 TI - Viral load and CD4+ T lymphocyte response to highly active antiretroviral therapy in human immunodeficiency virus type 1-infected children: an observational study. AB - An observational study was performed involving 95 children with vertically transmitted human immunodeficiency virus type 1 infection to assess the sustainability of undetectable viral loads (VLs) and increased CD4+ T lymphocyte percentages after 48 months of highly active antiretroviral therapy (HAART). The median time to achieve a 10% increase in the CD4+ T lymphocyte percentage was 11.01 months. The median time to achieve an undetectable VL was 6.4 months. At the end of the study, 64.2% of the children had achieved an undetectable VL. Of the patients with an initial VL of >3.6 log10 copies/mL, 74.7% had a decrease in the VL of 1 log10 copies/mL. By contrast, of the patients who presented with an initial VL of >4.6 log10 copies/mL, 37.9% had a decrease of >2 log10 copies/mL. Higher VL at baseline, antiretroviral therapy regimens received before HAART, and multiple drug switches while receiving antiretroviral therapy were all inversely associated with an undetectable VL. A CD4+ T lymphocyte percentage of >25% was directly associated with undetectable VL during the follow-up period. In conclusion, first-line HAART induces beneficial virological and immunological outcome responses in children. PMID- 14557968 TI - Danish bacille Calmette-Guerin vaccine-induced disease in human immunodeficiency virus-infected children. AB - An analysis of isolates of Mycobacterium tuberculosis complex was performed to determine the prevalence of bacille Calmette-Guerin (BCG) disease among human immunodeficiency virus (HIV)-infected children. Speciation was done with polymerase chain reaction; 183 isolates from mycobacterial cultures for 49 HIV infected patients were analyzed. The Danish Mycobacterium bovis BCG strain was isolated from 5 patients. No cases of Tokyo M. bovis BCG strain disease were detected. All patients were asymptomatic at birth, <12 months of age, and severely immunodeficient at presentation. Four patients had regional axillary adenitis ipsilateral to the vaccination site, and 2 had pulmonary BCG disease. Two patients with regional BCG disease had simultaneous pulmonary M. tuberculosis infection. Although chest radiographic features were similar to those seen in patients with tuberculosis, BCG disease should be considered in HIV-infected infants with right axillary adenitis ipsilateral to the vaccination site. Young, symptomatic, HIV-infected infants are at risk for BCG-related complications. Controlled, population-based studies are needed to assess the risk of BCG in HIV infected children. PMID- 14557969 TI - A prospective, randomized trial examining the efficacy and safety of clarithromycin in combination with ethambutol, rifabutin, or both for the treatment of disseminated Mycobacterium avium complex disease in persons with acquired immunodeficiency syndrome. AB - This multicenter, randomized, open-label phase 3 clinical trial compared the safety and efficacy of 3 clarithromycin-containing combination regimens for the treatment of disseminated Mycobacterium avium complex (MAC) disease in persons with acquired immunodeficiency syndrome. A total of 160 eligible patients with bacteremic MAC disease were randomized to receive clarithromycin with either ethambutol (C+E), rifabutin (C+R), or both (C+E+R) for 48 weeks. After 12 weeks of treatment, the proportion of subjects with a complete microbiologic response was not statistically significantly different among treatment arms: the proportion was 40% in the C+E group, 42% in the C+R group, and 51% in the C+E+R group (P=.454). The proportion of patients with complete or partial responses who experienced a relapse while receiving C+R (24%) was significantly higher than that of patients receiving C+E+R (6%; P=.027) and marginally higher than that of patients receiving C+E (7%; P=.057). Subjects in the C+E+R group had improved survival, compared with the C+E group (hazard ratio [HR], 0.44; 95% confidence interval [CI], 0.23-0.83) and the C+R group (HR, 0.49; 95% CI, 0.26-0.92). PMID- 14557970 TI - Epstein-Barr virus DNA loads in adult human immunodeficiency virus type 1 infected patients receiving highly active antiretroviral therapy. AB - Patients with human immunodeficiency virus type 1 (HIV-1) infection are at high risk of developing Epstein-Barr virus (EBV)-associated lymphoma. However, little is known of the EBV DNA loads in patients receiving highly active antiretroviral therapy (HAART). Using a real-time quantitative polymerase chain reaction assay, we demonstrated that significantly more HIV-1-infected patients receiving HAART than HIV-1-uninfected volunteers had detectable EBV DNA in blood (57 [81%] of 70 vs. 11 [16%] of 68 patients; P=.001) and saliva (55 [79%] of 68 vs. 37 [54%] of 68 patients; P=.002). The mean EBV loads in blood and saliva samples were also higher in HIV-1-infected patients than in HIV-1-uninfected volunteers (P=.001). The frequency of EBV detection in blood was associated with lower CD4+ cell counts (P=.03) among HIV-1-infected individuals, although no differences were observed in the EBV DNA loads in blood or saliva samples in the HIV-1-infected group. Additional studies are needed to determine whether EBV-specific CD4+ and CD8+ cells play a role in the pathogenesis of EBV in HIV-1-infected patients receiving HAART. PMID- 14557971 TI - Human immunodeficiency virus type 1-related pulmonary Mycobacterium xenopi infection: a need to treat? AB - We report treatment decisions and outcomes for 20 patients who were infected with human immunodeficiency virus type 1 (HIV-1) and were receiving highly active antiretroviral therapy (HAART) who had respiratory symptoms and from whom Mycobacterium xenopi was isolated. All patients also had coexisting pulmonary pathologic conditions. The median blood T cell CD4 count was 37 cells/microL (range, 2-480 cells/microL). Fifteen of 20 patients received no antimycobacterial therapy and remain healthy after a median of approximately 4 years of follow-up, and 2 patients required treatment specifically for M. xenopi infection, both showing clinical improvement. We conclude that pulmonary M. xenopi isolation in HIV-1 patients receiving HAART does not usually require specific treatment. PMID- 14557972 TI - Failure of clindamycin treatment of methicillin-resistant Staphylococcus aureus expressing inducible clindamycin resistance in vitro. AB - We report a case of a surgical site infection caused by clindamycin-susceptible, erythromycin-resistant methicillin-resistant Staphylococcus aureus (MRSA) that did not respond to treatment with clindamycin. The MRSA isolate obtained after treatment was resistant to clindamycin but was found to be identical by pulsed field gel electrophoresis to the clindamycin-susceptible isolate obtained before treatment. A post hoc erythromycin-induction test (D test) confirmed the presence of in vitro inducible macrolide-lincosamide-streptogramin B resistance (iMLS) in the pretreatment isolate. Erythromycin induction testing confirmed in vitro iMLS in 90 (56%) of 161 erythromycin-resistant, clindamycin-susceptible clinical S. aureus isolates overall and in a significantly higher proportion (78%) of methicillin-susceptible S. aureus isolates from pediatric patients. Our clinical laboratory currently tests all S. aureus isolates for iMLS before reporting clindamycin susceptibility. PMID- 14557973 TI - Serum concentrations of pneumococcal anticapsular antibodies in children with pneumonia associated with Streptococcus pneumonia infection. AB - The levels of specific IgG antibody to pneumococcal capsular polysaccharides were investigated in 182 children, aged 2-5 years, who were hospitalized for community acquired pneumonia, including 55 (30.2%) with evidence of acute pneumococcal infection. Results show that children with concentrations of specific IgG antibody that would protect against invasive disease do not seem to be protected against pneumonia associated with pneumococcal infection. PMID- 14557974 TI - Donor-related coccidioidomycosis in organ transplant recipients. AB - Most cases of coccidioidomycosis in organ transplant recipients arise from either primary infection with Coccidioides immitis after environmental exposure or from reactivation of latent infection. Herein, we report 2 cases of rapidly fatal, disseminated coccidioidomycosis that occurred in organ transplant recipients who had never lived in or visited an area where C. immitis is endemic. Both subjects had received a transplanted organ from the same donor, an individual with unrecognized active coccidioidomycosis at the time of his death. PMID- 14557975 TI - High incidence of pertussis among Hajj pilgrims. AB - Prolonged cough occurs in a large proportion of the 2 million pilgrims who participate in the annual Hajj in Saudi Arabia. In a prospective seroepidemiological study to determine the incidence of pertussis among 358 adult pilgrims, 5 (1.4%) were found to have acquired pertussis (defined as prolonged cough and a >4-fold increase in the level of immunoglobulin G to whole-cell pertussis antigen). Of the 40 pilgrims who had no pre-Hajj immunity to pertussis, 3 (7.5%) acquired pertussis. Administration of acellular pertussis vaccine to pilgrims before the Hajj should be considered to address this problem. PMID- 14557976 TI - Amprenavir resistance imparted by the I50V mutation in HIV-1 protease can be suppressed by the N88S mutation. PMID- 14557977 TI - Linezolid and serotonin toxicity. PMID- 14557978 TI - Isolated antibody to hepatitis B core antigen in individuals infected with HIV-1. PMID- 14557980 TI - Coinfection in patients with lyme disease: how big a risk? PMID- 14557982 TI - Successful booster antibody response up to 54 months after single primary vaccination with virosome-formulated, aluminum-free hepatitis A vaccine. AB - This study demonstrates that a booster dose of the virosome-formulated, aluminum free hepatitis A vaccine Epaxal (Berna Biotech) is highly immunogenic in subjects who received a single primary dose of this vaccine 18-54 months earlier. There were no significant differences in geometric mean antibody titers (GMTs) among subjects who received the booster dose 18-29 months (GMT, 2330 mIU/mL), 30-41 months (GMT, 2395 mIU/mL), or 42-54 months (GMT, 2432 mIU/mL) after primary vaccination, indicating that delays in the administration of booster vaccination do not lead to a loss of immunogenicity. PMID- 14557984 TI - Case report and literature review of late-onset group B streptococcal disease manifesting as necrotizing fasciitis in preterm infants: is this a new syndrome? AB - We describe a 5-month-old preterm female infant who presented with necrotizing fasciitis involving the face and neck caused by group B streptococcus (GBS). Because of the extent and anatomic location of the necrosis, surgical debridement was delayed for 16 days, but the infant survived. Review of the literature demonstrated that 3 of the 10 previously reported cases of necrotizing fasciitis caused by GBS involved preterm infants and that 2 of these cases also involved the head and neck. PMID- 14557983 TI - A fatal case of West Nile virus infection in a bone marrow transplant recipient. AB - West Nile virus (WNV) can cause severe, potentially fatal neurological illnesses, which include encephalitis, meningitis, Guillain-Barre syndrome, and anterior myelitis. Because of the short viremic phase, WNV infection is most commonly diagnosed by detection of immunoglobulin M antibody to WNV in serum or cerebrospinal fluid (CSF). We describe a patient with T cell lymphoma who had undergone a T cell-depleted bone marrow transplantation and developed fatal WNV infection. The results of serological tests of blood samples and of CSF tests were negative. Diagnosis was made postmortem by a positive result of reverse transcriptase polymerase chain reaction (ABI 7700; TaqMan) for WNV in stored CSF and serum samples. PMID- 14557985 TI - A major susceptibility locus on chromosome 22q12 plays a critical role in the control of kala-azar. AB - Kala-azar (KA) is a life-threatening protozoal disease caused by Leishmania parasites (L. donovani, L. chagasi, and L. infantum). The disease, which is also called "visceral leishmaniasis," is prevalent in Africa, South America, Asia, and the Mediterranean basin. Epidemics occur periodically, killing a large number of infected individuals. Factors determining whether a patient remains asymptomatic or develops KA are still largely unknown. In a previous study that was performed during an outbreak of KA in a village on the Ethiopian-Sudanese border, we showed that KA was more frequent in certain families and ethnic groups, thereby suggesting that host genetic factors play an important role in the development of the disease. Here, we report the results of a genomewide linkage study performed on 63 Sudanese families selected from the most affected ethnic group and including 169 children with KA. Significant linkage (LOD score 3.50 [P=3x10-5] in all patients; LOD score 3.90 [P=10-5] in patients who were affected early in the outbreak) was obtained with markers on chromosome 22q12. These results are the first evidence of a major genetic effect on the development of human KA. They may lead to identification of genes critical in the pathogenesis of this disease and to new therapeutic interventions against this parasite, which is developing resistance to available drugs. PMID- 14557986 TI - Logistic regression of family data from retrospective study designs. AB - We wish to study the effects of genetic and environmental factors on disease risk, using data from families ascertained because they contain multiple cases of the disease. To do so, we must account for the way participants were ascertained, and for within-family correlations in both disease occurrences and covariates. We model the joint probability distribution of the covariates of ascertained family members, given family disease occurrence and pedigree structure. We describe two such covariate models: the random effects model and the marginal model. Both models assume a logistic form for the distribution of one person's covariates that involves a vector beta of regression parameters. The components of beta in the two models have different interpretations, and they differ in magnitude when the covariates are correlated within families. We describe ascertainment assumptions needed to estimate consistently the parameters beta(RE) in the random effects model and the parameters beta(M) in the marginal model. Under the ascertainment assumptions for the random effects model, we show that conditional logistic regression (CLR) of matched family data gives a consistent estimate beta(RE) for beta(RE) and a consistent estimate for the covariance matrix of beta(RE). Under the ascertainment assumptions for the marginal model, we show that unconditional logistic regression (ULR) gives a consistent estimate for beta(M), and we give a consistent estimator for its covariance matrix. The random effects/CLR approach is simple to use and to interpret, but it can use data only from families containing both affected and unaffected members. The marginal/ULR approach uses data from all individuals, but its variance estimates require special computations. A C program to compute these variance estimates is available at http://www.stanford.edu/dept/HRP/epidemiology. We illustrate these pros and cons by application to data on the effects of parity on ovarian cancer risk in mother/daughter pairs, and use simulations to study the performance of the estimates. PMID- 14557987 TI - Polygenic inheritance of breast cancer: Implications for design of association studies. AB - Susceptibility to breast cancer is likely to be the result of susceptibility alleles in many different genes. In particular, one segregation analysis of breast cancer suggested that disease susceptibility in noncarriers of BRCA1/2 mutations may be explicable in terms of a polygenic model, with large numbers of susceptibility polymorphisms acting multiplicatively on risk. We considered the implications for such a model on the design of association studies to detect susceptibility polymorphisms, in particular the efficacy of utilizing cases with a family history of the disease, together with unrelated controls. Relative to a standard case-control association study with cases unselected for family history, the sample size required to detect a common disease susceptibility allele was typically reduced by more than twofold if cases with an affected first-degree relative were selected, and by more than fourfold if cases with two affected first-degree relatives were utilized. The relative efficiency obtained by using familial cases was greater for rarer alleles. Analysis of extended families indicated that the power was most dependent on the immediate (first-degree) family history. Bilateral cases may offer a similar gain in power to cases with two affected first-degree relatives. In contrast to the strong effect of family history, varying the ages at diagnosis of the cases across the range of 35-65 years did not strongly affect the power to detect association. These results indicate that association studies based on cases with a strong family history, identified for example through cancer genetics clinics, may be substantially more efficient than population-based studies. PMID- 14557988 TI - Genotype-based association test for general pedigrees: the genotype-PDT. AB - Many family-based tests of linkage disequilibrium (LD) are based on counts of alleles rather than genotypes. However, allele-based tests may not detect interactions among alleles at a single locus that are apparent when examining associations with genotypes. Family-based tests of LD based on genotypes have been developed, but they are typically valid as tests of association only in families with a single affected individual. To take advantage of families with multiple affected individuals, we propose the genotype-pedigree disequilibrium test (geno-PDT) to test for LD between marker locus genotypes and disease. Unlike previous tests for genotypic association, the geno-PDT is valid in general pedigrees. Simulations to compare the power of the allele-based PDT and geno-PDT reveal that under an additive model, the allele-based PDT is more powerful, but that the geno-PDT can have greater power when the genetic model is recessive or dominant. Perhaps the most important property of the geno-PDT is the ability to test for association with particular genotypes, which can reveal underlying patterns of association at the genotypic level. These genotype-specific tests can be used to suggest possible underlying genetic models that are consistent with the pattern of genotypic association. This is illustrated through an application to a candidate gene analysis of the MLLT3 gene in families with Alzheimer disease. The geno-PDT approach for testing genotypes in general family data provides a useful tool for identifying genes in complex disease, and partitioning individual genotype contributions will help to dissect the influence of genotype on risk. PMID- 14557989 TI - Detecting low-quality markers using map expanders. AB - Genetic marker data play a crucial role in gene mapping, and genotyping errors may have substantial influence on the power to detect and the precision to locate disease loci. Statistical methods can identify individuals, markers, or pedigrees with a high likelihood of containing genotyping errors, and the putative erroneous genotypes can then be rechecked and either verified, removed, or corrected to reduce the loss of power introduced by errors. We present a method to identify genetic markers with a high genotyping error rate. Genotyping errors are likely to appear as double recombinations which expand the genetic map around the marker. Markers flagged as map expanders (i.e., having an excessive number of double recombinations) can then be reread or regenotyped, or a replacement marker of higher quality can be used instead. The proposed method can be applied to any type of pedigree. Simulation studies of nuclear pedigrees and sib-pairs show that the proposed method generally has high power to identify map expanders when the set of markers is reasonably dense (average intermarker distance of 5 cM), even when the nominal genotyping error rate is low (2%). Not surprisingly, the power to detect map expanders increases with marker heterozygosity and genotyping error rate, and is reduced with increasing intermarker distance. When the method was applied to a real dataset consisting of 56 nuclear pedigrees genotyped for 20 microsatellite markers on chromosome 4, the method diagnosed three markers as map expanders. Subsequent examination of these markers proved that they all had high genotyping error frequencies. PMID- 14557990 TI - Analysis of twin data ascertained through probands: the double-entry approach. AB - Twin pairs are sometimes included in studies because at least one of them is a proband, and conventionally the analysis of the data is based on the conditional distribution of the co twin given the proband. In the case of more than one proband in each pair, an often used "ad hoc" method of analysis is to allow each twin to act as proband as well as co twin. An example of this is the pro band wise concordance, which is used as an estimate of the case wise concordance under incomplete ascertainment. In this paper, we show that the method of double entry under a regularity condition provides consistent estimates of parameters, but that conventional standard errors as well as a correction suggested by Stevenson et al. will be wrong. Instead, we recommend the use of an easily calculated sandwich estimator. We illustrate the method with twin data from a study of the genetic component in hand eczema. PMID- 14557992 TI - "Father" of experimental microsurgery: Dr. Sun Lee. PMID- 14557993 TI - Transplantation and microsurgical anastomosis of free omental grafts: experimental animal model of a new operative technique in dogs. AB - Our objective was the elaboration of a new animal model for the free transplantation of an omental flap and the examination of its viability in dogs. The cooled omental flap from the abdomen was freely transplanted to the lateral cervical region, and its blood supply was established with microsurgical anastomoses. The technique was developed in 5 dogs, and short-term survival examinations were later carried out in 3 cases by means of this method. Postoperative viability was assessed by angiography, methylene blue testing, and histology. Of the 3 transplanted grafts, 2 still survived 1 week after the operation. For technical reasons, 1 flap thrombotized. For determination of the viability of the transplanted graft, histology proved best. Vital reactions, including granulation tissue and angiogenesis, were present on the histological slides. The short-term survival of an omental flap can be ensured with microsurgical transplantation in dogs. PMID- 14557994 TI - Changes in microcirculation after ischemic process in rat skeletal muscle. AB - Laser Doppler flowmetry (LDF) is a good method to investigate tissue microcirculation, but it has many standardization and measuring problems. To exclude these effects, we performed a test using LDF on rat skeletal muscle. In 12 CD outbred anesthetized rats, bilateral femoral vessels and the quadriceps femoris muscle were exposed. The left femoral artery and vein were clamped for 1 h by microvascular clips (ischemic side). On the right side, no other intervention was made (control side). An LDF probe was applied on the medial vastus muscle. Short-term occlusions (2-3 s) were performed before and after the 1-h clamping and on the control side while LDF curves were registered. The halftimes of ascending curves on the ischemic side were significantly elongated vs. the condition before clamping (P = 0.0007) or the control side (P = 0.0017). Microcirculatory changes caused by 1-h ischemia were shown by this simple, quick, and well-reproducible test on rat skeletal muscle. PMID- 14557995 TI - Effect of partial hepatectomy on in vivo engraftment after intravenous administration of human adipose tissue stromal cells in mouse. AB - Their multidifferentiation potential makes human mesenchymal stem cells (hMSCs) candidates for cell-based therapeutic strategies for tissue injuries and for hematopoietic disorders by both local and systemic application. Despite their potential clinical utility in cellular and gene therapy, the fate of adipose stromal cells (ATSCs) after systemic administration is mostly unknown. In this study, we investigated the distribution of ATSCs injected intravenously and the effect of partial hepatectomy on their distribution. Adipose tissue stromal cells (hATSCs) were obtained from adipose tissues of adult human donors. Under appropriate culture conditions, hATSCs were induced to differentiate into osteocytes and adipocyte lineages. The hATSCs were marked by infection with the lacZ-adeno virus, and the distribution of injected cells was examined by X-Gal staining. Immunosuppression was achieved by the administration of cyclosporin into mice. The hATSCs were engrafted onto various tissues, including brain, thymus, heart, liver, and lung, after intravenous administration. Liver regeneration induced by partial hepatectomy enhanced the integration of hATCSCs into the liver. These results demonstrate that hATSCs have the ability to proliferate extensively in culture, and that they maintain their multilineage differentiation potential in vitro, establishing their progenitor cell nature. These cells are promising candidates for developing novel cell-based therapeutic approaches to postnatal tissue repair. PMID- 14557996 TI - Non-viral human IL-10 gene expression reduces acute rejection in heterotopic auxiliary liver transplantation in rats. AB - We studied nonviral delivery, expression, and the effect of the human interleukin 10 (Hu IL-10) gene on the rat model of heterotopic auxiliary liver transplantation (HALT). Two previous pilot studies showed remarkable expression of the Hu IL-10 gene in donor and recipient rats, and a decreasing effect of acute rejection in certain cases. In this study, we focused on the efficacy of Hu IL-10 gene expression to decrease acute rejection compared with cyclosporine A (CyA) in a HALT model. Three study groups and one control group were designed. Each group consisted of 6 DA donor and 6 Lewis recipient rats, which underwent HALT. In the control group, donors and recipients were not treated at all. In group II, recipients were treated with one dose of CyA. In group III, donors were treated with Hu IL-10 plasmid. In group IV, donors were treated with Hu IL-10 plasmid, and recipients were treated with one dose of CyA. Rejection was established by histopathology: it revealed 100% rejection in control and 33.3% rejection in study groups II, III, and IV. Human IL-10 gene expression prevented acute rejection with the same efficacy as CyA in the HALT model in rats. PMID- 14557997 TI - Vasectomy reversal and spermatic granuloma: experimental investigation. AB - The aim of our study was to investigate the healing process after vasectomy reversal, comparing two suturing techniques. Eighty Wistar rats received a two layer vaso-vasostomy on the right side, and 40 of them also received a single layer vaso-vasostomy on the left side. Twenty rats each time were sacrificed at 30, 45, 60, and 90 days. Results were evaluated in order to compare the two techniques used and to describe the progression of healing, with special reference to anastomotic patency rate, histologic aspect of the epididymus and testicle, and the presence and size of spermatic granuloma. We observed 45% patency with the double-layer technique compared to 20% with the single-layer technique. The testicle appeared atrophic in 20% of double-layer cases compared to 40% of single-layer cases. Comparison over time showed a progressive increase in patency of the anastomosis, reduction of spermatic granuloma, and regression of testicular damage. Our results show the superiority of the double-layer technique, and contribute to an understanding of the role of two different suturing techniques in the outcome of vasectomy reversal. PMID- 14557999 TI - Needles to promote ventricular blood into the ischemic myocardium applied in a rat heart transplant model: an acute observation [corrected]. AB - Two needles were designed in order to revascularize an ischemic myocardium in the event of left coronary artery occlusion. This study was conducted by performing the Lee modified Fox-Montorsi heart-lung transplant on 25 San Diego Microsurgical Institute-bred Sprague Dawley rats that were subjected to left coronary artery ligation in each case. Of these 25 rats, a straight-porous (SP) needling procedure was applied to 9 heterotopically transplanted rat hearts, and a distinct horseshoe (HS)-shaped needle application was performed on the remaining 16 heterotopically transplanted rat hearts. This report represents an acute study on the efficiency of these two needles to transmit oxygen-rich blood from the left ventricle into the ischemic myocardium. Doppler readings for male vs. female transplants showed that the control peak (PK) and mean (MN) kHz values are on the average of 0.20 kHz higher in males than in females. However, control heart rate values in both sexes are approximately equal. Ligation of the left coronary artery caused a dramatic decrease of PK and MN kHz values in both sexes, while heart rate showed no significant decrease from the original control values in response to ischemia. Application of the SP needle showed only a slight return of PK and MN values in both sexes, but heart rate values increased to levels higher than the original control values. The HS needling procedure was able to recover approximately 80% of the control PK and MN kHz values in both sexes. Thus, these data indicate that the HS needle can successfully transmit left ventricular blood into the myocardium. PMID- 14557998 TI - Consecutive en-bloc liver (30%)-pancreas-duodenum-spleen-stomach transplant in Lewis rats. AB - It is well-known that 30% of the remaining liver mass, following partial hepatectomy, regenerates to full original mass within 2 weeks in rats. In order to carry the transplanted rat liver to repeated transplantation, a technique of combining 30% of the liver with the pancreaticoduodenum and spleen transplantation is performed in this consecutive organ transplantation study. Our laboratory observed several 37-month-old transplanted rats by carrying through 2 3 generations, and histological disclosure were made. Because the partial liver transplants did not regenerate after the transplantation with other splanchnic organs, this technique is not so difficult though subsequent surgical maneuvers are needed and the liver histology proved entirely normal in every aspect when followed beyond the rat's life span of 24 months. PMID- 14558000 TI - Observations of rat ovarian-splenic consecutive transplants. AB - We examined the results after implantation of ovarian follicles by various modes in a total of 82 cases. One or five ovarian follicles were implanted into spleens in castrated female rats. In 20 cases among these, each five follicles were implanted into native and transplanted spleens after spleen transplantation (double implantation of the ovary). Through consecutive spleen transplantation, we observed the results beyond the rat's life span for a limited period. In many cases, we found a more aggressive form of malignant tumor, i.e., dysgerminoma. We present the results and discuss the modes of implantations and possible pathogenetic mechanisms of tumor formation. PMID- 14558001 TI - Effect of conversion from cyclosporine A to mizoribine on transplant arteriosclerosis in rat aortic allograft models. AB - Recent studies suggested that conversion from cyclosporine A (CsA) to antimetabolic agents protects allograft vasculopathy. However, the mechanism of this beneficial effect by the conversion therapy is not fully understood. In the present study, we investigated the effects of conversion from CsA to antimetabolic agent mizoribine (MZR) on the formation of transplant arteriosclerosis in rat aortic allografts. Conversion from CsA to MZR significantly prevented intimal hyperplasia and perivascular inflammatory cell infiltration at 28 days after aortic transplantation. These findings suggest that conversion therapy from CsA to antimetabolic agents might have therapeutic potential in transplant patients with chronic vascular rejection. PMID- 14558002 TI - Ischemia and hepatic reperfusion: is it possible to reduce hepatic alterations? AB - Our aim was to evaluate liver damage after ischemia and reperfusion, and at the same time test the effectiveness of some drugs in preventing these alterations. For this study, we utilized 50 rats divided into four groups: three underwent hepatic ischemia through occlusion of the portal vein and hepatic artery for 30 min, and one underwent a sham operation. In all groups, hepatic enzymes and bilirubine were tested at 2 h, 3 h, 4 h, 24 h, and 30 h. The drugs utilized were: L-arginine, donor of nitric oxide, and L-canavanine, inhibitor of nitric oxide synthase (NOS). Our data showed that the drugs tested could make an improvement in hepatic function after ischemia/reperfusion, preventing its damage. These preliminary results could suggest a clinical application in order to prolong ischemic period during liver transplantation or liver resection in cirrhotic patients. PMID- 14558003 TI - Microsurgical training course for clinicians and scientists at a German University hospital: a 10-year experience. AB - Microsurgical techniques are being increasingly applied in almost all surgical disciplines. However, the opportunities to learn these skills in a structured course are rare. We have conducted a 5-day microsurgical training course on a yearly basis since 1991. The course follows step-by-step training, starting with nonvital models for vascular and nerval microanastomoses. As the participants improve, exercises on laboratory animals are offered to close the gap between nonliving models and the clinical situation. Lectures provide theoretical and clinical background information. Clinical and experimental applications can be witnessed and practiced in a second part of the course. With this step-by-step curriculum, we conduct a successful training program, e.g., each participant is able to perform microvascular and nerval anastomoses on a reproducible basis. The organization, program, and execution of the training course are presented, together with an evaluation of the course concept by the participants concerning expectations, learning success, and level of satisfaction. PMID- 14558005 TI - Rat liver transplantation for total vascular reconstruction, using a suture method. AB - We developed a novel protocol for rat orthotopic liver transplantation (OLT), using a suture method to establish hepatic artery flow. After determining that early inferior vena cava (IVC) unclamping maintained better circulation compared with the portal vein (PV) using porto-systemic shunted recipients, we developed a rat OLT model with total vascular reconstruction using a suture method. After connecting the suprahepatic IVC, the infrahepatic IVC was anastomosed, using a running suture method. IVC circulation was established immediately. The PV was anastomosed without intestinal congestion, using porto-systemic shunted recipients. The aortic conduit, including the donor celiac and hepatic artery, was anastomosed to the recipient abdominal aorta end-to-side. Eight of 11 OLT cases (72.7%) survived indefinitely. Biliary connection was achieved using a one stent method. Three cases died 3-5 days postoperatively. Hepatic angiography showed good patency. The graft liver was histologically normal in long-surviving rats. PMID- 14558004 TI - Multiorgan transplantation with a new organ-chip technique in mice: preliminary histological data. AB - A simple model was developed for multiorgan liver-kidney-spleen-intestine transplantation on 108 inbred mice. Donor operations included hepatectomy, nephrectomy, splenectomy, and jejunum segment resection. Following removal of the organ, small slices or abdominal organ "chips" were prepared. During multiorgan recipient operations, chips from each of these organs were transplanted into the omentum; in the control single-organ groups, only 1 organ was transplanted. All animals survived. Biopsies were taken for histology after 6 weeks. All organs were found to have developed a blood supply. In the liver chips, hypertrophied cells could be detected. In the margin of the kidney tissue, both the glomeruli and tubules were preserved. Lymphoid zone and red pulp were intact in spleen chips. All layers of the intestinal chips were identifiable and contained intraluminal mucinous substances. This model is a simple surgical intervention with the possibility of the investigation of 4 organs. PMID- 14558006 TI - Synergistic effects of RAD and Neoral in inhibition of host-vs.-graft and graft vs.-host immune responses in rat small-bowel transplantation. AB - The combined effects of RAD and Neoral were tested in a rat orthotopic small bowel transplantation model. Seven groups (n = 6) were involved in this study, and each one was included in three rejection models for the evaluation of host vs.-graft disease (HVG) (LBN-F1 to LEW), graft-vs.-host disease (GVH) (LEW to LBN F1), and combined HVG and GVH immune responses (BN to LEW). Both drugs were administered orally throughout the study. Low doses of RAD (1.0-2.5 mg/kg/day) combined with Neoral (2.0-5.0 mg/kg/day) produced strong synergistic effects in the prolongation of small-bowel graft survival in HVG (combination index, CI = 0.095, 0.1212), GVH (CI = 0.027, 0.020), and combined HVG and GVH immune responses (CI = 0.070, 0.301). The combination therapy of RAD and Neoral produces a strong synergistic effect toward the inhibition of HVG, GVH, and combined HVG and GVH immune responses in a rat small-bowel transplantation model. PMID- 14558007 TI - Hematological, hemorheological, immunological, and morphological studies of spleen autotransplantation in mice: preliminary results. AB - Using a spleen autotransplantation model, we conducted hematological, hemorheological, immunological, and morphological studies in mice 6 weeks after splenectomy. Sixty male and female A/J inbred mice were equally divided into 3 groups: 1) SE group, splenectomy was performed; 2) AU group, spleen chips were autotransplanted into the omentum without vascular anastomosis following splenectomy; and 3) C group (controls), no intervention in these mice. At postoperative week 6, the following studies were performed: 1) measurement of hematological parameters; 2) hemorheological studies, including relative cell transit time (RCTT) and fibrinogen levels; and 3) activity of peripheral phagocytes, measured by zymozan-induced chemiluminescence, which was calculated in stimulation index values (SI). In addition, histological investigations of autotransplants were conducted. Erythrocyte mean cell volume and platelet counts, RCTT, fibrinogen levels, and activity of phagocytes were significantly higher in the SE group, compared to those in the C group. In the AU group, these parameters were similar to those in the C group. Morphologically, the transplanted spleen showed normal histology. These data indicate that the transplanted spleens restored their function. We conclude that spleen autotransplantation reserves the normal morphology of spleen and restores most of the spleen's hematological, hemorheological, and immunological functions. Both SI index and erythrocyte deformability can be an informative detection of decreasing splenic function. These data suggest that spleen autotransplantation may provide a useful tool to prevent complications following splenectomy in a clinical setting. PMID- 14558008 TI - Prolonged survival of heart allografts transduced with AAV-CTLA4Ig. AB - Organ grafts transduced with gene-encoding immunosuppressive molecules are a less toxic approach to preventing graft rejection. Adenovirus vectors have been widely tested with unsatisfactory results, while adeno-associated virus (AAV) is smaller and elicits a low host humoral response. We constructed an AAV vector containing the mouse CTLA4Ig gene. B10 (H2(b)) cardiac grafts were transduced with AAV CTLA4Ig by coronary infusion. AAV-LacZ vectors were used as reporters and controls, and the expression of beta-gal was determined by X-gal staining. Thirty percent to 40% of myocytes displayed strongly positive X-gal staining after infusion with AAV-LacZ. Additional infusion with vascular dilator reagents did not improve the transduction rate. Survival of B10 heart allografts transduced with AAV-CTLA4-Ig was significantly prolonged in C3H (H2(k)) recipients. These data demonstrate that AAV vectors can efficiently be transduced into the mouse myocardium by coronary infusion. Graft transduction with AAV-CTLA4Ig may be a novel approach to preventing allograft rejection. PMID- 14558009 TI - Administration of antisense oligodeoxyribonucleotides targeting NF-kappaB prolongs allograft survival via suppression of cytotoxicity. AB - Presentation of alloantigens by antigen-presenting cells (APC) deficient in expression of costimulatory molecules (CM) induces alloantigen-specific hyporesponsiveness or tolerance. NF-kappaB has been shown to be a crucial transcription factor that regulates CM expression on the surface of APC. We designed an antisense phosphorothioate oligodeoxynucleotide (ODN) to specifically inhibit mRNA of mouse NF-kappaB, resulting in a deficiency in CM expression on APC that may prolong organ allograft survival. Anti-NF-kappaB ODN was delivered systemically by an osmotic pump implanted in the abdominal cavity of recipients following transplantation of vascularized heart allografts. The animals in control groups were given scrambled control ODN or were left untreated. Normal C3H (H2(k)) recipients rejected B10 (H2(b)) heart allografts at a median survival time (MST) of 15 days. A 14-day administration of 12.5 mg/kg/day anti-NF-kappaB ODN prolonged the survival of cardiac allografts to an MST of 25 days (P <0.05). In contrast, a scrambled control ODN was not effective (MST 17 days, P >0.05 compared to untreated controls). To investigate the underlying mechanisms of the immunosuppressive effect of anti-NF-kappaB ODN administration, graft infiltrating cells, spleen cells, and serum were collected from animals on day 7 posttransplant. Freshly isolated graft-infiltrating cells from anti-NF-kappaB ODN treated recipients exhibited significantly decreased donor-specific CTL activity. Generation of CTL activity of spleen T cells from anti-NF-kappaB ODN-treated recipients was also impaired compared with untreated animals. Administration of anti-NF-kappaB ODN did not influence the titers of complement-dependent cytotoxic antibodies. These data suggest that treatment with anti-NF-kappaB ODN markedly inhibits the cellular response of allograft recipients, resulting in significant prolongation of allograft survival. Antisense ODN therapy targeting NF-kappacB may be a novel strategy for future immunosuppressive therapy. PMID- 14558010 TI - Comparison of in vivo lymphocyte proliferation between allogeneic and xenogeneic heart transplantation in mice. AB - There are controversial in vitro data comparing the strength of the cellular immune response between allogeneic and xenogeneic stimulator/responder combinations. The present study therefore compares in vivo lymphocyte proliferation using heart transplantation (HTx) models in mice. Heterotopic HTx into BALB/c mice was performed using donor organs from mice (BALB/c and C57BL/6) or Lewis rats. Intraperitoneally given bromodeoxyuridine (BrdU) was incorporated into the DNA and was subsequently analyzed by flow cytometry. On postoperative days 3 and 5, proliferation of splenocytes, CD4(+) T-lymphocytes, and CD19(+) B lymphocytes was significantly higher after xenogeneic than after allogeneic and isogeneic HTx. No significant difference was observed when proliferation of CD8(+) lymphocytes was determined. The increased in vivo proliferation after xenotransplantation may reflect an earlier and probably stronger cellular immune response compared to allogeneic transplantation. The higher CD4(+) lymphocyte proliferation underscores the importance of this cell population in xenograft rejection. PMID- 14558011 TI - Short-term immunosuppression after rat parathyroid allotransplantation. AB - The aim of this study was to evaluate whether short-term postoperative immunosuppression is able to sufficiently prolong graft survival after experimental allogeneic parathyroid transplantation. Heterotopic parathyroid transplantation was performed in 6 groups: 1) syngeneic control Lewis (LEW) to LEW; 2) allogeneic control Wistar-Furth (WF) to LEW; 3-5) WF to LEW plus short term immunosuppression, postoperative days 1-13 (cyclosporine 5/10/20 mg/kg); and 6) WF to LEW plus 10 mg/kg CyA from preoperative day 7 to postoperative day 7. Graft function was examined up to 60 days; histological and immunohistological examination was performed on all grafts with impaired function. Graft function after syngeneic transplantation was indefinite, while recipients of allogeneic grafts turned hypocalcemic after 13 +/- 2 days. With immunosuppression, graft function was 21 +/- 2 days (groups 5 and 6) and 28 +/- 3 days (groups 3 and 4). Histologically, a cellular infiltrate responsible for graft destruction was found. The results show that indefinite parathyroid allograft survival cannot be achieved by short-term immunosuppression alone. Whether the combination of an additional graft pretreatment and immunosuppression has an impact on graft function will be further examined. PMID- 14558012 TI - Optimization of a metastasizing human gastric cancer model in nude mice. AB - Our purpose was to optimize the surgical orthotopic implantation (SOI) technique to create a reproducible gastric cancer model in nude mice with stable tumor growth and metastasizing course. We performed xenotransplantation of primary human tumor specimens from patients with gastric cancer (series 1) and orthotopic transplantation of tumor specimens originating from the gastric cancer cell line 23132/87 (series 2). All specimens were transplanted using microsurgical techniques. The two series were compared with regard to tumor growth rates and kinetics, development of metastases, and induction of minimal residual disease (MRD), as determined by histology and PCR techniques. In series 1 mice, the tumor growth rate was slow; in series 2 mice, it was both fast and reproducible. Unlike animals in series 1, animals in series 2 developed metastases and MRD. In conclusion, the optimized SOI technique presented here represents a reproducible and reliably metastasizing gastric cancer model. PMID- 14558013 TI - Progressive gastric perfusion in rats: role of ischemic conditioning. AB - Occult ischemia of the mobilized stomach is usually related to the dehiscence of an esophagogastric anastomosis. The principle of ischemic conditioning was studied to verify its clinic use. This study aims to evaluate progressively the tissue perfusion of the stomach in ischemic conditioning, establishing the best moment for gastric transposition. Twenty-four male EPM-1 Wistar rats were used, which also underwent partial desvacularization of the stomach by ligature of the left gastric vessels. Tissue perfusion was measured through flowmetry by laser Doppler (tissue perfusion unit; TPU) in the antrum (10 mm distal from the cardiac region). This measurement was done before (baseline) and immediately after the ligature, and on different postoperative days (POD) (days 3, 7, 10, and 14). A statistical analysis was done with nonparametric tests (P or =37 at delivery), these would have been respectively 40%, 7% and 1 : 96 between 1990 and 1994, and 40%, 9% and 1 : 107 between 1995 and 1999. CONCLUSIONS: Ongoing audit of DS prenatal programmes could be derived from computerised maternity data sets if they included fetal deaths, and relevant laboratory and ultrasound findings. PMID- 14558025 TI - Craniofacial dysmorphology and three-dimensional ultrasound: a prospective study on practicability for prenatal diagnosis. AB - OBJECTIVE: The aim of this study is to compare two-dimensional and three dimensional ultrasound for the visualization and diagnosis of craniofacial dysmorphism. METHODS: In this prospective study, we performed three-dimensional (3D) ultrasound following good-quality two-dimensional (2D) ultrasound in an at risk population. Findings from 2D and 3D examination were noted. RESULTS: Our series included 41 patients. In 20 cases, 3D performed better than 2D, and in two cases 2D was more informative. In 9 cases, there was no difference between the performance of the two, and in two cases both 2D and 3D gave inadequate results. However, in 7 cases 3D images could not be obtained. CONCLUSION: Although it is rarely decisive, 3D ultrasound is of interest when it comes to the precise description of craniofacial dysmorphisms and the study of the fetal ears. PMID- 14558026 TI - Cryptic 1p36.3/6q25.2 translocation in three generations ascertained through a foetus with IUGR and cerebral malformations. AB - Here we describe a foetus with intrauterine growth retardation (IUGR), cerebral malformations and a 46,XY,der(1),t(1;6)(p36.3;q25.2) karyotype owing to a familial cryptic translocation segregating in three generations. A balanced translocation was present in the mother, the maternal uncle, the aunt and the grandmother. A female first cousin with dysmorphisms, hydrocephalus and mental retardation was a carrier of a partial trisomy 1p and a partial monosomy 6q. Multiple miscarriages were present in the family pedigree. Parents of the foetus had three other pregnancies: a male with a balanced translocation, and two foetuses with 1p36.3-pter monosomy and 6q25.2-qter trisomy. PMID- 14558027 TI - Mild pyelectasis: evaluating the relationship between gestational age and renal pelvic anterior-posterior diameter. AB - OBJECTIVE: To determine the relationship between gestational age and renal pelvic anterior-posterior diameter and the feasibility of developing gestational age specific thresholds for the diagnosis of mild pyelectasis. METHODS: Cross sectional study of 420 singleton fetuses between 16 and 39 weeks' gestation. The mean renal AP diameter as a function of gestational age was determined using fractional polynomial regression models and centile plots were generated. Assessment of goodness of fit for each regression model was performed. RESULTS: There was a positive correlation between gestational age and renal pelvic AP diameter (Pearson's Correlation Coefficient 0.65). Using the derived mean and standard deviations of renal AP diameter, gestational-age specific 95% reference levels were generated. The sensitivity, specificity, positive, and negative predictive values of using the gestational age-specific cutoffs for predicting persistent postnatal renal anomaly were 80%; 99%; 29%; and 99% respectively. CONCLUSION: There is a positive correlation between gestational age and renal pelvic AP diameters. Reliable gestational age-specific renal AP thresholds for diagnosis of pyelectasis are provided. PMID- 14558028 TI - Do women prefer to have screening tests for Down syndrome that have the lowest screen-positive rate or the highest detection rate? AB - OBJECTIVE: To assess whether women have a preference for Down syndrome screening test performance. METHODS: A structured questionnaire exploring women's preferences for screening test performance was administered to women attending their first prenatal visit who wished to have Down syndrome screening performed. RESULTS: One hundred and twenty women were interviewed. The majority of women (n=80) chose a screening test with a low screen-positive rate rather than the highest detection rate. The reasons given for this preference were a desire to minimise the risk of miscarriage of a normal baby and a belief that a detection rate of 80 to 90% was acceptable. However, older women (>37 years) chose a test with the highest detection rate possible, regardless of the higher screen positive rate, preferring to miscarry a normal baby as a result of a diagnostic test rather than miss the detection of a baby with Down syndrome. Preferences were not influenced by previous screening experience. CONCLUSIONS: Women express different preferences for screening test performance. Maternal age rather than previous screening experiences appears to be the major influence in these choices. PMID- 14558029 TI - First-trimester screening for Down syndrome in France combining fetal nuchal translucency measurement and biochemical markers. AB - BACKGROUND: In France, there is a strictly regulated National Screening Programme for Down syndrome, based on second-trimester maternal serum markers. A prospective study of nuchal translucency together with retrospective evaluation of maternal serum markers was carried out to inform decisions on whether to move the programme to the first trimester. METHODS: Between January 1998 and June 2001, all women who presented for their prenatal care at 12 participating maternity units were, regardless of age, invited to provide a blood sample and to attend for an NT scan at 11 to 13 weeks. The results were used to derive Gaussian distribution parameters. Detection and false-positive rates were computed in two ways: statistical modelling and directly. The cut-off risk was 1 in 250 at term. RESULTS: A total of 5694 women with singleton pregnancies were screened including 26 with Down syndrome and 24 with other aneuploidies. The model-predicted detection and false-positive rates for combined ultrasound and serum screening were 81 and 4.5% compared to 64 and 6.0% for ultrasound alone. The directly observed rates were 73 and 4.7%, compared to 62 and 5.0% respectively. CONCLUSION: In France, first-trimester screening with nuchal translucency and maternal serum markers is likely to achieve a high screening efficiency. This has important implications for the national screening policy. PMID- 14558030 TI - Fetal cells and DNA in maternal blood. AB - Although fetal cells have been known to escape to the maternal circulation for a number of years, research attempts to use them for prenatal diagnosis have not had any consistent success. This review attempts to trace the history of such attempts and to document their progress and reasons for success or failure. The opinions of recent conferences including that of the US National Institute of Child Health and Human Development, a sponsor of major US research in the field, are reported and discussed. It is concluded that although basic work has demonstrated the biologic availability of both fetal cells and their free DNA representatives in the maternal circulation at gestational ages relevant to prenatal diagnosis, much work remains to develop practical technology for their consistent recovery and assay. PMID- 14558031 TI - Fetal chromosomal analysis of pregnancies following intracytoplasmic sperm injection with amniotic tissue culture. AB - OBJECTIVES: The objective of this study was to determine the incidence of chromosomal anomalies in a complete cohort of ICSI pregnancies. METHODS: From January 1996 to December 2000, 1500 consecutive patients who had become pregnant after Intracytoplasmic Sperm Injection (ICSI) were given prenatal genetic counseling and 98 of them (6.5%) who accepted amniocentesis were studied. Amniocentesis was performed between 14 and 20th weeks of their pregnancy. Amniotic tissue cultures were processed according to Hoehn et al. A minimum of 20 metaphases were examined on each preparation. Both parents were also evaluated with peripheral blood karyotyping. RESULTS: Amniocentesis and karyotyping was performed to evaluate the 142 fetuses from 98 ICSI pregnancies. Karyotypes from peripheral leucocytes of the parents were also evaluated. Chromosomal anomalies were detected from 6 out of the 142 (4.2%) fetuses. The anomalies were as follows, '46,XX/69,XXX/92,XXXX', '46,XY/69,XXY/92,XXYY', '47,XY + 21', '46,XY/47,XY + 7', '47,XXY', and '45,X0'. All except one pregnancy were terminated with the consent of the couples. Fetal tissue cultures were also studied after the termination of the pregnancy in order to confirm prenatal diagnosis. We did not detect abnormal karyotype from the parents of these fetuses. CONCLUSIONS: The ratio of chromosomal abnormalities seems to be slightly increased in ICSI pregnancies. Paternal factors due to morphologically abnormal spermatozoa, maternal factors due to increased mother age and more frequently, de novo occurring sex chromosomal abnormalities may be responsible for this outcome. PMID- 14558032 TI - Maternal weight correction of maternal serum PAPP-A and free beta-hCG MoM when screening for trisomy 21 in the first trimester of pregnancy. AB - OBJECTIVE: To assess the suitability of either the log-linear or reciprocal linear regression procedure for maternal weight correction of biochemical marker MoMs in the first trimester. METHODS: Data from two prospective first-trimester OSCAR screening programmes including 32,010 women with first-trimester maternal serum-free beta-hCG and PAPP-A measured by the Kryptor analyser was analysed by regression analysis to provide parameters for the log-linear and reciprocal linear MoM correction procedures. Assessment was made by goodness of fit to the data. The impact on detection rate and false-positive rate of the different correction procedures was assessed using statistical modelling with biochemical markers alone. RESULTS: Both log-linear and reciprocal-linear correction were shown to fit the data well. For free beta-hCG, the log-linear procedure was marginally superior to the reciprocal-linear procedure (r2=0.986 v 0.980), whilst for PAPP-A the reciprocal-linear procedure was marginally better (r2=0.991 v 0.985). Log-linear correction reduced the variance for both markers more than did the reciprocal-linear procedure. For free beta-hCG, the sd was reduced from 0.2675 to 0.2605 and for PAPP-A, it was reduced from 0.2545 to 0.2336. Correcting for maternal weight was shown to reduce the population false-positive rate from 7.0 to 6.5%, whilst maintaining the same detection rate at a risk cut-off of 1 in a 100. At individual levels, a two-fold variation in risk was demonstrated depending upon the individual's weight. CONCLUSIONS: To provide accurate individual patient-specific risks for trisomy 21, maternal weight must be taken into account and should be a mandatory data item for screening programmes. Maternal weight correction in the first trimester using free beta-hCG and PAPP-A can be best achieved using the log-linear procedure. PMID- 14558033 TI - Trisomy 13 and trisomy 18 in a defined population: epidemiological, genetic and prenatal observations. AB - OBJECTIVES: To establish precise incidence figures for trisomy 13 and trisomy 18 in the former Trent region, to identify current prenatal diagnostic practice, and to assess the potential impact of the introduction of recently devised prenatal diagnostic practices. METHODS: An audit of all cases of trisomy 13 and trisomy 18 ascertained through the records of the Trent Congenital Anomalies Register and the Trent Regional Cytogenetic Laboratories. RESULTS: Forty-four cases of trisomy 13 and 88 cases of trisomy 18 were ascertained. Advanced maternal age effects were observed. Of all cases, 64% were first detected through chromosomal analysis initiated because of abnormalities noted on fetal anomaly scanning in the second trimester, whereas only 3% of cases were detected through the serum-screening programme currently offered for Down syndrome. In 11% of cases, the diagnosis was first suspected after birth. Twelve percent of couples chose to continue pregnancy following chromosomal confirmation of a suspected diagnosis. CONCLUSION: The introduction of a highly sensitive prenatal diagnostic screening programme would have a major impact on the timing and proportions of all trisomy 13 and 18 cases diagnosed in pregnancy as gauged by current practice. It is important that health professionals involved in prenatal counselling be aware that, as with Down syndrome and anencephaly, around 12% of prospective parents of a child with trisomy 13 or 18 choose to continue rather than terminate the pregnancy. PMID- 14558034 TI - Certification of competence in performing specific procedures or tests in screening practice. PMID- 14558035 TI - Molecular diagnosis in a pregnancy at risk for both spondyloepiphyseal dysplasia congenita and achondroplasia. PMID- 14558036 TI - Low first-trimester pregnancy-associated plasma protein-A and Cornelia de Lange syndrome. PMID- 14558037 TI - Reversed diastolic flow in the middle cerebral artery: its clinical value in fetal growth restriction. PMID- 14558038 TI - Prenatal findings for complete trisomy 9. PMID- 14558039 TI - Maternal experience influences the establishment of visual/auditory, but not olfactory recognition of the newborn lamb by ewes at parturition. AB - Maternal behavior in sheep is characterized by the rapid establishment of individual recognition of the lamb through the use of different sensory modalities. Olfactory recognition mediates acceptance at suckling whereas visual/auditory cues are involved in recognition from a distance. This study investigates (a) the timing of both types of recognition and (b) whether they can be influenced by maternal experience. Olfactory recognition was assessed at lambing, 30 min, 1, 2, or 4 hr postpartum by presenting successively an alien and the familiar lamb. Recognition at a distance was assessed at 6, 8, and 12 hr postpartum by using a two-choice test between the familiar and an alien lamb. A majority (33/51) of ewes showed selectivity at suckling as early as 30 min after parturition, and no differences due to maternal experience were observed. By contrast, in the two-choice recognition test, multiparous ewes showed a preference for their familiar lamb at 6 hr whereas primiparous mothers did so only after 24 hr of mother-young contact: The performance of biparous mothers was intermediate. Thus, both types of recognition can be rapidly established after parturition, and maternal experience has a differential effect on the dynamics of these learning processes. PMID- 14558040 TI - Complete maternal deprivation affects social, but not spatial, learning in adult rats. AB - The effects of maternal deprivation on learning of social and spatial tasks were investigated in female adult rats. Pups were reared artificially and received "lickinglike" tactile stimulation (AR animals) or were reared with their mothers (MR animals). In adulthood, subjects were tested on paradigms of spatial learning and on paradigms involving learning of social cues. Results showed that maternal deprivation did not affect performance on spatial learning, but it did impair performance on the three social learning tasks. The AR animals made no distinction between a new and a previously presented juvenile conspecific. AR animals also responded less rapidly than MR animals at test for maternal behavior 2 weeks after a postpartum experience with pups. Finally, AR animals did not develop a preference for a food previously eaten by a familiar conspecific whereas MR animals did. This study indicates that animals reared without mother and siblings show no deficits in spatial tasks while showing consistent deficits in learning involving social interactions. PMID- 14558041 TI - Arousal, but not nursing, is necessary to elicit a decreased fear reaction toward humans in rabbit (Oryctolagus cuniculus) pups. AB - Rabbits that are handled at the time of feeding during the first week postpartum show reduced fear of humans later in their lives as compared to unhandled controls. Effective handling has been shown to be confined to a sensitive period. Our study aimed to investigate if feeding itself (provided by a second doe, 6 hr after the standard nursing time) affects the levels of fear of humans later in life. Our results showed that (a) the prenursing state of excitement is only characteristic of the standard nursing and is not elicited by a second feeding 6 hr past the usual nursing time, repeated daily across the first week postpartum; and (b) handling linked to a second feeding 6 hr after the standard nursing does not reduce fear responses toward humans at weaning. We conclude that the aroused state, per se, is essential for the reduction of a rabbit's fear response toward humans provoked by early handling. PMID- 14558042 TI - Alcohol intake in social housing and in isolation before puberty and its effects on voluntary alcohol consumption in adulthood. AB - We assessed the effects of alcohol exposure in social and isolation housing before puberty on the alcohol consumption later in adulthood. From 25 to 35 days of age, Wistar male rats were exposed to either (a) continuous isolation; (b) continuous social housing; (c) continuous isolation, but placed in social housing during 12 hr every other day; or (d) continuous social housing, but placed in isolation during 12 hr every other day (SOIS group). All males were exposed to 8% ethanol as the only available liquid 12 hr/day every other day in this prepubescent period and to continuous free-choice access to 8% ethanol or water in both social and isolation housing when adulthood was reached. Alcohol consumption before puberty was higher in the group permanently housed in isolation. Both voluntary alcohol intake and preference for alcohol in adulthood also were higher during the isolation than in the social condition in all groups, except in the SOIS group. Alcohol consumption was higher in the SOIS group than in the other groups only during social condition in adulthood. Food intake decreased during the social interaction of the groups that changed their condition from isolated to social or from social to isolated. Results support that (a) isolation facilitates alcohol consumption, and (b) rearing characteristics associated to alcohol intake at an early age can play an important role in later alcohol intake. PMID- 14558043 TI - Thermal benefit of sibling presence in the newborn rabbit. AB - Newborn rabbits compete vigorously for the mother's milk, and survivors benefit from littermate deaths. Here we report that rabbit pups also may benefit from littermate presence in terms of increased thermal efficiency. Pups nursed normally once a day by their mother but otherwise raised alone had a lower probability of survival, lower body temperature, and lower efficiency of converting milk into body mass than their siblings raised with littermates. The contribution of a more favorable thermal environment to the better growth and survival of group-raised pups was supported by the finding that single pups raised at higher ambient temperatures grew more rapidly than single pups raised at lower temperatures. These effects were most clearly seen across Days 2 to 5, after which time differences between treatment groups were no longer significant. Thus, the results of this study suggest that the benefits as well as costs of having siblings must be weighed against each other when considering the manner in which sibling presence influences individual development and survival. PMID- 14558044 TI - Retrieval difficulty and retention of reactivated memories over the first year of life. AB - A newly acquired memory is remembered longer when its retrieval is more difficult (i.e., when the time since the last retrieval is longer). In two experiments using an operant conditioning task with 198 infants between 3 and 12 months of age, we presently asked if a reactivated memory also is remembered longer when its retrieval is more difficult. During their first year, infants remember a memory reactivated 1 week after forgetting and the newly acquired memory equally as long. After determining the upper limit (UL) of reactivation at all ages (Experiment 1), we reactivated the memory near the UL and then measured its subsequent persistence (Experiment 2). We found that increasing the training reactivation interval enhanced retention at 6 months only. These data reveal that increasing retrieval difficulty affects infants' retention of newly acquired and reactivated memories differently. More generally, memory reactivation seems to disproportionately benefit retention in younger infants. PMID- 14558045 TI - Short-term stability of physiological measures in kindergarten children: respiratory sinus arrhythmia, heart period, and cortisol. AB - The short-term stability of measures of respiratory sinus arrhythmia (RSA), heart period, and salivary cortisol and the relation of these variables to one another and to measures of social competence in kindergartners were examined. Physiological measures were collected during multiple individual sessions with 30 kindergarten children, ages 5 to 6 years. Three sessions were scheduled at 2-week intervals and included a negative affect task as a challenge. Cardiovascular and cortisol responses to the challenge were assessed. Results indicated that baseline measures of RSA and heart period were stable across time. Baseline cortisol levels were not stable. The change in RSA in response to a task that elicited emotional responses was not stable over time. Consistent with findings observed in adults and older children, changes in RSA were related to the cortisol response. Children who increased cortisol over the course of Session 1 exhibited larger decreases in RSA than children who did not show increased cortisol levels. In addition, the children who had larger decreases in RSA had higher ratings of sociability on the Colorado Childhood Temperament Inventory. Greater decreases in heart period during the task were associated with lower ratings of shyness. The relation of RSA measures to ratings of social competence supports the Polyvagal Theory (Porges, 1995, 2001), which suggests that the ability to engage and disengage with objects and people in one's environment, necessary for social competence, is related to the regulation of vagal control of the heart. PMID- 14558046 TI - Rheumatology patients' use of complementary therapies: results from a one-year longitudinal study. AB - OBJECTIVE: To examine the natural history of complementary and alternative medicine (CAM) use and its impact on outcomes within a cohort of rheumatology patients. METHODS: Consecutive patients were recruited from 3 university and 3 private rheumatology practices. Baseline chart reviews provided demographic information and rheumatic diagnoses. Patients answered questions on CAM use and health status during 1 year. We identified correlates of 4 CAM usage patterns (started, maintained, stopped, nonuse) and compared outcomes among these groups. RESULTS: Of 232 baseline participants, 203 (87%) and 177 (76%) responded to the 6 and 12-month surveys. In each survey, approximately 34% reported currently using CAM. During the year, 44% of patients remained nonusers whereas 12% started, 22% maintained, and 22% stopped use. The most frequent reasons for stopping CAM were lack of effectiveness and expense. CAM users and nonusers had no difference in outcomes. CONCLUSIONS: Arthritis patients' usage behavior varied substantially, but CAM use was not associated with a difference in outcomes. PMID- 14558047 TI - Moberg picking-up test in patients with inflammatory joint diseases: a survey of suitability in comparison with button test and measures of disease activity. AB - OBJECTIVE: To assess and compare the suitability of Moberg pickup test (MPUT) and button test (BT) as indicators for functional impairment in patients with inflammatory joint diseases. METHODS: Measurements for 369 patients attending a rheumatology outpatient clinic were collected. In addition to MPUT and BT, measurements collected were grip strength, tender and swollen joint counts, visual analog scales for pain and disease activity, Health Assessment Questionnaire, C-reactive protein levels, and erythrocyte sedimentation rates. RESULTS: We found a significant relationship between MPUT and BT. Both tests show the same pattern of correlations with the other parameters, although all correlations are higher for MPUT. There is a significant sex and learning effect for the BT, which implies a confounding of hand function and motor abilities. A significantly higher proportion of patients was unable to complete BT. CONCLUSION: MPUT and BT measure comparable aspects of hand function. In several theoretical and practical aspects, MPUT seems superior to BT in arthritis. It is necessary to evaluate its value in long-term followup. PMID- 14558048 TI - Reactive hemophagocytic syndrome in adult systemic disease: report of twenty-six cases and literature review. AB - OBJECTIVE: To analyze specific clinical findings, underlying disorders, treatments, outcomes, and prognostic factors for reactive hemophagocytic syndrome (RHS) in systemic disease. METHODS: Data were collected using standardized forms as part of a French national survey. Adult cases without an underlying malignancy, diagnosed on bone marrow or lymph node biopsy, were included. RESULTS: Twenty-six cases (7 men, 19 women, mean age 47.4 +/- 17.7 years) were studied. Systemic diseases included systemic lupus erythematosus (n = 14), rheumatoid arthritis (n = 2), adult onset systemic Still's disease (n = 4), polyarteritis nodosa (n = 2), mixed connective tissue disease (n = 1), pulmonary sarcoidosis (n = 1), systemic sclerosis (n = 1), and Sjogren's syndrome (n = 1). RHS occurred in 2 distinct clinical settings in the course of systemic disease. RHS was associated with an active infection in 15 patients (bacterial infections, 10 cases; viral, 3 cases; tuberculosis, 1 case; and aspergillosis, 1 case) and with the onset of a systemic disease alone in 9 cases. Isolated RHS occurred in 2 cases. The overall mortality rate was 38.5%. Two factors were associated with mortality: corticosteroid treatment at the time of RHS diagnosis, and thrombocytopenia (odds ratio = 28, 95% confidence interval = 13.3-238.9). CONCLUSIONS: When RHS occurs in the course of an active systemic disease (situation only reported in cases of systemic lupus or adult Still's disease), immunosuppressive therapy should be used. In contrast, when RHS is present concomitantly with an active infection, immunosuppressive therapy needs to be lowered and antibiotic therapy should be instituted. PMID- 14558049 TI - Concordance of four methods of disability assessment using performance in the home as the criterion method. AB - OBJECTIVE: To determine the concordance of 4 methods of disability assessment with the criterion method. Performance testing in the home was selected as the criterion. METHODS: The task performance of 57 community-dwelling older women (>/=70 years) with knee osteoarthritis was examined through self report, proxy report, clinical judgment based on impairment measures, performance testing in an occupational therapy clinic, and performance testing in participants' homes. The 26 tasks represented 4 domains of daily living activities: 5 functional mobility, 3 personal care, 14 cognitively oriented instrumental activities of daily living (IADL), and 4 physically oriented IADL. RESULTS: In general, self reports and proxy reports had the highest concordance with in-home performance test results. Nonetheless, even for these methods, depending on task domain, the rate of discordance ranged from 31% to 54%, being least in personal care and greatest in the physically oriented IADL. CONCLUSION: Disability estimates based on self reports, proxy reports, clinical judgments, and hospital performance-based assessments are not interchangeable with in-home task performance. PMID- 14558050 TI - Osteoarthritis of the knee is associated with vertebral and nonvertebral fractures in the elderly: the Rotterdam Study. AB - OBJECTIVE: To study the association between prevalent radiographic osteoarthritis (ROA) of the knee and incident vertebral and nonvertebral fractures. METHODS: A sample of 2,773 subjects was drawn from the Rotterdam Study, a prospective population-based cohort study of the elderly. Status on knee ROA was assessed at baseline using the Kellgren score. Incident nonvertebral fractures were scored for all subjects, and for 1,466 subjects additional data on incident vertebral fractures were available. RESULTS: Although people with ROA had a higher bone mineral density (BMD), their incident fracture risk was increased as compared with those without ROA. After adjustment for potential confounding factors, including parameters of postural stability, the relative risks for incident vertebral and nonvertebral fractures in the presence of knee ROA were 2.0 (95% confidence interval [95% CI] 1.1-3.4) and 1.5 (95% CI 1.1-2.0), respectively. CONCLUSIONS: Knee ROA is associated with an increased risk of incident vertebral and nonvertebral fractures, independent of BMD and parameters of postural stability. PMID- 14558051 TI - Performance of a generic health-related quality of life measure in a clinic population with rheumatic disease. AB - OBJECTIVE: To assess the performance of a generic health-related quality of life (HRQOL) measure in a rheumatology clinic population. METHODS: Participants (n = 619) with fibromyalgia, rheumatoid arthritis, or osteoarthritis receiving care from rheumatologists completed mailed questionnaires that included the Behavioral Risk Factor Surveillance System (BRFSS) HRQOL measure and condition-specific measures assessing disability, pain, fatigue, and helplessness. The BRFSS assesses global health and number of days in the past 30 of poor physical or mental health or activity limitation. The overall sample was described, followed by comparison of adjusted scores on all HRQOL measures by diagnosis. RESULTS: Participants reported mild difficulty with activities of daily living, marked pain and fatigue, and moderate helplessness. Participants reported a mean of 8 or more days out of 30 of poor physical and mental health and activity limitations; more than 40% reported poor or fair health. Participants with fibromyalgia reported more ill health on condition-specific measures and the BRFSS HRQOL measures than did participants with osteoarthritis or rheumatoid arthritis. CONCLUSION: The BRFSS HRQOL measure is a brief, easily administered, generic health indicator that shows differences among rheumatic disease diagnoses. PMID- 14558052 TI - Adherence and satisfaction of rheumatoid arthritis patients with a long-term intensive dynamic exercise program (RAPIT program). AB - OBJECTIVE: To evaluate adherence and satisfaction of patients with rheumatoid arthritis (RA) in a long-term intensive dynamic exercise program. METHODS: A total of 146 RA patients started an intensive (strength and endurance training for 75 minutes, twice a week, for 2 years) exercise program (Rheumatoid Arthritis Patients In Training) aimed at improving physical fitness. Program attendance and satisfaction were examined. Additional assessments at baseline were done to find possible predictors of attendance. RESULTS: Median (interquartile range) age and disease duration of the patients were 54 (45-61) and 5 (3-10) years, respectively. After 2 years, 118 (81%) patients still participated in an exercise class. The median attendance rate of all patients was 74%. Low attendance was weakly associated with high disease activity, low functional ability, and low quality of life at baseline but not with the severity of joint damage at baseline. At the end, 78% of all participants would (strongly) recommend the program to other RA patients. CONCLUSION: Adherence and satisfaction of RA patients with an intensive dynamic exercise program over a prolonged time can be high. Disease severity parameters do not strongly predict the compliance of participants in an intensive exercise program. PMID- 14558053 TI - Correlates of falls and fear of falling among adults with rheumatoid arthritis. AB - OBJECTIVE: To identify correlates of falling and fear of falling, and to examine how fear of falling affects activities among adults with rheumatoid arthritis (RA). METHODS: Adults (mean age 54.2 years; SD 9.1) with RA (n = 128) responded to questions related to falls, fear of falling, and activities modified. Other measures included the Profile of Mood States Short Form, the McGill Pain Questionnaire Short Form, walk time, grip strength, predicted maximum oxygen uptake, and joint count. RESULTS: Thirty-five percent of participants fell during the previous year. Subjects who fell had more comorbid conditions than subjects who did not fall. Almost 60% were fearful of falling. Compared with subjects who denied fear of falling, fearful subjects had longer walk times, more comorbid conditions, and more intense pain. Activities affected most by fear of falling involved heavy work and climbing. CONCLUSIONS: The number of comorbid conditions plays an important role in falling and fear of falling in adults with RA. Knowledge of this and other factors, such as pain intensity and functional status, can facilitate appropriate interventions. PMID- 14558054 TI - Physical capacity in women with primary Sjogren's syndrome: a controlled study. AB - OBJECTIVE: To examine physical capacity (aerobic capacity, joint mobility, muscle function, and standing balance) in women with primary Sjogren's syndrome (primary SS) and to examine the correlation of aerobic capacity with fatigue, functional disability, and mental aspects (anxiety and depression). METHODS: Fifty-one women who fulfilled the European Community criteria for primary SS and who had anti SSA/SSB antibodies or a positive lip biopsy were compared with 51 age-matched controls. Physical capacity, fatigue, functional disability, anxiety, and depression were investigated by means of questionnaires and clinical examinations. RESULTS: The women with primary SS had significantly decreased aerobic capacity (VO(2max) = 28.7 versus 32.4 ml/kg/minute; P = 0.013), shoulder mobility (58 versus 59 scale points; P = 0.003), grip strength (214 versus 259 N; P = 0.000), isokinetic strength of the knee flexors (51 versus 56 Nm; P = 0.049), endurance of the knee flexors (620 versus 712 J; P = 0.008), and standing balance (25 versus 28 seconds; P = 0.006) when compared with the reference group. For the primary SS patients, greater effort was needed to carry out the test of aerobic capacity, and they experienced more pain during the shoulder mobility test. Aerobic capacity correlated with the fatigue experienced (r = -0.33, P = 0.022) but not with functional disability or mental aspects. CONCLUSION: The results indicate that women with primary SS have decreased physical capacity, which may be related to the experience of fatigue. PMID- 14558055 TI - Systemic sclerosis: patients' perceptions of their condition. AB - OBJECTIVE: To examine patients' beliefs about systemic sclerosis (SSc) and to investigate the relationship between these beliefs, symptom report, and clinical and demographic variables. METHODS: A total of 49 patients (7 male, 42 female) with SSc underwent clinical examination and completed the Revised Illness Perception Questionnaire. This measure assesses beliefs about symptoms, chronicity or recurrence of the condition, consequences, personal and treatment control, illness coherence, perceived causes of the condition, and patients' emotional response to their condition. RESULTS: The symptoms patients most frequently associated with their SSc were stiff joints (79%), pain (75%), and fatigue (75%). The most commonly reported causes of SSc were stress (53%), altered immunity (49%), and chance or bad luck (46%). More than 96% of patients believed that their condition would be chronic and 78% believed that the condition had serious consequences on their lives. Patients with diffuse cutaneous SSc reported more significant consequences of the condition and less personal control of their SSc compared with patients with limited cutaneous disease. There were no significant differences in illness beliefs between patients with nonsevere and severe ischemia. Multiple regression analyses indicated that illness beliefs, in particular perceived consequences associated with the condition, accounted for a significant proportion of the variance in emotional response to the condition. CONCLUSIONS: The beliefs held and symptoms experienced by patients with SSc are not ruled by disease subtype, skin score, functional ability, or severity of digital ischemia. This suggests patients' beliefs and emotional response are associated with the meaning they ascribe to their condition rather than its severity. PMID- 14558056 TI - Chronic upper extremity pain and co-occurring symptoms in a general population. AB - OBJECTIVE: To estimate the prevalence of self reported chronic upper extremity pain associated with physical impairment in a general population, and its co occurrence with chronic upper extremity numbness or tingling and chronic pain at other locations. METHODS: A general health questionnaire was mailed to 3,000 persons (age 25-74 years) who were randomly selected from a general population register. RESULTS: The response rate was 83%. The prevalence of chronic upper extremity pain associated with physical impairment was 20.8% (95% confidence interval [95% CI] 19.2-22.5), and that of co-occurring numbness or tingling was 6.7% (95% CI 5.7-7.7). Among the responders with chronic upper extremity pain associated with physical impairment, 84% reported more than 1 painful area. CONCLUSION: Chronic upper extremity pain associated with physical impairment and co-occurring chronic upper extremity numbness or tingling were common in the general population. The presence of more than 1 location for pain in the upper extremity as well as in other parts of the body was frequent. PMID- 14558057 TI - Glucocorticoid therapy in giant cell arteritis: duration and adverse outcomes. AB - OBJECTIVE: To evaluate the course of glucocorticoid (GC) therapy and associated adverse events in a population-based cohort of patients with giant cell arteritis (GCA). METHODS: We identified 125 Olmsted County residents with GCA diagnosed between 1950 and 1991 and obtained followup information on the 120 patients who were diagnosed antemortem and agreed to participate in this study. Clinical variables, GC doses, and GC adverse events on each patient were recorded. The relationship between GC therapy and the development of adverse events was studied by the Cox and Anderson-Gill proportional hazards models. RESULTS: All patients were treated with GCs and responded rapidly (median initial dosage 60 mg prednisone/day). The dosage was later reduced according to the treating physicians' judgment. The median duration required to reach 7.5 mg/day was 6.5 months and the median duration required to reach 5 mg/day was 7.5 months. Relapses or recurrences occurred in 57 patients. For the 87 patients followed to discontinuation of GC therapy and permanent remission of GCA (median of 22 months), the total median dose of prednisone was 6.47 gm. Adverse events associated with GCs were recorded in 103 (86%) patients and 2 or more events occurred in 70 patients (58%). Age and higher cumulative dose of GCs were associated with the development of adverse GC side effects. CONCLUSION: GCs are therapeutically effective in GCA and the prednisone dosage was reduced to physiologic levels in three-fourths of the patients within 1 year. However, most patients developed serious adverse side effects related to GCs, indicating that less toxic therapeutic measures are needed. PMID- 14558058 TI - Microscopic polyangiitis and polyarteritis nodosa: how and when do they start? AB - OBJECTIVE: To describe initial clinical symptoms attributable to microscopic polyangiitis (MPA) or polyarteritis nodosa (PAN). METHODS: We retrospectively reviewed the medical files of 72 patients (mean followup 6.7 years) with biopsy proven MPA (n = 36) or PAN (n = 36). RESULTS: Initial manifestations were similar in both entities except for peripheral neuropathy (P = 0.02) and gastrointestinal tract involvement (P = 0.006), which were significantly more frequent in PAN, and general signs alone in MPA (8%; P = 0.02). The mean time to diagnosis was 9.8 +/- 19.4 months; 35% of the patients died and 26% relapsed; significantly more MPA than PAN patients relapsed (P = 0.03). Time to diagnosis >/=90 days was associated with a trend toward more patients relapsing (P = 0.12), but not with an increased risk of mortality. CONCLUSION: Initial symptoms of MPA and PAN are usually nonspecific and last for several months before the diagnosis is made. A longer time to diagnosis is associated with a tendency to a higher relapse rate. PMID- 14558059 TI - A view of our future: the case for redesigning rheumatology practice. PMID- 14558060 TI - Translational research: getting the word and the meaning right. PMID- 14558061 TI - Clinical improvement in a patient with severe rheumatoid arthritis and chronic hepatitis B after prosorba column immunoadsorption: a one-year followup. PMID- 14558062 TI - Critical review of arthritis self-management strategy use. PMID- 14558063 TI - Update in osteoporosis. PMID- 14558064 TI - New standards for uric acid excretion and evidence for an inducible transporter. PMID- 14558066 TI - Neovascularization around the optic nerve in giant cell arteritis. PMID- 14558067 TI - Hodgkin's lymphoma presenting as Tietze's syndrome. PMID- 14558068 TI - Education for patients with rheumatoid arthritis: is it worth it? PMID- 14558071 TI - Can we accurately predict the development of rheumatoid arthritis in the preclinical phase? PMID- 14558072 TI - Guidelines for monitoring of methotrexate therapy: "evidence-based medicine" outside of clinical trials. PMID- 14558073 TI - On the track of neuropsychiatric lupus. PMID- 14558074 TI - The multiple facets of premature aging in rheumatoid arthritis. PMID- 14558078 TI - Antibodies against cyclic citrullinated peptide and IgA rheumatoid factor predict the development of rheumatoid arthritis. AB - OBJECTIVE: To evaluate the prevalence and predictive value of anti-cyclic citrullinated peptide (anti-CCP) antibodies in individuals who subsequently developed rheumatoid arthritis (RA) and to determine the relationship to rheumatoid factor (RF) of any isotype. METHODS: A case-control study was nested within the Northern Sweden Health and Disease Study and the Maternity cohorts of Northern Sweden. Patients with RA were identified among blood donors whose samples had been taken years before the onset of symptoms. Control subjects matched for age, sex, date of sampling, and residential area were selected randomly from the same cohorts. Anti-CCP antibody and RFs were determined using enzyme immunoassays. RESULTS: Eighty-three individuals with RA were identified as having donated blood before presenting with any symptoms of joint disease (median 2.5 years [interquartile range 1.1-4.7] before RA). In samples obtained before the onset of RA, the prevalence of autoantibodies was 33.7% for anti-CCP, 16.9% for IgG-RF, 19.3% for IgM-RF, and 33.7% for IgA-RF (all highly significant compared with controls). The sensitivities for detecting these autoantibodies >1.5 years and /=1 month with a stable, maximally tolerated dosage of oral MTX (/=30% maximum increase in the Cr level from baseline during the study, with the Cr level in 3 patients (3.8%) exceeding the range considered normal for their age and sex. The maximum Cr level during the study was 1.8 mg/dl. The ACR20 clinical response rate at the end of treatment was 52.5% (95% confidence interval 41.6 63.4%). CONCLUSION: In patients whose active RA persists despite treatment with MTX, tacrolimus in combination with MTX is safe and well-tolerated and provides clinical benefit. PMID- 14558081 TI - Monitoring by rheumatologists for methotrexate-, etanercept-, infliximab-, and anakinra-associated adverse events. AB - OBJECTIVE: To determine what monitoring protocols rheumatologists use to identify adverse events in rheumatoid arthritis (RA) patients treated with methotrexate (methotrexate), etanercept (etanercept), infliximab (infliximab), and anakinra (anakinra), how often rheumatologists encounter treatment-altering adverse events in their RA patients receiving these treatments, and how they feel about and comply with the current monitoring guidelines. METHODS: Three hundred ten physician members of the American College of Rheumatology (ACR) were notified by e-mail of a survey that was posted on our rheumatology Web site. Questions were closed-ended and multiple choice in format. RESULTS: One hundred twenty-three responses were received (40%). Most rheumatologists reported that they utilize the ACR recommended screening tests at baseline before methotrexate treatment is initiated. Seventy-nine percent reported that treatment-altering abnormalities had occurred in <5% of their methotrexate-treated RA patients, and 88% reported that such abnormalities had occurred in <10% of such patients, in the previous 3 years. Forty-one percent believed liver function monitoring guidelines need to be changed; 59% said they would agree with new guidelines that would include a recommendation for liver function monitoring every 3-4 months. Most rheumatologists were not aware of any guidelines for monitoring by blood tests in RA patients treated with biologic agents, yet the majority reported that they order blood tests before patients start these therapies and on followup. CONCLUSION: Our survey indicates that treatment-altering liver function abnormalities in methotrexate-treated RA patients are rare, and more than half of rheumatologists agree that a less stringent monitoring regimen should be considered. Rheumatologists and pharmaceutical companies might work together to develop guidelines for monitoring of patients treated with biologic agents. PMID- 14558082 TI - Polymorphisms in the interferon-gamma/interleukin-26 gene region contribute to sex bias in susceptibility to rheumatoid arthritis. AB - OBJECTIVE: To determine whether polymorphisms in the interferon-gamma (IFNgamma)/interleukin-26 (IL-26; formerly, AK155) gene cluster contribute to sex based differential susceptibility to rheumatoid arthritis (RA). METHODS: Four microsatellite markers, located in a 118-kb interval that contains both the IFNgamma and IL-26 genes on chromosome 12q15, were typed in 251 patients with RA and 198 unrelated healthy controls (all of whom lived in Northern Ireland) by means of polymerase chain reaction-based fragment analysis. RESULTS: Marker D12S2510, which is located 3 kb 3' from the IL-26 gene, was significantly associated with RA in women (corrected P [P(corr)] = 0.008, 2 degrees of freedom [2 df]) but not in men (P = 0.99, 2 df). A 3-marker haplotype, IFNGCA*13;D12S2510*8;D12S2511*9, was inferred that showed significant underrepresentation in women with RA (odds ratio 0.50, 95% confidence interval 0.32-0.78; P = 0.002, P(corr) = 0.03) but not in men with RA. CONCLUSION: Our results demonstrate that common polymorphisms in the IFNgamma/IL-26 gene region may contribute to sex bias in susceptibility to RA, by distorting the propensity of female carriers versus male carriers to contract this disease. These results conform to our recent observations of a role for this gene cluster in sex-based differential susceptibility to another Th1-type inflammatory disease, multiple sclerosis. PMID- 14558083 TI - Differential expression of HLA class II genes associated with disease susceptibility and progression in rheumatoid arthritis. AB - OBJECTIVE: Rheumatoid arthritis (RA)-associated HLA class II genes are assumed to promote susceptibility to and/or progression of the disease. Among the various modes of action proposed so far is the effect of the differential expression of HLA class II genes in different types of antigen-presenting cells on the Th1/Th2 balance. The aim of this study was to investigate the differential expression of genes encoded within the RA-associated HLA-DR4 superhaplotype and within the neutral DR7 and DR9 superhaplotypes. METHODS: The promoters encoded within these 3 haplotypes were first analyzed for sequence polymorphisms. To test for functional consequences, we assumed that the binding of nuclear factors to the promoter elements was correlated with the transcription activity, and we used surface plasmon resonance technology. To that end, oligonucleotides representing the polymorphic regulatory sequences and nuclear extracts from a monocyte cell line and a B cell line were used. RESULTS: While the promoters of the highly polymorphic HLA-DRB1*04, *07, and *09 alleles showed comparable binding of nuclear factors, differential binding was observed for the 2 promoters that drive the relatively nonpolymorphic DRB4 alleles in linkage disequilibrium with DRB1. Interestingly, analysis of RA patients positive for DR4, DR7, and DR9 revealed the segregation of radiographic progression with the stronger of the 2 DRB4 promoters, independent of the DRB1 allele. Moreover, DRB1*04 alleles in RA patients showed a reduced association with the DRB4 splice variant, completely preventing DRB4 expression. CONCLUSION: Our findings represent the first evidence of a correlation between the differential expression of HLA class II genes and both the susceptibility and the progression of RA. PMID- 14558084 TI - Galectin 3 and its binding protein in rheumatoid arthritis. AB - OBJECTIVE: To characterize the expression pattern and role of galectin 3 and galectin 3 binding protein (G3BP) in rheumatoid arthritis (RA), in comparison with galectin 1, and to explore whether soluble galectin 3 and G3BP, investigated in serum, synovial fluid, or cell culture supernatant, are associated with disease. METHODS: Synovial tissues from patients with RA or osteoarthritis (OA), as well as from healthy controls, were analyzed for galectins 1 and 3 and G3BP by in situ hybridization and immunohistochemistry. Levels of galectin 3 and G3BP in serum and synovial fluid from patients with RA and OA and controls, as well as in cell culture supernatants, were determined by enzyme-linked immunosorbent assay (ELISA). In vitro, the intracellular expression of galectin 3 in RA and OA synovial fibroblasts after modulation with tumor necrosis factor alpha (TNFalpha), interleukin-1beta (IL-1beta), and anti-CD40 monoclonal antibodies was measured by flow cytometry. RESULTS: In RA, galectin 3 messenger RNA and protein stained throughout the synovial membrane, whereas G3BP was particularly expressed at sites of bone destruction. In contrast, the expression of galectin 1 was not uniform in different RA specimens, and was never found at sites of invasion. In OA and normal synovial tissues, only a small number of cells were positive for galectins and/or G3BP. Galectin 3 was elevated in RA sera and synovial fluids, whereas G3BP was increased in RA synovial fluids only. In RA, serum galectin 3 correlated with C-reactive protein levels, whereas G3BP was associated with joint destruction and/or synovial cell activation as measured by the levels of cartilage oligomeric matrix protein. In vitro, RA synovial fibroblasts showed an increased release of galectin 3 into culture medium, as measured by ELISA, but decreased secretion of G3BP. In RA synovial fibroblasts with low basal expression of galectin 3, TNFalpha increased its intracellular level in a dose-dependent manner. In contrast, IL-1beta or anti-CD40 monoclonal antibodies showed no effect. CONCLUSION: Our data indicate that galectin 3 and G3BP are not only involved in inflammation, but also contribute to the activation of synovial fibroblasts. The intracellular accumulation of galectin 3 can be enhanced by TNFalpha. Thus, galectin 3 and G3BP represent novel markers of disease activity in RA. PMID- 14558085 TI - Synovial fluid neutrophils transcribe and express class II major histocompatibility complex molecules in rheumatoid arthritis. AB - OBJECTIVE: To investigate a potential interaction between neutrophils and T cells in rheumatoid arthritis (RA), by defining the optimal conditions for induction of class II major histocompatibility complex (MHC) expression on peripheral blood neutrophils in vitro and investigating the capacity for neutrophils to express class II MHC molecules in RA. METHODS: Surface expression of class II MHC and costimulatory molecules by peripheral blood and synovial fluid (SF) neutrophils obtained from healthy controls and patients with RA was measured by flow cytometry and fluorescence microscopy. Intracellular class II MHC protein and messenger RNA (mRNA) were detected by Western blotting and Northern blotting, respectively. RESULTS: Freshly isolated peripheral blood neutrophils from controls did not express surface class II MHC; expression was induced by culture with appropriate cytokines. Freshly isolated peripheral blood neutrophils from patients with RA expressed mRNA, but there was no surface expression of class II MHC. Freshly isolated SF neutrophils from patients with RA contained high levels of class II MHC mRNA, did not express surface class II MHC, but did have large intracellular amounts of this protein as detected by Western blotting. After culture for 20 hours in vitro, SF neutrophils from RA patients expressed large amounts of surface class II MHC but very low levels of costimulatory molecules CD80 and CD86. Fluorescence microscopy localized surface class II MHC to discrete areas on the neutrophil. Class II MHC-expressing neutrophils stimulated T cell proliferation. CONCLUSION: Peripheral blood neutrophils from patients with RA but not healthy controls express class II MHC mRNA. SF neutrophils in RA synthesize and express large amounts of class II MHC but not costimulatory molecules. This might underlie a novel interaction with T cells that is important in terms of disease pathology. PMID- 14558086 TI - Bone morphogenetic proteins 2 and 6, expressed in arthritic synovium, are regulated by proinflammatory cytokines and differentially modulate fibroblast like synoviocyte apoptosis. AB - OBJECTIVE: To examine the expression, regulation, and potential roles of bone morphogenetic proteins (BMPs) in arthritic synovium. METHODS: Expression of BMPs in arthritic synovium from patients with rheumatoid arthritis (RA) or spondylarthropathy (SpA) and in noninflamed synovium from patients undergoing diagnostic or therapeutic arthroscopies was studied by reverse transcription polymerase chain reaction (RT-PCR), Western blot, immunohistochemistry, and 2 color immunofluorescence. In vitro regulation of gene expression in fibroblast like synoviocytes (FLS) was determined by real-time quantitative RT-PCR and immunohistochemistry. We used (3)H-thymidine incorporation after serum deprivation-induced growth arrest to examine effects on FLS proliferation. FLS apoptosis was evaluated by flow cytometry cell cycle analysis. Apoptotic cells in synovial tissue were detected by TUNEL staining. RESULTS: Transcripts of different BMPs, most strikingly BMP-2 and BMP-6, were detected in synovial tissues. By Western blot, BMP-2 and BMP-6 precursor protein was found in RA and SpA synovial tissue extracts, but not in extracts of noninflamed synovial tissue. By immunohistochemistry, BMP-2 and BMP-6 were detected in the hyperplastic lining and the sublining layer of synovium from RA and SpA patients, both in CD90+ cells (FLS) and in some CD68+ cells (macrophages). Proinflammatory cytokines, such as interleukin-1beta and tumor necrosis factor alpha, but not interferon-gamma, enhanced the expression of BMP-2 and BMP-6 transcripts in FLS in vitro. Neither BMP-2 nor BMP-6 affected FLS proliferation. BMP-2 promoted FLS apoptosis, whereas BMP-6 protected against nitric oxide-induced FLS apoptosis. BMP-2-positive apoptotic cells were found in arthritic synovium. CONCLUSION: BMP-2 and BMP-6 are expressed in arthritic synovium and are strongly up-regulated by proinflammatory cytokines. Although BMP signaling has been proposed to be involved in cartilage and bone repair in arthritis, this pathway may be equally important in modulating FLS cell populations in inflamed synovium. PMID- 14558087 TI - Prostaglandin E2 is an enhancer of interleukin-1beta-induced expression of membrane-associated prostaglandin E synthase in rheumatoid synovial fibroblasts. AB - OBJECTIVE: Membrane-associated prostaglandin E synthase (mPGES) is a recently identified terminal enzyme of the arachidonic acid cascade, which converts PGH(2) to PGE(2) in rheumatoid arthritis synovial fibroblasts (RASFs). This study was undertaken to investigate factors regulating the expression of mPGES. METHODS: RASFs were treated with interleukin-1beta (IL-1beta), indomethacin, NS-398, rofecoxib, or meloxicam. The effects of PGE(2) and selective agonists for PGE(2) receptor subtypes (EP1, EP2, EP3, and EP4) were also studied. Expression of mPGES messenger RNA (mRNA) and protein was measured by Northern and Western blot analysis, respectively. EP receptor mRNA expression in RASFs was determined by reverse transcriptase-polymerase chain reaction. Production of PGE(2) and cAMP was measured by enzyme-linked immunosorbent assay. RESULTS: The enhanced expression of mPGES mRNA and protein in IL-1beta-stimulated RASFs was attenuated by the addition of indomethacin, NS-398, rofecoxib, or meloxicam. This reduction of expression was reversed by PGE(2). IL-1beta-induced PGES activity, measured by conversion of PGH(2) to PGE(2), was decreased by rofecoxib. EP2 and EP4 receptor mRNA was detected in RASFs. EP2 and EP4 agonists, as well as PGE(2), restored the inhibitory effect of rofecoxib on mPGES expression. The effect of PGE(2) was mimicked by forskolin, a direct activator of adenylate cyclase. Intracellular cAMP was increased by IL-1beta and was inhibited by rofecoxib. CONCLUSION: Enhancement of mPGES expression by PGE(2) via the EP2/EP4 receptors with an increase in cAMP may play an important role in articular inflammation in patients with RA. It also seems that cyclooxygenase 2 (COX-2) inhibitors decrease PGE(2) production not only by direct inhibition of COX-2, but also by reducing mPGES expression in activated RASFs. PMID- 14558088 TI - Feasibility of T and Z scores from magnetic resonance imaging data for quantification of cartilage loss in osteoarthritis. AB - OBJECTIVE: T scores (an indicator of the difference between patients and young healthy subjects) and Z scores (an indicator of the difference between patients and age-matched healthy subjects) are used in the diagnosis of osteoporosis and form the current basis for the definition of osteoporosis by the World Health Organization. We tested the feasibility of using T and Z scores derived from quantitative cartilage imaging with magnetic resonance imaging (MRI) for the diagnosis of osteoarthritis (OA). METHODS: High-resolution MR images of tibial cartilage were acquired from 126 young healthy adults (ages 20-35 years), 24 age matched elderly healthy adults (ages 50-75 years), 7 OA patients prior to tibial osteotomy, and 7 OA patients prior to knee arthroplasty. Cartilage volume, thickness, surface area, and original joint surface area (before onset of disease) were determined in the medial and lateral tibia. RESULTS: The cartilage volume of the medial tibia of osteotomy patients with varus malalignment displayed moderate T scores (-1.0), and more negative T scores (-3.8) were observed in knee arthroplasty patients with varus malalignment. Normalization of the cartilage volume to the original joint surface area substantially enhanced the scores in patients undergoing osteotomy (-2.3) and in patients undergoing knee arthroplasty (-5.5), and this was superior to the normalization ratios of cartilage volume to body height and cartilage volume to body weight, in terms of distinguishing the loss of articular cartilage. CONCLUSION: Quantitative analysis of OA by MRI is feasible using T and Z scores. However, cartilage volume should be normalized to the individual joint surface area in order to maximize the discriminatory power of this technique for the diagnosis of OA. PMID- 14558089 TI - Periarticular lesions detected on magnetic resonance imaging: prevalence in knees with and without symptoms. AB - OBJECTIVE: To evaluate, using magnetic resonance imaging (MRI), the prevalence of periarticular lesions in older persons with or without knee pain, and to assess the association of these lesions with knee pain. METHODS: Subjects ages 45 years and older, with or without knee pain, were recruited from Veterans Affairs medical centers and from the community. Weight-bearing posteroanterior, skyline, and lateral radiographs were obtained in all subjects. Subjects were divided into 3 groups: those with radiographic OA (ROA) and knee pain (n = 376), those with ROA and no knee pain (n = 51), and those with neither ROA nor knee pain (n = 24). A single knee (the more symptomatic one in subjects with knee pain) was imaged with a 1.5T scanner using T1- and T2-weighted and proton-density spin-echo imaging sequences. MRIs were read for the presence of periarticular lesions, which were categorized (according to their general location) as being either peripatellar (prepatellar, superficial infrapatellar, deep infrapatellar) or "other periarticular lesions" (semimembranosus-tibial collateral ligament bursitis, anserine bursitis, iliotibial band syndrome, tibiofibular cyst). RESULTS: Patients with knee pain had more severe radiographic disease than did subjects who were asymptomatic. Peripatellar lesions (prepatellar or superficial infrapatellar) were present in 12.1% of the patients with knee pain and ROA, in 20.5% of the patients with ROA and no knee pain, and in 0% of subjects with neither ROA nor knee pain (P = 0.116). However, other periarticular lesions were present in 14.9% of patients with both ROA and knee pain, in only 3.9% of patients with ROA but no knee pain, and in 0% of the group with no knee pain and no ROA (P = 0.004). CONCLUSION: Although peripatellar lesions are equally common among subjects with knee pain and those without knee pain, other periarticular lesions (including bursitis and iliotibial band syndrome) are significantly more common among subjects with knee pain and may contribute to pain in these individuals. PMID- 14558090 TI - Tumor necrosis factor alpha activation of the apoptotic cascade in murine articular chondrocytes is associated with the induction of metalloproteinases and specific pro-resorptive factors. AB - OBJECTIVE: Tumor necrosis factor alpha (TNFalpha) blockade provides substantive reduction of the symptoms of rheumatoid arthritis (RA). While the biologic actions of TNFalpha have been well characterized in immune and synovial cells, which are known to be major contributors to the progression of cartilage destruction in RA, the current studies were designed to assess the direct effects of TNFalpha on chondrocytes. METHODS: We examined the expression of several groupings of messenger RNA (mRNA) that define key biologic pathways that have previously been associated with either the general actions of TNFalpha or cartilage destruction, in murine articular chondrocytes isolated from wild-type mice and TNFalpha receptor-null (p55/p75(-/-)) mice. RESULTS: TNFalpha induced the expression of multiple mRNA that facilitate apoptosis and lead to apoptosis induced cell death. The induction of apoptosis was accompanied by the increased expression of several factors involved in the regulation of skeletal tissue proteolysis and resorption. Quantitative increases from 2-fold to >10-fold were seen for inducible nitric oxide synthase, matrix metalloproteinase 3, macrophage colony-stimulating factor, and osteoprotegerin mRNA expression. The dependence of the induction of these mRNA on TNFalpha was confirmed by comparison with the effects of TNFalpha on chondrocytes isolated from receptor-null mice. CONCLUSION: These findings demonstrate that TNFalpha alters the expression of a complex array of genes within murine chondrocytes that contribute to the destruction of joint surfaces, independent of its actions on synovial and immune cells. Further studies are needed to clarify the biologic actions of TNFalpha in human cartilage cells. PMID- 14558091 TI - Endothelin 1 promotes osteoarthritic cartilage degradation via matrix metalloprotease 1 and matrix metalloprotease 13 induction. AB - OBJECTIVE: Degradation of the collagenous extracellular matrix by metalloproteases (MMPs) plays an important role in the pathogenesis of osteoarthritis (OA). Recently, it was suggested that endothelin 1 (ET-1), a potent vasoconstrictor, may be involved in MMP regulation. This study investigated the role of ET-1 in OA cartilage degradation. METHODS: We explored ET-1 expression and synthesis in normal and OA cartilage and synovial membrane by reverse transcription-polymerase chain reaction and immunohistochemistry. MMP-1 and MMP-13 gene expression and protein synthesis were investigated using Northern blotting and enzyme-linked immunosorbent assays. Additionally, ET-1-induced collagenase activity, type II collagen metabolites, and tissue inhibitor of metalloproteases 1 (TIMP-1) protein were evaluated. RESULTS: We found expression and synthesis of ET-1, in situ, in both normal and OA cartilage and synovial membrane. We demonstrated that ET-1 induced gene expression and protein synthesis of both MMP-1 and MMP-13. These enzymes were produced in OA chondrocyte cultures, and the production increased in a dose-dependent manner in the presence of ET-1. In OA cartilage, ET-1 also induced type II collagen-derived neoepitopes concomitantly with an increase in collagenase activity and a decrease in TIMP-1 protein. CONCLUSION: Our results provide strong evidence of the catabolic role of ET-1 in OA cartilage via MMP-1 and MMP-13 up-regulation. As well, ET-1 increased the net MMP/TIMP balance and secondarily increased collagen degradation. Hence, ET-1 becomes an attractive factor to target in the conception of new therapeutic approaches for OA and other diseases in which MMP-13 and MMP-1 actions are crucial in tissue alteration. PMID- 14558092 TI - Hydrostatic pressure modulates proteoglycan metabolism in chondrocytes seeded in agarose. AB - OBJECTIVE: To investigate the effect of isolated hydrostatic pressure on proteoglycan metabolism in chondrocytes. METHODS: Bovine articular chondrocytes cultured in agarose gels were subjected to 5 MPa hydrostatic pressure for 4 hours in either a static or a pulsatile (1 Hz) mode, and changes in glycosaminoglycan (GAG) synthesis, hydrodynamic size, and aggregation properties of proteoglycans and aggrecan messenger RNA (mRNA) levels were determined. RESULTS: The application of 5 MPa static pressure caused a significant increase in GAG synthesis of 11% (P < 0.05). Column chromatography showed that this increase in GAG synthesis was associated with large proteoglycans. In addition, semiquantitative reverse transcriptase-polymerase chain reaction showed a 4-fold increase in levels of aggrecan mRNA (P < 0.01). CONCLUSION: Hydrostatic pressure in isolation, which does not cause cell deformation, can affect proteoglycan metabolism in chondrocytes cultured in agarose gels, indicating an important role of hydrostatic pressure in the regulation of extracellular matrix turnover in articular cartilage. PMID- 14558093 TI - Accelerated, aging-dependent development of osteoarthritis in alpha1 integrin deficient mice. AB - OBJECTIVE: Cell-matrix interactions regulate chondrocyte differentiation and survival. The alpha1beta1 integrin is a major collagen receptor that is expressed on chondrocytes. Mice with targeted inactivation of the integrin alpha1 gene (alpha1-KO mice) provide a model that can be used to address the role of cell matrix interactions in cartilage homeostasis and osteoarthritis (OA) pathogenesis. METHODS: Knee joints from alpha1-KO and wild-type (WT) BALB/c mice were harvested at ages 4-15 months. Knee joint sections were examined for inflammation, cartilage degradation, and loss of glycosaminoglycans (by Safranin O staining). Immunohistochemistry was performed to detect the distribution of alpha1 integrin, matrix metalloproteinases (MMPs), and chondrocyte apoptosis. RESULTS: In WT mice, the alpha1 integrin subunit was detected in hypertrophic chondrocytes in the growth plate and in a subpopulation of cells in the deep zone of articular cartilage. There was a marked increase in alpha1-positive chondrocytes in the superficial and upper mid-zones in OA-affected areas in joints from old WT mice. The alpha1-KO mice showed more severe cartilage degradation, glycosaminoglycan depletion, and synovial hyperplasia as compared with the WT mice. MMP-2 and MMP-3 expression was increased in the OA-affected areas. In cartilage from alpha1-KO mice, the cellularity was reduced and the frequency of apoptotic cells was increased. These results suggest that the alpha1 integrin subunit is involved in the early remodeling process in OA cartilage. CONCLUSION: Deficiency in the alpha1 integrin subunit is associated with an earlier deregulation of cartilage homeostasis and an accelerated, aging-dependent development of OA. PMID- 14558094 TI - Neuronal and astrocytic damage in systemic lupus erythematosus patients with central nervous system involvement. AB - OBJECTIVE: Symptoms originating from the central nervous system (CNS) frequently occur in patients with systemic lupus erythematosus (SLE). CNS involvement in lupus is associated with increased morbidity and mortality. Currently, reliable markers for activity in this condition are absent. The goal of this study was to determine the level of the light subunit of the neurofilament triplet protein (NFL) and that of glial fibrillary acidic protein (GFAP) in the cerebrospinal fluid of SLE patients with clinically verified CNS involvement and compare them with the levels in SLE patients without CNS involvement. METHODS: We assessed cerebrospinal fluid obtained from 99 patients with SLE and 99 age-matched controls for the presence of soluble molecules indicating neuronal destruction and astrogliosis-NFL and GFAP, respectively. Patients were evaluated clinically, with magnetic resonance imaging (MRI) of the brain, cerebrospinal fluid analyses, and neuropsychiatric tests. RESULTS: In the group of lupus patients with CNS involvement, intrathecal levels of NFL and GFAP were increased an average of 7 fold (P /=1.4 (logarithm of odds score <-2) (equivalent to a genetic contribution of >10% to the AS sibling recurrence risk ratio) within this area contributing to AS. CONCLUSION: These findings indicate that ANKH is not significantly involved in susceptibility to or clinical manifestations of AS. PMID- 14558097 TI - Experimental immunity to the G1 domain of the proteoglycan versican induces spondylitis and sacroiliitis, of a kind seen in human spondylarthropathies. AB - OBJECTIVE: Experimental immunity to the G1 domain of the cartilage proteoglycan (PG) aggrecan (AG1) leads to the development of spondylitis as well as polyarthritis in BALB/c mice. The PG versican contains a structurally similar G1 domain (VG1). This study was conducted to determine whether immunity to VG1 would elicit similar pathology in these mice. METHODS: Recombinant natively folded VG1 and AG1 were prepared. BALB/c mice received either a series of 5 injections of human VG1 or AG1, or no protein. Polyarthritis was determined clinically, and spondylitis and sacroiliitis histologically. Immunohistochemistry of rat tissues was used to study the localization of versican. Enzyme-linked immunosorbent assays were employed to study humoral immunity to the recombinant proteins as well as to overlapping synthetic peptides covering all these human G1 domains and mouse homologs. Affinity-purified antibodies to human AG1 and VG1 were isolated from sera of hyperimmunized mice. T lymphocyte proliferation assays were performed using recombinant human proteins. T cell lines reactive with specific immunodominant T cell epitopes in human AG1 and VG1 were isolated. Synthetic peptides encoding sequences in these human proteins and in corresponding mouse proteins were used in these analyses. Guanidinium chloride extracts of mouse spines were also used in Western blots to study antibody cross-reactivity. RESULTS: Immunity to recombinant VG1 did not result in clinical polyarthritis. There was, however, clear evidence that VG1, like AG1, could induce spondylitis in the lumbar spine and sacroiliitis. Accumulation of mononuclear cells was observed in spinal ligaments adjacent to the intervertebral disc, in the intervertebral disc, and in the sacroiliac joints, the same sites where versican is localized. In contrast to AG1-immunized mice, in which T cells reactive with human AG1 cross-reacted with mouse AG1, there was no evidence in VG1-immunized mice that T cell immunity to human VG1 was cross-reactive with a mouse synthetic peptide that contained the sequence corresponding to the single immunodominant T cell sequence recognized in human VG1. Antibodies to specific sequences in human VG1 did, however, cross-react with human AG1 and with corresponding peptide sequences in mouse versican and aggrecan and with mouse proteins containing VG1 and AG1, present in mouse spine extracts. Similarly, antibodies to human AG1 cross-reacted with human VG1 and with extracted mouse VG1 and AG1 and synthetic peptides containing mouse sequences that corresponded to the reactive human epitopes in AG1 and VG1. CONCLUSION: These observations suggest that humoral immunity to human VG1 is involved in the induction of experimental spondylitis and sacroiliitis in BALB/c mice. This humoral immunity is cross-reactive with mouse versican and aggrecan but is not associated with polyarthritis, probably because of the lack of cross-reactive T cell immunity and the absence of detectable versican in articular cartilage limbs. Induction of polyarthritis by bovine or human aggrecan requires the involvement of immunity mediated by T lymphocytes that are cross-reactive to a mouse aggrecan epitope. Together these observations suggest that humoral immunity to versican as well as immunity to aggrecan may be of importance in the development of the spinal pathology characteristic of spondylarthropathies. PMID- 14558098 TI - Subgrouping of fibromyalgia patients on the basis of pressure-pain thresholds and psychological factors. AB - OBJECTIVE: Although the American College of Rheumatology (ACR) criteria for fibromyalgia are used to identify individuals with both widespread pain and tenderness, individuals who meet these criteria are not a homogeneous group. Patients differ in their accompanying clinical symptoms, as well as in the relative contributions of biologic, psychological, and cognitive factors to their symptom expression. Therefore, it seems useful to identify subsets of fibromyalgia patients on the basis of which of these factors are present. Previous attempts at identifying subsets have been based solely on psychological and cognitive features. In this study, we attempt to identify patient subsets by incorporating these features as well as the degree of hyperalgesia/tenderness, which is a key neurobiologic feature of this illness. METHODS: Ninety-seven individuals meeting the ACR criteria for fibromyalgia finished the same battery of self-report and evoked-pain testing. Analyzed variables were obtained from several domains, consisting of 1) mood (evaluated by the Center for Epidemiologic Studies Depression Scale [for depression] and the State-Trait Personality Inventory [for symptoms of trait-related anxiety]), 2) cognition (by the catastrophizing and control of pain subscales of the Coping Strategies Questionnaire), and 3) hyperalgesia/tenderness (by dolorimetry and random pressure-pain applied at suprathreshold values). Cluster analytic procedures were used to distinguish subgroups of fibromyalgia patients based on these domains. RESULTS: Three clusters best fit the data. Multivariate analysis of variance (ANOVA) confirmed that each variable was differentiated by the cluster solution (Wilks' lambda [degrees of freedom 6,89] = 0.123, P < 0.0001), with univariate ANOVAs also indicating significant differences (all P < 0.05). One subgroup of patients (n = 50) was characterized by moderate mood ratings, moderate levels of catastrophizing and perceived control over pain, and low levels of tenderness. A second subgroup (n = 31) displayed significantly elevated values on the mood assessments, the highest values on the catastrophizing subscale, the lowest values for perceived control over pain, and high levels of tenderness. The third group (n = 16) had normal mood ratings, very low levels of catastrophizing, and the highest level of perceived control over pain, but these subjects showed extreme tenderness on evoked-pain testing. CONCLUSION: These data help support the clinical impression that there are distinct subgroups of patients with fibromyalgia. There appears to be a group of fibromyalgia patients who exhibit extreme tenderness but lack any associated psychological/cognitive factors, an intermediate group who display moderate tenderness and have normal mood, and a group in whom mood and cognitive factors may be significantly influencing the symptom report. PMID- 14558099 TI - Temperament and stress response in children with juvenile primary fibromyalgia syndrome. AB - OBJECTIVE: To examine temperament, stress response, child psychological adjustment, family environment, pain sensitivity, and stress response differences between children and adolescents with juvenile primary fibromyalgia syndrome (JPFMS), children with arthritis, and healthy controls. Parental psychological adjustment was also measured. METHODS: Subjects included 16 children with JPFMS, 16 children with arthritis, and 16 healthy controls. Participants completed the Dimensions of Temperament Survey-Revised (DOTS-R), State-Trait Anxiety Inventory, Children's Depression Inventory, Family Environment Scale (FES), Sensitivity Temperament Inventory for Pain (STIP), and Youth Self-Report. Responsiveness to an acute stressor was assessed by measuring salivary cortisol levels before and after venipuncture. Parents were asked to complete the parent versions of the DOTS-R, FES, STIP, Child Behavior Checklist, and Symptom Checklist-90-Revised. RESULTS: Children and adolescents with JPFMS demonstrated more temperamental instability, increased levels of depression and anxiety, less family cohesion, and higher pain sensitivity compared with the other 2 groups. Parents of children with JPFMS, in rating themselves, also reported higher levels of anxiety and depression, and lower overall psychological adjustment compared with parents of children in the other groups. CONCLUSION: These results suggest that a psychobiologic perspective may contribute to an increased understanding of JPFMS in children and adolescents, facilitating an approach to investigating the interaction of factors that appear to place a child at risk for development of a pain syndrome. Because temperamental instability, sensitivity to pain, vulnerability to stress, psychological adjustment, family context, and parental psychopathology are individual risk factors, the interaction of these factors may explain the breadth of symptoms associated with this pain syndrome, as well as its severity. PMID- 14558100 TI - Inhibition of monosodium urate monohydrate crystal-induced acute inflammation by retrovirally transfected prostaglandin D synthase. AB - OBJECTIVE: Hematopoietic prostaglandin D synthase (H-PGDS) is a key enzyme in the production of prostaglandin D and its J series metabolites. We evaluated the antiinflammatory effect of retrovirally transfected H-PGDS in order to investigate the role of H-PGDS in monosodium urate monohydrate (MSU) crystal induced acute inflammation. METHODS: Expression of endogenous PGDS in a murine air-pouch model of MSU crystal-induced acute inflammation was determined by real time polymerase chain reaction. H-PGDS complementary DNA (cDNA) was retrovirally transfected into C57BL/6J fibroblasts, and the cells were designated as C57-PGDS cells. Production of prostaglandins by C57-PGDS cells was measured by enzyme immunoassay. The effect of C57-PGDS cells on crystal-induced inflammation was investigated. RESULTS: Injection of the crystals caused a rapid decrease in H PGDS expression by infiltrating cells and by the soft tissues around the air pouches. In contrast, expression of interleukin-1beta (IL-1beta) and macrophage inflammatory protein 2 (MIP-2) as well as cellular infiltration were significantly increased during the early stage of inflammation. C57-PGDS cells, but not control cells, produced an increased amount of PGD(2) in vitro, but suppressed production of PGE(2). Injection of C57-PGDS cells into air pouches inhibited cellular infiltration and MIP-2 and IL-1beta expression. CONCLUSION: In this murine air-pouch model of MSU crystal-induced inflammation, retrovirally transfected H-PGDS cDNA could reduce cellular infiltration, at least partly by inhibiting MIP-2 and IL-1beta. These findings suggest that gene therapy with H PGDS may be useful for treating inflammatory diseases. PMID- 14558101 TI - Urinary type II collagen neoepitope as an outcome measure for relapsing polychondritis. AB - Herein we describe the case of a man who was diagnosed as having relapsing polychondritis (RP) when he was 18 years of age and was treated over the course of 2 years with numerous immunosuppressive agents, including tumor necrosis factor alpha (TNFalpha) inhibitors. His respiratory symptoms were refractory to treatment. Serum and urine samples were obtained periodically for measurement of erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) levels, anti-type II collagen (anti-CII) antibodies, and urinary type II collagen neoepitope (uTIINE) levels. The uTIINE assay is specific for collagenase cleavage products CII present in urine. ESRs and CRP levels varied widely but were rarely normal. Anti-CII antibody titers were high initially and decreased slowly and steadily for a year following the start of immunosuppressive medication, remaining low throughout the remainder of the patient's monitored disease course. The uTIINE levels were elevated prior to the initiation of TNFalpha inhibitors. Upon initiation of etanercept, they decreased abruptly to normal and stayed nearly normal. The uTIINE levels rose abruptly again upon discontinuation of TNFalpha inhibitor treatment. The dramatic decline in CII degradation, coincident with the administration of the TNFalpha inhibitors, suggested that this treatment dramatically reduced the chondritis. Serum levels of Th1 cytokines (interferon gamma, interleukin-12 [IL-12], and IL-2) paralleled changes in uTIINE levels, while those of Th2 cytokines (IL-4, IL-5, IL-6, and IL-10) showed little or no association with disease state or uTIINE levels. These results indicate that RP might be a Th1-mediated disease process. Moreover, the uTIINE assay appears to provide an objective measure of the severity of chondritis that could assist clinical decisions regarding adjustments of steroid and other immunosuppressive therapy. This outcome measure merits investigation in a broader spectrum of RP patients. PMID- 14558102 TI - Effectiveness of the soluble form of the interleukin-1 receptor accessory protein as an inhibitor of interleukin-1 in collagen-induced arthritis. AB - OBJECTIVE: To investigate whether the soluble form of interleukin-1 (IL-1) receptor accessory protein (sIL-1RAcP), whose physiologic function remains to be established, can serve as a specific inhibitor of IL-1 signaling in vitro, and to evaluate its applicability in collagen-induced arthritis (CIA). METHODS: Soluble IL-1RAcP was cloned from murine liver complementary DNA and expressed by the use of either an adenoviral vector (AdRGD) for sIL-1RAcP or a stable-transfected NIH3T3 fibroblast cell line. The ability of affinity-purified sIL-1RAcP to inhibit IL-1 signaling was tested on NF-kappaB luciferase reporter fibroblasts and quantified by luminometer. To investigate therapeutic efficacy, sIL-1RAcP was both locally (knee joint) and systemically overexpressed in collagen-immunized male DBA/1 mice. Severity of arthritis was monitored visually, and the pathologic process in the joint was examined histologically. Serum was obtained from mice to quantify IL-6 and anti-bovine type II collagen (BCII) antibody levels. RESULTS: Incubation of the NF-kappaB reporter fibroblast with purified sIL-1RAcP protein showed a marked reduction of IL-1-induced, but not tumor necrosis factor-induced, NF-kappaB activation. This showed a novel role for sIL-1RAcP as a specific inhibitor of IL-1 signaling. Local transplantation of sIL-1RAcP-producing NIH3T3 fibroblasts into the knee before onset of CIA had little or no effect on general disease severity in these mice. Histologic evaluation of the knee joints receiving sIL-1RAcP cell transplantation showed a marked reduction in both joint inflammation and bone and cartilage erosion. Local treatment with sIL-1RAcP had no profound effect on serum levels of IL-6 and anti-BCII antibodies, which is indicative of the ongoing presence of arthritis in distal joints. In contrast to local treatment, systemic treatment with the AdRGD for sIL-1RAcP markedly ameliorated CIA in all joints. CONCLUSION: In this study we demonstrated that sIL 1RAcP is a biologically active and innovative inhibitor of IL-1, and treatment of mice with sIL-1RAcP had a profound prophylactic effect on collagen-induced arthritis. PMID- 14558103 TI - Induction of arthritis in SCID mice by T cells specific for the "shared epitope" sequence in the G3 domain of human cartilage proteoglycan. AB - OBJECTIVE: To study the immunologic function and determine the fine epitope structure of a synthetic peptide p135H ((2373)TTYKRRLQKRSSRHP) of the G3 domain of human cartilage proteoglycan (aggrecan), which contains a highly homologous sequence motif of the shared epitope (QKRAA), the most common sequence motif in HLA-DR4 alleles, which predispose humans to the development of rheumatoid arthritis (RA). METHODS: Synthetic p135 peptides with altered sequences were used for (hyper)immunization of arthritis-susceptible BALB/c mice and then challenged with a single dose of cartilage proteoglycan. Human p135 (p135H) and mouse p135 (p135M) synthetic peptides of the G3 domain of aggrecan were used to prime lymphocytes, which were then used for adoptive transfer of arthritis into "presensitized" SCID mice, determining cross-reactivity among p135 peptides and their analogous sequences, and generating T cell hybridomas. T cell hybridomas were also used for arthritis transfer into SCID mice and for characterizing the fine epitope structure of T cell receptor (TCR) and major histo-compatibility complex (MHC) binding sites of the immunogenic/arthritogenic p135H sequence. RESULTS: While p135H peptide-(hyper)immunized mice became sensitized, they developed arthritis only after injection of a single dose of cartilage proteoglycan aggrecan. An altered peptide sequence (p135H-AA) carrying the shared epitope motif (QKRAA) was as effective as the natural peptide p135H sequence for inducing arthritis. Mouse p135M-specific lymphocytes induced arthritis with a lower incidence, but synthetic peptides to Escherichia coli heat-shock protein (DnaJ) or HLA-DR4 allele (both having the shared epitope sequence with different flanking regions) were also positive. Fine epitope sequence recognition of an arthritogenic T cell hybridoma derived from p135H-primed lymphocyte population was determined. Interestingly, in the most central position, a basic amino acid triplet of p135H peptide was found to be the MHC-binding motif, whereas the flanking amino acids bound to the TCR. CONCLUSION: Peptide p135H, corresponding to the peptide sequence in the G3 domain of human cartilage proteoglycan aggrecan, is immunogenic/arthritogenic in BALB/c mice. Peptide p135H includes a highly homologous motif of the shared epitope, a sequence that is overrepresented in bacterial heat-shock proteins, envelope protein of human JC polyomavirus, and numerous HLA-DR4 alleles. Since the G3 domain of cartilage proteoglycan aggrecan with the p135 sequence is "lost" during the normal metabolic turnover of cartilage proteoglycan or in pathologic conditions, an antigenoriented T cell migration into joints of presensitized (susceptible) individuals may contribute to the organ-specificity of RA. PMID- 14558104 TI - Reduction of atherosclerosis in low-density lipoprotein receptor-deficient mice by passive administration of antiphospholipid antibody. AB - OBJECTIVE: Patients with antiphospholipid syndrome (APS) are at a high risk of developing atherosclerotic complications. Conversely, individuals with primary atherosclerosis have an increased prevalence of antiphospholipid antibodies (aPL) and antibodies to oxidized low-density lipoproteins (ox-LDL). Several studies suggest that these two antibody populations may in fact overlap, although it is unclear how aPL contribute to pathogenesis. In this study, we characterized an IgG monoclonal aPL and assessed its ability to modulate atherosclerosis in low density lipoprotein receptor-deficient (LDLR(-/-)) mice. METHODS: The cardiolipin reactive monoclonal antibody FB1 was obtained from an (NZW x BXSB)F(1) mouse, a strain with APS features that make it prone to fatal myocardial infarctions. Using an enzyme-linked immunosorbent assay, we investigated the binding of this antibody to phospholipid and LDL antigens. We also passively administered FB1 to atherosclerosis-prone mice to determine its effect on atherogenesis. RESULTS: In contrast to earlier studies of aPL that were specific for oxidized forms of LDL, FB1 cross-reacted with both native LDL and ox-LDL. In vivo, passive administration of FB1 significantly reduced plaque formation in atherosclerosis prone LDLR(-/-) mice. CONCLUSION: These results indicate that some aPL may play a protective role in atherogenesis and suggest a novel approach to the prevention of atherosclerosis. PMID- 14558106 TI - Dependence on interferon-gamma for the spontaneous occurrence of arthritis in DBA/1 mice. AB - OBJECTIVE: Male DBA/1 mice are known to spontaneously develop arthritis in the hind legs. The present study was undertaken to investigate the role of endogenous interferon-gamma (IFNgamma) in the pathogenesis of this ankylosing enthesopathy. METHODS: The role of IFNgamma was studied by examining the development of arthritis in IFNgamma receptor-knockout (IFNgammaR-KO) DBA/1 mice as compared with wild-type mice, and by treatment of wild-type mice with monoclonal anti IFNgamma antibody. IFNgamma-disrupted and wild-type mice were mixed and housed in the same cage, and clinical symptoms of arthritis were assessed weekly for at least 9 weeks. Histologic examination was performed at the end of the experiment. RESULTS: In DBA/1 wild-type mice, 70% of the animals developed clinical symptoms of spontaneous arthritis, such as redness and swelling of the proximal interphalangeal joints, toe stiffness, and ankylosis. As evident from microscopic evaluation, the arthritis was mainly characterized by formation of new cartilage and bone, originating at the entheses and leading to ankylosis. The incidence and severity of arthritis, both clinically and histologically, were significantly reduced in IFNgammaR-KO mice. In wild-type mice, neutralizing anti-IFNgamma antibody inhibited the occurrence of the disease for the duration of treatment. CONCLUSION: The results suggest that endogenous IFNgamma plays an important role in the initial stages of spontaneous arthritis, and that the inflammatory components in its pathogenesis are more prominent than has been believed. In view of the similarity between this disease and spondylarthropathies in humans, the data suggest that endogenous IFNgamma may also play a disease-promoting role in the human condition and thus may serve as a target for therapy. PMID- 14558105 TI - The energy cost of adjuvant-induced arthritis in rats. AB - OBJECTIVE: To test the hypothesis that rats with adjuvant-induced arthritis (AIA) need more metabolic energy to maintain body weight than healthy control rats or rats with AIA treated with methotrexate (MTX). METHODS: Rat AIA was induced by Mycobacterium butyricum injection at the base of the tail. The MTX-treated group was injected with MTX (1.0 mg/kg/week) in phosphate buffered saline. Negative controls (i.e., disease-free) and the MTX-treated group were pair-fed with positive controls (i.e., untreated AIA rats) to ensure equal mean body weights. RESULTS: An additional 0.85 gm of food per day per rat was needed by the positive control group and 0.54 gm per day by the MTX-treated group to maintain a body weight comparable with that of the negative control group during days 5-15 post adjuvant injection. During days 15-34 post-adjuvant injection an additional 1.4 gm of food per day per rat was needed by the positive controls and 0.62 gm by the MTX-treated group. CONCLUSION: The results of this study indicate that adjuvant arthritis has a metabolic cost, which increases substantially as the disease becomes clinically apparent (days 15-34). MTX treatment does not completely eliminate the caloric cost of the disease. During days 5-15 post-adjuvant injection, an average of 6% of the total calories eaten by the positive controls was metabolized to support subclinical inflammation and other physiologic processes of this disease. During the active phase of the disease (i.e., clinical inflammation), this value increased to 18% of total calories. PMID- 14558107 TI - Lack of association between birth order and juvenile idiopathic arthritis. PMID- 14558108 TI - Ecotropic murine leukemia viruses and exogenous mouse mammary tumor viruses are not essential for pristane-induced lupus. PMID- 14558109 TI - Role of the H-2 haplotype in Fas-intact lupus-prone MRL mice: association with autoantibodies but not renal disease. PMID- 14558111 TI - Early changes in bone metabolism in rheumatoid arthritis patients treated with infliximab. PMID- 14558112 TI - Correlation between clinical response to treatment and radiographic progression: comment on the article by Strand and Sharp. PMID- 14558114 TI - Shrinking lung syndrome in systemic sclerosis. PMID- 14558115 TI - Clinical image: pseudoleukocytosis in cryoglobulinemia. PMID- 14558116 TI - A new dimension of 1H-NMR spectroscopy in assessment of liver graft dysfunction. AB - High-resolution 1H-NMR spectroscopy of serum and urine samples of an 11-year-old male living related orthotopic liver transplant recipient is reported. Serum glutamine increased to abnormal levels along with simultaneous abnormal excretion of urinary glutamine post-transplantation. High levels of glutamine in both blood and urine and concomitant reduced urea levels in urine were found to be evidence of impairment in urea cycle and compatible with persistently abnormal graft function. Thus glutamine levels in serum and urine, and urea in the urine as observed by 1H-NMR spectroscopy highlight their important roles in monitoring liver graft function; increased glutamine levels lead to brain damage, if untreated. PMID- 14558117 TI - Integration of [U-13C]glucose and 2H2O for quantification of hepatic glucose production and gluconeogenesis. AB - Glucose metabolism in five healthy subjects fasted for 16 h was measured with a combination of [U-13C]glucose and 2H2O tracers. Phenylbutyric acid was also provided to sample hepatic glutamine for the presence of 13C-isotopomers derived from the incorporation of [U-13C]glucose products into the hepatic Krebs cycle. Glucose production (GP) was quantified by 13C NMR analysis of the monoacetone derivative of plasma glucose following a primed infusion of [U-13C]glucose and provided reasonable estimates (1.90 +/- 0.19 mg/kg/min with a range of 1.60-2.15 mg/kg/min). The same derivative yielded measurements of plasma glucose 2H enrichment from 2H2O by 2H NMR from which the contribution of glycogenolytic and gluconeogenic fluxes to GP was obtained (0.87 +/- 0.14 and 1.03 +/- 0.10 mg/kg/min, respectively). Hepatic glutamine 13C-isotopomers representing multiply enriched oxaloacetate and [U-13C]acetyl-CoA were identified as multiplets in the 13C NMR signals of the glutamine moiety of urinary phenylacetylglutamine, demonstrating entry of the [U-13C]glucose tracer into both oxidative and anaplerotic pathways of the hepatic Krebs cycle. These isotopomers contributed 0.1-0.2% excess enrichment to carbons 2 and 3 and approximately 0.05% to carbon 4 of glutamine. PMID- 14558119 TI - Biochemical characterization of muscle tissue of limb girdle muscular dystrophy: an 1H and 13C NMR study. AB - The metabolic differences between the muscle biopsies of patients with limb girdle muscular dystrophy (LGMD) and normal controls were characterized using high-resolution 1H and 13C NMR spectroscopy. In all, 44 metabolites were unambiguously assigned in the perchloric acid extracts of skeletal muscle tissue, using 2D double quantum filtered (DQF COSY), total correlation (TOCSY), and 1H/13C heteronuclear multiple quantum coherence (HMQC) spectroscopy. The concentrations of glycolytic substrate, glucose (p=0.03), gluconeogenic amino acids, glutamine (p=0.02) and alanine (p=0.009) together with glycolytic product, lactate (p=0.04), were found to be significantly lowered in LGMD patients as compared with controls. The reduction in the concentration of glucose may be attributed to the decrease in the concentration of gluconeogenic amino acids in the degenerated muscle. Reduction in the rate of anaerobic glycolysis and lowered substrate concentration appear to be the possible reasons for the decrease in the concentration of lactate. A significant reduction in the concentration of choline in LGMD patients was also observed compared with controls. Lower concentration of choline may be the result of decreased rate of membrane turnover in LGMD patients. The data presented here provide an insight into the potentials of in vitro NMR spectroscopy in the study of muscle metabolism. PMID- 14558118 TI - Correlation between the occurrence of 1H-MRS lipid signal, necrosis and lipid droplets during C6 rat glioma development. AB - The aim of this study was to investigate the possible correlation between the 1H MRS mobile lipid signal, necrosis and lipid droplets in C6 rat glioma. First, the occurrence of necrosis and lipid droplets was determined during tumor development, by a histological analysis performed on 34 rats. Neither necrosis nor lipid droplets were observed before 18 days post-implantation. At later stages of development, both necrosis and lipid droplets were apparent, the lipid droplets being mainly located within the necrotic areas. Using a second group of eight rats, a temporal correlation was evidenced between mobile lipid signal detected by in vivo single-voxel one- (136 ms echo time) and two-dimensional J resolved 1H MR spectroscopy, and the presence of necrosis and lipid droplets on the histological sections obtained from the brains of the same rats. Finally, spatial distribution of the mobile lipid signal was analyzed by chemical-shift imaging performed on a third group of eight animals, at the end of the tumor growth. The spectroscopic image corresponding to the resonance of mobile lipids had its maximum intensity in the center of the tumor where necrotic regions were observed on the histological sections. These necrotic areas contained large amounts of lipid droplets. All these results suggest that mobile lipids detected in vivo by 1H MRS (136 ms echo time) in C6 rat brain glioma arise mainly from lipid droplets located in necrosis. PMID- 14558120 TI - Two-dimensional NMR spectroscopy of urinary glycosaminoglycans from patients with different mucopolysaccharidoses. AB - Patients with different types of mucopolysaccharidoses (MPS) lack specific lysosomal enzymes, which leads to tissue accumulation and urinary excretion of glycosaminoglycans (GAGs). Since little is known about the molecular composition of the excreted GAG fragments, we used two-dimensional [1H,13C]-correlation nuclear magnetic resonance (NMR) spectroscopy for a detailed analysis of the urinary GAGs of patients with MPS types I, II, IIIA, IVA and VI. The method revealed that the molecular structures of the excreted GAGs, i.e. heparan sulfate (HS), dermatan sulfate (DS), chondroitin sulfate (CS), and keratan sulfate (KS) are clearly distinct for the different MPS types. The chain terminal residues that are the normal substrates for the defective enzymes constitute characteristic sets of signals for each MPS type. The GAG chains show variations in carbohydrate composition and sulfation patterns that can be related to the different MPS types and clinical features. For example, two patients with MPS IIIA (M. Sanfilippo) with signs of CNS degeneration but only mild somatic features excrete a highly sulfated variant of HS, resembling HS in porcine brain, whereas a patient with MPS I (M. Scheie) and two patients with MPS II (M. Hunter), who present primarily with coarse facial features, joint contractures and skeletal deformities excrete a different type of HS with lower sulfation. In another case study, a patient with MPS IVA (M. Morquio), who presented mainly with skeletal dysplasia, excreted not only excessive amounts of KS but also a highly sulfated CS variant, resembling CS in articular cartilage. The high resolution NMR analysis of urinary GAGs presented here for the first time provides a solid basis for future studies with a larger number of patients to further explore pathogenesis and course of the MPS diseases. PMID- 14558121 TI - Identification of plant sterols in hexaploid and tetraploid wheats using gas chromatography with mass spectrometry. AB - Free sterols from hexaploid and tetraploid free-threshing wheats (Triticum aestivum L. and T. durum Desf.) and from their respective hulled wheats (T. spelta L. and T. dicoccon Schrank) were analysed by gas chromatography with mass spectrometry. The qualitative analysis of sterols showed a similar pattern either between hexaploid (T. aestivum, T. spelta) and tetraploid (T. durum, T. dicoccon) wheats or between free-threshing (T. aestivum, T. durum) and hulled (T. spelta, T. dicoccon) wheats. However, quantitative differences were found between tetraploid and hexaploid wheats, in that free sterol amounts in tetraploid wheats were 40% higher than in hexaploid ones. The mass spectra of the sterols were classified into four groups, taking into account the structural features of rings A and B. Typical mass spectral fragmentations of the four classes, and additional evidence related to the side chain of each molecule, were investigated together with their chromatographic behaviour, allowing identification of all the detected sterols. PMID- 14558122 TI - Analytical method for the quantitative determination of urinary ethylenethiourea by liquid chromatography/electrospray ionization tandem mass spectrometry. AB - A direct, rapid and selective method for the quantitative determination of the ethylenethiourea (ETU) in human urine has been validated and is reported in the present study. It allows the accurate quantification of ETU in this complex matrix without the use of any internal standard as the sample cleanup is effective enough for the removal of interferences that could lead to ion suppression in the electrospray ionization (ESI) source. This simple and rapid purification system, based on the use of a Fluorosil phase of a BondElut column followed by a liquid-liquid extraction procedure, achieves mean extracted recoveries, assessed at three different concentrations (2.5, 10.0, and 25.0 microg/L), always more than 85%. High-performance liquid chromatography (HPLC) with positive ion tandem mass spectrometry, operating in selected multiple reaction monitoring (MRM) mode, is used to quantify ETU in human urine. The assay is linear over the range 0-50 microg/L, with a lower limit of quantification (LOQ) of 1.5 microg/L and a coefficient of variation (CV) of 8.9%. The lower limit of detection (LOD) is assessed at 0.5 microg/L. The overall precision and accuracy were determined on three different days. The values for within- and between-day precision are < or = 8.3 and 10.1%, respectively, and the accuracy is in the range 97-118%. The relative uncertainties for the LOQ and QC concentrations have been estimated to be 18 and 8%, respectively. The assay was applied to quantify ETU in human urine from growers that regularly handle ethylenebisdithiocarbamate pesticides in large crop plantations. The biological samples were collected at the start and end of the working day, and the ETU urine levels were found to vary between 1.9 and 8.2 microg/L. PMID- 14558123 TI - Electrospray ion-trap multistage mass spectrometry for characterisation of co monomer compositional distribution of bacterial poly(3-hydroxybutyrate-co-3 hydroxyhexanoate) at the molecular level. AB - We report an electrospray ionisation multistage mass spectrometry (ESI-MSn) method that utilises molecular mass information for determination of sequence distribution and chemical structure of mass-selected macromolecules of poly(3 hydroxybutyrate-co-3-hydroxyhexanoate) biopolyester, PHBH. On the basis of ESI MSn studies of PHBH oligomers obtained by partial alkaline depolymerisation of natural PHBH containing 13-14 mol% of hydroxyhexanoate (HH) units, the microstructure of this bacterial copolyester was assessed up to the level of 28 repeat units. The subtle structural details of the PHBH were evaluated based on sequencing of individual macromolecular ions thus showing the utility of this technique for the analysis of biological copolyester macromolecules. It was confirmed that both HH and hydroxybutyrate (HB) units of the PHBH copolymer are randomly distributed. PMID- 14558124 TI - Phosphate buffers in capillary electrophoresis/mass spectrometry using atmospheric pressure photoionization and electrospray ionization. AB - Capillary electrophoresis (CE) has been combined with atmospheric pressure photoionization (APPI) and electrospray ionization (ESI) for mass spectrometric (MS) detection. Separation conditions using potassium phosphate buffer and ammonium formate buffer have been compared for analysis of eleven pharmaceutical bases. The results showed improvements in separation efficiency and peak symmetry when phosphate buffer was used. The low flow in CE may enable utilization of these advances with MS detection. Compared with ESI, the APPI technique provided a cluster-free background. The enhanced signal-to-noise ratio in the total ion current (TIC) and the reduced spectral background indicated that the APPI process is less affected by non-volatile salts in the CE buffers. This results in a wider range of choice of CE buffers in CE/MS analysis when APPI is the ionization method. PMID- 14558125 TI - Characteristic fragmentation behavior of phosphoamino acid conjugates with 3' azido-3'-deoxythymidine by electrospray ionization tandem mass spectrometry. AB - The conjugates of phosphoamino acids with 3'-azido-3'-deoxythymidine were synthesized and their structures were determined by various spectral methods. In positive and negative ion electrospray mass spectrometry (ESI-MS), the fragmentation pathways were investigated in conjunction with tandem mass spectrometry (MS/MS). The results showed that there were very different characteristic fragment ions in the positive ion MS/MS spectra and the negative ion MS/MS spectra. PMID- 14558126 TI - Induced in-source fragmentation for the selective detection of organic bound iodine by liquid chromatography/electrospray mass spectrometry. AB - The detection of unknown organic compounds, e.g., transformation products of known trace pollutants in environmental samples, is a difficult task which can be simplified if the compounds of interest are characterized by a functional group or a heteroatom which can selectively be detected. For the detection of iodinated benzene derivatives, used as X-ray contrast media, we have studied whether such compounds can be selectively detected by negative ion (NI) electrospray ionization via iodide as a product ion of in-source fragmentation, generated by applying a high cone voltage. It was found that selective detection of iodinated benzene derivatives is possible by liquid chromatography/negative ion electrospray ionization (LC/NI-ESI) if the compounds are destroyed before entering the mass spectrometer. To our knowledge this is the first report of the use of NI in-source fragmentation MS for analyses traditionally performed with inductively coupled plasma (ICP)-MS. The novel application of NI in-source fragmentation MS is a tool for those who do not possess the facilities to perform ICP-MS. Furthermore, NI-ESI at varying cone voltages provides both structural information and elemental detection, whereas ICP-MS can only provide elemental detection. PMID- 14558127 TI - Effects of hydrolysis on the delta13C values of individual amino acids derived from polypeptides and proteins. AB - This study investigates the effects of hydrolysis on the delta13C values of individual amino acids (IAAs) derived from polypeptide standards, and modern and ancient bone collagen. All IAAs were derivatised to their trifluoroacetyl/isopropyl (TFA/IP) esters for delta13C determination using gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS). Firstly, authentic single poly amino acid standards (SPAAs; n = 5) were hydrolysed for 4, 10, 24 and 48 h. As expected, IAA yields increased as a function of hydrolysis time. Significantly, it was only after 24 h of hydrolysis that IAA delta13C values were statistically identical to bulk SPAA values for all five standards. The accuracy of IAA delta13C values was thus shown to be a function of yield; however, poly phenylalanine demonstrated accurate IAA delta13C values with yields of only 1.4 and 4.3%, after 24 and 48 h of hydrolysis time, respectively. Authentic mixed poly amino acid standards (MPAAs; n = 5) comprising two different amino acids were then hydrolysed for 24 h. Percentage recoveries ranged from 36 95%. Estimates of bulk MPAA delta13C values calculated from measured IAA delta13C values agreed within experimental error with measured bulk MPAA values for three out of the five standards. Finally, the experimental procedure was applied to modern rat (MBCs; n = 20) and ancient ovi-caprine and bovine (ABCs; n = 27) bone collagen samples where the delta13C values of 12 out of its 18 constituent amino acids were determined. Estimated bulk MBC and ABC delta13C values were calculated from constituent amino acid delta13C values using mass balance. With the exclusion of three ABC samples, calculated bulk bone collagen delta13C values (delta13C(BCcal)) were shown to correlate extremely well with measured bone collagen values (delta13C(BCmes)) for both modern and ancient samples, where R2 = 0.91 and 0.84, respectively. Significantly, the variation between calculated and measured bone collagen values (Delta13C(BCcal-BCmes)) exhibited similar ranges for both MBC (from -2.6 to +1.2 per thousand ) and ABC (from -2.7 to +2.2 per thousand ) samples, providing evidence for the preservation of intact collagen in the ancient samples. These results demonstrate that the experimental procedures employed in the acid hydrolytic cleavage of peptides or proteins to their constituent amino acids does not involve significant isotopic fractionation. PMID- 14558128 TI - Linear quadrupoles with added octopole fields. AB - Two methods of adding relatively small octopole fields to the main quadrupole field of quadrupoles and linear ion traps with cylindrical rods are investigated. The first, 'stretching' the quadrupole by moving two rods out from the axis, produces a combination of higher order fields with similar magnitudes in which the octopole field is not necessarily the greatest. The quadrupole field strength is changed significantly and a large potential appears on the axis. The second method uses rod pairs of different diameters. It adds octopole components of up to several percent while all other higher order fields remain small. An axis potential is also added, but it is only a few percent of the radio-frequency (RF) voltage and approximately equal to the strength of the octopole field. The axis potential can be removed by moving the larger rod pair out from the axis or applying unbalanced RF to the electrodes. PMID- 14558129 TI - Quantification of the ratio of positional isomer dilinoleoyl-oleoyl glycerols in vegetable oils. AB - The distribution of fatty acids in a triacylglycerol is of great importance from nutritional, biochemical, quality and technological points of view. The ratio of triacylglycerol positional isomers containing two linoleic acid (18:2) and one oleic acid (18:1) moieties--namely, 1(3),2-dilinoleoyl-3(1)-oleoyl glycerol (LLO) and 1,3-dilinoleoyl-2-oleoyl glycerol (LOL)--were quantified in grape seed, olive, pumpkin seed, soybean, sunflower and wheat germ oils by high-performance liquid chromatography/atmospheric pressure chemical ionization mass spectrometry (HPLC/APCI-MS) in selected ion monitoring (SIM) mode. Relative LOL contents (LOL/(LLO+LOL)) of the oils were calculated from the mass abundances of the [LL]+ and [LO]+ diacylglycerol fragment ions ([M+H-RCOOH]+) using a calibration curve. The calibration curve of the relative diacylglycerol mass abundances was measured in SIM mode. The relative LOL contents were found to be relatively consistent for each oil variety. The relative LOL content in grape seed, sunflower, pumpkin seed, soybean and wheat germ oils accounted for 44.2 +/- 2.6, 26.8 +/- 3.2, 16.7 +/- 4.6, 15.9 +/- 2.9 and 13.9 +/- 4.3%, respectively. Only olive oils contained practically 100% of the LLO isomer. These results indicate that the unsaturated fatty acids such as linoleic and oleic acids have 'non-random' distribution patterns in various oils. PMID- 14558130 TI - Quantitative analysis of small pharmaceutical drugs using a high repetition rate laser matrix-assisted laser/desorption ionization source. AB - In this work, a high repetition rate laser matrix-assisted laser desorption/ionization (MALDI) source is studied on a quadrupole-time-of-flight (QqTOF) and a triple quadrupole (QqQ) mass spectrometer for rapid quantification of small pharmaceutical drugs. The high repetition rate laser allows an up to 100 fold higher pulse frequency as compared with regular MALDI lasers, resulting in much larger sample throughput and number of accumulated spectra. This increases the reproducibility of signal intensities considerably, with average values being around 5% relative standard deviation after taking into account the area ratio of the analyte to an internal standard. Experiments were conducted in MS/MS mode to circumvent the large chemical background due to MALDI matrix ions in the low mass range. The dynamic range of calibration curves on the QqTOF mass spectrometer extended over at least two orders of magnitude, whereas on the QqQ it extended over at least three orders of magnitude. Detection limits ranged from 60-400 pg/microL on the QqTOF and from 6-70 pg/microL on the QqQ for a series of benzodiazepines. The benzodiazepine content of commercial pill formulations was quantified, and less than 5% error was obtained between the present method and the manufacturer's certified values. Furthermore, a high sample throughput was achieved with this method, so that a single MALDI spot could be quantitatively scanned in as little as 15 s, and an entire 96-well MALDI plate in 24 min. PMID- 14558131 TI - A method for reducing the time required to match protein sequences with tandem mass spectra. AB - An algorithm for reducing the time necessary to match a large set of peptide tandem mass spectra with a list of protein sequences is described. This algorithm breaks the process into multiple steps. A rapid survey step identifies all protein sequences that are reasonable candidates for a match with a set of tandem mass spectra. These candidates are then used as models, which are refined by detailed analysis of the set of tandem mass spectra for evidence of incomplete enzymatic hydrolysis, non-specific hydrolysis and chemical modifications of amino acid residues resulting from either post-translational modifications or sample handling. Compared with current one-step methods for matching proteins to mass spectra, this multiple-step method can decrease the time required for the calculation by several orders of magnitude. PMID- 14558132 TI - Comparative study of matrix-assisted laser desorption/ionization and gas chromatography for quantitative determination of cocoa butter and cocoa butter equivalent triacylglycerol composition. AB - The triacylglycerol (TAG) composition study of cocoa butter (CB) and cocoa butter equivalents (CBEs) has been performed by gas chromatography (GC) and matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOFMS). These two techniques provided comparable results. The advantage of the MALDI technique was the detection of each compound comprising the triacylglycerol classes (Cn). Moreover, comparison of the data obtained by these two techniques indicated that TAG relative percentages could be obtained quantitatively with the MALDI technique. These techniques have been applied for the composition determination of CB + CBE mixtures. Encouraging results showed that it is possible to quantify an admixture containing as little as 4% of CBE. PMID- 14558133 TI - Trypsin digestion of proteins on intact immobilized pH gradient strips for surface matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis. AB - Isolelectric focusing (IEF) of proteins on immobilized pH gradient (IPG) strips is an integral part of two-dimensional (2D) electrophoresis-based proteomics. Proteins can be effectively analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) on the intact strip itself, leading to the creation of a virtual 2D map giving pI and MW information, bypassing the second dimension SDS-PAGE. Further, trypsin digestion of proteins on the strip can significantly aid the identification of IPG separated proteins. However, the small size of the peptides leads to diffusion along and outside the gel matrix. In this study, we describe a simple spray-based procedure to perform 'on-strip' trypsin digestion of proteins embedded in IPG strips. Examination of intact myoglobin and its tryptic peptides shows that post digestion diffusion of tryptic peptides is significantly minimized using this approach. PMID- 14558134 TI - Phenol, chlorobenzene and chlorophenol isomers: resonant states and dissociative electron attachment. AB - This paper reports a study of resonant dissociative electron attachment (DEA) to the phenol, chlorobenzene, p-, m-, and o-chlorophenol molecules. On the basis of spectroscopic and thermochemical approaches the resonant states of the molecular negative ions (NIs) and the structures of some dissociative decay products are assigned. In the electron energy range up to 3 eV, DEA processes are determined by the two 2[pi*]-shape resonances resulting mainly in formation of [M-H]- and/or Cl- ions. At higher electron energies the energy correlation between peaks in the negative ion effective yield curves and bands of UV spectra allowed identification of the core-excited resonances. The peculiarities of Cl- ion formation and the vibrational fine structure on the effective yield curves of the [M-H]- ions are discussed. The mass spectrometric procedures for measurement of relative cross sections for NI formation are described. PMID- 14558135 TI - PEAKS: powerful software for peptide de novo sequencing by tandem mass spectrometry. AB - A number of different approaches have been described to identify proteins from tandem mass spectrometry (MS/MS) data. The most common approaches rely on the available databases to match experimental MS/MS data. These methods suffer from several drawbacks and cannot be used for the identification of proteins from unknown genomes. In this communication, we describe a new de novo sequencing software package, PEAKS, to extract amino acid sequence information without the use of databases. PEAKS uses a new model and a new algorithm to efficiently compute the best peptide sequences whose fragment ions can best interpret the peaks in the MS/MS spectrum. The output of the software gives amino acid sequences with confidence scores for the entire sequences, as well as an additional novel positional scoring scheme for portions of the sequences. The performance of PEAKS is compared with Lutefisk, a well-known de novo sequencing software, using quadrupole-time-of-flight (Q-TOF) data obtained for several tryptic peptides from standard proteins. PMID- 14558136 TI - Production and fragmentation of multiply charged ions in 'electron-free' matrix assisted laser desorption/ionization. AB - An unusually large fraction of multiply charged ions is observed in 'electron free' matrix-assisted laser desorption/ionization (MALDI). Here we investigate how the yield of multiply charged ions depends on experimental parameters in MALDI. It is found to increase if measures are taken to limit the number of electrons in the plume, for example, by using non-metallic MALDI targets or low laser pulse energies. The ionization energy of the matrix is another important parameter that affects the yield of multiply charged ions: matrices with high ionization energies lead to greater intensities of multiply charged ions. It is furthermore proposed that some of the fragment ions observed in MALDI are due to reactions of analyte with electrons in the plume. The possibility of electron capture dissociation of multiply charged ions produced by MALDI is shown. PMID- 14558137 TI - Automated multiple-layer spotting for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of synthetic polymers utilizing ink-jet printing technology. AB - Recently, a new multiple-layer matrix-assisted laser desorption/ionization time of-flight mass spectrometry (MALDI-TOFMS) sample spotting technique for poly(ethylene glycol), offering improved analysis possibilities, was described. In this contribution the application of ink-jet printing to automated, multiple layer MALDI-TOFMS sample preparation of synthetic polymers is presented, allowing accurate deposition of matrix, additive and analyte solutions. The new sample preparation technique was evaluated for poly(ethylene glycol) as well as poly(methyl methacrylate) standards, and optimized settings for both synthetic polymers have been obtained. PMID- 14558138 TI - Matrix-assisted laser desorption/ionization time-of-flight analysis of low concentration oligonucleotides and mini-sequencing products. AB - A new sample preparation procedure has been developed to improve the matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) analysis of low concentration oligonucleotides. In this method, a few microL of the oligonucleotide solutions are first dispensed and allow drying and shrinking to a small spot on an anchoring target. Thereafter, a small volume (0.1 microL) of the saturated 3-hydroxypicolinic acid (3-HPA) solution is added to the top of the shrunk oligonucleotide spot. It is found that the sample prepared by this procedure is more homogenous and substantially reduces the need to search for 'sweet' spots. Importantly, the increased shot-to-shot and sample-to-sample reproducibility makes it possible to perform high-throughput and quantitative analysis. We demonstrated that this procedure allowed the robust detection of oligonucleotides at a level of 0.01 microM and mini-sequencing products produced using only 50 fmol of the extension primer. PMID- 14558139 TI - Gas-phase ligation of Cr+ and Fe+ with 4,9-diazapyrene. AB - The ligation of Cr+ and Fe+ with 4,9-diazapyrene (DAP) within a Fourier transform mass spectrometer was studied and compared with previous results for the ligands pyrene (Py) and 9-azaphenanthrene (Ap). The results confirm that Fe+, which reacts more slowly but in the same way as Cr+ to bind two Py ligands, behaves quite differently with the nitrogen heterocycles DAP and Ap: it reacts rapidly and it binds three and four of them, respectively. PMID- 14558140 TI - Chromatographic and mass spectral studies of perfluorooctanesulfonate and three perfluorooctanesulfonamides. AB - The chromatographic and mass spectral characteristics of perfluorooctanesulfonate (PFOS) and three nitrogen-substituted perfluorooctanesulfonamides have been obtained. A methyl/phenyl mixed-phase fused-silica capillary column was used for gas chromatographic (GC) analyses, while a C18 reversed-phase microbore column was used for liquid chromatographic (LC) analyses. Mass (MS) and tandem mass (MS/MS) spectra were generated using electron ionization (EI), argon CE, methane positive and negative ion CI, and ES ionization modes. EI spectra of the amides showed ions characteristic of both the fluorinated hydrocarbon and the sulfonamide portion of the molecules. The fragmentation pathway was studied using hydrogen/deuterium exchange, and was thought to involve a cyclic intermediate ion. Formation of molecular ions by CE and protonated molecule ions by CI to obtain molecular weight information was only partially successful. Negative ion ES-MS spectra provided intense [M-H]- anions for the amides, and an [M-K]- anion for PFOS from which molecular weight information could be obtained, while ES MS/MS produced product ions that could be used to detect the presence of these compounds in biological or environmental samples. PMID- 14558141 TI - Interactions between aminoglycoside antibiotics and carbohydrates studied by 252Cf plasma desorption mass spectrometry. PMID- 14558142 TI - MIG1-dependent and MIG1-independent regulation of GAL gene expression in Saccharomyces cerevisiae: role of Imp2p. AB - Imp2p (Yil154c) is a transcriptional activator involved in glucose derepression of the maltose, galactose and raffinose utilization pathways and in resistance to thermal, oxidative or osmotic stress. We analysed the role of Imp2 in the regulation of GAL genes. Imp2 was shown to have a positive effect on glucose derepression of Leloir pathway genes and their activator gene GAL4. The effect of Imp2 on galactose metabolism was shown to be partially dependent on Mig1p. The Mig1-independent role depends on Nrg1p. However, disruption of both MIG1 and NRG1 only partially relieves the glucose repression of GAL genes in the Deltaimp2 mutant, indicating that Imp2 must also have other function(s). Moreover, the interaction between IMP2 and GAL6/BLH1, a recently isolated gene involved in the regulation of GAL genes that shares with Imp2 the ability to protect cells from the glycopeptide bleomycin, was also analysed. The results suggest a major role of Imp2 in a GAL6-independent pathway. PMID- 14558143 TI - The ICL1 gene of Pichia pastoris, transcriptional regulation and use of its promoter. AB - We cloned and characterized a gene encoding isocitrate lyase from the methylotrophic yeast Pichia pastoris. This gene was isolated from a P. pastoris genomic library using a homologous PCR hybridization probe, amplified with two sets of degenerate primers designed from conserved regions in yeast isocitrate lyases. The cloned gene was sequenced and consists of an open reading frame of 1563 bp encoding a protein of 551 amino acids. The molecular mass of the protein is calculated to be 60.6 kDa with high sequence similarity to isocitrate lyase from other organisms. There is a 64% identity between amino acid sequences of P. pastoris Icl and Saccharomyces cerevisiae Icl. Northern blot analyses showed that, as in S. cerevisiae, the steady-state ICL1 mRNA levels depend on the carbon source used for cell growth. Expression in P. pastoris of the dextranase gene (dexA) from Penicillium minioluteum under control of the ICL1 promoter proved that P(ICL1) is a good alternative for the expression of heterologous proteins in this methylotrophic yeast. The sequence presented here has been deposited in the EMBL data library under Accession No. AJ272040. PMID- 14558144 TI - One-step measurement of firefly luciferase activity in yeast. AB - Firefly luciferase is often used as a sensitive genetic reporter in various cell types. The pitfall in yeast, however, has been the need to break down the rigid cells in order to measure the enzyme activity. In this study we have removed the peroxisomal targeting codons from the Photinus pyralis luciferase gene (luc) and shown that in the yeast Saccharomyces cerevisiae this modified luciferase gives high levels of light emission that is easy to measure from intact living cells. Furthermore, cells with the modified luciferase grew essentially faster than those with the wild-type luciferase, indicating that peroxisomal targeting of a foreign enzyme puts some constraints to cellular viability. As a model system we used two different reporter constructs. In the first, expression of the luciferase gene is under control of CUP1-promoter, a well known yeast promoter that is inducible by copper ions. In the second, luciferase activity is dependent on activation of the human oestrogen receptor and its interaction with oestrogen responsive elements incorporated in a yeast promoter. The luciferase activity measurement could be done on a 96-well plate by simple addition of the substrate, D-luciferin, at a moderately acidic pH of 5.0. The ease of use of the non peroxisomal luciferase makes it an interesting alternative for reporter genes that are conventionally used in yeast, such as lacZ. PMID- 14558145 TI - Role of the non-respiratory pathways in the utilization of molecular oxygen by Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae is a facultative anaerobe devoid of mitochondrial alternative oxidase. In this yeast, the structure and biogenesis of the respiratory chain, on the one hand, and the functional interactions of oxidative phosphorylation with the cellular energetic metabolism, on the other, are well documented. However, to our knowledge, the molecular aspects and the physiological roles of the non-respiratory pathways that utilize molecular oxygen have not yet been reviewed. In this paper, we review the various non-respiratory pathways in a global context of utilization of molecular oxygen in S. cerevisiae. The roles of these pathways are examined as a function of environmental conditions, using either physiological, biochemical or molecular data. Special attention is paid to the characterization of the so-called 'cyanide-resistant respiration' that is induced by respiratory deficiency, catabolic repression and oxygen limitation during growth. Finally, several aspects of oxygen sensing are discussed. PMID- 14558146 TI - Expression in Escherichia coli of a recombinant adenosine kinase from Saccharomyces cerevisiae: purification, kinetics and substrate analyses. AB - The Saccharomyces cerevisiae ADO1 gene is known to encode a homologue of eukaryotic adenosine kinases. This gene was expressed in Escherichia coli as a recombinant protein fused to a polyhistidine tag by using the rhamnose-inducible bacterial promoter rhaB. The recombinant protein was purified to apparent homogeneity and its ability to phosphorylate different substrates was evaluated. Adenosine (Km 3 microM) is its primary substrate. In addition, it also phosphorylates, albeit less efficiently, 3'-deoxyadenosine (cordycepin; Km 1.84 mM) and 3'-amino-3'-deoxyadenosine (Km 0.26 mM). Other kinetic properties of the recombinant enzyme have also been determined. PMID- 14558148 TI - Patient-specific treatment with variable torque straight-wire. PMID- 14558149 TI - Biology and mechanics of facial asymmetries in children and youths. AB - There is a need for further knowledge about the normal and abnormal growth patterns of bones and teeth. Genetic make-up, as well as various types of diseases and injuries such as trauma, inflammation, radiation and chemicals may affect skeletal and dental growth sites, thereby causing faulty growth. The degree of the subsequent deformity depends not only on the type, intensity, extent and chronology of the noxious agent, but also on the site and its particular involvement in growth activity. The knowledge of what happens to the stomatognathic system when some of its parts are altered is very important. Dentists are challenged to visualize the entire system in all its relationships: they must visualize the dynamics of how the system works. The relationship of the maxilla to the mandible and the mechanics of the stomatognathic system must be considered as a whole when considering the occlusion. The aim of this paper is to focus attention on the biologic basis of facial asymmetries and the functional reasons for their stabilization. PMID- 14558150 TI - The hidden power (and charm) of the Witzig winner speech. PMID- 14558152 TI - [The sanctimonious nurse and her patients]. PMID- 14558151 TI - "Real" early orthodontic treatment. From birth to age 8. AB - This article will introduce new concepts for earlier orthodontic (teeth) and orthopedic (jaw-bone) diagnosis and treatment from birth to age eight. While early orthodontic treatment around age 8 is getting some attention, age 8 is still too late to orthopedically guide proper jaw and airway growth in some children. By age 8, the jaws (maxilla and mandible) grow 80-90% of their adult size. Research shows that small jaws create small airways and increase the likelihood of life-threatening disorders, for life. Since the upper and lower jaw bones form the gateway to the human airway, a new earlier orthodontic protocol and standard is warranted. Unique Functional Jaw Orthopedic concepts will help form a new early orthodontic protocol. These new concepts may very well help general dentists, pedodontists and orthodontists move dental care into a future world of medical dentistry which will include airway development, bed-wetting, ear disease, heart disease and longevity. PMID- 14558153 TI - [Efficiency and costs containment]. PMID- 14558154 TI - [Anxiety--and no one reacts]. PMID- 14558155 TI - [From migration to satisfaction]. PMID- 14558156 TI - [Kidnapping obscures leadership]. PMID- 14558157 TI - [House calls by telephone]. PMID- 14558158 TI - [Interview with Christian Simonin. "Nursing personnel and social competence"]. PMID- 14558159 TI - [Measuring the quality of nursing diagnosis]. PMID- 14558160 TI - [Fear--when you see us!]. PMID- 14558161 TI - [Me, an abusive nurse? Never!]. PMID- 14558162 TI - [Do not do unto others....]. PMID- 14558163 TI - [The advantage of "field" activities]. PMID- 14558164 TI - ["Misery is the cause of more misery"]. PMID- 14558165 TI - [Efficiency and economics]. PMID- 14558166 TI - [Resources are not used optimally! Public and private care must cooperate for more prompt care of the patients]. PMID- 14558167 TI - [Mortality after myocardial infarction has decreased in nearly all Swedish counties during the 1990's. Greatest improvement seen in those counties with the worst initial results]. AB - In an international perspective, Sweden has a very low case fatality after acute myocardial infarction (AMI). The aim of this study was to present trends and regional differences in case fatality within 28 days of the first AMI for males and females in Sweden after adjusting for co-morbidity. Adjustments in order to remove random effects on the rank order of county councils were made. The study was based on national data on more than 500,000 cases of AMI. Between 1987 and 1999, case fatality after AMI decreased from 47% to 37% among men and from 44% to 34% among women. The case fatality in the individual counties varied from 29% to 37% for men and from 31% to 40% for women. Further analysis is needed in order to explain these variations. PMID- 14558168 TI - [Shorter waiting lists and better occupational environment! Organizational restructuring of the department of obstetrics and gynecology well worth the effort--less stress, excellent cooperation with primary care and satisfied patients]. AB - A structured approach to reducing waiting times was pursued at the department of obstetrics and gynecology at University Hospital (Akademiska sjukhuset) in Uppsala, one of the largest such departments in the country. One pillar in this restructuring has been a county-funded training course, with its procedures for evaluating one's organization with an eye to reducing waiting times. The result of this effort has been to reduce waiting times, while access to gynecological care in Uppsala has improved. This organizational restructuring has also facilitated an excellent working alliance with primary care and improved the work environment for physicians and staff. PMID- 14558169 TI - [The COMET study compares effect of two different beta-blockers in chronic heart failure. Carvedilol better than metoprolol]. PMID- 14558170 TI - [Shortages within Swedish psychiatry when it comes to suicide-prone patients. A survey of the Lex Maria cases is a warning signal]. PMID- 14558171 TI - [An impossible leader of a clinic. You are hospital chief executive officer]. PMID- 14558172 TI - [Investigation concerning animal experiments can limit medical research]. PMID- 14558173 TI - [A composed discussion about health care is wanted!]. PMID- 14558174 TI - [Cardiopulmonary resuscitation, being on call and afraid to commit a mistake]. PMID- 14558175 TI - [Don't make a blueprint of the Norwegian list patient system]. PMID- 14558176 TI - [Herbal and "health" preparations--an important cause of abnormal laboratory test results]. PMID- 14558178 TI - Approaches to combat with confounding by indication in observational studies of intended drug effects. AB - There has been a resurgence of controversy about the usefulness of observational data to study the efficacy of drugs. Nearly every week a researcher makes some criticism of clinical trials or justifies observational research into intended effects, with other researchers offering a contradictory viewpoint. Literature reviews are not useful in this regard because the contradictory studies will not usually be carried out. Some methods are discussed which may have potential utility in the study of intended effects. There may be a marginal role for statistical techniques such as propensity scores and confounder scores. More promising techniques may include ecological analyses, restriction of subjects and blinded prospective review. Because it is currently unknown when the observational study of drug efficacy is possible, we should arguably always carry out a study of the determinants of prescribing first, and possibly consider using the various techniques that are outlined in this article. PMID- 14558177 TI - The risk of liver disorders in women prescribed cyproterone acetate in combination with ethinyloestradiol (Dianette): a nested case-control study using the GPRD. AB - PURPOSE: To explore the risk of liver disorders associated with cyproterone acetate combined with ethinyloestradiol (CPA/EE). CPA/EE is licensed in the UK for the treatment of women with acne and hirsutism and is a treatment option for polycystic ovary syndrome (PCOS). It acts as a contraceptive also. METHODS: Using the General Practice Research Database, we conducted a cohort analysis and case control study in women aged 15-39 with acne, hirsutism or PCOS to estimate the risk of liver disorders associated with CPA/EE. RESULTS: Compared with cases exposed to conventional combined oral contraceptives (COCs), the age-adjusted incidence rate ratio for liver disorders in women using CPA/EE was 1.7 (95% CI: 0.9, 3.4) and compared with no use it was 1.5 (95% CI: 0.8, 2.8). In the case control study, the adjusted odds ratio (OR) for liver disorders in women exposed to CPA/EE was 1.6 (95% CI: 0.7, 3.5) and 0.8 (95% CI: 0.5, 1.3) for exposure to conventional COCs, compared with no use. The risk of liver disorders in women prescribed CPA/EE was not significantly greater than that in women prescribed conventional COCs (OR: 2.1 [95% CI: 0.9, 4.8]). CONCLUSION: Our results do not indicate an increased risk for liver disorders associated with CPA/EE use in women with acne, hirsutism or PCOS after adjusting for potential confounding. This may be due to lack of statistical power. PMID- 14558179 TI - Practical pharmacovigilance analysis strategies. AB - PURPOSE: To compare two recently proposed Bayesian methods for quantitative pharmacovigilance with respect to assumptions and results, and to describe some practical strategies for their use. METHODS: The two methods were expressed in common terms to simplify identifying similarities and differences, some extensions to both methods were provided, and the empirical Bayes method was applied to accumulated experience on a new antihypertensive drug to elucidate the pattern of adverse-event reporting. Both methods use the logarithm of the proportional risk ratio as the basic metric for association. RESULTS: The two methods provide similar numerical results for frequently reported events, but not necessarily when few events are reported. Using a lower 5% quantile of the posterior distribution gives some assurance that potential signals are unlikely to be noise. The calculations indicated that most potential adverse event-drug associations that were well-recognized after 6 years of use could be identified within the first year, that most of the associations identified in the first year persisted over time. Other insights into the pattern of event reporting were also noted. CONCLUSION: Both methods can provide useful early signals of potential drug-event associations that subsequently can be the focus of detailed evaluation by skilled clinicians and epidemiologists. PMID- 14558180 TI - Regional variation in the use of medications by older Canadians--a persistent and incompletely understood phenomena. AB - BACKGROUND: We have previously reported on regional variability in medication consumption by older Canadians. In this study, we used longitudinal data to determine whether regional differences in commonly consumed medications persisted and to explore potential explanatory factors for observed differences. METHODS: We utilized data from the second phase of the Canadian Study of Health and Aging to assess the number, types, and variability of medications used between regions. Linear and logistic regressions (LRs) were used to predict the number of medications and the use of specific agents where significant regional variability was found to exist. RESULTS: There were significant regional differences in the number of medications consumed and in the prevalence of use of acetaminophen (p < 0.002), benzodiazepines (p < 0.020), nitrates (p = 0.040), and complementary and alternative medicines (CAMs; p < 0.020). The proportion of subjects using acetaminophen was highest in British Columbia (44.6%) and lowest in Quebec (27.3%). Benzodiazepine and nitrate consumption was highest in Quebec (35.9 and 19%, respectively) and lowest in the Praires (18.2%) and Atlantic Canada (6.6%). CAM use was highest in British Columbia (47.1%) and lowest in the Atlantic region (26.8%). Similar inter-regional differences had been found 5 years previously. There were no significant regional differences in the prevalence of hypertension, myocardial infarction, diabetes, arthritis/rheumatism, or depression. Region remained a significant explanatory variable for the number of medications and nitrate, benzodiazepine, and CAM use in our multivariate models. CONCLUSIONS: Regional differences in medication use persisted over the course of this longitudinal study. Much of the variability remains unexplained. The reasons for regional differences in consumption of drugs and their clinical significance should be addressed. PMID- 14558181 TI - Users of oral steroids are at a reduced risk of developing irritable bowel syndrome. AB - PURPOSE: To study whether irritable bowel syndrome (IBS) is associated with the use of oral steroids and whether there is a dose- or duration-response. METHODS: We followed up a cohort of 65,270 patients aged 20-74 years old enrolled in the General Practice Research Database in the UK with at least one prescription for steroids between 1994 and 1999. We performed a nested case-control analysis to estimate the adjusted relative risk (RR) associated with the use of steroids using unconditional logistic regression. Cases were 466 patients with a first episode of IBS during follow-up and controls were 5000 individuals randomly selected from the study cohort. RESULTS: Current users of oral steroids presented an RR of 0.6 (95% CI: 0.4-0.9) compared to non-users. Doses greater than 10 mg of prednisolone daily were associated with an RR of 0.4 (95% CI: 0.2-0.9). When we stratified by age, no reduced risk of IBS was apparent under the age of 40 years. The reduced risk of IBS was greater among females than males. CONCLUSIONS: Our results suggest that oral steroids can reduce the risk of a diagnosis of IBS. The apparent effect modification of age and sex deserves further research. PMID- 14558182 TI - Randomized controlled intervention in cardiovascular drug treatment in nursing homes. AB - OBJECTIVE: To study drug treatment of patients with cardiovascular diseases (heart failure, post-myocardial infarction, angina pectoris, hypertonia or cardiac valvular disease) in nursing homes and assess effect of medication advice. INTERVENTION: The patients were randomized to an intervention or control group. Medication reviews were made by one specialist in clinical pharmacology and one in cardiology. Symptoms related to heart failure or adverse reactions to cardiovascular drugs were recorded using a questionnaire. Quality of life and activities of daily living (ADL) were assessed and follow-ups performed after 2 weeks and 3 months. Outcome measures were changes of drug therapy and the global scores computed from symptoms scales. RESULTS: Eighty patients were randomized. They had a mean age of 87 years and their average number of drugs was 9.6. Changed drug therapy was suggested in 40 patients and the advice was followed by the responsible physicians in 19 patients. The physicians mostly followed advice for changed furosemide therapy but not for introducing an ACE-inhibitor, probably due to uncertain diagnosis and need for follow-up after initiation of such therapy. No significant changes from baseline to later follow-up were found in the mean total scores of any questionnaire. CONCLUSION: Intervention did not affect cardiovascular symptoms. Drug revisions should involve more than one class of drugs in order to be cost-effective. PMID- 14558183 TI - Pattern of prescriptions issued by nursing home-based physicians versus office based physicians for frail elderly patients in German nursing homes. AB - PURPOSE: The purpose of this study was to describe the physician prescription pattern for frail elderly patients in German nursing homes and to identify differences, if any, between that of physicians based in the nursing homes (A) and those with office-based practices (B). METHODS: Retrospective, longitudinal study of medication prescriptions on the basis of the database of a health insurance (Betriebskrankenkasse) in Berlin, Germany. We assessed the medication prescriptions for all 996 unselected insured individuals aged > or = 60 years who were insured throughout 1999 and lived in nursing homes. We analyzed selected patient characteristics and prescription data. Drugs were classified according to the WHO ATC code and were assigned a mean daily defined dose (DDD). RESULTS: 816 individuals were women and 180 were men. A total of 78% of women and 43% of men were aged 80 years or older. Two hundred sixty three patients were seen by nursing home-based physicians (A) and 733 by office-based physicians (B). The median of prescriptions per patient and year was 31 (A) and 37 (B) (p between groups < 0.001). Patients in Group A also received a lower number of DDDs in comparison to Group B (1109 vs 1250; p < 0.01). Costs were substantially higher in group B, and in both groups higher in the 60-69 years old in comparison to the more senior patients. The prescription pattern suggested a considerable frequency of inappropriate drug use in both groups in the following classes: psychopharmacological agents (neuroleptics, antidepressants, hypnotics), pain medication, digitalis glycosides, laxants and loop diuretics. On an average, office-based physicians (B) prescribed relatively more medications in all major classes with the exception of non-opiate analgesics, laxants and anxiolytics. CONCLUSIONS: These data are indicative of a considerable use of inappropriate medication for frail geriatric patients. Differences between the prescribing pattern among nursing home-based and office-based physicians were not substantial, however, the latter group prescribed relatively more drugs. PMID- 14558185 TI - Benefit-risk analysis: a proposal using quantitative methods. AB - This is the first part of a two-article series which will introduce the theory and practice of a proposed set of quantitative methods for benefit-risk analysis. Adjustments to number-needed-to-treat (NNT) analysis and a new method, minimum clinical efficacy (MCE) analysis are presented and critically discussed. The goal of these methods is to condense into a summary metric the benefit-risk profile of a product so that manufacturers, regulators, clinicians and patients can better understand and participate in risk management. A second article will present examples of these methods. PMID- 14558184 TI - Risk factors for adverse events in analgesic drug users: results from the PAIN study. AB - BACKGROUND: The relative influence of various risk factors for adverse events (AE) in analgesics users have never been precisely quantified. Advantage was taken of data generated in the paracetamol, aspirin and ibuprofen new tolerability (PAIN) study, a large randomized double-blinded trial of paracetamol, aspirin or ibuprofen for common pain in general practice to attempt this. OBJECTIVE: Identify and quantify factors associated with the occurrence of AE in users of analgesic drugs. METHOD: Multivariate logistic regression analysis of potential risk factors for all AE, clinically significant AE (SAE) and clinically significant gastro-intestinal AE (GI SAE). RESULTS: Of the 8677 patients included in the study, 8633 contributed data. The main risk factors for SAE were indication: compared to those treated for musculoskeletal pain, patients treated for menstrual pain had an odds ratio (95% Confidence Interval) of 0.4 (0.2-0.7), sore throat 0.6 (0.5-0.8), cold and flu 0.7 (0.6-0.8), headache 0.8 (0.7-1.0); concomitant use of medication contra-indicated in the drugs' labeling (OR: 2.2; 1.6-2.9); increasing number of other concomitant medications: 1: OR 1.5 (1.3-1.8); 2-3: OR 1.9 (1.6-2.3); more than 3: OR (2.7; 2.1-3.5); treatment with aspirin: OR 1.4; (1.2-1.6) but not ibuprofen: OR 0.9; (0.8-1.1) compared to paracetamol; history of previous GI disorder OR 1.4 (1.0-1.8); female gender: OR 1.3 (1.1-1.4). Age was not significantly associated with AE in the multivariate analysis. Risk factors for all AE and GI SAE were mostly the same as for significant AE, but there were fewer GI SAE with ibuprofen than with paracetamol (OR 0.8; 0.6-0.9). CONCLUSION: Apart from the analgesic used and its indication, the main risk factors identified for AE in users of first-line analgesics for common pain were the number and nature of concomitant medication. PMID- 14558186 TI - Pharmacoepidemiology and drug safety. PMID- 14558187 TI - [Progesterone+estrogen increased the risk of dementia in postmenopausal women. Surprising results of the WHIM study]. PMID- 14558188 TI - [Exercise is beneficial for patients with myositis. Both pharmaceuticals and physical activity should be included in the therapy of chronic rheumatic muscle inflammation]. PMID- 14558190 TI - [New European diagrams for assessment of cardiovascular risk]. PMID- 14558189 TI - [Undiagnosed depressions everywhere in the world. Somatic outpatient and inpatient care and primary health care in Uppland screened]. PMID- 14558191 TI - [A compilation of questionnaires on medical education. A specific part of education is always more valued because of its significance than because of its quality]. PMID- 14558192 TI - [Principles of family practice can be inspirations to improved care and occupational environment. Future primary health care benefits when focusing on the patient]. PMID- 14558193 TI - [Eric Hallstrom--the naivist who could enter into the sick children's world]. PMID- 14558194 TI - [Medical safety at our airports--time for an emergency team at the Arlanda airport]. PMID- 14558195 TI - [A "citizen-salary" and community service can activate persons on prolonged sick leave]. PMID- 14558196 TI - [A side-kick against the sick-listed]. PMID- 14558197 TI - [Internal medicine in future community health care--don't forget the elderly!]. PMID- 14558198 TI - [Albinism in Tanzania]. PMID- 14558199 TI - Paediatric dentistry in the new millennium: 3. Use of inhalation sedation in paediatric dentistry. AB - Provision of General Anaesthesia is now limited and restricted to the hospital setting. Sedation for paediatric patients is an essential tool in anxiety management and is used as an adjunct to behaviour management. Inhalation sedation with nitrous oxide/oxygen sedation to reach a plane of relative analgesia may be administered easily and safely to children in general dental practice and is a potential alternative to general anaesthesia. PMID- 14558200 TI - Occlusion: I. Terms, mandibular movement and the factors of occlusion. AB - This short series of three papers will review the relationship between mandibular movement and tooth morphology during function and dysfunction. ICP, RCP and long centric are discussed as is a description of mandibular excursions. Incisal guidance, condylar guidance, sagittal curve of Spee and lateral curve of Monson are inter-linked to illustrate how these factors of occlusion influence occlusal anatomy. PMID- 14558201 TI - Direct core materials. AB - There are many materials that can be used for direct-placement cores. Although the scientific evidence is incomplete, some materials are better suited to this task than others. This article provides an overview of direct-placement core materials and highlights what clinicians should consider when assessing a new product. PMID- 14558202 TI - An investigation of the effect on 27-gauge needle tips following a single local anaesthetic injection. AB - One hundred and forty one, 27-gauge local anaesthetic needles were collected, each following a single injection of local anaesthetic using four commonly performed local anaesthetic injection techniques. The needles were examined for needle tip damage under the scanning electron microscope. Ten unused needles were used as controls. Bone was contacted in 75 (59%) instances and, of these, 73 (97.3%) showed needle tip deformity. Of the four techniques used, bone contact was commonest with the inferior alveolar nerve block, occurring in 88% of instances. PMID- 14558204 TI - Organizing a practice-based research group. AB - There is an urgent need for more dental research to come from the 'real-life' setting of general practice in order to complement the information arising from laboratory testing and institution-based clinical trials. This paper outlines some of the problems and opportunities encountered in undertaking research in practice, and describes the experiences of one such practice-based research group. PMID- 14558203 TI - Nerve damage and third molar removal. AB - The surgical removal of lower third molars endangers both the lingual and inferior alveolar nerves. Patients sustaining an injury to either of these nerves must be managed correctly, and this requires a diagnosis of the injury type and regular monitoring of the recovery of sensation. Surgical intervention for a damaged inferior alveolar nerve is not usually indicated but may be undertaken: if the nerve is completely divided and the severed ends are misaligned; if a bony fragment has compressed the mandibular canal; or if the patient suffers from persistent neuropathic pain. In contrast, after injury to the lingual nerve, if sensory testing demonstrates no neural recovery within 3-4 months, exploration of the injury site and microsurgical repair of the damaged nerve is indicated. PMID- 14558205 TI - Tooth-supported stud-retained prostheses: three case reports. AB - The use of teeth as overdenture abutments is a common form of treatment. However, most roots are used only fur support. Simple stud precision attachments will also aid the retention of the prosthesis. This article presents three different cases where studs have been used to help retain removable prostheses. PMID- 14558206 TI - Preparing pictures for publication: guidelines for dental update. AB - To assist submission of articles to Dental Update in the correct format and thereby speed up acceptance of your article, Dental Update offers these guidelines for preparing and submitting illustrations. PMID- 14558207 TI - Physical signs for the general dental practitioner. PMID- 14558208 TI - [Calories more important than carbohydrates for reducing weight]. PMID- 14558209 TI - [Mass screening for prostate cancer will have to wait. The scientific basis insufficient, more efficient markers necessary]. PMID- 14558210 TI - [Too early to recommend universal screening for colorectal cancer]. AB - Randomised controlled trials have shown that screening may significantly reduce mortality in colorectal cancer. In the present evaluation it is shown that the reported reduction of 15 per cent, or higher, is given in relative figures. With absolute figures the reduction is only 0.15 per cent or lower. The number of individuals needed to screen (NNT) biannually for 10 years to prevent one death in colorectal cancer is 1,173 (confidence interval 741-2,807). Side-effects, costs and ethical issues are seldom, or never, discussed. Postoperative follow-up may be regarded as the screening of a high-risk group. The reduced mortality seen in the meta-analysis of randomised controlled trials is here regarded as small and should also be weighed against the drawbacks of follow-up before any recommendation for more active surveillance is given. PMID- 14558211 TI - [New studies of COX-inhibitors, yet issues remain]. AB - Advantages and risks related to the use of selective COX-2 inhibitors when treating arthritis are currently being scrutinized by authorities and public. The discussion tends towards exaggerated claims for or against their usefulness. The issue of cardiovascular safety is still not finally settled. In an experimental study using patients with severe coronary disease, administration of celecoxib resulted in improved endothelial function together with reduced CRP levels. Gastrointestinal tolerance was studied in patients who had recently recovered from peptic ulcer bleeding. In this group of high risk patients, celecoxib was as safe as combined therapy using omeprazol and diclofenac when given for 6 months. However, both COX inhibitors caused hypertension and adverse renal effects. The second generation of selective inhibitors is being launched. Etoricoxib--related to rofecoxib--was shown to be as potent as indomethacin in the treatment of acute gout, but it caused fewer adverse reactions. In general, however, any advantage of second generation as compared to first generation COX-2 inhibitors remains to be proven. The Swedish Council on Technology Assessment in Health Care, in its "SBU Alert", has published an appraisal of celecoxib and rofecoxib, in which the need for further long-term safety studies is emphasized. PMID- 14558212 TI - [Fear of childbirth a challenge in health care. Cesarean section not the only way out, professional support during pregnancy and planned delivery a relief to many]. PMID- 14558213 TI - [Pseudomyxoma peritonei--a syndrome with varying prognosis]. AB - Pseudomyxoma peritonei (PMP) is an unusual condition in which massive amounts of gelatinous fluid collections are associated with mucinous implants on the peritoneal and omental surfaces. Recent molecular genetic studies suggest the appendix as the most common primary origin of this disease. Surgical debulking in a combination with intraperitoneal (i.p.) chemotherapy has been advocated to treat this disease. Thirty patients were treated for PMP, by this concept, at Uppsala University hospital between 1993 and April 2003. One patient died because of septic complications (inhospital mortality 3 per cent). Twenty-nine patients are alive after median follow-up of 16 months (range 1-92), 18 of them tumor free. PMID- 14558214 TI - [Closing-down of a palliative unit. You are a county chief physician]. PMID- 14558215 TI - [The painter Francis Bacon: "We are born and we die, and in between we try to give meaning to this meaningless existence by our driving forces"]. PMID- 14558216 TI - [Give more status to the clinical conversation within child and adolescent psychiatry]. PMID- 14558217 TI - [Sufficient medical safety at the Arlanda airport]. PMID- 14558218 TI - [Too pessimistic view on screening and follow-up of colorectal cancer]. PMID- 14558219 TI - [RELIS--a Norwegian database with free, problem-based drug information for health care]. PMID- 14558220 TI - [A patient received a drug which was not the prescribed one]. PMID- 14558221 TI - Pioneering group practices explore new technologies. PMID- 14558222 TI - Physicians vocalize dictation issues. PMID- 14558223 TI - In paper war, is OCR a good soldier? PMID- 14558225 TI - Will health care get smart? PMID- 14558224 TI - Getting claims paid in a HIPAA world. PMID- 14558226 TI - Let your fingers do the browsing. PMID- 14558227 TI - Medical records, I.T. lines blurring. PMID- 14558228 TI - Provider begins carding patients. PMID- 14558229 TI - The human genome, gerontological nursing, and cautious optimism. PMID- 14558230 TI - Nursing indicators of quality in nursing homes. A Web-based approach. AB - This article is an examination of websites providing consumer information about nursing home quality of care, including existing federal and state websites and a new comprehensive website designed for California nursing homes. The article focuses on research and information related to nursing indicators of quality used for the California nursing home website. It includes staffing levels (e.g., hours, types, turnover rates), financial indicators (e.g., direct care expenditures, wages, benefits), and complaints and deficiencies. Overall, nursing indicators of quality are a major approach for evaluating nursing home quality and can be used by nurses, consumers, and advocates. PMID- 14558231 TI - Institutionalization in Taiwan. The role of caregiver gender. AB - The role of caregiver gender in the likelihood of institutionalization of Taiwanese older adults was explored in this study. A sample of 78 male and 69 female primary caregivers of elderly patients who had experienced a stroke at least 6 months prior to the study were interviewed. Logistic regression analyses were applied to examine direct and interaction effects of the elderly adult's functioning the caregiver's available resources, the degree of caregiver burden, perceived public opinion toward institutionalization, and precipitating events on the likelihood of institutionalization among Taiwanese male and female caregivers. Women were more likely to institutionalize the older adult for whom they cared. The proposed model correctly predicted the likelihood of institutionalization of an elderly adult based on male versus female caregivers at the 92% level. Perceived public opinion toward institutionalization was the most significant predictor of institutionalization for both genders. Perceived public opinion toward institutionalization has a strong influence on whether or not caregivers institutionalize an elderly relative. This is consistent with Chinese culture in which public opinion has a much stronger effect on individual behavior than in the United States. American concepts of "minding one's own business" do not exist in Taiwan. It is logical that the older adults' level of functioning would predict the likelihood of institutionalization regardless of caregiver gender. In terms of caregiver characteristics, working hours in male caregivers is more predictive, and the quality of the relationship with the older adult was more predictive of institutionalization for female caregivers. PMID- 14558232 TI - Alcohol use in a continuing care retirement community. AB - The purpose of this study was to explore alcohol use among older adults living in a continuing care retirement community (CCRC) and to consider differences in overall health, number of chronic illnesses, falls, age, and exercise behavior between those who drink moderately versus those who do not. A one-time face-to face interview was conducted with a total of 316 residents living in an urban CCRC in the northeast. The participants were 86.6 6 6.3 years of age, and the majority were women (79%), unmarried (75%), White (9%), and middle to upper socioeconomic status (100%). The majority of these individuals (63%) drank moderately, consuming at least one drink daily. No difference was found between men's and women's drinking behaviors (chi 2 = .47, p > .05). Adults who drank moderately were older (F = 8.7, p < .05), more likely to exercise regularly (chi 2 = 27, p < .05), and were more likely to have had a fall (F = 3.7, p < .05). No statistically significant difference was found between the groups related to perceptions of overall health, number of chronic illnesses, number of medications, overall or number of medications that interact with alcohol, or cognitive status. Although not statistically significant, a greater percentage of individuals who drink moderately had fractures (12%) when compared to the percentage of individuals who do not drink at least moderately (7%). Recognizing both benefits and risks to alcohol use in older adults suggests that an individual approach to educating these individuals about their use of alcohol should be conducted. PMID- 14558233 TI - Bladder management in adult care homes. Review of a program in North Carolina. AB - In North Carolina there are approximately 34,000 residents in adult care homes (ACHs). Approximately 40% of these residents have urinary incontinence, and others require assistance with toileting. High prevalence of cognitive impairment, few licensed staff, and low staff-to-resident ratios in ACHs make behavioral techniques used in community-dwelling populations and toileting programs used in nursing homes inappropriate for these residents. This program was implemented using a two-level approach (facility and individual resident) and uses an education consultation approach for implementation. PMID- 14558235 TI - Family and staff partnerships in long-term care. A review of the literature. AB - Working with the family in aged care to provide the best care possible is consistent with modern nursing philosophy, which espouses holistic care. The quality and enjoyment of the experience, however, is frequently fraught with problems and challenges for both the family and the staff involved. In residential aged care, partnerships are a complex mix of interactions among the older adult resident, the family, and the caregiving staff. To understand how family involvement in this environment can be made more meaningful, nurses need to be cognizant of how family members experience the caregiving role and how they are perceived by nursing home staff and residents. The relevant literature of the past 20 years is reviewed and key issues central to an understanding of "family care" in the nursing home from the perspective of the family, the nursing home resident, and the nursing home staff are highlighted in this article. PMID- 14558236 TI - [Evidence Based Medicine, in Revista Medica de Chile]. AB - Since the beginning of the 20th century, the principles of the experimental method in biological sciences have been progressively applied in biomedical and clinical research. After the pioneering work of AB Hill et al, biostatistics became a fundamental tool in the appraisal of new therapeutic agents or methods, and of diagnostic tests. From 1990 on, "Evidence based Medicine" has been added to it. This journal opens in this issue a section containing critical analysis of articles published in the medical literature deserving the attention of its readers. However, as several other medical journals have stressed, we recommend that the conclusions of these analysis should not be used as a substitute for clinical judgment and they do not represent an official position of our journal. Revista Medica de Chile welcomes the new Section and invites contributions to it. PMID- 14558234 TI - Behaviors of older men living in the community. Correlates producing active composure. AB - The correlates of active composure behaviors for older veteran men living in the community were investigated to identify this aspect of the complex domain of health behavior. Cox's (1982) Interaction Model of Client Health Behavior was used to frame this investigation. The independent variables were age, education, race, marital status, children, siblings, income, spiritual well-being, functional status, motivation, health conceptions, and loneliness. The dependent variable was Schwirian's (1992) active composure, conceptualized as activities producing rest, relaxation, and anxiety and stress reduction. The sample was male veterans (N = 184) older than 65 who were not living in an institution. A multiple regression model explained 49% of the variance in active composure. Race, income, religious aspect of spiritual well-being, instrumental activities of daily living, and loneliness were significant predictors. Results suggest that nurses may find these correlates helpful in facilitating the identification of risk status and developing active composure self-care strategies for older men living in the community. PMID- 14558237 TI - [Biochemical phenotypes and phage types of Salmonella enteriditis strains isolated in Antofagasta during the period 1997-2000]. AB - BACKGROUND: PhP-S48 (Phene Plate Techniques AB), a method based on biochemical phenotypes has been developed and used successfully to typify S enteritidis strains in epidemiological studies. AIM: To identify phenotypes of S enteritidis isolated from eggs, chicken meat and infected humans in Antofagasta during the period 1997-2000. MATERIAL AND METHODS: PhP-S48 and phage typing were used to identify phenotypes of 33 S enteritidis strains, sixteen isolated from poultry and 17 from clinical sources. S enteritidis ATCC17036 was used as control strain. RESULTS: Twelve biochemical phenotypes (BTs) including 4 common (C) and 8 single (S) were identified. BTs C1 y C3 containing 16 and 5 strains, respectively, accounted for 63.6% of the isolates. BT C1 was found in poultry and human sources in the period 1997-2000, and BT C3 was isolated from humans, in the period 1999 2000. Using phage typing, 5 phage types (PT) and 3 strains could be not typed (NTs). PT1 and PT21 were the dominant phage types, with 14 and 13 strains respectively. Strains of PT1 were isolated from poultry and human sources in the period 1997-2000. PT21 was found in poultry samples in the period 1997-1998 and in clinical samples, in the period 1997-1998. Combination of biochemical phenotypes and phage typing divided the strains into 5 phenotypes (BT:PT). Two phenotypes were the most frequently isolated, phenotype C1:1 with 8 isolates found in eggs and humans in 1999, and phenotype C1:21 with 5 strains isolated in 1997-1999. CONCLUSIONS: These results indicate the presence of one persistent and one recently emerged phenotype among S enteritidis in Antofagasta, Chile. PhP-S48 also provided information about a relationship among the strains. PMID- 14558238 TI - [Medical outcomes and antimicrobial compliance according to the Chilean Society of Respiratory Diseases guidelines for hospitalized patients with community acquired pneumonia]. AB - BACKGROUND: The Chilean Society of Respiratory Diseases (SER) developed guidelines for the empirical treatment of community acquired pneumonia (CAP). AIM: To evaluate the degree of adherence to antibiotic treatment recommended by SER guidelines and its influence on medical outcomes. PATIENTS AND METHODS: We prospectively evaluated 453 consecutive immunocompetent adults (mean age +/- SD: 69 +/- 19 years) hospitalized for CAP. Patients were stratified according to the Pneumonia Severity Index (PSI), and initial antibiotic regimen was classified as being consistent or inconsistent with the SER guidelines. Rate of medical complications, switch therapy rate, length of stay (LOS), and 30 days mortality were compared between those treated consistently or inconsistently with the SER guidelines. RESULTS: Adherence to SER guidelines was 46%. Patients treated consistently with the SER guidelines were older (mean age +/- SD: 72 +/- 16 v/s 65 +/- 20 years), had more comorbidities (84 v/s 69%) and a higher proportion belonged to the high-risk PSI categories (69 v/s 49%). There were no significant differences in medical complication rate, switch therapy rate or LOS between both groups. Adherence to SER guidelines did not affect mortality after adjusting for PSI and for prognostic factors associated with 30 days mortality by multivariate analysis. CONCLUSIONS: The degree of adherence to antibiotic treatment recommended by SER guidelines was moderate and they were applied mainly in patients with high risk CAP. This fact can explain the lack of evidence of improved medical outcome in patients treated according to SER guidelines. PMID- 14558239 TI - [Results of a multidisciplinary and intensified treatment program for type 1 Diabetes Mellitus in a Chilean Public Hospital]. AB - BACKGROUND: During the last decade, the importance of glycemic control in the prevention of the microvascular complications of type 1 Diabetes Mellitus (DM1) was clearly demonstrated. AIM: To evaluate the metabolic and anthropometric results of a multidisciplinary intensified treatment program of DMI in children and adolescents. PATIENTS AND METHODS: Report of 54 patients treated during 2001. The intensified treatment consisted of: multiple daily doses of insulin, frequent glycemic control, nutritional, psychological and educational support, and permanent availability of a diabetes nurse for telephonic support. RESULTS: Thirty one patients were female, their mean age was 10.4 +/- 0.5 years old and 52% were experiencing puberty. Fifty three percent of the patients used 3 insulin doses per day, 95% changed rapid insulin dose based on glucose levels and 18% considered carbohydrates in their rapid insulin dosing. Mean glycosilated hemoglobin was 8.18 +/- 0.23% without differences by sex or pubertal status. Sex, pubertal stage and the number of insulin doses did not contribute to glycosilated hemoglobin changes. There were no differences in weight or BMI, but there was a decrease in height Z score from the admission to the program until the last control (0.1 +/- 0.1 vs--0.3 +/- 0.1 DS; p < 0.01). CONCLUSIONS: A modified intensified modality of DM1 therapy for pediatric patients in a public hospital in Chile is feasible, achieving similar metabolic control, compared to international large centers. PMID- 14558240 TI - [Bovine serum albumin (BSA) antibodies in children with recently diagnosed type 1 diabetes with breast feeding and milk exposition]. AB - BACKGROUND: Environmental and genetic factors (viruses, toxins and diet) are involved in the aetiology of type 1 diabetes. Among the dietary factors, the role of breast feeding and exposure to cow's milk proteins deserve special attention. AIM: To determine the anti-BSA-IgG levels in type 1 diabetic children and to analyse the possible association with breast feeding duration, exposure to cow's milk and beta pancreatic auto-antibodies. PATIENTS AND METHODS: Blood samples were collected from 161 diabetic children and 144 controls to measure anti-BSA IgG level, GAD65, IA-2 and ICA autoantibodies. All children answered a questionnaire about dietary habits during infancy. RESULTS: anti-BSA-IgG was positive (using a cut off point of 25.6 ng/ml) in 98% of diabetic children and 0% of the control population. The length of breast feeding or early exposure to cow's milk did not influence the concentration of anti-BSA-IgG. Positive BSA titers did not increase the beta pancreatic reactivity (ICA+, GAD+, IA2+). CONCLUSIONS: Our data confirm the high frequency of anti-BSA-IgG among diabetic children. However, a specific role in the immunological process of type 1 diabetes cannot be attributed to this protein. PMID- 14558241 TI - [Tobacco smoking and mental health]. AB - BACKGROUND: Smoking continues to be one of the most important health burdens worldwide. AIM: To describe smoking habits and associated risk factors in the population of Santiago, Chile. MATERIAL AND METHODS: A cross sectional study of a representative sample of the population, from 16 to 64 years old, residents of Santiago, Chile (total population: 3,237,286). A structured interview that included questions about use of tobacco, the CIS-R interviews, used for common mental disorders, were applied. RESULTS: From the sample of 4,693 households, 3,870 people were interviewed (52.2% women, 47.8% men) and 10% refused. Forty percent of the population currently smoked (52.5% men, 47.8% women). "Being a current smoker" was associated with being younger than 55, male sex, and having a common mental disorder. DISCUSSION: Smoking is highly prevalent in Chile, as compared with developed countries and with some developing countries. Gender differences in use of tobacco have decreased. A higher risk of smoking for people with mental disorders is confirmed. PMID- 14558242 TI - [Clinical study of measure of parasitemia in patients infected with Trypanosoma cruzi in Atacama, Chile]. AB - BACKGROUND: Trypanosoma cruzi infection is endemic in Northern/Central Chile. AIM: To perform a clinical assessment of patients infected with Trypanosoma cruzi. PATIENTS AND METHODS: Two hundred sixty three subjects with a positive serology for Trypanosoma cruzi, were invited by mail to a clinical assessment in a Regional Hospital. In a subsample of these, a polymerase chain reaction for Trypanosoma cruzi, was done. RESULTS: Of all the invited subjects, 183 responded and were assessed at the hospital. Of these, 60 had cardiac affections, 52 had colon problems and 17, esophageal disease. Seventy four were asymptomatic. Of the 64 patients in whom polymerase chain reaction was done, 35 had a positive result. CONCLUSIONS: A high percentage of subjects infected with Trypanosoma cruzi, had clinical consequences of the infection. Polymerase chain reaction showed persistency of the parasite in more than half of the infected patients. PMID- 14558244 TI - [Geriatric assessment in the primary care]. AB - BACKGROUND: A good medical care program for the elderly requires a thorough assessment of the functional capacity and limitations of the consulting patients. AIM: To perform a geriatric assessment to elderly subjects consulting in a public outpatient clinic. PATIENTS AND METHODS: A cross sectional assessment of subjects over 60 years old, beneficiaries of the public health care system and consulting in 4 outpatient clinics of a poor community of Metropolitan Santiago. Evaluation included a complete medical history, the Katz score for activities of daily living (ADL), Lawton score for instrumental activities for daily living (IADL) and Folstein Mini Mental state examination. RESULTS: During the study period 1,023 subjects, 73% female, aged 74 +/- 7 years, were assessed. Thirteen percent were illiterate, 78% elementary school, 8% had high school and 0.2% university education. Eighty eight percent lived with relatives, 79% did not participate in community groups and 7% smoked. The most common chronic disease detected was high blood pressure in 61%, followed by obesity in 32%, osteoarthritis in 22%, diabetes mellitus in 17%, depression in 11%, cardiac diseases in 8%, hypothyroidism in 3% and epilepsy in 1%. ADL score showed partial dependency in 11% and total dependency in 2%. IADL score showed partial dependency in 41% and total dependency in 4%. Mini Mental score was abnormal in 31% of subjects. CONCLUSIONS: High blood pressure, osteoarthritis, depression and cognitive impairment are the most common health problems in this sample of elderly subjects. ADL score showed a low rate of dependency. PMID- 14558243 TI - [Falls in institutionalized elderly subjects. Features and geriatric assessment]. AB - BACKGROUND: Falls are an important cause of disability and mortality among elderly subjects. AIM: To study the features and incidence of falls in institutionalized elderly subjects. PATIENTS AND METHODS: Prospective recording of all falls, occurring in a period of eight months, to 453 subjects older than 60 years, living a home for the elderly. Evaluation of functional status of subjects suffering falls, using the Spanish Red Cross score. One hundred and two subjects living in the same place, but not suffering falls, were considered as controls. RESULTS: One hundred and three subjects, aged 80 +/- 6 years (24% of the sample) suffered falls. Most falls were during the day and while walking. Seventy percent of subjects suffering falls did not have a history of previous falls. Nine percent of those falling, suffered a fracture. Compared to controls, falling subjects had a more deteriorated functional and mental status and consumed more benzodiazepines and neuroleptics. CONCLUSIONS: There is an association between functional status and psychotropic medication consumption and the incidence of falls in institutionalized elderly subjects. PMID- 14558245 TI - [The influence of winter 2002 in pediatric health: dissociation between environmental factors and respiratory syncytial viruses, in Santiago]. AB - BACKGROUND: The increase in winter related health pediatric demand is associated with three factors: cold and rainy weather, air pollution and respiratory viral epidemics. During the winter of 2002 there was, successively, heavy rain, air pollution, cold weather and a respiratory syncytial virus (RSV) epidemic in Santiago. AIM: To study the influence of environmental factors and RSV epidemic on pediatric health care demand. PATIENTS AND METHODS: The number of hospital admissions and outpatient consultations for acute lower respiratory infections (ALRI) from April to October 2002 (Autumn to Spring in Chile), were registered in a public pediatric hospital of Santiago. A respiratory virus surveillance (RSV, adenovirus influenza and parainfluenza virus) was carried out among children admitted for ALRI. Climate conditions and air pollution (number of particles of more than 10 microns/mm3) values were provided by local environmental health services. RESULTS: As expected, a rise in winter hospital admissions and outpatient consultations was detected, that peaked in week 29. This rise coincided with the higher RSV detection week. There was heavy rain in weeks 22, 23 and 30, as well as cold weather in June-July (weeks 23, 24, 26, 27 and 30; mid Winter), that did not increase health care demand. Likewise, high air pollution, registered from weeks 24 to 26, did not increase health care demand. CONCLUSIONS: RSV epidemic is the principal factor associated to the increase in health burden during winter in Santiago. PMID- 14558246 TI - [Reversion of hypophosphatemia after the excision of a composite hemangioendothelioma in the great toe]. AB - We report a 43 years old male admitted to the hospital for progressive lumbar pain, lasting 20 years, that caused severe disability. On admission the patient had a serum phosphate of 2 mg/dl, an urine phosphate excretion over 300 mg/dl and serum alkaline phosphatases over 750 U/L. Serum intact parathormone was normal and tubular maximum phosphorus/glomerular filtration was 0.7 mg/dl. Bone scintigraphy showed an increased radionuclide uptake in condro-costal joints. Bone densitometry showed femoral osteoporosis. A violet colored mass was detected in a great toe. It was removed and the pathological diagnosis was a composite hemangioendothelioma. After tumor excision, serum phosphate levels returned to normal values and symptoms disappeared within 15 days. PMID- 14558247 TI - [Dementia caused by vitamin B12 deficiency. Clinical case]. AB - Cyanocobalamin (vitamin B12) deficiency can cause polyneuropathy, myelopathy, blindness, confusion, psychosis and dementia. Nonetheless, its deficiency as the sole cause of dementia is infrequent. We report a 59 years old man with a 6 months history of progressive loss of memory, disorientation, apathy, paranoid delusions, gait difficulties with falls, and urinary incontinence. He had suffered a similar episode 3 years before, with a complete remission. On examination there was frontal type dementia with Korsakoff syndrome, a decrease in propioception and ataxic gait. Cerebrospinal fluid examination showed a protein of 0.42 g/L. Brain computed tomography showed sequelae of a frontal left trauma. Brain single photon computed tomography (SPECT) was normal. Complete blood count showed a macrocytic anemia with a hematocrit 29% and a mean corpuscular volume of 117 micron3. Plasma vitamin B12 levels were undetectable, erythrocyte folate levels were 3.9 ng/ml and plasma folate was normal. The myelogram showed megaloblastosis and the gastric biopsy showed atrophic gastritis. Treatment with parenteral B12 vitamin and folic acid reverted the symptoms, with normalization of the neuropsychological tests and reintegration to work. PMID- 14558248 TI - [Rhomboencephalitis caused by listeria monocytogenes infection in the central nervous system]. AB - Listeria monocytogenes infections can involve the central nervous system in the form of a rhomboencephalitis. Three possible cases of rhomboencephalitis by Listeria monocytogenes are reported (2 females, aged 44 and 49 and a man of 36 years old). The three cases were preceded by an unspecific prodrome of headache, vertigo and fever in absence of a meningeal syndrome. The neurological stage was defined by the unilateral involvement of cranial nerves and the cerebellum and a clear inflammatory cerebrospinal fluid (CSF) with the presence of polymorphonuclear leukocytes, and normal glucose and protein levels. A magnetic nuclear resonance (MRI) showed the appearance of characteristic images, present in the bulboprotuberancial region. These images are one of the most constant features of this disease, reported in the literature. The early diagnosis of rhomboencephalitis was based on the clinical picture, the study of the CSF and the MRI, allowing the use of antimicrobials, prior to microbiological identification. Therefore, the risk of brain stem and cardiac complications of the disease is reduced. PMID- 14558250 TI - [Masters of Chilean Internal Medicine]. AB - Master of internal Medicine is the highest award granted by the Chilean Society of Internal Medicine (Sociedad Medica de Santiago) to its members. This article outlines the sense and transcendence of this award and provides an abridged, certainly incomplete, data of the recipients since 1981 when this award was first given. Those who have received it are not the only ones that have the attributes to deserve such distinction. There are many members with similar or even better merits, and therefore the Directory of the Society is usually exposed to a very hard decision when facing each new nomination. When a member is awarded, he becomes a model to be emulated or surpassed by the future generations; thus assumes the great responsibility of maintaining the spirit, the prestige and the projection of Internal Medicine into the future. PMID- 14558249 TI - [Abraham Horwitz (1910-2000): a Leading Man of Pan American Public Health]. AB - The Chilean physician Abraham B Horwitz (1910-2000) was an outstanding personality of World and Pan American public health during the second half of the twentieth century. He was member of a family that, emigrating due to ethnic persecutions in Russia, took refuge in Chile. He became a relevant physician, a specialist in infectious diseases and public health. He was highly influential in the birth of the Public Health School at the University of Chile and the Chilean Health Service. He became Executive Director of the Pan American Health Organization, holding that position for 16 years. During this period, the institution experienced a great development. He stimulated research in the areas of basic sanitation, nutrition and eradication of transmissible diseases. He also opened unexplored areas such as the relation between economy, modern administration and health. During his last years, he chaired a successful Nutrition Committee at the United Nations. The most outstanding achievement of this Committee was the promotion of widespread vitamin A use. His intellectual and social deed is continued by The Pan American Foundation for Health and Education and this institution established an annual prize in his memory. PMID- 14558251 TI - [Evidence based medicine: a view after a decade]. AB - Evidence based medicine (EBM) appeared early in the 1990s and since then it has been developed and expanded worldwide. A decade later we summarize the history of EBM, the initial debates and the evolution to the current concept of evidence based health care (EBHC) as a tool for clinical decision making. We also describe the process of EBHC, some insights to current dilemmas and the situation of EBM in Chile. PMID- 14558253 TI - Surgical management of ascending aortic disease. AB - Two cases of aortic pathology have been presented--namely, ascending aortic aneurysm and ascending aortic dissection. These entities are distinct, but can be superimposed. Surgical resection is the procedure of choice for both these entities, in contradistinction to descending aortic dissections. Although dealing with these entities can indeed be a humbling experience, lifesaving treatment can be delivered by a combination of a high index of suspicion, early diagnosis, and prompt surgical therapy. PMID- 14558254 TI - Managing risk in a risky world. PMID- 14558255 TI - The good doctor of Blue Mountain--Jessie Mauney, MD. PMID- 14558256 TI - Looking back: cardiac stimulants. 1911. PMID- 14558257 TI - Information and quality healthcare-expansion of Medicare beneficiary protection. PMID- 14558258 TI - [Why did they die? Consequences of a heatwave]. PMID- 14558259 TI - [Gaucher's disease: a second drug is now available]. PMID- 14558260 TI - [Dengue fever: outbreak in southern Europe?]. AB - Epidemiology of dengue fever is changing dramatically. The worldwide incidence is rising and clinical symptoms are worsening. Reports describing forms associated with haemorrhages or shock syndrome involving both children and adults are increasingly frequent in regions beyond Southeast Asia where the first cases were observed. Many mechanisms could be implicated in these changes, including modifications of the virus, host, vector, or socio-economic factors. The new facilities in the laboratory diagnostic (MAC-ELISA; molecular biology), the commercialization of these assays allow not only assessment of morbidity and mortality in endemic areas and early detection of epidemic outbreaks but also evaluation of socio-economic impact and effectiveness of control measures. Now, the efficiency of the fight must be better, otherwise dengue will grow up during this new century. PMID- 14558261 TI - [Community acquired pneumonia: higher mortality than traffic accidents ]. PMID- 14558262 TI - [Diagnosis of community acquired pneumonia in adults]. AB - In contrast with acute bronchitis, which is benign and very frequent, community acute pneumonia has to be considered as a serious illness with a mortality rate that can reach 10-15% in inpatients. In adult, pneumonia is usually due to bacteria and antibiotherapy is always required. Clinical diagnosis is based on presumption signs: fever, tachycardia, tachypnea, thoracic pain, localised rales and a global severity impression. A chest X-ray is required. The pathogens involved are usually Streptococcus pneumoniae or atypical organisms. In inpatients (severe pneumonia or presence of risk factors), other pathogens such as gram negative bacilli or anaerobes are possible. The accurate diagnosis is based on invasive methods, which are not justified in outpatients but indicated in severe pneumonia. PMID- 14558263 TI - [Community acquired pneumonia in children]. AB - In children, community acquired pneumonia represent less than 10% of respiratory tract infections. In infants and toddlers, the diagnosis is difficult as, at any age, in case of isolated fever. Typical features of pneumonia are rare. Chest radiograph is necessary for confirmation and mandatory in children under 2 years of age, other diagnosis suspision or recurrent pneumonia. Identification of risk factors and/or severity criteria will guide the hospitalisation decision. Epidemiological data have a collective usefulness but help very little for the individual decision. Pneumococcus is observed at any age. Before 3 years of age, viruses and pneumococcus are predominant. After 3 years of age pneumococcus, Mycoplasma pneumoniae and Chlamydia pneumoniae are possible. Viral or virobacterial co-infections exist in 15 up to 25% of cases. An empiric and rapid antibiotherapy is given, pending on age, national or regional epidemiology, initial severity, risk factors. A clinical evaluation is mandatory at H48-72. Routine vaccination with the 7-valent conjugate anti-pneumococcal vaccine will modify the strategy of use of antibiotics in children's pneumonia. PMID- 14558264 TI - [Community acquired pneumonia in the elderly]. AB - Pneumonia, more frequent in the elderly, results in higher morbidity and mortality, and is a frequent cause of hospitalisation. Although the incidence of pneumonia increases with age, mainly after 70-75 years, life in institution, comorbid medical illness (heart disease, chronic obstructive pulmonary disease, dementia), malnutrition and defence impairments are independent risk factors. Older patients with pneumonia complain of significantly fewer symptoms than younger patients. Streptococcus pneumonia is the most common cause of pneumonia, atypical pathogen are rare. There is a shift toward gram-negative bacteria and opportunistic flora with increasing age and severity of concomitant medical illness. Anaerobic bacteria are frequent in aspiration pneumonia. To hospitalize or not is the first decision to take, based on clinical criteria and risks factors. Use of the guidelines for empirical treatment of pneumonia is recommended. Old patients often recover slowly. Pneumococcal and influenza vaccinations are effective and warranted. PMID- 14558265 TI - [Influenza pneumonia]. AB - Two types of pneumonia are well recognized during influenza: primary viral pneumonia and secondary bacterial pneumonia. Primary viral pneumonia occurs after a typical onset of influenza with rapid progression of dyspnea and cough leading to acute respiratory distress syndrome. Treatment consists of respiratory assistance, but mortality is high. Secondary bacterial pneumonia occurs more frequently in the elderly and in patients with chronic pulmonary diseases. Staphylococcus aureus, Streptococcus pneumoniae and Haemophilus influenzae are the most frequently isolated bacteria. After an initial phase of clinical improvement, manifestations of bacterial infection with pulmonary consolidation occur. The outcome is favorable with antibiotics but depends on the patient's underlying conditions. PMID- 14558268 TI - [Searching community acquired pneumonia in MEDLINE]. PMID- 14558266 TI - [Diagnosis of legionellosis]. AB - The majority of legionellosis are caused by Legionella pneumophila, especially serogroup 1, the only diagnosed by Legionella urinary antigen test. The diagnosis of other serogroups and other Legionella species is based on culture and seroconversion. Sever cases are more frequently observed in immunocompromised patients and in nosocomial pneumonias. Each case must lead to look for the source of contamination, to detect other potential patients and permit environmental measures. Clinically and radiographically, the pneumonia is non specific except if suggestive gastro-enteric or neurologic symptoms are present. The diagnosis could be suspected in case of relative bradypnia or beta-lactamin resistance. PMID- 14558267 TI - [Treatment and prevention of pneumococcal pneumonia]. AB - Streptococcus pneumoniae is the most commonly identified pathogen in patients with community-acquired pneumonia. 40% of isolated strains in France are of decreased sensibility to penicillin, two third being multiresistant to antibiotics. However, high doses of some beta-lactams are effective in vivo against the majority of circulating strains (MIC > 2 mg/L). For this reason according to French guidelines amoxicillin (3 g/day) is the first line recommended treatment. Telithromycin is an alternative, or ceftriaxone (1 g/day) in more severe cases. If the level of resistance increases (MIC > 4 mg/L) guidelines would be revisited. Effectiveness of pneumococcal vaccination has been confirmed in cases of bacteriemic pneumococcal pneumonia (elderly patients included). Vaccine is recommended among persons with comorbiditie(s) and 65 years old population. PMID- 14558269 TI - [Reactions]. PMID- 14558270 TI - [Type II diabetes in children and adults]. PMID- 14558271 TI - [Acute abdominal and back pain in adults]. PMID- 14558272 TI - [HIV infection]. PMID- 14558273 TI - [Acute and chronic dyspnea]. PMID- 14558275 TI - Study shows varied responses for burning mouth syndrome. PMID- 14558274 TI - [Jean Comandon and the beginning of microcinematography]. PMID- 14558276 TI - Insurance audits: what to do. PMID- 14558278 TI - Donated dental services: needed now more than ever. PMID- 14558277 TI - How to handle an angry patient. PMID- 14558279 TI - Case study: the doc who wouldn't help an HIV patient. PMID- 14558280 TI - How to safeguard the value of your practice. PMID- 14558281 TI - [Portable ultrasound devices. A new era for clinical cardiologists]. AB - Recent advances in ultrasound technology have led to the development of fully portable ultrasound machines for point-of-care cardiac examination. Limited comparative studies indicate that these devices are more accurate than physical examination in identifying cardiac abnormalities. Though their diagnostic performance is generally inferior to standard echocardiography, there appears to be good concordance at least for some conditions. However, it must be emphasized that reported results were obtained with cardiologists or sonographers trained and experienced in performing and interpreting echocardiography. The potential role of these devices in cardiac care will depend on their impact on patient management, compatibility with individual practice, their cost, and further technical developments. Operator training in performance and interpretation of examinations obtained using these ultrasound devices seems to be vitally important. PMID- 14558282 TI - [Portable ultrasound devices in clinical cardiology: lights and shadows]. PMID- 14558283 TI - [Hormone replacement therapy and cardiovascular prevention: many statements, some doubts]. AB - Results of randomized, placebo-controlled clinical trials with major clinical endpoints as well as with angiographic or echographic endpoints recently completed with hormone replacement therapy in postmenopausal women, are critically discussed. These results, far from confirming the protective cardiovascular effect previously found in observational studies, showed that hormone replacement therapy is associated with an increase of coronary risk, mainly in the first year of treatment, of stroke, as well as of venous thromboembolism and invasive breast cancer. Various plausible explanations, both from the methodological and the biological point of view, for the divergent findings between the randomized clinical trials and the observational studies are critically examined. On the basis of the available data, no cardiovascular preventive role must be assigned to the hormone replacement therapy; the only today accepted indication remains the short-term treatment of postmenopausal vasomotor disturbances. PMID- 14558285 TI - [Normal and abnormal atrial activation. An update with special reference to surface electrocardiogram]. AB - Surface electrocardiogram is commonly used to evaluate the normal and abnormal activation of the atria. Interatrial conduction appears as the most important factor determining P wave duration and morphology during sinus rhythm. A good estimation of the interatrial conduction time can be obtained by a simple esophageal recording or by P wave duration on the surface electrocardiogram. A careful analysis of the ectopic atrial beats on the 12-lead electrocardiogram can identify the site of origin of the ectopic atrial beat. Surface electrocardiogram is commonly used in the classification of atrial flutter. PMID- 14558284 TI - [Biology and physiopathology of tissue factor and its relevance for cardiovascular diseases]. AB - Tissue factor (TF) is a transmembrane glycoprotein, currently considered as being the major regulator of the coagulation cascade and the initiator of thrombogenesis in vivo. When TF comes in contact with blood, it forms a high affinity complex with factors VII/VIIa, activating factors IX and X and thus leading to the formation of an insoluble fibrin clot. The regulation of TF-VIIa activity plays a key role in blood-vessel wall interactions. Selective patterns of cellular expression of TF are observed in tissues. TF is constitutively localized only on the surface of cells anatomically separated from the blood, where it plays an essential role in hemostasis by limiting hemorrhage after vessel wall injury. A number of pathophysiologic stimuli are capable of inducing TF transcription and activity in endothelial cells and monocytes. An aberrant TF expression in contact with blood is implicated in thrombotic complications of atherosclerosis, including acute myocardial infarction. Recent findings have demonstrated cell-derived microparticles containing TF in the circulating blood of patients with acute coronary syndromes, capable of triggering and propagating thrombus growth. This observation suggests a new view of thrombosis that does not necessarily require the exposure of vessel wall-derived TF at the site of vascular injury to initiate and propagate thrombosis. PMID- 14558286 TI - [Temporary emergency cardiac pacing under continuous echocardiographic control. Feasibility and safety of the procedure without using fluoroscopy]. AB - BACKGROUND: Cardiac pacing often turns out to be the only effective treatment of severe brady-arrhythmias. Several invasive and noninvasive temporary pacing procedures are known, whose application is sometimes difficult or time-consuming. An alternative temporary cardiac pacing procedure is described in this article, which is based on echocardiographic control. METHODS: Fifty-four nonconsecutive patients were studied; they all were needing urgent cardiac pacing. A first choice attempt to perform an ultrasound-guided temporary cardiac pacing, by using a right jugular venous approach, was done. The catheter pathway was monitored by means of echocardiography performed by another operator. All data concerning time of execution, pacing parameters, acute or chronic complications and the in situ time duration of the catheter without needing to be repositioned were obtained. RESULTS: Ultrasound-guided cardiac pacing was not feasible in 3 patients (6%), because of a high thoracic acoustic impedance or failing to perform right jugular venous catheterization. Mean execution time was 680 +/- 179 s. Echocardiographic monitoring was performed in 50 patients by a cardiologist, in 1 patient by a non cardiologist physician, and in 3 patients by a nurse, who had been previously trained to keep the transducer in the right position. No serious complications occurred. On 4 occasions (8%) the catheter had to be repositioned during the following hours. On 25 occasions (46%) permanent cardiac pacing was finally performed. CONCLUSIONS: Temporary ultrasound-guided cardiac pacing seems to be a safe and easy procedure; it can be performed in a broad range of clinical and logistic scenarios and does not require fluoroscopic monitoring. A limited number of human resources is needed, but technical skills in central venous catheterization are required. Further studies are needed to validate this procedure; however it shows several potential benefits as compared to the other temporary cardiac pacing techniques. PMID- 14558287 TI - [Non-invasive 16-row spiral multislice computed tomography coronary angiography after one year of experience]. AB - The gold standard for direct diagnostic imaging of coronary arteries is still conventional X-ray coronary angiography, even though for several other applications, noninvasive techniques substituted invasive ones. In the last 10 years several techniques (magnetic resonance, electron beam tomography, spiral computed tomography) attempted to emerge as noninvasive modality for the study of coronary arteries. The introduction of multislice computed tomography (MSCT) with retrospectively gated ECG reconstructions has substantially modified the coronary imaging scenario. In fact, the results have been promising since the beginning with 4-row MSCT. Currently the results reported with 16-row MSCT allow to question which role can the noninvasive coronary angiography play in clinical settings. With a scan of < 20 s after intravenous administration of iodinated contrast material it is possible to obtain information similar to conventional coronary angiography. Moreover, together with vascular patency, also atherosclerotic plaques are well visualized and characterized (type: calcified, soft, mixed). Ongoing studies are validating MSCT in the assessment of plaque vulnerability. The same technique allows to evaluate left ventricular function and myocardial wall motion impairment. Still with a few major limitations related to the heart rate, MSCT will broaden the noninvasive capability of evaluating coronary arteries in patients where conventional coronary angiography is exceeding the actual clinical question or considered as premature. PMID- 14558288 TI - [Acute embolic myocardial infarction complicating bioprosthetic stentless aortic valve endocarditis]. AB - We describe the case of bioprosthetic stentless aortic valve endocarditis due to Staphylococcus epidermidis in a female patient, who previously underwent mitral and aortic valve replacement: she presented left main coronary artery occlusion due to septic embolus with subsequent myocardial infarction complicated by cardiogenic shock. We discuss the therapeutic strategies, both medical and interventional, and the surgical options. PMID- 14558290 TI - [Atrioventricular nodal reentrant tachycardia with 2:1 anterograde block]. AB - We describe the case of a patient who presented with narrow QRS tachycardia at a rate of 100 b/min. The R-R intervals were constant, and negative P waves were evident in the inferior leads, midway between two ventricular complexes. A few minutes later, the tachycardia rate suddenly increased to 190 b/min, and the electrocardiographic pattern became typical of atrioventricular nodal reentrant tachycardia. Intravenous verapamil succeeded in restoring sinus rhythm. These data suggested an atrioventricular nodal reentrant tachycardia that at the beginning was associated with a 2:1 anterograde block. PMID- 14558289 TI - [Association of pulmonary embolism secondary to deep venous thrombosis and left atrial myxoma: case report and review of the literature]. AB - Left atrial myxoma is a benign tumor but it has a high prevalence of important complications. We report a case of a 73-year-old man affected by hypertension and chronic atrial fibrillation admitted to our hospital for pulmonary embolism, that resulted associated with a proximal deep venous thrombosis of the right leg and with an unknown left atrial myxoma. PMID- 14558291 TI - [Reply to "Open letter to Italian cardiologists. An invitation to debate" (Ital Heart J Suppl 2003, 4(4): 271-6)]. PMID- 14558292 TI - New evidence in support of the cardiovascular benefit of long-chain n-3 fatty acids. PMID- 14558293 TI - Electropharmacological effects of antiarrhythmic drugs on atrial fibrillation termination. Part I: Molecular and ionic fundamentals of antiarrhythmic drug actions. AB - In the last few years many studies have been performed with the aim of gaining a better understanding of the pathophysiological nature of atrial fibrillation. These recent observations provide new insights into the initiation and perpetuation of atrial fibrillation, underlying the importance of the pulmonary veins as major sources of atrial triggers and introducing new concepts such as the atrial electrical remodeling and the spatial heterogeneity of the electrophysiological characteristics of this arrhythmia. The increasing knowledge about the cardiac ion channel structure and function and about the electrophysiological actions of the antiarrhythmic drugs may contribute to a better comprehension of the mechanisms of the pharmacological termination of the arrhythmia. In part I of the review we try to give a unified vision of the old models and new concepts about the molecular and ionic fundamentals of antiarrhythmic drug actions. PMID- 14558294 TI - Electropharmacological effects of antiarrhythmic drugs on atrial fibrillation termination. Part II: New insights into the electrophysiological mechanisms of atrial fibrillation termination. AB - Antiarrhythmic drugs have been largely used to convert atrial fibrillation to sinus rhythm. Classes Ia, Ic and III antiarrhythmic agents are all known to be effective. Nevertheless, the electrophysiological properties of such agents even of the same class are very different. The mechanisms of the pharmacological termination of atrial fibrillation is an interesting issue that has not been so extensively studied yet. In this review we try to summarize the principal concepts about the electrophysiological substrate of atrial fibrillation and to give a unified and modern overview of the issue of the mechanisms of the pharmacological termination of the arrhythmia. PMID- 14558295 TI - Alterations of cardiac innervation in evolving myocardial infarction and transplanted hearts. An anatomo-clinical reappraisal. AB - BACKGROUND: Debate regarding the alterations of the cardiac innervation in an evolving myocardial infarction and transplanted hearts is still raging and most studies are based on radionuclide uptake of neurotransmitters or on the evaluation of the cardiorespiratory reflex. METHODS: The present investigation, upon human autoptic specimens of 57 infarcts and 8 cardiac transplants, was carried out with traditional neuropathology and modern molecular biology techniques. The specimens were selected for the identification of neurons, nerve fibers and their sheaths. RESULTS: First of all, these techniques confirmed the gross difference in the vulnerability of infarcted myocytes if compared with the local innervation, the metabolism of which is infinitely less oxygen-dependent than that of working myocardium (approximate quantitation below). Delicate technicalities of the traditional silver impregnation for nerves usually yield a large incidence of artifacts. Thereby, only perfect results (20% of cases), corroborated by parallel nerve sheath immunostaining (70% of cases), were retained and documented herein. In the meantime, acidosis and free radicals increase, while catabolite accumulation supervenes. These three factors threaten myocardial viability. Thereby, nervelets can be seen to survive the hyperacute phase of ischemia, but may be in part damaged by the successive granulocytic macrophage inflammation enzyme lysis of the infarcted muscle. The delayed and incomplete anatomical neural damage is confirmed by the observation of preserved nerve sheaths and neural filaments surviving in postinfarction scars, almost devoid of myocardiocytes. CONCLUSIONS: The rich sympatho-vagal cardiac network might further provide alternative bypasses for post-infarct reinnervation. The functional implications of this process remain unclear. PMID- 14558296 TI - Sixteen-year results of composite aortic root replacement for non-dissecting chronic aortic aneurysms. AB - BACKGROUND: The aim of this study was to evaluate the early and long-term outcomes in patients undergoing a Bentall procedure or its button modification for chronic aortic aneurysms with aortic valve incompetence. METHODS: Between January 1986 and January 2002, 65 patients (84% males, mean age 58.9 +/- 11 years) underwent aortic root replacement with a Bentall or a button-Bentall operation. Annuloaortic ectasia was the most frequent cause of aortic disease in this series (n = 37, 56.9%), followed by atherosclerotic aneurysms (n = 22, 33.9%), and post-stenotic dilation (n = 5, 7.7%). One patient (1.5%) underwent redo aortic root replacement,3 (4.6%) had a Marfan syndrome, and 6 (9.2%) underwent a concomitant replacement of the aortic arch. The duration of follow-up ranged from 2 to 192 months (mean 89.6 +/- 21.8 months). RESULTS: The 30-day mortality was 0%. Early non-fatal complications comprised: bleeding requiring surgical re-exploration (n = 1, 1.5%), low output syndrome (n = 1, 1.5%), acute renal insufficiency (n = 1, 1.5%), transient ischemic attack (n = 2, 3.1%), stroke (n = 1, 1.5%), and pulmonary insufficiency (n = 1, 1.5%). There was a late death due to a pulmonary neoplasm. The 16-year actuarial survival was 97 +/- 2% (hazard 0.02 +/- 0.02). No patient required reoperation. Furthermore, the long term clinical follow-up was characterized by the complete absence of endocarditis, anticoagulant-related hemorrhage, valve thrombosis, and prosthetic failure. Finally, the NYHA functional status was significantly improved (1.1 +/- 0.50, p < 0.001 vs preoperatively). CONCLUSIONS: The late results of the Bentall and button-Bentall procedures were excellent. Our findings confirm that these techniques still constitute the gold standard in the surgical treatment of combined valve and ascending aorta pathologies. PMID- 14558297 TI - Trends in cardiovascular risk factors and their treatment among patients discharged from divisions of internal medicine. The FAPOI-1 and FADOI-2 studies. AB - BACKGROUND: There is an increasing interest in monitoring cardiovascular risk factor levels and their treatment. Two studies were run in patients discharged from Divisions of Internal Medicine. METHODS: Two studies, respectively conducted in 1996 and 1999, have collected data on the diagnoses of cardiovascular diseases and cardiovascular risk factor levels and treatment in 6450 and 8133 patients (age > or = 35 years) discharged during 1 week from 235 and 345 Italian Divisions of Internal Medicine. RESULTS: The second survey revealed: a relative increase in heart failure and cerebrovascular disease diagnoses; lower serum levels of total cholesterol and triglycerides, lower systolic and diastolic blood pressure mean levels, and a decreased incidence of the habit of smoking among males. The proportion of treated hypertensives (blood pressure > or = 140/90 mmHg, or the use of antihypertensive drugs) increased slightly (65 to 68% among men and 69 to 71% among women). A reduction in the use of ACE-inhibitors was observed, balanced by an increase in the use of angiotensin II antagonists. The proportion of treated dyslipidemic patients (total cholesterol levels > or = 250 mg/dl, or HDL cholesterol levels < 35 mg/dl or triglyceride levels > or = 200 mg/dl or the use of hypolipidemic drugs) significantly increased from 10 to 25% among males and from 12 to 25% among females, due to a substantial increase in the use of statins. CONCLUSIONS: During a 3-year period increasing attention for cardiovascular risk factors and their treatment among patients discharged from Divisions of Internal Medicine has been documented. In particular, there has been a large increase in the use of statins, although large segments of patients may need further attention and drug treatment. PMID- 14558298 TI - Treatment of mediastinitis using an open irrigation and delayed sternal reconstruction with a pectoralis major muscle flap. AB - BACKGROUND: Mediastinitis is a very serious complication after cardiac surgery. To date, the optimal treatment of mediastinitis is still controversial: the "closed wound" procedures and the "open wound" treatments are the two conventional modalities reported in the literature. METHODS: Between January 1995 and December 2000, 20 patients, who had previously been submitted to cardiac surgery, were treated by a modification of the "open wound" treatment strategy for postoperative mediastinitis. All patients were scheduled for 2, 6, and 12 month clinical follow-up. The procedure performed consisted of three major steps: 1) early sternum reopening, followed by phase 2) including irrigation of the wound 3 times daily, and the final step 3) of delayed reconstructive surgery using the pectoralis major myocutaneous advancement flap closure technique. We prospectively analyzed the short- and long-term results of these procedures. RESULTS: The overall duration of hospitalization was 25 +/- 10 days; no patient required intensive care unit permanency. Clinical success was achieved in all 20 cases (100%). No recurrences of local (such as fistulas or abscesses) or systemic infections were noted, and no patient required sternal reopening during follow up. An optimal cosmetic result was obtained in all patients and only 2 cases had persistent sternal pain regressing at the 6-month follow-up control. CONCLUSIONS: Our data suggest that for patients with severe mediastinitis, this treatment strategy is safe. The clinical and esthetic success rates are high, the recovery time rapid, and the rates of short- and long-term complications very low. PMID- 14558299 TI - Nurses' observational study on the practice of secondary prevention in a cardiovascular department. AB - BACKGROUND: Although interventional studies have established the prognostic importance of the control of risk factors in patients with cardiovascular disease, reviews invariably show that the implementation of secondary prevention in patients with cardiovascular disease during hospitalization for acute manifestations or interventions is unsatisfactory. The aim of this study was to observe secondary prevention practice in a cardiovascular department, as part of a quality assurance program. METHODS: Two hundred and twenty patients discharged from the intensive coronary care unit, cardiac surgery unit and vascular surgery unit were prospectively included. Data were extracted from medical records and discharge documents. One hundred and eleven patients with at least one modifiable risk factor which was previously not corrected, were interviewed at discharge and were reassessed 3 months later. RESULTS: Written prescriptions about smoking cessation and weight reduction were given to 7 and 3% of smokers and overweight patients respectively. In 17% of patients no lipid measurement was reported, and in 49% of patients with low-density lipoprotein (LDL) cholesterol levels > 129 mg/dl statins were not prescribed. In patients with a history of infarction, aspirin and beta-blockers were prescribed in 90 and 64% respectively. In diabetics, statins were prescribed to 48% and angiotensin-converting enzyme inhibitors to 31%. Less than 40% of patients were able to refer appropriate levels for their blood pressure, weight, and cholesterol, and 30% fully comprehended the importance of smoking cessation. At the 3-month follow-up visit, 37% of patients had LDL cholesterol levels > 129 mg/dl--in half of these patients despite statins. In 61% of diabetics glycemic control was poor, and one third of smokers had not stopped smoking. CONCLUSIONS: These observations by the nurses have shown pitfalls in the implementation of guidelines, due to incomplete risk assessment, insufficient drug treatment and ineffective patient education. These data are the starting point for upcoming actions of quality improvement in the cardiovascular department of our hospital. PMID- 14558301 TI - Spontaneous closure of postinfarction ventricular septal rupture. A case report. AB - Spontaneous closure of a postinfarction ventricular septal rupture is extremely rare. We present such a case in which the postinfarction ventricular septal rupture closed spontaneously during follow-up. We postulate that the spontaneous closure of the ventricular septal rupture was probably due to thrombosis in the apical and septal aneurysm. PMID- 14558300 TI - Hypertrophy at ECG and its regression during treatment survey (HEART survey). Rationale, design and baseline characteristics of patients. AB - BACKGROUND: Left ventricular hypertrophy (LVH) detected at electrocardiography (ECG) is a predictor of an increased cardiovascular risk in essential hypertension. However, uncertainty remains concerning the reproducibility of ECG LVH and the prognostic relevance of its regression over time in hypertension. The aim of this study was to determine the prognostic value of baseline ECG LVH and its serial changes in a large cohort of hypertensive patients. METHODS: The Hypertrophy at ECG and its Regression during Treatment Survey (HEART Survey) is a prospective observational study conducted in 66 Italian centers. Inclusion criteria are essential hypertension with ECG LVH defined by the Perugia score (Cornell voltage criteria and/or a typical left ventricular "strain" pattern and/or a Romhilt-Estes score > or = 5 points) in subjects aged 45-84 years. The treatment of hypertension and other risk factors accords with current guidelines and is individually tailored. ECG is recorded twice at entry and periodically repeated over a 4-year follow-up period. Expert readers (unaware of the clinical findings) classify ECG. The incidence of major cardiovascular events in relation to baseline ECG and its changes over time are assessed, together with the reproducibility in the two baseline recordings. Overall, 708 patients aged 64 +/- 9 years have been enrolled in centers from northern (27%), central (32%) and southern (41%) Italy. Their baseline characteristics are presented. Follow-up is ongoing. CONCLUSIONS: The HEART Survey will examine the prognostic value of baseline ECG LVH and of its regression over time in a wide population of hypertensive patients. PMID- 14558302 TI - Transmural coronary inflammation triggers simultaneous multivessel rupture of unstable plaques. AB - The authors describe a case of sudden cardiac death caused by the simultaneous multivessel rupture of unstable atherosclerotic plaques, triggered by a transmural inflammatory process (coronaritis). Male subject, 44 years old, apparently in good health until 1 hour before death, when he complained of worsening dyspnea. At autopsy, it was found that the heart weighed 486 g. Evaluation of the coronary arteries revealed the presence of atherosclerotic plaques resulting in a lumen critical stenosis of the left anterior descending artery (LAD), right coronary artery (RCA) and left circumflex artery, and acute occlusive thrombosis of the LAD and RCA. Transverse sections of the ventricular mass highlighted the presence of eccentric hypertrophy of the left ventricle associated with myocardiosclerosis of the posterior interventricular septum and of the posterior wall of the left ventricle. Histology revealed the presence of a coagulative myocytolysis ascribable to the free walls of the left ventricle, and a focus of lymphocytic-active myocarditis. All coronary arteries were sites of intima fibroatheromatous plaques complicated by rupture and thrombosis within the RCA and LAD and by a transmural infiltrate consisting of macrophages and T lymphocytes associated with consensual medionecrosis and perineuritis. In conclusion, the present case report confirms the hypothesis that inflammation plays a key role in the onset of acute coronary syndromes as it promotes the formation of an unstable plaque as well as its rupture. PMID- 14558303 TI - Mid-ventricular paradoxical dynamic obstruction in a right-sided secondary hypertrophic cardiomyopathy. AB - Isolated right ventricular (RV) hypertrophy is a common diagnosis at echocardiography. Mid-ventricular obstructive involvement represents however a really unusual finding. Currently available studies on right-sided hypertrophic cardiomyopathy, regardless of whether they are associated with RV outflow tract obstruction, are usually limited to single cases. Besides, there is still no agreement about the clinical relevance and management of RV hypertrophy. In the present study the authors describe the echocardiographic findings of a patient presenting with RV hypertrophy in whom a rare coexistence of both fixed RV outflow tract and intraventricular dynamic obstruction was observed. Some functional effects of verapamil treatment are also discussed. PMID- 14558304 TI - One biologic marker (carbohydrate antigen-CA 125), two different diseases (ovarian cancer and congestive heart failure): practical implications of monitoring CA 125 serum levels. A case report. AB - Carbohydrate antigen (CA) 125 is a glycoprotein produced by the serous epithelium, found to be increased in ovarian cancer and currently used in the follow-up of patients with this malignancy and to evaluate the response to therapy. However, an increase in the blood levels of this tumor marker has been recently reported even in patients with congestive heart failure. We report a case of a woman in whom previously high levels of CA 125 were related to the presence of an ovarian cancer; after bilateral oophorectomy and chemotherapy, the CA 125 levels remained within the normal range for several years, until a new increase was recently detected, raising the suspicion of a cancer recurrence. Clinical evaluation, together with comprehensive laboratory assessment, allowed us to exclude a malignancy and to identify the presence of congestive heart failure due to left ventricular systolic dysfunction as being responsible for the abnormal CA 125 levels. The patient's clinical improvement following medical therapy was associated with a reduction in the serum levels of CA 125 to normal values. The clinical relevance of this finding is discussed. PMID- 14558306 TI - Perspectives. Part 1: Is hospital have/have-not gap worsening? PMID- 14558305 TI - Unusual echocardiographic aspect of the right coronary artery. PMID- 14558307 TI - National consensus conference on pertussis, Toronto, May 25-28, 2002.. PMID- 14558308 TI - Courage and commitment. PMID- 14558309 TI - A forum for graduate nursing students. PMID- 14558312 TI - Promoting healthy school communities. PMID- 14558313 TI - Developing regional clinical pathways in rural health. PMID- 14558314 TI - Reducing the risks in bariatric patient handling. PMID- 14558315 TI - International nurse applicants. PMID- 14558316 TI - Intermittent auscultation of the fetal heart rate. PMID- 14558317 TI - Teach what you know. Interview by Barbara Sibbald. PMID- 14558318 TI - Trends and determinants of contraceptive method choice in Kenya. AB - This study uses data from the 1989, 1993, and 1998 Kenya Demographic and Health Surveys to examine trends and determinants of contraceptive method choice in Kenya. The analysis, based on two-level multinomial regression models, shows that, over time, the use of modern contraceptive methods, especially long-term methods, is higher in urban than in rural areas, whereas the pattern is reversed for traditional methods. Use of barrier methods among unmarried women is steadily rising, but the levels remain disappointingly low, particularly in view of the HIV/AIDS epidemic in Kenya. One striking result from this analysis is the dramatic rise in the use of injectables. Of particular program relevance is the notably higher levels of use of injectables among rural women, women whose partners disapprove of family planning, uneducated women, and those less frequently exposed to family planning media messages, compared with their counterparts who have better access to services and greater exposure to family planning information. PMID- 14558319 TI - How gender roles influence sexual and reproductive health among South African adolescents. AB - Although the literature on Africa increasingly adopts a gendered approach to sexual and reproductive health issues, few studies have addressed adolescent pregnancy and parenthood in such a framework. This article examines links between gender ideology or gender roles and the social impact of adolescent childbearing in the lives of rural and urban adolescents in KwaZulu/Natal, South Africa. It employs a triangulated research methodology (focus-group discussions, narrative role playing and discussions, and questionnaires and in-depth interviews) to inform an analysis of adolescents' notions of male and female gender ideals. This analysis forms the basis for an exploration of the potential influence of adolescent childbearing on young peoples' lives and factors that shape their sexual and reproductive well-being. Results indicate that gender ideals are grounded in traits that reinforce poor sexual negotiation dynamics and behavioral double standards and that place adolescents at risk for early pregnancy and other sexual and reproductive health complications. Overall, adolescent parenthood is viewed negatively by participants of both sexes because it compromises personal, professional, and financial aspirations. Compared with its effect on boys, parenthood has a disproportionate (and highly negative) impact on girls that is directly linked to gender-based inequities. The article addresses the research and policy implications of these findings. PMID- 14558320 TI - Rural Indian women's care-seeking behavior and choice of provider for gynecological symptoms. AB - This study uses data from the India National Family and Health Survey-2 conducted in 1998-99 to investigate the level and correlates of care-seeking and choice of provider for gynecological symptoms among currently married women in rural India. Of the symptomatic women surveyed, 31 percent sought care, overwhelmingly from private providers (70 percent). Only 8 percent of women consulted frontline paramedical health workers. Care-seeking behavior and type of providers consulted varied significantly across different Indian states. Significant differentials in care-seeking by age, caste, religion, education, household wealth, and women's autonomy suggest the existence of multiple cultural, economic, and demand-side barriers to care-seeking. Although socially disadvantaged women were less likely than better-off women to consult private providers, the majority of even the poorest, uneducated, and lower-caste women consulted private providers. Geographical access to public health facilities had no significant association with choice of provider, whereas access to private providers had only a moderately significant association with that choice. The predominance of use of private services for self-perceived gynecological morbidity warrants the inclusion of private providers in the national reproductive health strategy to enhance its effectiveness. PMID- 14558322 TI - Inconsistent reporting of female genital cutting status in northern Ghana: explanatory factors and analytical consequences. AB - Although many cross-sectional social surveys have included questions about female genital cutting status and correlated personal characteristics, no longitudinal studies have been launched that permit investigation of response biases associated with such surveys. This study draws upon the findings of a longitudinal study of women aged 15 to 49 in rural northern Ghana. The self reported circumcision status of women interviewed in 1995 was compared with the status they reported when they were interviewed again in 2000 after the government began enforcing a law banning the practice and public information campaigns against it were launched. In all, 13 percent of respondents who reported in 1995 that they had been circumcised stated that they had not been circumcised in the 2000 reinterview; this inconsistency reached 50 percent for the youngest age group. Analysis shows that women who said they had not been circumcised are significantly younger, more likely to be educated, and less likely to practice traditional religion than are women who reported that they were circumcised. Factors that may explain these correlates of denial are discussed, and implications for research are reviewed. PMID- 14558321 TI - Factors influencing young Malians' reluctance to use hormonal contraceptives. AB - During a qualitative evaluation of three peer-education programs in urban Mali, young people stated that they were wary of using either the pill or injectable contraceptives because they believed that these methods would make them sterile. Unmarried women's contraceptive decisionmaking was not primarily driven by a current need to limit fertility, but rather by a future need to maximize it in order to gain status through childbearing in their marital households. Further interviews explored notions of conception, menstruation, and the perceived action of hormonal methods on the reproductive system. Findings revealed that menstrual disruption (in the form of amenorrhea or prolonged bleeding) appeared to have dire repercussions, including accusations of witchcraft and immoral behavior that could result in a woman's being divorced or in her husband's acquiring an additional wife. The social consequences of side effects were perceived to be more important than their biological manifestations, and together with the fear of sterility, resulted in a preference for the condom. PMID- 14558323 TI - Colombia 2000: results from the Demographic and Health Survey. PMID- 14558324 TI - Haiti 2000: results from the Demographic and Health Survey. PMID- 14558326 TI - [Daily hemodialysis]. PMID- 14558325 TI - [Therapeutic approaches of nephrotic syndrome in children at the beginning of the XXI century]. PMID- 14558327 TI - [Pure red cell aplasia and antibodies against erythropoietin. Doubts still remain]. PMID- 14558328 TI - [Fungal peritonitis in peritoneal dialysis: the new approaches can become a hope]. PMID- 14558329 TI - [Spanish survey on anaemia management]. AB - The European Best Practice Guidelines for the Anaemia Management (EBPG) represents the main reference for the treatment of patients with chronic renal failure (CRF). This study was developed two years after EBPG's publication in order to evaluate the impact of theses guidelines in the clinical practice. This is an epidemiological, multicentric and prospective study including patients on dialysis and erythropoietin treatment. The aetiology of CRF, prevalence and anaemia treatment, concurrent pathologies and side effects had been evaluated during follow-up. The inclusion period was june-august 2000. Patients were evaluated during six months. Two thousands six hundred and eighteen dialysis patients were recruited from different national centres (2,171 on haemodialysis and 447 on peritoneal dialysis). Fifty one percent of haemodialysis patients and forty one percent of patients on peritoneal dialysis started erythropoietin treatment with a basal haemoglobin > 9 g/dL (mean 8.9 g/dL and 9.2 g/dL, respectively). At baseline, 35% of patients had haemoglobin < 11 g/dL (mean 11.4 g/dL), but at the study end, 21.2% had haemoglobin < 11 g/dL (mean 11.8 g/dL). Seventy nine percent of patients were treated with erythropoietin using subcutaneous route: 27.5% with once a week regimen, 33.0% in two weekly doses and 39.5% in three weekly administrations. No differences in doses requirements were showed by administration route. These results seem to support that there already exists a good knowledge of the International Guidelines for the anaemia management, but there is still a gap between them and the results in the clinical setting. In general erythropoietin treatment starts late and at low haemoglobin levels. However, close monitoring of haemoglobin levels seems to improve patient's outcomes. PMID- 14558330 TI - [Role of transforming growth factor beta-1 gene polymorphisms in the development of chronic allograft nephropathy in renal transplant recipients]. AB - BACKGROUND: Chronic allograft nephropathy (CAN) is the main cause of graft loss after the first year of transplantation, and renal biopsies show a predominance of fibrotic lesions. Human transforming growth factor beta-1 (TGF-beta 1) is the principal profibrogenetic cytokine which has been recently implicated in the development of CAN. Seven TGF-beta 1 gene polymorphisms have been recently described and some of them have been related to the development of several diseases. AIM: To analyse the relationship between TGF-beta 1 gene polymorphisms and the development of CAN in a group of renal transplant patients with a long term follow-up. METHODS: A restriction enzyme-based method for TGF-beta 1 genotyping was used to detect four TGF-beta 1 gene polymorphisms in codon 10, 25 and 5'-flanking region (-800 and -509 positions). A retrospective case-control study were performed on sixty renal transplant recipients with 8 years of post transplant, 22 of them with CAN (cases) and 38 with normal graft function (controls). We studied 73 subjects to analyse the distribution of the genotypes in the area. RESULTS: The genotype frequencies were similar in the study and control group. The presence of chronic allograft nephropathy was statistically associated with the combination Pro/Pro10 TT509 polymorphism in the TGF-beta 1 gene, and these patients develop chronic rejection more quickly than the rest of the patients. Chronic allograft nephropathy was also associated with delta age recipient-donor, with older donors being a significant risk factor for chronic nephropathy. The logistic regression analysis confirmed the independent role of TT509 Pro/Pro10 TGF-beta 1 polymorphism with a Odd Ratio of 5.8 (1.14-29.7) in chronic nephropathy being the delta age recipient-donor a confounder factor but not an effect modifier. The rest of the TGF-beta 1 gene polymorphisms and the classic risk factors were not associated with the development of chronic allograft nephropathy. CONCLUSIONS: These data suggest a leading role for TGF beta 1 gene polymorphisms (TT509 Pro/Pro10) in the development of chronic allograft nephropathy. The identification of this genetic predisposition to chronic allograft rejection could play a decisive role in the prevention of this common pathology. PMID- 14558331 TI - [Secondary amyloidosis (AA) and renal disease]. AB - Amyloidosis is a disease resulting from extracellular deposition of fibrillar protein in various organs. AA amyloidosis may complicate chronic inflammatory diseases, chronic infections and another chronic diseases. We review 31 patients (13 males and 18 females) with biopsy proven renal or rectal AA amyloidosis, referred to out hospital between january 1999 and november 2002. Renal failure was defined as serum creatinine > or = 1.5 mg/dl. Mean age was 58.4 +/- 15.7 years. The causes of AA amyloidosis were an underlying chronic rheumatologic disease (51.6%), chronic infection (41.9%) and a chronic inflammatory intestinal disorder (6.5%). Renal failure (RF) was detected in 20 patients (61.2%) and proteinuria and hematuria were found in 90.3% and 45.5 respectively. Proteinuria at diagnosis was 5.2 +/- 3.9 g/24 h and mean serum creatinine 3.5 +/- 3.7 mg/dl. Survival of patients without dialysis was 66.8 (51.1% RF, 90.9% non-RF) and 53.4% (38.2 RF, 77.9% non-RF) at 12 and 24 months respectively (p = 0.017). End-stage renal disease developed in 13 patients (41.9%). Ten patients were maintained on hemodialysis and 3 on CAD. Survival in dialysis at 6 and 12 months was 68.3% and 42.7% respectively. Fifteen patients died and the main causes of death were: infections (46.6) haemorrhagic complications (33.3%), cardiovascular events (13.3%) and cachexia (6.6%). PMID- 14558332 TI - [Whole PTH and 1-84/84 PTH ratio for the non invasive determination of low bone turnover in renal osteodysthrophy]. AB - The conventional intact PTH assays detect not only PTH 1-84 but also inactive fragments (as PTH 7-84) that accumulate in renal failure. There has been a recent development of a new PTH assay that measures only true 1-84 PTH (Whole PTH or CAP assay, Scantibodies). As 7-84 PTH fragment is antagonistic on bone effects of 1 84 PTH, Moniere-Faugere has suggested that 1-84/7-84 PTH ratio less than 1 is predictive of low turnover. We evaluated the usefulness of CAP assay and the 1 84/7-84 PTH ratio as markers of bone turnover in a groups of 24 patients in peritoneal dialysis (PD). Patients were classified as having low bone turn over if they had a Total PTH (similar to intact PTH) of less than 100 pg/ml. We also measured serum CrossLaps (CTX) as another serum resorption marker. Patients had a mean Whole PTH of 95.5 pg/ml and a mesan total PTH of 155.4 pg/l (range 9 to 900). Whole PTH represented 69.1% of total PTH. Fifteen patients (62.5%) had a total PTH of less than 100. These patients had a 1-84/7-84 relationship of 1.9 +/ 1.8 while 9 patients with Total PTH more than 100 had a relationship of 1.29 +/- 0.6 (p = NS). There was a tight correlation between Whole PTH and total PTH (r = 0.98; p < 0.0001) and with serum CTX (r = 0.78; p < 0.0001). We conclude that 1 84/7-84 ratio does not seem useful in the prediction of low bone turnover and that Whole PTH does not seem to be more useful than intact PTH in the prediction of bone turnover in this population. Future studies should correlate this markers with direct measurements of bone turnover in bone biopsies to demonstrate their usefulness in the prediction of the type of renal osteodystrophy. PMID- 14558333 TI - [An outbreak of gram-negative bacteremia (GNB), especially enterobacter cloacae, in patients with long-term tunnelled haemodialysis catheters]. AB - Vascular access through a venous catheter for haemodialysis is associated with increased risk of thrombosis, central venous stenosis, short access survival and inadequate dialysis. The most important catheter-related complications, which determine method survival, are infection and dysfunction. In particular, infectious episodes are in some studies the leading cause for untimely catheter removal and for catheter-related morbidity but also for morbidity in dialysis patients. Double-lumen central venous catheters used for haemodialysis, are common causes of septicaemia. Most cases are caused by staphylococci. Episodes of gram-negative bacteriemia have been traced to bacterial contamination of water and/or dialysate, errors in dialyzer reprocessing, and improper setup procedures. In this paper, we describe and outbreak of gram-negative bacteremia, firstly E. cloacae, in an outpatients haemodialysis unit, in the patients with long-term tunnelled haemodialysis catheters. We describe the epidemic investigation that we achieved to identify the source of contaminating bacteria and the route by which bacteria gained access to the bloodstream. We prove the contamination by gram negative bacterium of the water-distribution lines and haemodialysis machines. Moreover, E. cloacae strains isolated from the lines and machines are genotypically identical to the isolated from the patients. Also, we prove that the hands of health care personnel are unintentional carriers. The outbreak was finished when decontamination of dialysis machines was enhanced and dialyzer priming fluid was modified. PMID- 14558335 TI - [Acute renal failure secondary to hepatic veno-occlusive disease in a bone marrow transplant patient]. AB - Acute renal failure following bone marrow transplantation is a frequent complication with an incidence ranging 15-30% and with high rates of morbidity and mortality. Numerous potential etiologies can be implicated as chemotherapy regimen, use of nephrotoxic antibiotics, sepsis-induced damage, cyclosporine toxicity and other especific pathologies as graft-v-host disease or veno occlusive disease of the liver. We report the case of a 41-year-old man who underwent autologous peripheral blood stem cell transplantation and developed and acute renal failure secondary to a fatal veno-occlusive disease of the liver. Incidence, potential predisposing factors, outcome and possibilities of treatment are reviewed. PMID- 14558334 TI - [Evaluation of methods to calculate dialysis dose in daily hemodialysis]. AB - Daily dialysis has shown excellent clinical results because a higher frequency of dialysis is more physiological. Different methods have been described to calculate dialysis dose which take into consideration change in frequency. The aim of this study was to calculate all dialysis dose possibilities and evaluate the better and practical options. Eight patients, 6 males and 2 females, on standard 4 to 5 hours thrice weekly on-line hemodiafiltration (S-OL-HDF) were switched to daily on-line hemodiafiltration (D-OL-HDF) 2 to 2.5 hours six times per week. Dialysis parameters were identical during both periods and only frequency and dialysis time of each session were changed. Time average concentration (TAC), time average deviation (TAD), normalized protein catabolic rate (nPCR), Kt/V, equilibrated Kt/V (eKt/V), equivalent renal urea clearance (EKR), standard Kt/V (stdKt/V), urea reduction ratio (URR), hemodialysis product and time off dialysis were measured. Daily on-line hemodiafiltration was well accepted and tolerated. Patients maintained the same TAC although TAD decreased from 9.7 +/- 2 in baseline to a 6.2 +/- 2 mg/dl after six months, p < 0.01. No significant changes were observed in weekly Kt/V and eKt/V throughout the study. However EKR, stdKt/V and weekly URR were increased during D-OL-HDF in 24-34%, 46% and 50%, respectively. Hemodialysis product was raised in a 95% and time off dialysis was reduced to half. CONCLUSION: Dialysis frequency is an important urea kinetic parameter which there are to take in consideration. It's necessary to use EKR, stdKt/V or weekly URR to calculate dialysis dose for an adequate comparison between different frequency dialysis schedules. PMID- 14558336 TI - [Acute interstitial nephritis induced by loratadine]. AB - Loratadine is a second generation histamine H1 receptor antagonist, that has high potency antiallergic properties and is associated with low adverse effects compared with other antihistamines. Acute interstitial nephritis is a cause of acute renal failure that is most often induced by drugs or, less frequently, infection or sarcoidosis. Although the number of drugs associated with acute intersticial nephritis is too large, the antihistaminic loratadine have never been reported before. We report a case of an interstitial nephritis with acute renal failure that suggesting hypersensitivity reaction in a 77 old man who had received loratadine (10 mg/day) during ten days before his assessment to our hospital by disseminated pruritic syndrome. The initial suspect was rapidly progressive glomerulonephitis and renal biopsy was practice and treatment with corticosteroids were initiated (prednisone bolus of 500 mg three days and 1 mg/kg/day/later). The loratadine therapy was cessation. He exhibiting a slow and progressive improvement on renal function and one month later, urea and creatinine levels was normal and hematuria and proteinuria had disappeared. The corticosteroids therapy were progressive decreased until withdrawal. We think that this is an interesting case, basing in its clinical presentation and that it had never been reported before. PMID- 14558337 TI - [Vasculitis in elderly patients associated to ANCA]. PMID- 14558338 TI - [Coronary artery calcification in a patient in hemodialysis]. PMID- 14558339 TI - [Is it cost-benefit acceptable the change of EPO subcutaneous to intravenous administration in patients in hemodialysis?]. PMID- 14558340 TI - [Treatment of multiresistant infections : filgastrim, an alternative for the treatment of vascular accesses infections in hemodialysis]. PMID- 14558341 TI - [Model corasol-induced seizures are followed by increase of nitric oxide generation and are abolished by mexidol and alpha-tocopherol]. AB - The effect of mexidol and alpha-tocopherol on the onset and development of acute epilepsy model was studied in Wistar rats using penthylenetetrazole induced convulsions. The intensity of the nitric oxide (NO) production in the cerebral cortex was determined by a direct method using electron paramagnetic resonance. The rate of lipid peroxidation (LPO) was estimated by measuring the level of secondary products (thiobarbituric acid reactive species). The peak of penthylene tetrazole convulsions is accompanied by a significant increase in the levels of both NO and LPO products. Mexidol (150 mg/kg) and alpha-tocopherol (100 mg/kg) hindered the development of model convulsions, prevented NO buildup, and inhibited LPO growth. It is suggested that suppression of the excess NO production in the cortex and inhibition of the LPO enhancement can be involved in the mechanism of action of antiepileptic drugs. PMID- 14558342 TI - [Effect of pyracetam on the color discriminative function of retina in patients with craniocerebral trauma]. AB - The chronic administration of piracetam over a period of four weeks in patients after heavy craniocerebral traumas significantly improves the color discrimination of retina with respect to all four colors studied. It is suggested that the improved functioning is related to the nootrope effect upon the GABAergic processes both in the retina and in the related cerebral structures. PMID- 14558343 TI - [Effect of mexidol on the course of post-traumatic epilepsy]. AB - The results of a complex 10-day therapy of patients with posttraumatic epilepsy, involving the antioxidant mexidol, are presented. The efficacy of the therapy was assessed based on the data of clinico-neurological, instrumental, and immunological examinations before and after the treatment and evaluated in terms of the dynamics of the neurological state of the patients, EEG features, and immunological data (including the content of proinflammatory IL-1 beta cytokine and antiinflammatory IL-4 cytokine in the blood serum). Based on a comparison of the results obtained in patients receiving traditional antiepileptic drugs with and without mexidol, it is possible to recommend the complex treatment involving mexidol for the therapy of patients suffering of the postraumatic epilepsy. Mexidol provides for an additional immunocorrection and enhances the effect of antiepileptic drugs. PMID- 14558344 TI - [Effect of buspirone on aggressive and anxiety behavior of male mice with various aggressive experience]. AB - The effect of the 5-HT1A receptor agonist buspirone on the aggressive and anxious behavior of C57BL/6J male mice with different experience of aggression was studied. In the group of animals with short-time (3 days) winner experience, buspirone (1 mg/kg, i.p.) produced an anxiogenic effect (manifested in the plus maze test) and reduced the level of aggression (agonist confrontation test). No such effects were observed in the group of animals with a long-time (20 days) aggression experience. It is suggested that the antiaggressive effect of buspirone in the former case is related to the anxiogenic action, while a long experience of aggression reduced the pharmacological sensitivity of 5-HT1A receptors to buspirone in the latter case. PMID- 14558346 TI - [Cardiac effects of fenibut in development of experimental chronic renal insufficiency]. AB - The effect of fenibut on the mechanical activity of myocardium was studied in vitro and in vivo in rats with experimental chronic renal insufficiency (CRI) in a regime of physiologically alternating load simulating the intact heart function. The administration of fenibut (10 mg/kg) in rats after nephrectomy prevents the development of myocardial hyperfunction (characteristic of the animals with CRI in stage 1). In in vitro experiments on isolated myocardium fenibut also decreased the myocardial hyperfunction and reduced contractility to a control level, which was accompanied by accelerated relaxation in all finite systolic lengths. PMID- 14558345 TI - [Anti-ischemic properties of the novel antioxidant enoxifol]. AB - The antiischemic properties of the new antioxidant drug enoxifol, representing a derivative of N9-imidazo[1,2-a]benzimidazole, was studied in rats on the models of a double-vessel occlusion of common carotid arteries under normal conditions and on the background of irradiation. In addition, we studied the effect of enoxifol on the rehabilitation of experimental animals in the early reperfusion period after total brain ischemia. PMID- 14558347 TI - [Studying cGMP-dependent mechanisms of vinpocetine effect on smooth muscle cells]. AB - The results of the membrane potential measurements (by the double sucrose gap junction technique) and the smooth muscle tension determination (by the mechanical force measurements) in the rat aorta showed that vinpocetine potentiates the effect of sodium nitroprusside and nitroglycerin on the smooth muscle cells. In the concentration range of 2-20 microM, vinpocetine produced a dose-dependent inhibition of the Ca2+ conductivity of the membrane and decreased the smooth muscle contractility response. At a concentration of 1 microM, the drug acted as an inhibitor of the phosphodiestherase (PDE) activity and produced the effects similar top those of dibutyryl-cGMP (rather than dibutyryl-cAMP). In the presence of 10 microM of Methylene Blue (an inhibitor of the soluble fraction of guanylate cyclase), the cGMP-dependent effects of vinpocetine were suppressed on the background of 100 microM of sodium nitroprusside, but retained on the background of 10 microM of 3-isobutyl-1-methylxanthine (IBMX), a nonspecific PDE inhibitor. IBMX acted like dibityryl-cGMP and activated the K+ conductivity of the membrane. It is suggested that cGMP-dependent effects of vinpocetine are related to its action upon the Ca2+ and Na+ and (but not K+) conductivity and to the cGMP-induced increase in the contribution of sarcoplasmic calcium to the contractile response. PMID- 14558348 TI - [Comparative choleretic properties of natural xanthone compounds from Gentianopsis barbata]. AB - Comparative data are presented for the choleretic activity of xanthones isolated from Gentianopsis barbata (Froel.) Ma. (Gentianaceae family): a sum of xanthone glycosides (I) and aglycons (II); xanthone aglycons, including decussatin [1-(OH) 3,7,8-(OCH3)3] (III) and gentiacaulein [1,7-(OH)-3,8-(OCH3)2] (IV); and xanthone glycoside gentiabavaroside [1-O-primverosyl-7-(OH)-3,8-(OCH3)2] (V). It was established that (II) is superior to (I) with respect to cholagogic effect and is inferior to (III) and (IV) with respect to cholatostimulant action. PMID- 14558349 TI - [Experimental systemic enzyme therapy of gouty and primary glomerulonephritis]. AB - The influence of a systemic enzymotherapy on the morphological, biochemical, and functional manifestations of the kidney damage during the experimental gouty and primary glomerulonephritis is described in comparison to the results obtained by traditional methods. PMID- 14558350 TI - [Experimental progestogenic activity of novel derivatives of 17alpha hydroxyprogesterone]. AB - Experiments on infantile female rabbits showed that new 17a-hydroxyprogesterone derivatives (AMOL phenyl propionate, AMOL isopropyl ester) possess a pronounced gestagenic activity. In particular, AMOL isopropyl ester is capable of maintaining pregnancy in ovariectomized animals (Corner-Allen assay). PMID- 14558351 TI - [Effect of phensuccinal on lipid metabolism, lipid peroxidation, and antioxidant system activity in rabbits with dithiazone-induced diabetes]. AB - A three-month administration of phensuccinal improved glucose homeostasis, decreased the levels of total cholesterol, triglycerides, fatty acids, and low density lipoproteins in the blood serum, and reduced the lipid peroxidation rate as compared to the untreated diabetic control. In addition, phensuccinal increased the content of the antiatherogenic high-density lipoprotein fraction and the related paraoxonase enzyme activity. The preventive effect of phensuccinal with respect to diabetic dyslipidemia development, together with the antioxidant action, show this compound to be a promising therapeutic means of preventing and/or reducing macrovascular complications in diabetic patients. PMID- 14558353 TI - [Efficacy of enzyme-inducing agents in experimental tetrachloromethane poisoning]. AB - Liver monooxygenase system inductors benzonal, galonal, and galodif exceed phenobarbital and zixorin in therapeutic efficacy with respect to rats poisoned with tetrachloromethane. The former drugs increase the content and catalytic activity of cytochrome P-450, improve conjugation with reduced glutathione, prevent the cytolysis of hepatocytes, and stimulate the bile secretion and the liver excretion function. PMID- 14558352 TI - [Preclinical prognosis of pyracetam and picamilon safety based on acute toxicity data]. AB - A comparative acute toxicity test of the nootropic drugs piracetam and picamilon was performed on rats. The study was based on the principles of integral evaluation of the drug effect upon the functional and behavioral state of animals. It was found that the conventional therapeutic index does not coincide with the actual therapeutic activity range. Piracetam and picamilon, while exhibiting significantly different toxicity, are characterized by approximately equal ranges of the therapeutic activity. PMID- 14558354 TI - [Comparative evaluation of effects of p-tyrosol and Rhodiola rosea extract on bone marrow cells in vivo]. AB - The effects of p-tyrosol and Rhodiola rosea extract on the hemopoietic system were compared on a model of subacute lead intoxication. No significant differences between the activity of two preparations were revealed by the study of plasma membrane blebbing, apoptosis, and necrosis processes in bone marrow. At the same time, p-tyrosol exhibited a more pronounced effect upon lipid peroxidation and offered significant protection against lead intoxication. PMID- 14558355 TI - [Succivil efficacy in endogenous intoxication]. AB - Succivil, a drug based on the blood substitute succinasol and containing 1,4 naphthoquinone, was studied as a means of restoring the living activity of cells occurring under extremal conditions of alcohol, heliotrin, or combined intoxication. A thorough investigation of the role of the free-radical oxidation in the dynamics of cell damage, in combination with histological, morphological, and biochemical data, showed that the aforementioned extremal intoxication factors lead to disorders in the cell energetics and intracell detoxicant system and to more or less manifested changes in the hemodynamic, biochemical, and physiological parameters. Succivil is capable of reanimating the cells damaged by the extremal factors studied. PMID- 14558356 TI - [Study of pharmacological properties of betulin bishemiphthalate]. AB - The results of pharmacological tests showed that betulin bishemiphthalate possesses hepatoprotector, antioxidant, and immunotropic properties. Administered in combination with hydroxymethyluracil, that betulin bishemiphthalate prevented the loss of experimental animals upon irradiation. PMID- 14558357 TI - [Effect of natural enterosorbents in experimental liver disorders]. AB - The effect of enterosorbents of natural origin, zoosterol and sodium alginate, on the main types of metabolism and the macro- and microelement composition (calcium, magnesium, zinc, copper) of the renal tissues was studied in animals with experimental nephrocalcinosis. Increased complex-forming ability of zoosterol, related to a higher absorption activity of low-molecular-weight fragments of this enterosorbent, favors retention of large molecules in the form of calcium salts in the parenchyma. Sodium alginate favors the elimination of calcium compounds and compensates for calcification of the renal tissue, which allows this this enterosorbent to be used as a crystal-inhibiting complex-forming agent for correcting metabolic dysbalance in cases of nephrocalcinosis. PMID- 14558358 TI - [Mechanisms of antistressor activity of Inula helenium preparations]. AB - The antistressor and antioxidant properties of preparations from the underground part of elfwort (Inula helenium) were studied on a model of acute stress in mice. The preparations produced a protective effect with respect to internal organs, blood, carbohydrate metabolism, and lipid peroxidation process. PMID- 14558359 TI - [Correlation between chemical structure and a target as basis for classification of direct-acting antioxidants]. PMID- 14558360 TI - [New aspects of estrogenic effect]. PMID- 14558361 TI - ROI and IT: strategic alignment and selection objectivity. PMID- 14558362 TI - Return on investment: going beyond traditional analysis. PMID- 14558363 TI - In search of the ROI from CPOE. PMID- 14558364 TI - Creating a healthy relationship with your (next) IT vendor. PMID- 14558366 TI - Life after go-live. Part 4: Preventing error with an EMR. PMID- 14558365 TI - At least 44,000 patient deaths this year: how do you measure ROI? PMID- 14558367 TI - Pam Arlotto, healthcare IT strategist, shares insight on the ROI Process and IT. Interview by Richard D Lang. PMID- 14558368 TI - Finding value from IT investments: exploring the elusive ROI in healthcare. AB - This article explores the historical IT value research, discusses its applicability to IT investments in healthcare, and highlights how it is challenged by several factors unique to the healthcare industry. The integration of historical IT value research with healthcare industry attributes provides an important context for understanding why the IT value proposition in healthcare has been so elusive. The article also poses a set of guidelines, which, based on the IT value research outside of healthcare, may assist in alleviating some of the current frustration with determining the value of healthcare IT investments. PMID- 14558369 TI - Who's counting now? ROI for patient safety IT initiatives. AB - The impact and expectation of cost-justifying patient safety IT initiatives using a traditional ROI must evolve to focus beyond the financial benefit. It must encompass overall patient safety, patient satisfaction, and employee and physician satisfaction benefit categories. Computerized physician order entry (CPOE) and bar code medication administration (BCMA) systems are two particular clinical point-of-care products that will play a key role in addressing patient safety objectives. Integrating the two technologies can bring both financial and clinical benefits. PMID- 14558370 TI - Clinical ROI: not just costs versus benefits. AB - Although sophisticated economic modeling can be used to quantify intangible benefits, ROI calculations for clinical information systems are driven more by the values and strategic direction of an organization than by any other considerations. But investing in clinical information tools to ensure quality and patient safety is, in reality, required as a cost of doing business and functioning as a safe hospital. PMID- 14558371 TI - The New England Healthcare EDI Network. AB - The New England Healthcare EDI Network (NEHEN) is a collaborative of providers and payers in eastern Massachusetts that created, manages, and operates a shared insurance EDI infrastructure. NEHEN currently has 12 provider and three payer members, and supports over 1,000,000 insurance EDI transactions per month. This paper describes the philosophies that define the NEHEN business model and discusses its governance structure, technology, operational issues associated with its implementation, and its current status, along with lessons learned from the NEHEN undertaking. PMID- 14558372 TI - Analyzing computer based patient records: a review of literature. AB - A wide-ranging literature review of computer-based patient record (CPR) implementation over the past decade reveals that clinical, workflow, administrative, and revenue enhancement benefits of the CPR outweigh barriers and challenges--but only if healthcare organizations redesign certain work processes. Among other key efforts, organizations must train and motivate users to navigate CPR systems, as well as develop a common structured language. Clinicians who used CPRs found that electronic access to clinical information saves time and provides a thorough and efficient way to manage patient information. PMID- 14558373 TI - Advancing the state of data integration in healthcare. AB - There is growing consensus that clinical information systems will provide the bridge to advancing the integration of information systems in healthcare. In spite of developments in technology that have enabled some organizations to integrate clinical information with care delivery in ways that can promote safer, more efficient patient care, the majority of healthcare has yet to achieve this goal. Why aren't we there yet? PMID- 14558374 TI - Health integration cost cutting. AB - By combining several traditional approaches to integration with newer innovations, healthcare organizations are finding that problems such as HIPAA remediation and providing secure online access to patient records can be resolved by leveraging integration projects across the enterprise. In the current cost cutting environment, the projects that get done are those that can be justified by direct impact on additional revenue, can show significant cost savings and extremely fast return on investment, and meet regulatory requirements. PMID- 14558375 TI - Antecedents to the adoption of ASPS in healthcare. AB - The objective of this exploratory study was to identify drivers of adoption for a new form of information technology outsourcing--the ASP model--in the healthcare industry. Primary data were collected in January 2002 from a nationwide survey of senior-level healthcare information technology executives. Cost management (supplier presence, asset specificity, production costs, transaction costs, resource availability) and relative advantage (reliability, customizability, strategic alignment, and magnitude of potential loss) were found to have the largest influences on adoption behavior. PMID- 14558376 TI - Electronic prescribing: ready for prime time? AB - Escalating drug expenditures and the prevalence of medication errors have prompted calls for drug cost control and quality improvement. E-prescribing promises to address these concerns. This study aims to describe the benefits and costs of eRx as assessed by key industry stakeholders, identify the components of a needed electronic prescribing infrastructure and the barriers to its completion, and identify potential public and private policies and initiatives that could encourage the development of an EPI and hasten eRx adoption among stakeholders. PMID- 14558377 TI - [Success rate of direct pulp capping with calcium hydroxide]. AB - Aim of this retrospective study was to evaluate the success rate of direct pulp capping treatment using calcium hydroxide-containing agents according to the age of the patient, the type of tooth and the person who performed the treatment. Over a period of seven years a total of 659 patients were treated. 763 direct pulp-cappings were recorded. From these 308 patients a total of 359 pulp-cappings (= 100%) were reexamined one to eight years after treatment. 45.1% of the reexamined teeth were in males and 54.9% in females. 61.7% of the treated teeth were in the upper and 38.8% in the lower jaw. Out of the 359 teeth (= 100%) the success rate was 61% (219 teeth); while 113 teeth (31.4%) had received a root canal treatment and 27 (7.6%) had been extracted. In this study the success rate depended on the group of teeth. It decreased continuously from the anterior teeth to the molars. The central incisors showed the highest (83.3%) and the wisdom teeth the lowest (38.9%) success rate. In addition, the success rate decreased with increasing age of the patients. The most successful treatments (76.4%) were registered in the group of patients, aged 10-19 years. PMID- 14558378 TI - [Ribotyping Fusobacterium nucleatum isolates from healthy and diseased periodontium]. AB - Fusobacterium nucleatum is one of the most frequently cultured bacteria from subgingival plaque. It can be isolated from active periodontal lesions as well as from healthy sites. Currently, F. nucleatum has been divided into five subspecies. An early hypothesis suggested an association between certain subspecies and periodontal disease or health. On the other hand, a broad heterogeneity between F. nucleatum isolates has been suggested, so that the subspeciation scheme and therefore the proposed association may not be valid. The purpose of the present study was to analyze if clonal differences exist between human F. nucleatum isolates from either periodontally healthy or diseased individuals. 23 F. nucleatum isolates from periodontally healthy and 17 isolates from periodontally diseased individuals were analysed by ribotyping. A broad genetic heterogeneity was present, although some of the isolates from periodontally healthy individuals were genotypically identical. However, no clonal differences between isolates from periodontally healthy or diseased individuals was detected. PMID- 14558379 TI - [Relationship between occlusion and myoarthropathy. Introduction of an integrating neurobiological model]. AB - For many decades, the relationship between occlusal factors and temporomandibular disorders has been discussed controversially in the dental literature. After a brief description of current points of view about this topic, a recently published clinical trial by Le Bell et al. (2002) is summarized. The results of this study are discussed on the basis of a novel neurobiologic model, which is based on the heterogeneous activation of the masticatory musculature. This model appears to be suitable to explain the pathogenic and therapeutic effects due to occlusal management of patients with myofascial facial pain. PMID- 14558380 TI - [Post-exposure prophylaxis against HIV]. PMID- 14558382 TI - [Clinical use of bone graft substitutes in orthopedic surgery. Status and future possibilities]. PMID- 14558381 TI - [Post-exposure prophylaxis after sexual or occupational exposure to HIV]. PMID- 14558383 TI - [Pregnancy and heart disease]. PMID- 14558384 TI - [Anakinra in the treatment of rheumatoid arthritis]. PMID- 14558385 TI - [The impact of repeated measurement of generic quality indicators]. PMID- 14558386 TI - [Practice variation in diagnosis and treatment of men with lower urinary tract symptoms by general practitioners in the county of Funen]. PMID- 14558387 TI - [Prescriptions of a new antibiotic following treatment with sulfamethizole or pivmecillinam]. PMID- 14558388 TI - [Sentinel node diagnosis in malignant melanoma. Reflections on relapse after histologically negative sentinel node]. PMID- 14558389 TI - [Clinical quality databases 2003. Handling of cases and evaluation of the year's applications]. PMID- 14558390 TI - [Picture of the month: notalgia paresthetica]. PMID- 14558391 TI - [Etiology and diagnostic methods in vocal cord paralysis]. PMID- 14558392 TI - [Toxic complacency]. PMID- 14558393 TI - [Dietary supplements should be taken seriously]. PMID- 14558394 TI - [A comment to the information from the National Board of Health and Welfare about piercing]. PMID- 14558395 TI - Enhancement of the results of neck liposuction with the FAMI technique. AB - Submental liposuction is an excellent procedure for improving the aging neck by reducing the cervicomental angle and increasing the definition of the mandibular border. However, it does not address the loss of volume of the chin and perioral areas caused by atrophy of hard and soft tissues. In this study, thirty patients underwent concomitant submental tumescent liposuction and fat augmentation with the FAMI (fat autograft muscle injection) technique in order to assess short-term results and complications associated with these two relatively noninvasive procedures. All patients had improvement in the cervicomental border, a smoother mandibular border, and a more proportioned chin following 3-5 days of swelling. Patients followed for 6 months to one year had long term retention of fat. There were no significant complications. In this preliminary series, submental tumescent liposuction with FAMI provided enhanced aesthetic results with little downtime and minimal complications. PMID- 14558396 TI - Non-surgical radiofrequency facelift. AB - There has been considerable interest in using non-ablative methods to rejuvenate the skin. The ThermaCool TC (Thermage Inc.) is a radiofrequency (RF) device that has been introduced to induce tightening of the address the problem of skin via a uniform volumetric heating into the deep dermis tightening, resulting in a 'non surgical facelift'. Radiofrequency produces a uniform volumetric heating into the deep dermis. Twenty treatment areas in 17 patients were treated to evaluate the efficacy and safety of RF treatment to the brow and jowls. The technique was found to produce gradual tightening in most patients, and there were no adverse effects. PMID- 14558397 TI - Photodynamic therapy with topical aminolevulinic acid and pulsed dye laser irradiation for sebaceous hyperplasia. AB - Successful and long-standing eradication of sebaceous hyperplasia has remained difficult due to the propensity of these lesions to be extensive. Current treatments include excision, electrodesiccation, laser vaporization, and oral isotretinoin, each often associated with unacceptable side effects or lesional recurrence. The purpose of this study was to evaluate the safety and effectiveness of laser-assisted photodynamic therapy using topical 5 aminolevulinic acid (5-ALA) and 595 nm pulsed dye laser (PDL) irradiation for the treatment of sebaceous hyperplasia. Ten patients with sebaceous hyperplasia received 1 or 2 treatments at 6 week intervals with topical 20% 5-ALA followed 1 hour later by 595 nm PDL irradiation. Matched lesions served as controls and were either treated with PDL alone or were left untreated. Patients were evaluated at regular intervals for 3 months. Results demonstrated that combination topical 5 ALA and PDL treatment effected better clinical results than PDL treatment alone. No changes were observed in untreated control lesions. Side effects were mild and limited to transient erythema, edema, and focal crusting. It is the conclusion of the authors that laser-assisted photodynamic therapy with topical 5-ALA and PDL irradiation can achieve safe and effective improvement of sebaceous hyperplasia. Further study is warranted to determine the longevity of the clinical results observed. PMID- 14558398 TI - Long-term sterility of fat frozen for up to 24 months. AB - Fat transfer is most effective when fresh fat is used, with excess fat frozen and saved for touch-ups. The obvious benefit to using frozen fat is that harvesting does not have to be done each time fat is injected. It is important to be able to easily store the fat in a freezer and be confident that it will not harbor bacterial growth. We studied frozen fat from 10 patients after 12 months and 10 patients after 24 months for sterility. Cultures were negative for all specimens, showing that frozen fat is safe to use and should not cause infection. PMID- 14558399 TI - Phosphatidylcholine in the treatment of localized fat. AB - Phosphatidylcholine was initially used in emergencies and in the treatment of atheroma plaques in cardiac diseases. Recently, it has also been used in the treatment of localized fat deposits. We report on the authors' clinical experience of the use of 250 mg/ml phosphatidylcholine injections in the treatment of subcutaneous fat deposits, showing the clinical response and side effects. Volunteers received phosphatidylcholine injections in several areas of localized fat deposits, with a minimum interval of one week and mean interval of 15 days between applications. Laboratory tests were performed during the period of the drug use. Clinical results reflect that phosphatidylcholine was efficacious in reducing the fatty pads in the treated areas, with few side effects. From the authors' point of view, the off-label use of phosphatidylcholine in the treatment of fatty pads and small areas of localized fat is safe, low cost, and effective. PMID- 14558400 TI - Treatment of hyperhidrosis. AB - Hyperhidrosis, or excessive sweating, is a disorder that may cause social isolation or occupational disability. It may be generalized or localized, and although frequently idiopathic it may be a manifestation of a number of important systemic diseases. Drugs, surgical procedures, and electrical devices may all be employed by the physician as therapeutic weapons to treat hyperhidrosis. PMID- 14558401 TI - Cosmeceuticals. Their role in dermatology practice. PMID- 14558402 TI - Skin cancer sites online: watch out for red flags. PMID- 14558403 TI - Yellow nail pigmentation following Depakote therapy. AB - Two months after being started on Depakote (divalproex sodium: Abbott Laboratories Inc., Abbott Park, Illinois), a 57-year-old female noticed the development of a transverse yellow band on all 20 proximal nails that eventually led to complete nail plate discoloration. Six to eight weeks after discontinuation of Depakote, normalization of her proximal nail plates was noted. Other anticonvulsants such as phenytoin and lithium have been documented to lead to nail pigmentation. Although several dermatologic reactions to Depakote have been described, we are not aware of any reported cases of nail discoloration secondary to Depakote. While our patient did have a history of renal disease with azotemia, we find it highly unlikely to be a contributing factor secondary to the fact that our patient's abnormality did not present similar to the nail abnormalities of renal disease. In conclusion, we found the temporal relationship between Depakote initiation/discontinuation and the nail discoloration to be highly indicative of Depakote as the source. Other anticonvulsants such as phenytoin and lithium have been documented to lead to nail pigmentation. Although several dermatologic reactions to Depakote have been described, we are not aware of any reported cases of nail discoloration secondary to Depakote. While our patient did have a history of renal disease with azotemia, we find it highly unlikely to be a contributing factor secondary to the fact that our patient's abnormality did not present similar to the nail abnormalities of renal disease. In conclusion, we found the temporal relationship between Depakote initiation/discontinuation and the nail discoloration to be highly indicative of Depakote as the source. PMID- 14558404 TI - Nephrogenic fibrosing dermopathy after five days of hemodialysis. PMID- 14558405 TI - Rosacea fulminans associated with pegylated interferon alpha-2B and ribavirin therapy. AB - Rosacea fulminans is characterized by the sudden onset of large, coalescing nodules and draining sinuses on the face. A few reports have linked medications to this condition, but none have described the onset of rosacea fulminans with pegylated interferon or ribavirin therapy. We report a patient who presented with rosacea fulminans after initiation of therapy for Hepatitis C. PMID- 14558406 TI - Morbilliform drug reaction with histologic features of pustular dermatosis associated with bryostatin-1. AB - Bryostatin-1 is a new chemotherapeutic agent that inhibits protein kinase C. The most common side effect and the dose limiting toxicity is myalgia. The cutaneous side effects reported during the phase I and II trials were alopecia, mucositis, nonspecific "rash," "bronzing," and hyperpigmentation in sun exposed areas. No specific acute drug eruptions have been reported. We present the first reported case of a morbilliform drug eruption with histologic features of intraepidermal and subcorneal spongiotic pustules containing eosinophils secondary to bryostatin 1. PMID- 14558407 TI - Using rituximab (anti-CD20 antibody) in a patient with paraneoplastic pemphigus. AB - Paraneoplastic pemphaigus (PNP) is a rare autoimmune mucocutaneous blistering disease that is commonly associated with underlying B-cell neoplasms. There is no standard therapy for PNP. Potent immunosuppression has been the only potentially effective treatment in the setting of malignancy because there is no correlation between tumor burden and activity of disease. Two recent case reports have noted the resolution of lesions of PNP after treatment of the underlying CD20+ B-cell lymphomas with rituximab. Rituximab is an anti-CD20 antibody that has had some success in treating proliferative B-cell disorders. We report a case of PNP in the setting of B-cell lymphoma that did not respond to this novel therapy, and discuss rituximab's putative mechanism of action along with the clinical settings in which this novel therapy may prove useful in the treatment of PNP. PMID- 14558409 TI - Statement on ethics and travel. PMID- 14558408 TI - Multiple fixed drug eruption with interferon/ribavirin combination therapy for hepatitis C virus infection. AB - Interferon/ribavirin combination therapy is the treatment of choice for chronic hepatitis C virus (HCV) infection. Cutaneous reactions to inferferon therapy, both local and distant from the injection site, are not uncommon. Eczematous and lichenoid reactions are most commonly reported. Interferon therapy has also been associated with the occurrence or worsening of a number of dermatologic disorders. We report the case of a 54-year-old man undergoing treatment with interferon/ribavirin combination therapy for hepatitis C who developed a multiple fixed drug eruption associated with interferon injections. To our knowledge, this is the first known reported case of a multiple fixed drug reaction triggered by interferon. PMID- 14558410 TI - The internet, a simple and convenient tool in Chlamydia trachomatis screening of young people. PMID- 14558411 TI - [A novel laboratory method of isolation and purification of tobramycin]. AB - A new laboratory method for isolation and purification of tobramycin by using extraction of a tobramycin derivative with benzaldehyde by methylene chloride, subsequent hydrolysis of azomethine and recrystallization of the formed tobramycin sulfate from solution of sulfuric acid in methanol was developed. The method allows to exclude the stage of chromatographic purification of tobramycin, to reduce the time of the process realization from 120-125 h to 15-20 h, to increase the yield of the target product from 37-40% to 60-65% without decreasing the product quality, to exclude a number of large-size and expensive equipment and to ensure high reproducibility of the technology. PMID- 14558412 TI - [Decrease of HIV-1 replicative capacity after serial passage in MT-4 cells under conditions of the combined antiretroviral therapy]. AB - The replicative activity of HIV-1/IIIB was determined in MT-4 cells under conditions of combined HIV reverse transcriptase and protease inhibitors in the presence of four different drug cocktails: (1) phosphazide, didanosine, nevirapine; (2) stavudine, didanosine, nevirapine; (3) phosphazide, didanosine, nevirapine, indinavir; (4) stavudine, didanosine, nevirapine, indinavir. The concentration of every inhibitor was 10 times higher than the 50% effective concentration. Alpha interferon was used as a natural antiretroviral agent in addition. The virus was subjected to 5 serial passages in the presence of the drug cocktails and then to 5 serial passages without the agents using cocultivation of infected and uninfected cells at ratio 1:5 to increase virus activity. Virus replication in the presence of all the drug combinations resulted in the appearance of HIV-1 variants with low replicative activity that was insignificantly increased during further passages even without the antiretroviral agents. If extrapolated to the clinical practice, these results indicate that all the drug cocktails were effective inhibitors of HIV-1 replication because the virus variants with high replicative activity did not emerge. Moreover, the results showed that the clinical use of the drug cocktails was promising. PMID- 14558413 TI - [Streptomyces griseolus # 182--a novel organism producing oligomycin antibiotics. Taxonomy, fermentation, and isolation]. AB - Target screening of natural immunosuppressors resulted in isolation of a strain of Streptomyces griseolus (No. 182) producing a complex of antifungal antibiotics. The strain proved to be an aerobe with the growth temperature of 26 to 28 degrees C. Morphological features and physiological properties of the strain were studied. Scanning electron microscopy revealed smooth, oval spores 1.10-1.25 mu in size. The findings showed that the strain belonged to Streptomyces griseolus. Unlike the previously described organisms producing the oligomycin complex the new strain formed straight or twisted sporophores and did not produce melanoid pigment or soluble pigment when grown on the Gauze mineral agar medium No. 1. The procedures for biosynthesis and chemical recovery of the antibiotic complex from the mycelium are described. The complex was shown to include 3 components at a ratio of 80:15:5 identified as oligomycins A, B and C respectively. The oligomycin complex was highly active against Aspergillus niger 137, Tolypocladium inflatum, Fusarium ocsisporum, Curvularia lunata 645 and Trichoderma alba F-32 (MIC 0.1-1.0 mcg/ml). The activity against yeast and bacterial cultures was observed only when the doses were higher than 100 mcg/ml. PMID- 14558414 TI - [Levofloxacin (Tavanic) in the treatment of corneal ulcers]. AB - The main clinical forms of cornea ulcers were systematized and high efficacy of levofloxacin in the treatment of bacterial ulcer of the cornea was shown. In the treatment of several bacterial affections of the eye the systemic use of levofloxacin should be combined with local antibiotic therapy. Combination of levofloxacin with ophthalmic drops of lomefloxacin (okacin) or ofloxacin (floxal) proved to be efficient. PMID- 14558415 TI - [Staphylococci and vancomycin--confrontation trends]. PMID- 14558416 TI - [Use of immunomodulators in viral infections]. PMID- 14558417 TI - [A novel marker of multiple drug resistance BCRP]. AB - Data on a new marker of multidrug resistance from the group of ABC-transporters- BCRP (breast cancer resistance protein) and in particular the data on the general characteristics of BCRP, spectrum of substrates for this transporter, data on the role of BCRP in human organism, pathways of inhibiting BCRP-dependent transport by pharmacological agents are presented. The data on BCRP expression in human normal tissues and tumor cells are considered in detail. Possible reasons of results inconsistency in evaluation of BCRP expression in different tumors and prognostic significance of BCRP in prediction of sensitivity to chemotherapy and aggressiveness of disease process are discussed. PMID- 14558418 TI - The road to safety. PMID- 14558419 TI - The kids' place. PMID- 14558420 TI - Branching out. PMID- 14558421 TI - Speeding ahead. PMID- 14558422 TI - Best feet forward. PMID- 14558423 TI - The job finders. PMID- 14558424 TI - From the drawing board. PMID- 14558425 TI - Stop before it starts. PMID- 14558426 TI - Stark revelations. PMID- 14558427 TI - If they were president.... PMID- 14558428 TI - [Appearance diagnosis No. 4. What is your diagnosis? Florid vitamin D deficiency rickets with secondary hyperparathyroidism]. PMID- 14558429 TI - [Office spirometry]. AB - Spirometry is a simple technique for assessment of pulmonary function. Analysis of airflow and lung volume recorded during standardized breathing maneuvers quantifies ventilatory limitation and classifies pulmonary function disturbances according to pathophysiological criteria. As an adjunct to clinical evaluation spirometry provides essential diagnostic and prognostic information for evaluation of cardio-pulmonary disorders, and for assessment of the course of lung diseases and their response to treatment. PMID- 14558430 TI - [Syncopes in children and adolescents]. AB - Syncopes are frequent in the pediatric age group. The majority of these events are benign and are so called neurocardiogenic syncopes. In a minority of affected children, a cardiac pathology is the underlying cause, these cases need to be recognized promptly as almost all of the underlying cardiac causes are potentially fatal conditions. Usually detailed history of syncopal events allows for differentiation between neurocardiogenic and other causes of syncope. In case the physical examination is normal, a simple ECG as the only additional laboratory test is adequate. Other laboratory tests are performed on the basis of suspicious findings in both history and physical examination. The typical neuricardiogenic syncopes rarely require medication in childhood, usually addressing some behavioural aspects with the patient are sufficient as only therapeutic measures. PMID- 14558431 TI - [Preoperative risk assessment and ambulatory management]. AB - Evidence and practical approach of Coronary Risk Assessment in noncardiac surgery are reviewed. Growing literature suggests a benefit of perioperative b-blockade. Therefore, risk stratification and management are described in algorithms in order to provide a useful tool for practitioners both in the hospital and ambulatory setting. PMID- 14558432 TI - [The interesting case from the pediatric polyclinic]. AB - An eleven year old girl from Turkey was diagnosed to have a periodic fever syndrome. The diagnosis of familial mediterranean fever was made by molecular analysis of a mutation in the MEFV-Gen which codes for pyrin. The disease is well known in the mediterranean area and belongs to the periodic fever syndromes. These syndromes are discussed for their differential diagnosis focused to childhood. PMID- 14558433 TI - Pharmacological comparison of the statins. AB - The statins (3-hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitors) represent drugs of first choice for treatment of hypercholesterolemia. The safety and efficacy of atorvastatin (CAS 134523-00-5), simvastatin (CAS 79902-63-9), lovastatin (CAS 75330-75-7), pravastatin (CAS 81093-37-0) and fluvastatin (CAS 93957-54-1) has been well documented. Statins decrease dose-dependently low density lipoprotein (LDL) cholesterol as well as coronary events and total mortality. Clinical outcome data indicate that for simvastatin the lowest number of treated patients is needed to prevent one major coronary event (NNT 15). Based on an approximately 30% reduction of LDL (valid surrogate parameter) atorvastatin (5 mg/day) and simvastatin (10 mg/day) are the most potent agents whereas 40 mg of lovastatin or pravastatin and 60 mg of fluvastatin are needed to reach this "therapeutic target". While all statins share the same mode of action their pharmacokinetic properties and their susceptibility to drug interactions differ slightly. Agents inhibiting CYP3A4 (e.g. grapefruit juice, itraconazole, cyclosporine) should be discouraged if a patient is on atorvastatin, lovastatin or simvastatin. Likewise, fluconazole interferes with the CYP2C9-mediated hepatic elimination of fluvastatin. Moreover, coadministration of gemfibrozil should be avoided because it seems to increase the very low risk for statin-induced rhabdomyolysis. Several statins are available and their equieffective doses have been defined. Selection of a particular drug should be primarily based on clinical outcome data. However, costs and in certain situations the pharmacokinetic profile including the interaction potential of the statins should be taken into account. PMID- 14558434 TI - Pharmacokinetics of the new pyrimidine derivative NS-7, a novel Na+/Ca2+ channel blocker. 1st communication: plasma concentrations and excretions after a single intravenous 14C-NS-7 injection to rats, dogs and monkeys. AB - Plasma concentration profiles and excretion were investigated after a single intravenous injection of 14C-NS-7 (4-(fluorophenyl)-2-methyl-6-(5 piperidinopentyloxy)pyrimidine hydrochloride, CAS 178429-67-9), a novel Na+/Ca2+ channel blocker, to rats, dogs and monkeys. Plasma protein binding of this drug was determined in vitro and in vivo. AUC0-infinity values for radioactivity and NS-7 after the intravenous administration of 14C-NS-7 to male rats increased with the dose, namely from 0.04 to 5 mg/kg (radioactivity) and from 0.2 to 5 mg/kg (NS 7), indicating the linearity of the drug's pharmacokinetics. Plasma concentrations of the unchanged drug after the intravenous injection of 0.2 mg/kg 14C-NS-7 decreased biexponentially, respective t1/2 beta values being 15.9 h in the male and 22.4 h in the female rats. The t1/2 beta values difference in the males and females might be due to sex differences in NS-7 metabolism. Urinary and fecal excretions of radioactivity within 168 h of administration were 33.0 and 61.4% of the dose in the male and 35.0 and 53.2% in the female rats. No radioactivity was detected in air exhaled from the males and females collected for 168 h after NS-7 administration. Within 24 h of administration, respective biliary excretions for the male and female rats were 26.1 and 11.9% of the dose. Of this excreted radioactivity, 34.9% was reabsorbed in the males. NS-7 plasma concentrations decreased biexponentially after intravenous administration of 0.2 mg/kg 14C-NS-7 to dogs and monkeys. The elimination half-life was 18 h for the dogs and 9.52 h for the monkeys. Urinary and fecal excretions of radioactivity within 168 h of administration were 24.2 and 70.0% of the dose for the dogs, and 63.3 and 24.8% for the monkeys. These species differences in excretion may be due to differences in urinary metabolite compositions. In vitro protein binding of NS 7 showed no marked species differences and was independent of the NS-7 concentration. Binding of 14C-NS-7 in the sera of rats, dogs, monkeys and humans was 90.7%, 73.5% 79.0% and 87.1%, respectively. Binding to human serum albumin, alpha 1-acid glycoprotein and lipoprotein was 56.2%, 45.4% and 79.5%, in the range of 4-40 ng/ml. In vivo binding in rat serum 5 min, 6 h and 24 h after the intravenous injection of 14C-NS-7(0.2 mg/kg) ranged from 89.6 to 90.6%. PMID- 14558435 TI - Incidence of pain after intravenous injection of a medium-/long-chain triglyceride emulsion of propofol. An observational study in 1375 patients. AB - BACKGROUND AND OBJECTIVE: To assess incidence and intensity of pain on intravenous injection of propofol (CAS 2078-548) in an emulsion of medium chain/long-chain triglycerides (MCT/LCT, 50:50) in patients undergoing different elective surgical interventions. METHODS: The new solvent was used for induction of general anesthesia. Spontaneous pain reactions and pain elicited upon questioning were assessed. Patients were asked to grade the pain as mild, moderate or severe. Co-medication with sedative or analgesic drugs, size of the intravenous cannulae, site of injection and administration as a single bolus or in divided doses were recorded. RESULTS: Overall incidence of pain was 28.4% (390 out of 1375 patients). Twelve percent of the patients complained spontaneously and 16.4% reported pain after questioning. Pain intensity was graded as mild by 16.7% of the patients. The incidence of pain was significantly less when using an antecubital vein compared with a forearm or dorsal hand vein (p = 0.017 spontaneously reported pain, p = 0.001 pain elicited upon questioning). The number of patients complaining spontaneously of pain was significantly lower (p = 0.006) for large size than for small and medium size cannulae. CONCLUSIONS: The incidence of pain on injection of a medium-/long-chain triglyceride propofol formulation was 28.4% with 16.7% of the patients reporting mild pain. The use of an antecubital vein or a large size venous cannula appears to reduce the injection pain. PMID- 14558436 TI - Single oral dose pharmacokinetic interaction study of manidipine and delapril in healthy volunteers. AB - OBJECTIVE: The objective of the study was to assess potential pharmacokinetic interactions between delapril, an angiotensin conversion enzyme inhibitor, and manidipine, a calcium channel antagonist, prior to the development of a fixed combination drug product. METHODS: Eighteen healthy male volunteers received a single oral dose of 10 mg manidipine dihydrochloride (CAS 89226-75-5), or 30 mg delapril hydrochloride (CAS 83435-67-0), or both simultaneously, according to a fully balanced three-way cross-over design. The three treatments were separated by a one-week washout period. Blood samples were collected during 24 h for plasma determination of manidipine and metabolite M-XIII and/or of delapril and metabolites M1, M2 and M3, using specific LCMS/MS methods. RESULTS: The bioavailability of manidipine and M-XIII was slightly decreased by concomitant administration of delapril (manidipine: Cmax-19% and AUC infinity-11% M-XIII: Cmax-17% and AUCt-18%). The bioavailability of delapril was not influenced by co administration with manidipine (Cmax-7% and AUC infinity +4%). The effect on delapril pharmacologically active metabolites M1 and M3 was negligible. The inactive metabolite M2 underwent a 13% reduction of Cmax and AUC infinity. The 90% confidence intervals were confined within limits of acceptance (70-143% for Cmax and 80-125% for AUC). Mean residence times and apparent elimination half lives were unaltered. Blood pressure and heart rate versus time profiles were similar during the three treatments. CONCLUSIONS: Simultaneous oral administration of 10 mg manidipine and 30 mg delapril does not significantly alter the pharmacokinetics of either drug or that of their principal metabolites. PMID- 14558437 TI - Relative bioavailability and bioequivalence study of theophylline sustained release formulations. AB - The objective of this study was to determine the bioequivalence of two theophylline (CAS 58-55-9) sustained release formulations containing 400 mg (Theophyllin 400 retard Heumann, formulation A) and 375 mg (formulation C) theophylline, respectively. In addition, the relative bioavailability of the sustained release formulations in comparison to an oral solution (formulation B) was investigated. Twenty-four healthy male volunteers participated in the open randomized three-way crossover study. Multiple doses of the formulations were administered during three study periods of four days each (A: 400 mg once daily; B: 133 mg t.i.d.; C: 375 mg once daily). The absorption kinetics and the bioavailability of theophylline were investigated by model-independent and deconvolution methods. The relative bioavailability of formulation A as compared to the solution was 72%. The oral sustained release capsules did not exhibit any differences with respect to AUCss, tau and Css, max whereas differences were detected regarding tss, max and peak trough fluctuation indicating minor deviations of the plasma profiles of both formulations. However, 90% confidence intervals of the ratios of AUCss, tau and Css, max were within the respective acceptance limits. Thus, both formulations are bioequivalent considering rate and extent of absorption. PMID- 14558438 TI - Synthesis and anti-inflammatory activity of alkyl/arylidene-2-aminobenzothiazoles and 1-benzothiazol-2-yl-3-chloro-4-substituted-azetidin-2-ones. AB - Ten new derivatives of 1-benzothiazol-2-yl-3-chloro-4-substituted-azetidin-2-ones (3a-j) were synthesized using various Schiff bases (alkyl/arylidene-2 aminobenzothiazoles; 2a-j), which in turn were prepared starting from 2 aminobenzothiazole (1). All the synthesised compounds were characterised by elemental analyses and spectral (IR, 1H-NMR, 13C-NMR and EI-MS) data. The title compounds 2a-j and 3a-j were screened in vivo using carrageenan-induced rat paw edema model. All the test compounds showed anti-inflammatory activity when tested in vivo. In general, compounds 3a-j were found to be more potent compared to compounds 2a-j. Among the compounds tested, compound 2g in the alkyl/arylidene-2 aminobenzothiazoles series and compound 3 g in the 1-benzothiazol-2-yl-3-chloro-4 substituted-azetidin-2-ones series were found to be the most potent. All the test compounds were also evaluated to check the gastric ulcer incidence. In gastric ulceration studies, all the test compounds were generally found to be safe at the 100 mg/kg dose level. Furthermore the most potent compounds 2 g and 3 g from each series were screened in vitro for inhibition of both COX-2 and COX-1 catalysed prostaglandin biosynthesis (radiochemical assay). Like most of the commercially available non-steroidal anti-inflammatory drugs (NSAIDs), in the in vitro study, compounds 2 g and 3 g showed anti-inflammatory activity by blocking the metabolism of arachidonic acid to prostaglandin via the cyclooxygenase pathways. In general, in the vitro assay, test compounds 2 g and 3 g were found to be more active after 15 min pre-incubation with the enzyme. Compound 3 g was found to be more COX-2 selective, while compound 2 g was found to be equally COX-2 and COX-1 selective. PMID- 14558439 TI - Prevention of islet allograft rejection in diabetic mice by targeting Janus Kinase 3 with 4-(4'-hydroxyphenyl)-amino-6,7-dimethoxyquinazoline (JANEX-1). AB - Janus kinase (JAK) 3-deficient mice were not able to reject allogeneic islet allografts. The JAK3 inhibitor 4-(4'-hydroxyphenyl)-amino-6,7 dimethoxyquinazoline (CAS 202475-60-3, JANEX-1, WHI-P131) prevented the rejection of islet allografts in mice with a normal JAK3 expression status. The combination of JANEX-1 and cyclosporin A (CAS 59865-13-3) was more effective than either agent alone. PMID- 14558441 TI - Pharmacokinetics of ciprofloxacin in healthy Mexican volunteers. AB - Ciprofloxacin (CAS 85721-33-1) is a gyrase inhibitor used against a wide number of bacteria which develops a very low bacterial resistance. Its pharmacokinetics has been extensively studied using two main methods, liquid chromatography and microbiological assay. The pharmacokinetics of ciprofloxacin administered orally was evaluated in healthy Mexican volunteers after a dose of 500 mg. Ciprofloxacin was assayed from plasma by a specific HPLC method reading absorbance at 280 nm. Pharmacokinetic parameters were similar to reported values, including a previous study on Mexican healthy volunteers with the microbiological assay. AUC was 12.11 mg h/l, Cmax 2.44 mg/l, tmax 0.79 h and half-life 3.8 h. tmax was slightly shorter than those in other studies. Ciprofloxacin presented no adverse reactions and can be dosed to people with Amerindian origin in the same dose regimes as prescribed to Caucasians in order to achieve minimal inhibitory concentrations against a wide range of microbial pathogens. PMID- 14558440 TI - Synthesis and evaluation of novel 7-trifluoromethyl-4-(4-substituted anilino)quinolines as antiparasitic and antineoplastic agents. AB - Several novel derivatives bearing the 7-trifluoromethyl-4-(4-substituted anilino) quinoline skeleton were synthesized and evaluated for their in vitro activity against the blood streaming form of the parasites Trypanosoma brucei rhodesiense, the trypomastigotes of Trypanosoma cruzi cultured in rat skeletal myoblasts, the amastigotes form of Leishmania donovani, Plasmodium falciparum (K1 strain) infected erythrocyte suspension, as well as their toxicity towards rat skeletal L 6 cells. In addition, three of the synthesized compounds were tested for their in vitro antitumor activity towards 60 human tumor cell lines by the National Cancer Institute (NCI). Compound 1-(4-[[7-(trifluoromethyl)quinolin-4 yl]amino]phenyl)ethan-1-one thiosemicarbazone 23 exhibited potential activity against T. b. rhodesiense, T. cruzi and P. falciparum with IC50's of 0.278, 0.85 and 0.417 microgram/ml, respectively. These values are even more valuable since this compound was clearly non-toxic (cytotoxicity IC50 > 90 micrograms/ml), leading to in vitro therapeutic indices of > 323, > 106 and > 216, respectively. Meanwhile, compound N-[4-[5-(4-chlorophenyl)-4,5-dihydroisoxazol-3-yl]phenyl]-7 (trifluoromethyl) quinolin-4-amine 21 showed broad spectrum antitumor activity with full panel median growth inhibition GI50 (MG-MID) of 1.95 mumol and total growth inhibition TGI (MG-MID) of 6.87 mumol, respectively. Compound 23 represents a very good prospective as a lead compound able to combat three tropical diseases at a time. However, the pattern for the antitumor activity did not parallel any of the antiparasitic ones, indicating that non-common mechanisms of action may exist among these activities. PMID- 14558442 TI - In vitro antifungal activity of N,N-phenyl-1,2,3,4-thiatriazole-5-yl-2,4-beta resorcylcarbothioamide. AB - The aim of this study was the determination of the antifungal activity of N,N phenyl-1,2,3,4-thiatriazol-5-yl-2,4-beta-resorcylcarbothioamide (PTR) against Candida albicans, non-albicans Candida species, dermatophytes and moulds and of its influence on the enzymatic activity of C. albicans strains. The reference strains C. albicans ATCC 10231, 200 of C. albicans strains, 7 of non-albicans C. species, 12 dermatophyte strains and 20 mould strains were isolated from different patients. The mean minimum inhibitory concentration (MIC) of PTR against C. albicans strains isolated from patients was 19.6 mg/l, for reference C. albicans ATCC 10,231 it was 12.5 mg/l on Sabouraud's medium (SB). The mean MIC of isolates from patients was 16.9 mg/l, and reference strains 6.25 mg/l on YNB medium, respectively. The MIC of PTR against 7 non-albicans C. species was 27.7 mg/l on SB and 15.6 mg/l on YNB, respectively. The mean MIC of PTR against C. albicans strains isolated from patients was 14.9 mg/l, C. albicans ATCC 10,231 6.25 mg/l and non-Candida species strains 14 mg/l on RPMI medium. The MICs of PTR against dermatophytes ranged from 3 to 25 mg/l. The MICs of PTR against moulds were 25 mg/l and 100 mg/l, respectively. PTR inhibited the enzymatic activity of selected hydrolases of C. albicans and non-Candida species strains. PTR exerts a potent antifungal activity against the yeast-like fungi strains, moulds and dermatophytes. This new compound inhibited the enzymatic activity of selected hydrolases. PMID- 14558443 TI - Introductory conference: on the relevance of protected areas for the research on conservation ecology: from fundaments to applications. AB - Regarding the present rate of species extinction, preservation of biodiversity has become a prerequisite to achieve sustainable development. Among major challenges an urgent need in research in biological conservation is required to reach this goal. Protected areas deserve special importance due to their relevance for development of research both in fundamental and applied ecology. This papers aims at making a critical review of possibilities and priorities in conservation ecology in these areas. PMID- 14558444 TI - The relationships between taxonomy and conservation biology in the century of extinctions. AB - The 21st century will be that of the crisis of biodiversity and of extinctions: a majority of the planet's species might disappear before having even been studied by man. The discipline in charge of this study, taxonomy, is currently facing severe problems, concerning its status inside biology and its relationships with conservation biology. Yet taxonomy and conservation biology would have much to gain to becoming allies rather than largely foreign disciplines. This would require some important changes in mentality and behaviour. PMID- 14558445 TI - Genetics and conservation biology. AB - Conservation genetics encompasses genetic management of small populations, resolution of taxonomic uncertainties and management units, and the use of molecular genetic analyses in forensics and to understanding species' biology. The role of genetic factors in extinctions of wild populations has been controversial, but evidence now shows that they make important contributions to extinction risk. Inbreeding has been shown to cause extinctions of wild populations, computer projections indicate that inbreeding depression has important effects on extinction risk, and most threatened species show signs of genetic deterioration. Inappropriate management is likely to result if genetic factors are ignored in threatened species management. PMID- 14558446 TI - Implications of an individualistic lifestyle for species conservation: lessons from jealous beasts. AB - Polecat populations show a very low genetic diversity and a high inbreeding coefficient. Furthermore, the estimate of effective population size is alarmingly low. Polecats Mustela putorius populations are structured into scattered breeding sub-units usually made up of one male and two females, according to a polygynous mating system. Because a strict spatio-temporal segregation was observed between males and females, we propose to call individualistic such species. We suggest that the solitary habits of individualistic species may result in or worsen a high inbreeding and exacerbate their conservation issue, a crucial perspective for critically endangered species such as the European mink. PMID- 14558447 TI - Genetic diversity in European red deer (Cervus elaphus L.): anthropogenic influences on natural populations. AB - Allozyme, microsatellite and mtDNA (RFLP and sequence) data of European red deer populations were examined as to their capability of indicating anthropogenic influences such as the keeping of animals in enclosures, selective hunting for trophies translocation of specimens to improve trophy quality and habitat fragmentation. Deer in enclosures revealed considerable deviations of allele frequencies from isolation-by-distance expectations but no remarkable loss of genetic diversity. Particular allozyme genotypes were associated with antler morphology, and selective hunting was shown to alter allele frequencies in the expected direction. Habitat fragmentation is reflected by various kinds of genetic markers but due to the lack of information on population histories no unequivocal evidence on particular human activities could be obtained. PMID- 14558448 TI - AFLP: a useful tool for biodiversity conservation and management. AB - The access to new molecular markers is not always possible for all researchers, due to limited laboratory and economical resources. A new technique, called AFLP, seems to be a simple and robust method to obtain a large number of variable molecular markers. In this article I show the results of AFLP analysis in three different conservation projects requiring different level of genetic variability. I also compare the multi-locus AFLP results with single-locus markers (microsatellites) and haploid organellar marker (mtDNA) sequences. The results suggest that the AFLP technique could be very useful in a wide range of conservation studies. PMID- 14558449 TI - Molecular survey of genetic diversity in the endangered European mink Mustela lutreola. AB - The European mink Mustela lutreola is regarded as one of the most endangered mammals in the world. We chose to characterize microsatellite loci in order to investigate the pattern of decline of this species. We used primer pairs developed for a related species Mustela vison to genotype individual of Mustela lutreola. Out of 19 primer pairs used 8 were useful for our purpose. The conservation of primer sequence point out the problem of neutrality of some microsatellite loci as this conservation could be related to strong selection pressure on those loci. Finally we present the first data allowing an estimation of heterozygosity of French population of European mink. PMID- 14558451 TI - Evaluation of methodology for detection of genetic bottlenecks: inferences from temporally replicated lake trout populations. AB - Statistical methods have been proposed to detect recent bottlenecks on the basis of genetic characterizations of natural populations. In the absence of direct estimates of contemporary or historical population numbers, we tested the indirect M-ratio method based on microsatellite motif size frequency profiles using three historical and three contemporary Great Lakes populations of Salvelinus namaysuch for which severe reductions in population numbers are documented. Simulations employing plausible ranges of empirical population parameter values were used to explore bottleneck likelihood surfaces. We show that single values of the M-ratio are not sufficient to unambiguously infer a bottleneck without knowledge of mutation rates and effective population size (i.e., 4Ne mu or [symbol: see text]). Inferences of the degree of population bottleneck would be best supported if analyses were conducted across plausible ranges of [symbol: see text] and by qualitative comparisons among population samples. PMID- 14558450 TI - Conservation genetics of carnivores in Italy. AB - Pleistocene climatic changes shaped the patterns of biodiversity in Europe and around the Mediterranean. Describing the phylogeographic structure of animal populations and inferring past population dynamics is essential to develop a framework for conservation biology in Europe. Direct persecution, habitat loss, population fragmentation and hybridization with domesticated conspecifics, are the main threats to the survival of large mammalian species. In this paper I will summarize the available information on phylogeography and population genetics of brown bear, wolf, wildcat and otters in Italy and in Europe. PMID- 14558452 TI - Inbreeding and road effect zone in a Ranidae: the case of Agile frog, Rana dalmatina Bonaparte, 1840. AB - Inbreeding has often been invoked in the extinction of local populations. In eleven western France populations of Agile frog studied, observed heterozygosity was significantly lower than expected in all cases, giving new evidence of such a depression in small populations. It especially occurred in ponds located near an highway rather than in undisturbed populations (FIS = 0.544 and 0.315, respectively). Thus, our results argue for a "road effect zone". Discussing about road distance and conservation policies, we propose that roads are directly involved in inbreeding and in local extinction. Thus, road construction ought to consider conservation management. PMID- 14558453 TI - Phylogenetics and the conservation of a diverse and ancient flora. AB - Phylogenetics has a valuable role in the conservation of the flora of Western Australia particularly in the south-west where the complex evolutionary history has resulted in a diverse flora with both relictual and recently evolved components. Phylogenetic analysis contributes to conservation of plants through clarification of taxonomic status, identification of unique evolutionary lineages, determination of relictual and recently derived species, determination of phylogenetic value for conservation priority setting, and identifying phylogenetically independent comparisons between rare and widespread species. The contribution of phylogenetic analysis to these aspects of effective conservation management are discussed with examples from various species in Western Australia. PMID- 14558454 TI - Hare populations in Europe: intra and interspecific analysis of mtDNA variation. AB - We report results of a project aimed at analysing genetic diversification in Lepus corsicanus (italian hare), carried out at the INFS as part of an ongoing project on the biology of L. corsicanus. Samples of Italian hare were collected in different localities in central-southern Italy and Sicily. Moreover samples of L. europaeus (brown hare), as well as other species of the genus Lepus, were collected in Italy and Europe in order to define the phylogenetic relationships of the Italian hare. Results show a clear genetic distinction of the Italian hare. Consequently the status of true species is warranted at the molecular level. Phylogenetic relationships among species suggest the presence of two distinct groups, the Italian hare belonging to an evolutionary lineage including species which probably originated in a glacial refugia in southern Europe before colonization by the brown hare. On the other hand the brown hare belongs to a different lineage including species of African origins. PMID- 14558455 TI - Frog alien species: a way for genetic invasion? AB - European water frogs are characterized by anthropic introductions and Rana ridibunda may be considered as an invasive species. As such translocations may result in introgression of exotic genes in native populations, i.e. genetic pollution, we studied genetic characteristics (on 11 allozymic loci) of natural versus introduced water frogs. Our study contributed to (1) disclose 3 genetic markers allowing the identification of exotic frogs; (2) quantify the proportion of exotic frogs found in natural populations; and (3) suggest how genetic pollution may arise in these frogs. PMID- 14558456 TI - Applying molecular genetic tools to the conservation and action plan for the critically endangered Far Eastern leopard (Panthera pardus orientalis). AB - A role for molecular genetic approaches in conservation of endangered taxa is now commonly recognized. Because conservation genetic analyses provide essential insights on taxonomic status, recent evolutionary history and current health of endangered taxa, they are considered in nearly all conservation programs. Genetic analyses of the critically endangered Far Eastern, or Amur leopard, Panthera pardus orientalis, have been done recently to address all of these questions and develop strategies for survival of the leopard in the wild. The genetic status and implication for conservation management of the Far Eastern leopard subspecies are discussed. PMID- 14558457 TI - Conservation of endangered species and the patterns and propensities of biodiversity. AB - It is commonly asserted in the ecological and economic literature that habitat loss is the main cause of loss of imperiled species. The evidence clearly shows that habitat loss is a common contributing factor, but there is little evidence that it is the most important factor. Studies that have focused on the mechanisms of species loss have failed to produce models capable of predicting patterns of loss as a function of human activities. I propose that this is because ecologists have employed an unrealistic conceptual model of the functioning of natural systems. Karl Popper's construct of the propensities of natural systems provides a more realistic view, and better potential to yield predictive models. I provide two examples of patterns of biodiversity and species loss in Canada where mechanistic reasoning is inconsistent with the observed propensities of species loss. PMID- 14558458 TI - Challenges to reestablishment of free-ranging populations of black-footed ferrets. AB - The black-footed ferret (Mustela nigripes) of North America is critically endangered due in part to its extreme specialization on formerly stable and abundant prairie dogs (Cynomys). Its close relative, the Siberian polecat (M. eversmannii) seems to have been subjected to a varying environment that was not conductive to specialization. One source of environmental variation in Asian steppes was plague (caused by Yersina pestis), which was absent from North America. Introduction of plague to North America presents serious challenges to ferret recovery. Partial solutions to other biological and political problems have been found, resulting in improved production in captivity, increased survival post-release, and thriving populations in plague-free South Dakota. PMID- 14558459 TI - Research needs for lion conservation in West and Central Africa. AB - The lion has historically probably been widespread at low densities in West and Central Africa, nowadays they are largely restricted to small isolated populations inside protected areas. The total number is probably between 1200 and 2700, the best possible guesstimate would be 1700. Mankind is the main cause for the suspected decline of lion populations, both inside and outside protected areas. Very little research has been done on West and Central African lions a few examples are summarized here. The international community is slowly becoming aware of threats to lions in the region and some initiatives for lion conservation have started. PMID- 14558460 TI - Count data, detection probabilities, and the demography, dynamics, distribution, and decline of amphibians. AB - The evidence for amphibian population declines is based on count data that were not adjusted for detection probabilities. Such data are not reliable even when collected using standard methods. The formula C = Np (where C is a count, N the true parameter value, and p is a detection probability) relates count data to demography, population size, or distributions. With unadjusted count data, one assumes a linear relationship between C and N and that p is constant. These assumptions are unlikely to be met in studies of amphibian populations. Amphibian population data should be based on methods that account for detection probabilities. PMID- 14558461 TI - Biodiversity in salt marshes: from patrimonial value to ecosystem functioning. The case study of the Mont-Saint-Michel bay. AB - Until 1979, European salt marshes were known only through the inventories of fauna and especially of flora. On such criteria, the salt marshes of the Mont Saint-Michel bay (France) were regarded as most significant of the French coasts. However, it took 20 years of research on the role of these wetlands of the estuaries-salt marsh systems to highlight the ecological, social and economic interest of this ecotone, between continental and marine systems, a long time considered as territory "without value", except for stock breeders or hunters. PMID- 14558462 TI - Habitat fragmentation and amphibian conservation: building a tool for assessing landscape matrix connectivity. AB - We have attempted modelling biological connectivity for a toad population in a floodplain in the combined framework of population dynamics and landscape ecology. We took advantage of the GIS package for establishing friction maps for the focal species by attributing to each habitat type a resistance index. This approach made it possible to draw migration zones around each pond the area and the shape of which depended on landscape structure. We included mortality from road traffic by assigning a virtual population to each pond. The resulting map shows the potentiality of such an approach in the assistance of landscape management. PMID- 14558463 TI - Shorebird community variations indicative of a general perturbation in the Mont Saint-Michel bay (France). AB - The Mont-Saint-Michel bay located on the East Atlantic Flyway is the first site in France for wintering shorebirds, with, on average, 53,000 individuals in January. Seven species represent 96% of that community: dunlin (Calidris alpina), knot (Calidris canutus), oystercatcher (Haematopus ostralegus), curlew (Numenius arquata), grey plover (Pluvialis squatarola), bar-tailed godwit (Limosa lapponica) and black-tailed godwit (Limosa limosa). The international bird census organised by Wetlands International in mid-January gave us the opportunity to study, for a 23 years period, population variations in the bay. Despite a quite good carrying capacity, we showed that the decreasing proportion of 4 species in the bay relative to the other French populations may indicate a general perturbation of the bay. We discuss the different hypotheses to explain that observation. PMID- 14558464 TI - Sheep grazing as management tool in western European saltmarshes. AB - The effects of sheep grazing on plant community structure and diversity were studied in saltmarshes of the Mont-Saint-Michel bay. This study took place at two scales: (1) at the scale of the entire bay to explore the changes in plant community over a ten year period; and (2) locally with the use of experimental exclosure set up to mimic the abandonment of grazing. Moderate grazing generally enhanced plant richness and diversity, while the absence of grazing and overgrazing lead to a decrease in diversity and richness. The development of management strategies is becoming critical to preserve the diversity of saltmarshes functions. PMID- 14558465 TI - Reintroducing antelopes into arid areas: lessons learnt from the oryx in Saudi Arabia. AB - We focus on constraints faced by antelopes reintroductions in arid environments, and propose keys to enhance their success, using the oryx project in Saudi Arabia as example: (1) Monitoring and management of reintroduced populations appear more important than the number of released animals; (2) Because of the low accuracy of population size estimators, we recommend to implement a continuous monitoring and to use several estimators to assess the reintroduced population size; (3) Reintroduction schedule should take into account the unpredictability of food resources in arid environments; (4) The re-establishment of desert antelopes depends as a priority on the enforcement of regulations to avoid poaching. PMID- 14558466 TI - Trends of a bitterling (Rhodeus sericeus) population in a man-made ditch network. AB - This study describes the trends of a bitterling population in the Bourgneuf Marsh in relation to human activities. Although there were no bitterling during the period 1987-91, the species rapidly colonized the ditch network sampled (density reaching 61.7 +/- 47.0 individuals/100 m2 and frequency of occurrence around 70 75%) during a 5-year period (1997-2001). Clearance operations probably favoured the expansion of this vulnerable fish species because of positive associations of bitterling with large and deep ditches that had been cleared in the last 2-3 years and characterized by a small cover of submerged plants. Management recommendations are discussed for conserving the bitterling population. PMID- 14558467 TI - Which future for the French Pyrenean brown bear (Ursus arctos) population? An approach using stage-structured deterministic and stochastic models. AB - The Pyrenean brown bear (Ursus arctos) population is considered as one of the most seriously threatened with extinction in Western Europe. To assess its viability and possible needs of augmentation, we develop deterministic and stochastic stage-structured demographic models. The deterministic model reveals that a bear population cannot have a high annual growth rate and is particularly sensitive to breeder survival. High demographic parameters appear to be crucial to population persistence, especially for a small population that remains vulnerable to demographic and environmental stochasticities. The Pyrenean population cannot therefore be considered as viable. Successful conservation strategies for this population would require releasing more bears in both sub populations in the near future. PMID- 14558468 TI - Sustainable management of fixed dunes: example of a pilot site in Brittany (France). AB - The sand-dunes of Quiberon was chosen as a pilot site to investigate experimentation in conservatory management. Sand burial is necessary to conserve the semi-fixed dune which is a transitory dynamic stage. In the fixed dune, low disturbances benefit the vegetation diversity while heavy ones create serious injury. An opening of the milieu can restore very fast but a naked substrate is difficult to heal. The deposition of gorse branches is then efficient to facilitate the restoration. The fixed dune biodiversity is linked to human activities. Disturbances, natural or not, may be used as management tools. PMID- 14558469 TI - Beaver lodge location on the upstream Loire River. AB - In the part of the Loire River recently colonized by Eurasian beavers, we compared habitat characteristics among sites with lodges, sites with cut trees and sites without beaver. The absence of sandbank and canopy cover (by 10-15-m tall trees, by tall Salicaceae, and by bushy Salicaceae) appeared as good predictors for lodge settling. Based on this model, the number of proper lodge sites was estimated for the next downstream 36 kilometers stretch. The number of favourable sites decreases as anthropization increases. PMID- 14558470 TI - Effect of habitat fragmentation on dispersal in the butterfly Proclossiana eunomia. AB - Comparison of dispersal rates of the bog fritillary butterfly between continuous and fragmented landscapes indicates that between patch dispersal is significantly lower in the fragmented landscape, while population densities are of the same order of magnitude. Analyses of the dynamics of the suitable habitat for the butterfly in the fragmented landscape reveal a severe, non linear increase in spatial isolation of patches over a time period of 30 years (i.e. 30 butterfly generations), but simulations of the butterfly metapopulation dynamics using a structured population model show that the lower dispersal rates in the fragmented landscape are far above the critical threshold leading to metapopulation extinction. These results indicate that changes in individual behaviour leading to the decrease of dispersal rates in the fragmented landscape were rapidly selected for when patch spatial isolation increased. The evidence of such an adaptive answer to habitat fragmentation suggests that dispersal mortality is a key factor for metapopulation persistence in fragmented landscapes. We emphasise that landscape spatial configuration and patch isolation have to be taken into account in the debate about large-scale conservation strategies. PMID- 14558471 TI - La Reunion and Mayotte cockroaches: impact of altitude and human activity. AB - Competitive displacement is the outcome of interspecific competition between invading and native species. Alien insect species invade not only cultivated areas but also natural habitats. We prospected the cockroach fauna on two oceanic islands (Mayotte and La Reunion) to investigate the impact of altitude and anthropic disturbance on the distribution of these species. Most invading cockroach species seem to benefit from cultivation of land whereas endemic species occupy more specific and reduced habitats that are threatened by human impact. PMID- 14558472 TI - The global allostery model of hemoglobin: an allosteric mechanism involving homotropic and heterotropic interactions. AB - Studies of the allosteric mechanism of hemoglobin (Hb) have evolved from phenomenological descriptions to structure-based molecular mechanisms, as the molecular structures of Hb in deoxy and ligated states have been elucidated. The MWC two-state concerted model has been the widely accepted as the most plausible of the allosteric mechanisms of Hb. It assumes that the O2-affinity of Hb is regulated/controlled primarily by the T/R quaternary structural transition and that heterotropic effectors bind preferentially to T (deoxy) Hb to shift the T/R allosteric equilibrium toward the T state. However, recent more comprehensive O2 binding measurements of Hb have revealed a new mechanism, the Global Allostery model. It describes that the O2-affinity and the cooperativity are modulated in greater extents and the Bohr effect is generated primarily by the tertiary structural changes in both T (deoxy) and R (ligated) states of Hb. Differential interactions of heterotropic allosteric effectors with both T (deoxy) and R (ligated) states of Hb induce these tertiary structural changes. The X-ray structure of a complex of R (ligated) Hb with BZF, a potent heterotropic effector, has revealed the stereo-chemical influence of these effectors on the structure of R (ligated) Hb, resulting in the reduction of the ligand affinity in R (ligated) Hb. This model stresses the fundamental importance of the heterotropic interactions in regulation/control of the functionality of Hb. They alter the tertiary structures of both T (deoxy) and R (oxy) Hb, leading to large scale modulations of the O2 affinity (KT and KR), and consequently the cooperativity (KR/KT) and the Bohr effect (delta P50/delta pH) from a global viewpoint of allostery in Hb. PMID- 14558473 TI - Interaction of nitrogen monoxide with hemoglobin and the artefactual production of S-nitroso-hemoglobin. AB - Hemoglobin (Hb) is probably the most thoroughly studied protein in the human body. However, it has recently been proposed that in addition to the well known function of dioxygen and carbon dioxide transporter, one of the main roles of hemoglobin is to store and transport nitrogen monoxide. This hypothesis is highly disputed and is in contrast to the proposal that hemoglobin serves as an NO. scavenger in the blood. In this short review, I have presented the current status of research on the much-debated mechanism of the reaction between circulating hemoglobin and NO.. Despite the fact that oxyHb is extremely rapidly oxidized by NO., under basal physiological conditions the biological activity of NO. in the blood vessels is not completely lost. It has been shown that three factors reduce the efficiency of hemoglobin to scavenge NO.: a so-called red blood cell-free zone created close to the vessel wall by intravascular flow, an undisturbed layer around the red blood cells--where the NO. concentration is much smaller than the bulk concentration--and/or the red blood cell membrane. Alternatively, it has been proposed that NO. binds to Cys beta 93 of oxyHb, is liberated after deoxygenation of Hb, and consequently allows for a more effective delivery of O2 to peripheral tissues. However, because of the extremely fast rate of the reaction between NO. and oxyHb, experiments in vitro lead to artefactual production of large amounts of S-nitroso-hemoglobin. These results, together with other data, which challenge most steps of the NO.-transporter hypothesis, are discussed. PMID- 14558474 TI - [Fatty acid composition of brain hemispheres of spontaneously hypertensive rats suckled by female Wistar rats]. AB - Total lipid fatty acid composition was investigated in brain hemispheres of male Spontaneously Hypertensive Rats (SHR), compared with normotensive Wistar Kyoto rats (WKY) used as controls. Both strains were suckled by adoptive Wistar mothers, and then fed a standard diet after weaning. No difference was observed between the two hemispheres of WKY killed either at 10 or 30 days. In SHR killed at 10 days, the two hemispheres showed differences, SHR left hemispheres exhibiting greater fatty acid composition changes than those of WKY, phenomenon that toned down at 30 days. Hence, SHR pups showed a different total lipid fatty acid composition of their brain hemispheres when compared with their WKY controls, though the two strains received the same diet. Genetically programmed hypertension might be, directly or not, involved in these changes. PMID- 14558475 TI - Gravitational stress on germinating Pinus pinea seeds. AB - In the germination of lipid-rich seeds, the glyoxylate cycle plays a control role in that, bypassing the two decarboxylative steps of the Krebs cycle; it allows the net synthesis of carbohydrates from lipids. The activity of isocitrate lyase, the key enzyme of the glyoxylate cycle, is an indicator of the state of seed germination: stage of germination, growth of embryo, activation and progress of protein synthesis, depletion of lipidic supplies. In order to investigate the effects of gravity on seed germination, we carried out a study on the time pattern of germination of Pinus pinea seeds that were subjected to a hypergravitational stress (1000 g for 64 h at 4 degrees C), either in a dry or in a wet environment, before to be placed in germination plates. During the whole time of germination, we monitored the state of embryo growth and the most representative enzymes of the main metabolic pathways. In treated wet seeds, we observed an average germination of only 20% with a slowdown of the enzyme activities assayed and a noticeable degradation of lipidic reserves with respect to the controls. These differences in germination are not found for dry seeds. PMID- 14558476 TI - The geographical segregation of human lice preceded that of Pediculus humanus capitis and Pediculus humanus humanus. AB - In order to investigate human-louse phylogeny, we partially sequenced two nuclear (18S rRNA and EF-1 alpha) and one mitochondrial (COI) genes from 155 Pediculus from different geographical origins. The phylogenetic analysis of 18S rRNA and EF 1 alpha sequences showed that human lice were classified into lice from Sub Saharan Africa and lice from other areas. In both clusters, head and body lice were clearly grouped into two separate clusters. Our results indicate that the earliest divergence within human pediculidae occurred between African lice and other lice, and the divergence between head and body lice was not the result from a single event. PMID- 14558478 TI - Video-logging for examining biogenic structures in deep heterogeneous subsurface sediments. AB - Video-logging through a transparent tube is described as an alternative method for examining in situ the distribution of biogenic structures within the deep layers of heterogeneous sediments in which undisturbed cores are hard to be obtained. The method comprises four successive phases: the installation of transparent tubes into the sediment, the lowering of a video-camera for scanning sediments through the tube walls, the numerical treatment of images for correcting deformations and obtaining panoramic views of discrete sediment layers, and the mapping of biogenic structures. The method was used successfully to determine the density and depth distribution of galleries made by the oligochaetae Tubifex tubifex (Muller, 1874) within the infiltration bed of a stormwater basin. PMID- 14558477 TI - Conservation and control strategies for the wolf (Canis lupus) in western Europe based on demographic models. AB - Securing the long-term acceptance of large carnivores such as the wolf (Canis lupus) in Europe and North America raises a difficult challenge to conservation biologists: planning removals to reduce depredations on livestock while ensuring population viability. We use stochastic-stage-structured population models to investigate wolf population dynamics and to assess alternative management strategies. Among the various management strategies advocated by agencies, zoning that involves eliminating wolves outside a restricted area should be designed with caution, because probabilities of extinction are extremely sensitive to the maximum number of packs that a zone can support and to slight changes in stage specific survival probabilities. In a zoned population, viability is enhanced more by decreasing mortality rates in all classes than by increasing wolf zone size. An alternative to zoning is adaptive management, where there is no limit on pack number but population control can be operated whenever some predefined demographic conditions are met. It turns out that an adaptive management strategy that removes a moderate percentage (10%) of the population following each year of more than 5% of total population growth would provide visible actions addressing public concerns while keeping extinction probability low. PMID- 14558479 TI - Phylogenetic relationships within the fern genus Hymenophyllum s.l. (Hymenophyllaceae, Filicopsida): contribution of morphology and cytology. AB - The phylogenetic relationships of Hymenophyllum and its segregate genera Cardiomanes, Hymenoglossum, Rosenstockia, Serpyllopsis and Microtrichomanes are addressed, using 31 morphological characters of the sporophyte and one cytological character. As expected, this study reveals considerable morphological heterogeneity within the genus sensu lato, but several apomorphic changes allow support for some clades. Four unresolved taxa, Cardiomanes, Hymenoglossum, Diplophyllum and Mecodium pro parte are probably the most basal elements in Hymenophyllum. The analysis also suggests the polyphyly of Mecodium, and two unexpected associations: Sphaerocionium together with Microtrichomanes; and a broad clade composed of subg. Hymenophyllum, Hemicyatheon and Craspedophyllum, genera Rosenstockia and Serpyllopsis, and subsect. Leptocionium and Amphipterum. These associations appear justified by morphological, cytological or geographical data, and most of them are in agreement with preliminary molecular results. PMID- 14558480 TI - [Apoptosis: physiological cell death and its role in pathogenesis of diseases]. AB - Apoptosis is a physiological process of cell death by which a single cell may be eliminated from the living tissue. Since the process is mediated by specific proteins encoded in the host's genome, it is also a programmed cell death. Apoptosis is responsible for tissue remodelling during the development and turnover of normal tissue (e.g. haematopoietic cells) throughout the life span of multicellular organisms. In contrast to cells undergoing a pathological cell death (necrosis), the morphological changes that accompany apoptosis are characterised by condensation of chromatin and cytoplasm and subsequent fragmentation of the cell into small membrane-bound segments called apoptotic bodies. The maintenance of membrane integrity in apoptosis prevents the release of deleterious cytoplasmic substances and the activation of inflammatory responses. Apoptosis can be initiated by a variety of events arising either within the cell (p53) or externally (death receptor ligands engaging specific cell surface receptors of the target cell). Following recognition of the stimuli and clustering of membrane proteins into a death domain, certain cytoplasmic proenzymes are converted to their active form (caspases). After the death effector machinery is activated, the cell enters the irreversible common degradation phase of cytoskeletal disorganisation. Survival proteins (bcl-2 family) control the caspase-driven engine of destruction. Disordered apoptotic process participates in the pathogenesis of various diseases, such as neoplasms, chronic inflammatory or systemic autoimmune diseases, and other conditions. PMID- 14558481 TI - [Does diabetes mellitus affect the course and prognosis of ischemic stroke?]. AB - Although diabetes is a well-known risk factor for ischemic stroke, its role in ischemic stroke outcome has not been clarified yet. Stroke subtypes according to the TOAST (Trial of Org 10172 in Acute Stroke Treatment) classification, history of hypertension, serum glucose levels, blood pressure and OCSP (Oxfordshire Community Stroke Project) clinical types of admission, the presence of infections and seizures in the acute phase of illness, duration of hospitalisation, early and in-hospital mortality in diabetics and non-diabetic stroke patients were studied. CT scans in both groups were analysed by the size, localisation and number of ischemic foci. Significant differences were found only as regards the history of hypertension, as well as glucose levels and blood pressure on admission. The incidence of arterial hypertension prior to ischemic stroke was higher in the diabetic group. These patients had significantly higher blood glucose, systolic and diastolic blood pressure level on admission than had the non-diabetic group. No differences were found between the two groups on any other analysed variables. Our observations suggest that diabetes has no effect on the course and outcome of ischemic stroke. PMID- 14558482 TI - [Cerebrovascular reactivity in patients with mild head injury]. AB - Transcranial Doppler sonography (TCD) is a non-invasive method that can be repeated to measure blood flow velocity in intracranial arteries and to assess cerebrovascular reactivity in patients with a variety of neurological diseases. The aim of this study was Doppler sonography evaluation of blood flow and reactivity in the middle cerebral artery in patients at different stages after mild head injuries. The dynamic assessment of blood flow velocity and pulsatility index both at rest and after hyperventilation in 73 patients with mild head injuries was carried out on the day of injury, the third day, sixth day and after 6 months to 5 years. The control group consisted of 61 healthy volunteers. Obtained results indicate an increase in blood flow velocity in the middle cerebral artery on the day of injury and a decrease within the following days in younger patients (aged xleq 30 yrs). Also their cerebrovascular reactivity was significantly lower. In older persons (age > 30 yrs) and in patients with remotely suffered injuries, neither blood flow velocity in the middle cerebral arteries nor cerebrovascular reactivity differed significantly from the results of the respective age control groups. The findings justify a conclusion that mild head injury is followed by changes both in the blood flow and cerebrovascular reactivity. In younger patients with mild head injuries these haemodynamic disturbances seem to be connected with altered activity of the autonomic system. PMID- 14558483 TI - [Depression in patients with Parkinson's disease]. AB - The aim of our study was to assess the frequency of depression in group of patients with Parkinson's disease (PD) who fulfilled the diagnostic criteria of PD, had normal CT scans and responded well to L-dopa treatment. The sample consisted of 73 consecutive patients (34 women and 39 men), mean age 65.7 (41-81) years, mean duration of disease 6.7 years. Besides neurological examination, in all the patients the degree of motor impairment was evaluated using the UPDRS, H Y, and SE scales. Moreover, a sociodemographic questionnaire, psychological tests (MADRS, MMSE), and a quality of life scale (PDQ-39) were used. Depression (MADRS scores > 19) was found in 25 (34.2%) of the patients, with major depression (scores > 28) diagnosed in 7 patients (9.5%) and moderate depression (scores between 20 and 28)--in 18 cases (24.6%). In comparison to non-depressed patients, those with depression were older by 0.9 years on the average, their onset of the disease occurred later by 1.7 years, and their mean duration of the disease was longer by 2.6 years. These differences were not statistically significant. Dementia (MMSE scores < or = 23) did not differentiate between the two groups: it was found in 27 depressed patients (37.4%) and in 26 (35.6%) of those without depression. Patients in the depressed group suffered statistically more often from sleep disorders (19 vs. 14; p < 0.001). In this group motor impairment was significantly more marked, as measured by the UPDRS (32.2 vs. 46.8; p < 0.001) and H-Y (2.54 vs 2.98; p < 0.007), and their quality of life as measured by PDQ 39 questionnaire was significantly lower (36.4 vs. 82.24; p < 0.00002). Our data indicate the presence of depression in 34.2% of the sample, i.e. a somewhat lower prevalence rate than that reported in other studies. This may be due to the fact that only outpatient population was analysed, and outpatients are seldom categorized as degree 4 and 5 on the H-Y scale. Depression on PD patients was correlated with their more severe motor disability and considerably lower quality of life. This may suggest a relationship with progression of the disease and more pronounced changes in cerebral neurotransmitters (i.e. endogenous origin), or PD patient's response to their limited mobility and isolation in later stages of the disease (i.e. reactive origin). However, the two factors--endogenous and reactive -may be overlapping, since a majority of PD patients suffer from mild to moderate depression. PMID- 14558484 TI - [Changes in head circumference and ventricular system size after neuro-endoscopic third ventriculostomy in children]. AB - The aim of the study was to compare changes in the head circumference ventricular system size after neuroendoscopic third ventriculostomy with those following shunt implantation in children suffering from chronic hydrocephalus. The data were analysed to establish criteria of success of neuroendoscopic procedures. In the years 1999-2001 neuroendoscopic third ventriculostomy was performed in 59 children at the Neurosurgery Department of the Research Institute of Polish Mothers' Memorial Hospital. However, the sample analysed in the paper consists of 29 children (16 boys, 13 girls aged from 18 days to 18 years, mean age 7.03, SD = 7.11 years) with chronic hydrocephalus successfully treated with neuroendoscopic procedures. The control group consists of 59 children (31 boys, 28 girls) selected out of 80 patients who underwent primary shunt implantation at the same Neurosurgical Department in the years 1992-1994. The control children (aged from 2 weeks to 9 months, mean age 2 months, SD = 1.92 months) did not need shunt revision during the clinical observation period. The ventricular system size was assessed in terms of the Frontal Index, while postoperative changes in the system size were expressed by the ratio of the Final Frontal Index to the Baseline Frontal Index. If the ventricular system size remained the same, the ratio was 1; if its size decreased after surgery, the ratio was less than 1, while any increases in the system size were reflected by a ratio over 1. Moreover, the head circumference (HC) was measured before and after surgery only in infants and neonates with non-communicating hydrocephalus. HC was expressed in centiles using the centile chart developed by Kurniewicz-Witczakowa for various age and sex groups of Polish children. The analysis included also post-surgery changes in HC over the observation period, in terms of the difference between the baseline HC value and HC measurements in relation to the observation period duration. A positive sign of this index evidenced a decrease in the rate of HC enlargement, while a negative sign--an increased rate of HC growth. The mean HC at the end of the observation period was 72.96 centile in the neuroendoscopy group and 52.36 centile in children after shunt implantation. The reduction of head circumference following neuroendoscopic procedures was significantly smaller than that after shunt implantation, as the average decrease in HC after neuroendoscopy was only 0.4 centile as compared to about 18 centiles after shunt implantation. In the neuroendoscopy group a relationship was found between HC and age: in newborns HC was significantly smaller than that in infants (20.25 and 82.55 centiles, respectively). An analysis of HC changes (in centiles) in relation to the time since the surgery in all the children aged under 1 year, successfully treated with neuroendoscopic procedures, indicated no tendency to a steady increase in the rate of HC enlargement, even though in many cases the HC after surgery was larger than that prior to the surgery. As regards changes in the ventricular system size, the average ratio of Final to Baseline Frontal Index was 0.9 in the neuroendoscopy group and 0.5 in the group after shunt implantation. The ventricular system turned out to be significantly larger in infants after neuroendoscopy than in other age groups (the mean Frontal Index values were 0.65 vs. 0.53, respectively). No tendency to constant enlargement of the ventricular system size after neuroendoscopy was found. In children with non-communicating hydrocephalus due to Chiarii II malformation a mild enlargement of the ventricular system was seen after successful neuroscopy (the ratio of the Final to Baseline Frontal Index amounted to 1.3). CONCLUSIONS: The rate of head circumference (HC) enlargement in infants after succeeded neuroendoscopic procedures did not continually increase during the postoperative period, although their HC expressed in centiles could be higher than that before surgery. The average reduction of the ventricular system size was much smaller after neuroendoscopic ventriculostomy than than after shunt implantations. In children with Chiarii II malformation and in infants the ventricle system size may be somewhat increased in comparison to pre-operative levels. However, no tendency to a steady enlargement with time was found either in the HC or in the ventricle system size. PMID- 14558485 TI - [Does left-handedness affect the pattern of cerebral blood flow during cognitive activity?]. AB - In most people the left hemisphere of the brain is dominant for language functions. Because of an increased incidence of atypical language lateralisation in the right hemisphere of left-handed neurological patients, more systematic studies on handedness and cerebral asymmetry of language have been undertaken. The present study is aimed at clarification of the relationship between handedness and language dominance in healthy subjects. Lateralisation was measured directly using functional transcranial Doppler sonography in left-handed subjects. Twenty-six individuals participated in the study. Three kinds of tasks were used, differing in the material involved and in appropriate strategies to be employed by the subjects. Two important parameters of the MCA blood flow were analysed: mean relative increase in the blood flow velocity (MDV) and specific patterns of cognitive task performance (i.e. performance profiles). Our results indicate that the incidence of the right hemisphere dominance for language depends on the degree of handedness, since only in the group of consistent left handers language was right lateralised. In other left-handers both cerebral hemispheres were functionally equivalent. The measurement of MCA blood flow velocity changes using transcranial Doppler ultrasonography seems to be a simple and non-invasive method of assessing the functioning of the two hemispheres of human brain. PMID- 14558486 TI - [Cerebral amyloid angiopathy as a cause of strokes]. AB - Cerebral amyloid angiopathy (CAA), a condition affecting the elderly in a way similar to that of Alzheimer's disease, results from amyloid deposition within small and medium arteries of the cerebral leptomeninges and cerebral cortex. Next to atheromatosis, amyloidosis is the second most frequent cause of cerebral haemorrhage, especially recurrent. The most recent publications suggest that amyloidosis may also cause transient ischaemic attacks (TIA), cerebral infarcts, Binswanger's type leukoencephalopathy, symptoms resembling these of cerebral pseudotumour, and other dysfunctions. A definite diagnosis of cerebral amyloid angiopathy may be determined by autopsy, and sometimes intravital cerebral biopsy is performed. A clinical diagnosis of probable CAA is based on the presence of multiple superficial haemorrhages in the elderly people without hypertension. No CAA treatment methods are known yet. However, to prevent haemorrhages in CAA it is important that anticoagulants and antiplatelet drugs should be avoided in the treatment of cerebral and other ischaemic disorders, because of an increased risk for haemorrhage. PMID- 14558488 TI - [Ischemic manifestation of adult moyamoya disease: a case report]. AB - Moyamoya disease is a chronic non-inflammatory cerebrovascular steno-occlusive disorder of unknown origin, with a typical pattern of collateral netlike vessels visualized in angiography. In Poland it is a relatively rare condition. A case of a 44-year-old non-Japanese woman with moyamoya disease is presented. Ischemic manifestation of the disease, rather uncommon in adults, and a long asymptomatic post-operative period are discussed. PMID- 14558487 TI - [Diagnostic difficulties in herpes simplex encephalitis patients: report of three cases]. AB - Herpes simplex encephalitis (HSE) is a rare infection of the central nervous system. The prognosis is poor, even in those receiving anti-herpetic therapy (about 20% of patients die, and in 40% of cases neurological sequelae are severe). The diagnosis is based on clinical signs of encephalitis, typical pathological changes in MRI, and on the presence of anti-HSV antibody and/or HSV DNA in the cerebrospinal fluid. Three cases of encephalitis are presented. The clinical course and pathological changes in the temporal lobes seen in MRI were characteristic of the HSV etiology. Early intrathecal synthesis of anty-HSV antibodies were not detected despite an increase in their serum level in the course of the disease. HSV-DNA detection with PCR was not performed. Nevertheless in all cases aciclovir therapy was introduced on the first day of hospitalization. Possibilities of diagnosing HSE on the basis of clinical signs and MRI scans during the first week of the disease are discussed, as well as the tissue whether the course of treatment would be changed accordingly to the PCR test results. PMID- 14558489 TI - [Familial congenital bilateral Duane's retraction syndrome (Stilling-Turk-Duane syndrome). A case report]. AB - The authors describe a case of Duane's Retraction Syndrome, which is an unfrequent cause of eye movement disturbances. The classic syndrome is characterized by a disability for horizontal eyes movements without concurrent diplopia. It is a multi-etiological, congenital or acquired syndrome. PMID- 14558490 TI - [Deep brain stimulation of the Vim nucleus of the thalamus in the treatment of parkinsonian tremor]. AB - Deep brain stimulation (DBS) of the ventral intermediate thalamic nucleus (Vim) has been recently introduced by Benabid and his colleagues as a new surgical procedure in the treatment of tremor-dominant Parkinson's disease (PD). The advantage of DBS Vim over lesioning (thalamotomy) is its reversibility and adjustability with the same clinical effect, but without the need to make a destructive thalamic lesion. In this procedure high-frequency stimulation is employed to simulate a thalamic lesion using an implanted electrode connected to a subcutaneously placed neuropacemaker. Four patients with tremor-dominant PD were included in the study. There were 3 men and one women. Three stimulators were implanted in the left and one in the right cerebral hemisphere. The patients were evaluated using clinical scales, before and up to 24 months after surgery. Adverse effects associated with chronic Vim stimulation were mild and reversible. Chronic thalamic stimulation is effective for drug-resistance parkinsonian tremor suppression, with few adverse side-effects. The method results in a significant improvement of function. PMID- 14558491 TI - [Bilateral deep brain stimulation of subthalamic nucleus STN in the surgical treatment of Parkinson's disease]. AB - Dopamine deficiency in the nigrostriatal system leads to a series of changes in the basal ganglia, resulting in an increased neuronal activity of the subthalamic nucleus (STN). Reduction of the STN glutaminergic excitatory effect on the main output structures of the basal ganglia (globus pallidum pars interna GPi and substantia nigra pars reticulata SNr) is accompanied by a marked alleviation of parkinsonian motor sings in the MPTP monkey model of parkinsonism. Also a high frequency stimulation of STN in the MPTP monkey model of parkinsonism produced the same clinical effect as did lesioning. Due to these observations bilateral deep subthalamic stimulation was introduced in the treatment of PD patients with severe akinetic-rigid form of this disease. Four patients with akinetic-rigid PD form of PD were included in the study. The electrodes for deep brain stimulation were implanted in two separate surgical interventions in every case. The second implantation was performed not earlier than at least 3 months after the first procedure. Evaluations using the UPDRS were conducted before surgery in "on" and "off" conditions and at 3, 6 and 12 months after the bilateral implantation. Bilateral DBS STN seems to be the best stereotactic target in controlling motor symptoms in the "off" condition in the treatment of PD patients with severe symptoms. The technique enables a dramatic reduction in the daily dose of L-dopa. PMID- 14558492 TI - [Hemifacial spasm as an early symptom of posterior fossa meningioma]. AB - A rare case of a 47-year old female with hemifacial spasm of the one-year duration as the initial and only symptom of posterior fossa meningioma is reported. At first the patient did not consent to surgery and was unsuccessfully treated with anticonvulsants. A month prior to her admission she started to complain of headache and balance disorder, and developed sensory loss to light touch in II and III branches of the right V nerve. CT revealed a huge posterior fossa meningioma. After a total removal of meningioma, the patient's hemifacial spasm disappeared. PMID- 14558494 TI - [Report on the course "Microsurgical procedure planning", Mainz, Sept. 27-30, 2002]. PMID- 14558493 TI - [Own experience in surgical treatment of the pineal region and midbrain tumors via the infratentorial approach]. AB - Nine cases of tumours located in the pineal and midbrain region in adults operated on between November 1998 and July 2002 in Dept. of Neurosurgery, Central Military Hospital in Warsaw are reported. The patients (2 men and 7 women) were aged from 27 and 69 years (mean age 43.6 years). Their main initial symptoms were caused by hydrocephalus. The histopathological examination revealed anaplastic pinealoma in 2 cases, and pineocytoma, pineal cyst, mesencephalic glial cyst, protoplasmatic astrocytoma, epidermoid cyst, unclear glial scar, and papillary ependymoma in single cases. Five patients had been treated, usually elsewhere, with shunt implantation prior to the surgery. Occipito-suboccipital osteoplastic craniotomy was performed in each case and tumours were totally removed microsurgically via the infratentorial epicerebellar approach. In one case a part of the glial periaqueductal tumour was resected additionally via the fourth ventricle and aqueduct in one stage. Postoperative haematomas in the third and fourth ventricle were found in 2 cases. Main complaints after the surgery included transient diplopia. All the patients improved significantly and resumed their previous life activities. Follow-up ranged from 3 to 44 months. Three patients with pineal tumours and one with a small postoperative ependymoma recurrence were irradiated after the surgery. One patient had been irradiated prior to surgery. Very good results of the surgical treatment of tumours in this area suggest that such patients should be referred earlier to one stage surgical management, as the procedure is easier to perform and shunt implantation may be avoided. PMID- 14558495 TI - [Report on the 7th Congress of the EMN in Newcastle, June 2002]. PMID- 14558496 TI - Facial pain as first manifestation of lung cancer: a case of lung cancer-related cluster headache and a review of the literature. AB - Facial pain can, on rare occasions, be the presenting symptom of lung cancer. This report describes a patient with non-metastatic lung cancer, which was associated with attacks of debilitating facial pain, presenting as cluster headache. Moreover, 32 reported cases of lung cancer-related facial pain (including the present one) are reviewed, and their clinical features are summarized. The facial pain is almost always unilateral, and is most commonly localized to the ear, the jaws, and the temporal region. The pain is frequently described as severe and aching, and may be continuous or intermittent. Aggravation and expansion of the pain, digital clubbing, increased erythrocyte sedimentation rate, and hypertrophic osteopathy, may contribute to the diagnosis. Referred pain, due to invasion or compression of the vagus nerve, as well as paraneoplastic syndrome secondary to the production of circulating humoral factors by the malignant tumor cells, is implicated in the pathophysiology of facial pain associated with non-metastatic lung cancer. Radiotherapy and tumor resection with vagotomy are very effective in aborting the facial pain. Thus, lung cancer should be included in the differential diagnosis of facial pain that is atypical and/or refractory to treatment. PMID- 14558497 TI - Short-pulse generation at 10 microm in an active cw-injected ring laser cavity. AB - Continuous coherent light conversion in a train of short pulses with good efficiency is possible with a multipass interferometer in which the frequency is shifted at every pass with an acousto-optic frequency shifter. This technique allows one to generate a spectrum made of equidistant components, interferences of which build intense light pulses. Unfortunately, both the width and efficiency of the pulses are limited by the losses undergone by the waves traveling through the interferometer cavity. Improvement of the pulse duration, the peak intensity, and the contrast can be expected in such an experiment when an amplifier is set up inside the cavity. I report on theoretical computations related to this apparatus and apply this theoretical model to a high-pressure CO2 amplifier. PMID- 14558498 TI - [Results of hallux valgus therapy after initial surgical correction]. AB - Authors present own experience after use surgical correction technique in order to heal the complex hallux valgus deformity. In period 1999-2002 we used this technique on 5 patients (4 females, 1 male) with 6 surgical procedures (1 female had bilateral treatment). Indications for surgical treatment were functional problems and x-ray angle between the first and second metatarsals as well as valgus angle of the first metatarsophalangeal joint which were much higher. Pre surgical, possibility of correction angle was exact by x-ray planing. The same examination was done after surgical treatment. After surgical care and rehabilitation program all patients had well biomechanical foot relation and they had not previous functional problems. PMID- 14558499 TI - Pharmacological evaluation of 5-[2-[4-(2-methoxy-phenyl)-piperazin-1-yl]-ethyl] 1,3-dihydro-benzimidazole-2-thione as a potential atypical antipsychotic agent. PMID- 14558500 TI - [Dutch Dentistry Association Congress: dealing with knowledge]. PMID- 14558501 TI - Tick-borne relapsing fever under-reported. PMID- 14558502 TI - AIDS foundation seeks withdrawal of approval for HIV drug combination. PMID- 14558503 TI - Retroviruses may trick their way around the human body. PMID- 14558504 TI - Time to prioritise tuberculosis laboratory services. PMID- 14558505 TI - Delays hinder outbreak investigations in India. PMID- 14558506 TI - Sweet solution to superbug infections? PMID- 14558507 TI - The five stages of asthma. PMID- 14558508 TI - Bring on the Brahms. PMID- 14558509 TI - Prospects fade for drug benefit for US elderly. Costly legislation appears dead in face of record deficit and grim economic news. PMID- 14558510 TI - Experimental treatment may show promise for vCJD. PMID- 14558511 TI - Narcolepsy drug could be approved for wider use. PMID- 14558512 TI - "Well, they were doing it too.". PMID- 14558513 TI - Abstracts of the International Association of Biomedical Gerontology 10th Congress. 19-23 September 2003, Cambridge, United Kingdom. PMID- 14558514 TI - EUROSPINE 2003. Abstracts of the 5th annual meeting of the Spine Society of Europe. September 30-October 4, 2003. Prague, Czech Republic. PMID- 14558515 TI - Brain Research cumulative author and subject indexes 2002, volumes 924-958. PMID- 14558516 TI - Abstracts of the 17th annual North American Cystic Fibrosis Conference. Anaheim, California, USA. October 16-19, 2003. PMID- 14558517 TI - Abstracts of the 10th biennial meeting of the European Behavioural Pharmacology Society. Antwerp, Belgium, 6-9 September 2003. PMID- 14558518 TI - Abstracts of the 25th Congress of the European Society of Parenteral and Enteral Nutrition. Cannes, France, 20-23 September 2003. PMID- 14558519 TI - Abstracts of the British Pharmaceutical Conference. Harrogate, United Kingdom, September 15-17, 2003. PMID- 14558520 TI - [The 31st annual meeting of the Japanese Society of Clinical Immunology. Tokyo, Japan. October 9-10, 2003. Abstracts]. PMID- 14558522 TI - Assessing the future hydrogen economy. PMID- 14558521 TI - [The 47th annual meeting of the Japanese Society for Medical Mycology. Tokyo, Japan. October 16-17, 2003. Program and Abstracts]. PMID- 14558523 TI - Assessing the future hydrogen economy. PMID- 14558524 TI - [Early-stage conservative removal of scars and delayed skin grafting for severe burns in the neck: report of 13 cases]. AB - OBJECTIVE: To optimize the treatments for severe burns in the neck to prevent the hypertrophy, contraction and deformation of the scars and avoid repeated surgical repair. METHODS: At the early stage of severe neck burns, conservative removal of the scabs was adopted with a small portion of the necrotic tissues preserved. Wound addressing was performed with anti-infection measures supplemented with the application of agents for dissolving the scars, promoting the epithelial growth and preventing local inflammation. Wound repair with large pieces of autologous skin grafts of medium or full thickness was performed with reasonable delay, after which the neck was fixed with thermoplastic splints. RESULTS: The wounds of 13 patients in this survey healed smoothly within 3 weeks, and no obvious hypertrophy, contracture or deformation of the scars was observed. The patients complained no restrained movement of the neck, and adhesion did not occur in the chin, neck or the chest. CONCLUSION: The method described above may help promote the healing of the neck burns and reduce the incidences of hypertrophy, contracture and deformation of the scars, which avoids further operations for the repair. PMID- 14558525 TI - Plant scents--mediators of inter-and intraorganismic communication. PMID- 14558526 TI - Apolipoprotein E (ApoE) and ApoE-rich high-density cholesterol as potential early diagnostic markers of congenital biliary tract anomalies. PMID- 14558527 TI - ALL and fractures. PMID- 14558528 TI - Prospective study of carboplatin-based chemotherapy for pediatric germ cell tumors. PMID- 14558529 TI - Phase II study of paclitaxel plus gemcitabine in refractory germ cell tumors (E9897): a trial of the Eastern Cooperative Oncology Group. PMID- 14558530 TI - Ototoxicity: understanding oxidative mechanisms. PMID- 14558531 TI - Abstracts of the North American Congress of Clinical Toxicology Annual Meeting. September 4-9, 2003. Chicago, Illinois, USA. PMID- 14558532 TI - Management preferences following radical inguinal orchidectomy for Stage 1 testicular seminoma in Australasia. PMID- 14558533 TI - Late recurrence in 1263 men with testicular germ cell tumors. Multivariate analysis of risk factors and implications for management. PMID- 14558534 TI - Role of P53 and MDM2 in treatment response of human germ cell tumors. PMID- 14558535 TI - Bilateral testicular germ cell tumors: twenty-year experience at M. D. Anderson Cancer Center. PMID- 14558536 TI - Prevalence of renal cell carcinoma in patients with ESRD pre-transplantation: a pathologic analysis. PMID- 14558537 TI - Emphysematous pyelitis presenting as an acute abdomen in an end-stage renal disease patient treated with peritoneal dialysis. PMID- 14558539 TI - Adrenal incidentaloma--experience of a standardized diagnostic programme in the Swedish prospective study. PMID- 14558538 TI - A low-dose adrenocorticotropin test reveals impaired adrenal function in cancer patients receiving megestrol acetate therapy. PMID- 14558541 TI - Association between competing interests and authors' conclusions: epidemiological study of randomised clinical trials published in the BMJ. PMID- 14558544 TI - Endocrine sequelae of cancer therapy in childhood. PMID- 14558546 TI - Should measurement of maximum urinary flow rate and residual urine volume be a part of a "minimal care" assessment programme in female incontinence? PMID- 14558545 TI - Antimicrobial prophylaxis for urinary tract infection in persons with spinal cord dysfunction. PMID- 14558547 TI - Urinary incontinence in Canada. National survey of family physicians' knowledge, attitudes, and practices. PMID- 14558548 TI - Quality of life and seeking help in women with urinary incontinence. A population based study. PMID- 14558550 TI - Tension-free vaginal tape operation: results of the Austrian registry. PMID- 14558549 TI - Review of cadaveric allografts in urology. PMID- 14558551 TI - The Second Intercontinental Landfill Research Symposium. PMID- 14558552 TI - Vaginal palpation of pelvic floor muscle strength: inter-test reproducibility and comparison between palpation and vaginal squeeze pressure. PMID- 14558553 TI - Prevention of postpartum stress incontinence in primigravidae with increased bladder neck mobility: a randomised controlled trial of antenatal pelvic floor exercises. PMID- 14558554 TI - Posterior tibial nerve stimulation in the treatment of urge incontinence. PMID- 14558555 TI - Efficacy of botulinum-A toxin in children with detrusor hyperreflexia due to myelomeningocele: preliminary results. PMID- 14558556 TI - Differing effects of N(G)-monomethyl L-arginine and 7-nitroindazole on detrusor activity. PMID- 14558557 TI - Bladder injection of "naked" hSlo/pcDNA3 ameliorates detrusor hyperactivity in obstructed rats in vivo. PMID- 14558558 TI - Anterior vaginal repair for urinary incontinence in women. PMID- 14558559 TI - Conservative management for post prostatectomy urinary incontinence. PMID- 14558560 TI - Use of heparin and protamine sulfate during laparoscopic donor nephrectomy. PMID- 14558561 TI - Donor characteristics associated with reduced graft survival: an approach to expanding the pool of kidney donors. PMID- 14558562 TI - Exercise training improves cardiopulmonary function and quality of life in postmenopausal breast cancer survivors. PMID- 14558563 TI - Kidney allograft and patient survival in type I diabetic recipients of cadaveric kidney alone versus simultaneous pancreas kidney transplants: a multivariate analysis of the UNOS database. PMID- 14558564 TI - Controlled endurance or strength training of the neck muscles decreases pain and disability in women with chronic neck pain. PMID- 14558565 TI - Parental education does not reduce morbidity in pre-school children with asthma. PMID- 14558566 TI - Prolongation of long-term kidney graft survival by a simultaneous liver transplant: the liver does it, and the heart does it too. PMID- 14558567 TI - An interdisciplinary and multifactorial prevention program reduces falls in older people in residential care. PMID- 14558568 TI - Expression of TGF-beta and fibrogenic genes in transplant recipients with tacrolimus and cyclosporine nephrotoxicity. PMID- 14558569 TI - Morning blood pressure surge and the risk of stroke. PMID- 14558570 TI - Morning blood pressure surge and the risk of stroke. PMID- 14558571 TI - [Neonatal cholestasis]. PMID- 14558572 TI - [Hepatic candidiasis in patients with acute myelogenous leukemia]. PMID- 14558573 TI - [Ultrastructure of chronic liver diseases - mitochondria and mitochondrial hepatopathies]. PMID- 14558574 TI - Bibliography. Current world literature. Imaging and echocardiography. PMID- 14558575 TI - Bibliography. Current world literature. Prevention. PMID- 14558576 TI - The Fresno Sanitary Landfill in an American cultural context. PMID- 14558577 TI - September 11 and the mourning after: reflections on collecting and interpreting the history of tragedy. PMID- 14558578 TI - The Holocaust on film: the feature films. PMID- 14558579 TI - West Germany and unified German cinema's difficult encounter with the Holocaust. PMID- 14558580 TI - Holocaust iconography in American feature films about Neo-Nazis. PMID- 14558581 TI - Does it really matter who places dental implants? PMID- 14558582 TI - Theodor Kotulla's excerpts from a german life. PMID- 14558583 TI - All rules barred: a defense of Spielberg's Schindler's list. PMID- 14558584 TI - M. du Bois-Reymond goes to Paris. AB - This article examines the science of electrophysiology developed by Emil du Bois Reymond in Berlin in the 1840s. In it I recount his major findings, the most significant being his proof of the electrical nature of nerve signals. Du Bois Reymond also went on to detect this same 'negative variation', or action current, in live human subjects. In 1850 he travelled to Paris to defend this startling claim. The essay concludes with a discussion of why his demonstration failed to convince his hosts at the French Academy of Sciences. PMID- 14558585 TI - Parental evaluation of quality of life measures following pediatric dental treatment using general anesthesia. AB - The purpose of this study was to examine (a) parental satisfaction with the dental care their child received under general anesthesia, and (b) perception of the impact of this care on physical and social quality of life. The sample included 45 children (median age 50 months, 26 boys and 19 girls). Data were collected using a 1-page survey instrument completed by the parent at the first follow-up appointment. Dichotomous dependent variables were developed to measure parental satisfaction, dental outcome, and social impact of treatment. There was an overwhelmingly positive impression with dental outcomes (pain relief and improved masticatory efficiency). Parental perceptions in the social dimension were also positive. Parents reported more smiling, improved school performance, and increased social interaction. Relative to overall health, the majority of parents reported an improvement. Logit regression analysis revealed that absence of pain (P < .05) and increased social interaction (P < .01) had a significant impact on parents' perception of overall health. Our findings indicate that dental care under general anesthesia for preschool children has a high degree of acceptance by parents and is perceived to have a positive social impact on their child. PMID- 14558586 TI - A comparison of injection pain and postoperative pain of two intraosseous anesthetic techniques. AB - The purpose of this prospective, randomized, blinded study was to compare injection pain and postoperative pain of an apical primary X-Tip intraosseous technique to a coronal primary Stabident intraosseous technique in mandibular first molars. Using a repeated-measures design, 41 subjects randomly received 2 primary intraosseous injections at 2 separate appointments. Using a site distal to the mandibular first molar for both injections, the subjects received 1.8 mL of 2% lidocaine with 1: 100,000 epinephrine administered with the X-Tip system using an apical location in alveolar mucosa or 1.8 mL of 2% lidocaine with 1: 100,000 epinephrine administered with the Stabident system using a coronal location in attached gingiva. The pain of infiltration, perforation, needle insertion, solution deposition, mock or actual guide sleeve removal and postoperative pain were recorded on a Heft-Parker visual analogue scale (VAS) scale for the 2 intraosseous systems. The results demonstrated that the apical primary X-Tip intraosseous technique was not statistically different (P > .05) from the coronal primary Stabident technique regarding pain ratings of infiltration, perforation, needle insertion, solution deposition, mock or actual guide sleeve removal and postoperative pain (at the time subjective anesthesia wore off). However, on postoperative days 1 through 3, significantly (P < .05) more males experienced postoperative pain with the X-Tip system than with the Stabident system. PMID- 14558587 TI - An evaluation of analgesic efficacy and clinical acceptability of intravenous tramadol as an adjunct to propofol sedation for third molar surgery. AB - This article details a double-blind, randomized, placebo-controlled pilot study evaluating the analgesic efficacy and clinical acceptability of intravenous tramadol in patients undergoing surgical removal of an impacted third molar tooth under local anesthesia and intravenous sedation with propofol. Forty-five ASA status 1 dental outpatients were randomly allocated to 2 groups of 22 (group A) and 23 (group B) patients each (n = 45). Group A (T/P) received intravenous tramadol 1.5 mg/kg injected over 2 minutes, followed by a bolus dose of intravenous propofol 0.4 mg/ kg. Maintenance consisted of a continuous infusion of propofol 3 mg/kg/h, with an additional bolus dose of 0.4 mg/kg intravenously 2 3 minutes prior to the infiltration of the local anesthetic solution. Group B (P/P) patients received no tramadol but instead a saline placebo solution and an identical amount of propofol. Overall, in this study, postoperative pain was much better controlled in the group receiving tramadol 1.5 mg/kg intravenously despite there being no significant difference in the dose of propofol administered in both groups. Intravenous tramadol, when given with propofol, did not affect the cardiovascular, respiratory, and sedative effects of propofol. Following tramadol, despite being an opioid, no nausea and vomiting were reported in the early postoperative period, indicating the value of using tramadol with propofol. Thus, this pilot study demonstrated the potential use of intravenous tramadol with propofol in day-case dento-alveolar surgery. PMID- 14558589 TI - Mammal invaders on islands: impact, control and control impact. AB - The invasion of ecosystems by exotic species is currently viewed as one of the most important sources of biodiversity loss. The largest part of this loss occurs on islands, where indigenous species have often evolved in the absence of strong competition, herbivory, parasitism or predation. As a result, introduced species thrive in those optimal insular ecosystems affecting their plant food, competitors or animal prey. As islands are characterised by a high rate of endemism, the impacted populations often correspond to local subspecies or even unique species. One of the most important taxa concerning biological invasions on islands is mammals. A small number of mammal species is responsible for most of the damage to invaded insular ecosystems: rats, cats, goats, rabbits, pigs and a few others. The effect of alien invasive species may be simple or very complex, especially since a large array of invasive species, mammals and others, can be present simultaneously and interact among themselves as well as with the indigenous species. In most cases, introduced species generally have a strong impact and they often are responsible for the impoverishment of the local flora and fauna. The best response to these effects is almost always to control the alien population, either by regularly reducing their numbers, or better still, by eradicating the population as a whole from the island. Several types of methods are currently used: physical (trapping, shooting), chemical (poisoning) and biological (e.g. directed use of diseases). Each has its own set of advantages and disadvantages, depending on the mammal species targeted. The best strategy is almost always to combine several methods. Whatever the strategy used, its long term success is critically dependent on solid support from several different areas, including financial support, staff commitment, and public support, to name only a few. In many cases, the elimination of the alien invasive species is followed by a rapid and often spectacular recovery of the impacted local populations. However, in other cases, the removal of the alien is not sufficient for the damaged ecosystem to revert to its former state, and complementary actions, such as species re-introduction, are required. A third situation may be widespread: the sudden removal of the alien species may generate a further disequilibrium, resulting in further or greater damage to the ecosystem. Given the numerous and complex population interactions among island species, it is difficult to predict the outcome of the removal of key species, such as a top predator. This justifies careful pre-control study and preparation prior to initiating the eradication of an alien species, in order to avoid an ecological catastrophe. In addition, long-term monitoring ofthe post-eradication ecosystem is crucial to assess success and prevent reinvasion. PMID- 14558588 TI - Comparison of the effect of orally versus submucosally administered meperidine on the behavior of pediatric dental patients: a retrospective study. AB - The purpose of this study was to compare the effect of oral versus submucosal meperidine on the behavior of pediatric dental patients. Twenty charts (10 in each group) were retrospectively reviewed. The groups were matched for age and weight. Presedation and postsedation behavior was rated. No difference was found in the increase in cooperation between the oral and the submucosal meperidine groups. While no difference was found between the 2 groups, a larger prospective study is needed to confirm these findings. PMID- 14558590 TI - Darwinian aesthetics: sexual selection and the biology of beauty. AB - Current theoretical and empirical findings suggest that mate preferences are mainly cued on visual, vocal and chemical cues that reveal health including developmental health. Beautiful and irresistible features have evolved numerous times in plants and animals due to sexual selection, and such preferences and beauty standards provide evidence for the claim that human beauty and obsession with bodily beauty are mirrored in analogous traits and tendencies throughout the plant and animal kingdoms. Human beauty standards reflect our evolutionary distant and recent past and emphasize the role of health assessment in mate choice as reflected by analyses of the attractiveness of visual characters of the face and the body, but also of vocal and olfactory signals. Although beauty standards may vary between cultures and between times, we show in this review that the underlying selection pressures, which shaped the standards, are the same. Moreover we show that it is not the content of the standards that show evidence of convergence--it is the rules or how we construct beauty ideals that have universalities across cultures. These findings have implications for medical, social and biological sciences. PMID- 14558591 TI - Descent with modification: the unity underlying homology and homoplasy as seen through an analysis of development and evolution. AB - Homology is at the foundation of comparative studies in biology at all levels from genes to phenotypes. Homology is similarity because of common descent and ancestry, homoplasy is similarity arrived at via independent evolution. However, given that there is but one tree of life, all organisms, and therefore all features of organisms, share some degree of relationship and similarity one to another. That sharing may be similarity or even identity of structure and the sharing of a most recent common ancestor--as in the homology of the arms of humans and apes--or it may reflect some (often small) degree of similarity, such as that between the wings of insects and the wings of birds, groups whose shared ancestor lies deep within the evolutionary history of the Metazoa. It may reflect sharing of entire developmental pathways, partial sharing, or divergent pathways. This review compares features classified as homologous with the classes of features normally grouped as homoplastic, the latter being convergence, parallelism, reversals, rudiments, vestiges, and atavisms. On the one hand, developmental mechanisms may be conserved, even when a complete structure does not form (rudiments, vestiges), or when a structure appears only in some individuals (atavisms). On the other hand, different developmental mechanisms can produce similar (homologous) features. Joint examination of nearness of relationship and degree of shared development reveals a continuum within an expanded category of homology, extending from homology --> reversals --> rudiments --> vestiges --> atavisms --> parallelism, with convergence as the only class of homoplasy, an idea that turns out to be surprisingly old. This realignment provides a glimmer of a way to bridge phylogenetic and developmental approaches to homology and homoplasy, a bridge that should provide a key pillar for evolutionary developmental biology (evo-devo). It will not, and in a practical sense cannot, alter how homoplastic features are identified in phylogenetic analyses. But seeing rudiments, reversals, vestiges, atavisms and parallelism as closer to homology than to homoplasy should guide us toward searching for the common elements underlying the formation of the phenotype (what some have called the deep homology of genetic and/or cellular mechanisms), rather than discussing features in terms of shared or independent evolution. PMID- 14558592 TI - Quantitative steps in symbiogenesis and the evolution of homeostasis. AB - The merging of two independent populations of heterotrophs and autotrophs into a single population of mixotrophs has occurred frequently in evolutionary history. It is an example of a wide class of related phenomena, known as symbiogenesis. The physiological basis is almost always (reciprocal) syntrophy, where each species uses the products of the other species. Symbiogenesis can repeat itself after specialization on particular assimilatory substrates. We discuss quantitative aspects and delineate eight steps from two free-living interacting populations to a single fully integrated endosymbiotic one. The whole process of gradual interlocking of the two populations could be mimicked by incremental changes of particular parameter values. The role of products gradually changes from an ecological to a physiological one. We found conditions where the free living, epibiotic and endobiotic populations of symbionts can co-exist, as well as conditions where the endobiotic symbionts outcompete other symbionts. Our population dynamical analyses give new insights into the evolution of cellular homeostasis. We show how structural biomass with a constant chemical composition can evolve in a chemically varying environment if the parameters for the formation of products satisfy simple constraints. No additional regulation mechanisms are required for homeostasis within the context of the dynamic energy budget (DEB) theory for the uptake and use of substrates by organisms. The DEB model appears to be dosed under endosymbiosis. This means that when each free living partner follows DEB rules for substrate uptake and use, and they become engaged in an endosymbiotic relationship, a gradual transition to a single fully integrated system is possible that again follows DEB rules for substrate uptake and use. PMID- 14558593 TI - Review of the ultrastructure of the nematode body cuticle and its phylogenetic interpretation. AB - The phylogenetic interpretation of the nematode cuticle ultrastructure is reviewed within the framework of recent DNA-sequence data. In particular, the structure of the median and basal zones is discussed. Several structural elements of the cuticle seem to have arisen independently several times within the Nematoda and thus are highly homoplasious (e.g. the cortical or basal radial striae, spiral fibre layers and a fluid matrix with struts). Moreover, identifying the homology of the nematode cuticle ultrastructures is often very difficult at deep taxonomic levels. Hence, the cuticle appears to be unreliable regarding resolution of deep-level relationships in the Nematoda. However, at less inclusive taxonomic levels (e.g. families, genera, ...) the cuticle seems to be a more reliable phylogenetic marker. PMID- 14558595 TI - Local delivery of minocycline and systemic BCNU have synergistic activity in the treatment of intracranial glioma. AB - Minocycline, a tetracycline derivative, has been shown to inhibit tumor angiogenesis through inhibitory effects on matrix metalloproteinases. Previous studies have shown this agent to be effective against a rodent brain tumor model when delivered intracranially and to potentiate the efficacy of standard chemotherapeutic agents. In the present study, the in vivo efficacy of intracranial minocycline delivered by a biodegradable controlled-release polymer against rat intracranial 9L gliosarcoma was investigated to determine whether it potentiates the effects of systemic 1,3-bis (2-chloroethyl)-1-nitrosourea (BCNU). Minocycline was incorporated into the biodegradable polymer polyanhydride poly[bis(p-carboxyphenoxy)propane-sebacic acid] (pCPP:SA) at a ratio of 50:50 by weight. The release kinetics of minocycline from the polymer were assessed. For the efficacy studies, female Fischer 344 rats were implanted with 9L glioma. Treatment with minocycline delivered by the pCPP:SA polymer at the time of tumor implantation resulted in 100% survival in contrast to untreated control animals that died within 21 days. Treatment with the minocycline-polymer 5 days after tumor implantation provided only modest increases in survival. The combination of intracranial minocycline and systemic BCNU extended median survival by 82% compared to BCNU alone (p < 0.0001) and 200% compared to no treatment (p < 0.004). We conclude that local intracranial delivery of minocycline from biodegradable controlled-release polymers inhibits tumor growth and may have clinical utility when combined with a chemotherapeutic agent. PMID- 14558594 TI - Effects of thalidomide on parameters involved in angiogenesis: an in vitro study. AB - In an attempt to elucidate the mechanism(s) of action of thalidomide, a reportedly antiangiogenic molecule recently tested in the treatment of relapsing malignant gliomas, we performed an in vitro study on the following parameters: (a) effect of thalidomide on proliferation of endothelial cells; (b) effect of thalidomide on expression of alpha(v)beta3 integrin on the surface of endothelial cells; (c) effect of thalidomide on the release by endothelial cells of MMP-2, IL 8 and TNF-alpha. The results show that thalidomide inhibits endotelial cell proliferation induced by bFGF and VEGF, more so if the cells are grown on vitronectin; moreover, treatment with thalidomide reduces the release of MMP-2 and IL-8 by endothelial cells, suggesting a further pathway for the antiangiogenic activity of drug. On the other hand, thalidomide does not modify expression of alpha(v)beta3 on endothelial cells. PMID- 14558596 TI - Extracellular nucleotides and nucleosides induce proliferation and increase nucleoside transport in human glioma cell lines. AB - Extracellular purines (adenosine triphosphate (ATP), adenosine 5'-diphosphate (ADP) and adenosine) and pyrimidines (uridine 5'-triphosphate (UTP) and UDP) are important signaling molecules that mediate diverse biological effects via P1 and P2 purinergic receptors. The human glioma cell lines U87 MG, U251 MG and U138 MG were treated with purines and pyrimidines for 24 or 48 h and proliferation was measured by [3H]-thymidine incorporation, flow cytometry and cell counting. The studies showed that extracellular nucleotides and nucleosides induce proliferation of the studied glioma cells. Incorporation of [3H]-thymidine followed the order of ATP approximately equal to guanosine approximately equal to inosine approximately equal to adenosine > UTP > ADP while ATPgammaS and 2MeSATP had no effect. The effect of ATP was partially inhibited by suramin and by reactive blue 2 (RB2). Co-treatment with the following antagonists of P1 purinoreceptors DPCPX, CPT or 8PT did not block the effect of adenosine while a specific antagonist of the A3 receptor, MRS1220, totally blocked the effect of adenosine. ATP and adenosine also increased the overall uptake of [3H]-thymidine into the cell, producing a positive effect on the [3H]-thymidine incorporation measurements. These data indicate that the uptake of thymidine and proliferation of gliomas can be induced by purines and pyrimidines via both P1 and P2 purinoceptors. PMID- 14558597 TI - Differential expression of bikunin (HAI-2/PB), a proposed mediator of glioma invasion, by demethylation treatment. AB - Effective therapies for primary brain tumors continue to be elusive. Successful adjuvant therapies for CNS tumors will require a better understanding of their basic biology. Hepatocyte growth factor activator inhibitor type-2/placental bikunin (HAI-2/PB) is a serine proteinase inhibitor that has a broad inhibitory spectra against various serine proteinases. HAI-2/PB has anti-invasive effects thought to be mediated primarily by the inhibitory activity against serine proteinase-dependent matrix degradation. It has been previously demonstrated that the expression of HAI-2/PB is inversely related to degree of malignancy and possibly involved in the progression and invasion of human gliomas. Aberrant methylation patterns are an early change in glioma tumorigenesis, earlier than genetic changes. Methylation within 5' regulatory CpG islands by DNA methyltransferase is one of the most common epigenetic modifications. 5-Aza-2' deoxycytidine (azacytidine) inhibits DNA methyltransferase and has been used in vitro to induce the expression of genes silenced by methylation. We have utilized azacytidine treatment and a micro-array system to investigate methylation influenced gene expression across several tumor cell lines of different lineage (brain, breast, prostate, liver). Using this system we have demonstrated that the expression of HAI-2/PB is under methylation control to a variable extent in glioma cell lines, in comparison to the other tested cell lines. Because the expression of HAI-2/B is inversely related to glioma invasiveness and degree of malignancy, this finding may provide insight into glioma initiation and progression as well as potentially providing new therapeutic targets. PMID- 14558598 TI - Prediction of pathology and survival by FDG PET in gliomas. AB - OBJECTIVES: Despite being in use for nearly two decades, the utility of [18F]2 fluoro-2deoxy-D-glucose positron emission tomography (FDG PET) in the evaluation and treatment of brain tumors remains controversial. We retrospectively analyzed all patients with histologically proven gliomas, between the years 1990 and 2000, who underwent FDG PET studies at various stages of their treatment and who were followed till either death or for a minimum period of 1 year in an attempt to bring resolution to this controversy. METHODS: All PET scans prior to 1997 were acquired on an ECAT 951/31 scanner in 2D. Scans since 1997 were obtained on a Siemens HR+ scanner in 3D mode. The majority of FDG PET scans were co-registered with the magnetic resonance imaging (MRI) scans to aid in diagnosis and therapy. Based on independent visual inspection, two board certified nuclear medicine physicians graded the highest activity level of the tumor using the metabolic grading: 0 = no uptake; 1 = uptake less or equal to normal white matter; 2 = uptake greater than normal white matter and less than gray matter; 3 = uptake equal to or greater than gray mater. The measure of association of lambda (lambda) was used to measure the strength of predictive ability of FDG PET for pathological grading of the gliomas. The Cox proportional hazards regression model was used to assess the significance of grade of uptake on survival. RESULTS: A total of 331 patients were analyzed of which 137 had a PET scan prior to histological diagnosis and therapeutic intervention (mean age = 46.5 years; M:F = 1.7:1). Eighty six percent (143/166) of the patients with low uptake (metabolic scores 0,1) had low-grade gliomas (grade I,II) and 14% (23/166) high grade gliomas (grade III,IV) on histologic examination. Ninety four percent (154/165) of the patients with high uptake (metabolic scores 2,3) on PET had high grade gliomas and 7% (11/165) had low-grade gliomas on histologic examination. The grade of uptake had increasing significance on survival as the level increased from 'low' to 'high' (P = 0.0009). Ninety four percent (156/166) of the patients with low uptake survived for > 1 year (median survival of 28 months) and 19% survived for > 5 years. Only 29% (48/165) of patients with high uptake survived for > 1 year, (median survival of 11 months) and none survived for > 5 years. Irrespective of when the scan showed a high uptake of FDG, before or after intervention, the prognosis following that scan was poor. CONCLUSIONS: Our observations confirm the utility of FDG PET as a prognostic tool for the histological grading and survival in patients with gliomas and appears to more than complement pathological grading. PMID- 14558599 TI - Feasibility of long-term intraventricular therapy with mafosfamide (n = 26) and etoposide (n = 11): experience in 26 children with disseminated malignant brain tumors. AB - Treatment options for leptomeningeal disseminated brain tumors are limited by the lack of effective drugs for intrathecal therapy of non-hematologic malignancies. We report on our experience with an intraventricular therapy consisting of mafosfamide, a preactivated cyclophosphamide derivative, and etoposide. Between May 1994 and 2002, 26 patients aged 2-19 years with various intensely pretreated disseminated brain tumors received intraventricular mafosfamide via an indwelling subcutaneous reservoir. Twenty-three of them received a dose of 20 mg. Mafosfamide was administered once or twice weekly until remission was achieved and every 2-6 weeks thereafter as maintenance therapy for a total of 736 administrations (2-63/patient). Since March 1998, two patients were switched to receive intraventricular etoposide and nine received etoposide alternating with mafosfamide. Etoposide was given at a dose of 0.5 mg x 5 d every 3-6 weeks for a total of 122 courses (1-29/patient). Immediate toxicities such as transient headaches, nausea, and vomiting occurred with mafosfamide but were manageable with premedication. Etoposide did not cause any discomfort. No long-term toxicities attributable to intrathecal therapy as evidenced by magnetic resonance imaging or neurologic evaluation were observed. Since all patients received some sort of concurrent anti-cancer therapy, the efficacy of intrathecal therapy cannot be assessed independently. However, seven of 13 patients evaluable for response by cerebrospinal fluid (CSF) cytology developed CSF dissemination under systemic chemotherapy and cleared their CSF only after administration of intrathecal mafosfamide. In conclusion, intraventricularly administered mafosfamide at a dose of 20 mg and etoposide at a dose of 0.5 mg x 5 d for patients over 2 years of age are feasible and safe and may produce responses. PMID- 14558601 TI - Late development of frontal prolactinoma after resection of pituitary tumor. AB - The unusual case of a patient with a frontal lobe prolactinoma developed 14 years after resection of a histologically benign pituitary adenoma is presented. Sixteen years after resection of this frontal mass, and 30 years after the first intervention, the patient is symptom-free and without evidence of new intracranial tumors. The possibility that frontal tumor resulted as the late proliferation of microscopic tumor seeding that took place in the course of the first surgery, is suggested. Although this case can be considered as a pituitary carcinoma, the clinical course supports that tumor seeding from pituitary tumors not necessarily indicates a poor prognosis. PMID- 14558600 TI - Brain metastases in patients with cancer of unknown primary. AB - Between January 1985 and December 2000, 916 patients with brain metastases were treated with whole brain radiation therapy (WBRT) at the Department of Radiotherapy, University Hospital Freiburg. In 47 patients, a primary tumor could not be identified (cancer of unknown primary (CUP)). Sixteen patients had a solitary brain metastasis, 31 patients presented with multiple brain metastases. Surgical resection was performed in 15 patients, biopsy alone in 12 patients. WBRT was applied with daily fractions of 2 or 3 Gy to a total dose of 50 or 30 Gy, respectively. According to the recursive partitioning analysis (RPA) classes of the Radiation Therapy Oncology Group for patients with brain metastases none of the patients met the criteria for Class I, 23 for Class II, and 24 for Class III. The median overall survival (OS) for all patients with brain metastases (n = 916) was 3.4 and 4.8 months for patients with CUP (p = 0.45). In patients with CUP (n = 47) the median OS for patients with a single brain metastasis was 7.3 versus 3.9 months for patients with multiple brain metastases (p = 0.05). Median OS for patients with a Karnofsky performance status (KPS) > or = 70 was 6.3 months versus 3.2 months for KPS < 70 (p = 0.01). At multivariate analysis performance status and resection status could be identified as independent prognostic factors for the OS. PMID- 14558602 TI - Meningeal melanocytoma: an aggressive course for a benign tumor. AB - A 79-year-old female presented with difficulty ambulating and was found to have weakness and hyperreflexia in the lower extremities. Magnetic resonance imaging (MRI) revealed a large T8-T9 intraspinal tumor. She underwent a thoracic laminectomy, and excision of an intradural extramedullary lesion. The surgical specimen was soft, black tissue that consisted of a moderately cellular, deeply pigmented tumor. The neoplastic cells proved to be melanocytic, and were devoid of overt features of anaplasia, i.e., prominent nuclear pleomorphism, necrosis, significant mitotic activity, and high proliferation indices. Four months postoperatively, MRI demonstrated focal areas of enhancement in the conus medullaris and in the fourth ventricle, indicating leptomeningeal spread. Subsequently, the patient underwent whole brain radiation. On repeat imaging, there was nodular enhancement of the fourth ventricle and throughout the spinal cord. Despite chemotherapy and radiation therapy, the disease advanced and the patient expired. Meningeal melanocytoma is a rare, histologically benign tumor with good prognosis. However, local aggressive behavior has been recorded, especially in cases of subtotal gross resection. On a literature review, there was one case of cranial posterior fossa meningeal melanocytoma with associated lesions in both suprarenal glands and the left kidney, but there were no cases with distant metastasis. In this report, we present an unusual case of spinal meningeal melanocytoma with diffuse spread throughout the craniospinal axis that proved to be fatal. PMID- 14558603 TI - Harvesting blood stem cells from cranial bone at craniotomy--a preliminary study. AB - Primary brain tumors seldom infiltrate into the cranium, even if they are invasive in the central nervous system. In this study, we examined whether blood stem cells can be harvested from cranial bone at craniotomy. Bone marrow cells in cranial bone were counted in 181 craniotomy specimens after staining with hematoxylin and eosin. Marrow volume was measured in 37 specimens using three dimensional computed tomography (CT). In 10 cases, viable cells collected from very small bone pieces at craniotomy were cultured to examine granulocyte/macrophage colony-forming units (CFU-GM). In 2 cases, bone marrow cells were practically harvested from removed bone at surgery. The weight of bone flap at craniotomy was 35.0 +/- 18.0 g. Bone flap marrow contained 1.5 x 10(9) cells/ml. CT examination showed that bone flap volume was 35.0 +/- 9.0 ml and marrow ratio was 65.1 +/- 13.5%. Thus, at craniotomy, a typical bone flap contained about 3.4 x 10(10) cells. Bone marrow cell count gradually decreased as subject age increased. The bone pieces obtained at craniotomy contained 3.1 +/- 3.4 x 10(6) cells/g, and CFU-GM count was 0.4 x 10(5) cells/g. In one case, we collected 3.4 x 10(8) cells, including 1.8 x 10(6) CFU-GM colonies. In another case, we collected 9.7 x 10(8) cells, including 4.8 x 10(6) CFU-GM colonies. These findings indicate that, at craniotomy, the number of blood stem cells in a typical bone flap is sufficient for autologous blood stem cell rescue. PMID- 14558604 TI - Loss of heterozygosity for loci on chromosome arms 1p and 10q in oligodendroglial tumors: relationship to outcome and chemosensitivity. AB - Oligodendroglial tumors frequently have deletions ofchromosomal loci on 1p and 19q. Loss of heterozygosity (LOH) of chromosome 10 may be a negative prognostic factor. We reviewed 23 patients with oligodendroglial tumors, to evaluate the frequency of 1p and 10q LOH and correlate with clinical outcome. Three loci (D1S402, D1S1172, MCT118) on 1p and 2 loci (D10S520 and D10S521) on 10q were analyzed for LOH using PCR techniques. Sixteen oligodendrogliomas (6 low grade and 10 anaplastic) and 7 oligoastrocytomas (1 low grade and 6 anaplastic) were studied. Overall 14/22 (64%) showed 1p LOH and 7/23 (30%) showed 10q LOH. Of 7 patients with some response to chemotherapy, all showed 1p LOH and none had 10q LOH. Of 5 patients with stable or progressive disease, 1 had 1p LOH and 2 showed 10q LOH. The presence of 1p LOH was significantly associated with response to chemotherapy (p = 0.02). Median progression free survival (PFS) was 31 months for 1p intact patients and 118 months for the 1p LOH group (p = 0.014). Median PFS for 10q LOH patients was 31 and 118 months for patients with intact chromosome 10 (p = 0.016).1p LOH is a predictor of response to chemotherapy and a good prognostic factor. 10q LOH is less common in oligodendroglial tumors but predicts for worse outcome. Molecular genotyping of oligodendroglial tumors is recommended as part of the standard diagnostic workup. PMID- 14558605 TI - Intra-cerebral schwannoma simulating glioma. AB - An intra-cerebral schwannoma, presenting as a cystic, calcified, enhancing frontal mass, arising in a 52-year-old woman was misdiagnosed as a glioma and treated with radiotherapy. This observation emphasizes the importance of careful histological reexamination of all brain tumors when a discrepancy appears between the initial histological diagnosis and the clinical evolution, in order to recognize rare curable entities and to avoid potentially toxic treatment. PMID- 14558606 TI - Enhancement of DNA micro-array analysis using a shear-driven micro-channel flow system. AB - A very simple micro-channel flow system is used to investigate the potential gain in hybridization rate stemming from the induction of a convective flow past the surface of a DNA micro-array. Reporting on a series of preliminary experiments wherein a two-dimensional convective flow is created past the surface of a conventional micro-array slide, the analysis time could be brought down from overnight waiting (16 h) to some 10 to 30 min. The experiments open the road towards the development of novel, convection-driven hybridization systems yielding shorter analysis times, and/or lower detection limits. PMID- 14558607 TI - Normalizing the background and removing the trend in one-dimensional DNA fingerprint images. AB - Maximizing an individual's genetic information from its DNA fingerprint image depends on the number of bands distinguished from the background. To approach this goal, the background should be normalized while the information is preserved. Morphological operators have been used by some authors to normalize the background for two-dimensional gel images. Methods such as mean, median and "maxpolygon" are presented in this work to normalize the background in DNA fingerprint images. Mean and median methods will lead to some deformations. Visual evaluation of the results show that the original shape of the column signals are better preserved by the maxpolygon. PMID- 14558608 TI - Analysis of site-directed mutagenesis constructs by capillary electrophoresis using linear polymer sieving matrices. AB - Site-directed mutagenesis is a novel molecular biology tool, which introduces mutations into DNA fragments of interest in a well-defined manner. Sequences with designed mutations can be generated in this way to express altered protein sequences for structure-function relationship studies. However, prior to gene expression, it is important to analyze the DNA construct to see whether the introduction of the mutation was indeed successful. Currently DNA sequencing is the method of choice for this verification. This paper introduces the combination of primer extension and capillary electrophoresis using linear polymer sieving matrices as an efficient alternative for this type of mutation analysis. The site directed mutagenesis construct served as template in the primer extension reaction that employed a fluorophore labeled primer in close proximity to the mutation. Appropriate ddNTP was used to block the extension when the mutation was present, while the other three dNTPs enabled elongation of the primer. Alternatively, non-labeled primers can be used with the proper fluorophore labeled ddNTPs to block the reaction. Rapid analysis of the labeled primer extension products (mutant or wild type) was obtained by capillary electrophoresis using denaturing sieving matrix and laser-induced fluorescence detection. PMID- 14558609 TI - Differential gene expression analysis by micro-preparative capillary gel electrophoresis. AB - Differential display analysis by cDNA fractionation, collection of differentially expressed fractions of interests and their downstream characterization is demonstrated. cDNA pools from two strains of Cochliobolus heterostrophus fungus were generated by specific restriction digestion and selective ligation. Micropreparative separation and isolation of differentially expressed transcript representatives were accomplished by high-performance capillary gel electrophoresis. The collected individual DNA molecules were polymerase chain reaction amplified and sequenced to create expressed sequence tags for the genes of interests. High resolving power and sensitivity of capillary gel electrophoresis enabled fast and automated processing of minute amounts of cDNA samples with high precision. PMID- 14558610 TI - Single nucleotide polymorphism analysis by allele-specific primer extension with real-time bioluminescence detection in a microfluidic device. AB - A microfluidic approach for rapid bioluminescent real-time detection of single nucleotide polymorphism (SNP) is presented. The method is based on single-step primer extension using pyrosequencing chemistry to monitor nucleotide incorporations in real-time. The method takes advantage of the fact that the reaction kinetics differ between matched and mismatched primer-template configurations. We show here that monitoring the initial reaction in real time accurately scores SNPs by comparing the initial reaction kinetics between matched and mismatched configurations. Thus, no additional treatment is required to improve the sequence specificity of the extension, which has been the case for many allele-specific extension assays. The microfluidic approach was evaluated using four SNPs. Three of the SNPs included primer-template configurations that have been previously reported to be difficult to resolve by allele-specific primer extension. All SNPs investigated were successfully scored. Using the microfluidic device, the volume for the bioluminescent assay was reduced dramatically, thus offering a cost-effective and fast SNP analysis method. PMID- 14558611 TI - Analysis of double-stranded DNA by microchip capillary electrophoresis using polymer solutions containing gold nanoparticles. AB - The impact of gold nanoparticles (GNPs) on the microchip electrophoretic separation of double-stranded (ds) DNA using poly(ethylene oxide) (PEO) is described. Coating of the 75-microm separation channel on a poly(methyl methacrylate) (PMMA) plate in sequence with poly(vinyl pyrrolidone), PEO, and 13 nm GNPs is effective to improve reproducibility and resolution. In this study, we have also found that adding 13-nm GNPs to 1.5% PEO is extremely important to achieve high resolution and reproducibility for DNA separation. In terms of the stability of the GNPs, 100 mM glycine-citrate buffer at pH 9.2 is a good buffer system for preparing 1.5% PEO. The separation of DNA markers V and VI ranging in size from 8 to 2176 base pairs has been demonstrated using the three-layer-coated PMMA microdevice filled with 1.5% PEO containing the GNPs. Using these conditions, the analysis of the polymerase chain reaction products of UGT1A7 was complete in 7 min, with the relative standard deviation values of the peak heights and migration times less than 2.3% and 2.0%, respectively. In conjunction with stepwise changes of the concentrations of ethidium bromide (0.5 and 5 microg/ml), this method allows improved resolution and sensitivity for DNA markers V and VI. PMID- 14558612 TI - Determination of itraconazole and hydroxyitraconazole in human serum and plasma by micellar electrokinetic chromatography. AB - The electrokinetic separation of the hydrophobic antimycotic drug itraconazole (ITC) and its major metabolite, hydroxyitraconazole (HITC), by a binary aqueous organic solvent medium containing sodium dodecylsulfate, by microemulsion electrokinetic chromatography (MEEKC) and by micellar electrokinetic chromatography (MEKC) was studied. The results suggest that the first approach is difficult to apply and that there is no substantial difference between separations performed using MEEKC and MEKC modified with n-butanol. The simpler MEKC method is more than adequate and was thus employed for the analysis of ITC and HITC in human serum and plasma. Separation was achieved in plain fused-silica capillaries having a low-pH buffer (pH 2.2) with sodium dodecyl sulfate micelles and reversed polarity. The addition of 2-propanol and n-butanol enhanced analyte solubility and altered the selectivity of the separation by influencing the magnitude of the electrophoretic component in the separation mechanism. Under optimised conditions and using head-column field-amplified sample stacking, an internal standard, ITC and two forms of HITC could be separated in under 9 min, with detection limits less than 0.01 microg/mL. Analysis of samples from patients currently prescribed ITC revealed a different HITC peak area ratio to that of the standards, suggesting a stereoselective component of ITC metabolisation. Comparison of MEKC data with those of a HPLC method employed on a routine basis showed excellent agreement, indicating the potential of this approach for therapeutic drug monitoring of ITC. PMID- 14558613 TI - Separation and identification of zaleplon metabolites in human urine using capillary electrophoresis with laser-induced fluorescence detection and liquid chromatography-mass spectrometry. AB - A capillary electrophoresis (CE) method using laser-induced fluorescence (LIF) detection for the determination of the hypnotic drug zaleplon and its metabolites in human urine could be developed using carboxymethyl-beta-cyclodextrin as a charged carrier. By the help of a complementary HPLC method coupled to mass spectrometry, three metabolites present in human urine could be identified as 5 oxozaleplon, 5-oxo-N-deethylzaleplon and 5-oxozaleplon glucuronide. N Deethylzaleplon, a previously described zaleplon metabolite, as well as zaleplon itself could not be detected in human urine by the CE-LIF assay. The results were confirmed by spiking with reference compounds of the phase I metabolites. The metabolites differed very much concerning their fluorescence intensities, thus the 5-oxo metabolites present as lactam tautomer fluoresced tenfold lower than the unchanged drug zaleplon and its N-deethylated metabolite. The glucuronide of the 5-oxozaleplon, however, showed high fluorescence due to its lactim structure. Limits of quantification yielded by the CE-LIF assay including a ten-fold preconcentration step by solid-phase extraction were 10 ng/ml for zaleplon and N deethylzaleplon and 100 ng/ml for 5-oxozaleplon and 5-oxo-N-deethylzaleplon. PMID- 14558614 TI - Analysis of Chinese medicine preparations by capillary electrophoresis-mass spectrometry. AB - In Chinese medicines, herbs are usually prepared before use by patients. Since the preparation procedures convert the original component into one or more products, study of the procedures is usually complex and involves several compounds. On-line coupling of capillary electrophoresis (CE) to mass spectrometry (MS) allows both the efficient separation of CE and the specific and sensitive detection of MS to be achieved. In this study, CE-MS was applied to the determination of alkaloids in Maqianzi (the seed of Strychnos pierrian) and Wutou (aconite root, Radix aconiti praeparata) during the preparation procedure. With optimal CE-MS conditions, alkaloids in both prepared and unprepared Maqianzi were determined successfully in the total ion current (TIC) mode. However, single ion monitoring (SIM) had to be applied for the separation of aconitum alkaloids and their hydrolysis products. Quantification data indicated that MS detection under SIM mode is more sensitive than UV detection. Based on the CE-MS method developed, the hydrolysis of aconitum alkaloids in water and methanol was also studied. PMID- 14558615 TI - Simultaneous determination of methotrexate and its eight metabolites in human whole blood by capillary zone electrophoresis. AB - A simple and selective capillary electrophoretic method was established for the simultaneous determination of methotrexate (MTX), 7-hydroxymethotrexate (7 OHMTX), 2,4-diamino-N10-methylpteroic acid (DAMPA), and polyglutamate derivatives [MTX-(Glu)n, n=2-7] in whole blood. After extraction, those analytes were separated by fused-silica capillary and a running buffer containing glycine (1.2 M, pH 9.3). The quantitative ranges were 10-50 microM for each analyte. The intra and inter-day RSD and RE values were all less than 6 and 11%, respectively. The limits of detection (S/N= 3, injection 5 s) were found to be 1 microM for MTX, 7 OHMTX, MTX-(Glu)2, MTX-(Glu)3, and MTX-(Glu)4; 3 microM for MTX-(Glu)5 and MTX (Glu)6; 5 microM for MTX-(Glu)7, and 8 microM for DAMPA. All recoveries were greater than 94%. This method was applied to blood MTX monitoring in a patient with acute lymphoblastic leukemia. PMID- 14558616 TI - Separation and on-line concentration of bisphenol A and alkylphenols by micellar electrokinetic chromatography with anionic surfactant. AB - Separation and on-line concentration of bisphenol A and three alkylphenols were investigated by micellar electrokinetic chromatography with the anionic surfactant, sodium dodecyl sulfate. The separation conditions were optimized by the simultaneous addition of the organic solvent and cyclodextrin to the running solution. The separation of hydrophobic analytes and 4-nonylphenol isomers was improved by the addition of 10% methanol and 5 mM beta-cyclodextrin to the running solution. When the sweeping with the running solution was used as the on line concentration procedure, 69-, 48-, 55- and 41-fold increases in detection sensitivity were obtained for bisphenol A, 4-tert.-butylphenol and 4-(1,1,3,3 tetramethylbutyl)phenol, and the second peak of 4-nonylphenol isomers, respectively. The detection limits were 0.0071, 0.0065, 0.021 and 0.055 mg/l, respectively. These results were better than those with the cationic surfactant, tetradecyltrimethylammonium bromide. PMID- 14558617 TI - Investigation of preconcentration strategies for the trace analysis of multi residue pesticides in real samples by capillary electrophoresis. AB - In this work, on-line preconcentration strategies were investigated for the multi residue analysis of pesticides in drinking water and vegetables using micellar electrokinetic chromatography. Among the on-line strategies, sweeping and stacking with reverse migration of micelles (SRMM), with and without the insertion of a plug of water before sample injection, were contrasted. A new version of SRMM was also introduced. The modification consisted of momentarily applying a positive voltage at the inlet vial right after sample has been injected, increasing the efficiency by which the analytes are captured. Nine pesticides from different classes, carbendazim (benzimidazole), simazine, atrazine, propazine and ametryn (triazine), diuron and linuron (urea), carbaryl and propoxur (carbamate), were baseline separated in less than 6 min with a electrolyte composed of 20 mmol l(-1) phosphate buffer at pH 2.5, containing 25 mmol l(-1) sodium dodecyl sulfate and 10% methanol. Limits of detection (LODs) in the order of 2-46 microg l(-1) for the pesticides under investigation were obtained solely using the on-line strategies. Enrichment factors of 3-18-fold were obtained. These factors were computed as the improvement of the concentration LODs with respect to the reference condition (injection of 10 s at 2.5 kPa pressure). The proposed methodologies were applied to the analysis of pesticides in complex matrices such as carrot extracts where the detection of 2.5 microg l(-1) was illustrated. By combining off-line solid-phase extraction and the proposed on-line strategies, the detection of pesticides in drinking water at the 0.1 microg l(-1) level was conceived. PMID- 14558618 TI - Separation and characterization of humic acids from Antarctica by capillary electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Inclusion complexes of humic acids with cyclodextrins. AB - A new capillary electrophoresis procedure based on micellar electrokinetic capillary chromatography for the separation of humic acids (HAs) isolated from Antarctica soil was developed. The HAs were separated and characterized using a background electrolyte containing 0.09 M borate+0.09 M Tris+0.001 M EDTA (BTE) of pH 8.3, modified with alpha-, beta-, or gamma-cyclodextrins (CDs) and sodium dodecyl sulfate. It was found that from alkaline solution of HAs in the presence of CDs, the HAs are not completely precipitated with a strong acid and a certain part (some fractions) remains soluble. Mass spectrometry shows that HAs contain 15-25 simple low-Mr compounds and several families of compounds with similar structure (m/z approximately 800-1200). Comparison of HA analysis from Antarctica soil with those of soil HAs from the American continent show a high similarity between the samples and confirm several identical compounds and some with very similar structural units. PMID- 14558619 TI - Potential of microemulsion electrokinetic chromatography for the separation of priority endocrine disrupting compounds. AB - This work examines the potential of microemulsion electrokinetic chromatography for the separation of several priority endocrine disrupting compounds (EDCs). The optimised microemulsion system comprised 25 mM phosphate buffer pH 2, 80 mM octane, 900 mM butanol, 200 mM sodium dodecyl sulphate and was further modified with 20% propanol. The use of a low pH buffer resulted in the suppression of electroosmotic flow within the capillary. Reversal of the conventional electrode polarity resulted in faster migration of hydrophobic compounds. Test analytes included the octylphenol, nonylphenol and nonylphenol diethoxylate, which are breakdown products of the alkylphenolic detergents. The synthetic oestrogens diethylstilbestrol and ethynyloestradiol were also included in the separation along with the plastic monomer bisphenol-A. Test analytes were selected due to their reported presence in environmental samples namely industrial and domestic wastewater treatment effluents and sludges. Using the optimised method a separation of six EDCs was achieved within 15 min. The optimised method was then applied to the analysis of a spiked wastewater influent sample with UV detection of all six compounds at 214 nm. PMID- 14558620 TI - Novel modes of capillary electrophoresis for the determination of endocrine disrupting chemicals. AB - The synthesis and usage of a wide range of organic chemicals has increased dramatically over the last five decades. These compounds sometimes termed endocrine disrupting chemicals include agricultural pesticides, industrial solvents, dyes, plasticisers, detergents and heat exchangers. Concerns have been raised about the potential adverse effects of these compounds on humans and wildlife species. Our objectives are to develop a method to identify, using novel capillary electrophoretic techniques, the endocrine disrupting compounds that are reported to be present in environmental samples. The CE modes, capillary zone electrophoresis, micellar electrokinetic chromatography (MEKC), cyclodextrin modified MEKC (CD-MEKC) and electroosmotic flow-suppressed CD-MEKC were investigated for the determination of a range of endocrine disrupting chemical compounds. This paper shows some initial results obtained. PMID- 14558621 TI - Stability studies of propoxur herbicide in environmental water samples by liquid chromatography-atmospheric pressure chemical ionization ion-trap mass spectrometry. AB - Liquid chromatography-atmospheric pressure ionization ion-trap mass spectrometry has been investigated for the analysis of polar pesticides in water. The degradation behavior of propoxur, selected as a model pesticide belonging to the N-methylcarbamate group, in various aqueous matrices (Milli-Q water, drinking water, rain water, seawater and river water) was investigated. Two interfaces of atmospheric pressure ionization, atmospheric pressure chemical ionization (APCI) and electrospray ionization (ESI), were compared during the study. Propoxur and its transformation product (N-methylformamide) were best ionized as positive ions with both APCI and ESI, while another transformation product (2-isopropoxyphenol) yielded stronger signals as negative ions only with APCI. In addition, the effects of various pH, matrix type and irradiation sources (sunlight, darkness, indoor lighting and artificial UV lamp) on the chemical degradation (hydrolysis) were also assessed. From the kinetic studies of degradation, it was found that the half-life of propoxur was reduced from 327 to 161 h in Milli-Q water with variation of irradiation conditions from dark to sunlight exposure. Degradation rates largely increased with increasing pH. The half-life of the target compound dissolved in Milli-Q water under darkness decreased from 407 to 3 h when the pH of Milli-Q water was increased from 5 to 8.5. These suggest that hydrolysis of propoxur is light-intensity and pH-dependent. In order to mimic contaminated natural environmental waters, propoxur was spiked into real water samples at 30 microg/l. The degradation of propoxur in such water samples under various conditions were studied in detail and compared. With the ion trap run in a time scheduled single ion monitoring mode, typical limits of detection of the instrument were in the range of 1-10 microg/l. PMID- 14558622 TI - Optimization of microwave-assisted extraction and supercritical fluid extraction of carbamate pesticides in soil by experimental design methodology. AB - Orthogonal array design (OAD) was applied for the first time to optimize microwave-assisted extraction (MAE) and supercritical fluid extraction (SFE) conditions for the analysis of four carbamates (propoxur, propham, methiocarb, chlorpropham) from soil. The theory and methodology of a new OA16 (4(4)) matrix derived from a OA16 (2(15)) matrix were developed during the MAE optimization. An analysis of variance technique was employed as the data analysis strategy in this study. Determinations of analytes were completed using high-performance liquid chromatography (HPLC) with UV detection. Four carbamates were successfully extracted from soil with recoveries ranging from 85 to 105% with good reproducibility (approximately 4.9% RSD) under the optimum MAE conditions: 30 ml methanol, 80 degrees C extraction temperature, and 6-min microwave heating. An OA8 (2(7)) matrix was employed for the SFE optimization. The average recoveries and RSD of the analytes from spiked soil by SFE were 92 and 5.5%, respectively except for propham (66.3+/-7.9%), under the following conditions: heating for 30 min at 60 degrees C under supercritical CO2 at 300 kg/cm2 modified with 10% (v/v) methanol. The composition of the supercritical fluid was demonstrated to be a crucial factor in the extraction. The addition of a small volume (10%) of methanol to CO2 greatly enhanced the recoveries of carbamates. A comparison of MAE with SFE was also conducted. The results indicated that >85% average recoveries were obtained by both optimized extraction techniques, and slightly higher recoveries of three carbamates (propoxur, propham and methiocarb) were achieved using MAE. SFE showed slightly higher recovery for chlorpropham (93 vs. 87% for MAE). The effects of time-aged soil on the extraction of analytes were examined and the results obtained by both methods were also compared. PMID- 14558623 TI - Separation of phenols as neutral compounds by micellar electrokinetic capillary chromatography. AB - Separation of phenols as neutral solutes by micellar electrokinetic capillary chromatography provides a quantitative linear dynamic range of 6000-13,000. Since the compounds are injected and separated as neutral solutes, the dispersive processes of anti-stacking and electrodispersion are eliminated. Optimized conditions allow for sub-ppm quantitation of trace impurities in the presence of the major components at various stages of the production of high purity phenols. The background electrolyte consists of 100 mM sodium dodecyl sulfate in 50 mM phosphate buffer pH 7. The method is precise, reliable, and the limits of detection are superior compared to HPLC by a factor of 20. PMID- 14558624 TI - New method for analyzing the nitrite level in PC12 cells using capillary electrophoresis. AB - Nitric oxide (NO) is a gaseous molecule shown to have signal transmitter properties in organisms. Direct measurement of NO in physiological conditions has been difficult due to its short lifetime and low concentration. Nitrite has been used as a marker for NO formation in biological systems. Capillary electrophoresis (CE) has been recently used to measure nitrite in biological fluids. The purpose of this study is to analyze nitrite in PC12 cells (pheochromocytoma cell line) using CE. Optimal CE performance was employed with 150 mM Tris-phosphate, 6 microM hexadecyltrimethyammonium chloride buffer at pH 7.0 and a fused-silica column of 57 cm x 75 microm I.D. The signal was measured with a UV detector at 214-nm wavelength and negative potential of 10 kV was applied for nitrite analysis. Under the optimum conditions, we monitored the changes in the concentration of the nitrite levels through synergistic stimulation of tumor necrosis factor alpha plus gamma-interferon in PC12 cells. PMID- 14558625 TI - Development of a novel running buffer for the simultaneous determination of nitrate and nitrite in human serum by capillary zone electrophoresis. AB - In order to improve NO2- peak height and obtain a convenient buffer system for the assay of nitrogen monooxide metabolites, we developed a novel running buffer for the simultaneous determination of nitrite and nitrate in human serum by capillary electrophoresis. The addition of cetyltrimethylammonium chloride to the running buffer resulted in high-speed separation using reverse electroosmotic flow. Highly sensitive determination was also achieved using stacking with 10 fold diluted sample solutions. The samples were injected hydrodynamically for 100 s into a 50 cm x 75 microm I.D. capillary. The separation voltage was 10 kV (negative polarity). UV detection was performed at 214 nm. We obtained complete separation of nitrite and nitrate in deproteinized human serum within 6 min with optimum analytical conditions. Linear calibration curves for nitrite and nitrate for both peak height and peak area were obtained with standard addition method. The limits of detection obtained at a signal-to-noise ratio of 3 for nitrite and nitrate were 4.1 and 2.0 microM, while the values of relative standard deviation of peak height were 2.4 and 2.6%, respectively. PMID- 14558626 TI - Speciation of chromium by in-capillary reaction and capillary electrophoresis with chemiluminescence detection. AB - A sensitive method for the simultaneous determination of chromium(III) (Cr3+) and chromium(VI) (CrO4(2-)) using in-capillary reaction, capillary electrophoresis (CE) separation and chemiluminescence (CL) detection was developed. The chemiluminescence reaction was based on luminol oxidation by hydrogen peroxide in basic aqueous solution catalyzed by Cr3+ ion followed by capillary electrophoresis separation. Based on in-capillary reduction, chromium(VI) can be reduced by acidic sodium hydrogensulfite to form chromium(III) while the sample is running through the capillary. Before the electrophoresis procedure, the sample (Cr3+ and CrO4(2-)), buffer and acidic sodium hydrogensulfite solution segments were injected in that order into the capillary, followed by application of an appropriate running voltage between both ends. As both chromium species have opposite charges, Cr3+ ions migrate to the cathode, while CrO4(2-) ions, moving in the opposite direction toward the anode, react with acidic sodium hydrogensulfite which results in the formation of Cr3+ ions. Because of the migration time difference of both Cr3+ ions, Cr(III) and Cr(VI) could be separated. The running buffer was composed of 0.02 mol l(-1) acetate buffer (pH 4.7) with 1 x 10(-3) mol l(-1) EDTA. Parameters affecting CE-CL separation and detection, such as reductant (sodium hydrogensulfite) concentration, mixing mode of the analytes with CL reagent, CL reaction reagent pH and concentration, were optimized. The limits of detection (LODs) of Cr(III) and Cr(VI) were 6 x 10(-13) and 8 x 10(-12) mol l(-1) (S/N=3), respectively. The mass LODs for Cr(III) and Cr(VI) were 1.2 x 10(-20) mol (12 zmol) and 3.8 x 10(-19) mol (380 zmol), respectively. PMID- 14558628 TI - Capillary electrophoretic separation of heparin oligosaccharides under conditions amenable to mass spectrometric detection. AB - A capillary electrophoresis method for the separation of high-molecular-mass heparin oligosaccharides compatible with mass spectral detection was developed. Structurally defined heparin oligosaccharides ranging in size from tetrasaccharide to tetradecasaccharide were used to optimize the conditions. Applying normal and reversed polarity modes, these oligosaccharides were separated by CE under various conditions. Ammonium hydrogencarbonate (30 mM at pH 8.50) used as the running electrolyte system gave good separation efficiency and resolution in the normal polarity mode. Application of this method to the separation of complicated heparin oligosaccharide mixtures required the addition of electrolyte additives. Ammonium hydrogencarbonate (30 mM), containing triethylamine (10 mM), was useful for the separation of complex oligosaccharide mixtures. Run-to-run and day-to-day precision and limits of detection were established for these separations. PMID- 14558629 TI - Honoring donors and donor families. PMID- 14558627 TI - Separation of a complex mixture of heparin-derived oligosaccharides using reversed-phase high-performance liquid chromatography. AB - A reversed-phase ion-pairing high-performance liquid chromatography (RP-HPIPC) method for the separation of a complex mixture of heparin-derived oligosacchrides has been developed by a stepwise optimization of the mobile phase, in which the concentration of ion-pairing reagent, mobile phase pH, and acetonitrile concentration were varied. The resolution of more than 30 oligosaccharide components was obtained, under optimized conditions, in an analysis time of less than 30 min. This represents the first RP-HPLC method that can separate a complex mixture of both small and large sulfated oligosaccharides in a single chromatographic step. The heparin-derived oligosaccharides, in this mixture, can also be separated under a second set of RP-HPIPC conditions using a volatile ion pairing reagent, tributylammonium acetate, to aid in the recovery of individual sulfated oligosaccharides. Moreover, it was possible to replace sodium chloride gradient, required for eluting highly sulfated oligosaccharides, with a fixed, low concentration of a volatile salt, ammonium acetate, by utilizing an acetonitrile gradient. This solvent system might make it possible to directly interface this RP-HPIPC separation with mass spectral analysis. PMID- 14558630 TI - Religious altruism and the living organ donor. AB - Potential living organ donors who describe religious motivations as the basis for their donation, particularly in the absence of an emotional or genetic connection to the intended recipient, raise challenges for the transplant team who are more comfortable in assessing relational motivations than religious ones. This article provides a composite case scenario to illustrate some of the biases that may interfere with the assessment of the donor motivated by religious altruism and how religious altruism may be pathologized. The clinical challenges in assessing the religiously altruistic potential living donor are reviewed. Religious approaches to altruism and their implications for living organ donation are reviewed for a sample of the major world religions. PMID- 14558631 TI - Home monitoring for lung transplant candidates. AB - Home monitoring by lung transplant recipients has been effective for early detection of clinical problems. This study used an electronic diary for home monitoring by lung transplant candidates to improve communication between candidates and the transplant team. Candidates were randomized into control (52 subjects following standard telephone reporting procedures) and intervention (67 subjects using an electronic diary to record and transmit a range of health related measures) groups. Outcome measures were monitoring adherence and level of communication (for monitor acceptability and utilization), hospital length of stay after transplantation and survival at 4 months (for clinical effectiveness). Subjects used the diary without difficulty and with good adherence. Subjects and coordinator contacts were similar between groups; intervention group subjects were positive regarding contact based on diary use. There were no significant differences in clinical outcomes between groups. Changing diary questions might improve the effectiveness of electronic monitoring for lung transplant candidates. PMID- 14558632 TI - Avoidant coping with health problems is related to poorer quality of life among lung transplant candidates. AB - CONTEXT: Lung transplant candidates face numerous health-related stressors. Although previous work has described the range of coping strategies candidates may use, whether those strategies are related to quality of life in physical functioning, emotional, and social domains has rarely been examined. METHODS: Adult lung transplant candidates (N = 128) participated in semistructured interviews that included questions regarding global and domain-specific quality of life and a multidimensional assessment of coping with health-related problems. Demographic characteristics, health status, and other psychosocial variables were also assessed, and their effects were examined and controlled in multivariate analyses of the coping-quality-of-life relationship. RESULTS: Respondents were most likely to use active, acceptance, and support-seeking strategies to cope with health problems. Self-blame or avoidance were rarely used. Although used least often, avoidant coping was the most strongly and consistently related to quality of life. With demographic and psychosocial variables controlled, higher avoidant coping remained associated with significantly poorer global quality of life, bodily pain, difficulty with daily work or activities as a function of emotional problems, and depressive symptomatology. Avoidant coping accentuated the association of poor health status and lower quality of life. Among respondents with lower health status, using more avoidant coping was associated with greater difficulty functioning in daily activities as a result of emotional problems. Those with higher health status had less difficulty functioning in daily activities as a result of emotional problems. CONCLUSIONS: Transplant team members are encouraged to be aware of and help patients refrain from using avoidant coping strategies. PMID- 14558633 TI - Reproductive and contraceptive characteristics of premenopausal kidney transplant recipients. AB - OBJECTIVE: To obtain information on menstrual patterns before and after transplantation, desire for future pregnancy, and use of contraception among premenopausal kidney transplant recipients. STUDY DESIGN: This observational study collected information using self-administered anonymous questionnaires during a routine outpatient clinic visit. RESULTS: Of the 107 women who completed the questionnaire, 41 identified themselves as being premenopausal. Among the 41 premenopausal women, approximately half of the women reported their current menstrual patterns as normal and 26% were not using any form of contraception. Overall, 10 women (24%) reported a desire to become pregnant and 4 women (10%) had a successful pregnancy after transplantation. Most of the women who desired a future pregnancy (8/10) were receiving an immunosuppressive regimen that included mycophenolate mofetil. CONCLUSION: Kidney transplantation in the current era is associated with a return of normal menstrual function in the majority of female transplant recipients. A substantial fraction of women desire pregnancy after transplantation and many are using an immunosuppressive drug with limited safety data on use during pregnancy. More caution should be used with the use of newer immunosuppressive agents in sexually active premenopausal transplant recipients until more safety data are available. PMID- 14558634 TI - Clinical integration of billing for a pediatric nephrology and transplant program. AB - PURPOSE: To develop and implement a billing process that fully integrates all activities of a pediatric nephrology and transplant program, by facilitating and coordinating data from patients, physicians, hospitals, and third-party billing services to maximize revenues. METHODS: Financial operations were analyzed via a randomized audit of patient charts that focused on office procedures and revenue collection. Results based on monthly reports documenting revenue received and outstanding, procedures billed, and patient registration accuracy. RESULTS: The combination of improvements in patient registration, chart documentation, new billing sheets with procedure and diagnosis codes, physician in-service education, upgraded charges, and the recredentialing of all practice physicians realized an increase in revenue collections from 18% in 2000 to 89% in 2001. CONCLUSION: The need to integrate and coordinate information is vital for both billing accuracy and revenue collections. Integration of clinical services and billing procedures has maximized performance, profitability, and accuracy while decreasing administrative time and costs. PMID- 14558635 TI - Exploring donors' and recipients' attitudes about living donor kidney transplantation. AB - CONTEXT: There is a growing interest in living donor kidney transplantation because of its potential to reduce the current kidney shortage. OBJECTIVE: To explore the experience of potential recipients, recipients, potential donors, and donors with regard to living donor kidney transplantation and laparoscopic living donor nephrectomy. METHOD: Twenty-five patients participated in a series of focus group interviews. The interviews were tape-recorded and a content analysis was performed. RESULTS: All participants had a positive attitude toward living donor kidney transplantation and laparoscopic living donor nephrectomy, though this procedure was not a factor that influenced potential donors' decision to donate a kidney. None of the potential donors and donors felt coerced into giving a kidney; they were motivated by a desire to help a loved one who was suffering. The main perceived impediment to donate a kidney was the potential recipients' reluctance to accept the offer. Potential recipients and recipients found it difficult to ask for a kidney and worried about the consequences of a kidney removal on their donor's health. A potential barrier to living donor kidney transplantation was that recipients would never have accepted a kidney if their donor had to pay for the kidney removal. In addition, recipients believed that a kidney from a non-blood-related donor was not as a good a match as a kidney from a blood-related donor. CONCLUSION: This exploratory study reveals that donors, potential donors, recipients, and potential recipients had an overall positive attitude toward living donor kidney transplantation and laparoscopic living donor nephrectomy. This study also sheds light on a number of barriers that have not been previously reported. PMID- 14558636 TI - Culture, brain death, and transplantation. AB - From the social sciences, we know the space between life and death is historically and culturally constructed, fluid and open to dispute. The definition of death has cultural, legal, and political dimensions. As healthcare becomes more culturally diverse, the interface between culture and the delivery of healthcare will increase. In our increasingly pluralistic, interdependent society, there is a growing demand to integrate healthcare, including transplantation, into a broader context that respects both individual and cultural diversity. It is important that we first consider and explore what elements of Western healthcare practices including definitions and advances, such as brain death and organ donation, are culturally influenced. This article highlights some of the cultural influences on brain death by focusing on Western and Japanese perspectives on the permissibility of organ procurement from brain dead persons. It also offers 4 recommendations for healthcare workers working cross-culturally. PMID- 14558637 TI - Families' understanding of brain death. AB - CONTEXT: Families' understanding of brain death may be a factor that contributes to decisions regarding organ donation. OBJECTIVE: To examine factors related to families' understanding of brain death and how those factors affect families' decisions about organ donation. DESIGN: Chart reviews were conducted on all deceased patients at 9 trauma hospitals. Interviews were then conducted with healthcare providers who spoke with families about organ donation and family decision makers. SETTING: Southwest Pennsylvania and Northeast Ohio. PARTICIPANTS: The sample consisted of 403 families of organ donor-eligible patients who reported that someone at the hospital told them the patient was brain dead. MAIN OUTCOME MEASURES: Families' awareness of brain-death testing procedures, understanding of brain death, and donation decision. RESULTS: Most families (96%) were told their family member was brain dead, but only 28.3% were able to provide a completely correct definition of brain death. No association between the definition of brain death and willingness to donate was found. Those who donated were less likely to agree with the statement that someone is dead only when the heart stops than those who did not donate. Respondents who accepted that their loved one was dead when informed the patient was brain dead were more likely to donate than those who did not consider the patient dead until mechanical support was turned off and the heart stopped. PMID- 14558638 TI - Development of a successful non-heart-beating organ donation program. AB - To help alleviate the organ shortage, transplant centers are using organs from expanded-criteria donors, who were considered unsuitable just a few years ago, such as non-heart-beating donors. In 1998, we made a concerted effort to increase the number of non-heart-beating donors recovered by our organ procurement organization. In this paper, we discuss the steps in establishing this program, including transplant center support, estimating the number of potential non-heart beating donors, organ procurement support, protocol development, hospital development, education, putting the protocol into practice, follow-up, and effect of the program on organ procurement. With the establishment of this program, the number of non-heart-beating donors increased from 2% to 5% per year to over 10% for the past 2 years. From these donors, 61 of 82 recovered kidneys were transplanted into 58 patients, and 18 of 20 recovered livers were transplanted. A non-heart-beating donor program can significantly add to the number of organ transplants and successful transplantations. PMID- 14558639 TI - The epidemiology and natural history of hepatitis C virus infection. AB - Hepatitis C virus (HCV) infection is the most common chronic blood-borne infection in the United States. HCV infection is generally benign in its acute stage but tends to become chronic in more than 70% of patients, at which stage it can induce liver cirrhosis, end-stage liver disease, and hepatocellular carcinoma. Approximately 2.7 million Americans are estimated to have chronic HCV infection. Although the incidence of HCV infection is believed to be falling, the prevalence of HCV-related liver disease is rising. Better identification of risk factors for HCV transmission and improved understanding of the infection's natural history should refine measures for preventing the spread of infection and preventing complications in those infected. PMID- 14558640 TI - Tests and screening strategies for the diagnosis of hepatitis C. AB - The threshold for testing for hepatitis C virus (HCV) should be low for persons with any risk factor for HCV infection. Current practice calls for first screening for antibodies to HCV and then testing for HCV RNA in those in whom antibodies are detected. Viral testing can distinguish between active and resolved HCV infection and also determine viral load, which can help predict response to antiviral therapy. Many highly sensitive assays are available for testing for HCV RNA. Once HCV infection is diagnosed, the HCV genotype should be determined to help predict treatment response and duration. Liver biopsy can aid in disease staging and help guide treatment decisions. Practical and efficient screening strategies for HCV are guided by risk factors for HCV infection. PMID- 14558641 TI - Management of newly diagnosed hepatitis C virus infection. AB - All patients with chronic hepatitis C virus infection are potential candidates for antiviral therapy. Careful patient selection can optimize the response to therapy and enhance safety. Pegylated forms of interferon, when combined with ribavirin, can "cure" the majority of patients undergoing therapy, and these agents have become the new standard of care for chronic hepatitis C. Careful and timely management of side effects, which are experienced by all patients, can improve adherence to antiviral therapy and further improve response rates. PMID- 14558642 TI - Treatment options for nonresponders and relapsers to initial therapy for hepatitis C. AB - As treatment for chronic hepatitis C virus (HCV) infection has advanced over the past decade, efforts have evolved to retreat patients who did not achieve a sustained virologic response to previous antiviral regimens. Retreating nonresponders to interferon alfa monotherapy with a combination of interferon and ribavirin yields a sustained virologic response in 9% to 32% of patients, whereas retreatment with peginterferon alfa plus ribavirin yields a sustained virologic response in up to 30% to 40% of patients. Sustained virologic response is more likely in retreated patients with HCV genotype 2 or 3, low serum HCV RNA levels, and lack of response to prior interferon monotherapy. Retreatment of nonresponders to interferon-ribavirin combination therapy is associated with lower response rates (< or = 20%). Despite treatment advances, the efficacy of current antiviral regimens for nonresponders remains inadequate. The next few years will see more-targeted antiviral regimens for these patients and therapies focused on slowing the progression of liver disease rather than viral eradication. PMID- 14558643 TI - Special management challenges in hepatitis C. AB - Infection with hepatitis C virus (HCV) often coexists with other conditions and patient factors that complicate its management. Infection with HIV is a particularly widespread and vexing comorbidity of HCV infection, since HIV facilitates HCV transmission and renders HCV more opportunistic. This review provides a practical overview of major comorbidities and patient factors that require special management considerations in patients with HCV infection. PMID- 14558645 TI - Analysis of contact mechanics in McKee-farrar metal-on-metal hip implants. AB - Contact mechanics analysis for a typical McKee-Farrar metal-on-metal hip implant was carried out in this study. The finite element method was used to predict the contact area and the contact pressure distribution at the bearing surfaces. The study investigated the effects of the cement and underlying bone, the geometrical parameters such as the radial clearance between the acetabular cup and the femoral head, and the acetabular cup thickness, as well as other geometrical features on the acetabular cup such as lip and studs. For all the cases considered, the predicted contact pressure distribution was found to be significantly different from that based upon the classical Hertz contact theory, with the maximum value being away from the centre of the contact region. The lip on the cup was found to have a negligible effect on the predicted contact pressure distribution. The presence of the studs on the outside of the cup caused a significant increase in the local contact pressure distribution, and a slight decrease in the contact region. Reasonably good agreement of the predicted contact pressure distribution was found between a three-dimensional anatomical model and a simple two-dimensional axisymmetric model. The interfacial boundary condition between the acetabular cup and the underlying cement, modelled as perfectly fixed or perfectly unbonded, had a negligible effect on the predicted contact parameters. For a given radial clearance of 0.079 mm, the decrease in the thickness of the acetabular cup from 4.5 to 1.5 mm resulted in an increase in the contact half angle from 15 degrees to 26 degrees, and a decrease in the maximum contact pressure from 55 to 20 MPa. For a given acetabular cup thickness of 1.5 mm, a decrease in the radial clearance from 0.158 to 0.0395 mm led to an increase in the contact half-angle from 20 degrees to 30 degrees, and a decrease in the maximum contact pressure from 30 to 10 MPa. For zero clearance, although the contact pressure was significantly reduced over most of the contact area, the whole acetabular cup came into contact with the femoral head, leading to stress concentration at the edge of the cup. Design optimization of the geometrical parameters, in terms of the acetabular cup thickness and the radial clearance, is important, not only to minimize the contact stress at the bearing surfaces, but also to avoid equatorial and edge contact. PMID- 14558644 TI - A different fixation of the femoral component in total knee arthroplasty may lead to preservation of femoral bone stock. AB - Good femoral bone stock is important for the stability of the femoral component in revision knee arthroplasty. However, the primary total knee replacement (TKR) may cause significant loss of bone stock in the distal anterior femur. Earlier stress-induced bone remodelling simulations have suggested that a completely debonded component may save bone stock in the distal anterior region. However, these simulations did not consider the fixation of a debonded implant and possible secondary effects of micromotions and osteolysis at the interface. The current study tries to combine the preservation of bone stock with adequate component fixation. Different bone remodelling simulations were performed around femoral knee components with different sizes of bonding area and different friction characteristics of the debonded area. The fixation of the femoral component with different bonding characteristics is quantified with calculated implant-bone interface stresses. The results show that a bonded femoral component with a debonded inner side of the anterior flange may significantly reduce bone resorption in the endangered distal anterior femur, without jeopardizing the fixation of the femoral implant. This effect may be obtained in vivo by using a femoral component with a highly polished inner side of the anterior flange. PMID- 14558646 TI - A finite element model of an idealized diarthrodial joint to investigate the effects of variation in the mechanical properties of the tissues. AB - The stiffness of articular cartilage increases dramatically with increasing rate of loading, and it has been hypothesized that increasing the stiffness of the subchondral bone may result in damaging stresses being generated in the articular cartilage. Despite the interdependence of these tissues in a joint, little is understood of the effect of such changes in one tissue on stresses generated in another. To investigate this, a parametric finite element model of an idealized joint was developed. The model incorporated layers representing articular cartilage, calcified cartilage, the subchondral bone plate and cancellous bone. Taguchi factorial design techniques, employing a two-level full-factorial and a four-level fractional factorial design, were used to vary the material properties and thicknesses of the layers over the wide range of values found in the literature. The effects on the maximum values of von Mises stress in each of the tissues are reported here. The stiffness of the cartilage was the main factor that determined the stress in the articular cartilage. This, and the thickness of the cartilage, also had the largest effect on the stresses in all the other tissues with the exception of the subchondral bone plate, in which stresses were dominated by its own stiffness. The stiffness of the underlying subchondral bone had no effect on the stresses generated in the cartilage. This study shows how stresses in the various tissues are affected by changes in their mechanical properties and thicknesses. It also demonstrates the benefits of a structured, systematic approach to investigating parameter variation in finite element models. PMID- 14558647 TI - Finite element prediction of endosteal and periosteal bone remodelling in the turkey ulna: effect of remodelling signal and dead-zone definition. AB - Bone remodelling is the adaptation of bone mass in response to localized changes in loading conditions. The nature of the mechanical signal governing remodelling, however, remains the subject of continued investigation. The aims of this study were to use an iterative finite element (FE) bone remodelling technique to explore the effect of different remodelling signals in the prediction of bone remodelling behaviour. A finite element model of the turkey ulna, following that of Brown et al., was analysed using the ABAQUS package. The model was validated against the static predictions of the Brown et al. study. A bone remodelling technique, based on swelling algorithms given by Taylor and Clift, was then applied to predict the dramatic change in loading conditions imposed. The resulting changes in FE mid-shaft bone geometry were compared with the remodelling observed experimentally and showed good agreement. The tensile principal stress was found to be the best remodelling signal under the imposed conditions. Localized sensitivities in the remodelling patterns were found, however, and the definition of the dead zone was modified as a result. Remodelling with the new dead-zone definition showed that both the tensile principal stress and the tensile principal strain produced the remodelling patterns that agreed most closely with experiment. PMID- 14558648 TI - Nanoindentation properties of compression-moulded ultra-high molecular weight polyethylene. AB - This paper investigates the elastic modulus and hardness of untreated and treated compression-moulded ultra-high molecular weight polyethylene (UHMWPE) tibial inserts of a total knee replacement (TKR) prosthesis. Investigations were carried out at a nanoscale using a Nanoindenter at penetration depths of 100,250 and 500 nm. The nanomechanical properties of surface and subsurface layers of the compression-moulded tibial inserts were studied using the untreated UHMWPE. The nanomechanical properties of intermediate and core layers of the compression moulded tibial insert were studied using the cryoultrasectioned and etched UHMWPE treated samples. The cryoultrasectioning temperature (-150 degrees C) of the samples was below the glass transition temperature, Tg (-122 +/- 2 degrees C ), of UHMWPE. The measurement of the mechanical response of crystalline regions within the nanostructure of UHMWPE was accomplished by removing the amorphous regions using a time-varying permanganic-etching technique. The percentage crystallinity of UHMWPE was measured using differential scanning calorimetry (DSC) and the Tg of UHMWPE was determined by dynamic mechanical analysis (DMA). Atomic force microscopy (AFM) was used to assess the effect of surface preparation on the samples average surface roughness, Ra. In this study, it was demonstrated that the untreated UHMWPE samples had a significantly lower (p < 0.0001) elastic modulus and hardness relative to treated UHMWPE cryoultrasectioned and etched samples at all penetration depths. No significant difference (p > 0.05) in elastic modulus and hardness between the cryoultrasectioned and etched samples was observed. These results suggest that the surface nanomechanical response of an UHMWPE insert in a total joint replacement (TJR) prosthesis is significantly lower compared with the bulk of the material. Additionally, it was concluded that the nanomechanical response of material with higher percentage crystallinity (67 per cent) was predominantly determined by the crystalline regions within the semi-crystalline UHMWPE nanostructure. PMID- 14558649 TI - Yielding of the clamped-wire system in the Ilizarov external fixator. AB - This study demonstrates that the clamped-wire system used to suspend bones within an Ilizarov external fixator yields when the perpendicular load exceeds 50 N per wire. Cyclic loading was applied to tensioned wires clamped within an Ilizarov ring component, with steadily increasing load amplitude. Wires were tested at four initial tension settings. The amount of energy lost within the clamped-wire system per load cycle was calculated for every test. The results showed that there was a consistent trend to increasing non-recoverable energy loss per load cycle when peak loads exceed 50 N for all initial tension settings. A finite element (FE) model replicating the experimental conditions was performed to investigate the levels of stress within the loaded wires. The FE model analyses showed that high stresses were generated in the wires close to the clamping sites, and that the stress levels could reasonably be expected to exceed the material yield stress when loaded to about 55 N, for all initial tension settings. The results show that material yield, accompanied by some wire slippage through the clamps, is responsible for system yield, in agreement with previous studies. Although the initial wire tension has an appreciable effect on the wire stiffness, it did not affect the elastic load range of the clamped-wire system. To prevent yield of the clamped-wire system in practice, the fixator should be assembled with sufficient wires to ensure that the load transmitted to each wire by the patient does not exceed 50 N. PMID- 14558650 TI - Taguchi's technique: an effective method for improving X-ray medical radiographic screen performance. AB - Taguchi's technique is a helpful tool to achieve experimental optimization of a large number of decision variables with a small number of off-line experiments. The technique appears to be an ideal tool for improving the performance of X-ray medical radiographic screens under a noise source. Currently there are very many guides available for improving the efficiency of X-ray medical radiographic screens. These guides can be refined using a second-stage parameter optimization. based on Taguchi's technique, selecting the optimum levels of controllable X-ray radiographic screen factors. A real example of the proposed technique is presented giving certain performance criteria. The present research proposes the reinforcement of X-ray radiography by Taguchi's technique as a novel hardware mechanism. PMID- 14558651 TI - Reciprocal walking orthosis with composite material body brace: initial development. AB - Reciprocal walking orthoses are routinely used by thoracic lesion patients for ambulation using crutches. A primary reason for their prescription is to provide therapeutic benefit and improved independence. To achieve this, maximum efficiency of walking and acceptance of the device is necessary to promote long term compliance. Lateral rigidity in the orthosis influences walking efficiency, but the structural properties of conventional techniques for producing a sufficiently rigid body brace makes them unattractive. Currently patients and clinicians are forced to choose between greater efficiency or cosmesis of the orthosis. Composite materials have the potential to permit the required rigidity in a structure that is less obtrusive. However, their material properties could lead to unsafe forms of failure unless suitable manufacturing methods are devised. It is therefore inappropriate to supply prototypes to patients for field evaluation until laboratory investigation of innovative production methods has ensured that the orthosis is safe. A production technique has been devised that is ostensibly suitable. Prototype body braces have been tested and have been shown to have improved structural properties and safe failure modes. A test programme implemented on a complete concept orthosis has confirmed that improved lateral rigidity can be achieved with a less obtrusive body brace, and that it will behave safely for long enough to permit field evaluation. PMID- 14558652 TI - Various issues relating to computational fluid dynamics simulations of carotid bifurcation flow based on models reconstructed from three-dimensional ultrasound images. AB - Computational fluid dynamics (CFD) flow simulation techniques have the potential to enhance understanding of how haemodynamic factors are involved in atherosclerosis. Recently, three-dimensional ultrasound has emerged as an alternative to other three-dimensional imaging techniques, such as magnetic resonance angiography (MRA). The method can be used to generate accurate vascular geometry suitable for CFD simulations and can be coupled with Doppler ultrasound to provide physiologically realistic flow boundary conditions. However, there are various ways to utilize the flow data acquired, possibly leading to different results regarding both flow and wall shear stress patterns. A disadvantage of three-dimensional ultrasound for imaging the carotid bifurcation has been established as being the scanning limitation of the jawbone position. This may make artificial extensions of the internal and/or external carotid arteries necessary, which in turn may influence the predicted flow patterns. Flow simulations were carried out for three outflow calculation schemes as well as four geometries with different extensions to the carotid daughter vessels. It was found that variation of flow patterns was more strongly influenced by the outflow conditions than by the extensions of the daughter vessels. Consequently, it is recommended that for future CFD simulations of carotid flow using three dimensional ultrasound data, the outflow boundary conditions should rely on the most accurate measurement available, and flow data recorded in the common and internal carotid are considered more reliable than data from the external carotid. Even though the extended lengths of the daughter vessels have insignificant effects on the predicted haemodynamic parameters, it would be a safer option to extend the internal carotid by approximately three times the diameter of the common carotid artery. PMID- 14558653 TI - Moment generation in wheelchair propulsion. AB - Wheelchair propulsion is a man machine interaction in which chair design and fit affect the relative positions and orientations of the upper extremity relative to the handrim and wheel axle. To understand these relationships better, experimental data were collected in five hand positions from five subjects exerting maximal effort to propel an instrumented wheelchair with its wheel in a locked position. The results of experiments revealed that the progression moment was greater at both initial and terminal propulsion positions and smaller in the mid-propulsion position. The vertical and horizontal force components were directed radially away from the wheel axle posterior to the dead centre position and radially towards the wheel axle anterior to top dead centre. Subsequently, a subject-specific quasi-static model of the upper extremity which maximized wheel progression moment was developed to augment our understanding of experimental measures. Model-predicted trends in progression moments and hand force direction were similar to experiment. Model predictions revealed that the optimal progression moment generation could potentially be affected by an individual's anthropometric parameters, joint strengths and also the direction of force applied by the hand on the handrim. Through wheelchair fitting and training of wheelchair users, it may be possible to improve propulsion technique. PMID- 14558654 TI - Genetic interactions between FLM and other flowering-time genes in Arabidopsis thaliana. AB - FLOWERING LOCUS M (FLM) is a MADS-domain gene that acts as an inhibitor of flowering in Arabidopsis. Here we describe the genetic interaction of FLM with genes in the photoperiod and autonomous flowering pathways. Although the sequence of FLM is most similar to that of FLC, FLM and FLC interact with different flowering pathways. It has been previously shown that flc lesions suppress the late-flowering phenotype of FRI-containing lines and autonomous-pathway mutants. However, flm lesions suppress the late-flowering phenotype of photoperiod-pathway mutants but not that of FRI-containing lines or autonomous-pathway mutants. Another MADS-domain flowering repressor with a mutant phenotype similar to FLM is SVP. The late-flowering phenotype of FLM over-expression is suppressed by the svp mutation, and an svp flm double mutant behaves like the single mutants. Thus FLM and SVP are in the same flowering pathway which interacts with the photoperiod pathway. PMID- 14558655 TI - Abundance of plastid DNA insertions in nuclear genomes of rice and Arabidopsis. AB - Pairwise comparison of whole plastid and draft nuclear genomic sequences of Arabidopsis thaliana and Oryza sativa L. ssp. indica shows that rice nuclear genomic sequences contain homologs of plastid DNA covering about 94 kb (83%) of plastid genome and including one or more full-length intact (without mutations resulting in premature stop codons) homologues of 26 known protein-coding (KPC) plastid genes. By contrast, only about 20 kb (16%) of chloroplast DNA, including a single intact plastid-derived KPC gene, is presented in the nucleus of A. thaliana. Sixteen rice plastid genes have at least one nuclear copy without any mutation or with only synonymous substitutions. Nuclear copies for other ten plastid genes contain both synonymous and non-synonymous substitutions. Multiple ESTs for 25 out of 26 KPC genes were also found, as well as putative promoters for some of them. The study of substitutions pattern shows that some of nuclear homologues of plastid genes may be functional and/or are under the pressure of the positive natural selection. The similar comparative analysis performed on rice chromosome 1 revealed 27 contigs containing plastid-derived sequences, totalling about 84 kb and covering two thirds of chloroplast DNA, with the intact nuclear copies of 26 different KPC genes. One of these contigs, AP003280, includes almost 57 kb (45%) of chloroplast genome with the intact copies of 22 KPC genes. At the same time, we observed that relative locations of homologues in plastid DNA and the nuclear genome are significantly different. PMID- 14558656 TI - Novel gene expression profiles define the metabolic and physiological processes characteristic of wood and its extractive formation in a hardwood tree species, Robinia pseudoacacia. AB - Wood is of critical importance to humans as a primary feedstock for biofuel, fiber, solid wood products, and various natural compounds including pharmaceuticals. The trunk wood of most tree species has two distinctly different regions: sapwood and heartwood. In addition to the major constituents, wood contains extraneous chemicals that can be removed by extraction with various solvents. The composition and the content of the extractives vary depending on such factors as, species, growth conditions, and time of year when the tree is cut. Despite the great commercial and keen scientific interest, little is known about the tree-specific biology of the formation of heartwood and its extractives. In order to gain insight on the molecular regulations of heartwood and its extractive formation, we carried out global examination of gene expression profiles across the trunk wood of black locust (Robinia pseudoacacia L.) trees. Of the 2,915 expressed sequenced tags (ESTs) that were generated and analyzed in the current study, 55.3% showed no match to known sequences. Cluster analysis of the ESTs identified a total of 2278 unigene sets, which were used to construct cDNA microarrays. Microarray hybridization analyses were then performed to survey the changes in gene expression profiles of trunk wood. The gene expression profiles of wood formation differ according to the region of trunk wood sampled, with highly expressed genes defining the metabolic and physiological processes characteristic of each region. For example, the gene encoding sugar transport had the highest expression in the sapwood, while the structural genes for flavonoid biosynthesis were up-regulated in the sapwood heartwood transition zone. This analysis also established the expression patterns of 341 previously unknown genes. PMID- 14558657 TI - Functional analysis of the rice AP3 homologue OsMADS16 by RNA interference. AB - The rice OsMADS16 gene is phylogenetically related to the angiosperm B-function MADS-box genes. To investigate if OsMADS16 functions as an AP3/DEF orthologue to regulate the development of lodicules and stamens in rice, we isolated its genomic sequences and characterized its functions in planta by RNA interference. The genomic sequence of the OsMADS16 gene shows that it shares high similarity in genomic structure and the deduced amino acid sequence with the maize B-class gene, Si1. Transgenic lines from the introduced gene expressing double-stranded RNA with the OsMADS16 cDNA fragment were male-sterile and displayed alternations of lodicules and stamens, occasionally depressed palea and overgrown glume. The two lodicules were converted into four palea/lemma-like organs and some stamens into carpels. Further investigations of the transcription of OsMADS16 gene in these transgenic lines by RT-PCR revealed that its transcript was significantly reduced. Transcription of a rice PI homologous gene, OsMADS4, was also reduced remarkably in the transgenic plants. Our results demonstrate that OsMADS16 is an AP3/DEF orthologue to specify the identities of lodicules and stamens in rice flower and also support that OsMADS4 is a PI orthologue. In addition, these results suggest that RNA interference is a useful tool for functional genomics in rice. PMID- 14558658 TI - Expression of the cation transporter McHKT1 in a halophyte. AB - From the ice plant, Mesembryanthemum crystallinum, McHKT1 was isolated encoding a protein 41-61% identical to other plant HKT1-like sequences previously described as potassium or sodium/potassium transporters. McHKT1 acts as a potassium transporter in yeast with specificity similar to that of wheat HKT1. In Xenopus oocytes it transports cations with a specificity Rb+ > Cs+ > [K+ = Na+ = Li+]. McHKT1 is exclusively localized to the plasma membrane. The isoform isolated is most highly expressed in leaves and is present in stems, flowers and seed pods but absent from the root where, according to immunological data, a second isoform exists which does not cross-hybridize with the leaf form in RNA blots at high stringency. McHKT1 transcript amounts increase during the first 6-10 h of stress and then decline to pre-stress levels with kinetics reminiscent of the initial influx of sodium into this halophyte. Immunocytological localization showed strong signals in the leaf vasculature and surrounding mesophyll cells but low intensity signals are also detected in other cell types. In roots, McHKT is mainly confined to endodermis and stele. Possible functions of McHKT1 in ion homeostasis in the halophytic ice plant are discussed. PMID- 14558660 TI - Generation of Arabidopsis protein chips for antibody and serum screening. AB - Protein array technology has emerged as a new tool to enable ordered screening of proteins for expression and molecular interactions in high throughput. Besides classical solid-phase substrates, such as micro-titre plates and membrane filters, protein arrays have recently been devised with chip-sized supports. Several applications on protein chips have been described, but to our knowledge no studies using plant protein chips were published so far. The aim of this study was to generate Arabidopsis protein chips and to demonstrate the feasibility of the protein chip technology for the investigation of antigen-antibody interactions. Therefore, Arabidopsis cDNAs encoding 95 different proteins were cloned into a GATEWAY-compatible Escherichia coli expression vector. RGS-His6 tagged recombinant proteins were purified in high throughput and robotically arrayed onto glass slides coated either with a nitrocellulose based polymer (FAST slides) or polyacrylamide (PAA slides). Using an anti-RGS-His6 antibody all proteins were detected on the chips. The detection limit was ca. 2-3.6 fmol per spot on FAST slides or 0.1-1.8 fmol per spot on PAA slides. The Arabidopsis protein chips were used for the characterisation of monoclonal antibodies or polyclonal sera. We were able to show that a monoclonal anti-TCP1 antibody and anti-MYB6 and anti-DOF11 sera bound specifically to their respective antigens and did not cross-react with the other 94 proteins including other DOF and MYB transcription factors on the chips. To enable screening of antibodies or other interacting molecules against thousands of Arabidopsis proteins in future, we generated an ordered cDNA expression library and started with high-throughput cloning of full-length cDNAs with GATEWAY technology. PMID- 14558659 TI - Light and circadian regulation in the expression of LHY and Lhcb genes in Phaseolus vulgaris. AB - In order to understand some aspects of the circadian clock function in Phaseolus vulgaris, we analyzed the temporal transcript profile of Lhcb genes, typical clock reporters in plants, and that of PvLHY, an orthologue of Arabidopsis thaliana LHY which is a putative transcription factor of Lhcb genes. Under different light regimes, Lhcb and PvLHY exhibit a clear circadian pattern of expression. Moreover, the rhythm of Lhcb genes appears to be tightly coupled to that of PvLHY with the latter having a slightly earlier phase. This supports the idea that the oscillating capacity of PvLHY may be one of the causes of the rhythmic expression of Lhcb genes in bean. In addition to their circadian regulation, Lhcb and PvLHY are induced by light with similar and relatively slow induction kinetics. Moreover, this light induction is gated by the circadian oscillator: minimal responses occur at times around peaks of the pre-existing rhythm, while maximal ones occur at troughs of the pre-existing rhythm. This pattern of gating is opposite to that observed in Arabidopsis. The failure to block the light induction pathways at pre-existing troughs appears to have a detrimental effect to the subsequent circadian rhythmicity. Briefly, the overall regulation of PvLHY and Lhcb genes by light and the circadian clock reveals different strategies between Phaseolus and Arabidopsis in the adaptation to photoperiodic conditions. PMID- 14558662 TI - Molecular characterization and spatial expression of the sunflower ABP1 gene. AB - We have used RT-PCR and low-stringency cDNA library screening to isolate the coding sequence of the sunflower auxin-binding protein (ABP1). All the clones analysed contained the same nucleotide sequence, suggesting that ABP1 is encoded by a single-copy gene in sunflower. The deduced amino acid sequence shows a high degree of similarity with ABP1 proteins from other plant species. Most remarkably, the sunflower protein lacks two cysteine residues present in all other plant ABPs known to date and shown to be involved in a disulfide bridge in the maize protein. Genomic Southern hybridization data support the existence of a single copy of the ABP1 gene in the sunflower genome. Northern hybridization corroborated earlier observations indicating that the steady-state level of ABP1 transcript is higher in actively dividing and growing organs than in the rest of the plant: it is more abundant in the shoot apex, floral buds and immature embryos than in mature leaves, stem, roots and ray flowers. To characterize the tissular ABP1 transcript distribution in sunflower, various organ sections were analysed upon in situ hybridization. Localized accumulation of the ABP1 transcript suggests that its spatial expression is highly regulated at the tissue level. In addition, the transcript preferentially accumulates in tissues having a high rate of cellular division, such as shoot and root apical meristems, leaf primordia and pro-vascular tissues. The ABP1 expression pattern was also studied at a temporal scale during lateral root formation. Real time PCR showed an elevation of the steady state level of the ABP1 transcript in root axes after 36 h of seed germination. In situ hybridization revealed that this global increase is the result of local accumulation of the ABP1 transcript in lateral root primordia, which are known to develop under auxin action. The possibility that a high ABP1 expression level correlates with a high cellular sensitivity to auxin is discussed. PMID- 14558661 TI - Regulation of CDPK isoforms during tuber development. AB - CDPK activities present during tuber development were analysed. A high CDPK activity was detected in the soluble fraction of early stolons and a lower one was detected in soluble and particulate fractions of induced stolons. The early and late CDPK activities displayed diverse specificity for in vitro substrates and different subcellular distribution. Western blot analysis revealed two CDPKs of 55 and 60 kDa that follow a precise spatial and temporal profile of expression. The 55 kDa protein was only detected in early-elongating stolons and the 60 kDa one was induced upon stolon swelling, correlating with early and late CDPK activities. A new member of the potato CDPK family, StCDPK3, was identified from a stolon cDNA library. Gene specific RT-PCR demonstrated that this gene is only expressed in early stolons, while the previously identified StCDPK1 is expressed upon stolon swelling. This expression profile suggests that StCDPK3 could correspond to the 55 kDa isoform while StCDPK1 could encode the 60 kDa isoform present in swelling stolons. StCDPK1 has myristoylation and palmitoylation consensus possibly involved in its dual intracellular localization. Transient expression studies with wild-type and mutated forms of StCDPK1 fused to GFP were used to show that subcellular localization of this isoform is controlled by myristoylation and palmitoylation. Altogether, our data suggest that sequential activation of StCDPK3 and StCDPK1 and the subcellular localisation of StCDPK1 might be critical regulatory steps of calcium signalling during potato tuber development. PMID- 14558663 TI - Cloning and characterization of the durable tomato mosaic virus resistance gene Tm-2(2) from Lycopersicon esculentum. AB - In tomato, infections by tomato mosaic virus are controlled by durable Tm-2(2) resistance. In order to gain insight into the processes underlying disease resistance and its durability, we cloned and analysed the Tm-2(2) resistance gene and the susceptible allele, tm-2. The Tm-2(20 gene was isolated by transposon tagging using a screen in which plants with a destroyed Tm-2(2) gene survive. The Tm-2(2) locus consists of a single gene that encodes an 861 amino acid polypeptide, which belongs to the CC-NBS-LRR class of resistance proteins. The putative tm-2 allele was cloned from susceptible tomato lines via PCR with primers based on the Tm-2(2) sequence. Interestingly, the tm-2 gene has an open reading frame that is comparable to the Tm-2(2) allele. Between the tm-2 and the Tm-2(2) polypeptide 38 amino acid differences are present of which 26 are located in the second half of the LRR-domain. Susceptible tomato plants, which were transformed with the Tm-2(2) gene, displayed resistance against ToMV infection. In addition, virus specificity, displayed by the Tm-2(2) resistance was conserved in these transgenic lines. To explain the durability of this resistance, it is proposed that the Tm-2(2)-encoded resistance is aimed at the Achilles' heel of the virus. PMID- 14558664 TI - An antirrhinum ternary complex factor specifically interacts with C-function and SEPALLATA-like MADS-box factors. AB - The development of floral reproductive organs requires the activity of plant MADS box transcription factors (MBFs) belonging to the C function. The C function can only operate within a floral context, specified by MBFs belonging to the SEPALLATA class of proteins. Here we describe the specific interaction between a novel protein, MIP1, and C-function and SEPALLATA (SEP)-like MBFs. MIP1 is the first member of a new class of proteins unique to plants. None of the family members have yet been assigned a function. Motif searches reveal a leucine zipper domain within a conserved N-terminal region of MIP1. The leucine zipper lies within a region sufficient for interaction with plant MBFs. MIP1 interacts with a domain of plant MBFs that is analogous to the domain of animal and yeast MBFs involved in ternary complex formation. The MIP1 protein is predicted to localise to the nucleus and activates yeast reporter genes in vivo. MIP1 is expressed in the fourth whorl of the flower, in an overlapping temporal and spatial expression pattern with the C-function and SEP-like genes. Taken together, this suggests that MIP1 acts as a ternary complex factor specifically with C-function and SEP like MBFs. PMID- 14558665 TI - Molecular characterization of a nuclear topoisomerase II from Nicotiana tabacum that functionally complements a temperature-sensitive topoisomerase II yeast mutant. AB - We have successfully expressed enzymatically active plant topoisomerase II in Escherichia coli for the first time, which has enabled its biochemical characterization. Using a PCR-based strategy, we obtained a full-length cDNA and the corresponding genomic clone of tobacco topoisomerase II. The genomic clone has 18 exons interrupted by 17 introns. Most of the 5' and 3' splice junctions follow the typical canonical consensus dinucleotide sequence GU-AG present in other plant introns. The position of introns and phasing with respect to primary amino acid sequence in tobacco TopII and Arabidopsis TopII are highly conserved, suggesting that the two genes are evolved from the common ancestral type II topoisomerase gene. The cDNA encodes a polypeptide of 1482 amino acids. The primary amino acid sequence shows a striking sequence similarity, preserving all the structural domains that are conserved among eukaryotic type II topoisomerases in an identical spatial order. We have expressed the full-length polypeptide in E. coli and purified the recombinant protein to homogeneity. The full-length polypeptide relaxed supercoiled DNA and decatenated the catenated DNA in a Mg(2+) and ATP-dependent manner, and this activity was inhibited by 4'-(9 acridinylamino)-3'-methoxymethanesulfonanilide (m-AMSA). The immunofluorescence and confocal microscopic studies, with antibodies developed against the N terminal region of tobacco recombinant topoisomerase II, established the nuclear localization of topoisomerase II in tobacco BY2 cells. The regulated expression of tobacco topoisomerase II gene under the GAL1 promoter functionally complemented a temperature-sensitive TopII(ts) yeast mutant. PMID- 14558666 TI - A plasma membrane zinc transporter from Medicago truncatula is up-regulated in roots by Zn fertilization, yet down-regulated by arbuscular mycorrhizal colonization. AB - Here we present a Zn transporter cDNA named MtZIP2 from the model legume Medicago truncatula. MtZIP2 encodes a putative 37 kDa protein with 8-membrane spanning domains and has moderate amino acid identity with the Arabidopsis thaliana Zn transporter AtZIP2p. MtZIP2 complemented a Zn-uptake mutant of yeast implying that the protein encoded by this gene can transport Zn across the yeast's plasma membrane. The product of a MtZIP2-GFP fusion construct introduced into onion cells by particle bombardment likewise localized to the plasma membrane. The MtZIP2 gene was expressed in roots and stems, but not in leaves of M. truncatula and, in contrast to all other plant Zn transporters characterized thus far, MtZIP2 was up-regulated in roots by Zn fertilization. Expression was highest in roots exposed to a toxic level of Zn. MtZIP2 expression was also examined in the roots of M. truncatula when colonized by the obligate plant symbiont, arbuscular mycorrhizal (AM) fungi, since AM fungi are renowned for their ability to supply plants with mineral nutrients, including Zn. Expression was down-regulated in the roots of the mycorrhizal plants and was associated with a reduced level of Zn within the host plant tissues. PMID- 14558667 TI - Protein sorting and expression of a unique soybean cotyledon protein, GmSBP, destined for the protein storage vacuole. AB - The initial biochemical characterization of the soybean sucrose-binding protein, GmSBP, within our lab and others produced several incongruous characteristics that required a re-characterization of GmSBP via sequence homology, cell biology, immunolocalization, and semi-quantitative analysis. The GmSBP proteins share amino acid sequence homology as well as putative structural homology with globulin-like seed storage proteins. A comparison to the major soybean seed storage proteins, glycinin and beta-conglycinin established several storage protein-like characteristics for GmSBP. All three proteins were present in a prevacuolar compartment and protein storage vacuole. All three proteins increased in expression during seed development and are remobilized during germination. Quantitatively, the relative concentrations of GmSBP, beta-conglycinin (alpha/alpha' subunits), and glycinin (acidic subunits) indicated that GmSBP contributes 19-fold less to the stored nitrogen. The quantitative differences between GmSBP and glycinin may be attributed to the unconserved order and spacing of cis-acting regulatory elements present within the promoter regions. Ultimately, GmSBP is transported to the mature protein storage vacuole. The biological function of GmSBP within the protein storage vacuole remains uncertain, but its localization is a remnant of its evolutionary link to a globulin-like or vicilin-like ancestor that gave rise to the 7S family of storage proteins. PMID- 14558668 TI - Genomic instability in pituitary adenomas. AB - Pituitary adenomas most commonly are identified as small, incidental microadenomas. They however may progress to macroadenoma forming intra and later suprasellar tumors which in about 1/3 of cases invade surrounding structures at the time of diagnosis. Mechanism of pituitary tumorigenesis remains still elusive. Because the value of karyotyping is limited by the technical problems related to cytogenetic methods, we studied the spectrum of chromosomal imbalances associated with pituitary adenoma using comparative genomic hybridization (CGH). Copy number aberrations on all 22 autosomes were evaluated by CGH using advanced computer software. In total, fifteen patients were included in the study of 9 non invasive, 4 invasive and two recurrent adenomas. The mean age of the patients were 48 years ranging from 36 to 68 years. Five tumors showed hormonal activity. The histogram of all 15 cases representing the DNA imbalances as an incidence curve along each chromosome showed losses particularly for chromosomes 1p, 2q, 4, 5, 6, 11q, 12q, 13q and 18q as well as overrepresentation on 9q, 16p, 17p, 19, 20q. Functioning adenomas carried more imbalances than non-functioning, specifically deletions on chromosome 4 and 18q as well as overrepresentations of chromosomes 17 and 19. Invasive adenomas carried more overrepresentations at 1p34 than non-invasive tumors. Recurrent adenomas harbored more alterations than primary tumors, particularly DNA gains. The primary data is accessible at our CGH online tumor database at http://amba.charite.de/cgh. Reviewing the existing literature on the genetics of pituitary adenoma and discussing our results in this context, we hope that our study will contribute to the knowledge of this neoplasm. PMID- 14558669 TI - Significance of absent prohormone convertase 1/3 in inducing clinically silent corticotroph pituitary adenoma of subtype I--immunohistochemical study. AB - Biologically inactive ACTH-producing pituitary adenoma is known as clinically silent corticotroph adenoma. To search for the mechanism causing clinically silent corticotroph adenoma, we immunohistochemically examined the expression of prohormone convertase 1/3 (PC1/3) in this type of adenoma and compared our results with those obtained for Cushing's disease. All of the Cushing's disease specimens exhibited strongly positive PC1/3 exhibition. On the contrary, the expression of PC1/3 was very weak in the clinically silent corticotroph adenoma specimens. The absence of PC1/3 in clinically silent corticotroph adenoma indicates that silent corticotroph adenomas arise in a different cell type sharing the prohormone pro-opiomelanocortin (POMC), but processing it differently, accounting for the lack of clinical symptoms due to ACTH excess. PMID- 14558670 TI - Management of recurrent pituitary cysts with pituitary-nasal drain. AB - Pituitary cysts are usually craniopharyngiomas or remnants of Rathke's cleft. Histologically craniopharyngiomas are lined by stratified squamous epithelium whereas Rathke's cleft cysts are lined by cuboidal or columnar epithelium. The management of pituitary cysts remains difficult because of the risks associated with attempts at complete removal and the chances of recurrence after inadequate resection. We describe 3 cases of cystic craniopharyngiomas and one case of Rathke's cleft cyst which recurred after surgery and were subsequently managed successfully with pituitary-nasal drains. PMID- 14558671 TI - The expression of the F-box protein Skp2 is negatively associated with p27 expression in human pituitary tumors. AB - The CDK inhibitor p27 plays a pivotal role in controlling cell proliferation during development, and has been implicated in tumorigenesis. Previous studies have demonstrated changes in p27 protein expression, but not in mRNA levels, in human pituitary tumors. It seems probable that the fall in p27 is due to increased degradation through the ubiquitin-proteasome pathway. Skp2 (S-phase kinase-interacting protein) is a specific F-box protein that allows the recognition and binding of phosphorylated p27 to the ubiquitin complex. The aim of our study was thus to investigate the possible role of Skp2 in pituitary tumorigenesis. A total of 59 human pituitary samples, 7 normal and 52 adenomas, were assessed for transcriptional expression of Skp2; 51 pituitary samples were assessed for protein expression. Real-time RT-PCR was performed on cDNA of reverse-transcribed mRNA for relative quantification of the Skp2 transcript. Immunostaining was performed using mouse monoclonal anti-Skp2 antibody. Skp2 mRNA and protein was detectable in every sample studied. Our results showed no significant difference between the pituitary tumors and normal pituitary tissue in Skp2 mRNA or nuclear protein expression. Individual tumor types had similar mRNA expression and variable protein expression. However, samples with high p27 protein expression showed significantly less Skp2 expression than samples with low p27 staining. Our data suggest that increased p27 degradation through the ubiquitin-proteasome pathway could be regulated in pituitary tumors by changes in Skp2 expression, although other factors probably also play a role. PMID- 14558672 TI - Medical therapy of macroprolactinomas in males: I. Prevalence of hypopituitarism at diagnosis. II. Proportion of cases exhibiting recovery of pituitary function. AB - Hyperprolactinaemia frequently causes secondary hypogonadism through central suppression of gonadotropin secretion. Macroprolactinomas (> 1 cm diameter) are more common in males and may additionally cause more generalised hypopituitarism. Recovery of the thyrotropic and/or corticotropic axes is well described following selective adenomectomy, but remains poorly defined in relation to medical (dopamine-agonist) therapy of macroprolactinomas. We therefore performed a retrospective examination of case records of male patients who had received medical therapy alone for macroprolactinoma between 1980-2001 (n = 35) and in whom tumor shrinkage was documented by interval pituitary imaging (reported throughout by a single neuroradiologist). Mean prolactin level at baseline was 59,932 mU/L (median 31,400; range 3,215-332,000); mean period of follow up was 4.2 years (median 2.6; range: 1.0-15). Defects of the following axes were evident at diagnosis: LH/FSH-testosterone (n = 27; 77%), TSH-T4 (n = 14; 41%-not including one case with pre-existing 1 degress hypothyroidism), ACTH-cortisol (n = 8; 23%). Overall, 14 men (40%) were deficient in 1 axis, seven (20%) in 2 axes and seven (20%) in 3 axes. Growth hormone secretory status was not systematically evaluated. In all but 6 patients, prolactin levels fell to normal or near-normal levels (mean 764 mU/L; median 260; range: < 10-4,833). Of the patients in whom adequate reassessment had been performed, thyrotroph function recovered in 4/9, corticotroph function in 4/6 and gonadotroph function in 16/26 cases. In four cases (11%) previously described, development of visual impairment as a result of the chiasmal traction syndrome necessitated a dose reduction in medical therapy to allow a degree of controlled tumor re-expansion. The prevalence at diagnosis of TSH and ACTH deficiency in men with macroprolactinomas was 41% and 23%, respectively. Among eight patients with insufficiency of TSH and/or ACTH secretion who underwent complete interval reassessment over several years of treatment, recovery of at least one axis occurred in six cases (75%). This study highlights the importance of screening ACTH- and/or TSH-deficient men during dopamine agonist therapy in order to identify cases where hypopituitarism has resolved. PMID- 14558673 TI - Gender-related differences in growth hormone-releasing pituitary adenomas. A clinicopathological study. AB - BACKGROUND/AIMS: Pituitary adenomas are the third most common primary intracranial neoplasm, after astrocytomas and meningiomas, and about 30% of them secrete growth hormone (GH). Other subtypes of pituitary tumors are characterized by well-known gender-related differences, not only in clinical presentation and other biological characteristics but also in surgical outcome. For GH-releasing pituitary adenomas, however, detailed data on gender differences of postsurgical treatment are not available. PATIENTS AND METHODS: The patient charts of a series of 18 patients with acromegaly who met strict immunohistochemical and electron microscopic criteria and who underwent surgical resection of their tumors between January 1990 and June 1999 were retrospectively reviewed. RESULTS: There were eight women and ten men; the male-to-female-ratio was 1.3:1. The men and women were equal in age at surgery. Men demonstrated higher IGF-1 and smaller GH levels pre- and postoperatively, whereas the reduction in IGF-1 was more pronounced compared to women (58% vs. 27%). The overall outcome was better in women than in men. Mixed GH- and prolactin-secreting adenomas showed a worse outcome among all other histological subtypes. Mitose- and MIB-1 labeling index was increased in men compared to women. CONCLUSION: The clinical course and tumor biology of GH releasing pituitary adenomas appear to differ in women and men. Men demonstrated a shorter preoperative duration of symptoms, larger and more invasive tumors, and a worse clinical outcome. These findings suggest that therapy for GH-releasing adenomas should be more aggressive in men than in women. The gender-related differences in GH-releasing pituitary adenomas appear to have a basis in different biologic behavior, which warrants further investigation. PMID- 14558674 TI - Hyperprolactinemia in asymptomatic patients is related to high molecular weight posttranslational variants or glycosylated forms. AB - Circulating human Prolactin (PRL) exists in different variants related to posttranslational modifications, dimerization or association with other serum proteins. Compared to monomeric prolactin these variants usually have little or no biologic activity and include BigBig (BB PRL), Big (B PRL), and Glycosylated forms (G PRL). The aim of the present study was to assess levels of BB PRL, B PRL, little PRL (L PRL) and G PRL in hyperprolactinemic patients with no menstrual alterations or galactorrhea. L PRL, B PRL, and BB PRL were identified by gel filtration chromatography on Sephadex G-100; G PRL and NG PRL were identified by chromatography on Concanavalin A Sepharose. PRL was measured by IRMA DPC. Eleven women, aged 22-50 yrs, were studied for: breast dysplasia (1), controlled hypothyroidism (3), dysmenorrhea (3), microadenoma follow-up (2), and gynecological control (2). Pituitary MRI was normal in all but one patient, who had a microadenoma discovered by Magnetic Resonance Imaging. Six patients had normal L PRL levels, and their hyper PRL was due to excess BPRL or BB PRL. Five patients had increased L PRL levels, but excess G PRL. Patients harboring molecular PRL variants do not present the symptoms typical of the hyperprolactinemic syndrome. Furthermore in patients with clinically controlled prolactinomas the presence of PRL variants should be ruled out to avoid an unnecessary increase of dopamine agonist dosage. PMID- 14558675 TI - Falsely low serum prolactin in two cases of invasive macroprolactinoma. AB - The differential diagnosis of tumors at the base of the skull comprises meningiomas, neurinomas, gliomas, metastatic carcinomas, chordomas, epidermoids, and pituitary adenomas. About half of the pituitary adenomas are prolactinomas which are unique in a sense that medical therapy causes rapid tumor shrinkage and symptomatic improvement. We report on two patients in which the diagnosis of an invasive macroprolactinoma was masked by apparently low prolactin levels caused by a high-dose hook effect in the chemiluminometric assay. The first case a 49 year old male with impairment of hearing on the left side was presented in the Department of Otorhinolaryngology. A massive invasively growing tumor was demonstrated on a cranial MRI. Endocrine tests revealed normal pituitary function and normoprolactinemia. The patient underwent debulking surgery, occipitocervical fusion because of destruction of the first cervical vertebra and subsequent irradiation. The histopathological diagnosis was invasive prolactinoma. A repeat prolactin (PRL) sample, which was assayed using serial dilutions, revealed a real PRL level of 89,700 ng/ml. Dopamine agonist therapy was initiated under which PRL levels declined in parallel with tumor size. The second case a 40 year old male was presented with acute visual loss. Cranial MRI showed a large tumor at the base of the skull. Based on a transnasal biopsy, the preliminary diagnosis was a poorly differentiated carcinoma for which emergency irradiation was performed. Endocrine tests demonstrated partial hypopituitarism and moderate hyperprolactinemia. Hydrocortisone was substituted and dopamine agonist therapy was started because of moderate hyperprolactinemia. The final histopathological diagnosis was invasive prolactinoma. A repeat PRL sample assayed in serial dilution demonstrated an apparent rise in PRL with a maximum value of 6,460 ng/ml. Under dopamine agonist therapy, PRL declined to normal values, tumor size decreased and cranial nerve palsies disappeared. The apparently falsely low prolactin levels in the initial work-up of both patients were caused by a high dose hook effect in the PRL assay. Serial dilutions of serum PRL samples is, therefore, mandatory in the diagnostic work-up of patients with large invasive tumors at the base of the skull. This avoids unnecessary aggressive and dangerous treatment like surgery or radiotherapy in cases where pharmacological treatment may be the choice. PMID- 14558677 TI - Behavioral pediatrics, part II. PMID- 14558676 TI - An unusual presentation of Nelson's syndrome with apoplexy and subarachnoid hemorrhage. AB - A thirty-eight year-old lady with a history of bilateral adrenalectomy for Cushing's disease seven years previously, presented with sudden onset of severe headache, nausea, vomiting and loss of consciousness. She was somnolent and confused. She had neck stiffness, sixth nerve palsy and mydriasis on the left side. Computerized tomography (CT) and magnetic resonance imaging (MRI) studies revealed a non-homogeneous, grade IV D pituitary mass lesion associated with hemorrhage in the chiasmatic, interhemispheric, cerebellopontine, perimesencephalic cisterns and a hematoma within the frontal lobe. Angiography showed only bilateral elevation of horizontal segments of the anterior cerebral arteries. According to this angiographic evidence, it was presumed that the subarachnoid hemorrhage and the intracerebral hematoma were linked to pituitary adenoma apoplexy. ACTH level was 450 pg/ml. The hemorrhagic lesion with suprasellar extension was totally removed by left pterional craniotomy. Histological examination revealed a necrotic, ACTH-secreting pituitary adenoma. Even though apoplexy is a well known complication of pituitary adenomas, to our knowledge subarachnoid hemorrhage and intracerebral hematoma as a result of pituitary apoplexy in the context of Nelson's syndrome has not previously been reported. PMID- 14558678 TI - Violence: concepts of its impact on children and youth. AB - Although nations continue to remain involved in ongoing armed conflicts, the threat of direct exposure to violence for American children and youth is more likely to be from the interpersonal violence that occurs in homes, neighborhoods, and schools. Exposure to interpersonal violence has a very serious impact on most youth. Focusing on violence prevention remains a vital component of providing comprehensive health care for all youth. PMID- 14558679 TI - Disruptive behavioral disorders. AB - Disruptive behavior disorders are some of the most frequently encountered disorders in any setting. Most authorities agree that DBDs are a heterogeneous group of disorders with multiple etiological risk factors. DBD can manifest in a variety of ways and is unique for each individual. The multiple revisions for defining DBD, along with the complexities encountered in the diagnosis and treatment, create some very difficult cases. Fortunately, research continues to advance knowledge and understanding and provide newer options for treatment. Using the concepts and principles in a comprehensive multi-disciplinary team approach to the diagnosis and treatment continue to provide the best results. Because most offices and clinics are not able to offer this multitude and variety of professional services, making referrals becomes a necessity. It is important to emphasize the need of periodic reassessments to demonstrate and document progress and for modifications in the treatment plan. The American Academy of Child and Adolescent Psychiatry (AACAP) publishes a helpful summary article entitled "Practice Parameters for the Assessment and Treatment of Children and Adolescents with Conduct Disorder," and copies can be obtained by contacting the AACAP. PMID- 14558680 TI - Attention-deficit/hyperactivity disorder evaluation and diagnosis: a practical approach in office practice. AB - Attention-deficit/hyperactivity disorder is the most common neurobehavioral disorder of childhood, with most children maintaining symptoms of ADHD as adolescents and as adults. It is among the most common chronic conditions that primary care pediatricians see. Progress in brain imaging, genetics, neuropsychology, and molecular biology of ADHD reveals complex interactions between neurologic mechanisms, genetics, and environmental influences. Core behaviors seen in children with ADHD include hyperactivity, impulsivity, and inattention. The DSM-IV standard for diagnosis emphasized the importance of documenting that these behaviors occur at school and in the home, the chronicity of symptoms, and the connection between the behaviors and impairments in educational achievement or social development. The complex nature of this disorder is indicated by the observation that children and adolescents with ADHD have associated conditions, such as learning disorders, anxiety, oppositional behaviors, and depression. The AAP practice guideline on the evaluation and diagnosis of school-aged children with ADHD provides primary care pediatricians with a scientific foundation for assessing children with behavior and learning problems. Attention to accurate documentation of behaviors, coexisting learning disorders and mental health conditions, and the development of office practices that support the diagnostic process will ensure that children receive an appropriate and comprehensive assessment. PMID- 14558681 TI - Attention-deficit/hyperactivity disorder in children and adolescents: interventions for a complex costly clinical conundrum. AB - Management of a child or adolescent with attention-deficit/hyperactivity disorder (ADHD) is reviewed, including psychological and pharmacologic approaches. Psychological treatment includes psychotherapy, cognitive-behavior therapy, support groups, parent training, educator/teacher training, biofeedback, meditation, and social skills training. Medications are reviewed that research has revealed can improve the core symptomatology of a child or adolescent with ADHD. These medications include stimulants, antidepressants, alpha-2 agonists, and a norepinephrine reuptake inhibitor. Management of ADHD should include a multi-modal approach, involving appropriate educational interventions, appropriate psychological management of the patient (child or adolescent), and judicious use of medications. Parents, school officials, and clinicians must work together to help all children and adolescents with ADHD achieve their maximum potential. PMID- 14558682 TI - Depression in the child and adolescent. AB - Depression is a common disorder of childhood and adolescence for which effective interventions exist. Primary care physicians should be familiar with its essential features to ensure that their patients do not suffer needlessly from this potentially devastating, even fatal disorder. Clinical signs and symptoms of depression typically are not difficult to elicit, but patients and families may fail to volunteer them unless asked. A growing body of evidence exists to guide clinicians in their choice of treatment interventions. Treatment response to studied interventions is high, although recurrence is common. Consultation with a child and adolescent psychiatrist is advisable in complicated cases or severe presentations, especially when patients are suicidal or threatening to harm others. PMID- 14558683 TI - Anxiety disorders in the child and teen. AB - Several disorders have been reviewed (Table 1). Based upon review of the literature, an algorithm has been developed, supporting the initial use of cognitive behavioral therapy, followed by a psychopharmacology algorithm if treatment is not successful. In this algorithm, severely anxious patients initially may require psychopharmacologic treatment to be able to participate in cognitive behavioral treatment. Nonspecific measures of parent education, general support, and illness education to parents and patients are overarching principles. In this algorithm, the SSRIs are perceived to be first-line interventions, with tricyclic antidepressants and venlafaxine as second-line agents. Buspirone is considered a second- or third-line agent, as are the benzodiazepines. Table 2 reviews psychopharmacologic agents shown to be useful in the management of anxiety disorders in youth. Although much research remains to be done, there is evidence of efficacy of several interventions for anxiety disorders in children and adolescents. There is a need for a holistic and comprehensive management plan. Particular attention must be given to specific psychopharmacologic and psychotherapy needs, family matters, abuse issues, freedom from substance abuse, the use of peer support groups, and the encouragement of healthier lifestyle choices such as exercise. A rising number of well-done, large, placebo-controlled studies are providing increased support for medication and psychotherapy to inform evidence-based treatment. There is a need for teamwork and effective communication among team members in addressing pediatric and adolescent anxiety disorders. PMID- 14558684 TI - Eating disorders in children and adolescents. AB - Anorexia nervosa and bulimia nervosa are common disorders in the pediatric population. This article is designed to help the clinician develop a detailed understanding of these disorders as they affect children and adolescents. The etiology, diagnosis, complications, and treatment of these eating disorders are discussed. The special circumstances of diabetes mellitus, athletics, and the interface with the obese patient also are covered. The need to correctly identify a patient with the disorder, perform the appropriate testing, and organize the most appropriate treatment is required from virtually every primary care practitioner. PMID- 14558685 TI - Substance abuse in adolescents: a complex conundrum for the clinician. AB - Substance abuse remains a complex and pervasive conundrum for society and for clinicians seeking to improve the lives of their pediatric patients. Substance abuse is linked to the human instinct for pleasure at any cost and is fueled by enticing encouragement of the media teaching society to seek drug-induced pleasure without fear of negative consequences. Other complications are the limited education about psychoactive substances provided to youth and the health care profession pledged to serve them. Primary care clinicians must provide their adolescent patients with adequate screening and counseling about substance abuse. Treatment of the substance-abusing patient is often a combination of behavioral interventions (including family therapy), and, in limited situations, addiction specific medications. Research suggests that female drug addicts have a better outcome in female-only drug treatment programs. In addition, new drugs are being developed that target specific brain mechanisms involved in drug addiction; these drugs will have less toxicity and less abuse potential than illicit drugs such as cocaine. Vaccines are being developed that will block the effects of such drugs as cocaine and PCP. Medications developed for the treatment of depression and epilepsy will be a source of medications for the treatment of drug addiction. The study of endorphins and the neurobiology of stress and substance abuse promise to develop potent anti-addiction chemicals, greatly aiding in the war on drug abuse. PMID- 14558686 TI - Induced systemic resistance in Arabidopsis thaliana in response to root inoculation with Pseudomonas fluorescens CHA0. AB - Root inoculation of Arabidopsis thaliana ecotype Columbia with Pseudomonas fluorescens CHA0r partially protected leaves from the oomycete Peronospora parasitica. The molecular determinants of Pseudomonas fluorescens CHA0r for this induced systemic resistance (ISR) were investigated, using mutants derived from strain CHA0: CHA400 (pyoverdine deficient), CHA805 (exoprotease deficient), CHA77 (HCN deficient), CHA660 (pyoluteorin deficient), CHA631 (2,4 diacetylphloroglucinol [DAPG] deficient), and CHA89 (HCN, DAPG- and pyoluteorin deficient). Only mutations interfering with DAPG production led to a significant decrease in ISR to Peronospora parasitica. Thus, DAPG production in Pseudomonas fluorescens is required for the induction of ISR to Peronospora parasitica. DAPG is known for its antibiotic activity; however, our data indicate that one action of DAPG could be due to an effect on the physiology of the plant. DAPG at 10 to 100 microM applied to roots of Arabidopsis mimicked the ISR effect. CHA0r mediated ISR was also tested in various Arabidopsis mutants and transgenic plants: NahG (transgenic line degrading salicylic acid [SA]), sid2-1 (nonproducing SA), npr1-1 (non-expressing NPR1 protein), jar1-1 (insensitive to jasmonic acid and methyl jasmonic acid), ein2-1 (insensitive to ethylene), etr1-1 (insensitive to ethylene), eir1-1 (insensitive to ethylene in roots), and pad2-1 (phytoalexin deficient). Only jar1-1, eir1-1, and npr1-1 mutants were unable to undergo ISR. Sensitivity to jasmonic acid and functional NPR1 and EIR1 proteins were required for full expression of CHA0r-mediated ISR. The requirements for ISR observed in this study in Peronospora parasitica induced by Pseudomonas fluorescens CHA0r only partially overlap with those published so far for Peronospora parasitica, indicating a great degree of flexibility in the molecular processes leading to ISR. PMID- 14558687 TI - Fungal biology and agriculture: revisiting the field. AB - Plant pathology has made significant progress over the years, a process that involved overcoming a variety of conceptual and technological hurdles. Descriptive mycology and the advent of chemical plant-disease management have been followed by biochemical and physiological studies of fungi and their hosts. The later establishment of biochemical genetics along with the introduction of DNA-mediated transformation have set the stage for dissection of gene function and advances in our understanding of fungal cell biology and plant-fungus interactions. Currently, with the advent of high-throughput technologies, we have the capacity to acquire vast data sets that have direct relevance to the numerous subdisciplines within fungal biology and pathology. These data provide unique opportunities for basic research and for engineering solutions to important agricultural problems. However, we also are faced with the challenge of data organization and mining to analyze the relationships between fungal and plant genomes and to elucidate the physiological function of pertinent DNA sequences. We present our perspective of fungal biology and agriculture, including administrative and political challenges to plant protection research. PMID- 14558688 TI - Analysis of gene expression in two growth states of Xylella fastidiosa and its relationship with pathogenicity. AB - Xylella fastidiosa is a plant pathogen responsible for diseases of economically important crops. Although there is considerable disagreement about its mechanism of pathogenicity, blockage of the vessels is one of the most accepted hypotheses. Loss of virulence by this bacterium was observed after serial passages in axenic culture. To confirm the loss of pathogenicity of X. fastidiosa, the causing agent of citrus variegated chlorosis (CVC), freshly-isolated bacteria (first passage [FP] condition) as well as bacteria obtained after 46 passages in axenic culture (several passage [SP] condition) were inoculated into sweet orange and periwinkle plants. Using real time quantitative polymerase chain reaction, we verified that the colonization of FP cells was more efficient for both hosts. The sequence of the complete X. fastidiosa genome allowed the construction of a DNA microarray that was used to investigate the total changes in gene expression associated with the FP condition. Most genes found to be induced in the FP condition were associated with adhesion and probably with adaptation to the host environment. This report represents the first study of the transcriptome of this pathogen, which has recently gained more importance, since the genome of several strains has been either partially or entirely sequenced. PMID- 14558689 TI - Inhibition of plant-pathogenic fungi by the barley cystatin Hv-CPI (gene Icy) is not associated with its cysteine-proteinase inhibitory properties. AB - The recombinant barley cystatin Hv-CPI inhibited the growth of three phytopathogenic fungi (Botrytis cinerea, Colletotrichum graminicola, and Plectosphaerella cucumerina) and the saprotrophic fungus Trichoderma viride. Several mutants of barley cystatin were generated by polymerase chain reaction approaches and both their antifungal and their cysteine-proteinase inhibitory properties investigated. Point mutants R38-->G, Q63-->L, and Q63-->P diminished their capacity for inhibiting papain and cathepsin B, retaining their antifungal properties. However, mutant C68-->G was more active for papain and cathepsin B than the wild type. These results indicate that in addition to the consensus cystatin-reactive site, Q63-V64-V65-A66-G67, the A37-R38-F39-A40-V41 region, common to all cereal cystatins, and the C68 residue are important for barley cystatin activity. On the other hand, the K92-->P mutant is inactive as a fungicide, but still retains measurable inhibitory activity for papain and cathepsin B. Against B. cinerea, the antifungal effect of Hv-CPI and of its derived mutants does not always correlate with their activities as proteinase inhibitors, because the Q63-->P mutant is inactive as a cystatin, while still inhibiting fungal growth, and the K92-->P mutant shows the reciprocal effects. These data indicate that inhibition of plant-pathogenic fungi by barley cystatin is not associated with its cysteine-proteinase inhibitory activity. Moreover, these results are corroborated by the absence of inhibition of intra- and extramycelia-proteinase activities by barley cystatin and by other well-known inhibitors of cysteine-proteinase activity in the fungal zymograms of B. cinerea. PMID- 14558690 TI - Identical accumulation and immobilization of sulfated and nonsulfated Nod factors in host and nonhost root hair cell walls. AB - Nod factors are signaling molecules secreted by Rhizobium bacteria. These lipo chitooligosaccharides (LCOs) are required for symbiosis with legumes and can elicit specific responses at subnanomolar concentrations on a compatible host. How plants perceive LCOs is unclear. In this study, using fluorescent Nod factor analogs, we investigated whether sulfated and nonsulfated Nod factors were bound and perceived differently by Medicago truncatula and Vicia sativa root hairs. The bioactivity of three novel sulfated fluorescent LCOs was tested in a root hair deformation assay on M. truncatula, showing bioactivity down to 0.1 to 1 nM. Fluorescence microscopy of plasmolyzed M. truncatula root hairs shows that sulfated fluorescent Nod factors accumulate in the cell wall of root hairs, whereas they are absent from the plasma membrane when applied at 10 nM. When the fluorescent Nod factor distribution in medium surrounding a root was studied, a sharp decrease in fluorescence close to the root hairs was observed, visualizing the remarkable capacity of root hairs to absorb Nod factors from the medium. Fluorescence correlation microscopy was used to study in detail the mobilities of sulfated and nonsulfated fluorescent Nod factors which are biologically active on M. truncatula and V. sativa, respectively. Remarkably, no difference between sulfated and nonsulfated Nod factors was observed: both hardly diffuse and strongly accumulate in root hair cell walls of both M. truncatula and V. sativa. The implications for the mode of Nod factor perception are discussed. PMID- 14558691 TI - Differential defense reactions in leaf tissues of barley in response to infection by Rhynchosporium secalis and to treatment with a fungal avirulence gene product. AB - Expression of defense-associated genes was analyzed in leaf tissues of near isogenic resistant and susceptible barley cultivars upon infection by Rhynchosporium secalis. The genes encoding pathogenesis-related (PR) proteins PR 1, PR-5, and PR-9 are specifically expressed in the mesophyll of resistant plants, whereas a germin-like protein (OxOLP) is synthesized in the epidermis irrespective of the resistance genotype. Restriction-mediated differential display was employed to identify additional epidermis-specific genes. This resulted in the detection of another PR gene, PR-10, along with a lipoxygenase gene, LoxA, and a gene of unknown function, pI2-4, which are specifically induced in the epidermis of resistant plants. The gene encoding a putative protease inhibitor, SD10, is preferentially but not exclusively expressed in the epidermis. The fungal avirulence gene product NIP1 triggers the induction of the four PR genes only. At least two additional elicitors, therefore, must be postulated, one for the unspecific induction of OxOLP and one for the resistance specific induction of LoxA, pI2-4, and SD10. PR-10 expression can be assumed to be the consequence of NIP1 perception by epidermis cells. In contrast, gene expression in the mesophyll is likely to be triggered by an as yet unknown signal that appears to originate in the epidermis and that is strongly amplified in the mesophyll. PMID- 14558692 TI - The Medicago truncatula sucrose synthase gene MtSucS1 is activated both in the infected region of root nodules and in the cortex of roots colonized by arbuscular mycorrhizal fungi. AB - The MtSucS1 gene encodes a sucrose synthase (EC 2.4.1.13) in the model legume Medicago truncatula. To determine the expression pattern of this gene in different organs and in particular during root endosymbioses, we transformed M. truncatula with specific regions of MtSucS1 fused to the gusAint reporter gene. These fusions directed an induction to the vasculature of leaves, stems, and roots as well as to flowers, developing seeds, young pods, and germinating seedlings. In root nodules, strong promoter activity occurred in the infected cells of the nitrogen-fixing zone but was additionally observed in the meristematic region, the prefixing zone, and the inner cortex, including the vasculature. Concerning endomycorrhizal roots, the MtSucS1 promoter mediated strongest expression in cortical cells harboring arbuscules. Specifically in highly colonized root sections, GUS-staining was furthermore detected in the surrounding cortical cells, irrespective of a direct contact with fungal structures. In accordance with the presence of an orthologous PsSus1 gene, we observed a comparable regulation of MtSucS1 expression in the grain legume Pisum sativum in response to microbial symbionts. Unlike other members of the MtSucS gene family, the presence of rhizobial or Glomus microsymbionts significantly altered and enhanced MtSucS1 gene expression, leading us to propose that MtSucS1 is involved in generating sink-strength, not only in root nodules but also in mycorrhizal roots. PMID- 14558693 TI - Development of a lesion-mimic phenotype in a transgenic wheat line overexpressing genes for pathogenesis-related (PR) proteins is dependent on salicylic acid concentration. AB - In the course of coexpressing genes for pathogenesis-related (PR) proteins for a class IV chitinase and an acidic glucanase in transgenic wheat plants, we regenerated a wheat line that developed necrotic lesions containing dead cells in the T2 and subsequent generations. Lesion spots were detected at the booting stage (5- to 6-week-old plants) in lines homozygous for the transgene loci. In contrast, lesions were not observed in hemizygous transgenic lines or lines silenced for transgene expression, indicating a requirement for high levels of transgene expression for the development of the lesioned phenotype. Lesion development was associated with the accumulation of host-encoded PR proteins, e.g., chitinases, beta-1,3-glucanases, thaumatin-like protein, and production of reactive oxygen intermediates. F1 progeny of a cross between the lesion-plus transgenic line and wild-type nontransgenic plants produced progeny with a normal phenotype, while the F2 progenies segregated for the lesion phenotype. Salicylic acid (SA) levels in plants with the lesion-plus phenotype were found to be several times higher than controls and nearly double the levels in hemizygous transgenic plants that lack lesions. SA application activated lesion development in excised leaf pieces of these hemizygous transgenic plants. Similar activation of lesion development in control plants occurred only when high concentrations of SA were applied for prolonged periods. Transcripts for phenylalanine-ammonia lyase, which provides precursors of SA, were elevated in homozygous transgenic plants. Our data suggest that transgene-induced lesion-mimic phenotype correlates with enhanced SA biosynthesis. PMID- 14558694 TI - A mating-induced protein of Phytophthora infestans is a member of a family of elicitors with divergent structures and stage-specific patterns of expression. AB - Five members of an elicitor-like gene family from Phytophthora infestans were examined. The family was identified through the analysis of M81, a mating-induced gene. The predicted M81 product resembled a 42-kDa P. sojae glycoprotein known to elicit defense reactions in plants, including a host of P. infestans, potato. M81 was the most structurally and functionally divergent of the P. infestans genes compared with the P. sojae sequence. M81 lacked elicitor activity, had the lowest protein identity (47%), displayed mating-specific transcription, and had a novel C-terminal domain. The latter contained a 30-residue proline- and threonine-rich motif, which, remarkably, was tandemly repeated 24 to 36 times in different alleles. M81C, M81D, and M81E better resembled the P. sojae protein based on amino acid identity (63 to 75%) and conserved elicitor activity. M81C and M81D mRNA accumulated only during zoosporogenesis, while M81E expression was restricted to hyphae. M81B, an apparent pseudogene, was physically linked to M81. The protein products of each gene were predicted to be extracellular transglutaminases ranging in size from 436 to 1,607 amino acids. Genes with an elicitor, proline- and threonine-rich repeat, and both elicitor and repeat domains were widely distributed throughout Phytophthora infestans. These findings help explain the natural functions of elicitors in pathogen biology and plant microbe interactions. PMID- 14558695 TI - Cucumber mosaic virus infection transiently breaks dsRNA-induced transgenic immunity to Potato virus Y in tobacco. AB - Post-transcriptional gene silencing (PTGS), an intrinsic plant defense mechanism, can be efficiently triggered by double stranded (ds)RNA-producing transgenes and can provide high level virus resistance by specific targeting of cognate viral RNA. The discovery of virus-encoded suppressors of PTGS led to concerns about the stability of such resistance. Here, we show that Cucumber mosaic virus (CMV) is able to suppress dsRNA-induced PTGS and the associated Potato virus Y (PVY) immunity in tobacco. CMV suppression supported only a transient PVY accumulation and did not prevent recovery of the transgenic plants from PVY infection. CMV inoculation resulted in strongly increased transgene mRNA levels due to suppression of PTGS, but accumulation of PVY-specific small interfering (si)RNA was unaffected. However, PVY accumulation in previously immune plants resulted in increased PVY siRNA levels and transgene mRNA was no longer detected, despite the presence of CMV. Transgene mRNA returned to high levels once PVY was no longer detected in CMV-infected plants. Recovered and chronically CMV-infected tissues were immune to further PVY infection. PMID- 14558696 TI - Differences in the methyl ester distribution of homogalacturonans from near isogenic wheat lines resistant and susceptible to the wheat stem rust fungus. AB - Plants possess an efficient nonself surveillance system triggering induced disease resistance mechanisms upon molecular recognition of microbial invaders. Successful pathogens have evolved strategies to evade or counteract these mechanisms, e.g., by the generation of suppressors. Pectic fragments produced during host cell wall degradation can act as endogenous suppressors of the hypersensitive response in wheat leaves. We have isolated and characterized homogalacturonans from cell walls of two wheat cultivars susceptible to the stem rust fungus, Puccinia graminis f. sp. tritici, namely cvs. Prelude and Marquis, and from near-isogenic lines of both cultivars containing the Sr5-gene for hypersensitive rust resistance. Two independent approaches were used to compare their methyl esterification: i) immunochemistry using the monoclonal antibodies JIM5, JIM7, PAM1, and LM7 and ii) chromatography of oligogalacturonides representing stretches of contiguous nonmethyl-esterified GalA residues. The results clearly indicate a significant difference in the homogalacturonans from susceptible and resistant wheat lines. The difference can best be explained by assuming a nonrandom and more blockwise distribution of the methyl esters in the homogalacturonans of susceptible wheat cultivars as compared with a presumably more random distribution in the near-isogenic resistant lines. Possible consequences of this difference for the enzymatic generation of endogenous suppressors are discussed. PMID- 14558697 TI - Regulatory concerns regarding filters in pharmaceutical water systems. PMID- 14558698 TI - Development of guidance modules by the PDA to extend and/or eliminate periodic validation: a proposal to PDA Scientific Advisory Board. PMID- 14558699 TI - A method of increasing test range and accuracy of bioindicators: Geobacillus stearothermophilus spores. AB - Spores of Geobacillus stearothermophilus are very sensitive to changes in temperature. When validating sterilizing processes, the most common bioindicator (BI) is spores of Geobacillus stearothermophilus ATCC12980 and ATCC7953 with about 10(6) spores /BI and a D121-value of about 2 minutes in water. Because these spores of Geobacillus stearothermophilus do not survive at a F0-value above 12 minutes, it has not been possible to evaluate the agreement between the biological F-value (F(BIO)) and physical measurements (time and temperature) when the physical F0-value exceeds that limit. However, it has been proven that glycerin substantially increases the heat resistance of the spores, and it is possible to utilize that property when manufacturing BIs suitable to use in processes with longer sterilization time or high temperature (above 121 degrees C). By the method described, it is possible to make use of the sensitivity and durability of Geobacillus stearothermophilus' spores when glycerin has increased both test range and accuracy. Experience from years of development and validation work with the use of the highly sensitive glycerin-water-spore-suspension sensor (GWS-sensor) is reported. Validation of the steam sterilization process at high temperature has been possible with the use of GWS-sensors. It has also been shown that the spores in suspension keep their characteristics for a period of 19 months when stored cold (8 degrees C). PMID- 14558700 TI - Determination of in-process limits during parenteral solution manufacturing using Monte Carlo Simulation. AB - The purpose of this study is to utilize Monte Carlo Simulation methodology to determine the in-process limits for the parenteral solution manufacturing process. The Monte Carlo Simulation predicts the distribution of a dependable variable (such as drug concentration) in a naturally occurring process through random value generation considering the variability associated with the depended variable. The propagation of variation in drug concentration from batch to batch is cascading in nature during the following four formulation steps: 1) determination of drug raw material potency (or purity), 2) weighing of drug raw material, 3) measurement of batch volume, and 4) determination of drug concentration in the mix tank. The coefficients of variation for these four steps are denoted as CV1, CV2, CV3, and CV4, respectively. The Monte Carlo Simulation was performed for each of the above four cascading steps. The results of the simulation demonstrate that the in-process limits of the drug can be successfully determined using the Monte Carlo Simulation. Once the specification limits are determined, the Monte Carlo Simulation can be used to study the effect of each variability on the percent out of specification limits (OOL) for the in-process testing. Demonstrations were performed using the acceptance criterion of less than 5% of OOL batches, and the typical values of CV2 and CV3 being equal to 0.03% and 0.5%, respectively. The results show that for the in-process limits of +/- 1%, the values of CV1 and CV4 should not be greater than 0.1%. These assay requirements appear to be difficult to achieve for a given chemical analytical method. By comparison, for the In-process limits of +/- 4%, the requirements are much easier to achieve. The values of CV1 and CV4 should not be greater than 1.38%. In addition, the relationship between the percent OOL versus CV1 or CV4 is nonlinear per se. The number of OOL batches increases rapidly with increasing variability of CV1 or CV4. PMID- 14558701 TI - Accuracy of automated instruments used in the pharmaceutical industry for integrity testing sterilizing filters. AB - In the pharmaceutical industry, the integrity of sterile filters is critical to ensure sterility of filtered products. Filter integrity is frequently tested by measuring gas diffusion across water-contacting hydrophobic or hydrophilic membranes with the same automated test devices. Constant device accuracy over the whole range of possible operating conditions is an especially important requirement, as set by the GMP regulations for product critical devices. In this paper, we investigate the accuracy of gas diffusion rate and water intrusion rate estimates provided by a batch-operated and a refilling, continuous-flow commercial automated test device used both for diffusive flow tests and water intrusion tests. Tests were performed on custom-designed model filter systems and full-scale filters over a broad range of gas diffusive flow rates and upstream gas volumes. Neither tested device provided accurate measurements of gas diffusion rate when a small gas diffusion flow was measured out of a very large upstream volume. The batch-operated device provided measurements of gas diffusion rates (either gas diffusion or water intrusion rate) with an accuracy that strongly depends on the gas diffusion rate and on the gas volume upstream from the membrane. Gas diffusion rate measurements were particularly biased in diffusive flow tests of filters with less than 500 mL gas upstream volume. Gas diffusion rates were underestimated by as much as -14.5% in diffusive flow tests and -25% in water intrusion tests. The refilling, continuous flow device generally provided consistent and accurate gas diffusion rate and water intrusion rate measurements within less than 5% of the reference value, practically independent of the gas diffusion flow rate and upstream volume value. A serious bias was only noted in diffusion flow tests at very high upstream volumes and low gas diffusion rate. The results reported in this paper show the importance of qualifying the automated test devices used to assess sterile filter integrity. PMID- 14558702 TI - Extractables from filter components in model solvent streams for pharmaceutical filtration processes. AB - Drug manufacturers use filters widely in their manufacturing processes for bioburden reduction and sterility assurance. In implementing a pharmaceutical filtration, it is essential to determine and profile the filter extractables that could be introduced into the product. There is an ongoing industry effort to explore the analysis of extractables. Toward this end, the use of complementary analytical techniques was evaluated to aid in extractables identification by determining whether the analysis of filters in the form of components, rather than entire assemblies, facilitated the profiling of extractables. The study examines filter components of 10-inch, 0.22-microm Durapore cartridge filters using Fourier transform infra-red spectroscopy (FTIR) and nuclear magnetic resonance spectroscopy (NMR), as well as determines the non-volatile residues (NVR) and total organic carbon contents (TOC) of the extractables. The complementary techniques of high performance liquid chromatography (HPLC/UV) and liquid chromatography nuclear magnetic resonance spectroscopy (LC/NMR) were also used. The results of the tests indicated very low levels of extractables. PMID- 14558703 TI - An assessment of techniques for evaluating the physical stability of parenteral emulsions. AB - The physical stability of the parenteral emulsions is a key product quality issue. The purpose of this study is to develop, prepare and characterize model phospholipid emulsions and to critically evaluate various physical stability indicating methods. Oil-in-water (O/W) emulsions were prepared using 20% (w/w) medium chain triglycerides (MCT) or soybean oil in 2.21% (w/w) aqueous glycerin solutions emulsified with 0.1 to 1.8 % (w/w) lecithin. The reproducibility of emulsion preparation was determined by measuring the volume-based mean droplet diameter using photon correlation spectroscopy (PCS) and zeta potential using electrophoretic light scattering. Evaluation of stability-indicating methods was conducted by comparing the mean droplet growth rate of a thermally-stressed emulsion using PCS, a light obscuration particle counter (HIAC, equipped with a laser diode sensor) and a droplet image analyzer interfaced with transmission electron microscopy (TEM) using osmium tetraoxide fixation. Emulsions with identical compositions and preparation properties had reproducible mean droplet diameter and initial zeta potential values with RSD < 5.0%. Upon the application of thermal stress, the volume-based diameter increased linearly with time for all three sizing techniques (PCS, HIAC, and TEM). The droplet growth rates estimated using PCS and TEM were nearly identical. PCS is a sufficiently accurate technique for measuring emulsion stability and is less time-consuming than TEM. The HIAC technique only measured the size of droplets with diameters larger than about 1 micron, which was considerably greater than the mean droplet diameter as determined by PCS and microscopic image analysis (TEM). Moreover, the growth rate obtained using HIAC was much greater than the rates estimated by PCS and TEM; therefore the HIAC technique was not an accurate measure of the physical stability of the thermally stressed emulsions. PMID- 14558704 TI - Historical profile of Kurt Karl Stephan Semm, born March 23, 1927 in Munich, Germany, resident of Tucson, Arizona, USA since 1996. PMID- 14558705 TI - Desertification of the peritoneum by thin-film evaporation during laparoscopy. AB - OBJECTIVE: To assess the effects of gas flow during insufflation on peritoneal fluid and peritoneal tissue regarding transient thermal behavior and thin-film evaporation. The effects of laparoscopic gas on peritoneal cell desiccation and peritoneal fluid thin-film evaporation were analyzed. METHODS: Measurment of tissue and peritoneal fluid and analysis of gas flow dynamics during laparoscopy. RESULTS: High-velocity gas interface conditions during laparoscopic gas insufflation result in peritoneal surface temperature and decreases up to 20 degrees C/second due to rapid thin-film evaporation of the peritoneal fluid. Evaporation of the thin film of peritoneal fluid extends quickly to the peritoneal cell membrane, causing peritoneal cell desiccation, internal cytoplasmic stress, and disruption of the cell membrane, resulting in loss of peritoneal surface continuity and integrity. Changing the gas conditions to 35 degrees C and 95% humidity maintains normal peritoneal fluid thin-film characteristics, cellular integrity, and prevents evaporative losses. CONCLUSIONS: Cold, dry gas and the characteristics of the laparoscopic gas delivery apparatus cause local peritoneal damaging alterations by high-velocity gas flow with extremely dry gas, creating extreme arid surface conditions, rapid evaporative and hydrological changes, tissue desiccation, and peritoneal fluid alterations that contribute to the process of desertification and thin-film evaporation. Peritoneal desertification is preventable by preconditioning the gas to 35 degrees C and 95% humidity. PMID- 14558706 TI - Laparoscopic live donor nephrectomy: a step forward in kidney transplantation? AB - Open donor nephrectomy for live donor kidney transplantation is a safe procedure that has been used for more than 30 years with excellent results. Laparoscopic donor nephrectomy is a relatively new technique that has the potential of decreased postoperative pain, less incisional morbidity, and shorter recovery time. Furthermore, it has been reported that this potentially less traumatic approach increases the number of potential live donors. This review article focuses on the currently used laparoscopic techniques in live kidney donation as well as the controversy regarding its efficacy, safety, and future. PMID- 14558707 TI - New adhesion prevention concept in gynecological surgery. AB - Postsurgical adhesion formation is a common but significant problem. This is the first clinical evaluation of a new barrier material designed to reduce or eliminate postsurgical adhesions. SprayGel can be delivered laparoscopically or via laparotomy to form a strongly adherent hydrogel film. In this multicenter study, we evaluated the safety and effectiveness of SprayGel absorbable adhesion barrier system in patients undergoing open or laparoscopic myomectomy procedures. Here, we discuss the results of our evaluation conducted at the University of Kiel and Polyclinic of Bordeaux, and assess some of the features of this novel adjunct to prevent formation of postsurgical adhesions. PMID- 14558708 TI - Symptoms and antireflux medication use following laparoscopic Nissen fundoplication: outcome at 1 and 4 years. AB - BACKGROUND: We have reported 1-year outcomes and antacid medication use in 100 patients undergoing laparoscropic Nissen fundoplication. As a follow-up study, we queried these same patients to determine whether their outcomes endured 4 years after fundoplication. METHODS: One hundred patients undergoing laparoscopic Nissen fundoplication between 1992 and 1997 were asked, at 1 to 2 years and 4 to 6 years postoperatively, to grade their symptoms on a scale of 1 (mild) to 10 (severe). Patients were also queried as to the number/cost of antacid medications used before and after fundoplication. RESULTS: Significant improvements were noted in symptoms of heartburn, postprandial emesis, gas/bloating, and dysphagia after fundoplication. Significant decreases in antacid medication use (97% vs 19%) and monthly costs (dollar 168 +/- dollar 91 vs dollar 30 +/- 54) were seen following fundoplication. The number of patients on antacid medications and the monthly costs of these medications (37% and dollar 53 +/- dollar 87, respectively) increased significantly from early to late follow-up, but were still significantly lower than those before surgery. Overall, 87% and 900% of patients were pleased with their outcome at early and late follow-up, respectively, and 93% and 92% of patients stated they would consider undergoing fundoplication again if necessary (P = NS). CONCLUSION: Laparoscopic Nissen fundoplication results in a significant reduction in the symptoms of reflux and the use of antacid medications with a high degree of patient satisfaction. Although some patients return to antacid medications at late follow-up, they continue to have few symptoms and are pleased with their outcomes. PMID- 14558709 TI - Minimally invasive surgical treatment of esophageal achalasia. AB - BACKGROUND AND OBJECTIVES: A minimally invasive approach is considered the treatment of choice for esophageal achalasia. We report the evolution of our experience from thoracoscopic Heller myotomy (THM) to laparoscopic Heller myotomy (LHM). Our objective is to define the efficacy and safety of these 2 approaches. METHODS: Between March 1993 and December 2001, 36 patients underwent minimally invasive surgery for achalasia. Sixteen patients underwent THM without an antireflux procedure, and 20 patients underwent LHM with partial anterior fundoplication (n = 13) or closure of the angle of His (n = 7). RESULTS: Mean operative time and mean hospital stay were significantly shorter for LHM compared with that of THM (148.3 +/- 38.7 vs 222 +/- 46.1 min, respectively; P = 0.0001) and (2.06 +/- 0.65 days vs 5.06 +/- 0.85 days, respectively; P = 0.0001). Six of 16 patients (37.5%) in the THM group experienced persistent or recurrent dysphagia compared with 1 of 20 patients (5%) in the LHM group (P = 0.01). Heartburn developed in 5 patients (31.2%) after THM and in 1 patient (5%) after LHM (P = 0.06). Regurgitation developed in 4 patients (25%) after THM and in 2 patients (10%) after LHM (P = 0.2). Lower esophageal sphincter (LES) basal pressure decreased significantly from 30.1 +/- 5.07 to 15.3 +/- 2.1 after THM and from 31.8 +/- 6.2 to 10.4 +/- 1.7 after LHM (P = 0.0001). Mean esophageal diameter was significantly reduced after LHM compared with that after THM (from 53.9 +/- 5.9 mm to 27.2 +/- 3.3 mm vs 50.8 +/- 7.6 mm to 37.2 +/- 6.9 mm respectively: P = 0.0001). CONCLUSION: In our experience, LHM is associated with better short-term results and is superior to THM in relieving dysphagia. LHM with partial anterior fundoplication should be considered the treatment of choice for achalasia. PMID- 14558710 TI - Comparison of open and laparoscopic prosthetic repair of large ventral hernias. AB - BACKGROUND AND OBJECTIVES: Open ventral hernia repair is associated with significant morbidity and high recurrence rates. Recently, the laparoscopic approach has evolved as an attractive alternative. Our objective was to compare open with laparoscopic ventral hernia repairs. METHODS: Fifty laparoscopic and 22 open ventral hernia repairs were included in the study. All patients underwent a tension-free repair with retromuscular placement of the prosthesis. No significant difference between the 2 groups was noted regarding patient demographics and hernia characteristics except that the population in the open group was relatively older (59.4 vs 47.82, P < 0.003). RESULTS: We found no significant difference in the operative time between the 2 groups (laparoscopic 132.7 min vs open 152.7 min). Laparoscopic repair was associated with a significant reduction in the postoperative narcotic requirements (27 vs 58.95 mg i.v. morphine, P < 0.002) and the lengths of nothing by mouth (NPO) status (10 vs 55.3 hrs. P < 0.001), and hospital stay (1.88 vs 5.38 days, P < 0.001). The incidence of major complications (1 vs 4, P < 0.028), the hernia recurrence (1 vs 4, P < 0.028), and the time required for return to work (25.95 vs 47.8, P < 0.036) were significantly reduced in the laparoscopic group. CONCLUSIONS: Laparoscopic ventral hernioplasty offers significant advantages and should be considered for repair of primary and incisional ventral hernias. PMID- 14558711 TI - Methotrexate for the treatment of unruptured tubal pregnancy: a prospective nonrandomized study. AB - BACKGROUND AND OBJECTIVES: The aim of this study was to compare in a prospective nonrandomized study, the efficacy of 2 methods of administering methotrexate (MTX) in the treatment of ectopic pregnancy (EP): transvaginal injection under sonographic control or intramuscular injection (IM). METHODS: Patients with EP who met specific inclusion criteria for medical treatment were treated with MTX: 63 patients (group 1) were treated by IM and 47 patients (group 2) by transvaginal local injection. In group 1, 50 mg/m2 of MTX was injected intramuscularly; in group 2, transvaginal injection of 1 mg/kg of MTX was injected into the ectopic sac under sonographic control. When an additional dose of MTX was required, it was administrated IM at the dosage of 50 mg/m2 in both groups. RESULTS: The overall success rate, defined by a posttreatment normal hCG level (< 10 mUI/mL) was 71.4% in group 1 versus 91.5% in group 2 (P < 0.01); for patients with hCG levels < 2000 mUI/mL, 83% and 96%, respectively (not significant); for patients with hCG > or = 2000 mUI/mL, 37.5% and 86.4%, respectively (P < 0.01). CONCLUSION: In the medical treatment of EP, the efficacy of MTX is greater when administered by local transvaginal injection than by IM injection. We propose local treatment every time EP can be punctured, especially when hCG levels are > or = 2000 mUI/mL. PMID- 14558712 TI - Laparoscopic diagnosis and treatment in gynecologic emergencies. AB - OBJECTIVE: To present an analysis of our experience with 22 consecutive cases of acute abdominal gynecologic emergencies managed with a laparoscopic approach. METHODS: From March 1997 to October 1998, 22 patients with a diagnosis of acute abdominal gynecologic emergencies underwent laparoscopic intervention. A transvaginal ultrasound was performed on all patients preoperatively to supplement the diagnostic workup. Surgical time, complications, and length of hospital stay were evaluated, and the laparoscopic diagnosis was compared with the preoperative diagnosis. RESULTS: The laparoscopic diagnosis was different from the preoperative diagnosis in 31.8% of patients. Of the 22 patients, laparoscopic therapeutic procedures were performed in 18 (81.8%), all satisfactorily, and with no need for conversion to open surgery. No morbidity or mortality occurred. CONCLUSION: Laparoscopy is a safe and effective method for diagnosing and treating gynecologic emergencies. PMID- 14558714 TI - Liver cyst with biliary communication successfully treated with laparoscopic deroofing: a case report. AB - A 71-year-old Japanese woman complained of right upper abdominal fullness and pain. Computed tomography revealed a huge cyst in the right lobe of the liver, measuring 16 cm in diameter. She underwent laparoscopic deroofing of the liver cyst. On operation, needle aspiration of the cyst yielded clear serous fluid without any bile contamination. However, after the cyst was deroofed with laparoscopic coagulating shears, bile leakage was recognized from a tiny orifice in the cyst cavity. A catheter was inserted via the orifice for cholangiography, which demonstrated a communication with the biliary tract. The orifice was easily closed with a laparoscopic suturing device. Operation time was 5 hours and 30 minutes, and blood loss was 300 grams. Pathological examination of the liver cyst was consistent with a simple cyst. The postoperative course was uneventful, and the patient has had no recurrence to date at 13 months. Laparoscopic deroofing is a recommended treatment for a liver cyst even in the presence of cystobiliary communication. PMID- 14558713 TI - Surgeon-led initiatives cut costs and enhance the quality of endoscopic and laparoscopic procedures. AB - BACKGROUND: Public perception depicts surgical cost control and quality of care as polar opposites. We describe a program led by practicing surgeons that demonstrates that quality can be maintained, and often improved, while substantial cost reductions are realized. METHODS: A set of evidence-based protocols was developed, revised, and followed for 42 procedures in general, otolaryngologic, urologic, and orthopedic surgery. Each protocol consists of surgeon-initiated guidelines on operative indications, preoperative testing, preadmission planning, length of stay, resource utilization, convalescence, and pharmacy services. Information was collected for 24 months from July 1998 to July 2000 by 62 surgeons in Kentucky and Indiana. Data were obtained for 4302 cases, among them colonoscopy (1145), esophagogastroduodenoscopy (714), laparoscopic cholecystectomy (418), endoscopic retrograde cholangiopancreatography (235), and laparoscopic fundoplication (87). RESULTS: Specific cost reductions occurred in laparoscopic cholecystectomy by limiting the administration of perioperative antibiotics. Sixty-seven percent of patients did not receive antibiotics. Outpatient cholecystectomy was the norm (60%), due primarily to preadmission planning through discussion with the patients and their family. Interestingly, when surgeons were educated on the costs of certain instruments and medications, their practices changed. The avoidance of a particular postoperative antiemetic, which was more than tenfold more expensive than other choices, was rapidly adopted by all surgeons when the costs were discovered. One participating teaching hospital used its own financial data and predicted that if all surgeons at their facility followed the protocols and had similar results, a savings of dollar 1.1 million per quarter would be obtained. PMID- 14558715 TI - Laparoscopy for treating a small bowel obstruction due to a Meckel's diverticulum. AB - A Meckel's diverticulum is one cause of small bowel obstruction in the virgin abdomen. A 17-year-old female presented with a 24-hour history of lower abdominal pain and emesis. Radiological imaging studies revealed a high-grade partial small bowel obstruction. A diagnostic laparoscopy was performed revealing a bowel obstruction secondary to a Meckel's diverticulum. The diverticulum was resected using an endovascular GIA stapler. The patient was discharged on postoperative day 3, tolerating a regular diet. Laparoscopy is a useful diagnostic and therapeutic tool for a patient with a small bowel obstruction with an unclear etiology. PMID- 14558716 TI - Laparoscopic treatment of ovarian vein syndrome. AB - BACKGROUND: Ovarian vein syndrome is a rare cause of teral obstruction. In this report, we describe an unusual presentation of the syndrome successfully treated with laparoscopic techniques. METHODS: The patient presented with a 12-month history of right flank pain and a right abdominal mass. The preoperative evaluation revealed renal malrotation, hydronephrosis, decreased renal function, and presumed ureteropelvic junction obstruction. RESULTS: By using a transperitoneal laparoscopic approach, an enlarged ovarian vein was identified as the cause of the ureteral obstruction. The ovarian vein was divided with a laparoscopic stapler. The patient's postoperative course was unremarkable, and she was discharged from the hospital on the second postoperative day. At 3-months follow-up, the patient was completely asymptomatic without evidence of obstruction. DISCUSSION: Ovarian vein syndrome remains a rare diagnosis of exclusion. A careful preoperative evaluation is required to exclude other causes of ureteral obstruction. By using a laparoscopic approach, the ureter and obstructing vessel were readily identified to effectively treat the patient. With the minimally invasive approach, postoperative recovery and patient quality of life were improved. PMID- 14558717 TI - Laparoscopic removal of an artery forceps. AB - Numerous foreign bodies, such as surgical gauze, pads and instruments, and other items, have been left behind in the abdominal cavity during open surgeries. These have been traditionally removed at redo open surgeries. Here we describe a case of an artery forceps left behind at a previous surgery (open cholecystectomy and appendicectomy) performed 5 years earlier that was removed by laparoscopy. PMID- 14558718 TI - Laparoscopic retroperitoneal lymph node dissection in the extremely obese patient: technical insight into access and port placement. AB - PURPOSE: We report on laparoscopic retroperitoneal lymph node dissection (RPLND) in a morbidly obese patient to discuss the associated technical steps for satisfactory completion of staging lymphadenectomy. METHODS: A laparoscopic RPLND was performed using a modified template on the left side. Initially, 4 ports were placed with the patient in the supine position. Three were placed 3 cm to the left of midline and one in the anterior axillary line, at the level of the umbilicus. During the operation, successful bowel retraction necessitated placement of 2 additional ports in the anterior axillary line (just above the pelvis and off the tip of the 12th rib). Using these 6 trocar sites, the dissection was completed, and 44 lymph nodes were obtained. RESULTS: Laparoscopic retroperitoneal lymph node dissection was accomplished in an extremely obese patient with acceptable morbidity by using prudent modification of standard techniques. CONCLUSION: If access and port placement limitations are overcome, the benefits of laparoscopy in the obese are clear. This report serves as a signpost that laparoscopic retroperitoneal lymph node dissection for testes cancer can also be accomplished using modification of standard techniques. PMID- 14558719 TI - Postoperative chylous ascites: a rare complication of laparoscopic Nissen fundoplication. AB - The accumulation of chylous fluid in the abdominal cavity is an infrequent, yet alarming, complication in abdominal surgery. Laparoscopic fundoplication has assumed a central role in the surgical treatment of gastroesophageal reflux disease and is significantly altering the balance of therapy toward more common and earlier surgical intervention. We report the case of a 67-year-old woman with gastroesophageal reflux disease and intense esophagitis who underwent a laparoscopic Nissen fundoplication in February 2000. The procedure was performed without apparent complications. Twenty days later, the patient complained of abdominal pain and distension. Ultrasonography showed ascites, whereas endoscopic and radiological exploration of the fundoplication demonstrated no abnormalities. A paracentesis was performed, which showed a milky fluid with high concentrations of triglycerides (1024 ng/dL) and cholesterol (241 ng/dL). The patient was treated successfully with total parenteral nutrition for 3 weeks, followed by a low-fat diet. To our knowledge, this is the third reported case of chylous ascites after a Nissen fundoplication and the second case after laparoscopic fundoplication. The development of chylous ascites seems to be related to the injury of lymphatic vessels, including the thoracic duct, during the retroesophageal window dissection. The careful dissection and judicious use of diathermy is proposed to prevent this rare complication. PMID- 14558720 TI - Laparoscopic bilateral adrenalectomy with the use of a single midline hand-assist port. AB - Indications for bilateral adrenalectomy are rarely present in patients with Cushing's syndrome. The laparoscopic approach to adrenalectomy provides a postoperative course that compares favorably with that of open adrenalectomy, and the hand-assisted technique may provide an additional alternative to the open approach in performing bilateral adrenalectomy. PMID- 14558721 TI - Jaundice due to extrabiliary gallstones. AB - OBJECTIVE: Cholecystectomy is one of the most common general surgical procedures performed today. The laparoscopic approach is beneficial to patients in terms of length of stay, postoperative pain, return to work, and cosmesis. Some drawbacks are associated with the minimal access form of cholecystectomy, including an increased incidence of common bile duct injuries. In addition, when the gallbladder is inadvertently perforated during laparoscopic cholecystectomy, retrieval of dropped gallstones may be difficult. We present a case in which gallstones spilled during cholecystectomy, causing near circumferential, extraluminal common hepatic duct compression, and clinical jaundice 1 year later. METHODS: The patient experienced jaundice and pruritus 12 months after laparoscopic cholecystectomy. A computed tomographic scan was interpreted as cholelithiasis, but otherwise was normal (despite a previous cholecystectomy). Endoscopic retrograde cholangiopancreatography was performed and a stent placed across a stenotic common hepatic duct. RESULTS: The results of brush biopsies were negative. The stent rapidly occluded and surgical intervention was undertaken. At exploratory laparotomy, an abscess cavity containing multiple gallstones was encountered. This abscess had encircled the common hepatic duct, causing compression and fibrosis. The stones were extracted and a hepaticojejunostomy was tailored. The patient's bilirubin level slowly decreased and she recovered without complication. CONCLUSIONS: Gallstones lost within the peritoneal cavity usually have no adverse sequela. Recently, however, numerous reports have surfaced describing untoward events. This case is certainly one to be included on the list. A surgeon should make every attempt to retrieve spilled gallstones due to the potential later complications described herein. PMID- 14558722 TI - Scissor-knot-pusher: an instrument for simplified laparoscopic extracorporeal knotting. AB - This paper describes the "scissor-knot-pusher," an instrument that greatly facilitates the execution of knot tying during laparoscopic operations. The instrument acts in essence as an extension of the surgeon's hand and, given its rigid structure, allows the surgeon full control of the process of knot tying. Additionally, after the knot has been tightened, it is possible to cut the suture without using a different instrument. As a result, this technical device simplifies knot tying and may help to reduce the frustration and the time often associated with intracorporeal suturing during laparoscopic surgery. PMID- 14558723 TI - Solitary hepatic gastrinoma treated with laparoscopic radiofrequency ablation. AB - BACKGROUND: This is a case of a solitary hepatic gastrinoma in a 65-year-old male. The patient was diagnosed with Zollinger-Ellison syndrome in 1991. He had negative radiologic and surgical explorations at that time. He was maintained on proton-pump inhibitors for the next 10 years without symptoms. METHODS: A computed tomographic (CT) scan done in April 2001 demonstrated a 5-cm right hepatic lesion. Radionucleotide scanning with octreotide demonstrated intense activity in the same area in the right hepatic lobe. His serum gastrin was 317 pg/mL. He underwent laparoscopic radiofrequency ablation of the lesion. RESULTS: Treatment resulted in a 6-cm ablative area giving a 1-cm margin on the tumor. One and 3-month follow-up CT scans demonstrated adequate ablation of the tumor. An octreotide scan done 3 months postoperatively did not reveal any areas of abnormal uptake. CONCLUSION: We report success with laparoscopic radiofrequency ablation as an alternative to major hepatic resection in patients with a solitary hepatic gastrinoma. PMID- 14558724 TI - Prospects for gene therapy in hearing loss. AB - Deafness is the most common form of sensory impairment in humans. Depending on the age of onset, hearing impairment can affect oral language acquisition, cognitive development and psychosocial development. Here, we cover the latest advances in gene therapy for alleviating or preventing hearing loss. This review is not meant to be comprehensive, but to highlight some of the most recent developments in the field. Several recent reviews have described potential therapeutic approaches. PMID- 14558725 TI - The differential vulnerability of the inner ear end-organs to several external factors. AB - In a number of recently conducted animal studies, the effect of various external factors such as ototoxic substances, different types of noise and systemic disease on the different end-organs of the inner ear has been investigated. These studies are distinguished by the use of short latency vestibular evoked potentials (VsEPs) (to both linear and angular acceleration), an objective method for directly assessing the function of the different vestibular end-organs. In addition, the well known auditory brainstem response (ABR) was used to assess cochlear function. The studies are reviewed and it appears that the general pattern of effect is as follows: ABR (cochlea) is the most sensitive to the various external factors; angular VsEPs (semicircular canals) the least sensitive; linear VsEPs (otolith organs) intermediate between them. PMID- 14558727 TI - Preventing noise induced otoacoustic emission loss by increasing magnesium (Mg2+) intake in guinea-pigs. AB - This study aimed to explore the effects of increased Mg2+ intake on the activity of the outer hair cells as manifested in Click and Distortion Evoked Otoacoustic Emissions (CEOAEs; DPOAEs) in 25 animals. Thirteen animals were fed with high Mg2+ intake (39 mmol Mg2+/l in drinking water) and 12 without the Mg2+ additive. The OAE amplitudes and frequency ranges as well as the DPOAE thresholds were affected significantly less by noise exposure in the animals fed Mg2+-enriched water. Following the exposure, the auditory recovery was faster in the high than the low Mg2+ animals (controls). In addition, a relationship was found between the Mg2+ level and the emission loss. The post-exposure measures may result from the effect of Mg2+ on cochlear metabolic processes and vascular microcirculation. The results demonstrate that pre-existing low Mg2+ levels will exacerbate noise induced hearing loss (NIHL), and increased Mg2+ intake provides a significant biological cochlear protective effect. PMID- 14558726 TI - Use of ABR threshold and OAEs in detection of noise induced hearing loss. AB - OBJECTIVE: To determine which measure is the most sensitive to noise induced hearing loss (NIHL): auditory nerve brainstem response (ABR), distortion product otoacoustic emission (DPOAE) or transient evoked otoacoustic emission (TEOAE), and how to assess possible changes in these responses. SUBJECTS & METHODS: Four groups of rats were exposed to various durations of 113 dB SPL broadband noise: 5 or 10 minutes (temporary changes in cochlear function), and 3 or 4 hours (permanent changes). Group means and data from individual animals were compared before and after exposure. RESULTS: Mean group DPOAE amplitude reduction showed no clear advantage over mean ABR threshold elevation in detection of temporary and permanent NIHL. Data from individual rats, however, indicated a clinical advantage for DPOAEs in detecting slight temporary, but not permanent, changes. TEOAEs were more sensitive in detecting changes in individual rats than as a group measure. CONCLUSIONS: TEOAE and DPOAE monitoring may improve detection of NIHL, though it should be used in conjunction with audiometric threshold monitoring. PMID- 14558728 TI - The effect of training on frequency discrimination: generalization to untrained frequencies and to the untrained ear. AB - While there is growing evidence that frequency discrimination improves with practice, there are, however, limited and inclusive reports regarding the generalization of learning to untrained conditions. The goals of the present study were therefore (1) to measure the effect of multi-session training on difference limen frequency (DLF) thresholds and evaluate the relative contribution of procedural and stimulus learning by comparison to a control (untrained) group; (2) to evaluate the extent of generalization of the trained frequency to two untrained frequencies known to be temporally coded; and (3) to estimate the generalization of the trained frequency to the untrained ear, for both trained and untrained frequencies. Two groups of subjects were included: a trained group (n = 5) and a control group (n = 5). For the trained group, DLF thresholds for 1 kHz (trained frequency) and 1.1 and 2.0 kHz (untrained frequencies) were obtained using a two-interval, two-alternative forced choice paradigm before and after an eight-session training. The control group was tested using the same stimuli only twice, with a 3-week interval between testing. Results showed that (1) multi-session training improves frequency discrimination in normal hearing young adults; (2) adding a control group to the study allowed estimation of the effect of limited exposure to the stimuli and task in naive listeners and evaluation of the magnitude of procedural learning; (3) learning was generalized across frequencies that are coded by similar mechanisms; (4) generalization of learning occurred in the untrained ear for trained and untrained frequencies. These results have important clinical and theoretical implications regarding the processes underlying perceptual learning and the effectiveness of auditory habilitation strategies. PMID- 14558729 TI - Effect of second-formant transitions on the perception of Hebrew voiced stop consonants. AB - Studies in English, Dutch, Danish and French show that of the possible acoustic cues that listeners use for the perception of place of articulation, the transition of the second formant (F2) appears to be a very important cue. Although the Hebrew language shares some similarities with the above languages, one cannot assume that it either has similar acoustic-articulatory patterns or uses the same cues for perception. The general goal of the present study was, therefore, to investigate the effect of the starting frequency of F2 transition on the perception of place of articulation of Hebrew voiced plosives in initial position. Sixteen Hebrew-speaking young normal-hearing adults served as subjects. Stimuli were generated by re-synthesizing a naturally spoken /ba/ syllable into 17 test syllables by varying only the starting frequency of F2. Listeners heard each stimulus six times (total of 102) at random and were required to label the stimuli as /ba/, /da/ or /ga/. Results showed that varying only F2 transitions caused a perceptual change of place of articulation for all listeners. There was, however, large inter-subject variability in the perceived category: 75% of the subjects identified /ba/ and /da/, half of them also identified /ga/, and 25% of the subjects were able to identify /ba/ and /ga/ only. These data suggest that while F2 transitions are important for perceiving place of articulation of Hebrew voiced stops, they cannot predict the perceived category. It also supports the notion that normal-hearing listeners differ in the relative importance they assign to the cues for the perception of place of articulation. PMID- 14558730 TI - Effect of voice onset time (VOT), stop burst and vowel on the perception of voicing in Hebrew stops: preliminary results. AB - Very few studies investigated systematically the acoustic cues for the perception of voicing stops in Hebrew. Voicing is characterized by several parameters of which the voice onset time (VOT) was found to be the primary cue for its perception. There are, however, other known acoustic cues to voicing such as transition to the first formant (F1) and the initial burst. The purpose of the present study was to measure the relative weighting of these various acoustic cues in the perception of Hebrew voicing, using the conflicting cues paradigm. Thirteen adults with normal hearing participated in this study. Stimuli consisted of one pair of meaningful words that differ in the voicing of the initial stop. Four different continua were constructed from the pair of natural stimuli. The first two consisted of the voiced burst combined with the vowel that was truncated from the consonant-vowel combination (where the consonant was voiced or voiceless). The remaining two continua consisted of the voiceless burst combined with the same truncated vowels. For each stimulus, a VOT continuum was created varying from -40 to +40 ms in 10 ms segments. Subjects were tested using a two alternative forced choice labeling procedure. The percent of responses to each stimulus of each VOT continuum (/b-p/) was calculated for each individual and combination. The results show that each acoustic cue contributed to the perception of initial voicing in Hebrew: (1) When the stimulus was constructed from the voiced cues, positive VOT values were needed for the voice/voiceless distinction; (2) when the stimulus was constructed from the voiceless cues, negative VOT values were needed for the voicing distinction; and (3) when the stimulus was constructed from voiced and voiceless cues, intermediate VOT values were needed for the voicing distinction. These results provide initial information regarding the relative effect of the acoustic cues in the perception of Hebrew stop voicing. PMID- 14558731 TI - Semantic processing of unattended words and pseudowords in first and second language: an ERP study. AB - We recorded event related brain potentials to assess stages of linguistic processing of first (L1) and second (L2) language and of pseudowords when subjects were engaged in a different task and did not attend to the words. Young adults (n = 15) were presented with pairs of auditory stimuli consisting of words and pseudowords in L1 and L2 with different voice onset times (VOT), which served as distracters in a short-term memory task. ERPs were recorded from 11 scalp electrodes. The ERP peak amplitudes and latencies were subjected to analysis of variance for the effects of language, meaning and scalp location as well as priming of the second word in the pair by the preceding word. Behavioral results showed that attention was drawn to the primary task and away from the words; yet significant, including semantic, processing was evident in the ERPs to the words, with significant effects of language, meaning and priming. Even with barely any awareness of the stimuli, the brain processes words including distinguishing between L1 and L2 and relating to the stimuli's context. PMID- 14558732 TI - Tuberculosis in Pakistan: are we losing the battle? PMID- 14558733 TI - Differences in clinical presentation of pulmonary tuberculosis in association with age. AB - OBJECTIVE: To study the differences in presentation of pulmonary tuberculosis in young adult and elderly patients. DESIGN: A prospective study was conducted between December 1999 to May 2000, which included all the patients presenting with pulmonary tuberculosis at the Department of Thoracic Medicine, Jinnah Postgraduate Medical Centre (JPMC), Karachi. PATIENTS AND METHODS: There were 67 young adult (mean age 30.63 yrs) and 36 elderly patients (mean age 65.92 yrs) with pulmonary tuberculosis. The difference in presentation of two groups were analyzed for statistical difference. Chi-square test was used for testing difference of percentage. The students t-test was used for testing difference of mean. The P<0.05 level of significance was adopted. RESULTS: The elderly patients were more likely to have dyspnoea (73% vs 23.9% P<0.001) and non-specific symptoms (62.2% vs 17.9% P<0.001) but less haemoptysis (21.6% vs 46.3% P<0.01). The chest radiograph in elderly patients more commonly had extensive bilateral infiltration (32.4% vs 14.9% P<0.03) and lower zone infiltration (37.8% vs 3% P<0.001). CONCLUSION: The result of our study suggests that elderly patients with pulmonary tuberculosis were more likely to present with dyspnoea, non-specific symptoms and atypical radiographic appearance. PMID- 14558734 TI - Is Ministry of Health fully prepared to implement an effective DOTS program in Pakistan? An operations research on TB control program in the public health sector in Sindh. AB - PURPOSE OF THE STUDY: Pakistan is among the high-burden countries for tuberculosis. One of the fundamental problems in TB control is a high defaulter rate among the registered TB cases in the public sector. In 1999, a cross sectional study was designed to identify the determinants of low compliance for the TB treatment in two rural districts in Sindh. METHODS: Before the actual data collection, a pilot testing was planned in a secondary level care hospital. Fourteen defaulters for TB treatment were identified but none could be contacted due to incomplete addresses. Other alternatives were explored with the health facility team to reach them including a field-based search through Lady Health Workers of the National Health Program but all endeavors went into vain. The pilot testing propelled us to postpone the cross-sectional study but we continued scrutinizing the follow up problem for TB patients in other health facilities. Not surprisingly, more or less a similar picture was found in those health facilities. PRINCIPAL CONCLUSIONS: The study concludes that the public health care system in Pakistan lacks even the basic requirements for an effective TB control program, that is, a viable information system and the functional integration of program with rest of the health care delivery system. A DOTS strategy to control TB was initiated in the public sector in Pakistan just one year prior to this study. The Ministry of Health requires re-visiting the program to ensure that the lacunae identified in this study are being taken care of in the current DOTS strategy. PMID- 14558735 TI - Frequency and antibiotic susceptibility pattern of mycobacterial isolates from extra-pulmonary tuberculosis cases. AB - OBJECTIVE: To determine the frequency and antimicrobial susceptibility pattern of extra-pulmonary tuberculosis in Rawalpindi. SETTING: Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi. METHODS: Between September 2000 and August 2002, 899 pulmonary and 460 extra-pulmonary specimens from suspected cases of tuberculosis were examined. The radiometric BACTEC 460 TB system was used for culture and antimicrobial susceptibility testing. RESULTS: Mycobacteria were isolated from 291 pulmonary specimens and 98 extra-pulmonary specimens. The frequency of extra-pulmonary tuberculosis was 25.2%. The commonest source of isolation was pus (44.9%, frequency 11.3%), followed by lymph nodes (13.3%, frequency 3.3%) and pleural fluid (13.3%, frequency 3.3%). We tested the anti microbial susceptibility of the isolates to the four first line anti-tuberculous drugs, rifampicin, isoniazid, streptomycin and ethambutol. Of the extra-pulmonary isolates 13.3% were resistant to a single drug, 21.4% were multi-drug resistant and 9.2% were resistant to all the four drugs. CONCLUSION: Increased awareness of the magnitude of the problem posed by extra-pulmonary tuberculosis is required so that appropriate control measures can be adopted. PMID- 14558736 TI - Addition of steroids in medicated dentrifices marketed in Pakistan: a possible serious health hazard. AB - OBJECTIVE: To determine whether some dentrifices contain steroids. MATERIALS AND METHODS: Samples of 20 brands claiming to have any medicinal benefit and readily available in the open market were submitted for testing for the presence of steroids to the Pakistan Council of Scientific and Industrial Research Laboratories, Karachi. RESULTS: Eight samples tested positive on the first test and seven were positive after a second, confirmatory test. CONCLUSION: All these brands are freely available over the counter and their labels do not list steroid as an ingredient. Exposing consumers unknowingly to steroid may be regarded as a serious health hazard. Our state regulatory bodies and media need to play an active role in the prevention and monitoring of such possibly unethical marketing. PMID- 14558737 TI - Re-emergence of Vibrio cholerae O139 in Pakistan: report from a tertiary care hospital. AB - OBJECTIVE: This study reports re-emergence of Vibrio cholerae O139 in Pakistan in 2000-2001 from a tertiary care hospital in Karachi, Pakistan. METHODS: This descriptive study was conducted from 2000-2001. Stool samples were taken from inpatients or those referred to the laboratory from other hospitals, clinics and general practitioners. Samples were processed and Vibrio cholerae isolates were identified according to standard protocols. Tellurite Taurocholate Gelatin agar was used as a selective medium for Vibrio cholerae. Serogroups were identified by slide agglutination with polyvalent antisera. Antimicrobial sensitivities were performed by Kirby Bauer technique. Data was entered and analyzed using SPSS, p values were calculated using t test and two independent samples test. RESULTS: During the study period, 144 samples were found to be infected with Vibrio cholerae O139 in comparison with 545 Vibrio cholerae O1. Infection with O139 was characteristically observed in the older population (mean age = 40 years) in contrast with Vibrio cholerae O1 strains (mean age = 23 years) (p. value = <0.001). Sensitivity pattern of 2000-2001 Vibrio cholerae isolates was markedly different to that of 1993-1994. The earlier isolates were resistant to Cotrimoxazole (99%) and Chloramphenicol (35%) whereas the recent isolates are almost 100% sensitive. CONCLUSION: In conclusion this re-emergent strain seen 6 years after previous episode infected large number of people especially older population suggesting that prior infection with O1 does not provide immunity against O139 and therefore Vibrio cholerae O139 has a potential to cause a major epidemic in an immunologically naive population. PMID- 14558739 TI - Eclampsia, a major cause of maternal and perinatal mortality: a prospective analysis at a tertiary care hospital of Peshawar. AB - OBJECTIVE: To observe pregnancy outcome in eclamptics and to explore the avoidable factors contributing to the adverse outcome. SETTING: Department of Obstetrics and Gynaecology, Hayatabad Medical Complex, Peshawar. METHOD: This prospective cross-sectional observational study (from 1st Jan 2001 to 31st Mar 2002) included all the patients admitted to the unit with eclampsia. A Sociodemographic and clinical data was collected along with the results of investigations to categorize the complications and data was analyzed using various statistical tests. MAIN OUTCOME MEASURES: Maternal and Perinatal morbidity and mortality. RESULTS: During the study period 71 patients developed eclampsia (frequency: 1.2%). Majority were unbooked (86%), primigravida (69%), < or = 25 years of age (63%), referred from other health facilities (66.2%) and had some delay in seeking medical help (60%). Thirty five percent of patients developed major complications and 16.9% of them died (48% of overall maternal mortality). Mortality was frequent in Afghani women (OR 7.71 p value 0.002) and in women who sought medical help more than 6 hrs after developing seizures (OR 14.6 P value 0.0004). Perinatal mortality was 41.6% having prematurity the main risk factor (OR 13.33 p value 0.0000068). CONCLUSION: To decrease the adverse outcome associated with eclampsia a community-based approach is needed to improve community health education, socioeconomic status and prenatal care. Delivery of proper health care system and emergency obstetrical care facilities are vital for prevention, early detection, proper management and hence to save the mothers and their babies from such a dreadful disease. PMID- 14558738 TI - Cefaclor AF vs Clarithromycin in acute exacerbation of chronic bronchitis (B3M-PK AJBG). AB - OBJECTIVE: To compare the efficacy and safety of Cefaclor AF vs Clarithromycin in the treatment of acute exacerbation of chronic bronchitis in adult subjects. PATIENTS AND METHODS: This study was conducted on 300 patients suffering from acute exacerbation of chronic bronchitis, who attended the out patient clinics of ten different hospitals throughout Pakistan. Pneumonia, bronchiectasis and tuberculosis were excluded with the help of chest radiography and sputum smear examination. Pretherapy sputum culture and sensitivity (c/s) were tested and patients were randomized and supplied with either tablet Cefaclor 375 mg or tablet Clarithromycin 250 mg to be taken twice daily. Patients were evaluated at day 0 and then at day 3-5 and day 10-11. Post therapy sputum c/s was done on day 10-11. A fourth and final visit was planned on day 20-24 which was optional. At each visit, the severity of disease and the signs and symptoms were recorded on the clinical report forms according to the preset standards. RESULTS: Of 136 patients in the Cefaclor group and 142 patients in the Clarithromycin group, cure was achieved in 44 vs 35 subjects, improvement in 78 vs 91 subjects and failure in 16 vs 18 subjects among Cefaclor vs Clarithromycin groups respectively. The overall clinical efficacy (cure and improvement combined ) was 88.4% in the Cefaclor group and 87.5% in the Clarithromycin group. Nine patients in the Cefaclor group and patients in the Clarithromycin group had one adverse event whereas twelve patients in each group had two or more adverse events. CONCLUSIONS: The above results show that both Cefaclor AF and Clarithromycin are equally effective and safe in the treatment of acute exacerbation of chronic bronchitis in adult patients. PMID- 14558740 TI - Institution based tumor registry from Punjab: five year data based analysis. AB - INTRODUCTION: There are no population based cancer registries in Pakistan except for Karachi Cancer Registry (KCR). No data from Punjab is available in the last decade. We decided to conduct a data base analysis to see the frequencies of different cancers in Punjab. PATIENTS AND METHODS: A retrospective analysis of 5100 patients presenting to Jinnah Hospital between January 1997 to December 2001 with histological diagnosis of cancer was performed. A tumor registry performa on all patients was filled which included demographic data as age, sex, duration of symptoms and disease stage at presentation. All cancers were coded according to international classification of Disease-oncology (ICD-10) classification. RESULTS: Male cancers accounted for 47.7% and female cancers 52.2% of the total cases. In males the three most frequent cancers were leukemias, non-Hodgkins lymphoma and lung cancer. In females breast cancer was the most common cancer accounting for 38.5% female followed by ovarian cancer 13.6%. CONCLUSIONS: Our institution based cancer registry indicates hematological malignancies are the most common cancers in males in Punjab followed by lung and colorectal cancer. In females breast cancer is the commonest cancer followed by ovarian cancer. Gall bladder cancer is relatively more frequent in women while cervical cancer is less commonly observed in our patient population. Despite biases until population based incidences are available, studies like ours may prove beneficial for future health planning and research. PMID- 14558741 TI - Diagnosis of pulmonary embolism with helical C.T. scan. AB - OBJECTIVE: To determine the role of helical C.T. scan in the diagnosis of pulmonary embolism. SETTING: Radiology department, Aga Khan University Hospital. One year prospective study May 2001 to May 2002. METHODS: Sixty patients suspected of having pulmonary embolism were included in this study. Helical C.T. scan chest was performed from aortic arch to lung bases with intravenous contrast medium. Images were acquired in mediastinal and lung windows. Image interpretation was performed on the console and hard copies for main branch and segmental thrombus. RESULTS: Fifteen patients were diagnosed of having pulmonary embolism on C.T. scan. Clinical correlation and follow up with pulmonary angiography and Doppler ultrasound of leg was carried out. Pulmonary angiography was performed in one patient with embolectomy done after confirmation of diagnosis. 8 patients had associated DVT confirmed on Doppler sonography. Sensitivity of helical C.T. is 93.75%, specificity 100% and positive predictive value 95.65%. CONCLUSION: Helical CT scan is a quick, easy and accurate imaging modality for diagnosing pulmonary embolism. PMID- 14558742 TI - What is wrong with medical audit? PMID- 14558743 TI - Post dural puncture headache. PMID- 14558744 TI - Treatment failure of tuberculosis due to concomitant pathology. PMID- 14558745 TI - A young woman with cough and abnormal chest radiograph. PMID- 14558746 TI - Isolated congenital bilateral absence of vas deferens (CBAVD): a rare anomaly and diagnostic dilemma. PMID- 14558747 TI - Localized melioidosis. PMID- 14558748 TI - Cardiac acceleration as a marker of vagal withdrawal in heart rate control during exercise in humans. AB - OBJECTIVE: To investigate whether the time rate of change in heart rate i.e. cardiac acceleration, during aerobic exercise in human subjects could be used to differentiate vagal withdrawal from sympathetic stimulation. METHODS: Fifteen male subjects exercised on a bicycle ergometer at 50 Watts (Step 1), then 100 Watts (Step 2), for 2 minutes each. RESULTS: Heart rate (HR) was monitored from a resting value (mean +/- SD) of 80.3 +/- 12.9 to 113.8 +/- 13.6 beats min-1 in Step 1. In Step 2 exercise, HR increased from 113.8 +/- 13.6 to 145 +/- 20 beats min-1. At the initiation of Step 1, a rapid acceleration of HR was observed in the form of an overshoot response. In contrast to Step 1, a small overshoot response of cardiac acceleration was observed during Step 2. The difference between the mean cardiac acceleration at 10 seconds in Steps 1 and 2 was significant (2.40 +/- 0.19 and 0.71 +/- 0.12 beats min-1 sec-1, p<0.0001). CONCLUSION: The initial vagal withdrawal of exercise-induced tachycardia, as a frontline adaptive mechanism, can be indirectly identified from HR transients using cardiac acceleration as a new marker. PMID- 14558749 TI - Preparation of stable isotope-labeled 2-nitrobenzaldehyde derivatives of four metabolites of nitrofuran antibiotics and their comprehensive characterization by UV, MS, and NMR techniques. AB - A convenient method is presented for the preparation of the carbon-13-labeled 2 nitrobenzaldehyde derivatives of the nitrofuran metabolites 3-amino-2 oxazolidinone (AOZ), semicarbazide (SC), 1-aminohydantoin (AH), and 3-amino-5 morpholinomethyl-2-oxazolidinone (AMOZ), with the purpose of using them as internal standards for the quantification of trace levels of nitrofuran residues by liquid chromatography-tandem mass spectrometry in foods of animal origin. The synthesis encompasses the nitration of [1,2,3,4,5,6-(13)C(6)]toluene prior to chromyl compound-mediated oxidation of the methyl group into the corresponding aldehyde. The four metabolites of nitrofuran antibiotics were derivatized independently with the resulting ring-labeled 2-nitrobenzaldehyde (NBA) to obtain the target compounds. Both the isotopically enriched and native substances were used to perform a comprehensive fragmentation study by electrospray ionization (ESI) collision-induced dissociation (CID) mass spectrometry (MS). Full characterization of the nitrofuran derivatives was accomplished with ultraviolet (UV) and exhaustive nuclear magnetic resonance (NMR) analysis. A major advantage of the described procedure is that it can be extended to the preparation of other carbon-13-labeled derivatives of metabolites of nitrofuran antibiotics. PMID- 14558750 TI - Fast kinetic determination of 1-naphthylacetic acid in commercial formulations, soils, and fruit samples using stopped-flow phosphorimetry. AB - A kinetic method has been developed for the determination of 1-naphthylacetic acid by means of micellar-stabilized room temperature phosphorescence (MSRTP) using the stopped-flow mixing technique. The main feature of this system is that it diminishes the time required for the deoxygenation of the micellar medium and for the phosphorescence development. Phosphorescence enhancers such thallium(I) nitrate, sodium dodecyl sulfate (SDS), and sodium sulfite were optimized to obtain maximum sensitivity. The pH was also optimized as it strongly affects the luminescent properties of 1-naphthylacetic acid. A pH of 6.6 was selected as adequate for the phosphorescence development. The kinetic curve of 1 naphthylacetic acid phosphorescence was scanned at lambda(ex) = 278 nm and lambda(em) = 490 nm, and the maximum rate of phosphorescence was taken as the analytical signal. This was obtained by calculating the maximum slope of the curve in an interval of 3.6 s as it provided a good noise-to-signal ratio. This method permitted the determination of 1-naphthylacetic acid throughout a concentration range of 100-1800 ng mL(-1) with high precision (relative standard error = 0.91% and relative standard deviation = 2.30%; 1-naphthylacetic acid concentration = 800 ng mL(-1)). According to the Clayton criterion, the detection limit was 45 ng mL(-1). The same limit resulted in 39.3 ng mL(-1) when the error propagation theory was applied. The applicability of the method was successfully demonstrated by determining 1-naphthylacetic acid in different kind of samples, such as phytosanitary products, soils, pears, and apples. Recovery values not significantly different from the nominal content or the spiked amount were found for these determinations. PMID- 14558751 TI - Improved solid-phase extraction procedure in the analysis of paralytic shellfish poisoning toxins by liquid chromatography with fluorescence detection. AB - The analysis of shellfish extracts for the determination of paralytic shellfish poisoning (PSP) toxins by liquid chromatography with fluorescence detection repeatedly showed the presence of a compound suspected to interfere with gonyautoxin 4. The first aim of this study was to confirm by liquid chromatography coupled to tandem mass spectrometry that this compound was not gonyautoxin 4. The second part of this work was to improve a nonvolumetric C(18) solid-phase extraction (SPE) procedure to evaluate the removal of the interference associated with the recovery of PSP toxins. The cleanup procedure was modified into a volumetric SPE procedure and proved to efficiently and totally remove the interference while recovering from 78 to 85% of the PSP toxins available as commercial standards (saxitoxin, neosaxitoxin, gonyautoxins 1-4) and considered as major PSP toxins in human intoxication, with 85% recovery for gonyautoxin 4. The efficiency of this cleanup procedure was checked on shellfish extracts containing this interference and originating from France and Turkey. PMID- 14558752 TI - High-performance capillary electrophoresis with indirect UV detection for determination of alpha-galactosides in Leguminosae and Brassicaceae. AB - A rapid, easy, and reproducible capillary electrophoresis method for determination of raffinose family oligosaccharides (alpha-galactosides) was developed. Sucrose, raffinose, stachyose, verbascose, and ajugose were determined with indirect UV detection at moderate alkaline pH 9.2, using pyridine-2,6 dicarboxylic acid as background electrolyte in a sodium tetraborate buffer with added cetyltrimethylammonium bromide. The separation efficiency measured by the number of theoretical plates (N) ranged from 1.4 x 10(5) to 2.3 x 10(5). The precision of the method, measured by the relative standard deviation (RSD), was less than 0.53% for the migration times and better than 3.4% for normalized areas (NA), considering all sugars except verbascose (RSD(NA) = 11.8%). Detection limits were about 110 microg/mL, corresponding to 150-320 microM. Relative response factors (RRF) were calculated on the basis of linearity studies and used for quantification of alpha-galactosides in a lupine sample (Lupinus angustifolius). PMID- 14558753 TI - GC-MS quantitation of fragrance compounds suspected to cause skin reactions. 1. AB - Recent changes in European legislation require monitoring of 24 volatile compounds in perfumes as they might elicit skin sensitization. This paper reports a GC-MS quantitation procedure for their determination in fragrance concentrates. GC and MS conditions were optimized for a routine use: analysis within 30 min, solvent and internal standard selection, and stock solution stability. Calibration curves were linear in the range of 2-100 mg/L with coefficients of determination in excess of 0.99. The method was tested using real perfumes spiked with known amounts of reference compounds. PMID- 14558754 TI - Bruise detection in Pacific pink salmon (Oncorhynchus gorbuscha) by visible and short-wavelength near-infrared (SW-NIR) spectroscopy (600-1100 nm). AB - Visible and short-wavelength near-infrared (SW-NIR) spectroscopy (600-1100 nm) was used to detect bruises in intact, whole Pacific pink salmon (Oncorhynchus gorbuscha). The measurements were performed noninvasively through the skin and scales in the diffuse reflectance mode. Digital images of bruised and nonbruised regions of fish were captured after the fish samples were filleted. Image analysis was conducted using Adobe Photoshop 7.0 with relative gray values used as reference values in a partial least-squares (PLS) model. A PLS cross validation model using six latent variables yielded a standard error of prediction (SEP = 0.05%, R = 0.83). Approximately 84% of all nonbruised spectra were correctly classified, whereas approximately 81% of all bruised spectra were correctly classified. These results suggest that visible and SW-NIR could be used to control the bruise defect of fish products during processing, thereby improving product consistency and quality. PMID- 14558755 TI - Evaluation of the NO scavenging activity of procyanidin in grape seed by use of the TMA-PTIO/NOC 7 ESR system. AB - The nitrogen monoxide (NO) scavenging activity of grape seed extract (GSE) was studied in the TMA-PTIO/NOC 7 system. The procyanidin-rich (>95%) GSE showed strong NO scavenging activity in the system. The activity was found to depend on the condensation rate of cyanidin when synthetic oligomers were tested. Investigation of the NO scavenging activities of other polyphenols (catechin, epicatechin, epigallocatechin, and epigallocatechin gallate) in the TMA-PTIO/NOC 7 system revealed that gallocatechin, epigallocatechin, and epigallocatechin gallate exhibited strong activities. From the results, it was suggested that the high condensation rate of and the gallate ester moiety in procyanidin in GSE may play an important role in the NO scavenging activity. The mechanism of the NO scavenging activity of phenolic compounds such as GSE is speculated to be as follows: NO reacts with phenolic compounds directly to generate phenoxy radicals. PMID- 14558756 TI - Antimutagenic activity of phenylpropanoids from clove (Syzygium aromaticum). AB - Phenylpropanoids that possess antimutagenic activity were isolated from the buds of clove (Syzygium aromaticum). The isolated compounds suppressed the expression of the umu gene following the induction of SOS response in the Salmonella typhimurium TA1535/pSK1002 that have been treated with various mutagens. The suppressive compounds were mainly localized in the ethyl acetate extract fraction of the processed clove. This ethyl acetate fraction was further fractionated by silica gel column chromatography, which resulted in the purification and subsequent identification of the suppressive compounds. Electron impact mass spectrometry, IR, and (1)H and (13)C NMR spectroscopy were then used to delineate the structures of the compounds that confer the observed antimutagenic activity. The secondary suppressive compounds were identified as dehydrodieugenol (1) and trans-coniferyl aldehyde (2). When using 2-(2-furyl)-3-(5-nitro-2 furyl)acrylamide (furylfuramide) as the mutagen, compound 1 suppressed 58% of the umu gene expression as compared to the controls at a concentration of 0.60 micromol/mL, with an ID(50) (50% inhibitory dose) value of 0.48 micromol/mL, and compound 2 suppressed 63% of the umu gene expression as compared to the controls at a concentration of 1.20 micromol/mL, with an ID(50) value of 0.76 micromol/mL. Additionally, compounds 1 and 2 were tested for their ability to suppress the mutagenic activity of other well-known mutagens such as 4-nitroquinolin 1-oxide (4NQO) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), which do not require liver metabolizing enzymes, and aflatoxin B(1) (AfB(1)) and 3-amino-1,4-dimethyl 5H-pyrido[4,3-b]indole (Trp-P-1), which require liver metabolizing enzymes and activated Trp-P-1 and UV irradiation. Compounds 1 and 2 showed dramatic reductions in their mutagenic potential of all of the aforementioned chemicals or treatment. For the search of the structure-activity relationship, the derivatives of 1 and 2 (1a and 2a-c) were also assayed with all mutagens. Finally, the antimutagenic activities of compounds 1, 1a, 2, and 2a-c against furylfuramide, Trp-P-1, and activated Trp-P-1 were assayed by the Ames test using the S. typhimurium TA100 strain. PMID- 14558757 TI - Phenolic constituents and antioxidant properties of Xanthosoma violaceum leaves. AB - An extract of Xanthosoma violaceum leaves was subjected to a polyphenol profile determination, including total polyphenols, and antioxidant activity evaluation. Analysis of the extract resulted in the isolation of a new flavone C-glycoside, apigenin 6-C-beta-D-glucopyranosyl-8-C-beta-D-apiofuranoside (1), as well as known flavone C-glycosides, including vitexin (2), isovitexin (3), isovitexin 4' O-rhamnopyranoside (4), apigenin 6-C-[beta-D-glucopyranosyl-(1-->6)-beta-D glucopyranoside] (5), and apigenin 6,8-diC-beta-D-glucopyranoside (6). The antioxidant activity of the extract was assessed by means of two different in vitro tests: bleaching of the stable 1,1-diphenyl-2-picrylhydrazyl radical (DPPH test) and peroxidation induced by the water-soluble radical initiator 2,2' azobis(2-amidinopropane) hydrochloride, on mixed dipalmitoylphosphatidylcholine/linoleic acid unilamellar vesicles (LP-LUV test). In both tests used, the extract and a fraction II showed a significant antioxidant/free-radical scavenging effect (fraction II, EC(50) = 11.6 microg/mL) in comparison to alpha-tocopherol (EC(50) = 10.1 microg/mL). PMID- 14558758 TI - Biochemical outcome of blocking the ergot alkaloid pathway of a grass endophyte. AB - Neotyphodium sp. Lp1, an endophytic fungus from perennial ryegrass (Lolium perenne), produces the mycotoxin ergovaline in infected grasses, whereas a mutant in which a particular peptide synthetase gene is knocked out does not. We examined the impact of this knockout on other constituents of the ergot alkaloid pathway. Two simple lysergic acid amides, ergine and a previously undescribed amide, were eliminated by the knockout. Lysergic acid accumulated in the knockout endophyte, but quantities were only 13% of the total lysergic acid derivatives accumulated in the wild type. Concentrations of several clavines were not substantially affected. However, a novel clavine accumulated to higher concentrations in perennial ryegrass containing the knockout strain. The results indicate that production of simple lysergic acid amides requires the activity or products of the ergovaline-associated peptide synthetase and that the regulation of ergot alkaloid production is modified in response to the relatively late block in the pathway. PMID- 14558759 TI - Isolation and identification of steroidal saponins in Taiwanese yam cultivar (Dioscorea pseudojaponica Yamamoto). AB - A new furostanol pentaoligoside and spirostanol tetraoligoside were isolated for the first time from yam tubers (Dioscorea pseudojaponica Yamamoto) from Taiwan, together with four known yam saponins, methyl protodioscin, methyl protogracillin, dioscin, and gracillin. Their structures were characterized as 26 O-beta-D-glucopyranosyl-22alpha-methoxyl-(25R)-furost-5-en-3beta,26-diol, 3-O alpha-L-rhamnopyranosyl-(1-->2)-O-([alpha-L-rhamnopyranosyl-(1-->4)]-O-[alpha-L rhamnopyranosyl-(1-->4)])-beta-D-glucopyranoside, and (25R)-spirost-5-en-3beta-ol 3-O-alpha-L-rhamnopyranosyl-(1-->2)-O-([alpha-L-rhamnopyranosyl-(1-->4)]-O-[alpha L-rhamnopyranosyl-(1-->4)])-beta-D-glucopyranoside. The structural identification was performed using LC-MS and 1H and 13C NMR. The methanol extract of yam tubers was fractionated by XAD-2 column chromatography using a methanol/water gradient elution system to yield furostanol and spirostanol glycoside fractions. Preparative high-performance liquid chromatography, employing a C18 column and a mobile phase of methanol/water (69:31, v/v), was used to separate each furostanol glycoside, whereas a mobile phase of methanol/water (79:21, v/v) was used to resolve the individual spirostanol glycosides. The conversions from steroid saponins to diosgenin after acid hydrolysis were around 68 and 90% for furostanol and spirostanol glycosides, respectively. PMID- 14558760 TI - Antioxidant isoflavones in Osage orange, Maclura pomifera (Raf.) Schneid. AB - Recent findings that many human chronic diseases are associated with oxidative stresses have instigated the search for dietary antioxidants. Many phytochemicals, particularly phenolic compounds, have been found to possess strong antioxidant activity and reduce the risks of those diseases. Isoflavones, a special phenolic group found in soybean, have been found to act as antioxidants in some model systems. This study investigated the isoflavone content in a unique nonedible tree fruit, Osage orange [Maclura pomifera (Raf.) Schneid], and methods for the extraction, identification, and quantification of the two major isoflavones, osajin and pomiferin, were developed. The ethyl acetate extract contained 25.7% osajin and 36.2% pomiferin, and the two isoflavones were at 9.5 g kg(-1) of fresh Osage orange. Two model systems, FRAP and beta-CLAMS, were used to measure the antioxidant activity of these two isoflavones. Pomiferin was found to be a strong antioxidant in both systems, comparable to the antioxidant vitamins C and E and the synthetic antioxidant BHT. Osajin and the two soybean isoflavones (genistein and daidzein) showed no antioxidant activity. Although the Osage orange fruit is not a food source, it is considered to be safe and, therefore, a potentially good source of an antioxidant nutraceutical and functional food ingredient. PMID- 14558761 TI - Tartary buckwheat (Fagopyrum tataricum Gaertn.) as a source of dietary rutin and quercitrin. AB - Two samples of tartary buckwheat (Fagopyrum tataricum Gaertn.) from China and one from Luxembourg were studied by high-performance liquid chromatography (HPLC) to reveal the possibilities of growing tartary buckwheat herb as a possible source of rutin, quercetin, and quercitrin. The content of rutin was determined as up to 3% dry weight (DW) in tartary buckwheat herb. Quercitrin values were in the range of 0.01-0.05% DW. Only traces of quercetin were detected in just some of the samples. Tartary buckwheat seeds contained more rutin (about 0.8-1.7% DW) than common buckwheat seeds (0.01% DW). Rutin and quercetin content in seeds depends on variety and growing conditions. Tartary buckwheat seeds contained traces of quercitrin and quercetin, which were not found in common buckwheat seeds. PMID- 14558762 TI - Prenylated flavanones isolated from flowers of Azadirachta indica (the neem tree) as antimutagenic constituents against heterocyclic amines. AB - Four prenylated flavanones were isolated from the methanol extract of the flowers of Azadirachta indica (the neem tree) as potent antimutagens against Trp-P-1 (3 amino-1,4-dimethyl-5H-pyrido[4,3-b]indole) in the Salmonella typhimurium TA98 assay by activity-guided fractionation. Spectroscopic properties revealed that those compounds were 5,7,4'-trihydroxy-8-prenylflavanone (1), 5,4'-dihydroxy-7 methoxy-8-prenylflavanone (2), 5,7,4'-trihydroxy-3',8-diprenylflavanone (3), and 5,7,4'-trihydroxy-3',5'-diprenylflavanone (4). All isolated compounds were found for the first time in this plant. The antimutagenic IC(50) values of compounds 1 4 were 2.7 +/- 0.1, 3.7 +/- 0.1, 11.1 +/- 0.1, and 18.6 +/- 0.1 microM in the preincubation mixture, respectively. These compounds also similarly inhibited the mutagenicity of Trp-P-2 (3-amino-1-methyl-5H-pyrido[4,3-b]indole) and PhIP (2 amino-1-methyl-6-phenylimidazo[4,5-b]pyridine). All of the compounds 1-4 strongly inhibited ethoxyresorufin O-dealkylation activity of cytochrome P450 1A isoforms, which catalyze N-hydroxylation of heterocyclic amines. However, compounds 1-4 did not show significant inhibition against the direct-acting mutagen NaN(3). Thus, the antimutagenic effect of compounds 1-4 would be mainly based on the inhibition of the enzymatic activation of heterocyclic amines. PMID- 14558763 TI - Comparison of antioxidative properties of carbazole alkaloids from Murraya koenigii leaves. AB - A new dimeric carbazole alkaloid, 8,10'-[3,3',11,11'-tetrahydro-9,9'-dihydroxy 3,3',5,8'-tetramethyl-3,3'-bis(4-methyl-3-pentenyl)]bipyrano[3,2-a]carbazole (12), was isolated from the CH(2)Cl(2) extract of Murraya koenigii together with six known carbazole alkaloids, koenimbine (6), O-methylmurrayamine A (7), O methylmahanine (8), isomahanine (9), bismahanine (10), and bispyrayafoline (11). Their structures were determined on the basis of (1)H and (13)C NMR spectroscopic and mass spectrometric (MS) data. The antioxidative properties of 12 carbazole alkaloids isolated from leaves of M. koenigii were evaluated on the basis of the oil stability index together with their radical scavenging ability against 1,1 diphenyl-2-picrylhydrazyl (DPPH) radical. On the basis of the lag time to reach a steady state, the 12 carbazoles were classified into three groups. It is suggested that an aryl hydroxyl substituent on the carbazole rings plays a role in stabilizing the thermal oxidation and rate of reaction against DPPH radical. PMID- 14558764 TI - Physicochemical properties and bioactivity of nisin-containing cross-linked hydroxypropylmethylcellulose films. AB - Cross-linked hydroxypropylmethylcellulose (HPMC) cast films with citric acid as polycarboxylic cross-linker were elaborated to study the effect of cross-linking level on various properties. Increased amounts of cross-linking agent were not connected to statistically different tensile strength and Young's modulus. Whatever the cross-linking level of the film was, the ultimate elongation parameter decreased by approximately 60% compared to the HMPC control film. Moisture sorption isotherms and water contact angle meter showed that the effect of cross-linking degree tends to reduce the hygroscopic and hydrophilic characteristics of films. In addition, to control bacteria growth on food surfaces, the antimicrobial activity of both 98% cross-linked HPMC-nisin and control HPMC-nisin films was tested on Micrococcus luteus. Despite the incorporation of a significant content of nisin, cross-linked HPMC-nisin films were completely inactive on the microbial strain compared to the HPMC-nisin control films. Cross-linking conditions likely either denatured the nisin or irreversibly bound nisin to the cross-linked HPMC. However, nisin adsorbed into films made from previously cross-linked HPMC maintained its activity. PMID- 14558765 TI - Monomeric, oligomeric, and polymeric flavan-3-ol composition of wines and grapes from Vitis vinifera L. Cv. Graciano, Tempranillo, and Cabernet Sauvignon. AB - The monomeric, oligomeric, and polymeric flavan-3-ol composition of wines, grape seeds, and skins from Vitis vinifera L. cv. Graciano, Tempranillo, and Cabernet Sauvignon has been studied using (1) fractionation by polyamide column chromatography followed by HPLC/ESI-MS analysis, (2) fractionation on C(18) Sep Pak cartridges followed by reaction with vanillin and acid-catalyzed degradation in the presence of toluene-alpha-thiol (thiolysis). The content of monomers ((+) catechin and (-)-epicatechin), procyanidin dimers (B3, B1, B4, and B2), trimers (T2 and C1), and dimer gallates (B2-3-O-gallate, B2-3'-O-gallate, and B1-3-O gallate) ranged from 76.93 to 133.18 mg/L in wines, from 2.30 to 8.21 mg/g in grape seeds, and from 0.14 to 0.38 mg/g in grape skins. In wines, the polymeric fraction represented 77-84% of total flavan-3-ols and showed a mean degree of polymerization (mDP) value of 6.3-13.0. In grapes, the polymeric fraction represented 75-81% of total flavan-3-ols in seeds and 94-98% in skins and showed mDP values of 6.4-7.3 in seeds and 33.8-85.7 in skins. All the monomeric flavan-3 ols and oligomeric procyanidins found in wines were also present in seeds, although differences in their relative abundances were seen. The skin polymeric proanthocyanidins participated in the equilibration of the wine polymeric proanthocyanidin fraction, especially contributing to the polymer subunit composition and mDP. PMID- 14558766 TI - Changes in the polyphenolic and volatile contents of "fino" sherry wine exposed to ultraviolet and visible radiation during storage. AB - Experiments of accelerated oxidation of "fino" sherry wines have been conducted at 25 degrees C and under the influence of UV-visible radiation (a xenon lamp of 1500 W). With the aim of determining the contribution of UV-vis radiation to the browning phenomenon, two types of glass bottles were employed: topaz bottles (with low values of transmittance in the UV-visible range) and transparent bottles. To identify significant differences between the wine before and after being subjected to the influence of the UV-vis radiation, the values of absorbance at 420 nm and the concentrations of various polyphenolic and volatile compounds were submitted to a multivariate variance analysis. Both factors considered (time and type of bottle) had a statistically significant effect on the values of absorbance at 420 nm and on the concentration of most of the polyphenolic compounds, whereas only the "time" factor was significant for volatile compounds. All wines showed losses in several polyphenolic compounds, which were more severe for the wines bottled in transparent glasses. However, these wines exhibited a lower degree of visual browning (abs 420 nm). In the case of volatile compounds, most of these presented increases during storage exposed to the influence of the UV-vis radiation. PMID- 14558767 TI - Rheological properties of concentrated skim milk: influence of heat treatment and genetic variants on the changes in viscosity during storage. AB - Heat treatment during manufacturing of milk powder is one of the most important tools for manipulation of its functional properties, and it is the basis of the classification of these proteins into low-, medium-, and high-heat types. Slight differences in the sequences of the major proteins in milk (genetic variants) seem to have also a significant effect in milk powder processing (U.S. patent). Therefore, the effects of high-temperature storage and heat treatment on skim milk of defined genetic variants of beta-lactoglobulin (beta-LG) were measured. The samples had 45% total solids, the temperature of aging was 50 degrees C, and the heat treatment was 90 degrees C for 10 min prior to evaporation. Measurements on shear rate and on apparent viscosity were determined for each sample. During storage of the concentrated milk, the apparent viscosity and yield values increased markedly, and the age-dependent increase in viscosity in heat-treated concentrated skim milks was much more pronounced than in those prepared from unheated skim milks. The increase in apparent viscosity and yield value with storage time was notably different for milks containing different genetic variants. Unheated concentrated milks containing the B variant of beta-LG showed the most rapid increase in apparent viscosity with storage time, whereas the viscosity increase was slowest in the concentrate containing the A variant. In contrast, heat-treated concentrated milks containing the A variant of beta-LG showed the most rapid increase in viscosity with storage time, whereas the viscosity increase was slowest in the concentrate containing the AB variant. The changes in apparent viscosity of concentrated milk were largely reversible under high shear during the early stages of storage, but samples stored for a long time showed irreversible changes in apparent viscosity. Particle size analysis confirmed irreversible aggregation and fusion of casein particles during storage. PMID- 14558768 TI - Discrimination between arabica and robusta coffee species on the basis of their amino acid enantiomers. AB - This work reports the results for the composition of robusta and arabica coffee species in terms of their amino acid enantiomers in the green and roasted states. The analyses were conducted for the free amino acids, as well as for the amino acids obtained after acid hydrolysis. The amino acids were extracted/hydrolyzed and isolated by SPE on strong cation exchange columns, derivatized to their N ethoxycarbonylheptafluorobutyl esters, and analyzed by gas chromatography/FID on a Chirasil l-Val column. Multivariate analyses applied to the results showed that the free amino acids can be used as a tool for discrimination between coffee species, with a special reference to l-glutamic acid, l-tryptophan, and pipecolic acid. There is also some evidence that these compounds can be used for discrimination between green coffees subjected to different postharvest processes. It is also shown that the amino acid levels observed after acid hydrolysis can be used for the same purposes, although displaying less discriminatory power. PMID- 14558769 TI - Effect of gamma-irradiation on the lipid profile of nutmeg (Myristica fragrans Houtt.). AB - The effect of gamma-irradiation on the lipid constituents of nutmeg (Myristica fragrans) was examined at radiation doses between 2.5 and 10 kGy. The fatty acid composition of the triacylglycerol, the major lipid component, was found to be made up of myristic (90%), palmitic (6%), lauric (3%), petroselinic (0.13%), and stearic acids (0.5%) as determined by gas chromatography-mass spectrometry. A dose-dependent decrease in the triacylglycerol content and a concomitant increase in free fatty acids characterized the lipid profile of the irradiated spice. This suggested a breakdown of acylglycerols during radiation processing, resulting in the release of free fatty acids. These changes were found to be significant at doses above 5 kGy. The impact of the above changes on the flavor of the spice is discussed. These studies suggest that radiation processing of nutmeg should be limited to a dose of 5 kGy. PMID- 14558770 TI - Effect of nitrogen concentration of the nutrient solution on the volatile constituents of leaves of Salvia fruticosa Mill. in solution culture. AB - Essential oils from hydroponically cultivated Salvia fruticosa were analyzed by GC-MS techniques. Three different levels of nitrogen (100, 150, and 200 mg/L) were used in the nutrient solution for the cultivation, using the nutrient film technique. A total of 79 compounds were identified, and qualitative and quantitative differences have been observed between the samples collected at full bloom (flowering stage) and at the end of the seed formation stage. 1,8-Cineole, beta-caryophyllene, and viridiflorol were the predominant constituents in most cases. 13-epi-Manool was identified by using GC parameters and mass spectrum fragmentation pattern, whereas labd-7,13-dien-15-ol, a labdane type diterpene, was identified for the first time in the genus Salvia, using GC parameters and an authentic sample. The results obtained from GC-MS analyses were submitted to chemometric analysis. PMID- 14558771 TI - Quantification of polyphenolics and their antioxidant capacity in fresh plums. AB - Total phenolics, total flavonoids, and antioxidant capacity of 11 cultivars of fresh plums were determined using spectrophotometric methods. Identification and quantification of individual polyphenolics were performed using reversed-phase high-performance liquid chromatography equipped with a diode array detector. The total phenolic contents of various cultivars widely varied from 125.0 to 372.6 mg/100 g expressed as gallic acid equivalents. The level of total flavonoids in fresh plums ranged between 64.8 and 257.5 mg/100 g expressed as catechin equivalents. Antioxidant capacity, expressed as vitamin C equivalent antioxidant capacity (VCEAC), ranged from 204.9 to 567.0 mg/100 g with an average of 290.9 mg/100 g of fresh weight. Cv. Beltsville Elite B70197 showed the highest amounts of total phenolics and total flavonoids and the highest VCEAC. A positive relationship (correlation coefficient r (2)() = 0.977) was presented between total phenolics and VCEAC, suggesting polyphenolics would play an important role in free radical scavenging. The level of IC(50) value of superoxide radical anion scavenging activity of the plum cultivars ranged from 13.4 to 45.7 mg of VCEAC/100 g. Neochlorogenic acid was the predominant polyphenolic among fresh plums tested. Flavonols found in plum were commonly quercetin derivatives. Rutin was the most predominant flavonol in plums. Various anthocyanins containing cyanidin aglycon and peonidin aglycon were commonly found in all plums except for cv. Mirabellier and NY 101. PMID- 14558773 TI - Soil amino acid composition quantified by acid hydrolysis and anion chromatography-pulsed amperometry. AB - Soil organic N accounts for 95-98% of the total soil N content with amino acids (AAs) and amino sugars (ASs) identified as the major soil organic N compounds, but traditional 6 M HCl with reflux or sealed digestions for 24 h and various detection systems have accounted for only 30-40% of soil total N content as AA-N. This study compared traditional HCl extraction methodology with methanesulfonic acid (MSA) hydrolysis and nonderivatized AA and AS quantification by ion chromatography with pulsed amperometric detection for determination of the AA composition of plant litter and soils. MSA (4 M) gave AA-N recovery comparable to or better than 6 M HCl for plant AA digestions (16 h, 121 degrees C, 104 kPa). Use of 4 M MSA (0.5-1.5 h, 136 degrees C, 112 kPa) increased the total recovery of organic N as AAs, ASs, and NH(4)(+) by 46% from soils (n = 22) compared with 6 M HCl (12 h, 110 degrees C, reflux) with a MSA recovery rate of 85.6% of the total N content (n = 22 soils). The shorter MSA soil digestions (0.5-1.5 h) suggested that the majority of soil organic N was not present as protein forms found in plant litter analysis (16 h of digestion). MSA ion chromatographic analysis for soil AA/AS composition is a robust nonderivatization method requiring little sample preparation that can distinguish between small changes in soil AA composition during one growing season due to vegetation and tillage managements. PMID- 14558772 TI - Major phenolics in apple and their contribution to the total antioxidant capacity. AB - The contribution of each phytochemical to the total antioxidant capacity of apples was determined. Major phenolic phytochemicals of six apple cultivars were identified and quantified, and their contributions to total antioxidant activity of apples were determined using a 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging assay and expressed as vitamin C equivalent antioxidant capacity (VCEAC). Average concentrations of major phenolics and vitamin C in six apple cultivars were as follows (mg/100 g of fresh weight of apples): quercetin glycosides, 13.20; procyanidin B(2), 9.35; chlorogenic acid, 9.02; epicatechin, 8.65; phloretin glycosides, 5.59; vitamin C, 12.80. A highly linear relationship (r (2) > 0.97) was attained between concentrations and total antioxidant capacity of phenolics and vitamin C. Relative VCEAC values of these compounds were in the order quercetin (3.06) > epicatechin (2.67) > procyanidin B(2) (2.36) > phloretin (1.63) > vitamin C (1.00) > chlorogenic acid (0.97). Therefore, the estimated contribution of major phenolics and vitamin C to the total antioxidant capacity of 100 g of fresh apples is as follows: quercetin (40.39 VCEAC) > epicatechin (23.10) > procyanidin B(2) (22.07) > vitamin C (12.80) > phloretin (9.11) > chlorogenic acid (8.75). These results indicate that flavonoids such as quercetin, epicatechin, and procyanidin B(2) rather than vitamin C contribute significantly to the total antioxidant activity of apples. PMID- 14558774 TI - Volatile constituents of uncooked rhubarb (Rheum rhabarbarum L.) stalks. AB - Volatiles of rhubarb (Rheum rhabarbarum L.) stalks were isolated by means of vacuum headspace method and analyzed by capillary gas chromatography-mass spectrometry. Fifty-nine components were reported for the first time in rhubarb. A striking feature of the extracts obtained is the preponderance (approximately 65%) of compounds with C(6) skeletons. In addition to unsaturated C(6) aldehydes and alcohols, substantial amounts of the less common (E)-2- and (E)-3-hexenoic acid were detected. Gas chromatography-olfactometry and determination of odor activity values revealed the sensory importance of the C(6) compounds to the aroma of rhubarb. Comparative experiments involving the inhibition of enzyme activities revealed that the initial spectrum of C(6) components is changed due to subsequent isomerization and reductions. Thus, contributions of (Z)-3-hexenal and the unsaturated acids decrease, and (E)-2-hexenal/(E)-2-hexenol play major sensory roles. PMID- 14558775 TI - Monitoring carbonyl-amine reaction between pyruvic acid and alpha-amino alcohols by FTIR spectroscopy--a possible route to Amadori products. AB - The carbonyl-amine reaction between pyruvic acid and alpha-amino alcohols was monitored by Fourier transform infrared spectroscopy at a temperature range between 20 and 100 degrees C and under acidic and basic conditions. To avoid interference, the reactions were conducted in the absence of solvent using liquid reactants such as methyl pyruvate, pyruvic acid, ethanolamine, and 1-amino-2,3 propanediol. Analysis of the time- and temperature-dependent spectra indicated that under basic conditions and at room temperature, the initial imine formation and its subsequent isomerization through a 1,3-prototropic shift occur very rapidly and the reaction goes to completion within 12 min. Interestingly, the isomerization product of the initial imine is the so-called Schiff base intermediate formed when the corresponding amino acid and the reducing sugar react during a typical Maillard reaction. Furthermore, the detailed studies also indicated that during the first 30 s, the rate of formation of the initial imine was faster than the rate of its isomerization; however, after 60 s, its rate of isomerization becomes faster than the rate of its formation. The data also indicated that under acidic conditions, this isomerization was prevented from occurring and the reaction was terminated at the initial imine formation stage. In addition, temperature-dependent spectra indicated that the isomerization of the Schiff's base into eneaminol can be achieved at or above 60 degrees C and its subsequent rearrangement into Amadori product can be attained at temperatures above 80 degrees C even under basic conditions, thus providing a novel route to Maillard reaction products starting from a keto acid and an amino alcohol. This observation was also confirmed through identification of the common Amadori product in both keto acid/amino alcohol and sugar/amino acid mixtures, by the application of tandem mass spectrometry and chemical ionization techniques. PMID- 14558776 TI - Modification of volatile compound profile of virgin olive oil due to hot-water treatment of olive fruit. AB - The effect of hot-water treatments of olive fruits before processing on the biosynthesis of virgin olive oil aroma was investigated by quantifying the variation within the major classes of volatile compounds. Data showed that hot water treatments gave rise to changes in the volatile aroma profile of virgin olive oil from the three olive cultivars under study, Manzanilla, Picual, and Verdial. Different effects by thermal treatments were observed according to cultivar. In general, these changes are mainly due to a decrease in the contents of C(6) aldehydes and C(5) compounds. Contents of C(6) alcohols and esters remained constant or decreased slightly when the temperature of the treatment was increased. Thus, heat treatments seemed to promote a partial deactivation of the lipoxygenase/hydroperoxide lyase enzyme system, whereas other enzymatic activities, within the lipoxygenase pathway, such as alcohol dehydrogenase and alcohol acyltransferase, remained apparently unaffected as a consequence of heat treatments. PMID- 14558777 TI - Effect of sucrose ester addition on nucleation and growth behavior of milk fat sunflower oil blends. AB - The effects of addition of the sucrose esters (SE) P-1670, P-170, and S-170 to a high-melting fraction of milk fat (HMF) and its blends with sunflower oil (SFO) on nucleation and growth were studied by laser polarized light turbidimetry and polarized light microscopy (PLM). The three SE delayed nucleation of HMF at the temperatures selected. P-1670 did not modify average crystal size after 3 h at crystallization temperature (T(c)) or crystal size distribution and modified crystallization kinetics only slightly. P-170 and S-170, however, markedly diminished crystal size and narrowed crystal size distribution. Activation free energies of nucleation at equivalent supercooling, calculated using the Fisher Turnbull equation, significantly increased with addition of SE. According to these results, among the mechanisms described in the literature for fats or emulsions, the cocrystallization hypothesis is the one that better described the effects of sucrose esters on crystallization behavior in these systems. PMID- 14558778 TI - Analytical characterization of Moringa oleifera seed oil grown in temperate regions of Pakistan. AB - The hexane-extracted oil content of Moringa oleifera seeds ranged from 38.00 to 42.00%. Protein, fiber, and ash contents were found to be 26.50-32.00, 5.80-9.29, and, 5.60-7.50%, respectively. Results of physical and chemical parameters of the extracted oil were as follows: iodine value, 68.00-71.80; refractive index (40 degrees C), 1.4590-1.4625; density (24 degrees C), 0.9036-0.9080 mg/mL; saponification value, 180.60-190.50; unsaponifiable matter, 0.70-1.10%; and color (1 in. cell), 0.95-1.10 R + 20.00-35.30 Y. Tocopherols (alpha, gamma, and delta) in the oil were up to 123.50-161.30, 84.07-104.00, and 41.00-56.00 mg/kg, respectively. The oil was found to contain high levels of oleic acid (up to 78.59%) followed by palmitic, stearic, behenic, and arachidic acid up to levels of 7.00, 7.50, 5.99, and 4.21%, respectively. The induction period (Rancimat, 20 L/h, 120 degrees C) of the crude oil was 9.99 h and reduced to 8.63 h after degumming. Specific extinctions at 232 and 270 nm were 1.70 and 0.31, respectively. Many parameters of M. oleifera oil indigenous to Pakistan were comparable to those of typical Moringa seed oils reported in the literature. The results of the present analytical study were also compared with those of different vegetable oils. PMID- 14558779 TI - Solid-phase microextraction in the analysis of virgin olive oil volatile fraction: modifications induced by oxidation and suitable markers of oxidative status. AB - Modifications of virgin olive oil subjected to accelerated storage were evaluated by HS-SPME analysis. To find a suitable marker of oxidative degradation, the volatile compounds showing variable concentration during the oxidative process have been identified and quantified by SPME coupled to GC-MS and GC-FID, respectively. The SPME analysis results were then compared with the parameters usually applied to assess the oxidative status of lipids, such as peroxide value, spectrophotometric absorbance, and loss of unsaturated fatty acids. Finally, the assessment of nonanal has been suggested as a marker of oxidative degradation. This rapid, inexpensive, and reliable method may allow screening of oils prior to testing by a panel of assessors. PMID- 14558780 TI - Solid-phase microextraction in the analysis of virgin olive oil volatile fraction: characterization of virgin olive oils from two distinct geographical areas of northern Italy. AB - SPME was employed to characterize the volatile profile of virgin olive oils produced in two geographical areas of northern Italy: the region of the Gulf of Trieste and the area near Lake Garda. There are as yet no data on the headspace composition of virgin olive oils from these regions, characterized by particular conditions of growth for Olea europaea. Using the SPME technique coupled to GC-MS and GC-FID, the volatile components of 42 industrially produced virgin olive oil samples were identified and the principal compounds quantitatively analyzed. Significant differences in the proportion of volatile constituents from oils of different varieties and geographical origins were detected. The results suggest that besides the genetic factor, environmental conditions influence the volatile formation. PMID- 14558781 TI - Fermentation characteristics of some assamica clones and process optimization of black tea manufacturing. AB - Changes in the specific activities of polyphenol oxidase (PPO), peroxidase (POD), and protease and in the relative amounts of flavan-3-ols for eight genetically derived cultivated teas at various stages of leaf maturity and in four succescive seasons were examined. A series of investigations were carried out to study the cross-reactivity of complex polyphenols and PPO-generated orange-yellow theaflavins, as well as of POD oxidized substrates, producing brown so-called thearubigins during fermented tea processing. From the estimation of five major catechins, PPO activities in young shoots, and theaflavin and thearubigin contents of crushed, torn, and curled (CTC) black teas, the superior variety and flavorful flush characteristics were refined. Notable protein hydrolysis by endogenous protease as measured from free amino acids and formation of tannin protein complex (browning products) was obtained for cultivar character and product quality. Results showed that process optimization with respect to time, temperature, moisture, and pH maximizes PPO-catalyzed desirable theaflavin pigments, whereas POD-mediated chemical reaction produces dull color. PMID- 14558782 TI - In vivo studies on the metabolism of the monoterpene pulegone in humans using the metabolism of ingestion-correlated amounts (MICA) approach: explanation for the toxicity differences between (S)-(-)- and (R)-(+)-pulegone. AB - The major in vivo metabolites of (S)-(-)-pulegone in humans using a metabolism of ingestion-correlated amounts (MICA) experiment were newly identified as 2-(2 hydroxy-1-methylethyl)-5-methylcyclohexanone (8-hydroxymenthone, M1), 3-hydroxy-3 methyl-6-(1-methylethyl)cyclohexanone (1-hydroxymenthone, M2), 3-methyl-6-(1 methylethyl)cyclohexanol (menthol), and E-2-(2-hydroxy-1-methylethylidene)-5 methylcyclohexanone (10-hydroxypulegone, M4) on the basis of mass spectrometric analysis in combination with syntheses and NMR experiments. Minor metabolites were be identified as 3-methyl-6-(1-methylethyl)-2-cyclohexenone (piperitone, M5) and alpha,alpha,4-trimethyl-1-cyclohexene-1-methanol (3-p-menthen-8-ol, M6). Menthofuran was not a major metabolite of pulegone and is most probably an artifact formed during workup from known (M4) and/or unknown precursors. The differences in toxicity between (S)-(-)- and (R)-(+)-pulegone can be explained by the strongly diminished ability for enzymatic reduction of the double bond in (R) (+)-pulegone. This might lead to further oxidative metabolism of 10 hydroxypulegone (M4) and the formation of further currently undetected metabolites that might account for the observed hepatotoxic and pneumotoxic activity in humans. PMID- 14558783 TI - Comparative structure and physicochemical properties of Ilpumbyeo, a high-quality japonica rice, and its mutant, Suweon 464. AB - A new rice mutant Suweon 464 (S-464) derived from a high-quality rice, Ilpumbyeo (IP), revealed a striking difference in cooking quality from IP. The physicochemical properties of S-464 and IP were compared. S-464 was unusually high in amylose and fiber contents, had B-type crystallinity of starch, and had a markedly lower proportion of short chains in the distribution of glucan-chain fractions of debranched starch as compared with IP. Scanning electron microscopy revealed that starch granules of S-464 were much smaller in size than those of IP and that many of them were not separated from amyloplasts. The physicochemical properties of S-464 would contribute to poor gelatinization, lower swelling power, higher hardness, and less stickiness when cooked. Although S-464 may not be desirable for cooked rice, the mutant could be an excellent candidate for other processed food products on the basis of its unusual properties of starch and high fiber, protein, and lipid contents. PMID- 14558784 TI - Comparison of yield, composition, and antioxidant activity of turmeric (Curcuma longa L.) extracts obtained using various techniques. AB - Turmeric extracts were obtained from two lots of raw material (M and S) using various techniques: hydrodistillation, low pressure solvent extraction, Soxhlet, and supercritical extraction using carbon dioxide and cosolvents. The solvents and cosolvents tested were ethanol, isopropyl alcohol, and their mixture in equal proportions. The composition of the extracts was determined by gas chromatography flame ionization detection (GC-FID) and UV. The largest yield (27%, weight) was obtained in the Soxhlet extraction (turmeric (S), ethanol = 1:100); the lowest yield was detected in the hydrodistillation process (2.1%). For the supercritical extraction, the best cosolvent was a mixture of ethanol and isopropyl alcohol. Sixty percent of the light fraction of the extracts consisted of ar-turmerone, (Z)-gamma-atlantone, and (E)-gamma-atlantone, except for the Soxhlet extracts (1:100, ethanol), for which only ar-turmeronol and (Z)-alpha-atlantone were detected. The maximum amount of curcuminoids (8.43%) was obtained using Soxhlet extraction (ethanol/isopropyl alcohol). The Soxhlet and low pressure extract exhibited the strongest antioxidant activities. PMID- 14558785 TI - Production and characterization of oil-in-water emulsions containing droplets stabilized by beta-lactoglobulin-pectin membranes. AB - Oil-in-water emulsions containing droplets stabilized by beta-lactoglobulin (beta Lg)-pectin membranes were produced using a two-stage process. A primary emulsion containing small droplets (d(32) approximately 0.3 microm) was prepared by homogenizing 10 wt % corn oil with 90 wt % aqueous solution (1 wt % beta-Lg, 5 mM imidazole/acetate buffer, pH 3.0) using a high-pressure valve homogenizer. The primary emulsion was then diluted with pectin solutions to produce secondary emulsions with a range of pectin concentrations (5 wt % corn oil, 0.45 wt % beta Lg, 5 mM imidazole/acetate buffer, 0-0.22 wt % pectin, pH 3.0). The electrical charge on the droplets in the secondary emulsions decreased from +33 +/- 3 to -19 +/- 1 mV as the pectin concentration was increased from 0 to 0.22 wt %, which indicated that pectin adsorbed to the droplet surfaces. The mean particle diameter of the secondary emulsions was small (d(32) < 1 microm) at relatively low pectin concentrations (<0.04 wt %), but increased dramatically at higher pectin concentrations (e.g., d(32) approximately 13 microm at 0.1 wt % pectin), which was attributed to charge neutralization and bridging flocculation effects. Emulsions with relatively small mean particle diameters (d(32) approximately 1.2 microm at 0.1 wt % pectin) could be produced by disrupting flocs formed in secondary emulsions containing highly negatively charged droplets, for example, by sonication, blending, or homogenization. The particles in these emulsions probably consisted of small flocs containing a number of protein-coated droplets bound together by pectin molecules. These emulsions had good stability to further particle aggregation up to relatively high ionic strengths (< or =500 mM NaCl) and low pH (pH 3). The interfacial engineering technology used in this study could lead to the creation of food emulsions with improved physicochemical properties or stability. PMID- 14558787 TI - In vitro hypoglycemic effects of different insoluble fiber-rich fractions prepared from the peel of Citrus sinensis L. cv. Liucheng. AB - Insoluble fiber-rich fractions (FRFs), including insoluble dietary fiber, alcohol insoluble solid, and water-insoluble solid, were isolated from the peel of Citrus sinensis L. cv. Liucheng. We found that these three FRFs could effectively adsorb glucose, retard glucose diffusion, and inhibit the activity of alpha-amylase to different extents. These mechanisms might create a concerted benefit in decreasing the rate of glucose absorption and eventually lower the concentration of postprandial serum glucose. The potential hypoglycemic effects of these FRFs suggested that they could be incorporated as low-calorie bulk ingredients in high fiber foods to reduce calorie level and control blood glucose level. PMID- 14558786 TI - Metabolism of stevioside and rebaudioside A from Stevia rebaudiana extracts by human microflora. AB - Stevia rebaudiana standardized extracts (SSEs) are used as natural sweeteners or dietary supplements in different countries for their content of stevioside or rebaudioside A. These compounds possess up to 250 times the sweetness intensity of sucrose, and they are noncaloric and noncariogenic sweeteners. The aim of this study was to investigate the in vitro transformation of stevioside and rebaudioside A after incubation with human microflora, the influence of these sweeteners on human microbial fecal community and which specific groups metabolize preferentially stevioside and rebaudioside A. The experiments were carried out under strict anaerobic conditions in batch cultures inoculated with mixed fecal bacteria from volunteers. The hydrolysis was monitored by HPLC coupled to photodiode array and mass spectrometric detectors. Isolated bacterial strains from fecal materials incubated in selective broths were added to stevioside and rebaudioside A. These sweeteners were completely hydrolyzed to their aglycon steviol in 10 and 24 h, respectively. Interestingly, the human intestinal microflora was not able to degrade steviol. Furthermore, stevioside and rebaudioside A did not significantly influence the composition of fecal cultures; among the selected intestinal groups, bacteroides were the most efficient in hydrolyzing Stevia sweeteners to steviol. PMID- 14558788 TI - A combination of tea (Camellia senensis) catechins is required for optimal inhibition of induced CYP1A expression by green tea extract. AB - It was previously demonstrated that the commercial green tea extract Polyphenon 100 (P100), and to a lesser extent (-)-epigallocatechin-3-gallate (EGCG), partially antagonizes 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced transcription of human CYP1A1 (Williams, S. N.; Shih, H.; Guenette, D. K.; Brackney, W.; Denison, M. S.; Pickwell, G. V.; Quattrochi, L. C. Chem.-Biol. Interact. 2000, 128, 211-229). Here, P100 is compared to a reconstituted mixture of the four major tea catechins (referred to as P100R) to determine whether inhibition was due to additional polyphenols in the extract or from synergistic interactions among the tea catechins. It was found that cotreatment of cells with TCDD and either P100 or P100R inhibited TCDD-induced CYP1A promoter-driven luciferase reporter activity (HepG2 cells) and CYP1A expression (HepG2 and primary human hepatocytes), similarly. These results indicate that modulation of human CYP1A expression by P100 can be attributed entirely to the combination of the four tea catechins. These findings may be important in the evaluation of future chemoprevention strategies using green tea and single catechin agents. PMID- 14558789 TI - Protein surface recognition by rational design: nanomolar ligands for potassium channels. AB - The rational design and development of four-fold symmetrical ligands for potassium channels is described. PMID- 14558790 TI - Distinguishing rate-limiting electron versus H-atom transfers in Cu2O2-mediated oxidative N-dealkylations: application of inter- versus intramolecular kinetic isotope effects. AB - Copper-dioxygen adducts are important biological oxidants. To gain a better understanding of the underlying chemistries of such species, we report on a series of Cu2II-O2 complexes, [{CuII(MePY2)R'}2(O2)](B(C6F5)4)2 (1R') (where (MePY2)R' is a 4-pyridyl substituted bis[2-(2-(4-R'-pyridyl)ethyl]methylamine; R' = H, MeO, Me2N; Zhang, C. X.; et al. J. Am. Chem. Soc. 2003, 125, 634-635), which readily oxidize exogenous substrates. In this study, we explore the mechanism by which 1R' facilitates the oxidative N-dealkylation of para-substituted N,N dimethylanilines (R-DMA; R = MeO, Me, H, CN). In the case of 1H, the linear free energy correlation plot (rho = -2.1) and intramolecular deuterium kinetic isotope effect (KIEintra, using p-R-(C6H4)-N(CH3)(CD3)) profile suggest that R-DMA oxidation occurs through rate-limiting electron transfer (ET). This mechanism was further enforced by comparison of KIEintra versus the intermolecular KIE (KIEinter, using p-R-(C6H4)-N(CH3)2 versus p-R-(C6H4)-N(CD3)2). It was found that KIEinter < KIEintra, suggesting an ET process. In the case of both 1MeO and 1Me2N, the KIEintra profile and linear free-energy correlation plots (rho = -0.49 and -0.99 for 1Me2N and 1MeO with especially poor fitting for the latter) are inconclusive in distinguishing between a rate-limiting ET or hydrogen atom transfer (HAT) pathway. Comparisons of KIEinter versus KIEintra demonstrate a switch in mechanism from ET to HAT for 1Me2N and 1MeO oxidation of R-DMA as R-DMA is made less reducing. In the case of 1Me2N, MeO-DMA and Me-DMA are oxidized via a rate-limiting ET (KIEinter < KIEintra), while H-DMA and CN-DMA are oxidized through a HAT pathway (KIEinter approximately KIEintra). For 1MeO, oxidation occurs through an ET pathway for MeO-, Me-, and H-DMA (KIEinter < KIEintra), while CN-DMA is oxidized though a HAT process (KIEinter approximately KIEintra). Copper complex attributes, which may contribute to the mechanistic observations, are suggested. PMID- 14558791 TI - Enantioselective Michael reaction of malonates to nitroolefins catalyzed by bifunctional organocatalysts. AB - Michael reaction of malonates to nitroolefins with chiral bifunctional organocatalysts, bearing both a thiourea and tertiary amino group, afforded Michael adducts with high yields and enantioselectivities (up to 95%, up to 93% ee). PMID- 14558792 TI - C-H bond activation by unsymmetrical 2-(N-arylimino)pyrrolide Pt complexes: geometric effects on reactivity. AB - Reactions of chloroplatinum methyl complexes with N-(arylimino)pyrrolide anions afford cis and trans neutral platinum methyl complexes. Isomers with methyl trans to the pyrrolide nitrogen activate benzene C-H bonds at 85 degrees C more than 80 times faster than the corresponding cis isomer. In addition, reactions of platinum dimethyl complexes with N-(arylimino)pyrroles (Ar = 4-substituted phenyl) in C6D6 at ambient temperature give unlabeled methane and cis methyl complex containing heavily deuterated Pt-Me. In contrast, bulky aryl substituents give methane isotopomers and trans-Pt-Ph product. The origins of these observations are discussed. PMID- 14558793 TI - Regioselective bond cleavage in the dissociative electron transfer to benzyl thiocyanates. AB - The electrochemical reduction of benzyl thiocyanate and p-nitrobenzyl thiocyanate was investigated in acetonitrile at an inert electrode. These two compounds reveal a change in the reductive cleavage mechanism, and more interestingly, they show a clear-cut example of a regioselective bond dissociation. Both phenomena may be understood on the basis of the dissociative ET theory and its extension to the formation/dissociation reactions of radical ions. While the effect of the standard oxidation potential of the leaving group seems to be predominant in understanding the change in the ET mechanism by changing the driving force, the regioselective cleavage is dictated by changes in the intrinsic barrier related to the nature of the substituent on the aryl moiety. PMID- 14558794 TI - Formation and isolation of an iron-tripyrrole complex from heme degradation. AB - Addition of dioxygen to a solution of (py)2Fe(OEPO) (1) (where OEPO is the trianion of octaethyloxophlorin) in pyridine-d5 at 23 degrees C results in a series of oxidative changes that have been followed by 1H NMR spectroscopy. After 4 h, the resonances of 1 have vanished, while several sets of new resonances have developed. The most intense of these are identified as belonging to (py)2FeIII(OEB) (2) by comparison with the previously obtained spectrum of this species, which contains a ring-opened tetrapyrrole. After the sample was left standing for 1 day, further changes occur which produce a sample that is stable, because 1H NMR spectroscopic examination reveals no further changes on continued exposure to dioxygen or air. The product, (py)2FeIII(HETP) (3), where HETP is the dianion of a hexaethyltripyrrole, has been converted into (n-Bu4N)[ClFeIII(HETP)] by treatment with tetra(n-butyl)ammonium chloride. Red (n-Bu4N)[ClFeIII(HETP)] has been isolated and characterized by single-crystal X-ray diffraction. PMID- 14558795 TI - Aqueous self-assembly of unsymmetric Peptide bolaamphiphiles into nanofibers with hydrophilic cores and surfaces. AB - Unsymmetric peptide bolaamphiphiles that incorporate (l-glutamyl)3glycine at one terminus and either tetraethylene glycol or aspartic acid at the other were found to form hydrogels at low wt %, presumably by self-assembling into nanofibers presenting (l-glutamyl)3glycine at their surfaces and burying the second headgroup at their cores. Transmission electron microscopy measurements on 1 wt % gels negatively stained with phosphotungstic acid and positively stained with uranyl acetate show one-dimensional objects with diameters of 5 nm and lengths in excess of 1 mum. Circular dichroism and solid-state FTIR spectra indicate the adoption of beta-sheet structure within the nanofibers. PMID- 14558796 TI - Assembly of a supramolecular cube, [(Cp*WS3Cu3)8 Cl8(CN)12Li4] from a preformed incomplete cubane-like compound [PPh4][Cp*WS3(CuCN)3]. AB - A supramolecular cube has been formed by linking WS3Cu3 clusters with bidentate ligands. Eight WS3Cu3 clusters, which have an incomplete cubane-like structure, serve as the three connecting nodes of the cube, while 12 bridging cyanide anions coordinated to copper centers lie along the edges of the cube. Eight chloride anions and four lithium cations are located inside the cube. PMID- 14558797 TI - Tetrahedranyllithium: synthesis, characterization, and reactivity. AB - The first representative of stable tetrahedranyl anion, tris(trimethylsilyl)tetrahedranyllithium (3), has been synthesized by the reaction of tetrakis(trimethylsilyl)tetrahedrane (2) with methyllithium in tetrahydrofuran. The structural characterization of the tetrahedranyllithium has been achieved by X-ray crystallography, showing that the structure of 3.(TMEDA)1.5 represents a stretched tetrahedron. The endocyclic C(Li)-C(SiMe3) bond lengths range from 1.5408(15) to 1.5441(15) A (av 1.5425(15) A), and are longer than the endocyclic C(SiMe3)-C(SiMe3) bond lengths, which range from 1.4961(15) to 1.5009(15) A (av 1.4986(15) A). Methyl- and hydrogen-substituted tetrahedranes have also been prepared by the reaction of 3 with dimethyl sulfate and cyclopentadiene, respectively. PMID- 14558798 TI - Shell cross-linked cylinders of polyisoprene-b-ferrocenyldimethylsilane: formation of magnetic ceramic replicas and microfluidic channel alignment and patterning. AB - Stable organometallic cylindrical nanostructures have been prepared by shell cross-linking of PI320-b-PFS53 wormlike micelles in hexane through metal catalyzed hydrosilylation at 23 degrees C. The cross-linked structures permit the formation of cylindrical ceramic replicas containing size- and separation-tunable arrays of Fe nanoclusters. In addition, microfluidic alignment of the cross linked cylinders was possible. PMID- 14558799 TI - [(Th2F5)(NC7H5O4)2(H2O)][NO3]: an actinide-organic open framework. AB - We believe [(Th2F5)(NC7H5O4)2(H2O)][NO3] is the first three-dimensionally connected, actinide-organic framework solid. The structure is composed of thorium oxyfluoride chains, each of which connected to pyridinedicarboxylate groups to give a 3D cross-linked network with cavities containing nitrate anions. PMID- 14558800 TI - Ultrathin cross-linked nanoparticle membranes. AB - The fabrication of functional nanostructured materials for sensing, encapsulation and delivery requires practical approaches to self-assembly on multiple length scales and the synthesis of tough yet permeable structures. Here, the self assembly of functionalized, photoluminescent nanoparticles at liquid interfaces, followed by cross-linking of the associated ligands, affords robust membranes that maintain their integrity even when they are removed from the interface. These composite membranes, nanometers in thickness, are elastic yet permeable and have potential applications involving controlled permeability and diffusion. Cadmium selenide (CdSe) nanoparticles are used, since their inherent photoluminescence offers a direct way to probe the spatial organization of the particles. Functionalized ligands attached to the nanoparticles provide an effective means to stabilize the interfacial assembly by cross-linking. The concepts shown are adaptable to other type of nanoparticles, ligands, and solvent combinations. PMID- 14558802 TI - Stereoselective intramolecular [4 + 3] cycloadditions of nitrogen-stabilized chiral oxyallyl cations via epoxidation of N-tethered allenamides. AB - The first intramolecular [4 + 3] cycloaddition reaction using nitrogen-stabilized chiral oxyallyl cations that are tethered to furan or diene through the nitrogen atom is described here. Formation of these nitrogen-stabilized chiral oxyallyl cations is achieved by a chemoselective epoxidation of chiral allenamides via syringe pump addition of dimethyl dioxirane. The ensuing cycloaddition can be carried out with a range of different lengths for the tether, and high diastereoselectivities can be obtained when using chiral allenamides with shorter tethers. PMID- 14558801 TI - A new copper acetate-bis(oxazoline)-catalyzed, enantioselective Henry reaction. AB - A highly enantioselective, nitroaldol reaction catalyzed by a chiral Cu(II) bis(oxazoline) complex has been developed. The reaction scope includes both aromatic and aliphatic aldehydes (15 examples) affording products in good yields and enantioselectivities (87-94% ee). An X-ray structure of the catalyst has been provided along with a rationalization of the sense of asymmetric induction. PMID- 14558803 TI - Three-dimensional self-assembly of metallic rods with submicron diameters using magnetic interactions. AB - Metallic rods with submicron diameters that contain disklike ferromagnetic sections self-assemble into highly stable, hexagonally close-packed arrays of rods. The rods were fabricated by electrodeposition in porous alumina membranes and comprised alternating sections of gold and nickel. The thicknesses of the ferromagnetic nickel sections were approximately one-half the diameter of the rods (400 nm); this geometry orients the "easy" axis of magnetization perpendicular to the long axis of the rod. After magnetization of the rods with a rare-earth magnet, followed by sonication of the suspension, the rods spontaneously assembled into three-dimensional (3D) bundles that, on average, contained 15-30 rods. A macroscopic model of the rods suggests that the most stable orientation of the magnetic dipoles for rods in a defect-free, hexagonally close-packed arrangement is in concentric rings with the dipoles oriented head-to tail. This configuration minimizes the energy of the bundle and does not generate a net dipole for the structure. This work provides a simple demonstration that magnetic interactions between ferromagnetic objects can direct and stabilize the formation of ordered, 3D structures by self-assembly. PMID- 14558804 TI - New insights into the reaction of zinc alkyls with dioxygen. AB - Studies on the reaction of zinc alkyls with O2 are reported which demonstrate that the selective oxygenation of organozinc compound is viable. The reaction of [EtZn(azol)]n (azol = deprotonated 1-aziridineethanol) with an excess of dry O2 in toluene affords the zinc ethylperoxide [EtOOZn(azol)]2[EtZn(azol)]2, while the analogous reaction between Me2Zn and O2 results in the isolation of the Me6Zn7(OMe)8 cluster in high yield. PMID- 14558805 TI - Cine-substitution in the Stille coupling: evidence for the carbenoid reactivity of sp3-gem-organodimetallic iodopalladio-trialkylstannylalkane intermediates. AB - Two complementary routes to sp3-gem-organodimetallic iodopalladio trialkylstannanylalkanes are presented. Such intermediates have been proposed as Pd-carbenoid precursors in the Busacca-Farina cine-substitution mechanism in the Stille coupling. The decomposition of iodomethyltrialkylstannanes by Pd(0) catalysts was monitored by 1H, 2D, and 119Sn NMR. The formation of ethylene, trace formaldehyde, and iodotrialkylstannanes was detected. When the reaction was carried out in the presence of norbornene, the corresponding cyclopropane was produced in good yield. These observations are consistent with the intermediacy of a Pd-carbenoid species. sp3-gem-Organodimetallic iodopalladio trialkylstannanylalkane complexes were also prepared under stoichiometric conditions via transmetalation from tin to Pd(II). Me3SnCH2Sn(CH2CH2CH2)3 reacted with [(D-t-BPF)PdI]+I-, yielding the (D-t-BPF)Pd(II)ICH2SnMe3 complex that dimerized to form ethylene and cyclopropanated norbornene. The carbenoid reactivity of iodopalladio-trialkylstannanylalkanes complexes validates the Busacca-Farina mechanism of the cine-substitution in the Stille coupling. PMID- 14558806 TI - Mechanistic studies of the addition of carbonyl compounds to tetramesityldigermene. AB - The mechanism of the addition of carbonyl compounds to digermenes was investigated by examining the reaction between tetramesityldigermene and two mechanistic probes: trans-2-phenylcyclopropane carbaldehyde and trans,trans-2 methoxy-3-phenylcyclopropane carbaldehyde. In each reaction, two diastereomers of the formal [2+2] cycloadduct between the aldehyde and the digermene were obtained. These findings, together with the results of a recent theoretical study, indicate that the addition of carbonyl compounds to digermenes proceeds via a mechanism involving a zwitterionic intermediate. PMID- 14558807 TI - Sub-100 nm confinement of magnetic nanoparticles using localized magnetic field gradients. AB - Ferromagnetic rods containing thin sections of diamagnetic metal create intense magnetic field gradients that attract and confine magnetic nanoparticles to regions of space as small as 20 nm. The rods (80 nm diameter) comprised alternating sections of CoNi ( approximately 350 nm) and Au (20-160 nm) formed by electrodeposition into porous polycarbonate membranes. Upon magnetizing the rods, large magnetic gradients (106-107 T/m) form at the boundaries between ferromagnetic and diamagnetic sections. These gradients attract and confine magnetic nanoparticles to attoliter volumes of space surrounding the rod. This method provides a new tool for generating intense, highly localized magnetic field gradients, by design, and confining magnetic nanoparticles in these gradients. PMID- 14558808 TI - An experimental look into subelectron charge flow. AB - The prediction and measurement of charge distribution among interacting chemical entities in complex environments is a major challenge for modern chemistry. It encompasses information concerning fundamental quantities such as the electronic chemical potential and hardness of molecular fragments as well as their interactions with the surroundings. Although a wealth of theoretical work has been accumulated from the days of Pauling to the present, a specific molecular model system that allows quantitative and direct measurement of these properties has not yet been reported. Because atomic charges are not quantum mechanical observables, they cannot be derived from first principles, but rather they rely on the availability of high-precision experimental data and the interpretation of related experimental observables. Here, we demonstrate, for the first time, that a fragmental charge flow between a chelated metal center and reversibly bound molecules can be accurately monitored experimentally. PMID- 14558809 TI - Polyketide chain length control by chain length factor. AB - Bacterial aromatic polyketides are pharmacologically important natural products. A critical parameter that dictates product structure is the carbon chain length of the polyketide backbone. Systematic manipulation of polyketide chain length represents a major unmet challenge in natural product biosynthesis. Polyketide chain elongation is catalyzed by a heterodimeric ketosynthase. In contrast to homodimeric ketosynthases found in fatty acid synthases, the active site cysteine is absent from the one subunit of this heterodimer. The precise role of this catalytically silent subunit has been debated over the past decade. We demonstrate here that this subunit is the primary determinant of polyketide chain length, thereby validating its designation as chain length factor. Using structure-based mutagenesis, we identified key residues in the chain length factor that could be manipulated to convert an octaketide synthase into a decaketide synthase and vice versa. These results should lead to novel strategies for the engineered biosynthesis of hitherto unidentified polyketide scaffolds. PMID- 14558810 TI - Ab initio excited-state dynamics of the photoactive yellow protein chromophore. AB - The photoisomerization mechanism of the neutral form of the photoactive yellow protein (PYP) chromophore is investigated using ab initio quantum chemistry and first-principles nonadiabatic molecular dynamics (ab initio multiple spawning or AIMS). We identify the nature of the two lowest-lying excited states, characterize the short-time behavior of molecules excited directly to S2, and explain the origin of the experimentally observed wavelength-dependent photoisomerization quantum yield. PMID- 14558811 TI - Naphtho[2,3-a]pyrene forms chiral domains on Au111. AB - Chiral domains have been prepared by evaporation of a two-dimensionally chiral molecule, naphtho[2,3-a]pyrene (NP), onto the hexagonal Au(111) surface in an ultrahigh vacuum environment. High-resolution UHV scanning tunneling microscopy (STM) showed that NP formed chiral domains consisting of only one 2D enantiomer rather than racemic two-dimensional unit cells. A structural model is proposed that agrees with the STM observations. Chiral pockets between adsorbed molecules may be useful for binding a specific enantiomer of a 3D chiral molecule. PMID- 14558812 TI - Nonspecificity induces chiral specificity in the folding transition of giant DNA. AB - The all-or-none type DNA folding transition from a coil to globule can be differentiated by the chirality of the triamines. The fluorescent microscope observation on single DNA molecules makes it clear that the tripeptides obtained from naturally occurring basic amino acids (l-lysine or l-arginine) can compact DNA molecules at concentrations lower than those from d-isomers. Nanometer-sized beads are found in the AFM images on the folded DNA molecule. PMID- 14558813 TI - Bis(carbene)pyridine complexes of Cr(III): exceptionally active catalysts for the oligomerization of ethylene. AB - Pincer-heterocyclic carbene complexes of Cr(III), of the form [2,6-(1 alkylimidazol-2-ylidene)pyridine]CrCl3, have been prepared and evaluated as catalysts for the oligomerization of ethylene to alpha-olefins. In combination with methylaluminoxane cocatalyst, exceptionally high activities are obtained, ranging up to ca. 40 000 g mmol-1 bar-1 h-1. PMID- 14558815 TI - Synthesis of furanophane derivatives through [8+2]-cycloaddition of dienylisobenzofurans and alkynes. AB - The coupling of various dienylisobenzofurans with dimethyl acetylenedicarboxylate (DMAD) has been examined. In most cases, this reaction proceeds via [8+2] cycloaddition to afford furan-bridged decatetraene ring systems. The major competing reaction pathway is [4+2]-cycloaddition between DMAD and the isobenzofuran nucleus. Isobenzofuran intermediates were generated using either a chromium carbene-based method or an acid-catalyzed method. PMID- 14558814 TI - Hydrophobically directed selective reduction of ketones. AB - Three hydrophobic borohydrides carrying phenyl, pentafluorophenyl, and beta naphthyl groups were used to reduce ketones in water and in methanol. With ketones carrying phenyl, naphthyl, or biphenyl substituents, there was preferential reduction in methanol of competing acetyl groups, either intermolecular or intramolecular, but preferential reduction of the aryl ketones in water. In the most extreme case, there was a 40-fold selectivity reversal. Lithium borohydride showed no such change in selectivity and favored acetyl reduction in both solvents. Salt and cosolvent effects indicate that hydrophobic packing is involved in the reaction of hydrophobic reagents with the aryl ketones. Some special interaction of the pentafluorophenyl group with aryl rings was also detected. PMID- 14558816 TI - Biomolecular stress-sensitive gauges: surface-mediated immobilization of mechanosensitive membrane protein. AB - We report the observation of structural reorganizations associated with unique, stress-assisted gating of mechanosensitive (MscL) membrane protein on a silicon surface modified with alkyl-terminated monolayers. We observed that the shape of MscL membrane proteins changed dramatically depending upon the packing density of alkyl tails and the surface tension of the supporting organic layer. High resolution atomic force microscopy confirmed a transition from an elongated, prolate shape of MscL molecules within a monolayer with low surface tension to a flattened, oblate shape with a wide central opening within a monolayer with high surface tension. These observations are consistent with the conformation reorganizations associated with the two-stage, "iris"-like expansion proposed for the gating of the MscL molecules. PMID- 14558817 TI - Asymmetric dimers can be formed by dewetting half-shells of gold deposited on the surfaces of spherical oxide colloids. AB - Asymmetric dimers consisting of gold microcrystals and spherical silica colloids have been fabricated by depositing thin films of gold onto the spherical colloids to form half-shells, followed by annealing at elevated temperatures. The capability and feasibility of this procedure have been demonstrated with silica and titania beads of 0.2-2 mum in diameter and gamma-Fe2O3/polystyrene@SiO2 core shell particles 0.5 mum in size. The dimensions of gold microcrystals could be conveniently varied in the range of 100-650 nm by controlling the thickness of gold films and/or the diameter of the spherical colloids. This method provides another route to asymmetric dimers made of colloidal particles that could be different in size, chemical composition, surface functionality, density or sign of surface charge, bulk property, or a combination of these properties. PMID- 14558818 TI - Covalent adlayer growth on a diamond thin film surface. AB - The surface of boron-doped diamond thin films can be modified by exposure to a strong oxidizing agent, resulting in the formation of -OH and =O terminated diamond. The -OH groups are reacted with an acid chloride to produce a covalently bound, modified diamond thin film surface. The demonstration of these reactions allows for the facile modification of diamond surfaces using techniques well established for oxide surfaces. Characterization of the covalently bound species shows submonolayer coverage, and time-resolved fluorescence measurements are reflective of the highly featured nature of the diamond film. PMID- 14558819 TI - Stepwise evolution of the structure and electronic properties of DNA. AB - The optical properties of a family of duplex DNA conjugates possessing from 1 to 11 A:T base pairs and covalently attached stilbene chromophores at both ends have been investigated. As few as from two to four base bairs are needed to establish base-pair absorption and circular dichroism spectra similar to those of longer oligomers. The stilbene chromophores exhibit exciton-coupled circular dichroism, which can be observed even with 11 intervening base pairs, a distance of ca. 40 A. The dependence of the sign and amplitude of these spectra reveals the ability of B-DNA to serve as a helical ruler. PMID- 14558820 TI - Dual catalytic apparatus of the thiamin diphosphate coenzyme: acid-base via the 1',4'-iminopyrimidine tautomer along with its electrophilic role. AB - It was recently reported (Jordan, F.; Zhang, Z.; Sergienko, E. A. Bioorg. Chem. 2002, 30, 188-198) that addition to the E477Q active-center variant of yeast pyruvate decarboxylase of (a) pyruvate on a rapid-scan UV stopped-flow, or (b) acetaldehyde or benzoylformate on a circular dichroism (CD) instrument, generates a new band with lambda(max) near 300-310 nm. A chemical model demonstrated that the wavelength is appropriate to the 1',4'-iminopyrimidine tautomer of the 4' aminopyrimidine ring in thiamin diphosphate. Herein, we report the formation of a new positive CD band centered at 305 nm when the Escherichia colipyruvate dehydrogenase complex first E1 subunit and its variants are exposed to phosphonolactylthiamin diphosphate, a stable analogue of the covalent adduct formed between the substrate pyruvate and the C2 atom of thiamin diphosphate. The behavior of this CD band, whether it suggests saturation of the enzyme by phosphonolactylthiamin diphosphate, or its very existence (the band is not seen with the E571A E1 variant, where E571 is hydrogen bonded to the N1' atom of the 4'-aminopyrimidine ring), as well as its position are consistent with its assignment to the 1',4'-imino thiamin diphosphate tautomer on the enzyme, chiral by virtue of its fixed V conformation. The mechanism of binding of phosphonolactylthiamin diphosphate closely resembles that of thiamin diphosphate itself. PMID- 14558821 TI - NMR and quantum chemical study on the OH...pi and CH...O interactions between trehalose and unsaturated fatty acids: implication for the mechanism of antioxidant function of trehalose. AB - Trehalose is a disaccharide that attracts much attention as a stress protectant. In this study, we investigated the mechanism of the antioxidant function of trehalose. The spin-lattice relaxation times (T(1)) of (1)H and (13)C NMR spectra were measured to investigate the interaction between trehalose and unsaturated fatty acid (UFA). We selected several kinds of UFA that differ in the number of double bonds and in their configurations (cis or trans). Several other disaccharides (sucrose, maltose, neotrehalose, maltitol, and sorbitol) were also analyzed by NMR. The T(1) values for the (1)H and (13)C signals assigned to the olefin double bonds in UFA decrease with increasing concentration of trehalose and the changes reaches plateaus at integer ratios of trehalose to UFA. The characteristic T(1) change is observed only for the combination of trehalose and UFA with cis double bond(s). On the other hand, from the (13)C-T(1) measurements for trehalose, the T(1) values of the C-3 (C-3') and C-6' (C-6) are found to change remarkably by addition of UFA. (1)H[bond](1)H NOESY measurements provide direct evidence for complexation of trehalose with linoleic acid. These results indicate that one trehalose molecule stoichiometrically interacts with one cis olefin double bond of UFA. Computer modeling study indicates that trehalose forms a stable complex with an olefin double bond through OH...pi and CH...O types of hydrogen bonding. Furthermore, a significant increase in the activation energy is found for hydrogen abstraction reaction from the methylene group located between the double bonds that are both interacting with the trehalose molecules. Therefore, trehalose has a significant depression effect on the oxidation of UFA through the weak interaction with the double bond(s). This is the first study to elucidate the antioxidant function of trehalose. PMID- 14558822 TI - Synthesis and biological activity of photoactivatable N-ras peptides and proteins. AB - A modular strategy for the assembly of farnesylated N-Ras heptapeptides carrying a photoactivatable benzophenone (BP) group within the lipid residue is described. This strategy is based on the fragment condensation of a N-terminal hexapeptide synthesized on the solid support with a cysteine methyl ester which is modified with different farnesyl analogues, incorporating the photophor. At the N-terminus of the peptides different functional groups can be attached, e.g., biotin for product enrichment and detection after photoactivation or a maleimido (MIC) linker, allowing for the coupling to proteins carrying a C-terminal free cysteine. Using this strategy, 24 peptides were synthesized, incorporating farnesyl analogues with four different chain lengths. Two of these photoactivatable conjugates were ligated to oncogenic human N-RasG12V Delta 181. A cellular transformation assay revealed that the semisynthetic proteins retain their biological activity despite the photolabel. The first photolabeling experiments with a geranyl-BP-labeled N-Ras construct and the farnesyl-sensitive guanine nucleotide exchange factor hSos1 indicate that this photoaffinity labeling system can be particularly useful for studying protein-protein interactions, e.g., the participation of the farnesyl group in Ras signaling, which is still discussed with controversy. PMID- 14558823 TI - 5-enolpyruvylshikimate 3-phosphate synthase: chemical synthesis of the tetrahedral intermediate and assignment of the stereochemical course of the enzymatic reaction. AB - A chemical synthesis of both diastereomers of the tetrahedral intermediate involved in 5-enolpyruvylshikimate 3-phosphate synthase (EPSPS) catalysis has been accomplished. Combination of methyl dibromopyruvate with a protected shikimic acid derivative, phosphorylation, and lactonization afforded the intermediates (S)-15 and (R)-15, whose configurations were assigned by NMR. After introduction of the 3-phosphate group and deprotection, photoinitiated radical debromination of the dibromo analogues (S)-5 and (R)-5 was accomplished with tributyltin hydride in mixed aqueous solvents in the presence of surfactant to give the pyruvate ketal phosphates (R)-TI and (S)-TI, respectively. These compounds are stable at high pH, but decompose at pH 7 with a half-life of ca. 10 min. (R)-TI proved to be inert to EPSPS, while (S)-TI was converted by the enzyme to a mixture of 5-enolpyruvylshikimate 3-phosphate, shikimate 3-phosphate, and phosphoenolpyruvate. The demonstration that the enzymatic intermediate possesses the S-configuration at the ketal center confirms the mechanism as an anti addition followed by a syn elimination. Furthermore, it appears that the syn stereochemistry of the second step requires the phosphate leaving group to serve as the base in catalyzing its own elimination. PMID- 14558824 TI - Balancing kinetic and thermodynamic control: the mechanism of carbocation cyclization by squalene cyclase. AB - Molecular dynamics simulations with a combined quantum mechanical and molecular mechanical (QM/MM) potential have been carried out to investigate the squalene-to hopene carbocation cyclization mechanism in squalene-hopene cyclase (SHC). The present study is based on free energy simulations by constructing the free energy surface for the cyclization steps along the reaction pathway. The picture that emerges for the carbocation cyclization cascade is a delicate balance of thermodynamic and kinetic control that ultimately favors the formation of the final hopanoids carbon skeleton. A key finding is that the five- to six-membered ring expansion process is not a viable reaction pathway for either C- or D-ring formation in the cyclization reaction. The only significant intermediate is the A/B-bicyclic cyclohexyl cation (III), from which two asynchronous concerted reaction pathways lead to, respectively, the 6,6,6,5-tetracyclic carbon skeleton and the 6,6,6,6,5-pentacyclic hopanoids. Experimentally, these two products are observed to have 1% and 99% yields, respectively, in the wild-type enzyme. We conclude that the product distribution in the wild-type enzyme is dictated by kinetic control of these two reaction pathways. PMID- 14558825 TI - The Vroman effect: a molecular level description of fibrinogen displacement. AB - The molecular level details of the displacement of surface adsorbed fibrinogen from silica substrates were studied by atomic force microscopy, immunochemical assays, fluorescence microscopy, and vibrational sum frequency spectroscopy. The results showed that human plasma fibrinogen (HPF) can be readily displaced from the interface by other plasma proteins near neutral pH because the positively charged alpha C domains on HPF sit between the rest of the macromolecule and the underlying surface. The alpha C domains make weak electrostatic contact with the substrate, which is manifest by a high degree of alignment of Lys and Arg residues. Upon cycling through acidic pH, however, the alpha C domains are irreversibly removed from this position and the rest of the macromolecule is free to engage in stronger hydrogen bonding, van der Waals, and hydrophobic interactions with the surface. This results in a 170-fold decrease in the rate at which HPF can be displaced from the interface by other proteins in human plasma. PMID- 14558826 TI - Atomic resolution density maps reveal secondary structure dependent differences in electronic distribution. AB - The X-ray crystal structure of the flavoenzyme cholesterol oxidase, SCOA (Streptomyces sp.SA-COO) has been determined to 0.95 A resolution. The large size (55kDa) and the high-resolution diffraction of this protein provides a unique opportunity to observe detailed electronic effects within a protein environment and to obtain a larger sampling for which to analyze these electronic and structural differences. It is well-known through spectroscopic methods that peptide carbonyl groups are polarized in alpha-helices. This electronic characteristic is evident in the sub-Angstrom electron density of SCOA. Our analysis indicates an increased tendency for the electron density of the main chain carbonyl groups within alpha-helices to be polarized toward the oxygen atoms. In contrast, the carbonyl groups in beta-sheet structures tend to exhibit a greater charge density between the carbon and oxygen atoms. Interestingly, the electronic differences observed at the carbonyl groups do not appear to be correlated to the bond distance of the peptide bond or the peptide planarity. This study gives important insight into the electronic effects of alpha-helix dipoles in enzymes and provides experimentally based observations that have not been previously characterized in protein structure. PMID- 14558827 TI - Independent generation of 5-(2'-deoxycytidinyl)methyl radical and the formation of a novel cross-link lesion between 5-methylcytosine and guanine. AB - Reactive oxygen species (ROS) can damage DNA. Although a number of single nucleobase lesions induced by ROS have been structurally characterized, only a few intrastrand cross-link lesions have been identified and characterized, and all of them involve adjacent thymine and guanine or adenine. In mammalian cells, the cytosines at CpG sites are methylated. On the basis of the similar reactivity of 5-methylcytosine and thymine toward hydroxyl radical and the similar orientation of adjacent thymine guanine (TG) and 5-methylcytosine guanine (mCG) in B-DNA, we predict that the cross-link lesion, which was identified in TG and has a covalent bond formed between the 5-methyl carbon atom of T and the C8 carbon atom of G, should also form at mCG site. Here, we report for the first time the independent generation of 5-(2'-deoxycytidinyl)methyl radical, and our results demonstrate that this radical can give rise to the predicted novel intrastrand cross-link lesion in dinucleoside monophosphates d(mCG) and d(GmC). Furthermore, we show that the cross-link lesion can also form in d(mCG) from gamma irradiation under anaerobic conditions. PMID- 14558828 TI - Active oxygen species generated from photoexcited fullerene (C60) as potential medicines: O2-* versus 1O2. AB - To characterize fullerenes (C(60) and C(70)) as photosensitizers in biological systems, the generation of active oxygen species, through energy transfer (singlet oxygen (1)O(2)) and electron transfer (reduced active oxygen radicals such as superoxide anion radical O(2)(-)* and hydroxyl radical *OH), was studied by a combination of methods, including biochemical (DNA-cleavage assay in the presence of various scavengers of active oxygen species), physicochemical (EPR radical trapping and near-infrared spectrometry), and chemical methods (nitro blue tetrazolium (NBT) method). Whereas (1)O(2) was generated effectively by photoexcited C(60) in nonpolar solvents such as benzene and benzonitrile, we found that O(2)(-)* and *OH were produced instead of (1)O(2) in polar solvents such as water, especially in the presence of a physiological concentration of reductants including NADH. The above results, together with those of a DNA cleavage assay in the presence of various scavengers of specific active oxygen species, indicate that the active oxygen species primarily responsible for photoinduced DNA cleavage by C(60) under physiological conditions are reduced species such as O(2)(-)* and *OH. PMID- 14558829 TI - Side-chain dynamics of an alpha-helical polypeptide monitored by fluorescence. AB - The "blob" model, developed to analyze the fluorescence decays of polymers randomly labeled with pyrene, has been applied to a series of pyrene-labeled poly(glutamic acid)s (PyPGA) in DMF and carbonated buffer solutions at pH 9. Poly(glutamic acid) (PGA) exists in the ionized form in the buffer solutions as poly(sodium glutamate) (PGNa). PGA adopts an alpha-helical conformation in DMF, whereas in aqueous solution PGNa is a random coil. Fluorescence, UV-vis absorption, and circular dichroism measurements indicate that in our studies pyrene pendants attached themselves along PGA in a clustered manner. Simulations were carried out to establish that the geometry of the PGA alpha-helix induces the high level of pyrene clustering. Since the level of pyrene clustering decreased with lower pyrene content, information about naked PGA was retrieved by extrapolating the trends obtained by fluorescence to zero pyrene content. Analysis of the fluorescence decays demonstrated that during its lifetime an excited pyrene probes a 32 amino acid section of the PGA alpha-helix. This result was supported by molecular mechanics optimizations. This study establishes that the blob model, originally used to monitor the encounters between pyrenes attached randomly onto a polymer adopting a random coil conformation, can also be applied to study the dynamics of the side chains of structured proteins. Since the blob model helps in monitoring the encounters between amino acids in the initial state (i.e., random coil) and in the final state (i.e., structured protein) of the folding pathway of a protein, it could be applicable to the study of protein folding. PMID- 14558830 TI - DFT calculations on the effects of para substituents on the energy differences between singlet and triplet states of 2,2-difluoro-1,3-diphenylcyclopentane-1,3 diyls. AB - UB3LYP/6-31g* calculations have been performed on a series of para-substituted 2,2-difluoro-1,3-diphenylcyclopentane-1,3-diyls (4). The singlet is computed to be the ground state for each of the diradicals, regardless of the nature of the para substituents, which range from strongly pi-electron-donating (amino) to strongly pi-electron-withdrawing (nitro). In the symmetrically para-disubstituted diradicals, the size of the singlet-triplet energy gap (Delta E(ST)) increases with the pi-electron-donating ability of the substituents, but in the unsymmetrically substituted diradicals, large values of Delta E(ST) are calculated even when one of the substituents is a pi electron acceptor. The origins of the competitive and cooperative substituent effects, predicted for diradical 4, are discussed in light of the calculated effects of the same substituents on the singlet and triplet states of diradical 6, which lacks the geminal fluorines at C-2 that are present in 4. PMID- 14558831 TI - Biphenalenylidene: the forgotten bistricyclic aromatic ene. A theoretical study. AB - The Lawesson reagent and P(2)S(5) mediated reductive coupling of phenalenone (6) gives LPAH peropyrene (5) in 47% and 54% yields. The mechanism of the reaction involves the formation of phenalenethione (10), Z- and E-1,1'-biphenalenylidene (3), and 9 as intermediates. The electrocyclization reaction of Z-3 to 9, followed by aromatization, gives 5. The results of an ab initio and DFT study of 3 and 2,2'-biphenalenylidene (12) are reported. E-3 and Z-3 have a diradical character with twist angles of 44.8 degrees and 57.8 degrees (at UB3LYP/6 311G**). Delta E(++)(Tot) = 10.2 kJ/mol and Delta G(++)(298) = 10.6 kJ/mol for E 3 <==> Z-3 diastereomerization. These unusually low energy barriers are due to the ground-state diradical destabilization and the aromatic stabilization of the transition state TS-3. Triplet Z-3 is higher in energy than singlets E-3 and Z-3 by 10.4 and 3.1 kJ/mol. In the concealed non-Kekule 12, singlet 12 is more stable than the triplet by 1.3 kJ/mol. Singlet 12 is more stable than singlet E-3 by 2.0 kJ/mol, and orthogonal singlet TS-12 is lower in energy than singlet TS-3 by 6.0 kJ/mol. The energy barriers for the hexatriene-cyclohexadiene electrocyclization Z-3 --> 9 are Delta E(++)(Tot) = 94.8 and Delta G(++)(298) = 98.3 kJ/mol (at (U)B3LYP/6-31G). The reaction occurs thermally in a conrotatory mode. PMID- 14558832 TI - A second-generation synthesis of the C1-C28 portion of the altohyrtins (spongistatins). AB - A practical second-generation synthesis of an advanced intermediate in our total synthesis of altohyrtin C (spongistatin 2) has been developed. A new approach to the C1-C15 (AB) portion features a vinyllithium addition to an aldehyde followed by a palladium-catalyzed allylic reduction to install the troublesome C13-C15 segment. Our general approach to the C16-C28 (CD) spiroketal has been retained, but some improvements have been made. Most notably, the kinetically controlled CD spiroketalization reaction now proceeds in high yield with excellent diastereoselection. This new strategy uses the anti-aldol coupling used in our first-generation synthesis to join AB and CD fragments. A total of 9.6 g of intermediate 57 has been produced using this improved route. PMID- 14558833 TI - Multigram synthesis of the C29-C51 subunit and completion of the total synthesis of altohyrtin C (spongistatin 2). AB - A multigram synthesis of the C29-C51 subunit of altohyrtin C (spongistatin 2) has been accomplished. Union of this intermediate with the C1-C28 fragment and further elaboration furnished the natural product. Completion of the C29-C51 subunit began with the aldol coupling of the boron enolate derived from methyl ketone 8 and aldehyde 9. Acid-catalyzed deprotection/cyclization of the resulting diastereomeric mixture of addition products was conducted in a single operation to afford the E-ring of altohyrtin C. The diastereomer obtained through cyclization of the unwanted aldol product was subjected to an oxidation/reduction sequence to rectify the C35 stereocenter. The C45-C48 segment of the eventual triene side chain was introduced by addition of a functionalized Grignard reagent derived from (R)-glycidol to a C44 aldehyde. Palladium-mediated deoxygenation of the resulting allylic alcohol was followed by adjustment of protecting groups to provide reactivity suitable for the later stages of the synthesis. The diene functionality comprising the remainder of the C44-C51 side chain was constructed by addition of an allylzinc reagent to the unmasked C48 aldehyde and subsequent dehydration of the resulting alcohol. Completion of the synthesis of the C29-C51 subunit was achieved through conversion of the protected C29 alcohol into a primary iodide. The synthesis of the C29-C51 iodide required 44 steps with a longest linear sequence of 33 steps. From commercially available tri-O-acetyl-d glucal, the overall yield was 6.8%, and 2 g of the iodide was prepared. The C29 C51 primary iodide was amenable to phosphonium salt formation, and the ensuing Wittig coupling with a C1-C28 intermediate provided a fully functionalized, protected seco-acid. Selective deprotection of the required silicon groups afforded an intermediate appropriate for macrolactonization, and, finally, global deprotection furnished altohyrtin C (spongistatin 2). This synthetic approach required 113 steps with a longest linear sequence of 37 steps starting from either tri-O-acetyl-d-glucal or (S)-malic acid. PMID- 14558834 TI - Selective oxygenation of ring-substituted toluenes with electron-donating and withdrawing substituents by molecular oxygen via photoinduced electron transfer. AB - A ring-substituted toluene with an electron-withdrawing substituent, p tolunitrile, is oxygenated by molecular oxygen to yield the corresponding aldehyde with tetrafluoro-p-dicyanobenzene as a photocatalyst under photoirradiation with an Hg lamp (lambda > 300 nm). The oxygenation of a ring substituted toluene with an electron-donating substituent, p-xylene, by molecular oxygen is also achieved with 10-methyl-9-phenylacridinium ion as a photocatalyst under visible light irradiation, yielding p-tolualdehyde exclusively as the final oxygenated product. Both the oxygenation reactions are initiated by photoinduced electron transfer from the ring-substituted toluene to the singlet excited state of the photocatalyst. The reason for the high selectivity in the photocatalytic oxygenation of various toluene derivatives by molecular oxygen is discussed on the basis of the photoinduced electron transfer mechanism that does not involve the autoxidation process (radical chain reactions). The reactive intermediates in the photocatalytic cycle are successfully detected as the transient absorption spectra and the electron spin resonance spectra. PMID- 14558836 TI - Photoreactions of 3-diazo-3H-benzofuran-2-one; dimerization and hydrolysis of its primary photoproduct, a quinonoid cumulenone: a study by time-resolved optical and infrared spectroscopy. AB - Light-induced deazotization of 3-diazo-3H-benzofuran-2-one (1) in solution is accompanied by facile (CO)-O bond cleavage yielding 6-(oxoethenylidene)-2,4 cyclohexadien-1-one (3), which appears with a rise time of 28 ps. The expected Wolff-rearrangement product, 7-oxabicyclo[4.2.0]octa-1,3,5-trien-8 ylidenemethanone (4), is not formed. The efficient light-induced formation of the quinonoid cumulenone 3 opens the way to determine the reactivity of a cumulenone in solution. The reaction kinetics of 3 were monitored by nanosecond flash photolysis with optical (lambda(max) approximately 460 nm) as well as Raman (1526 cm(-1)) and IR detection (2050 cm(-)(1)). Remarkably, the reactivity of 3 is that expected from its valence isomer, the cyclic carbene 3H-benzofuran-2-one-3 ylidene, 2. In aqueous solution, acid-catalyzed addition of water forms the lactone 3-hydroxy-3H-benzofuran-2-one (5). The reaction is initiated by protonation of the cumulenone on its beta-carbon atom. In hexane, cumulenone 3 dimerizes to isoxindigo ((E)-[3,3']bibenzofuranylidene-2,2'-dione, 7), coumestan (6H-benzofuro[3,2-c][1]benzopyran-6-one, 8), and a small amount of dibenzonaphthyrone ([1]benzopyrano[4,3-][1]benzopyran-5,11-dione, 9) at a nearly diffusion-controlled rate. Ab initio calculations (G3) are consistent with the observed data. Carbene 2 is predicted to have a singlet ground state, which undergoes very facile, strongly exothermic (irreversible) ring opening to the cumulenone 3. The calculated barrier to formation of 4 (Wolff-rearrangement) is prohibitive. DFT calculations indicate that protonation of 3 on the beta-carbon is accompanied by cyclization to the protonated carbene 2H(+), and that dimerization of 3 to 7 and 9 takes place in a single step with negligible activation energy. PMID- 14558837 TI - Stoichiometric compounds of magnesium dichloride with ethanol for the supported Ziegler-Natta catalysis: first recognition and multidimensional MAS NMR study. AB - Ethanol associates easily with MgCl(2) to form adducts of complex architecture, but until now available characterization methods have failed to identify the pure stoichiometric compounds and their structures. To remedy this, we set about applying homonuclear and heteronuclear 2D correlated solid-state NMR spectroscopy to identify the pure compounds and the ethanol-to-magnesium coordination pattern. High spinning speed and Lee-Goldburg sequences were able to reduce the hydrogen spin-diffusion and homonuclear coupling in the crystalline solid, thus achieving high resolution also in the hydrogen domain. On this basis, the pure adducts, of interest as catalyst supports for Ziegler-Natta polymerization, were isolated for the first time. Magnesium coordination sites with given numbers of ligands and their multiplicity in the crystal cells were determined in the new-found stoichiometric complexes. Variable temperature and 2D carbon-carbon exchange NMR, as well as relaxation times in the fast motion regime, revealed the disordering phenomena generated by ethanol dynamics in the crystal. Decoding the intriguing polymorphism of the precursors permits to trace the genealogy of tailored MgCl(2) titanate granules, active as highly productive catalysts for the stereospecific polymerization of olefins. PMID- 14558835 TI - Synthesis of alpha-amino acids via asymmetric phase transfer-catalyzed alkylation of achiral nickel(II) complexes of glycine-derived Schiff bases. AB - Achiral, diamagnetic Ni(II) complexes 1 and 3 have been synthesized from Ni(II) salts and the Schiff bases, generated from glycine and PBP (7) and PBA (11), respectively, in MeONa/MeOH solutions. The requisite carbonyl-derivatizing agents pyridine-2-carboxylic acid(2-benzoyl-phenyl)-amide 7 (PBP) and pyridine-2 carboxylic acid(2-formyl-phenyl)-amide 11 (PBA) were readily prepared from picolinic acid and o-aminobenzophenone or picolinic acid and methyl o anthranilate, respectively. The structure of 1 was established by X-ray crystallography. Complexes 1 and 3 were found to undergo C-alkylation with alkyl halides under PTC conditions in the presence of beta-naphthol or benzyltriethylammonium bromide as catalysts to give mono- and bis-alkylated products, respectively. Decomposition of the complexes with aqueous HCl under mild conditions gave the required amino acids, and PBP and PBA were recovered. Alkylation of 1 with highly reactive alkyl halides, carried out under the PTC conditions in the presence of 10% mol of (S)- or (R)-2-hydroxy-2'-amino-1,1' binaphthyl 31a (NOBIN) and/or its N-acyl derivatives and by (S)- or (R)-2-hydroxy 8'-amino-1,1'-binaphthyl 32a (iso-NOBIN) and its N-acyl derivatives, respectively, gave rise to alpha-amino acids with high enantioselectivities (90 98.5% ee) in good-to-excellent chemical yields at room temperature within several minutes. An unusually large positive nonlinear effect was observed in these reactions. The Michael addition of acrylic derivatives 37 to 1 was conducted under similar conditions with up to 96% ee. The (1)H NMR and IR spectra of a mixture of the sodium salt of NOBIN and 1 indicated formation of a complex between the two components. Implications of the association and self-association of NOBIN for the observed sense of asymmetric induction and nonlinear effects are discussed. PMID- 14558839 TI - Novel halogen and interhalogen adducts of nanoscale magnesium oxide. AB - The difference in the adsorption properties of magnesium oxide in two different physical forms--large micron-sized crystals (CM-MgO) and nanoscale crystals (AP MgO)--is described. The highly energetic surface of the nanoparticles is significantly more active than the bulk surface in adsorption of nonpolar halogens (Cl(2), Br(2), and I(2)) and dipole interhalogen molecules (ICl, IBr, and ICl(3)). The nanocrystalline and microcrystalline MgO adducts with halogens and interhalogens were characterized by thermogravimetric analysis (TGA), UV-vis and Raman spectroscopies, and transmission electron microscopy (TEM). The bonds of the adsorbates were considerably strained upon adsorption on the nanocrystalline MgO without a molecular dissociation. The results presented in the paper explain the enhanced reactivities of the nanoscale MgO, halogen adducts as halogenating agents of organic molecules and their noticeable biocidal activity. PMID- 14558838 TI - L-edge X-ray absorption spectroscopy of non-heme iron sites: experimental determination of differential orbital covalency. AB - X-ray absorption spectroscopy has been utilized to obtain the L-edge multiplet spectra for a series of non-heme ferric and ferrous complexes. Using these data, a methodology for determining the total covalency and the differential orbital covalency (DOC), that is, differences in covalency in the different symmetry sets of the d orbitals, has been developed. The integrated L-edge intensity is proportional to the number of one-electron transition pathways to the unoccupied molecular orbitals as well as to the covalency of the iron site, which reduces the total L-edge intensity and redistributes intensity, producing shake-up satellites. Furthermore, differential orbital covalency leads to differences in intensity for the different symmetry sets of orbitals and, thus, further modifies the experimental spectra. The ligand field multiplet model commonly used to simulate L-edge spectra does not adequately reproduce the spectral features, especially the charge transfer satellites. The inclusion of charge transfer states with differences in covalency gives excellent fits to the data and experimental estimates of the different contributions of charge transfer shake-up pathways to the t(2g) and e(g) symmetry orbitals. The resulting experimentally determined DOC is compared to values calculated from density functional theory and used to understand chemical trends in high- and low-spin ferrous and ferric complexes with different covalent environments. The utility of this method toward problems in bioinorganic chemistry is discussed. PMID- 14558840 TI - Magnesium tetraarylporphyrin tweezer: a CD-sensitive host for absolute configurational assignments of alpha-chiral carboxylic acids. AB - A protocol to determine the absolute configuration of alpha-chiral carboxylic acids based on a modified circular dichroic (CD) exciton chirality method has been developed. The protocol relies on a host-guest complexation mechanism: the chiral substrates are derivatized to give bifunctional amide conjugates ("guests") that form complexes with a dimeric magnesium porphyrin host, Mg-T (T stands for "tweezer") that acts as a "receptor". The two porphyrins in the complex adopt a preferred helicity dictated by the substituents at the chiral center in accordance with their steric sizes (assigned on the basis of conformational energy A-values) and, consequently, with the absolute configuration of the substrates under investigation. This chiroptical method, verified with a variety of chiral substrates, has been demonstrated to be reliable and generally applicable, including natural products with complex structures. Molecular modeling, NMR, and FTIR experiments of selected host-guest complexes revealed the mode of ligation of the substrates to the magnesium porphyrin species and led to clarification of the structure of the complex. When oxygen functionalities were directly attached to the chiral center, the signs of the CD couplets were opposite to those predicted on the basis of steric size. NMR and molecular modeling experiments indicated that this apparent inconsistency was due to conformational characteristics of the guest molecules. The stereochemical analysis is shown to be a sensitive technique, not only for the determination of absolute configurations of substrates but also for elucidation of their solution conformations. PMID- 14558841 TI - Size- and support-dependent electronic and catalytic properties of Au0/Au3+ nanoparticles synthesized from block copolymer micelles. AB - Supported Au nanoclusters synthesized from diblock copolymer micelles can be reliably prepared with well-controlled sizes and dispersions. For particles with diameters between approximately 1 and 6 nm, the particle size and the support were found to strongly influence the oxygen reactivity, the formation and stabilization of a metal-oxide, and the catalytic activity for electrooxidation of carbon monoxide. The smallest particles studied (1.5 nm) were the most active for electrooxidation of CO and had the largest fraction of oxygen associated with gold at the surface as measured by the Au(3+)/Au(0) X-ray photoemission intensities. Conducting and semiconducting substrates, ITO-coated glass and TiO(2), respectively, were associated with greater stabilization of Au(3+) oxide as compared to insulating, SiO(2), substrates. PMID- 14558842 TI - Toward binary nitrosyls: distinctly bent Fe[bond]N[bond]O linkages in base stabilized Fe(NO)3+ complexes. AB - Air- and moisture-sensitive Fe(NO)(3)(eta(1)-PF(6)) (1) may be conveniently prepared by treating Fe(NO)(3)Cl with 1 equiv of [Ag][PF(6)] in CH(2)Cl(2) or by reacting [NO][PF(6)] with excess iron filings in MeNO(2). Complex 1 is thermally sensitive both as a solid and in solutions, and is best handled below -20 degrees C. To isolate 1 reproducibly from MeNO(2) solutions it is necessary to remove all traces of propionitrile, which often occurs as an impurity in MeNO(2), because it reacts with Lewis-acidic 1 to form [Fe(NO)(3)(EtCN)][PF(6)] (2). If trace H(2)O is present during the synthesis of 1, some of the PF(6)(-) is converted to PO(2)F(2)(-), which is sufficiently Lewis basic that it captures two Fe(NO)(3)(+) fragments and forms [(ON)(3)Fe(mu-PO(2)F(2))Fe(NO)(3)][PF(6)] (3). Finally, Fe(NO)(3)(eta(1)-BF(4)) (4) can be obtained as a green microcrystalline powder by employing the same synthetic methodologies used to prepare 1. The new complexes 1 4 have been characterized by conventional spectroscopic methods, and the solid state molecular structures of 2, 3, and 4 and their parent compound, Fe(NO)(3)Cl, have been established by X-ray diffraction methods. The iron centers in the Fe(NO)(3) fragments in all these structures exhibit approximately tetrahedral coordination geometries, and the Fe-N-O linkages are distinctly nonlinear with bond angles in the range of 159 to 169 degrees. DFT calculations on Fe(NO)(3)(eta(1)-BF(4)) (4) confirm that its bent Fe-N-O links have an electronic origin and need not be attributed to other factors, such as packing forces in the crystal. Interestingly, the bending of the NO ligands results in an increase in the energy of the HOMO, relative to the linear case, but at the same time causes a decrease in energy of the HOMO-1 and the HOMO-2 molecular orbitals. This more than compensates for the higher energy of the HOMO, resulting in a lower energy structure. PMID- 14558843 TI - Fully localized mixed-valence oxidation products of molecules containing two linked dimolybdenum units: an effective structural criterion. AB - Two previously reported compounds [Mo(2)](CH(3)O)(2)M(CH(3)O)(2)[Mo(2)] (Cotton, F. A.; Liu, C. Y.; Murillo, C. A.; Wang, X. Inorg. Chem. 2003, 42, 4619), in which [Mo(2)] is an abbreviation for the quadruply bonded Mo(2)(4+) unit embraced by three (p-anisyl)NC(H)N(p-anisyl) anions and M = Zn (1) or Co (2), have been chemically oxidized. One-electron oxidation products [Mo(2)](CH(3)O)(2)M(CH(3)O)(2)[Mo(2)](PF(6)) (3, M = Zn; 4, M = Co) and the two electron oxidation product [Mo(2)](CH(3)O)(2)Zn(CH(3)O)(OH)[Mo(2)](PF(6))(2) (5) have been isolated and structurally characterized. As expected, oxidations occur at the dimolybdenum units. The mono-charged cations in 3 and 4 have asymmetric molecular structures with two distinct [Mo(2)] units. In each case, one of the [Mo(2)] units has a lengthened Mo-Mo bond distance of 2.151[1] A, as expected for one-electron oxidation, whereas the other remains unchanged at 2.115[1] A. These correspond to bond orders of 3.5 (sigma(2)pi(4)delta(1)) and 4.0 (sigma(2)pi(4)delta(2)), respectively. The crystallographic results thus show unambiguously that in the crystalline state, the mixed-valence compounds (3 and 4) are electronically localized and the unpaired electron is trapped on one [Mo(2)] unit. These results are supported by the EPR spectra. The doubly oxidized compound 5 has two equivalent [Mo(2)] units, both with a Mo-Mo bond distance of 2.149[1] A. EPR and magnetic susceptibility measurements for 5 indicate that there is no significant ferromagnetic or antiferromagnetic spin coupling and the species is valence-trapped. PMID- 14558844 TI - Highly emissive supramolecular oligo(p-phenylene vinylene) dendrimers. AB - pi-Conjugated oligo(p-phenylene vinylene) (OPV) guest molecules for interaction with dendritic hosts were synthesized and fully characterized by NMR spectroscopy, MALDI-TOF-MS, elemental analysis and optical measurements. The binding properties of the five different OPV guests to a N,N-bis[(3-adamantyl ureido) propyl] methylamine host have been investigated. The guests that contained an aryl urea glycine spacer were bound with the highest association constant. Subsequently, an adamantyl urea modified fifth generation poly(propylene imine) dendrimer was synthesized as a multivalent host which contains 32 N,N-bis[(3-adamantyl ureido) propyl] amine binding sites. Size exclusion chromatography showed that 32 of the OPV guests strongly bind to the fifth generation adamantyl functionalized dendritic host. In the case of the supramolecular dendritic host/guest system smooth homogeneous thin films could be obtained by spin coating. The dendritic guest-host complexes showed a significantly higher emission upon binding then that of the individual molecules due to the three-dimensional orientation of the OPV guest molecules. In the solid state, this enhancement in luminescence was a factor of 10. The pi-conjugated oligomers are less aggregated in the supramolecular assemblies presumably because of a shielding effect of the bulky adamantyl units present in the hosts. PMID- 14558845 TI - Photohydration and photosolvolysis of biphenyl alkenes and alcohols via biphenyl quinone methide-type intermediates and diarylmethyl carbocations. AB - Evidence is presented for the photochemical generation of novel biphenyl quinone methide (BQM)-type intermediates on photolysis of hydroxybiphenyl alkenes 7 and 8 and hydroxybiphenyl alcohols 9 and 10. Mechanistic investigations utilizing product, fluorescence, and nanosecond laser flash photolysis (LFP) studies indicate two distinct pathways for the formation of these BQMs depending upon the functional groups of the progenitor. Formal excited-state intramolecular proton transfer (ESIPT) between the phenol and the alkene led to BQMs upon irradiation of the hydroxybiphenyl alkenes 7 and 8, while excited-state proton transfer (ESPT) to solvent followed by dehydroxylation was responsible for BQM formation from the hydroxybiphenyl alcohols 9 and 10. Photolysis of 7 and 8 in aqueous CH(3)CN gave photohydration products via attack of water on the respective BQMs, while photolysis of the analogous methyl ethers (of the phenolic moiety) gave only carbocation intermediates. Hydroxybiphenyl alcohols 9 and 10 yielded the corresponding photomethanolysis products in aqueous methanol, through attack of CH(3)OH on the respective BQMs. Although no evidence was found for BQM formation in LFP studies of 8 and 10, due to its suspected short lifetime, the respective diaryl carbocation (lambda(max) 420 nm, tau = 8.5 micros) has been observed upon irradiation of 8 in 2,2,2-trifluoroethanol. A BQM (lambda(max) 580 nm) was observed for 9 but not for 10, the latter having more complex chemistry on laser excitation, resulting in a transient that appears to mask any BQM absorption. Significant quenching of fluorescence from the hydroxybiphenyl alkenes at low water content implies that H(2)O is directly involved in reaction from the singlet excited state. The decrease in fluorescence intensity of 8 was found to depend on [H(2)O](3); however, the distance required for ESIPT in these systems is too large to be bridged by a water trimer. The nonlinear quenching has been attributed to deprotonation of the phenol by two water molecules, with concerted protonation at the alkene by another molecule of water. Fluorescence quenching of the hydroxybiphenyl alcohols required much higher water content, implying a different mechanism of reaction, consistent with the proposal of ESPT (to solvent water) followed by dehydroxylation. PMID- 14558846 TI - Homoleptic cyclometalated iridium complexes with highly efficient red phosphorescence and application to organic light-emitting diode. AB - Phosphorescence studies of a series of facial homoleptic cyclometalated iridium(III) complexes have been carried out. The complexes studied have the general structure Ir(III)(C-N)(3), where (C-N) is a monoanionic cyclometalating ligand: 2-(5-methylthiophen-2-yl)pyridinato, 2-(thiophen-2-yl)-5 trifluoromethylpyridinato, 2,5-di(thiophen-2-yl)pyridinato, 2,5-di(5 methylthiophen-2-yl)pyridinato, 2-(benzo[b]thiophen-2-yl)pyridinato, 2-(9,9 dimethyl-9H-fluoren-2-yl)pyridinato, 1-phenylisoquinolinato, 1-(thiophen-2 yl)isoquinolinato, or 1-(9,9-dimethyl-9H-fluoren-2-yl)isoquinolinato. Luminescence properties of all the complexes at 298 K in toluene are as follows: quantum yields of phosphorescence Phi(p) = 0.08-0.29, emission peaks lambda(max) = 558-652 nm, and emission lifetimes tau = 0.74-4.7 micros. Bathochromic shifts of the Ir(thpy)(3) family [the complexes with 2-(thiophen-2-yl)pyridine derivatives] are observed by introducing appropriate substituents, e.g., methyl, trifluoromethyl, or thiophen-2-yl. However, Phi(p) of the red emissive complexes (lambda(max) > 600 nm) becomes small, caused by a significant decrease of the radiative rate constant, k(r). In contrast, the complexes with the 1 arylisoquinoline ligands are found to have marked red shifts of lambda(max) and very high Phi(p) (0.19-0.26). These complexes are found to possess dominantly (3)MLCT (metal-to-ligand charge transfer) excited states and have k(r) values approximately 1 order of magnitude larger than those of the Ir(thpy)(3) family. An organic light-emitting diode (OLED) device that uses Ir(1 phenylisoquinolinato)(3) as a phosphorescent dopant produces very high efficiency (external quantum efficiency eta(ex) = 10.3% and power efficiency 8.0 lm/W at 100 cd/m(2)) and pure-red emission with 1931 CIE (Commission Internationale de L'Eclairage) chromaticity coordinates (x = 0.68, y = 0.32). PMID- 14558847 TI - Nucleophilic reactivities of carbanions in water: the unique behavior of the malodinitrile anion. AB - The kinetics of the reactions of nine carbanions 1a-i, each stabilized by two acyl, ester, or cyano groups, with benzhydrylium ions in water were investigated photometrically at 20 degrees C. Because the competing reactions of the benzhydrylium ions with water and hydroxide ions are generally slower, the second order rate constants of the reactions of the benzhydrylium ions with the carbanions can be determined with high precision. The rate constants thus obtained can be described by the Ritchie equation, log(k/k(0)) = N(+) (eq 1), which allows us to calculate Ritchie N(+) parameters for a series of stabilized carbanions, for example, malonate, acetoacetate, malodinitrile, etc., and compare them with those of other n-nucleophiles in water (hydroxide, amines, azide, thiolates, etc.). Because the Ritchie relationship (eq 1) is a special case of the more general relationship log k = s(N + E) (eq 4), the reactivity parameters N and s for the carbanions 1a-i can also be calculated and compared with the nucleophilic reactivities of a large variety of n-, pi-, and sigma-nucleophiles, including reactivities of carbanions in dimethyl sulfoxide. While the acyl and ester substituted carbanions are approximately 3 orders of magnitude less reactive in water than in dimethyl sulfoxide, the malodinitrile anion (1i) shows almost the same reactivity in both solvents. Correlations between the nucleophilic reactivities of carbanions with the pK(a) values of the corresponding CH acids reveal that the malodinitrile anion (1i) is considerably more nucleophilic than was expected on the basis of its pK(a) value. This deviation is assigned to the exceptionally low Marcus intrinsic barriers of the reactions of the malodinitrile anion (1i). PMID- 14558848 TI - Why is there an epidemic of myopia? PMID- 14558849 TI - The biological basis of myopic refractive error. AB - Myopia is among the most common refractive errors and is associated with the greatest risk of pathological outcomes. Most animals, including humans, are born with hyperopic errors. During development, axial elongation of the eye occurs and is regulated through a vision-dependent process, known as emmetropisation The extremely rapid changes in the prevalence of myopia and the dependence of myopia on the level of education indicate that there are very strong environmental impacts on the development of myopia. This conflicts with the common occurrence of familial patterns of inheritance of myopia, which suggests a role for genetic determination. There are more than 150 defined genetic syndromes in which familial high myopia is one of the features, including some that are not associated with other syndromes. The evidence for the roles of both nature and nurture in the aetiology of myopia is discussed. This review also examines the experimentally induced refractive errors associated with form-deprivation, recovery from form deprivation and the effects of both negative and positive lenses. In addition, it looks at the local and optical control of eye growth. Finally, the various control pathways for growth are considered. These include dopamine, ZENK-glucagon, retinoic acid and retinoic acid receptors, crystallin, seratonin and melatonin, vasoactive intestinal peptide and enkephalins, nitric oxide and various growth factors. PMID- 14558850 TI - A synopsis of the prevalence rates and environmental risk factors for myopia. AB - The prevalence rates of myopia are higher in urban Asian cities such as Hong Kong and Singapore. One observation over the past few decades is that the prevalence rates of myopia have been rising and there is an epidemic of myopia in Asia. The age-old question of the roles of nature and nurture in this process remains unanswered. The strongest evidence for an environmental link to myopia is near work activity. Childhood exposure to night lighting has also been explored in different studies but the results have been mixed. Twin studies, segregation analysis and association studies have demonstrated that hereditary factors play an important role in myopia development. The exact nature and interplay of genetic and environmental factors is not known and data suggest that environmental factors may interact with genetic factors to increase the risks of developing myopia. Future research is needed to identify specific modifiable lifestyle factors and genetic markers for myopia. This will enable preventive measures such as health education to be instituted. PMID- 14558851 TI - Molecular biology of myopia. AB - Experiments in animal models of myopia have emphasised the importance of visual input in emmetropisation but it is also evident that the development of human myopia is influenced to some degree by genetic factors. Molecular genetic approaches can help to identify both the genes involved in the control of ocular development and the potential targets for pharmacological intervention. This review covers a variety of techniques that are being used to study the molecular biology of myopia. In the first part, we describe techniques used to analyse visually induced changes in gene expression: Northern Blot, polymerase chain reaction (PCR) and real-time PCR to obtain semi-quantitative and quantitative measures of changes in transcription level of a known gene, differential display reverse transcription PCR (DD-RT-PCR) to search for new genes that are controlled by visual input, rapid amplification of 5' cDNA (5'-RACE) to extend the 5' end of sequences that are regulated by visual input, in situ hybridisation to localise the expression of a given gene in a tissue and oligonucleotide microarray assays to simultaneously test visually induced changes in thousands of transcripts in single experiments. In the second part, we describe techniques that are used to localise regions in the genome that contain genes that are involved in the control of eye growth and refractive errors in mice and humans. These include quantitative trait loci (QTL) mapping, exploiting experimental test crosses of mice and transmission disequilibrium tests (TDT) in humans to find chromosomal intervals that harbour genes involved in myopia development. We review several successful applications of this battery of techniques in myopia research. PMID- 14558852 TI - Emmetropisation responses when visual information is presented at only one or two near target planes in chick. AB - PURPOSE: When visual information is confined to one object plane and zero or hyperopic defocus is present, emmetropisation is directed towards this plane, when myopic defocus is present, emmetropisation processes fail. We investigated the effect of introducing information at a second nearer plane on emmetropisation responses under these conditions. METHODS: The visual environment was controlled using lenses (+30 D, +40 D and +50 D) and cones with a Maltese cross target (MX) at one or two distances. Four different target configurations were used: 1. a single target was located at 3.3 cm, 2. a vertical hemi-field target was added at 2.5 cm, 3. a transparent target was added at 2.5 cm or 4. a single target was located at 2.5 cm. An additional cone length of 4.0 cm and nearer target distance of 3.3 cm was used with the +50 D lens. The imaging devices were applied monocularly to eight-day-old chicks and worn for four days. At the end of the treatment, refractive errors and eye growth were measured. Potential regional differences in growth were also assessed. RESULTS: The configuration of the target and the interaction between target configuration and lens power had significant refractive and axial growth effects. With a single target plane, myopic and hyperopic defocus resulted in myopia. When defocus was experienced at two planes, refractive errors shifted towards the plane with the lower defocus and emmetropisation responses, although still not normal, were more consistent. CONCLUSIONS: When visual information is provided at two distances, the target with the lesser incident defocus has the greater influence on the resultant refractive error. Emmetropisation responses are more accurate when information is presented at many distances. PMID- 14558853 TI - Effect of optical defocus on multifocal ERG responses. AB - BACKGROUND: Multifocal electroretinography (mfERG) is a sensitive technique to probe retinal function topographically. Various conditions such as macular degeneration decrease the first-order kernel (K1) response magnitude of mfERG. Previous studies have given inconsistent results on the effect of optical defocus due to poor controls. This study investigated the effect of optical defocus on the K1 response of the central retina using a well-controlled method. METHODS: Twenty subjects were recruited to undergo mfERG measurement using the VERIS 4.0 system. A four millimetre artificial pupil was fitted before each fully-dilated right eye, optically corrected for the viewing distance. The implicit times and response amplitudes of n1 (first negative trough) and p1 (first positive peak) under three different optical defocus conditions (zero dioptres, +1.00 D and +3.00 D) were measured. RESULTS: The implicit times of n1 and p1 did not demonstrate any significant variation from the central macula to para-macula under different optical defocus conditions. The response amplitude of n1 did not show any changes from the central macula to para-macula but the response amplitude of the central macular p1 showed a significant reduction by 12 per cent under +1.00 D defocus and +3.00 D defocus. CONCLUSIONS: Optical defocus causes mild reduction in mfERG at the central macula but there are no significant changes in the periphery. A full optical correction is recommended for mfERG measurement to minimise the reduction of the macular response due to optical defocus. PMID- 14558854 TI - AC/A ratios in myopic and emmetropic Hong Kong children and the effect of timolol. AB - PURPOSE: Caucasian children with myopia have elevated response accommodative vergence to accommodation (AC/A) ratios. The purpose of this study was twofold: to determine if response AC/A ratios vary with refractive error and with myopic progression rate in Hong Kong Chinese children, and to determine the effect of beta-adrenergic antagonism with topical timolol application on AC/A ratios. METHODS: Thirty children aged eight to 12 years participated in the study. All refractive errors were corrected with spectacle lenses. Accommodative responses were measured using a Shin-Nippon autorefractor and concurrent changes in vergence were assessed using a vertical prism and a Howell-Dwyer card at three metres and 0.33 metre. Accommodative demand was altered using plus or minus two dioptre lenses and lens- and distance-induced response AC/A ratios were calculated. Measurements were repeated 30 minutes after the instillation of topical timolol maleate (0.5 per cent). RESULTS: AC/A ratios appeared higher in progressing myopic children but the difference was not statistically significant. Timolol application reduced accommodative convergence (AC) in the stable myopes (reduction = -3 +/- 1.14 prism dioptres) but not in the emmetropes (0.69 +/- 0.96 prism dioptres) or progressing myopes (0.16 +/- 0.43 prism dioptres) and this difference between refractive groups was statistically significant (F(2, 27) = 3.766; P = 0.036). However, timolol did not produce a significant change in the accommodative response to positive or negative lenses or response AC/A ratios. CONCLUSIONS: We did not find that AC/A ratios in myopic Chinese children were elevated and therefore, it is unlikely that elevated AC/A ratios are responsible for the high levels of myopia that occur in Hong Kong. The finding that timolol reduced AC in the stable myopes suggests that the autonomic control of accommodative convergence in these children may be different from that in emmetropic children and those with progressing myopia. PMID- 14558855 TI - An assessment of consecutively presenting orthokeratology patients in a Hong Kong based private practice. AB - PURPOSE: The aim of this study was to collect objective, subjective and demographic data on consecutively presenting orthokeratology (ortho-k) patients who attended for routine follow-up examination in a Hong Kong based private practice in May 2001. METHOD: Sixty-nine patients who returned to the surveyed practice for follow-up visits during the study period (May 2001) were interviewed and relevant data extracted from their files. Data collected included identification and estimation of the extent of complications encountered by ortho k patients and their satisfaction with the treatment. RESULTS: Among the 61 patients who had been wearing ortho-k lenses for at least one month, 50 patients were younger than 16 years old. Twelve children (24 per cent) had been reluctant to wear ortho-k lenses before undergoing the treatment but, after commencement of lens wear, only one child was not very willing to wear the ortho-k lenses. The mean pre-ortho-k spherical refractive error of these patients was -3.93 +/- 2.30 D (OS only). Of the 59 patients who wore ortho-k lenses for at least one month and who were on night therapy, 10 patients had to wear spectacles or contact lenses in the daytime due to significant residual myopia. There was no statistically significant correlation between post-ortho-k unaided visual acuity and pre-ortho-k refractive error (spherical, cylindrical or the equivalent sphere) in the 49 patients who did not need to wear any vision correction in the daytime. Of the 61 patients, four reported eye inflammation/infection during the treatment. All recovered their ocular health without any effect on their vision or corneal health. The incidence of corneal staining that required lens wear to be stopped appeared to increase with the duration of ortho-k lens treatment. The incidence of staining was not related to refractive error, unaided visual acuity or the age of the subjects. The most common problem reported by the patients was lens binding and there were also reports of increased redness, itching, light sensitivity and secretion of mucus in the morning after opening their eyes. More than 50 per cent of the patients experienced some distance vision blur, which was worse towards the end of the day. For most patients, these problems occurred only occasionally. Higher pre-ortho-k spherical refractive error was related to poor near and distance vision and worse distance vision towards the end of the day. CONCLUSION: The majority of the patients interviewed were children who reported being 'happy with the results of the treatment'. Night wear is the main wearing modality and in view of the increased risk of complications in overnight wear and the fact that a large number of the patients are children, the need for strict compliance with the practitioner's instructions for lens use and care cannot be overemphasised. With careful monitoring and good compliance, complications with overnight ortho-k wear can be minimised. In view of the high incidence of lens binding, it is essential that patients and parents of young patients know the correct method to free a bound lens. PMID- 14558856 TI - Prevalence of myopia among primary school children in eastern Sydney. AB - BACKGROUND: Worldwide there is concern that the prevalence of myopia is increasing but the prevalence of myopia in Australian school children has not been analysed in detail. This study examines the prevalence of refractive errors in a large unselected population of primary school children in the eastern suburbs of Sydney. METHODS: Children visiting the Vision Education Centre at the School of Optometry and Vision Science, University of New South Wales, on a school excursion during the first half of the 1990s were refracted by non cycloplegic retinoscopy. Spherical equivalents were computed and analysis of variance carried out. RESULTS: There were 1,459 boys and 1,076 girls in the study, a total of 2,535 children of whom approximately 40 per cent were born overseas or were first generation Australians. Overall, there was no significant difference in refractive error between girls and boys, although there were age/gender interactions with older girls exhibiting more hyperopia than boys. The refractive error ranged from -0.25 to +1.25 D spherical equivalent in 87.3 per cent of children. Mean spherical equivalent for the group was +0.50 +/- 0.82 D. There was a significant shift (p < 0.001) towards increasing myopia with age. The prevalence of myopia greater than -0.50 D rose from 1.0 per cent of four-year olds to 8.3 per cent of 12-year-olds. CONCLUSIONS: The prevalence of myopia found in a large multi-ethnic group of non-clinical primary school children in eastern Sydney is lower than expected from other studies in the USA and Asia. Compared with Australian data from the 1970s and 1980s, only a weak increase in the prevalence of myopia is revealed. PMID- 14558857 TI - Theodore P Grosvenor. Foundation head of the optometry course at the University of Auckland. PMID- 14558858 TI - Evidence and information for health policy: a decade of change. PMID- 14558859 TI - Coax, COX and cola. PMID- 14558860 TI - Substance use, psychological distress and crime. PMID- 14558861 TI - Does every baby get a newborn screening test? PMID- 14558862 TI - Lessons from early large-scale adoption of celecoxib and rofecoxib by Australian general practitioners. AB - OBJECTIVE: To assess trends in the first two years of prescribing of COX-2 selective non-steroidal anti-inflammatory drugs (C2SNs) by Australian general practitioners. DESIGN: Retrospective analysis of deidentified electronic patient records from GPs enrolled in the General Practice Research Network (GPRN). SETTING AND PARTICIPANTS: Overall prescription rates for C2SNs and NSAIDs were assessed for all GPRN participants (437 GPs) between 1 September 1999 and 30 September 2002. Also, three cohorts of patients, with at least 12 months of prescription data, who received their first prescription for celecoxib between August and October 2000 (Cohort 1, 2366 patients), celecoxib between February and April 2001 (Cohort 2, 640 patients), and rofecoxib between February and April 2001 (Cohort 3, 608 patients) were selected for further analysis. MAIN OUTCOME MEASURES: Age and sex of patients; reason for prescription; previously prescribed pain medications and concomitant use of medications that could predispose to an adverse renal or bleeding event. RESULTS: Prescriptions for C2SNs increased dramatically after they were listed on the Pharmaceutical Benefits Scheme (PBS). C2SN prescriptions for patients aged less than 65 years accounted for 52.6%, 59.5% and 50.7% of those in Cohorts 1, 2 and 3, respectively; large numbers of patients in the study cohort had reasons recorded for prescription that did not comply with PBS restrictions, and between 36.7% and 61.3% of patients in the three cohorts had not received a prescription for any pain medication in the year before being prescribed a C2SN. Between 4.7% and 7.9% were coprescribed drugs that could cause renal complications. CONCLUSIONS: Rapid, early adoption of C2SNs by Australian GPs has resulted in prescribing and drug use patterns that were not in accord with quality use of medicine (QUM) principles. PMID- 14558863 TI - Substance-use disorders and psychological distress among police arrestees. AB - OBJECTIVES: To determine the 12-month prevalence of substance-use disorders and psychological morbidity in an Australian arrestee population. DESIGN: Cross sectional descriptive study. PARTICIPANTS AND SETTING: 288 police arrestees at the Brisbane City Police Watch House in February and March 2001. OUTCOME MEASURES: Prevalence of drug and alcohol disorders; psychological "caseness" according to the 28-item General Health Questionnaire; demographics and index offences. RESULTS: 86% of the arrestees had at least one substance-use disorder; most had multiple disorders. More than 80% were substance dependent. The predominant substances used were amphetamines, marijuana, opioids and alcohol. 82% of the men and 94% of the women were suffering significant psychological distress. CONCLUSIONS: Development of services for detoxification and treatment of this population is a pressing need. The findings provide crucial information for the planning and implementation of drug courts and court diversion systems. PMID- 14558864 TI - Newborn screening in South Australia: is it universal? AB - OBJECTIVE: To determine the biochemical screening rate of newborns in South Australia and the factors associated with babies not being screened. DESIGN: Matching of data in the SA Newborn Screening Centre database (acquired from Guthrie cards) with the SA perinatal data collection (compiled from supplementary birth records) to determine how many newborns missed screening. Risk factors for missed screening were identified from sociodemographic and clinical variables recorded in the perinatal data collection and analysed by multivariable unconditional logistic regression analysis. PATIENTS AND SETTING: All live births (n = 18,426) in South Australia in 1999, in the 63 hospitals assisting deliveries or in the home. MAIN OUTCOME MEASURES: Rates of biochemical screening and missed screening in all newborns and among various subgroups; adjusted odds ratios (after multivariable logistic regression analysis) for risk factors for missed screening. RESULTS: The newborn screening rate in South Australia in 1999 was 97.8%. Babies born at home, born to an Aboriginal mother, or born to a mother who normally resided in another state were at higher risk of missed screening. Other factors associated with missed screening were having fewer than seven antenatal visits, prematurity (gestational age at birth < 32 weeks), congenital abnormality in the baby, use of paediatric intensive care, early discharge from hospital before 3 days (but especially after less than 1 day), and death of the baby during the neonatal period. CONCLUSION: In South Australia, while 2.2% of all newborns missed screening in 1999, in certain high-risk groups the proportions of unscreened babies were significantly higher. With a 2% missed screening rate, one might expect one newborn with a screening-detectable disorder to elude detection every other year in South Australia. PMID- 14558865 TI - The influence of geographical location on the complexity of rural general practice activities. AB - OBJECTIVES: To examine the complexity of activities undertaken in general practice in relation to degree of rurality of the practice. DESIGN AND SETTING: National mail questionnaire survey across non-metropolitan Australia in July 2002. PARTICIPANTS: 1498 respondents out of 4406 GPs providing at least 375 Medicare-rebatable consultations in rural and remote locations during January March 2002 (response rate, 35%). MAIN OUTCOME MEASURES: Responses to five sentinel measures of practice complexity. RESULTS: In general, the proportion of GPs providing complex services increases with increasing rurality or remoteness. Isolated rural and remote GPs manage myocardial infarctions to a higher level than GPs in larger rural and regional centres, are more likely to administer cytotoxic drugs, perform forensic examinations, stabilise injured patients pending retrieval, and coordinate discharge planning more often. CONCLUSIONS: The more rural or remote the area, the more likely a GP is to be regularly engaged in complex care. These findings have implications for the workload, responsibility, vocational satisfaction, need for professional education and support, and costs and remuneration of practice. PMID- 14558866 TI - Heart failure: how can we prevent the epidemic? AB - Heart failure prevalence is increasing because of the ageing of the population and the longer survival of people experiencing myocardial infarction and heart failure. The lifetime risk of developing heart failure in Western countries is about 20%. The increasing prevalence of overweight, obesity and diabetes is likely to accelerate heart failure incidence. While there have been major advances in treating heart failure, a preventive approach promises greater benefit to a larger proportion of the community. The medical strategy for heart failure prevention, based on calculation of individual risk, is focused on the minority of individuals who exceed an arbitrary risk threshold. A public health strategy targeting the whole population offers a greater prospect of reducing the incidence of heart failure and other cardiovascular disease. A multitiered approach, encompassing environmental determinants of lifestyle, legislation, and education about healthy lifestyles throughout life, in addition to aggressive control of risk factors in high-risk individuals, is likely to have the greatest impact. PMID- 14558867 TI - Adverse event reporting in clinical trials: room for improvement. AB - Regulatory and ethical guidelines require clinical trial sponsors to disseminate clinical trial adverse event reports to involved investigators and human research ethics committees. Compliance with these guidelines has resulted in a major administrative burden for ethics committees. This burden does not necessarily contribute to the protection of clinical trial participants. Rationalisation of the adverse event reporting might allow better use of the data and might benefit human research ethics committees. PMID- 14558868 TI - First report of human angiostrongyliasis acquired in Sydney. PMID- 14558869 TI - Money, morals and the conquest of mortality. PMID- 14558870 TI - Is early surgical referral for children with persistent otitis media with effusion (OME) appropriate? PMID- 14558871 TI - Inclusion of patients in clinical trial analysis: the intention-to-treat principle. PMID- 14558872 TI - 2: Recent advances in therapy of diabetes. AB - As suboptimal blood glucose control has a lasting harmful effect even if control improves later, intensive insulin therapy to minimise hyperglycaemia is now recommended for all patients with type 1 diabetes. The new rapid- and long-acting insulin analogues offer more physiological insulin profiles than traditional insulin preparations. Continuous insulin infusion ("pump therapy") may provide a solution for some patients with frequent hypoglycaemia or hypoglycaemic unawareness. Continuous blood glucose monitoring reveals postprandial hyperglycaemia and asymptomatic nocturnal hypoglycaemia and may be especially useful for programming overnight basal insulin rates for pump therapy. In type 2 diabetes, management should change with disease progression; introduction of insulin should not be delayed if metabolic control becomes suboptimal. PMID- 14558874 TI - Energy levels for biphasic defibrillation. PMID- 14558875 TI - Tasmania: doing its wee bit for iodine nutrition. PMID- 14558876 TI - Leprosy transmission in the Kimberley, Western Australia: still a reality in 21st century Australia. PMID- 14558877 TI - Computerised asthma action plans. PMID- 14558878 TI - Mirtazapine-induced hyponatraemia. PMID- 14558879 TI - New contraceptive choices across reproductive life. PMID- 14558880 TI - Update on treatment of menstrual disorders. PMID- 14558881 TI - Latrodectism: a prospective cohort study of bites by formally identified redback spiders. PMID- 14558882 TI - Breast self-examination: be alert but not alarmed? PMID- 14558883 TI - Interaction of a farnesylated protein with renal type IIa Na/Pi co-transporter in response to parathyroid hormone and dietary phosphate. AB - Treatment with PTH (parathyroid hormone) or a high-P(i) diet causes internalization of the type IIa sodium-dependent phosphate (Na/P(i) IIa) co transporter from the apical membrane and its degradation in the lysosome. A dibasic amino acid motif (KR) in the third intracellular loop of the co transporter is essential for protein's PTH-induced retrieval. To elucidate the mechanism of internalization of Na/P(i) IIa, we identified the interacting protein for the endocytic motif by yeast two-hybrid screening. We found a strong interaction of the Na/P(i) IIa co-transporter with a small protein known as the PEX19 (human peroxisomal farnesylated protein; PxF, Pex19p). PEX19 can bind to the KR motif, but not to a mutant with this motif replaced with NI residues. PEX19 is highly expressed in mouse and rat kidney. Western blot analysis indicates that PEX19 is located in the cytosolic and brush-border membrane fractions (microvilli and the subapical component). Overexpression of PEX19 stimulated the endocytosis of the Na/P(i) IIa co-transporter in opossum kidney cells in the absence of PTH. In conclusion, the present study indicates that PEX19 may be actively involved in controlling the internalization and trafficking of the Na/P(i) IIa co-transporter. PMID- 14558884 TI - Influence of a mutation in the ATP-binding region of Ca2+/calmodulin-dependent protein kinase II on its interaction with peptide substrates. AB - CaMKII (Ca2+/calmodulin-dependent protein kinase II) is expressed in high concentrations in the brain and is found enriched in the postsynaptic densities. The enzyme is activated by the binding of calmodulin to the autoregulatory domain in the presence of high levels of intracellular Ca2+, which causes removal of auto-inhibition from the N-terminal catalytic domain. Knowledge of the 3D (three dimensional) structure of this enzyme at atomic resolution is restricted to the association domain, a region at the extreme C-terminus. The catalytic domain of CaMKII shares high sequence similarity with CaMKI. The 3D structure of the catalytic core of CaMKI comprises ATP- and substrate-binding regions in a cleft between two distinct lobes, similar to the structures of all protein kinases solved to date. Mutation of Glu-60, a residue in the ATP-binding region of CaMKII, to glycine exerts different effects on phosphorylation of two peptide substrates, syntide and NR2B ( N -methyl-D-aspartate receptor subunit 2B) 17-mer. Although the mutation caused increases in the Km values for phosphorylation for both the peptide substrates, the effect on the kcat values for each was different. The kcat value decreased in the case of syntide, whereas it increased in the case of the NR2B peptide as a result of the mutation. This resulted in a significant decrease in the apparent kcat/Km value for syntide, but the change was minimal for the NR2B peptide. These results indicate that different catalytic mechanisms are employed by the kinase for the two peptides. Molecular modelling suggests structural changes are likely to occur at the peptide-binding pocket in the active state of the enzyme as a consequence of the Glu-60-->Gly mutation. PMID- 14558885 TI - Benefits of early enteral nutrition with glutamine and probiotics in brain injury patients. AB - Brain injury patients have higher energy and protein expenditures and are prone to infections. The aim of the present study was to evaluate the results of early enteral feeding with glutamine and probiotics in brain injury patients. Twenty three brain injury patients (Glasgow score between 5-12 and therapeutic intervention scoring system>20) were studied. Three patients were excluded to leave 20 remaining patients. Patients were randomized to receive either an early enteral diet (control group, n=10) or the same formula with glutamine and probiotics added (study group, n=10) for a minimum of 5 days (range, 5-14 days). The diets were isocaloric and isonitrogenous [35 kcal.kg(-1).day(-1) (where 1 kcal approximately 4.184 kJ) and 1.5 g of protein.kg(-1).day(-1)]. Main outcome measures were the incidence of infection, the length of stay in the intensive care unit and the number of days requiring mechanical ventilation. The two groups were homogeneous in gender, age, nutritional status and severity of trauma. There was no mortality during the study period. The infection rate was higher in controls (100%) when compared with the study group (50%; P=0.03) and the median (range) number of infections per patient was significantly greater (P<0.01) in the control group [3 (1-5)] compared with the study group [1 (0-3)]. Both the critical care unit stay [22 (7-57) compared with 10 (5-20) days; P<0.01; median (range)] and days of mechanical ventilation [14 (3-53) compared with 7 (1-15) days; P=0.04; median (range)] were higher in the patients in the control group than in the study group. We conclude that the enteral formula containing glutamine and probiotics decreased the infection rate and shortened the stay in the intensive care unit of brain injury patients. PMID- 14558888 TI - Pathways for protection from noise induced hearing loss. AB - There is increasing evidence that at least one function of both the medial and the lateral olivocochlear efferent systems is to provide adjustment of the set point of activity in their postsynaptic target, the outer hair cells and afferent processes, respectively. New results, summarized in this review, suggest that both efferent systems can provide protection from noise through this mechanism. There are also intracellular pathways that can provide protection from noise induced cellular damage in the cochlea. This review also summarizes new results on the pathways that regulate and react to levels of reactive oxygen species in the cochlea as well as the role of stress pathways for the heat shock proteins and for neurotrophic factors in protection, recovery and repair. PMID- 14558887 TI - The heptapeptide LSARLAF mediates platelet activation through phospholipase Cgamma2 independently of glycoprotein IIb-IIIa. AB - The seven-amino-acid peptide LSARLAF has been reported to activate platelets via the integrin GPIIb-IIIa (glycoprotein IIb-IIIa). Activation by LSARLAF is reinforced by release of ADP and thromboxanes, but the initiating event in the signalling cascade is not known. In the present study, we demonstrate that LSARLAF stimulates Src kinase-dependent tyrosine phosphorylation of many of the proteins in the GPIIb-IIIa cascade, including the tyrosine kinase Syk, the adapter SLP-76 (SH2-containing leucocyte phosphoprotein of 76 kDa) and PLCgamma2 (phospholipase Cgamma2). A critical role for PLCgamma2 in signalling by LSARLAF was demonstrated by abolition of aggregation in PLCgamma2-/- murine platelets to low concentrations of the peptide, although a partial recovery was seen with higher concentrations. In sharp contrast with the GPIIb-IIIa-regulated signalling cascade, aggregation was inhibited in murine platelets deficient in the adapter LAT (linker for activation of T-cells) and the Fc receptor gamma-chain. Aggregation was also partially inhibited by the cholesterol-lowering reagent, beta-methyl-cyclodextrin, at concentrations that disrupt membrane rafts, but do not interfere with signalling by GPIIb-IIIa. Furthermore, LSARLAF also stimulated tyrosine phosphorylation in GPIIb-deficient murine platelets, confirming that the integrin is not critical for activation of intracellular signalling pathways. LSARLAF also stimulated Ca2+ elevation in RBL-2H3 cells, which lack the platelet glycoproteins GPIIb, GPVI and GPIb. These results demonstrate that LSARLAF activates platelets through a PLCgamma2-dependent pathway that lies downstream of Src kinases and which is partially dependent on the Fc receptor gamma-chain, LAT and lipid rafts. The mechanism of cell activation by LSARLAF remains to be established, although the present results indicate that more than one surface glycoprotein may mediate this response. PMID- 14558889 TI - Genetic influences in individual susceptibility to noise: a review. AB - Individual animals and humans show differing susceptibility to noise damage even under very carefully controlled exposure conditions. This difference in susceptibility may be related to unknown genetic components. Common experimental animals (rats, guinea pigs, chinchillas, cats) are outbred-their genomes contain an admixture of many genes. Many mouse strains have been inbred over many generations reducing individual variability, making them ideal candidates for studying the genetic modulation of individual susceptibility. Erway et al. (1993) demonstrated a recessive gene associated with early presbycusis in the C57BL/6J inbred mouse. A series of studies have shown that mice homozygous for Ahl allele are more sensitive to the damaging effects of noise. Recent work has shown that mice homozygous for Ahl are not only more sensitive to noise, but also are probably damaged in a different manner by noise than mice containing the wild type gene (Davis et al., 2001). Recent work in Noben-Trauth's lab (Di Palma et al., 2001) has shown that the wild-type Ahl gene codes for a hair cell specific cadherin. Cadherins are calcium dependent proteins that hold cells together at adherins junctions to form tissues and organs. The cadherin of interest named otocadherin or CDH23, is localized to the stereocillia of the outer hair cells. Our working hypothesis, suggests that otocadherin may form the lateral links between stereocilia described by Pickles et al (1989). Reduction of, or missing otocadherin weakens the cell and may allow stereocilia to be more easily physically damaged by loud sounds and by aging. PMID- 14558886 TI - Modification of phospholipase C-gamma-induced Ca2+ signal generation by 2 aminoethoxydiphenyl borate. AB - The mechanisms by which Ca(2+)-store-release channels and Ca(2+)-entry channels are coupled to receptor activation are poorly understood. Modification of Ca(2+) signals by 2-aminoethoxydiphenyl borate (2-APB), suggests the agent may target entry channels or the machinery controlling their activation. In DT40 B-cells and Jurkat T-cells, complete Ca(2+) store release was induced by 2-APB (EC(50) 10-20 microM). At 75 microM, 2-APB emptied stores completely in both lymphocyte lines, but had no such effect on other cells. In DT40 cells, 2-APB mimicked B-cell receptor (BCR) cross-linking, but no effect was observed in mutant DT40 lines devoid of inositol 1,4,5-trisphosphate (InsP(3)) receptors (InsP(3)Rs) or phospholipase C-gamma2 (PLC-gamma2). Like the BCR, 2-APB activated transfected TRPC3 (canonical transient receptor potential) channels, which acted as sensors for PLC-gamma2-generated diacylglycerol in DT40 cells. The action of 2-APB on InsP(3)Rs and TRPC3 channels was prevented by PLC-inhibition, and required PLC gamma2 catalytic activity. However, unlike BCR activation, no increased InsP(3) level could be measured in response to 2-APB. Also, calyculin A-induced cytoskeletal reorganization prevented 2-APB-induced InsP(3)R and TRPC3-channel activation, but not that induced by the BCR. 2-APB still activated TRPC3 channels in DT40 cells with fully depleted Ca(2+) stores, indicating its action was not via Ca(2+) release. Significantly, 2-APB-induced InsP(3)R and TRPC3 activation was prevented in DT40 knockout cells devoid of the BCR- and PLC-gamma2-coupled adaptor/kinases, Syk, Lyn, Btk or BLNK. The results suggest that 2-APB activates Ca(2+) signals in lymphocytes by initiating and enhancing coupling between components of the BCR-PLC-gamma2 complex and both Ca(2+)-entry and Ca(2+)-release channels. PMID- 14558890 TI - Stress in hearing and balance in Meniere's disease. AB - Stress is an unavoidable every-day phenomenon. Physiological coping with stress depends on the appropriate release of stress hormones as well as their alleviation at the termination of the stress. Despite quite a body of research indicating that stress affects inner ear function, this concept has found little application in otolaryngology. Today's evidence clearly indicates that the inner ear is equipped to detect stress hormones and some of these hormones have been shown to affect the inner ear function. Major stress control pathways shown to affect the inner ear include several third order axes, the hypothalamus-pituitary adrenal axis, the hypothalamus-pituitary-thyroid axis and the hypothalamus pituitary-gonadal axis whose functioning are interactive and inter-dependent. Less well-studied are the second order hypothalamus-pituitary control axis and its interaction with other hormones. To explore these we carried out a retrospective study on a series of Meniere's patients who had undergone a neurotomy of the vestibular nerve in the dept of ORL at the Hopital Nord, Marseille. Meniere's patients were particularly appropriate for this study since stress has long been recognised as a factor associated with the triggering of the symptoms of this pathology. Patients with acoustic neuroma and facial spasm were taken as a control population. We investigated the level of a battery of stress hormones including prolactin beta-endorphin and growth hormone. The blood sample was taken on the morning before surgery. The most striking observation was the presence of hyperprolactinemia in 30% of the Meniere patients (more than 20 microg/l) with confirmation of prolactinoma in 6 patients. The level of beta endorphin could also be elevated. Horner, K.C., Guieu, R., Magnan, J., Chays, A. and Cazal, Y. Neuropysychopharmacology, (2001) 26:135-138. These observations suggest that neuroendocrinological feedback pathways controlling stress can be disturbed in Meniere's patients and depression of hypothalamic dopaminergic inhibition of prolactin secretion might be implicated. A further study on non operated Meniere's patients presenting hyperprolactinemia and on dopamine agonist treatment, is needed in order to assess the role of stress in Meniere's patients. Progress in this domain could open the door towards integration of the stress concept into clinical management of various inner ear disorders. PMID- 14558891 TI - A descriptive cross-sectional study of annoyance from low frequency noise installations in an urban environment. AB - In order to improve the living conditions for respondents highly exposed to traffic noise, it has been recommended that one side of the building should face a "quiet side". Quiet may, however, be spoilt by noise from installations such as ventilation and air-conditioning systems. The noises generated by installations of this kind often have a dominant portion of low frequencies (20-200 Hz) and may be a source of great annoyance and sleep disturbance. This paper describes the cross-sectional part of an intended intervention study among residents exposed to traffic noise on one side of the building and to low frequency noise from installations on the other side of the building. A questionnaire masked as a general living environment study was delivered to a randomly selected person in each household. In total 41 respondents answered the questionnaire (71% response rate). Noise from installations was measured indoors in a bedroom facing the courtyard in a selection of apartments and outdoors in the yard. 24h traffic noise outdoor and indoor levels were calculated. The noise levels from installations were slightly above or at the Swedish recommendations for low frequency noise indoors with the window closed and exceeded the recommendations by about 10 dB SPL when the window was slightly opened. The proportion of persons who reported that they were very or extremely annoyed indoors from noise from installations was more than twice as high as for traffic noise. Installation noise also affected respondents' willingness to have their windows open and to sleep with an open window. The high disturbance of installation noises found in this study indicates the importance of also regulating the noise exposure on the "quiet side" of buildings. Further studies will give a better base for the extent of annoyance and acceptable levels of installation noises. PMID- 14558892 TI - The influence of ageing on noise-induced hearing loss. AB - A lifetime of exposure to noise is likely to have negative effects on the hearing, but the interaction between noise-induced hearing loss (NIHL) and age related hearing loss is difficult to determine. The most commonly accepted assumption is a simple accumulating effects of noise and ageing on the hearing. However, both a less than additive effect as well as a supraadditive effect has been proposed. Recently an interesting interaction between NIHL and age-related hearing loss has been reported (Gates et al., 2000). NIHL before old age reduces the effects of ageing at noise-associated frequencies, but accelerates the deterioration of hearing in adjacent frequencies. Findings from the longitudinal and cross-sectional gerontological and geriatric population study of 70-year-olds in Gothenburg, Sweden supports these observations. The incidence of tinnitus increases in old age, but not at the same high rate as presbyacusis. According to the gerontological and geriatric population study in Gothenburg tinnitus in old age is related more to hearing loss than to ageing. There are no simple correlations between exposure to noise during the active years and tinnitus in old age. PMID- 14558893 TI - Is there an association between noise exposure and King Kopetzky Syndrome? AB - The condition in which individuals with normal pure tone audiograms complain of hearing difficulties, especially in the presence of background noise, (normal pure tone audiograms), has had a number of different names. The present term King Kopetzky Syndrome was coined by Hinchcliffe in 1992. This is a common condition reported in 5 - 10% of those attending clinics complaining of hearing problems. A dominant genetic aetiology has been found in a proportion of cases. It may be associated with minor peripheral or central auditory dysfunction, and frequently the individuals exhibit anxious or depressive personalities. We found no relationship with noise exposure in a series of patients compared with matched controls. Here we review the evidence for and against such an influence and present fresh data in an attempt to define the role of noise, if any, in the causation of this condition. Our final conclusion is that there is no clear association between KKS and noise exposure PMID- 14558894 TI - Impulse noise and risk criteria. AB - Impulse noise causes evidently more severe hearing loss than steady state noise. The additional effect of occupational impulse noise on hearing has been shown to be from 5 to 12 dB at 4 kHz audiometric frequency. Reported cases for compensated for hearing loss are prevalent in occupations where noise is impulsive. For impulse noise two measurement methods have been proposed: the peak level method and energy evaluation method. The applicability of the peak level method is difficult as even the recurrent impulses have different time and frequency characteristics. Various national risk criteria differ from international risk criteria. In France the maximum A-weighted peak level is 135 dB, and in the United Kingdom the C-weighted peak sound pressure is limited to 200 Pa (140 dB). This criterion of unweighted 200 Pa (140 dB) is used in European Union (EU) directive 86/188 and ISO 1999-1990 regardless of the number of impulses. The American Conference of Governmental Industrial Hygienists (ACGIH) has recommended that no exposure in excess of a C-weighted peak sound pressure level of 140 dB should be permitted. At work places these norms do not cause any practical consequences since the impulses seldom exceed 140 dB peak level. In several occupations the impulses are so rapid that they contribute only a minimal amount to the energy content of noise. These impulses can damage the inner ear even though they cause reduced awareness of the hazard of noise. Based to the present knowledge it is evident that there is the inadequacy of the equal energy principle in modelling the risk for hearing loss. The hearing protectors attenuate industrial impulse noise effectively due to the high frequency contents of impulses. Directive regarding the exposure of workers to the risks arising from noise requires that in risk assessment attention should be paid also to impulsive noise. So far there is no valid method to combine steady state and impulse noise. A statistical method for the measurements of industrial impulse noise is needed to get a preferably single number for risk assessment. There is an urgent task to develop risk assessment method and risk criteria for impulsive noise to meet the requirements of the upcoming European Union noise directive. PMID- 14558895 TI - Temporary hearing threshold shifts and restitution after energy-equivalent exposures to industrial noise and classical music. AB - In order to investigate whether the energy-equivalence principle is at least acceptable for exposures with a duration in the range of hours and in order to disclose the actual physiological responses to exposures which varied with respect to the time structure and the semantic quality of sounds, a series of tests was carried out where physiological costs associated with varying exposures were measured audiometrically. In a cross-over test design, 10 Subjects (Ss) participated in test series with 3 energetically equal sound exposures on different days. The exposures corresponded with a tolerable rating level of 85 dB / 8 h. In a first test series (TS I), the Ss were exposed to a prototype of industrial noise with a sound pressure level of 94 dB(A) / 1 h. In a second test series (TS II), the same type of noise was applied, but the exposure time of a reduced level of 91 dB(A) was increased to 2 hours. In a third test series (TS III), classical music was provided also for 2 h at a mean level of 91 dB(A). The physiological responses to the 3 exposures were recorded audiometrically via the temporary threshold shift TTS2, the restitution time t(0 dB), and the IRTTS value. IRTTS is the integrated restitution temporary threshold shift which is calculated by the sum of all threshold shifts. It represents the total physiological costs the hearing must "pay" for the sound exposure. Physiological responses of the hearing to the industrial noise exposures in TS I and TS II, all in all, were identical in the 3 parameters. Maximum threshold shifts of approximately 25 dB occurred which did not dissipate completely until 2.5 h after the end of the exposure and IRTTS-values of about 800 dBmin were calculated. Therefore, at least for exposure times in the range of hours, the equilibration of intensity and duration of sound exposures according to the energy-equivalence principle seems to have no influence on the hearing. Classical music was associated with the least severe TTS of less than 10 dB which disappeared much more quickly. IRTTS added up to just about 100 dBmin and, in comparison with 800 dBmin as specific responses to industrial noise, amounted to only about 12%. The substantially lower physiological costs of classical music apparently indicate a decisive influence of the type of sound exposures. Making inferences from the results of the study, the conventional approach of rating sound exposures exclusively by the principle of energy equivalence can lead to gravely misleading assessments of their actual physiological costs. PMID- 14558897 TI - Palaeoclimatic insights into future climate challenges. AB - Palaeoclimatic data document a sensitive climate system subject to large and perhaps difficult-to-predict abrupt changes. These data suggest that neither the sensitivity nor the variability of the climate are fully captured in some climate change projections, such as the Intergovernmental Panel on Climate Change (IPCC) Summary for Policymakers. Because larger, faster and less-expected climate changes can cause more problems for economies and ecosystems, the palaeoclimatic data suggest the hypothesis that the future may be more challenging than anticipated in ongoing policy making. Large changes have occurred repeatedly with little net forcing. Increasing carbon dioxide concentration appears to have globalized deglacial warming, with climate sensitivity near the upper end of values from general circulation models (GCMs) used to project human-enhanced greenhouse warming; data from the warm Cretaceous period suggest a similarly high climate sensitivity to CO(2). Abrupt climate changes of the most recent glacial interglacial cycle occurred during warm as well as cold times, linked especially to changing North Atlantic freshwater fluxes. GCMs typically project greenhouse gas-induced North Atlantic freshening and circulation changes with notable but not extreme consequences; however, such models often underestimate the magnitude, speed or extent of past changes. Targeted research to assess model uncertainties would help to test these hypotheses. PMID- 14558898 TI - Evidence for abrupt climate changes in annually laminated marine sediments. AB - Annually laminated sediments from marine or lacustrine settings represent valuable high-resolution archives of climate change that record variation due to changing precipitation and run-off from land or variation in biological productivity and flux in the water column. Because of their annual resolution such sediments may capture abrupt changes of interannual to decadal scales rivaling corals and ice cores in resolution. Laminated sediments often occur intermittently in the sediment column, and the onset and cessation of laminae commonly record the abrupt crossing of thresholds related to climate change, for example, in the degree of oxygenation of bottom waters. Such records from marginal basins and continental margins have been pivotal in demonstrating that abrupt changes hitherto documented only in high-latitude ice cores are synchronous with climatic change at low latitudes. These insights into global teleconnections have improved our understanding of the mechanisms of rapid climate change. In deep-sea settings, the discovery of the episodic occurrence of laminated diatom-rich sediments in the Equatorial Pacific and Southern Ocean provides evidence for massive climate-related biogeochemical excursions tied to abrupt changes in the input, distribution and availability of nutrients in the oceans. PMID- 14558899 TI - Glacial integrative modelling. AB - Understanding the mechanisms of past climate changes requires modelling of the complex interaction between all major components of the Earth system: atmosphere, ocean, cryosphere, lithosphere and biosphere. This paper reviews attempts at such an integrative approach to modelling climate changes during the glacial age. In particular, the roles of different factors in shaping glacial climate are compared based on the results of simulations with an Earth-system model of intermediate complexity, CLIMBER-2. It is shown that ice sheets, changes in atmospheric compositions, vegetation cover, and reorganization of the ocean thermohaline circulation play important roles in glacial climate changes. Another example of this approach is the modelling of two major types of abrupt glacial climate changes: Dansgaard-Oeschger and Heinrich events. Our results corroborate some of the early proposed mechanisms, which relate abrupt climate changes to the internal instability of the ocean thermohaline circulation and ice sheets. At the same time, it is shown that realistic representation of the temporal evolution of the palaeoclimatic background is crucial to simulate observed features of the glacial abrupt climate changes. PMID- 14558900 TI - Evidence for rapid climate change in the Mesozoic-Palaeogene greenhouse world. AB - The best-documented example of rapid climate change that characterized the so called 'greenhouse world' took place at the time of the Palaeocene-Eocene boundary: introduction of isotopically light carbon into the ocean-atmosphere system, accompanied by global warming of 5-8 degrees C across a range of latitudes, took place over a few thousand years. Dissociation, release and oxidation of gas hydrates from continental-margin sites and the consequent rapid global warming from the input of greenhouses gases are generally credited with causing the abrupt negative excursions in carbon- and oxygen-isotope ratios. The isotopic anomalies, as recorded in foraminifera, propagated downwards from the shallowest levels of the ocean, implying that considerable quantities of methane survived upward transit through the water column to oxidize in the atmosphere. In the Mesozoic Era, a number of similar events have been recognized, of which those at the Triassic-Jurassic boundary, in the early Toarcian (Jurassic) and in the early Aptian (Cretaceous) currently carry the best documentation for dramatic rises in temperature. In these three examples, and in other less well-documented cases, the lack of a definitive time-scale for the intervals in question hinders calculation of the rate of environmental change. However, comparison with the Palaeocene-Eocene thermal maximum (PETM) suggests that these older examples could have been similarly rapid. In both the early Toarcian and early Aptian cases, the negative carbon-isotope excursion precedes global excess carbon burial across a range of marine environments, a phenomenon that defines these intervals as oceanic anoxic events (OAEs). Osmium-isotope ratios ((187)Os/(188)Os) for both the early Toarcian OAE and the PETM show an excursion to more radiogenic values, demonstrating an increase in weathering and erosion of continental crust consonant with elevated temperatures. The more highly buffered strontium-isotope system ((87)Sr/(86)Sr) also shows relatively more radiogenic signatures during the early Toarcian OAE, but the early Aptian and Cenomanian-Turonian OAEs show the reverse effect, implying that increased rates of sea-floor spreading and hydrothermal activity dominated over continental weathering in governing sea water chemistry. The Cretaceous climatic optimum (late Cenomanian to mid Turonian) also shows evidence for abrupt cooling episodes characterized by episodic invasion of boreal faunas into temperate and subtropical regions and changes in terrestrial vegetation; drawdown of CO(2) related to massive marine carbon burial (OAE) may be implicated here. The absence of a pronounced negative carbon-isotope excursion preceding the Cenomanian-Turonian OAE indicates that methane release is not necessarily connected to global deposition of marine organic carbon, but relative thermal maxima are common to all OAEs. 'Cold snaps' have also been identified from the Mesozoic record but their duration, causes and effects are poorly documented. PMID- 14558901 TI - Recent changes in the North Atlantic. AB - It has long been recognized that the Atlantic meridional overturning circulation (MOC) is potentially sensitive to greenhouse-gas and other climate forcing, and that changes in the MOC have the potential to cause abrupt climate change. However, the mechanisms remain poorly understood and our ability to detect these changes remains incomplete. Four main (interrelated) types of ocean change in particular are associated in the literature with greenhouse-gas forcing. These are: a slowing of MOC overturning rate; changes in northern seas which might effect a change in Atlantic overturning, including changes in the freshwater flux from the Arctic, and changes in the transport and/or hydrographic character of the northern overflows which ventilate the deep Atlantic; a change in the trans ocean gradients of steric height (both zonal and meridional) which might accompany a change in the MOC; and an intensification of the global water cycle. Though as yet we have no direct measure of the freshwater flux passing from the Arctic to the Atlantic either via the Canadian Arctic Archipelago or along the East Greenland Shelf, and no direct measure yet of the Atlantic overturning rate, we examine a wide range of time-series from the existing hydrographic record for oceanic evidence of the other anticipated responses. Large amplitude and sustained changes are found (or indicated by proxy) over the past three to four decades in the southward transport of fresh waters along the Labrador shelf and slope, in the hydrography of the deep dense overflows from Nordic seas, in the transport of the eastern overflow through Faroe Bank Channel, and in the global hydrologic cycle. Though the type and scale of changes in ocean salinity are consistent with an amplification of the water cycle, we find no convincing evidence of any significant, concerted slowdown in the Atlantic overturning circulation. PMID- 14558902 TI - Sea-ice switches and abrupt climate change. AB - We propose that past abrupt climate changes were probably a result of rapid and extensive variations in sea-ice cover. We explain why this seems a perhaps more likely explanation than a purely thermohaline circulation mechanism. We emphasize that because of the significant influence of sea ice on the climate system, it seems that high priority should be given to developing ways for reconstructing high-resolution (in space and time) sea-ice extent for past climate-change events. If proxy data can confirm that sea ice was indeed the major player in past abrupt climate-change events, it seems less likely that such dramatic abrupt changes will occur due to global warming, when extensive sea-ice cover will not be present. PMID- 14558903 TI - Atmospheric processes and observations. AB - The central role of the atmosphere in abrupt climate change is proposed and discussed. This discussion is given in the context of the poleward transport of energy in the climate system and of climate variability and change. A number of examples based on observational and model data are used to illustrate the ideas. PMID- 14558904 TI - Global warming and thermohaline circulation stability. AB - The Atlantic thermohaline circulation (THC) plays an important role in global climate. Theoretical and palaeoclimatic evidence points to the possibility of rapid changes in the strength of the THC, including a possible quasi-permanent shutdown. The climatic impacts of such a shutdown would be severe, including a cooling throughout the Northern Hemisphere, which in some regions is greater in magnitude than the changes expected from global warming in the next 50 years. Other climatic impacts would likely include a severe alteration of rainfall patterns in the tropics, the Indian subcontinent and Europe. Modelling the future behaviour of the THC focuses on two key questions. (i) Is a gradual weakening of the THC likely in response to global warming, and if so by how much? (ii) Are there thresholds beyond which rapid or irreversible changes in the THC are likely? Most projections of the response of the THC to increasing concentrations of greenhouse gases suggest a gradual weakening over the twenty-first century. However, there is a wide variation between different models over the size of the weakening. Rapid or irreversible THC shutdown is considered a low-probability (but high-impact) outcome; however, some climate models of intermediate complexity do show the possibility of such events. The question of the future of the THC is beset with conceptual, modelling and observational uncertainties, but some current and planned projects show promise to make substantial progress in tackling these uncertainties in future. PMID- 14558905 TI - The future of the carbon cycle: review, calcification response, ballast and feedback on atmospheric CO2. AB - The operation of the carbon cycle forms an important part of the processes relevant to future changes in atmospheric carbon dioxide. The balance of carbon between terrestrial and oceanic reservoirs is an important factor and here we focus in particular on the oceans. Future changes in the carbon cycle that may affect air-sea partitioning of CO(2) are difficult to quantify but the palaeoceanographic record and modern observational studies provide important evidence of what variations might occur. These include changes in surface nutrient use, the oceanic inventory of nutrients, and the elemental composition and rain-rate ratio of marine particles. Recent work has identified two inter linked processes of potential importance that we consider in some detail: the response of marine calcification to changes in surface water CO(2) and the association of particulate organic carbon with ballast minerals, in particular biogenic calcite. We review evidence from corals, coccolithophores and foraminifera, which suggests that the response of reduced calcification provides a negative feedback on rising atmospheric CO(2). We then use a box model to demonstrate how the calcification response may affect the organic carbon rain rate through the ballast effect. The ballast effect on export fluxes of organic and inorganic carbon acts to counteract the negative calcification response to increased CO(2). Thus, two oceanic buffers exert a significant control on ocean atmosphere carbonate chemistry: the thermodynamic CO(2) buffer; and the ballast/calcification buffer. Just how tightly coupled the rain-rate ratio of CaCO(3)/C(org) is to fluxes of ballast minerals is an important question for future research. PMID- 14558906 TI - Abrupt climate change: can society cope? AB - Consideration of abrupt climate change has generally been incorporated neither in analyses of climate-change impacts nor in the design of climate adaptation strategies. Yet the possibility of abrupt climate change triggered by human perturbation of the climate system is used to support the position of both those who urge stronger and earlier mitigative action than is currently being contemplated and those who argue that the unknowns in the Earth system are too large to justify such early action. This paper explores the question of abrupt climate change in terms of its potential implications for society, focusing on the UK and northwest Europe in particular. The nature of abrupt climate change and the different ways in which it has been defined and perceived are examined. Using the example of the collapse of the thermohaline circulation (THC), the suggested implications for society of abrupt climate change are reviewed; previous work has been largely speculative and has generally considered the implications only from economic and ecological perspectives. Some observations about the implications from a more social and behavioural science perspective are made. If abrupt climate change simply implies changes in the occurrence or intensity of extreme weather events, or an accelerated unidirectional change in climate, the design of adaptation to climate change can proceed within the existing paradigm, with appropriate adjustments. Limits to adaptation in some sectors or regions may be reached, and the costs of appropriate adaptive behaviour may be large, but strategy can develop on the basis of a predicted long term unidirectional change in climate. It would be more challenging, however, if abrupt climate change implied a directional change in climate, as, for example, may well occur in northwest Europe following a collapse of the THC. There are two fundamental problems for society associated with such an outcome: first, the future changes in climate currently being anticipated and prepared for may reverse and, second, the probability of such a scenario occurring remains fundamentally unknown. The implications of both problems for climate policy and for decision making have not been researched. It is premature to argue therefore that abrupt climate change - in the sense referred to here - imposes unacceptable costs on society or the world economy, represents a catastrophic impact of climate change or constitutes a dangerous change in climate that should be avoided at all reasonable cost. We conclude by examining the implications of this contention for future research and policy formation. PMID- 14558907 TI - Ecological impacts of atmospheric CO2 enrichment on terrestrial ecosystems. AB - Global change has many facets, of which land use and the change of atmospheric chemistry are unquestioned primary agents, which induce a suite of secondary effects, including climatic changes. The largest single contribution to the compositional change of the atmosphere, CO(2) enrichment, has (besides its influence on climate) immediate and direct effects on plants. Quantitatively, CO(2) is the plant 'food' number one, and the rate of photosynthetic CO(2) uptake by leaves is not yet CO(2)-saturated. This abrupt change of the biosphere's diet does and will affect all aspects of life, including our food. However, the plant and ecosystem responses are more subtle than had been assumed from the results of responses of isolated, well-fertilized and well-watered plants in greenhouses during the early days of CO(2)-enrichment research. In this article, I discuss potential responses of complex natural grassland and diverse forests, and address three key themes: CO(2) and nutrients; CO(2) and water; CO(2) and plant-animal interactions. Examples from a suite of climatic regions emphasize that the most important ecosystem level responses to elevated CO(2) will be introduced by differential responses of species. Atmospheric CO(2) enrichment is a biodiversity issue. Classical physiological baseline responses of leaves to elevated CO(2) can be overrun by biodiversity effects to such an extent that some of the traditional predictions may even become reversed. For instance, biodiversity effects may cause humid tropical forests (those which avoid destruction) to become more dynamic and store less, rather than more, carbon as CO(2) enrichment continues. The abundance of certain life forms and species and their lifespans exert major controls over the half-life of carbon stored in forest biomass, and there is evidence that elevated CO(2) can affect these controls and most likely does so already. Also, long-term hydrological consequences of atmospheric CO(2) enrichment will be driven by biodiversity effects, given that some, but not all, species reduce their rate of water loss when exposed to a CO(2)-rich atmosphere. Such insights call for much more realistic experimental conditions and larger scale test units, which permit biotic interactions across taxa and trophic levels to occur while simulating our CO(2) future. The evidence currently available suggests that ecosystem processes reflect the composition of their biological inventory and this will be affected by a shift in carbon supply. PMID- 14558908 TI - The economics of abrupt climate change. AB - The US National Research Council defines abrupt climate change as a change of state that is sufficiently rapid and sufficiently widespread in its effects that economies are unprepared or incapable of adapting. This may be too restrictive a definition, but abrupt climate change does have implications for the choice between the main response options: mitigation (which reduces the risks of climate change) and adaptation (which reduces the costs of climate change). The paper argues that by (i) increasing the costs of change and the potential growth of consumption, and (ii) reducing the time to change, abrupt climate change favours mitigation over adaptation. Furthermore, because the implications of change are fundamentally uncertain and potentially very high, it favours a precautionary approach in which mitigation buys time for learning. Adaptation-oriented decision tools, such as scenario planning, are inappropriate in these circumstances. Hence learning implies the use of probabilistic models that include socioeconomic feedbacks. PMID- 14558909 TI - Rapid climate change: scientific challenges and the new NERC programme. AB - In this paper the scientific challenges of observing, modelling, understanding and predicting rapid changes in climate are discussed, with a specific focus on the role of the Atlantic thermohaline circulation. The palaeo and present-day observational and modelling studies being carried out to meet these challenges, under the aegis of a new NERC Rapid Climate Change thematic programme (RAPID), are outlined. In particular, the paper describes the work being done to monitor changes in the meridional overturning circulation of the North Atlantic. The paper concludes with some speculative comments about potential mechanisms for rapid changes. PMID- 14558911 TI - Accuracy-based learning classifier systems: models, analysis and applications to classification tasks. AB - Recently, Learning Classifier Systems (LCS) and particularly XCS have arisen as promising methods for classification tasks and data mining. This paper investigates two models of accuracy-based learning classifier systems on different types of classification problems. Departing from XCS, we analyze the evolution of a complete action map as a knowledge representation. We propose an alternative, UCS, which evolves a best action map more efficiently. We also investigate how the fitness pressure guides the search towards accurate classifiers. While XCS bases fitness on a reinforcement learning scheme, UCS defines fitness from a supervised learning scheme. We find significant differences in how the fitness pressure leads towards accuracy, and suggest the use of a supervised approach specially for multi-class problems and problems with unbalanced classes. We also investigate the complexity factors which arise in each type of accuracy-based LCS. We provide a model on the learning complexity of LCS which is based on the representative examples given to the system. The results and observations are also extended to a set of real world classification problems, where accuracy-based LCS are shown to perform competitively with respect to other learning algorithms. The work presents an extended analysis of accuracy-based LCS, gives insight into the understanding of the LCS dynamics, and suggests open issues for further improvement of LCS on classification tasks. PMID- 14558912 TI - Analysis and improvement of fitness exploitation in XCS: bounding models, tournament selection, and bilateral accuracy. AB - The evolutionary learning mechanism in XCS strongly depends on its accuracy-based fitness approach. The approach is meant to result in an evolutionary drive from classifiers of low accuracy to those of high accuracy. Since, given inaccuracy, lower specificity often corresponds to lower accuracy, fitness pressure most often also results in a pressure towards higher specificity. Moreover, fitness pressure should cause the evolutionary process to be innovative in that it combines low-order building blocks of lower accurate classifiers, to higher-order building blocks with higher accuracy. This paper investigates how, when, and where accuracy-based fitness results in successful rule evolution in XCS. Along the way, a weakness in the current proportionate selection method in XCS is identified. Several problem bounds are derived that need to be obeyed to enable proper evolutionary pressure. Moreover, a fitness dilemma is identified that causes accuracy-based fitness to be misleading. Improvements are introduced to XCS to make fitness pressure more robust and overcome the fitness dilemma. Specifically, (1) tournament selection results in a much better fitness-bias exploitation, and (2) bilateral accuracy prevents the fitness dilemma. While the improvements stand for themselves, we believe they also contribute to the ultimate goal of an evolutionary learning system that is able to solve decomposable machine-learning problems quickly, accurately,and reliably. The paper also contributes to the further understanding of XCS in general and the fitness approach in XCS in particular. PMID- 14558913 TI - Bounding the effect of noise in multiobjective learning classifier systems. AB - This paper analyzes the impact of using noisy data sets in Pittsburgh-style learning classifier systems. This study was done using a particular kind of learning classifier system based on multiobjective selection. Our goal was to characterize the behavior of this kind of algorithms when dealing with noisy domains. For this reason, we developed a theoretical model for predicting the minimal achievable error in noisy domains. Combining this theoretical model for crisp learners with graphical representations of the evolved hypotheses through multiobjective techniques, we are able to bound the behavior of a learning classifier system. This kind of modeling lets us identify relevant characteristics of the evolved hypotheses, such as overfitting conditions that lead to hypotheses that poorly generalize the concept to be learned. PMID- 14558914 TI - For real! XCS with continuous-valued inputs. AB - Many real-world problems are not conveniently expressed using the ternary representation typically used by Learning Classifier Systems and for such problems an interval-based representation is preferable. We analyse two interval based representations recently proposed for XCS, together with their associated operators and find evidence of considerable representational and operator bias. We propose a new interval-based representation that is more straightforward than the previous ones and analyse its bias. The representations presented and their analysis are also applicable to other Learning Classifier System architectures. We discuss limitations of the real multiplexer problem, a benchmark problem used for Learning Classifier Systems that have a continuous-valued representation, and propose a new test problem, the checkerboard problem, that matches many classes of real-world problem more closely than the real multiplexer. Representations and operators are compared using both the real multiplexer and checkerboard problems and we find that representational, operator and sampling bias all affect the performance of XCS in continuous-valued environments. PMID- 14558915 TI - Believe it or not, Europe can do some things well. PMID- 14558917 TI - Thyrotropin effects on ultraviolet radiation-dependent apoptosis in FRTL-5 cells. AB - Apoptosis plays an important role within the endocrine system, particularly in the thyroid gland, although little is known about its regulation in normal thyroids. Because thyrotropin (TSH) regulates many thyroid-specific functions and cell proliferation, we investigated whether TSH can influence such mechanisms. To induce apoptosis we used UV-C radiation. The FRTL-5 rat thyroid cell strain, a cloned strain of differentiated and untransformed cells that reproduces many of the characteristics of the normal thyroid was chosen for this study. The FRTL-5 cells are a particularly suitable model because they actively proliferate when cultured in the presence of TSH (6H medium), while in TSH-free medium (5H medium) cells remain in a physiologic quiescent state for a long period of time. FRTL-5 cells in both culture conditions were irradiated with UV-C radiation (254 nm wavelength). At 48 hours after radiation, 6H cultured cells showed the characteristic signs of apoptosis. However, 5H cultured cells did not present macroscopic signs of damage, DNA fragmentation, or detectable apoptosis. Furthermore, the expression of 23 apoptosis-related genes was compared. Results indicate that Bcl2 and caspase-2 expression is enhanced, while bax, GADD45 and mdm-2 expression is reduced in irradiated cells. These data confirm that TSH plays a major role in regulating UV-induced apoptosis in FRTL-5 cells. PMID- 14558918 TI - Interpretation of sex hormone-binding globulin levels in thyroid disorders. AB - Sex hormone-binding globulin (SHBG) levels were followed in three groups of female subjects to evaluate interrelationships between its levels and parameters characterizing thyroid function. In the first part of the study the data from 201 patients with thyroid and other endocrine dysfunctions were grouped according to SHBG or basic laboratory parameters of thyroid function (thyrotropin, free thyroxine). Particular attention was paid to the presence of antithyroid antibodies (against thyroglobulin or thyroid peroxidase). Analysis of covariance revealed that SHBG changes were significant only in hyperthyroidism, and were not influenced by the presence of antibodies. Age was a minor factor influencing SHBG levels, in contrast to thyroid status. In a well-defined group of 16 females with severe hypothyroidism after total thyroidectomy because of thyroid cancer the low SHBG levels increased significantly to physiologic values after reaching normal thyroid function, irrespective of contraceptive use. In the final part of the study SHBG levels were correlated with the basic laboratory data, reflecting thyroid function in a sample of normal female population (129 subjects) screened for iodine deficiency in one region of the Czech Republic. After adjustment for age, the only significant positive correlation was between SHBG and free triiodothyronine. PMID- 14558919 TI - Sibling recurrence risk in autoimmune thyroid disease. AB - There is abundant evidence for a genetic influence on the development of autoimmune thyroid diseases (AITD). One measure of the magnitude of genetic contribution to the development of a disease is the sibling risk ratio (lambda(s)). Recent accurate prevalence data for hypothyroidism and hyperthyroidism in the United States reported from the National Health and Nutrition Examination Survey III (NHANES III) study have now allowed us to compute the sibling recurrence risk for AITD. Patients were recruited from our endocrine clinic on the basis of having AITD. The inclusion of patients in this study was unambiguously single ascertainment. We studied 155 patients (131 with Graves' disease [GD] and 24 with Hashimoto's thyroiditis [HT]) who had reliable information on the presence or absence of AITD in siblings. Nine probands had siblings with GD and 13 probands had siblings with HT. Using the prevalence rates from NHANES III for clinical hyperthyroidism and hypothyroidism, the calculated lambda(s) was 16.9 for AITD, 11.6 for GD, and 28.0 for HT. These results confirm the significant contribution of genetic factors to the development of AITD. PMID- 14558920 TI - Galectin-3 and oncofetal-fibronectin expression in thyroid neoplasia as assessed by reverse transcription-polymerase chain reaction and immunochemistry in cytologic and pathologic specimens. AB - Oncofetal fibronectin (onfFN) and galectin-3 (Gale-3) have been proposed as possible tools for the preoperative diagnosis of thyroid carcinomas, based on the finding that the expression of both onfFN and Gale-3 are significantly increased in papillary and anaplastic carcinomas, compared to normal thyroid tissues and follicular adenomas. In this study we analyzed the expression of these markers by immunochemical and molecular analysis of benign and malignant thyroid tumors. Sixty-five thyroid nodules, consisting of 20 follicular adenomas (FAs) and 45 papillary thyroid carcinomas (PTCs) at final histology were examined. At the molecular level, among the 45 PTCs, 44 (97.8%) showed a variable level of onfFN mRNA, while 8 of the 20 (40%) adenomas expressed the same marker. Similar results have been found analyzing Gale-3 expression: 97.8% of PTC and 55% of FAs were positive for this marker. Immunohistochemistry (IHC) for Gale-3 was positive in 42 of 45 (93.3%) PTC tissues. Staining was invariably confined to the cytoplasm, with a homogeneous distribution in the large majority of the neoplastic cells. The 3 negative cases (6.7%) were represented by 2 classic variants of PTC and 1 follicular variant of PTC. Eighteen of the 20 (90.0%) adenomas stained negative for Gale-3. A significant association was found between positive staining and malignant phenotype (p < 0.0001). Gale-3 protein expression was also performed on samples obtained by ex vivo fine-needle aspiration (FNA) in 35 PTCs by immunocytochemistry (ICC). Immunoreactivity was present in 32 (91.0%) and negative in 3 (8.8%) cases. With the exception of 1 case (negative by ICC and positive by IHC), ICC and IHC were fully concordant. In conclusion, our results indicate that a search for Gale-3 protein overexpression by IIC or ICC, but not by reverse transcription-polymerase chain reaction (RT-PCR), may yield an additional marker of malignant potential of thyroid nodular lesions, and may be a useful adjunct to the currently available diagnostic tools for the preoperative diagnosis of malignant thyroid tumors. PMID- 14558921 TI - An outline of inherited disorders of the thyroid hormone generating system. AB - To date, various genetic defects impairing the biosynthesis of thyroid hormone have been identified. These congenital heterogeneous disorders result from mutations of genes involved in many steps of thyroid hormone synthesis, storage, secretion, delivery, or utilization. In contrast to thyroid dyshormonogenesis, the elucidation of the underlying etiology of most cases of thyroid dysgenesis is much less understood. It is suggested that genetic factors might play a role in some cases of thyroid dysgenesis and the best candidate genes involved are those encoding transcription factors known to play a role in the embryonic development of the thyroid gland. Moreover, discordance for thyroid dysgenesis is the rule for monozygotic twins as recently reported and this may result from epigenetic phenomena, early somatic mutations, or postzygotic events. In the final part of this review the molecular defects involved in proteins that transport thyroid hormone in the circulation are described: thyroxine-binding globulin (TBG), transtiretin and albumin, that may be associated with altered thyroid function tests and other pathologic conditions such as amyloidotic polyneuropathy. PMID- 14558922 TI - The prevalence of undiagnosed thyroid disorders in a previously iodine-deficient area. AB - OBJECTIVE: The aim of the present study was to analyze the current status of morphologic and functional thyroid abnormalities in a previously iodine-deficient area. METHODS: The population based Study of Health in Pomerania (SHIP) comprised 4310 participants, aged 20-79 years. Thyroid function (thyrotropin [TSH] free triiodothyronine [FT(3)], and free thyroxine [FT(4)]) and serum autoantibodies to thyroperoxidase (TPOAb) were evaluated from blood samples. Thyroid structure and size were measured by ultrasound. Data from 3941 participants with no known thyroid disorders were analyzed. RESULTS: The median iodine urine excretion was 12.4 microg/dL. The rate of decreased serum TSH levels (<0.3 mIU/L) was 11.3%; 2.2% of participants had suppressed serum TSH levels (<0.1 mIU/L). The prevalence of subclinical hyperthyroidism was 1.8%, the prevalence of overt hyperthyroidism 0.4%. Elevated TSH levels were found in 1.2% of individuals. Subclinical hypothyroidism was observed in 0.5%, overt hypothyroidism in 0.7% of the sample. Elevated TPOAb were detected in 7% of subjects, 4.1% of participants had TPOAb greater than 200 IU/mL. The prevalence of goiter was 35.9%. An inhomogeneous echo pattern was detected in 35.2% and nodules in 20.2% of participants. Diffuse autoimmune thyroiditis was diagnosed in 47 subjects (1.2%). CONCLUSION: There are a number of thyroid disorders in this previously iodine-deficient region. Further studies are required to investigate the change of thyroid disorders during iodine supplementation programs. PMID- 14558923 TI - Association of seasonal allergic rhinitis is high in Graves' disease and low in painless thyroiditis. AB - Hashimoto's thyroiditis is thought to be a T-helper cell type 1 (TH1)-dependent disease, but it is not clear whether Graves' disease is T-helper cell type 2 (TH2)-predominant or not. TH1-predominant diseases are infrequently and TH2 predominant diseases are frequently associated with allergic diseases. We examined the prevalence of seasonal allergic rhinitis to Japanese cedar pollen, a typical TH2-associated disease, in patients with Graves' disease (n = 126), painless thyroiditis (n = 46) and Hashimoto's thyroiditis (n = 88), and compared them to healthy controls (n = 766). Gender and age distribution were not different among patient groups and healthy controls, except for the higher age of patients with Hashimoto's thyroiditis. The prevalence of seasonal allergic rhinitis was significantly high in patients with Graves' disease (42.9%, p < 0.05) and low in patients with painless thyroiditis (13.0%, p < 0.01) but was not different in patients with Hashimoto's thyroiditis (26.1%) compared to that of healthy controls (32.6%). When patients with painless thyroiditis were included in Hashimoto's thyroiditis group, the prevalence of seasonal allergic rhinitis was 21.6% and significantly different from that of healthy controls (p < 0.05). These data indicate that Graves' disease is TH2 predominant and painless thyroiditis is a TH1-predominant disease. Our findings suggest that the shift from TH2 toward TH1 immunogenesis may be important for achieving earlier remission of Graves' disease. PMID- 14558924 TI - Possible induction of Graves' disease and painless thyroiditis by gonadotropin releasing hormone analogues. AB - Prolonged administration of gonadotropin-releasing hormone (GnRH) analogues induce a decrease in serum estrogen level, which may aggravate subclinical or mild autoimmune thyroid disease. Two patients developed Graves' thyrotoxicosis in association with an increase in anti-thyrotropin (TSH) receptor antibody activities at 4 months after initiation of buserelin acetate. GnRH analogue therapy was discontinued at the time of diagnosis but it took more than 2 years of methimazole therapy to obtain remission of Graves' disease. Another patient developed painless thyroiditis in association with an increase in antithyroid microsomal antibodies at 4 months after initiation of leuprolide acetate. These results indicate that GnRH analogues possibly induce clinical onset of Graves' thyrotoxicosis or destruction-induced thyrotoxicosis. Clinicians should be aware of this phenomenon. All patients who are to receive GnRH analogue therapy should be examined for antithyroid antibodies and family history of autoimmune thyroid disease, and should be followed accordingly. PMID- 14558926 TI - Severe Graves' ophthalmopathy after retrobulbar anesthesia for cataract extraction in a patient with mild stable thyroid eye disease. AB - It has been hypothesized that the distinct anatomic localization of the Graves' triad may be partially explained by pressure and trauma. While there are reports of local trauma clearly contributing to the pathogenesis of pretibial myxedema, direct evidence for a similar mechanism in Graves' ophthalmopathy (GO) has been lacking. We describe a 65-year-old male patient with stable mild Graves' ophthalmopathy of 24 years' duration in whom a retrobulbar block was administered prior to cataract removal. Three weeks after the procedure, he complained of rapidly progressive bilateral diplopia. In 6 months, there was moderate exophthalmos, exposure keratitis, almost complete ophthalmoplegia, and decreasing visual acuity requiring surgical decompression. Postdecompression, inflammatory signs and vision improved but there was complete ophthalmoplegia. The eye signs remained unchanged for the next 4 months but there was exacerbation of the disease within a week of receiving radioiodine despite concomitant steroid administration. Orbital irradiation was finally administered with rapid improvement in extraocular eye muscle function. We hypothesize that local inflammatory and immune responses stimulated by trauma and/or pressure in the retrobulbar compartment, triggered the development of severe ophthalmopathy in this patient. Thyroid-stimulating immunoglobulin (TSI) levels remained markedly elevated despite the clinical improvement suggesting that the beneficial effects of radiotherapy in this case were not mediated by suppressing TSI production. PMID- 14558925 TI - Calcitonin precursor levels in human medullary thyroid carcinoma. AB - DESIGN: The hormonal serum marker for the presence and course of patients with medullary thyroid cancer (MTC) is the mature calcitonin (CT) peptide. Other CALC 1 gene products such as the 116-amino acid polypeptide prohormone, procalcitonin, as well as its component calcitonin precursors (CTpr) may also be increased in their sera. We performed a study to evaluate the clinical utility of serum levels CTpr in these patients. METHODS: Twenty-one patients with MTC (9 males, 12 females; 23-76 years of age) were evaluated. The diagnosis was confirmed by histologic examination, except for 2 (a proven RET mutation plus an abnormal pentagastrin-stimulated CT level). Nine patients had postoperative hypercalcitoninemia and 3 of these died. The specific assay for mature CT was a commercial immunoradiometric assay (hCT-IRMA); the immunoluminometric assay for CTpr (B.R.A.H.M.S Diagnostica, Berlin, Germany) detects intact procalcitonin and the free CT:CT carboxypeptide-1. RESULTS: All patients had detectable serum CTpr. These levels considerably exceeded those of mature CT, averaging 7.6-fold greater. CTpr levels correlated positively with mature CT (r = 0.61; p < 0.001). After pentagastrin administration, there was a parallelism of response between the two assays. Whenever there were known metastases, CTpr increased markedly. CONCLUSION: This study demonstrates the universal presence of CTpr in the blood of patients with MTC. The measurement of these peptides may offer a new dimension to the clinical evaluation of this malignancy. PMID- 14558927 TI - Near-lethal respiratory failure after recombinant human thyroid-stimulating hormone use in a patient with metastatic thyroid carcinoma. AB - A patient with widely metastatic differentiated thyroid cancer who had been heavily pretreated with (131)I was given recombinant human thyroid stimulating hormone (rhTSH) prior to (131)I treatment. Clinical and physical data from both this case and the literature suggest that the recombinant hormone, not the (131)I, may have caused a significant portion of the tumor swelling, which in turn was the most likely cause of the patient's symptoms. The potential effect of (131)I-induced tumor swelling and direct radiation effect on the lung is also analyzed. We review the potential hazards associated with rhTSH in patients with metastasis and propose means of minimizing this risk. PMID- 14558928 TI - Pituitary hyperplasia during primary hypothyroidism. PMID- 14558929 TI - Thyroglobulin-positive, radioiodine-negative thyroid cancer. PMID- 14558930 TI - A rapid high-performance liquid chromatographic method for the simultaneous quantitation of aspirin, salicylic acid, and caffeine in effervescent tablets. AB - A rapid reversed-phase high-performance liquid chromatographic procedure is developed and validated for the simultaneous quantitation of aspirin, salicylic acid, and caffeine extracted from an effervescent tablet. The method uses a Hypersil C18 column (5 micro m, 15 cm x 4.6 mm) for an isocratic elution in a water-methanol-acetic acid mobile phase at a wavelength of 275 nm. The tablets' buffering effects and acid neutralizing capacity require an extraction solvent of methanol-formic acid. The range of linearity for aspirin is 0.5-1.25 mg/mL, caffeine 0.065-0.195 mg/mL, and salicylic acid 0.4-6.0% of aspirin. The overall recovery is 100.2%, 100.7%, and 99.2% for aspirin, caffeine, and salicylic acid, respectively. Under the conditions of the method, aspirin, caffeine, and salicylic acid are adequately resolved with proper peak symmetry in less than 7 min. PMID- 14558931 TI - Online hydrogen-deuterium exchange and a tandem-quadrupole time-of-flight mass spectrometer coupled with liquid chromatography for metabolite identification in drug metabolism. AB - A method for metabolite identification in drug discovery and development utilizing online hydrogen-deuterium (H-D) exchange and a tandem-quadrupole time of-flight mass spectrometer coupled with liquid chromatography has been developed for the first time. For purposes of evaluating this method, nimodipine is incubated with rat, dog, monkey, and human hepatic microsomes over a 60-min period. This procedure involves the replacement of H(2)O with D(2)O in the mobile phase. The equilibration time required to change (switch) from H(2)O in the mobile phase into D(2)O is approximately 10 min. There is no significant difference between the retention times in H(2)O or D(2)O mobile phase. The results show that the separations of nimodipine and its metabolites and the subsequent H-D exchanges were achieved online. This leads to the identification of nimodipine metabolites formed in vitro. Five metabolites of nimodipine are identified and characterized. These metabolites are formed by dehydrogenation, demethylation of methoxy group, cleavage of the ester groups by hydrolysis or oxidation, and hydroxylation of methyl groups. The combination of the H-D exchange process and the assay system is found to be a powerful tool for the study of metabolite identification. The method appears to be highly desirable for identification of metabolites produced by dehydrogenation, oxidation, and dealkylation, which are discussed herein. This method also greatly enhances throughput, which in turn facilitates the ability to rapidly provide metabolite elucidation. PMID- 14558932 TI - Liposomal benznidazole: a high-performance liquid chromatographic determination for biodistribution studies. AB - In this work, an isocratic high-performance liquid chromatographic method for quantitation of liposomal benznidazole (BNZ) in biological tissues is presented. The method comprises protein precipitation together with an efficient extraction of bulk or liposomal BNZ with acetonitrile-dimethylsulfoxide (1:1, v/v) at a 2:1 (extraction solvent-tissue matrix, v/v or /vw) ratio; the process is completed by a final precipitation with trichloroacetic acid. The resultant supernatants are assayed chromatographically using a Kromasil C18 (25- x 0.4-cm i.d., 100 A, 5- microm particle size), with an isocratic mobil phase consisting of acetonitrile water (40:60, v/v), a flow rate of 0.9 mL/min, and detected at 324 nm. Bulk BNZ is used as a reference standard for the analysis of samples containing liposomal BNZ. The assay is linear over a concentration range of 0.75 (the lowest quantity of analyte determined with precision and accuracy of >or= 20%) to 25 microg/mL-g in all liquid and solid matrices. Within-day precision is better than 6.4% in plasma and 8.6% in liver, the same for the two assayed concentrations. Between day precision is 5.4% and 12.3% in plasma and 9% and 6.9% in liver for the two assayed concentrations, respectively. The absolute recoveries range between 70% and 97%. Therefore, the method is accurate and precise to be employed for detection of minor quantities of liposomal BNZ in biological tissues. PMID- 14558933 TI - Determination of 1,4-dioxane impurity levels in Triton X-100 raw material by gas chromatography with mass spectrometric detection. AB - Triton X-100 (octoxynol 9) is a commercially available surfactant used as a solvent detergent in numerous pharmaceutical applications including virus inactivation. A byproduct formed during its synthesis is 1,4-dioxane, the cyclic dimer of ethylene oxide and a possible carcinogen to humans. The United States Pharmacopoeia (USP) contains a labor-intensive 1,4-dioxane test for Triton X-100. The method couples vacuum distillation to extract the 1,4-dioxane from the Triton X-100 matrix followed by gas chromatography (GC) using a packed column with flame ionization detection. In order to provide a more automated and specific test methodology, a headspace GC-mass spectrometry (MS) method has been developed for this application. Analyte quantitation is accomplished by the method of standard additions. The automated sample preparation, coupled with the specificity inherent in high-efficiency capillary column separations together with single-ion MS detection, results in an assay that is more efficient, accurate, and precise than the USP procedure. Performance characteristics of the headspace GC-MS method are contrasted with those characteristics of the USP methodology. PMID- 14558934 TI - Development and validation of a liquid chromatographic method for monitoring of process-related synthetic organic impurities of profenofos in technical products. AB - A simple and rapid reversed-phase high-performance liquid chromatographic method for the monitoring of process-related synthetic organic impurities of profenofos (PFS) is developed. Impurities are separated and determined on a reversed-phase Hypersil C(18) column using gradient elution of 50 mM ammonium formate buffer acetonitrile as a mobile phase and detection at 230 nm at ambient temperature. The method is validated with respect to accuracy, precision, linearity, and limits of detection and quantitation. The method is found to be suitable not only for monitoring the reactions involved in the process development of PFS, but also quality assurance, as it can detect impurities at the level of 1.5 x 10(-8) g. PMID- 14558935 TI - Investigation of liquid and gas chromatography techniques for separation of diastereomers of beta-(alpha-methylbenzyl) amino isobutyric acid. AB - Cryptophycins are macrolides investigated as potential anticancer agents. These large cyclic molecules are generated via a convergent process, utilizing the coupling of several smaller fragments synthesized individually. During early synthetic development of the beta-amino acid fragment C, analytical methods are necessary for the characterization of products resulting from the various routes being studied. One route being evaluated produces (RR) and (RS) diastereomers of beta-(alpha-methylbenzyl) amino isobutyric acid as intermediates. To measure diastereomeric excess (%de), assay conditions using high-performance liquid chromatography (HPLC) and capillary gas chromatographic (GC) techniques are explored. Derivatization methods using trifluoroacetyl- and silyl-derivatives are investigated for use with capillary GC. The results of the GC investigations are found to be only partially successful. Ion-pair HPLC is determined to be the optimal technique, utilizing pentanesulfonic acid as the counter ion to the amine group of beta-(alpha-methylbenzyl) amino isobutyric acid. PMID- 14558936 TI - Characterization of phthalides in Ligusticum chuanxiong by liquid chromatographic atmospheric pressure chemical ionization-mass spectrometry. AB - High-performance liquid chromatography (HPLC) with diode-array detection interfaced to atmospheric pressure chemical ionization (APCI)-mass spectrometry (MS) is applied to analyze phthalides from Chuanxiong (the rhizome of Ligusticum chuanxiong). This herb material, containing plenty of phthalide compositions, is selected as the analytical target in this paper for its hematological activity. Some of the phthalides are not stable and are difficult to analyze by gas chromatography-MS. Under optimized LC-MS-MS conditions, six phthalides in the methanol extract of Chuanxiong are unambiguously identified, and characteristic fragments are obtained using homemade reference standards. Ten other phthalides in the extract are confirmed by means of LC-APCI-MS with positive-negative ion mode and collision-induced dissociation in combination with UV spectrophotometry. The results show that LC-MS-MS is a method of choice for fast detection and detailed structural analysis of such mixtures in the crude extract of Chuanxiong. PMID- 14558937 TI - Simultaneous analyses of neutral carbohydrates and amino sugars in freshwaters with HPLC-PAD. AB - In this study, we determine concentrations of neutral and amino sugars and a sugar alcohol in freshwaters using high-performance liquid chromatography and pulsed amperometric detection with a single isocratic analysis. Coeluting arabinose, galactosamine, and mannosamine are separated with a mobile phase of 22.8 mM NaOH-KOH at a temperature of 17 degrees C. The resolutions are 0.73 and 0.64, respectively. The method separates closely eluting glucose-mannose and mannose-xylose peaks with resolutions of 0.85 and 0.71. Other sugars, fucose, rhamnose, galactose, fructose, ribose, glucosamine, and mannitol are resolved completely. Arabinose and galactosamine are measured in stream, ground, and soil waters that contain dissolved total saccharide (DTS) concentrations of 527 to 1555 nM. Failure to distinguish galactosamine from arabinose in those samples results in a 53-82% overestimation of arabinose concentrations and a 1.8-6.5% overestimation of DTS concentrations. The near unity of glucosamine and galactosamine concentrations in stream water samples allows us to suggest a correction factor for historical samples that had been analyzed without resolving galactosamine and arabinose. PMID- 14558938 TI - Measurement uncertainty of liquid chromatographic analyses visualized by Ishikawa diagrams. AB - Ishikawa, or cause-and-effect diagrams, help to visualize the parameters that influence a chromatographic analysis. Therefore, they facilitate the set up of the uncertainty budget of the analysis, which can then be expressed in mathematical form. If the uncertainty is calculated as the Gaussian sum of all uncertainty parameters, it is necessary to quantitate them all, a task that is usually not practical. The other possible approach is to use the intermediate precision as a base for the uncertainty calculation. In this case, it is at least necessary to consider the uncertainty of the purity of the reference material in addition to the precision data. The Ishikawa diagram is then very simple, and so is the uncertainty calculation. This advantage is given by the loss of information about the parameters that influence the measurement uncertainty. PMID- 14558939 TI - A simple and reliable semipreparative high-performance liquid chromatography technique for the isolation of marker-grade hyperforin from Hypericum perforatum L extract. AB - The present work describes isolation of bioactive lipophilic constituent [namely, hyperforin from St. John's wort (Hypericum perforatum L.)], of approximately 98% purity by semipreparative high-performance liquid chromatography (LC). The extraction, isolation, and analysis of the collected compound is performed without the use of antioxidants and inert gas atmospheres at all the stages. Hyperforin, separated isocratically on a 12microm semiprep column, is obtained in high purity, lyophilized after the removal of the organic phase, and preserved at a low temperature. The purity of the collected marker compound is estimated by the use of LC-mass spectrometry and spectroscopic techniques. PMID- 14558940 TI - [Construction of PTTGas antisense expression vector and its inhibitory effects on human ovarian carcinoma cell line SK-OV-3]. AB - BACKGROUND & OBJECTIVE: Pituitary tumor transforming gene (PTTG) is a new proto oncogene and shows multiple actions of promoting tumorigenesis and metastasis. Most researches mainly focus on the problems of PTTG expression in different tumor tissues and its relative regulatory mechanisms, but no research of exploring the possibility of antisense blocking of PTTG gene has been reported by now. In present study, the authors constructed eukaryotic expression vector expressing full-length anti-sense PTTG mRNA and observed its blocking effect on the potential invasion of human ovarian carcinoma cell line SK-OV-3. METHODS: PCR primers containing designed enzyme cut sites were used for cloning full-length PTTG gene fragment, and the resulting PCR product was inserted into the eukaryotic vector pcDNA3.1 in the antisense direction. The recombinant vector was then transfected into SK-OV-3 by lipofectamine. The positive cell clone was screened by G418,PTTG,and bFGF at protein level expression were detected by Western blot analysis. The changes of cell proliferation were analyzed by MTT method. The biological behavior change of transfection positive cells was observed by colony formation in soft agar assay. RESULTS: SK-OV-3 clones stably expressing full-length recombinant pcDNA3.1- PTTGas were obtained.The expression of PTTG and bFGF proteins in transfected cells were decreased by 61.5% and 52.3% respectively as compared with non-transfected ones. The cell proliferation was accelerated in transfected cells. The number of colony formation is reduced significantly in transfected cells (2.4+/-0.8) as compared with non-transfected and empty vector transfected cells (23.3+/-5.7 and 21.5+/-7.9, respectively, P< 0.01). CONCLUSION: The recombinant vector pcDNA3.1-PTTGas is a novel tool and brings us a new possibility of anti-sense gene therapy targeted at PTTG in human carcinoma. PMID- 14558941 TI - [Cloning and expression analysis of lung carcinoma related gene HLCDG1]. AB - BACKGROUND & OBJECTIVE: High frequent loss of hetero- zygosity (LOH) of 3p, 5q, 6q, 9p, 10q, 11p, 13q, 17p, and 19p in lung carcinoma was detected by comparative genomic hybridization (CGH) and genomic-wide scan for analysis of genetic alteration with microsatellite allelotying. It was indicated that there might be some other unknown tumor susceptible genes or suppressor genes likely to be involved in lung carcinoma development and progression. The aim of this study was to clone the full-length cDNA of LXDD1,an expressed sequence tag(EST) isolated by mRNA differential display, which is significantly down-regulated in lung carcinoma and represents a novel gene. METHODS: Differential expression of LXDD1 in lung carcinoma was confirmed by Northern blot analysis, the expression of the LXDD1 in human normal tissues and the size of the transcription of the LXDD1 representative gene were also determined using MTN (Multiple Tissues Northern Blots). The putative full-length cDNA of the EST-representative gene was cloned and analyzed by bioinformatics. In addition, differential reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of the novel gene in various cancer cell lines, primary lung carcinomas, and matched normal lung tissues. RESULTS: The full length cDNA with no homology to any reported genes in the database of GenBank was successfully cloned and named HLCDG1 (Human lung carcinoma deleted gene 1, GenBank accession number AF447582). A transmembrane protein with 166 amino acids was deduced to come from the open reading frame of the 3113 bp full-length cDNA, HLCDG1 gene was confirmed to be located at chromosome band 5q33 by alignment of electric polymerase chain reaction (e-PCR). CONCLUSION: HLCDG1 is a novel gene down-regulated in lung carcinoma, which may be involved in the development of lung carcinoma. PMID- 14558942 TI - [Differentially expressed genes in hepatocellular carcinoma of tree shrew induced by different factors]. AB - BACKGROUND & OBJECTIVE: Previous studies on differentially expressed genes in hepatocellular carcinoma (HCC) used to perform with para-cancerous tissues as normal control. However, the para-cancerous tissue of HCC is actually abnormal because they frequently contain hepatitis, cirrhosis, hyperplastic nodules or foci, etc. In order to explore the molecular mechanism and the responsible genes for hepatocarcinogenesis, through applying the HCC model of tree shrew (Tupaia belangeri chinensis), this study was designed to compare gene expression levels between HCC induced by different factors and their corresponding biopsies taken before HCC formation. METHODS: Tree shrews were divided into two groups. Group AFB(1) was fed with aflatoxin B1 (AFB(1)). Group AFB(1)+HBV was infected firstly with human hepatitis B virus (HBV) and then fed with AFB(1) as group AFB(1). Serum tests for HBV markers and liver biopsies were performed periodically during the experiment. After appearance of HCC, 2 HCC samples from each group and their corresponding 30th-week biopsies were tested respectively by cDNA microarray assay. The gene expression levels were compared between each HCC and the corresponding biopsies, and the differentially expressed genes from the two groups of HCC induced by different factors were analyzed. RESULTS: The incidence rates of HCC in group AFB(1) and group AFB(1)+HBV were 73.3% and 77.8%, respectively. A considerable number of genes in both groups showed changes in their expression levels, which were mainly up-regulated in group AFB(1) but down regulated in group AFB(1)+HBV. On the other hand, among the 588 checked genes (16 functional classifications) that were known related to human cancer, 11 genes were similarly expressed in all of the 4 HCC from the two animal groups. Most of these 11 genes belonged functionally to 3 types, namely "apoptosis-associated protein","DNA synthesis,repair and recombination proteins", and "growth factors, cytokines and chemokines". CONCLUSION: (1)HBV can affect AFB(1)-induced gene expression in certain extent. (2)The gene expression profiles of HCC induced by different factors are different. (3)The common differentially expressed genes in these two HCC groups are worthwhile for further study as the possibly responsible genes for hepatocarcinogenesis. PMID- 14558943 TI - [Role of eukaryotic initiation factor-4E (eIF-4E) in regulation of expression of NF-kappaB and its subsequent influence on transcription and activity of heparanase in human colon adenocarcinoma cell line]. AB - BACKGROUND & OBJECTIVE: Recent studies have revealed that some transcription factors may be translationally regulated by eukaryotic initiation factor-4E (eIF 4E) in human cancer. These modifications in the expression levels of the key transcription factors will alter the expression of some malignancy-related gene products at the transcriptional levels. The current study was designed to investigate the effect of eIF-4E on the expression and activity of NF-kappaB and observe how the activity level of NF-kappaB contributes, as a secondary effect, to the transcription of heparanase in human colon adenocarcinoma LS-174T cells. METHODS: In order to repress the expression of eIF-4E, a 20-mer antisense oligodeoxynucleotide (ASODN) targeted against the translation start site of eIF 4E mRNA was transfected into human colorectal cancer cell line LS-174T via liposome reagent, followed by assessment of the activity and the protein expression of NF-kappaB by an electrophoretic gel mobility shift assay (EMSA) and Western blot analysis respectively. The alterations of heparanase expression were examined by RT-PCR and Western blot analysis, and heparanase activity in LS-174T cells was measured by specific enzymatic activity test using radiolabeled heparan sulfate as the substrate and gel filtration chromatography for the analysis of the degradation product. RESULTS: The 20-mer ASODN against eIF-4E specifically and significantly inhibited eIF-4E expression at both transcriptional and translational levels, and the repression of eIF-4E gene expression was correlated with decreased expression levels and activity of NF-kappaB protein. Furthermore, this down regulation of the ubiquitous transcription factor NF-kappaB led to reduced transcription of the heparanase gene, and the transfected cells also showed a considerable decrease in heparanase protein and activity. CONCLUSION: These results suggest that eIF-4E play an important role in translational regulation of NF-kappaB expression in LS-174T cells.NF-kappaB is an essential factor in the regulatory mechanisms of heparanase gene transcription, and the suppressed NF-kappaB activity in LS-174T cells may significantly reduce the transcriptional expression of heparanase that consequently leads to decreased heparanase protein expression and enzymatic activity. PMID- 14558944 TI - [Study of anti-breast cancer activity of tumor infiltrating lymphocytes affected by dendritic cells]. AB - BACKGROUND & OBJECTIVE: Dendritic cell (DC) is the strongest antigen presenting cell(APC). It can present antigen to T lymphocytes in vivo and in vitro,and induce cytotoxic T lymphocyte(CTL) reactions.This study was designed to investigate the killing activity of tumor infiltrating lymphocytes (TILs) stimulated by dendritic cells on breast cancer cells in vitro. METHODS: DCs were isolated from peripheral blood of patients with breast cancer. DCs were stimulated by granulocyte/macrophage colony stimulating factor (GM-CSF), interleukin-4(IL-4), and tumor antigen. Then TILs were stimulated by DCs and their killing activity on autogenous breast cancer cells and Bcap-37 breast cancer cells in vitro were observed. RESULTS: TILs stimulated by DCs had very high killing activity on autogenous breast cancer cells and the killing rate was (85.76+/-2.93)%. The killing rate was higher obviously than that of TILs not stimulated by DCs and T lymphocytes stimulated by DCs or not on autogenous breast cancer cells, respectively [killing rates: (52.11+/-1.48)%, (51.35+/-1.46)%, and (3.59+/-0.25)%, respectively]. However, their killing activities on Bcap-37 breast cancer cells were lower [killing rates: (40.03+/-1.29)%, (22.09+/-0.87)%, (21.66+/-0.85)%, and (1.76+/-0.14)%, respectively]. CONCLUSION: The results indicate that DC from the patients with breast cancer can induce TIL to produce efficient and specific anti-breast cancer immune response. PMID- 14558945 TI - [Relationship between Helicobacter pylori infection and expression of c-myc, Bcl 2, and Bax protein in different gastric mucosa lesions]. AB - BACKGROUND & OBJECTIVE: Helicobacter pylori (HP) has been believed to be a carcinogen of gastric carcinoma. However, its mechanism was yet not clearly understood. This study was designed to investigate the relationship between HP infection and gastric epithelial cell proliferation as well as apoptosis in different gastric mucosa lesions and elucidate the probable mechanism of gastric carcinogenesis relating with HP infection. METHODS: A total of 272 cases were available for the study including 42 cases of chronic gastritis (CG), 46 cases of intestinal metaplasia I or II (IM I- II), 25 cases of intestinal metaplasia III (IM III), 21 cases of mild dysplasia (Dys I), 54 cases of modest or severe dysplasia (Dys II- III), and 84 cases of gastric cancer (GC). HP infection was detected by Warthin-Starry bacterium staining method and streptavidin-peroxidase (SP) immunohistochemical method. HID-AB(pH2.5)- PAS method was used to define the quality of mucus. The expression of c-myc, Bcl-2, and Bax were detected using SP immunohistochemical method. The chi-square test and the Fisher's exact probability test were used to compare the frequencies. RESULTS: (1)The expression of c-myc and Bcl-2 increased as gastric mucosa lesions developed from CG,IM,Dys to GC,but the expression of Bax decreased. The expression of c-myc was significantly higher in GC than that in Dys II- III and IM III(all P< 0.01), but the expression of Bax was significantly lower in GC than that in Dys II- III and IM III(P< 0.05 or P< 0.01). (2)The expression of c-myc in IM III and Dys II- III with HP infection was 62.50% and 66.67%,respectively, significantly higher than that without infection(11.11%,27.78%,all P< 0.05). The expression of Bax in CG, IM I- II and IM III with HP infection were 87.10%, 81.25%, and 62.50%, respectively, significantly higher than those without infection (54.55%, 42.86%, 11.11%, all P< 0.05). Furthermore HP infection was associated with the expression of Bcl-2 in IM III, Dys II- III and GC (P< 0.05 or P< 0.01). CONCLUSION: HP infection can cause serious imbalance between cell proliferation and apoptosis in the precancerous lesions (IM III and Dys II- III), giving chances for gastric carcinogenesis. PMID- 14558946 TI - [Role of tissue factor pathway inhibitor-2 in ovarian tumor migration and invasion]. AB - BACKGROUND & OBJECTIVE: Human tissue factor pathway inhibitor-2 (TFPI-2) is a newly discovered serine protease inhibitor, which inhibits plasmin, trypsin, matrix metalloproteinase (MMPs), but not urokinase, tissue-type plasminogen activators and thrombin. Earlier studies have shown that the production of TFPI-2 is downregulated during the progression of various cancers. The aim of this study was to elucidate the relationship between TFPI-2 expression and ovarian tumor migration and invasion. METHODS: Human TFPI-2 expression vector pBos-Cite neo/TFPI-2 was transfected into ovarian tumor cells line A2780. After the transfected cells were screened by G418, transfected and nontransfected cells were examined for TFPI-2 mRNA and protein by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. The number of transfected or nontransfected cells passing through membrane of Boyden chamber was counted as the basis assessing tumor cells migratory and invasive behaviors. RESULTS: Expression of mRNA and protein of TFPI-2 were confirmed in transfected cells. In invasion assay, the number of TFPI-2-expressing cells to traverse a Matrigel-coated membrane was obviously decreased compared with that of nonexpressing cells (59.3+/-6.5 versus 109.7+/-5.5, P< 0.01); while in migration assay, no significant difference was observed between transfected and nontransfected cells (114.7+/-8.6 versus 127.3+/-7.1, P >0.05). CONCLUSION: Expression of TFPI-2 may strongly inhibit the invasive ability of ovarian tumor cells in vitro, but has no effect on the migratory ability, which provides an experimental basis for treating human ovarian tumor with gene therapy. PMID- 14558947 TI - [Effect of selective cyclooxygenase-2 inhibitor celebrex on expression of vascular endothelial growth factor (VEGF) in pancreatic carcinoma]. AB - BACKGROUND & OBJECTIVE: The previous study has identified that cyclooxygenase-2 (COX-2) may have a close relation with tumor genesis, particularly with digestive tract tumors, and its inhibitor can exert the chemoprevention role on carcinogenesis. This study was designed to investigate the effect of celebrex, a selective cyclooxygenase-2 inhibitor, on the expression of vascular endothelial growth factor (VEGF) in pancreatic carcinoma of xenografted nude mice induced by pancreatic carcinoma PC-3 cell lines. METHODS: The effect of celebrex on tumor growth was observed.The expression of VEGF in the tumors was determined by reverse transcription polymerase chain reaction (RT-PCR), Western blot analysis, and enzyme-linked immunosorbent assay (ELISA). RESULTS: Average tumor volume and tumor weight from control mice were 0.438+/-0.052 cm(3) and 0.552+/-0.064 g as compared with 0.215+/-0.038 cm(3) and 0.244+/-0.042 g from treated mice (inhibition rate:51.6%,P< 0.05). VEGF expression was significantly down-regulated in the celebrex-treated tumors. ELISA revealed that the expression levels of VEGF were 1.11+/-0.11(microg/g) in control mice and the 0.66+/-0.11(microg/g) in the treated mice. The inhibition rate of VEGF was 40.6% (P< 0.05). CONCLUSION: COX-2 may play an important role in the angiogenesis of pancreatic carcinoma. The selective COX-2 inhibitor, celebrex, can result in the inhibition of angiogenesis and tumor growth. PMID- 14558948 TI - [Different sensitivity of two chronic myeloid leukemia cell lines to interferon alpha]. AB - BACKGROUND & OBJECTIVE: The clinical observation has shown that interferon alpha(IFN-alpha) is one of the most effective therapeutic agents for the malignancies of hemopoietic system and lymphoma. However, IFN-alpha can only induce about 70-80% of the patients with chronic myeloid leukemia (CML) to get hematological remission. The mechanism by which various CML cases respond differently to IFN-alpha is still unclear. METHODS: (1)The effects of IFN-alpha in different concentrations (100, 500, 1,000, 5,000, and 10,000 U/ml) on growth of the two CML cell lines were detected by MTT assay,semisolid colony formation and trypan-blue staining for the cells in liquid culture. (2)The cell apoptosis was examined by flow cytometry(FCM),fluorescence microscopy and gel electrophoresis analysis for DNA fragmentation in 48 hours after IFN-alpha (1,000 U/ml) induction of both KT-1/A3 and K562 cells. (3)The expression levels of bcr/abl chimeric genes were analyzed by relative quantitative RT-PCR at 48 hours after cultivation of both KT-1/A3 and K562 cells with IFN-alpha in 1,000 U/ml. RESULTS: (1)The growth inhibition of KT-1/A3 cells was dose-dependent in IFN alpha from the concentration of 100 U/ml to 10,000 U/ml. (2)Having been induced with IFN-alpha in 1000 U/ml for 48 hours, the apoptosis rate of KT-1/A3 cells raised from 3.29% to 11.8% and the expression level of bcr/abl chimeric gene in this cell line declined to 66.7% as compared with those of the control. (3)The growth and apoptosis rate as well as bcr/abl gene expression level of K562 cells were not significantly affected by IFN-alpha. CONCLUSION: Different populations of CML cells shows different sensitivity to IFN-alpha. PMID- 14558949 TI - [Roles of VEGF-C and its receptor Flt-4 in proliferation and metastasis of primary breast cancer]. AB - BACKGROUND & OBJECTIVE: Vascular endothelial growth factor-C(VEGF-C) is a member of VEGF family and the only factor that can combine receptor VEGFR-3 (fms-like tyrosine kinase, Flt-4) located at endothelium of lymphatic vessel and modulates the physiological function of lymphatic vessel. Previous study showed that VEGF C/Flt-4 system play a modulating role in metastasis of many kinds of tumors, but there were few reports about its function for primary breast cancer at home and abroad. The objective of this study was to identify the function of VEGF-C/Flt-4 system in proliferation and metastasis of primary breast cancer and its significance. METHODS: A series of 101 primary breast cancer specimens were detected for the expression of VEGF-C, Flt-4,and PCNA by Immunohistochemical methods. RESULTS: Among 101 cases of breast cancer, the positive rate of VEGF-C was 93.1% (94/101), the positive rate of flt-4 was 86.1% (87/101). With the increase of the expression of VEGF-C, the positive index of flt-4 increased (r=0.816,P< 0.001). The positive rate of PCNA was 88.8% (89/101). With the increase of the expression of VEGF-C, the proliferation activity of PCNA was stronger (r=0.673,P< 0.001). The positive index of VEGF-C in lymph node metastases group (61.89+/-17.79) was significantly higher than that of no lymph node metastasis group (44.28+/-17.87)(P< 0.05). With the increase of VEGF-C protein level, the number of flt-4 positive vessels increased and significant differences among these groups were observed (P< 0.001). The number of flt-4 positive vessels in lymph node metastases group (15.55+/-3.63)was significantly higher than that of no lymph node metastasis group (10.71+/-2.90 ) (P< 0.05). CONCLUSION: VEGF-C and Flt-4 are overexpressed in primary breast cancer and related to lymph node metastasis. VEGF-C can promote proliferation of breast cancer cell. VEGF-C/flt-4 system can promote vasculogenesis in stroma of breast cancer. The number of Flt-4 positive vessels is closely related to lymph node metastasis. PMID- 14558950 TI - [Expression of bcl-2, c-erbB2, and p53 protein in cervical epithelial carcinogenesis]. AB - BACKGROUND & OBJECTIVE: Less research of molecular biology have been reported on cervical carcinogenesis. So the authors designed this study to evaluate the expression and clinical significance of antiapoptosis gene bcl-2, oncogene c erbB2, and tumor suppressor gene p53 in the progression of this kind of cancer. METHODS: Using S-P immunohistochemical technique, the authors examined the expression of bcl-2, c-erbB2, and p53 in 48 squamous cell carcinoma of the cervix, 42 cervical intraepithelial neoplasia (CIN), and 20 normal cervical tissues. RESULTS: The expression rates of bcl-2, c-erbB2, and p53 were 72.9%(35/48), 60.4% (29/48), and 58.3% (28/48) in carcinomas, 61.9% (26/42), 38.1% (16/42), and 26.2% (11/42) in CIN, 20.0% (4/20), 0.0% (0/20), and 0.0% (0/20) in normal cervices, respectively. The overexpression of bcl-2 protein in carcinomas and in CIN was remarkably higher than that in normal cervices (P< 0.01), and the difference between carcinomas and CIN was not significant (P>0.05). There were significant differences for c-erbB2 and p53 between carcinomas and CIN and normal cervices, respectively (P< 0.05). There were significant differences for bcl-2, c-erbB2, and p53 between CIN3 and CIN1-2, respectively (P< 0.05). Expression of bcl-2 and p53 were correlated with histological type, tumor grades, and clinical stages (P< 0.05). Expression of bcl 2 was lower keratinized than nonkeratinized and was lower as tumor grades and clinical stages increased (P< 0.05). It was not associated with lymph node metastasis (P >0.05). However, the expression of p53 was contrary with bcl-2. Expression of c-erbB2 was not associated with histological type, tumor grades, clinical stages, and lymph node metastasis (P >0.05). CONCLUSION: In cervical carcinogenesis, the overexpression of bcl-2 protein may be activated in the early stage of pathogenesis of cervical carcinoma. Expression of c-erbB2 was the signal of cell malignant change. Expression of p53 was associated with the pathogenesis and development of cervical carcinoma. PMID- 14558951 TI - [Expression of transporter associated with antigen processing (TAP) in childhood acute leukemia and its clinical significance]. AB - BACKGROUND & OBJECTIVE: Transporter associated with antigen processing(TAP) participates in immune surveillance, so it is probably relevant to carcinogenesis. Investigation of expression of TAP and its clinical significance in childhood acute leukemia will be helpful to clarify pathogenesis and to develop immunotherapy strategy. METHODS: RT-PCR analysis was used to detect the expression of TAP1 and TAP2 in leukemia cells from bone marrow in 34 inpatients with primary acute lymphoblastic leukemia (ALL), 15 inpatients with relapsed ALL, 20 inpatients with acute medullary leukemia (AML), and 20 surgical inpatients without systematic disorders as control. And then,the absorbance (A) values of expanded bands were measured by digital imaging analyzer and relative A values were worked out based on A value of GAPDH (positive inside control). RESULTS: The relative A values of TAP1(0.448+/-0.167 and 0.169+/-0.021,respectively) and TAP2(0.196+/-0.180 and 0.112+/-0.020, respectively) in primary ALL group and relapsed ALL group were lower than those in control group (P< 0.01). The A values of LMP2, LMP7, and PA28alpha (991.4+/-532.7, 686.3+/-663.8, and 2022.3+/-1622.3, respectively) in relapsed ALL group were lower than those in control group (P less than 0.01,0.01 and 0.05, respectively). The A value of LMP2 in relapsed group was lower than that in control (P< 0.01). The A values of LMP7 for the cases with no remission and relapse were both lower than that for the cases with constant complete remission in primary ALL group (P< 0.01). The relative A value of TAP2 in AML group was lower than that in control group (P< 0.05). The relative A value of TAP1 in relapsed ALL group was lower than that in primary ALL group (P< 0.05). In primary ALL group, the relative A value of TAP1 (0.215+/-0.159) for cases with relapse (6/34 cases) was lower than that (0.462+/-0.189) for those with constant complete remission (24/34 cases) (P< 0.05). CONCLUSION: There exists decreased expression of TAP in both childhood ALL and AML, which probably contributes to the escape of leukemia cells from immune surveillance. Decreased expression of TAP1 subunit is probably related to the relapse of ALL. PMID- 14558952 TI - [Radical cystectomy with sparing partial prostate for invasive bladder cancer]. AB - BACKGROUND & OBJECTIVE: Classical radical cystectomy for invasive bladder cancer removes the urinary bladder and prostate completely and was associated with high incidence of post-operative impotence and urinary incontinence. It is uncertain whether prostate excision has any effects on the prognosis of patients undergone cystectomy if the prostate is not involved by the tumor. Sparing the prostate in radical cystectomy might improve potency and urinary control. This paper was to report our experience of sparing partial prostate in radical cystectomy for ten male patients. METHOD: Transurethral resection of the prostate and radical cystectomy with sparing partial prostatic capsula were carried out in 10 male patients with invasive bladder transitional cell carcinoma. Intestinal neobladder, anastomosed with the residual capsula, were used for reconstruction of the lower urinary tract. Clinical outcome of these patients including tumor control,urinary continence, and sexual status was evaluated. RESULTS: Mean follow up was 9 months (range 3-12 months). Postoperative pathological stage was T2N0M0 in all patients. Nine patients survived free of disease. One patient who had recurrence of poorly differentiated transitional cell carcinoma developed metastasis in bone and lymph nodes two months after cystectomy. All patients voided smoothly. Nine patients were continent day and night. One patient had mild incontinence. Of 8 patients with previously sexual function, 6 maintained potency. CONCLUSION: Satisfactory urinary control and sexual potency could be achieved in radical cystectomy with sparing partial prostate. Further study is needed about the long-term effects of sparing prostate in cystectomy on tumor control. PMID- 14558953 TI - [Retrospective analysis of effectiveness of tonsillar squamous cell carcinoma: report of 108 cases]. AB - BACKGROUND & OBJECTIVE: There is a controversy in selecting either radiotherapy plus surgery reserved as salvage (Salvage S) or the planned surgery plus preoperative radiotherapy (Planned R+S) as the treatment modality for the tonsillar squamous cell carcinoma. Among stage III- IV patients, the tumor size and the lymph node metastasis were insufficient as the prognostic predictors. The authors retrospectively reviewed the patients with this disease who were treated in Cancer Hospital of Chinese Academy of Medical Sciences to analyze the indications and prognosis of the two treatments. METHODS: A total of 108 patients with biopsy-proven tonsillar squamous cell carcinoma were reviewed from 1984 to 2000. The patients were divided into Salvage S group (group A, 83 cases) and Planned R+S group (group B, 25 cases). Treatment outcome and the surgery related oral dysfunctions were compared in the two groups. The relationship between the response to radiotherapy and the survival rate was analyzed. RESULTS: The composite resection that led to oral dysfunction was undertaken for 20 (24.1%) of group A patients and for 22 (88.0%) of group B patients. The 5-year survival rates were 59.3% and 55.4% in group A and group B patients, respectively (P=0.706). The 5-year survival rates were 73.0% and 76.9% in the patients whose lymph nodes and surgical specimen showed obvious response to radiotherapy,respectively. CONCLUSION: The two treatment modalities reached similar survivals. However, compared with the planned surgery, 60% of composite resection would be withdrawn if surgery reserved as salvage, being benefit to the oral function. The responses to radiotherapy in the lymph nodes and the surgical specimen were good predictors of prognosis. PMID- 14558954 TI - [Effects of ulinastatin on interleukin-8 during one-lung ventilation in surgery]. AB - BACKGROUND & OBJECTIVE: Ulinastatin is a kind of broad- spectrum hydrolase inhibitors purified from urine of healthy males.There was no data showing whether ulinastatin could affect alveolar inflammatory reactions.This study was designed to investigate the changes of pulmonary alveolar interleukin-8 during one lung ventilation (OLV) in surgery and the effects of ulinastatin on alveolar IL-8. METHODS: Sixty patients,ASA II- III,who underwent lobectomy, were randomly divided into two groups [30 patients in each group,surgery group(II) and control group (I)].Patients in surgery group were treated with ulinastatin 5,000 u/kg preoperatively. Alveolar IL-8 was harvested by bronchoalveolar lavage at 0, 1, and 2 hours after one lung ventilation and at the end of surgery. The changes of concentration of IL-8 in bronchoalveolar lavage fluid were analyzed statistically. RESULTS: (1)Alveolar interleukin 8 was significantly higher at 2 hours after OLV [(160.85+/-46.58)ng/L] than before OLV [(128.26+/-35.35)ng/L] in control group. The results indicated that OLV and/or the stimulation of surgery could induce the pulmonary inflammatory reaction. (2)The concentrations of alveolar IL-8 at 2 hours [(140.32+/-41.25)ng/L] after OLV and at the end of surgery [ (156.74+/-43.58) ng/L] were significantly lower in surgery group than those in control group [(160.85+/-46.58)ng/L and (172.41+/-52.60)ng/L] respectively(P< 0.05). CONCLUSION: Ulinastatin could alleviate the pulmonary alveolar inflammatory reactions in OLV. PMID- 14558955 TI - [Expression of nm23 and proliferating cell nuclear antigen (PCNA) in human brain gliomas and their significance]. AB - BACKGROUND & OBJECTIVE: Brain gliomas seldom undergo extracranial metastasis. Local recurrence is the main reason of tumor patient's death. Therefore, it is important to detect tumor biological features through determination of gene expression. This study was designed to investigate the expression of nm23 (non metastasis gene, nm23)and PCNA (proliferating cell nuclear antigen) and evaluate the malignancy, recurrence, and prognosis of the tumor. METHODS: In 50 specimens of different malignant gliomas,the expression of nm23 and PCNA were examined using SP immunohistochemical staining. RESULTS: (1)The label indexes of nm23 and PCNA in low-grade gliomas were 3.40+/-0.27 and 3.60+/-0.05, respectively; while the label indexes of nm23 and PCNA in high-grade gliomas were 1.72+/-0.18 and 6.20+/-0.23, respectively.There was significant difference between the two groups(P< 0.05). (2)The positive rates of nm23 and PCNA were 56% (14 cases) and 64% (16 cases) in 25 cases of low-grade gliomas, while the positive rates of nm23 and PCNA were 12% (3 cases) and 88% (22 cases) in 25 cases of high-grade gliomas. There was significant difference between the two groups (P< 0.05). (3)The positive rates of nm23 and PCNA were 0% (0 cases) and 100% (9 cases) in 9 cases of recurrent gliomas, while the positive rates of nm23 and PCNA were 50%(34 cases) and 50%(4 cases) in 8 cases of non-recurrent gliomas. There was significant difference between the two groups (P< 0.05). (4)The label indexes of nm23 and PCNA in gliomas were inversely correlated (r=-0.5335,P< 0.001). CONCLUSION: (1)The expression of nm23 is inversely correlated with the malignancy of gliomas,i.e.the lower expression indicates the higher malignancy. (2)The expression of PCNA is associated with the increased malignancy. (3)Both nm23 and PCNA may be useful biological markers to evaluate the malignancy and prognosis of patients with gliomas. PMID- 14558956 TI - [Surveillance of cerebral blood circulation changes by transcranial Doppler ultrasonography in patients with nasopharyngeal carcinoma after radiotherapy]. AB - BACKGROUND & OBJECTIVE: Radiotherapy is the first choice for patients with nasopharyngeal carcinoma (NPC), but radiation encephalopathy affects the patients' quality of life significantly. Some research results have showed that the brain damage was associated with radiation-induced cerebral vascular injury. The study was designed to determine the cerebral blood circulation changes in NPC patients pre- and post- radiotherapy using transcranial Doppler ultrasonography (TCD), and to evaluate its value in the early diagnosis and treatment of radiation encephalopathy. METHODS: The blood flow velocity of intra-carotid artery (ICA), middle cerebral artery (MCA), and basal artery (BA) were measured by TCD in 38 patients at the beginning, the end of radiotherapy, 3 months and 3-5 years after radiotherapy respectively. The flow velocity results of each cerebral artery in each examination time after radiotherapy were compared with the findings before treatment correspondingly. RESULTS: The velocity of both side ICA kept stable during all the follow-up time. The mean value of velocity of the left and right MCA raised obviously, from 53-54 cm/s before treatment to 59-60 cm/s at the end of radiotherapy (P< 0.05). It resumed to 54 cm/s after 3 months, and went up again to 57-58 cm/s during 3-5 years after radiotherapy, but there was no remarkable difference in statistics (P >0.05). The mean velocity of BA was 30 cm/s before irradiation, and raised to 35 cm/s at the end of treatment (P< 0.05). After 3 months, it descended to 32 cm/s (P >0.05), and attained to 34 cm/s at 3-5 years after radiotherapy (P< 0.05). CONCLUSION: During several stages of radiotherapy, the flow velocity of MCA and BA were raised obviously in NPC patients. PMID- 14558957 TI - [Clinical study in prophylactic use of chinese medicine to prevent chemoradiotherapy induced mucositis in nasopharyngeal carcinoma]. AB - BACKGROUND & OBJECTIVE: Oropharyngeal mucositis is the most common acute non hematology toxicity in nasopharyngeal carcinoma (NPC) treated with radiotherapy, especially in the concomitant chemoradiotherapy of local advanced NPC patients. This study was designed to observe the effect of traditional Chinese medicine against acute oropharyngeal mucositis from chemoradiotherapy in patients with local advanced NPC. METHODS: A total of 101 patients in stage III- IVa (Fuzhou 1992) were enrolled into this prospective randomized clinical trial. The cases were divided into treatment group (52 cases) and control group (49 cases). The median doses were 70.31+/-1.21 Gy for the treatment group and 70.78+/-1.95 Gy for the control group, respectively. Chemotherapy was concomitant with radiotherapy [single agent cisplatin (DDP,30 mg/m(2)) 6 times from first to sixth week of radiotherapy duration]. The patients of treatment group took 5-8 times of Chinese medicine daily and those of control group took 5-8 times of Dobell's solution daily.The observation indices included the degree of oropharyngeal and hematological toxicity, radiotherapy duration, and curative effect. RESULTS: (1)Oropharyngeal toxicity: there was no 0 degree oropharyngeal toxicity in both groups, I degree toxicity in 29 cases (55.77%) and 2 cases (4.08%), II degree toxicity in 18 cases (34.62%) and 17 cases (30.69%), III degree toxicity in 5 cases (9.62%) and 22 cases (44.89%), IV degree toxicity in 0 case (0%) and 8 cases (16.33%); there was statistical significance of difference between the two groups (P=0.000). (2)Hematological toxicity: there was no IV degree hematological toxicity in both groups. WBC toxicity (Z=-0.604,P=0.546), neutrophilic granulocyte toxicity (Z=-0.226,P=0.821), and platelet toxicity (Z=-0.099,P=0.922) were compared; there was no statistic significance of difference (P >0.05); (3)Radiotherapy duration: The radiotherapy duration of two groups were 49.79+/ 1.83 days and 51.00+/-4.46 days; there was no statistic significance of difference (P=0.082). (4)Curative effects: The curative effects of two groups were:complete remission (CR) 76.92% (40/52) and 69.39% (34/49), partial remission (PR) 23.08% (12/52) and 30.61% (15/49); there was no statistical significance of difference (P=0.392). CONCLUSION: Chinese medicine was effective in reducing acute oropharyngeal toxicity resulting from chemoradiotherapy in patients with local advanced NPC. Furthermore, the apparent side effects of Chinese medicine in this study have not been observed. Chinese medicine treatment did not affect the short-term clinical outcome. PMID- 14558958 TI - [Effectiveness evaluation and prognostic factor analysis in patients with minor salivary gland carcinoma of the hard palate]. AB - BACKGROUND & OBJECTIVE: Minor salivary gland carcinoma of the hard palate is rare, and its long-term survival rate is high, making it difficult to evaluate the prognostic factors and the efficacy of treatment. This study was designed to evaluate the efficacy of treatment and investigate the prognostic factors of minor salivary gland carcinoma of the hard palate. METHODS: Ninety-six cases of minor salivary gland carcinoma of the hard palate hospitalized in Cancer Center, Sun Yat-sen University, from 1964 to 2001 were reviewed retrospectively. The cumulative survival rate was analyzed by Kaplan-Meier method. The factors were compared using the log-rank test. The influencing factors were screened by Cox proportional hazards model. RESULTS: The 5- and 10-year overall survival rates were 65.60% and 47.90%, respectively. The 5- and 10-year disease-specific survival rates for the patients treated with surgery (53 cases) and surgery + radiotherapy (35 cases) were 73.14%, 67.40% and 66.58%,46.60%,respectively. Multivariate analysis showed that age >or=50 years old,tumor size >or=3 cm,surgical margin status,and recurrence were independently associated with decreased survival rates (P< 0.05). CONCLUSION: Surgery or surgery dominated multi-modality therapy was the principal treatment modality for minor salivary gland carcinoma of the hard palate. Age >or=50 years old, tumor size >or=3 cm, surgical margin status, and recurrence are independent factors affecting the prognosis. PMID- 14558959 TI - [Expression of P27 protein and cyclin E in colon cancer]. AB - BACKGROUND & OBJECTIVE: P27 protein and cyclin E were negative cell cycle regulators. Until the present, the influence of P27 protein and cyclin E on progression of colon cancer was unclear. The aim of this study was to observe the expression features of P27 protein and cyclin E in the tissues of colon neoplasms, and to investigate the relationship between colon neoplasms and tumor special growth factor (TSGF). METHODS: Sixty-nine cases of samples included 23 normal tissues, 28 colon polyps (13 inflammatory polyps and 15 adenomatous polyps), and 18 colon carcinomas. The location and expression of P27 protein and cyclin E were determined using immunohistochemical method in all samples. These samples were diagnosed using formal pathological techniques simultaneously; the relationship between colon neoplasms and TSGF was also investigated. RESULTS: The positive signal of P27 and cyclin E was found mainly in the cytoplasm and extracellular matrix of normal colon tissues, inflammatory polyps, and adenomatous polyps. Less amount of positive expression product of P27 protein and cyclin E was observed in colon carcinoma cells; and the positive signal was only located in the cytoplasm of gland-like cells. The content of TSGF in colon carcinoma tissues was significantly higher than that in normal tissues (117.3+/ 57.02 versus 64.16+/-27.5,P< 0.01), but there was no significant difference between colon carcinoma tissues and inflammatory polyp tissues (117.3+/-57.02 versus 92.5+/-47.9,P >0.05). CONCLUSION: P27 protein and cyclin E participate in the adjustment process of colon neoplasm occurrence and progression. The reduced expression of P27 protein and cyclin E may indicate the possibility of colon carcinoma. PMID- 14558960 TI - [Value of acetic acid smear test for cervical cancer screening]. AB - BACKGROUND & OBJECTIVE: At present, the incidence of cervical cancer is still high in less developed areas of China; one of the reasons is that cytology in screening for cervical cancer is not feasible in these areas. The aim of this study was to analyze the sensitivity and specificity of acetic acid smear test as a cervical cancer screening test, and evaluate the value of this method for cervical cancer screening. METHODS: Acetic acid smear test was performed on 1997 previously unscreened women aged 35-45 years in Xiangyuan County, Shanxi Province. All women had cervical cytology, testing for high-risk types of human papillomavirus DNA, colposcopy, an endocervical curettage and at least five cervical biopsies. Biopsies were done at the lesions, or one biopsy at 2, 4, 8, 10 o'clock at the squamocolumnar junction in each normal quadrant. RESULTS: According to histological diagnosis, 43 women had CIN II, 31 had CIN III, and 12 had invasive carcinoma. In two women only the endocervix was positive. Smear test yielded normal results in 1445 women (72.4%), low-grade squamous intraepithelial lesions (LSILs, CIN I) in 525 (26.3%), high-grade squamous intraepithelial lesions (HSILs, CIN II- III) in 21 (1.1%), and cancer in 6 (0.3%). With abnormal visual inspection defined as LSILs or worse, the sensitivity of acetic acid smear test for HSILs or worse was 70.9% (61 of 86), the specificity was 74.3% (1420 of 1911). The sensitivity was 64.9% for smaller lesions (37 of 57), and 88.9% for larger lesions (24 of 27) (P=0.03). The sensitivity of colposcopy for high-grade SILs or worse was 81.4% (70 of 86), the specificity was 76.5% (1462 of 1911). CONCLUSION: This study showed the high specificity and sensitivity of acetic acid smear test as a primary screening test for cervical cancer. Acetic acid smear test and colposcopy have similar specificity profiles for CIN II or more severe lesions. Acetic acid smear test as a primary screening test will be possible and effective with the benefits of low-cost and simple test in low-resource settings. PMID- 14558961 TI - [Combined surgical treatment for limited small cell lung cancer: clinical analysis of 51 cases]. AB - BACKGROUND & OBJECTIVE: Small cell lung cancer (SCLC) is usually widely disseminated at the time of diagnosis due to its rapid growth. Therefore chemotherapy is the predominant form of treatment for SCLC. However, chemotherapy normally ended in failure as a consequence of recurrence of primary tumor or mediastinal lymphatic nodes. The objective of this study was to investigate whether combined surgical treatment for SCLC can control primary tumor more effectively and increase the survival of the patients. METHODS: Fifty-one cases of limited SCLC treated with surgery from May 1981 to May 2001 in Cancer Center, Sun Yat-sen University were reviewed retrospectively. RESULTS: According to 1997 international staging system, there were 7 patients with stage IB(13.7%), 5 with stage IIA(9.8%), 15 with stage IIB (29.4%), 20 with stage IIIA (39.2%) and 4 with stage IIIB (7.8%). Twelve patients received pneumonectomy, 35 lobectomy, 3 segmentectomy, and 1 exploration. Thirteen patients were treated with surgery alone and 38 of them combined with 2-6 cycles of pre- or post- operative chemotherapy. The median survival time was 20.8 months. The overall 1-, 3-, and 5 year survival rates were 43.0%, 25.9%, and 20.1%, respectively. The 1-, 3-, and 5 year survival rates for patients with stage IB were 100.0%, 80.0%, and 30.0% respectively, with stage II were 79.7%, 39.8%, and 34.5%, respectively, with stage III were 52.4%, 21.8%, and 5.46%, respectively. The 1-,3-, and 5-year survival rates for patients treated by surgery combined with chemotherapy were 77.5%, 38.5%, and 23.8% and that of surgery alone were 41.7%, 16.7%, and 8.3%, respectively (P< 0.01). CONCLUSION: Combined surgical treatment can increase the survival rate of SCLC more effectively and should be considered as an important modality in multi-disciplinary treatment of limited small cell lung cancer. PMID- 14558962 TI - [Clinical analysis of 44 cases of recurrent thyroid carcinoma]. AB - BACKGROUND & OBJECTIVE: Although the most thyroid cancers are well-differentiated low malignant neoplasms and their prognoses are relatively good generally if they were treated routinely, there are still a few cases of this tumor recurred. This study was designed to sum up and analyze the cause, treatment, and prognosis of these cases to seek for the cause and the proper treatment method of the recurrent cases. METHODS: From 1980 January to 1996 January,44 cases of the recurrent thyroid carcinoma of the total 764 cases of thyroid carcinoma were treated in Department of Head & Neck,Cancer Institute & Hospital, Affiliated Tianjin Medical University. The clinical and follow-up data of these 44 cases were analyzed retrospectively. RESULTS: Forty-four cases of the recurrent thyroid carcinoma accounted for 5.76%(44/764) of 764 cases of thyroid carcinoma treated. Thirty-eight cases of these 44 cases were differentiated thyroid carcinoma (28 cases were papillary thyroid carcinoma,7 cases were follicular thyroid carcinoma, 3 cases were medullary thyroid carcinoma and 6 cases were anaplastic thyroid carcinoma).The percentages of them accounted for the same histopathologic pattern thyroid carcinoma treated in the hospital at the same time were 4.1% (28/676), 18.9%(7/37), 8.1% (3/37), and, 42.9% (6/14), separately. Thirty-three of these 44 cases were lymphatic recurrence (75.0%). In spite of 5 cases were given radiotherapy only, the other 39 cases were treated surgically with extensive dissection of recurrent lesions and post-operative radiotherapy. After treatment, the 3-year survival rates were 85.7%, 71.4%, 100.0%, and 0; the 5-year survival rates were 60.7%, 57.0%, 66.7%, and 0, and the 10-year survival rates were 35.7%, 42.9%, 33.3%, and 0, respectively. CONCLUSION: The clinical and subclinical metastatic lymph nodes left in the previous operations were the main cause of the recurrence of thyroid carcinoma. The recurrent thyroid carcinoma should be treated with surgical operation mainly, as most of them may have a relatively good prognosis after the treatment. PMID- 14558963 TI - [Advances on zeta protein encoded by Epstein-Barr virus (EBV) BZLF1 gene]. AB - Zeta protein was encoded by BZLF1 gene which is one of Epstein-Barr virus (EBV) immediate early (IE) genes. The protein is a member of activator protein 1 (AP-1) super family that can bind to cellular AP-1 response element. Zeta protein is associated with lots of human diseases, including nasopharyngeal carcinoma (NPC). So detection of Zeta protein or its antibody can be used in disease diagnosis and prognosis assessment. Zeta protein can repress human immune system, interfere with cellular signal transduction, affect cell cycle progress, and induce cell apoptosis. At the same time, its expression can be affected by some cellular components. Here we review the new advances on BZLF1 gene structure, expression and relationship to cells, together with association with EBV associated diseases. PMID- 14558964 TI - [Autofluorescence bronchoscopy for early localization of lung cancer]. AB - Autofluorescence bronchoscopy (AFB) is more sensitive than conventional white light bronchoscopy in localizing small premalignant and early malignant lesions involving the large airways, particularly when applied to high- risk patients. The applications of AFB include localization of roentgenographically occult lung cancer, detection of synchronous tumors, estimation of the extent of field cancerization,chemoprevention studies, etc. This review deals with the history of AFB and recent clinical studies in AFB and discusses its current value as a research tool. PMID- 14558965 TI - [Biocompatibility of poly-l-lysine-modified silica nanoparticles]. AB - BACKGROUND & OBJECTIVE: Poly-l-lysine-modified silica nanoparticle(PMS-NP) was a novel non-viral vector for gene delivery. The current study was designed to evaluate the biocompatibility of PMS-NP for its further utilization in vivo. METHODS: Cell transfection and flow cytometry were used to elucidate the delivery efficiency of plasmid DNA and antisense ODN mediated by PMS-NP in the presence of serum-containing medium. Subsequently, the biocompatibility of PMS-NP in vivo was evaluated using filtration assay of plasma proteins and erythrocyte aggregation assay. RESULTS: The abilities of PMS-NP to deliver plasmid DNA and antisense ODN in vitro clearly decreased in the presence of serum-containing medium. PMS NP/DNA(ODN)complexes bound plasma proteins and triggered erythrocyte aggregation. CONCLUSION: PMS-NP might interact with plasma proteins, resulting in decreased transfection efficiency in vitro. And filtration assay of plasma proteins and the erythrocyte aggregation assay demonstrated that the interaction of PMS-NP with plasma proteins and erythrocytes might play a negative role in gene transfection efficiency in vivo. And its biocompatibility needs to be further improved. PMID- 14558966 TI - [Analysis of clinical application for various types of esophageal stents made in China]. AB - BACKGROUND & OBJECTIVE: Dysphagia is the main clinical symptom of the patients with esophageal carcinoma. The stent placement in esophagus is an effective method to resolve dysphagia.There are many types of stents. Through comparing observation of the stents made in China, which is the stainless steel "Z" stent, the Ni-Ti wire mesh-like stent, and the covered or uncovered-stent, the authors analyzed the clinical application of various types of esophageal stents in the treatment of malignant esophageal stricture. METHODS: The various types of esophageal stents were implanted into 40 patients with esophageal carcinoma. 13 cases for stainless steel "Z" stents, 27 cases for Ni-Ti wire mesh-like stents(24 cases were the covered stents and 16 cases were uncovered). All the patients were not operated with the dilation of balloon before stent placement. RESULTS: The esophageal stents were successfully implanted into 40 patients without complication of operation. Stainless steel "Z" stent were all dilated immediately at the time of operation, and all the 13 patients had obvious intra-breastbone pain after operation, which lasted 10-20 days. Although Ni-Ti wire mesh-like stents could not dilate completely in operation, all these can be dilated completely after 1 to 3 days; nine cases of them had obvious intra-breastbone pain, which lasted 3-5 days. In all these, 7 cases were the covered-stents and 2 cases uncovered. In 9 cases, tracheoesophageal fistulas were blocked completely. There was barium sulphate retention between the covered-stent implanted to the patients without tracheoesophageal fistula and the dilated esophagus. CONCLUSION: The clinical adaptability of the Ni-Ti wire mesh-like stent is better than that of the stainless steel "Z" stent. For the patients with tracheoesophageal fistula, the first choice is covered-stent, while for the patients without tracheoesophageal fistula, uncovered-stent is better. The dilation of balloon is not necessary before the stent placement. PMID- 14558967 TI - [The importance of transplants]. PMID- 14558968 TI - [Local analgesia in postoperative cleansing of head and neck infected wounds]. AB - INTRODUCTION: After debridement of head and neck abscess, patients require multiple post-surgical cleansing procedures that produce mild or severe pain because are performed without any effective analgesia. Sedation techniques could not be applied at every cleansing process and even local anesthesia injected into the tissues during the procedure is contraindicated. MATERIAL AND METHODS: We present the results of pain control obtained in 600 cleansing procedures performed by irrigating an anesthetic solution over wound tissues exposed after surgical head and neck abscess debridement. RESULTS: All patients had previous surgical debridement of head and neck abscess. All were irrigated with lidocaine solution because dressings were eliminated during every cleansing process. During 5 days of follow-up, at the first and second day only 12.5% of cases reported severe pain when dressing materials were eliminated; at the third day, only one patient (2.5%) reported severe pain when dressings were eliminated. On the first day during surgical cleaning procedure, 25% of cases reported severe pain and 25%, moderate pain; on the second day, 3.3% reported severe pain and 14.1% reported moderate pain. On the third day, 0.8% reported severe pain and 6.6% reported moderate pain; while on the fourth day, no one reported severe pain and only 12.5% reported moderate pain. On the fifth day, noone reported severe pain and only 0.8% reported moderate pain. DISCUSSION: The surgical cleansing methods applied in wounds originated in head and neck abscess debridement produce pain that could be controlled by irrigating lidocaine solution immediately before and during the process of cleansing and is an alternative method bacause general anesthesia, profound sedation, and direct anesthetic injection are contraindicated. This technique was effective in a limited fashion because multiples factors modified local analgesia. This technique should be used in decontamination because dressing materials are eliminated and before wound cleansing action begins. PMID- 14558969 TI - [Prognostic factors in oropharyngeal cancer]. AB - INTRODUCTION: Oropharyngeal carcinoma (OFC) is rare in Mexico, it only represents 0.6% of all carcinomas. Treatment alternatives are radiotherapy (RT) or radiotherapy-surgery association and prognosis depends on stage and tumor location. OBJECTIVE: Our objective was to know prognostic factors in patients with OFC treated with RT. MATERIAL AND METHODS: We conducted a retrospective analysis of patients with OFC. Analyzed variables were tumor location, RT technique (with or without field reduction; TRC and TSRC), tumor stage, histologic grade, and macroscopic tumor type. Survival was estimated with Kaplan Meier method. RESULTS: A total of 70 patients, median age of 62 years, 19 women and 51 men. Locations included 24 tonsil, 35 tongue base, eight soft palate, and three posterior lateral wall. A total of 32 tumors were excrescent and 38, ulcer infiltrating, 81% T3-T4 and 18% T1-T2. RT median dose was 5,625 rads; 51 were treated with TSRC and 19 with TRC. Local-regional control was achieved in 35 patients (50%), mortality secondary to treatment was 12.9% whit there were 5-year overall survival. Loco-regional recurrence was 44%. Most frequent morbility for treatment was dysphagia (66%). Variables with statistical survival significance were TRC (18% vs. 53% p = 0.0012), stage (T1-2 vs. T3-4; 58% vs. 21% p = 0.0025, and tonsil location (43% vs. 24% p < 0.005). OFC diagnosis is usually made in advanced stages. Survival prognostic factors are tumor location, tumor stage, and RT technique. PMID- 14558970 TI - [Myocardial revascularization in patients with ischemic heart disease and left ventricular dysfunction]. AB - BACKGROUND: Operative morbidity and mortality in coronary artery bypass surgery has decreased over the last decades and coronary artery bypass surgery offers an increase in long-term survival in selected patients with lower ejection fraction. We do not know exactly the results of the surgery and evolution of our patients with chronic lower ejection fraction. OBJECTIVE: To evaluate clinical results of myocardial revascularization in patients with ischemic heart disease and chronic lower ejection fraction. MATERIAL AND METHODS: We analyzed the experience between January 2000 and December 2001 of patients with ejection fraction = 30% treated with surgical myocardial revascularization. The procedure was carried out with cardiopulmonary bypass and moderate hypothermia (30 degrees C), and myocardial protection with intermittent cold crystalloid cardioplegia. We evaluated demographic characteristics, surgical procedure carried out, length-of-stay in intensive postoperative care unit, and morbidity and mortality in perioperative period and in middle term follow-up. RESULTS: During this period, 738 patients were treated with aorto-coronary bypass surgery Thirty eight patients were patients with chronic lower ejection fraction (5.14%) and were included in this evaluation. Six patients were female (15.78%), and 32 males (84.21%). Range of duration of cardiopulmonary bypass was 65-245 min (mean, 123 min), range of aortic cross-clamping time was 30-120 min (mean, 59 min). Twelve month follow-up showed improvement in two functional classes in 15 patients (39.4%), and improvement in one functional class in 10 patients (26.3%); two patients remained in the same functional class (5.2%), and one patient decreased his/her functional class (2.6%). Mean of length-of-stay in intensive care unit was 6.5 days and in hospital, 12.5 days. Perioperative morbility was 34.2% and same-period mortality was 26%; 12 month survival was 74%. It was concluded that even with high mortality as in other series, survival and quality of life of these patients is better with than medical therapy. PMID- 14558971 TI - [Duodenogastroesophageal reflux- and esophagitis-induced disease]. AB - OBJECTIVE: Our objective was to determine the relationship between reflux of duodenogastric contents with different degrees of esophagitis, and its levels and activity, with regard to the severity of esophagitis-induced duodenogastroesophageal reflux disease (DGERD). MATERIAL AND METHODS: Our study design was prospective, transversal, descriptive, observational, and open. We took samples of gastric and esophageal juices from patients with a diagnosis of gastroesophageal reflux disease (GERD) and esophagitis during the period from March to August 2002. Sample material was placed in black rubber-covered assay tubes to prevent bilirubin degradation. In our Unit's central laboratory, these samples were centrifuged for 30 min, the dissolved material separated, and a reactive strip was introduced to measure the amount of bile pigment. Endoscopic study reports were provided by this Department on terminating the procedure. Univariate analysis was used to obtain results. RESULTS: In 60% of cases, the esophageal liquid was positive for total bilirubin; 40% presented stage II esophagitis, 27% chronic esophagitis, and 27%, stage I esophagitis. Specific quantifications of total bilirubin at different stages of esophagitis demonstrated in stage II 50% with levels of 0.1-1, 33% with a level of 2-3, in cases of chronic esophagitis 75% with levels of 0.1-1, and with stage I esophagitis, 25% with levels of 2-3 mg/dl. Measurement of gastric pH showed 18 (69%) <3 and esophageal pH of 20 (67%) > 4, with 10 (33%) referring occasional or asymptomatic dyspepsia. We determined presence of duodenal elements in esophagus by means of quantification of total bilirubin in 60% of patients, observing greatest frequency of stage II esophagitis. Low but continuous levels of duodenal, principally biliary, elements continued in patients with DGERD, producing greatest damage in esophageal mucosa. PMID- 14558972 TI - [Hydropneumatic dilatation of the ureter: A technical option in ureteropyeloplasty]. AB - INTRODUCTION: The pyeloureteric junction (UP) is the site of obstruction commonest in the upper tract urinary. Several surgical techniques exist and the main complication is the stricture by the disproportion between both ends. The hydropneumatic dilatation of the ureteric would increase of the 5:1 to 2:1 the light of the ureter, improving technically anastomosis and reducing complications. MATERIAL AND METHODS: They studied 27 patients with diagnose of Estenosis UP in the Paediatric Urologist Service of CMN 20 of November of the ISSSTE, being made dismembered Pyeloplasty pyeloureteric in all, introducing Fogarty catheter in the Ureter carrying out the hydropneumatic dilatation of the same one, introduction of catheter double J in ureter and in the last patient without this, previous one to the anastomosis. All had a pursuit of 6 months. RESULTS: Of the 27 patients, 21 masculine ones (77.78%), 6 feminine ones (22.22%), of the right side 10 (37.04%), left 14 (51.85%) and bilateral 3 (11.11%). In the 27 studied patients postoperating complications did not appear. To all the patients I am made the dilatation to them hydropneumatics of the Ureter during UP plasty, did not present/display postoperating complications, the anastomosis is carried out with greater technical facility since the light of the Ureteric increase from 5:1 in 2:1, and in I complete case without ferulization of the Ureter, I do not present/display complications. DOSCUSSION: The hydropneumatics dilatation of the Ureter is a technical option in the Surgical handling of stricture UP, facilitating the anastomosis, diminishing the postoperating complications, making the ferulization do withoutible of the Ureter. PMID- 14558973 TI - [Usefulness of bispectral index in pharmaceutical cost reduction for anesthesia]. AB - BACKGROUND: The bispectral index (BIS) is a value derived from an electroencephalograph (EEG); it is correlated with depth of sedation and loss of consciousness. Therefore, it has been considered that its control on sedation depth could influence cost saving in drugs as well as decreased anesthesia costs. METHODS: A total of 175 patients were studied. One hundred patients were given intravenous (i.v.) anesthesia, 50 were observed with a BIS monitor, and the remainder went into the control group. Seventy five patients were given balanced general anesthesia: fifty were observed with BIS monitor, while the remainder functioned as the control group. Drug consumption per patient was measured a to maintain BIS value between 60 and 40, and the cost was calculated. RESULTS: Average drug costs for anesthesia were greater in BIS-controlled groups. Anesthesia cost/h was lower in prolonged procedures (>4 h). The bispectral Index as a sedation monitor during anesthesia is an excellent tool, although it did not show a real economic advantage, and we considered that it world only be feasible during long-term procedures. PMID- 14558974 TI - [Causality and hospital perinatal mortality trend at the Mexican Institute of Social Security, 1998-2002]. AB - OBJECTIVE: To analyze levels, trend and causes of hospital perinatal mortality at the Instituto Mexicano del Seguro Social (IMSS) during the 5 years from 1998 to 2002 to identify magnitude and related factors in our population and discuss some technical bases and epidemiologic aspects for planning strategies to contribute to its reduction. MATERIAL AND METHODS: Descriptive study on the death certificates of 39,994 cases of perinatal deaths distributed among 19,108 fetal deaths of 28 weeks and more of gestation and 20,886 neonatal deaths 7 to days of life that occurred in IMSS hospitals during the reference period. With this information and the data on total births, dead or live from the official information system of our Institution, we established annual rates for the entire IMSS, including administrative regions and zones of medical services. We also generated 5-year cumulated numbers for frequencies and rates of perinatal death causes in the IMSS, using the codes of Tenth Revision of the International Diseases Classification. RESULTS: Hospital perinatal mortality at the IMSS showed a reduction from 1998 (13 per 1,000 births) to 2002 (11.4 per 1,000 births). That trend was observed in the four administrative regions and in the majority of number of medical services zones, but with many differences in levels. It was similar in proportions of fetal (47.8%) and neonatal deaths (52.2%). Two thirds of fetal mortality was linked to maternal complications during pregnancy and labor. A similar proportion of neonatal deaths was due to premature birth and its complications. PMID- 14558975 TI - [Gallbladder ascariasis with acute hepatitis. Conservative treatment]. AB - We report two cases of gallbladder ascaridiasis associated with acute hepatitis, its clinical evolution with conservative treatment, making diagnosis by both laboratory and ultrasono-graphic studies. Case 1: was a male in his early forties who experienced symptoms of acute hepatitis and cholecystitis within a time lapse of 72 h of evolution. When laboratory tests and ultrasound (US) were done, an ascaris inside gallbladder was corroborated. There were also alterations compatible with acute non-viral hepatitis. Conservative treatment was done with observations within an 8-day period that hepatic examinations were normal as well as absence of helminthus inside gallbladder. Case 2: A 10-year-old female, who expelled worms 8 months previously had 11 days evidence of acute cholecystitis and hepatitis. An ultrasound of liver and biliary tract was done, with evidence of Ascaris lumbricoides inside gallbladder, with alterations in hepatic tests. This was medically treated, achieving expulsion of the Ascaris lumbricoides from inside the gallbladder and normalization of liver function tests. Gallbladder ascaridiasis management may be conservative. Patient general condition must be evaluated, as well or medical evolution and associated pathologies that may interfere in certain ways in surgery. Follow-up of these patients must be strict, with medical evaluation and laboratory controls. PMID- 14558976 TI - [Persistent left superior vena cava: infrequent localization of central venous catheter]. AB - OBJECTIVE: Our objective was to report a Persistent Left Superior Vena Cava in a critically ill patient. DESIGN: Case report. PLACE: Intensive care unit of a referral center. PATIENT: A 52 years-old male without previous cardiovascular disease was admitted to the intensive care unit due to cranial trauma. A central left subclavian vein catheter was placed at his arrival at the center chest X-ray showed the catheter in the left of the cardiac silhohuete without hemo pneumothorax. A angiographic evaluation through central venous catheter showed persistent left superior vena cava draining to the coronary sinus and right atrium with normal permeability of right superior vena cava. Other congenital alterations were excluded. Chest computed tomography confirmed the diagnosis. Persistent left superior vena cava is a congenital vascular abnormality. The intensive care unit staff must be aware of this vascular abnormality because it can complicate central venous catheterization. PMID- 14558977 TI - [The role of minimal invasion surgery in the management of patients with gastric carcinoma]. AB - In the last decade, minimal invasion surgery (MIS) has been used in a great variety of diseases and clinical situations. In gastric carcinoma (GC), specific indications in various circumstances have been described. MIS is used to make a diagnosis, for clinical stage classification, and for respectability evaluation, allowing to design a suitable therapy for each patient, because effective neoadjuvant treatments (chemotherapy and radiochemotherapy) are available. MIS has also been used for surgical resection of gastric neoplasia, including endoscopic mucosal resection, wedge resection of gastric wall, subtotal gastrectomy, total gastrectomy or even lymphadenectomy. MIS is used for perform palliative gastrojejunostomy in patients with distal GC, as well as for gastrostomy or jejunostomy for enteral nutrition in specific situations. MIS is still in experimental phase in GC, except in the case of preoperative staging laparoscopy. PMID- 14558978 TI - [Perioperative lesions in lower extremity peripheral nerves]. AB - OBJECTIVE: To review the actual concepts concerning perioperative nerve injuries during surgical or anesthetic procedures. MATERIAL AND METHODS: A summary of published medical literature from Medline search files and published reviews. CONFLICT IF INTEREST: We have not conflicts of interest. RESULTS: Perioperative nerve injuries during surgical and anesthetic procedures of the lower extremities are the most common. Many possible etiologies have been proposed to explain perioperative nerve injury and include stretch, compression, and ischemia. Commonly this injuries are under recognition and an important aspect of perioperative nerve injuries is that they can occur for multiple reasons. It is necessary to have a good communication among surgeons, neurologists and clinical neurophysiologists, so it can lead to a more detailed neurological assessment with transoperative procedures that may prevent such injuries. PMID- 14558979 TI - [Commentary on the article entitled: "Association of physiologic responses to metabolic changes in extenuating physical exercise"]. PMID- 14558980 TI - [Two key words, Cirugia y Cirujanos, their anatomy]. PMID- 14558981 TI - Do selective cyclo-oxygenase-2 nonsteroidal anti-inflammatory agents increase the risk for acute myocardial infarction? PMID- 14558982 TI - Trials in isolated systolic hypertension: an update. AB - Isolated systolic hypertension (ISH) has proved to be a powerful predisposing factor for cardiovascular diseases in the elderly. Recent placebo-controlled interventional trials such as the Systolic Hypertension in the Elderly Program, the Systolic Hypertension in Europe, and the Systolic Hypertension in China showed that the lowering of systolic blood pressure using a diuretic- or a calcium antagonist-based treatment is associated with a decrease in cardiovascular events. Antihypertensive therapy was found especially effective in preventing stroke in the elderly with ISH. A slowing in the progression of dementia was observed in patients randomized to a calcium antagonist-based treatment. Patients at high cardiovascular risk such as those with diabetes benefited the most from treatment. In another trial performed in patients with left ventricular hypertrophy (Losartan Intervention For Endpoint Reduction), a subset of patients had ISH. In those patients, an angiotensin II antagonist-based treatment was superior to a b-blocker-based treatment in preventing cardiovascular complications. The experience accumulated in patients with ISH showed that combination therapy is often required to control blood pressure. Overall, the evidence available today indicates that pharmacologic treatment of ISH markedly improves the outcome of elderly patients. PMID- 14558983 TI - Hypertension and dementia. AB - Hypertension is one of the principal risk factors for cerebrovascular diseases. Several epidemiologic studies have also indicated a positive correlation between cognitive decline or dementia and blood pressure level. Indeed, the results of most longitudinal studies show that cognitive functioning is often inversely proportional to blood pressure values measured 15 or 20 years previously. Cerebral infarcts, lacunae, and white matter changes are implicated in the pathogenesis of vascular dementia, but may also favor the development of Alzheimer's disease. Microcirculation disorders and endothelial dysfunctions are also advanced to explain the deterioration in cognitive functions in hypertensive subjects. Data from recent therapeutic trials open the way to the prevention of dementia (vascular or Alzheimer's type) by antihypertensive treatments and must be confirmed by other studies. PMID- 14558984 TI - The J-curve in hypertension. AB - The J-curve debate has continued for 25 years. Dependency upon observational and retrospective studies has confused the issue; only the full publication of data from the prospective, randomized Hypertension Optimal Treatment (HOT) study has thrown genuine light on the problem. Many examples of the J-curve relationship between blood pressure and cardiovascular/noncardiovascular events are due to reverse causality, where underlying disease (eg, poor left ventricular function, poor general health, poorly compliant/stiff arteries) is the cause of both the low blood pressure and the increased risk of both cardiovascular and noncardiovascular events. The J-curve in patients with stiff arteries (wide pulse pressure) may be exacerbated by treatment. From the full publication of the HOT study database it is now reasonable to conclude that for nonischemic hypertensive subjects the therapeutic lowering of diastolic blood pressure (DBP) to the low 80s mm Hg is beneficial, but it is safe (though unproductive) to go lower. However, in the presence of coronary artery disease (limited coronary flow reserve) there is a J-curve relationship between treated DBP and myocardial infarction, but not for stroke. In such high-risk (for myocardial infarction) cases it would be prudent to avoid lowering DBP to below the low 80s mm Hg. PMID- 14558985 TI - Leptin: sympathetic and cardiovascular effects. AB - Shortly after leptin was first discovered, it was hailed as the key to understanding obesity. However, it didn't take long for investigators to realize that the hormone was more than a feedback signal to inhibit further food intake. Since those early days, leptin has been well characterized in rodents. It exerts an influence in many physiologic processes, including food intake, thermoregulation, fertility, thyroid function, adrenal function, sympathetic nerve activation, renal function, blood vessel tone, and blood pressure. No longer a satiety hormone, it is being looked at from many different perspectives. One such perspective is its influence on the cardiovascular system. This review highlights some of the work in this area. PMID- 14558988 TI - Newer pharmaceutical agents to treat lipid disorders. AB - The National Cholesterol Education Program Adult Treatment Panel III guidelines and the results of the Heart Protection Study have provided a stronger rationale to more aggressively treat high-risk patients to a low-density (LDL) cholesterol goal of less than 100 mL/dL. Two new therapies, ezetimibe and rosuvastatin, have recently been added to the lipid-lowering armamentarium to improve guideline adherence. Ezetimibe, a novel cholesterol absorption inhibitor, lowers LDL by 18% to 20% and can be used safely in combination with statins. Adding ezetimibe to a statin is comparable with the LDL-lowering efficacy of tripling the dose of the statin. Rosuvastatin is a highly efficacious statin providing 8% greater LDL reduction than equivalent doses of atorvastatin, and the starting dose of 10 mg/d provides nearly a 50% reduction in LDL cholesterol. There are several investigational drugs in development for the prevention and treatment of atherosclerosis. Of these investigational drugs, the most promising are the cholesterol ester transfer protein inhibitors, which have the potential to significantly raise high-density lipoprotein cholesterol and acetyl-coenzyme A: cholesterol acyltransferase inhibitors, which may directly inhibit the progression of atherosclerosis. PMID- 14558990 TI - Estrogen therapies, lipids, and the heart disease prevention controversy. AB - One of the most highly unexpected reports in recent medical literature was the lack of benefit of estrogen-progestin replacement therapy in cardiovascular disease prevention in postmenopausal women. The ensuing negative view of hormone replacement therapy has now extended to all forms of postmenopausal hormone treatment, including estrogen alone. Is this pessimism justified? A review of the effects of estrogens and progestins on the estrogen-sensitive systems of the body can help explain why combined oral estrogen and low-dose continuous medroxyprogesterone acetate administration may not be the paradigm for all other forms of postmenopausal hormone replacement. Some of these effects include the following: progestins are anti-estrogens, as evidenced in their divergent effects on plasma lipids; not all progestins are equal in their effect on lipids and other physiologic functions; administration of any hormone by mouth is not physiologic; giving estrogen 10 to 15 years postmenopausally may be too late to prevent atherosclerosis. On the other hand, high doses of oral estrogen/progestin in the presence of high cardiovascular risk appear to promote atherosclerosis risk. Given the current evidence the common sense answer to the question of the benefit of estrogen is "it depends." Until these and other points are formally addressed, the hypothesis that estrogen prevents heart disease remains open. PMID- 14558989 TI - Niacin, lipids, and heart disease. AB - Niacin is the most effective medication in current clinical use for increasing high-density lipoprotein (HDL) cholesterol. It has the broadest effect on the lipid profile, reducing all atherogenic apolipoprotein (apo) B and increasing all antiatherogenic apo AI-containing lipoproteins, resulting in significant reduction in atherosclerotic complications and total mortality in trials. Recent research indicates novel major target sites of action in the liver to 1) directly inhibit diacylglycerol acyltransferase 2 (DGAT2), explaining its effect on triglycerides and apo B lipoproteins, and 2) inhibit the HDL apo AI catabolism pathway, resulting in higher HDL levels. Such information may lead to new drug discovery and supply the rationale for combination with other lipid regulators that are known to have different mechanisms of action. Trial evidence shows that niacin is not only safe to use in persons with diabetes, but that its combination with 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG CoA) reductase inhibitors (statins) is also safe and effective. Recently, a new formulation of niacin has made it easier to tolerate and administer. Clinical trials are needed to determine whether niacin in combination with other lipid-modulating agents decreases the risk of cardiovascular events beyond the approximately 30% that has been noted with monotherapy. PMID- 14558991 TI - Plasma lipoprotein concentrations in ethnic populations. AB - There is a complex interplay between genetic and environmental factors that influences the expression of plasma lipoprotein levels. It is therefore not surprising that differences in lipid levels have been reported between ethnic groups. There are conflicting data on racial and ethnic variations in lipids, and also limited data on the relationship between lipoprotein levels and coronary heart disease risk in specific populations. This review summarizes available data on ethnic variations in plasma lipoproteins and the potential impact on coronary morbidity and mortality. PMID- 14558993 TI - DNA microarray for discrimination between pathogenic 0157:H7 EDL933 and non pathogenic Escherichia coli strains. AB - The primary technique currently used to detect biological agents is based on immunoassays. Although sensitive and specific, currently employed immunoassays generally rely on the detection of a single epitope, and therefore often cannot discriminate subtle strain-specific differences. Since DNA microarrays can hybridize hundreds to thousands of genomic targets simultaneously and do not rely on phenotypic expression of these genetic features for identification purposes, they have enormous potential to provide inexpensive, flexible and specific strain specific detection and identification of pathogens. In this study, pathogenic Escherichia coli O157:H7-specific genes, non-pathogenic K12-specific genes, common E. coli genes, and negative control genes were polymerase chain reaction amplified and spotted onto the surface of treated glass slides. After labeled bacterial cDNA samples were hybridized with probes on the microarray, specific fluorescence patterns were obtained, enabling identification of pathogenic E. coli O157:H7 and non-pathogenic E. coli K12. To test the utility of this microarray device to detect genetically engineered bacteria, E. coli BL21 (a B strain derivative with antibiotic resistance gene, ampR) and E. coli JM107 (a K12 strain derivative lacking the gene ompT) were also employed. The array successfully confirmed the strain genotypes and demonstrated that antibiotic resistance can also be detected. The ability to assess multiple data points makes this array method more efficient and accurate than a typical immunoassay, which detects a single protein product. PMID- 14558992 TI - Physical activity and its effects on lipids. AB - Physical activity is an important component of weight control, and is widely recommended to prevent and treat obesity-related complications such as diabetes and coronary heart disease (CHD). Although the cardiovascular benefits of increased physical activity are likely multifactorial, much of the attention has been focused on the known high-density lipoprotein (HDL) cholesterol-raising properties of regular physical activity. Physical activity, however, can also reliably lower triglycerides and favorably affect both low-density lipoprotein (LDL) and HDL particle sizes. Limited data on resistance exercise suggest that this type of physical activity may reduce LDL cholesterol. Although these lipid effects are modest and variable, they are likely to be particularly important in reducing the morbidity and mortality from CHD on a population level, and may be especially important in patients with atherogenic dyslipidemia. PMID- 14558994 TI - Quartz crystal biosensor for real-time monitoring of molecular recognition between protein and small molecular medicinal agents. AB - A quartz crystal microbalance (QCM) biosensor integrated into a flow injection analysis (FIA) system was used for the real-time investigation of molecular recognition between a protein and small molecular medicinal agents. Two sulfa drugs, sulfamethazine (SMZ) and sulfamethoxazole (SMO), were, respectively, immobilized on the gold electrodes of the piezoelectric crystals using appropriate procedures based on self-assembly of the dithiothreitol (DTT). The binding interactions of the two immobilized drug ligands, with various proteins in solution, were followed as changes in the resonant frequency of the modified crystals. Results obtained from this rapid screen analysis clearly indicated that the two drug ligands appeared quite different in this molecular recognition procedure although their structures were similar. SMZ-immobilized sensor showed specific interaction only with IgG, while SMO-immobilized sensor showed negligible specific binding with IgG, but binding with trypsin and chymotrypsin. Further studies on the specific interaction between immobilized SMZ and three different species of IgG--human IgG, goat IgG and mouse IgG were carried out and the marked species-dependent difference was observed. The resultant sensorgrams were rapidly analyzed by using an in-house kinetic analysis software based on genetic algorithm (GA) to derive both the kinetic rate constants (kass and kdiss) and equilibrium association constants (KA) for IgG-SMZ interactions. For the interactions, KA were 5.48 x 10(5), 2.75 x 10(5) and 1.86 x 10(5) M(-1) for human IgG, goat IgG and mouse IgG, respectively. The kinetic data provided further insight into the structural/functional relationships of different IgG on a molecular level. PMID- 14558995 TI - Microfluidic enzyme immunosensors with immobilised protein A and G using chemiluminescence detection. AB - Affinity proteins were covalently immobilised on silicon microchips with overall dimensions of 13.1 x 3.2 mm, comprising 42 porous flow channels of 235 microm depth and 25 microm width, and used to develop microfluidic immunosensors based on horseradish peroxidase (HRP), catalysing the chemiluminescent oxidation of luminol/p-iodophenol (PIP). Different hydrophilic polymers with long flexible chains (polyethylenimine (PEI), dextran (DEX), polyvinyl alcohol, aminodextran) and 3-aminopropyltriethoxysilane (APTS) were employed for modification of the silica surfaces followed by attachment of protein A or G. The resulting immunosensors were compared in an affinity capture assay format, where the competition between the labelled antigen and the analyte for antibody-binding sites took place in the bulk of the solution. The formed immunocomplexes were then trapped by the microchip affinity capture support and the amount of bound tracer was monitored by injection of luminol, PIP and H2O2. All immunosensors were capable of detecting atrazine at the sub-microg l(-1) level. The most sensitive assays were obtained with PEI and DEX polymer modified supports and immobilised protein G, with limits of detection of 0.006 and 0.010 microg l(-1), and IC50 values of 0.096 and 0.130 microg l(-1), respectively. The protein G based immunosensors were regenerated with 0.4 M glycine-HCl buffer pH 2.2, with no loss of activity observed for a storage and operating period of over 8 months. To estimate the applicability of the immunosensors to the analysis of real samples, PEI and DEX based protein G microchips were used to detect atrazine in surface water and fruit juice, spiked with known amounts of the atrazine, giving recovery values of 87-102 and 88-124% at atrazine fortification levels of 0.5-3 and 80-240 microg l(-1), respectively. PMID- 14558996 TI - Microplate based biosensing with a computer screen aided technique. AB - Melanophores, dark pigment cells from the frog Xenopus laevis, have the ability to change light absorbance upon stimulation by different biological agents. Hormone exposure (e.g. melatonin or alpha-melanocyte stimulating hormone) has been used here as a reversible stimulus to test a new compact microplate reading platform. As an application, the detection of the asthma drug formoterol in blood plasma samples is demonstrated. The present system utilizes a computer screen as a (programmable) large area light source, and a standard web camera as recording media enabling even kinetic microplate reading with a versatile and broadly available platform, which suffices to evaluate numerous bioassays. Especially in the context of point of care testing or self testing applications these possibilities become advantageous compared with highly dedicated comparatively expensive commercial systems. PMID- 14558997 TI - A high-performance glucose biosensor based on monomolecular layer of glucose oxidase covalently immobilised on indium-tin oxide surface. AB - In this paper, a mediatorless amperometric glucose biosensor based on direct covalent immobilisation of monomolecular layer of glucose oxidase (GOx) on a semiconducting indium-tin oxide (ITO) is demonstrated. The abundance of surface hydroxyl functional group of ITO allows it to be used as a suitable platform for direct covalent immobilisation of the enzyme for sensor architecture. The anodic current corresponding to electrochemical oxidation of the enzymatic product, hydrogen peroxide, at a sputtered Pt electrode at 0.500 V (vs. SCE) was obtained as the sensor signal. It was found that the biosensor based on the direct immobilisation scheme shows a fast biosensor response, minimum interference from other common metabolic species and ease of biosensor miniaturisation. A linear range of 0-10 mM of glucose was demonstrated, which exhibits a high sensitivity as far as performance per immobilised GOx molecule is concerned. A detection limit as low as 0.05 mM and long-term stability were observed. Even more important, the biosensor design allows fabrication through a dry process. These characteristics make it possible to achieve mass production of biosensor compatible with the current electronic integrated circuit manufacturing technologies. PMID- 14558998 TI - Biocompatibility of a physiological pressure sensor. AB - A newly developed fiber optic micropressure sensor was evaluated for biocompatibility using the International Organization for Standardization (ISO) test standard 10993-6. The test material and an inert control (fused silica glass) were tested in New Zealand white rabbits. Four test specimens were implanted in the paravertebral muscles on one side of the spine about 2-5 cm from the mid-line and parallel to the spinal column. Similarly, four control specimens were implanted on the opposite side. The implantation periods were 1, 4, and 12 weeks to ensure a steady state biological tissue response. Four animals were tested at each time period. Macroscopic and microscopic observations were performed to compare the biological reactions between the test and control materials. There was an inflammatory reaction at 1 week which subsided at 4 weeks. There was fibrous tissue growth near the implant that also decreased over time. Most importantly, there was no significant difference in the biological response between the test and control materials. Therefore, we conclude that the pressure microsensor is biocompatible. PMID- 14558999 TI - Nucleic acid purification using microfabricated silicon structures. AB - A microfluidic device has been designed, fabricated and tested for its ability to purify bacteriophage lambda DNA and bacterial chromosomal DNA, a necessary prerequisite for its incorporation into a biosensor. This device consists of a microfabricated channel in which silica-coated pillars were etched to increase the surface area within the channel by 300-600%, when the etch depth is varied from 20 to 50 microm. DNA was selectively bound to these pillars in the presence of the chaotropic salt guanidinium isothiocyanate, followed by washing with ethanol and elution with low-ionic strength buffer. Positive pressure was used to move solutions through the device, removing the need for centrifugation steps. The binding capacity for DNA in the device was approximately 82 ng/cm2 and on average, 10% of the bound DNA could be purified and recovered in the first 50 microl of elution buffer. Additionally, the device removed approximately 87% of the protein from a cell lysate. Nucleic acids recovered from the device were efficiently amplified by the polymerase chain reaction suggesting the utility of these components in an integrated, DNA amplification-based biosensor. The miniaturized format of this purification device, along with its excellent purification characteristics make it an ideal component for nucleic acid-based biosensors, especially those in which nucleic acid amplification is a critical step. PMID- 14559001 TI - Development of a conductimetric biosensor using immobilised Rhodococcus ruber whole cells for the detection and quantification of acrylonitrile. AB - A conductimetric biosensor for the detection of acrylonitrile in solution was designed and characterised using whole cells of Rhodococcus ruber NCIMB 40757, which were immobilised into a disc of dimethyl silicone sponge (ImmobaSil). The biosensor described was capable of the detection and quantification of acrylonitrile in aqueous solution, having a linear response to concentrations between 2 and 50 mM (106-2650 ppm) acrylonitrile. The biosensor has been shown to be reproducible with respect to the data obtained over a number of days, and retains stability for a minimum period of at least 5 days before recalibration of the biosensor is required. PMID- 14559000 TI - Optical-transparent and flexible glucose sensor with ITO electrode. AB - The glucose sensor was constructed by immobilizing glucose oxidase (GOD) with glutaraldehyde solution onto the sensitive area of the transparent oxygen electrode. The oxygen electrode was fabricated by sealing KCl electrolyte solution including the Indium-Tin Oxide (ITO)-electrode with both metal-weldable film and gas-permeable membrane coated with Ag/AgCl electrode. The sensor behavior was evaluated using standard glucose solutions in a batch measurement system with a computer-controlled potentiostat at a reduction potential of -900 mV. The sensor device has flexible structure and good optical transparency (less than 0.6 abs) at the visible wavelength from 400 to 700 nm. The sensor was possible to be used for measuring glucose from 0.06 to 1.24 mmol/l (correlation coefficient: 0.999), including the reported concentration of tear glucose in normal (0.14 mmol/l), with good reproducibility. PMID- 14559002 TI - Galactosylated PVDF membrane promotes hepatocyte attachment and functional maintenance. AB - One of the major challenges in BLAD design is to develop functional substrates suitable for hepatocyte attachment and functional maintenance. In the present study, we designed a poly(vinylidene difluoride) (PVDF) surface coated with galactose-tethered Pluronic polymer. The galactose-derived Pluronic F68 (F68-Gal) was adsorbed on PVDF membrane through hydrophobic-hydrophobic interaction between PVDF and the polypropylene oxide segment in Pluronic. The galactose density on the modified PVDF surface increased with the concentration of the F68-Gal solution, reaching 15.4 nmol galactosyl groups per cm2 when a 1 mg/ml of F68-Gal solution was used. The adsorbed F68-Gal remained relatively stable in culture medium. Rat hepatocytes attachment efficiency on F68-Gal modified PVDF membrane was similar to that on collagen-coated surface. The attached hepatocytes on PVDF/F68-Gal membrane self-assembled into multi-cellular spheroids after 1 day of culture. These attached hepatocytes in spheroids exhibited higher cell functions such as albumin synthesis and P450 1A1 detoxification function compared to unmodified PVDF membrane and collagen-coated surface. These results suggest the potential of this galactose-immobilized PVDF membrane as a suitable substrate for hepatocyte culture. PMID- 14559003 TI - Craniofacial osteoblast responses to polycaprolactone produced using a novel boron polymerisation technique and potassium fluoride post-treatment. AB - There is no ideal material for craniofacial bone repair at present. The aim of this study was to test the biocompatibility of polycaprolactone (PCL) synthesised by a novel method allowing control of molecular weight and degradation rate, with regard to it being used as matrix for a biodegradable composite for craniofacial bone repair. Human primary craniofacial cells were used, isolated from paediatric skull after surgery. Cell responses were analysed using various assays and antibody staining. Cells attached and spread on the PCL in a similar manner to the Thermanox controls as shown by phalloidin staining of F-actin. Cells maintained the osteoblast phenotype as demonstrated by alkaline phosphatase assay and antibody staining throughout the time points studied, up to 28 days. Cells proliferated on the PCL as shown by a DNA assay. Collagen-1 staining showed extensive production of a collagen-1 containing extracellular matrix, which was also shown to be mineralised by alizarin red staining. Short-term (up to 48 h) attachment studies and long-term (up to 28 days) expression of markers of the osteoblast phenotype have been demonstrated on the PCL. This new method of synthesising PCL shows biocompatibility characteristics that give it potential to be used for craniofacial bone repair. PMID- 14559004 TI - Action of FGMgCO3Ap-collagen composite in promoting bone formation. AB - To improve the biological properties of materials as bone substitutes, functionally graded CO3 apatite crystals containing magnesium, FGMgCO3Ap, were synthesized to be mixed with atelocollagen and made into a composite pellet. A radio-labeled cell adhesion experiment showed that the degree of adherence of mouse MC3T3E1 osteoblast-like cells to the FGMgCO3Ap-collagen composite was better than to CO3Ap-collagen and much better than to the Ti plate. When the composites were implanted beneath the periosteum cranii of rats, the FGMgCO3Ap collagen composite was metabolized faster than the CO3Ap-collagen composite and better formation of new bone and osteoblast arrangement at the interface between the composite and the periosteum cranii was observed. When the composites were implanted into the femur of rabbits, clear bone formation with a higher degree of bone density was observed for the FGMgCO3Ap-collagen composite. These results suggest that the Mg2+ ions taken into the apatite crystals may contribute to the acceleration of osteoblast adhesion to apatites and promote bone formation, cross talking with osteoblasts at the molecular level. PMID- 14559005 TI - Preparation and antibacterial effects of Ag-SiO2 thin films by sol-gel method. AB - In the present study, silver-doped silica thin films were successfully prepared by sol-gel method to apply for antibacterial materials. The starting solution was prepared from 1:0.24:3.75:2.2 molar ratios of Si(OC2H5)4):AgNO3:H2O:C2H5OCH2CH2OH and then the pH value controlled at 3 with 0.5 N HNO3 solution. The formation of silver-doped glassy silica thin films at various temperatures was investigated through infrared spectroscopy, ultraviolet-visible, scanning electron microscopy and X-ray diffraction. From these analysis data, it was found that silver ions were completely trapped in the silica matrix and their reduction could be achieved at 600 degrees C annealing temperature. The antibacterial effects of silica thin films against Escherichia coli and Staphylococcus aureus were examined by film attachment method. The coating films had an excellent antibacterial performance. PMID- 14559006 TI - A new austenitic stainless steel with negligible nickel content: an in vitro and in vivo comparative investigation. AB - New nickel (Ni)-reduced stainless-steel metals have recently been developed to avoid sensitivity to Ni. In the present study, an austenitic Ni-reduced SSt named P558 (P558, Bohler, Milan, Italy) was studied in vitro on primary osteoblasts and in vivo after bone implantation in the sheep tibia, and was compared to ISO 5832 9 SSt (SSt) and Ti6Al4V. Cells were cultured directly on P558 and Ti6Al4V. Cells cultured on polystyrene were used as controls. Osteoblast proliferation, viability and synthetic activity were evaluated at 72 h by assaying WST1, alkaline phosphatase activity (ALP), nitric oxide, pro-collagen I (PICP), osteocalcin (OC), transforming growth factor-beta1 (TGFbeta-1) and interleukin-6 (IL-6) after 1.25(OH)2D3 stimulation. Under general anaesthesia, four sheep were submitted for bilateral tibial implantation of P558, SSt and Ti6Al4V rods. In vitro results demonstrated that the effect of P558 on osteoblast viability, PICP, TGF beta-1, tumor necrosis factor-alpha production did not significantly differ from that exerted by Ti6Al4V and controls. Furthermore, P558 enhanced osteoblast differentiation, as confirmed by ALP and OC levels, and reduced IL-6 production. At 26 weeks, the bone-to-implant contact was higher in P558 than in SSt (28%, p<0.005) and Ti6Al4V (4%, p<0.05), and was higher in Ti6Al4V than in SSt (22%, p<0.005). The tested materials did not affect bone microhardness in pre-existing host bone as evidenced by the measurements taken at 1000 microm from the bone biomaterial interface (F=1.89, ns). At the bone-biomaterial interface the lowest HV value was found for SSt, whereas no differences in HV were observed between materials (F=1.55, ns). The current findings demonstrate P558 biocompatibility both in vitro and in vivo, and osteointegration processes are shown to be significantly improved by P558 as compared to the other materials tested. PMID- 14559007 TI - NMR properties of alginate microbeads. AB - Alginates are a family of unbranched polysaccharides with properties that vary widely depending on their composition. In the presence of multivalent cations (frequently Ca2+), alginates form a gel. Consequently, alginates have been used to encapsulate a variety of biological materials, including cells. In this study, we present NMR relaxation and diffusion data from alginate microbeads with similar size and properties to those used in the development of a bioartificial pancreas. Our data demonstrate that the transverse relaxation time (T2) of water within the gel depends on the guluronic acid content of the alginate, whereas the longitudinal relaxation time (T1) and the apparent diffusion coefficient of water do not. Our data further suggest that the diffusion of Ca2+ ions is hindered by the presence of a poly-L-lysine layer, a layer commonly added to provide mechanical support to the beads and immunoprotection to the encapsulated cells in the event of implantation. The impact of these data on our understanding of the role of alginate gels in the development of a bioartificial pancreas is discussed. PMID- 14559008 TI - Peptide functionalized poly(L-lysine)-g-poly(ethylene glycol) on titanium: resistance to protein adsorption in full heparinized human blood plasma. AB - The graft copolymer poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG) and its RGD- and RDG-functionalized derivatives (PLL-g-PEG/PEG-peptide) were assembled from aqueous solutions on titanium (oxide) surfaces. The polymers were characterized by NMR in order to determine quantitatively the grafting ratio, g (Lys monomer units/PEG side chains), and the fraction of the PEG side chains carrying the terminal peptide group. The titanium surfaces modified with the polymeric monomolecular adlayers were exposed to full heparinized blood plasma. The adsorbed masses were measured by in situ ellipsometry. The different PLL-g PEG-coated surfaces showed, within the detection limit of the ellipsometric technique, no statistically significant protein adsorption during exposure to plasma for 30 min at 22 degrees C or 37 degrees C, whereas clean, uncoated titanium surfaces adsorbed approximately 350 ng/cm2 of plasma proteins. The high degree of resistance of the PEGylated surface to non-specific adsorption makes peptide-modified PLL-g-PEG a useful candidate for the surface modification of biomedical devices such as implants that are capable of eliciting specific interactions with integrin-type cell receptors even in the presence of full blood plasma. The results refer to short-term blood plasma exposure that cannot be extrapolated a priori to long-term clinical performance. PMID- 14559009 TI - Histological and mechanical investigation of the bone-bonding ability of anodically oxidized titanium in rabbits. AB - The purpose of this study was to histologically and mechanically investigate the in vivo bone-bonding ability of anodically oxidized titanium (AO Ti) with an anatase crystal layer on its surface. AO Ti plates, anodically oxidized at 155 V in 1 M H2SO4, were implanted into the proximal metaphyses of mature rabbit tibiae for 4, 8, 16, and 24 weeks and investigated by light microscopy, scanning electron microscopy and detaching test. High bone-bonding ability, comparable to our previous study data of the bioactive titanium produced by sodium-free alkali and heat treatment, was observed at the early stages of implantation. However, no substantial increase was demonstrated. AO Ti plates bonded to bone directly, with no intervening soft tissue layer, and no breakage of the AO Ti layer was observed. The AO Ti layer was porous through to the titanium substrate, while the porosity was low. Apatite-like deposition into the pores of the AO layer was observed only in the superficial zone. The lack of improvement of bone-bonding ability in the later stages of implantation may be attributed to the low porosity and to the superficial ingrowths of apatite-like deposits into the pores of the AO Ti layer. PMID- 14559010 TI - Quantitative analysis of the resorption and osteoconduction process of a calcium phosphate cement and its mechanical effect for screw fixation. AB - The clinical application of calcium phosphate cements (CPCs) composed of tetracalcium phosphate and dicalcium phosphate anhydrous has been limited because of its longer setting time, so that we developed the CPC in which the setting time was shortened to approximately 10 min. Aiming at clinical application, we evaluated the histological response in the bone quantitatively and the biomechanical effectiveness of this substance. The CPC was implanted in the rabbit femoral condyle up to 52 weeks for histological evaluation. In mechanical testing, small cancellous screws were inserted into the condyle, both with and without augmentation with the CPC, and the pull-out strength was measured. The micro-computed tomography finding demonstrated that the cross-sectional area of the implanted CPC at 24 weeks was approximately two-thirds of the initial area. The amount of newly calcified bone around the CPC was significantly greater than that of the sintered hydroxyapatite. Histologically, the new bone was formed on the surface of the implanted CPC 1 week after the implantation and resorption of the CPC was evident at 3 weeks. The pull-out strength was enhanced significantly by augmentation with the CPC and the initial strength was maintained for a 6 week period. This CPC showed good osteoconductivity and was resorbed without adverse inflammation. Using the CPC as augmentation may be capable of useful treatment options in fractures with poor bone quality. PMID- 14559011 TI - Control of degradation rate and hydrophilicity in electrospun non-woven poly(D,L lactide) nanofiber scaffolds for biomedical applications. AB - Typical properties of poly(D,L-lactide) (PLA)-based scaffolds (films and foams), such as long degradation time, mechanical stiffness and hydrophobicity, are sometimes not suitable for biomedical applications. These properties can be substantially altered by electrospinning of PLA blends with miscible poly(lactide co-glycolide) (PLGA) random copolymers, poly(lactide-b-ethylene glycol-b-lactide) (PLA-b-PEG-b-PLA) triblock copolymers, and a lactide (used as a hydrolytic catalyst). Electrospun scaffolds based on the multi-component PLA blends, comprised of randomly interconnected webs of sub-micron sized fibers, have a bulk density of 0.3-0.4 g/cm3. In this study, the concentration effects of PLA-b-PEG-b PLA triblock copolymer and lactide on the cell proliferation and the hydrophilicity of electrospun scaffolds were investigated. Based on in vitro degradation study, we found that the electrospun scaffold having PLA (40 wt%), PLGA (LA/GA=50/50, 25 wt%), PLA-b-PEG-b-PLA (20 wt%), and lactide (15 wt%) underwent a rapid weight loss of approximately 65% in 7 weeks. The hydrophobicity of this membrane, as determined by contact angle measurements in a cell buffer solution, decreased by approximately 50% from 105 degrees (of an electrospun PLA scaffold) to 50 degrees. The selection of suitable chemical compositions in conjunction with the non-invasive electrospinning process is useful in the production of a new kind of biodegradable scaffolds suitable for different biomedical applications such as cell storage and delivery as well as prevention of post-surgical adhesion because of their porosity, mechanical flexibility and tunable biodegradability. PMID- 14559012 TI - Characterization of a bovine collagen-hydroxyapatite composite scaffold for bone tissue engineering. AB - Different biomaterials have been used as scaffolds for bone tissue engineering. Here we characterize a biomaterial composed of sintered (1100 degrees C) and powdered hydroxyapatite (HA) and type I collagen (Coll), both of bovine origin, designed for osteoconductive and osteoinductive scaffolds. Coll/HA proportions were 1/2.6 and 1/1 (wet weight), and particles sizes varied from 200 to 400 microm. Vv (volume density) and Sv (surface to volume density) for the HA particles in the composite ranged from 0.48 +/- 0.06 to 0.55 +/- 0.02 and 5.090 +/- 0.545 to 6.366 +/- 0.289 microm(-1), respectively. Due to the relatively small changes in Vv and Sv, a macroporosity could be characterized for the biocomposite. X-ray diffraction and infrared spectroscopy showed that the sintered bone was composed essentially of HA with minimum additional groups such as surface calcium hydroxide, surface and crystal water, free carbon dioxide and possibly brushite. Mass spectrometry detected carbonates at A and B sites of HA, and weakly bound to the structure. Human osteoblasts adhered and spread on both the HA particle surface and the collagen fibers, which seemed to guide cells between adjacent particles. The biocomposite studied has several characteristics considered as ideal for its use as a scaffold for osteoconduction and osteoinduction. PMID- 14559013 TI - Bone generation on PHBV matrices: an in vitro study. AB - Bone formation was investigated in vitro by culturing rat marrow stromal osteoblasts in biodegradable, macroporous poly(3-hydroxybutyric acid-co-3 hydroxyvaleric acid) (PHBV) matrices over a period of 60 days. Foams were prepared after solvent evaporation and solute leaching. PHBV solutions with different concentrations were prepared in chloroform: dichloromethane (1:2, v/v). In order to create a matrix with high porosity and uniform pore sizes, sieved sucrose crystals (300-500 microm) were used. PHBV foams were treated with rf oxygen plasma (100 W 10 min) to modify their surface chemistry and hydrophilicity with the aim of increasing the reattachment of osteoblasts. Osteoblasts were isolated from rat bone marrow and seeded onto PHBV foams. The cell density on and in the foams was determined with MTS assay. MTS results showed that osteoblasts proliferated on PHBV. Twenty-one days after seeding of incubation, growth of osteoblasts on matrices and initiation of mineralization were observed by confocal laser scanning microscopy. Increasing ALP and osteocalcin secretion during 60 days confirmed the osteoblastic phenotype of the derived stromal cells. SEM, histological evaluations and confocal laser scanning microscopy showed that osteoblasts could grow inside the matrices and lead to mineralization. Cells exhibited spindle-like morphology and had a diameter of 10-30 microm. Based on these, it could confidently be stated that PHBV seems to be a promising polymeric matrix material for bone tissue engineering. PMID- 14559014 TI - Pulsatile perfusion and cardiomyocyte viability in a solid three-dimensional matrix. AB - BACKGROUND: The manufacture of full thickness three-dimensional myocardial grafts by means of tissue engineering is limited by the impeded cellular viability in unperfused in vitro systems. We introduce a novel concept of pulsatile tissue culture perfusion to promote ubiquitous cellular viability and metabolism. METHODS: In a novel bioreactor we established pulsatile flow through the embedded three-dimensional tissue culture. Fibrin glue served as the ground matrix wherein neonatal rat cardiomyocytes were inoculated. Fluor-Deoxy-Glucose-Positron Emission-Tomography (FDG-PET) and life/dead assays were employed for comparative studies of glucose uptake resp. cell viability. RESULTS: A solid 8 mm thick structure resulted. Cellular viability significantly increased in the perfused chambers. We observed centripetal migration of the embedded cardiomyocytes to the site of the core vessel. However, cellular viability was high in the periphery of the tissue block too. FDG-PET revealed enhanced metabolic activity in perfused chambers. CONCLUSIONS: The present concept is highly effective in enhancing cellular viability and metabolism in a three-dimensional tissue culture environment. It could be utilized for various co-culture systems and the generation of viable tissue grafts. PMID- 14559015 TI - Preparation and modification of N-(2-hydroxyl) propyl-3-trimethyl ammonium chitosan chloride nanoparticle as a protein carrier. AB - N-(2-hydroxyl) propyl-3-trimethyl ammonium chitosan chloride (HTCC) is water soluble derivative of chitosan (CS), synthesized by the reaction between glycidyl trimethyl-ammonium chloride and CS. HTCC nanoparticles have been formed based on ionic gelation process of HTCC and sodium tripolyphosphate (TPP). Bovine serum albumin (BSA), as a model protein drug, was incorporated into the HTCC nanoparticles. HTCC nanoparticles were 110-180 nm in size, and their encapsulation efficiency was up to 90%. In vitro release studies showed a burst effect and a slow and continuous release followed. Encapsulation efficiency was obviously increased with increase of initial BSA concentration. Increasing TPP concentration from 0.5 to 0.7 mg/ml promoted encapsulation efficiency from 46.7% to 90%, and delayed release. As for modified HTCC nanoparticles, adding polyethylene glycol (PEG) or sodium alginate obviously decreased the burst effect of BSA from 42% to 18%. Encapsulation efficiency was significantly reduced from 47.6% to 2% with increase of PEG from 1.0 to 20.0 mg/ml. Encapsulation efficiency was increased from 14.5% to 25.4% with increase of alginate from 0.3 to 1.0 mg/ml. PMID- 14559016 TI - Chitin/PLGA blend microspheres as a biodegradable drug delivery system: a new delivery system for protein. AB - Novel chitin/PLGAs and chitin/PLA based microspheres were developed for the delivery of protein. These biodegradable microspheres were prepared by polymers blending and wet phase-inversion methods. The parameters such as selected non solvents, temperature of water and ratio of polylactide to polyglycolide were adjusted to improve thermodynamic compatibility of individual polymer (chitin and PLGAs or chitin/PLA), which affects the hydration and degradation properties of the blend microspheres. Triphasic pattern of drug release model is observed from the release of protein from the chitin/PLGAs and chitin/PLA microspheres: the initially fast release (the first phase), the following slow release (the second phase) and the second burst release (the third phase). Formulations of the blends, which are based on the balance among the hydration rate of the chitin phase and degradation of chitin/PLA and PLGA phase, can lead to a controllable release of bovine serum albumin (BSA). In conclusion, such a chitin/PLGA 50/50 microsphere is novel and interesting, and may be used as a protein delivery system. PMID- 14559017 TI - Preparation and characterization of poly(lactic acid)-poly(ethylene glycol) poly(lactic acid) (PLA-PEG-PLA) microspheres for controlled release of paclitaxel. AB - Microspheres of a new kind of copolymer, poly(lactic acid)-poly(ethylene glycol) poly(lactic acid) (PLA-PEG-PLA), are proposed in the present work for clinical administration of an antineoplastic drug paclitaxel with hypothesis that incorporation of a hydrophilic PEG segment within the hydrophobic PLA might facilitate the paclitaxel release. Paclitaxel-loaded PLA-PEG-PLA microspheres of various compositions were prepared by the solvent extraction/evaporation method. Characterization of the microspheres was then followed to examine the particle size and size distribution, the drug encapsulation efficiency, the colloidal stability, the surface chemistry, the surface and internal morphology, the drug physical state and its in vitro release behavior. The effects of polymer types, solvents and drug loading were investigated. It was found that in the microspheres the PEG segment was homogeneously distributed and caused porosity. Significantly faster release from PLA-PEG-PLA microspheres resulted in comparison with the PLGA counterpart. Incorporation of water-soluble solvent acetone in the organic solvent phase further increased the porosity of the PLA-PEG-PLA microspheres and facilitated the drug release. A total of 49.6% sustained release of paclitaxel within 1 month was achieved. Potentially, the presence of PEG on the surface of PLA-PEG-PLA microspheres could improve their biocompatibility. PLA PEG-PLA microspheres could thus be promising for the clinical administration of highly hydrophobic antineoplastic drugs such as paclitaxel. PMID- 14559018 TI - Mixing of acrylic bone cement: effect of oxygen on setting properties. AB - The present study investigates the effect of different mixing methods on the setting properties of bone cement. It was found that vacuum mixing decreased the setting time of the bone cement by nearly 2 min (10%), compared to mixing in air. Two additional experiments, in which the bone cement powders were purged with argon or oxygen, and mixed with the methyl methacrylate monomer, revealed that oxygen concentrations in the bone cement had a great effect on the setting time. The setting time increases significantly as the oxygen concentration increases, which suggests that the decrease in the setting time by vacuum mixing may be attributed to the lower oxygen levels present in the mixer. No significant effect was observed on dough time or maximum exothermic temperature by varying oxygen concentrations in the bone cement mixer. PMID- 14559019 TI - In vivo conjunctival reconstruction using modified PLGA grafts for decreased scar formation and contraction. AB - The in vivo reconstruction of conjunctiva was investigated by using modified poly(lactide-co-glycolide) (PLGA) 50/50 scaffolds. The porous PLGA matrices were prepared by a solvent-casting particulate-leaching method with NaCl, then modified with collagen, hyaluronic acid (HA) or/and human amniotic membrane (AM) component. The growth of corneal epithelial cells and human stromal fibroblasts on the scaffolds was investigated in vitro. All the modified PLGA scaffolds demonstrated enhanced cell adhesion and proliferation as compared to PLGA untreated, and the number of cells proliferated after 1 week was increased in the order of PLGA 0.84), triglycerides (P > 0.69), non esterified fatty acids (NEFA, P > 0.53), or glycerol (P > 0.33). Leptin at the intermediate and high doses depressed adipose expression of both PPARgamma1 (P < 0.06) and PPARgamma2 (P < 0.01). In a second study, we used a paired-feeding experimental design to determine the effects of a higher dose of leptin (0.05 mg. kg(-1). day(-1)) on serum metabolites and PPAR expression in selected tissues. At this dose, leptin increased (P < 0.0001) serum NEFA concentrations relative to both the ad libitum and pair-fed control groups. However, in this study, there was no difference in the expression of PPARgamma1 in adipose tissue, but PPARgamma2 mRNA was upregulated by leptin (P < 0.08). In contrast, leptin had no impact on the expression of PPARalpha in liver, skeletal muscle or adipose tissue. Adipose tissue explants were also incubated with leptin to assess the effect on PPARgamma expression, in vitro. The abundance of PPARgamma1 mRNA (P < 0.05) was increased after 24 hr of exposure, but the effect of leptin on gamma2 was not significant (P > 0.24). The lipolytic effect of leptin was also evaluated in vitro using isolated adipocytes. In keeping with the increase in serum NEFA concentrations in vivo, leptin stimulated lipolysis in vitro, increasing glycerol concentrations in the medium to about 219% of that in basal (non-treated) culture medium after 8 hr of incubation. Collectively, the data presented herein indicate that leptin modulates lipid metabolism in the pig, but that PPARalpha expression is not a parallel target of leptin as it is in rodent models. The regulation of PPARgamma by leptin seems complex in that it varied in relation to dose in vivo, and may be impacted by in vitro vs. in vivo circumstances. PMID- 14559109 TI - Dietary trans-18:1 raises plasma triglycerides and VLDL cholesterol when replacing either 16:0 or 18:0 in gerbils. AB - To compare the relative impact of trans-18:1 with the two main dietary saturated fatty acids it replaces, plasma lipid response was assessed in Mongolian gerbils fed diets rich in 16:0 (24%en),18:0 (10%en), or trans-18:1 (4 or 6%en). The diets were designed such that the 18:0-rich diet substituted 7%en as 18:0 for 16:0, whereas 4%en and 6%en from trans-18:1 was substituted for 16:0 in the two trans diets. The control group was fed a diet formulated according to the fatty acid balance of American Heart Association (AHA), but provided 40%en as fat. Gerbils (n = 10 per dietary group) were fed one of the five diets for 8 weeks. The control diet, with 4 times the polyunsaturated fatty acids (PUFA) content and a P:S ratio about 10 times greater than the test diets, resulted in the lowest plasma TC, LDL cholesterol (LDL-C) and VLDL cholesterol (VLDL-C). Among the test diets, plasma TC and TG were lowest with the 18:0-rich diet. TC in gerbils fed the 16:0-rich diet and 4%en-trans were 20% higher than the 18:0-rich diet, while the 6%en-trans diet was 35% higher. VLDL-C was significantly higher in the 6%en trans diet compared to all other groups at 8 weeks. Both trans fatty acid diets elevated plasma TG approximately 2- and 3-fold, respectively, compared to the 16:0-rich and 18:0-rich diets at 8 weeks. Further, plasma TG continued to rise over time with trans fatty acids compared to 16:0 or 18:0. Thus, in the fatty acid-sensitive gerbil, impaired TG metabolism represents a major aspect of the hyperlipemia caused by trans fatty acid substitution for major saturated fatty acids. PMID- 14559110 TI - S-adenosylmethionine (SAMe) protects against acute alcohol induced hepatotoxicity in mice. AB - Although S-Adenosylmethionine (SAMe) has beneficial effects in many hepatic disorders, the effects of SAMe on acute alcohol-induced liver injury are unknown. In the present study, we investigated effects of SAMe on liver injury in mice induced by acute alcohol administration. Male C57BL/6 mice received ethanol (5 g/kg BW) by gavage every 12 hrs for a total of 3 doses. SAMe (5 mg/kg BW) was administrated i.p. once a day for three days before ethanol administration. Subsequent serum ALT level, hepatic lipid peroxidation, enzymatic activity of CYP2E1 and hepatic mitochondrial glutathione levels were measured colorimetrically. Intracellular SAMe concentration was measured by high performance liquid chromatography (HPLC). Histopathological changes were assessed by H&E staining. Our results showed that acute ethanol administration caused prominent microvesicular steatosis with mild necrosis and an elevation of serum ALT activity. SAMe treatment significantly attenuated the liver injury. In association with the hepatocyte injury, acute alcohol administration induced significant decreases in both hepatic SAMe and mitochondrial GSH levels along with enhanced lipid peroxidation. SAMe treatment attenuated hepatic SAMe and mitochondrial GSH depletion and lipid peroxidation following acute alcohol exposure. These results demonstrate that SAMe protects against the liver injury and attenuates the mitochondrial GSH depletion caused by acute alcohol administration. SAMe may prove to be an effective therapeutic agent in many toxin induced liver injuries including those induced by alcohol. PMID- 14559111 TI - Inulin stimulates NO synthesis via activation of PKC-alpha and protein tyrosine kinase, resulting in the activation of NF-kappaB by IFN-gamma-primed RAW 264.7 cells. AB - Inulin, an active component of Chicorium intybus root, has been shown to stimulate the growth of bifidobacteria, and inhibit colon carcinogenesis. NO mediates a number of the host-defense functions of activated macrophages, including antimicrobial and tumoricidal activity. We examined the effect of inulin on the synthesis of NO in RAW 264.7 cells. Inulin alone had no effect, whereas inulin with IFN-gamma synergistically increased the NO production and inducible NO synthase (iNOS) expression in RAW 264.7 cells. Synergy between IFN gamma and inulin was mainly dependent on inulin-induced TNF-alpha secretion. Also, protein kinase C (PKC)-alpha was involved in the inulin-induced NO production. Inulin-mediated NO production was inhibited by the protein tyrosine kinase (PTK) inhibitor, tyrphostin AG126. Since iNOS gene transcriptions have been shown to be under the control of the NF-kappaB/Rel family of transcription factors, we assessed the effect of inulin on NF-kappaB/Rel using an EMSA. Inulin produced strong induction of NF-kappaB/Rel binding, whereas AP-1 binding was slightly induced in RAW 264.7 cells. Inulin stimulated phosphorylation and degradation of IkappaB-alpha. These results suggest that in IFN-gamma-primed RAW 264.7 cells inulin might stimulate NO synthesis via activation of PKC-alpha and PTK, resulting in the activation of NF-kappaB. PMID- 14559112 TI - Protective effect of vitamin C against the ethanol mediated toxic effects on human brain glial cells. AB - It is now known that chronic consumption of excessive amounts of alcohol is a major source of social and medical problems. Ethanol-mediated glial cell activation may lead to neuron damage in many ways, including the formation of free radicals and production of pro-inflammatory molecules. Vitamin C (vit-C) is an essential dietary nutrient required as a co-factor for many enzymes and a very efficient antioxidant, protecting cells against free radical-mediated damage. The objective of this study was to evaluate the protective effects of vit-C on glial cell activation and viability against ethanol-mediated toxicity. Human brain astrocyte cells (HA) were exposed to ethanol (0, 50, and 350 mmol/L) for 24 h. We found that glial cells incubated with different concentrations of vit-C increase their vit-C in a dose-dependent manner. HA incubated with 0, 50 or 350 mmol/L of ethanol for up to 24 h showed toxic effects that were proportional to the levels of ethanol in the medium, HA showed increased levels of heat shock protein (Hsp70). However, cells enriched with vit-C before being exposed to ethanol, were better protected against the alcohol-mediated toxicity than non-supplemented cells, and showed significantly lower concentrations of Hsp70. Ethanol also caused increased expression of cyclooxygenase-2 (COX-2) and synthesis of prostaglandin E2 (PGE2), which were reduced by vit-C. In summary, HA supplemented with vit-C were significantly more resistant to the ethanol-mediated toxic effects. PMID- 14559113 TI - Lipopolysaccharides of Vibrio cholerae. I. Physical and chemical characterization. AB - Vibrio cholerae is the causative organism of the disease cholera. The lipopolysaccharide (LPS) of V. cholerae plays an important role in eliciting the antibacterial immune response of the host and in classifying the vibrios into some 200 or more serogroups. This review presents an account of our up-to-date knowledge of the physical and chemical characteristics of the three constituents, lipid-A, core-polysaccharide (core-PS) and O-antigen polysaccharide (O-PS), of the LPS of V. cholerae of different serogroups including the disease-causing ones, O1 and O139. The structure and occurrence of the capsular polysaccharide (CPS) on V. cholerae O139 have been discussed as a relevant topic. Similarity and dissimilarity between the structures of LPS of different serogroups, and particularly between O22 and O139, have been analysed with a view to learning their role in the causation of the epidemic form of the disease by avoiding the host defence mechanism and in the evolution of the newer pathogenic strains in future. An idea of the emerging trends of research involving the use of immunogens prepared from synthetic oligosaccharides that mimic terminal epitopes of the O-PS of V. cholerae O1 in the development of a conjugate anti cholera vaccine is also discussed. PMID- 14559114 TI - Tryptophan hydroxylase expression in human skin cells. AB - We attempted to further characterize cutaneous serotoninergic and melatoninergic pathways evaluating the key biosynthetic enzyme tryptophan hydroxylase (TPH). There was wide expression of TPH mRNA in whole human skin, cultured melanocytes and melanoma cells, dermal fibroblasts, squamous cell carcinoma cells and keratinocytes. Gene expression was associated with detection of TPH immunoreactive species by Western blotting. Characterization of the TPH immunoreactive species performed with two different antibodies showed expression of the expected protein (55-60 kDa), and of forms with higher and lower molecular weights. This pattern of broad spectrum of TPH expression including presumed degradation products suggests rapid turnover of the enzyme, as previously reported in mastocytoma cells. RP-HPLC of skin extracts showed fluorescent species with the retention time of serotonin and N-acetylserotonin. Immunocytochemistry performed in skin biopsies localized TPH immunoreactivity to normal and malignant melanocytes. We conclude that while the TPH mRNA and protein are widely expressed in cultured normal and pathological epidermal and dermal skin cells, in vivo TPH expression is predominantly restricted to cells of melanocytic origin. PMID- 14559115 TI - Congenital afibrinogenemia: intracellular retention of fibrinogen due to a novel W437G mutation in the fibrinogen Bbeta-chain gene. AB - Congenital afibrinogenemia is a rare autosomal recessive coagulation disorder characterised by hemorrhagic manifestations of variable entity and by severe plasma fibrinogen deficiency. Among the 31 afibrinogenemia-causing mutations so far reported, only 2 are missense mutations and both are located in the fibrinogen Bbeta-chain gene. Direct sequencing of the fibrinogen gene cluster in two afibrinogenemic Iranian siblings revealed a novel homozygous T>G transversion in exon 8 (nucleotide position 8025) of the fibrinogen Bbeta-chain gene. The resulting W437G missense mutation involves a highly conserved amino acid residue, located in the C-terminal globular D domain. The role of the W437G amino acid substitution on fibrinogen synthesis, folding, and secretion was assessed by in vitro expression experiments in COS-1 cells, followed by qualitative and quantitative analyses of intracellular and secreted mutant fibrinogen. Results of both pulse-chase experiments and enzyme-linked immunosorbent assays demonstrated intracellular retention of the mutant W437G fibrinogen and marked reduction of its secretion. These data, besides elucidating the pathogenetic role of the W437G mutation in afibrinogenemia, underline the importance of the Bbeta-chain D domain in fibrinogen folding and secretion. PMID- 14559116 TI - The alpha-L-iduronidase mutations R89Q and R89W result in an attenuated mucopolysaccharidosis type I clinical presentation. AB - Mucopolysaccharidosis type I (MPS I; McKusick 25280; Hurler syndrome, Hurler Scheie syndrome and Scheie syndrome) is caused by a deficiency in the lysosomal hydrolase, alpha-L-iduronidase (EC 3.2.1.76). MPS I patients present within a clinical spectrum bounded by the extremes of Hurler and Scheie syndromes. The alpha-L-iduronidase missense mutations R89Q and R89W were investigated and altered an important arginine residue proposed to be a nucleophile activator in the catalytic mechanism of alpha-L-iduronidase. The R89Q alpha-L-iduronidase mutation was shown to result in a reduced level of alpha-L-iduronidase protein (< or =10% of normal control) compared to a normal control level of alpha-L iduronidase protein that was detected for the R89W alpha-L-iduronidase mutation. When taking into account alpha-L-iduronidase specific activity, the R89W mutation had a greater effect on alpha-L-iduronidase activity than the R89Q mutation. However, overall the R89W mutation produced more residual alpha-L-iduronidase activity than the R89Q mutation. This was consistent with MPS I patients, with an R89W allele, having a less severe clinical presentation compared to MPS I patients with either a double or single allelic R89Q mutation. The effects of the R89Q and R89W mutations on enzyme activity supported the proposed role of R89 as a nucleophile activator in the catalytic mechanism of alpha-L-iduronidase. PMID- 14559117 TI - The RING-H2 protein RNF11 is differentially expressed in breast tumours and interacts with HECT-type E3 ligases. AB - A breast cancer-associated mRNA originally cloned as a 475-bp partial cDNA from a library enriched for tumour cDNAs [Oncogene 16 (1998) 327] is expressed at high levels in breast and prostate cancer cells. Immunohistochemical analysis indicates that the protein is expressed in primary breast tumours. We used RT-PCR to generate a full-length 2852 nt mRNA sequence that includes the hypothetical open reading frame (ORF) for human RNF11. Our analysis shows that RNF11 encodes modular domains and motifs likely to interact with other proteins involved in oncogenesis. Chief among these are the RING-H2 finger domain that could facilitate the degradation of specific substrate(s) involved in oncogenesis and the PY motif which binds to WW-domain proteins, several of which are known to be E3 ubiquitin ligases. Our GST-pulldown and immunoprecipitation results indicate that RNF11 interacts with the E3 ligase AIP4 when coexpressed with RNF11 in mammalian cells. PMID- 14559118 TI - Diabetes induces metabolic adaptations in rat liver mitochondria: role of coenzyme Q and cardiolipin contents. AB - Several studies have been carried out to evaluate the alterations in mitochondrial functions of diabetic rats. However, results are sometimes controversial, since experimental conditions diverge, including age and strain of used animals. The purpose of this study was to evaluate the metabolic modifications in liver mitochondria, both in the presence of severe (STZ-treated rats) and mild hyperglycaemia [Goto-Kakizaki (GK) rats], when compared with control animals of similar age. Moreover, metabolic alterations were evaluated also at initial and advanced stages of the disease. We observed that both models of diabetes (type 1 and type 2) presented a decreased susceptibility of liver mitochondria to the induction of permeability transition (MPT). Apparently, there is a positive correlation between the severity of diabetes mellitus (and duration of the disease) and the decline in the susceptibility to MPT induction. We also found that liver mitochondria isolated from diabetic rats presented some metabolic adaptations, such as an increase in coenzyme Q and cardiolipin contents, that can be responsible for the observed decrease in the susceptibility to multiprotein pore (MPTP) opening. PMID- 14559119 TI - Increased formation of methylglyoxal and protein glycation, oxidation and nitrosation in triosephosphate isomerase deficiency. AB - Triosephosphate isomerase deficiency is associated with the accumulation of dihydroxyacetonephosphate (DHAP) to abnormally high levels, congenital haemolytic anaemia and a clinical syndrome of progressive neuromuscular degeneration leading to infant mortality. DHAP degrades spontaneously to methylglyoxal (MG)--a potent precursor of advanced glycation endproducts (AGEs). MG is detoxified to D-lactate intracellularly by the glyoxalase system. We investigated the changes in MG metabolism and markers of protein glycation, oxidation and nitrosation in a Hungarian family with two germline identical brothers, compound heterozygotes for triosephosphate isomerase deficiency, one with clinical manifestations of chronic neurodegeneration and the other neurologically intact. The concentration of MG and activity of glyoxalase I in red blood cells (RBCs) were increased, and the concentrations of D-lactate in blood plasma and D-lactate urinary excretion were also increased markedly in the propositus. There were concomitant increases in MG derived AGEs and the oxidative marker dityrosine in hemoglobin. Smaller and nonsignificant increases were found in the neurologically unaffected brother and parents. There was a marked increase (15-fold) in urinary excretion of the nitrosative stress marker 3-nitrotyrosine in the propositus. The increased derangement of MG metabolism and associated glycation, oxidative and nitrosative stress in the propositus may be linked to neurodegenerative process in triosephosphate isomerase deficiency. PMID- 14559120 TI - Inhibition by Aplidine of the aggregation of the prion peptide PrP 106-126 into beta-sheet fibrils. AB - Aplidine, a cyclic peptide, from the tunicate Aplidium albican, prevents the in vitro aggregation into beta-sheet containing fibrils of the prion peptide 106-126 when co-incubated in a 1:1 molar ratio. The blocking of fibril formation induced by Aplidine has clear sequence specificity, being much stronger for the 106-126 prion peptide than for the beta-amyloid 25-35 peptide. In addition to the known ability of Aplidine to cross the plasmatic membrane, these results indicate that Aplidine is a potential leading compound for the development of therapeutic blockers of prion aggregation. PMID- 14559121 TI - Clinical perspectives and therapeutics of thrombolysis. AB - Reperfusion therapy with thrombolytic agents has been a significant advancement in the management of patients with acute ST elevation myocardial infarction. The outcome of acute myocardial infarction has significantly improved by early application of thrombolytic therapy. Intracoronary streptokinase has been used for >30 years, but reawakening interest occurred in the early 1980s in the use of thrombolytic therapy to establish rapid reperfusion during an acute myocardial infarction. Initial studies aimed at direct intracoronary thrombolysis, but owing to its cumbersome process and requirement of an active round the clock cardiac catheterization laboratory, it has been replaced by regimens of intravenous thrombolytic therapy which is as efficacious as intracoronary administration. Consideration of thrombolytic therapy has become a standard treatment for patients presenting with acute ST elevation myocardial infarction and various well-controlled trials have demonstrated the importance of both early and full reperfusion in improving clinical outcome in the setting of acute myocardial infarction. The subject of intravenous thrombolysis is perhaps the most rapidly evolving area in the management of acute myocardial infarction patients in the past decade. The current review focuses on the thrombolysis in the treatment of myocardial infarction and other conditions. PMID- 14559122 TI - Absence of clinical signs of cardiac denervation after percutaneous myocardial laser revascularization. AB - BACKGROUND: Percutaneous myocardial revascularization with laser (PMR) is a catheter-based technique that has generated much interest in the treatment of patients with severe coronary artery disease. Several mechanisms have been proposed to explain the reported clinical benefits of this technique. Cardiac autonomic denervation is among these. METHODS: We studied 32 consecutive patients with chronic severe angina not suitable for other revascularization approaches. Canadian Class Society (CCS) class clinical assessment, Naughton exercise stress test, and scintigraphic myocardial perfusion were evaluated before and 1 and 6 months after PMR. Ewing's autonomic tests, heart rate variability (HRV), and plasmatic catecholamines were assessed before revascularization in non-diabetic and diabetic patients and repeated 1 month after PMR in 13 non-diabetics. A psychological test was carried out before PMR to evaluate the attitude towards this new procedure. RESULTS: All the markers of autonomic cardiac control were unmodified after the procedure. Moreover, scintigraphic perfusion measured in the lasered areas was similar before and after the procedure. On the other hand, the clinical conditions significantly improved (CCS class from 3+/-0.8 to 1.9+/-0.9 at 1 month, P<0.01) and the exercise-related ischemic threshold was significantly better (from 311+/-28 to 453+/-51 s, P<0.05). The patients with a psychologically 'positive' expectation for this new procedure had results comparable to those with a 'negative' expectation. CONCLUSIONS: PMR improves symptoms in patients with end-stage coronary artery disease in the absence of any detectable clinical sign of heart denervation. PMID- 14559123 TI - Serum myoglobin/carbonic anhydrase III ratio as a marker of reperfusion after myocardial infarction. AB - BACKGROUND: Coronary patency is important for short- and long-term outcome after myocardial infarction. Serum myoglobin concentration is a sensitive marker of myocardial damage and its specificity can be improved by simultaneous measurement of carbonic anhydrase III, a skeletal muscle marker. In the present study we evaluated the role of myoglobin/carbonic anhydrase III ratio as a non-invasive marker of reperfusion. METHODS: We measured myoglobin, carbonic anhydrase III and creatine kinase MB-fraction release serially in 29 patients with acute myocardial infarction treated with thrombolysis and in 28 patients treated with primary coronary angioplasty. RESULTS: Thrombolytic therapy was followed by a 9.1+/-2.2 fold increase in myoglobin and 10.8+/-3.3-fold increase in creatine kinase MB fraction during the first hour of treatment, while carbonic anhydrase III remained unchanged. The peak value of myoglobin/carbonic anhydrase III ratio was found at 2 h and that of creatine kinase MB-fraction at 8 h after thrombolysis. Knowledge of the reperfusion time point during primary angioplasty and follow-up of cardiac markers revealed that cut-off points of 3 and 10 h for the peak values of myoglobin/carbonic anhydrase III ratio and creatine kinase MB-fraction can be used as indicators for reperfusion, respectively. Myoglobin/carbonic anhydrase III ratio measured before treatment and at 2 and 4 h after the onset of treatment screened 23 of those 25 patients with probable reperfusion after thrombolysis. CONCLUSIONS: We conclude that measuring myoglobin/carbonic anhydrase III ratio during the first hours after initiation of thrombolysis may be useful in evaluating the success of reperfusion after acute myocardial infarction. PMID- 14559124 TI - Eighteen-year experience with echinococcosus of the heart: clinical and echocardiographic features in 14 patients. AB - We present a retrospective analysis of clinical symptoms and echocardiographic features in 14 patients having cardiac hydatic cysts and screened during the 18 last years, with surgical correlations in 13. The heart was the sole location of the cyst in six patients. The revealing symptoms were precordial pain (six patients), dyspnea (two patients). A systolic murmur of false pulmonic stenosis was present in three patients. In 13 patients, two-dimensional echocardiographic images of hydatic cysts were those of masses of liquid content with a well contrasted capsule, which was single in eight patients and multiple in five, one of which had a honeycomb appearance. The mass was solid and calcified in one patient. The outlines of the cysts were particularly sharp on transesophageal echocardiograms (six patients). The cyst size ranged from 1.8 to 11 cm in diameter. Cysts were located in the intramyocardium in 10 patients (left ventricle in four patients, interventricular septum in four patients, right ventricle in two patients), the pericardium in three and the mediatinum in one. There were a Doppler pulmonary valve gradient in three patients. Computed tomography (eight patients) and coronary arteriography (seven patients) have no additional diagnosis value than echocardiography. Nuclear magnetic resonance imaging (three patients) was helpful in determining extracardiac extension of the cysts. Transthoracic and transesophageal two-dimensional echocardiography have a central role in diagnosing cardiac hydatic cysts. PMID- 14559125 TI - sICAM-1 correlates with myocardial ICAM-1 expression in dilated cardiomyopathy. AB - BACKGROUND: Dilated cardiomyopathy (DCM) is etiopathogenically linked to intramyocardial inflammation, which is reflected by ICAM-1 abundance. We investigated whether soluble ICAM-1 (sICAM-1) levels in the sera of DCM patients are associated with intramyocardial ICAM-1 expression. METHODS: Immunohistochemically detected ICAM-1 expression was quantified semiquantitatively in endomyocardial biopsies from DCM patients (n=45; n=17 females; age: 48+/-15 years) and from n=12 donor hearts (controls) by a human observer (baseline vs. enhanced expression) and quantitatively by a digital image analysis (DIA) system. The DIA-measured qualities were area fraction (AF), surface-volume ratio (SVR) and integral optical density (ID). The sICAM-1 levels of the DCM patients and n=12 healthy volunteers (controls) were measured by ELISA (means of duplicate measurements). Intramyocardial ICAM-1 expression and sICAM-1 levels were compared in these DCM patients. RESULTS: Of the DCM patients, n=24 (53%) demonstrated statistically higher sICAM-1 levels compared to controls (>198 ng/ml). By semiquantitative and quantitative DIA evaluation, endothelial ICAM-1 abundance was present in n=25 (56%) of the DCM biopsies. sICAM-1 correlated significantly (P<0.001) both with the semiquantitatively assessed and the DIA measured ICAM-1-AF, the ICAM-1-SVR and the ICAM-1-ID. The positive predictive value of sICAM-1 measurements for intramyocardial ICAM-1 abundance was 96%, and the negative predictive value was 71%, with a receiver operating characteristic area under the curve of 0.93. Furthermore, sICAM-1 levels correlated with intramyocardial T-lymphocytic (CD2+/CD3+) infiltrates (P<0.03). CONCLUSIONS: Measurement of non-invasively obtained sICAM-1 reliably reflects intramyocardial ICAM-1 expression and may therefore serve as a non-invasive marker of inflammatory activity in DCM. PMID- 14559126 TI - National economic impact of tirofiban for unstable angina and myocardial infarction without ST elevation; example from the United Kingdom. AB - BACKGROUND: Acute coronary syndromes without ST elevation are a major health and economic burden. Treatments such as glycoprotein IIb/IIIa antagonists like tirofiban reduce the risk of complications but the cost impact of these agents including cost offsets of avoiding complications are needed particularly in Europe. METHODS: We used treatment patterns from the Prospective Registry of Acute Ischemic Syndromes in the UK, risk reductions derived from the PRISM-PLUS trial and cost estimates from the CHKS database to estimate the impact of tirofiban on PRAIS-UK patients with and without complications and subgroups at higher risk of complications. These subgroups (and proportions) were patients: (1) aged 60 or over with abnormal electrocardiograms (58%), (2) with ST depression or bundle branch block on admission (30%) and (3) with ST depression, bundle branch block or MI on admission (37%). RESULTS: Total cost of care in the UK at 6 months for the estimated 87339 acute coronary syndromes admissions annually was pound 213 million, which would increase by pound 33 million (15.7%) if tirofiban were given to all patients, avoiding 2422 complications at a mean cost per event avoided of pound 13388. Among the subgroups, the mean cost per event avoided ranges from pound 10856 for subgroup 1 to pound 5953 for subgroup 3. Treating the latter subgroup, would avoid 1977 events at a cost of pound 12 million (5.5%). CONCLUSION: The use of tirofiban in the UK to treat acute coronary syndromes patients without ST elevation provides an important therapeutic advantage at modest proportional increase in cost, particularly if targeted to higher risk subgroups as recommended in the European guidelines. PMID- 14559127 TI - Treatment of acute inflammatory cardiomyopathy with intravenous immunoglobulin ameliorates left ventricular function associated with suppression of inflammatory cytokines and decreased oxidative stress. AB - Although an autoimmune mechanism has been postulated for myocarditis and dilated cardiomyopathy, immunosuppressive agents had not been shown to be effective. Potential benefits of intravenous immunoglobulin (IVIg) in the therapy of patients with myocarditis and recent onset of dilated cardiomyopathy were reported. Also, experimental studies showed that IVIg is an effective therapy for viral myocarditis by antiviral and anti-inflammatory effects. Accordingly, in the current study, the effects of IVIg in the patients were investigated with the analyses of inflammatory cytokines and oxidative stress. Nine patients (six in myocarditis, three in acute dilated cardiomyopathy) were treated with high-dose intravenous IVIg (1-2 g/kg, over 2 days). All were hospitalized with New York Heart Association (NYHA) class III to IV heart failure, left ventricular ejection fraction (LVEF) <40%, and symptoms for <6 months at the time of presentation. Five patients were diagnosed using endomyocardial biopsy. LVEF determined by echocardiography improved from 19.0+/-7.5% (mean+/-S.D.) at baseline to 35.4+/ 9.1% at follow up (12.2+/-5.8 days after the treatment) (P<0.01). C-reactive protein and plasma inflammatory cytokines (tumor necrosis factor-alpha and interleukin-6) were decreased by this treatment. In addition, plasma level of thioredoxin, which regulates the cellular state of oxidative stress, was decreased by the treatment. All nine patients improved functionally to NYHA class I to II, and were discharged without side-effects. There have been no subsequent hospitalizations for heart failure during the course of follow-up (3 months-4.5 years). LVEF improved 16% of EF in the patients with myocarditis and acute dilated cardiomyopathy with the reduction of cytokines associated with improvement of oxidative stress state by high-dose of IVIg. Thus, IVIg seems to be a promising agent in the therapy of acute inflammatory cardiomyopathy in view of not only suppression of inflammatory cytokines but a reduction of oxidative stress. PMID- 14559128 TI - Fibrillatory wave amplitude as a marker of left atrial and left atrial appendage function, and a predictor of thromboembolic risk in patients with rheumatic mitral stenosis. AB - BACKGROUND: Patients with mitral stenosis, especially those with atrial fibrillation, are at increased risk for thromboembolic complications. Size of the left atrium, left atrial appendage dysfunction and severity of mitral stenosis are known risk factors for thromboembolism in patients with mitral stenosis. It has been postulated that F-wave amplitude on surface ECG is correlated with left atrial size, left atrial appendage function, and risk of thromboembolism in patients with nonrheumatic atrial fibrillation. The aims of this study were as follows: (1) to examine the relationship between surface ECG F-wave amplitude and left atrial appendage function, and (2) to assess the clinical significance of F wave amplitude as it relates to risk of thromboembolism in a group of patients with rheumatic mitral stenosis. MATERIAL AND METHODS: One hundred patients with rheumatic mitral stenosis and permanent atrial fibrillation but without moderate to severe mitral insufficiency were investigated by transthoracic and transesophageal echocardiography. Two groups were formed according to the presence of a coarse (Group 1; n=60; F-wave in lead V1> or =1 mm) or fine (Group 2; n=40; F-wave in lead V1<1 mm) F-wave on surface ECG. RESULTS: Comparison showed that Group 1 had significantly smaller mitral valve area (1.44+/-0.6 versus 1.7+/-0.74 cm2, respectively; P<0.05), lower peak left atrial appendage flow velocity (18.8+/-2.1 versus 25.6+/-1.9 cm/s, respectively; P<0.005), higher grade spontaneous echo contrast (2.05+/-1.44 versus 0.98+/-1.14, respectively; P<0.0001), and higher prevalence of thromboembolism (35% versus 12.5%, respectively; P<0.012). Multiple logistic regression analysis revealed that mitral valve area, left atrial appendage peak velocity, and coarse F-wave were independent clinical risk factors for thromboembolism in this patient group. CONCLUSION: The data suggest that presence of a coarse F-wave on surface ECG is associated with left atrial appendage dysfunction, and indicates higher thromboembolic risk in patients with predominant rheumatic mitral stenosis. PMID- 14559129 TI - Myocardial perfusion in real-time using power modulation. In vivo evidence for microcirculatory damage after acute myocardial infarction. AB - BACKGROUND AND OBJECTIVES: In addition to the myocardium, the microvasculature may be also damaged in acute myocardial infarction. The aim was to evaluate the capability of myocardial contrast echocardiography in the detection of microvasculature damage after myocardial infarction. PATIENTS AND METHODS: Twelve patients with recent acute myocardial infarction and five control subjects with normal coronary arteries and without history of myocardial infarction were studied. Myocardial contrast echocardiography with power modulation was performed, and quantitative data were measured off-line. Power modulation uses a combination of low (0.1) and high (1.7) mechanical indexes, allowing a real-time evaluation of myocardial perfusion. Contrast agent was administered as a 3-min bolus. The quantitative analysis was performed off-line by a different blinded investigator. The refilling velocity was calculated as the difference between the peak myocardial refilling value and the value at 1 s after the impulse divided by the time from the first second after the impulse to the peak refilling value. RESULTS: Eighty-one myocardial segments (75%) were analysed qualitatively and quantitatively in AMI patients, and 18 (60%) in control subjects (P=NS). The peak refilling intensity was not significantly different in patients and control subjects (6.62+/-5.85 vs. 7.53+/-4.06 dB, respectively). However, time to peak refilling intensity was significantly longer (5.25+/-1.57 vs. 4.00+/-0.53, P=0.004) and the velocity of refilling was significantly lower (2.74+/-5.34 vs. 6.58+/-8.02, P=0.028) in patients with myocardial infarction. CONCLUSION: There is microvasculature damage after myocardial infarction that is reflected as a delayed velocity of refilling in myocardial contrast echocardiography. PMID- 14559130 TI - The pre-existence of an acute coronary event predicts differences in biological parameters and clinical evolution among patients with longstanding stable angina. AB - BACKGROUND: To date, patients displaying stable angina with or without a previous acute coronary event have been grouped together and are offered nowadays, similar clinical approach. However, from clinical experience, increasing doubt exists whether they behave clinically in the same way. In the current study, we investigated whether differences might exist in the clinical, angiographic and biological parameters between these two distinct clinical presentations. METHODS: The cardiovascular risk factor profile, clinical evolution, angiographic data, white blood cell (WBC) count and plasma concentrations of lipids and fibrinogen were compared between two groups of patients with longstanding stable angina. The first group consisted of 160 subjects who had undergone myocardial infarction (MI) as the first manifestation of coronary artery disease (CAD) and had no further coronary event. The second group was composed of 226 subjects who had effort-induced angina but never experienced an acute coronary event. The mean period between the first CAD manifestation and entry into the study was 7+/-6 years for MI group and 7+/-4 years for patients with uneventful angina. RESULTS: MI patients displayed 8% higher WBC count (P=0.038), 20% higher fibrinogen (P=0.001), 12% lower HDL cholesterol (P=0.02), 8% lower apoA1 (P=0.005) and 46% higher lipoprotein (a) (P=0.005) than patients who presented an uneventful clinical course. Upon multivariate logistic regression analysis, fibrinogen (OR=1.14; 95% CI=1.08-1.28; P=0.033), lipoprotein (a) (OR=1.19; 95% CI=1.05-1.34; P=0.0036) and HDL cholesterol levels (OR=0.97; 95% CI=0.94-0.99; P=0.0095) remained independently associated with the pre-existence of MI. Patients with prior MI evolved more frequently towards clinically refractory angina (P=0.04). In addition, fibrinogen levels were independently associated with evolution to clinically refractory angina (OR=1.03; 95% CI=1.005-1.550; P=0.0448). CONCLUSIONS: Stable CAD patients with a prior MI differ from those who display an uneventful effort-induced angina in their biological profile and their evolution towards clinically refractory angina. PMID- 14559131 TI - Pharmacokinetics and hemodynamic effects of the phosphodiesterase III inhibitor saterinone in patients with chronic heart failure. AB - The introduction of PDE III-inhibitors has been a major innovation for the intravenous drug treatment of patients suffering from heart failure. In this study, the pharmacokinetics and hemodynamic effects of the PDE III-inhibitor saterinone were examined in twelve male patients with severe chronic heart failure. METHODS: Saterinone was given by intravenous infusion for 24 hours at a rate of 1.5 microg/kg per min. According to a standardized protocol, blood samples were drawn for measurement of saterinone plasma levels and hemodynamic measurements were taken in order to analyze the correlation between plasma levels and hemodynamic effects. RESULTS: The correlation coefficient between the increase in cardiac index and the logarithms of saterinone plasma concentrations was r=0.827 (p=0.003). Between the logarithms of plasma concentrations and the decrease in pulmonary capillary wedge pressure a correlation coefficient of r=0.964 (p<0.001) was calculated for the first twelve hours of saterinone infusion. The decrease of saterinone plasma concentrations can be fitted to a three-compartment-model (half-lifes were 4.24 minutes for the alpha-phase, three hours for the beta-phase and 15.7 hours for the terminal phase). Saterinone induced maximal increases of 56.6% in cardiac index, 48.9% in stroke volume index, 28.4% in heart rate and maximal decreases of 17.3% in mean systemic blood pressure, 38.4% in mean pulmonary artery pressure, 74.2% in right atrial pressure, 46.9% in pulmonary capillary wedge pressure, 39.9% in systemic vascular resistance and 71.8% in pulmonary vascular resistance. CONCLUSION: Saterinone was demonstrated to be a safe, potent drug during an intravenous infusion over 24 hours at a rate of 1.5 microg/kg per min. PMID- 14559132 TI - Relationship between myocardial viability and the predischarge electrocardiographic pattern in patients with first anterior wall acute myocardial infarction. AB - BACKGROUND: The assessment of residual viability in the infarcted area after an acute myocardial infarction is relevant to subsequent management and prognosis. OBJECTIVE: The aim of this study was to investigate the correlation between myocardial viability after an acute anterior myocardial infarction (AMI) as assessed by low dose dobutamine stress echocardiography (LDDSE) and the electrocardiographic patterns of ST segment and T wave abnormalities at the end of the first week of the acute event. METHODS: Sixty-nine consecutive patients (51 men, 18 women, mean age+/-standard deviation=57+/-11 years) who admitted to our clinic due to a first episode of transmural AMI were included in this study. Two-dimensional echocardiography was performed to all patients during rest and low dose dobutamine administration at the end of the first week of admission (7+/ 2 days). Patients were classified into four groups according to ST segment and T wave morphology: group A, ST elevation < or =0.1 mV and negative T waves; group B, ST elevation < or =0.1 mV and positive T waves; group C, ST elevation > or =0.1 mV and negative T waves and group D, ST elevation > or =0.1 mV and positive T waves. RESULTS: Myocardial viability was detected more often in patients with isoelectric ST segments (22/24, 92%) than those with elevated ST segments (21/45, 47%) (P<0.001). Similarly patients with negative T waves had myocardial viability more frequently compared to those with positive T waves (32/45, 71% vs. 11/24, 46%, P<0.01). Seventeen (94%) of 18 patients in group A and 5 (83%) of six patients in group B had viable myocardium (P>0.05). Myocardial viability was found in 15 (56%) of 27 patients in group C and six (33%) of 18 patients in group D (P<0.01). As a marker of viable myocardium, isoelectricity of ST segment was specific (92%) but only moderately sensitive (51%), with a 92% positive predictive accuracy and a poor (53%) negative predictive value. T wave negativity was less specific but more sensitive than isoelectricity of ST segment for myocardial viability. CONCLUSION: The presence of isoelectric ST segment and negative T wave indicates a high probability of myocardial viability. However, absence of these electrocardiographic patterns does not exclude the presence of viable myocardium. PMID- 14559133 TI - Cardiac pacemaker lead extraction using conventional techniques:a single centre experience. AB - BACKGROUND: New methods of lead extraction using laser sheath devices are under evaluation but these techniques are not available in the majority of centres and have cost implications. Furthermore, in the absence of comparative randomised trials, registry experience with new devices must be judged against contemporary data using conventional methods. We report a single centre series of pacemaker lead extraction using conventional methods. METHODS: STUDY POPULATION: Attempted extraction of 165 leads during 95 procedures in 80 patients. Leads had been in place (dwell time) for a mean of 76 months (range 0.2-248.4 months). Indications for lead extraction: infection (41.1%), skin erosion (9.5%), advisory leads (12.6%), faulty leads (12.6%), other (24.2%). Extraction techniques: traction and/or locking stylets and dilator sheaths (89.7%), Byrd workstation (6.1%) and open thoracotomy (4.8%). RESULTS: Complete removal was achieved for 143 leads (86.7%), partial removal in 12 leads (7.3%) and 10 (6.1%) could not be removed. A shorter lead dwell time was associated with extraction success in both univariate (p=0.0004) and multivariate analyses (p<0.0001). There was a trend for a higher rate of success in atrial rather than ventricular leads (93.2% v 80.9%, p=0.052). Active fixation, patient gender, age and indication for lead extraction had no bearing on outcome. COMPLICATIONS: There were no deaths. Major complications occurred in 3 patients (3.2%): pericardial tamponade (1), pulmonary embolus (1) and stroke (1). Significant bleeding (requiring blood transfusion) occurred in 11 procedures (12%). CONCLUSIONS: Cardiac lead extraction using conventional methods has a high success rate of 86.7%. Success was significantly related to a shorter lead dwell time. Further prospective randomised trials are needed to compare traditional techniques with laser extraction both in terms of clinical outcome and cost-effectiveness. PMID- 14559134 TI - Decreased platelet activation and endothelial dysfunction after percutaneous mitral balloon valvuloplasty. AB - OBJECTIVE: This study was conducted to assess the changes in platelet activation and endothelial dysfunction in patients with mitral stenosis (MS) and sinus rhythm (SR) following percutaneous mitral balloon valvuloplasty (PMBV). BACKGROUND: Systemic thromboembolism is a serious complication in patients with valvular heart disease, and its incidence is highest in those with mitral stenosis. A hypercoagulable state has also been reported in patients with mitral stenosis and sinus rhythm. A recent study has shown that patients with previous PMBV had a lower incidence of thromboembolism. METHODS AND RESULTS: The study was conducted in 21 patients (two men, 19 women, mean age=34+/-6 years) with mitral stenosis and sinus rhythm (SR) who underwent percutaneous mitral balloon valvuloplasty and 17 healthy control subjects (two men, 15 women, mean age=33+/-6 years). Biochemical markers of platelet activity (beta thromboglobulin, BTG, and soluble P-selectin, sPsel) and endothelial dysfunction (von Willebrand Factor, vWF) were measured in both control subjects' and patients' serum samples taken immediately before PMBV and 24 h after PMBV procedure. All patients underwent successful PMBV. Significant improvement of mitral valve area, pulmonary artery pressure, mean mitral gradients, and left atrial diameter were achieved in all patients after PMBV. Compared with control subjects, patients with MS had higher plasma levels of BTG (66+/-26 ng/ml vs. 14+/-6 ng/ml, P<0.001), vWF (177+/-67 units/dl vs. 99+/-37 units/dl, P<0.0001), sPsel (226+/-74 ng/ml vs. 155+/-66 ng/ml, P<0.001). There was a significant reduction of plasma levels of BTG (66+/ 26 ng/ml vs. 48+/-20 ng/ml, P=0.002), vWF (177+/-67 units/dl vs. 134+/-60 units/dl, P=0.001) and P-selectin (226+/-74 ng/ml vs. 173+/-71 ng/ml, P=0.008,) 24 h after PMBV. CONCLUSION: We have shown that patients with severe MS and SR have increased platelet activation and endothelial dysfunction compared with control subjects and PMBV results in decreased platelet activity and improvement of endothelial injury. PMID- 14559135 TI - Beneficial effects of betaxolol, a selective antagonist of beta-1 adrenoceptors, on exercise-induced myocardial ischemia in patients with coronary vasospasm. AB - Although beta-blockers can not be used for the treatment of vasospastic angina, the effect of beta-blockers with vasorelaxant property on coronary vasospasm remains uncertain. In this study, we evaluated the effect of betaxolol, a new beta-blocker with calcium antagonistic property, as an additional therapy on vasospastic angina (VSA) with anginal attacks on effort. We enrolled 12 patients with VSA and anginal attacks with ST segment depression during exercise stress test. All patients received 1.25-5 mg of betaxolol for 3 months. Treadmill exercise stress test and adenosine triphosphate stress thallium-201 myocardial scintigraphy were performed before and 3 months after the onset of the betaxolol treatment. The other drugs including calcium antagonists, nitrates and nicorandil were continued. No patients experienced the exacerbation of angina during the betaxolol treatment. Exercise time to chest pain (317.5+/-72.1-454.2+/-75.5 s, P<0.01) and maximal ST segment depression (1.67+/-0.67-1.16+/-0.46 mm, P<0.01) obtained by exercise stress test, the defect score (8.6+/-2.7-5.3+/-2.1, P<0.01), the extent score (14.8+/-5.8-8.8+/-4.6%, P<0.01), the severity score (17.5+/-7.3 11.3+/-5.2, P<0.01) and washout rate (31.4+/-5.6-37.6+/-5.0%, P<0.01) obtained by the scintigraphy were improved by betaxolol. Our results suggest that betaxolol increases regional myocardial blood flow and improves exercise capacity in patients with VSA. Betaxolol may become a drug for a new potential therapy for VSA. PMID- 14559136 TI - Admissions with atrial fibrillation in a multiracial population in Kuala Lumpur, Malaysia. AB - BACKGROUND: There are established differences in cardiovascular disease in different racial groups. Worldwide, the literature regarding the clinical epidemiology of atrial fibrillation in non-white populations is scarce. OBJECTIVES: To document the prevalence of atrial fibrillation (AF) in the multiracial population of Malaysia, and to describe the clinical features and management of these patients. SETTING: Busy city centre general hospital in Kuala Lumpur, Malaysia, over a 1-month period. SUBJECTS: One-thousand four hundred and thirty-five acute medical admissions, of whom 40 patients (2.8%) had AF. RESULTS: Of 1435 acute medical admissions to Kuala Lumpur General Hospital over the 4-week study period, 40 had AF (21 male, 19 female; mean age 65 years). Of these, 18 were Malay, 16 Chinese and six Indian. Nineteen patients had previously known AF (seven with paroxysmal AF) and 21 were newly diagnosed cases. The principal associated medical conditions were ischaemic heart disease (42.5%), hypertension (40%) and heart failure (40%). Dyspnoea was the commonest presentation, whilst stroke was the cause of presentation in only two patients. Investigations were under-utilised, with chest X-ray and echocardiography in only 62.5% of patients and thyroid function checked in 15%. Only 16% of those with previously diagnosed AF were on warfarin, with a further three on aspirin. Anticoagulant therapy was started in 13.5% of patients previously not on warfarin, and aspirin in 8%. Records of contraindications to warfarin were unreliable, being identified in only 25%. For those with known AF, 58% were on digoxin. For new onset AF, digoxin was again the most common rate-limiting treatment, initiated in 38%, whilst five patients with new onset AF were commenced on amiodarone. DC cardioversion was not used in any of the patients with new onset AF. CONCLUSION: Amongst acute medical admissions to a single centre in Malaysia the prevalence of AF was 2.8%. Consistent with previous similar surveys in mainly western (caucasian) populations, standard investigations in this Malaysian cohort were also inadequate and there was underuse of anticoagulation, medication for ventricular rate control and cardioversion to sinus rhythm. PMID- 14559137 TI - Low molecular weight heparin (LMWH)-induced skin necrosis in a patient with unstable angina. PMID- 14559138 TI - Cardiovascular magnetic resonance of imminent cardiac tamponade due to postpericardiotomy syndrome. PMID- 14559139 TI - Rapidly fatal acute amiodarone hepatitis occurring in the context of multiple organ failure. PMID- 14559140 TI - Cardiovascular magnetic resonance of iatrogenic ventricular scarring due to catheter ablation for left ventricular tachycardia. PMID- 14559141 TI - Multidetector-row helical computed tomography imaging of unroofed coronary sinus. PMID- 14559142 TI - Brugada-type ECG with polymorphic ventricular tachycardia: a red herring for isolated right ventricular infarction. AB - We present a patient with Brugada-type ECG abnormalities and recurrent polymorphic ventricular tachycardia (VT). Subsequent investigations confirmed the diagnosis of isolated right ventricular myocardial infarction. The VT resolved after the 1st day and was not inducible subsequently. This case illustrates the importance of a careful study of the ECG to exclude other conditions in a patient with Brugada-type ECG abnormalities. PMID- 14559143 TI - Effects of a beta blocker and spironolactone on plasma homocysteine levels. PMID- 14559144 TI - Appearance of voltage-gated calcium channels following overexpression of ATPase II cDNA in neuronal HN2 cells. AB - ATPase II (a Mg2+-ATPase) is also believed to harbor aminophospholipid translocase (APTL) activity, which is responsible for the translocation of phosphatidylserine (PS) from the outer leaflet of the plasma membrane to the inner. To test this hypothesis we overexpressed the mouse ATPase II cDNA in the neuronal HN2 cells. In addition to a dramatic increase in APTL activity, we also made the unexpected observation that expression of the mouse ATPase II cDNA from the vector pCMV6 resulted in the appearance of calcium current. Although the hybrid cell line HN2 or a line (HN2V32) obtained by expressing a heterologous gene from the same expression vector showed no calcium current, both ATPase II overexpressing clones (HN2A12 and HN2A22) showed significant barium conductance. This current was due to calcium channels because it was blocked almost completely by 100 microM CdCl2 and it had a significant N-type component since it was blocked by 38.5% in the presence of 5 microM omega-conotoxin (omega-CTX). Western blot analysis using an antibody against the N-type calcium-channel alpha1B subunit revealed a dramatic increase in expression of this protein in the HN2A12 and HN2A22 cell lines. Our results suggest that ATPase II also harbors APTL activity. In view of the prior knowledge that APTL activity is inhibited by an increase in calcium, our results also suggest that APTL expression exerts a negative feedback regulation on itself by inducing expression of channels that cause an influx of calcium ions. The mechanism of this regulation could reveal important information on a possible cross-regulation between these two families of proteins in neuronal cells. PMID- 14559145 TI - Proteomic analysis of the synaptic plasma membrane fraction isolated from rat forebrain. AB - Mass spectrometry (MS) in conjunction with liquid chromatography and gel separation techniques has been utilized to identify synaptic plasma membrane (SPM) proteins isolated from rat forebrain and digested with the protease trypsin. Initial results employing two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) separation of the SPM protein mixture have shown that several membrane proteins were under-represented due to solubilization problems in the dimension of isoelectric-point focusing. Given the complexity of the SPM, multiple stages of separation were necessary prior to mass spectrometric detection in order to facilitate protein identification. This particular study involved several approaches using one-dimensional (1D) sodium dodecyl sulfate (SDS)-PAGE, strong cation-exchange (SCX) chromatography and capillary reversed phase high performance liquid chromatography (HPLC) techniques. In addition to these gel and HPLC separation stages, complementary information was obtained by using both matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) mass spectrometry. Data-dependent acquisition employing capillary HPLC-nanoESI/MS allowed for the detection of low-abundance tryptic peptides in the digested SPM fraction and identification of the corresponding proteins when product-ion information of a single or multiple peptides was used in protein database searching. The potential value of this subproteome methodology was exemplified by the identification of several proteins relevant to synaptic physiology which included various transporters, receptors, ion channels, and enzymes. PMID- 14559147 TI - Subunit-dependent effects of nickel on NMDA receptor channels. AB - Nickel (Ni2+) is a transition metal that affects different neuronal ionic channels. We investigated its effects on glutamate channels of the NMDA-type in the presence of saturating concentration of glutamate or NMDA (50 microM), in 0 external Mg and in the continuous presence of saturating glycine (30 microM). In neonatal rat cerebellar granule cells, Ni2+ inhibited the current evoked by NMDA at -60 mV with an IC50 close to 40 microM. The inhibition was weakly voltage dependent and the current at +40 mV was inhibited with IC50=86 microM. Wash out of the metal unmasked a stimulatory effect which persisted for a few seconds. In HEK293 cells transiently transfected with recombinant NR1a-NR2A receptors, Ni2+ inhibited the current elicited by glutamate with an IC50=52 microM at -60 mV and 90 microM at +40 mV. In HEK293 expressing NR1a-NR2B receptors, 0.1-100 microM Ni2+ caused a potentiation of the current, with EC50=4 microM, while with 300 microM, a voltage-dependent block became apparent (IC50=170 microM). As previously reported, the current through both classes of recombinant receptors was steeply dependent on external pH, and in both cases the protonic block had an IC50 close to pH 7.2. Application of Ni2+ showed that stimulation of NR1a-NR2B receptor channels was dependent on external pH, while voltage-independent inhibition of NR1a-NR2A was less sensitive to pH change. These results indicate that Ni2+ has multiple and complex effects on NMDA channels, which are largely dependent on the NR2 subunit. PMID- 14559146 TI - Effect of 6-hydroxydopamine on striatal GDNF and nigral GFRalpha1 and RET mRNAs in the adult rat. AB - Exogenous GDNF as well as vectors containing the gene for this trophic factor has been shown to be neuroprotective in animal models of Parkinson's disease. We therefore investigated whether changes in striatal GDNF protein and nigral mRNA levels of its co-receptors GFRalpha1 and RET occur in response to lesions of dopamine (DA) neurons and examined the temporal profile of these changes as they relate to the loss of dopaminergic markers. Rats were lesioned with 6 hydroxydopamine and sacrificed 3 h to 60 days post-infusion. DA tissue levels in the striatum and tyrosine hydroxylase immunoreactivity in the substantia nigra (SN) and ventral tegmental area (VTA) were used to determine the size of the lesions. GDNF protein was measured in the striatum using radioimmunocytochemistry. In situ hybridization was used to determine alterations in the mRNAs of RET and GFRalpha1 in the SN and VTA. We observed no persistent changes in GDNF protein in the striatum in response to 6-hydroxydopamine over the 60-day observation period, suggesting that compensatory changes in this trophic factor do not occur in response to injury. Dramatic decreases in RET and GFRalpha1 were observed in both SN and VTA that were generally correlated with the loss of TH protein and striatal DA content, strongly suggesting that these receptors are located on DA neurons and that the protective effect of GDNF reflects a direct action of the trophic factor on these neurons. PMID- 14559148 TI - Okadaic acid mediates tau phosphorylation via sustained activation of the L voltage-sensitive calcium channel. AB - Accumulation of phosphorylated isoforms of the microtubule-associated protein tau is one hallmark of affected neurons in Alzheimer's disease (AD). This increase has been attributed to increased kinase or decreased phosphatase activity. Prior studies indicate that one of the kinases that phosphorylates tau (mitogen activated protein kinase, or MAP kinase) does so at least in part indirectly within intact neuronal cells by phosphorylating and activating the L-voltage sensitive calcium channel. Resultant calcium influx then fosters tau phosphorylation via one or more calcium-activated kinases. We demonstrate herein that treatment of differentiated SH-SY-5Y human neuroblastoma with the phosphatase inhibitor okadaic acid (OA) similarly may increase tau phosphorylation via sustained activation of the L-voltage-sensitive calcium channel. OA increased phospho-tau as indicated by increased immunoreactivity towards an antibody (PHF-1) directed against paired helical filaments from AD brain. This increase was blocked by co-treatment with the channel antagonist nimodipine. OA treatment increased channel phosphorylation. The increases in calcium influx, PHF-1 immunoreactivity and channel phosphorylation were all attenuated by co-treatment with PD98059, which inhibits MAP kinase activity, suggesting that OA mediates these effects at least in part via sustained activation of MAP kinase. These findings underscore that divergent and convergent kinase and phosphatase activities regulate tau phosphorylation. PMID- 14559150 TI - Extinction of cocaine self-administration produces alterations in corticotropin releasing factor gene expression in the paraventricular nucleus of the hypothalamus. AB - The long-term effect of cocaine self-administration on corticotropin releasing factor (CRF) mRNA content in the hypothalamic CRF-containing neurons has not yet been established. The purpose of this study was to examine the time course effects of the extinction of cocaine self-administration behavior on CRF gene expression in the paraventricular nucleus of the hypothalamus (PVN) using in situ hybridization histochemistry (IHHS). Seventy-two littermate male Lewis rats were randomly assigned in triads to one of three conditions: (a) contingent intravenous self-administration of 1 mg/kg/injection of cocaine (CONT), (b) non contingent injections of either 1 mg/kg/injection of cocaine (NONCONT) or (c) saline yoked (SALINE) to the intake of the self-administering subject. The self administering rats were trained to self-administer cocaine under a fixed ratio 5 (FR5) schedule of reinforcement for a minimum of 3 weeks. After stable baseline levels of drug intake had been reached, saline was substituted for drug. Following this first extinction period, cocaine self-administration was reinstated for an additional period of 2 weeks. Immediately after cessation of the last session of cocaine self-administration (Day 0) and 1, 5 and 10 days after the second extinction period, animal brains in each triad were removed to be processed for IHHS. CRF mRNA levels in the PVN were significantly lower in the NONCONT cocaine group at Day 0 compared to CONT or SALINE groups. On Day 1, hypothalamic CRF gene expression significantly decreased in the CONT cocaine group with respect to the SALINE group, but there were no differences between the cocaine groups or among the NONCONT cocaine and SALINE groups. After 5 and 10 days of extinction, no differences were found in CRF mRNA content in the PVN between the three conditions of this study. These results suggest that, after the extinction of cocaine self-administration, changes in hypothalamic CRF gene expression are differentially affected depending upon the type of cocaine administration, and that the stages of cocaine withdrawal might not be associated with enduring changes in hypothalamic CRF mRNA levels. PMID- 14559149 TI - Tyrosine phosphorylation of Disabled-1 is essential for Reelin-stimulated activation of Akt and Src family kinases. AB - Reelin is a large secreted signaling protein that is essential for proper positioning of migratory neurons during mammalian brain development. The Reelin signal is transduced into the cell by the lipoprotein receptors VLDLR and ApoER2, leading to tyrosine phosphorylation of the associated intracellular adaptor protein Disabled-1 (Dab1). Tyrosine phosphorylation of Dab1 is essential for responding to Reelin, as knock-in mice expressing a form of Dab1 that cannot be phosphorylated on tyrosine are indistinguishable from mice lacking Reelin, Reelin receptors or Dab1. Molecular events dependent on Dab1 tyrosine phosphorylation are unknown. However, Reelin has recently been shown to activate the phosphoinositide-3-kinase (PI 3-K)-dependent kinase, Akt, as well as Src family kinases in wild type but not Dab1-/- primary embryonic neuronal cultures. Using pharmacological inhibitors and mice harboring mutant alleles of Dab1, we show here that tyrosine phosphorylation, but not the carboxyl-terminal region, of Dab1 is required for Reelin-induced activation of Akt and Src family kinases. Additionally, although Fyn is an important regulator of Dab1, Fyn deficiency does not prevent acute Reelin-induced Akt activation. Finally, whereas a number of growth factors propagate signals simultaneously through PI 3-K and mitogen activated protein kinase (MAPK) cascades, we find Reelin does not engage the canonical MAPK cascade. These results define the first molecular events strictly dependent on Reelin-induced Dab1 tyrosine phosphorylation, and suggest that propagation of the Reelin signal is mediated by Akt, substrates of Src family kinases and/or unidentified molecules that share with these a common molecular link to phosphorylated Dab1. PMID- 14559151 TI - Glutamate receptor RNA editing: a molecular analysis of GluR2, GluR5 and GluR6 in human brain tissues and in NT2 cells following in vitro neural differentiation. AB - The properties of some glutamate receptors are modified by RNA editing. This post transcriptional mechanism involves the enzymatic deamination of specific adenosines in the pre-mRNA of the glutamate receptors, performed by specific RNA adenosine deaminases (ADARs). This event gives rise to the substitution of a gene encoded amino acid with a different one that modifies the physiological properties of the ion channel. Here we report an analysis of the editing levels of AMPA GluR2, and kainate GluR5 and GluR6 in a human teratocarcinoma cell line (NT2) during in vitro neural differentiation, in conjunction with an analysis of the expression levels of GluR and ADAR genes. The editing levels were analysed using a specific standardised assay based on sequence analysis. This assay can be performed on all editing sites with a high level of sensitivity and reproducibility. Whereas GluR gene expression increased during NT2 neural differentiation, the expression of ADAR genes may be detected at comparable levels even in undifferentiated NT2 cells, remaining relatively stable during the differentiation process. Furthermore, most of the glutamate receptor editing sites increased their editing levels during NT2 neural differentiation, suggesting that the level of ADAR mRNAs is not closely related to the variable editing levels detected in the GluRs analysed. In human brain tissues, the editing levels appeared finely modulated in the different areas, indicating the possible formation of ion channels with different functional properties, thus generating a complex tissue-specific regulation of receptors and modulation of excitatory stimuli. PMID- 14559152 TI - SEPT5_v2 is a parkin-binding protein. AB - Mutations in parkin are associated with various inherited forms of Parkinson's disease (PD). Parkin is a ubiquitin ligase enzyme that catalyzes the covalent attachment of ubiquitin moieties onto substrate proteins destined for proteasomal degradation. The substrates of parkin-mediated ubiquitination have yet to be completely identified. Using a yeast two-hybrid screen, we isolated the septin, human SEPT5_v2 (also known as cell division control-related protein 2), as a putative parkin-binding protein. SEPT5_v2 is highly homologous to another septin, SEPT5, which was recently identified as a target for parkin-mediated ubiquitination. SEPT5_v2 binds to parkin at the amino terminus and in the ring finger domains. Several lines of evidence have validated the putative link between parkin and SEPT5_v2. Parkin co-precipitates with SEPT5_v2 from human substantia nigra lysates. Parkin ubiquitinates SEPT5_v2 in vitro, and both SEPT5_v1 and SEPT5_v2 accumulate in brains of patients with ARJP, suggesting that parkin is essential for the normal metabolism of these proteins. These findings suggest that an important relationship exists between parkin and septins. PMID- 14559153 TI - Over-expression of corticotropin-releasing factor mRNA in inferior olivary neurons of rolling mouse Nagoya. AB - Expression of corticotropin-releasing factor (CRF) mRNA was examined in the inferior olivary nucleus (ION) of an ataxic mutant, rolling mouse Nagoya (RMN) by semi-quantitative in situ hybridization. The most marked difference in the level of CRF mRNA signals between RMN and non-ataxic littermates (control mice) was observed in the beta-subnucleus and ventrolateral protrusion of the ION. The level of signals in these subnuclei was about twofold higher in RMN than in the controls. Signal levels in the dorsal nucleus, principal nucleus and subnucleus A were slightly but significantly higher in RMN than in the controls. In the other subnuclei, there were no differences in signal level between RMN and controls. These results suggest a region-related over-expression of CRF mRNA in the ION of RMN. This may be responsible for the increased sensitivity of some Purkinje cells to glutamate, resulting in ataxic symptoms of RMN. PMID- 14559154 TI - Changes in hippocampal gene expression after neuroprotective activation of group I metabotropic glutamate receptors. AB - Stimulation of group I metabotropic glutamate receptors (mGluRs) has been shown to protect against N-methyl-D-aspartate receptor-mediated cell death, but the underlying cellular mechanism is unknown. Using cDNA microarrays we have now compared gene expressions in organotypic hippocampal slice cultures after neuroprotective activation of group I mGluRs with (S)-3,5-dihydroxyphenylglycine (DHPG; 10 microM, 2 h) with untreated control cultures. Total RNA was extracted from the cultures immediately after the neuroprotective treatment, reverse transcribed to cDNA with incorporation of [32]P-dCTP, and then hybridized to the arrays. Of a total of 1128 genes on the Neuroarray, 33 genes displayed significant changes in expression after DHPG-treatment (six up- and 27 downregulated). These genes have been associated with regulation of synaptic excitation, inflammation, cell adhesion, cell death, and transcription. The small GTPase RAB5B associated with endocytosis emerged as a primary candidate gene for neuroprotection, and its expression was confirmed by Western blot analysis and real time polymerase chain reaction. By providing insight into genes involved in neuroprotection these data may help to identify novel therapeutic targets. PMID- 14559155 TI - Sensory neurons from mice lacking the Brn-3b POU family transcription factor are resistant to death-inducing stimuli both in vitro and in vivo. AB - The critical role for programmed cell death in the normal development of the nervous system and the abnormal cell death that occurs in human neurodegenerative diseases renders it of vital importance to identify factors that enhance or reduce the levels of cell death in specific neuronal cells in the nervous system. We show that the Brn-3b transcription factor is essential for normal cell death responses in the peripheral nervous system and that in its absence neurons are resistant to a variety of death-inducing stimuli both in vitro and in vivo. PMID- 14559156 TI - Expression and activity of poly(ADP-ribose) glycohydrolase in cultured astrocytes, neurons, and C6 glioma cells. AB - Poly(ADP-ribose) metabolism plays a major role in DNA repair, transcription, replication, and recombination. Poly(ADP-ribose) polymerases are localized primarily to the nucleus, whereas significant levels of poly(ADP-ribose) glycohydrolase (PARG) are believed to be located in the cytoplasm. Only one PARG gene has been identified, but prior studies have reported multiple products of this gene. Here we studied PARG activity and PARG gene expression in several CNS cell types that span the cell growth spectrum: rapidly dividing C6 glioma tumor cells, dividing astrocytes, non-dividing astrocytes (due to contact inhibition), and post-mitotic neurons. Activity assays showed no overall differences between these cell types, but the nuclear to cytoplasmic ratio of PARG activity was highest in C6 glioma cells and lowest in neurons. Western blotting revealed full length PARG as well as lower molecular weight PARG species in all four cell types. PMID- 14559157 TI - Autocrine action of BMP2 regulates expression of GDNF-mRNA in sciatic Schwann cells. AB - Schwann cell is a cell type that forms myelin sheath and provides trophic supports for neuronal cells by producing neurotrophic factors in both normal and traumatic situations. It was recently reported that after lesion of sciatic nerve, mRNA for glial cell line-derived neurotrophic factor (GDNF) is induced in nonneuronal cells in the nerve. However, the mechanism regulating GDNF-mRNA has remained largely unknown. In the present study, we searched for factors regulating the GDNF-mRNA expression in Schwann cells. First, we found that after transfer into explant culture as an in vitro lesion model, sciatic nerve segments began to express mRNA for bone morphogenetic protein-2 (BMP2) concomitantly with the induction of GDNF-mRNA. Treatment of the Schwann cells isolated from the sciatic nerve with combination of BMP2 and retinoic acid (RA) dramatically induced GDNF-mRNA, while BMP2 or RA alone had no effect. Furthermore, ionomycin, a calcium ionophore, which had even stronger activity on the induction of GDNF mRNA also induced also BMP2-mRNA in cultured Schwann cells. Effects of inhibitors of intracellular signaling pathways such as protein kinase C inhibitor and MAPKK inhibitor suggested that the molecular mechanism of the induction of GDNF-mRNA is distinct from that of BMP2-mRNA. These results suggest that the Schwann cell produced BMP2 plays an important role in the induction of GDNF after nerve injury in an autocrine fashion. PMID- 14559158 TI - Aplysia mollusk-derived growth factor is a mitogen with adenosine deaminase activity and is expressed in the developing central nervous system. AB - Mollusk-derived growth factor (MDGF), the first growth factor to be characterized in Aplysia, was purified and characterized and has both adenosine deaminase activity and stimulates cell proliferation in vitro. MDGF is structurally related to a new subfamily of adenosine deaminase-related growth factors that require enzymatic activity to stimulate cell proliferation, a unique property of known growth factors. We examined the expression of MDGF protein in the CNS since MDGF mRNA increased in the developing CNS, and recent data suggest that inosine is involved in neuronal reorganization and restoration of essential circuitry after CNS injury. MDGF levels transiently increased during embryonic and post metamorphic development and in the developing CNS, but was undetectable in adult CNS. No effects on morphology or neurite extension of adult Aplysia neurons were observed. PMID- 14559159 TI - MK-801 attenuates cocaine induction of c-fos and preprodynorphin mRNA levels in Fischer rats. AB - This study shows that single (15 mg/kg) or binge (3x15 mg/kg) cocaine administration increases c-fos mRNA levels, but only binge cocaine administration increases preprodynorphin mRNA levels in the caudate/putamen of Fischer rats. This increase in both c-fos and preprodynorphin mRNA levels is attenuated by the noncompetitive NMDA antagonist MK-801 (0.25 mg/kg), suggesting a preferential role for NMDA receptor co-activation in cellular events leading to cocaine induced behaviors. PMID- 14559160 TI - Acetylcholinesterase activity and molecular isoform distribution are altered after focal cerebral ischemia. AB - Biochemical changes in rat brain acetylcholinesterase (AChE) activity and molecular isoform distribution were studied using the middle cerebral artery occlusion model. After 24 h of permanent focal ischemia, AChE specific activity from infarcted cortex was found to decrease compared with contralateral cortex. Also a decrease in the minor monomeric (G1) form of AChE is response to ischemia was observed. Ischemia-induced AChE decrease was partially diminished by pre treatment with dizocilpine (MK-801), a neuroprotective NMDA receptor blocker. PMID- 14559161 TI - A role for balance of interferon-gamma and interleukin-4 production in protective immunity against Neospora caninum infection. AB - A suitable balance in the production of Th1/Th2-type cytokines has a crucial role in the control of microbial infections. We investigated cytokine production patterns and effects during Neospora caninum infection, based on two mouse models and an in vitro system. In the acute infection of N. caninum, BALB/c-background IFN-gamma-deficient mice that were sensitive to the N. caninum infection showed high levels of IL-10 production, whereas significant levels of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) production were observed in resistant wild type mice. BALB/c mice vaccinated with recombinant vaccinia virus expressing N. caninum surface protein NcSRS2 resisted parasite spread throughout the body, low levels of IFN-gamma production and high levels of IL-4 production were observed compared to unvaccinated animals. The treatment of N. caninum-infected cells with IFN-gamma or IL-10 decreased the host-cell viability in an in vitro system using mouse macrophage J774A.1 cells. On the other hand, IL-4, but not IL-10 administration, increased the viability of N. caninum-infected and IFN-gamma treated cells. In the light of the balance of Th1/Th2-type cytokine production, an IFN-gamma/IL-4 balance may have a crucial role for the control of cellular responses against the parasite invasion. PMID- 14559162 TI - Brazilian isolates of Trypanosoma (Megatrypanum) theileri: diagnosis and differentiation of isolates from cattle and water buffalo based on biological characteristics and randomly amplified DNA sequences. AB - We detected and cultivated isolates of Trypanosoma (Megatrypanum) theileri from cattle and water buffaloes in Sao Paulo state, southeastern Brazil, which were characterized by comparing morphological, growth and molecular features. Although isolates from cattle and water buffalo were morphologically indistinguishable, they differed in their growth characteristics. A dendrogram based on randomly amplified polymorphic DNA (RAPD) patterns indicated close-genetic relationships among all isolates from both species, which were all tightly clustered together and distant from Megatrypanum species from wild mammals. In addition, isolates within the T. theileri-cluster were clearly segregated into two host-associated groups. The sequence of a synapomorphic RAPD-derived DNA fragment (Tth625), which was shared by all T. theileri trypanosomes from cattle and buffalo but not detected in any of 13 other trypanosome species, was used as target for a conventional T. theileri-specific PCR assay. We also defined RAPD fragments (Tthc606 and Tthb606) that distinguished cattle from buffalo isolates. Thus, distinct growth features and genetic variability distinguished between isolates from cattle and water buffaloes of the same geographic origin, suggesting an association of these isolates with their host species. The trypanosomes from water buffalo reported here are the first T. theileri-like isolates from the Asiatic buffalo (Bubalus bubalis) to be continuously cultured and compared with cattle isolates using biological and molecular methods. PMID- 14559164 TI - Seasonal trends of Paramphistomum cervi in Tabasco, Mexico. AB - Thirty rumens were examined every month from May 1989 to April 1992 to establish seasonal trends of infection and their relationship to the climatic factors caused by Paramphistomum cervi in cattle. The cattle were chosen from three municipalities of the Sierra region in Tabasco: Jalapa, Tacotalpa and Teapa. P. cervi was found throughout the year with prevalence between 3.33 and 96.67%, and with a yearly average of 39.10%. The major period of infection was shown in Jalapa. However, the maximum mean intensity was found in Teapa. The prevalence of the infection in all three sites was similar, showing the regional trend of P. cervi infection. Cattle were infected mainly throughout the rainy and windy seasons, during summer, autumn and at the beginning of winter. PMID- 14559163 TI - The expression of RoTat 1.2 variable surface glycoprotein (VSG) in Trypanosoma evansi and T. equiperdum. AB - In order to define whether the variable antigenic type RoTat 1.2 is restricted to Trypansoma evansi and could be used as antigen in serological tests to differentiate T. evansi from Trypansoma equiperdum, the appearance of RoTat 1.2 specific antibodies in rabbits, experimentally infected with T. evansi and T. equiperdum, respectively, was analyzed. Ten strains of T. evansi and 11 strains of T. equiperdum originating from Asia, Europe, Africa and Latin America were tested. Rabbit pre-infection sera and sera of days 7, 14, 25, 35 post-infection (p.i.) were analyzed for the presence of antibodies reactive with RoTat 1.2 in immune trypanolysis, ELISA/T. evansi and CATT/T. evansi. Within the duration of the infection (maximum 35 days), all T. evansi as well as 9 out of 11 T. equiperdum infected rabbits became positive in all these tests. The rabbits infected with T. equiperdum OVI (South Africa) and BoTat 1.1 (Morocco) remained negative in the immune trypanolysis test although the latter rabbit became positive in the CATT/T. evansi and ELISA/T. evansi. On the contrary, both rabbits were positive in immune trypanolysis when tested against their respective infecting population. From these data, we conclude that most T. equiperdum strains express isoVATs of RoTat 1.2. This explains, in part, why antibody tests based on T. evansi RoTat 1.2 cannot reliably distinguish between infections caused by T. evansi and those caused by T. equiperdum unless it can be proven that most described T. equiperdum are actually misclassified T. evansi. PMID- 14559165 TI - Familial clustering of Taenia solium cysticercosis in the rural pigs of Mexico: hints of genetic determinants in innate and acquired resistance to infection. AB - In two rural villages of the state of Puebla, Mexico, where Taenia solium pig cysticercosis is highly endemic, 120 pairs of young out-bred piglets were used to assay what proved to be an effective synthetic peptide vaccine against naturally acquired cysticercosis. Because the piglets used were all sired by one of three distinct studs in many different out-bred sows, the prevalence and intensity of infection, as well as degree of protection conferred by the vaccine, could be related to each of the three stud families (A-C). The highest prevalence was found in the C family (25%), whilst the prevalence of B and A families were 21.6 and 4.4%, respectively. Familial clustering of cases was even more conspicuous in vaccinated pigs than in not-vaccinated ones: seven of the nine cysticercosis cases that occurred in the vaccinated group belonged to the C family (7/26) and two to the B family (2/23), whilst the vaccine rendered the A family totally resistant (0/71). Parasite numbers were also higher in the C family in both nai;ve and vaccinated pigs. Familial clustering of cases and of large parasite numbers in naive and vaccinated pigs hint to the relevance of their genetic background in their innate and acquired resistance to cysticercosis. PMID- 14559166 TI - Cytochrome oxidase c subunit 1 polymorphisms show significant differences in distribution between a laboratory maintained population and a field isolate of Cooperia oncophora. AB - A 474 bp fragment of the mitochondrial cytochrome oxidase c subunit 1 (cox1) of Cooperia oncophora was cloned and sequenced. The overall nucleotide diversity of the cox1 fragment varied from 0.5 to 2.0% between individuals. Two nucleotide substitutions were found within two RsaI endonuclease restriction sites and were used in a PCR-based restriction fragment length polymorphism (PCR-RFLP) assay to asses the intra-population variation of C. oncophora. Testing 816 individuals revealed the existence of three different haplotypes, having either both (type I) or only one (types II and III) RsaI site. Laboratory maintained individuals obtained at different time points after infection showed no significant difference in the distribution of the three haplotypes. Neither was there a difference in the distribution between male and female worms, confirming that the mitochondrial genome of C. oncophora is also maternally inherited. Nevertheless, there was a significant difference in the prevalence of the RsaI point mutation in the cox1 gene between the laboratory maintained population of C. oncophora and a Dutch field isolate, indicating that these RFLPs can be used to study genetic variation within or among C. oncophora populations. PMID- 14559168 TI - Distribution of horn flies on individual cows as a percentage of the total horn fly population. AB - Twenty-three mixed-breed herd cows were phenotyped for their ability to serve as a suitable host for Haematobia irritans, the horn fly. Based upon consistent observations within the lower quartile or upper quartile of individual fly counts, four cows were phenotyped as low carriers and five cows were phenotyped as high carriers of horn flies. The cows designated as low carriers consistently carried levels of flies below the economic threshold. However, during a period of fly population explosion, low carriers harbored flies well above the economic threshold. Although the number of flies counted on these low carrying cattle increased as the population increased, the relative percentage of the population that they carried changed very little. A hypothesis is proposed to explain this observation, and future studies are suggested. PMID- 14559167 TI - Experimental infection of Black Belly and INRA 401 straight and crossbred sheep with trichostrongyle nematode parasites. AB - Compared to INRA 401 lambs reared in France, Black Belly (BB) lambs reared in Guadeloupe (F.W.I.) were highly resistant to both primary and secondary experimental infection with Haemonchus contortus. To investigate this huge inter breed difference, a nucleus flock of BB was constituted, and experiments were conducted to: (i) confirm this difference in lambs born in France, (ii) check whether it was similar for Trichostrongylus colubriformis and Teladorsagia circumcincta, and (iii) find out whether this difference was age-related. Forty BB lambs, 84 F1 lambs (BB siresxINRA 401 ewes) and 88 INRA 401 lambs born in two cohorts were used in an experimental design involving three host breeds, both genders and two age-groups (3.5- and 7-month-old when first infected). The limited availability of BB lambs made the study incomplete. Infection consisted of the administration of two doses of 10,000 infective larvae of one of the nematode species, separated by an anthelmintic treatment and an interval of 1 week before the second dose was administered. Fecal egg counts (FECs) were done on Days 28 and 35 after each infection; ewe lambs of the INRA 401 and F1 breeds were necropsied, the worm burden was established, the length of the female worms measured and the eggs in utero counted. For H. contortus and T. colubriformis, the FEC was lower in the BB than in the INRA 401 lambs, and the FEC found for the F1 lambs was intermediate. For T. circumcincta, only data for the F1 and INRA 401 lambs were available, and the FECs for these breeds were the same. In all three breeds, the FEC determined after the second dose was significantly lower than that found after the first dose in the ewe lambs, but not in the ram lambs. Infecting the lambs when they were 7- instead of 3.5-month-old significantly lowered egg excretion of both parasites in the F1 (P<0.0001), but not the INRA 401 lambs. Worm numbers in the F1 and in the INRA 401 ewe lambs confirmed the FEC data, many F1 lambs being free of T. colubriformis. The H. contortus female worms were shorter and had fewer eggs in utero in the F1 than in the INRA 401 lambs. A higher proportion of T. circumcincta was at the fourth larval stage in the F1 lambs. In conclusion, the BB breed is much more resistant than INRA 401 to H. contortus, with the F1 lambs being more like the BBs after the second dose. This difference was also found for T. colubriformis and, to a lesser extent, for T. circumcincta. PMID- 14559169 TI - Recovery of live immature cyathostome larvae from the faeces of horses by Baermann technique. AB - It was demonstrated in the present study that a simple Baermann technique using disposable materials, was successful in detecting living, motile cyathostome larvae in the faeces of horses. PMID- 14559170 TI - Lymphoid tissue tropism of porcine reproductive and respiratory syndrome virus replication during persistent infection of pigs originally exposed to virus in utero. AB - The ability of porcine reproductive and respiratory syndrome virus (PRRSV) to establish a persistent infection is the principal contributing factor to the world-wide spread of the disease. Several studies have documented the course of viral infection in postnatally infected pigs; however, very little is known regarding sites of virus replication during persistent infection of pigs exposed to PRRSV in utero. In this study, virus replication and PRRSV-specific antibody were followed for several hundred days in a group of pigs derived from three sows infected at 90 days of gestation with PRRSV isolate VR-2332. Eighty-four percent of pigs were born viremic with a mortality of 54% within 21 days after birth. At approximately 60 days sera from pigs were negative for virus by virus isolation. Analysis of virus replication in the tissues of pigs randomly sacrificed between 63 and 132 days showed no evidence of virus in lung and other non-lymphoid organs. However, virus was easily recovered from tonsil and lymph nodes and in situ hybridization identified these tissues as sites of virus replication. Even though replication was at a low level, virus was easily transmitted to sentinel pigs. By 260 days pigs became seronegative and did not transmit virus to sentinel pigs. Sacrifice of remaining pigs after 300 days showed no evidence of virus in blood and tissues. This study shows that congenital PRRSV-infected pigs can support virus replication for an extended period during which virus replication is primarily restricted to tonsil and lymph nodes. PMID- 14559171 TI - Characterization of an avianpox virus isolated from an Andean condor (Vultur gryphus). AB - A novel pox virus, condorpox virus (CPV) isolated from the spleen of an Andean condor (Vultur gryphus) by inoculation of chorioallantoic membranes (CAM) of specific pathogen free (SPF) chicken embryos was compared biologically, antigenically and genetically with fowlpox virus (FPV), the type species of the genus Avipoxvirus. Susceptible chickens inoculated with CPV developed only mild localized lesions but were not protected against subsequent challenge with FPV. Based on Western blotting, in addition to the presence of cross-reacting antigens, distinct differences in antigenic profiles of CPV and FPV were observed. Sequence analysis of a 4.5 kb HindIII fragment of CPV genomic DNA revealed the presence of eight co-linear genes corresponding to FPV open reading frame (ORF)193-198, 201 and 203. Interestingly, reticuloendotheliosis virus (REV) sequences present in the genome of all FPV were absent in CPV. Although, the results of a phylogenic analysis suggested that CPV is a member of the genus Avipoxvirus, its unique antigenic, biologic and genetic characteristics distinguish it from FPV to be considered as a new member of this genus. PMID- 14559172 TI - Experimental infection of weaner sheep with S strain Mycobacterium avium subsp. paratuberculosis. AB - We sought to determine whether infection of recently weaned 12-16-week-old Merino lambs with an Australian S strain M. a. paratuberculosis, at doses consistent with natural exposure, could be detected in the first few months post inoculation. Such detection would facilitate the use of weaner sheep as sentinel animals for the presence of infectious doses of M. a. paratuberculosis on pastures. In controlled pen trials, oral doses of approximately 10(7)-10(8) viable organisms were demonstrated to be infective, whereas doses below 10(4) organisms failed to produce detectable infection. Mycobacterium avium subsp. paratuberculosis (M. a. paratuberculosis) was isolated from intestinal and/or lymphoid tissues collected at necropsy 7 or 14 weeks after first infection, but there were no associated gross or microscopic lesions. Skin testing with intradermal Johnin detected all three infected lambs at 13 weeks post-infection, and one of the three infected lambs at 6 weeks post-infection, with 100% specificity. Results for whole blood IFN-gamma assay showed some correlation with infection status but lacked specificity. One infected lamb gave a positive result in an ELISA for antibodies to M. a. paratuberculosis, 14 weeks post-infection and 1 week after skin testing. This was the first demonstration of experimental infection with S strain M. a. paratuberculosis in Australian Merino sheep at doses likely to be representative of natural infection. Culture from tissues in the first few months post-exposure could facilitate the use of naive weaner sheep as tracer animals to detect heavy contamination of pastures with M. a. paratuberculosis, but low-level contamination may not be detected in such a system. PMID- 14559173 TI - Prevalence of the lpfO113 gene cluster among Escherichia coli O157 isolates from different sources. AB - Domestic farm animals represent an important reservoir of infection for Shiga toxin-producing Escherichia coli (STEC). Nevertheless the bacterial factors required to colonise these hosts are poorly defined. In this study, the prevalence of a recently described fimbrial gene cluster, lpfO113, among human and animal isolates of STEC was investigated. lpfO113 has been shown to play a role in the adherence of STEC O113:H21 to epithelial cells. Here the presence of the lpfAO113 gene (predicted to encode a major fimbrial subunit) was examined by PCR in E. coli of serogroups O157 and O26 isolated from pigs (n=38), cattle (n=10), and humans (n=9). In addition, we tested for several other genetic virulence markers including Shiga toxin (stx), intimin (eae), the translocated intimin receptor (tir), EHEC-hemolysin (ehx) and F18 fimbriae (fedA). Overall 45 of the 57 strains (79%) possessed the lpfAO113 gene as determined by the presence of a 573 bp PCR product. Moreover, there was a close correlation between the presence of the lpfAO113 marker and the absence of the eae gene. lpfAO113 was found in all of pig isolates, suggesting a possible role in colonisation of the porcine host. In addition, several E. coli strains isolated from pigs had two fimbrial gene markers, fedA and lpfAO113. lpfAO113 was not present in strains of E. coli O157:H7 as described previously. Overall these results show that lpfAO113 is widely distributed among eae-negative E. coli isolates and thus may represent an important adherence factor in this group of pathogens. PMID- 14559174 TI - Antimicrobial resistance in Campylobacter from broilers: association with production type and antimicrobial use. AB - The isolation and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli strains from broilers arriving in French slaughterhouses, were analysed according to production types (i.e. standard, export or free-range) and antimicrobial (i.e. coccidiostats, growth promoters or therapeutic agents) administration in flocks. Prevalence was 56.6% in standard, 51.3% in export and 80.0% in free-range broilers. Three hundred and ninety-three strains were identified. Two-thirds of the strains belonged to the species C. jejuni. The others were C. coli. Antimicrobial susceptibility testing was carried out for ampicillin, nalidixic acid, enrofloxacin, tetracycline, erythromycin and gentamicin according to a dilution method. The percentages of resistant strains were, 23, 25, 17, 57, 0.3 and 0% for C. jejuni and 29, 43, 40, 70, 31 and 0% for C. coli. Statistical analysis revealed significant difference in distribution of C. jejuni and C. coli and antimicrobial resistance according to production type or antimicrobial administration. PMID- 14559175 TI - IS900/ERIC-PCR as a tool to distinguish Mycobacterium avium subsp. paratuberculosis from closely related mycobacteria. AB - There is an increasing demand for fast and reliable methods to distinguish Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) from closely related mycobacteria and also a need for rapid strain specific typing of clinical isolates for epidemiological reasons. In the present study, the potential of rep PCR as a fingerprinting method for M. paratuberculosis was assessed and compared to conventional RFLP. A PCR assay was designed and optimised to obtain reproducible fingerprints of mycobacterial DNA with primers targeting the enterobacterial intergenic consensus (ERIC) sequence and the M. paratuberculosis specific insertion sequence IS900. Reproducible fingerprints were obtained with 60 strains of M. paratuberculosis, 16 strains of M. avium subsp. avium, 3 strains of M. intracellulare, and 11 other mycobacterial strains. A species-specific band pattern that was clearly distinguishable from that of other mycobacteria was obtained with M. paratuberculosis. The rep-PCR did not detect any differences among M. paratuberculosis strains of different RFLP types, and was therefore not considered as an alternative fingerprinting method. However, the species-specific band pattern make IS900/ERIC-PCR a suitable alternative for distinguishing M. paratuberculosis from other mycobacteria, especially in cases of IS900 PCR positive mycobacteria. The fingerprinting method reported was fast and easy to perform, and produced highly reproducible results. PMID- 14559176 TI - Disseminated pythiosis in three horses. AB - Three cases of equine subcutaneous pythiosis with dissemination to the internal organs were investigated. The subcutaneous lesions were observed on the mammary gland, nostrils and limbs of the infected horses. Histopathological analysis of the infected tissues revealed a strong eosinophilic reaction, with macrophages, mast cells and giant cells. Sparsely septated hyphal filaments of 4-6 microm diameter were identified in the center of the eosinophilic areas. Specific fluorescent antibody against Pythium insidiosum confirmed the hyphae in the infected tissues in three examined horses. In one of the three cases, the DNA sequences amplified from the infected subcutaneous tissues and internal organs, revealed that P. insidiosum's 18S SSU rDNA amplicons shared 100% identity with those sequences deposit in GenBank. This is the first report confirming by immunochemical and genetic techniques that P. insidiosum can disseminated from superficial to deep structures. PMID- 14559177 TI - Ten commandments of enzymology, amended. PMID- 14559178 TI - The interaction of BMP-7 and ActRII implicates a new mode of receptor assembly. AB - The recently described structure of bone morphogenetic protein 7 in complex with the extracellular domain of the activin type receptor II provides a new and important paradigm to add to the list of possible modes of receptor assembly. A new mode of a ligand-mediated cooperative receptor assembly without receptor receptor contacts yields new and exciting insights into the molecular signal transduction mechanism in the transforming growth factor-beta superfamily. PMID- 14559180 TI - POTRA: a conserved domain in the FtsQ family and a class of beta-barrel outer membrane proteins. AB - POTRA (for polypeptide-transport-associated domain) is a novel domain identified in proteins of the ShlB, Toc75, D15 and FtsQ/DivIB families. In most cases, the POTRA domain is associated with a beta-barrel outer membrane domain and its function has been experimentally related to polypeptide transport in Toc75 (Tic Toc protein import system in chloroplast) and ShlB families. In addition to potential key roles in protein transport across the outer membrane and in bacterial septation, the POTRA domain has attractive features for vaccine development in diseases such as cholera, meningitis, gonorrhoea and syphilis. PMID- 14559179 TI - Genes within genes within bacteria. AB - Recently, an unusual gene structure has been described in species of the genus Thermus, in which the rpmH (ribosomal protein L34) coding sequence was found to be entirely overlapped by the unusually large rnpA (RNase P protein subunit) sequence. Gene overlap is common in viruses, but has not been seen to this extent in any bacterium. PMID- 14559181 TI - Tyrosine kinase signal specificity: lessons from the HGF receptor. AB - There are two major lines of thinking concerning the mechanisms responsible for specificity in receptor tyrosine kinase signalling. On one hand, receptors might provide instructive signals that dictate cell-fate decisions and, on the other, they might generate permissive signals unleashing responses that are inherently defined in the protein repertoire of target cells. Recent data indicate that the signalling activity of the Met receptor for hepatocyte growth factor is affected by association with cell-specific surface molecules, namely the alpha6beta4 integrin, Plexin B1 and CD44. This suggests that integration of cell-restricted expression of receptor partners that modulate kinase outputs with the intrinsic signalling features of receptors is required for specification of biological responses. PMID- 14559182 TI - The microRNA world: small is mighty. AB - A new paradigm of RNA-directed gene expression regulation has emerged recently, profound in scope but arresting in the apparent simplicity of its core mechanism. Cells express numerous small ( approximately 22 nucleotide) RNAs that act as specificity determinants to direct destruction or translational repression of their mRNA targets. These small RNAs arise from processing of double-stranded RNA by the Dicer nuclease and incorporate with proteins that belong to the Argonaute family. Small RNAs might also target and silence homologous DNA sequences. The immense potential of small RNAs as controllers of gene networks is just beginning to unfold. PMID- 14559183 TI - More than folding: localized functions of cytosolic chaperones. AB - Compared with other chaperone systems, heat shock proteins Hsp70 and Hsp90 interact with a larger variety of co-chaperone proteins that regulate their activity or aid in the folding of specific substrate proteins. Although many co chaperones are soluble cytosolic proteins, co-chaperone domains are also found in modular adaptor proteins, which are often localized to intracellular membranes or elements of the cytoskeleton. These specialized co-chaperones include auxilin, cysteine string protein, Tom70, UNC-45 and homologs of Bag-1. The localized co chaperones can harness the ATP-dependent mechanisms of Hsp70 and Hsp90 to do conformational work in diverse functional contexts, including vesicle secretion and recycling, protein transport and the regulated assembly and/or disassembly of protein complexes. Such flexibility is unique to the cytosolic Hsp70 and Hsp90 chaperone system. PMID- 14559184 TI - Holliday junctions in the eukaryotic nucleus: resolution in sight? AB - The Holliday junction is a key recombination intermediate whose resolution generates crossovers. Interplay between recombination, repair and replication has moved the Holliday junction to the center stage of nuclear DNA metabolism. Holliday junction resolvases in the eukaryotic nucleus have long eluded identification. The endonucleases Mus81/Mms4-Eme1 and XPF-MEI-9/MUS312 are structurally related to the archaeal resolvase Hjc and were found to be involved in crossover formation in budding yeast and flies, respectively. Although these endonucleases might represent one class of eukaryotic resolvases, their substrate preference opens up the possibility that junctions other than classical Holliday junctions might contribute to crossovers. Holliday junction resolution to non crossover products can also be achieved topologically, for example, by the action of RecQ-like DNA helicases combined with topoisomerase III. PMID- 14559185 TI - Biochemical and molecular characterization of diseases linked to motor proteins. AB - Recent studies have revealed that kinesin, dynein and myosin each form large superfamilies and participate in many different intracellular transport systems. Importantly, these motor proteins play significant roles in the pathogenesis of a variety of diseases. Studies using knockout mice for kinesin KIF1B have led to the identification of the cause of a human hereditary neuropathy, Charcot-Marie Tooth disease type 2A. The function of members of the dynein superfamily whose existence has previously only been confirmed through genome databases, has been revealed by studies of immotile cilia syndrome. Unconventional myosins have been shown to function in the inner-ear cells by examination of hereditary human hearing impairment and studies using mouse models. In addition, some diseases are caused by mutations, not in the motor itself, but in the proteins associated with the motor proteins. Here, we discuss the relationship of these motor proteins and how they contribute to disease in molecular terms. PMID- 14559186 TI - Historical review: the discovery of 'giant' RNA and RNA processing: 40 years of enigma. AB - RNA processing is a primordial paradigm of gene expression. Iconoclastic when discovered, after 40 years there is still no general rationale for this apparent 'wasting' of up to 90% of RNA transcripts. This article tells the story of the discovery of RNA in the laboratory of J.E. Darnell. The discovery of 'giant' RNA and its conversion into rRNA revealed the phenomenon of RNA processing and pre rRNA. Genuine mRNA was also identified, but the majority of DNA-like nuclear RNA was also found to be giant and unstable. In spite of early evidence, pre-mRNA processing was only accepted in 1977 when the discovery of gene fragmentation in DNA made it obvious. PMID- 14559188 TI - [Sentinel lymph node in colorectal cancer]. PMID- 14559189 TI - [Modifying the clinical practices: some congratulations, one regret and one hope]. PMID- 14559190 TI - [Laparoscopic pancreatic resections]. AB - The feasibility of laparoscopic pancreatic resection has been demonstrated. However, the real clinical benefit for the patients remains questioned. The best indication for a laparoscopic approach appears to be the resection of benign or neuro-endocrine tumors without a need for pancreato-enteric reconstruction (i.e enucleation or distal pancreatectomy). The use of the laparoscopic approach for malignant tumors still remains controversial. The benefits of minimally invasive surgery are clearly correlated with the successful management of the pancreatic stump. Pancreatic related complication rate (fistula and collection) is 15% when using pancreatic transection with a laparoscopic endostappler. PMID- 14559191 TI - [Is sentinel lymph node mapping relevant for colon cancer?: a feasibility study]. AB - BACKGROUND: Sentinel lymph node (sln) technique using blue injection is controversial for colon cancer. The aim of this study was to evaluate the feasibility and interest of sentinel node detection to identify the ultrastaging rate detecting occult nodal micrometastases missed on routine H&E examination. METHODS: During surgery blue dye was injected subserously around the tumor in 30 patients operated for a colon cancer. The first lymph nodes to turn blue were noted as sln. For each sln three examination levels were performed; if no tumor was detected by H&E examination, a cytokeratine immunohistochemistry study was performed. RESULTS: For each case, one or more sln were found (100%). The median number of lymph nodes examined and of sln found was, respectively, 23 (range 10 55) and 2 (1-4). There were 21 pN0 tumors, among which we found two cases (9%) with a micrometastasis and one case of isolated tumor cells detected, resulting in a 14% (3/21) ultrastaging for pTxN0. The sln was positive in five patients out of nine with a N+ disease. CONCLUSIONS: Sln detection was a successful technique when there was no evident lymph node involvement, no primary large lesion or no associated liver metastasis. Focused examination of the sln identified 10-20% of additional ultrastaging disease for staged pT1, 2, 3N0M0 tumor. This may have an important implication for follow-up and adjuvant treatment in future protocols. PMID- 14559192 TI - [Long-term usefulness of an information programme on practices in surgical antimicrobial prophylaxis]. AB - INTRODUCTION: In France, numerous concordant studies show that there are some discrepancies between guidelines on surgical antibiotic prophylaxis and the current practice. In a previous study, conducted in April-June 2001, we found that the rate of appropriateness of surgical antimicrobial prophylaxis was approximately 40%. An information programme was implemented and the purpose of this paper was to present the short- and long-term usefulness of this campaign. METHODS: A total of 13 pairs of surgeons/anaesthetists participated in data collection during the three periods of the study. Prescriptions were observed in order to answer to five questions. Five variables describing practices concerning antibiotic prophylaxis in surgery were compared to national recommendations (updated in 1999): did the surgical procedure require antibiotic prophylaxis and was this carried out? Was the antibiotic used appropriate? Was the timing of the first injection optimal? Was the total duration of the treatment correct? Was the dose correct? RESULTS: The overall compliance with recommendations was significantly improved during the third period (P = 0.0002). This improvement was particularly marked for antimicrobial prophylaxis duration. CONCLUSION: It seems that sequential surveillance of antimicrobial prophylaxis, including numerous surgical teams, could considerably improve the practices, if it was associated to informations that allowed physicians to appropriate the procedures. PMID- 14559193 TI - [The importance of lymph node dissection in medullary thyroid macrocarcinomas]. AB - Medullary thyroid carcinoma (MTC) is often regarded as good medium-term forecast. The 5- and 10-years survival rates are, respectively, appraised at 78-85% and 70 78%. These rates take no care, however, of the fact that 50-56% of the patients keep a pathological calcitonine (CT) level giving evidence of an evolutive disease. The treatment is based on the total thyroidectomy and cervical lymphadectomy. This treatment remains often incomplete and the results of reintervention are disappointing. AIM OF THE STUDY: About 48 patients, we wanted to demonstrate the importance of a complete lymph node dissection performed in the neck as soon as possible. PATIENTS AND METHODS: Between 1979 and 2000, 48 patients were treated for macroMTC (size >1 cm). The duration of follow-up was of 1-29 years (mean 9.3 years). The complete (central and lateral) neck dissection was initially made only in 22 cases. The selected criterion to assess the result was the normalization of the basal CT level. RESULTS: The rate of node involvement was 66.6% if the complete lymphadectomies (n = 22), the secondary neck dissections (n = 15), the incomplete (n = 10) and not made lymphadectomies (n = 2) were gathered. In case of primary or secondary complete lymphadectomies, the rate of node involvement was 81%. The 22 primary complete lymphadectomies performed in 13 patients (59%) allowed to normalize the basal CT level and among 17 (77.2%) to decrease this rate over 90%. All the incomplete neck dissection failed in case of positive nodes. CONCLUSION: The frequency of node involvement in macroMTC is about 80%. It does not have a preferential territory and the bilaterality is frequent (28-49%): that justifies a bilateral complete neck dissection. Initial surgical treatment seems essential in regard to the rate of normalization of basal CT level, which is, when a first complete lymphadectomy is done and in case of iterative surgery, respectively 59 and 26.6%. A complete lymphadectomy is still too rarely carried out: 22 times (45.8%) in our own experience and from 14 to 42% in the literature. PMID- 14559194 TI - [Laparoscopic pancreatectomy: report of 22 cases]. AB - OBJECTIVE: To evaluate results of laparoscopic pancreatectomy for benign lesions of the pancreas. Peri-operative data, surgical outcomes and techniques are presented. PATIENTS AND METHODS: Eighteen women and four men underwent laparoscopic pancreatectomy and were collected retrospectively from 1999 to 2003. RESULTS: Laparoscopic pancreatectomy was attempted in 22 patients and completed successfully in 18: 10 enucleations, three distal pancreatectomies, four left pancreatectomies and one total pancreatectomy for endocrine and cystic tumors. Left and distal pancreatectomies were performed with preservation of the spleen. Four patients were converted (one enucleation, one whipple procedure and two left pancreatectomy). There was no mortality; the post-operative morbidity included two pancreatic leaks and one case of half splenic infarction. The median length of hospital stay was 12 days. CONCLUSION: Patients appear to benefit from laparoscopic pancreatectomy for pancreatic benign tumors. PMID- 14559195 TI - [Endoscopic sphincterotomy or not for an acute biliary pancreatitis]. PMID- 14559196 TI - [Choledocal cyst associated with congenital cystic duct dilatation: report of two cases]. AB - We report two cases of a choledochal cyst associated with dilatation of the cystic duct. This unusual variant of choledochal cyst was explored by ultrasonography and MR cholangiopancreatography. PMID- 14559197 TI - [Watermelon stomach: a rare cause of iron deficiency anemia, surgically treatable; a new case with review of the literature]. AB - The authors report a new case of water-melon stomach, without portal hypertension, and responsible for a iron deficiency anemia cured by antrectomy. Water-melon stomach is a particular form of gastric antral vascular ectasia, characterized by a specific and striking endoscopic aspect. The diagnostic, histologic, pathogenic and therapeutic aspects are reviewed. PMID- 14559198 TI - [Pudendal hernia of the labia]. AB - A case of hernia is reported with a brief review of perineal hernia. PMID- 14559199 TI - [Cervical lymph node dissection and thyroid carcinoma]. AB - After an anatomic recall of the cervical lymph node compartments we describe the surgical technique of the lymph node dissection in patients with differentiated thyroid carcinoma. These lymphadenectomies should be associated with total thyroidectomy. Cervical lymph node dissection always concerns central compartment and is sometimes extended to the cervico lateral compartments. Lymphadenectomy of a compartment should be complete, reoperations leading to an important morbidity. The two main complications of cervical lymph node dissection are inferior laryngeal nerve palsy and hypoparathyroidism. PMID- 14559200 TI - [The advent of surgical gloves]. AB - The advent of surgical gloves had a double evolution. Evolution of the material: cecum of a sheep, cotton, silk, leather, rubber. Originally introduced to protect theatre staff's hands from corrosive solutions, subsequent use was to protect the patient from contamination by theatre staff. Many surgeons contributed to the evolution of the surgical gloves. The use of gloves was truly part of an evolutionary process than a discovery. The turning of surgical gloves is now incontestable, and their use more and more important. Surgical gloves must secure a crossing protections between surgeon and patient. PMID- 14559203 TI - Developmental dyslexia in different languages: language-specific or universal? AB - Most of the research on developmental dyslexia comes from English-speaking countries. However, there is accumulating evidence that learning to read English is harder than learning to read other European orthographies (Seymour, Aro, & Erskine, 2003). These findings therefore suggest the need to determine whether the main English findings concerning dyslexia can be generalized to other European orthographies, all of which have less irregular spelling-to-sound correspondences than English. To do this, we conducted a study with German- and English-speaking children (n=149) in which we investigated a number of theoretically important marker effects of the reading process. The results clearly show that the similarities between dyslexic readers using different orthographies are far bigger than their differences. That is, dyslexics in both countries exhibit a reading speed deficit, a nonword reading deficit that is greater than their word reading deficit, and an extremely slow and serial phonological decoding mechanism. These problems were of similar size across orthographies and persisted even with respect to younger readers that were at the same reading level. Both groups showed that they could process larger orthographic units. However, the use of this information to supplement grapheme phoneme decoding was not fully efficient for the English dyslexics. PMID- 14559204 TI - Is perseveration caused by inhibition failure? Evidence from preschool children's inferences about word meanings. AB - Four studies examined the relation between children's cognitive inhibition and flexibility in a lexical inference task. Children's linguistic flexibility was assessed by the Flexible Induction of Meaning (FIM) test (Deak, 2000a), which requires that children shift inferences about the meanings of several words for novel objects. In Study 1, 54 3-year-olds either were trained between blocks of problems, for a delay of 3 min, or received no training or delay. Training delays did not influence perseveration. In Study 2 (N=72 3- and 4-year-olds') novel word problems were grouped either to increase the frequency of cue switches (i.e., reduce response "set") or minimize the interval between problems about the same objects. Again, no effect was found. In Study 3, 48 3- and 4-year-olds completed 6 preliminary trials; in a high interference group these trials generated a response set to be inhibited upon the first switch to a new cue context. This group did not perseverate more than a control group. There was no association between FIM perseveration and a Stroop-like test of verbal inhibition though both were marginally related to receptive vocabulary. In study 4 (48 3- and 4-year olds), FIM was again unrelated to Stroop performance, but was related to the ability to tell whether a situation or problem is indeterminate. Thus, flexibility across semantic inferences is not influenced by timing, order, and number of pre-switch problems and is not predicted by individual differences in a test of verbal inhibition. However previously reported age and individual differences in flexible induction of word meanings are robust and related to vocabulary and logical ability. PMID- 14559205 TI - Notation to symbol: Development in children's understanding of print. AB - Three studies used the moving word task to examine children's understanding of the symbolic nature of print. A card containing a printed word is presented with two pictures, one of which is named by the word. The child is told what the card says and then asked three times what the word is. For the second question, the card is spatially adjacent to the picture it does not name, and 4-year-olds typically respond that the word has changed to name this picture. The studies examined the impact of how the move is carried out and the role of other cognitive abilities (Study 1), the influence of the source of the print and the visibility of the cards (Study 2), and the role of the matching picture in children's solutions (Study 3). In all cases, children continued to name the picture that is closest to the card, indicating an incomplete grasp of how print carries meaning. The conclusion is that children's error in this task represents a fundamental misconception about how print signifies meanings, and that prior to reading, children do not understand the symbolic function of the constituents of print. Furthermore, executive processes of representation and inhibition are identified as significant to children's solutions. PMID- 14559206 TI - Infants' perception of illusory contours in static and moving figures. AB - We investigated 3-8-month-olds' (N=62) perception of illusory contours in a Kanizsa figure by using a preferential looking technique. Previous studies suggest that this ability develops around 8 months of age. However, we hypothesized that even 3-4-month-olds could perceive illusory contours in a moving figure. To check our hypothesis, we created an illusory contour figure in which the illusory square underwent lateral movement. By rotating the elements of this figure, we created non-illusory contour figures. We found that: (1) infants preferred moving illusory contours to non-illusory contours by 3-4 months of age, and (2) only 7-8-month-olds preferred static illusory contours. Our findings demonstrate that motion information promotes infants' perception of illusory contours. Our results parallel those reported in the study of partly occluded objects (). PMID- 14559207 TI - Simultaneous screening for three mutations in the ABCB1 gene. AB - A noncoding C3435T mutation in exon 26 of the ABCB1 gene was found to be often associated with a G2677T(A) mutation in exon 21 encoding an Ala893Ser P glycoprotein and with a noncoding C1236T mutation in exon 12. We developed a Pyrosequencing screening method that simultaneously detects all three mutations. After separate PCRs for each exon, the sequences around the potentially mutated nucleotide positions were simultaneously analyzed in a multiplex assay. The method was tested with DNA from 100 volunteers. Allele frequencies of the T1236, T2677, and T3435 alleles were 44, 42, and 50%, respectively. A mutation at position 3435 occurred together with a mutation at position 2677 or 1236 in 64 and 65% of the subjects, respectively. The most frequent haplotype, with 44.4%, was not mutated at all three positions, i.e., C1236, G2677, C3435. The second most frequent haplotype, with 37.1%, was mutated at all three positions, i.e., T1236, T2677, T3435. The most frequent genotype, with 36%, was heterozygously mutated at all three positions, i.e., C/T1236, G/T2677, C/T3435. The next most frequent genotypes were a homozygous nonmutated genotype, with 20%, and a homozygous mutated genotype, with 13%. PMID- 14559208 TI - Promoter and intronic variants affect the transcriptional regulation of the human dopamine transporter gene. AB - We have attempted to identify regions involved in the transcriptional regulation of the DAT1 (HUGO approved symbol SLC6A3) gene that may harbor functional variants predisposing to several neuropsychiatric disorders by examining haplotypes of various 5' and intronic regions for their effect on expression in a dopaminergic cell line. A 1.5-fold difference in regulatory activity was observed between haplotypes of the proximal promoter/intron 1 region, representing the two previously identified 5' clades. Although we found no effect on transcription with inclusion of the 9- and 10-repeat alleles of the 3' VNTR, introns 9, 12, and 14 appear to contain enhancer elements capable of increasing expression approximately 2-fold with respect to the promoter constructs. Differences in expression were also observed between two alleles of intron 14. These results thus suggest that it may be the particular combination of polymorphisms in a haplotype across the gene that ultimately affects DAT1 gene expression. PMID- 14559209 TI - Molecular analysis of human Siglec-8 orthologs relevant to mouse eosinophils: identification of mouse orthologs of Siglec-5 (mSiglec-F) and Siglec-10 (mSiglec G). AB - We recently identified a novel human sialic acid binding immunoglobulin-like lectin, Siglec-8, using mRNA from human eosinophils. To search for a mouse Siglec (mSiglec) ortholog of Siglec-8 and other mouse Siglec paralogs, we conducted public database searches with cDNA sequences of human Siglec-5 to -10 and identified two novel mSiglecs. One has significant sequence identity to human Siglec-5 and is a splice variant of mSiglec-F. The other has greatest sequence identity to human Siglec-10 (mSiglec-G). Both mSiglecs have extracellular Ig-like domains and intracellular tyrosine-based motifs. To determine whether these mSiglecs were relevant to mouse eosinophils, RT-PCR and Northern blot analysis were performed. We detected expression of mSiglec-5 (or -F), -10, and -E mRNA in purified mouse eosinophils, but Northern blot data comparing expression in tissues from normal, IL-5 transgenic, and allergen-sensitized and -challenged mice suggest that mSiglec-10 is probably most relevant to mouse eosinophils. PMID- 14559210 TI - Discovery of a null mutation in a human trace amine receptor gene. AB - G-protein-coupled receptors (GPCRs) are important mediators of signal transduction, and mutations in GPCR-encoding genes can lead to disease states. Here we describe a null mutation in an orphan GPCR-encoding gene that is predicted to inactivate completely the encoded receptor. The TA(3) receptor is a putative member of the recently described mammalian trace amine receptor family, and it is expressed in the pituitary gland and skeletal muscle. We tested for the presence of the mutant form of TA(3) (named TA(3)-TR) in a normal population, as well as in two disease groups (ADHD and bipolar affective disorder). We found TA(3)-TR to be commonly expressed in all groups, with approximately 20% allele frequency. We did not find any statistically significant correlation between either disease and the presence of TA(3)-TR. PMID- 14559211 TI - Structural and functional genomics and evolutionary relationships in the cluster of genes encoding murine 2',5'-oligoadenylate synthetases. AB - 2',5'-Oligoadenylate synthetases (2',5'-OASs) are interferon-inducible enzymes. Some of these proteins play an important role in cellular physiology, in particular, in the innate defense mechanisms against RNA virus infections. In the present publication we report the complete genomic structure of the cluster of genes encoding mouse 2',5'-OAS, with all its transcription units, their predicted functions, and their evolutionary relationships. We found that mouse Oas2/Oas3 genes have a genomic structure similar to that of human OAS2/OAS3, while the mouse equivalent of human OAS1 is composed of eight (Oas1a to Oas1h) tandemly arranged transcription units. For all these eight genes a specific inducible promoter controls transcription. The possible functions of this family of proteins are discussed. PMID- 14559212 TI - Distinct enhancers regulate neural expression of Pax7. AB - The murine Pax7 gene has emerged as an important regulator of neural and somite development. It is expressed in discrete domains of the central nervous system, including cranial neural crest, dorsal neural tube, and mesencephalic tectum, pretectum, and base, and at the midbrain-hindbrain boundary. It is also expressed by nasal epithelia and neural crest-derived facial structures. Here, we define the 5' end of the cDNA for murine Pax7 and identify the transcriptional start site. We clarify gene structure and the murine coding sequence, and we define regions of noncoding sequence that are conserved between mice and humans. Using transgenic approaches, we identify upstream and intronic regulatory elements that confer distinct domains of neural expression in the cranial neural crest, facial mesenchyme, mesencephalon, and pontine reticular nucleus. These enhancer regions will be useful for gene expression studies and for the identification of upstream regulators of Pax7 expression. PMID- 14559213 TI - Genome sequence and splice site analysis of low-fidelity DNA polymerases H and I involved in replication of damaged DNA. AB - POLH and POLI are paralogs encoding low-fidelity, class Y, DNA polymerases involved in replication of damaged DNA in the human disease xeroderma pigmentosum variant. Analysis of genomic regions for human and mouse homologs, employing the analytic tool Genome Cryptographer, detected low-repetitive or unique regions at exons and other potential control regions, especially within intron I of human POLH. The human and mouse homologs are structurally similar, but the paralogs have undergone evolutionary divergence. The information content of splice sites for human POLH, the probability that a base would contribute to splicing, was low only for the acceptor site of exon II, which is preceded by a region of high information content that could contain sequences controlling splicing. This analysis explains previous observations of tissue-specific skipping during mRNA processing, resulting in the loss of the transcription start site in exon II, in human tissues. PMID- 14559214 TI - Construction of a gorilla fosmid library and its PCR screening system. AB - A gorilla fosmid library of 261,120 independent clones was constructed and characterized. The fosmid vector is similar to the cosmid in average insert size of ca. 40 kb but contains the F factor for replication, and it is more resistant to recombination. This clone library represents about 3.7 times coverage of the gorilla genome. A simple screening system by PCR was established, and we successfully found 9 clones that cover the entire Hox A gene cluster of the gorilla genome. This gorilla fosmid DNA library is a useful resource for comparative genomics of human and apes. PMID- 14559215 TI - A 3-nucleotide deletion in the polypyrimidine tract of intron 7 of the DFNA5 gene causes nonsyndromic hearing impairment in a Chinese family. AB - Nonsyndromic inherited hearing impairment is genetically heterogeneous. Up to now, approximately 51 autosomal dominant loci implicated in nonsyndromic forms of hearing impairment have been reported in humans and 17 causative genes have been identified. Skipping of exon 8 in the DFNA5 gene has been shown to cause hearing impairment in a Dutch family. To our knowledge, no other DFNA5 mutation has been reported in familial or sporadic hearing impairment. Here, we report another mutation in DFNA5, a CTT deletion in the polypyrimidine tract of intron 7. This mutation, just like the previously reported mutation in the Dutch family, leads to skipping of exon 8 of DFNA5. In addition, we prove the existence of a recently identified short isoform of DFNA5, but the 3-nucleotide deletion reported here seems not to affect the function of this short isoform. Because no other mutation in any other part of DFNA5 has ever been described, this finding might indicate that exon 8 of DFNA5 is indispensable for the development of hearing impairment. PMID- 14559216 TI - A simple and cost-effective method for rapid genotyping of insertion/deletion polymorphisms. AB - We developed a simple method, based on the TaqMan technology, for fast genotyping of insertion/deletion polymorphisms of known location. The genotypes of 22 CEPH individuals, previously ascertained by conventional methods, were confirmed in the new assay without manual, time-consuming, post-PCR analysis. We propose to expand the application of TaqMan probes for population screening of insertion/deletion polymorphisms in which the exact endpoints of the insertion/deletion are known. The method can be applied to polymorphisms of any size and can be used for different applications such as diagnostics, genome variation, and species identification. PMID- 14559217 TI - Use of the mutual exclusivity assumption by young word learners. AB - A critical question about early word learning is whether word learning constraints such as mutual exclusivity exist and foster early language acquisition. It is well established that children will map a novel label to a novel rather than a familiar object. Evidence for the role of mutual exclusivity in such indirect word learning has been questioned because: (1) it comes mostly from 2 and 3-year-olds and (2) the findings might be accounted for, not by children avoiding second labels, but by the novel object which creates a lexical gap children are motivated to fill. Three studies addressed these concerns by having only a familiar object visible. Fifteen to seventeen and 18-20-month-olds were selected to straddle the vocabulary spurt. In Study 1, babies saw a familiar object and an opaque bucket as a location to search. Study 2 handed babies the familiar object to play with. Study 3 eliminated an obvious location to search. On the whole, babies at both ages resisted second labels for objects and, with some qualifications, tended to search for a better referent for the novel label. Thus mutual exclusivity is in place before the onset of the naming explosion. The findings demonstrate that lexical constraints enable babies to learn words even under non-optimal conditions--when speakers are not clear and referents are not visible. The results are discussed in relation to an alternative social-pragmatic account. PMID- 14559218 TI - Models of causation and the semantics of causal verbs. AB - This research examines the relationship between the concept of CAUSE as it is characterized in psychological models of causation and the meaning of causal verbs, such as the verb cause itself. According to focal set models of causation (; ), the concept of CAUSE should be more similar to the concepts of ENABLE and PREVENT than either is to each other. According to a model based on theory of force dynamics, the force dynamic model, the concepts of CAUSE, ENABLE, and PREVENT should be roughly equally similar to one another. The relationship between these predictions and the meaning of causal verbs was examined by having participants sort causal verbs and rate them with respect to the dimensions specified by the two models. The results from five experiments indicated that the force dynamic model provides a better account of the meaning of causal verbs than do focal set models of causation. Implications for causal inference and induction are discussed. PMID- 14559219 TI - Correspondence regarding M. Sharma et al., "Antioxidant inhibits tamoxifen-DNA adducts in endometrial explant culture". PMID- 14559220 TI - The alternative translation of synaptotagmin 1 mediates the non-classical release of FGF1. AB - Although the extravesicular p40 domain of the transmembrane protein, p65 synaptotagmin (Syt) 1, is essential for the non-classical export of the signal peptide-less structure, FGF1, it was not possible to identify a specific intracellular protease responsible for the processing of p65 Syt1. Surprisingly, analysis of the p65 Syt1 coding sequence revealed the presence of two potential alternative ATG codons corresponding to Met103 and Met113 both of which were flanked by Kozak sequences. Indeed, in vitro translation of a Met103Ile but not a Met113Ile p65 Syt1 point mutant exhibited reduced expression of p40 Syt1 and the double p65 Syt1 Met103Ile and Met113Ile point mutant was unable to translate the p40 Syt1 isoform. Since the expression of the p65 Syt1 double point mutant inhibited the stress-induced release of FGF1, it is likely that the alternative translation of the p65 Syt1 transcript at Met103 may be involved in the generation of intracellular p40 Syt1, a critical component of the FGF1 release pathway. PMID- 14559222 TI - The ekeko mutant demonstrates a role for tetraspanin-like protein in plant development. AB - A new mutant with disturbed cell differentiation and severely altered plant morphology was obtained by visual screening of a T-DNA mutagenized population of Arabidopsis thaliana. The T-DNA in this mutant was inserted in an unknown gene (ORF At5g46700) located on chromosome V. This gene and additional 12 genes in the Arabidopsis genome show structural homologies to a class of abundantly expressed mammalian proteins with four transmembrane domains (TM4) called tetraspanins. In animals tetraspanins are involved in different cell functions like cell development, adhesion, motility, and differentiation, probably by organizing other proteins into a network of multimolecular membrane microdomains, called the tetraspanin web. So far no function for tetraspanins in plants has been described. Here, we show that a mutation in the TM4-like gene EKEKO results in severe developmental defects that could be the result of incorrect regulation of cell differentiation. PMID- 14559221 TI - E2F decoy oligodeoxynucleotides effectively inhibit growth of human tumor cells. AB - Abnormal cell proliferation, largely dependent upon deregulation of cell-cycle regulatory proteins, is an important feature of several forms of human cancer. The transcription factor, E2F, plays a critical role in the trans-activation of several genes involved in cell-cycle regulation, thereby regulating cell growth. We have demonstrated that E2F decoy oligodeoxynucleotides (ODNs) with a circular dumbbell structure (CD-E2F decoy) corresponding to E2F binding sites effectively inhibit cell proliferation of primary cultured cells. Here we found that the E2F decoy ODNs inhibited serum-induced promoter activity of E2F-dependent genes in a sequence-specific manner in a RB-positive human osteosarcoma, U2OS, as well as in a RB-negative human cervical carcinoma, C33A. This E2F decoy ODN strongly inhibited gene expression of endogenous E2F1 and PCNA and proliferation of these cancer cells. Our results suggest that this decoy ODN strategy could represent a powerful investigative and potentially therapeutic strategy in the prevention and treatment of cancer. PMID- 14559223 TI - Introduction of short interfering RNA to silence endogenous E-selectin in vascular endothelium leads to successful inhibition of leukocyte adhesion. AB - Short interfering RNAs (siRNAs) are powerful sequence-specific reagents that suppress gene expression in mammalian cells. We report for the first time that gene silencing of endothelial E-selectin by siRNAs leads to successful inhibition of leukocyte-endothelial interaction under flow. siRNAs designed to target human E-selectin were tranfected into human umbilical vein endothelial cells (HUVEC). Western blotting analysis revealed that transfection of these siRNAs, but not the scrambled control siRNA (100nM each), attenuated E-selectin expression in HUVEC activated with TNF-alpha (10ng/ml, 4h) without affecting expression of ICAM-1. Moreover, a leukocyte adhesion assay under flow (shear stress=1.0dyne/cm(2)) demonstrated that HUVEC transfected with a siRNA against E-selectin (siE-01) supported significantly less HL60 adhesion as compared to those transfected with the control siRNA (scE-01) after activation (p<0.03). This technique provides a powerful strategy to dissect a specific function of a given molecule in leukocyte endothelial interaction. PMID- 14559224 TI - Chemical modification of contractile 3-nm-diameter filaments in Vorticella spasmoneme by diethyl-pyrocarbonate and its reversible renaturation by hydroxylamine. AB - A peritrich ciliate possesses a zooid and a long stalk consisting of a bundle of 3-nm-diameter filaments. Glycerinated stalks can contract in the presence of free Ca(2+) and re-extend in the absence of free Ca(2+). In the present study, we demonstrated that histidine residue(s) played a critical role in spasmoneme contraction by using glycerinated stalk of Vorticella. Concentration-dependent inhibition of spasmoneme contraction was observed in the presence of reversible histidine-modifying reagent named diethyl-pyrocarbonate (DEPC). In addition, the contractility degree of DEPC-modified spasmoneme could be partially restored by hydroxylamine treatment. The 244nm absorption of modified spasmoneme protein(s) increased with rising DEPC concentration and decreased following the addition of hydroxylamine treatment. Adding Ca(2+) before DEPC modification could prevent the spasmoneme contraction from inhibition of DEPC. Those results suggested that histidine residues were actively involved in spasmoneme contraction. Ca(2+) binding ability of spasmin was not inhibited by DEPC modification, which suggested that the essential histidine residues were not on the calcium-binding site of spasmin. PMID- 14559225 TI - Diverse environmental cues transiently regulate OsOPR1 of the "octadecanoid pathway" revealing its importance in rice defense/stress and development. AB - Previously, we reported rice (Oryza sativa L. japonica type cv. Nipponbare) allene oxide synthase (OsAOS) and cyclase (OsAOC) genes of the octadecanoid pathway. Here we have isolated a 12-oxo-phytodienoic acid reductase gene, called OsOPR1, encoding the last committed enzymatic step on the octadecanoid pathway leading to jasmonic acid (JA) biosynthesis. OsOPR1 encodes a 380 amino acid long polypeptide with a predicted molecular mass of 42465.02 and pI of 5.79, and belongs to a gene family in the rice genome. Transcriptional profiling using our established two-week-old rice seedling model system against a variety of environmental factors such as wounding, global signals (including JA), osmotic stresses, heavy metals, UV-C irradiation, fungal elicitor, protein phosphatase inhibitors, and gaseous pollutants indicated that OsOPR1 transcript was rapidly, transiently and differentially up-regulated within 30 min in leaves. Surprisingly, co-application of signaling molecules JA, salicylate and ethylene, resulted in a massive accumulation of the OsOPR1 transcript at 30 min and remained elevated with time, a new observation. Furthermore, transient expression of OsOPR1, most likely regulated by a de novo synthesized negative trans-acting factor(s), was evidenced by the use of cycloheximide. Finally, the endogenous OsOPR1 expression varied with the stage of plant development. These results strongly suggest a regulatory role for OsOPR1 in rice plant defense/stress response pathway(s) and reproduction. PMID- 14559226 TI - Analysis of the full-length integrase-DNA complex by a modified approach for DNA docking. AB - A model of the full-length HIV-1 integrase dimer was constructed assembling the experimentally determined structures of the single domains. Subsequently, the three-domain protein-viral DNA complex was generated for the first time through an automated docking algorithm, obtained modifying the ESCHER program, a well known method for protein-protein docking. A detailed study of the contacts established with DNA by the enzyme revealed that the predicted model reproduced the results of mutagenesis and cross-linking experiments, confirming the validity of our docking approach in predicting the base specificity in the DNA-protein interaction. PMID- 14559227 TI - Specific gene silencing using small interfering RNAs in fish embryos. AB - Recently, small interfering RNAs (siRNAs) have been used for gene knockdown in mammalian cultured cells, but their utility in fish has remained unexplored. Here we demonstrate a siRNA-mediated gene silencing technique in rainbow trout embryos. We found that siRNAs effectively suppressed the transient expression of episomally located foreign GFP genes at an early developmental stage and inhibited the expression of GFP genes in stable transgenic trout embryos. Similar gene silencing was observed with an siRNA against the endogenous tyrosinase A gene. siRNAs interfered with the expression of maternally inherited mRNA. siRNAs did not affect non-relevant gene expression and siRNAs with a 4 base mismatch did not affect target gene expression. siRNA gene silencing is therefore highly sequence-specific. Our findings are the first evidence that siRNA-mediated gene silencing is effective in fish. This technique could be a powerful tool for studying gene function during embryonic development in aquacultural fish species, zebrafish, and medaka. PMID- 14559228 TI - The impact of single cysteine residue mutations on the replication terminator protein. AB - We report the structural and biophysical consequences of cysteine substitutions in the DNA-binding replication terminator protein (RTP) of Bacillus subtilis, that resulted in an optimised RTP mutant suitable for structural studies. The cysteine residue 110 was replaced with alanine, valine or serine. Protein secondary structure and stability (using circular dichroism spectropolarimetry), self-association (using analytical ultracentrifugation), and DNA-binding measurements revealed RTP.C110S to be the most similar mutant to wild-type RTP. The C110A and C110V.RTP mutants were less soluble, less stable and showed lower DNA-binding affinity. The structure of RTP.C110S, solved to 2.5A resolution using crystallographic methods, showed no major structural perturbation due to the mutation. Heteronuclear NMR spectroscopic studies revealed subtle differences in the electronic environment about the site of mutation. The study demonstrates the suitability of serine as a substitute for cysteine in RTP and the high sensitivity of protein behaviour to single amino acid substitutions. PMID- 14559229 TI - Liposomes for microcompartmentation of enzymes and their influence on catalytic activity. AB - Modular systems for protein coupling have been applied for anchoring enzyme molecules on liposome surfaces. Two cytoplasmic model enzymes, alpha-amylase from Escherichia coli (EC. 3.2.1.1) and guanylate kinase from Saccharomyces cerevisiae (EC. 2.7.4.8), were directly coupled by a histidine-tag or indirectly via strep tag and streptavidin or streptactin linker to a liposome membrane. Though the catalytic properties of the enzymes are generally maintained, stability and specific activity of the enzymes are modified after coupling and are especially influenced by the lipid used for the liposome assembly. PMID- 14559230 TI - Optimized, highly efficient transfer of foreign genes into newborn mouse hearts in vivo. AB - Expression of foreign genes in vivo is a standard method to disclose functions of specific genes and to alter physiological conditions in distinct cell types and tissues. Virus-mediated gene transfer has proved to be a valuable tool for directed gene expression in vivo complementary to transgenic approaches. However, several problems associated with routes of application, endurance of gene expression, and efficiency of infections still have to be solved. We have optimized a gene transfer protocol into hearts of newborn mice to achieve widespread long-lasting expression using adenoviral vectors. Intrathoracic injection of high-titer adenoviral preparations (10(8)pfu) led to expression of foreign genes in >71+/-8% of all heart cells for >50 days after infection without any morphological signs of cardiac malfunction, inflammation, or immune response. This approach might be adapted to long-term cellular studies in vivo since 5 months after infection up to 20% of all cardiac cells still expressed virally encoded genes. Successful and efficient expression of other gene of interest can be easily controlled by co-injection of low titers of a reporter vector encoding EGFP (10(6)pfu). PMID- 14559231 TI - Interaction of Smads with collagen types I, III, and V. AB - Ontogenesis of the mammalian orofacial region is controlled by numerous developmental signals, including those initiated by the transforming growth factors beta (TGFbetas). Targeted deletion of the genes encoding several of the TGFbetas in mice has been shown to result in clefts of the secondary palate. Members of the TGFbeta family of growth factors utilize intracellular Smads as signal transducers. Smads 2 and 3 are transcriptional regulators that bind DNA through their conserved MH1 domains and activate/inhibit transcription of TGFbeta responsive genes through their MH2 domains. Using a yeast two-hybrid screen of a cDNA expression library constructed from fetal murine orofacial tissue, we have identified three types of collagens (types I, III, and V) that are capable of binding to the MH2 domain of Smad 3. These interactions were confirmed by glutathione S-transferase (GST) pull-down assays in which the MH2 domain of Smad 3 fused to GST interacted strongly with in vitro translated, 35S-labeled collagen types I, III, and V. Each collagen also bound to the MH2 domains of Smads 4 and 7 and, to a lesser extent, full-length Smads 1, 2, 3, and 4. Binding of Smads to collagen is a novel observation. Moreover, TGFbeta is a potent regulator of collagen synthesis and turnover during mammalian orofacial development. These data thus suggest an important means of feedback regulation of the TGFbeta signaling cascade. PMID- 14559232 TI - Role of PI3K/AKT/mTOR signaling in the cell cycle progression of human prostate cancer. AB - Prostate cancer is one of the most common cancers among men. Recent studies demonstrated that PI3K signaling is an important intracellular mediator which is involved in multiple cellular functions including proliferation, differentiation, anti-apoptosis, tumorigenesis, and angiogenesis. In the present study, we demonstrate that the inhibition of PI3K activity by LY294002, inhibited prostate cancer cell proliferation and induced the G(1) cell cycle arrest. This effect was accompanied by the decreased expression of G(1)-associated proteins including cyclin D1, CDK4, and Rb phosphorylation at Ser780, Ser795, and Ser807/811, whereas expression of CDK6 and beta-actin was not affected by LY294002. The expression of cyclin kinase inhibitor, p21(CIP1/WAF1), was induced by LY294002, while levels of p16(INK4) were decreased in the same experiment. The inhibition of PI3K activity also inhibited the phosphorylation and p70(S6K), but not MAPK. PI3K regulates cell cycle through AKT, mTOR to p70(S6K). The mTOR inhibitor rapamycin has similar inhibitory effects on G(1) cell cycle progression and expression of cyclin D1, CDK4, and Rb phosphorylation. These results suggest that PI3K mediates G(1) cell cycle progression and cyclin expression through the activation of AKT/mTOR/p70(S6K) signaling pathway in the prostate cancer cells. PMID- 14559233 TI - Self-stabilized CpG DNAs optimally activate human B cells and plasmacytoid dendritic cells. AB - We recently showed that 5'-terminal secondary structures in CpG DNA affect activity significantly more than those at the 3'-end [Biochem. Biophys. Res. Commun. 306 (2003) 948]. The need for an accessible 5'-end of CpG DNA for activity suggested that the receptor reads the DNA sequence from this end. In continuation of these studies, we have designed immunomodulatory oligonucleotides (IMOs), consisting of a nine-mer stimulatory domain, containing a CpG motif and a hairpin-loop structure at the 3'-end, referred to as self-stabilized CpG DNAs. We studied the ability of self-stabilized CpG DNAs to stimulate human B-cell proliferation and interferon-alpha (IFN-alpha) secretion in plasmacytoid dendritic cell (pDC) culture assays. Self-stabilized CpG DNAs activated human B cells and induced plasmacytoid dendritic cells to secrete high levels of IFN alpha. While both stimulatory and secondary structures in CpG DNAs were required for pDC activation, CpG motifs were sufficient to activate B cells. Interestingly, CpG motifs were not required for activity in the hairpin duplex region. Further modifications of the hairpin duplex region with a mixture of oligodeoxynucleotides and oligo-2'-O-methylribonucleotides in a heteroduplex formation permitted activation of both human B cells and pDCs. PMID- 14559234 TI - Slo2 sodium-activated K+ channels bind to the PDZ domain of PSD-95. AB - Slo2 channels are a type of sodium-activated K+ channels and possess a typical PDZ binding motif at the carboxy-terminal end. Thus, we investigated whether Slo2 channels bind to PSD-95, because it is well known that other types of K+ channels, voltage-gated and inward rectifier K+ channels, bind to PSD-95 via the PDZ binding motif and are involved in excitatory synaptic transmission. By using an extract prepared from cultured neocortical neurons, we demonstrated a biochemical interaction between mSlo2 channels and PSD-95, and a mutational analysis revealed that mSlo2 channels bound to the first PDZ domain of PSD-95 via the PDZ binding motif. To investigate the expression of mSlo2 protein in primary neocortical neurons, we raised anti-mSlo2 channel antibody and immunostained neocortical neurons. The immunocytochemical study showed that mSlo2 channels partly colocalized with PSD-95 in mouse neocortical neurons. PMID- 14559235 TI - The introduction of a phytase gene from Bacillus subtilis improved the growth performance of transgenic tobacco. AB - Phytate, the main form of phosphorus storage in plant seeds, is well known to be an anti-nutrient and a major source of phosphorus pollution in animal manure. To improve phosphorus bio-availability, we introduced a recently characterized phytase from Bacillus subtilis into the cytoplasm of tobacco cells. Although the introduction of acid fungal phytase from Aspergillus niger in previous studies did not result in any phenotypic changes in tobacco, here we show that a tobacco line transformed with a neutral phytase exhibited phenotypic changes in flowering, seed development, and response to phosphate deficiency. The transgenic line showed an increase in flower and fruit numbers, small seed syndrome, lower seed IP6/IP5 ratio, and enhanced growth under phosphate-starvation conditions compared with the wildtype. The results suggest that the over-expression of Bacillus phytase in the cytoplasm of tobacco cells shifts the equilibrium of the inositol phosphate biosynthesis pathway, thereby making more phosphate available for primary metabolism. The approach presented here can be applied as a strategy for boosting productivity in agriculture and horticulture. PMID- 14559236 TI - Sperm motility-activating complex formed by t-complex distorters. AB - Transmission ratio distortion is a dramatic example of non-Mendelian transmission. In mice, t-haplotype males produce dysfunctional +-sperm and normal t-sperm, leading to transmission in favor of t-sperm. Genetic studies have indicated that the t-complex responder locus, Tcr, rescues t-sperm but not + sperm from defective products of t-complex distorter loci, Tcds. Light chain 1 (LC1) and LC3 from sea urchin sperm outer arm dynein have sequence similarities to Tctex2 and Tctex1, respectively, both of which are wild-type products of Tcds. We show here that LC1 and LC3 are able to make a 1:1 complex. Since Tcr is a member of the Smok (sperm motility kinase) family and LC1 is phosphorylated at the activation of sperm motility in a cAMP-dependent manner, this complex in a dynein motor molecule might be a direct target of Smok/Tcr kinase in a signal cascade that regulates sperm motility. Thus, we designate it as Smoac (sperm motility activating complex). PMID- 14559237 TI - A BMP-4-dependent transcriptional control element in the 5' flanking region of Xenopus SCL gene. AB - We isolated 5.5kb genomic DNA fragment of Xenopus stem cell leukemia (SCL) that contains approximately 1.5kb of the 5' flanking region and 4.0kb of the first intron between a non-coding exon (exon 1) and a coding exon (exon 2). Sequencing result of the 5' flanking region has shown that there is a portion that shares 85% and 69% with the sequences of avian and mammalian genomes of SCL promoter region (-64 to +73). The 1.5kb 5' flanking region of SCL genome and various deletion constructs were inserted at the upstream of luciferase (luc) gene and used for the reporter assay. The reporter activity was first detected at the neurula stage in the embryos injected with -167+157/luc at the 2-cell stage and the values increased as the stages advanced. The experiments using dominant negative constructs revealed that the activation of SCL transcription via the 5' flanking region requires the BMP-4 and GATA factors. Taken together with the in situ hybridization analysis indicating that expression of SCL was downregulated in the central nervous system in BMP-depleted embryos, the proximal sequence of SCL consists of a stage-dependent and BMP signaling-dependent control element. PMID- 14559238 TI - Rapid and preferential induction of ATF3 transcription in response to low doses of UVA light. AB - Long-wavelength UV light (UVA) is known to induce transcription of various genes in the cell and to cause a variety of pathological or protective responses in the skin. To find additional UVA-responsive genes, human skin-derived fibroblasts were exposed to UVA under non- or partially lethal conditions, and the effects of UVA on the transcriptional profile were examined by using DNA microarray and RT PCR. Transcription of several genes including those already known to be UVA responsive was induced to a significant extent under 50% lethal conditions of exposure. Among those, ATF3 was the most sensitive and its transcription was increased 10-fold within 1h. Even at a non-lethal dose of UVA (8J/cm(2)), it was increased 8-fold, if cells were cultured for 3h post-exposure. Typical immediate early genes such as c-fos and c-jun were not affected at this dose. We thus suggest that ATF3 could be a key regulator for a variety of cellular responses in the skin, particularly to low doses of UVA. PMID- 14559239 TI - Gcn4 negatively regulates expression of genes subjected to nitrogen catabolite repression. AB - It has been considered that three key elements participate in nitrogen catabolite repression (NCR) of Saccharomyces cerevisiae: the GLN3 and GAT1/NIL1-encoded transcriptional activators and their negative regulator Ure2. The fact that expression of various NCR-sensitive genes is not derepressed in the absence of Ure2 has led to the proposition that there must exist a protein with a similar function to that of Ure2. The results presented in this paper show that various NCR-sensitive genes are derepressed through GLN3-mediated transcriptional activation in a gcn4Delta mutant. This effect is additive to that exerted by the lack of Ure2 and to that evoked in rapamycin-treated cultures. Our results uncover the fact that NCR is not solely achieved through the action of Gln3, Gat1, and Ure2. Since Gcn4 regulates the expression of a broad spectrum of genes, the lack of this transcriptional activator could prevent the expression of a potential Gln3 antagonist. Alternatively, Gcn4 could directly hinder Gln3 functioning. PMID- 14559240 TI - Identification of anti-TNFalpha peptides with consensus sequence. AB - Phage displayed peptide library was used to select tumor necrosis factor alpha (TNFalpha) binding peptides. After three sequential rounds of biopanning, some linear TNFalpha-binding peptides were identified from a 12-mer peptide library. A consensus sequence (L/M)HEL(Y/F)(L/M)X(W/Y/F), where X might be variable residue, was deduced from sequences of these peptides. The phages bearing these peptides showed specific binding to immobilized TNFalpha, with over 80% of phages bound being competitively eluted by free TNFalpha. To confirm the binding activity and to explore further functional properties, three peptides with typical structure were selected and expressed as GST-fused protein. These recombinant peptides effectively competed for [125I]TNFalpha binding to TNFR1 in a dose-dependent manner, with IC(50) from 10 to 160 microM. Furthermore, the GST-fused derivatives showed inhibitory effects on TNFalpha-induced cytotoxicity. Taken together, these data demonstrate that the TNFalpha-binding peptides are effective antagonists of TNFalpha and the deduced motif might be useful in development of novel low molecular weight anti-TNFalpha drugs. PMID- 14559241 TI - SOCS-3 inhibits IL-12-induced STAT4 activation by binding through its SH2 domain to the STAT4 docking site in the IL-12 receptor beta2 subunit. AB - IL-12 promotes the proliferation of T cells as well as NK cells and plays a critical role in induction of the Th1 differentiation. IL-12 mediates its biological activities through activation of the receptor-associated JAK family kinases and STAT4, which is recruited to phosphorylated Tyr-800 in the human IL 12 receptor beta2 subunit (IL-12Rbeta2). Here we demonstrate that suppressor of cytokine signaling-3 (SOCS-3) is also recruited to IL-12Rbeta2 by the interaction involving the SOCS-3 SH2 domain and phosphorylated Tyr-800 in IL-12Rbeta2. Furthermore, SOCS-3, but not its SH2 domain-defective mutant, inhibited the IL-12 induced activation of DNA-binding and transcriptional activities of STAT4. These results suggest that SOCS-3, expressed at high levels in Th2 cells, plays an inhibitory role in STAT4-mediated IL-12 signaling by binding to the STAT4 docking site in IL-12Rbeta2, thus raising a possibility that SOCS-3 may play a role in regulation of Th differentiation. PMID- 14559242 TI - Inhibitory action of telithromycin against Shiga toxin and endotoxin. AB - Shiga toxin (Stx)-producing Escherichia coli (STEC) is associated with hemolytic uremic syndrome (HUS). High inflammatory cytokine [interleukin (IL)-6 and IL-8] levels and low anti-inflammatory cytokine (IL-10) levels are indicators of a high risk for developing HUS in STEC-infected children. In this study, we investigated inhibitory action of telithromycin, a ketolide, against STEC and against Stx and lipopolysaccharide (LPS). Telithromycin inhibited in vitro STEC growth without inducing Stx phage, in marked contrast to norfloxacin. Stx markedly induced inflammatory (but not anti-inflammatory) cytokine production in human peripheral blood monocytes, while LPS induced both inflammatory and anti-inflammatory cytokine production. Telithromycin selectively inhibited the IL-6 and IL-8 production from Stx-stimulated (but not LPS-stimulated) monocytes. The drug did not significantly inhibit IL-10 production. Our data suggest that Stx plays a crucial role in the stimulation of inflammatory cytokines and such inflammatory response is inhibited by telithromycin, an anti-bacterial agent. PMID- 14559243 TI - Association of the calpain/calpastatin network with subcellular organelles. AB - The calcium-activated cysteine protease calpain is intimately involved in modulating cell adhesion and migration. The two ubiquitous isoforms of this protease, calpain I and II, are considered to be cytosolic proteins that can translocate to both focal complexes/adhesions or the plasma membrane. Using confocal microscopy and isopycnic density centrifugation, the results demonstrate that calpain I and II, the 30kDa regulatory subunit, and calpastatin associate with the endoplasmic reticulum and Golgi apparatus. Confocal microscopy reveals that calpain II colocalizes with the subcellular proteins calnexin and Rab6 in cells bound to laminin. To further verify this association, cell lysates prepared from laminin stimulated and unstimulated cells were subjected to isopycnic density centrifugation. The results reveal an increased association of calpain I, II, calpastatin, and the 30kDa regulatory subunit with the endoplasmic reticulum and Golgi apparatus as evidenced by their position in the gradient relative to calnexin, Rab6, caveolin, and beta1 integrin after laminin stimulation. This correlates with the accumulation of inducible calpain activity at the endoplasmic reticulum-Golgi apparatus interface. Further experiments established that calpain II colocalizes with phosphatidylinositol 4,5-bisphosphate. Finally, calpain II associates with membrane lipid rafts. These results provide new insights into how the calpain/calpastatin network is spatially and temporally regulated in cells binding to the extracellular matrix. PMID- 14559244 TI - Localization of NBC-1 variants in human kidney and renal cell carcinoma. AB - The Na(+)-HCO(3)(-) cotransporter (NBC-1) plays a major role in bicarbonate absorption from proximal tubules. However, which NBC-1 variant mediates proximal bicarbonate absorption has not been definitely determined. Moreover, the localization of this cotransporter in human kidney and renal cell carcinoma (RCC) tissues has not been clarified. To clarify these issues, immunohistochemical analysis was performed using the specific antibodies against kidney type (kNBC-1) and pancreatic type (pNBC-1) transporters. In Western blot analysis the expression of kNBC-1 but not of pNBC-1 was detected in both normal human kidney and RCC tissues. In immunofluorescence analysis on normal renal tissues the anti kNBC-1 antibody strongly and exclusively labeled the basolateral membranes of proximal tubules, which was confirmed by electron microscopic observation. In RCC cells, the anti-kNBC-1 antibody labeled both plasma membranes and intracellular organelles. The labeling by anti-pNBC-1 antibody was not detected in both normal kidney and RCC tissues. These results indicate that kNBC-1 is the dominant variant that mediates bicarbonate absorption from human renal proximal tubules. They also suggest that NBC-1 may have distinct roles in cancer cells. PMID- 14559245 TI - Targeted disruption of the neurochondrin/norbin gene results in embryonic lethality. AB - Neurochondrin/norbin is a cytoplasmic protein involved in dendrite outgrowth. The expression of the gene has been restricted to neural, bone, and chondral tissues. To identify the functions of the gene in vivo, we have generated mice with a disrupted mutation in the neurochondrin/norbin gene. Histological analysis of heterozygous mutant mice indicates the possibility of specific functions of neurochondrin/norbin in chondrocyte differentiation. We defined the expression patterns of neurochondrin/norbin-lacZ fusion protein in the central nervous system. In the developing olfactory bulb, beta-galactosidase activity was detected in the mantle layer at 12.5 dpc and the strongest activity was detected in the presumptive mitral or tufted cell layer at 15.5 dpc. beta-Galactosidase activity was also detected in the lateral choroid plexus. In homozygous (-/-) mutant mice, the disruption of the neurochondrin/norbin gene leads to early embryonic death between 3.5 and 6.5 dpc. This result indicates that neurochondrin/norbin gene function is essential for the early embryogenesis. PMID- 14559246 TI - ACE inhibition actively promotes cell survival by altering gene expression. AB - We tested the effect of ACE inhibition on the survival of bovine retinal (REC) and choroidal (CEC) endothelial cells (EC) in culture. The ACE inhibitor captopril delayed the apoptotic tube collapse of REC on Matrigel for >15 days. Captopril treatment of confluent monolayers (2-8 weeks) followed by slow starvation (2-4 weeks) increased EC viability by approximately 200%. Two-week captopril exposures were sufficient to confer maximal protection. Only vehicle treated EC demonstrated apoptotic features such as membrane blebbing and DNA laddering. By RT-PCR, the starvation marker p202 was upregulated only in starved cells. In REC, captopril upregulated the pro-survival proteins mortalin-2, uPA, and uPAR while downregulating the anti-growth sprouty-4 and tPA. In CEC, captopril also upregulated tPA and its inhibitor PAI-1. Amiloride (uPA inhibitor) blocked the captopril-induced increase in EC survival, secondary sprouting, and invasion in Matrigel. The pro-survival effects of captopril involve the reprogramming of genes involved in cell survival and immortalization. PMID- 14559247 TI - 9.1C3 is identical to LAIR-1, which is expressed on hematopoietic progenitors. AB - The leukocyte-associated Ig-like receptor-1 (LAIR-1) is a negative regulator of natural killer (NK) cells, its encoding gene belonging to the leukocyte receptor complex (LRC). Antibody to LAIR-1 can inhibit Ab-induced redirected lysis and TNF alpha release of effector cells. LAIR-1 contains 2 immunoreceptor tyrosine-based inhibitory motifs (ITIM) in its cytoplasmic region that have been shown to bind constitutively and presumably regulate the tyrosine phosphatase SHP-1 in hematopoietic cells. SHP-1 mutation in mice results in abnormal lymphoproliferation, suggesting that LAIR-1 may also be implicated in regulating hematopoiesis. Here we investigated a monoclonal antibody, 9.1C3, against a NK cell antigen previously described as inducing increased colony formation in in vitro assays of human bone marrow cells. We found that 9.1C3 was expressed on CD34 positive hematopoietic progenitors for the first time. In functional assays, 9.1C3 MAb was able to inhibit Ab-induced redirected lysis and TNF-alpha secretion of NK cells. We proved that 9.1C3 is identical to LAIR-1, based on the fact that not only the antigen precipitated by 9.1C3 MAb was of 40kDa but also 9.1C3 MAb bound specifically to LAIR-1 cDNA transfected COS7 cells as well as recognized LAIR-1 fusion protein in ELISA. This finding provided the first evidence that LAIR-1 expresses on hematopoietic progenitor, implicating its role in the regulation of hematopoiesis at early stage. PMID- 14559248 TI - Degranulation and superoxide production depend on cholesterol in PLB-985 cells. AB - The effect of agents disrupting cholesterol-rich microdomains of the cell membrane was studied on the chemoattractant receptor (FPR and FRPL1) coupled effector responses of promyelocytic PLB-985 cells. Both methyl-beta-cyclodextrin (MbetaCD) and filipin III inhibited exocytosis of primary granules and O(2)(.-) production induced by stimulation of either chemotactic receptor. Alteration of calcium homeostasis of MbetaCD-treated cells does not account for the impairment of the effector responses. Disruption of microfilaments by cytochalasin B (CB) partially reverses the inhibitory effect of cholesterol depletion. Our results provide functional support for the involvement of cholesterol-rich membrane domains in the signaling of chemotactic receptors and call the attention to the possible role of microfilaments in the organization of lipid microdomains. PMID- 14559249 TI - Identification of rate-limiting steps in yeast heme biosynthesis. AB - The heme biosynthesis pathway in the yeast Saccharomyces cerevisiae is a highly regulated system, but the mechanisms accounting for this regulation remain unknown. In an attempt to identify rate-limiting steps in heme synthesis, which may constitute potential regulatory points, we constructed yeast strains overproducing two enzymes of the pathway: the porphobilinogen synthase (PBG-S) and deaminase (PBG-D). Biochemical analysis of the enzyme-overproducing strains revealed intracellular porphobilinogen and porphyrin accumulation. These results indicate that both enzymes play a rate-limiting role in yeast heme biosynthesis. PMID- 14559250 TI - Type 1 angiotensin II receptor-associated protein ARAP1 binds and recycles the receptor to the plasma membrane. AB - The carboxyl terminus of the type 1 angiotensin II receptor (AT(1)) plays an important role in receptor phosphorylation, desensitization, and internalization. The yeast two-hybrid system was employed to isolate proteins associated with the carboxyl terminal region of the AT(1A) receptor. In the present study, we report the isolation of a novel protein, ARAP1, which promotes recycling of AT(1A) to the plasma membrane in HEK-293 cells. ARAP1 cDNA encodes a 493-amino-acid protein and its mRNA is ubiquitously expressed in rat tissues. A complex of ARAP1 and AT(1A) was observed by immunoprecipitation and Western blotting in HEK-293 cells. In the presence of ARAP1, recycled AT(1A) showed a significant Ca(2+) release response to a second stimulation by Ang II 30 min after the first treatment. Immunocytochemical analysis revealed co-localization of recycled AT(1A) and ARAP1 in the plasma membrane 45 min after the initial exposure to Ang II. Taken together, these results indicate a role for ARAP1 in the recycling of the AT(1) receptor to the plasma membrane with presumable concomitant recovery of receptor signal functions. PMID- 14559251 TI - Changes in Ca2+ handling in adult MG29-deficient skeletal muscle. AB - It was reported that a lack of Mitsugumin29 (MG29), a protein expressed at the triad junction, caused morphological changes in sarcoplasmic reticulum and T tubules, reduced twitch/tetanus ratio, and increased susceptibility to fatigue in adult skeletal muscle and dysfunction of store-operated Ca2+ entry (SOC) in embryonic and neonatal muscles. To deepen our understanding of the role of MG29 in the Ca2+ handling in adult skeletal muscle,Ca2+ stores of wild-type and mutant muscle fibers were depleted by repetitive high-K+ treatments in a Ca2+-free medium. Although wild-type muscle showed only minor caffeine contracture after high-K+ response had disappeared, the mutant muscle showed remarkable caffeine contracture under the conditions used, suggesting functional compartmentalization of the Ca2+-store in the mutant. Activation of SOC in adult mutant muscle was observed upon the voltage-sensitive store depletion as is true with the wild-type muscle. Thus MG29 is not involved in the SOC activation at variance with the previous conclusion with immature muscles. PMID- 14559252 TI - Hexazinone and simazine dissipation in forestry field nurseries. AB - Hexazinone and simazine field dissipation was studied in two different soils from Spain (Toledo and Burgos), devoted to forest nurseries for Pinus nigra. Laboratory experiments (adsorption-desorption isotherms, leaching experiment and degradation study) were carried out to determine possible mechanisms of dissipation. Higher adsorption was observed for hexazinone in Toledo (KfT = 0.69) compare to in Burgos soil (KfB = 0.20) probably due to the higher organic matter (OM) content of Toledo soil. No differences in adsorption were obtained for simazine in both soils (KfT = 1.27; KfB = 1.34). In every case, adsorption was higher for simazine than for hexazinone, in both soils. The total recovery of hexazinone in the leachates from handpacked soil columns was higher in Burgos (100%) than in Toledo (80%), because of the larger adsorption of hexazinone in this last soil. No differences in simazine leaching between both soils were found, although the total amount of pesticide recovered in leachates (40% in the two soils) was lower for simazine than for hexazinone. Finally, lower degradation was found in Burgos (t1/2 = 91 d) vs Toledo (t1/2 = 47 d), directly related with the high OM content of Toledo. No half-life was calculated for simazine in Toledo because no changes in herbicide soil content were observed during the period of time studied. In the case of Burgos, the half-life for simazine was 50 days. The field residues study showed larger persistence of simazine than hexazinone mainly due to the higher adsorption and lower mobility of simazine in the two soils. The lower persistence of hexazinone in Toledo soil than in Burgos soil is related to the larger rainfall occurred in this soil besides the higher degradation of this herbicide observed in Toledo soil. The much lower temperature in Burgos than in Toledo soil during winter contribute to the higher persistence of the two herbicides in Burgos soil. PMID- 14559253 TI - The environmental occurrence of hexabromocyclododecane in Sweden. AB - The brominated flame retardant hexabromocyclododecane (HBCD) is extensively used in Europe, but data on the environmental concentrations of this chemical are scarce. A first screening has been performed concerning the environmental occurrence of HBCD in Sweden, a country where the chemical is not produced and the current industrial use is very limited. Possible emission sources were identified through a systematic analysis of the use in a life cycle perspective. In addition to a few point sources, diffuse emissions from polymeric products are possible. Measurements have been performed close to certain possible point sources, in the urban environment and in remote regions, and included air, deposition, water, soil, sediments, sludge, biota and foodstuffs. HBCD was detected in all media analysed and in all environments. The relatively high concentrations detected in herring and foodstuffs provide evidence for bioaccumulation of HBCD. The presence of HBCD in remote background air implies that HBCD has potential for long-range atmospheric transport. There are also some indications that diffuse emissions of HBCD occur in the urban environment. PMID- 14559254 TI - Development and validation of a method for analysis of "dioxin-like" PCBs in environmental samples from the steel industry. AB - A method, previously used for determination of 2,3,7,8-substituted polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs), has been modified for quantitative analysis of "dioxin-like" polychlorobiphenyls (PCBs) in environmental samples from the steel industry. The existing sample clean-up procedure, involving liquid chromatography on multi layered silica and Florisil columns, has been extended to include a third chromatography stage on a basic alumina stationary phase. The additional clean-up stage is required for PCB analysis in order to eliminate interferences from relatively large concentrations of saturated cyclic and aliphatic hydrocarbons. Samples were analysed for WHO-12 congeners using high resolution gas chromatography/high resolution mass spectrometry (HRGC/HRMS) and standard solutions of the method US EPA 1668A. Replicate analysis of method blanks revealed background contamination for PCBs 118, 105 and 77, which are generally abundant in ambient air. These contaminants were taken into account using a subtraction method. The entire procedure was validated by replicate analysis (N = 3) of a certified reference sediment. The RSD for each WHO-12 congener was below 15%, 13C12-labelled PCB internal standard recoveries were in the range 70-95%. A waste dust sample collected in the electrostatic precipitator of a UK sinter plant was analysed for determination of PCDD/Fs and WHO-12 PCBs and exhibited a PCDD/F I-TEQ of 148.5 +/- 21.2 ngkg(-1) and a WHO-TEQ of 7.2 +/- 1.5 ngkg(-1). WHO-12 congeners contributed only 4.6% to the overall TEQ and PCB 126 was the major congener contributing to the WHO-TEQ (96%). The contribution to the overall TEQ of the waste dust sample was mainly attributed to PCDF followed by PCDD, which accounted for 86.6% and 8.7% to the overall TEQ, respectively. PMID- 14559255 TI - Trends in spatial and temporal levels of persistent organic pollutants in Lake Erie sediments. AB - A Lake Erie sediment survey was conducted in 1997 to characterize spatial trends in contamination, and for comparison with a 1971 survey to assess any changes in environmental quality since the advent of measures to reduce contaminant sources. Contaminant data for some contaminant classes in 1971 was based on analysis of frozen archived samples, thereby allowing a direct comparison between surficial sediment contamination in 1971 and 1997 based on modern analytical methods. Lake wide contaminant concentrations of polychlorinated biphenyls (PCBs) and organochlorine compounds including hexachlorobenzene and the DDT compounds in 1997 were substantially lower, compared to levels in samples collected in 1971. Lake-wide average sediment PCB levels were found to have decreased roughly 70% from 136 ng/g in 1971 to 43 ng/g in 1997. Similarly, reductions in other compound classes ranged from 40% to 80%. In 1997, some individual contaminants classes including PCBs exhibited a spatial trend toward increasing sediment contamination from the eastern basin to the western basin, and from the north-central basin to the south-central basin. Levels of organic contaminants in sediments in some areas of Lake Erie still exceeded the strictest Canadian Federal and Ontario Provincial guidelines. However, exceedances of guidelines describing contaminated environments in 1997 were predominantly restricted to the western basin and near shore sites in the southern part of the central basin. PMID- 14559256 TI - Method for the determination of organophosphate insecticides in water, sediment and biota. AB - A procedure for the determination of 13 organophosphate insecticides (OPs) in water, sediment and biota at low ppb levels is described. Samples were extracted with dichloromethane or acetone/hexane and cleaned up with micro-column silica gel chromatography. Measurements were made by dual capillary column gas chromatography using both nitrogen-phosphorus (NPD) and electron capture (ECD) detection. Recoveries from fortified water samples ranged from 76% to 102% for all sample types. Practical detection limits ranged between 0.003 and 0.029 microg/l in natural water samples, 0.0004-0.005 microg/g w.w. for sediments, and 0.001-0.005 microg/g w.w for biota using the NPD and ECD method. Losses in sediments were experienced when sulphur was removed. Precision and accuracy were not affected in sediment samples where sulphur was not removed. PMID- 14559257 TI - Formation of chlorinated and brominated dioxins and other organohalogen compounds at the pilot incineration plant VERONA. AB - Experiments were conducted at the VERONA pilot plant, an incineration plant with stationary grate and separate post-combustion chamber, using wood and propane as basic combustible materials and with controlled dosage of various bromine-, chlorine- and copper-containing compounds. The behaviour of the following compounds was studied in the combustion chamber, after the post-combustion chamber and after the heat exchanger: polychlorinated phenols (PCPh), polybrominated phenols (PBrPh), polychlorinated benzenes (PCBz), polybrominated benzenes (PBrBz), polychlorinated dioxins and furans (PCDD/F) and polybrominated dioxins and furans (PBDD/F). The bromine co-incineration leaded to very high bromophenol concentrations after the post-combustion chamber. The formation of brominated and mixed-halogenated phenols and the further reaction to halogenated dioxins is apparently a relevant reaction mechanism for dioxin formation in processes involving bromine. This assumption is supported by the high formation rates of PBDD/F found in the heat exchanger, which were 4-20 times higher than those of PCDD/F. Moreover, the strong correlations found between the formation rates of PCPh, PCBz and PCDD/F in the heat exchanger indicate that in addition considerable new formation of dioxins takes place through de novo synthesis. Experiments involving the variation of primary operational parameters and fuel properties have shown that the quality of post-combustion plays a much greater role than the other parameters. Furthermore, it became apparent that the congeners of the chlorophenols and of the chlorobenzenes, measured in various incineration stages, do not correlate closely with the dioxin concentrations after the heat exchanger. PMID- 14559258 TI - Uptake and depuration of toxic halogenated aromatic hydrocarbons by the American oyster (Crassostrea virginica): a field study. AB - Uptake and depuration of toxic chlorinated compounds such as planar polychlorinated biphenyls (PCBs 77, 126, 169), 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD) and 2,3,7,8-tetrachlorodibenzofuran (TCDF) were monitored during a 50-day field study where American oysters (Crassostrea virginica) were transplanted back and forth between a heavily polluted area (Houston Ship Channel, SC) and a relatively unimpacted area (Hanna's Reef, HR) within Galveston Bay, TX. In general, low molecular weight, less lipophilic compounds accumulated in the oysters to a larger extent than high molecular weight ones. Estimated half lives for planar PCB congeners 77 and 126 were 28 and 51 days, respectively for depuration of newly contaminated oysters (HR-SC-HR) while longer half-lives (42 and 60 days, respectively) were observed for the same compounds as they were eliminated from chronically contaminated individuals (SC-HR). Estimated half lives for 2,3,7,8-TCDD and 2,3,7,8-TCDF were 35 and 36 days, respectively and were similar to the tetrachlorinated biphenyls (PCBs 77 and 81). Compared with ortho-substituted PCB congeners of the same chlorination level, the more toxic PCBs take longer to depurate from the oysters. With few exceptions, elimination of all toxic compounds investigated proceeded at a slower rate from the chronically exposed population that from the newly contaminated one. PMID- 14559259 TI - Wet peroxide degradation of atrazine. AB - The high temperature (150-200 degrees C), high pressure (3.0-6.0 MPa) degradation of atrazine in aqueous solution has been studied. Under these extreme conditions atrazine steadily hydrolyses in the absence of oxidising agents. Additionally, oxygen partial pressure has been shown not to affect atrazine degradation rates. In no case mineralisation of the parent compound was observed. The addition of the free radical generator hydrogen peroxide to the reaction media significantly enhanced the depletion rate of atrazine. Moreover, partial mineralisation of the organics was observed when hydrogen peroxide was used. Again, oxygen presence did not influence the efficiency of the promoted reaction. Consecutive injections of hydrogen peroxide throughout the reaction period brought the total carbon content conversion to a maximum of 65-70% after 40 min of treatment (suggesting the total conversion of atrazine to cyanuric acid). Toxicity of the effluent measured in a luminometer decreased from 93% up to 23% of inhibition percentage. The process has been simulated by means of a semi-empirical model. PMID- 14559260 TI - Concentrations of dioxin-like PCB congeners in unweathered Aroclors by HRGC/HRMS using EPA Method 1668A. AB - We have determined the congener compositions of nine commercial Aroclor products of polychlorinated biphenyls (PCBs) to the sub-part-per-million level using high resolution gas chromatography combined with high-resolution mass spectrometry according to US Environmental Protection Agency (EPA) Method 1668A. These Aroclor composition data should allow improved characterization and risk assessment of PCB contamination at hazardous waste sites, particularly for dioxin-like PCB congeners. By combining the data on the concentrations of each dioxin-like congener with its World Health Organization toxicity equivalency factor, we have established dioxin toxic equivalent concentrations for each pure Aroclor product. PMID- 14559261 TI - Remediation of soil contaminated with dioxins by subcritical water extraction. AB - The effectiveness of subcritical water extraction (SCWE) was examined for removing dioxins from contaminated soil. Most dioxins in the soil sample were reduced at 300 degrees C or more, but decreased dioxin concentrations were also observed at 150 degrees C. After 4 h of extraction, 99.4%, 94.5% and 60% of PCDDs were removed from samples at 350, 300 and 150 degrees C, respectively. It was also determined that degradation of dioxins had occurred, since the sum of dioxins in the soil plus water extracts after the experiments had considerably decreased. This study revealed that pressurizing is not essential for the removal of dioxins. Reduction was complete within 30 min at 350 degrees C; however, it took a much longer time at lower temperatures. The results of addition experiments in which OCDDs were added to different types of soil samples have shown that dechlorination is one of the major reaction pathways. After addition of OCDD to soil samples, experiments were carried out to examine in detail the degradation pathways of PCDDs. The removal rates and congener profiles varied among soil types. Although it was previously assumed that removal rates and congener profiles depended on the chemical components in soil, nonparametric statistical analysis revealed no significant relationship between the rate of reduction and elements present in the soil. It was confirmed from isomer patterns that dechlorination of the 2,3,7,8-positions in PCDDs takes place somewhat faster than for the 1,4,6,9-positions. PMID- 14559262 TI - De novo formation characteristics of dioxins in the dry zone of an iron ore sintering bed. AB - The objectives of this work are to understand the details of the mechanism of dioxin formation in the part of a sintering bed termed the dry zone, and to obtain ideas on how to prevent their formation. Sinter mixtures of various composition types were heated in a packed bed reactor, and dioxins in the outlet gas and in the sinter mixture residue were measured. The dioxin formation potential of a simple sinter mixture composed of iron ore, coke and limestone was markedly lower than that of fly ash from a municipal solid waste incinerator (MSWI). In consideration of this result, a series of experiments were conducted using a sinter mixture impregnated with CuCl2. Experimental results showed that dioxin formation was temperature-dependent in the range of 300-550 degrees C, with the maximum observed at around 300 degrees C, which was quite similar to that of fly ash from the MSWI. The homologue distribution of PCDD/Fs in gas and solid reflected the possible difference in carbonaceous materials in coke and activated coke. Gaseous hydrogen chloride acted as a chlorinating reagent for dioxin formation. PMID- 14559263 TI - Influence of coexisting surface-active agents on leachability of dioxins in raw and treated fly ash from an MSW incinerator. AB - The leaching behavior of dioxins from raw and treated fly ash (FA) under the coexistence of several types of surface-active agents (SAAs) was examined by batch leaching tests to obtain significant information not only for evaluating leachability of dioxins as hydrophobic organic pollutants (HOPs) under the severe environment in which SAAs coexist, but also for evaluating the reduction efficiency of dioxin-leachability from the treated FA. Enhancement of dioxin leachability by sufficient addition of SAAs was observed in the leaching test, whereas addition of a smaller quantity depressed the leachability. The higher the degree of chlorination in PCDDs/PCDFs homologues, the more effectively the SAAs enhanced the leachability. Moreover, there was a large difference in the potential for leachability enhancement due to each surface activity, based on critical micelle concentration (CMC) of the SAAs although Aldrich humic acid enhanced the leachability at a much lower concentration than CMC. A comparison of the leachability in FA treated by several methods showed a difference in the reduction efficiency of leachability, depending on the adsorption of SAAs by each treated FA. PMID- 14559264 TI - Photochemical transformations of tetrabromobisphenol A and related phenols in water. AB - A method was developed for studies of the phototransformation at UV irradiation of aqueous solutions of tetrabromobisphenol A (TBBPA), tribromobisphenol A (TriBBPA), tetrachlorobisphenol A (TCBPA), 2,4-dichlorophenol at various pHs as well as 2-chlorophenol, 2-bromophenol, 3,4-dichlorophenol and bisphenol A at pH 11. The absorbance spectra of the compounds and the emission spectra of the light source were determined and used to calculate disappearance quantum yields of the photochemical reactions that were taking place. No major differences between the disappearance quantum yields of TBBPA and TCBPA were observed at pH 10, while the disappearance quantum yield of TriBBPA was approximately two times higher. The rate of decomposition of TBBPA was six times higher at pH 8 than at pH 6. Identification of the degradation products of TBBPA and TriBBPA, by GC-MS analysis and by comparison to synthesised reference compounds, indicated that TBBPA and TriBBPA decompose via different mechanisms. Three isopropylphenol derivatives; 4-isopropyl-2,6-dibromophenol, 4-isopropylene-2,6-dibromophenol and 4-(2-hydroxyisopropyl)-2,6-dibromophenol, were identified as major degradation products of TBBPA while the major degradation product of TriBBPA was tentatively identified as 2-(2,4-cyclopentadienyl)-2-(3,5-dibromo-4-hydroxyphenyl)propane. PMID- 14559265 TI - Levels of PCDDs, PCDFs and Co-PCBs in human milk in Saitama, Japan, and epidemiological research. AB - From 1998 to 2000, the dioxin levels were measured in the milk of 299 mothers who lived in Saitama prefecture, Japan. Factors that influenced the dioxin levels were investigated based on a questionnaire given to the milk donors at that time. It was found that the dioxin levels in the milk of the mothers who smoked were lower than those of non-smokers, and the dioxin levels were generally low in the mothers who were heavy smokers. The average dioxin levels in the milk of mothers who had been breast-fed were higher than those given formula, and there was a significant difference in the dioxin congeners. It was also found that the dioxin levels in milk of the women who regularly consumed fish and shellfish were generally higher. There was a strong correlation between summation operator TEQ and PCB126. The data suggested that PCB126 could be a useful indicator for the simplified analysis of dioxin in human milk. PMID- 14559266 TI - Levels of PCDDs, PCDFs and PCBs in Belgian and international fast food samples. AB - Congener-specific analyses of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and polychlorinated biphenyls (PCBs) were performed on twenty-eight non-pooled fast food samples collected in Belgium, Switzerland, Czech Republic, United States of America and Australia. PCDD/F and PCB concentrations for the four investigated types of meals were very low. PCDD/F values ranged from non-detected to 1.40 pg WHO-TEQ/g fat and from 0.79 to 2.08 pg WHO-TEQ/g fat for lower and upper bound, respectively. Major contributors to the PCDD/F TEQ were 1,2,3,4,7,8-HxCDD, 1,2,3,6,7,8-HxCDD, 2,3,7,8-TCDF and 2,3,4,7,8 PeCDF. The relative contribution of PCBs to the total TEQ was 68%. For adults, an average estimated intake was 6.7 pg WHO-TEQ/kg bw/month, including consumption of all types of analyzed meals, representing 9.5% of the PTMI. For child, a value of 14.5 pg WHO-TEQ/kg bw/month was obtained, representing 20.6% of the PTMI. PMID- 14559267 TI - A new model of tropospheric hydroxyl radical concentrations. AB - A chemistry model of the global troposphere is presented which focuses on the hydroxyl radical, OH. Global distributions of OH are calculated based on known chemical reaction pathways, experimentally measured values of precursor species O3, H2O, NOx (defined as NO+NO2), CO, CH4, and actinic flux (which includes the effects of cloud cover and O3 column absorption). Model grid resolution is 1 km in altitude by 10 degrees latitude, and zonally divided into land or ocean. Species are calculated as seasonal averages. Global annual mean OH in the troposphere (up to 14 km altitude) is calculated to be 9.2 x 10(5) molcm(-3) with averages of 9.8 x 10(5) in the northern hemisphere, and 8.5 x 10(5) in the southern hemisphere. Global CO and CH(4) oxidation rates by OH are calculated to be 1840 Tgyear(-1) and 580 Tgyear(-1), respectively. OH is found to be most sensitive to O3 and H2O concentrations, as well as to the photolysis rate of O3 to O1D. Sensitivity of CO and CH4 oxidation rates to cloud presence shows an inverse relationship to cloud amount and optical depth. Model results are shown to be consistent with results from two other published models. PMID- 14559268 TI - Characterization of aerosols from biomass burning--a case study from Mizoram (Northeast), India. AB - Physical and optical properties of biomass burning aerosols in Northeastern region, India analyzed based on measurements made during February 2002. Large spatial extent of Northeastern Region moist tropical to moist sub-tropical forests in India have high frequency of burning in annual dry seasons. Characterization of resultant trace gases and aerosols from biomass burning is important for the atmospheric radiative process. Aerosol optical depth (AOD) observed to be high during burning period compared to pre- and post-burning days. Peak period of biomass burning is highly correlated with measured AOD and total columnar water vapor. Size distribution of aerosols showed bimodal size distribution during burning day and unimodal size distribution during pre- and post-burning days. Size distribution retrievals from biomass burning aerosols show dominance of accumulation mode particles. Weighted mean radius is high (0.22 microm) during burning period. Columnar content of aerosols observed to be high during burning period in addition to the drastic reduction of visibility. During the burning day Anderson sampler measurements showed dominance of accumulation mode particles. The diurnal averaged values of surface shortwave aerosol radiative forcing af biomass burning aerosols varies from -59 to -87 Wm(-2) on different days. Measured and modeled solar irradiances are also discussed in the paper. PMID- 14559269 TI - Application of the SO4(2-)/Se tracer technique to study SO2 oxidation in cloud and fog on a time scale of minutes. AB - We have demonstrated the use of Se as a tracer to quantitatively determine in situ SO4(2-) production from SO2 oxidation in clouds and fogs. Until now, it has not been possible to study the kinetics of SO2 oxidation because the aerosol sampling interval for Se determination was limited to 2 h or longer. Here we report results of 5-min aerosol measurements carried out at Lahore, Pakistan, during January 9-11, 2001, using new methodology for Se analysis coupled with hydride generation and ICP-MS detection. These improvements will enable the tracer technique to determine in situ SO4(2-) production in clouds and fogs on a time scale of several minutes and possibly 1 min. The method may prove useful for kinetic studies of in-cloud SO2 oxidation and in the study of other phenomena such as atmospheric mixing, cloud drop lifetimes, and aerosol formation that occur on the time scale of a few minutes. PMID- 14559270 TI - Source profiles for industrial, mobile, and area sources in the Big Bend Regional Aerosol Visibility and Observational study. AB - Representative PM2.5 and PM10 source emissions were sampled in Texas during the Big Bend Regional Aerosol Visibility and Observa (BRAVO) study. Chemical source profiles for elements, ions, and carbon fractions of 145 samples are reported for paved and unpaved road dust, soil dust, motor vehicle exhaust, vegetative burning, four coal-fired power stations, an oil refinery catalytic cracker, two cement kilns, and residential meat cooking. Several samples were taken from each emitter and source type, and these were averaged by source type, and in source subgroups based on commonality of chemical composition. The standard deviation represents the variability of the chemical mass fractions. BRAVO profiles differed in some respects from profiles measured elsewhere. High calcium abundances in geological dust, high selenium abundances in coal-fired power stations, and high antimony abundances in oil refinery catalytic cracker emissions were found. Abundances of eight thermally evolved carbon fractions [Atmos. Environ. 28 (15) (1994) 2493] differ among combustion sources, and a Monte Carlo simulation demonstrates that these differences are sufficient to differentiate among several carbon-emitters. PMID- 14559271 TI - Influence of plants on the methane emission from sediments. AB - The previous theory [Chem. Global Change Sci. 3 (2001) 33; Chemosphere 50 (2003) 191] of methane emission is applied to vegetated sediments. The presence of roots in a sediment is taken into account. It is assumed that methane and nitrogen enter a sediment through channels existing in plants and roots. The rate of methane and nitrogen transport through plants and roots is proportional to the difference in concentrations in the layer and on the upper surface. It is established that as the vegetation density increases, the rate of methane transport increases so that with sufficient vegetation density, almost all methane passes to the atmosphere through plants. In this case, the value of bubble emission decreases to zero. The nitrogen transport rate through plants first increases and then decreases with increasing the vegetation density. The theory qualitatively and quantitatively describes the dependence of methane concentration on depth in the presence of plants. A comparison with the available experimental data on dissolved methane concentration and bubble composition indicates satisfactory agreement. PMID- 14559272 TI - Changes in the grassland-forest boundary at Ta-Ta-Chia long term ecological research (LTER) site detected by stable isotope ratios of soil organic matter. AB - The carbon isotope analysis [delta13C values] of organic samples can be a useful research in ecological studies because delta13C values are indicative of the plant source. This study investigated the changes in plant communities along the grassland-forest boundary in the alpine forest at Ta-Ta-Chia long term ecological research (LTER) site in central Taiwan using carbon isotope data. The aim of this study was focused on the forest fire affected the change of vegetation community. Four pedons from grassland dominated by Miscanthus transmorrisonensis (pedons 1 and 2), transition zone by Tsuga and Yushania nittakeyamensis (pedon 3), and forest zone by Tsuga and nittakeyamensis (pedon 4) were examined. Soil organic matter (SOM) delta13C values in the upper soil horizon were similar to delta13C values of the overlaying vegetation types. This indicates that the boundary between these plant communities remained the same in the past decades. The delta13C values of the grassland SOM ranged from -19.4 per thousand to -24.1 per thousand, showing decrease with soil depth. This suggests that C4 plants (transmorrisonensis) have replaced C3 plants of Tsuga and nittakeyamensis. The delta13C values of the Tsuga forest area (pedon 4) range from -27.0 per thousand to -23.5 per thousand and showed only slight change with soil depth, implying that C3 plants have remained the major species in the forest. PMID- 14559273 TI - Guaiane dimers from Xylopia vielana. AB - From the leaves of Xylopia vielana (Annonaceae) two dimeric guaianes named vielanins D and E were isolated and structurally elucidated by mass and NMR spectroscopy. Vielanin D and E consist of bridged ring systems formally representing the Diels-Alder products from the hypothetical guaiane-type monomers. Due to a hemiketal function at C-8' both compounds occurred as epimeric mixtures. PMID- 14559274 TI - Ring A-seco mosquito larvicidal limonoids from Turraea wakefieldii. AB - Five novel limonoids, 1-5, were isolated from the root bark of Turraea wakefieldii and were characterized as tecleaninoid derivatives. This is the first report of the natural occurrence of tecleanin-type limonoids with a five-membered ring A-seco structure for which we propose the name neotecleanins. The relative stereochemical structures of compounds 1-5 were established on the basis of NMR spectroscopy. The absolute stereochemical structure of 5 was confirmed by X-ray diffraction methods. In mosquito larvicidal assays, compounds 1, 2 and 4 showed dose-dependent larvicidal activity against larvae of Anopheles gambiae s.s. PMID- 14559275 TI - Detection of unusual carotenoid esters in fresh mango (Mangifera indica L. cv. 'Kent'). AB - The carotenoid pattern of mango cv. 'Kent' was investigated by LC-(APcI)MS analyses. In solvent extracts from the mesocarp an unusual carotenoid ester was identified as violaxanthin dibutyrate. For unequivocal identification of butyric acid by an independent method, total lipids were isolated by solvent extraction from the fruit flesh and analyzed by GC after saponification and subsequent methylation. Thus, evidence of butyric acid (1.6 area%) was provided. To the best of our knowledge, this is the first report on a xanthophyll dibutyrate in plants. Additionally, further carotenoid peaks were tentatively assigned to 9-cis violaxanthin and neochrom or luteoxanthin, respectively, by their UV/vis and MS data of the saponified extracts. PMID- 14559276 TI - Artoindonesianins X and Y, two isoprenylated 2-arylbenzofurans, from Artocarpus fretessi (Moraceae). AB - Two isoprenylated 2-arylbenzofurans, artoindonesianins X and Y (1-2), together with seven known flavonoids, have been isolated from the roots and tree bark of Artocarpus fretessi. Their structures were established on the basis of spectral analysis. Compounds 1 and 2 showed moderate activity against the brine shrimp Artemia salina. PMID- 14559277 TI - Seco-labdane type diterpenes from Excoecaria agallocha. AB - Labdane-type diterpenes, called excoecarins S, T1, and T2 were isolated from the resinous wood of Excoecaria agallocha, along with three known compounds, ent-12 oxo-2,3-secobeyer-15-ene-2,3-dioic acid, agallochin H, and ent-15-epoxy-beyerane 3alpha-ol. Their structures were elucidated on the basis of spectroscopic data, chemical evidence, and X-ray analysis. PMID- 14559278 TI - Pseudoguaianolides from the flowers of Parthenium hysterophorus. AB - Chemical investigation on the flowers of Parthenium hysterophorus has resulted in the isolation of four new pseudoguaianolides, hysterones A-D along with the known compounds, parthenin, coronopilin, 2beta-hydroxycoronopilin and tetraneurin-A. The structures of the new compounds were established by interpretation of their spectral (1D and 2D NMR) data. The X-ray crystallographic analysis of hysterones A and C was also carried out. PMID- 14559280 TI - p-coumaric acid esters from Tanacetum longifolium. AB - Two new long chain alkyl p-coumaric acid esters (2-3) along with eicosanyl trans p-coumarate (1) were isolated from chloroform extract of the roots of Tanacetum longifolium. The structures of new compounds were assigned as 21' hydroxyheneicosanyl-4-hydroxy-(cis and trans) p-coumarate (2a, 2b) and 27' hydroxy heptacosanyl-cis-p-coumarate (3) by extensive chromatographic and spectroscopic analysis and by comparison with literature data of known compounds. PMID- 14559279 TI - Pentacyclic triterpenoid and saponins from Gambeya boukokoensis. AB - Chemical studies of the EtOAc extract of Gambeya boukokoensis Aubr. et Pellegr. stem bark led to the isolation of eight compounds. Three of them were elucidated as new compounds and designated as: gamboukokoensein A, 1alpha,2alpha,3beta,19alpha,23-pentahydroxyurs-12-en-28-oic acid; gamboukokoenside A, 2beta,3beta,6beta,28-tetrahydroxyolean-12-en-23-oic acid 23-O alpha-L-arabinopyranosyl ester and gamboukokoenside B, 6beta,28-dihydroxy-3 oxoolean-12-en-23-oic acid 23-O-alpha-L-arabinopyranosyl ester. The other five compounds were known and identified as myrianthic acid, protobassic acid, oleanolic acid, erythrodiol and chondrillasterol. Their structures were elucidated on the basis of one and two dimensional NMR spectroscopic techniques, FABMS, ESMS and chemical evidence. PMID- 14559281 TI - Benzoquinone derivatives of Myrsine africana and Maesa lanceolata. AB - The fruits of Myrsine africana afforded two new benzoquinone derivatives, methylvilangin and methylanhydrovilangin. On the other hand, from the fruits of Maesa lanceolata two more novel compounds; 2,5-dihydroxy-3-(nonadec-14-enyl) benzoquinone and lanciaquinone were isolated. Their structural elucidation was achieved by spectroscopic measurements including 2D NMR experiments. PMID- 14559282 TI - Anthrasesamones from roots of Sesamum indicum. AB - Three anthraquinones, named anthrasesamones A, B and C, were isolated from the roots of Sesamum indicum, and their respective structures were determined to be 1 hydroxy-2-(4-methylpent-3-enyl)anthraquinone, 1,4-dihydroxy-2-(4-methylpent-3 enyl)anthraquinone and 2-chloro-1,4-dihydroxy-3-(4-methylpent-3 enyl)anthraquinone on the basis of spectroscopic evidence. Two known anthraquinones were also isolated for the first time from S. indicum roots and characterized as 2-(4-methylpent-3-enyl)anthraquinone and (E)-2-(4-methylpenta 1,3-dienyl)anthraquinone. Anthrasesamone C is a rare chlorinated anthraquinone in higher plants. PMID- 14559283 TI - Anthocyanins acylated with gallic acid from chenille plant, Acalypha hispida. AB - Three anthocyanins were isolated from the red flowers of chenille plant, Acalypha hispida Burm. (Euphorbiaceae) by a combination of chromatographic techniques. Their structures were elucidated mainly by homo- and heteronuclear nuclear magnetic resonance spectroscopy and electrospray mass spectrometry, and supported with complete assignments of 13C NMR resonances. The novel pigment, cyanidin 3-O (2"-galloyl-6"-O-alpha-rhamnopyranosyl-beta-galactopyranoside) (5%), contains the disaccharide robinoside. The other anthocyanins were identified as cyanidin 3-O (2"-galloyl-beta-galactopyranoside) (85%), and cyanidin 3-O-beta galactopyranoside (5%). Anthocyanins acylated with gallic acid have previously been identified in species from the families Nymphaeaceae and Aceraceae, and tentatively in Abrus precatorius (Leguminosae). PMID- 14559284 TI - Coumarins from Malaysian Micromelum minutum. AB - In a continuation of our study of the Rutaceae, detailed chemical investigation on Micromelum minutum (Rutaceae) collected from Sepilok, Sabah, Malaysia gave four new coumarins. The structures of the coumarins have been fully characterised by spectroscopic methods as 3",4"-dihydrocapnolactone 1, 2',3' epoxyisocapnolactone 2, 8-hydroxyisocapnolactone-2',3'-diol 3 and 8-hydroxy-3",4" dihydrocapnolactone-2',3'-diol 4. PMID- 14559285 TI - A flavanone and a dihydrodibenzoxepin from Bauhinia variegata. AB - Phytochemical analysis of the root bark of Bauhinia variegata Linn yielded a new flavanone, (2S)-5,7-dimethoxy-3',4'-methylenedioxyflavanone (1) and a new dihydrodibenzoxepin, 5,6-dihydro-1,7-dihydroxy-3,4-dimethoxy-2-methyldibenz [b,f]oxepin (2) together with three known flavonoids (3-5). The structures of the new compounds were determined on the basis of spectral studies. PMID- 14559286 TI - Polyoxygenated flavonoids from Eugenia edulis. AB - Leaves of Eugenia edulis contain the new polyoxygenated flavonoid derivatives, gossypetin-3,8-dimethyl ether-5-O-beta-glucoside; gossypetin-3,5-dimethyl ether, and myricetin-3,5,3'-trimethyl ether. In addition, ten known polyphenolics were also isolated and identified. All structures were established on the basis of chemical and spectral evidence, including ESI-MS and 13C NMR. PMID- 14559287 TI - Flavonol glycosides of Warburgia ugandensis leaves. AB - Four new flavonol gycosides: kaempferide 3-O-beta-xylosyl (1-->2)-beta-glucoside, kaempferol 3-O-alpha-rhamnoside-7,4'-di-O-beta-galactoside, kaempferol 3,7,4'-tri O-beta-glucoside and quercetin 3-O-[alpha-rhamnosyl (1-->6)] [beta-glucosyl (1- >2)]-beta-glucoside-7-O-alpha-rhamnoside, were characterized from a methanolic leaf extract of Warburgia ugandensis. The known flavonols: kaempferol, kaempferol 3-rhamnoside, kaempferol 3-rutinoside, myricetin, quercetin 3-rhamnoside, kaempferol 3-arabinoside, quercetin 3-glucoside, quercetin, kaempferol 3 rhamnoside-4'-galactoside, myricetin 3-galactoside and kaempferol 3-glucoside were also isolated. Structures were established by spectroscopic and chemical methods and by comparison with authentic samples. PMID- 14559288 TI - Alkaloids and a pimarane diterpenoid from Strychnos vanprukii. AB - From the stem of Strychnos vanprukii, a gluco-indole alkaloid, 3,4 dehydropalicoside, and a pimarane diterpenoid, 7 beta-hydroxypimara-8,15-dien-14 one, were isolated together with four known alkaloids: palicoside, 3,4,5,6 tetradehydropalicoside, akagerine and 17-O-methylakagerine. The structures of these compounds were elucidated based on spectroscopic evidence. PMID- 14559289 TI - Pinelloside, an antimicrobial cerebroside from Pinellia ternata. AB - An antimicrobial cerebroside, pinelloside, was isolated from the air-dried tubers of Pinellia ternata (Thunb.) Breit. Its structure was determined as 1-O-beta-D glucopyranosyl-(2S,3R,4E,11E)-2-(2'R-hydroxyhexadecenoylamino)-4,11-octadecadiene 1,3-diol by chemical transformation and extensive spectroscopic analyses (IR, MS, 1H and 13C NMR, DEPT as well as 2D NMR techniques HMBC, HMQC, 1H-1H COSY and NOESY). The antimicrobial assay showed that this compound was inhibitory to the growth of Bacillus subtilis, Staphylococcus aureus, Aspergillus niger and Candida albicans, with minimum inhibitory concentrations (MICs) of 20, 50, 30 and 10 microg/ml, respectively. The MICs of penicillin G against bacteria B. subtilis, S. aureus, E. coli, P. fluorescens and H. pylori were 0.80, 0.34, 0.56, 1.34 and 0.92, and those of ketoconazole against fungi A. niger, C. albicans and T. rubrum 0.90, 0.65 and 1.0 microg/ml, respectively. PMID- 14559291 TI - Pilocarpine-induced seizures in adult rats: monoamine content and muscarinic and dopaminergic receptor changes in the striatum. AB - High doses of the muscarinic cholinergic agonist pilocarpine are a useful model for investigation of the essential mechanisms for seizure generation and spread in rodents. Pilocarpine (400 mg/kg; subcutaneously) was administered in 2-month old female rats, and the content of striatum monoamines and (M(1)+M(2)) muscarinic and D(2) dopaminergic receptors was measured in the acute period. All treated animals showed peripheral cholinergic signs, stereotyped and clonic movements, tremors, seizures and the percentage mortality was approximately 63%. High performance liquid chromatography determinations, performed 24 h later, showed a decrease of striatal levels of dopamine, dihydroxyphenylacetic acid, 4 hydroxy-3-methoxy-phenylacetic acid and 5-hydroxytryptamine. Pilocarpine treatment induced downregulation of (M(1)+M(2)) muscarinic receptors and reduced the dissociation constants of (M(1)+M(2)) muscarinic and D(2) dopaminergic receptors, suggesting that these systems exert opposite effects on the regulation of convulsive activity. These and other important neurochemical changes found in the course of establishment of an epileptic focus can be observed after status epilepticus induced by pilocarpine. PMID- 14559292 TI - Antioxidant activity of compounds from the medicinal herb Aster tataricus. AB - A number of compounds were isolated from the medicinal plant Aster tataricus including shionone, epifriedelinol, quercetin, kaempferol, scopoletin, emodin, aurantiamide acetate and 1,7-dihydroxy-6-methyl-anthraquinone. The compounds were compared with regard to their ability in inhibiting hemolysis of rat erythrocytes induced by 2'-2' azobis (2-amidinopropane) dihydrochloride, lipid peroxidation using the FeSO(4)-ascorbic acid system, and generation of superoxide radicals using a phenazine methosulfate-nicotinamide adenine dinucleotide system. The effects on the Fe-bleomycin-induced DNA damage reflected pro-oxidant activity. Quercetin and kaempferol were most potent in inhibiting hemolysis, lipid peroxidation and superoxide radical generation. Scopoletin and emodin were similar to quercetin and kaempferol in inhibiting superoxide radical generation and second to them in inhibiting lipid peroxidation. Aurantiamide acetate exhibited some inhibitory activity toward superoxide radical generation. 1,7 dihydroxy-6-methyl-anthraquinone exerted an inhibitory activity only on superoxide radical generation. Shionone and epifriedelinol did not display any antioxidant activity. Quercetin and kaempferol, but not the remaining compounds, exhibited some pro-oxidant activity. PMID- 14559293 TI - Effects of municipal effluents on serotonin and dopamine levels in the freshwater mussel Elliptio complanata. AB - Sex differentiation and gametogenesis represent critical steps in the reproductive process and are subject to hormonal control by serotonin, dopamine and steroids such as estradiol-17beta and testosterone. The purpose of this study sought to examine the endocrine-disrupting activity that a primary-treated municipal effluent might have on the metabolism of biogenic amine levels. First, serotonin receptors transfected in Chinese hamster ovary (CHO) cells were used to screen for the presence of serotonin receptor agonist or antagonist. Second, one group of Elliptio complanata mussels were exposed to single compounds likely to be found in municipal wastewaters and another group was exposed in situ to the municipal effluent plume for 90 days in experimental cages. Results showed that solid phase C-8 extracts of surface water downstream a municipal effluent could activate the transport of serotonin by receptors at a distance of at least 5 km from its outfall thereby indicating the presence of serotonin mimics in the effluent dispersion plume. Levels of serotonin and monoamine oxidase (MAO) activity in nerve ganglia of mussels exposed for 90 days to the municipal effluent were, respectively, reduced and increased at a distance 10-km downstream. Injections of estradiol-17beta and nonylphenol in mussels decreased the levels of serotonin and dopamine, but increased MAO activity in the gonad and nerve ganglia. Exposure to estrogenic chemicals present in municipal effluents may therefore alter the normal metabolism of serotonin and dopamine, both of which are involved in sexual differentiation in bivalves and fish. Chemicals acting through E2 receptor-mediated pathways and serotonin receptors are likely to cause the observed effects. PMID- 14559294 TI - The heart of Daphnia magna: effects of four cardioactive drugs. AB - We used Daphnia magna bioassays to determine the LC(50) and the effects on the heart of the cardioactive drugs ouabain, verapamil, metaproterenol and metoprolol. Distinctions were made between the pharmacological and toxicological effects of these drugs and the adequacy of physicochemical characteristics of its habitat (reconstituted water). Video microscopy and digital image processing were used to study the pharmacological effects on the heart. D. magna exhibited the expected sensitivity to the reference toxicant sodium dodecyl sulfate with a LC(50) of 15.6+/-4.5 mg/l. All drugs were toxic with 48 h-LC(50) of 2.03 mg/l ouabain, 7.04 mg/l verapamil, 32.45 mg/l metaproterenol and 76.21 mg/l metoprolol. Ouabain was the most toxic and caused a positive concentration dependent inotropic effect. Verapamil caused positive chronotropic and inotropic effects, while metaproterenol showed positive concentration-dependent chronotropic effects at high concentrations (10(-3) and 10(-4) M). Metoprolol induced a positive chronotropic effect at low concentrations (10(-8), 10(-7), 10( 6) M) and a negative chronotropic effect at high concentration (10(-4) M). Ouabain, metaproterenol and metoprolol in D. magna caused similar effects to those produced in mammals. In contrast, verapamil caused opposite effects. The results suggest the presence of Na(+), K(+)-ATPase receptors to verapamil and of non-specific adrenergic receptors in heart of D. magna. PMID- 14559295 TI - Cytochrome c oxidase inhibition in the rice weevil Sitophilus oryzae (L.) by formate, the toxic metabolite of volatile alkyl formates. AB - Volatile alkyl formates are potential replacements for the ozone-depleting fumigant, methyl bromide, as postharvest insecticides and here we have investigated their mode of insecticidal action. Firstly, a range of alkyl esters, ethanol and formic acid were tested in mortality bioassays with adults of the rice weevil, Sitophilus oryzae (L.) and the grain borer, Rhyzopertha dominica (F.) to determine whether the intact ester or one of its components was the toxic moiety. Volatile alkyl formates and formic acid caused similar levels of mortality (LC(50) 131-165 micromol l(-1)) to S. oryzae and were more potent than non-formate containing alkyl esters and ethanol (LC(50)>275 micromol l(-1)). The order of potency was the same in R. dominica. Ethyl formate was rapidly metabolised in vitro to formic acid when incubated with insect homogenates, presumably through the action of esterases. S. oryzae and R. dominica fumigated with a lethal dose of ethyl formate had eight and 17-fold higher concentrations of formic acid, respectively, in their bodies than untreated controls. When tested against isolated mitochondria from S. oryzae, alkyl esters, alcohols, acetate and propionate salts were not inhibitory towards cytochrome c oxidase (EC 1.9.3.1), but sodium cyanide and sodium formate were inhibitory with IC(50) values of 0.0015 mM and 59 mM, respectively. Volatile formate esters were more toxic than other alkyl esters, and this was found to be due, at least in part, to their hydrolysis to formic acid and its inhibition of cytochrome c oxidase. PMID- 14559296 TI - Feminization of wild carp, Cyprinus carpio, in a polluted environment: plasma steroid hormones, gonadal morphology and xenobiotic metabolizing system. AB - Wild carp, Cyprinus carpio, were sampled in January and March 2000 in a section of the Anoia River (NE Spain) known to be polluted by estrogenic compounds. At each sampling time, three groups were distinguished: (1) apparently normal males; (2) apparently normal females; and (3) affected fish. The latter were characterized by the simultaneous development of male and female tissue in their gonads at a macroscopical level (six out of 31 fish sampled at this particular point), or testicular atrophy (three out of 31). Plasmatic and hepatic vitellogenin (VTG) levels and plasma testosterone (T) and estradiol (E2) were measured to observe the particular estrogenic response of the affected fish. Moreover, the response in the xenobiotic metabolizing capacity in liver was tested. This involved the analysis of mixed function oxygenase (MFO) system such as: total cytochrome P450 content, NAD(P)H cytochrome c reductases and the associated CYP1A1, EROD activity. Also, glutathione S-transferase (GST) and UDP glucuronosyltransferase (UDPGT) as detoxifying enzymes were measured. Our results showed: (1) a highly variable VTG content in all fish groups; (2) an increase in sex hormones content in March for the female group; and (3) an enhanced xenobiotics metabolism in the affected fish group, measured as total cytochrome P450, EROD activity in the January survey and cytosolic GST in March. The observed increase in VTG, sex hormones and in most of the enzymatic activities from January to March that could also be attributed to higher water temperature. PMID- 14559297 TI - Metabolic fate of exogenous 15NH4Cl in the gulf toadfish (Opsanus beta). AB - This study was undertaken to determine whether gulf toadfish (Opsanus beta) could metabolize ammonia from their environment into other, less toxic products. To this end, gulf toadfish were exposed to 3.8 mM 15NH(4)Cl in seawater for 24 and 48 h. Liver, kidney, gill, brain and muscle samples were analyzed for distribution of 15N within the tissue and among various nitrogen-containing metabolites (ammonia, amino-N, glutamine-N, urea and protein). The data reported here show that the toadfish can indeed take up and metabolize ammonia. Analysis of individual metabolic products of ammonia indicates that the toadfish can convert this toxic chemical into other less toxic metabolites. Ammonia enrichment is significantly different over controls in the kidney, brain and muscle. Urea enrichment is most significant in the brain, with less significant enrichment occurring in the liver and muscle. While accumulation of ammonia into an amino acid pool was not a significant metabolic fate, protein synthesis was significantly enriched in all tissues (with the highest levels occurring in the gill) indicating that amino acid synthesis may be a pathway of ammonia detoxification en route to protein synthesis, and that environmental ammonia can be 'fixed' into protein. Finally, it was found that glutamine-N synthesis occurs at significant levels in the liver, brain and muscle. PMID- 14559298 TI - The effects of Eucalyptus terpenes on hepatic cytochrome P450 CYP4A, peroxisomal Acyl CoA oxidase (AOX) and peroxisome proliferator activated receptor alpha (PPARalpha) in the common brush tail possum (Trichosurus vulpecula). AB - Eucalyptus leaves contain a high proportion of essential oils comprising of a complex mixture of monoterpenes such as 1,8-cineole, alpha-pinene, d-limonene and p-cymene. In this study, hepatic levels of microsomal lauric acid hydroxylase and peroxisomal cyanide-intensive palmitoyl coenzyme A oxidative activities were examined in livers of possums given an artificial diet consisting of the above monoterpenes for 10 days. These values were compared with those of possums fed a control diet containing only fruits and cereals. Peroxisomal cyanide-intensive palmitoyl coenzyme A oxidative activity was significantly higher in livers of treated possums relative to that of control possums (2.96+/-0.93 vs. 0.98+/-0.88 nmol/mg protein per min, P<0.01) (mean+/-S.D., n=4). A small increase in microsomal lauric acid hydroxylase activity was observed in the treated possum in comparison with the control group (4.40+/-0.85 vs. 3.60+/-0.48 nmol/mg protein per min) (mean+/-S.D., n=4). A higher NAD/ NADP ratio was observed in treated possums as compared with control possums (4.73+/-0.65 vs. 3.51+/-0.64 nmol/mg protein per min, P<0.05) (mean+/-S.D., n=4). No other statistically significant differences in pyridine nucleotide contents were found between control and treated possums. Northern blot analysis of mRNA from rat, human, terpene treated and control possum livers, using the corresponding koala cDNA probes, detected a more intense acyl CoA oxidase (AOX) mRNA band in livers of terpene fed possums. Negligible differences in the intensity of CYP4A and PPARalpha mRNA bands were observed between the two groups. These data suggest that Eucalyptus terpenes elevate hepatic AOX expression in possums. PMID- 14559299 TI - Catechin attenuates 6-hydroxydopamine (6-OHDA)-induced cell death in primary cultures of mesencephalic cells. AB - Past studies have shown the protective effects of tea catechins on oxidative cell damage induced by 6-OHDA in PC12 cells. In this study we verified whether or not catechin prevents 6-OHDA-induced oxidative cell damage in primary cultures of rat mesencephalic cells. On exposure to 6-OHDA (200 microM), the cultures showed a marked decrease in cell viability, disturbances in lipid peroxidation, and an increased generation of NO, as assayed by MTT, TBARS and nitrite assays, respectively. Introduction of catechin significantly attenuated the cell death caused by 6-OHDA at concentrations of 3.4, 34 and 340 microM in a dose-related manner. Catechin produced no marked changes on 6-OHDA-induced increases in NO, but caused a significant inhibition of lipid peroxidation. These results suggest that catechins offer similar cytoprotection against 6-OHDA-induced oxidative cell damage in mesencephalic cell cultures, as previously described in PC12 cells. The cytoprotective function of catechin results from its antioxidant property and is not due to the inhibition of nitric oxide synthase. These findings further support and substantiate traditional consumption of catechin rich green/black tea as protection against neurodegenerative diseases like Parkinsonism. PMID- 14559300 TI - Effect of heavy metals on the uptake of [3H]-L-histidine by the polychaete Nereis succinea. AB - Integumentary uptake of [3H]-L-histidine by Nereis succinea was measured in the presence and absence of selected heavy metals and the amino acid L-leucine in 60% artificial seawater (ASW). The time course of 10 microM [3H]-L-histidine uptake into worms over a 60 min incubation was approximately doubled in the presence of 0.5 microM zinc and when calcium in the incubation medium was reduced from 6 mM to 5 microM the stimulatory effect of zinc on amino acid accumulation was reduced and uptake under the latter conditions was approximately half that of the control. Zinc stimulation of [3H]-L-histidine influx was a hyperbolic function of zinc concentration over the range 0 to 50 microM metal and displayed an apparent activation or affinity constant of 385+/-127 nM Zn(2+). The hyperbolic stimulatory effect of 1 microM Zn(2+) on the time course of 10 microM [3H]-L histidine uptake was abolished in the presence of 25 microM L-leucine, suggesting that this amino acid shared the same transport system as [3H]-L-histidine and acted as a potential competitive inhibitor. Influx of [3H]-L-histidine was a hyperbolic function of external amino acid concentration and displayed an apparent affinity constant (Km) of 23.71+/-5.02 microM and an apparent aximal velocity (J(max)) of 4701+/-449 pmol/g dry wt.x15 min. Addition of 0.5 microM zinc resulted in a four-fold increase in J(max) and a doubling of K(m), suggesting the effect of the metal was mostly on the rate of amino acid transport. [3H]-L-histidine influx was mildly stimulated by Fe(2+) (0.5 microM), but was unaffected by either Ag(+) or Al(3+) (both at 0.5 microM). These results suggest that [3H]-L-histidine uptake into worm integument may take place by the classical Na(+)-independent L-transport system shared by L-leucine and regulated by exogenous calcium and other divalent metal concentrations. PMID- 14559301 TI - [Oesophageal carcinoma: what treatment? Which patients must be operated on?]. PMID- 14559302 TI - [Adenocarcinoma of the gastric stump]. AB - Cancer of the gastric stump is a classical late complication of gastrectomy for benign lesion. This tumor is defined by various criteria, including a minimal delay of 5 years since the initial gastrectomy and the benignity of the initial lesion. Early diagnosis is difficult since suggestive clinical signs are usually associated with advanced tumors. Prognosis is globally bad and theoretically justifies routine endoscopic screening. For tumors which can be radically resected, completion gastrectomy with lymphadenectomy is indicated and allows a 40% 5-year survival. In other cases, palliative treatment remains a major concern. PMID- 14559303 TI - [Polytrauma is more severe after a free fall from a height than after a motor vehicle accident]. AB - AIMS: To compare specific features and prognosis of polytrauma victims requiring urgent laparotomy, after a free fall from a height (FFH) or a motor vehicle accident (MVA). PATIENTS AND METHODS: Urgent laparotomy was performed only in case of massive hemoperitoneum associated with hemodynamic instability. Injury severity score (ISS) and hospital mortality rate were used to determine the severity of trauma. RESULTS: Two hundred and four patients underwent urgent laparotomy for blunt abdominal trauma: 79 after FFH (intentional defenestration: 65, accident: 14), and 125 after a MVA. Mean ISS was higher after FFH (39 +/- 11) than MVA (35 +/- 12) (P< 0.05). Spine injury and pelvic fractures were more frequent and more severe in FFH patients, with a high incidence of retroperitoneal hemorrhage. Juxta-hepatic veins and inferior vena cava tears usually occurred after FFH. Hospital mortality was 48% (38/79) after FFH and 30% (37/125) after MVA (P< 0.01). Predominant cause of death was associated injuries. The height of the fall was not a prognostic factor, unlike the quality of the landing surface: the mortality dropped from 59% (34/58) when patient fell onto a hard ground, to 19% (4/21) in case of soft landing surface (P< 0.05). CONCLUSION: Blunt abdominal trauma requiring urgent laparotomy is more severe after FFH than after MVA. The usual cause of death is associated injuries in both groups. Quality of the landing surface is a discriminating factor for mortality after FFH. PMID- 14559304 TI - [Advantages of using robotic Da Vinci system for unilateral adrenalectomy: early results]. AB - STUDY AIM: The goal of this study was to report the early results of unilateral transperitoneal adrenalectomy using robotic Da Vinci system, and to compare them to the results of the laparoscopic standard adrenalectomy. METHODS: Prospective study included all patients operated on for unilateral laparoscopic or robotic adrenalectomy from November 2000 to November 2002. RESULTS: Twenty-eight patients underwent unilateral adrenalectomy using either standard laparoscopy (14 patients) or robotic Da Vinci system (14 patients). Mean duration of robotic adrenalectomy seemed to be longer than standard laparoscopy (111 vs. 83 min; P = 0.057). This tendency decreased while surgeons' experience was increasing. Mean duration of operating room activity was similar for both types of surgery. Peroperative events without conversion, conversion rate (7%), drainage, morbidity (21%), duration of hospitalisation were similar for both types of surgery. Duration of standard laparoscopic adrenalectomy was positively correlated to patients body mass index. This correlation was absent in patients operated on by robotic Da Vinci system. CONCLUSION: This preliminary study found no objective data demonstrating that robotic Da Vinci system was superior to standard laparoscopic approach for unilateral adrenalectomy. However, we think that it is necessary to continue further evaluation of this system to demonstrate its possible superiority. PMID- 14559305 TI - [Patients surviving 5 years after curative oesophagectomy for oesophageal cancer]. AB - AIM OF THE STUDY: To analyse the clinical and pathological parameters of 5-year survival patients after curative oesophageal resection for cancer and to identify factors predictive of long-term survival. METHODS: The data of 370 patients who underwent oesophagectomy with curative intent from January 1982 for oesophageal squamous cell carcinoma (n = 320) or adenocarcinoma (n = 50) were reviewed. After excluding postoperative deaths (n = 20), these patients were surviving (S group, n = 113) or dead (NS group, n = 237) with a 60-month follow-up. Uni- and multivariate analysis allowed comparison between the two groups. RESULTS: Postoperative mortality and morbidity rates were 4.0% and 37.6%, respectively. Parameters related to 5-year survival were: absence of preoperative malnutrition or dysphagia, transhiatal resection, no reoperation, limited tumour, histological response to neoadjuvant treatment, absence of lymph node capsular invasion, number of invaded lymph nodes < or = 4, invaded lymph node ratio < or = 0.1, absence of tumour recurrence or metachronous primary cancer. On multivariate analysis, factors predictive of 5-year survival were: absence of preoperative dysphagia (P < 0.001), stage 0-I-IIA tumour (P<0.001) and absence of metachronous cancer (P = 0.016). CONCLUSION: Complete surgical resection allows 5-year survival. Factors predictive of long-term survival assessed in preoperative evaluation, dysphagia and tumour stage, should be useful to select patients for neoadjuvant treatment. PMID- 14559307 TI - [When and how to operate a lithiasic acute cholecystitis?]. PMID- 14559306 TI - [Solid-pseudopapillary tumor of the pancreas: a clinical study of five cases, and review of the literature]. AB - INTRODUCTION: Solid pseudopapillary tumors of the pancreas are rare and their origin is unknown. The aim of this work was to report five new cases. MATERIAL AND METHODS: Retrospective study of data from patients operated on from 1983 to 2002 in a university hospital specialized in pancreatic surgery. Patients were identified in a prospectively constituted database of pathologic examinations. RESULTS: Five patients (three men and two women, aged from 15 to 69 years) underwent pancreatectomy for a solid pseudopapillary tumor, which was discovered fortuitously by imaging in three cases. Tumor diameter ranged from 4 to 15 cm. Diagnosis was made preoperatively in only one patient. There were three pancreaticoduodenectomies and two left pancreatectomies, with extension to the transverse colon due to vascular reasons in two cases. Only one significant complication occurred (one colonic fistula). With a follow-up ranging from 6 months to 6 years, all patients are alive without recurrence. CONCLUSIONS: Solid pseudopapillary tumors are not exceptional in men. Complete resection can need extension to neighboring organs but allows good long-term survival. PMID- 14559308 TI - [A case of right liver atrophy]. AB - The authors report a case of right liver atrophy. This rare anomaly was suspected during post-operative period on abnormalities of cholangiogram. This biliary tract anomaly was diagnosed by CT scan. This congenital abnormality may be associated with biliary tract abnormalities, portal hypertension and other congenital abnormalities. PMID- 14559309 TI - [Synovial cyst of the hip: a misleading strangulated crural hernia diagnosis]. AB - We report a case of crural tumefaction becoming rapidly painful revealing a synovial cyst of the hip. Literature data concerning differential diagnosis, pathogenesis, clinical varieties, diagnosis and treatments of this rare pathology are emphasized. PMID- 14559310 TI - [Primary and bilateral non-Hodgkin's lymphoma of the adrenal gland (a case report and literature review)]. AB - Primary non-Hodgkin's lymphoma of the adrenal gland is rare. We report the case of a 31-years-old patient hospitalized with asthenia and adrenal insufficiency. The CT scan showed a bilateral adrenal mass. A scano-guided biopsy suspected an endocrinoid tumor. The surgical exploration demonstrated a huge mass invading the retroperitoneal space, and the biopsy concluded to a central follicular phenotype B rmalignant lymphoma with a high rank of malignity. The thoracic CT scan did not show any lymph node. The medullar biopsy eliminated a secondary lymphoma. The patient was treated by chemotherapy and radiotherapy with a good result during 16 months. PMID- 14559311 TI - [Spontaneous cervical haematoma due to extracapsular haemorrhage of a parathyroid adenoma: a report of 2 cases]. AB - The usual clinical manifestations of a parathyroid adenoma are due, in most of the cases, to hypercalcemia. The development of a spontaneous cervical or cervicomediastinal haematoma is a rare form of presentation. In case of a spontaneous cervical haematoma associated with dysphagia: measurement of serum calcium, phosphate and parathyroid hormone allows the diagnosis of haematoma due to extracapsular haemorrhage from a parathyroid adenoma. We report herein 2 cases. PMID- 14559312 TI - [Umbilical hernias treatment by the "stamp mesh technique"]. AB - We describe an original technique for umbilical hernia repair ("stamps mesh technique") during which no umbilical ring widening is carried out. A non absorbable mesh is positioned in the preperitoneal space. PMID- 14559313 TI - [Surgical management of mediastinitis and mediastinal abscess: a series of 56 cases]. PMID- 14559314 TI - Prevalence of hospital malnutrition in Latin America: the multicenter ELAN study. AB - OBJECTIVE: We determined the nutrition status and prevalence of malnutrition as determined by the Subjective Global Assessment in Latin America, investigated the awareness of the health team with regard to nutrition status, evaluated the use of nutritional therapy, and assessed the governmental policies regulating the practice of nutritional therapy in each country. METHODS: This cross-sectional, multicenter epidemiologic study enrolled 9348 hospitalized patients older than 18 y in Latin America. Student's t test and chi-square tests were used to analyze univariate analysis and multiple logistic regression analysis, respectively. RESULTS: Malnutrition was present in 50.2% of the patients studied. Severe malnutrition was present in 11.2% of the entire group. Malnutrition correlated with age (>60 y), presence of cancer and infection, and longer length of hospital stay (P < 0.05). Fewer than 23% of the patients' records contained information on nutrition-related issues. Nutritional therapy was used in 8.8% of patients (6.3% enteral nutrition and 2.5% parenteral nutrition). Governmental policies ruling the practice of nutritional therapy exist only in Brazil and Costa Rica. CONCLUSIONS: Hospital malnutrition in Latin America is highly prevalent. Despite this prevalence, physicians' awareness of malnutrition is weak, nutritional therapy is not used routinely, and governmental policies for nutritional therapy are scarce. PMID- 14559315 TI - Plasma total homocysteine in Mexican rural and urban women fed typical model diets. AB - OBJECTIVE: The purpose of the present work was to determine the fasting plasma total homocysteine (tHcy) levels and the time-course response of tHcy concentrations after the consumption of urban and rural Mexican model diets in two groups of Mexican women from urban and rural areas. METHODS: Thirty-three adult women (age range = 18-49 y) were studied. Fifteen women were from a rural community in the state of Mexico. The other 18 were from cities and consumed diets that regularly included an important amount of animal foods. The study was designed as a two-period crossover study in which subjects consumed the model urban or rural diet in a 2-wk interval. Seven milliliters of venous blood was drawn before ingestion of experimental diets (basal) to measure total cholesterol, high-density lipoprotein cholesterol, triacylglycerol, tHcy, folate, vitamin B12, and methionine. Blood samples were then obtained 30, 60, 90, 180, and 240 min after the beginning of meal consumption. RESULTS: The rural and urban groups showed similar concentrations of tHcy 4 h after meal consumption and after fasting. However, the urban and rural groups had higher methionine plasma concentrations after the urban diet than after the rural diet. In contrast, there was no significant difference in methionine plasma levels between the rural and urban groups with each diet. Those women with low tHcy concentrations maintained those values over the study period, and those with high tHcy concentrations maintained those values. There was no significant difference in tHcy concentrations due to consumption of the two diets (P = 0.31) or the interaction between population and diet (P = 0.84). However, there was a significant difference in the concentration of tHcy between the rural (8.73 +/- 0.17 microM/L) and the urban (9.27 +/- 0.13 microM/L) populations (P = 0.01). In both groups, average tHcy concentration was in the normal range. In both populations, the nutrition status for folate and vitamin B12 was adequate, although plasma folate concentration was significantly lower in the rural population than in the urban population (P < 0.01). Plasma vitamin B12 concentrations were similar in both groups. No subject had low plasma vitamin B12. CONCLUSIONS: Plasma tHcy concentrations in rural and urban Mexican women were within the range considered adequate; however, urban women showed significant higher concentrations than did rural women independently of the consumed diet and the plasma methionine concentration. These results indicated that there is no short-term variation in plasma tHcy due to the consumption of rural or urban diets. PMID- 14559316 TI - Carbohydrate malabsorption may increase daily energy requirements in infants. AB - OBJECTIVE: Carbohydrate malabsorption in infants has been found to increase nutrient losses. However, the effect of this alteration on daily metabolic rate is unknown. We assessed daily metabolic rates in infants with asymptomatic carbohydrate malabsorption (ACM) after a single fruit juice load. METHODS: Sixteen healthy infants with ACM (63.3 +/- 5.6 cm, 7.5 +/- 1.0 kg, 5.6 +/- 0.8 mo, peak breath hydrogen [BH2] = 39.1 +/- 22.4 ppm) and 16 without ACM (64.3 +/- 3.9 cm, 7.8 +/- 1.0 kg, 5.0 +/- 0.8 mo, BH2 = 9.4 +/- 4.7 ppm), after a single fruit juice load, had 24-h energy expenditure (24-h EE; kcal x kg(-1) x d(-1)), resting (RMR; kcal x kg(-1) x d(-1)) and sleeping (SMR; kcal x kg(-1) x d(-1)) metabolic rates extrapolated from 3.5-h assessments in the Enhanced Metabolic Testing Activity Chamber. Furthermore, RMR was calculated with the World Health Organization (WHO), Schofield weight-based and weight- and height-based equations. Carbohydrate absorption was determined by BH2. Differences (P < 0.05) were determined by t test. RESULTS: All infants with ACM had greater (P < 0.05) extrapolated 24-h EE (91.2 +/- 24.8 versus 78.0 +/- 6.8) and RMR (71.8 +/- 15.2 versus 59.5 +/- 5.9). This represented an increase of 15-18.5%, respectively, in energy expenditures. Carbohydrate malabsorption was a significant determinant of EE, RMR, and SMR. However, the WHO (53.8 +/- 1.0 versus 54.1 +/- 0.9) and both Schofield equations (54.7 +/- 0.9 versus 54.9 +/- 1.0 and 50.6 +/- 7.5 versus 47.3 +/- 6.7) failed to detect any differences in RMR. There was a 20 percentile reduction in growth performance in infants with carbohydrate malabsorption. CONCLUSIONS: Infants with ACM following fruit juice ingestion may have increased daily energy expenditure leading to increased metabolic requirements. PMID- 14559317 TI - Decreased oxidative stress in patients with ulcerative colitis supplemented with fish oil omega-3 fatty acids. AB - OBJECTIVE: The potential pathogenicity of free radicals may have a pivotal role in ulcerative colitis. Fish oil omega-3 fatty acids exert anti-inflammatory effects on patients with ulcerative colitis (UC), but the precise mechanism of the action of fish oil on oxidative stress is still controversial. The aim of the present work was to verify the blood oxidative stress in patients with UC and determine whether the association of sulfasalazine to fish oil omega-3 fatty acids is more effective than isolated use of sulfasalazine to reduce the oxidative stress. METHODS: Nine patients (seven female and two male; mean age = 40 +/- 11 y) with mild or moderate active UC were studied in a randomized crossover design. In addition to their usual medication (2 g/d of sulfasalazine), they received fish oil omega-3 fatty acids (4.5 g/d) or placebo for 2-mo treatment periods that were separated by 2 mo, when they only received sulfasalazine. Nine healthy individuals served as control subjects to study the oxidative stress status. Disease activity was assessed by laboratory indicators (C-reactive protein, alpha1-acid glycoprotein, alpha1-antitrypsin, erythrocyte sedimentation rate, albumin, hemoglobin, and platelet count), sigmoidoscopy, and histology scores. Analysis of oxidative stress was assessed by plasma chemiluminescence and erythrocyte lipid peroxidation, both induced by tert butyl hydroperoxide (t-BuOOH) and by plasma malondialdehyde. Antioxidant status was assayed by total plasma antioxidant capacity (TRAP) and microsomal lipid peroxidation inhibition (LPI). Superoxide dismutase (SOD) and catalase erythrocyte enzymatic activities were also determined. RESULTS: No significant changes were observed in any laboratory indicator or in the sigmoidoscopy or histology scores, with the exception of erythrocyte sedimentation rate, which decreased with both treatments. Oxidative stress was demonstrated by significant decreases in TRAP and LPI levels, increased chemiluminescence induced by t-BuOOH, and higher SOD activity in patients with UC. Treatment with fish oil omega-3 fatty acids reverted the chemiluminescence induced by t-BuOOH and LPI to baseline levels but that did not occur when patients received only sulfasalazine. Levels of plasma malondialdehyde, erythrocyte lipid peroxidation, and catalase were not different from those in the control group. CONCLUSIONS: The results indicated that plasma oxidative stress occurs in patients with UC, and there was a significant decrease when the patients used sulfasalazine plus fish oil omega-3 fatty acids. However, there was no improvement in most laboratory indicators, sigmoidoscopy, and histology scores. The results suggested that omega-3 fatty acids may act as free radical scavengers protecting the patients against the overall effect of oxidative stress. PMID- 14559318 TI - Bedside placement of small bowel feeding tubes in hospitalized patients: a new role for the dietitian. AB - OBJECTIVE: The benefits of enteral nutrition when compared with parenteral nutrition are well established. However, provision of enteral nutrition may not occur for several reasons, including lack of optimal feeding access. Gastric feeding is easier to initiate, but many hospitalized patients are intolerant to gastric feeding, although they can tolerate small bowel feeding. Many institutions rely on costly methods for placing small bowel feeding tubes. Our goal was to evaluate the effectiveness of a hospital-developed protocol for bedside-blind placement of postpyloric feeding tubes. METHODS: The Surgical Nutrition Service established a protocol for bedside placement of small bowel feeding tubes. The protocol uses a 10- or 12-French, 110-cm stylet containing the feeding tube; 10 mg of intravenous metoclopramide; gradual tube advancement followed by air injection and auscultation; and an abdominal radiograph for tube position confirmation. In a prospective manner, consults received by the surgical nutrition dietitian for feeding tube placements were followed consecutively for a 10-mo period. The registered dietitian recorded the number of radiograph examinations, the final tube position, and the time it took to achieve tube placement. RESULTS: Because all consults were included, feeding tube placements occurred in surgical and medical patients in the intensive care unit and on the ward. Of the 135 tube placements performed, 129 (95%) were successfully placed postpylorically, with 84% (114 of 135) placed at or beyond D3. Average time for tube placement was 28 min (10 to 90 min). One radiograph was required for 92% of the placements; eight of 135 (6%) required two radiographs. No acute complications were associated with the tube placements. CONCLUSIONS: Hospitalized patients can receive timely enteral feeding with a cost-effective feeding tube placement protocol. The protocol is easy to implement and can be taught to appropriate medical team members through proper training and certification. PMID- 14559319 TI - Soluble tumor necrosis factor-alpha receptor type 1 during selenium supplementation in psoriasis patients. AB - OBJECTIVE: Tumor necrosis factor-alpha (TNF-alpha) and its receptors play important roles in the induction and maintenance of psoriatic lesions. Selenium (Se), a trace element with immunomodulatory properties, is usually decreased in psoriasis patients. We examined the influence of Se supplementation on soluble TNF-alpha receptor type 1 (sTNF-R1) and topical treatment in psoriasis patients. METHODS: The study was conducted in between January and June 2002. Twenty-two inpatients with active plaque psoriasis received topical treatment with 5% salicylic acid ointment, 0.1% to 0.3% dithranol ointment, and 200 microg daily of Se as selenomethionine (SeMet; n = 11, group 1) or placebo (n = 11, group 2) for 4 wk. Psoriasis Area and Severity Index (PASI) score and Se and sTNF-R1 concentrations were assessed at baseline and every 2 wk. Control sera were obtained from 10 healthy subjects. For statistical analysis, parametric tests were used, and the level of significance was set at P = 0.05. RESULTS: The baseline sTNF-R1 levels were 1.87 +/- 0.58 ng/mL (1.98 +/- 0.44 ng/mL in group 1 and 1.75 +/- 0.69 ng/mL in group 2, P = 0.34) in psoriasis patients and 1.65 +/- 0.25 ng/mL in control subjects (P = 0.17); baseline Se concentrations were 48.31 +/- 13.20 microg/L (48.31 +/- 13.20 microg/L in group 1 and 50.35 +/- 13.49 microg/L in group 2, P = 0.41) in psoriasis patients and 58.30 +/- 17.21 microg/L in control subjects (P = 0.05). A positive correlation between PASI and sTNF-R1 was noticed (r = 0.36, P = 0.04; r = 0.51 in group 1 and r = 0.18 in group 2). After 4 wk, almost complete remission of skin lesions was achieved in both groups, but the PASI score was higher in group 1 than in group 2 (4.30 +/- 3.92 and 1.67 +/- 1.17, respectively; P < 0.05). TNF-R1 levels were 1.81 +/- 0.42 ng/mL in group 1 and 1.33 +/- 0.40 ng/mL in group 2 (P = 0.01), and the correlation between PASI score and TNF-R1 level became inverse (r = -0.24 in group 1 and r = -0.59 in group 2). Se concentrations were 107.51 +/- 18.08 microg/L in group 1 and 56.83 +/- 15.32 microg/L in group 2 (P < 0.01). CONCLUSIONS: Increased level of sTNF-R1 may be an indicator of active psoriasis. Supplementation with selenomethionine was ineffective as adjuvant treatment in plaque psoriasis and may contribute to the maintenance of elevated TNF-R1 concentration in psoriasis patients despite the remission of skin lesions. PMID- 14559320 TI - Validity and accuracy of regional bioelectrical impedance devices to determine whole-body fatness. AB - OBJECTIVE: Growing emphasis on obesity as a risk factor for chronic diseases and commercial availability of impedance devices for the at-home assessment of body fatness have stimulated the need for a critical evaluation of the validity of these instruments. This study determined the reproducibility and accuracy of two commercial impedance devices that use upper (hand-to-hand) or lower (foot-to foot) body contact electrode placements in adults with a wide range of body fatness. METHODS: Body composition was assessed with dual x-ray absorptiometry in apparently healthy adults (62 women and 48 men) ages 21 to 60 y, with a range in body mass index of 18.6 to 40.5 kg/m2. Variability in body fatness predicted with the regional body impedance devices was determined in 10 adults on 5 consecutive d. A 50-kHz, tetrapolar bioelectrical impedance plethysmograph with surface electrode placements on the upper and lower limbs was used to determine reference regional and whole-body impedance values. RESULTS: Variability in body mass (1%) over 5 d was less than body fatness predicted with the upper (2-10%) and lower (3 5%) body devices. Regional and whole-body impedance values were different (P < 0.05) in the women, whereas upper and lower body values were lower (P < 0.05) than whole-body impedance in the men. Dual x-ray absorptiometric determinations of body fatness were similar to predictions based on models derived from physical characteristics (age, stature, body mass, and sex) but significantly different (P < 0.05) from estimates from the impedance devices, which underestimated body fatness. Bias in predictions of body fatness with the regional devices was systematically (P < 0.0001) related to body fatness. CONCLUSION: Use of regional impedance devices to assess body fatness is limited by a lack of precision and accuracy. PMID- 14559321 TI - Comparison of bioelectrical impedance prediction equations for fat-free mass in a population-based sample of 75 y olds: the NORA study. AB - OBJECTIVE: We evaluated the performance of different prediction equations to estimate fat-free mass (FFM) from bioelectrical impedance analysis (BIA) in the elderly. METHODS: This study was based on 106 (51 male and 55 female) free-living 75-y-old subjects who participated in the Goteborg part of the Nordic Research on Ageing (NORA) study during 1991 and 1992. FFM predicted from BIA (FFM(GOT)) was validated against FFM estimated from measurements of total body water and total body potassium (FFM(REF)). FFM was calculated from BIA prediction equations for the elderly developed by Deurenberg et al. (FFM(WAG)) and Roubenoff et al. (FFM(FHS)). FFM also was calculated from an equation developed in subjects with a wide age range by Kyle et al. (FFM(GEN)). Bland-Altman analysis was performed to compare FFM(REF) with FFM(GOT), FFM(WAG), FFM(FHS), and FFM(GEN), respectively. FFM(GOT) also was compared with FFM derived from these published equations. RESULTS: Compared with FFM(REF), the FFM(FHS) and FFM(WAG) underestimated FFM by 2.6 and 7.9 kg in males and 4.2 and 9 kg in females, respectively. The FFM(GEN) underestimated FFM in females by 1.3 kg but not in males (mean difference, -0.04 kg). FFM calculated from the BIA equation developed in this population (FFM(GOT)) neither underestimated nor overestimated FFM as compared with FFM(REF), as expected. The differences between FFM(GOT) and FFMs predicted from these equations were of the same magnitude as that observed with FFM(REF). CONCLUSION: Different prediction equations produced different values for FFM. The age specific equations developed in other populations underestimated FFM, whereas FFM(GEN) produced an unbiased estimate of FFM in males but not in females. Thus, the BIA prediction equation needs to be developed and validated in the population under study. PMID- 14559322 TI - Barriers to adequate nutrition in critically ill children. AB - OBJECTIVE: The study assessed the adequacy of nutrition support in critically ill infants and children and identifies barriers impeding the delivery of estimated energy requirement (EER). METHODS: Forty-two children (median age, 6.6 mo; range, 0-198) who were admitted to a tertiary-level pediatric intensive care unit (PICU) were studied prospectively over a 6-mo period. Patients staying in the PICU longer than a full 3 d and who received enteral or a combination of enteral and parenteral nutrition were eligible for inclusion. Patients were assigned to one of two groups: patients after cardiac surgery (n = 18) and all other diagnoses (n = 24). EERs were compared with actual energy intake, and clinical information was collected throughout the PICU admission. RESULTS: Patients in the PICU received a median of 37.7% (range, 0.2-130.2%) of their EERs. The cardiac group achieved significantly lower energy intakes than did the non-cardiac group (P = 0.02). Only 22 of 42 patients (52%) achieved full EERs at any time during their admission, and this was more likely in non-cardiac patients (67% versus 33%, P = 0.03) Children undergoing cardiac surgery had a significant fall in weight-for age Z scores (WAZ) from PICU admission to discharge (median WAZ, -1.44 versus 2.14; P < 0.001). In both groups, the major barrier to achieving EER was fluid volume restriction. Interruption of feeding for procedures and feeding intolerance reduced energy intake to a lesser degree. CONCLUSIONS: This study highlights the inadequacy of nutrition support in critically ill children in the PICU. Restriction of fluid intake was the main barrier to the delivery of adequate nutrition, particularly in infants undergoing cardiac surgery. PMID- 14559323 TI - In vitro analysis of the properties of Beiqishen tea. AB - OBJECTIVE: Chinese Beiqishen tea was studied in an in vitro test system. METHODS: Phytochemical screening, trace element analysis, and the analysis of antioxidant properties were carried out. Characteristic constituents were determined by chromatographic (capillary gas chromatography and GCQ Ion Trap mass spectrometry) and spectrometric (ultraviolet and UV-VIS) methods. Element concentrations were determined by inductively coupled plasma optical emission spectrometry. Antioxidant capacity was studied by spectrophotometric and luminometric techniques using a Berthold Lumat 9501 luminometer. Hydrogen-donating activity, reducing power, and total scavenger capacity were measured. RESULTS: Total polyphenol content was 20.77 +/- 0.52 g/100 g of drug; total flavonoid content was 0.485 +/- 0.036 g/100 g of drug; and tannin content was 9.063 +/- 0.782 g/100 g of drug. Caffeine content was 1.08 mg/100 g of drug. Essential oils were identified by gas chromatography: (+)-limonene (21%), p-cymene (1.7%), estragol (3.2%), beta-ocimene (1.4%), and thymol (2.6%). Metallic ion analysis showed significantly high concentrations of Al, As, Ba, Cr, Cu, Fe, Mn, Ni, and Ti in the drug. Antioxidant and scavenger properties were identified as a function of concentration. CONCLUSIONS: The tea infusion contained some non-desirable trace elements and caffeine in addition to polyphenols and tannins in high concentrations. Therefore, the consumption of this tea may involve risks. PMID- 14559324 TI - Inhibitory effects of grape seed extract on lipases. AB - OBJECTIVE: The aim of the present study was to assess the effects of grape seed extract (GSE) on the fat-metabolizing enzymes pancreatic lipase, lipoprotein lipase, and hormone-sensitive lipase in vitro and evaluate its potential application as a treatment for obesity. METHODS: Crushed grape seeds were extracted in ethanol, and the extract was assayed for the measurement of inhibitory effects on pancreatic lipase and lipoprotein lipase activities and on lipolysis of 3T3-L1 adipocytes. RESULTS: The GSE rich in bioactive phytochemicals showed inhibitory activity on the fat-metabolizing enzymes pancreatic lipase and lipoprotein lipase, thus suggesting that GSE might be useful as a treatment to limit dietary fat absorption and the accumulation of fat in adipose tissue. The observed reduction in intracellular lipolytic activity of cultured 3T3-L1 adipocytes may reduce the levels of circulating free fatty acids that have been linked to insulin resistance in obese patients. CONCLUSION: The GSE rich in compounds that inhibit lipases may provide a safe, natural, and cost-effective weight control treatment. PMID- 14559325 TI - Effects of dietary glutamine on antioxidant enzyme activity and immune response in burned mice. AB - OBJECTIVES: We investigated the effect of dietary glutamine (Gln) on specific antibody production and antioxidant enzyme activities in burned mice vaccinated with detoxified Pseudomonas exotoxin A linked with the outer membrane proteins I and F (PEIF). We also evaluated the survival rate of vaccinated and non vaccinated burned mice infected with Pseudomonas aeruginosa. METHODS: There were three consecutive experiments. In experiment 1, 30 BALB/c mice were assigned to one of two groups. The control group was fed casein as the protein source; the Gln group received 4% Gln (w/w) to replace part of the casein. Mice were immunized twice with PEIF, and the production of specific antibodies against PEIF was measured every week. Eight weeks after immunization, all mice received a 30% body surface area burn injury. Mice were killed 24 h after the burn. The antioxidant enzyme activities and lipid peroxides in the tissues and specific antibody production were analyzed. In experiment 2, 12 mice were assigned to a control or a Gln group and fed with one the experimental diets for 4 wk. Then burn injury was induced, and mice were killed 24 h later. In vitro, splenocytes were cultured, and interleukin (IL)-4 and IL-10 were measured after mitogen stimulation. In experiment 3, survival rates of vaccinated and non-vaccinated burned mice complicated with P. aeruginosa infection were evaluated. The survival rate was observed for 8 d after the burn. RESULTS: Antioxidant enzyme activities and lipid peroxides in tissues tended to be lower in the Gln group than in the control group after the burn. Specific antibody production against P. aeruginosa increased significantly in the Gln group at 4 and 7 wk after immunization and at 24 h after the burn. IL-4 concentrations in mitogen-stimulated splenocytes were significantly higher in the Gln group than in the control group. Survival rates of non-vaccinated burned mice in the Gln group were significantly higher than those in the control group, whereas there was no difference in the survival of vaccinated burned mice after bacterial infection. CONCLUSIONS: These results suggested that vaccinated mice receiving a Gln-enriched diet may have enhanced humoral immunity and attenuated oxidative stress induced by burn injury. Also, Gln supplementation improved the survival of burned mice complicated with P. aeruginosa infection. PMID- 14559326 TI - Stability of total nutrient admixtures in reference to ambient temperatures. AB - OBJECTIVE: To evaluate the stability of emulsions under different temperatures simulating clinical conditions of storage and exposure during infusion, five total nutrient admixture formulas in this institution were analyzed: adult, patients with hepatic failure, infants, stressed patients, and patients with renal failure. METHODS: Each mixture was allocated in a sterile 100-mL glass bottle, which was prefilled and refilled with nitrogen gas. Bottles were stored at 4 degrees C for 0 d, 3 d, and 7 d and then exposed to three different temperatures: usual room temperature (18 degrees C to 25 degrees C), high (>28 degrees C) in a water bath, or storage (4 degrees C) for 24 and 48 h. The gross inspection of the emulsions and parallel measurements of pH, particle sizes, divalent ions, peroxide levels, and microbial cultures were performed. RESULTS: Every lot was stable near 18 degrees C, but 8 of 10 lots stored for 7 d (25 degrees C and >28 degrees C) and 15 of 20 lots stored for 3 and 7 d (25 degrees C and >28 degrees C) showed coalescence. The overall coalescence incidences by storage, exposure, and heat were statistically significant (P < 0.005). CONCLUSION: For the safety of total nutrient admixtures, special attention is required to keep the ambient temperature below 28 degrees C and completely exclude air from the container. PMID- 14559327 TI - Effect of endurance training on muscle fat metabolism during prolonged exercise: agreements and disagreements. PMID- 14559328 TI - Ketogenic diet modifies the risk factors of heart disease in obese patients. PMID- 14559330 TI - Delayed presentation of a gastric colocutaneous fistula after percutaneous endoscopic gastrostomy. PMID- 14559329 TI - Pravastatin in hyperlipidemia secondary to HIV protease inhibitors without response to fenofibrate: a brief preliminary report. PMID- 14559331 TI - Starving in the hospital. PMID- 14559333 TI - Comparison of FAIR technique with different inversion times and post contrast dynamic perfusion MRI in chronic occlusive cerebrovascular disease. AB - The purpose of this study was to examine the signal change occurring with different inversion times (TIs) of the flow-sensitive alternating inversion recovery (FAIR) technique and to compare with the perfusion image obtained with Gd-DTPA injection. The subjects were 11 patients with unilateral occlusive cerebrovascular disease. Two FAIR images with different TIs (800 ms and 1600 ms) were measured for each patient and dynamic perfusion MRI was performed to produce four kinds of parameter maps: mean transit time (MTT), time to peak (TTP), relative cerebral blood flow (rCBF) and relative cerebral blood volume (rCBV) maps. Asymmetry ratios (ARs) between the affected and contra-lateral vascular sides were measured in both FAIR images and the four dynamic parameter maps. The AR of the MTT map of the four parameters showed the highest correlation with that of the FAIR images, especially with that of TI = 1600 ms (r = 0.829), and the AR of the rCBV map revealed the worst correlation with the FAIR images. The AR of the FAIR image with TI = 800 ms was less correlated with that of MTT than that with TI = 1600 ms. These results suggested that the signal intensity of the FAIR image was influenced by flow transition time and the change in TI could be used to select the flow with a different transition time. Our study suggested that a longer TI in the FAIR technique might be more useful than a shorter TI for evaluating chronic occlusive cerebrovascular disease in the clinical setting. PMID- 14559332 TI - Superparamagnetic iron oxide (SPIO) enhancement in the cirrhotic liver: a comparison of two doses of ferumoxides in patients with advanced disease. AB - The aim of this study was to establish whether enhancement of the liver by the MRI contrast agent ferumoxides could be effectively achieved at a reduced dose of 7.5 micromol/kg in patients with advanced liver cirrhosis. Forty-two liver transplant candidates with end-stage cirrhosis underwent SPIO-enhanced MRI at 1.5T, using either 15 micromol/kg or 7.5 micromol/kg ferumoxides. The lower dose of ferumoxides was also used in 21 non-cirrhotic patients with colorectal liver metastases who acted as a control group. The percentage signal intensity loss (PSIL) after SPIO was measured in all patients, and in those patients with tumors the post-SPIO contrast-to-noise ratio (CNR) was measured. The median PSIL after SPIO in the high dose cirrhotic (HDLC), low dose non-cirrhotic (LDNC) and low dose cirrhotic (LDLC) patients was 86.3%, 74.6%, and 64.2% respectively. These differences were significant using the Mann-Whitney U test. Tumors were found in 8 patients in the high dose cirrhotic group, 9 in the low dose cirrhotic group, and all 21 of the control group. No significant differences were found between the CNR values after SPIO in the 3 groups (median values HDLC 15.1, LDNC 23.7, LDLC 19.5). In patients with late-stage cirrhosis the PSIL after SPIO was significantly less at 7.5 micromol/kg than at 15 micromol/kg, but both doses produced a substantial loss of signal. Lesion to liver CNR was not adversely affected by using the lower dose, so when imaging at 1.5T the authors would recommend using 7.5 micromol/kg in patients with liver cirrhosis. PMID- 14559334 TI - Inter-rater and intra-rater reliability of subchondral bone and cartilage thickness measurement from MRI. AB - MRI is often used to visualize and quantify the articular cartilage layer of load bearing joints affected by degenerative diseases, such as osteoarthritis (OA). Although the role played by the subchondral bone in the etiology and/or progression of OA may be important, the ability to visualize and quantify subchondral bone with MRI has received little attention. In this report we examined the inter-rater and intra-rater reliability of subchondral bone and cartilage thickness measurements from MR images of cadaver femoral head specimens. A 3D-SPGR pulse sequence tuned to eliminate chemical shift artifact through phase cancellation was used to image the specimens. Three raters manually segmented four specimens on two different occasions. Subchondral bone and cartilage thickness measurements were calculated from the segmented images. Inter rater and intra-rater reliabilities were very high (>.98) for both cartilage and subchondral bone thickness measurements. We conclude that subchondral bone thickness can be measured as reliably as cartilage thickness from MR images. PMID- 14559335 TI - MRI-induced stimulation of peripheral nerves: dependency of stimulation threshold on patient positioning. AB - Fast gradient systems enable the imaging of moving organs but also restrict imaging possibilities by stimulating the peripheral nerves. Several studies have determined the stimulation thresholds of the peripheral nerves. However, no study has explicitly examined whether stimulation thresholds are dependent on the position of the patient in the MR imager. In this study, combinations of one, two and three gradient axes were switched with sinusoidal wave forms (rise time 300 micros). Stimulation thresholds were determined for five patient positionings: gradient isocenter at the bridge of the nose, jugular fossa, 10 cm cranial of the umbilicus, umbilicus, and 10 cm caudal of the umbilicus. Experiments were performed in two volunteer groups, one for triple gradient switching (17 male, 14 female) and the other for single and double gradient switching (14 male, 11 female). The dependence of the stimulation threshold on the position of the volunteers in the MR imager was statistically significant for most of the gradient combinations, with the highest stimulation thresholds for positions umbilicus+10 and umbilicus at isocenter. Similar patterns of position dependence were obtained for gradient pairs whose axes were axially symmetric to the human body. Mean stimulation thresholds of male volunteers were lower than those of female volunteers. These findings indicate that stimulation thresholds should be defined according to the position of the patient in the MR scanner. PMID- 14559336 TI - MRI of blood volume with MS 325 in experimental choroidal melanoma. AB - Functional magnetic resonance imaging (MRI) allows quantitative blood volume imaging in vivo at high tissue resolution. The purpose is to apply this technique for untreated and hyperthermia-treated experimental choroidal melanoma. MS 325 was used as new intravascular albumin-bound gadolinium-based contrast agent. Pigmented choroidal melanomas were established in albino rabbits. MRI was performed in 7 untreated eyes and 7 eyes treated with a Neodymium:Yttrium Lanthanum-Fluoride-laser at 1047 nm. 3D-spoiled gradient echo pulse sequences were used to acquire T' weighted axial images. First, a set of images was collected without contrast agent. MS 325 was then injected i.v. and images were obtained within 12 min after injection. Signal intensities were measured within tumor, ciliary body, choroid, and iris and relative signal intensities were determined for these tissues in relation to vitreous. In untreated tumors, the relative signal intensity was higher after injection of MS 325 (5.61+0.70) than without MS 325 (2.90+0.33; p = 0.0002). In contrast, the relative signal intensity of treated tumors did not differ significantly before and after MS 325 (6.19+1.59 and 6.13+1.64). Histopathological sections indicated vascular occlusion in treated tumors. All other studied tissues of untreated and treated eyes showed a significant increase of relative signal intensities in the presence of MS 325. An animal model for the research on contrast agents in MRI is presented. Blood volume measurement with MS 325 was adapted for experimental choroidal melanomas. Reduced change of relative signal intensity indicates compromised blood volume after vascular occlusion in hyperthermia-treated melanoma. Further studies are needed to investigate whether this technique allows the evaluation of tumor viability following treatments. PMID- 14559337 TI - Intraoperative contrast-enhanced MR-imaging as predictor of tissue damage during cryoablation of porcine liver. AB - This study evaluate intraoperative Magnetic Resonance Imaging (MRI) as predictor of tissue damage following cryoablation of porcine liver with and without concomitant hepatic vascular inflow occlusion. Inflow occlusion was used during freezing in 6 of 12 pigs included. The volumes of the procedural ice-balls were estimated from MR images. Immediately after thawing contrast (MnDPDP) enhanced MRI was performed to estimate the volume of the cryolesion. Four days after ablation MRI was repeated of the in-vivo and the ex-vivo liver. Photography was performed of the sliced liver specimens to estimate the volumes of the lesions. The intraoperative volume of the cryolesion as shown by contrast enhanced MRI corresponded well to the ice-ball volume for lesions made without vascular occlusion (difference 0.3 +/- 0.9 cm(3), p = 0.239). For lesions made during occlusion the volume of the intraoperative cryolesion was larger than the corresponding ice-ball (difference 7.5 +/- 3.3 cm(3), p = 0.003). The volume of the cryolesions as estimated from histopathology four days after freezing and contrast enhanced MRI immediately after freezing corresponded well for lesions made with (difference -2.6 +/- 4.5 cm(3), p = 0.110) and without vascular occlusion (difference -0.5 +/- 2.3 cm(3), p = 0.695). Intraoperative MnDPDP enhanced MRI of the cryolesion is predictive of the tissue damage induced during cryoablation of porcine liver. The procedural ice-ball is not, if induced during inflow occlusion. PMID- 14559338 TI - Manifestation of magic angle phenomenon: comparative study on effects of varying echo time and tendon orientation among various MR sequences. AB - The purpose of this study was to systematically investigate the effect of varying the echo time (TE) values and angle of the tendon to the main magnetic field (B(o)) upon the signal intensity observed with the magic angle phenomenon in tendons among most commonly used MR pulse sequences, including conventional spin echo (CSE), fast spin echo (FSE) and gradient echo (GRE) sequences. The intact bovine Achilles tendon was imaged using a clinical 1.5-T MR scanner. Magic angle phenomenon occurs in CSE, FSE and GRE sequences with different grade, appearing most severe in CSE, middle in FSE, and weakest in GRE sequence. In addition, the tendon signal changes produced by the magic angle phenomenon could be greatly reduced by increasing the TE to above a certain critical value in all three sequences. These critical TE values were different among CSE (40 msec), FSE (70 msec), and GRE (30 msec) sequences. PMID- 14559339 TI - Fast T2-weighted MR imaging: impact of variation in pulse sequence parameters on image quality and artifacts. AB - The purpose of this study was to quantitatively evaluate in a phantom model the practical impact of alteration of key imaging parameters on image quality and artifacts for the most commonly used fast T(2)-weighted MR sequences. These include fast spin-echo (FSE), single shot fast spin-echo (SSFSE), and spin-echo echo-planar imaging (EPI) pulse sequences. We developed a composite phantom with different T1 and T2 values, which was evaluated while stationary as well as during periodic motion. Experiments involved controlled variations in key parameters including effective TE, TR, echo spacing (ESP), receive bandwidth (BW), echo train length (ETL), and shot number (SN). Quantitative analysis consisted of signal-to-noise ratio (SNR), image nonuniformity, full-width-at-half maximum (i.e., blurring or geometric distortion) and ghosting ratio. Among the fast T(2)-weighted sequences, EPI was most sensitive to alterations in imaging parameters. Among imaging parameters that we tested, effective TE, ETL, and shot number most prominently affected image quality and artifacts. Short T(2) objects were more sensitive to alterations in imaging parameters in terms of image quality and artifacts. Optimal clinical application of these fast T(2)-weighted imaging pulse sequences requires careful attention to selection of imaging parameters. PMID- 14559340 TI - Three dimensional k-space trajectory design using genetic algorithms. AB - Image quality and total scan time in MRI are determined in large part by the trajectory employed to sample the Fourier space. Each trajectory has different properties like coverage of k-space, scan time, sensitivity to off-resonance conditions, etc. These properties are often contradictory, therefore a universal optimal trajectory does not exist and ultimately, it will depend on the image characteristics sought. Most trajectories used today are designed based on intuition and k-space analysis more than with optimization methods. This work presents a 3D k-space trajectory design method based on Genetic Algorithm optimization. Genetic Algorithms have been chosen because they are particularly good for searching large solution spaces. They emulate the natural evolutionary process allowing better offsprings to survive. The objective function searches the maximum of the trajectory's k-space coverage subject to hardware constraints for a fixed scanning time using the trajectory's torsion as its optimization variable. The method proved to be effective for generating k-space trajectories. They are compared with well-established trajectories. The results of simulated experiments show that they can be appropriate for image acquisition under certain special conditions, like off-resonance and undersampling. This design method can be extended to include other objective functions for different behaviors. PMID- 14559341 TI - Assessment of ESR-CT imaging by comparison with autoradiography for the distribution of a blood-brain-barrier permeable spin probe, MC-PROXYL, to rodent brain. AB - Blood-brain-barrier (BBB)-permeable, 3-methoxycarbonyl-2,2,5,5 tetramethylpyrrolidine-1-yloxy (MC-PROXYL) and BBB-impermeable carbamoyl-PROXYL were used to assess the ESR imaging technique by comparing with autoradiography. For this purpose, spin probes, 14C-labeled at their five-membered ring, [14C]MC PROXYL and [14C]carbamoyl-PROXYL, were newly synthesized. These probes were i.p. or i.v. injected into rats and autoradiograms were recorded. The autoradiograms of rat head showed that [14C]MC-PROXYL distributed well in the brain compared to [14C]carbamoyl-PROXYL. In vivo ESR spectra and two-dimensional ESR images of isolated rat brain treated with MC- or carbamoyl-PROXYL also indicated the extensive distribution of MC-PROXYL but not carbamoyl-PROXYL in the rat brain. The three-dimensional ESR images of the head of rats and mice were consistent with the fact that MC-PROXYL but not carbamoyl-PROXYL is incorporated into the brain. The ESR-CT images were better for mice than rats. However, the quality of the ESR-CT images was still not satisfactory. Although the resolution and sensitivity of the ESR-CT images were worse than those of the autoradiographic images, the former technique has unique features and advantages; e.g., functional, noninvasive and three-dimensional detection. PMID- 14559342 TI - MRI of the lung gas-space at very low-field using hyperpolarized noble gases. AB - In hyperpolarized (HP) noble-gas magnetic resonance imaging, large nuclear spin polarizations, about 100,000 times that ordinarily obtainable at thermal equilibrium, are created in 3He and 129Xe. The enhanced signal that results can be employed in high-resolution MRI studies of void spaces such as in the lungs. In HP gas MRI the signal-to-noise ratio (SNR) depends only weakly on the static magnetic field (B(0)), making very low-field (VLF) MRI possible; indeed, it is possible to contemplate portable MRI using light-weight solenoids or permanent magnets. This article reports the first in vivo VLF MR images of the lungs in humans and in rats, obtained at a field of only 15 millitesla (150 Gauss). PMID- 14559343 TI - Single-voxel proton MR spectroscopy in toluene abuse. AB - Inhalation of toluene, which is an organic solvent, causes toxic encephalopathy characterized by cognitive impairment, cerebellar and extra-pyramidal symptoms. We studied cranial MR images and single-voxel MR spectroscopy of 22 toluene abusers and age-matched control subjects. The mean age of the abusers and mean duration of abuse were 18,1 years and 47 months, respectively. We got three MR spectra from the centrum semiovale, cerebellum and thalamus by using STEAM sequence with a TE value of 30 ms. N-acetyl aspartate (NAA)/Creatine (Cr), Choline (Cho)/Cr, myo-inositol (mI)/Cr peak integral ratios were calculated. NAA/Cr in the cerebellum and centrum semiovale of the abusers were significantly lower than those of the control subjects. mI/Cr in centrum semiovale and cerebellum were higher in toluene abusers. No significant difference was found in the metabolite ratios of the thalami. The association of NAA/Cr and mI/Cr ratios in cerebellum and centrum semiovale with the duration of abuse was significant. Normal level of NAA in thalamus, which was a neuron rich gray matter structure, might imply that toluene inhalation did not cause direct neuronal injury. Selective reduction of NAA and increased level of mI in white matter supported the theory of that axonopathy and gliosis were the main mechanisms of pathophysiology in chronic toluene encepholopathy. Insignificance of elevation of Cho/Cr ratios demonstrated that toluene inhalation did not cause active demyelination. PMID- 14559344 TI - Molality as a unit of measure for expressing 1H MRS brain metabolite concentrations in vivo. AB - Absolute concentrations of cerebral metabolite in in vivo 1H magnetic resonance spectroscopy studies (1H-MRS) are widely reported in molar units as moles per liter of tissue, or in molal units as moles per kilogram of tissue. Such measurements require external referencing or assumptions as to local water content. To reduce the scan time, avoid assumptions that may be invalid under specific pathologies, and provide a universally accessible referencing procedure, we suggest that metabolite concentrations from 1H-MRS measurements in vivo be reported in molal units as moles per kilogram of tissue water. Using internal water referencing, a two-compartment water model, a simulated brain spectrum for peak identification, and a spectroscopic bi-exponential spin-spin relaxation segmentation technique, we measured the absolute concentrations for the four common 1H brain metabolites: choline (Cho), myo-inositol (mIno), phosphocreatine + creatine (Cr), and N-acetyl-aspartate (NAA), in the hippocampal region (n = 26) and along the Sylvian fissure (n = 61) of 35 healthy adults. A stimulated echo localization method (20 ms echo time, 10 ms mixing time, 4 s repetition time) yielded metabolite concentrations, uncorrected for metabolite relaxation or contributions from macromolecule resonances, that were expectantly higher than with molar literature values. Along the Sylvian fissure the average concentrations (coefficient of variation (CV)) in mmoles/kg of tissue water were 17.6 (12%) for NAA, 14.2 (9%) for Cr, 3.6 (13%) for Cho, and 13.2 (15%) for mIno. Respective values for the hippocampal region were 15.7 (20%), 14.7 (16%), 4.6 (19%), and 17.7 (26%). The concentrations of the two regions were significantly different (p 50%). At necropsy the mesh was excised en bloc with the anterior abdominal wall for histological evaluation of mesothelial layer growth. RESULTS: The mean adhesion score for the polypropylene mesh was significantly greater (P < 0.05) than Dual Mesh at 9 weeks and 16 weeks and modified Dual Mesh at 7 days, 9 weeks, and 16 weeks. Fifty-five percent (n = 11) of the polypropylene mesh had adhesions to small intestine or omentum at necropsy compared to 30% (n = 6) of the Dual Mesh and 20% (n = 4) of the modified Dual Mesh. There was a significantly greater percentage (P < 0.003) of ePTFE mesh mesothelialized at explant (modified Dual Mesh 44.2%; Dual Mesh 55.8%) compared to the polypropylene mesh (12.9%). CONCLUSIONS: Serial microlaparoscopic evaluation of intraperitoneally implanted polypropylene mesh and ePTFE mesh in a rabbit model revealed a progression of adhesions to polypropylene mesh over a 16 week period. The pore size of mesh is critical in the development and maintenance of abdominal adhesions and tissue ingrowth. The macroporous polypropylene mesh promoted adhesion formation, while the microporous nature of the visceral side of the ePTFE served as a barrier to adhesions. PMID- 14559438 TI - Mechanical properties of acellular peripheral nerve. AB - BACKGROUND: Acellular nerve has been used in experimental models as a peripheral nerve substitute. Our objective was to determine the difference in tensile strength between fresh and chemically treated acellularized peripheral nerve. MATERIALS AND METHODS: F344 rat sciatic nerves were either fresh or acellularized and tested either whole (Part A) or transected and repaired (Part B). For all constructs, the mean ultimate stress, mean ultimate strain, Young's modulus, and total mechanical work to fracture were calculated. The average ultimate strains for Groups A-1 and A-2 were 0.480 +/- 0.117 and 0.810 +/- 0.114, respectively. The Young's moduli in Groups A-1 and A-2 were 576 +/- 160 and 580 +/- 150 kPa, respectively. In Groups A-1 and A-2, the normalized work to failure was 0.35 +/- 0.14 and 1.11 +/- 0.38 N. The specimens in Group B-1 withstood an average ultimate stress of 780 +/- 280 kPa. The specimens in Group B-2 withstood an average ultimate stress of 405 +/- 20 kPa. RESULTS: The average ultimate strains for Groups B-1 and B-2 were 0.319 +/- 0.087 and 0.266 +/- 0.019, respectively. The Young's moduli in Groups B-1 and B-2 were 4,030 +/- 1360 and 2,290 +/- 280 kPa, respectively. The normalized work to failure in Groups B-1 and B-2 was calculated as 0.22 +/- 0.04 and 0.11 +/- 0.02 N. CONCLUSIONS: Although adequately robust for reconstructive procedures, the acellular peripheral nerve had decreased tensile strength compared with fresh nerve either when tested whole or when transected and repaired. PMID- 14559439 TI - Safety of radiofrequency ablation of myocardium and lung adjacent to the heart: an animal study. AB - BACKGROUND: The purpose of this work was to assess the feasibility and safety of applying radiofrequency to lung parenchyma adjacent to the heart and to evaluate the clinical impact and the histopathological damage to the heart caused by radiofrequency electrodes penetrating the pericardium and the myocardium. MATERIALS AND METHODS: Three sheep underwent pulmonary radiofrequency ablation adjacent to the heart at 460 kHz with target temperatures of 70 degrees C for 10 min at a maximum deployment of 4 cm. The animals were euthanized at the end of the procedure and the lungs and hearts were removed for gross pathological examination. Histopathology on selected hematoxylin & eosin-stained specimens was performed. RESULTS: Circulating blood had a "heat-sink" effect with tissue temperature at the electrode tips adjacent to the heart not exceeding 47 degrees C. Ventricular tachycardia and occasional extrasystoles were the only adverse reactions noted, even when electrodes penetrated the myocardium heart rhythm returned to sinus upon discontinuation of the radiofrequency energy delivery. CONCLUSIONS: Pulmonary radiofrequency ablation next to the heart is a relatively safe procedure. Major concern is that incomplete tumor ablation adjacent to the heart is probable as a result of the cooling effect of the circulating blood. PMID- 14559440 TI - Sustained function in atrophying liver tissue after portal branch ligation in the rat. AB - BACKGROUND: Preoperative segmental portal vein occlusion has become a common method to prevent liver failure after extended hepatic resection. To date, it is not elucidated whether atrophy by portal deprivation with concomitant contralateral regeneration leads to impaired liver function. We addressed this question by examining the expression of liver function proteins related to glucose homeostasis and acute-phase response in a corresponding animal model. MATERIALS AND METHODS: Male Wistar rats were subjected to either portal branch ligation (PBL), partial hepatectomy (PH), or sham operation (SO). The mRNA expression and chronological distribution of glucose-6-phosphatase (G6P), glucagon receptor (GR), glceraldehyd-3-phosphate-dehydrogenase (GAPDH), albumin, fibronectin, and C1-esterase-inhibitor (C1-Inh) genes were examined by Northern blot hybridizations. Determinations of serum-glucose and glycogen staining by periodic acid and Schiff were performed to analyze changes in glucose mobilization and storage. RESULTS: In regenerating liver tissue after PH and PBL, we detected a selective reduction of transcripts encoding G6P during the prereplicative period 6 and 12 h after surgery and a contemporary drop in serum glucose levels. This impairment proved to be more distinct after PH than after PBL. Compared with the residual liver after PH, the level of glycogen disappearance was lower after PBL in the regenerating lobe. In the portal deprived liver tissue, the expression of genes coding for G6P, GR, GAPDH, albumin, fibronectin, and C1-Inh was not altered compared with the SO group. CONCLUSIONS: Overall, portal-deprived liver tissue undergoing atrophy retains its liver-specific differentiation and function and helps to maintain homeostasis during the fast regeneration of the non-occluded liver lobe. PMID- 14559441 TI - A valid method of laparoscopic simulation training and competence assessment. AB - BACKGROUND: The purpose of our study was to evaluate the construct validity of laparoscopic technical performance measures and the face validity of three laparoscopic simulations. MATERIALS AND METHODS: Subjects (N = 27) of varying levels of surgical experience performed three laparoscopic simulations, representing appendectomy (LA), cholecystectomy (LC), and inguinal hemiorrhaphy (LH). Five laparoscopic surgeons, blinded to the identity of the subjects, rated the subjects on procedural competence on a binary scale and in four skills categories on a 5-point scale: clinical judgment, dexterity, serial/simultaneous complexity, and spatial orientation. Using a task-specific checklist, non clinical staff assessed the technical errors. The level of surgical experience was correlated with the ratings, the technical errors, and the time for each procedure. Subject responses to a survey regarding the utility of the inanimate models were evaluated. RESULTS: Years of experience directly correlated with the skills ratings (all P < 0.001) and with the competence ratings across the three procedures (P < 0.01). Experience inversely correlated with the time for each procedure (P < 0.01) and the technical error total across the three models (P < 0.05). Nearly all subjects agreed that the corresponding procedures were well represented by the simulations (LA 96%, LC 96%, LH 100%). CONCLUSION: The laparoscopic simulations demonstrated both face and construct validity. Regardless of the level of surgical experience, the subjects found the models to be suitable representations of actual laparoscopic procedures. Task speed improved with surgical experience. More importantly, the quality of performance increased with experience, as shown by the improvement in the skills assessments by expert laparoscopic surgeons. PMID- 14559442 TI - Chemokine response of pulmonary artery endothelial cells to hypoxia and reoxygenation. AB - BACKGROUND: Chemokines are inflammatory mediators that activate and recruit specific leukocyte subpopulations. We have recently shown a role for certain chemokines in a warm in situ rat model of lung ischemia reperfusion injury. After hypoxic stress, rat pulmonary artery endothelial cells (RPAECS) potentiate and direct neutrophil sequestration, and, therefore, contribute to the development of tissue injury. The present studies were performed to determine whether RPAECS subjected to in vitro hypoxia and reoxygenation (H&R) secrete chemokines, and, if so, to define the regulatory mechanisms involved. MATERIALS AND METHODS: RPAECS were isolated from 21-day-old Long-Evans rats and were rendered hypoxic (pO(2) 0.5%) for 2 hours and reoxygenated for up to 6 hours. Secreted chemokine content was quantified using sandwich enzyme-linked immunosorbent assay techniques. Mechanistic studies assessed chemokine messenger ribonucleic acid (mRNA) expression by Northern blot, as well as the nuclear translocation of proinflammatory transcription factors nuclear factor kappa beta (NFkappaB), early growth response (EGR), and activator protein-1 (AP-1) by electromobility shift assays. Supershift analysis for EGR-1 was also performed. RESULTS: RPAECS showed a marked increase in the secretion of the chemokines cytokine induced neutrophil chemoattractant and monocyte chemoattractant protein-1 in response to H&R, which was dependent on de novo mRNA transcription and protein translation. Furthermore, in vitro H&R induced the nuclear translocation of the proinflammatory transcription factors NFkappaB and EGR-1 early during reoxygenation. CONCLUSIONS: RPAECS secrete significant amounts of cytokine induced neutrophil chemoattractant and monocyte chemoattractant protein-1 in response to in vitro H&R. The secretion of both chemokines is dependant on de novo mRNA transcription and protein translation, and may be regulated by NFkappaB and EGR-1 activation. PMID- 14559443 TI - Sustained bone mineral density changes after burn injury. AB - BACKGROUND: Body-composition changes have been observed after burn injury. In particular, several studies have shown that bone mineral density (BMD) in burn patients is decreased when compared to the normal population. Little is known about the frequency, severity, or duration of these changes. The purpose of this study was to describe body-composition changes over time after burn injury. MATERIALS AND METHODS: Twenty-nine burn patients participated in this study. Portable dual-energy X-ray absorptiometry (pDEXA) measuring forearm BMD, fat, and lean mass was obtained as soon as possible after admission and repeated bi-weekly until discharge and, when possible, for 2 years post-injury. The scan showing the greatest change in BMD, fat, or lean mass was compared to the baseline scan for each individual. RESULTS: Although lean and fat mass did not change significantly after injury, BMD decreased significantly. The greatest change in BMD did not occur during the acute burn hospitalization, but rather 131 days after burn injury. Changes in post-burn BMD inversely correlated with % total body surface area (TBSA) and % 3rd-degree TBSA. The magnitude of change was similar between adults and children. CONCLUSIONS: These results confirm earlier studies, suggesting that BMD can be negatively altered post-injury, with the greatest changes occurring after patients are discharged from the hospital. Although the clinical significance of these changes is not known, this study supports the need for long-term musculoskeletal assessments in burn patients and for further research to elucidate the mechanisms of burn-induced body-composition changes. PMID- 14559444 TI - Vascular endothelial growth factor inhibits mitogen-induced vascular smooth muscle cell proliferation. AB - INTRODUCTION: Delivery of vascular endothelial growth factor (VEGF) protein or gene transfer has been shown to accelerate re-endothelialization and attenuate neointimal hyperplasia in various arterial injury models, including balloon injury, stent implantation, and vein grafts. In addition to stimulating re endothelialization, we hypothesize that VEGF has further vascular protective functions to prevent neointimal hyperplasia by directly inhibiting mitogen induced proliferation of vascular smooth muscle cells (VSMCs) via the mitogen activated protein kinase pathway. MATERIALS AND METHODS: Human aortic VSMCs were seeded and serum starved for 24 h. The cells were then stimulated with a mitogen, recombinant human platelet derived growth factor at 20 ng/mL together with 0, 10, 20, 30, 40, 50 ng/mL recombinant human VEGF. A proliferation assay was used to quantitate bromodeoxyuridine uptake into newly synthesized DNA. Western immunoassay was used to quantify extracellular signal-regulated kinase (ERK) 2 protein and phosphorylation of retinoblastoma and ERK 1/2 protein. RESULTS: VEGF inhibited bromodeoxyuridine incorporation into mitogen-induced VSMC in a dose dependent manner, reaching statistical significance at concentrations of 30 (P < 0.05), 40 (P < 0.05), and 50 ng/mL (P < 0.01). Densitometry of western immunoblots revealed an inhibition of phosphorylation of retinoblastoma at VEGF concentrations of 40 and 50 ng/mL and ERK 1/2 phosphorylation at concentrations of 30, 40 and 50 ng/mL. CONCLUSION: In addition to stimulating re endothelialization, VEGF appears to have a vascular protective function by directly inhibiting VSMC proliferation. This effect occurs in the absence of endothelial cells and via the mitogen-activated protein kinase pathway. VEGF may serve as an important modulator of mitogen-induced VSMC proliferation after vascular injury. PMID- 14559446 TI - Investigation of muscle pH as an indicator of liver pH and injury from hemorrhagic shock. AB - BACKGROUND: During hemorrhagic shock blood flow to vital organs is maintained by the diversion of blood from both the splanchnic organs and skeletal muscle. In this swine study, we tested the hypotheses that (1). liver and muscle pH are correlated during both shock and resuscitation and (2). muscle pH during shock is an indicator of potential liver injury after resuscitation. MATERIALS AND METHODS: Hemorrhagic shock was induced over 15 min to lower systolic blood pressure to 40 mm Hg and was maintained for 60 (n = 5) or 90 (n = 5) min. Resuscitation was achieved with shed blood and warm saline to maintain mean pressure >60 mm Hg for 120 min. Liver and muscle pH were measured with microelectrodes throughout the entire shock and resuscitation periods, along with hepatic venous oxygen saturation. Arterial lactate and aspartate aminotransferase were measured at baseline, end of shock, and resuscitation. Correlation between muscle and liver pH was determined. The ability of muscle pH to predict liver injury (40% increase in arterial aspartate aminotransferase) was compared with other predictors: liver pH, arterial lactate, and tonometric-arterial PCO(2) gap. RESULTS: pH values and rates of change were similar in both muscle and liver tissue. Liver pH was well correlated with muscle pH during both shock and resuscitation, R(2) = 0.84. Muscle pH predicts potential liver injury with the same sensitivity as blood lactate in this swine shock model. CONCLUSIONS: Minimally invasive measurement of muscle pH warrants further study as a method to assess splanchnic hypoperfusion and resultant injury. PMID- 14559445 TI - HL-1 myocytes exhibit PKC and K(ATP) channel-dependent delta opioid preconditioning. AB - BACKGROUND: Opioid preconditioning protects the myocardium against ischemia/reperfusion (IR) injury. By enhancing cardiomyocyte viability, opioids can enhance cardiac function and recovery from IR injury during acute cardiac care. The myocyte model HL-1 is an immortalized, mouse atrial cell line that expresses functional delta-opioid receptors. The HL-1 myocyte may be useful for IR injury research exploring opioid cardioprotection. MATERIALS AND METHODS: In study I, microplates of HL-1 were subjected to 10 min pre-treatment with either basal media, delta-opioid agonist DADLE(10uM), or DADLE(10uM) + delta-antagonist naltrindole (10uM). Study II treatment groups included PKC inhibitor chelerythrine (2uM), K(ATP) channel closer glybenclamide (100uM), or mitochondrial K(ATP) channel opener diazoxide (100uM) administered in various combinations followed by DADLE (10uM) or control. Microplates were subjected to normal oxygen/substrate conditions or ischemic (<1% 0(2)) and substrate deficient (10 uM 2-Deoxyglucose versus 10 mM glucose) conditions, then reperfused with normal oxygen and glucose-containing media. Microplate supernatants were subjected to lactate dehydrogenase (LDH) assay. RESULTS: Compared to untreated control, the LDH assay showed significant reduction in opioid-only pretreated groups at all time points. These effects were attenuated with delta-opioid antagonist co-administration. Co-administration of non-selective K(ATP) channel closer glybenclamide and DADLE abolished DADLE cytoprotection, while selective mitochondrial K(ATP) opener diazoxide mimicked DADLE cytoprotection Co administration of chelerythrine and DADLE significantly reduced chelerythrine cytotoxicity. CONCLUSION: Delta-opioid preconditioning of HL-1 myocytes significantly decreased necrosis from in vitro simulated ischemia/reperfusion as measured by LDH release; this effect was reversed by delta-antagonist naltrindole. Cytoprotection was PKC and K(ATP) channel-dependent. HL-1 myocytes exhibit opioid-induced cytoprotection from IR injury, and present a novel model of pharmacologic preconditioning. PMID- 14559447 TI - Effect of bladder outlet obstruction on detrusor smooth muscle cell: an in vitro study. AB - BACKGROUND: Although relieving obstruction is generally curative on bladder outlet obstruction (BOO), bladder dysfunction persists in some patients. Repetitive stretch and relaxation applied to cultured bladder smooth muscle (SM) cells in vitro have been used to mimic increases in urodynamic load experienced by the detrusor muscle under conditions of BOO. We first clarified the relationship between phenotype transformation and biomechanical properties of detrusor smooth muscle cell (DSMC) subjected to the cyclic mechanical stretch. MATERIALS AND METHODS: Cultured rat DSMC were grown on collagen-coated silicone membranes and subjected to continuous cycles of stretch-relaxation. All experiments were performed on cells between passages 2 and 4. Each cycle consists of 5 seconds of stretch and 5 seconds of relaxation. The computer controlled vacuum induced 10% (1), 20% (2), and 30% (3) maximum elongation of the plate membrane at different designed pressures. The deoxyribonucleic acid synthesis rate was assessed by performing tritiated thymidine incorporation assay. The expression of SM-alpha-actin and proliferation of DSMC were analyzed by immunofluorescent assay and flow cytometry. The image analysis and micropipet aspiration systems were used to investigate the single cell contraction and viscoelasticity. Using the 3-element standard linear solid model, the elastic modulus K(1), K(2), and viscoelastic coefficient mu were determined, which show the passive deformation ability of detrusor cells. RESULTS: As the basic structural changes to mechanical stretch, DSMC undergo phenotypic modulation from their normal contractile phenotype to a "synthetic" phenotype: the DSMC become more proliferative and the actin less organized along the cell's long axis. The cell proliferation index of control and stretched group (10%, 20%, 30% elongation) are 0.24, 0.43, 0.58, and 0.65, respectively. The actin filaments in unstimulated cells were evident and orientated along the major axis of the cell. After mechanical stretch, the well-spread filaments changed their orientation. The function, such as contraction, and viscoelasticity of a single DSMC subjected to stretch both decreased significantly compared with control. The maximum contractile velocity and maximum cell length shortening rate of group 3 (30% elongation) showed significant decreases compared with unstretched control (P < 0.01). K(1) and K(2) were decreased with the increase of mechanical overload. However, there was no statistic difference between groups 2 and 3. CONCLUSIONS: Functional abnormalitie of BOO have the structural basis: phenotype transformation (i.e., remodeling) of the detrusor cells. Cyclic stretch and relaxation applied to DSMC in vitro can be used to model increases in urodynamic load experienced by the bladder detrusor muscle under conditions of BOO. Phenotype transformation is the structural basis of functional changes of DSMC subjected to periodic overload mechanical stretch. PMID- 14559676 TI - Arsenic trioxide-induced apoptosis of human malignant lymphoma cell lines and its mechanisms. AB - OBJECTIVE: To investigate the responses of human Burkitt lymphoma cells to arsenic trioxide (As2O3) and the possible mechanisms. METHODS: Epstein-Barr virus (EBV)-positive human B-lymphoma Raji cell line and EBV-negative human B-lymphoma BJAB cell line were used as in vitro models to assess the cell apoptosis by morphology and DNA agarose gel electrophoresis. Protein expression was analyzed using Western blotting. RESULTS: After 24-hour treatment with the 2, 5 and 10 micromol/L As2O3, the concentrations of As2O3 achievable in vivo, cell apoptosis was induced in human Burkitt lymphoma BJAB cells at the rates of 47.6%+/-4.8% (Mean+/-SD, n=3), 66.4%+/-5.1%, 87.0%+/-7.3% and at 35.5%+/-3.8%, 51.5%+/-6.2%, 62.2%+/-7.9% respectively in Raji cells, corresponding to the concentration of As2O3. EBV-infected Raji cell line was less sensitive to As2O3 than EBV-negative BJAB cell line (P<0.05). As2O3-induced apoptosis was accompanied by down regulation of Bcl-xL protein expression and activation of apoptosis protein caspase-3, as identified by Western blotting. CONCLUSION: As2O3 exerts apoptosis inducing effects on human Burkitt lymphoma cells through down-regulation of Bcl xL protein expression and activation of apoptosis protein caspase-3, and may serve as a candidate therapeutic agent against malignant lymphoma for both systemic and local therapies. PMID- 14559677 TI - Etiological analysis of the sequelae of radiotherapy for nasopharyngeal carcinoma: a follow-up study of 112 cases. AB - OBJECTIVE: To explore the etiology of the sequelae of radiotherapy for nasopharyngeal carcinoma (NPC) so as to find the possible means for reducing or preventing its occurrence. METHODS: A total of 112 pathologically confirmed patients with nasopharyngeal carcinoma, who had survived for 5 years following the radiotherapy, were included in this study. Sixty-four patients with the primary carcinoma in the nasopharyngeal region received radiotherapy with the radiation field covering the bilateral anterior ear regions, and in the other 48 patients, adjuvent exposure of the anterior nasal region was administered. The metastases in the cervical lymph nodes were exposed to tangential radiation by 40 Gy X-ray followed by approximately 20 Gy vertical electron beam exposure. RESULTS: Limited mouth-opening and dry mouth occurred mostly during the first 2 years after radiotherapy, and hearing loss in the first year. Neck fibrosis tended to increase with the time elapse after the therapy, and posterior cranial nerve damages showed no signs of time-related occurrence. It was found that the occurrence of neck fibrosis, dry mouth and the nerve injuries did not obviously correlate with the dosage of X-ray exposure in the anterior ear regions, while limited mouth-opening and the hearing loss increased with the radiation dosage. The sequelae did not arise from different radiation field selection as adopted in this study. CONCLUSION: The radiation dose should be controlled at around 70 Gy for NPC treatment, and for carcinoma remnant in the nasopharyngeal region, additional dose in the cavity would be appropriate. When the X-ray dose of 65 Gy in the neck region fails to result in satisfactory recession of lymph node metastasis, comprehensive treatment involving multiple modalities should be considered. PMID- 14559678 TI - [Establishment of animal models of endometriosis in rhesus monkeys]. AB - OBJECTIVE: To establish animal models of endometriosis in rhesus monkeys and study the invasion mechanism of endometriosis. METHODS: Five female healthy rhesus monkeys with regular menstrual cycles were selected, of whom 4 were used for endometriosis model establishment and 1 for control. During the period of days 8-15 of menstruation, or 3-5 days after the midcycle estrogen peak, the endometrium was implanted into the pelvic cavity other than the uterus in the experimental group, and in the control group, the greater omentum was implanted instead. Exploratory laparotomy or laparoscopy was performed during the period between days 8 and 15 of menstruation in the second and the fourth month respectively after the implantation for observing the occurrence of endometriosis. RESULTS: In the experimental group, the implanted endometrium survived in the receptor site in 3 monkeys, 2 of whom were found to have chocolate cyst after the operation. Laparoscopy was again performed in the 2 monkeys with endometriosis 4 months after the operation, showing pelvic adhesion and growth of ectopic endometrium similar to the changes as observed two months after the operation. In comparison with the control, deteriorated mental states and dietary changes, along with body weight losses, occurred in the 3 models of endometriosis, and 2 of them later died of intussusception. CONCLUSIONS: Animal models of endometriosis can be established successfully in rhesus monkeys on the basis of the implantation theory, which, however, is influenced by individual differences between the animals. Heredity is an important factor in pathogenesis of endometriosis, and ectopic growth of the endometrium may severely impact the body function. PMID- 14559680 TI - [Apoptosis of keloid-derived fibroblasts induced by Fas gene transfection]. AB - OBJECTIVE: To observe the effect of Fas gene transfection on the apoptosis of keloid-derived fibroblasts. METHODS: The full-length cDNA of Fas was inserted into the multicloning site of the expression vector pcDNA-3.1 by molecular cloning technique, and the recombinant plasmid was transfected into the keloid derived fibroblasts via lipofectin. Fas monoclonal antibody (mAb) was used to induce the apoptosis of the cells, which was evaluated with HE staining, DNA agarose gel electrophoresis and flow cytometry. RESULTS: The expression plasmid pcDNA-3.1-Fas was constructed successfully. Fas mAb induced apoptosis of these fibroblasts transfected with Fas gene, with the typical features of fibroblast apoptosis observed in the treated cells (e.g. typical apoptotic cell nuclei, DNA ladder and high apoptosis rate as determined by flow cytometer), but not in the control cells. CONCLUSION: Blockage of upstream apoptosis pathway in keloid derived fibroblasts is due to nonfunction of Fas protein, and mutation of Fas gene may be the pathogenesis of keloids. PMID- 14559679 TI - [Analysis of short tandem repeat polymorphism in a female patient with Duchenne muscular dystrophy]. AB - OBJECTIVE: To understand the mechanism of Duchenne/Becker muscular dystrophy (DMD) in a female patient. METHODS: A multiplex PCR (mPCR) protocol was applied to detect the dystrophin gene of the female DMD patient and her family members, whose haplotypes were analyzed in light of short tandem repeat polymorphism (STR) of five microsatellite markers (located in 5' terminus and introns 44, 45, 49, and 50). RESULTS: No deletion was detected in the affected female patient and her affected son. Examination of the female DMD patient's STR haplotypes identified in the female patient and her affected son the same haplotype inherited from her unaffected mother, who was a likely germinal mosaicism. The female patient has different haplotype from her affected son and her mother as a deletion was identified in the intron 45 of the affected son, and the female patient and her mother were probably heterozygous for this deletion. CONCLUSION: STR haplotype may not be responsible for the pathogenesis of DMD in the female patient and her affected son. PMID- 14559681 TI - [Cloning and expression of hepatitis C core protein gene]. AB - OBJECTIVE: To express hepatitis C virus (HCV) core protein gene fragment in E. coli. METHODS: A fragment of HCV core gene sequence 357 bp in length was amplified by PCR, digested with EcoRI+Hind III and inserted to the plasmid vector pET-32a to construct recombinant HCVc/pET-32a plasmid, which was transformed into E.coli BL-21 and induced by IPTG for its expression. The expressed proteins obtained were identified by SDS-PAGE and Western blotting. RESULTS: The core sequence of HCV was amplified and after IPTG induction, a fusion protein of 32,000 was resulted exhibiting specific reaction with HCV-positive serum and high antigenicity. CONCLUSION: It is possible to efficiently express HCV core protein in E.coli. PMID- 14559682 TI - [Effect of bone morphogenetic protein microspheres on biological behavior of rabbit bone marrow stem cells]. AB - OBJECTIVE: To evaluate the effect of bone morphogenetic protein (BMP) on the biological behavior of bone marrow stem cells (BMSCs) of rabbits. METHODS: BMP was either enwrapped or not in the microspheres made of chitosan and sodium alginate, and the biocompatibilities of the composites were examined by means of cell culture. The BMSCs were cultured with the two kinds of microspheres respectively, and the cell extension rate, proliferation, alkaline phosphatase activity and Coomassie blue staining of the cells were assayed. RESULTS: Inhibition of BMSC proliferation did not occur in response to in vitro culture with the microspheres, but alkaline phosphatase activity and D(lambda) values of Coomassie blue staining increased significantly in the cells cultured with BMP microspheres. CONCLUSION: BMP can increase the osteogenic capacity of BMSCs in vitro with the microspheres made of chitosan and sodium alginate as the carrier. PMID- 14559683 TI - [Association of receptor-mediated endocytosis and autophagy with apoptosis]. AB - OBJECTIVE: To evaluate the relationship between receptor-mediated endocytosis, autophagosome formation and apoptosis by observing the morphological changes of mouse peritoneal macrophages in the course of receptor-mediated endocytotic activities, autophagy and apoptosis. METHODS: Mouse peritoneal macrophages were incubated with horseradish peroxidase-labeled concanavalin A (ConA-HRP), and morphological examinations were performed at different time points after the incubation. RESULTS: After the incubation of the macrophages with ConA-HRP, ConA HRP was observed to enter the vesicles by way of receptor-mediated endocytosis. Three kinds of endosomes were observed, namely vesicles, tubes and double membrane sheets. The double-membrane sheets enveloped a portion of the cytoplasm and organelles, thus giving rise to vesicles, or the autophagosomes, which later fused with lysosome, followed by the apoptosis of the macrophages. CONCLUSION: Receptor-mediated endocytosis of ConA-HRP is associated with autophagy and apoptosis. PMID- 14559684 TI - [Modified type I thyroplasty for treating unilateral vocal cord paralysis]. AB - OBJECTIVE: To explore a new surgical approach for treating unilateral vocal cord paralysis. METHODS: Five cases of unilateral vocal cord paralysis due to various causes were treated with modified type I thyroplasty. Laryngostroboscopy and electroglottography were performed before and after the operations. RESULTS: The vocal cords were shifted very close to the median line in all the 5 cases postoperatively as shown by laryngostroboscopy and electroglottography. Complete closure of the glottis was achieved. The patients had almost normal results of electroglottography, and the vocal sound nearly recovered normal or was significantly improved. CONCLUSION: Modified type I thyroplasty is effective, safe and easy in treating unilateral vocal cord paralysis. PMID- 14559686 TI - [Effect of echo-contrast agent on rat vascular smooth muscle cell proliferation during ultrasound exposure]. AB - OBJECTIVE: To investigate the effect of echo-contrast agent on rat vascular smooth muscle cell (VSMC) proliferation in the presence of ultrasound exposure. METHODS: The VSMCs of rats were cultured in 6-well plates with or without the echo-contrast agent and stained with trypan blue at 3, 24, and 48 h respectively after ultrasound exposure for 1 min at 2 MHz, 0.25 mechanical index (MI). At each time point as indicated after the exposure, the cells in parallel wells were trypsinized for cell counting using Coulter counter. RESULTS: No cytotoxicity was found in trypan blue staining. Compared with the control cells, the proliferation of VSMCs was inhibited by echo-contrast agent and adjunctive ultrasound within 24 h, but another 24 h later, the effect failed to be observed. CONCLUSIONS: The proliferation of VSMC was inhibited by with low mechanical index cavitation activity of contrast agents. PMID- 14559685 TI - [Mechanism of quercetin as an antidiarrheal agent]. AB - OBJECTIVE: To investigate the antidiarrheal mechanism of quercetin extracted from Psidium guajava L. METHODS: The effects of quercetin on the contractility of guinea pig ileum in vitro and on the peristaltic motion of mouse small intestine in vivo were observed, and the peak value of contractility and peristaltic distance recorded. The inhibitory effect of quercetin on the permeability of the abdominal capillaries was also observed. RESULTS: Quercetin inhibited the contraction of guinea pig ileum in vitro and the peristaltic motion of mouse small intestine, and reduced the permeability of abdominal capillaries. CONCLUSION: Quercetin can inhibit the intestinal movement and reduce capillary permeability in the abdominal cavity, which may be the antidiarrheal mechanism of Psidium guajava L extract. PMID- 14559687 TI - [Application of suppression subtractive hybridization technique in the investigation of heat adaptation mechanism]. AB - OBJECTIVE: To understand whether there are genes differentially expressed in association with heat adaptation in resting heat-adapted rats at normal temperature, and to evaluate the feasibility of investigating heat adaptation mechanism in light of gene differential expression. METHODS: The mRNAs were extracted from rat liver tissues and reverse transcripted into double-strand cDNAs. Suppression subtractive hybridization (SSH) was applied to compare the gene expression profiles between heat adaptation group and normal temperature control group, and cDNA subtraction libraries of the differentially expressed genes were constructed. PCR-selective differential screening technique was used to identify the differentially expressed genes. RESULTS: Six cDNA fragments were identified in the preliminary experiment to be differentially expressed. CONCLUSIONS: There are genes differentially expressed in the liver of resting heat-adapted rats in association with heat adaptation at normal temperature, and these genes may be involved in maintaining heat adaptation. This study also indicates the feasibility of investigating heat adaptation mechanism by isolating differentially expressed genes associated with heat adaptation. PMID- 14559689 TI - [Immunophenotyping of 106 adult patients with acute leukemia by flow cytometry]. AB - OBJECTIVE: To study the immunophenotyping of adult patients with acute leukemia and its association with the prognosis. METHODS: Immunophenotyping was performed in 106 adult patients with acute leukemia by three-color flow cytometry analysis using CD34/SSC gating. RESULTS: The antigens expressed in 71 patients with acute myeloid leukemia (AML) were mainly CD13, CD33, HLA-DR, CD34 and CD117. Lymphoid antigen expression was identified in 23.9% adult AML patients and CD56 antigen expression in 15.5% of the AML patients. In 29 patients with acute lymphoblastic leukemia (ALL), the expressed antigens were mainly HLA-DR, CD10, CD19, CD34 and CD7, and 34.5% of these patients were found to be positive for myeloid antigen expression. Complete remission (CR) rate in AML patients with lymphoid antigen expression after chemotherapy was lower than that in AML patients without lymphoid antigen expression (52.9% vs 77.8%, P<0.05), and no significant impact was noted of myeloid antigen expression in ALL on the CR of the patients (70.0% vs 94.7%, P>0.05). The CR rate in AML with CD56 antigen expression was lower than that in AML without CD56 expression (36.4% vs 78.3%, P<0.025), and the CR rate in CD34+ AML was lower than that in CD34- AML (56.0% vs 80.4%, P<0.05). CONCLUSIONS: Gating of CD45/SSC can eliminate the interference of normal cells to render more reliable immunophenotyping results. The expressions of CD56+, CD34+, lymphoid antigen in adult AML patients, who have lower CR rate, often signify poor prognosis. PMID- 14559688 TI - [Detection of core-binding factor in acute leukemia with interphase fluorescence in situ hybridization]. AB - OBJECTIVE: To detect the chromosomal abnormalities involving the core-binding factor (CBF) in acute leukemia at initial diagnosis with interphase in situ hybridization (I-FISH) technique, and monitor the minimal residual disease (MRD) after treatment with I-FISH. This study also aim to compare the sensitivity of I FISH at initial diagnosis with that of conventional G-banding analysis. METHODS: Based on the diagnosis of bone marrow morphology, 15 acute leukemia patients were examined with conventional G-banding and I-FISH techniques. Seven of these patients were monitored for MRD with I-FISH after treatment. RESULTS: On the basis of the false-positive rate acquired from normal subjects, the normal cutoff values of the 3 probes including AML1/ETO translocation probe, MYH11 breakpoint region probe and ETV6/AML1 translocation probe were 4.13%, 1.95% and 2.12% respectively. With conventional G-banding analysis, 40% (6/15) patients were found with chromosomal abnormality involving CBF, including 5 of the 8 M2 patients with t (8;21) and 1 of the 2 M4EO patients with inv (16). No B-ALL cases were identified with t (12;21). With I-FISH, however, 80% (12/15) of the cases were found with genetic abnormality involving CBF, including all the 8 M2 cases with AML1/ETO fusion gene, both of the M4EO cases with CBFbeta/MYH11 and 2 of the 5 B-ALL cases with ETV6/AML1. In the 7 cases monitored for MRD level with I-FISH, 2 M2 cases and 1 B-ALL case were with positive results. CONCLUSIONS: I-FISH is more sensitive than conventional G-binding analysis in detecting the chromosomal abnormalities involving CBF in acute leukemia. At the time of initial diagnosis, combination of the two techniques may lead to more comprehensive and accurate results. PMID- 14559690 TI - [Study of adhesion-related molecule beta1-integrin and focal adhesion kinase in chronic myeloid leukemia]. AB - OBJECTIVE: To study the changes in the expressions of beta1-integrin receptor (CD29) and focal adhesion kinase (FAK) during the deterioration of chronic myeloid leukemia (CML), thereby explore the potential mechanism of interferon (IFN)-alpha in the treatment of CML through testing the quantitative changes of FAK. METHODS: Bone marrow mononuclear cells were tested for CD34, CD29 and FAK monoclonal antibody by flow cytometry in CML patients of the chronic phase and blast crisis and in normal subjects. In the presence or absence of IFN-alpha, the bone marrow mononuclear cells from CML patients of chronic phase and normal subjects were cultured for 48 h to determine the content of FAK in the cells with Western blotting. RESULTS: CD 29 expression on the surface of CD34+ cells scarcely differ between normal subjects and CML patients of chronic-phase, but the intracellular content of FAK was lower in the latter. The expression of beta1 integrin receptor on the surface of hematopoietic cells in CML patients with blast crisis was significantly higher than that in chronic-phase CML patients, but the intracellular FAK in the former patients was lower. No difference of FAK content was observed in normal mononuclear cells before and after IFN-alpha treatment, while the treatment increased FAK content in the mononuclear cells in chronic phase CML patients. CONCLUSION: The changes in the expressions of beta1 integrin receptor and FAK may play important roles during CML deterioration, and IFN-alpha may function to repair the defect in the beta1-integrin receptor pathway by restoring the cellular content of FAK. PMID- 14559691 TI - [Effect of autocrine vascular endothelial growth factor on the biological activity of acute myeloid leukemia cell line HL-60]. AB - OBJECTIVE: To explore the effect of autocrine vascular endothelial growth factor (VEGF) on the abnormal proliferation of HL-60 cells. METHODS: HL-60 cells were transfected via lipofictamine with the VEGF(165) cDNA sense vector (HL-60/ VEGF(165)) and with the PcDNA3.1(-)-vector (HL-60/neo) as the control. Reverse transcription-polymerase chain reaction (RT-PCR) was used for detecting VEGF mRNA in the transfected cells. VEGF concentrations in the cell culture supernatant were determined by enzyme-linked immunosorbent assay (ELISA), and cell proliferation was determined by MTT assay and colony-forming assay in vitro. Flow cytometric Annexin-V-FITC/ PI dual-labeling technique was employed to observe the effect of VEGF(165) cDNA transfection on the apoptosis of HL-60 cells. RESULTS: The levels of VEGF mRNA expression by HL-60/VEGF(165) cells were higher than HL 60/neo, with the mean VEGF quantity in the supernatant of HL-60/VEGF(165) cell culture being 399.07+/-12.45 ng/L, which was 2-fold higher than that in the supernatant of HL-60/neo cells (184.45+/-10.53 ng/L) (P<0.01). The growth rate of HL-60/VEGF(165) cells was significantly accelerated as compared with that of the control cells, and the colony formation capacity of the former cells also increased significantly (P<0.05), with the average colony number of 157.00+/ 17.00/500 cells vs 110.00+/-12.90/500 cells of HL-60/neo cells. Less apoptotic cells were identified among HL-60/VEGF(165) cells than in HL-60/neo cells in the same culture condition. CONCLUSION: Autocrine VEGF plays an important role in the proliferation and apoptosis of acute myeloid leukemia cells. PMID- 14559692 TI - [Modified FLAG regimen in the management of refractory acute myeloid leukemia]. AB - OBJECTIVE: To evaluate the therapeutic effect of modified FLAG regimen in the management of refractory acute myeloid leukemia (AML). METHODS: Sixteen patients with refractory AML were divided into two groups. In modified FLAG regimen group (n=10), the patients received fludarabine (Flu, 50 mg/d, VDx5 d) and Ara-c (200 mg/d, VDx5 or 7 d). The regimen for classic FLAG group (n=6) consisted of Flu (50 mg/d, VDx5d), Ara-C (500 or 1,000 mg/d, VDx5d) and G-CSF (300 microg/d, x5 d, subcutaneously injected 4-6 hours before chemotherapy). Each patient received subcutaneous G-CSF (300 microg/d) when the white blood cell count was lower than 1.0x10(9)/L till the condition was corrected. RESULTS: The total complete remission(CR) rate of the 16 patients was 50% (8/16). Seven patients in modified group achieved CR (70%) and only one of the classic group did (17%, P<0.05). Episodes of infections were lower in modified group than in the classic group (50% vs 83%). CONCLUSION: Modified FLAG regimen is more likely than classic FLAG regimen to achieve CR and reduce infections in patients with refractory AML. PMID- 14559693 TI - [Therapeutic effects of 3-dimensional conformal radiotherapy for small hepatocellular cancers and multi-factor analysis of the prognosis]. AB - OBJECTIVE: To evaluate the therapeutic effects of 3-dimensional conformal radiation therapy (3-DCRT) for small hepatocellular carcinoma (HCCs) and to analyze the prognostic factors. METHODS: Forty-two HCCs patients were treated with 3-DCRT and acute effects and survival rates were observed. Kaplan-Meier method was used for survival analysis, and Cox proportional hazard model adopted in multi-factor analysis. RESULTS: No evidence of local failure was found in 83.3% of the patients at 3 months after the radiotherapy. The overall 1-, 2-, and 3-year survival rates were 100.00%, 90.48%, 78.57% respectively. Multivariate analysis revealed that the gender, number of tumors, and tumor size were the independent and statistically significant factors affecting the patients' survival (P<0.05). CONCLUSIONS: 3-DCRT is effective and feasible for treating non operable HCCs, and the gender, number of tumors, and tumor size are the major factors affecting the survival of HCCs patients. PMID- 14559694 TI - [Effects of hypothermia on cerebral oxygen metabolism in patients with mitral valve replacement]. AB - OBJECTIVE: To observe the effects of mild to moderate hypothermia on cerebral oxygen metabolism in patients with mitral valve replacement. METHODS: Twenty patients undergoing mitral valve replacement were randomly divided in mild (30 ) and moderate (26 degrees Celsius) hypothermia groups. Under alpha stat, the oxygen content and the concentration of lactic acid in the radial artery and jugular venous bulb were monitored in patients undergoing mitral valve replacement at mild and moderate hypothermia respectively. The arterial-venous difference of oxygen content, oxygen uptake rate, and blood lactic acid levels in the cerebrum and total body were calculated. The effect of cardiopulmonary bypass (CPB) under the two hypothermia strategies on cerebral oxygen metabolism was analyzed. RESULTS: In the two groups, arterial-venous differences and oxygen uptake rates were both decreased after the commencement of cardiopulmonary bypass, and were rapidly elevated during rewarming. In mild hypothermia group, the arterial-jugular venous difference and oxygen uptake were higher than those in moderate hypothermia group (P<0.05) during CPB when the lowest temperature was reached. The concentration of lactic acid in the plasma was progressively increased in both groups during CPB. Arterial-venous difference in the oxygen content in both groups was still lower during CPB than before CPB(P<0.01). CONCLUSION: Mild and moderate hypothermia during CPB is sufficient to retain the balance of cerebral oxygen metabolism, and more intensive hypothermia may not ensure better cerebral protective effect. Cerebral oxygenation progressively increases after CPB under hypothermia, but whether imbalance of cerebral oxygen metabolism occurs needs further investigation. PMID- 14559695 TI - [Treatment of acute lung injury due to sea water aspiration in dogs by early massive bronchoalveolar lavage: an experimental study]. AB - OBJECTIVE: To investigate the therapeutic effects of massive bronchoalveolar lavage at early stage of acute lung injury induced by sea water aspiration. METHODS: Twelve mongrel dogs were subjected to sea water infusion of the entire lungs and were subsequently randomized into 2 groups. The dogs in group A received intravenous saline transfusion 24 h after injury, while those in group B underwent bronchoscopic lavage of injured lungs with 450 ml 0.45% saline 15 min after injury. Unassisted breathing was maintained in both groups, and the gas exchange function of the bilateral lungs and the levels of lactate dehydrogenase, alkaline phosphatase in the bronchoalveolar fluid were observed, with the hemodynamics indices monitored continuously. The lungs of the dogs were finally removed for morphological study under both light and electron microscopes, and the water content and wet- to dry-weight ratio of the lung tissues were determined. RESULTS: A total lavage volume of approximately 30 ml/kg x b.w. was used, and the liquid volume remaining in the lungs after operation ranged from 80 to 120 ml. All the dogs manifested significant declination in pulmonary function after the drowning. PaO2 in group B decreased significantly soon after the lavage but then increased gradually to a higher levels than group A. The dogs in group B had smaller lung water content and lower levels of lactate dehydrogenase and alkaline phosphatase in the bronchoalveolar fluid, showing improved oxygenation in comparison with group A. The inflammatory reaction and edema in lungs were also markedly alleviated in group B. CONCLUSION: Treatment with massive lung lavage in early stage after sea water aspiration can effectively protect the lung tissues from damage and alleviate the inflammatory reaction and pulmonary edema, helping to improve the lung function. PMID- 14559696 TI - [Effects of the antibacterial peptide cecropins from Chinese oak silkworm, Antheraea pernyi on 1, 2-dimethylhydrazine-induced colon carcinogenesis in rats]. AB - OBJECTIVE: To gain insight into the putative anticancer effect of the antibacterial peptides, cecropins, from Chinese oak silkworm, Antheraea pernyi, on the cancer cells and 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis in rats. METHODS: Growth inhibitory effect of the cecropins on normal human gastric epithelial cell line (GES-1) and human colon adenocarcinoma cell line (LS 174T) was observed using a microculture tetrazolium (MTT) colorimetric methods. Male Wistar rats were divided into 4 groups. Group1 was given on a weekly basis cecropins from Antheraea pernyi (3,000 Ua/ml) by gavage at 2 doses of 10 ml/kg body weight and exposed to subcutaneous injection of DMH at the dose of 20 mg/kg body weight. Group 2 was received weekly DMH only. Group 3 was given the cecropins by gavage at the same dose as in group 1. Group 4 was weekly exposed to subcutaneous injection of EDTA (1 mmol/L). All treatments were completed in a course of 18 weeks and the experiment was finished at week 33. RESULTS: MTT assay showed selective cytotoxic activity of the cecropins against the human colon adenocarcinoma cells line. The viability of the cancer cells was about 54% and 100% for the normal cells. There was a significantly lower incidence of large intestinal tumors in rats gavaged with cecropins (65%, P<0.01), but the tumor burden (tumors/tumor-bearing animal) and tumor mass index were comparable between the groups (P>0.05). CONCLUSION: The cecropins possess effective anti-tumor activity with no cytotoxicity against normal eukaryotic cells, and impede the neoplastic process in murine large intestines. PMID- 14559697 TI - [Effects of Huobahuagen tablet on the expression of interleukin-3 receptor mRNA in asthmatic guinea pig bronchoalveolar lavage fluid and eosinophil infiltration]. AB - OBJECTIVE: To investigate the effects of Huobahuagen tablet on the expression of interleukin-3 (IL-3) receptor mRNA in bronchoalveolar lavage fluid (BALF) and infiltration of eosinophil (Eos) in the airway of asthmatic guinea pigs. METHODS: Thirty-two healthy guinea pigs were randomized into 4 equal groups, the control group, asthmatic group, dexamethasone therapy group and Huobahuagen tablet therapy group. Asthma was induced in the latter 3 groups which were challenged with the asthma-inducing agents and at the same time received treatments as indicated. BALF were collected subsequently from the guinea pigs for examining the total cell number and cell classification, and histopathologic examination of the lung tissue was performed. Semi-quantitative analysis with reverse transcriptional-polymerase chain reaction (RT-PCR) of IL-3 receptor mRNA in the BALF was performed. RESULTS: Compared with the control and the asthmatic group, the number of Eos in the BALF of Huobahuagen tablet therapy group was significantly lower (P<0.05 and P<0.01, respectively). In spite of the presence of hyperemia and edema in the lung tissues of the Huobahuagen tablet therapy group, Eos infiltration was less severe than that in the asthmatic group. As found by RT-PCR, the quantity of IL-3 receptor mRNA in the BALF of the Huobahuagen tablet therapy group did not significantly differ from that in the dexamethasone therapy group, but was significantly higher than that in both the control and asthmatic group (P<0.01). CONCLUSION: Huobahuagen tablet can significantly lower the number of Eos in the airway of asthmatic guinea pigs, a finding that may potentially serve as the elementary theoretical basis for clinical therapy of asthma. PMID- 14559698 TI - [Effects of nitric oxide synthase inhibitor on dentate gyrus neurogenesis after diffuse brain injury in adult rats]. AB - OBJECTIVE: To investigate the effects of selective nitric oxide synthase (NOS) inhibitors on dentate gyrus (DG) neurogenesis after diffuse brain injury (DBI) in adult rats. METHODS: DBI models were established in adult male SD rats, followed by systemic bromodeoxyuridine (BrdU) labeling of the dividing cells and immunohistochemical assay of the proliferation rates of neural precursor cells in the DG for comparison between NOS inhibitor (7-nitroindazole and Aminoguanidine) groups and the corresponding control groups at various time points after DBI. RESULTS: Intraperitoneal administration of 7-nitroindazole significantly reduced the number of BrdU-labeled cells in the DG of adult rats 2, 4 and 6 d after DBI (P<0.05). Aminoguanidine also significantly inhibited the proliferation of neural precursor cells in the DG induced by DBI at various time points (P<0.01). CONCLUSIONS: The activation of NOS after DBI may be an important regulatory factor for DG neurogenesis in adult rats, and NO generated by nNOS is probably involved mainly in the early stage of enhanced neurogenesis after DBI, while the NO from iNOS might participated primarily in the later stages. PMID- 14559699 TI - [Clinical feature and surgical treatment of recurrent craniopharyngiomas]. AB - OBJECTIVE: To explore the clinical feature and surgical treatment of recurrent craniopharyngiomas. METHODS: The imaging features, intraoperative course, and postoperative outcomes of 83 patients with craniopharyngiomas were retrospectively analyzed. RESULTS: Of the 81 patients that survived, 19 experienced recurrence of the craniopharyngioma 6 to 62 months after the completion of the initial treatment, with recurrence rates of patients with total, subtotal and partial removal being 5.3%, 52.9%, and 100% respectively. Five of the 24 patients aged 6 years or older, who had an incomplete surgical removal, received radiation therapy, and only 1 of them experienced tumor recurrence (20%) whereas as many as 15 of the 19 patients (78.9%) did who did not received it. CONCLUSION: Surgery resection should be the primary therapeutic option for patients with recurrent craniopharyngioma and radiation therapy should also be considered but only as adjuvant therapy. PMID- 14559700 TI - [Changes of oxygen cost of breathing in children and adult during anesthesia and surgery]. AB - OBJECTIVE: To investigate the changes of oxygen cost of breathing (OCB) in children and adult during general anesthesia and surgery. METHODS: This study included 12 elective surgical children and 12 adult patients receiving cerebral surgery. Mechanical ventilation was given during general anesthesia and surgery, and the indices of hemodynamics and respiratory function were measured at 5 min before induction, 1 min before the start of surgery, 30, 60, 120 and 180 min after the start of surgery and at the end of surgery, respectively. RESULTS: Mean arterial pressure (MAP) in the 2 groups did not undergo any conspicuous changes during general anesthesia and surgery (P>0.05), and the heart rate in child group was markedly faster than that in adult group during the whole course (P<0.01). Oxygen consumption (VO2) in 2 groups at 1 min before the start of surgery and 30 min after the start of surgery was significantly higher than that at 5 min before induction (P<0.01), and at these two time points after induction, child group had much higher VO2 (P<0.05) but evidently lower OCB (P<0.01) than in adult group, the latter index remained low till 60 and 120 min after the start of surgery (P<0.05). CONCLUSION: During anesthesia and surgery, VO2 in child group can be much greater than that in Adult group, while OCB markedly lower. PMID- 14559701 TI - [Detection and analysis of SARS coronavirus-specific antibodies in sera from non SARS children]. AB - OBJECTIVE: To examine the presence of severe acute respiratory syndrome (SARS) coronavirus-specific antibodies in the sera from non-SARS children. METHODS: Indirect immunofluorescent assay and double-antigen sandwich enzyme-linked immunosorbent assay (ELISA) were used to detect the virus-specific antibodies in sera of 1,060 non-SARS children in Guangzhou. RESULTS: All the serum samples from the 1,060 non-SARS children were negative for both IgG and IgM antibodies against SARS coronavirus as determined by indirect immunofluorescent assay, with only two serum samples showing weak positivity for SARS coronavirus-specific antibodies identified by double-antigen sandwich ELISA. CONCLUSION: No SARS coronavirus specific antibody are present in the sera of non-SARS children. PMID- 14559702 TI - [Clinical observation of the efficacy of navelbine combined with ifosfamide and cisplatin in the treatment of advanced breast cancer]. AB - OBJECTIVE: To observe the efficacy of navelbine combined with ifosfamide and cisplatin in the treatment of advanced breast cancer. METHODS: Forty cases of pathologically confirmed stages III-IV breast cancer were treated with combined chemotherapy including intravenously infused navelbine (25 mg/m2, days 1 and 8), ifosfamide (1.3 g/m2, days 1 to 5) and cisplatin (20 mg/m2, days 1-5), administered in cycles of 21 days. The therapeutic effect was assessed after at least two cycles were completed. RESULTS: Complete remission was achieved in 6 cases and partial remission in 16, with an overall responding rate of 55%. The main toxicity was bone marrow suppression manifested by grades III-IV leukopenia occurring at a rate of 40%. CONCLUSION: Navelbine in combination with ifosfamide and cisplatin is effective and safe in the treatment of advanced breast cancer. PMID- 14559703 TI - [Association between the survival time and high-expression of EGFR and HER-2 in breast cancer]. AB - OBJECTIVE: To investigate the relationship between the survival time and the high expression of epidermal growth factor receptor (EGFR) and human epidermal receptor 2 (HER2) in breast cancer patients, and assess the feasibility of using the two markers either alone or in combination for predicting the prognosis of the patients. METHODS: Breast cancer samples were obtained from 185 patients and measured for the expressions of EGFR and HER-2 by way of immunohistochemistry, and 120 patients (64.9%) were followed up and their survival time recorded. Sixty five patients (35.1%) failed to be followed for various reasons. RESULTS: Of the 120 patients followed up, death occurred in 28 (15%). Positive HER2 expression was detected in 57.8% and EGFR expression in 40.5% of the all the samples examined. The over-expression of either HER2 or EGFR was in inverse correlation with the survival time (P<0.05 and P<0.01, respectively), and the over-expression of both related to the survival time in similar manner (P<0.05 and P<0.01). CONCLUSION: The high expression of HER2 or/and EGFR suggests a short survival time and an unfavorable prognosis. PMID- 14559704 TI - [Radiofrequency thermal ablation combined with arterial embolization for primary liver cancer]. AB - OBJECTIVE: To explore the short-term effect of radiofrequency ablation (RFA) combined with arterial embolization in treating patients with primary liver cancer. METHODS: Thirty patients with primary liver cancer received the combined treatment and the pre- and post-operative alpha-fetoprotein (AFP) levels, imaging features, and liver function were investigated along with observation of the incidence of complications to evaluate the therapeutic effects. RESULTS: The post operative AFP positivity and the tumor density were significantly reduced in these patients, and their one-year survival rate reached 96.7% with only minor complications observed. CONCLUSION: RFA combined with arterial embolization is effective for primary liver cancer. PMID- 14559705 TI - [Detection of platelet activation by flow cytometry in patients with pre eclampsia]. AB - OBJECTIVE: To explore the clinical significance of expression alterations of static and activated platelet glycoprotein CD41a, CD62p, and CD63 in patients with pre-eclampsia. METHODS: We used non-washing immunolabeling technique for fluorescence labeling of the static and activated platelet with CD41a, CD62p, and CD63 monoclonal antibodies of the blood samples from 16 patients with pre eclampsia, 15 women with normal pregnancies and 20 healthy subjects. The expression of the monoclonal antibody was detected by flow cytometry. RESULTS: The average static and activated platelet glycoprotein CD41a, CD62p, and CD63 expression levels in patients with pre-eclampsia were apparently higher than those of the normal pregnant women (P<0.01), while the comparison between the latter and the control group did not reveal any significant differences (P>0.05). CONCLUSION: High levels of platelet glycoprotein CD62p and CD63 expressions are present in patients with preeclampsia, suggesting the presence of platelet activation in these patients. PMID- 14559706 TI - [Analysis of alkaline phosphatase staining results in 238 cases of malignant lymphoma]. AB - OBJECTIVE: To examine the significance of alkaline phosphatase (ALP) staining in clinical characterization of malignant lymphoma cases. METHODS: Fresh peripheral blood smear obtained from 238 cases of malignant lymphoma, including 23 cases with bone marrow involvement (BMI) 189 cases without BMI(NBMI), and 26 cases of lymphoma cell leukemia (LCL), were examined by ALP staining, and the results were compared with those of 40 normal control subjects. RESULTS: With the gradual progression of lymphoma, the patients tended to have higher ALP scores, and the percentages of the subjects with a score higher than 250 in the control, NBMI, BMI and LCL were 0, 4.8%, 39.1%, 53.9% respectively, showing significant differences between the four groups. CONCLUSIONS: Increased ALP activity can be indirectly indicative of lymphoma progression, and ALP staining results may provide insight into the clinical staging and prognosis of malignant lymphoma. PMID- 14559707 TI - [Resistance of Neisseria gonorrhoeae to 4 antibiotics: an analysis of the situation in Guangzhou in 2002]. AB - OBJECTIVE: To survey the antibiotic resistance and the prevalence of penicillinase-producing and highly tetracycline-resistant Neisseria gonorrhoeae (PPNG and TRNG) in Guangzhou in 2002. METHODS: Minimum inhibitory concentrations (MIC) of the 4 antimicrobial agents, tetracycline, spectinomycin, ciprofloxacin and ceftriaxone, to 100 strains of Neisseria gonorrhoeae were determined by agar dilution method, and beta-lactamase was detected by iodine quantification method. RESULTS: There were 15 (15%) strains of PPNG and 28 (28%) strains of TRNG among the 100 strains of Neisseriae gonorrhoeae examined, with a resistant rate to ciprofloxacin as high as 98% and 32 (32%) strains identified to be highly resistant to the antibiotics (MIC >or=16 mg/L). No resistant strains to ceftriaxone and specitinomycin were identified, but ceftriaxone susceptibility of the strains was declining. CONCLUSIONS: The current antibiotic resistance of gonococci strains are still serious in Guangzhou, especially in the case of ciprofloxacin resistance. The strains are sensitive to spectinomycin and ceftriaxone, and continuous monitoring of the bacteria for their antibiotic resistance is necessary. PMID- 14559709 TI - [Study of the extraction technique of volatile oil from Chinese herbs for manufacturing Xiaoerjianpixiaoshi oral solution]. AB - OBJECTIVE: To determine the optimal extracting technique for the volatile oil from the Chinese herbs for manufacturing Xiaoerjianpixiaoshi oral solution. METHODS: Two steam distillation processes were carried out to extract the volatile oil from three Chinese herbs. UV-spectrophotometry method was used to determine the oil content in the distillate, and the mathematical model was established according to volatile oil distillation process by exponential curve regression analysis. RESULT: Direct extraction of the oil by means of steam distillation process was more workable, and the decrement of oil content conformed well with the exponential model. Collection of 2 volumes of the distillate resulted in the yield of volatile oil of 61.64%. CONCLUSION: The optimal extraction of the volatile oil can be resulted from immersion of the drugs in 8 volumes of water for 0.5 h and collection of 2 volumes of the distillate. PMID- 14559708 TI - [Large-scale real-time titration of green-fluorescence-protein-marked recombinant retrovirus: comparison with standard titration method]. AB - OBJECTIVE: To compare large-scale real-time titration method (LaSRT) with standard titration method by flow cytometry (FACS) for determining the titers of green-fluorescence-protein (GFP)-marked recombinant retrovirus. METHODS: (1) Standard titration method: NIH3T3 cells were inoculated at 2x10(5) /well in 6 well plate, and after cell culture for 12 h, 0.5 ml, 50 microl and 5 microl GFP marked recombinant retrovirus (n=3) were respectively used to infect the cells, with the final concentration of polybrene being 8 microg/ml. Forty-eight hours later, the cells were treated with trypsin and assayed for the positive rate of GFP by means of FACS. When the positive rate was lower than 10%, the titer was calculated according to the equation: virus titer (TU/ml) =2x10(5)xGFP positive rate/volume of virus stock solution used. (2) LaSRT method: The cells were inoculated at 5,000 cells/well in a 96-well plate, and after cell culture for 12 h, 90 microl/well complete culture medium was used with 8 microg/ml polybrene and 10% newborn bovine serum, 10 microl virus was added into the first well, and ten fold dilution of the previous virus-containing solution was performed before the virus was added into the next well (8 wells in each group, altogether 3 groups). Forty-eight hours later, inverted fluorescence microscope was used to observe the fluorescence-positive cells in each well. The virus titer was calculated according to the equation: virus titer (TU/ml) =mx10(n+1), where n is the serial number of the reference well, and m the number of positive cells. (3) LaSRT was used to study the influence of freezing/thawing on the titers of recombinant retroviruses. RESULTS: The virus titer obtained with standard method by FACS was (1.54+/-0.38)x10(6) TU/ml, and that of LaSRT was (1.33+/-0.57)x10(6) TU/ml (P>0.05). After one cycle of freezing/thawing, the virus titer dropped to (18.1+/ 9.9)% (n=7). CONCLUSION: LaSRT is more rapid and convenient as well as easier to determine the virus titer compared with standard method, and no significant difference is found between the two titration methods. PMID- 14559710 TI - [On "the heart also governs Qi"]. AB - As the organ of supreme superiority over any other organ of the body, the heart controls the function of the whole body. The lung governs the motions of Qi(vital energy), as have long been established in the basic theories in traditional Chinese medicine, but in this paper, the authors attempt to define the close relations between the heart and Qi by proposing that the heart also governs Qi, through either immediate and indirect actions. The heart plays an important role in the motion, generation and function regulation of Qi, with also impact on the changes of Qi ji, it is therefore of great importance to obtain a thorough comprehension of the associations between the two facets of Qi government by the lung and the heart, for the purpose of accurate clinical identification of the diseases and consequently appropriate treatment. PMID- 14559711 TI - [Intravascular administration of pingyangmycin lipiodol emulsion for Klippel Trenaunay syndrome: clinical analysis of 7 cases]. AB - OBJECTIVE: To define the arteriographic features of Klippel-Trenaunay sydrome (KTS) and explore the feasibility of the treatment with intravascular administration of pingyangmycin lipiodol emulsion (PLE). METHODS: Seven patients with KTS underwent arteriography followed by therapy with PLE administration (containing pingyangmycin 8-16 mg and lipiodol 4-6 ml) into the trunk of the arteries of all impaired extremities. The therapeutic effect and postoperative complications were observed. RESULTS: Arteriography revealed abundant small branches of the runoff arteries, showing sinusoid changes of varied sizes in 5 cases by staining of the soft tissue. During the 3 to 30-month follow-up, swelling of the extremities was obviously improved in 5 cases after PLE therapy, which failed to produce obvious effect in 1 case and recurrence was detected in another. The major complications included paresthesia of the limbs, serum transaminase disorder and local skin necrosis. CONCLUSION: Arteriography can detect the lesions of the arteries and soft tissues in the affected extremity of patients with KTS, intravascular administration of PLE was effective to alleviate the hypertrophy of the limbs. PMID- 14559712 TI - [Change of protein catabolism and immunocompetence in severely burned patients after operations and the therapeutic effect of recombinant human growth hormone]. AB - OBJECTIVE: To investigate the changes in protein catabolism and immunocompetence in severely burned patients after operations and observe the therapeutic effect of recombinant human growth hormone (rhGH) in these patients. METHODS: The changes in plasma protein concentration and immune function and the clinical results of 38 severely burned patients after three operations were retrospectively analyzed. These patients were assigned to GroupI(control, n=20) and GroupII (rhGH treatment, n=18), and matched for age, total body surface area (TBSA), surgical approaches and times, perioperative bleeding volume and nutrition support etc. The patients in groupII received intramuscular injection with 0.2-0.4 U/d x kg x b.w. rhGH after the first operation, till the healing of the wounds. RESULTS: The concentration of retina-binding protein, transferrin, prealbumin, albumin, and IgG, the number of CD4 and CD8 cells, as well as the ratio of CD4/CD8 cells prior to the operation were below the normal levels in the two groups of patients. All these indexes in GroupII recovered to a great extent after three operations and were significantly higher than those in GroupI(P<0.05). CONCLUSION: Nutritious state and immunocompetence of patients with severe burns are severely compromised after operations, and nutrition support and rhGH treatment may produce rapid improvement of the patients' conditions. PMID- 14559713 TI - [Lung transplantation for treatment of pulmonary cystic fibrosis: report of one case]. AB - A right single lung transplantation was performed in a 17-year-old female patient with end-stage cystoid pulmonary fibrosis and without cardiopulmonary bypass in June 2003. The donor lung was perfused with cold UW solution with a cold ischemic time of 280 min. The patient weaned from ventilator on the next day of operation (18 h later) and was able to walk at the fourth day postoperatively. Immunosuppression included methylprednisolone used before FK 506, mycophenolate mofetil and prednisone dosed after operation. The patient remains well a month after operation with significant improvement of the lung function and enjoys normal life. PMID- 14559714 TI - Hexosamines and TGF-beta1 use similar signaling pathways to mediate matrix protein synthesis in mesangial cells. AB - Hyperglycemia-induced alterations in mesangial (MES) cell function and extracellular matrix (ECM) protein accumulation are seen in diabetic glomerulopathy. Transforming growth factor-beta1 (TGF-beta1) mediates high glucose-induced matrix production in the kidney. Recent studies demonstrated that some of the effects of high glucose on cellular metabolism are mediated by the hexosamine biosynthesis pathway (HBP) in which fructose-6-phosphate is converted to glucosamine (GlcN) 6-phosphate. We previously showed that the high-glucose mediated fibronectin and laminin synthesis in MES cells is mediated by the HBP and that GlcN is more potent than glucose in inducing TGF-beta1 promoter luciferase activity. In this study, we investigated the hypothesis that the effects of glucose on MES matrix production occur via hexosamine regulation of TGF-beta1. Culturing simian virus (SV)-40-transformed rat kidney MES cells in 25 mM glucose (HG) for 48 h increases cellular fibronectin and laminin levels about twofold on Western blots compared with low glucose (5 mM). GlcN (1.5 mM) or TGF beta1 (2.5-5 ng/ml) for 24-48 h also increases ECM synthesis. However, the effects of HG or GlcN with TGF-beta1 are not additive. The presence of anti-TGF beta1 antibodies (20 microg/ml) blocks both TGF-beta1- and GlcN-induced fibronectin synthesis. TGF-beta1 increased ECM levels via PKA (laminin and fibronectin) and PKC (fibronectin) pathways. Similarly, TGF-beta1 and hexosamines led to nonadditive increases in phosphorylation of the cAMP responsive element binding transcription factor. These results suggest that the effects of excess glucose on MES ECM synthesis occur via HBP-mediated regulation of TGF-beta1. PMID- 14559715 TI - Dietary salt intake modulates progression of antithymocyte serum nephritis through alteration of glomerular angiotensin II receptor expression. AB - Dietary salt intake modulates the renin-angiotensin system (RAS); however, little is known about the effect of salt intake on the progression of glomerulonephritis. We investigated the glomerular expression of TGF-beta1 type I (TbetaRI) and II (TbetaRII) TGF-beta receptors and RAS components in rats with antithymocyte serum (ATS) nephritis on normal (NSI)-, low (LSI)-, and high-salt intake (HSI) and on HSI rats receiving candesartan cilexetil (CC) and LSI rats receiving PD-123319. Glomerular lesions were less severe in rats on LSI and aggravated in those on HSI compared with those on NSI. Intrarenal renin and glomerular ANG II levels were significantly higher in LSI and lower in HSI rats. In ATS nephritis, HSI increased glomerular TbetaRI, TbetaRII, and ANG II type 1 receptor (AT1R), and decreased glomerular ANG II type 2 receptor (AT2R), whereas LSI decreased glomerular TGF-beta1 and TbetaRI and increased glomerular AT2R. CC ameliorated glomerular lesions, reduced glomerular TGF-beta1 and TbetaRII, and increased glomerular AT2R. PD-123319 aggravated glomerular lesions and increased glomerular TGF-beta1 and TbetaRII. Our results suggest that dietary salt intake influences progression of ATS nephritis by modulating glomerular TGF-beta1 and TbetaR expression resulting, at least in part, from altered glomerular AT1R and AT2R expression. PMID- 14559716 TI - Decreased amiloride-sensitive Na+ absorption in collecting duct principal cells isolated from BPK ARPKD mice. AB - The main feature of polycystic kidney diseases (PKD) is formation and progressive enlargement of renal cysts. Alterations in epithelial cell proliferation, extracellular matrix, and ion transport are thought to contribute to cyst enlargement and loss of renal function. Abnormal Cl- secretion is implicated in cyst enlargement in autosomal dominant PKD (ADPKD), but little is known about transport abnormalities in autosomal recessive PKD (ARPKD). We developed a method to isolate collecting duct (CD) principal cells (site of the lesion in ARPKD) from normal and ARPKD mice. A transgenic mouse (Hoxb7/GFP) in which enhanced green fluorescent protein (GFP) is expressed in CDs was bred with an ARPKD mouse (BPK), and GFP-positive cells from normal and cystic mice were selected by fluorescence-activated cell sorting. GFP-positive CD cells (>95 +/- 3%) obtained from either normal or cystic mice formed high-resistance, polarized epithelial monolayers. Expression patterns for marker proteins and the presence of a central cilium confirmed that the monolayers are composed of principal cells. Under basal conditions, the Cl- secretory responses elicited by elevation of cAMP or calcium were not significantly different between normal and cystic monolayers. In contrast, the amiloride-sensitive short-circuit current was significantly reduced in monolayers of cells isolated from cystic mice (12.9 +/- 1.6 microA/cm2; n = 10) compared with monolayers of cells isolated from normal mice (27.3 +/- 3.4 microA/cm2; n = 12). The results of these studies suggest that epithelial sodium channel-mediated sodium absorption is decreased in principal cells of ARPKD CD cysts and that the reduction in sodium absorption may contribute to the accumulation of luminal fluid. PMID- 14559717 TI - Epidermal growth factor decreases PEPT2 transport capacity and expression in the rat kidney proximal tubule cell line SKPT0193 cl.2. AB - The renal peptide transporter PEPT2 plays an important role in absorption of di- and tripetides in the proximal tubule; however, knowledge of regulation of PEPT2 by growth factors and hormones is limited. In the present study, we examined the effects of epidermal growth factor (EGF) on PEPT2 transport capacity and expression in the rat proximal tubule cell line SKPT0193 cl.2 (SKPT), which expresses rat PEPT2 (rPEPT2) in the apical membrane. Treatment of SKPT cells with EGF during cell culture growth caused a dose-dependent decrease in rPEPT2 transport capacity and expression, as determined by studies of apical uptake of [14C]glycylsarcosine, rPepT2 mRNA levels, and immunostaining of SKPT cells with a rPEPT2-specific antibody. On the contrary, apical uptake of glucose and lysine was increased in EGF-treated cells, indicating that EGF was not acting generally to decrease apical nutrient uptake mechanisms in the proximal tubule cells. Our findings indicate that EGF decreases rPEPT2 expression by lowering transcription of the rat PepT2 gene or by decreasing rat PepT2 mRNA stability. Previous investigators routinely used SKPT cell culture media with a high (10 ng/ml) EGF concentration. Our study suggests that this might be disadvantageous when studying PEPT2-mediated transport phenomena. These findings demonstrate for the first time EGF-mediated regulation of PEPT2 expression in a kidney cell line. The relevance for kidney regulation of peptide transport activity in physiological and/or pathophysiological situations, where EGF and EGF receptor levels change drastically, remains to be established. PMID- 14559718 TI - Extracellular matrix regulates glomerular epithelial cell survival and proliferation. AB - Glomerular epithelial cell (GEC) injury and apoptosis may contribute to sclerosis in glomerulonephritis. The present study addresses signals that regulate survival of GEC in culture and in the acute puromycin aminonucleoside nephrosis (PAN) model of GEC injury in vivo. Compared with GEC on plastic substratum, adhesion to collagen increased activation of focal adhesion kinase (FAK), c-Src, and ERK and facilitated survival (prevented apoptosis). GEC on plastic exhibited increased caspase-8 and -9 activities, increased expression of the proapoptotic protein, Bax, and decreased the antiapoptotic protein, Bcl-XL, compared with collagen. Stable expression of constitutively active mutants of FAK (CD2-FAK) or MEK (R4F MEK) activated the ERK pathway and supplanted the requirement of collagen for survival. In contrast, expression of a Ras mutant that activates phosphatidylinositol 3-kinase but blocks ERK activation or pharmacological inhibition of the ERK pathway decreased survival on collagen. Glomeruli isolated from rats with PAN revealed increased beta1-integrin expression, along with increased activation of FAK, c-Src, and ERK, compared with controls. EGF receptor activation was undetectable in PAN. Therefore, adhesion to collagen, resulting in activation of FAK and the Ras-ERK pathway, supports GEC survival. Analogous signals for GEC survival are activated in PAN. PMID- 14559719 TI - Expression profiling of the gamma-subunit isoforms of AMP-activated protein kinase suggests a major role for gamma3 in white skeletal muscle. AB - Expression patterns of the three isoforms of the regulatory gamma-subunit of AMP activated protein kinase (AMPK) were determined in various tissues from adult humans, mice, and rats, as well as in human primary muscle cells. Real-time PCR based quantification of mRNA showed similar expression patterns in the three species and a good correlation with protein expression in mice and rats. The gamma3-isoform appeared highly specific to skeletal muscle, whereas gamma1 and gamma2 showed broad tissue distributions. Moreover, the proportion of white, type IIb fibers in the mouse and rat muscle samples, as indicated by real-time PCR quantification of Atp1b2 mRNA, showed a strong positive correlation with the expression of gamma3. In samples of white skeletal muscle, gamma3 clearly appeared to be the most abundant gamma-isoform. Differentiation of human primary muscle cells from myoblasts into multinucleated myotubes was accompanied by upregulation of gamma3 mRNA expression, whereas levels of gamma1 and gamma2 remained largely unchanged. However, even in these cultured myotubes, gamma2 was the most highly expressed isoform, indicating a considerable difference compared with adult skeletal muscle. Immunoblot analysis of mouse gastrocnemius and quadriceps muscle extracts precipitated with a gamma3-specific antibody showed that gamma3 was exclusively associated with the alpha2- and beta2-subunit isoforms. The observation that the AMPKgamma3 isoform is expressed primarily in white skeletal muscle, in which it is the predominant gamma-isoform, strongly suggests that gamma3 has a key role in this tissue. PMID- 14559720 TI - Antenatal glucocorticoids reset the level of baseline and hypoxemia-induced pituitary-adrenal activity in the sheep fetus during late gestation. AB - This study examined the effects of dexamethasone treatment on basal hypothalamo pituitary-adrenal (HPA) axis function and HPA responses to subsequent acute hypoxemia in the ovine fetus during late gestation. Between 117 and 120 days (term: approximately 145 days), 12 fetal sheep and their mothers were catheterized under halothane anesthesia. From 124 days, 6 fetuses were continuously infused intravenously with dexamethasone (1.80 +/- 0.15 microg.kg( 1).h(-1) in 0.9% saline at 0.5 ml/h) for 48 h, while the remaining 6 fetuses received saline at the same rate. Two days after infusion, when dexamethasone had cleared from the fetal circulation, acute hypoxemia was induced in both groups for 1 h by reducing the maternal fraction of inspired O2. Fetal dexamethasone treatment transiently lowered fetal basal plasma cortisol, but not ACTH, concentrations. However, 2 days after treatment, fetal basal plasma cortisol concentration was elevated without changes in basal ACTH concentration. Despite elevated basal plasma cortisol concentration, the ACTH response to acute hypoxemia was enhanced, and the increment in plasma cortisol levels was maintained, in dexamethasone-treated fetuses. Correlation of fetal plasma ACTH and cortisol concentrations indicated enhanced cortisol output without a change in adrenocortical sensitivity. The enhancements in basal cortisol concentration and the HPA axis responses to acute hypoxemia after dexamethasone treatment were associated with reductions in pituitary and adrenal glucocorticoid receptor mRNA contents, which persisted at 3-4 days after the end of treatment. These data show that prenatal glucocorticoids alter the basal set point of the HPA axis and enhance HPA axis responses to acute stress in the ovine fetus during late gestation. PMID- 14559721 TI - High-fat diet elevates resting intramuscular triglyceride concentration and whole body lipolysis during exercise. AB - This study determined the role of intramuscular triglyceride (IMTG) and adipose lipolysis in the elevated fat oxidation during exercise caused by a high-fat diet. In four separate trials, six endurance-trained cyclists exercised at 50% peak O2 consumption for 1 h after a two-day control diet (22% fat, CON) or an isocaloric high-fat diet (60% fat, HF) with or without the ingestion of acipimox, an adipose lipolysis inhibitor, before exercise. During exercise, HF elevated fat oxidation by 72% and whole body lipolysis [i.e., the appearance rate of glycerol in plasma (Ra glycerol)] by 79% compared with CON (P < 0.05), and this was associated with a 36% increase (P < 0.05) in preexercise IMTG concentration. Although acipimox lowered plasma free fatty acid (FFA) availability, HF still increased fat oxidation and Ra glycerol to the same magnitude above control as the increase caused by HF without acipimox (i.e., both increased fat oxidation 13 14 micromol.kg(-1).min(-1)). In conclusion, the marked increase in fat oxidation after a HF diet is associated with elevated IMTG concentration and whole body lipolysis and does not require increased adipose tissue lipolysis and plasma FFA concentration during exercise. This suggests that altered substrate storage in skeletal muscle is responsible for increased fat oxidation during exercise after 2 days of an HF diet. PMID- 14559723 TI - Induction of control genes in intestinal gluconeogenesis is sequential during fasting and maximal in diabetes. AB - We studied in rats the expression of genes involved in gluconeogenesis from glutamine and glycerol in the small intestine (SI) during fasting and diabetes. From Northern blot and enzymatic studies, we report that only phosphoenolpyruvate carboxykinase (PEPCK) activity is induced at 24 h of fasting, whereas glucose-6 phosphatase (G-6-Pase) activity is induced only from 48 h. Both genes then plateau, whereas glutaminase and glycerokinase strikingly rebound between 48 and 72 h. The two latter genes are fully expressed in streptozotocin-diabetic rats. From arteriovenous balance and isotopic techniques, we show that the SI does not release glucose at 24 h of fasting and that SI gluconeogenesis contributes to 35% of total glucose production in 72-h-fasted rats. The new findings are that 1) the SI can quantitatively account for up to one-third of glucose production in prolonged fasting; 2) the induction of PEPCK is not sufficient by itself to trigger SI gluconeogenesis; 3) G-6-Pase likely plays a crucial role in this process; and 4) glutaminase and glycerokinase may play a key potentiating role in the latest times of fasting and in diabetes. PMID- 14559722 TI - Prolactin, progesterone, and dexamethasone coordinately and adversely regulate glucokinase and cAMP/PDE cascades in MIN6 beta-cells. AB - Islet cells undergo major changes in structure and function to meet the demand for increased insulin secretion during pregnancy, but the nature of the hormonal interactions and signaling events is incompletely understood. Here, we used the glucose-responsive MIN6 beta-cell line treated with prolactin (PRL), progesterone (PRG), and dexamethasone (DEX, a synthetic glucocorticoid), all elevated during late pregnancy, to study their effects on mechanisms of insulin secretion. DEX alone or combined with PRL and PRG inhibited insulin secretion in response to 16 mM glucose-stimulating concentrations. However, in the basal state (3 mM glucose), the insulin levels in response to DEX treatment were unchanged, and the three hormones together maintained higher insulin release. There were no changes of protein levels of GLUT2 or glucokinase (GK), but PRL or PRG treatment increased GK activity, whereas DEX had an inhibitory effect on GK activity. alpha Ketoisocaproate (alpha-KIC)-stimulated insulin secretion was also reduced by DEX alone or combined with PRL and PRG, suggesting that DEX may inhibit distal steps in the insulin-exocytotic process. PRL treatment increased the concentration of intracellular cAMP in response to 16 mM glucose, suggesting a role for cAMP in potentiation of insulin secretion, whereas DEX alone or combined with PRL and PRG reduced cAMP levels by increasing phosphodiesterase (PDE) activity. These data provide evidence that PRL and to a lesser extent PRG, which increase in early pregnancy, enhance basal and glucose-stimulated insulin secretion in part by increasing GK activity and amplifying cAMP levels. Glucocorticoid, which increases throughout gestation, counteracts only glucose-stimulated insulin secretion under high glucose concentrations by dominantly inhibiting GK activity and increasing PDE activity to reduce cAMP levels. These adaptations in the beta cell may play an important role in maintaining the basal hyperinsulinemia of pregnancy while limiting the capacity of PRL and PRG to promote glucose stimulated insulin secretion during late gestation. PMID- 14559724 TI - Effect of high-intensity intermittent training on lactate and H+ release from human skeletal muscle. AB - The study investigated the effect of training on lactate and H+ release from human skeletal muscle during one-legged knee-extensor exercise. Six subjects were tested after 7-8 wk of training (fifteen 1-min bouts at approximately 150% of thigh maximal O2 uptake per day). Blood samples, blood flow, and muscle biopsies were obtained during and after a 30-W exercise bout and an incremental test to exhaustion of both trained (T) and untrained (UT) legs. Blood flow was 16% higher in the T than in the UT leg. In the 30-W test, venous lactate and lactate release were lower in the T compared with the UT leg. In the incremental test, time to fatigue was 10.6 +/- 0.7 and 8.2 +/- 0.7 min, respectively, in the T and UT legs (P < 0.05). At exhaustion, venous blood lactate was 10.7 +/- 0.4 and 8.0 +/- 0.9 mmol/l in T and UT legs (P < 0.05), respectively, and lactate release was 19.4 +/ 3.6 and 10.6 +/- 2.0 mmol/min (P < 0.05). H+ release at exhaustion was higher in the T than in the UT leg. Muscle lactate content was 59.0 +/- 15.1 and 96.5 +/- 14.5 mmol/kg dry wt in the T and UT legs, and muscle pH was 6.82 +/- 0.05 and 6.69 +/- 0.04 in the T and UT legs (P = 0.06). The membrane contents of the monocarboxylate transporters MCT1 and MCT4 and the Na+/H+ exchanger were 115 +/- 5 (P < 0.05), 111 +/- 11, and 116 +/- 6% (P < 0.05), respectively, in the T compared with the UT leg. The reason for the training-induced increase in peak lactate and H+ release during exercise is a combination of an increased density of the lactate and H+ transporting systems, an improved blood flow and blood flow distribution, and an increased systemic lactate and H+ clearance. PMID- 14559725 TI - Metabolic regulation of growth hormone by free fatty acids, somatostatin, and ghrelin in HIV-lipodystrophy. AB - Human immunodeficiency virus (HIV)-lipodystrophy is a syndrome characterized by changes in fat distribution and insulin resistance. Prior studies suggest markedly reduced growth hormone (GH) levels in association with excess visceral adiposity among patients with HIV-lipodystrophy. We investigated mechanisms of altered GH secretion in a population of 13 male HIV-infected patients with evidence of fat redistribution, compared with 10 HIV-nonlipodystrophic patients and 11 male healthy controls similar in age and body mass index (BMI). Although similar in BMI, the lipodystrophic group was characterized by increased visceral adiposity, free fatty acids (FFA), and insulin and reduced extremity fat. We investigated ghrelin and the effects of acute lowering of FFA by acipimox on GH responses to growth hormone-releasing hormone (GHRH). We also investigated somatostatin tone, comparing GH response to combined GHRH and arginine vs. GHRH alone with a subtraction algorithm. Our data demonstrate an equivalent number of GH pulses (4.1 +/- 0.6, 4.7 +/- 0.8, and 4.5 +/- 0.3 pulses/12 h in the HIV lipodystrophic, HIV-nonlipodystrophic, and healthy control groups, respectively, P > 0.05) but markedly reduced GH secretion pulse area (1.14 +/- 0.27 vs. 4.67 +/ 1.24 ng.ml(-1).min, P < 0.05, HIV-lipodystrophic vs. HIV-nonlipodystrophic; 1.14 +/- 0.27 vs. 3.18 +/- 0.92 ng.ml(-1).min, P < 0.05 HIV-lipodystrophic vs. control), GH pulse area, and GH pulse width in the HIV-lipodystrophy patients compared with the control groups. Reduced ghrelin (418 +/- 46 vs. 514 +/- 37 pg/ml, P < 0.05, HIV-lipodystrophic vs. HIV-nonlipodystrophic; 418 +/- 46 vs. 546 +/- 45 pg/ml, P < 0.05, HIV-lipodystrophic vs. control), impaired GH response to GHRH by excess FFA, and increased somatostatin tone contribute to reduced GH secretion in patients with HIV-lipodystrophy. These data provide novel insight into the metabolic regulation of GH secretion in subjects with HIV-lipodystrophy. PMID- 14559727 TI - Uncertainty and significance. PMID- 14559726 TI - Tracer-derived total and folate-dependent homocysteine remethylation and synthesis rates in humans indicate that serine is the main one-carbon donor. AB - Hyperhomocysteinemia in humans is associated with genetic variants of several enzymes of folate and one-carbon metabolism and deficiencies of folate and vitamins B12 and B6. In each case, hyperhomocysteinemia might be caused by diminished folate-dependent homocysteine remethylation, but this has not been confirmed in vivo. Because published stable isotopic tracer approaches cannot distinguish folate-dependent from folate-independent remethylation, we developed a dual-tracer procedure in which a [U-13C5]-methionine tracer is used in conjunction with a [3-13C]serine tracer to simultaneously measure rates of total and folate-dependent homocysteine remethylation. In young female subjects, plasma [U-13C4]homocysteine enrichment, a surrogate measure of intracellular [U 13C5]methionine enrichment, reached approximately 90% of the plasma [U 13C5]methionine enrichment. Methionine-methyl and -carboxyl group fluxes were in the range of previous reports (approximately 25 and approximately 17 micromol.kg( 1).h(-1), respectively). However, the rate of overall homocysteine remethylation (approximately 8 micromol.kg(-1).h(-1)) was twice that of previous reports, which suggests a larger role for homocysteine remethylation in methionine metabolism than previously thought. By use of estimates of intracellular [3-13C]serine enrichment based on a conservative correction of plasma [3-13C]serine enrichment, serine was calculated to contribute approximately 100% of the methyl groups used for total body homocysteine remethylation under the conditions of this protocol. This contribution represented only a small fraction (approximately 2.8%) of total serine flux. Our dual-tracer procedure is well suited to measure the effects of nutrient deficiencies, genetic polymorphisms, and other metabolic perturbations on homocysteine synthesis and total and folate-dependent homocysteine remethylation. PMID- 14559728 TI - Infectious diseases--past, present, and future. PMID- 14559729 TI - Tests of significance considered as evidence. Journal of the American Statistical Association 1942;37:325-35. PMID- 14559730 TI - Note on Dr. Berkson's criticism of tests of significance. 1943. PMID- 14559731 TI - Commentary: Null points--has interpretation of significance tests improved? PMID- 14559732 TI - Commentary: Worthwhile polemic or transatlantic storm-in-a-teacup? PMID- 14559733 TI - Commentary: The P-value, devalued. PMID- 14559735 TI - as if... visualizing pain. PMID- 14559737 TI - Commentary: Modelling and understanding differences in human immunodeficiency and hepatitis C virus epidemiology in injection drug users. PMID- 14559736 TI - The impact of behavioural changes on the prevalence of human immunodeficiency virus and hepatitis C among injecting drug users. AB - BACKGROUND: Prevalence of human immunodeficiency virus (HIV) and hepatitis C virus (HCV) in the injecting drug user (IDU) community can differ considerably. In Australia in 1997, HIV prevalence among attendees at Needle Exchange Programs was 1% while HCV prevalence was 50%. The impact that different needle-sharing behaviour and drug injecting use may have on the future levels of these viruses is uncertain. METHOD: We develop a mathematical model of the number of people who inject drugs with each of these infections to determine their changes under different scenarios. The impact of transmission probabilities and needle sharing on the incidence and prevalence of HIV and HCV infections are assessed. RESULTS: Critical levels of needle sharing, below which total infections would fall to minimal levels, were estimated to be 17 IDU partners per year for HIV compared with 3 IDU partners per year for HCV. Current average levels of needle sharing in Australia are estimated to be six IDU partners per year. CONCLUSIONS: This analysis suggests that under current drug injecting behaviour, HIV prevalence in IDU in Australia should remain below 1% but that HCV prevalence will stay elevated. PMID- 14559739 TI - Commentary: Modelling the epidemiology of hepatitis C and its complications. PMID- 14559738 TI - Modelling hepatitis C virus incidence, prevalence and long-term sequelae in Australia, 2001. AB - BACKGROUND: To plan an appropriate public health response to the hepatitis C virus (HCV) epidemic requires that estimates of HCV incidence and prevalence, and projections of the long-term sequelae of infection, are as accurate as possible. In this paper, mathematical models are used to synthesize data on the epidemiology and natural history of HCV in Australia to estimate HCV incidence and prevalence in Australia to end 2001, and project future trends in the long term sequelae of HCV infection. METHODS: Mathematical models of the HCV epidemic in Australia were developed based on estimates of the pattern of injecting drug use. Estimates of HCV infections due to injecting drug use were then adjusted to allow for HCV infections resulting from other transmission routes. Projections of the long-term sequelae of HCV infection were obtained by combining modelled HCV incidence with estimates of the progression rates to these outcomes. RESULTS: It was estimated that there were 210 000 (lower and upper limits of 157 000 and 252 000) people in Australia living with HCV antibodies at the end of 2001, with HCV incidence in 2001 estimated to be 16 000 (11 000-19 000). It was estimated that 6500 (5000-8000) people were living with HCV-related cirrhosis in 2001, that 175 (130-210) people developed HCV-associated liver failure, and that there were 50 (40-60) incident cases of HCV-related hepatocellular carcinoma (HCC). It was estimated that in 2001 22 500 quality adjusted life years were lost to chronic HCV infection, the majority (77%) in people with early (stage 0/1) liver disease. DISCUSSION: Model-based estimates were broadly consistent with other sources of information on the HCV epidemic in Australia. These models suggest that the prevalence of HCV-related cirrhosis and the incidence of HCV-related liver failure and HCC will more than triple in Australia by 2020. PMID- 14559740 TI - Influence of maternal human immunodeficiency virus (HIV) co-infection on vertical transmission of hepatitis C virus (HCV): a meta-analysis. AB - BACKGROUND: Maternal co-infection with human immunodeficiency virus (HIV) has been implicated as a potentially important co-factor for enhanced vertical transmission of hepatitis C virus (HCV). In previous reports, however, methodological issues (notably small sample sizes) have limited accurate evaluation of the contribution of maternal co-infection with HIV on the risk of vertical transmission of HCV. METHODS: A systematic review and subsequent meta analysis of current published and unpublished reports was performed. Odds ratios (OR) and 95% CI for individual studies were calculated with maternal HIV serostatus as the exposure measure and HCV vertical transmission as the outcome measure. Overall summary estimates were then calculated using a random effects model that estimates a weighted average of OR from individual studies. RESULTS: In total, 2382 infants from 10 studies were included in an analysis of HCV infected mothers (defined by anti-HCV+ antibody assays) with and without concomitant HIV infection. The risk estimate (OR) of HCV vertical transmission was 2.82 (95% CI: 1.78-4.45; P = 0.00001) from anti-HCV+/HIV+ co-infected mothers compared with anti-HCV+/HIV- mothers. In a subanalysis of 1327 infants born to viraemic (HCV RNA+) mothers, the risk estimate of HCV vertical transmission was 1.97 (95% CI: 1.04-3.74; P = 0.04) from HCV viraemic/HIV+ co-infected mothers compared with HCV viraemic/HIV- mothers. CONCLUSIONS: Results from this meta analysis of observational studies suggest that the risk of HCV vertical transmission is higher in infants born to HIV co-infected mothers. PMID- 14559741 TI - High human immunodeficiency virus type 1 seroprevalence in men who have sex with men in Buenos Aires, Argentina: risk factors for infection. AB - OBJECTIVE: To determine human immunodeficiency virus (HIV) seroprevalence in a sample of men who have sex with men (MSM) in Buenos Aires City and to identify risk factors associated with HIV type 1 infection. METHODS: Participants were invited to receive HIV counselling and testing at "NEXO" (a gay non-governmental organization) by means of informative leaflets distributed in gay nightclubs, porno cinemas, gymnasiums, and in the streets. During the encounter, the study was explained by a trained social worker and individuals were invited to volunteer for the study. Diagnosis of HIV was performed using two screening tests and Western Blot assay was used as confirmatory. RESULTS: Human immunodeficiency virus was detected in 96 (13.8%; 95% CI: 11.4-16.7) of 694 MSM. Fourteen (14.6%) of the 96 HIV-positive MSM were already aware of their HIV serostatus. In univariate analysis, HIV-1 infection (odds ratio [OR] >1.5) was found to be associated with older age (30-39 years), being unemployed, a previous sexually transmitted disease (STD) history, and having an HIV-positive partner. Cocaine consumption and irregular use of condoms with occasional partners were also found to be risk factors. In multivariate logistic regression analysis, being unemployed (OR = 3.42; 95% CI: 1.17-9.99) and having an HIV-positive partner (OR = 2.67; 95% CI: 1.09-6.52) remained significant risk factors. DISCUSSION: The high HIV-1 prevalence observed suggests an urgent need for implementation of effective prevention campaigns. This represents the first cross-sectional epidemiological study of HIV among the high-risk group of MSM in Argentina. PMID- 14559742 TI - Commentary: The human immunodeficiency virus/AIDS epidemic among men who have sex with men in Latin America and the Caribbean: it is time to bridge the gap. PMID- 14559743 TI - Mobility and the spread of human immunodeficiency virus into rural areas of West Africa. AB - BACKGROUND: In eastern and southern Africa, the human immunodeficiency virus (HIV) epidemic appeared first in urban centres and then spread to rural areas. Its overall prevalence is lower in West Africa, with the highest levels still found in cities. Rural areas are also threatened, however, because of the population's high mobility. We conducted a study in three different communities with contrasting infection levels to understand the epidemiology of HIV infection in rural West Africa. METHOD: A comparative cross-sectional study using a standardized questionnaire and biological tests was conducted among samples in two rural communities of Senegal (Niakhar and Bandafassi, 866 and 952 adults, respectively) and a rural community of Guinea-Bissau (Caio, 1416 adults). We compared the distribution of population characteristics and analysed risk factors for HIV infection in Caio at the individual level. RESULTS: The level of HIV infection was very low in Niakhar (0.3%) and Bandafassi (0.0%), but 10.5% of the adults in Caio were infected, mostly with HIV type 2 (HIV-2). Mobility was very prevalent in all sites. Short-term mobility was found to be a risk factor for HIV infection among men in Caio (adjusted odds ratio (aOR) = 2.06; 95% CI: 1.06 3.99). Women from Caio who reported casual sex in a city during the past 12 months were much more likely to be infected with HIV (aOR = 5.61 95% CI: 1.56 20.15). Short-term mobility was associated with risk behaviours at all sites. CONCLUSIONS: Mobility appears to be a key factor for HIV spread in rural areas of West Africa, because population movement enables the virus to disseminate and also because of the particularly risky behaviours of those who are mobile. More prevention efforts should be directed at migrants from rural areas who travel to cities with substantial levels of HIV infection. PMID- 14559744 TI - Commentary: What can we make of an association between human immunodeficiency virus prevalence and population mobility? PMID- 14559745 TI - The effects of alternative study designs on the power of community randomized trials: evidence from three studies of human immunodeficiency virus prevention in East Africa. AB - BACKGROUND: Randomized intervention trials in which the community is the unit of randomization are increasingly being used to evaluate the impact of public health interventions. In the design of community randomized trials (CRT), the power of the study is likely to be affected by two issues: the matching or stratification of communities, and the number and size of the communities to be randomized. METHODS: Data from three East African community intervention trials, designed to evaluate the impact of interventions to reduce human immunodeficiency virus (HIV) incidence, are used to compare the efficiency of different trial designs. RESULTS: Compared with an unmatched design, stratification reduced the between community variation in the Mwanza trial (from 0.51 to 0.24) and in the Masaka trial (from 0.38 to 0.28). The reduction was smaller in the Rakai trial where the selected communities were more homogeneous (from 0.15 to 0.11). For all trials, individual matching of communities produced estimates of between-community variation similar to those from the stratified designs. The linear association between HIV prevalence and incidence was strong in the Mwanza trial (correlation coefficient R = 0.83) and the Masaka trial (R = 0.83), but weak in the Rakai trial (R = 0.28). Unmatched study designs that use smaller communities tend to increase between-community variation, but reduce the design effect and improve study power. CONCLUSIONS: These empirical data suggest that selection of homogeneous communities, or stratification of communities prior to randomization, may improve the power of CRT. PMID- 14559746 TI - Recent transmission of tuberculosis in Madrid: application of capture-recapture analysis to conventional and molecular epidemiology. AB - BACKGROUND: Population-based studies using a combination of molecular techniques and conventional epidemiological methods have been used to study the dynamics of tuberculosis (TB) transmission but the relative utility of each technique has not yet been established. METHODS: A prospective population-based molecular and epidemiological study of patients diagnosed with TB was conducted in three urban districts of Madrid (Spain) during 1997-1999. Analysis was performed using the capture-recapture method including covariates in which conventional epidemiological data and the information on clustered cases obtained by DNA fingerprinting were regarded as independent and complementary procedures. RESULTS: The estimate obtained by molecular analysis alone, that 31.6% of TB cases were due to recent transmission, was revised to 44.8% (95% CI: 31.4-58.2) using the capture-recapture method. The estimated completeness of the combined databases for identification of recent transmission was 59.2%. Underestimation of the true prevalence of recent transmission was higher with conventional epidemiology than molecular analysis, particularly for patients <35 years old and those with a history of imprisonment. CONCLUSIONS: In this study, use of the capture-recapture technique allowed us to combine epidemiological information obtained by conventional and molecular methods to quantify the number of cases of recently transmitted TB in the community and identify specific populations at high risk of disease. This information is clearly important because such groups are a prime target for improved TB control measures. In the long term, this combination of techniques may contribute significantly to control the spread of TB. PMID- 14559747 TI - Commentary: Can capture-recapture analysis of epidemiological and molecular data help us understand recent tuberculosis transmission? PMID- 14559748 TI - Poliovirus detection in wastewater and stools following an immunization campaign in Havana, Cuba. AB - BACKGROUND: Recent outbreaks of poliomyelitis caused by vaccine-derived virus have raised concerns that vaccine-derived poliovirus may continue to circulate after eradication. In these outbreaks, the virus appears to have replicated for > or =2 years before detection. Early detection is critical for an effective response to these outbreaks. Although acute flaccid paralysis (AFP) surveillance will remain the standard for poliovirus detection, wastewater sampling could be a useful supplement. In this study, we evaluated the sensitivity of wastewater sampling by concurrently collecting stools from children aged < 3 years attending two neighbourhood clinics in Havana, Cuba, and wastewater from the same neighbourhoods. METHODS: Sample collection was begun during the third week after the national immunization campaign, continued weekly through the seventh week, and was repeated during weeks 15 and 19. Virus detection and titration were performed using both cell culture and polymerase chain reaction techniques. RESULTS: Wastewater sampling was found to be at least as sensitive as stool sampling under these conditions. Poliovirus was isolated from children through week 7, suggesting that viral shedding reached undetectable levels between weeks 8 and 14. The last virus-positive wastewater sample was collected during week 15. CONCLUSIONS: Wastewater sampling under the conditions studied can be a sensitive supplement to AFP surveillance. Similar studies under different conditions are needed to determine the role of wastewater sampling in post-eradication surveillance. PMID- 14559749 TI - Estimating the cumulative number of human immunodeficiency virus diagnoses by cross-linking from four different sources. AB - BACKGROUND: We estimated the cumulative number of people diagnosed with human immunodeficiency virus (HIV) infection in a region of Italy by cross-linking data from four surveillance systems and applying capture-recapture methods. METHODS: The study was conducted using data referring to residents of the Veneto Region (population 4.4 million). We cross-linked data from the AIDS Registry (data 1983 2000), the HIV Registry (1988-2000), the Death Registry (1992-1999), and the Hospital-Discharge Registry (1997-2000), using a code based on name, birth date, and sex. A specific software for capture-recapture models (CARE-1) was used to estimate the size of the target population with two different statistical approaches (sample coverage and log-linear models). RESULTS: A total of 2801 people were reported to the AIDS Registry, 6415 to the HIV Registry, 1598 to the Death Registry as HIV/AIDS-related deaths, and 3330 to the Hospital-Discharge Registry with a diagnosis of HIV infection. Overall, 8723 people were present in at least one registry: 4896 people were present in only one registry, 2387 in two registries, 1286 in three registries, and 154 in all four registries. Using the sample coverage approach, we estimated that, since the beginning of the epidemic in Veneto, an estimated 11 281 people (95% CI: 10 981, 11 621) should have been reported to at least one registry; thus the estimated coverage of the four registries was 77.3% (i.e. 8723/11 281). Results obtained applying the log-linear approach were similar, although the fitting of this model was not adequate. CONCLUSIONS: Cross-linking data from four different sources and applying the capture-recapture method can improve the accuracy of the estimates of the dimensions of the HIV epidemic. PMID- 14559750 TI - Predicting incidence of variant Creutzfeldt-Jakob disease from UK dietary exposure to bovine spongiform encephalopathy for the 1940 to 1969 and post-1969 birth cohorts. AB - BACKGROUND: To investigate variant Creutzfeldt-Jakob disease (vCJD) incubation period, transmission barrier, and short-term vCJD predictions for methionine homozygotes in 1940-1969 and post-1969 birth cohorts by use of gender- and age specific exposure intensities to bovine spongiform encephalopathy (BSE), based on consumption of beef mechanically recovered meat (MRM) and head meat. METHODS: Simulation (from vCJD infections generated randomly from gender and age-specific dietary exposure intensities to BSE), constrained to equal the 47 and 64 vCJD onsets before 2001 in 1940-1969 and post-1969 birth cohorts, was used to estimate lognormal (and other) incubation mean and standard deviation which fitted the calendar year distribution of observed vCJD onsets; and to explore exponential decay in susceptibility to infection with age above 15 years. RESULTS: For the post-1969 birth cohort, the best-fitting lognormal incubation period mean of 11 years (SD 1.5 years and 195 infections) was associated with 194 vCJD onsets (64 before 2001, 105 in 2001-2005, and 25 in 2006-2010). About one-fifth of simulated vCJD onsets before 2001 arose from infections in 1990-1996; age and gender of simulated and observed vCJD patients agreed closely. For the 1940-1969 birth cohort, well-fitting lognormal means ranged widely, the marginally best fitting being 26 years (SD 16.5 years and 382 infections; 47 vCJD onsets before 2001, 58 in 2001-2005, and 63 in 2006-2010). An age-dependent susceptibility function was required to match the age distribution of vCJD patients in the 1940-1969 birth cohort. CONCLUSIONS: About three-fifths of predicted vCJD onsets are expected to be in males, and nearly two-thirds of vCJD onsets in 2001-2005 are expected to be in post-1969 birth cohort according to best-fitting predictions. PMID- 14559751 TI - Commentary: Predicting the unpredictable: the future incidence of variant Creutzfeldt-Jakob disease. PMID- 14559753 TI - Commentary: Evaluating the tuberculosis burden in prisoners in Pakistan. PMID- 14559752 TI - Prevalence of and risk factors associated with Mycobacterium tuberculosis infection in prisoners, North West Frontier Province, Pakistan. AB - BACKGROUND: This cross-sectional study was conducted to assess the prevalence of and identify factors associated with latent Mycobacterium tuberculosis (MTB) infection in prisoners of North West Frontier Province (NWFP). METHODS: A stratified random sampling technique was used to select a sample of 425 from a total of 6607 male prisoners aged 18-60 years from the five central prisons of NWFP, Pakistan (Peshawar, Dera Ismail Khan, Haripur, Kohat, and Mardan). The selected inmates were interviewed using a structured pre-tested questionnaire; a Mantoux tuberculin skin test (TST) was also performed. Latent MTB infection status of the prisoner was determined by the size of the induration of the TST in the presence/absence of a previous BCG scar. RESULTS: Overall prevalence of latent MTB infection among prisoners was 48% (204/425). Using multiple logistic regression, a prisoner's age, educational level, smoking status, duration of current incarceration, and average accommodation area of 60 ft(2) or less in prison barracks were found to be statistically significant (P < 0.05) predictors of latent MTB infection. CONCLUSION: The high prevalence of latent MTB infection as assessed by TST in prisoners requires immediate steps be taken to identify and confirm MTB infection, and to treat and counsel those found to be positive in this setting. Efforts to halt MTB transmission in prisons should include: routine screening of prisoners on entry using sputum smear and TST for diagnosis of tuberculosis (TB) and latent MTB infection, respectively. The isolation and treatment of TB positive prisoners and chemo-prophylactic treatment of TST positives, reduction of overcrowding, education regarding the harmful effects of smoking, and intensive monitoring of those serving longer prison terms may help reduce the MTB transmission in this setting and in the community at large. PMID- 14559754 TI - Pipe and cigar smoking and major cardiovascular events, cancer incidence and all cause mortality in middle-aged British men. AB - BACKGROUND: Pipe and cigar smoking are still regarded by many as less hazardous to health than cigarette smoking. METHODS: Prospective study of 7735 men aged 40 59 years drawn from general practices in 24 British towns with mean follow-up of 21.8 years. The outcome measures include major coronary heart disease (CHD) and stroke events, cancer incidence, and deaths from all causes. RESULTS: There were 1133 major CHD events and 440 stroke events, 919 new cancers and 1994 deaths from all causes in the 7121 men with no diagnosed CHD, stroke, diabetes, or cancer at screening. Compared with never smokers, pipe/cigar smokers (primary and secondary combined) showed significantly higher risk of major CHD events (relative risk [RR] = 1.69, 95% CI: 1.32, 2.14) and stroke events (RR = 1.62, 95% CI: 1.08, 2.41) and of cardiovascular, non-cardiovascular, and total mortality (RR = 1.49, 95% CI: 1.13, 1.96, RR = 1.40, 95% CI: 1.08, 1.83 and RR = 1.44, 95% CI: 1.19, 1.74, respectively), after adjustment for lifestyle and biological characteristics. They also showed a significantly higher incidence of smoking related cancers (RR = 2.67, 95% CI: 1.70, 4.26), largely due to lung cancer (RR = 4.35, 95% CI: 2.05, 8.94). Overall, the effects in pipe/cigar smokers were intermediate between never-smokers and light cigarette smokers, although risks for lung cancer were similar to light cigarette smokers. CONCLUSION: Pipe and cigar smoking, whether primary or secondary, carries significant risk of smoking related ill health. PMID- 14559755 TI - Impact of walking upon medical care expenditure in Japan: the Ohsaki Cohort Study. AB - BACKGROUND: Physical activity is expected to reduce medical costs by lowering the risk for a variety of chronic diseases. However, little is known about the actual magnitude of medical cost saved by physical activity. We attempted to quantify the association between time spent walking and medical care costs, based on a 4 year prospective observation of National Health Insurance (NHI) beneficiaries in rural Japan. METHODS: In 27 431 Japanese men and women, aged 40-79 years, who had no functional limitation or conditions interfering with physical activity at the baseline survey in 1994, we ascertained all hospitalizations, outpatient visits, and the costs through computerized linkage with NHI claims history files between January 1995 and December 1998. RESULTS: Medical costs significantly reduced with longer time spent walking. Per capita medical cost was pound 111.80 per month (95% CI: 109.3, 114.2) in those who walked for < or =30 minutes/day, pound 108.10 (95% CI: 105.7, 110.5) in those who walked for 30 minutes-1 hour, and pound 97.30 (95% CI: 95.5, 99.0) in those who walked for > or =1 hour, after multivariate adjustment of potential confounders. CONCLUSIONS: This prospective study in Japan indicated that time spent walking was significantly associated with lower medical costs. PMID- 14559756 TI - Commentary: Can walking lower medical care costs? PMID- 14559757 TI - Women's perception of the benefits of mammography screening: population-based survey in four countries. AB - BACKGROUND: Screening programmes are often actively promoted to achieve high coverage, which may result in unrealistic expectations. We examined women's understanding of the likely benefits of mammography screening. METHODS: Telephone survey of random samples of the female population aged > or =15 years in the US, UK, Italy, and Switzerland using three closed questions on the expected benefits of mammography screening. RESULTS: A total of 5964 women were contacted and 4140 women (69%) participated. Misconceptions were widespread: a majority of women believed that screening prevents or reduces the risk of contracting breast cancer (68%), that screening at least halves breast cancer mortality (62%), and that 10 years of regular screening will prevent 10 or more breast cancer deaths per 1000 women (75%). In multivariate analysis higher number of correct answers was positively associated with higher educational status (odds ratio [OR] = 1.44, 95% CI: 1.25, 1.66) and negatively with having had a mammography in the last 2 years (OR = 0.86, 95% CI: 0.73, 1.01). Compared with US women (reference group) and Swiss women (OR = 0.98, 95% CI: 0.82, 1.18) respondents in Italy (OR = 0.61, 95% CI: 0.50, 0.74) and the UK (OR = 0.73, 95% CI: 0.60, 0.88) gave fewer correct answers. CONCLUSION: In the US and three European countries a high proportion of women overestimated the benefits that can be expected from screening mammography. This finding raises doubts on informed consent procedures within breast cancer screening programmes. PMID- 14559758 TI - Hypothesis: Vitamin A supplementation and childhood mortality: amplification of the non-specific effects of vaccines? AB - Most areas of health research will have accepted data and a dominating interpretation. If the interpretation is not correct, contradictions will accumulate, and it will eventually become clear that the current interpretation is untenable. In this situation, the best hypothesis is the one that accounts for all of the known data as well as the apparent contradictions. The area of vitamin A supplementation and childhood mortality in developing countries is afflicted with many contradictions and there is a need for a new hypothesis. We propose that the effect of vitamin A supplementation may depend on the amplification of non-specific effects of vaccines on childhood mortality. PMID- 14559759 TI - Commentary: A hypothesis concerning vitamin A supplementation, vaccines, and childhood mortality. PMID- 14559761 TI - Commentary: Is there a common background behind growing inequalities in mortality in Western European countries? PMID- 14559760 TI - Widening socioeconomic inequalities in mortality in six Western European countries. AB - OBJECTIVES: During the past decades a widening of the relative gap in death rates between upper and lower socioeconomic groups has been reported for several European countries. Although differential mortality decline for cardiovascular diseases has been suggested as an important contributory factor, it is not known what its quantitative contribution was, and to what extent other causes of death have contributed to the widening gap in total mortality. METHODS: We collected data on mortality by educational level and occupational class among men and women from national longitudinal studies in Finland, Sweden, Norway, Denmark, England/Wales, and Italy (Turin), and analysed age-standardized death rates in two recent time periods (1981-1985 and 1991-1995), both total mortality and by cause of death. For simplicity, we report on inequalities in mortality between two broad socioeconomic groups (high and low educational level, non-manual and manual occupations). RESULTS: Relative inequalities in total mortality have increased in all six countries, but absolute differences in total mortality were fairly stable, with the exception of Finland where an increase occurred. In most countries, mortality from cardiovascular diseases declined proportionally faster in the upper socioeconomic groups. The exception is Italy (Turin) where the reverse occurred. In all countries with the exception of Italy (Turin), changes in cardiovascular disease mortality contributed about half of the widening relative gap for total mortality. Other causes also made important contributions to the widening gap in total mortality. For these causes, widening inequalities were sometimes due to increasing mortality rates in the lower socioeconomic groups. We found rising rates of mortality from lung cancer, breast cancer, respiratory disease, gastrointestinal disease, and injuries among men and/or women in lower socioeconomic groups in several countries. CONCLUSIONS: Reducing socioeconomic inequalities in mortality in Western Europe critically depends upon speeding up mortality declines from cardiovascular diseases in lower socioeconomic groups, and countering mortality increases from several other causes of death in lower socioeconomic groups. PMID- 14559762 TI - Comparison of methods for analysing cluster randomized trials: an example involving a factorial design. AB - BACKGROUND: Studies involving clustering effects are common, but there is little consistency in their analysis. Various analytical methods were compared for a factorial cluster randomized trial (CRT) of two primary care-based interventions designed to increase breast screening attendance. METHODS: Three cluster-level and five individual-level options were compared in respect of log odds ratios of attendance and their standard errors (SE), for the two intervention effects and their interaction. Cluster-level analyses comprised: (C1) unweighted regression of practice log odds; (C2) regression of log odds weighted by their inverse variance; (C3) random-effects meta-regression of log odds with practice as a random effect. Individual-level analyses comprised: (I1) standard logistic regression ignoring clustering; (I2) robust SE; (I3) generalized estimating equations; (I4) random-effects logistic regression; (I5) Bayesian random-effects logistic regression. Adjustments for stratification and baseline variables were investigated. RESULTS: As expected, method I1 was highly anti-conservative. The other, valid, methods exhibited considerable differences in parameter estimates and standard errors, even between the various random-effects methods based on the same statistical model. Method I4 was particularly sensitive to between-cluster variation and was computationally stable only after controlling for baseline uptake. CONCLUSIONS: Commonly used methods for the analysis of CRT can give divergent results. Simulation studies are needed to compare results from different methods in situations typical of cluster trials but when the true model parameters are known. PMID- 14559763 TI - Indoor air pollution from biomass combustion and acute respiratory illness in preschool age children in Zimbabwe. AB - BACKGROUND: Reliance on biomass for cooking and heating exposes many women and young children in developing countries to high levels of air pollution indoors. This study investigated the association between household use of biomass fuels for cooking and acute respiratory infections (ARI) in preschool age children (<5 years) in Zimbabwe. METHODS: Analysis is based on 3559 children age 0-59 months included in the 1999 Zimbabwe Demographic and Health Survey (ZDHS). Children who suffered from cough accompanied by short, rapid breathing during the 2 weeks preceding the survey were defined as having suffered from ARI. Logistic regression was used to estimate the odds of suffering from ARI among children from households using biomass fuels (wood, dung, or straw) relative to children from households using cleaner fuels (liquid petroleum gas [LPG]/natural gas, or electricity), after controlling for potentially confounding factors. RESULTS: About two-thirds (66%) of children lived in households using biomass fuels and 16% suffered from ARI during the 2 weeks preceding the survey interview. After adjusting for child's age, sex, birth order, nutritional status, mother's age at childbirth, education, religion, household living standard, and region of residence, children in households using wood, dung, or straw for cooking were more than twice as likely to have suffered from ARI as children from households using LPG/natural gas or electricity (OR = 2.20; 95% CI: 1.16, 4.19). CONCLUSIONS: Household use of high pollution biomass fuels is associated with ARI in children in Zimbabwe. The relationship needs to be further investigated using more direct measures of smoke exposure and clinical measures of ARI. PMID- 14559764 TI - Acute effect of sulphur dioxide from a power plant on pulmonary function of children, Thailand. AB - BACKGROUND: Epidemiological studies have shown reversible declines of lung function in response to air pollution, but research on the independent effect of short-term exposure to ambient sulphur dioxide (SO2) on pulmonary function is limited. This study evaluated the association of short-term exposure to increased ambient SO2 and daily pulmonary function changes among children with and without asthma. METHODS: The associations of daily exposure to SO2 and particulate matter 10 microm in diameter (PM10) with pulmonary function were examined in 175 asthmatic and non-asthmatic children aged 6-14 years who resided near a coal fired power plant in Thailand. Each child performed daily pulmonary function tests during the 61-day study period. General linear mixed models were used to estimate the association of air pollution and pulmonary function controlling for time, temperature, co-pollutants, and autocorrelation. RESULTS: In the asthmatic children, a daily increase in SO2 was associated with negligible declines in pulmonary function, but a small negative association was found between PM10 and pulmonary function. A 10-microg/m(3) increment was associated with changes in the highest forced vital capacity (FVC) (-6.3 ml, 95% CI: -9.8, -2.8), forced expiratory volume at 1 second (FEV(1)) (-6.0 ml, 95% CI: -9.2, 2.7), peak expiratory flow rate (PEFR) (-18.9 ml.sec(-1), 95% CI: -28.5, -9.3) and forced expiratory flow 25 to 75% of the FVC (FEF(25-75%)) (-3.7 ml.sec(-1), 95% CI: 10.9, 3.5). No consistent associations between air pollution and pulmonary function were found for non-asthmatic children. CONCLUSION: Declines in pulmonary function among asthmatic children were associated with increases in particulate air pollution, rather than with increases in SO2. PMID- 14559765 TI - Relationship between birthweight and blood lipid concentrations in later life: evidence from the existing literature. AB - BACKGROUND: It has been suggested that there is a link between fetal growth and chronic diseases later in life. Several studies have shown a negative association between birthweight and cardiovascular diseases, as well as cardiovascular disease risk factors, such as blood pressure and type 2 diabetes. Far fewer studies have focused on the association between size at birth and blood lipid concentrations. We have conducted a qualitative assessment of the direction and consistency of the relationship between size at birth and blood lipid concentrations to see whether the suggested relationship between intrauterine growth and cardiovascular diseases is mediated by lipid metabolism. METHODS: A literature search covering the period January 1966 to January 2003 was performed using Medline, Embase, and Web of Science. All papers written in English and reporting the relationship between size at birth and lipid levels in humans were assessed. Bibliographies were searched for further publications. RESULTS: From an initial screen of 1198 references, 39 papers were included involving 28 578 individuals. There was no consistent relationship between size at birth and blood lipid levels; the one exception being triglyceride concentration, which showed statistically significant negative or U-shaped, but not positive, relationships with birthweight. CONCLUSION: This review does not strongly support a link between birthweight and blood lipid levels in later life. However, the research in this area is limited and in order to make any definitive conclusions, longitudinal studies with sufficient power, data, and prospective follow-up are needed. PMID- 14559766 TI - Commentary: Developmental origins of raised serum cholesterol. PMID- 14559767 TI - Is the Mediterranean diet relevant to myocardial infarction? Olive oil consumption versus geographical latitude. PMID- 14559768 TI - Can dietary fatty acids affect colon cancer risk? Reply to Leitzmann and Giovannucci. PMID- 14559769 TI - Epidemiologists: clinging to coat-tails or donning them? PMID- 14559771 TI - Sidney Kark's contributions to epidemiology and community medicine. PMID- 14559772 TI - Sostrup statement on low birthweight. PMID- 14559774 TI - The HRT story: lessons for epidemiologists. PMID- 14559775 TI - Fine mapping of the alpha-T catenin gene to a quantitative trait locus on chromosome 10 in late-onset Alzheimer's disease pedigrees. AB - Using plasma amyloid beta protein (Abeta42) levels as an intermediate, quantitative phenotype for late onset Alzheimer's disease (LOAD), we previously obtained significant linkage at approximately 80 cM on chromosome 10. Linkage to the same region was obtained independently in a study of affected LOAD sib-pairs. Together, these two studies provide strong evidence for a novel LOAD locus on chromosome 10 that acts to increase Abeta42. VR22 is a large (1.7 Mb) gene located at 80 cM that encodes alpha-T catenin, which is a binding partner of beta catenin. This makes VR22 an attractive candidate gene because beta catenin interacts with presenilin 1, which has many mutations that elevate Abeta42 and cause early onset familial AD. We identified two intronic VR22 SNPs (4360 and 4783) in strong linkage disequilibrium (LD) that showed highly significant association (P=0.0001 and 0.0006) with plasma Abeta42 in 10 extended LOAD families. This association clearly contributed to the linkage at approximately 80 cM because the lod scores decreased when linkage analysis was performed conditional upon the VR22 association. This association replicated in another independent set of 12 LOAD families (P=0.04 for 4783 and P=0.08 for 4360). Bounding of the association region using multiple SNPs showed VR22 to be the only confirmed gene within the region of association. These findings indicate that VR22 has variant(s) which influence Abeta42 and contribute to the previously reported linkage for plasma Abeta42 in LOAD families. PMID- 14559776 TI - The polyglutamine neurodegenerative protein ataxin-3 binds polyubiquitylated proteins and has ubiquitin protease activity. AB - The ubiquitin-proteasome pathway is critically involved in the pathology of neurodegenerative diseases characterized by protein misfolding and aggregation. Data in the present study suggest that the polyglutamine neurodegenerative disease protein, ataxin-3 (AT3), functions in the ubiquitin-proteasome pathway. AT3 contains an ubiquitin interaction motif (UIM) domain that binds polyubiquitylated proteins with a strong preference for chains containing four or more ubiquitins. Mutating the conserved leucine in the first UIM (L229A) almost totally eliminates binding to polyubiquitin chains while a similar mutation in the second UIM (L249A) also inhibits binding to polyubiquitin chains but to a lesser extent. Both wild-type and pathological AT3 increase cellular levels of a short-lived GFP that is degraded by the ubiquitin-proteasome pathway. AT3 has several properties characteristic of ubiquitin proteases including decreasing polyubiquitylation of 125I-lysozyme by removing ubiquitin from polyubiquitin chains, cleaving a ubiquitin protease substrate, and binding the specific ubiquitin protease inhibitor, ubiquitin-aldehyde. Mutating the predicted catalytic cysteine in AT3 inhibits each of these ubiquitin protease activities. The ability to bind and cleave ubiquitylated proteins is consistent with AT3 playing a role in the ubiquitin-proteasome system. This raises the possibility that pathological AT3, which tends to misfold and aggregate, may be exposed to aggregate-prone misfolded/denatured proteins as part of its normal function. PMID- 14559777 TI - Plectin 5'-transcript diversity: short alternative sequences determine stability of gene products, initiation of translation and subcellular localization of isoforms. AB - Plectin is a large cytoskeletal linker protein expressed as several different isoforms from a highly complex gene. This transcript diversity is mainly caused by short 5'-sequences contained in alternative first exons. To elucidate the influence of these sequence differences and to determine potential differential functionality of the resulting protein forms, we conducted a systematic investigation of plectin isoforms on transcript and protein levels. Isoform expression was highly dependent on the different 5' ends, largely due to effects of the 5'-untranslated regions. Initiation of translation downstream of the expected start site led to loss of actin- and integrin beta4-binding in some isoforms. The small alternative N-terminal sequences (5-180 residues) profoundly affected the subcelluar localization of this >500 kDa protein. Specifically, plectin 1f was concentrated at focal adhesion contacts and plectin 1b was exclusively targeted to mitochondria, providing a connection of these organelles to intermediate filaments. Thus, with plectin as a model, we demonstrate a role for 5'-untranslated regions and alternative 5'-splicing as an important regulatory mechanism of protein expression and protein function. PMID- 14559778 TI - Bioinformatics analysis of experimentally determined protein complexes in the yeast Saccharomyces cerevisiae. AB - Many important cellular functions are implemented by protein complexes that act as sophisticated molecular machines of varying size and temporal stability. Here we demonstrate quantitatively that protein complexes in the yeast Saccharomyces cerevisiae are comprised of a core in which subunits are highly coexpressed, display the same deletion phenotype (essential or nonessential), and share identical functional classification and cellular localization. This core is surrounded by a functionally mixed group of proteins, which likely represent short-lived or spurious attachments. The results allow us to define the deletion phenotype and cellular task of most known complexes, and to identify with high confidence the biochemical role of hundreds of proteins with yet unassigned functionality. PMID- 14559779 TI - Functionality of system components: conservation of protein function in protein feature space. AB - Many protein features useful for prediction of protein function can be predicted from sequence, including posttranslational modifications, subcellular localization, and physical/chemical properties. We show here that such protein features are more conserved among orthologs than paralogs, indicating they are crucial for protein function and thus subject to selective pressure. This means that a function prediction method based on sequence-derived features may be able to discriminate between proteins with different function even when they have highly similar structure. Also, such a method is likely to perform well on organisms other than the one on which it was trained. We evaluate the performance of such a method, ProtFun, which relies on protein features as its sole input, and show that the method gives similar performance for most eukaryotes and performs much better than anticipated on archaea and bacteria. From this analysis, we conclude that for the posttranslational modifications studied, both the cellular use and the sequence motifs are conserved within Eukarya. PMID- 14559780 TI - Investigations into the analysis and modeling of the TNF alpha-mediated NF-kappa B-signaling pathway. AB - In this study, we propose a system-theoretic approach to the analysis and quantitative modeling of the TNFalpha-mediated NF-kappaB-signaling pathway. Tumor necrosis factor alpha (TNFalpha) is a potent proinflammatory cytokine that plays an important role in immunity and inflammation, in the control of cell proliferation, differentiation, and apoptosis. To date, there have been numerous approaches to model cellular dynamics. The most prominent uses ordinary differential equations (ODEs) to describe biochemical reactions. This approach can provide us with mathematically well-founded and tractable interpretations regarding pathways, especially those best described by enzyme reactions. This work first introduces a graphical method to intuitively represent the TNFalpha mediated NF-kappaB-signaling pathway and then utilizes ODEs to quantitatively model the pathway. The simulation study shows qualitative validation of the proposed model compared with experimental results for this pathway. The proposed system-theoretic approach is expected to be further applicable to predict the signaling behavior of NF-kappaB in a quantitative manner for any variation of the ligand, TNFalpha. PMID- 14559781 TI - In silico atomic tracing by substrate-product relationships in Escherichia coli intermediary metabolism. AB - We present a software system that computationally reproduces biochemical radioisotope-tracer experiments. It consists of three main components: A mapping database of substrate-product atomic correspondents derived from known reaction formulas, a tracing engine that can compute all pathways between two given compounds by using the mapping database, and a graphical user interface. As the system can facilitate the display of all possible pathways between any two compounds and the tracing of every single carbon, nitrogen, or sulfur atom in the metabolism, it complements and bridges other metabolic databases and simulations on fixed models. PMID- 14559782 TI - Signal processing and flagellar motor switching during phototaxis of Halobacterium salinarum. AB - Prokaryotic taxis, the active search of motile cells for the best environmental conditions, is one of the paradigms for signal transduction. The search algorithm implemented by the cellular biochemistry modulates the probability of switching the rotational direction of the flagellar motor, a nanomachine that propels prokaryotic cells. On the basis of the well-known biochemical mechanisms of chemotaxis in Escherichia coli, kinetic modeling of the events leading from chemoreceptor activation by ligand binding to the motility response has been performed with great success. In contrast to Escherichia coli, Halobacterium salinarum, in addition, responds to visible light, which is sensed through specific photoreceptors of different wavelength sensitivity (phototaxis). Light stimuli of defined intensity and time course can be controlled precisely, which facilitates input-output measurements used for system analysis of the molecular network connecting the sensory receptors to the flagellar motor switch. Here, we analyze the response of halobacterial cells to single and double-pulse light stimuli and present the first kinetic model for prokaryotic cells that couples the signal-transduction pathway with the flagellar motor switch. Modeling based on experimental data supports the current biochemical model of halobacterial phototaxis. Moreover, the simulations demonstrate that motor switching occurs through subsequent rate-limiting steps, which are both under sensory control, suggesting that two signals may be involved in halobacterial phototaxis. PMID- 14559783 TI - Parameter estimation in biochemical pathways: a comparison of global optimization methods. AB - Here we address the problem of parameter estimation (inverse problem) of nonlinear dynamic biochemical pathways. This problem is stated as a nonlinear programming (NLP) problem subject to nonlinear differential-algebraic constraints. These problems are known to be frequently ill-conditioned and multimodal. Thus, traditional (gradient-based) local optimization methods fail to arrive at satisfactory solutions. To surmount this limitation, the use of several state-of-the-art deterministic and stochastic global optimization methods is explored. A case study considering the estimation of 36 parameters of a nonlinear biochemical dynamic model is taken as a benchmark. Only a certain type of stochastic algorithm, evolution strategies (ES), is able to solve this problem successfully. Although these stochastic methods cannot guarantee global optimality with certainty, their robustness, plus the fact that in inverse problems they have a known lower bound for the cost function, make them the best available candidates. PMID- 14559785 TI - Extracellular proteolysis and cancer: meeting summary and future directions. AB - The contribution of extracellular proteolysis to processes of tumor invasion and metastasis has been recognized for more than 20 years. However, after millions of dollars and untold hours of investment in preclinical research and the development of broad range inhibitors of MMPs, clinical trials of late-stage tumor patients show no indication that this approach will be successful. In the basic science arena, there have been stunning advancements that illustrate novel biological activities for proteases and that suggest they are key regulators of many physiological and pathological processes. The Proteases and Cancer: Biology and Therapeutics Workshop (held in Bethesda, MD, November 20-22, 2002) was organized by the Division of Cancer Biology, National Cancer Institute (NCI) to identify research areas and directions that will accelerate understanding protease biology and enhance clinical translation. The overall consensus was that protease biology represents fertile ground for advances that will be clinically useful but perhaps not for the reasons or purpose originally thought. Protease related technologies show particular promise for the detection, prognosis, and prevention of cancer, and for therapeutic purposes in defined situations. Promising areas for further research are identified, and specific recommendations for the development of a consortium to coordinate the efforts of the protease community are made. PMID- 14559784 TI - Reconciling gene expression data with known genome-scale regulatory network structures. AB - The availability of genome-scale gene expression data sets has initiated the development of methods that use this data to infer transcriptional regulatory networks. Alternatively, such regulatory network structures can be reconstructed based on annotated genome information, well-curated databases, and primary research literature. As a first step toward reconciling the two approaches, we examine the consistency between known genome-wide regulatory network structures and extensive gene expression data collections in Escherichia coli and Saccharomyces cerevisiae. By decomposing the regulatory network into a set of basic network elements, we can compute the local consistency of each instance of a particular type of network element. We find that the consistency of network elements is influenced by both structural features of the network such as the number of regulators acting on a target gene and by the functional classes of the genes involved in a particular element. Taken together, the approach presented allows us to define regulatory network subcomponents with a high degree of consistency between the network structure and gene expression data. The results suggest that targeted gene expression profiling data can be used to refine and expand particular subcomponents of known regulatory networks that are sufficiently decoupled from the rest of the network. PMID- 14559786 TI - Double RNA interference of DNMT3b and DNMT1 enhances DNA demethylation and gene reactivation. AB - Small interfering RNAs (siRNAs) are newly identified molecules shown to silence genes via targeted mRNA degradation. In this study, we used specific siRNAs as a tool to probe the relationship between two DNA methyltransferase genes, DNMT3b and DNMT1, in the maintenance of DNA methylation patterns in the genome. Levels of DNMT3b or DNMT1 mRNAs and proteins were markedly decreased (up to 80%) on transfecting these siRNAs into the ovarian cancer cell line CP70. The resulting RNA interference showed differential effects on DNA demethylation and gene reactivation in the treated cells. The DNMT1 siRNA treatment led to a partial removal of DNA methylation from three inactive promoter CpG islands, TWIST, RASSF1A, and HIN-1, and restored the expression of these genes. This epigenetic alteration appeared less effective in cells transfected with DNMT3b siRNA. However, the combined treatment of DNMT3b and DNMT1 siRNAs greatly enhanced this demethylation effect, producing 7-15-fold increases in their expression. We also used a microarray approach to examine this RNA interference on 8640 CpG island loci in CP70 cells. The combined siRNA treatment had a greater demethylation effect on 241 methylated loci and selected repetitive sequences than that of the single treatment. Our data thus suggest that whereas DNMT1 plays a key role in methylation maintenance, DNMT3b may act as an accessory to support the function in CP70 cells. This study also shows that siRNA is a powerful tool for interrogating the mechanisms of DNA methylation in normal and pathological genomes. PMID- 14559788 TI - In vivo opossum xenograft model for cancer research. AB - The inadequacies of athymic mice for research with grafted tumors are well known. Monodelphis domestica, the laboratory opossum, has the potential to complement the rodent model because of the immunoincompetent window period during early marsupial development. We injected pups of different ages with mouse melanoma cells and human melanoma, colon, and prostate cancer cells. The results showed that the xenogeneic tumors can grow and metastasize. We also obtained positive results by heterotopically injecting juveniles with mouse melanoma cells. These results establish Monodelphis as a natural mammalian model to study the cascade of interactions between xenografted cells and the host system. PMID- 14559787 TI - Involvement of the FGF18 gene in colorectal carcinogenesis, as a novel downstream target of the beta-catenin/T-cell factor complex. AB - To search for potential molecular targets for development of novel anticancer drugs, we have been analyzing expression profiles of clinical samples from cancer patients, using a genome-wide cDNA microarray. In experiments with colon cancer cells, the gene encoding fibroblast growth factor 18 (FGF18) was among those that showed elevated expression. The promoter region of this gene was found to contain putative Tcf4-binding motifs; moreover a reporter-gene assay using luciferase activity as a marker and an electromobility shift assay indicated that FGF18 is a downstream transcription target in the beta-catenin/Tcf4 pathway. We showed that exogenous FGF18 promoted growth of NIH3T3 cells in an autocrine manner and that transfection of FGF18 short interfering RNAs suppressed growth of colon cancer cells in culture. Our results indicate that FGF18 is activated in colon cancers as a direct downstream target of the Wnt signaling pathway and that it might represent a marker for early diagnosis and a molecular target for treatment of this life-threatening tumor. PMID- 14559789 TI - Different susceptibility of human mesothelial cells to polyomavirus infection and malignant transformation. AB - SV40 has been associated with mesothelioma development. The possible role of the closely related human polyomaviruses JC virus (JCV) and BK virus (BKV) in mesothelioma remained unclear. We found that JCV did not infect human mesothelial cells. BKV and SV40 infected mesothelial cells, expressed viral oncoproteins, and caused similar alterations of key cell regulatory genes. BKV replicated faster than SV40 and caused mesothelial cell lysis, not cellular transformation. SV40 did not lyse mesothelial cells and caused a high rate of transformation. These findings provide a rationale for the observation that SV40 is found in mesothelioma, rather than the ubiquitous human JCV and BKV. PMID- 14559791 TI - Menin, a tumor suppressor, represses JunD-mediated transcriptional activity by association with an mSin3A-histone deacetylase complex. AB - Menin, a gene product of multiple endocrine neoplasia type I (MEN1), is known to act as a tumor suppressor to repress JunD transcription factor. However, the mechanism by which Menin represses JunD transcriptional activity was still unclear. In this study, we found that Menin is a corepressor against JunD transcriptional activity via recruitment of histone deacetylases in an mSin3A dependent manner. The amino acid search revealed that central domain of Menin includes a alpha-helical mSin3-interacting domain [SID (371-387)]. The SID mutation of Menin (L381P/A385P) abolished the interaction between mSin3A and paired amphipathic helix 2 domain of Menin and reduced its ability to repress JunD transcriptional activity, implicating that SID of Menin is important for recruiting an mSin3A-histone deacetylase complex to repress JunD transcriptional activity. PMID- 14559790 TI - Predominant role of hypoxia-inducible transcription factor (Hif)-1alpha versus Hif-2alpha in regulation of the transcriptional response to hypoxia. AB - Tumor hypoxia induces the up-regulation of a gene program associated with angiogenesis, glycolysis, adaptation to pH, and apoptosis via the hypoxia inducible transcription factors (Hifs) 1 and 2. Disruption of this pathway has been proposed as a cancer therapy. Here, we use short interfering RNAs to compare specific inactivation of Hif-1alpha or Hif-2alpha and show markedly different cell type-specific effects on gene expression and cell migration. Remarkably, among a panel of hypoxia-inducible genes, responses were critically dependent on Hif-1 alpha but not Hif-2 alpha in both endothelial and breast cancer cells but critically dependent on Hif-2 alpha in renal carcinoma cells. PMID- 14559792 TI - p53 function is required for hormone-mediated protection of mouse mammary tumorigenesis. AB - Limited hormonal stimulation of the mammary gland during a critical window in postpubertal development imparts a long-lasting protective effect against breast cancer in humans and in rodent models. The hormonal stimulation can be achieved by full-term pregnancy or low doses of estradiol-17beta and progesterone administered for 21 days. The mechanism(s) behind this effect of hormones is not understood at the molecular level. The experiments reported here demonstrate that the absence of p53 tumor suppressor gene function abrogates the protective effect of hormones against carcinogen-induced mammary carcinogenesis in BALB/c mice. This is the first identification of a specific gene product that mediates the protective effect of hormones. Additionally, the experiments highlight the usefulness of transgenic mouse models in the testing of hypotheses derived from the classic rat mammary models. PMID- 14559793 TI - Inhibition of the translocated c-myc in Burkitt's lymphoma by a PNA complementary to the E mu enhancer. AB - In Burkitt's Lymphoma there is an up-regulation of the c-myc oncogene caused by its translocation from chromosome 8 to chromosome 14, often close to the E mu enhancer of the immunoglobulin heavy chain locus (IgH). In Burkitt's Lymphoma cells, a peptide nucleic acid complementary for a specific unique E mu intronic sequence selectively blocked the expression of the c-myc oncogene under E mu control but not of other c-myc alleles. This Peptide Nucleic Acid also inhibited IgM expression in B cells. The finding that PNAs specific for a regulatory noncoding sequence can block gene expression has important conceptual and practical implications. PMID- 14559794 TI - Resistance to senescence induction and telomere shortening by a G-quadruplex ligand inhibitor of telomerase. AB - The molecular mechanisms induced by G-quadruplex ligands to trigger senescence in mammalian cells are still unknown, although the critical role of telomerase is highly suspected. JFA2 cells selected for resistance to senescence induced by the G-quadruplex ligand 12459 presented an overexpression of hTERT transcript that correlated to a functional increase in telomerase activity and telomere length. Consistently, treatment with 12459 failed to trigger senescence and telomere shortening in JFA2 cells. Resistant cells also presented cross-resistance for senescence induction to telomestatin, another G-quadruplex ligand from a different series, but not to other anticancer agents, indicating the selectivity of the resistance mechanism. We, thus, provide evidence that telomerase activity and telomere length are key cellular determinants of the resistance to G quadruplex ligands. PMID- 14559795 TI - Polypyrimidine tract-binding protein down-regulates fibroblast growth factor receptor 1 alpha-exon inclusion. AB - Exclusion of the alpha-exon by alternative RNA splicing of the fibroblast growth factor receptor 1 (FGFR1) primary transcript leads to the production of FGFR1beta. Glial cell transformation is associated with a progressive increase in FGFR1beta expression that coincides with a dramatic increase in the expression of the splicing factor polypyrimidine tract-binding protein (PTB). Cell-specific overexpression of PTB increased alpha-exon skipping, and a reduction in PTB increased alpha-exon inclusion. Targeted disruption of PTB interaction with FGFR1 precursor RNA in vivo by an antisense oligonucleotide also increased alpha-exon inclusion. These results suggest that PTB plays a direct role in alpha-exon splicing. PMID- 14559796 TI - High epithelial and stromal genetic instability of chromosome 17 in ulcerative colitis-associated carcinogenesis. AB - To define the relative frequencies of genetic instability in stromal and epithelial compartments during ulcerative colitis (UC)-associated tumorigenesis, samples from laser-captured microdissection were assessed for microsatellite instability and loss of heterozygosity in regenerative tissue, dysplasia, and carcinomas in long-standing UC cases. Five National Cancer Institute-recommended standard markers and four markers located close to p53 and BRCA1 genes in chromosome 17 were tested, and p53 gene sequencing was also carried out. Although chromosome 17-MSI and -loss of heterozygosity in epithelium correlated with histological progression, in stroma they showed a consistently high frequency throughout the different stages, indicating a distinct carcinogenesis pathway of UC. The rates for standard markers were lower in both epithelium and stroma. PMID- 14559797 TI - Human recombinant erythropoietin significantly improves tumor oxygenation independent of its effects on hemoglobin. AB - Tumor oxygenation is known to be an important predictive/prognostic marker in a variety of tumors, including cervix, head/neck, sarcoma, non-small cell of the lung, and breast. Tumor oxygenation is influenced by many interactions, including oxygen delivery (angiogenesis, permeability, and HgB) and consumption (metabolic and growth rates). This study randomized 30 nonanemic, female Fischer 344 rats into three treatment arms to examine the effects of recombinant human erythropoietin (EPO) on R3230 rodent mammary carcinoma oxygenation. The three treatment arms were: (a) placebo; (b) EPO after tumor implantation (2000 units/kg/SQdose, M/W/F for six doses); and (c) EPO before tumor implantation (2000 units/kg/SQdose, M/W/F for six doses). Tumors were implanted in the hindflank, and in vivo oxygenation was measured at day 22 after implantation using the Oxylite system (Oxford Optronix, Oxford, England). An average of 180 measurements/animal were performed. On day 22, median tumor volume was 399 mm(3) (range: 65-950 mm(3)), and no differences in tumor volume were seen between treatment arms. Mean hematocrit was equal between arms at therapy initiation but were significantly higher for both arms receiving EPO at day 22 (placebo versus Arm B versus Arm C; Wilcoxon P = 0.052). EPO-treated tumors had significantly less hypoxic measurements when compared with either the placebo or those receiving EPO before implantation. These data confirm that tumor oxygenation in nonanemic individuals may be improved through the administration of EPO, and this improvement appears to be independent of HgB effects. PMID- 14559798 TI - DNA alterations in human aberrant crypt foci and colon cancers by random primed polymerase chain reaction. AB - Colon cancers are the result of the accumulation of multiple genetic alterations. To evaluate the role genomic instability plays during tumor development, we compared DNA fingerprints of 44 aberrant crypt foci (ACF; the earliest identified neoplastic lesion in the colon), 23 cancers, and normal crypts generated by random primers with PCR. The PCR products, separated by PAGE and viewed after silver staining, demonstrate altered fingerprints for 23.3% of the ACF and 95.7% of the cancers. In this first study of human ACF with this approach, the finding of altered DNA fingerprints in these microscopic lesions suggests that genomic instability can occur very early in human colon tumorigenesis. PMID- 14559799 TI - Ferredoxin reductase: pharmacogenomic assessment in colorectal cancer. AB - Ferredoxin reductase (FDXR) is a putative contributor to TP53-mediated apoptosis from 5-fluorouracil chemotherapy through the generation of oxidative stress. With TaqMan real-time quantitative reverse transcription-PCR, this study established a significant difference in FDXR relative RNA expression level between tumor (median, 212.9 units) and normal tissues (median, 113.8 units) from 51 colorectal cancer patients (P < 0.001). Seven single nucleotide polymorphisms (SNPs) in the FDXR gene were discovered, with no significant difference in variant allele frequency between colon tumor and normal tissues (P > 0.05), and the common haplotypes for FDXR were not different between colon tumor and normal samples. No correlation was observed between FDXR genotype and RNA expression implying that the polymorphisms described in this study are not regulating FDXR expression in colon cancer. This genomic characterization provides the foundation for pharmacogenetic analysis of the impact of FDXR on chemotherapy for colorectal cancer. PMID- 14559800 TI - Blockade of Hsp27 overcomes Bortezomib/proteasome inhibitor PS-341 resistance in lymphoma cells. AB - Bortezomib (PS-341), a selective inhibitor of proteasome, induces apoptosis in various tumor cells, but its mechanism of action is unclear. Treatment with PS 341 induces apoptosis in SUDHL6 (DHL6), but not SUDHL4 (DHL4), lymphoma cells. Microarray analysis shows high RNA levels of heat shock protein-27 (Hsp27) in DHL4 versus DHL6 cells, which correlates with Hsp27 protein expression. Blocking Hsp27 using an antisense strategy restores the apoptotic response to PS-341 in DHL4 cells; conversely, ectopic expression of wild-type Hsp27 renders PS-341 sensitive DHL6 cells resistant to PS-341. These findings provide the first evidence that Hsp27 confers PS-341 resistance. PMID- 14559801 TI - Differential distribution of DNA methylation within the RASSF1A CpG island in breast cancer. AB - Aberrant DNA methylation of promoter CpG islands is associated with transcriptionally repressive heterochromatin in neoplasia. The dynamics of this epigenetic process in mediating the transition from an active to an inactive state of transcription remains to be elucidated, however. Here, we used the methylation-specific oligonucleotide microarray to map the methylation patterns of a CpG island, located within the promoter and the first exon regions of RASSF1A, in normal breast tissue controls, primary tumors, and breast cancer cell lines. Oligonucleotide pairs, spaced along the CpG island region, were designed to discriminate between methylated and unmethylated alleles of selected sites. The methylation-specific oligonucleotide data indicate that the majority of test samples show widespread methylation in the first exon of RASSF1A. In contrast, the promoter area was usually undermethylated in normal controls and in 32% of the primary tumors tested, whereas the rest of the primary tumors and breast cancer cell lines showed various degrees of methylation in the region. Methylation profiling of individual tumors further suggest that DNA methylation progressively spreads from the first exon into the promoter area of this gene. Functional analysis indicates that increased density of RASSF1A promoter methylation is associated with altered chromatin, marked by a depletion of acetylated histones and methylated histone 3-lysine 4 and an enrichment of methylated histone 3-lysine 9 in the studied area. The combination of these epigenetic modifications may engender a stable silencing of the gene in breast cancer cells. Thus, this study underscores the importance of detailed mapping of methylation patterns within a CpG island locus that may provide insights into the progressive nature of aberrant DNA methylation and its relationship with transcriptional silencing during the neoplastic process. PMID- 14559802 TI - Effects of oligonucleotide N3'-->P5' thio-phosphoramidate (GRN163) targeting telomerase RNA in human multiple myeloma cells. AB - Telomeres are specialized nucleoprotein complexes that protect against fusion and degradation of linear chromosomes. Critical shortening of telomeres leads to irreversible cessation of cell division, whereas telomerase elongates telomere sequences to compensate for losses that occur with each round of DNA replication. Continued proliferation of tumor cells requires this enzyme to maintain chromosomal stability and to counteract the cellular mitotic clock. In this study, we evaluated the effect of oligonucleotide N3'-->P5' thio-phosphoramidate (NP), which targets template RNA component, in human multiple myeloma (MM) cell lines and patient MM cells. Fluorescein staining at 24 h confirmed NP uptake in 84.7 and 86.1% of MM.1S cells and MM patient cells, respectively, without any transfection enhancer. High transfection efficiency was observed into both CD138(+) and CD138(-) MM patient cells. Match NP (7S), but not mismatch NP (30S), inhibited telomerase activity in MM.1S cells, U266 cells, and RPMI 8226 cells, as well as in patient MM cells. Moreover, 7S inhibited cytokine-induced telomerase activity in MM.1S cells. 7S treatment-induced progressive telomere shortening was associated with growth inhibition and cell death in MM.1S cells with short telomeres (2.5 kb), but not in U266 cells with long telomeres (9.0 kb), at 56 days of culture. Progressive telomere shortening leading to growth inhibition and cell death in MM.1S cells was associated with up-regulation of p21 and phosphorylation of p53 (Ser-15). These studies, therefore, identify the molecular sequelae of NP oligonucleotide (GRN163) against human telomerase RNA component as a telomerase inhibitor and provide the rationale for the development of telomerase-targeted therapies to improve patient outcome in MM. PMID- 14559803 TI - Coordinate expression of Cdc25B and ER-alpha is frequent in low-grade endometrioid endometrial carcinoma but uncommon in high-grade endometrioid and nonendometrioid carcinomas. AB - Overexpression of cdc25B, an important cell cycle regulator, has been shown to result in mammary gland hyperplasia in transgenic mice and to increase steroid hormone responsiveness as a direct coactivator of the estrogen receptor (ER). We investigated the potential role of cdc25B in the pathogenesis of endometrial carcinomas in conjunction with ER-alpha. We examined the expression of cdc25B and phosphorylated ER-alpha in 4 archived human specimens of normal endometrium; 7 endometrial hyperplasia with or without atypia; 32 endometrioid endometrial carcinoma (EEC), including 20 low-grade (grade 1) and 12 high-grade (grade 2 or 3) tumors; and 18 endometrial cancers with aggressive histological subtypes (uterine papillary serous carcinoma and clear cell carcinoma, UPSC/CCC) by immunohistochemistry with monoclonal antibodies. Expression of cdc25B and phosphorylated ER-alpha was increased in endometrial hyperplasia and atypical hyperplasia compared with normal secretory endometrium. Ninety percent (18 of 20) of the low-grade EEC expressed cdc25B at a high level, whereas only 42% (5 of 12) of the high-grade EEC did so (chi(2) = 8.7; P < 0.01). Sixty-five percent (13 of 20) of the low-grade EEC expressed phosphorylated ER-alpha at high levels, but only 17% (2 of 12) of high-grade EEC did so (chi(2) = 7.0; P < 0.01). Coordinate high-level expression of phosphorylated ER-alpha and cdc25B occurred in 65% (13 of 20) of low-grade EEC but in only 17% (2 of 12) of the high-grade EEC (chi(2) = 7.0; P < 0.01). In the UPSC/CCC tumors, only 22% (4 of 18) of the tumors expressed phosphorylated ER-alpha at high-levels. However, 83% (15 of 18) of these carcinomas showed high expression of cdc25B (chi(2) = 13.5; P < 0.01). The majority of the UPSC/CCC (15 of 18) did not show coordinate high expression of phosphorylated ER-alpha and cdc25B. Our findings show that in endometrial hyperplasia and low-grade EEC, coordinate increase in cdc25B and phosphorylated ER-alpha occurs. However, in UPSC/CCC, cdc25B is highly expressed without coordinate increase in phosphorylated ER-alpha. Cdc25B may play important roles in the development and progression of EEC and UPSC/CCC by different mechanisms. PMID- 14559804 TI - Cytogenetic and morphologic typing of 58 papillary renal cell carcinomas: evidence for a cytogenetic evolution of type 2 from type 1 tumors. AB - We evaluated clinical characteristics, patient outcome (mean follow-up, 47 months), and cytogenetic abnormalities in the largest as yet reported cytogenetic series of 47 primary and 11 secondary papillary renal cell carcinomas for differences between the recently proposed type 1 and type 2 subtypes. Secondary tumors were more often of type 2 morphology (P = 0.02), whereas primary type 2 tumors were associated with higher clinical stage (P = 0.001) and worse patient outcome (P = 0.02). Although both subtypes had at least one of the primary chromosomal gains at 17q, 7, and 16q, type 2 tumors had moderately lower frequencies of primary gains at 17p (61 versus 94%; P = 0.007) and 17q (72 versus 97%; P = 0.02). On the other hand, type 2 tumors overall had more chromosomal alterations than type 1 tumors (P = 0.01), particularly gains of 1q (28 versus 3%; P = 0.02) and losses of 8p (33 versus 0%; P = 0.001), 11 (28 versus 3%; P = 0.02), and 18 (44 versus 9%; P = 0.01). Hierarchical clustering suggested cytogenetic patterns common but not restricted to type 2 morphology, one characterized by multiple additional gains, and another predominantly showing additional losses. These findings provide genetic evidence that type 1 and type 2 tumors arise from common cytogenetic pathways and that type 2 tumors evolve from type 1 tumors. Independently of type, losses of 9p were statistically correlated with advanced disease (P = 0.0008) and may serve as a potential adverse prognostic marker in papillary renal cell carcinomas. PMID- 14559805 TI - Relationship of Ras association domain family 1 methylation and K-ras mutation in primary non-small cell lung cancer. AB - Recently, several groups have reported that Ras association domain family 1 (RASSF1A) interacts with Ras and mediates Ras-dependent apoptosis. However, the mechanism by which RASSF1A plays a role as a tumor suppressor in human cancer is unclear. In this study, we investigated the relationship between the RASSF1A methylation and K-ras mutation and their effects on patient's survival in 242 primary non-small cell lung cancers (NSCLCs) to understand the role of RASSF1A in Ras-mediated oncogenic transformation. RASSF1A methylation was not found to be associated with the K-ras mutation in NSCLCs (P = 0.37). For patients with stage I adenocarcinoma, those with RASSF1A methylation and K-ras mutation had a poorer prognosis than those with either RASSF1A methylation or K-ras mutation (P = 0.001). In stage II-III adenocarcinoma patients, the median survival of those with RASSF1A methylation and K-ras mutation was 9 months, and this was poorer than that of those with either RASSF1A methylation or K-ras mutation (P = 0.001). The hazard of failure for those with RASSF1A methylation and K-ras mutation was approximately 2.94 times higher compared with that of those with neither K-ras mutation nor RASSF1A methylation (95% confidence interval = 1.67-9.42; P = 0.01). Our results suggest that RASSF1A methylation and K-ras mutation are not mutually exclusive in NSCLC. In addition, RASSF1A methylation, in combination with K-ras mutation, may have an adverse synergistic effect on patient's survival in NSCLCs. PMID- 14559806 TI - An evolutionary model of carcinogenesis. AB - A quantitative model of carcinogenesis based on methods from population biology and game theory demonstrates normal cells in vivo occupy a ridge-shaped maximum in a well-defined tissue fitness landscape, a novel configuration that allows cooperative coexistence of multiple cellular populations. This state, although necessary for development of functioning multicellular organisms, is subject to invasion by fitter, mutant phenotypes permitting somatic evolution of cancer. The model demonstrates carcinogenesis is an emergent phenomenon requiring a sequence of evolutionary steps as cellular proliferation follows successful adaptation to varying environmental constraints. In the initial development of preneoplastic lesions, cellular proliferation is controlled exclusively by interactions with other cells, the extracellular matrix, and soluble or insoluble growth factors so that gain of function mutations in oncogenes, loss of function mutations in tumor suppressor genes, and disruption of normal senescence pathways will permit clonal expansion. This provides explicit selection mechanisms for the mutations depicted in the classical Fearon-Vogelstein model of colorectal carcinogenesis. The model demonstrates neoplastic cellular proliferation can also be promoted by alterations in the somatic landscape that reduce inhibitory signals produced by the normal cells and extracellular matrix. This is consistent with experimental evidence for a strong microenvironmental influence in tumorigenesis independent of genomic changes in the neoplastic populations. However, we find that these changes alone produce only self-limited neoplastic growth because cellular crowding alters system dynamics so that proliferation is dependent on substrate availability. Consequent cellular competition for critical nutrients results in Darwinian selection pressures favoring phenotypes that increase substrate delivery (e.g., angiogenesis) or uptake (e.g., amplify membrane transporters). These previously unknown substrate dynamics in the later stages of carcinogenesis provide a mechanism for evolution of cellular properties typically found in invasive cancers including the angiogenic and glycolytic phenotypes. PMID- 14559807 TI - BRCA1 functions as a differential modulator of chemotherapy-induced apoptosis. AB - We have evaluated the role played by BRCA1 in mediating the phenotypic response to a range of chemotherapeutic agents commonly used in cancer treatment. Here we provide evidence that BRCA1 functions as a differential mediator of chemotherapy induced apoptosis. Specifically, we demonstrate that BRCA1 mediates sensitivity to apoptosis induced by antimicrotubule agents but conversely induces resistance to DNA-damaging agents. These data are supported by a variety of experimental models including cells with inducible expression of BRCA1, siRNA-mediated inactivation of endogenous BRCA1, and reconstitution of BRCA1-deficient cells with wild-type BRCA1. Most notably we demonstrate that BRCA1 induces a 10-1000 fold increase in resistance to a range of DNA-damaging agents, in particular those that give rise to double-strand breaks such as etoposide or bleomycin. In contrast, BRCA1 induces a >1000-fold increase in sensitivity to the spindle poisons, paclitaxel and vinorelbine. Fluorescence-activated cell sorter analysis demonstrated that BRCA1 mediates G(2)/M arrest in response to both antimicrotubule and DNA-damaging agents. However, poly(ADP-ribose) polymerase and caspase-3 cleavage assays indicate that the differential effect mediated by BRCA1 in response to these agents occurs through the inhibition or induction of apoptosis. Therefore, our data suggest that BRCA1 acts as a differential modulator of apoptosis depending on the nature of the cellular insult. PMID- 14559808 TI - Adenovirus type 5 E1A sensitizes hepatocellular carcinoma cells to gemcitabine. AB - Hepatocellular carcinoma (HCC) is resistant to conventional chemotherapy. A few clinical trials have shown that the cytidine analogue gemcitabine appears to have antitumor activity for HCC, but the overall survival times remain to be improved. In this study, we examined the synergistic effect of adenovirus type 5 E1A (E1A) and gemcitabine on HCC and found that E1A sensitized J5, J7, Huh7, and HepG2 HCC cells to gemcitabine. To further study the E1A-mediated chemosensitization, we established stable cell lines that expressed the E1A gene and then examined whether E1A could have proapoptotic activity while expressed in HCC cells. Our results clearly showed that E1A sensitized HCC cells to gemcitabine through induction of apoptosis. To study the underlying mechanism, we tested nuclear factor (NF)-kappaB activity and found that NF-kappaB was activated in HCC cells treated with gemcitabine but not in HCC cells that expressed E1A. Occurrence of apoptosis entails cleavage of poly (ADP-ribose) polymerase (PARP), a nuclear protein involved in DNA repair, genome stability, and maintenance of telomere length. Our study showed that gemcitabine enhanced PARP expression. However, E1A did not induce PARP cleavage but rather suppressed PARP expression at the transcriptional level. Further study showed that both NF-kappaB and PARP played protective roles in the prevention of E1A+gemcitabine-induced apoptosis. PMID- 14559809 TI - Conditional activation of fibroblast growth factor receptor (FGFR) 1, but not FGFR2, in prostate cancer cells leads to increased osteopontin induction, extracellular signal-regulated kinase activation, and in vivo proliferation. AB - Changes in the fibroblast growth factor receptor (FGFR) axis are often associated with prostate cancer (CaP) progression. We have used chemically induced dimerization (CID) to elucidate the individual contributions of FGFR1 and FGFR2 to tumor etiology. Novel CaP cell lines stably expressing CID/AP20187-inducible FGFR1 (iFGFR1) and iFGFR2 were made using the tumorigenic transgenic adenocarcinoma of the murine prostate (TRAMP)-derived clone, TRAMP-C2N (C2N), to generate C2N.iFGFR1 or C2N.iFGFR2 cells. To test the effects of iFGFR activation on tumor growth, mice bearing s.c. C2N.iFGFR1- or C2N.iFGFR2-derived tumors were treated biweekly with CID. Activation of iFGFR1 led to rapid tumor growth as a result of increased proliferation. In contrast, expression of iFGFR2 inhibited tumor growth. Furthermore, we have ascertained that FGFR1 activation appears to be most important during the early stages of tumor development, but once established, tumors become rapidly CID independent. In these C2N-based lines, quantitative signaling differences were seen between the two receptors, with iFGFR1 leading to more robust extracellular signal-regulated kinase activation. Additionally, activation of iFGFR1, but not iFGFR2, led to strong up-regulation of osteopontin, a secreted glycoprotein involved in integrin activation and associated with CaP progression and metastasis. These studies support the hypothesis that observed changes in the FGFR axis in mammals during CaP progression are causally important. PMID- 14559810 TI - The prolyl isomerase Pin1 is a novel prognostic marker in human prostate cancer. AB - Prostate cancer (PCa) is the most common male cancer in the United States. A major challenge that remains is to predict the clinical outcome in managing PCa patients. The prolyl isomerase Pin1 has been shown to be overexpressed in some human cancer tissues and thought to be an important player in several oncogenic pathways. However, the relationship between Pin1 expression and clinical outcome of cancer patients has not been explored. In this study, we investigated the role of Pin1 in human PCa progression and its clinicopathological significance. Immunohistochemical assessment with affinity-purified polyclonal Pin1-specific antibodies was performed on formalin-fixed paraffin sections of tissue microarray composed of 580 radical prostatectomy specimens. As determined by visual semiquantitation and confirmed by automated image analysis quantitation, Pin1 expression was positively correlated with clinical stage. Furthermore, Cox survival analysis results indicated that patients with a higher Pin1 expression had a significantly higher probability of recurrence than their counterparts with low Pin1 expression, as defined by a serum prostate-specific antigen level of > or =0.4 ng/ml on two consecutive occasions after radical prostatectomy. In addition, patients with high Pin1 expression had almost 4 times the risk of having earlier recurrence than those with low Pin1 expression; patients with a very high level had 8.1 times the risk of an earlier recurrence than a low Pin1 expresser. Pin1 was also an excellent predictor of recurrence in the subset of patients with Gleason score 6 or 7 when analyzed separately: a patient with high Pin1 expression had 8.6 times the risk of having earlier recurrence than one with low Pin1 expression. Pin1 expression is as good as or better than currently used postoperatively available clinicopathological parameters and potentially could be used in the preoperative setting to assist in choice of treatment. Thus, this study suggests a role for Pin1 expression as a potentially excellent prognostic marker in PCa and suggests that Pin1 may also serve as a novel therapeutic target for PCa. PMID- 14559811 TI - Initial experience in small animal tumor imaging with a clinical positron emission tomography/computed tomography scanner using 2-[F-18]fluoro-2-deoxy-D glucose. AB - The feasibility of small animal imaging using a clinical positron emission tomography/computed tomography (PET/CT) scanner with [F-18]-fluoro-2-deoxy-D glucose (FDG) was evaluated. As tumor-bearing small animal models, rabbits with VX-2 liver tumors, rats with mammary tumors on the back, and mice with LS174T human colon tumor xenografts were prepared. Two-dimensional PET, CT, and fused PET/CT images were obtained and reconstructed with a combined PET/CT system using a conventional protocol for humans and dedicated high-resolution mode protocols specialized for each species. Estimated radioactivity concentrations in tumors and normal organs determined noninvasively on FDG-PET/CT were compared with the actual tissue radioactivity levels determined from gamma-counting after vivisection in rats. In addition, recovery-corrected radioactivity concentrations were calculated and evaluated using the tumor/normal organ sizes measured on CT. Tumors in rabbits and rats were clearly visualized by FDG-PET/CT in the dedicated protocols, and images were considered suitable for research purposes. With the aid of thin-slice CT-mapping images, FDG uptake was correctly localized in the viable tumor regions. In mice, increased FDG uptake in tumors with varying activity levels was observed, but detailed anatomical information was not optimally provided from the images, even using specialized protocols. The estimated radioactivity concentrations of tumors and normal organs were close to the actual radioactivity concentrations obtained by gamma-counting (r = 0.97, P < 0.001, the estimated/actual slope: 1) when recovery correction was applied using the sample sizes measured on CT. FDG-PET/CT imaging with a modern clinical scanner was demonstrated to be feasible, of excellent quality, and quite quantitatively accurate for research in rabbits or rats with tumors of appropriate size (>2 cm without recovery correction and >1 cm with recovery correction). Evaluation of FDG uptake within a tumor was possible with the aid of CT images. Dedicated small animal PET/CT scanner would be better suited for evaluating tumor-bearing mice and likely could enhance imaging smaller tumors in rabbits or rats. Although it has limitations, small animal imaging with a clinical PET/CT scanner may be quite adequate for sequential noninvasive imaging in oncology research because the CT is of high resolution, allowing for localization of PET findings and for more precise noninvasive estimation of radioactivity concentration through partial volume corrections. PMID- 14559812 TI - Glycodelin in ovarian serous carcinoma: association with differentiation and survival. AB - Ovarian cancer consists of many subtypes, serous carcinoma being the most common of them. In addition to the histopathological subtype, grading, clinical staging, and the amount of residual tumor, a great number of putative prognostic markers have been introduced. This study addresses in ovarian serous carcinoma the role of glycodelin, the major progesterone-regulated lipocalin protein of the reproductive axis with diverse actions in cell recognition and differentiation. Glycodelin expression was determined by immunohistochemistry of tissue microarrays in ovarian serous carcinomas from 460 patients, and the results were analyzed with respect to progesterone receptor subtype A (PRA) and progesterone receptor subtype B (PRB), clinical parameters, and survival. Glycodelin was localized to the cytoplasm of tumor cells, whereas vascular endothelium in tumor tissue was glycodelin-negative. Glycodelin expression was more frequent in well differentiated (grade I, 79%) than in poorly differentiated carcinomas (grade III, 51%; P < 0.0001), and it was also more frequent in early-stage compared with advanced-stage carcinomas (P = 0.002). Nuclear PRA and PRB were often coexpressed with cytoplasmic glycodelin. Although this was not consistent in all tumors, there was a positive correlation between the presence of glycodelin and PRs in the tumor (P < 0.02), but not between the presence of, or the absence of, glycodelin in tumor and the CA-125 serum concentration. Although in multivariate analysis glycodelin was not an independent variable, the patients with glycodelin expressing tumors showed a higher 5-year overall survival compared with those with glycodelin-negative tumors (55 versus 39%; P < 0.0001; hazard ratio in univariate analysis, 0.57; confidence interval, 0.44-0.74). This difference was notable in patients with grade I tumors and stage III disease. In the latter group, the 10-year survival probability of patients with glycodelin-positive tumors was more than twice as high as that in women with glycodelin-negative tumors. This was also found within well-defined clinical categories, e.g., stage III/grade II and stage III/grade III carcinomas, in which patients with glycodelin-positive tumors carried significantly better 10-year overall survival compared with those with glycodelin-negative tumors. It is concluded that, in ovarian serous carcinoma, glycodelin expression portends better prognosis, probably because of its differentiation-related disposition. PMID- 14559813 TI - Claudin-4 expression decreases invasiveness and metastatic potential of pancreatic cancer. AB - Claudin-4 has been identified as an integral constituent of tight junctions and has been found to be highly expressed in pancreatic cancer. The aim of the present study was to elucidate the effect of claudin-4 on growth and metastatic potential in pancreatic cancer cells, as well as the regulation of claudin-4 by oncogenic pathways. Claudin-4 was stably overexpressed in SUIT-2 pancreatic cancer cells, and its effect on invasion and growth in vitro was examined by using two-chamber invasion assays, soft agar assays, and fluorescence-activated cell sorter analysis. Claudin-4 localization was characterized by light and electron microscopy, and pulmonary colonization was analyzed in vivo after injection of claudin-4 overexpressing cells into the tail vein of nude mice. Overexpression of claudin-4 was associated with significantly reduced invasive potential in vitro and inhibited colony formation in soft agar assays. In vivo, tail vein-injected claudin-4 overexpressing cells formed significantly less pulmonary metastases in comparison with mock-transfected cells. These effects were not caused by changes in proliferation, cell cycle progression, or matrix metalloproteinase gelatinolytic activity, but were paralleled by increased cell contact formation. Moreover, proinvasive transforming growth factor beta was able to down-regulate claudin-4 in PANC-1 cells. Inhibition of Ras signaling by using dominant-negative Ras and specific inhibitors of both downstream effectors mitogen-activated protein/extracellular signal-regulated kinase kinase and phosphatidylinositol 3'-kinase also decreased claudin-4 expression. Our findings identify claudin-4 as a potent inhibitor of the invasiveness and metastatic phenotype of pancreatic cancer cells, and as a target of the transforming growth factor beta and Ras/Raf/extracellular signal-regulated kinase pathways. PMID- 14559814 TI - c-MET mutational analysis in small cell lung cancer: novel juxtamembrane domain mutations regulating cytoskeletal functions. AB - Small cell lung cancer (SCLC) is an aggressive cancer, and most patients present with cancer already spread beyond the lung. The receptor tyrosine kinase (RTK) c MET has been implicated in various solid tumors, including SCLC, and is involved in mediating tumorigenesis, cell motility, scattering, invasion and metastasis. Mutations of c-Met have been described in renal papillary carcinoma and gastrointestinal cancers including hepatocellular carcinoma. The sequence of c MET was examined for possible mutations in the 10 SCLC cell lines and 32 paired SCLC/normal tissues. Novel c-MET alterations were identified among 3 of 10 separate SCLC cell lines and in 4 of 32 SCLC tumor tissue samples. These include two different c-MET missense mutations in the juxtamembrane (JM) domain (R988C found in NCI-H69 and H249 cell lines; and T1010I in SCLC tumor sample T31). Also, there are one Sema domain missense mutation (E168D in SCLC tumor sample T5), two base-pair insertional mutations (IVS13- (52-53)insCT in both SCLC tumor samples T26 and T27) within the pre-JM intron 13, as well as an alternative transcript involving exon 10 (H128 cell line). c-MET receptors are expressed at various levels among the 10 SCLC cell lines studied (high expression: H69, H345, H510, and H526; medium-expression: H128 and H146; and low/no-expression: H82, H209, H249, and H446). The level of c-MET expression does not have any apparent correlation with presence or absence of mutations of c-MET in the cell lines. We show that the two identified JM mutations (R988C and T1010I), when introduced into the interleukin-3 (IL-3)-dependent BaF3 cell line, regulated cell proliferation resulting in a small but significant growth factor independence. When introduced into a SCLC cell line (H446, with minimal endogenous wild-type c MET expression), the JM mutations also regulated cell morphology and adhesion, as well as causing enhanced tumorigenicity by both increases in focus-formation and soft-agar colony-formation assays. Both of the JM mutations also increased cell motility and migration evident in wound healing assay and time-lapse video microscopy speed analysis. The JM mutations also altered the c-MET RTK signaling, resulting in preferentially increased constitutive tyrosine phosphorylation of various cellular proteins, including the key focal adhesion protein paxillin on tyrosine residue Y31 (first CRKL-binding site), correlating with increased motility. These results suggest a novel and unique role of the JM domain in c-MET signaling in SCLC with significant implications in cytoskeletal functions and metastatic potential. The novel JM gain-of-function somatic mutations described are the first to be reported in SCLC, and may be associated with a more aggressive phenotype. It would now be useful to study the inhibition of c-MET as a therapeutic target against SCLC. PMID- 14559815 TI - Relationship between elevated FX expression and increased production of GDP-L fucose, a common donor substrate for fucosylation in human hepatocellular carcinoma and hepatoma cell lines. AB - The levels of fucosylated glycoproteins in various cancers and inflammatory processes have been a subject of intense study. The level of fucosyltransferases and intracellular GDP-L-fucose, a sugar nucleotide and a common donor substrate for all fucosyltransferases, may regulate the level of fucosylated glycoproteins. This study reports on the determination of GDP-L-fucose levels in human hepatocellular carcinoma (HCC) and surrounding tissues, using a recently established high-throughput assay system. Levels of GDP-L-fucose in HCC tissues were significantly increased compared with adjacent nontumor tissues or normal livers. The mean +/- SD for GDP-L-fucose level was 3.6 +/- 0.2 micro mol/mg in control liver, 4.6 +/- 0.9 micro mol/mg in adjacent noninvolved liver tissues (chronic hepatitis, 4.4 +/- 0.7 micro mol/mg; liver cirrhosis, 4.8 +/- 0.9 micro mol/mg), and 7.1 +/- 2.5 micro mol/mg in HCC tissues. The level of GDP-L-fucose in HCC decreased in proportion with tumor size (r = -0.675, P = 0.0002). When expression of the series of genes responsible for GDP-L-fucose synthesis was investigated, the gene expression of FX was found to be increased in 70% (7 of 10) of the HCC tissues examined compared with that in their surrounding tissues. The levels of GDP-L-fucose were positively correlated with the expression of FX mRNA (r = 0.599, P = 0.0074). The levels of FX gene expression in some human hepatoma and hepatocyte cell lines were determined. FX mRNA production was strongly increased in HepG2 and Chang liver, moderately increased in Hep3B and HLF, and, in HLE, was similar to that of a normal human liver tissue. To investigate the effect of GDP-L-fucose on core fucosylation, FX cDNA was transfected into Hep3B cells, which express a relatively low level of GDP-L fucose:N-acetyl-beta-D-glucosaminide alpha1-6 fucosyltransferase (alpha1-6 FucT) and FX mRNA. Transfection of this gene caused an increase in GDP-L-fucose levels as well as the extent of fucosylation on glycoproteins, including alpha fetoprotein, as judged by reactivity to lectins. Collectively, the results herein suggest that the high level of fucosylation in HCC is dependent on a high expression of FX followed by increases in GDP-L-fucose, as well as an enhancement in alpha1-6 FucT expression. Thus, an elevation in GDP-L-fucose levels and the up regulation of FX expression represent potential markers for HCC. PMID- 14559816 TI - The PCPH oncoprotein antagonizes the proapoptotic role of the mammalian target of rapamycin in the response of normal fibroblasts to ionizing radiation. AB - Exposure of normal mouse fibroblasts (MEF3T3) to ionizing radiation (IR) resulted in a dose-dependent increase of mTOR mRNA and protein levels and the shuttling of the mTOR protein from its normal, predominantly mitochondrial location to the cell nucleus. The same IR doses that activated mTOR induced the phosphorylation of p53 on Ser(18) (mouse equivalent to human Ser(15)) and the subsequent transcriptional activation of PUMA, a known proapoptotic p53-target gene, and promoted apoptosis involving increased overall caspase activity, caspase-3 activation, cleavage of poly(ADP-ribose) polymerase (PARP) and classic protein kinase C (PKC) isoforms, and DNA fragmentation. The proapoptotic role of mTOR in this process was demonstrated by the fact that rapamycin, a mTOR inhibitor, blocked p53 Ser(18) phosphorylation, the induction of PUMA, and all other apoptosis events. Furthermore, the proapoptotic function of mTOR was also antagonized by the expression in MEF3T3 cells of the PCPH oncoprotein, known to enhance cell survival by causing partial ATP depletion. Tetracyclin (Tet) regulated expression of oncogenic PCPH, or overexpression of normal PCPH, blocked both phosphorylation and nuclear shuttling of mTOR in response to IR. These results indicate that alterations in PCPH expression may render tumor cells resistant to IR, and perhaps other DNA-damaging agents, by preventing mTOR activation and signaling. PMID- 14559817 TI - Methylation-associated silencing of the thrombospondin-1 gene in human neuroblastoma. AB - Tumor angiogenesis, a major requirement for tumor outgrowth and metastasis, is regulated by pro- and antiangiogenic factors. Methylation-associated inactivation of the angiogenesis inhibitor thrombospondin-1 (TSP-1) has been observed recently in some adult tumors. To investigate the role of TSP-1 in pediatric cancer, we examined its pattern of expression and mechanisms of regulation in neuroblastoma (NB). TSP-1 was silenced in a subset of undifferentiated, advanced-stage tumors and NB cell lines. In contrast, most localized tumors expressed this angiogenesis inhibitor, and a significant correlation between morphological evidence of neuroblast differentiation and TSP-1 expression was observed. Luciferase assays demonstrated the presence of nuclear factors required for TSP-1 transcription in both TSP-1-positive and -negative cell lines, but no correlation between TSP-1 promoter activity and the level of TSP-1 mRNA expression was seen. Our studies indicate that the transcriptional silencing of TSP-1 was caused by methylation. TSP-1 promoter methylation was detected in all of the NB cell lines lacking TSP-1 mRNA and in 37% of the NB clinical tumors analyzed. Furthermore, treatment with the demethylating agent, 5-Aza-2'-deoxycytidine (5-Aza-dC), restored TSP-1 expression in NB cell lines. Disrupting methylation with 5-Aza-dC also led to significant inhibition of NB in vivo and re-expression of TSP-1 in a subset of NB xenografts. These results suggest that 5-Aza-dC inhibits NB growth by augmenting the expression of TSP-1 along with other genes that suppress tumor growth. Demethylating agents may prove to be effective candidates for the treatment of children with NB. PMID- 14559818 TI - The intrinsic mitochondrial membrane potential (Deltapsim) is associated with steady-state mitochondrial activity and the extent to which colonic epithelial cells undergo butyrate-mediated growth arrest and apoptosis. AB - Transformation of colonic epithelial cells is characterized by decreased mitochondrial activity, increased mitochondrial membrane potential (Deltapsi(m)), and disruptions in the equilibrium between cell proliferation and death by apoptosis. We have previously shown that an intact Deltapsi(m) is essential for growth arrest and apoptosis induced by butyrate, a physiological regulator of maturation in these cells, suggesting a role for the Deltapsi(m) in the initiation and integration of proliferation and apoptotic pathways. To extend this work, we have generated isogenic cell lines, from SW620 human colonic carcinoma cells, which exhibit significant differences in intrinsic Deltapsi(m). These differences in Deltapsi(m) are not linked to alterations in viability, Bcl 2 levels, or the differentiation status of the cells. However, compared with parental cells and those with increased Deltapsi(m), cells with decreased intrinsic Deltapsi(m) exhibit significantly higher levels of steady-state mitochondrial mRNA and butyrate-induced p21(WAF1/Cip1) and G(0)-G(1) arrest. Moreover, despite butyrate-mediated translocation of proapoptotic Bax and Bak to the mitochondria, fewer cells with elevated intrinsic Deltapsi(m) exhibit concomitant cytochrome c release, and cells with elevated Deltapsi(m) undergo significantly lower levels of Deltapsi(m) dissipation and apoptosis than parental cells, or cells with decreased Deltapsi(m). Homeostasis of the colonic mucosa depends on balancing cell proliferation with apoptosis, and mitochondrial abnormalities are associated with disruptions in this balance. Thus, by affecting steady-state mitochondrial activity and the extent to which cells enter growth arrest and apoptotic cascades, these data establish a role for the intrinsic Deltapsi(m) in contributing to the probability of colonic tumorigenesis and progression. PMID- 14559819 TI - Involvement of TSLC1 in progression of esophageal squamous cell carcinoma. AB - Frequent allelic losses of 11q23 in esophageal squamous cell carcinoma (ESCC) have been reported previously, but no tumor suppressor genes in this region have been identified in ESCC. TSLC1 was identified on chromosome 11q23.2 as a tumor suppressor gene in non-small cell lung cancer by functional complementation of a lung adenocarcinoma cell line. The purpose of this study is to evaluate the role of TSLC1 in ESCC. Loss of TSLC1 expression was observed by reverse transcription PCR in 75% of the cell lines (27 of 36) and 50% of the primary tumors from ESCC patients (28 of 56). In a clinicopathological analysis, loss of TSLC1 expression correlated significantly with depth of invasion (pT) and status of metastasis (pM; P = 0.012 and 0.036, respectively). Patients with tumors lacking TSLC1 expression tended to have a poorer prognosis than those with tumors expressing TSLC1. (P = 0.079). Moreover, TSLC1 expression was an independent prognostic factor in a multivariate analysis (P = 0.049). Methylation analyses revealed that TSLC1 expression or loss correlated with the promoter methylation status, as determined by bisulfite sequencing, and that TSLC1 expression could be restored by a demethylating agent in certain cell lines. The growth of TSLC1-transfected ESCC cells was significantly suppressed both in vitro and in vivo (P < 0.01), possibly by a G(1) cell cycle arrest. TSLC1 expression also suppressed motility and invasion of ESCC cells in vitro significantly (P < 0.01). These findings suggest that loss of TSLC1 expression has an important role in tumor growth, cell motility, and invasion and is associated with aggressive tumor behavior in ESCC. PMID- 14559820 TI - Radiofrequency thermal ablation sharply increases intratumoral liposomal doxorubicin accumulation and tumor coagulation. AB - Combining radiofrequency (RF) ablation with i.v. liposomal doxorubicin (Doxil) increases intratumoral doxorubicin accumulation and tumor destruction. The purpose of this study was to characterize and better define the specific parameters of such treatment in an animal tumor model. Four hundred R3230 mammary adenocarcinoma nodules were implanted in 250 Fischer rats. First, paired tumors received combined standardized RF (70 degrees C +/- 2 degrees C, 5 min) followed 30 min later with i.v. Doxil (1 mg) or Doxil alone. Intratumoral doxorubicin uptake was evaluated using fluorospectrophotometry 2-120 h after therapy (n = 110). The effects of varying i.v. Doxil doses (0.0625-7.0 mg; n = 100) and the RF tip temperatures (45 degrees C-90 degrees C; n = 190) on subsequent intratumoral doxorubicin uptake and induced tumor necrosis were evaluated. Intratumoral doxorubicin accumulation increased to a maximum at 72 h with greater uptake in the RF-ablated tumors compared with controls (P < 0.01). Greater dose-dependent intratumoral doxorubicin increases (to 37.3 +/- 7.7 microg/g) were seen with combined RF/Doxil therapy (P < 0.01). RF ablation reduced the i.v. Doxil dose needed to achieve intratumoral doxorubicin uptake of 13 microg/g from 7 to 2 mg. Increasing tip temperatures from 50 degrees C to 90 degrees C increased the ratio of doxorubicin in RF to nonablated tumors from 1.2 +/- 0.4 to 5.9 +/- 3.8 (P < 0.01). At all temperatures, greater tumor necrosis was identified for RF/Doxil treated tumors compared with tumors treated with RF alone (P < 0.05). The threshold for inducing necrosis was 5 degrees C lower for tumors receiving combined therapy (P < 0.01). RF tumor ablation sharply increases intratumoral Doxil accumulation over i.v. Doxil alone, enabling a reduction of systemic dose while obtaining higher intratumoral concentrations than otherwise achievable. Combined therapy also increases tumor destruction over either therapy alone. PMID- 14559821 TI - SH2 domain containing protein tyrosine phosphatase 2 regulates concanavalin A dependent secretion and activation of matrix metalloproteinase 2 via the extracellular signal-regulated kinase and p38 pathways. AB - We investigated the role of SH2 domain containing protein tyrosine phosphatase (SHP) 2 in Concanavalin A (Con A) -dependent signaling that leads to the augmented secretion and activation of matrix metalloproteinase (MMP) 2. In cells expressing mutant SHP-2 in which 65 amino acids in the SH2-N domain were deleted, we found that production, secretion, and proteolytic activation of MMP-2 in response to Con A treatment was severely impaired. Under Con A stimulation, complex formation of SHP-2 with SOS-1 and Grb-2 together with the activation of Ras signaling was clearly observed in wild-type cells, but not in SHP-2 mutant cells. In wild-type cells, Con A-treatment activated dual signaling pathways, extracellular signal-regulated kinase (Erk) and p38, in a Ras-dependent manner, whereas Con A-dependent activation of these signaling pathways was absent in SHP 2 mutant cells. In addition, pretreatment of wild-type cells with U0126, a potent inhibitor for mitogen-activated protein/ERK kinase 1, or with SB203580, a specific inhibitor for p38, significantly inhibited the Con A-dependent secretion and activation of MMP-2. However, overexpression of active mitogen-activated protein/ERK kinase 1 in SHP-2 mutant cells could not induce clear activation of MMP-2 secretion, although these cells responded well to the Con A treatment in a p38-dependent manner. Finally, reintroduction of wild-type SHP-2 into SHP-2 mutant cells rescued Erk and p38 activation, and also MMP-2 secretion, whereas dominant-negative SHP-2 could block the Con A-dependent activation of Erk and p38. Taken together, our results strongly suggest that SHP-2 plays a critical role as a positive mediator for Con A-dependent activation of MMP-2 secretion via Ras-Erk and Ras-p38 signalings. PMID- 14559822 TI - Caspase-mediated cleavage converts Livin from an antiapoptotic to a proapoptotic factor: implications for drug-resistant melanoma. AB - Inhibitor of apoptosis protein (IAP) is a family of intracellular proteins that plays an essential role in the regulation of apoptosis. Recently, we and others discovered a new member of this family, termed Livin. Many studies have focused on the inhibitory effect of IAPs on caspases. Here, we describe a novel regulatory mechanism by which Livin is cleaved by the caspases. Strikingly, the cleaved Livin, although containing intact baculovirus IAP repeat and RING domains, does not only lose its antiapoptotic function but also gains a proapoptotic effect. The cleavage is site specific at Asp-52 and is restricted to effector caspase-3 and -7. Most importantly, we demonstrate the role of Livin and this regulatory mechanism in the drug resistance of melanoma patients. Using primary cultures derived from melanoma patients, we found a correlation between Livin overexpression, in vitro drug resistance, and the patient's clinical response. PMID- 14559823 TI - Coexpression of Helicobacter pylori's proteins CagA and HspB induces cell proliferation in AGS gastric epithelial cells, independently from the bacterial infection. AB - Adenocarcinoma of the stomach is the second most common cause of cancer mortality in the world. The purpose of this study was to evaluate the potential role in carcinogenesis of two secreted Helicobacter pylori's proteins, CagA and HspB, both shown to increase the risk of gastric carcinoma in patients infected with H. pylori-positive strain. The effects of these two proteins on cell kinetics and the ability to selectively affect the expression of cell cycle-related proteins by transfection of a human gastric epithelial cell line (AGS) were analyzed. Using a genomic library of H. pylori, we isolated and cloned CagA and HspB. The effects of the overexpression of these proteins on cell growth were analyzed in AGS cells by immunoblots, proliferation assay, and flow cytometry. Coexpression of CagA and HspB in AGS cells in the first 48 h caused an increase of the level of E2F transcription factor, cyclin D3, and phosphorylated retinoblastoma protein, all involved in the G(1)-S checkpoint of the cell cycle. Consistently, an increase of cell proliferation, corresponding to an augment of the fraction of the cells in the S-G(2)-M phase of the cell cycle, was also demonstrated. Moreover, an increase of c-jun protein levels, but not of c-fos, was also found after coexpression of CagA and HspB. All these data suggest that CagA and HspB, independently from the bacterial infection, have a direct effect on the cell growth of the gastric cells acting on the G(1)-S checkpoint of the cell cycle. PMID- 14559824 TI - PTEN reverses MDM2-mediated chemotherapy resistance by interacting with p53 in acute lymphoblastic leukemia cells. AB - The tumor suppressor PTEN has been associated with the cellular localization of MDM2 in regulation of apoptosis through inhibiting PI3k/Akt signaling. To investigate whether expression of PTEN is involved in MDM2-mediated chemoresistance, we examined a set of acute lymphoblastic leukemia (ALL) cell lines for the expression of PTEN and sensitivity to doxorubicin. Testing 9 ALL cell lines selected for wild-type p53 phenotype and uniformly high levels of MDM2 expression, we initially demonstrated that cell lines with high levels of PTEN expression were sensitive to doxorubicin, whereas lines lacking PTEN expression were generally resistant. Forced expression of PTEN in a PTEN-negative and doxorubicin-resistant ALL line (EU-1) resulted in decreased cell growth and enhanced sensitivity to doxorubicin. Examining the cellular localization of MDM2, we confirmed that the majority of MDM2 is localized in the nucleus in PTEN negative doxorubicin-sensitive ALL cells, whereas MDM2 is expressed predominantly in the cytoplasm in either PTEN-positive or PTEN-transfected cells. Furthermore, by coimmunoprecipitaton and cotransfection assays, we found that PTEN physically binds p53 in vitro as well as in vivo. Binding of PTEN to p53 attenuated MDM2 mediated p53 inhibition. These results suggest that PTEN inhibits MDM2 and protects p53 through both p13k/Akt-dependent and -independent pathways. Furthermore, loss of PTEN can result in resistance to apoptosis by activating MDM2-mediated antiapoptotic mechanism. PMID- 14559825 TI - Reversal of tumorigenicity and the block to differentiation in erythroleukemia cells by GATA-1. AB - Oncogenic transformation usually inhibits normal cell differentiation processes. Certain chemical agents can force some tumor cells to resume their differentiation program and undergo cell cycle arrest, an approach termed differentiation therapy. Mouse erythroleukemia (MEL) cells represent an important cell culture model system for investigating the principles of differentiation therapy. MEL cells are malignant erythroblasts that are blocked from differentiating into mature erythroid cells because of inappropriate expression of the transcription factor PU.1, which binds to and represses GATA-1, a key transcriptional stimulator of red blood cell differentiation. We report here that the block to differentiation in MEL cells can be overcome by providing the cells with additional GATA-1. A conditionally active form of GATA-1 can trigger the cells to differentiate, undergo terminal cell division, and lose their tumorigenicity. We also show that the gene for the cell cycle inhibitor p21 is transcriptionally regulated by GATA-1 and is a likely downstream effector of GATA 1 that helps to promote differentiation and proliferation arrest. PMID- 14559826 TI - Decreased mitochondrial nitric oxide synthase activity and hydrogen peroxide relate persistent tumoral proliferation to embryonic behavior. AB - Differential expression and activity of constitutive mitochondrial nitric oxide synthase (mtNOS) in the mitochondrial compartment is followed by significant variations in matrix nitric oxide (NO) steady-state concentration. The mitochondrial utilization of NO involves the production of superoxide anion and H(2)O(2), a species freely diffusible outside the mitochondria that participates in the modulation of cell proliferation and apoptosis and in cell transformation and cancer. On these bases, we analyzed the modulation of mtNOS in the frame of cellular redox state in M3, MM3, and P07 murine tumors and their respective cell lines, as compared with normal proliferating and quiescent tissues. The results showed that: (a) tumoral and proliferating mitochondria only retain 10-50% of the activity of complexes I, II-III, and IV and Mn-SOD of quiescent tissues; (b) normal proliferating tissues, like embryonic liver or pregnant mammary gland, have 10-20% of mtNOS expression and activity and mitochondrial H(2)O(2) yield than quiescent nonproliferating tissues; (c) similarly but irrespective of mtNOS expression, tumoral mitochondria have no >5% of mtNOS activity and H(2)O(2) yield of mature tissues; and (d) in opposition to stable tissues, both tumoral and normal proliferating cells exhibit high cyclin D1 expression and low pro apoptotic p38mitogen-activated protein kinase activity. Dually, H(2)O(2) stimulated tumor cell proliferation (<10 microM) or markedly inhibited it (>10 microM) with parallel variations of cyclin D1, phospho-extracellular-regulated kinase1/2, and phospho-p38mitogen-activated protein kinase. It is surmised that decreased oxidative phosphorylation, defective tumoral mtNOS, and low mitochondrial NO-dependent H(2)O(2) may be a platform to link persistent tumoral growth to embryonic behavior. PMID- 14559827 TI - Intratumoral delivery of dendritic cells engineered to secrete both interleukin (IL)-12 and IL-18 effectively treats local and distant disease in association with broadly reactive Tc1-type immunity. AB - Dendritic cells (DCs) were adenovirally engineered to constitutively and durably secrete the potent Th1-biasing cytokines interleukin (IL)-12 (AdIL12DC) and/or IL 18 (AdIL18DC) and evaluated for their ability to promote therapeutic antitumor immunity in murine sarcoma models. Injection of either AdIL12DC or AdIL18DC into day 7 CMS4 or MethA tumors resulted in tumor rejection or slowed tumor growth when compared with control cohorts. Importantly, intratumoral injection with DCs engineered to secrete both IL-12 and IL-18 (AdIL12/IL18DC) resulted in complete and the most acute rejection of any treatment group analyzed. This strategy was also effective in promoting the regression of contralateral, untreated tumors. Both CD4+ and CD8+ T cells were required for tumor rejection. CD8+ splenic T cells from mice treated with AdIL12/IL18DC produced the highest levels of IFN gamma in response to tumor rechallenge in vitro and displayed the broadest repertoire of Tc1-type reactivity to acid-eluted, tumor-derived peptides among all treatment cohorts. This apparent enhancement in cross-presentation of tumor associated epitopes in vivo may result from the increased capacity of engineered DCs to kill tumor cells, survive tumor-induced apoptosis, and present immunogenic MHC/tumor peptide complexes to T cells after intratumoral injection. In support of this hypothesis, cytokine gene-engineered DCs expressed higher levels of MHC and costimulatory molecules, as well as Fas ligand and membrane-bound tumor necrosis factor alpha, with the latter markers associated with elevated tumoricidal activity in vitro. Cytokine gene-engineered DCs appeared to have a survival advantage in situ when injected into tumor lesions, to be found in approximation with regions of tumor apoptosis, and to have the capacity to ingest apoptotic tumor bodies. These results support the ability of combined cytokine gene transfer to enhance multiple effector functions mediated by intralesionally injected DCs that may concertedly promote cross-priming and the accelerated immune-mediated rejection of tumors. PMID- 14559828 TI - E-selectin up-regulation allows for targeted drug delivery in prostate cancer. AB - We have used the Eos Hu03 GeneChip array, which represents over 92% of the transcribed human genome, to measure gene expression in a panel of normal and diseased human tissues. This analysis revealed that E-selectin mRNA is selectively overexpressed in prostate cancer epithelium, a finding that correlated strongly with E-selectin protein expression as assessed by immunohistochemistry. Antibodies against E-selectin that blocked function failed to impede cancer cell growth, suggesting that overexpression of E-selectin was not essential for cell growth. However, a novel auristatin E-based antibody drug conjugate (ADC), E-selectin antibody valine-citrulline monomethyl-auristatin E, was a potent and selective agent against E-selectin-expressing cancer cell lines in vitro, with the degree of cytotoxicity varying with surface antigen density. Interestingly, sensitivity to the ADC differed among cell lines from different tissues expressing similar amounts of E-selectin and was found to correlate with sensitivity to free auristatin E. Furthermore, E-selectin-expressing tumors grown as xenografts in severe combined immunodeficient mice were responsive to treatment with E-selectin antibody valine-citrulline monomethyl-auristatin E in vivo, with more than 85% inhibition of tumor growth observed in treated mice. These findings demonstrate that an E-selectin-targeting ADC has potential as a prostate cancer therapy and validates a genomics-based paradigm for the identification of cancer-specific antigens suitable for targeted therapy. PMID- 14559829 TI - Inhibition of wild-type and mutant Bcr-Abl by pyrido-pyrimidine-type small molecule kinase inhibitors. AB - Imatinib mesylate (STI571, Glivec), a 2-phenylaminopyrimidine small-molecule ATP competitor-type kinase inhibitor, proved to be active in Philadelphia-positive leukemias. Resistance toward imatinib develops frequently in advanced-stage Philadelphia-positive leukemia, and is even observed in chronic-phase chronic myelogenous leukemia. Point mutations within the BCR-ABL kinase domain emerged as a major mechanism of resistance toward imatinib. Mutations occur at positions that determine specific contacts of imatinib to the ATP-binding site. We aimed to examine whether pyrido-pyrimidine-type kinase inhibitors were capable of inhibiting both wild-type and mutant forms of BCR-ABL. We screened 13 different pyrido-pyrimidine with cells expressing wild-type and mutant BCR-ABL. All of the substances specifically suppressed the Bcr-Abl dependent phenotype and inhibited Bcr-Abl kinase activity with higher potency than imatinib. Two of the most active compounds were PD166326 and SKI DV-M016. Interestingly, these compounds suppressed the activation loop mutant Bcr-Abl H396P as effectively as wild-type Bcr-Abl. In addition, nucleotide-binding loop mutations (Y253H, E255K, and E255V) were selectively and potently inhibited. In contrast, T315I, a mutant located at a position that makes a direct contact with imatinib, was not affected. This observation is consistent with the hypothesis that unlike imatinib, pyrido pyrimidine inhibitors bind Bcr-Abl regardless of the conformation of the activation loop. We conclude that pyrido-pyrimidine-type kinase inhibitors are active against different frequently observed kinase domain mutations of BCR-ABL that cause resistance toward imatinib. Resistance as a consequence of selection of mutant BCR-ABL by imatinib may be overcome using second-generation kinase inhibitors because of their higher potency and their ability to bind Bcr-Abl irrespective of the conformation of the activation loop. PMID- 14559830 TI - Angiogenesis inhibitors target the endothelial cell cytoskeleton through altered regulation of heat shock protein 27 and cofilin. AB - Inhibition of angiogenesis has emerged as a key focus for the treatment of cancer, necessitating a better understanding of the downstream molecular targets of angiogenesis inhibitors. Endostatin, thrombospondin-1, fumagillin, and its synthetic derivative, TNP-470, are potent inhibitors of endothelial cell proliferation and migration in culture and of angiogenesis in vivo. To identify targets that mediate the effects of these inhibitors, we compared two-dimensional gel electrophoresis patterns from lysates of treated and untreated human endothelial cells. Among the proteins identified were cofilin and hsp27, two proteins involved in actin dynamics. Western blotting and immunofluorescence experiments confirmed that the phosphorylation states and subcellular localization of these two proteins were affected by all of the inhibitors tested and that treated cells had a more extensive network of actin stress fibers and more numerous focal adhesion plaques compared with untreated cells. Endothelial monocyte activating polypeptide II, another angiogenesis inhibitor, elicited the same response in the actin cytoskeleton and focal adhesions of endothelial cells. This more adherent phenotype may explain the shared ability of these inhibitors to block endothelial migratory signals. Starting with a proteomics approach, we have identified common effector molecules used by a panel of angiogenesis inhibitors that perturb the cytoskeleton to prevent endothelial migration. PMID- 14559831 TI - An adenosine analogue, IB-MECA, down-regulates estrogen receptor alpha and suppresses human breast cancer cell proliferation. AB - Adenosine, a natural metabolite, plays important roles in several physiological and pathological processes, including modulation of cellular proliferation. Here, we report that among different adenosine analogues tested, micromolar concentrations of the A(3) adenosine receptor (A(3)AR)-selective agonist N(6)-(3 iodobenzyl)adenosine-5'-N-methyluronamide (IB-MECA) completely inhibited the growth of the human breast cancer cell lines MCF-7 and ZR-75 while inducing apoptosis in T47D and Hs578T cells, which do not express A(3)AR mRNA. In MCF-7 cells, A(3)AR overexpression did not increase the sensitivity to drug treatment and an A(3)AR antagonist did not abolish IB-MECA effect. In search for mechanisms of the effect of this ligand, we found that in estrogen receptor alpha (ERalpha) positive cells, IB-MECA rapidly down-regulated ERalpha at mRNA and protein levels and consequently at the transcriptional activity level. Moreover, overexpression of ERalpha in MCF-7 cells alleviated the proliferation inhibition induced by IB MECA. The inhibitory effects on cell growth and to some extent on ERalpha were mimicked by 2-chloro-adenosine >3'-deoxyadenosine> adenosine but not by a variety of other ligands. Our studies indicate that IB-MECA can down-regulate ERalpha and inhibit proliferation or induce apoptosis in different breast cancer cell types and raise the possibility of using this and related compounds in breast cancer treatment. PMID- 14559833 TI - Genetic blockade of the insulin-like growth factor-I receptor: a promising strategy for human pancreatic cancer. AB - Pancreatic cancer is one of the most lethal malignant tumors. Insulin-like growth factor (IGF)-I receptor (IGF-Ir) signaling is required for maintenance of growth and tumorigenicity of many tumors, but this pathway has not been well studied in pancreatic cancer. We have shown previously successful therapy in colorectal and lung cancer xenograft models using recombinant adenoviruses expressing dominant negative IGF-I receptors. In this study, we sought to better dissect the mechanism of action of this virus and determine whether IGF-Ir targeted adenoviruses represent potentially effective therapeutics for human pancreatic cancer cells. Truncated IGF-I receptors (IGF-Ir/dn; 482 and 950 amino acids long, respectively, IGF-Ir/482st and IGF-Ir/950st) that function as dominant negative inhibitor were cloned into recombinant adenoviruses and used to treat human pancreatic cancer cells. We assessed the effect of IGF-Ir/dn on signaling blockade, growth, stress response, chemotherapy, radiation-induced apoptosis, and in vivo therapeutic efficacy in xenografts. IGF-Ir/dn expression suppressed tumorigenicity both in vitro and in vivo and up-regulated stressor-induced apoptosis. It effectively blocked both IGF-I and IGF-II-induced activation of Akt 1. IGF-Ir/dn expression increased radiation and chemotherapy-induced apoptosis, and the combination therapy of IGF-Ir/dn with chemotherapy was very effective against tumors in mice. In an i.p. model, IGF-Ir/dn therapy reduced dissemination and prolonged survival times. Moreover, IGF-Ir/482st was more effective than IGF Ir/950st because of its bystander effect. The antitumor activity of IGF-Ir/dn is mediated through inhibition of Akt-1 and enhances the efficacy of chemotherapy. Adenovirus-IGF-Ir/482st may be a useful anticancer therapeutic for pancreatic cancer. PMID- 14559832 TI - Interferon regulatory factor 5, a novel mediator of cell cycle arrest and cell death. AB - We have previously shown a critical role for IFN regulatory factor 5 (IRF-5) in the innate immune response to virus infection. For the first time, we now show that although IRF-5 is a direct target of p53, its cell cycle regulatory and proapoptotic effects are p53 independent. IRF-5 inhibits both in vitro and in vivo B-cell lymphoma tumor growth in the absence of wild-type p53. The molecular mechanism(s) of IRF-5-mediated growth inhibition is associated with a G(2)-M cell cycle arrest and modulation of growth regulatory and proapoptotic genes, including p21, Bak, DAP kinase 2, and Bax. Taken together, these data indicate that although IRF-5 is a downstream target of p53, its growth inhibitory and proapoptotic effects are independent of p53. PMID- 14559834 TI - Inhibition of estrone sulfate-induced uterine growth by potent nonestrogenic steroidal inhibitors of steroid sulfatase. AB - The present study describes the biological in vitro and in vivo evaluation of 2 methoxy derivatives of estrogenic inhibitors of steroid sulfatase, namely 3 sulfamoyloxy-17alpha-p-tert-butylbenzyl(or benzyl)-1,3,5 (10)-estratrien-17beta ols. The addition of the 2-methoxy group conserves the potent inhibitory effect on steroid sulfatase activity (IC(50)s of 0.024 and 0.040 nM) while removing the estrogenic action. Using an ovariectomized mouse model, we show that the first generation of steroid sulfatase inhibitors tested, 3-sulfamoyloxy-17alpha-p-tert butylbenzyl(or benzyl)estra-1,3,5 (10)-trien-17beta-ols and estrone-3-O sulfamate, are estrogenic compounds stimulating estrogen-sensitive uterine growth. Interestingly, the 2-methoxy-3-sulfamoyloxy-17alpha-benzylestra-1,3,5 (10)-trien-17beta-ol (7) has no estrogenic activity but efficiently blocks (s.c. and p.o.) uterine growth induced by estrone sulfate, which is converted into estrone and then estradiol by steroid sulfatase and type 1 17beta-hydroxysteroid dehydrogenase, respectively. This report clearly shows that a steroid sulfatase inhibitor can efficiently block estrogen action from the inactive precursor estrone sulfate, in vitro and in vivo. PMID- 14559835 TI - The breast cancer resistance protein (Bcrp1/Abcg2) restricts exposure to the dietary carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine. AB - The food carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is the most abundant heterocyclic amine found in various protein containing foods. PhIP is mutagenic and carcinogenic in rodents, inducing lymphomas in mice and colon, mammary and prostate carcinomas in rats. It has also been implicated in human breast carcinogenesis. Humans on a normal Western diet are exposed to PhIP on a daily basis. The breast cancer resistance protein (BCRP/ABCG2) transports various anticancer drugs from cells, causing multidrug resistance. By its presence in the apical membrane of the intestine and bile canalicular membrane, it also protects the body from substrate drugs and toxins. We investigated whether Bcrp1 could affect PhIP exposure of the body because this could implicate BCRP activity in the cancer risk due to PhIP. Using polarized cell lines, we found that PhIP is efficiently transported by murine Bcrp1. In vivo pharmacokinetic studies showed that at a dose of 1 mg/kg [(14)C]PhIP the area under the curve for oral administration was 2.9-fold higher in Bcrp1(-/-) compared with wild-type mice (306 +/- 39 versus 107 +/- 15 h.ng/ml) and, for i.v. administration, 2.2 fold higher (386 +/- 36 versus 178 +/- 8.9 h.ng/ml). Wild-type mice cleared [(14)C]PhIP mainly by fecal excretion, but this shifted to primarily urinary excretion in Bcrp1(-/-) mice. In mice with a cannulated gall bladder, both hepatobiliary and direct intestinal excretion of [(14)C]PhIP were greatly impaired in Bcrp1(-/-) compared with wild-type mice (9.0 +/- 4.4 versus 36.5 +/- 9.4% and 1.5 +/- 0.8 versus 4.2 +/- 1.5%, respectively). We conclude that Bcrp1 effectively restricts the exposure of mice to ingested PhIP by decreasing its uptake from the gut lumen and by mediating hepatobiliary and intestinal elimination. Intra- or interindividual differences in BCRP activity in humans may thus also affect the exposure to PhIP and related food carcinogens, with possible implications for cancer susceptibility. PMID- 14559836 TI - Effective therapy for a murine model of adult T-cell leukemia with the humanized anti-CD52 monoclonal antibody, Campath-1H. AB - Adult T-cell leukemia (ATL) develops in a small proportion of human T-cell leukemia virus I-infected individuals. Presently, there is no effective therapy for ATL. A murine model of ATL was produced by introducing leukemic cells (MET-1) from an ATL patient into nonobese diabetic/severe combined immunodeficient mice. The MET-1 cells are activated T cells that express CD2, CD3, CD4, CD25, CD122, and CD52. We evaluated the efficacy of Campath-1H (alemtuzumab; a humanized monoclonal antibody directed to CD52), alone and in combination with humanized anti-Tac (HAT) directed to CD25 (interleukin 2 receptor alpha) or with MEDI-507 directed to CD2. We observed that four weekly treatments with 4 mg/kg HAT significantly prolonged survival of MET-1-bearing mice. However, the survival of mice receiving 4 weeks of 4 mg/kg Campath-1H was significantly longer than that of the group receiving four weekly treatments with HAT (P < 0.001). Treatment with Campath-1H for 4 weeks led to a striking prolongation of the survival of MET 1 ATL-bearing mice that was comparable with that of tumor-free nontreated controls. Using Fc receptor (FcR) gamma(-/-) mice, we found that FcRgammas on polymorphonuclear leukocytes and monocytes are required for Campath-1H-mediated tumor killing in vivo. These results demonstrate that Campath-1H has therapeutic efficacy on ATL in vivo in that the life span of the Campath-1H treatment group was comparable with that of mice that did not receive a tumor or therapy. The main tumor killing mechanism with Campath-1H in vivo involves FcRgamma-containing receptors (e.g., FcRgammaIII) on polymorphonuclear leukocytes and macrophages that mediate antibody-dependent cellular cytotoxicity and/or trigger cross linking induced apoptosis. This study provides support for a clinical trial of Campath-1H in the treatment of patients with T-cell leukemias and lymphomas. PMID- 14559837 TI - Inhibitory effect of the salmosin gene transferred by cationic liposomes on the progression of B16BL6 tumors. AB - Salmosin is a novel disintegrin containing the Arg-Gly-Asp sequence that significantly inhibits platelet aggregation, basic fibroblast growth factor induced endothelial cell proliferation, and tumor progression by antagonizing integrin-mediated cell interactions. Previously, it was shown that daily administration of salmosin was able to inhibit tumor-derived angiogenesis and adherence and proliferation of tumor cells, resulting in suppression of tumor progression. However, it is very difficult to maintain a therapeutic level of salmosin in the blood by systemic administration of the protein. Hence, an alternative strategy for antiangiogenic cancer therapy, based on the in vivo expression of the salmosin gene administered with cationic liposomes, was investigated. The salmosin peptides expressed in vitro inhibited the proliferation of bovine capillary endothelial cells in a dose-dependent manner, presumably as a result of inhibition of cell adhesion mediated via alpha(v)beta(3) integrin. Subcutaneous administration of the salmosin gene resulted in systemic expression of the gene product and concomitant inhibition of the growth of B16BL6 melanoma cells. Suppression of pulmonary metastases, verified by experimental and spontaneous metastasis models in mice, also resulted from salmosin gene treatment. These results suggest that administration of the salmosin gene complexed to cationic liposomes is effective in maintaining antiangiogenic salmosin at an effective therapeutic level and may be clinically applicable to anticancer gene therapy. PMID- 14559838 TI - Neutrophils contribute to the measles virus-induced antitumor effect: enhancement by granulocyte macrophage colony-stimulating factor expression. AB - To investigate the contribution of a neutrophil response to the oncolytic effect of replicating attenuated measles virus (MV), MVs expressing murine granulocyte macrophage colony-stimulating factor (GM-CSF) were generated. The growth characteristics and kinetics of GM-CSF production of these viruses were characterized in vitro. Their biological effects were characterized in mice transgenic for the MV receptor CD46. The oncolytic efficacy of MV GM-CSF was then compared with that of a parental MV and a control, UV-irradiated MV using a human lymphoid tumor model in immunodeficient mice. Intratumoral injection of MV resulted in significant tumor regression or slowing of progression compared with injection of the control. Injection of MV GM-CSF further enhanced the oncolytic effect. In additional experiments, the cellular response to MV, MV GM-CSF, recombinant murine GM-CSF alone, or untreated tumors was quantified. The predominant response was an influx of neutrophils. Intratumoral natural killer cells and macrophages were not detected. The magnitude of the neutrophil response correlated well with tumor regression. Our studies suggest that therapy with replicating MV stimulates a strong neutrophil antitumor response, which can be cytokine-enhanced to improve oncolysis. PMID- 14559839 TI - Selectively increased expression and functions of chemokine receptor CCR9 on CD4+ T cells from patients with T-cell lineage acute lymphocytic leukemia. AB - In a total of 38 typical T-cell lineage acute lymphocytic leukemia (T-ALL) and T cell lineage chronic lymphocytic leukemia (T-CLL) cases investigated, we found that CC chemokine receptor CCR9 was selectively and frequently expressed on T-ALL CD4+ T cells, was moderately expressed on T-CLL CD4+ T cells, and was rarely expressed on normal CD4+ T cells. These findings were demonstrated at protein and mRNA levels using flow cytometry and real-time quantitative reverse transcription PCR technique and were verified by digital confocal microscopy and Northern blotting. Thymus-expressed chemokine, a ligand for CCR9, selectively induced T ALL CD4+ T-cell chemotaxis and adhesion. Interleukin (IL)-2 and IL-4, together, down-regulated the expression and functions of CCR9 in T-ALL CD4+ T cells including chemotaxis and adhesion. It was also demonstrated that IL-2 and IL-4, together, internalized CCR9 on T-ALL CD4+ T cells and subsequently inhibited functions of CCR9 in these cells. Thymus-expressed chemokine mRNA was highly expressed in CD4+ T cells, involving lymph node and skin in T-ALL patients, and was expressed at moderate levels in lymph node and skin tissues in T-CLL patients. Our findings may provide new clues to understanding various aspects of T-ALL CD4+ T cells, such as functional expression of CCR9-thymus-expressed chemokine receptor-ligand pairs as well as the effects of IL-2 and IL-4, which may be especially important in cytokine/chemokine environment for the pathophysiological events of T-ALL CD4+ T-cell trafficking. PMID- 14559840 TI - Identification and functional analysis of tumor-infiltrating plasmacytoid dendritic cells in head and neck cancer. AB - The antitumor activity of IFN-alpha is well established. However, the role of the plasmacytoid dendritic cell (PDC), the major producer of IFN-alpha upon viral infection, in tumor biology is unknown. We sought to study the presence and function of PDC in a human solid tumor. Here, we demonstrate that PDCs infiltrate tumor tissue of patients with head and neck squamous cell carcinoma (HNSCC). Functional activity of PDC was examined by using CpG motif containing oligonucleotides, a defined microbial stimulus for PDCs (recognized via toll-like receptor 9). We found that HNSCC diminished the ability of PDC to produce IFN alpha in response to CpG motif containing oligonucleotide. Tumor-induced down regulation of toll-like receptor 9 was identified as one mechanism likely contributing to impaired PDC function within the tumor environment. In tumor draining lymph nodes, suppression of CpG-induced IFN-alpha production was less pronounced than in single-cell suspensions of primary tumor tissue. In these lymph nodes, CpG-induced IFN-alpha production was associated with increased levels of interferon-induced protein 10 and IFN-gamma and activation of CD4 and CD8 T cells. These results show for the first time the presence of PDCs in human solid tumor tissue and that tumors suppress the capacity of PDCs to produce IFN alpha. PDCs, which in the absence of appropriate stimulation are reported to promote regulatory CD8 T cells, may contribute to an impaired T-cell-mediated immune response in HNSCC. PMID- 14559841 TI - Human epithelial cancers secrete immunoglobulin g with unidentified specificity to promote growth and survival of tumor cells. AB - Immunoglobulins (Igs) are found thus far only to be produced by differentiated B lymphocytes. By immunohistochemistry analysis, in situ hybridization, and laser capture microdissection-assisted single-cell PCR, we demonstrate that human cancers of epithelial origin, including carcinomas of breast, colon, liver, lung, established epithelial cancer lines, as well as some normal lung tissues, also produce IgG in both cytoplasmic and secreted forms. Furthermore, blockade of tumor-derived IgG by either antisense DNA or antihuman IgG antibody increased programmed cell death and inhibited growth of cancer cells in vitro. More importantly, administration of antihuman IgG antibody also suppressed the growth of an IgG-secreting carcinoma line in immunodeficient nude mice. Our results support a role of tumor-derived IgG as growth factor for epithelial cancers. Prevalent expression of IgG in human carcinomas and its growth-promoting functions may have important implications in growth regulation and targeted therapy of human cancers. PMID- 14559842 TI - Activation of tumor-specific T lymphocytes by radio-frequency ablation of the VX2 hepatoma in rabbits. AB - Radio-frequency ablation (RFA) is used as a minimally invasive treatment for inoperable hepatic tumors. Immunological reactions secondary to RFA may play a role in the observed tumor control. In our study, the VX2 carcinoma was implanted into the liver of rabbits. After 3 weeks, tumors were treated with RFA or were left untreated. Peripheral blood lymphocytes were harvested before tumor implantation, 2 weeks postoperatively and at 2-week intervals thereafter. T cells were stimulated with lysates of either tumor tissue or nontumorous liver loaded on autologous antigen-presenting cells and their stimulation index was determined by [(3)H]thymidine incorporation. A 3-fold increase over background or controls was considered significant. Stimulation with phytohemagglutinin served as a positive control. The animals were necropsied, and liver and tumor tissue were analyzed immunohistologically for T-cell infiltration. T cells from tumor-bearing (n = 9) and RFA-treated (n = 11) animals were investigated in a follow-up study. The mean postoperative observation was 45 days. All of the 11 RFA-treated animals exhibited circulating T cells activated specifically toward tumor antigens throughout the observation period, which was accompanied by dense T-cell infiltration. In contrast, T cells of untreated tumor-bearing rabbits showed no reaction and only sparse T cell infiltration. We concluded that RFA induces a tumor-specific T-cell reaction in the otherwise unreactive tumor-bearing host, apparently overcoming immune tolerance and leading to the presentation of otherwise cryptic tumor antigens. Therefore, in addition to destroying tumor tissue, RFA induces an immune response against tumor antigens that may be exploited in multimodal antitumor strategies. PMID- 14559843 TI - B7-H1 blockade augments adoptive T-cell immunotherapy for squamous cell carcinoma. AB - In this report, we demonstrate that B7-H1, a B7 family molecule implicated in tumor immune evasion, is constitutively expressed on 66% of freshly isolated squamous cell carcinomas of the head and neck (SCCHN). To define the potential impact of tumor-associated B7-H1 on immunotherapy, the B7-H1-negative mouse SCC line, SCCVII, was transfected to express B7-H1. Although all of the animals succumbed to B7-H1/SCCVII tumors even after adoptive T-cell immunotherapy, the infusion of B7-H1 blocking monoclonal antibody with activated T cells cured 60% of animals. These data support B7-H1 blockade as a new approach to enhance the efficacy of T-cell immunotherapy. PMID- 14559844 TI - The alternative open reading frame of LAGE-1 gives rise to multiple promiscuous HLA-DR-restricted epitopes recognized by T-helper 1-type tumor-reactive CD4+ T cells. AB - The NY-ESO-1 and LAGE-1 genes are expressed by many human cancers, but not by normal tissues, with the exception of testis and placenta. The NY-ESO-1 and LAGE 1 genes give rise to multiple MHC class I and class II-presented epitopes derived from the open reading frames (ORF) 1 and 2. Here, we have investigated whether NY ESO-1/LAGE-1 ORF2 encodes promiscuous MHC class II-restricted epitopes. Using a set of overlapping peptides from the ORF2 protein sequence and autologous dendritic cells (DCs) from normal donors and melanoma patients, we have identified three HLA-DRB1*0401-restricted peptide sequences from the LAGE-1 ORF2 that are capable of stimulating T-helper 1-type melanoma-reactive CD4+ T cells. From these bulk CD4+ T cells, we have generated CD4+ T-cell clones able to recognize not only peptide-pulsed DCs but also autologous DCs loaded with the LAGE-1 ORF2 protein. We have demonstrated that these peptides not only bind to multiple HLA-DR molecules apart from HLA-DRB1*0401 but also stimulate CD4+ T cells when presented in the context of these HLA-DR molecules. Furthermore, our binding data have delineated two additional sequences capable of broadly binding to multiple HLA-DR molecules. Altogether, these data support the immunogenicity of NY-ESO-1/LAGE-1 ORF2 gene products and clearly demonstrate their capability to stimulate T-helper 1 type CD4+ T cells. Because of the role of these cells in promoting long-lasting antitumor CTL responses, our data provide a rationale for cancer vaccine trials with peptides derived from the NY-ESO-1/LAGE-1 ORF2 for a large fraction of patients with NY-ESO-1/LAGE-1(+) tumors. PMID- 14559845 TI - Comparison of the selective estrogen receptor modulator arzoxifene (LY353381) with tamoxifen on tumor growth and biomarker expression in an MCF-7 human breast cancer xenograft model. AB - Arzoxifene (ARZ) is a selective estrogen receptor (ER) modulator with greater bioavailability than raloxifene which is being developed as treatment for breast cancer. We have used an in vivo model of hormone-sensitive breast cancer to study the growth-inhibitory and pharmacodynamic effects of ARZ in comparison with the most widely used antiestrogen, tamoxifen (TAM). We compared the abilities of ARZ and TAM to antagonize the estrogen (E2)-dependent growth of MCF-7 human breast cancer xenografts in oophorectomized athymic mice. At four different time points over 28 days, we studied their time-related pharmacodynamic effects on biomarkers of tumor growth (cell proliferation/death measured by Ki-67 and apoptosis scores), cell cycle activity (cyclin D1 and p27(kip1)), and hormone-regulated gene expression (ERalpha, progesterone receptor, and pS2). Although maximal growth inhibition was seen after E2 withdrawal, ARZ and TAM induced significant and similar inhibition of E2-stimulated tumor growth. Inhibition of growth was reflected by changes in the tumor growth index (ratio posttreatment/pretreatment Ki-67/apoptosis scores). ARZ and TAM resulted in a significant (P < 0.001) increase in ER expression and reduction in progesterone receptor expression, whereas changes in cyclin D1 score were inversely related to p27(kip1) score. A significant but delayed biological effect was observed with a 10-fold lower dose of ARZ. These results show that ARZ is an effective antagonist of E2-stimulated breast cancer growth with similar growth-inhibitory and pharmacodynamic effects to TAM in this model. PMID- 14559846 TI - Letrozole inhibits tumor proliferation more effectively than tamoxifen independent of HER1/2 expression status. AB - BACKGROUND: The biological basis for the superior efficacy of neoadjuvant letrozole versus tamoxifen for postmenopausal women with estrogen receptor (ER) positive locally advanced breast cancer was investigated by analyzing tumor proliferation and expression of estrogen-regulated genes before and after the initiation of therapy. METHODS: Tumor samples were obtained at baseline and at the end of treatment from 185 patients participating in a double blind randomized Phase III study of neoadjuvant endocrine therapy. These paired specimens were simultaneously analyzed for Ki67, ER, progesterone receptor (PgR), trefoil factor 1 (PS2), HER1 (epidermal growth factor receptor), and HER2 (ErbB2 or neu) by semiquantitative immunohistochemistry. RESULTS: The treatment-induced reduction in geometric mean Ki67 was significantly greater with letrozole (87%) than tamoxifen (75%; analysis of covariance P = 0.0009). Differences in the average Ki67 reduction were particularly marked for ER-positive tumors that overexpressed HER1 and/or HER2 (88 versus 45%, respectively; P = 0.0018). Twenty-three of 92 tumors (25%) on tamoxifen and 14 of 93 on letrozole (15%) showed a paradoxical increase in Ki67 with treatment, and the majority of these cases was HER1/2 negative. Letrozole, but not tamoxifen, significantly reduced expression of the estrogen-regulated proteins PgR and trefoil factor 1, regardless of HER1/2 status (P < 0.0001). ER down-regulation occurred with both agents, although levels decreased more with tamoxifen (P < 0.0001). CONCLUSION: Letrozole inhibited tumor proliferation to a greater extent than tamoxifen. The molecular basis for this advantage appears complex but includes possible tamoxifen agonist effects on the cell cycle in both HER1/2+ and HER1/2- tumors. A pattern of continued proliferation despite appropriate down-regulation of PgR expression with estrogen deprivation or tamoxifen was also documented. This observation suggests the estrogenic regulation of proliferation and PgR expression may be dissociated in endocrine therapy resistant cells. PMID- 14559847 TI - Human cytochrome P450 3A7 has a distinct high catalytic activity for the 16alpha hydroxylation of estrone but not 17beta-estradiol. AB - Like catechol estrogens, 16alpha-hydroxylated estrogens are hormonally active, chemically reactive, and potentially mutagenic. We report here our novel findings that human CYP3A7 has a distinct high catalytic activity for the NADPH-dependent 16alpha-hydroxylation of estrone (E(1); at 10 nM to 200 microM substrate concentrations) but not for the 16alpha-hydroxylation of 17beta-estradiol (E(2)). At a physiologically relevant low substrate concentration (10 nM), CYP3A7 had a strong catalytic activity for the 16alpha-hydroxylation of E(1), and the ratio of its 16alpha-hydroxylation to 2-hydroxylation was 107%. In addition to 16alpha hydroxylation, CYP3A7 also had catalytic activity for the 2-, 4-, 6beta-, and 16beta-hydroxylation of E(1). However, when E(2) was the substrate, CYP3A7 had only very weak catalytic activity for its 16alpha-hydroxylation (<6% of E(1) 16alpha-hydroxylation), and the ratio of its 16alpha-hydroxylation to 2 hydroxylation was 10-33%. Enzyme kinetic analysis showed that the maximal velocity and substrate-binding affinity (1/K(m)) for CYP3A7-mediated 16alpha hydroxylation of E(1) were both approximately 10 times higher than those for E(2), thereby giving the maximal velocity:K(m) ratio of >100 times higher for the 16alpha-hydroxylation of E(1) than for E(2). Given the recent findings that human CYP3A7 is a polymorphic isoform also expressed in adult liver and certain extrahepatic tissues (in addition to fetal tissues), our data raise the possibility that CYP3A7 may be an important catalyst for the local and/or systemic formation of the procarcinogenic 16alpha-hydroxyestrone in women. PMID- 14559848 TI - Increased expression and activation of gelatinolytic matrix metalloproteinases is associated with the progression and recurrence of human cervical cancer. AB - Cancer-derived matrix metalloproteinases (MMPs) are proposed to be essential for tumor stromal invasion and subsequent metastasis. To explore the role of MMPs in cancer progression, we examined the expression of various MMPs and tissue inhibitors of MMPs in precancerous and cancerous lesions of the uterine cervix. Immunohistochemical studies demonstrated that MMP-2 and MMP-9 were expressed in >90% of squamous cell carcinomas (SCC) and 83-100% of high-grade squamous intraepithelial lesions (HSIL), but were less frequently expressed in low-grade squamous intraepithelial lesions and normal squamous epithelium (13%). MMP-1, MMP 14, and MMP-15 were detected in 55-81% of SCC cases, and MMP-1 was detected in 39% of HSIL. The tissue inhibitors of MMPs were weakly expressed in SCC (10-61%). By direct analysis of enzyme activities in microdissected specimens, we found that the gelatinolytic activity of MMP-9 was significantly higher in HSIL and SCC than in normal cervix (P < 0.01). The levels of active-form MMP-2 increased progressively from HSIL to SCC of stage I and more advanced stages (P < 0.01). The gelatinolytic activity of MMP-9 and active-form MMP-2 in SCC were strongly correlated with lymphovascular permeation and subsequent lymph node metastasis (P < 0.02). Moreover, the gelatinolytic activity and immunoreactive percentage of both MMP-2 and MMP-9 were significantly higher in SCC cases who had a recurrence than in those who remained free of disease (P < 0.001). Thus, our data demonstrate progressively up-regulated expression of MMP-2 and MMP-9 with SCC progression, and significant associations among their gelatinolytic activity and stage, nodal metastasis, and recurrence. PMID- 14559849 TI - The usefulness of serum human kallikrein 11 for discriminating between prostate cancer and benign prostatic hyperplasia. AB - Prostate-specific antigen (PSA) is the most useful tumor marker for diagnosis and monitoring of prostate cancer (CaP). Recently, we developed a specific immunoassay for human kallikrein 11 (hK11), one of the kallikrein gene family members, and found that hK11 was highly expressed in prostatic tissue and could be detected in seminal plasma (E. P. Diamandis et al., Cancer Res., 62: 295-300, 2002). The aim of this study was to investigate whether serum hK11 levels could be used to discriminate CaP from benign prostatic hyperplasia (BPH). We analyzed for hK11, total PSA, and percentage of free PSA, 150 serum samples from men with histologically confirmed BPH (n = 64) or CaP (n = 86). Total and free PSA levels were measured by the Immulite PSA assay, and hK11 levels were measured by our previously published immunofluorometric assay. Serum hK11 levels and the hK11:total PSA ratio were both significantly lower in CaP patients than in BPH patients. In the subgroup of patients with percentage of free PSA less than 20, an additional 54% of BPH patients could have avoided biopsies by using the hK11:total PSA ratio. Receiver operating characteristic (ROC) curve analysis demonstrated that the hK11:total PSA ratio [area under the curve (AUC), 0.83] and percentage of free PSA (AUC, 0.83) were much stronger predictors of CaP than total PSA (AUC, 0.69). These preliminary data suggest that the hK11:total PSA ratio could be a useful tumor marker for CaP and could be combined with percentage of PSA to further reduce the number of unnecessary prostatic biopsies. PMID- 14559851 TI - Lung cancer mortality in relation to age, duration of smoking, and daily cigarette consumption: results from Cancer Prevention Study II. AB - The magnitude of the effect of smoking duration on lung cancer mortality relative to that of intensity (cigarettes/day) has practical implications for both tobacco control policy and research. This issue was addressed by R. Doll and R. Peto (J. Epidemiol. Commun. Health, 32: 303-313, 1978) in their historic analysis of one of the few large cohort studies in which intensity and duration were estimated separately. Their findings have been interpreted to mean that smoking duration is much more important than smoking intensity in causing lung cancer. The separate contributions of smoking duration and intensity to lung cancer risk have not been evaluated in other large prospective studies. We studied participants in the Cancer Prevention Study II, followed from 1982 through 1988. After restricting to people 40-79 years old who smoked < or =40 cigarettes per day at enrollment, we fit Poisson models for four age groups and evaluated lung cancer mortality (M) in relation to smoking duration (D) and intensity (I) on a double-log scale, as suggested by the Armitage-Doll multistage carcinogenesis model. The age-specific mortality estimates for men (M(m)) and for women (M(w)), when transformed to the original scale, were: ages 40-49: M(m) = e(-17.9) x D(1.9) x I(0.95), M(w) = e( 20.2) x D(2.8) x I(0.96); ages 50-59: M(m) = e(-17.4) x D(2.6) x I(0.52), M(m) = e(-17.2) x D(2.2) x I(0.75); ages 60-69: M(m) = e(-15.7) x D(2.4) x I(0.37), M(m) = e(-14.1) x D(1.5) x I(0.78); ages 70-79: M(m) = e(-13.0) x D(1.8) x I(0.39), M(m) = e(-13.2) x D(1.3) x I(0.95). Our study confirms that years of cigarette smoking is far more important than the number of cigarettes smoked per day in predicting lung cancer risk in United States men, regardless of age, and provides new evidence that a qualitatively similar pattern holds for women. The results support measures to prevent the uptake of smoking by adolescents and increase cessation. We discuss reasons why the associations we observe are lower than those reported by R. Doll and R. Peto (J. Epidemiol. Commun. Health, 32: 303-313, 1978). PMID- 14559850 TI - Protein kinase Cepsilon is linked to 12-O-tetradecanoylphorbol-13-acetate-induced tumor necrosis factor-alpha ectodomain shedding and the development of metastatic squamous cell carcinoma in protein kinase Cepsilon transgenic mice. AB - Protein kinase Cepsilon (PKCepsilon), a Ca(2+)-independent, phospholipid dependent serine/threonine kinase, is among the PKC isoforms expressed in mouse epidermis. We reported that FVB/N transgenic mice that overexpress ( approximately 18-fold) PKCepsilon protein in basal epidermal cells and cells of the hair follicle develop papilloma-independent metastatic squamous cell carcinoma (mSCC) elicited by 7,12-dimethylbenz(a)anthracene-initiation and 12-O tetradecanoylphorbol-13-acetate (TPA)-promotion protocol. We now present that PKCepsilon transgenic mice elicit elevated serum tumor necrosis factor (TNF)alpha levels during skin tumor promotion by TPA, and this increase may be linked to the development of mSCC. A single topical application of TPA (5 nmol) to the skin, as early as 2.5 h after treatment, resulted in a significant (P < 0.01) increase (2 fold) in epidermal TNFalpha and more than a 6-fold increase in ectodomain shedding of TNFalpha into the serum of PKCepsilon transgenic mice relative to their wild-type littermates. Furthermore, this TPA-stimulated TNFalpha shedding was proportional to the level of expression of PKCepsilon in the epidermis. Using the TNF-alpha converting enzyme (TACE) inhibitor, TAPI-1, TPA-stimulated TNFalpha shedding could be completely prevented in PKCepsilon transgenic mice and isolated keratinocytes. These results indicate that PKCepsilon signal transduction pathways to TPA-stimulated TNFalpha ectodomain shedding are mediated by TACE, a transmembrane metalloprotease. Using the superoxide dismutase mimetic CuDIPs and the glutathione reductase mimetic ebselen, TPA-stimulated TNFalpha shedding from PKCepsilon transgenic mice could be completely attenuated, implying the role of reactive oxygen species. Finally, i.p. injection of a TNFalpha synthesis inhibitor, pentoxifylline, during skin tumor promotion completely prevented the development of mSCC in PKCepsilon transgenic mice. Taken together, these results indicate that: (a) PKCepsilon activation is an initial signal in TPA-induced shedding of TNFalpha from epidermal keratinocytes; (b) PKCepsilon-mediated signals to TACE are possibly mediated through reactive oxygen species; and (c) TPA-induced TNFalpha shedding may play a role in the development of mSCC in PKCepsilon transgenic mice. PMID- 14559852 TI - Correspondence re: R. Varon et al., Mutations in the Nijmegen breakage syndrome gene (NBS1) in childhood acute lymphoblastic leukemia. Cancer Res., 61: 3570 3572, 2001. PMID- 14559854 TI - Correspondence re: L. Yang et al., Predominant suppression of apoptosome by inhibitor of apoptosis protein in non-small cell lung cancer H460 cells: therapeutic effect of a novel polyarginine-conjugated Smac peptide. Cancer Res., 63: 831-837, 2003. PMID- 14559856 TI - FK506 potentiates NGF-induced neurite outgrowth via the Ras/Raf/MAP kinase pathway. AB - Nerve growth factor (NGF) and other members of the neurotrophin family are critical for the survival and differentiation of neurons within the peripheral and central nervous systems. Neurophilin ligands, including FK506, potentiate NGF induced neurite outgrowth in several experimental models, although the mechanism of this potentiation is unclear. Therefore, we tested which signaling pathways were involved in FK506-potentiated neurite outgrowth in SH-SY5Y neuroblastoma cells using specific pharmacological inhibitors of various signaling molecules. Inhibitors of Ras (lovastatin), Raf (GW5074), or MAP kinase (PD98059 and U0126) blocked FK506 activity, as did inhibitors of phospholipase C (U73122) and phosphatidylinositol 3' kinase (LY294002). Protein kinase C inhibitors (Go6983 and Ro31-8220) slightly but significantly inhibited neurite outgrowth, whereas inhibitors of p38 MAPK (SB203580) or c-Jun N-terminal kinase (SP600125) had no effect. These data suggest that FK506 potentiates neurite outgrowth through the Ras/Raf/MAP kinase signaling pathway downstream of phospholipase C and phosphatidylinositol 3' kinase. PMID- 14559858 TI - Plasma membrane-associated nucleoside diphosphate kinase (nm23) in the heart is regulated by beta-adrenergic signaling. AB - 1. Receptor-independent activation of heterotrimeric G proteins by plasma membrane-associated nucleoside diphosphate kinase (NDPK) has been demonstrated in vivo, and elevated levels of NDPK were found in purified sarcolemmal membranes of patients with end-stage heart failure. 2. Among 22 consecutive patients with chronic heart failure who underwent cardiac transplantation, those treated with a beta-blocker (n=8) had a 65% lower NDPK content and activity in the cardiac sarcolemma, compared to patients with similar base line characteristics who had no beta-blocker therapy (n=14). 3. The lower NDPK was associated with a reduced NDPK-dependent, Gi-mediated inhibition of adenylyl cyclase activity, as assessed by in vitro measurement of adenylyl cyclase activity in the presence of GDP or its kinase-resistant analog guanosine 5'-O-(2-thio)diphosphate (GDPbetaS). 4. We further tested whether treatment with a beta-adrenergic agonist would induce an increase in sarcolemmal NDPK. Rats treated with isoproterenol developed myocardial hypertrophy, and NDPK in the sarcolemma rose by 60% during 14 days of treatment. The beta-blocker propranolol prevented both effects. When hypertrophy was induced with thyroid hormone, NDPK did not increase. 5. In conclusion, chronic activation of beta-adrenergic receptors increases the binding of NDPK to cardiac sarcolemma, where it may activate heterotrimeric G proteins. PMID- 14559857 TI - Effect of two phenanthrene alkaloids on angiotensin II-induced leukocyte endothelial cell interactions in vivo. AB - 1. The present study has evaluated the effect of two phenanthrene alkaloids, uvariopsine and stephenanthrine, on angiotensin II (Ang-II)-induced leukocyte endothelial cell interactions in vivo and the mechanisms involved in their activity. Intravital microscopy within the rat mesenteric microcirculation was used. 2. A 60 min superfusion with 1 nm Ang-II induced a significant increase in the leukocyte-endothelial cell interactions that were completely inhibited by 1 microm uvariopsine cosuperfusion. A lower dose of 0.1 microm significantly reduced Ang-II-induced leukocyte adhesion by 75%. 3. When Ang-II was cosuperfused with 1 and 0.1 microm stephenanthrine, Ang-II-induced leukocyte responses were significantly diminished. A lower dose of 0.01 microm only affected Ang-II induced leukocyte adhesion. 4. Both alkaloids inhibited Ang-II-induced endothelial P-selectin upregulation and the generation of reactive oxygen species (ROS) in endothelial cells stimulated with Ang-II, in fMLP-stimulated human neutrophils (PMNs) and in the hypoxanthine-xanthine oxidase system. However, cyclic AMP levels in PMNs stimulated with fMLP were not affected. 5. Uvariopsine and stephenanthrine inhibited PAF-induced elevations in intracellular calcium levels in PMNs (IC50 values: 15.1 and 6.1 microm respectively) and blocked the binding of [3H]PAF to these leukocytes. They also reduced PAF-induced increases in intracellular levels of superoxide anion and hydrogen peroxide. 6. In conclusion, stephenanthrine and uvariopsine are potent inhibitors of Ang-II induced leukocyte accumulation in vivo. This effect appears to be mediated through ROS scavenging activity and blockade of PAF receptor. Thus, they have potential therapeutic interest for the control of leukocyte recruitment that occurs in cardiovascular disease states in which Ang-II is involved. PMID- 14559860 TI - Practice patterns of sentinel node biopsy at five comprehensive cancer centers. PMID- 14559859 TI - Hypoxia-inducible factor-1 and activator protein-1 modulate the upregulation of CYP3A6 induced by hypoxia. AB - 1. Moderate hypoxia in vivo and serum from rabbits subjected to moderate hypoxia (SHYPO) in vitro reduce CYP1A1 and 1A2 p450 isoforms and upregulate CYP3A6. The aim of this project was to investigate the signal transduction pathways implicated in the upregulation of CYP3A6 expression by hypoxia. 2. Hypoxia in vivo and SHYPO in vitro increased the expression of hypoxia-inducible factor 1alpha (HIF-1alpha) and c-jun, as well as CYP3A6. By electrophoresis mobility shift assay, it was shown that HIF-1 and activator protein-1 (AP-1) bind to CYP3A6 oligonucleotide probe after exposure to hypoxia in vivo and SHYPO in vitro. The effects of hypoxia in vivo or SHYPO in vitro were reproduced by CoCl2 and lead acetate, activators of HIF-1 and AP-1, respectively. 2. PD98059, a p42/44 MAPK inhibitor, prevented the increase of CYP3A6 and c-jun, but did not impede the increase of HIF-1alpha and binding to CYP3A6 oligonucleotide probe. Genistein, an inhibitor of protein tyrosine kinases (PTKs), prevented the increase in HIF-1alpha, c-jun and CYP3A6, as well as HIF-1 and AP-1 binding to CYP3A6 oligonucleotide probe. Moreover, hypoxia in vivo induced constitutive androstane receptor (CAR) as well as CAR binding to the CYP3A6 oligonucleotide probe, but not the pregnane X receptor. 4. In conclusion, hypoxia in vivo and SHYPO induce the expression of CYP3A6. The in vitro induction of CYP3A6 by SHYPO is PTK- and p42/44 MAPK-dependent. The present data support the hypothesis that HIF-1 and AP-1 are part of the signalling pathway leading to CYP3A6 induction by hypoxia. PMID- 14559861 TI - Setting goals for cancer mortality: earlier initiatives offer lessons for the future. PMID- 14559862 TI - Estimating reductions in cancer mortality. PMID- 14559863 TI - New charity chief discusses his fast-paced agenda. PMID- 14559864 TI - Acceptance of alternative therapies slowly growing in United kingdom. PMID- 14559865 TI - Bristol Cancer Help Center paving the way for CAM therapy. PMID- 14559866 TI - Large-scale science: Boston area institutions given 100 million dollars for genomics institute. PMID- 14559867 TI - Stat bite: Incidence of and mortality from childhood cancers, 1975-2000. PMID- 14559868 TI - Long-term effects of childhood cancer need to be documented, board says. PMID- 14559871 TI - Mammography screening: are women really giving informed consent? (Counterpoint). PMID- 14559870 TI - Mammography screening: are women really giving informed consent? PMID- 14559872 TI - Mammography screening: are women really giving informed consent? (Countering the counterpoint). PMID- 14559873 TI - Emergence of sentinel node biopsy in breast cancer as standard-of-care in academic comprehensive cancer centers. AB - BACKGROUND: Ongoing clinical trials are addressing the accuracy and safety of sentinel node biopsy (SNB) in the treatment of breast cancer; however, SNB is already increasingly being used in clinical practice. This study examined the extent and time trends of the use of SNB in stage I and II breast cancer patients. METHODS: Clinical data were collected from stage I and II (tumor size < or =5.0 cm) breast cancer patients (n = 3003) who were treated at five comprehensive cancer centers between July 1, 1997, and December 31, 2000. Axillary surgery was classified as SNB alone, SNB + axillary node dissection (AND), AND alone, or none. Patterns of use of axillary surgery were summarized as the percentage of patients receiving each surgery type. The statistical significance of time trends for the use of SNB alone was analyzed by logistic regression models. All statistical tests were two-sided. RESULTS: Overall, SNB alone was used in 13% of patients, SNB + AND in 22%, AND alone in 59%, and no axillary surgery in 6%. Use of SNB alone was statistically significantly associated with breast-conserving surgery of both smaller (< or =2 cm) and larger tumors (2-5 cm) (P<.001 for both associations). For stage I cancer patients treated with breast-conserving surgery (n = 1763), use of SNB increased statistically significantly over the study period, from 8% in 1997 to 9%, 14%, 15%, 22%, 42%, and 58% for the next six consecutive 6-month time intervals, respectively. After controlling for center, age, and comorbidity, the odds ratio for the use of SNB alone was 2.30 (95% confidence interval = 1.88 to 2.82) for each 6-month interval (P<.001). CONCLUSIONS: Widespread use of SNB outside the clinical trial setting suggests that oncologists at cancer centers in our study have accepted SNB as standard-of-care for the treatment of breast cancer. This acceptance, if it occurs in other cancer centers and community practice, may affect accrual and generalizability of ongoing clinical trials of SNB. PMID- 14559875 TI - A prospective study of pigmentation, sun exposure, and risk of cutaneous malignant melanoma in women. AB - BACKGROUND: Although sun exposure is an established cause of cutaneous malignant melanoma, possible interactions with host factors remain incompletely understood. Here we report the first results from a large prospective cohort study of pigmentation factors and sun exposure in relation to melanoma risk. METHODS: The Women's Lifestyle and Health Cohort Study included 106 379 women from Norway and Sweden who were aged 30-50 years in 1991 or 1992 when they completed an extensive questionnaire on personal characteristics and exposures. Linkages to national registries ensured complete follow-up through December 31, 1999. Poisson regression models were used to estimate relative risks (RRs). All statistical tests were two-sided. RESULTS: During an average follow-up of 8.1 years, 187 cases of melanoma were diagnosed. Risk of melanoma was statistically significantly associated with increasing body surface area (RR for > or =1.79 m2 versus < or =1.61 m2 = 1.60, 95% confidence interval [CI] = 1.03 to 2.48; P(trend) =.02), number of large asymmetric nevi on the legs (RR for > or =7 nevi versus 0 nevi = 5.29, 95% CI = 2.33 to 12.01; P(trend)<.001), hair color (RR for red versus dark brown or black = 4.05, 95% CI = 2.11 to 7.76; P(trend)<.001), sunburns per year at ages 10-19, 20-29, and 30-39 years (P(trend)<.001, P(trend) =.03, and P(trend) =.05, respectively), and use of a device that emits artificial light (solarium) one or more times per month (P =.04). CONCLUSIONS: Our results confirm previous findings that hair color, number of nevi on the legs, and history of sunburn are risk factors for melanoma and suggest that use of a solarium is also associated with melanoma risk. Adolescence and early adulthood appear to be among the most sensitive age periods for the effects of sunburn and solarium use on melanoma risk. However, it may be too early to see the full effect of adult exposures in this cohort. PMID- 14559874 TI - Lack of serologic evidence for prevalent simian virus 40 infection in humans. AB - BACKGROUND: Propagation of poliovirus in monkey kidney cells led to the inadvertent contamination of poliovirus vaccines with simian virus 40 (SV40) between 1955 and 1963. Recent studies using polymerase chain reaction-based strategies have detected SV40 DNA in a large number of tumor types. The finding of SV40 DNA in tumors from individuals who are too young to have been exposed to SV40-contaminated vaccines has led to the suggestion that SV40 has become a prevalent transmissible human pathogen. To test this hypothesis, we screened human sera for antibodies to SV40 using direct and competitive enzyme-linked immunosorbent assays (ELISAs). METHODS: An ELISA was developed using recombinant SV40 virus-like particles (VLPs) and was validated using sera from naturally infected macaques. VLPs of SV40 and the related ubiquitous human polyomaviruses, JCV and BKV, were used to screen human sera to determine the prevalence of SV40, JCV, and BKV antibodies among a normal population of control subjects (n = 487) and among case patients with either osteosarcoma (n = 122) or prostate cancer (n = 90). A competitive ELISA in which sera were pre-adsorbed with each type of VLP was used to identify cross-reactive antibodies. Correlations of reactivity among the three polyomavirus types were calculated using the Spearman correlation coefficient. All statistical tests were two-sided. RESULTS: BKV and JCV antibodies were prevalent in all case patients and control subjects examined. In contrast, only 6.6% (46/699) of serum samples were positive for SV40 antibodies by ELISA; however, none of these samples could be confirmed as having authentic SV40 antibodies following pre-adsorption with JCV or BKV VLPs. CONCLUSION: These data indicate that some individuals have BKV and/or JCV antibodies that cross react with SV40, but they do not provide support for SV40 being a prevalent human pathogen. PMID- 14559876 TI - Age- and sex-specific incidence of childhood leukemia by immunophenotype in the Nordic countries. AB - BACKGROUND: Studies from various countries have found an increasing incidence of childhood leukemia in recent decades. To characterize time trends in the age- and sex-specific incidence of childhood acute leukemia during the last 20 years in the Nordic countries, we analyzed a large set of population-based data from the Nordic Society of Paediatric Haematology and Oncology (NOPHO) in their acute leukemia database covering a population of approximately 5 million children aged 0-14 years. METHODS: Temporal trends in acute myeloid leukemia and acute lymphoblastic leukemia incidence rates overall and for acute lymphoblastic leukemia immunophenotypes and for specific age groups were analyzed by Poisson regression adjusting for age, sex, and country. All statistical tests were two sided. RESULTS: We identified 1595 girls and 1859 boys diagnosed with acute lymphoblastic leukemia between January 1, 1982, and December 31, 2001, and 260 girls and 224 boys diagnosed with de novo acute myeloid leukemia between January 1, 1985, and December 31, 2001. No statistically significant change was seen in the overall incidence rate for acute lymphoblastic leukemia during the 20-year study (annual change = 0.22%, 95% confidence interval [CI] = -0.36% to 0.80%). The incidence rate of B-precursor acute lymphoblastic leukemia remained unchanged (annual change = 0.30%, 95% CI = -0.57% to 1.18%) from January 1, 1986, through December 31, 2001. A somewhat lower incidence in the first years of the study period indicated an early increasing incidence of B-precursor acute lymphoblastic leukemia that corresponded to a simultaneous decreasing incidence of unclassified acute lymphoblastic leukemia. Incidences of T-cell acute lymphoblastic leukemia (annual change = 1.55%, 95% CI = -1.14% to 4.31%) and acute myeloid leukemia (annual change = 0.58%, 95% CI = -1.24% to 2.44%) were stable during the study period. CONCLUSION: Incidences of acute myeloid leukemia overall, acute lymphoblastic leukemia overall, and specific acute lymphoblastic leukemia immunophenotypes have been stable in the Nordic countries over the past two decades. PMID- 14559877 TI - Ethnic neutropenia and treatment delay in African American women undergoing chemotherapy for early-stage breast cancer. AB - Disparities in breast cancer survival have been observed between African American and white women. There are also known differences in mean baseline white blood cell (WBC) count among racial and ethnic groups. If the WBC count falls below conventionally defined treatment thresholds for patients undergoing adjuvant chemotherapy, reduced doses or treatment delays may occur, which could lead to race-based differences in treatment duration. We used the tumor registry at Columbia-Presbyterian Medical Center to identify 1178 women with newly diagnosed stage I and II breast cancer from whom we collected base-line information for 73 African American women and 126 age- and tumor stage-matched white women. Of these women, 43 African American and 93 white women underwent adjuvant chemotherapy. African American women had statistically significantly lower WBC counts than white women at diagnosis (6.2 x 10(9)/L for African American women versus 7.4 x 10(9)/L for white women, difference = 1.2, 95% confidence interval [CI] = 0.2 to 1.2; P =.02) and after treatment (5.3 x 10(9)/L for African American women versus 6.4 x 10(9)/L for white women, difference = 1.1, 95% CI = 0.2 to 2.5; P =.03). Overall, African American women required a statistically significantly longer duration of treatment than white women (19 weeks versus 15 weeks, respectively, difference = 4 weeks, 95% CI = 0.5 to 7.2 weeks; P =.03). The lower baseline WBC counts and longer duration of treatment for early-stage breast cancer in African American women compared with those in white women result in lower dose intensity of treatment for African American women, possibly contributing to observed racial differences in breast cancer survival. PMID- 14559878 TI - Shared genetic susceptibility to breast cancer, brain tumors, and Fanconi anemia. AB - Fanconi anemia is an inherited disease characterized by bone marrow failure, congenital malformations, and predisposition to cancer. The breast cancer susceptibility gene BRCA2 was recently found to be associated with Fanconi anemia complementation group D1 (FA-D1). We examined four kindreds afflicted with Fanconi anemia for the presence of germline BRCA2 mutations. One kindred, of Ashkenazi Jewish ancestry, had five members who were diagnosed with breast cancer and two cousins who were BRCA2*6174delT/C3069X compound heterozygotes and had Fanconi anemia and brain tumors. In another kindred of Ashkenazi Jewish and Lithuanian Catholic ancestry, a child with Fanconi anemia and a medulloblastoma was a BRCA2*6174delT/886delGT compound heterozygote. Two other kindreds each contained a Fanconi anemia-afflicted child who developed medulloblastoma; one child was of Latin American ancestry and a compound heterozygote for BRCA2*I2490T/ 5301insA and the other was African American and a compound heterozygote for BRCA2*Q3066X/E1308X. Median age of the Fanconi anemia-afflicted children at brain tumor diagnosis was 3.5 years. The co-occurrence of brain tumors, Fanconi anemia, and breast cancer observed in one of these kindreds constitutes a new syndromic association. Individuals who carry a germline BRCA2 mutation and who plan to have children with a partner of Ashkenazi Jewish descent should consider undergoing genetic counseling. PMID- 14559879 TI - Re: Cancer incidence in Denmark following exposure to poliovirus vaccine contaminated with simian virus 40. PMID- 14559880 TI - Re: Roles of radiation dose, chemotherapy, and hormonal factors in breast cancer following Hodgkin's disease. PMID- 14559882 TI - Recurrent episodes of involuntary masticatory spasms induced by continuous infusion of oxaliplatin. PMID- 14559884 TI - Re: Zinc supplement use and risk of prostate cancer. PMID- 14559885 TI - Predictors of occult metastasis in clinical stage I nonseminoma: a systematic review. AB - PURPOSE: Patients with clinical stage I nonseminomatous testicular germ cell tumor should ideally receive adjuvant therapy only when they are at high risk for occult metastasis. We aimed to quantify the importance of predictors for occult metastasis by performing a systematic review of the relevant literature. In addition, we reviewed published multivariable models and risk-adapted treatment policies. PATIENTS AND METHODS: We identified 23 publications between 1979 and 2001, reporting a total of 2,587 patients. Twenty-nine percent of the patients (759 of 2,587 patients) had occult metastases, which was diagnosed either at retroperitoneal lymph node dissection (n = 193) or during follow-up (n = 566). Odds ratios (OR) were pooled using meta-analysis techniques. RESULTS: The presence of vascular invasion of the primary tumor cells had the strongest effect (OR, 5.2; 95% CI, 4.0 to 6.8). Immunohistochemical staining of the primary tumor cells with the MIB-1 monoclonal antibody showing proliferative activity was a promising predictor (OR, 4.7; 95% CI, 2.0 to 11). Intermediate effects were found for embryonal carcinoma in the primary tumor (OR, 2.9; 95% CI, 2.0 to 4.4) and a high pathologic stage of the tumor (OR, 2.6; 95% CI, 1.8 to 3.8). Size of the primary tumor and age of the patient had weaker though also statistically significant associations with occult metastasis. Until now, multivariable models often included vascular invasion and embryonal carcinoma with one or two weaker predictors. None of the published risk-adapted treatment policies included MIB-1 staining. CONCLUSION: Several strong predictors for occult metastasis were identified. A risk-adapted treatment policy should be developed that incorporates all relevant predictors so that adjuvant therapy is targeted better to those with occult metastases. PMID- 14559886 TI - The oral neurokinin-1 antagonist aprepitant for the prevention of chemotherapy induced nausea and vomiting: a multinational, randomized, double-blind, placebo controlled trial in patients receiving high-dose cisplatin--the Aprepitant Protocol 052 Study Group. AB - PURPOSE: In early clinical trials with patients receiving highly emetogenic chemotherapy, the neurokinin antagonist aprepitant significantly enhanced the efficacy of a standard antiemetic regimen consisting of a type-three 5 hydroxytryptamine antagonist and a corticosteroid. This multicenter, randomized, double-blind, placebo-controlled phase III study was performed to establish definitively the superiority of the aprepitant regimen versus standard therapy in the prevention of chemotherapy-induced nausea and vomiting (CINV). PATIENTS AND METHODS: Patients receiving cisplatin > or = 70 mg/m2 for the first time were given either standard therapy (ondansetron and dexamethasone on day 1; dexamethasone on days 2 to 4) or an aprepitant regimen (aprepitant plus ondansetron and dexamethasone on day 1; aprepitant and dexamethasone on days 2 to 3; dexamethasone on day 4). Patients recorded nausea and vomiting episodes in a diary. The primary end point was complete response (no emesis and no rescue therapy) on days 1 to 5 postcisplatin, analyzed by a modified intent-to-treat approach. Treatment comparisons were made using logistic regression models. Tolerability was assessed by reported adverse events and physical and laboratory assessments. RESULTS: The percentage of patients with complete response on days 1 to 5 was significantly higher in the aprepitant group (72.7% [n = 260] v 52.3% in the standard therapy group [n = 260]), as were the percentages on day 1, and especially on days 2 to 5 (P <.001 for all three comparisons). CONCLUSION: Compared with standard dual therapy, addition of aprepitant was generally well tolerated and provided consistently superior protection against CINV in patients receiving highly emetogenic cisplatin-based chemotherapy. PMID- 14559887 TI - New prognostic factors for stage I testicular cancer: but will they make it to broadway? PMID- 14559888 TI - Why do we need another antiemetic? Just ask. PMID- 14559889 TI - The costs of conducting clinical research. AB - PURPOSE: Physicians frequently receive payment for enrolling subjects onto clinical trials. Some view these payments as conflicts of interest. Others contend that these payments are necessary reimbursements for conducting clinical research. We evaluated the clinical and nonclinical hours and costs associated with conducting a mock phase III clinical research trial. METHODS: We collected data from representatives of 21 clinical sites, on the numbers of hours associated with 13 activities necessary to the conduct of clinical research. The hours were based on enrolling 20 patients in a 12-month randomized placebo controlled trial of a new chemotherapeutic agent. The outcome measures were disease progression and quality-of-life reports. These costs were evaluated for both government and pharmaceutical industry-sponsored trials. RESULTS: On average, 4,012 hours (range, 1,512 to 13,319 hours) were required for a government-sponsored trial, and 3,998 hours (range: 1735 to 15,699) were required for a pharmaceutical industry-sponsored trial involving 20 subjects with 17 office visits, or approximately 200 hours per subject. Thirty-two percent of the hours were devoted to nonclinical activities, such as institutional review board submission and completion of clinical reporting forms. On average, excluding overhead expenses, it cost slightly more than 6,094 dollars (range, 2,098 dollars to 19,285 dollars) per enrolled subject for an industry-sponsored trial, including 1,999 dollars devoted to nonclinical costs. CONCLUSION: Based on the results of our mock trial, the time required for nontreatment trial activities is considerable, and the associated costs are substantial. PMID- 14559890 TI - Researching the cost of research. PMID- 14559891 TI - Addition of the oral NK1 antagonist aprepitant to standard antiemetics provides protection against nausea and vomiting during multiple cycles of cisplatin-based chemotherapy. AB - PURPOSE: This analysis evaluated whether the antiemetic efficacy of the NK1 receptor antagonist aprepitant (EMEND trade mark, Merck, Whitehouse Station, NJ) plus standard antiemetics could be sustained for up to six cycles of cisplatin based chemotherapy. PATIENTS AND METHODS: Patients receiving cisplatin > or = 70 mg/m2 were blindly assigned to receive one of the following three regimens: (1) aprepitant 375 mg 1 hour before cisplatin on day 1 and aprepitant 250 mg on days 2 to 5 (n = 35); (2) aprepitant 125 mg before cisplatin and aprepitant 80 mg on days 2 to 5 (n = 81); or (3) placebo before cisplatin on days 2 to 5 (n = 86). All groups received ondansetron 32 mg and dexamethasone 20 mg before cisplatin, and dexamethasone 8 mg on days 2 to 5. The primary end point was complete response (no emesis and no rescue therapy) over 5 days following cisplatin in up to six cycles. A cumulative probability analysis using a model for transitional probabilities was used to analyze the data. The aprepitant 375/250-mg regimen was discontinued early in light of new pharmacokinetic data. RESULTS: In the first cycle, 64% of patients in the aprepitant group and 49% in the standard therapy group had a complete response. Thereafter, complete response rates for the aprepitant group were still 59% by cycle 6, but decreased to 34% by cycle 6 for the standard therapy group. Reasons for discontinuation were similar across treatment groups. CONCLUSION: Compared with patients who received standard therapy, those who received only the aprepitant regimen had better and more sustained protection against chemotherapy-induced nausea and vomiting over multiple cycles. PMID- 14559892 TI - The effect on tumor response of adding sequential preoperative docetaxel to preoperative doxorubicin and cyclophosphamide: preliminary results from National Surgical Adjuvant Breast and Bowel Project Protocol B-27. AB - PURPOSE: The National Surgical Adjuvant Breast and Bowel Project Protocol B-27 was designed to determine the effect of adding docetaxel after four cycles of preoperative doxorubicin and cyclophosphamide (AC) on clinical and pathological response rates and on disease-free and overall survival of women with operable breast cancer. PATIENTS AND METHODS: Women (N = 2,411) with operable primary breast cancer were randomly assigned to receive either four cycles of preoperative AC followed by surgery (group I), or four cycles of AC followed by four cycles of docetaxel, followed by surgery (group II), or four cycles of AC followed by surgery and then four cycles of docetaxel (group III). Clinical and pathologic tumor responses to preoperative therapy were assessed. RESULTS: Mean tumor size (4.5 cm) and other key characteristics were evenly balanced among the three treatment arms. Grade 4 toxicity was observed in 10.3% of 2,400 patients during AC treatment, and in 23.4% of 1584 patients during docetaxel treatment. Compared to preoperative AC alone, preoperative AC followed by docetaxel increased the clinical complete response rate (40.1% v 63.6%; P <.001), the overall clinical response rate (85.5% v 90.7%; P <.001), the pathologic complete response rate (13.7% v 26.1%; P <.001), and the proportion of patients with negative nodes (50.8% v 58.2%; P <.001). Pathologic primary breast tumor response was a significant predictor of pathologic nodal status (P <.001). CONCLUSION: The addition of four cycles of preoperative docetaxel after four cycles of preoperative AC significantly increased clinical and pathologic response rates for operable breast cancer. PMID- 14559893 TI - Molecular basis for synergistic transcriptional activation by Oct1 and Sox2 revealed from the solution structure of the 42-kDa Oct1.Sox2.Hoxb1-DNA ternary transcription factor complex. AB - The Oct and Sox transcription factors control many different aspects of neural development and embryogenesis, often binding to adjacent sites on DNA, and interacting with one another through their DNA binding domains to regulate transcription synergistically. Oct proteins contain two DNA binding domains (POUS and POUHD) connected by a flexible linker, which interact with DNA in a bipartite manner. Residual dipolar coupling measurements on the binary Oct1.DNA complex reveal that the two domains are characterized by distinct alignment tensors in both phage pf1 and polyethylene glycol/hexanol liquid crystalline media. We show that this difference is due to a fast microscopic dissociation/association process involving alternative binding modes for the weaker binding POUS domain in the binary complex. Upon binding of Sox2 to an adjacent site in the Hoxb1 regulatory element, all components of the ternary Oct1.Sox2.DNA complex share a single alignment tensor. Thus ternary complex formation increases the site specific affinity of Oct1 for DNA by effectively locking the POUS domain in a single orientation on the DNA. The solution NMR structure of the ternary 42 kDa Oct1.Sox2.Hoxb1-DNA complex, determined by novel procedures based on orientational restraints from dipolar couplings and conjoined rigid body/torsion angle dynamics, reveals that Sox2 and POUS interact through a predominantly hydrophobic interface, surrounded by a ring of electrostatic interactions. These observations suggest a mechanism of combinatorial control involving direct protein-protein interactions on the DNA whereby Oct1 in conjunction with a co interacting transcription factor provide cell-specific transcription regulation. PMID- 14559894 TI - Cytoskeletal reorganization dependence of signaling by the gonadotropin-releasing hormone receptor. AB - Activation of classical G protein-coupled receptors (GPCRs) like the mammalian gonadotropin-releasing hormone receptor (GnRHR) typically stimulates heterotrimeric G protein molecules that subsequently activate downstream effectors. Receptor activation of heterotrimeric G protein pathways primarily controls intermediary cell metabolism by elevation or diminution of soluble cytoplasmic second messenger molecules. We have demonstrated here that stimulation of the GnRHR also results in a dramatic change in both cell adhesion and superstructural morphology. Gonadotropin-releasing hormone (GnRH) receptor activation rapidly increases the capacity of HEK293 cells expressing the GnRHR to remain matrix-adherent in the face of fluid insults. Coinciding with this profound elevation in matrix adherence, we demonstrated a GnRH-induced alteration in both cell morphology and the de novo generation of polymerized actin structures. GnRH induction of cytoskeletal remodeling was correlated with significant increases in the tyrosine phosphorylation status of a series of cytoskeletal associated proteins, e.g. focal adhesion kinase (FAK), c-Src, and microtubule-associated protein kinase (MAPK or ERK1/2). The activation of the distal downstream effector ERK1/2 was demonstrated to be sensitive to the disrupters of cytoskeletal rearrangement, cytochalasin D and latrunculin B. In addition to the sensitivity of ERKs to cytoskeletal integrity, GnRH-induced FAK and c-Src kinase activation were sensitive to these agents and the fibronectin integrin antagonistic RGDS peptide. Activation of ERK was dependent on its protein-protein assembly with FAK and c-Src at focal adhesion complexes. Induction of the cell remodeling event leading to this signaling complex assembly occurred primarily via GnRHR activation of the monomeric G protein Rac but not RhoA. These findings demonstrated a clear divergence of GnRHR signaling via the Rac monomeric G protein focal adhesion signaling complex assembly and cytoskeletal remodeling independent of the classical heterotrimeric G protein controlled phospholipase C-beta pathway. PMID- 14559895 TI - Functional analysis of the transmembrane domain and activation cleavage of human corin: design and characterization of a soluble corin. AB - Corin is a cardiac transmembrane serine protease. In cell-based studies, corin converted pro-atrial natriuretic peptide (pro-ANP) to mature ANP, suggesting that corin is potentially the pro-ANP convertase. In this study, we evaluated the importance of the transmembrane domain and activation cleavage in human corin. We showed that a soluble corin that consists of only the extracellular domain was capable of processing recombinant human pro-ANP in cell-based assays. In contrast, a mutation at the conserved activation cleavage site, R801A, abolished the function of corin, demonstrating that the activation cleavage is essential for corin activity. These results allowed us to design, express, and purify a mutant soluble corin, EKsolCorin, that contains an enterokinase recognition sequence at the activation cleavage site. Purified EKsolCorin was activated by enterokinase in a dose-dependent manner. Activated EK-solCorin had hydrolytic activity toward peptide substrates with a preference for Arg and Lys residues in the P-1 position. This activity of EKsolCorin was inhibited by trypsin-like serine protease inhibitors but not inhibitors of chymotrypsin-like, cysteine-, or metallo-proteases. In pro-ANP processing assays, purified active EKsolCorin converted recombinant human pro-ANP to biologically active ANP in a highly sequence-specific manner. The pro-ANP processing activity of EKsolCorin was not inhibited by human plasma. Together, our data indicate that the transmembrane domain is not necessary for the biological activity of corin but may be a mechanism to localize corin at specific sites, whereas the proteolytic cleavage at the activation site is an essential step in controlling the activity of corin. PMID- 14559896 TI - Involvement of a chaperone regulator, Bcl2-associated athanogene-4, in apolipoprotein B mRNA editing. AB - Apobec-1 is the catalytic subunit of a multicomponent editosome complex that mediates apolipoprotein B (apoB) mRNA editing. We isolated a novel apobec-1 interacting protein by yeast two-hybrid cloning and identified the protein as BAG 4. BAG-4, a chaperone-regulating protein, also known as SODD (silencer of death domains), is a member of the BAG family of proteins. In this report, we found that apobec-1 is localized in the perinucleolar compartment in HepG2 cells and rat liver MCR-RH7777 cells. BAG-4 binds to apobec-1 via its N-terminal region independent of the BAG domain. It is ubiquitously expressed with predominant occurrence in human pancreas, heart, brain, and placenta. Immunoprecipitation experiments confirmed that BAG-4 interacts with Hsc70/Hsp90 in HepG2 cells. BAG-4 tagged with green fluorescent protein (GFP) or FLAG was localized both in cytoplasm of mouse BNLCL.2 liver cells and human liver hepatoma HepG2 cells. After heat shock, GFP-BAG-4 co-localizes with Hsc70 in the nucleus in HepG2 cells, whereas GFP-BAG-4 mutants lacking the BAG domain remain perinuclear. BAG-4 has no effects on apoB mRNA editing in vitro. However, unlike other apobec-1 complementation factors studied to date, antisense knockdown of BAG-4 in BNLCL.2 cells and in MCR-RH7777 cells increases rather than decreases endogenous apoB mRNA editing. Overexpression of BAG-4 in MCR-RH7777 cells also suppresses apoB mRNA editing. ApoB-48 production also increases with antisense BAG-4 expression in MCR-RH7777 cells. We previously showed that apoB mRNA editing is an intranuclear event (Lau, P. P., Xiong, W. J., Zhu, H. J., Chen, S. H., and Chan, L. (1991) J. Biol. Chem. 266, 20550-20554). Thus, BAG-4 overexpression down regulates apoB mRNA editing by shuttling apobec-1 from the intranuclear perinucleolar compartment to the cytoplasm. We propose that BAG-4 functions as a negative regulator for apobec-1-mediated apoB mRNA editing through its ability to suppress the Hsp/Hsc70 chaperone activity and thereby editosome formation and, as a consequence, prevents nuclear localization of the apobec-1 editosome. PMID- 14559897 TI - Pam and its ortholog highwire interact with and may negatively regulate the TSC1.TSC2 complex. AB - Tuberous Sclerosis Complex (TSC) is an autosomal dominant disorder associated with mutations in TSC1, which codes for hamartin, or TSC2, which codes for tuberin. The brain is one of the most severely affected organs, and CNS lesions include cortical tubers and subependymal giant cell astrocytomas, resulting in mental retardation and seizures. Tuberin and hamartin function together as a complex in mammals and Drosophila. We report here the association of Pam, a protein identified as an interactor of Myc, with the tuberin-hamartin complex in the brain. The C terminus of Pam containing the RING zinc finger motif binds to tuberin. Pam is expressed in embryonic and adult brain as well as in cultured neurons. Pam has two forms in the rat CNS, an approximately 450-kDa form expressed in early embryonic stages and an approximately 350-kDa form observed in the postnatal period. In cortical neurons, Pam co-localizes with tuberin and hamartin in neurites and growth cones. Although Pam function(s) are yet to be defined, the highly conserved Pam homologs, HIW (Drosophila) and RPM-1 (Caenorhabditis elegans), are neuron-specific proteins that regulate synaptic growth. Here we show that HIW can genetically interact with the Tsc1.Tsc2 complex in Drosophila and could negatively regulate Tsc1.Tsc2 activity. Based on genetic studies, HIW has been implicated in ubiquitination, possibly functioning as an E3 ubiquitin ligase through the RING zinc finger domain. Therefore, we hypothesize that Pam, through its interaction with tuberin, could regulate the ubiquitination and proteasomal degradation of the tuberin-hamartin complex particularly in the CNS. PMID- 14559898 TI - Isolation and functional characterization of Ca2+/H+ antiporters from cyanobacteria. AB - Genome sequences of cyanobacteria, Synechocystis sp. PCC 6803, Anabaena sp. PCC 7120, and Thermosynechococcus elongatus BP-1 revealed the presence of a single Ca2+/H+ antiporter in these organisms. Here, we isolated the putative Ca2+/H+ antiporter gene from Synechocystis sp. PCC 6803 (synCAX) as well as a homologous gene from a halotolerant cyanobacterium Aphanothece halophytica (apCAX). In contrast to plant vacuolar CAXs, the full-length apCAX and synCAX genes complemented the Ca2+-sensitive phenotype of an Escherichia coli mutant. ApCAX and SynCAX proteins catalyzed specifically the Ca2+/H+ exchange reaction at alkaline pH. Immunological analysis suggested their localization in plasma membranes. The Synechocystis sp. PCC 6803 cells disrupted of synCAX exhibited lower Ca2+ efflux activity and a salt-sensitive phenotype. Overexpression of ApCAX and SynCAX enhanced the salt tolerance of Synechococcus sp. PCC 7942 cells. Mutagenesis analyses indicate the importance of two conserved acidic amino acid residues, Glu-74 and Glu-324, in the transmembrane segments for the exchange activity. These results clearly indicate that cyanobacteria contain a Ca2+/H+ antiporter in their plasma membranes, which plays an important role for salt tolerance. PMID- 14559900 TI - Bi-phasic effect of interferon (IFN)-alpha: IFN-alpha up- and down-regulates interleukin-4 signaling in human T cells. AB - Interferon (IFN)-alpha/beta is produced by virally infected cells and is believed to play an important role in early phases of the innate immune response. In addition, IFN-alpha/beta inhibits interleukin (IL)-4 signaling in B cells and monocytes, suggesting that IFN-alpha/beta (like IFN-gamma) is a Th1 cytokine. Here, we study cross-talk between IFN-alpha and IL-4 in human T cells. As expected, stimulation with IFN-alpha for 12-24 h inhibits IL-4 signaling. Surprisingly, however, IFN-alpha has the opposite effect on IL-4 signaling at earlier time points (up to 6 h). Thus, IFN-alpha enhances IL-4-mediated STAT6 activation in both CD4+ and CD8+ human T cells. The effect is specific because (i) another interferon, IFN-gamma, does not enhance IL-4-mediated STAT6 activation, (ii) IFN-alpha-mediated STAT1 and STAT2 activation is not modulated by IL-4, and (iii) activation of Janus kinases is not enhanced or prolonged by simultaneous stimulation with IFN-alpha and IL-4. Moreover, co-stimulation results in a selective increased STAT6/STAT2 association and an association between IFNAR/IL-4R components, suggesting that the IFNAR provides an additional STAT6 docking site via STAT2, leading to a more efficient dimerization/activation of STAT6 only. The co-stimulatory effect on STAT6 activation correlates with a cooperative increase in nuclear translocation, DNA binding, transcriptional activity, and mRNA expression of STAT6 target genes (IL-4Ralpha and IL-15Ralpha). In conclusion, we provide evidence that IFN-alpha both up- and down-regulates IL 4-mediated STAT6 signaling and thereby regulates the sensitivity to IL-4 in human T lymphocytes. Thus, our findings suggest that IFN-alpha has a complex regulatory role in adaptive immunity, which is different from the "classical" Th1 profile of IFN-gamma. PMID- 14559899 TI - Pleiotropic effects of Ubp6 loss on drug sensitivities and yeast prion are due to depletion of the free ubiquitin pool. AB - Mutation of the mouse Usp14 gene, encoding the homolog of yeast deubiquitinating enzyme Ubp6, causes ataxia. Here we show that deletion of the UBP6 gene in Saccharomyces cerevisiae causes sensitivity to a broad range of toxic compounds and antagonizes phenotypic expression and de novo induction of the yeast prion [PSI+], a functionally defective self-perpetuating isoform of the translation termination factor Sup35. Conversely, overexpression of ubiquitin (Ub) increases phenotypic expression and induction of [PSI+] in the wild type cells and suppresses all tested ubp6Delta defects, indicating that they are primarily due to depletion of cellular Ub levels. Several lines of evidence suggest that Ubp6 functions on the proteasome. First, Ub levels in the ubp6Delta cells can be partly restored by proteasome inhibitors, suggesting that deletion of Ubp6 decreases Ub levels by increasing proteasome-dependent degradation of Ub. Second, fluorescence microscopy analysis shows that Ubp6-GFP fusion protein is localized to the nucleus of yeast cell, as are most proteasomes. Third, the N-terminal Ub like domain, although it is not required for nuclear localization of Ubp6, targets Ubp6 to the proteasome and cannot be functionally replaced by Ub. The human ortholog of Ubp6, USP14, probably plays a similar role in higher eukaryotes, since it fully compensates for ubp6Delta defects and binds to the yeast proteasome. These data link the Ub system to prion expression and propagation and have broad implications for other neuronal inclusion body diseases. PMID- 14559901 TI - Two regions of cadherin cytoplasmic domains are involved in suppressing motility of a mammary carcinoma cell line. AB - E-cadherin has been termed an "invasion suppressor," yet the mechanism of this suppression is not known. In contrast, several reports indicate N-cadherin does not suppress but, rather, promotes cell motility and invasion. Here, by characterizing a series of chimeric cadherins we defined a previously uncharacterized region consisting of the transmembrane domain and an adjacent portion of the cytoplasmic segment that is responsible for the difference in ability of E- and N-cadherin to suppress movement of mammary carcinoma cells, as quantified from time-lapse video recordings. A mutation in this region enabled N cadherin to suppress motility, indicating that both E- and N-cadherin can suppress, but the activity of N-cadherin is latent, presumably repressed by binding of a specific inhibitor. To define regions common to E- and N-cadherin that are required for suppression, we analyzed a series of deletion mutants. We found that suppression of movement requires E-cadherin amino acids 699-710. Strikingly, beta-catenin binding is not sufficient for and p120ctn is not involved in suppression of these mammary carcinoma cells. Furthermore, the comparable region of N-cadherin can substitute for this required region in E cadherin and is required for suppression by the mutant form of N-cadherin that is capable of suppressing. Variations in expression of factors that bind to the two regions we have identified may explain previously observed differences in response of tumor cells to cadherins. PMID- 14559903 TI - Isolation of temperature-sensitive p53 mutations from a comprehensive missense mutation library. AB - Temperature-sensitive (ts) mutations have been used as a genetic and molecular tool to study the functions of many gene products. Each ts mutant protein may contain a temperature-dependent intramolecular mechanism such as ts conformational change. To identify key ts structural elements controlling the protein function, we screened ts p53 mutants from a comprehensive mutation library consisting of 2,314 p53 missense mutations for their sequence-specific transactivity through p53-binding sequences in Saccharomyces cerevisiae. We isolated 142 ts p53 mutants, including 131 unreported ts mutants. These mutants clustered in beta-strands in the DNA-binding domain, particularly in one of the two beta-sheets of the protein, and 15 residues (Thr155, Arg158, Met160, Ala161, Val172, His214, Ser215, Pro223, Thr231, Thr253, Ile254, Thr256, Ser269, Glu271, and Glu285) were ts hot spots. Among the 142 mutants, 54 were examined further in human osteosarcoma Saos-2 cells, and it was confirmed that 89% of the mutants were also ts in mammalian cells. The ts mutants represented distinct ts transactivities for the p53 binding sequences and a distinct epitope expression pattern for conformation-specific anti-p53 antibodies. These results indicated that the intramolecular beta-sheet in the core DNA-binding domain of p53 was a key structural element controlling the protein function and provided a clue for finding a molecular mechanism that enables the rescue of the mutant p53 function. PMID- 14559902 TI - Phospholipid transfer protein interacts with and stabilizes ATP-binding cassette transporter A1 and enhances cholesterol efflux from cells. AB - Phospholipid lipid transfer protein (PLTP) is ubiquitously expressed in animal tissues and plays multiple roles in lipoprotein metabolism, but the function of peripheral PLTP is still poorly understood. Here we show that one of its possible functions is to transport cholesterol and phospholipids from cells to lipoprotein particles by a process involving PLTP interactions with cellular ATP-binding cassette transporter A1 (ABCA1). When ABCA1 was induced in murine macrophages or ABCA1-transfected baby hamster kidney cells, PLTP gained the ability to promote cholesterol and phospholipid efflux from cells. Although PLTP alone had lipid efflux activity, its maximum activity was observed in the presence of high density lipoprotein particles. Pulsechase studies showed that the interaction of PLTP with ABCA1-expressing cells played a role in promoting lipid efflux. Overexpression of ABCA1 dramatically increased binding of both PLTP and apoA-I to common sites on the cell surface. Both PLTP and apoA-I were covalently cross linked to ABCA1, each protein blocked cross-linking of the other, and both PLTP and apoA-I stabilized ABCA1 protein. These results are consistent with PLTP and apoA-I binding to ABCA1 at the same or closely related sites. Thus, PLTP mimics apolipoproteins in removing cellular lipids by the ABCA1 pathway, except that PLTP acts more as an intermediary in the transfer of cellular lipids to lipoprotein particles. PMID- 14559904 TI - Silencing of RNA helicase II/Gualpha inhibits mammalian ribosomal RNA production. AB - The intricate production of ribosomal RNA is well defined in yeast, but its complexity in higher organisms is barely understood. We recently showed that down regulation of nucleolar protein RNA helicase II/Gualpha (RH-II/Gualpha or DDX21) in Xenopus oocytes inhibited processing of 20 S rRNA to 18 S and contributed to degradation of 28 S rRNA (Yang, H., Zhou, J., Ochs, R. L., Henning, D., Jin, R., and Valdez, B. C. (2003) J. Biol. Chem. 278, 38847-38859). Since no nucleolar RNA helicase has been functionally characterized in mammalian cells, we used short interfering RNA to search for functions for RH-II/Gualpha and its paralogue RH II/Gubeta in rRNA production. Silencing of RH-II/Gualpha by more than 80% in HeLa cells resulted in an almost 80% inhibition of 18 and 28 S rRNA production. This inhibition could be reversed by exogenous expression of wild type RH-II/Gualpha. A helicase-deficient mutant form having ATPase activity was able to rescue the production of 28 S but not 18 S rRNA. A phenotype exhibiting inhibition of 18 S and 28 S rRNA production was also observed when the paralogue RH-II/Gubeta was overexpressed. Both down-regulation of RH-II/Gualpha and overexpression of RH II/Gubeta slowed cell proliferation. The opposite effects of the two paralogues suggest antagonistic functions. PMID- 14559905 TI - Sequential binding of agonists to the beta2 adrenoceptor. Kinetic evidence for intermediate conformational states. AB - The beta2 adrenoreceptor (beta2AR) is a prototypical G protein-coupled receptor (GPCR) activated by catecholamines. Agonist activation of GPCRs leads to sequential interactions with heterotrimeric G proteins, which activate cellular signaling cascades, and with GPCR kinases and arrestins, which attenuate GPCR mediated signaling. We used fluorescence spectroscopy to monitor catecholamine induced conformational changes in purified beta2AR. Here we show that upon catecholamine binding, beta2ARs undergo transitions to two kinetically distinguishable conformational states. Using a panel of chemically related catechol derivatives, we identified the specific chemical groups on the agonist responsible for the rapid and slow conformational changes in the receptor. The conformational changes observed in our biophysical assay were correlated with biologic responses in cellular assays. Dopamine, which induces only a rapid conformational change, is efficient at activating Gs but not receptor internalization. In contrast, norepinephrine and epinephrine, which induce both rapid and slow conformational changes, are efficient at activating Gs and receptor internalization. These results support a mechanistic model for GPCR activation where contacts between the receptor and structural determinants of the agonist stabilize a succession of conformational states with distinct cellular functions. PMID- 14559906 TI - Regulation of SPIN90 phosphorylation and interaction with Nck by ERK and cell adhesion. AB - SPIN90 is a widely expressed Nck-binding protein that contains one Src homology 3 (SH3) domain, three Pro-rich motifs, and a serine/threonine-rich region, and is known to participate in sarcomere assembly during cardiac myocyte differentiation. We used in vitro binding assays and yeast two-hybrid screening analysis to identify Nck, betaPIX, Wiscott-Aldrich syndrome protein (WASP), and ERK1 as SPIN90-binding proteins. It appears that betaPIX, WASP, and SPIN90 form a complex that interacts with Nck in a manner dependent upon cell adhesion to extracellular matrix. The betaPIX.WASP.SPIN90.Nck interaction was abolished in suspended and cytochalasin D-treated cells, but was recovered when cells were replated on fibronectin-coated dishes. The SPIN90.betaPIX.WASP complex was stable, even in suspended cells, suggesting SPIN90 serves as an adaptor molecule to recruit other proteins to Nck at focal adhesions. In addition, we found that overexpression of the SPIN90 SH3 domain or Pro-rich region, respectively, abolished SPIN90.Nck and SPIN90.betaPIX interactions, resulting in detachment of cells from extracellular matrix. SPIN90 was phosphorylated by ERK1, which was, itself, activated by cell adhesion and platelet-derived growth factor. Such phosphorylation of SPIN90 likely promotes the interaction of the SPIN90.betaPIX.WASP complex and Nck. It thus appears that the interaction of the betaPIX.WASP.SPIN90 complex with Nck is crucial for stable cell adhesion and can be dynamically modulated by SPIN90 phosphorylation that is dependent on cell adhesion and ERK activation. PMID- 14559907 TI - Unique mechanism of action of the thiourea drug isoxyl on Mycobacterium tuberculosis. AB - The thiourea isoxyl (thiocarlide; 4,4'-diisoamyloxydiphenylthiourea) is known to be an effective anti-tuberculosis drug, active against a range of multidrug resistant strains of Mycobacterium tuberculosis and has been used clinically. Little was known of its mode of action. We now demonstrate that isoxyl results in a dose-dependent decrease in the synthesis of oleic and, consequently, tuberculostearic acid in M. tuberculosis with complete inhibition at 3 microg/ml. Synthesis of mycolic acid was also affected. The anti-bacterial effect of isoxyl was partially reversed by supplementing growth medium with oleic acid. The specificity of this inhibition pointed to a Delta9-stearoyl desaturase as the drug target. Development of a cell-free assay for Delta9-desaturase activity allowed direct demonstration of the inhibition of oleic acid synthesis by isoxyl. Interestingly, sterculic acid, a known inhibitor of Delta9-desaturases, emulated the effect of isoxyl on oleic acid synthesis but did not affect mycolic acid synthesis, demonstrating the lack of a relationship between the two effects of the drug. The three putative fatty acid desaturases in the M. tuberculosis genome, desA1, desA2, and desA3, were cloned and expressed in Mycobacterium bovis BCG. Cell-free assays and whole cell labeling demonstrated increased Delta9 desaturase activity and oleic acid synthesis only in the desA3-overexpressing strain and an increase in the minimal inhibitory concentration for isoxyl, indicating that DesA3 is the target of the drug. These results validate membrane bound Delta9-desaturase, DesA3, as a new therapeutic target, and the thioureas as anti-tuberculosis drugs worthy of further development. PMID- 14559908 TI - Importance of TRF1 for functional telomere structure. AB - Telomeres are comprised of telomeric DNA sequences and associated binding molecules. Their structure functions to protect the ends of linear chromosomes and ensure chromosomal stability. One of the mammalian telomere-binding factors, TRF1, localizes telomeres by binding to double-stranded telomeric DNA arrays. Because the overexpression of wild-type and dominant-negative TRF1 induces progressive telomere shortening and elongation in human cells, respectively, a proposed major role of TRF1 is that of a negative regulator of telomere length. Here we report another crucial function of TRF1 in telomeres. In conditional mouse TRF1 null mutant embryonic stem cells, TRF1 deletion induced growth defect and chromosomal instability. Although no clear telomere shortening or elongation was observed in short term cultured TRF1-deficient cells, abnormal telomere signals were observed, and TRF1-interacting telomere-binding factor, TIN2, lost telomeric association. Furthermore, another double-stranded telomeric DNA-binding factor, TRF2, also showed decreased telomeric association. Importantly, end-to end fusions with detectable telomere signals at fusion points accumulated in TRF1 deficient cells. These results strongly suggest that TRF1 interacts with other telomere-binding molecules and integrates into the functional telomere structure. PMID- 14559909 TI - Reversible oxidation of the active site cysteine of peroxiredoxins to cysteine sulfinic acid. Immunoblot detection with antibodies specific for the hyperoxidized cysteine-containing sequence. AB - We previously suggested that oxidation of the active site cysteine of peroxiredoxin (Prx) I or Prx II to cysteine sulfinic acid in H2O2-treated cells is reversible (Woo, H. A., Chae, H. Z., Hwang, S. C., Yang, K.-S., Kang, S. W., Kim, K., and Rhee, S. G. (2003) Science 300, 653-656). In contrast, it was recently proposed that sulfinylation of Prx II, but not that of Prx I or Prx III, is reversible (Chevallet, M., Wagner, E., Luche, S., van Dorssealaer, A., Leize Wagner, E., and Rabilloud, T. (2003) J. Biol. Chem. 278, 37146-37153). The detection of sulfinylated proteins in both of these previous studies relied on complex proteomics analysis. We now describe a simple immunoblot assay for the detection of sulfinylated Prx enzymes that is based on antibodies produced in response to a sulfonylated peptide modeled on the conserved active site sequence. These antibodies recognized both sulfinic and sulfonic forms of Prx equally well and allowed the detection of sulfinylated Prx enzymes in H2O2-treated cells with high sensitivity and specificity. With the use of these antibodies, we demonstrated that not only the cytosolic enzymes Prx I and Prx II but also the mitochondrial enzyme Prx III undergo reversible sulfinylation. The generation of antibodies specific for sulfonylated peptides should provide insight into protein function similar to that achieved with antibodies to peptides containing phosphoserine or phosphothreonine. PMID- 14559911 TI - Differential recruitment of Kv1.4 and Kv4.2 to lipid rafts by PSD-95. AB - The activity of voltage-gated potassium (Kv) channels, and consequently their influence on cellular functions, can be substantially altered by phosphorylation. Several protein kinases that modulate Kv channel activity are found in membrane subdomains known as lipid rafts, which are thought to organize signaling complexes in the cell. Thus, we asked whether Kv1.4 and Kv4.2, two channels with critical roles in excitable cells, are found in lipid rafts. Acylation can target proteins to raft regions; however, Kv channels are not acylated, and therefore, a different mechanism must exist to bring them into these membrane subdomains. Because both Kv1.4 and Kv4.2 interact with postsynaptic density protein 95 (PSD 95), which is acylated (specifically, palmitoylated), we examined whether PSD-95 can recruit these channels to lipid rafts. We found that a portion of Kv1.4 and Kv4.2 protein in rat brain membranes is raft-associated. Lipid raft patching and immunostaining confirmed that some Kv4.2 is in Thy-1-containing rafts in rat hippocampal neurons. Using a heterologous expression system, we determined that palmitoylation of PSD-95 was crucial to its localization to lipid rafts. We then assessed the contribution of PSD-95 to the raft association of these channels. Co expression of PSD-95 increased the amount of Kv1.4, but not Kv4.2, in lipid rafts. Deleting the PSD-95 binding motif of Kv1.4 eliminated this recruitment, as did substituting a palmitoylation-deficient PSD-95 mutant. This work represents the first evidence that PSD-95 binding can recruit Kv channels into lipid rafts, a process that could facilitate interactions with the protein kinases that affect channel activity. PMID- 14559912 TI - Increased expression of Bcl-xL and c-Myc is associated with transformation by Abelson murine leukemia virus. AB - Transformation mediated by the v-Abl oncoprotein, a tyrosine kinase encoded by the Abelson murine leukemia virus, is a multi-step process requiring genetic alterations in addition to expression of v-Abl. Loss of p53 or p19ARF was previously shown to be required for Abelson murine leukemia virus transformation of primary mouse embryonic fibroblasts (MEFs). By comparing gene expression patterns in primary p53-/- MEFs acutely infected with the v-Abl retrovirus, v-Abl transformed MEF clones, and v-Abl-transformed MEF clones treated with Abl kinase inhibitor STI 571, we have identified additional genetic alterations associated with v-Abl transformation. Bcl-xL mRNA was elevated in three of five v-Abl transformed MEF clones. In addition, elevated expression of c-Myc mRNA, caused either by c-myc gene amplification or by enhanced signaling via STAT3, was observed in five v-Abl-transformed MEF clones. The data suggest that increases in cell survival associated with Bcl-xL and increases in cell growth associated with c-Myc facilitate the transformation process dependent on constitutive mitogenic signaling by v-Abl. PMID- 14559910 TI - The voltage-dependent calcium channel beta subunit contains two stable interacting domains. AB - Voltage-dependent calcium channels selectively enable Ca2+ ion movement through cellular membranes. These multiprotein complexes are involved in a wide spectrum of biological processes such as signal transduction and cellular homeostasis. alpha1 is the membrane pore-forming subunit, whereas beta is an intracellular subunit that binds to alpha1, facilitating and modulating channel function. We have expressed, purified, and characterized recombinant beta3 and beta2a using both biochemical and biophysical methods, including electrophysiology, to better understand the beta family's protein structural and functional correlates. Our results indicate that the beta protein is composed of two distinct domains that associate with one another in a stable manner. The data also suggest that the polypeptide regions outside these domains are not structured when beta is not in complex with the channel. In addition, the beta structural core, comprised of just these two domains without other sequences, binds tightly to the alpha interaction domain (AID) motif, a sequence derived from the alpha1 subunit and the principal anchor site of beta. Domain II is responsible for this binding, but domain I enhances it. PMID- 14559913 TI - The contribution of amino acid region ASP695-TYR698 of factor V to procofactor activation and factor Va function. AB - There is strong evidence that a functionally important cluster of amino acids is located on the COOH-terminal portion of the heavy chain of factor Va, between amino acid residues 680 and 709. To ascertain the importance of this region for cofactor activity, we have synthesized five overlapping peptides representing this amino acid stretch (10 amino acids each, HC1-HC5) and tested them for inhibition of prothrombinase assembly and function. Two peptides, HC3 (spanning amino acid region 690-699) and HC4 (containing amino acid residues 695-704), were found to be potent inhibitors of prothrombinase activity with IC(50) values of approximately 12 and approximately 10 microm, respectively. The two peptides were unable to interfere with the binding of factor Va to active site fluorescently labeled Glu-Gly-Arg human factor Xa, and kinetic analyses showed that HC3 and HC4 are competitive inhibitors of prothrombinase with respect to prothrombin with K(i) values of approximately 6.3 and approximately 5.3 microm, respectively. These data suggest that the peptides inhibit prothrombinase because they interfere with the incorporation of prothrombin into prothrombinase. The shared amino acid motif between HC3 and HC4 is composed of Asp(695)-Tyr-Asp-Tyr-Gln(699) (DYDYQ). A pentapeptide with this sequence inhibited both prothrombinase function with an IC(50) of 1.6 microm (with a K(D) for prothrombin of 850 nm), and activation of factor V by thrombin. Peptides HC3, HC4, and DYDYQ were also found to interact with immobilized thrombin. A recombinant factor V molecule with the mutations Asp(695) --> Lys, Tyr(696) --> Phe, Asp(697) --> Lys, and Tyr(698) --> Phe (factor V(2K2F)) was partially resistant to activation by thrombin but could be readily activated by RVV-V activator (factor Va(RVV)(2K2F)) and factor Xa (factor Va(Xa)(2K2F)). Factor Va(RVV)(2K2F) and factor Va(Xa)(2K2F) had impaired cofactor activity within prothrombinase in a system using purified reagents. Our data demonstrate for the first time that amino acid sequence 695-698 of factor Va heavy chain is important for procofactor activation and is required for optimum prothrombinase function. These data provide functional evidence for an essential and productive contribution of factor Va to the activity of prothrombinase. PMID- 14559914 TI - Cytoplasmic p21Cip1 is involved in Ras-induced inhibition of the ROCK/LIMK/cofilin pathway. AB - Accumulating evidence suggests that p21(Cip1) located in the cytoplasm might play a role in promoting transformation and tumor progression. Here we show that oncogenic H-RasV12 contributes to the loss of actin stress fibers by inducing cytoplasmic localization of p21(Cip1), which uncouples Rho-GTP from stress fiber formation by inhibiting Rho kinase (ROCK). Concomitant with the loss of stress fibers in Ras-transformed cells, there is a decrease in the phosphorylation level of cofilin, which is indicative of a compromised ROCK/LIMK/cofilin pathway. Inhibition of MEK in Ras-transformed NIH3T3 results in restoration of actin stress fibers accompanied by a loss of cytoplasmic p21(Cip1), and increased phosphorylation of cofilin. Ectopic expression of cytoplasmic but not nuclear p21(Cip1) in Ras-transformed cells was effective in preventing stress fibers from being restored upon MEK inhibition and inhibited phosphorylation of cofilin. p21(Cip1) was also found to form a complex with ROCK in Ras-transformed cells in vivo. Furthermore, inhibition of the PI 3-kinase pathway resulted in loss of p21(Cip1) expression accompanied by restoration of phosphocofilin, which was not accompanied by stress fiber formation. These results suggest that restoration of cofilin phosphorylation in Ras-transformed cells is necessary but not sufficient for stress fiber formation. Our findings define a novel mechanism for coupling cytoplasmic p21(Cip1) to the control of actin polymerization by compromising the Rho/ROCK/LIMK/cofilin pathway by oncogenic Ras. These studies suggest that localization of p21(Cip1) to the cytoplasm in transformed cells contributes to pathways that favor not only cell proliferation, but also cell motility thereby contributing to invasion and metastasis. PMID- 14559915 TI - Identification of a secondary zinc-binding site in staphylococcal enterotoxin C2. Implications for superantigen recognition. AB - The previously determined crystal structure of the superantigen staphylococcal enterotoxin C2 (SEC2) showed binding of a single zinc ion located between the N- and C-terminal domains. Here we present the crystal structure of SEC2 determined to 2.0 A resolution in the presence of additional zinc. The structure revealed the presence of a secondary zinc-binding site close to the major histocompatibility complex (MHC)-binding site of the toxin and some 28 A away from the primary zinc-binding site of the toxin found in previous studies. T cell stimulation assays showed that varying the concentration of zinc ions present affected the activity of the toxin and we observed that high zinc concentrations considerably inhibited T cell responses. This indicates that SEC2 may have multiple modes of interaction with the immune system that are dependent on serum zinc levels. The potential role of the secondary zinc-binding site and that of the primary one in the formation of the TCR.SEC2.MHC complex are considered, and the possibility that zinc may regulate the activity of SEC2 as a toxin facilitating different T cell responses is discussed. PMID- 14559916 TI - Genetic complementation in yeast reveals functional similarities between the catalytic subunits of mammalian signal peptidase complex. AB - Type I signal peptidases (SPs) comprise a family of structurally related enzymes that cleave signal peptides from precursor proteins following their transport out of the cytoplasmic space in eukaryotic and prokaryotic cells. One such enzyme, the mitochondrial inner membrane peptidase, has two catalytic subunits, which recognize distinct cleavage site motifs in their signal peptide substrates. The only other known type I SP with two catalytic subunits is the signal peptidase complex (SPC) in the mammalian endoplasmic reticulum. Here, we tested the hypothesis that, as with inner membrane peptidase catalytic subunits, SPC catalytic subunits exhibit nonoverlapping substrate specificity. We constructed two yeast strains without endogenous SP, one expressing canine SPC18 and the other expressing a truncation of canine SPC21 (SPC21 Delta N), which lacks 24 N terminal residues that prevent expression of SPC21 in yeast. By monitoring a variety of soluble and membrane-bound substrates, we find that, in contrast to the tested hypothesis, SPC catalytic subunits exhibit overlapping substrate specificity. SPC18 and SPC21 Delta N do, however, cleave some substrates with different efficiencies, although no pattern for this behavior could be discerned. In light of the functional similarities between SPC proteins, we developed a membrane protein fragmentation assay to monitor the position of the catalytic sites relative to the surface of the endoplasmic reticulum membrane. Using this assay, our results suggest that the active sites of SPC18 and SPC21 Delta N are located 4-11 A above the membrane surface. These data, thus, support a model that SPC18 and SPC21 are functionally and structurally similar to each other. PMID- 14559917 TI - The SK3 subunit of small conductance Ca2+-activated K+ channels interacts with both SK1 and SK2 subunits in a heterologous expression system. AB - The aim of this study was to determine whether functional heteromeric channels can be formed by co-assembly of rat SK3 (rSK3) potassium channel subunits with either SK1 or SK2 subunits. First, to determine whether rSK3 could co-assemble with rSK2 we created rSK3VK (an SK3 mutant insensitive to block by UCL 1848). When rSK3VK was co-expressed with rSK2 the resulting currents had an intermediate sensitivity to UCL 1848 (IC50 of approximately 5 nM compared with 120 pM for rSK2 and >300 nM for rSK3VK), suggesting that rSK3 and rSK2 can form functional heteromeric channels. To detect co-assembly of SK3 with SK1, we initially used a dominant negative construct of the human SK1 subunit (hSK1YP). hSK1YP dramatically reduced the SK3 current, supporting the idea that SK3 and SK1 subunits also interact. To determine whether these assemblies were functional we created rSK3VF, an rSK3 mutant with an enhanced affinity for tetraethylammonium chloride (TEA) (IC50 of 0.3 mM). Co-transfection of rSK3VF and hSK1 produced currents with a sensitivity to TEA not different from that of hSK1 alone (IC50 approximately 15 mM). These results suggest that hSK1 does not produce functional cell-surface assemblies with SK3. Antibody-staining experiments suggested that hSK1 may reduce the number of functional SK3 subunits reaching the cell surface. Additional experiments showed that co-expression of the rat SK1 gene with SK3 also dramatically suppressed SK current. The pharmacology of the residual current was consistent with that of homomeric SK3 assemblies. These results demonstrate interactions that cause changes in protein trafficking, cell surface expression, and channel pharmacology and strongly suggest heteromeric assembly of SK3 with the other SK channel subunits. PMID- 14559918 TI - PIASgamma represses the transcriptional activation induced by the nuclear receptor Nurr1. AB - Nurr1 is a transcription factor essential for the development of ventral dopaminergic neurons. In search for regulatory mechanisms of Nurr1 function, we identified the SUMO (small ubiquitin-like modifier)-E3 ubiquitin-protein isopeptide ligase, PIASgamma, as an interaction partner of Nurr1. Overexpressed PIASgamma and Nurr1 co-localize in the nuclei of transfected cells, and their interaction is demonstrated through co-immunoprecipitation and glutathione S transferase pulldown assays. Co-expression of PIASgamma with Nurr1 results in a potent repression of Nurr1-dependent transcriptional activation of an artificial NGFI-B response element (NBRE) reporter as well as of a reporter driven by the native tyrosine hydroxylase promoter. We identified two consensus sumoylation sites in Nurr1. The substitution of lysine 91 by arginine in one SUMO site enhanced the transcriptional activity of Nurr1, whereas the substitution of lysine 577 by arginine in the second SUMO site decreased transcriptional activity of Nurr1. Interestingly, PIASgamma-induced repression of Nurr1 activity does not require the two sumoylation sites, because each mutant is repressed as efficiently as the wild type Nurr1. In addition, the mutations do not alter Nurr1 nuclear localization. Finally, we provide evidence that Nurr1 and PIASgamma co exist in several nuclei of the rodent central nervous system by demonstrating the co-expression of Nurr1 protein and PIASgamma mRNA in the same cells. In conclusion, our studies identified PIASgamma as a transcriptional co-regulator of Nurr1 and suggest that this interaction may have a physiological role in regulating the expression of Nurr1 target genes. PMID- 14559919 TI - The alpha2 isoform of Na,K-ATPase mediates ouabain-induced cardiac inotropy in mice. AB - Inhibition of Na,K-ATPase activity by cardiac glycosides is believed to be the major mechanism by which this class of drugs increases heart contractility. However, direct evidence demonstrating this is lacking. Furthermore it is unknown which specific alpha isoform of Na,K-ATPase is responsible for the effect of cardiac glycosides. Several studies also suggest that cardiac glycosides, such as ouabain, function by mechanisms other than inhibition of the Na,K-ATPase. To determine whether Na,K-ATPase, specifically the alpha2 Na,K-ATPase isozyme, mediates ouabain-induced cardiac inotropy, we developed animals expressing a ouabain-insensitive alpha2 isoform of the Na,K-ATPase using Cre-Lox technology and analyzed cardiac contractility after administration of ouabain. The homozygous knock-in animals were born in normal Mendelian ratio and developed normally to adulthood. Analysis of their cardiovascular function demonstrated normal heart function. Cardiac contractility analysis in isolated hearts and in intact animals demonstrated that ouabain-induced cardiac inotropy occurred in hearts from wild type but not from the targeted animals. These results clearly demonstrate that the Na,K-ATPase and specifically the alpha2 Na,K-ATPase isozyme mediates ouabain-induced cardiac contractility in mice. PMID- 14559920 TI - AKT/protein kinase B regulation of BCL family members during oxysterol-induced apoptosis. AB - Cells of the vasculature, including macrophages, smooth muscle cells, and endothelial cells, exhibit apoptosis in culture upon treatment with oxidized low density lipoprotein, as do vascular cells of atherosclerotic plaque. Several lines of evidence support the hypothesis that the apoptotic component of oxidized low density lipoprotein is one or more oxysterols, which have been shown to induce apoptosis through the mitochondrial pathway. Activation of the mitochondrial pathway of apoptosis is regulated by members of the BCL family of proteins. In this study, we demonstrate that, in the murine macrophage-like cell line P388D1, oxysterols (25-hydroxycholesterol and 7-ketocholesterol) induced the degradation of the prosurvival protein kinase AKT (protein kinase B). This led, in turn, to the activation of the BCL-2 homology-3 domain-only proteins BIM and BAD and down-regulation of the anti-apoptotic multi-BCL homology domain protein BCL-xL. These responses would be expected to activate the pro-apoptotic multi-BCL homology domain proteins BAX and BAK, leading to the previously reported release of cytochrome c observed during oxysterol-induced apoptosis. Somewhat surprisingly, small interfering RNA knockdown of BAX resulted in a complete block of the induction of apoptosis by 25-hydroxycholesterol. PMID- 14559921 TI - A context-dependent ClpX recognition determinant located at the C terminus of phage Mu repressor. AB - The bacteriophage Mu immunity repressor is a conformationally sensitive sensor that can be interconverted between forms resistant to and sensitive to degradation by ClpXP protease. Protease-sensitive repressor molecules with an altered C-terminal sequence promote rapid degradation of the wild-type repressor by inducing its C-terminal end to become exposed. Here we determined that the last 5 C-terminal residues (CTD5) of the wild-type repressor contain the motif required for recognition by the ClpX molecular chaperone, a motif that is strongly dependent upon the context in which it is presented. Although attachment of the 11-residue ssrA degradation tag to the C terminus of green fluorescent protein (GFP) promoted its rapid degradation by ClpXP, attachment of 5-27 C terminal residues of the repressor failed to promote degradation. Disordered peptides derived from 41 and 35 C-terminal residues of CcdA (CcdA41) and thioredoxin (TrxA35), respectively, activated CTD5 when placed as linkers between GFP and repressor C-terminal sequences. However, when the entire thioredoxin sequence was included as a linker to promote an ordered configuration of the TrxA35 peptide, the resulting substrate was not degraded. In addition, a hybrid tag, in which CTD5 replaced the 3-residue recognition motif of the ssrA tag, was inactive when attached directly to GFP but active when attached through the CcdA41 peptide. Thus, CTD5 is sufficient to act as a recognition motif but has requirements for its presentation not shared by the ssrA tag. We suggest that activation of CTD5 may require presentation on a disordered or flexible domain that confers ligand flexibility. PMID- 14559922 TI - The fidelity of HPV16 E1/E2-mediated DNA replication. AB - Human papillomaviruses (HPV) are causative agents in a variety of human diseases; for example over 99% of cervical carcinomas contain HPV DNA sequences. Often in cervical carcinoma the HPV genome is integrated into the host genome resulting in unregulated expression of the viral transforming proteins E6 and E7. Therefore viral integration is a step toward HPV-induced carcinogenesis. Integration of the HPV genome could occur following double-strand DNA breaks that could arise during viral DNA replication. We investigated the fidelity of HPV 16 E1- and E2-mediated DNA replication of non-damaged and UVC-damaged templates in a variety of cell lines with different genetic backgrounds; C33a (derived from an HPV-negative cervical carcinoma), XP30RO (deficient in the by-pass polymerase eta (poleta)), XP30eta (expressing a restored wild-type poleta), XP12RO (nucleotide excision repair defective), and MRC5 (derived from a 14-week-old human fetus). The results demonstrate that the fidelity of E1- and E2-mediated DNA replication is reflective of the genetic background in which the assays are carried out. For example, restoring poleta to the XP30 cell line results in a 3-fold drop in the number of mutants obtained following replication of a UVC-damaged template. A relatively high percentage of the mutant-replicated molecules arise as a result of genetic rearrangement. This is the first time such studies have been carried out with an HPV replication system, and the results are discussed in the context of the HPV life cycle and what is known about HPV genomes in human cancers. PMID- 14559923 TI - An essential role in molting and morphogenesis of Caenorhabditis elegans for ACN 1, a novel member of the angiotensin-converting enzyme family that lacks a metallopeptidase active site. AB - Genome sequence analyses predict many proteins that are structurally related to proteases but lack catalytic residues, thus making functional assignment difficult. We show that one of these proteins (ACN-1), a unique multi-domain angiotensin-converting enzyme (ACE)-like protein from Caenorhabditis elegans, is essential for larval development and adult morphogenesis. Green fluorescent protein-tagged ACN-1 is expressed in hypodermal cells, the developing vulva, and the ray papillae of the male tail. The hypodermal expression of acn-1 appears to be controlled by nhr-23 and nhr-25, two nuclear hormone receptors known to regulate molting in C. elegans. acn-1(RNAi) causes arrest of larval development because of a molting defect, a protruding vulva in adult hermaphrodites, severely disrupted alae, and an incomplete seam syncytium. Adult males also have multiple tail defects. The failure of the larval seam cells to undergo normal cell fusion is the likely reason for the severe disruption of the adult alae. We propose that alteration of the ancestral ACE during evolution, by loss of the metallopeptidase active site and the addition of new protein modules, has provided opportunities for novel molecular interactions important for post-embryonic development in nematodes. PMID- 14559924 TI - Mutation of CpGs in the murine stem cell virus retroviral vector long terminal repeat represses silencing in embryonic stem cells. AB - Although DNA methylation and transcriptional repression are generally associated, a causal role for DNA methylation in silencing of retroviral vectors has not been established. The newer generation murine stem cell virus retroviral vector (MSCV) lacks many of the repressive cis-acting DNA sequences identified in Moloney murine leukemia virus but remains sensitive to transcriptional silencing in various cell types. To determine the contribution of cytosine methylation to MSCV silencing, we mutated CpG dinucleotides located in the MSCV long terminal repeat (LTR) that are clustered in the U3 region and directly spanning the transcription start site in the R region. Effects of the CpG mutations on MSCV silencing were assessed in murine embryonic stem cells. An analysis of numerous clonal proviral integrants showed that mutation of CpGs in both clusters eliminated proviral integrants that were completely silenced. Variegated expression was shown to represent a substantial component of intraclonal silencing and was independent of the presence of CpGs in the LTR. Treatment of transduced cells with 5 azadeoxycytidine delayed establishment of the silenced state but had only a modest effect on expression of some proviral integrants at late times post transduction. These results are direct evidence for a causal contribution of DNA methylation in the LTR to MSCV silencing and define the promoter region CpGs as a repressive element in embryonic stem cells. Furthermore, distinct mechanisms are suggested for establishment and maintenance of the silenced proviral state. PMID- 14559925 TI - Importance of weight management in type 2 diabetes: review with meta-analysis of clinical studies. AB - Obesity is a major risk factor for development of diabetes, and excessive energy intake is a major contributor to poor glycemic control in Type 2 diabetes. The impact of obesity on risk for diabetes as well as coronary heart disease (CHD) risk factors and the benefits of weight loss in decreasing risk for developing diabetes and improving glycemia and CHD risks were reviewed. A systematic review of the medical literature to assess the impact of obesity and weight gain on risk for diabetes and CHD was done. We performed a meta-analysis of the effects of weight loss for obese diabetic individuals. Controlled clinical trials assessing lifestyle changes on risk for developing diabetes and weight loss effects on glycemia and CHD risk factors were reviewed. Obesity and weight gain can increase risk for diabetes by greater than ninetyfold and CHD by about sixfold. Very-low energy diets (VLED) decrease fasting plasma glucose values by approximately 50% within two weeks and these changes are sustained with continued energy restriction. Twelve weeks of energy-restricted diets were associated with these significant decreases: body weight, 9.6%; fasting plasma glucose, 25.7%; serum cholesterol, 9.2%; serum triglycerides, 26.7%; systolic blood pressure, 8.1%; and diastolic blood pressure, 8.6%. Larger weight losses were associated with larger reductions in these values. The reviewed data suggest that US health care providers should endorse the American Heart Association's and European diabetes associations' recommendations that diabetic persons achieve and maintain a BMI of or=85th percentile, adjusted for age and gender). Subjects were maintained on a hypocaloric diet with a normal daily mineral content in both treatment groups and received oral treatment with orlistat 120 mg (n = 16) or placebo (n = 16) three times daily for 21 days. Following a 14-day equilibration period, balances for calcium, phosphorus, magnesium, iron, copper and zinc were measured for days 15-21. Serum and urine electrolytes were also measured at baseline and at the end of treatment. RESULTS: On average, orlistat inhibited dietary fat absorption by approximately 27%. This degree of dietary fat inhibition caused no significant changes in mineral balance between orlistat and placebo groups. In addition, serum and urine electrolytes (sodium and potassium) as well as urinary creatinine excretion were not affected by orlistat treatment. Orlistat was well tolerated; adverse events occurred mainly in the gastrointestinal tract and were of mild or moderate intensities. CONCLUSIONS: Administration of orlistat had no significant effect on the balance of six selected minerals in adolescent obese patients. PMID- 14559928 TI - Ingestion of a dietary supplement containing dehydroepiandrosterone (DHEA) and androstenedione has minimal effect on immune function in middle-aged men. AB - OBJECTIVE: This study investigated the effects of four weeks of intake of a supplement containing dehydroepiandrosterone (DHEA), androstenedione and herbal extracts on immune function in middle-aged men. DESIGN: Subjects consumed either an oral placebo or an oral supplement for four weeks. The supplement contained a total daily dose of 150 mg DHEA, 300 mg androstenedione, 750 mg Tribulus terrestris, 625 mg chrysin, 300 mg indole-3-carbinol and 540 mg saw palmetto. MEASUREMENTS: Peripheral blood mononuclear cells were used to assess phytohemagglutinin(PHA)-induced lymphocyte proliferation and cytokine production. The cytokines measured were interleukin (IL)-2, IL-4, IL-10, IL-1beta, and interferon (IFN)-gamma. Serum free testosterone, androstenedione, estradiol, dihydrotestosterone (DHT) were also measured. RESULTS: The supplement significantly increased serum levels of androstenedione, free testosterone, estradiol and DHT during week 1 to week 4. Supplement intake did not affect LPS or ConA proliferation and had minimal effect on PHA-induced proliferation. LPS induced production of IL-1beta, and PHA-induced IL-2, IL-4, IL-10, or IFN-gamma production was not altered by the supplement. The addition of the same supplement, DHEA or androstenedione alone to lymphocyte cultures in vitro did not alter lymphocyte proliferation, IL-2, IL-10, or IFN-gamma, but did increase IL-4. In addition, serum HDL-C concentration significantly declined. CONCLUSION: These findings suggest that, although chronic intake of a complex dietary supplement containing DHEA, androstenedione and herbal extracts increases serum androgen levels, it has minimal effect on immune function in middle-aged men. PMID- 14559929 TI - Vitamin C intake and risk of ischemic heart disease in a population with a high prevalence of smoking. AB - OBJECTIVE: Epidemiological data on the relationship between vitamin C intake and ischemic heart disease (IHD) risk are limited in the Asian population, with a high prevalence of smoking. This study aims to investigate the association between vitamin C intake and the incidence of non-fatal IHD in Korean men. METHODS: The case group consisted of 108 patients with electrocardiogram confirmed myocardial infarction or angiographically confirmed (>or=50% stenosis) coronary artery disease (CAD) who were admitted to a university teaching hospital in Seoul, Korea. The controls were 142 age-matched patients admitted to the departments of ophthalmology and orthopedic surgery at the same hospital. Vitamin C intake was assessed by a nutritionist using a semi-quantitative food frequency method, and body mass index (BMI), tobacco use and past history of cardiovascular disease were determined by examination and interview. RESULTS: After controlling for cardiovascular risk factors, including BMI, smoking, past history of hypertension, past history of hyperlipidemia, dietary intakes of energy, total fat (or subtype of fat), cholesterol, beta-carotene, and vitamin E, the odds ratio (OR) of non-fatal IHD was 0.34 (95% confidence interval (CI) 0.13-0.90) in the highest tertile of vitamin C intake compared with those in the lowest tertile. In a subgroup analysis, which compared nonsmokers in the highest tertile of vitamin C intake to current smokers in the lowest tertile of vitamin C intake, the odds ratio of developing non-fatal IHD was 0.12 (95% CI 0.02-0.77). CONCLUSION: This study suggests that higher intake of vitamin C is associated with the decreased risk of non-fatal IHD in a population with a high prevalence of smoking. PMID- 14559930 TI - Dysprosium chloride as a nonabsorbable gastrointestinal marker for studies of stable isotope-labeled triglyceride excretion in man. AB - OBJECTIVE: The aim of this work was to determine if dysprosium chloride (DyCl(3)) is a suitable nonabsorbable marker for studies of labeled-triglyceride excretion in cystic fibrosis patients allowing excretion to be determined accurately after analysis of one or two stools. METHODS: A series of 66 absorption studies were conducted in 36 young cystic fibrosis patients over a five year period. All tests consisted of ingesting a single test meal containing both (13)C-labeled triglyceride (TG*) and DyCl(3); in most studies the food colorant brilliant blue (FD&C blue #1) was administered along with the DyCl(3). Ingestion of the test meal was followed by collection of individual stools for 72 to 96 hours. Stools were analyzed for (13)C-Excess ((13)C*) and Dy. RESULTS: Excretion of Dy in cystic fibrosis patients who exhibited a wide-range of steatorrhea was quantitative. Fractional excretion of Dy and (13)C* in individual stools showed a high linear correlation (r(2) = 0.969) with a slope and y-intercept close to unity and zero, respectively. As a result, estimates of TG* excretion based on analysis of only two stools (partial pool method, PPM) were not different from those based on the analysis of all stools or stool composites. This was true both when Dy content and when stool color due to ingested brilliant blue was used to determine which stools to analyze for the PPM. CONCLUSIONS: Combining the use of Dy and brilliant blue permits reasonably accurate estimates of fecal TG* excretion after analysis of samples from two easily identified stools. This practical method can be used to address many important clinical and experimental questions regarding triglyceride digestion and absorption that may otherwise go unanswered. PMID- 14559931 TI - Effect of dietary n-3 and n-6 oils with and without food restriction on activity of antioxidant enzymes and lipid peroxidation in livers of cyclophosphamide treated autoimmune-prone NZB/W female mice. AB - OBJECTIVE: Cyclophosphamide (CTX), an alkylating agent, is extensively used in the treatment of lupus nephritis, but its administration has been associated with free radical mediated oxidative stress. The present study was designed to investigate the effect of dietary corn oil (CO), fish oil (FO) and food restriction (FR) on the activities of hepatic antioxidant enzymes, fatty acid composition and lipid peroxidation following CTX administration in autoimmune prone NZB/W female mice. METHODS: Autoimmune-prone NZB/W female mice were fed either ad libitum (AL) or food restricted (60% of AL intake), semipurified diets containing 5% CO or 5% FO supplemented with equal levels of antioxidants and injected with either phosphate buffered saline (PBS), or CTX (50 mg/kg body weight) every 10 days. Proteinuria was measured biweekly. The treatment was stopped at 10 months and diets were continued until the mice were killed at 12 months. Fatty acid composition, activity of antioxidant enzymes and lipid peroxidation were analyzed in liver homogenates, and anti-DNA antibodies were analyzed in the serum. RESULTS: Mice in the FO/AL dietary group exhibited significantly higher liver catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities compared to the CO/AL dietary group. CTX significantly decreased SOD and GSH-Px activity in the FO/AL group and CAT and GSH-Px in the CO/AL group. In AL fed mice given CTX, activities of CAT, GSH Px and GST were significantly higher in mice fed FO diets than in mice fed CO diets. FR increased the activity of enzymes in both the CO and FO diet groups. In FR mice, CTX decreased CAT and GSH-Px activity in both the CO and FO dietary groups, but glutathione S-transferase (GST) only in the CO group. The decrease in SOD activity was not significant in either of the restricted groups. CTX significantly increased generation of thiobarbituric acid reactive substances (TBARS) in both AL groups. FR significantly decreased lipid peroxidation in both the CO and FO groups, with or without CTX. CTX decreased serum anti-DNA antibody levels in both the CO and FO dietary groups. FR also decreased antibody titer in both the CO and FO dietary groups, and it was decreased further with CTX treatment. FO fed animals had higher levels of n-3 fatty acids, whereas CO fed animals had high levels of n-6 fatty acids. CTX significantly increased 20:4 and decreased 18:1 in CO/AL fed animals, whereas it increased 18:1 and decreased 22:6 in FO/AL fed animals. CONCLUSIONS: Results obtained in the present study suggests that FO and, more significantly, FO combined with FR can have a beneficial effect in hepatic tissues subjected to CTX induced oxidative stress by regulating the activity of antioxidant enzymes. In addition, the study also indicates that n-3 and n-6 dietary lipids are susceptible to lipid peroxidation, particularly in the presence of a prooxidant like CTX, and that FR is beneficial in decreasing lipid peroxidation. The study also suggests that FO and CTX can have additive effects in preventing kidney disease in NZB/W mice. PMID- 14559932 TI - Complex multivitamin supplementation improves homocysteine and resistance to LDL C oxidation. AB - OBJECTIVE: We previously reported in an open-label pilot trial that a 24 ingredient multivitamin formula favorably influenced homocysteine concentration and LDL-C oxidation indices following 24 weeks of supplementation. Our current aim was to more thoroughly examine this same formula in a randomized, placebo controlled, clinical study. METHODS: We examined 182 participants for selected plasma vitamin concentrations and clinically relevant variables including homocysteine, lipids and LDL-C oxidation indices at baseline and six months. RESULTS: We found no significant differences between groups for any parameter at baseline. Following six months of vitamin supplementation, we observed elevations in plasma concentrations of vitamin B6 (as pyridoxal 5'-phosphate; PLP), vitamin B12, folate, vitamin C, vitamin E and beta-carotene (p < 0.0001), all of which were significantly greater than respective placebo group changes (p < 0.0001). Homocysteine decreased in the treatment (8.38 +/- 2.9 vs. 6.93 +/- 2.5 micro mol/L; p < 0.0001) and placebo group (8.17 +/- 3.0 vs. 7.42 +/- 2.2 micro mol/L; p < 0.0001) from baseline to six months, respectively, with reductions in the treatment group being greater than placebo (p < 0.008). LDL-C oxidation indices were also improved as LDL-C oxidation rate was decreased (-0.39 micro mol/min/g protein; p < 0.0003) and LDL-C lag time increased (11.3 min; p < 0.003) in supplemented participants. Further analysis also showed that LDL-C oxidation rate was lower (p < 0.0007) and LDL-C lag time longer (p < 0.0001) for the vitamin group than placebo treatment after six months. CONCLUSION: We conclude that a multi-ingredient vitamin formula with antioxidant properties has measurable effects on homocysteine and LDL-C oxidation indices. PMID- 14559933 TI - Fan beam dual energy X-ray absorptiometry body composition measurements in piglets. AB - OBJECTIVES: A piglet model was used to validate and cross validate the fan-beam (FB) dual energy X-ray absorptiometry (DXA) software vKH6 and to determine the predictive values of physiologic parameters (weight, length, age and gender) on body composition. METHODS: Nineteen piglets (Group A: 600 to 21100 g) were used to validate the FB-DXA measurements of body composition based on chemical analysis of the carcass. An additional 22 piglets (Group B: 640 g to 17660 g) had FB-DXA measurements, and these values were compared to the predicted values generated from regression equations computed from group A piglets. Body composition for bone mass, lean mass and fat mass was based on ash weight, nitrogen and fat measured from three aliquots of homogenate from each carcass. Data from all piglets (n = 41) were used to determine the variations in body composition. Data analysis used regression, t test and analysis of variance. RESULTS: Duplicate DXA (total weight TW, bone mineral content BMC, bone area BA, bone mineral density BMD, lean mass LM and fat mass FM) measurements were highly correlated (r = 0.98 to 1.00, p < 0.001 for all comparisons) and were not significantly different. No significant differences were found in the residuals from predicted versus measured DXA values between the larger and the smaller (<1.6 kg) piglets from Group A. For Group B piglets, the DXA measured TW of 5666 +/- 5692 g (mean +/- SD), LM (5063 +/- 5048 g), FM (465 +/- 510 g), BMC (138 +/- 139 g), BA (486 +/- 365 cm(2)) and BMD (0.235 +/- 0.071 g/cm(2)) were highly significantly correlated with (r = 0.94 to 1.00, p < 0.001 for all comparisons) and were not significantly different from the predicted values. Data from all piglets (n = 41) showed that weight is the dominant predictor of whole body and regional body composition. Length, age or gender contributed to <2% of the variability of body composition. CONCLUSION: Body composition measurements using the FB DXA software vKH6 is highly reproducible. The software vKH6 is validated for use in a wide range of body weights and body composition, and cross-validated using a separate group of animals. Body weight is the dominant predictor of body composition in immature piglets. PMID- 14559934 TI - Anti-TNF-alpha antibody normalizes serum leptin in IL-2 deficient mice. AB - OBJECTIVE: A recent study reports that the interleukin-2 deficient (IL-2(-/-)) mouse model of autoimmune and inflammatory bowel disease (IBD) with elevated pro inflammatory cytokine production has elevated leptin concentrations during food deprivation. The objective of this study was to examine whether increased tumor necrosis factor-alpha (TNF-alpha), a pro-inflammatory cytokine, contributes to the abnormally elevated leptin in IL-2(-/-) mice. METHODS: Eight week old, IL-2( /-) and wild-type control (IL-2(+/+)), male mice were fed regular laboratory mouse food for two weeks. At the end of the study, blood was collected in the fed state, IL-2(-/-) and IL-2(+/+) mice were injected with either anti-TNF-alpha monoclonal antibody or normal saline, and blood was collected in the starved state. RESULTS: The IL-2(-/-) mice consumed less food and lost weight. Administration of anti-TNF-alpha antibody markedly reduced serum leptin concentrations in IL-2(-/-) and control mice after food deprivation. Serum leptin in the IL-2(-/-) mice not receiving anti-TNF-alpha antibody increased significantly in the starved state. Serum concentrations of TNF-alpha were higher in IL-2(-/-) mice compared to controls in both the fed and starved state. CONCLUSIONS: These results suggest that elevated TNF-alpha may be one mechanism for the sustained elevated leptin observed in IL-2(-/-) mice during food deprivation. PMID- 14559935 TI - Studying cancer in 3 dimensions: 3-D models foster new insights into tumorigenesis. PMID- 14559936 TI - Gene studies, anti-TNF therapy take Lasker honors. PMID- 14559937 TI - New centers to focus on health disparities. PMID- 14559944 TI - Treatment of depression by mental health specialists and primary care physicians. PMID- 14559945 TI - Treatment of depression by mental health specialists and primary care physicians. PMID- 14559946 TI - Treatment of depression by mental health specialists and primary care physicians. PMID- 14559947 TI - Cost-effectiveness of treatment for chronic hepatitis C infection. PMID- 14559948 TI - Cost-effectiveness of treatment for chronic hepatitis C infection. PMID- 14559949 TI - Prescription drug benefits and health status among Medicare patients. PMID- 14559950 TI - Communicating life expectancy to patients with terminal illness. PMID- 14559951 TI - State legislation and use of feeding tubes in cognitively impaired patients. PMID- 14559952 TI - Processes and outcomes of care among navajo women with breast cancer. PMID- 14559953 TI - Results of therapy for acute lymphoblastic leukemia in black and white children. AB - CONTEXT: Treatment results for acute lymphoblastic leukemia (ALL) clearly have improved over the past decade, but black children have not fared as well as white children in large national trials. OBJECTIVE: To compare the clinical outcomes of therapy for black and white children with ALL treated at a single institution. DESIGN, SETTING, AND PATIENTS: A retrospective analysis of 412 children and adolescents (68 black, 338 white, and 6 other race) with newly diagnosed ALL who were treated consecutively at a pediatric cancer center in Memphis, Tenn. Patients were enrolled from December 1991 to July 1998 in successive Total Therapy studies regardless of race, ethnicity, or ability to pay and received risk-directed therapy according to stringent criteria. INTERVENTIONS: All patients received the same intensive, remission-induction therapy followed by 120 weeks of risk-assigned postremission therapy that included reinduction treatment, pulses of high-dose methotrexate, and early intensification of intrathecal chemotherapy. MAIN OUTCOME MEASURES: Event-free and overall survival rates for black and white children were estimated by the method of Kaplan and Meier and compared with the Mantel-Haenszel test and by Cox proportional hazards regression analysis, adjusting for known prognostic factors. RESULTS: The 68 black children were significantly more likely than the 338 white children to have higher-risk prognostic features, including an initial leukocyte count greater than 100 x 10(3)/ microL, a T-cell immunophenotype, and the t(1;19) chromosomal translocation with E2A-PBX1 fusion, and were less likely to have hyperdiploid blast cells, a favorable prognostic factor in childhood ALL. However, the clinical outcomes for these 2 cohorts were not significantly different: 5-year event-free and overall survival rates were 80.7% (95% confidence interval [CI], 70.3%-91.1%) and 86.2% (95% CI, 77.2%-95.2%) for black children vs 79.4% (95% CI, 74.7%-84.1%) and 85.0% (95% CI, 80.9%-89.1%) for white children. Ten-year results also were comparable, but the CIs were wide because of the small numbers of patients who had been followed up for 10 years or more. The lack of a racial effect on the long-term outcome of therapy was still apparent in a multivariate Cox regression analysis, adjusting for sex, age, presenting leukocyte count, leukemic cell DNA index, immunophenotype, and central nervous system status. CONCLUSION: With equal access to effective antileukemic therapy, black and white children with ALL can expect the same high rate of cure. PMID- 14559954 TI - Survival variability by race and ethnicity in childhood acute lymphoblastic leukemia. AB - CONTEXT: The role of race/ethnicity in survival of children with acute lymphoblastic leukemia (ALL) is unclear, with some studies reporting poorer survival among minority children and others reporting equivalent survival across race/ethnicity in the modern, risk-stratified treatment era. OBJECTIVE: To investigate the relation between race/ethnicity and survival in a large, population-based analysis of incident ALL cases in the United States. DESIGN, POPULATION, AND SETTING: This study included 4952 individuals diagnosed with ALL between 1973 and 1999 at age 19 years or younger. ALL cases were identified from 9 population-based registries of the National Cancer Institute's Surveillance, Epidemiology, and End Results program. MAIN OUTCOME MEASURES: Survival probabilities were compared among white, black, Hispanic, Asian/Pacific Islander, and American Indian/Alaskan Native children. Kaplan-Meier curves and proportional hazard ratios from Cox regression analysis were calculated, accounting for treatment era (1973-1982, 1983-1989, and 1990-1999), age at diagnosis (<1, 1-9, and 10-19 years), and sex. RESULTS: Although overall 5-year survival probabilities improved with each successive treatment era, differences according to race/ethnicity persisted. For 1990-1999, 5-year survival was 84% for white children, 81% for Asian/Pacific Islander children, 75% for black children, and 72% for both American Indian/Alaskan Native children and Hispanic children. The largest difference by race/ethnicity was observed among children diagnosed between ages 1 and 9 years. Compared with white children, after adjusting for treatment era, age at diagnosis, and sex, children of black, Hispanic, and American Indian/Alaskan Native descent had hazard ratios of 1.50 (95% CI, 1.0 2.2; P =.03), 1.83 (95% CI, 1.4-2.4; P<.001), and 1.90 (95% CI, 0.8-4.6; P =.16). CONCLUSIONS: Black, Hispanic, and American Indian/Alaskan Native children with ALL have worse survival than white and Asian/Pacific Islander children, even in the contemporary treatment era. Future work must delineate the social and biological factors, including any differences in pharmacokinetics associated with chemotherapeutic agents, that account for disparities in outcome. PMID- 14559955 TI - Exercise plus behavioral management in patients with Alzheimer disease: a randomized controlled trial. AB - CONTEXT: Exercise training for patients with Alzheimer disease combined with teaching caregivers how to manage behavioral problems may help decrease the frailty and behavioral impairment that are often prevalent in patients with Alzheimer disease. OBJECTIVE: To determine whether a home-based exercise program combined with caregiver training in behavioral management techniques would reduce functional dependence and delay institutionalization among patients with Alzheimer disease. DESIGN, SETTING, AND PATIENTS: Randomized controlled trial of 153 community-dwelling patients meeting National Institute of Neurological and Communicative Diseases and Stroke/Alzheimer Disease and Related Disorders Association criteria for Alzheimer disease, conducted between June 1994 and April 1999. INTERVENTIONS: Patient-caregiver dyads were randomly assigned to the combined exercise and caregiver training program, Reducing Disability in Alzheimer Disease (RDAD), or to routine medical care (RMC). The RDAD program was conducted in the patients' home over 3 months. MAIN OUTCOME MEASURES: Physical health and function (36-item Short-Form Health Survey's [SF-36] physical functioning and physical role functioning subscales and Sickness Impact Profile's Mobility subscale), and affective status (Hamilton Depression Rating Scale and Cornell Depression Scale for Depression in Dementia). RESULTS: At 3 months, in comparison with the routine care patients, more patients in the RDAD group exercised at least 60 min/wk (odds ratio [OR], 2.82; 95% confidence interval [CI], 1.25-6.39; P =.01) and had fewer days of restricted activity (OR, 3.10; 95% CI, 1.08-8.95; P<.001). Patients in the RDAD group also had improved scores for physical role functioning compared with worse scores for patients in the RMC group (mean difference, 19.29; 95% CI, 8.75-29.83; P<.001). Patients in the RDAD group had improved Cornell Depression Scale for Depression in Dementia scores while the patients in the RMC group had worse scores (mean difference, -1.03; 95% CI, -0.17 to -1.91; P =.02). At 2 years, the RDAD patients continued to have better physical role functioning scores than the RMC patients (mean difference, 10.89; 95% CI, 3.62-18.16; P =.003) and showed a trend (19% vs 50%) for less institutionalization due to behavioral disturbance. For patients with higher depression scores at baseline, those in the RDAD group improved significantly more at 3 months on the Hamilton Depression Rating Scale (mean difference, 2.21; 95% CI, 0.22-4.20; P =.04) and maintained that improvement at 24 months (mean difference, 2.14; 95% CI, 0.14-4.17; P =.04). CONCLUSION: Exercise training combined with teaching caregivers behavioral management techniques improved physical health and depression in patients with Alzheimer disease. PMID- 14559956 TI - Relationships between poverty and psychopathology: a natural experiment. AB - CONTEXT: Social causation (adversity and stress) vs social selection (downward mobility from familial liability to mental illness) are competing theories about the origins of mental illness. OBJECTIVE: To test the role of social selection vs social causation of childhood psychopathology using a natural experiment. DESIGN: Quasi-experimental, longitudinal study. POPULATION AND SETTING: A representative population sample of 1420 rural children aged 9 to 13 years at intake were given annual psychiatric assessments for 8 years (1993-2000). One quarter of the sample were American Indian, and the remaining were predominantly white. Halfway through the study, a casino opening on the Indian reservation gave every American Indian an income supplement that increased annually. This increase moved 14% of study families out of poverty, while 53% remained poor, and 32% were never poor. Incomes of non-Indian families were unaffected. MAIN OUTCOME MEASURES: Levels of Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, psychiatric symptoms in the never-poor, persistently poor, and ex-poor children were compared for the 4 years before and after the casino opened. RESULTS: Before the casino opened, the persistently poor and ex-poor children had more psychiatric symptoms (4.38 and 4.28, respectively) than the never-poor children (2.75), but after the opening levels among the ex-poor fell to those of the never poor children, while levels among those who were persistently poor remained high (odds ratio, 1.50; 95% confidence interval, 1.08-2.09; and odds ratio, 0.91; 95% confidence interval, 0.77-1.07, respectively). The effect was specific to symptoms of conduct and oppositional defiant disorders. Anxiety and depression symptoms were unaffected. Similar results were found in non-Indian children whose families moved out of poverty during the same period. CONCLUSIONS: An income intervention that moved families out of poverty for reasons that cannot be ascribed to family characteristics had a major effect on some types of children's psychiatric disorders, but not on others. Results support a social causation explanation for conduct and oppositional disorder, but not for anxiety or depression. PMID- 14559957 TI - Unique lipoprotein phenotype and genotype associated with exceptional longevity. AB - CONTEXT: Individuals with exceptional longevity have a lower incidence and/or significant delay in the onset of age-related disease, and their family members may inherit biological factors that modulate aging processes and disease susceptibility. OBJECTIVE: To identify specific biological and genetic factors that are associated with or reliably define a human longevity phenotype. DESIGN, SETTING, AND PARTICIPANTS: In a case-control design, 213 Ashkenazi Jewish probands with exceptional longevity (mean [SD] age, 98.2 [5.3] years) and their offspring (n = 216; mean [SD] age, 68.3 [6.7] years) were recruited from 1998 to 2002, while an age-matched control group of Ashkenazi Jews (n = 258) and participants from the Framingham Offspring Study (n = 589) were accepted as control groups. MAIN OUTCOME MEASURES: Detailed questionnaires, physical examination, and blood samples were taken, including assessment of lipids and lipoprotein subclass levels and particle sizes by proton nuclear magnetic resonance. Samples were also genotyped for the codon 405 isoleucine to valine (I405V) variation in the cholesteryl ester transfer protein (CETP) gene, which is involved in regulation of lipoprotein and its particle sizes. RESULTS: High density lipoprotein (HDL) and low-density lipoprotein (LDL) particle sizes were significantly higher in probands compared with both control groups (P =.001 for both), independent of plasma levels of HDL and LDL cholesterol and apolipoprotein A1 and B. This phenotype was also typical of the proband's offspring but not of the age-matched controls. The HDL and LDL particle sizes were significantly larger in offspring and controls without hypertension or cardiovascular disease, (P =.001 and P =.008, respectively). Furthermore, lipoprotein particle sizes, but not plasma LDL levels, were significantly higher in offspring and controls without the metabolic syndrome (P<.001). Probands and offspring had a 2.9- and 3.6-fold (in men) and 2.7- and 1.5-fold (in women) increased frequency, respectively, of homozygosity for the 405 valine allele of CETP (VV genotype), respectively, compared with controls (P<.001 for both). Those probands with the VV genotype had increased lipoprotein sizes and lower serum CETP concentrations. CONCLUSIONS: Individuals with exceptional longevity and their offspring have significantly larger HDL and LDL particle sizes. This phenotype is associated with a lower prevalence of hypertension, cardiovascular disease, the metabolic syndrome, and increased homozygosity for the I405V variant in CETP. These findings suggest that lipoprotein particle sizes are heritable and promote a healthy aging phenotype. PMID- 14559958 TI - Glucose control and mortality in critically ill patients. AB - CONTEXT: Hyperglycemia is common in critically ill patients, even in those without diabetes mellitus. Aggressive glycemic control may reduce mortality in this population. However, the relationship between mortality, the control of hyperglycemia, and the administration of exogenous insulin is unclear. OBJECTIVE: To determine whether blood glucose level or quantity of insulin administered is associated with reduced mortality in critically ill patients. DESIGN, SETTING, AND PATIENTS: Single-center, prospective, observational study of 531 patients (median age, 64 years) newly admitted over the first 6 months of 2002 to an adult intensive care unit (ICU) in a UK national referral center for cardiorespiratory surgery and medicine. MAIN OUTCOME MEASURES: The primary end point was intensive care unit (ICU) mortality. Secondary end points were hospital mortality, ICU and hospital length of stay, and predicted threshold glucose level associated with risk of death. RESULTS: Of 531 patients admitted to the ICU, 523 underwent analysis of their glycemic control. Twenty-four-hour control of blood glucose levels was variable. Rates of ICU and hospital mortality were 5.2% and 5.7%, respectively; median lengths of stay were 1.8 (interquartile range, 0.9-3.7) days and 6 (interquartile range, 4.5-8.3) days, respectively. Multivariable logistic regression demonstrated that increased administration of insulin was positively and significantly associated with ICU mortality (odds ratio, 1.02 [95% confidence interval, 1.01-1.04] at a prevailing glucose level of 111-144 mg/dL [6.1-8.0 mmol/L] for a 1-IU/d increase), suggesting that mortality benefits are attributable to glycemic control rather than increased administration of insulin. Also, the regression models suggest that a mortality benefit accrues below a predicted threshold glucose level of 144 to 200 mg/dL (8.0-11.1 mmol/L), with a speculative upper limit of 145 mg/dL (8.0 mmol/L) for the target blood glucose level. CONCLUSIONS: Increased insulin administration is positively associated with death in the ICU regardless of the prevailing blood glucose level. Thus, control of glucose levels rather than of absolute levels of exogenous insulin appear to account for the mortality benefit associated with intensive insulin therapy demonstrated by others. PMID- 14559959 TI - Care of the dying doctor: on the other end of the stethoscope. AB - The challenges of caring for a dying doctor reflect both common issues in helping the terminally ill and unique problems in working with a physician-patient. The dying doctor must deal with a familiar environment and set of problems from a radically different perspective and must negotiate overlapping and conflicting personal and professional roles. Some of the cardinal virtues of physicians- professional identity, expertise, perfectionism, selflessness, and stoicism--may pose both strengths and liabilities in the patient's role. The treating physicians may also encounter new strains in caring for a colleague. They must guard against both overinvolvement and underinvolvement, and, as with all dying persons, they must serve as a guide through unfamiliar territory for dying patient and family--a companion who is not afraid to listen to or explore the most upsetting matters, a person who can speak frankly when others may be ignoring "the horse on the dining room table." The case of Dr B, an internist dying of myelofibrosis and congestive heart failure, whose son is also a physician, offers the reader the opportunity to reflect on these challenges and to draw lessons about how to best care for fellow physicians at a time of great need. We suggest strategies for negotiating the patient-physician relationship when the patient is also a physician. PMID- 14559960 TI - Complexities in prognostication in advanced cancer: "to help them live their lives the way they want to". PMID- 14559961 TI - Important causes of visual impairment in the world today. PMID- 14559962 TI - Race and outcome in childhood acute lymphoblastic leukemia. PMID- 14559963 TI - Poverty and child mental health: natural experiments and social causation. PMID- 14559964 TI - JAMA patient page. Causes of visual impairment. PMID- 14559965 TI - Capture of an intermediate in the catalytic cycle of L-aspartate-beta semialdehyde dehydrogenase. AB - The structural analysis of an enzymatic reaction intermediate affords a unique opportunity to study a catalytic mechanism in extraordinary detail. Here we present the structure of a tetrahedral intermediate in the catalytic cycle of aspartate-beta-semialdehyde dehydrogenase (ASADH) from Haemophilus influenzae at 2.0-A resolution. ASADH is not found in humans, yet its catalytic activity is required for the biosynthesis of essential amino acids in plants and microorganisms. Diaminopimelic acid, also formed by this enzymatic pathway, is an integral component of bacterial cell walls, thus making ASADH an attractive target for the development of new antibiotics. This enzyme is able to capture the substrates aspartate-beta-semialdehyde and phosphate as an active complex that does not complete the catalytic cycle in the absence of NADP. A distinctive binding pocket in which the hemithioacetal oxygen of the bound substrate is stabilized by interaction with a backbone amide group dictates the R stereochemistry of the tetrahedral intermediate. This pocket, reminiscent of the oxyanion hole found in serine proteases, is completed through hydrogen bonding to the bound phosphate substrate. PMID- 14559966 TI - Crystal structure of the tyrosine kinase domain of the hepatocyte growth factor receptor c-Met and its complex with the microbial alkaloid K-252a. AB - The protooncogene c-met codes for the hepatocyte growth factor receptor tyrosine kinase. Binding of its ligand, hepatocyte growth factor/scatter factor, stimulates receptor autophosphorylation, which leads to pleiotropic downstream signaling events in epithelial cells, including cell growth, motility, and invasion. These events are mediated by interaction of cytoplasmic effectors, generally through Src homology 2 (SH2) domains, with two phosphotyrosine containing sequence motifs in the unique C-terminal tail of c-Met (supersite). There is a strong link between aberrant c-Met activity and oncogenesis, which makes this kinase an important cancer drug target. The furanosylated indolocarbazole K-252a belongs to a family of microbial alkaloids that also includes staurosporine. It was recently shown to be a potent inhibitor of c-Met. Here we report the crystal structures of an unphosphorylated c-Met kinase domain harboring a human cancer mutation and its complex with K-252a at 1.8-A resolution. The structure follows the well established architecture of protein kinases. It adopts a unique, inhibitory conformation of the activation loop, a catalytically noncompetent orientation of helix alphaC, and reveals the complete C-terminal docking site. The first SH2-binding motif (1349YVHV) adopts an extended conformation, whereas the second motif (1356YVNV), a binding site for Grb2-SH2, folds as a type II Beta-turn. The intermediate portion of the supersite (1353NATY) assumes a type I Beta-turn conformation as in an Shc-phosphotyrosine binding domain peptide complex. K-252a is bound in the adenosine pocket with an analogous binding mode to those observed in previously reported structures of protein kinases in complex with staurosporine. PMID- 14559967 TI - Adaptive mutation: general mutagenesis is not a programmed response to stress but results from rare coamplification of dinB with lac. AB - In a particular genetic system, selection stimulates reversion of a lac mutation and causes genome-wide mutagenesis (adaptive mutation). Selection allows rare plated cells with a duplication of the leaky lac allele to initiate clones within which further lac amplification improves growth rate. Growth and amplification add mutational targets to each clone and thereby increase the likelihood of reversion. We suggest that general mutagenesis occurs only in clones whose lac amplification includes the nearby dinB+ gene (for error-prone DNA polymerase IV). Thus mutagenesis is not a programmed response to stress but a side effect of amplification in a few clones; it is not central to the effect of selection on reversion. PMID- 14559968 TI - Glia-independent chains of neuroblasts through the subcortical parenchyma of the adult rabbit brain. AB - In the brains of adult mammals long-distance cell migration of neuronal precursors is known to occur in the rostral migratory stream, involving chains of cells sliding into astrocytic glial tubes. By combining immunocytochemistry for polysialylated neural cell adhesion molecule (PSA-NCAM), neuronal and glial antigens, endogenous and exogenously administered cell-proliferation markers, and light and electron microscopy 3D reconstructions, we show that chains of newly generated neuroblasts exist both inside and outside the subventricular zone of adult rabbits. Two groups of chains were detectable within the mature brain parenchyma: anterior chains, into the anterior forceps of the corpus callosum, and posterior chains, close to the external capsule. Parenchymal chains were not associated with any special glial structures, thus coming widely in contact with the mature nervous tissue, including unmyelinated/myelinated fibers, astrocytes, neurons, and oligodendrocytes. These chains of cells, unlike those in the subventricular zone, do not display cell proliferation, but they contain BrdUrd administered several weeks before. Telencephalic areas, such as the putamen, amygdala, claustrum, and cortex, adjacent to the chains harbor numerous PSA-NCAM positive cells. The counting of newly generated cells in these areas shows small differences in comparison with others, and a few cells double-labeled for BrdUrd/PSA-NCAM (after 1-month survival) and for BrdUrd/NeuN (after 2 months) were detectable. These results demonstrate the occurrence of glial-independent chains of migrating neuroblasts, which directly contact the mature brain parenchyma of adult mammals. These chains could provide a possible link between the adult germinative layers and a very low-rate/long-term process of cell addition in the telencephalon. PMID- 14559970 TI - Energetics of hydrogen bonds in peptides. AB - Hydrogen bonds and their relative strengths in proteins are of importance for understanding protein structure and protein motions. The correct strength of such hydrogen bonds is experimentally known to vary greatly from approximately 5-6 kcal/mol for the isolated bond to approximately 0.5-1.5 kcal/mol for proteins in solution. To estimate these bond strengths, here we suggest a direct novel kinetic procedure. This analyzes the timing of the trajectories of a properly averaged dynamic ensemble. Here we study the observed rupture of these hydrogen bonds in a molecular dynamics calculation as an alternative to using thermodynamics. This calculation is performed for the isolated system and contrasted with results for water. We find that the activation energy for the rupture of the hydrogen bond in a beta-sheet under isolated conditions is 4.76 kcal/mol, and the activation energy is 1.58 kcal/mol for the same beta-sheet in water. These results are in excellent agreement with observations and suggest that such a direct calculation can be useful for the prediction of hydrogen bond strengths in various environments of interest. PMID- 14559969 TI - DNA-mediated charge transport for DNA repair. AB - MutY, like many DNA base excision repair enzymes, contains a [4Fe4S]2+ cluster of undetermined function. Electrochemical studies of MutY bound to a DNA-modified gold electrode demonstrate that the [4Fe4S] cluster of MutY can be accessed in a DNA-mediated redox reaction. Although not detectable without DNA, the redox potential of DNA-bound MutY is approximately 275 mV versus NHE, which is characteristic of HiPiP iron proteins. Binding to DNA is thus associated with a change in [4Fe4S]3+/2+ potential, activating the cluster toward oxidation. Given that DNA charge transport chemistry is exquisitely sensitive to perturbations in base pair structure, such as mismatches, we propose that this redox process of MutY bound to DNA exploits DNA charge transport and provides a DNA signaling mechanism to scan for mismatches and lesions in vivo. PMID- 14559971 TI - Principles of cell-free genetic circuit assembly. AB - Cell-free genetic circuit elements were constructed in a transcription translation extract. We engineered transcriptional activation and repression cascades, in which the protein product of each stage is the input required to drive or block the following stage. Although we can find regions of linear response for single stages, cascading to subsequent stages requires working in nonlinear regimes. Substantial time delays and dramatic decreases in output production are incurred with each additional stage because of a bottleneck at the translation machinery. Faster turnover of RNA message can relieve competition between genes and stabilize output against variations in input and parameters. PMID- 14559972 TI - RNA-editing cytidine deaminase Apobec-1 is unable to induce somatic hypermutation in mammalian cells. AB - Antibody diversification by somatic hypermutation, gene conversion, and class switch recombination is completely dependent on activation-induced cytidine deaminase (AID). A recent report showing induction of DNA mutations in Escherichia coli by overexpression of AID, Apobec-1, and related members of the RNA-editing cytidine deaminase family suggested that they may directly modify deoxycytidine in DNA in mammalian cells (DNA-editing model). We therefore examined whether Apobec-1 bona fide RNA-editing enzyme could show somatic hypermutation and class switching activities in murine B lymphocytes and fibroblasts. Unlike AID, Apobec-1 was unable to induce somatic hypermutation or class switching. The results force a reevaluation of the physiological significance of the DNA deaminase activities of AID and Apobec-1 in E. coli and in vitro. PMID- 14559974 TI - Bovine mammary gene expression profiling using a cDNA microarray enhanced for mammary-specific transcripts. AB - A cDNA microarray resource enhanced for transcripts specific to the bovine mammary gland (BMAM) has been developed and used in pilot studies to examine gene expression profiles in the mammary gland. One goal driving development of this resource was to shed some light on the pathways and mechanisms specifically related to bovine mammary gland growth and development. To accomplish this, gene expression patterns from bovine adipose, liver, adrenal, lymph, spleen, thymus, gut, and developing mammary tissue were compared using the BMAM microarray. We have thus identified a putative set of 16 genes being preferentially expressed in developing mammary gland. Another of our long-term goals is to elucidate the genes and pathways associated with bovine lactation and involution and to use these as a model for human mammary gland development as it relates to human breast cancer risks. To begin this process, we conducted a pilot study, comparing gene expression profiles of lactating bovine mammary tissue against nonlactating tissue on the BMAM microarray. Our results have yielded many novel and interesting genes exhibiting differential expression in lactating mammary tissue, including oncogenes (VAV3, C-myc), mediators of apoptosis (Caspase 8), and cell cycle regulators (LASP1). PMID- 14559975 TI - Development of a porcine brain cDNA library, EST database, and microarray resource. AB - Recent developments in expressed sequence tag (EST) and cDNA microarray technology have had a dramatic impact on the ability of scientists to study responses of thousands of genes to internal and external stimuli. In neurobiology, studies of the human brain have been expanding rapidly by use of functional genomics techniques. To enhance these studies and allow use of a porcine brain model, a normalized porcine brain cDNA library (PBL) has been generated and used as a base for EST discovery and microarray generation. In this report, we discuss initial sequence analysis of 965 clones from this resource. Our data revealed that library normalization successfully reduced the number of clones representing highly abundant cDNA species and overall clone redundancy. Cluster analysis revealed over 800 unique cDNA species representing a redundancy rate for the normalized library of 6.9% compared with 29.4% before normalization. Sequence information, BLAST results, and TIGR cluster matches for these ESTs are publicly available via a web-accessible database (http://nbfgc.msu.edu). A cDNA microarray was created using 877 unique porcine brain EST amplicons spotted in triplicate on glass slides. This microarray was assessed by performing a series of experiments designed to test hybridization efficiency and false-positive rate. Our results indicate that the PBL cDNA microarray is a robust tool for studies of brain gene expression using swine as a model system. PMID- 14559973 TI - The contribution of protease-activated receptor 1 to neuronal damage caused by transient focal cerebral ischemia. AB - The serine proteases tissue plasminogen activator, plasmin, and thrombin and their receptors have previously been suggested to contribute to neuronal damage in certain pathological situations. Here we demonstrate that mice lacking protease-activated receptor 1 (PAR1) have a 3.1-fold reduction in infarct volume after transient focal cerebral ischemia. Intracerebroventricular injection of PAR1 antagonist BMS-200261 reduced infarct volume 2.7-fold. There are no detectable differences between PAR1-/- and WT mice in cerebrovascular anatomy, capillary density, or capillary diameter, demonstrating that the neuroprotective phenotype is not likely related to congenital abnormalities in vascular development. We also show that the exogenously applied serine proteases thrombin, plasmin, and tissue plasminogen activator can activate PAR1 signaling in brain tissue. These data together suggest that if blood-derived serine proteases that enter brain tissue in ischemic situations can activate PAR1, this sequence of events may contribute to the harmful effects observed. Furthermore, PAR1 immunoreactivity is present in human brain, suggesting that inhibition of PAR1 may provide a novel potential therapeutic strategy for decreasing neuronal damage associated with ischemia and blood-brain barrier breakdown. PMID- 14559976 TI - Genetic interaction between a maternal factor and the zygotic genome controls the intestine length in PRM/Alf mice. AB - Postoperative management of small and large bowel resections would be helped by use of intestinotrophic molecules. Here, we present a mouse inbred strain called PRM/Alf that is characterized by a selective intestinal lengthening. We show that PRM/Alf intestine is one-third longer compared with other inbred strains. The phenotype is acquired mostly during the postnatal period, before weaning. Its genetic determinism is polygenic, and involves a strong maternal effect. Cross fostering experiments revealed that the dam's genotype acts synergistically with the offspring's genotype to confer the longest intestine. Moreover, genes in the offspring have a direct effect on intestine length. Possible involvement of milk growth factors and identification of candidate genes are discussed. PMID- 14559977 TI - Age-dependent hypothalamic expression of neuropeptides in wild-type and melanocortin-4 receptor-deficient mice. AB - In young (35- to 56-day-old) and middle-aged (9-mo-old) wild-type (+/+) and melanocortin-4 receptor (MC4R)-deficient (+/-, -/-) mice, expressions of neuropeptide Y (NPY), agouti-related protein (AGRP), pro-opiomelanocortin (POMC), and cocaine-and-amphetamine-regulated transcript (CART) were analyzed in the arcuate nucleus (ARC) and adjacent regions comprising the dorsomedial (DMN) and ventromedial (VMN) nucleus. In the ARC of young mice, NPY and AGRP expression increased and POMC and CART expression decreased with body fat content. Adjusting for the influence of body fat content by ANCOVA showed that the levels of NPY, POMC, and CART were highest and of AGRP lowest in young -/- mice. In the middle aged mice, feedback from body fat content was weakened. For -/- mice ANCOVA revealed higher NPY and AGRP, lower POMC, and unchanged CART expression levels relative to young -/- mice. In the DMN and VMN, POMC and AGRP signals were absent at each age. CART was expressed in the DMN independent of age, fat content, and genotype. For NPY expression, an age-dependent induction was found in the DMN and VMN; it was absent in the young but present in the middle-aged mice, showing close positive correlations between body fat content and the numbers of NPY labeled cells which were further enhanced in -/- mice. Thus MC4R deficiency augments age-induced NPY expression in the DMN and VMN with no feedback from body fat content. Negative feedback control by body fat content on ARC neuropeptide expression is present in young animals but vanishes with age and is modulated by MC4R deficiency. PMID- 14559978 TI - Protein gene product 9.5 (PGP 9.5) is not a specific marker of neural and nerve sheath tumors: an immunohistochemical study of 95 mesenchymal neoplasms. AB - In non-neoplastic tissues, the expression of protein gene product 9.5 (PGP 9.5), a member of the ubiquitin hydrolase family of proteins, is confined to neural and neuroendocrine cells. Although it has been claimed that PGP 9.5 is a specific marker of neural and/or nerve sheath differentiation in human tumors, careful review of the literature suggests that relatively few nonneural or nerve sheath tumors have been studied. Prompted by our recent observation of a PGP 9.5 positive malignant fibrous histiocytoma, we undertook a study of PGP 9.5 expression in a large group of well-characterized mesenchymal neoplasms. Sections from 95 mesenchymal tumors were retrieved from our archives and immunostained for PGP 9.5 using standard avidin-biotin complex technique and heat-induced epitope retrieval. Scoring was as follows: negative, 1+ (<10-25% of cells), 2+ (25-50% of cells), and 3+ (>50% of cells). Normal nerves and fibrous tissue were internal positive and negative controls, respectively. Positive immunostaining was seen in 80/95 (84%) of cases. Positive results by tumor subtype were as follows: (1) nerve sheath tumors: malignant peripheral nerve sheath tumor (7/10), neurofibromas (10/10), and perineuriomas (3/3); (2) (Myo) fibroblastic tumors: malignant fibrous histiocytoma (18/20), low-grade fibromyxoid sarcomas (8/9), fibromatoses (7/7), and desmoplastic fibroblastomas (2/2); (3) vascular tumors: angiosarcomas (4/4), hemangioendotheliomas (3/5), and hemangiomas (3/4); and (4) other non-nerve sheath tumors: pleomorphic liposarcoma (4/4), dermatofibrosarcoma protuberans (2/5), rhabdomyosarcomas (2/2), synovial sarcomas (8/8), melanomas (1/2). All positive cases were 2-3+ except 6 malignant peripheral nerve sheath tumor, 1 neurofibroma, 3 malignant fibrous histiocytoma, 2 low-grade fibromyxoid sarcoma, and 1 dermatofibrosarcoma protuberans. Positive staining was seen in normal smooth muscle and germinal centers in addition to nerves. We conclude that in this, the largest study to date of PGP 9.5 expression in mesenchymal neoplasms, we have found strong (2-3+) expression in the vast majority of nonneural or nerve sheath neoplasms studied. Although PGP 9.5 is a sensitive neural/nerve sheath marker, it is essentially totally nonspecific for diagnostic purposes. It is possible that our findings reflect cross-reactivity of the 13C4 clone with epitopes present on other ubiquitin hydrolases. Alternatively, PGP 9.5 expression may be aberrantly up-regulated in a variety of mesenchymal neoplasms. PMID- 14559979 TI - Allelic loss and tumor pathology in head and neck squamous cell carcinoma. AB - Allelic loss is a common occurrence in head and neck tumors and has been shown to be an independent predictor of prognosis; however, the relationship between allelic loss and tumor pathology is not well-known. We studied 139 patients who were newly diagnosed with squamous cell cancer of the head and neck to determine whether tumor pathology was correlated with allelic loss at one or more of eight different regions on chromosomes 3p, 5q, 8p, 9p, 10p, 18q, and 21q. At each chromosomal region, loss of heterozygosity at any one of three or four highly polymorphic microsatellite markers that spanned the region in question was considered evidence for allelic loss. A pathologist scored all tumors for seven tumor pathology and host interface parameters. Mean allelic loss across all eight regions was associated with mitotic index (P =.034) and inflammatory response (P =.005). For allelic loss at specific chromosomal regions, the most statistically significant trends were between overall tumor grade and 3p14.2-p13 (P =.014), mitotic index and 3p24.3-p14.3 (P =.026), 9p24.2-p21 (P =.004) and 18q12.3-q23 (P =.009), inflammatory response and 3p14.2-p13 (P =.008) and 9p24.2-p21 (P =.001), desmoplastic response and 9p24.2-p21 (P =.009), and pattern of invasion and 21q21 q22.2 (P =.015). Our results suggest that genes involved in tumor suppression and oncogenesis can potentially be classified based on specific pathologic events in head and neck squamous cell carcinogenesis that they modify. PMID- 14559980 TI - Pediatric intracranial ependymomas: prognostic relevance of histological, immunohistochemical, and flow cytometric factors. AB - The correlation between the histological features and clinical outcome remains poor in pediatric intracranial ependymomas. We performed a retrospective study of a group of 31 patients (diagnosed from 1985 to 1995) to assess prognostic implications of the current grading system, of histological and immunohistochemical features, and of ploidy status estimated by flow cytometry. Immunoexpression of a broad spectrum of antigens was evaluated, including MIB-1, topoisomerase-IIalpha, cyclin D1, glial and epithelial proteins (GFAP, EMA, cytokeratins), molecules involved in controlling apoptosis (bcl-2, caspase 3/CPP32), and p53 oncoprotein. Univariate and multivariate statistical analyses were performed to evaluate the influence of each variable on both the progression free survival (PFS) and the overall survival (OS) with at least 7-year follow up. Although we showed a significant correlation between histological grade and prognosis, the current grading system failed in predicting outcome in nearly one third of individual cases. Problems with interpathologist reproducibility were also demonstrated. The extent of surgical resection was the only clinical factor that was associated with survival. Both the PFS and the OS were significantly decreased for the following pathological variables: increased cellularity (>300 nuclei per HPF), mitotic activity of >7 per 10 HPF, increased MIB-1 labeling index (LI), topoisomerase-IIalpha LI, S-phase fraction, and p53 and bcl-2 positivity. Increased cyclin D1 LI was demonstrated to have only a marginally significant impact on PFS. A flow chart modeling was further performed to formulate a scheme for discriminating of prognostic subgroups. Based on that, p53 immunopositivity and/or MIB-1 LI of >5% (after subtotal resection) or MIB-1 LI of >15% (after complete resection) were the strongest indicators of the tumor's aggressive behavior and of a poor prognosis of the disease. Foci of hypercellularity should be specifically looked for in ependymomas for assessing the immunohistochemical studies. PMID- 14559982 TI - Clinicopathological correlates of pancreatic intraepithelial neoplasia: a comparative analysis of 82 cases with and 152 cases without pancreatic ductal adenocarcinoma. AB - Pancreatic intraepithelial neoplasia is often associated with pancreatic ductal adenocarcinoma and is presumed to be its precursor. It has been difficult to determine the frequency of these lesions because until recently, there was no consensus regarding the terminology and criteria for their grading. Here we compare the frequency and clinical correlates of pancreatic intraepithelial neoplasia in pancreata involved by ductal adenocarcinoma and in benign ones, using the criteria put forward recently. We evaluated pancreatectomy specimens from 82 patients with ductal adenocarcinoma and 152 patients who underwent pancreatectomy for reasons other than primary malignancy (trauma, pancreatitis, and metastatic tumor to pancreas) for the presence, grade, and number of foci of pancreatic intraepithelial neoplasia. Cases were graded by the highest grade of pancreatic intraepithelial neoplasia focus identified. An average of 5.3 sections of pancreas was available for evaluation (range, 1-28 sections). Overall, the frequency of pancreatic intraepithelial neoplasia lesions in ductal adenocarcinoma patients, including Grade 1A (mucinous duct lesions), was 82%, which was significantly higher than the one in benign pancreata -54%, P <.001. There was a progressive increase from normal pancreata to pancreatitis and to ductal adenocarcinoma in the frequency of overall pancreatic intraepithelial neoplasia lesions (16%, 60%, and 82%, respectively) and Grade 3 pancreatic intraepithelial neoplasia (0%, 4%, and 40%, respectively). In most instances, in any given case of higher-grade pancreatic intraepithelial neoplasia lesion, there were also several foci of lower grade lesions. The frequency of higher-grade pancreatic intraepithelial neoplasia lesions (2 and 3) in pancreata resected for ductal adenocarcinoma was 59%, significantly higher than in those without primary carcinoma (17%). This progressive increase in frequency of pancreatic intraepithelial neoplasia from incidental pancreatectomies (presumed to have a nonpathologic pancreas) to pancreatitis (considered a risk factor for carcinoma) and to ductal adenocarcinoma constitutes an indirect support for the precancerous role attributed to pancreatic intraepithelial neoplasia lesions. The relatively high absolute occurrence of pancreatic intraepithelial neoplasia Grade 1A (mucinous duct lesions) in benign conditions (43%) suggests that this group represents a combination of neoplastic and non-neoplastic lesions. PMID- 14559981 TI - Loss of heterozygosity in the MXI1 gene is a frequent occurrence in melanoma. AB - Melanoma development and progression is thought to be the result of a multi-step accumulation of genetic damage, with loss of heterozygosity in chromosome 9p (MTS1) frequently described. In addition, chromosome 10q allelic loss has been reported, implicating the tumor suppressor gene PTEN/MMAC1 on 10q23.3. The MXI1 gene at 10q24-25 is another candidate tumor suppressor that has only rarely been studied in melanomas, with conflicting results. We used microdissection-based genotyping to investigate 29 melanomas from 20 patients for loss of heterozygosity in intragenic and flanking microsatellite markers for this latter gene. Concurrently, the MTS1 gene was similarly studied using two flanking microsatellites. Fifty-four percent (15 of 28) of the informative cases showed loss of heterozygosity for one or both MXI1 markers, as compared with 67% (16 of 24) of the informative cases for MTS1. MXI1 allelic loss was seen more frequently in recurrent/metastatic tumors (59%), as compared with in primary (33%) lesions. Eighty percent of the primary tumors showed loss of heterozygosity for MTS1, as well as 63% of recurrent/metastatic ones. We studied more than one tumor in eight patients, with those from three patients showing discordant genetic patterns. One patient showed a metastatic tumor with allelic loss for MXI1 that was not identified in the primary melanoma or a local recurrence. The other two patients showed clonal heterogeneity in MXI1 at synchronous and metachronous metastatic foci. These findings support MXI1 as a putative tumor suppressor gene involved in conventional melanoma progression. Genetic heterogeneity seen in different metastases from the same primary suggests a nonlinear pattern of chromosomal damage, with the development of multiple clones within the primary tumor, each acquiring its own metastatic potential. PMID- 14559983 TI - Tumour angiogenesis and p53 protein expression in mammary phyllodes tumors. AB - We examined 186 phyllodes tumors (106 benign, 51 borderline, 29 malignant) for angiogenesis by assessing stromal microvessel density by the hot spot method and assessing p53 protein expression; we correlated these factors with stromal cellularity, margin status, nuclear pleomorphism, mitosis, and stromal overgrowth. Increased degree of malignancy in phyllodes tumors is associated with increased patient age and tumor size. Microvessel density and p53 protein expression also showed a similar increase with malignancy. Using a logistic regression model, microvessel density was shown to be useful in predicting malignancy in phyllodes tumors, independent of key criteria of stromal overgrowth, nuclear pleomorphism, and mitosis. Microvessel density showed correlation with stromal cellularity and margin status, suggesting an interrelationship between these parameters. P53 protein expression showed a positive correlation with microvessel density, suggesting possible overlap in the underlying mechanism of these two factors in the pathogenesis of phyllodes tumors. The numbers of recurrences and metastases are small in our series, and no significant difference was demonstrated in microvessel density and p53 protein expression compared with the primary. We conclude that microvessel density and p53 are useful as independent criteria in evaluating malignancy in phyllodes tumors. PMID- 14559984 TI - E-cadherin promoter hypermethylation in preneoplastic and neoplastic skin lesions. AB - E-cadherin is a calcium-dependent, intercellular adhesion molecule that is specifically expressed in epithelial tissues and plays an important role in maintaining epithelial stability. E-cadherin is widely regarded as a prognostic marker in many types of human cancers. The inactivation of the E-cadherin gene is linked to increased potential for tumor invasiveness and distant metastasis. We previously demonstrated reduced expression of E-cadherin protein immunohistochemically in invasive squamous cell carcinomas of the skin as compared with adjacent normal skin. An epigenetic alteration in association with promoter hypermethylation is one important mechanism of gene silencing. In the present study, we analyze the E-cadherin gene promoter hypermethylation in preneoplastic and neoplastic skin lesions to determine whether epigenetic alteration of the E-cadherin gene also plays an important role in cutaneous squamous carcinogenesis. A total of 33 cases was examined for evidence of E cadherin promoter hypermethylation, and these consist of nine cases of spongiotic dermatitis as nonneoplastic skin control, nine cases of actinic keratosis, eight cases of squamous cell carcinoma in situ, and seven cases of invasive squamous cell carcinoma. Promoter hypermethylation of the E-cadherin gene was detected in 6 of 7 cases (85%) of invasive squamous cell carcinoma, 4 of 8 cases (50%) of squamous cell carcinoma in situ, 4 of 9 cases (44%) of actinic keratosis, and 2 of 9 cases (22%) of nonneoplastic skin. We conclude that E-cadherin promoter hypermethylation occurs frequently and may represent an important mechanism of E cadherin inactivation in cutaneous preneoplastic and neoplastic lesions. The frequencies of E-cadherin promoter hypermethylation appear to be correlated with more advanced stage of squamous carcinogenesis in skin. PMID- 14559985 TI - Tenascin expression at the invasive front is associated with poor prognosis in intrahepatic cholangiocarcinoma. AB - Tenascin and decorin are components of the extracellular matrix (ECM) that are implicated in cell proliferation in tumors. Here, we propose that abnormal expression of stromal ECM may play an important role in the progression of intrahepatic cholangiocarcinoma, which is characterized by desmoplastic reaction. To explore this hypothesis, we performed immunohistochemical analysis in order to examine the expression and distribution of tenascin and decorin in 75 cases of intrahepatic cholangiocarcinoma. In the intratumoral stroma, positive staining for tenascin was observed in 51 (68%) cases, and positive staining for decorin was observed in 61 (81%) cases. However, at the invasive front, positive staining for tenascin was found in 23 (31%) cases, and positive staining for decorin was found in 6 (8%) cases. Decorin staining was not correlated with aggressive behavior of intrahepatic cholangiocarcinoma, whereas intratumoral tenascin staining was correlated with lymphatic permeation and proliferative activity measured by Ki67. Tenascin staining at the invasive front was associated with tumor size, lymphatic permeation, lymph node metastasis, and proliferative activity and appeared to be a useful prognostic factor by univariate analysis, although it was not an independent prognostic factor. These results indicate that tenascin plays a role in tumor progression in cases of intrahepatic cholangiocarcinoma and that tenascin expression, especially at the invasive front, may be a useful marker in evaluating an unfavorable prognosis in patients with intrahepatic cholangiocarcinoma. PMID- 14559986 TI - CD9 expression on lymphatic vessels in head and neck mucosa. AB - CD9, a member of the transmembrane 4 superfamily, is involved in cell adhesion, migration, and tumor metastasis. Little is known about its vascular expression pattern. In this study, we investigated CD9 expression on endothelial cells in the mucosa of the head and neck and compared it with vascular tumors. Using immunohistochemistry, expression of CD9 was studied in 17 samples of head and neck mucosa and skin (laryngeal mucosa: n = 2, oral: n = 6, and epidermis: n = 9) and a variety of vascular tumors (lymphangiomas: n = 9, juvenile nasopharyngeal angiofibromas: n = 4, hemangiomas: n = 7, angiosarcomas: n = 5, and Kaposi's sarcomas: n = 7) and compared with the expression of CD34 and PAL-E (blood vessel markers) and the lymphatic marker podoplanin. Regular lymphatic endothelium and lymphangiomas were strongly positive for CD9 and podoplanin but were mostly negative for PAL-E and CD34. By contrast, blood vessel endothelium and hemangiomas were strongly positive for PAL-E and CD34 but were mostly negative for CD9 and podoplanin. Weak to moderate CD9 reactivity was also observed on EC of juvenile nasopharyngeal angiofibromas, angiosarcomas, and Kaposi's sarcomas. Expression of CD9 by lymphatic EC was confirmed by reverse-transcriptase PCR and Western blot analyses. CD9 may be useful as a marker for lymphatic EC. It could promote the adherence of inflammatory and tumor cells to lymphatic EC and participate in the growth and maintenance of the lymphatic capillary net. PMID- 14559987 TI - Expression of HER2 and C-KIT in nasopharyngeal carcinoma: implications for a new therapeutic approach. AB - We sought to determine the expression and prognostic significance of HER2 and c KIT proteins in nasopharyngeal carcinoma (NPC). In this retrospective study, immunohistochemical stains for HER2 and c-KIT were performed on formalin-fixed paraffin-embedded sections from 49 patients with NPC who were treated at our hospital from 1971 to 2000. The clinical and immunohistochemical data were correlated, including gender, ethnic origin, age, histological type, EBV status (EBER in situ hybridization), stage, and overall survival. HER2 expression was not found in the tested samples. C-KIT overexpression was found in 33% (16/49) of the patients. Nine of the 16 samples (56%) were strongly positive for c-KIT protein (staining of >50% of the tumor cells). C-KIT expression was associated with younger age. C-KIT was not found in patients with squamous carcinoma or in those with negative EBV status, although these two groups consisted of only five patients each. Although c-KIT-positive cases tended to be associated with slightly better survival, this was not statistically significant. C-KIT protein was expressed in one third of the NPC patients in this study, only in EBV positive, undifferentiated, or nonkeratinizing carcinoma patients. Further study is needed to check whether c-KIT expression is correlated with c-KIT DNA mutations and to test the possibility of treatment with imatinib mesylate (Gleevec). HER2 protein was negative in the same tested specimens. PMID- 14559988 TI - Kit expression in small cell carcinomas of the lung: effects of chemotherapy. AB - A significant number of small cell lung carcinomas shows overexpression of the proto-oncogene c-kit product, a tyrosine kinase known as Kit or CD117. This molecular pathway seems somewhat implicated in promoting the neoplastic growth of small cell lung carcinoma. The current pharmacological availability of its selective inhibitor, together with the promising clinical results in the management of CD117-positive neoplasms such as advanced gastrointestinal stromal tumors, aroused great interest among oncologists in also adopting this therapeutic strategy in other CD117-positive tumors. We evaluated a series of 27 small cell lung carcinomas, comparing the expression of CD117 of the primary naive tumor (before first-line chemotherapy) with the expression of the same neoplasm after postchemotherapy relapse. All the patients underwent similar chemotherapeutic regimens (cisplatin/carboplatin plus etoposide). At diagnosis, 21 of 27 cases (78%) showed strong immunoreactivity for CD117. Among these 21 originally positive tumors, CD117 remained overexpressed in 10 after relapse (48%), whereas the other 11 cases became negative. No originally CD117-negative small cell carcinomas displayed immunoreactivity after chemotherapy. CD117 expression was not statistically correlated with overall survival, occurrence of chemoresistance, or clinical response to chemotherapy. We also evaluated CD117 expression in a series of 46 surgically resected non-small cell lung carcinomas (8 squamous cell carcinomas, 10 adenocarcinomas, 5 pleomorphic carcinomas, 10 typical and 3 atypical carcinoids, and 10 large cell neuroendocrine carcinomas). Apart from small cell carcinomas, CD117 overexpression was observed in 6 of 10 large cell neuroendocrine carcinomas, whereas all the other histotypes resulted unstained. We speculate that loss of CD117 expression after chemotherapy in a high proportion of SCLC indicates that in this tumor, Kit unlikely represents the product of a constitutive mutation, as instead shown in gastrointestinal stromal tumors. Keeping this finding in mind, oncologists could re-test CD117 expression in relapsing small cell lung carcinomas in order to establish the best candidates for enrollment in ongoing clinical trials with Kit inhibitors. Practically speaking, CD117 may be helpful in discriminating between pulmonary high-grade neuroendocrine tumors and other histotypes, but pathologists should be aware that treated small cell lung carcinomas may remain unstained in a not insignificant number of cases. PMID- 14559989 TI - Frameshift mutations in the bax gene are not involved in development of ovarian endometrioid carcinoma. AB - The purpose of this study was to determine whether mutations in the Bax gene play a role in the development of ovarian endometrioid carcinoma with a microsatellite instability phenotype. We analyzed a total of 60 tumor specimens, 49 ovarian endometrioid carcinomas and 11 concurrent endometrial endometrioid carcinomas from 49 patients. Fourteen ovarian endometrioid carcinomas and 6 endometrial endometrioid carcinomas showed a microsatellite instability-high phenotype. Tumor and normal-tissue specimens from eight patients with a microsatellite instability high phenotype colorectal carcinoma were included in this study as controls. The presence or absence of a mutation in the poly (G) 8 tract of the Bax gene was determined by polymerase chain reaction followed by direct DNA sequence analysis. A 1-base pair deletion at the poly (G) 8 tract and no expression of Bax and Bcl-2 proteins were identified in one microsatellite instability-high endometrial endometrioid carcinoma. Immunohistochemical staining for Bax and Bcl-2 proteins was negative on the tumor specimen that had this 1-base pair deletion. No mutations were found in the synchronous microsatellite instability-high ovarian endometrioid carcinoma from the same patient. In contrast, four (50%) of the eight microsatellite instability-high sporadic colorectal carcinomas had a mutation in the poly (G) 8 tract. Although Bax plays an important role in carcinogenesis of the colorectum with microsatellite instability-high phenotype, Bax may not play a direct role in the genesis of ovarian endometrioid carcinoma, regardless of microsatellite instability status. PMID- 14559990 TI - Gains of chromosomes 7, 17, 12, 16, and 20 and loss of Y occur early in the evolution of papillary renal cell neoplasia: a fluorescent in situ hybridization study. AB - It has been suggested that gains of chromosomes 7 and 17 and loss of Y occur in renal papillary adenoma and that progression to papillary renal cell carcinoma is marked by gains of additional chromosomes, most frequently 12, 16, and 20. Previous studies have included very few lesions of <5 mm in diameter, a requirement of the present definition of papillary adenoma. Ten papillary adenomas (ranging from 1 to 5 mm in diameter) from autopsy material and 10 surgically resected papillary renal cell carcinomas were studied with fluorescence in situ hybridization in paraffin sections using centromeric probes for chromosomes 7, 12, 16, 17, 20, and Y diluted 1:100 with tDenHyb1 buffer. The signals in 50 to 150 nuclei were counted in each tumor. Controls for all the probes were normal renal tissues from the same patients. Three or more signals per nucleus were frequently observed in papillary adenomas: chromosome 7 (range, 10 to 50%; > or = 30% in 9 of 10), 17 (range, 10 to 47%; > or = 30% in 7), 16 (range, 1 to 63%; > or = 10% in 5), 12 (range, 0 to 32%; > or =10% in 4), and 20 (range, 5 to 49%; > or = 10% in 5). Loss of the Y chromosome was observed in 80 to 90% of nuclei in 9 adenomas from males. Three or more signals were frequent in papillary renal cell carcinomas: chromosome 7 (range, 32 to 63%; > or =30% in 10 of 10), 17 (range, 28 to 61%; > or = 30% in 7), 16 (range, 0 to 45%; > or = 10% in 6), 12 (range, 1 to 37, > or = 10% in 5), 20 (range, 2 to 44%; > or = 10% in 4). No signal for Y was observed in 12 to 88% (> or = 81% in 6) of nuclei in 7 carcinomas from males. Statistical analysis showed no difference between adenomas and carcinomas. Gains of chromosomes 7, 17, 16, 12, and 20 and loss of the Y chromosome occur early in the evolution of papillary renal cell neoplasia in tumors that are only a few millimeters in diameter. Progressive gains of these chromosomes do not appear to correlate with the transition from adenoma to carcinoma. PMID- 14559991 TI - Metanephric adenoma lacks the gains of chromosomes 7 and 17 and loss of Y that are typical of papillary renal cell carcinoma and papillary adenoma. AB - Metanephric adenoma has morphologic similarities to papillary renal cell neoplasms. Cytogenetic studies of papillary renal cell carcinoma and papillary adenoma have shown frequent gains of chromosomes 7 and 17 and loss of the Y chromosome. Some cytogenetic studies have supported the hypothesis that metanephric adenoma is related to papillary renal cell neoplasia; others have not. Seven metanephric adenomas were studied with fluorescence in situ hybridization in paraffin sections using centromeric probes for chromosomes 7, 17, and Y diluted 1:100 with tDenHyb1 buffer. The signals in 100 to 200 nuclei were counted in each tumor. Samples of histologically normal renal cortical tubule epithelium were used as controls. In all seven metanephric adenomas, the results for chromosomes 7 and 17 were similar: a high percentage of nuclei with two signals (range, 75 to 85%; median, 79%). Normal kidney showed similar results (range, 78 to 88%; median, 84%). The Y chromosome was present in all three of the tumors from males (range, 86 to 89% of nuclei; median, 87%). Normal kidney gave similar results (range 82% to 91%, median 84%). The presence of chromosomes 7, 17, and Y in metanephric adenomas is similar to their presence in normal kidney. Metanephric adenoma lacks the frequent gains of chromosomes 7 and 17 and losses of the Y chromosome that are typical of papillary renal cell neoplasms, supporting the notion that metanephric adenoma is not related to papillary renal cell carcinoma and papillary adenoma. Genetic analysis of chromosomes 7, 17, and Y may facilitate discrimination of metanephric adenoma from papillary renal cell carcinoma in difficult cases. PMID- 14559993 TI - Regulation of alternative splicing by SRrp86 and its interacting proteins. AB - SRrp86 is a unique member of the SR protein superfamily containing one RNA recognition motif and two serine-arginine (SR)-rich domains separated by an unusual glutamic acid-lysine (EK)-rich region. Previously, we showed that SRrp86 could regulate alternative splicing by both positively and negatively modulating the activity of other SR proteins and that the unique EK domain could inhibit both constitutive and alternative splicing. These functions were most consistent with the model in which SRrp86 functions by interacting with and thereby modulating the activity of target proteins. To identify the specific proteins that interact with SRrp86, we used a yeast two-hybrid library screen and immunoprecipitation coupled to mass spectrometry. We show that SRrp86 interacts with all of the core SR proteins, as well as a subset of other splicing regulatory proteins, including SAF-B, hnRNP G, YB-1, and p72. In contrast to previous results that showed activation of SRp20 by SRrp86, we now show that SAF B, hnRNP G, and 9G8 all antagonize the activity of SRrp86. Overall, we conclude that not only does SRrp86 regulate SR protein activity but that it is, in turn, regulated by other splicing factors to control alternative splice site selection. PMID- 14559994 TI - XBP-1 regulates a subset of endoplasmic reticulum resident chaperone genes in the unfolded protein response. AB - The mammalian unfolded protein response (UPR) protects the cell against the stress of misfolded proteins in the endoplasmic reticulum (ER). We have investigated here the contribution of the UPR transcription factors XBP-1, ATF6alpha, and ATF6beta to UPR target gene expression. Gene profiling of cell lines lacking these factors yielded several XBP-1-dependent UPR target genes, all of which appear to act in the ER. These included the DnaJ/Hsp40-like genes, p58(IPK), ERdj4, and HEDJ, as well as EDEM, protein disulfide isomerase-P5, and ribosome-associated membrane protein 4 (RAMP4), whereas expression of BiP was only modestly dependent on XBP-1. Surprisingly, given previous reports that enforced expression of ATF6alpha induced a subset of UPR target genes, cells deficient in ATF6alpha, ATF6beta, or both had minimal defects in upregulating UPR target genes by gene profiling analysis, suggesting the presence of compensatory mechanism(s) for ATF6 in the UPR. Since cells lacking both XBP-1 and ATF6alpha had significantly impaired induction of select UPR target genes and ERSE reporter activation, XBP-1 and ATF6alpha may serve partially redundant functions. No UPR target genes that required ATF6beta were identified, nor, in contrast to XBP-1 and ATF6alpha, did the activity of the UPRE or ERSE promoters require ATF6beta, suggesting a minor role for it during the UPR. Collectively, these results suggest that the IRE1/XBP-1 pathway is required for efficient protein folding, maturation, and degradation in the ER and imply the existence of subsets of UPR target genes as defined by their dependence on XBP-1. Further, our observations suggest the existence of additional, as-yet-unknown, key regulators of the UPR. PMID- 14559995 TI - PU.1 and pRB interact and cooperate to repress GATA-1 and block erythroid differentiation. AB - PU.1 and GATA-1 are two hematopoietic specific transcription factors that play key roles in development of the myeloid and erythroid lineages, respectively. The two proteins bind to one another and inhibit each other's function in transcriptional activation and promotion of their respective differentiation programs. This mutual antagonism may be an important aspect of lineage commitment decisions. PU.1 can also act as an oncoprotein since deregulated expression of PU.1 in erythroid precursors causes erythroleukemias in mice. Studies of cultured mouse erythroleukemia cell lines indicate that one aspect of PU.1 function in erythroleukemogenesis is its ability to block erythroid differentiation by repressing GATA-1 (N. Rekhtman, F. Radparvar, T. Evans, and A. I. Skoultchi, Genes Dev. 13:1398-1411, 1999). We have investigated the mechanism of PU.1 mediated repression of GATA-1. We report here that PU.1 binds to GATA-1 on DNA. We localized the repression activity of PU.1 to a small acidic N-terminal domain that interacts with the C pocket of pRB, a well-known transcriptional corepressor. Repression of GATA-1 by PU.1 requires pRB, and pRB colocalizes with PU.1 and GATA-1 at repressed GATA-1 target genes. PU.1 and pRB also cooperate to block erythroid differentiation. Our results suggest that one of the mechanisms by which PU.1 antagonizes GATA-1 is by binding to it at GATA-1 target genes and tethering to these sites a corepressor that blocks transcriptional activity and thereby erythroid differentiation. PMID- 14559997 TI - Chk1 kinase negatively regulates mitotic function of Cdc25A phosphatase through 14-3-3 binding. AB - The order and fidelity of cell cycle events in mammals is intimately linked to the integrity of the Chk1 kinase-Cdc25A phosphatase pathway. Chk1 phosphorylation targets Cdc25A for destruction and, as shown here, inhibits interactions between Cdc25A and its mitotic substrate cyclin B1-Cdk1. Phosphorylation of Cdc25A on serine 178 and threonine 507 facilitates 14-3-3 binding, and Chk1 phosphorylates both residues in vitro. Mutation of T507 to alanine (T507A) enhanced the biological activity of Cdc25A. Cdc25A(T507A) was more efficient in binding to cyclin B1, activating cyclin B1-Cdk1, and promoting premature entry into mitosis. We propose that the Chk1/Cdc25A/14-3-3 pathway functions to prevent cells from entering into mitosis prior to replicating their genomes to ensure the fidelity of the cell division process. PMID- 14559996 TI - mSin3A/histone deacetylase 2- and PRMT5-containing Brg1 complex is involved in transcriptional repression of the Myc target gene cad. AB - The role of hSWI/SNF complexes in transcriptional activation is well characterized; however, little is known about their function in transcriptional repression. We have previously shown that subunits of the mSin3A/histone deacetylase 2 (HDAC2) corepressor complex copurify with hSWI/SNF complexes. Here we show that the type II arginine-specific methyltransferase PRMT5, which is involved in cyclin E repression, can be found in association with Brg1 and hBrm based hSWI/SNF complexes. We also show that hSWI/SNF-associated PRMT5 can methylate hypoacetylated histones H3 and H4 more efficiently than hyperacetylated histones H3 and H4. Protein-protein interaction studies indicate that PRMT5 and mSin3A interact with the same hSWI/SNF subunits as those targeted by c-Myc. These observations prompted us to examine the expression profile of the c-Myc target genes, carbamoyl-phosphate synthase-aspartate carbamoyltransferase-dihydroorotase (cad) and nucleolin (nuc). We found that cad repression is altered in cells that express inactive Brg1 and in cells treated with the HDAC inhibitor depsipeptide. Using chromatin immunoprecipitation assays, we found that Brg1, mSin3A, HDAC2, and PRMT5 are directly recruited to the cad promoter. These results suggest that hSWI/SNF complexes, through their ability to interact with activator and repressor proteins, control expression of genes involved in cell growth and proliferation. PMID- 14559998 TI - Modulation of retinoid signaling by a cytoplasmic viral protein via sequestration of Sp110b, a potent transcriptional corepressor of retinoic acid receptor, from the nucleus. AB - Hepatitis C virus (HCV) core protein (core) plays a significant role in the development of chronic liver diseases caused by HCV infection. We have discovered that the core sensitized all-trans-retinoic acid (ATRA)-induced cell death in MCF 7 cells. Activation of retinoic acid receptor alpha (RARalpha)-mediated transcription by the core was also seen in all the cell lines tested. By use of a yeast two-hybrid system, we identified Sp110b as a candidate for a core interacting cellular factor. Although the function of Sp110b has remained unknown, we observed that Sp110b interacts with RARalpha and suppresses RARalpha mediated transcription. These data suggest that Sp110b is a transcriptional cofactor negatively regulating RARalpha-mediated transcription. RNA interference mediated reduction of endogenous Sp110b levels depressed the ability of the core to activate RARalpha-mediated transcription, suggesting an essential role for Sp110b in this pathway. The normal nuclear subcellular localization of Sp110b was altered by molecular interaction with the core to the cytoplasmic surface of the endoplasmic reticulum. This evidence suggests a model in which the core sequesters Sp110b from the nucleus and inactivates its corepressor function to activate RARalpha-mediated transcription. These findings likely describe a novel system in which a cytoplasmic viral protein regulates host cell transcription. PMID- 14559999 TI - Role of the insulin-like growth factor I/insulin receptor substrate 1 axis in Rad51 trafficking and DNA repair by homologous recombination. AB - The receptor for insulin-like growth factor I (IGF-IR) controls normal and pathological growth of cells. DNA repair pathways represent an unexplored target through which the IGF-IR signaling system might support pathological growth leading to cellular transformation. However, this study demonstrates that IGF-I stimulation supports homologous recombination-directed DNA repair (HRR). This effect involves an interaction between Rad51 and the major IGF-IR signaling molecule, insulin receptor substrate 1 (IRS-1). The binding occurs within the cytoplasm, engages the N-terminal domain of IRS-1, and is attenuated by IGF-I mediated IRS-1 tyrosine phosphorylation. In the absence of IGF-I stimulation, or if mutated IGF-IR fails to phosphorylate IRS-1, localization of Rad51 to the sites of damaged DNA is diminished. These results point to a direct role of IRS-1 in HRR and suggest a novel role for the IGF-IR/IRS-1 axis in supporting the stability of the genome. PMID- 14560000 TI - Accelerated fat absorption in intestinal alkaline phosphatase knockout mice. AB - Intestinal alkaline phosphatase (IAP) is the most ancestral of the tissue specific members of the AP gene family. Several studies have suggested an absorptive function for IAP, but in vivo data to this effect have been lacking. We inactivated the mouse IAP gene in embryo-derived stem cells and generated mice homozygous for the null mutation. The mice were macroscopically and histologically normal and fertile and showed no difference from the wild-type controls under normal laboratory conditions. However, when maintained long-term on a high-fat diet, the IAP-deficient mice showed faster body weight gain than did control animals. Histological examination revealed an accelerated transport of fat droplets through the intestinal epithelium and elevation of serum triglyceride levels in the IAP-deficient mice compared to wild-type mice. Our study suggests that IAP participates in a rate-limiting step regulating fat absorption. PMID- 14560001 TI - Role of nod2 in the response of macrophages to toll-like receptor agonists. AB - Nod2 (CARD15) is a macrophage-specific protein containing two CARD domains, a large nucleotide binding domain and leucine-rich repeats. Human genetic studies have linked mutations in NOD2/CARD15 with Crohn's disease, although the mechanisms involved are unknown. However, Nod2 has been proposed to directly bind bacterial lipopolysaccharide (LPS) and subsequently act as an activator of NF kappaB via the association of the CARD domains with Rip2/RICK/CARDIAK. This is hypothesized to constitute a pathogen recognition pathway distinct from Toll-like receptor 4-mediated recognition of LPS. Using targeted mutagenesis, we introduced a mutation to delete the CARD domains of mouse Nod2. Mice lacking Nod2 were indistinguishable from controls and showed no signs of intestinal pathology. Macrophages responded normally to multiple Toll-like receptor agonists in terms of NF-kappaB target activation, mitogen-activated protein kinase activation, and cytokine secretion. However, Nod2(-/-) mice were significantly protected in endotoxin challenge experiments, and Nod2(-/-) macrophages were refractory to muramyl dipeptide stimulation. These results argue that Nod2 does not play an essential, nonredundant role in the response of macrophages to bacterial products but rather plays unexpected roles in regulating systemic responses to pathogens. PMID- 14560002 TI - Impaired regulation of tumor suppressor p53 caused by mutations in the xeroderma pigmentosum DDB2 gene: mutual regulatory interactions between p48(DDB2) and p53. AB - Tumor suppressor p53 controls cell cycle progression and apoptosis following DNA damage, thus minimizing carcinogenesis. Mutations in the human DDB2 gene generate the E subgroup of xeroderma pigmentosum (XP-E). We report here that XP-E strains are defective in UV irradiation-induced apoptosis due to severely reduced basal and UV-induced p53 levels. These defects are restored by infection with a p53 cDNA expression construct or with a DDB2 expression construct if and only if it contains intron 4, which includes a nonmutated p53 consensus-binding site. We propose that both before and after UV irradiation, DDB2 directly regulates p53 levels, while DDB2 expression is itself regulated by p53. PMID- 14560003 TI - Aggregation of expanded polyglutamine domain in yeast leads to defects in endocytosis. AB - The role of aggregation of abnormal proteins in cellular toxicity is of general importance for understanding many neurological disorders. Here, using a yeast model, we demonstrate that mutations in many proteins involved in endocytosis and actin function dramatically enhance the toxic effect of polypeptides with an expanded polyglutamine (polyQ) domain. This enhanced cytotoxicity required polyQ aggregation and was dependent on the yeast protein Rnq1 in its prion form. In wild-type cells, expression of expanded polyQ followed by its aggregation led to specific and acute inhibition of endocytosis, which preceded growth inhibition. Some components of the endocytic machinery were efficiently recruited into the polyQ aggregates. Furthermore, in cells with polyQ aggregates, cortical actin patches were delocalized and actin was recruited into the polyQ aggregates. Aggregation of polyQ in mammalian HEK293 cells also led to defects in endocytosis. Therefore, it appears that inhibition of endocytosis is a direct consequence of polyQ aggregation and could significantly contribute to cytotoxicity. PMID- 14560004 TI - Pressure-induced differential regulation of the two tryptophan permeases Tat1 and Tat2 by ubiquitin ligase Rsp5 and its binding proteins, Bul1 and Bul2. AB - Tryptophan uptake appears to be the Achilles' heel in yeast physiology, since under a variety of seemingly diverse toxic conditions, it becomes the limiting factor for cell growth. When growing cells of Saccharomyces cerevisiae are subjected to high hydrostatic pressure, tryptophan uptake is down-regulated, leading to cell cycle arrest in the G(1) phase. Here we present evidence that the two tryptophan permeases Tat1 and Tat2 are differentially regulated by Rsp5 ubiquitin ligase in response to high hydrostatic pressure. Analysis of high pressure growth mutants revealed that the HPG1 gene was allelic to RSP5. The HPG1 mutation or the bul1Delta bul2Delta double mutation caused a marked increase in the steady-state level of Tat2 but not of Tat1, although both permeases were degraded at high pressure in an Rsp5-dependent manner. There were marked differences in subcellular localization. Tat1 localized predominantly in the plasma membrane, whereas Tat2 was abundant in the internal membranes. Moreover, Tat1 was associated with lipid rafts, whereas Tat2 localized in bulk lipids. Surprisingly, Tat2 became associated with lipid rafts upon the occurrence of a ubiquitination defect. These results suggest that ubiquitination is an important determinant of the localization and regulation of these tryptophan permeases. Determination of the activation volume (DeltaV( not equal )) for Tat1- and Tat2 mediated tryptophan uptake (89.3 and 50.8 ml/mol, respectively) revealed that both permeases are highly sensitive to membrane perturbation and that Tat1 rather than Tat2 is likely to undergo a dramatic conformational change during tryptophan import. We suggest that hydrostatic pressure is a unique tool for elucidating the dynamics of integral membrane protein functions as well as for probing lipid microenvironments where they localize. PMID- 14560005 TI - Identification of a TAL1 target gene reveals a positive role for the LIM domain binding protein Ldb1 in erythroid gene expression and differentiation. AB - The TAL1 (or SCL) gene, originally identified from its involvement by a recurrent chromosomal translocation, encodes a basic helix-loop-helix transcription factor essential for erythropoiesis. Although presumed to regulate transcription, its target genes are largely unknown. We show here that a nuclear complex containing TAL1, its DNA-binding partner E47, zinc finger transcription factor GATA-1, LIM domain protein LMO2, and LIM domain-binding protein Ldb1 transactivates the protein 4.2 (P4.2) gene through two E box GATA elements in its proximal promoter. Binding of this complex to DNA was dependent on the integrity of both E box and GATA sites and was demonstrated to occur on the P4.2 promoter in cells. Maximal transcription in transiently transfected cells required both E box GATA elements and expression of all five components of the complex. This complex was shown, in addition, to be capable of linking in solution double-stranded oligonucleotides corresponding to the two P4.2 E box GATA elements. This DNA-linking activity required Ldb1 and increased with dimethyl sulfoxide-induced differentiation of murine erythroleukemia (MEL) cells. In contrast, enforced expression in MEL cells of dimerization-defective mutant Ldb1, as well as wild-type Ldb1, significantly decreased E box GATA DNA-binding activities, P4.2 promoter activity, and accumulation of P4.2 and beta-globin mRNAs. These studies define a physiologic target for a TAL1- and GATA-1-containing ternary complex and reveal a positive role for Ldb1 in erythroid gene expression and differentiation. PMID- 14560006 TI - Fibroblast growth factor 2-mediated translational control of IAPs blocks mitochondrial release of Smac/DIABLO and apoptosis in small cell lung cancer cells. AB - The mitochondrial release of cytochrome c and Smac/DIABLO has been implicated in the activation of apoptosis in response to cell stress. Smac promotes cytochrome c-induced activation of caspases by sequestering the inhibitor of apoptosis protein (IAP) family of potent caspase suppressors. Differential release from mitochondria of cytochrome c and Smac can occur, but the underlying mechanism and physiological significance of this are unclear. Here we show that the mechanism by which fibroblast growth factor 2 (FGF-2) protects small cell lung cancer (SCLC) cells from etoposide-induced cell death involves inhibition of Smac release but not of cytochrome c release. This process is MEK dependent and correlates with an increased expression of XIAP and cellular IAP-1, mediated principally through translational regulation. Exogenous expression of XIAP is sufficient to inhibit caspase 9 activation, Smac release, and cell death induced by etoposide. Prevention of the FGF-2-promoted increase in levels of functional IAPs by RNA interference or the cell-permeant Smac amino-terminal peptide blocked FGF-2-induced protection. FGF-2 can thus protect SCLC cells from chemotherapeutic drugs by modulating IAP levels via posttranscriptional regulation, providing a mechanism for postmitochondrial survival signaling by the MEK/mitogen-activated protein kinase pathway. PMID- 14560007 TI - Quantitative analysis of CBP- and P300-induced histone acetylations in vivo using native chromatin. AB - In vivo, histone tails are involved in numerous interactions, including those with DNA, adjacent histones, and other, nonhistone proteins. The amino termini are also the substrates for a number of enzymes, including histone acetyltransferases (HATs), histone deacetylases, and histone methyltransferases. Traditional biochemical approaches defining the substrate specificity profiles of HATs have been performed using purified histone tails, recombinant histones, or purified mononucleosomes as substrates. It is clear that the in vivo presentation of the substrate cannot be accurately represented by using these in vitro approaches. Because of the difficulty in translating in vitro results into in vivo situations, we developed a novel single-cell HAT assay that provides quantitative measurements of endogenous HAT activity. The HAT assay is performed under in vivo conditions by using the native chromatin structure as the physiological substrate. The assay combines the spatial resolving power of laser scanning confocal microscopy with simple statistical analyses to characterize CREB binding protein (CBP)- and P300-induced changes in global histone acetylation levels at specific lysine residues. Here we show that CBP and P300 exhibit unique substrate specificity profiles, consistent with the developmental and functional differences between the two HATs. PMID- 14560008 TI - Transcription factor and polymerase recruitment, modification, and movement on dhsp70 in vivo in the minutes following heat shock. AB - The uninduced Drosophila hsp70 gene is poised for rapid activation. Here we examine the rapid changes upon heat shock in levels and location of heat shock factor (HSF), RNA polymerase II (Pol II) and its phosphorylated forms, and the Pol II kinase P-TEFb on hsp70 in vivo by using both real-time PCR assays of chromatin immunoprecipitates and polytene chromosome immunofluorescence. These studies capture Pol II recruitment and progression along hsp70 and reveal distinct spatial and temporal patterns of serine 2 and serine 5 phosphorylation: in uninduced cells, the promoter-paused Pol II shows Ser5 but not Ser2 phosphorylation, and in induced cells the relative level of Ser2-P Pol II is lower at the promoter than at regions downstream. An early time point of heat shock activation captures unphosphorylated Pol II recruited to the promoter prior to P-TEFb, and during the first wave of transcription Pol II and the P-TEFb kinase can be seen tracking together across hsp70 with indistinguishable kinetics. Pol II distributions on several other genes with paused Pol II show a pattern of Ser5 and Ser2 phosphorylation similar to that of hsp70. These studies of factor choreography set important limits in modeling transcription regulatory mechanisms. PMID- 14560009 TI - Egr family members regulate nonlymphoid expression of Fas ligand, TRAIL, and tumor necrosis factor during immune responses. AB - The Fas ligand (FasL)/Fas pathway is crucial for homeostasis of the immune system and peripheral tolerance. Peripheral lymphocyte deletion involves FasL/Fas in at least two ways: coexpression of both Fas and its ligand on T cells, leading to activation-induced cell death, and expression of FasL by nonlymphoid cells, such as intestinal epithelial cells (IEC), that kill Fas-positive T cells. We demonstrate here that superantigen Staphylococcus enterotoxin B (SEB) induced a dramatic upregulation of FasL, TRAIL, and TNF mRNA expression and function in IEC from BALB/c and C57BL/6 mice. Using adoptive transfer in which CD4(+) T cells from OT-2 T-cell receptor transgenic mice were transferred into recipients, we observed an induction in IEC of FasL, TRAIL, and TNF mRNA after administration of antigen. Specific Egr-binding sites have been identified in the 5' promoter region of the FasL gene, and Egr-1, Egr-2, and Egr-3 mRNA in IEC from mice treated with SEB and from transgenic OT-2 mice after administration of antigen was upregulated. Overexpression of Egr-2 and Egr-3 induced endogenous ligand upregulation that was inhibited by overexpression of Egr-specific inhibitor Nab1. These results support a role for Egr family members in nonlymphoid expression of FasL, TRAIL, and TNF. PMID- 14560010 TI - Miz1 is required for early embryonic development during gastrulation. AB - Miz1 is a member of the POZ domain/zinc finger transcription factor family. In vivo, Miz1 forms a complex with the Myc oncoprotein and recruits Myc to core promoter elements. Myc represses transcription through Miz1 binding sites. We now show that the Miz1 gene is ubiquitously expressed during mouse embryogenesis. In order to elucidate the physiological function of Miz1, we have deleted the mouse Miz1 gene by homologous recombination. Miz1(+/-) mice are indistinguishable from wild-type animals; in contrast, Miz1(-/-) embryos are not viable. They are severely retarded in early embryonic development and do not undergo normal gastrulation. Expression of Goosecoid and Brachyury is detectable in Miz1(-/-) embryos, suggesting that Miz1 is not required for signal transduction by Nodal. Expression of p21Cip1, a target gene of Miz1 is unaltered; in contrast, expression of p57Kip2, another target gene of Miz1 is absent in Miz1(-/-) embryos. Miz1(-/-) embryos succumb to massive apoptosis of ectodermal cells around day 7.5 of embryonic development. Our results show that Miz1 is required for early embryonic development during gastrulation. PMID- 14560011 TI - Transcriptional coactivator Cited2 induces Bmi1 and Mel18 and controls fibroblast proliferation via Ink4a/ARF. AB - Cited2 (CBP/p300 interacting transactivator with ED-rich tail 2) is required for embryonic development, coactivation of transcription factor AP-2, and inhibition of hypoxia-inducible factor 1 transactivation. Cited2 is induced by multiple growth factors and cytokines and oncogenically transforms cells. Here, we show that the proliferation of Cited2(-/-) mouse embryonic fibroblasts ceases prematurely. This is associated with a reduction in growth fraction, senescent cellular morphology, and increased expression of the cell proliferation inhibitors p16(INK4a), p19(ARF), and p15(INK4b). Deletion of INK4a/ARF (encoding p16(INK4a) and p19(ARF)) completely rescued the defective proliferation of Cited2(-/-) fibroblasts. However, the deletion of INK4a/ARF did not rescue the embryonic malformations observed in Cited2(-/-) mice, indicating that INK4a/ARF independent pathways are likely to be involved here. We found that Cited2(-/-) fibroblasts had reduced expression of the polycomb-group genes Bmi1 and Mel18, which function as INK4a/ARF and Hox repressors. Complementation with CITED2 expressing retrovirus enhanced proliferation, induced Bmi1/Mel18 expression, and decreased INK4a/ARF expression. Bmi1- and Mel18-expressing retroviruses enhanced the proliferation of Cited2(-/-) fibroblasts, indicating that they function downstream of Cited2. Our results provide genetic evidence that Cited2 controls the expression of INK4a/ARF and fibroblast proliferation, at least in part via the polycomb-group genes Bmi1 and Mel18. PMID- 14560012 TI - Critical role of Ser-520 phosphorylation for membrane recruitment and activation of the ZAP-70 tyrosine kinase in T cells. AB - Regulation of protein tyrosine kinases (PTKs) by tyrosine phosphorylation is well recognized; in fact, nearly all PTKs require phosphorylation of tyrosine residues in their "activation loop" for catalytic activity. In contrast, the phosphorylation of PTKs on serine and threonine residues has not been studied nearly as much. We report that the ZAP-70 PTK contains predominately phosphoserine in normal T lymphocytes as well as in Jurkat T leukemia cells. We have identified one site of phosphorylation as Ser-520 and find this site to be important for the recruitment and activation of ZAP-70 in T cells. Mutant ZAP-70 S520A had reduced ability to autophosphorylate and to mediate antigen receptor induced interleukin 2 gene activation and was not enriched at the plasma membrane. These defects were rescued by addition of a myristylation signal to the N terminus of ZAP-70-S520A to force its plasma membrane and lipid raft localization. We conclude that phosphorylation of ZAP-70 at Ser-520 plays an important role in the correct localization of ZAP-70 and in priming ZAP-70 for its acute recruitment and activation upon antigen receptor ligation. PMID- 14560013 TI - Separation of retinoid X receptor homo- and heterodimerization functions. AB - As a promiscuous dimerization partner the retinoid X receptor (RXR) can contribute to signaling by multiple nuclear receptors. However, the impact of RXR cosignaling and the possible existence of an RXR homodimer signaling pathway are largely unexplored. We report here on the separation of RXR homo- and heterodimerization as an essential step towards the elucidation of the roles of RXR homo- and heterodimers in retinoid-rexinoid signaling. RXR homodimerization was specifically disrupted by single mutations in the RXR dimerization interface. In contrast, even multiple mutations did not fully impair RXR heterodimerization with retinoic acid receptor (RAR). Importantly, the mutation of mouse RXRalpha (mRXRalpha) Tyr402 substantially weakened RAR heterodimerization while concomitantly increasing homodimerization. Not only did this lead to cooperatively enhanced RXR homodimer binding to DR1 or DR5 elements, but unexpectedly, the mutant acquired significant binding efficiency for noncognate DR3 or DR4 elements as well. The increased stability of RXR homodimers on DR1 correlated with increased transcriptional activity of mRXRalpha(Y402A) on DR1 based reporter genes. Weak, if any, heterodimerization was observed with thyroid, vitamin D(3), or peroxisome proliferator-activating receptors. A model accounting for the structural impact of the Tyr402 mutation on dimerization is discussed. These results provide the basis for a genetic replacement of wild-type RXRs by mutants like mRXRalpha(Y402A) to elucidate the physiological impact of RXR homo- and heterodimerization. PMID- 14560014 TI - Human artificial chromosomes with alpha satellite-based de novo centromeres show increased frequency of nondisjunction and anaphase lag. AB - Human artificial chromosomes have been used to model requirements for human chromosome segregation and to explore the nature of sequences competent for centromere function. Normal human centromeres require specialized chromatin that consists of alpha satellite DNA complexed with epigenetically modified histones and centromere-specific proteins. While several types of alpha satellite DNA have been used to assemble de novo centromeres in artificial chromosome assays, the extent to which they fully recapitulate normal centromere function has not been explored. Here, we have used two kinds of alpha satellite DNA, DXZ1 (from the X chromosome) and D17Z1 (from chromosome 17), to generate human artificial chromosomes. Although artificial chromosomes are mitotically stable over many months in culture, when we examined their segregation in individual cell divisions using an anaphase assay, artificial chromosomes exhibited more segregation errors than natural human chromosomes (P < 0.001). Naturally occurring, but abnormal small ring chromosomes derived from chromosome 17 and the X chromosome also missegregate more than normal chromosomes, implicating overall chromosome size and/or structure in the fidelity of chromosome segregation. As different artificial chromosomes missegregate over a fivefold range, the data suggest that variable centromeric DNA content and/or epigenetic assembly can influence the mitotic behavior of artificial chromosomes. PMID- 14560015 TI - Caenorhabditis elegans EVL-14/PDS-5 and SCC-3 are essential for sister chromatid cohesion in meiosis and mitosis. AB - Sister chromatid cohesion is fundamental for the faithful transmission of chromosomes during both meiosis and mitosis. Proteins involved in this process are highly conserved from yeasts to humans. In screenings for sterile animals with abnormal vulval morphology, mutations in the Caenorhabditis elegans evl-14 and scc-3 genes were isolated. Defects in cell divisions were observed in germ line as well as in vulval and somatic gonad lineages. Through positional cloning of these genes, we have shown that EVL-14 and SCC-3 are likely the only C. elegans homologs of the yeast sister chromatid cohesion proteins Pds5 and Scc3, respectively. Both evl-14 and scc-3 mutants displayed defects in the meiotic germ line. In evl-14 mutants, synaptonemal complexes (SCs) were detectable but more than the usual six DAPI (4',6'-diamidino-2-phenylindole)-positive structures were seen at diakinesis, suggesting that EVL-14/PDS-5 is important for the maintenance of sister chromatid cohesion in late prophase. In scc-3 mutant animals, normal SCs were not visible and approximately 24 DAPI-positive structures were seen at diakinesis, indicating that SCC-3 is necessary for sister chromatid cohesion. Immunostaining revealed that localization of REC-8, a homolog of the yeast meiotic cohesin subunit Rec8, to the chromosomes depends on the presence of SCC-3 but not that of EVL-14/PDS-5. scc-3 RNA interference (RNAi)-treated embryos were 100% lethal and displayed defects in cell divisions. evl-14 RNAi caused a range of phenotypes. These results indicate that EVL-14/PDS-5 and SCC-3 have functions in both mitosis and meiosis. PMID- 14560016 TI - Cbl-3-deficient mice exhibit normal epithelial development. AB - Cbl family proteins are evolutionarily conserved ubiquitin ligases that negatively regulate signaling from tyrosine kinase-coupled receptors. The mammalian cbl family consists of c-Cbl, Cbl-b, and the recently cloned Cbl-3 (also known as Cbl-c). In this study, we describe the detailed expression pattern of murine Cbl-3 and report the generation and characterization of Cbl-3-deficient mice. Cbl-3 exhibits an expression pattern distinct from those of c-Cbl and Cbl b, with high levels of Cbl-3 expression in epithelial cells of the gastrointestinal tract and epidermis, as well as the respiratory, urinary, and reproductive systems. Cbl-3 expression was not detected in nonepithelial cells, but within epithelial tissues, the levels of Cbl-3 expression varied from undetectable in the alveoli of the lungs to very strong in the cecum and colon. Despite this restricted expression pattern, Cbl-3-deficient mice were viable, healthy, and fertile and displayed no histological abnormalities up to 18 months of age. Proliferation of epithelial cells in the epidermises and gastrointestinal tracts was unaffected by the loss of Cbl-3. Moreover, Cbl-3 was not required for attenuation of epidermal growth factor-stimulated Erk activation in primary keratinocytes. Thus, Cbl-3 is dispensable for normal epithelial development and function. PMID- 14560018 TI - Elimination of protein kinase MK5/PRAK activity by targeted homologous recombination. AB - MK5 (mitogen-activated protein kinase [MAPK]-activated protein kinase 5), also designated PRAK (p38-regulated and -activated kinase), was deleted from mice by homologous recombination. Although no MK5 full-length protein and kinase activity was detected in the MK5 knockout mice, the animals were viable and fertile and did not display abnormalities in tissue morphology or behavior. In addition, these mice did not show increased resistance to endotoxic shock or decreased lipopolysaccharide-induced cytokine production. Hence, MK5 deletion resulted in a phenotype very different from the complex inflammation-impaired phenotype of mice deficient in MK2, although MK2 and MK5 exhibit evolutional, structural, and apparent extensive functional similarities. To explain this discrepancy, we used wild-type cells and embryonic fibroblasts from both MK2 and MK5 knockout mice as controls to reexamine the mechanism of activation, the interaction with endogenous p38 MAPK, and the substrate specificity of both enzymes. In contrast to MK2, which shows interaction with and chaperoning properties for p38 MAPK and which is activated by extracellular stresses such as arsenite or sorbitol treatment, endogenous MK5 did not show these properties. Furthermore, endogenous MK5 is not able to phosphorylate Hsp27 in vitro and in vivo. We conclude that the differences between the phenotypes of MK5- and MK2-deficient mice result from clearly different functional properties of both enzymes. PMID- 14560017 TI - Histone deacetylation of RB-responsive promoters: requisite for specific gene repression but dispensable for cell cycle inhibition. AB - The retinoblastoma tumor suppressor protein (RB) is targeted for inactivation in the majority of human tumors, underscoring its critical role in attenuating cellular proliferation. RB inhibits proliferation by repressing the transcription of genes that are essential for cell cycle progression. To repress transcription, RB assembles multiprotein complexes containing chromatin-modifying enzymes, including histone deacetylases (HDACs). However, the extent to which HDACs participate in transcriptional repression and are required for RB-mediated repression has not been established. Here, we investigated the role of HDACs in RB-dependent cell cycle inhibition and transcriptional repression. We find that active RB mediates histone deacetylation on cyclin A, Cdc2, topoisomerase IIalpha, and thymidylate synthase promoters. We also demonstrate that this deacetylation is HDAC dependent, since the HDAC inhibitor trichostatin A (TSA) prevented histone deacetylation at each promoter. However, TSA treatment blocked RB repression of only a specific subset of genes, thereby demonstrating that the requirement of HDACs for RB-mediated transcriptional repression is promoter specific. The HDAC-independent repression was not associated with DNA methylation or gene silencing but was readily reversible. We show that this form of repression resulted in altered chromatin structure and was dependent on SWI/SNF chromatin remodeling activity. Importantly, we find that cell cycle inhibitory action of RB is not intrinsically dependent on the ability to recruit HDAC activity. Thus, while HDACs do play a major role in RB-mediated repression, they are dispensable for the repression of critical targets leading to cell cycle arrest. PMID- 14560019 TI - Role of the steroid receptor coactivator SRC-3 in cell growth. AB - Steroid receptor coactivator 3 (SRC-3/p/CIP/AIB1/ACTR/RAC3/TRAM-1) is a member of the p160 family of nuclear receptor coactivators, which includes SRC-1 (NCoA-1) and SRC-2 (TIF2/GRIP1/NCoA2). Previous studies indicate that SRC-3 is required for normal animal growth and is often amplified or overexpressed in many cancers, including breast and prostate cancers. However, the mechanisms of SRC-3-mediated growth regulation remain unclear. In this study, we show that overexpression of SRC-3 stimulates cell growth to increase cell size in prostate cancer cell lines. Furthermore, our results indicate that overexpression of SRC-3 can modulate the AKT signaling pathway in a steroid-independent manner, which results in the activation of AKT/mTOR signaling concomitant with an increase in cell size. In contrast, down-regulation of SRC-3 expression in cells by small interfering RNA decreases cell growth, leading to a smaller cell size. Similarly, in SRC-3 null mutant mice, AKT signaling is down-regulated in normally SRC-3-expressing tissues. Taken together, these results suggest that SRC-3 is an important modulator for mammalian cell growth. PMID- 14560020 TI - Retinoic acid receptor-mediated induction of ABCA1 in macrophages. AB - ABCA1, the mutant molecule in Tangier Disease, mediates efflux of cellular cholesterol to apoA-I and is induced by liver X receptor (LXR)/retinoid X receptor (RXR) transcription factors. Retinoic acid receptor (RAR) activators (all-trans-retinoic acid [ATRA] and TTNPB) were found to increase ATP-binding cassette transporter 1 (ABCA1) mRNA and protein in macrophages. In cellular cotransfection assays, RARgamma/RXR activated the human ABCA1 promoter, via the same direct repeat 4 (DR4) promoter element as LXR/RXR. Chromatin immunoprecipitation analysis in macrophages confirmed the binding of RARgamma/RXR to the ABCA1 promoter DR4 element in the presence of ATRA, with weaker binding of RARalpha/RXR, and no binding of RARbeta/RXR. However, in macrophages from RARgamma(-/-) mice, TTNPB still induced ABCA1, in association with marked upregulation of RARalpha, suggesting that high levels of RARalpha can compensate for the absence of RARgamma. Dose-response experiments with ATRA in mouse primary macrophages showed that other LXR target genes were weakly induced (ABCG1 and SREBP-1c) or not induced (apoE and LXRalpha). The more specific RAR activator TTNPB did not induce SREBP-1c in mouse primary macrophages or liver. These studies indicate a direct role of RARgamma/RXR in induction of macrophage ABCA1. PMID- 14560021 TI - Dynamic properties of nucleosomes during thermal and ATP-driven mobilization. AB - The fundamental subunit of chromatin, the nucleosome, is not a static entity but can move along DNA via either thermal or enzyme-driven movements. Here we have monitored the movements of nucleosomes following deposition at well-defined locations on mouse mammary tumor virus promoter DNA. We found that the sites to which nucleosomes are deposited during chromatin assembly differ from those favored during thermal equilibration. Taking advantage of this, we were able to track the movement of nucleosomes over 156 bp and found that this proceeds via intermediate positions spaced between 46 and 62 bp. The remodeling enzyme ISWI was found to direct the movement of nucleosomes to sites related to those observed during thermal mobilization. In contrast, nucleosome mobilization driven by the SWI/SNF and RSC complexes were found to drive nucleosomes towards sites up to 51 bp beyond DNA ends, with little respect for the sites favored during thermal repositioning. The dynamic properties of nucleosomes we describe are likely to influence their role in gene regulation. PMID- 14560023 TI - Ceramide disables 3-phosphoinositide binding to the pleckstrin homology domain of protein kinase B (PKB)/Akt by a PKCzeta-dependent mechanism. AB - Ceramide is generated in response to numerous stress-inducing stimuli and has been implicated in the regulation of diverse cellular responses, including cell death, differentiation, and insulin sensitivity. Recent evidence indicates that ceramide may regulate these responses by inhibiting the stimulus-mediated activation of protein kinase B (PKB), a key determinant of cell fate and insulin action. Here we show that inhibition of this kinase involves atypical PKCzeta, which physically interacts with PKB in unstimulated cells. Insulin reduces the PKB-PKCzeta interaction and stimulates PKB. However, dissociation of the kinase complex and the attendant hormonal activation of PKB were prevented by ceramide. Under these circumstances, ceramide activated PKCzeta, leading to phosphorylation of the PKB-PH domain on Thr(34). This phosphorylation inhibited phosphatidylinositol 3,4,5-trisphosphate (PIP(3)) binding to PKB, thereby preventing activation of the kinase by insulin. In contrast, a PKB-PH domain with a T34A mutation retained the ability to bind PIP(3) even in the presence of a ceramide-activated PKCzeta and, as such, expression of PKB T34A mutant in L6 cells was resistant to inhibition by ceramide treatment. Inhibitors of PKCzeta and a kinase-dead PKCzeta both antagonized the inhibitory effect of ceramide on PKB. Since PKB confers a prosurvival signal and regulates numerous pathways in response to insulin, suppressing its activation by a PKCzeta-dependent process may be one mechanism by which ceramide promotes cell death and induces insulin resistance. PMID- 14560022 TI - Identification of NAP1, a regulatory subunit of IkappaB kinase-related kinases that potentiates NF-kappaB signaling. AB - The IkappaB kinase (IKK)-related kinase NAK (also known as TBK or T2K) contributes to the activation of NF-kappaB-dependent gene expression. Here we identify NAP1 (for NAK-associated protein 1), a protein that interacts with NAK and its relative IKK epsilon (also known as IKKi). NAP1 activates NAK and facilitates its oligomerization. Interestingly, the NAK-NAP1 complex itself effectively phosphorylated serine 536 of the p65/RelA subunit of NF-kappaB, and this activity was stimulated by tumor necrosis factor alpha (TNF-alpha). Overexpression of NAP1 specifically enhanced cytokine induction of an NF-kappaB dependent, but not an AP-1-dependent, reporter. Depletion of NAP1 reduced NF kappaB-dependent reporter gene expression and sensitized cells to TNF-alpha induced apoptosis. These results define NAP1 as an activator of IKK-related kinases and suggest that the NAK-NAP1 complex may protect cells from TNF-alpha induced apoptosis by promoting NF-kappaB activation. PMID- 14560024 TI - Post-TATA binding protein recruitment clearance of Gcn5-dependent histone acetylation within promoter nucleosomes. AB - Transcriptional activation of eukaryotic genes often requires the function of histone acetyltransferases (HATs), which is expected to result in the hyperacetylation of histones within promoter nucleosomes. In this study we show that, in Saccharomyces cerevisiae, the steady-state levels of Gcn5-dependent histone acetylation within a number of transcriptionally active promoters are inversely related to the rate of transcription. High acetylation levels were measured only when transcription was attenuated either by TATA element mutations or in a strain carrying a temperature-sensitive protein component of RNA polymerase II. In addition, we show that in one case the low levels of histone acetylation depend on the function of the Rpd3 histone deacetylase. These results point to the existence of an unexpected interplay of two opposing histone modifying activities which operate on promoter nucleosomes following the initiation of RNA synthesis. Such interplay could ensure rapid turnover of chromatin acetylation states in continuously reprogrammed transcriptional systems. PMID- 14560025 TI - Mitochondria use different mechanisms for transport of multispanning membrane proteins through the intermembrane space. AB - The mitochondrial inner membrane contains numerous multispanning integral proteins. The precursors of these hydrophobic proteins are synthesized in the cytosol and therefore have to cross the mitochondrial outer membrane and intermembrane space to reach the inner membrane. While the import pathways of noncleavable multispanning proteins, such as the metabolite carriers, have been characterized in detail by the generation of translocation intermediates, little is known about the mechanism by which cleavable preproteins of multispanning proteins, such as Oxa1, are transferred from the outer membrane to the inner membrane. We have identified a translocation intermediate of the Oxa1 preprotein in the translocase of the outer membrane (TOM) and found that there are differences from the import mechanisms of carrier proteins. The intermembrane space domain of the receptor Tom22 supports the stabilization of the Oxa1 intermediate. Transfer of the Oxa1 preprotein to the inner membrane is not affected by inactivation of the soluble TIM complexes. Both the inner membrane potential and matrix heat shock protein 70 are essential to release the preprotein from the TOM complex, suggesting a close functional cooperation of the TOM complex and the presequence translocase of the inner membrane. We conclude that mitochondria employ different mechanisms for translocation of multispanning proteins across the aqueous intermembrane space. PMID- 14560026 TI - Diarylurea compounds inhibit caspase activation by preventing the formation of the active 700-kilodalton apoptosome complex. AB - The release of mitochondrial proapoptotic proteins into the cytosol is the key event in apoptosis signaling, leading to the activation of caspases. Once in the cytosol, cytochrome c triggers the formation of a caspase-activating protein complex called the apoptosome, whereas Smac/Diablo and Omi/htra2 antagonize the caspase inhibitory effect of inhibitor of apoptosis proteins (IAPs). Here, we identify diarylurea compounds as effective inhibitors of the cytochrome c-induced formation of the active, approximately 700-kDa apoptosome complex and caspase activation. Using diarylureas to inhibit the formation of the apoptosome complex, we demonstrated that cytochrome c, rather than IAP antagonists, is the major mitochondrial caspase activation factor in tumor cells treated with tumor necrosis factor. Thus, we have identified a novel class of compounds that inhibits apoptosis by blocking the activation of the initiator caspase 9 by directly inhibiting the formation of the apoptosome complex. This mechanism of action is different from that employed by the widely used tetrapeptide inhibitors of caspases or known endogenous apoptosis inhibitors, such as Bcl-2 and IAPs. Thus, these compounds provide a novel specific tool to investigate the role of the apoptosome in mitochondrion-dependent death paradigms. PMID- 14560027 TI - Death receptor-induced activation of initiator caspase 8 is antagonized by serine/threonine kinase PAK4. AB - Normal cell growth requires a precisely controlled balance between cell death and survival. This involves activation of different types of intracellular signaling cascades within the cell. While some types of signaling proteins regulate apoptosis, or programmed cell death, other proteins within the cell can promote survival. The serine/threonine kinase PAK4 can protect cells from apoptosis in response to several different types of stimuli. As is the case for other members of the p21-activated kinase (PAK) family, one way that PAK4 may promote cell survival is by phosphorylating and thereby inhibiting the proapoptotic protein Bad. This leads in turn to the inhibition of effector caspases such as caspase 3. Here we show that in response to cytokines which activate death domain-containing receptors, such as the tumor necrosis factor and Fas receptors, PAK4 can inhibit the death signal by a different mechanism. Under these conditions, PAK4 inhibits apoptosis early in the caspase cascade, antagonizing the activation of initiator caspase 8. This inhibition, which does not require PAK4's kinase activity, may involve inhibition of caspase 8 recruitment to the death domain receptors. This role in regulating initiator caspases is an entirely novel role for the PAK proteins and suggests a new mechanism by which these proteins promote cell survival. PMID- 14560028 TI - Mutations in yeast replication proteins that increase CAG/CTG expansions also increase repeat fragility. AB - Expansion of trinucleotide repeats (TNRs) is the causative mutation in several human genetic diseases. Expanded TNR tracts are both unstable (changing in length) and fragile (displaying an increased propensity to break). We have investigated the relationship between fidelity of lagging-strand replication and both stability and fragility of TNRs. We devised a new yeast artificial chromomosme (YAC)-based assay for chromosome breakage to analyze fragility of CAG/CTG tracts in mutants deficient for proteins involved in lagging-strand replication: Fen1/Rad27, an endo/exonuclease involved in Okazaki fragment maturation, the nuclease/helicase Dna2, RNase HI, DNA ligase, polymerase delta, and primase. We found that deletion of RAD27 caused a large increase in breakage of short and long CAG/CTG tracts, and defects in DNA ligase and primase increased breakage of long tracts. We also found a correlation between mutations that increase CAG/CTG tract breakage and those that increase repeat expansion. These results suggest that processes that generate strand breaks, such as faulty Okazaki fragment processing or DNA repair, are an important source of TNR expansions. PMID- 14560029 TI - Swi1 prevents replication fork collapse and controls checkpoint kinase Cds1. AB - The replication checkpoint is a dedicated sensor-response system activated by impeded replication forks. It stabilizes stalled forks and arrests division, thereby preserving genome integrity and promoting cell survival. In budding yeast, Tof1 is thought to act as a specific mediator of the replication checkpoint signal that activates the effector kinase Rad53. Here we report studies of fission yeast Swi1, a Tof1-related protein required for a programmed fork-pausing event necessary for mating type switching. Our studies have shown that Swi1 is vital for proficient activation of the Rad53-like checkpoint kinase Cds1. Together they are required to prevent fork collapse in the ribosomal DNA repeats, and they also prevent irreversible fork arrest at a newly identified hydroxyurea pause site. Swi1 also has Cds1-independent functions. Rad22 DNA repair foci form during S phase in swi1 mutants and to a lesser extent in cds1 mutants, indicative of fork collapse. Mus81, a DNA endonuclease required for recovery from collapsed forks, is vital in swi1 but not cds1 mutants. Swi1 is recruited to chromatin during S phase. We propose that Swi1 stabilizes replication forks in a configuration that is recognized by replication checkpoint sensors. PMID- 14560030 TI - Molecular mechanism for a role of SHP2 in epidermal growth factor receptor signaling. AB - The Src homology 2-containing phosphotyrosine phosphatase (SHP2) is primarily a positive effector of receptor tyrosine kinase signaling. However, the molecular mechanism by which SHP2 effects its biological function is unknown. In this report, we provide evidence that defines the molecular mechanism and site of action of SHP2 in the epidermal growth factor-induced mitogenic pathway. We demonstrate that SHP2 acts upstream of Ras and functions by increasing the half life of activated Ras (GTP-Ras) in the cell by interfering with the process of Ras inactivation catalyzed by Ras GTPase-activating protein (RasGAP). It does so by inhibition of tyrosine phosphorylation-dependent translocation of RasGAP to the plasma membrane, to its substrate (GTP-Ras) microdomain. Inhibition is achieved through the dephosphorylation of RasGAP binding sites at the level of the plasma membrane. We have identified Tyr992 of the epidermal growth factor receptor (EGFR) to be one such site, since its mutation to Phe renders the EGFR refractory to the effect of dominant-negative SHP2. To our knowledge, this is the first report to outline the site and molecular mechanism of action of SHP2 in EGFR signaling, which may also serve as a model to describe its role in other receptor tyrosine kinase signaling pathways. PMID- 14560031 TI - In vivo evidence that defects in the transcriptional elongation factors RPB2, TFIIS, and SPT5 enhance upstream poly(A) site utilization. AB - While a number of proteins are involved in elongation processes, the mechanism for action of most of these factors remains unclear primarily because of the lack of suitable in vivo model systems. We identified in yeast several genes that contain internal poly(A) sites whose full-length mRNA formation is reduced by mutations in RNA polymerase II subunit RPB2, elongation factor SPT5, or TFIIS. RPB2 and SPT5 defects also promoted the utilization of upstream poly(A) sites for genes that contain multiple 3' poly(A) signaling sequences, supporting a role for elongation in differential poly(A) site choice. Our data suggest that elongation defects cause increased transcriptional pausing or arrest that results in increased utilization of internal or upstream poly(A) sites. Transcriptional pausing or arrest can therefore be visualized in vivo if a gene contains internal poly(A) sites, allowing biochemical and genetic study of the elongation process. PMID- 14560032 TI - Deficiency in the organic cation transporters 1 and 2 (Oct1/Oct2 [Slc22a1/Slc22a2]) in mice abolishes renal secretion of organic cations. AB - The polyspecific organic cation transporters 1 and 2 (Oct1 and -2) transport a broad range of substrates, including drugs, toxins, and endogenous compounds. Their strategic localization in the basolateral membrane of epithelial cells in the liver, intestine (Oct1), and kidney (Oct1 and Oct2) suggests that they play an essential role in removing noxious compounds from the body. We previously showed that in Oct1(-/-) mice, the hepatic uptake and intestinal excretion of organic cations are greatly reduced. Since Oct1 and Oct2 have extensively overlapping substrate specificities, they might be functionally redundant. To investigate the pharmacologic and physiologic roles of these proteins, we generated Oct2 single-knockout and Oct1/2 double-knockout mice. Oct2(-/-) and Oct1/2(-/-) mice are viable and fertile and display no obvious phenotypic abnormalities. Absence of Oct2 in itself had little effect on the pharmacokinetics of tetraethylammonium (TEA), but in Oct1/2(-/-) mice, renal secretion of this compound was completely abolished, leaving only glomerular filtration as a TEA clearance mechanism. As a consequence, levels of TEA were substantially increased in the plasma of Oct1/2(-/-) mice. This study shows that Oct1 and Oct2 together are essential for renal secretion of (small) organic cations. A deficiency in these proteins may thus result in increased drug sensitivity and toxicity. PMID- 14560033 TI - Trypanosoma brucei RNA editing complex: band II is structurally critical and maintains band V ligase, which is nonessential. AB - Maturation of Trypanosoma brucei mitochondrial mRNA involves massive posttranscriptional insertion and deletion of uridine residues. This RNA editing utilizes an enzymatic complex with seven major proteins, band I through band VII. We here use RNA interference (RNAi) to examine the band II and band V proteins. Band II is found essential for viability; it is needed to maintain the normal structure of the editing complex and to retain the band V ligase protein. Previously, band III was found essential for certain activities, including maintenance of the editing complex and retention of the band IV ligase protein. Thus, band II and band V form a protein pair with features analogous to the band III/band IV ligase pair. Since band V is specific for U insertion and since band IV is needed for U deletion, their parallel organization suggests that the editing complex has a pseudosymmetry. However, unlike the essential band IV ligase, RNAi to band V has only a morphological but no growth rate effect, suggesting that it is stimulatory but nonessential. Indeed, in vitro analysis of band V RNAi cell extract demonstrates that band IV can seal U insertion when band V is lacking. Thus, band IV ligase is the first activity of the basic editing complex shown able to serve in both forms of editing. Our studies also indicate that the U insertional portion may be less central in the editing complex than the corresponding U deletional portion. PMID- 14560034 TI - Agonist and chemopreventative ligands induce differential transcriptional cofactor recruitment by aryl hydrocarbon receptor. AB - Aryl hydrocarbon receptor (AHR) is a transcription factor whose activity is regulated by environmental agents, including several carcinogenic agonists. We measured recruitment of AHR and associated proteins to the human cytochrome P4501A1 gene promoter in vivo. Upon treatment with the agonist beta naphthoflavone, AHR is rapidly associated with the promoter and recruits the three members of the p160 family of coactivators as well as the p300 histone acetyltransferase, leading to recruitment of RNA polymerase II (Pol II) and induction of gene transcription. AHR, coactivators, and Pol II cycle on and off the promoter, with a period of approximately 60 min. In contrast, the chemopreventative AHR ligand 3,3'-diindolylmethane promotes AHR nuclear translocation and p160 coactivator recruitment but, remarkably, fails to recruit Pol II or cause histone acetylation. This novel mechanism of receptor antagonism may account for the antitumor properties of chemopreventative compounds targeting the AHR. PMID- 14560036 TI - Angiotensin subtype 1 rReceptor (AT1) blockade improves vasorelaxation in heart failure by up-regulation of endothelial nitric-oxide synthase via activation of the AT2 receptor. AB - To determine whether angiotensin receptor blockade decreases vascular tone in heart failure by improving endothelial-dependent vasorelaxation and increasing nitric oxide (NO) bioavailability, we treated infarcted adult male Sprague-Dawley rats with candesartan for 7 days or 8 weeks (10 mg/kg/day in drinking water). Candesartan, at both time points, lowered left ventricular (LV) systolic pressure (P < 0.05) (122 +/- 22 versus 74 +/- 16 and 73 +/- 10 mm Hg) and LV dP/dt (5914 +/- 1294 versus 2857 +/- 1672 versus 3175 +/- 769 mm Hg/s), but lowered LV end diastolic pressure only at 8 weeks (16.9 +/- 9.7 versus 11.2 +/- 5.7 versus 6.9 +/- 5.3 mm Hg). The vasorelaxation response to acetylcholine (ACh) in thoracic aortic segments was decreased with infarction (P < 0.05), remained unchanged with 1 week of candesartan, but increased 84 and 86% at 10-4 and 10-5 M ACh (P < 0.05) at 8 weeks. The enhanced candesartan-induced vasorelaxation at 8 weeks was abolished with NG-nitro-l-arginine methyl ester (200 microM). In bovine pulmonary endothelial cells, 20 microM candesartan increased endothelial nitric-oxide synthase (eNOS) protein levels (P < 0.05) (28.9 +/- 2.6 versus 16.1 +/- 3.7 intensity units/microg of protein); the increased eNOS was abolished by a specific angiotensin subtype 2 (AT2) receptor antagonist, PD 123319. These data suggest that AT1 receptor blockade enhances vasorelaxation in heart failure by increasing NO bioavailability, in part via an AT2 receptor-mediated up-regulation of eNOS protein. PMID- 14560035 TI - BARD1 participates with BRCA1 in homology-directed repair of chromosome breaks. AB - The BRCA1 tumor suppressor has been implicated in the maintenance of chromosomal stability through homology-directed repair of DNA double-strand breaks. Much of the BRCA1 in cells forms a heterodimeric complex with a structurally related protein BARD1. We report that expression of truncated mouse or human BARD1 peptides capable of interacting with Brca1 results in a homologous-repair deficiency. Repair is mildly reduced in Brca1 wild-type cells and severely reduced in cells that harbor a Brca1 splice product deleted for exon 11. Nuclear localization of the Brca1 or BARD1 peptides is not compromised, implying that the repair deficiency is caused by a more direct effect on repair. The tumor suppressor activity of BRCA1 may require the participation of BARD1 to maintain chromosome integrity through the homologous-repair pathway. PMID- 14560037 TI - Inhibition of adrenal cortical steroid formation by procaine is mediated by reduction of the cAMP-induced 3-hydroxy-3-methylglutaryl-coenzyme A reductase messenger ribonucleic acid levels. AB - Elevated glucocorticoid levels are associated with many diseases, including age related depression, hypertension, Alzheimer's disease, and acquired immunodeficiency syndrome. Cortisol-lowering agents could provide useful complementary therapy for these disorders. We examined the effect of procaine and procaine in a pharmaceutical formulation on adrenal cortical steroid formation. Procaine inhibited dibutyryl cyclic AMP (dbcAMP)-induced corticosteroid synthesis by murine Y1 and human H295R adrenal cells in a dose-dependent manner without affecting basal steroid formation. Treatment of rats with the procaine-based formulation reduced circulating corticosterone levels. This steroidogenesis inhibiting activity of procaine was not observed in Leydig cells, suggesting that the effect was specific to adrenocortical cells. In search of the mechanism underlying this inhibitory effect on cAMP-induced corticosteroidogenesis, procaine was found to affect neither the cAMP-dependent protein kinase activity nor key proteins involved in cholesterol transport into mitochondria, cytochrome P450 side chain cleavage enzyme expression, and enzymatic activities associated with cholesterol metabolism to final steroid products. However, procaine reduced in a dose-dependent manner the 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA) activity and the dbcAMP-induced HMG-CoA reductase mRNA levels by affecting mRNA stability. These data suggest that the inhibitory effect of procaine on cAMP-induced corticosteroid formation is due to the reduced synthesis of cholesterol. This modulatory effect of procaine on HMG-CoA reductase mRNA expression was also seen in dbcAMP-stimulated Hepa1-6 mouse liver hepatoma cells. Taken together, these results suggest that procaine may provide a pharmacological means for the control of hormone-induced HMG-CoA reductase mRNA expression and hypercortisolemia. PMID- 14560038 TI - Inhibition of platelet-activating factor (PAF) acetylhydrolase by methyl arachidonyl fluorophosphonate potentiates PAF synthesis in thrombin-stimulated human coronary artery endothelial cells. AB - We have previously demonstrated that thrombin stimulation of endothelial cells results in increased membrane-associated, Ca(2+)-independent phospholipase A2 (iPLA2) activity, accelerated hydrolysis of membrane plasmalogen phospholipids, and production of several biologically active phospholipid metabolites, including prostacyclin and platelet-activating factor (PAF) that is abolished by pretreatment with the iPLA2-selective inhibitor bromoenol lactone. This study was designed to further investigate the role of alternative PLA2 inhibitors, including methyl arachidonyl fluorophosphonate (MAFP, an inhibitor of cytosolic PLA2 isoforms), on phospholipid turnover and PAF production from thrombin stimulated human coronary artery endothelial cells (HCAECs). Paradoxically, pretreatment of HCAEC with MAFP (5-25 microM) resulted in a significant increase in PAF production in both unstimulated and thrombin-stimulated cells that was found to be a direct result of inhibition of PAF acetylhydrolase (PAF-AH) activity. Pretreatment with MAFP did not significantly inhibit HCAEC PLA2 activity, possibly due to the localization of PLA2 activity in the membrane fraction rather than the cytosol. Bromoenol lactone did not inhibit PAF-AH activity, even at concentrations as high as 20 microM. We conclude that MAFP augments thrombin-stimulated PAF production by inhibition of PAF catabolism without affecting membrane-associated iPLA2 activity. PMID- 14560039 TI - Peroxisome proliferator-activated receptor [gamma] agonist provides superior renal protection versus angiotensin-converting enzyme inhibition in a rat model of type 2 diabetes with obesity. AB - The inbred obese Zucker (ZDF/Gmi, fa/fa) rat develops severe hyperglycemia and also exhibits severe renal disease. In this study, we compared the relative benefits of long-term treatment with angiotensin-converting enzyme inhibition (ACEI) to a peroxisome proliferator-activated receptor gamma (PPARgamma) agonist. Four groups of obese inbred Zucker rats were studied over a 6-month observation period; untreated animals, rats treated with ACEI alone, rats treated with PPARgamma agonist alone, and rats treated with a combination of ACEI and PPARgamma agonist. PPARgamma agonist treatment normalized plasma glucose and led to massive increases in body weight. Both ACEI and PPARgamma agonist were effective in reducing the proteinuria and glomerular and tubular kidney damage. However, the PPARgamma agonist exerted superior renal protection compared with ACEI, in this model of spontaneously occurring chronic renal disease in the diabetic, obese inbred Zucker rat. Of note, although ACEI lowered blood pressure, there was no difference in glomerular blood pressure in any group at the end of the study. The glomerular filtration rate (GFR) was improved by ACEI with a borderline effect of PPARgamma agonist alone. A mild additive protection on GFR and tubulointerstitial damage was seen with the combination. Based on the literature it is likely that the superior protection by PPARgamma agonist versus glomerular and tubular damage as well as proteinuria extends beyond glycemic and lipidmic control and also reflects direct, protective intrarenal actions of the PPARgamma agonists. PMID- 14560040 TI - Subtype-selective up-regulation by chronic nicotine of high-affinity nicotinic receptors in rat brain demonstrated by receptor autoradiography. AB - Subtypes of neuronal nicotinic acetylcholine receptors (nAChRs) are differentially sensitive to up-regulation by chronic nicotine exposure in vitro. To determine whether this occurs in animals, rats were implanted with minipumps containing saline +/- nicotine (6.0 mg/kg/rat/day) for 14 days. Autoradiography with [125I]epibatidine using 3-(2(S)-azetidinylmethoxy)pyridine dihydrochloride (A-85380) or cytisine as selective competitors allowed quantitative measurement in 33 regions of 3 families of nAChR binding, with properties of alpha4beta2, alpha3beta4, and alpha3/alpha6beta2. Chronic nicotine exposure caused increases of 20 to 100% for alpha4beta2-like binding in most regions surveyed. However, binding to this subtype was not increased in some regions, including habenulopeduncular structures, certain thalamic nuclei, and several brainstem regions. In 9 of 33 regions, including catecholaminergic areas and visual structures, alpha3/alpha6beta2-like binding represented >10% of total binding. Binding to this subtype was up-regulated by nicotine in only two of these nine regions: the nucleus accumbens and superior colliculus. alpha3beta4-Like binding represented >10% of total in 15 of the 33 regions surveyed. Binding to this subtype was increased by nicotine in only 1 of these 15 regions, and actually decreased in subiculum and cerebellum. These studies yielded two principal findings. First, chronic nicotine exposure selectively up-regulates alpha4beta2 like binding, with relatively little effect on alpha3/alpha6beta2-like and alpha3beta4-like binding in vivo. Second, up-regulation by chronic nicotine exposure shows considerable regional variation. Differential subtype sensitivity to chronic nicotine exposure may contribute to altered pharmacological response in individuals who smoke or use nicotine replacement therapy. PMID- 14560042 TI - The thromboxane receptor antagonist PBT-3, a hepoxilin stable analog, selectively antagonizes the TPalpha isoform in transfected COS-7 cells. AB - The hepoxilin analog PBT-3 [10(S)-hydroxy-11,12-cyclopropyleicosa-5Z,8Z,14Z trienoic acid methyl ester] was previously shown to inhibit the aggregation of human platelets and to antagonize the binding of the thromboxane receptor agonist I-BOP [[1S-[1alpha,2alpha (Z),3beta(1E,3S*),4alpha]]-7-[3-[3-hydroxy-4-(4 iodophenoxy)-1-butenyl]-7-oxabicyclo[2.2.1]hept-2-yl]-5-heptenoic acid] in human platelets (Pace-Asciak et al., 2002). We show herein that PBT-3 inhibits, to different degrees, binding of the TP receptor antagonist [3H]SQ 29,548 [[1S [1alpha,2alpha (Z),3alpha,4alpha]]-7-[3-[[2 [(phenylamino)carbonyl]hydrazino]methyl]-7-oxabicyclo[2.2. 1]hept-2-yl]-5 heptenoic acid], to the TP receptor isoforms in TPalpha- and TPbeta-transfected COS-7 cells. These isoforms possess a different tail length, the alpha being shorter than the beta isoform. In contrast, SQ 29,548 shows no selection for the two TP isoforms. The IC50 value for PBT-3 = 2.0 +/- 0.3 x 10-7 M was observed for TPalpha, whereas this was one-sixth less active on the TPbeta isoform (IC50 = 1.2 +/- 0.2 x 10-6 M), suggesting selectivity for the TPalpha isoform. To investigate whether the tail contributes to the difference in competition binding by PBT-3, we investigated the tailless TP isoform expressed in transfected COS-7 cells. Its IC50 was similar to that of the TPalpha isoform. In additional studies, we investigated the effect of PBT-3 on the collagen and I-BOP evoked intracellular calcium release and on the collagen and I-BOP evoked phosphorylation of pleckstrin. PBT-3 blocked both pathways further demonstrating its TP receptor antagonist activity. These results demonstrate that the action of PBT-3 in inhibiting platelet aggregation is mediated via inhibition of the TPalpha isoform of the thromboxane receptor and that the tail may play an important role in recognition of this TP receptor antagonist. PMID- 14560041 TI - Methylnaltrexone antagonizes opioid-mediated enhancement of HIV infection of human blood mononuclear phagocytes. AB - Opioid abuse has been postulated as a cofactor in the immunopathogenesis of human immunodeficiency virus (HIV) infection and AIDS. We and others have recently demonstrated that opioid enhances HIV infection of human macrophages through modulation of beta-chemokines and the CCR5 receptor and that this effect is reversed by naltrexone, a tertiary opioid antagonist. Tertiary opioid antagonists cannot be used in opioid-dependent patients because they precipitate withdrawal or reversal of analgesia. We determined whether the quaternary opioid antagonist methylnaltrexone (MNTX), now in phase III clinical trials for opioid-induced constipation, reverses the opioid-mediated enhancement of HIV infection of macrophages at clinically relevant doses. MNTX completely abrogated morphine induced HIV Bal strain infection of macrophages. MNTX also inhibited the R5 strain (ADA) envelope-pseudotyped HIV replication induced by morphine. Furthermore, MNTX abolished morphine-mediated up-regulation of CCR5 receptor expression. The ability of MNTX to block opioid-induced CCR5 expression and HIV replication at clinically relevant doses may have additional benefit for opioid abusers with HIV infection, or patients with AIDS pain receiving opioids. PMID- 14560043 TI - Adenosine biosynthesis in the collecting duct. AB - Adenosine regulates tubular transport in collecting ducts (CDs); however, the sources of adenosine that modulate ion transport in CDs are unknown. The extracellular cAMP-adenosine pathway refers to the conversion of cAMP to AMP by ectophosphodiesterase, followed by metabolism of AMP to adenosine by ecto-5' nucleotidase, with all steps occurring in the extracellular compartment. The goal of this study was to assess whether the extracellular cAMP-adenosine pathway exists in CDs. Studies were conducted in both freshly isolated CDs and in CD cells in culture (first passage) that were derived from isolated CDs. Purity of CDs was confirmed by microscopy, by Western blotting for aquaporin-1, aquaporin 2, bumetanide-sensitive cotransporter type 1, and thiazide-sensitive cotransporter; and by reverse transcription-polymerase chain reaction for adenosine receptors. Both freshly isolated CDs and CD cells in culture converted exogenous cAMP to AMP and adenosine. In both freshly isolated CDs and CD cells in culture, conversion of cAMP to AMP and adenosine was affected by a broad-spectrum phosphodiesterase inhibitor (3-isobutyl-1-methylxanthine), an ectophosphodiesterase inhibitor (1,3-dipropyl-8-p-sulfophenylxanthine), and a blocker of ecto-5'-nucleotidase (alpha,beta-methylene-adenosine-5'-diphosphate) in a manner consistent with exogenous cAMP being processed by the extracellular cAMP-adenosine pathway. In CD cells in culture, stimulation of adenylyl cyclase increased extracellular concentrations of cAMP, AMP, and adenosine, and these changes were also modulated by the aforementioned inhibitors in a manner consistent with the extracellular cAMP-adenosine pathway. In conclusion, the extracellular cAMP-adenosine pathway is an important source of adenosine in CDs. PMID- 14560044 TI - Gallium arsenide selectively up-regulates inflammatory cytokine expression at exposure site. AB - Gallium arsenide (GaAs), a technologically and economically important semiconductor, is widely utilized in both military and commercial applications. This chemical is a potential health hazard as a carcinogen and immunotoxicant. We previously reported that macrophages at the exposure site exhibit characteristics of activation. In vitro culture of macrophages with GaAs fails to recapitulate the in vivo phenotype, suggesting that complete GaAs-mediated activation in vivo may require other cells or components found in the body's microenvironment. Our present study examined the role of cytokines upon GaAs-mediated macrophage activation. Intraperitoneal administration of GaAs elicited rapid specific recruitment of blood monocytes to the exposure site. This recruitment occurred concomitant with up-regulation of 17 chemokine and inflammatory cytokine mRNAs, while transcripts of three inhibitory cytokines diminished. Administration of latex beads caused less cytokine induction than GaAs, indicating that changes in mRNA levels could not be attributed to phagocytosis. Four representative chemokines and cytokines were selected for further analysis. Increased cytokine mRNA expression was paralleled by similar increases in cytokine protein levels, and secreted protein products were detected in peritoneal fluid. Cytokine protein expression was constrained to myeloid cells, and to a lesser extent to B cells. Alterations in patterns of cytokine gene expression elucidate mechanisms for increased cellular activation and antigen processing, and modulation of the inflammatory response. Our findings indicate that in vivo GaAs exposure alters cytokine gene expression, which may lead to an inflammatory reaction and contribute to pathological tissue damage. PMID- 14560045 TI - Protein translocation in photoreceptor light adaptation: a common theme in vertebrate and invertebrate vision. AB - How do our eyes adjust to daily light levels that vary by almost 11 orders of magnitude? Research shows that, in both vertebrates and invertebrates, signaling proteins are translocated in a light-dependent manner between the photoreceptor cellular compartments where visual transduction takes place, and the rest of the photoreceptor cell. Protein translocation is likely to contribute to photoreceptor light adaptation by adjusting the sensitivity and speed of photoresponse to ever-changing conditions of ambient illumination. PMID- 14560046 TI - PDE4: arrested at the border. AB - Compartmentation plays a critical role in determining the specificity and efficacy of signaling by adenosine 3',5'-monophosphate (cAMP) and other second messengers. It has become apparent that all of the protein components of the cAMP signaling pathway are subject to regulation and may be localized to particular subcellular compartments. This Perspective discusses recent research concerning the importance of cAMP phosphodiesterase localization in cAMP signaling, functional implications of the organization of this pathway's protein components into macromolecular complexes, and common themes in the compartmentation of cAMP- and Ca2+-mediated signaling pathways. PMID- 14560048 TI - Vicinal disulfide turns. AB - The formation of a disulfide bond between adjacent cysteine residues is accompanied by the formation of a tight turn of the protein backbone. In nearly 90% of the structures analyzed a type VIII turn was found. The peptide bond between the two cysteines is in a distorted trans conformation, the omega torsion angle ranges from 159 to -133 degrees, with an average value of 171 degrees. The constrained nature of the vicinal disulfide turn and the pronounced difference observed between the oxidized and reduced states, suggests that vicinal disulfides may be employed as a 'redox-activated' conformational switch. PMID- 14560047 TI - Coupling the iron-responsive element to GFP--an inducible system to study translation in a single living cell. AB - Local protein synthesis in a cell represents an elegant mechanism to achieve important biological phenomena such as cell migration, body axis formation during embryonic development and establishment of cell polarity. A prerequisite to studying translation in a restricted cellular compartment is the ability to unambiguously discriminate between proteins that arise through local protein synthesis and those that reach the site of interest by diffusion or transport. To tackle this problem, we set up a green fluorescent protein (GFP)-based reporter system that allows one to uncouple the translation of reporter gene mRNA from its subcellular localization. The system is based on the iron-responsive element, which regulates the translation of both endogenous ferritin and transferrin transcripts in response to changes in iron concentration. Translation of the reporter messenger RNA (mRNA) is thus dependent on iron in the medium; both its transcription and localization, however, are unaffected. Known targeting sequences can be used to direct the mRNA transcript to a subcellular compartment of interest. For instance, the full-length 3'-untranslated region of calcium/calmodulin-dependent protein kinase IIalpha mRNA can be added to the construct, after the stop codon of the GFP sequence, to selectively target the transcript into the dendrites of transiently transfected hippocampal neurons. This novel fluorescent assay will allow us to address a number of important biological questions in living mammalian cells. PMID- 14560049 TI - PROSPECT II: protein structure prediction program for genome-scale applications. AB - A new method for fold recognition is developed and added to the general protein structure prediction package PROSPECT (http://compbio.ornl.gov/PROSPECT/). The new method (PROSPECT II) has four key features. (i) We have developed an efficient way to utilize the evolutionary information for evaluating the threading potentials including singleton and pairwise energies. (ii) We have developed a two-stage threading strategy: (a) threading using dynamic programming without considering the pairwise energy and (b) fold recognition considering all the energy terms, including the pairwise energy calculated from the dynamic programming threading alignments. (iii) We have developed a combined z-score scheme for fold recognition, which takes into consideration the z-scores of each energy term. (iv) Based on the z-scores, we have developed a confidence index, which measures the reliability of a prediction and a possible structure-function relationship based on a statistical analysis of a large data set consisting of threadings of 600 query proteins against the entire FSSP templates. Tests on several benchmark sets indicate that the evolutionary information and other new features of PROSPECT II greatly improve the alignment accuracy. We also demonstrate that the performance of PROSPECT II on fold recognition is significantly better than any other method available at all levels of similarity. Improvement in the sensitivity of the fold recognition, especially at the superfamily and fold levels, makes PROSPECT II a reliable and fully automated protein structure and function prediction program for genome-scale applications. PMID- 14560050 TI - Prediction of protein secondary structure with a reliability score estimated by local sequence clustering. AB - Most algorithms for protein secondary structure prediction are based on machine learning techniques, e.g. neural networks. Good architectures and learning methods have improved the performance continuously. The introduction of profile methods, e.g. PSI-BLAST, has been a major breakthrough in increasing the prediction accuracy to close to 80%. In this paper, a brute-force algorithm is proposed and the reliability of each prediction is estimated by a z-score based on local sequence clustering. This algorithm is intended to perform well for those secondary structures in a protein whose formation is mainly dominated by the neighboring sequences and short-range interactions. A reliability z-score has been defined to estimate the goodness of a putative cluster found for a query sequence in a database. The database for prediction was constructed by experimentally determined, non-redundant protein structures with <25% sequence homology, a list maintained by PDBSELECT. Our test results have shown that this new algorithm, belonging to what is known as nearest neighbor methods, performed very well within the expectation of previous methods and that the reliability z score as defined was correlated with the reliability of prediction. This led to the possibility of making very accurate predictions for a few selected residues in a protein with an accuracy measure of Q3 > 80%. The further development of this algorithm, and a nucleation mechanism for protein folding are suggested. PMID- 14560051 TI - A new computational approach for real protein folding prediction. AB - An effective and fast minimization approach is proposed for the prediction of protein folding, in which the 'relative entropy' is used as a minimization function and the off-lattice model is used. In this approach, we only use the information of distances between the consecutive Calpha atoms along the peptide chain and a generalized form of the contact potential for 20 types of amino acids. Tests of the algorithm are performed on the real proteins. The root mean square deviations of the structures of eight folded target proteins versus the native structures are in a reasonable range. In principle, this method is an improvement on the energy minimization approach. PMID- 14560052 TI - How can free energy component analysis explain the difference in protein stability caused by amino acid substitutions? Effect of three hydrophobic mutations at the 56th residue on the stability of human lysozyme. AB - To elucidate the molecular mechanism of thermal stability, it is essential to determine what are the major free energy components that contribute significantly to the total free energy difference caused by amino acid mutations. In this work, we carried out free energy calculations based on all-atom molecular dynamics simulations to investigate the effect of three hydrophobic mutations at the same position, I56A, I56V and I56F of human lysozyme. The calculated free energy differences are in good agreement with the experimental values in all cases. From free energy component analysis, we found that small changes in stability in the I56A and I56V mutants originate from the short-range Lennard-Jones interactions, whereas the I56F mutant is largely destabilized owing to the changes in the long range electrostatic interactions. The calculated results are also compared with the free energy components determined by an empirical relationship based on the native-state structure and thermodynamic data. Although this relationship has been shown to be very successful in reproducing the stability changes caused by various amino acid substitutions in several proteins, the changes of stability in I56V and I56F mutants are not reproduced very well. By comparing the free energy components calculated by these two approaches, we showed that the effect of the long-range interaction on the stability changes may be underestimated in the empirical relationships when the structural change caused by mutation is relatively small, as in I56F. It is also suggested that estimation of the change in accessible surface area, deltadeltaASA, may be overestimated if the structure around the mutation site in the denatured state is native-like, which would cause overestimation of the free energy change as in the case of I56V. Our results clearly show that the combined approach of the free energy calculation based on the all-atom molecular dynamics simulation and the empirical relationships is very useful for understanding the detailed mechanism of protein stability. PMID- 14560053 TI - Structure-oriented rational design of chymotrypsin inhibitor models. AB - Three peptides modelling a highly potent, 35-residue chymotrypsin inhibitor (Schistocerca gregaria chymotrypsin inhibitor) were designed and synthesized by convergent peptide synthesis. For each model peptide, the inhibitory constant (Ki) on chymotrypsin and the solution structure were determined. In addition, molecular dynamics calculations were performed for all of them. Two models containing approximately half of the parent inhibitor (17 of 35 residues) were designed and subsequently found to have no substantial inhibitory activity (Ki values in the mM range). The third model composed of 24 amino acid residues proved to be an effective (Ki approximately 10(-7)) inhibitor of bovine chymotrypsin. Both the solution structure properties determined by NMR spectroscopy and the dynamic behaviour of the latter model system are comparable to the native inhibitor. In contrast, the structure and dynamics of the first two related model peptides show characteristic differences. We suggest that the conformation and flexibility of the modelled protease inhibitor are crucial for its biological efficiency. Moreover, the structural and dynamic features of the binding loop (28-33) and those of the rest of the molecule appear to be interdependent. Most importantly, these structural characteristics can be rationally modified, at least partially, by peptide design. PMID- 14560054 TI - Directed evolution of Thermotoga neapolitana xylose isomerase: high activity on glucose at low temperature and low pH. AB - The Thermotoga neapolitana xylose isomerase (TNXI) is extremely thermostable and optimally active at 95 degrees C. Its derivative, TNXI Val185Thr (V185T), is the most active type II xylose isomerase reported, with a catalytic efficiency of 25.1 s(-1) mM(-1) toward glucose at 80 degrees C (pH 7.0). To further optimize TNXI's potential industrial utility, two rounds of random mutagenesis and low temperature/low pH activity screening were performed using the TNXI V185T encoding gene as the template. Two highly active mutants were obtained, 3A2 (V185T/L282P) and 1F1 (V185T/L282P/F186S). 1F1 was more active than 3A2, which in turn was more active than TNXI V185T at all temperatures and pH values tested. 3A2 and 1F1's high activities at low temperatures were due to significantly lower activation energies (57 and 44 kJ/mol, respectively) than that of TNXI and V185T (87 kJ/mol). Mutation L282P introduced a kink in helix alpha7 of 3A2's (alpha/beta)8 barrel. Surprisingly, this mutation kinetically destabilized 3A2 only at pH 5.5. 1F1 displayed kinetic stability slightly above that of TNXI V185T. In 1F1, mutation F186S creates a cavity that disrupts a four-residue network of aromatic interactions. How the conformation of the neighboring residues is affected by this cavity and how these conformational changes increase 1F1's stability still remain unclear. PMID- 14560055 TI - Inhibition of the CD28-CD80 co-stimulation signal by a CD28-binding affibody ligand developed by combinatorial protein engineering. AB - CD28 is one of the key molecules for co-stimulatory signalling in T cells. Here, novel ligands (affibodies) showing selective binding to human CD28 (hCD28) have been selected by phage display technology from a protein library constructed through combinatorial mutagenesis of a 58-residue three-helix bundle domain derived from staphylococcal protein A. Analysis of selected affibodies showed a marked sequence homology and biosensor analyses showed that all investigated affibodies bound to hCD28 with micromolar affinities (KD). No cross-reactivity towards the related protein human CTLA-4 could be observed. This lack of cross reactivity to hCTLA-4 suggests that the recognition site on hCD28 for the affibodies resides outside the conserved MYPPPYY motif. The apparent binding affinity for hCD28 could be improved through fusion to an Fc fragment fusion partner, resulting in a divalent presentation of the affibody ligand. For the majority of selected anti-CD28 affibodies, in co-culture experiments involving Jurkat T-cells and CHO cell lines transfected to express human CD80 (hCD80) or LFA-3 (hLFA-3) on the cell surface, respectively, pre-incubation of Jurkat cells with affibodies resulted in inhibition of IL-2 production when they were co cultured with CHO (hCD80+) cells, but not with CHO (hLFA-3+) cells. For one affibody variant denoted Z(CD28:5) a clear concentration-dependent inhibition was seen, indicating that this affibody binds hCD28 and specifically interferes in the interaction between hCD28 and hCD80. PMID- 14560056 TI - Synthesis of a novel histidine analogue and its efficient incorporation into a protein in vivo. AB - Proteins containing unnatural amino acids have immense potential in biotechnology and medicine. We prepared several histidine analogues including a novel histidine analogue, beta-(1,2,3-triazol-4-yl)-DL-alanine. These histidine analogues were assayed for translational activity in histidine-auxotrophic Escherichia coli strain UTH780. We observed that several histidine analogues, including our novel histidine analogue, were efficiently incorporated into the protein in vivo; however, other analogues were rejected. These results suggest that the hydrogen atom at a specific position seriously affects incorporation. PMID- 14560057 TI - Stable, soluble T-cell receptor molecules for crystallization and therapeutics. AB - Antibody and T-cell receptors (TCRs) are the primary recognition molecules of the adaptive immune system. Antibodies have been extensively characterized and are being developed for a large number of therapeutic applications. This has been possible because of the ability to manufacture stable, soluble, monoclonal antibodies which retain the antigen specificity of B cells. Unlike antibodies, TCRs are not expressed in a soluble form, but are anchored to the T-cell surface by an insoluble trans-membrane domain. Characterization and development of TCRs has been hampered by the lack of suitable methods for producing them as soluble and stable proteins. Here we report the engineering of soluble human TCRs suitable for crystallization studies and potentially for in vivo therapeutic use. PMID- 14560058 TI - A method for estimating duration of illness in Alzheimer's disease. AB - BACKGROUND: We developed a set of questions for generating an estimate regarding the date of first symptoms to the nearest half-year. Physicians then revised this estimate in conjunction with medical record review and patient/informant interviews, and by testing the estimate by recall of life events. One experienced examiner rated 36 patients, and each was independently rated by a second, less experienced rater. The physician ratings were compared to each other and to an unstructured caregiver estimate of duration using Lin concordance coefficients. There was excellent agreement between independent physician raters (rho = 0.95, p < 0.001). Caregiver estimates of duration were usually shorter because of failure to relate the first symptoms to the onset of disease. PMID- 14560059 TI - Donepezil for the treatment of mild to moderate Alzheimer's disease in France: the economic implications. AB - In the present study, the socioeconomic impact of the use of the acetylcholinesterase inhibitor donepezil in patients with mild to moderate Alzheimer's disease (AD) living in France was examined. A model was created to extrapolate over a 3-year period the results from placebo-controlled trials together with epidemiological and prevalence data. Costs considered in the model were net societal costs associated with paid and unpaid assistance, general medical consumption and institutional care. The model suggested that delays in cognitive decline and functional dependence due to treatment reduced the time spent in institutional care and the burden on caregivers. Over a 3-year period, total net costs of caring for untreated patients with an initial Mini-Mental State Examination score ranging from 10 to 26 were EUR 53,206 compared with EUR 42,720 for a patient treated with donepezil--an annual cost saving of approximately EUR 3,500 per patient. Cost savings were mainly due to savings in unpaid caregiver time, which, apart from patient institutionalization, represented the most costly component of total care in this study but had no direct budgetary impact. Overall, these data suggest that donepezil is a cost effective treatment for mild to moderately impaired AD patients living in France. PMID- 14560061 TI - Rapidly progressive aphasic dementia with motor neuron disease: a distinctive clinical entity. AB - The association of motor neuron disease (MND) with rapidly progressive aphasic dementia has been recognized as a distinct clinical syndrome within the group of frontotemporal dementias (FTDs). Although the clinical and neuropsychological features of this syndrome have been defined, a small number of post-mortem studies have been published with heterogeneous neuropathological findings. We performed cognitive, neuro-imaging and neuropathological studies on a 71-year-old male with rapidly progressive aphasic dementia and MND. We initially found a selective non-fluent aphasia associated with hypoperfusion of the left frontotemporal cortex. Proton magnetic resonance spectroscopy revealed an asymmetric change of brain metabolites, with greater changes in the left temporal lobe. The bulbar manifestations of MND occurred over the following 6 months, and the patient died of bronchopneumonia. The neuropathological examination revealed loss of neurons in the hypoglossal nucleus and anterior horns of the cervical spinal cord with microvacuolation and dot-like ubiquitin-positive deposits in the frontoparietotemporal cortex, but no changes suggestive of Alzheimer's, Pick's or Lewy body disease. These findings support the conclusion that MND with rapidly progressive aphasic dementia is a distinctive clinical entity within the group of FTD-MND. PMID- 14560060 TI - Agitation in Alzheimer's disease is a manifestation of frontal lobe dysfunction. AB - OBJECTIVES: (1) To investigate the prevalence and characteristics of agitation in patients with Alzheimer's disease (AD) and other forms of dementia; (2) to explore the association between agitation and other clinical variables, including disease severity, functional impairment and other neuropsychiatric symptoms, and (3) to determine the predictors of agitation. METHODS: Data for 427 men and women with dementia from outpatient clinics of the University of California, Los Angeles Alzheimer's Disease Center were analyzed. There were 277 patients with AD, 43 with vascular dementia, 47 with mixed dementia, 45 with frontotemporal dementia and 15 with dementia with Lewy bodies. Patients were evaluated with the Mini-Mental State Examination (MMSE), Neuropsychiatric Inventory (NPI), Functional Activities Questionnaire (FAQ), neuropsychological tests and the Caregiver Appraisal instrument. SPSS10 was utilized for statistical analysis. RESULTS: There was no difference in agitation subscale scores among patients with dementia of various etiologies. In patients with AD, there was increased prevalence of agitation with increasing dementia severity. Agitation contributed substantially to caregiver burden and impact. There was a significant correlation between the FAQ and the NPI agitation subscale score after adjusting for MMSE scores. Delusion, disinhibition and irritability subscale scores in AD patients were correlated with agitation across disease severity. Subscale scores of frontally mediated behaviors including irritability, delusions and disinhibition predicted most of the variance in agitation levels. CONCLUSION: Agitation is common in AD and other dementias and has a marked impact on caregivers. It is related to dementia severity and to specific types of associated psychopathology implicating frontal lobe dysfunction. The present study is the largest and most comprehensive assessment of agitation reported. The data suggest that agitation in AD is a frontal lobe syndrome. Frontal lobe dysfunction may predispose AD patients to agitation by exaggerating behavioral responses to many types of coexisting psychopathology or environmental provocations. PMID- 14560063 TI - Different classification systems yield different dementia occurrence among nonagenarians and centenarians. AB - Literature data consistently show different prevalence estimates of dementia when different classification systems are used in the same population. Very few data are available for the oldest old of the elderly. We investigated the occurrence of dementia among 34 nonagenarians and centenarians according to four classification systems: the American Psychiatric Association's Diagnostic and Statistical Manual of Mental Disorders, third edition revised (DSM-III-R) and fourth edition (DSM-IV), the World Health Organization's International Classification of Diseases, 10th revision (ICD-10), and the Cambridge Examination for Mental Disorders of the Elderly (CAMDEX). Cognitive functioning, work, social function and independence in activities of daily living were evaluated by using an extensive neuropsychological examination. The prevalence (95% CI) of dementia was the following: 47.1% (95% CI 30.3-63.8) with the DSM-III-R criteria, 41.2% (95% CI 24.6-57.7) with the DSM-IV criteria, 29.4% (95% CI 14.1-44.7) with the ICD-10 criteria and 38.2% (95% CI 21.9-54.6) with the CAMDEX. The factors that best predicted disagreement between DSM-III-R and DSM-IV were calculation impairment and the presence or absence of personality changes. DSM-III-R and ICD 10 were differentiated by the weight given to executive functions that all have to be impaired according to ICD-10, whereas progressive deterioration differentiated CAMDEX from DSM-III-R. It should be noted that although the DSM III-R diagnoses differ by a factor of 1.6 times from the ICD-10 and 1.2 times from the CAMDEX diagnoses, we are speaking about dementia, which is very frequent in nonagenarians and centenarians. Moreover, with regard to public health, an estimation of the number of subjects who will lose their autonomy is rather more useful and informative than simple prevalence figures of dementia by itself. In this light, classification systems, such as the ICD-10, that do not include impairment of social function as a criterion for assessing dementia become less adequate. PMID- 14560062 TI - Management of patients with Alzheimer's disease plus cerebrovascular disease: 12 month treatment with galantamine. AB - We evaluated the long-term cognitive effects and safety of galantamine 24 mg/day in patients with Alzheimer's disease plus cerebrovascular disease (AD + CVD or mixed dementia). Subgroup analysis was performed of patients with AD + CVD who participated in a 6-month, multicenter, randomized, double-blind, parallel-group study and a 6-month, open-label, active-treatment extension. METHOD: Two hundred and eighty-five patients with AD + CVD were randomized to receive either placebo (n = 97) or galantamine 24 mg/day (n = 188) for 6 months. Two hundred and thirty eight (84%) patients continued with the open-label phase of the study (86 from the placebo group, 152 from the galantamine group) and were treated with galantamine 24 mg/day. The primary efficacy measure was cognitive performance as assessed using the eleven-item cognitive subscale of the Alzheimer's Disease Assessment Scale (ADAS-cog/11). Standard safety evaluations and adverse-event monitoring were performed throughout the 12-month study period. Patients with AD + CVD treated with galantamine experienced statistically and clinically significant improvement in cognition at month 6 (mean change in ADAS-cog/11 score -1.1; p < or = 0.05 vs. baseline) and maintained their cognitive function for the entire 12-month study (mean change in ADAS-cog/11 score +0.1). In contrast, the cognitive function deteriorated among those in the placebo group (mean change in ADAS-cog/11 at month 6 +2.0; p < or = 0.001 vs. baseline). Patients with AD + CVD who were switched from placebo to galantamine for the open-label phase of the trial did show improvement in cognitive function; however, they never attained the same cognitive level as patients who had been treated with galantamine for the entire 12 months [mean (+/- SE) ADAS-cog/11 scores in the placebo/galantamine group 25.7 +/- 1.32 and 24.2 +/- 1.57 at months 6 and 12, respectively, and in the galantamine/galantamine group 21.5 +/- 0.87 and 22.2 +/- 1.06 at months 6 and 12, respectively]. The results of this subgroup analysis indicate that galantamine is effective for long-term maintenance of the cognitive function in patients with AD + CVD and is safe and well tolerated in this patient population. PMID- 14560064 TI - Detecting dementia: novel neuropsychological markers of preclinical Alzheimer's disease. AB - The results of a previous study have suggested that impaired performance on one neuropsychological test, CANTAB Paired Associates Learning (PAL), may serve as a marker for preclinical Alzheimer's disease (AD). In a group of individuals with 'questionable dementia', the baseline PAL performance was found to correlate significantly with subsequent deterioration in global cognitive function over an 8-month period. The present paper reports diagnostic outcome data for the same individuals 32 months after the first assessment and evaluates the predictive diagnostic utility of baseline neuropsychological measures. Thirty-two months after joining the study, 11 of the 43 'questionable dementia' patients met the criteria for probable AD diagnosis ('converters') and 29 remained free from AD ('non-converters'). Logistic regression analysis revealed that two tests of memory, in combination, could be used to predict a later diagnosis of probable AD with a high level of accuracy [chi(2)(3) = 47.054, p < 0.0001]. As predicted, these tests are measures of visuospatial learning (CANTAB PAL) and, also, semantic memory (Graded Naming Test). These two tests in combination appear to be highly accurate in detecting cognitive dysfunction characteristic of preclinical AD. Using these tests, a simple algorithm is described for calculating, with 100% accuracy for this sample of 40 patients, the probability that an individual with mild memory impairments will go on to receive a diagnosis of probable AD. PMID- 14560065 TI - Screening for cognitive impairment in patients with acute stroke. AB - BACKGROUND: Formal assessment of cognitive impairment is rare in acute stroke, yet. It was the goal to utilize easy-to-apply established screening tests to assess cognitive impairment from any cause in patients with acute ischemic stroke. METHODS: 209 consecutive patients (69.8 +/- 13.3 years, mean +/- SD; 117 male, 92 female) admitted to an acute stroke unit, which serves as a community stroke center, and diagnosed as having acute cerebral ischemia from any cause were investigated within 24 h of stroke onset and in part followed up after 3 months. Orientation and aphasia were assessed with the ADAScog subscales orientation and aphasia, verbal memory with Buschke's Memory Impairment Screen, and concentration/working memory with a letter sorting test. RESULTS: On admission, 74.6% were impaired on the Memory Impairment Screen, 77.0% on the letter sorting test, 45.0 and 24.9% on the ADAScog subscales aphasia and orientation. Results of the Memory Impairment Screen and letter sorting test were similar at the follow-up after 3 months. CONCLUSION: Formal brief assessment of cognitive deficits with tests not requiring motor capabilities in unselected patients with an acute cerebral ischemic event reveals widespread deficits. This needs to be considered when obtaining informed consent from the patients and instructing them. Routine formalized screening may thus improve care for patients with acute stroke. Patients with impairment on admission should be followed up, diagnosed concerning preexisting or poststroke dementia with a more extensive workup and eventually treated. PMID- 14560066 TI - Economic considerations for health insurance coverage of emerging genetic tests. AB - Public and private health insurance plans face the question of whether to cover emerging genetic tests for cancer and other diseases. This paper outlines issues in the economic evaluation of new genetic tests, illustrating key methodological issues and policy implications with findings from a comprehensive and systematic review of the 14 full economic evaluations published over the past 5 years that have addressed both the costs and consequences of molecular genetic tests. Key questions for framing an evaluation include: whose viewpoint matters, which costs and consequences are relevant, and to which clinical alternatives should new genetic tests be compared? While economic evaluation research can inform coverage decisions about genetic tests, the coverage decision-making process must also inform economic researchers about the aims, context, and value systems within which genetic tests will be covered and practised. PMID- 14560067 TI - Reproductive risk factors related to socioeconomic status in pregnant women in southern Brazil. AB - To evaluate patterns of drug use during pregnancy and other potential reproductive risks in pregnant women, we applied a standard questionnaire to 412 pregnant women classified as low socioeconomic status (SES) or as middle/high SES, in two cities of South Brazil. 77% of the women used at least one medication during pregnancy. We observed significant differences, when comparing low SES and middle/high SES groups, for the following variables: teenage pregnancies (28.4 vs. 12.4%); self-medication (21.8 vs. 13.1%); smoking habits (21.5 vs. 5.1%); unplanned pregnancies (69.5 vs. 51.8%); unwanted pregnancies (31.3 vs. 10.9%), and abortion attempts (13.1 vs. 5.8%). The average number of drugs consumed, however, was not different between low SES and middle/high SES. We believe that better education and conditions for family planning are key points to improve this scenario. PMID- 14560068 TI - Validation of the dutch short form of the state scale of the Spielberger State Trait Anxiety Inventory: considerations for usage in screening outcomes. AB - OBJECTIVE: To validate the six-item short form of the state scale of the Spielberger State-Trait Anxiety Inventory (STAI) for usage in screening outcomes in a Dutch population receiving preconception counseling. METHODS: Men and women completed the 20-item full form of the STAI before (n = 310) and after preconception counseling (n = 147). Prorated scores of the six-item form were compared to the full form using Pearson's correlation coefficients and paired t tests. RESULTS: Cronbach's alpha for the Dutch translation of the short-form of the STAI-state was 0.83. The short form highly correlates with the full form (r = 0.95). The short form was able to discriminate between different levels of anxiety and was sensitive to change. CONCLUSION: The Dutch translation of the short form of the STAI-state has good reliability and validity and was found to be useful as a quick tool to evaluate the effectiveness of screening programs on anxiety levels. We believe our results will be applicable to other populations, although this needs to be confirmed in other studies. PMID- 14560069 TI - Between acculturation and ambivalence: knowledge of genetics and attitudes towards genetic testing in a consanguineous bedouin community. AB - The Bedouins of the Negev (Southern part of Israel) are a community at increased risk for genetic diseases and congenital anomalies as a result of frequent consanguinity (particularly patrilateral parallel-cousin marriage) and underutilization of prenatal genetic tests due to a Muslim ban on abortion. OBJECTIVE: To assess the knowledge and attitudes of Bedouin schoolchildren and their teachers towards a community-based, premarital carrier-matching program aimed at reducing the prevalence at birth of genetic diseases. METHODS: A questionnaire was presented to 61 teachers and 40 schoolchildren as part of guided interaction in small groups, conducted in Bedouin schools between 1999 and 2001. RESULTS: Susceptibility as well as knowledge of genetics were found to correlate with a positive attitude towards the genetics program among both teachers and pupils. However, pupils had a lower knowledge index as compared to teachers, and their attitudes were slightly less positive. CONCLUSION: The difference between teachers and pupils is discussed in the context of the latter's acculturation, which contradicts tradition and parental authority and can generate ambivalence. Attitudes are further discussed in the context of the Health Belief Model and the complex interplay of tradition, Islam, cousin marriage and biomedicine. PMID- 14560070 TI - Community diagnosis of maternal exposure to risk factors for congenital defects. AB - OBJECTIVES: This paper describes a community diagnosis of maternal exposure to risk factors for congenital defects in the population of Pelotas, RS, Brazil (400000 inhabitants). The authors investigated biological and demographic factors (maternal age, alcohol ingestion and smoking), social and economic factors (family income and type of work), and welfare factors (prenatal care, illnesses during pregnancy, drug therapy, and vaccinations). These factors were matched up with the Decalogue for Prevention of Congenital Defects recommended by the Latin American Collaborative Study of Congenital Malformations. METHODS: Data were collected in a population-based study in which mothers were interviewed in the hospitals of Pelotas, RS, during 1995. On each day, the first 3 women to give birth were interviewed in every city hospital. The sample resulted in a total of 3219 interviewed women. RESULTS AND CONCLUSIONS: Low-income groups are most exposed. They present the following potential risk factors: (a). outside the ideal age range for pregnancy; (b). fewer prenatal care appointments; (c). no prevention against rubella through vaccinations, and (d). more cigarette smoking. In the high-income groups, the potential risk factors were: (a). drug therapy (more common) and (b). more women working outside the home. We did not find any difference among social classes concerning: (a). illnesses during pregnancy and (b). ingestion of alcohol. When a general risk score was used, this figure was higher in the group of women who were not within ideal age for pregnancy. However, there was no difference among social classes. This has led us to conclude that delayed childbearing is a global risk whereas each social class has its own risks. PMID- 14560071 TI - Predictive genetic testing for breast cancer and Huntington's disease: the opinions of midwives and nurses in Flanders. AB - OBJECTIVE: Investigate attitudes of midwives and nurses regarding predictive testing for hereditary breast cancer and Huntington's disease. METHODS: Mail questionnaires were sent to 119 midwives and 881 nurses. RESULTS: The response rate was 58% for midwives, but only 33% for nurses. The attitude toward predictive testing in an asymptomatic adult was more favourable in case of hereditary breast cancer than in case of Huntington's disease, mostly because prevention or therapy is unavailable for the latter condition. Prevention of breast cancer through prophylactic mastectomy elicited adverse reactions among 40% of the participants. Predictive testing for these diseases in an adolescent at the own request was also rated favourably while testing in children at the parents' request was less acceptable. For both diseases, concealing carrier status information from the employer or the insurance company was judged as acceptable by the majority. CONCLUSION: Our findings contribute to the understanding of nurses' and especially midwives' attitudes to predictive testing, which may in turn serve as a basis for genetic education for these groups of health care professionals. PMID- 14560072 TI - Individualized pharmacogenetic therapy: a critical analysis. AB - OBJECTIVE: Individualized, or personalized, therapy is highlighted as the declared goal of pharmacogenetics. In this paper, the content and significance of the individualization concept are analyzed. METHOD: Our analysis is based on a systematic reading of the current literature pertinent to pharmacogenetics. RESULTS: This analysis reveals that the pharmacogenetic understanding of individualization is based on a biomechanistic paradigm. In contrast to a notion of individualized therapy based on a biopsychosocial paradigm, this biomechanistic concept does not provide for individualization in psychosocial terms, but instead leads to the stratification and classification of patient populations. This finding does not necessarily cast doubt on the efficacy of pharmacogenetics, but does call its underlying ideology into question. CONCLUSION: The term 'individualization of therapy' does not reflect the real potential of pharmacogenetics, but instead represents a widely used and theoretically unjustified publicity slogan. PMID- 14560073 TI - Knowledge and attitudes towards haemophilia: the family side and role of haemophilia societies. AB - OBJECTIVES: A pilot study to evaluate the knowledge about haemophilia in the families enrolled in the Lucknow Haemophilia Society (India), and to assess their attitudes towards prenatal diagnosis (PND). METHODS: A questionnaire to assess the knowledge about haemophilia (questionnaire A) and another questionnaire to assess the attitude towards PND (questionnaire B) were distributed. PND was performed by DNA-based linkage analysis. RESULTS: There was a positive correlation between the knowledge about the disease and the frequency of attending the monthly meetings of the society. The majority of respondents felt that they would opt for termination of pregnancy if the fetus was affected. CONCLUSION: Haemophilia societies act as strong media in educating affected families. Multiple factors affect the attitude towards PND and abortion of affected fetuses. PMID- 14560074 TI - Tackling the problem of rare diseases in public health: the Italian approach. PMID- 14560075 TI - Does early intervention with a light mobilization program reduce long-term sick leave for low back pain: a 3-year follow-up study. AB - STUDY DESIGN: A randomized clinical trial. OBJECTIVES: To evaluate long-term clinical and economical effects of a light mobilization program on the duration of sick leave for patients with subacute low back pain. SUMMARY OF BACKGROUND DATA: Twelve-month follow-up results from a previous study showed that early intervention with examination at a spine clinic, giving the patients information, reassurance, and encouragement to engage in physical activity as normal as possible had significant effect in reducing sick leave. At 12-month follow-up, 68.4% in the intervention group were off sick leave, as compared with 56.4% in the control group. Patients in this study were followed-up for a period of 3 years to investigate possible long-term effects. MATERIALS AND METHODS: Four hundred fifty-seven patients placed on a sick list for 8 to 12 weeks for low back pain were randomized into two groups: an intervention group (n = 237) and a control group (n = 220). The intervention group was examined at a spine clinic and given information and advice to stay active. The control group was not examined at the clinic but was treated within the primary health care. RESULTS: Over the 3 years of observation, the intervention group had significantly fewer days of sickness compensation (average 125.7 d/person) than the control group (169.6 d/person). This difference is mainly caused by a more rapid return to work during the first year. There was no significant difference for the second or third year. In particular, there is no increased risk for reoccurrence of illness from early return to work. At 6-month follow-up, patients in the intervention group were less likely to use bed rest and more likely to use stretching and walking to cope with their back pain compared with the control group. This effect diminished. At 12-month follow-up, the only significant difference between the groups was in the use of stretching. Economic returns of the intervention were calculated in terms of increases in the net present value of production for the society because of the reduction in number of days on sick leave. Net benefits accumulated over 3 years of treating the 237 patients in the intervention group amount to approximately 2,822 dollars per person. CONCLUSIONS: For patients with subacute low back pain, a brief and simple early intervention with examination, information, reassurance, and encouragement to engage in physical activity as normal as possible had economic gains for the society. The effect occurred during the first year after intervention. There were no significant long-term effects of the intervention. The initial gain obtained during the first year does not lead to any increased costs or increased risks for reoccurrence of illness over the next 2 years. PMID- 14560077 TI - Anatomic study of the morphology of human cervical facet joint. AB - STUDY DESIGN: Geometrical properties of the facet joint including cartilage thickness and gap were obtained using human cadaver cervical spinal columns and cryomicrotomy techniques. OBJECTIVES: To determine the existence of level or gender dependency on facet joint morphology in the human cervical spine. BACKGROUND DATA: Although measurements of the human cervical spine have been reported in literature, to the best of knowledge of the authors, geometrical data on the facet joint structures such as the cartilage are not available. These data are important to understand the anatomy of the cervical spine and the role of the cartilage in sharing the external load during physiologic and traumatic situations. Furthermore, the data will assist mathematical modelers to accurately simulate this component of the cervical facet joint in finite element analysis of the spine. MATERIALS AND METHODS: Six unembalmed human cadaver cervical spinal columns were used. A heavy-duty cryomicrotome was used to obtain the geometrical characteristics. The specimens were sectioned in the sagittal plane at 20- to 40 microm intervals. Geometric properties of the facet joint width, cartilage thickness, and cartilage gap (defined as the distance from the ventral-most or dorsal-most region of the facet joint to the location where the cartilage began to appear) were extracted from the anatomic sections that were midsagittal with respect to the facet joints from occiput to T1 levels. Multiple factorial analysis of variance techniques were used to determine the statistical significance of various geometrical parameters obtained from the anatomic sections. RESULTS: The cartilage gap in the upper cervical spine (UCS) (C1-C2, i.e., UCS, 5.4% +/- 0.8) was lower (P < 0.0001) than the gap in the lower cervical spine (LCS) (C3-C7, i.e., LCS, 16.4% +/- 0.8). The gap at the ventral and dorsal regions was lower in the UCS (ventral 3.8% +/- 0.6, dorsal 7.0% +/- 1.4) than in the LCS (ventral 18.5% +/- 0.9, dorsal 14.2% +/- 1.1) with p values of less than 0.0001 and equal to 0.0004, respectively. Further, the gap in the dorsal region for females (14.7% +/- 1.8) was greater (P = 0.0523) than the gap for males (10.8% +/- 1.1). The overall mean facet cartilage thickness was lower (P = 0.0111) in females (0.6 mm +/- 0.1) than males (0.9 mm +/- 0.2) in the UCS. It was also lower (P = 0.0077) in females (0.4 mm +/- 0.02) than males (0.5 mm +/ 0.03) in the LCS. The facet joint width demonstrated differences only between the UCS and LCS (P < 0.0001), with higher magnitudes in the upper (17.4 mm +/- 0.4) than in the lower (11.3 mm +/- 0.3) region. CONCLUSIONS: Facet joint morphology varies with the regions of the cervical spine (upper vs. lower), gender (male vs. female), and location (dorsal vs. ventral). Because of the lack of intervertebral discs in the UCS region, variations in these geometrical characteristics affect the biomechanical behaviors of the human spine secondary to external loads. Furthermore, the lack of adequate cartilage in females may expose the underlying adjacent subchondral bone to direct stresses during normal physiologic and traumatic loads. PMID- 14560078 TI - An extrapedicular approach to the placement of screws in the thoracic spine: an anatomic and radiographic assessment. AB - STUDY DESIGN: A radiographic and anatomic study of an extrapedicular method of screw placement in cadaveric thoracic spines. OBJECTIVE: To evaluate an alternative method of thoracic vertebral body screw fixation using an extrapedicular screw technique. To evaluate the anatomic safe zones and proper starting point for this alternative approach to the placement of screw fixation in the thoracic spine. SUMMARY OF BACKGROUND DATA: Despite the clinical successes reported with thoracic pedicle screw-rod constructs, controversy exists about the safety of this technique in pedicles that are anatomically too small for transpedicular fixation. An alternative method of extrapedicular screw fixation was evaluated in this present study. MATERIALS AND METHODS: Two fresh, unfixed, adult cadavers were obtained randomly; 6.0-mm AO Synthes pedicle screws were placed using an extrapedicular approach bilaterally from T3 to T10. The screws were placed according to one defined method described later. Computerized tomographic (CT) images were obtained. The position of each screw was analyzed. The cadavers were then dissected with the screw pathway exposed and the relation of the screw to surrounding anatomy documented. RESULTS: All screws did not penetrate the spinal canal. All screws were within the pedicle rib unit and did not penetrate the neural foramen or pleura. CONCLUSION: This study, although only introductory, indicates the potential for extrapedicular vertebral body fixation in the thoracic spine. Biomechanical evaluations are presently being conducted to evaluate the use of extrapedicular thoracic screw fixation. PMID- 14560079 TI - Gene transfer of the catabolic inhibitor TIMP-1 increases measured proteoglycans in cells from degenerated human intervertebral discs. AB - STUDY DESIGN: Cells from degenerated intervertebral discs were transduced with an adenoviral vector delivering cDNA of the catabolic inhibitor, TIMP-1, and alterations in the measured proteoglycan were assessed. OBJECTIVES: To assess the potential of TIMP-1 to favorably modify the proteoglycan content of degenerated intervertebral disc cells. SUMMARY OF BACKGROUND DATA: Gene therapy with anabolic factors has resulted in increased proteoglycan synthesis in intervertebral disc cells. Biochemical analysis of degenerated discs has revealed elevated levels of the catabolic enzymes, matrix metalloproteinase, suggesting an intimate role of these factors in the degenerative process. The use of TIMP-1, an endogenous inhibitor of matrix metalloproteinase, via gene therapy may provide an additional method to alter the degenerative processes occurring in the intervertebral disc. MATERIALS AND METHODS: Degenerated intervertebral disc were isolated from eight patients undergoing elective surgical procedures. Cells were cultured in monolayer and transduced with different concentrations of either an adenoviral tissue inhibitor of metalloproteinase-1 (Ad-TIMP-1) or adenoviral-bone morphogenic protein-2 (Ad-BMP-2) construct. Cells were cultured in a three dimensional pellet and proteoglycan synthesis was assessed via 35S-sulfur incorporation. RESULTS: Gene delivery of TIMP-1 and BMP-2 increased measured proteoglycan synthesis at each concentration assessed. IVD cells treated with Ad TIMP-1 demonstrated an optimal response at a multiplicity of infection (MOI) of 100. Cells treated with Ad-BMP-2 demonstrated a progressive increase in proteoglycan synthesis with increasing viral concentrations. CONCLUSIONS: Successful delivery of the anticatabolic gene, TIMP-1, results in increased measured proteoglycan in cultured degenerated disc cells. This finding supports catabolic inhibition as a promising avenue of research for the treatment of degenerative disc disease via gene therapy. PMID- 14560080 TI - A fibronectin fragment stimulates intervertebral disc degeneration in vivo. AB - STUDY DESIGN: A radiographic, histologic, biochemical, and gene expression study was conducted in vivo in a rabbit model to determine the effect of injection of the N-terminal 30 kDa fibronectin fragment (Fn-f) into the intervertebral disc along with various control substances. OBJECTIVE: To determine if the Fn-f is able to induce disc degeneration in vivo. SUMMARY OF BACKGROUND DATA: Animal models of disc degeneration are crucial to defining the molecular events involved in disc degeneration. Although spontaneous and induced models of disc degeneration have been described, none is ideal for molecular studies. A better understanding of disc degeneration at the molecular level is necessary to promote rational design of therapies for degenerative disc disease. MATERIALS AND METHODS: Thirty-one New Zealand white rabbits underwent injection of Fn-f and control substances into the central region of separate lumbar discs using a fine needle. Euthanasia was performed at the 2-, 4-, 8-, 12-, and 16-week time points and the discs were examined radiographically, histologically, biochemically, and with gene expression. RESULTS: Radiographs demonstrated anterior osteophyte formation at Fn-f-injected disc spaces by the 12-week time point. Histology demonstrated a progressive loss of the normal architecture of the nucleus pulposus and anulus fibrosus over the 16-week study period. A progressive loss of proteoglycans was documented using GAG assay but total collagen did not appear to change appreciably. Gene expression studies demonstrated a significant down regulation of both aggrecan and type II collagen mRNA between the 8- and 16-week time points. CONCLUSION: Fn-f appears to induce a progressive degenerative process within the intervertebral disc after injection that resembles degenerative disc disease. This model has several significant advantages for the study of disc degeneration at the molecular level. Further studies are warranted to elucidate the mechanism and molecular events associated with Fn-f-mediated disc degeneration. PMID- 14560082 TI - Biomechanical comparison of cervical spine reconstructive techniques after a multilevel corpectomy of the cervical spine. AB - STUDY DESIGN: An in vitro biomechanical study of several reconstructive techniques after a two-level cervical corpectomy. OBJECTIVES: To evaluate, compare, and quantitate the stability of several reconstructive strategies (anterior, posterior, or anterior/posterior with or without instrumentation) after a multilevel cervical corpectomy. SUMMARY OF BACKGROUND DATA: Several clinical and biomechanical studies have questioned the stability of stand-alone long-segment anterior plate fixation after a multilevel (>or=2) corpectomy. The large cantilever forces generated within the stabilized construct, particularly at the caudal screw-bone interface, have led to plate and screw dislodgement and the need for further surgical intervention. The addition of posterior segmental instrumentation has been shown to improve overall stability and decrease local stresses on the anterior fusion construct (graft and plate). MATERIALS AND METHODS: Seven fresh-frozen cadaveric human cervical spines (C1-T1) were harvested. The C1-C2 and C7-T1 vertebral bodies were embedded in poly methylmethacrylate (PMMA). Three VICON cameras tracked three-dimensional segmental motions at the ends of the fusion construct after a two-level corpectomy and placement of a strut graft with or without instrumentation. Pure moments (flexion/extension, lateral bending, and axial rotation) were applied to the C1 level of each specimen. The motion segments were loaded to a maximum of 2 Nm using dead weights. Testing was first performed on the intact specimens. Then, a two-level corpectomy at the C4 and C5 levels was performed. A PMMA strut graft was then placed into the corpectomy site. Biomechanical testing was then repeated among three different reconstruction techniques: 1) anterior cervical locking plate (PEAK; Depuy-Acromed, Raynham, MA) with dual unicortical screw fixation at C3 and C6; 2) posterior cervical instrumentation (Summit; Depuy-Acromed) using a 3.0-mm rod with segmental lateral mass screw fixation from C3 to C6; and 3) a combined anterior-posterior instrumentation using the anterior PEAK plate and posterior Summit rod system. RESULTS: In all pure moments tested (flexion/extension/lateral bending/axial rotation) the combined anterior posterior instrumentation reconstruction model and the posterior-only instrumentation model were significantly more rigid than the anterior-only instrumentation model (P < 0.05). Interestingly, no statistically significant difference was noted between the combined anterior plate/posterior instrumentation model and the posterior instrumentation-only model. CONCLUSION: The biomechanical results obtained suggest that posterior segmental instrumentation confers significant stability to a multilevel cervical corpectomy regardless of the presence or absence of anterior instrumentation. In cases in which the stability of a multilevel reconstruction procedure is tenuous, the surgeon should strongly consider the placement of segmental posterior instrumentation to significantly improve the overall stability of the fusion construct. PMID- 14560081 TI - Bridging a spinal cord defect using collagen filament. AB - STUDY DESIGN: A rat model of spinal cord defect was designed to evaluate the effect of collagen filament implant on nerve regeneration in the spinal cord defect. OBJECTIVES: To bridge a spinal cord defect and restore the function in adult mammals. SUMMARY OF BACKGROUND DATA: Resection of the spinal cord in mammals is always followed by motor paralysis and loss of voluntary function below the lesion. Partial success in bridging the ends of the spinal cord after complete resection was reported. However, restoration of function has not been reported in adult mammalian. MATERIALS AND METHODS: Four thousand collagen filaments 5-mm-long were grafted to bridge a 5-mm defect of rat spinal cord. Controls had their spinal cord defect left ungrafted after resection. At 1-week intervals, animals were evaluated functionally. After 4 and 12 weeks, animals were evaluated histologically. After 12 weeks, animals were evaluated electrophysiologically. RESULTS: The severed spinal cord axons regenerated along the collagen filament implant crossing the proximal and distal spinal cord implant interfaces at 4 weeks after surgery. The rats with collagen filament grafts could walk, run, and climb with hind forelimb coordination at 12 weeks after surgery. Sensory-evoked potential waveform was found in the rats with collagen filament at 12 weeks after surgery. CONCLUSIONS: The collagen filaments support the axonal regeneration of the transected spinal cord and the restoration of function. PMID- 14560084 TI - Comparison of three lumbar orthoses using motion assessment during task performance. AB - STUDY DESIGN: Four conditions (three orthoses, one no orthosis), full cross-over, randomized order, 12 subjects tested 3 months after a lumbar surgical arthrodesis. OBJECTIVES: To assess whole torso and lumbar motions and comfort for each orthoses condition during performance of activities of daily living. SUMMARY OF BACKGROUND DATA: Previous noninvasive studies measured whole torso (not just lumbar) movement. Recent development of a low profile, flexible, strain-gauged device ("Lordosimeter") facilitated this study. MATERIALS AND METHODS: The Lordosimeter was taped to the skin along the midline of the lumbosacral region. Orthoses studied were Boston anterior opening, Aspen lumbosacral, and Cybertech. Specified activities of daily living were performed for each orthosis condition. Subject effort during the flexion/extension task was assessed by surface electromyography (EMG). Comfort was assessed by visual analog scale. RESULTS: There were no significant differences in EMG between orthoses. For lumbar flexion, trunk flexion, total lumbar motion, and total trunk motion, the mean values for Aspen and Boston (but not Cybertech) were significantly smaller than for no orthosis for most of the tasks and there were no significant differences between Aspen and Boston. The total lumbar motion allowed by each orthosis (averaged across tasks and relative to the no orthosis condition) was 81% for Aspen, 79% for Boston, and 97% for Cybertech. The comfort rating averaged across tasks was 2.24 for Aspen, 4.12 for Boston, and 3.92 for Cybertech (0 = very comfortable, 10 = very uncomfortable). Aspen was significantly more comfortable than Boston or Cybertech. Boston and Cybertech did not differ significantly. CONCLUSIONS: The Lordosimeter is useful for measuring lumbar motion during orthosis wear. The Aspen and the Boston orthoses provided significant flexion extension motion restriction compared with no orthosis, but for almost all of the motion measures did not differ from each other. The Cybertech did not differ significantly from the no orthosis condition. The Aspen orthosis was rated significantly more comfortable than the Boston or the Cybertech. PMID- 14560085 TI - Intraoperative biomechanical assessment of lumbar spinal instability: validation of radiographic parameters indicating anterior column support in lumbar spinal fusion. AB - STUDY DESIGN: Lumbar spinal instability was evaluated using radiographic parameters and intraoperative biomechanical measurement. OBJECTIVES: To validate a conventional radiographic assessment using an intraoperative biomechanical measurement and to determine the radiographic parameters indicating anterior column support in lumbar spinal fusion. BACKGROUND: Posterior lumbar interbody fusion is gaining acceptance in the treatment of degenerative spondylolisthesis, whereas posterolateral fusion alone may result in good clinical outcomes. A quantitative biomechanical assessment of spinal instability is essential to determine the optimal method of spinal arthrodesis. MATERIALS AND METHODS: Nineteen cases with symptomatic degenerative spondylolisthesis were included in this study. All were candidates for pedicle-screw instrumented lumbar arthrodesis. A total of five L3-L4 segments and seventeen L4-L5 segments were investigated. Radiographic assessment included disc angle, range of motion (ROM), percent of slip, and percent of posterior disc height. Distraction stiffness of the operative segment was measured intraoperatively using a strain-gauged spreader. RESULTS: Disc angle in flexion and ROM were highly correlated to distraction stiffness. Severity of slip, disc space narrowing, and facet tropism did not influence the distraction stiffness. The cases with segmental kyphosis in flexion had significantly lower distraction stiffness than those showing segmental lordosis in flexion. CONCLUSIONS: Disc angle in flexion and ROM were the most prognostic parameters of lumbar distraction instability. Although the option of spinal arthrodesis method should be determined based on both clinical manifestation and imaging studies, the current study demonstrated that providing of anterior column support is biomechanically reasonable for degenerative spondylolisthesis with segmental kyphosis in flexion. PMID- 14560086 TI - Three-dimensional analysis of the movement of lumbar spinal nerve roots in nonsimulated and simulated adhesive conditions. AB - STUDY DESIGN: Biomechanical analysis of the movement of the lumbar spinal nerve roots (NRs). OBJECTIVES: The characteristics of the movement of lumbar spinal NRs corresponding to the change of posture were three-dimensionally analyzed in nonsimulated and simulated adhesion conditions using a porcine model. SUMMARY OF BACKGROUND DATA: There is a paucity of data on the movement of NRs. MATERIALS AND METHODS: Thirty two NRs of four pig cadavers were investigated. (study 1) Two tantalum beads were sutured on respective NRs from L3 to L6. Five beads were put into each bony structure as reference points. The porcine body was put on the cage designed for three-dimensional radiostereometric analysis. Two oblique radiographs were simultaneously taken at flexion, neutral, and extension. (study 2) After study 1, either L4 NR was ligated onto the adjacent bone to mimic monoradicular adhesion. Radiographs were taken in the same manner. The movements of the beads on NRs relative to immobile standard points in each vertebra were analyzed. RESULTS: The movement of NRs was characterized as a combination of stretching/slackening and pendulum motion. The distance that NRs were stretched/slackened, the excursion, and the angles that NRs moved were: 0.39 +/- 0.29 mm, 1.81 +/- 0.63 mm, and 10.46 +/- 5.93 degrees (in nonadhesion), and 0.30 +/- 0.20 mm, 1.02 +/- 0.50 mm, and 7.13 +/- 3.79 degrees (in simulated adhesion), respectively. CONCLUSION: Adhesion might play a part in the pathogenesis of lumbar spinal disorders because mechanical irritation secondary to stretching/slackening and pendulum motion might be concentrated at the distal part of NRs, adjacent to adhesion site, which has less cerebrospinal fluid and is close to dorsal root ganglion. PMID- 14560087 TI - Predictive value of pelvic incidence in progression of spondylolisthesis. AB - STUDY DESIGN: Retrospective analysis of pelvic incidence and other radiographic parameters as a predictor of progression of isthmic spondylolisthesis. OBJECTIVES: To evaluate the predictive value of various radiographic parameters, including pelvic incidence, in determining the risk for progression of lumbosacral isthmic spondylolisthesis. SUMMARY OF BACKGROUND DATA Although pelvic incidence has recently been shown to be positively correlated to the severity of spondylolisthesis, it has not been confirmed as a predictor of spondylolisthetic progression. MATERIALS AND METHODS: Thirty-six patients who have undergone primary posterior lumbosacral fusion for isthmic spondylolisthesis at our institution from 1977 to 2001 were retrospectively analyzed. There were 24 females and 12 males with a mean age of 21.3 +/- 2.0 years (range, 12 to 53 y). Twenty-two patients had high-grade (Meyerding class III, IV, V) and 14 patients had low-grade (Meyerding class I, II) spondylolisthesis, respectively. Factors evaluated included age, gender, neurologic deficits, reason for surgery, and documented evidence of progression. Slip percentage, high-grade or low-grade slip, slip angle, sacral inclination, sacral rounding, trapezoidal L5 vertebra, and pelvic incidence were measured from immediate preoperative standing lateral radiographs. These factors were statistically analyzed for risk of progression. Continuous variables were analyzed using one-way analysis of variance. Nominal variables were analyzed using chi2 test. RESULTS: Pelvic incidence (P = 0.66) was not predictive of spondylolisthetic progression. Of the other radiographic measurements, slip percentage (P < 0.001), slip angle (P = 0.016), and high-grade spondylolisthesis (P < 0.0001) were highly predictive of progression. Interestingly, sacral inclination (P = 0.33) was not predictive of progression. CONCLUSIONS: Pelvic incidence cannot adequately predict the probability of spondylolisthetic progression. Analysis of the other clinical and radiographic parameters revealed that slip percentage and high-grade spondylolisthesis remain the most positive predictors of progression. PMID- 14560089 TI - Is local bone viable as a source of bone graft in posterior lumbar interbody fusion? AB - STUDY DESIGN: Radiographic evaluation of bony union and clinical outcome were investigated after posterior lumbar interbody fusion (PLIF) performed with only local bone. OBJECTIVES: To examine the viability of local bone for bone graft in PLIF. SUMMARY OF BACKGROUND DATA: Different sources of bone graft have been used for PLIF. Autologous iliac bone is most likely suitable for biologic bone healing ability, but there may be some questions such as donor site pain. Allograft and artificial bones are less effective for bone healing. Use of local bone could be a solution. However, the quality of local bone is still being questioned and there is hesitation to use local bone alone. MATERIALS AND METHODS: Thirty-two patients underwent PLIF (24 single-level and 8 double-level) using the Brantigan I/F Carbon Cage filled with local bone and segmental pedicle screw fixation. At 3, 6, and 12 months after surgery, "parallel AP view radiograph" was obtained to evaluate bony union status. Average follow-up period was 26 months. Clinical outcome was also evaluated by the Japanese Orthopedic Association clinical score. RESULTS: The bony union rate was 16.7%, 72.4%, and 100% at 3, 6, and 12 months after surgery, respectively. Clinically, the average of JOA scores was improved significantly (P < 0.001). The improvement rate was 72.9% (12 mo) and 76.8% (24 mo) on average. CONCLUSIONS: A 100% bony union rate was obtained 12 months after PLIF with only local bone. The results showed that clinical viability of local bone as a source of bone graft in PLIF. PMID- 14560090 TI - Orthopaedic implications in the management of pediatric vertebral and spinal cord tumors: a retrospective review. AB - STUDY DESIGN: Retrospective review of pediatric patients with vertebral and spinal cord tumors. OBJECTIVES: To identify clinical and radiographic features of diagnostic importance, and to evaluate the effectiveness of surgical treatment with respect to complications in pediatric patients with vertebral and spinal cord tumors. SUMMARY OF BACKGROUND: Vertebral and spinal cord tumors in children are uncommon and present a significant challenge in diagnosis, treatment, and management of complications. Because most signs and symptoms of these tumors are musculoskeletal in nature, it is imperative for orthopedic surgeons to be familiar with them. MATERIALS AND METHODS: Medical charts and radiographic records of 26 pediatric patients with documented vertebral and spinal cord tumors were reviewed retrospectively. RESULTS: There were 10 malignant and 16 benign tumors. Mean age at diagnosis was 10 years, and the male-to-female ratio was 1:1. Mean clinical follow-up was 25.8 months. The average delay in diagnosis was 10.9 months. The two most common reasons for presentation were pain (n = 20, 76%) and neurologic involvement (n = 14, 54%). Initial plain radiographs were positive in 72% of patients, whereas MRI showed the tumor in 96% of studies. Sixteen patients underwent spinal fusion after excision of the tumor. There were no perioperative deaths. Complications were common, with an average of 1.1 complications per patient. At latest follow-up, six patients had neurologic deficits, two patients had died, and one patient was terminally ill. CONCLUSION: Increased awareness of modes of presentation, use of modern imaging techniques, and an aggressive approach to spinal stabilization at the time of laminectomy should decrease morbidity in patients with vertebral and spinal cord tumors. PMID- 14560091 TI - Respiratory function and cosmesis at maturity in infantile-onset scoliosis. AB - STUDY DESIGN: Retrospective review of patient records, clinical and radiographic, and patient recall for full pulmonary function studies and surface topography. OBJECTIVES: Assessment of outcome of treatment policy after age 15 during the previous 30 years to establish the efficacy of management protocols in a group that is too small and too varied for more formal assessment. BACKGROUND DATA: Spinal deformity presenting during infancy or early childhood poses a clinical problem caused by small numbers, long growth period, variable presentation and treatment methods, and, finally, the length of time that must pass before meaningful outcome results can be assessed. The aims of treatment are to preserve respiratory function and cosmetic appearance. MATERIALS AND METHODS: The records of patients with infantile onset, nonsyndromic, and noncongenital scoliosis were reviewed. Thirty two were at least age 15 years at the time of review and 21 of these agreed to attend for full pulmonary function testing (spirometry, lung volumes, gas diffusion) and surface topography, whereas two more had recent spirometry results available in their record. Treatment had been serial casting with Risser jacket, bracing, or surgery. RESULTS: Those whose scoliosis resolved or was stabilized by nonoperative means (N = 6) at an acceptable Cobb angle had normal cosmesis and pulmonary function (mean FEV1 = 98.7%, mean FVC = 96.6%). Those who were managed by casting or bracing and underwent surgery after age 10 (N = 6, mean age at surgery 12.9 y) had variable cosmesis and acceptable pulmonary function (mean FEV1 = 79%, mean FVC = 68.3%). Those whose deformity necessitated early surgery (N = 11, mean age at surgery 4.1 y) had recurrence of deformity and diminished respiratory function (mean FEV1= 41%, range 14%-72%, mean FVC = 40.8%, range 12%-67%). CONCLUSIONS: Although these are small numbers and treatment methods have changed since the beginning of the series, the results indicate that this condition is not simple to treat and for some children still has the risk for serious deformity and respiratory compromise. There is, as yet, no evidence that early surgical intervention in this group of patients with infantile scoliosis has altered their prognosis in any meaningful way. PMID- 14560092 TI - Physical functioning: self-report and performance measures are related but distinct. AB - STUDY DESIGN: Cross-sectional study of 63 patients with chronic low back pain (CLBP). OBJECTIVES: To determine the relationship between the SF-36 physical functioning (PF) scale and physical performance on a modified symptom limited treadmill test, and to determine the effect of disability status and SF-36 mental health (MH) scores on self-reported physical functioning and on physical performance. SUMMARY OF BACKGROUND DATA: Patients with CLBP report low levels of physical functioning. Because self-report measures are influenced by psychosocial factors, such as disability and low MH, a common recommendation is to supplement self-report with objective or physical performance testing. Little information exists regarding whether performance testing is also influenced by disability and low MH. MATERIALS AND METHODS: Patients completed the SF-36 and performed a maximal, symptom-limited, modified treadmill test. Total walking time and heart rate on the treadmill were registered. The effects of disability and low MH on self-report and performance testing were calculated by unpaired Student's t testing and effect size estimation. RESULTS: A strong relationship between the PF scale of the SF-36 and treadmill walking time was found, but there was little evidence of overlap between these physical functioning domains. Both disability status and low MH had a much larger effect on self-report measures than on physical performance. CONCLUSIONS: Self-report measurements and performance-based assessments provide information about distinct, although related, domains of physical functioning. Disability and low MH are associated with lower SF-36 PF scores. Our results confirm that self-report measures require supplementation with objective performance testing to provide optimal assessment for patients with CLBP. PMID- 14560093 TI - Validation of the Japanese version of the Roland-Morris Disability Questionnaire for Japanese patients with lumbar spinal diseases. AB - STUDY DESIGN: This study was designed to validate a translated version of the Roland-Morris Disability Questionnaire (RMDQ). OBJECTIVE: To validate the Japanese version of the RMDQ (JRMDQ) for Japanese patients with low back pain (LBP) and/or lumbar spinal diseases (LD). SUMMARY OF BACKGROUND DATA: To assess the effect of interventions, a scale that directly assesses disabilities in patients with LBP and/or LD and that can be used internationally needs to be established in Japan. MATERIALS AND METHODS: To promote the JRMDQ, the RMDQ was translated into Japanese and then back-translated. In four hospitals, 320 patients with LBP and/or LD were examined. At first, they were examined using the JRMDQ, the Japanese Orthopedic Association (JOA) score, and the visual analogue scale (VAS). The repeatability of the JRMDQ was evaluated in 55 patients by two examinations within 2 weeks. To examine the validity of the JRMDQ, the correlation between the JRMDQ and other scales was calculated in all 320 patients. RESULTS: The JRMDQ demonstrated significant repeatability (R = 0.810, P < 0.0001). The JRMDQ and other scales demonstrated significant correlations (JRMDQ-JOA, R = -0.772; JRMDQ-ADL items of JOA score, R = -0.790; JRMDQ-VAS, R = 0.447, P < 0.0001). CONCLUSIONS: The JRMDQ is a repeatable and valid questionnaire for assessing disabilities caused by LBP and/or LD. The JRMDQ is equivalent to the English RMDQ. PMID- 14560094 TI - Sensory stimulation-guided sacroiliac joint radiofrequency neurotomy: technique based on neuroanatomy of the dorsal sacral plexus. AB - STUDY DESIGN: A retrospective audit and examination of anatomic findings. OBJECTIVE: To examine the effectiveness of sensory stimulation-guided radiofrequency neurotomy for the treatment of recalcitrant sacroiliac joint pain. SUMMARY OF BACKGROUND DATA: Sacroiliac joint-mediated pain is a distinct clinical entity. The prevalence of intra-articular pain arising from the sacroiliac joint in patients with low back pain has been estimated at 15% to 30%. Unfortunately, the clinical success of current treatment methods for chronic sacroiliac pain is discouraging. Based on the anatomy of the sacral posterior primary rami and their lateral branch nerves, an anatomically based sensory stimulation-guided radiofrequency technique was developed to overcome the inherent challenge posed by the variable topography of the sacral lateral branch nerves. MATERIALS AND METHODS ANATOMIC STUDY: Meticulous dissection exposing the dorsal sacral plexus and lateral branch nerves entering the sacroiliac joint complex was performed on three cadaveric specimens. Small-gauge wires were placed adjacent to the lateral branch nerves entering the joint and over the dorsal sacrum to the dorsal sacral foramina. Fluoroscopic images were obtained correlating the location and number of these branches arising from the posterior primary rami of S1-S3 to identifiable bony landmarks. CLINICAL STUDY: A retrospective chart review was performed selecting patients who underwent sensory stimulation-guided sacral lateral branch radiofrequency neurotomy after dual analgesic sacroiliac joint deep interosseous ligament analgesic testing between February 17, 1998 and March 15, 1999. RESULTS: A total of 14 patients met inclusion criteria for this retrospective study. Success was defined as greater than 60% consistent subjective relief and greater than a 50% consistent decrease in visual integer pain score, maintained for at least 6 months after the procedure. Sixty-four percent of patients experienced a successful outcome, with 36% experiencing complete relief. Fourteen percent of patients did not achieve any improvement. No patients experienced a complication or worsening of their pain from the procedure. CONCLUSIONS: A sensory stimulation-guided approach toward the identification and subsequent radiofrequency thermocoagulation of symptomatic sacral lateral branch nerves appears to offer significant therapeutic advantages over existing therapies for the treatment of chronic sacroiliac joint complex pain. PMID- 14560095 TI - Carl Nicoladoni and his contributions to scoliosis. AB - Carl Nicoladoni (1847-1902) studied medicine in Vienna and became Privatdozent in surgery in 1876. He accepted a chair as a Professor of Surgery at the university clinics of Innsbruck (1881) and Graz (1895). Nicoladoni has made significant contributions in the progress of surgery and performed a variety of operations in several surgical disciplines. However, his principal contributions are in the field of orthopedic surgery, in particular his excellent and detailed studies on scoliosis, based on thorough anatomic and kinetic investigation. His commitment to the research of scoliosis resulted in three books and three bigger treatises, all of which were printed in German. His earliest book was printed in 1882 and was on the torsion of the scoliotic spine, Die Torsion der Skoliotischen Wirbelsaule. Three bigger treatises on scoliosis entitled Die Architektur der Sskoliotischen Wirbelsaule (The Architecture of the Scoliotic Spine, 1889), Die Architektur der Kindlichen Skoliose (The Architecture of Juvenile Scoliosis, 1894), and Die Skoliose des Lendensegmentes (The Scoliosis of Lumbar Segments, 1894) were published in an anthology called Denkschriften der Kaiserlichen Akademie der Wissenschaften, a series of publications from the imperial academy of science. Two versions of his epical work, Anatomie und Mechanismus der Skoliose (Anatomy and Mechanism of Scoliosis) were printed. A larger edition printed in 1904 was part of the Bibliotheca Medica, a monumental series of various clinical books published around the beginning of the past century. The second version, a shortened one, with the same title was included in an anthology called Deutsche Chirurgie (German Surgery) and published in 1909. The purpose of this historical article is to discuss Nicoladoni's achievements in the field of scoliosis based on a detailed analysis of his books. PMID- 14560096 TI - Altered subarachnoid space compliance and fluid flow in an animal model of posttraumatic syringomyelia. AB - STUDY DESIGN: A histologic study of cerebrospinal fluid tracers in Sprague-Dawley rats undergoing lumboperitoneal shunt insertion in the excitotoxic animal model of posttraumatic syringomyelia (PTS). OBJECTIVES: To determine the effects of cerebrospinal fluid (CSF) diversion from the subarachnoid space on perivascular flow (PVS) and syrinx formation in posttraumatic syringomyelia. SUMMARY OF BACKGROUND DATA: In an animal model of PTS, fluid enters syringes from the subarachnoid space via perivascular spaces. Preferential PVS flow occurs at the level of the syrinx. It has been suggested that arachnoiditis predisposes to posttraumatic syringomyelia formation by obstructing subarachnoid cerebrospinal fluid flow and enhancing perivascular flow. MATERIALS AND METHODS: Thirty-two male Sprague-Dawley rats were investigated using the CSF tracer horseradish peroxidase (HRP), the excitotoxic model of PTS, and lumboperitoneal shunt insertion. Five experimental groups consisted of normal controls, syrinx only and shunt only controls, and shunt insertion before or after syrinx formation. In all groups except normal controls, CSF flow studies were performed 6 weeks after the final intervention. Grading scales were used to quantify HRP staining. RESULTS: All excitotoxic model animals formed syringes. Perivascular flow was greatest at the level of the syrinx. Cerebral cortex perivascular flow was significantly reduced after shunt insertion in animals with a syrinx (P < 0.05). Shunt insertion did not alter syrinx length or size. There were no significant differences between shunt and syrinx first groups. CONCLUSIONS: Increasing caudal subarachnoid space compliance with a shunt does not affect local CSF flow into the spinal cord and syrinx. These results suggest that localized alterations in compliance, as opposed to obstruction from traumatic arachnoiditis, may act as an important factor in syrinx pathogenesis. PMID- 14560097 TI - Intradural extramedullary mass formation in spinal cord sarcoidosis: case report and literature review. AB - STUDY DESIGN: A case of myelopathy caused by intradural extramedullary spinal cord sarcoidosis. OBJECTIVES: To report a rare case of mass formation in spinal cord sarcoidosis and discuss the diagnostic approach and therapeutic management of this condition. SUMMARY OF BACKGROUND DATA: Spinal cord sarcoidosis is a rare condition that is difficult to diagnose, and intradural extramedullary mass formation is even rarer. MATERIALS AND METHODS: A 42-year-old woman first noticed numbness in her fingers, and these symptoms gradually spread to her hands and feet. MRI revealed an intradural extramedullary mass, as confirmed by an isointensity T1 and low-intensity T2 signal, and also enhanced by Gd-DTPA. The authors suspected the mass to be a meningioma and performed surgery to resect it. RESULTS: The mass was observed through the dura, and it was twined with the rootlet. After surgery, the patient experienced temporary paralysis of her right upper extremity, followed rapidly by almost complete neurologic recovery. Bilateral hilar lymphadenopathy was noted and this was diagnosed histologically as lung sarcoidosis. One year after surgery, a complete AV block occurred because of heart sarcoidosis and a pacemaker was inserted. CONCLUSIONS: Spinal cord sarcoidosis is rare disease and is difficult to diagnose, but when a mass is present in the spinal canal, it is important to remain open to the possibility of spinal cord sarcoidosis. Besides if it is diagnosed as spinal cord sarcoidosis, it is also important to remain systemic sarcoidosis before, during, and after surgery to check general conditions and to guide appropriate treatment. PMID- 14560098 TI - Successful management of a large pulmonary cement embolus after percutaneous vertebroplasty: a case report. AB - Percutaneous vertebroplasty is increasingly used for the treatment of vertebral compression fractures. Local leakage of polymethylmethacrylate cement into the perivertebral space is a common complication, but important systemic effects have rarely been reported. The authors describe the case of a 52-year-old patient with central pulmonary embolism after percutaneous vertebroplasty of the eleventh thoracic vertebral body. The large cement embolus was removed from the right pulmonary artery with a hybrid technique combining an interventional catheter procedure with an open heart operation. The patient made an uneventful recovery. The authors review how appropriate arthroplasty techniques might minimize the risk of this dreadful complication. PMID- 14560099 TI - Presacral multiple cellular schwannomas. AB - STUDY DESIGN: Case report. The authors documented a patient presenting with multiple cellular schwannomas along the sciatic nerve. The patient had no stigmas of neurofibromatosis. Thirteen tumors were resected via both anterior and posterior approaches. OBJECTIVES: To point out that presacral cellular schwannoma can display multiple occurrences. SUMMARY OF BACKGROUND DATA: Cellular schwannoma, which is a well-recognized variant of benign schwannoma, is often misdiagnosed as sarcoma. The tumor is usually a solitary lesion. MATERIALS AND METHODS: Medical history, physical findings, imaging features, and histologic findings were reviewed in a case of multiple cellular schwannomas in the presacral regions. RESULTS: A 39-year-old woman without any evidence of neurofibromatosis underwent removal of presacral multiple tumors along the sacral nerve root. Nineteen years after the surgery, the tumors recurred with multiple fashion. Thirteen tumors were resected via both anterior and posterior approaches. Immunohistochemical analysis of S-100 protein and Ki-67 were beneficial with respect to differentiation of cellular schwannoma from malignant peripheral nerve sheath tumor. CONCLUSIONS: The present case illustrated that multiple cellular schwannomas can develop in nonneurofibromatosis patients. PMID- 14560100 TI - Intradural venous varix: a rare cause of an intradural lumbar spine lesion. AB - STUDY DESIGN: A case of intradural venous varix producing a mass-like lesion in the lumbar spine is reported. OBJECTIVE: To present a rare cause of an intradural mass lesion along the cauda equina. To present radiologic findings of this entity and to offer a hypothesis for its formation. SUMMARY OF BACKGROUND DATA: Epidural venous varices have been described previously in the literature. To the authors' knowledge, no cases of intradural venous varix presenting as an intradural mass have been reported previously. MATERIALS AND METHODS: An elongated, serpentine intradural mass was discovered on routine lumbar MRI performed for investigation of the patient's radiculopathy. The patient underwent laminectomy and intradural exploration. RESULT: On surgical exploration, a serpentine, massively dilated vein was identified, which distorted the normal nerve roots. CONCLUSION: A venous varix should be considered in the differential diagnosis of an intradural lesion in the lumbar spine in the appropriate clinical setting. PMID- 14560102 TI - Sepsis and the dendritic cell. AB - Sepsis is a syndrome of significant morbidity and mortality. Unlike the advances made in other diseases processes, improvements in outcome from sepsis, severe sepsis, and septic shock have been modest. Current research has altered our understanding of sepsis pathogenesis such that present models and definitions are still evolving. One relatively novel cell type, the dendritic cell, is the subject of much current investigation in sepsis. Although our present understanding of dendritic cell biology is incomplete, growing evidence supports the importance of this antigen-presenting cell in the normal and maladaptive responses to microbial invasion and tissue injury. A better understanding of this cell's basic biology as well as its potential as a therapeutic target will undoubtedly play increasing roles in the development of new strategies for the treatment of the septic patient. PMID- 14560103 TI - Peptidoglycan and lipoteichoic acid in gram-positive bacterial sepsis: receptors, signal transduction, biological effects, and synergism. AB - In sepsis and multiple organ dysfunction syndrome (MODS) caused by gram-negative bacteria, lipopolysaccharide (LPS) initiates the early signaling events leading to the deleterious inflammatory response. However, it has become clear that LPS can not reproduce all of the clinical features of sepsis, which emphasize the roles of other contributing factors. Gram-positive bacteria, which lack LPS, are today responsible for a substantial part of the incidents of sepsis with MODS. The major wall components of gram-positive bacteria, peptidoglycan and lipoteichoic acid, are thought to contribute to the development of sepsis and MODS. In this review, the literature underlying our current understanding of how peptidoglycan and lipoteichoic acid activate inflammatory responses will be presented, with a focus on recent advances in this field. PMID- 14560104 TI - Modulation of the lipopolysaccharide receptor complex (CD14, TLR4, MD-2) and toll like receptor 2 in systemic inflammatory response syndrome-positive patients with and without infection: relationship to tolerance. AB - The lipopolysaccharide (LPS) receptor complex consists of two interacting receptors (CD14 and TLR4) and an associated protein (MD-2). When engaged by LPS, as in gram-negative infection, this complex transduces a signal detected by MyD88 and passed onward by a cascade of the IRAKs, TRAF6, and NIK, resulting in activation of NF-kappaB. A similar cascade, mediated by TLR2, occurs with ligands derived from gram-positive bacteria. In vitro studies of human monocytes have shown that TLR4 mRNA is paradoxically upregulated in response to "tolerizing" doses of LPS. This study evaluated changes in vivo of blood monocyte CD14, TLR4, TLR2, and MD-2 mRNA by reverse transcription followed by real-time polymerase chain reaction in surgical intensive care unit patients and in normal controls. In addition cell-surface receptor expression of TLR2, TLR4, and CD14 was assessed by flow cytometry in patients and normal controls. Inflammation-induced acute tolerance to LPS was evaluated by ex vivo whole blood tumor necrosis factor alpha production and was significantly reduced in patients compared with controls, confirming LPS hyporesponsiveness. Monocyte mRNA and cell-surface receptor expression of TLR4 were increased 2.4-fold (P < 0.05) and 1.7-fold (P <.002), respectively, in patients compared with normal controls. Monocyte TLR2 mRNA, MD-2 mRNA and CD14 and TLR2 cell-surface expression were not significantly changed compared with controls. The present study suggests that the acute inflammatory condition associated with peripheral cellular LPS hyporesponsiveness is neither specific to prior infectious challenge nor can be ascribed to significant alterations in expression of the cell-surface LPS binding complex proteins. PMID- 14560105 TI - The clinical value of procalcitonin and neopterin in predicting sepsis and organ failure after major trauma. AB - We examined whether procalcitonin (PCT) or neopterin (NT) are useful in predicting sepsis, multiple organ failure (MOF), or death after multiple trauma (MT). In a prospective clinical study, a total of 137 consecutive trauma patients (mean age 39 years, median injury severity score [ISS] 27 points) and 34 healthy volunteers were enrolled. Blood samples were collected on arrival in the emergency room until day 28 after trauma. Plasma NT was detected by enzyme-linked immunoassay and PCT plasma levels were determined using an immunoluminometric assay. The incidence of sepsis was 65%, MOF 48%, and death in hospital within 28 days 11%. After adjustment for age, gender, and ISS, PCT and NT levels during the first 2 days after injury were unable to differentiate between patients who developed sepsis or not. On the contrary, patients who developed MOF had higher PCT plasma levels on day 0 (0.60 vs. 0.15 ng/mL), and on days 1 and 2 combined (1.95 vs. 0.32 ng/mL). This difference remained significant in multivariate logistic regression (P = 0.01) and additional subgroup analyses for early and late MOF (P = 0.048 and 0.002). For NT, smaller differences were observed (4.39 vs. 3.68 nmol/L, and 7.20 vs. 5.79 nmol/L), which lost significance in multivariate analysis. On the basis of PCT, ISS, and age, a MOF prediction rule was developed and had a good predictive power (area under the curve: 0.77; P < 0.001). These findings demonstrate that high plasma concentrations of PCT in the early posttraumatic phase are an independent predictor of MOF but not of sepsis. PMID- 14560106 TI - First use of hypertonic saline dextran in children: a study in safety and effectiveness for atrial septal defect surgery. AB - Hypertonic saline dextran (7.5% NaCl + 6% Dextran-70) has been used in adults in several studies and shown beneficial effects in hypovolemic shock, trauma, cardiogenic shock, and cardiac surgery. There have never been studies of this solution in children. This work studies its effect in children undergoing surgery for the correction of atrial septal defects. Twenty-five children underwent correction of atrial septal defect using cardiopulmonary bypass with bloodless priming. Children were divided in five groups and each received an incremental hypertonic saline dextran dose of 0.1, 0.5, 1.0, 2.0, and 4.0 mL/kg, 5 min before the beginning of cardiopulmonary bypass. Collected data were fluid balance, amount of bleeding, blood/derivative transfusion occurrence, plasma sodium, and hematocrit. Patients were divided into low-dose (0-1 mL/kg) and high-dose (2-4 mL/kg) groups. Analysis of variance was used to determine differences in blood loss between groups. The fluid balance and blood/derivative requirements were compared through Student's t test and Fisher's exact test (2-tail), respectively. All patients were discharged from hospital with corrected atrial septal defect. No hypertonic saline dextran-related complications occurred. There were no differences in the amount of bleeding. The high-dose group exhibited a significant decrease in fluid balance and in blood/derivative requirements in comparison with the low-dose group. In this study, the use of hypertonic saline dextran in the pediatric population submitted to cardiopulmonary bypass is safe and does not raise the amount of bleeding. Its effective doses produce negative fluid balance and reduce blood/derivative requirements. PMID- 14560107 TI - Lipopolysaccharide dose response in baboons. AB - A lipopolysaccharide (LPS) dose-response study in an experimental baboon endotoxemia model is presented to define the relevance of this model compared with human endotoxemia. We describe acute and subacute endotoxemic models in baboons, the first evoked by bolus injection of LPS (1 mg, 0.1 mg, or 4 ng per kg of Escherichia coli LPS), and the second evoked by infusion of 1.5 mg/kg of E. coli LPS over 30 min. We report the analysis of LPS clearance, the kinetics of tumor necrosis factor, interleukin (IL) 6, and IL-8 expression on the protein as well as on the mRNA level, change in blood counts (white and red blood cells and circulating platelets), and several hemodynamic parameters such as temperature, cardiac index, heart rate, and mean arterial pressure via multiple sampling. The resulting data are compared with previously published human data. Our results show that the LPS-induced kinetics of cytokine release, as well as of hemodynamic and hematologic changes in baboons, were similar to those observed in humans, even though baboons required a approximately 104-fold higher initial LPS dose to develop these manifestations. Hence, we demonstrate that endotoxemia in baboons qualitatively, yet not quantitatively, resembles endotoxemia in humans and, therefore, proves to constitute a useful model for studying the pathogenic mechanisms of sepsis in relation to humans. PMID- 14560108 TI - Bcl-2 inhibits gut epithelial apoptosis induced by acute lung injury in mice but has no effect on survival. AB - Gut epithelial apoptosis is increased in human studies and animal models of noninfectious inflammation and sepsis. Elevated intestinal cell death appears to be physiologically significant in sepsis. Previous studies demonstrate that overexpression of the antiapoptotic protein Bcl-2 in the gut epithelium of transgenic mice is associated with improved survival from Pseudomonas aeruginosa pneumonia and cecal ligation and puncture. The functional significance of elevated gut apoptosis in noninfectious inflammation has not been examined. We hypothesized that intestinal apoptosis would be detrimental to survival in noninfectious critical illness. To address this issue, acute lung injury (ALI) was induced with intratracheal injection of lipopolysaccharide (LPS, 800 microg) in wild-type (WT) FVB/N mice and transgenic mice that overexpress Bcl-2 in their intestinal epithelium. Guts were harvested at 12, 24, 48, and 72 h and assessed for apoptosis by both hematoxylin and eosin and active caspase-3 staining in 100 contiguous crypts. ALI increased gut epithelial apoptosis 12 h after LPS instillation compared with shams (P < 0.01), whereas overexpression of Bcl-2 decreased intestinal apoptosis compared with WT animals with ALI when assayed by active caspase-3 (P < 0.05). Plasma levels of tumor necrosis factor alpha, interleukin (IL)-6, and IL-10 were similar between WT and transgenic animals with ALI, both of which had elevated IL-10 levels at 12 h and elevated IL-6 levels at 24 h compared with sham animals. In a separate experiment, transgenic and WT animals with ALI were followed for mortality to determine whether gut overexpression of Bcl-2 conferred a survival advantage. Survival at 10 days was 73% in WT animals (n = 33) and 65% in Bcl-2 animals (n = 23, P = ns). These results indicate that while gut epithelial apoptosis is elevated in multiple models of critical illness, prevention of intestinal cell death by overexpression of Bcl-2 is associated with a disparate survival effect between sepsis and noninfectious inflammation. PMID- 14560109 TI - Improved survival in uncontrolled hemorrhagic shock induced by massive splenic injury in the proestrus phase of the reproductive cycle in the female rat. AB - Estrogen has a protective effect in adverse circulatory conditions such as trauma hemorrhage and ischemia-reperfusion. In the present investigation uncontrolled hemorrhagic shock induced by massive splenic injury (MSI) was studied in male and female rats during the proestrus and metestrus phase of the reproductive cycle. The animals were anesthetized and randomly divided into four groups: group 1 (n = 8) were sham operated female animals; in group 2 (n = 8) MSI was induced in male animals; in group 3 (n = 8) MSI was induced in female animals in proestrus; in group 4 (n = 8) MSI was induced in female animals in metestrus. Plasma 17beta estradiol was 112.8 +/- 16.7 pg/mL in group 3 and 44.8 +/- 16.7 pg/mL in group 4 (P < 0.04). After MSI the mean arterial pressure dropped in group 2 from 135.6 +/ 3.9 to 47.6 +/- 8.8 mmHg (P < 0.001) in 60 min. Total blood loss in 4 h was 29.6 +/- 3.5% of blood volume and mean survival time 161.3 +/- 30.6 min. In group 3 total blood loss in 4 h was 24.4 +/- 5.7% of blood volume and mean survival time 240 +/- 0 min (P < 0.03 compared with group 2). In group 4, total blood loss in 4 h was 29.6 +/- 3.9% of blood volume and mean survival time was 112.3 +/- 28.7 min (P < 0.001 compared with group 3). These results indicate that female rats in the proestrus phase of the reproductive cycle better maintain hemodynamic stability and survival in uncontrolled hemorrhagic shock than female animals in metestrus or males. This difference is attributed to factors other than blood loss. PMID- 14560110 TI - Perflubron emulsion improves hepatic microvascular integrity and mitochondrial redox state after hemorrhagic shock. AB - Hemorrhagic shock is associated with decreased systemic oxygen delivery, but also with impaired microvascular perfusion, which can result in diminished local oxygen availability even in the presence of adequate cardiac output after resuscitation. Beside surgical interventions to control blood loss, transfusion of stored packed red blood cells represents the current standard of care in the management of severe hemorrhagic shock. Because stored red blood cells are less deformable and show a higher O2 affinity that affects the O2 off-load to tissues, perfluorocarbon-based artificial oxygen carriers might improve local O2 delivery under these conditions. To test this, rats were subjected to hemorrhagic shock (1 h, mean arterial pressure [MAP] 30-35 mmHg) and were resuscitated with fresh whole blood, pentastarch, stored red blood cells, perflubron emulsion (2.7 and 5.4 g/kg body weight) together with pentastarch, or stored red blood cells together with 2.7 g/kg perflubron emulsion. Hepatic microcirculation, tissue oxygenation, and mitochondrial redox state were investigated by intravital microscopy. In addition, hepatocellular function and liver enzyme release were determined. After hemorrhagic shock and resuscitation with perflubron emulsion, volumetric sinusoidal blood flow was significantly increased compared with resuscitation with stored red blood cells. Furthermore, resuscitation with perflubron emulsion resulted in higher hepatic tissue PO2 and normalized mitochondrial redox potential, which was accompanied by lessened hepatocellular injury as well as improved liver function. These results indicate that, in this model of hemorrhagic shock, asanguineous fluid resuscitation with addition of perflubron emulsion is superior to stored blood or pentastarch alone with respect to increased local O2 availability on the cellular level. This effect is primarily due to improved restoration of hepatic microcirculatory integrity. PMID- 14560112 TI - Differential effects of a selective inhibitor of soluble guanylyl cyclase on global and regional hemodynamics during canine endotoxic shock. AB - Activation of soluble guanylyl cyclase (sGC) might occur early during septic shock and play a role in the regulation of vascular tone and the redistribution of blood flow. The aim of this study was to assess the effects of sGC inhibition with oxadiazoloquinoxalinone (ODQ) on global and regional hemodynamic parameters in a clinically relevant model of septic shock. Fifteen anesthetized adult mongrel dogs were equipped with femoral and pulmonary artery catheters and ultrasonic flow probes around the mesenteric, femoral and renal arteries. The animals were randomized to receive Escherichia coli endotoxin (2 mg/kg, i.v.) alone, endotoxin followed by ODQ (1 mg/kg i.v.), or ODQ alone. Endotoxin administration was followed by decreases in mean arterial pressure, cardiac index, mesenteric, renal and femoral blood flows (MBF, RBF and FBF), and increases in systemic and pulmonary vascular resistances. Fluid resuscitation restored cardiac index, systemic vascular resistance, pulmonary vascular resistance, MBF, RBF and FBF to pre-endotoxin levels. In the presence of endotoxin, ODQ administration increased MBF and prevented the restoration of FBF. Hence, selective inhibition of sGC may increase splanchnic blood flow in septic shock. PMID- 14560111 TI - Mesenteric lymph from rats with thermal injury prolongs the action potential and increases Ca2+ transient in rat ventricular myocytes. AB - Although gut-derived mesenteric lymph from animals with thermal injury appears to lead to myocardial contractile dysfunction, the cellular mechanisms remain unclear. We examined the direct effects of intestinal lymph on excitation contraction coupling in rat ventricular myocytes. Lymph from rats receiving burn injury (burn lymph), but not from sham-burned rats, rapidly enhanced myocyte contraction and the amplitude of Ca2+ transient; the average percentage of shortening was increased from 5.5 +/- 0.3% to 10.5 +/- 0.9%. 90% and the Ca2+ transients increased by 80% +/- 20%. Burn lymph had no effect on the amplitude of L-type Ca2+ current (ICa) or the inward rectifier K+ current, but the transient outward K+ currents (Ito) were reduced significantly by burn lymph. Inhibition of Ito was not altered by an alpha1-adrenergic receptor (AR) antagonist, prazosin, indicating that the block was not mediated via alpha1-AR signaling pathway. Action potential (AP) duration, measured at 50% and 90% repolarization, was prolonged by burn lymph. Stimulation of myocytes with AP voltage-clamp waveforms derived from prolonged AP induced by burn lymph revealed a 1.7-fold increase in Ca2+ influx via ICa compared with the Ca2+ influx induced by control AP. Blocking of Ito by 4-aminopyridine prolonged AP duration and increased Ca2+ transients, mimicking the effects of burn lymph. Burn lymph did not affect Na+/Ca2+ exchange currents or caffeine-induced SR Ca2+ release. Thus, acute exposure of normal cardiac myocytes to burn lymph increases Ca2+ transients by a prolongation of AP as a result of a reduction of Ito with no intrinsic change in ICa or exchanger. The electrophysiological changes are similar to those that occur during compensated cardiac hypertrophy, suggesting a common mechanistic link between burn lymph- and hypertrophy-induced cardiac dysfunction. PMID- 14560113 TI - Delayed administration of alpha-melanocyte-stimulating hormone or combined therapy with BAY 11-7085 protects against gut ischemia-reperfusion injury. AB - Gut ischemia-reperfusion (I/R) injury is a serious complication of shock. Previously we demonstrated that the administration of alpha-melanocyte stimulating hormone (MSH) immediately before mesenteric I/R protected against postischemic gut injury. In this report, we tested the therapeutic efficacy of alpha-MSH on gut I/R (60 min ischemia, 6 h reperfusion) injury when given at different time points of reperfusion. Rats underwent sham surgery or were treated with saline or with alpha-MSH that was given 1, 2, or 4 h after superior mesenteric artery clamping. Vehicle-treated I/R rats exhibited severe mucosal injury and increased NF-kappaB DNA binding activity, myeloperoxidase (MPO) activity, and interleukin-6 and heme oxygenase-1 (HO-1) expression. In contrast, rats given alpha-MSH at 1 h of reperfusion, but not 2 h or 4 h, exhibited much less mucosal injury. Rats given alpha-MSH at 1 h or 2 h of reperfusion, but not 4 h, exhibited less MPO activity, NF-kappaB DNA binding activity, and interleukin-6 protein and even higher levels of heme oxygenase-1 than vehicle-treated rats. In addition, we found that combined use of alpha-MSH, a known inhibitor of IkappaBalpha tyrosine phosphorylation, with BAY 11-7085, an inhibitor of IkappaBalpha Ser 32,36 phosphorylation, abrogates gut MPO induction and tissue injury at early and late time points of reperfusion. Thus, alpha-MSH, an endogenous peptide with a favorable side-effect profile, is effective in treating experimental gut I/R injury when given early after the initial ischemia and may represent a candidate therapy for gut I/R in humans in whom recognition and treatment are often delayed. PMID- 14560114 TI - Splanchnic perfusion during delayed, hypotensive, or aggressive fluid resuscitation from uncontrolled hemorrhage. AB - The purpose of this study was to determine the effect of three different fluid resuscitation strategies on splanchnic perfusion in a clinically relevant model of uncontrolled hemorrhage after liver trauma. Anesthetized swine were instrumented with a gastric near-infrared spectroscopy probe (GStO2), a jejunal tonometer (PrCO2), a portal vein catheter (SpvO2, lactate), and an ultrasonic blood flow probe on the superior mesenteric artery. The liver was lacerated to produce uncontrolled hemorrhage and a shock state characterized by a 40-60% decrease in cardiac output and a decrease in mean arterial pressure (MAP) to 42 +/- 1 mmHg. Animals were randomly assigned to either delayed resuscitation (n = 6); hypotensive resuscitation with lactated Ringer's infusion to MAP = 60 mmHg (n = 6); or aggressive resuscitation with LR to MAP >/= 75 mmHg (n = 6). For the remainder of the protocol, the treatment was identical. The data showed that blood loss (47 +/- 7 and 45 +/- 10 mL/kg) and total fluid requirements (118 +/- 73 and 171 +/- 85 mL/kg) were similar with either hypotensive or aggressive resuscitation. In contrast, with delayed resuscitation, both values were lower (27 +/- 2 mL/kg and 87 +/- 33 mL/kg, both P < 0.05). Despite aggressive resuscitation, SpvO2 and GstO2 were about 10% lower (both P < 0.05 within group) and PrCO2 was about 20 mmHg higher (P < 0.05 within group) than the corresponding values in the other two groups. Thus, delayed resuscitation minimized the blood loss but did not restore tissue oxygenation, whereas aggressive resuscitation was associated with maximal blood loss and splanchnic hypoperfusion. For this reason, it is reasonable to conclude that hypotensive resuscitation might be an effective strategy to maintain splanchnic perfusion after blunt abdominal trauma and uncontrolled hemorrhage. PMID- 14560115 TI - Let every eye negotiate for itself and trust no agent. PMID- 14560116 TI - Limitations of the Panoramic 200 Optomap. PMID- 14560117 TI - Multiple evanescent white dot syndrome and acute zonal occult outer retinopathies. AB - BACKGROUND: Multiple evanescent white dot syndrome (MEWDS) is a rare, acute, multifocal retinitis located in the posterior pole. Patients with MEWDS may report photopsia, central scotoma, or decreased vision. Prodromal viral-like symptoms may be associated with this syndrome, but its etiology remains unknown. The condition is self limiting with a favorable prognosis for visual recovery within several weeks. CASE REPORT: A young, myopic, female patient presented with painless photopsia, scotoma, and subsequent vision loss secondary to MEWDS. The clinical appearance, differential diagnosis, and management of MEWDS are discussed. DISCUSSION: Careful ophthalmoscopic and angiographic interpretation along with complete history taking are the keys to diagnosing MEWDS. Ancillary testing including electroretinogram, early receptor potential, and visual fields aid in the differential diagnosis. Once the diagnosis has been established, patient reassurance is appropriate in the management of this condition. PMID- 14560118 TI - Early use of corrective lenses in Spanish colonies of the Americas including parts of the future United States: reference to Viceroy Luis de Velasco (the Son). AB - We discuss many aspects of a reproduction of a formal painting from the XVIth century. It is a portrait of Viceroy of New Spain Luis de Velasco, El Joven or Hijo (son-see text), found at the Museum of the Pecos National Monument (near Pecos and Santa Fe, New Mexico). A formal portrait of each Viceroy of New Spain was created, and this one was painted during de Velasco's first term as Viceroy (1590 to 1595 AD). In this depiction, he is seen wearing spectacles. To our knowledge, this is the first known reference suggesting the introduction of this form of visual correction in the Spanish Colonies in the Americas and in the future United States. There are three known portraits of Luis de Velasco (son): One painting, which included his father and significant others, hangs in the Colegiata (a small cathedral) de San Luis near Valladolid, Spain. In this portrayal, he was shown as a young man without a spectacle correction. And there are two viceregal portraits (Term 1, 1590 to 1595; Term 2, 1607 to 1611) in which he wears the same spectacle correction (these are located in Mexico City). PMID- 14560119 TI - Change to corneal morphology after refractive surgery (myopic laser in situ keratomileusis) as viewed with a confocal microscope. AB - PURPOSE: This study aimed to look at morphological changes induced by myopic laser in situ keratomileusis (LASIK) in the human cornea using the confocal microscope and to investigate the link between these changes and alterations to corneal sensitivity. METHODS: An in vivo slit-scanning real-time confocal microscope (Tomey ConfoScan P4, Erlangen, Germany) fitted with an Achroplan 40x/0.75 NA immersion objective and a Cochet-Bonnet esthesiometer were used to examine the morphology and sensitivity of the central corneas of six subjects (12 eyes) at an initial visit (before surgery), and at 1 week, 1 month, 3 months, and 6 months after LASIK for myopia. RESULTS: Keratocyte density anterior to the flap interface showed differences between visits (p < 0.0001) and was found to be lower than at the initial visit at 1 week, 1 month, 3 months, and 6 months. Microfolds were noted at the level of the anterior limiting membrane in 11 of 12 eyes after surgery at all visits. Highly reflective flap interface particles were seen in all eyes at all visits after surgery. The subepithelial nerve fiber layer was clearly visible before surgery but could not be imaged in any of the eyes after surgery. Short, unconnected nerve fibers were observed 3 months after surgery; these appeared to form anastomosing interconnections after 6 months. Postsurgical corneal sensitivity was reduced during the first 3 months and recovered to presurgical levels after 6 months. CONCLUSION: LASIK showed a decrease in anterior keratocyte density and microfolds in the anterior limiting membrane, and reflective particles were observed at the flap interface. Corneal sensitivity was depressed during the first 6 months after LASIK surgery; this time course paralleled the appearance of nerve regeneration during this period. Confocal microscopy is capable of providing interesting new insights into the effects of refractive surgery on corneal morphology. PMID- 14560120 TI - The influence of stimulus size on measured visual field extent in infants. AB - PURPOSE: To compare measured visual field extent for a 6 degrees stimulus (typical size used in studies of infants) with a 1.5 degrees stimulus (similar to the largest size used in Goldmann perimetry) in young infants. METHODS: A total of 120 infants (60 each at 3.5 months and 7 months of age) and 24 adults were tested monocularly with a kinetic perimetry procedure using a black double-arc perimeter. Each subject was tested with either a 6 degrees or 1.5 degrees white sphere, which was mounted on a black wand and moved smoothly toward the intersection of the perimeter arms at 3.4 degrees /s. Visual field extent along each perimeter arm was defined as the median of 2 to 3 measurements of the position of the leading edge of the stimulus when the subject made an eye movement toward the stimulus. RESULTS: The 6 degrees stimulus produced larger measured visual field extent than the 1.5 degrees stimulus in 3.5-month olds (temporal field only) and in 7-month olds (nasal and temporal field), but not in adults. CONCLUSIONS: Using the testing conditions of the present study, increasing stimulus size beyond the largest used in a Goldmann perimeter (approximately 2 degrees) increases measured visual field extent in young infants, but not in adults. This may relate to differences in peripheral summation areas or to differences in attentional factors between infants and adults. PMID- 14560122 TI - The theory of object and image eccentricities: a new dimension in ophthalmic optics. AB - During off-axis viewing through an anisometropic spectacle correction, the two eyes must diverge to fuse. This can cause great discomfort, especially in the vertical meridian. To consider a correction for the problem, one must know the amount of forced divergence induced by the spectacles. Because the eyes select ray entry points that do not coincide with the object point projections on the lens planes, the exact directions of gaze are difficult to determine with conventional ray-tracing methods. The methods previously developed for determining the visual directions of gaze through anisometropic spectacle corrections are limited to hypothetical thin lenses and require complicated trigonometric constructs that are not suitable for clinical work. A recently developed method for solving the problem applies the dynamic spectacle magnification to find the visual direction of an image of a given object point. The method, referred to as the theory of object and image eccentricities, is based on projections of object and image from the rotation center onto a common plane, such as the back vertex plane. The theory can be applied to many situations in ophthalmic optics previously difficult to analyze. It does not require ray tracing, and because it considers base curves and thickness, it is more accurate than previous methods. A major advantage is that it considers the two eyes as an integrated system rather than separately and in isolation from each other. PMID- 14560121 TI - Durability of coated CR-39 industrial lenses. AB - PURPOSE: To study the effect of coatings on the resistance of CR-39 industrial plano lenses to ballistic impacts and abrasion from fine particles. METHODS: Twelve groups of CR-39 lenses with various scratch-resistant (SR) or combinations of scratch-resistant and antireflective (SR-AR) coatings were mounted in metal industrial spectacle frames. The ZEST protocol was used to determine the mean impact speed for breakage of each lens group using the Canadian Standards Association ballistic test protocol. One pair of lenses from each group was tested for abrasion resistance using the falling sand method. Abrasion resistance was ranked by the degree of haze observed by three independent observers. RESULTS: Uncoated lenses had the best impact resistance and worst abrasion resistance. SR-coated lenses showed mild to moderate reductions in impact resistance, with no correlation between impact and abrasion resistance. SR-AR coated lenses had very good abrasion resistance, but severely reduced impact resistance. CONCLUSIONS: Most SR-coated CR-39 lenses have a high probability of meeting the high-velocity impact resistance requirement of industrial lenses, whereas CR-39 lenses with SR-AR coats are too fragile to be used in industrial spectacles. As a group, the SR-AR coating tended to be more resistant to abrasion by fine particles and less resistant to ballistic impacts, but the abrasion resistance of the SR-coated lenses was more variable, and, thus, overall there was no significant correlation between impact resistance and abrasion resistance. PMID- 14560123 TI - Learn the truth about medical rumors. PMID- 14560124 TI - Differences in hospice referrals. PMID- 14560125 TI - Praise for emergency contraception info. PMID- 14560126 TI - NP insurance rates likely to increase. PMID- 14560127 TI - The clinical significance of HPV. PMID- 14560128 TI - Treat DVT with low molecular weight heparin. PMID- 14560129 TI - A day in the lives of APNs in the U.S. PMID- 14560130 TI - TNF-alpha inhibitors offer hope to RA patients. PMID- 14560131 TI - New strategy improves lung cancer management. PMID- 14560132 TI - Beware the lost chance theory. PMID- 14560133 TI - Partial nephrectomy: the standard approach for small renal cell carcinoma? PMID- 14560135 TI - Renal cell carcinoma: modern surgical approach. AB - PURPOSE OF REVIEW: In the past decade, minimally invasive therapy options for renal cell carcinoma have been devised in an attempt to minimize operative morbidity while achieving comparable oncologic and functional outcomes. Herein, we evaluate the new developments related to the modern surgical and energy ablative techniques for renal cell carcinoma. RECENT FINDINGS: When compared with the open counterpart, laparoscopic radical and partial nephrectomies have equivalent operative time, decreased blood loss, superior recovery, and improved cosmesis. Nowadays, laparoscopic radical nephrectomy can be performed for pT2 tumors (up to 15 cm), and level I renal vein thrombus is not a formal contraindication for the laparoscopic procedure. Ongoing advances in laparoscopic techniques and operator skills have allowed the development of a reliable technique of laparoscopic partial nephrectomy, which includes the ability to achieve effective intracorporeal renal hypothermia. Cryoablation and radiofrequency ablation therapies have been performed through a laparoscopic or percutaneous approach, using a combination of fine probes and high-resolution imaging studies to precisely target the lesions and accurately monitor the freezing or heating ablation process. Noninvasive tumor ablation can now be achieved by extracorporeally induced high-intensity focused ultrasound. SUMMARY: These minimally invasive techniques represent the modern surgical approach for renal cell carcinoma, aiming to decrease patient morbidity. Laparoscopic radical and partial nephrectomy techniques duplicate the open approach. Results obtained with energy ablative techniques are encouraging. Based on the known slow growth rates of small renal cell carcinoma, one should be cautious when interpreting the short-term results of energy ablative therapies monitored by imaging only. PMID- 14560134 TI - Renal cell carcinoma: new prognostic factors? AB - PURPOSE OF REVIEW: There are limited independent predictors of survival in patients with renal cell carcinoma. Factors related to the tumor, host and treatment may help us to predict prognosis to a certain extent. Prognostic indicators would enable selection of patients who can benefit from adjuvant therapy and thus should be enrolled in clinical trials. This review highlights developments in the identification of current prognosticators for patients with renal cell carcinoma. RECENT FINDINGS: Tumor stage, grade and patient-performance status are the known prognostic indicators in renal cell carcinoma. Besides these parameters, many molecular and cytogenetic markers were evaluated recently. Unfortunately, none of these parameters appear to be a better predictive prognostic factor than the usual staging and grading. Therefore, efforts to identify new markers for tumor proliferation and progression are still ongoing. It was recently reported that low carbonic anhydrase 9 staining may be an independent poor prognostic factor in patients with renal cell carcinoma. Moreover, there is increasing interest in prognostic indices and predictive algorithms for survival. Staging systems that combine the pathological features with additional prognostic variables have been constructed to predict outcome. The UCLA Integrated Staging System seems to be superior to staging alone in differentiating patients' survival. SUMMARY: Although the literature reviewed contains numerous promising clinical, histological, molecular and cytogenetic parameters, none of them has yet been shown to have an independent prognostic value PMID- 14560136 TI - Renal cell carcinoma: relevance of pathology. AB - PURPOSE OF REVIEW: Although pathological findings including tumour grade and tumour-node-metastasis stage provide prognostic information, the outcome for patients with renal cell carcinoma is occasionally unpredictable. Many studies have been undertaken to improve prediction of the prognosis of renal cell carcinoma, along with efforts to clarify the pathogenesis of renal cell carcinoma at the molecular level. This article reviews the advances in renal cell carcinoma research relevant to pathology published between 1 May 2002 and 30 April 2003. RECENT FINDINGS: The establishment of a strict tumour grading system or adding microscopic venous invasion or tumour necrosis is potentially useful for predicting patient outcomes. The contribution of the von Hippel-Lindau-related pathway in early carcinogenesis has been partly revealed with regard to angiogenesis and cell cycle control. Tumour development in renal cell carcinoma can be considered as accumulating heterogenous molecular events related to cytokine production, cell cycle control, anti-apoptotic signal machinery and angiogenesis. SUMMARY: Finding the target genes related to the von Hippel-Lindau pathway may explain the resistance of chemotherapy, and also provide a clue to finding a novel therapeutic option. Molecular targeting therapy can be applicable after more general molecules leading to tumour development and the metastasis of renal cell carcinoma have been identified. PMID- 14560137 TI - Renal cell carcinoma: novel treatments for advanced disease. AB - PURPOSE OF REVIEW: IL-2 or IFN-alpha induce remissions and prolong life in patients carefully selected for a possibly toxic treatment. However, there is a need for better-tolerated and more effective therapies, especially in patients with co-morbidities and those resistant to systemic immunotherapy. Recent achievements in the treatment of advanced renal cell carcinoma highlight potentially significant improvements. RECENT FINDINGS: Cytoreductive surgery or radiation of metastases seems beneficial in well-selected patients, especially as immunotherapy is available. Immune cells within the tumour correlate with response and survival, indicating the importance of local immune modulation. Such modulation has allowed the introduction of well-tolerated treatments such as the inhalation of IL-2 to control lung metastases, which results in a significant survival benefit for high-risk patients, as suggested by a recent outcome study in 200 patients. Antibody-based tumour targeting against cG250, specifically expressed on renal cell carcinoma, seems to stabilize progressive metastatic disease and does not induce toxicity. Vaccination strategies are also well tolerated, but have not yet shown convincing results in advanced disease. Other approaches have not fulfilled expectations. Thalidomide has significant neurotoxicity and its efficacy was not confirmed in recent studies. Stem cell transplantation has significant toxicity, and cannot yet be recommended, but may have future potential. SUMMARY: Cytokine-based immunotherapy can now be considered standard in the treatment of metastatic renal cell carcinoma. There is good evidence that additional local procedures such as surgery, radiation or the inhalation of IL-2 improve response and survival in metastatic disease with moderate toxicity, resulting in a significant improvement for patients suitable for these approaches. PMID- 14560138 TI - Basic science and research in renal cell carcinoma: from workbench to bedside. AB - PURPOSE OF REVIEW: Renal cell carcinoma represents the third most common cancer in men. Radical surgery remains the only curative approach, and the 5-year survival rate once the cancer has metastasized rarely exceeds 20% despite systemic therapy. It becomes evident that an improvement in outcome might only be achieved if (1) there is early diagnosis, (2) there is accurate prediction of progression and response, and (3) new treatment options reflecting the molecular pathogenesis and progression are developed. RECENT FINDINGS: The detection of circulating cancer cells by reverse transcriptase/polymerase chain reaction techniques for the MN/CAIX gene, the identification of specific genetic alterations in circulating tumor DNA, as well as the demonstration of somatic von Hippel-Lindau mutations and extracellular matrix proteins in urine of high-risk patients might be clinically useful in improving early diagnosis and treatment. The signal transducer and activator of transcription has been shown to significantly correlate with relapse patterns following radical surgery. Heterozygosity or homozygosity for class II haplotypes DQA1 and DQB1 accurately predicts response and survival following cytokine-based therapy and may be helpful in patient selection. In terms of treatment, the use of monoclonal antibody derivates against the epidermal growth factor receptor and the vascular endothelial growth factor receptor has shown promising clinical results. Antisense oligodeoxynucleotide therapy has shown significant therapeutic effects in in-vitro and in-vivo studies. Recent developments in the clinical application of proteasome inhibitors have opened the door to exciting, highly specific and effective molecular treatment options for metastatic renal cell carcinoma. SUMMARY: Recent developments in research on renal cell carcinoma have identified various clinically useful diagnostic and therapeutic options reflecting the molecular basis of the pathogenesis and progression of the disease. PMID- 14560139 TI - Genetics in renal cell carcinoma. AB - PURPOSE OF REVIEW: The combination of several recent molecular technologies, including comparative genomic hybridization, fluorescence in-situ hybridization and complementary DNA and tissue microarrays, has advanced our understanding of renal cancer. However, a great deal of information regarding the genetics of renal neoplasms has also emerged from the extensive cytogenetic investigations in the past decade. RECENT FINDINGS: The correlation between cytogenetic or molecular genetic abnormalities and histomorphology is most consistent in clear cell and papillary types of renal cell carcinoma. However, gene expression profile studies have brought new insights into the classification of renal tumors, and may provide new markers that identify patients with a poor prognosis as well as identifying potential therapeutic targets. SUMMARY: The integration of expression profile data and clinical parameters could serve to enhance the diagnosis and prognosis of renal cell carcinoma. The identification and evaluation of new molecular parameters will be necessities in cancer research and cancer treatment. PMID- 14560140 TI - Penile cancer: review of the recent literature. AB - PURPOSE OF REVIEW: To review recent developments in the diagnosis and treatment of penile cancer, we reviewed the medical literature from 1 May 2002 to 1 June 2003 on the incidence, aetiology, staging, diagnosis and treatment of penile cancer by a Medline search. RECENT FINDINGS: The diagnostic tools and treatment modalities for penile cancer are still controversial. Recently developed diagnostic and therapeutic modalities should help urologists in the management of the disease. SUMMARY: Our review of the recent literature provides an update on diagnostic and therapeutic opinion on penile cancer. PMID- 14560141 TI - Stage I testicular cancer. AB - PURPOSE OF REVIEW: To review current developments in the management of patients with testicular cancer, with special emphasis on risk factors for the primary tumour and treatment options for clinical stage I testicular germ cell tumours. RECENT FINDINGS: The management of patients with testicular cancer has substantially improved over the past 25 years. Current concepts for treating localized and regional disease have been influenced by effective systematic chemotherapy. At present, cure rates approach nearly 100% for low-stage disease and more than 80% for advanced disease. SUMMARY: Retroperitoneal lymph node dissection is still favoured as the therapy of choice for clinical stage I non seminomatous germ cell tumours in many centres, but as risk factors for the primary tumour have become better understood, surveillance and risk-adapted therapy, including surveillance for low-risk patients and adjuvant chemotherapy for the high-risk group, is now being considered a therapeutic option particularly in European centres. Adjuvant radiotherapy is still the gold standard for the treatment of patients with clinical stage I seminoma, but the relapse rate of 19% and a 5-year overall survival of 97.7% make surveillance a possible therapeutic option. The results of phase II and III trials should soon provide additional information on carboplatin for single-agent adjuvant chemotherapy. PMID- 14560142 TI - Bladder substitution. AB - PURPOSE OF REVIEW: Radical cystectomy and urinary diversion is the accepted standard of care for invasive bladder cancer, with orthotopic neobladders as the preferred method of bladder substitution in male and female patients. Last year's studies and reports on this subject are summarized and reviewed. RECENT FINDINGS: Today, most patients considered candidates for radical cystectomy, independent of sex, are potential candidates for orthotopic bladder substitution. It has been shown that orthotopic bladder replacement is well tolerated and feasible even in selected patients with locally advanced tumors with or without positive nodes. The complication rates for orthotopic bladder substitutes are similar to, or lower than, the morbidity rates after conduit formation or continent cutaneous diversion. It has been shown that an afferent tubular segment without any antireflux flap-valve-type implantation techniques protects the upper urinary tract adequately for more than a decade. The true impact of the chosen type of urinary diversion on distress symptoms, quality of life and patients' well-being among survivors of bladder cancer remains controversial and requires further clinical investigation. SUMMARY: The excellent functional outcome and voiding pattern with orthotopic bladder substitution, even in the long-term, argues in favor of this technique as the preferred method for lower urinary tract reconstruction following radical cystectomy for invasive bladder cancer. PMID- 14560143 TI - Flap technology for reconstructions of urogenital organs. AB - PURPOSE OF REVIEW: The purpose of this review is to summarize the different reconstructive options for urogenital indications. The development of various flap techniques to restore congenital and acquired urogenital defects is presented. RECENT FINDINGS: Various reconstructive techniques have been demonstrated recently. On the basis of the reconstructive requirements, two main techniques can be defined: the standard local or regional flap technique (pedicled flap) and the more sophisticated microvascular free flap technique. Free tissue transplantation (transfer) is a procedure that involves microvascular transplantation of a flap (a fasciocutaneous, muscle or composite flap) in one stage from a donor site in the body to a distant recipient site. The viability of the transplanted flap is maintained by microvascular anastomosis between the flap's vessels (at least one artery and one vein) and recipient vessels. Re innervation and functioning muscle contraction is achieved by suturing the vessels and a motor nerve in the recipient area to a motor nerve of a free transplanted muscle. After regeneration of the nerve and re-innervation of the transplanted muscle, a functioning free transplanted muscle offers enough contractile capacity and strength to replace the function of the missing muscles at the recipient site. The technique of microvascular free tissue transfer necessitates extensive experience in microvascular technique and this approach could be efficiently applied in cooperation with other specialists. Recent studies show the development and clinical application of these new surgical techniques in urology (e.g. in the treatment of bladder acontractility using innervated free latissimus dorsi muscle and in the use of a free microvascular fillet lower leg flap for the reconstruction of a large pelvic-floor defect). SUMMARY: Various reconstructive requirements define the techniques for reconstruction. The main principle is to obtain optimal anatomical and functional reconstruction with minimal donor site morbidity. Depending on the etiology of the defect, different reconstructive options are available to optimize the reconstructive result. Optimal reconstruction might best be achieved by adopting an interdisciplinary approach in which the primary objective is to provide the best possible outcome for each patient. This review presents the main indications for and principles of flap selection according to the reconstructive requirements. PMID- 14560144 TI - Vaginal and penile reconstruction. AB - PURPOSE OF REVIEW: Reconstructive surgery for patients with genital abnormalities or for patients who require reconstructive efforts is challenging. This review highlights those articles, which are outstanding among all those important papers, which have been published during the last year (2002-2003). RECENT FINDINGS: A greater understanding of embryonal development improves the success of reconstructive surgery. Other factors, such as the patient's sex, influence the surgical technique used and the degree of invasiveness or complexity. In the adult the pressure to shorten hospital stays has played a big part in the continual modification and enhancement of surgical techniques. In addition to modified techniques, new off-the-shelf materials are introduced to the clinic, which seem to have the potential to improve the surgical outcome and shorten hospital stays. SUMMARY: With the continued successful basic anatomy and basic research, reconstructive surgery brings higher success rates. Long-term results are still required to validate the reliability of these new surgical techniques and materials. PMID- 14560145 TI - Nerve regeneration: might this be the only solution for functional problems of the urinary tract? AB - PURPOSE OF REVIEW: To assess the potential role of nerve regeneration in restoring urinary tract function, the rapidly developing and exciting area of central and peripheral nerve repair and regeneration is reviewed, with particular reference to papers in which animal models of nerve damage resulting in urogenital dysfunction have been used. The difficulties and potential of these techniques for therapeutic application to human subjects with functional problems of the urinary tract are discussed. RECENT FINDINGS: Methods for encouraging regeneration of cut axons and directed growth in the inhibitory environment of the central nervous system are being extensively explored. The recent discovery of the potential of olfactory ensheathing cells has proved a significant advance. Olfactory ensheathing cells are a type of glial cell which can be harvested from the olfactory mucosa. Transplantation of these cells, in conjunction with a biodegradable synthetic nerve guide or conduit, has been shown to restore urinary tract function after spinal cord injury. Artificial, biodegradable conduits have also restored bladder and spermatic duct function after sympathetic nerve damage. Other adjuvants facilitating the process of axonal recovery include the use of neurotrophins to accelerate and guide the formation of new nerve-fibre growth. SUMMARY: These revolutionary technologies may, in the future, provide a means of treating urinary tract dysfunction with some types of aetiology, including acute spinal cord injury, and injury to nerves following pelvic surgery. It is, however, less likely that these treatments will be used successfully in the near future in patients in which the neural damage is long term, or associated with death of post-ganglionic neurons. PMID- 14560146 TI - High-grade dysplasia in sporadic fundic gland polyps: clinically relevant or not? AB - Sporadic fundic gland polyps (FGPs) are the most frequent gastric polyps. These FGPs develop almost exclusively in a normal corpus mucosa without Helicobacter pylori infection. This means that they represent an absolutely benign disease (no ulcers, almost no gastric carcinoma and mucosa associated lymphoid tissue (MALT) lymphoma). Most probably there exists a statistically accumulated coincidence rate of sporadic FGPs with colorectal epithelial neoplasias, so it is recommended to consider preventive colonoscopy. In familial adenomatous polyposis (FAP), FGPs are present in up to 80-93% of patients. In such patients, low-grade dysplasia and/or foveolar microadenomas may be present in 44-54% of cases. To date, only three case reports on early gastric carcinoma have been published. In this issue, to the best of our knowledge the first case of a high-grade dysplasia (intraepithelial neoplasia) in a sporadic FGP is reported. Recently, it has been shown that beta-catenin mutations might have led to FGP. Therefore, in the future, patients showing nuclear beta-catenin expression must undergo examination to discover whether this expression may lead to neoplasia. According to the data available at present, such a constellation must be regarded as an extremely rare event. PMID- 14560147 TI - High prevalence of combined thrombophilic abnormalities in patients with inflammatory bowel disease. AB - INTRODUCTION: A hypercoagulable state has been recognized in patients with inflammatory bowel disease. OBJECTIVE: The aim of this study was to determine the frequency of single and combined thrombophilic abnormalities in patients from northern Portugal with Crohn's disease or ulcerative colitis, without a history of thrombosis. METHODS A cross-sectional study involving 116 patients (42 with ulcerative colitis, 74 with Crohn's disease), and 141 randomly chosen asymptomatic blood donors was carried out. Prothrombotic variables and genetic abnormalities were assessed. RESULTS: The prevalence of single prothrombotic abnormalities (only one alteration) in inflammatory bowel disease patients was higher than in the reference population (26% and 18%, respectively; P < 0.02). The allelic frequency of genetic polymorphisms was higher in Crohn's disease and ulcerative colitis for MTHFR C677T, ACE Del and PAI-1 4G (P < 0.001) than in the reference population. The prevalence of combined thrombophilic abnormalities (at least two alterations) in both Crohn's disease and ulcerative colitis was also higher (22% and 21%, respectively) than in the reference population (9%; P < 0.01). These differences were not related to age or gender; however, in Crohn's disease the frequency of two or more abnormalities was related to disease activity (odds ratio 3.0 [1.3-6.7]). CONCLUSION: Higher prevalences of single and combined thrombophilic defects were found in inflammatory bowel disease patients, factors that could be involved in the disease pathogenesis. PMID- 14560148 TI - Effect of systemic glucocorticoid therapy on bone metabolism and the osteoprotegerin system in patients with active Crohn's disease. AB - BACKGROUND AND AIMS: Osteoporosis may occur in 25-30% of patients with Crohn's disease. Its pathogenesis is not completely understood. Both systemic inflammation in acute disease and treatment with systemic glucocorticoids have been implicated. The aim of the present study was to investigate changes in bone density and biochemical markers of bone metabolism before and during a 3-month period of high-dose glucocorticoid treatment for acute flare-up of Crohn's disease. METHODS: Twenty-five patients with active Crohn's disease requiring systemic glucocorticoid treatment (prednisolone, 60 mg/day) were investigated. Lumbar spine and femoral neck bone mineral densitometry was performed at baseline and again after 3 months. Clinical examinations including evaluation of the Crohn's disease activity index and measurement of the biochemical markers osteocalcin, deoxypyridinoline, osteoprotegerin and the soluble receptor activator of NF-kappaB ligand were performed prior to, and at 1, 2 and 12 weeks following steroid administration. RESULTS Median lumbar bone mineral density decreased significantly during the observation period by 1.04% from -0.84 (t score; range, -2.8 to +0.57) to -0.95 (range, -3.1 to +0.40; P = 0.022), while bone density of the total femur decreased by 2.9% from -0.83 (range, -2.61 to +1.86) to -0.90 (range, -2.65 to +0.19; P = 0.01). Serum levels of osteocalcin, a bone formation marker, and osteoprotegerin, an anti-resorptive cytokine produced by osteoblasts, decreased after the first 2 weeks of treatment and reached baseline levels after 3 months. No significant change was found for the bone resorption marker deoxypyridinoline, while soluble receptor activator of NF kappaB ligand, a cytokine promoting bone resorption, tended to increase during steroid treatment. CONCLUSION: A decrease in bone mineral density in patients with Crohn's disease appears to result, at least in part, from a short-term effect of systemic glucocorticoid. Modulation of osteoclastogenesis by the receptor activator of NF-kappaB ligand/osteoprotegerin cytokine system and decreased osteoblastic function may be the underlying molecular basis. PMID- 14560149 TI - Detection of identical Helicobacter DNA in the stomach and in the non-cirrhotic liver of patients with hepatocellular carcinoma. AB - OBJECTIVES: Studies have implied the presence of Helicobacter species in the human liver. The possible relationship between the presence of Helicobacter species in the non-cirrhotic liver of patients with hepatocellular carcinoma (HCC) and the detection of Helicobacter species in their stomach was explored. PATIENTS AND METHODS: A 16S rDNA-based polymerase chain reaction (PCR) followed by DNA sequence analysis of the obtained PCR fragments was performed on 51 surgically obtained non-cirrhotic liver specimens and 14 gastric samples. RESULTS: Analysis indicated a significant difference in the presence of Helicobacter-species-specific DNA in the liver of patients with HCC compared with controls. Sequence analysis of these PCR products obtained from HCC patients indicated that they were related most closely to the 16S rDNA sequence of Helicobacter pylori but that they always differed at the same two positions. This same aberrant Helicobacter-species-specific 16S rDNA could be isolated in gastric samples of patients with HCC. CONCLUSION: These data suggest that gastric colonization with a specific subset of Helicobacter strains is associated with the induction of HCC, either directly via colonization of the liver or indirectly, e.g. via secretion of specific toxins by Helicobacter residing in the stomach. PMID- 14560150 TI - Quality of life in patients with functional dyspepsia: a prospective 1-year follow-up study in Spanish patients. AB - BACKGROUND AND AIMS: The natural history of functional dyspepsia is not well known. We prospectively assess the quality of life and severity of symptoms in a group of Spanish patients with functional dyspepsia. PATIENTS AND METHODS: One hundred and twelve consecutive patients with functional dyspepsia, according to Rome II criteria, were prospectively followed up for 1 year. All patients completed symptom (Dyspepsia Questionnaire and the Gastrointestinal Symptoms Rating Scale) and quality of life [the Psychological General Well-Being (PGWB) Index and the General Health Questionnaire (GHQ)] questionnaires every 3 months. Only free antacid consumption was permitted during the study period. RESULTS: The group was made up of 81 women and 31 men with a mean age of 45 +/- 17 years; 66% of patients were infected with Helicobacter pylori, and ulcer-like dyspepsia (53%) was the predominant subgroup. At baseline, quality of life scores were low (PGWB, 87.1 +/- 17.6 and GHQ, 20.6 +/- 11.8), but these values gradually improved during the year of follow-up (PGWB, 107.7 +/- 1.1 and GHQ, 8.9 +/- 0.4). Digestive symptoms also decreased. In the multivariate analysis, the anxiety score on the PGWB index (Wald, 5.2; P = 0.02) and smoking status (Wald, 4.3; P = 0.04) were predictors of end quality of life. At baseline, patients with a high level of anxiety had a very reduced quality of life, although their symptom scores were similar to other patients. CONCLUSION: Quality of life is reduced in patients with functional dyspepsia. Some improvement in quality of life together with a decrease in the severity of symptom scores was seen during the 1 year of follow-up. We believe that both the reassurance of negative endoscopy and the scheduling of visits to the doctor favourably influence the quality of life. PMID- 14560151 TI - Genotypic distribution of hepatitis B virus in the Czech Republic: a possible association with modes of transmission and clinical outcome. AB - OBJECTIVE: The relationship between hepatitis B virus (HBV) genotype with disease or treatment outcome is beginning to be characterized. However, the link between genotype and disease transmission route has not been closely examined. We addressed this question in high-risk populations in Prague, Czech Republic. DESIGN: Patients with HBV infection were consecutively recruited into the study at an outpatient clinic between June 2000 and March 2001. Their serum samples were analysed for HBV S gene segments by polymerase chain reaction (PCR) test. The amplified product sequences were compared to those of known HBV genotypes. Patients were evaluated for other virus co-infections, and parenteral and sexual exposure histories. RESULTS: Of 57 consecutively recruited patients with evidence of HBV infection, 45 (79%) had PCR-detectable S gene sequences. Only genotypes A (n = 33; 73%) and D (n = 12; 27%) were found. There was no difference in the development of chronic infection between the two genotypes. Of nine patients co infected with TTV, all were infected with HBV genotype A. There was a trend towards an association between number of lifetime sex partners and genotype A but not genotype D. CONCLUSIONS: In Prague, the number of HBV genotypes appears to be limited compared to other northern European countries, suggesting that the virus has recently spread in the high-risk populations. While a large proportion of HBV infections occur in intravenous drug users, a subset of HBV genotype A may be transmitted by sexual contact. An HBV subtype may influence modes of transmission of HBV. PMID- 14560152 TI - Endoscopic ultrasonography-guided fine needle aspiration biopsy in follow-up patients with gastrointestinal stromal tumours. AB - OBJECTIVES: Since all gastrointestinal stromal tumours (GISTs) are practically considered to be potentially malignant, they must be clinically differentiated from other submucosal tumours (SMTs). In this study, we carried out endoscopic ultrasonography-guided fine needle aspiration biopsy (EUS-FNAB) against follow-up SMTs to examine whether the method is sufficient for pathological diagnosis and analysis of c-kit mutation. The relationship between tumour growth and c-kit mutation was also examined. METHODS: The tumour tissues were obtained by EUS-FNAB from 10 Japanese follow-up cases with SMT. Paraffin sections of tissues were used for haematoxylin and eosin staining, and for immunohistochemistry. Genomic DNA was extracted from the sections, and c-kit gene fragments of exons 9, 11, 13 and 17 were amplified by polymerase chain reaction and directly sequenced. RESULTS: Nine SMTs were diagnosed as GIST and one was diagnosed as schwannoma by immunohistochemistry. Mutation at exon 11 was detected in six of nine GISTs, but no mutations were detected at exons 9, 13 and 17 in all GISTs. The case of schwannoma did not have any mutations at exons 9, 11, 13 and 17. Statistical significance was not observed between the average growth rate of six GISTs with the mutation and that of three GISTs without the mutation (P = 0.694). CONCLUSIONS: EUS-FNAB against SMTs was useful for the differential diagnosis of SMT and the analysis of c-kit mutation. The c-kit mutation might not be one of the predictable markers for rapid tumour growth. PMID- 14560153 TI - Cellular turnover of normal gastrointestinal epithelium assessed by changes in telomeric: total DNA signal ratios. AB - OBJECTIVES: The reasons why different areas of gastrointestinal mucosa exhibit widely different rates of malignant change are still poorly understood. Malignancy rates rise markedly with age. We therefore hypothesised that rates of malignant change might correlate with rates of ageing as judged by stem cell turnover. Telomeric DNA is lost with each cell division and so acts as a measure of the number of cell divisions undergone by stem cells. We measured telomeric:total DNA signal ratios in normal gastric (Helicobacter pylori-positive and H. pylori-negative), duodenal and colonic mucosa to see whether ratios correlated with propensity to malignancy. PATIENTS: Subjects undergoing diagnostic upper (n = 93) or lower (n = 45) gastrointestinal endoscopy, whose mucosa appeared macroscopically normal, sampled over a wide age range. METHODS: DNA was extracted from paired blood and mucosal samples (colonic or gastric and duodenal). Telomere length was assessed by dot blot hybridisation with an oligonucleotide-containing telomeric sequence compared with the signal obtained from total genomic DNA. Helicobacter status was assessed by Campylobacter-like organism (CLO) test and serologically. RESULTS: Telomeric signal ratios were scattered, but correlated within individuals. The ratios tended to decrease with age but the rates of decrease did not correlate with rates of malignant change. Gastric tissue had the shortest ratios and duodenal ratios decreased fastest. CONCLUSIONS: The telomeric signal ratios did not suggest any obvious basis for differential rates of disease especially malignancy. Infection with H. pylori was not associated with lower gastric telomere ratios. PMID- 14560154 TI - Diabetes mellitus is a recurrence-independent risk factor in patients with hepatitis B virus-related hepatocellular carcinoma undergoing resection. AB - OBJECTIVES: The prognostic impact of diabetes mellitus and its association with virological factors on patients with hepatocellular carcinoma (HCC) are unclear. This study aimed to investigate the outcome in HCC patients undergoing resection with and without diabetes mellitus and the interaction with hepatitis B virus (HBV) and hepatitis C virus (HCV). METHODS: A total of 239 HCC patients were included. Survival and tumour recurrence were analysed according to the status of diabetes mellitus and viral hepatitis. RESULTS: Diabetic patients tended to have an overall decreased survival compared with non-diabetic patients in 32 +/- 19 months of follow-up. A total of 113 (47%) patients developed tumour recurrence during the follow-up period. No significant difference of tumour recurrence between diabetic and non-diabetic patients, or between patients seropositive and seronegative for HBV surface antigen (HBsAg), was noted. Stratified analysis showed that diabetic patients with HBV but not with HCV infection had a poor long term outcome. In the HBV group, the one-, three- and five-year survival rates were 83%, 51% and 41% versus 90%, 78% and 73% in diabetic and non-diabetic patients, respectively, compared with 90%, 83% and 42% versus 91%, 73% and 73% in the HCV group with and without diabetes mellitus, respectively. Cox multivariate analysis disclosed that diabetic patients seropositive for HBsAg had a significantly poor survival. CONCLUSIONS: Diabetes mellitus does not affect the long-term survival in HCV-related HCC but is a recurrence-independent poor prognostic factor for HBV-related HCC. PMID- 14560155 TI - Plasma pituitary adenylate cyclase activating polypeptide (PACAP) levels in chronic hepatitis B patients under lamivudine treatment. AB - OBJECTIVE: Lamivudine is a nucleoside analogue with potent antiviral activity against hepatitis B virus (HBV). Plasma pituitary adenylate cyclase activating polypeptide (PACAP) is a multifunctional neuropeptide that is produced within the lymphoid microenvironment and induces the production of Th2-type cytokines. The aim of our study was to investigate the possible alterations of plasma PACAP-38 levels in chronic hepatitis B (CHB) patients during lamivudine treatment and to compare them with biochemical, virological and histological data. METHODS: Plasma PACAP-38 levels were measured using competitive radio-immune analysis (RIA) in 25 CHB patients before and after completion of a 52-week lamivudine treatment period and in 22 healthy blood donors. Biochemical evaluation was done at baseline and every three months during treatment. Virological evaluation (HBV-DNA) was performed at baseline and at weeks 24 and 52 of treatment. Baseline liver histology was assessed for all patients at the beginning and at week 52 of the study for histological comparison with the pretreatment biopsy, according to the Ishak scoring system. Statistical evaluation of data was done using analysis of variance and Student's t-test. RESULTS: Virological breakthrough was observed in seven (28%) patients at week 52 of treatment. Histological improvement was observed in 21 (84%) CHB patients, despite the emergence of tyrosine-methionine aspartate-aspartate (YMDD) mutations. Plasma PACAP-38 levels were significantly lower in CHB patients at baseline than in healthy blood donors. Significant elevation of plasma peptide levels was observed in CHB patients after the completion of lamivudine treatment period, even in the subgroup of those who exhibited YMDD variants. CONCLUSION: The elevation of plasma PACAP-38 levels in treated CHB patients following lamivudine-induced elimination of viraemia suggests a possible alteration of T-cellular immune response, resulting in biochemical and histological remission of liver disease, even in patients who exhibited virological breakthrough. PMID- 14560156 TI - Characteristics and regional variations of patients with Barrett's oesophagus in the UK. AB - OBJECTIVES: To establish the patient characteristics and regional variation of Barrett's oesophagus in the UK in a large number (5717) of subjects. Barrett's oesophagus is a precursor lesion of oesophageal adenocarcinoma, the incidence of which is rising more rapidly than any other solid tumour. METHODS: Demographic data of 5717 Barrett's oesophagus patients from 27 UK centres, each registering at least 50 patients, with UK National Barrett's Oesophagus Registry, were analysed. RESULTS: Centres registering patients were distributed evenly throughout the UK. There was an excess of males, with a male to female ratio of 1.7. Mean age at diagnosis was 62.0 years for males and 67.5 years for females. In Scotland, the mean and peak age at diagnosis for males was lower than for the remainder of the UK. In contrast, there was no regional variation in mean and peak age of diagnosis for females. In 3880 Barrett's oesophagus patients with complete data, adenocarcinoma developed in 136 (3.5%). Adenocarcinoma prevalence was 4.0% in males and 2.5% in females. The male/female ratio of patients with adenocarcinoma was 3.0, twice that of Barrett's oesophagus (1.7). The mean age was 64.7 years for males and 74.0 years for females. CONCLUSIONS: These data demonstrate that Barrett's oesophagus occurs with a higher frequency in males, with progression to adenocarcinoma being even more biased towards men in the UK, in contrast to the USA. There are significant regional variations in age of diagnosis of Barrett's oesophagus, especially in younger male patients in Scotland. Studies to identify genetic and environmental determinants of Barrett's oesophagus and cancer risk are needed. PMID- 14560157 TI - Analysis of tumour necrosis factor alpha and interleukin 10 promotor variants in patients with chronic pancreatitis. AB - OBJECTIVE: Cationic trypsinogen gene mutations are strong risk factors of hereditary pancreatitis. However, 20% of subjects with a trypsinogen mutation never get pancreatitis and the cause of this incomplete penetrance is unknown. We investigated the influence of interleukin 10 (IL10) and tumour necrosis factor alpha (TNFalpha) promotor variants on the manifestation of chronic pancreatitis of different underlying causes and in pancreatic cancer. METHODS: A total of 335 German patients with chronic pancreatitis were investigated. In 157 patients the disease was related to alcohol abuse; the other cases were of non-alcoholic origin. In the latter group, the serine protease inhibitor, Kazal type 1 (SPINK1) mutation N34S was found in 72 patients and the trypsinogen mutations N29I or R122H were present in 60 patients; in the remaining 46 patients no mutation was found. In addition, we studied 208 patients with pancreatic cancer. As controls, 116 healthy blood donors and 25 healthy carriers of the trypsinogen mutations N29I or R122H were investigated. After DNA extraction from blood leucocytes, genotyping for the cytokine polymorphisms was performed by induced heteroduplex generators and/or direct DNA sequencing of the IL10 and TNFalpha promotor regions. RESULTS: The frequencies of the promotor polymorphisms of IL10-627A, IL10-1117A, TNF-238A and TNF-308A in patients with alcoholic chronic pancreatitis, idiopathic pancreatitis, SPINK1-N34S-associated chronic pancreatitis and pancreatic cancer did not differ significantly from the control group. The variant TNF-238A was two to four times more frequent in index patients with trypsinogen mutations than in all other groups. The analysis of the allelic frequencies of whole families with trypsinogen mutations revealed that all subjects with the TNF-238A variant suffered from chronic pancreatitis, whereas all intrafamilial controls with wild-type TNF were unaffected. CONCLUSIONS: TNFalpha and IL10 promotor variants are not associated with a manifestation of chronic pancreatitis or pancreatic cancer. The variant TNF-238A, however, might be a relevant risk factor for disease manifestation in families with hereditary pancreatitis. PMID- 14560158 TI - High-grade dysplasia in sporadic fundic gland polyps: a case report and review of the literature. AB - We present a case of fundic gland polyps (FGPs) containing high-grade dysplasia in a 68-year-old man. High-grade dysplasia, and even gastric adenocarcinoma, associated with FGPs have been described in patients with familial adenomatous polyposis (FAP) and attenuated familial adenomatous polyposis (AFAP) but never in non-FAP patients. Two colonoscopies in the past six years virtually rule out FAP and AFAP in our patient. Dysplasia in FGPs from non-FAP patients is extremely rare, and until now only cases of low-grade dysplasia have been described. The literature on dysplasia in FGPs is reviewed briefly. Additional immunohistochemical investigations in this case showed nuclear staining of beta catenin, increased proliferation and apoptosis in the dysplastic areas of the FGPs. Our case suggests that the malignant potential of FGPs is not limited to FAP-associated FGPs. PMID- 14560159 TI - Metastatic liver disease and fulminant hepatic failure: presentation of a case and review of the literature. AB - Although liver metastases are commonly found in cancer patients, fulminant hepatic failure (FHF) secondary to diffuse liver infiltration is rare. Furthermore, clinical presentation and laboratory findings are obscure and far from being pathognomonic for the disease. We report a case of a patient who died in the intensive care unit of our hospital from multiple organ failure syndrome secondary to FHF, as a result of liver infiltration from poorly differentiated small cell lung carcinoma. We also present the current knowledge about the clinical picture, laboratory findings and physical history of neoplastic liver metastasis-induced FHF. PMID- 14560160 TI - Gastric hepatoid adenocarcinoma and familial investigation: does it always produce alpha-fetoprotein? AB - We present the case of a 68-year-old Caucasian man with gastric hepatoid adenocarcinoma without increased levels of alpha-fetoprotein (AFP) in the serum. The patient had a strong history of gastric cancer in his family, affecting seven members, including a brother and a sister. The patient underwent subtotal gastrectomy, but 4 months later presented hepatic metastases, and 6 months after the initial diagnosis he succumbed to the disease. Immunohistochemical tests showed that the tumour was positive for AFP, hepatocyte paraffin 1, and neuron specific enolase, but negative for synaptophysin and chromogranin. Previously reported cases of hepatoid gastric tumours showed that they produce large amounts of AFP and that they have a poor prognosis. PMID- 14560161 TI - Mistaken identity: misclassification of TPMT phenotype following blood transfusion. AB - Azathioprine (AZA) is an effective treatment for inflammatory bowel disease. The measurement of thiopurine methyltransferase (TPMT) activity levels helps to identify the one in 300 patients who are at risk of profound myelosuppression with standard doses of AZA. Thus, it is important that the measurement of TPMT activity is accurate. We report a case of misclassification of TPMT phenotype caused by prior blood transfusion. The patient appeared to have an intermediate level of TPMT enzyme activity as measured after the blood transfusion. However, following severe myelosuppression soon after starting AZA, her genotype was determined, which showed that she was homozygous for low TPMT activity. This report reviews some of the limitations in determining both the phenotype and the genotype of TPMT, especially following recent blood transfusion. The dilemma of whether or not to wait for the TPMT activity result before starting AZA is also discussed. PMID- 14560163 TI - Depression and suicidality in preschoolers. PMID- 14560164 TI - Late-talking toddlers. PMID- 14560165 TI - Ten-year review of rating scales. VI: scales assessing externalizing behaviors. AB - OBJECTIVE: This is the sixth article in a series of 10-year reviews of rating scales. The current article reviews scales that assess externalizing behaviors such as disruptive behavior disorders and aggression. METHOD: Relevant scales were found by searching popular electronic databases. The search was then broadened by a review of the references in selected articles. Due to the paucity of well-established scales, any such scales with potential utility for elucidating the functioning of youths with externalizing behaviors were selected. RESULTS: None of these scales is diagnosis-based, although some correlate with DSM-IV-defined disruptive behavior disorders. Most scales assessing disruptive behavior disorders have a solid normative base, good psychometric functioning, and high clinical utility. Scales assessing aggression comprise a bimodal group. Several have been adapted from the adult literature and are widely used in clinical practice, while others address theoretical aspects of aggression and are used predominantly in research. Empirical support for all of the scales assessing aggression varies widely, although several show potential for routine clinical practice. CONCLUSIONS: Overall, these scales measure a variety of constructs with considerable utility for assessing youths' externalizing behaviors, predicting outcome, and evaluating treatment effects. Many need further validation with youth. PMID- 14560166 TI - Brief psychoeducational parenting program: an evaluation and 1-year follow-up. AB - OBJECTIVE: Despite recognition of the need for parenting interventions to prevent childhood behavioral problems, few community programs have been evaluated. This report describes the randomized controlled evaluation of a four-session psychoeducational group for parents of preschoolers with behavior problems, delivered in community agencies. METHOD: In 1998, 222 primary caregivers, recruited through community ads, filled out questionnaires on parenting practices and child behavior. Parents were randomly assigned to immediate intervention or a wait-list control. The intervention comprised three weekly group sessions and a 1 month booster, the focus being to support effective discipline (using the video 1 2-3 Magic) and to reduce parent-child conflict. RESULTS: Using an intent-to-treat analysis, repeated-measures analyses of variance indicated that the parents who received the intervention reported significantly greater improvement in parenting practices and a significantly greater reduction in child problem behavior than the control group. The gains in positive parenting behaviors were maintained at 1 year follow-up in a subset of the experimental group. CONCLUSIONS: This brief intervention program may be a useful first intervention for parents of young children with behavior problems, as it seems both acceptable and reasonably effective. PMID- 14560167 TI - Exposure-based cognitive-behavioral treatment for phobic and anxiety disorders: treatment effects and maintenance for Hispanic/Latino relative to European American youths. AB - OBJECTIVE: To examine treatment response and maintenance to exposure-based cognitive-behavioral therapy (CBT) for Hispanic/Latino relative to European American youths with phobic and anxiety disorders. METHOD: A total of 131 Hispanic/Latino and European-American youths (aged 6-16 years) who participated in two previous clinical trials for phobic and anxiety disorders were compared along diagnostic recovery rates, clinically significant improvement, and youth- and parent-completed questionnaire scores using traditional hypothesis tests, including effect sizes, and statistical equivalence tests. RESULTS: After treatment, Hispanic/Latino and European-American youths responded similarly to exposure-based CBT in their diagnostic recovery rates and questionnaires. Effect sizes for questionnaire data were also more similar than different across the two groups. With regard to treatment maintenance, Hispanic/Latino and European American youths also responded more similarly than differently, albeit with some variations within specific assessment points in questionnaire data. CONCLUSIONS: Exposure-based CBT for phobic and anxiety disorders produced positive treatment gains and maintenance for Hispanic/Latino youths who participated in the trials. The treatment response that can be expected is generally similar (i.e., favorable) and equivalent to that found with European-American youths based on all the available indices of change. PMID- 14560168 TI - Screen for Child Anxiety Related Emotional Disorders (SCARED): psychometric properties in an African-American parochial high school sample. AB - OBJECTIVE: To examine the psychometric properties of the Screen for Child Anxiety Related Emotional Disorders (SCARED) for a community sample of African-American high school students. METHOD: The 41-item SCARED was administered to 111 adolescents (57 girls; mean age 15.75) in an urban parochial school. Item frequency, internal consistency, test-retest reliability, and construct validity were assessed. RESULTS: Approximately 30% of the sample had scores high enough to warrant further assessment for anxiety disorders, and girls reported significantly higher anxiety symptoms than boys. Internal consistency (alpha =.89) and test-retest reliability (r = 0.47) over 6 months for the SCARED's total score were good. The SCARED's total score was positively correlated with other measures of anxiety symptoms and inattention and was negatively correlated with perceived self-worth. Unlike the five-factor structure reported for primarily white samples, only three factors emerged for this African-American sample. CONCLUSIONS: The SCARED shows utility as a self-report anxiety screening instrument in a community sample of African-American youths. PMID- 14560169 TI - Predictors of urgency in a pediatric psychiatric emergency service. AB - OBJECTIVES: To test the hypothesis that youth present to a psychiatric emergency service (PES) at least 25% of the time for nonurgent reasons, to examine the demographic characteristics that distinguish urgent from nonurgent visits, and to develop a model to predict urgency. METHOD: Psychiatric emergency visits of all patients under 18 years from July 1, 1997, through June 30, 1998, were ed and coded as to level of urgency using Rosenn's classification system. Age, gender, ethnicity, arrival status, social service involvement, violence, substance abuse, and diagnosis were examined with respect to urgency in bivariate and multivariable analyses. RESULTS: Forty percent of visits to the PES were not urgent. Demographic factors distinguished urgent from nonurgent visits. Independent predictors of urgency for the entire sample were age, diagnosis, arrival status, social service involvement, and violence. Only violence remained an independent predictor of urgency for the attention-deficit/hyperactivity disorder subgroup. CONCLUSIONS: A large percentage of pediatric visits to the PES were for nonurgent reasons. This suggests that there are considerable unmet mental health needs of children and adolescents. These findings can be applied to improve and design appropriate services. PMID- 14560170 TI - The British Child and Adolescent Mental Health Survey 1999: the prevalence of DSM IV disorders. AB - OBJECTIVE: To describe the prevalence of DSM-IV disorders and comorbidity in a large population-based sample of British children and adolescents. METHOD: Using a one-phase design, 10,438 children were assessed using the Development and Well Being Assessment (DAWBA), a structured interview with verbatim reports reviewed by clinicians so that information from parents, teachers, and children was combined in a manner that emulated the clinical process. The authors' analysis examined comorbidity and the influence of teacher reports. RESULTS: The overall prevalence of DSM-IV disorders was 9.5% (95% confidence interval 8.8-10.1%), but 2.1% of children were assigned "not otherwise specified" rather than operationalized diagnoses. After adjusting for the presence of a third disorder, there was no longer significant comorbidity between anxiety and conduct disorder or attention-deficit/hyperactivity disorder (ADHD), or between depression and oppositional defiant disorder. A comparison of the disorders in children with and without teacher reports suggested that the prevalence of conduct disorders and ADHD would be underestimated in the absence of teacher information. CONCLUSIONS: Roughly 1 in 10 children have at least one DSM-IV disorder, involving a level of distress or social impairment likely to warrant treatment. Comorbidity reported between some childhood diagnoses may be due to the association of both disorders with a third. Diagnoses of conduct disorder and ADHD may be missed if information is not sought from teachers about children's functioning in school. PMID- 14560171 TI - Genetic and environmental contributions to stability and change in children's internalizing and externalizing problems. AB - OBJECTIVE: To estimate genetic, shared environmental, and nonshared environmental contributions to stability and change in internalizing and externalizing problems. METHOD: Maternal Child Behavior Checklist ratings were obtained for 3,873 twin pairs at age 3 and 1,924 twin pairs at age 7. For 1,575 twin pairs, ratings were available at both ages. RESULTS: For Internalizing/Externalizing ratings, genetic, shared, and nonshared environmental factors explained about 59/51%, 10/30%, and 31/19% of the variance at age 3, and 40/52%, 31/32%, and 29/16% of the variance at age 7. The phenotypic correlation of r = 0.38/0.54 between problems assessed at 3 and 7 years of age was explained for 66/55% by genetic factors, for 23/37% by shared environmental factors, and for 11/8% by nonshared environmental factors. The genetic, shared environmental, and nonshared environmental correlations between ages 3 and 7 were 0.51/0.57, 0.47/0.66, and 0.13/0.24, respectively. CONCLUSIONS: Genetic and shared environmental factors were most important for the stability of Internalizing and Externalizing Problems between ages 3 and 7. Nonshared environmental factors were mainly age-specific. For Internalizing Problems, shared environment may become more important from early to middle childhood. PMID- 14560172 TI - Gastrointestinal distress to serotonergic challenge: a risk marker for emotional disorder? AB - OBJECTIVE: Serotonin is an important mediator of gut sensation and motility. The authors' aim was to determine whether inadvertent gastrointestinal (GI) distress to serotonergic challenge predicted future major depressive and/or anxiety disorders in exposed children. METHOD: l-5-hydroxytryptophan was administered to 119 prepubertal children free of psychiatric disorder as part of a psychobiological cohort study initially designed to examine familial loading for mood disorder as the exposure of interest. Subjects were followed longitudinally with standardized psychiatric interviews to identify new-onset mood and anxiety disorders over 90.3 +/- 29.2 months, with the average assessment interval being 16.6 +/- 6.2 months. Reports of GI distress in a subgroup during serotonergic challenge led the authors to examine GI distress to infusion as an exposure post hoc and to perform survival analysis using major depressive and/or anxiety disorders as the outcomes of interest. RESULTS: GI distress to serotonergic challenge was experienced by 23 subjects, with 7 (30.4%) developing an emotional disorder during follow-up in comparison to 12 (10.4%) of 96 nondistressed subjects. The distressed group was at significantly greater risk of subsequent major depression and/or anxiety (p =.026), even after controlling for family history of psychiatric disorder. CONCLUSIONS: GI distress to serotonergic challenge in childhood is associated with heightened risk for subsequent major depressive and/or anxiety disorders. Studies of serotonergic neurotransmission may aid our understanding of nonrandom associations between functional GI symptoms and emotional symptoms and disorders. PMID- 14560173 TI - Retrospective study of hepatic enzyme elevations in children treated with olanzapine, divalproex, and their combination. AB - OBJECTIVE: To evaluate hepatic enzyme elevations during treatment with olanzapine, divalproex, and their combination. METHOD: Fifty-two children, aged 4 to 18 years, with hepatic enzyme levels measured during treatment with olanzapine (n = 17), divalproex (n = 23), or their combination (n = 12), were identified in the computerized records at a pediatric medical center. Clinical characteristics as well as serial alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase levels were collected. RESULTS: Mean and peak hepatic enzyme levels were significantly higher for the combined treatment group compared to the olanzapine or divalproex groups. All 12 patients who received combined treatment had at least one peak enzyme elevation during the treatment. For 42% of these patients, at least one enzyme level remained elevated during the time for which values were available (mean 8 +/- 6 months). For those treated with divalproex either alone or in combination, the findings were not explained by variations in divalproex plasma levels. Two patients receiving combined treatment had the combination treatment discontinued because of medical complications (pancreatitis in one and steatohepatitis in the other). CONCLUSIONS: Combined treatment with olanzapine and divalproex was associated with more elevations of hepatic enzymes than treatment with either agent alone. The long-term significance of this is unknown but warrants study. PMID- 14560174 TI - A pharmacokinetic/pharmacodynamic study comparing a single morning dose of adderall to twice-daily dosing in children with ADHD. AB - OBJECTIVE: To determine the pharmacokinetic and pharmacodynamic properties of once-daily versus twice-daily doses of Adderall. METHOD: Following a 1-week wash out, 12 subjects with attention-deficit/hyperactivity disorder (ADHD) entered a double-blind crossover study comparing two conditions: QD (10 mg of Adderall at 7:30 a.m. and placebo at noon) or BID (10 mg of Adderall at 7:30 a.m. and at noon). At two sites, cohorts of six subjects each were assessed on two different days by a 12-hour laboratory school protocol. Plasma concentrations of d- and l amphetamine, vital signs, teacher ratings of classroom behavior on the SKAMP, and 10-minute Math Test performance were measured repeatedly over 12 hours. An analysis of variance used center, subject-within-center, condition, and time after-second-dose as independent variables. RESULTS: The pharmacokinetic profiles revealed similar morning concentrations of d- and l-amphetamine. However, concentrations were twice as high in the afternoon for BID as QD. The two conditions showed similar pharmacodynamic profiles in the morning, although improvement in math performance and behavior was maintained into the afternoon only in the BID condition (p <.05). CONCLUSIONS: This study suggests that twice daily dosing of Adderall may be an effective strategy for afternoon control of attention and deportment for children with ADHD. PMID- 14560175 TI - Deficient response inhibition as a cognitive endophenotype of ADHD. AB - OBJECTIVE: To investigate whether a deficient response inhibition is a cognitive endophenotype of attention-deficit/hyperactivity disorder (ADHD). The authors hypothesized that nonaffected siblings of ADHD probands would have a response inhibition between that of ADHD probands and normal controls, although they resembled the controls at a behavioral level. METHOD: Participants were 25 ADHD probands with a family history of ADHD, their nonaffected siblings (n = 25), and 48 normal controls matched for age and IQ. All participants were between 6 and 17 years of age. The nonaffected siblings were compared with their ADHD siblings and with controls on measures reflecting different types of response inhibition. RESULTS: The nonaffected siblings had results similar to those of the ADHD probands, who differed from the controls on all inhibition measures (p <.05). CONCLUSIONS: Siblings of ADHD probands, while not behaviorally expressing the disorder, have ADHD-associated deficits in response inhibition. This suggests that subtyping based on measures of response inhibition can help identify genetic susceptibility to ADHD. Children with a genetic vulnerability to ADHD may have hidden cognitive deficits in the absence of manifest behavioral symptoms. Therefore, they should be monitored to detect possible learning problems. PMID- 14560176 TI - More lessons from a hematology/oncology psychosocial conference: focusing on issues that confront the medical team. PMID- 14560177 TI - Selection and use of inferential statistics: a summary. PMID- 14560178 TI - Genetics of childhood disorders: LV. Prenatal drug exposure. PMID- 14560179 TI - Introduction for scoliosis research society focus issue on motion preservation. PMID- 14560180 TI - Global introduction: SRS focus issue on motion preservation. PMID- 14560181 TI - Adapting innovative motion-preserving technology to spinal surgical practice: what should we expect to happen? AB - STUDY DESIGN: A literature-based review of approach-related morbidity and a conjectural analysis of potential complications of disc arthroplasty based on experience with total joint arthroplasty. OBJECTIVE: To describe predictable complications of disc arthroplasty and possible strategies for minimizing or treating these complications. SUMMARY OF BACKGROUND DATA: There is a significant experience with anterior approach-related morbidity in spinal surgery. There is also extensive experience with extremity total joint arthroplasty. The combination of these experiences should predict certain occurrences that will occur with the advent of disc arthroplasty in the spine. METHODS: Review of the medical literature associated with anterior approach to the lumbar spine for spinal fusion was done. Sequential steps for performance of disc arthroplasty and possible problems with each step were evaluated and possible complications identified. Parallel experience in total joint arthroplasty was reviewed for possible predictive experience. RESULTS: There are definable approach-related morbidities that will occur, regardless of prosthesis design and implantation technique. Prosthesis design involves a series of tradeoffs for risks and benefits. Revisions are inevitable; rate of revision and time to revision remain to be determined. CONCLUSIONS: Disc arthroplasty will offer benefits over current fusion techniques. It will come at a cost and certain complications are entirely predictable. There will be deaths from the procedure, due to thromboembolic phenomenon or due to uncontrollable hemorrhage from irreparable vascular injury, especially on repeat operations. There will be prostheses that dislodge. There will be infections that require device removal, a very high-risk procedure. There will be a deterioration of results in the hands of the general medical community as opposed to the hands of the initial investigators, a learning curve if you will. The access surgeon will be critical to minimizing morbidity. Design considerations compete with anatomic constraints. Material choices all have pros and cons. Spine surgeons as a whole are excited about this opportunity, but we must be diligent to minimize these predictable adverse events to make the risk benefit profile the best that it can be for our patients. PMID- 14560182 TI - Biomechanical evaluation of total disc replacement arthroplasty: an in vitro human cadaveric model. AB - STUDY DESIGN: This in vitro biomechanical study was undertaken to quantify the multidirectional intervertebral kinematics following total disc replacement arthroplasty compared to conventional stabilization techniques. OBJECTIVE: Using an in vitro human cadaveric model, the primary objective was to compare the multidirectional flexibility properties and map the center of intervertebral rotation of total disc arthroplasty versus conventional threaded fusion cages and cages augmented with transpedicular fixation for single-level spinal instrumentation. SUMMARY OF BACKGROUND DATA: The utilization of motion-preserving implants versus instrumentation systems, which stabilize the operative segments, necessitates improved understanding of their comparative biomechanical properties. METHODS: A total of eight human cadaveric lumbosacral spines (L2 to sacrum) were utilized in this investigation and biomechanically evaluated under the following L4-L5 reconstruction conditions: 1) intact spine; 2) SB Charite disc prosthesis; 3) BAK cages; and 4) BAK cages + ISOLA pedicle screw/rod fixation (anteroposterior). The superior (L3-L4) and inferior (L5-S1) intervertebral levels remained uninstrumented to quantify adjacent level properties. Multidirectional flexibility included pure, unconstrained moments (+/ 8 Nm) in axial rotation, flexion-extension, and lateral bending, with quantification of the operative and adjacent level range of motion and neutral zone, which were normalized to the intact spine condition. RESULTS: The SB Charite prosthesis indicated an average percentage increase in axial rotation range of motion by 44% compared to the intact condition (P < 0.05), whereas the BAK and anteroposterior reconstructions decreased range of motion by 29% and 80%, respectively (P < 0.05). The SB Charite was significantly different from BAK and combined anteroposterior reconstructions (P < 0.05). Flexion-extension indicated a minor increase in range of motion for the SB Charite (3%) versus the intact disc (P > 0.05), whereas the BAK and anteroposterior stabilization groups resulted in significant decreases in range of motion (BAK = 57%, anteroposterior = 93%) (P < 0.05) when compared to the intact and SB Charite conditions. Based on flexion-extension radiographs, the intervertebral centers of rotation were in the posterior one-third of the operative intervertebral disc only for the SB Charite reconstruction and intact spine condition, with definitive evidence of physiologic intervertebral translation (intact 2.06 +/- 77 mm; SB III = 1.9 +/- 0.98 mm). CONCLUSIONS: Total disc arthroplasty serves as the next frontier in the surgical management of discogenic spinal pathology. The SB Charite restored motion to the level of the intact segment in flexion-extension and lateral bending and increased motion in axial rotation. The anterior annular resection necessary for device implantation and unconstrained design of the prosthesis account for this change in rotation. The normal lumbar flexion-extension axis of rotation is an ellipse rather than a single point. Only disc replacement rather than pedicle instrumentation or BAK interbody instrumentation preserves the kinematic properties and normal mapping of segmental motion at the operative and adjacent intervertebral disc levels. PMID- 14560183 TI - General principles of total disc replacement arthroplasty: seventeen cases in a nonhuman primate model. AB - STUDY DESIGN: To investigate the biomechanical, histochemical, and biologic ingrowth characteristics of two different lumbar disc prostheses-AcroFlex (DePuy AcroMed) and the SB Charite (DePuy-AcroMed)-for total disc replacement arthroplasty. METHODS: A total of 17 mature baboons (n = 17, Papio cynocephalus) underwent L5-L6 total disc replacement procedures. The AcroFlex device (n = 10 levels) consisted of sintered titanium beaded ingrowth surfaces, bound together by a hexene-based polyolefin rubber core. The SB Charite (n = 7 levels) device prosthetic vertebral end plates were cobalt chrome, covered by two layers of thin titanium with an electrochemically bonded hydroxyapatite coating and an ultra high molecular weight polyethylene core. RESULTS: Following 6-month survival periods, the range of motion of the SB Charite and intact nonoperative controls under axial compression, flexion-extension, and lateral bending showed no statistical difference (P > 0.05). However, both exhibited greater range of motion compared to the AcroFlex treatments (P < 0.05). Plain film radiographic analysis showed no lucencies or loosening of any metallic prosthetic vertebral endplate. Gross histopathologic analysis of the AcroFlex and SB Charite prosthesis demonstrated excellent ingrowth at the level of the implant-bone interface, without evidence of fibrous tissue or synovium. Histochemical assays showed no local or systemic accumulation of particulate wear debris (titanium, ultra-high molecular weight polyethylene, or cobalt chrome) nor cytokines (TNF alpha, PGE2, IL-1, IL-2, or IL-6). Porous ingrowth calculations showed the mean ingrowth (linear apposition) ranging from 47.9% +/- 9.12 for the SB Charite device and 54.59% +/- 13.24 for the AcroFlex device. CONCLUSIONS: The porous ingrowth, percentage pore ingrowth coverage at the bone-metal interface was more favorable for total disc replacement compared to that reported for cementless total joint components in the appendicular skeleton (range 10-30%). The reason for the improved degree of porous ingrowth in total disc replacement prostheses is probably due to ligamentotaxis causing sustained compression across the metal bone interface. This project serves as the first comprehensive in vivo investigation comparing two different types of unconstrained disc prostheses with alternate in-growth surfaces and establishes an excellent research model in the evaluation of lumbar total disc replacement arthroplasty. PMID- 14560184 TI - Spinal implant debris-induced osteolysis. AB - STUDY DESIGN: Generally, implant-induced osteolysis is a manifestation of an adverse cellular response to phagocytosable particulate wear and corrosion debris. Initially termed "cement disease," particle-induced loosening was recognized by Charnley in the early 1960s. Despite the plethora of information gained over the last 40 years on the basic science of periprosthetic bone loss, much remains unanswered. The effect of unintended debris resulting from wear and corrosion (e.g., micromotion between the interconnection mechanisms in spinal implants) remains a clinical concern. The current study highlights what is known of particle-induced osteolysis and how the presence of spinal implant particulate debris deleteriously influences osseointegration of posterolateral bone graft or disrupts an established posterolateral fusion mass. Tissue explant, animal, and cell culture studies have revealed the complexity of cellular reactivity involved in aseptic particle-induced osteolysis. OBJECTIVES: The objectives of this study are twofold: 1) to highlight the dominant cellular participants in total joint arthroplasty particle induced osteolysis, which are purportedly the macrophage, osteoblast, fibroblast, and osteoclast and several of the dominant chemical mediators have been identified as well, which include prostaglandin E2, tumor necrosis factor-alpha, interleukin-1, and interleukin-6; and 2) to demonstrate the potential deleterious effects of spinal implant debris using animal models and analysis of soft tissue surrounding spinal implants in symptomatic patients. METHODS: There are a growing number of proinflammatory and anti-inflammatory cytokines, prostenoids, and enzymes that have been shown to play important roles in the pathology of particle-induced osteolysis. Reports that aseptic granulomatous inflammation typical of that associated with corrosion debris appear to correlate with the complexity of the implant. Titanium particulate material was used to induce effects in 34 New Zealand White rabbits where analysis included serological quantification of systemic cytokines. Postmortem microradiographic, immunocytochemical, and histopathologic assessment of the intertransverse fusion mass quantified the extent of osteolysis, local proinflammatory cytokines, osteoclasts and inflammatory infiltrates. Clinical analysis of 12 patients more than 0.4 years after spinal implants (mean 4.03, range 0.4 to 11 years) presented with late operative site pain. RESULTS: Currently the etiology of this inflammation around spinal implants resembles particle-induced osteolysis around joint arthroplasties where there typically is a self-perpetuating fibroinflammatory zone adjacent to the implant, where macrophage exhaustion, reactive oxygen intermediates, and pro-inflammatory cytokines affect a host of local cell types and induce a widening zone of soft tissue damage and inflammation. Animal model analysis indicated increased levels of local inflammatory cytokines typically associated with osteolysis-tumor necrosis factor-alpha. Osteoclast cell counts and regions of osteolytic resorption lacunas were higher in the titanium-treated versus autograft-alone groups (P < 0.05), and the extent of cellular apoptosis was markedly higher in the titanium-treated sites at both time intervals. Electron microscopy indicated definitive evidence of phagocytized titanium particles and foci of local, chronic inflammatory changes in the titanium-treated sites. CLINICAL CASES: 11 of 12 clinical cases demonstrated elevated tumor necrosis factor-alpha levels and an increased osteoclastic response in the vicinity of wear debris caused by dry frictional wear particles of titanium or stainless steel. Resection of the wear debris and surrounding fibroinflammatory zone resolved clinical symptoms in all 12 cases. CONCLUSIONS: More basic science and clinical research is needed to develop novel strategies for gaining knowledge, and developing effective evaluation and treatment of patients with implant debris related osteolysis. Titanium debris simulating that produced by spinal implants introduced at the level of a spinal arthrodesis elicits an inflammatory cytokine mediated particulate-induced response through increased expression of intracellular TNF alpha, increased osteoclastic activity and cellular apoptosis. This study highlighted the association between spinal implants particulate wear debris and increased potential for osteolysis. Aseptic osteolysis is among the primary reasons for failure of orthopedic implants. Increased awareness of this destructive process is becoming more important with the growing popularity of total disc arthroplasty and highly modular spinal implants. PMID- 14560185 TI - Biomaterial optimization in total disc arthroplasty. AB - STUDY: Knowledge gained through the clinical history of total joint replacement materials combined with the current promise of new biomaterials provides improved guidelines for biomaterial selection in total disc arthroplasty. OBJECTIVES: The following will detail: 1) current biomaterials technology; 2) how current designs of total disc arthroplasty seek to optimize implant performance through judicious biomaterial selection; and 3) what technical obstacles and clinical concerns remain. METHODS: Metals and polymers remain the central material components of state-of-the-art total joint arthroplasties. Polymers provide low friction surfaces for articulating bearings and some degree of shock absorption. Metals provide appropriate material properties such as high strength, ductility, fracture toughness, hardness, corrosion resistance, formability, and biocompatibility necessary for use in load-bearing roles required total disc replacement. There are three principal metal alloys used in orthopaedics and particularly in total joint replacement: 1) titanium based alloys; 2) cobalt based alloys; and 3) stainless steel alloys. Alloy specific differences in strength, ductility, and hardness generally determine which of these three alloys is used for a particular application or implant component. RESULTS: Current designs. Two examples of current lumbar (Charite and Prodisc) and cervical (Bryan and Prestige) disc replacements are compared. The similarities and differences in the biomaterials used for each demonstrate prevailing consensus and some idea of how to best optimize implant performance through biomaterial selection. CONCLUSION: The primary factors governing total disc arthroplasty biomaterials are similar to those of all total joint arthroplasties: generation of wear debris is the primary source of implant degradation, and the subsequent tissue reaction to such debris is the primary factor limiting the longevity of joint replacement prostheses. Particulate debris generated by wear, fretting, or fragmentation induces the formation of an inflammatory reaction, which at a certain point promotes a foreign-body granulation tissue response that has the ability to invade the bone-implant interface. This commonly results in progressive, local bone loss that threatens the fixation of both cemented and cementless devices alike. All metal alloy implants corrode in vivo. When severe, the degradative process may reduce structural integrity of the implant, and the release of corrosion products is potentially toxic to the host. The corrosion resistance of implant alloys is primarily due to the formation of passive oxide films to prevent significant electrochemical dissolution from taking place. The result of this knowledge is a consensus of opinion as to which materials are best suited for use in current total disc arthroplasty designs, where most total disc replacement designs incorporate cobalt-chromium-molybdenum alloy endplates articulating internally on a relatively soft polymeric core and externally coated with titanium or titanium alloy for enhanced bone fixation. PMID- 14560186 TI - Experimental design of total disk replacement-experience with a prospective randomized study of the SB Charite. AB - STUDY DESIGN: Prospective randomized clinical trial. OBJECTIVES: To determine if a prospective randomized study of patients with symptomatic degenerative disc disease treated with disc arthroplasty could be safely completed. METHODS: Sixty patients with one-level discogenic pain confirmed by plain radiography, magnetic resonance imaging, and provocative discography for degenerative disc disease were randomized comparing 1/3 BAK anterior interbody fusion and 2/3 anterior SB Charite artificial disc replacement. RESULTS: The mean age was 40.3 years (range 21-56 years). Nineteen cases were at L4-L5 and 41 cases were at L5-S1. Nineteen cases had BAK anterior interbody fusion and 41 cases were randomized as SB Charite disc replacement. The length of surgery was mean 88.4 minutes (range 54 137 minutes) for both groups. The estimated blood loss was mean 289.5 cc (range 50-1800 cc). The length of hospital stay was a mean of 3.03 days (range 2 to 6 days). Oswestry Disability Index for the BAK control group was 45.9 +/- 10.4 before surgery and 23.5 +/- 17.2 at follow-up (P < 0.001). The corresponding ODI scores for the SB Charite disc were 50.0 +/- 14.3 before surgery and 25.0 +/- 20.1 at a mean of 2 years' follow-up (P < 0.001). CONCLUSIONS: This is the first study that shows improvement of functional outcome measures in a prospective randomized design treating primarily mechanical back pain and achieving comparable successful results to lumbar spinal stenosis decompression. PMID- 14560187 TI - Clinical results with ProDisc: European experience and U.S. investigation device exemption study. AB - STUDY DESIGN: This study is based on a review of the literature related to the use of the ProDisc device and a report of the preliminary results of a prospective randomized study. OBJECTIVES: To review European results related to the use of the ProDisc device and compare the results of this device to lumbar fusion in a prospective, randomized study being performed as part of a Food and Drug Administration-approved investigational study. SUMMARY OF BACKGROUND DATA: There are two devices that have been used on a large-scale basis for total disc replacement. These are the SB Charite and the ProDisc. Both devices were created in Europe and have been used there for more than 10 years. Reported results for these devices have been favorable, but there have been no prospective studies evaluating the outcome. METHODS: The literature related to the ProDisc was reviewed. The preliminary study data were based on one center's experience participating in the Food and Drug Administration Investigation Device Exemption study. There were 39 patients with a minimum 6-month follow-up. Patients were randomly assigned to receive either the ProDisc or undergo a combined anterior posterior lumbar fusion in a ratio of 2:1 (ProDisc to fusion). Patients completed standardized questionnaires before surgery and at 6 weeks, 3 months, and 6 months after surgery. Data collection is continuing for the 12- and 24-month follow-up. RESULTS Operative time, blood loss, and length of hospitalization were significantly less in the disc replacement group (P < 0.05). At the 3-month follow-up, the disc replacement group had a significantly greater improvement in Oswestry scores than did the fusion group. There were no differences in pain scores as measured by visual analog scales. Disc replacement patients had greater motion and there was a trend for this group to have greater satisfaction at the 6 month follow-up. CONCLUSIONS: The preliminary results of this prospective randomized study found that peri-operative factors were more favorable in the disc replacement group than in the fusion group. There was a trend to greater patient satisfaction in this group. These early results suggest that total disc replacement may be a viable alternative to lumbar spinal fusion in patients with symptomatic disc disruption unresponsive to nonoperative care. Long-term follow up is needed and is currently being collected for this study group. PMID- 14560188 TI - ProDisc artificial total lumbar disc replacement: introduction and early results from the United States clinical trial. AB - STUDY DESIGN: Multicenter prospective randomized study of artificial disc replacement (ProDisc) versus circumferential fusion (standard of care) for one- and two-level degenerative disc disease. This is an interim analysis on patients seen at the Spine Institute Saint John's Health Center, Santa Monica, California. OBJECTIVES: To evaluate early pain and functional outcomes of patients treated with disc replacement or fusion and to assess the capacity of this intervertebral disc replacement for preserving motion in the lumbar spine. SUMMARY OF BACKGROUND DATA: Disc replacement is intended to reduce pain via removal of the diseased disc while restoring physiologic motion and height at the affected level. The long-term physiologic advantage of disc replacement to fusion is that preservation of motion may prevent additional degeneration at adjacent levels. METHODS: Patients meeting inclusion criteria were consented for study. Randomization was performed using a 2 to 1 ratio of disc replacement procedure to a fusion procedure. Patients rated their pain on the Visual Analogue Scale and completed the Oswestry Disability Index questionnaire. Radiographs were taken. Assessments were made before surgery and after surgery at 6 weeks, 3 months, 6 months, and 1 year (ongoing). Changes from preoperative pain, disability, or motion were separately evaluated as a function of treatment using repeated measures mixed design analysis of variance. RESULTS: This analysis includes data up to 6 months from the first 53 randomized patients. There were 35 patients who underwent disc replacements, and 18 patients had fusion procedures. Disc replacement patients had a significant reduction in pain and disability at earlier evaluations. By 6 months, the relative improvement on both the Visual Analogue Scale and Oswestry (both, P < 0.05) were similar for disc replacement and fusion patients. Greater motion was found at L4-L5 for disc replacement patients (P < 0.05) than fusion patients. A similar trend was noted at L5-S1 (P was not significant). CONCLUSIONS: Disc replacement patients reported significantly less pain (Visual Analogue Scale) and disability (Oswestry) in the early period following surgery compared to fusion patients. This difference disappeared by 6 months. When compared to fusion, the disc replacement allowed preservation of motion at L4-L5 with a similar trend at L5-S1. PMID- 14560189 TI - Cervical disc replacement-porous coated motion prosthesis: a comparative biomechanical analysis showing the key role of the posterior longitudinal ligament. AB - STUDY DESIGN: Benchtop cadaveric biomechanical comparative testing and caprine animal model in vivo implantation. OBJECTIVE: To evaluate the role of the posterior longitudinal ligament in cervical arthroplasty and to understand the relative contribution of this ligament in nonfusion applications. SUMMARY OF BACKGROUND DATA: Rauschning refers to the posterior longitudinal ligament as "The Kleenex Ligament" due to its apparent anatomic insignificance. White and Panjabi found the posterior longitudinal ligament ranked only fourth in importance in tensile load-to-failure biomechanical testing. In the postoperative situation following anterior cervical diskectomy fusion, posterior longitudinal ligament integrity is overlooked by physicians because the entire disc space usually fuses into a homogeneous block of bone. PURPOSE: This biomechanical study was undertaken to determine the relative importance of the posterior longitudinal ligament following two different degrees of anterior decompression, anterior disc replacement, and anterior arthrodesis procedures. METHODS: A total of seven fresh frozen human cadaveric cervical spines (C3-C7) (mean age 68 +/- 19 years) were used for biomechanical testing. Each vertebra was equipped with three non colinear light emitting diodes designed for detection by an optoelectronic motion measurement system (3020 Optotract System). To determine the multidirectional flexibility, six pure moments (flexion, extension, right + left lateral bending, right + left axial rotation) and axial compression were applied using a servohydraulic 858 Bionix testing device configured with a six-degree-of-freedom spine simulator. Range of motion was defined as the peak displacement from the initial neutral position to the maximum load, whereas the neutral zone represents the motion from the initial neutral position to the unloaded position at the beginning of the third cycle. Seven groups of (N = 7 each) constructs at C5-C6 were: 1) intact "native" C5-C6 level; 2) anterior diskectomy (posterior longitudinal ligament intact); 3) a Low Profile Porous Coated Motion cervical disc replacement; 4) posterior longitudinal ligament resected; 5) Porous Coated Motion cervical disc replacement fixed with anterior flanges and screws; 6) tricortical structural allograft; and 7) an anterior cervical translational plate + allograft. The caprine model was evaluated for suitability as an animal model with 12 goats undergoing C3-C4 anterior cervical Porous Coated Motion disc replacement. RESULTS: Group 2 (anterior diskectomy alone) was significantly more stable than Group 4 (anterior diskectomy + posterior longitudinal ligament resection) in flexion-extension, 18.7 +/- 4.76 degrees versus 24.8 +/- 4.42 degrees (P < 0.05) and in lateral bending, 5.9 +/- 1.79 degrees versus 10.7 +/- 2.8 degrees (P < 0.05). The comparison for the two conditions for axial rotation, 10.4 +/- 13.9 degrees versus 13.9 +/- 2.7 degrees, and axial compression, 1.19 +/ .98 degrees versus 1.52 +/- 1.14 degrees, showed the same trend. Twelve goats undergoing porous coated motion cervical disc replacement had no evidence of prosthesis loosening, neurologic complications, or experienced inflammatory reactions from particulate wear debris after 6 months of implantation. DISCUSSION: This study confirms the pivotal role of the posterior longitudinal ligament in postsurgical stability of the cervical spine following anterior diskectomy. This is because the lateral anulus, uncovertebral ligaments, and lateral capsular ligaments are stretched and plastically deformed in the surgical distraction process of restoring the disc space height following anterior surgical decompression. There should be a separate determination of the range of motion of cervical disc replacements depending of the integrity and the amount of the posterior longitudinal ligament that has been resected. CLINICAL RELEVANCE: There are two basic types of total knee replacements, posterior cruciate ligament preserving and posterior cruciate ligament-sacrificing designs. In the cervical spine, an analogous situation exists biomechanically depending on whether the posterior longitudinal ligament needs to be removed in its entirety as part of the spinal cord decompression part of the procedure--it may be helpful to conceptually differentiate between posterior longitudinal ligament-preserving and posterior longitudinal ligament-sacrificing total cervical disc replacements. PMID- 14560191 TI - Basic science summary statement. PMID- 14560190 TI - Wear analysis of the Bryan Cervical Disc prosthesis. AB - STUDY DESIGN: In vitro wear testing of the Bryan Cervical Disc prosthesis was performed in a cervical spine simulator. The biologic response was assessed in chimpanzee and goat animal models. OBJECTIVE: Determine the wear characteristics of the Bryan disc. SUMMARY OF BACKGROUND DATA: Large joint arthroplasties fail most commonly by wear and consequent formation of particulate material, which induces an inflammatory response. Therefore, measuring the wear characteristics of the new spinal disc replacements is important. METHODS: Six prosthetic assembles were tested to 10 or 40 million cycles by load and motion and 3 additional assemblies were tested by load only in a cervical spine simulator. Any debris was examined using ASTM standards. The local biologic response to the prosthesis was examined in two chimpanzees. Nine goats were used to assess the biologic response in both local and distant tissues. Arthrodesis was performed on three additional control goats that received an allograft and an anterior cervical plate. RESULTS: Wear results: cervical spine simulators that applied the loads and motions associated with activities of daily living produced wear particulate at a rate of 1.2 mg per million cycles. Device height decreased 0.02 mm per million cycles with approximately 77% of this decrease due to gradual creep of the nucleus under the constant compressive load. Particles generated were granular in shape with a mean feret diameter of 3.9 microm. All animals tolerated placement of the Bryan disc. Wear debris was present in the periprosthetic and epidural spaces in some animals. However, no significant inflammatory response was observed. No wear material was found distant from the implant in draining lymph tissue, the liver, or the spleen. CONCLUSIONS: The Bryan disc has satisfactory wear characteristics and does not produce a significant inflammatory response. PMID- 14560192 TI - Clinical summary statement. PMID- 14560193 TI - The Lenke classification of adolescent idiopathic scoliosis: how it organizes curve patterns as a template to perform selective fusions of the spine. AB - STUDY DESIGN: Retrospective radiographic review. OBJECTIVES: To analyze how the Lenke classification of adolescent idiopathic scoliosis provides a template of specific curve patterns that may be appropriate to perform selective fusion of the spine. METHODS: A new triad classification system of adolescent idiopathic scoliosis has been developed. It consists of a curve type, a lumbar spine modifier (A, B, C), and a sagittal thoracic modifier (-, N, +). A selective fusion is termed when both the thoracic and thoracolumbar/lumbar curves deviate completely from the midline, but only the major curve (largest Cobb measurement) is fused, leaving the minor curve unfused and mobile. In this manner, selective thoracic fusions of the spine are potentially indicated for major main thoracic/minor lumbar curves (Types 1C and potentially 2C and 3C patterns) when the lumbar apex deviates off the center sacral vertical line. Conversely, selective thoracolumbar/lumbar fusions may be indicated for major thoracolumbar/lumbar-minor main thoracic curves, when the thoracic apex lies off the C7 plumbline (Type 5C and potentially 6C patterns). Importantly, additional analysis of ratios of structural characteristics between the main thoracic and thoracolumbar/lumbar curves are necessary to predict when a successful selective main thoracic or thoracolumbar/lumbar fusion will be feasible. Lastly, the clinical appearance of the patient's truncal alignment is essential to confirm the aspirations of performing a selective spinal fusion. RESULTS: Successful selective thoracic fusion of 1C (n = 36) and 2C (n = 8) curves have been performed in 44 consecutive patients with adolescent idiopathic scoliosis. The average thoracic curve was 61 degrees before surgery and 39 degrees at final follow-up. The average preoperative lumbar curve was 48 degrees, decreasing to 32 degrees postoperatively. A group of 21 consecutive patients with Type 5C or 6C major thoracolumbar/lumbar-minor main thoracic curves underwent a selective thoracolumbar/lumbar fusion. The average preoperative thoracolumbar/lumbar curve was 56 degrees corrected to 22 degrees at the 2-year follow-up. The average minor main thoracic curve preoperative was 38 degrees, with spontaneous correction to 28 degrees at 2 years postoperative. DISCUSSION: Selective thoracic or thoracolumbar/lumbar fusion can be successfully performed in a variety of adolescent idiopathic scoliosis curve patterns. Careful attention to the preoperative Lenke curve classification, analysis of structural characteristics between the planned instrumented and noninstrumented regions of the spine, as well as a documented clinical examination that confirms the planned instrumented and fused regions of the spine to be the most clinically prominent are essential features to determine before surgery. No patients undergoing selective thoracic fusion have required extension of the fusion to the lumbar spine, whereas one patient with a selective thoracolumbar fusion required extension of the fusion up to include the thoracic spine due to continued thoracic progression with growth. CONCLUSIONS: Selective thoracic or thoracolumbar/lumbar fusions of the major curve can be successfully performed even when the minor curve completely deviates from the midline, based on the Lenke classification system, the analysis of structural criteria between the planned fused and unfused regions of the spine, and the clinical examination of the patient. Selective fusions, when successfully performed, will optimize mobile segments of the spine in patients with adolescent idiopathic scoliosis. PMID- 14560194 TI - Anterior single-rod instrumentation of the thoracic and lumbar spine: saving levels. AB - STUDY DESIGN: To evaluate the ability of single-rod anterior instrumentation to save or preserve fusion levels and improve thoracic hypokyphosis in patients with adolescent idiopathic thoracic, thoracolumbar, or lumbar scoliosis. OBJECTIVE: To provide indications for single anterior rod instrumentation for the treatment of adolescent idiopathic scoliosis and demonstrate effectiveness in properly selected cases. SUMMARY OF BACKGROUND DATA: Posterior multisegmented dual rod instrumentation is the most commonly used instrumentation for the surgical treatment of adolescent idiopathic scoliosis. The issue of longer fusion levels and inability to correct hypokyphosis with posterior instrumentation continues to be debated in the literature. Anterior instrumentation has the ability in certain curve patterns to preserve distal and proximal levels as well as correct thoracic hypokyphosis. METHODS: A brief discussion of the Lenke adolescent idiopathic scoliosis classification system is presented. Surgical treatment options for each of the curve types are discussed in detail. RESULTS: Single-rod anterior instrumentation for adolescent idiopathic scoliosis will predictably save levels in Type I curves without hyperkyphosis as well as Type 5 curves; however, it is usually contraindicated in Type 2, Type 4, and Type 6 curves. Single-rod anterior instrumentation can occasionally be utilized in Type 3 curves if the magnitude of the lumbar curve is significantly less than the thoracic curve and the flexibility of the lumbar curve approaches 25 degrees on the side-bending radiograph. CONCLUSIONS: Single-rod anterior instrumentation will often saved one to three distal fusion levels when treating isolated major thoracic, thoracolumbar, or lumbar curves. Fusion levels should include upper to lower Cobb levels. Additionally, anterior single-rod instrumentation because its kyphogenic nature will predictably correct hypokyphosis of the thoracic spine. PMID- 14560196 TI - Short segment bone-on-bone instrumentation for single curve idiopathic scoliosis. AB - STUDY DESIGN: Retrospective case series review. OBJECTIVES: To evaluate the outcomes of a new short segment anterior scoliosis technique with complete removal of the discs, bone-on-bone apposition of the vertebral bodies, and dual rod instrumentation. To evaluate a new preop planning technique for scoliosis instrumentation. SUMMARY OF BACKGROUND DATA: Scoliosis surgery traditionally was performed via a posterior approach, but anterior scoliosis instrumentation has proven to be superior regarding the amount of curve correction and the number of segments saved from instrumentation. METHODS: Thirty-one patients with single curve idiopathic scoliosis less than 75 degrees were operated using the bone-on bone surgical technique with dual rod instrumentation (Kaneda Anterior Scoliosis System, Depuy AcroMed, Raynham, MA from 1996 until 2001). Average follow-up was 40 months (range 15-77 months). RESULTS: Surgical correction of the major curve averaged 73.9% over the instrumented levels and 51.4% over the entire curve. The average number of discs fused was 4.6 for thoracic curves and 3.3 for thoracolumbar and lumbar curves. There were no implant-related complications or nonunions. The compensatory curves spontaneously improved by an average of 38.6%. Uneventful healing of all fusions occurred-most within 8 to 12 weeks. One compensatory thoracic curve progressed and posterior instrumentation was done 28 months after correction of the major thoracolumbar curve. CONCLUSIONS: Surgical correction was achieved in over half the levels that would have been operated by standard posterior segmental fixation. Bony healing due to the bone-on-bone apposition was achieved uneventfully after apical correction of the spinal curvature in all patients. Use of dual rod instrumentation (Kaneda Anterior Scoliosis System) is fundamental in maintaining the correction of the curvature achieved in the operating room. The preoperative planning technique worked well. PMID- 14560195 TI - Factors involved in the decision to perform a selective versus nonselective fusion of Lenke 1B and 1C (King-Moe II) curves in adolescent idiopathic scoliosis. AB - STUDY DESIGN: A retrospective evaluation of 203 adolescent idiopathic scoliosis patients with Lenke 1B or 1C (King-Moe II) type curves. OBJECTIVES: To evaluate the incidence of inclusion of the lumbar curve in the treatment of this type of deformity as well as radiographic factors associated with lumbar curve fusion. SUMMARY OF BACKGROUND DATA: In patients with structural thoracic curves and compensatory lumbar curves, many authors have recommended fusing only the thoracic curve (selective thoracic fusion). Studies have shown that correction of the thoracic curve results in spontaneous correction of the unfused lumbar curve; however, in some cases, truncal decompensation develops. Though there have been various attempts to define more accurately what type of curve pattern should undergo selective fusion, controversy continues in this area. METHODS: Measurements were obtained from the preoperative standing posteroanterior and side-bending radiographs of 203 patients with Lenke Type 1B or 1C curves from five sites of the DePuy AcroMed Harms Study Group. Patients were divided into two groups depending on their most distal vertebra instrumented: the "selective thoracic fusion" group included patients who were fused to L1 or above and the "nonselective fusion" group included patients fused to L2 or below. A statistical comparison was conducted to identify variables associated with the choice for a nonselective fusion. RESULTS: The incidence of fusion of the lumbar curve ranged from 6% to 33% at the different patient care sites. Factors associated with nonselective fusion included larger preoperative lumbar curve magnitude (42 +/- 10 degrees vs. 37 +/- 7 degrees, P < 0.01), greater displacement of the lumbar apical vertebra from the central sacral vertical line, (3.1 +/- 1.4 cm vs. 2.2 +/ 0.8 cm, P < 0.01), and a smaller thoracic to lumbar curve magnitude ratio (1.31 +/- 0.29 vs. 1.44 +/- 0.30, P = 0.01). CONCLUSIONS: The characteristics of the compensatory "nonstructural" lumbar curve played a significant role in the surgical decision-making process and varied substantially among members of the study group. Side-bending correction of the lumbar curve to <25 degrees (defining these as Lenke 1, nonstructural lumbar curves) was not sufficientcriteria to perform a selective fusion in some of these cases. The substantial variation in the frequency of fusing the lumbar curve (6% to 33%) confirms that controversy remains about when surgeons feel the lumbar curve can be spared in Lenke 1B and 1C curves. Site-specific analysis revealed that the radiographic features significantly associated with a selective fusion varied according to the site at which the patient was treated. The rate of selective fusion was 92% for the 1B type curves compared to 68% for the 1C curves. PMID- 14560197 TI - The pros and cons to saving the L5-S1 motion segment in a long scoliosis fusion construct. AB - STUDY DESIGN: This is a review of the literature and personal experience as it pertains to whether a long fusion should be stopped at L5 or S1 in a patient with adult lumbar scoliosis and degenerative changes. OBJECTIVES: To summarize the problems with decision-making and to point out the strengths and limitations of past studies. SUMMARY OF BACKGROUND DATA: There is a paucity of data on this subject. Problems with stopping at L5 include fixation at that segment and subsequent breakdown at L5-S1. The problems with stopping at the sacrum include the additional surgical requirements and increased potential for pseudarthrosis. METHODS: Summarized is past literature and, to some extent, personal experience of the author(s). RESULTS: There are situations where it is clearly preferable to stop at the sacrum. However, there are many borderline circumstances in which whether it is better to stop at L5 or the sacrum is not clear cut. CONCLUSIONS: The answer to this question requires further study. Multicenter data collection, consistency of approach, and potential randomization in a prospective fashion might help provide an answer. PMID- 14560198 TI - Summary statement: fusion technologies. PMID- 14560199 TI - Selective anterior fusion and instrumentation for the treatment of neuromuscular scoliosis. AB - STUDY DESIGN: A retrospective cohort study was conducted. OBJECTIVE: To evaluate the results of anterior spinal fusion with anterior instrumentation alone in selected patients with neuromuscular scoliosis. SUMMARY OF BACKGROUND DATA: Traditionally posterior spinal fusion with instrumentation has been done, usually to the pelvis, to achieve correction of neuromuscular scoliosis. However, certain selected patients might benefit from shorter fusion segment to preserve some motion and yet still achieve good correction of the curve. This may serve to improve or preserve various functional abilities that might be adversely affected by a long fusion. METHOD: Patients who had anterior spinal fusion (ASF) with anterior instrumentation (AI) alone were selected from an entire group of patients with neuromuscular spinal deformity who had surgery at Shriners Hospital for Children-Chicago since January of 1988. The charts and radiographs of these patients were examined and various radiographic parameters were measured pre- and after surgery and at final follow-up. Additionally, functional level of the patients included, ambulatory status was obtained from the medical records. RESULTS: In these 21 patients excellent results were obtained with regard to primary and secondary curve correction as well as the pelvic obliquity without significant deterioration at final follow-up. Ambulatory status was not changed after surgery. This cohort of patients had various neuromuscular diseases. However, the majority of them had myelomeningocele. Few complications occurred which resulted in the reoperation of several patients who had progression of the curve around the instrumented segment which itself remained unchanged when the complication was recognized. One infection occurred requiring irrigation and debridement. CONCLUSIONS: In selected patients with neuromuscular scoliosis, even that associated with pelvic obliquity, excellent correction and maintenance correction can be obtained fusing a relatively short segment of the spine with ASF and AI rather than a long construct posteriorly to the pelvis. Maintenance of the correction of the primary curve as well as the pelvic obliquity was maintained over the period of follow-up. This approach for selected patients should be offered as a way of limiting the extend of the surgery, preserving motion segments and maintaining orenhancing functions such as activities of daily living. PMID- 14560200 TI - Use of video-assisted thoracoscopic surgery to reduce perioperative morbidity in scoliosis surgery. AB - STUDY DESIGN: A case series of idiopathic scoliosis patients treated with thoracoscopic anterior instrumentation was compared to a similar group of patients treated by open anterior instrumentation. OBJECTIVES: To evaluate the morbidity associated with thoracoscopic instrumentation compared to the open approach for thoracic scoliosis. METHODS: A consecutive group of thoracoscopically treated patients with Lenke 1 adolescent idiopathic scoliosis was compared to similar patients gathered from the DePuy-AcroMed Harms Study Group database. Perioperative outcome measures as well as early postoperative functional outcomes (pulmonary function, shoulder strength) were compared. RESULTS: There were 38 thoracoscopic instrumentation cases with greater than 6 months' follow-up that were compared to 68 anterior open instrumentation cases. The radiographic outcomes were similar (60% +/- 11% vs. 59% +/- 17% thoracic curve correction for the thoracoscopic and open groups, respectively). The reduction in forced vital capacity was significantly (P = 0.01) greater in the open group (0.6 +/- 0.3 L) compared to the endoscopic group (0.4 +/- 0.3 L). There was a trend towards greater return of shoulder girdle strength and range of motion 6 weeks after surgery in the thoracoscopic patients. CONCLUSION: The thoracoscopic approach for instrumentation of scoliosis has advantages of reduced chest wall morbidity compared with the open thoracotomy method but allows comparable curve correction. PMID- 14560201 TI - An innovative technique of vertebral body stapling for the treatment of patients with adolescent idiopathic scoliosis: a feasibility, safety, and utility study. AB - STUDY DESIGN: Retrospective review. OBJECTIVES: To report the feasibility, safety, and utility of vertebral body stapling without fusion as an alternative treatment for adolescent idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: The success rate of brace treatment of adolescent idiopathic scoliosis ranges from 50% to 82%. However, poor self-image and brace compliance are issues for the patient. An alternative method of treatment such as a motion-preserving vertebral body stapling to provide curve stability would be desirable. METHODS: We retrospectively reviewed 21 patients (27 curves) with adolescent idiopathic scoliosis treated with vertebral body stapling. Patients were immature as defined by Risser sign or=6 degrees or beyond 50 degrees was considered a failure of treatment. Of these 10 patients, 6 (60%) remained stable or improved and 4 (40%) progressed. One of 10 (10%) in the stapling group had progressed beyond 50 degrees and went on to fusion. Six patients required stapling of a second curve, three as part of the primary surgery, and three as a second stage, because a second untreated curve progressed. The results need to be considered with caution, as the follow-up is still short. CONCLUSIONS: The data demonstrate that vertebral body stapling for the treatment of scoliosis in the adolescent was feasible and safe in this group of 21 patients. In the short-term, stapling appears to have utility in stabilizing curves of progressive adolescent idiopathic scoliosis. PMID- 14560202 TI - The feasibility, safety, and utility of vertebral wedge osteotomies for the fusionless treatment of paralytic scoliosis. AB - STUDY DESIGN: Before-after intervention study of a fusionless surgical technique to correct scoliosis secondary to spinal cord injury or myelodysplasia in children and adolescents. OBJECTIVES: To determine the feasibility, safety, and utility of a fusionless treatment option for paralytic scoliosis. Once determined, these data could then be applied to develop the application of this operation for patients with other types of scoliosis, such as idiopathic. SUMMARY OF BACKGROUND DATA: The optimal operative treatment for paralytic scoliosis remains to be determined. An ideal procedure would correct the deformity and stop the progression of scoliosis while maintaining mobility of the spine. This latter fact is important, especially for patients who rely heavily on use of trunk mobility for function. METHODS: Fourteen patients with scoliosis secondary to spinal cord injury or myelodysplasia underwent a fusionless vertebral body wedge osteotomy procedure. Feasibility was analyzed by the ability to correct the scoliosis with the osteotomies and preserve mobility. Safety was reported by estimated blood loss, neurologic stability, and complications. Utility was reported by radiographic evidence of arrested curve progression and maintenance of spinal mobility. RESULTS: All 14 patients successfully underwent surgery to insert the wedge-rod system, with an average initial correction of 86% (range 66% 108%). The average estimated blood loss was 1050 cc (range 300-2000 cc). There were no major complications, and no changes in spasticity, bowel or bladder patterns, or motor/sensory levels. There was no case of nonunion at the osteotomy sites. At mean follow-up of 15 months (6-29 months), 10 patients had an improvement in their Cobb magnitude, 1 patient was within 5 degrees of their initial curve, 1 patient had a worse Cobb magnitude, and in 2 patients, the curve direction reversed but still measured less than the preoperative Cobb measurement. Spinal mobility was retained in all patients, as demonstrated on side-bending radiographs. CONCLUSIONS: The vertebral wedge osteotomy procedure appears to be a potential option for the treatment of paralytic scoliosis. The procedure was feasible and safely performed in these 14 patients, with spinal mobility maintained. There were no nonunions. The efficacy of the procedure is still not known, as is for which patients the procedure is indicated and timing of the operation. Long-term follow-up (to skeletal maturity) is needed. Only six of the patients are currently skeletally mature, and more numbers are needed to determine efficacy in this group. PMID- 14560203 TI - Preclinical testing of a wedge-rod system for fusionless correction of scoliosis. AB - STUDY DESIGN: Biomechanical study. OBJECTIVES: This paper describes the biomechanical comparison of calf spines instrumented with a wedge alone versus a wedge-rod construct for the fusionless correction of scoliosis. SUMMARY OF BACKGROUND DATA: Current techniques for the correction of scoliosis require either anterior or posterior spinal fusion for correction. We propose a technique allowing correction via vertebral body osteotomies along with fixation using a wedge-rod construct without the requirement of intervertebral segment fusion. METHODS: Calf spines were used to test the biomechanical difference between the intact spine, transverse osteotomized spine with wedge-rod reconstruction, and transverse osteotomized spine with wedge alone reconstruction. Unconstrained segments (L1-L5) were first tested under five nondestructive static loading conditions to evaluate the intact stability of the operative motion segments in axial compression (-600 N), axial rotation (+/-5.0 Nm, 150 axial preload), flexion and extension (+/-5.0 Nm), and lateral bending (+/-5.0 Nm). Following the intact analysis, vertebral wedge osteotomies in the transverse plane were performed at the L2, L3, and L4 levels. The defects were reconstructed using the Sofamor Danek Wedge Spacer, and stainless steel TSRH one-quarter inch single rod with modified CD HORIZON 6.5-mm diameter vertebral body screws at each level (L2 L4). Standard CD HORIZON 6.5-mm bone screws and staples were used at the superior and inferior ends of the five-level construct. The wedge was on the left side and the rod and screw heads on the right side. After testing the reconstructed specimen, the TSRH rod was removed and the construct retested to evaluate the stability of the wedge alone reconstruction. RESULTS: Construct stiffness was calculated as the peak applied load (N or Nm) divided by the corresponding segmental displacement (mm or degrees) normalized to the intact specimen. Reconstruction static data are expressed as a percentage change from the intact condition. Statistical analysis included descriptives, a one-way analysis of variance, and the Student-Newman-Keuls test for multiple comparisons among the reconstruction groups. Axial compression: under axial compressive loads, the stiffness of the wedge-rod construct was approximately equal to that of the intact group. The stiffness of the wedge alone construct was 56% less than that of the intact group except for extension and left lateral bending. For the other modes of loading (right rotation, left rotation, flexion, extension, right lateral bending, and left lateral bending), the wedge-rod construct was stiffer than that of the intact group. The stiffness of the wedge alone construct was consistently less than that of the intact group. CONCLUSIONS: Based on the results of this biomechanical comparison, the calf spines instrumented with the wedge-rod system for fusionless correction were significantly stiffer as compared to the intact calf spine. The wedges alone were not as stiff as the intact spine. This suggests that the theory of performing transverse osteotomies of vertebral bodies with fixation with wedge-rod construct for 8 to 12 weeks, followed by removal of the rod, could provide adequate fixation and correction of a scoliotic deformity without requiring fusion of motion segments. PMID- 14560204 TI - Tremendous advances continue in the surgical management of spinal deformity. PMID- 14560205 TI - Presidential address: The modern vascular specialist--surgeon, clinician, and interventionist. PMID- 14560206 TI - Does transrenal fixation of aortic endografts impair renal function? AB - OBJECTIVES: Transrenal fixation (TFX) of aortic endografts is thought to increase the risk for renal infarction and impaired renal function. We studied the late effects of TFX on renal function and perfusion. METHODS: Of 189 patients with commercial aortic endografts, which we inserted between 1995 and 2002, we reviewed data for 130 patients (112 men, 18 women) with available creatinine (Cr) concentration and contrast enhanced computed tomography (CT) scans preoperatively and 1 to 97 months after the procedure. Of the 130 patients, 69 patients had TFX and 61 patients had infrarenal fixation (IFX). Both groups were physiologically comparable. Average age was 76 +/- 8 years for patients with TFX and 75 +/- 8 years for patients with IFX. Presence of renal infarct or renal artery occlusion was determined by nephrograms on serial contrast-enhanced CT scans. RESULTS: Mean follow-up was 17 +/- 16 months (range, 1-54 months) for TFX and 21 +/- 21 months (range, 1-97 months) for IFX. Mean serum Cr concentration increased significantly during long-term follow-up in both groups (TFX, 1.3 +/- 0.5 mg/dL to 1.5 +/- 0.8 mg/dL, P <.01; IFX, 1.3 +/- 0.7 mg/dL to 1.4 +/- 0.8 mg/dL, P <.03). Creatinine clearance (CrCl) similarly decreased over long-term follow-up in both groups (TFX, 53.3 +/- 17.7 mL/min/1.73 m(2) to 47.9 +/- 16.2 mL/min/1.73 m(2), P <.01; IFX, 58.1 +/- 22.7 mL/min/1.73 m(2) to 53.1 +/- 23.4 mL/min/1.73 m(2), P <.02). There were no significant differences in the increase in Cr concentration (P =.19) or decrease in CrCl (P =.68) between TFX and IFX groups. Small renal infarcts were noted in four patients (5.8%) in the TFX group and one patient (1.6%) in the IFX group. No increase in Cr concentration or decrease in CrCl was noted in any patient with a renal infarct. Postoperative renal dysfunction developed in 7 of 69 patients (10.1%) in the TFX group and 7 of 61 patients (11.5%) in the IFX group. There were no statistically significant differences between groups with respect to number of patients with new renal infarcts (P =.37) or postoperative renal dysfunction (P =.81). CONCLUSION: There is a slight increase in serum Cr concentration and decrease in CrCl after aortic endografting. However, there was no significant difference in these changes between patients with TFX and IFX. Although TFX may produce a higher incidence of small renal infarcts, these do not impair renal function. Thus our midterm results suggest that TFX can be performed safely, with no greater change in renal function than observed after IFX. PMID- 14560207 TI - Color-flow duplex ultrasound scan versus computed tomographic scan in the surveillance of endovascular aneurysm repair. AB - OBJECTIVE: The purpose of this study is to compare both computed tomographic scan (CT) and color flow duplex ultrasound scanning (CDU) as surveillance modalities for clinically significant endoleaks and to evaluate concordance in abdominal aortic aneurysm (AAA) diameter measurements in patients after endovascular aneurysm repair (EVAR) in a busy hospital vascular laboratory. METHODS: We conducted a retrospective review of all patients who underwent endovascular repair of abdominal aortic aneurysms between February 1996 and November 2002 and had same-day CT and CDU studies. Ninety-seven patients enrolled in phase II clinical studies of Ancure devices had long-term follow-up with both modalities. The other patients underwent simultaneous studies, usually only at the 1-month postoperative visit. Peripheral vascular studies were performed by two certified vascular technicians; all CT scans were reviewed by one vascular surgeon. CT was used as the standard against which the sensitivity, specificity, negative predictive value, and positive predictive value of CDU in endoleak detection was determined. Statistics were performed by using the paired t test; a P value <.05 was considered significant. Kappa statistic was used to assess the correlation between CDU and CT in identifying endoleaks. The correlation between CT and CDU in AAA size measurements as well as in serial size measurements was also determined. RESULTS: Four hundred ninety-five same-day CT and CDU examinations were reviewed in 281 patients. Patients had an average follow-up of 34.6 months (range, 1 to 72 months). Thirty-five leaks were identified among the patients studied (12.4% overall). In comparison with CT, diagnosis of endoleak with ultrasound scanning was associated with a sensitivity of 42.9%, specificity of 96.0%, positive predictive value of 53.9%, and negative predictive value of 93.9%. The correlation between the two modalities was modest (kappa statistic 0.427). The minor axis transverse diameter as measured by ultrasound and CT scans (4.81 +/- 1.1 cm on CT and 4.55 +/- 1.1 cm on ultrasound) correlated closely (r =.93, P <.001.) Seventy percent of paired studies differed by < or =5 mm. Changes in aneurysm size throughout follow-up were -.29 +/-.71 cm on CT scan -.34 +/-.57 cm on duplex ultrasound scan. The correlation coefficient was.65 (P <.001). There was no significant difference in the change as measured by either modality on the paired t test. CONCLUSIONS: Although CDU demonstrates a high degree of correlation with CT scan in determining aneurysm size change over time, it has a low sensitivity and positive predictive value in endoleak detection. In the hospital vascular laboratory at a large tertiary care center, CDU cannot effectively replace CT scan in surveillance after EVAR. PMID- 14560208 TI - Use of cine magnetic resonance angiography in quantifying aneurysm pulsatility associated with endoleak. AB - OBJECTIVE: Persistent aneurysm perfusion or endoleak is associated with pulsatility of abdominal aortic aneurysm (AAA) after endovascular repair. However, the resultant pulsatile change in aneurysm diameter may be difficult to quantify, and therefore its significance is unknown. In this study cine magnetic resonance angiography (MRA) was used to quantify aneurysm wall motion during the cardiac cycle and to correlate it with the presence and type of endoleak. METHODS: Cine MRA was performed in 16 patients undergoing endovascular repair of AAA. A 1.5 T magnet and post-processing with GEMS 4.0 Fiesta computerized video image analysis software were used to calculate maximum aortic diameter during systole and diastole. Changes in aortic diameter were determined from these measurements. Cine MRA was performed on aneurysms before treatment and in patients with and without endoleak after endovascular repair. Type of endoleak was confirmed at angiography in all cases. Four patients had antegrade (type I) endoleak, and eight patients had retrograde (type II) endoleak; no endoleak was present in four patients. Endovascular grafts with stent support throughout the entire length of the graft (Talent) were used in all cases (14 bifurcated grafts, 2 tube grafts). RESULTS: Cine MRA demonstrated significantly greater wall motion and resultant change in aneurysm diameter in patients with type I endoleak compared with patients without endoleak (type I, 2.14 +/- 1.28 mm vs no endoleak, 0.12 +/- 0.09 mm, P =.001). Change in aneurysm diameter in patients with type II endoleak was not significantly greater than in patients with no endoleak (type II, 0.26 +/- 0.21 mm vs no endoleak, 0.12 +/- 0.09 mm, P = NS). Untreated aneurysms demonstrated the greatest change in diameter during the cardiac cycle (3.51 +/- 0.79 mm). CONCLUSION: Cine MRA may be used to accurately quantify AAA wall motion before and after endovascular stent-graft treatment. The extent of change in diameter corresponds to the type of endoleak, with antegrade (type I) endoleak generating greater pulsatile change in diameter than retrograde collateral (type II) endoleak or no endoleak. Cine MRA may provide a noninvasive means of assessing the success of endovascular treatment of AAA. Further studies will be necessary to confirm the utility and efficacy of cine MRA in postoperative assessment of endovascular aneurysm repair. PMID- 14560209 TI - Does hostile neck anatomy preclude successful endovascular aortic aneurysm repair? AB - OBJECTIVES: Poor outcomes have been reported with endovascular aneurysm repair (EVAR) in patients with hostile neck anatomy. Unsupported endografts with active fixation may offer certain advantages in this situation. We compared EVAR results using the Ancure (Guidant) endograft in patients with and without hostile neck anatomy. METHODS: Records of EVAR patients from October 1999 to July 2002 at a tertiary care hospital were retrospectively reviewed from a division database. Patients with elective open abdominal aortic aneurysm (AAA) repair during the same period were reviewed to determine those unsuitable for EVAR. Hostile neck anatomy, assessed by computer tomography (CT) scans and angiograms, was defined as one or more of the following: (1) neck length 3 mm, (3) >2-mm reverse taper within 1 cm below the renal arteries, (4) neck thrombus > or =50% of circumference, and (5) angulation > or =60 degrees within 3 cm below renals. RESULTS: Three hundred and twenty-two patients underwent EVAR with an average follow-up of 18 months. Patients in Phase II trials (n = 41), repaired with other graft types (n = 48), or without complete anatomic records (n = 27) were excluded. Demographics and co-morbidities were similar in the 115 good-neck (GN) and 91 bad-neck (BN) patients except for age (mean, 72.9 years GN vs 75.7 BN; P = 0.13), gender (11% female GN vs 22% BN; P =.04); neck length (mean, 21.8 mm GN vs 14.4 mm BN: P <.001), and angulation (mean, 22 degrees GN vs 40 degrees BN; (P <.001). Perioperative mortality (0 GN vs 1.1% BN), late mortality (5.2% GN vs 4.4% BN), all endoleaks (19.1% GN vs 17.6% BN), proximal endoleaks (0.8% GN vs 2.1% BN), and graft migration (0 for both groups) did not reach statistical significance. Neck anatomy precluded EVAR in 106 of 165 (64%) patients with open AAA. CONCLUSIONS: Unsupported endografts with active fixation can yield excellent results in treating many medically compromised patients with hostile neck anatomy. Nonetheless, an unsuitable neck remains the most frequent cause for open abdominal AAA. PMID- 14560210 TI - Initial successful management of type I endoleak after endovascular aortic aneurysm repair with n-butyl cyanoacrylate adhesive. AB - OBJECTIVE: Transcatheter embolization with coils and other agents has been described as a treatment method for type II endoleak after endovascular aortic aneurysm repair (EVAR). Type I endoleak has not been treated commonly with such therapies, although most investigators believe they warrant definitive intervention. The liquid adhesive n-butyl 2-cyanoacrylate (n-BCA) is often used to treat congenital arteriovenous malformations. The objective of this study is to report our initial experience in treating type I endoleak with n-BCA and with a variety of other interventions. METHODS: A retrospective review was performed of 270 patients who underwent EVAR at our institution between January 1994 and December 2002. Of these, 24 patients had type I endoleak (8.9%), diagnosed either intraoperatively (n = 13, 52%) or during follow-up (n = 12, 48%). Among these 24 patients, 17 had proximal leaks and the remaining 8 patients had distal leaks. These cases form the focus of this study. RESULTS: Twenty-two leaks required endovascular intervention, with the following success rate: n-BCA, 12 of 13 cases (92.3%); extender cuffs, 4 of 5 cases (80%); coils with or without thrombin, 3 of 4 cases (75%). In one patient with persistent endoleak despite attempted endovascular intervention the device ultimately was surgically explanted, and the patient did well. Of six patients with endoleak initially managed expectantly, two eventually underwent attempts at definitive intervention, both with n-BCA. Three sealed spontaneously before definitive intervention could be performed; and in one 97-year-old patient who refused intervention, the aneurysm subsequently ruptured and the patient died. In total, 13 patients with type I endoleak underwent n-BCA transcatheter embolotherapy. No serious complications were directly related to this therapy. Colon ischemia developed in one patient, and was believed to be a result of thromboembolism during wire and catheter manipulation rather than n-BCA treatment. Twelve of these 13 leaks remain sealed at mean follow-up of 5.9 months (range, 0-19 months). CONCLUSION: Our initial use of n-BCA occlusion suggests that it may be an effective and safe method of treatment of type I endoleak after EVAR. In particular, n-BCA embolotherapy may be especially useful in treating type I endoleak not amenable to placement of extender cuffs. Larger case series and longer follow-up are needed before this treatment is more broadly recommended. Type I endoleak after EVAR can be treated successfully with a variety of endovascular methods, and surgical explantation is rarely required. PMID- 14560211 TI - Existence of abdominal aortic aneurysms in patients with thoracic aortic dissections. AB - OBJECTIVE: The objective of this study was to determine the coexistence or later development of pararenal and infrarenal abdominal aortic aneurysms (AAAs) in patients with thoracic aortic dissections. METHODS: One hundred forty-five patients (95 men, 50 women) encountered from 1992 to 2001 with thoracic aortic dissections-excluding those associated with trauma, those with Marfan's syndrome, and those with thoracoabdominal aortic aneurysms-were studied. The most common risk factors included hypertension (59%) and a history of tobacco use (52%). Type III dissections affected 86 patients (59%), and type I dissections affected the remaining 59 patients (41%). Aortic computed tomography (CT) scans were obtained annually. Data were assessed by univariate and multivariate analyses. RESULTS: Five patients (3%) had a history of AAA repair prior to their thoracic aortic dissection diagnosis-3 were type III dissections and 2 were type I dissections. Twelve patient's (8%) AAAs were diagnosed with the initial CT study of their thoracic aortic dissection. Type III dissections accounted for all but one of these (11 of 12, 92%). Ten additional AAAs (7%) developed in the 128 patients with no initial evidence of an AAA being recognized from 1 to 48 months (average 16 months) after the thoracic aortic dissection was diagnosed. Type III dissections affected 8 of these 10 patients. Among the total 27 AAAs noted in this series, 74% (20 AAAs) were not continuous with the thoracic aortic dissection. In the univariate analysis, age (P =.0002), male gender (P =.044), history of smoking (P =.01), chronic obstructive pulmonary disease (P <.001), duration of dissection (P =.05), and presence of type III dissection (P =.009) were associated with the presence of an AAA. In the multivariate analysis, both chronic obstructive pulmonary disease (odds ratio 5.4, 95% CI, 1.3 to 22.3; P =.02) and age (OR 1.06, 95% CI, 1.02 to 1.11; P =.004) were significant predictors of the development of AAAs. CONCLUSION: This study documented that patients with thoracic aortic dissections are at risk to harbor or develop a later AAA. This finding supports the tenet that abdominal CTs or ultrasound scanning should be mandatory in the follow-up of patients with known thoracic aortic dissections. PMID- 14560212 TI - Challenges of endovascular tube graft repair of thoracic aortic aneurysm: midterm follow-up and lessons learned. AB - OBJECTIVES: Endovascular stent-graft repair has great potential in treatment of thoracic aortic aneurysms. This study analyzed a single center's experience with first-generation commercially produced thoracic stent grafts used to treat descending thoracic aortic aneurysms. METHODS: Over 58 months 84 patients underwent endovascular stent-graft repair of descending thoracic aortic aneurysms; 22 patients received the Gore TAG stent graft, and 62 patients received the Talent thoracic endovascular stent-graft system. Each patient was enrolled in one of three distinct US Food and Drug Administration trials at Mount Sinai Medical Center in accordance with strict inclusion and exclusion criteria, including suitability for open surgery, aneurysm anatomy, and presence of comorbid medical illness. Mean age of this cohort was 71 +/- 12 years. There were 54 men and 30 women, and 74 (88%) had three or more comorbid illnesses. Primary technical success was achieved in 76 patients (90%). Mean follow-up was 15 months (range, 0-52 months). RESULTS: Successful aneurysm exclusion was achieved in 69 patients (82%). Major procedure-related or device-related complications occurred in 32 patients (38%). There were six proximal attachment failures (8%), four distal attachment failures (6%), one intergraft failure (1%), two mechanical device failures (3%), five periprocedural deaths (6%), and five late aneurysm ruptures (6%). At 40 months, overall survival was 67% (+/-10%), and freedom from rupture or from type I or type III endoleak was 74% (+/-10%). CONCLUSION: While promising, this midterm experience with commercially available devices highlights the shortcomings of current stent-graft technology. Three significant advancements are required to fulfill the potential of this important treatment method: a stent graft with a durable proximal and distal fixation device, enhanced engineering to accommodate high thoracic aortic fatigue forces, and a mechanism to adapt to aortic arch and visceral segment branches to enable treatment of lesions that extend to or include these vessels. PMID- 14560213 TI - Intraoperative duplex ultrasound of visceral revascularizations: optimizing technical success and outcome. AB - PURPOSE: The purpose of this study was to evaluate the use of intraoperative duplex ultrasound scanning (IOUS) during visceral revascularizations and correlate its results with clinical outcome. METHODS: We studied 68 patients (15 men and 53 women, mean age 66.5 years, range 27-86 years) who underwent visceral revascularization with concomitant IOUS examination of 120 visceral arteries (52 celiac, 60 superior mesenteric, and 8 inferior mesenteric arteries) from 1992 to 2002. Patients were divided into two groups on the basis of ultrasound findings: normal and abnormal IOUS. The incidence of early and late graft-related complications (thrombosis, restenosis, recurrent symptoms, reintervention) and graft-related death was compared in both groups. RESULTS: One-hundred and two (85%) arteries had normal IOUS. Eight (6.6%) arteries had minor defects, including small kinks (4), mild residual stenoses (3), and small intimal flap (1). Ten (8.4%) arteries had major defects, consisting of hemodynamically significant residual stenoses (4), thrombus (2), kinks (2), bidirectional flow (1), and intimal flap (1). Major defects were successfully revised in all except three cases: two persistent mild stenoses and one bidirectional flow. Patients with abnormal IOUS at the end of the operation had increased incidence of graft related complications and/or death (55.5% vs 7.8%; P =.004), early graft thrombosis (14.2% vs 1.0; P =.04), reintervention (21.4% vs 3.2%; P =.03), and graft-related death (33.3% vs 1.9%; P =.02), compared with patients with normal IOUS. CONCLUSION: This study supports the routine use of IOUS during visceral revascularizations to optimize technical success and outcome. Persistent ultrasound scanning abnormalities are associated with risk of early graft failure, reintervention, and death. Patients with normal ultrasound scans can expect excellent results. PMID- 14560214 TI - Endovascular treatment of celiac and mesenteric arteries stenoses: applications and results. AB - PURPOSE: To evaluate the safety and assess the role of endovascular therapy in a variety of conditions related to celiac and mesenteric vascular occlusive disease. Patients and methods Our retrospective study population included 25 consecutive patients (mean age, 66 years), in whom 28 procedures were performed on 26 stenosed or occluded mesenteric vessels (superior mesenteric artery [SMA] or celiac artery [CA]). Indications included chronic mesenteric ischemia (21 patients), including 2 patients who underwent stenting prior to a planned operative repair of a juxtamesenteric AAA. Three liver transplantation patients underwent stenting of an associated CA stenosis. One patient with a splenorenal bypass underwent stenting on an associated CA stenosis. The technical and clinical success rates and the incidence of complications were determined. Follow up parameters included maintained patency on duplex sonography and sustained clinical benefit. The need for additional interventions was noted. RESULTS: All procedures but one were technically successful (96%). Major complications occurred in three patients (one transient contrast-induced nephrotoxicity and two pseudoaneurysms). Immediate clinical success was achieved in 22 patients (88%). The three clinical failures included two patients with an excellent angiographic outcome, but with single-vessel moderate severity disease. Survival table analysis of delayed clinical outcome showed primary and primary-assisted clinical benefits at 11 months of 85% and 91%, respectively. Primary and primary-assisted stent patencies, as assessed by duplex sonography and/or angiography, at 6 months were both 92%. Angiographically documented restenosis occurred in three patients. Restenosis in two patients with CA stents was due to extrinsic compression, and it was without symptoms in one patient and was treated satisfactorily by restenting in the other patient. Restenosis in one patient with an SMA stent was successfully treated by restenting. CONCLUSIONS: Our experience suggests a potential role for endovascular therapy of celiac and mesenteric arterial occlusive disease in a variety of clinical scenarios, with a low incidence of complications and a high technical success rate. PMID- 14560215 TI - Endovascular therapy in prevention and management of coronary-subclavian steal. AB - BACKGROUND: The optimal management of patients undergoing coronary artery bypass grafting (CABG) who have proximal subclavian artery stenosis (SAS) is not well established. SAS may lead to flow reversal through a patent in situ internal mammary artery graft, resulting in myocardial ischemia (coronary-subclavian steal). We review our experience in prevention and management of coronary subclavian steal. METHODS: The medical records of patients who received treatment of symptomatic coronary-subclavian steal were reviewed. Patients who underwent subclavian artery revascularization before CABG were also included in our review. Patient demographic data, findings at presentation, imaging and treatment methods, and short-term and intermediate-term results were analyzed. RESULTS: Over 4 years, 14 patients with combined subclavian and coronary artery disease were identified. Nine patients had angina (n = 8) and/or congestive heart failure (n = 2) after CABG (post-CABG group). Four patients underwent treatment of SAS and one underwent treatment of recurrent stenosis before or during CABG (pre-CABG group). Among this pre-CABG group, one patient had symptoms of left arm claudication; the other four patients had no symptoms. A blood pressure gradient was commonly noted between both arms. An angiogram confirmed the proximal location of SAS in all patients, and established the presence of flow reversal in a patent internal mammary artery graft in the post-CABG group. Operative management consisted of percutaneous transluminal angioplasty (PTA) and stenting of the subclavian lesion in 11 patients, PTA only in 2 patients, and carotid subclavian bypass grafting in 1 patient. No known perioperative complications or morbidity was encountered in either group. Mean follow-up was 29 months, during which stenosis recurred in two patients, along with associated cardiac symptoms. In both patients repeat angioplasty was successful, for an assisted primary patency rate of 100%. CONCLUSION: PTA and stenting to treat SAS appears to provide effective protection from and treatment of coronary-subclavian steal over the short and intermediate terms. A surveillance program is essential because of the risk for recurrent stenosis. Continued follow-up is necessary to determine long-term efficacy of this treatment compared with more conventional surgical approaches. PMID- 14560217 TI - A model for predicting occult carotid artery stenosis: screening is justified in a selected population. AB - OBJECTIVES: The diagnosis and treatment of carotid artery disease is an integral part of stroke prevention. However, a population of patients who would benefit from screening for carotid artery stenosis has not been well defined. As part of an institutional stroke-screening program, a modified, rapid duplex scan was developed to evaluate patients for occult carotid stenosis. The goal of this study was to evaluate risk factors predictive of carotid stenosis in a selected population, and to identify patients who would benefit from carotid screening. METHODS: Patients were eligible for the study if they were >60 years of age and had a history of hypertension, heart disease, current smoking, or family history of stroke. A modified carotid duplex scan that had been previously validated against formal duplex scanning was utilized; this involved visualization of the carotid bulb and proximal internal carotid artery where Doppler flow velocities were obtained and recorded. RESULTS: Screening was performed on 394 patients. Thirty-eight patients (9.6%) had either unilateral or bilateral carotid stenosis of > or =50%. Risk factors evaluated included smoking, hypertension, cardiac disease, or hypercholesterolemia. If none of these risk factors was present, the incidence of carotid stenosis was 1.8%. This increased to 5.8% with one risk factor, 13.5% with two risk factors, and 16.7% with three risk factors. Two of three patients with all four risk factors had carotid stenosis (66.7%). Logistic regression and prespecified contrast statements for multiple comparisons were used to assess the relationship between the presence of risk factors and occult carotid artery stenosis. The presence of any one of these risk factors was associated with a statistically significant increase in the presence of occult carotid stenosis (P <.01). This was also statistically significant for the presence of any two risk factors (P <.01) or three risk factors (P <.05). CONCLUSION: The prevalence of carotid stenosis significantly increases with the presence of one or more identifiable demographic risk factors in a selected population. Assuming the diagnosis and treatment of carotid stenosis are fundamental to stroke prevention, screening for carotid artery disease is justified in this group of patients. PMID- 14560218 TI - Managing PAD with multiple platelet inhibitors: the effect of combination therapy on bleeding time. AB - PURPOSE: Patients with lower-extremity peripheral arterial disease (PAD) face a high risk of cardiovascular morbidity and mortality. Platelet inhibition (PI) significantly reduces this risk. Combination PI is common and increasingly indicated in patients with PAD; however, the effect on platelet function has not been objectively evaluated. Aspirin (ASA), clopidogrel (Clop), and cilostazol (Cilo) are the three most commonly used PI drugs in patients with PAD. A prospective, sequential evaluation of platelet function using the template bleeding time (BT) was performed for PAD patients taking these medications singly and in combination. METHODS: Twenty-one patients with PAD, averaging 65.9 years of age, were studied. Patients were placed on sequential two-week regimens of the following therapies: washout (no PI), ASA (325 mg daily), ASA + Cilo (100 mg twice daily), washout, Cilo, Cilo + Clop (75 mg each day), washout, Clop, Clop + ASA, and Clop + ASA + Cilo. At the end of each phase, trained personnel measured the BT. RESULTS: Baseline bleeding time for the group was 4.29 +/- 1.69 minutes. ASA (BT = 6.64 +/- 3.52) and Clop (BT = 10.17 +/- 5.4) significantly prolonged bleeding time (P < 0.01); however, no significant effect was observed with Cilo alone (BT = 5.41 +/- 2.69). Combined treatment with ASA + Clop (BT = 17.39 +/- 4.59) had a more pronounced effect on BT compared with either agent alone (P < 0.01). The addition of Cilo to either ASA (BT = 8.3 +/- 4.27) or Clop (BT = 12.7 +/- 7.46) or the combination of ASA + Clop (BT = 17.92 +/- 4.69) did not prolong BT. CONCLUSION: All patients with PAD require platelet inhibition, and many require pharmacotherapy for intermittent claudication. The platelet inhibitors aspirin and clopidogrel are used for the reduction of ischemic events. They significantly prolong bleeding time individually and to a greater extent in combination. Cilo is used to improve walking distance in patients with intermittent claudication. When Cilo is added to ASA, Clop, or the combination of the two, there is no additional increase in bleeding time. Therefore, Cilo can be used in combination with other platelet inhibitors without an additional effect on platelet function as reflected by the bleeding time. PMID- 14560219 TI - The LifeSite Hemodialysis Access System in patients with limited access. AB - OBJECTIVE: The LifeSite Hemodialysis Access System was recently introduced as a completely subcutaneous device with reported advantages of improved patient comfort and reduced catheter-related infection. The performance of the LifeSite catheter at a single, tertiary-care university medical center was reviewed. METHODS: We retrospectively reviewed all patients who underwent placement of the LifeSite catheter between February 2001 and March 2002. Kaplan-Meier analysis was used to determine the probability of patient survival, freedom from catheter related infection, and freedom from device failure necessitating catheter removal. RESULTS: Thirty-six patients who had previously received dialysis for an average of 6.1 years underwent placement of 37 LifeSite catheters. Most patients (95%) were referred for LifeSite placement because they had exhausted all available arteriovenous fistula and graft sites. Mean follow-up was 6.8 months, with a patient survival rate of 81% at 8 months. Primary and secondary patency rates were 62% and 87% at 8 months, respectively. Two patients died from infectious device-related complications. Twelve of 17 patients (71%) with device related infection did not manifest any signs or symptoms at the valve site. There were 2.4 catheter-related infections and 2.6 device failures requiring removal per 1000 patient-catheter days. Freedom from infection and device removal at 8 months was 46% and 49%, respectively. CONCLUSIONS: The LifeSite demonstrated acceptable patency, infection, and device failure rates; however, in patients with limited access, unrecognized infection and death may occur. The LifeSite should not be used as a substitute for a more permanent form of hemodialysis access. PMID- 14560220 TI - Salvaging prosthetic dialysis fistulas with stents: forearm versus upper arm grafts. AB - OBJECTIVE: We compared results of angioplasty with those of concomitant stent placement to treat thrombosed forearm hemodialysis grafts with results for upper arm grafts. MATERIAL AND METHODS: Between October 1998 and July 2002, stents were deployed in 61 patients undergoing balloon angioplasty because of venous anastomotic stenosis causing graft thrombosis. Stents were used only in cases of inadequate angioplasty results. Twenty-three forearm grafts and 38 upper arm grafts were treated. All procedures were performed in an endovascular operating suite, with fistulography. Primary and secondary patency rates were analyzed and compared for graft location with the life table method. RESULTS: Grafts had undergone a mean of 1.56 previous revisions because of thrombosis (forearm: 1.52, upper arm: 1.58; P = NS). Excluding early thrombosis, a single graft infection was the only procedural complication. Cumulative primary patency rate at 3, 6, and 12 months (from stent placement) was 36.4%, 15.6%, and 0%, respectively, for forearm grafts, which was inferior to the 59.5%, 34.0%, and 17.0% primary patency rate observed for upper arm grafts (P =.0307) Secondary patency rate was 40.9%, 40.9%, and 30.7%, respectively, for forearm grafts, and 64.9%, 42.3%, and 19.7% for upper arm grafts (P = NS). CONCLUSION: Stent deployment can salvage thrombosed dialysis grafts. However, sustained patency occurs infrequently, with better results for upper arm grafts than for forearm grafts. Inasmuch as surgical revision of forearm grafts is usually straightforward, stenting should be reserved for use in high axillary grafts and other sites where surgical repair is difficult. PMID- 14560221 TI - Tissue (muscle) oxygen saturation (StO2): a new measure of symptomatic lower extremity arterial disease. AB - OBJECTIVES: Near-infrared spectroscopy provides a noninvasive method of measuring tissue oxygen saturation and has been used to monitor extremity compartment syndrome. Tissue O(2) saturation (StO(2)) is potentially useful in assessing patients with peripheral arterial disease (PAD). The purposes of this feasibility study are to (1) explore the diagnostic sensitivity of StO(2) in subjects with PAD and symptoms of intermittent claudication (IC) compared with normal subjects, and (2) correlate the change in StO(2) during and after exercise with the ankle brachial index (ABI) in patients with IC. Material and methods Forty-nine subjects, 35 normal and 14 PAD, from two centers were evaluated in a prospective cross-sectional analysis comparing StO(2) by using the InSpectra tissue spectrometer and ABI at rest (baseline) and after treadmill exercise. Measurements were obtained at baseline and peak exercise (normal subjects) and at baseline, initial claudication distance (ICD) and absolute claudication distance (ACD) in PAD subjects. Endpoint values were the mean of 15 data points. Times to 50% of StO(2) recovery to baseline (T(50)) and complete recovery to baseline (T(100)) were measured. Receiver-operator characteristic curves were constructed to assess the sensitivity/specificity values associated with various StO(2) cut points. RESULTS: The PAD patients were older (P =.0002) and 57% were male, compared with 37% males in the normal group. The ABI was 0.68 +/- 0.14 in PAD patients versus 1.14 +/- 0.08 in normal subjects (P <.0001). The baseline StO(2) was 65% in both groups. The peak exercise StO(2) was significantly lower and the absolute change in StO(2) and the percent change in StO(2) were significantly greater in PAD patients (P < 0.45). The T(50) and T(100) were longer in the PAD patients compared to normal subjects (P =.0001 and.002, respectively). A T(50) of >70 seconds yielded a sensitivity of 89% and a specificity of 85% for PAD. CONCLUSIONS: StO(2) is a new and potentially useful technique to evaluate patients with PAD. Resting StO(2) was similar in PAD-IC subjects and normals. There was a significantly greater drop in StO(2) and longer recovery times in PAD IC subjects. Interestingly, StO(2) at the ICD and ACD was similar. StO(2) offers a different and perhaps more appropriate end point for diagnosis and monitoring of the management of patients with PAD, and may offer additional insight into the pathophysiology of exercise-induced muscle ischemia and its recovery. PMID- 14560222 TI - Endovascular repair of abdominal aortic aneurysm without preoperative arteriography. AB - OBJECTIVE: Clinical trials of endovascular aortic aneurysm repair (EVAR) have required both preoperative aortography and computed tomography (CT). We codeveloped specialized three-dimensional (3-D) reconstruction and computer-aided measurement, planning, and simulation software (3-D CAMPS) based on CT or magnetic resonance imaging, to eliminate the need for preoperative arteriography. METHODS: EVAR with 3-D CAMPS as the sole preoperative imaging method was performed in 196 patients from 1996 to 2001, with six endograft types in three configurations. Physical examination, abdominal radiography, and CT (3D-CAMPS) were performed at 1, 6, and 12 months, then annually. RESULTS: For a subset of cases in which a comparison could be made, 3-D CAMPS was superior to angiography for prediction of endograft length and iliac access. Hospital mortality was zero, and 30-day mortality was 0.5%. In three patients immediate conversion to open repair (1.5%) was necessary because of previously unknown stent-graft mechanical limits. Incidence of endoleak was 15% at 1 month, 10% at 6 months, 6% at 12 months, and 7% at 24 months, and 92% of endoleaks were type II. Mean follow-up was 18 months. Aneurysm-related mortality was zero. Nineteen secondary procedures (all endovascular) were performed in 16 patients (8%). For all graft types, freedom from secondary procedure was 94% at 1 year and 90% at 2 years, and this was better for endografts ultimately approved by the US Food and Drug Administration (96% at 1 year, 95% at 2 years; P =.02). No known measurement related complications occurred in the series. Results for secondary intervention and endoleak compare favorably to series with similar endograft types. CONCLUSIONS: EVAR can be performed with 3-D CAMPS as the sole preoperative imaging method to achieve outcomes comparable to the best series published for each endograft type. CT with 3-D CAMPS can effectively eliminate the need for preoperative arteriography and avert associated morbidity, expense, and exposure to contrast agent and radiation. PMID- 14560223 TI - Surgeon specialty and provider volumes are related to outcome of intact abdominal aortic aneurysm repair in the United States. AB - OBJECTIVE: This study was undertaken to determine the relative importance of surgeon specialty, hospital volume, and surgeon volume on outcome after abdominal aortic aneurysm (AAA) repair. METHODS: Data were reviewed for 3912 patients undergoing AAA repair in the Nationwide Inpatient Sample during 1997. In-hospital mortality was compared between high-volume hospitals and low-volume hospitals and between high-volume surgeons and low-volume surgeons. High-volume hospitals performed more than 35 AAA repairs per year, and high-volume surgeons performed more than 10 AAA repairs per year. Vascular, cardiac, and general surgery specialization was identified by analysis of other procedures performed by each surgeon. RESULTS: Overall, AAA repair mortality was 4.2%, and was lower at high volume hospitals (3.0%) than at low-volume hospitals (5.5%) (P <.001). Lowest mortality was associated with operations performed by vascular surgeons (2.2%) compared with cardiac surgeons (4.0%) and general surgeons (5.5%) (P <.001). Mortality rates were also lower for high-volume hospitals (2.5%) compared with low-volume hospitals (5.6%) (P <.001). In a risk-adjusted analysis, high-volume hospital, vascular surgery specialty, and high-volume surgeon were all independently associated with lower risk of in-hospital mortality. In this analysis, risk reduction was 30% for high-volume hospitals (95% confidence interval [CI], 2%-51%; P <.05) and 40% for surgery by a high-volume surgeon (95% CI, 12%-60%; P =.01). AAA repair by general surgeons compared with vascular surgeons was associated with 76% greater risk for death (95% CI, 10%-190%; P =.02). No significant difference in mortality was found between cardiac and vascular surgeons. CONCLUSIONS: High surgeon volume and hospital volume of AAA repair were both associated with lower mortality compared with low-volume providers. Increased specialization in vascular surgery was associated with markedly decreased mortality independent of AAA repair volume. Health policy in support of selective referral for AAA repair should consider surgical specialization in addition to provider volume thresholds. PMID- 14560225 TI - Is cross-femoral bypass grafting a disadvantage of aortomonoiliac endovascular aortic aneurysm repair? AB - PURPOSE: The need for cross-femoral bypass grafting (CFBG) is considered by some to be a major disadvantage of endovascular aneurysm repair (EVAR) with the aortomonoiliac technique. To determine the durability of CFBG in this setting, we examined data from 148 consecutive high-risk patients in a clinical trial of EVAR with a custom-made aortomonoiliac endovascular stent graft. METHODS: All data were collected prospectively. After hospital discharge, patients were evaluated at 1, 3, and 6 months and annually thereafter. All CFBG was constructed of expandable polytetrafluoroethylene. RESULTS: During follow-up averaging 23.6 +/- 16.2 months, nine CFBG complications developed in 8 patients (5.4%), including disruption (n = 2), infection (n = 3), thrombosis (n = 2), and pseudoaneurysm (n = 3). Four patients with CFBG complications died, of consequences of infection (n = 2), intracranial hemorrhage during attempted CFBG thrombolysis (n = 1), and intracranial hemorrhage during anticoagulation (n = 1). There were no amputations. At life table analysis, freedom from CFBG complication was 96.3% +/- 1.6% at 12 months, 94.1% +/- 2.2% at 24, 36, and 48 months, and 86.2% +/- 7.8% at 60 months. Overall survival for this high-risk patient group was 83.4% +/- 3.1% at 12 months, 70.4% +/- 4.1% at 24 months, 56.5% +/- 5.3% at 36 months, and 44.8% +/- 6.4% at 48 months. CONCLUSION: CFBG is durable, with a low rate of complications in patients undergoing aortomonoiliac EVAR. Need for CFBG should not discourage use of aortomonoiliac devices in patients with anatomy unfavorable for other EVAR approaches. PMID- 14560224 TI - Quality of life, impotence, and activity level in a randomized trial of immediate repair versus surveillance of small abdominal aortic aneurysm. AB - OBJECTIVE: We compared long-term health-related quality-of-life outcome after randomization to immediate elective repair or imaging surveillance, and in relation to time of elective repair, in patients with small asymptomatic abdominal aortic aneurysm (AAA). METHODS: This randomized clinical trial was carried out in 16 Veterans Affairs medical centers. Study subjects were patients at good surgical risk, aged 50 to 79 years, with AAAs 4.0 to 5.4 cm in diameter. Interventions included immediate open surgical AAA repair or imaging surveillance every 6 months with repair reserved for AAAs that became symptomatic or enlarged to 5.5 cm. Main outcome measures considered were SF-36 health status questionnaire, prevalence of impotence, and maximum activity level, which were determined at randomization and at all follow-up visits. RESULTS: Eleven hundred thirty-six patients were randomized and followed up for 3.5 to 8 years (mean, 4.9 years). The two randomized groups did not differ significantly for most SF-36 scales at most times, but the immediate repair group scored higher overall in general health (P <.0001), which was particularly evident in the first 2 years after randomization, and slightly lower in vitality (P <.05). The baseline value of one SF-36 scale, physical functioning, was an independent predictor of mortality. Overall, more patients became impotent after randomization to immediate repair compared with surveillance (P <.03), but this difference did not become apparent until more than 1 year after randomization. Maximum activity level did not differ significantly between the two randomized groups, but decline over time was significantly greater in the immediate repair group (P <.02). CONCLUSIONS: For most quality-of-life measures and times there was no difference between randomized groups. Immediate repair resulted in a higher prevalence of impotence more than 1 year after randomization, but was also associated with improved perception of general health in the first 2 years. PMID- 14560226 TI - Endovascular aneurysm repair with a bifurcated endovascular graft at a primary referral center: influence of experience, age, gender, and aneurysm size on suitability. AB - OBJECTIVE: The purpose of this study was to assess the suitability for endovascular repair of abdominal aortic aneurysm (EVAR) in an unselected patient population. PATIENTS AND METHODS: Between February 1999 and May 2002 all consecutive patients with a nonemergent abdominal aortic aneurysm (AAA) were prospectively examined with contrast material-enhanced spiral computed tomography (CT). Those patients probably suitable for EVAR on the basis of CT findings underwent calibrated angiography. A panel of radiologists and vascular surgeons reviewed the clinical data and vascular anatomy, and decided on the appropriateness of EVAR with the bifurcated Zenith AAA endovascular graft. RESULTS: One hundred seven patients were included. Fifty-six patients (52%) had one or more contraindications for EVAR. Unsuitability was most frequently (88%) related to the proximal neck. Inadequate neck length was the most common specific reason. Inadequate iliac anatomy was the reason for unsuitability in 59% of patients. The rate of unsuitability decreased from 61% during the first half of the study to 40% during the second half (P =.03) Unsuitability was equal between men and women. Age and maximum diameter did not differ between candidates and noncandidates. CONCLUSION: Almost half (48%) of patients with an infrarenal AAA referred to a primary referral center are suitable for EVAR with the bifurcated Zenith AAA endovascular graft. Neck anatomy was the most frequent reason for rejection. Rate of suitability increased over time, probably as a result of increasing experience. Suitability was not influenced by gender, age, or aneurysm size. PMID- 14560227 TI - Selective management of abdominal aortic aneurysms smaller than 5.0 cm in a prospective sizing program with gender-specific analysis. AB - PURPOSE: We present extended follow-up findings of the Kingston prospective sizing program for patients with abdominal aortic aneurysm (AAA) smaller than 5.0 cm in diameter, with gender-specific analysis. METHODS: From 1976 to 2001, 895 patients (688 men, 207 women) with AAA smaller than 5.0 cm were entered, regardless of fitness, in a prospective sizing program in which computed tomography scans were obtained every 6 months. Operations were performed in fit patients with an increase in AAA size to 5 cm (n = 190), AAA expansion greater than 0.5 cm in 6 months (n = 27), or for other reasons (n = 33). Follow-up continued until AAA rupture, surgery, death, or removal from the program. RESULTS: No AAA smaller than 5.0 cm ruptured during prospective follow-up. There was a statistically significant increase in expansion rate relative to size at entry, with the highest mean expansion rate of 0.52 cm/y for AAA 4.5 to 4.9 cm in diameter. There was no significant difference in AAA expansion rate between men and women. The frequency of surgery was inversely related to age at entry, but was positively related to AAA size at entry, with patients with AAA 4.5 to 4.9 cm at entry 6.8 times more likely (95% confidence interval, 4.3-10.7) to undergo surgery than those with AAA 3.0 to 3.4 cm at entry. Women were older than men at entry, and age at entry in those undergoing surgery was significantly greater in women. CONCLUSIONS: The study confirms the results of the United Kingdom Small Aneurysm Trial and the Aneurysm Detection and Management Study, that is, that risk for rupture is extremely unlikely with AAA smaller than 5.0 cm, which enables safe follow-up surveillance programs in both men and women with AAA smaller than 5.0 cm. PMID- 14560228 TI - Prospective randomized multicenter trial of fibrin sealant versus thrombin-soaked gelatin sponge for suture- or needle-hole bleeding from polytetrafluoroethylene femoral artery grafts. AB - OBJECTIVE: We evaluated the safety and efficacy of the fibrin sealant Beriplast P (FSBP; Aventis-Behring) for hemostasis in anastomosis of polytetrafluoroethylene (PTFE) grafts to the femoral artery. METHODS: In a single-blinded randomized prospective multicenter clinical trial, FSBP was compared with thrombin-soaked gelatin sponge (TSG) for efficacy in stopping bleeding from needle or suture holes in PTFE grafts after anastomosis to the femoral artery. Patients were randomized to FSBP application, which requires a 3-minute period of arterial clamping to enable the fibrin clot to adhere, or to TSG application, which requires pressure from gauze sponges, after completion of the femoral artery anastomosis. The primary end point was hemostasis, defined as absence of any detectable bleeding as judged by the operating surgeon, by 4 minutes after randomization. Secondary end points included actual time from randomization to hemostasis, time to beginning of wound closure, measured blood loss (weighed sponges), incidence of recurrent bleeding, stay in the intensive care unit, and hospital length of stay. Data were analyzed with the intention-to-treat method. RESULTS: Two hundred thirty-five subjects were enrolled at 26 medical centers; 34 were subsequently excluded from the study. Of the 201 randomized subjects, 100 received FSBP and 99 received TSG. Hemostasis was achieved by 4 minutes in 64 subjects (63%) in the FSBP group and 40 subjects (40%) in the TSG group (P =.0018). In the FSBP group, compared with the TSG group, time to hemostasis was shorter (median, 4.0 minutes; 95% confidence interval [CI], 3.8-4.18 minutes vs median, 5.6 minutes, 95% CI, 4.5-7.0; P =.008), blood loss was less (mean, 4.0 +/ 29.7 g vs mean, 15.6 +/- 28.4 g; P <.0001), and time to wound closure was shorter (median, 15 minutes; 95% CI, 10.47-18.67 minutes vs median, 22.8 minutes; 95% CI, 18.67-30.67; P =.005). There were no differences in recurrent bleeding or any other adverse events. There was no significant difference in ICU stay, but hospital length of stay was shorter in the FSBP group compared with the TSG group, and the difference approached significance (median, 6.5 days; 95% CI, 5.00 7.00 days vs median, 7.0 days; 95% CI,. 6.00-8.00 days; P =.0565). CONCLUSION: FSBP is more effective than TSG for achieving hemostasis of needle or suture hole bleeding from PTFE femoral artery grafts. PMID- 14560229 TI - Hyperhomocysteinemia and risk for peripheral arterial occlusive disease in young women. AB - OBJECTIVE: Few studies to date have examined the relationship between hyperhomocysteinemia and peripheral arterial occlusive disease (PAOD) in young women. In this study we assessed hyperhomocysteinemia as a risk factor for PAOD in young women. In addition, we evaluated the effect of joint exposure to hyperhomocysteinemia and traditional risk factors. METHODS: Two hundred twenty women, ages 18 to 49 years, with PAOD and 629 healthy women (control group) from a population-based case-control study filled out the same structured questionnaire and donated venous blood samples for determination of plasma homocysteine levels. Hyperhomocysteinemia was defined as nonfasting total plasma homocysteine level above the 90th percentile of the control range. RESULTS: Young women with hyperhomocysteinemia had a 2.5-fold (95% confidence interval [CI], 1.7 3.9) increased risk for PAOD. When presence of hyperhomocysteinemia was combined with presence of a traditional risk factor, relative risk strongly increased in smokers (odds ratio [OR], 18.9; 95% CI, 8.3-42.9) and in women with hypertension (OR, 10.3; 95% CI, 5.4-19.8), hypercholesterolemia (OR, 8.5; 95% CI, 4.2-17.1), and diabetes (OR, 8.9; 95% CI, 1.7-46.9). CONCLUSIONS: Hyperhomocysteinemia is a risk factor for PAOD in young women. There is a strong synergistic effect between hyperhomocysteinemia and all traditional vascular risk factors. Our findings may have implications for risk management in these young women. PMID- 14560230 TI - Variation and changes in state-specific carotid endarterectomy and 30-day mortality rates, United States, 1991-2000. AB - OBJECTIVES: The objectives of this study were to investigate variations between states and changes in state-specific carotid endarterectomy (CEA) and 30-day mortality rates. Cross-sectional variations and changes over time in such measures may be indicative of improvement in the quality of care. METHODS: We performed retrospective analyses of pre-existing administrative data on Medicare beneficiaries aged 65 years and older in the United States. Age-adjusted, state specific CEA rates and 30-day postoperative mortality rates in 1991, 1995 and 2000 were examined, as well as changes in these rates from 1991 to 1995 and from 1995 to 2000. Stroke mortality in the general population of each state was used as a crude measure of the need for CEA procedure in the state. The Spearman rank correlation analysis was used to study correlations between rates. Oldham's method was used to avoid the effect of regression to the mean. RESULTS: There were wide variations in the state-specific CEA rates, 30-day mortality, and in changes in these rates over time. The states with relatively low procedure rates in 1991 also had low rates in 1995 and 2000, and relatively higher increases in the rates. The states with relatively high 30-day mortality in 1991 or 1995 had lower increases or greater decreases in the rate. CEA rates were not correlated with any measure of surgical mortality, but they were correlated with stroke mortality in the general population. CONCLUSIONS: The inter-state variation in CEA rates has not changed much since 1991, but variation in 30-day mortality decreased through 2000. The states with low procedure rates in 1991 did not have sufficient increase to catch up with the high-rate states by 1995, but they were prone to experience a higher increase in the subsequent 5 years. The validity of stroke mortality in a state as a measure of the need for CEA is questionable. Further research using clinical data is needed to better explain variations between states. PMID- 14560231 TI - Theoretical hydraulic consequences of vein graft taper. AB - OBJECTIVE: Internal diameter is a strong predictor of patency of infrainguinal vein grafts. However, most vein grafts are tapered, with variable diameter along their length. It is unknown which diameter is most important in determining graft resistive properties, that is, its mean diameter, minimum diameter, or some geometric combination thereof. The purpose of this analysis was to examine the hydraulic consequences of vein graft tapering, with longitudinal impedance (Z(L)), a conduit-specific measure of pulsatile resistance along straight rigid tubes. METHODS: Proximal and distal graft pressure, pressure gradient (DeltaP), and blood flow (Q) were measured intraoperatively in a 100 cm bypass graft and digitally recorded for 10 seconds at 200 Hz. With the Womersley solution for fully developed fluid flow in a rigid tube, a series of DeltaP waveforms were generated for graft diameters ranging from 1.2 to 8.2 mm. With an axisymmetric form of the Navier-Stokes equations, a second series of DeltaP waveforms were computed for grafts with long smooth symmetric tapers ranging from 0% to 90%, with geometric mean diameter of 3.2, 4.2, and 5.2 mm (%Taper = 100 x [proximal diameter - distal diameter]/proximal diameter). For each set of DeltaP and Q, Z(L) was calculated as DeltaP/Q, plotted over a range of 8 Hz, and integrated over 4 Hz to yield integral Z(L). RESULTS: The architecture of the calculated DeltaP and Z(L) waveforms closely approximated their measured counterparts, validating the method. As expected, Z(L) was highly diameter-dependent in a nonlinear fashion. With a clinically relevant boundary of less than 50 x 10(3) dyne/cm(5) as "acceptable," the minimum acceptable diameter of nontapered 100 cm bypass conduits was 4.3 mm. Analysis of graft taper revealed that small amounts of taper in large conduits were well-tolerated. For example, introduction of 32% taper in a 5.2 mm graft (6.2 mm --> 4.2 mm) caused only an 8% increase in integral Z(L) (from 32 to 35 x 10(3) dyne/cm(5)). More pronounced taper in smaller conduits rendered them unacceptable. For example, 53% taper of a 4.2 mm graft (5.7 mm --> 2.7 mm) created a conduit with integral Z(L) of 70 x 10(3) dyne/cm(5), well above the acceptable limit. The relationship between Z(L) and percent taper was nonlinear and strongly dependent on mean diameter. CONCLUSIONS: The relationship between Z(L) and diameter in vein grafts is nonlinear; thus Z(L) increases rapidly in conduits smaller than 4 mm. Tapered vein grafts behave hydraulically like nontapered grafts, provided their geometric mean is greater than 4 mm and their degree of taper is less than 40%. Tapered veins are satisfactory conduits for long-segment bypass grafts, provided their mean diameter is acceptable. PMID- 14560232 TI - Definition of venous reflux in lower-extremity veins. AB - PURPOSE: This prospective study was designed to determine the upper limits of normal for duration and maximum velocity of retrograde flow (RF) in lower extremity veins. METHODS: Eighty limbs in 40 healthy subjects and 60 limbs in 45 patients with chronic venous disease were examined with duplex scanning in the standing and supine positions. Each limb was assessed for reflux at 16 venous sites, including the common femoral, deep femoral, and proximal and distal femoral veins; proximal and distal popliteal veins; gastrocnemial vein; anterior and posterior tibial veins; peroneal vein; greater saphenous vein, at the saphenofemoral junction, thigh, upper calf, and lower calf; and lesser saphenous vein, at the saphenopopliteal junction and mid-calf. Perforator veins along the course of these veins were also assessed. In the healthy volunteers, 1553 vein segments were assessed, including 480 superficial vein segments, 800 deep vein segments, and 273 perforator vein segments; and in the patients, 1272 vein segments were assessed, including 360 superficial vein segments, 600 deep vein segments, and 312 perforator vein segments. Detection and measurement of reflux were performed at duplex scanning. Standard pneumatic cuff compression pressure was used to elicit reflux. Duration of RF and peak vein velocity were measured immediately after release of compression. RESULTS: Duration of RF in the superficial veins ranged from 0 to 2400 ms (mean, 210 ms), and was less than 500 ms in 96.7% of these veins. In the perforator veins, regardless of location, outward flow ranged from 0 to 760 ms (mean, 170 ms), and was less than 350 ms in 97% of these veins. In the deep veins, RF ranged from 0 to 2600 ms. Mean RF in the deep femoral veins and calf veins was 190 ms, and was less than 500 ms in 97.6% of these veins. In the femoropopliteal veins, mean RF was 390 ms, and ranged from 510 to 2600 ms in 21 of 400 segments; however, RF was less than 990 ms in 99% of these veins. Duration of RF was significantly longer in all three veins systems in patients (P <.0001 for all comparisons). With a cutoff value of more than 1000 ms rather than more than 500 ms, prevalence of abnormal RF in the femoropopliteal veins was significantly reduced, from 29% to 18% (P =.002). Thirty-seven vein segments (2.4%) had RF greater than 500 ms in the supine position, compared with less than 500 ms in 22 of these vein segments (59%) in the standing position. Of the 48 vein segments (3.1%) with RF greater than 500 ms in the standing position, RF was less than 500 ms in 6 of these vein segments (13%) in the supine position. Similar observations were noted in patient veins. There was no association between RF and peak vein velocity. Peak vein velocity had no significance in determining reflux. CONCLUSIONS: The cutoff value for reflux in the superficial and deep calf veins is greater than 500 ms. However, the reflux cutoff value for the femoropopliteal veins should be greater than 1000 ms. Outward flow in the perforating veins should be considered abnormal at greater than 350 ms. Reflux testing should be performed with the patient standing. PMID- 14560233 TI - Subfascial endoscopic perforating vein surgery as treatment for lateral perforating vein incompetence and venous ulceration. AB - INTRODUCTION: Endoscopic ligation of perforating veins (subfascial endoscopic perforating vein surgery [SEPS]) is valuable in treatment of venous ulcer on the medial aspect of the lower leg. Venous ulcerations on the lateral aspect of the lower leg are less common, but are of equal clinical importance. MATERIAL AND METHODS: Over 4 years we performed SEPS on the lateral aspect of the lower leg in 13 patients. Preoperative and postoperative duplex scanning of the perforating veins was performed in all patients. RESULTS: Mean follow-up was 53 months (range, 33-81 months). Complete healing of venous ulcer was achieved in six patients within 6 months. Ulceration persisted in six patients, and ulcer recurred in three patients after 17, 29, and 60 months, respectively. Postoperative duplex scans showed persistent insufficient perforating veins in about 25% of patients. CONCLUSION: There is a considerable difference between the results of SEPS on the medial side of the lower leg and the same procedure on the lateral side. SEPS on the lateral side of the lower leg, as performed in this small series, does not contribute to ulcer healing or recurrence. A considerable number of perforating veins are missed at surgery, resulting in persistent insufficient perforating veins postoperatively. The poor results of this study emphasize the need for adequate anatomic information to improve the surgical outcome. PMID- 14560234 TI - Prospective evaluation of chronic venous insufficiency based on foot venous pressure measurements and air plethysmography findings. AB - PURPOSE: The purpose of this study was to evaluate lower extremity venous function in patients with chronic venous insufficiency, with foot venous pressure (FVP) measurements and air plethysmography (APG). METHODS: Eighty-five limbs of 63 patients with a history of chronic venous insufficiency (CVI) from 1995 to 1999 were studied. FVP parameters studied included ambulatory venous pressure (AVP), percent decrease in FVP with manual calf compression (%drop), ratio of increase in FVP over 4 seconds after release of compression (4SR%), and time to 90% recovery of FVP were measured. APG parameters studied included functional venous volume, 90% refilling time (VFT90), venous filling index, ejection fraction, and residual volume fraction. RESULTS: Venous filling index and 90% refilling time were significantly decreased in limbs with stasis syndrome compared with the control group. AVP, %drop, and 4SR% also showed significantly decrease in limbs with stasis syndrome compared with those without it. AVP, %drop, and 4SR% were significantly different for the primary group compared with the secondary group, whereas no differences were found with regard to any APG parameter. CONCLUSIONS: APG enables prediction of the presence of CVI, whereas FVP measurements are more useful for evaluation of clinical severity of CVI. PMID- 14560235 TI - Overexpression of mutated IkappaBalpha inhibits vascular smooth muscle cell proliferation and intimal hyperplasia formation. AB - PURPOSE: Vascular injury and inflammation are associated with elaboration of a number of cytokines that signal through multiple pathways to act as smooth muscle cell (SMC) mitogens. Activation of the nuclear factor-kappa B (NF-kappaB) transcription factor is essential for SMC proliferation in vitro and is activated by vascular injury in vivo. Activation of NF-kappaB is controlled by several upstream regulators, including the inhibitors of kappa B (IkappaB). These proteins bind to and keep NF-kappaB inactivated. The purpose of this study was to determine whether adenoviral gene transfer of a mutated IkappaBalpha super repressor (AdIkappaBalphaSR) could inhibit development of intimal hyperplasia in vivo and to investigate how over-expression of this construct influences in vitro SMC proliferation and cell cycle regulatory proteins. METHODS: A rat carotid injury model was used to study prevention of intimal hyperplasia. Arteries were assayed 14 days after injury and infection with AdIkappaBalphaSR or adenoviral beta-galactosidase (AdLacZ). Untreated SMC or SMC infected with AdLacZ or AdIkappaBalphaSR were stimulated with 10% fetal bovine serum, interleukin-1beta, or tumor necrosis factor-alpha. Electrophoretic mobility shift assays were used to assay for NF-kappaB activation. Protein levels of IkappaBalpha and cyclin dependent kinase inhibitors p21(Cip1/Waf1) and p27(Kip1) were determined with Western blot analysis. Proliferation was measured with (3)H-thymidine incorporation assays. RESULTS: AdIkappaBalphaSR inhibited the development of intimal hyperplasia by 49% (P <.05). Infection with AdIkappaBalphaSR significantly suppressed in vitro SMC proliferation when stimulated with serum, interleukin 1, or tumor necrosis factor alpha, and did not result in cell death. Inhibition of proliferation was associated with increased p21(Cip1/Waf1) and p27(Kip1) protein levels. CONCLUSIONS: Gene transfer of IkappaBalpha super repressor inhibited development of intimal hyperplasia in vivo and SMC proliferation in vitro. The antiproliferative activity may be related to cell cycle arrest through upregulation of the cyclin-dependent kinase inhibitors p21 and p27. Overexpression of IkappaBalpha may be a future therapeutic option in treatment of vascular diseases. PMID- 14560237 TI - Abnormal mitochondrial respiration in skeletal muscle in patients with peripheral arterial disease. AB - OBJECTIVE: Discrete morphologic, enzymatic and functional changes in skeletal muscle mitochondria have been demonstrated in patients with peripheral arterial disease (PAD). We examined mitochondrial respiration in the gastrocnemius muscle of nine patients (10 legs) with advanced PAD and in nine control patients (nine legs) without evidence of PAD. METHODS: Mitochondrial respiratory rates were determined with a Clark electrode in an oxygraph cell containing saponin-skinned muscle bundles. Muscle samples were obtained from the anteromedial aspect of the gastrocnemius muscle, at a level 10 cm distal to the tibial tuberosity. Mitochondria respiratory rate, calculated as nanoatoms of oxygen consumed per minute per milligram of noncollagen protein, were measured at baseline (V(0)), after addition of substrates (malate and glutamate; (V(SUB)), after addition of adenosine diphosphate (ADP) (V(ADP)), and finally, after adenine nucleotide translocase inhibition with atractyloside (V(AT)). The acceptor control ratio, a sensitive indicator of overall mitochondrial function, was calculated as the ratio of the respiratory rate after the addition of ADP to the respiratory rate after adenine nucleotide translocase inhibition with atractyloside (V(ADP)/ V(AT)). RESULTS: Respiratory rate in muscle mitochondria from patients with PAD were not significantly different from control values at baseline (0.31 +/- 0.06 vs 0.55 +/- 0.12; P =.09), but V(sub) was significantly lower in patients with PAD compared with control subjects (0.43 +/- 0.07 vs 0.89 +/- 0.20; P <.05), as was V(ADP) (0.69 +/- 0.13 vs 1.24 +/- 0.20; P <.05). Respiratory rates after atractyloside inhibition in patients with PAD were no different from those in control patients (0.47 +/- 0.07 vs 0.45 +/- P =.08). Compared with control values, mitochondria from patients with PAD had a significantly lower acceptor control ratio (1.41 +/- 0.10 vs 2.90 +/- 0.20; P <.001). CONCLUSION: Mitochondrial respiratory activity is abnormal in lower extremity skeletal muscle in patients with PAD. When considered in concert with the ultrastructural and enzymatic abnormalities previously documented in mitochondria of chronically ischemic muscle, these data support the concept of defective mitochondrial function as a pathophysiologic component of PAD. PMID- 14560236 TI - Overexpression of endothelial nitric oxide synthase increases skeletal muscle blood flow and oxygenation in severe rat hind limb ischemia. AB - OBJECTIVE: Although nitric oxide (NO) has a critical role in angiogenesis, the therapeutic potential of NO synthase overexpression in severe ischemia remains undefined. We tested the hypothesis that overexpression of endothelial NO synthase (eNOS) would improve tissue perfusion in severe hind limb ischemia. METHODS: Severe hind limb ischemia was induced in 122 adult male Sprague-Dawley rats. Ten days after the induction of hind limb ischemia, vascular isolation and intraarterial delivery of an adenoviral vector encoding eNOS (AdeNOS), a control adenoviral vector (AdE1), or phosphate-buffered saline solution (PBS) was performed. Skeletal muscle blood flow, muscle oxygen tension, angiography, and immunohistochemistry for capillary counts were measured. RESULTS: Gene transfer of AdeNOS increased eNOS protein expression and enzyme activity. Two weeks after gene transfer, skeletal muscle blood flow was fourfold higher in eNOS-transduced than in AdE1-transduced or PBS treated rats and was similar to exercise-induced maximal flow in nonischemic muscle. eNOS overexpression increased muscle oxygen tension in a titer-dependent fashion. This increase persisted 1 month after transduction, even though eNOS enzyme activity had declined to normal levels. Angiography and capillary counts showed that eNOS overexpression increased the size and number of collateral arteries, but did not significantly increase the capillary-muscle fiber ratio. CONCLUSIONS: eNOS overexpression in an ischemic rat hind limb significantly increased skeletal muscle blood flow, muscle oxygen tension, and collateral arteries (arteriogenesis). Furthermore, eNOS overexpression did not result in capillary angiogenesis above control levels. These studies demonstrate the potential for eNOS overexpression as treatment for severe limb ischemia in human beings. PMID- 14560238 TI - Endovascular management of axillofemoral bypass graft stump syndrome. AB - OBJECTIVE: Upper extremity embolic complications of occluded axillofemoral bypass grafts are infrequent. However, traditional management of dissection of axillary anastomosis for removal of the stump can be challenging. We report two patients with critical upper extremity ischemia secondary to stump syndrome and its successful management with endovascular techniques. METHODS: One hundred fifty two patients underwent axillofemoral bypass grafting over 10 years from 1991 2001. Two patients from this series had acute ischemia involving the ipsilateral upper extremity of occluded axillofemoral bypass graft. Duplex ultrasound scans revealed occlusion of the axillofemoral bypass graft and acute occlusion of ipsilateral upper extremity arteries. Both patients underwent brachial artery exploration and embolectomy. Completion angiograms revealed persistent axillofemoral bypass graft stump as the source of embolus. The stump was obliterated with a 10-mm/40-mm Wallgraft introduced through the same arteriotomy made for brachial embolectomy. Transesophageal echocardiography and magnetic resonance angiography of the arch and great vessels were performed to exclude other sources of origin for the embolus. RESULTS: Both patients remained symptom free and with patent stent grafts, as seen on duplex scans at 3, 6, and 9 months of follow-up. CONCLUSIONS: Upper extremity embolism is a rare complication after occlusion of axillofemoral bypass grafts. The endovascular approach to obliterate the stump of occluded axillofemoral bypass grafts is minimally invasive and an effective alternative treatment of this rare condition. PMID- 14560239 TI - Endovascular stent-graft placement for nonaneurysmal infrarenal aortic rupture: a case report and review of the literature. AB - Penetrating atheromatous ulceration of the infrarenal aorta is a rare entity. There are few reported cases of this lesion, and most of the published data is in regards to the thoracic aorta. Spontaneous rupture of a nonaneurysmal noninfected atherosclerotic infrarenal aorta is a rare event. We report the eleventh case of this occurrence and present the first reported case of endovascular stent-graft placement in treating this entity. We review the literature regarding ulcerative disease of the aorta and specifically discuss the published data on spontaneous rupture of the nondilated, noninfected infrarenal aorta secondary to penetrating atheromatous ulceration. PMID- 14560240 TI - Delayed aortic aneurysm enlargement due to endotension after endovascular abdominal aortic aneurysm repair. AB - Abdominal aortic aneurysm (AAA) enlarges after successful endovascular repair, because of persistent blood flow within the aneurysm sac, or endoleak. In the absence of detectable endoleak, AAA may still expand, in part because of persistent pressurization within the excluded aneurysm, or endotension. We report three patients who underwent successful endovascular AAA repair in whom postoperative surveillance showed aneurysm regression, yet delayed AAA enlargement without demonstrable endoleak developed in all three patients. Endotension was confirmed in all three patients at elective open conversion. Our study underscores the significance of endotension as a mechanism of delayed aneurysm enlargement after successful endovascular AAA repair. PMID- 14560241 TI - Fabric tears as a new cause of type III endoleak with Ancure endograft. AB - PURPOSE: We present two case reports of type IIIb endoleak. One was due to fabric erosion caused by placement of a stent (Wallstent; W. L. Gore & Associates, Flagstaff, Ariz) after endovascular aneurysm repair; the other arose spontaneously. In both cases, an Ancure endograft (Guidant/EVT, Menlo Park, Calif) was placed. CASE REPORTS: In case 1, a large endoleak developed 36 months after uncomplicated endovascular treatment of an abdominal aortic aneurysm. In case 2, endoleak developed 30 months after a complicated procedure. In both cases, two Wallstents were used to treat type I endoleak and limb kinking in the first postoperative months. One type III endoleak was within the endograft at the level of the stents. CONCLUSION: To our knowledge, these are the first type III endoleaks reported in association with Ancure endografts. Placement of Wallstents inside endografts is of concern, and another indication for close monitoring during follow-up. PMID- 14560242 TI - Mycotic aneurysm of the infrarenal abdominal aorta infected by Clostridium septicum: a case report of surgical management and review of the literature. AB - We report a surgical case of mycotic aneurysm of the infrarenal abdominal aorta infected by Clostridium septicum. The patient was first treated with an in situ prosthetic graft replacement. When the infection recurred 5 weeks after the aortic surgery, the patient was successfully treated by transposition of rectus abdominis muscle flap around the graft. Only 19 cases of mycotic aneurysm or aortic dissection caused by Clostridium septicum have been reported. Ten of 12 patients who underwent vascular surgery survived, whereas all 7 patients who did not undergo surgery died. Surgical treatment should be undertaken since the surgical results seem satisfactory. PMID- 14560243 TI - Autologous deep vein reconstruction of infected thoracoabdominal aortic patch graft. AB - Graft infection remains a serious complication of prosthetic aortic repair. Infection of thoracoabdominal aortic prosthetic grafts, in particular, is a significant clinical challenge and is associated with high mortality. We report successful in situ reconstruction of an infected thoracoabdominal aortic prosthetic patch graft with autogenous superficial femoral vein. To our knowledge, this is the first such case described in the North American and English language surgical literature. At 24-month follow-up the patient remains well, with no evidence of sepsis or graft complication at clinical and radiologic assessment. PMID- 14560244 TI - Branched endovascular repair of aortic arch aneurysm with a modular stent-graft system. PMID- 14560246 TI - Modular branched stent graft for endovascular repair of aortic arch aneurysm and dissection. AB - PURPOSE: We describe a modular stent graft for use in endovascular repair of aneurysms of the aortic arch. METHOD: Carotid-carotid and left carotid-subclavian bypass grafts are created surgically. Two large, fully stented grafts are inserted endoluminally. The proximal component is bifurcated, with a wide proximal trunk and two distal limbs, one long and narrow, the other short and wide. This component is inserted through the carotid artery and deployed with the trunk and short wide limb in the ascending thoracic aorta; the long narrow limb opens into the innominate artery. After delivery system removal and carotid artery repair, a distal component is inserted through a femoral approach to bridge the gap between the short, wide distal limb of the proximal component and the nondilated descending thoracic aorta. The result is a branched stent graft, implanted proximally into the ascending aorta and distally into the innominate artery and descending thoracic aorta. CONCLUSION: The system has been used successfully to treat a large wide-necked pseudoaneurysm of the aortic arch. PMID- 14560245 TI - Endovascular assisted in situ bypass grafting: a simplified technique for saphenous vein side branch occlusion. AB - The in situ bypass procedure for lower extremity limb salvage requires a long continuous incision or multiple interrupted incisions over the greater saphenous vein to ligate the saphenous vein side branches. This can result in wound complications that frequently prolong hospital stay and threaten the graft. In an effort to reduce the incidence of wound complications, alternate methods of occluding the vein side branches have been used. One method is to deliver coils under angioscopic vision into the saphenous vein side branches. This report details a simplified technique that uses widely available catheter-based equipment to perform saphenous vein side branch occlusion under fluoroscopic guidance. PMID- 14560247 TI - Autogenous arterial radial-basilic forearm looped transposition. AB - We describe the use of radial artery as a conduit for arteriovenous hemodialysis access in the forearm. The advantages of this type of fistula are use of autogenous material and its localization because forearm fistula is easily cannulated and is more convenient for the patient. This fistula may be an alternative solution in selected hemodialysis patients for the difficult access in whom superficial arm veins are exhausted and lower extremity access is unsuitable. PMID- 14560248 TI - What to tell patients harmed by other physicians. PMID- 14560262 TI - Who provides orthodontic treatment? PMID- 14560263 TI - Principles of retention in orthodontia. 1920. PMID- 14560264 TI - Rapid palatal expansion in the absence of crossbites: added value? PMID- 14560265 TI - Distribution of orthodontic services and fees in an insured population in Washington. AB - Previous attempts to quantify the amount and type of orthodontic therapy provided by nonorthodontists in the United States have relied on survey data. Although there are advantages to surveys, such as control over survey recipients and inclusion of specific questions, they also have limitations, such as low response rates, response bias, and recall bias. This study used insurance claims data from a large dental benefits provider in Washington to assess the distribution of orthodontic services and fees among various dental providers. All orthodontic claims allowed by Washington Dental Service in 2001 were retrieved, along with treatment codes, fees, and demographic information for both patients and providers. A total of 102,984 orthodontic claims were included in the study. General dentists submitted 7.0% of these claims, orthodontists submitted 90.9%, and pedodontists submitted 1.9%. Orthodontists submitted higher average fees for space maintainers, first payments, and records. The percentage of orthodontic treatment preformed by general dentists and pedodontists in this claims-based study was substantially less than what has been previously reported in survey based studies. Additionally, a smaller percentage of general dentists and pedodontists in this study performed comprehensive treatment, compared with previous studies. This study illustrates the value of insurance claims data to assess the provision of orthodontic care. PMID- 14560266 TI - Factors associated with the stability of titanium screws placed in the posterior region for orthodontic anchorage. AB - Recently, implant anchors such as titanium screws have been used for absolute anchorage during edgewise treatment. However, there have been few human studies reporting on the stability of implant anchors placed in the posterior region. The purpose of this study was to examine the success rates and to find the factors associated with the stability of titanium screws placed into the buccal alveolar bone of the posterior region. Fifty-one patients with malocclusions, 134 titanium screws of 3 types, and 17 miniplates were retrospectively examined in relation to clinical characteristics. The 1-year success rate of screws with 1.0-mm diameter was significantly less than that of other screws with 1.5-mm or 2.3-mm diameter or than that of miniplates. Flap surgery was associated with the patient's discomfort. A high mandibular plane angle and inflammation of peri-implant tissue after implantation were risk factors for mobility of screws. However, we could not detect a significant association between the success rate and the following variables: screw length, kind of placement surgery, immediate loading, location of implantation, age, gender, crowding of teeth, anteroposterior jaw base relationship, controlled periodontitis, and temporomandibular disorder symptoms. We concluded that the diameter of a screw of 1.0 mm or less, inflammation of the peri-implant tissue, and a high mandibular plane angle (ie, thin cortical bone), were associated with the mobility (ie, failure) of the titanium screw placed into the buccal alveolar bone of the posterior region for orthodontic anchorage. PMID- 14560267 TI - Mechanical and biological comparison of latex and silicone rubber bands. AB - Latex rubber bands are routinely used to supply orthodontic force. However, because the incidence of allergic reactions to latex is rising, the use of nonlatex alternatives is increasing, and assessing the mechanical properties of the replacement products is becoming more important. The purposes of this study were to compare the mechanical properties of latex and silicone orthodontic rubber bands through static testing under dry and wet conditions, and to compare their biologic (cytotoxic) properties. Three brands of latex and 1 brand of silicone rubber bands were tested. When extended to 300% of the lumen diameter, the silicone group had an initial force equal to 83% of the product specifications; this was the lowest of the 4 groups. All 4 brands showed notable amounts of force degradation at the 300% extension when subjected to saliva immersion; this approximated a 30% force decay over 2 days. The latex bands all followed a similar pattern of force degradation, whereas the silicone bands showed a greater increase in force decay as the extension length increased. The silicone bands were less cytotoxic than 2 of the 3 types of latex. Although the silicone bands showed the least discrepancy of force degradation between air and saliva conditions, the amount of the force decay was the greatest. Therefore, great improvements in the physical properties of the silicone band are required before they can be considered an acceptable replacement for latex. PMID- 14560268 TI - Temperature-modulated DSC provides new insight about nickel-titanium wire transformations. AB - Differential scanning calorimetry (DSC) is a well-known method for investigating phase transformations in nickel-titanium orthodontic wires; the microstructural phases and phase transformations in these wires have central importance for their clinical performance. The purpose of this study was to use the more recently developed technique of temperature-modulated DSC (TMDSC) to gain insight into transformations in 3 nickel-titanium orthodontic wires: Neo Sentalloy (GAC International, Islandia, NY), 35 degrees C Copper Ni-Ti (Ormco, Glendora, Calif) and Nitinol SE (3M Unitek, Monrovia, Calif). In the oral environment, the first 2 superelastic wires have shape memory, and the third wire has superelastic behavior but not shape memory. All wires had cross-section dimensions of 0.016 x 0.022 in. Archwires in the as-received condition and after bending 135 degrees were cut into 5 or 6 segments for test specimens. TMDSC analyses (Model 2910 DSC, TA Instruments, Wilmington, Del) were conducted between -125 degrees C and 100 degrees C, using a linear heating and cooling rate of 2 degrees C per min, an oscillation amplitude of 0.318 degrees C with a period of 60 seconds, and helium as the purge gas. For all 3 wire alloys, strong low-temperature martensitic transformations, resolved on the nonreversing heat-flow curves, were not present on the reversing heat-flow curves, and bending appeared to increase the enthalpy change for these peaks in some cases. For Neo Sentalloy, TMDSC showed that transformation between martensitic and austenitic nickel-titanium, suggested as occurring directly in the forward and reverse directions by conventional DSC, was instead a 2-step process involving the R-phase. Two-step transformations in the forward and reverse directions were also found for 35 degrees C Copper Ni-Ti and Nitinol SE. The TMDSC results show that structural transformations in these wires are complex. Some possible clinical implications of these observations are discussed. PMID- 14560269 TI - Evaluation of friction of stainless steel and esthetic self-ligating brackets in various bracket-archwire combinations. AB - This study measured and compared the level of frictional resistance generated between stainless steel self-ligating brackets (Damon SL II, SDS Ormco, Glendora, Calif), polycarbonate self-ligating brackets (Oyster, Gestenco International, Gothenburg, Sweden), and conventional stainless steel brackets (Victory Series, 3M Unitek, Monrovia, Calif), and 3 different orthodontic wire alloys: stainless steel (Stainless Steel, SDS Ormco), nickel-titanium (Ni-Ti, SDS Ormco), and beta titanium (TMA, SDS Ormco). All brackets had a.022-in slot, whereas the orthodontic wire alloys were tested in 3 different sections:.016,.017 x.025, and.019 x 0.025 in. Each of the 27 bracket and archwire combinations was tested 10 times, and each test was performed with a new bracket-wire sample. Both static and kinetic friction were measured on a custom-designed apparatus. All data were statistically analyzed (Kruskal-Wallis and Mann Whitney U tests). Stainless steel self-ligating brackets generated significantly lower static and kinetic frictional forces than both conventional stainless steel and polycarbonate self ligating brackets, which showed no significant differences between them. Beta titanium archwires had higher frictional resistances than stainless steel and nickel-titanium archwires. No significant differences were found between stainless steel and nickel-titanium archwires. All brackets showed higher static and kinetic frictional forces as the wire size increased. PMID- 14560271 TI - Shear bond strength of orthodontic brackets bonded with a modified 1-step etchant and-primer technique. AB - When bonding orthodontic brackets to enamel, most orthodontists use a conventional technique of etching tooth enamel with phosphoric acid, placing a hydrophilic primer on the etched, rinsed, and dried surface, then bonding the bracket with an adhesive resin composite. New systems simultaneously etch and prime the tooth surface in preparation for bonding. The purpose of this study was to compare the shear bond strength of orthodontic brackets bonded to enamel with a conventional, multistep adhesive system and a self-etching primer adhesive system. In addition, a third group was included in which the air dispersion step in the self-etching primer system was omitted. Brackets were bonded to 108 extracted human molars according to 1 of 3 experimental protocols--group 1: conventional multistep adhesive (n = 36); group 2: self-etching primer system (n = 36); group 3: self-etching primer system without air dispersion (n = 36). Specimens were loaded to failure in a universal testing machine (Instron, Canton, Mass). Mean shear bond strengths in megapascals (standard deviation) were 11.3 (2.2), 11.9 (3.2), and 8.2 (2.8) for groups 1, 2, and 3, respectively. Data were subjected to 1-way analysis of variance at alpha =.05. The mean shear bond strength of the self-etching primer group in which the air dispersion step was omitted was significantly less than in the other 2 groups (P <.001). However, there was no difference in mean shear bond strength between the conventional, multistep adhesive system and the self-etching primer system when the primer was dispersed correctly (P =.34). PMID- 14560270 TI - Evaluation of friction of conventional and metal-insert ceramic brackets in various bracket-archwire combinations. AB - The purpose of the study was to measure and compare the level of frictional resistance generated between conventional ceramic brackets (Transcend Series 6000, 3M Unitek, Monrovia, Calif), ceramic brackets with stainless steel slot (Clarity, 3M Unitek), conventional stainless steel brackets (Victory Series, 3M Unitek), and 3 different orthodontic wire alloys: stainless steel (stainless steel, SDS Ormco, Glendora, Calif), nickel-titanium (Ni-Ti, SDS Ormco), and beta titanium (TMA, SDS Ormco). All brackets had a 0.022-in slot, and orthodontic wire alloys were tested in 3 different sections: 0.016 in, 0.017 x 0.025 in, and 0.019 x 0.025 in. Each of the 27 bracket-archwire combinations was tested 10 times, and each test was performed with a new bracket-wire sample. Static and kinetic friction were measured on a specially designed apparatus. All data were statistically analyzed (analysis of variance and Scheffe for the bracket effect, Kruskal-Wallis and Mann Whitney for the alloy and section effects). Metal-insert ceramic brackets generated significantly lower frictional forces than did conventional ceramic brackets, but higher values than stainless steel brackets, in agreement with the findings of the few previous reports. Beta-titanium archwires had higher frictional resistances than did stainless steel and nickel titanium archwires. No significant differences were found between stainless steel and nickel-titanium archwires. All the brackets showed higher static and kinetic frictional forces as the wire size increased. Metal-insert ceramic brackets are not only visually pleasing, but also a valuable alternative to conventional stainless steel brackets in patients with esthetic demands. PMID- 14560272 TI - Effect of saliva on shear bond strength of an orthodontic adhesive used with moisture-insensitive and self-etching primers. AB - The purpose of this study was to investigate the effect of saliva contamination on the shear bond strength of an orthodontic adhesive used with Transbond Moisture-Insensitive Primer (MIP, 3M Unitek, Monrovia, Calif) and Transbond Plus Self-Etching Primer (SEP, 3M Unitek). Hydrophobic Transbond XT primer (XT, 3M Unitek) was used as a control. A total of 162 extracted premolars were collected and divided equally into 9 groups of 18 teeth each, and brackets were bonded with Transbond XT adhesive (3M Unitek) under different experimental conditions: (1) control: etch/dry/XT, (2) etch/dry/MIP, (3) etch/dry/MIP/wet (saliva)/MIP, (4) etch/wet/MIP, (5) etch/wet/MIP/wet/MIP, (6) dry/SEP, (7) dry/SEP/wet/SEP, (8) wet/SEP, and (9) wet/SEP/wet/SEP. Shear bond strength of each sample was examined with a testing machine. The results showed that the control group had the highest mean shear bond strength (group 1, 21.3 +/- 6.8 MPa), followed by the MIP group in a dry field (group 2, 20.7 +/- 5.0 MPa). No significant difference was found between groups 1 and 2. Groups 3 through 9 had similar mean strengths, ranging from 12.7 to 15.0 MPa (P >.05), which were significantly lower than in groups 1 and 2 (P <.05). There was no significant difference in bond-failure site among the 9 groups. It was concluded that (1) Transbond XT adhesive with Transbond XT primer and MIP in a dry field yields similar bond strengths, which are greater than all other groups, (2) saliva contamination significantly lowers the bond strength of Transbond MIP, (3) saliva has no effect on the bond strength of Transbond SEP, (4) Transbond XT adhesive with Transbond MIP and SEP might have clinically acceptable bond strengths in either dry or wet fields. PMID- 14560273 TI - Use of a self-etching primer in combination with a resin-modified glass ionomer: effect of water and saliva contamination on shear bond strength. AB - The purpose of this study was to evaluate the effects of 3 different enamel conditioners (10% polyacrylic acid, 37% phosphoric acid, and self-etching primer) on the shear bond strength and site of bond failure of a resin-modified glass ionomer (Fuji Ortho LC, GC Europe, Leuven, Belgium) bonded onto dry, water moistened, and saliva-moistened enamel. One hundred eighty bovine permanent mandibular incisors were randomly divided into 12 groups; each group consisted of 15 specimens. Three different enamel surface conditions were studied: dry, soaked with water, soaked with saliva. One hundred eighty stainless steel brackets were bonded with the resin-modified glass ionomer. After bonding, all samples were stored in distilled water for 24 hours and then tested in a shear mode on a testing machine. After self-etching primer application, Fuji Ortho LC produced the highest shear bond strengths under all the different enamel surface conditions; these values were significantly higher than those achieved in the remaining groups, except when Fuji Ortho LC was used in combination with 37% phosphoric acid on dry enamel. Fuji Ortho LC bonded without enamel conditioning produced the lowest shear bond strengths. The bond strength of the groups conditioned with 10% polyacrylic acid was significantly lower than that of the groups etched with 37% phosphoric acid, except when both conditioners were used on enamel soaked with water. PMID- 14560274 TI - Effect of ultrasound on rabbit mandibular incisor formation and eruption after mandibular osteodistraction. AB - Therapeutic ultrasound is known to enhance bone-fracture healing. The purpose of this article was to study the effect of therapeutic ultrasound on mandibular incisor development and eruption in 15 skeletally mature male New Zealand white rabbits undergoing mandibular osteodistraction. The surgical cuts were made anterior to the molars and consequently transected the mandibular incisors. The animals were divided into 3 groups of 5 rabbits each. Group I received ultrasound on both sides of the mandible every other day. Group II received ultrasound on the left side of the mandible every day. Group III did not receive ultrasound. Ultrasound was given for 20 minutes per application. Mandibular incisor eruption was measured directly on the rabbits and on lateral head radiographs. The animals were killed 4 weeks after surgery, and dissected mandibular specimens that contained the incisor segments were evaluated histologically. The lengths of the tooth segments increased in the ultrasound-treated animals, compared with the controls (group III). However, the rabbits in group II had greater increases in length of the mandibular incisor segments than those in group I. Histologic examination showed that the newly formed tissues at the distraction site and at the cut ends of the incisor segments consisted of osteodentin-like tissue and cementum. Thus, therapeutic ultrasound combined with distraction enhanced mandibular incisor growth and eruption. PMID- 14560275 TI - Class III malocclusion with severe facial asymmetry, unilateral posterior crossbite, and temporomandibular disorders. AB - A 22-year-old woman had a Class III malocclusion with severe facial asymmetry, unilateral posterior crossbite, and temporomandibular disorders. A clicking sound was noted in the temporomandibular joint on the posterior crossbite side during jaw opening, and she complained of pain in the masticatory muscles on both sides. The articular disc on the crossbite side was displaced anteriorly without reduction. The patient was treated orthodontically with edgewise appliances and surgically with LeFort I and intraoral vertical ramus osteotomies. The result of the combined surgical-orthodontic treatment was facial symmetry and optimal occlusion. The displaced articular disc moved into a normal position, and most of the temporomandibular disorder symptoms improved. At the 2.5-year follow-up, the temporomandibular joint conditions had been maintained. PMID- 14560276 TI - Early and unusual incisor resorption due to impacted maxillary canines. AB - A very early and severe case of maxillary incisor resorption caused by impacted canines is reported. An estimated 50,000 cases of ectopic eruption and impaction of maxillary canines occur each year in the United States. Although incisor resorption due to ectopically positioned permanent maxillary canines can be swift, silent, and devastating, an effective protocol has been developed for early detection and management of this condition. Palpation and, if indicated, radiographic evaluation are combined with primary canine removal in selected cases. These strategies--particularly when used early--can prevent the vast majority of palatally impacted maxillary canines and the potentially devastating resorption of adjacent incisors. PMID- 14560278 TI - Litigation, legislation, and ethics. You can't say that. PMID- 14560277 TI - Solid-state temporomandibular joint imaging: accuracy in detecting osseous changes of degenerative joint disease and determining condylar spatial relations. AB - The purpose of this study was to evaluate the off-label use of an intraoral charge-coupled device (CCD) for extraoral transcranial radiography of the temporomandibular joint. Corrected linear tomograms and transcranial images made with conventional screen-film combinations and a CCD detector were compared with sectioned cadaver specimens. Radiation dosage, qualitative assessment of condylar degenerative features, and condylar position within the glenoid fossa of the 3 modalities were assessed and compared. The CCD method required special adjustments to achieve adequate quality, and it involved greater exposure than the other methods. This use of this intraoral system for extraoral imaging cannot now be recommended, but future refinements might make it more viable. PMID- 14560279 TI - Advanced practice nursing: issues unresolved and requiring feedback. PMID- 14560280 TI - Pressure ulcers and lateral rotation beds: a case study. PMID- 14560281 TI - Obtaining informed consent for participation from cognitively impaired adults. PMID- 14560282 TI - Does vitamin C supplementation promote pressure ulcer healing? PMID- 14560284 TI - Community health nursing, wound care, and...ethics? AB - Because of changing demographics and other factors, patients receiving care for wounds, ostomies, or incontinence are being referred in increasing numbers to community health nursing organizations for initial or continued care. As home based wound care becomes big business, little discussion is being focused on the moral and ethical issues likely to arise in the high-tech home setting. Progressively more complex and expensive home care relies on family members to take on complicated care regimens in the face of decreasing numbers of allowable skilled nursing home visits. A framework and a principle-based theory for reflection on the character and content of moral and ethical conflicts are provided to encourage informed and competent care of patients in the home. Common moral and ethical conflicts for WOC nurses in the United States are presented. These conflicts include issues of wound care supply procurement; use of documentation to maximize care or profit; problems of quality, care consistency, and caregiver consent; and dilemmas of tiered health care options. The advantages of a framework to address ethical conflicts are discussed. PMID- 14560283 TI - Clinical trial of a prevention and treatment protocol for skin breakdown in two nursing homes. AB - OBJECTIVE: Our objective was to assess the effectiveness of skin care protocols, including a body wash and skin protectant, on skin breakdown in 2 nursing homes. DESIGN: This was a quasi-experimental pretest/posttest design study. Setting and subjects Adult residents (n = 136) of 2 skilled nursing homes consented to participate in this study. Seventy percent were women; the sample average age of 82 years. INSTRUMENTS: A researcher-designed data recording form documented resident demographics, incidence and type of skin breakdown or pressure ulcer, presence of urinary or fecal incontinence, and assessment of the effectiveness of body wash and skin protectant. METHODS: Baseline data on prevalence of pressure ulcers and skin protocol were collected weekly for a 3-month period followed by a week-long educational program by the researchers about skin care and the body wash and skin protectant. During the 3-month trial with the body wash and skin protectant incorporated into routine care, research assistants recorded resident data weekly and researchers again assessed prevalence and incidence of pressure ulcers and skin breakdown weekly. RESULTS: Incorporation of a body wash and skin protectant into a skin care prevention and early intervention protocol in 2 nursing homes documented a decrease in skin breakdowns from 68 pre-intervention to 40 postintervention; the decrease in agency B was statistically significant. There was a statistically significant decrease in stage I and II pressure ulcer incidence overall (pre-intervention = 19.9%, postintervention = 8.1%). Nurses evaluated the body wash and skin protectant as effective for 98% of the time used. CONCLUSION: Implementation of a protocol for skin care along with staff education, including the prophylactic use of a body wash and skin protectant, reduced the incidence of skin breakdown, including pressure ulcers and perineal dermatitis, in 2 long-term care facilities. PMID- 14560285 TI - Replicating a survey of pressure ulcer content in nursing textbooks. AB - Nurses enter the profession with knowledge about pressure ulcer prevention and care and acquired from their basic educational programs. A prior review of nursing textbooks regarding pressure ulcer content published almost a decade ago revealed that incomplete, inaccurate, and at times limited information (as few as 200 lines of text) was contained in 10 commonly used textbooks. A review of the updated editions of the textbooks revealed a wide variation in the number of lines of text devoted to pressure ulcers, depth and quality of information, and the number of illustrations and tables. New findings were the inclusion of cultural concepts and research studies. These findings suggest an improvement in the amount and quality of pressure ulcer information in the majority of nursing textbooks surveyed. Recommendations for future textbook chapters on pressure ulcers are provided. PMID- 14560286 TI - Case study of a patient with a distal jejunostomy. AB - A young man with a short but complicated medical history was seen in our enterostomal therapy department in a large acute-care facility. The patient had a distal jejunostomy with an extremely high and problematic output. Cases such as his force WOC nurses to review basic anatomy, physiology, and psychosocial development to provide holistic care in a specialized practice. Such cases can justify the existence of WOC nurses and prove they are a valuable asset in the health care system. PMID- 14560287 TI - Analysis of interactive continence health information on the Web. AB - OBJECTIVE: The purpose of this study was to describe interactive information about continence health promotion for women that is available on Web sites identified by popular search engines or health/medical search engines or that is available on community-based or select health organizations' Web sites. METHODS: We used a variety of search services to search the Web for sites that disseminated information about continence health for women, for example, urinary incontinence, urine loss, and bladder problems. Two reviewers initially evaluated sites to determine if an interactive feature that provided individualized feedback was present. Web sites that had an interactive-with-feedback feature were further examined to delineate the purpose of the site and other descriptive characteristics. We evaluated each site according to content pertaining to continence health information. RESULTS: Most major search engines located more than 100,000 Web sites in the initial search; narrowing the search terms with the words "and women" decreased the number of sites. Only 13 sites with interactive with-feedback features were located for further evaluation. E-mail was the most common interactive feature; none of the sites contained comprehensive information on continence health promotion. CONCLUSIONS: Web sites exist and are informative in terms of continence health promotion, but they lack an emphasis on self-care and sufficient information to promote it. The functionality of computer technology and the Web has not been maximized for the purpose of delivering continence health information. PMID- 14560288 TI - Shouldn't we give seniors the opportunity for a less traumatic death? PMID- 14560289 TI - A couple's last journey. PMID- 14560291 TI - Sleep in the older adult: implications for nurses (CE). AB - Normal aging is accompanied by sleep pattern changes that may result in daytime sleepiness and affect the quality of life in older adults. These changes also can signal more serious sleep problems, which become more prevalent with aging. More than half of adults older than 65 living at home and about two-thirds of those in long-term care facilities have some difficulties with sleep. It is important for nurses to understand the normal changes in sleep that accompany aging and be able to differentiate them from the symptoms of serious sleep disorders to ensure holistic care for older adults. PMID- 14560290 TI - Bacteriuria detection with a urine dipstick applied to incontinence pads of nursing home residents. AB - The purpose of this study was to evaluate a new method of initial on-site evaluation of bacteriuria in an incontinent nursing home population. Nitrite and a composite of nitrite and/or leukocyte esterase results of the new method were compared with clean-catch urine culture results of each participant to determine sensitivity, specificity, positive and negative predictive values, and efficiency. These findings were compared with results of the traditional dipstick urinalysis for bacteriuria assessment. The new method was as effective as the traditional method in assessing both the presence and absence of bacteriuria. Results of this study indicate the new dipstick/pad method may assist in the assessment of bacteriuria in incontinent nursing home residents. PMID- 14560292 TI - Is there a nurse in the house? The role of nurses in assisted living: past, present, and future. AB - The large increase in the number of older adults in the United States has mandated more supportive housing options. Assisted living facilities (ALFs) were developed as a necessary, desirable, and available residential option for older adults. The lower cost of ALFs compared with skilled nursing facilities and the emphasis on greater autonomy within these facilities are appealing to older consumers and their families. However, although many ALF residents have chronic health care needs, the roles and availability of nurses in these facilities vary greatly. This article discusses the history and creation of ALFs, the current role of nurses within these facilities, and the many factors influencing nursing in the assisted living industry. The role of nurse practitioners in these facilities is discussed. Finally, a vision for the future of nursing in ALFs is presented. PMID- 14560293 TI - Conducting and interpreting interface pressure evaluations of clinical support surfaces. AB - Interface pressure measurement is a common method to compare various mattresses, operating room beds, or other clinical support surfaces used for patients. The colorful graphs create a good visual display of information. Unfortunately, no standard method exists for performing the measurements or displaying the information. Because the graph is such a good visual representation of the data, many clinicians rely on the look of the graph. However, altering its appearance by simple manipulations of the display characteristics is very easy. This manipulation can lead to inaccurate conclusions about the product and consequent purchase of products that are inappropriate, less effective, or more expensive than others. This article provides basic information about interface pressure measurement and interpretation so that clinicians can make informed decisions about various clinical surfaces. PMID- 14560294 TI - The clinical record: recognizing its value in litigation. AB - Documenting completely and accurately is standard nursing practice, yet many nurses do not seem to understand that proper documentation is crucial to the communication and critical decision-making necessary to meet patients' needs. In fact, failure to document can have lethal consequences. Documentation of intake and output (I & O) is used here as an example of the evidence attorneys and nurse experts look for in the clinical record of elders. From exhibits of I & O records, juries can determine if nurses collected sufficient data for appropriate decision-making to protect the patient from harm. Readers are invited to test themselves to determine if their own I & O documentation meets acceptable standards of nursing practice. If readers find areas for improvement, self-study or group in-services can be held to improve the documentation and use of these data. PMID- 14560296 TI - Irene Mortenson Burnside: her light shines on. PMID- 14560299 TI - Inappropriate prescribing practices and implications for nurses. PMID- 14560298 TI - The effect of bright light on sleep and behavior in dementia: an analytic review. AB - The purpose of this study was to examine bright light as a means of intervention in sleep-wakefulness and behavior disorder among patients with dementia through a review of the relevant literature. Fourteen articles were reviewed in depth for study design, intervention protocols, concurrent control groups, subject characteristics, and outcome measures of behavior and sleep. Previous studies found that behavior disorders improved, although not all the studies reviewed reported a quantitative measure of improvement. The effects on sleep disorders were controversial. As a result of limitations in past studies, the effect of bright light on dementia patients is inconclusive. The current review has revealed areas of weakness in previous research that might profitably be explored in the future. PMID- 14560301 TI - Assessing home safety in homebound older adults. PMID- 14560302 TI - Impact of the eating environment. PMID- 14560303 TI - Extent of mitochondrial DNA sequence variation in Atlantic cod from the Faroe Islands: a resolution of gene genealogy. AB - Variation in a 250 base pair (bp) fragment of the mitochondrial cytochrome b (cyt b) has been used extensively for population studies in Atlantic cod Gadus morhua. To study the shape of the gene genealogy and the nature of the polymorphism, sequences of another region of the cyt b gene and the TP intergenic spacer were added, making a total of 566 bp from 74 cod from the Faroe Islands. A total of 44 segregating sites defined 41 haplotypes, many at frequencies greater than 5%. Haplotype diversity was 0.97 and nucleotide diversity 0.73% per base. A topology referred to as a constellation gene genealogy was observed with four major haplotypes at high frequencies, from each of which a number of rare variants were derived. A young relative age of the haplotypes was gauged from the structure of the genealogy. The variation was mostly at synonymous sites within the coding region and thus likely to be neutral or under weak purifying selection. By comparative analysis this also applies to the TP spacer. Applying the locus to study population variation in the Faroe Islands by AMOVA revealed that the overall areas and localities within areas accounted for none of the variation, and all the variation was due to differences among individuals. PMID- 14560305 TI - Biodiversity: a climate for colonization. PMID- 14560307 TI - MECP2 analysis in mentally retarded patients: implications for routine DNA diagnostics. AB - Rett syndrome (RTT) is one of the most common neurodevelopmental disorders in females. The disease is caused by mutations in the methyl-CpG-binding protein 2 gene (MECP2), and various mutations have been reported. The phenotypic spectrum in both female and male patients is diverse, ranging from very mild to congenital encephalopathy and prenatal lethality. In this study, the question was addressed as to whether implementation of systematic screening of MECP2 in patients with an unexplained mental retardation in DNA diagnostics would be reasonable, and the spectrum of phenotypes resulting from mutations in this gene was further explored. Mutational analysis of MECP2 was performed in mentally retarded female patients who were negative for FMR1 CGG repeat expansion, in male and female patients with clinical features suggestive of either Angelman or Prader-Willi syndrome without methylation defects on chromosome 15q11-q13. In the cohort of females negative for the molecular Fragile-X studies (N=92), one nonsense mutation (p.Q406X) was found. In the cohort of Angelman-negative patients (N=63), two missense mutations (p.R133C in a female patient and a mosaic p.T158M in a male patient) were found, which have been reported many times in patients with classical RTT syndrome. In the Prader-Willi-negative group (N=98), no pathogenic mutations were found. The results support testing of patients with features suggestive of Angelman syndrome, but without methylation defects on chromosome 15q11-q13 for mutations in MECP2. In the remaining patients with unexplained mental retardation, additional clinical features should determine whether analysis of MECP2 is indicated. PMID- 14560308 TI - Phenotypic and molecular characterisation of the Aarskog-Scott syndrome: a survey of the clinical variability in light of FGD1 mutation analysis in 46 patients. AB - Faciogenital dysplasia or Aarskog-Scott syndrome (AAS) is a genetically heterogeneous developmental disorder. The X-linked form of AAS has been ascribed to mutations in the FGD1 gene. However, although AAS may be considered as a relatively frequent clinical diagnosis, mutations have been established in few patients. Genetic heterogeneity and the clinical overlap with a number of other syndromes might explain this discrepancy. In this study, we have conducted a single-strand conformation polymorphism (SSCP) analysis of the entire coding region of FGD1 in 46 AAS patients and identified eight novel mutations, including one insertion, four deletions and three missense mutations (19.56% detection rate). One mutation (528insC) was found in two independent families. The mutations are scattered all along the coding sequence. Phenotypically, all affected males present with the characteristic AAS phenotype. FGD1 mutations were not associated with severe mental retardation. However, neuropsychiatric disorders, mainly behavioural and learning problems in childhood, were observed in five out of 12 mutated individuals. The current study provides further evidence that mutations of FGD1 may cause AAS and expands the spectrum of disease causing mutations. The importance of considering the neuropsychological phenotype of AAS patients is discussed. PMID- 14560309 TI - Genome-wide scanning for linkage in Finnish breast cancer families. AB - Only a proportion of breast cancer families has germline mutations in the BRCA1 or BRCA2 genes, suggesting the presence of additional susceptibility genes. Finding such genes by linkage analysis has turned out to be difficult due to the genetic heterogeneity of the disease, phenocopies and incomplete penetrance of the mutations. Isolated populations may be helpful in reducing the level of genetic heterogeneity and in providing useful starting points for further genetic analyses. Here, we report results from a genome-wide linkage analysis of 14 high risk breast cancer families from Finland. These families tested negative for BRCA1 and BRCA2 germline mutations and showed no linkage to the 13q21 region, recently proposed as an additional susceptibility locus. Suggestive linkage was seen at marker D2S364 (2q32) with a parametric two-point LOD score of 1.61 (theta=0), and an LOD score of 2.49 in nonparametric analyses. Additional genotyping of a 40 cM chromosomal region surrounding the region of interest yielded a maximum parametric two-point LOD score of 1.80 (theta=0) at D2S2262 and a nonparametric LOD score of 3.11 at an adjacent novel marker 11291M1 in BAC RP11 67G7. A nonparametric multipoint LOD score of 3.20 was seen at 11291M1 under the assumption of dominant inheritance. While not providing proof of linkage considering the small number of families and large number of laboratory and statistical analyses performed, these results warrant further studies of the 2q32 chromosomal region as a candidate breast cancer susceptibility locus. Both linkage and association studies are likely to be useful, particularly in other isolated populations. PMID- 14560310 TI - Characterisation of the dysferlin skeletal muscle promoter. AB - Deficiency of the skeletal muscle membrane protein dysferlin causes the related and overlapping neuromuscular disorders limb-girdle muscular dystrophy type 2B (LGMD2B) and Miyoshi myopathy. This paper describes the preliminary characterisation of the human dysferlin promoter. The transcriptional start site of dysferlin has been mapped using 5' RACE PCR, which extended the length of the known 5' UTR to 914 bp. Promoter elements have been mapped by assessing the ability of fragments from this region to activate the expression of a luciferase reporter gene borne on a plasmid transfected into differentiated and undifferentiated C2C12 mouse myoblast cells. Finally, the core promoter region has been screened for mutations in suspected dysferlinopathy patients. PMID- 14560311 TI - Cosegregation of two unlinked mutant alleles in some cases of autosomal dominant familial exudative vitreoretinopathy. AB - Familial exudative vitreoretinopathy (FEVR) is a bilateral, clinically and genetically heterogeneous hereditary eye disorder that affects both the retina and the vitreous body. The condition has a high degree of penetrance and variable expressivity. In some cases of autosomal dominant FEVR (AD FEVR), mutations in the frizzled-4 gene (FZD-4) have been shown to be involved in FEVR pathology. In this study, we report that a second unlinked gene (Factor V) is also mutated (Leiden mutation) in the same family, which harbors the FZD-4 gene mutation. These results show for the first time that some families with FEVR could be digenic. While this is unlikely to be a widespread problem, the occurrence of digenic disorders with apparently simple Mendelian inheritance patterns renders the current method of analysis of monogenic disorders by linkage and mutation screening incomplete. PMID- 14560312 TI - Novel missense mutations in the TRPS1 transcription factor define the nuclear localization signal. AB - Deletion or mutation of the TRPS1 gene leads to the tricho-rhino-phalangeal syndromes (TRPS). The gene encodes a zinc-finger transcription factor, which contains two regions with basic amino acids LRRRRG (NLS1) and RRRTRKR (NLS2) that resemble potential nuclear localization signals (NLSs). Here, we describe the identification of novel TRPS1 mutations in patients with TRPS type I (TRPS I) and provide, by reconstructing the mutant TRPS1 proteins and subcellular localization studies, evidence that only the RRRTRKR motif functions as a NLS. Two different mutations affect the last arginine residue of this motif. The exchanges of arginine to histidine, found in two unrelated patients with TRPS I, as well as the exchange of arginine to cysteine, found in another unrelated patient, prevent the translocation of the mutant TRPS1 to the nucleus when ectopically expressed in COS 7 cells. In contrast, a mutant that lacks the conserved GATA-type zinc finger domain and most of the LRRRRG motif is able to enter the nucleus. PMID- 14560313 TI - Human X-chromosomal lineages in Europe reveal Middle Eastern and Asiatic contacts. AB - Within Europe, classical genetic markers, nuclear autosomal and Y-chromosome DNA polymorphisms display an east-west frequency gradient. This has been taken as evidence for the westward migration of Neolithic farmers from the Middle East. In contrast, most studies of mtDNA variation in Europe and the Middle East have not revealed clinal distributions. Here we report an analysis of dys44 haplotypes, consisting of 35 polymorphisms on an 8 kb segment of the dystrophin gene on Xp21, in a sample of 1203 Eurasian chromosomes. Our results do not show a significant genetic structure in Europe, though when Middle Eastern samples are included a very low but significant genetic structure, rooted in Middle Eastern heterogeneity, is observed. This structure was not correlated to either geography or language, indicating that neither of these factors are a barrier to gene flow within Europe and/or the Middle East. Spatial autocorrelation analysis did not show clinal variation from the Middle East to Europe, though an underlying and ancient east-west cline across the Eurasian continent was detected. Clines provide a strong signal of ancient major population migration(s), and we suggest that the observed cline likely resulted from an ancient, bifurcating migration out of Africa that influenced the colonizing of Europe, Asia and the Americas. Our study reveals that, in addition to settlements from the Near East, Europe has been influenced by other major population movements, such as expansion(s) from Asia, as well as by recent gene flow from within Europe and the Middle East. PMID- 14560314 TI - In NF1, CFTR, PER3, CARS and SYT7, alternatively included exons show higher conservation of surrounding intron sequences than constitutive exons. AB - It is still not fully understood to what extent intronic sequences contribute to the regulation of the different forms of alternative splicing. We are interested in the regulation of alternative cassette exon events, such as exon inclusion and exon skipping. We investigated these events by comparative genomic analysis of human and mouse in five experimentally well-characterized genes, neurofibromatosis 1 (NF1), cystic fibrosis transmembrane conductance regulator (CFTR), period 3 (PER3), cysteinyl-tRNA synthetase (CARS) and synaptotagmin 7 (SYT7). In NF1, high intron identity around the 52 constitutive and four alternatively skipped NF1 exons is restricted to the close vicinity of the exons. In contrast, we found on average high conservation of intron sequences over 300 base pairs up- and downstream of the five alternatively included NF1 exons. The investigation of alternatively included exons in CFTR, PER3, CARS and SYT7 supported this finding. In contrast, the mean intron identities around the alternatively skipped exons in CTFR and NF1 do not differ considerably from those around the constitutive exons. In these genes, the difference in intron conservation could point to a difference between the regulation of alternative exon inclusion and alternative exon skipping or constitutive exon splicing. Additional genome-wide investigations are necessary to elucidate to what extent our finding can be generalized. PMID- 14560315 TI - Comprehensive evaluation of the association between prostate cancer and genotypes/haplotypes in CYP17A1, CYP3A4, and SRD5A2. AB - Genes involved in the testosterone biosynthetic pathway - such as CYP17A1, CYP3A4, and SRD5A2 - represent strong candidates for affecting prostate cancer. Previous work has detected associations between individual variants in these three genes and prostate cancer risk and aggressiveness. To more comprehensively evaluate CYP17A1, CYP3A4, and SRD5A2, we undertook a two-phase study of the relationship between their genotypes/haplotypes and prostate cancer. Phase I of the study first searched for single-nucleotide polymorphisms (SNPs) in these genes by resequencing 24 individuals from the Coriell Polymorphism Discovery Resource, 92-110 men from prostate cancer case-control sibships, and by leveraging public databases. In all, 87 SNPs were discovered and genotyped in 276 men from case-control sibships. Those SNPs exhibiting preliminary case-control allele frequency differences, or distinguishing (ie, 'tagging') common haplotypes across the genes, were identified for further study (24 SNPs in total). In Phase II of the study, the 24 SNPs were genotyped in an additional 841 men from case control sibships. Finally, associations between genotypes/haplotypes in CYP17A1, CYP3A4, and SRD5A2 and prostate cancer were evaluated in the total case-control sample of 1117 brothers from 506 sibships. Family-based analyses detected associations between prostate cancer risk or aggressiveness and a number of CYP3A4 SNPs (P-values between 0.006 and 0.05), a CYP3A4 haplotype (P-values 0.05 and 0.009 in nonstratified and stratified analysis, respectively), and two SRD5A2 SNPs in strong linkage disequilibrium (P=0.02). Undertaking a two-phase study comprising SNP discovery, haplotype tagging, and association analyses allowed us to more fully decipher the relation between CYP17A1, CYP3A4, and SRD5A2 and prostate cancer. PMID- 14560316 TI - Phenylbutyrate increases SMN expression in vitro: relevance for treatment of spinal muscular atrophy. AB - Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disease, characterized by degeneration of the anterior horn cells of the spinal cord. SMA presents with a highly variable phenotype ranging from very severe to mild (type I-III). No cure for SMA is available at present. All forms of SMA are caused by homozygous loss of the functional survival motor neuron (SMN1) gene. However, all patients have one or more copies of the SMN2 gene, nearly identical to SMN1. Both genes encode the SMN protein but the level produced by SMN2 is insufficient to protect from disease. Increasing SMN2 gene expression could be of considerable therapeutic importance. The aim of this study was to assess whether SMN2 gene expression can be increased by 4-phenylbutyrate (PBA). Fibroblast cell cultures from 16 SMA patients affected by different clinical severities were treated with PBA, and full-length SMN2 transcripts were measured by real-time PCR. In all cell cultures, except one, PBA treatment caused an increase in full-length SMN2 transcripts, ranging from 50 to 160% in type I and from 80 to 400% in type II and III cultures. PBA was found also effective in enhancing SMN protein levels and the number of SMN-containing nuclear structures (gems). These data show that SMN expression is considerably increased by PBA, and suggest that the compound, owing also to its favorable pharmacological properties, could be a good candidate for the treatment of SMA. PMID- 14560318 TI - Challenges and opportunities in animal drug development: a regulatory perspective. PMID- 14560319 TI - Glutamate carboxypeptidase II gene expression in the human frontal and temporal lobe in schizophrenia. AB - There is decreased activity of glutamate carboxypeptidase II (GCP II) in the dorsolateral prefrontal cortex (DLPFC) and hippocampus of patients with schizophrenia. GCP II hydrolzses N-acetyl-alpha L-aspartyl-L-glutamate (NAAG), a peptide in the mammalian brain that binds to the N-methyl D-aspartate (NMDA) receptor and a group II metabotropic glutamate receptor, both of which have been implicated in the pathophysiology of schizophrenia. We examined the expression of GCP II mRNA in the DLPFC, entorhinal cortex (ERC), and hippocampus in postmortem samples from patients with schizophrenia and normal controls using in situ hybridization followed by silver grain detection. GCP II mRNA was detected in glial cells. Glial-rich regions, specifically the DLPFC and ERC white matter and the molecular and polymorphic layers in the hippocampus, express high levels of GCP II mRNA. Given the earlier finding of decreased GCP II activity in brains of subjects with schizophrenia, we expected to find lower GCP II mRNA levels in schizophrenia. Contrary to this expectation, we found a significantly higher expression of GCP II mRNA in one of the brain areas examined, the hippocampal CA3 polymorphic region. This may reflect a compensatory increase to correct for the decreased activity of GCP II activity. Our findings support the notion that the hydrolysis of NAAG is disrupted in schizophrenia and that specific anatomical regions may show discrete abnormalities in GCP II synthesis. PMID- 14560320 TI - Marijuana withdrawal in humans: effects of oral THC or divalproex. AB - Abstinence following daily marijuana use can produce a withdrawal syndrome characterized by negative mood (eg irritability, anxiety, misery), muscle pain, chills, and decreased food intake. Two placebo-controlled, within-subject studies investigated the effects of a cannabinoid agonist, delta-9-tetrahydrocannabinol (THC: Study 1), and a mood stabilizer, divalproex (Study 2), on symptoms of marijuana withdrawal. Participants (n=7/study), who were not seeking treatment for their marijuana use, reported smoking 6-10 marijuana cigarettes/day, 6-7 days/week. Study 1 was a 15-day in-patient, 5-day outpatient, 15-day in-patient design. During the in-patient phases, participants took oral THC capsules (0, 10 mg) five times/day, 1 h prior to smoking marijuana (0.00, 3.04% THC). Active and placebo marijuana were smoked on in-patient days 1-8, while only placebo marijuana was smoked on days 9-14, that is, marijuana abstinence. Placebo THC was administered each day, except during one of the abstinence phases (days 9-14), when active THC was given. Mood, psychomotor task performance, food intake, and sleep were measured. Oral THC administered during marijuana abstinence decreased ratings of 'anxious', 'miserable', 'trouble sleeping', 'chills', and marijuana craving, and reversed large decreases in food intake as compared to placebo, while producing no intoxication. Study 2 was a 58-day, outpatient/in-patient design. Participants were maintained on each divalproex dose (0, 1500 mg/day) for 29 days each. Each maintenance condition began with a 14-day outpatient phase for medication induction or clearance and continued with a 15-day in-patient phase. Divalproex decreased marijuana craving during abstinence, yet increased ratings of 'anxious', 'irritable', 'bad effect', and 'tired.' Divalproex worsened performance on psychomotor tasks, and increased food intake regardless of marijuana condition. Thus, oral THC decreased marijuana craving and withdrawal symptoms at a dose that was subjectively indistinguishable from placebo. Divalproex worsened mood and cognitive performance during marijuana abstinence. These data suggest that oral THC, but not divalproex, may be useful in the treatment of marijuana dependence. PMID- 14560321 TI - Reversal of phencyclidine-induced dopaminergic dysregulation by N-methyl-D aspartate receptor/glycine-site agonists. AB - N-methyl-D-aspartate (NMDA) receptors may play a critical role in the pathophysiology of schizophrenia. In rodents, NMDA receptor antagonists, such as phencyclidine (PCP), induce dopaminergic dysregulation that resembles the pattern observed in schizophrenia. The present study investigates the degree to which concurrent treatment with NMDA modulators, such as glycine and the recently developed glycine transport antagonist N[3-(4"-fluorophenyl)-3-(4" phenylphenoxy)propyl]sarcosine (NFPS) prevents dopaminergic dysregulation observed following chronic (3 months) or subchronic (2 weeks) PCP administration. Both chronic and subchronic treatment with PCP in the absence of glycine or NFPS led to significant potentiation of amphetamine-induced dopamine release in the prefrontal cortex and striatum, similar to that observed in schizophrenia. Treatment with either high-dose glycine or NFPS along with PCP prevented PCP effects. These findings demonstrate effective doses of glycine for use in animal models of schizophrenia, and support recent clinical studies showing the effectiveness of NMDA agonists in the treatment of persistent symptoms of schizophrenia. PMID- 14560322 TI - Childhood inattention and dysphoria and adult obesity associated with the dopamine D4 receptor gene in overeating women with seasonal affective disorder. AB - There is significant evidence that altered dopamine activity plays a role in seasonal affective disorder (SAD). The current study examined three separate genetic hypotheses for SAD related to the 7-repeat allele (7R) of the dopamine-4 receptor gene (DRD4), a variant associated with decreased affinity for dopamine. We examined the possible contribution of 7R to the overall expression of SAD, attention deficit disorder (ADD) comorbidity, and body weight regulation. As part of an ongoing genetic study of increased eating behavior and mood in female subjects, 108 women with winter SAD and carbohydrate craving/weight gain were administered the Wender-Utah Rating Scale to measure childhood ADD symptomatology, and a questionnaire to assess maximal lifetime body mass index (BMI). To test for an association between 7R and the categorical diagnosis of SAD, the transmission disequilibrium test (TDT) was used in a subsample of probands providing familial DNA. Standard parametric tests were used to compare childhood ADD symptoms and maximal lifetime BMI across the two genotypic groups defined by the presence or absence of 7R. The TDT found no initial evidence for an association between 7R and the categorical diagnosis of SAD. However, 7R carriers reported significantly greater inattention and dysphoria in childhood (p=0.01 and 0.001, respectively) and a higher maximal lifetime BMI (p=0.007) than did probands without this allele. Furthermore, excluding probands with extreme obesity (maximal BMI >40), a strong correlation was found linking childhood inattentive symptoms and maximal lifetime BMI (r=0.35, p=0.001). In overeating women with SAD, the 7R allele of DRD4 may be associated with a unique developmental trajectory characterized by attentional deficits and dysphoria in childhood and mild to moderate obesity in adulthood. This developmental course may reflect different manifestations of the same underlying vulnerability related to central dopamine dysfunction. Given the possibility of population stratification when studying genotype/phenotype relationships, future use of genomic controls and replication of our findings in other overeating and/or ADD populations are needed to confirm these initial results. PMID- 14560323 TI - In vivo evidence that 5-HT2C receptor antagonist but not agonist modulates cocaine-induced dopamine outflow in the rat nucleus accumbens and striatum. AB - During recent years, much attention has been devoted at investigating the modulatory role of central 5-HT(2C) receptors on dopamine (DA) neuron activity, and it has been proposed that these receptors modulate selectively DA exocytosis associated with increased firing of DA neurons. In the present study, using in vivo microdialysis in the nucleus accumbens (NAc) and the striatum of halothane anesthetized rats, we addressed this hypothesis by assessing the ability of 5 HT(2C) agents to modulate the increase in DA outflow induced by haloperidol and cocaine, of which the effects on DA outflow are associated or not with an increase in DA neuron firing, respectively. The intraperitoneal administration of cocaine (10-30 mg/kg) induced a dose-dependent increase in DA extracellular levels in the NAc and the striatum. The effect of 15 mg/kg cocaine was potentiated by the mixed 5-HT(2C/2B) antagonist SB 206553 (5 mg/kg i.p.) and the selective 5-HT(2C) antagonist SB 242084 (1 mg/kg i.p.) in both brain regions. The mixed 5-HT(2C/2B) agonist, Ro 60-0175 (1 mg/kg i.p.), failed to affect cocaine induced DA outflow, but reduced significantly the increase in DA outflow induced by the subcutaneous administration of 0.1 mg/kg haloperidol. The obtained results provide evidence that 5-HT(2C) receptors exert similar effects in both the NAc and the striatum, and they modulate DA exocytosis also when its increase occurs independently from an increase in DA neuron impulse activity. Furthermore, they show that 5-HT(2C) agonists, at variance with 5-HT(2C) antagonists, exert a preferential control on the impulse-stimulated release of DA. PMID- 14560324 TI - The usefulness of tilt testing with an intravenous beta-blocker in assessing the efficacy of long-term therapy in patients with vasovagal syncope. AB - BACKGROUND: Although beta blockers are frequently used for the prevention of recurrences of vaso-vagal syncope, the prediction of the long-term efficacy of this treatment is difficult. AIM: To assess whether the result of tilt testing with an intravenous beta blocker can predict the long-term efficacy of beta blockade. METHODS: The study group consisted of 62 patients (29 females, mean age 32.8+/-12.3 years and 33 males, mean age 35.9+/-18.2 years) with at least two syncopal episodes during the six months preceding positive tilt testing. After baseline tilt test, propranolol in a dose of 0.1 mg/kg body weight was administered intravenously and the tilt test was repeated. Beta blockade was defined effective if the second tilt test was negative (complete efficacy) or the time to syncope during tilt testing was increased compared with the baseline tilting (partial efficacy). All patients received chronic propranolol treatment and were followed for one year or till the recurrence of syncope. RESULTS: Intravenous propranolol prevented (n=33) or delayed (n=18) syncope during tilt testing in 51 (82%) patients whereas it was ineffective in the remaining 11 (18%) patients. During 8.6+/-6.7 (range 1-14) months of follow-up, 20 (32%) patients had syncope recurrence; 13 (25%) from the group with effective intravenous propranolol versus 7 (64%) patients in whom intravenous propranolol did not prevent syncope during tilt testing (p<0.015). Analysis of survival without a recurrence of syncope revealed a significant relationship between the results of tilt testing with intravenous propranolol and the efficacy of long-term beta blocker therapy (p<0.003). There were no significant differences between the predictive value of tilt testing with propranolol between patients with complete or partial propranolol efficacy (NS) whereas significant differences between each of these two groups and patients with ineffective intravenous propranolol were noted (p<0.04 and p<0.002, respectively). CONCLUSIONS: Propranolol administered intravenously prevents syncope during tilt testing in a significant proportion of patients. Both complete or partial efficacy of intravenous propranolol predict long-term efficacy of chronic beta blocker therapy in patients with vaso-vagal syncope. PMID- 14560325 TI - Cytokines in patients with ischaemic heart disease or myocardial infarction. AB - BACKGROUND: Cytokines are responsible for the modulation of immunological and inflammatory processes as well as proliferative responses and apoptosis. It has been recently suggested that such cytokines as interleukin 6 (IL-6), soluble interleukin 6 receptor (sIL-6R) and anti-inflammatory cytokines such as interleukin 10 (IL-10) may play a significant role in the pathogenesis of acute coronary syndromes. AIM: To assess serum concentration of IL-6, sIL-6R and IL-10 in patients with ischaemic heart disease or acute myocardial infarction (AMI). METHODS: The study group consisted of 74 patients (25 females, 49 males, aged 40 69 years) divided into three groups; group I - 18 patients with AMI (up to 12 hours from the onset of symptoms), group II - 31 patients with unstable angina and group III - 25 patients with stable angina. The control group consisted of 20 healthy subjects. RESULTS: The IL-6 and sIL-6R serum levels were significantly higher in patients from groups I and II compared with patients from group III and controls, whereas the IL-10 serum concentration was similar in all studied groups. In patients with acute coronary syndromes serum concentrations of examined cytokines were positively correlated with acute inflammatory phase parameters and classical risk factors such as body mass index, blood pressure and lipid levels. CONCLUSIONS: IL-6 and sIL-6R are markers of acute coronary syndromes and may be used for the identification of high-risk patients with unstable angina or AMI. PMID- 14560326 TI - The role of exercise ventilation in clinical evaluation and risk stratification in patients with chronic heart failure. AB - BACKGROUND: Patients with chronic heart failure (CHF) are characterised by an increased ventilatory response to exercise. The role of exercise ventilation in the risk stratification and evaluation of patients with CHF has not yet been established. AIM: To examine the relationship between exercise ventilation indices and clinical parameters of CHF and to assess the prognostic value of the ventilatory response to exercise. METHODS: The study group consisted of 87 patients with CHF (72 males, mean age 58 years) with a mean left ventricular ejection fraction of 32%. Ten patients were in NYHA class I, 38 - in NYHA class II, 34 - in NYHA class III, and 5 - in NYHA class IV. The control group consisted of 20 patients without CHF (13 males, mean age 58 years, mean LVEF - 61%). All studied subjects underwent maximal exercise test with gas-exchange measurement. The following parameters were analysed: peak exercise oxygen consumption [peak VO(2) (ml/kg/min)], VE-VCO(2) index [a coefficient of linear regression analysis depicting an association between ventilation (VE) and carbon dioxide production (VCO(2)) during exercise] and VE/VCO(2) ratio at peak exercise to VE/VCO(2) ratio while at rest (VE/VCO(2 peak/rest)). RESULTS: Ventilatory response indices were significantly higher in patients with CHF compared with controls: VE-VCO(2) - 37.9+/-11.1 vs 27.1+/-4.1; VE-VCO(2 peak/rest) - 0.89+/-0.14 vs 0.75+/-0.10 (p<0.001). In CHF patients a significant positive correlation between ventilatory response parameters and NYHA class (VE-VCO(2) - r=0.52; VE/VCO(2 peak/rest) - r=0.47) and a negative correlation with peak VO(2) (VE-VCO(2) - r=-0.52; VE/VCO(2 peak/rest) - r=-0.49) were noted (p<0.0001 for all correlations). No correlation was found between ventilatory parameters and echocardiographic variables or CHF aetiology. During the follow-up period lasting at least 12 months, 17 (22%) patients died. In the univariate Cox model, NYHA class III-IV, decreased peak VO(2) and increased VE-VCO(2) and VE/VCO(2 peak/rest) values were significantly associated with the risk of death. The multivariate analysis revealed that VE/VCO(2 peak/rest) > or =1.0 was the adverse prognostic factor, independent of peak VO(2) (p=0.02) and NYHA class (p=0.01). The Kaplan-Meier analysis showed that prognosis during the 18-month follow-up period in patients with enhanced exercise ventilation was worse than in the remaining patients (59% survival in patients with VE/VCO(2 peak/rest) > or =1.0 59% vs 91% survival in patients with VE/VCO(2 peak/rest) <1.0, p=0.001). CONCLUSIONS: In patients with stable CHF simple exercise ventilation parameters may provide important clinical and prognostic information. PMID- 14560327 TI - Factors determining long-term maintenance of sinus rhythm after cardioversion of persistent atrial fibrillation. AB - BACKGROUND: Long-term maintenance of sinus rhythm (SR) after successful cardioversion (CV) of persistent atrial fibrillation (AF) carries a low risk of stroke and may avoid the risks associated with anticoagulation. AIM: To determine the clinical and echocardiographic predictors of maintaining SR during one-year follow-up. METHODS: The initial study group consisted of 205 patients with persistent AF of whom 104 (33 females, 71 males, mean age 60.4+/-7.4 years) were randomised to SR restoration and maintenance. The results of transthoracic echocardiography, obtained before CV, were compared between patients who remained in SR and those in whom AF recurred during a one-year follow-up period, using the linear and logistic regression analysis. RESULTS: SR was present in 63.5% of patients at the end of the follow-up period. Of several analysed echocardiographic parameters, an increased left atrial area (<28 cm(2)) (p<0.02; RR 1.72, OR 1.09-2.71) and an increase in the fractional shortening of the left ventricle (range 25-40%, p<0.05, RR 1.2, OR 1.01-1.44) were significantly associated with SR maintenance during a 12-month follow-up period. CONCLUSIONS: Left atrial area and left ventricular fractional shortening are the independent predictors of the maintenance of SR after successful CV in patients with persistent AF. PMID- 14560328 TI - [In-stent thrombosis following coronary artery by-pass grafting - a case report]. AB - A case of a 60 year old male with a history of myocardial infarction treated with coronary angioplasty with stent implantation, who was admitted to the hospital for coronary artery by-pass grafting (CABG), is presented. The post-operative course was complicated by new acute MI due to the in-stent thrombosis. The patient received aspirin and ticlopidine, and underwent intra-aortic balloon pumping followed-by an effective coronary angioplasty of the infarct-related coronary artery. PMID- 14560330 TI - [Ectopic thyroid tissue in the heart - a case report]. AB - A case of a 48 year old female admitted to the hospital for further evaluation and treatment of an intracardiac tumour, is presented. The patient underwent surgical excision of the tumour localised in the right ventricular outflow tract with bioprosthetic tricuspid valve implantation and pacemaker insertion due to the procedure-related compete atrio-ventricular block. Histopathological examination revealed the presence of thyroid tissue in the excised tumour. Data published in literature dealing with this topic are reviewed. PMID- 14560331 TI - [Primary coronary angioplasty with the use of embolic protection device - a case report]. AB - A case of 65 year old male with acute myocardial infarction is described. The patient underwent successful primary coronary angioplasty with the use of a distal embolic protection device. PMID- 14560329 TI - [Massive haematoma following rescue coronary angioplasty in a patient with acute myocardial infarction - a case report]. AB - A case of a 79 year old female with acute myocardial infarction, treated with rescue coronary angioplasty, is presented. The patient received also thrombolysis, followed by heparin and IIb/IIIa platelet receptor blocker. The hospital stay was complicated by a massive groin haematoma and ecchymoses in the other parts of the lower and upper limbs, requiring blood transfusion. PMID- 14560332 TI - [Controversies regarding the effects of cyclooxygenase inhibitors on the course and complications of arterial atherosclerosis]. PMID- 14560337 TI - [Vena cava superior diverticulum - diagnostic role of echocardiography]. PMID- 14560338 TI - [Electrocardiogram of the month]. PMID- 14560339 TI - [Angiogram of the month]. PMID- 14560340 TI - [Cardioversion - induced ventricular fibrillation in a patient with pseudo-Mahaim fibers]. PMID- 14560341 TI - [Cardiotoxicity in acute poisonings - symptoms and management]. PMID- 14560345 TI - The C677T mutation in methylenetetrahydrofolate reductase gene, plasma homocysteine concentration and the risk of coronary artery disease. AB - BACKGROUND: The C677T mutation in methylenetetrahydrofolate reductase (MTHFR) gene is one of the causes of an elevated homocysteine plasma concentration and is probably one of the atherosclerotic risk factors. AIM: To assess the relationship between the presence of the MTHFR gene mutation, plasma homocysteine concentration and the risk of coronary artery disease (CAD). METHODS: The study group consisted of 120 consecutive patients (78% were male, mean age 59.2+/-9.6 years) with angiographically confirmed CAD and 106 healthy volunteers (76% were male, mean age 47.4+ or -6.0 years). The MTHFR gene mutation was detected based on the polymerase chain reaction and digestion with restrictive endonuclease HinfI. Total homocysteine plasma concentration was measured using HPLC. Folic acid and vitamin B12 plasma levels were assessed using the chemiluminescence method. Hyperhomocysteinemia was defined as homocysteine concentration > or =90 percentile of the control group which was > or =12.4 micro mol/L. RESULTS: The incidence of the mutation of allele T and the genotype TT was similar in patients and controls (51.7% vs 56.6%, and 9.2% vs 10.4%, NS, respectively). The folic acid and vitamin B12 levels were not related to the MTHFR genotype (folic acid: 8.1 ng/L in homozygotes TT vs 8.6 in heterozygotes CT and 8.3 in homozygotes CC; and vitamin B12: 273 pg/L vs 303.3 vs 314.3, respectively). Although homozygotes TT had significantly higher homocysteine concentration than heterozygotes and homozygotes CT or CC (15.4 vs 11.0 vs 11.2 micro mol/L, p<0.001), the odds ratio for CAD in genotype TT was 0.87 (95% CI 0.5-2.1, NS). The odds ratio in subjects with at least one mutated T allele was 0.82 (95%CI 0.5-1.4, NS). Homocysteine plasma concentration was significantly higher in patients with CAD than controls (12.8+/-5.1 vs 10.0+/-5.0 micro mol/L, p<0.001) and correlated significantly with folic acid (r= -0.28, p=0.0001), vitamin B12 (r= -0.19, p<0.005), age (r=0.35, p=0.0001) and creatinine (r=0.26, p=0.0001). The odds ratio for CAD in subjects with hyperhomocysteinemia was 7.1 (95%CI 3.4-14.9, p=0.001) and was 2.6 (95%CI 1.6-4.1, p=0.0001) with a homocysteine increase of 5 micro mol/L. Multivariate analysis showed that hyperhomocysteinemia was an independent risk factor of CAD (OR 2.7, 95%CI 1-7.2, p<0.05). Conclusions. Hyperhomocysteinemia rather than a mutation in the methylenetetrahydrofolate reductase gene, is an independent risk factor of coronary artery disease. PMID- 14560346 TI - Left ventricular systolic function in myocardial infarction survivors treated with primary angioplasty, thrombolysis of angioplasty preceded by thrombolysis. AB - BACKGROUND: Acute myocardial infarction (AMI) may be treated with thrombolysis, primary angioplasty or a combination of both methods. Preservation of left ventricular systolic function is an important goal of treatment. AIM: To assess whether the mode of treatment of AMI influences left ventricular systolic function measured 6 months after AMI. METHODS: In a group of 108 patients who survived AMI, an echocardiographic examination was performed 6 months afterwards. Ejection fraction, wall motion score index, asynergy area index, infarcted wall motion score index and apical segments motion score index were measured. Patients were divided into three groups: those treated with thrombolysis only, treated with angioplasty or those who underwent angioplasty preceded by thrombolysis. RESULTS: Global left ventricular systolic function was similar in all three groups. Compared to the two remaining groups, the group treated with combined therapy had significantly worse indexes of infarcted wall motion score and apical segments motion score. This group also included a significantly higher number of patients with akinetic or dyskinetic apical segments. CONCLUSIONS: Echocardiographic examination of global left ventricular systolic function in MI survivors performed 6 months after AMI, reveals similar values regardless of the method used for AMI treatment. However, segmental systolic function in the area of infarcted wall and apical segments is significantly more altered in patients treated with angioplasty preceded by thrombolysis than in other analysed patients. PMID- 14560344 TI - Rhythm control versus rate control in patients with persistent atrial fibrillation. Results of the HOT CAFE Polish Study. AB - BACKGROUND: Patients with atrial fibrillation (AF) can be managed either by maintaining sinus rhythm using antiarrhythmic drugs and/or electrical cardioversion, or by leaving patients in AF and controlling ventricular rate without attempts to restore sinus rhythm. Which of these two strategies is superior, has not yet been definitively established. AIM: HOT CAFE Polish Study (How To Treat Chronic Atrial Fibrillation) was designed to evaluate in a randomised, multicentre and prospective manner the risks and advantages of two therapeutical strategies - rate control or rhythm control, in patients with persistent AF. METHODS: The study group consisted of 205 patients (71 females and 134 males; mean age 60.8+/-11.2 years) with a mean time of AF duration of 273.7+/ 112.4 days; 101 patients were randomly assigned to rate control (Group I) whereas 104 patients were randomised to sinus rhythm (SR) restoration by DC cardioversion (CV) and subsequent antiarrhythmic drug treatment (Group II). At the end of follow-up (12 months) SR was present in 75% of patients. RESULTS: The incidence of hospital admissions was higher in group II in comparison to group I (12% vs 74%; p<0.001). Mortality was similar in both groups (1.0% versus 2.9%, NS). In both groups a significant improvement of heart failure symptoms was observed during the first 2 months (p<0.02 and p<0.001). In group II exercise tolerability measured by maximal workload during treadmill test significantly improved compared with baseline (5.2+/-5.1 vs 7.6+/-3.3 MET; p<0.0001). In patients in whom SR was restored, the left ventricular function improved and an increase in the shortening fraction was observed (29+/-7% vs 31+/-7%; p<0.01). No thromboembolic complications were observed in patients left with AF. Three patients from group II suffered ischaemic stroke; in two cases stroke was associated with CV whereas in the third patient - with late AF recurrence. CONCLUSIONS: The HOT CAFE Polish Study did not reveal significant differences in mortality between the two treatment strategies in patients with persistent AF. Although patients with SR had better improvement in some haemodynamical parameters, the hospitalisation rate was higher and the incidence of stroke was not reduced compared with the rate control group. PMID- 14560347 TI - Intraoperative assessment of mitral valve and peri-operative results of surgical treatment of mitral valve disease in patients who previously underwent closed surgical and percutaneous transluminal mitral commissurotomy. AB - BACKGROUND: Surgical open or closed mitral commisurotomy and percutaneous transluminal mitral commissurotomy (PTMC) are the well-established therapies in patients with mitral valve stenosis. However, due to various factors the long term effects may not be optimal in some patients, so they should undergo surgical mitral valve replacement. AIM: The intra-operative assessment of the morphology of mitral valve and the evaluation of the peri-operative results of surgical treatment in patients with mitral valve disease who previously underwent closed surgical commissurotomy followed by PTMC. METHODS: Twenty one patients (20 males, mean age 49 years) who underwent surgery due to mitral valve disease after closed mitral commissurotomy and PTMC, are presented. The time from closed mitral commissurotomy to PTMC was 3-42 years, and the time from PTMC to mitral valve replacement - 9 days to almost 9 years. RESULTS: One patient died on the second day after the operation because of left ventricular failure. The outcome of the remaining patients was good. Advanced changes of the mitral valve and subvalvular apparatus were present in the majority of patients. CONCLUSIONS: The results of the implantation of mitral valve prosthesis in patients who previously underwent closed surgical mitral commissurotomy and PTMC are good. In some patients with a history of closed surgical mitral commissurotomy, PTMC delays surgical replacement of the mitral valve. Advanced changes in the mitral valve leaflets and subvalvular apparatus are the causes of PTMC failure. PMID- 14560348 TI - [Iatrogenic hyperkalemia, bradyarrhythmia and the role of pacing in the elderly - three case reports]. AB - Three elderly patients with severe symptomatic bradyarrhythmia due to iatrogenic hyperkalemia are presented. In all patients potassium - lowering therapy was effective; two patients required temporary pacing. The issue of iatrogenic hyperkalemia and treatment options are discussed. PMID- 14560349 TI - [Anomalous origin of the right coronary artery from pulmonary trunk in an 11 year old girl - a case report]. AB - A case of 11 year old girl with the anomalous origin of the right coronary artery from pulmonary trunk is presented. Initially the patient was treated conservatively, however, after demonstrating myocardial ischaemia during SPECT, she was selected for surgical treatment. PMID- 14560351 TI - [Metabolic syndrome as a coronary risk factor. Prevention and treatment]. PMID- 14560350 TI - [Dilatation of superior vena cava and internal jugular vein following implantation of prosthetic mitral and aortic valves - a case report]. AB - A case of a 47 year old male with a history of closed mitral commissurotomy performed in 1991 and implantation of prosthetic mitral and aortic valves in 1999, is presented. Three years after the second procedure the patient developed dilatation of superior vena cava and internal jugular vein. PMID- 14560357 TI - [Myxoma of the pulmonary valve - a case report]. PMID- 14560358 TI - [Angiogram of the month]. PMID- 14560359 TI - [Cardiac resynchronization therapy]. PMID- 14560360 TI - [Symposium of Cardiology, Zakopane, April 11-13, 2003]. PMID- 14560363 TI - Pathological basis of gastroesophageal reflux disease. AB - Many of the present definitions of Barrett's esophagus are based on the dogma that 2 to 3 cm of cardiac mucosa normally line the distal esophagus and proximal stomach. Recent autopsy data refute this dogma. Cardiac mucosa has been shown to be frequently absent from the squamocolumnar junctional zone. When present, its extent is less than 0.5 cm in almost all children and most adults. Cardiac mucosal length increases with age. Patients who have cardiac mucosa are significantly more likely to have abnormal acid exposure in the esophagus as measured by 24-hour pH studies. The length of the cardiac mucosa correlates significantly with the amount of reflux: the greater the length of the cardiac mucosa, the more reflux there is. These new data provide insights into the pathology of gastroesophageal reflux. Normalcy is defined as an esophagus lined by squamous epithelium and a stomach lined by gastric mucosa. Reflux disease is defined by the presence of cardiac mucosa in a junctional biopsy. The severity of reflux disease is quantifiable by the length of cardiac mucosa present. Mutational reflux disease (Barrett's esophagus) is defined by the occurrence of intestinal metaplasia in cardiac mucosa and is quantitated by the amount of intestinal metaplasia present. Neoplastic reflux disease is defined as the occurrence of low grade dysplasia, high grade dysplasia, and adenocarcinoma in Barrett's esophagus. An attempt is made here to develop a rational grading system for reflux based on these highly objective histologic criteria. PMID- 14560364 TI - Effect of Helicobacter pylori infection in Barrett's esophagus and the genesis of esophageal adenocarcinoma. AB - The relation between Helicobacter pylori and gastroesophageal reflux disease is unclear. Recent reports have suggested a possible protective role for H. pylori, particularly in preventing the complications of gastroesophageal reflux disease (GERD). The purpose of this article is to present a brief overview of the recent literature regarding the role of H. pylori in the genesis of the complications of GERD, focusing on Barrett's esophagus and esophageal adenocarcinoma. The prevalence of H. pylori infection in the population of the West is around 40% and is not different in cohorts of patients with GERD. When the infection induces pangastritis or corpus-predominant gastritis, there may be concomitant reduced gastric acid secretion. Eradication of the bacteria in this subgroup of patients may enhance gastric acid secretion and provoke reflux symptoms. H. pylori organisms do not colonize the specialized intestinal metaplasia characteristic of Barrett's esophagus. H. pylori infection rates in gastric mucosa of patients with Barrett's esophagus occur at a similar or slightly lower frequency than is found in controls. Gastric infection with cagA-positive strains of H. pylori appears to be uncommon in patients with Barrett's esophagus. Furthermore, epidemiologic studies indicate that cagA-positive strains are protective against esophageal adenocarcinoma. Several investigators have proposed that the decreasing prevalence of H. pylori infection might be an important factor in the rising incidence of this tumor. PMID- 14560368 TI - Qualitative research on trauma surgery: getting beyond the numbers. AB - Qualitative research methods have a unique role to play in surgical research, revealing important contextual details about the causes and impact of surgical illnesses and treatment. The utility of qualitative methods is illustrated through results of a research study of young, black male victims of interpersonal violence. Narrative analysis of in-depth interviews with 48 survivors of penetrating violence revealed the impact of these patients' perceptions of post injury care on their pain, the central role of fear of death in their injury experience, and the transforming role providers can play in addressing patient fears of death. The study provides support for the notion that qualitative methods can be a useful approach to understanding marginalized individuals who become our patients after injury. Furthermore, potential applications of multimethod approaches to surgical research (qualitative research methods in combination with quantitative methods) for the study of trainee experiences, patient satisfaction, and racial disparities are recommended. PMID- 14560365 TI - Experimental evidence for mutagenic potential of duodenogastric juice on Barrett's esophagus. AB - The aim of this article is to review the current experimental knowledge of mutagenesis in Barrett's esophagus (BE) with special emphasis on the effect of bile salts and acid. Human evidence of direct mutagenicity is rare. Only the correlation of increased quantities and a change in the quality of bile salts with the complications of duodenogastric reflux such as BE and esophageal adenocarcinoma as an indirect marker of mutagenicity has been shown in several studies. Further evidence comes from p53 studies demonstrating an increased number of mutated p53 genes in patients with BE, esophageal adenocarcinoma, or both. Most animal and cellular experiments are carried out in a neutral pH environment, not reflecting the true nature of a reflux episode. The few studies using moderate low acid reflux conditions in combination with bile salts demonstrated a combined effect on mutagenicity. Our current knowledge of bile salt mutagenicity is predominantly based on experiments with hepatocytes and colon cancer cell lines. Future studies must be aimed at esophageal cell lines, cultured Barrett's tissue, and esophageal adenocarcinoma cell lines. PMID- 14560371 TI - Avoidance response of rainbow trout Oncorhynchus mykiss to heavy metal model mixture after long-term exposure in early development. PMID- 14560370 TI - Characterization of cholinesterases from Daphnia magna straus and their inhibition by zinc. PMID- 14560372 TI - Effects of long-term exposure of low-level diesel oil on the antioxidant defense system of fish. PMID- 14560373 TI - Growth retardation and histopathology of common carp (Cyprinus carpio) exposed to gallium. PMID- 14560374 TI - Effect of gammalin 20 (lindane) on differential white blood cell counts of the African catfish, Clarias albopunctatus. PMID- 14560375 TI - Biochemical responses of Potamogeton pectinatus L. exposed to higher concentrations of zinc. PMID- 14560376 TI - Detection of DNA strand breaks in the liver of Boleophthalmus pectinirostris treated with benzo(a)pyrene. PMID- 14560377 TI - DNA and lipid damage in the brown mussel Perna perna from a contaminated site. PMID- 14560378 TI - Coplanar polychlorinated biphenyl congeners in the liver of Galeus melastomus from different areas of the Mediterranean Sea. PMID- 14560379 TI - PH-dependent toxicity of heavy metals to a freshwater sludgeworm Tubifex tubifex Muller. PMID- 14560380 TI - Polychlorinated biphenyls and chlorinated pesticides in mussels collected from the Egyptian Mediterranean Coast. PMID- 14560381 TI - Organochlorine pesticide residues in Tana and Sabaki rivers in Kenya. PMID- 14560382 TI - Trace metal contamination in sediments of Barataria Bay, Louisiana. PMID- 14560383 TI - Pesticide residues in soil and quality of potato grown with sewage sludge. PMID- 14560384 TI - Sorption of cadmium and effects on growth, protein content, and photosynthetic pigment composition of Nasturtium officinale R. Br. and Mentha aquatica L. PMID- 14560385 TI - Growth parameters and heavy metal accumulation in poplar tree cultures (Populus euramericana) utilizing water and sludge from a sewage treatment plant. PMID- 14560386 TI - Interaction between heavy metals and nitrogen fertilizers applied to soil vegetable systems. PMID- 14560387 TI - General toxicity prediction approach for mixtures containing polar narcotic chemicals. PMID- 14560388 TI - Dissipation and offsite movement of forestry herbicides in plants of importance to Native Americans in California National Forests. PMID- 14560389 TI - Removal of nitrate contaminant in porous media aquifer through microbiological method. PMID- 14560390 TI - Biodegradation of pesticides by immobilized recombinant Escherichia coli. PMID- 14560391 TI - Efficiency of wheat bran for removal of organochlorine compounds and benzene from solution. PMID- 14560392 TI - Phytoremediation of a lead-contaminated soil using morning glory (Ipomoea lacunosa L.): effects of a synthetic chelate. PMID- 14560393 TI - Geochemistry of metals from mine tailings in Taxco, Mexico. PMID- 14560394 TI - Phthalate residues in greenhouse soil from Beijing suburbs, People's Republic of China. PMID- 14560395 TI - Insecticidal and acaricidal performance of methyl ketones in wild tomato leaves. PMID- 14560396 TI - Impact of airborne perfluorooctane sulfonate on the human body burden and the ecological system. PMID- 14560397 TI - Improved analytical procedure for determination of clopyralid in soil using gas chromatography. PMID- 14560398 TI - Temporal effect of carbofuran, a carbamate insecticide in the interruption of estrous cycle and follicular toxicity in female Swiss albino mice. PMID- 14560399 TI - Humic acid induces the expression of ox-LDL receptor in HL-60 cells through activation of PPAR-gamma. PMID- 14560400 TI - Molecular and fluorescence in situ hybridization characterization of the breakpoints in 46 large supernumerary marker 15 chromosomes reveals an unexpected level of complexity. AB - Supernumerary marker chromosomes (SMCs) of chromosome 15, designated "SMC(15)s," are the most common SMC in humans, accounting for as much as 60% of all those observed. We report the characterization of 46 large SMC(15)s, using both fluorescence in situ hybridization and polymerase chain reaction analysis within and distal to the Prader-Willi/Angelman syndrome critical region (PWACR). Our aim was to establish detailed information on origin, content, and breakpoints, to address the formation of SMC(15)s, and to facilitate genotype-phenotype correlations. For all patients in whom we were able to establish the parental origin, the SMC(15)s were maternally derived. Two patients were observed who had familial SMC(15)s, both inherited from the mother; however, in all remaining patients for whom parental samples were available, the SMC(15)s were shown to have arisen de novo. With one exception, all the SMC(15)s were shown to include the entire PWACR. Detailed investigations of the distal breakpoints categorized the SMC(15)s into two groups. Group A, representing approximately two-thirds of the SMC(15)s, had a breakpoint beyond the standard distal PWS/AS deletion breakpoint BP3, at a position close to the microsatellite marker D15S1010 and the bacterial artificial chromosome 10I10. The group B SMC(15)s were shorter, with more variable breakpoints located around BP3. The majority of the SMC(15)s were shown to have asymmetrical breakpoints, with the two inverted arms of the SMC being unequal in length. Our study revealed an unexpected level of complexity and heterogeneity among SMC(15)s that is not seen in other chromosome 15 rearrangements, such as deletions and duplications. This suggests that multiple mechanisms are involved in the formation of large SMC(15)s. PMID- 14560401 TI - Genomewide distribution of high-frequency, completely mismatching SNP haplotype pairs observed to be common across human populations. AB - Knowledge of human haplotype structure has important implications for strategies of disease-gene mapping and for understanding human evolutionary history. Many attributes of SNPs and haplotypes appear to exhibit highly nonrandom behavior, suggesting past operation of selection or other nonneutral forces. We report the exceptional abundance of a particular haplotype pattern in which two high frequency haplotypes have different alleles at every SNP site (hence the name "yin yang haplotypes"). Analysis of common haplotypes in 62 random genomic loci and 85 gene coding regions in humans shows that the proportion of the genome spanned by yin yang haplotypes is 75%-85%. Population data of 28 genomic loci in Drosophila melanogaster reveal a similar pattern. The high recurrence (>/=85%) of these haplotype patterns in four distinct human populations suggests that the yin yang haplotypes are likely to predate the African diaspora. The pattern initially appeared to suggest deep population splitting or maintenance of ancient lineages by selection; however, coalescent simulation reveals that the yin yang phenomenon can be explained by strictly neutral evolution in a well-mixed population. PMID- 14560403 TI - Implant position in knee surgery: a comparison of minimally invasive, open unicompartmental, and total knee arthroplasty. AB - This is a retrospective radiographic analysis of implant position in minimally invasive unicompartmental knee arthroplasty (UKA), open UKA, and total knee arthroplasty (TKA). Implant position and limb alignment were recorded in the AP and lateral planes. Of the 3 groups evaluated, the total knee group had the least variation and greatest accuracy of implant placement and limb alignment. UKA groups had small but significant differences in postoperative alignment and AP tibial position. Using contemporary instrumentation, UKA is less accurate than TKA in implant placement and limb alignment. Minimally invasive UKA was not as accurate as open UKA in AP tibial placement or postoperative limb alignment. PMID- 14560404 TI - All-polyethylene and metal-backed tibial components in total knee arthroplasty: a matched pair analysis of functional outcome. AB - A group of 98 patients who had primary unilateral total knee arthroplasty (TKA) with cemented, posterior-stabilized knee implants for osteoarthritis were matched for age, diagnosis, weight, body mass index, and presence of comorbid conditions. The patients were followed up with clinical, radiographic, and outcome measures including SF-36 scales and patient outcome questionnaires. Forty-nine patients had all-polyethylene tibial implants (APT), and 49 patents had metal backed tibial implants (MBT) with similar knee implant design and articular geometry. There is no difference among these patient cohorts in the clinical performance or functional outcome of TKA using APT or MBT components in primary TKA at intermediate term follow-up. In consideration of concerns about polyethylene wear, osteolysis, and cost associated with MBT components, these findings may present an opportunity for quality improvement and cost savings with increased utilization of APT in TKA operations. PMID- 14560405 TI - A two-stage approach to primary knee arthroplasty in the infected arthritic knee. AB - Fourteen patients with either septic knee arthritis or osteomyelitis of the knee with marked joint destruction were treated by resection arthroplasty with the introduction of an antibiotic cement spacer block, appropriate antibiotic therapy, and subsequent primary total knee arthroplasty. Most patients with septic arthritis had chronic infection. Eight patients had positive cultures preoperatively; the remaining patients had either a culture negative purulent aspiration or diagnostic radiographic studies. All patients underwent reconstruction with primary total knee arthroplasty on an average of 3.1 months after the initial stage procedure. Knee Society scores progressed from 46 preoperatively to 89 postoperatively, with an average range of motion from 3 degrees to 105 degrees. At latest clinical follow-up (average, 4.5 years), no patients had recurrence of infection. These data suggest that this method is successful in treating chronically infected knees with a 2-staged arthroplasty. PMID- 14560406 TI - Limited success with open debridement and retention of components in the treatment of acute Staphylococcus aureus infections after total knee arthroplasty. AB - The purpose of this study was to review our experience in treating acute gram positive infections after total knee arthroplasty with debridement, component retention, and intravenous antibiotics. Thirty-one total knee arthroplasties with acute gram-positive infections, seen at our institution over a 10-year period, were treated with open debridement and component retention. Eleven (35%) of 31 patients successfully retained their components at most recent follow-up, without ongoing infection (mean, 4 years; range, 2-10 years). Only one (8%) of the 13 patients infected with Staphylococcus aureus was successfully treated, compared with 10 (56%) of 18 patients with either Staphylococcus epidermidis or a streptococcal species. The difference between these groups is statistically significant (P=.007). The high failure rate of debridement and component retention suggests that immediate component removal should be considered in the presence of acute S. aureus infection in total knee arthroplasty. PMID- 14560407 TI - Revision total knee arthroplasty with cemented components and uncemented intramedullary stems. AB - Sixty-three failed total knee arthroplasties in 60 patients (27 females, 33 males; average age, 66 years) were treated consecutively with revision using cemented component fixation and an uncemented stem. Patients were followed for a mean of 5.75 years (range, 2-10 years); none were lost to follow-up. There were 12 (19%) re-revisions: 6 (10%) were revised for aseptic loosening, 4 (6%) for recurrent infection, and 2 (3%) for instability. Knee Society Pain Scores improved from 56 to 81, and function scores improved from 49 to 62 points. Latest radiographs in retained knees showed none with definite femoral loosening but 4 with tibial component loosening. Combining those revised for aseptic loosening and radiographic aseptic loosening, mechanical failure occurred in 10 patients (16%). PMID- 14560408 TI - Mechanical alignment of tibial stems in revision total knee arthroplasty. AB - This multicenter, retrospective study evaluates the radiographic results of achieving optimal tibial alignment in revision total knee arthroplasty (TKA) using a single modular CoCr cemented or cementless stemmed implant design. Stem size and length also were evaluated. The hundred ninety-nine revision TKAs were performed between January 1993 and January 1996 by 13 experienced revision knee surgeons. The cases were subdivided into 5 comparative groups: (1) cemented stems, (2) 140-mm length canal-filling stems, (3) 140-mm length non-canal-filling stems, (4) 95-mm length canal-filling stems, and (5) 95-mm length non-canal filling stems. The anteroposterior (AP) tibial alignment angle was measured. The canal-filling ratio (CFR) was determined by dividing the stem diameter by the endosteal diameter at the stem tip. Overall, the ability to achieve tibial alignment in the AP plane was more predictable when canal-filling (CFR >/= 0.85) cementless stems were used. This was further enhanced when long canal-filling cementless stems were selected. The least-predictable results and the highest probability of varus malalignment were achieved with cemented stems. PMID- 14560409 TI - Use of a trabecular metal patella for marked patella bone loss during revision total knee arthroplasty. AB - This study evaluates the short-term results following patellar resurfacing with a trabecular metal patella shell in the setting of marked patellar bone loss at the time of revision total knee arthroplasty (TKA). Twenty consecutive patients undergoing revision TKA with the use of a trabecular metal patella were evaluated at a mean 23-month follow-up. All patients had marked patellar bone loss at surgery precluding resurfacing with a standard cemented patellar button. Results were good or excellent in 17 of 20 patients. There were no displacements of any trabecular metal patella shells, and the fixation appeared excellent despite the poor quality of bone remaining. Complications included 3 patients with polar patella fractures postoperatively. Qualitatively, these results compare favorably with patellar resection arthroplasty in this setting. PMID- 14560410 TI - Optical analysis of surface changes on early retrievals of highly cross-linked and conventional polyethylene tibial inserts. AB - Retrieved tibial liners of highly cross-linked and conventional polyethylene were examined for articular and backside surface damage. Surfaces were graded for pitting, machine-mark loss, scratching, abrasion, delamination, and embedded debris. Whereas no difference existed in the damage score for the 2 groups, the highly crosslinked group showed significantly less elimination of machine marks. Wear, surface plastic deformation, or a combination, could account for the damage on these components. Only 1 of the highly crosslinked polyethylene inserts was available for destructive testing. That insert was melted to activate the shape memory, and thus differentiate, between wear versus plastic deformation. Nearly all changes on the articular and backside surfaces disappeared upon melting, and original machining marks reappeared, suggesting that the surface changes for that component were primarily the result of plastic deformation and not material removal. PMID- 14560411 TI - In vivo and in vitro surface changes in a highly cross-linked polyethylene. AB - Highly cross-linked, ultrahigh-molecular-weight polyethylenes (UHMWPEs) were developed to reduce UHMWPE wear in arthroplasty. These UHMWPEs have manifested an improvement in the wear resistance. Examination of the first retrievals revealed surface features not usually observed on conventional retrievals. A flattening of the machining marks is evident, together with the presence of ripples with microfissures. These ripples were investigated in vitro and on retrievals having a follow-up of up to 15 months. The examinations of all specimens showed that the ripples may be described as folds with microcracks. The depth of the microcracks extended to a maximum of 5 microm and is independent of the mode of loading (up to 27 million cycles). Because of the extreme wear resistance of these UHMWPEs, the folds accumulate on the surface of components. PMID- 14560412 TI - Clinical performance of a highly cross-linked polyethylene at two years in total hip arthroplasty: a randomized prospective trial. AB - We report the 2-year results for a prospective randomized trial comparing highly cross-linked with standard polyethylene in total hip replacements. In our study, 46 hips were available for radiographic analysis at 2- and 3-year follow-up. Femoral bearings were 28-mm cobalt chrome with the polyethylene insert randomly selected at the time of implantation to be highly cross-linked polyethylene (Crossfire; Stryker Howmedica Osteonics, Allendale, NJ) or standard polyethylene (N(2)/Vac, Stryker Howmedica Osteonics). Polyethylene wear rates were measured based on anterior-posterior (AP) and lateral pelvis radiographs at 6 weeks and at yearly intervals using a validated computer-assisted edge-detection method. Wear rates between the 2 groups were compared using the nonparametric Mann-Whitney test at the 95% level. A significant reduction in 2- and 3-dimensional linear wear rates (42% and 50%) was found in the highly cross-linked group (P =.001 and P =.005). PMID- 14560413 TI - Correlation between early and late wear rates in total hip arthroplasty with application to the performance of marathon cross-linked polyethylene liners. AB - Laboratory simulations are typically used to assess the performance of new bearing surfaces for total hip arthroplasty. However, the ability of in vitro studies to accurately predict clinical wear performance remains uncertain. Using computer-assisted radiographic measurement techniques, this study found that the average wear rate based on early (2-3 years) clinical follow-up is representative of the average long-term wear rate for a population. Based on early wear data, Marathon polyethylene liners, cross-linked with 5.0 Mrad of gamma irradiation, are wearing at a mean rate of 0.08 mm/yr. This rate is about half that of non cross-linked polyethylene but represents a more modest wear reduction than in vitro studies have predicted. If 5 to 10-year Marathon wear data corroborate our short-term results, early clinical wear data should be used to validate hip simulator studies. PMID- 14560414 TI - Degradation of mechanical properties of UHMWPE acetabular liners following long term implantation. AB - We tested the hypothesis that the mechanical and chemical behavior of gamma radiation-sterilized ultrahigh-molecular-weight polyethylene (UHMWPE) changes after implantation. Relationships between the mechanical behavior and oxidation index were explored in a cohort of 16 consecutive traceable Hexloc acetabular components (Biomet, Warsaw, IN) that were machined from extruded, stearate containing UHMWPE and gamma sterilized in air. Shelf aging time (average, 0.4 years) and implantation time (average, 11.5 years) were determined for all 16 inserts. The retrieved liners exhibited significant mechanical degradation, which was most severe in the unloaded surface regions. Analysis of the Fourier transform infrared spectroscopy data revealed a significant association between the oxidation index and mechanical degradation of the UHMWPE. The results of this study strongly support the hypothesis that the degradation of mechanical properties for the liners occurred during implantation. PMID- 14560415 TI - Twenty- to twenty-six-year radiographic review in patients 50 years of age or younger with cemented Charnley low-friction arthroplasty. AB - This is a retrospective radiographic study at 20 to 26 years of follow-up of a single surgeon's consecutive series of patients operated on at 50 years of age or younger with Charnley low-friction arthroplasty. Loosening, osteolysis, and wear were evaluated. Fifty-four patients underwent 68 low-friction arthroplasties. Fifty percent were 40 years of age or younger at surgery. The average rate of wear was 0.08 mm/yr. Osteolysis was infrequent. Twenty-two-year survival estimates and corresponding confidence levels were 76% for the cup (all causes) and 78.8% for the stem. For aseptic loosening, the figure was 80.5% for the cup and 85.2% for the stem. Successful radiographic results can be achieved in younger patients undergoing Charnley low-friction arthroplasty with more than 20 years of follow-up. PMID- 14560416 TI - Fifteen-year survivorship of a collarless, cemented, normalized femoral stem in primary hybrid total hip arthroplasty with a modified third-generation cement technique. AB - This prospective review aimed to evaluate 15-year survivorship of the collarless, third-generation cemented, normalized, Omnifit (Osteonics, Allendale, NJ) femoral stem in hybrid total hip arthroplasty (THA). Between January 1986 and June 1990, a single surgeon prospectively implanted 250 consecutive hybrid THAs (215 patients) using a modified third-generation cement technique in selected patients. A Harris-Galante (I or II) (Zimmer, Warsaw, IN) cementless shell with modular polyethylene (4150 resin) liners gamma-sterilized in air were implanted. Kaplan-Meier survivorship of the femoral or acetabular component with mechanical failure (revision for aseptic loosening) as the end-point was 100% +/- 0% at 15 years. Wear couple exchange and bone grafting was performed in 1 case (0.4%) for progressive acetabular osteolysis. This report supports femoral component centralization and good cement mantle to ensure durability of the collarless, cemented, normalized femoral stem with a surface roughness of 30-40 microinches. PMID- 14560417 TI - A prospective randomized trial of cemented femoral components with polished versus grit-blasted surface finish and identical stem geometry. AB - This randomized, prospective study compared 2 cemented hip stems that differed only in the surface finish, which was polished or grit blasted. A total of 226 hybrid total hips were evaluated at an average of 4.8 years postoperatively. No stem in either group was loose or revised for aseptic loosening. There was one case of significant distal lysis in the grit-blasted group. There was no statistical difference between polished and grit-blasted stems in incidence of lysis or bone-cement radiolucency. Harris Hip score, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), and SF-36 analysis showed no difference between groups. In conclusion, when adequate cement mantles are achieved around the femoral component, little difference in construct durability between polished and grit-blasted surface finish components can be detected at 4.8-year follow-up for this stem design. PMID- 14560418 TI - The importance of proximal cement filling of the calcar region: a biomechanical justification. AB - Clinical studies have suggested that a thicker proximal medial cement mantle improves the long-term outcomes of cemented femoral components. A 3-dimensional finite element model was used to determine the effect that replacement of proximal medial trabecular bone with cement has on cement mantle stresses. With removal of cancellous bone in the calcar region, there was a decrease in peak cement mantle stresses by approximately 20%. The greatest reduction in cement mantle stress was seen after cancellous bone was removed from the proximal medial aspect of the femur to a distance 30 mm distal to the femoral neck resection. Under fatigue loading conditions simulating gait, removal of proximal medial cancellous bone could increase the number of loading cycles to failure by a factor of 3.5. These results support the removal of proximal medial cancellous bone from the calcar region intraoperatively to increase the cement mantle thickness and reduce cement mantle stresses to improve the long-term fixation of cemented femoral components. PMID- 14560419 TI - Influence of surface finish in total hip arthroplasty. AB - The influence of femoral component surface finish was investigated by comparing 2 finishes, precoat and satin finish, for 1 cemented total hip arthroplasty (THA) system using 1 acetabular cup design. All surgeries were performed by a single surgeon in 2 consecutive series. Minimum 2-year follow-up outcomes (36 precoat, 25 satin) were compared using Harris Hip Scores, radiographs, and survivorships. The precoat population experienced significantly more radiographic and debonding failures than the satin-finish population, and significantly more pain (P <.05). Comparing failures and nonfailures within the precoat population disclosed neither significant preoperative nor significant cement-grade differences. Because of failure performance of the precoat surface finish, this surgeon no longer implants these components. PMID- 14560420 TI - Thirteen year results of total hip arthroplasty using a tapered titanium femoral component inserted without cement in patients with type C bone. AB - Although cementless arthroplasty with a tapered titanium femoral component has proven reliable in young patients with excellent bone quality, studies involving patients with poor bone quality are lacking. The present study evaluates the results of total hip arthroplasty (THA) using such a femoral component in patients with Type C femoral bone. Ninety-two THAs were performed in 81 patients aged 65 years and older using a tapered titanium cementless femoral component. Follow-up in 62 patients (72 hips) averaged 13.2 years (minimum, 10 years); 19 patients were lost to follow-up. According to Door's criteria, 20 femora were classified as Type A, 19 as Type B, and 33 as Type C. No stem was revised because of stem instability, thigh pain, or osteolysis. One stem was removed because of sepsis. Six acetabula were revised because of polyethylene wear and periacetabular osteolysis. Four patients reported mild thigh pain. Radiologic signs of osseous integration for cylindrical extensively porous coated cobalt chrome femoral components are not valid for tapered titanium designs. PMID- 14560421 TI - A comparison of cementless acetabular components of the same design: spiked versus supplemental screws. AB - The purpose of this report was to compare the results of a series of otherwise identical, cementless acetabular components with screws to a group with spikes in primary total hip arthroplasty. Between April 1993 and August 1997, 339 primary total hip arthroplasties were performed using a cementless acetabular component in 312 patients. There were 227 acetabular components with screws and 112 with spikes only. Radiographic evaluation was performed at a mean of 4.8 years' postarthroplasty (range, 2-8.6 years). No significant difference was identified between the 2 groups in regard to radiographic or clinical parameters. PMID- 14560422 TI - Cementation of an acetabular liner into a well-fixed acetabular shell during revision total hip arthroplasty. AB - Cementing a polyethylene liner into a well-fixed acetabular shell during revision hip arthroplasty may be appropriate in select patients. The purpose of this study was to review our early results with this technique. Sixteen patients with >2 year follow-up or who had any complications at <2 years from surgery were identified. Indications for this technique included: failure of the previous locking mechanism, significant polyethylene wear, malposition of the well-fixed metal shell, and previous removal of the liner with a retained acetabular shell. One acetabular shell has been revised at 13 months for multiple dislocations. Hip scores improved an average of 24.2 points. Radiographs revealed no polyethylene dissociation or acetabular loosening. Complications occurred in 7 patients. Initial results appear successful at providing secure fixation, while preventing the bone loss that might occur with removal of a well-fixed component. PMID- 14560423 TI - Use of constrained acetabular components for hip instability: an average 10-year follow-up study. AB - The short-term efficacy of using constrained acetabular components for hip instability in the total hip arthroplasty construct has been documented. However, long-term concerns including late dislocation, loosening of components, and osteolysis have not been addressed. The authors evaluated, at an average 10.3 year follow-up, 101 tripolar constrained components (Stryker Howmedica Osteonics) used in the total hip arthroplasty construct. At final follow-up, 6 hips had dislocated or had failure of the constrained component, 5 of which were revised. In addition, 4 hips were revised for aseptic loosening of the acetabular component, 4 hips were revised for aseptic femoral loosening, and 1 hip was revised for acetabular osteolysis. Considering the complexity of the cases, this component has provided durable fixation and hip stability at this intermediate term follow-up. PMID- 14560424 TI - The prevalence of corporate funding in adult lower extremity research and its correlation with reported results. AB - The author reviewed 603 consecutive articles and scientific presentations pertaining to adult lower extremity orthopedic research from 2 major American orthopedic journals and 2 major American orthopedic meetings. The prevalence of commercial funding in these studies was 50%. Clinical studies of total hip arthroplasty implants by American investigators were commercially sponsored in 75% of studies. Commercially funded hip research reported positive outcomes in 93% of studies, whereas independently funded researchers reported good results in only 37%. Funded clinical studies of total knee arthroplasty implants yielded good results in 75%; this is in contrast to the findings of independently funded investigators, who reported positive conclusions in only 20% of studies. Investigators receiving royalties reported no negative outcomes related to the respective devices. The source of research funding was strongly correlated with reported outcomes. PMID- 14560431 TI - Nurse leaders take action on range of workplace and patient safety issues. PMID- 14560430 TI - Mobile clinic 'Express' unveiled at dedication. PMID- 14560432 TI - Nursing licensure fund balances diverted to general fund. PMID- 14560433 TI - [Always under the sun. Interview by M.F. Guerel]. PMID- 14560434 TI - [Help for young diabetics: vacation counseling and prevention]. PMID- 14560435 TI - Sickle cell disease: health promotion and maintenance and the role of primary care nurse practitioners. AB - PURPOSE: To discuss the role of nurse practitioners (NPs) with regard to early identification of affected individuals, effective monitoring and screening, effective pain management and prophylaxis, and health education for patients with sickle cell disease (SCD). DATA SOURCES: Electronic database searches were performed using Medline, Cinahl, and PsycINFO. Data were obtained from medical textbooks, research, and review articles. CONCLUSIONS: SCD is a chronic inherited disease belonging to a group of conditions called hemoglobinopathies. Individuals with SCD often require close medical care from specialists. Nonetheless, NPs are in ideal positions to facilitate the health promotion and health maintenance necessary to decrease the high rate of morbidity and mortality associated with this disease. IMPLICATIONS FOR PRACTICE: NPs must understand the importance of early identification of affected individuals, effective monitoring and screening, effective monitoring and screening, effective pain treatment, and prophylaxis. The unpredictable trajectory of SCD can lead to frustration, fear, helplessness, hopelessness, and emotional distress. Ineffective pain management is a major problem for people with SCD. NPs can overcome this problem by initiating effective and prompt pain management in a nonjudgmental manner. PMID- 14560436 TI - Recognizing and treating non-infectious rhinitis. AB - PURPOSE: To increase clinicians' familiarity with nonallergic and mixed rhinitis and to differentiate these from allergic rhinitis, thus providing for an accurate diagnosis and facilitating a successful initial treatment program. DATA SOURCES: A Medline search of published journal articles was supplemented with known books and proceedings pertaining to rhinitis. CONCLUSIONS: Although there is significant overlap of symptoms among the three types of rhinitis (i.e., allergic, nonallergic, and mixed), the patient history often contains clues that can aid in establishing a correct diagnosis. The new Patient Rhinitis Screen, a questionnaire developed for use in the primary care arena, facilitates the diagnostic process. IMPLICATIONS FOR PRACTICE: As the most common condition in the outpatient practice of medicine, rhinitis is frequently treated by primary care practitioners. Recent guidelines for the diagnosis and management of rhinitis suggest that a specific diagnosis of allergic, nonallergic, or mixed rhinitis leads to more effective treatment strategies. The result is successful and efficient care utilizing, as appropriate, broad-based and symptom-specific therapies. PMID- 14560437 TI - The regulation of dietary supplements. AB - PURPOSE: To discuss the regulatory history of dietary supplements, define the term dietary supplement, clarify ingredient and nutrition information labeling, and discuss safety issues and implications for practice. DATA SOURCES: Review of primary and secondary sources, including both Internet sites and journal articles. CONCLUSIONS: In the United States, 6 out of every 10 people use dietary supplements. For decades, the Food and Drug Administration (FDA) protected the public from mislabeled and unsafe products by regulating as foods those dietary supplements that included only essential nutrients. The Nutrition Labeling and Education Act of 1990 included herbs as dietary supplements. When the Dietary Supplement and Health and Education Act (DSHEA) of 1994 was passed, the FDA lost its regulatory power. The DSHEA expanded the definition of dietary supplements beyond essential nutrients. Dietary supplements are no longer considered food additives, which makes them exempt from prescreening or any safety and efficacy studies before they are released to the public. Under the DSHEA, the FDA may take action if a product poses a direct health threat and only after adverse health effects have already occurred. IMPLICATIONS FOR PRACTICE: A good understanding of the regulatory procedures for dietary supplements will aid nurse practitioners (NPs) in patient education regarding these products. Patients should be advised to choose supplements that are made by nationally known food and drug manufacturers that belong to trade groups. NPs and patients can contact the manufacturer directly and can access government Internet sites for more product information. PMID- 14560438 TI - Evaluating the response of patients undergoing both allergy skin testing and in vitro allergy testing with the ImmunoCAP Technology System. AB - PURPOSE: To evaluate the response of patients who underwent both skin and in vitro allergy testing, both of which are accepted methods. DATA SOURCES: Retrospective review of the case notes of 100 patients evaluated by both testing methods for allergic disease. CONCLUSIONS: A total of 62 patients (62%) tested positive to at least one of the tested allergens via the in vitro method. A total of 65 patients (65%) tested positive to at least one allergen via the skin testing method. The most frequently elicited allergic response from the in vitro method was to white oak. Indoor mold and dust most frequently elicited response via skin testing. IMPLICATIONS FOR PRACTICE: Both in vitro and in vivo allergy testing have limitations. Practitioners should be aware of these when establishing a treatment plan based on the results of differing allergy testing methods. Due to differing responses to skin and in vitro testing methods, it may be prudent to perform both tests to obtain a definitive diagnosis for the allergic patient. PMID- 14560440 TI - [Outlook for control of current zoonoses in swine--an issue on consumer health protection]. AB - With the development of a uniform European strategy for the control of zoonoses the European Commission has placed the main emphasis on the protection of consumer health. This direction is clearly marked by the White Paper on Food Safety and the attached proposals for directives and regulations. The present article considers the peculiarities of the epidemiological situation of the agents Campylobacter spp. and Yersinia enterocolitica in pork production. The situation of zoonoses in Europe is used as an opportunity to present the distinctive characteristics of these agents, results of epidemiological studies available, and the risk to consumer health in light of the literature. For the approaching transfer of the responsibility for food safety to the primary producers, i.e. the farmers, the most interesting data concern the prevalence, distribution and risk factors. But as there is a strong need for clarification of further questions about the main ways of entry of these agents on farms and into the production chain before a successful preventive strategy can be developed on pig farms, these issues are considered in more detail here. PMID- 14560439 TI - Preadmission characteristics of advanced practice nursing students. AB - PURPOSE: To determine the nature of prerequisite experiences of applicants to nurse practitioner (NP) and certified registered nurse anesthetist (CRNA) programs that faculty believe are necessary for success in graduate nursing programs. DATA SOURCES: An exploratory descriptive study based on findings from four focus groups of faculty from four different universities and a written survey mailed to program or admissions directors of a random sample of 50% of the 295 credentialed NP or CRNA programs in the United States. CONCLUSIONS: Themes from the focus groups revealed 20 personal characteristics, 14 professional characteristics, and 9 clinical skills, all of which were used to create a survey instrument. A 33% response rate was achieved (n = 48). Personal characteristics were considered most important, professional characteristics and clinical skills moderately important, and kinds of clinical experience of lesser importance. Length of desired experience ranged from 0 to 5 years, most often 2 years. The most important characteristic under personal was "ethical;" under professional, "clinically competent;" under clinical, "interpersonal communication;" and under clinical experience, "to work in a setting with MD-RN interaction." IMPLICATIONS FOR PRACTICE: Information from this study may be useful for examining admissions policies of advanced practice graduate programs. Faculty were able to articulate the characteristics, proficiencies, and experiences that are likely to produce success in NP and CRNA programs. These desired characteristics need to be translated into meaningful criteria for admission. PMID- 14560441 TI - [Bacteriological findings for endodontical and apical molar dental diseases in the horse]. AB - In most cases the diagnosis of any molar dental disease in horses is made at an advanced stage, so that permanent restoration of the diseased teeth is not feasible. Complications such as bacteraemia and septicaemia due to infections as a result of dentogenous sinusitis and following dentosurgical procedures have been described in human medicine and in veterinary medicine. Twenty patients were available for examination from the Clinic for Horses of the School of Veterinary Medicine Hannover with molar dental disease in upper or lower jaw. As a result of this disease the infected tooth had to be removed surgically. The aim of this study was to determine the presence of and to identify microbes in 20 patients. Swab samples were taken from infected pulpa, from dental abscesses and from involved nasal sinuses. The samples were examined microbiologically and tested for aerobes and anaerobes at the same time. Infectious agents were found in 19 of 20 horses. In all, 27 different species of infectious agents were isolated, including both aerobic and anaerobic microbes. Fifteen patients (75%) showed a mixed flora. Further differentiation indicated a preponderance of the group of gram-negative obligatory anaerobic agents isolated from a total of 17 horses. In all these samples there was a high concentration of infectious agents of these genera, the most common of which were Prevotella spp (n = 16) and Fusobacterium spp. (n = 15). Pre-surgical antibacterial therapy is recommended to reduce the risk of intra- and/or post-surgical bacteraemia and its serious consequences. In light of these microbiological results and considering the high degree of resistance among all anaerobic microbes, all patients in this study were treated with Amoxicillin. PMID- 14560442 TI - [Diagnosis of Lawsonia intracellularis using the polymerase chain reaction (PCR) in pigs with and without diarrhea and other animal species]. AB - Lawsonia (L.) intracellularis, an obligately intracellular bacterium, causes proliferative enteropathy (PE) in swine and, occasionally, in other animals. To determine the spread of the agent among German pig herds pooled fecal samples of five animals each of clinically normal Hessian pig herds collected between november 1998 and february 1999 as well as feces (n = 1684) from individual animals representing 648 herds, sent to our laboratory by veterinarians from all parts of Germany, were tested for L. intracellularis using the polymerase chain reaction (PCR). In addition, fecal samples from diarrhoic foals (n = 46), dogs (n = 57), cats (n = 50), calves (n = 37), hedge hogs (n = 9), seals (n = 8) and one giraffe were also studied. DNA was extracted from feces using high concentrations of chaotropic salt and diatomaceous earth. For PCR, primers flanking a 279 bp fragment of L. intracellularis DNA were used (JONES, G. F., WARD, G. E., MURTAUGH, M. P., LINN, G. (1993), J. Clin. Microbiol. 31, 2611-2615). Amplificates were separated by agarose gel electrophoresis and visualized under UV-light. L. intracellularis was found in 26 (12.8%) samples from 21 (30.0%) of the Hessian pig herds without symptoms of diarrhoea. In feces of pigs with diarrhoea (n = 1684) the agent was present in 431 (25.6%) samples originating from 224 (34.6%) herds. Of the other animal species studied, L. intracellularis was detected in feces of 4 (7.0%) dogs, 2 (5.4%) calves, 3 (33.3%) hedge hogs and in the sample of the giraffe. The remaining species were all tested negative. PMID- 14560443 TI - [Possible interesting dietetic effects of lactulose as a feed additive in pig feed]. AB - In reared piglets (n = 10), fattening pigs (n = 20) and sows (n = 8) the effects of lactulose as feed additive on the quality and composition of chyme and faeces were tested to find out potential dietetic advantages of its use. The lactulose concentration in the diets varied between 27-29 g/kg dry matter in piglets and fattening pigs and 55-140 g/kg dry matter in the diet of sows. In piglets and fattening pigs the lactulose intake did not alter the dry matter content and pH in faeces; in spite of the higher concentration lactulose did not result in diarrhea in sows or adult minipigs, there was only a slight decrease in the dry matter content of faeces. In contrast to findings in human beings only traces of lactulose reached the hind gut, although higher lactulose concentrations were measured at the end of the small intestine (in piglets: 10-40 g, in fattening pigs 40-70 g lactulose per kg dry matter of chyme). The oral intake of lactulose did not increase the concentration of lactic acid or volatile fatty acids in the content of the small intestine and did not reduce the pH in the chyme, but in the contents of cecum and colon ascendens higher concentrations of volatile fatty acids were measured (in piglets: significantly higher values, in fattening pigs slightly increased concentrations). In piglets as well as in fattening pigs lactulose resulted in reduced concentrations of ammonia in the content of small and large intestine, further more the concentrations of lipopolysaccharides were increased in the colonal chyme, when pigs ingested the lactulose containing diet. Due to the fact that only small amounts of lactulose reached the hind gut there are doubts, whether lactulose has comparable dietetic effects in pigs as known from human beings (for example laxative effects, forced elimination of bacteria like salmonella). The results obtained here in pigs underline the necessity of experimental studies in the target species (including parameters of digestive physiology, chemical/microbial composition of the chyme) before substances are used and recommended for dietetic purposes in feeding practice. In spite of generally adopted similarity of the alimentary tract in pigs and human beings there are eventually some important differences within the intestine, its digestive capacity or in reactions on ingested substances leading to unexpected results. PMID- 14560444 TI - [Humoral immune response in breeding sows after vaccination with a herd-specific Chlamydophila abortus vaccine]. AB - A chlamydial vaccine efficacy trial with assessment of the clinical acceptability and serum antibody responses was performed in breeding sows. A BGM cell culture derived vaccine containing 10(8)/ml formalin-inactivated purified elementary bodies (Eb.) in sterile 0.15 M saline was prepared from Chlamydophila (Ch.) abortus strain OCHL03/99 which has been isolated in the herd from a sample of vaginal discharge. Vaccination was performed as a randomised trial with parallel treatment of a vaccinated group (25 sows) and non-vaccinated control group (20 sows). Sows received two 2.0-ml doses of vaccine intramuscularly at a three week interval. Control sows were dosed with sterile 0.15 M saline, accordingly. Serological response to vaccination was measured by ELISA with a total of 204 blood serum samples (114 from the vaccine group; 90 from the control group) using crude chlamydial LPS as the antigen. Compared to the control group, vaccinated sows showed a marked primary and secondary IgG serum antibody response following the two vaccinations. Antibody levels peaked between week 7 and 14 after priming vaccination, declined incrementally until week 27 but remained significantly higher than the corresponding sham-immune control levels and the prevaccination values of the vaccine group (p < 0.05). Western blot analysis of solubilized whole Eb. of Ch. abortus, Ch. pecorum, and Chlamydia (C.) suis with pre- and postvaccination sera confirmed that vaccination induced an antibody response preferentially against a range of 13 chlamydial antigens including the 40 kDa MOMP of Ch. abortus. Clinical side effects consisting of a transient mild local inflammatory reaction at the site of injection were observed in approx. 30% of vaccinated sows. These results provide the basis for further clinical evaluation of the Ch. abortus vaccine to protect sows from chlamydia-induced reproductive disorders. PMID- 14560445 TI - [Eradication of Prototheca zopfii infection in a dairy cattle herd]. AB - Protothecosis is a severe form of mastitis in dairy cows caused by colorless algae of the genus Prototheca. Since P. zopfii is highly resistant to all known chemotherapeutics, infected cows must be removed from the herd. Eradication measures are difficult since many chronically infected cows may become intermittent shedders. Therefore, cultural methods are insufficient for control measures. In order to eradicate Prototheca zopfii-mastitis in dairy cattle herds, two isotype specific indirect ELISA for detection of IgA and IgG1 in whey were used in a dairy herd highly affected with protothecal mastitis. All cows (n = 313) were tested four times in intervals of six months. Milk specimens were examined in parallel by cultivation and serologically using two indirect ELISA systems for specific IgA and IgG1 in whey. Cows tested Prototheca positive were consequently separated from the herd and slaughtered. At the first examination, 15.6% of the animals were found positive by culture, and 23.3% were positive in at least one of the ELISA systems. Within two years, protothecal prevalence and incidence decreased to zero indicating that the eradication strategy used was successful. In summary, serological identification of P. zopfii-infected lactating cows is an useful tool to eradicate protothecal bovine mastitis in infected herds. PMID- 14560446 TI - [Occurrence and diagnostic relevance of virulence-associated factors in Streptococcus suis]. AB - Streptococcus suis (Sc. suis) can cause very different clinical entities. In contrast to Sc. suis-associated pneumonia, the induction of meningitis, septicemia, and polyarthritis by certain Sc. suis strains requires the expression of virulence factors that contribute to the invasiveness of the pathogen. In the presented study, we examined the occurrence of known virulence-associated factors in Sc. suis isolates from samples sent to the Institute of Microbiology, School of Veterinary Medicine Hannover, in order to evaluate their significance as potential virulence factors in different disease complexes in Northern Germany. The results show that (i) MRP + EF + serotype 2 and MRP* EF-serotype 9 strains are statistically significant associated with the disease complex meningitis/septicemia/arthritis and, thus, have to be considered invasive strains, (ii) serotyping alone is not sufficient for identification of virulent strains, (iii) there is a remarkable heterogeneity among pneumonia-associated Sc. suis strains and (iv) activity of haemolysin or suilysin appears to be not appropriate as virulence marker. Finally, it has to be noted that at present only half of the Sc. suis isolates from pigs with meningitis/septicemia/poyarthritis can be characterised by the detection of virulence-associated factors. Thus, the identification and characterisation of additional, serotype independent virulence factors of Sc. suis is a very important issue in future studies. PMID- 14560447 TI - [Uncommon mycobacterial infections in domestic and zoo animals: four cases with special emphasis on pathology]. AB - Infections caused by classical tubercle bacilli are rare during the last years. Nevertheless, diseases caused by other mycobacteria have to be considered clinically and in diagnostic pathology especially in cases of immunosuppression and due to their potential zoonosis risk. An infection by mycobacteria was diagnosed in four animals (Mayotte Maki, Blue-headed Parrot, Patagonian sealion, Beagle) necropsied between 1995 and 2002 in the Institute of Veterinary-Pathology of the University of Leipzig. The Maki, the blue-headed parrot and the dog showed a disseminated character of the disease caused by Mycobacterium genavense (monkey and bird) resp. Mycobacterium avium (dog), while an open chronical tuberculosis of the lungs due to a pathogenic member of Mycobacterium tuberculosis complex was observed in the seal. All these bacteria are potential causes of zoonoses. So, if granulomatous or disseminated histiocytic alterations are detected in diagnostic pathology, mycobacterial infections should always be included in differential diagnoses and require careful aetiological investigations by histopathological and bacteriological methods. PMID- 14560448 TI - [Intestinal microecology in human and veterinary medicine--a short excursion through the human and animal gut]. AB - In honour to a great gut microbiologist the authors descend into the abdomen of humans and animals. By the way through the gut and excrements they illustrate the medical, culture-historical and literary importance of the gut and the gut flora. PMID- 14560449 TI - [Total count of bacteria in the air of three different laying hen housing systems]. AB - Bacteria in the air of animal housing is assumed to have an impact on the health of the humans and the animals in them and on the environment. The bacterial count in poultry housing systems is particularly high in comparison to those of pigs and cattle. Little is known about the bacteria in the air of new laying hen housing systems. We therefore made simple, simultaneous measurements in the air of three different systems (enriched cages, AK; conventional battery cages, BK; aviary, VOL), in the unheated scratching area of the VOL, and in the outside air over a period of one year (24-h samples were taken about once a month using polycarbonate filters) in order to determine the general bacterial count (using blood agar). The highest concentrations of bacteria were found all year long in the VOL, followed by the BK and the AK. In the VOL, there were on average 2.16 and 0.56 x 10(6) colony forming units (cfu)/m3 in the winter and summer, respectively; 0.25 and 0.38 in the BK; and 0.39 and 0.12 in the AK. These preliminary results show that air quality considerations should be included in the development and implementation of new housing systems, as should the impact on the respiratory system of the humans and animals in them and on the environment, because high concentrations of air contamination in the housing generally also lead to high emissions into the vicinity of the facility, the significance of which cannot always be estimated, as has recently been shown for antibiotics in the exhaust air from animal housing. PMID- 14560450 TI - [New family policy]. PMID- 14560451 TI - [Different methods of contraception]. PMID- 14560452 TI - [New methods of management in diabetes]. PMID- 14560453 TI - [Hormone replacement therapy and cardiology: don't dream!]. PMID- 14560454 TI - [Venous compression therapy and thromboembolism]. PMID- 14560455 TI - [Patient representative (article L. 111-6 of public health code)]. PMID- 14560456 TI - [Alendronate. Prevention and treatment of osteoporosis]. PMID- 14560457 TI - [Did you say retreat?]. PMID- 14560458 TI - Looking back. PMID- 14560459 TI - Legislative session focused on providing quality health care to Texans. PMID- 14560460 TI - Sunset law includes new requirements for dental assistants who take radiographs. PMID- 14560461 TI - DENPAC--a united voice for Texas dentists. PMID- 14560462 TI - Task force to study Medicaid challenges. PMID- 14560463 TI - Hydraulic implant abutment level impressions. PMID- 14560465 TI - To arms, to arms. PMID- 14560464 TI - Oral health of Texas residents. PMID- 14560466 TI - Oral and maxillofacial pathology case of the month. Arteriovenous malformation. PMID- 14560468 TI - Medical liability insurance; a serious burden to both patients and physicians. PMID- 14560467 TI - Localized gingival recession caused by the use of a tongue ring. PMID- 14560469 TI - The doctor is out. Medical malpractice insurance may still be a problem for many physicians, causing some to leave the profession forever. PMID- 14560470 TI - Conversations with a senator. PMID- 14560471 TI - Changing the change model: Sparks Regional Medical Center produces measurable, sustainable improvement. PMID- 14560472 TI - Using a loop configured upper arm hemodialysis graft for the "hostile" arm. PMID- 14560473 TI - Coronary artery disease during pregnancy. PMID- 14560474 TI - Campylobacteriosis: physician and laboratory practice in Arkansas. PMID- 14560475 TI - UAMS, Arkansas Department of Human Services, AMS launch nation's first 'ANGELS' program to reduce premature births. PMID- 14560476 TI - US markets for dental bone graft substitutes: executive summary. PMID- 14560477 TI - Perspectives. The ultimate end-results of implant dentistry as it is being practiced today. PMID- 14560478 TI - Apicoectomy of an endosseous implant to relieve paresthesia: a case report. AB - Various radiographic and surgical techniques have been recommended to avoid paresthesia following mandibular implant placement. However, nerve impingement is sometimes inevitable, and when lingering numbness is reported, clinicians have a limited number of corrective options. This report describes a technique for cutting-back the apex of the implant, a technique that may be useful when lingering numbness persists after osseointegration has occurred. PMID- 14560479 TI - Removal and replacement of a fractured dental implant: case report. AB - To achieve satisfactory function and esthetics following the fracture of a dental implant, the clinician is often faced with the imposing prospect of fixture removal and the restoration of adequate osseous support for subsequent fixture placement and eventual restoration. This article describes a technique that allowed removal of the fractured fixture, conservative elevation of the sinus floor and the immediate insertion of a larger diameter fixture, to permit eventual replacement. PMID- 14560480 TI - Mandibular fracture as a complication of inferior alveolar nerve transposition and placement of endosseous implants: a case report. AB - Placement of endosseous implants and inferior alveolar nerve transposition is a treatment option for patients with an edentulous posterior mandible with inadequate bone height superior to the inferior alveolar canal. Complications associated with these procedures include infection, prolonged neurosensory disturbances, and/or pathologic fracture. This report presents the surgical management of a patient with a mandible fracture after inferior alveolar nerve transposition with concurrent placement of two endosseous implants. PMID- 14560481 TI - Localized maxillary ridge augmentation with a block allograft for dental implant placement: case reports. AB - Autogenous block bone grafts have been highly successful in treating human periodontal defects, restoring esthetics, and developing adequate bone volume for dental implant placement. Limitations in available donor bone, the need for an added surgical procedure, and other potential complications have made the use of allogenic bone graft materials an important alternative. One patient described in this report presented with fractured root syndrome of the right maxillary incisor with severe resorption of the buccal plate. After atraumatic tooth extraction, a staged treatment approach involving localized ridge augmentation with an allogenic iliac bone block material and dental implant placement was used. The host bone completely incorporated the graft with only minor resorption, which enabled the implant to be placed. The allogenic bone block material used in this study was an effective alternative to harvesting and grafting autogenous bone for implant site development. The cases presented in this article clinically demonstrate the efficacy of using a block allograft in generating effective new bone fill for dental implant placement. PMID- 14560482 TI - A simple technique for fabrication of immediate interim removable prosthesis supported by transitional implants. AB - The healing period between implant insertion and exposure is often challenging for both the patient and the surgeon. The use of a removable dental prosthesis is not recommended for a period of several days to 2 weeks in an effort to avoid undue pressure being placed on the newly placed implants and on the surrounding soft tissues. However, patients are frequently uneasy without dentures, even for a short period of time, because their diet and appearance are compromised. A simple technique was developed to overcome these obstacles. This technique allows the surgeon to place an immediate implant-supported prosthesis, based on two transitional implants, in addition to the implants placed for final restoration. This technique is performed at the surgical stage simultaneously with the placement of the permanent implants. It requires minimal additional time, and makes use of the patient's original prosthesis as an interim removable restoration during the integration period. PMID- 14560483 TI - One-piece dowel-crown on single endosseous implants. AB - The unitized (one-piece) dowel-crown technique described in this article uses routine clinical and laboratory procedures, minimizes clinical appointments, and provides a simplified prosthetic restoration for a single dental implant. PMID- 14560484 TI - Crestal bone remodeling in loaded and unloaded implants and the microgap: a histologic study. AB - PURPOSE: The aim of this study was to histologically evaluate the crestal bone response to loaded and unloaded implants in beagle dogs. MATERIALS AND METHODS: Sand-blasted and acid-etched implants (Bone System, Milano, Italy) were placed in the mandible of six beagle dogs. The two premolars and the first molars had been extracted 3 months previously. Each dog received 12 implants in the mandible, and a total of 72 implants were used in this study. Three months after implantation, second-stage surgeries were performed for placement of abutments or healing screws. Three dogs were killed after 6 months, and three dogs were killed after 12 months. All 72 implants were retrieved. RESULTS: No statistically significant differences were found in the amount of bone loss between test and control implants, both at 6 and 12 months. Statistically significant differences were found, in both groups, between the bone loss observed at 6 months and that found at 12 months. CONCLUSION: Loading does not seem to be a relevant factor in the peri-implant bone resorption observed during the first year of function. Our results support previous findings that bone crest level changes could depend on the location of the microgap. PMID- 14560485 TI - Bone formation around titanium implants in the rat tibia: role of insulin. AB - BACKGROUND AND PURPOSE: Clinical and experimental studies show, with few exceptions, that type 1 diabetes mellitus is associated with a delay in bone repair around endosseous implants. The effect of insulin in bone repair/remodeling is not completely understood. The aim of this study was to investigate the course of histological and ultrastructural changes of the osseointegration process under the influence of insulin. MATERIALS AND METHODS: Titanium implants were inserted into the tibiae of male Wistar rats. Animals were divided into three groups: 1) rats with alloxan-induced diabetes; 2) diabetic rats treated with isophane insulin (2 IU/day); and 3) matching controls. Histological and histomorphometric analysis of bone-implant sections were performed 10 and 21 days after implant placement. RESULTS: Relative to control values, rats with alloxan-induced diabetes exhibited a 50% reduction in the area of formed bone (P < 0.001) and in the surface of contact between bone and implant (P < 0.01) 21 days after implant placement. There were no significant differences between groups 10 days after surgery. Values returned to normal levels in diabetic rats after insulin treatment. Presence of chondrocyte-like cells surrounded by a cartilaginouslike matrix in diabetic rats suggests a delay in the process of bone repair. Ultrastructural characteristics of bone-implant interface in diabetic rats treated with insulin resembled those observed in controls. CONCLUSION: The data presented suggest that bone repair around endosseous implants is regulated, at least in part, by insulin. The results imply that the control of the metabolic status of the diabetic patient is essential for a successful osseointegration. PMID- 14560486 TI - Radiographic evaluation of dental implants with different surface treatments: an experimental study in dogs. AB - PURPOSE: The aim of the study was to radiographically measure the bone density at the peri-implant region after osseointegration and to compare the relative bone density achieved by different surface-treated implants. MATERIALS AND METHODS: Four different types of implant surfaces were compared, using five young-adult male mongrel dogs. The first, second, third, and fourth lower premolars were extracted. Ninety days after removal, four 3.75-mm diameter and 10-mm long screw implants (Paragon) were placed with different surface treatments in the lower hemiarches. The dogs received two implants each of the following surface treatments: 1) smooth (machined); 2) titanium plasma spray: 3) hydroxyapatite coating; and 4) sandblasting with soluble particles. The implants were maintained unloaded for 90 days. After this period, the animals were killed and the hemimandibles were extracted and radiographed. The grey level of the bone adjacent to implants was measured with a specific software tool (line histogram) and the relative bone density was calculated. RESULTS: The four different surface treatments promote different numeric levels of bone density around the dental implants (sandblasting with soluble particles, 52.45 +/- 2.95; titanium plasma spray, 53.98 +/- 3.67; machined, 55.78 +/- 3.06, and hydroxyapatite coating, 58.2 +/- 2.71). Therefore, the implants can be ranked in terms of relative bone density from high to low as follows: sandblasting with soluble particles, titanium plasma spray, machined, and hydroxyapatite coating. There were no statistically significant differences in bone density among the four groups (P = 0.1130, analysis of variance). CONCLUSION: Surface treatments that add roughness to the implant show numerically higher bone density when compared with machined surfaces. The findings of radiographic density analysis suggest that the soluble blasting media-treated surface provides a greater bone density at the peri implant region. PMID- 14560488 TI - [Current state and perspectives of forensic psychiatry in Poland]. AB - The paper is an attempt to describe the current state of affairs and perspectives in Polish forensic psychiatry in relation to the EU rules and standards on dealing with psychiatric patients who break the law. Basic dilemmas and rules of forensic psychiatry are presented which allow for the formulation of a specific meta-model which in turn will enable further reforms and improvement of Polish forensic psychiatry. The author also analyses new opinion formulation rules in accordance with the criminal justica act of 1997. New legal regulations introduced solutions which improved the earlier legal regulations. This includes the role of the psychologist together with the psychiatrist, when deciding on the person's accountableness. Limiting detention use only in the cases of non accountable offenders is an unfavourable action which is against European standards and makes it difficult for many psychiatrically ill offenders to obtain therapeutic help or adequate rehabilitation. PMID- 14560487 TI - Histological analysis of the Ankylos peri-implant soft tissues in a dog model. AB - PURPOSE: The importance of the soft tissue-implant interface is enhanced by the presence of a microgap between the implant and the abutment, which represents a contamination site for bacteria. The aim of this study was to investigate the interface between the Ankylos gap-free implant system and the surrounding soft tissues in a dog model. MATERIALS AND METHODS: Six Labrador dogs were included in the study and two Ankylos implants were inserted per dog. The dogs were killed 6 months after abutment placement without functional loading and without plaque control. The implants were analysed histologically by scanning electron microscopy, light microscopy, and histomorphometry. RESULTS: Some sections exhibited histologic signs of a mild inflammation. The connective tissue between the most apical epithelial cells of the junctional epithelium and the alveolar crest was characterized by collagen fibers running from the periosteum and the alveolar crest toward the oral epithelium and, in front of the cone-shaped abutment, by a narrow zone of extracellular matrix with a few collagen fibers. CONCLUSION: Compared with results obtained in other studies using different types of implant (Astra, Branemark, ITI), the Ankylos implant showed a higher length and a larger width of connective tissue contact as well as a shorter epithelial downgrowth. The absence of a microgap in the Ankylos system could explain the histologic mild inflammation in the connective tissue. PMID- 14560489 TI - [Where do people with mental disorders are looking for help?]. AB - This article presents and discusses the concept of "pathway to psychiatric care". According to literature review, only few people, who meet criteria for mental disorders, reach psychiatrists. Therefore it is important to identify institutions where people with mental disorders look for help. On the basis of World Health Organization programme "Pathway study" authors present institutions in different countries all over the world, which are involved in helping people with mental disorders. In all European centers taken part in the "Pathway study" most of the people who reached psychiatrists had been consulted in primary healthcare first. Project "Pathway study" has been the first research programme related with this topic. It has initiated similar studies in different centers all over the world. Results of those studies show, where outside psychiatric institutions, it is possible to diagnose and treat mentally ill people. PMID- 14560491 TI - [Associative processes disorders in schizophrenia]. AB - This paper tries to approach and systematize knowledge about the character of associative process disorders in schizophrenia. In considering schizophrenia as an illness composed of various symptoms which may lead to different clinical progress, the paper is mainly focused on disorganization of thinking and, consequently, speaking and communication disorders. Authors reviewed various concept of pathogenesis and course of associative process disorders in schizophrenic patients. Special attention was paid to a connectionist model of disturbed associations. This model originates from cognitive psychology and assumes that concepts are represented as networks in the mental lexicon. Following from this model, a hypothesis was presented, claiming that disturbed associations in schizophrenia may be related to abnormalities in semantic networks. Results of research, supporting this hypothesis, were referred. Moreover, authors tried to describe the relationship between associative processes disorders in schizophrenia and abnormalities in neurophysiological (event--related potentials) and neuropathological (MRI) examinations. At least hypotheses describing the role of neurotransmission disorders was presented. PMID- 14560490 TI - [Schizophrenia in the light of the evolutionary theories]. AB - The aim of the article was to review the hypotheses of schizophrenia in the light of the evolutionary theory. The authors distinguished adaptationist and non adaptationist hypotheses concerning the evolutionary underpinnings of schizophrenia. The adaptationist hypotheses are firmly based on notions of adaptation, natural selection and proximate and ultimate causes. The standard hypotheses of this sort proposed by Steven and Price as well as by Pollimeni and Reiss were discussed. Also, the other similar conceptions originated from McGuire and Troisi and proposed by David Horrobin were presented of which the latter is especially promising and worth further investigating. The non-adaptationistic authors criticised many assumptions of the adapationist program i.e. too broad use of term "adaptation" in the area of human behaviour or the very idea of proximate and ultimate causes. Unlike the adaptionists, they focus their attention on other aspects of the evolutionary processes i.e. the role of the mutation and also they are searching in a much intensive way than their opponents for verification of the hypotheses based on the empirical evidence. The theories developed by Crow and by Yeo were listed among the non-adaptationistic ones and presented. PMID- 14560492 TI - [Neurodevelopmental theories of schizophrenia--preclinical studies]. AB - Schizophrenia is a complex disorder of unknown origin, characterised by abnormalities in the realms of perception, thinking and the experience of emotions that onset is restricted to young adulthood. Many techniques that range from neuropathology to neuroimaging identified subtle brain abnormalities particularly in frontal, temporal cortex, hippocampus, basal ganglia and cerebellum. Neurodevelopmental models of schizophrenia test hypotheses that this disease is caused by a defect in cerebral development which results in altered neural connectivity, brain neurochemistry and aberrant behaviour observed in adult life. Recent evidence indicates that neonatal hippocampal damage may affect prefrontal neuronal integrity. The developmental lesion model appears to have predictive validity because treatment with antipsychotic drugs normalises some abnormal behaviour changes. Therefore it will be a useful paradigm in the work on new therapies and in providing new insights about pathophysiology and etiology of schizophrenia. PMID- 14560493 TI - [Safety and efficacy of olanzapine versus perphenazine in patients with schizophrenia: results of multicenter, 18-week, double-blind clinical trial]. AB - AIM: The primary objective of the study was to evaluate the severity of extrapyramidal symptoms during treatment with olanzapine (10-20 mg) versus perphenazine (8-40 mg) using the Simpson Angus Scale (SAS). The secondary objective was to assess the safety profile and clinical efficacy of the investigated drugs. MATERIAL AND METHOD: A total of 95 patients with schizophrenia who met the criteria for DSM-IV were randomized to a double-blind, 18 week prospective comparative trail conducted in Poland. The tolerance of treatment was assessed with the use of scales: BAS, SAS and UKU. The efficacy of treatment was evaluated with BPRS, PANSS and CGOI scales. RESULTS: For olanzapine patients, the severity of extrapyramidal symptoms improved after 3 first weeks of treatment, and significantly decreased from the baseline to endpoint. Perphenazine patients showed an increase of extrapyramidal symptoms. The difference of the SAS scores change was statistically significant between olanzapine and perphenazine groups. Akathisia symptoms decreased significantly in the olanzapine group during the treatment period, whereas symptoms of akathisia increased in the perphenazine group. Statistically significant differences of mean change of BAS total score from baseline to endpoint were noted between treatment groups Treatment--emergent adverse events occurred more frequently in patients receiving perphenazine (46%), than in patients receiving olanzapine (17%). The proportion of patients complying with improvement criteria for CGI scale score was statistically greater in the olanzapine group (72.7%) than in the perphenazine group (47.9%). Results of this study showed that the tolerance profile in patients taking olanzapine is superior to perphenazine. CONCLUSIONS: Olanzapine was better tolerated than perphenazine. After olanzapine treatment more subjects fulfilled the criterion of improvement and schizophrenic symptoms were less severe than in patients treated with perphenazine. PMID- 14560494 TI - [Subjective aspects of pharmacotherapy in schizophrenia during implementation of chlorpromazine in Poland--a review]. AB - Introduction of chlorpromazine was the breakthrough in psychiatric practice. Pharmacotherapy with chlorpromazine started in Polish hospitals in the autumn and winter in 1954 and the first papers covering clinical experiences started to be published in 1955. Some of these papers are very informative about subjective aspects of pharmacotherapy, comparison of older and new method of treatment, changes of psychiatric wards atmosphere. They inform also about some social aspects of introduction of this new drug to the practice. In the example families were involved in gaining the drugs for ill family members and doctors sometimes had to win persons' in charge consent to start treatment. In this papers the hope that patients treated with the new drug are more prone to psychotherapy have been expressed. This kind of hope is now expressed in regard to the second generation of antipsychotics. The paper of a Polish author published 1969 that is the pioneering work in the field of research of subjective aspects of pharmacotherapy is presented. Conclusions and methodology of this study are very similar to the later studies that were published in psychiatric literature at the end of 1970's. PMID- 14560495 TI - [Comparison of subjective and objective quality of life in a group of schizophrenic patients]. AB - AIM: The aim of this study is a comparison of subjective and objective quality of life in schizophrenic patients. METHODS: The research was carried out on 120 in- and out-patients (from 19 to 65 years) who fulfilled ICD-10 and DSM-IV criteria for schizophrenia. The quality of life was assessed by means of two instruments: Quality of Life Scale (QLS, Heinrichs et al. 1984), Self-Report Quality of Life Measure for People with Schizophrenia (SQLS, Wilkinson et al. 2000). The basic statistical methods used were: correlation analysis between measurable variables with Pearson's index (P), t-Student test, analysis of variance, factor analysis. RESULTS: In the investigated group there is no correlation between subjective and objective quality of life. Sex does not influence a correlation between these variables. Subjective and objective quality of life correlate with each other only in out-patients' group (P = -0.386). The factor analysis of SQLS distinguished 7 factors: mood; everyday activities, side effects connected with movement; side effects- others; support; interpersonal contacts; others; this division differs completely from a division on subscales. The factor analysis of QLS distinguished 3 factors: intrapsychic functioning; social functioning; functioning in roles; this division is similar to a division on subscales. CONCLUSIONS: It seems that in the schizophrenic patients' group using subjective and objective ways to assess the quality of life determines a difference of the obtained results. Only the subjective measurement fulfills the assumption of quality of life definition, that is why the usefulness of QLS in assessing quality of life is rather restricted. PMID- 14560496 TI - [Role of systemic consultation in the psychiatric diagnosis]. AB - Basing on the example of one patient and her family, the advantages and disadvantages of a systemic consultation are presented. The patient was hospitalised due to schizophrenia. In the given example the psychiatrist was an observer of the consultation conducted by a therapist from the ward. A systemic consultation allowed a "multiple-picture" to be obtained: the family members and the patient presented their own understanding of the situation, their "stories" This new and multi-perspective description, showing the patient's symptoms functionality caused diagnostic uncertainty in the psychiatrist--how far is the diagnosis of schizophrenia a justifiable one. The possibility to empthasize with the existential, psychological and family problems of the patient caused a scepticism in the psychiatrist, towards the stigmatizing diagnosis. The authors stress the danger arising in the systemic consultation, which broadens and deepens the knowledge on the patient's situation. According to the authors, the systemic consultation is necessary to understand the social context of the patient and to formulate a complex treatment plan. At the same time however, the multiple layered perspective (which contains the psychological, social and existential domains of the patient) should not over-shadow the psychopathological domain. For the therapeutic team, the consideration of the given levels of systemic reality description remains a challenge. PMID- 14560497 TI - [The frequency of diagnosis of catatonic schizophrenia in hospitals in Lubliniec (1894-1932, 1934-1936, 1970-1999) and Boleslawiec (1958-1999)]. AB - AIM: The analysis of the data from the psychiatric hospitals in Lubliniec (1894 1932, 1934-1936, 1970-1999) and Boleslawiec (1958-1999) proved a decrease in frequency of diagnosis of catatonic schizophrenia, what could testify to a decrease in morbidity with this form of schizophrenia. METHOD AND RESULTS: Basing on the facts from scientific literature there was ascertained that the decrease in frequency of diagnosis of catatonic schizophrenia is connected with following causes: firstly, with the changes in nozology, secondly, with the introduction of a new form of therapy and finally, with the fact that here psychological and sociological factors exist which eliminate catatonia as the most expressive form of schizophrenia. PMID- 14560498 TI - [Revised version of the of the concept of digesting mental information]. AB - The concept of information metabolism was introduced by prof. Kepinski at the end of the 1960-ies of the 20th century. The article presents the synthesis of a series of papers aiming to concrete, update and describe in mathematical terms the concept of metabolism of information. Theoretical basis of the model in the context of the theory of information and the main structures and functions of the metabolism of information are presented. These structures are: control centre (nucleus), boundaries (cell membrane), functional structures (endoplasmic reticulum), energy centres (mitochondria), synthesis centres (ribosoms) and elimination centres (lysosoms). At the end, the publications on the mathematical description of the information metabolism model are presented and discussed. PMID- 14560499 TI - Perspectives. Part 2: Hospital finances still a guessing game for analysts. PMID- 14560500 TI - [Electroconvulsive therapy and the patient]. PMID- 14560501 TI - [Electroconvulsive therapy (ECT). Patients' evaluation of the treatment]. PMID- 14560502 TI - [Presentation of the Effort Reward Model--the new stress model]. PMID- 14560503 TI - [Selective serotonin reuptake inhibitor induced hyponatremia]. PMID- 14560504 TI - [Prevalence of child psychiatric disorders in Danish 8-9-year-old children]. PMID- 14560506 TI - [Soluble intercellular adhesion molecule-1, C-reactive protein and leukocytes pre , peri- and postpartum]. PMID- 14560505 TI - [Intrahepatic cholestasis of pregnancy treated with ursodeoxycholic acid. Results from a two-year survey at Rigshospitalet, Skejby Hospital and Odense University Hospital]. PMID- 14560507 TI - [Renal crisis at the onset of systemic sclerosis]. PMID- 14560508 TI - [Recurring urinary tract infection following an unexpected foreign body in the bladder]. PMID- 14560509 TI - [Proton pump inhibitors--are they really harmless? 1]. PMID- 14560510 TI - [Proton pump inhibitors--are they really harmless?2]. PMID- 14560511 TI - [On cancer registration]. PMID- 14560512 TI - [Assessment of articles in the Ugeskrift for Laeger by the national health research committee]. PMID- 14560515 TI - Do you work in a perfect world? PMID- 14560514 TI - History of men in nursing. PMID- 14560516 TI - Colorectal cancer. PMID- 14560519 TI - The hopeless wound. PMID- 14560518 TI - Negligent patient abandonment. PMID- 14560520 TI - Hispanics in nursing. PMID- 14560524 TI - The courage to care: intervening with colleagues who demonstrate signs of impairment. PMID- 14560522 TI - Institute for Ethics in Health Care Miami-Dade College Medical Center Campus. PMID- 14560526 TI - Time to bring Medicare into the 21st century. PMID- 14560525 TI - A case for comprehensive support for the elderly. PMID- 14560527 TI - If we keep Medicare, who pays the bills? PMID- 14560528 TI - What Medicare should look like in 2010. PMID- 14560529 TI - Testing for human papillomavirus: data driven implications for cervical neoplasia management. AB - The results of the National Cancer Institute-sponsored ASCUS-LSIL Triage Study (ALTS), the only randomized longitudinal trial in cytopathology, have provided a wealth of data to benchmark practice and stimulate discussion regarding the usefulness of human papillomavirus testing. These landmarks aided the American Society of Colposcopy and Cervical Pathology's broad-based consensus conference in integrating the Bethesda deliberations and ALTS data into clinical management. The outcomes of these open- and wide-ranging discussions have a direct impact on all who practice in this area. This article highlights the impact of these data on the contemporary practice of cytopathology and emphasizes the interaction between pathologists and clinicians. PMID- 14560530 TI - The Bethesda System 2001: an update of new terminology for gynecologic cytology. AB - From the inception of TBS in 1988, it has been well received by the pathology and clinical communities. The need for a unifying system of nomenclature was so great that most of the laboratories in the United States, as well as in numerous countries worldwide, quickly adopted TBS. By 1991, only 2 years after the initial publication of the initial TBS, 57% of United States laboratories were using the system. By 1994, almost 90% of laboratories were using TBS [28]. Although explanations for such a quick acceptance of TBS are many, one of the leading reasons is, most certainly, the sound and thoughtful basis for all of the elements of TBS. These include: (1) nomenclature that provides uniform diagnostic terminology to facilitate unambiguous communication between the laboratory and the clinician; (2) diagnostic categories that incorporate the latest scientific information on the pathogenesis and prognosis of cervical lesions; and (3) incorporation of the evaluation of specimen adequacy as an integral part of the report. From the beginning, TBS was more than just a diagnostic lexicon. When Drs. Kurman and Solomon decided that something had to be done about the diagnostic mess in GYN cytology, they did not develop another system. Instead, they convened an international consensus conference to look at the problem and come up with a comprehensive, relevant solution. The input from the entire community, which lent strength to the initial TBS, has continued to the present. This is highlighted by the enormous amount of input that was received by the 2001 TBS forum groups that enabled them to fine tune the existing terminology to meet the needs of today's clinicians. The result, continues to be an evolving, comprehensive, and clinically relevant entity, that should meet the needs of pathologists and clinicians for the foreseeable future. PMID- 14560531 TI - Atypical squamous cells: update on current concepts. AB - Besides all the confusion and associated problems that the use of ASC has created, it has initiated substantial investigational interest that has resulted in a better understanding of squamous intraepithelial lesions and the biology of cervical neoplasia. Although the category of ASC has created, and will continue to create, controversy in the diagnostic and management fields, it allows the pathologist to convey uncertainty that may be the result of poor sampling or difficulty in interpretation of a case. It is a valuable tool that the cytopathologist can use to make it known that the Papanicolaou test has its limitations and may need and benefit, in some instances, from support from ancillary studies. Similar limitations are recognized in other areas of pathology and the use of immunohistochemistry or molecular studies is widely accepted as an aid to a more specific and definitive interpretation. The time for the Papanicolaou test to be considered similarly has arrived. HPV DNA testing may not be the perfect test for cervical cancer screening because of high prevalence of HPV infection in the general population; however, it is currently the best studied ancillary test and has been proven to be cost-effective for the triage of Papanicolaou tests with equivocal squamous cells. It is important for the cytopathologist to have well-developed diagnostic skills in interpreting gynecologic preparations, and to classify cases as ASC only when deemed appropriate. Downgrading cytologic findings that are diagnostic of a squamous intraepithelial lesion to ASC with the hope of supporting it by an HPV test will only result in a devaluation of the Papanicolaou test. Such recourse may, however, be acceptable in specific situations, such as in patients who have complex histories, atypical clinical presentations, or during pregnancy. Quality assurance measures to closely monitor the ASC:SIL ratio and the rate of HPV positivity in ASC cases will be essential to ensure the appropriate use of this interpretive category. The coordination of the 2001 Bethesda and ASCCP consensus meetings resulted in the new subcategories of ASC-US and ASC-H, along with well defined management strategies for these interpretations. This new and clinically relevant terminology should lead to a reduction in difficulties at the clinical level and a more uniform management of patients, unlike the situation following Bethesda 1991 where the gynecologist was faced with a new "diagnosis" without specific management recommendations. The standardization of reporting and clinical management will also allow more reliable evaluation of patient outcomes and cost analysis. The 2002 American Cancer Society guidelines did not make specific recommendations regarding HPV DNA testing for the triage of patients who have a cytology result of ASC-US [64]. The FDA approved the expanded use of HPV testing in conjunction with the Papanicolaou test for cervical cancer screening in March of 2003. The future is likely to bring additional testing modalities that may be more specific for detecting squamous lesions that are more likely to persist or progress to carcinoma, than the currently available HPV tests. In addition, looking to the more distant future, recently published data from HPV vaccine trials suggests that immunizing women who are negative for HPV-16 may eventually reduce the incidence of cervical cancer [65]. At the present, however, the most effective method to decrease the mortality of this disease process is to make sure that all women have access to, and receive, effective cervical cytologic screening. PMID- 14560532 TI - Atypical glandular cells. AB - The 2001 Bethesda System has radically altered the classification of glandular abnormalities. The recognition of the cytologic features of atypical glandular cells on cervicovaginal smears is important because a significant number of patients will be found to have an underlying cancerous or dysplastic lesion of the exocervix, endocervix, or endometrium. The differential diagnosis of AGC on cytology is diverse and accurate classification is necessary because the most appropriate form of follow-up depends on the specific subcategorization of the atypical glandular cells. Because the level of interobserver agreement in the diagnosis of AGC is poor, effective communication between cytopathologists and clinicians is essential to accurately triage these patients. This article should help the cytology practitioner by providing a comprehensive review of the approach to the interpretation, clinical significance, histopathologic correlation, and management of patients who have atypical glandular cells on gynecologic cytology specimens. PMID- 14560533 TI - The many faces of atrophy in gynecologic cytology. AB - As more attention is paid to cervical cancer screening in the postmenopausal population, increased numbers of atrophic specimens will be evaluated in the cytology laboratory. In addition, specimens that have cell patterns that mimic the nonestrogen or partially estrogen-stimulated state occur in a variety of situations, including pregnancy, the postpartum period, and in individuals who are treated with progesterone. A firm understanding of the cellular changes that are within the range of normal in such circumstances is critical to ensure the specificity of interpretation. This article has detailed the conditions under which nonestrogen stimulated patterns occur and addressed the cytologic changes that are noted. Hints to avoid pitfalls have been offered. There is no substitute for a thorough evaluation of each case, and with continued experience and understanding of these principles, the correct interpretations, and, ultimately, correct management of patients, can be optimized. PMID- 14560534 TI - Choosing between competing technologies in the cytology laboratory. AB - Technological change often is internal to anatomic pathology services. When new technology requires collaborative development with clinical staff, it is important to systematically approach the rationale for the new technology, and be prepared to deal with its medical, financial, and, sometimes, even personal implications. Such a systematic approach involves sequentially evaluating the acceptability and the potential benefits of technologic alternatives among the laboratory leadership, with each potential vendor, the laboratory staff, and the clinical and institutional leadership, and must ultimately include effective communication with the entire clinical community. Among the benefits of such a systematic approach are resiliency of the process when challenged, and credibility of its leadership, within and outside of the laboratory. PMID- 14560535 TI - Rescreening in cervical cytology for quality control. When bad data is worse than no data or what works, what doesn't, and why. AB - The currently mandated methods to measure the sensitivity of Papanicolaou smear interpretation (including the 5-year look back and review of 10% of negative smears) are misleading. They do not allow one to measure the true sensitivity of the test and allow only a small fraction of errors to be detected and corrected. Rapid prescreening and automated screening are the only methods that seem practically feasible, and will allow the sensitivity of the method to be measured on a routine basis, and, thereby allow a reduction in overall errors. Although emerging technologies, such as HPV testing, may allow alternative methods of measuring sensitivity, the most appropriate way to use and interpret these data in this context are not yet fully developed. Unfortunately, at present there seems to be little desire to change the way things are done, and, therefore, to accurately measure sensitivity in cervical cytology. The first task that needs to be undertaken in fixing a problem is to admit that one exists. At present, most laboratory directors believe that their own laboratory is performing satisfactorily. They may well be, although the laboratories lack an analytic method to demonstrate this, and, therefore, the need for better QA methods does not seem to be acute. There is some educational value to the currently practiced and mandated performance measures, the 5-year look back and review of 10% of negative smears. Most laboratory directors seem to be happy with their QA methods and are not concerned that the data that are derived from the 10% review of negative slides does not reflect their actual sensitivity of screening in any meaningful way. Unfortunately, the forces that are currently in place in the United States ensure that accurate measures of the sensitivity of cervical cytology interpretation are unlikely to be implemented beyond the level of individual experimentation. As long as the expectation of cytologists is that the error rates are significantly less than they actually are; as long as there is significant legal and financial risk to actually measuring the true sensitivity; and as long as fictitious measures of performance are not only advocated but mandated; the confluence of incentives will ensure that the true sensitivity of the test will never be measured on a routine basis. Despite all of this, it is possible that cervical cytology screening may, in fact, already be performing at an optimal level. Being able to measure this operating performance may not effect any improvement to the overall process. The ultimate arbiter in this debate will always be the demonstration of a reduction of cervical cancer morbidity and mortality with any new measure implemented. At the present time, there is only one solution to the quality control issue; a force from outside the system must change the balance of the aforementioned incentives. The promise of data from European experiences with rapid rescreening may show that this method is effective and accurate. Such data might make the current methods that are in use in the United States more open to change. So yes, the answer is that "bad" data may be worse than no data at all. The bad data that we have been collecting for more than a decade is as effective a trap as anyone could have devised to ensure that actually measuring the performance of cervical smear interpretation does not happen. The only question that remains is, "How we will be able to escape?" PMID- 14560536 TI - Molecular markers in cervical cytology. AB - It is expected that in the near future, the high sensitivity of array-based technologies and identification of panels of molecular fingerprints that are specific for each disease process will allow the pathologist to analyze cytologic samples and tissue biopsies by these technologies in conjunction with morphologic evaluation. This approach could lead to a new era in diagnosis and patient management, where each patient may receive individualized treatment according to the molecular characteristics of the disease that are obtained from a minute amount of tissue. Therefore, it is important for pathologists and other clinical specialists to have an understanding of these molecular technologies. It is hoped that this article will allow practitioners to incorporate these concepts into their training, and, eventually, into their daily practice. PMID- 14560537 TI - Effusion cytology in ovarian cancer: new molecular methods as aids to diagnosis and prognosis. AB - Carcinoma of the ovary is the leading cause of death from gynecologic cancer in western countries. Ovarian carcinoma is commonly associated with the accumulation of fluid that contains malignant cells in the peritoneal and pleural cavities. This article details the current knowledge regarding the diagnostic and biologic characteristics of ovarian carcinoma cells in effusions, and the genotypic and phenotypic differences between solid primary tumors and metastatic lesions. Finally, we present a new approach, by which the analysis of fresh frozen viable cells allows us to study in vivo the links between expression and activity of extracellular mediators, membrane receptors, intracellular signaling, and transcription factors, and their potential therapeutic and prognostic significance. PMID- 14560538 TI - Cervical cytology automation: an update for 2003. The end of the quest nears? AB - The field of cytology automation, through long investigation, trial and error, and finally, commercial success and failure, has arrived at the first levels of the "grail" of improvements in accuracy and productivity in cervical cytology screening. It remains to be seen how much further the road will lead toward so called "diagnostic" instrumentation that would actually provide us with a fully automated system of "specimen in-diagnosis out" with little, or no, human input. Will commercial ventures or academic institutions continue to support investigations to further the applications that have been developed to date? This remains to be seen and is directly dependent on parallel processes that are detailed elsewhere in this issue. Will HPV vaccines eliminate the need for screening? Possibly, but probably not for many years [70]. Will more sensitive and specific genetic or protein markers (or combinations thereof) be found to be more accurate and cost-effective? Certainly the possibility of mass screening by high-risk HPV DNA testing, as a viable alternative, is being discussed at present. Despite all of these uncertainties, the present (or nearly available) technology has the potential to improve the practice of cervical cytology. Improvements in accuracy that are necessary to provide the highest possible level of patient care and to protect practitioners from unreasonable levels of medico legal risk are a reality. Improvements in productivity that are necessary to help in the impending labor shortage in the field of cytotechnology are also a reality. Automation is clearly the short-term solution to the most difficult of the challenges that we face. PMID- 14560539 TI - Bonding of resin to dentin. Interactions between materials, substrate and operators. AB - OBJECTIVES: The general aim of this thesis was to identify and study factors that affect bonding between resin and dentin, including operator variability. MATERIALS AND METHODS: Bonding sites were generated in vivo and in vitro and compared using SEM. The effects of experimental water- or acetone-based primers on shear bond strength to dentin were studied in vitro. Fourier transform infrared spectroscopy was used to establish any detrimental effects from water on polymerization by determining the degree of conversion of thin films of resin. The interaction between phosphoric acid and dentin was analyzed by measuring calcium leaching by use of atomic absorption spectroscopy. Bond strength of composite resin was measured to dentin with various degrees of demineralization. Gap formation adjacent to composite restorations in standardized dentin cavities was studied in vitro by confocal microscopy. RESULTS: A similar morphological appearance was found for bonding sites generated in vitro vs. in vivo. An acetone based primer was more dependent on a moist bonding technique than was a water based system. However, water might influence bonding by interfering with the polymerization of the resin. Calcium leaching from dentin can be predicted by use of a solubility phase diagram. No correlation could be established between calcium leaching and bond strength. Gap formation was more dependent on the operator than the choice of material. A simplified all-in-one adhesive showed less operator variability compared to more complex bonding systems. CONCLUSIONS: In vitro bond testing may well indicate the outcome of in vivo trials. The solvents used in bonding agents will influence the performance and, presumably, technique sensitivity. Water rinse time after etching might be a more important consideration than etch time. In spite of the relative importance of the results presented above, the outcome of the multi-operator trial could not be predicted. PMID- 14560540 TI - Types and epidemiology of tendinopathy. AB - During the last few decades, the role of sports and physical activity has become more and more important in all modern communities. The risk of tendon injury has thus increased, and prevention has become important. Epidemiologic studies are important when planning prevention programs for tendon injuries. Because of individual sport cultures and different sport habits in different countries, national epidemiologic studies are of importance in each individual country. PMID- 14560541 TI - Cell death and tendinopathy. AB - Apoptosis and necrosis are presently recognized as the two major types of physiological and pathological cell death. Apoptosis is a tightly regulated cell deletion process that differs morphologically and biochemically from necrotic cell death. Tendinopathy is defined as a tendon injury that originates from intrinsic and extrinsic etiological factors. Excessive apoptosis has recently been described in degenerative tendon. The increased number of apoptotic tendon cells in degenerative tendon tissue could affect the rate of collagen synthesis and repair. Impaired or dysfunctional protein synthesis may lead to weaker tendon tissue and eventually increase the risk for tendon rupture. Clearly, there are many details to insert into this pathway, but there is hope that if the fine details of the pathway can be fleshed out, then strategies may be able to be developed to break the cycle at one or more points and prevent or treat tendinopathy more effectively. PMID- 14560542 TI - Compression etiology in tendinopathy. AB - Recent studies have emphasized that the etiology of tendinopathy is not as simple as was once thought. The etiology is likely to be multifactorial. Etiologic factors may include some of the traditional factors such as overuse, inflexibility, and equipment problems; however, other factors need to be considered as well, such as age-related tendon degeneration and biomechanical considerations as outlined in this article. More research is needed to determine the significance of stress-shielding and compression in tendinopathy. If they are confirmed to play a role, this finding may significantly alter our approach in both prevention and in treatment through exercise therapy. The current biomechanical studies indicate that certain joint positions are more likely to place tensile stress on the area of the tendon commonly affected by tendinopathy. These joint positions seem to be different than the traditional positions for stretching exercises used for prevention and rehabilitation of tendinopathic conditions. Incorporation of different joint positions during stretching exercises may exert more uniform, controlled tensile stress on these affected areas of the tendon and avoid stresshielding. These exercises may be able to better maintain the mechanical strength of that region of the tendon and thereby avoid injury. Alternatively, they could more uniformly stress a healing area of the tendon in a controlled manner, and thereby stimulate healing once an injury has occurred. Additional work will have to prove if a change in rehabilitation exercises is more efficacious that current techniques. PMID- 14560543 TI - The painful nonruptured tendon: clinical aspects. AB - Tendon conditions cause a great deal of morbidity in both elite and recreational athletes, and outcome of treatment is often unsatisfactory. Evidence that the common clinical conditions (e.g., Achilles, patellar, elbow and rotator cuff tendinopathies) are due to tendinosis has been present for many years, yet the misnomer "tendinitis" is still widely used for these conditions in clinical practice. Clinical practice remains very different from evidence-based recommendations [8], but this is a common challenge in medical practice. Thus, in addition to further research in an area of medicine rife for such endeavor, there must be attention to knowledge translation--ensuring that the patient benefits from what is already known. PMID- 14560544 TI - Chronic midportion Achilles tendinopathy: an update on research and treatment. AB - Chronic painful midportion Achilles tendinosis has been known as a difficult condition to treat, and surgical treatment was often needed. In recent scientific studies, however, treatment with painful eccentric calf muscle training has shown very good short-term clinical results and significantly reduced the need for surgical treatment. Also, very recently, a new method consisting of sclerosing of neovessels in the area with painful tendinosis showed promising short-term results. Ongoing and future research will evaluate the potential of these treatment models. PMID- 14560545 TI - Patellar tendinopathy. AB - Patellar tendinopathy is a common and serious condition in athletes. Although there have been many advances in the understanding of the histopathology, imaging, and surgical outcomes in this condition in the past decade, successful management of athletes with patellar tendinopathy remains a major challenge for both the practitioner and patient. There is a definite need for further prospective studies into etiological factors and randomized controlled trials into treatment choices. PMID- 14560546 TI - Rupture of the Achilles and patellar tendons. AB - Certain similarities can clearly be appreciated between Achilles and patellar tendon ruptures. Both are strong tendons that transmit force bridging at least one joint of the lower limb. When healthy, both require massive forces to be disrupted, and both can be weakened through certain systemic disease processes, steroids, and fluoroquinones. Both allow for a variety of innovative management possibilities that ultimately lend themselves to individual surgical preference. We feel that, although surgical management plays an important role in restoring continuity in knee extension and in plantar flexion, functional outcome inevitably relies on patient motivation and a well-established physiotherapy regime. Sports physicians should be able to identify both conditions early in their presentation, but still hold a high index of suspicion for these problems in athletes who have an acute exacerbation of ongoing tendinopathy. PMID- 14560547 TI - Rehabilitation of lower limb tendinopathies. AB - Tendon rehabilitation requires an extensive assessment, and identification of deficits that impact on sporting performance. Rehabilitation must address all these deficits and may take several months. Return to sport may require short- or long-term load modification. Surgery also requires a long recovery time, and may not guarantee a return to sport. PMID- 14560548 TI - Etiologic and pathogenetic factors for rotator cuff tendinopathy. AB - Etiologic and pathogenetic factors for rotator cuff tendinopathy, although often compartmentalized to intrinsic or extrinsic causes, have multifactorial roots. The development of animal models for the study of rotator cuff disease has increased the fund of knowledge regarding this disease and has paved the way for future studies. Further multidisciplinary studies at molecular, biomechanical, and clinical levels should be undertaken to enhance the understanding of this common disorder. Ultimately, the goals of improved care, increased comprehension, and prevention of rotator cuff tendinopathy are attainable. PMID- 14560549 TI - Elbow tendinopathy: tennis elbow. AB - The pathoanatomy of overuse tendinopathy is noninflammatory angiofibroblastic tendinosis. The areas of elbow abnormality are specific, including the ECRB-EDC complex laterally, the pronator teres, flexor carpi radialis medially, and triceps posteriorly. The goals of nonoperative treatment are to revitalize the unhealthy pain-producing tendinosis tissue. The key to nonoperative treatment is rehabilitative resistance exercise with progression of the exercise program. If rehabilitation fails, the surgical interventions as described are highly successful. PMID- 14560550 TI - Rehabilitation of rotator cuff tendinopathy. AB - Rehabilitation of the dysfunction that is associated with rotator cuff tendinopathy should be based on the evidence known about the pathoetiology of the tendinopathy, what is known about the extent of the local anatomic injury, the local and distant physiological and biomechanical alterations, and on the knowledge developed regarding progressive loading of the injured or altered structures. Prehabilitation, or prospective exercises to minimize future rotator cuff loading stresses, should be included at the end of rehabilitation as part of the return to function. PMID- 14560551 TI - [From physics to biology: the intrinsic dynamics of the cosmos]. AB - The History of Universe is described in an extremely summarized manner through the use of graphics, from Big bang until today. This is done according to the most recent standard models of Cosmology and Physics of elementary particles; in other words, according to those fields in Physics of a presupposed universal reference. The History of Life is immersed in this universal physical context, in a frame where our knowledge from Geology and Biology can be only terrestrial. The underlying ideas we try to arise are: 1) the transition from a relatively elementary structure to a posterior and a relatively more complex one requires some very special "environmental" conditions; and 2) the new structure can not be described only through its materic constituents, because in cosmic dynamicity new structures and new relationships (of intrinsic respectivity) arise, together with new laws (of extrinsic respectivity). Consequently and as an objective, physical knowledge (for example, elementary particles or atoms) alone in no way can explain biological reality (for example, cell or man). PMID- 14560552 TI - [Transplantation of cerebral neurons into spinal cord for the treatment of paraplegia]. AB - This report shows the experimental results obtained after microsurgical cotransplantation of fetal cerebral tissue and peripheral nerve tissue into previously injured spinal cord. Between 8 and 12 months after surgery, functional recovery is observed in the grafted animals, associated with an increase in muscle mass in the lower extremities. At this time, donor cerebral tissue is integrated into previously injured spinal cord and results in formation of bundles of nerve fibers that emerge from the area of the transplant and surround the spinal cord beneath the lesion. These findings support the possibility of functional recovery after chronic traumatic paraplegia. PMID- 14560553 TI - [Eugenio Espejo, the Quito physician: pioneer of bacteriology in the Americas]. AB - Eugenio Espejo (1747-1795). Mestizo. Was born in the Royal Audience of Quito. Today, the Republic of Ecuador. The 18th century was isf illustration and enlightment for the Quitenos. Espejo's extraordinary erudition is explained by the full access he had to yhe libraries created in Quito by the Jesuits. The best in the continent at that time. His work, Reflections on Smallpox, makes or Espejo outstanding in his field. Pioneer in bacteriology and of the biopathological observations in America. The first newspaper in Quito, Primicies of the culture of Quito, was his own work. Furthermore, Espejo is considered the precursor of the independence of Hispanoamerica. PMID- 14560554 TI - [Development of resistance and antibiotic consumption]. AB - The development of antibiotics resistance is reaching epidemic proportions. Due to increased transportation of persons and goods across continents, some bacterial clones have disseminated over several countries. To increase benefits and reduce risk, and therefore improve quality of prescription, the rational use of antibiotics should be promoted. To do so, patterns of use and prescription rates expressed in DDD per 1,000 hospital beds should be examined. Hospitalised patients received 25-40% of all systemic doses, in many cases inappropriately. In community studies reference estimates are made per 1,000 inhabitants-day (DHD). Over the last years, consumption has levelled-of around 20 DHD in 2000, showing remarkable differences between regions in Spain. Antibiotic consumption is approximately 270 Tons (in 1997), with an increase of only 10% in the last decade. The URANO study estimated that 19% of patients receive chemotherapy, although 42% do not comply with treatment and 29% are self-medicated. Among the chemotherapeutic agents mostly used are penicillins (11.4 DHD), macrolides (3.6 DHD), Quinolones (2.2 DHD) and Cephalosporins (1.9 DHD). We recommend surveillance of antibiotics resistance and improvement in the training of health professionals and in health information for patients about risks of self medication; we also propose a better control of antibiotics administered to farm animals. PMID- 14560555 TI - [Vaccination of adolescents and adults]. AB - Today nobody doubts about the need of maintaining adults and teenagers dully vaccinated, among other reasons because they had not been vaccinated when they were young, due to the need of a booster dose, or due to the availability of new vaccines. Several international organisms recommend the vaccination to teenagers and adults bearing in mind several factors, such as the type of vaccine, age, sex, occupation and medical situation of the person to be vaccinated. The recommended vaccines will produce a positive effect, reducing the morbi-mortality rates among the vaccinated. PMID- 14560557 TI - [3 summit conferences on Earth insufficient to save the Planet, Controversies and achievements]. AB - This review summarizes the present concerns on the deteriorated biological systems. This concern has generated many publications since it is a very serious subject. The human being has produced a deep impact on the environment that could even could induce climatic changes. Because these circumstances have increased the socioeconomical differences between developed and third world countries, the United Nations Organization had organized three summit conferences (Oslo, 1972; Rio do Janeiro, 1992; Johannesburgh, 2002) in order to propose solutions. This review discusses the conclusions held at these conferences. PMID- 14560556 TI - [Clinical prediction in medical oncology]. AB - Predictive factors (PF) are variables that give information about survival, treatment response or toxicity and future complications on cancer patients. The foremost utility of PF takes root in the possibility to furnish an individualized treatment schedule with higher succeeding options. They include clinical characteristics of the patient, tumour features, treatment administrated, classical pathological and new molecular data obtained from patients clinical samples. Clinical parameters comprise age, sex, underlying diseases and performance status among others, and in concurrence with tumour pathology and clinical stage (TNM) usually define the best treatment options. Also, chemotherapy response can modify natural history of several tumours, and thus is a PF. Modifications in evolving PF typically induce a variation in patient outcome. Hence, surgical tumour size reduction or neoadjuvant down-staging improve survival in several cancers. In the other side, treatment adjustment to steady PF should offers better outcome than "standard therapies". Recent advances on cancer research have generated a great deal of biological data that help us to search new treatment and diagnostic modalities. Biotechnology offers a great amount of possibilities in the next future and probably a true individualized therapy. Conversely, there are a small amount of molecular evidences that imply a creal variation in current clinical practice. Hence, more scientific and financial efforts are necessary to exploit to the full knowledge spurting up from basic science. In summary, the prediction in oncology is a hard task derived from clinical observation, tumour behaviour, treatment schedules and biological evidences that must offer realistic predictions on a concrete cancer patient. Oncologists have a duty to know all these variables to accomplish this thorny assignment. This review will focus on classical and recent biological PF in cancer. PMID- 14560558 TI - [Health and social challenges originated by the aging of the population]. AB - The increment in ageing ratios, mainly in developed countries, and the increment in life expectancy, 83 years-of-age for Spanish women, have created a new caring scenery in health care confronting the "Geriatrisation of Medicine" and social care dealing with systems covering the needs of an aged population pertaining to a society whose family nucleus meets increasing difficulties for attending their elderly members. An efficacious geriatric care system should be based on solid hospital care, community oriented care and integration of health and social care, always aimed towards functional recovery of the individual and the integration of elderly people in their communities. PMID- 14560559 TI - HPV testing. Visible expectations and hidden realities. PMID- 14560560 TI - Risk factors for local recurrence in patients with invasive breast cancer and negative surgical margins of excision. Where are we and where are we going? PMID- 14560561 TI - Efficiency of the hybrid capture 2 HPV DNA test in cervical cancer screening. A study by the French Society of Clinical Cytology. AB - The aim of this study was to determine the efficiency of the Hybrid Capture 2 (HC2; Digene, Gaithersburg, MD) human papillomavirus (HPV) assay for the detection of cervical neoplasia. Of the 1,785 patients recruited, 462 (25.88%) were referred for colposcopy owing to previously detected cytologic abnormalities, and 1,323 (74.12%) were voluntary candidates for screening. For all patients, a Papanicolaou smear and a monolayer smear (ThinPrep, Cytyc, Boxborough, MA) were done. HPV DNA was detected on the residual liquid-based material. False-positive results were observed in 111 cases and comprised 34 cross-reactions (1.90%) and 77 false-positive cases (4.31%) owing to a contiguous strong chemiluminescence signal. Interestingly, all these samples had a relative light units value of 1 to 3 and were contiguous to a sample with a very high HPV DNA load. The final results showed that high-risk and low-risk HPV DNA were detected in 480 samples (26.89%) and 135 samples (7.56%), respectively. Although HC2 can be considered a reliable and sensitive test for HPV DNA detection, we do not advocate its use for large-scale screening for cervical neoplasia. PMID- 14560562 TI - Factors associated with ipsilateral breast failure and distant metastases in patients with invasive breast carcinoma treated with breast-conserving therapy. A clinicopathologic study of 607 neoplasms from 583 patients. AB - All slides from 607 consecutive invasive breast carcinomas from 583 patients were reviewed. Margin distance, composite width of invasive carcinoma, and number of ducts with intraductal carcinoma within the one-half low-power field (LPF) adjacent to the final margin were recorded. Final margin groups were positive (carcinoma at margin), near (< or = 1/2 LPF of margin), and negative (> 1/2 LPF). There were 39 ipsilateral breast failures (IBFs), and 63 patients (10.8%) developed distant metastases (DMs). Decreasing margin distance and increasing amounts of carcinoma near the margin were associated with IBF and DM. The 5 tiered composite factor of margin distance and amount of carcinoma near margin (negative margins, near least amount, near intermediate amount, near greatest amount, and positive margins) resulted in 12-year IBF rates of 9%, 6%, 18%, 24%, and 30%, respectively (P < .001). The composite factor, margin amount of carcinoma near margin, and invasive carcinoma/initial excision specimen dimension ratio were the most precise parameters assessing excision adequacy and, ultimately, IBF risk. The amount of carcinoma near the margin and volume of excised parenchyma related directly to the amount of residual carcinoma in the adjacent breast parenchyma. PMID- 14560563 TI - Intraoperative consultation for renal lesions. Implications and diagnostic pitfalls in 324 cases. AB - Intraoperative consultation rarely is requested for lesions in the kidney. Of 324 renal lesions submitted for gross or frozen section intraoperative consultation, 199 specimens were submitted for gross consultation only; no diagnostic pitfalls were noted. The clinical implications and diagnostic pitfalls in 125 specimens submitted for frozen section were the focus of our study. Frozen section intraoperative consultation was requested to evaluate surgical margins in partial nephrectomy specimens, solid renal mass in an unusual clinical or radiologic setting, synchronous renal and extrarenal masses, cystic renal lesion, ureteral surgical margins for transitional cell carcinoma, multiple renal masses, solid mass in a diffusely cystic kidney, and renal injury. Among the 125 cases, the diagnoses were deferred in 17 (6 renal cell carcinomas with granular cytoplasm, 7 cystic lesions, 3 metastatic tumors, 1 leiomyoma). The frozen section diagnoses were incorrect owing to limited sampling in 5 and misinterpretation in 4 (melanoma vs angiomyolipoma, lymphoma vs angiomyolipoma, benign cyst vs cystic renal cell carcinoma, metastatic renal cell carcinoma vs pheochromocytoma). Awareness of distinctive indications for frozen section intraoperative consultation and diagnostic pitfalls should improve diagnostic accuracy and facilitate proper management of these lesions. PMID- 14560564 TI - Detection of hepatitis C virus RNA in formalin-fixed, paraffin-embedded thin needle liver biopsy specimens. AB - We examined the feasibility of detecting hepatitis C virus (HCV) RNA in formalin fixed, paraffin-embedded (FFPE) specimens obtained by thin-needle biopsy (TNB). Specimens obtained by large-needle biopsy (LNB) and unfixed frozen tissue served as controls. A total of 23 biopsy specimens, 13 obtained by TNB and 10 by LNB, from 20 patients with chronic hepatitis C were included in the study. HCV RNA was detected by nested reverse transcription-polymerase chain reaction (RT-PCR). HCV RNA was found in FFPE archival specimens obtained by TNB (11 [85%]) or LNB (7 [70%]). The sensitivity was similar in unfixed tissue in which HCV RNA was found in 7 (88%) of 8 TNB specimens and 6 (86%) of 7 LNB specimens. The detection of HCV RNA did not seem to be affected by storage of the paraffin blocks, the presence of advanced fibrosis and cirrhosis, or fragmentation of the core cylinder. TNB yields enough qualitative suitable material to detect HCV RNA by RT PCR. PMID- 14560565 TI - Expression of human mucin genes during normal and abnormal renal development. AB - Human mucin genes encode large O-glycoproteins, which are expressed in various epithelial tissues. The proteins are the main components of mucus, but also might be involved in morphogenesis of or carcinogenesis in many organs. We studied the expression of human mucin genes during fetal kidney development and in malformed cystic renal diseases in 10 normal fetal kidneys and 12 malformed kidneys by in situ hybridization and immunohistochemical analysis. MUC1, MUC3, and MUC6 were expressed in normal fetal kidney. MUC1 was expressed from 7.5 weeks of gestation in the metanephric blastema and throughout fetal life in the ureteric buds, distal convoluted tubules, and collecting ducts. MUC3 was expressed weakly in immature tubules from 8 weeks of gestation, after which it was expressed weakly and focally in the proximal convoluted tubules. MUC6 was expressed at 9.5 weeks of gestation in the tips of the ureteric buds and later in the collecting ducts. In malformative cystic diseases, only MUC1 expression was retained; no expression of MUC6 and MUC3 was observed. These results implicate human mucin genes (MUC1, MUC3, and MUC6) in renal morphogenesis processes. PMID- 14560566 TI - Babesiosis. Two atypical cases from Minnesota and a review. AB - We present 2 atypical cases of babesiosis and a review of babesiosis. The first patient was a 72-year-old man with an intact spleen, who had marked intravascular hemolysis. His RBCs were parasitized heavily with trophozoites of Babesia, and he had a large number of extracellular aggregates of Babesia. The infection did not respond to oral antibiotic therapy, and he required an RBC exchange transfusion. The second patient was a 29-year-old man who had undergone splenectomy and who had multiple episodes of fever and gastrointestinal symptoms for 4 months, with partial response to antibiotics. Thin smears revealed both intraerythrocytic and extraerythrocytic forms in very low numbers. The infection responded promptly to clindamycin and quinine therapy. The varying clinical manifestations, from acute to chronic, at a wide range of ages and often the difficulty of detection by routine blood smears make it necessary that a high index of clinical suspicion be present for prompt diagnosis. With increasing numbers of cases of transfusion transmitted babesiosis being reported, protection of the blood supply is essential. PMID- 14560567 TI - A high proportion of novel mycobacteria species identified by 16S rDNA analysis among slowly growing AccuProbe-negative strains in a clinical setting. AB - Sequencing of the 16S ribosomal DNA (rDNA) for identification of nontuberculous mycobacteria (NTM) has contributed to the establishment of more than 35 new species during the last decade. Increasingly, NTM are accepted as potential or proven pathogens. We identified, by 16S rDNA sequence analysis, slowly growing NTM isolates negative by AccuProbe (GenProbe, San Diego, CA) that previously were identified by using conventional biochemical techniques, to determine the accuracy of reporting AccuProbe-negative NTM prior to sequence-based identification. Of 82 strains, 30 were deemed novel. An attempt was made to determine the clinical importance of previously misidentified novel species. Clinical cases are described for a number of strains previously identified as Mycobacterium terrae complex, Mycobacterium scrofulaceum, and Mycobacterium avium complex. As sequence-based identification methods become more commonplace in clinical microbiology laboratories, there is a need to understand the significance of previously undescribed species, which often mimic and subsequently are identified as well-established species. PMID- 14560568 TI - Immunostaining of visceral leishmaniasis caused by Leishmania infantum using monoclonal antibody (19-11) to the Leishmania homologue of receptors for activated C-kinase. AB - It sometimes is difficult to diagnose leishmaniasis in tissue sections or in smears, particularly in unusual sites or if few parasites are present in the lesion. Leishmania species must be differentiated morphologically from a variety of other microorganisms, including Toxoplasma gondii, Histoplasma capsulatum, Trypanosoma cruzi, and Penicillium marneffei. We tested the value of monoclonal antibody p19-11 raised against the Leishmania homologue of receptors for activated C-kinase (LACK) as an immunohistochemical marker for amastigotes of Leishmania infantum. We evaluated a total of 117 paraffin-embedded lesions due to L infantum (92 cases), T gondii (15 cases), H capsulatum (5 cases), T cruzi (3 cases), and P marneffei (2 cases). Amastigotes of Leishmania species were detected in 92 (100%) of the leishmaniasis lesions. There were no false-positive LACK immunoreactions in any of the toxoplasmosis, histoplasmosis, trypanosomiasis, or penicilliosis specimens (0/25). We found the anti-LACK antibody p19-11 to be a highly specific and sensitive paraffin-reactive immunohistochemical marker for the confirmation or identification of Leishmania species in tissue sections. PMID- 14560569 TI - Fatal disseminated adenovirus infections in immunocompromised patients. AB - Adenovirus has emerged as an important pathogen in immunocompromised patients, in whom disseminated disease occurs frequently and is associated with a high mortality rate. In a retrospective review of 1,847 consecutive autopsies, we identified 84 cases where adenovirus infection was suspected clinically. Adenovirus infection was confirmed at autopsy in 8 (10%) of 84 cases; all were immunocompromised patients. Six (75%) of these cases had disseminated adenovirus infection that contributed to death. Pathologic findings attributed to adenovirus infection included pneumonia with or without intra-alveolar hemorrhage, hepatic necrosis, enterocolitis with or without mucosal hemorrhage, epicardial hemorrhage, and ulcerations of the larynx, trachea, and ileum. This work shows that severe and fatal adenovirus infections are not infrequent, particularly in the immunocompromised population. Both clinicians and pathologists must become aware of the pathogenicity of adenovirus in this patient population, including its potential for causing life-threatening hemorrhage. PMID- 14560570 TI - Recombinant protein to analyze autoantibodies to proteinase 3 in systemic vasculitis. AB - The presence of antineutrophil cytoplasmic autoantibodies with specificity for proteinase 3 (PR3-ANCA) usually is detected by enzyme-linked immunosorbent assay (ELISA) with purified PR3 as a substrate. We studied the technical performance of direct and capture ELISA using a recombinant proteolytically inactive form of PR3 produced in the baculovirus expression system for the detection of PR3-ANCA in 114 patients with systemic vasculitis at diagnosis. We found that ELISA using recombinant PR3 produced in insect cells is a promising alternative for ELISA with native PR3. We found a correlation between tests using recombinant or native PR3, as well as correlation of the ELISA results with ANCA titers measured by the indirect immunofluorescence technique. However, the specificity for ANCA associated vasculitis of ELISA with recombinant PR3 was lower than ELISA using native PR3. Compared with the direct assay, capture ELISA is a more sensitive method for PR3-ANCA detection, with both native and recombinant PR3, and its results depend on the monoclonal antibody used to capture the antigen. PMID- 14560571 TI - The assessment of anti-endothelial cell antibodies in scleroderma-associated pulmonary fibrosis. A study of indirect immunofluorescent and western blot analysis in 49 patients with scleroderma. AB - We recently reported on the use of an indirect immunofluorescent method designated the rodent lung assay; this test assesses for the presence of circulating antibodies directed at components of the microvasculature. Serum samples from 49 patients with scleroderma were incubated with rodent lung tissue sections and visualized with fluoresceinated human anti-IgG. The assay also was performed on samples from a control group. Western blot analysis was performed with endothelial cell protein extracts using serum samples from patients with scleroderma and from healthy control subjects. The control subjects had a negative indirect immunofluorescent assay result. In the patients with scleroderma, there was a significant positive correlation between intensity of indirect immunofluorescent staining and pulmonary fibrosis (r = 0.316; P = .0347) and hypertension (r = 0.310; P = .0408). Western blot analysis revealed antibody binding to proteins in extracts of human endothelial cells in all patients in whom there was evidence of pulmonary disease. The indirect immunofluorescent rodent lung assay and Western blot data support a potential role of anti endothelial cell antibodies in the propagation of scleroderma-associated pulmonary disease. PMID- 14560572 TI - Bone marrow core biopsy specimens in AL (primary) amyloidosis. A morphologic and immunohistochemical study of 100 cases. AB - We retrospectively reviewed 100 bone marrow core biopsy specimens from patients with AL (primary) amyloidosis. The morphologic and immunohistochemical features were assessed by standard histochemical stains (H&E, periodic acid-Schiff, Congo red) and immunohistochemical stains for light chain immunoglobulins. Bone marrow core biopsy revealed a plasma cell dyscrasia in 83% (lambda, 65; kappa, 18) of cases. Amyloid deposits were observed in 60% of the bone marrow core biopsy specimens and, when present, were detected most often in blood vessel walls only (39/60). However, if present, interstitial amyloid deposition was significantly more associated with patients with a monoclonal kappa light chain gammopathy (P = .04). Through the careful analysis of standard histochemical and immunohistochemical stains, bone marrow core biopsy provides essential diagnostic information in cases of AL amyloidosis. PMID- 14560573 TI - ALK-positive anaplastic large cell lymphoma with leukemic peripheral blood involvement is a clinicopathologic entity with an unfavorable prognosis. Report of three cases and review of the literature. AB - Leukemic peripheral blood involvement in anaplastic large cell lymphoma (ALCL) is uncommon. We describe 3 children with such manifestations and review the features of 9 pediatric and adult patients previously described in the literature. Leukemic involvement in ALCL may occur at the time of initial diagnosis or develop during the course of disease. It most often is associated with the small cell histologic features and the t(2;5)(p23;q35). Clinical features commonly include significant respiratory distress, diffuse lung infiltrates or pleural effusions, and hepatosplenomegaly. Most cases have an aberrant T-cell immunophenotype with frequent expression of myeloid antigens, most often CD11b or CD13. Ten of the 12 cases reviewed had a poor response to therapy or early relapse. Thus, while anaplastic lymphoma kinase-positive ALCL and young patient age generally are associated with a favorable prognosis, leukemic involvement seems to identify a high-risk malignant neoplasm that requires more aggressive therapy, including hematopoietic stem cell transplantation. PMID- 14560574 TI - T-cell large granular lymphocyte leukemia of donor origin after allogeneic bone marrow transplantation. AB - A 39-year-old man with chronic myeloid leukemia in accelerated phase underwent allogeneic bone marrow transplantation (BMT). At 6 months after BMT, lymphocytosis (WBC count, 23,100/microL [23.1 x 10(9)/L]; 80% (0.80) large granular lymphocytes [LGLs]) occurred. The LGLs were CD3+CD4-CD8+, with clonally rearranged T-cell receptor gamma gene, and of donor origin, as shown by analysis of polymorphic microsatellite markers. Epstein-Barr virus was not present. The diagnosis, therefore, was consistent with T-cell large granular lymphocytic (T LGL) leukemia. Corticosteroids controlled the LGL count, but progressive pancytopenia led to death 4 months later. Retrospective analysis showed that the T-LGL leukemia apparently had arisen as early as 3 months after BMT. The distinguishing features of this case included donor origin, neoplastic nature, and the aggressive fatal outcome. PMID- 14560575 TI - Hyperplastic-like colon polyps preceding microsatellite-unstable adenocarcinomas. PMID- 14560576 TI - Careful: the health insurer near you may be a scam. PMID- 14560577 TI - Building a better balance sheet. PMID- 14560578 TI - Expenditures for prescription drugs: too much or not enough? AB - With insurance, consumers generally do not have a financial incentive to reduce or contain prescription drug use. Drug spending can be controlled by: Understanding the sources of drug cost Improving use of drug formularies Using clinical pharmacists to improve prescribing behavior. PMID- 14560579 TI - District court affirms CMS's hold-harmless policy for DSH. AB - Under CMS's hold-harmless policy, a hospital's Medicaid proxy for periods prior to 2000 may include certain state-only program days for patients who were not eligible for medical assistance under an approved Medicaid state plan if the hospital either received DSH payments that included the same type of days in previous cost-reporting periods settled before October 15, 1999, or filed an appeal citing exclusion of these days from the Medicare DSH formula before October 15, 1999. PMID- 14560580 TI - Capacity planning seeing the forest for the trees. AB - Strategies for responding to today's rise in demand for inpatient services are many. The following are a few "tried-and-true" strategies: Build new and replacement hospitals Spin off specialty services Convert space occupied by less profitable specialty services Improve patient management Reclaim mothballed space Implement urgent care tracks for appropriate ED patients. PMID- 14560581 TI - Capital allocation. Three cases of financing the future. AB - In hospitals and health systems, numerous worthwhile capital projects are competing for limited capital. Hospitals face demands for technology upgrades, facility renovations, and new services. Some projects cannot be delayed, even though they do not bring a direct financial return. Certain services are not themselves profitable but serve as patient gateways to more profitable services. PMID- 14560582 TI - Evidence-based financial management. AB - Like the practice of evidence-based medicine, evidence-based financial management can be used by providers to improve results. The concept provides a framework that managers and researchers can use to help direct efforts in gathering and using evidence to support management decisions in health care. PMID- 14560583 TI - Assessing new technology: how are other hospitals facing the challenge? AB - A VHA West Coast survey of 19 hospitals shows several key components aid successful new technology assessment programs, including: A multidisciplinary committee structure that includes substantial clinical representation Set criteria and protocols for technology evaluation Strong integration with the hospital's strategic plan. PMID- 14560584 TI - Rx for low cash yields. AB - Certain strategies can offer not-for-profit hospitals potentially greater investment yields while maintaining stability and principal safety. Treasury inflation-indexed securities can offer good returns, low volatility, and inflation protection. "Enhanced cash" strategies offer liquidity and help to preserve capital. Stable value "wrappers" allow hospitals to pursue higher yielding fixed-income securities without an increase in volatility. PMID- 14560585 TI - The benevolent autocrat: is it the right fit for the times? PMID- 14560586 TI - 7 strategies for increasing delivery of IT value. PMID- 14560588 TI - Disturbance of timing and selfhood in schizophrenia. PMID- 14560587 TI - [History and overview on psychiatry and psychiatric care in Japan--for the new development of the Japanese Society of Psychoneurology in the 21st century]. PMID- 14560590 TI - The character of "have" in the melancholic time. PMID- 14560589 TI - Schizophrenia as a temporal mode of being: an existential "ante-festum" impatience. AB - We suggest in a phenomenological perspective to consider schizophrenia as a special form of human temporality. From this perspective, we view the symptoms of schizophrenia as actions undertaken by subjects to stabilize themselves in existence. From this vantage, we describe the clinical expression of the disorder as a type of "existential impatience", characterized by a painful and elusive "now". This present time posits the prime moment of the constitution of the person. Existential impatience reflects from our patients the persistence of excessive efforts towards individuation. Schizophrenia. In human life in general, individuation consists in an unceasing dynamic process of building up of the self. This process starts with the non-self and particularly with the other. Therefore, the emergence of any relation within the self is grounded in the relation with the other and is based on the relation the other establishes with himself. Schizophrenia distinctly displays the two constitutive moments of "being oneself." These moments are generally linked for all of us: an "unending coming to oneself" (difference of identity), and a "continuous maintenance of being a self" (identity of difference). Existential impatience is not only an irritability of a formal order. Existence itself is impatient in the schizophrenic experience as it hastens to reach human goals while trampling on an "ante-festum" temporal mode. This "before-the-feast" temporal structure is dominated by the shiver before an unknown future, a sign of a basic quest for a task. Schizophrenic "ante-festum" is both a constant fear of being unable to come to oneself and a desperate effort to reach this unknown future. If psychopathology claims to settle [establish] that "order" and "measure" would constitute the two fundamental anthropological bases of human being, impatience of existence draws the emblematic figure of the disorder of measure as a referential motion of the birth of any temporalisation. Such considerations suggest the value, in treatment and rehabilitation, of praising patience and focusing on building, or re-building, the past. The main objective is to reach a maieutics of the self based on relationships in the community and with care givers, all within an accompanying structured, daily framework. PMID- 14560591 TI - Psychoanalysis--a phenomenology of language. AB - It is often said that there is an unbridgeable gap between phenomenology and psychoanalysis. But this is only half the truth. Starting from Merleau-Ponty's proposal of an objectivistic and an idealistic divergence of Freud's enterprise the paper tries to reconstruct two possible movements: the objectivistic divergence of Husserl's enterprise as a phenomenological shift towards psychoanalysis and the idealistic divergence of Freud's enterprise as a psychoanalytical shift towards phenomenology. It is shown that this approach is possible on the field of language and semiotics where psychic life, as the essence of our subjectivity, may be rendered as a uniform phenomenon of an articulation in the world of signs. This thesis is elucidated at the end of the paper by discussing first (to the phenomenological side) a certain interpretation of the use of Husserl's reduction as a method of investigation in psychiatry and second (to the psychoanalytic side) the importance of Lacan's difference between repression and foreclosure for a psychopathological understanding of psychosis. PMID- 14560592 TI - Genetic phenomenology of transference psychosis--from the psychoanalysis of a case of "loss of natural self-evidence". PMID- 14560593 TI - [Psychiatric symptoms related to interferon treatment for chronic hepatitis]. AB - Psychiatric symptoms related to interferon (IFN) treatment for chronic hepatitis have been a crucial issue in consultation liaison psychiatry. In this report we present a hypothesis regarding the development of psychiatric symptoms. There were marked differences in the incidence of psychiatric symptoms among studies. This may be because psychiatric symptoms are readily overlooked in routine practice in the Department of Internal Medicine, and because IFN treatment frequently causes transient depression. It was speculated that psychiatric intervention was required in 10-19% of the patients. We found that among psychiatric symptoms related to IFN treatment, depression with irritation and anxiety was commonly observed. In many case reports, mood disorder was noted as a precursory or residual symptom of delirium and hallucination/delusion. We present management guidelines for psychiatric symptoms in IFN treatment, and propose that self-assessment scaling should be performed before and during treatment to detect psychiatric symptoms in the early stage, and that patients with suspected symptoms should be promptly referred to the Department of Psychiatry. We introduce an open study of antidepressant treatment for depression related to IFN treatment, and recommend aggressive administration of antidepressants. IFN treatment should be discontinued in patients with: moderate or severe suicidal ideation or suicide attempt, depression that does not respond to antidepressant treatment, manic state, hallucination/delusion, or delirium. PMID- 14560594 TI - [Music therapy used on a patient in the terminal stage of cancer who narrated a tale based on her fantasy]. AB - Music therapy was used on a patient in the terminal stage of cancer who described a fantastic tale built around her imagined pregnancy. We believe that psychiatric intervention was successful through the introduction of music therapy as part of her palliative treatment. Particularly notable in this case was that the patient dealt with the development of ascites by telling a fantastic tale related to a pregnancy--i.e., becoming pregnant with the therapist's child. Her tale started by statements such as "I am pregnant and suffering from morning sickness" and "I am in the third month of pregnancy with the child of my therapist." This progressed in parallel with the actual changes in her physiological symptoms. Among the attending physician, the patient, and her family, this imaginary tale was an important means of communication. The theme of pregnancy, love, and family developed from an unconscious dynamic process between the physician and patient, which could be interpreted as a family-related story originating in the mind of a terminally ill patient. Through this family romance with a major theme of pregnancy, the patient was able to sense her physical crises. This process may be thought of as a product of the so-called "mythgenerating capacity" under limited conditions, which was proposed by Ellenberger. Being aware of this situation, the attending physician and others in charge of her care did not insist on negating her story: instead they accepted her fantasy, and through it they succeeded in establishing psychiatric communications. The process may be considered a form of the narrative approach that is currently attracting attention. In this example, the "process of mourning" and "stages of accepting death" in the terminal state found an outlet in the creation of a fantasy. We believe that this case illustrates the important role of communication between the physician and a patient through the acceptance by the former of a fantastic tale given by the latter; and of a psychotherapeutic intervention, i.e., a sincere interaction between them through music therapy. PMID- 14560595 TI - [On criminal procedure and access to treatment for mentally disordered offenders: a pre-trial brief evaluation and it's outcome]. PMID- 14560596 TI - [Overview on the 21st century study on endocrine metabolism: from prevention to cutting-edge medical care]. PMID- 14560597 TI - [Alcoholic liver diseases: their etiological mechanism and clinical features]. PMID- 14560598 TI - [Physiopathology and treatment of systemic lupus erythematosus--changes in its presentation and future problems]. PMID- 14560599 TI - [Diabetic nephropathy--progress in its preclinical and clinical studies]. PMID- 14560600 TI - [Characteristics of legionellosis in Japan and its worldwide trends]. PMID- 14560601 TI - [Physiopathology and treatment of HAM]. PMID- 14560602 TI - [Molecular mechanism involved in estrogen action and its role in bone metabolism and brain function]. PMID- 14560603 TI - [Menopause in women and cardiovascular diseases]. PMID- 14560604 TI - [Androgen deficiency in men and resultant diseases]. PMID- 14560605 TI - [Clinical significance of dehydroepiandrosterone (DHEA) supplement therapy]. PMID- 14560606 TI - [Apoptosis-suppressing action of dehydroepiandrosterone (DHEA) and its role in longevity]. PMID- 14560607 TI - [Japanese and American guidelines for atrial fibrillation--difference in the standards for selecting anti-arrhythmic agents in the two countries]. PMID- 14560608 TI - [Basic policy in treatment in the field of internal medicine--maintenance of the sinus rhythm or rate control?]. PMID- 14560609 TI - [Goal in the treatment of paroxysmal atrial fibrillation--attack frequency, duration, or QOL?]. PMID- 14560610 TI - [Ablation therapy of atrial fibrillation--its indication and problems]. PMID- 14560611 TI - [Guidelines for anti-platelet and anti-coagulation therapies--based on evidences in the United States and Japan]. PMID- 14560612 TI - [Centennial Memorial Lecture. Hakaru Hashimoto]. PMID- 14560613 TI - [Centennial Memorial Lecture. Fighting spirit of Shibasaburo Kitasato]. PMID- 14560614 TI - [Regeneration medicine by cell transplantation]. PMID- 14560615 TI - [Cell transplantation therapy targeting at angiogenesis]. PMID- 14560616 TI - [Overview on regenerative medicine using HGF]. PMID- 14560617 TI - [Clinical practice of transplantation of regenerated blood vessels using bone marrow cells]. PMID- 14560618 TI - [Treatment of osteo-articular diseases using cultured autologous mesenchymal cells]. PMID- 14560619 TI - [Regeneration and formation of physiopathological states by cells derived from the bone marrow]. PMID- 14560620 TI - [New development in hematopoietic stem cell transplantation]. PMID- 14560621 TI - [Neurological symptoms as the initial signs of malignant tumors: paraneoplastic neuropathies]. PMID- 14560622 TI - [Clinical aspect of hepatocellular carcinoma]. PMID- 14560623 TI - [Approach to arteriosclerosis of patients with diabetes mellitus]. PMID- 14560624 TI - [Digestion and absorption--current topics]. PMID- 14560625 TI - [Clinical aspect of rapidly progressing nephritis]. PMID- 14560626 TI - [Sleep apnea syndrome]. PMID- 14560627 TI - [Clinical aspect of lymphatic tumors]. PMID- 14560628 TI - [Prion disease]. PMID- 14560629 TI - [Diagnosis and treatment of polymyositis and dermatomyositis]. PMID- 14560631 TI - [Recent findings on hypopituitarism]. PMID- 14560630 TI - [Current status and problems of coronary interventional therapy]. PMID- 14560632 TI - [Granulomatous respiratory tract diseases--with special reference to sarcoidosis]. PMID- 14560633 TI - [Physiopathology and treatment of heart failure due to poor dilatation]. PMID- 14560634 TI - [The "forty-nine" and "seventeen" problems]. AB - The single most common age of murderers in Japan is 49. Individuals aged between 45 and 54 are, however, likely to commit not offences but suicides comparing with other generations. One factor behind this phenomenon is the trend toward company downsizing, which is thought to affect the mental attitude of workers. This trend is destroying the Japanese lifetime employment system and undermining worker's loyalty to the company. On the other hand, in the broader category of felony crimes (assault, burglary, arson, murder), offenders aged 17 are the most common in Japan. This is thought to be partly due to the effect of so-called "school collapse" (school system failure) on the mental attitude of students. "School collapse" implies dysfunction of educational practice as a result of chattering, bullying and violence in the classroom. It is the reflection of students' dissatisfaction with the group-oriented school system. Therefore, students are unable to find any purposes in school and exhibit aggressive behaviours. In terms of mental health for both middle-age and teenage generations, it is necessary to devise new strategies whereby people can break away from the pressure of loyalty to the group. For both "normal" and "disabled" workers, rights of recuperations, adequate unemployment benefits policy, and support system for occupational change should be established. Simultaneously, for both "normal" and "disabled" students, reform of the school system under the concepts of diversity of education, rights of selection, and rights of refusal is urgently necessary. With respect to forensic psychiatry, without the provision of social supports, labeling of conduct disorder as well as other disorders should be avoided. In particular, instead of introducing security hospitals, drastic improvements of medical reformatories and medical prisons are necessary to provide adequate medical and social supports for offenders with psychiatric disorders. PMID- 14560635 TI - [Legal welfare for lawbreakers with mental retardation]. AB - In Japan, most lawbreakers with psychiatric disorders are committed to hospital by order of the relevant prefectural governor under the Mental Health Act. This can be called the "compulsory treatment policy" for psychiatric patients. On the other hand, most law-breakers with mental retardation (except those with severe and profound retardation) are prosecuted and sentenced to imprisonment under the Criminal Law. This can be called the "strictly judicial policy" for mildly or moderately retarded people. In my opinion, instead of either compulsory treatment or strict justice, "compassionate justice", or welfare support in the field of justice, is crucial to protect individuals with psychiatric disorders and/or mental retardation from the stigma of criminality. PMID- 14560636 TI - The diversity and challenge of conduct disorder. PMID- 14560637 TI - [Yamatogawa scandal denounces psychiatric treatment system of Japan]. AB - OSAKA Center for Mental Health and Human Rights, established in 1985, has been taking action on the Yamatogawa Hospital Affair. This hospital had developed many scandals for a long time. For example, the inpatients were murdered through the violence by the hospital staffs in 1969, 1979 and 1993. Despite the repeated accusations, this hospital had been allowed to operate until Oct. 1997, when Osaka Prefecture Governor ordered the hospital to stop its operation. Throughout the entire course of this Affair, the Psychiatric Review Board of Osaka Prefecture (regulated and expected to protect the patient's rights by the Mental Health Law and its amendment, the Mental Health and Welfare Law) did not function. The prohibition articles of the Law about restriction on actions, such as correspondence and interview, were not effective and had been neglected. The responsible department of the Prefecture Government had not started the valid investigation into the hospital. We demand the valid measures to protect the human rights of psychiatric patients. The author proposed the independence of the Psychiatric Review Board from the Prefecture Government, the start of the ombudsman system and so on. "Principles" by the UN (1991) should be cleared. PMID- 14560638 TI - [Unfair staff-patient ratios of mental hospitals in Japan]. AB - Japanese Medical Service Law regulates the staff-patient ratios of the hospital beds which are divided into four categories: tuberculosis, psychiatric, infectious diseases and others. The staff-patient ratios for mental hospitals have been regulated under the lowest level since 1958. Consequently, there have been frequent use of restraints and segregation of patients. A committee was set up to review the above standards in the fall of 2000 for the first time. The conclusions drawn by the committee for the change of various standards including staff-patient ratios for mental hospitals were far from desirable from the view of mental health professionals and patients' family members. At this presentation, the reasons for the difficulty of improving the staff-patient ratios in Japanese mental hospitals will be noted with critical analysis, the author having been the member of the hospital standard review committee. PMID- 14560639 TI - [Integration of psychiatric treatment and rehabilitation]. AB - Residual symptoms exist in psychiatric disorders, especially in schizophrenia. These become more serious according to the length of stay in hospital. Therefore, rehabilitation programs are necessary throughout the hospitalization. In the acute state also, psycho-social therapy and education are important. The most important is to give them the motivation toward their post-hospital life. Short stay in their residence before discharge gives us the information about the problems of patients' community lives. And to the patients and their families, it gives the confidence of community-living. 'Home-visit' by nursing staffs during this short stay further shows us how to support them and their families. These rehabilitation and support programs being prepared since 1986, the hospital-beds decreased from 603 to 505, admitted patients increased from 444 to 1586, and average length of stay shortened from 473 to 108 days. Whereas being treated outpatients increased from 2010 to 3439 for 10 years after 1990. PMID- 14560640 TI - An international perspective on Japanese mental health law: ethics and policy. AB - The challenge remains in international mental health legislation trends to find appropriate balances between specific cultural norms and appropriate international mental health standards for assessment, treatment, and release of persons suffering from mental disorders. It is important that cultural norms not be used as an excuse for inaction or substandard delivery of services. Furthermore, the international community of experts must agree upon a course of action to produce a set of guidelines which can become the point of reference for accelerating a timely delivery of improved services both in and outside hospital. Criteria for admission and release should not be compromised by short-term economic considerations nor by the local limitations imposed by the accepted corporate structures in place. It may be necessary for major re-thinking to occur, in order to link the hospital and the community in an interchange of services that will be to the benefit of patients and their families. To this end community treatment orders, intensive community treatment teams, and special follow-up procedures of care have to be applied and then monitored by responsible government agencies. After years of questionable applications of law reform in Japan it is imperative that critical self-assessment be done to give proper meaning to both the spirit and rule of mental health law in Japan. PMID- 14560641 TI - [Mental health and welfare law to build community support]. AB - While the rest of the world has experienced or completed deinstitutionalization in the turn of the century, Japan has not come up with a clear plan to decrease mental hospital beds. There are 330,000 people in hospitals, and at least 1/3 of them for social reasons. Japanese hospital-centered policy has been enrooted on paternalism that hospitals are responsible for the care of the severely mentally ill. Enactment of Mental Health and Welfare Act in 1995 was epoque making because this law is provided an opportunity for people with psychiatric disabilities to live in the community with support services. With this act, Central government has introduced care guidelines how to coordinate services by care managers in each municipalities in 2002. It is important to address that this new treatment model be based on individual's need and support given to improve an individual's quality of life in the community. PMID- 14560642 TI - [Current social workers' task: promoting a life model]. AB - The enactment of Psychiatric Social Workers Act was passed the National Diet in 1997. With this professional certification, psychiatric social workers are expected to help discharging more mental patients. For the further development of community care system, community agencies need to work as equal partners. It is critical to developing a team concept in the time that Japan is implementing case management service for the mentally ill. We should be able to work for the cause and think alike what we are for. Whether Japanese care management service empower the consumers or not depends on what model this service is based on, and whether community agencies and consumers working together. Social workers need to advocate for mental health consumers' choose from available resources, consent to treatment, create natural support networks, feel empowered, and allow themselves to live as an integral part of the community. PMID- 14560643 TI - [Contribution that Japanese social workers can make to mental health services- international evaluation of Japanese mental health and welfare services and objectives for psychiatric social workers (PSW)]. PMID- 14560645 TI - Summary of main points made at the Yokohama symposium. PMID- 14560644 TI - Developing a national strategy for mental health. PMID- 14560646 TI - [Proposal for fundamental reform of the Japanese mental health system in the 21st century. The Japanese Federation of the Associations for Mental Health Professionals]. PMID- 14560647 TI - [Requests from neuropathology to psychiatry: based on memoirs and prospect by a clinical neuropathologist]. AB - The subjects for neuropathology up to now, such as degenerative brain diseases, developmental impairments and dementia, should remain to be the one for general psychiatry in correlation with medical service and welfare. Thus, the roles of neuropsychiatry are thought to be more important as a subspeciality of general psychiatry. PMID- 14560648 TI - [A case of pregnant woman with severe obsessive-compulsive disorder successfully treated by modified-electroconvulsive therapy]. AB - Obsessive-compulsive disorder (OCD) is rare psychiatric disorder during pregnancy, and is often therapy-resistant. We report a 36-year-old pregnant woman with severe obsessive-compulsive disorder successfully treated by modified electroconvulsive therapy. During the pregnancy, severe mysophobia and compulsive washing appeared, so severely that she was unable to lie down, endangering the life of the fetus. Since the pharmacotherapy was ineffective, modified electroconvulsive therapy (m-ECT) was performed in cooperation with the obstetrician and the anesthesiologist, along with monitored cardiotocography throughout the procedure. During the second therapy late deceleration on the fetal cardiotocogram occurred, but rapid intravenous administration of ritodrine led to the cessation of abnormal uterine contraction. Two courses of m-ECT markedly diminished her symptoms, and she delivered a healthy infant without complications. M-ECT can be an alternative treatment for pregnant patients with OCD. PMID- 14560649 TI - [A study of the relationship between community networking and confidentiality]. AB - Legal and ethical conflicts arise in building a community network for mental health workers with confidentiality. Another ethical problem is encountered by involving mental health volunteers who are not obliged to be confidential. The authors made a questionnaire from the following six points of view: 1) the current attitudes of mental health professionals and workers toward confidentiality, 2) the degree of disclosure of individual information, 3) the relation between collaboration with family and confidentiality, 4) the range of extended confidence, 5) the extent of sharing individual information with volunteers, 6) the guidelines of confidentiality. The questionnaire was delivered to the following objects: 1,471 mental health professionals, 3,400 mentally disabled people and 3,400 of their family members. The returned questionnaires were analyzed and led to the following conclusions. The authors also conducted a bibliographical investigation on this subject. 1) Attitudes toward confidentiality varied among mental health professionals and also among mental health facilities. This created difficulties and confusion among them. 2) Transmission of individual information to the caregiving family members is important to help the mentally disabled, however, mental health professionals varied in the extent to which they informed them. 3) Mental health professionals in the same institute were regarded as an extent of extended confidence by 67.5% of the mentally disabled and 80.5% of family members. The caregiving family members were also regarded as such by 58.4% of the mentally disabled and 69.5% of family members. 4) The methods of transmission of individual information within a facility and/or with other facilities varied among mental health professionals. 5) A contract of confidentiality with volunteers was made only by 8.1% of mental health professionals. However, 34.1% of the mentally disabled and 24.0% of family members refused to transmit individual information to such volunteers. 6) Disclosure of individual information to the mentally disabled or their representatives is inevitable for them to authorize transmission of their individual information to others. Authorization of transmission of their individual information without knowing the content of such records is not valid authorization. 7) It is necessary to establish guidelines on confidentiality, according to 42.1% of the mentally disabled and 44.2% of their family members. PMID- 14560650 TI - [Real-time PCR quantification of bcr/abl chimera and WT1 genes in chronic myeloid leukemia]. AB - Quantification of mRNAs deriving from malignant cells is useful for estimating leukemic states. In this study, we have developed RT-PCR methods using real-time PCR detection system, a LightCycler, for quantification of bcr/abl chimerical genes in peripheral blood and bone marrow of chronic myeloid leukemia patients. Total amounts of RNA extracted were corrected using beta-actin gene as an internal standard. The coefficients of variation of intra-assay variation and inter-assay variation for each gene were within a range of 1.7-26.0% which showed more precise quantification than the competitive PCR method. The coefficients of variation of assay are within a range of 7.7-27.6% in the case of using three samples of normal subjects from blood collecting to quantification of bcr gene. Bcr/abl and WT1 genes could be measured from 10(2) to 10(8) copies and 10 to 10(5) copies with linearity, respectively. Using real-time PCR detection with LightCycler system, 2 x 10(3) K562 cells among 2 x 10(6) total cells demonstrated the bcr/abl gene, while 2 x 10(1) K562 cells among 2 x 10(6) total cells could be detected using the nested PCR method. In tests of seven clinical samples, five samples demonstrated bcr/abl and WT1 genes, while those in two other patients after bone marrow transplantation and a normal subject could not detected. This result suggests that our quantitative method reflect the clinical stages of CML patients. PMID- 14560651 TI - [Immunochemical properties of free mu-chain protein in a patient with mu-heavy chain disease]. AB - We report a case of mu-heavy chain disease. A 56-year-old woman presented with anemia and hemorrhagic diathesis. The serum of the patient was found to have free mu-heavy chain. The patient also had a kappa type-Bence Jones protein in serum and urine. Immunoelectrophoresis showed an abnormal precipitin line in the alpha 2-globulin region which reacted with antiserum to mu-chain but not with antiserum for light chains. The molecular weight of the monomer of the patient's mu-heavy chain protein was approximately 67,000 daltons less than that of the normal mu heavy chain protein. PMID- 14560652 TI - [Detection of monoclonality of gastric MALT lymphoma using PCR method and its clinicopathological application]. AB - Twenty cases of H. pylori-related gastric lymphoproliferative disorders including 12 cases of MALT lymphoma (Grade 5 after Wotherspoon, et al), 6 of suspicious of MALT lymphoma (Grade 4), and 2 of active chronic gastritis (Grade 3) were studied. Using the nested PCR method, paraffin-embedded gastric biopsy specimens of these 20 cases were investigated whether or not monoclonal IgH rearrangement could be demonstrated in infiltrating lymphoid cells. Monoclonal IgH rearrangement was recognized in 6 of 12 MALT lymphomas, but in other 14 cases including 6 of MALT lymphomas the monoclonality was not recognized. The result showed the sensitivity was 50% and the specificity was 100% with this method. Follow-up study after eradication of H. pylori was performed on 6 cases of MALT lymphoma which had showed the monoclonality before, using the same method. The monoclonality disappeared in 5 of 6 and histology showed a complete remission. In the remaining one case the monoclonality was yet demonstrated and lymphoma cells were histologically recognized. Thus, this method is very useful to assess not only the histological diagnosis of MALT lymphoma but also the effectiveness after treatment. PMID- 14560653 TI - [Antioxidant activities of green and black teas determined by the cumene hydroperoxide/hemoglobin.methylene blue method]. AB - Antioxidant activity in tea was measured by the new cumene hydroperoxide/hemoglobin.methylene blue(CHP/Hb.MB) method developed in our laboratory. Using the CHP/Hb.MB method, we investigated the activities of polyphenols(11 varieties) in order to determine their reactivity on CHP. According to the CHP/Hb.MB method, an increase in the number of hydroxyl groups in polyphenols induced high antioxidant activity. We found that this method was capable of measuring the antioxidant activity of polyphenols. Consequently, we were able to measure the antioxidant activities of heated, green, powdered and black teas by this method. The average of antioxidant activities of heated green tea was 207 nmol/ml, while that of green tea was 280 nmol/ml, powdered green tea was 481 nmol/ml and black tea was 215 nmol/ml respectively. PMID- 14560654 TI - [Influence of reagent evaporation on other laboratory reagents and appropriate countermeasures]. AB - When the automated analyzer was first introduced, abnormally high values of urine creatinine and abnormally low values of serum creatinine were seen. It was shown that evaporation of the reducing agent stabilizer(sulfurous acid), which is present in the serum iron measurement kit, was occurring and that the observed low values of serum creatinine were due to consumption of hydrogen peroxide by gasifying sulfurous acid. After the cause was elucidated, the abnormal lowering of creatinine values was prevented. This experience demonstrates that care must be exercised when introducing new analytical method in clinical laboratories because evaporation of reagents can influence in other reagent systems. PMID- 14560655 TI - [Role of laboratory medicine in medical care--from the point of view of a manager]. AB - The Japanese health insurance system has been praised because of the comprehensive coverage of medical costs. The long lifespan of Japanese, the longest in the world, however, because economic conditions are getting worse and medical costs are increasing with the increase of aged people in Japan, the health insurance system is under pressure. The Japanese government is going to start to reduce costs of medical care by applying a system like the DRG/PPS system in the USA, called Diagnosis Procedure Combination(DPC), next year in university hospitals. Under the DPC system, laboratory tests will be non-profit. We have to change the management style of clinical laboratories to cut costs for tests and labor as much as possible, choose tests selectively, respond rapidly to clinical needs, and utilize information technology efficiently for processing of laboratory data. Therefore, corresponding to changes in the medical care system, laboratory medicine has been changing. Commercial clinical laboratories are expanding their pressure in hospitals, and in-hospital laboratories are having a very hard time coping with the pressure. Since laboratory medicine is an essential part of clinical practice, we do not have to be antagonistic. PMID- 14560656 TI - [Positioning of registered clinical laboratories in proper health care]. AB - "Proper Health Care" is deemed to mean "Patient-conscious Health Care". On this basis, we have four tasks to achieve. They are: improvement in testing precision (quality control), heightened sense of medical ethics, satisfying the needs of medical institutions to outsource laboratory tests, and increased contribution to community health care. We are certainly aware that these tasks cannot be achieved overnight, however, we are determined to do our very best to reach our goal through day-to-day efforts in providing our customers with consistent and reliable services. PMID- 14560657 TI - [Future guidelines for medical technologists]. AB - In this symposium, ways in which medical technologists can contribute as supporting members in Japan's health care system will be discussed. Improvement in the job specifications of health care professionals has been identified as a major pillar of Japanese health care reform. As members of the medical support team, along with university professors and medical laboratory researchers, medical technologists will be expected to improve their knowledge and skill. Medical technologists should therefore be prepared to make great efforts to progress according to expectations, and these things must be kept in mind as a future guideline for medical technologists in Japan. PMID- 14560658 TI - [A new genotyping method, PCR-CTPP]. AB - The polymerase chain reaction with confronting two-pair primers (PCR-CTPP) is an inexpensive genotyping method, which is applicable for most single nucleotide polymorphisms(SNPs). In this method, allele-specific DNA products are amplified by applying appropriately designed two-pair primers (four primers) into a conventional PCR tube, followed by agarose gel electrophoresis. Duplex or triplex PCR-CTPP is also possible in a single tube, which reduces time and costs. An example of multiplex PCR-CTPP is described for NQO1 C609, GSTM1, and GSTT1. This convenient genotyping tool may be useful in health checkup to detect high risk individuals for some diseases, and to prevent such individuals from developing the diseases. PMID- 14560659 TI - [Urate transporter and renal hypouricemia]. AB - Urate, a purine metabolite, is a cause of gout(hyperuricemia), which is an independent risk factor for cardiovascular disease. Urate is a scavenger of reactive oxygen radicals that are involved in numerous diseases. Because humans have a renal urate reabsorption system and have lost hepatic uricase by mutational silencing in evolution, urate is present in human blood at high levels. We identified the long-hypothesized urate transporter in the human kidney (URAT1, encoded by SLC22A12), a urate anion exchanger regulating blood urate levels and targeted it with uricosuric and antiuricosuric agents. Moreover, we demonstrated that patients with renal hypouricemia have mutational defects in SLC22A12. PMID- 14560660 TI - [May-Hegglin anomaly--from genome research to clinical laboratory]. AB - The autosomal dominant macrothrombocytopenia with leukocyte inclusions, May Hegglin anomaly (MHA), Sebastian syndrome (SBS) and Fechtner syndrome (FTNS), are rare disorders characterized by a triad of giant platelets, thrombocytopenia, and characteristic Dohle body-like cytoplasmic inclusions in granulocytes. Epstein syndrome (EPS) is another autosomal dominant macrothrombocytopenia associated with Alport syndrome but without leukocyte inclusions. We previously mapped the locus for MHA on chromosome 22q12.3-q13.2 by genome-wide linkage analysis and found that MYH9, the gene for the nonmuscle myosin heavy chain-A (NMMHCA), is responsible for the disorder, by positional cloning. Mutations of MYH9 have also been found in SBS, FTNS, and EPS; the term "MYH9 disorders" has been proposed, but clear phenotype-genotype relationships were not found. We developed an immunofluorescence technique for conventional air-dried peripheral blood smears and studied the neutrophil NMMHCA localization in MYH9 disorders. Abnormal subcellular localization of NMMHCA was observed in every neutrophil from individuals with MYH9 mutations and the localization pattern was classified into three groups according to the number, size, and shape of the NMMHCA-positive granules. Patients without neutrophil inclusions on conventional May-Grunwald Giemsa-stained blood smears but with abnormal NMMHCA localization on immunofluorescence analysis had MYH9 mutations. In contrast, patients with EPS and isolated macrothrombocytopenia with normal NMMHCA localization had no MYH9 mutations. Immunofluorescence analysis of neutrophil NMMHCA is useful as a novel screening test for the differential diagnosis of macrothrombocytopenia and clear hematopathological classification of MYH9 disorders. PMID- 14560661 TI - [A new method of complete peripheral margin assessment in breast conservative surgery using an adjustable polygonal prism mould]. AB - Conventional margin evaluation for breast conservative surgery is usually based on the sections taken perpendicular to the inked margins and has difficulty in completeness. We have developed a new method using an adjustable mould during fixation so that the three-dimensional specimen is fixed in the shape of polygonal prism. The new method enables us to assess peripheral margins completely by examining the inner surfaces of the marginal slices cut parallel to the flat peripheral margins of the specimen. DESIGN: We have applied the new method to 59 invasive carcinomas and 10 noninvasive carcinomas of the breast, which were judged to be negative for residual tumor by conventional inked margin on the section cut through the center of the tumor. RESULTS: The new method detected 13(22.0%) and 3(30.0%) cases with positive margins in 59 invasive carcinomas and 10 noninvasive carcinomas of the breast, respectively. Nine of 13(69.2%) positive margins in invasive carcinomas were due to the intraductal components of the carcinomas. CONCLUSION: The polygon method is superior to the conventional inked margin method in sensitivity. Furthermore, it covered all the peripheral margins and pinpointed the positive sites. PMID- 14560662 TI - Physiological diversity and niche adaptation in marine Synechococcus. AB - During the twenty years or so since the discovery of tiny photosynthetic cells of the genus Synechococcus in marine oceanic systems, a tremendous expansion of interest has been seen in the literature pertaining to these organisms. The fact that they are ubiquitous and abundant in major oceanic regimes underlies their ecological importance as significant contributors to marine C fixation. Recent advances in the physiology and biochemistry of these organisms are presented here, focusing on strains of the MC-A and MC-B clusters; it is stressed that the data contained herein should be put into the context of the ecological niche occupied by particular genotypes in situ. This system is ripe for joining the often separate disciplines of molecular ecology and microbial physiology and provides a great opportunity to tease out the underlying processes that both mediate organism evolution and also the environmental factors that dictate this. PMID- 14560663 TI - Adoption of the transiently non-culturable state--a bacterial survival strategy? AB - Microbial culturability can be ephemeral. Cells are not merely either dead or alive but can adopt physiological states in which they appear to be (transiently) non-culturable under conditions in which they are known normally to be able to grow and divide. The reacquisition of culturability from such states is referred to as resuscitation. We here develop the idea that this "transient non culturability" is a consequence of a special survival strategy, and summarise the morphological, physiological and genetic evidence underpinning such behaviour and its adaptive significance. PMID- 14560664 TI - The biodiversity of microbial cytochromes P450. AB - The cytochrome P450 (CYP) superfamily of genes and proteins are well known for their involvement in pharmacology and toxicology, but also increasingly for their importance and diversity in microbes. The extent of diversity has only recently become apparent with the emergence of data from whole genome sequencing projects and the coming years will reveal even more information on the diversity in microbial eukaryotes. This review seeks to describe the historical development of these studies and to highlight the importance of the genes and proteins. CYPs are deeply involved in the development of strategies for deterrence and attraction as well as detoxification. As such, there is intense interest in pathways of secondary metabolism that include CYPs in oxidative tailoring of antibiotics, sometimes influencing potency as bioactive compounds. Further to this is interest in CYPs in metabolism of xenobiotics for use as carbon sources for microbial growth and as biotransformation agents or in bioremediation. CYPs are also current and potential drug targets; compounds inhibiting CYP are antifungal and anti-protozoan agents, and potentially similar compounds may be useful against some bacterial diseases such as tuberculosis. Of note is the diversity of CYP requirements within an organism, ranging from Escherichia coli that has no CYPs as in many bacteria, to Mycobacterium smegmatis that has 40 representing 1% of coding genes. The basidiomycete fungus Phanerochaete chrysosporium surprised all when it was found to contain a hundred or more CYPs. The functional genomic investigation of these orphan CYPs is a major challenge for the future. PMID- 14560665 TI - The Tat protein translocation pathway and its role in microbial physiology. AB - The Tat (twin arginine translocation) protein transport system functions to export folded protein substrates across the bacterial cytoplasmic membrane and to insert certain integral membrane proteins into that membrane. It is entirely distinct from the Sec pathway. Here, we describe our current knowledge of the molecular features of the Tat transport system. In addition, we discuss the roles that the Tat pathway plays in the bacterial cell, paying particular attention to the involvement of the Tat pathway in the biogenesis of cofactor-containing proteins, in cell wall biosynthesis and in bacterial pathogenicity. PMID- 14560666 TI - Microbial globins. AB - Globins are an ancient and diverse superfamily of proteins. The globins of microorganisms were relatively ignored for many decades after their discovery by Warburg in the 1930s and rediscovery by Keilin in the 1950s. The relatively recent focus on them has been fuelled by recognition of their structural diversity and fine-tuning to fulfill (probably) discrete functions but particularly by the finding that a major role of certain globins is in protection from the stresses caused by exposure to nitric oxide (NO)--itself a molecule that has attracted intense curiosity recently. At least three classes of microbial globin are recognised, all having features of the classical globin protein fold. The first class is typified by the myoglobin-like haemprotein Vgb from the bacterium Vitreoscilla, which has attracted considerable attention because of its ability to improve growth and metabolism for biotechnological gain in a variety of host cells, even though its physiological function is not fully understood. The truncated globins are widely distributed in bacteria, microbial eukaryotes as well as plants and are characterised by being 20-40 residues shorter than Vgb. The polypeptide is folded into a two-over-two helical structure while retaining the essential features of the globin superfamily. Roles in oxygen and NO metabolism have been proposed. The third and best understood class comprises the flavohaemoglobins, which were first discovered and partly characterised in yeast. These are distinguished by the presence of an additional domain with binding sites for FAD and NAD(P)H. Widely distributed in bacteria, these proteins undoubtedly confer protection from NO and nitrosative stresses, probably by direct consumption of NO. However, a bewildering array of enzymatic capabilities and the presence of an active site in the haem pocket reminiscent of peroxidases hint at other functions. A full understanding of microbial globins promises advances in controlling the interactions of pathogenic bacteria with their animal and plant hosts, and manipulations of microbial oxygen transfer with biotechnological applications. PMID- 14560667 TI - [The superpill--solution for all vascular diseases? Hardly! Furthermore, it undermines health services and science]. PMID- 14560668 TI - [New European perspective concerning hypertension and cardiovascular risks]. PMID- 14560669 TI - [Guidelines for substitution therapy in pituitary insufficiency in adults]. PMID- 14560670 TI - [Venous thromboembolism and combined oral contraceptives. Reported adverse reactions indicate at least similar risk with the most recent contraceptive pills]. AB - Rare adverse drug reactions (ADRs) to combined oral contraceptives (COCs), such as venous thromboembolism (VTE), are seldom disclosed until a COC has been on the market for some time. Two new COCs, Yasmin and Cilest have recently been launched in several European countries with expectations of being safer than older COCs. Utilising data from spontaneous reporting of ADRs to the Medical Products Agency in Sweden and sales data, VTE incidence for Yasmin and Cilest was estimated and compared with VTE incidence for a second (Follimin) and a third generation (Desolett) COC. The reported VTE incidence for Yasmin and Cilest was higher 4.6, and 3.7 per 10,000 users per year, respectively than the corresponding VTE incidences for Follimin and Desolett of 1.9 and 2.9 per 10,000 users per year, respectively. The differences were, however, not statistically significant. We conclude that the risk of VTE associated with the most recent COCs is at least similar to that of older COCs. PMID- 14560671 TI - [A national database could solve the issue of sudden sensorineural hearing loss]. AB - A national database is being organised for the disease Sudden Sensorineural Hearing-loss, SSH (often called "Sudden deafness"). The cause of SSH is unknown. Many theories have given rise to many treatment regimens: the vascular theory, assuming a reduced blood flow to the inner ear, the infection theory, assuming bacteria or virus being the etiological agent, and the auto-immune theory, where also blood vessels may be involved. Still another theory concerns ruptured membranes, either towards the middle ear, leaking perilymph, or intra-cochlear ruptures, resulting in perilymph and endolymph mixing and giving rise to the hearing loss. Many cases recover spontaneously: up to one third completely and another one third significantly improved. The most common therapy today is high dose corticosteroids. Collecting data on a large number of patients in a database will allow the assessment of different background factors and the possible effects of therapy. We hope to obtain data on 400 patients per year during a 4 year period in this multi-centre project. A controlled study is planned to assess the effect of cortico-steroid treatment. PMID- 14560672 TI - [How to make money? You are a county council's chief physician]. PMID- 14560673 TI - [The dangerous patient--three causes of shortages within psychiatry]. PMID- 14560675 TI - [Limit physicians' right to put the patients on sick leave!]. PMID- 14560674 TI - [Increasing number of teenage pregnancies--multicultural approach in the preventive work is needed]. PMID- 14560676 TI - [Roaring cry--"citizen-salary" for all!]. PMID- 14560677 TI - [Influence of food quality and quantity on children's teeth]. AB - Sugar and tooth decays are bound closely. The mechanisms of destruction of the tooth from the glycolyse are known. The role played by the food in the development of the caries is unique. Foods sweetened of all ways, rich in sucrose are responsible of the intervening of the carious lesions. The transformation of the fermentable sugars in lactic with the absence of hygiene and the action conjugated of the bacteria leads to tooth decays in temporary and permanent denture that evolve very quickly in the absence of treatment. Reasons for which the authors put in evidence in this survey, the influence of the food sweetened on the teeth at the children while showing the cases of caries and its complications due to the excessive consumption of fermentable hydrates of carbon. PMID- 14560678 TI - [Clinical study of vertical dimension before and after orthodontic treatment in a Class II population]. AB - The authors tried to highlight orthodontics only or mixed "orthopedics + orthodontics" repercussions about vertical dimensions while comparing the measurements before and after treatment done on teleradiographies of profile of 30 hyperdivergent and hypodivergent patients with skeletal class II. The clinical conclusions have been drawn after discussion of the results. PMID- 14560679 TI - [Conventional and 3-D cephalometry: descriptive analysis]. AB - Today, the three dimension (3D) teleradiographic different construction enable a three dimensional studies of different mandible structures. Then we can approach the result obtained with the news images processing techniques like scanner. In this paper we present the different between the measure provided by different pictures (lateralis, frontalis, axiales) and the three dimensional measure which is real. The best 3D values correlation is obtained with the frontalis. PMID- 14560680 TI - An epidemiological survey of occlusal anomalies among secondary school children in Ibadan, Nigeria. AB - To assess the prevalence of some of the occlusal anomalies in Ibadan, Nigeria, an epidemiological survey of 644 secondary school children aged 12-18 years (mean age, 14.73 +/- 1.17 S.D.) was done. The sample consisting of 341(53%) males and 303(47%) females was randomly selected. About 66% were from the middle social class while 34% belonged to the working class. Scissors bite recorded the highest prevalence (6.7%) and cross bite was next with 2.2%. No statistically significant associations were found between the occlusal anomalies and socio-economic class (p > 0.05). No statistically significant gender differences were found in relation to the anomalies (p > 0.05). The prevalences of the occlusal anomalies studied were generally lower than those among northern Nigerian children. PMID- 14560681 TI - [Replacement of deciduous incisors in children: psychological aspects]. AB - The absence of the temporary incisors could be to a genetic illness or to some multiple premature extractions. These extractions are the aftermaths of the carious lesions or some traumatisms underwent by the temporary incisors. Beyond measure the loss of the space, the premature loss some temporary incisors very often assign the relational development of the child and disturb its psychological development and the aesthetic function. Across some cases clinics, the authors show that the replacement of the temporary incisors is the therapeutic ideal solution. Indeed, the child prosthesis, replacing the temporary absent incisors, solves the psychological, aesthetic and relational problems of the child. PMID- 14560683 TI - [Localization and registration of the hinge axis in black Africans]. AB - The study of the cinematic method using "SAM" and "Quick Axis of FAG" added to mandibular condyle palpation for the hinge axis limited points, show that the Black Africans mandibular condyle rotation axis position is higher (3.5 mm) and backer (2 mm) than the Caucasians. The axial points are located to between 11 and 12 mm in front of the tragus and between 7 and 8 mm below on the perpendicular line to the furrow defining the tragus superior side to the Ectocanthus. PMID- 14560682 TI - [Pregnancy and periodontal health. Study of 133 pregnant women]. AB - The pregnancy is a modified physiological state having consequences on the buccal sphere generally and on the periodontal tissues in particular. We proceeded to the periodontal exam of 133 pregnant women; in spite of a relatively acceptable hygiene, 73.7% of them present periodontal changes. These changes concern especially the superficial periodontal. We did not observe the presence of deep pockets periodontal. It emerges of our study that neither the period of pregnancy, nor the parity have influence in the importance of the periodontal hurts. PMID- 14560684 TI - [Ischemia of the lower limbs as a symptom of acute dissection of the ascending aorta]. AB - A 27-year-old man, two 54-year old men and a 64-year-old woman presented with aspecific symptoms: acute pain and tingling in the shoulders plus paraplegia; exertional dyspnoea and tingling in both feet for the past week; increasing shortness of breath and cold, pale legs and feet for the past week; acute retrosternal pain, incontinence and paraparesis. The cause was dissection of the ascending aorta (type A). Following the operation, the legs were amputated because of ischaemia. The 27-year-old man and one of the 54-year-old men died; the other two patients made a satisfactory recovery. Acute type-A aortic dissection is a life-threatening disease that must be recognised early because it is an absolute indication for emergency surgery. The initial manifestation of type-A dissection may be very aspecific. Post-operative lower limb ischaemia, as the first symptom of this disease, is rare and usually disappears after surgery for type-A dissection. In some cases, however, ischaemia persists and alters the post-operative course dramatically. PMID- 14560685 TI - [Screening: careful considerations versus commercial medicine]. AB - People no longer seem to view their state of health in a light-hearted manner. The worried well hunger for every scrap of information they can find about their physical well-being, with the result that a veritable diagnostic industry is developing. Is the early detection of a disease always better than waiting for its signs or symptoms? It is difficult to answer this question and properly weigh up the beneficial effects against the side effects. Two main reasons are given for this. First of all, studies on the early detection of disease report the results selectively: beneficial effects are overstated and side effects are underreported. A more comprehensive evaluation method is needed. Secondly, new technological and therapeutic innovations are in competition with the existing techniques under investigation, which makes a long-term evaluation almost impossible. Moreover, this approach obscures questions fundamental to screening practices. There is a widening gulf between medical screening and traditional medicine, with the result that care is being transformed into a commercial product with a huge market. These developments in diagnostics pose new scientific and ethical questions that need to be answered. PMID- 14560686 TI - [Inhalation corticosteroids for COPD: possible less mortality]. AB - General practitioners find it difficult to accurately distinguish asthma from COPD. However, this differential diagnosis is important, as the treatment of the two entities is different. Asthma is by definition steroid responsive, whereas several long-term trials have demonstrated that in COPD, the lung function is not affected by steroid treatment. Yet it is questionable whether lung function is a good parameter for evaluating treatment effects, as there are indications that steroids affect the quality of life and the number of exacerbations. Moreover, an observational study has shown that inhaled steroids might affect mortality. Observational studies do not normally provide conclusive evidence, due to the possible inclusion of many confounding factors (such as confounding by indication). Nevertheless, this observational study indicates that it might be possible to treat COPD with steroids. A large prospective randomised trial that is currently in progress will need to confirm whether or not this is the case. PMID- 14560687 TI - [Androgens, well-being and libido in women]. AB - Women with an androgen deficiency complain about a loss of libido and a lack of well-being. It is difficult to link these symptoms to low circulating androgen levels in blood. This is because these complaints often arise during a period of life characterized by major psychosocial changes. Furthermore, coinciding oestrogen deficiency may give rise to similar symptoms. Another problem is the assessment of androgen deficiency. The androgen assay used is designed for men presenting with levels much higher than normal. Finally, concentrations in the peripheral blood are a poor reflection of intracellular androgen receptor stimulation. Patients with complaints suggestive of a relative androgen deficiency (based on the pattern of complaints and low serum androgen concentrations) should first of all be counselled for psychosocial problems. Oestrogen deficiency should subsequently be ruled out or substituted. If the complaints still persist then androgen substitution can be considered. PMID- 14560688 TI - [Guideline 'Treating asthma in children' for pediatric pulmonologists (2nd revised edition). I. Diagnosis and prevention]. AB - The case history and physical examination form the corner-stones for asthma diagnosis. Establishing the correct diagnosis may be difficult in infants and preschool children; in such cases the progression of the symptoms over time is important. Routine laboratory and radiological investigations are advised against. Allergy testing may be useful in children under the age of 4 years. Lung function investigations can be used from the age of 5 to 6 years onwards. Non invasive investigations into the degree of bronchial inflammation can be performed by measuring the fraction nitric oxide in exhaled air. House dust mite reduction is a useful measure for preventing asthma if sensitisation has been demonstrated. Breast-feeding during the first 4 to 6 months of life can be considered as a preventive measure in infants with an increased risk of developing asthma and allergy. PMID- 14560689 TI - [Guideline 'Treating asthma in children' for pediatric pulmonologists (2nd revised edition). II. Medical treatment]. AB - The second revision of the guidelines for the treatment of asthma in children is largely based on the evidence of comparative studies. Short-acting beta 2 sympathicomimetics are the medication of choice for acute exacerbations and should therefore be prescribed to each patient. Inhaled corticosteroids (ICS) are the medication of choice for maintenance treatment. Starting with a high dose of ICS which is then reduced to a lower but effective level on the basis of the complaints (step-down approach) is no longer recommended, as this strategy is not more effective than a constant dosage schedule. If asthmatic symptoms persist despite ICS maintenance treatment then 3 therapeutic options are available in the following order: doubling the ICS dose, the addition of a long-acting beta 2 sympathicomimetic, and the addition of a leukotriene receptor antagonist. PMID- 14560690 TI - [Diagnostic image (158). A woman with a painful cord below and a dimple in the breast. Mondor's disease]. AB - A 35-year-old woman had a palpable painful cord under the left breast with dimpling in the breast, due to Mondor's disease. PMID- 14560691 TI - [Determinants of hospital mortality in surgical patients aged 80 years and over]. AB - OBJECTIVE: To determine prognostic factors associated with mortality in patients aged 80 years and over who were treated at the Department of Surgery within a one year period. DESIGN: Prospective. METHOD: The following items from the complications register were investigated: degree of mobility prior to the operation and the housing circumstances, presence of comorbidity in the case history (heart disease, lung disease, diabetes mellitus, dementia, urgency of admission and operation (elective, urgent, acute)), and the surgical subspecialism (gastroenterology, traumatology, vascular surgery and general surgery). The number of postoperative complications was also examined. Statistical analyses were performed using the chi 2 test and multiple logistic regression analysis. RESULTS: A total of 179 patients were included: 53 men and 126 women, with an average age of 85 years and 86 years respectively. The overall mortality was 11%: 39% in acutely presented patients and 8% in subacute patients. None of the electively operated patients died. The following factors were significantly associated with mortality: subacute presentation (odds ratio 8.5; 95%-CI: 2.8-27), acute presentation (odds ratio 72; 95%-CI: 8-737), cardiological evaluation without further measures was associated with less mortality (odds ratio 0.13; 95%-CI: 0.02-0.85). CONCLUSION: The risk of mortality increased with the urgency of presentation. A cardiological evaluation in which the patient, without the need for further additional measures, was found to be in an optimal condition was associated with a relatively low risk of mortality. PMID- 14560692 TI - [Guideline development in the Academic Medical Center, Amsterdam, the Netherlands, 1992-2001]. AB - OBJECTIVE: To evaluate a program for the local development and implementation of evidence based guidelines in the Academic Medical Center in Amsterdam, the Netherlands. DESIGN: Retrospective evaluation study. METHOD: In the period 1992 2001, 50 guideline projects had received funding. Two questionnaires were developed to monitor aspects of development and implementation of guidelines. The first questionnaire was based on the 'Appraisal of Guidelines for Research & Evaluation' (AGREE) instrument. In total 39 projects of which the original application form as well as the final report was accessible, were scored by 3 researchers. A second questionnaire was developed on the basis of an implementation model by Grol and was filled out by 46 applicants. RESULTS: The primary motive for the development of a guideline were doubts about the effectiveness of a procedure. In 79% researchers (partly) collected their own data. The future users of the guideline were involved in an early stage of the development project in 72% of all cases. In 28% potential obstructing factors were identified. Introduction of the guideline was done through oral presentations in 42% without the use of an extensive implementation strategy. Of all subsidized projects, 24% was ongoing and 74% had been implemented successfully. CONCLUSION: In the AMC-program subsidized projects often led to successfully developed guidelines, which subsequently were introduced in practice. PMID- 14560693 TI - [Hereditary hyperferritinaemia-cataract syndrome]. AB - A 67-year-old woman and a 22-year-old man, great aunt and great nephew, were examined because of hyperferritinaemia; both had been operated during adolescence for bilateral cataract. The clinical diagnosis 'hereditary hyperferritinaemia cataract syndrome' (HHCS) was confirmed after DNA-analysis, which showed a point mutation in the L-ferritin gene on chromosome 19 (32G > A, the previously reported Pavia-1 mutation). The further supervision of the patients consisted of reassurance, providing an explanation about the background of HHCS and how it differs from HFE-gene related haemochromatosis, and informing other family members. Both patients were referred back to their general practitioners. HHCS is an autosomal dominant disorder that is characterised by elevated serum ferritin in the absence of iron overload. The dysregulation of ferritin production is caused by heterogenous mutations in the iron responsive element of the L-ferritin gene on chromosome 19, which reduce the binding of iron-regulatory proteins. This results in exaggerated L-ferritin mRNA translation, which is normally tightly controlled by the intracellular iron availability. The only relevant clinical symptom is early-onset, bilateral cataract, which is due to L-ferritin deposits in the stroma of the lens. Internists and haematologists should differentiate this syndrome from haemochromatosis in order to avoid invasive diagnostics and mistreatment. Ophthalmologists should consider this syndrome in patients with congenitial or juvenile cataract. PMID- 14560694 TI - [Consultative palliative care for nausea and vomiting in the home setting]. PMID- 14560695 TI - [Breast cancer and hormone replacement therapy around menopause]. PMID- 14560696 TI - [Undertreatment of depression; causes and recommendations]. PMID- 14560697 TI - [The effect of estrogen on cognitive processing]. AB - The so-called female sex hormone, estrogen, is best known for its crucial involvement in reproductive cycle. Recent research revealed that estrogen also plays a significant neuromodulatory and neuroprotective role. Evidence from clinical and experimental studies show that estrogen modulates specific cognitive functions such as memory and learning. There are also data suggesting its positive role for the prevention and treatment of dementia and Alzheimer's disease. The present paper presents a review of research in these fields. PMID- 14560698 TI - [Consequences of disturbed sex-hormone action in the central nervous system: behavioral, anatomical and functional changes]. AB - Experimental studies revealed that transient action of sex steroids during perinatal period is crucial for the development of male sexual behavior and sexually dimorphic brain anatomy. Meanwhile, the lack of gonadal steroids in female foetus and estrogen effects at puberty determine female behavior together with female type of anatomical brain structures and of endocrine functions. In men psychic sex consists of gender identity (self-estimation), gender role (objective estimation of sex behavior). In addition, a sexual psycho-orientation (hetero-, bi- or homosexual) has been distinguished. Although it is believed that gender depends on the socio-environmental influences such as rearing, learning and individual choice, the biological factors are considered to be most important. This concept arises from recent study on patients with gender dysphoria syndrome (transsexualism). In intersexualism, in genetic men with disturbances of sexual differentiation of external genitalia because of the lack of testoterone production or action in peripheral tissues (male pseudohermaphroditism) or in genetic women with ambiguous genitalia because of the presence and action of androgens (female pseudohermaphroditism), a discordance between the formal sex (assigned after the birth) and the psychic gender may appear. In these individuals the legal sex established according to somatic and/or genetic sex at birth may be incompatible with their actual gender identity and role. The knowledge about gender identity is necessary at the decision of eventual (!) surgical correction of sex organs in patients with ambiguous genitalia. This decision should depend not on the expected, but on the actual gender identity of the individual patient. Meantime, early bilateral gonadectomy in patients with gonadal dysgenesis and male pseudohermaphroditism is an indication for life because of the highest risk of germ cell carcinoma. PMID- 14560699 TI - [Estrogens and synaptic plasticity]. AB - Estrogens influence morphology of the brain not only in structures linked to reproductive cycle and reproductive behavior but also structure engaged in memory and cognitive functions. Estrogens stimulate synaptogenesis in pyramidal neurons of CA1 field of hippocampus. Increase in the number of spines on apical dendrites in rats occurs in the prostures phase of the cycle as well as exogenous estradiol application in ovariectomized females. The new synapses are enriched in NMDA receptor and it was found that their generation involves activation of NMDA receptors, PKA and CREB. Estradiol-induced synaptogenesis is accompanied by facilitation of LTP induction. Estradiol affects pyramidal cells of CA1 probably by inhibiting GABA-ergic interneurons. It also modulates unspecific activatory systems, which contribute significantly to neuroplasticity. PMID- 14560700 TI - [Does gender exert influence on stroke?]. AB - Stroke morbidity increases with age. That is the reason why it affects especially the middle aged and elderly. Life expectancy is longer for females than males by 10 years, that is why stroke is a major problem in women. Women die twice more frequently from stroke than men (16% vs. 8%). Stroke risk factors are basically the same in spite of gender. The most important are hypertension, diabetes, dyslipidaemia, atrial fibrillation, coronary heart disease, previous stroke, smoking, alcohol abuse, obesity and lack of physical activity. Their impact, however, is different in males and females. Women with diabetes, atrial fibrillation, myocardial infarction, obese, drinking excessive amounts of alcohol and smoking are more likely to suffer of stroke than males with the same burden. A less favourable outcome after stroke has been observed in female patients- higher mortality rates and disability. It is possible that poor prognosis is related to a drop in blood estrogen concentration after menopause. Estrogen replacement therapy has not proved to be beneficial in preventing stroke and improving outcome. There are several specific conditions: pregnancy, migraine in women associated with the occurrence of stroke. PMID- 14560701 TI - [Estrogen receptors in the brain]. AB - Estrogen plays an important role in changes taking place in the brain through the regulation of growth and differentiation of axons and dendrites, influence on plasticity, support of survival as well cognitive and behavioral functions. The classical mode of estrogen action is through the activation of its receptors, transcription factors. Two such estrogen receptors have been cloned, ER alpha i ER beta. Several isoforms of ER beta have also been characterized, which may be a significant factor in the regulation of estrogen response in the brain. Additionally, actions of estrogens through membrane or intra-cellular receptors, are suggested. The functions played by ER alpha i ER beta in the brain may be explored by studies on ER alpha or ER beta knockout mice. The results of these studies point to the role of ER alpha in reproductive functions, while ER beta seems to be important in cognitive processes. Autoradiography, immunohistochemistry, and in situ hybridization have been employed in the localization of estrogen receptors in the brain. Aside from demonstrating differences in the localization and expression levels of both receptors, these studies have proven, that both ER alpha i ER beta are able to bind estrogen. However, the pattern of expression of estrogen receptors in the brain is not static, but undergoes rapid changes in response to injury. Many studies point to a wide range of estrogen actions in the brain. For that reason, a complementary mechanism of action of estrogen is proposed, based on their interaction with various growth factors, as well as on the synergistic action of this hormone through, both, the estrogen receptors and estrogen-independent pathways. PMID- 14560702 TI - [Immunosuppressive aspects of estrogen]. AB - The evidence supports from clinical and epidemiological studies demonstrates that the prevalence of various neurological diseases changes with gender. The reason for the gender difference in these diseases is still unclear, but accumulating evidence suggests a link between gonadal hormone, such as estrogens, levels and the incidence of these diseases. Numerous reports support the idea that immunological processes contribute to the etiopathogenesis of the neurodegenerative disease, such as, Parkinson's or Alzheimer's disease; psychiatric disorders, such as schizophrenia or depression; multiple sclerosis or stroke. The neuroinflammation is regulated by numerous signal molecules, including cytokines. Estrogens have been shown to play a major role in inflammatory processes. One mechanisms by which estrogens could modulated the immune reaction is regulation of the cytokines expression. This article briefly describes different biological mechanisms underlying estrogen anti-inflammatory activity. PMID- 14560703 TI - [The clinical and laboratory diagnosis of sCJD and vCJD]. AB - Clinical diagnosis of sporadic CJD is usually confirmed by a typical EEG pattern and an increased level of protein 14-3-3 in the CSF, with specificity of 74% and 84%, respectively. However, both these tests are often negative in vCJD patients. Recently MR imaging has substantially improved the diagnosis of sCJD and vCJD. In sCJD patients hyperintense signal in T2-weighted sequences was found to be present in the neostriatum (the caudate nucleus and putamen) with specificity of 93%, while in vCJD cases signal hyperintensity was found in pulvinar thalami. The "pulvinar sign" was evidenced to be highly sensitive (79%) and specific (100%) for the diagnosis of the latter form of CJD. PMID- 14560704 TI - [The central nervous system and HIV infection]. AB - Human immunodeficiency virus (HIV) may be the cause of both primary and secondary brain diseases. In this review general features of HIV-associated neuropathology are discussed. Up to 90% of patients with AIDS have a variety of HIV-related brain diseases. Primary brain diseases including lymphocytic meningitis and HIV encephalitis are attributed directly to the effect of the virus on the brain. Secondary diseases including toxoplasmosis, cryptococcosis, primary leukoencephalopathies and lymphomas result from these patients' immunodeficiency status. PMID- 14560705 TI - [CNS changes in HIV-infected patients: magnetic resonance spectroscopy]. AB - Changes that may appear in the central nervous system in the course of AIDS either result directly from HIV infection or--as is the case with opportunistic infections and some neoplasms--develop as a secondary consequence of general immunodeficiency. Neuroimaging techniques may be most useful in the differential diagnosis of these lesions. Basic principles of HIV encephalitis and progressive multifocal leukoencephalopathy differentiation in MRI scans are discussed in the paper, and diagnostic possibilities of MR imaging in some other infections (tuberculosis, toxoplasmosis, and cryptococcosis) are outlined. Special attention is paid, on the one hand, to difficulties in the differentiation between toxoplasmosis and lymphoma, and on the other hand--to the growing diagnostic utility of MR-spectroscopy in this respect. PMID- 14560706 TI - [Pathogenetic-clinical problems of Lyme borreliosis]. AB - In this article a short review of pathogenesis and clinical manifestations of Lyme disease is presented. As regards pathogenesis, attention was paid to the mosaic protein structure of the B. burgdorfieri spirochete, particularly of outer surface proteins (Osp) that influence the clinical course and diagnosis of the disease. The presence of various atypical spirochete forms: spheroplastic L (without cell walls), cystic, and granular "blebs" may lead to a chronic form of the disease and to a low efficacy of antibiotic therapy. An important part of the pathogenesis is epithelial damage, stimulating the production of inflammatory cytokines (mainly IL-1, TNF-alpha, IFN-gamma), adhesive molecules and acute-phase proteins. Moreover, in the course of the disease not only an impairment of phagocytosis and chemotaxis was found, but also B. burgdorfieri spirochete binding by antibodies into immunological complexes that may maintain chronic inflammation. In terms of the Asbrink classification, complaints predominating in the clinical picture of an early and late stage of the disease were presented, with an emphasis on neuroborreliosis. PMID- 14560707 TI - [Neuroimaging in Lyme disease]. AB - Imaging techniques are commonly used in the diagnostics of the central nervous system disorders. Abnormalities found using MRI and MRS in neuroborreliosis patients are described. PMID- 14560708 TI - [Neuroimaging and clinical manifestations of herpes simplex encephalitis in children]. AB - Mortality rates in herpes simplex encephalitis are still high because the condition is diagnosed too late and so specific treatment is delayed. We present a retrospective study of patients with herpes encephalitis. If in a pediatric patient with CNS infection MR images show high intensity signal lesions in characteristic localization, herpes etiology should be taken into consideration, so as to undertake appropriate treatment possibly early. PMID- 14560710 TI - [Tuberculosis of the central nervous system: case reports]. AB - Tuberculosis of the central nervous system possess a serious clinical and radiological problem, since early treatment of this condition is crucial for the patient's life, and tuberculous process in the CNS may mimic many other pathologies. In this paper three cases of tuberculous process difficult to diagnose are presented. In all the cases early treatment could be started due to MRI findings suggesting tuberculous etiology. The authors emphasize the importance of presumptive radiological diagnosis in reducing the morbidity and mortality rates due to CNS tuberculosis. PMID- 14560709 TI - [Neuroimaging in a chronic demyelination process following tick-borne encephalomyelitis]. AB - We present a case of a 17-year-old female patient with a tick-borne neuroinfection. Tick-borne encephalitis is a viral disease of the CNS. Tick-borne encephalitis is usually of diphasic type, with partial epileptic seizures. No such symptoms were observed in this case. Since areas of demyelination could be seen on CT and MRI scans, immunological etiology should be taken into consideration. The correct diagnosis was established on the basis of serological examination and neuroimaging findings. PMID- 14560711 TI - [Embolic and thrombotic changes in numerous CNS vessels in the course of aspergillosis]. AB - A case of aspergillous encephalitis in a diabetic patient with acute sinusitis is presented. MRI investigation revealed characteristic changes in the brain and skull bones, including these of face and skull base. Despite a radical surgery of the sinuses and early treatment with Amphotericin B, the disease progressed rapidly and was fatal. Post-mortem neuropathological examination confirmed the diagnosis of encephalitis caused by aspergillosis. PMID- 14560712 TI - The patient's motivation in seeking complementary therapies. AB - The motivations of cancer patients in seeking complementary therapies are, fundamentally, self-healing motivations which, when engaged appropriately, can contribute to the patient's psychological and physical well being. In this paper, we apply a theoretical model, the Risk Adaptation Model, to furthering the clinical understanding of the motivations of cancer patients in seeking complementary therapies. The model identifies six discrete cognitive processes which, in combination, are hypothesized to play a central role in therapy seeking. Emphasis in this model is placed on the patient's need to maintain positive expectancies (optimism) when faced with the risk and uncertainty of cancer. This understanding of complementary-therapy seeking is grounded in the perspective that clinicians must respect the autonomy of cancer patients in their quest for appropriate therapies, and assist rather than direct their process of therapy-seeking. PMID- 14560713 TI - Smallpox revisited? AB - This article reviews the history of smallpox and ethical issues that arise with its threat as a biological weapon. Smallpox killed more people than any infectious disease in history--and perhaps three times more people in the 20th Century than were killed by all the wars of that period. Following a WHO sponsored global vaccination campaign, smallpox was officially declared eradicated in 1980. It has since been revealed that the Soviet Union, until its fall in the early 1990s, manufactured tens of tons of smallpox for military purposes. A worry is that some of this may have fallen into the hands of "rogue" nations or terrorists. Current U.S. debate questions whether smallpox vaccine should therefore be made available to the American public, which--like the rest of the world--now lacks immunity. Because the vaccine is considerably dangerous, public dialogue cannot resolve this matter if evidence material to the likelihood of attack is classified (i.e. secret). I conclude by recommending numerous future areas for ethics research related to the weaponization of smallpox. PMID- 14560714 TI - Children in clinical research: a conflict of moral values. AB - This paper examines the culture, the dynamics and the financial underpinnings that determine how medical research is being conducted on children in the United States. Children have increasingly become the subject of experiments that offer them no potential direct benefit but expose them to risks of harm and pain. A wide range of such experiments will be examined, including a lethal heartburn drug test, the experimental insertion of a pacemaker, an invasive insulin infusion experiment, and a fenfluramine "violence prediction" experiment. Emphasis, however, is given to psychoactive drug tests because of the inherent ethical and diagnostic problems involved in the absence of any objective, verifiable diagnostic tool. Effort is made to provide readers comprehensive reference sources to evidence-based reports about the serious risks these drugs pose for adults and children so that the reader may judge whether the benefits (if any) outweigh the risks for children. PMID- 14560716 TI - Prenatal diagnosis for "minor" genetic abnormalities is ethical. AB - Is it justified to detect minor genetic aberrations before birth and terminate pregnancies based upon such information? We present the case of a woman who wanted Prenatal Diagnosis (PND) to detect whether her female fetus was a Haemophilia mutation carrier. Such carriers are usually healthy. She wished to eradicate the Haemophilia mutation from her family to avoid future generations being affected and to protect her children from having to go through PND themselves. We explore existing guidelines, public attitudes and possible objections to providing PND for minor abnormalities. We argue that in a society where couples have considerable autonomy relating to decisions about the fetus at least until viability, the routine restriction of PND for minor genetic abnormalities would be an unjust infringement of individual liberty. PMID- 14560717 TI - Nazi's euthanasia files are made public. PMID- 14560718 TI - Tongue depressor. PMID- 14560719 TI - How do adult stem cells really work? PMID- 14560720 TI - Image-guided endoscopic transnasal removal of recurrent pituitary adenomas. PMID- 14560721 TI - Brain death documentation: analysis and issues. PMID- 14560722 TI - Craniocervical tuberculosis: protocol of surgical management. PMID- 14560723 TI - "Stops" in speech. PMID- 14560724 TI - Number and measure: Hermann von Helmholtz at the crossroads of mathematics, physics, and psychology. AB - In 1887 Helmholtz discussed the foundations of measurement in science as a last contribution to his philosophy of knowledge. This essay borrowed from earlier debates on the foundations of mathematics (Grassmann/Du Bois), on the possibility of quantitative psychology (Fechner/Kries, Wundt/Zeller), and on the meaning of temperature measurement (Maxwell,Mach.). Late nineteenth-century scrutinisers of the foundations of mathematics (Dedekind, Cantor, Frege, Russell) made little of Helmholtz's essay. Yet it inspired two mathematicians with an eye on physics (Poincare and Holder), and a few philosopher-physicists (Mach, Duhem,Campbell). The aim of the present paper is to situate Helmholtz's contribution in this complex array of nineteenth-century philosophies of number, quantity, and measurement. PMID- 14560725 TI - Virtual reality surgery:neurosurgery and the contemporary landscape a three dimensional interactive virtual dissection model to simulate transpetrous surgical avenues. PMID- 14560726 TI - Put into practice. PMID- 14560727 TI - Delayed ischemic deficit after resection of a large intracranial dermoid: case report and review of the literature. PMID- 14560728 TI - Galen 2003: critical analysis of brain injury in sport. PMID- 14560729 TI - Grand master of Bedlam: Roy Porter and the history of psychiatry. PMID- 14560730 TI - Tackling concussions in sports. PMID- 14560731 TI - Portraits, people and things: Richard Mead and medical identity. PMID- 14560732 TI - Neighborhood mechanisms and the spatial dynamics of birth weight. AB - This study addresses two questions about why neighborhood contexts matter for individuals via a multilevel, spatial analysis of birthweight for 101,662 live births within 342 Chicago neighborhoods. First, what are the mechanisms through which neighborhood structural composition is related to health? The results show that mechanisms related to stress and adaptation (violent crime, reciprocal exchange and participation in local voluntary associations) are the most robust neighborhood-level predictors of birth weight. Second, are contextual influences on health limited to the immediate neighborhood or do they extend to a wider geographic context? The results show that contextual effects on birth weight extend to the social environment beyond the immediate neighborhood, even after adjusting for potentially confounding covariates. These findings suggest that the theoretical understanding and empirical estimation of 'neighborhood effects' on health are bolstered by collecting data on more causally proximate social processes and by taking into account spatial interdependencies among neighborhoods. PMID- 14560733 TI - Background on the National Football League's research on concussion in professional football. PMID- 14560734 TI - The brain and nervous system in two French Renaissance "scientific" poets: Pernette du Guillet and Guillaume du Bartas. PMID- 14560735 TI - [Danish bibliographies of the history of pharmacy]. PMID- 14560736 TI - [Was Arnaldus an astrologer? Astrology in the medical thought of Arnau de Vilanova]. PMID- 14560737 TI - Ambulances on stamps. PMID- 14560739 TI - [Dealing with the French disease in early modern Augsburg]. PMID- 14560738 TI - [Statistical trends in pregnancies among adolescents in France during the past 20 years]. PMID- 14560740 TI - Life cycle. PMID- 14560741 TI - "Think tank consortium on resident work hours". PMID- 14560742 TI - The old course and the MRI. PMID- 14560743 TI - Patients' opinions matter. PMID- 14560744 TI - A transparent data repository. PMID- 14560745 TI - Unearth unflattering into? PMID- 14560746 TI - Not enough emphasis is put on patient care. PMID- 14560747 TI - The impact of a single- or double-layer closure on uterine rupture. PMID- 14560748 TI - Hormone replacement therapy and autonomic cardiovascular functions: the impact of heart rate variability analyses methods. PMID- 14560749 TI - Growth discordance in twins. PMID- 14560750 TI - Safety profile of transdermal testosterone therapy in women. PMID- 14560751 TI - Trusty resources. Reliable tools for board members. PMID- 14560752 TI - The laborist: do not repeat the mistakes of other medical systems. PMID- 14560753 TI - Gynecology knots. PMID- 14560754 TI - Medication safety issue brief. Series II, Part 4. Focusing on the human factor. PMID- 14560755 TI - Esophageal obstruction secondary to erosion of an Angelchik prosthesis: the role of endoscopic management. PMID- 14560756 TI - Intramural gastric abscess: case history and review. PMID- 14560757 TI - Diffuse nodular lymphoid hyperplasia of the colon: polyposis syndrome or normal variant? PMID- 14560758 TI - [Special issue: "Molecular mechanism of the adaptation of terrestrial plants to gravity environment on Earth"]. PMID- 14560759 TI - Gastric adenocarcinoma mimicking a submucosal tumor: case report. PMID- 14560760 TI - Helium in the treatment of respiratory failure: why not a standard? PMID- 14560761 TI - Salt saves the hot brain? PMID- 14560762 TI - Neuromuscular alterations in the critically ill patient: critical illness myopathy, critical illness neuropathy, or both? PMID- 14560763 TI - Transport of seriously ill children: a neglected global issue. PMID- 14560764 TI - [Mifepristone for induction of second trimester abortion]. AB - BACKGROUND: We wanted to assess the effect of changing the regimen for termination from gemeprost to mifepristone and misoprostol. METHOD: The study was prospective from 1994 to 2001 and included 179 women who had a second trimester termination of pregnancy because of fetal malformations. Three different regimens were used: The traditional regimen with vaginal application of gemeprost (77 patients), pretreatment with mifpristone followed by gemeprost (40 patients), and pretreatment with mifepristone followed by misoprostol (62 patients). RESULTS: The induction-to-abortion interval was significantly shorter in the two groups receiving pretreatment with mifepristone than in the group treated solely with gemeprost, on average 9.7 and 9.5 hours compared to 22.7 hours (p < 0.001). All patients who received mifepristone aborted within 24 hours of start of induction, compared to 80 % of patients treated solely with gemeprost. INTERPRETATION: The combination of mifepristone and misoprostol provides a non-invasive and effective regimen for second trimester termination of pregnancy. Pretreatment with mifepristone represents a major improvement. Misoprostol is cheaper than gemeprost, is stored in room temperature, and can be used orally as well as vaginally. PMID- 14560765 TI - Spectral cancellation of microstimulation artifact for simultaneous neural recording in situ. AB - A fundamental technical hurdle in systems neurophysiology has been to record the activity of individual neurons in situ while using microstimulation to activate inputs or outputs. Stimulation artifact at the recording electrode has largely limited the usefulness of combined stimulating and recording to using single stimulation pulses (e.g., orthodromic and antidromic activation) or to presenting brief trains of pulses to look for transient responses (e.g., paired-pulse stimulation). Using an adaptive filter, we have developed an on-line method that allows continuous extracellular isolation of individual neuron spikes during sustained experimental microstimulation. We show that the technique accurately and robustly recovers neural spikes from stimulation-corrupted records. Moreover, we demonstrate that the method should generalize to any recording situation where a stereotyped, triggered transient might obscure a neural event. PMID- 14560766 TI - Dosage individualization of erythropoietin using a profile-dependent support vector regression. AB - The external administration of recombinant human erythropoietin is the chosen treatment for those patients with secondary anemia due to chronic renal failure in periodic hemodialysis. The objective of this paper is to carry out an individualized prediction of the EPO dosage to be administered to those patients. The high cost of this medication, its side-effects and the phenomenon of potential resistance which some individuals suffer all justify the need for a model which is capable of optimizing dosage individualization. A group of 110 patients and several patient factors were used to develop the models. The support vector regressor (SVR) is benchmarked with the classical multilayer perceptron (MLP) and the Autoregressive Conditional Heteroskedasticity (ARCH) model. We introduce a priori knowledge by relaxing or tightening the epsilon-insensitive region and the penalization parameter depending on the time period of the patients' follow-up. The so-called profile-dependent SVR (PD-SVR) improves results of the standard SVR method and the MLP. We perform sensitivity analysis on the MLP and inspect the distribution of the support vectors in the input and feature spaces in order to gain knowledge about the problem. PMID- 14560767 TI - Comparison of detrended fluctuation analysis and spectral analysis for heart rate variability in sleep and sleep apnea. AB - Sleep has been regarded as a testing situation for the autonomic nervous system, because its activity is modulated by sleep stages. Sleep-related breathing disorders also influence the autonomic nervous system and can cause heart rate changes known as cyclical variation. We investigated the effect of sleep stages and sleep apnea on autonomic activity by analyzing heart rate variability (HRV). Since spectral analysis is suited for the identification of cyclical variations and detrended fluctuation analysis can analyze the scaling behavior and detect long-range correlations, we compared the results of both complementary techniques in 14 healthy subjects, 33 patients with moderate, and 31 patients with severe sleep apnea. The spectral parameters VLF, LF, HF, and LF/HF confirmed increasing parasympathetic activity from wakefulness and REM over light sleep to deep sleep, which is reduced in patients with sleep apnea. Discriminance analysis was used on a person and sleep stage basis to determine the best method for the separation of sleep stages and sleep apnea severity. Using spectral parameters 69.7% of the apnea severity assignments and 54.6% of the sleep stage assignments were correct, while using scaling analysis these numbers increased to 74.4% and 85.0%, respectively. We conclude that changes in HRV are better quantified by scaling analysis than by spectral analysis. PMID- 14560768 TI - The temporal prior in bioelectromagnetic source imaging problems. AB - The multiplicity of temporal priors proposed for regularization of the bioelectromagnetic source imaging problems [e.g., the inverse electrocardiogram (ECG) and inverse electroencephalogram (EEG) problems], is discordant with the fact that fundamental statistical principles sharply limit the choice. Thus, our objective is to derive the form of the prior consistent with the general unavailability of temporal constraints. Writing linear formulations of the inverse ECG and inverse EEG problems as H = FG + N (where the ith columns of matrices H, G, and N, are data, signal, and noise vectors at time step i, and F is the transfer matrix), and using the noninformative principle that features of the spatiotemporal prior not supplied a posteriori should be invariant under temporal transformations, we show that the implied spatiotemporal signal autocovariance matrix (of the vector formed by the entries of G) is given in block matrix form [equation in text] where Cg is a matrix of unit trace proportional to the autocovariance matrix of any column of G (representing supplied information regarding the spatial prior), epsilon[.] denotes expectation, superscript ' indicates transpose, [symbol in text] is the Kronecker product, [symbol in text] is Frobenius norm, and the "matrix scalar product" [symbol in text] indicates the inner product of the two vectors formed by the entries of the two adjacent matrices (i.e., A [symbol in text] B [triple bond] trace[A'B]). This result eliminates some uncertainties and ambiguities that have characterized spatiotemporal regularization methods--including eight methods previously introduced in this transactions. Ultimately, the result derives from an implied symmetry principle under which the form of a nontrivial noninformative temporal component of the prior can be identified. Among other things, separability of the spatiotemporal prior in terms of the above Kronecker product can be thought of as the expression of the lack of "entanglement" of the spatial and temporal contributions (a consequence of noninformativity). The approach is generalized to the important cases of non-Gaussian spatial priors, and signal and noise that are not independent (transfer matrix noise). We also demonstrate a means for computational complexity reduction, related to the application of a particular orthogonal transformation, having features dependent on whether or not the transfer matrix represents a surjective mapping. PMID- 14560769 TI - Artefact reduction with alternative cuff configurations. AB - In nerve cuff electrode recordings of neural signals, the pick-up of interfering signals can be reduced by choosing appropriate cuff configurations. In the traditionally used tripolar configuration, short circuiting of the end electrodes is expected to reduce the field inside the cuff from interfering signals. A model study suggests that moving the end electrodes toward the center of the cuff reduces the pick-up of interfering signals. In this paper, these properties are studied in more detail using a rabbit model. In addition, a new cuff configuration is suggested, which has an additional set of short circuited end electrodes. The total improvement of signal-to-noise ratio in the new configuration as compared with the traditionally used tripolar configuration was 73% for muscle signals and 127% for the stimulus pulse. PMID- 14560770 TI - Measurement of thoracic current flow in pigs for the study of defibrillation and cardioversion. AB - Although defibrillation has been in clinical use for more than 50 years, the complete current flow distribution inside the body during a defibrillation procedure has never been directly measured. This is due to the lack of appropriate imaging technology to noninvasively monitor the current flow inside the body. The current density imaging (CDI) technique, using a magnetic resonance (MR) imager, provides a new approach to this problem [Scott et al. (1991)]. CDI measures the local magnetic field generated by the current and calculates the current density by computing its curl. In this study, CDI was used to measure current density at all points within a postmortem pig torso during an electrical current application through defibrillation electrodes. Furthermore, current flow information was visualized along the chest wall and within the chest cavity using streamline analysis. As expected, some of the highest current densities were observed in the chest wall. However, current density distribution varied significantly from one region to another, possibly reflecting underlying heterogeneous tissue conductivity and anisotropy. Moreover, the current flow analysis revealed many complex and unexpected current flow patterns that have never been observed before. This study has, for the first time, noninvasively measured the volume current measurement inside the pig torso. PMID- 14560771 TI - A new method for eye location tracking. AB - A standard scleral search coil (SSC) system is supplied with a single three-axial transmitter (or with a triad of transmitters) and the direct magnetic coupling between the transmitting coils and the SSC is used for computing the eye location. The SSC's orientation components measured by the SSC system are involved in the location-tracking algorithm as well. The suggested method differs from traditional ones where the eye location is computed indirectly, relative to the measured location and orientation of a transitional three-axial magnetic sensor attached to the subject's head. The new method eliminates systematic errors caused by inaccurate situating the transitional sensor relative to the eye. It also eliminates systematic errors caused by imperfect orthogonality of the transitional sensor. It is found that an accurate location tracking with a single transmitter is possible only if orientation freedom of the eye is limited. To track the eye location with no orientation limitations, a triad of transmitters is employed. The resulting accuracy can be as good as +/- 1 mm (in a 200-Hz bandwidth) over a relatively large (0.5 x 0.5 x 0.5 m) operating region in the center of a 1 x 1 x 1 m SSC system. The analytical results obtained are confirmed with the help of computer simulations. PMID- 14560772 TI - Microwave breast imaging: 3-D forward scattering simulation. AB - Active microwave imaging (MWI) is emerging as a promising technique for the detection of biomedical anomalies such as breast cancer because of the high electrical contrasts between malignant tumors and normal tissue. Previously, we have developed fast two-dimensional forward and inverse scattering algorithms for MWI systems. In this paper, we report the full three-dimensional (3-D) forward scattering simulation in order to account for 3-D effects and to provide a fast solver in future 3-D nonlinear inverse scattering methods. The 3-D fast forward method is based on the stabilized biconjugate-gradient fast Fourier transform (BCGS-FFT) algorithm. The method has been validated for various MWI measurement scenarios. Using this fast simulation method, we demonstrate the importance of accounting for 3-D effects in MWI, and we compare numerical results with the measurements from an experimental prototype. PMID- 14560773 TI - Noninvasive imaging of cardiac transmembrane potentials within three-dimensional myocardium by means of a realistic geometry anisotropic heart model. AB - We have developed a new approach for imaging cardiac transmembrane potentials (TMPs) within the three-dimensional (3-D) myocardium by means of an anisotropic heart model. The cardiac TMP distribution is estimated from body surface electrocardiograms by minimizing objective functions of the "measured" body surface potential maps (BSPMs) and the heart-model-generated BSPMs. Computer simulation studies have been conducted to evaluate the present 3-D TMP imaging approach using pacing protocols. Simulations of single-site pacing at 24 sites throughout the ventricles, as well as dual-site pacing at 12 pairs of sites in the vicinity of atrio-ventricular ring were performed. The present simulation results show that the correlation coefficient (CC) and relative error (RE) between the "true" and inversely estimated TMP distributions were 0.9915 +/- 0.0041 and 0.1266 +/- 0.0326, for single-site pacing, and 0.9889 +/- 0.0034 and 0.1473 +/- 0.0237 for dual-site pacing, respectively, when 10 microV Gaussian white noise (GWN) was added to the BSPMs. The effects of heart and torso geometry uncertainty were also evaluated by shifting the heart position by 10 mm and altering the torso size by 10%. The CC between the "true" and inversely estimated TMP distributions was above 0.97 when these geometry uncertainties were considered. The present simulation results demonstrate the feasibility of noninvasive estimation of TMP distribution throughout the ventricles from body surface electrocardiographic measurements, and suggest that the present method may become a useful alternative in noninvasive imaging of distributed cardiac electrophysiological processes within the 3-D myocardium. PMID- 14560774 TI - A genetic segmentation of ECG signals. AB - This paper is concerned with a development of a segmentation technique for electrocardiogram (ECG) signals. Such segmentation is aimed at a lossy signal compression in which each segment can be captured by a simple geometric construct such as, e.g., a linear or quadratic function. The crux of the proposed construct lies in the determination of the optimal segments of data over which they exhibit the highest possible monotonicity (or lowest variability) of the ECG signal. In this sense, the proposed approach generalizes a fundamental and commonly encountered problem of function (data) linearization. The segments are genetically developed using a standard technique of genetic algorithms (GAs). The two fundamental GA constructs, namely a topology of a chromosome and a fitness function governing the optimization process are discussed in detail. The chromosome being coded as a series of floating point numbers contains the endpoints of the segments (segmentation points). The fitness function to be maximized quantifies a level of monotonicity of the ECG data encountered within the segments and takes into consideration differences between the extreme values (minimum and maximum) of its derivatives. As a result of the genetic optimization, we build segments of ECG signals encompassing monotonic (increasing or decreasing) regions of the signal exhibiting a minimal level of variability. A series of experiments dealing with several classes of ECG signals (namely, normal, left bundle branch block beat, and right bundle branch block beat) visualize the effectiveness of the approach and shows the specificity of the linear segments of data. Furthermore, we elaborate on the relationship between the values of the fitness function and the approximation capabilities (quantified by a sum of squared errors between the local model and the data) of the segments of the signal and show that these two descriptors are highly related. PMID- 14560775 TI - Mobile ankle and knee perturbator. AB - A mobile ankle and knee perturbator has been developed. It consists of a functional joint with an integrated clutch. Four Bowden wires connect the joint to a powerful motor and a double pneumatic cylinder. When needed during any time of the gait cycle, it is possible to impose an ankle rotation by engaging the clutch and rotating the ankle or knee joint with a predefined displacement. The system is designed to investigate electrophysiological and biomechanical features of the human ankle or knee joint during gait. PMID- 14560776 TI - The myelodysplastic/myeloproliferative disorders: the interface. AB - Most patients with myelodysplastic syndromes can be identified easily and separated from patients who have myeloproliferative disorders. A few have overlapping features, however, with evidence of morphologic dysplasia and a proliferative advantage of one or more of the myeloid differentiating cell lines. For practical purposes and therapeutic considerations, chronic myelomonocytic leukemia is viewed now as a composite disorder along with atypical chronic myeloid leukemia and juvenile myelomonocytic leukemia. PMID- 14560777 TI - Pathology of the myeloproliferative diseases. AB - The classification of myeloid neoplasms now includes CMPD, mixed CMPD/ MDS, MDS, and acute myeloid leukemias. CMPD and CMPD/MDS, both clonal stem cell diseases, share myeloproliferative features, including typical hypercellular marrows, organomegaly, and cell lineage maturation. The CMPD generally differ by which myeloid cell lineage dominates hematopoiesis, and the main diseases include CML, PV, ET, and CIM. The mixed CMPD/MDS disorders also show dysplastic features and variable amounts of effective hematopoiesis; these disorders include CMML, JMML, and atypical CML. Given the overlap in morphology among these diseases, correlation with clinical, hematologic, and cytogenetic/molecular genetic findings is imperative for precise classification. PMID- 14560778 TI - Cytogenetics of chronic myeloproliferative disorders and related myelodysplastic syndromes. AB - The only MPD associated with any specific chromosome anomaly is CML, which is linked with t(9;22)(q34;q11.2) or a variant of this anomaly. An association exists for del(13)(q12q14) and CIMF; t(5;12)(q33;p13) and CEL; and del(20q11), +8, and +9 and PV, but these anomalies can be seen in various hematologic malignancies. The most common chromosomal anomalies among MPD in order of frequency are t(9;22)(q34;q11.2), -Y, +8, +9, -7, del(20) (q11q13), del(13)(q12q14), del(5)(q13q33), and del(12)(p12). FISH techniques are useful for MPD to study inadequate bone marrow or blood specimens and to monitor disease status among patients with known chromosome anomalies, but they are not more sensitive than conventional chromosome studies. PMID- 14560779 TI - Classification and molecular biology of polycythemias (erythrocytoses) and thrombocytosis. AB - In this article, polycythemic disorders are classified based on the current understanding of biology of erythropoieses and divided into primary and secondary polycythemias. Special emphasis is given to recently uncovered molecular bases of newly described congenital polycythemic disorders. This clarification of the pathophysiology of some of the congenital polycythemic states has obvious utility for more accurate diagnosis and rational prognostic determination. The molecular basis of congenital thrombocytoses is only beginning to be uncovered. In contrast, the molecular bases of polycythemia vera and essential thrombocythemia remain unknown, thus their diagnostic criteria are imprecise and their treatment remains largely empirical. The central premise of this article is that deciphering the molecular basis of human diseases leads to improved understanding of hematopoiesis, precise diagnosis, and the potential for development of a specific therapy. PMID- 14560780 TI - Chronic myeloid leukemia. AB - Chronic myeloid leukemia is a clonal myeloproliferative disorder of a pluripotent stem cell with a specific cytogenetic abnormality, the Philadelphia chromosome, involving myeloid, erythroid, megakaryocytic, B lymphoid, and sometimes T lymphoid cells but not marrow fibroblasts. Advances in cell biology and molecular genetics and a plethora of biochemical, cytogenetic, and molecular data of clinical relevance have yielded much new information regarding this disease. This article reviews the hematologic and clinical aspects of chronic myeloid leukemia; discusses the pertinent aspects of the advances in understanding of the cytogenetics and molecular biology of the disease; and reviews treatment programs employing busulfan, hydroxyurea, interferon, and marrow transplantation, which still are clinically important and relevant despite the development of the exciting new drug imatinib mesylate, a new paradigm for cancer chemotherapy in general. PMID- 14560781 TI - Essential thrombocythemia. AB - Essential thrombocythemia is a distinct clinical entity within the spectrum of myeloproliferative disorders. There is as yet no pathognomonic diagnostic test, and patients who currently fall into the category of essential thrombocythemia are likely to be heterogeneous. This article discusses diagnostic criteria, clinical features, prognosis, and management. PMID- 14560782 TI - Polycythemia vera. AB - The differential diagnosis of an elevated hematocrit and the criteria for the diagnosis of polycythemia vera present little or no problem; however, there is not a consensus on therapy. Spivak likened this to a conundrum--"an intricate and difficult problem." Nonetheless, it can be argued that on the basis of the following criteria--life expectancy, the absence of toxicity, and long remissions an average of 3.1 years or a median of 2 years--and with acute leukemia no more common than in other regimens except phlebotomy alone (a regimen that cannot be sustained), 32P should be the treatment of choice except in pregnant women. Others, but not all, share this view. This is in contrast to the statement, "Thus chemotherapy treatment of [polycythemia vera] patients is not as easy, innocuous, and well tolerated as it is generally believed". Patients treated with phlebotomy alone were subjected to an unacceptably high incidence of early thrombotic events. Unavailability of pipobroman eliminates this choice. PMID- 14560783 TI - Myelofibrosis with myeloid metaplasia. AB - MMM is a chronic myeloproliferative disorder characterized by bone marrow fibrosis and neoangiogenesis, constitutive release ofa high number of CD34+ stem cells from the bone marrow, and extramedullary hematopoiesis. It presents with heterogeneous clinical features in which anemia and progression to symptomatic splenomegaly dominate. The pathogenesis is undefined, but the dual action of deregulation of the bFGF pathway may influence myeloproliferation, myelofibrosis, and neoangiogenesis. Animal models suggest that chronic exposure to high doses of thrombopoietin or impairment of the capacity of megakaryocytes to differentiate into platelets, as occurs in the GATA-1(low) mice, is a necessary event for myelofibrosis. Allogeneic stem cell transplantation offers a chance of cure, and low conditioning regimens may extend the age of transplantable patients with lower mortality. Autologus stem cell transplantation and splenectomy are risky procedures that may be considered in patients with advanced disease when conventional therapies for correcting anemia (danazol, recombinant human erythropoietin, or cyclosporine) or chemotherapy for splenomegaly and myeloproliferation (hydroxyurea or interferon alfa) have failed. Thalidomide has been tested in numerous series, and its capacity to improve anemia and thrombocytopenia while reducing splenomegaly has been documented. PMID- 14560785 TI - Allogeneic hematopoietic stem cell transplantation for myeloproliferative disorders and myelodysplastic syndromes. AB - Allogeneic SCT is the most effective method to achieve cure in patients with MPD and MDS. This approach is associated with significant risk of morbidity (eg, GVHD) and TRM, although the incidence and severity vary based on donor and recipient characteristics. For young patients with HLA-matched donors, SCT is the preferred therapy. Efforts to improve outcome for older patients and for patients with alternative donors have led to decreased treatment-associated complications with associated better long-term DFS. PMID- 14560784 TI - Mast cell proliferative disorders: current view on variants recognized by the World Health Organization. AB - The term mastocytosis covers a heterogeneous group of disorders characterized by the abnormal growth and accumulation of MCs in one or more organ systems. Clinical symptoms occur from the release of chemical mediators or pathologic infiltration of MCs. CM typically presents as UP and is a benign disease confined to the skin. In many cases, particularly in children, this disease regresses spontaneously. By contrast, SM is a clonal persistent disease of MC-committed or precommitted hematopoietic progenitors. In most of these patients, the transforming c-kit mutation Asp-816-Val is detectable. The clinical course in SM is variable. Many cases remain in an indolent stage over decades. In a few patients, significant organopathy is found, and this may lead to the diagnosis of aggressive SM. In other patients, AHNMD is diagnosed. MCL is a rare form of SM characterized by leukemic spread of MCs in the marrow and blood and rapid progression. In contrast to indolent SM, patients with MCL or aggressive SM often present without UP-like skin lesions. Patients with indolent SM should be treated with mediator-targeting drugs but not with cytoreductive drugs. By contrast, patients with aggressive SM or MCL are candidates for cytoreductive therapy. Some patients with aggressive SM may benefit from IFNalpha2b. Patients with rapid progression or MCL are candidates for more aggressive treatment, including cladribine, chemotherapy, and stem cell transplantation. In patients with SM AHNMD, the SM should be treated as if no AHNMD is present, and the AHNMD should be treated as if no SM had been diagnosed. PMID- 14560786 TI - Myeloproliferative and myelodysplastic syndromes: the future. AB - The incidence of chronic MPD and MDS could be reduced considerably if relevant environmental factors could be identified and eliminated, but this seems an unlikely prospect for the foreseeable future. More probable is the likelihood that the molecular basis of the various chronic MPD gradually will be elucidated such that specific inhibitory molecules analogous to imatinib may be designed for each disease or each subtype of disease. Combinations of inhibitory molecules may prove especially useful. There is ample scope for improving the clinical results of allogeneic stem cell transplantation, which in theory could "cure" most patients with MPD or MDS. In this regard, reduced-intensity conditioning allogeneic stem cell transplants seem promising. One possibility is identification and exploitation of the basic mechanism underlying the graft versus-leukemia effect for eradication of minimal residual disease without the need for allografting. PMID- 14560787 TI - Evaluation of the mastascanelite image analysis system for measuring zones of inhibition in disc diffusion susceptibility tests. AB - In this evaluation a mastascanelite image analysis system is compared with manual measurement of disc diffusion inhibition zones. Data for 213 diverse organisms and a total of 1679 organism/antibiotic combinations gave an overall correlation coefficient of 0.988. The mean difference between readings was +0.425 mm, with 95% confidence limits of +/-2.94 mm, and the majority of scanned zones (97.51%) fell within +/-3 mm of the manual measurement. The mastascanelite system forms part of a laboratory suite and was found to be objective, accurate and rapid, reading and interpreting each plate in less than a second. Interfacing to the laboratory computer system facilitated data handling and performance control. PMID- 14560788 TI - Streptococcus grouping latex kits: evaluation of five commercially available examples. AB - This study compares a recently introduced latex agglutination test for the serogrouping of beta-haemolytic streptococci against four internationally used commercial kits. The new kit is Prolex-Blue (Pro-Lab Diagnostics) and the comparators are Streptex (Murex), PathoDx (DPC), Streptococcus Grouping kit (Oxoid) and Prolex-White (Pro-Lab Diagnostics). A total of 302 consecutive clinical isolates are tested against all five kits, following the individual manufacturer's protocol, for both accuracy and speed. In addition, the data produced permits determination of the strengths or weaknesses of the kits against individual serotypes. Prolex-Blue proved to be both accurate and rapid, with a sensitivity of 99% and a specificity of 100%. Furthermore, average time to agglutination was substantially less than achieved by three of the other four kits evaluated. PMID- 14560789 TI - Lipaemia: an overrated interference? AB - Reagent method sheets for analysis of common serum analytes often highlight the possibility of interference from lipaemia but the information given is often brief and may not be instrument-specific. Thus study assesses the degree of interference from lipaemia in a range of common serum analytes on the Bayer Opera (with a serum blank) using a commercial polymer, LipoClear, as a lipid-clearing agent. Serum samples (mean serum triglyceride 6.89 [range 0.58-28.4] mmol/L) are analysed for 14 common chemistry analytes and the results compared before and after treatment with LipoClear. Results showed no significant critical differences in analyte values before and after treatment, except for an expected fall in total protein, phosphate, cholesterol and triglyceride concentrations. Most of the common analytes in use on the Bayer Opera are not subject to interference from lipaemia; however, we recommend that where method sheets indicate interference from lipaemia then this should be quantified for the analyte in question. PMID- 14560790 TI - Impaired nitric oxide production, brachial artery reactivity and fish oil in offspring of ischaemic heart disease patients. AB - The offspring of coronary heart disease (CHD) patients are at particularly high risk for developing CHD. Endothelial dysfunction is present in the majority of CHD and atherosclerosis patients. Fish oil, rich in n-3 fatty acids has been shown to augment endothelium-dependent vasodilatation in human peripheral and coronary arteries. The aims of this study are to investigate presence of endothelial dysfunction determined by the brachial flow-mediated diameter, nitric oxide, plasma lipids and fibrinogen, and the effect of high doses of fish oil on these parameters. Twenty-four healthy offspring of CHD patients (study group) were supplemented with 9 g/day Alsepa fish oil (each gram containing 180 mg EPA and 120 mg DHA), for a period of two weeks. Plasma nitric oxide, urine nitric oxide, fibrinogens and flow-mediated vasodilatation (FMD) were determined prior to fish oil therapy, two weeks into therapy and four weeks after the end of therapy with fish oil. Twelve healthy subjects (control group) with no family history of heart disease were studied as controls (day one only). The offspring had a lower increase in FMD and lower nitric oxide production, compared with the control group. No other parameters varied between the two groups. The administration of fish oil did not result in any changes in the studied parameters. In healthy offspring of CHD patients, early endothelial dysfunction was documented before evidence of atherosclerosis. Ingestion of fish oil over a 13-day period did not improve endothelial dysfunction. PMID- 14560791 TI - CD54 and CD62L expression by lymphoid cells in acute lymphoblastic leukaemia in children. AB - Altered expression or function of adhesion molecules on leukaemic blasts may contribute to the evolution and biological behaviour of acute leukaemia. This work studies the expression of CD54 and CD62L by lymphoid cells and the serum level of the shed form of L-selectin (sL-selectin) in children with acute lymphoblastic leukaemia (ALL) at initial diagnosis and after first remission, and their relationship to disease activity and subtype. The study is conducted on 20 children (age range 2-10 years) newly diagnosed with ALL and admitted to Alexandria University Children's Hospital. Ten apparently healthy children of matched age and sex serve as a control group. Expression of CD54 and CD62L on mononuclear cells is detected by monoclonal antibodies using flow cytometry. Serum sL-selectin is measured by enzyme-linked immunosorbent assay (ELISA). B cell ALL was the most common subtype (45%), followed by T-ALL (35%) and C-ALL (20%). CD54 and CD62L mean cellular expression, as well as serum sL-selectin level, were significantly higher at diagnosis than both after remission and in the control group. Univariate analysis showed that the presence of mediastinal mass, high leucocyte count, central nervous system involvement and low CD54 were significant predictors of mortality in children with ALL. PMID- 14560792 TI - Oxidised and native low-density lipoproteins induce the release of von Willebrand factor from human endothelial cells in vitro. AB - Increased low-density lipoprotein (LDL)-cholesterol is a risk factor for atherosclerosis--a disease in which damage to the endothelium is believed to be an important early step. Increased levels of the endothelial marker von Willebrand factor (vWF) in the plasma of patients with hypercholesterolaemia and atherosclerosis probably reflect this process. In this study we seek to link the established observation that oxidised LDL-cholesterol is cytotoxic to human umbilical vein endothelial cells (HUVECs) in vitro with the common finding of raised plasma vWF in patients with atherosclerosis by incubating HUVECs with physiological/pathological levels of native and oxidised LDL-cholesterol for up to 48 h. Microphotography revealed morphological changes in the HUVECs within 24 h, becoming severe at 48 h, which was mirrored by increased levels of vWF (ELISA) and the release of preloaded radioactive (111)indium tracer into culture supernatants. Our data support and extend the hypothesis that oxidised LDL is directly cytotoxic to HUVECs, and, in addition, provide an important link between in vitro studies and clinical studies where endothelial cell markers such as vWF are increased in the plasma of patients with hypercholesterolaemia and atherosclerosis. PMID- 14560793 TI - Survey of microbial air sampling in the NHS. PMID- 14560794 TI - Infective discitis due to Staphylococcus lugdunensis--a case of missed opportunity. PMID- 14560795 TI - Effect of sample tube type and time to separation on in vitro levels of C reactive protein. PMID- 14560796 TI - Enzyme-linked immunosorbent assay for beta2-glycoprotein I quantitation: the importance of variability in the plastic support. PMID- 14560797 TI - TypA is a virulence regulator and is present in many pathogenic bacteria. PMID- 14560798 TI - Molecular typing of Nigerian Helicobacter pylori isolates by glmM restriction fragment length polymorphism. PMID- 14560799 TI - Approximal caries increment: a three-year longitudinal radiographic study. AB - AIM: To determine, for different age groups, the approximal caries increment in a three-year longitudinal retrospective study and to investigate the relationship between the caries increment and self-reported measures for oral health behaviour, dental knowledge and attitude (cynicism, health concern and motivation). SUBJECTS: 14, 17, 20 and 23-year-olds, who participated in a clinical epidemiological survey. Bitewings were taken of approximately 52% of the participants. Longitudinally, three cohorts were followed radiographically for three years. The self-reported measures were derived from questionnaires with sufficient reliability. RESULTS: For all participants, on average, 3.7 enamel lesions and 1.2 dentine lesions were found. The number of sound surfaces, adequate restorations and the D1FS score showed significant differences over a three-year period, for all age groups. Stepwise regression analysis showed that for the explanation of the change in number of sound surfaces, number of enamel lesions and D1S between the age of 17 and 20, the variable 'health concern' contributed significantly to the regression. The change in D3FS score was explained by the variable 'behaviour', while the change in D1Fs score was explained by both 'behaviour' and 'health concern'. Neither of the dental variables, 'dental knowledge' or 'motivation' contributed significantly. For the ages 14-17 and 20-23 no significant contributions were found. CONCLUSIONS: A considerable caries increment was found in this three-year retrospective radiographic study, which was not related to dental knowledge and motivation, and only for one age group (17-20) to oral health behaviour and health concern. PMID- 14560800 TI - Non-carious cervical lesions in a Nigerian population: abrasion or abfraction? AB - OBJECTIVES: To determine the proportion of non-carious cervical lesions (NCCL) that were either abrasion or abfraction like and to test the validity of the assumption that right-handed patients cause more abrasion on the cervical surfaces of teeth on the left side of the mouth. SETTING: Ile-lfe, Southwestern Nigeria. PARTICIPANTS: 106 first time patients diagnosed as having any form of NCCL. DESIGN: Patients were subjected to intra-oral examination followed by the administration of a questionnaire. The cervical lesions were examined and scored using the tooth wear index designed by Smith & Knight 1984. NCCL was diagnosed as abrasion or abfraction like based on established clinical features plus positive history of possible associated aetiological factors. OUTCOME MEASURES: Abrasion and abfraction like lesions. RESULTS: Abrasion was the diagnosis in 630 (62.3%) of the 1,012 tooth sites with NCCL that had no associated occlusal surface wear facet. This constituted about two-thirds of the NCCL. Three hundred and eighty two (37.7%) of the NCCL occurred in association with an occlusal surface wear facet and presented the typical wedge shaped defects with sharp margins characteristic of abfraction. These abfraction like lesions constituted about one third of the NCCL. 50.8% of abrasive NCCL in right-handed brushers were located on the left side of the mouth. CONCLUSIONS: About one-third of NCCL in the present study presents an abfraction component, and two-thirds abrasion. Although, right-handed brushers had more severe lesions on the opposite side of the mouth, the difference was not statistically significant. PMID- 14560802 TI - Global goals for oral health 2020. PMID- 14560801 TI - Children's dental health: British expatriate residents in Tokyo, Japan. AB - AIM: To identify in expatriate British parents resident in Tokyo, Japan, their levels of dental health knowledge, patterns of dental health behaviour, and caries status of their children compared to Japanese children. METHODS: 223 parents with children at one school were asked to complete a structured questionnaire. Data were collected on: parental knowledge and behaviours with respect to caries prevention, and sweet consumption of the children. The caries status of the children was established by a clinical examination, using standard criteria. RESULTS: The questionnaire response rate was 66.8%. The mean age of the children was 7.6 years (range 3-11 years). The responses related to 51 British, 21 Japanese, and 20 children who had British and Japanese mixed parents. 33% of British and none of Japanese children had sweet foods frequently. Fluoridated toothpaste was used by 67% of British and 10% of Japanese children. British parents knew about dental caries more than Japanese. 13% of British and 38% of Japanese children were found in the higher caries experience group. CONCLUSION: Parental access to information in the UK and to fluoridated toothpaste in Japan could have helped to prevent dental caries in these expatriate British children. PMID- 14560803 TI - Oral health knowledge, attitudes and behaviour of children and adolescents in China. AB - OBJECTIVES: A national representative study to describe oral health behaviour, illness behaviour, oral health knowledge and attitudes among 12-year-old and 18 year-old Chinese, to analyse the oral health behaviour profile of the two age groups in relation to province and urbanisation, and to assess the relative effect of socio-behavioural risk factors on dental caries experience. METHODS: The total number of 4,400 of each age group were selected and data were collected by clinical examinations (WHO criteria) and self-administered structured questionnaires. RESULTS: 44.4% of the respondents brushed their teeth at least twice a day but only 17% used fluoridated toothpaste. Subjects who saw a dentist during the previous 12 months or two years were 31.3% and 35.3% for 12-year-olds and 22.5% and 20.2% for 18-year-olds, respectively. Nearly one third (29%) of 12 year-olds and 40.5% of 18-year-olds would visit a dentist in case of signs of caries but only when in pain. Nearly half of the participants (47.2%) had never received any oral health care instruction. Significant variations in oral health practices were found according to province and regular dental care habits were more frequent in urban than in rural areas. The risk of dental caries was high in the case of frequent consumption of sweets and dental caries risk was low for participants with use of fluoridated toothpaste. CONCLUSION: Systematic community oriented oral health promotion programmes are needed to target lifestyles and the needs of children, particularly for those living in rural areas. A prevention oriented oral health care policy would seem more advantageous than the present curative approach. PMID- 14560804 TI - International profiles of dental hygiene 1987 to 2001: a 19-nation comparative study. AB - AIM: The purpose of this international longitudinal study is to examine patterns and monitor trends and changes in dental hygiene. METHOD: Information was collected from national dental hygienists' associations through surveys conducted in 1987, 1992, 1998 and 2001. Sample size increased from 13 countries in 1987 to 22 by 2001--of which 19 were included in the analysis. RESULTS: Overall, characteristics of the profession were remarkably similar; most noteworthy was the scope of dental hygiene clinical practice. Regarding historical development, educational programmes and professional organisation, the profession was more similar than dissimilar. Greater variation was evident regarding numbers, distribution, regulation, workforce behaviour, predominant work setting, and remuneration. Over the relatively short 14-year period, several observations were of particular interest: marked increase in the supply of dental hygienists, accompanied by a decline in their ratio to populations and to dentists and a high workforce participation rate; increase in baccalaureate dental hygiene programmes, with a gradual shift from the diploma as the entry-level qualification; and increase in scope of practice and professional autonomy, including for Europe and North America in particular, a decline in mandated level of work supervision and a slight but gradual increase in independent practice. CONCLUSION: By 2001, the profiles reflected the vast majority of the world's population of dental hygienists. Rate of change varied across the countries examined; however, the nature of the change overall was consistent, resulting in a continuing homogeneity in the profession worldwide. Observed trends, changes and persistent issues have implications for service accessibility and technical efficiency and should continue to be monitored. PMID- 14560805 TI - Minimally invasive periapical curettage of foreign materials in periapical lesions using a fiberscope. AB - The design and development of a new fine fiberscope (1.0 mm outer diameter) is described, that includes an irrigation system for endoscopic use. This effectively allows visualisation of fields involved in periodontal disease. This fiberscope has been used in our clinic for seven years to diagnose and treat periapical and periodontal lesions through fistulae and periodontal pockets. The fiberscope has been applied to endodontic surgery for the inspection of root apices and the denuded root surfaces, in addition to the cut root face, rootend cavity and root-end filling after root-end resection. The advantages of endoscopy compared to surgical microscopy are portability and easy adjustment of direct viewing angle to examine around corners without the use of additional mirrors. Furthermore, the fiberscope can be utilised through the sinus tract or a minimal incision without the need for surgical flaps to observe the root surface, periapical tissue and foreign materials before, during and after curettage. The system requires little local anaesthesia, no large incisions, no opening flaps and no sutures. As a result, postoperative symptoms are minimised. Periapical curettage using a fiberscope represents a useful and minimally invasive surgical procedure. PMID- 14560806 TI - Heat shock and UV-B-induced DNA damage and mutagenesis in skin. AB - There is evidence that heat pre-treatment protects cultured human keratinocytes and normal murine and human skin from ultraviolet (UV) radiation-induced cell death. It has been suggested that heat-shock proteins (hsps), particularly hsp72, are involved in this effect. Hsps are expressed in response to various types of stress, such as UV radiation. Whether heat shock interferes with the repair of UV induced DNA damage and whether this can be regarded as a protective mechanism is poorly understood and needs further experimental investigation. This review gives an overview of the current state of research in the area. PMID- 14560807 TI - On the Paterno-Buchi reaction of chiral phenylglyoxylate esters with furan derivatives. AB - The reaction of (S)-1-methylpropylbenzoylformate with furan gave (1'S)-1' methylpropyl (1S,5R,6R)-6alpha-phenyl-2,7-dioxabicyclo[3.2.0]hept-3-en-6beta carboxylate with de = 15%. The reaction of (S)-2-methylbutyl benzoylformate with furan gave (2'S)-2-methylbutyl (1SR,5RS,6RS)-6alpha-phenyl-2,7 dioxabicyclo[3.2.0]hept-3-en-6beta-carboxylate as a mixture of stereoisomers. (1R,2S,5R)-5-Methyl-2-(1-methylphenylethyl)cyclohexyl benzoylformate gave (1R,2S,5R)-5-methyl-2-(1-methylphenylethyl)cyclohexyl (1R,5S,6S)-6alpha-phenyl 2,7-dioxabicyclo[3.2.0]hept-3-en-6beta-carboxylate with de = 95%. The stereochemical behaviour can be explained considering the energy gap between the biradical intermediates in the coupling reaction. When the reaction was performed in the presence of zeolite, the diastereoisomeric excess increased (37, 18 and 98%, respectively). The reaction of (S)-1-methylpropylbenzoylformate with 2 methylfuran gave (1'S)-1'-methylpropyl (1S,5R,6R)-1-methyl-6alpha-phenyl-2,7 dioxabicyclo[3.2.0]hept-3-en-6beta-carboxylate (yield: 12%) with de = 94%. The main product was (1'S)-1'-methylpropyl (1SR,5RS,6RS)-3-methyl-6alpha-phenyl-2,7 dioxabicyclo[3.2.0]hept-3-en-6beta-carboxylate as a mixture of diastereoisomers. The reaction of (S)-2-methylbutyl benzoylformate with 2-methylfuran gave (2'S)-2 methylbutyl (1S,5R,6R)-1-methyl-6alpha-phenyl-2,7-dioxabicyclo[3.2.0]hept-3-en 6beta-carboxylate (yield: 9%) with de = 10%. The main product was (2'S)-2 methylbutyl (1SR,5RS,6RS)-3-methyl-6alpha-phenyl-2,7-dioxabicyclo[3.2.0]hept-3-en 6beta-carboxylate as a mixture of stereoisomers. The stereochemical behaviour can be explained on the basis of the energy gap between the possible biradical intermediates. The reaction of (S)-1-methylpropylbenzoylformate and (S)-2 methylbutyl benzoylformate with 2-furylmethanol gave the corresponding adducts as a mixture of diastereoisomers. The reaction of (S)-1-methylpropylbenzoylformate and (S)-2-methylbutyl benzoylformate with 2-furylphenylmethanol gave the corresponding adducts with high diastereoselectivity. In this case, the stereochemical behaviour is determined by the presence of the hydroxy group in a molecule with a preferntial conformation. PMID- 14560808 TI - Studies on the photochemistry of 1,7-diphenyl-1,6-heptadiene-3,5-dione, a non phenolic curcuminoid model. AB - The comparative photostability of curcumin 1, and two non-phenolic curcuminoids: 1,7-diphenyl-1,6-heptadiene-3,5-dione 2 (unsubstituted curcumin) and dimethylcurcumin 3 in non-degassed dilute solutions (approximately 3-5 x 10(-5) mol l(-1)) has been established by UV-visible absorption spectroscopy; disappearance quantum yields were measured. The similar behavior of the three studied curcuminoids is indicative of only a moderate role of phenol groups in the photodegradation process. Structural analysis of the photodegradation products of compound 2 in more concentrated solution (approximately 3.6 x 10(-3) mol l(-1)) shows formation of benzaldehyde, cinnamaldehyde, 2'-hydroxy-5',6' benzochalcone 4, flavanone 5 and some other unidentified photoproducts. Flavanone 5 is formed by irradiation of chalcone 4. It represents a unique example of photochemical conversion of a diarylheptanoid molecule into a flavonoid, another very important class of natural products. PMID- 14560809 TI - Preparation of silver nanoprisms using poly(N-vinyl-2-pyrrolidone) as a colloid stabilizing agent and the effect of silver nanoparticles on the photophysical properties of cationic dyes. AB - The synthesis of silver nanoprisms in aqueous solution using poly(N-vinyl-2 pyrrolidone) (PVP) with different molecular weights (29, 55 and 1300 kg mol(-1)) as a stabilizing agent is described. Low molecular weight PVP (55 kg mol(-1)) is indicated for the preparation of a stable blue solution containing Ag nanoprisms via extended irradiation of a yellow colloidal solution of nanospheres with polychromatic visible light. The fluorescence properties of some cationic dyes (acridine, 9-aminoacridine, Nile Blue and auramine), free and bound to poly(methacrylic acid), have been studied in the presence of different shaped Ag colloidal particles in solution. Only auramine displays an amplification of the fluorescence signal with increasing colloid concentration, while the fluorescence of the other dyes is quenched by interaction with the polymer-stabilized metal nanoparticles. PMID- 14560810 TI - Photodynamic activity of monocationic and non-charged methoxyphenylporphyrin derivatives in homogeneous and biological media. AB - A novel 5-[4-(trimethylammonium)phenyl]-10,15,20-tris(2,4,6 trimethoxyphenyl)porphyrin iodide (2) has been synthesized. A positive charge was incorporated at a peripheral position to increase the amphiphilic character of the structure. The photodynamic effect of the cationic porphyrin 2 was compared with that of non-charged 5-(4-aminophenyl)-10,15,20-tris(2,4,6 trimethoxyphenyl)porphyrin (1), both in a homogeneous medium bearing photooxidizable substrates and in vitro on the Hep-2 human larynx carcinoma cell line. Absorption and fluorescence spectroscopic studies in different media show that 2 is essentially unaggregated in solution, and also in human cells. The singlet molecular oxygen, O2(1delta(g)), production was evaluated using 9,10 dimethylanthracene in N,N-dimethylformamide, yielding phi(delta) values of approximately 0.66 for both porphyrins. The addition of beta-carotene suppresses the O2(1delta(g))-mediated photooxidation. L-Tryptophan and guanosine 5' monophosphate were used as biological substrate models. Porphyrin 2 sensitizes the decomposition of both compounds faster than does 1. In the biological medium, no dark cytotoxicity was observed, even though a high porphyrin concentration (10 microM) and a long incubation time (24 h) were employed. Cell treatments were performed with 5 microM of porphyrin for 24 h. Under these conditions, the uptake of porphyrin 2 into Hep-2 was about 3 times higher than that of 1. Cell survival after irradiation with visible light was dependent upon both the light exposure level and intracellular sensitizer concentration. Thus, a higher photocytotoxic effect was found for porphyrin 2 in comparison to 1. These results show that the amphiphilic monocationic porphyrin 2 could be a promising model for phototherapeutic agents with potential applications in tumor cell inactivation by photodynamic therapy. PMID- 14560811 TI - Photophysical behaviour of an opp-dibenzoporphyrin (2,12-diethyl-3,13 dimethyldibenzo[g,q]porphyrin) in micelles and organic solvents. AB - Photophysical parameters relevant to photodynamic therapy have been studied for a novel lipophilic opp-dibenzoporphyrin (DBP), 2,12-diethyl-3,13 dimethyldibenzo[g,q]porphyrin, in N,N-dimethylformamide and TX-100 micelles. The structure of DBP is intermediate between a porphyrin and a phthalocyanine and is associated with higher molar extinction coefficients in the red Q-bands than those found in the haematoporphyrin derivative currently used for photodynamic therapy. The ultrafast measurements on DBP in DMF revealed a fast (ps) lifetime for the second excited singlet state. Observed lifetimes of the first excited singlet state were found to be similar in DMF and TX-100 (12.2 and 14.7 ns), but excited triplet lifetimes were different in the two solvents (0.46 and 2.86 micros). The fluorescent quantum yields of DBP in DMF and TX-100 were twice that of free-base tetraphenylporphyrin and the singlet oxygen quantum yield in DMF and TX-100 was high (0.56-0.65). The combination of stable chemical structure, stronger red-absorption, high singlet oxygen quantum yields, and high fluorescent quantum yields suggests that DBP is an potential chromophore for applications in photodynamic therapy. PMID- 14560812 TI - Investigation of singlet oxygen reactivity towards propofol. AB - The reaction between the anaesthetic agent 2,6-diisopropylphenol (propofol, PPF) and singlet oxygen (1O2) has been investigated in aqueous solution by means of HPLC, GC, absorption spectroscopy and laser flash photolysis with infrared luminescence detection. The rate constants for the physical and chemical quenching of 1O2 by PPF (kPPF) are found to be 2.66 x 10(5) M(-1) s(-1) and approximately 3.2 x 10(6) M(-1) s(-1) in CD3OD and D2O-CD3OD (75:25 v/v), respectively. The reaction of propofol with singlet oxygen produced by light irradiation of Rose Bengal leads essentially to two reaction products, 2,6 diisopropyl-p-benzoquinone and 3,5,3',5'-tetraisopropyl-(4,4')-diphenoquinone that are unambiguously identified from comparison with authentic samples. PMID- 14560813 TI - Transformation of 4-tert-octylphenol by UV irradiation and by an H2O2/UV process in aqueous solution. AB - The transformation of the organic pollutant 4-(1,1,3,3-tetramethylbutyl)phenol (4 tert-octylphenol; OP) upon irradiation at 253.7 nm and by hydroxyl radicals generated by the photolysis (lambda(exc) = 253.7 nm) of hydrogen peroxide in aqueous solution has been studied. The quantum yield of direct OP photolysis in pure aqueous solution was evaluated to be 0.058 +/- 0.004 in aerated conditions ([O2] = 272 microM). The rate of photoreaction depends on oxygen concentration; it increases with increasing [O2]. 4-tert-Octylcatechol has been identified as one of the degradation products, together with a dimeric structure. The probable mechanism of OP photolysis involves photoejection of an electron from the singlet state, leading to the formation of the 4-tert-octylphenoxyl radical. In the presence of hydrogen peroxide, the degradation of octylphenol by hydroxyl radicals has been observed. The second-order rate constant was found to be (6.4 +/- 0.5) x 10(9) M(-1) s(-1) by direct measurement at various high concentrations of hydrogen peroxide and competitive kinetic measurements using atrazine as the competitor. The degradation products are 4-tert-octylcatechol, again, and 2 hydroxy-5-tert-octylbenzoquinone. The later product may arise from the oxidation of 4-tert-octylcatechol by hydrogen peroxide or from a subsequent reaction of hydroxyl radicals with 4-tert-octylcatechol. Kinetic modelling when using either purified water or natural water successfully simulated the elimination of 4-tert octylphenol by UV and H2O2/UV processes. PMID- 14560814 TI - The complex between 9-(n-decanyl)acridone and bovine serum albumin. Part 2. What do fluorescence probes probe? AB - Factor analysis indicates that the fluorescence spectrum of 9-(n-decanyl)acridone (NDA), when bound to Bovine Serum Albumin (BSA), can be described quite adequately as the sum of two spectra, attributed to a "free" and a "bound" species. Kinetic evidence indicates that upon electronic excitation the system undergoes a net increase in free NDA, relative to the equilibrium distribution in the ground state, which would be consistent with Lewis acid sites on BSA being responsible for the binding. The system does not attain a position of equilibrium during the duration of the excited singlet state. This permits the determination of excited state rate constants for binding and unbinding of NDA on BSA, as well as the decay constants for the two forms of the probe. PMID- 14560815 TI - Degradation of surfactants by hydroxyl radicals photogenerated from hydroxoiron(III) complexes. AB - The Fe(III)-photoinduced oxidation of anionic lauryl sulfate (LS-) and cationic cetyltrimethylammonium (CTA+) surfactants has been investigated in aqueous solution. Competition experiments using 2-propanol showed that the initial rate of disappearance is proportional to the concentration of the photogenerated HO* radicals scavenged by the surfactants (the degradation of lauryl sulfate involves attack by HO* only) and no direct photoinduced charge-transfer reaction occurs between the Fe(III) species and the surfactant ions. Ageing of the Fe(III) solution did not significantly influence the efficiency of photodegradation in air-saturated systems. Conversion of Fe(III) to Fe(II) in aerated solution reached a steady-state level of ca. 50% after 2 h irradiation. In nitrogen saturated systems, the rate of surfactant oxidation decreased due to the total reduction of Fe(III). Addition of H2O2 doubled the quantum yield of the disappearance of both detergents as a result of the photo-Fenton reaction. The photoinduced oxidation of both surfactants was most efficient in acidic solutions of pH 2-3, without H2O2, and for the photo-Fenton system; the quantum yields are phi(NaLS) = 0.011, phi(CTAB) = 0.012 without H2O2, and phi(NaLS) = 0.024, phi(CTAB) = 0.027 in the photo-Fenton system with irradiation at 366 nm. For the disappearance of 4 x 10(-4) M detergent, due to the first oxidation step, 4 h of irradiation (at pH 2.6) is sufficient, whereas 100% mineralization of the total organic carbon content requires prolonged photolysis for at least 10 h. The formation of carbon dioxide dramatically accelerated after a 2 h induction period (1 h in the photo-Fenton system), indicating the cleavage of the long hydrocarbon chains to shorter intermediates in the first stage of the mechanism. The following step is total mineralization of these smaller compounds, which were identified as mostly hydroxy acids via GC-MS. PMID- 14560816 TI - Evidence for diffusion-controlled electron transfer in exciplex formation reactions. Medium reorganisation stimulated by strong electronic coupling. AB - Diffusion-controlled rates of formation were found from the temperature dependence of apparent quenching rate constants for exciplexes, when the driving force of excited-state electron transfer -0.1 < deltaG(ET)* < +0.1 eV. This is inconsistent with the conventional mechanism of electron-transfer reactions, involving preliminary reorganisation of the medium and reactants, and provides strong support for the mechanism of medium reorganisation stimulated by strong electronic coupling of locally excited and charge-transfer states. PMID- 14560817 TI - Focal seizures and EEG: prognostic considerations. AB - The goal of this study was to determine whether the EEG could predict if patients with focal seizures would eventually be uncontrolled (U), more than two seizures per month, or be controlled (C), fewer than two seizures per year. Using these latter criteria, U and C patients were randomly selected from our files, 150 in each of these two groups; 804 EEGs were found in the U and 674 in the C group. Excluded were patients with generalized epilepsy and also the benign epilepsies of childhood. Age was an important factor since patients 5-18 yrs old were more often under good control (C group), and those 21-48 yrs of age were more often under poor control (U group). With rare exceptions, focal spikes and focal slow waves in every area were much more often seen in U patients than C patients. No spikes or rare spikes appeared especially in the controlled patients, and many or very many spikes in the uncontrolled patients, mainly on the first or second EEG. No slow waves or a mild degree of slowing was seen especially in the C patients, while greater degrees of slow wave abnormalities were noted much more often in the U patients. For conditions to identify the C group, the best predictors were no spikes or rare spikes, especially on the first or second EEG, properly identifying two-thirds of the C patients and misidentifying only one-fourth of the U group. To identify the U patients on any EEG, many or very many spikes at any location or frontal spikes correctly designated a U patient in 84%, but incorrectly predicted a C patient as uncontrolled in 29%. For the first or second EEG, these values were 61% and 21%. Thus, the EEG, especially by its first or second record, can predict well the probable future of the uncontrolled patient, so that extraordinary means may be instituted early to avoid a deteriorating condition. When complete EEGs are done with sleep records and the results are quantified, reasonable prediction of eventual outcome can usually occur that is much more timely than waiting over 9 yrs, using only clinical data. PMID- 14560818 TI - Effects of stimulus modality and response mode on the P300 event-related potential differentiation of young and elderly adults. AB - The P300 event-related brain potential (ERP) was examined in 14 young (20 - 29 years of age) and 16 elderly (60 - 82 years of age) adult subjects during the performance of auditory and visual discrimination tasks requiring silent counting or key pressing in response to target stimuli. P300 latencies were longer in elderly (vs young) adults and in visual (vs auditory) tasks, and visual tasks elicited larger P300 amplitudes than auditory tasks in both age groups. Neither stimulus modality nor response mode affected P300 differentiation of young and elderly subjects. Steeper P300 anterior-posterior scalp amplitude gradients were seen in the young (vs elderly) adults, regardless of stimulus or response type. Examination of inter-subject variability with the coefficient of variation (CV) statistic found the lowest (i.e., best) CV values to be exhibited in the visual task requiring the counting of target stimuli. Implications of the findings are discussed in relation to P300 applications in the clinical assessment of dementia and aging-associated cognitive alterations. PMID- 14560819 TI - Olfactory event-related potentials in normal subjects and patients with smell disorders. AB - We developed a device to record olfactory event-related potentials (OERP) from the human scalp. Methyl-cyclopentenolone was used as the odorant element. A total of 50 stimuli were delivered. Each stimulus lasted 0.5 sec and was delivered once every three inspirations. Normal OERPs were obtained with this device. The positive peak latencies were approximately 350 msec (P1) and 700 msec (P2), respectively. OERPs were also recorded in 40 patients with smell disorders. A positive response at about 300-400 msec was recorded in 7 patients (all females, 15-59 years old). The other 33 patients showed no response. The high potential area of this positive peak was located in the centro-occipital region of the scalp. The latency and the high potential area of this peak were similar to P1 recorded in normal subjects. The source of this peak was considered identical to that of P1. This may be a response to the trigeminal nerve during odor administration. P2 was not recorded in the patients with smell disorders. P2 may therefore be a response to the olfactory nerve. PMID- 14560820 TI - Apneic seizures in infants: role of continuous EEG monitoring. AB - Respiratory compromise is not uncommon in epileptic seizures. However, pure apneic seizures are rare. In this study, we report 10 children who presented with pure apneic seizures. All the children were admitted because of apneic events. Seizures were also considered in the differential diagnosis. Six patients had nonspecific findings consisting of multifocal interictal epileptiform activity with no event correlation. Continuous 24-72 hours electroencephalogram (EEG) was performed in all patients to rule out apneic seizures. Ictal EEG showed high correlation with the apneic episodes, confirming the diagnosis of apneic seizures. Our study suggests that continuous EEG monitoring is essential in the diagnosis and treatment of apneic seizures. PMID- 14560821 TI - Sequential EEG analysis during intermittent photic stimulation in never-medicated patients with schizophrenia. AB - To investigate sequential changes in electroencephalograms (EEGs) during intermittent photic stimulation (IPS), quantitative EEG analysis for the alpha band (related to stimulus frequency) was performed in 18 drug-naive schizophrenic patients and 18 sex- and age-matched control subjects. IPS consisted of white flickers at 10 flashes/sec, lasting 10 sec and repeated six times at 10 sec intervals. The differences between the two groups were assessed for two conditions (i.e., during photic stimulation (PS) and inter-PS). During stimulus, the absolute 9-11 Hz band power of the patient group was higher at the posterior than that at the anterior sites throughout the 10 sec periods. However, this difference between sites was not seen during the first 1 sec in the control group. During the nonstimulus period, posterior dominance of 9-11 Hz band power was prominent in the patient group throughout. In the control group, however, this difference was minor, especially during the latter half of the period. These results suggest that schizophrenic patients show fewer changes in posterior alpha activity during both stimulus and nonstimulus, and this continuity of posterior dominance may reflect hyperarousal, which counteracts any decrease in vigilance throughout the IPS. PMID- 14560822 TI - Time distribution of seizures during long-term video-EEG monitoring. AB - The purpose of this study was to investigate the duration of monitoring, number of seizures recorded, and date and time of occurrence of seizures in different patient groups during video-EEG monitoring (VEEGM). Patients with partial or psychogenic seizures who were admitted to our VEEGM unit between September 1996 and March 2002 were retrospectively evaluated. Duration of monitoring, date and time of occurrence of seizures were identified in each patient. For practical reasons, a day was divided into 3 equal time periods (period 1: 08-16; period II: 16-24 and period III: 24-08 hrs), and the period in which the seizure occurred was noted. Patients were classified in 5 groups: 1-temporal; 2-frontal; 3-parieto occipital; 4-psychogenic and 5-unclassified. Duration of monitoring, the number of seizures recorded and time distribution of seizures were identified in each group and compared statistically. We evaluated 746 seizures in 209 patients. Seizures were more likely to occur in periods I and III in groups 1 (p = 0.004) and 5 (p < 0.001). In group 4 they were more likely to occur in periods I and II (p < 0.001). Duration of monitoring was not statistically different between the groups. However, the number of seizures recorded varied significantly (p = 0.006). Patients in group 5 had the greatest number of seizures and were followed by groups 2, 4, 3 and 1, respectively. Groups also differed according to date of seizure occurrence (p < 0.001). We conclude that the number of seizures recorded and time distribution of seizures during VEEGM are influenced by the lobe of onset of seizures. PMID- 14560823 TI - Orthopedic trauma: is it a subspecialty? PMID- 14560824 TI - Recent advances toward the clinical application of bone morphogenetic proteins in bone and cartilage repair. AB - Inefficient healing of bony and cartilaginous defects is a common situation encountered by orthopedic surgeons. Enhancing the regenerative potential of bone and articular cartilage has the potential for profound applications in treatment of nonunions, large segmental bone and cartilage defects, and arthritis. The bone morphogenetic proteins (BMPs) encode a highly conserved class of signaling factors that possess the ability to induce ectopic cartilage and bone formation in vivo. Bone morphogenetic protein family members are expressed during limb development, endochondral ossification, and early fracture and cartilage repair. Loss-of-function and gain-of-function studies have demonstrated the necessity and sufficiency of these genes, respectively, in regulating both cartilage and bone development. Several recent animal studies have demonstrated the potential of BMPs to enhance spinal fusion, repair critical-size defects, accelerate union, and heal articular cartilage lesions. A limited number of clinical trials using BMPs in human beings have been reported, and these agents are currently available for clinical use within and outside the United States. Current challenges to be met are the development of efficient delivery systems to present BMP proteins or genes to target sites and to enhance their duration and function at these locations. PMID- 14560825 TI - Two or three screws for fixation of femoral neck fractures? AB - This study compares the stability of 3 cannulated cancellous lag screws with that of 2 cannulated cancellous lag screws for fixation of subcapital femoral neck fractures. Using 10 matched pairs of human cadaveric femurs, subcapital femoral neck osteotomies were created, reduced, and then randomized to 1 of the 2 fixation methods. The constructs were tested with anterior loading to 500 N, incremental axial loading from 100 N to 1000 N, and cyclic loading at 1000 N. The specimens stabilized using 3 screws showed greater resistance to anterior loading, less inferior femoral head displacement, and less superior gapping at the osteotomy site. Although 2 screws may be an acceptable fixation method for this fracture type, the addition of a third screw provides supplemental stability and appears justified. PMID- 14560826 TI - Classification of intertrochanteric fractures with computed tomography: a study of intraobserver and interobserver variability and prognostic value. AB - The purpose of this prospective study was to determine the level of interobserver and intraobserver agreement among orthopedic surgeons and radiologists when computed tomography (CT) scans are used with plain radiographs to evaluate intertrochanteric fractures. In addition, the prognostic value of current classifications systems concerning quality of life was evaluated. Sixty-one patients who presented with intertrochanteric fractures received open reduction and internal fixation with compression hip screw. Three orthopedic surgeons and 2 radiologists independently classified the fractures according to 2 systems: Evans Jensen and AO (Arbeitsgemeinschaft fur Osteo-synthesefragen). Fractures were initially graded with plain radiographs and then again in conjunction with CT. Results were analyzed using the (kappa) kappa coefficient. The 36-item Short-Form Health Survey was administered at baseline, 3 months, and 1 year, and results were correlated with fracture grade. Mean kappa coefficients when comparing radiography alone with radiography and CT scan were 0.63 for the AO system and 0.59 for the Evans-Jensen system. Both represent "fair" agreements. Mean overall interobserver kappa coefficients were 0.67 for radiologists and 0.57 for orthopedic surgeons. Radiologists also had higher intraobserver kappa coefficients. No significant relationships were found between follow-up Short Form Health Survey results and intraoperative grading of fractures. When these classification schemes are compared, interobserver agreement does not appear to change dramatically when information from CT scans is added. This may suggest that (1) more data have been provided by CT with greater possibilities for misinterpretation and (2) these classification schemes may not be comprehensive in describing fracture pattern and displacement. Finally, both systems failed to provide any prognostic value. PMID- 14560827 TI - Triplane fracture of the proximal tibia. AB - We report a case of successful treatment of a very rare triplane fracture of the proximal tibia in an adolescent boy. The fracture was internally fixed using cannulated Herbert screws. The final result was excellent. PMID- 14560828 TI - Hip pain in a case of hypophosphatemic osteomalacia. AB - Hypophosphatemic rickets, a rare metabolic bone disease, presents mainly in children but has also been reported in several adults. In this report, we describe the case of a man presenting with hip pain and weakness, both of several months' duration, and tested for hypophosphatemic rickets. The patient was eventually referred to a tertiary-care center, where he was diagnosed with bilateral subtrochanteric femoral stress fractures and severe osteopenia secondary to hypophosphatemic osteomalacia. The patient was treated with closed reduction and internal fixation and vitamin D and phosphorus. Outcomes were good at 7-month follow-up. PMID- 14560829 TI - Injuries associated with fracture of the femur secondary to motor vehicle accidents in children. AB - A retrospective review of the medical records of 149 children < 18 years old who sustained femur fractures secondary to a motor vehicle accident found 159 fractured femurs; 89.3% were closed. One hundred seventy-nine associated injuries occurred in 87 patients (58.4%). Associated fractures were common (50.3%), followed by soft-tissue injuries (18.5%) and head injuries (14%; 3 deaths). The lower limbs were most commonly involved with associated fractures (37.8%), 9 had a "floating knee." The incidence of intra-abdominal injury was 5%, with the liver most commonly lacerated followed by the spleen and intestine. Forty-seven children (43.1%) developed 1 or more complications. The Pediatric Trauma Score ranged from -3 to 11 (73.8% scored > or = 8, while 26.2% scored < 8). Fractures of the femur in children are commonly associated with other injuries, particularly other fractures. PMID- 14560830 TI - Use of the AO femoral distractor in revision total hip arthroplasty. AB - Use of the femoral distractor in trauma settings is well documented in the orthopedic literature. Use of the device extends into adult reconstructive surgery; surgeons use this distractor as an aid in performing difficult revision total hip arthroplasties (THAs). Patients with associated soft-tissue contractures secondary to subacute or chronic dislocation of a THA may present a clinical picture that might make consideration of simple open reduction only a remote possibility. Patients with long-standing Girdlestone resection arthroplasty usually present with soft-tissue deficiencies that render the hip unstable (unless it is at maximal length), or these patients have severe contractures that make reduction difficult after all components are implanted. We present a distractor technique for reduction of prosthetic components in such clinical situations. The femoral distractor is easily applied, and, because of its mechanical effectiveness, it can assist in reducing the "irreducible" hip. PMID- 14560831 TI - Relationships between visual acuity and anomalous head posture in patients with congenital nystagmus. AB - PURPOSE: To determine whether patients with congenital nystagmus and an anomalous head posture have better binocular visual acuity than such patients without an anomalous head posture. PATIENTS AND METHODS: This was an observational case series of prospectively collected data for 125 patients with clinical and oculographically confirmed congenital nystagmus. Clinical data were tabulated using computer software. Statistical analyses compared binocular visual acuity with and without the presence of a clinically evident anomalous head posture and visual acuity with and without associated sensory disease. RESULTS: The mean visual acuity was 20/42 (log of the minimal angle of resolution [MAR], 0.32) in patients with an anomalous head posture and 20/83 (logMAR, 0.62) in patients with no anomalous head posture (P < .001). Among patients with disease of the sensory system, those with an anomalous head posture had a mean visual acuity of 20/55 (logMAR, 0.44) and those without an anomalous head posture had a mean visual acuity of 20/108 (logMAR, 0.73; P < .001). CONCLUSIONS: Visual acuity was found to be significantly better in patients with congenital nystagmus who had an anomalous head posture versus those without such a head posture. Our findings indicate that the presence of an anomalous head posture in a patient with congenital nystagmus correlates with good vision and thus may be considered a positive prognostic sign in a preverbal child. PMID- 14560832 TI - Histopathologic analysis of 232 eyes with retinoblastoma conducted in an Indian tertiary-care ophthalmic center. AB - PURPOSE: To study the histopathologic features of 232 enucleated eyes with retinoblastoma. MATERIALS AND METHODS: Two hundred thirty-two enucleated eyes with retinoblastoma in a tertiary-care institute from 1982 to 2001 were reviewed. Data were collected and analyzed about the type of growth and the presence or absence of vitreous or subretinal seeding, rosettes and fleurettes, necrosis, calcification, iris neovascularization, and invasion of the anterior chamber, iris, choroid, optic nerve, and sclera. Choroidal invasion was graded using a new system. Results were analyzed for statistical significance. RESULTS: The endophytic growth pattern was common in 118 (51%) of the eyes. Vitreous seeds were present in 109 (47%) of the tumors, 23 (10%) of the tumors had subretinal seeds, and 14 (6%) of the tumors had both. Poorly differentiated tumors were present in 134 (58%) of the eyes. Iris neovascularization was noted in 71 (31%) of the eyes and choroidal invasion was observed in 78 (34%) of the eyes. Of these 78 eyes, full-thickness (stage 4) choroidal invasion was present in 51 (65%). Optic nerve invasion was observed in 75 (32%) of the eyes, of which prelaminar involvement occurred in 40 (53%) and postlaminar involvement occurred in 22 (29%). CONCLUSION: A higher incidence of choroidal and optic nerve infiltration was noted among Asian Indian children than among children from the West. This could be due to delayed diagnosis or to a difference in the biological behavior of tumors occurring in the Asian Indian population. PMID- 14560833 TI - Enhanced visualization of capsulorhexis with indocyanine green staining in pediatric white cataracts. AB - PURPOSE: To evaluate the enhancement of the visualization of the anterior lens capsule with indocyanine green (ICG) in pediatric dense, mature cataracts. PATIENTS AND METHODS: Six children (10 eyes) 4 to 9 years old with complete, white cataracts underwent ICG-enhanced cataract surgery. RESULTS: All of the eyes showed a complete continuous curvilinear capsulorhexis and a well-centered intracapsular implant. No surgical or postoperative complications were encountered. CONCLUSION: Staining the anterior capsule with ICG is an excellent way to facilitate performance of an anterior capsulorhexis in pediatric white cataracts. PMID- 14560834 TI - Ocular and systemic findings associated with optic disc colobomas. AB - PURPOSE: To determine the ocular and systemic anomalies associated with optic disc colobomas. PATIENTS AND METHODS: The records of patients with a diagnosis of isolated optic disc coloboma and chorioretinal coloboma with optic disc involvement were retrospectively reviewed. RESULTS: Fifteen patients were included in the study. Of the 30 eyes, the optic disc and choroid were involved in 18, an isolated disc coloboma was present in 5, a normal optic disc was present in 4, and the optic disc could not be identified because of extreme microphthalmia in 1. Of the optic discs outside the fundus colobomas, 1 had an abnormal shape and 1 had optic atrophy and hypoplasia. Eight patients had bilateral but asymmetric involvement of the optic nerve with the coloboma. Fourteen eyes of 9 patients were microphthalmic. Nine eyes of 6 patients had microcornea. One patient had a nonrhegmatogenous retinal detachment at the time of diagnosis. One eye had a retrobulbar cyst with microphthalmia. Eight (53%) of the patients had sensory strabismus. Two patients had unilateral cortical lens opacities. Associated systemic findings were present in 6 (40%) of the 15 patients. CONCLUSIONS: Optic disc colobomas have a wide variety of presentations. Poor visual acuity was observed mostly in eyes with macular involvement and microphthalmia. PMID- 14560835 TI - Services provided for preschool-aged children with suspected amblyopia. AB - BACKGROUND: Little is known about the pattern and variation of care offered to preschool-aged children who have had an abnormal vision screening test. PURPOSE: To evaluate the variations in pediatric eye care services and availability of optometrists and ophthalmologists for preschool-aged patients, referral patterns, and barriers to providing care as perceived by eye care specialists. METHODS: A survey was mailed to 542 ophthalmologists and a random sample of 501 optometrists actively practicing in Michigan. RESULTS: The response rate was 65% (optometrists, 75%; ophthalmologists, 57%). More optometrists than ophthalmologists evaluated preschool-aged children (97% vs 79%; P < .001). Of these, most managed amblyopia (80% vs 77%; P = .372) and strabismus (89% vs 80%; P = .002) themselves. Fewer optometrists than ophthalmologists dilated eyes routinely during the first evaluation of a preschool-aged child (39% vs 93%; P < .001). The leading barrier to care for preschool-aged children reported by both optometrists and ophthalmologists was difficulty of the examination (25% vs 23%; P = .501). Optometrists reported that most of their patients were referred from community-based screening programs or by parent self-referral. Ophthalmologists reported that most of their preschool-aged patients were referred from primary care providers. CONCLUSIONS: There are different sources of referrals for optometrists and ophthalmologists. Although most eye care specialists treat amblyopia, the types of care offered by optometrists and ophthalmologists differ. Future studies should address the impact that these patterns have on outcomes and cost. The results of these studies should be shared with those responsible for screening. PMID- 14560836 TI - Long-term results of unilateral lateral rectus recession in intermittent exotropia. AB - PURPOSE: Unilateral lateral rectus recession is an alternative procedure in the treatment of moderate angle intermittent exotropia. However, long-term results of unilateral lateral rectus recession have not yet been studied. The aim of this study was to determine the long-term stability of alignment following unilateral lateral rectus recession in patients with intermittent exotropia with a deviation of 25 to 30 PD. PATIENTS AND METHODS: Thirty-two patients with intermittent exotropia with a deviation of 25 to 30 PD were included in this study. All patients underwent 8-mm unilateral lateral rectus recession by conventional techniques. RESULTS: Satisfactory results (+/- 5 PD of orthophoria) were obtained in 77.7% of the patients after a 3-year follow-up period. The mean correction achieved was 22.9 PD. CONCLUSION: Our results indicate that 8-mm unilateral lateral rectus recession is an attractive alternative procedure in the treatment of intermittent exotropia with a deviation of 25 to 30 PD. PMID- 14560837 TI - Incidence of stereopsis after treatment of infantile esotropia with botulinum toxin A. AB - PURPOSE: The incidence of stereoscopic vision following surgery for infantile esotropia has been reported, but not from patients treated with simultaneous bimedial injection of botulinum toxin A. We previously reported other findings from 41 patients treated with botulinum toxin A before the age of 12 months. The purpose of this study was to report the incidence of stereopsis in these patients after long-term follow-up. PATIENTS AND METHODS: Data were acquired by recording the response to subjective stereoscopic testing during the postinjection follow up period when reliable subjective responses could be obtained. At least 5 years had passed following injection, and the mean age of the patients was 8 years. RESULTS: Of the 41 patients, 11 were lost to follow-up, 10 had no stereopsis, 9 had gross stereopsis (according to Titmus fly tests), 1 had 800 seconds of stereoscopic vision, 2 had 400 seconds, 2 had 200 seconds, 1 had 60 seconds, and 5 had 40 seconds. CONCLUSIONS: In this study, two-thirds of the patients acquired stereopsis following simultaneous bimedial injection of botulinum toxin A for infantile esotropia. Stereopsis acquisition following injection of botulinum toxin A had a success rate comparable with that reported for surgically corrected infantile esotropia. Bimedial injection of botulinum toxin A accompanied by alternate patching prior to injection and diligent attention at ages ranging from 4 to 12 months is a simple and efficacious method for the management of infantile esotropia. PMID- 14560838 TI - Characteristics of Lisch nodules in patients with neurofibromatosis type 1. AB - PURPOSE: To assess the prevalence and location of Lisch nodules among patients with neurofibromatosis type 1. PATIENTS AND METHODS: This was a prospective observational case series in which 369 consecutive patients from a neurofibromatosis clinic participated. Predetermined inclusion and exclusion criteria were applied to these participants. The presence of Lisch nodules was determined by one of the authors (SMC). The percentage of patients with neurofibromatosis type 1 with Lisch nodules was calculated. Lisch nodules were characterized as being distributed superiorly, inferiorly, or equally in each eye. The percentage of eyes in each group was ascertained. RESULTS: Among patients of all ages with neurofibromatosis type 1, 63.2% had Lisch nodules, whereas 92% of postpubertal patients had Lisch nodules. Among eyes with Lisch nodules, 80% had an inferior distribution. CONCLUSIONS: This study corroborated previously reported data regarding the prevalence of Lisch nodules in patients with neurofibromatosis type 1 and found that Lisch nodules were predominantly located inferiorly on the iris. The authors propose that exposure to sunlight may be the mechanism for this distribution of hamartomatous nodules. PMID- 14560839 TI - Retinal features in Seckel's syndrome. PMID- 14560840 TI - Conjunctival cat-scratch disease simulating rhabdomyosarcoma. PMID- 14560841 TI - A new treatment modality for phthiriasis palpebrarum. PMID- 14560842 TI - Non-African Burkitt's lymphoma manifesting at the jaw and as a right orbital mass in a child. PMID- 14560843 TI - Silent sinus syndrome as a cause of diplopia in a child. PMID- 14560844 TI - Primary intraocular hydatid cyst with glaucoma. PMID- 14560845 TI - Diffusion tensor imaging in patients with major cerebral artery occlusive disease. AB - Diffusion tensor (DT) imaging provides quantitative information about the magnitude and the directionality (anisotropy) of water diffusion in vivo and can detect pathologic changes in brain ischemia. This study tried to detect ischemic brain damage using DT imaging in patients with symptomatic chronic major cerebral artery occlusive disease. DT imaging was performed using a 3.0 Tesla magnetic resonance (MR) scanner in 50 patients with unilateral internal carotid artery or middle cerebral artery stenosis or occlusion, who had no obvious infarct lesions on conventional MR imaging. Thirty-three patients underwent DT imaging before and after vascular reconstruction surgery. Fractional anisotropy (FA) was calculated in the middle cerebral artery territory. Preoperative FA values in the ipsilateral side were significantly lower than those in the contralateral side. After surgery, the FA value was significantly increased. DT imaging may indicate ischemic brain damage, not visualized by conventional MR imaging, in patients with major cerebral artery occlusive disease. PMID- 14560846 TI - Operative anatomy and classification of the sylvian veins for the distal transsylvian approach. AB - Methods for preservation of the sylvian veins in the transsylvian approach have not been established because of the considerable variations. This study attempted to classify the sylvian veins to facilitate systematic dissection of the sylvian fissure for sylvian veins to be preserved. The operative anatomy of the sylvian vein was examined in 82 hemispheres. The type of drainage and the pattern of branching were investigated. The superficial sylvian vein (SSV) was classified into three types according to the number of stems draining into the dural sinus on the inner surface of the sphenoid bone: The SSV was absent or hypoplastic in eight cases, the SSV was single in 38 cases, and the SSV was double in 36 cases. The SSV drained into neither the sphenoparietal sinus nor the cavernous sinus in nine cases. An anastomosis between the SSV and the deep middle cerebral vein (DMCV) was observed in 42 cases. The frontobasal bridging vein (FBBV) drained into the sphenoparietal sinus in 47 cases. The type of connection was further subdivided into four types according to the connections with the DMCV and FBBV. The venous anatomy of sylvian fissure indicates that dissection (skeletonization) of the main stem of sylvian veins from the temporal lobe should be performed to preserve the tributaries from the frontal lobe. PMID- 14560847 TI - Familial association of basilar bifurcation aneurysm and moyamoya disease--four case reports. AB - Four patients presented with familial intracranial aneurysms and familial moyamoya disease, including one patient with both familial intracranial aneurysm and moyamoya disease. Basilar bifurcation aneurysms were present in two patients, moyamoya disease in one, and both basilar bifurcation aneurysm and moyamoya disease in one. These events are most likely to arise from different genetic abnormalities associated with basilar bifurcation aneurysm and moyamoya disease. PMID- 14560848 TI - Serial neuroimaging of encapsulated chronic intracerebral hematoma with repeated hemorrhage--case report. AB - A 69-year-old woman presented with headache and short memory disturbance. Computed tomography (CT) demonstrated a small cystic mass lesion in the left temporal lobe. CT and magnetic resonance imaging showed that this lesion enlarged with repeated hemorrhages, associated with progressive amnesia and headache during 3 years follow up. Surgery demonstrated a well-demarcated hard mass lesion in the medial temporal lobe through a transcortical approach after opening left sylvian fissure. The lesion was located entirely in the brain parenchyma and was removed en-bloc after cutting some capillary-like vessels on the capsule. The histological diagnosis was encapsulated old hematoma. The histological findings suggested that expansion of the lesion was due to multiple bleedings from the sinusoidal vessels in the capsule fed by small feeding arteries. The mass effect due to the expansion of the encapsulated hematoma caused progressive short-term amnesia and headache, which were completely resolved by the surgical removal. PMID- 14560849 TI - Pseudoaneurysm formation at the rupture site of a middle cerebral artery aneurysm -case report. AB - A 42-year-old man suffered subarachnoid hemorrhage manifesting as sudden severe headache one month before admission. On admission, his headache had subsided and he had no neurological deficits. Cerebral angiography demonstrated an aneurysm originating from the bifurcation of the right middle cerebral artery. The aneurysm was irregular, with a snowman-like shape. Neck clipping of the aneurysm was carried out through a right pterional approach. Intraoperatively, a red, pulsating sac mimicking a blood clot (the snowman's "head") was located over the yellowish, thick-walled portion of the aneurysm. Exploration around the aneurysm detached the red sac from the thick-walled portion of the aneurysm. There was a small tear in the apex of the thick-walled aneurysm sac. A Sugita clip was applied to the neck of the true aneurysm. The postoperative course was uneventful and he was discharged one month later without neurological deficits. Histological examination of the red-colored sac showed the features of pseudoaneurysm. The red sac may have been a pseudoaneurysm covering the rupture site of the true aneurysm. PMID- 14560850 TI - Central nervous system lesions associated with hereditary hemorrhagic telangiectasia--three case reports. AB - Hereditary hemorrhagic telangiectasia (HHT) is easily overlooked in patients with central nervous system (CNS) lesions. Our clinical experience of three patients with CNS lesions associated with HHT stresses the importance of considering HHT. A 23-year-old male presented with consciousness disturbance and right hemiparesis. Emergency cerebral digital subtraction angiography revealed occlusion of the left middle cerebral artery and the left anterior cerebral artery. Pulmonary arteriography showed three pulmonary arteriovenous malformations (AVMs). A 62-year-old male presented with consciousness disturbance and sensory aphasia. Magnetic resonance imaging revealed a ring-enhanced lesion in the left temporal lobe which was removed by left frontotemporal craniotomy. The diagnosis was brain abscess. Chest computed tomography (CT) revealed two pulmonary AVMs. A 32-year-old female presented with progressive mild weakness in her left hand. Initial CT showed subcortical hemorrhage in the right frontal lobe. Cerebral angiography revealed no vascular malformations, but chest CT disclosed five pulmonary AVMs. All three patients had a family history of HHT. The possibility of HHT is important to consider in patients with cerebrovascular disease (CVD) or brain abscess to prevent complications, not only in the patients but also their blood relatives. Therefore, the medical and family history of patients with CVD or brain abscess should be investigated and HHT should be considered during the physical examination. PMID- 14560851 TI - Glioblastoma with metastasis to the spleen--case report. AB - A 47-year-old woman presented with headache and left homonymous hemianopsia. T1 weighted magnetic resonance (MR) imaging with contrast medium showed a mass lesion with ring-like enhancement in the right temporo-occipital lobe. The patient underwent surgery, focal irradiation, and chemotherapy. The histological diagnosis was glioblastoma. Four months after the operation, the patient again developed headache and left homonymous hemianopsia in addition to vomiting and mild left hemiparesis. MR imaging showed recurrence of the tumor and hydrocephalus. The patient underwent a second craniotomy and placement of a ventriculoperitoneal shunt. Intraoperative findings revealed that the transverse sigmoid sinus was occluded by tumor invasion. The patient died of intraventricular dissemination 2 months after the second operation. Autopsy revealed metastases in the spleen and lungs. Glioblastoma with metastases to the spleen is very rare. The prognosis for patients is poor. Excessive therapy should not be used for patients with extracranial metastases from glioblastoma. PMID- 14560852 TI - Subthalamic nucleus tumor causing hyperphagia--case report. AB - A 42-year-old woman with locally advanced breast cancer developed headache just after completing adjuvant chemotherapy. Magnetic resonance imaging revealed a mass located in the left subthalamic nucleus (STN) and involving the posterior part of the thalamus and the hypothalamus. The patient refused a radiologically guided biopsy and gamma knife treatment was not financially possible. Palliative whole brain radiotherapy with hormonal therapy was administered. The patient gained 19 kg body weight during 4 months follow up because of hyperphagia. This solitary tumor, either a breast cancer metastasis or a primary tumor, involving the STN is extremely unusual. PMID- 14560853 TI - Discrepancy between [18F]fluorodeoxyglucose and 11C-methionine positron emission tomography findings in Sturge-Weber syndrome--case report. AB - Cerebral blood flow and metabolism were evaluated in an adult with symptomatic intractable epilepsy and Sturge-Weber syndrome (SWS) manifesting as angiomas in the left cerebral hemisphere. 99mTc-ethylcysteinate dimer single photon emission computed tomography detected reduced blood flow in the entire left cerebral hemisphere, and [18F]fluorodeoxyglucose positron emission tomography (PET) showed decreased glucose metabolism in the left cerebral hemisphere. These findings indicated hypofunction of the left cerebral hemisphere, which caused the right hemiparesis. 11C-methionine (11C-Met) PET revealed high 11C-Met accumulation in the angiomas in the left cerebral hemisphere. Immunostaining for glial fibrillary acidic protein showed positive reaction in the lesions. Gliosis is a likely mechanism for the 11C-Met accumulation, which is possibly associated with progressive calcification in the angiomas and retarded growth of patients with SWS occurring over many years. PMID- 14560854 TI - Open median nerve release using double mini skin incision in patients with carpal tunnel syndrome: technique and clinical results. AB - Patients who undergo open carpal tunnel surgery frequently complain of the postoperative cosmetic appearance at the site of the incision on the palm. This problem occurs as a result of excessive scar formation, and the long incision. A double mini skin incision, each 1 cm long, was used in the surgical treatment of carpal tunnel syndrome. The transverse carpal ligament was easily sectioned. Postoperatively healing was good with no excessive scar formation. PMID- 14560855 TI - Supplementary comment on "Computed tomography evaluation of air cells in the petrous bone--relationship with postoperative cerebrospinal fluid rhinorrhea". PMID- 14560856 TI - Management of the severely traumatised limb. PMID- 14560857 TI - Biostatistics 101: data presentation. PMID- 14560858 TI - Skeletal union following long bone reconstruction using vascularised fibula graft. AB - Thirteen patients had skeletal reconstruction using vascularised fibula graft following resection of the diseased bone. Eleven patients had reconstruction of the lower limbs and two patients of the upper limbs. Clinical and radiographical evidence union were achieved with the average time of 32 weeks (earliest eight weeks). Six out of 11 patients (54%) in lower limb reconstruction started weight bearing at the average of 27 weeks. Bony union in this study is comparable with other studies using vascularised fibula graft. PMID- 14560859 TI - Initial experience of laparoscopic management of apparent early endometrial cancer. AB - The traditional treatment for endometrial cancer is a staging laparotomy. In patients with apparent early stage disease, laparoscopic assisted vaginal hysterectomy has been employed as an alternative. We present a retrospective uncontrolled case series of 16 cases with apparent stage I endometrial cancer from November 1994 to April 2001 managed by laparoscopic assisted surgical staging (LASS) at the Gynaecological Oncology Department, KK Women's and Children's Hospital, Singapore. We have analysed outcome measures in terms of intra-operative complications, surgical morbidity and length of hospital stay, and have evaluated treatment success in terms of conversion to laparotomy, recurrent disease and mortality. Our findings indicate that LASS is a viable option in selected patients with apparent early endometrial cancer. Our initial experience with this modality of treatment thus proves that it has great potential in both treatment success and reduction of surgical morbidity. A larger case controlled trial would be ideal in order to substantiate these benefits. PMID- 14560860 TI - Ultrasonography is not reliable in diagnosing liver cirrhosis in clinical practice. AB - BACKGROUND: The diagnosis of liver cirrhosis is important in the evaluation and management of patients. Liver biopsy is the gold standard but it is invasive. Ultrasonography is a non-invasive and useful modality in assessing the liver for certain conditions but its sensitivity and specificity in diagnosing cirrhosis is unknown locally. AIM: To assess the accuracy of ultrasonography in diagnosing compensated liver cirrhosis in daily clinical practice outside the context of clinical trials. METHODS: All the liver biopsies were identified from the Pathology Logbook retrospectively from January 1998 to March 2001. Only patients who had both liver biopsy and ultrasonography with no clinical evidence of cirrhosis were included. Patients with incomplete data, hepatoma or liver secondaries were excluded. Ultrasonographic diagnosis of cirrhosis was based on nodularity or irregularity of the liver surface, small liver size, coarse echotexture and increase attenuation by using the 3.5 to 5 MHz transducers. RESULTS: A total of 151 liver biopsies were performed during this period. Eighty eight patients who had both ultrasound and liver biopsy were analysed. Seventeen patients had ultrasonographic diagnosis of cirrhosis but only six cases were proven by a liver biopsy. On the other hand, 10/16 cases of biopsy-proven cirrhosis were "missed" by ultrasound. Thus, the sensitivity of ultrasonography in diagnosing cirrhosis was 37.5% and the specificity was 84.7%. The positive and negative predictive values were 35.3% and 85.9% respectively. CONCLUSION: Low frequency ultrasonography is not a sensitive test for the diagnosis of liver cirrhosis in daily clinical practice. PMID- 14560861 TI - Recovery following tonsillectomy a comparison between tramadol and morphine for analgesia. AB - BACKGROUND: The optimal method of intraoperative analgesia for adult tonsillectomy is uncertain. It is important that recovery should be rapid so that the airway is not compromised. Tramadol hydrochloride is an analgesic with mixed mu and non-opioid activities which has less respiratory depression effects compared to morphine. PATIENTS AND METHODS: We compared the recovery characteristics of patients undergoing tonsillectomy after they were given either morphine or tramadol for intra-operative analgesia. Seventy-nine ASA (American Society of Anesthesiologists) I patients were randomised to receive either tramadol 1.5 mg/kg (n = 44) or morphine 0.1 mg/kg (n = 35). A standard propofol desflurane based general anaesthetic technique was used. RESULTS: Patients given tramadol recovered faster compared to morphine as demonstrated by the earlier eye opening at reversal (mean +/- SD, 4.7 +/- 1.5 min versus 5.6 +/- 1.8 min, p = 0.04). There was also significantly less nausea and vomiting in the patients given tramadol as compared to those given morphine (6.8% versus 28.6%, p = 0.01). There were no other clinically important adverse effects in either group. CONCLUSION: We conclude that tramadol given for intra-operative analgesia during tonsillectomy results in faster recovery with significantly less nausea and vomiting in the early postoperative period. PMID- 14560862 TI - Percutaneous drainage of liver abscess complicated by hepato-venous fistula. AB - Severe systemic sepsis after percutaneous drainage of liver abscess is rare. We report two cases of hepato-venous fistulas between hepatic abscesses and hepatic/portal veins documented on abscessography during percutaneous drainage of liver abscesses, which resulted in severe sepsis and a stormy post drainage clinical course. Liver abscesses can rupture into the portal and hepatic veins causing worsening of systemic sepsis especially when they are in close proximity to each other. During percutaneous drainage, care must also be taken to avoid overinjection of the abscess, which can worsen the fistula. The ensuing sepsis is severe and requires aggressive intensive medical care and ventilatory support to tide the patient over the septic episode. PMID- 14560863 TI - Metastatic carcinoma of the cervix presenting as a psoas abscess in an HIV negative woman. AB - A case of advanced carcinoma of the cervix presenting as a psoas abscess is described. This unusual presentation has been reported in association with AIDS but is extremely rare in HIV-negative women. This case report emphasises the importance of awareness of unusual causes of the psoas abscess in an era when the classical tuberculous abscess is fast disappearing. PMID- 14560865 TI - Liver transplantation for hepatocellular carcinoma: how far can we push the envelope? PMID- 14560864 TI - Giant cell arteritis (temporal arteritis, cranial arteritis) and a case from Singapore. AB - Giant cell arteritis as the underlying cause of blindness in the elderly is common in the West but is not seen except on rare occasions in South East Asia. We describe an 86-year-old Chinese man from Singapore who presented with a central retinal artery occlusion. Biopsy of a prominent superficial temporal artery established the underlying cause to be giant cell arteritis which was also the eventual cause of death as the condition resulted in rupture of a dissecting aneurysm of the aorta. Giant cell arteritis should be considered in all cases of ischaemic eye disease in the elderly. The importance of early diagnosis lies in the very high incidence of second eye involvement within days or at most weeks in untreated patients. A high index of suspicion is required for diagnosis of this condition which is likely under-diagnosed in our local context. PMID- 14560866 TI - Imaging and the child with abdominal pain. AB - Abdominal pain is a common presenting symptom in children. A substantial percent of unscheduled paediatric office visits and paediatric emergency room visits are due to abdominal pain. Children may present with an acute episode of pain or with chronic, recurrent pain. Only a small fraction of children presenting with abdominal pain will prove to have an organic cause necessitating interventional management. In this essay, a discussion of abdominal pain is presented, with particular emphasis on the role of radiologic imaging tests to investigate potential causes of abdominal pain. The more common and clinically significant causes of abdominal pain in a child will be discussed in greater detail. PMID- 14560867 TI - Article: Intra-arterial embolisation in otolaryngology--a four-year review. PMID- 14560868 TI - Clinics in diagnostic imaging (86). Ruptured bronchogenic cyst. AB - An eight-year-old boy presented with a sudden onset of chest pain. He had been diagnosed to have a left spontaneous pneumothorax. Chest radiographs and computed tomography of the chest showed a thin-walled cyst in the left lower lobe. Thoracotomy and a segmentectomy of the apical segment of the lower lobe was performed, confirming the diagnosis of a ruptured bronchogenic cyst. Imaging findings of various pulmonary cystic lesions in children are discussed. PMID- 14560869 TI - Protecting the public: myth or reality? PMID- 14560870 TI - California, U.S. show no improvement on report card. PMID- 14560871 TI - The pathophysiology, medical management, and dental implications of children and young adults having attention-deficit hyperactivity disorder. AB - Attention-deficit hyperactivity disorder is considered to be the most common neurobehavioral disorder among school-aged children. Children who have ADHD exhibit symptoms of inattention, hyperactivity, and impulsivity that are extreme for their age. These symptoms adversely affect academic performance, vocational success, and social-emotional development. The disorder is usually treated with stimulant medications, but on occasion antidepressants and certain antihypertensive agents are used. Dentists caring for individuals with ADHD must be familiar with the manifestations of the disease and its associated features so they can garner the maximum level of cooperation from the patient. They must also be familiar with the drugs used to treat the disorder because many of these pharmaceuticals cause adverse orofacial and systemic reactions and may interact with dental therapeutic agents. PMID- 14560872 TI - The pathophysiology, medical management, and dental implications of autism. AB - Autism is a lifelong, severe, developmental disorder that appears initially in infancy and early childhood and impairs the acquisition of some of the most important skills in human life. The disease is characterized by impaired social interactions, verbal and nonverbal communication deficiencies, limited activities and interest, and repetitive behaviors. Often accompanying the disorder are behavioral disturbances, such as self-mutilation and aggression, psychiatric symptoms, and seizures, which necessitate the administration of multiple medications to help the affected individual participate effectively in the educational and rehabilitative process. Dentists caring for these people must be familiar with the manifestations of the disease and its associated features so that they can garner the maximum level of cooperation. They must also be familiar with the medications used to treat the associated features of the disorder because many of these pharmaceuticals cause untoward orofacial and systemic reactions and may precipitate adverse interactions with dental therapeutic agents. PMID- 14560873 TI - The pathophysiology, medical management and dental implications of fragile X, Rett, and Prader-Willi syndromes. AB - Fragile X, Rett, and Prader-Willi syndromes are a group of inherited disorders that often present with varying degrees of mental retardation and challenging behaviors. Dentists caring for individuals with these disorders must be familiar with the manifestations of these diseases and their associated features so they can garner the maximum level of cooperation from the patient. They must also be familiar with the medications (anticonvulsants, antihypertensives, antidepressants, antipsychotics, and central nervous system stimulants) used to treat the associated behaviors, because many of these pharmaceuticals cause clinically evident orofacial and systemic reactions, and may precipitate adverse interactions with dental therapeutic agents. PMID- 14560874 TI - Approaching the pediatric dental patient: a review of nonpharmacologic behavior management strategies. AB - One of the difficulties of the practice of dentistry is being responsible for the highest quality of care for patients while often having to propose alternate solutions for a variety of reasons, which include financial concerns. Dental practitioners treating young children have the added responsibility of gaining their patient's cooperation to render the best treatment. Determinants that influence the development of a behavioral strategy for a young patient include disease status, the child's physical and mental development, parental characteristics, and provider personality and capabilities. Classic strategies- including a supportive office environment, "tell-show-do," successive approximation, distraction, behavior shaping, and retraining--must be matched to the characteristics of each child and family situation. Current cultural trends suggest that disciplinary forms of behavior management strategies--such as hand over-mouth, physical restraint, and even voice control--are losing societal acceptance. PMID- 14560875 TI - What fools these mortals be. PMID- 14560876 TI - The social reality of the imaginary audience: a grounded theory approach. AB - Traditional approaches to understanding the imaginary audience are challenged in this study. Three hundred sixty-one British schoolchildren (aged 14 and 15 years) were asked to express their worries and concerns, using grounded theory methodology. Qualitative responses were collated and coded according to emerging categories, with "what other people think" identified as the central concern. In particular, the findings are used to critique Elkind's (1967) theory of adolescent egocentrism. Data presented in this study suggest that adolescents worry about what other people think because there are real personal and social consequences. Such concerns are seen as being based in social reality and are not imaginary as Elkind suggested. In conclusion, new methodologies which place young people at the center of the analysis are advocated. PMID- 14560877 TI - "You learn and learn and learn.... and then you are an adult": parental perceptions of adolescence in contemporary Swaziland. AB - The developmental period of adolescence is explored in Swaziland from a multidisciplinary perspective. The study compares early anthropological research with contemporary interviews of Swazi parents. While the Swazi language, siSwati, does not have a term for "adolescent," there is evidence of a definite developmental period which could be defined as adolescence. Parents view this period as being qualitatively different from childhood and adulthood, but they also have gender-related interpretations of its nature and length. While biological maturation (especially menarche for females) is viewed as a vital step in becoming an adult, behaviors must be learned during this time (i.e., "adolescence") before one is completely of adult status. PMID- 14560878 TI - Impact of exposure to community violence and psychological symptoms on college performance among students of color. AB - This study examined longitudinal relationships among exposure to chronic community violence during high school, psychological distress during the first semester of college, and academic performance during the first three semesters of college. The sample comprised 385 students of color in a large city. Exposure to community violence and psychological distress were measured with additive scales; academic performance (school persistence, grade point average) was obtained from transcripts. It was found that exposure to community violence and academic performance were not related; exposure to community violence and psychological distress were related; psychological distress and college persistence were related; and psychological distress and grade point average were not related. The findings are consistent with the causal chain model; specifically, that the effects of exposure to community violence in high school on academic performance in college are mediated by psychological distress. PMID- 14560879 TI - Incarcerated adolescent girls: personality, social competence, and delinquency. AB - The aim of this study was to gain greater insight into the relationship of delinquency to personality and social competence in incarcerated girls. It was hypothesized that agreeableness and conscientiousness would correlate negatively with delinquency, and that the sensation-seeking aspect of extroversion would correlate positively. Two theoretical views were used to predict the relationship between social competence and delinquency: (a) delinquency as an expression of lack of social competence (i.e., social deficit), and (b) delinquency as socially competent and rationally analyzed behavior. The first view implies a negative correlation between delinquency and social competence, while the second indicates a positive one. The participants were thirty-three 12- to 18-year-old incarcerated girls. An adaptation of the Self-Reported Delinquency Scale was used for measuring delinquency; personality was measured using the Five-Factor Personality Inventory; and the Scale for Interpersonal Behavior was used to measure social competence. Agreeableness was not found to correlate with the overall delinquency score or with any specific type of delinquency or crime. The more crimes the girls reported, the less conscientious they were, and the more neurotic and open they were. The correlation between delinquency and extroversion was not statistically significant. A higher level of delinquency was accompanied by higher social competence when negative self-assertion was required (i.e., a low level of tension was felt in expressing negative feelings that were appropriate to the situation). In other social situations, however, the girls reported more situation-inappropriate feelings of tension. It was concluded that some personality factors and aspects of social competence can predict delinquency. PMID- 14560880 TI - The relationship between the living arrangements of university students and their identity development. AB - One of the developmental challenges facing the emerging adult is to live independently of his/her family of origin. Research has shown that the living arrangements of late adolescents/young adults also affect aspects of their personal development. This study examined the relationship between university students' living arrangements, their identity development status, the degree of life difficulties experienced, and the manner in which they coped with their difficulties. In general, it was found that those who did not reside in their parents' home had greater levels of problems but also used more direct, problem focused coping strategies than did those who resided at home. Individuals residing away from their parents were also more likely to have established an adult identity, whereas those who resided with their parents were more likely to be still in the process of developing an adult identity. PMID- 14560881 TI - Salient worries of at-risk youth: needs assessment using the things I worry about scale. AB - At-risk students in the southern United States were surveyed using the Things I Worry About Scale. From their responses, 13 categories were ranked from most salient to least salient. The rank order obtained in the present study was compared to that obtained with a normative group of youths in Northern Ireland. The two rank orders were found to be significantly correlated. The relatedness of the rank orderings of the two seemingly disparate samples suggests the universality of the Things I Worry About Scale. The merits of using the scale as part of a comprehensive needs assessment are discussed. PMID- 14560882 TI - Dissociative experiences and anger proneness in late adolescent females with different attachment styles. AB - The study examined the relationships among dissociative experiences, anger proneness, and attachment styles in late adolescent females. One hundred sixty two college students (mean age = 17.5 years) were assessed using the Dissociative Experience Scale (DES), the State-Trait Anger Expression Inventory (STAXI), and the Adult Attachment Questionnaire (AAQ), a self-report tool for assessing attachment styles (avoidant, secure, and ambivalent-resistant) in close relationships of youths and adults. Significant correlations were obtained between DES scores (total and factorial) and STAXI scores (State Anger, Trait Anger, Anger/In, and Anger/Expression), confirming in a nonclinical sample the connection between anger proneness and dissociation described in patients with dissociative disorders. Insecure females, particularly ambivalent ones, scored higher on the DES, supporting van der Kolk's hypothesis of an inverse relationship between secure attachment and dissociative tendency. Moreover, insecurely attached females showed more anger proneness, with some differences between ambivalent and avoidant types. Further research should be conducted to examine these relationships in males, as well as to clarify the role of insecure attachment in anger management and the recourse to dissociation in late adolescence as a protective response to trauma and emotional distress. PMID- 14560884 TI - Coping assessment in adolescents. AB - The aim of this work was to obtain an abridged form of the Adolescent Coping Scale (Spanish version) that included items representative of the various factorial dimensions identified as stable. The scale (80 items) was administered to 800 Argentine adolescents. Factor analyses, principal axes and oblimin solution, were performed. An abridged scale was obtained, consisting of 46 items that were grouped into 11 dimensions: Cognitive Redefinition, Self-Blame, Fatalism, Evasion Through Amusement, Problem-Focused Coping (which includes requests for information and action), Evasion Through Physical Activity, Emotional Support, Emotional Discharge and Somatization, Anxiety, Isolation, and No Action. The reliability coefficients were found to be satisfactory. PMID- 14560883 TI - Feeling at home in one's school: a first look at a new measure. AB - We report on the development of the Hospitality Scale, which measures two aspects of adolescents' perception of social capital in school-the extent to which they perceive that they have social capital and the extent to which they provide social capital to others. The scale was developed in reference to the literature exploring adolescent social isolation and social tolerance, as well as constructs developed in research on adults, especially related to psychological sense of community and collective efficacy. We examine the reliability of the scale and the association between scale scores and behavioral factors that may reflect social isolation. PMID- 14560885 TI - Dream content of schizophrenic, nonschizophrenic mentally ill, and community control adolescents. AB - This study compared the manifest dream content of 20 schizophrenic adolescent inpatients whose medications were stable for at least four weeks, 21 adolescent inpatients with other mental disorders (nonschizophrenic group) matched for age and gender, and 31 matched community controls. All participants were administered the standardized Formal Dream Content Rating Scale (FDCRS), which evaluates dream related anxiety, cognitive disturbance, implausibility, involvement, primitivity, and recall, as well as two additional scales measuring emotional expression and duration of dream report. The Positive and Negative Symptoms Scale (PANSS) was administered to the two inpatient groups. The community controls demonstrated more involvement and emotional expression than the schizophrenic patients; furthermore, they demonstrated more implausibility and had a greater duration of dream report compared with the nonschizophrenic group. In the schizophrenic patients only, elevated scores on the negative subscale of the PANSS were significantly correlated with lower scores on involvement, emotional expression, and dream recall. No relationship was found between the positive subscale of the PANSS and any of the FDCRS subscales. These results suggest that psychopathology per se, rather than the specific psychiatric disturbance, may be associated with impoverishment of dream content, and that negative, rather than positive, schizophrenic symptomatology may be influential in the dream content of schizophrenic youngsters. PMID- 14560886 TI - Medical decision-making and minors: issues of consent and assent. AB - The physician-patient relationship has been described as an egalitarian partnership in which patients and physicians work together to make healthcare decisions. Although adults receive considerable encouragement to become active participants in medical decision-making, children and adolescents often have little voice in such decisions and are granted limited access to confidential medical care. After a brief discussion of legal perspectives on informed consent, the present review examines the developmental literature on children and adolescents' capacities to make medical decisions that are informed, voluntary, and rational. The purposes and benefits of assent are identified. Remaining questions of how to evaluate capacity and balance parental and minor autonomy are explored. PMID- 14560887 TI - Expressed attitudes of adolescents toward marriage and family life. AB - In the U.S., modifications in family structure and in attitudes concerning marriage and family life have been numerous. Areas such as sexual behavior and alternative living arrangements have become highly varied and nontraditional compared to past generations. This study examined the attitudes of adolescents toward aspects of marriage and family life. The majority of adolescents expressed negative attitudes toward divorce and viewed marriage as a lifelong commitment. While only about a third of the adolescents expressed positive attitudes toward premarital sex, a majority indicated they would engage in sexual intercourse before marriage, or already have. Interestingly, about half of the adolescents held positive attitudes toward cohabitation. Lastly, the adolescents demonstrated a growing acceptance of premarital counseling and psychoeducational interventions regarding marriage and family life. PMID- 14560888 TI - Jamaican child-rearing practices: the role of corporal punishment. AB - The family is the most prominent social group that exists. It prepares its members for the various roles they will perform in society. Yet, the literature has unequivocally singled out the family as the most violent social group, with parental violence against children being the most prevalent type of family violence. While societies like the United States, Japan, and Sweden have taken a hard line on physical punishment and shifted to a gentler approach to discipline, harsh disciplining of children persists elsewhere. In the Caribbean, and Jamaica in particular, child-rearing and disciplinary practices that would warrant child abuse charges in other Western societies are rampant. This article examines the child-rearing techniques of Jamaican adults and their assumed effects on child outcomes. It also examines the plausibility of the assumption that the harsh physical punishment meted out to children is partially responsible for the current social problems of that island nation. We recommend approaches to tackle the broad goals of addressing familial and societal practices that compromise children's development and well-being. PMID- 14560889 TI - Central causes of hypogonadism--functional and organic. AB - Whether caused by environmental factors, lesions, genetic mutations, drug interactions, or unknown origins, the path of the central causes of hypogonadism frequently leads back to the GnRH pulse generator. In some cases, the cause can be unequivocally traced to a single factor, such as some of the congenital syndromes previously described. In most instances, however, hypogonadism is occult or functional. Because of the wide spectrum and complexity of underlying causes, a definitive diagnosis, especially in functional causes of the disorder, is not always attainable. PMID- 14560890 TI - Mechanisms of follicular dysfunction in 46,XX spontaneous premature ovarian failure. AB - Approximately one half of young women who have 46,XX spontaneous premature ovarian failure have ovarian follicles remaining in the ovary. These follicles function intermittently and unpredictably, and pregnancies can occur in these women without intervention, even many years after the diagnosis. Clearly, the term "premature menopause" is an inaccurate term for this condition. At present, there are no proven therapies that will improve follicular function for these women. Inappropriate luteinization related to low follicle number seems to be a major mechanism of follicular dysfunction. Autoimmune oophoritis, although apparently an uncommon cause of follicular dysfunction in these women, nonetheless presents the opportunity to develop an effective therapy to restore fertility. Young women with 46,XX spontaneous premature ovarian failure benefit from the care of a sensitive clinician, one who is willing to spend a little more time informing them about the diagnosis and referring them to other sources of information. With appropriate medical management and emotional support provided by a sensitive clinician, most young women with 46,XX spontaneous premature ovarian failure will lead happy, healthy, and fulfilling lives. PMID- 14560891 TI - Assessment and management of anovulatory infertility in polycystic ovary syndrome. AB - PCOS is the most common cause of anovulatory infertility. Anovulation in PCOS is exacerbated by weight gain and improved by calorie restriction in overweight subjects. Fertility can usually be restored by appropriate choice of induction of ovulation, but careful monitoring is required, even when using clomiphene alone. PMID- 14560892 TI - An evidence-based evaluation of endometriosis-associated infertility. AB - Although endometriosis is associated with infertility, a clear causal relationship has yet to be established, unless adhesive disease is found. Despite this indirect association, multiple theories have been promulgated and studies are currently underway to investigate theoretic pathogenetic mechanisms. The data regarding the treatment of endometriosis-associated infertility are limited and conflicting; however, some general preliminary conclusions can be drawn. It seems that, with early-stage disease, surgical treatment increases pregnancy rates. Using the US Preventive Services Task Force classification scheme, the evidence in support of this finding is of the highest quality, or level I. Surgical treatment for moderate and severe disease also confers benefit, although the evidence in support of this treatment is of lesser quality, level II-3 by the scheme. Medical treatment, particularly if it induces an anovulatory state, has no benefit and may delay fertility. This evidence is again of the highest quality, with a classification of level I. Although assisted reproductive technologies are of benefit regarding fertility for women with endometriosis, the IVF evidence is inconclusive, with both treatments being evaluated by at least one randomized, controlled trial conferring a level I classification to the evidence. It is unclear at this time whether endometriomas have an impact on IVF outcome. The evidence consists of only a few lower-quality studies, with a classification level of II-2. Despite the haziness of current insight into the treatment of endometriosis-associated infertility, well-designed clinical trials and basic mechanistic investigations are underway in many reproductive medicine centers. As the data from these scientific inquiries emerge, clinicians will have a clearer view of effective treatment regimens for endometriosis. PMID- 14560893 TI - Age-related decline in fertility. AB - A progressive decline in fecundity with advancing age is a reality, attributed primarily to the detrimental impact of various aging processes on female gametes. Despite medical advances that have dramatically prolonged the female life span, declining numbers and deteriorating quality of oocytes, and an increasing incidence of meiotic errors and aneuploidy of gametes and embryos, reduce clinical pregnancy rates and escalate pregnancy wastage. Increased fetal aneuploidies in ongoing pregnancies and an increased predisposition to obstetric morbidities further contribute to the diminishing reproductive successes associated with advancing age. The age of male partners, despite the decline in semen parameters and sexual performance with aging, does not appear to have a major impact on the eventual fertility of the aging couple. The contributions of age-related impaired sexuality and ejaculatory problems, although slight albeit significant, to declining fertility in the aging should be appreciated in appropriate cases. With the realization of the age-related detriment on fertility potential and the limitations of available therapeutic interventions, management of subfecundity in women beyond their mid-30s should be approached aggressively. Success of ovulation induction with clomiphine citrate or gonadotropins is marginal in women aged older than 40 years; a case can be made to proceed directly with ART in women in this age group, especially when there is coexisting male factor or pelvic disease. Except for the use of donor oocytes, the outcome of various therapeutic interventions to optimize reproductive performance in women aged older than 44 years remains dismal. A broader application of PGD techniques may contribute to improved live birth rates in reproductively aging women. The greater likelihood of obstetric complications in pregnancies resulting from donor oocytes and an increased prevalence of age-related medical problems complicating pregnancy should prompt a thorough medical evaluation before proceeding with ART. PMID- 14560894 TI - Male factor infertility. AB - There is a male factor involved in up to half of all infertile couples. Potential causes of male factor infertility are many and require thorough evaluation for their accurate elucidation. A complete medical history in conjunction with a focused examination can allow for an appropriate choice of laboratory and imaging studies. The semen analysis is a crucial first step, but it is by no means sufficient to determine cause or dictate therapy. A systematic approach is necessary to help guide the evaluation and exclude less likely causes. The causes discussed within this article are broad, and the prognosis for any given couple depends, in large part, on the cause of the infertility. Without a firm understanding of the genetics, anatomy, physiology, and their interactions necessary to permit full functioning of the male reproductive system, the evaluation becomes an inefficient exercise that often fails to elucidate the precise cause of infertility. Treatment success relies not just on a clinical diagnosis but on a determination of the cause of the male factor infertility. Therefore, couples with a component of male factor infertility need a systematic evaluation directed at the male partner to maximize their reproductive potential. PMID- 14560895 TI - Evaluating the efficacy of intervention. AB - The standard for reaching a verdict in civil trials is "the preponderance of the evidence." This is a valid standard to apply to evaluating the medical literature as well. Every published report should be given weight based on physicians' judgment of its reliability. Larger-scale studies should be weighted more heavily than smaller-scale studies, randomized trials more heavily than observation studies, rigorously designed trials more heavily than studies that may be biased. If there seems to be a relationship between a factor and an outcome, this does not necessarily imply that the factor caused the outcome. The association could be a result of chance variation between individuals. Statistical testing allows researchers to exclude chance as a likely cause of the relationship, but this is the only explanation ruled out by a significance test. The relationship could be a result of bias: bias in the selection of individuals for the study, bias in measurement of the factor or the outcome, or bias in differential loss to follow up. A thorough analysis of the data is necessary to identify and exclude other possible explanations of the association. A government agency dealing with environmental regulations experimented with replacing administrative law judges with scientists. It was believed that scientists were better qualified to make the necessary technical evaluations. The experiment was not considered successful because, as one observer remarked, "Judges are used to having to reach a verdict within a short period of time based on whatever evidence is presented--scientists just can't seem to make decisions." This illustrates a similar difference between the role of the researcher and the role of the physician. The physician must decide the best treatment for each patient based only on whatever evidence is available. There is little question, however, what the patient would decide if allowed to choose between receiving the standard therapy today or waiting 5 years until conclusive scientific evidence has been obtained about whether an alternative treatment is better. Few patients would choose to wait. Yet the decisions a physician makes today will be criticized in 5 years, especially by those who never have had to make similar decisions themselves. Some decisions will turn out to be wrong when additional data are available, but physicians must make choices based on the best data currently available. The choice of medicine as a profession implies the acceptance of a life sentence to jury duty: the evidence will never stop accumulating, and the verdict must be continually reevaluated. PMID- 14560896 TI - Biomechanics and pathophysiology of flat foot. AB - When the foot works properly it is an amazing, adaptive, powerful aid during walking, running, jumping, and in locomotion up or down hill and over uneven ground. Dysfunction of the foot can often arise from the foot losing its normal structural support, thus altering is shape. An imbalance in the forces that tend to flatten the arch and those that support the arch can lead to loss of the medial longitudinal arch. An increase in the arch-flattening effects of the triceps surae or an increase in the weight of the body will tend to flatten the arch. Weakness of the muscular, ligamentous, or bony arch supporting structures will lead to collapse of the arch. The main factors that contribute to an acquired flat foot deformity are excessive tension in the triceps surae, obesity, PTT dysfunction, or ligamentous laxity in the spring ligament, plantar fascia, or other supporting plantar ligaments. Too little support for the arch or too much arch flattening effect will lead to collapse of the arch. Acquired flat foot most often arises from a combination of too much force flattening the arch in the face of too little support for the arch. Treatment of the adult acquired flat foot is often difficult. The clinician should remember the biomechanics of the normal arch and respond with a treatment that strengthens the supporting structures of the arch or weakens the arch-flattening effects on the arch. After osteotomies or certain hindfoot fusions, the role of the supporting muscles of the arch, in particular the PTT, play less of a role in supporting the arch. Rebalancing the forces that act on the arch can improve function and lessen the chance for further or subsequent development of deformity. PMID- 14560897 TI - Adult acquired flat foot deformity: clinical and radiographic examination. AB - Symptomatic adult acquired flat foot deformity is encountered in the orthopedic office on a frequent basis. Although many causes exist, a careful history and a stepwise approach to the physical examination will clue the examiner into making the correct diagnosis and provide appropriate treatment. Radiographs serve as an adjunct and assist in verifying the examination findings. CT, US, and MRI are helpful modalities for surgical planning or when the diagnosis remains questionable. PMID- 14560898 TI - The indications and biomechanical rationale for various hindfoot procedures in the treatment of posterior tibialis tendon dysfunction. AB - Several procedures are available for the treatment of posterior tibialis tendon dysfunction. The procedure that is chosen for a specific patient should address the patient's unique problem and make biomechanical sense. A fusion should be avoided, if possible; however, if a fusion is the appropriate solution, one should not hesitate to do it. PMID- 14560899 TI - A classification of severity with an analysis of causative problems related to the type of treatment. AB - The current classification systems provide broad guidelines for treating the acquired adult flat foot. They essentially place the acquired flat foot into categories of no deformity, flexible deformity, or rigid deformity, as well as the condition of the PTT; however, there are many more variables that need to be taken into account to provide optimal care. As our knowledge and experience grows in treating the adult flat foot, more options will be available. The physician treating this condition must stay up-to-date and must not rely solely on the existing classification systems to direct care. PMID- 14560900 TI - Nonsurgical treatment of adult acquired flat foot deformity. AB - The adult acquired flat foot requires assessment with unique casting and fabrication of support appliances. Conservative treatment is preferred as the initial protocol. Stabilization and control of affected joints can provide the patient with a decreased level of pain and an increased level of function. The orthosis design must acceptable to the patient's lifestyle to ensure compliancy. PMID- 14560901 TI - Nonoperative treatment of adult acquired flat foot with the Arizona brace. AB - Nonoperative treatment of posterior tibial tendon dysfunction can be successful with the Arizona AFO brace, particularly when treatment is initiated in the early stages of the disease. This mandates that the orthopedist has a high index of suspicion when evaluating patients to make an accurate diagnosis. Although there is a role for surgical management of acquired flat feet, a well-fitted, custom molded leather and polypropylene orthosis can be effective at relieving symptoms and either obviating or delaying any surgical intervention. In today's climate of patient satisfaction directed health care, a less invasive treatment modality that relieves pain may prove to be more valuable than similar pain relief that is obtained after surgery. Questions regarding the long-term results of bracing remain unanswered. Future studies are needed to determine if disease progression and arthrosis occur despite symptomatic relief with a brace. Furthermore, age- and disease stage-matched control groups who are randomized to undergo surgery or bracing are necessary to compare these different treatment modalities. At this time, the Arizona AFO brace can be a useful weapon in the orthopedist's armamentarium for treating acquired flat foot deformity. PMID- 14560902 TI - Soft tissue procedures. AB - Soft tissue procedures for PTT dysfunction and adult acquired flat foot improve function and preserve joint motion. These procedures can only be applied to patients who have correctable deformities. The durability of these procedures, without the addition of bone realignment procedures, has been questioned and needs to be investigated further. Reconstruction of the spring ligament complex corrects the flat foot in cadavers but has not been studied clinically. There is still much to be learned in this condition so we are able to provide optimal care for our patients. PMID- 14560903 TI - Acute posttraumatic planovalgus foot deformity involving hindfoot ligamentous pathology. AB - This article outlines the pathology of posttraumatic adult flat foot deformity that involves hindfoot ligamentous injury. The clinical presentation, radiographic findings, and pathology that are noted at the time of reconstruction are described. A proposed surgical reconstruction is presented that has successfully reconstructed the longitudinal arch of the affected foot. PMID- 14560905 TI - Lateral-sided bony procedures. AB - Treatment of any hindfoot deformity should include correction of the deformity and preservation of complex hindfoot motion. This important motion is protective of adjacent, and more removed, joints in that it serves a shock-absorbing function and protects them from stresses. Lateral column lengthening combined with a medial soft-tissue procedure is the treatment of choice for stage II flat foot. Patients who have significant subluxation of the subtalar joint will also need a medial displacement calcaneal osteotomy to correct the hindfoot valgus. Only patients who have a rigid foot secondary to degenerative changes will require an arthrodesis to correct the deformity and provide pain relief. Unfortunately, although fusion works well to correct deformity, it accelerates future degenerative changes. PMID- 14560904 TI - Medial-sided bony procedures: why, what, and how? AB - The adult acquired flat foot deformity is a common clinical entity; rupture or incompetence of the posterior tibial tendon is a frequent cause. The natural history is characterized by progressively worsening deformity and early recognition is important. Nonoperative treatment can alleviate symptoms and control progression in nearly all stages of the disease. Should this fail to control symptoms or prevent progression of deformity, operative intervention should be considered. In stage I disease, exploration and debridement, with or without FDL tendon transfer, is a viable option. In stage II disease, the PTT becomes elongated and the medial soft tissues become attenuated. Exploration and debridement of the PTT is performed, but frequently a FDL tendon transfer or side to-side anastomosis is required. It has been shown that soft tissue procedures alone may fail to correct deformity and this can lead to deterioration of results over time. Combined procedures, including soft tissue reconstructions to restore PTT function and bony procedures to correct deformity, have become popular. When the PTT is intact and degeneration or elongation is minimal, as in stage I or early stage II disease, reconstruction of the medial column with advancement of an osteoperiosteal flap based on the PTT insertion, combined with selective arthrodeses of the medial column, may be considered. These procedures have been well described for the treatment of symptomatic flexible flat foot in children and adolescents but experience in adults is lacking. Although it may be theoretically possible to passively correct hindfoot valgus with these procedures, it seems prudent to limit the indications to patients who have early disease accompanied by an isolated midfoot sag. In more advanced stage II disease, correction of deformity with a tendon transfer combined with a medial displacement calcaneal osteotomy or a lateral column lengthening is currently recommended. This allows for correction of deformity while sparing the hindfoot joints, which may be particularly important in young or active patients. Short term studies showed excellent results, but long-term results are lacking. In stage III disease, in which the deformity is fixed, arthrodesis is the procedure of choice. Isolated talonavicular arthrodesis has been shown to correct nearly all aspects of the deformity with long-lasting results. This procedure results in nearly complete lack of hindfoot motion and may predispose the patient to adjacent joint arthrosis. In a patient who has stage III disease with arthrosis confined to the talonavicular joint, isolated talonavicular arthrodesis may be considered. This clinical situation is rare, and, in most patients, a triple arthrodesis is probably preferred. If residual deformity is present after these procedures, it must be addressed. Residual medial column instability may be addressed by adding a selective arthrodesis of the naviculo-cuneiform or first metatarsocuneiform joint, whereas residual forefoot varus or supination may be addressed with selected midfoot fusions with or without a cuneiform osteotomy. PMID- 14560906 TI - Hindfoot arthrodesis for the adult acquired flat foot. AB - This article reviews hindfoot arthrodesis for the adult acquired flat foot. Discussion of triple arthrodesis, triple arthrodesis with the addition of a lateral column lengthening procedure, triple arthrodesis with the addition of a plantar flexion first metatarsoncuneiform arthrodesis, and an isolated subtalar arthrodesis are discussed. The indications, surgical techniques, outcome studies, and complications are addressed. PMID- 14560907 TI - Controversies in surgical reconstruction of acquired adult flat foot deformity. AB - There is a wide variety of approaches to the surgical treatment of acquired adult flat foot deformity. There are controversies in several areas, including disagreement over the cause of the condition, the structures that are involved, and what is the most appropriate surgical procedure or combination of procedures to successfully treat this condition. Trends in the orthopedic foot and ankle community include an attempt to preserve the hindfoot joints to allow for more normal biomechanics, the correction of bony deformities so that soft tissues are not overloaded, and an attempt to address the underlying cause so that the propensity for the deformity to recur is lessened. PMID- 14560908 TI - Arthroerisis of the subtalar joint. AB - Arthroerisis of the subtalar joint can be a valuable tool for the foot and ankle surgeon in treating flexible pes planus deformity in adults, as well as in children. Whether by itself, or in conjunction with other procedures, such as Achilles tendon lengthening (to allow dorsiflexion to at least neutral), midfoot or hindfoot osteotomies (to correct significant bony deformities), and posterior tibial tendon advancements (with associated symptomatic accessory navicular), arthroerisis can be helpful to correct deformity. When compared with procedures that involve osteotomy or arthrodesis, there is less morbidity to the patient who undergoes arthoerisis during surgery as well as postoperatively. There is no risk of nonunion, less immobilization is usually required, and arthroerisis is technically easy to perform. As with all surgical procedures, proper patient selection is extremely important. PMID- 14560909 TI - Stage IV posterior tibial tendon insufficiency: the tilted ankle. AB - Stage IV PTTD is the most challenging of the posterior tibial tendon deficiencies. The combination of a flattened longitudinal arch and a tilted ankle make successful management unpredictable. Conservative management universally fails and surgical options have been limited to pantalar and tibiotalocalcaneal arthrodesis. Alternatives to surgical management included herein are unproven, but provide a potential solution beyond that of arthrodesis. PMID- 14560910 TI - Issues relating to failure in the treatment of posterior tibial tendon dysfunction. AB - As our understanding of the underlying cause of flat foot progression improves so will our understanding of how best to catalog and treat the instabilities that are seen. Determining the failure of one type of treatment will be difficult until we can better define the varied pathology and give the treatment methods sufficient time to prove or disprove their premise. If an underlying systemic disease, where either unreliable motor function or unstable ligament support is present, joint arthrodeses are the preferred method of treatment. It is important not to fuse in situ, but rather effect the complete realignment of the foot through the fused joints. At a minimum, I believe that the subtalar fusion is the most appropriate method for gaining a stable correction. Any more stability will warrant a triple arthrodesis. Treatment for the diseased tendon should also be addressed appropriately. Tendon reconstruction alone provides no assurance of weight-bearing deformity correction or lasting function for most patients who have isolated type 2 dysfunction and should be avoided as a stand-alone procedure. It still serves a vital purpose in restoring function to the foot, and, combined with other procedures, aids in the preservation of dynamic response to weight-bearing loads. The choice of adjunctive procedures should be based on the pathology present. Each of the treatment regimens that is discussed in this article has a place in the overall treatment of the clinical disease but none, by itself, seems to correct all of the presentations that are possible for this pathologic entity. The possible exception may be the use of a subtalar realignment and fusion, but this may be overkill in many mild to moderate deformities. Care should be taken in choosing the proper treatment based on the pathology that is presented by each patient. PMID- 14560911 TI - The treatment of failed reconstruction for adult acquired flat foot deformity. AB - The successful salvage of a failed reconstruction for adult acquired flat foot deformity requires a thorough assessment of the alignment, healing, and function of the entire foot. The pain that is experienced by the patient often emanates from multiple sources. Once identified, the plan of treatment must provide an integrated and comprehensive approach to resolve the origins of pain to restore durable function to the foot. Often, these goals may be achieved through the use of orthotics, physical therapy, and other nonoperative means. Additional or revision surgical procedures may be necessary to complete or correct the previous attempts to reconstruct the foot. PMID- 14560912 TI - Mechanism studies on the CSI reaction with allyl ethers by varying p-substituent. AB - We examined the effect of p-substituents in p-substituted cinnamyl methyl ethers and 1-(p-substituted phenyl)allyl methyl ethers with CSI, and confirmed that the CSI reaction of allyl ethers (p-substituted ethers) is a competitive reaction of S(N)i and S(N)1 mechanism according to the stability of the carbocation. And, the only terminal allylic amine was obtained through the migration reaction in thermodynamic reaction condition. PMID- 14560913 TI - Synthesis and antiviral activity of novel C-methyl branched cyclopropyl nucleosides. AB - A series of novel cyclopropyl nucleosides was synthesized using the highly stereoselective Simmons-Smith reaction starting from 1,2:5,6-di-O-isopropylidene D-mannitol. The structural assignments of these nucleosides were determined by NMR studies and X-ray crystallography. All the synthesized nucleosides were assayed against several viruses. PMID- 14560914 TI - Synthesis and biological investigations of new thiazolidinone and oxadiazoline coumarin derivatives. AB - Ethyl (coumarin-4-oxy)acetate 1 was prepared through the reaction of 4 hydroxycoumarin with ethyl bromoacetate. Compound 1 was allowed to react with hydrazine hydrate to produce coumarin-4-oxyacetic hydrazide 2. The synthesis of N (arylidene and alkylidene)-coumarin-4-oxyacetic hydrazones 3-20 was performed. The preparation of 2-substituted-3-[(coumarin-4-oxy) acetamido]thiazolidinones 21 26 and 2-[(coumarin-4-oxy)methyl]-4-acetyl-5-substituted-delta2-1,3,4 oxadiazolines 27-33 was performed by the reaction of the hydrazones 3, 4, 7, 9, 12, 14 with mercaptoacetic acid and the hydrazones 3, 4, 5, 7, 12, 15, 16 with acetic anhydride, respectively. The antiviral activities, cytotoxicities and structure-activity relationship (SAR) towards different microorganisms of the prepared compounds were studied. PMID- 14560915 TI - Combined isobutoxycarbonylation and tert-butyldimethylsilylation for the GC/MS SIM detection of alkylphenols, chlorophenols and bisphenol A in mackerel samples. AB - The alkylphenols, chlorophenols, and bisphenol A were determined by gas chromatography/mass spectrometry-selected ion monitoring (GC/MS-SIM) followed by two work-up methods for comparison: isobutoxycarbonyl (isoBOC) derivatization and tert-butyldimethylsilyl (TBDMS) derivatization. Eleven endocrine disrupting chemicals (EDCs) of phenols in biological samples were extracted with acetonitrile and then the acetonitrile layer underwent freezing filtration 60 degrees C for 2 hours. Solid-phase extraction (SPE) was used with XAD-4 and subsequent conversion to isoBOC or TBDMS derivatives for sensitivity analysis with the GC/MS-SIM mode. For isoBOC derivatization and TBDMS derivatization the recoveries were 92.3 approximately 150.6% and 93.8-108.3%, the method detection limits (MDLs) of bisphenol A for SIM were 0.062 microg/kg and 0.010 microg/kg, and the SIM responses were linear with the correlation coefficient varying by 0.9755-0.9981 and 0.9908-0.9996, respectively. When these methods were applied to mackerel samples, the concentrations of the 11 phenol EDCs were below the MDL. PMID- 14560916 TI - A new 24-nor-lupane-glycoside of Acanthopanax trifoliatus. AB - A new 24-nor-lupaneglycoside was isolated from the leaves of Acanthopanax trifoliatus. Based on spectroscopic data its chemical structure was determined as 24-nor-11alpha-hydroxy-3-oxo-lup-20(29)-en-28-oic acid 28-O-alpha-L rhamnopyranosyl-(1 --> 4)-beta-D-glucopyranosyl-(1 --> 6)-beta-D-glucopyranosyl ester. PMID- 14560917 TI - Alkaloidal constituents from Aconitum jaluense. AB - Aconitum jaluense Komar. (Ranunculaceae) is one of the Aconitum plants growing in Korean peninsula. An investigation of the alkaloidal constituents of this species led to the isolation of seven C19-norditerpenoid and a C20-diterpenoid alkaloid. Three of them have been identified as neoline, mesaconitine, and hypaconitine, which were isolated from this plant collected from Mt. Bultasan in the north part. The other five alkaloids were determined as lipomesaconitine, lipohypaconitine, 15alpha-hydroxyneoline, hokbusine A, and napelline, which have not been found in this plant. Structures of those alkaloids were determined on the basis of their spectral data. It is of interest to note that a comparison of the present work and the previous report showed some differences in the alkaloidal contents. PMID- 14560918 TI - Pheophorbide A-methyl ester, Acyl-CoA: cholesterol acyltransferase inhibitor from Diospyros kaki. AB - In the course of our search for Acyl-CoA: cholesterol acyltransferase (ACAT) inhibitors from natural sources, a new type of ACAT inhibitor was isolated from a methanol extract of Diospyros kaki. On the basis of spectral and structural evidence, the compound was identified as pheophorbide A-methyl ester. Pheophorbide A-methyl ester inhibited ACAT activity in a dose dependent manner with an IC50 value of 1.85 microg/mL. PMID- 14560919 TI - Anti-oxidant activities of fucosterol from the marine algae Pelvetia siliquosa. AB - The anti-oxidant activities of fucosterol isolated from the marine algae Pelvetia siliquosa were investigated. Fucosterol exhibited a significant decrease in serum transaminase activities elevated by hepatic damage induced by CCl4-intoxication in rats. Fucosterol inhibited the sGOT and sGPT activities by 25.57 and 63.16%, respectively. Fucosterol showed the increase in the anti-oxidant enzymes such as hepatic cytosolic superoxide dismutase (SOD), catalase and glutathione peroxidase (GSH-px) activities by 33.89, 21.56 and 39.24%, respectively, in CCl4-intoxicated rats. These results suggest that fucosterol possess not only the anti-oxidant, but also the hepatoprotective activities in rats. PMID- 14560920 TI - Platelet anti-aggregatory effects of coumarins from the roots of Angelica genuflexa and A. gigas. AB - Five coumarins, isoimperatorin (1), pabulenol (2), isooxypeucedanin (3), oxypeucedanin hydrate (4) and osthol (5) were isolated from the MeOH extract of Angelica genuflexa in the course of searching for anti-platelet and anti coagulant components from plants. Pabulenol (2) was isolated from A. genuflexa for the first time. The five compounds isolated from A. genuflexa, together with decursinol angelate (6), decursin (7) and nodakenin (8) from A. gigas were evaluated for their effects on platelet aggregation and blood coagulation. Compounds 2, 5, 6 and 7 were observed to be either equally effective or 2-4 times more inhibitory than ASA in both arachidonic acid and U46619 (TXA2 mimetic) induced platelet aggregations. PMID- 14560921 TI - Anti-tumor activities of decursinol angelate and decursin from Angelica gigas. AB - The in vivo anti-tumor activities of decursinol angelate (1) and decursin (2) isolated from the roots of Angelica gigas were investigated. These two compounds, when administered consecutively for 9 days at 50 and 100 mg/kg i.p. in mice, caused a significant increase in the life span and a significant decrease in the tumor weight and volume of mice inoculated with Sarcoma-180 tumor cells. These results suggest that decursinol angelate (1) and decursin (2) from A. gigas have anti-tumor activities. PMID- 14560922 TI - Inhibitory effect of kaurane type diterpenoids from Acanthopanax koreanum on TNF alpha secretion from trypsin-stimulated HMC-1 cells. AB - Five known kaurane type diterpenoids, 16alphaH,17-isovaleryloxy-ent-kauran-19-oic acid (1), 16alpha-hydroxy-17-isovaleryloxy-ent-kauran-19-oic acid (2), paniculoside-IV (3), 16alpha-hydroxy-ent-kauran-19-oic acid (4), and ent-kaur-16 en-19-oic acid (5) were isolated from the root of Acanthopanax koreanum by repeated column chromatography and reversed phase preparative HPLC. The structures of these compounds were established from physicochemical and spectral data. Among the isolated compounds 16alphaH,17-isovaleryloxy-ent-kauran-19-oic acid (1) showed potent inhibitory activity (IC50 value, 16.2 uM) on TNF-alpha secretion from HMC-1, a trypsin-stimulated human leukemic mast cell line. PMID- 14560923 TI - Inhibitory effect of trans-N-p-coumaroyl tryamine from the twigs of Celtis chinensis on the acetylcholinesterase. AB - The methanolic extract of the twigs of Celtis chinensis was found to show inhibitory activity on acetylcholinesterase (AChE), an enzyme that plays a role in the metabolic hydrolysis of ACh. Bioassay-guided fractionation of the methanolic extract resulted in the isolation of N-p-coumaroyl tyramine, as an inhibitor on AChE. This compound inhibited AChE activity in a dose-dependent manner, and the IC50 value of trans-N-p-coumaroyl tyramine was 34.5 microg/mL (122 microM). PMID- 14560924 TI - Sphingosine-1-phosphate-induced ERK activation protects human melanocytes from UVB-induced apoptosis. AB - Ultraviolet B (UVB) is known to induce apoptosis in human melanocytes. Here we show the cytoprotective effect of sphingosine-1-phosphate (S1P) against UVB induced apoptosis. We also show that UVB-induced apoptosis of melanocytes is mediated by caspase-3 activation and poly(ADP-ribose) polymerase (PARP) cleavage, and that S1P prevents apoptosis by inhibiting this apoptotic pathway. We further investigated three major mitogen-activated protein (MAP) kinases after UVB irradiation. UVB gradually activated c-Jun N-terminal kinase (JNK) and p38 MAP kinase, while extracellular signal-regulated protein kinase (ERK) was inactivated transiently. Blocking of the p38 MAP kinase pathway using SB203580 promoted cell survival and inhibited the activation of caspase-3 and PARP cleavage. These results suggest that p38 MAP kinase activation may play an important role in the UVB-induced apoptosis of human melanocytes. To explain this cytoprotective effect, we next examined whether S1P could inhibit UVB-induced JNK and p38 MAP kinase activation. However, S1P was not found to have any influence on UVB induced JNK or p38 MAP kinase activation. In contrast, S1P clearly stimulated the phosphorylation of ERK, and the specific inhibition of the ERK pathway using PD98059 abolished the cytoprotective effect of S1P. Based on these results, we conclude that the activation of p38 MAP kinase plays an important role in UVB induced apoptosis, and that S1P may show its cytoprotective effect through ERK activation in human melanocytes. PMID- 14560925 TI - Cotinine inhibits catecholamine release evoked by cholinergic stimulation from the rat adrenal medulla. AB - The aim of the present study was to clarify whether cotinine affects the release of catecholamines (CA) from the isolated perfused rat adrenal gland, and to establish the mechanism of its action, in comparison with the response of nicotine. Cotinine (0.3-3 mM), when perfused into an adrenal vein for 60 min, inhibited CA secretory responses evoked by ACh (5.32 mM), DMPP (a selective neuronal nicotinic agonist, 100 microM for 2 min) and McN-A-343 (a selective muscarinic M1-agonist, 100 microM for 2 min) in dose- and time-dependent manners. However, cotinine did not affect CA secretion by high K+ (56 mM). Cotinine itself also failed to affect basal CA output. Furthermore, in the presence of cotinine (1 mM), CA secretory responses evoked by Bay-K-8644 (an activator of L-type Ca2+ channels, 10 microM) and cyclopiazonic acid (an inhibitor of cytoplasmic Ca2+ ATPase, 10 microM) were relative time-dependently attenuated. However, nicotine (30 microM), given into the adrenal gland for 60 min, initially rather enhanced CA secretory responses evoked by ACh and high K+, followed by the inhibition later, while it time-dependently depressed the CA release evoked by McN-A-343 and DMPP. Taken together, these results suggest that cotinine inhibits greatly CA secretion evoked by stimulation of cholinergic (both nicotinic and muscarinic) receptors, but does fail to affect that by the direct membrane-depolarization. It seems that this inhibitory effect of cotinine may be exerted by the cholinergic blockade, which is associated with blocking both the calcium influx into the rat adrenal medullary chromaffin cells and Ca2+ release from the cytoplasmic calcium store. It also seems that there is a big difference in the mode of action between cotinine and nicotine in the rat adrenomedullary CA secretion. PMID- 14560926 TI - Assessment of the estrogenicity of isoflavonoids, using MCF-7-ERE-Luc cells. AB - In the current study, our research focused on the estrogenic activity of isoflavonoids, mainly genistein, biochanin A and daidzein. Genistein enhanced the reporter gene expression of MCF-7-ERE-Luc cells, at a concentration as low as 10 nM, with a concentration of 100 nM the achieved gene expression effects were similar to those of 10 pM 17beta-estradiol, Based on the estrogenic activities of biochanin A and daidzein, hydroxyl groups at the 4' and 5 positions are needed for the maximal effect of the genistein. The estrogenic effects of these isoflavonoids were inhibited by the concomitant treatment with tamoxifen. The data showed that the estrogenic effects of isoflavonoids were mediated through estrogen receptors. When the isoflavonoids were tested as mixtures, the estrogenic effects were lower than the arithmetic sum of those induced by each individual isoflavonoid. The estrogenic potency of each isoflavonoid was presented at EC50 levels with a 17beta-estradiol equivalent concentration (EEQ) based on the dose response of each chemical. The EC50s and EEQs of genistein, biochanin A and daidzein were 4.15, 0.89 and 0.18 microM, and 15.0, 5.12 and 1.83 microM/M, respectively. Our data clearly demonstrated that the pERE-luciferase reporter gene assay was suited for the sensitive and quantitative measurement, and large scale screening, of the estrogenicity of chemicals in vitro. PMID- 14560927 TI - Pharmacokinetic behavior and tissue distribution of verapamil and its enantiomers in rats by HPLC. AB - The differences in pharmacokinetic behavior and tissue distribution of verapamil and its enantiomers were investigated in rats. In high-performance liquid chromatographic method, an achiral ODS column (150 mm x 4.6 mm i.d.) with the mobile phase consisting of methanol-water (73:30, v/v) was used for the determination of the concentration for racemic verapamil, and a Chiralcel OJ column (250 mmx4.6 mm i.d.) with the mixture of n-haxane-ethanol-triethylamine (85:15:0.2, v/v/v) as mobile phase was used to determine the concentrations of verapamil enantiomers. A fluorescence detector in the analytical system was set at excitation and emission wavelengths of 275 nm and 315 nm. The differences between enantiomers were apparent in the pharmacokinetics in rats. The area under the concentration-time curve (AUC) of S-(-) verapamil was higher than that of R (+) verapamil. The half-distribution time (T 1/2(alpha)) of S-(-) verapamil which distributing to tissue from blood was shorter than that of R-(+) verapamil, but the elimination half-time (T 1/2(beta)) was longer in rat following oral administration of racemic verapamil. At 1.3 h after oral administration of racemic verapamil, however, there were no significant differences between enantiomers for the distributions in major tissues such as heart, cerebrum, cerebellum, liver, spleen and kidney. PMID- 14560928 TI - Commonly used surfactant, Tween 80, improves absorption of P-glycoprotein substrate, digoxin, in rats. AB - Tween 80 (Polysorbate 80) is a hydrophilic nonionic surfactant commonly used as an ingredient in dosing vehicles for pre-clinical in vivo studies (e.g., pharmacokinetic studies, etc.). Tween 80 increased apical to basolateral permeability of digoxin in Caco-2 cells suggesting that Tween 80 is an in vitro inhibitor of P-gp. The overall objective of the present study was to investigate whether an inhibition of P-gp by Tween 80 can potentially influence in vivo absorption of P-gp substrates by evaluating the effect of Tween 80 on the disposition of digoxin (a model P-gp substrate with minimum metabolism) after oral administration in rats. Rats were dosed orally with digoxin (0.2 mg/kg) formulated in ethanol (40%, v/v) and saline mixture with and without Tween 80 (1 or 10%, v/v). Digoxin oral AUC increased 30 and 61% when dosed in 1% and 10% Tween 80, respectively, compared to control (P < 0.05). To further examine whether the increase in digoxin AUC after oral administration of Tween 80 is due, in part, to a systemic inhibition of digoxin excretion in addition to an inhibition of P-gp in the GI tract, a separate group of rats received digoxin intravenously (0.2 mg/kg) and Tween 80 (10% v/v) orally. No significant changes in digoxin IV AUC was noted when Tween 80 was administered orally. In conclusion, Tween 80 significantly increased digoxin AUC and Cmax after oral administration, and the increased AUC is likely to be due to an inhibition of P-gp in the gut (i.e., improved absorption). Therefore, Tween 80 is likely to improve systemic exposure of P-gp substrates after oral administration. Comparing AUC after oral administration with and without Tween 80 may be a viable strategy in evaluating whether oral absorption of P-gp substrates is potentially limited by P-gp in the gut. PMID- 14560929 TI - Aspirin resistance: myth or reality? PMID- 14560930 TI - Stress echocardiography for the diagnosis of coronary artery disease. PMID- 14560931 TI - Tuberculous pericardial effusion--mediastinal lymph glands: the cause and clue to the etiology. AB - BACKGROUND: Tuberculous pericardial effusion is most often due to the spread of tuberculosis from the mediastinal lymph glands; however, no attempt has yet been made to study these glands. We studied the mediastinal glands in proven tuberculous pericardial effusion patients and hypothesized that the findings may be of use in the etiological diagnosis of pericardial effusion. METHODS AND RESULTS: We studied 45 patients with large pericardial effusion or tamponade. All underwent chest computed tomographic studies that were reviewed by radiologists blinded to the diagnosis. Of these 45 patients, 27 had tuberculosis and 18 had viral or idiopathic effusion. Pericardial biopsy was done in 25/27 and tuberculin skin test in 22/27 patients with tuberculosis, and all received specific treatment. In patients with tuberculosis the skin test measured 17+/-3.3 mm. All 27 had mediastinal lymph glands > or = 10 mm in size. The mean size of the mediastinal glands was 19.5+/-8.6 mm and the mean number was 2.5+/-1.2. The aortopulmonary glands were the most frequently enlarged (63%), and hilar the least often (14.8%). The glands showed a hypodense center in 52% of the patients. On follow-up of 15.8+/-10.4 months, glands were not seen in 80.9%, and were smaller in size in 19%; none had a hypodense center. Marked lymphadenopathy was not seen in any patient with viral/idiopathic pericardial effusion. Two had glands < or = 5 mm in size. CONCLUSIONS: Only patients with tuberculosis had substantial mediastinal lymph gland enlargement and not those with viral or idiopathic pericardial effusion. Such glands disappeared or regressed on treatment. In the appropriate clinical context, marked nonhilar mediastinal lymphadenopathy on chest computed tomographic studies along with a strongly positive tuberculin skin test could be of value in the noninvasive diagnosis of pericardial effusion due to tuberculosis. PMID- 14560932 TI - A tertiary care hospital-based study of conventional risk factors including lipid profile in proven coronary artery disease. AB - BACKGROUND: The prevalence and mortality rates of coronary artery disease have been known to be higher in the Indian than the Western population. Most data on lipid levels in Indians have been obtained from studies on migrant Asian Indians. There are insufficient data on lipid profile and other conventional risk factors in Indian patients living within India. METHODS AND RESULTS: The study included 2656 consecutive patients who underwent coronary angiography between March 1998 and February 2002. Of these, 2399 subjects had angiographically proven coronary artery disease (group 1) while 257 had normal coronary arteries (group 2). Lipid values were measured in the fasting state on the morning the coronary angiography was done. Patients receiving lipid-lowering agents, those having renal, hepatic or thyroid disorders, patients presenting within 8 weeks after acute myocardial infarction, and patients who were taking noncardiac drugs that affect the lipid profile were excluded from the study. Other conventional risk factors were also recorded. In subjects with coronary artery disease and normal coronary arteries, the levels of mean total cholesterol recorded were 178.5+/-42.1 mg/dl v. 154.1+/ 40.2 mg/dl (p<0.001), high-density lipoprotein cholesterol 30.6+/-9 mg/dl v. 27.3+/-6.8 mg/dl (p<0.001), low-density lipoprotein cholesterol 109.8+/-35.4 mg/dl v 93.6+/-33.9 mg/dl (p<0.001), and triglyceride 190.7+/-95.4 mg/dl v. 157.6+/-73.5 mg/dl (p<0.001), respectively. In subgroup analysis by age, the younger coronary artery disease group (< or = 40 years) had significantly higher total and low-density lipoprotein cholesterol levels than the older group (> 40 years), viz. 194.6+/-51.4 mg/dl v. 176.3+/-40.2 mg/dl (p<0.001), and 118.3+/-39.6 mg/dl v. 108.7+/-36.1 mg/dl (p=0.001). Triglyceride levels were not significantly different [211.7+/-105.1 mg/dl v. 187.8+/-93.6 mg/dl (p=ns)], being equally high in both subgroups and, although high-density lipoprotein cholesterol levels were different, this difference was minimal, being equally low in both [32.7+/-9.5 mg/dl v. 30.3+/-9.0 mg/dl (p=ns)]. In the subgroup analysis of those with coronary artery disease, diabetics had significantly lower total cholesterol [174+/-41.1 mg/dl v. 180.4+/-42.4 mg/dl (p<0.001)] and low-density lipoprotein cholesterol levels [105.8+/-34 mg/dl v. 111.5+/-35.8 mg/dl (p<0.001)] than non diabetics, whereas triglyceride and high-density lipoprotein cholesterol levels were not significantly different, triglycerides being equally high in both [186.2+/-95.5 mg/dl v. 192.5+/-95.2 mg/dl (p=ns)], and high-density lipoprotein equally low in both [30.9+/-9.3 mg/dl v. 30.5+/-9 mg/dl (p=ns)]. The commonest associated conventional risk factor in diabetics was hypertension and, in the younger age group (< or = 40 years), it was smoking and a positive family history of premature coronary artery disease. CONCLUSIONS: We conclude that in north Indians, coronary artery disease occurs at much lower levels of total cholesterol and low-density lipoprotein cholesterol than other populations, and high triglyceride and low high-density lipoprotein levels are more of a universal phenomenon in this population. Younger patients have a more atherogenic lipid profile than the older subgroup with coronary artery disease, and smoking and a family history of premature coronary artery disease are the most common associated risk factors. Total cholesterol levels seem to play a lesser role in the occurrence of coronary artery disease in diabetics, the presence of which is in itself overwhelming for the occurrence of coronary artery disease. PMID- 14560933 TI - Carotid artery stenting with filter protection. AB - BACKGROUND: Neurologic events associated with distal embolization of debris during percutaneous carotid artery stenting complicate the procedure. Filter devices for cerebral protection potentially reduce the risk of embolization and other neurologic events. We studied the feasibility, safety, and efficacy of carotid artery stenting with a filter device. METHODS AND RESULTS: Between January 2002 and January 2003, a total of 22 consecutive patients (30 lesions) who had >70% diameter stenosis of the internal carotid artery underwent carotid artery stenting with filter protection at our institute. The mean age of the patients was 64+/-9 years; 14 were men and 8 women, and 15 had neurologic symptoms. A stent was successfully implanted in 29 lesions. It was possible to position a filter device in all the 29 lesions. Neurologic complications during the procedure, in the hospital, and at 30-day clinical follow-up occurred in 2 patients. One patient suffered a minor stroke that resolved within 24 hours. None of the patients had a major embolic stroke. There was one death from intracerebral hemorrhage related to hyperperfusion and the use of a glycoprotein IIb/IIIa inhibitor. CONCLUSIONS: Filter protection during carotid artery stenting seems technically feasible, safe, and effective. In the present study, the incidence of embolic neurologic events was low. PMID- 14560934 TI - Prevalence, correlates, awareness, treatment, and control of hypertension in a middle-aged urban population in Kerala. AB - BACKGROUND: Hypertension is an important cause of cardiovascular morbidity and mortality. METHODS AND RESULTS: We conducted a cross-sectional survey of 314 middle-aged subjects (163 men; age range 40-60 years, mean 49 years) in urban Thiruvananthapuram City. Kerala, to estimate the prevalence of hypertension, examine its correlates, and assess the degree of awareness, treatment, and control of high blood pressure. Blood pressure was measured by a nurse graduate using a mercury column sphygmomanometer and a standardized technique. We used multivariable analyses to examine the sociodemographic and clinical correlates of hypertension. The overall prevalence of hypertension in our sample was 54.5% (men 56.3%, women 52.3%). The factors associated with an increased prevalence of hypertension were higher body-mass index (odds ratio for a value in the top tertile of 2.33, 95% confidence interval: 1.2-4.4), and older age (odds ratio for the age group 55-60 years of 2.65, 95% confidence interval: 1.3-5.6). An occupation involving moderate or greater physical activity was inversely associated with the prevalence of hypertension (odds ratio 0.35, 95% confidence interval 0.13-0.94). Among hypertensives, 39% were aware of the condition, while 29% were treated with blood pressure-lowering medications. Adequate control of elevated blood pressure was achieved in only 30.6% of treated hypertensives. In our community-based sample, over half of all middle-aged individuals were hypertensive, but less than a third were under treatment. Adequate control of hypertension was achieved in less than a third of the treated individuals. CONCLUSIONS: These observations re-emphasize the need for hypertension awareness programs targeting the general public and the increased use of opportunistic blood pressure screening, and underscore the importance of measures to increase the knowledge of current guidelines for the detection and treatment of hypertension among healthcare providers. PMID- 14560936 TI - Transcatheter closure of perimembranous ventricular septal defect in a patient with situs inversus and dextrocardia. AB - Successful transcatheter closure of a perimembranous ventricular septal defect with an Amplatzer device has been reported in patients with levocardia. We report a case in which the device could be deployed successfully in a child with isolated perimembranous ventricular septal defect with situs inversus and dextrocardia. PMID- 14560935 TI - Functional proteomics of receptor-Ck in the developmental stages of human atherosclerotic arterial wall. AB - BACKGROUND: The study was undertaken to understand the relationship between the functional proteomics of receptor-Ck and developmental stages of human atherosclerotic aortic wall. METHODS AND RESULTS: Gene expression study of 25 aortas was undertaken and the results revealed a gradual increase in receptor-Ck gene expression paralleled by the regulatory response of its effector genes coding for sterol response element-binding protein, p27, cyclin D, interleukin-6 and CD40 from a normal to atherosclerotic arterial wall (viz. fatty streak and fibrofatty/fibrous plaque). CONCLUSIONS: Based upon this and our earlier studies, we propose that cholesterol-specific receptor-Ck-dependent gene regulation may be of crucial importance in atherogenesis. PMID- 14560937 TI - Refractory adenosine-sensitive congenital His bundle tachycardia: response to calcium-channel blockers. AB - The congenital form of His bundle tachycardia is an uncommon pediatric arrhythmia. We report the case of a 7-year-old child with tachycardiomyopathy. The incessant arrhythmia, detected in infancy, was resistant to amiodarone and beta-blockers. During electrophysiologic study, the tachycardia converted to sinus rhythm with intravenous adenosine and diltiazem. Subsequently, the child is maintaining sinus rhythm on oral verapamil. Calcium-channel blockers should be considered for the treatment of this arrhythmia, which is often resistant to multiple antiarrhythmic drugs. PMID- 14560938 TI - Stenting of a large thrombus-containing subclavian artery stenosis using a distal protection device. AB - Atheromatous obstructive lesions of the arch vessels that contain thrombi are at high risk for distal embolization during angioplasty. This can lead to catastrophic neurological complications. We report a case of acute-on-chronic ischemia of the left upper limb due to thrombus-containing subclavian artery stenosis. After placement of an intravascular filter device, angioplasty and stent implantation successfully relieved the stenosis without any complications. PMID- 14560939 TI - Sutureless patch repair of post-myocardial infarction left ventricular rupture. AB - Ventricular rupture following myocardial infarction is a serious clinical problem with a high mortality. A 60-year-old man with left ventricular rupture and cardiac tamponade following myocardial infarction was managed successfully by emergency surgery. An onlay patch of Teflon held in place by an adhesive without any sutures was used to repair the ruptured myocardium. PMID- 14560940 TI - Acquired aortopulmonary fistula. AB - An acquired aortopulmonary artery fistula is rare. We describe a case with an aortic arch aneurysm communicating with the main pulmonary artery. The diagnosis was made on the basis of transthoracic echocardiography and confirmed by transesophageal echocardiography. A post-mortem examination revealed the complete anatomy of the aneurysm and the aortopulmonary communication. PMID- 14560941 TI - Exclusive carbon dioxide-guided renal artery stenting in a case of Takayasu's arteritis with a solitary functioning kidney. AB - We report the case of a 28-year-old female with type II Takayasu's arteritis affecting her single functioning kidney. Impaired renal function precluded the use of conventional contrast media. We used carbon dioxide-guided renal angioplasty to successfully treat the patient. PMID- 14560942 TI - Off-pump coronary artery bypass grafting in high-risk patients. PMID- 14560943 TI - Cardiovascular images. Double-orifice mitral valve. PMID- 14560944 TI - QRS widening as a marker of dys-synchrony. PMID- 14560945 TI - Brachial artery flow-mediated dilatation for detecting subclinical atherosclerosis. PMID- 14560946 TI - Brachial artery flow-mediated dilatation in prediction of coronary artery disease in Indian subjects. PMID- 14560947 TI - Sildenafil in pulmonary hypertension. PMID- 14560948 TI - TOR: the first 10 years. AB - TOR was discovered and christened 10 years ago. On the occasion of this anniversary, we revisit the discovery of TOR and chronicle subsequent breakthroughs in S. cerevisiae that contributed to an understanding of TOR function in yeast and higher eukaryotes. In particular, we discuss findings that led to the realization that the function of TOR is to control cell growth in response to nutrients. PMID- 14560949 TI - The role of phosphatases in TOR signaling in yeast. AB - The TOR pathway controls cellular functions necessary for cell growth and proliferation of yeast and larger eukaryotes. The search for members of the TOR signaling cascade in yeast led to the discovery of type 2A protein phosphatases (PP2A) as important players within the pathway. We describe the roles in yeast of PP2A and the closely related phosphatase, Sit4, and then focus on complexes formed between the catalytic subunit of these phosphatases and Tap42, a direct target of the Tor protein kinases in yeast. Recent results suggest that Tap42 mediates many of the Tor functions in yeast, especially those involved in transcriptional modulation. However, whether Tap42 executes its function by inhibiting phosphatase activity or by activating phosphatases is still uncertain. In addition, Tor affects some transcriptional and physiological processes through Tap42 independent pathways. Thus, Tor proteins use multiple mechanisms to regulate transcriptional and physiological processes in yeast. PMID- 14560950 TI - Yeast TOR signaling: a mechanism for metabolic regulation. AB - Understanding how cell growth is regulated in response to environmental signals remains a challenging biological problem. Recent studies indicate the TOR (target of rapamycin) kinase acts within an intracellular regulatory network used by eukaryotic cells to regulate their growth according to nutrient availability. This network affects all aspects of gene expression, including transcription, translation, and protein stability, making TOR an excellent candidate as a global regulator of cellular activity. Here we review our recent studies of two specific transcriptional outputs controlled by TOR in the budding yeast, S. cerevisiae: (1) positive regulation of genes involved in ribosome biogenesis, and (2) negative regulation of genes required for de novo biosynthesis of glutamate and glutamine. These studies have raised the important issue as to how diverse nutritional cues can pass through a common signaling pathway and yet ultimately generate distinct transcriptional responses. PMID- 14560951 TI - Nutrient signaling through TOR kinases controls gene expression and cellular differentiation in fungi. AB - The TOR kinases were first identified in Saccharomyces cerevisiae as the targets of the immunosuppressive drug rapamycin. Subsequent studies employing rapamycin as a tool in yeast have given us insight into the structure and function of the TOR kinases, as well as the biological role of the TOR signaling program in transmitting nutrient signals to promote cell growth. One of the major advances from this area has been in defining an unexpected role for TOR signaling in the regulation of transcription. The identification of target genes subject to regulation by TOR has provided a platform for the dissection of the signaling events downstream of the TOR kinases. Studies aimed at understanding TOR regulated transcription have begun to shed light on how TOR signaling cooperates with other signaling programs. In addition, the TOR pathway regulates the developmental program of pseudohyphal differentiation in concert with highly conserved MAP kinase and PKA signaling programs. Remarkably, rapamycin also blocks filamentation in a number of important human and plant pathogens and the mechanism of rapamycin action is conserved in Candida albicans and Cryptococcus neoformans. The antimicrobial properties of less immunosuppressive analogs of rapamycin hold promise for the development of an effective antifungal therapy. PMID- 14560952 TI - Autophagy in yeast: a TOR-mediated response to nutrient starvation. AB - TOR plays a key role in cell growth and cell-cycle progression, but in addition recent studies have shown that TOR is also involved in the regulation of a number of molecular processes associated with nutrient deprivation, such as autophagy. In budding yeast, TOR negatively regulates activation of Apg1 protein kinase, which is essential for the induction of autophagy. This review describes recent research in this field and the mechanism by which TOR mediates induction of autophagy. PMID- 14560953 TI - The fission yeast TOR proteins and the rapamycin response: an unexpected tale. AB - The TOR proteins are known as key regulators of cell growth in response to nutritional and mitogenic signals and as targets for the immunosuppressive and anti-cancerous drug rapamycin. The fission yeast Schizosaccharomyces pombe has two TOR homologues, tor1+ and tor2+. Despite their structural similarity, these genes have distinct functions: tor1+ is required under starvation, extreme temperatures, and osmotic or oxidative stress conditions, whereas tor2+ is required under normal growth conditions. Surprisingly, rapamycin does not seem to inhibit the S. pombe TOR-related functions. Rapamycin specifically inhibits sexual development in S. pombe, and this seems to stem from direct inhibition of the S. pombe FKBP12 homologue. Why S. pombe cells are resistant to rapamycin during the growth phase is as yet unclear and awaits further analysis of the TOR dependent signaling pathways. PMID- 14560954 TI - Plant growth and the TOR pathway. AB - In mammalian, insect, and yeast cells, TOR proteins are essential regulators of cell growth in response to environmental signals including nutrients, mitogens, and stresses. Although many aspects of the TOR-dependent signalling pathway are conserved between animals and fungi, important differences have also been found and are likely to be related to the ecophysiological adaptations of these organisms. The TOR protein also exists in plants. This review will first discuss specific aspects of plants concerning the contribution of cell growth to overall growth, as well as their responses to nutrient starvation, with emphasis on recent results obtained through genetic analysis in the model plant Arabidopsis thaliana. This is followed by the current status of the genetic analysis of the TOR gene in this plant and the search for potential members of a TOR pathway in the Arabidopsis genome. PMID- 14560955 TI - TOR action in mammalian cells and in Caenorhabditis elegans. AB - The p70 S6 kinase (p70 S6K) was the first signaling element in mammalian cells shown to be inhibited by rapamycin. The activity of the p70 S6K in mammalian cell is upregulated by extracellular amino acids (especially leucine) and by signals from receptor tyrosine kinases (RTKs), primarily through activation of the type 1A PI-3 kinase. The amino acid-/rapamycin-sensitive input and the PI-3 kinase input are co-dominant but largely independent, in that deletion of the amino terminal and carboxy-terminal noncatalytic sequences flanking the p70 S6K catalytic domain renders the kinase insensitive to inhibition by both rapamycin and by withdrawal of amino acids, whereas this p70 S6K mutant remains responsive to activation by RTKs and to inhibition by wortmannin. At a molecular level, this dual control of p70 S6K activity is attributable to phosphorylation of the two p70 S6K sites: The Ptd Ins 3,4,5P3-dependent kinasel (PDK1) phosphorylates p70 S6K at a Thr on the activation loop, whereas mTOR phosphorylates a Thr located in a hydrophobic motif carboxyterminal to the catalytic domain. Together these two phosphorylations engender a strong, positively cooperative activation of p70 S6K, so that each is indispensable for physiologic regulation. Like RTKs, the p70 S6K appears early in metazoan evolution and comes to represent an important site at which the more ancient, nutrient-responsive TOR pathway converges with the RTK/PI 3 kinase pathway in the control of cell growth. Dual regulation of p70 S6K is seen in Drosophila; however, this convergence is not yet evident in Caenorhabditis elegans, wherein nutrient activation of the insulin receptor (InsR) pathway negatively regulates dauer development and longevity, whereas the TOR pathway regulates overall mRNA translation through effectors distinct from p70 S6K, as in yeast. The C. elegans TOR and InsR pathways show none of the cross or convergent regulation seen in mammalian cells. The nature of the elements that couple nutrient sufficiency to TOR activity remain to be discovered, and the mechanisms by which RTKs influence TOR activity in mammalian cells require further study. One pathway for RTK control involves the tuberous sclerosis complex, which is absent in C. elegans, but of major importance in Drosophila and higher metazoans. PMID- 14560956 TI - Genetic analysis of TOR signaling in Drosophila. AB - Over a 4-day period of development, Drosophila larvae undergo a roughly 1,000 fold increase in mass. This impressive growth requires a continuous source of dietary protein; in the absence of amino acids, growth is arrested and various larval tissues display characteristic cell-cycle, metabolic, and structural changes. Mutations in the Drosophila target of rapamycin (dTOR) gene result in strikingly similar phenotypes, suggesting that dTOR acts in a signaling pathway responsive to nutrient availability. Genetic epistasis experiments indicate that dTOR is also required for cell growth in response to insulin and PI3K signaling, and that S6K activation can partially rescue dTOR loss of function. Thus dTOR has roles in both nutrient- and growth factor-mediated signaling, and may act to coordinate the activities of these pathways during development. Here we describe the use of mutations in dTOR to dissect its role in various signaling events, to gain insight into TOR protein structure, and to identify novel factors involved in TOR signaling. PMID- 14560957 TI - Interplay between growth factor and nutrient signaling: lessons from Drosophila TOR. AB - During normal development, cellular and organismal growth is coordinately regulated. Each cell and each individual organ integrates information about nutrient availability, hormonal signals, and intrinsic growth programs. Describing the signaling pathways involved in these processes and how they are integrated is important to understand how growth is controlled during development and may also permit the development of means to curb uncontrolled growth in disease. In recent years, the biochemical analysis of cellular growth in cultured cells and the genetic dissection of growth control in model organisms has identified two conserved signaling pathways dedicated to cellular growth. The target of rapamycin (TOR) pathway regulates growth in response to nutrients, and the insulin/IGF pathways are involved in coordinating cellular growth in response to endocrine signals. This review discusses recent advances in the understanding of the interaction between these pathways, with a special focus on the contribution of the genetic analysis of these pathways in Drosophila. PMID- 14560958 TI - mTOR signaling to translation. AB - Over the past few years, the target of rapamycin (TOR) pathway has been implicated in the control of translation, both in yeast and in higher eukaryotes. In this review, we provide an overview of translation in eukaryotes, and discuss the mechanisms and advantages of the regulation of translation. We then describe how the TOR pathway can modulate translation in yeast and in mammals, through the modulation of the phosphorylation of key translation components, and the regulation of the abundance of ribosomes and translation factors. PMID- 14560959 TI - Modulation of the protein kinase activity of mTOR. AB - mTOR is a founding member of a family of protein kinases having catalytic domains homologous to those in phosphatidylinositol 3-OH kinase. mTOR participates in the control by insulin of the phosphorylation of lipin, which is required for adipocyte differentiation, and the two translational regulators, p70S6K and PHAS I. The phosphorylation of mTOR, itself, is stimulated by insulin in Ser2448, a site that is also phosphorylated by protein kinase B (PKB) in vitro and in response to activation of PKB activity in vivo. Ser2448 is located in a short stretch of amino acids not found in the two TOR proteins in yeast. A mutant mTOR lacking this stretch exhibited increased activity, and binding of the antibody, mTAb-1, to this region markedly increased mTOR activity. In contrast, rapamycin FKBP12 inhibited mTOR activity towards both PHAS-I and p70S6K, although this complex inhibited the phosphorylation of some sites more than that of others. Mutating Ser2035 to Ile in the FKBP12-rapamycin binding domain rendered mTOR resistant to inhibition by rapamycin. Unexpectedly, this mutation markedly decreased the ability of mTOR to phosphorylate certain sites in both PHAS-I and p70S6K. The results support the hypotheses that rapamycin disrupts substrate recognition instead of directly inhibiting phosphotransferase activity and that mTOR activity in cells is controlled by the phosphorylation of an inhibitory regulatory domain containing the mTAb-1 epitope. PMID- 14560960 TI - Role of mTOR signalling in the control of translation initiation and elongation by nutrients. AB - Protein synthesis requires nutrients both as precursors (amino acids) and as a source of energy, since this process consumes a high proportion of cellular metabolic energy. Recent work has shown that both types of nutrients directly influence the activities of components of the translational machinery in mammalian cells. Amino acids positively regulate signalling through the mammalian target of the rapamycin (mTOR) pathway, although the degree of dependency on external amino acids varies between cell types. mTOR signalling modulates several key components involved in mRNA translation, in particular (via repressor proteins) the cap-binding initiation factor eIF4E, the ribosomal protein S6 kinases, and elongation factor eEF2. The branched-chain amino acid leucine is the most effective one in most cell types. It is currently unclear how mammalian cells sense prevailing amino acid levels, although this may involve intracellular amino acids. Cellular ATP levels can also influence mTOR activity. The activities of some translation factors are modulated by mTOR-independent mechanisms. Examples include the regulation of eEF2 by cellular energy levels, which may be controlled via the AMP-activated protein kinase, and the activity of the guanine nucleotide-exchange factor eIF2B, which is modulated by amino acids and metabolic fuels. PMID- 14560961 TI - Novel regulatory mechanisms of mTOR signaling. AB - As a master regulator of cellular processes ranging from cell growth and proliferation to differentiation, the mammalian target of rapamycin (mTOR) is critically involved in a complex signaling network. mTOR appears to govern an amino acid sensing pathway that integrates with a phosphatidylinositol 3-kinase dependent mitogenic pathway to activate the downstream effectors. Recent findings have revealed some unexpected regulatory mechanisms of mTOR signaling. A direct link between mTOR and mitogenic signals is found to be mediated by the lipid second messenger phosphatidic acid. In addition, cytoplasmic-nuclear shuttling of mTOR appears to be required for the cytoplasmic functions of this protein. A new picture of the rapamycin-sensitive signaling network is emerging, with implications in putative upstream regulators and additional downstream targets for mTOR. PMID- 14560962 TI - Raptor and mTOR: subunits of a nutrient-sensitive complex. AB - mTOR/RAFT1/FRAP is the target of the FKBP12-rapamycin complex as well as a central component of a nutrient- and hormone-sensitive pathway that controls cellular growth. Recent work reveals that mTOR interacts with a novel evolutionarily conserved protein that we named raptor, for "regulatory associated protein of mTOR." Raptor has several roles in the mTOR pathway. It is necessary for nutrient-mediated activation of the downstream effector S6K1 and increases in cell size. In addition, under conditions that repress the mTOR pathway, the association of raptor with mTOR is strengthened, leading to a decrease in mTOR kinase activity. Raptor is a critical component of the mTOR pathway that regulates cell growth in response to nutrient levels by associating with mTOR. PMID- 14560963 TI - Kinase activities associated with mTOR. AB - Although mTOR is a member of the PI-kinase-related kinase family, mTOR possesses serine-threonine protein kinase activities, which phosphorylate itself and exogenous substrates. mTOR autophosphorylates in vitro and is phosphorylated in vivo on serine residues. Ser2481, which is located in a His-Ser-Phe motif near the conserved carboxyl-terminal mTOR tail, has been reported as an autophosphorylation site in vivo and in vitro. The significance of the autophosphorylation remains unclear. Another phosphorylation site on mTOR in vivo is Ser2448. This site appears not to be an autophosphorylation site but a site potentially phosphorylated by protein kinase B (PKB). mTOR immunopurified from culture cells or tissues phosphorylates in vitro p70 S6 kinase (p70) alpha and p70beta, mainly on Thr412 or Thr401, respectively, located in a Phe-Thr-Tyr motif. Another exogenous substrate phosphorylated by immunopurified mTOR in vitro is eIF4E-binding protein 1 (4E-BP1) at sites corresponding to those phosphorylated in vivo during insulin stimulation in a Ser/Thr-Pro motif. Recently, raptor, a 150-kDa TOR-binding protein that contains a carboxyl-terminal WD-repeat domain, was discovered as a scaffold for the mTOR-catalyzed phosphorylation of 4E-BP1 and for the mTOR-mediated phosphorylation and activation of p70alpha. Other potential substrates phosphorylated by mTOR are nPKCdelta, nPKCepsilon, STAT3, and p53. The requirement of raptor for binding to and phosphorylation by mTOR of these potential substrates would clarify their physiological importance in the mTOR signaling pathway. PMID- 14560964 TI - mTOR: a mediator of intracellular homeostasis. AB - Earlier studies have shown that mTOR plays a key role in ribosome biogenesis. In bacteria, amino acids and ATP levels independently control ribosome biogenesis. Here, we describe recent findings demonstrating that homeostatic levels of amino acids, most notably branched-chain amino acids, and ATP, independently regulate the activity of mTOR. Unlike the effects of amino acids, the effects of ATP appear to be direct. Based on these findings we propose a model by which tumor cells existing in the anaerobic environment may have an advantage in growth by exploiting the rapid, although less efficient, production of ATP to drive growth via the mTOR signaling pathway. PMID- 14560965 TI - mTOR as a positive regulator of tumor cell responses to hypoxia. AB - Rapamycin is a clinically approved immunosuppressive agent that has recently shown promising antitumor activities in human patients. In contrast to many conventional chemotherapeutic agents, rapamycin displays a remarkably high level of selectivity for certain types of tumors. The pharmacological activities of rapamycin are attributable to the functional inhibition of a single target protein, termed the mammalian target of rapamycin (mTOR). Because mTOR is widely expressed in both normal and transformed cells, variations in mTOR expression levels are likely not a primary determinant of tumor sensitivity to rapamycin. However, recent studies highlighted an intriguing link between cancer cell sensitivity to rapamycin and deregulated signaling through the phosphoinositide (PI) 3-kinase pathway. These findings have prompted a search for cancer-related responses that are jointly regulated by the PI 3-kinase signaling cascade and mTOR. The oxygen-regulated transcription factor, hypoxia-induced factor (HIF)-1, has emerged as a candidate target for both of these two highly interactive signaling proteins. Here we review evidence that mTOR functions as a positive regulator of HIF-1-dependent responses to hypoxic stress in human cancer cells. PMID- 14560966 TI - Retroviral oncogenes and TOR. AB - Retroviruses have recruited the catalytic subunit of PI 3-kinase and its downstream target, Akt, as oncogenes. These viruses cause tumors in animals and induce oncogenic transformation in cell culture. The oncogenicity of these viruses is specifically inhibited by rapamycin; retroviruses carrying other oncogenes are insensitive to this macrolide antibiotic. Rapamycin is an inhibitor of the TOR (target of rapamycin) kinase whose downstream targets include p70 S6 kinase and the negative regulator of translation initiation 4E-BP. Emerging evidence suggests that the TOR signals transmitted to the translational machinery are essential for oncogenic transformation by the PI 3-kinase pathway. PMID- 14560967 TI - mTOR as a target for cancer therapy. AB - The target of rapamycin, mTOR, acts as a sensor for mitogenic stimuli, such as insulin-like growth factors and cellular nutritional status, regulating cellular growth and division. As many tumors are driven by autocrine or paracrine growth through the type-I insulin-like growth factor receptor, mTOR is potentially an attractive target for molecular-targeted treatment. Further, a rationale for anticipating tumor-selective activity based on transforming events frequently identified in malignant disease is becoming established. PMID- 14560968 TI - The endogenous development, described by light and electron microscopy, of Eimeria jamescooki sp. n. (Apicomplexa: Eimeriidae) from the skink Cryptoblepharus virgatus. AB - Eimeria jamescooki sp. n. was recovered from the skink Cryptoblepharus virgatus (Garman) found on the grounds of James Cook University, Townsville (type locality), North Queensland, Australia. Oocysts were 17.5-25.0 (22.1 +/- 1.9) x 15-22.5 (17.7 +/- 1.6) microm and sporocysts 6.25-10.0 (7.9 +/- 1.15) x 3.75-6.25 (5.3 +/- 1.0) microm in size. Endogenous stages are described from histological material examined by light microscope and by transmission electron microscope. Both merogony stages and gamonts were found to develop in the cytoplasm of the anterior gut mucosal epithelium. Meront progeny were comprised of 10 to 21 merozoites. Premature macrogamonts were elongate; some host cells contained two elongate macrogamonts. Unique to the presently described species were the Golgi "plaques" and an enclosure of tubuli. Mature macrogamonts and young oocysts ranged in size from 14 x 7 to 21 x 11 microm and contained two types of wall forming bodies, canaliculi and amylopectin granules. Differentiating microgamonts conformed in fine structure with that observed in other eimerians. Their sizes increased from 15.4 x 4.2 to 28 x 8.4 microm while dividing to over 70 nuclei, which formed a corresponding yield of microgametes. PMID- 14560969 TI - Genotypic, phenotypic, biochemical, physiological and pathogenicity-based categorisation of Acanthamoeba strains. AB - The genus Acanthamoeba includes more than 20 morphological species, but classification is problematical. Recently, the discovery of substantial interstrain differences in ribosomal DNA (rDNA) sequences has prompted questions about the relatedness of strains of the same species. In this study, therefore, we have investigated relationships between two isolates of A. polyphaga, CCAP 1501/3c and ATCC 30871, using morphological, biochemical, physiological, molecular and cytotoxicity assays. We observed that A. polyphaga ATCC 30871 exhibited up to six arms in endocyst while A. polyphaga CCAP 1501/3c exhibited a maximum of 5 arms thus indicating their position in group 2 and 3, respectively. Acanthamoeba polyphaga ATCC 30871 exhibited growth at 37 degrees C and growth on 1M mannitol plates while A. polyphaga CCAP 1501/3c did not. In addition, both isolates exhibited differences in isoenzyme banding patterns and rDNA restriction fragment polymorphisms. More importantly, A. polyphaga ATCC 30871 produced cytotoxicity on corneal epithelial cells while A. polyphaga CCAP 1501/3c had no effects, suggesting differences in pathogenicity. Thus, all the results provide evidence for significant differences between the strains and further provided the basis for reclassification of the isolates. Implications of these results in the clinical diagnosis of pathogenic Acanthamoeba are discussed. PMID- 14560970 TI - Acolpenteron australe sp. n. (Dactylogyridae:Dactylogyrinae), a new species from the ureters of Percichthys trucha (Perciformes:Percichthyidae) in Patagonia (Argentina). AB - Acolpenceron australe sp. n. (Dactylogyridae, Dactylogyrinae) is described from ureters and renal tubules of Percichthys trucha (Cuvier et Valenciennes) (Perciformes, Percichthyidae) from Andean Patagonian lakes. The new species has a haptor with 14 hooks, with shanks comprised of two subunits. It has overlapped intercaecal gonads, male copulatory organ as a sclerotized tube with one counterclockwise coil and a J-shaped accessory piece. It differs from the other species of Acolpenteron by having a non-forked accessory piece. This is the first monogenean species described from a percichthyid host in South America. PMID- 14560971 TI - Larval spirurida (Nematoda) from the crab Macrophthalmus hirtipes in New Zealand. AB - Previously undescribed third-stage larvae of two species of Spirurida were found in the haemocoel of the stalk-eyed mud crab Macrophthalmus hirtipes (Heller) (Ocypodidae) in New Zealand. Examinations by light and scanning electron microscopy showed that the larger larvae (about 7 mm long) belonged to a species of Ascarophis van Beneden, 1871 (Cystidicolidae), the genus including parasites of fishes, whereas the smaller larvae (about 4-5 mm long) belonged to the Acuariidae, a family with species parasitic as adults mostly in aquatic birds. In a sample of 82 specimens of M. hirtipes collected in July 2002 from Papanui Inlet, on Otago Peninsula, South Island, 74 crabs (90.2%) were infected with larval nematodes with an intensity of 1-18 (mean 4.6) nematodes per crab; no distinction between nematode species was made in these estimates, although juvenile Acuariidae greatly outnumbered larval Ascarophis. Apparently, crabs play a role as intermediate hosts of these nematode species. This is the first record of larval representatives of Cystidicolidae and Acuariidae from invertebrates in the Australasian Region. PMID- 14560972 TI - Redescription of Pterygodermatites (Mesopectines) nycticebi (Monnig, 1920) (Nematoda: Rictulariidae), a parasite of slow loris Nycticebus coucang (Mammalia: Primates). AB - Pterygodermaties (Mesopectines) nycticebi (Monnig, 1920) (Nematoda: Spirurida: Rictulariidae) is redescribed based on immature and mature adults collected from the stomach and small intestine at autopsy of a slow loris, Nycticebus coucang (Boddaert, 1785) (Mammalia: Primates), in a zoological garden in Japan. It is first demonstrated that male possesses a minute telamon and a left lateral pore in the preanal part of body. The cause of death of the slow loris is strongly surmised to be related to the nematode infection, which was apparently acquired under captivity in the zoological garden. PMID- 14560973 TI - Systematic status of Manaosia bracodemoca Woodland, 1935 and Paramonticellia itaipuensis Pavanelli et Rego, 1991 (Eucestoda: Proteocephalidea), parasites of Sorubim lima (Siluriformes: Pimelodidae) from South America. AB - Type material of the proteocephalidean cestodes Manaosia bracodemoca Woodland, 1935 and Paramonticellia itaipuensis Pavanelli et Rego, 1991 (both monotypic genera) as well as recently collected material of the latter species are redescribed. A close similarity between both species was observed: the shape of scoleces is identical, both possess a globular scolex with hidden suckers, a well developed circular, horseshoe-shaped musculature surrounding suckers; the measurements of both taxa are also similar (scolex, suckers, young proglottides). Both cestodes possess a medullary ovary crossing the dorsal muscle layer and ending in the dorsal cortex, the testes are in the dorsal cortex and the cortical vitellarium is laterally situated. Both species parasitize the same fish host. Therefore Paramonticellia is considered a junior synonym of Manaosia and Paramonticellia itaipuensis becomes a junior synonym of Manaosia bracodemoca. An emended diagnosis of Manaosia, which is a monotypic genus, is provided. PMID- 14560974 TI - Activity levels and predator detection by amphipods infected with an acanthocephalan parasite, Pomphorhynchus laevis. AB - The acanthocephalan parasite Pomphorhynchus laevis (Muller, 1776) uses freshwater amphipods as its intermediate host. In order to complete the life cycle, the infected amphipod must be consumed by a fish, where the acanthocephalan will mature and reproduce. Parasite transmission, and therefore fitness, could be enhanced if infected amphipods fail to detect or avoid predatory fish. We compared the activity levels of infected and non-infected amphipods, Echinogammarus stammeri (Karaman, 1931), in both the presence and absence of odours from its natural, definitive host, the fish Leuciscus cephalus (L.). Throughout the experiment, infected amphipods were more active than were non infected individuals. The non-infected amphipods reduced their activity after the addition of fish odours, but the infected amphipods failed to show a significant decrease. The failure of infected amphipods to reduce activity levels in the presence of fish odour may reflect a parasite strategy to increase its chances of transmission by making its amphipod host more vulnerable to predation by fish. PMID- 14560975 TI - Life cycle of the temporary fish parasite, Gnathia africana (Crustacea: Isopoda: Gnathiidae). AB - Laboratory work was conducted to elucidate the life cycle of the South African gnathiid isopod, Gnathia africana Barnard, 1914. The natural fish hosts of this temporary parasite, the super klipfish Clinus superciliosus (Linnaeus, 1758), were exposed to gnathiid larvae in the laboratory. It was found that G. africana has three larval stages, consisting of three unfed (zuphea) and three fed (praniza) stages. First-, second- and third-stage zuphea larvae took an average of 2 h 18 min, 2 h 43 min and 10 h 8 min respectively to complete their feeding and the first- and second-stage praniza moulted at 8 and 10 days respectively into the next zuphea stage. Three to six days after its last blood meal, the sex of the third and final praniza stage could be determined by the presence of either a testis or two ovaries in the dorsal pereon. Male larvae moulted into adult males between 8 and 10 days post feeding. Female larvae moulted at approximately 17 days into adult females. Fertilisation of the eggs by the male took place within 24 hours of completion of the female moult. The development of the embryos and subsequent release of the young larvae between 15 and 23 days post fertilisation completed the cycle. This entire cycle took approximately 62 days in water temperatures of 20-25 degrees C. PMID- 14560976 TI - Chigger mites (Acari: Trombiculidae) new to the fauna of Cuba, with the description of two new species. AB - Two new species of chigger mites, Hyponeoculta monocoxalae sp. n. from bats and reptiles, and Perares nudosetosus sp. n. from bats, are described. The first finding of larvae of Tectumpilosum negreai Feider, 1983 in nature is reported from a bat collected at the type locality, and the description of this species is emended. Four species, Perates monops (Brennan et Jones, 1960), Parasecia manueli (Brennan et Jones, 1960), Beamerella acutascuta Brennan, 1958, and Blankaartia sinnamaryi (Floch et Fauran, 1956), are recorded for the first time in Cuba and on new host species. PMID- 14560977 TI - Henneguya curvata sp. n. (Myxosporea: Myxobolidae) parasitizing the gills of Serrasalmus spilopleura (Characidae: Serrasalminae), a South American freshwater fish. PMID- 14560978 TI - On the presence of Lecithochirium musculus (Digenea: Hemiuridae) in Conger conger. PMID- 14560979 TI - Co-occurrence of metacestodes of Echinococcus multilocularis and Taenia taeniaeformis (Cestoda) in Arvicola terrestris (Rodentia) in France. PMID- 14560980 TI - Effect of the salivary gland and midgut extracts from Ixodes ricinus and Dermacentor reticulatus (Acari: Ixodidae) on the growth of Borrelia garinii in vitro. PMID- 14560981 TI - Low plasma protein Z levels in patients with ischemic colitis. AB - Hypercoagulable states have been suggested to play an important role in the pathogenesis of ischemic colitis. Since protein Z is, as recently demonstrated, important in the regulation of coagulation, we investigated the plasma levels of protein Z in connection to factor V Leiden (FVL) and anti-phospholipid antibodies in patients with a definite diagnosis of ischemic colitis. The plasma levels of protein Z were measured using a commercially available enzyme-linked immunosorbent assay in 33 patients with ischemic colitis, 13 patients with diverticulitis, and 33 healthy controls. Mean plasma protein Z levels were 1.38 +/- 0.52 microg/ml in patients with ischemic colitis and were significantly lower compared to healthy controls (1.86 +/- 0.49 microg/ml) and patients with diverticulitis (1.72 +/- 0.53 microg/ml) (P = 0.001). Protein Z deficiency was found in patients cases with ischemic colitis (18.2%) compared to one with diverticulitis (7.7%) and one healthy control (3.0%). In conclusion, our results suggest that low plasma protein Z levels may play a role in the disease development in some cases with ischemic colitis. PMID- 14560982 TI - Effects of intestinal ischemia-reperfusion injury on rat peripheral blood neutrophil activation. AB - Peripheral blood neutrophil activation status is indicative of remote organ damage after intestinal ischemia secondary to aortic aneurysm repair. However, the effects of direct intestinal ischemia-reperfusion (I/R) injury on neutrophil activation and its reflection of remote organ injury have not been evaluated. DA rats were subjected to 30 min of intestinal ischemia or sham surgery. Blood samples were taken before ischemia and 30, 60, and 120 min after reperfusion. Neutrophil counts were quantified and CD11b, CD62L, and NKR-P1 expression was assessed using flow cytometry. The sham procedure induced increases in neutrophil numbers (P < 0.001), which was transiently attenuated in animals subjected to intestinal I/R injury. CD11b expression increased in both groups, whereas CD62L and NKR-P1 (P < 0.01) expression decreased in both groups. These findings suggest that even mild surgical procedures induce demargination of neutrophils. Monitoring the peripheral blood for activated neutrophils is of no value in assessing the severity of direct intestinal I/R injury or predicting remote organ damage after intestinal ischemia. PMID- 14560983 TI - Small bowel obstruction caused by metastatic squamous cell carcinoma of the skin without lymphadenopathy in a renal transplant recipient. PMID- 14560984 TI - High prevalence of steatorrhea in 101 diabetic patients likely to suffer from exocrine pancreatic insufficiency according to low fecal elastase 1 concentrations: a prospective multicenter study. AB - Impaired exocrine pancreatic secretion has been frequently observed in diabetic patients by different methods, including direct function tests. However, the clinical importance remained unclear. In the present study, the fecal fat excretion in patients with type 1 or type 2 diabetes mellitus and exocrine dysfunction according to fecal elastase 1 concentrations <100 microg/g was investigated. Subjects with a history of gastrointestinal cancer, gastrointestinal surgery, alcohol abuse, or inflammatory diseases were excluded. In 101 patients the mean (+/- SD) fat excretion was 9.19 +/- 5.39 g. Only 41 patients (40.6%) had normal fat excretion <7 g/day. In 40 patients (39.6%), it was higher than 10 g/day, indicating relevant steatorrhea. The fat excretion did not correlate with diabetes type, duration, or clinical symptoms. This finding is of some clinical importance and might influence pathophysiological concepts and the management of diabetic patients. PMID- 14560986 TI - Pancreatic ascites in an infant: lack of symptoms and normal amylase. AB - A 4-month-old boy presented with 9 days of abdominal distension. The abdomen was tense, distended, and nontender, with a fluid wave. Hypoalbuminemia, hyponatremia, high lipase, normal amylase, high ascitic fluid: lipase, amylase, and serum-ascites albumin gradient < 1.1 were present. Abdominal CT showed large ascites, edema, and pancreatic cyst. No improvement was noted with bowel rest, TPN, albumin, furosemide, octreotide, and paracentesis. Endoscopic retrograde cholangiopancreatography showed disrupted pancreatic duct and a cyst. Pancreatic duct stenting was complicated by early outward migration of the stent and was thus ineffective. An exploratory laporatomy revealed a cyst. Cystogastrostomy resolved the pancreatitis and ascites. The patient was discharged off TPN and tolerating enteral nutrition. Pancreatic ascites is rare, producing few or no symptoms in infants. In conclusion, our patient may have had viral pancreatitis, complicated by a disrupted duct and/or ruptured pseudocyst with ascites formation. Medical management was ineffective. Surgery appears to have been curative. PMID- 14560985 TI - Role of duodenum on sphincter of Oddi motility in conscious dogs. AB - Our aim was to determine the role of the duodenum in controlling sphincter of Oddi motility using conscious dogs after total duodenectomy. In a control group (N = 6), a cannula was implanted into the duodenum opposite to the papilla to allow retrograde sphincter manometry. In a duodenectomy group (N = 6), the papillae were preserved at total duodenectomy and sutured to thejejunum anastomosed to the stomach (neoduodenum). The cannula was implanted opposite to the implanted papillae. Interdigestive and postprandial sphincter and duodental or neoduodenal motility were recorded by manometric and myoelectric methods. Duodenectomy disrupted sphincter cyclic motility associated with the intestinal migrating motor complex and increased sphincter activity throughout the cycle. Sphincter activity increased immediately after feeding and did not differ between the two groups. In conclusion, during the interdigestive period, the duodenum has a distinct role in regulating sphincter cyclic motility. The initiation of the fed pattern of sphincter motility does not need the duodenum. PMID- 14560987 TI - A case of autoimmune pancreatitis with initially negative autoantibodies turning positive during the clinical course. PMID- 14560988 TI - Dysphagia and advancing age: are manometric abnormalities more common in older patients? AB - The universal process of aging may result in physiologic deterioration. Dysphagia may be more common in older patients. The effect of aging on esophageal manometry is not well established. The aim of this study was to determine if esophageal motility studies and associated symptoms in older patients with dysphagia differ significantly from younger patients. Patients who were 65 years of age or older (N = 53) were compared with patients who were 18-45 years of age (N = 53). Presenting symptoms, manometric findings, and diagnoses were compared between the two groups. In the older group, there were 29 women (55%), in the younger group there were 35 women (66%). The mean age of the older group was 75 +/- 7 years, the mean age in the younger group was 34 +/- 7 years. All patients reported dysphagia to solids. No significant differences were found in the reporting of associated symptoms. There were no significant differences in average lower esophageal sphincter (LES) resting pressure, residual LES pressure, LES relaxation, or peristalsis between groups. Older patients were as likely to have a normal study as younger patients (18% vs 23%, P = NS) and were also as likely to have the diagnosis of achalasia (32% vs 34%, P = NS). In conclusion, older and younger patients referred for manometric study of dysphagia have similar manometric findings. Esophageal manometry can be helpful in determining abnormalities in motility in both older and younger patients. PMID- 14560989 TI - Effect of preinjury large bowel emptying on the inhibition of upper gastrointestinal motility after spinal cord injury in rats. AB - Spinal cord transection (SCT) inhibits gastrointestinal motility in rats. We evaluated the effect of preinjury large bowel emptying on this phenomenon. Male Wistar rats (N = 52) were fasted for 24 or 48 hr with water ad libitum and pretreated with lactose (0.8 g) or saline. Next, laminectomy followed or not by complete SCT between T4 and T5 vertebrae was performed. Phenol red recovery in the stomach and proximal, medial, and distal small intestine was determined 1 day later. In animals submitted to 24 hr fasting + saline, SCT increased gastric recovery by 42.8% and decreased medial small intestine recovery by 56.2%, while 48 hr fasting + saline or 24 hr fasting + lactose prevented the inhibition of gastric emptying (GE) in SCT animals. The 48 hr fasting + lactose prevented the inhibition of both GE and gastrointestinal transit. SCT-induced inhibition of upper gastrointestinal motility may involve enhancement of inhibitory reflexes, which can be prevented by large bowel emptying. PMID- 14560990 TI - Selection of patients with gastroesophageal reflux disease for antireflux surgery based on esophageal manometry. AB - Gastroesophageal reflux disease (GERD) is caused by a mechanically defective lower esophageal sphincter (LES) and may be worsened by impaired esophageal peristalsis. The aim of this study was to evaluate the efficacy of medical treatment depending on the function of the LES and esophageal peristalsis. We studied 128 GERD patients with mild esophagitis. Group 1 (N = 26) consisted of patients with a normal LES and normal esophageal peristalsis. Group 2 (N = 63) comprised patients with a defective LES but normal peristalsis. Patients of group 3 (N = 39) had a defective LES as well as impaired esophageal peristalsis. The patients were continuously treated with omeprazole. Clinical evaluation and endoscopy were repeated after 3, 6, and 12 months. Recurrence of GERD was diagnosed if there was relapse of heartburn and/or esophagitis. The recurrence rate was 7.7% in group 1, 38.1% in group 2 (P < 0.05) and 79.5% in group 3 (P < 0.05). In conclusion, in GERD patients with a mechanically defective LES, especially in those with deteriorated esophageal peristalsis, antireflux surgery should be considered since medical therapy reveals a high recurrence rate. PMID- 14560991 TI - Gastroesophageal reflux disease: review of presenting symptoms, evaluation, management, and outcome in infants. AB - We conducted a retrospective review of 342 infants presenting with symptoms suggestive of gastroesophageal reflux disease (GERD) to assess their evaluation, management, and outcome. All infants underwent extended pH monitoring (EPM) for one or more of the following symptoms: regurgitation, choking, irritability, failure to thrive, apparent life-threatening event, or wheezing. EPM was considered abnormal if distal reflux index was > or = 5% and/or if the Euler and Byrne score was > or = 50 and these patients were labeled as having GERD. Those with normal EPM parameters were controls. All infants were < 1 year of age (198 boys, 144 girls). Most received conservative treatment with formula changes and/or thickening and positional management before EPM. Of 342 infants studied, EPM was normal in 169 patients, ie, controls, and GERD was present in 173 children. GERD was more prevalent in Caucasians compared to African Americans (P < 0.045). Although a prokinetic or acid suppression medication or both were given more commonly in infants with GERD, almost half of infants in the control group also needed pharmacotherapy because of persistent symptoms. Mean time to resolution of symptoms in the control group infants with normal EPM was 3.5 months, and in infants with GERD it was 5.4 months (P < 0.001). In conclusion, the majority of infants with symptoms suggestive of GERD resolve their symptoms within 3-6 months of initiating treatment. Day-to-day variability of the spectrum of GERD may contribute towards normal results of EPM in some infants who need pharmacotherapy. PMID- 14560992 TI - Does intragastric nitrite concentration reflect gastric carcinogenesis in Japanese Helicobacter pylori-infected patients? AB - Established risk factors for gastric cancer include a diet high in nitrate or nitrite and low in vitamin C and the presence of achlorhydria or hypochlorhydria. The aim of this study was to investigate the relationship between intragastric nitrite concentration and atrophic change of the stomach or gastric carcinogenesis in Japanese Helicobacter pylori-infected patients. Gastric juice pH, nitrite, and total vitamin C concentrations in gastric juice, serum pepsinogen I and II concentrations, and specific Helicobacter pylori antibody were analyzed. Intragastric total vitamin C concentration was decreased by Helicobacter pylori infection of the gastric mucosa and with progression of the atrophic grade. There was a significant positive correlation between atrophic grade and intragastric nitrite concentration. In conclusion, the levels of nitrite in gastric juice play a causal role in the development of cancer in Helicobacter pylori-associated atrophic gastric mucosa. PMID- 14560993 TI - Human colon carcinomas constitutively express and shed type II IL-1 receptor, an IL-1 antagonist. AB - We reported earlier that rat intestinal epithelial cells respond to helminth infection, to NSAID injury, and to detachment in vitro with expression of the IL 1RII. Now we have sought to determine whether human colon carcinoma cell lines express, or may be induced to express, this potent IL-1 antagonist. Using RT-PCR, the T84 and HT-29 cell lines constitutively expressed mRNA for the membrane bound, but not the secreted variant of the receptor. The protein was detectable by immunohistochemistry and was estimated to be 70 kDa by western blotting. TNF treatment of T84 cells led to slightly increased levels of IL-1RII mRNA and to significant increases in soluble protein detected in culture supernatants. Treating T84 cells with inhibitory anti-IL-1RII antibodies led to heightened responsiveness to IL-1, measured as IL-8 production. Expression of the IL-1RII by human epithelial cells has implications in terms of the IL-1 agonist versus antagonist balance in the diseased intestines. PMID- 14560994 TI - Glucocorticoid receptor (GR) immunohistochemical expression is correlated with cell cycle-related molecules in human colon cancer. AB - The aim of this study was to examine glucocorticoid receptor (GR) immunohistochemical expression in colon cancer histopathological specimens and to correlate it with clinicopathological parameters, tumor proliferative capacity, cell cycle-related molecule expression, and patients' survival. Primary tumoral samples from 91 colon cancer patients were immunostained for the detection of GR, cyclins D1 and E, Rb protein (pRb), p16, p21, and Ki-67, using the streptavidin biotin-peroxidase technique. GR expression was correlated with tumor histopathological characteristics and proliferative capacity, cell cycle-related molecule expression, and patients' survival. GR positivity was prominent in 44 of 91 (48%) colon cancer cases and was positively correlated with the expression of cell cycle-related molecules pRb (P = 0.008) and p16 (P = 0.002), while lack of correlation was noted with cyclins D1 and E and p21. GR expression was not correlated with tumor location, grade of differentiation, Dukes' stage, lymph node and liver metastasis, venous invasion, tumor proliferative capacity (evident by Ki-67-labeling status) and patient survival. Our findings support evidence for GR participation in the biological mechanisms underlying the carcinogenic evolution in the colon, implying the use of glucocorticoids as an adjuvant treatment for cell cycle modulation in colon cancer cells. PMID- 14560996 TI - Vascular pancreatic gastric fistula: a complication of colloidal 32P injection for nonresectable pancreatic cancer. PMID- 14560995 TI - Fecal primary bile acids and serum cholesterol are associated with colorectal adenomas. AB - In order to identify biomarkers of colorectal tumors, 20 subjects with colorectal adenomas were compared with 20 controls as regards fecal parameters (pH, short chain fatty acids, bile acids, and sterols), blood parameters (bile acids, cholesterol, triglycerides, glycemia and insulinemia), and rectal cell proliferation. Variables were compared by unconditional logistic regression, controlling for gender. There were significant and positive associations between risk of adenoma and total fecal primary bile acids and serum cholesterol, with odds ratios for the third versus first tertile = 9.4 (P for trend = 0.03) and 8.6 (P for trend = 0.04), respectively. There was a trend towards an increased triglycerides level in adenoma subjects compared with controls (P = 0.08). These three parameters correlated with cell proliferation, although cell proliferation itself was not significantly associated with adenomas. In conclusion, these results suggest that fecal primary bile acids and serum cholesterol are markers of early events of colorectal carcinogenesis. PMID- 14560997 TI - A case of double carcinoid tumors of the gallbladder. PMID- 14560998 TI - Colonic hydrogen elimination and methane production in infants with and without infantile colic syndrome. AB - Our objective was to investigate the relationship between demographic factors, nutrition, stool gas production, and the existence of infantile colic (IC) syndrome. Hydrogen and methane production from stool specimens of infants with and without infantile colic was quantified at two separate time points, the age at presentation of colic (<12 weeks) and at >6 months of age. The relationship between demographic variables and IC was also studied. A total of 59 infants with ages ranging from 2 to 12 weeks were enrolled in the study. Of these, 30 infants developed symptoms of colic. No correlation was found between IC and birth weight, gestational age, sex, type of feeding, mean time of feeding, stool frequency, and consistency. There was also no correlation between IC and the parents' age or education or the infant's number of siblings. Analysis of the stool samples revealed that methane was produced at concentrations >2 ppm by 15.3% of the infants at age <3 months and by 46.4% of infants at age >6 months. The mean methane concentrations produced by stool increased with age (0.95 +/- 0.58 ppm at 3 months of age vs 1.29 +/- 0.65 ppm at 6 months of age. There was no difference in stool hydrogen concentration between infants with and without IC. In contrast, the mean methane level at 3 and 6 months of age was higher in infants without IC than with IC, but reached statistical significance only at 6 months of age (0.97 +/- 0.68 vs 0.93 +/- 0.46) (NS) at 3 months of age, and 1.56 +/- 0.55 vs 0.93 +/- 0.62 (P < 0.05) at 6 months of age respectively. Furthermore, infants that produced higher methane levels at 3 and 6 months of age had significantly (p < 0.05) less colic in the first months of life. In conclusions, methane production may play a role in the alleviation of IC. Future studies are needed to confirm our findings. PMID- 14560999 TI - Oxygen consumption and chloride secretion in rat distal colon isolated mucosa. AB - The aerobic metabolic cost of chloride secretion was studied in rat distal colon isolated mucosa under several conditions by simultaneous measurement of short circuit current and oxygen consumption under conditions that preserve vectorial ion transport. A low-chloride solution and the presence of bumetanide plus diphenylamine-2-carboxylate reduced short-circuit current by 75% and oxygen consumption by 25%. Ouabain decreased short-circuit current by 93% and oxygen consumption by 32%. Serotonin increased both variables by 59% and 33%, respectively. Bumetanide and diphenylamine-2-carboxylate reduced but did not abolish the effect of serotonin on short-circuit current and oxygen consumption. Changes in short-circuit current and oxygen consumption were linearly correlated under all conditions tested. It is concluded that, in the unstimulated rat distal colon epithelium, chloride secretion accounts for about 75% of ouabain-sensitive short-circuit current and oxygen consumption. Stimulated chloride secretion may demand over 40% of total oxygen consumption. PMID- 14561001 TI - Systemic vasculitis presenting with acute gastrointestinal hemorrhage: case report and review of the literature. PMID- 14561000 TI - Regional brain activation in response to rectal distension in patients with irritable bowel syndrome and the effect of a history of abuse. AB - Previous studies have demonstrated alterations in brain response to rectal distension in patients with irritable bowel syndrome (IBS) compared to controls. Our aim was to compare regional brain activity in response to rectal balloon distension in patients with IBS and healthy controls. We studied six patients with IBS and six healthy controls. Positron emission tomography scans were obtained during rectal balloon distensions. Statistical parametric mapping and region of interest analysis were performed to identify and compare differences in regional cerebral blood flow (CBF) for each distension pressure within and between the groups of interest. In post-hoc analyses, patients with a history of sexual or physical abuse were compared to patients without abuse. In response to rectal distension, controls exhibit a greater increase in anterior cingulate cortex (ACC) activity compared to the IBS group (Z = 3.2, P = 0.001). Thalamic activity was higher in the IBS patients relative to the control group (Z = 3.3, P < 0.001). Increased ACC activity was observed in IBS patients with no history of abuse (Z = 5.2, P < 0.001) similar to controls, whereas no such increased activity was noticed in the abused group. In conclusion, this study replicates previous findings showing alterations in brain response to rectal distension in patients with IBS. The observations on the effect of abuse suggest a possible modulating role of abuse history on this brain response. PMID- 14561002 TI - Significance of prior hepatitis B virus infection in the development of hepatocellular carcinoma in patients with chronic hepatitis C. AB - To clarify the clinical significance of prior hepatitis B virus (HBV) infection in the development of C-viral hepatocellular carcinoma (HCC), we conducted two studies: (1) Two hundred thirty-four patients with C-viral HCC and 320 patients with C-viral chronic liver disease without HCC admitted to our hospital between 1990 and 1994 were analyzed for the association of hepatitis B core antibody (HBcAb) positivity with HCC by multivariate logistic regression analysis, and this revealed HBcAb positivity as an independent risk factor for development of HCC adjusted for age and sex. (2) Four hundred fifty-nine patients with biopsy proven hepatitis C virus-related chronic liver disease between 1986 and 1998 were enrolled in the cohort study and followed for the development of HCC. During an average follow-up of 6.6 +/- 3.3 years, HCC developed in 63 patients, 37 of 160 patients positive for HBcAb and 26 of 299 patients negative for HBcAb. Multivariate Cox proportional regression analysis showed that the incidence of HCC increased by age, advanced stage of liver fibrosis, mean alanine aminotransferase value of more than 80 IU/liter, and positivity of HBcAb. Sustained virological responders after interferon therapy revealed a reduced risk for HCC development. In conclusion, prior HBV infection was shown to be one of the independent risk factors for development of HCC in C-viral chronic liver disease. PMID- 14561003 TI - Serum levels of alpha1-antitrypsin predict phenotypic expression of the alpha1 antitrypsin gene. AB - We conducted a retrospective analysis to determine if both alpha1-antitrypsin serum level and phenotype need be studied when evaluating children for alpha1-AT deficiency. We collected data from patients less than 19 years old who had both serum alpha1-AT level and phenotype determined over a 9-year period (January 1992 December 2000). Eighty-eight patients were identified and 15 had the PiZZ phenotype. The serum alpha1-AT level was below normal (normal 85-215 mg/dl) in all 15 PiZZ patients. Seventy-two of 73 non-PiZZ patients had normal or above normal serum levels. The sensitivity of the serum alpha1-AT level was 100%, and the specificity was 99%. The serum alpha1-AT level had a positive predictive value of 94% and a negative predictive value of 100%. We conclude that serum alpha1-AT levels are highly predictive of the PiZZ phenotype. Determination of the serum alpha1-AT level alone should be the initial test when evaluating for alpha1-AT deficiency. PMID- 14561004 TI - Granulocyte-colony stimulating factor administration ameliorates liver regeneration in animal model of fulminant hepatic failure and encephalopathy. AB - It has previously been shown that recombinant granulocyte-colony stimulating factor (rG-CSF) accelerates and enhances hepatocyte proliferation in partially hepatectomized rats. In the present study, we examined the effect of rG-CSF administration on liver injury, regeneration, and survival outcome in an experimental rat model of fulminant hepatic failure (FHF) and encephalopathy induced by repeated injections of thioacetamide (TAA). FHF was induced in adult male Wistar rats by three consecutive intraperitoneal injections of TAA, at intervals of 24 hr. The animals were also injected with either saline or rG-CSF. Serum biochemical parameters and blood ammonia levels, liver histology, stage of hepatic encephalopathy, and survival were statistically significantly improved in TAA-intoxicated and rG-CSF-treated rats compared to TAA-intoxicated and saline treated ones. Furthermore, rG-CSF not only ameliorated the histologically evident liver injury in a statistically significant manner but also enhanced the proliferative capacity of the hepatocytes. Our data confirm the beneficial effect of rG-CSF administration in this animal model of FHF and encephalopathy, supporting evidence for a possible use of rG-CSF as supportive therapy in the management of FHF. PMID- 14561005 TI - Fatal cholestatic liver failure associated with gemcitabine therapy. AB - Drug-induced hepatotoxicity accounts for more than a third of the cases of acute liver failure in the United States. In complex medical conditions, the diagnosis of drug-induced liver injury may be confounding and, specifically, the potential hepatotoxicity of chemotherapeutic agents may be easily overlooked. Two fatal cases of cholestatic hepatotoxicity have been previously reported, clearly implicating gemcitabine therapy. We report a third fatal case of cholestatic liver failure that we think is strongly linked to the use of gemcitabine. This chemotherapeutic agent is a fluorine analog with broad-spectrum antitumor activity commonly used in the treatment of breast, lung, prostate, and cervical cancer. The case we report is of a 45-year-old woman with a history of metastatic breast cancer to her spine. The patient was in remission for two years before she presented with a compensated mixed hepatitis of mild to moderate severity. Inpatient work-up found metastases to the right humerus and inferior pubic ramus, but none in the liver. Gemcitabine and carboplatin therapy was initiated for relapse of breast cancer. The patient's liver enzyme elevation diminished, but did not normalize before the start of chemotherapy. She received four courses of gemcitabine/carboplatin and subsequently presented with decompensated, severe cholestatic hepatitis. Transjugular liver biopsy displayed marked cholestasis and hepatocellular injury consistent with drug-induced hepatoxicity. Gemcitabine has been extensively studied in the oncology literature and at this time is thought to be a low-risk hepatotoxin causing hepatic adaptation and transient, reversible liver enzyme elevation, rarely leading to termination of gemcitabine therapy for solid tumors. We believe that gemcitabine therapy, particularly in the setting of preexisting liver injury or metastases to the liver, increases the relative risk of severe and potentially fatal hepatic injury possibly by idiosyncratic and dose dependent mechanisms. We recommend careful monitoring and dose adjustment of gemcitabine in patients with abnormal liver function tests or evidence of hepatic metastases until further study clarifies this issue. PMID- 14561006 TI - Successful obliteration of duodenal varices after endoscopic ligation. PMID- 14561007 TI - Comparison of a salivary/sputum pepsin assay with 24-hour esophageal pH monitoring for detection of gastric reflux into the proximal esophagus, oropharynx, and lung. AB - The purpose of this study was to determine whether measurement of salivary/sputum pepsin could be used as a surrogate marker for detecting gastroesophageal reflux using 24-hr esophageal pH monitoring as the gold standard. Patients with gastroesophageal reflux symptoms underwent simultaneous 24-hr esophageal pH monitoring and collection of saliva and sputum samples for pepsin measurement using a recently developed assay. In all, 16 patients provided 19 positive (10.6%) and 161 negative pepsin assays. The mean pH values for the positive pepsin samples were lower then the negative samples at both the proximal [5.34 (95% CI, 4.94-5.75) vs 6.12 (95% CI, 6.03-6.20; P < 0.01)] and distal [4.97 (95% CI, 4.61-5.33) vs 6.03 (95% CI, 5.92-6.15; P < 0.01)] pH probes. Proximal esophageal reflux was not detected in patients who had a negative pepsin assay (N = 12); in contrast, proximal esophageal reflux was documented in three of four patients with a positive assay. In conclusion, detection of pepsin in the saliva and/or sputum may provide a noninvasive method to test for the proximal reflux of gastric contents. PMID- 14561008 TI - Circadian variation of gastroesophageal reflux in children. AB - Asymptomatic infants and infants with gastroesophageal reflux have been noted to have peak reflux in the midafternoon. The aim of this study was to examine circadian variation of gastroesophageal reflux in children with both normal and abnormal 24-hr esophageal pH probe studies. Eighty-seven patients were studied and 15 of these patients were found to have a reflux index >4% for the duration of the study. These 15 patients were found to have significant circadian variation. Peak reflux indices (mean 12.60%) were detected in the mid-afternoon, with a smaller peak at night (mean 8.07%). Trough indices (mean 2.60%) occurred in the early morning. The 72 patients with normal studies were found to have less circadian variation, although peak reflux indices (mean 1.76%) occurred in the early morning. Clinicians should target their antireflux therapy to periods of peak gastroesophageal reflux. PMID- 14561009 TI - Hiatus hernia and intrathoracic migration of esophagogastric junction in gastroesophageal reflux disease. AB - The prevalence and clinical presentation of reducible and irreducible hiatus hernia were investigated within a gastro-esophageal reflux disease patient population. Reflux symptoms and esophagitis data were collected on 791 patients. The barium swallow was used to assess the esophagogastric junction. Clinical and endoscopic findings were tested to predict radiographic findings. The esophagogastric junction was normal in 17% of patients, 53% had a sliding hiatus hernia with a reducible esophagogastric junction; in 23% it was irreducible although axial, and 8% had massive incarcerated hiatus hernia. The presence of reducible sliding hiatus hernia did not influence clinical presentation. Axial irreducibility presented with long-standing severe symptoms and esophagitis in 80% of cases. Clinical and endoscopic findings predicted axial irreducibility in 52% of cases. In conclusion, sliding hiatus hernia with an reducible esophagogastric junction does not influence the severity of gastroesophageal reflux disease. An irreducible esophagogastric junction is associated with long standing severe gastroesophageal reflux disease. Clinical and endoscopic findings may only be indicative of axial esophagogastric junction irreducibility; thus barium swallow should be part of the work-up. PMID- 14561011 TI - Relationship between pepsinogen I&II and H. pylori infection considered with grade of atrophy and gastroduodenal diseases. AB - There are some reports the gastric atrophy induces the downregulation of PG I and, contrariwise, that H. pylori (Hp) infection induces PG I. The aim of this study is to clarify this discrepancy and the relationship between Hp infection and PG I&II. Biopsy specimens and sera were obtained from 207 patients with dyspeptic symptoms undergoing diagnostic endoscopy for Hp infection, atrophic pattern, and estimation of anti-Hp antibodies (Ab) and PG I&II. PG I&II levels were compared before and after eradication in both successful and unsuccessful cases in 53 peptic ulcer patients. In all, 164 cases were Hp positive and 27 cases were Hp negative. For atrophic pattern, the 27 Hp-negative patients were classified into the C1 (N = 26) and C2 patterns (N = 1), and the 164 Hp positive patients were classified into C1 (N = 27), C2 (N = 34), C3 (N = 28), O1 (N = 41), O2 (N = 22), and O3 patterns (N = 12). PG I&II decreased according to the atrophic grade among Hp-positive patients. There are substantial associations between Hp status and PG I&II levels in the C1 pattern, and PG I&II levels are statistically lower in successful cases after eradication. In conclusion, Up regulation of PG I&II is strongly associated with Hp infection and may lead to ulcer formation. PMID- 14561010 TI - Regulation of Na/H exchanger-1 in gastroesophageal reflux disease: possible interaction of histamine receptor. AB - Na/H exchanger-1 (NHE-1) isoform regulates intracellular pH and protects the esophageal mucosa. These functions are compromised in gastroesophageal reflux disease (GERD); however, the role and the underlying mechanism of NHE-1 regulation are obscure. To address this issue, NHE-1 protein and mRNA levels were measured by ECL western blot analysis and a semiquantitative RT-PCR using alpha actin as an internal control. Ouabain-sensitive and K-stimulated p nitrophenylphosphatase activity was measured as a marker of the sodium pump. The level of NHE-1 protein and mRNA and sodium pump activity was increased in GERD patients with or without esophagitis. Interestingly, myeloperoxidase (MPO) activity and infiltration of inflammatory cells were significantly increased in the GERD patients with esophagitis as compared to the GERD without esophagitis and the normal controls. This induction of NHE-1 and sodium pump was reversed in the GERD patients taking an H2 blocker, but not in those taking antacids. The internal control alpha-actin did not change under these conditions. Yield of total RNA and crude microsomal proteins was also statistically similar in all the test groups. These findings demonstrate that induction of NHE-1 is regulated posttranscriptionally through a possible interaction of histamine receptor. Induction of NHE-1 and sodium pump activity together might be a mechanism underlying the GERD pathogenesis and suggests a significance of NHE-1 inhibition in the treatment of GERD. PMID- 14561012 TI - H. pylori and reflux esophagitis in Turkish patients living in the Zaanstreek region in the Netherlands. AB - There are data on the prevalence is of reflux esophagitis in a population with a high prevalence of H. pylori infection. A cross-sectional study was done in a Turkish population in The Netherlands. A total of 1640 consecutive patients with reflux esophagitis were included. Sixty-one patients were of Turkish descent. Reflux esophagitis occurred significantly more often in ethnically Dutchpeople (overall 33% vs. 9.7%, P < 0.001). H. pylori was present in 60.6% of Turkish patients and in 18.5% of Dutch patients. All Turkish patients only suffered from mild esophagitis. It is concluded that the occurrence of reflux esophagitis is low in a population of Turkish patients with a high prevalence of H. pylori. PMID- 14561013 TI - Stimulation by capsaicin of duodenal HCO3(-) secretion via afferent neurons and vanilloid receptors in rats: comparison with acid-induced HCO3(-) response. AB - We compared the HCO3(-) secretory response to capsaicin and mucosal acidification in rat duodenums, especially the relation to vanilloid receptor type 1 (VR1). A proximal duodenal loop was perfused with saline, and the HCO3(-) secretion was measured at pH 7.0 using a pH-stat method and by adding 10 mM HCl. The secretion was stimulated by exposing the loop to capsaicin (0.03-0.3 mg/ml) or 10 mM HCl for 10 min. Indomethacin subcutaneously or ruthenium red intravenously, a nonspecific VR1 antagonist, was given 60 or 10 min, respectively, before exposure to capsaicin or acid, while L-NAME was given intravenously 3 hr before these treatments. Capsazepine, another VR1 antagonist, was coapplied to the loop for 10 min with capsaicin or acid. Luminal application of capsaicin increased the secretion of HCO3(-) in a dose-dependent manner; this effect was markedly attenuated by chemical ablation of capsaicin-sensitive afferent neurons (CSN) as well as pretreatment with ruthenium red or capsazepine, and significantly mitigated by indomethacin or L-NAME (in an L-arginine-sensitive manner). The HCO3(-) secretion was also stimulated by mucosal acidification, and this response was attenuated by both capsaicin pretreatment, indomethacin and L-NAME, but not ruthenium red or capsazepine. Mucosal application of capsaicin as well as acid increased the mucosal PGE2 content, and these effects were both significantly attenuated by indomethacin and L-NAME. These results suggest that both capsaicin and acid cause the CSN-dependent increase in duodenal HCO3(-) secretion mediated by NO and PG, yet the mode of their action differs in terms of the ruthenium red or capsazepine sensitivity. Although luminal H+ plays a modulatory role in duodenal HCO3(-) secretion, it is unlikely that the action results from the interaction of H+ with the ruthenium red- or capsazepine-sensitive site of VR1. PMID- 14561015 TI - Non-invasive positive pressure ventilation (NIPPV) in stable patients with chronic obstructive pulmonary disease (COPD). AB - While non-invasive positive pressure ventilation (NIPPV) has become an accepted management approach for patients with acute hypercapnia, it remains unclear whether it can also be beneficial in stable chronic obstructive pulmonary disease (COPD) patients with chronic respiratory failure. Randomised controlled trials (RCT) with a maximum duration of 3 months showed contradictory effects in blood gasses, dyspnoea, sleep efficiency and health-related quality of life. On the other hand, several uncontrolled trials did show positive results in patients with hypercapnia. Recently, an RCT compared the combination of NIPPV and long term oxygen treatment (LTOT) with LTOT alone for a period of 2 years in hypercapnic patients. After this period dyspnoea decreased and health-related quality of life improved in the NIPPV compared to the LTOT group. Reasons for the contradictory results in the different trials are probably patient selection, adequacy of ventilation, and length of ventilation. Therefore, at this moment there is no conclusive evidence that NIPPV should be provided routinely to stable patients with COPD. However, a selected group of patients might have clinical benefits from it. Patients who are clearly hypercapnic, who can tolerate an effective level of ventilatory support, and who get enough time to adjust to the ventilator might show clinical benefits even after 3 months. A trial with ventilatory support in this group of patients can be considered. PMID- 14561014 TI - Pulmonary alveolar microlithiasis: an overview of clinical and pathological features together with possible therapies. AB - Pulmonary alveolar microlithiasis is characterized by the presence in pulmonary alveolus of round shaped little bodies containing concentric calcareous lamellas. The incidence is similar in all continents, in both sexes and it is higher in age brackets between 20 and 50 years. The disease is prevalent among family units. Clinical reports may suggest the hypothesis that the disease may be hereditary. Pathogenetic hypotheses may indicate that a reduced lung mucociliary function leading to an excess of alveolar mucus may induce the formation of alveolar microliths by mucus condensation. Microliths may appear either confined in particular areas of the lung or widespread. Chemically, microliths consist of large amounts of calcium and phosphorus and, in reference to histology, they consist of calcareous concentric lamellas which are placed around an amorphous or granular central nucleus. The dissociation between definite X-ray pattern of lungs and relative poor clinical symptoms is the most common characteristics of the disease. However, a certain degree of dyspnea with a productive cough may occur together with a sporadic hemoptysis and thoracic pains. X-ray pattern of the lung reveals dissemination of radio-opaque nodules which may make lungs appear to be sprinkled with sand. The evolutive course of the disease leads to pulmonary insufficiency which is related to the increase of number of microliths in several areas of lungs. The inability to identify clear etiological and pathogenetic elements makes difficult therapeutic approach which is palliative such as the use of diphosphonate, steroids and therapeutic BAL. PMID- 14561016 TI - Cardioselective beta-blockers for chronic obstructive pulmonary disease: a meta analysis. AB - Beta-blocker therapy has a mortality benefit in patients with hypertension, heart failure and coronary artery disease, as well as during the perioperative period. These drugs have traditionally been considered contraindicated in patients with chronic obstructive pulmonary disease (COPD). The objective of this study was to assess the effect of cardioselective beta-blockers on respiratory function of patients with COPD. Comprehensive searches were performed of the EMBASE, MEDLINE and CINAHL databases from 1966 to May 2001, and identified articles and related reviews were scanned. Randomised, blinded, controlled trials that studied the effects of cardioselective beta-blockers on the forced expiratory volume in 1 s (FEV1) or symptoms in patients with COPD were included in the analysis. Interventions studied were the administration of beta-blocker, given either as a single dose or for longer duration, and the use of beta2-agonist given after the study drug. Outcomes measured were the change in FEV1 from baseline and the number of patients with respiratory symptoms. Eleven studies of single-dose treatment and 8 of continued treatment were included. Cardioselective beta blockers produced no significant change in FEV1 or respiratory symptoms compared to placebo, given as a single dose (-2.05% [95% CI, -6.05% to 1.96%]) or for longer duration (-2.55% [CI, -5.94% to 0.84]), and did not significantly affect the FEV1 treatment response to beta2-agonists. Subgroup analyses revealed no significant change in results for those participants with severe chronic airways obstruction or for those with a reversible obstructive component. In conclusion, cardioselective beta-blockers given to patients with COPD do not produce a significant reduction in airway function or increase the incidence of COPD exacerbations. Given their demonstrated benefit in conditions such as heart failure, coronary artery disease and hypertension, cardioselective beta-blockers should be considered for patients with COPD. PMID- 14561017 TI - Sleep apnoea and daytime function in the elderly--what is the impact of arousal frequency? AB - Arousals from sleep result in hyperventilation and hypocapnia that can lead to sleep apnoea. We have investigated whether sleep apnoea in the elderly is associated with more arousals compared with younger people. Additionally, the impact of arousals on daytime symptoms was noted. Four groups (n = 11) of elderly (> 65 years) and young (< 39 years) apnoeic (EA and YA), and age-matched non apnoeics (EN and YN) were studied. The arousal index (AI) and apnoea/hypopnoea index were determined from polysomnography. Sleepiness (Epworth Sleepiness Scale) and Quality of life (QoL, SF-36) were assessed. The mean (SD) AI was: EN 23.1 (7.6), EA 46.5 (8.8), YN 13.2 (6.6), YA 38.5 (12.1) events/h. AI was higher in the elderly (P = 0.002) and in apnoeics (P = 0.001); however, the increase in AI associated with sleep apnoea was not age dependent (P = 0.73). The influence of sleep apnoea on sleepiness was similar in both age groups. YA but not EA reported reduced physical functioning (P = 0.04), vitality (P = 0.007) and general health (P = 0.04) compared to non-apnoeics. We conclude that (1) the effect of sleep apnoea on arousal is no greater in the elderly compared to the young (2) despite similar levels of sleepiness, elderly apneoics perceive a reduced loss of QoL compared to younger patients. PMID- 14561018 TI - Mechanisms of basal and cytokine-induced uptake of glucose in normal human eosinophils: relation to apoptosis. AB - A link between glucose transport and apoptosis was suggested. We studied the mechanisms of glucose transport in human eosinophils by means of the uptake of the positron emitting analogue, 18Fluoro-2-Deoxyglucose (FDG) and apoptosis by means of flow cytometry. FDG uptake was inhibited by antibodies to GLUT1, 3 and 4 and by cytochalasin B. The anti-apoptotic principles IL-5, GM-CSF, IL-3 enhanced the uptake, whereas the apoptosis-inducing principles anti-CD95 (anti-Fas) and exposure to serum-coated Sephadex particles caused a reduction. Also TNF-alpha enhanced the uptake. Other cytokines such as IL-2, IL-4, IL-8, RANTES and MCP-3 had no effect on the glucose uptake. 2-Deoxyglucose, antibodies to GLUT4 and CD95 induced apoptosis. The basal FDG-uptake was unaffected by PKC inhibitors Ro-31 8220, Go-6983 and Go-6976, whereas the latter inhibited the IL-5-enhanced uptake possibly due to the inhibition of PKC(mu). Protein tyrosine kinase and PI-3 kinase inhibitors inhibited IL-5-enhanced FDG-uptake only. In contrast MEK inhibitors inhibited the basal uptake only. Inhibitors of p38 MAPkinase inhibited both basal and IL-5 enhanced uptake. We conclude that glucose uptake in eosinophils is governed by specific intracellular mechanisms involving mobilization of GLUTs, Ca2+ and the activation of the MAP kinase pathway and that the IL-5-enhanced uptake uniquely seems to involve PKC(mu) activity. Our results suggest a close link between apoptosis and glucose transport in human eosinophils. PMID- 14561019 TI - The perception of dyspnea after bronchoconstriction and bronchodilation in patients with asthma. AB - BACKGROUND: It is well documented that the perception of dyspnea (POD), subjectively reported by patients, is an important index used to guide treatment. The severity of dyspnea following methacholine-induced bronchoconstriction and added mechanical loads is increasing in popular. No formal attention has been addressed to the reduction in dyspnea following bronchodilators. STUDY OBJECTIVE: To investigate if the magnitude of dyspnea perceived by a subject is independent on the direction (e.g., bronchoconstriction or bronchodilation) of the change in airway resistance. METHODS: The POD was measured in 26 mild moderate asthmatic patients following bronchodilation, using beta2-agonists, and following bronchoconstriction, induced by methacholine challenge, to almost the same magnitude. RESULTS: The increase in forced expiratory volume in 1 s (FEV1), 30 min after the inhalation of beta2-agonist (mean +/- SEM 22.3 +/- 0.8%), was associated with a statistically significant decrease (P < 0.005) in the POD. The mean decrease in FEV1 following methacoline challenge, was 23 +/- 0.7% and was followed by a statistically significant increase (P < 0.005) in the POD. The magnitude of the decrease in the POD following albuterol was almost identical to the magnitude of the increase in the POD following methacholine. CONCLUSIONS: In stable mild-moderate asthmatic patients, the changes in the magnitude of dyspnea, perceived by a subject, is independent on the direction of the change in the FEV1. PMID- 14561020 TI - Assessment of patient performance of the HandiHaler compared with the metered dose inhaler four weeks after instruction. AB - The HandiHaler is a novel breath-actuated dry powder system designed for the delivery of tiotropium 18microg daily in the treatment of COPD. We compared patient ability to use the HandiHaler or metered dose inhaler (MDI) device correctly 4 weeks after receiving brief instructions and device demonstration. A single-blind study was conducted in COPD patients in two centers in Denmark. ALL patients (n = 151) received one placebo capsule via the HandiHaler daily and ipratropium (20 microg) two actuations via the MDI q.i.d. Mean FEV1 for all patients was 1.25 + 0.54 (46% predicted). Twelve instructions establishing proper device use were evaluated for the MDI and Handihaler. Error scores were analyzed by number of patients with less, equal or more errors when using HandiHaler compared to MDI in the total efficacy population (n = 139) and according to those who had not previously used an MDI for at least 12 months (MDI beginners) (n = 74) and those who had used an MDI (MDI experienced) (n = 65). Four weeks after device instruction, a higher proportion of patients in the total population (P < 0.01) had fewer errors with the HandiHaler (35.3%) compared to the MDI (15.1%). The number of errors was equal in 50% of patients. Similar findings were observed in the subgroup of patients who were MDI beginners (42% vs. 11%, P < 0.01) with non-significant trends in favor of the HandiHaler in those patients who were MDI experienced (29.7% vs. 18.9%, P = 0.096). Similar results in favor of HandiHaler were noted across different age and sex strata. The proportion of patients correctly using the device on the first of three attempts was 59.7% and 54.7% for the HandiHaler and MDI, respectively (P = 0.399). In summary, use of the HandiHaler can be easily taught with fewer errors compared to the MDI. Furthermore, patient performance using the HandiHaler was superior to that with an MDI despite prior MDI experience and more frequent usage. PMID- 14561021 TI - Sequential IV/PO moxifloxacin treatment of patients with severe community acquired pneumonia. AB - BACKGROUND: IV/PO moxifloxacin was evaluated in the treatment of hospitalized patients with severe community-acquired pneumonia (CAP). METHODS: Data were pooled from two prospective, randomized studies. In the multinational study, patients received 7-14 days IV/PO moxifloxacin 400 mg QD or IV/ PO amoxicillin clavulanate 1200/625 mg TID +/- IV/PO clarithromycin 500 mg BID. In the North American study, patients received 7-14 days IV/PO moxifloxacin 400 mg QD, IV/ PO alatrofloxacin/trovafloxacin 200 mg QD, or IV/PO levofloxacin 500 mg QD. The primary endpoint was clinical success at the test-to-cure visit. Severe CAP was defined according to the 1993 ATS criteria. RESULTS: In the clinically valid population, clinical success rates were 88% (167/190) for moxifloxacin- and 83% (155/186) for comparator-treated patients (95% CI = -1.9%, 12.2%). Corresponding clinical success rates for the microbiologically valid population were 87% (59/68) and 84% (54/64), respectively (95% CI = 8.6%, 15.0%). A switch from IV to PO therapy was made by day 5 of therapy for 73% of moxifloxacin- vs. 60% of comparator-treated patients (P < 0.01). Clinical success rates were similar in a retrospective analysis using the revised 2001 ATS definition of severe CAP. Mortality rates were 6% (15/241) and 10% (24/238) in the moxifloxacin and comparator treatment groups, respectively. The incidence of drug-related adverse events was similar in both treatment groups. CONCLUSION: Sequential IV/PO moxifloxacin 400 mg QD is as safe and effective as other fluoroquinolones and a beta-lactam/macrolide combination for treating hospitalized patients with severe CAP. PMID- 14561022 TI - Overuse of acid suppressive therapy in hospitalised patients with pulmonary diseases. AB - OBJECTIVES: Overuse of acid suppressive therapy (AST) has been reported in hospitalised patients, but the use in specific patient categories is unexplored. We assessed the use of and indication for AST and upper endoscopic investigations in hospitalised patients on a pulmonary ward compared with patients on other wards. METHODS: 301 patients were enrolled in the study. 162 were hospitalised on a pulmonary ward with a control group consisting of 139 from both a surgical and general internal medicine ward. Adequate indications for AST were those strongly supported by medical literature. RESULTS: Among the 301 patients enrolled, 132 (44%) used AST. 78 (59%) had no adequate indication for AST. On the pulmonary ward 79 (49%) patients used AST, compared to only 10 (20%) on the internal medicine ward (P < 0.05). On the pulmonary ward 68% of the patients had no adequate indication for AST, which was more common than inappropriate use of ASTon the control wards (P < 0.05). The most common inadequate indication for AST was peptic ulcer prophylaxis during corticoidsteroid therapy. CONCLUSION: In hospitalised patients a significant overuse of AST was observed, particularly among pulmonary patients. More adequate use of AST can contribute to substantial savings for the health-care system. PMID- 14561023 TI - Sleep breathing disorders in patients with idiopathic Parkinson's disease. AB - STUDY OBJECTIVES: to investigate the presence of sleep breathing disorders in patients with idiopathic Parkinson's disease (PD) and their correlation with the severity of the disease. PARTICIPANTS: Fifteen patients (mean age 63 +/- 4 years) with idiopathic PD (Group A) and 15 healthy matched controls (Group B) were studied. All patients were under treatment with L-Dopa/Carbidopa and classified according to the UPDRS motor scale: 8 had mild disease (UPDRS < 12), 6 moderate (UPDRS: 12-22) and 1 severe (UPDRS > 22). MEASUREMENTS AND RESULTS: All participants underwent full night polysomnography (PSG). The sleep-wake history was assessed. Spirometry, maximal respiratory pressures and arterial blood gases were also measured. Snoring was more common in Group A patients (73.3% vs. 33.3%, p = 0.002). Among the parameters studied apnea hypopnea index (AHI), mean O2 saturation, minimum O2 saturation, REM% sleep and Arousal Index (Arousal Index) were statistically different between the two groups. Furthermore, 9 PD patients fulfilled the criteria for obstructive sleep apnea-hypopnea syndrome (OSAHS) predominately mild, 1 for central sleep apnea hypopnea syndrome (CSAHS) and 5 were normal. In all patients a marked reduction in percentage REM sleep was observed. Among the patients with OSAHS 5 had mild PD, 3 moderate and 1 severe. The patient with CSAHS had moderate disease. Finally, 3 patients with mild and 2 with moderate PD had no evidence of sleep breathing disorders. Correlations between severity of disease and sleep parameters are provided. CONCLUSION: Our results suggest that sleep breathing disorders, predominantly obstructive, seem to be common in PD and those events correlate with the severity of the disease. PMID- 14561024 TI - Response to Ringbaek et al. PMID- 14561025 TI - Vibration analysis of angle-ply laminated composite plates with an embedded piezoceramic layer. AB - An optical full-field technique, called amplitude-fluctuation electronic speckle pattern interferometry (AF-ESPI), is used in this study to investigate the force induced transverse vibration of an angle-ply laminated composite embedded with a piezoceramic layer (piezolaminated plates). The piezolaminated plates are excited by applying time-harmonic voltages to the embedded piezoceramic layer. Because clear fringe patterns will appear only at resonant frequencies, both the resonant frequencies and mode shapes of the vibrating piezolaminated plates with five different fiber orientation angles are obtained by the proposed AF-ESPI method. A laser Doppler vibrometer (LDV) system that has the advantage of high resolution and broad dynamic range also is applied to measure the frequency response of piezolaminated plates. In addition to the two proposed optical techniques, numerical computations based on a commercial finite element package are presented for comparison with the experimental results. Three different numerical formulations are used to evaluate the vibration characteristics of piezolaminated plates. Good agreements of the measured data by the optical method and the numerical results predicted by the finite element method (FEM) demonstrate that the proposed methodology in this study is a powerful tool for the vibration analysis of piezolaminated plates. PMID- 14561026 TI - Piezoelectric micromotor based on the structure of serial bending arms. AB - This paper presents a new piezoelectric micromotor based on the structure of serial bending arms. Serial bending arms are composed of two piezoelectric bimorphs with one end fixed and the other end free, driven by two signals of a biased square wave with a phase difference of pi/2. The free end of a cantilever arm will move along an elliptic orbit so that the cantilever is used to drive a cylinder rotor. The rotor's end surface contacts the free end of the cantilever, resulting in the rotor's rotation. There are six serial bending arms anchored on the base. The driving mechanism of the micromotor is proposed and analyzed. A new micromotor prototype, 5 mm in diameter, has been fabricated and characterized. The maximum rotational speed reaches 325 rpm, and the output torque is about 36.5 microNm. PMID- 14561027 TI - A piezoelectric-sound-resonance cavity for hydrogen gas detection. AB - A new concept for hydrogen gas (H2) detection has been developed. This concept is based on a piezoelectric-sound-resonance cavity (PSRC). Detection uses sound resonance and the acoustic property differences of gases as a sensing mechanism in which there is a significant difference in the sound velocity and acoustic impedance between H2 and air. The PSRC sensor consists of two thin piezoelectric discs, separated by a small cavity. One disc excites a weak acoustic standing wave in the cavity, and the second senses the wave by monitoring the acoustic impedance characteristics of the cavity. Changes in H2 concentration result in a shift of the sound resonance state. Investigations of a PSRC prototype have demonstrated a sensitivity limit of <10 ppm, a fast response time approximately 1 second, and a signal (phase and voltage) in proportion to the change in H2 concentration (n) over the range of 10(-5) < n < 0.2. These performance characteristics are far superior to those of other methods. PMID- 14561028 TI - Thickness-shear vibrations of a quartz plate under time-dependent biasing deformations. AB - Incremental thickness-shear vibrations of a Y-cut quartz crystal plate under time harmonic biasing extensional deformations are studied using the two-dimensional equations for small fields superposed on finite biasing fields in an electroelastic plate. It is shown that the incremental thickness-shear vibrations are governed by the well-known Mathieu's equation with a time-dependent coefficient. Both free and electrically forced vibrations are studied. Approximate analytical solutions are obtained when the frequency of the biasing deformation is much lower than that of the incremental thickness-shear vibration. The incremental thickness-shear free vibration mode is shown to be both frequency and amplitude modulated, with the frequency modulation as a first-order effect and the amplitude modulation a second-order effect. The forced vibration solutions show that both the static and motional capacitances become time dependent due to the time-harmonic biasing deformations. PMID- 14561029 TI - Pulse-inversion-based fundamental imaging for contrast detection. AB - Pulse-inversion-based fundamental imaging was experimentally investigated for the enhancement of contrast detection. The pulse-inversion technique involves two firings with inverted waveforms. When the returning echoes from the two firings are summed, the residue signal is limited to even-order harmonics for tissue. However, when the returning echoes are from microbubbles, the fundamental signal is not completely cancelled because the reaction of the bubbles under compression is different from that under rarefaction. Thus, with the application of pulse inversion technique, the fundamental signal can be used to enhance the contrast to-tissue ratio. In this paper, B-mode, pulse-inversion-based fundamental images were constructed with various transmit waveforms. Motion artifacts also were studied. The results indicate that the contrast-to-tissue ratio was significantly enhanced compared to that obtained using either conventional, fundamental imaging or second-harmonic imaging. Longer transmit pulses resulted in a better signal-to noise ratio, but did not noticeably affect the nonlinear response of the bubbles. In addition, the optimal ratio of the magnitude of the positive pulse to that of the negative pulse was unity, in terms of avoiding the uncancelled, third-order response in the fundamental frequency range. It also was found that the pulse inversion fundamental technique is highly sensitive to tissue motion because the fundamental tissue signal is not cancelled when motion is present. PMID- 14561031 TI - Modeling and computer simulation of ultrasonic/sonic driller/corer (USDC). AB - Simulation and analytical models for the ultrasonic/sonic drill/corer (USDC) are described in this paper. The USDC was developed as a tool for in-situ rock sampling and analysis in support of the NASA planetary exploration program. The USDC uses a novel drive mechanism, which transfers ultrasonic vibrations of a piezoelectric actuator into larger oscillations of a free-flying mass (free mass). The free-mass impact on the drill bit creates a stress pulse at the drill tip/rock interface causing fracture in the rock. The main parts of the device (transducer, free-mass, bit, and rock) and the interactions between them were analyzed and numerically modeled to explore the drive mechanism. Each of these interactions is normally described by a time-dependent 2- or 3-D model involving slowly converging solutions, which makes the conventional approach unsuitable for USDC optimization studies. A simplified integrated model using tabulated data was developed to simulate the operation of the USDC on desktop PC and successfully predicted the characteristics of the device under a variety of conditions. The simulated results of the model and the experimental data used to verify the model are presented. PMID- 14561030 TI - Effects of acoustic heterogeneity in breast thermoacoustic tomography. AB - The effects of wavefront distortions induced by acoustic heterogeneities in breast thermoacoustic tomography (TAT) are studied. Amplitude distortions are shown to be insignificant for different scales of acoustic heterogeneities. For wavelength-scale, or smaller, heterogeneities, amplitude distortion of the wavefront is minor as a result of diffraction when the detectors are placed in the far field of the heterogeneities. For larger-scale heterogeneities at the parenchyma wall, by using a ray approach (geometric optics), we show that no refraction-induced multipath interference occurs and, consequently, that no severe amplitude distortion, such as is found in ultrasound tomography, exists. Next, we consider the effects of phase distortions (errors in time-of-flight) in our numerical studies. The numerical results on the spreads of point sources and boundaries caused by the phase distortions are in good agreement with the proposed formula. After that, we demonstrate that the blurring of images can be compensated for by using the distribution of acoustic velocity in the tissues in the reconstructions. The effects of the errors in the acoustical velocities on this compensation also are investigated. An approach to implement the compensation using only TAT data is proposed. Lastly, the differences in the effects of acoustic heterogeneity and the generation of speckles in breast TAT and breast ultrasound imaging are discussed. PMID- 14561033 TI - Single crystal PMN-0.33PT/epoxy 1-3 composites for ultrasonic transducer applications. AB - Lead magnesium niobate-lead titanate 0.67Pb (Mg(1/3)Nb(2/3))O3-0.33PbTiO3 (PMN 0.33PT, abbreviated as PMN-PT) single crystals were used to fabricate PMN PT/epoxy 1-3 composites with different volume fractions of PMN-PT ranging from 0.4 to 0.8. The electromechanical properties of the 1-3 composites were determined by the resonance technique. Theoretical modeling of the 1-3 composites matched quite well with the measured material properties. It was demonstrated that the thickness electromechanical coupling coefficients of the composites could reach as high as 0.8. A 2.4 MHz plane ultrasonic transducer was fabricated using a PMN-PT/epoxy 1-3 composite with 0.37 volume fraction of PMN-PT. It shows a -6 dB bandwidth of approximately 61% and an insertion loss of -14 dB. PMID- 14561032 TI - A high-frequency, 2-D array element using thermoelastic expansion in PDMS. AB - Optical generation of ultrasound is a promising alternative to piezoelectricity for high-frequency arrays. An array element is defined by the size and location of a laser beam focused on a suitable surface. Optical generation using the thermoelastic effect has traditionally suffered from low conversion efficiency. We previously demonstrated an increase in conversion efficiency of nearly 20 dB with an optical absorbing layer consisting of a mixture of polydimethylsiloxane (PDMS) and carbon black spin coated onto a glass microscope slide. Radiation pattern measurements with an 85 MHz spherically focused transducer indicated an array element size of 20 microm. These measurements lacked the spatial resolution required to reveal fine details in the radiated acoustic field. Here we report radiation pattern measurements with a 5-microm spatial sampling, showing that the radiated acoustic field is degraded by leaky Rayleigh waves launched from the PDMS/glass interface. We demonstrate that replacing the glass with a clear PDMS substrate eliminates the leaky Rayleigh waves, producing a broad and smooth radiation pattern suitable for a two-dimensional (2-D) phased array operating at frequencies greater than 50 MHz. PMID- 14561034 TI - A new regime for operating capacitive micromachined ultrasonic transducers. AB - We report on a new operation regime for capacitive micromachined ultrasonic transducers (cMUTs). Traditionally, cMUTs are operated at a bias voltage lower than the collapse voltage of their membranes. In the new proposed operation regime, first the cMUT is biased past the collapse voltage. Second, the bias voltage applied to the collapsed membrane is reduced without releasing the membrane. Third, the cMUT is excited with an ac signal at the bias point, keeping the total applied voltage between the collapse and snapback voltages. In this operation regime, the center of the membrane is always in contact with the substrate. Our finite element methods (FEM) calculations reveal that a cMUT operating in this new regime, between collapse and snapback voltages, possesses a coupling efficiency (k(T)2) higher than a cMUT operating in the conventional regime below its collapse voltage. This paper compares the simulation results of the coupling efficiencies of cMUTs operating in conventional and new operation regimes. PMID- 14561035 TI - Capacitive micromachined ultrasonic Lamb wave transducers using rectangular membranes. AB - This paper details the theory, fabrication, and characterization of a new Lamb wave device. Built using capacitive micromachined ultrasonic transducers (CMUTs), the structure described uses rectangular membranes to excite and receive Lamb waves on a silicon substrate. An equivalent circuit model for the transducer is proposed that produces results, which match well with those observed by experiment. During the derivation of this model, emphasis is placed on the resistance presented to the transducer membranes by the Lamb wave modes. Finite element analysis performed in this effort shows that the dominant propagating mode in the device is the lowest order antisymmetric flexural wave (A0). Furthermore, most of the power that couples into the Lamb wave is due to energy in the vibrating membrane that is transferred to the substrate through the supporting posts of the device. The manufacturing process of the structure, which relies solely on fundamental IC-fabrication techniques, is also discussed. The resulting device has an 18-microm-thick substrate that is almost entirely made up of crystalline silicon and operates at a frequency of 2.1 MHz. The characterization of this device includes S-parameter and laser vibrometer measurements as well as delay-line transmission data. The insertion loss, as determined by both S-parameter and delay-line transmission measurements, is 20 dB at 2.1 MHz. When configured as a delay-line oscillator, the device functions well as a sensor with sensitivity to changes in the mass loading of its substrate. PMID- 14561036 TI - Simple design rules for optimal design of dielectric temperature-compensated sapphire resonators. AB - It has been shown that the use of two dielectric crystals with opposite temperature coefficient of permittivity allows the realization of a resonator with a zero temperature coefficient of frequency. By using sapphire and rutile materials, which have low-loss tangents, some compensated resonators with very high Q-factors have been realized. In this work we develop rules that greatly simplify the design of a dielectric-compensated resonator. We show that the optimum design for compensation at a specific temperature may be determined by simply selecting the aspect ratio of the sapphire resonator. PMID- 14561037 TI - A study on phase-noise reduction method in phase-locked loop systems. AB - Experimental studies are carried out on phase noise and the correlation coefficient between the phase and average current noises of voltage-controlled oscillator in phased-locked loop (PLL) systems. The precise phase stabilization technique is discussed, and new methods to reduce the phase noise are described in PLL systems, using the correlation. PMID- 14561038 TI - Influence of a static magnetic field on the detected atomic velocity distribution in an optically pumped cesium beam frequency standard. AB - Irregularities of the microwave Ramsey patterns in an optically pumped cesium atomic beam frequency standards with a sharp angle incidence detecting laser beam have been diagnosed. They arise from a separated two-peak detected velocity distribution, which is caused by static magnetic fields. Experimentally measured results and theoretical analysis of this phenomenon are presented in this paper. PMID- 14561039 TI - Air-coupled ultrasonic tomographic imaging of high-temperature flames. AB - This paper illustrates the use of air-coupled ultrasonic tomography for the measurement of a high-temperature flame from a natural gas burner, using capacitive ultrasonic transducers in through transmission. This uses a transducer pair, which is scanned in two-dimensional sections at several angles to the jet axis. Travel-time data then is recorded along various paths in counter propagating directions. By processing the data obtained from propagation times, images have been formed of variations in temperature within the flame, using the tomographic reconstruction approach. PMID- 14561040 TI - Proton-exchanged wet etching of recessed-structure SAW filter. AB - An improved method for fabricating recessed-structure surface acoustic wave (SAW) filters is reported. The relation between proton-exchanged duration and etched depth are studied, as well as the relation between internal reflection, etched depth, and thickness of the aluminum electrode. Experimental results show that the devices have superior performance, and ideas for further improvement of the fabricated SAW device are discussed. PMID- 14561041 TI - Public health research among immigrant populations: still a long way to go. PMID- 14561042 TI - Are Russians different than other Europeans in their relation of risk factors to cardiovascular disease risk. PMID- 14561044 TI - Development of a semi-quantitative Food Frequency Questionnaire (FFQ) to assess dietary intake of multiethnic populations. AB - The Food Frequency Questionnaire (FFQ) is one of the most commonly used methods in epidemiological studies to assess long-term nutritional exposure. The purpose of this study is to develop a general FFQ for the Israeli population and present the procedures undertaken to select foods to be included in a questionnaire for a multiethnic population. Random population cluster sampling was done using voter registration lists of the Negev Jewish population, aged 35 years and over, which were divided into three groups based on where they were born, namely: Israel, Asia Africa and Europe-America. Participants were interviewed for their dietary intake using 24 hour recalls. Foods eaten by our subjects were aggregated into conceptually similar food groups and entered in stepwise regression models to predict variation in nutrient intake. Separate models were created for each origin group. It was found that the dietary intake was significantly lower for energy and vitamin E among European-American born subjects and significantly lower for calcium intake among Asian-African born subjects. Differences between ethnic groups were seen in the list of foods, which explained the between-person variability for energy and zinc. For most nutrients, fewer items were needed to explain the between-person variation in the group of people born in Israel. The final list of foods included 126 items and explained over 90% of the between person variability in selected nutrients for all three groups. The newly developed FFQ for the Negev population includes 126 items that are sufficient to rank the nutritional exposure of people over 35 years old from the three origin groups in Israel. PMID- 14561045 TI - Contraceptive practices of non-HIV-seropositive injecting drug users. AB - AIMS: To study the sociodemographic characteristics and sexual behaviours in contraception choices of injecting drug users (IDUs) and to compare the contraceptive practices of non-HIV-positive IDUs to those of the general population. DESIGN: Two surveys were used: a sample of IDUs attending 10 drug abuse treatment centres in the Paris region (IDU) and the Parisian subsample of the National French Survey of Sexual Behaviour (ACSF). MEASUREMENTS: Percentages of contraception practices were estimated separately for 81 IDU and 130 ACSF women, and for 175 IDU and 168 ACSF men, aged 25-34, not reporting prostitution or HIV seropositivity. FINDINGS: Most IDU (77%) and ACSF (84%) women, and IDU (73%) and ACSF (75%) men currently used a contraceptive method. Male condoms were more widely used by IDUs than by the general population (64 vs. 10% in women, 75 vs. 14% in men), for all subgroups of educational level, marital status, recent multipartnership status and sexual activity. CONCLUSIONS: Contraceptives are used as often by IDUs as by the general population aged 25-34. However, the methods employed differ, with higher condom use by IDUs, which suggest that IDUs take into account the risk of HIV contamination in their contraceptive practices. PMID- 14561046 TI - High cardiovascular mortality in Russia cannot be explained by the classical risk factors. The Arkhangelsk Study 2000. AB - Since the beginning of the 1990s the public health situation in Russia has been characterized by an extremely high mortality and a significant reduction in life expectancy. Cardiovascular diseases remained the major cause of death. Only a few large population studies were conducted in Russia during this period. A total of 1968 men and 1737 women aged 18-75 years participated in a health survey in Arkhangelsk, Russia, over the period 1999-2000. Investigation included assessment of classic cardiovascular risk factors (family history, smoking, blood pressure, and blood lipids) along with general health variables. The paper presents sex specific data on risk factors for coronary heart disease. Though the cardiovascular mortality is high in Russia, the calculated risk for coronary heart disease (the Framingham risk score and the Norwegian risk score) was lower in all age groups of men and women in Arkhangelsk compared with studies from the Western Europe and USA. Our data suggest that high cardiovascular mortality in Russia may be driven not only by the classic risk factors for coronary heart disease. PMID- 14561043 TI - Epidemiology of age-related maculopathy: a review. AB - Age-related maculopathy (ARM) is a degenerative disease of the retina and the leading cause of incurable blindness and visual impairment in industrialized countries. By definition, ARM is confined to the age-category above 50 years. The aetiology of ARM is still unknown, despite intensive research on many fronts. In this paper, we provide a review of the epidemiology of ARM. The most prominent findings were an exponential increase in frequency with age, a significant familial and genetic component, and a strong association with smoking. Other risk factors that were found less consistently were atherosclerosis, low intake of antioxidant nutrients, and cataract extraction. Future studies, both observational and experimental, will hopefully identify more risk factors that are amenable to prevention. PMID- 14561047 TI - Prevalence of stroke and transient ischaemic attack in the elderly population of an Italian rural community. AB - OBJECTIVES: To evaluate the prevalence of stroke and transient ischaemic attack in the elderly population, including the oldest residents, of a rural Italian community. METHODS: A door-to-door survey was performed between April and October 2001 in 2390 subjects aged 65 years and over. A symptom questionnaire validated by medical records and neurological examination was employed. RESULTS: Complete information was available for 2260 subjects. The overall prevalence of stroke was 8.2% in males and 5.1% in females. In the very elderly, this increased to 10.7% in males and 10% in females and decreased only in males aged 90 years or over. The overall prevalence of transient ischaemic attack was 7% in males and 4.9% in females. This, too, increased with age, reaching 10.2% in males and 7.4% in females and decreased only in subjects of both sexes aged 85 years or over. CONCLUSIONS: An actual estimate of the high prevalence of cerebrovascular disease in the elderly population, even in very old subjects, was thus achieved. Appropriate health care services consequently need to be planned. PMID- 14561048 TI - Cohort differences in obesity-related health indicators among 70-year olds with special reference to gender and education. AB - The aim of this study was to describe cohort differences in health indicators among four birth cohorts of 70-year old men and women from Goteborg, Sweden, born in 1901/2, 1905/6, 1911/12, and 1922. With special reference to gender, education, and obesity, it is hypothesized that changes in health among elderly men and women may not be occurring in a uniform manner. The variables studied were: systolic and diastolic blood pressures, triglycerides, cholesterol, height, weight, body mass index, waist-hip ratio, physical inactivity, current smoking, and alcohol consumption, plus selected prevalent diseases. Logistic and linear regression models were used to test for secular trends and effect modification by gender. Most trends in metabolic and lifestyle indicators varied in relation to gender as well as education. For instance, later-born male cohorts were more overweight than earlier-born groups while the later-born female cohorts had similar relative weights but a more centralized fat patterning. These cohort differences further varied by education where later-born men with less education and later-born women with higher education tended to be more overweight, compared to earlier-born cohorts. Finally, significant cohort differences in previously diagnosed myocardial infarction, stroke, and diabetes mellitus at age 70 were observed in men only. Interaction terms revealed that the gender difference was statistically significant only in the case of diabetes mellitus. In conclusion, secular trends in many obesity-related health indicators among 70-year old Swedish cohorts were dependent on both gender and socio-economic factors. PMID- 14561049 TI - Increased physical activity decreases periodontitis risk in men. AB - BACKGROUND: Increased physical activity improves insulin sensitivity and glucose metabolism, and may therefore affect incidence of periodontitis. METHODS: We studied the association of physical activity, walking and periodontitis in 39,461 male, US based, health professionals, 40-75 years old at baseline, more than half of whom were dentists, being followed up continuously since 1986. Participants were free of periodontitis, coronary heart disease and stroke at the start of follow-up. Physical activity and periodontitis were measured by validated questionnaires (expressed in metabolic equivalents--METs); the first report of professionally diagnosed periodontitis was considered a case. RESULTS: Periodontitis risk decreased by 3% for every 10-MET increase in average physical activity after adjustment for age, smoking, diabetes, BMI, alcohol consumption and total calories (RR = 0.97; 95% CI: 0.95-0.99). The inverse trend remained significant in the categorical analysis. Compared to men in the lowest quintile of physical activity, those in the highest quintile had a 13% lower risk of periodontitis (RR = 0.87; 95% CI: 0.76-1.01, p-value, test for trend = 0.02). In a sub-sample of men with radiographs (n = 137) the physically active had less average bone loss (beta = -0.29, p-value = 0.03) after multivariate adjustment compared to those inactive. CONCLUSIONS: In this large-scale prospective study, we found an inverse, linear association between sustained physical activity and periodontitis independent of known risk factors. The benefits of a physically active lifestyle may extend to periodontal health. PMID- 14561050 TI - Is asthma in 2-12 year-old children associated with physician-attended recurrent upper respiratory tract infections? AB - In a prevalence study, we evaluated whether recurrent physician-attended URTI episodes are more common in asthmatic children as compared to age- and gender matched controls. URTI proneness, defined as > or = 5 episodes of rhinitis/pharyngitis, sinusitis, laryngitis/tracheitis or otitis media in a 24 month period, was more common in asthmatics than controls (adjusted odds ratio 2.5, 95% confidence interval: 1.1-6.1). As a consequence, antibiotic prescriptions and referrals to hospitals occurred more frequently among asthmatics than controls. PMID- 14561051 TI - Epidemiology of genital herpes (HSV-2) among brothel based female sex workers in Bangladesh. AB - A prospective study was carried out to determine the prevalence of herpes simplex type 2 (HSV-2) in Bangladeshi female sex workers. Serum samples were collected from 463 female prostitutes and tested by ELISA using type specific antigens (gG1 and gG2). There were 405 (87.5%) samples seropositive for both HSV-1 and HSV-2, 24 (5.2%) samples were seropositive for only HSV-1, a further 33 (7.1%) samples were seropositive for HSV-2 only and one sample was found to be negative. Human immunodeficiency virus (HIV) testing was also performed and all samples were found to be negative. Surveillance studies are needed for development and application of control and preventative measures. The prostitute population in Bangladesh is particularly at risk of acquiring and transmitting HSV-2 and other sexually transmitted diseases (STDs). PMID- 14561052 TI - Distribution and prevalence of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks of canton Ticino (Switzerland). AB - Free-living Ixodes ricinus ticks were collected from 12 different sites of canton Ticino, south of the Alps (Switzerland). Each tick was examined for the presence of Borrelia burgdorferi sensu lato (sl), the etiologic agent of Lyme borreliosis using direct fluorescent antibody assay, and isolation of the bacteria. Borreliae were characterized by PCR followed by RFLP. The abundance and infection rates of I. ricinus ticks varied greatly between the areas. Two localities were found free of Borrelia. The prevalence of infected ticks ranged from 5 to 19%. Most ticks (96%) were found infected by < 50 spirochetes. Three B. burgdorferi sl species were successfully isolated: B. garinii dominated, followed by B. lusitaniae and B. valaisiana. Additionally, a mixed infection with B. garinii and B. valaisiana was observed. The distribution of the various Borrelia species in the different areas was heterogeneous. This is the first report of the presence of B. lusitaniae in I. ricinus in Switzerland. PMID- 14561053 TI - Acute brucellosis presenting with erythema nodosum. AB - Brucellosis is a common world-wide zoonotic disease. Cutaneous manifestations are not specific and affect 1-14% of patients with brucellosis. We describe two cases of young males presenting with skin lesions of erythema nodosum on the anterior surface of the legs. Histopathology of skin biopsy revealed septal panniculitis, but the positive cultures of blood or bone marrow for Brucella melitensis established the diagnosis of brucellosis. PMID- 14561054 TI - Risk of Haemophilus influenzae type b meningitis in Polish children varies directly with number of siblings: possible implications for vaccination strategies. AB - BACKGROUND: In the absence of an effective vaccine, Haemophilus influenzae type b (Hib) meningitis has frequently been found to be the most common bacterial meningitis among children less than or equal to 5 years of age. This study was conducted prior to the introduction of Hib vaccine in Poland, and is the first case-control study of Hib meningitis in this country. METHODS: A 1:3 matched case control study was nested within a population-based surveillance study of Hib meningitis in children less than or equal to 5 years old in Poland. Controls were matched on the case's age at onset of disease and immunization center. RESULTS: Having one or more siblings is a highly significant risk factor for Hib meningitis and, under the rare disease assumption, risk increases linearly with the increase in the number of siblings. The size of the living area of the home was not itself a significant risk factor for disease. Breast-feeding was not protective in contrast to previous studies. Childcare outside of the home was a significant risk factor for Hib meningitis especially among children greater than 16 months of age, whereas the effect of sibling number on Hib meningitis was much greater among the younger children. CONCLUSION: Risk of Hib meningitis approximately doubles for every unit increase in the number of siblings. Routine vaccination of Polish infants who have two or more siblings could potentially prevent half of the Hib meningitis cases. PMID- 14561055 TI - The changing profile of sickle cell disease in Italy. PMID- 14561056 TI - Moderators of the relation between popularity and depressive symptoms in children: processing strength and friendship value. AB - Children with low (n = 25) and high (n = 38) peer-rated popularity completed an emotional Stroop task, using negative social words, a self-report measure of friendship value relative to other domains of competence, and the Child Depression Inventory (CDI). Six months later, they completed the CDI again. In regression analyses, after controlling for prior CDI scores, social status interacted significantly with both Stroop and value measures (separate regressions). For unpopular children, both greater friendship valuing and greater negative social word Stroop interference predicted increases in depressive symptoms. In contrast, neither predictor was significant for popular children. In a third regression that included friendship value and Stroop interference as joint predictors of depression change, their effects remained significant and independent. We discuss these findings' implications for 3 models of depression; Champion and Power's social-cognitive theory of depression (L. A. Champion and M. J. Power, 1995), Pyszczynski and Greenberg's self-regulatory perseveration theory of depression (T. Pyszczynski & J. Greenberg, 1992), and Harter's model of global self-worth (S. Harter, 1985). PMID- 14561057 TI - Mothers' overreactive discipline and their encoding and appraisals of toddler behavior. AB - The relations of observed overreactive discipline with mothers' tendencies to notice negative, relative to positive, child behavior (preferential negative encoding), and mothers' negative appraisals of neutral and positive child behavior (negative appraisal bias), were examined in mothers of toddlers. The mothers rated both their own children's and unfamiliar children's behavior. Negative appraisal bias with respect to mothers' own (but not unfamiliar) children was related to mothers' overreactivity, independent of child misbehavior. Overreactivity was not related to mothers' preferential negative encoding either of their own or of unfamiliar children's behavior. However, in the case of mothers' own children, preferential negative encoding moderated the relation between negative appraisal bias and overreactive discipline, such that the negative appraisal bias-overreactivity relation was significant only in the context of high preferential negative encoding. PMID- 14561058 TI - The Infant-Toddler Social and Emotional Assessment (ITSEA): factor structure, reliability, and validity. AB - In this paper the refinement and psychometric properties of the Infant-Toddler Social and Emotional Assessment (ITSEA) are described. Results from a sociodemographically diverse birth cohort sample of 1,235 parents of children between the ages of 12 and 36 months are presented. Confirmatory factor analyses supported the hypothesized Internalizing, Externalizing, Regulatory, and Competence domains as well as the 17 individual scales that comprise the ITSEA. Findings for 3 additional indices useful in identifying significant psychopathology are presented. Subgroup analyses revealed structural invariance and expected mean level differences across both child sex and 6-month age bands. Child sex differences emerged for some problem and most competence scales, with boys rated as higher on Activity/Impulsivity and girls rated higher on Anxiety and most Competence scales. All competence scores increased across age groups. Problem behaviors showed a more mixed developmental pattern. Test-retest and interrater reliability were acceptable. Associations between the ITSEA and independent evaluator ratings and parental ratings of child behavior problems, temperament, and parental distress support the validity of the instrument. PMID- 14561059 TI - Parceling component effects of a multifaceted prevention program for disruptive elementary school children. AB - This study examined predictors and outcomes of attendance in two standard components of a multifaceted preventive intervention aimed at children with early onset disruptive behavior after 3 years of intervention. Mean rate of attendance in the Family Program, but not the Summer School Program, differed by level of child disruptiveness (grouping variable). Although predictors of attendance (SES, single-parent status, child IQ) did not differ across high- and low-disruptive groups, predictors of outcome were moderated by level of child disruptiveness for academic achievement and aggression outcomes, but not for social competence. Higher attendance in the Summer Program was associated with higher child social competence at Year 3 for all children. For academic achievement, higher attendance in the Summer Program was associated with higher scores for mild/moderately disruptive children and lower scores for highly disruptive children in Year 3. Higher attendance in the Family Program was associated with lower aggression scores for mild/moderately disruptive children. Findings highlight the importance of matching intervention components to the assessed or expressed needs of client subgroups. PMID- 14561060 TI - Convergent and discriminant validity of trait and source effects in ADHD inattention and hyperactivity/impulsivity measures across a 3-month interval. AB - Confirmatory factor analysis was used to model a multitrait (ADHD-inattention and hyperactivity/ impulsivity) by multisource (teachers and parents) design across a 3-month interval in a sample of 360 Australian elementary school children. The purpose was to evaluate the convergent and discriminant validity of the ADHD inattention (IN) and hyperactivity/impulsivity (H/I) measures. Although similar traits and similar sources showed stronger correlations across time than dissimilar traits and dissimilar sources, the amount of source variance in the ADHD-IN and ADHD-H/I measures was substantial and consistent across the interval (M = 59%; range = 35-84%). This large amount of source variance raises the possibility that the correlations of the IN and H/I rating scales with other constructs (e.g., social competence, conduct problems) represent mostly source rather than trait effects. Multitrait by multisource analyses provide a means to answer this question and further advance understanding of ADHD. PMID- 14561061 TI - Relations between Continuous Performance Test performance measures and ADHD behaviors. AB - The Conners' Continuous Performance Test (CPT) is a neuropsychological task that has repeatedly been shown to differentiate ADHD from normal groups. Several variables may be derived from the Conners' CPT including errors of omission and commission, mean hit reaction time(RT), mean hit RT standard error, d', and beta. What each CPT parameter actually assesses has largely been based upon clinical assumptions and the face validity of each measure (e.g., omission errors measure inattention, commission errors measure impulsivity). This study attempts to examine relations between various CPT variables and phenotypic behaviors so as to better understand the various CPT variables. An epidemiological sample of 817 children was administered the Conners' CPT. Diagnostic interviews were conducted with parents to determine ADHD symptom profiles for all children. Children diagnosed with ADHD had more variable RTs, made more errors of commission and omission, and demonstrated poorer perceptual sensitivity than nondiagnosed children. Regarding specific symptoms, generalized estimating equations (GEE) and ANCOVAs were conducted to determine specific relationships between the 18 DSM-IV ADHD symptoms and 6 CPT parameters. CPT performance measures demonstrated significant relationships to ADHD symptoms but did not demonstrate symptom domain specificity according to a priori assumptions. Overall performance on the two signal detection measures, d' and beta, was highly related to all ADHD symptoms across symptom domains. Further, increased variability in RTs over time was related to most ADHD symptoms. Finally, it appears that at least 1 CPT variable, mean hit RT, is minimally related to ADHD symptoms as a whole, but does demonstrate some specificity in its link with symptoms of hyperactivity. PMID- 14561062 TI - Continuous performance test performance in a normative epidemiological sample. AB - A 14-min continuous performance test (CPT) requiring a high rate of responding was administered to a probability-weighted random sample of 816 9-17-year-old children drawn from a population of 17,117 children in an ongoing epidemiological and longitudinal study in Western North Carolina. Systematic main effects of improved performance with older age were found in this age range for all variables, including reaction time (RT), RT standard error, errors of omission, errors of commission, and signal detection parameters (d' and beta). Significant gender main effects included more impulsive errors, less variability, and faster RT by males, with no interactions between age and gender. There were no main effects of ethnicity or interactions of ethnicity with age and/or gender. Large main effects of interstimulus interval (ISI; 1, 2, or 4-s intervals) and time block were present for most CPT performance measures. The normative data from the CPT should provide a useful framework for interpreting similar data in future studies of child and adolescent psychopathology. PMID- 14561063 TI - Insect peptide hormones: a selective review of their physiology and potential application for pest control. AB - Our knowledge on primary structure, synthesis, release, receptor binding, structure-activity relationships, mode of action and degradation of, mainly, neuropeptides from insects has increased dramatically during the last 10 years or so. Here, five case studies are presented, which deal selectively with effects on: reproduction (trypsin modulating oostatic factor in mosquito); energy metabolism, locomotion and the immune system (adipokinetic hormones); water and ion balance, and feeding behaviour (diuretic hormones, kinins, sulfakinins); sex attraction (pheromone biosynthesis activating neuropeptide); and growth and development, and muscle activity (allatostatins). The literature is reviewed in the context of how the knowledge on neuropeptides has been and can be used for the design of novel, safe and selective compounds to control pest insects in the foreseeable future. PMID- 14561064 TI - Capacity of model biobeds to retain and degrade mecoprop and isoproturon. AB - Biobeds are used to increase the adsorption and degradation of pesticide spillage on sites used for mixing and loading and for cleaning of sprayers. The adsorption and the rate of degradation of 14C-labelled isoproturon and mecoprop (MCPP) at concentrations from 0.0005 to 25 000 mgkg(-1) were determined in biobed soil. Further leaching of the two herbicides was determined in a model biobed with a surface area of 2 m2. The biobed material showed enhanced ability to adsorb the two herbicides. Kd was 5.2 litre kg(-1) for isoproturon and 1.6 litre kg(-1) for MCPP in biobed material, which is higher than in natural soil. In different experiments with natural soil, Kd ranges from 0.07 to 0.6 litrekg(-1) for MCPP and from 1.5 to 4.6 litre kg(-1) for isoproturon in soils with varying organic carbon content. Degradation of MCPP was rapid at concentrations from 0.0005 to 500 mg kg(-1), delayed at 5000 mg kg(-1), and very slow at 25 000 mg kg(-1). For isoproturon, the relative degradation was most rapid at the lowest concentration and decreasing with increasing concentrations. After 120 days, between 55% and 8% 14C was evolved as 14CO2 at concentrations between 0.0005 and 25 000 mg kg(-1). The rate of evolution of 14CO2 indicated that degradation rates at low concentrations were of first-order and at higher concentrations of zero-order. Leaching of MCPP and isoproturon was determined in a newly established model biobed during a 2-year period. About 13% of applied MCPP and 1.4% of applied isoproturon leached out during the winter following the first autumn application (worst-case scenario). Leaching was completely prevented when the biobed had a well-developed grass cover and was covered during the winter. PMID- 14561065 TI - Effects of fludioxonil and pyrimethanil, two fungicides used against Botrytis cinerea, on carbohydrate physiology in Vitis vinifera L. AB - In Vitis vinifera L, photosynthesis and photosynthate partitioning are affected in the presence of fludioxonil and pyrimethanil, two fungicides commonly used in vineyards against Botrytis cinerea Pers. However, the effects were found to be different according to the model studied: plantlets (cv Chardonnay) grown in vitro, fruiting cuttings (cv Chardonnay) and plants grown in vineyards (cvs Chardonnay, Pinot noir and Pinot Meunier). In the plantlets grown in vitro, both fungicides decreased gas exchanges, photosynthetic pigment and starch concentrations in the leaves, whereas soluble carbohydrates transiently accumulated, suggesting that plantlets mobilised starch in response to photosynthesis inhibition caused by fungicides. In the fruiting cuttings, the fungicides did not affect photosynthesis, although fludioxonil caused starch decrease in parallel with sucrose accumulation, suggesting that the fungicide effects were of lower intensity than in vitro. Conversely, in vineyard, the two fungicides stimulated photosynthesis and increased pigment concentrations in the three vine cultivars tested. In the meantime, glucose, fructose and starch levels of the leaves declined after fungicide exposure, whereas sucrose accumulated, indicating that sucrose synthesis increased in the leaves following the fungicide treatment. Among the three varieties, Chardonnay was the most sensitive to the fungicides as revealed by the intensity of the responses and the longer period for recovery. In vineyard, the results suggested that the two fungicides, in addition to inhibiting B cinerea development, had a beneficial effect on vine physiology through the stimulation of leaf carbon nutrition, which may further enable the plant to rapidly make use of its defence reactions. PMID- 14561066 TI - Insecticidal activity and nicotinic acetylcholine receptor binding of dinotefuran and its analogues in the housefly, Musca domestica. AB - The insecticidal activity of dinotefuran and 23 related compounds against the housefly, Musca domestica (L) was measured by injection with metabolic inhibitors. Dinotefuran was less active than imidacloprid and clothianidin by a factor of 10 in molar concentrations. Their binding activities to the fly-head membrane preparation were measured by using [125I]alpha-bungarotoxin ([125I]alpha BGTX) and [3H]imidacloprid ([3H]IMI) as radioligands. The activity of some selected compounds measured with [3H]IMI was 10(4)-fold higher than that measured with [125I]alpha-BGTX. With [3H]IMI as a radioligand, dinotefuran was 13-fold less active than imidacloprid. The inhibitory effect of dinotefuran on the binding of [3H]IMI to the membrane preparation was in a competitive manner. Quantitative analysis of the insecticidal activity of the test compounds with the binding activity measured with [3H]IMI showed that the higher the binding activity, the higher was the insecticidal activity. PMID- 14561067 TI - Predictability of combined effects of eight chloroacetanilide herbicides on algal reproduction. AB - Chloroacetanilides are pre-emergence herbicides for the control of annual grasses and broadleaf weeds. As a result of their extensive use, residues are often found in surface waters. Observed simultaneous occurrence of different chloroacetanilide herbicides gives reason for concern about potential combination effects on aquatic non-target organisms. This study aimed to clarify whether joint effects of various chloroacetanilide herbicides may be predictable from knowledge of concentration-response relationships of single substances. Whether the chloroacetanilides all share the same mode of action is unclear. Therefore we investigated the predictive value of two alternative concepts for the prediction of combined effects: concentration addition, which assumes a similar mode of action, and independent action, which is based on the idea of a dissimilar mode of action of the mixture components. Eight chloroacetanilides (acetochlor, alachlor, butachlor, dimethachlor, metazachlor, metolachlor, pretilachlor and propachlor) were experimentally tested for their individual as well as for their combined effects in mixtures on the reproduction of the green alga Scenedesmus vacuolatus. Individual chloroacetanilides impaired algal reproduction, with EC50 values ranging from 3 to 232 microg litre(-1). The differences in EC50 values were strongly correlated with the lipophilicities of the compounds. Effects of chloroacetanilide mixtures were considerably higher than those of the individual components: a complete inhibition of algal reproduction was observed when every mixture component was present in a concentration that would cause only 5% effect if applied singly. However, the combined effects proved to be predictable by using the concept of concentration addition. The alternative concept, independent action, distinctly underestimated the mixture toxicity. These findings (1) indicate a similar mechanism of action of chloroacetanilides in algae and (2) reinforce the view that concentration addition is a reasonable assumption for the predictive hazard assessment of groups of similarly acting herbicides. PMID- 14561068 TI - Inhibitors of 3-hydroxy-3-methylglutaryl-CoA reductase lower fecundity in the German cockroach: correlation between the effects on fecundity in vivo with the inhibition of enzymatic activity in embryo cells. AB - The enzyme 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase is crucial to insect development and reproduction, as revealed by the sterilising properties of some specific inhibitors of it. In the present paper, we study the sterilising effects of a number of HMG-CoA reductase inhibitors on the German cockroach, Blattella germanica (L). The inhibitors tested were naringenin, lovastatin, mevastatin, simvastatin, atorvastatin and fluvastatin. The first two compounds were ineffective or scarcely effective as HMG-CoA reductase inhibitors. The most active compounds in vivo were fluvastatin and atorvastatin, followed by simvastatin and mevastatin. They were equally ranked when tested as HMG-CoA reductase inhibitors in the B germanica embryonic derived cell line UM-BGE-1. This suggests that this cell line may be an appropriate tool for testing HMG-CoA reductase inhibitors and so to predict their properties as insect sterilising agents with insecticide potential. PMID- 14561069 TI - Rapid mineralisation of the herbicide isoproturon in soil from a previously treated Danish agricultural field. AB - Mineralisation of the phenylurea herbicide isoproturon (3-(4-isopropylphenyl)-1,1 dimethylurea) and two of its known metabolites, 3-(4-isopropylphenyl)-1 methylurea (monodesmethyl-isoproturon) and 4-isopropylaniline, was studied in Danish agricultural soils with or without previous exposure to isoproturon. A potential for rapid mineralisation of isoproturon and the two metabolites was present in soils sampled from three plots within an agricultural field previously treated regularly with the herbicide, with 34-45%, 51-58% and 33-36% of the added [phenyl-U-14C]isoproturon, [phenyl-U-14C]monodesmethyl-isoproturon and [phenyl-U 14C]4-isopropylaniline metabolised to [14C]carbon dioxide within 30 days at 20 degrees C. In contrast, such extensive mineralisation of these three compounds was not observed within this period in soils sampled from two other agricultural fields without previous treatment with isoproturon. The mineralisation patterns indicated growth-linked metabolism of the three compounds in the previously exposed soils, and doubling times for [14C]carbon dioxide production ranged from 1.6 to 3.2, 1.0 to 2.1 and 1.3 to 1.7 days for isoproturon, monodesmethyl isoproturon and 4-isopropylaniline, respectively. The ability to mineralise [phenyl-U-14C]isoproturon to [14C]carbon dioxide was successfully sub-cultured to a fresh mineral medium which provided isoproturon as sole source of carbon and nitrogen. One of the soils sampled from an agricultural field not previously treated with isoproturon showed accelerated mineralisation of [phenyl-U-14C]4 isopropylaniline toward the end of the experiment, with a doubling time for [14C]carbon dioxide production of 7.4days. This study indicates that the occurrence of rapid mineralisation of the phenyl ring of isoproturon to carbon dioxide is related to previous exposure to the herbicide, which suggests that microbial adaptation upon repeated isoproturon use may occur within agricultural fields. PMID- 14561070 TI - A degree-day concept for estimating degradation time under field conditions. AB - Development of degradation rate information for crop protection products in soil is typically a time consuming and costly process. Therefore, development of methodology that would allow extrapolation of limited degradation information to wider field conditions in a predictive fashion would be desirable. This study presents an approach that uses a modified heat unit model to estimate organic compound degradation time using limited datasets. The model requires daily soil or air temperature and volumetric water content. Predictions based on the model for three different compounds in five different field studies were within 80% of measured values based on soil analysis. Results from this research indicate that the model was able to predict compound half-life values across all sites with r2 = 0.99. PMID- 14561071 TI - Triazine resistance in Amaranthus tuberculatus (Moq) Sauer that is not site-of action mediated. AB - While surveying Illinois Amaranthus tuberculatus (Moq) Sauer (tall waterhemp) half-sib populations for herbicide response variability, several were observed to segregate for resistance to atrazine. Studies were conducted on greenhouse-grown A tuberculatus plants to compare atrazine responses among populations that were segregating for resistance (SegR), uniformly sensitive (UniS) or uniformly resistant (UniR). In chlorophyll fluorescence assays, leaves of plants from the SegR and UniS populations displayed changes in fluorescence after treatment with atrazine, indicating that atrazine was inhibiting electron transport of photosystem II in chloroplasts. Sequencing of a fragment of psbA, which encodes the D1 protein, revealed that the SegR population did not contain the amino acid substitution that is typically found in triazine-resistant plants. Whole-plant herbicide dose-response experiments revealed that, relative to the UniS population, atrazine resistances in the UniR and SegR populations were > 770-fold and 16-fold, respectively. The SegR population was also resistant to cyanazine (59-fold), but not to metribuzin, linuron or pyridate. Triazine resistance in the SegR population was shown to be a nuclear inherited trait, unlike maternal inheritance of site-of-action mediated triazine resistance found in the UniR population. Taken collectively, these findings confirm the existence of two distinct triazine resistance mechanisms in A tuberculatus. PMID- 14561072 TI - Cloning and expression analysis of the ATP-binding cassette transporter gene MFABC1 and the alternative oxidase gene MfAOX1 from Monilinia fructicola. AB - Brown rot, caused by Moniliniafructicola (G Wint) Honey, is a serious disease of peach in all commercial peach production areas in the USA, including South Carolina where it has been primarily controlled by pre-harvest application of 14 alpha demethylation (DMI) fungicides for more than 15 years. Recently, the Qo fungicide azoxystrobin was registered for brown rot control and is currently being investigated for its potential as a DMI fungicide rotation partner because of its different mode of action. In an effort to investigate molecular mechanisms of DMI and Qo fungicide resistance in M fructicola, the ABC transporter gene MfABC1 and the alternative oxidase gene MfAOX1 were cloned to study their potential role in conferring fungicide resistance. The MfABC1 gene was 4380 bp in length and contained one intron of 71 bp. The gene revealed high amino acid homologies with atrB from Aspergillus nidulans (Eidam) Winter, an ABC transporter conferring resistance to many fungicides, including DMI fungicides. MfABC1 gene expression was induced after myclobutanil and propiconazole treatment in isolates with low sensitivity to the same fungicides, and in an isolate with high sensitivity to propiconazole. The results suggest that the MfABC1 gene may be a DMI fungicide resistance determinant in M fructicola. The alternative oxidase gene MfAOX1 from M fructicola was cloned and gene expression was analyzed. The MfAOX1 gene was 1077 bp in length and contained two introns of 54 and 67 bp. The amino acid sequence was 63.8, 63.8 and 57.7% identical to alternative oxidases from Venturia inaequalis (Cooke) Winter, Aspergillus niger van Teighem and A nidulans, respectively. MfAOX1 expression in some but not all M fructicola isolates was induced in mycelia treated with azoxystrobin. Azoxystrobin at 2 microg ml(-1) significantly induced MfAOX1 expression in isolates with low MfAOX1 constitutive expression levels. PMID- 14561073 TI - Biotypic and pest status differences between Hungarian and South African populations of Russian wheat aphid, Diuraphis noxia (Kurdjumov) (Homoptera: Aphididae). AB - Russian wheat aphid, Diuraphis noxia (Kurdjumov) is a severe pest of cereals in South Africa and in the USA. In order to reduce D noxia damage, intensive resistance breeding programs have been undertaken, resulting in D noxia-resistant cultivars that are now widely used in South Africa and in the USA. However, there appear to be differences in the ability of different populations of D noxia to damage these resistant cereal cultivars. To determine whether different biotypes of D noxia are present, damage to eight wheat cultivars was compared when they were exposed to either Hungarian or South African aphid strains. It appeared that the Hungarian CVS MV Magdalena, MV Magvas, and MV 17 were susceptible to D noxia from both Hungary and South Africa. The susceptible South African CV Betta was also severely damaged regardless of the country of origin of the aphids. None of the cultivars resistant in South Africa (Caledon, SST 333, SST 972 and Halt) were, however, resistant to Hungarian populations of D noxia. These cultivars, which carry resistance genes originating from the breeding lines PI 262660, PI 137739 and PI 372129, were severely damaged by the Hungarian D noxia. Apart from the highly resistant CV Halt, the resistant cultivars used in this study were developed in South Africa, with the biotype present there. Damage to all cultivars tested was significantly more severe in response to Hungarian than to South African D noxia, indicating that a more damaging aphid biotype occurs in Hungary. However, D noxia has not yet become a pest of wheat in Hungary, possibly due to a difference in cultural practices. PMID- 14561074 TI - Effect of rotenoids from the seeds of Millettia dura on larvae of Aedes aegypti. AB - A crude chloroform extract of seeds of Millettia dura Dunn (Leguminosae) showed high activity (LC50 = 3.5 microg ml(-1) at 24 h) against second-instar larvae of the mosquito, Aedes aegypti L (Diptera: Culicidae). The rotenoids, deguelin and tephrosin, isolated from the seeds of this plant also showed potent activities, with LC50 values of 1.6 and 1.4 microg ml(-1) at 24 h, respectively. The related rotenoids millettone and millettosin were inactive at 20 microg ml(-1). Saturation at the B/C ring junction and the presence of methoxy groups at C-2 and/or C-3 in deguelin and tephrosin appear to be important for the observed larvicidal activity. PMID- 14561075 TI - The potential removal of imidacloprid from water by heat-treated kerolites. AB - The adsorption of imidacloprid [1-(6-chloro-3-pyridinylmethyl)-N nitroimidazolidin-2-ylideneamine] on heat-treated kerolite samples at 110 degrees C (K-110), 200 degrees C (K-200), 400 degrees C (K-400) and 600 degrees C (K-600) from pure water solution at 25 degrees C has been studied. The evolution of the surface properties of the kerolite samples, such as specific surface area and porosity, after heat treatment were analysed. The clays were characterised by using FTIR spectroscopy, X-ray diffraction, thermogravimetric and differential thermogravimetric analysis, surface analysis and Hg porosimetry. The experimental adsorption data points were fitted to the Freundlich equation in order to calculate the adsorption capacities (Kf) of the samples; Kf values ranged from 242 mg kg(-1) for the K-110 sample to 1005 mg kg(-1) for the K-600 sample. The values obtained for the removal efficiency (R) ranged from 62.8% for K-110 to 87.2% for K-600. The adsorption experiments showed that the stronger the heat treatment, the more effective was the adsorption of imidacloprid from pure water. This work shows the potential use of heat-activated kerolite for the removal of imidacloprid from environmental waters and drinking water resources. PMID- 14561076 TI - Veterinary medicines in the environment. AB - The impact of veterinary medicines on the environment will depend on a number of factors including physicochemical properties, amount used and method of administration, treatment type and dose, animal husbandry practices, manure storage and handling practices, metabolism within the animal, and degradation rates in manure and slurry. Once released to the environment, other factors such as soil type, climate, and ecotoxicity also determine the environmental impact of the compound. The importance of individual routes into the environment for different types of veterinary medicines varies according to the type of treatment and livestock category. Treatments used in aquaculture have a high potential to reach the aquatic environment. The main routes of entry to the terrestrial environment are from the use of veterinary medicines in intensively reared livestock, via the application of slurry and manure to land, and by the use of veterinary medicines in pasture-reared animals where pharmaceutical residues are excreted directly into the environment. Veterinary medicines applied to land via spreading of slurry may also enter the aquatic environment indirectly via surface runoff or leaching to groundwater. It is likely that topical treatments have greater potential to be released to the environment than treatments administered orally or by injection. Inputs from the manufacturing process, companion animal treatments, and disposal are likely to be minimal in comparison. Monitoring studies demonstrate that veterinary medicines do enter the environment, with sheep dip chemicals, antibiotics, sealice treatments, and anthelmintics being measured in soils, groundwater, surface waters, sediment, or biota. Maximum concentrations vary across chemical classes, with very high concentrations being reported for the sheep dip chemicals. The degree to which veterinary medicines may adsorb to particulates varies widely. Partition coefficients (K(d)) range from low (0.61 L kg(-1)) to high (6000 L kg(-1)). The variation in partitioning for many of the compounds in different soils was significant (up to a factor of 30), but these differences could be not be explained by normalization to the organic carbon content of the soils. Thus, to arrive at a realistic assessment of the availability of veterinary medicines for transport through the soil and uptake into soil organisms, the K(oc) (which is used in many of the exposure models) may not be an appropriate measure. Transport of particle-associated substances from soil to surface waters has also been demonstrated. Veterinary medicines can persist in soils for days to years, and half-lives are influenced by a range of factors including temperature, pH, and the presence of manure. The persistence of major groups of veterinary medicines in soil, manure, slurry, and water varies across and within classes. Ecotoxicity data were available for a wide range of veterinary medicines. The acute and chronic effects of avermectins and sheep dip chemicals on aquatic organisms are well documented, and these substances are known to be toxic to many organisms at low concentrations (ng L( 1) to microg L(-1)). Concerns have also been raised about the possibility of indirect effects of these substances on predatory species (e.g., birds and bats). Data for other groups indicate that toxicity values are generally in the mg L(-1) range. For the antibiotics, toxicity is greater for certain species of algae and marine bacteria. Generally, toxicity values for antibacterial agents were significantly higher than reported environmental concentrations. However, because of a lack of appropriate toxicity data, it is difficult to assess the environmental significance of these observations with regard to subtle long-term effects. PMID- 14561077 TI - Health effects of Acanthamoeba spp. and its potential for waterborne transmission. AB - Risk from Acanthamoeba keratitis is complex, depending upon the virulence of the particular strain, exposure, trauma, or other stress to the eye, and host immune response. Bacterial endosymbionts may also play a factor in the pathogenicity of Acanthamoeba. Which factor(s) may be the most important is not clear. The ability of the host to produce IgA antibodies in tears may be a significant factor. The immune response of the host is a significant risk factor for GAE infection. If so, then a certain subpopulation with an inability to produce IgA in the tears may be at greatest risk. There was no sufficient data on the occurrence or types of Acanthamoeba in tapwater in the U.S. Published work on amoebal presence in tapwater does not provide information on the type of treatment the water received or the level of residual chlorine. Assessment of the pathogenicity by cell culture and molecular methods of Acanthamoeba in tapwater would also be useful in the risk assessment process for drinking water. The possibility that Acanthamoeba spp. might serve as vectors for bacterial infections from water sources also should be explored. The bacterial endosymbionts include an interesting array of pathogens such as Vibrio cholerae and Legionella pneumophila, both of which are well recognized waterborne/water-based pathogens. Work is needed to determine if control of Acanthamoeba spp. is needed to control water-based pathogens in water supplies. PMID- 14561078 TI - Arsenic hazards to humans, plants, and animals from gold mining. AB - Arsenic sources to the biosphere associated with gold mining include waste soil and rocks, residual water from ore concentrations, roasting of some types of gold containing ores to remove sulfur and sulfur oxides, and bacterially enhanced leaching. Arsenic concentrations near gold mining operations are elevated in abiotic materials and biota: maximum total arsenic concentrations measured were 560 microg/L in surface waters, 5.16 mg/L in sediment pore waters, 5.6 mg/kg DW in bird liver, 27 mg/kg DW in terrestrial grasses, 50 mg/kg DW in soils, 79 mg/kg DW in aquatic plants, 103 mg/kg DW in bird diets, 225 mg/kg DW in soft parts of bivalve molluscs, 324 mg/L in mine drainage waters, 625 mg/kg DW in aquatic insects, 7,700 mg/kg DW in sediments, and 21,000 mg/ kg DW in tailings. Single oral doses of arsenicals that were fatal to 50% of tested species ranged from 17 to 48 mg/kg BW in birds and from 2.5 to 33 mg/kg BW in mammals. Susceptible species of mammals were adversely affected at chronic doses of 1-10 mg As/kg BW or 50 mg As/kg diet. Sensitive aquatic species were damaged at water concentrations of 19-48 microg As/L, 120 mg As/kg diet, or tissue residues (in the case of freshwater fish) > 1.3 mg/kg fresh weight. Adverse effects to crops and vegetation were recorded at 3-28 mg of water-soluble As/L (equivalent to about 25-85 mg total As/kg soil) and at atmospheric concentrations > 3.9 microg As/m3. Gold miners had a number of arsenic-associated health problems, including excess mortality from cancer of the lung, stomach, and respiratory tract. Miners and schoolchildren in the vicinity of gold mining activities had elevated urine arsenic of 25.7 microg/L (range, 2.2-106.0 microg/L). Of the total population at this location, 20% showed elevated urine arsenic concentrations associated with future adverse health effects; arsenic-contaminated drinking water is the probable causative factor of elevated arsenic in their urine. Proposed arsenic criteria to protect human health and natural resources are listed and discussed. Many of these proposed criteria do not adequately protect sensitive species. PMID- 14561079 TI - The winding road to the discovery of the SR/CR mice. PMID- 14561080 TI - Human beta-secretase (BACE) and BACE inhibitors. PMID- 14561081 TI - Thienyl and phenyl alpha-halomethyl ketones: new inhibitors of glycogen synthase kinase (GSK-3beta) from a library of compound searching. AB - Glycogen synthase kinase (GSK-3beta) plays a crucial role in Alzheimer's disease (AD). Its inhibition is a valid approach to the treatment of AD. In this initial letter, some thienyl and phenyl alpha-halomethyl ketones are described as new non ATP competitive inhibitors of GSK-3beta. They are considered as lead compounds for designing and synthesizing new series, to carry out SAR studies, clear up the mechanism of action, and, in general, evaluate their therapeutical usefulness. PMID- 14561082 TI - Inclusion complexation and solubilization of paclitaxel by bridged bis(beta cyclodextrin)s containing a tetraethylenepentaamino spacer. AB - A novel water-soluble paclitaxel complex has been prepared by inclusion complexation with bridged bis(beta-cyclodextrin)s and characterized by means of (1)H NMR, SEM, powder X-ray diffraction patterns, TG-DTA, DSC, FT-IR, and 2D NOESY. The cyclodextrins were able to solubilize paclitaxel to levels as high as 2 mg/mL. Furthermore, the cytotoxicity of the novel complexes was assessed using a K562 leukemia cell line which indicated that drug concentrations of 10 pg/mL elicited an inhibitory effect. PMID- 14561083 TI - Kinases, homology models, and high throughput docking. AB - With the many protein sequences coming from the genome sequencing projects, it is unlikely that we will ever have an atomic resolution structure of every relevant protein. With high throughput crystallography, however, we will soon have representative structures for the vast majority of protein families. Thus the drug discovery and design process will rely heavily on protein modeling to address issues such as designing combinatorial libraries for an entire class of targets and engineering genome-wide selectivity over a target class. In this study we assess the value of high throughput docking into homology models. To do this we dock a database of random compounds seeded with known inhibitors into homology models of six different kinases. In five of the six cases the known inhibitors were found to be enriched by factors of 4-5 in the top 5% of the overall scored and ranked compounds. Furthermore, in the same five cases the known inhibitors were found to be enriched by factors of 2-3 in the top 5% of the scored and ranked known kinase inhibitors, thus showing that the homology models can pick up some of the crucial selectivity information. PMID- 14561084 TI - Development of potent inhibitors of botulinum neurotoxin type B. AB - Botulinum neurotoxins are the most potent toxins known to date. They are zinc metalloproteases able to cleave selectively an essential component of neurotransmitter exocytosis, causing the syndrome of botulism characterized by a flaccid paralysis. There is a great interest in designing antagonists of the action of these toxins. One way is to inhibit their catalytic activity. In this study, we report the design of such inhibitors directed toward BoNT/B. A study of the S(1) subsite specificity, using several beta-amino thiols, has shown that this subsite prefers a p-carboxybenzyl moiety. The specificity of the S(1)' and S(2)' subsites was studied using two libraries of pseudotripeptides containing the S(1) synthon derived from the best beta-amino thiol tested. Finally, a selection of various non natural amino acids for the recognition of the "prime" domain led to the most potent inhibitor of BoNT/B described to date with a K(i) value of 20 nM. PMID- 14561085 TI - Receptor-guided alignment-based comparative 3D-QSAR studies of benzylidene malonitrile tyrphostins as EGFR and HER-2 kinase inhibitors. AB - The overexpression and/or mutation of the epidermal growth factor receptor family of tyrosine kinases, namely EGFR and HER-2, have been implicated in poor prognosis of human solid tumors and are under investigation as molecular targets for cancer therapy. To gain insights into selectivity in the interaction of inhibitors at the ATP site of the two kinases, we have carried out docking, comparative molecular field analysis (CoMFA), and comparative molecular similarity analysis (CoMSIA) 3D-QSAR studies on 50 benzylidene malonitrile derivatives. Docking studies indicate different binding modes (A, B, C, D, and E) that are dependent on the R(1) substituent of the compounds. Binding modes A and B are favored by compounds having a hydroxyl substituent at R(1) whereas a methoxy substituent at R(1) results in compounds occupying binding modes, C, D, and E. The compounds preferred modes A, B, and C in the apo-enzyme whereas modes B, D, and E were preferred in the enzyme in complex with erlotinib. For 3D QSAR studies, based on the multiple binding modes obtained from the docking, four composite alignment strategies (I, II, III, IV) were employed and compared with alignment V, which is based on pairwise atom alignment of the common structural elements. Alignments I and II produced models with better predictive ability than those from alignments III and IV against an external test set. In the EGFR kinase results, alignments I and II produced comparable 3D-QSAR models with alignment II being slightly better than I, whereas in the HER-2 results, alignment I was better than alignment II in its predictive ability. It appears that differences in binding mode preferences at the ATP site might constitute a reason for the selectivity of the dihydroxy compounds as inhibitors of EGFR relative to HER-2. They are more likely to have multiple binding modes at the ATP site of EGFR, i.e., either modes A or B, than in the ATP site of HER-2 where they are possibly limited to only binding mode, A. Selectivity of the methoxy compounds on the other hand appears to depend on hydrogen bonding interactions involving the cyano group and residue 751 in the ATP site. PMID- 14561087 TI - Structure-activity relationships of the p38alpha MAP kinase inhibitor 1-(5-tert butyl-2-p-tolyl-2H-pyrazol-3-yl)-3-[4-(2-morpholin-4-yl-ethoxy)naph- thalen-1 yl]urea (BIRB 796). AB - We report on the structure-activity relationships (SAR) of 1-(5-tert-butyl-2-p tolyl-2H-pyrazol-3-yl)-3-[4-(2-morpholin-4-yl-ethoxy)naphthalen-1-yl]urea (BIRB 796), an inhibitor of p38alpha MAP kinase which has advanced into human clinical trials for the treatment of autoimmune diseases. Thermal denaturation was used to establish molecular binding affinities for this class of p38alpha inhibitors. The tert-butyl group remains a critical binding element by occupying a lipophilic domain in the kinase which is exposed upon rearrangement of the activation loop. An aromatic ring attached to N-2 of the pyrazole nucleus provides important pi CH(2) interactions with the kinase. The role of groups attached through an ethoxy group to the 4-position of the naphthalene and directed into the ATP-binding domain is elucidated. Pharmacophores with good hydrogen bonding potential, such as morpholine, pyridine, and imidazole, shift the melting temperature of p38alpha by 16-17 degrees C translating into K(d) values of 50-100 pM. Finally, we describe several compounds that potently inhibit TNF-alpha production when dosed orally in mice. PMID- 14561086 TI - Thermal denaturation: a method to rank slow binding, high-affinity P38alpha MAP kinase inhibitors. AB - It has been reported that the diaryl urea class of p38alpha inhibitors binds to p38 map kinase with both high affinity and slow binding kinetics (Pargellis et al. Nat. Struct. Biol. 2002, 9, 268-272). The slow binding kinetics of this class of inhibitors is believed to be the result of binding to an allosteric pocket adjacent to the p38alpha active site. The use of traditional kinetic and equilibrium methods to measure the binding affinity of this class of compounds has created many challenges for determination of structure-activity relationships (SAR). The thermal denaturation method provides a means of measuring high affinity interactions. In this paper, the method of thermal denaturation will be described as it has been applied to the diaryl urea class of p38 map kinase inhibitors. PMID- 14561088 TI - Molecular determinants of steroid inhibition for the mouse constitutive androstane receptor. AB - The constitutive androstane receptor (CAR) regulates drug and steroid metabolism through binding to cytochrome P450 2B, 2C, and 3A gene enhancers. Uniquely among nuclear receptors, mouse CAR (mCAR) can be suppressed by androstenol and activated by structurally diverse drugs, pesticides, and environmental pollutants. To gain insight into presently ill-defined structural requirements of mCAR ligands, we employed a mCAR inhibition assay in mammalian HEK293 cells to create a QSAR model that could well predict the inhibition by three unknown steroids. Two novel mCAR inhibitors were thus identified. Yeast two-hybrid assays indicated that steroids inhibit mCAR primarily by promoting association of mCAR with the corepressor NCoR, with only minor contribution from other mechanisms. Analysis of chimeric and mutant mCAR constructs suggested that androstenol sensitivity is controlled by residues between amino acids 201-263 (helices 5-7) and it does not depend on the residue 350 within helix 12, as previously suggested. PMID- 14561089 TI - Design, synthesis, and crystal structure of selective 2-pyridone tissue factor VIIa inhibitors. AB - Targeted 2-pyridones were selected as tissue Factor VIIa inhibitors and prepared from 2,6-dibromopyridine via a multistep synthesis. A variety of chemical transformations, including regioselective nucleophilic addition, selective nitrogen alkylation, and a Suzuki coupling, afforded the targeted tissue Factor VIIa inhibitors. The pyridone core was selected as a replacement for the pyrazinone core of noncovalent tissue Factor VIIa inhibitors and designed such that their substitution pattern would occupy and interact with the S(1), S(2), and S(3) pockets of the tissue Factor VIIa enzyme. These compounds were tested in several serine protease enzyme assays involved in the coagulation cascade exhibiting modest activity on tissue Factor VIIa with excellent selectivity over thrombin and Factor Xa. Finally, an X-ray crystal structure of inhibitor 14a bound to tissue Factor VIIa was obtained and will be described. PMID- 14561090 TI - Design and synthesis of 4-azaindoles as inhibitors of p38 MAP kinase. AB - Inhibition of the biosynthesis of proinflammatory cytokines such as tumor necrosis factor and interleukin-1 via p38 has been an approach toward the development of a disease modifying agent for the treatment of chronic inflammation and autoimmune diseases. The development of a new core structure of p38 inhibitors, 3-(4-fluorophenyl)-2-(pyridin-4-yl)-1H-pyrrolo[3,2-b] pyridine, is described. X-ray crystallographic data of the lead bound to the active site of p38 was used to guide the optimization of the series. Specific focus was placed on modulating the physical properties of the core while maintaining potent inhibition of p38. These efforts identified 42c as a potent inhibitor of p38, which also possessed the required physical properties worthy of advanced studies. PMID- 14561091 TI - Azasterols as inhibitors of sterol 24-methyltransferase in Leishmania species and Trypanosoma cruzi. AB - This paper describes the synthesis of some novel azasterols based on (20R,22xi) 5alpha-pregnan-20-(piperidin-2-yl)-3beta,20-diol. These compounds are potential inhibitors of the enzyme sterol 24-methyltransferase (24-SMT), which is a vital enzyme in the biosynthesis of ergosterol and related 24-alkyl sterols. Structure activity studies were undertaken to understand the important features for activity against the enzyme, with the aim of increasing activity and selectivity. The compounds were evaluated for inhibition of recombinant Leishmania major 24 SMT and the effect of compounds on sterol composition and parasite proliferation. Essentially, compounds which showed good activity against the recombinant enzyme had a significant effect on the sterol composition and growth of parasites. The activity of compounds was found to be related to the basicity and stereochemical location of the nitrogen. Also, presence of an unprotected 3beta-OH seemed to be important for activity. However, some azasterols which were not good inhibitors of 24-SMT also showed antiproliferative activity, suggesting that there may be other modes of actions of these compounds. PMID- 14561092 TI - Synthesis and pharmacological evaluation of 1-oxo-2-(3-piperidyl)-1,2,3,4- tetrahydroisoquinolines and related analogues as a new class of specific bradycardic agents possessing I(f) channel inhibitory activity. AB - A series of 1-oxo-2-(3-piperidyl)-1,2,3,4-tetrahydroisoquinolines and related analogues were prepared and evaluated for their bradycardic activities in isolated right atrium and in anesthetized rats. (+/-)-6,7-Dimethoxy-2-[1-[3-(3,4 methylenedioxyphenoxy)propyl]-3-piperidyl]-1,2,3,4-tetrahydroisoquinoline (4) was chosen as a lead, and structural modifications were performed on the tetrahydroisoquinoline ring and the terminal aromatic ring. The modifications on the tetrahydroisoquinoline ring revealed that the 1-oxo-1,2,3,4 tetrahydroisoquinoline ring system was optimum structure for both in vitro potency and in vivo efficacy. Furthermore, methoxy, ethoxy, and methoxycarbonyl groups were identified as preferable substituents on the terminal aromatic ring. One of the 1-oxo-1,2,3,4-tetrahydroisoquinoline derivatives, (R)-10a, was further evaluated for its bradycardic activity and inhibitory activity against I(f) currents. Compound (R)-10a demonstrated potent bradycardic activity in rats with minimal influence on blood pressure after oral administration. The compound also showed inhibition of I(f) currents (IC(50) = 0.32 muM) in guinea pig pacemaker cells. PMID- 14561093 TI - Convergent synthesis and unexpected Ca(2+)-mobilizing activity of 8-substituted analogues of cyclic ADP-carbocyclic-ribose, a stable mimic of the Ca(2+) mobilizing second messenger cyclic ADP-ribose. AB - Cyclic ADP-carbocyclic-ribose (cADPcR, 2) is a biologically and chemically stable equivalent of cyclic ADP-ribose (cADPR, 1), a Ca(2+)-mobilizing second messenger. In this study, a series of 8-substituted analogues of cADPcR, namely the 8-chloro analogue 6 (8-Cl-cADPcR), the 8-azido analogue 7 (8-N(3)-cADPcR), the 8-amino analogue 8 (8-NH(2)-cADPcR), and the 8-phenylthio analogue 9 (8-SPh-cADPcR), were designed as effective pharmacological tools for studies on cADPR-modulated Ca(2+) signaling pathways. These target compounds were synthesized by a convergent route via 8-Cl-cADPcR bisacetonide (14) as the common intermediate, in which a method for forming the intramolecular pyrophosphate linkage by activation of the phenylthiophosphate type substrate 15 with AgNO(3) to produce 14 was used as the key step. The carbocyclic analogues were tested for activity in the sea urchin egg homogenate system. Compounds were assessed for their calcium-mobilizing effects and their ability to cross-desensitize with calcium release induced by a normally maximal concentration of cADPR, as well as cADPR antagonism of cADPR evoked calcium release. While cADPcR was 3-4 times more potent than cADPR, the 8 substituted analogues were less efficacious, with 8-SPh-cADPcR largely acting as a competitive antagonist. Most surprisingly, given that 8-N(3)-cADPR and 8-NH(2) cADPR are known as potent antagonists, 8-N(3)-cADPcR and 8-NH(2)-cADPcR were full agonists, but ca. 80 and 2 times less potent than cADPR, respectively. These data contribute to developing structure-activity relationships for the interaction of cADPR with its receptor. PMID- 14561094 TI - Adenosine kinase inhibitors. 3. Synthesis, SAR, and antiinflammatory activity of a series of l-lyxofuranosyl nucleosides. AB - Chronic inflammatory diseases, such as arthritis and rheumatoid arthritis, remain major health problems worldwide. We previously demonstrated that adenosine kinase inhibitors (AKIs) exhibit antiinflammatory effects by inhibiting TNF-alpha production, neutrophil accumulation, and edema formation. Although adenosine receptor agonists produce similar effects, AKIs showed the antiinflammatory activity without the cardiovascular side effects that prevented the development of adenosine receptor specific agonists. However, previously described potent AKIs, such as 5-iodotubercidin, are nucleosides which have the potential to undergo in vivo 5'-O-phosphorylation and therefore produce cytotoxicity. In an effort to eliminate toxicities produced by phosphorylated nucleosides, l lyxofuranosyl analogues of tubercidin were tested as potential AKIs since the opposite stereochemical orientation of the CH(2)OH was expected to eliminate intracellular phosphorylation. Described herein are the discovery of a new series of AKIs based on alpha-l-lyxofuranosyl nucleosides, their SAR, as well as the antiinflammatory activity of the lead compound GP790 (IC(50) = 0.47 nM, 47% inhibition of paw swelling at 10 mg/kg in rat carrageenan paw edema model). In addition, a study showing that in the skin lesion model the antiinflammatory activity is reversed by an A2 selective adenosine receptor antagonist 3,7 dimethyl-1-propargyl xanthine [correction of propylxanthine] (DMPX) is also described. PMID- 14561095 TI - Synthesis and antiprotozoal activity of aza-analogues of furamidine. AB - 6-[5-(4-Amidinophenyl)furan-2-yl]nicotinamidine (8a) was synthesized from 6-[5-(4 cyanophenyl)furan-2-yl]nicotinonitrile (4a), through the bis-O-acetoxyamidoxime followed by hydrogenation. Compound 4a was prepared via selective bromination of 6-(furan-2-yl)nicotinonitrile (2a) with N-bromosuccinimide, followed by Suzuki coupling with 4-cyanophenylboronic acid. In a similar way, diamidines 8b and 8c were prepared from the dicyano derivatives 4c and 4d, respectively. N-Methoxy-6 [5-[4-(N-methoxyamidino)phenyl]-furan-2-yl]-nicotinamidine (6a) was prepared via methylation of the respective diamidoxime 5a with dimethylsulfate. Prodrugs 6b and 6c were also prepared by methylation of the respective diamidoximes 5b and 5d. The symmetrical diamidines 14a,b were synthesized through the corresponding bis-O-acetoxyamidoxime followed by hydrogenation. The key compounds 11a,b were conveniently obtained by Stille coupling between 2,5-bis(tri-n-butylstannyl)furan and the corresponding heteroaryl halides. These compounds have been evaluated in vitro for activity against Trypanosoma b.rhodesiense (T. b. r.) and P. falciparum (P. f.). The diamidines 8a, 8c, and 14b gave IC(50) values versus T. b. r. of less than 10 nM. Against P. f. 8a, 8b, and 14b exhibited IC(50) values less than 10 nM. In an in vivo mouse model for T. b. r. four compounds 6a, 6c, 6d, and 8a were curative. Compound 6a produced cures at an oral dosage of 5 mg/kg. PMID- 14561096 TI - Finding discriminating structural features by reassembling common building blocks. AB - We present a new method for constructing discriminating substructures by reassembling common medicinal chemistry building blocks. The algorithm can be parametrized to meet differing objectives: (1) to build features that discriminate for biological activity in a local structural neighborhood, (2) to build scaffolds for R-group analysis, (3) to construct cluster signatures that discriminate for membership in the cluster and provide a graphical representation for its members, and (4) to identify substructures that characterize major classes in a heterogeneous compound set. We illustrated the results of the algorithm on a literature dataset is of 118 compounds with in vitro inhibition data against recombinant human protein tyrosine phosphatase 1B (PTP-1B). PMID- 14561097 TI - Potent inhibition of NTPase/helicase of the West Nile Virus by ring-expanded ("fat") nucleoside analogues. AB - A series of ring-expanded ("fat") nucleoside analogues (RENs) containing the 6 aminoimidazo[4,5-e][1,3]diazepine-4,8-dione ring system have been synthesized and screened for inhibition of NTPase/helicase of the West Nile Virus (WNV). To assess the selectivity of RENs against the viral enzymes, a truncated form of human enzyme Suv3((Delta)(1)(-)(159)) was also included in the study. Ring expanded nucleosides 16, 17, and 19, which possess the long C(12), C(14), and C(18) side-chains, respectively, at position 6, as well as the ring-expanded heterocycle 39, which contains aralkyl substitution at position 1, were all found to have excellent profiles of activity and selectivity toward the viral versus human enzymes against the West Nile Virus (IC(50) ranging 1-10 muM). Compound 30, while being an equally potent inhibitor of WNV, was found to be somewhat less selective, whereas compound 36, which is an alpha-anomeric counterpart of 30, exhibited potent and selective inhibition of WNV (IC(50) 1-3 muM). The same compounds that showed potent inhibition of viral helicase activity completely failed to show any activity against the viral NTPase reaction even up to 500 muM. However, at concentrations >500 muM of RENs and the ATP concentrations >10 times the K(m) value of the enzyme, a significant activation of NTPase activity was observed. This activating effect underwent further dramatic enhancement (>1000%) by further increases in ATP concentration in the reaction mixture, suggesting that the viral helicase and NTPase reactions are not coupled. A tentative mechanistic model has been proposed to explain the observed results. PMID- 14561098 TI - Nonpeptide alphavbeta3 antagonists. 8. In vitro and in vivo evaluation of a potent alphavbeta3 antagonist for the prevention and treatment of osteoporosis. AB - 3(S)-(6-methoxypyridin-3-yl)-3-[2-oxo-3-[3-(5,6,7,8-tetrahydro-[1,8]-naphthyridin 2-yl)propyl]imidazolidin-1-yl]propionic acid 6 was identified as a potent and selective antagonist of the alpha(v)beta(3) receptor. This compound has an excellent in vitro profile (IC(50) = 0.08 nM), a significant unbound fraction in human plasma (12%), and good pharmacokinetics in rat, dog, and rhesus monkey. On the basis of the efficacy shown in three in vivo models of bone turnover, the compound was selected for clinical development. To support the ongoing metabolism and safety studies, a novel strategy was employed in which a series of oxidized derivatives of 6 were prepared by exposure of 6 (or the methyl ester) to chemical oxidizing agents. These products proved useful in the identification of active metabolites generated by either in vitro or in vivo metabolism. PMID- 14561099 TI - Inhibition of human immunodeficiency virus reverse transcriptase by synadenol triphosphate and its E-isomer. AB - Triphosphate 1c is a potent competitive inhibitor of wild-type HIV-1 reverse transcriptase with K(i) close to ddATP. The E-isomer 2c is about 30-times weaker. Triphosphates 1c and 2c interact with the same active site of reverse transcriptase as ddATP. The extent of inhibition of two mutant forms of reverse transcriptase (RT), RT(M184V) and RT(M184I), with triphosphate 1c was about 5 and 8 times lower than that of wild-type RT(wt). PMID- 14561100 TI - Pharmacology and therapeutic effects of dehydroepiandrosterone in older subjects. AB - Man and higher primates have adrenals that secrete large amounts of dehydroepiandrosterone (DHEA) [prasterone] and its sulphate (DHEAS) [PB 008]. A remarkable feature of plasma DHEAS levels in humans is their great decrease with aging. Researchers have postulated that this age-related decline of DHEAS levels may explain some of the degenerative changes associated with aging. Moreover, administration of DHEA to laboratory animals has demonstrable beneficial effects such as prevention of diabetes mellitus, obesity, cancer, heart disease and positive immunomodulator effects. However, in rodents DHEA(S) circulating levels are so low that it is impossible to detect any significant age-related decrease. Therefore results from rodent experiments are not relevant to human beings. Three mechanisms of action of DHEA(S) have been identified. DHEA and DHEAS are precursors of testosterone and estradiol, DHEAS is a neurosteroid which modulates neuronal excitability via specific interactions with neurotransmitter receptors and DHEA is an activator of calcium-gated potassium channels. Randomised, placebo controlled clinical trials which included healthy individuals aged 60 years and over treated with (near) physiological doses of DHEA (50-100 mg/day) have yielded very few positive results. Impact of DHEA replacement treatment was assessed on mood, well being, cognitive and sexual functions, bone mass, body composition, vascular risk factors, immune functions and skin. The major limitations of these trials were their short duration (maximum 1 year) and the low number of study participants involved (maximum 280). Many elderly people in western countries take DHEA without medical supervision. In the US, DHEA is even classified as food supplement. At present there is no scientific evidence to recommend DHEA replacement in the elderly. Further studies are needed to form conclusions about the efficacy and the safety of DHEA replacement in elderly, and to better understand the mechanisms of action of DHEA at the molecular and cellular levels. PMID- 14561101 TI - Clinically important drug interactions with intravenous anaesthetics in older patients. AB - There is a continuously growing population of older surgical patients who require an increasing number of anaesthetics and sedation. Pharmacokinetic and pharmacodynamic changes associated with increasing age are often not appreciated enough. Dose requirements for midazolam, a benzodiazepine commonly used for outpatient procedures, have been demonstrated in prospective studies to decrease with increasing age. On the other hand, rigorous prospective studies investigating the effect of age on the induction doses of other intravenous anaesthetics, such as thiopental sodium or propofol, are missing. In addition, many of those patients take multiple drugs for medical problems often not related to the procedure. Drug interactions with anaesthetics are likely to occur, but are not well documented. In this review we have summarised the documented and clinically relevant drug interactions with anaesthetics in the elderly population. We have identified a significant lack of scientific and outcome data and the need for more studies and education. PMID- 14561106 TI - Parental vocalizations and perceived immaturity in down syndrome. AB - Previous findings suggest that children with Down syndrome may be perceived as more youthful than other children with mental retardation. In this study whether perceptions of youthfulness can be linked to parenting behavior, in particular increased prosodic adjustments characteristic of infant-directed speech, is investigated. The prosodic characteristics of parental language targeted to children with Down syndrome and children with other etiologies of mental retardation were compared. Results show that parents of children with Down syndrome raised their voice pitch significantly more (i.e., spoke in a higher register) and spoke with a significantly wider mean pitch variance than parents of children in the comparison group. Implications are discussed for parent-child interactions in children with different genetic mental retardation syndromes. PMID- 14561103 TI - Aspirin for the prevention of cardiovascular events in the elderly. AB - Aspirin (acetylsalicylic acid), the most widely used antiplatelet drug, is clinically effective for the prevention of vascular ischaemic events. Very few primary or secondary prevention trials address the benefit-risk ratio of aspirin in the elderly. In secondary prevention, it is generally accepted that the beneficial effect of aspirin in the general patient population, demonstrated by randomised controlled trials, can be extrapolated to the elderly. Elderly patients are at relatively high risk for the development of vascular disease and might also be expected to derive substantial benefit from regular aspirin administration. However, there is no consensus about the definition of elderly and no specific prospective trial conducted in elderly subjects is available. Retrospective studies in the elderly found that the benefit provided by aspirin in older patients was similar or increased compared with younger individuals. In primary prevention, the potential benefit of antiplatelet agents must be balanced against the risk of bleeding, which is higher in older patients. The risk-benefit trade-off from the use of low-dose aspirin in the elderly is not yet established and caution should be exercised when using aspirin in primary prevention. In conclusion, aspirin should only be given for primary and secondary prevention in the elderly after a comprehensive evaluation of an individual patient's thrombotic and haemorrhagic risk has been conducted. PMID- 14561102 TI - Complementary and alternative medicines in the treatment of dementia: an evidence based review. AB - Alternative medicines may have potential beneficial results in treating certain forms of dementia and related symptoms, as well as slowing disease progression. Alternative medicines may ameliorate disturbances in cognition, mood, sleep and activities of daily living. Primary mechanisms of action include modifications in neurotransmitter synthesis, inhibition of neurotransmitter reuptake and enzyme induced neurotransmitter breakdown, antioxidant and anti-platelet activity, enhanced blood flow and glucose metabolism. Adverse events can include cardiovascular, gastrointestinal, mood, autonomic and dermatologic effects. However, adverse events, when reported represent, a small percentage of treated groups and direct links between adverse events and alternative therapies are tenuous. Many studies of alternative medicines in dementia are inconclusive and characterised by methodological deficiencies such as small sample sizes and inadequate controls. If alternative medicines can be shown to be efficacious using more rigorous experimental designs, both consumers and clinicians could avail themselves of a wider range of pharmacological substances that may offer the advantage of being better tolerated and exhibiting safer therapeutic margins than some allopathic medicines. While a number of complementary interventions have shown both strengths and weaknesses, huperzine A, levacecarnine and EGB 761, based on the overall quality of the studies, identified mechanisms of activity and safety profiles merit further examination in controlled clinical outcome studies. PMID- 14561107 TI - beta-Endorphin and ACTH are dissociated after self-injury in adults with developmental disabilities. AB - Relations between self-injuring behavior (SIB), the hypothalamic-pituitary adrenal (HPA) stress axis, and response to an opiate antagonist were examined. Subjects were observed in their residential settings, while behavior was recorded. Blood was collected in the morning, evening, and immediately after SIB. Plasma beta-E was uncoupled from ACTH after SIB but not during the morning baseline. A significant number of the subjects (a) reduced their SIB at least 25% at all doses of naltrexone (NTX) and (b) reduced their SIB over 50% for at least one dose of NTX. The lowest dosage of NTX significantly reduced SIB in subjects with baseline levels of beta-E higher than after SIB. Results support previous reports that the HPA axis is disturbed among subjects exhibiting SIB. PMID- 14561108 TI - Phonological and visuo-spatial working memory in individuals with intellectual disability. AB - Differences in the storage and rehearsal components of the phonological loop and visuo-spatial sketchpad were investigated in individuals with and without intellectual disability matched on memory span. The group with intellectual disability had specific difficulty in the rehearsal component of the phonological loop, as demonstrated by a weak word length effect compared to the group without intellectual disability. Groups did not differ in the storage component of the phonological loop as indexed by the phonological similarity effect. Also, groups did not differ in the storage or maintenance components of the visuo-spatial sketchpad, having comparable visual similarity and visual complexity effects. However, visual complexity task performance suggested that some aspects of visual processing surpass developmental level expectations for individuals with intellectual disability. PMID- 14561109 TI - Structural and environmental characteristics of stereotyped behaviors. AB - Structural and environmental characteristics of multiple topographies of stereotypic behaviors shown by 8 individuals with developmental disabilities are described. Analysis of structural characteristics show that the parameters of stereotyped behavior (percentage of time, bout length, and bout length variability) were highly correlated. Analysis of the influence of environmental variables indicated that topographies of stereotyped behaviors were more likely to occur under conditions of low stimulation and less likely during conditions involving social contact. Individuals whose stereotyped behaviors were less influenced by environmental factors were more likely to engage in them for greater periods of time. Results suggest that an examination of individual topographies of stereotyped behaviors and their structural and environmental characteristics may provide useful insights for understanding the origins of these behaviors. PMID- 14561110 TI - Adaptive behavior in children with fragile X syndrome. AB - Adaptive behavior over time in 70 children with fragile X syndrome, ages 1 to 12 years, was examined using the Vineland Adaptive Behavior Scales. With a mean of 4.4 assessments per child, adaptive behavior skills increased steadily and gradually over time. Children with less autistic behavior and higher percentages of FMPR expression showed better performance on all areas of adaptive behavior. Children without autistic behavior displayed higher scores and rates of growth on the Daily Living Skills domain, with the lowest scores in Socialization. Comparison to Brief IQs indicate that children with fragile X syndrome display nonverbal IQs superior to their adaptive behavior when they are below age 10 but that these skills seem to converge as they get older. PMID- 14561111 TI - Effects of donepezil on cognitive functioning in Down syndrome. AB - Our goal in this study was to determine whether donepezil, an acetylcholinesterase inhibitor, would improve cognitive functioning in 19 subjects with Down syndrome and no dementia. They were assigned to either a donepezil or placebo group. Cognitive functioning and caregiver ratings were measured at baseline, 4 weeks, and 12 weeks. With the exception of one area (language), no improvement was noted in any of the cognitive subtests, behavioral scores, or caregiver ratings. Subjects in the donepezil group showed an improvement in language scores compared to subjects in the placebo group. The results suggest that donepezil may improve language performance in subjects with Down syndrome and no dementia, but further studies need to be done on a larger group to confirm this result. PMID- 14561112 TI - The effect of human immunodeficiency virus-1 protease inhibitors on the toxicity of a variety of cells. AB - Protease inhibitors in combination with other antiretroviral drugs have been shown to be efficacious in treating human immunodeficiency virus-1 (HIV-1) infection. The side effects of such a treatment usually involve perturbations of fat metabolism and insulin responsiveness. This has led to a number of studies on the adverse effects of these drugs in vitro. The concentrations of various protease inhibitors used in many of these studies were >20 microM. Although some investigators did address the toxicity of protease inhibitors, no overall effort was made to examine this effect during differentiation of fat or muscle. In this study, we assessed the toxicity of HIV-1 protease inhibitors over a range of concentrations (i.e., 0 to 100 microM) in nondifferentiating (e.g., human fibroblasts, 3T3-L1 preadipocytes, and L6 myoblasts) and differentiated cells (e.g., L6 myotubes). The most toxic protease inhibitor in all cell types was Saquinavir (sqv), whereas the least toxic protease inhibitor was Indinavir (idv). Ritonavir (rtv) and Amprenavir (apv) were more toxic than idv but not quite as toxic as sqv. In 3T3-L1 preadipocytes, treatment with sqv, rtv, and apv resulted in toxicity, whereas idv was not toxic even at the highest concentration used. Indinavir was not toxic to L6 myoblasts or L6 myotubes; however, sqv, rtv, and apv caused toxicity in L6 myoblasts. Saquinavir decreased L6 myotube viability in a dose-dependent manner. Human immunodeficiency virus-1 protease inhibitors were shown to be toxic in a variety of cell types. These effects on human fibroblasts and muscle cells have not been reported previously. PMID- 14561113 TI - Introduction to the special section on social neuroscience: promise and caveats. AB - This special issue of the Journal of Personality and Social Psychology: Attitudes and Social Cognition is devoted to theory and research at the interface of social psychology and neuroscience. The 5 empirical articles represent the theoretical and methodological breadth of issues considered by social neuroscientists. The methods span brain lesion work to neuroendocrinology to psychophysiological indicators of brain activity to functional magnetic resonance imaging indicators of brain activity. The remaining 2 articles consider explicitly some of the promises and pitfalls of social neuroscience; these authors, although noting the power of neuroscience methods, remind readers of the serious challenges posed in trying to examine the biological processes underlying or associated with social psychological phenomena. These articles help to reveal the richness of social neuroscience and the power of neuroscientific methods to address processes and mechanisms that would not be possible with traditional social psychology methods. PMID- 14561114 TI - The regulatory function of self-conscious emotion: insights from patients with orbitofrontal damage. AB - Although once considered disruptive, self-conscious emotions are now theorized to be fundamentally involved in the regulation of social behavior. The present study examined the social regulation function of self-conscious emotions by comparing healthy participants with a neuropsychological population--patients with orbitofrontal lesions--characterized by selective regulatory deficits. Orbitofrontal patients and healthy controls participated in a series of tasks designed to assess their social regulation and self-conscious emotions. Another task assessed the ability to infer others' emotional states, an appraisal process involved in self-conscious emotion. Consistent with the theory that self conscious emotions are important for regulating social behavior, the findings show that deficient behavioral regulation is associated with inappropriate self conscious emotions that reinforce maladaptive behavior. Additionally, deficient behavioral regulation is associated with impairments in interpreting the self conscious emotions of others. PMID- 14561115 TI - Are self-enhancing cognitions associated with healthy or unhealthy biological profiles? AB - Self-enhancement is variously portrayed as a positive illusion that can foster health and longevity or as defensive neuroticism that can have physiological neuroendocrine costs. In a laboratory stress-challenge paradigm, the authors found that high self-enhancers had lower cardiovascular responses to stress, more rapid cardiovascular recovery, and lower baseline cortisol levels, consistent with the positive illusions predictions and counter to the predictions of the defensive neuroticism position. A second set of analyses, replicating the "illusory mental health paradigm" (J. Shedler, M. Mayman, & M. Manis, 1993), also did not support the defensive neuroticism hypothesis. The association between self-enhancement and cortisol was mediated by psychological resources; analyses of the cardiovascular results provided no definitive mediational pathway. Discussion centers on the potential stress-buffering effects of self-enhancing beliefs. PMID- 14561116 TI - Race and gender on the brain: electrocortical measures of attention to the race and gender of multiply categorizable individuals. AB - The degree to which perceivers automatically attend to and encode social category information was investigated. Event-related brain potentials were used to assess attentional and working-memory processes on-line as participants were presented with pictures of Black and White males and females. The authors found that attention was preferentially directed to Black targets very early in processing (by about 100 ms after stimulus onset) in both experiments. Attention to gender also emerged early but occurred about 50 ms later than attention to race. Later working-memory processes were sensitive to more complex relations between the group memberships of a target individual and the surrounding social context. These working-memory processes were sensitive to both the explicit categorization task participants were performing as well as more implicit, task-irrelevant categorization dimensions. Results are consistent with models suggesting that information about certain category dimensions is encoded relatively automatically. PMID- 14561117 TI - Effects of alcohol on person perception: a social cognitive neuroscience approach. AB - The acute effects of alcohol on cognitive processing of expectancy violations were investigated using event-related brain potentials and a cued recall task to index attentional and working memory processes associated with inconsistency resolution. As predicted, expectancy-violating behaviors elicited larger late positive potentials (LPP) and were recalled better than expectancy-consistent behaviors. These effects were moderated by alcohol and the valence of initial expectancies. For placebo group participants, positive targets performing negative behaviors elicited the largest LPP responses and were recalled best. For those in the alcohol groups, negative targets behaving positively elicited the largest LPP and recall responses. These findings suggest that alcohol does not globally impair working memory processes in person perception but instead changes the nature of valenced information processing. Findings are discussed in the context of alcohol's effects on working memory processes, reward sensitivity, and the prefrontal cortical structures thought to mediate them. PMID- 14561118 TI - Neural components of social evaluation. AB - Evaluative responses appear to involve 2 seemingly distinct sets of processes: those that are automatically activated and others that are more consciously controlled. Using functional magnetic resonance imaging, the authors investigated the brain systems associated with automatic and controlled evaluative processing. Participants made either evaluative (good-bad) or nonevaluative (past-present) judgments about famous names. Greater amygdala activity was observed for names rated as "bad" relative to those rated as "good," regardless of whether the task directly involved an evaluative judgment (good-bad) or not (past-present). Good bad judgments resulted in greater medial and ventrolateral prefrontal cortex (PFC) activity than past-present judgments. Furthermore, there was greater ventrolateral PFC activity in good-bad judgments marked by greater ambivalence. Together, these findings indicate a neural distinction between processes engaged for automatic and controlled evaluation. Whereas automatic processes are sensitive to simple valence, controlled processes are sensitive to attitudinal complexity. PMID- 14561119 TI - Just because you're imaging the brain doesn't mean you can stop using your head: a primer and set of first principles. AB - Developments within the neurosciences, cognitive sciences, and social sciences have contributed to the emergence of social neuroscience. Among the most obvious contemporary developments are brain-imaging procedures such as functional magnetic resonance imaging. The authors outline a set of first principles designed to help make sense of brain-imaging research within the fields of cognitive and social neuroscience. They begin with a principle few would debate- that social cognition, emotion, and behavior involve the brain--but whose implications might not be entirely obvious to those new to the field. The authors conclude that (a). complex aspects of the mind and behavior will benefit from yet a broader collaboration of neuroscientists, cognitive scientists, and social scientists, and (b). social psychologists bring important theoretical, methodological, and statistical expertise to this interdisciplinary enterprise. PMID- 14561120 TI - What neuroimaging and brain localization can do, cannot do and should not do for social psychology. AB - Interest in bridging social psychology and neuroscience has seen a significant upsurge. Much of this interest has centered on brain localization--the attempt to relate psychological events to locations of brain events. Although many articles have sought to localize brain activity that supports social behavior, scant attention has been paid to the specific methods to be used in integrating brain localization data into psychological theory. The authors describe 4 strategies psychologists can use to integrate brain localization data and psychological theory, and they consider whether social psychology presents special considerations in the use of these strategies. They conclude that brain localization offers a useful tool for some but not all problems in social psychology, and they discuss the types of problems for which it may and may not prove useful. PMID- 14561121 TI - From self-prediction to self-defeat: behavioral forecasting, self-fulfilling prophecies, and the effect of competitive expectations. AB - Four studies explored behavioral forecasting and the effect of competitive expectations in the context of negotiations. Study 1 examined negotiators' forecasts of how they would behave when faced with a very competitive versus a less competitive opponent and found that negotiators believed they would become more competitive. Studies 2 and 3 examined actual behaviors during a negotiation and found that negotiators who expected a very competitive opponent actually became less competitive, as evidenced by setting lower, less aggressive reservation prices, making less demanding counteroffers, and ultimately agreeing to lower negotiated outcomes. Finally, Study 4 provided a direct test of the disconnection between negotiators' forecasts for their behavior and their actual behaviors within the same sample and found systematic errors in behavioral forecasting as well as evidence for the self-fulfilling effects of possessing a competitive expectation. PMID- 14561122 TI - Groups perform better than the best individuals on letters-to-numbers problems: informative equations and effective strategies. AB - One-hundred 3-person groups and 300 individuals solved 2 letters-to-numbers problems, requiring identification of the coding of 10 letters to 10 numbers by proposing an equation in letters, receiving the answer in letters, proposing a hypothesis, and receiving feedback on the hypothesis on each trial. There were 5 instruction conditions: (a). standard, (b). use at least 3 letters on all equations, (c). use at least 4 letters on all equations, (d). number 1 known before beginning problem, and (e). number 9 known before beginning problem. The groups had fewer trials to solution, proposed more complex equations, and identified more letters per equation than the best individuals. Performance was best under instructions to use at least 4 letters and with the number 9 known. PMID- 14561123 TI - Factor structure of English-language personality type-nouns. AB - Although type-nouns (e.g., idiot, hero) are important in person description, lexical studies of personality have concentrated on adjectives. This study tested structural hypotheses using 372 highly familiar English-language type-nouns and descriptions by 607 participants of either themselves, a liked, or a disliked target person. One- and 2-factor structures were most robust, and replicated similar structures found in previous adjectival studies. Additionally, the structure with 8 orthogonal factors had good replicability and applicability within single-gender subsamples; as in previous studies of type-nouns, it included factors corresponding directly to Extraversion and Intellect/Openness, but also to Attractiveness and Masculinity (or Ruggedness). The Big Five was only weakly replicated. Personality taxonomies based on adjectives are unlikely to be comprehensive, because type-nouns have different content emphases. PMID- 14561124 TI - Interpersonal attraction and personality: what is attractive--self similarity, ideal similarity, complementarity or attachment security? AB - Little is known about whether personality characteristics influence initial attraction. Because adult attachment differences influence a broad range of relationship processes, the authors examined their role in 3 experimental attraction studies. The authors tested four major attraction hypotheses--self similarity, ideal-self similarity, complementarity, and attachment security--and examined both actual and perceptual factors. Replicated analyses across samples, designs, and manipulations showed that actual security and self similarity predicted attraction. With regard to perceptual factors, ideal similarity, self similarity, and security all were significant predictors. Whereas perceptual ideal and self similarity had incremental predictive power, perceptual security's effects were subsumed by perceptual ideal similarity. Perceptual self similarity fully mediated actual attachment similarity effects, whereas ideal similarity was only a partial mediator. PMID- 14561125 TI - Relationships between personality structure, structure of word meaning, and cognitive ability: a study of cultural mechanisms of personality. AB - Native-born Estonian men (N=912), 17-68 years old, participated in a study on relationships between personality characteristics, dominant structure of word meaning ("everyday concepts" thinking or "scientific concepts" thinking), and level of cognitive ability. Individuals who primarily used everyday concepts thinking or who possessed relatively low levels of cognitive ability did not reveal a coherent Big Five personality structure, whereas individuals who primarily used scientific concepts thinking or possessed high levels of cognitive ability did. Thus, personality may be shaped by a cultural factor--word meaning structure. Earlier studies, which seem to support the idea that Big Five personality structure is a biologically determined human universal, suffer from serious sampling problems and insufficient data analyses. PMID- 14561126 TI - Confirmatory bias in the evaluation of personality descriptions: positive test strategies and output interference. AB - The operation of confirmatory bias in the endorsement of personality descriptions was examined in 4 studies. Unlike M. F. Davies (1997), who provided only inferential evidence for the role of this bias, the present studies provided direct evidence through the experimental manipulation of supporting versus contradictory cognitions. Generating supporting thoughts resulted in greater acceptance whereas generating contradictory thoughts resulted in lower acceptance of personality descriptions. Supporting cognitions were found to be generated before contradictory cognitions in line with a positive test strategy but evidence was also found for an output interference effect (generating one type of thought interfered with the generation of the opposite type) and it was suggested that confirmatory bias is due to the operation of both mechanisms. PMID- 14561127 TI - Prospective relations between rejection and depression in young adolescents. AB - The purpose of this longitudinal study was to examine the relations between rejection and depression across 3 years in young adolescents who varied with regard to their risk for depression. The sample consisted of 240 adolescents who were assessed in grades 6, 7, and 8. The assessment of rejection was based on adolescent-, mother-, and teacher-report, and depression assessment was based on adolescent- and mother-report and clinician ratings. Structural equation modeling indicated that rejection prospectively predicted depression. The authors did not find that depression prospectively predicted rejection, but such a relation cannot be ruled out because of strong cross-sectional correlations between depression and rejection. PMID- 14561129 TI - The dynamics of masculine-agentic and feminine-communal traits: findings from a prospective study. AB - A reciprocal impact hypothesis posits an influence of gender-related traits (agency and communion) on role enactment and a reciprocal impact of role enactment on gender-related traits, for both men and women. Specifically, in this study it was predicted that agency influences career success and career success influences agency. In addition, the reciprocal influence of communion and family roles was examined. A prospective study with almost 2000 university graduates, who were tested after graduation and 1.5 years later, clearly supported the reciprocal impact hypothesis for agency and career success. Communion influenced family roles, but there was no reciprocal influence. Implications for theories of career success and of sex and gender are discussed. PMID- 14561128 TI - Les passions de l'ame: on obsessive and harmonious passion. AB - Passion is defined as a strong inclination toward an activity that people like, that they find important, and in which they invest time and energy. Two types of passion are proposed: obsessive and harmonious. Obsessive passion (OP) refers to a controlled internalization of an activity in one's identity that creates an internal pressure to engage in the activity that the person likes. Harmonious passion (HP) refers to an autonomous internalization that leads individuals to choose to engage in the activity that they like. HP promotes healthy adaptation whereas OP thwarts it by causing negative affect and rigid persistence. Results from four studies involving more than 900 participants from different populations supported the proposed conceptualization. PMID- 14561130 TI - Synthesis of retro acyl carrier protein (74-65) fragment on a new glycerol based polystyrene support. AB - Retro-ACP (74-65) fragment was synthesized on a novel 4% tri-(propylene glycol) glycerolate diacrylate cross-linked polystyrene (PS-TRPGGDA) support. The peptide is grown from the functional site present in the cross-linker, which makes it unique and cost-effective among other styrene based polymer supports. A comparative study with Merrifield resin indicates high yield and purity of the peptide synthesized on the novel support. PMID- 14561131 TI - A sequence function reveals new features in beta-protein folding. AB - When amino acid residues are represented by parameters describing their side chain lengths and polarities, a sequence function defined as the sum of the first two sequence autocorrelation functions is found to be negatively and linearly correlated with the logarithms of folding rates of beta-proteins. The new function reveals new features in beta-protein folding: larger residues slow down the folding while alternative distribution of polar-non-polar residues accelerates the folding. PMID- 14561132 TI - Multiple roles of glutathione binding-site residues of glutathione s-transferase. AB - This study was designed to characterize residues in the glutathione binding site of AdGSTD4-4 from the mosquito malaria vector Anopheles dirus. The data revealed that Leu33, His38 and His50 each play a role in enzyme catalysis and glutathione binding. The mutants of these three residues also displayed differences in hydrophobic substrate specificity, suggesting that changes in the active site conformation occurred. Differences in conformations was also suggested by protein stability changes. These results indicate that residues in the glutathione binding site are not only important in the catalytic function but also play a role in the structural integrity of the enzyme. PMID- 14561133 TI - A new dehydrogenase specific towards aromatic aldehydes from a halophilic bacterium. AB - A new enzyme showing a dehydrogenase activity towards aromatic aldehydes was isolated, purified and characterized from a halophilic strain isolated from saline environment. The enzyme is a monomer of 54 kDa; it is rather thermostable (optimal temperature: 50 degrees C) showing a broad spectrum of activity in a large pH range with the maximum at pH 9.5. The substrate specificity and the effect of ions were evaluated and compared with analogous described proteins. PMID- 14561134 TI - Direct screening of libraries of yeast clones for alpha-amylase activity on raw starch hydrolysis. AB - High-throughput screening for high-activity barley alpha-amylase mutants expressed in Saccharomyces cerevisiae is hampered by the interference of reducing agents, particularly the glucose used in yeast growth media. The present investigation employed colorimetric and chemiluminescent detection systems that enable direct and rapid screening of activities on raw starch substrate. Active clones could be separated into two groups, based on high total activity or high specific activity. PMID- 14561135 TI - In vivo lipoprotein binding assay of the insect exchangeable apolipoprotein, apolipophorin-III. AB - An original method for the study of the lipid binding properties of exchangeable apolipoproteins is reported. Binding of Locusta migratoria apolipophorin-III to Manduca sexta low-density lipophorin (LDLp) and high-density lipophorin (HDLp) was studied in vivo. This assay could be used useful to investigate the effect of mutations in the lipid binding properties of exchangeable apolipoproteins under physiological conditions. PMID- 14561136 TI - Anticonvulsant activity of benzylamides of some amino acids and heterocyclic acids. AB - A series of new potential anticonvulsants have been synthesized. They are N methyl benzyl-amides of N-methyl Asp and N-methyl Glu (R and S), benzylamides of some heterocyclic acids and their N-oxides and benzylamides of two heteroalicyclic acids. The obtained compounds were evaluated in the Anticonvulsant Screening Project (ASP) of Antiepileptic Drug Development Program (ADDP) of NIH. PMID- 14561137 TI - Extracellular domain of myelin oligodendrocyte glycoprotein (MOG) exhibits solvent-dependent conformational transitions. AB - The conformation of the non-glycosylated recombinant form of the extracellar domain of rat MOG (rMOG(1-125)) dissolved in different solvent conditions was studied by CD spectroscopy. The results show that rMOG(1-125) exhibits a predominantly beta sheet conformation in aqueous buffer solution at pH 7.5 and that this 'beta-form' is stabilized by zwitterionic phospholipids, DPC and LPCP. The alpha helical content of the protein can increase from 9% to up to 20% when TFE or anionic detergent LPAP and SDS are added. PMID- 14561139 TI - Factors determining the efficacy of alpha-helical antimicrobial peptides. AB - A database of alpha-helical antimicrobial peptides (AMP) was established and their minimum inhibitory concentrations (MIC) were compared with their physiochemical characteristics in an attempt to establish those features that determine efficacy. There is no significant difference in AMP sensitivity between Gram-positive and Gram-negative bacteria but fungi did require higher concentrations to achieve the same degree of growth inhibition. For antibacterial peptides there appears to be a positive correlation between MIC and hydrophobic arc size and a negative correlation between MIC and net charge. PMID- 14561138 TI - Regulation of in vitro fibril formation of synuclein mutant proteins by Hsp104p. AB - Hsp104p, as an anti-oxidative protector of ROS generation, was examined to inquire if it prevents aggregation of synuclein mutants, A30P or A53T upon aging, in vitro. The role of Hsp104p was also addressed in dissociation of pre-formed aggregates of synuclein mutants. Significant protection in fibril formation was observed by wild-type Hsp104p regardless of ATP presence, not by mutant Hsp104p. To a lesser extent, the dissociation effect of wild-type Hsp104p was observed only in the presence of ATP. These results will be discussed in relation to the development of an antioxidant approach to prevent amyloid fibril formation in several neurodegenerative diseases. PMID- 14561140 TI - The role of tyrosine residues in the RNA N-glycosidase activity of cinnamomin A chain. AB - Cinnamomin is a type II ribosome-inactivating protein (RIP) and its A-chain (CTA) is a RNA N-glycosidase. It is observed that modification of tyrosine residues by N-acetylimidazole (N-AI) causes almost complete loss of CTA activity. Adenine partially protects tyrosine residues from modification by N-AI. It is proposed that tyrosine residues are involved in the active site of CTA and they are crucial in recognition and binding of ribosomal RNA. Tryptophan residues of CTA are also studied by NBS modification. PMID- 14561142 TI - Crystallization and preliminary X-ray diffraction analysis of phosphoglucose isomerase from Pyrococcus furiosus. AB - In several euryarchaeota, phosphoglucose isomerase (PGI) activity is catalyzed by an enzyme unrelated to the well-known family of PGI enzymes found in prokaryotes, eukaryotes and some archaea. In order to understand the mechanistic differences between the two families of enzymes we have crystallized PGI from the archaeon Pyrococcus furiosus. The crystals belong to the space group P2(1) and a complete dataset extending to 1.9 A resolution has been collected. PMID- 14561141 TI - Crystallization and preliminary X-ray analysis of class II fructose-1,6 bisphosphate aldolase from Thermus caldophilus. AB - In this study, we have crystallized class II fructose-1,6-bisphosphate aldolase (FBA) from Thermus caldophilus (Tca). Purified Tca FBA is a tetrameric enzyme of 305 residues, which crystallizes in the space group P2(1)2(1)2(1) (cell dimensions a = 98.9, b = 113.1, c = 115.7 A), with four molecules in the asymmetric unit. A complete diffraction data set was obtained from orthorhombic crystals at resolution of 2.2 A. PMID- 14561143 TI - Purification, crystallization and preliminary X-ray studies of a p nitrophenylphosphatase from Bacillus stearothermophilus. AB - Thermostable p-nitrophenylphosphatase from Bacillus stearothermophilus has been expressed in Escherichia coli, purified and crystallized. The crystals belong to space group C(2), with unit-cell parameters a = 67.17 A, b = 57.84 A, c = 62.49 A and alpha = 90.0 degrees, beta = 95.4 degrees, gamma = 90.0 degrees. Diffraction data were collected to 1.40 A resolution with a completeness of 94.7% (96.6% for the last shell), an R(fac) value of 0.074 (0.341) and an I/sigma (I) value of 30.1 (2.67). PMID- 14561144 TI - Initiating structural studies of Lys49-PLA2 homologues complexed with an anionic detergent, a fatty acid and a natural lipid. AB - Lys49-Phospholipase A2 (Lys49-PLA(2) - EC 3.1.1.4) homologues damage membranes by a Ca2+-independent mechanism which does not involve catalytic activity. Both MjTX II from Bothrops moojeni and BthTX-I from Bothrops jararacussu are dimeric in solution and in the crystalline states, and a model for the Ca2+-independent membrane damaging mechanism has been suggested in which flexibility at the dimer interface region permits quaternary structural transitions between "open" and "closed" membrane bound dimer conformations which results in the perturbation of membrane phospholipids and disruption of the bilayer structure. With the aim of gaining insights into the structural determinants involved in protein/lipid association, we report here the crystallization and preliminary X-ray analysis of the (i) MjTX-II/SDS complex at a resolution of 2.78A, (ii) MjTX-II/STE complex at a resolution of 1.8 A and (iii) BthTX-I/DMPC complex at 2.72A. These complexes were crystallized by the hanging drop vapour-diffusion technique in (i) HEPES buffer (pH 7.5) 1.8M ammonium sulfate with 2% (w/v) polyethyleneglycol 400, in (ii) 0.6-0.8 M sodium citrate as the precipitant (pH 6.0-6.5) and in (iii) sodium citrate buffer (pH 5.8) and PEG 4000 and 20% isopropanol, respectively. Single crystals of these complexes have been obtained and X-ray diffraction data have been collected at room temperature using a R-AXIS IV imaging plate system and graphite monochromated Cu Kalpha X-ray radiation generated by a Rigaku RU300 rotating anode generator for (i) and (iii) and using using a Synchrotron Radiation Source (Laboratorio Nacional de Luz Sincrotron, LNLS, Campinas, Brazil) for (ii). PMID- 14561146 TI - Hemopoietic progenitor cells and hemopoietic factors: potential targets for treatment of allergic inflammatory diseases. AB - Eosinophilic infiltration is a cardinal feature of allergic inflammation; based upon its biological actions, the eosinophil has assumed the role as the principal inflammatory cell in asthma. In assessing the mechanisms by which eosinophils are recruited to sites of inflammation, a sizeable body of evidence exists supporting the proposal that expansion of hemopoietic compartments in the bone marrow stimulates an increased turnover and traffic of mature eosinophils to the site of allergic inflammation. In addition, recent findings point to the possible egress and traffic of primitive progenitor cells to the site of inflammation where in situ differentiation may provide a continued supply of pro-inflammatory cells. In the present article, we will review the evidence for these findings, and discuss the rationale for targeting hemopoiesis and migrational pathways of hemopoietic cells in the treatment of allergic disease. In this context, we will discuss the effect of corticosteroid treatment on hemopoietic mechanisms; the effects of therapies that inhibit the actions of cysteinyl leukotrienes (CysLTs); the effects of in vivo blockade of the eosinophil-active cytokine, interleukin (IL) 5; and, the effects of antihistamines on hemopoiesis. In addition, we will address the potential role that small molecular weight chemokine receptor antagonists may play in modulating progenitor cell trafficking to tissue sites of inflammation. PMID- 14561147 TI - GM-CSF and dendritic cells in allergic airway inflammation: basic mechanisms and prospects for therapeutic intervention. AB - The interaction between dendritic cells (DC) and naive T cells is the first step in the evolution of an immune response, either tolerogenic or inflammatory. Therefore, the status of DC residing at mucosal sites, such as the airway, has a definitive impact on the character of the ensuing immune response. In the absence of pathogenic stimulation, DC serve to regulate immunological homeostasis in the lung; the generation of Th2-associated (allergic) inflammatory responses, which are directed at presumably innocuous antigens, represent a deviation from normal DC function. The dysregulation of DC phenotype leading to the development of allergy might be programmed by genetic pedigree, or might be induced by factors released in the airway. One potential candidate, GM-CSF, is abundant in the allergic airway and can condition DC to propagate Th2 responses. Moreover, that allergens, alone or in combination with other factors, can spontaneously induce GM-CSF production in the airway thus present a compelling etiological argument for the role of GM-CSF in allergic sensitization. The interplay between DC and mediators present in the allergic airway is likely critical to the establishment of allergic airway inflammation. Understanding these interactions may, therefore, afford insight into prospective therapeutic interventions to circumvent, and even reverse the allergic diathesis. PMID- 14561148 TI - Hemopoietic cells with features of the mast cell and basophil lineages and their potential role in allergy. AB - Human mast cells (MCs) and basophils play a key role in the pathogenesis of allergic disorders, not only by producing inflammatory and fibrogenic mediators, but also by directly and indirectly secreting various cytokines and chemokines. Although mast cells and basophils have differences in many properties, recent evidence suggests that human MCs and basophils may be derived from a common progenitor, and their contents and phenotypes may be reversibly altered in a variety of allergic disorders. The study of FcetaRI signalling of mast cell and basophils offers new opportunities for therapeutic interventions based on the specific inhibition of the earliest events in allergic diseases. This article reviews the origin, differentiation, morphology and phenotypic properties of MCs and basophils, focussing particularly on the possible pathogenic role of MCs and basophils in allergy and biochemical targets for therapeutic interventions in allergic diseases. PMID- 14561149 TI - Mast cell-IgE-and mast cell-structural cell interactions in allergic airway disease. AB - Allergic diseases like atopic rhinitis, bronchial asthma, and urticaria are prevalent and on the rise. Mast cells are known to play a central role in the immediate phase reaction of allergic diseases through the IgE-mediated release of a variety of chemical mediators like histamine, leukotrienes, and prostaglandins. On the other hand, T lymphocytes, basophils and eosinophils are thought to be responsible in inducing the late phase response. Yet, recent studies show that the mast cell cannot be simplistically assigned a role in the immediate phase allergic response, and that this cell plays a crucial role in ongoing allergic inflammation, including the development of hyper-responsiveness. In the present article, the author will try to discuss the integrated roles of mast cells in IgE mediated allergic inflammation with specific emphasis on the roles of mast cell IgE networking and mast cell-structural cell interactions in the late phase allergic response and chronic allergic inflammation. PMID- 14561150 TI - Stem cell factor: a hemopoietic cytokine with important targets in asthma. AB - We review evidence that Stem Cell Factor (SCF) plays an important role in the pathophysiology of asthma. SCF is produced by a wide variety of cells present in asthmatic lung, including mast cells and eosinophils. Its receptor, c-kit, is broadly expressed on mature mast cells and eosinophils. SCF promotes recruitment of mast cell progenitors into tissues, as well as their local maturation and activation. It also promotes eosinophil survival, maturation and functional activation. SCF enhances IgE-dependent release of mediators from mast cells, including histamine, leukotrienes, cytokines (TNF-alpha, IL-5, GM-CSF) and chemokines (RANTES/CCL5, MCP-1/CCL2, TARC/CCL17 e MDC/CCL22); it is required for IL-4 production in mast cells. SCF, acting in concert with IgE, also upregulates the expression and function of CC chemokine receptors in mast cells. Structural and resident airway cells express increased levels of SCF in the bronchus of asthmatic patients. In a murine model of asthma, allergen exposure increased production of SCF by epithelial cells and alveolar macrophages, which was transient and paralleled by histamine release. SCF induced long-lived airway hyperreactivity, which was prevented by local neutralization of SCF, as well as by inhibitors of the production or activity of cysteinyl-leukotrienes. Together, these observations suggest that SCF has an important role in asthma. PMID- 14561151 TI - Interleukin-5, eosinophilic diseases and therapeutic intervention. AB - Interleukin 5 (IL-5) is the key cytokine in an eosinophil's life span: it supports eosinophilopoiesis and eosinophil differentiation, contributes to eosinophil migration, tissue localisation and function, and prevents eosinophil apoptosis. Given the likely role of eosinophils in chronic inflammatory diseases, a lot of research over the past decade was aimed at antagonising IL-5 function. It appears from recent studies that, although this can easily be achieved in vitro, blocking IL-5 function in vivo is much more difficult than originally anticipated. Here, we review the current status of IL-5 and IL-5 receptor research, with emphasis on strategies to interfere with IL-5 function. PMID- 14561152 TI - The effects of allergen and anti-allergic drugs on murine hemopoietic cells: moving targets, unusual mechanisms, and changing paradigms. AB - We used a variety of techniques to evaluate the effects of airway allergen exposure in mice on the responses of hemopoietic cells to cytokines and drugs in vitro and in vivo. Initial studies have shown that allergen exposure of sensitized mice leads to release of circulating mediators, that induce rapid upregulation of bone-marrow responses to IL-5 and GM-CSF. This may be related to glucocorticoids, because exogenous dexamethasone has similar effects on cultured murine bone-marrow, and because stress-induced glucocorticoids, in naive or sensitized mice, have effects indistinguishable from those of allergen challenge in sensitized animals. Upregulation of eosinophil production is associated with an increased expression of alpha4 integrins, which may contribute to retention of these cells in the bone-marrow. Glucocorticoids regulate the adhesiveness, maturation and survival of eosinophils in murine bone-marrow culture, partly by counteracting the actions of Prostaglandin E2 and possibly other prostanoids. Allergen exposure of sensitized mice leads to accumulation of hemopoietic progenitors in the lungs, which differ from those in bone-marrow in growth properties and sensitivity to glucocorticoids. Lung transplantation has been used to demonstrate that the lung acts as a source of endocrine factors that promote hemopoietic cell accumulation, independently of damage caused by local allergic inflammation. PMID- 14561153 TI - Regulation of granulocyte apoptosis by hemopoietic growth factors, cytokines and drugs: potential relevance to allergic inflammation. AB - It has become apparent that the resolution of inflammation depends on the removal of unwanted inflammatory cells, a process governed by physiological apoptosis and non-inflammatory clearance of apoptotic cells. Granulocytes are central to many of the pathophysiological consequences of uncontrolled inflammatory reactions. Hemopoietic factors and cytokines play a critical role in regulating the longevity of these cells in vitro and in vivo. Here we review the progress that has been made in the understanding of granulocyte apoptosis and the implications for immunotherapy and pharmacological strategies in the treatment of allergic inflammatory diseases for therapeutic gain. PMID- 14561154 TI - CpG oligodeoxynucleotides: a novel therapeutic approach for atopic disorders. AB - Atopic disorders such as allergic rhinitis, asthma and atopic dermatitis are associated with skewing of immune responses towards a TH2 phenotype, resulting in eosinophilic inflammation. TH2 cytokines promote eosinophil growth, migration and activation, mast cell differentiation, and IgE production, and are candidate mediators of pathologic abnormalities in asthma and other atopic diseases. There has been a significant increase in the prevalence of allergic disorders over the past several decades. Recent epidemiological studies suggest that reduced early life exposure to strong TH1 stimuli in industrialized counties has skewed the TH1/TH2 balance towards TH2 responses. Improved hygiene, vaccination, and use of antibiotics may contribute to this imbalance. In the last half of the twentieth century we have seen the use of multiple agents to treat atopic disorders, ranging from antihistamines, steroids and leukotriene modifiers to anti-IgE antibodies. All these agents can block symptoms but do not significantly modify the course of the disease. Recent attempts to restore TH1/TH2 balance by blocking TH2 cytokines or inducing TH1 cytokines, have not only failed to alter the outcome of atopic diseases but, in some cases, have caused significant adverse effects. An alternate method of suppressing TH2 responses takes advantage of the innate immune response to bacterial DNA. Oligodeoxynucleotides (ODN) containing sequence motifs centered on unmethylated CG dinucleotides (CpG ODN) resemble bacterial DNA, and like bacterial DNA are immunostimulatory; we and others have shown that CpG ODN can suppress TH2-mediated atopic inflammation without requiring the induction of TH1-type cytokines. These agents may represent a novel therapeutic approach toward restoring immune tolerance in atopic individuals. PMID- 14561155 TI - Emerging features in the regulation of MMP-9 gene expression for the development of novel molecular targets and therapeutic strategies. AB - Matrix metalloproteinase 9 (MMP-9; gelatinase B) belongs to the subfamily of MMPs that play an important role in tissue remodelling in normal and pathological inflammatory processes. MMP-9 is a major secretion product of macrophages and a component of cytoplasmic granules of neutrophils. The enzyme is also secreted by lymphocytes and stromal cells upon stimulation by inflammatory cytokines, or upon delivery of bi-directional activation signals following integrin-mediated cell cell or cell-extracellular matrix (ECM) contacts. Since the integrity of the tissue architecture is closely dependent of the delicate balance between MMPs and their inhibitors, excessive production of MMP-9 is linked to tissue damage and degenerative inflammatory disorders. As a consequence, regulation of gene transcription and tissue-specific expression of MMP-9 in normal and diseased states are being actively investigated to pave the way for new therapeutic targets. The objective of this article is to provide an overview of recent developments in the field of mmp-9 gene expression in different cell types, from the triggering of cell-surface receptors, to the activation of cytoplasmic mediators and transcription factors responsible for the activation of MMP-9 promoter. We will then focus on emerging evidence showing that transcription of mmp-9 gene can be controlled by epigenetic mechanisms. The usefulness of targeting the signalling pathways regulating MMP-9 expression for the treatment of inflammatory disorders and other indications will be discussed in light of these findings. PMID- 14561156 TI - Effects of chrisotherapeutic gold compounds on prostaglandin E2 production. AB - The mechanism of action of anti-rheumatic gold compounds on 12-O tetradecanoylphorbol 13-acetate (TPA)-induced prostaglandin E(2) (PGE(2)) production in rat peritoneal macrophages were examined. Auranofin (AF) at 3-10 muM inhibited TPA-induced PGE(2) production in a concentration-dependent manner. In the pharmacological experiments, prostaglandin G/H synthase (PGHS)-2-dependent PGE(2) production was inhibited by 10 muM of AF. The enzyme activities of both PGHS-1 and PGHS-2 were not affected by the 10 muM AF. Other gold compounds, aurothioglucose (ATG) and aurothiomalate (ATM) did not inhibit PGE2 production at 10 muM. AF decreased the PGHS-2 protein content, but had no effect on the PGHS-1 protein content. AF at 3-10 muM decreased the PGHS-2 messenger RNA (mRNA) level by RT-PCR determination. Then, the effect of AF on nuclear factor kappa B (NF kappaB), one of the transcription factors known to regulate transcription of a group of proinflammatory proteins, was determined. AF at 1-10 muM inhibited nuclear translocation of NF-kappaB in a concentration-dependent manner. ATG and ATM at 10 muM did not inhibit NF-kappaB nuclear translocation, but with 20 h preincubation, ATG and ATM inhibited PGE(2) production and NF-kappaB nuclear translocation. AF, ATG, and ATM did not affect the binding of NF-kappaB to its specific DNA. These observations may suggest that the effects of gold compounds on the inhibition of NF-kappaB nuclear translocation plays one of the major role in its anti-inflammatory effects in rat peritoneal macrophages. PMID- 14561157 TI - Epithelial cell-derived antibacterial peptides human beta-defensins and cathelicidin: multifunctional activities on mast cells. AB - Antibacterial peptides function as effectors for defense in innate immunity. In mammals, they are implicated in the barrier protection of epithelia where their expression can be induced during infection and inflammation. Over a dozen of antibacterial peptides have been identified in humans. Among these, defensins and cathelicidins have been well characterized. Two types of defensins (alpha- and beta-defensins) are recognized based on the presence of their conserved six cysteine residues, whereas cathelicidins are characterized by a homologous cathelin sequence in the pro-region and a variable antibacterial C-terminal sequence. Human beta-defensins and cathelicidin hCAP18/LL-37 are mainly expressed in epithelial tissues where mast cells are present. Here we review the structure of human beta-defensins and cathelicidin, and describe their multiple activities on mast cells to induce chemotaxis, degranulation and prostaglandin D(2) production, acting through receptors coupled to G-protein-phospholipase C pathway. Thus, in addition to their bactericidal activities, epithelial cell derived antibacterial peptides may modulate the inflammatory responses by recruiting mast cells to inflammation foci and inducing the degranulation as well as prostaglandin production from this cell population. PMID- 14561158 TI - Mechanisms leading to autoantibody production: link between inflammation and autoimmunity. AB - Several lines of evidence have suggested that autoreactive antibodies (Abs) may be triggered by some endogenous antigen (Ag) and undergo affinity maturation toward the target through somatic mutation coupled with isotype switching. To understand the mechanism leading to pathogenic auto-Ab production, it is of great importance to analyze endogenous Ags that can break immunologic tolerance and reveal how the triggered auto-Abs evolve to acquire pathogenicity. As for the first issue, chemically modified self-proteins that are frequently found in inflamed tissues are potential candidates. These post-translational modifications might give rise to the generation of neoepitopes to which T- and B- lymphocytes are not tolerated. In the second part, it is discussed how auto-Abs thus triggered undergo affinity maturation toward target auto-Ags. Recently, not only immature B cells in the bone marrow, but also a population of peripheral B cells have been shown to undergo secondary V(D)J recombination of immunoglobulin genes, thereby changing the Ag-specificity. This process is termed receptor revision, which, along with somatic mutation, is considered to contribute to the formation of high-affinity Abs. Receptor revision and somatic mutation may generate auto Abs if the deletion mechanism of autoreactive clones is impaired. B cells that had undergone receptor revision have been isolated from ectopic germinal centers in inflamed tissues in patients with rheumatoid arthritis. Investigations on the link between inflammation and autoimmunity will provide new aspects for therapeutic targets in the treatment of autoimmune diseases. PMID- 14561159 TI - Inflammatory mechanisms in myocardial infarction. AB - Myocardial infarction is associated with an inflammatory response, ultimately leading to healing and scar formation. Reperfused myocardial infarcts exhibit an enhanced inflammatory reaction, and are associated with improved cardiac repair and patient survival. This review summarizes our current knowledge of the inflammatory mechanisms mediating injury and repair following myocardial ischemia and reperfusion. Myocardial necrosis is associated with complement activation and free radical generation, triggering a cytokine cascade and chemokine upregulation. Interleukin (IL)-8 and C5a are released in the ischemic myocardium, and may have a crucial role in neutrophil recruitment. Extravasated neutrophils may induce potent cytotoxic effects through the release of proteolytic enzymes and the adhesion with Intercellular Adhesion Molecule (ICAM)-1 expressing cardiomyocytes. However, despite these potentially injurious effects, the post reperfusion inflammatory response may significantly enhance healing. Monocyte Chemoattractant Protein (MCP)-1 is induced in the infarcted area and may regulate mononuclear cell recruitment. Accumulation of monocyte-derived macrophages, and mast cells may increase expression of growth factors inducing angiogenesis and fibroblast accumulation in the infarct. In addition, expression of cytokines inhibiting the inflammatory response, such as Interleukin (IL)-10 may suppress injury. Matrix Metalloproteinases (MMPs) and their inhibitors regulate extracellular matrix deposition and play an important role in mediating ventricular remodeling. Inflammatory mediators may induce recruitment of blood derived primitive stem cells in the healing infarct, which may differentiate into endothelial cells and even lead to limited myocardial regeneration. Understanding the cellular and molecular steps involved in regulating infarct healing may lead to specific interventions aimed at optimizing cardiac repair. PMID- 14561161 TI - Cytokine network in inflammatory bowel disease. AB - Disease states, such as those characterized by inflammation of the gastrointestinal tract (Crohn's disease and ulcerative colitis), are due to a host of factors that act in concert to produce pathologic change. The immunologic factors that mediate the development of such mucosal inflammation have been at the forefront of IBD research. Recently, a better understanding of the mechanisms involved in mucosal homeostasis and the occurrence of inflammatory bowel disease (IBD) has been achieved with the advent of animal models of mucosal inflammation. This review discusses these models and the insights they provide into the pathogenesis and regulation of IBD. PMID- 14561160 TI - Inflammatory cytokines and cardiovascular disease. AB - The designation of atherosclerosis as a chronic inflammatory process represents an interesting paradigmatic shift for cardiologists. The plasma concentrations of interleukin-6 and its hepatic byproduct, C-reactive protein, may reflect the intensity of occult plaque inflammation and the vulnerability to rupture. Monocyte chemoattractant protein-1 and interleukin-8 play a crucial role in initiating atherosclerosis by recruiting monocytes/macrophages to the vessel wall, which promotes atherosclerotic lesions and plaque vulnerability. In addition, circulating levels of these proinflammatory cytokines increase in patients with acute myocardial infarction and unstable angina, but not in those with stable angina. Also, the plasma concentrations of these cytokines increase after percutaneous coronary intervention, causing late restenosis after the procedure. Angiotensin II and other atherogenic factors induce these cytokines in the cardiovascular tissues through the activation of transcription factors, such as nuclear factor-kappaB or peroxisome proliferator-activated receptors. Conversely, HMG-CoA reductase inhibitors (statins) can potently inhibit these proinflammatory factors in the vessels. A small GTP-binding protein, Rho, may be a key molecule to explain the anti-inflammatory effects of statins. Interleukin 10 also exerts anti-inflammatory effects on the cardiovascular tissues, possibly by deactivating proinflammatory cytokines and inducible nitric oxide synthase. Gene therapy using interleukin-10 may be a promising means for untreatable or complicated cases of cardiovascular diseases. Thus, therapeutic modulations of these inflammatory cytokines may be useful in the prevention of atherosclerosis and future cardiovascular events. PMID- 14561162 TI - Recent indications and methods of surgery for inflammatory bowel disease. AB - Inflammatory bowel disease, notably ulcerative colitis (UC) or Crohn disease (CD), is basically benign, but sometimes develops into serious or fatal cancer. While the primary therapies are medical, such as pharmacotherapy and dietetic modification, intractable, serious, and cancerous cases can require surgical intervention. Surgery represents only one of the treatment options, but prediction of whether UC and CD are likely to progress to serious conditions and determination of when to undertake surgery is essential. Various surgical procedures have been developed over time, and the postoperative results are now generally good. Regarding laparoscopic surgeries, relatively few cases have been accumulated, and addressing the indications and limitations is premature at this point. However, this procedure is likely to fulfill a central role in surgical treatment strategies and represent a major benefit to patients. This paper discusses surgical treatment indications and methods for UC and CD, and explains the practical aspects of laparoscopic surgery, which has made remarkable progress in recent years, for such cases. PMID- 14561164 TI - IL-6 and Crohn's disease. AB - Interleukin-6 (IL-6) is a pleiotropic cytokine with central roles in immune regulation, inflammation, hematopoiesis, and oncogenesis. Its biological activities are shared by IL-6-family of cytokines such as leukemia inhibitory factor and oncostatin M. When IL-6 binds to IL-6R, the IL-6/IL-6R complex then associates with gp130, the common signal transducer of cytokines related to IL-6. IL-6R does not have to be expressed on the cell surface for IL-6 signaling because soluble form of IL-6R (sIL-6R) can bind to IL-6 and function through gp130. Increased levels of IL-6 and sIL-6R have been demonstrated in both serum and intestinal tissues of the patients with active Crohn's disease. In animal model studies, anti-IL-6R monoclonal antibody (mAb) successfully prevented intestinal inflammation and systemic wasting disease by suppressing adhesion molecule expression by vascular endothelium. It also reduced colonic expression of tumor necrosis factor alpha, IL-1beta, and interferon gamma mRNA without affecting the production of transforming growth factor beta, IL-10, and IL-4. Moreover, the treatment displayed therapeutic efficacy against established colitis through the induction of lamina propria T-cell apoptosis. These results strongly suggest that specific targeting of IL-6/sIL-6R pathway will be a promising new approach for the treatment of Crohn's disease, and the clinical trial of humanized anti-IL-6R mAb has been carried out. PMID- 14561163 TI - Therapeutic approaches to chronic intestinal inflammation by specific targeting of mucosal IL-7/IL-7R signal pathway. AB - Inflammatory bowel disease is thought to result from inappropriate activation of mucosal immune responses. Intestinal epithelial cells produce interleukin (IL)-7 that serves as a regulatory factor for IL-7 receptor (IL-7R)(+) mucosal lymphocytes. The pivotal role of mucosal IL-7/IL-7R dependent signals in the activation of mucosal immune responses that lead to the development of chronic intestinal inflammation are demonstrated. Therapeutic approaches targeting IL 7/IL-7R signal pathway may be feasible in the treatment of inflammatory bowel disease. PMID- 14561165 TI - Macrophage-derived IL-18 targeting for the treatment of Crohn's disease. AB - Crohn's disease is an inflammatory bowel disease associated with several changes in the immune system, including an increased number of infiltrating macrophages. These macrophages release a variety of proinflammatory cytokines, such as tumor necrosis factor-alpha (TNF-alpha, macrophage infiltrating factor (MIF), interleukin (IL)-6, IL-12 and IL-18, which are critically involved in the onset and the development of Crohn's disease. We here focus on the role of macrophages, especially macrophage-derived IL-18 in both patients with Crohn's disease and a murine model of Crohn's disease. PMID- 14561166 TI - Multiparticulate systems in the treatment of inflammatory bowel disease. AB - This article shall give an overview on drug delivery systems under development for new therapeutic strategies in the treatment of inflammatory bowel disease. Local delivery of drugs in the colon is mainly aimed to improve efficacy to side effect profiles. The various features of the different approaches allowing drug delivery to the inflamed colon are discussed including the main physiological and pathophysiological problems for different systems. Developments for colonic delivery by oral administration always tried to adapt the carriers to the physiological requirements which shall further increase the therapeutic efficiency and improve patient compliance. The newer carriers described here shall allow to exclude certain variations by physiological factors like local pH, transit throughout the gastrointestinal tract, the potential role of gut microflora, and drug dissolution in the diseased large intestine, which always have been impeding conventional carrier systems for this treatment. Therefore, especially new strategies for such treatment will be presented including liposomal formulations, nanoparticles, bacterial cytokine expression and viral gene therapy approaches. Effective and selective delivery even of an otherwise non-specifically acting drug could provide new therapeutic pathways in the treatment of inflammatory bowel disease. PMID- 14561167 TI - Probiotics and immune regulation of inflammatory bowel diseases. AB - Intestinal microflora play an important role in the pathogenesis of inflammatory bowel diseases (IBD). Certain probiotic bacteria provide beneficial effects for human health and intervention of IBD. Possible mechanisms underlying these effects are diverse and include many aspects of the interaction of the host with its commensal microflora, with immunological and non-immunological effects. This review paper will focus on the recent progress in the use of probiotics in the treatment of IBD, and discuss the potential immunological mechanisms underlying their effects, such as the modulation of mucosal T cell, B cell, epithelial cell, dendrtic cell, macrophage, nature killer cell, antibody, and cytokine responses. PMID- 14561168 TI - Macrophages in inflammatory bowel disease. AB - Blood monocytes which differentiate into tissue macrophages, are unique in that they can not only initiate immune responses but can also be effector cells which contribute to the resolution of these responses. There is no single activation phenotype, and macrophages can be induced to differentiate into cells that either exacerbate or inhibit acute inflammation. Similarly, these cells can promote, deviate or suppress adaptive immune responses. This review focuses on the mechanisms that have been implicated in the recruitment, activation and differentiation of inflammatory monocytes/macrophages in chronic inflammatory bowel diseases, i.e. ulcerative colitis and Crohn's disease. These mechanisms might provide attractive targets for novel therapies. PMID- 14561169 TI - Role of Fas-ligand induced apoptosis in pulmonary inflammation and injury. AB - In the lung, inflammation followed by the loss of epithelial cell precursors beyond a safeguard threshold, leads to increased mesenchymal repair and autonomous fibrosis. Fas-Fas ligand induced apoptosis promotes IL-1beta secretion, neutrophil extravasation, and loss of epithelial cells. In models of lung disease, inflammation and fibrosis can be controlled by interfering with either Fas-Fas ligand interaction, or with downstream caspase activation. These results suggest that the Fas-Fas ligand pathway is a target for the design of new therapeutic strategies for lung diseases. PMID- 14561170 TI - Regulation of eosinophil migration and Th2 cell function by IL-5 and eotaxin. AB - Asthma is characterized by elevated production of IgE, Th2 cytokines, chemokines, mucus hypersecretion, globlet cell metaplasia/hyperplasia, airway obstruction, eosinophilia and enhanced bronchial hyperresponsiveness. These hallmark features of asthma have all been linked to the effector functions of Th2 cytokines (e.g., interleukin-(IL)-4,5,9,10, and 13) in clinical and experimental investigations. This article will detail some of the pathogenic effects regulated by IL-13, IL-5 and the eotaxin subfamily of chemokines to regulate certain aspects of allergic disease. In particular, the potency of IL-13 in inducing enhanced bronchial responsiveness to spasmogenic stimuli and mucus hypersecretion suggests a key role of this molecule in the induction of airways obstruction. Recent studies also indicate that IL-5 and eotaxin, through eosinophils, may regulate Th2 cell function and IL-13 production from this lymphocyte. Therefore, IL-5 and IL-13 signaling systems are not necessarily mutually exclusive effector mechanisms, but may also be integrated through eosinophils to regulate certain aspects of allergic diseases. Blocking IL-13, or pathways that may promote IL-13-associated allergic lung responses (IL-5 and eotaxin) could provide an important strategy to improve the specificity of asthma therapy. PMID- 14561171 TI - Respiratory tolerance in the protection against asthma. AB - Understanding the pathways involved in the induction and maintenance of respiratory tolerance to airborne allergens is important in designing new therapies for asthma and other allergic diseases that not only control disease symptoms, but also change or potentially cure the disease. Respiratory tolerance, and mucosal immunity are maintained by a complex system of defense mechanisms. Most of the inhaled environmental load is eliminated by exclusion mechanisms, which include physical barriers, such as mucus, and cilia as well as a variety of mediators with anti-microbial and immunomodulatory properties. Blanket immunosuppression is provided by alveolar macrophages, which inhibit antigen presentation and T cell responses, in addition to their role in pathogen elimination. Furthermore, there is antigen specific unresponsiveness or tolerance. This tolerance is mediated by lung dendritic cells producing IL-10, which induce the development of CD4+ T regulatory cells. The development of respiratory tolerance also depends on co-stimulation (CD86, and the ICOS-ICOSL pathway). Although exposure of the respiratory mucosa to some pathogenic agents (especially virus, and endotoxin) is associated with asthma exacerbations, microbial exposure may also promote mucosal tolerance and protection against the development of allergic diseases, but the mechanisms involved are not very well understood. Mucosal-based immunotherapy has been already used as an alternative form of allergen delivery in immunotherapy, the only available treatment that is able to reverse established allergic disease. Strategies to further improve mucosal immunotherapy include the use of modified allergen derived peptides, and adjuvants like CpG motifs. PMID- 14561172 TI - Unconventional strategies for the suppression of allergic asthma. AB - Allergic asthma results from an intrapulmonary allergen-driven Th2 response and is characterized by intermittent airway obstruction, airway hyperreactivity, and airway inflammation. An inverse association between allergic asthma and microbial infections has been observed. And this observation constitutes the base of the hygiene hypothesis. Here we discuss the hygiene hypothesis with emphasis on regulatory cells. We review the evidence for the emergence of regulatory cells, such as CD4(+)CD25(+) T cells during infection or during induction of tolerance by mucosal antigen administration. The review focuses also on the emergence of activated CD8(+) T cells and macrophages, induced by infections or microbial products, which also can result in the suppression of asthma. The underlying mechanisms by which regulatory immune cells suppress asthma may represent novel unconventional strategies controlling asthma. PMID- 14561173 TI - Potential use of drugs that target neural-immune pathways in the treatment of rheumatoid arthritis and other autoimmune diseases. AB - Many autoimmune disorders share two common features, dysregulation of the immune system and stress pathways. Two stress pathways, the hypothalamic-pituitary adrenal (HPA) axis and the sympathetic nervous system (SNS), regulate immune system responses, through release of corticosteroids and norepinephrine (NE), respectively. These neuromediators act on immune cells via specific receptors on their surface to modulate the production of key regulatory cytokines. Glucocorticoids modulate immune responses by glucocorticoid binding to cytoplasmic glucocorticoid receptors within target cells. NE regulates immune responses through interaction with plasma membrane beta- or alpha-adrenergic receptors (AR). Both NE and glucocorticoids promote humoral immunity by altering macrophages and T cell cytokine production after an antigen challenge. Glucocorticoids and NE do this by inhibiting interleukin (IL)-12, and interferon (IFN)-gamma, which drives cell-mediated immunity. Additionally, catecholamines drive humoral immunity by stimulating macrophage IL-10 production. These catecholamine effects are mediated largely via beta(2)-AR activation. Both glucocorticoids and NE inhibit inflammation. However, under some circumstances NE promotes inflammation through interaction with macrophage alpha1-AR and subsequent increases in tumor necrosis factor alpha (TNFalpha production. Although macrophages do not normally express alpha(1)-AR, expression of this receptor on macrophages and monocytes occurs in some disease states, including rheumatoid arthritis (RA). Through these mechanisms the HPA axis and the SNS influence the course and progression of RA. Thus, the HPA axis and the SNS are likely to play key roles in the pathology of RA. Furthermore, therapeutic agents targeting the neural pathways that normally regulate immune system homeostasis may prove beneficial for treating RA and other autoimmune diseases. PMID- 14561174 TI - Immunomodulatory treatment strategies for allergic diseases. AB - Over the last decades the prevalence of allergic disorders, such as hayfever and asthma has increased worldwide, mostly in westernised countries where up to 20 % of the population are affected. The "hygiene hypothesis" suggests that modernised lifestyles such as improved housing conditions, altered dietary habits and smaller family sizes may be responsible for the decrease in infectious and the increase in allergic diseases. Childhood atopic diseases, like eczema, food allergies and recurrent wheezy bronchitis represent a considerable health problem and a major socioeconomic burden due to the chronicity of these disorders. In recent years, a better understanding of the immunopathogenesis of allergic diseases has evolved, which has contributed to the development of novel more targeted forms of therapy. Allergen injection immunotherapy is the only treatment in current use with the potential for modifying the course of allergic disease. In order to better target mucosal allergies, new approaches of administering allergen, via the sublingual or intranasal route, are being developed. The use of modified allergens, allergen peptides, DNA immunization and the use of novel adjuvants represent alternatives to conventional immunotherapy with potential for improved efficacy with less side effects. For atopic asthma, novel treatment strategies aim at locally targeting inflamed airways. Nebulized monoclonal blocking antibodies and soluble interleukin receptors against "Th(2)-type" cytokines have been designed. An alternative approach has been the administration of "Th(1) -type" cytokines. Although, immunomodulatory strategies provide a promising outlook for the treatment of allergic patients, more studies are needed in the future to address issues of efficacy, safety and long-term effects of altered immune responses. PMID- 14561175 TI - Enhanced generation of leukotriene B4 from calcium ionophore-stimulated rat peritoneal inflammatory cells: a possible clinical relevance. AB - Leukotrienes (LTs) producing capacity was investigated in calcium ionophore A23187-stimulated peripheral white blood cells and peritoneal inflammatory cells suspension isolated from the same rat. A reverse phase high performance liquid chromatography technique and computerized UV spectroscopy were employed to isolate and quantitate the released LTs namely, LTC(4) and LTB(4). Preincubation of rat peritoneal inflammatory cells at 37 degrees C for 5 min followed by calcium ionophore A23187 stimulation for another 5 min produced significantly elevated amounts of LTB(4) as compared to peripheral white blood cells isolated from the same rat (103+/-12.7 versus 40+/-3.6 pmol/10(7) cells, respectively; mean+/-SEM). Enhanced generation of LTB(4) was associated with production of similar amounts of LTC(4) as compared with LTC(4) produced by peripheral white blood cells (15.2+/-4.2 versus 14.6+/-2 pmol/10(7) cells, respectively). In subsequent experiments, when peritoneal inflammatory cells and white blood cells suspension isolated from the same rats were stimulated with calcium ionophore A23187 (1 micro M) after preincubation with different concentrations of exogenous arachidonic acid (1, 3 and 10 micro M), significantly higher amounts of LTB(4) were produced by the peritoneal inflamed cells while a similar amounts of LTC(4)were produced by both types of cells. Increased LTB(4) formation by rat peritoneal inflammatory cells may prove to be of pathophysiological relevance, since this compound has been described to play an important role in acute inflammatory reaction. PMID- 14561176 TI - CCR3 and CXCR3 as drug targets for allergy: principles and potential. AB - Atopic disorders include a range of conditions such as allergic asthma, rhinitis, -conjunctivitis, -dermatitis, food and drug allergies and anaphylaxis. Induction of T helper (Th)-2 immune response with consequent IgE dependent eosinophil, basophil and mast cell mediated tissue damage is the characteristic feature of allergies. The mechanism underlying this unique and long appreciated feature of allergy is being elucidated at the molecular level with advances in our knowledge of the chemokine system. Thus, chemokines that target CCR3 in concert with Th2 cytokines appear to play a pathogenic role in allergy. In contrast, chemokines that target CXCR3 in concert with Th1 cytokine appear to play a beneficial role. Accordingly, inhibiting CCR3/Th2 pathway using CCR3 antagonists is viewed as a potentially useful strategy for anti-allergy drug development. In contrast, the idea of using CXCR3 agonists to inhibiting allergic response by promoting CXCR3/Th1 pathway faces serious concerns of their potential pro-inflammatory activities in vivo. In this article we have critically evaluated the literature examining the principle and potential of this anti-allergy drug development strategy including a summary of various compounds that are under investigation. PMID- 14561177 TI - Regulation of inducible nitric oxide synthase by cAMP-elevating phospho diesterase inhibitors. AB - Among the numerous genes controlled by cyclic adenosine monophosphate (cAMP)/protein kinase A signalling machinery is the gene encoding the inducible nitric oxide synthase (iNOS), an enzyme catalyzing the synthesis of a highly reactive free radical nitric oxide (NO). While being a major microbicidal and tumoricidal molecule, iNOS-derived NO has also been implicated in tissue destruction, as well as in regulation of inflammatory/immune cell function in various disorders associated with excessive inflammation. A feasible way for cAMP dependent therapeutic control of inflammation, including iNOS-mediated NO synthesis, could involve the administration of drugs that block the enzymatic activity of cAMP-degrading phosphodiesterases (PDE). Indeed, cAMP-elevating PDE inhibitors can influence iNOS activation in different cell types in vitro, and their potent anti-inflammatory effects in experimental disease models and clinical studies were frequently accompanied with profound modulation of NO production. A set of conflicting data has been generated over the years, ranging from strong suppression to marked enhancement of NO release by cAMP-increasing PDE inhibitors, depending on cell-type, iNOS stimuli, and/or the agents used. The present review summarizes the data on iNOS modulation by cAMP-elevating PDE inhibitors and possible mechanisms behind it, speculating on its contribution to the therapeutic effects of these drugs. PMID- 14561178 TI - Eotaxins and CCR3 receptor in inflammatory and allergic skin diseases: therapeutical implications. AB - Cell migration is mediated by a group of chemotactic cytokines called chemokines: low molecular weight molecules that have been shown as important leukocyte chemical attractants to sites of inflammation and infection. Eotaxin-1, also called CCL11, was first described in 1994, as a highly specific eosinophils chemokine. Many cell types including lymphocytes, macrophages, bronchial smooth muscle cells, endothelial cells and eosinophils, are able to produce this chemokine, predominantly after cytokine stimulation, however little is known about its expression in human skin in vivo. Eotaxin-1 also regulates the chemiotaxis and, in some conditions, activation of basophils, mast cells and T lymphocytes. Chemokine receptors are named from their ligand families, thus the CC chemokine eotaxin-1 binds to the CCR3 receptor which is expressed on eosinophis, mast cells, Th2 type lymphocytes and even on keratinocytes. It seems that eotaxin-1 is one of the most important cytokines involved in tissue inflammation playing a central role in the pathogenesis of allergic airway diseases (asthma and rhinitis), in inflammatory bowel disease and gastrointestinal allergic hypersensitivity and recently it has been proposed as a therapeutical target for these conditions. Our group has studied the role of eotaxin-1 in the pathogenesis of two skin conditions: dermatitis herpetiformis and AIDS-associated eosinophilic folliculitis, demonstrating that this chemokine, together with Th2 type cytokines (IL-13 and IL-4) is important in cell recruitment, inflammation and tissue damage; moreover eotaxin has proven to paly an important role in other skin conditions such as, bullous pemphigoid, pemphigoid gestationis, atopic dermatitis and allergic drug reactions Recent advances in the understanding of eotaxin-1-mediated mechanisms of chemotaxis in allergic and inflammatory conditions may predict that therapeutic antagonism is achievable. This paper will focus on the role that eotaxin and its receptor play in the pathogenetical mechanism in a number of dermatologic diseases, some of which, like atopic dermatitis, may benefit from the introduction of novel and more selective therapeutic options. PMID- 14561179 TI - Emerging treatments for autoimmune hepatitis. AB - Prednisone alone or a lower dose of prednisone in combination with azathioprine induces remission and enhances survival in autoimmune hepatitis. Treatment failure, incomplete response, drug-induced side effects, and relapse after drug withdrawal are unsatisfactory outcomes that justify the search for new therapies. Potent new drugs promise greater blanket immunosuppression than current regimens, and insights into the pathogenic mechanisms of the disease make site-specific interventions possible. Cyclosporine and tacrolimus are calcineurin inhibitors that impair the transcription of interleukin 2, reduce the expression of cytokines, and diminish T lymphocyte proliferation. Mycophenolate mofetil antagonizes the synthesis of purines and depletes stores of guanine nucleotides necessary for DNA synthesis and expansion of T cell clones. Controlled clinical trials are warranted to establish the role of these new drugs in the treatment of autoimmune hepatitis. Promising site-specific therapies include peptides that competitively block autoantigen presentation, agents such as cytotoxic T lymphocyte antigen 4 that inhibit the second co-stimulatory signal of immunocyte activation, T cell vaccination, oral tolerance therapy, and cytokine manipulation with monoclonal antibodies and recombinant supplements. Confident animal models of experimental autoimmune hepatitis are necessary to mature these interventions. In conclusion, promising immunosuppressive agents that alter cytokine expression and T lymphocyte proliferation may be of value in the treatment of autoimmune hepatitis. Critical mechanisms of immunocyte activation, cytotoxic T cell expansion, and cytokine modulation are the targets of site-specific interventions. PMID- 14561181 TI - Novel therapeutic targets for acute pancreatitis and associated multiple organ dysfunction syndrome. AB - Acute pancreatitis is a common clinical condition. The exact mechanisms by which diverse etiological factors induce an attack are unclear but once the disease process is initiated, common inflammatory and repair pathways are invoked. Acinar cell injury early in acute pancreatitis leads to a local inflammatory reaction; if marked, this leads to a systemic inflammatory response syndrome (SIRS). An excessive SIRS leads to distant organ damage and multiple organ dysfunction syndrome (MODS). MODS associated with acute pancreatitis is the primary cause of morbidity and mortality in this condition. The systemic effects of acute pancreatitis have many similarities to those of other conditions such as septicemia, severe burns and trauma. Potentially, there is a therapeutic window between symptom onset and the development of distant organ damage in acute pancreatitis, when anti-inflammatory therapy may be of use. Recent studies conducted by us and other investigators have established the critical role played by inflammatory mediators such as TNF-alpha, IL-1beta, IL-6, IL-8, CINC/GRO alpha, MCP-1, PAF, IL-10, CD40L, C5a, ICAM-1, and Substance P in acute pancreatitis and the resultant MODS. It is reasonable to speculate that elucidation of the key mediators in acute pancreatitis coupled with the discovery of specific inhibitors will make it possible to develop a clinically effective anti-inflammatory therapy. PMID- 14561180 TI - The role of TNF and its family members in inflammation and cancer: lessons from gene deletion. AB - Almost two decades ago, tumor necrosis factor (TNF) was identified as a protein produced by the immune system that played a major role in suppression of tumor cell proliferation. Extensive research since then has revealed that TNF is a major mediator of inflammation, viral replication, tumor metastasis, transplant rejection, rheumatoid arthritis, and septic shock. As of today, 18 different members of the TNF superfamily have been identified, and most of them have been found to mediate a wide variety of diseases including cancer, arthritis, bone resorption, allergy, diabetes, atherosclerosis, myocardial infarction, graft versus host disease, and acquired immune deficiency disease. All the cytokines of the TNF superfamily mediate their effects through the activation of the transcription factor NF-kappaB, c-Jun N-terminal kinase, apoptosis, and proliferation. Thus, agents that can either suppress the production of these cytokines or block their action have therapeutic value for a wide variety of diseases. In this review, we have elucidated the signal transduction pathways used by the members of the TNF family and the effects of deletion of genes that mediate the pathways. Our current understanding of the signaling pathways for TNF and other family members could serve as a target for the development of therapeutics. PMID- 14561182 TI - Peptide-based vaccines in the treatment of specific allergy. AB - The efficacy of conventional allergen-specific immunotherapy (SIT) for allergic conditions and venom hypersensitivity is well documented. However it's use is limited due allergic side effects including anaphylaxis and the difficulty of standardising proteins in complex allergenic mixtures. The aim of new therapeutic strategies is to circumvent these limitations and approaches include allergen non specific therapy, such as anti-IgE and anti-cytokine therapy and other allergen specific techniques including the peptide based vaccines (PBV), modified allergens (allergoids) and DNA vaccines. PBV are small linear peptide fragments containing T cell epitopes which are designed to reduce the ability to cross link antigen-specific IgE. Studies in animal models have confirmed proof of principle demonstrating the induction of hyporesponsiveness using high doses of peptides. However, the principle limitation to clinical use of PBV is the polymorphism of HLA class II molecules. There are ongoing clinical studies using peptide-based vaccines for cat, bee and grass allergies--looking at both immunological mechanisms and clinical outcome measures. The mechanisms underlying the efficacy of PBV appear to be similar to those described for classical immunological tolerance. Thus, the peptides may induce anergy due to absence of co-stimulation, activation-induced cell death, a switch from a Th2 to a Th1 cytokine profile, the induction of regulatory T cells or combinations of these mechanisms. Successful immunotherapy, in bee sensitive individuals, is associated with the elaboration of IL-10. Clonal deletion is unlikely as an overall mechanism as there is evidence that the subsequent in vitro response to associated, non-injected, peptides can be suppressed. Mechanistic studies continue to provide insight into the mode of action of whole allergen and peptide-based immunotherapy. Clinical studies designed on the basis of these observations hold the promise of safer vaccines with improved efficacy. Whether this strategy can be used for allergy to complex allergen mixtures such as dust mites will need further evaluation. PMID- 14561183 TI - Critical roles for bradykinin and prostanoids in acute inflammatory reactions: a search using experimental animal models. AB - Searches for chemical mediators of inflammation underlying classical signs of inflammation i.e. heat, redness, swelling, and pain have been performed and various experimental models for evaluation of new agents to manage these inflammatory signs have been developed extensively during the last century. Now, at the beginning of the 21st-century, after great progress in gene technology, the necessity of in vivo animal study is being reconsidered. Therefore, this review introduces and describes findings obtained by the use of various experimental animal models. We have compared the inflammatory characteristics among species using reported animal models such as dye exudation in the skin, paw edema, pleurisy, and writhing reaction; then we have precisely examined mediators involved in these inflammatory reactions. In the process of plasma exudation and pain perception in the earlier phases of acute inflammation, involvement of the kallikrein-kinin system and prostanoids was demonstrated. Precisely, bradykinin, and PGI(2) among the prostanoids, are major mediators for exudation and pain perception of the initial acute phase of inflammation; both mediators collaborate to enhance these effects. PGE(2), perhaps produced by cyclooxygenase-2, was involved in induction of plasma exudation and pain perception in a later phase than the timing of involvement of PGI(2). Precise roles of various prostanoids will hopefully be clarified by the research projects in progress. PMID- 14561184 TI - TNFalpha as therapeutic target: new drugs, more applications. AB - TNFalpha is a crucial cytokine in the establishment and maintenance of inflammation in multiple autoimmune diseases. With the introduction of infliximab and etanercept, two injectable biologic TNFalpha blocking drugs are now available. Both are effective in the treatment of rheumatoid arthritis, reducing clinical inflammation and damage to bones. In addition, infliximab is FDA approved for the treatment of Crohn's disease. More recent controlled trials have shown effectiveness for TNFalpha blockers in psoriasis, psoriatic arthritis, and ankylosing spondylitis. Further trials are underway in diverse inflammatory conditions including including uveitis, sarcoidosis, Behcet's syndrome, and graft versus host disease. Although the safety profile has been generally excellent, the rare development of reactivation tuberculosis, anti double-stranded DNA antibodies, or a demyelination syndrome point out the need for further close follow-up of treated patients. New formulations of recombinant anti-TNFalpha biologics undergoing clinical trials use modifications to reduce antigenicity, increase the half-life, and maintain or extend the efficacy of these agents. Future development of TNFalpha antagonists is turning to small molecule inhibitors. The inhibition of the TNFalpha signaling cascade is under study using blockers of the p38, JNK, and ERK kinases, and by antagonists of transcription factor NF-kappaB activation. The goal of this approach is to develop compounds that are orally available, have increased selectivity compared to generalized blockade of TNFalpha, yet are therapeutically useful for a range of chronic inflammatory diseases. PMID- 14561185 TI - Role of cytokines and oxidative stress in the pathophysiology of acute pancreatitis: therapeutical implications. AB - Severe acute pancreatitis causes a high incidence of mortality due to the systemic inflammatory response syndrome leading to multiple organ failure. At present, there is no treatment against severe acute pancreatitis, other than supportive critical care. The relationship between pancreatic injury and the uncontrolled systemic response is not completely understood. Nevertheless, experimental and clinical evidences have shown that pro-inflammatory cytokines and oxidative stress are critically involved in the development of local and systemic complications associated with severe acute pancreatitis. Serum levels of pro-inflammatory cytokines, such as TNF-alpha and IL-1beta, increase during the course of acute pancreatitis and they appear to be the driving force for the initiation and propagation of the systemic response. Accordingly, pretreatment with an antibody against TNF-alpha or blockade of TNF-alpha production with pentoxifylline ameliorates experimental acute pancreatitis. In addition, serum IL 6 and IL-8 levels appear to be correlated with severity of pancreatic inflammation. The role of oxidative stress in acute pancreatitis has been evidenced indirectly by beneficial effects of antioxidants as well as directly by pancreatic glutathione depletion and increased lipid peroxidation. Furthermore, circulating xanthine oxidase released by the damaged pancreas acts as a source of systemic oxidative stress contributing to lung inflammation. In conclusion, pancreatic injury seems to trigger at least two different pathways, i.e. pro inflammatory cytokines and oxidative stress, both involved in the systemic effects of acute pancreatitis. Elucidation of these mechanisms and their interactions is critical to develop a treatment based on the pathophysiology of acute pancreatitis. PMID- 14561186 TI - Treatment of allergic rhinitis: H1-antihistamines and intranasal steroids. AB - Allergic rhinitis is charterized as an inflammatory disease of the nasal mucosa. In clinical practice, H(1)-antihistamines and topical corticosteroids are most commonly used pharmacological agents for the treatment of allergic rhinitis. The beneficial effects of steroids depend upon their long-term anti-inflammatory effect rather than upon direct receptor antagonism. This is different to H(1) antihistamines, which block both neural and vascular H(1) receptors and have a clinical effect on symptoms such as nasal itching, sneezing, and rhinorrhea. H(1) antihistamines are rapidly absorbed and most of them are metabolized by the hepatic cytochrome P system and begin to reduce nasal symptoms (itching and sneezing) within one hour. Understanding of both the efficacy and the pharmacological properties of these commonly used drugs in the treatment of nasal allergic inflammation and its related nasal symptoms is very important. From a clinical viewpoint, it will provide a useful guideline for an appropriate use of these drugs. PMID- 14561187 TI - Inflammation and Parkinson's disease. AB - Numerous recent findings indicate the possible involvement of an immune mechanism in the pathogenesis of neurodegeneration. The immune reaction could either act as a primary event, generating changes leading to cell death, or could be a secondary response to neuronal injury. In various neurodegenerative disorders such as Alzheimer's, Huntington's or Pick's disease, Down's syndrome, multiple sclerosis and the AIDS-dementia complex, the inflammatory pathomechanism is strongly supported by experimental and clinical studies. Such inflammatory mechanisms have also been postulated in Parkinson's disease (PD). This review summarizes some generalities about inflammation and immune reactions in the context of the brain, and provides clinical, epidemiological and experimental data showing that inflammation and immunity, or even auto-immunity, could be implicated in PD, either in its initial step or in its progression. Different experimental models useful for studying the role(s) of inflammation and (auto)immunity in the neurodegenerative process of the dopaminergic neurons in PD are examined. The major similarities and differences between PD and other neurodegenerative disorders are discussed. PMID- 14561188 TI - Peroxisome proliferator-activated receptors and the control of inflammation. AB - Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors which form a subfamily of the nuclear receptor gene family. This subfamily consists of three isotypes, alpha (NR1C1), gamma (NR1C3), and beta/delta (NRC1C2) with a differential tissue distribution. PPARalpha is expressed primarily in tissues with a high level of fatty acid catabolism such as liver, brown fat, kidney, heart and skeletal muscle. PPARbeta is ubiquitously expressed, and PPARgamma has a restricted pattern of expression, mainly in white and brown adipose tissues, whereas other tissues such as skeletal muscle and heart contain limited amounts. Furthermore, PPARalpha and gamma isotypes are expressed in vascular cells including endothelial and smooth muscle cells and macrophages/foam cells. PPARs are activated by ligands, such as naturally occurring fatty acids, which are activators of all three PPAR isotypes. In addition to fatty acids, several synthetic compounds, such as fibrates and thiazolidinediones, bind and activate PPARalpha and PPARgamma, respectively. In order to be transcriptionally active, PPARs need to heterodimerize with the retinoid-X-receptor (RXR). Upon activation, PPAR-RXR heterodimers bind to DNA specific sequences called peroxisome proliferator-response elements (PPRE) and stimulate transcription of target genes. PPARs play a critical role in lipid and glucose homeostasis, but lately they have been implicated as regulators of inflammatory responses. The first evidence of the involvement of PPARs in the control of inflammation came from the PPARalpha null mice, which showed a prolonged inflammatory response. PPARalpha activation results in the repression of NF-kappaB signaling and inflammatory cytokine production in different cell types. A role for PPARgamma in inflammation has also been reported in monocyte/macrophages, where ligands of this receptor inhibited the activation of macrophages and the production of inflammatory cytokines (TNFalpha, interleukin 6 and 1beta), although part of the anti-inflammatory effects of these ligands seems to be mediated by a mechanism not involving PPARgamma. All these findings suggest a role of PPARs in the control of the inflammatory response with potential therapeutic applications in inflammation-related diseases. PMID- 14561189 TI - Chlamydial heat shock protein 60 and lipopolysaccharide: potential virulence determinants in atherogenesis. AB - Chlamydia pneumoniae infection is associated with atherosclerosis and may be an emerging risk factor in coronary artery disease. C. pneumoniae can infect, multiply within and modulate the function of all atheroma cell types. Specific chlamydial virulence determinants have been identified that permit interaction with host cells and dysregulate cell function. In particular, chlamydial heat shock protein 60 and lipopolysaccharide may modulate cell function to dysregulate lipid metabolism, induce inflammatory cytokine cascades and trigger production of cross-reactive antibodies that initiate and promote atherogenesis. This paper reviews chlamydial heat shock protein 60 and lipopolysaccharide as potential virulence determinants in atherogenesis. PMID- 14561190 TI - The mechanisms of immune-to-brain communication in inflammation as a drug target. AB - There is considerable evidence that the peripheral immune system can signal the brain to elicit a sickness response during infection and inflammation. The induction of the sickness response involves the expression of proinflammatory cytokines such as interleukin (IL)-1beta, tumor necrosis factor-alpha (TNF alpha), and IL-6, both in the periphery and in the brain. The mechanisms by which peripheral cytokines can affect brain function have been the subject of much debate. The precise mechanisms by which cytokines signal the central nervous system (CNS) are unknown, but possibilities include: 1) the direct entry of cytokine into the brain across the blood-brain barrier by a saturable transport mechanism: 2) the interaction of cytokine with circumventricular organs such as the orgnum vasculosum of the lamina terminalis [OVLT] and area postrema, which lack the blood-brain barrier; and 3) activation of afferent neurons of the vagus nerve. Increasing evidence has suggested that the afferent vagus nerve is an important pathway for immune-to-brain communication. However, there are inconsistent findings for the involvement of the afferent vagus nerve in the mediation of transmitting inflammatory signals to the brain. Thus, we describe here the functional relevance of the vagal afferent nerve in mediating these effects. An understanding of the mechanisms involved in immune-to-brain communication should permit us to create new drugs as therapeutic targets to decrease sickness or promote recovery. This review focuses on recent discoveries of the multipathway mechanisms for the induction of sickness behavior mediated through neuroimmune interactions in the CNS. PMID- 14561191 TI - IL-4 and IL-13: their pathological roles in allergic diseases and their potential in developing new therapies. AB - The incidence of allergic diseases has dramatically increased in recent decades, and it is socially and medically important to establish more useful strategies to overcome allergic disorders. Various kinds of drugs are utilized for allergic patients; however, some cases are unresponsive to these drugs and in others there are undesired adverse effects. On the other hand, a substantial body of evidence has accumulated pointing to the pivotal role of Th2-cytokines, interleukin (IL) 4, and IL-13, in the pathogenesis of bronchial asthma. The evidence is categorized as (1) expression of these cytokines in the bronchial lesions, (2) genetic association of the signaling molecules of these cytokines, (3) analyses of mouse models. In addition, the molecular mechanism of the signal transduction of these cytokines has also been well characterized. Based on such information, IL-4 and IL-13 have emerged as promising means of improving allergic states, and several IL-4/IL-13 antagonists have been developed, among which soluble IL-4 receptor is now in human trials. Identifying the structure of the IL-13 variant and of the IL-4/IL-13-inducing genes would be of great use. It is expected that in the near future, several drugs will emerge based on these strategies, which will give us wider choice in treating patients, depending on the pathogenesis of the diseases. PMID- 14561192 TI - The use of leukotriene modifying drugs in asthma and other respiratory diseases. AB - Only rarely in modern medicine is an entirely new class of drug developed. Recently, a number of drugs that act as leukotriene modifiers (LTM's) have been licensed for use in the treatment of asthma. Airway obstruction in asthma has two key components--bronchoconstriction of airway smooth muscle and airway inflammation. Although a number of mediators are involved in this process, it has been demonstrated that leukotrienes can precipitate both. Leukotrienes are formed in eosinophils, mast cells and neutrophils. LTM's have been shown to attenuate bronchial hyper-reactivity and reduce chemotaxis of inflammatory cells in the asthmatic airway. This article reviews the data from clinical trials of LTM's, discusses their role in asthma therapy and postulates on use in other common respiratory diseases. PMID- 14561193 TI - A role for leptin in the systemic inflammatory response syndrome (SIRS) and in immune response. AB - Leptin was originally identified as an adipocyte-derived cytokine with a key role in the regulation of the energy balance. Subsequent research has, however, revealed that leptin's biological action is not restricted to its effects on appetite and food intake, but rather has a much more pleiotropic character. Evidence is now accumulating that it has important functions in reproduction, hematopoiesis, HPA-axis endocrinology and angiogenesis. In this review, we have focused on the effects of leptin in the immune system, which can be found in both the antigen-specific immunity and in the inflammatory effector system. PMID- 14561194 TI - Oxidative stress and gene transcription in asthma and chronic obstructive pulmonary disease: antioxidant therapeutic targets. AB - Inflammatory lung diseases such as asthma and Chronic Obstructive Pulmonary Disease (COPD) are characterised by systemic and local chronic inflammation and oxidative stress. The sources of the increased oxidative stress in patients with asthma and COPD derive from the increased burden of inhaled oxidants, and from the increased amounts of reactive oxygen species (ROS) generated by several inflammatory, immune and structural cells of the airways. Increased levels of ROS produced in the airways are reflected by increased markers of oxidative stress in the airspaces, sputum, breath, lungs and blood in patients with asthma and COPD. ROS, either directly or via the formation of lipid peroxidation products such as acrolein, 4-hydroxy-2-nonenal and F(2)-isoprostanes, may play a role in enhancing the inflammation through the activation of stress kinases (JNK, MAPK, p38, phosphoinositide 3 (PI-3)-kinase/PI-3K-activated serine-threonine kinase Akt) and redox sensitive transcription factors such as NF-kappaB and AP-1. Recent data have also indicated that oxidative stress and pro-inflammatory mediators can alter nuclear histone acetylation/deacetylation allowing access for transcription factor DNA binding leading to enhanced pro-inflammatory gene expression in various lung cells. Furthermore, oxidative stress may alter the balance between gene expression of pro-inflammatory mediators and antioxidant enzymes in favor of inflammatory mediators in the lung. Thus, the presence of oxidative stress may have important consequences for the pathogenesis of asthma and COPD. Identification of genes that predispose to the development of asthma and COPD may identify novel therapeutic targets. Future work is directed to understand the molecular mechanisms of antioxidants on ROS-mediated cell signaling pathways and inhibition of inflammatory response that would provide information for the development of novel antioxidant therapeutic targets in asthma and COPD. Effective wide spectrum antioxidant therapy that has good bioavailability and potency is urgently needed to control the localised oxidative and inflammatory processes that occur in the pathogenesis of asthma and COPD. In addition, development of such novel antioxidant compounds would be therapeutically useful in monitoring the oxidative and inflammatory biomarkers in the progression/severity of asthma and COPD. PMID- 14561195 TI - The glucocorticoid receptor: molecular mechanism and new therapeutic opportunities. AB - Synthetic glucocorticoids are among the most effective anti-inflammatory drugs available. The activity of this drug class is mediated by the glucocorticoid receptor, a nuclear steroid receptor whose endogenous ligand is the adrenal hormone cortisol. Chronic glucocorticoid treatment is accompanied by serious side effects, reflecting the symptoms of cortisol excess seen in Cushing's syndrome patients. During the past 50 years advances to this drug class have been limited largely to reducing systemic exposure through inhaled delivery and increasing glucocorticoid receptor selectivity. However, a safer oral drug for the treatment of conditions such as rheumatoid arthritis, inflammatory bowel disease, and transplant rejection, still represents a major unmet medical need. Over the past 20 years, mechanisms of glucocorticoid receptor action have been elucidated. Before the gene was even cloned, the glucocorticoid receptor was known to be a ligand-induced DNA binding protein. Identification of hormone response elements in the promoters of metabolic target genes in the liver provided a model for its broad activities. It was since revealed that much of its anti-inflammatory activity is not DNA-dependent after all, but instead is the result of a complex set of protein-protein interactions which lead to transcriptional inhibition of pro-inflammatory targets. Thus, a glucocorticoid receptor ligand that dissociates the DNA binding and protein interaction mediated activities is expected to show improved safety. This review will focus on the scientific advances, which impact the development of this hypothesis and will present a survey of current preclinical drug candidates employing this strategy. PMID- 14561196 TI - Upstream signal transduction of NF-kappaB activation. AB - NF-kappaB is a transcription factor governing the expression of genes involved in the immune response, embryo or cell lineage development, cell apoptosis, cell cycle progression, inflammation, and oncogenesis. During the past few years, considerable attention has been paid to the upstream signaling pathways that lead to the activation of NF-kappaB. Many of these signaling molecules can serve as potential pharmaceutical targets for the specific inhibition of NF-kappaB activation leading to interruption of disease processes. How these molecules interact with each other is however, still a debatable issue. Since many of the signal molecules in this pathway relay more than one of the upstream signals to downstream targets, it has been suggested that the transmission of signals involves a network, rather than a linear sequence in the activation of NF-kappaB. Thus, the detailed elucidation of the upstream signaling molecules involved with NF-kappaB activation will be important to the development of pharmaceutical inhibitors that specifically inhibit the activation of NF-kappaB. Such inhibitors would be predicted to have potent anti-inflammatory and/or anti-carcinogenic effects. PMID- 14561197 TI - Current and future therapeutic strategies to target inflammation in stroke. AB - Experimentally and clinically, stroke is followed by both acute and prolonged inflammatory responses characterized by the production of inflammatory cytokines and leukocyte infiltration into the brain. A debate on whether inflammation after stroke is neurotoxic or participates in brain repair remains unresolved. However, the need to pharmacologically control inflammatory amplification has been commonly acknowledged. The principal challenge of devising successful anti inflammatory strategies for stroke is to understand molecular and temporal interplay of inflammatory and cell-death-inducing processes triggered by cerebral ischemia in both parenchymal and vascular brain cells. This article will review a number of experimental and clinically tested approaches to reduce brain inflammation and damage after stroke (e.g., anti-neutrophil, anti-ICAM-1, anti cytokine strategies) and will suggest potential pathways where novel therapeutic targets may emerge, including transcriptional regulators of inflammatory gene expression (e.g., NF-kappaB, proteasome) and signaling pathways (e.g., ICE cascade, MAPK/MKK/ERK cascade) linked to both inflammation and neuronal cell death. Finally, we will discuss applications of functional genomics technologies in the discovery of stroke diagnostics and therapies. PMID- 14561198 TI - Mechanisms of antihistamines and mast cell stabilizers in ocular allergic inflammation. AB - Mast cells play a central role in allergic reactions and inflammation. Successful anti-allergic therapies have typically targeted mast cell mediators, particularly histamine. Antihistaminic compounds interact with the various histamine receptors found on many cells, whereas other compounds such as disodium cromoglycate, are referred to as mast cell stabilizers, as they inhibit degranulation. Some of the most successful compounds developed recently are dual-action, in that they have both anti-histaminic and mast cell stabilizing activities. Recent trends in pharmaceutical intervention, however, have been focused on the secondary effects of mast cell mediators on epithelial cell adhesion molecule expression and mediator release in the process of allergic inflammation. Since, the ocular mucosa is highly exposed to environmental allergens it is commonly involved in allergic reactions and, as such, has been a useful and accessible model in which to test new therapies in vivo. These ocular allergen provocation studies permit analysis of ocular surface cells and evaluation of tear film mediators. Furthermore, techniques to purify conjunctival mast cells have facilitated the study of the effects of mast cell stabilizing compounds on other mast cell mediators, such as cytokines, and the direct effects of mast cell mediators on epithelial cells in vitro. This review will discuss current understanding of how anti-histamines and mast cell stabilizers work, particularly in the context of molecular mechanisms of ocular allergic inflammation. PMID- 14561199 TI - Toll-like receptors and inflammation in the CNS. AB - The innate immune system recognizes "non-self" by employing a set of germline encoded receptors called Toll-like receptors (TLRs), originally characterized in Drosophila. TLRs are involved in the recognition of various microbial-derived molecules, including lipopolysaccharide (LPS), lipoteichoic acid (LTA), and peptidoglycan (PGN), as well as unmethylated bacterial DNA. The TLR-mediated intracellular signaling pathways converge to activate nuclear factor-kappa B (NF kappaB) and c-Jun N-terminal kinases (JNKs), which induce the transcription of a series of cytokine/chemokine genes that are involved in the initiation or regulation of the inflammatory response. It is now known that, like other peripheral organs, the central nervous system (CNS) is also under constitutive immune surveillance by CNS-resident glial cells (microglia and astrocytes) and CNS-infiltrating immune cells. The recent progress in our understanding of TLR functions in the innate immune response sheds new light on how inflammatory immune responses are initiated within the CNS. In this review, we discuss recent studies on TLRs and their ligands, signal transduction pathways activated by TLRs, and the mechanisms through which these various activation events occur. Finally, we discuss how TLRs might play similar important roles in CNS inflammation. PMID- 14561200 TI - Cytokines in dementias. AB - Knowledge regarding putative inflammatory component(s) participating in Alzheimer's disease (AD) and in vascular dementia (VAD) remains scarce. Recently, we have demonstrated the presence of inflammatory components, such as cytokines, in the CSF of demented patients. Although the initial events triggering the neurodegenerative processes in AD versus VAD may be different and thus lead to different neuropathological outcome, they may initiate a similar cascade of cytokine production in response to neuronal injury. The cytokines released in the CNS may in turn, act in a similar manner in both diseases, amplifying certain pathological changes such as amyloidogenesis and amyloid accumulation in the blood vessels, white matter lesions and angiogenesis. This hypothesis is supported by clinical studies demonstrating the presence of white matter infarcts and cerebrovascular pathology in patients with AD as well as the presence of senile plaques in patients with VAD. This review will focus on the production of pro-inflammatory and anti-inflammatory cytokines in dementia, and their putative role for glia cell activation, amyloidogenesis, vascular changes, white matter damage and neurodegeneration. PMID- 14561201 TI - Eotaxin receptor (CCR3) antagonism in asthma and allergic disease. AB - A range of low molecular weight chemicals have been developed to antagonise the eotaxin receptor, cysteine-cysteine chemokine receptor-3 (CCR3), with the aim of selectively inhibiting eosinophil recruitment into tissue sites. However, the results of recent clinical trials with monoclonal antibodies directed against interleukin-5 (IL-5) question the role of eosinophils in mediating the symptoms of asthma and allergic disease. For this reason, the plans for clinical development of certain CCR3 antagonists have been halted. However, eotaxin 1-3 and a variety of other chemokines interact with CCR3; and this receptor is expressed not only on eosinophils but also on basophils, mast cell subpopulations, activated Th2 cells, macrophages, and airway epithelial cells. Hence, CCR3 is closely associated with asthma and allergy and blockade of this receptor may have pronounced beneficial effects in these diseases. We consider the chemical structures of CCR3 antagonist molecules from a range of pharmaceutical companies, and present an early clinical development plan for a hypothetical CCR3 antagonist. CCR3 antagonists are likely to be safe and effective therapies for allergic diseases, and their clinical pharmacology can readily be defined within phase I/II studies in patients with allergy and asthma. PMID- 14561202 TI - Diagnosis, prevention and treatment of aspirin-induced asthma and rhinitis. AB - Bronchial asthma is not a homogenous disease. Several variants of asthma can be distinguished. One of them is aspirin-induced asthma. In this distinct clinical syndrome, aspirin and most other nonsteroidal anti-inflammatory drugs that inhibit cyclooxygenase-1 precipitate rhinitis and asthma attacks. This type of asthma affects 5-10% of adult asthmatics, but remains largely underdiagnosed. The natural history of aspirin-induced asthma (AIA) has been described, based on an extensive pan-European survey. Aspirin provocation tests with improved diagnostic accuracy have been developed, although no in-vitro tests has been found to be of diagnostic value. Recent interest in AIA has been stirred by the finding of alterations in arachidonate metabolic pathways, leading to cysteinyl-leukotriene overproduction. LTC4 synthase is overexpressed in bronchi and its mRNA is upregulated in peripheral blood eosinophils. The gene coding for LTC4 synthase exists in two common alleles, one of which appears to be associated with a severe, steroid-dependent type of asthma. New highly specific COX-2 inhibitors appear to be a safe alternative for patients with aspirin-induced asthma. PMID- 14561203 TI - Leptin as a novel therapeutic target for immune intervention. AB - The recent cloning of the leptin (obese, ob) gene has determined fundamental insight into the understanding of the regulation of food intake, basal metabolism and reproductive function. Leptin, mainly secreted by adipocytes, belongs to the helical cytokine family and its plasma concentrations correlate with fat mass and respond to changes in energy balance. Initially, leptin was considered as an anti obesity hormone, but experimental evidence has also shown pleiotropic effects of this molecule on hematopoiesis, angiogenesis, lymphoid organ homeostasis and T lymphocyte functions. More specifically, leptin links the pro-inflammatory T helper (Th)-1 immune response to the nutritional status and the energy balance. Indeed, decreased leptin concentrations during conditions of food deprivation lead to impaired immune capabilities. This review focuses on the potential therapeutic utilities for agents that manipulate the leptin-adipocyte axis and discusses novel strategies for an immune intervention in pathologic conditions. PMID- 14561204 TI - 5-lipoxygenase: cellular biology and molecular pharmacology. AB - 5-lipoxygenase (5-LO) plays an essential role in the biosynthesis of leukotrienes, proinflammatory mediators which are mainly released from myeloid cells. Whereas LTB(4) is a potent chemotactic and chemokinetic agent for a variety of leukocytes, the cysteinyl-leukotrienes C, D(4) and E(4) cause vascular permeability and smooth muscle contraction. In view of these properties, leukotriene synthesis inhibitors have been hypothesized to possess therapeutic potential for the treatment of asthma, allergic disorders and other inflammatory diseases. Whereas cysteinyl-leukotriene receptor antagonists possess potential for antileukotriene therapy in asthma, results from clinical trials with leukotriene synthesis inhibitors were less encouraging. The expression of 5-LO in mammals is tightly regulated. Enzymatic activity in vitro can be modulated by calcium, ATP, phosphatidylcholine and lipid hydroperoxides; nevertheless activation of cellular 5-LO in response to external stimuli is rather incompletely understood. Intensive research revealed that on cell stimulation, 5 LO redistributes to the nuclear membrane where it colocalizes with 5-lipoxygenase activating protein and cytosolic phospholipase A(2). In addition, various cellular proteins interacting with 5-LO have been identified. It was suggested that enzyme phosphorylation could influence redistribution and cellular activity of 5-LO and that the cellular redox tone regulates 5-LO product formation. This review highlights the determinants of cellular 5-LO activity and summarizes the molecular pharmacology of 5-LO. PMID- 14561205 TI - Oral tolerance in the treatment of rheumatoid arthritis. AB - Oral tolerance (OT) consists of the oral administration of antigens (Ag) that could alter the response of the immune system. This is a form of peripheral immune tolerance in which mature lymphocytes in the peripheral lymphoid tissues are rendered non functional or hyporesponsive by prior oral administration of Ag. It was first described in 1911 in animal models of anaphylaxis. This therapeutic approach requires the orally administration of Ag and the active participation of the gut-associated lymphoid tissue (GALT), a tissue comprising Peyer's patches, intraepitelial cells and villi containing epithelials cells which is a well organized immune network. The mechanisms by which OT is mediated included deletion or anergy and active cellular suppression. The primary factor determining which form of tolerance will be developed after oral administration of Ag is the Ag dosage. Thus, it is thought that low doses of Ag induce the generation of active suppression, via regulatory T cells in the GALT, which then migrate to the systemic immune system. These regulatory T cells produce down regulatory cytokines such as IL4, IL10 and TGFbeta, a Th2 / Th3 cytokine pattern. Conversely, high dose of Ag favors anergy or clonal deletion. The phenomenon in which regulatory cells, as generated by oral tolerization, are primed in an Ag specific manner, but act in the respective microenvironment in a non-Ag specific manner is called bystander suppression. This phenomenon is of particular interest and explained the use of OT in T cell mediated autoimmune diseases such as rheumatoid arthritis (RA), multiple sclerosis (MS) and type I diabetes, some diseases in which the autoAg remains unknown or where there are reactivities to multiple autoAgs. There were several studies demonstrating the effectiveness of orally administered Ag in different animal models of autoimmune diseases, such as experimental allergic encephalomyelitis, collagen induced arthritis, diabetes, but also uveitis, myastenia gravis and transplantation. These mouse or rat models of autoimmune diseases gave the rationale for the therapeutic use of OT in human and this therapeutic approach has been tried in MS and RA, using oral myelin or oral collagen, respectively. In RA, 4 trials of oral type II collagen (CII) in RA have been published. Taken together, these studies suggested that oral CII in RA gave a trend toward clinical improvement, with significance in only 2 studies. Bacterial extract from Escherichia coli containing heat shock proteins has been tried in oral treatment for RA. Two placebo controlled trials and 2 comparative studies gave favorable results for this bacterial extract with no or mild adverse events. Although oral/mucosal tolerance has given successful results in animal models of autoimmune diseases, the enthusiasm for this therapeutic approach in human diseases must be tempered. The discrepancies between the effectiveness of OT in animal models and the results in human trials raise some questions, the identification of the subgroup of patients who might respond to this treatment and the source (or nature) of the administered Ag (homologous versus heterologous), for instance. PMID- 14561206 TI - Endocannabinoid degradation, endotoxic shock and inflammation. AB - Endocannabinoids are an emerging class of lipid mediators, which include amides and esters of long chain polyunsaturated fatty acids. Anandamide (N arachidonoylethanolamine, AEA) and 2-arachidonoylglycerol (2-AG) are the main endogenous agonists of cannabinoid receptors. Endotoxic shock is a potentially lethal failure of multiple organs that can be initiated by the inflammatory agent lipopolysaccharide (LPS), present in the outer membrane of gram-negative bacteria. LPS has been recently shown to stimulate the production of AEA in rat macrophages, and of 2-AG in rat platelets. The mechanism responsible for this effect has not been elucidated. On the other hand, mast cells are multifunctional bone marrow-derived cells found in mucosal and connective tissues and in the nervous system, where they play an essential role in inflammation. As yet, little is known about endogenous modulators and mechanisms of mast cell activation. Here, we review recent literature on the role of endocannabinoids in endotoxic shock and inflammation, and report our recent research on the effects of LPS on the production of AEA and 2-AG in human lymphocytes, and on AEA degradation by a specific AEA membrane transporter (AMT) and an AEA-degrading enzyme (fatty acid amide hydrolase, FAAH). We also report the ability of the HMC-1 human mast cells to degrade AEA through a nitric oxide-sensitive AMT and a FAAH. The role of endocannabinoids in HMC-1 degranulation is discussed as well. Taken together, it can be suggested that human lymphocytes and mast cells take part in regulating the peripheral endocannabinoid system, which can affect some activities of these cells. PMID- 14561207 TI - An update of immunotherapy for specific allergies. AB - Allergic diseases are common, disabling and potentially life threatening. The processes that lead to production of excessive allergen-specific IgE production are highly complex and heterogeneous. While current treatment strategies are limited, recent technological advances have provided a better understanding of underlying disease processes and offered new potential therapeutic targets. Optimal treatment strategies permanently modify underlying inflammatory allergic immune responses (immunotherapy) with long term alleviation of symptoms and minimal side-effects. Although these processes are still not completely understood, methods of modifying allergen recognition by the immune system have already been successful. Here, we review recent developments and future directions in allergen immunotherapy and adjunctive therapies. Specifically, we address the molecular mechanisms of allergen immunotherapy and new techniques including allergen modification, allergen gene vaccination, CpG immunostimulation, and peptide immunotherapy. Other non-allergen specific molecular targets (including receptor, cytokine and IgE targets) which may complement specific immunotherapy are also discussed. Ideally these methods will eventually be replaced by strategies targeting the prevention of allergic responses (immunoprophylaxis). PMID- 14561208 TI - The immunosuppressive activity of proinflammatory cytokines in experimental models: potential for therapeutic intervention in autoimmunity. AB - The role of cytokines in the pathogenesis of T cell-mediated diseases, and in particular autoimmune responses, has been the subject of intense investigation in the past few years. Transgenic strains of mice have been generated, each expressing individual cytokines in organs targeted by autoimmunity. These animal models provide the most advanced tool available for analyzing the relationship between cytokines and T-dependent autoimmune responses. On the one hand, these experiments confirm that the local expression of proinflammatory cytokines is pivotal in initiating and maintaining pathogenic responses to self. On the other hand, and somewhat unexpectedly, these models have also revealed that cytokine factors controlling autoimmunity can act both as potentiating and inhibitory agents, depending upon the site and timing of exposure. As a result, one major concept emerging from different experimental models, including those originally established in our laboratory, is that proinflammatory cytokines may ameliorate autoimmunity. In this review, we analyze the mechanisms whereby cytokines that are considered as proinflammatory may in contrast suppress immune responses to self antigens. Besides emphasizing that the proinflammatory/immunogenic properties of a given cytokine may not be an intrinsic property, we review evidence that the regulation imposed by the cytokine network on autoimmunity is a finely tuned balance between activation and downmodulation of an individual autoreactive T cell repertoire. By emphasizing that factors such as the duration of cytokine exposure and the type of cell population involved strongly influence that balance, we underline the potential therapeutic implications of cytokine mediated modulation of autoimmunity. PMID- 14561209 TI - Inducible nitric oxide synthase--time for reappraisal. AB - Inducible nitric oxide synthase (iNOS) is one of three key enzymes generating nitric oxide (NO) from the amino acid L-arginine. iNOS-derived NO plays an important role in numerous physiological and pathophysiological conditions, e.g. blood pressure regulation, inflammation, infection, and the onset and progression of malignant diseases. iNOS has been conjectured both as a marker and a therapeutic target in these situations. iNOS is a mediator of unspecific host defence, central in the clearance of bacterial, viral, fungal and parasitic infections. However, excess production of NO appears to be linked to tissue damage and organ dysfunction, e.g. the hypotensive and vasoplegic state characteristic for septic shock. However, the use of iNOS-inhibitors in septic patients should be performed carefully with regard to the essential functions and properties of NO in blood pressure/blood flow regulation. Considering iNOS derived NO as a multifactorial transmitter of tumorigenesis and tumor progression, it is tempting to speculate on therapeutical interference with iNOS activity, especially in tumors where metastatic activity, host denfence mechanisms and the level of differentiation seem to be correlated to iNOS expression. It is the aim of this review to provide basic insights into the NOS family of enzymes as well as their regulation. In the second part of the review, we will point out the pivotal roles NOS play in inflammation and neoplastic diseases. PMID- 14561211 TI - Mode of action of long-term low-dose macrolide therapy for chronic sinusitis in the light of neutrophil recruitment. AB - Chronic sinusitis is a common inflammatory upper respiratory tract disease. One of the prominent features of this disease is persistent purulent effusion containing numerous emigrated neutrophils in the paranasal sinuses. Recent advances in sinusitis research have revealed two positive feedback mechanisms that explain the chronic neutrophil accumulation in the sinus. First, interleukin (IL)-1beta secreted by monocytes, macrophages and fibroblasts upregulates the expression of E-selectin and intercellular adhesion molecule-1 (ICAM-1) in vascular endothelial cells, and thereby induces the extravascular transmigration of neutrophils. The emigrated neutrophils then secrete IL-1beta, which amplifies the expression of E-selectin and ICAM-1, resulting in further neutrophil infiltration. Second, chemoattractants including IL-8 in the sinus effusion initiate neutrophil exudation. Emigrated neutrophils then secrete IL-8, which elicits further neutrophil accumulation in the sinus effusion. Long-term low-dose macrolide therapy was first introduced for the treatment of diffuse panbronchiolitis in the 1980's. In the 1990's it was also shown to be an effective treatment for chronic sinusitis. The inhibitory effect of macrolides on neutrophil infiltration in inflammatory sites has been well documented in these diseases. Several lines of evidence indicate that macrolides do not function simply as a bactericide. In vitro studies have demonstrated various effects of macrolides on immunocompetent cells, inflammatory cells and airway epithelial cells. It has been shown that macrolides inhibit the production of IL-8 and IL 1beta and the expression of ICAM-1, suggesting that macrolides block the aforementioned dual positive feedback system of neutrophil recruitment and thereby exert their clinical efficacy in the treatment of chronic sinusitis. The inhibitory effects of macrolides on multiple steps in the process of neutrophil recruitment are presumably mediated by the inhibition of transcription factors such as nuclear factor-kB and activator protein-1. Further investigation of the mode of action of macrolides at the molecular level would lead to the development of safer and more effective drugs for the treatment of chronic sinusitis. In addition, the possible risk of this therapy such as the occurrence of resistant strains have to be carefully surveyed hereafter. PMID- 14561210 TI - Chemokines in allergy. AB - Allergic diseases such as atopic dermatitis, asthma, and allergic rhinitis represent a significant healthcare problem. Understanding these diseases as dysregulated inflammatory responses has led to many new targets for therapeutic intervention. Recent data concerning soluble IL-4 receptor, monoclonal antibodies against IL-5 and an antibody toward IgE have lead to an appreciation of the crucial role played by Th2 subset of CD4(+) T cells and their corresponding cytokines. While these potential drugs are presently in clinical trials and may be valuable therapeutics, orally bioavailable small molecule inhibitors of Th2 cell responses would be desirable for treatment of these chronic diseases. One strategy is to prevent effector cell migration (Th2 cells, mast cells, and eosinophils) via chemokine receptor antagonism with a suitable small molecule. Chemokine receptors are a subset of the seven transmembrane-spanning family, which mediate their effects through interaction with heterotrimeric G-proteins. The ligands are a structurally related set of proteins that are selectively expressed in certain disease settings. Three chemokine receptors CCR3, CCR4, and CCR8 are preferentially expressed by Th2 cells, mast cells and eosinophils and therefore represent therapeutic targets for allergy. This mini-review will focus on new research involving CCR3, CCR4 and CCR8. The cellular distribution of each receptor, the corresponding chemokine ligands, and various validation studies are discussed. Recent drug discovery advances concerning pharmacological tools and small molecule receptor antagonists will also be presented. PMID- 14561212 TI - Arachidonic acid, palmitic acid and glucose are important for the modulation of clonal pancreatic beta-cell insulin secretion, growth and functional integrity. AB - Insulin-resistant states such as obesity can result in an increase in the function and mass of pancreatic beta-cells, so that insulin secretion is up regulated and Type II diabetes does not develop. However, expansion of beta-cell mass is not indefinite and may well decrease with time. Changes in circulating concentrations of nutritional factors, such as fatty acids and/or glucose, may lead to a reduction in beta-cell mass in vivo. Few previous studies have attempted to explore the interplay between glucose, amino acids and fatty acids with respect to beta-cell mass and functional integrity. In the present study, we demonstrate that culture of clonal BRIN-BD11 cells for 24 h with the polyunsaturated fatty acid arachidonic acid (AA) increased beta-cell proliferation and enhanced alanine-stimulated insulin secretion. These effects of AA were associated with significant decreases in the cellular consumption of D glucose and L-alanine as well as decreased rates of production of nitric oxide and ammonia. Conversely 24 h exposure to the saturated fatty acid palmitic acid (PA) was found to decrease beta-cell viability (by increasing apoptosis), increase the intracellular concentration of triacylglycerol (triglyceride), while inhibiting alanine-stimulated insulin secretion. These effects of PA were associated with significant increases in D-glucose and L-glutamine consumption as well as nitric oxide and ammonia production. However, L-alanine consumption was decreased in the presence of PA. The effects of AA, but not PA, were additionally dependent on glucose concentration. These studies indicate that AA may have a critical role in maintaining the appropriate mass and function of islet beta cells by influencing rates of cell proliferation and insulin secretion. This regulatory effect may be compromised by high circulating levels of glucose and/or PA, both of which are elevated in Type II diabetes and may impact upon dysfunctional and apoptotic intracellular events in the beta-cell. PMID- 14561213 TI - Effects of oxidative stress on endothelial function after a high-fat meal. AB - Postprandial lipaemia is known to cause endothelial dysfunction, but its underlying mechanism is still under debate. The present study was undertaken to investigate the effects of postprandial lipaemia on endothelial dysfunction and oxidative stress. We measured plasma glutathione peroxidase (GSH-Px), an antioxidant enzyme, and the urinary excretion of 8-epi-prostaglandin F2alpha (8 PGF2alpha), a free radical-catalysed product from the oxidative modification of arachidonic acid, in 16 healthy subjects (mean age, 30 +/- 5 years) without major coronary risk factors. Plasma high-sensitive C-reactive protein, soluble intercellular cell-adhesion molecule-1 and vascular cell-adhesion molecule-1 were also measured. High-resolution ultrasound was used to assess the flow-mediated vasodilatation (FMD) of the brachial artery. Blood and urine samples were collected before and 2, 4 and 6 h after a standard high-fat meal (3677 J, containing 50 g of fat). Serum triacylglycerol (triglyceride) increased and FMD decreased significantly after a high-fat meal. Plasma GSH-Px significantly decreased from 27.2 +/- 12.3 microg/ml to 25.7 +/- 11.8 microg/ml (P=0.022) 2 h after the meal, and urinary excretion of 8-PGF2alpha significantly increased from 1286 +/- 1401 pg/mg of creatinine to 2197 +/- 1343 pg/mg of creatinine (P=0.014) at 4 h after the meal. However, there were no significant changes in the levels of high-sensitive C-reactive protein and adhesion molecules after a high-fat meal. In conclusion, endothelial dysfunction was observed after consuming a high fat meal and is associated with augmented oxidative stress manifested by the depletion of serum antioxidant enzymes and increased excretion of oxidative modification products. PMID- 14561214 TI - Intermittent altitude exposures reduce acute mountain sickness at 4300 m. AB - Acute mountain sickness (AMS) commonly occurs at altitudes exceeding 2000-2500 m and usually resolves after acclimatization induced by a few days of chronic residence at the same altitude. Increased ventilation and diuresis may contribute to the reduction in AMS with altitude acclimatization. The aim of the present study was to examine the effects of intermittent altitude exposures (IAE), in combination with rest and exercise training, on the incidence and severity of AMS, resting ventilation and 24-h urine volume at 4300 m. Six lowlanders (age, 23 +/- 2 years; body weight, 77 +/- 6 kg; values are means +/- S.E.M.) completed an Environmental Symptoms Questionnaire (ESQ) and Lake Louise AMS Scoring System (LLS), a resting end-tidal partial pressure of CO2 ( PETCO2) test and a 24-h urine volume collection at sea level (SL) and during a 30 h exposure to 4300 m altitude-equivalent (barometric pressure=446 mmHg) once before (PreIAE) and once after (PostIAE) a 3-week period of IAE (4 h.day(-1), 5 days.week(-1), 4300 m). The previously validated factor score, AMS cerebral score, was calculated from the ESQ and the self-report score was calculated from the LLS at 24 h of altitude exposure to assess the incidence and severity of AMS. During each IAE, three subjects cycled for 45-60 min.day(-1) at 60-70% of maximal O2 uptake (VO2 max) and three subjects rested. Cycle training during each IAE did not affect any of the measured variables, so data from all six subjects were combined. The results showed that the incidence of AMS (%), determined from both the ESQ and LLS, increased (P<0.05) from SL (0 +/- 0) to PreIAE (50 +/- 22) at 24 h of altitude exposure and decreased (P<0.05) from PreIAE to PostIAE (0 +/- 0). The severity of AMS (i.e. AMS cerebral symptom and LLS self-report scores) increased (P<0.05) from SL (0.02 +/- 0.02 and 0.17 +/- 0.17 respectively) to PreIAE (0.49 +/- 0.18 and 4.17 +/- 0.94 respectively) at 24 h of altitude exposure, and decreased (P<0.05) from PreIAE to PostIAE (0.03 +/- 0.02 and 0.83 +/- 0.31 respectively). Resting PETCO2 (mmHg) decreased (i.e. increase in ventilation; P<0.05) from SL (38 +/- 1) to PreIAE (32 +/- 1) at 24 h of altitude exposure and decreased further (P<0.05) from PreIAE to PostIAE (28 +/- 1). In addition, 24-h urine volumes were similar at SL, PreIAE and PostIAE. In conclusion, our findings suggest that 3 weeks of IAE provide an effective alternative to chronic altitude residence for increasing resting ventilation and reducing the incidence and severity of AMS. PMID- 14561215 TI - In self-defence: hexokinase promotes voltage-dependent anion channel closure and prevents mitochondria-mediated apoptotic cell death. AB - In tumour cells, elevated levels of mitochondria-bound isoforms of hexokinase (HK I and HK-II) result in the evasion of apoptosis, thereby allowing the cells to continue proliferating. The molecular mechanisms by which bound HK promotes cell survival are not yet fully understood. Our studies relying on the purified mitochondrial outer membrane protein VDAC (voltage-dependent anion channel), isolated mitochondria or cells in culture suggested that the anti-apoptotic activity of HK-I occurs via modulation of the mitochondrial phase of apoptosis. In the present paper, a direct interaction of HK-I with bilayer-reconstituted purified VDAC, inducing channel closure, is demonstrated for the first time. Moreover, HK-I prevented the Ca(2+)-dependent opening of the mitochondrial PTP (permeability transition pore) and release of the pro-apoptotic protein cytochrome c. The effects of HK-I on VDAC activity and PTP opening were prevented by the HK reaction product glucose 6-phosphate, a metabolic intermediate in most biosynthetic pathways. Furthermore, glucose 6-phosphate re-opened both the VDAC and the PTP closed by HK-I. The HK-I-mediated effects on VDAC and PTP were not observed using either yeast HK or HK-I lacking the N-terminal hydrophobic peptide responsible for binding to mitochondria, or in the presence of an antibody specific for the N-terminus of HK-I. Finally, HK-I overexpression in leukaemia derived U-937 or vascular smooth muscle cells protected against staurosporine induced apoptosis, with a decrease of up to 70% in cell death. These results offer insight into the mechanisms by which bound HK promotes tumour cell survival, and suggests that its overexpression not only ensures supplies of energy and phosphometabolites, but also reflects an anti-apoptotic defence mechanism. PMID- 14561216 TI - Decreased expression levels of rat liver glutathione S-transferase A2 and albumin during the acute phase response are mediated by HNF1 (hepatic nuclear factor 1) and IL6DEX-NP. AB - The acute phase response is characterized by positive and negative regulation of many liver proteins including GSTs (glutathione S-transferases) and albumin. The expression of albumin and some GSTs are dependent on HNF1 (hepatic nuclear factor 1). Interleukin 6 plus dexamethasone induce a nuclear protein (IL6DEX-NP) in rat hepatocytes in vitro that binds to a promoter element adjacent to the HNF1 site of rGSTA2 and decreases its expression. We determined how HNF1 and IL6DEX-NP regulate rGSTA2 and albumin expression in rats during the acute phase response after LPS (lipopolysaccharide) treatment. Expression of rGSTA2 and albumin mRNA decreased 3 h after LPS treatment and remained low for 48 h. Transcription rates showed a similar pattern but albumin transcription was less affected. HNF1 and IL6DEX-NP binding to the rGSTA2 promoter was present in control livers but was absent at 3 and 6 h after LPS. By 12 h, HNF1 and IL6DEX-NP binding to the rGSTA2 promoter reappeared and increased to above normal at 48 h. The patterns of HNF1 and IL6DEX-NP binding to the albumin promoter were similar. Affinity of IL6DEX-NP for the albumin promoter was less than that for the rGSTA2 promoter and changes in the transcription rates were consistent with the difference. Early decreases in rGSTA2 and albumin during the acute phase response are due to decreased binding of HNF1. Later persistent decreases in transcriptional rate of rGSTA2 and to a lesser extent albumin are due to increased IL6DEX-NP binding. IL6DEX-NP appears to be an important negative regulator of gene expression in vitro and in vivo. PMID- 14561218 TI - Paenibacillus sp. TS12 glucosylceramidase: kinetic studies of a novel sub-family of family 3 glycosidases and identification of the catalytic residues. AB - GCase (glucosylceramidase) from Paenibacillus sp. TS12, a family 3 glycosidase, hydrolyses the beta-glycosidic linkage of glucosylceramide with retention of anomeric configuration via a two-step, double-displacement mechanism. Two carboxyl residues are essential for catalysis, one functioning as a nucleophile and the other as a general acid/base catalyst. p-nitrophenyl beta-D glucopyranoside [K(m)=0.27+/-0.02 mM and kcat/K(m)=(2.1+/-0.2)x10(6) M(-1) x s( 1)] and 2,4-dinitrophenyl beta-D-glucopyranoside [K(m)=0.16+/-0.02 mM and k(cat)/K(m)=(2.9+/-0.4)x10(6) M(-1) x s(-1)] were used for continuous assay of the enzyme. The dependence of kcat (and kcat/K(m)) on pH revealed a dependence on a group of pK(a)< or =7.8 in the enzyme-substrate complex which must be protonated for catalysis. Incubation of GCase with 2,4-dinitrophenyl 2-deoxy-2 fluoro-beta-D-glucopyranoside caused time-dependent inactivation (K(i)=2.4+/-0.7 mM and k(i)=0.59+/-0.05 min(-1)) due to the accumulation of a trapped glycosyl enzyme intermediate. Electrospray ionization MS analysis of the peptic digest of this complex showed that the enzyme was covalently labelled by the reagent at Asp 223, consistent with its role as nucleophile. A mutant modified at this residue (D223G) showed substantially reduced activity compared with the wild type (>10(4)), but this activity could be partially restored by addition of formate as an external nucleophile. Kinetic analysis of the mutant E411A indicated that Glu 411 serves as the general acid/base catalytic residue since this mutant was pH independent and since considerable GCase activity was restored upon addition of azide to E411A, along with formation of a glycosyl azide product. PMID- 14561217 TI - Localization of dynein light chains 1 and 2 and their pro-apoptotic ligands. AB - The dynein and myosin V motor complexes are multi-protein structures that function to transport molecules and organelles within the cell. DLC (dynein light chain) proteins, found as components of both dynein and myosin V motor complexes, connect the complexes to their cargoes. One of the roles of these motor complexes is to selectively sequester the pro-apoptotic 'BH3-only' (Bcl-2 homology 3-only) proteins, Bim (Bcl-2-interacting mediator of cell death) and Bmf (Bcl-2-modifying factor), and so regulate their cell death-inducing function. In vivo DLC2 is found exclusively as a component of the myosin V motor complex and Bmf binds DLC2 selectively. On the other hand, Bim interacts with DLC1 (LC8), an integral component of the dynein motor complex. The two DLCs share 93% sequence identity yet show unambiguous in vivo specificity for their respective BH3-only ligands. To investigate this specificity the three-dimensional solution structure of DLC2 was elucidated using NMR spectroscopy. In vitro structural and mutagenesis studies show that Bmf and Bim have identical binding characteristics to recombinant DLC2 or DLC1. Thus the selectivity shown by Bmf and Bim for binding DLC1 or DLC2, respectively, does not reside in their DLC-binding domains. Remarkably, mutational analysis of DLC1 and DLC2 indicates that a single surface residue (residue 41) determines the specific localization of DLCs with their respective motor complexes. These results suggest a molecular mechanism for the specific compartmentalization of DLCs and their pro-apoptotic cargoes and implicate other protein(s) in defining the specificity between the cargoes and the DLC proteins. PMID- 14561219 TI - Cystinuria-specific rBAT(R365W) mutation reveals two translocation pathways in the amino acid transporter rBAT-b0,+AT. AB - Apical reabsorption of dibasic amino acids and cystine in kidney is mediated by the heteromeric amino acid antiporter rBAT/b(0,+)AT (system b(0,+)). Mutations in rBAT cause cystinuria type A, whereas mutations in b(0,+)AT cause cystinuria type B. b(0,+)AT is the catalytic subunit, whereas it is believed that rBAT helps the routing of the rBAT/b(0,+)AT heterodimeric complex to the plasma membrane. In the present study, we have functionally characterized the cystinuria-specific R365W (Arg(365)-->Trp) mutation of human rBAT, which in addition to a trafficking defect, alters functional properties of the b(0,+) transporter. In oocytes, where human rBAT interacts with the endogenous b(0,+)AT subunit to form an active transporter, the rBAT(R365W) mutation caused a defect of arginine efflux without altering arginine influx or apparent affinities for intracellular or extracellular arginine. Transport of lysine or leucine remained unaffected. In HeLa cells, functional expression of rBAT(R365W)/b(0,+)AT was observed only at the permissive temperature of 33 degrees C. Under these conditions, the mutated transporter showed 50% reduction of arginine influx and a similar decreased accumulation of dibasic amino acids. Efflux of arginine through the rBAT(R365W)/b(0,+)AT holotransporter was completely abolished. This supports a two-translocation-pathway model for antiporter b(0,+), in which the efflux pathway in the rBAT(R365W)/b(0,+)AT holotransporter is defective for arginine translocation or dissociation. This is the first direct evidence that mutations in rBAT may modify transport properties of system b(0,+). PMID- 14561223 TI - The effects of superior ovarian nerve sectioning on ovulation in the guinea pig. AB - The effects on spontaneous ovulation associated with the unilateral or bilateral sectioning of the superior ovarian nerves (SON) were analyzed in guinea pigs at different time intervals of the estrous cycle. Day 1 of the estrous cycle was defined as the day when the animal presents complete loss of the vaginal membrane (open vagina). Subsequent phases of the cycle were determined by counting the days after Day 1. All animals were autopsied on the fifth day of the estrous cycle after surgery. Sectioning the right, left, or both SONs on day 5 (early luteal phase) resulted in a significant increase in the number of fresh corpora lutea. Ovulation increased significantly when the left SON (L-SON) was sectioned during late follicular phase (day 1) and medium luteal phase (day 8). When surgery was performed on days 1 or 8, neither sectioning the right SON (R-SON) nor sectioning the SON bilaterally had an apparent effect on ovulation rates. Similarly, ovulation rates were not affected when unilateral (right or left) or bilateral sectioning of the SON was performed during late luteal phase two (day 12). Unilateral or bilateral sectioning of the SON performed during the early luteal phase (day 5) was associated with a significant decrease in uterine weight. A comparable effect was observed when the L-SON was sectioned during late follicular phase (day 1), or medium luteal phase (day 8). No effects on uterine weight were observed when unilateral or bilateral sectioning of the SON was performed during late luteal phase. Our results suggest that in the guinea pig the SON modulates ovulation, and that the degree of modulation varies along the estrous cycle. The strongest influence of the SONs on ovulation occurs during early luteal phase, and decrease thereafter, being absent by late luteal phase. In addition, sectioning the left or the right SON caused different responses by the ovaries of adult guinea pigs. This paper discusses the mechanisms by which ovulation increased when the SON was surgically cut. PMID- 14561220 TI - ADAMTS4 (aggrecanase-1) cleaves human brain versican V2 at Glu405-Gln406 to generate glial hyaluronate binding protein. AB - Human brain tissue from cerebellum and hippocampus was obtained between 2 h and 24 h post mortem and, after extraction in the presence of proteinase inhibitors, proteoglycans were purified by anion-exchange chromatography. The versican component was characterized by Western analysis with antibodies to the N-terminal peptide (LF99), the N-terminal globular domain (12C5) and the two GAG (glycosaminoglycan) attachment regions (anti-GAG-alpha and anti-GAG-beta). The results indicated that versican V2 is the major variant in all brain samples, and that it exists as the full-length form and also as at least six C-terminally truncated forms. The major immunoreactive species present is a 64 kDa product, which we identified by biochemical and immunological analysis as the brain protein previously termed GHAP (glial hyaluronate binding protein) [Perides, Lane, Andrews, Dahl and Bignami (1989) J. Biol. Chem. 264, 5981-5987]. Immunological analysis of purified human GHAP using a new anti-neoepitope antiserum (JSCNIV) showed that its C-terminal sequence is NIVSFE(405), and digestion of human cerebellum proteoglycans with ADAMTS4 (aggrecanase-1, where ADAMTS, a disintegrin and metalloproteinase with thrombospondin-1-like motifs) indicated that GHAP is a product of cleavage of versican V0 or V2 at the Glu(405) Gln(406) bond. Since human cerebellum extracts contained multiple forms of ADAMTS4 protein on Western analysis, these data suggest that one or more members of the 'aggrecanase' group of the ADAMTS family (ADAMTS 1, 4, 5 and 9) are responsible for turnover of versican V2 in the adult human brain. PMID- 14561224 TI - Inapparent twin malignancy in thyroglossal cyst: Case Report. AB - BACKGROUND: Although malignancy has been reported in thyroglossal cysts, synchronous occurrence of two malignancies is extremely rare. CASE REPORT: A case of concurrent papillary and squamous carcinoma arising in the thyroglossal cyst is presented here. CONCLUSIONS: Papillary and squamous carcinoma simultaneously occurring in a thyroglossal cyst is rarely diagnosed prior to surgery and pose a therapeutic dilemma. In view of the extreme rarity of the condition, controversies do exist regarding the optimal strategy to be adopted. The ideal procedure needs to be individually tailored and involve a combination of surgery, radio ablation, thyroid suppression and external radiotherapy. PMID- 14561226 TI - Geographic disparity in premature mortality in Ontario, 1992-1996. AB - BACKGROUND: Standardized mortality ratios are used to identify geographic areas with higher or lower mortality than expected. This article examines geographic disparity in premature mortality in Ontario, Canada, at three geographic levels of population and considers factors that may underlie variations in premature mortality across geographic areas. All-cause, sex and disease chapter specific premature mortality were analyzed at the regional, district and public health unit level to determine the extent of geographic variation. Standardized mortality ratios for persons aged 0-74 years were calculated to identify geographic areas with significantly higher or lower premature mortality than expected, using Ontario death rates as the basis for the calculation of expected deaths in the local population. Data are also presented from the household component of the 1996/97 National Population Health Survey and from the 1996 Statistics Canada Census. RESULTS: Results showed approximately 20% higher than expected all-cause premature mortality for males and females in the North region. However, disparity in all-cause premature mortality in Ontario was most pronounced at the public health unit level, ranging from 20% lower than expected to 30% higher than expected. Premature mortality disparities were largely influenced by neoplasms, circulatory diseases, injuries and poisoning, respiratory diseases and digestive diseases, which accounted for more than 80% of all premature deaths. Premature mortality disparities were also more pronounced for disease chapter specific mortality. CONCLUSION: Geographic disparities in premature mortality are clearly greater at the small area level. Geographic disparities in premature mortality undoubtedly reflect the underlying distribution of population health determinants such as health related behaviours, social, economic and environmental influences. PMID- 14561227 TI - Effect of non-linearity in predicting Doppler waveforms through a novel model. AB - BACKGROUND: In pregnancy, the uteroplacental vascular system develops de novo locally in utero and a systemic haemodynamic & bio-rheological alteration accompany it. Any abnormality in the non-linear vascular system is believed to trigger the onset of serious morbid conditions like pre-eclampsia and/or intrauterine growth restriction (IUGR). Exact Aetiopathogenesis is unknown. Advancement in the field of non-invasive doppler image analysis and simulation incorporating non-linearities may unfold the complexities associated with the inaccessible uteroplacental vessels. Earlier modeling approaches approximate it as a linear system. METHOD: We proposed a novel electrical model for the uteroplacental system that uses MOSFETs as non-linear elements in place of traditional linear transmission line (TL) model. The model to simulate doppler FVW's was designed by including the inputs from our non-linear mathematical model. While using the MOSFETs as voltage-controlled switches, a fair degree of controlled-non-linearity has been introduced in the model. Comparative analysis was done between the simulated data and the actual doppler FVW's waveforms. RESULTS & DISCUSSION: Normal pregnancy has been successfully modeled and the doppler output waveforms are simulated for different gestation time using the model. It is observed that the dicrotic notch disappears and the S/D ratio decreases as the pregnancy matures. Both these results are established clinical facts. Effects of blood density, viscosity and the arterial wall elasticity on the blood flow velocity profile were also studied. Spectral analysis on the output of the model (blood flow velocity) indicated that the Total Harmonic Distortion (THD) falls during the mid-gestation. CONCLUSION: Total harmonic distortion (THD) is found to be informative in determining the Feto-maternal health. Effects of the blood density, the viscosity and the elasticity changes on the blood FVW are simulated. Future works are expected to concentrate mainly on improving the load with respect to varying non-linear parameters in the model. Heart rate variability, which accounts for the vascular tone, should also be included. We also expect the model to initiate extensive clinical or experimental studies in the near future. PMID- 14561225 TI - The development and preliminary validation of a Preference-Based Stroke Index (PBSI). AB - BACKGROUND: Health-related quality of life (HRQL) is a key issue in disabling conditions like stroke. Unfortunately, HRQL is often difficult to quantify in a comprehensive measure that can be used in cost analyses. Preference-based HRQL measures meet this challenge. To date, there are no existing preference-based HRQL measure for stroke that could be used as an outcome in clinical and economic studies of stroke. The aim of this study was to develop the first stroke-specific health index, the Preference-based Stroke Index (PBSI). METHODS: The PBSI includes 10 items; walking, climbing stairs, physical activities/sports, recreational activities, work, driving, speech, memory, coping and self-esteem. Each item has a 3-point response scale. Items known to be impacted by a stroke were selected. Scaling properties and preference-weights obtained from individuals with stroke and their caregivers were used to develop a cumulative score. RESULTS: Compared to the EQ-5D, the PBSI showed no ceiling effect in a high-functioning stroke population. Moderately high correlations were found between the physical function (r = 0.78), vitality (r = 0.67), social functioning (r = 0.64) scales of the SF-36 and the PBSI. The lowest correlation was with the role emotional scale of the SF-36 (r = 0.32). Our results indicated that the PBSI can differentiate patients by severity of stroke (p < 0.05) and level of functional independence (p < 0.0001). CONCLUSIONS: Content validity and preliminary evidence of construct validity has been demonstrated. Further work is needed to develop a multiattribute utility function to gather information on psychometric properties of the PBSI. PMID- 14561229 TI - Possible mechanisms of hypotension produced 70% alcoholic extract of Terminalia arjuna (L.) in anaesthetized dogs. AB - BACKGROUND: The bark of Terminalia arjuna L. (Combretaceae) is used in Ayurveda since ancient times for the treatment of cardiac disorders. Previous laboratory investigations have demonstrated the use of the bark in cardiovascular complications. The present study was aimed to find the effect of 70% alcoholic extract of Terminalia arjuna on anaesthetized dog blood pressure and probable site of action. METHODS: Six dogs were anaesthetized with intraperitoneal injection of thiopental sodium and the blood pressure of each dog (n = 6) was measured from the left common carotid artery connected to a mercury manometer on kymograph. The femoral vein was cannulated for administration of drug solutions. The extract of T. arjuna (dissolved in propylene glycol) in the dose range of 5 to 15 mg/kg were administered intravenously in a pilot study and the dose (6 mg/kg) which produced appreciable hypotension was selected for further studies. RESULTS: Intravenous administration of T. arjuna produced dose-dependent hypotension in anaesthetized dogs. The hypotension produced by 6 mg/kg dose of the extract was blocked by propranolol but not by atropine or mepyramine maleate. This indicates that muscarinic or histaminergic mechanisms are not likely to be involved in the hypotension produced by the extract. The blockade by propranolol of the hypotension produced by T. arjuna indicates that the extract might contain active compound(s) possessing adrenergic beta2-receptor agonist action and/or that act directly on the heart muscle. CONCLUSION: The results indicated the likely involvement of peripheral mechanism for hypotension produced by the 70% alcoholic extract of Terminalia arjuna and lends support for the claims of its traditional usage in cardiovascular disorders. PMID- 14561228 TI - Surface pretreatments for medical application of adhesion. AB - Medical implants and prostheses (artificial hips, tendono- and ligament plasties) usually are multi-component systems that may be machined from one of three material classes: metals, plastics and ceramics. Typically, the body-sided bonding element is bone. The purpose of this contribution is to describe developments carried out to optimize the techniques, connecting prosthesis to bone, to be joined by an adhesive bone cement at their interface. Although bonding of organic polymers to inorganic or organic surfaces and to bone has a long history, there remains a serious obstacle in realizing long-term high bonding strengths in the in vivo body environment of ever present high humidity. Therefore, different pretreatments, individually adapted to the actual combination of materials, are needed to assure long term adhesive strength and stability against hydrolysis. This pretreatment for metal alloys may be silica layering; for PE-plastics, a specific plasma activation; and for bone, amphiphilic layering systems such that the hydrophilic properties of bone become better adapted to the hydrophobic properties of the bone cement. Amphiphilic layering systems are related to those developed in dentistry for dentine bonding. Specific pretreatment can significantly increase bond strengths, particularly after long term immersion in water under conditions similar to those in the human body. The bond strength between bone and plastic for example can be increased by a factor approaching 50 (pealing work increasing from 30 N/m to 1500 N/m). This review article summarizes the multi-disciplined subject of adhesion and adhesives, considering the technology involved in the formation and mechanical performance of adhesives joints inside the human body. PMID- 14561232 TI - High altitude medicine: a British perspective. PMID- 14561230 TI - Single amino acids in the carboxyl terminal domain of aquaporin-1 contribute to cGMP-dependent ion channel activation. AB - BACKGROUND: Aquaporin-1 (AQP1) functions as an osmotic water channel and a gated cation channel. Activation of the AQP1 ion conductance by intracellular cGMP was hypothesized to involve the carboxyl (C-) terminus, based on amino acid sequence alignments with cyclic-nucleotide-gated channels and cGMP-selective phosphodiesterases. RESULTS: Voltage clamp analyses of human AQP1 channels expressed in Xenopus oocytes demonstrated that the nitric oxide donor, sodium nitroprusside (SNP; 3-14 mM) activated the ionic conductance response in a dose dependent manner. Block of soluble guanylate cyclase prevented the response. Enzyme immunoassays confirmed a linear dose-dependent relationship between SNP and the resulting intracellular cGMP levels (up to 1700 fmol cGMP /oocyte at 14 mM SNP). Results here are the first to show that the efficacy of ion channel activation is decreased by mutations of AQP1 at conserved residues in the C terminal domain (aspartate D237 and lysine K243). CONCLUSIONS: These data support the idea that the limited amino acid sequence similarities found between three diverse classes of cGMP-binding proteins are significant to the function of AQP1 as a cGMP-gated ion channel, and provide direct evidence for the involvement of the AQP1 C-terminal domain in cGMP-mediated ion channel activation. PMID- 14561234 TI - Electron paramagnetic resonance spectroscopic evidence of increased free radical generation and selective damage to skeletal muscle following lightning injury. AB - The present case study examined changes in peripheral markers of free radical metabolism and skeletal/myocardial muscle damage 30 h after a mountaineer had survived a lightning storm, having experienced contact with what was considered to be "upward leaders" at 4200 m. Sea-level control data were available between 3 and 8 weeks prior to the altitude sojourn for comparative purposes. Follow-up measurements were obtained for the same individual 3 weeks following the incident after simulated exposure to the combined stresses of inspiratory hypoxia and physical exercise. Venous blood was assayed for molecular markers of skeletal [myoglobin and total creatine phosphokinase (CPK)] and myocardial [cardiac troponin I (cTnI)] muscle damage. Ex-vivo spin trapping with alpha-phenyl-tert butylnitrone (PBN) combined with electron paramagnetic resonance (EPR) spectroscopy was incorporated for the direct detection of free radicals. The relative increases [post-exposure/preexposure x 100 (%)] in the concentration of the PBN adduct, myoglobin, and CPK in the "lightning blood" were markedly greater than those observed following the simulation study (PBN: 276 vs. 129%; CPK: 1130 vs. 182%; myoglobin: 205 vs. 115%). In contrast, no changes were observed for cTnI. A marked decrease in the PBN adduct, myoglobin, and CPK was observed within 2 h of completing the simulation study, following oral administration of water and lipid-soluble antioxidant vitamins in normoxia. These findings are the first to document lightning-induced free radical generation and selective damage to skeletal muscle in a high altitude mountaineer. Furthermore, free radicals may contribute to the pathogenesis of lightning injury, and dietary supplementation with antioxidant vitamins may prove of some benefit against associated tissue damage. PMID- 14561231 TI - Clinical and histological features of nonalcoholic steatohepatitis in Iranian patients. AB - BACKGROUND: Although several studies have been performed on risk factors and natural course of NASH, it seems that NASH tends to be more than a disease confined to strict boundaries. The objective of this study was to assess the clinical and paraclinical features and risk factors for non-alcoholic steatohepatitis (NASH) patients in an Iranian population METHODS: Patients with histologically confirmed NASH who had elevated liver aminotransaminases, negative serologic markers of viral or autoimmune hepatitis and no findings in favor of metabolic liver disease were enrolled. A careful history was taken regarding alcohol intake. RESULTS: 53 patients consisting of 32 male and 21 female entered the study. The mean age was 37.8 +/- 11.3 years. Twenty-six patients (55.3%) were overweight, 15 (31.9%) obese, 40 (75.5%) dyslipidemic, and three patients (5.7%) were diabetic. Liver biopsy showed mild steatosis in 35.7%, moderate steatosis in 53.6%, and severe forms in 10.7%. In 80.2% of patients, portal inflammation was present, and 9.4% had cirrhosis. The amount of increase in liver enzymes bore no relationship with fibrosis, portal inflammation, and degree of steatosis. CONCLUSIONS: The patients in our study showed a male predominancy and were somewhat younger than other studies. PMID- 14561235 TI - Intermittent hypoxia improves endurance performance and submaximal exercise efficiency. AB - The purpose of the present study was to elucidate the influence of intermittent hypobaric hypoxia at rest on endurance performance and cardiorespiratory and hematological adaptations in trained endurance athletes. Twelve trained male endurance runners were assigned to either a hypoxic group (n = 6) or a control group (n = 6). The subjects in the hypoxic group were exposed to a simulated altitude of 4500 m for 90 min, three times a week for 3 weeks. The measurements of 3000 m running time, running time to exhaustion, and cardiorespiratory parameters during maximal exercise test and resting hematological status were performed before (Pre) and after 3 weeks of intermittent hypoxic exposure (Post). These measurements were repeated after the cessation of intermittent hypoxia for 3 weeks (Re). In the control group, the same parameters were determined at Pre, Post, and Re for the subjects not exposed to intermittent hypoxia. The athletes in both groups continued their normal training together at sea level throughout the experiment. In the hypoxic group, the 3000 m running time and running time to exhaustion during maximal exercise test improved. Neither cardiorespiratory parameters to maximal exercise nor resting hematological parameters were changed in either group at Post, whereas oxygen uptake (.V(O2)) during submaximal exercise decreased significantly in the hypoxic group. After cessation of intermittent hypoxia for 3 weeks, the improved 3000 m running time and running time to exhaustion tended to decline, and the decreased .V(O2) during submaximal exercise returned to Pre level. These results suggest that intermittent hypoxia at rest could improve endurance performance and submaximal exercise efficiency at sea level in trained endurance athletes, but these improvements are not maintained after the cessation of intermittent hypoxia for 3 weeks. PMID- 14561236 TI - Effects of hypoxia on the circadian patterns in men. AB - We tested the hypothesis that acute hypoxia may alter the circadian pattern of body temperature in adult humans. Six healthy subjects were studied in normoxia, hypoxia (approximately 13% inspired O(2)), and again normoxia, each session lasting >24 h and spaced a few days apart, with a constant routine protocol of sustained wakefulness and minimal activity. Some parameters (e.g., tympanic and abdominal temperatures, heart rate) were recorded continuously; others (e.g., oxygen consumption and pulmonary ventilation) were monitored for approximately 10 min every 2 h. The amplitudes of the circadian oscillation of tympanic, abdominal, and calf skin temperatures were reduced in hypoxia, averaging, respectively, 61%, 80% and 50% of the normoxic amplitude. Oxygen consumption and pulmonary ventilation, which presented a circadian pattern in normoxia, had no longer significant oscillations during hypoxia, whereas the opposite was the case for heart rate and diastolic pressure. Therefore, acute hypoxia can disturb the normal circadian patterns and, specifically, depress those of body temperature. These effects, qualitatively similar to those observed in chronically hypoxic animals and humans, could contribute to sleep disturbances at high altitude. PMID- 14561237 TI - Symptoms of infection and acute mountain sickness; associated metabolic sequelae and problems in differential diagnosis. AB - Infections and acute mountain sickness (AMS) are common at high altitude, yet their precise etiologies remain elusive and the potential for differential diagnosis is considerable. The present study was therefore designed to compare clinical nonspecific symptoms associated with these pathologies and basic changes in free radical and amino-acid metabolism. Nineteen males were examined at rest and after maximal exercise at sea level before (SL(1)/SL(2)) and following a 20 +/- 5 day ascent to Kanchenjunga base camp located at 5100 m (HA). Four subjects with symptoms consistent with an ongoing respiratory and recent gastrointestinal infection were also diagnosed with clinical AMS on the evening of day 1 at HA. These and six other subjects recovering from symptoms consistent with a respiratory infection presented with a greater increase (HA minus SL(1)) in AMS scores and resting venous concentration of lipid hydroperoxides (LH) and in total creatine phosphokinase and ratio of free tryptophan/branched chain amino acids, and greater decrease in glutamine (Gln) compared to healthy controls (n = 9, p < 0.05). The decrease in Gln was consistently related to the altitude/exercise induced increase in LH (r = -0.69/r = -0.45; p < 0.05) and altitude-induced increase in myoglobin (r = -0.73, p < 0.05). These findings highlight the potential for the misdiagnosis of altitude illness due to the similarity of nonspecific constitutional symptoms associated with infection and AMS. Both conditions were characterized by parallel changes in peripheral biomarkers related to free-radical, skeletal muscle damage and amino acid metabolism. While clearly not establishing cause and effect, free radical-mediated changes in peripheral amino acid metabolism known to influence immune and cerebral serotoninergic function may enhance susceptibility to and/or delay recovery from altitude illness. PMID- 14561238 TI - Flunarizine in prevention of headache, ataxia, and memory deficits during decompression to 4559 m. AB - Our purpose was to study the preventive effect of the calcium channel blocker flunarizine on headache, postural ataxia, and memory deficits occurring during decompression to high altitude in a randomized, placebo-controlled, double-blind study. After 7-day pretreatment with the study drugs, 20 healthy men were investigated at 490 m and 0.5, 2, 4, and 6 h later at a simulated altitude of 4559 m. Headache severity was evaluated on a 4-point scale. Sway path and anteroposterior and lateral sway were recorded with open and closed eyes by static posturography. Short- and long-term memory was studied by testing the recall of verbal and figural material immediately and 2 h after presentation, respectively. Blood pressure (BP) and arterial oxygen saturation (Sa(O2)) were also assessed. Headache scores showed a trend to be lower in the flunarizine group that was significant after 4 and 6 h. Headache scores expressed as difference from baseline values showed a nonsignificant trend to be lower at 4 and 6 h in subjects treated with flunarizine. Postural stance, memory, BP, and Sa(O2) were similar in both treatment groups. Although the low number of investigated subjects may have prevented the detection of a significant therapeutic effect of flunarizine, the present data do not show that flunarizine is effective for prevention of headache, postural ataxia, and neurocognitive deficits occurring at simulated high altitude. PMID- 14561239 TI - Hypoxic ventilatory response changes of men and women 6 to 7 days after climbing from 2100 m to 4350 m altitude and after descent. AB - To test the hypothesis that the changes in hypoxic ventilatory response (HVR) of men and women mountaineers on induction to HA by trekking is not influenced by gender, isocapnic HVR as DeltaV(E)/DeltaSa(O2) was studied in eight men and eight women mountaineering trainees initially at 2100 m, then during 6 to 7 days of sojourn at 4350 m, and retested again on return to 2100 m. Results indicated that HVR at 2100 m increased significantly at 4350 m in both sexes, and the values reverted to baseline level within 4 to 5 days between leaving high altitude (4350 m) and restudy at 2100 m. No sex differences were observed at 2100- or at 4350-m altitude, indicating that men and women have a similar level of chemosensitive response as measured by HVR during induction to HA. PMID- 14561240 TI - Changes in male reproductive function after high altitude mountaineering. AB - The male reproductive functions of the members of the Masherbrum (7821 m) Expedition in 1999 were examined via semen analyses and endocrine tests. Specimens were collected from three subjects who had stayed above 5100 m for 21 to 24 days and above 6700 m for 4 to 5 days before departure and 1 month, 3 months, and 2 yr after returning from the expedition. Semen analyses showed no change in the semen volume. Sperm counts decreased after 1 month and had not recovered after 3 months, but they had recovered after 2 yr in all subjects. An increase in abnormally shaped sperm was also observed after 1 month, but had nearly recovered to the preexpedition state after 3 months. Endocrine tests revealed slightly decreased testosterone in the blood after 1 month, which had decreased still further after 3 months. The tests were completely normal after 2 yr. We suggest that a high altitude sojourn may induce reversible spermatogenic and Leydig cell dysfunction. PMID- 14561241 TI - Lack of evidence for association of high altitude pulmonary edema and polymorphisms of the NO pathway. AB - One essential factor in the development of high altitude pulmonary edema (HAPE) is elevated pulmonary artery pressure, possibly due to a lack of nitric oxide (NO) in pulmonary vessels. NOS3 gene polymorphisms (G894T, T-786C, and CA-repeats > or =38) might be linked to decreased NO synthesis and increased susceptibility to HAPE, while the C242T polymorphism of the CYBA gene [encoding for the NAD(P)H oxidase subunit p22phox] may increase NO availability and thus convey resistance to HAPE. To test this hypothesis, we genotyped 51 mountaineers susceptible and 52 mountaineers not susceptible to HAPE. Genotyping revealed similar genotype frequencies of the G894T and the T-786C NOS3 polymorphism in both groups (G894T: susceptibles, 39.2% GG, 47.1% GT, 13.7% TT; nonsusceptibles, 48.0% GG, 44.0% GT, 8.0% TT; p = 0.54. T-786C: susceptibles, 45.1% TT, 39.2% TC, 15.7% CC; nonsusceptibles, 53.8% TT, 40.4% TC, 5.8% CC; p = 0.28). Genotype frequencies of the C242T CYBA polymorphism were 43.1% CC, 47.1 % CT, and 9.8% TT in HAPE susceptibles and 38.0% CC, 52.0 % CT, and 10.0% TT (p = 0.92) in nonsusceptibles. There was also no difference between the two groups in the number of CA repeats (p = 0.57), and individuals with > or =38 CA repeats were not more likely to develop HAPE (p = 1.0). Haplotype analysis for the NOS3 polymorphisms also revealed no association with HAPE. The results of this study suggest that none of these genetic variants plays a substantial role in the pathogenesis of HAPE in Caucasians, but does not exclude epistatic effects that might still involve the genetic systems studied here. PMID- 14561243 TI - Heart rate variability in rats acclimatized to high altitude. AB - The aims of this study were to relate heart morphology and functions changes to heart rate variability (HRV) components after acclimatization to high altitude and to define whether preadaptation to hypoxia could modulate HRV responses to acute hypoxic stress. Doppler-echocardiographic studies of the left ventricle were performed in female Wistar rats before, during, and after a 10-week exposure to moderate hypobaric hypoxia (CH rats, approximately 4000 m simulated) or normoxia (N rats, approximately 55 m). Right ventricular morphology and function and pulmonary artery pressure were evaluated using heart catheterization. Spectral analysis of HRV was studied after exposure in conscious unrestrained rats in normoxia and during acute hypoxic stress. Necropsy right ventricular hypertrophy and intraventricular and pulmonary artery hypertension were found in CH rats compared with N rats. Echocardiographic left ventricular morphology and functions were similar between the groups after exposures. Compared to the control group, CH rats had similar heart rates and HRV components when measured in normoxia. During acute hypoxic stress, HRV decreased in all rats, but less in CH rats. These results support the hypothesis that long-term mild hypoxia may moderate sympathetic activation induced by acute hypoxia and that right ventricular hypertrophy cannot be the direct cause of such a shift in sympathovagal nerve interaction during acute hypoxic stress. PMID- 14561242 TI - Body temperature, autonomic responses, and acute mountain sickness. AB - A few studies have reported increased body temperature (T(o)) associated with acute mountain sickness (AMS), but these usually include exercise, varying environmental conditions over days, and pulmonary edema. We wished to determine whether T(o) would increase with AMS during early exposure to simulated altitude at rest. Ninety-four exposures of 51 men and women to reduced P(B) (423 mmHg = 16,000 ft = 4850 m) were carried out for 8 to 12 h. AMS was evaluated by LL and AMS-C scores near end of exposure, and T(o) was measured by oral digital thermometer before altitude and after 1 (A1), 6 (A6), and last (A12) h at simulated altitude. Other measurements included ventilation, O(2) consumption and autonomic indicators of plasma catecholamines, HR, and HR variability. Average T(o) increased by 0.5 degrees F from A1 to A12 in all subjects (p < 0.001). Comparison between 16 subjects with lowest AMS scores (mean LL = 1.0, range = 0 to 2.5) and 16 other subjects with highest AMS scores (mean LL = 7.4, range = 5 to 11) demonstrated a transient decline in T(o) from A1 to A6 in AMS, in contrast to a rise in non-AMS (p = 0.001). Catecholamines, HR, and HR variability (increased low F/high F ratio) indicated significant elevation of sympathetic activity in AMS, where T(o) fell, but no change in metabolic rate. The apparently greater heat loss during early AMS suggests increased hypoxic vasodilation in spite of enhanced sympathetic drive. Greater hypoxic vasodilation and elevated HR in AMS in the absence of other changes suggest that augmentation of beta adrenergic tone may be involved in early AMS pathophysiology. PMID- 14561244 TI - Improving oxygenation at high altitude: acclimatization and O2 enrichment. AB - When lowlanders go to high altitude, the resulting oxygen deprivation impairs mental and physical performance, quality of sleep, and general well-being. This paper compares the effects of ventilatory acclimatization and oxygen enrichment of room air on the improvement of oxygenation as judged by the increase in the alveolar P(O2) and the reduction in equivalent altitude. The results show that, on the average, complete ventilatory acclimatization at an altitude of 5000 m increases the alveolar P(O2) by nearly 8 torr, which corresponds to a reduction in equivalent altitude of about 1000 m, although there is considerable individual variability. By comparison, oxygen enrichment to 27% at 5000 m can easily reduce the equivalent altitude to 3200 m, which is generally well tolerated. Because full ventilatory acclimatization at altitudes up to about 3600 m reduces the equivalent altitude to about 3000 m, oxygen enrichment is not justified for well acclimatized persons. At an altitude of 4200 m, where several telescopes are located on the summit of Mauna Kea, full acclimatization reduces the equivalent altitude to about 3400 m, but the pattern of commuting probably would not allow this. Therefore, at this altitude, oxygen enrichment would be beneficial but is not essential. At higher altitudes such as 5050 m, where other telescopes are located or planned, the gain in oxygenation from acclimatization is insufficient to produce an adequate mental or physical performance for most work, and oxygen enrichment is highly desirable. Full ventilatory acclimatization requires at least a week of continuous exposure, although much of the improvement is seen in the first 2 days. PMID- 14561245 TI - Efficacy of low-dose acetazolamide for the prophylaxis of acute mountain sickness. PMID- 14561248 TI - The surgical treatment of obesity: a personal perspective. PMID- 14561249 TI - Mechanisms of weight loss after bariatric surgery. AB - The surgical treatment of clinically severe obesity is becoming more popular. To date, little has been published concerning the mechanisms by which each of the commonly employed bariatric surgical procedures induces weight loss. This article reviews the physiology of weight loss induced by semi-starvation and other proposed mechanisms of surgically induced weight loss. PMID- 14561250 TI - Evolution of the laparoscopic gastric bypass. AB - Obesity is recognized as a health problem of epidemic proportions. Surgical intervention for the treatment of obesity is a well-studied and effective method. Various procedures have been utilized over the past decades. Roux-en-Y gastric bypass has emerged over the last 20 years and is currently the most commonly offered surgical treatment. Within the last decade, advances in laparoscopic technology and surgical experience have allowed the application of laparoscopic techniques to the surgical treatment of obesity. Many centers and individuals have developed excellent techniques through experience over time as well as improvements in instrumentation. Hand-assisted laparoscopy was reported as a technique, but has mostly fallen out of favor. Currently, laparoscopic application of adjustable gastric band and laparoscopic Roux-en-Y gastric bypass are widely used throughout the United States. Data have been generated to demonstrate the improvement in surgical outcomes associated with minimally invasive surgical techniques for the surgical treatment of obesity. Further advances will allow continued improvement in patient outcomes utilizing a variety of minimally invasive surgical approaches to the treatment of this difficult disease. PMID- 14561251 TI - Laparoscopic gastric bypass surgery: current technique. PMID- 14561252 TI - Laparoscopic gastric bypass surgery: equipment and necessary tools. AB - Advanced laparoscopy in the morbidly obese patient is technically challenging. Having the proper instrumentation and equipment available is a major component of technical success. Items routinely used during surgery performed on patients of normal size must often be modified or substituted when morbidly obese patients undergo surgery. In this article, we review the specific tools necessary for the safe and proper completion of laparoscopic Roux-en-Y gastric bypass, in addition to various alternatives that can be helpful when other procedures are performed on morbidly obese patients. PMID- 14561253 TI - Laparoscopic gastric bypass surgery: outcomes. PMID- 14561254 TI - Laparoscopic adjustable gastric banding: surgical technique. AB - Laparoscopic adjustable gastric banding is an effective and safe surgical treatment for morbid obesity. Initial experience with the Lap-Band system (Inamed Health, Santa Barbara, California) in the United States and Australia has demonstrated that surgical technique can affect outcomes in terms of weight loss, quality of life, and complication rates. Placement of the gastric band by means of the perigastric technique is associated with high rates of gastric prolapse, food intolerance, and weight loss failure that frequently lead to band explantation. In the pars flaccida technique, band placement higher on the stomach results in the formation of a smaller pouch and lower rates of gastric prolapse, which may contribute to greater weight loss and improved quality of life. This article describes the technical aspects of the pars flaccida approach in the laparoscopic placement of adjustable gastric bands. PMID- 14561255 TI - Lap-band: outcomes and results. AB - INTRODUCTION: Laparoscopic adjustable gastric banding was first introduced in the early 1990s as a potentially safe, controllable, and reversible method for achieving significant weight loss in the severely obese. The Bioenterics Lap-Band system (Inamed Health, Santa Barbara, California) is the device most commonly used. After 10 years of experience in treating more than 100000 patients with the Lap-Band, it is timely for us to review the outcomes. METHODS: Data for the review are derived from the experience of our unit in the treatment of 1250 patients to date, from an independent systematic review of the published literature up to September 2001, and from major studies published after the date of closure of the systematic review. RESULTS: Lap-Band placement has proved to be a very safe procedure with a mortality rate in the published reports of 1 in 2000, only 10% of the published mortality rate of gastric bypass. The early complication rate has been very low, but late complications of prolapse or erosions have been more frequent, particularly during the early experience. Weight is lost during the first 2 to 3 years after surgery. The systematic review reports 56% excess weight loss (EWL) at 5 years (three reports). In comparison, Roux-en-Y gastric bypass (RYGB) is reported to have achieved 59% EWL at 5 years (four reports). Major improvements in comorbid conditions have been reported in association with weight loss after Lap-Band placement. Most importantly, type 2 diabetes is usually cured, and insulin resistance and reduced pancreatic beta cell function are reversed. Gastroesophageal reflux, obstructive sleep apnea, and depression are other diseases in which marked improvement is noted. Quality-of life scores return to normal values. CONCLUSIONS: Lap-Band placement is proving to be safe and effective. In view of the attributes of adjustability, safe laparoscopic placement, and reversibility, it should be considered the optimal initial approach for the control of obesity and its comorbid conditions. PMID- 14561256 TI - Laparoscopic adjustable silicone gastric banding: complications. AB - Laparoscopic adjustable gastric banding is a procedure that is now approved by the Federal Drug Administration for use in the United States to treat morbid obesity. Numerous complications can occur as a result of the device. These include both early technical complications as well as long-term problems such as esophageal dilatation and failed weight loss. While improvements in surgical technique may decrease early technical complications such as gastric prolapse, long-term follow-up studies will be required to determine the ultimate success of this device in controlling severe obesity. PMID- 14561257 TI - The role of robotic surgery in morbid obesity. AB - The purpose of this review was to form a collaborative opinion regarding the utility of robotic technology, specifically the da Vinci robot (Intuitive Surgical, Inc., Sunnyvale, CA), for bariatric surgery. Eleven surgeons are currently using the da Vinci system to perform laparoscopic bariatric surgery. The surgeons were queried about the type of robotic bariatric surgery and number of cases performed, complications, and any available outcomes data. They were also asked to give their opinion of the current state of robotic bariatric surgery. Ten of the 11 surgeons responded. Six surgeons are currently using the system for Roux-en-Y gastric bypass, three for laparoscopic adjustable gastric banding, and three for biliary pancreatic diversion. Robotic technology is successfully being applied to bariatric surgery in a few advanced minimally invasive surgery centers throughout the United States. PMID- 14561258 TI - Obesity surgery: a gastroenterologist's perspective. AB - Obesity is increasing at an alarming rate. Approximately 25% of adult women and 20% of adult men in the United States are obese. Obesity is increasing even more rapidly in children. The incidence of type 2 diabetes mellitus, hypertension, dyslipidemia, and liver disease is significantly increased in obese persons. Traditional methods of diet, exercise, drugs, and behavior modification are unsuccessful in the treatment of patients who are morbidly obese and have a body mass index of 40 kg/m(2) or a body mass index of 35 kg/m(2) with comorbidity. Multiple surgical alternatives to the traditional treatments are available and have been successful. Considerable weight loss may be achieved and maintained. Each procedure is associated with a variety of side effects and complications. The selection of patients for bariatric surgery requires a careful evaluation of their medical condition in addition to multiple psychological and social factors. Postoperative care entails careful medical follow-up and long-term support. Patients may have a difficult time adjusting to their new ability to eat normally. PMID- 14561259 TI - Prenatal screening for cystic fibrosis in the United States -- time to re evaluate implementation policies. PMID- 14561260 TI - Maternal serum screening in Ontario using the triple marker test. AB - OBJECTIVES: To summarise the experience and evaluate the performance of the Ontario maternal serum screening (MSS) programme. SETTING: The Ontario MSS programme between October 1993 and September 2000. METHODS: This study used information collected in the Ontario MSS database, which contains data on each screened pregnancy. In the Ontario MSS programme, women are screened between 15 and 20 weeks of gestation. The risk cut-off for Down's syndrome was >or= 1 in 385 at term and women with a serum alpha-fetoprotein >or= 2.2 multiples of the unaffected population median were defined as screen-positive for open neural tube defects. RESULTS: Between 1 October 1993 and 30 September 2000, 428410 women residing in Ontario were screened for open neural tube defects, and 423895 women were screened for Down's syndrome and trisomy 18. Approximately 48% of all pregnant women in the province had MSS. The uptake rate of amniocentesis following a positive Down's syndrome screening was 67%. Of 717 cases of Down's syndrome ascertained in the screened population, 531 were detected by MSS, giving a term detection rate (DR) of 70.6%, with a false-positive rate (FPR) of 7.2%. For neural tube defects, the DR was 72.7%, with a FPR of 2.0%. The screen also detected 50% of cases of trisomy 18 at term, with a FPR of 0.2%. Coincidentally, 113 cases of chromosome aneuploidies other than Down's syndrome and trisomy 18 were detected. DISCUSSION: In the Ontario MSS programme, MSS performed as expected in the detection of Down's syndrome, open neural tube defects and trisomy 18. MSS is an effective and practical method for large-scale second trimester screening for Down's syndrome, open neural tube defects and trisomy 18, and the MSS database is an extremely useful tool in monitoring the performance of this screen. PMID- 14561261 TI - Population screening for neonatal liver disease: potential for a community-based programme. AB - OBJECTIVES: To evaluate a community-based screening programme for detecting neonatal liver disease by the quantitation of conjugated bilirubin in blood. SETTINGS AND METHODS: Prospective cohort/observational study using spare plasma from routinely collected liquid neonatal screening specimens from babies born in Birmingham over a two-year period. Babies with a conjugated bilirubin above 18 mumol/l and comprising more than 20% of the total bilirubin were followed up. A total of 27654 neonates were tested in the community, with a further 2425 samples from babies hospitalised at the time of the test. RESULTS: In the community-based series, 84.7% of the specimens received were analysed, the remainder being unusable mainly because of gross haemolysis (8.6%) or insufficient sample (5.8%). In 107 neonates the results were above the cut-off limits (0.46% of the number analysed). Of these, 12 had persistently abnormal results, 11 of whom had confirmed liver disease. The liver diseases detected included neonatal hepatitis (n=6), extra-hepatic biliary atresia (n=2), hypopituitarism (n=1), alpha-1 antitrypsin deficiency (n=1) and Alagille syndrome (n=1). The sensitivity and specificity of the test for babies in the community were 100% and 99.6%, respectively. CONCLUSIONS: Conjugated bilirubin in plasma measured at 6-10 days is a reliable marker for neonatal liver disease, and a population screening programme based on this method has the potential to improve the survival and quality of life of infants born with liver disease. However, testing as part of the neonatal screening programme will prove practical only if the method can be adapted to use dried blood spots. PMID- 14561262 TI - A randomised trial of the impact of new faecal haemoglobin test technologies on population participation in screening for colorectal cancer. AB - OBJECTIVES: To investigate the effect on participation in colorectal cancer screening of testing for blood products in faeces using technologies that remove dietary restrictions (i.e. immunochemical tests) and simplify faecal sampling (i.e. tests that use brush sampling). METHODS: SETTING: Urban residents (n=1818) of Adelaide, Australia, aged between 50 and 69 years, randomly selected from the electoral roll. DESIGN: Three randomised cohorts of 606 invitees were offered a screening test by mail in 2001. The Hemoccult SENSA and FlexSure OBT cohorts were instructed to sample three stools using a spatula while the InSureTM cohort sampled two stools using a brush. The Hemoccult SENSA cohort was asked to restrict certain (high-peroxidase) foods and drugs. MAIN OUTCOME MEASURES: Participation (i.e. return of completed sample kits within 12 weeks) and generalised linear modelling (GLM) of relationships between participation, test technologies and demographic variables. RESULTS: Participation was 23.4%, 30.5% and 39.6% for the Hemoccult, FlexSure and InSure cohorts, respectively (chi(2)=37.1, p<0.00001). GLM demonstrated that participation was increased by 28% by removal of restrictions (p=0.01) and by 30% by simplification of sampling (p=0.001); both together increased participation by 66% (p<0.001). The differences in participation between tests occurred in the first three weeks. Socio-economic status, gender or age did not significantly influence technology based improvements in participation. CONCLUSIONS: The brush-sampling faecal immunochemical test for haemoglobin (InSure) achieves the best participation rates by simplifying sampling and removing the need for restrictions of diet and drugs. Because participation in screening is vital to detection, this new technology should contribute to better detection of neoplasia at the population level. PMID- 14561263 TI - Prescreening evaluation of a brush-based faecal immunochemical test for haemoglobin. AB - OBJECTIVES: To undertake a prescreening evaluation of a new brush-based faecal immunochemical test for haemoglobin, relative to a traditional spatula-sampling immunochemical test. METHODS: SETTING: Patients aged between 24 and 90 years, scheduled to undergo diagnostic colonoscopy in two major urban hospitals, for a range of clinical indications. DESIGN: Patients sampled three stools using a spatula for the reference FlexSure OBT test and two stools using a brush for the InSure test; order of sampling was randomised. Faecal haemoglobin was quantified by a modified InSure in a subset of patients to determine whether brush-sampling allowed discrimination between groups. MAIN OUTCOME MEASURES: Sensitivity for cancer or adenoma; false-positive rate in normals. Faecal haemoglobin levels. Preference for sampling method. RESULTS: InSure and FlexSure OBT did not differ in their sensitivities for cancer (27/36, 75% vs 29/36, 80.5%, respectively), adenomas >or= 10 mm (12/29, 41.4% vs 13/29, 44.8%) or adenomas <10 mm (each 8/56, 14.3%). Likewise, false-positive rates in normals were similar: 4/179 (2.2%) and 5/179 (2.8%) respectively (specificities of 97.8% and 97.2%, respectively). Levels of faecal haemoglobin were highest in those with cancers; those with adenomas had intermediate levels which were also significantly higher than those in normals. The brush sampling method was preferred by 38/46 (82.6%), while 4/46 (8.7%) preferred the spatula (p<0.00001). CONCLUSIONS: InSure is as sensitive and specific as FlexSure OBT for faecal haemoglobin. The novel stool-sampling method of InSure allows discrimination between normals and classes of neoplasia, and is highly preferred. The brush-sampling faecal immunochemical test InSure should now be evaluated in a screening population. PMID- 14561264 TI - False-positive result and reattendance in the Ontario Breast Screening Program. AB - OBJECTIVE: To determine the association between initial screen result and returning for a second screen in an organised breast screening programme for women with a biennial screening recommendation. SETTING: Women who attended the Ontario Breast Screening Program (OBSP). METHODS: A retrospective cohort study was conducted of 140723 Ontario women aged 50 years and older who had an initial screen at the OBSP between 1 July 1990 and 31 December 1995 and were followed until 30 June 1998. Rescreening rates at 36 months and risk ratio estimates were calculated using survival methods. Age of women, year of screen, region (within Ontario) and initial screen result were compared. For initial screen results, returning for a second screen was examined by integration of screening centre with an assessment programme and by modality of referral. RESULTS: Women with a false-positive result were less likely to return for a second screen as were women aged 70 and older and those living in regions of Ontario with fewer OBSP screening centres. However, there were minimal differences in reattendance behaviour by initial screen result for women screened at the OBSP centre with an assessment programme. CONCLUSIONS: Integration of breast screening and assessment services improved reattendance of women with false-positive screen results within an organised screening programme. PMID- 14561265 TI - Integrating self-referral for mammography into organised screening: results from an Italian experience. AB - OBJECTIVE: To compare self-referred screenees with respondents to invitation for main performance indicators of mammography screening. SETTING: First round of an organised, population-based screening programme in six districts of northern Italy. METHODS: The screening test was a two-view mammography. Eligible women aged 50-69 years were invited. Self-referred attendees were accepted if they were eligible for screening and had not yet been invited or had been invited >6 months before presentation. Age-specific performance indicators were compared with the calculation of their ratio and 95% confidence intervals (CIs). Total ratios were age standardised. In situ carcinomas were excluded. RESULTS: The eligible population was 183 542 women. There were 112 188 respondents to invitation and 20 280 self-referred attendees. Self-referral rate was inversely related to age. Performance indicators were as follows: recall rate, 5.6% for self-referred attendees vs 5.5% for respondents (ratio 1.02, 95% CI 0.96 to 1.08); total aspiration cytology rate, 37.3% vs 28.3% (1.37, 1.24 to 1.51); biopsy rate, 17.0 vs 12.6 x 1000 (1.51, 1.35 to 1.67); total detection rate, 10.7 vs 7.5 x 1000 (1.70, 1.48 to 1.94); detection rate of pT1 carcinoma, 7.0 vs 6.1 x 1000 (1.35, 1.14 to 1.59); detection rate of pT2-4 carcinoma, 3.5 vs 1.2 x 1000 (3.51, 2.75 to 4.43); false-positive rate, 4.5% vs 4.7% (0.93, 0.87 to 0.99); positive predictive value (PPV) of mammography, 19.1% vs 13.5% (1.59, 1.39 to 1.82); PPV of biopsy, 63.7% vs 60.6% (1.13, 0.98 to 1.29); detected:expected ratio, 5.02 vs 3.37 (1.49, 1.28 to 1.74). All differences were more pronounced among or restricted to women aged 50-54 years. CONCLUSIONS: Self-referred screenees were similar to respondents to invitation in main indicators of screening feasibility such as recall rate and PPV of biopsy, while showing important increases in detection rates and detected:expected ratios, especially among women aged 50-54 years. PMID- 14561266 TI - Lack of association between breast artery calcification seen on mammography and coronary artery disease on angiography. AB - OBJECTIVES: Some studies have found correlations between the presence of breast artery calcium (BAC) observed on routine mammograms and risk factors for coronary artery disease (CAD). The aim of this study was to investigate whether such calcifications could predict the presence of coronary atherosclerosis. METHODS: A total of 319 female patients between 50 and 70 years of age, 187 with significant CAD and 132 with angiographically normal coronary arteries, were randomly selected from a computerised database of our central catheterisation laboratory. The patients' mammograms were evaluated independently for the presence of BAC in a blinded fashion by an experienced breast radiologist, and additional clinical data were extracted from clinical charts. RESULTS: The women in the CAD group were older (62.5 vs 60.7 years, p=0.05) and had a higher prevalence of hypertension, diabetes mellitus and dyslipidaemia. Although the prevalence of BAC was marginally higher in the CAD group (43.9% vs 37.1%, p=0.138), this tendency was eliminated after controlling for confounders. Multiple regression analyses indicated that only age above 63 years (odds ratio [OR]=3.0, 95% confidence interval [CI]=1.8-4.9) and hypertension (OR=2.2, 95% CI=1.2-4.1), but not angiographic evidence of CAD (OR=1.0, 95% CI=0.6-1.6), predict with BAC on mammography. CONCLUSIONS: Despite correlation with some risk factors for CAD, the presence of BAC does not differentiate between patients with angiographic evidence of CAD and those with angiographically normal coronary arteries. PMID- 14561267 TI - Colposcopy information leaflets: what women want to know and when they want to receive this information. AB - OBJECTIVES: To evaluate whether the information leaflets produced by UK colposcopy clinics provide women with the information they desire and to determine when they would like to receive this information. DESIGN: Questionnaire study and structured evaluation. SETTING: The colposcopy clinic of a UK cancer centre. PARTICIPANTS: Forty-two women attending a pre-colposcopy counselling session and 100 consecutive women attending the colposcopy clinic. METHODS: Thirty-eight standards derived from the concerns/questions asked by women attending a pre-colposcopy counselling session were used to assess locally produced colposcopy clinic leaflets from UK colposcopy clinics, the leaflets produced by the Royal College of Obstetricians and Gynaecologists and the National Health Service Cervical Screening Programme (NHSCSP), and two "leaflets" obtained from internet sites. The Gunning fog test was used to assess the leaflets' readability. A questionnaire survey of 100 women attending the colposcopy clinic was used to determine when women wanted to receive information about colposcopy. MAIN OUTCOME MEASURES: Percentage of questions answered by a given leaflet and Gunning fog scores for readability. RESULTS: The information leaflets of 128 colposcopy clinics were received and assessed. Thirty-two clinics only sent women the NHSCSP leaflet. No leaflet answered all 38 questions. Less than half (36/100) of the leaflets answered more than 50% of the questions. In addition to the lack of advice given, different leaflets frequently gave conflicting advice. The average Gunning fog score was 9.7 (range 5.5-15.5). The majority of women (70%) wanted to receive information about colposcopy at or prior to the time of receiving their abnormal smear test result, although only 42% of women actually received information at this time. CONCLUSIONS: Many UK colposcopy clinics do not appear to be providing women with the information they require to understand their condition and the procedure that they are about to undergo. Furthermore, this information is often not provided at the appropriate time in the screening process. PMID- 14561268 TI - Cost-effectiveness of population screening for Helicobacter pylori in preventing gastric cancer and peptic ulcer disease, using simulation. AB - OBJECTIVES: To evaluate the cost-effectiveness of population screening for Helicobacter pylori in preventing gastric cancer and peptic ulcer disease in England and Wales. METHODS: A discrete event simulation model used parameter estimates, derived from peer-reviewed literature, routine data and statistical modelling. Population screening was compared with no screening but with opportunistic eradication in patients presenting with dyspepsia. Costs included screening, eradication and costs averted to provide costs per life years saved (cost/LYS) for preventing gastric cancer and peptic ulcer disease. Sensitivity analyses were undertaken. RESULTS: The cost/LYS from screening at age 40 years was Uk pounds 5860 at discount rates of 6%. The outcomes were sensitive to H. pylori prevalence, the degree of opportunistic eradication, the discount rate, the efficacy of eradication on gastric cancer risk, the risk of complicated peptic ulcer disease and gastric cancer associated with H. pylori infection, and the duration of follow-up. In sensitivity analyses, the cost/LYS rarely exceeded UK pounds 20000 over an 80-year follow-up, but did for shorter periods. CONCLUSIONS: H. pylori screening may be cost-effective in the long term. However, before screening can be recommended further evidence is needed to resolve some of the uncertainties, particularly over the efficacy of eradication on risk of gastric cancer, the risk associated with complicated peptic ulcers, and the effect of more widespread opportunistic testing of patients with dyspepsia. PMID- 14561270 TI - Review article. Studying climate effects on ecology through the use of climate indices: the North Atlantic Oscillation, El Nino Southern Oscillation and beyond. AB - Whereas the El Nino Southern Oscillation (ENSO) affects weather and climate variability worldwide, the North Atlantic Oscillation (NAO) represents the dominant climate pattern in the North Atlantic region. Both climate systems have been demonstrated to considerably influence ecological processes. Several other large-scale climate patterns also exist. Although less well known outside the field of climatology, these patterns are also likely to be of ecological interest. We provide an overview of these climate patterns within the context of the ecological effects of climate variability. The application of climate indices by definition reduces complex space and time variability into simple measures, 'packages of weather'. The disadvantages of using global climate indices are all related to the fact that another level of problems are added to the ecology climate interface, namely the link between global climate indices and local climate. We identify issues related to: (i) spatial variation; (ii) seasonality; (iii) non-stationarity; (iv) nonlinearity; and (v) lack of correlation in the relationship between global and local climate. The main advantages of using global climate indices are: (i) biological effects may be related more strongly to global indices than to any single local climate variable; (ii) it helps to avoid problems of model selection; (iii) it opens the possibility for ecologists to make predictions; and (iv) they are typically readily available on Internet. PMID- 14561271 TI - Migration route and spawning area fidelity by North Sea plaice. AB - Data from plaice, Pleuronectes platessa L., tagged with electronic data storage tags, were used to test whether these fishes exhibited migration route and spawning area fidelity in successive spawning seasons. Depth and temperature data were recorded for each fish over 365-512 days in the central North Sea and this information was used to reconstruct movements based on tidal locations. We discovered highly directed seasonal migrations from the winter spawning area south of a major topographical feature, Dogger Bank Tail End, to summer feeding grounds 250 km to the north in deep, cold, thermally stratified water. Our results show synchronous timing of migration, repeated pre- and post-spawning migration routes and 100% spawning area fidelity, including two individuals that returned to within 20 km of their previous season's spawning location. This is the first study to provide a complete reconstruction of annual migrations by individual fishes, showing strong homing behaviour along consistent migration routes. PMID- 14561272 TI - Multiple density-dependence mechanisms regulate a migratory bird population during the breeding season. AB - The mechanisms regulating bird populations are poorly understood and controversial. We provide evidence that a migratory songbird, the black-throated blue warbler (Dendroica caerulescens), is regulated by multiple density dependence mechanisms in its breeding quarters. Evidence of regulation includes: stability in population density during 1969-2002, strong density dependence in time-series analyses of this period, an inverse relationship between warbler density and annual fecundity, and a positive relationship between annual fecundity and recruitment of yearlings in the subsequent breeding season. Tests of the mechanisms causing regulation were carried out within the Hubbard Brook Experimental Forest, New Hampshire, during 1997-1999. When individuals from abutting territories were experimentally removed in a homogeneous patch of high quality habitat, the fecundity of focal pairs nearly doubled, revealing a locally operating crowding mechanism. A site-dependence mechanism was indicated by an inverse relationship between population size and mean territory quality, as well as by greater annual fecundity on the sites that were most frequently occupied and of highest quality. These site-dependence relationships were revealed by intensive monitoring of territory quality and demography at the landscape spatial scale. Crowding and site-dependence mechanisms, therefore, acted simultaneously but at different spatial scales to regulate local abundance of this migratory bird population. PMID- 14561273 TI - Climate and density shape population dynamics of a marine top predator. AB - Long-term studies have documented that climate fluctuations affect the dynamics of populations, but the relative influence of stochastic and density-dependent processes is still poorly understood and debated. Most studies have been conducted on terrestrial systems, and lacking are studies on marine systems explicitly integrating the fact that most populations live in seasonal environments and respond to regular or systematic environmental changes. We separated winter from summer mortality in a seabird population, the blue petrel Halobaena caerulea, in the southern Indian Ocean where the El Nino/Southern Oscillation effects occur with a 3-4-year lag. Seventy per cent of the mortality occurred in winter and was linked to climatic factors, being lower during anomalous warm events. The strength of density dependence was affected by climate, with population crashes occurring when poor conditions occurred at high densities. We found that an exceptionally long-lasting warming caused a ca. 40% decline of the population, suggesting that chronic climate change will strongly affect this top predator. These findings demonstrate that populations in marine systems are particularly susceptible to climate variation through complex interactions between seasonal mortality and density-dependent effects. PMID- 14561274 TI - Impacts of marine-derived nutrients on stream ecosystem functioning. AB - Energy and nutrient subsidies transported across ecosystem boundaries are increasingly appreciated as key drivers of consumer-resource dynamics. As purveyors of pulsed marine-derived nutrients (MDN), spawning salmon are one such cross-ecosystem subsidy to freshwaters connected to the north Pacific. We examined how salmon carcasses influenced detrital processing in an oligotrophic stream. Experimental manipulations of MDN inputs revealed that salmon carcasses indirectly reduced detrital processing in streams through temporarily decoupling the detrital resource-consumer relationship, in which detrital consumers shifted their diet to the high-nutrient resource, i.e. salmon carcasses. The average decomposition rate of alder leaves with salmon carcass addition was significantly lower than that without the carcass, which was associated with lower abundance and biomass of detritivorous Trichoptera on the carcass-treated leaves. There were generally larger in size Trichopteran detritivores on the carcasses than on leaves. These results imply that cross-boundary MDN subsidies indirectly retard the ecosystem processing of leaf litter within the short term, but may enhance those food-limited detritivorous consumers. Because unproductive freshwaters in the Pacific northwest are highly dependent upon the organic matter inputs from surrounding forests, this novel finding has implications for determining conservation and management strategies of salmon-related aquatic ecosystems, in terms of salmon habitat protection and fisheries exploitation. PMID- 14561276 TI - Magnetic orientation in birds: non-compass responses under monochromatic light of increased intensity. AB - Migratory Australian silvereyes (Zosterops lateralis) were tested under monochromatic light at wavelengths of 424 nm blue and 565 nm green. At a low light level of 7 x 10(15) quanta m(-2) s(-1) in the local geomagnetic field, the birds preferred their seasonally appropriate southern migratory direction under both wavelengths. Their reversal of headings when the vertical component of the magnetic field was inverted indicated normal use of the avian inclination compass. A higher light intensity of 43 x 10(15) quanta m(-2) s(-1), however, caused a fundamental change in behaviour: under bright blue, the silvereyes showed an axial tendency along the east-west axis; under bright green, they showed a unimodal preference of a west-northwesterly direction that followed a shift in magnetic north, but was not reversed by inverting the vertical component of the magnetic field. Hence it is not based on the inclination compass. The change in behaviour at higher light intensities suggests a complex interaction between at least two receptors. The polar nature of the response under bright green cannot be explained by the current models of light-dependent magnetoreception and will lead to new considerations on these receptive processes. PMID- 14561275 TI - Temporal analysis of archived samples indicates marked genetic changes in declining North Sea cod (Gadus morhua). AB - Despite increasing evidence that current exploitation rates can contribute to shifts in life-history traits and the collapse of marine fish stocks, few empirical studies have investigated the likely evolutionary impacts. Here, we used DNA recovered from a temporal series of archived North Sea cod (Gadus morhua) otoliths, to investigate genetic diversity within the Flamborough Head population between 1954 and 1998, during which time the population underwent two successive declines. Microsatellite data indicated a significant reduction in genetic diversity between 1954 and 1970 (total number of alleles: 1954, 46; 1960, 42; 1970, 37), and a subsequent recovery between 1970 and 1998 (total number of alleles: 1970, 37; 1981, 42; 1998, 45). Furthermore, estimates of genetic differentiation (F(ST) and R(ST)) showed a significant divergence between 1998 and earlier samples. Data are consistent with a period of prolonged genetic drift, accompanied by a replacement of the Flamborough Head population through an increased effective migration rate that occurred during a period of high exploitation and appreciable demographic and phenotypic change. Other studies indicate that diversity at neutral microsatellite loci may be correlated with variability at selected genes, thus compromising a population's subsequent recovery and adaptive potential. Such effects are especially pertinent to North Sea cod, which are threatened by continuing exploitation and rising sea temperatures. PMID- 14561277 TI - Modelling studies on bovine spongiform encephalopathy occurrence to assist in the review of the over 30 months rule in Great Britain. AB - The objective of this study was to contribute to a risk assessment to review the over 30 months (OTM) scheme in cattle, whereby all cattle aged over 30 months slaughtered in the UK are removed from the human food chain. We use back calculation methods to estimate the impact of changes to the OTM rule, by using passive and active surveillance data collected between 1 July 2001 and 30 June 2002. There are two types of change considered: increasing the age limit and allowing animals born after a certain date into the food chain. Results indicate that under the OTM rule less than 1 animal in the last 12 months of the incubation period would enter the food chain in 2003. The birth date changes considered and small changes to the upper age limit would increase this number by a relatively small amount. PMID- 14561278 TI - Oral creatine monohydrate supplementation improves brain performance: a double blind, placebo-controlled, cross-over trial. AB - Creatine supplementation is in widespread use to enhance sports-fitness performance, and has been trialled successfully in the treatment of neurological, neuromuscular and atherosclerotic disease. Creatine plays a pivotal role in brain energy homeostasis, being a temporal and spatial buffer for cytosolic and mitochondrial pools of the cellular energy currency, adenosine triphosphate and its regulator, adenosine diphosphate. In this work, we tested the hypothesis that oral creatine supplementation (5 g d(-1) for six weeks) would enhance intelligence test scores and working memory performance in 45 young adult, vegetarian subjects in a double-blind, placebo-controlled, cross-over design. Creatine supplementation had a significant positive effect (p < 0.0001) on both working memory (backward digit span) and intelligence (Raven's Advanced Progressive Matrices), both tasks that require speed of processing. These findings underline a dynamic and significant role of brain energy capacity in influencing brain performance. PMID- 14561279 TI - Inbreeding depresses immune response in song sparrows (Melospiza melodia): direct and inter-generational effects. AB - A thorough knowledge of relationships between host genotype and immunity to parasitic infection is required to understand parasite-mediated mechanisms of genetic and population change. It has been suggested that immunity may decline with inbreeding. However, the relationship between inbreeding level and a host's response to a novel immune challenge has not been investigated in a natural population. We used the pedigreed population of song sparrows (Melospiza melodia) inhabiting Mandarte Island, Canada, to test the hypothesis that a sparrow's cell mediated immune response (CMI) to an experimental challenge would decline with individual or parental inbreeding. CMI in 6-day-old chicks declined significantly with their mother's coefficient of inbreeding, demonstrating an inter generational effect of maternal inbreeding on offspring immunity. In fledged juveniles and adult sparrows, CMI declined markedly with an individual's own coefficient of inbreeding, but not its mother's. This relationship was consistent across seasons, and was not attributable solely to heterosis in offspring of immigrant breeders. CMI also declined with age and increased with body condition in adult sparrows, but inbreeding explained 37% of the total variation. We emphasize the implications of this dramatic inbreeding depression in cell mediated immunity for theories of parasite-mediated evolution and the susceptibility of small, inbred populations. PMID- 14561280 TI - Nonlinear and correlational sexual selection on 'honest' female ornamentation. AB - Female ornamentation has long been overlooked because of the greater prevalence of elaborate displays in males. However, the circumstances under which females would benefit from honestly signalling their quality are limited. Females are not expected to invest in ornamentation unless the fitness benefits of the ornament exceed those derived from investing the resources directly into offspring. It has been proposed that when females gain direct benefits from mating, females may instead be selected for ornamentation that deceives males about their reproductive state. In the empidid dance flies, males frequently provide nuptial gifts and it is usually only the female that is ornamented. Female traits in empidids, such as abdominal sacs and enlarged pinnate leg scales, have been proposed to 'deceive' males into matings by disguising egg maturity. We quantified sexual selection in the dance fly Rhamphomyia tarsata and found escalating, quadratic selection on pinnate scales and that pinnate scales honestly reflect female fecundity. Mated females had a larger total number and more mature eggs than unmated females, highlighting a potential benefit rather than a cost of male mate choice. We also show correlational selection on female pinnate scales and fecundity. Correlational selection, equivalent investment patterns or increased nutrition from nuptial gifts may all maintain honesty in female ornamentation. PMID- 14561281 TI - Second to fourth digit ratio, testosterone and perceived male dominance. AB - Previous studies have shown that male faces with extreme features associated with testosterone are perceived as dominant and masculine. Women have been reported to prefer more masculinized male faces as they may consider testosterone markers to be an 'honest' indication of good health, and such considerations may underlie their aesthetic preferences. However, pronounced testosterone facial markers are also associated with dominance, and several negative personality traits. This suggests that female aesthetic preferences may be an adaptive compromise between positive attributes associated with higher than average testosterone, and negative attributes associated with more extreme masculinization. This current study attempts to clarify the role of hormone markers in female perceptions of dominance, masculinity and attractiveness, in male facial images. Recent evidence suggests that the relative length of the 2nd to 4th finger (2D : 4D ratio) is a pointer to prenatal testosterone levels and may thus serve as a window to the prenatal hormonal environment. We measured 2D : 4D in a sample of male college students and took salivary samples to analyse circulating levels of testosterone. Women rated facial images of these males for dominance, masculinity and attractiveness. Our results show that male 2D : 4D was significantly negatively related to perceived dominance and masculinity but not attractiveness. Circulating testosterone levels were not related to dominance, masculinity or attractiveness. These findings suggest that: (i) high prenatal levels of testosterone serve to 'organize' male facial features to subsequently reflect dominance and masculine characteristics presumably activated during puberty; and (ii) attractiveness is not directly related to testosterone levels. We conclude that facial dominance and masculinity reflect a male's perceived status rather than his physical attraction to women. PMID- 14561282 TI - Positive force feedback in bouncing gaits? AB - During bouncing gaits (running, hopping, trotting), passive compliant structures (e.g. tendons, ligaments) store and release part of the stride energy. Here, active muscles must provide the required force to withstand the developing tendon strain and to compensate for the inevitable energy losses. This requires an appropriate control of muscle activation. In this study, for hopping, the potential involvement of afferent information from muscle receptors (muscle spindles, Golgi tendon organs) is investigated using a two-segment leg model with one extensor muscle. It is found that: (i) positive feedbacks of muscle-fibre length and muscle force can result in periodic bouncing; (ii) positive force feedback (F+) stabilizes bouncing patterns within a large range of stride energies (maximum hopping height of 16.3 cm, almost twofold higher than the length feedback); and (iii) when employing this reflex scheme, for moderate hopping heights (up to 8.8 cm), an overall elastic leg behaviour is predicted (hopping frequency of 1.4-3 Hz, leg stiffness of 9-27 kN m(-1)). Furthermore, F+ could stabilize running. It is suggested that, during the stance phase of bouncing tasks, the reflex-generated motor control based on feedbacks might be an efficient and reliable alternative to central motor commands. PMID- 14561283 TI - A bacterial symbiont in the Bacteroidetes induces cytoplasmic incompatibility in the parasitoid wasp Encarsia pergandiella. AB - Vertically transmitted symbionts of arthropods have been implicated in several reproductive manipulations of their hosts. These include cytoplasmic incompatibility (CI), parthenogenesis induction in haplodiploid species (PI), feminization and male killing. One symbiont lineage in the alpha-Proteobacteria, Wolbachia, is the only bacterium known to cause all of these effects, and has been thought to be unique in causing CI, in which the fecundity of uninfected females is reduced after mating with infected males. Here, we provide evidence that an undescribed symbiont in the Bacteroidetes group causes CI in a sexual population of the parasitic wasp Encarsia pergandiella. Wasps were crossed in all four possible combinations of infected and uninfected individuals. In the cross predicted to be incompatible, infected (I) males x uninfected (U) females, progeny production was severely reduced, with these females producing only 12.6% of the number of progeny in other crosses. The incompatibility observed in this haplodiploid species was the female mortality type; dissections showed that most progeny from the incompatible cross died as eggs. The 16S rDNA sequence of this symbiont is 99% identical to a parthenogenesis-inducing symbiont in other Encarsia, and 96% identical to a feminizing symbiont in haplodiploid Brevipalpus mites. Thus, this recently discovered symbiont lineage is capable of inducing three of the four principal manipulations of host reproduction known to be caused by Wolbachia. PMID- 14561284 TI - Pulsed mass recruitment by a stingless bee, Trigona hyalinata. AB - Research on bee communication has focused on the ability of the highly social bees, stingless bees (Hymenoptera, Apidae, Meliponini) and honeybees (Apidae, Apini), to communicate food location to nest-mates. Honeybees can communicate food location through the famous waggle dance. Stingless bees are closely related to honeybees and communicate food location through a variety of different mechanisms, many of which are poorly understood. We show that a stingless bee, Trigona hyalinata, uses a pulsed mass-recruitment system that is highly focused in time and space. Foragers produced an ephemeral, polarized, odour trail consisting of mandibular gland secretions. Surprisingly, the odour trail extended only a short distance away from the food source, instead of providing a complete trail between the nest and the food source (as has been described for other stingless bees). This abbreviated trail may represent an intermediate strategy between full-trail marking, found in some stingless bees, and odour marking of the food alone, found in stingless bees and honeybees. PMID- 14561285 TI - Curtailing transmission of severe acute respiratory syndrome within a community and its hospital. AB - Severe acute respiratory syndrome (SARS) has been transmitted extensively within hospitals, and healthcare workers (HCWs) have comprised a large proportion of SARS cases worldwide. We present a stochastic model of a SARS outbreak in a community and its hospital. For a range of basic reproductive numbers (R(0)) corresponding to conditions in different cities (but with emphasis on R(0) approximately 3 as reported for Hong Kong and Singapore), we evaluate contact precautions and case management (quarantine and isolation) as containment measures. Hospital-based contact precautions emerge as the most potent measures, with hospital-wide measures being particularly important if screening of HCWs is inadequate. For R(0) = 3, case isolation alone can control a SARS outbreak only if isolation reduces transmission by at least a factor of four and the mean symptom-onset-to-isolation time is less than 3 days. Delays of a few days in contact tracing and case identification severely degrade the utility of quarantine and isolation, particularly in high-transmission settings. Still more detrimental are delays between the onset of an outbreak and the implementation of control measures; for given control scenarios, our model identifies windows of opportunity beyond which the efficacy of containment efforts is reduced greatly. By considering pathways of transmission in our system, we show that if hospital based transmission is not halted, measures that reduce community-HCW contact are vital to preventing a widespread epidemic. The implications of our results for future emerging pathogens are discussed. PMID- 14561286 TI - Mating in a viscous universe: the race is to the agile, not to the swift. AB - Charles Darwin's theory of natural selection has at its focal point the mating success of organisms. Among male animals, large body size is widely seen as the principal determinant of mating success. However, where mating takes place in a three-dimensional arena such as water, the arboreal habitat or air, small size with its concomitant aerobatic advantages might be advantageous. Despite considerable interest, the relationship between aerobatic ability and mating success has not yet been demonstrated in a single animal species. Here, we test the hypothesis that the known mating success of small male midges is due to their greater aerobatic ability. To do this, male midges collected from leks in the wild were flown and their flight paths in free flight were recorded on high-speed cameras in the laboratory. Four flight parameters that would seem relevant to male mate acquisition in flight, i.e. acceleration, maximum speed, tortuosity and turn-rate, were analysed with respect to body size. We show that, while in terms of maximum speed there was no detectable difference between small and large males, small males outperformed larger ones with respect to acceleration, tortuosity and turn-rate. We conclude that the hypothesis that small males gain their mating advantage through aerobatic superiority is consistent with the observations reported here. PMID- 14561287 TI - Population dynamics of small mammals in semi-arid regions: a comparative study of demographic variability in two rodent species. AB - The seasonally determined demographic structure of two semi-arid rodents, both agricultural pest species (the leaf-eared mouse (Phyllotis darwini) in Chile and the multimammate mouse (Mastomys natalensis) in Tanzania), is analysed using capture-mark-recapture (CMR) statistical models and measures for elasticity (the relative change in the growth rate due to a relative unit change in the parameter of concern) derived from projection linear matrix models. We demonstrate that reproduction and survival during the breeding season contribute approximately equally to population growth in the leaf-eared mouse, whereas the multimammate mouse is characterized by a more clearly defined seasonal structure into breeding and non-breeding seasons and that reproduction contributes far more than survival during the breeding season. On this basis, we discuss evolutionary and applied (pest control) issues. Regarding the evolution of life histories (leading to a maximization of the overall net annual growth rate), we suggest that for the leaf eared mouse, features favouring survival throughout the year will provide selective value, but that during the main breeding season, features favouring reproduction and survival are about equally favourable. For the multimammate mouse, features favouring survival are particularly important outside the breeding season, whereas during the breeding season features favouring reproduction are more important. Regarding pest control (aiming at reducing the overall net annual growth rate), we suggest that (ignoring economic considerations) affecting survival outside the main breeding season is particularly effective for the leaf-eared mouse, a feature that is even more the case for the multimammate mouse. In sum, we demonstrate through this comparative study that much is to be learnt from studying the dynamics of fluctuating small rodents-a focal issue within much of population ecology. PMID- 14561288 TI - Detecting sexually antagonistic coevolution with population crosses. AB - The result of population crosses on traits such as mating rate, oviposition rate and survivorship are increasingly used to distinguish between modes of coevolution between the sexes. Two key hypotheses, erected from a verbal theory of sexually antagonistic coevolution, have been the subject of several recent tests. First, statistical interactions arising in population crosses are suggested to be indicative of a complex signal/receiver system. In the case of oviposition rates, an interaction between populations (x, y and z) would be indicated by the rank order of female oviposition rates achieved by x, y and z males changing depending upon the female (x, y or z) with which they mated. Second, under sexually antagonistic coevolution females will do 'best' when mated with their own males, where best is defined by the weakest response to the signal and the highest fitness. We test these hypotheses by crossing strains generated from a formal model of sexually antagonistic coevolution. Strains differ in the strength of natural selection acting on male and female traits. In our model, we assume sexually antagonistic coevolution of a single male signal and female receptor. The female receptor is treated as a preference function where both the slope and intercept of the function can evolve. Our results suggest that neither prediction is consistently supported. Interactions are not diagnostic of complex signal-receiver systems, and even under sexually antagonistic coevolution, females may do better mating with males of strains other than their own. These results suggest a reinterpretation of several recent experiments and have important implications for developing theories of speciation when sexually antagonistic coevolution is involved. PMID- 14561289 TI - Stochastic elimination of cancer cells. AB - Tissues of multicellular organisms consist of stem cells and differentiated cells. Stem cells divide to produce new stem cells or differentiated cells. Differentiated cells divide to produce new differentiated cells. We show that such a tissue design can reduce the rate of fixation of mutations that increase the net proliferation rate of cells. It has, however, no consequence for the rate of fixation of neutral mutations. We calculate the optimum relative abundance of stem cells that minimizes the rate of generating cancer cells. There is a critical fraction of stem cell divisions that is required for a stochastic elimination ('wash out') of cancer cells. PMID- 14561290 TI - Costs of immune response in cold-stressed laboratory mice selected for high and low basal metabolism rates. AB - To study whether mounting an immune response is energetically costly, mice from two lines divergently selected for high (H-BMR) and low (L-BMR) basal metabolic rate (BMR) were immunized with sheep red blood cells. Their energy budgets were then additionally burdened by sudden transfer from an ambient temperature of 23 degrees C to 5 degrees C. We found that the immune response of H-BMR mice was lower than that of L-BMR mice. However, the interaction between line affiliation and ambient temperature was not significant and cold exposure did not result in immunosuppression in either line. At 23 degrees C the animals of both lines seemed to cover the costs of immune response by increasing food consumption and digestive efficiency. This was not observed at 5 degrees C, so these costs must have been covered at the expense of other components of the energy budget. Cold exposure itself elicited a considerable increase in food intake and the mass of internal organs, which were also heavier in H-BMR than in L-BMR mice. However, irrespective of the temperature or line affiliation, immunized mice had smaller intestines, while cold-exposed immunized mice had smaller hearts. Furthermore, the observed larger mass of the liver and kidneys in immunized mice of both lines kept at 23 degrees C was not observed at 5 degrees C. Hence, immunization compromised upregulation of the function of metabolically active internal organs, essential for meeting the energetic demands of cold. We conclude that the difficulties with a straightforward demonstration of the energetic costs of immune responses in these animals stem from the extreme flexibility of their energy budgets. PMID- 14561291 TI - Kin selection may influence fostering behaviour in Antarctic fur seals (Arctocephalus gazella). AB - Fostering confers obvious advantages to the offspring but is seemingly costly to the caregiver. Such behaviour is particularly paradoxical in seals where the energetic investment in milk is very high and has led to the suggestion that this behaviour may have evolved through either kin selection or reciprocity. We used a combination of genetic and behavioural data to investigate whether kin selection plays a role in the fostering behaviour observed in a well-studied population of Antarctic fur seals (Arctocephalus gazella) from Bird Island, South Georgia. Genotypic data from eight highly polymorphic microsatellite markers were used to estimate relatedness among mother-pup pairs, foster mother-pup pairs and the total population. Mean relatedness was found to be significantly higher for foster mother-pup pairs than that observed for the total population, suggesting that kin selection could have a role in the maintenance of fostering behaviour in this species. PMID- 14561292 TI - Patterns of morphological integration in marine modular organisms: supra-module organization in branching octocoral colonies. AB - Despite the relative simplicity of their modular growth, marine invertebrates such as arborescent gorgonian octocorals (Octocorallia: Cnidaria) generate complex colonial forms. Colony form in these taxa is a consequence of modular (polyp) replication, and if there is a tight integration among modular and supramodular traits (e.g. polyp aperture, inter-polyp spacing, branch thickness, internode and branch length), then changes at the module level may lead to changes in colony architecture. Alternatively, different groups of traits may evolve semi-independently (or conditionally independent). To examine the patterns of integration among morphological traits in Caribbean octocorals, we compared five morphological traits across 21 species, correcting for the effects of phylogenetic relationships among the taxa. Graphical modelling and phylogenetic independence contrasts among the five morphological characters indicate two groups of integrated traits based on whether they were polyp- or colony-level traits. Although all characters exhibited bivariate associations, multivariate analyses (partial correlation coefficients) showed the strongest integration among the colony-level characters (internode distance and branch length). It is a quantitative demonstration that branching characters within the octocorals studied are independent of characters of the polyps. Despite the universally recognized modularity of octocorals at the level of polyps, branching during colony development may represent an emergent level of integration and modularity. PMID- 14561293 TI - Absence of nepotism in the harassment of duelling queens by honeybee workers. AB - Nepotism shapes interactions among the members of almost every animal society. However, clear evidence of nepotism within highly cooperative insect societies, such as ant, wasp and honeybee colonies, is rare. Recent empirical findings suggest that nepotism occurs within honeybee colonies where kin-selection theory most strongly predicts its existence: during the lethal queen-queen duels that determine which of several young queens will become the colony's next queen. In this study, I test whether worker bees act nepotistically by hindering duelling queens that are distantly related to themselves. I accomplished this by observing labelled workers harassing duelling queen bees in observation hives and subsequently by determining worker-queen relatedness using DNA microsatellites. I show that the workers that harassed duelling queens were neither more-closely nor more-distantly related to them than were workers selected randomly from the colony. Thus, workers did not behave nepotistically by hindering half-sister queens more than full-sister queens. These results demonstrate that under certain conditions, natural selection limits the evolution of nepotism within animal societies despite strong theoretical predictions for its existence. PMID- 14561294 TI - Cooler butterflies lay larger eggs: developmental plasticity versus acclimation. AB - We use a full factorial design to investigate the effects of maternal and paternal developmental temperature, as well as female oviposition temperature, on egg size in the butterfly Bicyclus anynana. Butterflies were raised at two different temperatures and mated in four possible sex-by-parental-temperature crosses. The mated females were randomly divided between high and low oviposition temperatures. On the first day after assigning the females to different temperatures, only female developmental temperature affected egg size. Females reared at the lower temperature laid larger eggs than those reared at a higher temperature. When eggs were measured again after an acclimation period of 10 days, egg size was principally determined by the prevailing temperature during oviposition, with females ovipositing at a lower temperature laying larger eggs. In contrast to widely used assumptions, the effects of developmental temperature were largely reversible. Male developmental temperature did not affect egg size in either of the measurements. Overall, developmental plasticity and acclimation in the adult stage resulted in very similar patterns of egg size plasticity. Consequently, we argue that the most important question when testing the significance of acclamatory changes is not at which stage a given plasticity is induced, but rather whether plastic responses to environmental change are adaptive or merely physiological constraints. PMID- 14561295 TI - Paternity analysis reveals opposing selection pressures on crown coloration in the blue tit (Parus caeruleus). AB - In socially monogamous species, extra-pair paternity can increase the variance in reproductive success and thereby the potential for sexual selection on male ornaments. We studied whether male secondary sexual ornaments are selected through within- and/or extra-pair reproductive success in the blue tit (Parus caeruleus). Male blue tits display a bright blue crown plumage, which reflects substantially in the ultraviolet (UV) and previously has been indicated to be an important sexual signal. We show that males with a more UV-shifted crown hue were less cuckolded, which probably resulted from female preference for more ornamented mates. By contrast, however, older males and males with a less UV shifted hue sired more extra-pair young. This probably did not reflect direct female preference, since cuckolders were not less UV-ornamented than the males they cuckolded. Alternatively, a trade-off between UV ornamentation and other traits that enhance extra-pair success could explain this pattern. Our results might reflect two alternative male mating tactics, where more UV-ornamented males maximize within-pair success and less UV-ornamented males maximize extra-pair success. Since crown colour was selected in opposite directions by within-pair and extra-pair paternity, directional selection through extra-pair matings seemed weak, at least in this population and breeding season. Reduced intensity of sexual selection due to alternative mating tactics constitutes a potential mechanism maintaining additive genetic variance of male ornaments. PMID- 14561296 TI - Paternal investment directly affects female reproductive effort in an insect. AB - Female reproductive effort can be influenced by the quality of her mate. In some species, females increase their reproductive effort by differentially allocating resources after mating with high-quality males. Examination of female reproductive effort in relation to male quality has implications for estimating the evolvability of traits and for sexual-selection models. Accurate quantification of reproductive investment is not possible in many species. Butterflies are an exception, as most nectar-feeding species emerge with almost intact reproductive resources, and in some species males provide nutrients at mating that enhance female fecundity. By manipulating male donations and using radioactive isotopes, we quantified the effect of variation in nutrient provisioning on female reproductive effort in two butterfly species. In the greenveined white butterfly, Pieris napi, females increased their reproductive effort after receiving large male donations. By contrast, in the speckled wood, Pararge aegeria, where males do not provide nutrients, female reproductive effort was independent of male ejaculate. Increased reproductive effort in Pieris napi resulted from the production of more eggs, rather than from investing more resources per egg. In this species donating ability is heritable; hence females laying more eggs after mating with high-donating males benefit both through higher fecundity and through the production of high-donating sons. PMID- 14561297 TI - Soay rams target reproductive activity towards promiscuous females' optimal insemination period. AB - Female promiscuity is thought to have resulted in the evolution of male behaviours that confer advantages in the sperm competition that ensues. In mammalian species, males can gain a post-copulatory advantage in this sperm 'raffle' by inseminating females at the optimal time relative to ovulation, leading to the prediction that males should preferentially associate and copulate with females at these times. To the best of our knowledge, we provide the first high-resolution test of this prediction using feral Soay sheep, which have a mating system characterized by male competition for access to highly promiscuous females. We find that competitive males time their mate guarding (and hence copulations) to occur close to the optimal insemination period (OIP), when females are also increasingly likely to 'cooperate' with copulation attempts. Subordinate males practice an alternative mating tactic, where they break the integrity of the consort pair and force copulations on females. The timing of these forced copulations is also targeted towards the OIP. We thus provide quantitative evidence that female promiscuity has resulted in the evolution of reproductive strategies in which males 'load' the sperm raffle by targeting their mating activity towards female OIPs, when the probability of sperm-competition success is at its greatest. PMID- 14561298 TI - Sperm competition and the evolution of gamete morphology in frogs. AB - Despite detailed knowledge of the ultrastructure of spermatozoa, there is a paucity of information on the selective pressures that influence sperm form and function. Theoretical models for both internal and external fertilizers predict that sperm competition could favour the evolution of longer sperm. Empirical tests of the external-fertilization model have been restricted to just one group, the fishes, and these tests have proved equivocal. We investigated how sperm competition affects sperm morphology in externally fertilizing myobatrachid frogs. We also examined selection acting on egg size, and covariation between sperm and egg morphology. Species were ranked according to probability of group spawning and hence risk of sperm competition. Body size, testis size and oviposition environment may also influence gamete traits and were included in our analyses. After controlling for phylogenetic relationships between the species examined, we found that an increased risk of sperm competition was associated with increased sperm head and tail lengths. Path analysis showed that sperm competition had its greatest direct effect on sperm tail length, as might be expected under selection resulting from competitive fertilization. Sperm competition did not influence egg size. Oviposition location had a strong influence on egg size and a weak influence on sperm length, with terrestrial spawners having larger gametes than aquatic spawners. Our analysis revealed significant correlated evolution between egg morphology and sperm morphology. These data provide a conclusive demonstration that sperm competition selects for increased sperm length in frogs, and evidence for evolutionary covariance between aspects of male and female gamete morphology. PMID- 14561299 TI - Matching marine reserve design to reserve objectives. AB - Recent interest in using marine reserves for marine resource management and conservation has largely been driven by the hope that reserves might counteract declines in fish populations and protect the biodiversity of the seas. However, the creation of reserves has led to dissension from some interested groups, such as fishermen, who fear that reserves will do more harm than good. These perceived differences in the effect of marine reserves on various stakeholder interests has led to a contentious debate over their merit. We argue here that recent findings in marine ecology suggest that this debate is largely unnecessary, and that a single general design of a network of reserves of moderate size and variable spacing can meet the needs and goals of most stakeholders interested in marine resources. Given the high fecundity of most marine organisms and recent evidence for limited distance of larval dispersal, it is likely that reserves can both maintain their own biodiversity and service nearby non-reserve areas. In particular, spillover of larger organisms and dispersal of larvae to areas outside reserves can lead to reserves sustaining or even increasing local fisheries. Ultimately, the success of any reserve network requires attention to the uncertainty and variability in dispersal patterns of marine organisms, clear statements of goals by all stakeholder groups and proper evaluation of reserve performance. PMID- 14561300 TI - Consequences of recurrent gene flow from crops to wild relatives. AB - Concern about gene flow from crops to wild relatives has become widespread with the increasing cultivation of transgenic crops. Possible consequences of such gene flow include genetic assimilation, wherein crop genes replace wild ones, and demographic swamping, wherein hybrids are less fertile than their wild parents, and wild populations shrink. Using mathematical models of a wild population recurrently receiving pollen from a genetically fixed crop, we find that the conditions for genetic assimilation are not stringent, and progress towards replacement can be fast, even for disfavoured crop genes. Demographic swamping and genetic drift relax the conditions for genetic assimilation and speed progress towards replacement. Genetic assimilation can involve thresholds and hysteresis, such that a small increase in immigration can lead to fixation of a disfavoured crop gene that had been maintained at a moderate frequency, even if the increase in immigration is cancelled before the gene fixes. Demographic swamping can give rise to 'migrational meltdown', such that a small increase in immigration can lead to not only fixation of a disfavoured crop gene but also drastic shrinkage of the wild population. These findings suggest that the spread of crop genes in wild populations should be monitored more closely. PMID- 14561301 TI - Climate change in Australian tropical rainforests: an impending environmental catastrophe. AB - It is now widely accepted that global climate change is affecting many ecosystems around the globe and that its impact is increasing rapidly. Many studies predict that impacts will consist largely of shifts in latitudinal and altitudinal distributions. However, we demonstrate that the impacts of global climate change in the tropical rainforests of northeastern Australia have the potential to result in many extinctions. We develop bioclimatic models of spatial distribution for the regionally endemic rainforest vertebrates and use these models to predict the effects of climate warming on species distributions. Increasing temperature is predicted to result in significant reduction or complete loss of the core environment of all regionally endemic vertebrates. Extinction rates caused by the complete loss of core environments are likely to be severe, nonlinear, with losses increasing rapidly beyond an increase of 2 degrees C, and compounded by other climate-related impacts. Mountain ecosystems around the world, such as the Australian Wet Tropics bioregion, are very diverse, often with high levels of restricted endemism, and are therefore important areas of biodiversity. The results presented here suggest that these systems are severely threatened by climate change. PMID- 14561302 TI - Telomere variability in the monocotyledonous plant order Asparagales. AB - A group of monocotyledonous plants within the order Asparagales, forming a distinct clade in phylogenetic analyses, was reported previously to lack the 'typical' Arabidopsis-type telomere (TTTAGGG)(n). This stimulated us to determine what has replaced these sequences. Using slot-blot and fluorescent in situ hybridization (FISH) to species within this clade, our results indicate the following. 1. The typical Arabidopsis-type telomeric sequence has been partly or fully replaced by the human-type telomeric sequence (TTAGGG)(n). Species in Allium lack the human-type variant. 2. In most cases the human variant occurs along with a lower abundance of two or more variants of the minisatellite sequences (of seven types evaluated), usually these being the consensus telomeric sequence of Arabidopsis, Bombyx (TTAGG)(n) and Tetrahymena (TTGGGG)(n). FISH shows that the variants can occur mixed together at the telomere. 3. Telomerases generate products with a 6 base pair periodicity and when sequenced they reveal predominantly a reiterated human-type motif. These motifs probably form the 'true telomere' but the error rate of motif synthesis is higher compared with 'typical' plant telomerases. The data indicate that the Asparagales clade is unified by a mutation resulting in a switch from synthesis of Arabidopsis-like telomeres to a low-fidelity synthesis of human-like telomeres. PMID- 14561303 TI - Relative importance of predators and parasitoids for cereal aphid control. AB - Field experiments with manipulations of natural enemies of plant-feeding insects may show how a diverse enemy group ensures an important ecosystem function such as naturally occurring biological pest control. We studied cereal aphid populations in winter wheat under experimentally reduced densities of: (i) ground dwelling generalist predators (mostly spiders, carabid and staphylinid beetles); (ii) flying predators (coccinellid beetles, syrphid flies, gall midges, etc.) and parasitoids (aphidiid wasps), and a combination of (i) and (ii), compared with open controls. Aphid populations were 18% higher at reduced densities of ground dwelling predators, 70% higher when flying predators and parasitoids were removed, and 172% higher on the removal of both enemy groups. Parasitoid wasps probably had the strongest effect, as flying predators occurred only in negligible densities. The great importance of parasitism is a new finding for aphid control in cereal fields. In conclusion, a more detailed knowledge of the mechanisms of natural pest control would help to develop environmentally sound crop management with reduced pesticide applications. PMID- 14561304 TI - Reproductive isolation driven by the combined effects of ecological adaptation and reinforcement. AB - Recent years have seen a resurgence of interest in the process of speciation but few studies have elucidated the mechanisms either driving or constraining the evolution of reproductive isolation. In theory, the direct effects of reinforcing selection for increased mating discrimination where interbreeding produces hybrid offspring with low fitness and the indirect effects of adaptation to different environments can both promote speciation. Conversely, high levels of homogenizing gene flow can counteract the forces of selection. We demonstrate the opposing effects of reinforcing selection and gene flow in Timema cristinae walking-stick insects. The magnitude of female mating discrimination against males from other populations is greatest when migration rates between populations adapted to alternate host plants are high enough to allow the evolution of reinforcement, but low enough to prevent gene flow from eroding adaptive divergence in mate choice. Moreover, reproductive isolation is strongest under the combined effects of reinforcement and adaptation to alternate host plants. Our findings demonstrate the joint effects of reinforcement, ecological adaptation and gene flow on progress towards speciation in the wild. PMID- 14561305 TI - Prevalence of scrapie infection in Great Britain: interpreting the results of the 1997-1998 abattoir survey. AB - An accurate estimate of the prevalence of scrapie infection in the Great Britain (GB) sheep flock is essential when assessing any potential risk to human health through exposure to sheep transmissible spongiform encephalopathies (TSEs). One method for assessing the prevalence is to sample sheep intended for human consumption using a diagnostic test capable of detecting infected animals prior to the onset of clinical signs. An abattoir survey conducted in Great Britain in 1997-1998 tested brain samples from 2809 apparently healthy sheep of which none was found to be positive for scrapie by histopathology or immunohistochemistry (IHC) although 10 were positive for scrapie-associated fibrils (SAF). Subsequently, the tonsils from a subset of the animals sampled were examined using IHC, one of which tested positive. To interpret these results we use a likelihood-based approach, which accounts for the variation in the prevalence of infection with age and test sensitivity and specificity with stage of infection. Combining the results for all of the diagnostic tests yields an estimate of the prevalence of scrapie infection in the GB sheep flock of 0.22% (95% confidence interval: 0.01-0.97%). Moreover, our analysis suggests that all of the diagnostic tests used are very specific (greater than 99%). Indeed, only SAF detection yields a specificity estimate of less than 100%, which helps to account for the high number of samples found to be positive for SAF. PMID- 14561306 TI - Quantifying male attractiveness. AB - Genetic models of sexual selection are concerned with a dynamic process in which female preference and male trait values coevolve. We present a rigorous method for characterizing evolutionary endpoints of this process in phenotypic terms. In our phenotypic characterization the mate-choice strategy of female population members determines how attractive females should find each male, and a population is evolutionarily stable if population members are actually behaving in this way. This provides a justification of phenotypic explanations of sexual selection and the insights into sexual selection that they provide. Furthermore, the phenotypic approach also has enormous advantages over a genetic approach when computing evolutionarily stable mate-choice strategies, especially when strategies are allowed to be complex time-dependent preference rules. For simplicity and clarity our analysis deals with haploid mate-choice genetics and a male trait that is inherited phenotypically, for example by vertical cultural transmission. The method is, however, easily extendible to other cases. An example illustrates that the sexy son phenomenon can occur when there is phenotypic inheritance of the male trait. PMID- 14561307 TI - Superior sperm competitors sire higher-quality young. AB - The evolution of polyandry remains controversial. This is because, unlike males, in many cases multiple mating by females does not increase fecundity and inevitably involves some costs. As a result, a large number of indirect benefit models have been proposed to explain polyandry. One of these, the good sperm hypothesis, posits that high-quality males are better sperm competitors and sire higher-quality offspring. Hence, by mating multiply, females produce offspring of superior quality. Despite being potentially widely applicable across species, this idea has received little attention. In a laboratory experiment with yellow dung flies ( Scathophaga stercoraria ) we found that males that were more successful in sperm competition also had offspring that developed faster. There was no relationship between paternal success in sperm competition and the ability of offspring to survive post-emergence starvation. Since faster development times are likely to be advantageous in this species, our data provide some support for polyandry evolving as a means of producing higher-quality offspring via sperm competition. PMID- 14561308 TI - Variation in restorer genes and primary sexual investment in gynodioecious Plantago coronopus: the trade-off between male and female function. AB - In many gynodioecious species the nuclear inheritance of male fertility is complex and involves multiple (restorer) genes. In addition to restoring plants from the female (male sterile) to the hermaphrodite (male fertile) state, these genes are also thought to play a role in the determination of the quantity of pollen produced by hermaphrodites. The more restorer alleles a hermaphroditic plant possesses, the higher the pollen production. To test this hypothesis I combined the results of crossing studies of the genetics of male sterility with phenotypic data on investment in stamens and ovules among the progeny of plants involved in these studies. The sex ratio (i.e. the frequency of hermaphrodites among the progeny), being a measure of the number of restorer alleles of the maternal plant, was positively related to the investment in pollen (male function), but negatively related to the investment in ovules (female function), in both field and greenhouse experiments. Consequently, a negative correlation between male and female function was observed (trade-off) and it is suggested that antagonistic pleiotropic effects of restorer genes might be the cause. Phenotypic gender, a measure combining investment in both pollen and ovules, was highly repeatable between field and greenhouse, indicating genetic determination of a more male- or female-biased allocation pattern among the studied plants. PMID- 14561309 TI - Molecular systematics of dormice (Rodentia: Gliridae) and the radiation of Graphiurus in Africa. AB - The phylogenetic relationships among the Gliridae (order Rodentia) were assessed using 3430 nucleotides derived from three nuclear fragments (beta-spectrin non erythrocytic 1, thyrotropin and lecithin cholesterol acyl transferase) and one mitochondrial gene (12S rRNA). We included 14 glirid species, representative of seven genera of the three recognized subfamilies (Graphiurinae, Glirinae and Leithiinae) in our analysis. The molecular data identified three evolutionary lineages that broadly correspond to the three extant subfamilies. However, the data suggest that the genus Muscardinus, previously regarded as falling within the Glirinae, should be included in the Leithiinae. Molecular dating using local molecular clocks and partitioned datasets allowed an estimate of the timing of cladogenesis within the glirids. Graphiurus probably diverged early in the group's evolution (40-50 Myr ago) and the three subfamilies diverged contemporaneously, probably in Europe. The radiation within Graphiurus is more recent, with the colonization of Africa by this lineage estimated at ca. 8-10 Myr ago. PMID- 14561310 TI - The ownership signature in mouse scent marks is involatile. AB - Male house mice advertise their territory ownership through urinary scent marks and use individual-specific patterns of major urinary proteins (MUPs) to discriminate between their own scent and that of other males. It is not clear whether recognition occurs through discrimination of the non-volatile proteins or protein-ligand complexes (direct model), or by the detection of volatile ligands that are released from MUPs (indirect model). To examine the mechanism underlying individual scent mark signatures, we compared investigatory and countermarking responses of male laboratory mice presented with male scent marks from a strain with a different MUP pattern, when they could contact the scent or when contact was prevented by a porous nitrocellulose sheet to which proteins bind. Mice investigated scent marks from other males whether these were covered or not, and biochemical analysis confirmed that the porous cover did not prevent the release of volatiles from scent marks. Having gained information through investigation, mice increased their own scent marking only if they had direct contact with another male's urine, failing to do this when contact was prevented. Individual signatures in scent marks thus appear to be carried by non-volatile proteins or by non-volatile protein-ligand complexes, rather than by volatiles emanating from the scent. PMID- 14561311 TI - Precopulatory mate guarding and mating behaviour in the rotifer Epiphanes senta (Monogononta: Rotifera). AB - Epiphanes senta is a littoral rotifer species that occurs in temporary waters and displays a mating behaviour which has not, to my knowledge, so far been described for monogonont rotifers. Monogonont rotifers show distinctive periods within their life cycle during which mictic females appear. Mictic females produce haploid eggs that develop into males or into diapausing eggs if fertilized. The females of E. senta are mostly stationary on the substrate while males are more active swimmers. If they encounter eggs with female embryos of their own species, they attend them and mate with the hatching female. Experiments showed that males are able to discriminate between male, female and diapausing eggs. They exhibit a strong preference for female eggs that are only a few hours away from hatching compared with eggs in early developmental stages. Further experiments did not show any significant differences in male attendance of mictic and amictic eggs. It is hypothesized that males judge the age of a female egg by sensing a chemical that is produced by the growing embryo and diffuses through the egg shell. The male mating behaviour is similar to precopulatory mate guarding known from arthropods but it lacks the monopolization of the female by the male. PMID- 14561312 TI - Steering a virtual blowfly: simulation of visual pursuit. AB - The behavioural repertoire of male flies includes visually guided chasing after moving targets. The visuomotor control system for these pursuits belongs to the fastest found in the animal kingdom. We simulated a virtual fly, to test whether or not experimentally established hypotheses on the underlying control system are sufficient to explain chasing behaviour. Two operating instructions for steering the chasing virtual fly were derived from behavioural experiments: (i) the retinal size of the target controls the fly's forward speed and, thus, indirectly its distance to the target; and (ii) a smooth pursuit system uses the retinal position of the target to regulate the fly's flight direction. Low-pass filters implement neuronal processing time. Treating the virtual fly as a point mass, its kinematics are modelled in consideration of the effects of translatory inertia and air friction. Despite its simplicity, the model shows behaviour similar to that of real flies. Depending on its starting position and orientation as well as on target size and speed, the virtual fly either catches the target or follows it indefinitely without capture. These two behavioural modes of the virtual fly emerge from the control system for flight steering without implementation of an explicit decision maker. PMID- 14561313 TI - The Farm Scale Evaluations of spring-sown genetically modified crops. Introduction. PMID- 14561314 TI - On the rationale and interpretation of the Farm Scale Evaluations of genetically modified herbicide-tolerant crops. AB - Farmland biodiversity and food webs were compared in conventional and genetically modified herbicide-tolerant (GMHT) crops of beet (Beta vulgaris L.), maize (Zea mays L.) and both spring and winter oilseed rape (Brassica napus L.). GMHT and conventional varieties were sown in a split-field experimental design, at 60-70 sites for each crop, spread over three starting years beginning in 2000. This paper provides a background to the study and the rationale for its design and interpretation. It shows how data on environment, field management and the biota are used to assess the current state of the ecosystem, to define the typical arable field and to devise criteria for selecting, sampling and auditing experimental sites in the Farm Scale Evaluations. The main functional and taxonomic groups in the habitat are ranked according to their likely sensitivity to GMHT cropping, and the most responsive target organisms are defined. The value of the seedbank as a baseline and as an indicator of historical trends is proposed. Evidence from experiments during the twentieth century is analysed to show that large changes in field management have affected sensitive groups in the biota by ca. 50% during a year or short run of years--a figure against which to assess any positive or negative effects of GMHT cropping. The analysis leads to a summary of factors that were, and were not, examined in the first 3 years of the study and points to where modelling can be used to extrapolate the effects to the landscape and the agricultural region. PMID- 14561315 TI - Crop management and agronomic context of the Farm Scale Evaluations of genetically modified herbicide-tolerant crops. AB - The Farm Scale Evaluations of genetically modified herbicide-tolerant crops (GMHT) were conducted in the UK from 2000 to 2002 on beet (sugar and fodder), spring oilseed rape and forage maize. The management of the crops studied is described and compared with current conventional commercial practice. The distribution of field sites adequately represented the areas currently growing these crops, and the sample contained sites operated at a range of management intensities, including low intensity. Herbicide inputs were audited, and the active ingredients used and the rates and the timings of applications compared well with current practice for both GMHT and conventional crops. Inputs on sugar beet were lower than, and inputs on spring oilseed rape and forage maize were consistent with, national averages. Regression analysis of herbicide-application strategies and weed emergence showed that inputs applied by farmers increased with weed densities in beet and forage maize. GMHT crops generally received only one herbicide active ingredient per crop, later and fewer herbicide sprays and less active ingredient (for beet and maize) than the conventional treatments. The audit of inputs found no evidence of bias. PMID- 14561317 TI - Weeds in fields with contrasting conventional and genetically modified herbicide tolerant crops. II. Effects on individual species. AB - We compared the effects of the management of genetically modified herbicide tolerant (GMHT) and conventional beet, maize and spring oilseed rape on 12 weed species. We sampled the seedbank before and after cropping. During the season we counted plants and measured seed rain and biomass. Ratios of densities were used to calculate emergence, survival, reproduction and seedbank change. Treatments significantly affected the biomass of six species in beet, eight in maize and five in spring oilseed rape. The effects were generally consistent, with biomass lower in GMHT beet and spring oilseed rape and higher in GMHT maize. With few exceptions, emergence was higher in GMHT crops. Subsequent survival was significantly lowered for eight species in beet and six in spring oilseed rape in the GMHT treatments. It was increased for five species in maize and one in spring oilseed rape. Significant effects on seedbank change were found for four species. However, for many species in beet and spring oilseed rape (19 out of 24 cases), seed densities were lower in the seedbank after GMHT cropping. These differences compounded over time would result in large decreases in population densities of arable weeds. In maize, populations may increase. PMID- 14561316 TI - Weeds in fields with contrasting conventional and genetically modified herbicide tolerant crops. I. Effects on abundance and diversity. AB - We compared the seedbanks, seed rains, plant densities and biomasses of weeds under two contrasting systems of management in beet, maize and spring oilseed rape. Weed seedbank and plant density were measured at the same locations in two subsequent seasons. About 60 fields were sown with each crop. Each field was split, one half being sown with a conventional variety managed according to the farmer's normal practice, the other half being sown with a genetically modified herbicide-tolerant (GMHT) variety, with weeds controlled by a broad-spectrum herbicide. In beet and rape, plant densities shortly after sowing were higher in the GMHT treatment. Following weed control in conventional beet, plant densities were approximately one-fifth of those in GMHT beet. In both beet and rape, this effect was reversed after the first application of broad-spectrum herbicide, so that late-season plant densities were lower in the GMHT treatments. Biomass and seed rain in GMHT crops were between one-third and one-sixth of those in conventional treatments. The effects of differing weed-seed returns in these two crops persisted in the seedbank: densities following the GMHT treatment were about 20% lower than those following the conventional treatment. The effect of growing maize was quite different. Weed density was higher throughout the season in the GMHT treatment. Late-season biomass was 82% higher and seed rain was 87% higher than in the conventional treatment. The difference was not subsequently detectable in the seedbank because the total seed return was low after both treatments. In all three crops, weed diversity was little affected by the treatment, except for transient effects immediately following herbicide application. PMID- 14561318 TI - Invertebrate responses to the management of genetically modified herbicide tolerant and conventional spring crops. I. Soil-surface-active invertebrates. AB - The effects of herbicide management of genetically modified herbicide-tolerant (GMHT) beet, maize and spring oilseed rape on the abundance and diversity of soil surface-active invertebrates were assessed. Most effects did not differ between years, environmental zones or initial seedbanks or between sugar and fodder beet. This suggests that the results may be treated as generally applicable to agricultural situations throughout the UK for these crops. The direction of the effects was evenly balanced between increases and decreases in counts in the GMHT compared with the conventional treatment. Most effects involving a greater capture in the GMHT treatments occurred in maize, whereas most effects involving a smaller capture were in beet and spring oilseed rape. Differences between GMHT and conventional crop herbicide management had a significant effect on the capture of most surface-active invertebrate species and higher taxa tested in at least one crop, and these differences reflected the phenology and ecology of the invertebrates. Counts of carabids that feed on weed seeds were smaller in GMHT beet and spring oilseed rape but larger in GMHT maize. In contrast, collembolan detritivore counts were significantly larger under GMHT crop management. PMID- 14561320 TI - Invertebrates and vegetation of field margins adjacent to crops subject to contrasting herbicide regimes in the Farm Scale Evaluations of genetically modified herbicide-tolerant crops. AB - The effects of management of genetically modified herbicide-tolerant (GMHT) crops on adjacent field margins were assessed for 59 maize, 66 beet and 67 spring oilseed rape sites. Fields were split into halves, one being sown with a GMHT crop and the other with the equivalent conventional non-GMHT crop. Margin vegetation was recorded in three components of the field margins. Most differences were in the tilled area, with fewer smaller effects mirroring them in the verge and boundary. In spring oilseed rape fields, the cover, flowering and seeding of plants were 25%, 44% and 39% lower, respectively, in the GMHT uncropped tilled margins. Similarly, for beet, flowering and seeding were 34% and 39% lower, respectively, in the GMHT margins. For maize, the effect was reversed, with plant cover and flowering 28% and 67% greater, respectively, in the GMHT half. Effects on butterflies mirrored these vegetation effects, with 24% fewer butterflies in margins of GMHT spring oilseed rape. The likely cause is the lower nectar supply in GMHT tilled margins and crop edges. Few large treatment differences were found for bees, gastropods or other invertebrates. Scorching of vegetation by herbicide-spray drift was on average 1.6% on verges beside conventional crops and 3.7% beside GMHT crops, the difference being significant for all three crops. PMID- 14561319 TI - Invertebrate responses to the management of genetically modified herbicide tolerant and conventional spring crops. II. Within-field epigeal and aerial arthropods. AB - The effects of the management of genetically modified herbicide-tolerant (GMHT) crops on the abundances of aerial and epigeal arthropods were assessed in 66 beet, 68 maize and 67 spring oilseed rape sites as part of the Farm Scale Evaluations of GMHT crops. Most higher taxa were insensitive to differences between GMHT and conventional weed management, but significant effects were found on the abundance of at least one group within each taxon studied. Numbers of butterflies in beet and spring oilseed rape and of Heteroptera and bees in beet were smaller under the relevant GMHT crop management, whereas the abundance of Collembola was consistently greater in all GMHT crops. Generally, these effects were specific to each crop type, reflected the phenology and ecology of the arthropod taxa, were indirect and related to herbicide management. These results apply generally to agriculture across Britain, and could be used in mathematical models to predict the possible long-term effects of the widespread adoption of GMHT technology. The results for bees and butterflies relate to foraging preferences and might or might not translate into effects on population densities, depending on whether adoption leads to forage reductions over large areas. These species, and the detritivore Collembola, may be useful indicator species for future studies of GMHT management. PMID- 14561321 TI - Responses of plants and invertebrate trophic groups to contrasting herbicide regimes in the Farm Scale Evaluations of genetically modified herbicide-tolerant crops. AB - Effects of genetically modified herbicide-tolerant (GMHT) and conventional crop management on invertebrate trophic groups (herbivores, detritivores, pollinators, predators and parasitoids) were compared in beet, maize and spring oilseed rape sites throughout the UK. These trophic groups were influenced by season, crop species and GMHT management. Many groups increased twofold to fivefold in abundance between early and late summer, and differed up to 10-fold between crop species. GMHT management superimposed relatively small (less than twofold), but consistent, shifts in plant and insect abundance, the extent and direction of these effects being dependent on the relative efficacies of comparable conventional herbicide regimes. In general, the biomass of weeds was reduced under GMHT management in beet and spring oilseed rape and increased in maize compared with conventional treatments. This change in resource availability had knock-on effects on higher trophic levels except in spring oilseed rape where herbivore resource was greatest. Herbivores, pollinators and natural enemies changed in abundance in the same directions as their resources, and detritivores increased in abundance under GMHT management across all crops. The result of the later herbicide application in GMHT treatments was a shift in resource from the herbivore food web to the detritivore food web. The Farm Scale Evaluations have demonstrated over 3 years and throughout the UK that herbivores, detritivores and many of their predators and parasitoids in arable systems are sensitive to the changes in weed communities that result from the introduction of new herbicide regimes. PMID- 14561323 TI - Scaling properties and symmetrical patterns in the epidemiology of rotavirus infection. AB - The rich epidemiological database of the incidence of rotavirus, as a cause of severe diarrhoea in young children, coupled with knowledge of the natural history of the infection, can make this virus a paradigm for studies of epidemic dynamics. The cyclic recurrence of childhood rotavirus epidemics in unvaccinated populations provides one of the best documented phenomena in population dynamics. This paper makes use of epidemiological data on rotavirus infection in young children admitted to hospital in Melbourne, Australia from 1977 to 2000. Several mathematical methods were used to characterize the overall dynamics of rotavirus infections as a whole and individually as serotypes G1, G2, G3, G4 and G9. These mathematical methods are as follows: seasonal autoregressive integrated moving average (SARIMA) models, power spectral density (PSD), higher-order spectral analysis (HOSA) (bispectrum estimation and quadratic phase coupling (QPC)), detrended fluctuation analysis (DFA), wavelet analysis (WA) and a surrogate data analysis technique. Each of these techniques revealed different dynamic aspects of rotavirus epidemiology. In particular, we confirm the existence of an annual, biannual and a quinquennial period but additionally we found other embedded cycles (e.g. ca. 3 years). There seems to be an overall unique geometric and dynamic structure of the data despite the apparent changes in the dynamics of the last years. The inherent dynamics seems to be conserved regardless of the emergence of new serotypes, the re-emergence of old serotypes or the transient disappearance of a particular serotype. More importantly, the dynamics of all serotypes is multiple synchronized so that they behave as a single entity at the epidemic level. Overall, the whole dynamics follow a scale-free power-law fractal scaling behaviour. We found that there are three different scaling regions in the time-series, suggesting that processes influencing the epidemic dynamics of rotavirus over less than 12 months differ from those that operate between 1 and ca. 3 years, as well as those between 3 and ca. 5 years. To discard the possibility that the observed patterns could be due to artefacts, we applied a surrogate data analysis technique which enabled us to discern if only random components or linear features of the incidence of rotavirus contribute to its dynamics. The global dynamics of the epidemic is portrayed by wavelet-based incidence analysis. The resulting wavelet transform of the incidence of rotavirus crisply reveals a repeating pattern over time that looks similar on many scales (a property called self-similarity). Both the self-similar behaviour and the absence of a single characteristic scale of the power-law fractal-like scaling of the incidence of rotavirus infection imply that there is not a universal inherently more virulent serotype to which severe gastroenteritis can uniquely be ascribed. PMID- 14561322 TI - The functional logic of cortico-pulvinar connections. AB - The pulvinar is an 'associative' thalamic nucleus, meaning that most of its input and output relationships are formed with the cerebral cortex. The function of this circuitry is little understood and its anatomy, though much investigated, is notably recondite. This is because pulvinar connection patterns disrespect the architectural subunits (anterior, medial, lateral and inferior pulvinar nuclei) that have been the traditional reference system. This article presents a simplified, global model of the organization of cortico-pulvinar connections so as to pursue their structure-function relationships. Connections between the cortex and pulvinar are topographically organized, and as a result the pulvinar contains a 'map' of the cortical sheet. However, the topography is very blurred. Hence the pulvinar connection zones of nearby cortical areas overlap, allowing indirect transcortical communication via the pulvinar. A general observation is that indirect cortico-pulvino-cortical circuits tend to mimic direct cortico cortical pathways: this is termed 'the replication principle'. It is equally apt for certain pairs (or groups) of nearby cortical areas that happen not to connect with each other. The 'replication' of this non-connection is achieved by discontinuities and dislocations of the cortical topography within the pulvinar, such that the associated pair of connection zones do not overlap. Certain of these deformations can be used to divide the global cortical topography into specific sub-domains, which form the natural units of a connectional subdivision of the pulvinar. A substantial part of the pulvinar also expresses visual topography, reflecting visual maps in occipital cortex. There are just two well ordered visual maps in the pulvinar, that both receive projections from area V1, and several other occipital areas; the resulting duplication of cortical topography means that each visual map also acts as a separate connection domain. In summary, the model identifies four topographically ordered connection domains, and reconciles the coexistence of visual and cortical maps in two of them. The replication principle operates at and below the level of domain structure. It is argued that cortico-pulvinar circuitry replicates the pattern of cortical circuitry but not its function, playing a more regulatory role instead. Thalamic neurons differ from cortical neurons in their inherent rhythmicity, and the pattern of cortico-thalamic connections must govern the formation of specific resonant circuits. The broad implication is that the pulvinar acts to coordinate cortical information processing by facilitating and sustaining the formation of synchronized trans-areal assemblies; a more pointed suggestion is that, owing to the considerable blurring of cortical topography in the pulvinar, rival cortical assemblies may be in competition to recruit thalamic elements in order to outlast each other in activity. PMID- 14561324 TI - A spherical model for orientation and spatial-frequency tuning in a cortical hypercolumn. AB - A theory is presented of the way in which the hypercolumns in primary visual cortex (V1) are organized to detect important features of visual images, namely local orientation and spatial-frequency. Given the existence in V1 of dual maps for these features, both organized around orientation pinwheels, we constructed a model of a hypercolumn in which orientation and spatial-frequency preferences are represented by the two angular coordinates of a sphere. The two poles of this sphere are taken to correspond, respectively, to high and low spatial-frequency preferences. In Part I of the paper, we use mean-field methods to derive exact solutions for localized activity states on the sphere. We show how cortical amplification through recurrent interactions generates a sharply tuned, contrast invariant population response to both local orientation and local spatial frequency, even in the case of a weakly biased input from the lateral geniculate nucleus (LGN). A major prediction of our model is that this response is non separable with respect to the local orientation and spatial frequency of a stimulus. That is, orientation tuning is weaker around the pinwheels, and there is a shift in spatial-frequency tuning towards that of the closest pinwheel at non-optimal orientations. In Part II of the paper, we demonstrate that a simple feed-forward model of spatial-frequency preference, unlike that for orientation preference, does not generate a faithful representation when amplified by recurrent interactions in V1. We then introduce the idea that cortico-geniculate feedback modulates LGN activity to generate a faithful representation, thus providing a new functional interpretation of the role of this feedback pathway. Using linear filter theory, we show that if the feedback from a cortical cell is taken to be approximately equal to the reciprocal of the corresponding feed forward receptive field (in the two-dimensional Fourier domain), then the mismatch between the feed-forward and cortical frequency representations is eliminated. We therefore predict that cortico-geniculate feedback connections innervate the LGN in a pattern determined by the orientation and spatial frequency biases of feed-forward receptive fields. Finally, we show how recurrent cortical interactions can generate cross-orientation suppression. PMID- 14561325 TI - Inflammation in central nervous system injury. AB - Inflammation is a key component of host defence responses to peripheral inflammation and injury, but it is now also recognized as a major contributor to diverse, acute and chronic central nervous system (CNS) disorders. Expression of inflammatory mediators including complement, adhesion molecules, cyclooxygenase enzymes and their products and cytokines is increased in experimental and clinical neurodegenerative disease, and intervention studies in experimental animals suggest that several of these factors contribute directly to neuronal injury. Most notably, specific cytokines, such as interleukin-1 (IL-1), have been implicated heavily in acute neurodegeneration, such as stroke and head injury. In spite of their diverse presentation, common inflammatory mechanisms may contribute to many neurodegenerative disorders and in some (e.g. multiple sclerosis) inflammatory modulators are in clinical use. Inflammation may have beneficial as well as detrimental actions in the CNS, particularly in repair and recovery. Nevertheless, several anti-inflammatory targets have been identified as putative treatments for CNS disorders, initially in acute conditions, but which may also be appropriate to chronic neurodegenerative conditions. PMID- 14561327 TI - Genetic epidemiology, genetic maps and positional cloning. AB - Genetic epidemiology developed in the middle of the last century, focused on inherited causes of disease but with methods and results applicable to other traits and even forensics. Early success with linkage led to the localization of genes contributing to disease, and ultimately to the Human Genome Project. The discovery of millions of DNA markers has encouraged more efficient positional cloning by linkage disequilibrium (LD), using LD maps and haplotypes in ways that are rapidly evolving. This has led to large international programmes, some promising and others alarming, with laws about DNA patenting and ethical guidelines for responsible research still struggling to be born. PMID- 14561329 TI - Mammal population regulation, keystone processes and ecosystem dynamics. AB - The theory of regulation in animal populations is fundamental to understanding the dynamics of populations, the causes of mortality and how natural selection shapes the life history of species. In mammals, the great range in body size allows us to see how allometric relationships affect the mode of regulation. Resource limitation is the fundamental cause of regulation. Top-down limitation through predators is determined by four factors: (i). body size; (ii). the diversity of predators and prey in the system; (iii). whether prey are resident or migratory; and (iv). the presence of alternative prey for predators. Body size in mammals has two important consequences. First, mammals, particularly large species, can act as keystones that determine the diversity of an ecosystem. I show how keystone processes can, in principle, be measured using the example of the wildebeest in the Serengeti ecosystem. Second, mammals act as ecological landscapers by altering vegetation succession. Mammals alter physical structure, ecological function and species diversity in most terrestrial biomes. In general, there is a close interaction between allometry, population regulation, life history and ecosystem dynamics. These relationships are relevant to applied aspects of conservation and pest management. PMID- 14561328 TI - Human nutrition and food research: opportunities and challenges in the post genomic era. AB - Sequencing of the human genome has opened the door to the most exciting new era for nutritional science. It is now possible to study the underlying mechanisms for diet-health relationships, and in the near future dietary advice (and possibly tailored food products) for promoting optimal health could be provided on an individual basis, in relation to genotype and lifestyle. The role of food in human evolution is briefly reviewed, from palaeolithic times to modern-day hunter-gatherer societies. The aetiology of 'diseases of modern civilization', such as diabetes, heart disease and cancer, and the effect of changes in dietary patterns are discussed. The risk of disease is often associated with common single nucleotide polymorphisms, but the effect is dependent on dietary intake and nutritional status, and is often more apparent in intervention studies employing a metabolic challenge. To understand the link between diet and health, nutritional research must cover a broad range of areas, from molecular to whole body studies, and is an excellent example of integrative biology, requiring a systems biology approach. The annual cost to the National Health Service of diet related diseases is estimated to be in excess of 15 billion, and although diet is a key component of any preventative strategy, it is not given the prominence it deserves. For example, less than 1% of the pound 1.6 billion budget for coronary heart disease is spent on prevention. The polygenic and multifactorial nature of chronic diseases requires substantial resources but the potential rewards, in terms of quality of life and economics, are enormous. It is timely therefore to consider investing in a long-term coordinated national programme for nutrition research, combining nutritional genomics with established approaches, to improve the health of individuals and of the nation. PMID- 14561326 TI - Evolution of photosensory pineal organs in new light: the fate of neuroendocrine photoreceptors. AB - Pineal evolution is envisaged as a gradual transformation of pinealocytes (a gradual regression of pinealocyte sensory capacity within a particular cell line), the so-called sensory cell line of the pineal organ. In most non-mammals the pineal organ is a directly photosensory organ, while the pineal organ of mammals (epiphysis cerebri) is a non-sensory neuroendocrine organ under photoperiod control. The phylogenetic transformation of the pineal organ is reflected in the morphology and physiology of the main parenchymal cell type, the pinealocyte. In anamniotes, pinealocytes with retinal cone photoreceptor-like characteristics predominate, whereas in sauropsids so-called rudimentary photoreceptors predominate. These have well-developed secretory characteristics, and have been interpreted as intermediaries between the anamniote pineal photoreceptors and the mammalian non-sensory pinealocytes. We have re-examined the original studies on which the gradual transformation hypothesis of pineal evolution is based, and found that the evidence for this model of pineal evolution is ambiguous. In the light of recent advances in the understanding of neural development mechanisms, we propose a new hypothesis of pineal evolution, in which the old notion 'gradual regression within the sensory cell line' should be replaced with 'changes in fate restriction within the neural lineage of the pineal field'. PMID- 14561330 TI - Power over reproduction in social hymenoptera. AB - Inclusive fitness theory has been very successful in predicting and explaining much of the observed variation in the reproductive characteristics of insect societies. For example, the theory correctly predicts sex-ratio biasing by workers in relation to the queen's mating frequency. However, within an insect society there are typically multiple reproductive optima, each corresponding to the interest of different individual(s) or parties of interest. When multiple optima occur, which party's interests prevail? Presumably, the interests of the party with the greatest 'power'; the ability to do or act. This article focuses on factors that influence power over colony reproduction. In particular, we seek to identify the principles that may cause different parties of interest to have greater or lesser power. In doing this, we discuss power from two different angles. On the one hand, we discuss general factors based upon non-idiosyncratic biological features (e.g. information, access to and ability to process food) that are likely to be important to all social Hymenoptera. On the other hand, we discuss idiosyncratic factors that depend upon the biology of a taxon at any hierarchical level. We propose that a better understanding of the diversity of reproductive characteristics of insect societies will come from combining inclusive fitness theory with a wide range of other factors that affect relative power in a conflict situation. PMID- 14561331 TI - Resurrecting Van Leeuwenhoek's rotifers: a reappraisal of the role of disaccharides in anhydrobiosis. AB - In 1702, Van Leeuwenhoek was the first to describe the phenomenon of anhydrobiosis in a species of bdelloid rotifer, Philodina roseola. It is the purpose of this review to examine what has been learned since then about the extreme desiccation tolerance in rotifers and how this compares with our understanding of anhydrobiosis in other organisms. Remarkably, much of what is known today about the requirements for successful anhydrobiosis, and the degree of biostability conferred by the dry state, was already determined in principle by the time of Spallanzani in the late 18th century. Most modern research on anhydrobiosis has emphasized the importance of the non-reducing disaccharides trehalose and sucrose, one or other sugar being present at high concentrations during desiccation of anhydrobiotic nematodes, brine shrimp cysts, bakers' yeast, resurrection plants and plant seeds. These sugars are proposed to act as water replacement molecules, and as thermodynamic and kinetic stabilizers of biomolecules and membranes. In apparent contradiction of the prevailing models, recent experiments from our laboratory show that bdelloid rotifers undergo anhydrobiosis without producing trehalose or any analogous molecule. This has prompted us to critically re-examine the association of disaccharides with anhydrobiosis in the literature. Surprisingly, current hypotheses are based almost entirely on in vitro data: there is very limited information which is more than simply correlative in the literature on living systems. In many species, disaccharide accumulation occurs at approximately the same time as desiccation tolerance is acquired. However, several studies indicate that these sugars are not sufficient for anhydrobiosis; furthermore, there is no conclusive evidence, through mutagenesis or functional knockout experiments, for example, that sugars are necessary for anhydrobiosis. Indeed, some plant seeds and micro-organisms, like the rotifer, exhibit excellent desiccation tolerance in the absence of high intracellular sugar concentrations. Accordingly, it seems appropriate to call for a re-evaluation of our understanding of anhydrobiosis and to embark on new experimental programmes to determine the key molecular mechanisms involved. PMID- 14561334 TI - Mathematical modelling of the cellular mechanics of plants. AB - The complex mechanical behaviour of plant tissues reflects the complexity of their structure and material properties. Modelling has been widely used in studies of how cell walls, single cells and tissue respond to loading, both externally applied loading and loads on the cell wall resulting from changes in the pressure within fluid-filled cells. This paper reviews what approaches have been taken to modelling and simulation of cell wall, cell and tissue mechanics, and to what extent models have been successful in predicting mechanical behaviour. Advances in understanding of cell wall ultrastructure and the control of cell growth present opportunities for modelling to clarify how growth-related mechanical properties arise from wall polymeric structure and biochemistry. PMID- 14561333 TI - Modelling approaches in biomechanics. AB - Conceptual, physical and mathematical models have all proved useful in biomechanics. Conceptual models, which have been used only occasionally, clarify a point without having to be constructed physically or analysed mathematically. Some physical models are designed to demonstrate a proposed mechanism, for example the folding mechanisms of insect wings. Others have been used to check the conclusions of mathematical modelling. However, others facilitate observations that would be difficult to make on real organisms, for example on the flow of air around the wings of small insects. Mathematical models have been used more often than physical ones. Some of them are predictive, designed for example to calculate the effects of anatomical changes on jumping performance, or the pattern of flow in a 3D assembly of semicircular canals. Others seek an optimum, for example the best possible technique for a high jump. A few have been used in inverse optimization studies, which search for variables that are optimized by observed patterns of behaviour. Mathematical models range from the extreme simplicity of some models of walking and running, to the complexity of models that represent numerous body segments and muscles, or elaborate bone shapes. The simpler the model, the clearer it is which of its features is essential to the calculated effect. PMID- 14561335 TI - Aspects of skeletal muscle modelling. AB - The modelling of skeletal muscle raises a number of philosophical questions, particularly in the realm of the relationship between different possible levels of representation and explanation. After a brief incursion into this area, a list of desiderata is proposed as a guiding principle for the construction of a viable model, including: comprehensiveness, soundness, experimental consistency, predictive ability and refinability. Each of these principles is illustrated by means of simple examples. The presence of internal constraints, such as incompressibility, may lead to counterintuitive results. A one-panel example is exploited to advocate the use of the principle of virtual work as the ideal tool to deal with these situations. The question of stability in the descending limb of the force-length relation is addressed and a purely mechanical analogue is suggested. New experimental results confirm the assumption that fibre stiffness is positive even in the descending limb. The indeterminacy of the force-sharing problem is traditionally resolved by optimizing a, presumably, physically meaningful target function. After presenting some new results in this area, based on a separation theorem, it is suggested that a more fundamental approach to the problem is the abandoning of optimization criteria in favour of an explicit implementation of activation criteria. PMID- 14561336 TI - Finite element modelling of contracting skeletal muscle. AB - To describe the mechanical behaviour of biological tissues and transport processes in biological tissues, conservation laws such as conservation of mass, momentum and energy play a central role. Mathematically these are cast into the form of partial differential equations. Because of nonlinear material behaviour, inhomogeneous properties and usually a complex geometry, it is impossible to find closed-form analytical solutions for these sets of equations. The objective of the finite element method is to find approximate solutions for these problems. The concepts of the finite element method are explained on a finite element continuum model of skeletal muscle. In this case, the momentum equations have to be solved with an extra constraint, because the material behaves as nearly incompressible. The material behaviour consists of a highly nonlinear passive part and an active part. The latter is described with a two-state Huxley model. This means that an extra nonlinear partial differential equation has to be solved. The problems and solutions involved with this procedure are explained. The model is used to describe the mechanical behaviour of a tibialis anterior of a rat. The results have been compared with experimentally determined strains at the surface of the muscle. Qualitatively there is good agreement between measured and calculated strains, but the measured strains were higher. PMID- 14561337 TI - Modelling cartilage mechanobiology. AB - The growth, maintenance and ossification of cartilage are fundamental to skeletal development and are regulated throughout life by the mechanical cues that are imposed by physical activities. Finite element computer analyses have been used to study the role of local tissue mechanics on endochondral ossification patterns, skeletal morphology and articular cartilage thickness distributions. Using single-phase continuum material representations of cartilage, the results have indicated that local intermittent hydrostatic pressure promotes cartilage maintenance. Cyclic tensile strains (or shear), however, promote cartilage growth and ossification. Because single-phase material models cannot capture fluid exudation in articular cartilage, poroelastic (or biphasic) solid/fluid models are often implemented to study joint mechanics. In the middle and deep layers of articular cartilage where poroelastic analyses predict little fluid exudation, the cartilage phenotype is maintained by cyclic fluid pressure (consistent with the single-phase theory). In superficial articular layers the chondrocytes are exposed to tangential tensile strain in addition to the high fluid pressure. Furthermore, there is fluid exudation and matrix consolidation, leading to cell 'flattening'. As a result, the superficial layer assumes an altered, more fibrous phenotype. These computer model predictions of cartilage mechanobiology are consistent with results of in vitro cell and tissue and molecular biology experiments. PMID- 14561338 TI - Modelling primary and secondary growth processes in plants: a summary of the methodology and new data from an early lignophyte. AB - A mathematical method, based on polar coordinates that allow modelling of primary and secondary growth processes in stems of extant and fossil plants, is summarized and its potential is discussed in comparison with numerical methods using digitizing tablets or electronic image analysing systems. As an example, the modelling of tissue distribution in the internode of an extant sphenopsid (Equisetum hyemale) is presented. In the second half of the paper we present new data of a functional analysis of stem structure and biomechanics of the early lignophyte Tetraxylopteris schmidtii (Middle Devonian) using the polar coordinate method for modelling the tissue distribution in stems of different ontogenetic age. Calculations of the mechanical properties of the stems, based on the modelling of the tissue arrangement, indicate that there is no increase in structural bending modulus throughout the entire development of the plant. The oldest ontogenetic stage has a significantly smaller bending elastic modulus than the intermediate ontogenetic stage, a 'mechanical signal', which is not consistent with a self-supporting growth form. These results, and the ontogenetic variations of the contributions of different stem tissues to the flexural stiffness of the entire stem, are discussed in the evolutionary context of cambial secondary growth. PMID- 14561339 TI - Oscillations of plants' stems and their damping: theory and experimentation. AB - Free oscillations of upright plants' stems, or in technical terms slender tapered rods with one end free, can be modelled by considering the equilibrium between bending moments and moments resulting from inertia. For stems with apical loads and negligible mass of the stem and for stems with finite mass but without top loading, analytical solutions of the differential equations with appropriate boundary conditions are available for a finite number of cases. For other cases approximations leading to an upper and a lower estimate of the frequency of oscillation omega can be derived. For the limiting case of omega = 0, the differential equations are identical with Greenhill's equations for the stability against Euler buckling of slender poles. To illustrate, the oscillation frequencies of 25 spruce trees (Picea sitchensis (Bong.) Carr.) were compared with those calculated on the basis of their morphology, their density and their static elasticity modulus. For Arundo donax L. and Cyperus alternifolius L. the observed oscillation frequency was used in turn to calculate the dynamic elasticity modulus, which was compared with that determined in three-point bending. Oscillation damping was observed for A. donax and C. alternifolius for plants' stems with and without leaves or inflorescence. In C. alternifolius the difference can be attributed to the aerodynamic resistance of the leaves, whereas in A. donax structural damping in addition plays a major role. PMID- 14561340 TI - Inverse and forward dynamics: models of multi-body systems. AB - Connected multi-body systems exhibit notoriously complex behaviour when driven by external and internal forces and torques. The problem of reconstructing the internal forces and/or torques from the movements and known external forces is called the 'inverse dynamics problem', whereas calculating motion from known internal forces and/or torques and resulting reaction forces is called the 'forward dynamics problem'. When stepping forward to cross the street, people use muscle forces that generate angular accelerations of their body segments and, by virtue of reaction forces from the street, a forward acceleration of the centre of mass of their body. Inverse dynamics calculations applied to a set of motion data from such an event can teach us how temporal patterns of joint torques were responsible for the observed motion. In forward dynamics calculations we may attempt to create motion from such temporal patterns, which is extremely difficult, because of the complex mechanical linkage along the chains forming the multi-body system. To understand, predict and sometimes control multi-body systems, we may want to have mathematical expressions for them. The Newton-Euler, Lagrangian and Featherstone approaches have their advantages and disadvantages. The simulation of collisions and the inclusion of muscle forces or other internal forces are discussed. Also, the possibility to perform a mixed inverse and forward dynamics calculation are dealt with. The use and limitations of these approaches form the conclusion. PMID- 14561341 TI - Simple and complex models for studying muscle function in walking. AB - While simple models can be helpful in identifying basic features of muscle function, more complex models are needed to discern the functional roles of specific muscles in movement. In this paper, two very different models of walking, one simple and one complex, are used to study how muscle forces, gravitational forces and centrifugal forces (i.e. forces arising from motion of the joints) combine to produce the pattern of force exerted on the ground. Both the simple model and the complex one predict that muscles contribute significantly to the ground force pattern generated in walking; indeed, both models show that muscle action is responsible for the appearance of the two peaks in the vertical force. The simple model, an inverted double pendulum, suggests further that the first and second peaks are due to net extensor muscle moments exerted about the knee and ankle, respectively. Analyses based on a much more complex, muscle-actuated simulation of walking are in general agreement with these results; however, the more detailed model also reveals that both the hip extensor and hip abductor muscles contribute significantly to vertical motion of the centre of mass, and therefore to the appearance of the first peak in the vertical ground force, in early single-leg stance. This discrepancy in the model predictions is most probably explained by the difference in model complexity. First, movements of the upper body in the sagittal plane are not represented properly in the double-pendulum model, which may explain the anomalous result obtained for the contribution of a hip-extensor torque to the vertical ground force. Second, the double-pendulum model incorporates only three of the six major elements of walking, whereas the complex model is fully 3D and incorporates all six gait determinants. In particular, pelvic list occurs primarily in the frontal plane, so there is the potential for this mechanism to contribute significantly to the vertical ground force, especially during early single-leg stance when the hip abductors are activated with considerable force. PMID- 14561342 TI - Modelling the control of interceptive actions. AB - In recent years, several phenomenological dynamical models have been formulated that describe how perceptual variables are incorporated in the control of motor variables. We call these short-route models as they do not address how perception action patterns might be constrained by the dynamical properties of the sensory, neural and musculoskeletal subsystems of the human action system. As an alternative, we advocate a long-route modelling approach in which the dynamics of these subsystems are explicitly addressed and integrated to reproduce interceptive actions. The approach is exemplified through a discussion of a recently developed model for interceptive actions consisting of a neural network architecture for the online generation of motor outflow commands, based on time to-contact information and information about the relative positions and velocities of hand and ball. This network is shown to be consistent with both behavioural and neurophysiological data. Finally, some problems are discussed with regard to the question of how the motor outflow commands (i.e. the intended movement) might be modulated in view of the musculoskeletal dynamics. PMID- 14561343 TI - Bipedalism in lizards: whole-body modelling reveals a possible spandrel. AB - This paper illustrates how simple mechanical models based on morphological, ethological, ecological and phylogenetic data can add to discussions in evolutionary biology. Bipedal locomotion has evolved on numerous occasions in lizards. Traits that appear repeatedly in independent evolutionary lines are often considered adaptive, but the exact advantages of bipedal locomotion in lizards remain debated. Earlier claims that bipedalism would increase maximal running speed or would be energetically advantageous have been questioned. Here, we use 'whole body' mechanical modelling to provide an alternative solution to the riddle. The starting point is the intermittent running style combined with the need for a high manoeuvrability characterizing many small lizard species. Manoeuvrability benefits from a caudal shift of the centre of mass of the body (body-COM), because forces to change the heading and to align the body to this new heading do not conflict with each other. The caudally situated body-COM, however, might result in a lift of the front part of the body when accelerating (intermittent style), thus resulting in bipedal running bouts. Based on a momentum-impulse approach the effect of acceleration is quantified for a mechanical model, a virtual lizard (three segments) based on the morphometrics of Acanthodactylus erythrurus (a small lacertid lizard). Biologically relevant input (dimensions, inertial properties, step cycle information, etc.) results in an important lift of the front part of the body and observable distances passively covered bipedally as a consequence of the acceleration. In this way, no functional explanation of the phenomenon of lizard bipedalism is required and bipedalism can probably be considered non-adaptive in many cases. This does not exclude, however, some species that may have turned this consequence to their benefit. For instance, instantaneous manipulation of the position of the centre of the body-COM allows stable, persisting bipedal running. Once this was achieved, the bipedal spandrel could be exploited further. PMID- 14561344 TI - Cyberkelp: an integrative approach to the modelling of flexible organisms. AB - Biomechanical models come in a variety of forms: conceptual models; physical models; and mathematical models (both of the sort written down on paper and the sort carried out on computers). There are model structures (such as insect flight muscle and the tendons of rats' tails), model organisms (such as the flying insect, Manduca sexta), even model systems of organisms (such as the communities that live on wave-swept rocky shores). These different types of models are typically employed separately, but their value often can be enhanced if their insights are integrated. In this brief report we explore a particular example of such integration among models, as applied to flexible marine algae. A conceptual model serves as a template for the construction of a mathematical model of a model species of giant kelp, and the validity of this numerical model is tested using physical models. The validated mathematical model is then used in conjunction with a computer-controlled tensile testing apparatus to simulate the loading regime placed on algal materials. The resulting information can be used to create a more precise mathematical model. PMID- 14561345 TI - Dynamics of filaments: modelling the dynamics of driven microfilaments. AB - We describe a method for simulating the inertialess dynamics of a flexible filament immersed in a fluid. Typically, this regime is appropriate for filaments a few micrometres or less in size (flagella that propel micro-organisms for example). We apply the model to two systems; a filament that is wiggled at one end and planar swimming motion characteristic of simple spermatozoa. For the former we find qualitative agreement with theory. The shape is determined by a balance between bending and viscous forces and there is an optimal balance that maximizes the propulsion generated by this mechanism. Quantitatively we find less satisfactory agreement. For the spermatozoa, assuming a relatively naive bending mechanism in the form of a travelling force quadrupole wave, the model generates waveforms in very good agreement with experiment. This is only true, however, if the bending forces acting on the filament are large compared with the viscous forces. Experimental measurements of the tail stiffness imply this should not be the case. We discuss the implications of this observation in the context of the sperm's swimming mechanism. PMID- 14561346 TI - Simulation and analysis of flow patterns around the scleractinian coral Madracis mirabilis (Duchassaing and Michelotti). AB - Three-dimensional morphologies of Madracis mirabilis were obtained using X-ray computed tomography scanning techniques. The morphologies were used to simulate the flow patterns around the colony. In the simulations, the thin-branching low flow morph with a relatively larger branch-spacing was compared with the more compact high-flow morph of M. mirabilis. For both morphologies, the inside-colony flow velocities were computed for Reynolds numbers ranging from 154 to 3840. In the high-flow morph, it was found that in the range of investigated Reynolds numbers a stagnant region develops within the colony, whereas in the low-flow morph the stagnant region disappeared. Experiments done under natural conditions suggest that a morph is adapted to a certain external flow velocity and develops a stagnant region below a particular threshold for the external flow velocity. When the external flow velocity exceeds a certain threshold, which is characteristic for the growth form, the core velocity becomes equal to the external velocity. A potential application of a profile of core velocities for a range of Reynolds numbers for a certain morph is the prediction of the optimal external flow velocity for a certain morph, and this can be used to assess the state of the physical (palaeo-) environment. PMID- 14561347 TI - How to perform measurements in a hovering animal's wake: physical modelling of the vortex wake of the hawkmoth, Manduca sexta. AB - The vortex wake structure of the hawkmoth, Manduca sexta, was investigated using a vortex ring generator. Based on existing kinematic and morphological data, a piston and tube apparatus was constructed to produce circular vortex rings with the same size and disc loading as a hovering hawkmoth. Results show that the artificial rings were initially laminar, but developed turbulence owing to azimuthal wave instability. The initial impulse and circulation were accurately estimated for laminar rings using particle image velocimetry; after the transition to turbulence, initial circulation was generally underestimated. The underestimate for turbulent rings can be corrected if the transition time and velocity profile are accurately known, but this correction will not be feasible for experiments on real animals. It is therefore crucial that the circulation and impulse be estimated while the wake vortices are still laminar. The scaling of the ring Reynolds number suggests that flying animals of about the size of hawkmoths may be the largest animals whose wakes stay laminar for long enough to perform such measurements during hovering. Thus, at low advance ratios, they may be the largest animals for which wake circulation and impulse can be accurately measured. PMID- 14561348 TI - Comparing fluid mechanics models with experimental data. AB - The art of modelling the physical world lies in the appropriate simplification and abstraction of the complete problem. In fluid mechanics, the Navier-Stokes equations provide a model that is valid under most circumstances germane to animal locomotion, but the complexity of solutions provides strong incentive for the development of further, more simplified practical models. When the flow organizes itself so that all shearing motions are collected into localized patches, then various mathematical vortex models have been very successful in predicting and furthering the physical understanding of many flows, particularly in aerodynamics. Experimental models have the significant added convenience that the fluid mechanics can be generated by a real fluid, not a model, provided the appropriate dimensionless groups have similar values. Then, analogous problems can be encountered in making intelligible but independent descriptions of the experimental results. Finally, model predictions and experimental results may be compared if, and only if, numerical estimates of the likely variations in the tested quantities are provided. Examples from recent experimental measurements of wakes behind a fixed wing and behind a bird in free flight are used to illustrate these principles. PMID- 14561349 TI - Approaches to the structural modelling of insect wings. AB - Insect wings lack internal muscles, and the orderly, necessary deformations which they undergo in flight and folding are in part remotely controlled, in part encoded in their structure. This factor is crucial in understanding their complex, extremely varied morphology. Models have proved particularly useful in clarifying the facilitation and control of wing deformation. Their development has followed a logical sequence from conceptual models through physical and simple analytical to numerical models. All have value provided their limitations are realized and constant comparisons made with the properties and mechanical behaviour of real wings. Numerical modelling by the finite element method is by far the most time-consuming approach, but has real potential in analysing the adaptive significance of structural details and interpreting evolutionary trends. Published examples are used to review the strengths and weaknesses of each category of model, and a summary is given of new work using finite element modelling to investigate the vibration properties and response to impact of hawkmoth wings. PMID- 14561350 TI - Physical modelling in biomechanics. AB - Physical models, like mathematical models, are useful tools in biomechanical research. Physical models enable investigators to explore parameter space in a way that is not possible using a comparative approach with living organisms: parameters can be varied one at a time to measure the performance consequences of each, while values and combinations not found in nature can be tested. Experiments using physical models in the laboratory or field can circumvent problems posed by uncooperative or endangered organisms. Physical models also permit some aspects of the biomechanical performance of extinct organisms to be measured. Use of properly scaled physical models allows detailed physical measurements to be made for organisms that are too small or fast to be easily studied directly. The process of physical modelling and the advantages and limitations of this approach are illustrated using examples from our research on hydrodynamic forces on sessile organisms, mechanics of hydraulic skeletons, food capture by zooplankton and odour interception by olfactory antennules. PMID- 14561351 TI - Biomimetic modelling. AB - Biomimetics is seen as a path from biology to engineering. The only path from engineering to biology in current use is the application of engineering concepts and models to biological systems. However, there is another pathway: the verification of biological mechanisms by manufacture, leading to an iterative process between biology and engineering in which the new understanding that the engineering implementation of a biological system can bring is fed back into biology, allowing a more complete and certain understanding and the possibility of further revelations for application in engineering. This is a pathway as yet unformalized, and one that offers the possibility that engineers can also be scientists. PMID- 14561352 TI - Graham Fraser Memorial Lecture 2002. From frogs' legs to pieds-noirs and beyond: some aspects of cochlear implantation. AB - The 2002 Graham Fraser Memorial Lecture deals first with the French origins of cochlear implantation in Paris in the 1950s and the role of Andre Djourno and Charles Eyries. Following this work in Paris Dr William House in Los Angeles continued work on cochlear implants and, subsequently, experimental implant programmes were started in California, Paris, Vienna and Melbourne. The next section of this lecture covers the experimental work of Galvani in establishing the role of electricity in physiology. The results of his first experiments were published in 1791, the year that Mondini produced the first account of a cochlear malformation in a congenitally deaf child. At around the same time sign language for congenitally deaf children was being developed for the first time in Paris by Epee and the first disputes occurred between oralists and those who promoted signing for the education of congenitally deaf children. In a present day cochlear implant programme good results from implanting congenitally deaf children at an early age and implanting adults who have become profoundly deaf are now taken for granted. We do have much to learn, however, from more complex implant candidates and some examples of such candidates are presented. Lastly, looking to the future, the use of PET scanning to try and gain information about how the brain handles the information provided to it by a cochlear implant is described. PMID- 14561353 TI - Nasal snuff: historical review and health related aspects. AB - With cigarette smoking declining in the modern world, the tobacco industry has to look for other products that can keep the old customers and attract new ones. Different forms of smokeless tobacco are currently massively promoted and are gaining in importance. Dry nasal snuff--the oldest known form of tobacco in Europe--is one of them. The health risks associated with it are different to those attributed to smoking and oral wet snuff. The nicotine contained leads to dependency. Its resorption rate is similar to that of smoking, so it could be seen as an adequate substitutional therapy. The risk for cardiovascular diseases is lower, compared to that for smokers. Chronic abuse leads to morphological and functional changes in the nasal mucosa. Although it contains substances that are potentially carcinogenic, at present, there is no firm evidence, relating the use of nasal snuff to a higher incidence of head and neck or other malignancies. PMID- 14561354 TI - High resolution computed tomography and magnetic resonance imaging in the pre operative assessment of cochlear implant patients. AB - Optimal imaging protocols for cochlear implantation have yet to be determined. Pre-operative computed tomography (CT) and magnetic resonance image (MRI) scans are used to assess cochlear anatomy and patency, to delineate surgical access, and to aid in choice of side for implantation. However, opinion still differs as to which modality provides more information in pre-operative assessment, or if, indeed, a combination of the two is superior. The first 88 patients on the Irish National Cochlear Implant Programme (NCIP) were retrospectively studied to determine the accuracy of pre-operative CT and MRI in predicting abnormalities at the time of surgery. Correlation with surgical findings was determined in three separate groups of patients (those who had CT only, those who had MRI only, and those who had both CT and MRI performed). Of the 24 patients that had both CT and MRI performed, both modalities had a 79 per cent correlation with surgical findings. CT and MRI reports concurred in 75 per cent of cases. Specificity and negative predictive value were high (86 per cent and 90 per cent, respectively). CT alone (47 cases) correlated with surgery in 39 cases (83 per cent); MRI alone (17 cases) correlated in 15 cases (88 per cent). The findings of this study suggest that CT and MRI are effective at predicting normal inner ear anatomy, and thus at predicting the patient and the cochlea most suitable for implantation. Both modalities are useful in determining the side of implantation, thus avoiding potential surgical difficulties in cases of unilateral abnormalities. There was no significant difference between the ability of MRI and CT to detect abnormalities at the time of surgery. In this series the combination of CT and MRI has not been shown to be superior to either modality used alone, although anecdotal evidence to the contrary was noted. PMID- 14561355 TI - Methodology for studying the effects of topically applied ear drops on otoacoustic emissions in guinea pigs. AB - Testing of otoacoustic emissions (OAE) has been successfully applied to study outer hair cell function. We used this methodology to evaluate and compare the effect of topically applied normal saline, and gentamicin- and ciprofloxacin containing drops into the middle ear of guinea pigs. All three solutions similarly decreased the distortion product OAE (DPOAE) and all three caused an inflammatory reaction in the guinea pig's middle-ear. OAE failed to detect adequately the ototoxicity of topically administered drops in this animal model because the opening of the bulla affected DPOAE, and both inflamed middle-ear mucosa and osteoneogenesis caused a decrease in DPOAE amplitude. This is the first report examining the influence of topically used otic drops on inner ear function as demonstrated by OAE. PMID- 14561356 TI - Effect of topically applied povidone-iodine on transient evoked otoacoustic emissions in guinea pigs. AB - The possible cochlear toxicity of topically applied povidone-iodine solution was investigated in guinea pigs by measuring transiently evoked otoacoustic emissions (TEOAEs). Seven adult pigmented guinea pigs (14 ears) with a normal acoustic pinna reflex were used. After the baseline TEOAE measurements, 0.1 mL of povidone iodine solution was applied transtympanically into the middle ear in the study group. Saline solution and gentamicin were used as negative and positive control, respectively. TEOAE measurements were repeated on the 10th day, and four weeks after the first application. Tympanometric measurements were also carried out for both ears of all animals at the end of the study. Responses in all ears, which had povidone-iodine applied to them, disappeared in a way similar to those recorded in gentamicin-treated ears. Tympanometric measurements at the end of the study showed 'type A' tympanograms in all ears of all animals. The present study showed that povidone-iodine could have a cochleotoxic effect in guinea pigs. A study in humans with TEOAE before and after middle-ear surgery, where povidone iodine can reach into the middle-ear cavity, may be useful for evaluating this result for clinical practice. PMID- 14561357 TI - Increased post-operative haemorrhage seen in adult coblation tonsillectomy. AB - Coblation is a new soft tissue surgical technique that is being used for tonsillectomy. Published results show a significant decrease in the amount of post-operative pain experienced by patients undergoing coblation tonsillectomy. There has been no published work to date on the incidence of post-operative haemorrhage. From August 2001 to November 2002 one surgeon performed 36 coblation tonsillectomies on adults. On another list he performed 29 by his standard method of dissection and bipolar coagulation. Retrospective analysis found a significant increase in the secondary haemorrhage rate in adult patients undergoing coblation tonsillectomy (22.2 vs. 3.4 per cent). At our department coblation tonsillectomy has been abandoned until further work into its safety has been published. PMID- 14561358 TI - Does synchronous nasal surgery increase the risk of post-operative haemorrhage in adult patients undergoing tonsillectomy? AB - Otolaryngologists occasionally perform simultaneous procedures on their patients, the indication being co-existent pathology. This is not a universally accepted practice as some feel that the post-operative morbidity is increased in such patients. Tonsillectomy is a common otolaryngological procedure and the main life threatening complication, haemorrhage is easily identifiable. This study compares the incidence of post-operative haemorrhage requiring admission in patients undergoing tonsillectomy alone with that in patients undergoing tonsillectomy and nasal surgery. A retrospective study was undertaken of all adult patients undergoing tonsillectomy alone or tonsillectomy and synchronous nasal surgery in Aberdeen Royal Infirmary. Seventy-one patients (34 male) with a mean age of 23 years underwent tonsillectomy and synchronous nasal surgery over a nine-year period 1991-1999. The commonest nasal procedure was reduction of the inferior turbinates (in 48 cases). Three hundred and ninety-eight patients (131 male) with a mean age of 23 years underwent tonsillectomy alone over a nine-month period, July 1998 to April 1999. The incidence of post-tonsillectomy haemorrhage in the synchronous nasal surgery group was 12.7 per cent (9 out of 71) compared with four per cent (16 out of 398) in the tonsillectomy alone group (p <0.01, Chi squared test). There was no difference in hospital stay between the two groups (mean two days). Synchronous nasal surgery increases the incidence of post tonsillectomy haemorrhage. PMID- 14561359 TI - Management of thyroglossal tract disease after failed Sistrunk's procedure. AB - Sistrunk's procedure for thyroglossal duct remnants has a very high success rate, there remains the occasional patient, however, that will have recurrent disease despite a competently performed operation. Applied anatomy and embryology proffer a solution to this problem. Extending the Sistrunk operation, with an anterior wide local excision remaining within normal tissue, enables removal of the entire thyroglossal tract remnant. A retrospective case note review was conducted to study our experience using this extended procedure to treat patients with thyroglossal tract disease that had recurred after a previous Sistrunk's operation. Six patients aged from five to 33 years were included in the study. There was one recurrence and the complication rate was comparable to the standard operation. It was concluded that a wide local excision is a valuable extension of the Sistrunk operation for the management of recurrent disease. PMID- 14561360 TI - Treatment of primary mucosal head and neck squamous cell carcinoma using photodynamic therapy: results after 25 treated cases. AB - The use of photodynamic therapy for the treatment of malignant and non-malignant conditions is increasing. This paper demonstrates the efficacy of a second generation photosensitizer, Foscan, in the primary treatment of a wide range of mucosal head and neck squamous cell carcinomas. Tumours ranged in stage from T(1) to T(3). A complete response to primary treatment was seen in 19/21 patients (90 per cent). In laryngeal cancer recurrent after radical radiotherapy, one out of four patients treated obtained a complete response (25 per cent). Six patients (24 per cent) required surgery after photodynamic therapy, for local recurrence or dysplasia. Mean follow up was for 27.3 months (standard deviation 20.6 months). PMID- 14561361 TI - Technique for secondary tracheo-oesophageal puncture in difficult necks. AB - A secondary tracheo-oesophageal puncture is required for voice restoration post laryngectomy either for reasons of closure of the primary fistula or as a planned secondary procedure. A method of doing this safely is described using an endotracheal tube and a flexible nasopharyngoscope in especially difficult cases. PMID- 14561362 TI - Medical treatment of nasal squamous papilloma with imiquimod cream. AB - Warts are common lesions. The prevalence and incidence of squamous papilloma of the nasal vestibulum are not known, and it is not certain how often human papilloma viruses are involved. Numerous medical or surgical treatment modalities have been reported in the literature with variable, sometimes unsatisfactory results. Imiquimod cream is a topically applied immunomodulator. It is used for warts and other skin tumours on different locations of the body. We report results of six patients treated with imiquimod five per cent cream for nasal squamous papilloma. PMID- 14561363 TI - New sinus culture applicator (Yariktas applicator). AB - Identification of the pathogen is crucial for the selection of an appropriate antibiotic in the management of sinusitis. We describe an original instrument that provides contamination-free culture from the sinus in endoscopic sinus surgery. PMID- 14561364 TI - Atypical keratosis obturans. AB - Keratosis obturans is a rare condition characterized by the accumulation of desquamated keratin material in the bony portion of the external auditory canal. Classically, it is reported to present with severe otalgia, conductive deafness and global widening of the canal. A case of keratosis obturans is described in which the principal symptom was a metallic taste and the main finding was extensive erosion of the hypotympanum with exposure of the facial nerve and the annulus of the tympanic membrane. This presenting symptom and resorption pattern are atypical of keratosis obturans and have not been documented previously. PMID- 14561365 TI - Supratentorial arachnoid cyst mimicking a Meniere's disease attack. AB - Arachnoid cysts (AC) often present with symptoms depending on their anatomical location within the skull; while supratentorial cysts grow causing relatively few symptoms, infratentorial ones may impair liquor circulation at the level of the fourth ventricle, giving rise to intracranial hypertension, or may stretch the complex nerve network in the cerebello-pontine angle. We report the singular clinical history of a 54-year-old male, who suddenly began to feel dizzy while sitting at his workplace, complaining of left tinnitus and aural fullness, in a classic clinical picture of Meniere's disease. The anomalous findings at otoneurological examination (markedly the left deviation at the Fukuda test) made a magnetic resonance image (MRI) scan mandatory and a huge AC was found in the left fronto-parietal lobe. The mass provoked an evident clockwise rotation of the brainstem that we suppose to be the cause of acoustico-facial bundle stretching explaining the vestibular symptomatology. PMID- 14561366 TI - Salivary duct carcinoma of submandibular gland with trigeminal nerve invasion to intracranium. AB - Salivary duct carcinoma is a rare and invasive malignant tumour with rapid distant metastasis and dismal prognosis. Clinically, perineural invasion of the salivary duct carcinoma is commonly noted. Here, we present a case of salivary duct carcinoma of submandibular gland origin with perineural invasion of the trigeminal nerve proximal to the intracranium, that was well demonstrated by a magnetic resonance image (MRI) and was consistent with the clinical presentation. This case received radical resection and radiotherapy with inclusion of the skull base within the field. There was no tumour recurrence and distant metastasis 24 months post-operatively. PMID- 14561367 TI - Intramuscular capillary hamartoma of the tongue. AB - Intramuscular vascular lesions in the tongue are rare and are usually the capillary form of haemangioma. A case is described of a lingual mass that resembled intramuscular haemangioma but had unique histological features that do not appear to have been described previously. The term intramuscular capillary hamartoma is suggested to describe it. PMID- 14561368 TI - Ewings' sarcoma of the mandible. AB - Ewings' sarcoma involving the facial bones is rare although it is the second most frequent bone malignancy. For the diagnosis a biopsy is needed. Currently the treatment is the combination of chemotherapy and radiotherapy with surgery indicated in only a few instances. We report a case of Ewing's sarcoma of the mandible and describe its clinicopathologic features and the treatment of this disease. PMID- 14561369 TI - Benign ectopic submandibular thyroid with a normotopic multinodular goitre. AB - Ectopic submandibular thyroid tissue is a rare entity and poses difficult diagnostic and management problems. The first case of ectopic submandibular thyroid with a normotopic multinodular goitre is presented PMID- 14561370 TI - Re: Citation analysis of otorhinolaryngology journals. PMID- 14561372 TI - The MS Forum Pan Asian Conference. PMID- 14561371 TI - Early management of multiple sclerosis: report from the MS Forum 15th Modern Management Workshop. PMID- 14561373 TI - Clinical management of cognitive impairment in multiple sclerosis: a review of current evidence. AB - Over the past decade, there has been considerable interest in assessing the importance and functional impact of cognitive impairment in multiple sclerosis (MS), its clinico-magnetic resonance imaging correlates, and possible approaches to clinical management. There is preliminary evidence that treatment with both disease-modifying and symptomatic agents may positively influence the cognitive outcome of the patient, but few studies have assessed the efficacy of cognitive rehabilitation programmes using an experimental design. To make further progress in the field of neuropsychological diagnosis, it is essential to validate sensitive, cost-effective and reliable screening instruments that can be used, in clinical settings, to identify patients appropriate for extensive neuropsychological assessment, and finalize tailor-made rehabilitation techniques for MS patients. PMID- 14561374 TI - Perspectives in gene therapy for MS. AB - The blood-brain barrier limits the therapeutic efficacy of systemic administration of anti-inflammatory and/or neuroprotective molecules to patients affected by immune-mediated inflammatory demyelinating diseases of the central nervous system (CNS) such as multiple sclerosis. Drug delivery to the CNS using non-replicative viral vectors may represent a valid alternative therapeutic strategy. Gene therapy for multiple sclerosis might include different "human grade" vectors, which could be used to deliver anti-inflammatory molecules as well as neuroprotective agents into the CNS in a flexible and useful way. These potential "therapeutic" vectors would have different life spans, tissue tropism and infectivity rates. PMID- 14561375 TI - Primary progressive MS. PMID- 14561376 TI - Report from the 55th Annual Meeting of the American Academy of Neurology (AAN). PMID- 14561377 TI - Oestrogen therapy for multiple sclerosis: not the way forward. PMID- 14561379 TI - Hormones and pregnancy. PMID- 14561380 TI - Human herpesvirus 6 and MS disease activity. PMID- 14561381 TI - The interaction of MS and pregnancy: a critical review. AB - Previous research on the effects of pregnancy on multiple sclerosis (MS) is somewhat flawed, and well-controlled, well-designed studies are needed to validate trial findings. In general, pregnancy appears to have a protective effect on MS course, with fewer, less severe relapses, especially in the third trimester. The exacerbation rate is increased in the first 3 months after delivery, but the overall relapse rate is no different to that observed in non pregnant MS patients. A woman's past history of relapses may be the best indicator of clinical course during and immediately after pregnancy. Pregnancy does not appear to affect the long-term course of MS. PMID- 14561382 TI - The immune system and hormones: review and relevance to pregnancy and contraception in women with MS. AB - Two-thirds of multiple sclerosis (MS) patients are women, and the average age of onset overlaps the childbearing years. Clinicians are frequently asked, therefore, about the most appropriate form of contraception and the risk of an MS relapse/exacerbation during pregnancy and the post-partum period. This paper reviews the literature on the immune system and the effects of pregnancy, oral contraceptives and hormone replacement therapy on MS. PMID- 14561383 TI - Reproductive counselling for MS: a rationale. AB - Many questions arise when counselling multiple sclerosis (MS) patients on the effects of MS on pregnancy, and vice versa. Reassurance can often be given regarding contraception, fertility, pregnancy management, pregnancy outcome, and the risk of the child developing MS. Much more information is needed, however, on the effects and implications of MS therapies on pregnancy and breast-feeding. PMID- 14561385 TI - Reproductive counselling in MS: a guide for healthcare professionals. PMID- 14561384 TI - Hormone-based therapies in MS. AB - It is well established in multiple sclerosis (MS) that the frequency of relapses decreases in late pregnancy, but increases in the post-partum period. This protective effect is probably due to hormonal and immune response changes induced by pregnancy. Oestrogens may be responsible, at least in part, for this protective effect. The potential use of oestrogens as therapy for MS is discussed. PMID- 14561386 TI - Distinguishing between immunophenotype and genotype in non-Hodgkin's lymphomas. PMID- 14561387 TI - Improved outcomes from dose-dense adjuvant chemotherapy for breast cancer with growth factor support. PMID- 14561388 TI - Hematopoietic growth factors in myelodysplastic syndromes. AB - The myelodysplastic syndromes (MDS) are a heterogeneous group of clonal hematopoietic stem cell disorders characterized clinically by refractory cytopenias in one or more myeloid cell lines and an increased probability of transformation to acute leukemia. Supportive care remains the mainstay of therapy in MDS and frequently includes monotherapy and combination therapy with hematopoietic growth factors, such as erythropoietin, granulocyte colony stimulating factor, and granulocyte macrophage colony-stimulating factor. Clinical trials have demonstrated the ability of growth factors to improve neutropenia and anemia in selected patients with MDS, which may have clinical, quality-of-life, and economic benefits for patients even though overall survival has not been improved. This paper reviews the role of hematopoietic growth factors in the treatment of MDS. PMID- 14561389 TI - Erythropoietin and renal failure. AB - Erythropoietic therapy has transformed the management of the anemia associated with chronic renal failure over the past decade. The first agents that proved effective in stimulating erythropoiesis were the epoetins (alfa and beta), which are recombinant human erythropoietins (EPOs). The EPO molecule was recently modified using site-directed mutagenesis to produce darbepoetin alfa with a longer circulating half-life in vivo. The development of epoetin-induced pure red cell aplasia associated with anti-EPO antibodies shook the nephrology community last year, and gradually new information is emerging on the cause of this rare but important condition. Pure red cell aplasia has provoked new discussions on the best route (intravenously or subcutaneously) for administering erythropoietic therapy. Further research has been published regarding some of the more traditional, although still hotly debated, issues concerning renal anemia management, such as target hemoglobin, factors affecting the response to EPO, and adjuvant therapy to EPO. This review addresses these topics and focuses largely on the publications relevant to these areas of clinical anemia management. PMID- 14561390 TI - Erythropoietin is a multifunctional tissue-protective cytokine. AB - Erythropoietin (EPO) was originally identified as a hormone produced by the adult kidney to facilitate optimum delivery of oxygen to tissue beds by adjustment of the circulating erythrocyte mass. The cloning of the EPO gene, subsequent production of recombinant protein, and successful introduction into clinical practice for the treatment of the anemia of renal failure is a triumph of biotechnology. However, molecular biologic studies have established that EPO is a member of the cytokine superfamily, with significant homology to mediators of growth and inflammation. Therefore, it is not surprising that additional biologic functions for this protein have been identified. Results from studies have shown that EPO and its receptor are widely expressed in embryonic and adult tissues, including the central nervous system, gut, kidney, muscle (eg, smooth, skeletal, and heart), uterus, retina, pancreas, gonads, and lung. We review the evidence that EPO and its receptor function as a paracrine/autocrine system to mediate the protection of tissues subjected to metabolic stress. PMID- 14561391 TI - Cost effectiveness of myeloid growth factors in cancer chemotherapy. AB - Healthcare costs continue to rise and hospitalization represents the single largest component of direct medical costs associated with cancer care. Neutropenia and its complications, including febrile neutropenia (FN), remain the major dose-limiting toxicity of systemic cancer chemotherapy. Although under reported, FN often occurs early in the course of chemotherapy and contributes substantially to the morbidity, mortality, and cost of treatment. The risk of FN and its complications are associated with treatment intensity, age, and various comorbidities. Myeloid growth factors (MGFs) have been used effectively in a variety of clinical settings to prevent or treat FN and assist patients receiving dose-intensive chemotherapy with or without stem cell support. A meta-analysis of the available randomized controlled trials has confirmed the efficacy of prophylactic MGFs. The cost of these agents, along with their large-scale clinical use, has prompted several economic investigations. Economic models based on measures of resource use derived from randomized controlled trials have provided estimates of expected treatment costs, along with FN risk threshold estimates for the cost-saving use of prophylactic MGF. Recent studies have demonstrated the potential value of targeting MGFs toward patients at greatest risk based on accurate and valid predictive models. Although an emerging role has become apparent for MGFs in managing adult leukemia and supporting high-dose therapy with stem cell transplantation in adults, their value in the support of children in these settings remains unclear. Continuing clinical and economic evaluation, along with an updating of clinical practice guidelines because of rapid technologic and clinical advances, is encouraged. PMID- 14561392 TI - Prophylactic use of myelopoietic growth factors in children after myelosuppressive and myeloablative therapy. AB - Judicious use of myelopoietic colony-stimulating factors is not well defined and is the source of ongoing controversy in the pediatric patient population. Prophylactic colony-stimulating factors may provide little clinically relevant benefits in children with hematologic malignancies or solid tumors after myelosuppressive chemotherapy. Although several studies demonstrated that recombinant human granulocyte colony-stimulating factor and granulocyte macrophage colony-stimulating factor accelerate neutrophil engraftment after myeloablative therapy and stem cell transplantation, their use did not alter morbidity or mortality after stem cell transplantation and has been associated with delayed platelet engraftment. Large prospective randomized multicenter studies are necessary to demonstrate an improvement in infectious complications, duration of hospitalization, overall survival, and cost-benefit ratio with the prophylactic use of myelopoietic growth factors after myelosuppressive or myeloablative chemotherapy in children and adolescents. Additional pediatric studies comparing the safety, efficacy, and cost effectiveness of granulocyte colony-stimulating factor versus granulocyte macrophage colony-stimulating factor are needed. PMID- 14561393 TI - The Canadian Universal Leukoreduction Program. PMID- 14561394 TI - Pathogen reduction technology: methods, status of clinical trials, and future prospects. AB - Transfusion-transmitted infections caused by viruses, bacteria, and protozoa have been extensively reported for the past three decades. In the early 1980s, the HIV epidemic heightened the focus on transfusion safety and demonstrated the potential for new pathogens to enter the donor population and rapidly spread through the transfusion of blood components. The foundation for the prevention of transfusion-transmitted infections has been donor screening and testing, and significant advances have been made to reduce the risk of transfusion-associated infection. However, despite these efforts, transfusion-transmitted infections continue to be reported, and new infectious agents, such as the West Nile virus, continue to enter the donor population with transmission to recipients. For the past decade, several technologies to inactivate infectious pathogens that may contaminate donor blood have been developed and several of these methods have been introduced into clinical practice. These technologies offer the potential for a paradigm shift to further improve the safety of blood transfusion. PMID- 14561395 TI - Alternative oxygen therapeutics: products, status of clinical trials, and future prospects. AB - Alternative oxygen therapeutics are oxygen carriers that can potentially replace red blood cell transfusion in clinical instances in which banked blood is unavailable or unsafe. Most of the candidate solutions have been based on hemoglobin, derived from animals, outdated banked human blood, or recombinant systems. Other solutions are based on perfluorocarbons. In contrast to hemoglobin, which chemically binds oxygen, perfluorocarbons carry oxygen as a dissolved gas. Most of the products of either category developed over the past two decades have failed attempts at regulatory approval for clinical use, but their development has led to a large body of research that has defined the way oxygen is normally delivered to tissues. From this research, a new generation of products is emerging, designed specifically to target oxygen to the tissues in most need. It is possible that one or more of the earlier products may be approved by regulatory agencies in 1 or 2 years. However, a universally compatible and safe product most likely will be unavailable for several more years. PMID- 14561396 TI - Vector-borne illnesses and the safety of the blood supply. AB - The protection of patients from diseases carried in blood transfusions is an ongoing effort. The viruses that cause long-term human infection and death have received much of the attention in the United States and testing has significantly diminished the risk of infection from a transfusion. As the risk of these diseases has decreased, other transfusion-transmitted organisms with a lower incidence in the community or newer diseases with rapidly expanding endemic areas are receiving additional attention. One group of these infections are infections in which the normal route of human infection is a vector. PMID- 14561397 TI - Transfusion errors: scope of the problem, consequences, and solutions. AB - For more than 25 years, ABO transfusion errors in the United States have caused more noninfectious transfusion deaths each year than any other cause. The US ABO error rate is between 1:12,000 and 1:19,000, with a fatality rate between 1:800,000 and 1:1.3 million. In the past 15 years, international research has increased to determine the frequency and nature of the slips and mistakes that result in transfusion errors and possible solutions to reduce them. Solutions include changes in human processes, introducing technologic improvements, and involving professional societies to create national or international performance standards to serve as benchmarks for comparison. PMID- 14561398 TI - Granulocyte transfusion: revisited. AB - Neutrophils are the immune system's main cellular defense against bacterial and fungal infections. Transfusion of granulocytes has been considered a therapeutic modality for severe bacterial and fungal infections in patients with prolonged neutropenia and with functional neutrophil disorders. Good theoretic and experimental evidence demonstrating granulocyte transfusion efficacy exists in preventing and treating severe infection. However, clinical evidence has been more difficult to interpret, with efficacy equivocal in many studies and further trials hindered by limitations in collecting adequate doses of leukocytes from healthy steroid-mobilized donors. The development and use of granulocyte colony stimulating factor to stimulate normal donors has generated renewed interest in granulocyte transfusions. In clinical studies, granulocyte colony-stimulating factor has markedly enhanced the yield of leukocytes collected from normal donors, which may improve clinical outcomes in patients with severe infections and neutropenia who receive granulocyte transfusions. Preliminary clinical evidence, when correct granulocyte dose per patient body weight is optimized, suggests efficacy. However, well-designed randomized clinical trials are necessary to definitively establish granulocyte transfusions as a viable therapeutic modality in the treatment of severe bacterial and fungal infections in patients with functional neutrophil disorders or neutropenia. PMID- 14561399 TI - Centromere silencing and function in fission yeast is governed by the amino terminus of histone H3. AB - BACKGROUND: Centromeric domains often consist of repetitive elements that are assembled in specialized chromatin, characterized by hypoacetylation of histones H3 and H4 and methylation of lysine 9 of histone H3 (K9-MeH3). Perturbation of this underacetylated state by transient treatment with histone deacetylase inhibitors leads to defective centromere function, correlating with delocalization of the heterochromatin protein Swi6/HP1. Likewise, deletion of the K9-MeH3 methyltransferase Clr4/Suvar39 causes defective chromosome segregation. Here, we create fission yeast strains retaining one histone H3 and H4 gene; the creation of these strains allows mutation of specific N-terminal tail residues and their role in centromeric silencing and chromosome stability to be investigated. RESULTS: Reduction of H3/H4 gene dosage to one-third does not affect cell viability or heterochromatin formation. Mutation of lysines 9 or 14 or serine 10 within the amino terminus of histone H3 impairs centromere function, leading to defective chromosome segregation and Swi6 delocalization. Surprisingly, silent centromeric chromatin does not require the conserved lysine 8 and 16 residues of histone H4. CONCLUSIONS: To date, mutation of conserved N terminal residues in endogenous histone genes has only been performed in budding yeast, which lacks the Clr4/Suvar39 histone methyltransferase and Swi6/HP1. We demonstrate the importance of conserved residues within the histone H3 N terminus for the maintenance of centromeric heterochromatin in fission yeast. In sharp contrast, mutation of two conserved lysines within the histone H4 tail has no impact on the integrity of centromeric heterochromatin. Our data highlight the striking divergence between the histone tail requirements for the fission yeast and budding yeast silencing pathways. PMID- 14561400 TI - A self-sustaining, light-entrainable circadian oscillator in the Drosophila brain. AB - BACKGROUND: The circadian clock of Drosophila is able to drive behavioral rhythms for many weeks in continuous darkness (DD). The endogenous rhythm generator is thought to be generated by interlocked molecular feedback loops involving circadian transcriptional and posttranscriptional regulation of several clock genes, including period. However, all attempts to demonstrate sustained rhythms of clock gene expression in DD have failed, making it difficult to link the molecular clock models with the circadian behavioral rhythms. Here we restricted expression of a novel period-luciferase transgene to certain clock neurons in the Drosophila brain, permitting us to monitor reporter gene activity in these cells in real-time. RESULTS: We show that only a subset of the previously described pacemaker neurons is able to sustain PERIOD protein oscillations after 5 days in constant darkness. In addition, we identified a sustained and autonomous molecular oscillator in a group of clock neurons in the dorsal brain with heretofore unknown function. We found that these "dorsal neurons" (DNs) can synchronize behavioral rhythms and that light input into these cells involves the blue-light photoreceptor cryptochrome. CONCLUSIONS: Our results suggest that the DNs play a prominent role in controlling locomotor behavior when flies are exposed to natural light-dark cycles. Analysis of similar "stable mosaic" transgenes should help to reveal the function of the other clock neuronal clusters within the fly brain. PMID- 14561401 TI - Radial patterning of Arabidopsis shoots by class III HD-ZIP and KANADI genes. AB - BACKGROUND: Shoots of all land plants have a radial pattern that can be considered to have an adaxial (central)-abaxial (peripheral) polarity. In Arabidopsis, gain-of-function alleles of PHAVOLUTA and PHABULOSA, members of the class III HD-ZIP gene family, result in adaxialization of lateral organs. Conversely, loss-of-function alleles of the KANADI genes cause an adaxialization of lateral organs. Thus, the class III HD-ZIP and KANADI genes comprise a genetic system that patterns abaxial-adaxial polarity in lateral organs produced from the apical meristem. RESULTS: We show that gain-of-function alleles of REVOLUTA, another member of the class III HD-ZIP gene family, are characterized by adaxialized lateral organs and alterations in the radial patterning of vascular bundles in the stem. The gain-of-function phenotype can be obtained by changing only the REVOLUTA mRNA sequence and without changing the protein sequence; this finding indicates that this phenotype is likely mediated through an interference with microRNA binding. Loss of KANADI activity results in similar alterations in vascular patterning as compared to REVOLUTA gain-of-function alleles. Simultaneous loss-of-function of PHABULOSA, PHAVOLUTA, and REVOLUTA abaxializes cotyledons, abolishes the formation of the primary apical meristem, and in severe cases, eliminates bilateral symmetry; these phenotypes implicate these three genes in radial patterning of both embryonic and postembryonic growth. CONCLUSIONS: Based on complementary vascular and leaf phenotypes of class III HD ZIP and KANADI mutants, we propose that a common genetic program dependent upon miRNAs governs adaxial-abaxial patterning of leaves and radial patterning of stems in the angiosperm shoot. This finding implies that a common patterning mechanism is shared between apical and vascular meristems. PMID- 14561402 TI - Cdk2 knockout mice are viable. AB - BACKGROUND: Cyclin-dependent kinases (Cdks) and their cyclin regulatory subunits control cell growth and division. Cdk2/cyclin E complexes are thought to be required because they phosphorylate the retinoblastoma protein and drive cells through the G1/S transition into the S phase of the cell cycle. In addition, Cdk2 associates with cyclin A, which itself is essential for cell proliferation during early embryonic development. RESULTS: In order to study the functions of Cdk2 in vivo, we generated Cdk2 knockout mice. Surprisingly, these mice are viable, and therefore Cdk2 is not an essential gene in the mouse. However, Cdk2 is required for germ cell development; both male and female Cdk2(-/-) mice are sterile. Immunoprecipitates of cyclin E1 complexes from Cdk2(-/-) spleen extracts displayed no activity toward histone H1. Cyclin A2 complexes were active in primary mouse embryonic fibroblasts (MEFs), embryo extracts and in spleen extracts from young animals. In contrast, there was little cyclin A2 kinase activity in immortalized MEFs and spleen extracts from adult animals. Cdk2(-/-) MEFs proliferate but enter delayed into S phase. Ectopic expression of Cdk2 in Cdk2(-/-) MEFs rescued the delayed entry into S phase. CONCLUSIONS: Although Cdk2 is not an essential gene in the mouse, it is required for germ cell development and meiosis. Loss of Cdk2 affects the timing of S phase, suggesting that Cdk2 is involved in regulating progression through the mitotic cell cycle. PMID- 14561404 TI - Auditory and action semantic features activate sensory-specific perceptual brain regions. AB - Traditionally, concepts were considered propositional, amodal, and verbal in nature (for review, see ). Recent findings, however, suggest that conceptual knowledge is divisible into different types (L. Wu and L.W. Barsalou, personal communication, ) and that each type may be linked to specific sensory and motor processes. This implies that sensory processing regions of the brain may also process concepts. In fact, there is some neuroimaging evidence that conceptual information does activate perceptual brain regions and that there is a correspondence between knowledge type and the region being activated. In the following experiment, using a training technique developed in previous studies, participants verbally learned associations between novel objects and conceptual features. The objective was to create objects that were associated with features from only one knowledge type, something that does not occur with common objects. During a visual task that did not require retrieval of learned associations, the superior temporal gyrus, which responds well to sounds, was preferentially activated by objects associated with auditory features (e.g., buzzes). Likewise, the posterior superior temporal sulcus, which responds well to motion, was preferentially activated by objects associated with "action" features (e.g., hops). These findings support the theory that knowledge is grounded in perception. PMID- 14561403 TI - Dpp signaling silences bam transcription directly to establish asymmetric divisions of germline stem cells. AB - Stem cells execute self-renewing and asymmetric cell divisions in close association with stromal cells that form a niche. The mechanisms that link stromal cell signaling to self-renewal and asymmetry are only beginning to be identified, but Drosophila oogenic germline stem cells (GSCs) have emerged as an important model for studying stem cell niches. A member of the Bone Morphogenetic Protein (BMP) ligand family, Decapentaplegic (Dpp), sustains ovarian GSCs by suppressing differentiation in the stem cell niche (Figure 1A). Dpp overexpression expands the niche, blocks germ cell differentiation, and causes GSC hyperplasty. Here, we show that the bag-of-marbles (bam) differentiation factor is the principal target of Dpp signaling in GSCs; ectopic bam expression restores differentiation even when Dpp is overexpressed. We show that the transcriptional silencer element in the bam gene integrates Dpp control of bam expression. Finally and most significantly, we demonstrate for the first time that Dpp signaling regulates bam expression directly since the bam silencer element is a strong binding site for the Drosophila Smads, Mad and Medea. These studies provide a simple mechanistic explanation for how stromal cell signals regulate both the self-renewal and asymmetric fates of the products of stem cell division. PMID- 14561405 TI - The meiosis I-to-meiosis II transition in mouse oocytes requires separase activity. AB - Faithful segregation of homologous chromosomes during the first meiotic division is essential for further embryo development. The question at issue is whether the same mechanisms ensuring correct separation of sister chromatids in mitosis are at work during the first meiotic division. In mitosis, sister chromatids are linked by a cohesin complex holding them together until their disjunction at anaphase. Their disjunction is mediated by Separase, which cleaves the cohesin. The activation of Separase requires prior degradation of its associated inhibitor, called securin. Securin is a target of the APC/C (Anaphase Promoting Complex/Cyclosome), a cell cycle-regulated ubiquitin ligase that ubiquitinates securin at the metaphase-to-anaphase transition and thereby targets it for degradation by the 26S proteasome. After securin degradation, Separase cleaves the cohesins and triggers chromatid separation, a prerequisite for anaphase. In yeast and worms, the segregation of homologous chromosomes in meiosis I depends on the APC/C and Separase activity. Yet, it is unclear if Separase is required for the first meiotic division in vertebrates because APC/C activity is thought to be dispensable in frog oocytes. We therefore investigated if Separase activity is required for correct chromosome segregation in meiosis I in mouse oocytes. PMID- 14561406 TI - Asymmetries in stimulus comparisons by monkey and man. AB - Similarity is a core concept in theories of object recognition, categorization, and reasoning. It is often conceptualized as a geometric distance in a multidimensional stimulus space. However, research in humans has revealed that similarity judgments involve more than a simple distance calculation and tend to be asymmetric when stimuli differ in factors such as prototypicality. For example, most people judge 99 to be more similar to 100 than 100 to 99. Up to now, it was not known whether such asymmetries might also occur in nonhuman subjects. This study reveals asymmetries in the pattern of errors made by four rhesus monkeys in a temporal same/different task. Monkeys usually perceived a smaller difference between two different stimuli when the first stimulus in a trial was less prototypical than the second, just as what was found previously for human subjects. The pattern of asymmetries differed between monkeys, and a control study showed that such variability is also present for human subjects. We propose that known neurophysiological mechanisms can account for asymmetry in the stimulus comparisons of both species. Thus, seemingly complex phenomena that occur when human subjects rate stimulus similarity are also present in macaques' similarity judgments and could be based on relatively simple mechanisms. PMID- 14561407 TI - Genetic control of methyl halide production in Arabidopsis. AB - Methyl chloride (CH(3)Cl) and methyl bromide (CH(3)Br) are the primary carriers of natural chlorine and bromine, respectively, to the stratosphere, where they catalyze the destruction of ozone, whereas methyl iodide (CH(3)I) influences aerosol formation and ozone loss in the boundary layer. CH(3)Br is also an agricultural pesticide whose use is regulated by international agreement. Despite the economic and environmental importance of these methyl halides, their natural sources and biological production mechanisms are poorly understood. Besides CH(3)Br fumigation, important sources include oceans, biomass burning, tropical plants, salt marshes, and certain crops and fungi. Here, we demonstrate that the model plant Arabidopsis thaliana produces and emits methyl halides and that the enzyme primarily responsible for the production is encoded by the HARMLESS TO OZONE LAYER (HOL) gene. The encoded protein belongs to a group of methyltransferases capable of catalyzing the S-adenosyl-L-methionine (SAM) dependent methylation of chloride (Cl(-)), bromide (Br(-)), and iodide (I(-)) to produce methyl halides. In mutant plants with the HOL gene disrupted, methyl halide production is largely eliminated. A phylogenetic analysis with the HOL gene suggests that the ability to produce methyl halides is widespread among vascular plants. This approach provides a genetic basis for understanding and predicting patterns of methyl halide production by plants. PMID- 14561408 TI - A coat of filamentous actin prevents clustering of late-endosomal vacuoles in vivo. AB - The endocytic pathway depends on the actin cytoskeleton. Actin contributes to internalization at the plasma membrane and to subsequent trafficking steps like propulsion through the cytoplasm, fusion of phagosomes with early endosomes, and transport from early to late endosomes. In vitro studies with mammalian endosomes and yeast vacuoles implicate actin in membrane fusion. Here, we investigate the function of the actin coat that surrounds late endosomes in Dictyostelium. Latrunculin treatment leads to aggregation of these endosomes into grape-like clusters and completely blocks progression of endocytic marker. In addition, the cells round up and stop moving. Because this drug treatment perturbs all actin assemblies in the cell simultaneously, we used a novel targeting approach to specifically study the function of the cytoskeleton in one subcellular location. To this end, we constructed a hybrid protein targeting cofilin, an actin depolymerizing protein, to late endosomes. As a consequence, the endosomal compartments lost their actin coats and aggregated, but these cells remained morphologically normal, and the kinetics of endocytic marker trafficking were unaltered. Therefore, the actin coat prevents the clustering of endosomes, which could be one safeguard mechanism precluding their docking and fusion. PMID- 14561409 TI - Formin leaky cap allows elongation in the presence of tight capping proteins. AB - Formins, characterized by formin homology domains FH1 and FH2, are required to assemble certain F-actin structures including actin cables, stress fibers, and the contractile ring. FH1FH2 in a recombinant fragment from a yeast formin (Bni1p) nucleates actin filaments in vitro. It also binds to the filament barbed end where it appears to act as a "leaky" capper, slowing both polymerization and depolymerization by approximately 50%. We now find that FH1FH2 competes with tight capping proteins (including gelsolin and heterodimeric capping protein) for the barbed end. We also find that FH1FH2 forms a tetramer. The observation that this formin protects an end from capping but still allows elongation confirms that it is a leaky capper. This is significant because a nucleator that protects a new barbed end from tight cappers will increase the duration of elongation and thus the total amount of F-actin. The ability of FH1FH2 to dimerize probably allows the formin to walk processively with the barbed end as the filament elongates. PMID- 14561410 TI - Effects of low-spatial frequency components of fearful faces on fusiform cortex activity. AB - Emotive faces elicit neural responses even when they are not consciously perceived. We used faces hybridized from spatial frequency-filtered individual stimuli to study processing of facial emotion. Employing event-related functional magnetic resonance imaging (fMRI), we show enhanced fusiform cortex responses to hybrid faces containing fearful expressions when such emotional cues are present in the low-spatial frequency (LSF) range. Critically, this effect is independent of whether subjects use LSF or high-spatial frequency (HSF) information to make gender judgments on the hybridized faces. The magnitude of this fusiform enhancement predicts behavioral slowing in response times when participants report HSF information of the hybrid stimulus in the presence of fear in the unreported LSF components. Thus, emotional modulation of a face-responsive region of fusiform is driven by the low-frequency components of the stimulus, an effect independent of subjects' reported perception but evident in an incidental measure of behavioral performance. PMID- 14561412 TI - Top scientists back human cloning ban. PMID- 14561411 TI - Fgf signaling controls the number of phalanges and tip formation in developing digits. AB - Tetrapods have two pairs of limbs, each typically with five digits, each of which has a defined number of phalanges derived from an archetypal formula. Much progress has been made in understanding vertebrate limb initiation and the patterning processes that determine digit number in developing limb buds, but little is known about how phalange number is controlled. We and others previously showed that an additional phalange can be induced in a chick toe if sonic hedgehog protein is applied in between developing digit primordia. Here we show that formation of an additional phalange is associated with prolonged Fgf8 expression in the overlying apical ridge and that an Fgf Receptor inhibitor blocks its formation. The additional phalange is produced by elongation and segmentation of the penultimate phalange, suggesting that the digit tip forms when Fgf signaling ceases by a special mechanism, possibly involving Wnt signaling. Consistent with this, Fgfs inhibit tip formation whereas attenuation of Fgf signaling induces tip formation prematurely. We propose that duration of Fgf signaling from the ridge, responsible for elongation of digit primordia, coupled with a characteristic periodicity of joint formation, generates the appropriate number of phalanges in each digit. We also propose that the process that generates the digit tips is independent of that which generates more proximal phalanges. This has implications for understanding human limb congenital malformations and evolution of digit diversity. PMID- 14561413 TI - Fungal familiarities. PMID- 14561414 TI - Q & A. AB - Gregory A. Petsko is Gyula and Katica Tauber Professor of Biochemistry and Chemistry and Director of the Rosenstiel Basic Medical Sciences Research Center at Brandeis University. He did his undergraduate work at Princeton and his graduate work as a Rhodes Scholar at Oxford University. He held faculty positions at Wayne State University School of Medicine and MIT before moving to Brandeis in 1990. A structural biologist, he is best known for his work, together with his colleague Dagmar Ringe, on the structural basis of enzyme catalytic power and the role of protein dynamics in protein function. He writes a regular opinion column for the journal Genome Biology. PMID- 14561415 TI - Speaking volumes. AB - Many have pondered the potential impact of Mendel's work if Darwin had come across it in his lifetime. Darwin found difficulty with a blending inheritance and natural selection but a new letter suggests he was grappling with a more modern view. Nigel Williams reports. PMID- 14561416 TI - Male pregnancy. PMID- 14561417 TI - Fungal mating-type loci. PMID- 14561418 TI - Cell motility: deaf Drosophila keep the beat. AB - Intraflagellar transport is involved in the assembly of cilia and eukaryotic flagella. Two recent studies have shown that defects in intraflagellar transport prevent assembly of sensory cilia in Drosophila, leaving the fly deaf and uncoordinated. Surprisingly, the mutant sperm flagella have normal structure and motility. PMID- 14561419 TI - Mg2+ homeostasis: the Mg2+nificent TRPM chanzymes. AB - TRPM6 and TRPM7 are distinct from all other ion channels in that they are composed of linked channel and protein kinase domains. Recent studies demonstrate that these 'chanzymes' are essential for Mg(2+) homeostasis, which is critical for human health and cell viability. PMID- 14561420 TI - Motor physiology: a brain of two halves. AB - Brain changes after stroke suggest that undamaged areas may 'take over' the function of damaged regions. Recent studies using magnetic stimulation to disrupt the healthy human brain shed new light on the potential for dynamic compensation across the motor system. PMID- 14561421 TI - Cell turnover: flexible coupling meets the needs of development. AB - The identification of Drosophila genes that inhibit proliferation while simultaneously promoting apoptosis--decreasing cell number--or that promote proliferation while simultaneously inhibiting apoptosis--increasing cell number- has revealed new ways that cell birth and death may be coupled to meet the needs of development. PMID- 14561422 TI - Left-right asymmetry: nodal cilia make and catch a wave. AB - Asymmetric fluid flow in the mouse node initiates the development of left-right asymmetry. This flow is generated by motile cilia and is detected by immotile mechanosensory cilia, activating an asymmetric calcium spike. PMID- 14561423 TI - Microtubule assembly: catastrophe factors to the rescue. AB - Two new studies have shown how regulation of microtubule dynamics by members of the kinesin superfamily may guard against errors in spindle assembly and chromosome segregation. PMID- 14561424 TI - EGF receptor signaling: putting a new spin on eye development. AB - The coordinated polarization of cells within an epithelium is required for the development and function of some tissues. Recent work has shown that the EGF receptor signaling pathway plays a key role in establishing epithelial polarity in the compound eye of Drosophila. PMID- 14561425 TI - Lipid signalling: picking out the PIPs. AB - Many physiological targets have been suggested for polyphosphoinositol lipids, but two out of the three monophosphorylated PIPs appeared to be no more than metabolic precursors. Recent work has shown that they also have distinct binding proteins and functions. PMID- 14561426 TI - Brain processing of visual sexual stimuli in men with hypoactive sexual desire disorder. AB - Although hypoactive sexual desire disorder (HSDD) is a common condition and has long been hypothesized to result from malfunctions of the cerebral control mechanisms that adjust the level of sexual motivation, very little is known about the pathophysiology of this disorder. The primary objective was to identify in patients with HSDD brain regions where functional perturbations disrupt the regulation of sexual motivation. We used positron emission tomography to compare seven male patients with HSDD with eight healthy men on their regional cerebral blood flow responses to visual sexual stimuli (VSS) of graded intensity. Statistical Parametric Mapping was used to locate brain regions that demonstrated a differential activation (or deactivation) across the groups. Whereas in control subjects the medial orbitofrontal cortex showed a deactivation in response to VSS, in HSDD patients there was an abnormally maintained activity of this region, which has been implicated in the inhibitory control of motivated behavior. By contrast, the reverse pattern-activation in control subjects, deactivation or unchanged activity in patients-was found in the secondary somatosensory cortex and inferior parietal lobules, regions mediating emotional and motor imagery processes, as well as in those areas of the anterior cingulate gyrus and of the frontal lobes that are involved in premotor processes. PMID- 14561427 TI - Brain reactivity to specific symptom provocation indicates prospective therapeutic outcome in OCD. AB - A pertinent question in biological psychiatry is what differentiates responders and non-responders to pharmacological treatment. One possibility is that individual differences in the symptomatic spectrum as well as in the underlying biology of the disorder lead to the known 40% failure in pharmacological treatment. Our study aimed to maximize individual brain markers of obsessive compulsive disorder (OCD) by applying single photon emission computed tomography (SPECT) during a provoked symptomatic state prior to and following treatment. Four brain SPECT scans were obtained from 26 OCD patients prior to and at 6 months of sertraline treatment. At each time point, two SPECT scans were performed in a counterbalanced order of two specific states; one a symptom provoking condition and the other a relaxed condition. At 6 months of treatment, patients were divided into responders and non-responders according to a predetermined clinical criterion. Prospective responders showed significantly lower brain perfusion in the dorsal-caudal anterior cingulum and higher brain perfusion in the right caudate, when compared to non-responders, only during symptom provocation. When pre- and post-treatment scans during symptom provocation were compared, only responders showed significant change in brain response: increased perfusion in the left anterior temporal cortex and prefrontal cortex at 6 months' treatment. These findings suggest that obtaining functional brain imaging during specific symptom provocation emphasizes individual differences in brain reactivity. Thus can indicate prospective responders to symptom-related treatment in OCD and mark the relevant brain regions for effective response to treatment. PMID- 14561428 TI - Persistence of disturbed thalamic glucose metabolism in a case of Wernicke Korsakoff syndrome. AB - We report the case of a 40-year-old alcoholic male patient, hospitalized with an acute ataxia of stance and gait, ocular muscle weakness with nystagmus and a global apathetic-confusional state. After admission, an amnestic syndrome with confabulation was also observed and diagnosis of Wernicke-Korsakoff syndrome was made. Under treatment with intravenous thiamine, the patient recovered completely from gaze weakness and ataxia, whereas a severe amnestic syndrome persisted. Fluorodeoxyglucose (FDG) positron emission tomography (PET) showed bilateral thalamic and severe bilateral temporal-parietal hypometabolism resembling a pattern typical for Alzheimer's disease. Longitudinal assessment of the alcohol abstinent and thiamine-substituted patient revealed improvements of clinical state and neuropsychological performance that were paralleled by recovered cerebral glucose metabolism. In contrast to metabolic rates that increased between 7.1% (anterior cingulate, left) and 23.5% (parietal, left) in cortical areas during a 9-month remission period, thalamic glucose metabolism remained severely disturbed over time (change: left +0.2%, right +0.3%). PMID- 14561429 TI - Effects of behavior therapy on regional cerebral blood flow in obsessive compulsive disorder. AB - Very few functional neuroimaging studies have been performed on patients with obsessive-compulsive disorder (OCD) undergoing behavior therapy, even though it is recognized to be an effective treatment for this disorder. We measured the regional cerebral blood flow (rCBF) using the Xenon inhalation method in 31 treatment-refractory patients with OCD and the same number of age-matched normal controls. We also studied changes in rCBF in 22 OCD patients who had demonstrated a significant improvement after the behavior therapy. The OCD patients showed a significant bilateral elevation in the rCBF in the basal ganglia compared with the normal controls. After successful treatment, a significant decrease was found in the rCBF in the right head of the caudate nucleus that tended to correlate with clinical improvement. PMID- 14561430 TI - State-related thalamic changes during antipsychotic treatment in schizophrenia: preliminary observations. AB - Thalamic volumes and psychotic symptoms were assessed during psychotic exacerbation and during antipsychotic drug treatment. Reduction of psychotic symptoms (SAPS) during four weeks of treatment was highly correlated with volumetric expansion as measured by magnetic resonance imaging in both left and right thalamus [r(s)=0.75 and r(s)=0.82, respectively (both P<0.04)]. PMID- 14561431 TI - Hippocampal glucose metabolism is associated with cerebrospinal fluid estrogen levels in postmenopausal women with Alzheimer's disease. AB - Animal studies indicate that estrogens, such as 17beta-estradiol (E(2)), may enhance hippocampal metabolism and function. In postmenopausal Alzheimer's disease (AD) patients, cerebrospinal fluid (CSF) E(2) levels were significantly lower than in non-demented controls. This finding was inversely correlated with CSF beta-amyloid levels. To address the potential impact of this finding, E(2) levels in CSF were correlated with regional cerebral [18F]2-fluoro-2-deoxy-D glucose (FDG) uptake as measured using positron emission tomography (PET) in six postmenopausal AD patients. CSF E(2) levels were determined using an electro chemiluminescence-immunoassay on the Roche Elecsys 2010 immunoassay analyzer. Basic image processing was done by MEDx, using SPM routines for spatial normalization and statistics. CSF E(2) levels were significantly correlated with cerebral glucose metabolism in the left hippocampus. This is the first clinical study indicating an association between CSF E(2) concentration and hippocampal glucose metabolism in postmenopausal women with AD. PMID- 14561432 TI - Increased levels of serum basic fibroblast growth factor in schizophrenia. AB - Basic fibroblast growth factor (bFGF) is a multifunctional growth factor that has been implicated in a variety of neurodevelopmental processes. The aim of the present study was to examine whether bFGF contributes to the pathophysiology of schizophrenia. Serum bFGF levels in 40 patients with schizophrenia (15 drug-naive and 25 medicated patients) and in 40 age- and sex-matched healthy normal controls were measured. Serum bFGF levels were significantly higher in the medicated patients than in the normal controls. Analysis of partial correlation coefficients showed that the increased bFGF levels might not be attributable to antipsychotic medication. Although there was no significant overall difference in bFGF levels between drug-naive patients and normal controls, the bFGF levels in these patients significantly correlated with the severity of negative symptoms. Furthermore, we found a significant negative correlation between serum bFGF levels and the age of onset in the entire patient group. Our finding of elevated bFGF levels in the serum of patients with schizophrenia, especially in earlier age-of-onset cases considered to have more neurodevelopmental insults, suggests that bFGF abnormalities may be involved in the pathophysiology of schizophrenia. PMID- 14561433 TI - Implications of compulsive and impulsive traits for serotonin status in women with bulimia nervosa. AB - Studies of bulimia nervosa (BN) often report decreased brain serotonin (5 hydroxytryptamine: 5-HT) activity. Across populations, impulsivity has been linked to reduced 5-HT activity, but compulsivity has been associated (at least inconsistently) with an increase. We therefore became interested in the association between behavioral-trait variations and 5-HT status in BN. In 56 bulimic and 29 non-bulimic women, we measured eating symptoms, personality traits, platelet paroxetine binding, and neuroendocrine responses following oral meta-chlorophenylpiperazine (m-CPP). Relative to normal eaters, bulimic women showed reduced density (Bmax) of platelet paroxetine-binding sites, blunted prolactin (PRL) responses following m-CPP, and (as a trend) lower basal PRL levels. However, after effects of binge-purge frequencies, body mass, and other extraneous factors were controlled, PRL levels at baseline and other moments in the serial sampling varied systematically with presence of impulsive and compulsive traits. PRL was generally low in BN, but 'high-compulsive'/'low impulsive' traits were associated with higher (normal-range) PRL values. Comparable trait-related variations were not observed on paroxetine-binding indices. Our findings suggest that 5-HT status in BN may correspond to impulsive or compulsive traits, and they encourage multidimensional modeling of the pathophysiological role of 5-HT in BN. PMID- 14561434 TI - Cerebrospinal fluid tau levels in Alzheimer's disease are elevated when compared with vascular dementia but do not correlate with measures of cerebral atrophy. AB - Increased tau levels are a well-established finding in Alzheimer's disease (AD). In contrast, the potential value of tau levels in the differential diagnosis of AD, vascular dementia (VD) and major depression warrants further investigation. The potential impact of psychotropic medication also needs to be established. We investigated cerebrospinal fluid (CSF) tau protein concentrations in 88 patients with AD, 23 patients with VD, 25 patients with major depression and 17 age paralleled controls without cognitive impairment with respect to important clinical variables, type and dosage of psychotropic medication and cerebral changes as assessed by magnetic resonance imaging (MRI). The AD patients showed significantly elevated tau levels compared with patients with VD or major depression and controls. Tau levels obtained in the VD group were intermediate, with significant differences from both AD patients and patients with major depression and controls. Within the AD group, no significant correlation between tau levels, severity of dementia, age, duration of disease, type and dosage of psychotropic medication or MRI volumetric changes arose. A subgroup of AD patients without increased tau levels was characterized by a significantly larger percentage of patients with presenile onset. PMID- 14561435 TI - Serum cholesterol and impulsivity in a large sample of healthy young men. AB - Studies that have investigated the association between cholesterol levels and impulsivity are relatively few in number and have yielded equivocal results. In this study, we investigated the relationship between impulsivity, depression and serum lipids [total cholesterol, high-density lipoprotein (HDL) cholesterol, and triglycerides] in a large sample (N=2051) of healthy young men who were remarkably homogeneous in terms of age, educational level, and socioeconomic conditions. Depression was assessed using the depression scale of the Minnesota Multiphasic Personality Inventory-2, and impulsivity was measured using the impulse control scale of the Big Five Questionnaire (BFQ). We found that subjects with a low serum cholesterol, defined as the lowest tenth of the total cholesterol distribution (< or =3.7 mmol/l), scored significantly lower on the impulse control scale of the BFQ. There was no significant association between depression and cholesterol concentrations. In addition, in a multiple regression model, both lower levels of total cholesterol and higher levels of HDL cholesterol emerged as significant predictors of impulsivity. However, since the regression model accounted for only 0.6% of the variance in the score on the impulse control scale of the BFQ, the biological significance of these correlations was negligible. Taken together, these findings suggest that, in healthy young men, a relationship between cholesterol and impulsivity emerges only when the statistical analysis focuses on subjects with very low levels of cholesterol. PMID- 14561436 TI - Suicidal ideation and aggressiveness in school-aged youths. AB - This study set out to explore the frequency and correlates of suicidal ideation in a community sample of school-aged adolescents living in a northeastern area of Italy. The study involved a mixed male-female sample of 1,000 adolescents, corresponding to 10% of the young population aged 15-19 in the district. We investigated psychological distress through the revised Symptom Checklist (SCL-90 R), from which we also derived our indicators on suicidality (items concerning hopelessness and suicidal ideation). Propensity to aggression was measured with an Italian version of the Aggression Questionnaire. Females scored significantly higher than males on all SCL-90-R subscales, with the exception of the hostility subscale. In our sample, 30.8% of females and 25.3% of males reported having thought about suicide; one adolescent out of 20 reported suicidal ideation of high intensity (5% out of the total sample in both genders). Males scored significantly higher than females on the Aggression Questionnaire. Both males and females who reported suicidal feelings had significantly higher scores on the Aggression Questionnaire than did those who denied suicidal ideation. Suicidal ideation is a rather widespread occurrence among adolescents and correlates with other indicators of psychological distress. Psychological distress in adolescence might also be expressed through turbulent behaviors and conduct disorders. Identification of the mental disorders underlying a higher propensity to aggression might help to prevent other negative outcomes such as suicide. PMID- 14561437 TI - Influence of mirtazapine on urinary free cortisol excretion in depressed patients. AB - Mirtazapine has been shown to acutely inhibit cortisol secretion in healthy subjects. In the present study, the impact of mirtazapine treatment on urinary free cortisol (UFC) excretion was investigated in depression. Twenty patients (six men, 14 women) suffering from major depression according to DSM-IV criteria were treated with mirtazapine for 3 weeks. The patients received 15 mg mirtazapine on day 0; 30 mg mirtazapine on day 1; and 45 mg mirtazapine per day from day 2 to the end of the study (day 21). UFC excretion was measured before treatment (day 1), at the beginning (day 0), after 1 week (day 7) and after 3 weeks (day 21) of treatment with mirtazapine. Urine samples were collected from 08:00 to 08:00 h the following day. On the days of urine sampling, the severity of depressive symptoms was assessed using the 21-item version of the Hamilton Rating Scale for Depression (21-HAMD). There was a significant reduction of UFC excretion during 3-week mirtazapine therapy, which was already obvious after the first day of treatment (day 0). However, there were no significant across subjects correlations between UFC reduction and decrease in 21-HAMD sum scores. Apparently, the mirtazapine-induced rapid reduction of cortisol secretion in depressed patients is not necessarily correlated with a favorable therapeutic response. PMID- 14561438 TI - Reduced response-inhibition in obsessive-compulsive disorder measured with topographic evoked potential mapping. AB - Recent neuroimaging studies have suggested that a hyperactivity of the frontal striate neuronal circuits, including the orbitofrontal cortex and the basal ganglia, mediates the symptomatology of obsessive-compulsive disorder (OCD). However, there is also some evidence that the superior frontal cortex is less activated in OCD, and this local hypoactivity has been shown to be negatively associated with the symptomatology. As the superior frontal cortex is believed to be involved in inhibitory control, this study investigated the brain electrical activity during response inhibition in OCD. Twelve patients with OCD and 12 healthy controls performed a cued Go-NoGo task (continuous performance test), while event-related potentials were registered with 21 electrodes. Patients reacted significantly faster than controls, but did not differ from controls regarding the error rate. As a main result, we found a reduced frontal activity during the NoGo condition in OCD, which was condensed in a reduced anteriorisation of the brain electrical field. We suggest that this inhibitory deficit in OCD has a major contribution to the pathophysiology of OCD, which is underscored by the fact that the anteriorisation during the NoGo condition (NGA) was negatively correlated with the symptomatology as measured by the Yale-Brown Obsessive-Compulsive Scale. PMID- 14561439 TI - Depression with racing thoughts. AB - The aim of this study was to explore the clinical and family history correlates of depression with racing thoughts, an understudied phenomenon. Consecutive outpatients with a major depressive episode (MDE, N=336; unipolar subtype, n=130; bipolar-II subtype, n=206) were interviewed with the Structured Clinical Interview for DSM-IV Axis I Disorders-Clinician Version. Depression with racing thoughts was present in 213 patients (63.3%), a subgroup characterized by significantly more patients with bipolar-II disorder, lower age and lower age of onset, more atypical features, psychomotor agitation, diminished ability to think, suicidal ideation, guilt, leaden paralysis, MDE symptoms, and bipolar-II disorder family history than found in the subgroup without racing thoughts. Logistic regression controlled the diagnosis of bipolar-II disorder (which was associated with most of these variables). Comparisons in the separate bipolar-II and unipolar samples of depression with racing thoughts vs. the variables found significantly different in the total group found that associations with depression with racing thoughts were partly related to bipolar-II and partly related to unipolar diagnoses. Limitations of the study include reliance upon a single interviewer, non-blind cross-sectional assessment and bipolar-II diagnosis based on history. Depression with racing thoughts was very common in depressed outpatients, and was associated with suicidal ideation. Possible differential effects of antidepressants vs. mood stabilizers and antipsychotics are discussed. PMID- 14561440 TI - Respiratory panic disorder subtype: acute and long-term response to nortriptyline, a noradrenergic tricyclic antidepressant. AB - The goal of the study was to describe with prospective methodology the therapeutic response to nortriptyline in the respiratory panic disorder (PD) subtype versus the non-respiratory subtype. A total of 118 PD outpatients (DSM IV) were previously divided into respiratory (n=77) and non-respiratory (n=41) subtypes and then treated with nortriptyline for 1 year. Demographic and clinical features were compared in the two groups. Anxiety scales were administered before and during the treatment by raters who were blind to the subtype diagnosis. The principal instruments used to evaluate response were the Clinical Global Impression, the Sheehan Panic and Anticipatory Anxiety Scale, and the Panic Disorder Severity Scale. In the first 8 weeks of treatment (acute phase), the respiratory subtype had a significantly faster response on all the major scales. At the end of the study (week 52), there was no difference in the scale scores, and the reduction in panic attacks from baseline to end-point did not differ significantly between the two groups. In the respiratory subtype, the disorder had a later onset, was associated with a high familial history of mental disorder, and significantly more often required treatment with more than an occasional benzodiazepine. The non-respiratory subtype had significantly more previous depressive episodes. In conclusion, the respiratory PD subtype had a faster response to treatment with nortriptyline at 8 weeks than did the non respiratory subtype, and an equivalent response after 1 year of treatment. PMID- 14561442 TI - The neural substrate of analogical reasoning: an fMRI study. AB - This study investigated the anatomical substrate of analogical reasoning using functional magnetic resonance imaging. In the study, subjects performed a verbal analogy task (e.g., soldier is to army as drummer is to band) and, to control for activation caused by purely semantic access, a semantic judgment task. Significant activation differences between the verbal analogy and the semantic judgment task were found bilaterally in the prefrontal cortex (right BA 11/BA 47 and left BA45), the fusiform gyrus, and the basal ganglia; left lateralized in the postero-superior temporal gyrus (BA 22) and the (para) hippocampal region; and right lateralized in the anterior cingulate. The role of these areas in analogical reasoning is discussed. PMID- 14561443 TI - Electrophysiological comparison of grammatical processing and semantic processing of single spoken nouns. AB - Access to meaning from speech input is a high-speed process. However, models of word comprehension stipulate that words must be analyzed at phonological, grammatical and semantic levels. Here we used event-related potentials (ERPs) to compare on-line semantic categorization (natural/manufactured) and grammatical gender categorization (masculine/feminine) of spoken monosyllabic French noun pairs. Twenty four native French speakers were instructed to spot pairs of nouns in which both stimuli pertained to a category specified prior to each block. They could make a decision either after processing the first noun (i.e., ignore the second, Release condition), or after processing both (Hold condition). Virtually identical N4 components affecting Hold and Release ERPs in relation to category expectations were elicited by the first noun in both the tasks. Their topography was intermediary between that of the N400 and that of the Left Anterior Negativity. Despite shorter Release reaction times (RTs) for gender decisions than semantic ones, Release and Hold ERPs diverged 84 ms earlier in the semantic context than in the gender context, indicating faster onset for the processing of meaning. Conversely, the offset of Hold/Release differences was observed 42 ms earlier in the gender task than in the semantic task, suggesting an earlier completion for gender categorization. In sum, electrophysiological differences induced by semantic operations commenced earlier but recovered later than those relating to grammatical gender analysis. In convergence with previous Lateralized Readiness Potential and N200 studies, our results suggest that conscious semantic access can precede access to other types of lexical information during word comprehension in highly controlled conditions. PMID- 14561444 TI - Lexical processing in Spanish patients with probable Alzheimer's disease. AB - Twenty Spanish patients with probable Alzheimer's disease (AD) and 20 matched controls were given a battery of 17 tasks involving object recognition and the spoken and written perception and production of words and non-words. The AD patients were significantly impaired on nine of the tasks. Prominent among these were tasks that involve semantic processing, though non-word reading was also impaired. Performance on a category fluency task best discriminated AD patients from controls. It is proposed that impairment to semantic processing underlies most of the observed deficits on lexical processing tasks in patients with early AD, but that non-word reading may be sensitive to additional, mild impairments to phonological representations caused by extension of the degenerative process from anterior to posterior temporal regions. PMID- 14561445 TI - Spectrotemporal window of integration of auditory information in the human brain. AB - The human auditory system is adapted to integrate temporally successive sounds into meaningful entities, that is, acoustic information units. Hence, sound sequences falling within the temporal window of integration should be coded holistically as unitary representations in the human auditory cortex. Although it is well established that the auditory system operates in the frequency-temporal domain, many previous studies only focused on the temporal domain of the window of integration. Therefore, in the current study we investigated the relationship between the short-term temporal integration and the frequency integration. Event related magnetic fields in response to infrequent omission of the second tone in repetitive tone pairs composed of two closely spaced tones of different frequencies were recorded. This omission elicited the magnetic counterpart (MMNm) of the electric mismatch negativity (MMN), a change-specific component mainly generated in the auditory cortex, when the interval between the two successive tones was extremely short or when the frequency difference between the two tones was small. These findings suggest that two stimuli presented in close succession might be represented in the auditory system as a unitary integrated event. In addition, as the distance between the two successive tones decreased in the spectrotemporal dimensions, the magnitude of the MMNm increased. Behavioral data also supported these neurophysiological phenomena. This work shows the first neurophysiological evidence that the two-dimensional (spectrotemporal) window of integration, which provides important constraints for the neural processing of the acoustic environment, exists in the human brain. PMID- 14561446 TI - Dynamics of sensorimotor cortex activation to spatial sounds precueing ipsi- versus contralateral manual responses. AB - Spatially informative visual precues give rise to event-related potential asymmetries with higher negativities over the contralateral hemisphere. However the attribution of these potentials to sensorimotor areas is still unclear. The present magnetoencephalography study assessed movement preparation processes to auditory spatial precues. Event-related desynchronization (ERD) was measured to test the hypothesis that lateralized sounds would give rise to a fast, stimulus driven activation of motor networks independent of the precued response side. The lateralized vowels /a/ and /e/ served as precues for either ipsi- or contralateral responses, respectively, which had to be executed when an imperative stimulus was presented 1 s after precue onset. Two separate experiments were conducted with either blocked or mixed presentation of ipsi- and contralateral precues. Beta ERD over sensorimotor regions representing the stimulus side was elicited by both types of precues approximately 200 ms after their onset. For contralateral precues, a switch of beta ERD to the response hemisphere took place approximately 400 ms after trial-onset, peaking prior to the imperative stimulus (approximately 800 ms post trial-onset). Signal subspace projection demonstrated a high topographical correspondence between the early precue-related ERD and the pattern immediately preceding the response, suggesting that both were generated in similar motor networks. Apparently lateralized sounds give rise to an early activation of contralateral motor networks independent of the precued response. This suggests strong associations between space processing and action preparation networks, with fast activations preceding a detailed cortical analysis of stimulus meaning. PMID- 14561447 TI - Semantically convergent and semantically divergent priming in the cerebral hemispheres: lexical decision and semantic judgment. AB - The effects of semantically divergent and semantically convergent priming on the processing of lexical ambiguity by the two cerebral hemispheres were examined in two visual hemifield experiments. The experiments investigated the ability of the right hemisphere (RH) and the left hemisphere (LH) to summate activation from two single word primes followed by a laterally presented ambiguous target word. In a lexical decision task (experiment 1), the two priming words were either both related to the dominant meaning of the target (new, fresh-NOVEL), or to the subordinate meaning (story, book-NOVEL), or to one dominant and one subordinate meaning (new, story-NOVEL). Results indicated that the LH benefited most from semantically convergent primes that converged onto the dominant meaning of the ambiguous target word, whereas the RH benefited most from semantically mixed (divergent) primes, that diverge on alternate meanings of the ambiguous target word. We used the same stimuli in a semantic relatedness judgment task (experiment 2), and found that the facilitation in the RH was significantly larger when the primes were mixed than when both primes converged on a single (i.e. either dominant or subordinate) meaning of the to-be-presented target word. In contrast, the only facilitation found in the LH was when the two primes were associated with a single meaning (either dominant or subordinate) of the to-be presented target word. When the primes were mixed, there was no facilitation in the LH. These results support previous findings indicating that during word recognition, the RH activates a broader range of related meanings than the LH, including alternate meanings of ambiguous words. Thus, by summating activation for seemingly incongruous elements, the RH may be critically involved in at least one important component of verbal creativity. PMID- 14561448 TI - Neuromagnetic imaging of cortical oscillations accompanying tactile stimulation. AB - We applied a new method of imaging frequency-specific changes in brain activity in humans during a finger brushing task in order to measure changes in cortical rhythms during tactile stimulation. Neuromagnetic recordings were conducted in five subjects using a whole-head MEG system during tactile stimulation of the right index finger, with or without visual feedback, and while viewing another individual's index finger being stimulated. Volumetric images of changes in source power relative to pre-stimulus baseline levels were computed with 2 mm resolution over the entire brain using a minimum-variance beamforming algorithm (synthetic aperture magnetometry). Onset of tactile stimulation produced a brief (200-300 ms) suppression of mu band (8-15 Hz) and beta band (15-30 Hz) cortical activity in the primary somatosensory and primary motor cortex, respectively, followed by a bilateral increase in beta band activity ('beta rebound') in motor cortex. This pattern of suppression/rebound was absent when subjects observed finger brushing or brushing motions without receiving stimulation. In contrast, these conditions resulted in bilateral increases in beta band activity in sensorimotor areas and decreased power in the alpha (8-12 Hz) band in primary visual areas. These results show that spatially filtered MEG provides a useful method for directly imaging the temporal sequence of changes in cortical rhythms during transient tactile stimulation, and provide evidence that observation of tactile input to another individual's hand, or object motion itself, can influence independent rhythmic activity in visual and sensorimotor cortex. PMID- 14561449 TI - Early processing of the six basic facial emotional expressions. AB - Facial emotions represent an important part of non-verbal communication used in everyday life. Recent studies on emotional processing have implicated differing brain regions for different emotions, but little has been determined on the timing of this processing. Here we presented a large number of unfamiliar faces expressing the six basic emotions, plus neutral faces, to 26 young adults while recording event-related potentials (ERPs). Subjects were naive with respect to the specific questions investigated; it was an implicit emotional task. ERPs showed global effects of emotion from 90 ms (P1), while latency and amplitude differences among emotional expressions were seen from 140 ms (N170 component). Positive emotions evoked N170 significantly earlier than negative emotions and the amplitude of N170 evoked by fearful faces was larger than neutral or surprised faces. At longer latencies (330-420 ms) at fronto-central sites, we also found a different pattern of effects among emotions. Localization analyses confirmed the superior and middle-temporal regions for early processing of facial expressions; the negative emotions elicited later, distinctive activations. The data support a model of automatic, rapid processing of emotional expressions. PMID- 14561450 TI - Electrophysiological evidence for serial sentence processing: a comparison between non-preferred and ungrammatical continuations. AB - Event-related potentials (ERPs) were used to address a much debated issue in sentence processing research, namely whether one or multiple interpretations are pursued in case of syntactic ambiguities. For example in the syntactically ambiguous fragment "The man is painting the house and the garage...", 'and' either connects 'the house' and 'the garage', or conjoins two clauses (e.g., "The man is painting the house and the garage is already finished"). According to serial models, only one syntactic interpretation (the simplest) is pursued first (the first interpretation in the example). If this interpretation is incompatible with subsequently incoming words, the syntactic analysis of the preceding sentence fragment is revised. In contrast, parallel models propose that multiple interpretations of the ambiguity are pursued simultaneously. The two models were tested by comparing ERPs to words that were either ungrammatical, or grammatical but non-preferred continuations of the preceding sentence fragment. In a serial model, these two are not distinguished until after initial revision; in a parallel model, a distinction can be made at an early stage. The results supported a serial model: both with an acceptability judgment and a passive reading task a left lateralized negativity was found for ungrammatical and non preferred continuations relative to unambiguous grammatical continuations, which indicates that ungrammatical and non-preferred continuations were initially processed in the same way. However, in later time intervals, the ERPs to the ungrammatical continuation showed a posterior positivity (P600), whereas the ERPs to the non-preferred continuation had a more anterior focus, which indicates that they were processed differently. PMID- 14561451 TI - Localization of the event-related potential novelty response as defined by principal components analysis. AB - Recent research indicates that novel stimuli elicit at least two distinct components, the Novelty P3 and the P300. The P300 is thought to be elicited when a context updating mechanism is activated by a wide class of deviant events. The functional significance of the Novelty P3 is uncertain. Identification of the generator sources of the two components could provide additional information about their functional significance. Previous localization efforts have yielded conflicting results. The present report demonstrates that the use of principal components analysis (PCA) results in better convergence with knowledge about functional neuroanatomy than did previous localization efforts. The results are also more convincing than that obtained by two alternative methods, MUSIC-RAP and the Minimum Norm. Source modeling on 129-channel data with BESA and BrainVoyager suggests the P300 has sources in the temporal-parietal junction whereas the Novelty P3 has sources in the anterior cingulate. PMID- 14561452 TI - Functional anatomy of impaired selective attention and compensatory processing in autism. AB - In autism, physiological indices of selective attention have been shown to be abnormal even in situations where behaviour is intact. This divergence between behaviour and physiology suggests the action of some compensatory process of attention, one which may hold clues to the aetiology of autism's characteristic cognitive phenotype. Six subjects with autism spectrum disorders and six normal control subjects were studied with functional magnetic resonance imaging while performing a bilateral visual spatial attention task. In normal subjects, the task evoked activation in a network of cortical regions including the superior parietal lobe (P<0.001), left middle temporal gyrus (P=0.002), left inferior (P<0.001) and middle (P<0.02) frontal gyri, and medial frontal gyrus (P<0.02). Autistic subjects, in contrast, showed activation in the bilateral ventral occipital cortex (P<0.03) and striate cortex (P<0.05). Within the task condition, a region-of-interest comparison of attend-left versus attend-right conditions indicated that modulation of activation in the autistic brain as a function of the lateral focus of spatial attention was abnormally decreased in the left ventral occipital cortex (P<0.03), abnormally increased in the left intraparietal sulcus (P<0.01), and abnormally variable in the superior parietal lobe (P<0.03). These results are discussed in terms of a model of autism in which a pervasive defect of neural and synaptic development produces over-connected neural systems prone to noise and crosstalk, resulting in hyper-arousal and reduced selectivity. These low-level attentional traits may be the developmental basis for higher order cognitive styles such as weak central coherence. PMID- 14561453 TI - Speech and non-speech processing in hemispherectomised children: an event-related potential study. AB - Although hemispherectomy is now used as a radical treatment for intractable seizures in a number of centres, there have been limited electrophysiological studies investigating post-procedure auditory-speech processing and recovery or reorganisation. We therefore recorded auditory event-related potentials (ERPs) evoked by pure tones and syllables employing a 51-channel electrode array concentrated over the functional hemisphere in 17 patients (nine males, mean age 14.2 years) who had undergone hemispherectomy for intractable seizures; eight of the patients had congenital brain damage and nine had sustained their initial insult at an age of 1 year or older. For comparison, recordings were made from 10 controls (five males, mean age 13.5 years). Responses from patients consisted of five small amplitude components restricted to the functional hemisphere whose scalp polarity was opposite to the ERPs observed in control subjects. The topography of the auditory ERPs in the patients was localised to the centro temporal regions of the functional hemisphere in comparison to the midline distribution observed in control subjects. As in the control subjects, the latencies of the components evoked by tones were shorter than those evoked by syllables in both left and right hemispherectomised children. In addition, further analysis of the N1 and P2 component revealed increased syllable latencies in left hemispherectomised children only. The results of this study suggest that the left hemisphere alone may be more efficient at processing both pure tones and syllables compared to the right or both together. PMID- 14561454 TI - The Tower of London: the impact of instructions, cueing, and learning on planning abilities. AB - The Tower of London (ToL) is a well-known test of planning ability, and commonly used for the purpose of neuropsychological assessment and cognitive research. Its widespread application has led to numerous versions differing in a number of respects. The present study addressed the question whether differences in instruction, cueing, and learning processes systematically influence ToL performance across five difficulty levels (three to seven moves). A total of 81 normal adults were examined in a mixed design with the between-subject factor instruction (online versus mental preplanning) and the within-subject factors cueing (cue versus non-cue test version) and learning processes (first block and second block). We also assessed general intelligence for further analyses of differences between instruction groups. In general, there was a significant main effect across the difficulty levels indicating that the rate of incorrect solutions increased with problem difficulty. The participants who were instructed to make full mental plans before beginning to execute movements (preplanning) solved significantly more problems than people who started immediately with task related movements (online). As for the cueing conditions, participants with the minimum number of moves predetermined (cue) could solve more trials than people who were only instructed to solve the problems in as few moves as possible (non cue). Participants generally increased performance in the second part of the test session. However, an interaction of presentation order of the cueing condition with learning indicated that people who started the tasks with the non-cue version showed significantly better performance in the following cue condition, while participants who started with the cue condition stayed at the same performance level for both versions. These findings suggest that instruction, cueing conditions, and learning processes are important determinants of ToL performance, and they stress the necessity of standardized application in research and clinical practice. PMID- 14561455 TI - Mismatch negativity indexes auditory temporal resolution: evidence from event related potential (ERP) and event-related field (ERF) recordings. AB - This study examined auditory temporal resolution as indexed by gap detection using the mismatch negativity (MMN) component of the auditory event-related potential (ERP) and its magnetic counterpart (MMNm). ERPs were recorded in 10 subjects who were presented with auditory stimuli. These stimuli were presented in sequences of repetitive continuous 'standard' sinusoidal tones interspersed with infrequently occurring 'deviant' stimuli that differed from standards only in that they contained a silent gap midway in the stimulus. The gap size varied in separate stimulus blocks and was either 3, 5 or 7 ms. The stimuli were presented monaurally either to the left or the right ear. In a separate session, event-related magnetic fields (ERFs) were recorded from eight subjects using a similar paradigm but with gap sizes of 3, 7 or 11 ms and with binaural stimulation. Both ERP and ERF recordings showed that the smallest gap size (3 ms) did not elicit as large or reliable MMN or MMNm as did the larger ones. There were no differences in the laterality of the MMN as might be predicted on the basis of previous behavioural studies, but this result is likely a reflection of differences in task requirements. Nonetheless, the findings suggest that MMN and MMNm successfully index auditory temporal resolution thresholds, as measures that are independent of attention. PMID- 14561456 TI - Identification of prosodic attitudes by a temporal recurrent network. AB - Human speakers modulate the fundamental frequency (F0) of their utterances in order to express different 'prosodic' attitudes such as surprise or curiosity. How are these prosodic attitudes then decoded? The current research addresses the issue of how the temporal structure of F0 can be used in order to discriminate between prosodic attitudes in natural language using a temporal recurrent neural network (TRN) that was initially developed to simulate the neurophysiology of the primate frontostriatal system. In the TRN, a recurrent network of leaky integrator neurons encodes a continuous trajectory of internal states that characterizes the input sequence. The input to the model is a population coding of the continuous, time-varying values of the fundamental frequency (F0) of natural language sentences. We expose the model to an experiment based on one in which human subjects were required to discriminate between different prosodic attitudes (surprise, exclamation, question, etc.). After training, the model discriminates between six prosodic attitudes in new sentences at 82.52% correct, compared to 72.8% correct for human subjects. These results reveal (1) that F0 provides relevant information for prosodic attitude discrimination, and (2) that the TRN demonstrates a categorical sensitivity to this information that can be used for classifying new sentences. PMID- 14561457 TI - When and how do listeners relate a sentence to the wider discourse? Evidence from the N400 effect. AB - In two ERP experiments, we assessed the impact of discourse-level information on the processing of an unfolding spoken sentence. Subjects listened to sentences like Jane told her brother that he was exceptionally quick/slow, designed such that the alternative critical words were equally acceptable within the local sentence context. In Experiment 1, these sentences were embedded in a discourse that rendered one of the critical words anomalous (e.g. because Jane's brother had in fact done something very quickly). Relative to the coherent alternative, these discourse-anomalous words elicited a standard N400 effect that started at 150-200 ms after acoustic word onset. Furthermore, when the same sentences were heard in isolation in Experiment 2, the N400 effect disappeared. The results demonstrate that our listeners related the unfolding spoken words to the wider discourse extremely rapidly, after having heard the first two or three phonemes only, and in many cases well before the end of the word. In addition, the identical nature of discourse- and sentence-dependent N400 effects suggests that from the perspective of the word-elicited comprehension process indexed by the N400, the interpretive context delineated by a single unfolding sentence and a larger discourse is functionally identical. PMID- 14561458 TI - High density ERP indices of conscious and unconscious semantic priming. AB - The existence of differential brain mechanisms of conscious and unconscious processing is a matter of debate nowadays. The present experiment explores whether conscious and unconscious semantic priming in a lexical decision task at a long prime-target stimulus onset asynchrony (SOA) correlate with overlapping or different event related potential (ERP) effects. Results show that the N400 effect, which appeared when words were consciously perceived, completely disappeared when primes were masked at a level where the ability of participants to detect the prime was near chance. Instead, a rather different set of ERP effects was found to index unconscious semantic priming. This suggests that the processes at the basis of conscious and unconscious semantic analyses can under some circumstances be rather different. Moreover, our results support the notion that conscious and unconscious processes are at least partially separable in the brain. PMID- 14561459 TI - Functional neuroanatomy of interference in overlapping dual tasks: an fMRI study. AB - A basic characteristic of the human action and cognition system is the occurrence of interference when participants attempt to perform two tasks at the same time. Such interference has been studied for a long time with so-called overlapping dual tasks, where two stimuli presented in rapid succession require separate responses. As an indicator of interference, reaction times on the second stimulus increase the smaller the interval between both tasks. While most behavioral studies investigated the temporal dynamics of the interference, we focused on the functional neuroanatomy of overlapping dual-task performance by using functional magnetic resonance imaging (fMRI). Participants were asked to perform two choice reaction tasks concurrently [Pashler, Psychol. Bull., 116 (1994) 220-244]. When activation in this overlapping dual-task situation was compared with the summed activation of the single component tasks, activation in the prefrontal, temporal, parietal, and occipital cortices was detected. These data suggest that the processing of the overlapping dual tasks requires an extensive and distributed network of processing centers. However, the main focus of the dual-task-related activation was located in regions surrounding the left inferior frontal sulcus. Based on our findings and on findings of other recent neuroimaging studies, we argue that activation of the left inferior frontal sulcus reflects increased synaptic activity related to the need to manage interfering information in order to determine the appropriate action. PMID- 14561460 TI - Altered auditory processing in acutely psychotic never-medicated first-episode patients. AB - Individuals with psychosis fail to differentiate external impulses and suffer from distortions of reality testing. Schizophrenia group illnesses are also associated with deficits in working memory and perception. We examined the manifestations of a very early phase of psychotic illness to automatic auditory deviance detection to clarify the basic mechanisms underlying misinterpretations of perception. METHODS: Twenty-five never-medicated patients admitted for hospital evaluation of acute psychosis were studied. Fifty-eight EEG channels were recorded during an auditory oddball paradigm. Event-related potentials (ERPs) time-locked to non-attended deviant auditory stimuli were studied in patients and compared with healthy controls. Auditory processing was examined both at the level of the measured biosignals (standard and deviant responses) and with subtraction waveforms. Topographical differences were characterized using global field power (GFP) and minimum norm estimates. RESULTS: The maximum GFP amplitudes and mean amplitudes of the 58 channels within the time windows corresponding to the previously known 'N2b', 'P3a' and 'P3b' components were clearly reduced in patients when compared to healthy controls. However, the groups did not differ during attention-independent automatic processing corresponding to the 'N1' and 'MMN' components, or with respect to the peak latencies of the GFP maxima. CONCLUSIONS: Impairment of the processing of a deviance in simple auditory input in acutely ill drug-naive first-episode psychotic patients only appears in attention-dependent processing after about 250 ms. The alterations in auditory processing differed between stimulus types, suggesting at least two mechanisms underlying the auditory discrimination impairments in acute psychosis. After 250 ms there was a linear and gradually increasing difference in magnitude between the groups in their responses to deviant stimuli, probably related to arousal. In addition, however, there was a striking difference between the groups in the processing of standard stimuli. The early processing was similar in patients and controls, but the striking difference appeared in later processing. The sensory memory deficits associated with psychosis may be explained by an abnormality in sensory model formation rather than by impaired deviant detection. PMID- 14561462 TI - Perceptual dominance of oriented faces mirrors the distribution of orientation tunings in inferotemporal neurons. AB - In three experiments participants viewed pairs of overlapping transparent faces, with one face upright and the other oriented, and they reported which face was dominant. In each trial, an upright face was presented with a face at 45, 90, 135 or 180 degrees, with transparency set using a linear weighted algorithm, so that relative contrast across faces was biased in favour of oriented faces. Exposure duration was restricted in experiment 1 to 250, 500 or 1000 ms, but was unlimited in experiments 2 and 3. Adults were tested in experiments 1 and 2 and children aged 6-9 years of age were tested in experiment 3. Irrespective of exposure duration, the results showed the probability of dominance being ceded by oriented faces to upright faces was a function of orientation. In comparable conditions, the function found with young children was flatter than with adults. These patterns, and those of earlier perceptual studies, can be explained by the distribution of different orientation tunings found in physiological studies of inferotemporal cortex in macaques. PMID- 14561461 TI - The volitional inhibition of anticipatory ocular pursuit using a stop signal. AB - Unlike limb movements, smooth pursuit eye movements cannot normally be performed in the absence of a target. However, when subjects have a high expectancy of an imminent target appearance, the situation changes, and anticipatory smooth pursuit (ASP) tends to precede target onset by several hundred milliseconds. The velocity of this ASP is scaled predictively according to expected target velocity. And when an upcoming target is unexpectedly altered, or fails to appear, ASP continues regardless for approximately 150-200 ms before modification by visual feedback begins [J. Neurophysiol., 84 (2000) 2340]. These and other observations led to the earlier suggestion that ASP might be ballistic, being pre programmed from start to finish. Two experiments with different timing parameters were therefore performed to test this hypothesis using a version of Logan's [Psychol. Rev., 91 (1984) 295] stop signal task. The aim was to test whether ASP could be stopped at will, and if so, whether the time taken to stop varied as a function of the time since ASP onset. Results showed that in response to a stop signal, ASP can be inhibited at any point in its trajectory, and for the majority of subjects in experiment 1, and all the subjects in experiment 2, with a latency that does not change significantly with target speed or time since ASP onset. These results provide the first demonstration that anticipatory movements can be stopped volitionally in response to a stop signal. Possible cognitive and neurophysiological mechanisms underlying this process are discussed. PMID- 14561463 TI - Perceptual learning modulates sensory evoked response during vowel segregation. AB - With practice, people become better at discriminating two similar stimuli, such as two sounds. The neural mechanisms that underlie this type of learning have been of interest to researchers investigating neural plasticity associated with learning and recovery of function following stroke. We utilized event related potentials (ERP) to study the neural substrates underlying auditory discrimination learning. Stimuli were five steady-state American English vowels. On each trial, participants were presented with a pair of vowels created by summing together the digital waveforms of two different vowels. Listeners were instructed to identify both vowels in the pair. ERPs were recorded during two sessions separated by 1 week. Half of the participants practised the discrimination task during the intervening week while the other half served as controls and did not receive any training. Trained listeners showed greater improvement in accuracy than untrained participants. In both groups, vowels generated N1 and P2 waves at the fronto-central and temporal scalp regions. The behavioral effects of training were paralleled by decreased N1 and P2 latencies as well as enhanced P2 amplitude in the trained compared with untrained listeners. The effects of training on sensory evoked responses are consistent with the proposal that perceptual learning is associated with changes in sensory cortices. PMID- 14561464 TI - Neural correlates of error detection and correction in a semantic retrieval task. AB - Event-related potentials (ERPs) were used to investigate the cognitive and neural substrates of immediate and 1-week delayed error correction in a semantic retrieval task. In particular, we pursued the basis for the 'hypercorrection' effect, the finding that erroneous responses endorsed as correct with high confidence are more likely than low-confidence errors to be corrected at retest. Presentation of negative, but not positive feedback about the accuracy of one's response elicited a fronto-central negativity, similar to the ERN, which was somewhat sensitive to the degree to which negative feedback violated expectation. A fronto-central positivity, similar to the novelty-P3/P3a, more generally indexed detection of a metamemory error, given that it was larger in conditions of high metamemory mismatch than in conditions of low metamemory mismatch, irrespective of absolute task accuracy. For errors, amplitude of the fronto central positivity, but not the preceding negativity, was correlated with correction on an immediate retest. Thus, to the extent that the fronto-central positivity indexes an orienting response, this response appears to facilitate initial encoding processes, but does not play a key role in memory consolidation. In contrast, a broad, inferior-temporal negativity occurring 300-600 ms after presentation of the correct answer was sensitive to subsequent memory performance at both immediate and delayed retests, but only for answers containing familiar semantic information. This negativity may reflect processes involved in the formation of an association between the question and pre-existing semantic information. PMID- 14561465 TI - Tracking the time course of phonological encoding in speech production: an event related brain potential study. AB - This study investigated the time course of phonological encoding during speech production planning. Previous research has shown that conceptual/semantic information precedes syntactic information in the planning of speech production and that syntactic information is available earlier than phonological information. Here, we studied the relative time courses of the two different processes within phonological encoding, i.e. metrical encoding and syllabification. According to one prominent theory of language production, metrical encoding involves the retrieval of the stress pattern of a word, while syllabification is carried out to construct the syllabic structure of a word. However, the relative timing of these two processes is underspecified in the theory. We employed an implicit picture naming task and recorded event-related brain potentials to obtain fine-grained temporal information about metrical encoding and syllabification. Results revealed that both tasks generated effects that fall within the time window of phonological encoding. However, there was no timing difference between the two effects, suggesting that they occur approximately at the same time. PMID- 14561466 TI - Language context and phonetic change detection. AB - Event-related brain potentials (ERP) were recorded to two spoken words, /paeti/ and /peti/. The vowel difference between the two words results in a semantical difference in Finnish, but not in Hungarian, in which /ae/ and /e/ are perceived as allophones of the same vowel /epsilon/. As a consequence, native Hungarian speakers, who had not studied Finnish before being tested, could not categorize the two word stimuli. In the main experiment, native Hungarian speakers, who fluently spoke Finnish, were presented with two oddball sequences in which /paeti/ was the frequent standard stimulus, /peti/ the infrequent deviant. In addition, very rare target words were also included. In one condition, the targets were Hungarian words, whereas in the other, they were Finnish words. The participants' sense of being in two different language environments was further encouraged by having separate experimenters conducting the two conditions, one speaking with the subjects only in Hungarian, the other only in Finnish. Language context had no effect on the mismatch negativity ERP component elicited by the deviant word stimuli. This result suggests that language context does not affect the pre-attentive detection of auditory deviance. PMID- 14561467 TI - Involvement of a novel fast inward sodium current in the invasion capacity of a breast cancer cell line. AB - This work reports the finding of a unique fast inward sodium current (I(Na)) in MDA-MB-231 cells which is missing in MDA-MB-468 cells and in MCF-7 cells. This current is high-voltage-activated and displays a window current at the membrane potential of MDA-MB-231 cells. This current is blocked by high concentrations of tetrodotoxin (TTX). In MDA-MB-231 cells, which are the most invasive cells among the three cell lines tested, proliferation and migration were not sensitive to TTX while invasion was reduced by approximately 30%. These experiments suggest that I(Na) is involved in the invasion process, probably through its participation to the regulation of the intracellular sodium homeostasis. PMID- 14561468 TI - Monoterpenes affect chlorodiazepoxide-micelle interaction through micellar dipole potential modifications. AB - The ability of several natural terpenes to affect benzodiazepine (BZD)-micelle interaction through the membrane dipolar organization was investigated. The acid base equilibrium of chlorodiazepoxide (CDX) and the spectroscopic behavior of the electrochromic dye merocyanine were tested in the presence and in the absence of Triton X-100 micelles (used to mimic a membrane environment) containing or not cineole, menthol, geraniol or camphor. CDX's apparent pK increased in the environment of terpene-containing micelles compared with pure Triton X-100 micelles. Decrements in electric potentials (between -111 and -128 mV with respect to pure detergent) were calculated from Boltzmann equation. This result suggested, that in the presence of terpenes, the tendency of CDXH(+) to remain in the membrane phase increased. The dielectric constant (D) of the microenvironment sensed by merocyanine within Triton X-100 micelles, determined from lambda(max,2) of merocyanine monomer, was D=9 and increased in the presence of all the terpenes assayed (D congruent with 11). The decrease in merocyanine partitioning (A(peak1)/A(peak2) increased) also reflected an increment in the negative dipole potential. The present results suggest that terpenes contributed to the whole dipolar arrangement of the micelle with a dipole moment vector which had an intense component oriented parallel to the intrinsic dipole of the Triton X-100 molecules in the micelles. This led to a more negative environment of the interface region where CDX was located, and increased the net polarity of the deepest micelle regions sensed by merocyanine. PMID- 14561469 TI - Molecular models of the intercellular lipid lamellae from epidermal stratum corneum. AB - The purpose of the present study was to test the hypothesis that the 13 nm trilamellar repeat units within the intercellular spaces of epidermal stratum corneum are composed of lamellae with alternating 5-3-5 nm dimensions as presented in previous models [J. Invest. Dermatol. 92 (1989) 251, P.W. Wertz, Integral lipids in hair and stratum corneum, in: P. Jolles, H. Zahn, H. Hocker (Eds.), Hair: Biology And Structure, Birkhauser Verlag, Basel, 1996, pp. 227-238, Acta Derm.-Venereol., Suppl. 208 (2000) 23]. Electron density profiles were measured from transmission electron micrographs of porcine stratum corneum prepared using ruthenium tetroxide [J. Invest. Dermatol. 92 (1989) 251]. Center to-center distances of adjacent electron-dense bands as well as adjacent lucent bands were measured. Dense band center-to-center measurements were consistent with a 5-3-5 nm arrangement. However, lucent band center-to-center measurements revealed uniform lamellar thickness. It is suggested that linoleate chains in the central lamella reduce more ruthenium than the predominantly saturated chains in the outer lamellae and that this additional reduced ruthenium accumulates under the polar head group regions. A similar phenomenon involving the sphingosine moieties of the covalently bound omega-hydroxyceramide molecules accounts for the three-band pattern seen between the ends of adjacent corneocytes. It is concluded that the component lamellae of the several types of 13 nm trilamellar units of the stratum corneum are all of equal thickness. PMID- 14561470 TI - Backbone dynamics of membrane proteins in lipid bilayers: the effect of two dimensional array formation as revealed by site-directed solid-state 13C NMR studies on [3-13C]Ala- and [1-13C]Val-labeled bacteriorhodopsin. AB - We have recorded site-directed solid-state 13C NMR spectra of [3-13C]Ala- and [1 13C]Val-labeled bacteriorhodopsin (bR) as a typical membrane protein in lipid bilayers, to examine the effect of formation of two-dimensional (2D) lattice or array of the proteins toward backbone dynamics, to search the optimum condition to be able to record full 13C NMR signals from whole area of proteins. Well resolved 13C NMR signals were recorded for monomeric [3-13C]Ala-bR in egg phosphatidylcholine (PC) bilayer at ambient temperature, although several 13C NMR signals from the loops and transmembrane alpha-helices were still suppressed. This is because monomeric bR reconstituted into egg PC, dimyristoylphosphatidylcholine (DMPC) or dipalmytoylphosphatidylcholine (DPPC) bilayers undergoes conformational fluctuations with frequency in the order of 10(4)-10(5) Hz at ambient temperature, which is interfered with frequency of magic angle spinning or proton decoupling. It turned out, however, that the 13C NMR signals of purple membrane (PM) were almost fully recovered in gel phase lipids of DMPC or DPPC bilayers at around 0 degrees C. This finding is interpreted in terms of aggregation of bR in DMPC or DPPC bilayers to 2D hexagonal array in the presence of endogenous lipids at low temperature, resulting in favorable backbone dynamics for 13C NMR observation. It is therefore concluded that [3-13C]Ala-bR reconstituted in egg PC, DMPC or DPPC bilayers at ambient temperature, or [3-13C]Ala- and [1-13C]Val-bR at low temperature gave rise to well-resolved 13C NMR signals, although they are not always completely the same as those of 2D hexagonal lattice from PM. PMID- 14561471 TI - Plasmalogens protect unsaturated lipids against UV-induced oxidation in monolayer. AB - Oxidative stress results from the attack by free radicals of several cellular targets (proteins, DNA and lipids). The cell equilibrium is a direct consequence of the pro-/antioxidant balance. In order to understand the physiological processes involved in oxidative stress, we followed oxidation of unsaturated lipids using a biomimetic system: Langmuir monolayers. The oxidation mode chosen was UV-irradiation and the lipid model was a polyunsaturated phospholipid: 1,2 dilinoleoyl-sn-glycero-3-phosphocholine (DLPC). The monomolecular film technique was used to measure membrane rheology before and after UV-irradiation. We showed that the UV-irradiation of a DLPC monomolecular film led to a molecular area and surface elasticity modulus decrease that attests to the apparition of new molecular species at the air-water interface. The antioxidant effect of a synthetic plasmalogen (1-O-(1'-(Z)-hexadecenyl)-2-O-oleoyl-sn-glycero-3 phosphocholine or P(PLM)OPE) was tested on the oxidation of DLPC. Indeed, for about 25% mol P(PLM)OPE in mixed DLPC/P(PLM)OPE monolayers, a complete inhibition of the molecular area and the surface elasticity modulus decreases was observed in our experimental conditions. Lower P(PLM)OPE quantities delayed but did not prevent the DLPC oxidation in mixed monolayers. PMID- 14561472 TI - Synthetic copoly(Lys/Phe) and poly(Lys) translocate through lipid bilayer membranes. AB - Several membrane-transporting peptides (MTP) containing basic amino acid residues such as Lys and Arg that carry macromolecules such as DNA and proteins across cell plasma membranes by an unknown mechanism have been actively studied. On the basis of these results, we have been investigating the translocation ability of synthetic polypeptides, copoly(Lys/Phe) and poly(Lys), through negatively charged phospholipid (soybean phospholipid (SBPL)) bilayer membranes by zeta potential analysis, circular dichroism (CD) spectroscopy, fluorescence spectroscopy, an electrophysiology technique, and confocal laser scanning microscopy (CLSM). The binding of these polypeptides to the membrane, which is the first step for translocation across the membrane, resulted in the conformational transition of the polypeptide from a random coil form or helix-poor form to a helix-rich form. The fluorescence studies demonstrated that the time-dependent decrease in the fluorescence intensities of the FITC-labeled polypeptides bound to the SBPL liposome reflected translocation of the polypeptide across the lipid bilayer with the low dielectric constant. Both the rate constant and the efficiency of the polypeptide translocation across the lipid bilayer were greater for copoly(Lys/Phe) than for poly(Lys). These results suggest that the random incorporation of the hydrophobic Phe residue into the positively charged Lys chain results in a lowering of the potential barrier for passage of the polypeptide in the hydrophobic core portion of the lipid bilayer. We presented the first direct observation that the positively charged polypeptides, copoly(Lys/Phe) (MW: 41,500) and poly(Lys) (MW: 23,400), could translocate across the lipid bilayer membrane. PMID- 14561473 TI - Iron and magnesium exchange via the low affinity iron transporter in rabbit erythroid cells-exchange rates and the action of valinomycin, diethylstilbestrol and protein kinase inhibitors. AB - Evidence was presented previously that rabbit erythroid cells possess a low affinity Fe2+ transport system which operates via the Na+/Mg2+antiport [Biochim. Biophys. Acta 1282 (1996) 163]. This was investigated further by measurements of Mg2+ efflux as well as Fe2+ uptake by the cells and by examining the inhibitory effects of valinomycin, diethylstilbestrol (DES) and protein kinase inhibitors. Mg2+ efflux and Fe2+ uptake were measured using rabbit reticulocytes and mature erythrocytes incubated in isotonic KCl or NaCl solutions. Both processes were slower in mature cells than reticulocytes. Mg2+ efflux into KCl solution was much lower than into NaCl solution but was stimulated by addition of Fe2+ to the solution. The rate of Fe2+-stimulated Mg2+ efflux closely followed that of Fe2+ uptake in a one-to-one molar ratio. Valinomycin, DES and the protein kinase inhibitors all inhibited Fe2+ uptake from KCl solution. Valinomycin also inhibited Fe2+-stimulated Mg2+ efflux into KCl solution but markedly stimulated the efflux into NaCl. Maximal inhibition of Fe2+ uptake from KCl solution required the presence of K+, Rb+ or Cs+ ions with which valinomycin forms strong complexes. The results could not be explained on the basis of changes in cell membrane potential or cell volume. By contrast, the increase in Mg2+ efflux into NaCl solution produced by valinomycin was accompanied by cell shrinkage and production of a more negative membrane potential, either of which may be responsible for the effect. The inhibition produced by the protein kinase inhibitors indicate that phosphorylation of the transporter or an associated protein by protein tyrosine kinase is probably required to activate the transporter. PMID- 14561474 TI - Non-cooperative effects of lung surfactant proteins on early adsorption to an air/water interface. AB - Two small hydrophobic proteins, SP-B and SP-C, are responsible for rapid adsorption of pulmonary surfactant to the air/water interface. Despite their physiological importance, the number of protein molecules required to trigger an absorption event remains unknown. To investigate this issue, we varied the protein content of calf lung surfactant extract (CLSE) by dilution with protein depleted surfactant lipids (neutral and phospholipids, N&PL). Vesicles of a constant size and of composition ranging between 100% N&PL and 100% CLSE were generated by probe sonication. Their adsorption kinetics to an air/water interface were monitored at different temperatures using a Wilhelmy plate to measure surface tension. When plotted versus protein concentration, the adsorption rates during the initial change in surface tension exhibit a diphasic behavior, first increasing rapidly and linearly between 0% and 25% CLSE, and then more slowly at higher concentrations. Direct linearity at low protein content (0 5% CLSE ratio) was confirmed at 37 degrees C. These observations argue against cooperative behavior, for which the adsorption rate would first rise slowly with the protein content, and then increase suddenly once the critical number of proteins on each vesicle is reached. The apparent activation energy E(a) and the free energy of activation DeltaG(0)*, calculated from the temperature dependence of adsorption, further support the view that at least the early stages of protein induced surfactant adsorption proceeds through a sequence of events involving not several, but a single surfactant protein. PMID- 14561475 TI - Miscibility and phase behaviour of binary mixtures of N-palmitoylethanolamine and dipalmitoylphosphatidylcholine. AB - The content of N-acylethanolamines (NAEs) increases dramatically in cell membranes when the parent organism is subjected to injury or stress. This increase has been attributed to stress-combating mechanisms of the organism. In this study, a binary phase diagram of hydrated mixtures of N palmitoylethanolamine (NP-E)--an endogenous ligand for the peripheral cannabinoid receptor (CB-2)--with dipalmitoylphosphatidylcholine (DPPC) is established by high-sensitivity differential scanning calorimetry (DSC). The structures of the phases involved were determined by using 31P-NMR spectroscopy and low-angle X-ray scattering. DSC studies show that NP-E and DPPC mix well in the composition range DPPC/NP-E=100:0 to 40:60 (mol/mol). At higher contents of NP-E, phase separation is indicated by the presence of additional transitions in the thermograms. Characterization of the structures formed by the mixtures with 31P-NMR shows that, up to 80 mol% NP-E, DPPC remains in the lamellar phase. The low-angle X-ray diffraction data are also consistent with a lamellar gel-phase structure for DPPC/NP-E mixtures up to 60 mol% NP-E. Above 70 mol% NP-E, NP-E phase separates in the gel-phase region, while complete miscibility is observed in the fluid phase. These results provide a structural basis for understanding the membrane interactions of NAEs, which is necessary for understanding the mechanism of their putative stress-combating role in the parent organisms. PMID- 14561476 TI - Association of hydrophobically-modified poly(ethylene glycol) with fusogenic liposomes. AB - We present results on using cooperative interactions to shield liposomes by incorporating multiple hydrophobic anchoring sites on polyethylene glycol (PEG) polymers. The hydrophobically-modified PEGs (HMPEGs) are comb-graft polymers with strictly alternating monodisperse PEG blocks (M(w)=6, 12, or 35 kDa) bonded to C18 stearylamide hydrophobes. Cooperativity is varied by changing the degree of oligomerization at a constant ratio of PEG to stearylamide. Fusogenic liposomes prepared from N-C12-DOPE:DOPC 7:3 (mol:mol) were equilibrated with HMPEGs. Affinity for polymer association to liposomes increases with the degree of oligomerization; equilibrium constants (given as surface coverage per equilibrium concentration of free polymer) for 6 kDa PEG increased from 6.1+/-0.8 (mg/m(2))/(mg/ml) for 2.5 loops to 78.1+/-12.2 (mg/m(2))/(mg/ml) for 13 loops. In contrast, the equilibrium constant for distearoylphosphatidylethanolamine poly(ethylene glycol) (DSPE-PEG5k) was 0.4+/-0.1 (mg/m(2))/(mg/ml). The multi loop HMPEGs demonstrate higher levels of protection from complement binding than DSPE-PEG5k. Greater protection does not correlate with binding strength alone. The best shielding was by HMPEG6k-DP3 (with three 6 kDa PEG loops), suggesting that PEG chains with adequate surface mobility provide optimal protection from complement opsonization. Complement binding at 30 min and 12 h demonstrates that protection by multi-looped PEGs is constant whereas DSPE-PEG5k initially protects but presumably partitions off of the surface at longer times. PMID- 14561477 TI - Novel properties of cholesterol-dioleoylphosphatidylcholine mixtures. AB - We have studied the properties of mixtures of cholesterol with dioleoylphosphatidylcholine (DOPC), and with several other phospholipids, including 1-stearoyl-2-oleoylphosphatidylcholine (SOPC) and dioleoleoylphosphatidylserine (DOPS), as a function of cholesterol molar fraction and of temperature. Mixtures of DOPC with a cholesterol molar fraction of 0.4 or greater display polymorphic behavior. This polymorphism includes the formation of structures that give rise to isotropic peaks in 31P NMR at cholesterol molar fractions between 0.4 and 0.6, dependent on the thermal history of the sample. Cryo-electron microscopy studies demonstrate the formation of small globular aggregates that would contribute to a narrowing of the 31P NMR powder pattern. At molar fraction cholesterol 0.6 and higher and at temperatures above 70 degrees C, the mixtures with DOPC convert to the hexagonal phase. Lipid polymorphism is accompanied by the phase separation of cholesterol crystals in the anhydrous form and/or the monohydrate form. The crystals that are formed have substantially altered kinetics of hydration and dehydration, compared with both pure cholesterol monohydrate crystals and with crystals formed in the presence of the other phospholipids that do not form the hexagonal phase in the presence of cholesterol. This fact demonstrates that these cholesterol crystals are in intimate contact with the DOPC phospholipid and are not present as morphologically separate structures. PMID- 14561478 TI - Synthesis of new 2-acetyl and 2-benzoyl quinoxaline 1,4-di-N-oxide derivatives as anti-Mycobacterium tuberculosis agents. AB - A series of 2-acetyl and 2-benzoyl-6(7)-substituted quinoxaline 1,4-di-N-oxide derivatives were synthesized and evaluated for in vitro antituberculosis activity. The results show that 2-acetyl-3-methylquinoxaline 1,4-di-N-oxide derivatives with chlorine, methyl or methoxy group in position 7 of the benzene moiety (compounds 2, 4 and 6, respectively) and unsubstituted (3) have good antitubercular activity, exhibiting EC(90)/MIC values between 0.80 and 4.29. In conclusion, the potency, selectivity and low cytotoxicity of these compounds make them valid leads for synthesizing new compounds that possess better activity. PMID- 14561479 TI - New N6- or N(9)-hydroxyalkyl substituted 8-azaadenines or adenines as effective A1 adenosine receptor ligands. AB - In this paper we describe synthesis and biological assays of some A(1) ligands more water-soluble than the effective, but very lipophilic, 8-azaadenines and adenines discovered in the past and obtained introducing on N(6) or N(9) substituent a hydroxy group. Five of the new N(6)-hydroxyalkyl- and N(6) hydroxycycloalkyl-2-phenyl-9-benzyl-8-azaadenines showed very high affinity (Ki<40 nM) and selectivity for A(1) adenosine receptors. Among the 2-phenyl-9-(2 hydroxy-3-alkyl)-8-azaadenines or adenines prepared, the compounds with the higher A(1) affinity and selectivity resulted 2-phenyl-9-(2-hydroxy-3-propyl) N(6)-cyclopentyl- and cyclohexyl-8-azaadenine with Ki 2.2+/-0.2 nM and 2.8+/-0.3 nM respectively. From the point of view of water-solubility, 2-phenyl-9-(2 hydroxy-3-propyl)-8-azaadenine was the most interesting compound, having a CLogP of 1.066991 and a water-solubility of 1.2 mg mL(-1). PMID- 14561480 TI - New styryl sulfones as anticancer agents. AB - New styryl sulfone compounds have been synthesized and evaluated for their anti proliferative activity. Among the compounds synthesized, one compound (7k) has shown 51% tumor growth inhibition in mice implanted with HT-29 human carcinoma at 400 mg kg(-1) orally. PMID- 14561481 TI - In vitro and in vivo evaluations of THAM derived telomers bearing RGD and Ara-C for tumour neovasculature targeting. AB - As an approach to the development of specific drug delivery systems, a new class of low macromolecular carriers called 'telomers' endowed with an antitumour agent, such as arabinofuranosylcytosine (Ara-C), RGDSK peptidic sequences, as tumour targeting moieties, and tyrosine groups labelled with 125I atoms allowing the in vivo scintigraphic follow up, were synthesized. Their tumour targeting ability was assessed in vivo in mice bearing a murine B16 melanoma. The biological results showed that the presence of RGDSK sequences onto the macromolecules leads to the selective targeting and the accumulation of telomers within the vascularized zone of the tumour. Moreover, such compounds exhibited in vitro a better IC(50) (0.015 muM) than pure Ara-C and in vivo an oncostatic index higher than 160%. PMID- 14561482 TI - Synthesis of some novel imidazolinones as potent anticonvulsant agents. AB - Imidazolinone derivatives of IIa-IIc, IIIa-IIIf and IVa-IVf have been synthesised by the condensation of some known sulpha drugs with 5-oxazolone derivatives, which were prepared by Erlenmeyer condensation of benzoyl glycine with different aldehydes in the presence of sodium acetate and acetic anhydride. The constitution of the products has been supported by elemental analysis and IR, 1H NMR spectral data. The products have been screened for their (a) in vitro growth inhibitory activity against several microorganisms and (b) in vivo anticonvulsant activity. PMID- 14561483 TI - Synthesis and biological activity of 1beta-methyl-2-[5-(2-N-substituted aminoethylcarbamoyl)pyrrolidin-3-ylthio]carbapenem derivatives. AB - The synthesis of a new series of 1beta-methylcarbapenems having the substituted aminoethylcarbamoylpyrrolidine moiety is described. Their in vitro antibacterial activities against both Gram-positive including MRSA and Gram-negative bacteria were tested and the effect of substituent on the pyrrolidine ring was investigated. In particular, the compound 11g having piperazinyl urea moiety showed the most potent antibacterial activity and 11k exhibited excellent anti MRSA. PMID- 14561484 TI - Synthesis and in vitro antibacterial evaluation of N-[5-(5-nitro-2-thienyl)-1,3,4 thiadiazole-2-yl] piperazinyl quinolones. AB - A series of N-[5-(5-nitro-2-thienyl)-1,3,4-thiadiazole-2-yl]piperazinyl quinolones (7a-c) were synthesized and evaluated for in vitro antibacterial activity against some Gram-positive and Gram-negative bacteria. The antibacterial data revealed that compounds 7a-c had strong and better activity against tested Gram-positive organisms than the reference quinolones such as ciprofloxacin, norfloxacin and enoxacin. However, all three compounds were nearly inactive against Gram-negative bacteria. Compound 7a (ciprofloxacin analogue) was the most active compound against Gram-positive bacteria (MIC=0.008-0.015 mug mL(-1)). PMID- 14561485 TI - Effects of CD80 and CD86 on cytokine production in patients with wasp-venom allergy who receive venom immunotherapy. AB - Several studies have provided evidence that activation of antigen-specific T cells requires interactions between CD28 on T cells and its ligands, CD80 and CD86, on antigen-presenting cells (APCs). However, the effects of CD80 and CD86 on cytokine production in patients with Hymenoptera venom allergy who receive venom immunotherapy remain unclear. We examined the effects of CD80 and CD86 on Th1- and Th2-cytokine production before and after venom immunotherapy in patients with wasp-venom allergy. Peripheral blood mononuclear cells (PBMCs) were isolated from patients with wasp-venom allergy before and after three months of venom immunotherapy. CD4+ T cells and monocytes were isolated as APCs from PBMCs and were cocultured with wasp venom in the presence of anti-CD80 or -CD86 blocking antibodies. Interleukin (IL)-4, IL-10, and interferon (IFN)-gamma were measured by enzyme-linked immunosorbent assay. The expression of CD80 and CD86 on CD14+ PBMCs was detected by fluorescence-activated cell-sorter analysis. The expression of CD86, but not that of CD80, on CD14+ PBMCs cocultured with venom increased after three months of venom immunotherapy, but not before venom immunotherapy. Blockade of CD86 reduced IL-10 production after three months of venom immunotherapy. IL-10 production promoted by CD86 costimulation may be involved in the mechanism of venom immunotherapy in patients with venom allergy. PMID- 14561486 TI - Alterations in plasma angiogenic growth factor concentrations after coronary artery bypass graft surgery: relationships with post-operative complications. AB - To determine whether angiogenic growth factor levels are altered during and after cardiac surgery, plasma concentrations of basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF) and transforming growth factor beta1 (TGFbeta1) were measured in 32 patients undergoing coronary artery bypass graft (CABG) surgery with extracorporeal circulation (ECC). EGF levels significantly decreased during ECC and remained low until the 24th post-operative hour with no difference between complicated and uncomplicated patients. TGFbeta1 and bFGF concentrations significantly increased at the end of ECC and after cross-clamp release, and returned to pre-operative values at the 6th post-operative hour suggesting that the source of these elevations are the lungs and heart. After cross-clamp release bFGF levels but not TGFbeta1 ones were higher in patients with respiratory impairments. VEGF values increased significantly at the 6th and 24th post-operative hours. At the 24th post-operative hour plasma VEGF levels were higher in patients with cardiovascular and hematological impairments. In conclusion, these results highlight that the angiogenic network is profoundly altered in patients undergoing cardiopulmonary bypass as previously demonstrated for lipidic, cytokine and haematopoietic growth factor ones and identify an association between specific post-CABG complications and systemic release of bFGF and VEGF. PMID- 14561487 TI - The JNK binding domain of islet-brain 1 inhibits IL-1 induced JNK activity and apoptosis but not the transcription of key proapoptotic or protective genes in insulin-secreting cell lines. AB - The stress-activated protein kinase c-Jun NH2-terminal kinase (JNK) is a central signal for interleukin-1beta (IL-1beta)-induced apoptosis in insulin-producing beta-cells. The cell-permeable peptide inhibitor of JNK (JNKI1), that introduces the JNK binding domain (JBD) of the scaffold protein islet-brain 1 (IB1) inside cells, effectively prevents beta-cell death caused by this cytokine. To define the molecular targets of JNK involved in cytokine-induced beta-cell apoptosis we investigated whether JNKI1 or stable expression of JBD affected the expression of selected pro- and anti-apoptotic genes induced in rat (RIN-5AH-T2B) and mouse (betaTC3) insulinoma cells exposed to IL-1beta. Inhibition of JNK significantly reduced phosphorylation of the specific JNK substrate c-Jun (p<0.05), IL-1beta induced apoptosis (p<0.001), and IL-1beta-mediated c-fos gene expression. However, neither JNKI1 nor JBD did influence IL-1beta-induced NO synthesis or iNOS expression or the transcription of the genes encoding mitochondrial manganese superoxide dismutase (MnSOD), catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase rho (GSTrho), heat shock protein (HSP) 70, IL 1beta-converting enzyme (ICE), caspase-3, apoptosis-inducing factor (AIF), Bcl-2 or Bcl-xL. We suggest that the anti-apoptotic effect of JNK inhibition by JBD is independent of the transcription of major pro- and anti-apoptotic genes, but may be exerted at the translational or posttranslational level. PMID- 14561489 TI - Function of a conserved residue in the amino terminal alpha-helix of four helical bundle cytokines. AB - The Glu residue in the helix A is conserved among many cytokines. Mutation in this residue converts some cytokines to an antagonist. Such an artificial cytokine with an antagonist activity may be useful in a clinical area. In this study we generated a mutant granulocyte colony-stimulating factor (G-CSF) termed G-CSF.E20K in which this residue is substituted to Lys. It is known that G-CSF binds to a homodimeric receptor, while other cytokines which can be converted to antagonists bind to heterodimeric receptors. We showed that G-CSF.E20K does not bind to the receptor at all, and that it fails to stimulate proliferation. Thus, the mutant did not act as an antagonist. We propose that the nature of the receptor, namely whether it is a homodimer or heterodimer, determines the antagonist activity of the mutant. PMID- 14561490 TI - IL-10 induces gene expression in macrophages: partial overlap with IL-5 but not with IL-4 induced genes. AB - The hypothesis that IL-10, in addition to down-regulating pro-inflammatory activities of macrophages, induces an alternative state of macrophage reactivity was tested. We therefore conducted a systematic search for genes induced by IL-10 using the method of suppression subtractive hybridisation. Of an initial 1,300 candidate clones obtained, several screening rounds led to the identification of 51 clones which were reproducibly at least twofold up-regulated in mouse J774 macrophages in response to treatment with IL-10. Of these, 41 genes were homologous to known genes involved in cell metabolism or immunoregulation, five contained novel sequences and another five were homologous to ESTs without known function. One major finding was that about 25% of the IL-10 genes were also found expressed in response to IFNgamma, and several of these also reappeared in IL-4 or IL-5 induced mRNA species. Hence, Th1 and Th2 type cytokines may elicit a common basal activation response in macrophages. The second major finding was that 57% of IL-10 induced genes reappeared in IL-5 induced mRNA but no more than 18% were also found in IL-4 induced mRNA of J774 cells. We conclude that the gene expression response to IL-10 in macrophages is partially different from the response to IL-5 and is substantially different from the response to IL-4, which suggests an unexpected diversity of biological phenotypes induced by different Th2 type cytokines. PMID- 14561488 TI - Hypoxia modulates the effects of transforming growth factor-beta isoforms on matrix-formation by primary human lung fibroblasts. AB - Chronic hypoxia is implicated in lung fibrosis, which is characterized by enhanced deposition of extracellular matrix (ECM) molecules. Transforming growth factor-beta (TGF-beta) plays a key role in fibroblast homeostasis and is involved in disease states characterized by excessive fibrosis, such as pulmonary fibrosis. In this study, we investigated if hypoxia modulates the effects of TGF beta on the expression of gelatinases: matrix metalloproteinase (MMP)-2 and MMP 9, interstitial collagenases: MMP-1 and MMP-13, tissue inhibitors of MMP (TIMP), collagen type I and interleukin-6 (IL-6). Primary human lung fibroblasts, established from tissue biopsies, were cultivated under normoxia or hypoxia in the presence of TGF-beta1, TGF-beta2 or TGF-beta3. Gelatinases were assessed by gelatin zymography and collagenases, TIMP, collagen type I and IL-6 by ELISA. Under normoxia fibroblasts secreted MMP-2, collagenases, TIMP, collagen type I and IL-6. TGF-betas significantly decreased MMP-1 and increased TIMP-1, IL-6 and collagen type I. Hypoxia significantly enhanced MMP-2, and collagenases. Compared to normoxia, the combination of TGF-beta and hypoxia reduced MMP-1, and further amplified the level of TIMP, IL-6, and collagen type I. Thus, in human lung fibroblasts hypoxia significantly increases the TGF-betas-induced secretion of collagen type I and may be associated to the accumulation of ECM observed in lung fibrosis. PMID- 14561492 TI - Looking under every rock: Duchenne muscular dystrophy and traditional Chinese medicine. AB - Traditional Chinese medicine has been advocated to alleviate symptoms in Duchenne muscular dystrophy. To investigate this hypothesis, a pilot study was carried out in Beijing on 10 DMD boys treated with various regimens, including pills, decoctions, massages and acupuncture at various stages of their disease course. Despite the limited scientific impact of such a study, it seems as if the benefit, if any, is minimal. Moreover, some indirect clinical clues such as the cushingoid appearance found in a few patients suggest these drugs may also contain corticosteroids to some extent. PMID- 14561491 TI - Detection of glucocorticoid-like activity in traditional Chinese medicine used for the treatment of Duchenne muscular dystrophy. AB - Anecdotal reports of positive influence of certain traditional Chinese medicines on the progression of neuromuscular diseases in general and Duchenne muscular dystrophy (DMD) in particular has raised interest in patient support groups and clinical experts alike. However, clinical signs of steroid-specific side effects in patients treated with a particular form of Chinese medicine raised the concern that they may contain glucocorticoids, which in turn could also explain the mild beneficial effects seen in some of the patients. We have extracted and fractionated capsules containing pulverized Chinese medicine that had been used for the treatment of DMD patients and analyzed their content for glucocorticoid like activity using promoter-reporter assays. We demonstrate that extracts from this Chinese medicine activate a prototype glucocorticoid-response element, increase the level of utrophin protein in human muscle cells and activate the utrophin promoter A. Based on our bioassays we conclude that this particular Chinese medicine used for the treatment of muscular dystrophy patients contains glucocorticoids as one of its active ingredients. PMID- 14561493 TI - Characterization of Danon disease in a male patient and his affected mother. AB - Danon disease, primary lysosome-associated membrane protein-2 (LAMP-2) deficiency, is histologically characterized by unusual vacuoles bound by membranes with sarcolemmal features in skeletal muscle. We studied skeletal muscle specimens from a male patient with genetically confirmed Danon disease who had two muscle biopsies, at age 20 months and 16 years, and from his mother with cardiomyopathy but without clinically apparent skeletal myopathy. In the patient, the number of vacuoles increased over the 14-year interval between biopsies, suggesting that the number of vacuolated fibers increases with age, and correlates with the development of muscle symptoms. In contrast, in the muscle biopsy from the mother there were no vacuoles even though she had decreased LAMP 2. PMID- 14561494 TI - Involvement of 3Na+/2K+ ATP-ase and Pi-3 kinase in the response of skeletal muscle ATP-sensitive K+ channels to insulin. AB - The modulation of ATP-sensitive K+ channel (K(ATP)) by insulin plays a role in neuromuscular disorders associated to altered K+ homeostasis. However, the mechanisms by which insulin modulates K(ATP) channels are not known. Here, the insulin-dependent 3Na+/2K+ ATP-ase and Pi-3 kinase pathways were explored by using patch-clamp techniques. High and low affinity inhibition of K(ATP) channels by ouabain was observed in the insulin-stimulated and resting fibers, respectively. The 9A5 antibody directed against the alpha1-subunit of the pump inhibited the K(ATP) channel in the resting fibers but fails to inhibit it in the insulin-stimulated fibers. In contrast, the RT2NKATPabr, an alpha2-subunit specific antibody, inhibited the K(ATP) channels in the insulin-stimulated fibers failing to inhibit it in the resting fibers. The insulin-dependent stimulation of K(ATP) channel was prevented by Pi-3 kinase inhibitors Wortmannin and LY294002. In conclusion, insulin stimulating the 3Na+/2K+ ATP-ase activates K(ATP) channels through a membrane-delimited interaction thus controlling the K+ homeostasis. The Pi-3 kinase is the intracellular insulin signal linking the glucose homeostasis to the K(ATP) channel. PMID- 14561495 TI - Identification of novel GDAP1 mutations causing autosomal recessive Charcot-Marie Tooth disease. AB - Mutations in the ganglioside-induced differentiation-associated protein 1 gene cause either autosomal recessive demyelinating Charcot-Marie-Tooth disease type 4A or autosomal recessive axonal Charcot-Marie-Tooth disease with vocal cord paresis. We sequenced the ganglioside-induced differentiation-associated protein 1 gene in 138 patients from 119 unrelated families diagnosed with either demyelinating or axonal autosomal recessive Charcot-Marie-Tooth disease. We detected six distinct mutant alleles in four families, four of which are novel. Electrophysiological studies show severely slowed motor nerve conduction velocities with severely reduced compound muscle action potentials. However, one patient had a normal conduction velocity in the ulnar nerve. Based on the electrophysiological tests, patients with ganglioside-induced differentiation associated protein 1 mutations will therefore be classified as either axonal or demyelinating Charcot-Marie-Tooth disease. The neuropathological aspect shows a divergent pattern; nerve biopsies taken from two siblings at the same age and sharing the same ganglioside-induced differentiation-associated protein 1 gene mutation showed a dissimilar severity stage. PMID- 14561496 TI - Refined mapping of the HMSNR critical gene region--construction of a high-density integrated genetic and physical map. AB - Hereditary motor and sensory neuropathy russe, a form of autosomal recessive Charcot-Marie-Tooth disease, is a rare disorder found in several Roma families from Europe. The gene has been mapped to a 1Mb region on 10q22. Detailed analysis led to the exclusion of 22 candidate genes and the assembly of a high-density genetic map comprising 141 polymorphic markers. Extensive genotyping in an extended sample of affected families resulted in a 10-fold reduction of the critical hereditary motor and sensory neuropathy russe gene region, which is now contained within a single completely sequenced BAC clone. The fact that no sequence variant has been detected in the known genes in the critical region indicates that the hereditary motor and sensory neuropathy russe mutation affects a novel gene that remains to be identified. PMID- 14561497 TI - Sexual differences in onset of disease and response to exercise in a transgenic model of ALS. AB - Transgenic mice that overexpress the mutant human SOD1 gene (hSOD1) serve as an animal model for amyotrophic lateral sclerosis (ALS). Age and sex are recognized as risk factors for ALS, but physical activity remains controversial. Therefore, we investigated the effect of exercise on the phenotype of male and female hSOD1 mice. Onset of disease, progression of disease and survival were measured in low copy and high-copy hSOD1 mice that were randomized to an exercise or sedentary group. We found that onset of disease was different for the two sexes: significantly earlier in male than in female hSOD1 mice. Exercise delayed the onset of disease in female but not in male hSOD1 mice. Also, exercise delayed the total survival time in female high-copy hSOD1 mice. Muscle morphometry and motor neuron counts were similar in all experimental groups at the end of training. Sedentary female hSOD1 mice showed more frequently irregular estrous cycles suggesting a higher estrogen exposure in exercising female mice. These results suggest a possible neuroprotective effect of female sex hormones and support the view that ALS patients should not avoid regular exercise. PMID- 14561498 TI - Comparison of maximal voluntary isometric contraction and hand-held dynamometry in measuring muscle strength of patients with progressive lower motor neuron syndrome. AB - Context. Maximal voluntary isometric contraction, a method quantitatively assessing muscle strength, has proven to be reliable, accurate and sensitive in amyotrophic lateral sclerosis. Hand-held dynamometry is less expensive and more quickly applicable than maximal voluntary isometric contraction. Objective. To investigate if hand-held dynamometry is as reliable and valid as maximal voluntary isometric contraction in measuring muscle strength in patients with an adult-onset, non-hereditary progressive lower motor neuron syndrome. Design. Two testers performed maximal voluntary isometric contraction and hand-held dynamometry measurements in six muscle groups bilaterally in patients with progressive lower motor neuron syndrome to assess reliability and validity of both the methods. Setting. Outpatient units of an academic medical center. Patients. A consecutive sample of 19 patients with non-hereditary progressive lower motor neuron syndrome (median disease duration 32.5 months, range 10-84) was tested. Outcome measures. Comparison between maximal voluntary strength contractions as measured by hand-held dynamometry and maximal voluntary isometric contraction. Results. Low intra- and interrater variation in all muscle groups were found, intraclass correlation coefficients vary between 0.86 and 0.99 for both methods. Both methods correlated well in all muscle groups with Pearson's correlation coefficients ranged between 0.78 and 0.98. Scatter plots indicated a trend to under-estimate muscle strength above 250 N by hand-held dynamometry as compared with maximal voluntary isometric contraction. Conclusions. For longitudinal evaluation of muscle strength in patients with progressive lower motor neuron syndrome (i.e. between 0 and 250 N), muscle strength can be accurate quantified with both hand-held dynamometry and maximal voluntary isometric contraction. Hand-held dynamometry has the advantage of being cheap and quickly applicable. However, our results indicate that hand-held dynamometry is less sensitive than maximal voluntary isometric contraction in detecting subnormal muscle strength in strong muscle groups (i.e. >250 N), due to limited strength of the tester. PMID- 14561499 TI - Paediatric mononeuritis multiplex: a report of three cases and review of the literature. AB - Mononeuritis multiplex results from disease processes causing multifocal injury to the peripheral nerves, with or without involvement of one or more cranial nerves. Most reported cases of paediatric mononeuritis multiplex have been related to autoimmune disorders. We present clinical and neurophysiologic findings in three cases of multiple mononeuropathy occurring in adolescence, with onset at ages 16, 17 and 13 years. Two of these cases were related to systemic vasculitis, one possibly representing a paediatric case of non-systemic vasculitis. Sural nerve biopsy confirmed the diagnosis of vasculitis in one patient with systemic lupus erythematosus, but was non-diagnostic in another case. Mononeuritis multiplex is an extremely uncommon form of acquired peripheral neuropathy in childhood. We discuss the differential diagnosis and review previous descriptions of this rare condition. PMID- 14561500 TI - 116th ENMC international workshop: the treatment of mitochondrial disorders, 14th 16th March 2003, Naarden, The Netherlands. PMID- 14561505 TI - Global phytochemistry: research in Japan. PMID- 14561506 TI - Stevioside. AB - Stevioside is a natural sweetener extracted from leaves of Stevia rebaudiana (Bertoni) Bertoni. The literature about Stevia, the occurrence of its sweeteners, their biosynthetic pathway and toxicological aspects are discussed. Injection experiments or perfusion experiments of organs are considered as not relevant for the use of Stevia or stevioside as food, and therefore these studies are not included in this review. The metabolism of stevioside is discussed in relation with the possible formation of steviol. Different mutagenicity studies as well as studies on carcinogenicity are discussed. Acute and subacute toxicity studies revealed a very low toxicity of Stevia and stevioside. Fertility and teratogenicity studies are discussed as well as the effects on the bio availability of other nutrients in the diet. The conclusion is that Stevia and stevioside are safe when used as a sweetener. It is suited for both diabetics, and PKU patients, as well as for obese persons intending to lose weight by avoiding sugar supplements in the diet. No allergic reactions to it seem to exist. PMID- 14561507 TI - Analysis and biological activities of anthocyanins. AB - Anthocyanins are naturally occurring compounds that impart color to fruits, vegetables, and plants. They are probably the most important group of visible plant pigments besides chlorophyll. Apart from imparting color to plants, anthocyanins also have an array of health-promoting benefits, as they can protect against a variety of oxidants through a various number of mechanisms. However, anthocyanins have received less attention than other flavonoids, despite this. This article reviews their biological functions and pre-clinical studies, as well as the most recent analytical techniques concerning anthocyanin isolation and identification. PMID- 14561508 TI - Beta-amylase in germinating millet seeds. AB - Beta-amylase (EC 3.2.1.2) was isolated from germinating millet (Panicum miliaceum L.) seeds by a procedure that included ammonium sulfate fractionation, chromatography on DEAE-cellulofine and CM-cellulofine, and preparative isoelectric focusing. The enzyme was homogeneous by SDS-PAGE. The M(r) of the enzyme was estimated to be 58,000 based on its mobility on SDS-PAGE and gel filtration with TSKgel G4000SW(XL), which showed that it is composed of a single unit. The isoelectric point of the enzyme was 4.62. The enzyme hydrolyzed malto oligosaccharides more readily as their degree of polymerization increased, this being strongest for malto-oligosaccharides larger than 13 glucose residues and very weakly for maltotriose. Amylose, amylopectin and soluble starch were the most suitable substrates for the enzyme. While the enzyme showed some activity against native starch by itself, starch digestion was accelerated 2.5-fold using alpha-amylase, pullulanase and alpha-glucosidase. This enzyme appears to be very important for the germination of millet seeds. PMID- 14561509 TI - Effect of salt stress on the metabolism of ethanolamine and choline in leaves of the betaine-producing mangrove species Avicennia marina. AB - Glycinebetaine synthesis from [methyl-14C]choline and [1,2-14C]ethanolamine in leaf disks of Avicennia marina, was increased by salt stress (250 and 500 mM NaCl). After 18 h incubation with [methyl-14C]choline, phosphocholine and CO(2) were found to be heavily labelled. Phosphocholine contained 39% of the total radioactivity taken up by non-salinised (control) leaf disks and 15% of the total for salinised leaf disks stressed with 500 mM NaCl. Eighteen and 49% of the radioactivity absorbed by control and salinised disks, respectively, were released as CO(2). Metabolic studies of [1,2-14C]ethanolamine revealed that the radioactivity taken up by the leaf disks was recovered as the following compounds after 18 h: phosphorylated compounds (mainly phosphoethanolamine, phosphodimethylethanolamine and phosphocholine) (40-50%); choline (1-2%); glycinebetaine (3-5%); lipids (20-28%); CO(2) (6-10%). Unlike glycinebetaine, incorporation into phosphorylated compounds and lipids were reduced by salt stress. Incorporation of [methyl-14C]S-adenosyl-L-methionine (SAM) into choline, phosphocholine and glycinebetaine in leaf disks was stimulated by salt stress. In vitro activities of adenosine kinase and adenosine nucleosidase, which are implicated in stimulating the SAM regeneration cycle, increased after the leaf disks were incubated with 250 and 500 mM NaCl for 18 h. Changes in metabolism involving choline and glycinebetaine due to salt stress are discussed. PMID- 14561510 TI - Phytoalexins from Thlaspi arvense, a wild crucifer resistant to virulent Leptosphaeria maculans: structures, syntheses and antifungal activity. AB - Phytoalexins are inducible chemical defenses produced by plants in response to diverse forms of stress, including microbial attack. Our search for phytoalexins from cruciferous plants resistant to economically important fungal diseases led us to examine stinkweed or pennycress (Thlaspi arvense), a potential source of disease resistance to blackleg. We have investigated phytoalexin production in leaves of T. arvense under abiotic (copper chloride) and biotic elicitation by Leptosphaeria maculans (Desm.) Ces. et de Not. [asexual stage Phoma lingam (Tode ex Fr.) Desm.], and report here two phytoalexins, wasalexin A and arvelexin (4 methoxyindolyl-3-acetonitrile), their syntheses and antifungal activity against isolates of P. lingam/L. maculans, as well as the isolation of isovitexin, a constitutive glycosyl flavonoid of stinkweed, having antioxidant properties but devoid of antifungal activity. PMID- 14561511 TI - Transformation of the host-selective toxin destruxin B by wild crucifers: probing a detoxification pathway. AB - The destruxin B detoxification pathway present in Sinapis alba is also present in three unrelated species, Camelina sativa, Capsella bursa-pastoris, and Eruca sativa, suggesting a conservation of this pathway across crucifers. The chemical structure of a destruxin B metabolite, (6'-O-malonyl)hydroxydestruxin B beta-D glucopyranoside, was also establised. Considering that Camelina sativa and Capsella bursa-pastoris detoxify destruxin B and produce the phytoalexins camalexins, these wild crucifers appear to represent unique and perhaps useful sources of blackleg resistance in strategic plant breeding. PMID- 14561512 TI - Phenylpropanoid amides of serotonin accumulate in witches' broom diseased bamboo. AB - Aciculosporium take (Ascomycota; Clavicipitaceae), causes the witches' broom disease in bamboo, particularly Phyllostachys bambusoides. Since it was observed that endogenous indole-3-acetic acid is reduced in the twigs of the diseased bamboo, the symptoms (bushy appearance) may be induced by reduction in auxin levels. Furthermore, two indolic compounds accumulated in diseased twigs, these being identified as N-p-coumaroylserotonin and N-feruloylserotonin by LC-MS, 1H NMR and 13C NMR spectroscopic analyses. N-p-Coumaroylserotonin possesses antifungal activity against A. take. PMID- 14561513 TI - Occurrence of the allene oxide cyclase in different organs and tissues of Arabidopsis thaliana. AB - Occurrence of an essential enzyme in jasmonate (JA) biosynthesis, the allene oxide cyclase, (AOC) was analyzed in different developmental stages and various organs of Arabidopsis thaliana plants by immuno blot analysis and immunocytological approaches. Levels of AOC and of the two preceding enzymes in JA biosynthesis increased during seedling development accompanied by increased levels of JA and 12-oxophytodienoic acid levels after 4 and 8 weeks. Most tissues including all vascular bundles and that of flower buds contain AOC protein. Flowers shortly before opening, however, contain AOC protein preferentially in ovules, stigma cells and vascular bundles, whereas in anthers and pollen AOC could not be detected. The putative roles of AOC and JA in development are discussed. PMID- 14561514 TI - Prenylated xanthones with NGF-potentiating activity from Garcinia xanthochymus. AB - Two prenylated xanthones, 1,4,5,6-tetrahydroxy-7,8-di(3-methylbut-2-enyl)xanthone (1) and 1,2,6-trihydroxy-5-methoxy-7-(3-methylbut-2-enyl)xanthone (2), were isolated from the wood of Garcinia xanthochymus along with a known xanthone, 12b hydroxy-des-D-garcigerrin A (3). Their structures were elucidated by spectroscopic analysis. Compound 1 (10 muM), 2 (10-30 muM) and 3 (10 muM) showed a markedly enchancing activity of nerve growth factor (NGF)-mediated neurite outgrowth on PC12D cells. PMID- 14561516 TI - Two lignan dimers from bamboo stems (Phyllostachys edulis). AB - Phyllostadimers A and B, two bis-lignans in which the two lignan units are directly connected by a C-C bond, were isolated from stems of bamboo, Phyllostachys edulis. Their structures were determined on the basis of spectral evidence. In addition, 14 known compounds were also obtained throughout the investigation. Phyllostadimer A significantly inhibited liposomal lipid peroxidation. PMID- 14561515 TI - Phenolic derivatives from Wigandia urens with weak activity against the chemokine receptor CCR5. AB - Three compounds, 2,3-dihydroxy-4-methoxy-6,6,9-trimethyl-6H-dibenzo[b,d]pyran (1), 8-methoxy-2-methyl-2-(4-methyl-3-pentenyl)-2H-1-benzopyran-6-ol (2) and 4 methoxy-3-(3-methyl-2-butenyl)-benzoic acid (3), have been isolated from Wigandia urens. The structures of compounds 1, 2 and 3 were determined from spectroscopic data and showed activity in a CCR5 assay with IC(50) values of 33, 46 and 26 muM respectively. PMID- 14561517 TI - Isolation and anti-oomycete activity of nyasol from Anemarrhena asphodeloides rhizomes. AB - The methanol extract of Anemarrhena asphodeloides rhizomes exhibited strong antifungal activity against the plant pathogenic fungi Magnaphothe grisea, Rhizoctonia solani, and the plant pathogenic oomycete Phytophthora capsici. The antifungal substance isolated from the rhizomes of A. asphodeloides was identified to be nyasol, (Z)-1,3-bis(4-hydroxyphenyl)-1,4-pentadiene by NMR and mass spectral analysis. Nyasol effectively inhibited the mycelial growth of Colletotrichum orbiculare, P. capsici, Pythium ultimum, R. solani, and Cladosporium cucumerinum in a range of 1-50 mug/ml, but did not affect the growth of bacteria and yeast. In a greenhouse test, treatment with the antifungal compound nyasol was significantly effective in suppressing the Phytophthora blight on pepper plants. PMID- 14561519 TI - Profiling phenolic metabolites in transgenic alfalfa modified in lignin biosynthesis. AB - Soluble phenolics, wall-bound phenolics and soluble and core lignin were analyzed in transgenic alfalfa with genetically down-regulated O-methyltransferase genes involved in lignin biosynthesis. High performance liquid chromatography and principal component analysis were used to distinguish metabolic phenotypes of different transgenic alfalfa genotypes growing under standard greenhouse conditions. Principal component analysis of HPLC chromatograms did not resolve differences in leaf metabolite profiles between wild-type and transgenic plants of the same genetic background, although stem phenolic profiles were clearly different between wild-type and transgenic plants. However, the analytical methods clearly differentiated two non-transgenic alfalfa cultivars based on either leaf or stem profiles. Metabolic profiling provides a useful approach to monitoring the broader biochemical phenotypes of transgenic plants with altered expression of lignin pathway enzymes. PMID- 14561518 TI - Polar constituents of celery seed. AB - From the water-soluble portion of the methanol extract of celery seed (fruit of Apium graveolens L.) five sesquiterpenoid glucosides (celerioside A-E) and three phthalide glycosides (celephtalide A-C) were isolated together with six aromatic compound glucosides, two norcarotenoid glucosides and a lignan glucoside. Their structures were determined by spectral investigations. PMID- 14561520 TI - Changes in lignin content of leaf litters during mulching. AB - Alkaline nitrobenzene oxidation, ozonation and methoxyl content determinations were applied to decomposing leaf litter of Ginkgo biloba L., Cinnamomum camphora sieb., Zelkova serrata Makino and Firmiana simplex W. F. Wight, respectively, during mulching to investigate the properties and estimate changes in lignin composition and content. Since the Klason lignin residue originated from components highly resistant to degradation by acid, the methoxyl content of Klason residue was used to estimate the lignin content of leaf litter. Quantitative analysis of presumed lignin-derived fragments, by use of alkaline nitrobenzene oxidation and ozonation methods, suggested that the estimated lignin content approximates that of the real lignin content of leaves, which is greatly overestimated by the Klason procedure. The estimated lignin contents ranged from 3.9 to 10.0% while the Klason lignan residue varied from 37.1 to 46.7% in un mulched leaf litter. The absolute amounts of the measured lignin somewhat decreased during mulching, while the structure of lignin remaining in leaf litters after mulching was considered not to be very different from its original structure. PMID- 14561523 TI - Lowered serum n-3 polyunsaturated fatty acid (PUFA) levels predict the occurrence of postpartum depression: further evidence that lowered n-PUFAs are related to major depression. AB - Several studies have shown that major depression is accompanied by alterations in serum fatty acid composition, e.g. reduced n-3 fatty acids and an increased 20:4n 6/20:5n-3 ratio in serum. Moreover, pregnancy leads to depletion of maternal serum 22:6n-3 and after delivery maternal serum 22:6n-3 steadily declines further. Therefore, the aim of the present study was to investigate whether the postpartum fatty acid profile of maternal serum phospholipids (PL) and cholesteryl esters (CE) differs in women who develop postpartum depression compared to controls. We compared the fatty acid composition shortly after delivery of 10 women who developed postpartum depression and 38 women who did not. After delivery, 22:6n-3 and the sum of the n-3 fatty acids in PL and CE was significantly lower in the group of mothers who developed a postpartum depression. The ratio of Sigman-6/Sigman-3 fatty acids in PL was, postpartum, significantly higher in the depressed group as compared to the controls. The abnormalities in fatty acid status previously observed in major depression are now also confirmed in postpartum depression. These results indicate that pregnant women who are at risk to develop postpartum depression may benefit from a prophylactic treatment with n-3 PUFAs, such as a combination of 20:5n-3 and 22:6n 3. PMID- 14561522 TI - "Broad spectrum" antidepressants: is more better for the treatment of depression? AB - The majority of antidepressants in current use selectively inhibit the reuptake of serotonin and/or norepinephrine. "Broad spectrum" antidepressants are compounds that inhibit the reuptake of norepinephrine, serotonin and dopamine, the three biogenic amines most closely linked to depression. The pharmacological profile of one such compound has recently been described (European Journal of Pharmacology, 461 (2003) 99). DOV 21,947, an azabicyclo[3.1.0]hexane, potently inhibits norepinephrine, serotonin and dopamine reuptake by the corresponding human transporter proteins. DOV 21,947 is orally active in the forced swim and tail suspension tests, preclinical procedures that are highly predictive of antidepressant action in patients. A closely related compound, DOV 216,303 is safe and well-tolerated in Phase I studies. The plasma concentrations of DOV 216,303 following both single and multiple doses appear sufficient to inhibit norepinephrine, serotonin, and dopamine reuptake. Based on the pivotal role proposed for dopamine in depression, it has been hypothesized that a broad spectrum antidepressant will produce a more rapid onset and/or higher efficacy than agents inhibiting the reuptake of serotonin and/or norepinephrine. PMID- 14561524 TI - Tissue microarray study for classification of breast tumors. AB - Clinical and pathological heterogeneity of breast cancer hinders selection of appropriate treatment for individual cases. Molecular profiling at gene or protein levels may elucidate the biological variance of tumors and provide a new classification system that correlates better with biological, clinical and prognostic parameters. We studied the immunohistochemical profile of a panel of seven important biomarkers using tumor tissue arrays. The tumor samples were then classified with a monothetic (binary variables) clustering algorithm. Two distinct groups of tumors are characterized by the estrogen receptor (ER) status and tumor grade (p = 0.0026). Four biomarkers, c-erbB2, Cox-2, p53 and VEGF, were significantly overexpressed in tumors with the ER-negative (ER-) phenotype. Eight subsets of tumors were further identified according to the expression status of VEGF, c-erbB2 and p53. The malignant potential of the ER-/VEGF+ subgroup was associated with the strong correlations of Cox-2 and c-erbB2 with VEGF. Our results indicate that this molecular classification system, based on the statistical analysis of immunohistochemical profiling, is a useful approach for tumor grouping. Some of these subgroups have a relative genetic homogeneity that may allow further study of specific genetically-controlled metabolic pathways. This approach may hold great promise in rationalizing the application of different therapeutic strategies for different subgroups of breast tumors. PMID- 14561525 TI - Metabolism of medroxyprogesterone acetate (MPA) via CYP enzymes in vitro and effect of MPA on bleeding time in female rats in dependence on CYP activity in vivo. AB - Medroxyprogesterone acetate (MPA) is a drug commonly used in endocrine therapy for advanced breast cancer, although it is known to cause thrombosis as a serious side effect. Recently, we found that cytochrome P450 3A4 (CYP3A4) mainly catalyzed the metabolism of MPA via CYP in human liver microsomes. However, the metabolic products of MPA in humans and rats have not been elucidated. In addition, it is not clear whether thrombosis could be induced by MPA itself or by its metabolites. In this study, we determined the overall metabolism of MPA as the disappearance of the parent drug from an incubation mixture, and identified the enzymes catalyzing the metabolism of MPA via CYP in rats. Moreover, the effects of CYP-modulators on MPA-induced hypercoagulation in vivo were examined. Intrinsic clearance of MPA in rat liver microsomes was increased by treatment with CYP3A-inducers. The intrinsic clearance of MPA in liver microsomes of rats treated with various CYP-inducers showed a significant correlation with CYP3A activity, but not CYP1A activity, CYP2B activity or CYP2C contents. Among the eight recombinant rat CYPs studied, CYP3A1, CYP3A2 and CYP2A2 catalyzed the metabolism of MPA. However, since CYP3A2 and CYP2A2 are male-specific isoforms, CYP3A1 appears to be mainly involved in the metabolism of MPA in liver microsomes of female rats. In an in vivo study, pretreatment of female rats with SKF525A, an inhibitor of CYPs including CYP3A1, significantly (p < 0.05) enhanced MPA-induced hypercoagulation, whereas pretreatment with phenobarbital, an inducer of CYPs including CYP3A1, reduced it. These findings suggest that CYP-catalyzed metabolism of MPA is mainly catalyzed by CYP3A1 and that MPA-induced hypercoagulation is predominantly caused by MPA itself in female rats. PMID- 14561526 TI - Calmodulin gene expression in the neural retina of the adult rat. AB - Calmodulin (CaM) mRNAs are expressed with low abundancy in the adult rat neural retina. However, when digoxigenin (DIG)-labeled cRNA probes specific for each CaM mRNA population were hybridized at slightly alkaline pH (pH 8.0), the widespread distribution of CaM mRNA-expressing cells was revealed, with similar abundance for all three CaM genes. The CaM genes displayed a uniquely similar, layer specific expression throughout the retina, and no significant differences were found in the distribution patterns of the CaM mRNA populations or the labeled cell types. The strongest signal for all CaM mRNAs was demonstrated in the ganglion cell layer and the inner nuclear layer, where the highest signal intensity was found within the inner sublamina. Similarly intermediate signal intensities for all CaM genes were detected in the inner and outer plexiform layers, within the vicinity of the outer limiting membrane and in the retinal pigment epithelium. A very low specific signal was characteristic in the outer nuclear layer and the photoreceptor inner segment layer, while no specific hybridization signal was observed in the photoreceptor outer segment layer. In summary, all CaM genes exhibited a similar and a characteristically layer specific expression pattern in the adult rat retina. PMID- 14561527 TI - Polyozellus multiplex, a Korean wild mushroom, as a potent chemopreventive agent against stomach cancer. AB - Polyozellus multiplex, a Korean wild mushroom, was extracted using methanol, and the extract was further fractionated with water and ethylacetate. Assay of each fraction with MTT revealed significant tumoristatic effects of the water fraction of Polyozellus multiplex against human gastric and other cancer cells but not normal human lymphocytes. Modifying effects of the water fraction on glandular stomach mucosa were investigated in male Wistar rats treated with N-methyl-N' nitro-N-nitrosoguanidine (MNNG). The dietary 0.5% or 1% water fraction of Polyozellus multiplex significantly increased glutathione S-transferase (GST) and superoxide dismutase (SOD) activities, and showed a tendency for increase in glutathione (GSH) levels, compared to the MNNG alone group. It also caused a significant reduction in proliferating cell nuclear antigen (PCNA)-labeling index of the glandular stomach epithelium, along with increase in p53 tumor suppressor gene expression. These results suggest that Polyozellus multiplex is a candidate for chemoprevention against gastric cancer. PMID- 14561528 TI - Effect of losartan on the blood-brain barrier permeability in diabetic hypertensive rats. AB - Our previous publication has stressed the benefits of losartan, an angiotensin II receptor blocker, on the permeability of blood-brain barrier (BBB) and blood pressure during L-NAME-induced hypertension. This study reports the impacts of anti-hypertensive treatment by losartan on the brain endothelial barrier function and the arterial blood pressure, during acute hypertension episode, in experimentally diabetic hypertensive rats. Systolic blood pressure measurements were taken with tail cuff method before and during administration of L-NAME (0.5 mg/ml). We induced diabetes by using alloxan (50 mg/kg, i.p). Losartan (3 mg/kg, i.v) was given to rats following the L-NAME treatment. Acute hypertensive vascular injury was induced by epinephrine (40 microg/kg). The BBB disruption was quantified according to the extravasation of the Evans blue (EB) dye. L-NAME induced a significant increase in arterial blood pressure on day 14 in normoglycemic and hyperglycemic rats (p < 0.05). Losartan significantly reduced the increased blood pressure in hypertensive and diabetic hypertensive rats (p < 0.01). Epinephrine-induced acute hypertension in diabetic hypertensive rats increased the content of EB dye dramatically in cerebellum and diencephalon (p < 0.01) and slightly in both cerebral cortex (p < 0.05). Losartan treatment reduced the increased BBB permeability to EB dye in the brain regions of diabetic hypertensive rats treated with epinephrine (p < 0.05). This study indicates that, in diabetic hypertensive rats, epinephrine administration leads to an increase in microvascular-EB-albumin efflux to brain, however losartan treatment significantly attenuates this protein's transport to brain tissue. PMID- 14561530 TI - Anxiolytic-like effect of succinic acid in mice. AB - The putative anxiolytic activity of succinic acid was examined in male mice by using a number of experimental paradigms of anxiety and compared with that of the known anxiolytic compound diazepam. Use of the elevated plus-maze test revealed that diazepam (1.0, 2.0 and 4.0 mg/kg, PO) or succinic acid (3.0 or 6.0 mg/kg, PO) increased the percentage of entries into open arms and of time spent on open arms. In novel food consumption test, succinic acid (3.0, 6.0, and 12.0 mg/kg, IP) caused significant increases in food intake during 5 min when compared with the vehicle. In the stress-induced hyperthermia test, 40 min after drug administration rectal temperature was measured, succinic acid at dose of 1.5 mg/kg, inhibited stress-induced hyperthermia. Thus, these findings indicated that, in contrast with diazepam, succinic acid exhibits anxiolytic-like effect. PMID- 14561529 TI - Tochu (Eucommia ulmoides) leaf extract prevents ammonia and vitamin C deficiency induced gastric mucosal injury. AB - The ingestion of dietary antioxidants, including vitamin C (VC), is suggested to play an important role in the prevention of gastric cancer associated with Helicobacter pylori (HP) infection. Recently, water extracts of Tochu (Du-zhong, Eucommia ulmoidea OLIVER) leaves (WETL) have been reported to have potent antioxidant and antimutagenic effects. The present study investigated the effect(s) of VC and WETL on gastric mucosal injury induced by ammonia and a VC deficient diet. Guinea pigs fed the water containing ammonia and/or a VC deficient diet were simultaneously treated with WETL or VC. Intramucosal levels of thiobarubiturate reactive substances (TBARS), an index of lipid peroxidation, increased significantly in animals fed ammoniated water and VC-deficient diets. This was accompanied by accelerated cell proliferation and increases in immunohistochemical staining indices for oxidative stress-induced DNA adducts and strand breaks (e.g., BrdU-uptake, 8-OhdG, ssDNA and the TUNEL reaction). The administration of either WETL or VC to the ammoniated water and VC-deficient diets ameliorated the increases in intramucosal TBARS levels and labeling indices of BrdU, 8-OHdG, ssDNA and TUNEL, i.e., the levels were similar to those measured in the normal-fed control animals. These data suggest that insufficient VC ingestion may be an important risk factor for gastric cancer development in patients with HP infections. Furthermore, our results suggest that WETL or some constituent may contribute to the prevention of oxidative gastric injury that precedes carcinogenesis. PMID- 14561531 TI - The MEF2 site is necessary for induction of the myosin light chain 2 slow promoter in overloaded regenerating plantaris muscle. AB - Functional overload (OV) of the rat plantaris muscle results in a fast to slow change in muscle phenotype with induction of the slow contractile protein genes including myosin light chain 2 slow (MLC2s). To identify potential cis-acting DNA sites regulating MLC2s following ablation, plasmid constructs were transfected in vivo into regenerating overloaded plantaris muscles. Activity of the 270bp promoter (-270MLC2s) was increased in OV muscles at 28 days. Mutation of the MEF2 site (-270MEF2) knocked out the overload-induced activity of the promoter. Mutation of the Ebox (-270Ebox) resulted in an earlier induction with OV and mutation of the CACC site (-270CACC) resulted in increased activity in the CON PLN with OV induction detected by 21 days. These results demonstrate that the 270MLC2s promoter contains the elements necessary for expression of MLC2s in regenerating OV PLN. More importantly, mutation analysis of -270MLC2s promoter demonstrates that mechanical loading induced expression shares some common molecular mechanisms with slow nerve dependent model regulation. In these two models of physiological induction of MLC2s, the CACC site acts as a repressor region (on/off switch) and the MEF2 site acts to modulate quantitative expression. PMID- 14561533 TI - The importance of the prolonged postnatal period in determination of heart atrial muscarinic receptor and beta-adrenoceptor densities. AB - Our previous work indicated that hyperstimulation of muscarinic receptors brought about profound changes not only in the density of the muscarinic receptors, but also of the beta-adrenoceptors both in vivo and in vitro. Now we try to identify the changes in receptor densities when the neuronal input from the autonomic nerves is disrupted, i.e. when neonatal rats were injected with 6-hydroxydopamine (6-OHDA; able to destroy sympathetic nerves). Although the interruption of neuronal input was successful as indicated by norepinephrine uptake, there was no change in receptor densities in the rat heart atria when 6-OHDA was applied in the first postnatal week only. When we repeated the 6-OHDA treatment consequently (on 14th, 21st and 28th postnatal day) there were clear increases of both receptor types (muscarinic receptors rise to 139% of control, beta-adrenoceptors to 134% of control). The atrial muscarinic receptor number was increased (to 146% of control) also when we combined the first week 6-OHDA application with another factor (when the pups abide with mother till the 56th day). The simple non weaning of animals has no effect on the receptor densities. We can therefore conclude that the first week application of 6-OHDA was insufficient in changing the receptor number and that only repeated 6-OHDA administration was able to change them. Similarly, combination with other factor: non-weaning led to the muscarinic receptors increase. Our results can indicate the importance of the prolonged postnatal period for the heart muscarinic and beta-adrenergic receptor number determination. PMID- 14561532 TI - Mechanism for generation of hydrogen peroxide and change of mitochondrial membrane potential during rotenone-induced apoptosis. AB - Rotenone, an inhibitor of NADH dehydrogenase complex, is a naturally occurring insecticide, which is capable of inducing apoptosis. Rotenone-induced apoptosis is considered to contribute to its anticancer effect and the etiology of Parkinson's disease (PD). We demonstrated that rotenone induced internucleosomal DNA fragmentation, DNA ladder formation, in human cultured cells, HL-60 (promyelocytic leukemia) and BJAB cells (B-cell lymphoma). Flow cytometry showed that rotenone induced H2O2 generation, followed by significant changes in the mitochondrial membrane potential (DeltaPsim). Caspase-3 activity increased in HL 60 cells in a time-dependent manner. These apoptotic events were delayed in HP100 cells, an H2O2-resistant clone of HL-60, confirming the involvement of H2O2 in apoptosis. Expression of anti-apoptotic protein, Bcl-2, in BJAB cells drastically inhibited DeltaPsim change and DNA ladder formation but not H2O2 generation, confirming the participation of mitochondrial dysfunction in apoptosis. NAD(P)H oxidase inhibitors prevented H2O2 generation and DNA ladder formation. These results suggest that rotenone induces O2(-)-derived H2O2 generation through inhibition of NADH dehydrogenase complex and/or activation of NAD(P)H oxidase, and H2O2 generation causes the disruption of mitochondrial membrane in rotenone induced apoptosis. PMID- 14561534 TI - Programmed cell death in Xenopus laevis spinal cord, tail and other tissues, prior to, and during, metamorphosis. AB - Programmed cell death is necessary for the shaping and remodelling of nervous and non-nervous tissues during development. Amphibia, whose body undergoes profound modifications during metamorphosis, are particularly useful models for studying the relationship between cell death in muscles and other non-nervous tissues on the one hand, and in the nervous system connected with these tissues on the other hand. We checked the occurrence of apoptotic cells (identified by TUNEL labelling) in different organs and regions from hatching (stages 35-36) to climax (stages 63-64) in the African Clawed Frog Xenopus laevis. Some organs (e.g., skin and digestive tract) contained apoptotic cells during the entire period studied. In transitory organs (cement gland and gills), a single wave of cell death occurred during the regression of these tissues. In order to compare the timing of cell death in the spinal cord with that of tail regression, we counted the number of TUNEL-positive cells in spinal cord sections taken from animals between stages 54 and 64. Three-dimensional reconstructions using confocal microscopy of vibratome slices immunostained for the detection of c-Jun-like protein accumulated in the cytoplasm of apoptotic cells showed numerous cells at various degrees of degeneration. Many of these cells still presented the morphological characteristics of neurones. The peak of apoptosis was found at stage 58, preceding tail regression. This suggests that neural cell death is not a consequence but rather an element upstream in the chain of events leading to tail degeneration. PMID- 14561535 TI - The war on words: sensible compromise or slow suicide? PMID- 14561536 TI - Comparative study of the effects of two once-a-month injectable contraceptives (Cyclofem and Mesigyna) and one oral contraceptive (Ortho-Novum 1/35) on coagulation and fibrinolysis. AB - A randomized controlled multicenter study was undertaken to monitor the effects on hemostasis of two once-a-month injectable contraceptive preparations, Mesigyna (50 mg norethisterone enanthate and 5 mg estradiol valerate) and Cyclofem (25 mg medroxyprogesterone acetate and 5 mg estradiol cypionate) in comparison with a well-known oral contraceptive (OC) Ortho-Novum 1/35 (norethisterone 1 mg and ethinyl estradiol 35 microg). A total of 378 volunteers from four centers (Bangkok, Hangzhou, Santiago and Singapore) were monitored. Blood sampling took place in one pretreatment cycle, the third and ninth injection intervals and one posttreatment cycle. In each of the three treatment groups, a rise in hemoglobin, and increases in platelet count and in prothrombin time were observed. With treatment there was a significant increase in activated partial thromboplastin time among Mesigyna users, no change among Cyclofem users and a significant decrease among OC users. OC use led to increases in plasma levels of fibrinogen, factor VII, factor X, plasminogen, protein C and decreases in plasma levels of t PAI and antithrombin. Use of combined injectables induced no change (Cyclofem) or decreases (Mesigyna) in plasma levels of fibrinogen, factor VII, factor X and antithrombin. Use of both combined injectables led to decreases in protein C, slight decreases in plasminogen and increases in plasminogen and fibrinogen. Overall, the injectable preparations may be more beneficial than the oral preparation in not enhancing a hypercoagulable state because of the reduced synthesis of fibrinogen, factors VII and X; however, decreases in antithrombin and protein C, which are potent coagulation inhibitors, may raise some concern. Whether these changes can lead to modifications in the risk of arterial or venous disease can only be ascertained by conducting epidemiological studies. PMID- 14561537 TI - Longitudinal changes in bone density in relation to oral contraceptive use. AB - The primary aim of this 36-month prospective cohort study was to evaluate the association between use of oral contraceptives (OCs) and bone mineral density in reproductive-age women. The 36-month bone density (g/cm2) at the spine, hip and whole body and percent change from baseline (measured at 6-month intervals) were evaluated among 245 women 18-39 years of age; 89 were using OCs (median duration: 3.7 years at study entry) and 156 were not using any hormonal contraception. Before and after adjustment for covariates (baseline bone density, age, race, ever pregnant, exercise, body mass and calcium intake), women using OCs did not differ significantly from comparison women in percent change in bone density over 36 months or in absolute bone density at 36 months. All p-values for between group differences were >0.55. In conclusion, within the limitations of this study, OCs did not appear to impact bone density over time in this cohort of reproductive-age women. PMID- 14561538 TI - Effect of a low-dose oral contraceptive containing 20 microg ethinylestradiol and 150 microg desogestrel on dysmenorrhea. AB - This observational study evaluated the effect of a low-dose oral contraceptive (OC), containing 20 mug ethinylestradiol and 150 mug desogestrel (20EE/150DSG) on dysmenorrhea. Of the 346 women enrolled, 301 (87%) completed the three-cycle study. All participants (mean age: 22.6 years) wished to use oral contraception and had dysmenorrhea for at least 3 months. Dysmenorrhea was assessed using both the adapted verbal multidimensional scoring system (VMSS-A) and a visual analogue scale (VAS) to rate pain intensity. General dysmenorrhea symptoms and the use of analgesics were also assessed. The mean total combined scores on the VMSS-A scale decreased from 5.4 to 1.4 (p < 0.001), during the three cycles of OC use. The mean VAS pain-intensity score decreased from 6.8 to 1.8 (p < 0.001). During treatment there were substantial improvements in most individual parameters of dysmenorrhea. The percentage of women with dysmenorrhea lasting > or =2 days decreased from 81% at baseline to 14% at cycle 3. The percentage of women reporting generalized dysmenorrhea symptoms (e.g., lumbago, asthenia) decreased from 79% to 21% and the percentage needing analgesics dropped from 100% to 41%. The percentage of women for whom dysmenorrhea interfered with daily activities decreased from 73% to 10%. The results indicate that 20EE/150DSG can statistically and clinically significantly reduce the incidence and severity of dysmenorrhea, and its impact on daily activities. PMID- 14561539 TI - Effect of Implanon and Norplant subdermal contraceptive implants on serum lipids- a randomized comparative study. AB - The objective of this study was to assess the possible differences in effects of Implanon (etonogestrel 68 mg, single-rod) and Norplant (levonorgestrel 36 mg, six capsules) implants on serum lipids over 2 years of use. In this 2-year open randomized study of 80 implant acceptors, selected serum lipids were tested before implant insertion and at 6, 12 and 24 months after implant insertion. The lipid parameters evaluated were: total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), triglycerides, apolipoprotein A-I and apolipoprotein B. During the 2 years, the serum lipid pattern in the Implanon users was not significantly different from that of the Norplant users. There was slight decrease in HDL-C levels in both the groups but there were no significant changes in the HDL/TC ratio and the HDL/LDL ratio. Although it was statistically significant, the magnitude of decrease in HDL-C from preinsertion levels in Implanon users was only 5.8% at the end of 2 years. PMID- 14561540 TI - Pharmacokinetic differences between Caucasian and Japanese subjects after single and multiple doses of a potential combined oral contraceptive (Org 30659 and EE). AB - OBJECTIVES: To compare the pharmacokinetic parameters and safety of the progestagen, Org 30659, (17alpha)-17-hydroxy-11-methylene-19-norpregna-4,15-dien 20-yn-3-one), and ethinyl estradiol (EE) in Caucasian and Japanese women after single and multiple doses. METHODS: This was an open-label parallel design of a single dose followed by a multiple dose period in healthy young Japanese and Caucasian subjects. RESULTS: The area under the curve (AUC) of Org 30659 after single dosing was increased by a factor of 1.75 [90% confidence interval (CI): 1.48-2.08] in Japanese women compared to Caucasian women. At steady state, this difference increased to a factor of 1.90 (90% CI: 1.60-2.25). The AUC of EE after single dosing was similar in Caucasian and Japanese women, but at steady state it was increased by a factor 1.38 (90% CI: 1.15-1.64) in the Japanese group. Weight normalization reduced, but did not remove, all the observed differences. Sex hormone binding globulin played no significant role in the differences between Caucasian and Japanese subjects. Both the single- and multiple-dose treatments with Org 30659/EE were generally well tolerated by all subjects. The Japanese population reported more and different treatment-related adverse events than the Caucasian population. CONCLUSIONS: The peak concentration and extent of exposure of Org 30659, and to a lesser extent of EE, in Japanese women are higher than in Caucasian women. Furthermore, the peak concentration and extent of exposure at steady state of Org 30659, and to a lesser extent of EE, are higher than would be predicted assuming linear pharmacokinetics over time. No major safety issues were observed. PMID- 14561541 TI - A single dose of mifepristone (200 mg) in the immediate preovulatory phase offers contraceptive potential without cycle disruption. AB - A single dose of mifepristone is an effective emergency contraceptive and has potential as a regular "once-a-month" pill. If given in the early luteal phase, the formation of a secretory endometrium is inhibited or delayed and implantation of the embryo prevented. We have explored the effect of giving the mifepristone just prior to ovulation on the ovarian and endometrial cycle. Seven women with regular menstrual cycles were studied during a control cycle and then in a second cycle when 200 mg mifepristone was given within 24 h of ovulation, i.e., when luteinizing hormone (LH) in serum was >15 IU/L and the dominant follicle was >18 mm. Ovulation was confirmed within 48 h by ultrasound in five of the seven women. The remaining two women had luteinized unruptured follicle. Following mifepristone, menses occurred after a normal luteal phase compared to control cycle (13.7 +/- 0.7 vs. 13.7 +/- 0.9 days). In all subjects the endometrium on LH + 6 in the treatment cycle showed no, or very little, secretory changes, suggesting it was unlikely that pregnancy would have occurred. We conclude that mifepristone could be given as a "once-a-month" contraceptive pill without causing significant disruption in the menstrual cycle in the majority of women for a 4-day period from just prior to ovulation until LH + 3. PMID- 14561543 TI - Curcumin: a potential vaginal contraceptive. AB - The purpose of this investigation was to evaluate the sperm-immobilizing effects of curcumin, a plant-derived diferuloylmethane compound. Washed human healthy sperm were suspended in Ham's F10 and exposed to varying concentrations of curcumin. Sperm motility was evaluated and changes in sperm mitochondrial transmembrane potential (MTP) was quantified by flow cytometry. Incubation of normal human sperm with curcumin resulted in a dose- and time-dependent loss of sperm motility. At lower concentrations (30 g/mL), curcumin produced a significant (20%) decrease in sperm motility within 30 min without significant effects on sperm viability. An instantaneous (>50%) loss of sperm motility was observed with higher concentrations (300 g/mL) of curcumin and a total loss of sperm motility was achieved within 60 min. A significant reduction in sperm MTP was found with all doses of curcumin tested. Our results indicate that curcumin has a selective sperm-immobilizing effect, in addition to a previously studied anti-HIV property. This compound may have potential clinical applications as a novel intravaginal spermicidal agent for contraception and HIV prevention. PMID- 14561542 TI - A randomized comparison of propofol and methohexital as general anesthetics for vacuum abortion. AB - The objective of this study was to determine whether propofol and methohexital differ in their efficacy, acceptability, cost and side effects when used as the single anesthetic agent for inducing general anesthesia in first-trimester vacuum abortion. We randomized 400 unpremedicated American Society of Anesthesiologists grade I-II women to receive 1% propofol or 1% methohexital by intravenous bolus infusion during abortion. No ancillary anesthetic or adjunctive drugs were used. Visual analogue scale scores were used to grade preoperative patient anxiety and the quality of the anesthetic experience. All personnel outside the operating suites, except the data analyst, were blinded. Chi2, median, analysis of variance and Student's t-tests were used, as appropriate. There were no statistically significant differences in age, gravidity, parity, spontaneous and induced abortion, body mass index or gestational age. Mean total operating time and total drug dose were similar. Propofol use resulted in a significant reduction in the incidence of postoperative nausea (11.5% vs. 17.5%) or vomiting (3.5% vs. 7.0%) with a combined p (mean difference) = 0.002 (95% confidence interval of the mean difference 1.8%, 17.2%). Both the nursing and patient appraisals of the quality of the anesthetic experience favored propofol, but only the nursing scores reached statistical significance (p < 0.001). The cost differential was more than twofold greater for propofol patients, 9.51 dollars vs. 4.42 dollars. In our population of midwestern patients in a major urban area, propofol use had modest advantages over methohexital when used as single agents as judged by first recovery charge nurses, but patients found them equally acceptable. PMID- 14561544 TI - Spermicidal activity of Azadirachta indica (neem) leaf extract. AB - The present study was carried out to evaluate the effective concentration of aqueous extract of old and tender Azadirachta indica (neem) leaves to immobilize and kill 100% human spermatozoa within 20 s. Sander-Cramer test was used to study the spermicidal activity of neem leaf extract. Under the test conditions, minimum effective spermicidal concentrations for tender and old leaf extracts were 2.91 +/- 0.669 mg/million sperm and 2.75 +/- 0.754 mg/million sperm, respectively. The effect of extracts on morphology and viability of sperm was also studied and no change was observed in morphology of head, mid-piece and tail and no viable sperm seen. The leaf extracts were found to be water soluble and carbohydrate in nature. The effect of different concentrations of extracts (old and tender) on percentage motility of the sperm was also studied. With an increase in concentration, there is a linear decrease in percentage motility, becoming zero at a 3-mg dose within 20 s. PMID- 14561546 TI - A generic framework for the parcellation of the cortical surface into gyri using geodesic Voronoi diagrams. AB - In this paper we propose a generic automatic approach for the parcellation of the cortical surface into labeled gyri. These gyri are defined from a set of pairs of sulci selected by the user. The selected sulci are first automatically identified in the data, then projected onto the cortical surface. The parcellation stems from two nested Voronoi diagrams computed geodesically to the cortical surface. The first diagram provides the zones of influence of the sulci. The boundary between the two zones of influence of each selected pair of sulci stands for a gyrus seed. A second diagram yields the gyrus parcellation. The distance underlying the Voronoi diagram allows the method to interpolate the gyrus boundaries where the limiting sulci are interrupted. The method is illustrated with 12 different hemispheres. PMID- 14561547 TI - Statistical shape analysis using non-Euclidean metrics. AB - The contribution of this paper is the adaptation of data driven methods for non Euclidean metric decomposition of tangent space shape coordinates. The basic idea is to extend principal component analysis (PCA) to take into account the noise variance at different landmarks and at different shapes. We show examples where these non-Euclidean metric methods allow for easier interpretation by decomposition into meaningful modes of variation. The extensions to PCA are based on adaptation of maximum autocorrelation factors and the minimum noise fraction transform to shape decomposition. A common basis of the methods applied is the assessment of the annotation noise variance at individual landmarks. These assessments are based on local models or repeated annotations by independent operators. We show that the Molgedey-Schuster independent component analysis is equivalent to the maximum autocorrelation factors. Finally, the different subspace methods are compared using a probabilistic formulation based on their ability to represent the data. PMID- 14561548 TI - Growth modeling of human mandibles using non-Euclidean metrics. AB - From a set of 31 three-dimensional computed tomography (CT) scans we model the temporal shape and size of the human mandible for analysis, simulation, and prediction purposes. Each anatomical structure is represented using 14851 semi landmarks, and mapped into Procrustes tangent space. Exploratory subspace analyses are performed leading to linear models of mandible shape evolution in Procrustes space. The traditional variance analysis results in a one-dimensional growth model. However, working in a non-Euclidean metric results in a multimodal model with uncorrelated modes of biological variation related to independent component analysis. The applied non-Euclidean metric is governed by the correlation structure of the estimated noise in the data. The generative models are compared, and evaluated on the basis of a cross validation study. The new non Euclidean analysis is completely data driven. It not only gives comparable results w.r.t. previous studies of the mean modeling error, but seems to better correlate to growth, and in addition provides the data analyst with alternative hypothesis of plausible shape evolution; hence aiding in the understanding of cranio-facial growth. PMID- 14561549 TI - In vivo strain and stress estimation of the heart left and right ventricles from MRI images. AB - Mechanical properties of the myocardium have been investigated intensively in the last four decades. Many complex strain energy functions have been used to estimate the stress-strain relationship of myocardium because the heart muscle is an inhomogeneous, anisotropic, and nearly incompressible material, which undergoes large deformations. These functions can be effective for fitting in vitro experimental data from myocardial stretch testing. However, it is difficult to model in vivo myocardium using these strain energy functions. Moreover, such estimates have so far been carried out almost exclusively on the left ventricle, because of the relative thinness and complex geometry of the right ventricle. Previous work from our research group has successful estimated the motion and deformation of both the left and the right ventricles, using data from noninvasive tagged magnetic resonance imaging. In this paper, we present a novel statistical model to estimate the in vivo material properties and strain and stress distribution in both ventricles, using such data. Two normal hearts and two hearts with right-ventricular hypertrophy (RVH) were studied and noticeable differences were found between the strain and stress distributions for normal volunteers and RVH patients. Compared to the strain energy function approach, our model is more intuitively understandable. PMID- 14561550 TI - Stochastic finite element framework for simultaneous estimation of cardiac kinematic functions and material parameters. AB - A stochastic finite element framework is presented for the simultaneous estimation of the cardiac kinematic functions and material model parameters from periodic medical image sequences. While existing biomechanics studies of the myocardial material constitutive laws have assumed known tissue kinematic measurements, and image analysis efforts on cardiac kinematic functions have relied on fixed constraining models of mathematical or mechanical nature, we illustrate through synthetic data that a probabilistic joint estimation strategy is needed to achieve more robust and accurate analysis of the kinematic functions and material parameters at the same time. For a particular a priori constraining material model with uncertain subject-dependent parameters and a posteriori noisy imaging based observations, our strategy combines the stochastic differential equations of the myocardial dynamics with the finite element method, and the material parameters and the imaging data are treated as random variables with known prior statistics. After the conversion to state space representation, the extended Kalman filtering procedures are adopted to linearize the equations and to provide the joint estimates in an approximate optimal sense. The estimation bias and convergence issues are addressed, and we conclude experimentally that it is possible to adopt this biomechanical model based multiframe estimation approach to achieve converged estimates because of the periodic nature of the cardiac dynamics. The effort is validated using synthetic data sequence with known kinematics and material parameters. Further, under linear elastic material model, estimation results using canine magnetic resonance phase contrast image sequences are presented, which are in very good agreement with histological tissue staining results, the current gold standards. PMID- 14561551 TI - Spatio-temporal directional filtering for improved inversion of MR elastography images. AB - Dynamic magnetic resonance elastography can visualize and measure propagating shear waves in tissue-like materials subjected to harmonic mechanical excitation. This allows the calculation of local values of material parameters such as shear modulus and attenuation. Various inversion algorithms to perform such calculations have been proposed, but they are sensitive to areas of low displacement amplitude (and hence low SNR) that result from interference patterns due to reflection and refraction. A spatio-temporal directional filter applied as a pre-processing step can separate interfering waves so they can be processed separately. Weighted combinations of inversions from such directionally separated data sets can significantly improve reconstructions of shear modulus and attenuation. PMID- 14561552 TI - Deformable biomechanical models: application to 4D cardiac image analysis. AB - This article describes a methodology for creating a generic volumetric biomechanical model from different image modalities and segmenting time series of medical images using this model. The construction of such a generic model consists of three stages: geometric meshing, non-rigid deformation of the mesh in images of various modalities, and image-to-mesh information mapping through rasterization. The non-rigid deformation stage, which relies on a combination of global and local deformations, can then be used to segment time series of images, e.g. cine MRI or gated SPECT cardiac images. We believe that this type of deformable biomechanical model can play an important role in the extraction of useful quantitative local parameters of cardiac function. The biomechanical model of the heart will be coupled with an electrical model of another collaborative project in order to simulate and analyze a larger class of pathologies. PMID- 14561553 TI - From colour to tissue histology: Physics-based interpretation of images of pigmented skin lesions. AB - Through an understanding of the image formation process, diagnostically important facts about the internal structure and composition of pigmented skin lesions can be derived from their colour images. A physics-based model of tissue colouration provides a cross-reference between image colours and the underlying histological parameters. It is constructed by computing the spectral composition of light remitted from the skin given parameters specifying its structure and optical properties. The model is representative of all the normal human skin colours, irrespective of racial origin, age or gender. Abnormal skin colours do not conform to this model and thus can be detected. Once the model is constructed, for each pixel in a colour image its histological parameters are computed from the model. Represented as images, these 'parametric maps' show the concentration of dermal and epidermal melanin, blood and collagen thickness across the imaged skin as well as locations where abnormal colouration exists. In a clinical study the parametric maps were used by a clinician to detect the presence of malignant melanoma in a set of 348 pigmented lesions imaged using a commercial device, the SIAscope. Logistic regression identified the presence of melanin in the dermis, the abnormal distribution of blood within the lesion and the lesion size as the most diagnostically informative features. Classification based on these features showed 80.1% sensitivity and 82.7% specificity in melanoma detection. PMID- 14561554 TI - Detecting glaucomatous wedge shaped defects in polarimetric images. AB - Wedge shaped defects of the retinal nerve fiber layer (RNFL) may occur in glaucoma. Currently, automatic detection of wedge shaped defects in Scanning Laser Polarimetry (SLP) images of the RNFL is not available. An automatic classification is currently based only on global parameters, thereby ignoring important local information. Our method works by a modified dynamic programming technique that searches for locally strong edges with a preference for straight edges. These edges are initially classified based on their strength and next combined into wedge shaped defects. Our method yields a sensitivity of 73% and a specificity of 90% on a limited set of 65 images. PMID- 14561555 TI - A fully automatic and robust brain MRI tissue classification method. AB - A novel, fully automatic, adaptive, robust procedure for brain tissue classification from 3D magnetic resonance head images (MRI) is described in this paper. The procedure is adaptive in that it customizes a training set, by using a 'pruning' strategy, such that the classification is robust against anatomical variability and pathology. Starting from a set of samples generated from prior tissue probability maps (a 'model') in a standard, brain-based coordinate system ('stereotaxic space'), the method first reduces the fraction of incorrectly labeled samples in this set by using a minimum spanning tree graph-theoretic approach. Then, the corrected set of samples is used by a supervised kNN classifier for classifying the entire 3D image. The classification procedure is robust against variability in the image quality through a non-parametric implementation: no assumptions are made about the tissue intensity distributions. The performance of this brain tissue classification procedure is demonstrated through quantitative and qualitative validation experiments on both simulated MRI data (10 subjects) and real MRI data (43 subjects). A significant improvement in output quality was observed on subjects who exhibit morphological deviations from the model due to aging and pathology. PMID- 14561556 TI - Combinative multi-scale level set framework for echocardiographic image segmentation. AB - In the automatic segmentation of echocardiographic images, a priori shape knowledge has been used to compensate for poor features in ultrasound images. This shape knowledge is often learned via an off-line training process, which requires tedious human effort and is highly expertise-dependent. More importantly, a learned shape template can only be used to segment a specific class of images with similar boundary shape. In this paper, we present a multi scale level set framework for segmentation of endocardial boundaries at each frame in a multiframe echocardiographic image sequence. We point out that the intensity distribution of an ultrasound image at a very coarse scale can be approximately modeled by Gaussian. Then we combine region homogeneity and edge features in a level set approach to extract boundaries automatically at this coarse scale. At finer scale levels, these coarse boundaries are used to both initialize boundary detection and serve as an external constraint to guide contour evolution. This constraint functions similar to a traditional shape prior. Experimental results validate this combinative framework. PMID- 14561557 TI - MAP MRF joint segmentation and registration of medical images. AB - The problems of segmentation and registration are traditionally approached individually, yet the accuracy of one is of great importance in influencing the success of the other. In this paper, we aim to show that more accurate and robust results may be obtained through seeking a joint solution to these linked processes. The outlined approach applies Markov random fields in the solution of a maximum a posteriori model of segmentation and registration. The approach is applied to synthetic and real MRI data. PMID- 14561558 TI - A similarity measure for nonrigid volume registration using known joint distribution of targeted tissue: application to dynamic CT data of the liver. AB - A similarity measure for nonrigid volume registration with known joint distribution of a targeted tissue is developed to process tissue slide at the boundaries between the targeted and non-targeted tissues. Pre-segmentation of the targeted tissue is unnecessary. This measure is applied to registering volumes acquired at different time-phases in dynamic CT scans of the liver using contrast materials and can be derived for the case where only the joint distribution of the targeted tissue is known. The similarity measure is formulated as a likelihood by introducing a concept termed 'exclusivity condition' and embedded into a cost function for nonrigid registration to be combined with the smoothness term. In addition, a practical method for estimating the joint distribution of the liver from unregistered clinical CT data is described. We demonstrate experimentally that tissue slide is effectively processed by this proposed measure using simulated dynamic CT data generated from a software phantom and clinical CT data of eight patients. PMID- 14561559 TI - A viscous fluid model for multimodal non-rigid image registration using mutual information. AB - We propose a multimodal free-form registration algorithm based on maximization of mutual information. The warped image is modeled as a viscous fluid that deforms under the influence of forces derived from the gradient of the mutual information registration criterion. Parzen windowing is used to estimate the joint intensity probability of the images to be matched. The method is evaluated for non-rigid inter-subject registration of MR brain images. The accuracy of the method is verified using simulated multi-modal MR images with known ground truth deformation. The results show that the root mean square difference between the recovered and the ground truth deformation is smaller than 1 voxel. We illustrate the application of the method for atlas-based brain tissue segmentation in MR images in case of gross morphological differences between atlas and patient images. PMID- 14561561 TI - Influence of environment on speech-sound discrimination: findings from a longitudinal study. AB - Event-related potentials (ERPs) from 134 children were obtained at 3 and 8 years of age and recorded to a series of consonant-vowel speech syllables and their nonspeech analogues. The HOME inventory was administered to these same children at 3 and 8 years of age and the sample was divided into 2 groups (low vs. high) based on their HOME scores. Discriminant functions analyses using ERP responses to speech and non-speech analogues successfully classified HOME scores obtained at 3 and 8 years of age and discriminated between children who received low vs. high levels of stimulation for language and reading. PMID- 14561562 TI - The importance of parenting during early childhood for school-age development. AB - This study reports on the relation of 4 patterns of responsive parenting across early childhood (birth-4 1.2 years) with children's rate of cognitive and social development through 8 years of age for a cohort of children born preterm and of very low birthweight and term. Consistent responsiveness across early childhood predicted faster rates of cognitive and social growth than did inconsistent or minimal responsiveness, particularly for preterm children. As parenting at 6 and 8 years did not alter this relation, results suggest that parenting across early childhood plays a unique role in children's development. We also examined whether mothers could be facilitated to use a set of responsive behaviors across infancy that increased children's learning. Compared to control mothers, mothers randomly assigned to receive the intervention displayed greater changes in behaviors that established a social learning context supporting infants' object exploration and vocalization of interests. This, in turn, resulted in greater changes in infants' cognitive and social skills. As in the 1st study, preterm infants appeared to benefit to a greater extent from this more responsive social learning context. PMID- 14561563 TI - Brain mechanisms for reading in children with and without dyslexia: a review of studies of normal development and plasticity. AB - In this article we review our experience with the application of magnetic source imaging (MSI), the newest of the functional imaging methods, to the study of brain mechanisms for reading among children who read normally and among those with dyslexia. After giving a general description of MSI, we present evidence for reliable and valid maps of the brain mechanism for aural language comprehension as well as for reading. Next, we present data from 39 normal readers, 40 children with dyslexia, and 30 younger children at risk for developing a reading disability. These data show different brain activation maps for individual children with dyslexia and children at risk for dyslexia than for those of normal readers. Such differences most likely reflect aberrant brain organization underlying phonological decoding, rather than variables such as degree of effort. Finally, we present preliminary data demonstrating that the aberrant activation profiles of children with dyslexia may return to normative patterns as a result of a successful reading intervention that enables children to improve phonological decoding skills. PMID- 14561564 TI - Interventions aimed at improving reading success: an evidence-based approach. AB - There is a consensus among researchers about the critical elements for effective reading instruction. These elements are the integration of explicit instruction in the alphabetic principle, reading for meaning, and opportunity to learn. These critical elements are present in classroom instruction that prevents reading difficulties as well as effective small-group and one-on-one interventions. Research on effective classroom instruction and reading interventions is described, and the case is argued that the most effective intervention is provided early--in kindergarten through 2nd grade--rather than after 3rd grade, and allows for sufficient intensity, duration, and supportiveness that no child is left behind. Policy implications for changes in (a) the way learning disabilities are identified and (b) the content of professional development of teachers are discussed. PMID- 14561565 TI - Exploring developmental change in the neural bases of higher cognitive functions: the promise of functional magnetic resonance imaging. AB - The dramatic changes in cognitive ability observed throughout childhood mirror comparably significant changes in the developing brain. Studies of animals provide important data on associations between the development of behavior and the neural substrate. However, understanding the development of brain-behavior relations for higher cognitive functions in humans requires direct, concurrent measurement of behavior and brain functions in the children themselves. To date, such data have been very limited. Recent developments in functional magnetic resonance imaging (fMRI) provide the opportunity to systematically explore the development of brain-behavior relations in children. In this article we consider the potential of fMRI to contribute to researchers' understanding of the development of brain-behavior relations. We begin with an overview of the basic imaging method. We then review work from our own laboratory that demonstrates systematic patterns of association between performance on visuospatial tasks and patterns of brain activation, and we compare our findings with those from other laboratories focused on other cognitive domains. Finally, we discuss the potential impact of functional imaging on researchers' understanding of core issues in cognitive and brain development. PMID- 14561566 TI - Early brain injury in children: development and reorganization of cognitive function. AB - Children who sustain congenital or acquired brain injury typically experience a diffuse insult that impacts many areas of the brain. Yet research has only recently begun to examine the development of these children, who often provide excellent examples of the presence or absence of neural plasticity. Development and recovery after such injuries reflects both restoration and reorganization of cognitive functions. To understand these processes, research should focus on questions and assessment paradigms oriented toward the acquisition (rather than the recovery) of cognitive functions. Outcomes may appear similar across types of insults, even when the sources of difficulties and their neural correlates are different. Comparisons of outcomes involving intellectual functions, memory and learning, reading, and language/discourse in children who sustain congenital injury (spina bifida meningomyelocele) and acquired injury (traumatic brain injury) illustrate these principles and the value of research on diffuse brain injury in children. PMID- 14561567 TI - "Rules" of evidence in assessing the efficacy and effectiveness of treatments. AB - A brief review of the "rules" of evidence in assessing the efficacy and effectiveness of treatments leads to a discussion of what the future might hold in the development of the rules of randomized clinical trials (RCTs) in order to enhance the impact of such studies. The primary requisite for enhancing the impact of studies appears to be discussions and interactions among researchers, clinicians, policy makers, and patient advocates, on the one hand, and among the basic scientists and clinical researchers, on the other. PMID- 14561568 TI - Health and human development: understandings from life-course research. AB - It is now well understood that life-course factors affect a diverse range of outcomes, from general well-being to physical functioning and chronic diseases. Exposure to both beneficial and adverse circumstances over the life course will vary for each individual and will constitute a unique life exposure trajectory, which will manifest as different expressions of health and well-being. Here, we present a 3-fold model of life-course influences on health: latency, cumulative, and pathway. By latency we mean relationships between an exposure at one point in the life course and the probability of health expressions years or decades later, irrespective of intervening experience. Cumulative refers to multiple exposures over the life course whose effects on health combine. Finally, pathways represent dependent sequences of exposures in which exposure at 1 stage of the life course influences the probability of other exposures later in the life course, as well as associated expressions. Evidence demonstrating these relationships suggests that, without a consideration of early life as well as adult life experience, policies designed to improve health status tend to overlook root causes. PMID- 14561569 TI - Effects of differential experience on the brain and behavior. AB - This review of plasticity of brain and behavior ranges from examples of plasticity restricted to an early critical period to other examples that show lifelong plasticity, but these two polar opposites are far from covering the field. Rather, it appears that there are a number of patterns by which different measures of plasticity vary as a function of age. Even in cases that indicate the existence of a critical period for development of neural circuits and of behavior, it is not yet possible to define the period exactly, because different methods yield different results, and much parametric work remains to be done. Nevertheless, the variety of findings of plasticity are encouraging for education and for recovery of function. PMID- 14561570 TI - [18F]fluorodeoxyglucose triple-head coincidence imaging as an adjunct to 131I scanning for follow-up of papillary thyroid carcinoma. AB - OBJECTIVE: To evaluate the feasibility of using [(18)F]fluorodeoxyglucose ((18)FDG) triple-head coincidence imaging as a potential cost-effective alternative to positron emission tomography in the setting of suspected recurrence of papillary thyroid carcinoma. METHODS: We retrospectively studied 10 patients with suspected recurrence of papillary carcinoma of the thyroid, who underwent (18)FDG coincidence imaging,(131)I scanning, and a reference anatomic scan (computed tomography, magnetic resonance imaging, or both) within 1 year in most cases. RESULTS: The (131)I scan detected the recurrence in five patients (62.5%) and failed to reveal recurrent cancer in three patients (37.5%); in contrast,(18)FDG imaging detected the recurrence in eight patients (100%) and was true negative in two patients in whom the scans were performed more than 1 year after effective therapy for the recurrence. The sensitivity of detection was unrelated to lesion size. The (18)FDG imaging results led to additional radiotherapy in all (131)I-negative patients, two of whom had high thyroglobulin levels and one of whom had a low thyroglobulin concentration but the presence of antithy-roglobulin antibodies. CONCLUSION: We conclude that (18)FDG triple-head coincidence imaging is useful for routine management of patients with thyroid cancer who have no abnormalities detected on (131)I scans but have high serum thyroglobulin levels. This technique, however, may not be as sensitive as a dedicated positron emission tomographic device, particularly for the assessment of small tumors. PMID- 14561571 TI - Symptomatic and scintigraphic improvement after intravenous pamidronate treatment of Paget's disease of bone in patients with normal serum alkaline phosphatase levels. AB - OBJECTIVE: To describe three patients with symptomatic Paget's disease of bone who presented with normal levels of serum alkaline phosphatase. METHODS: We present three cases of Paget's disease of bone and chronicle the laboratory, scintigraphic, and clinical findings relative to treatment with intravenously administered pamidronate. RESULTS: Although measurement of serum total alkaline phosphatase usually provides a general indication of bone turnover in Paget's disease, about 15% of patients present with normal serum alkaline phosphatase levels. Nonetheless, these patients may have active Paget's disease when assessed with bone scintigraphy or urinary markers of bone resorption. All three study patients had xray findings characteristic of Paget's disease of bone, increased uptake of radiotracer material on bone scans, and elevated levels of urinary markers of bone resorption but normal alkaline phosphatase levels. They were treated with intravenously administered pamidronate, 60 mg once weekly for 2 to 3 consecutive weeks. After treatment, the serum alkaline phosphatase level decreased by 19 to 36%, markers of bone resorption normalized, bone scans showed improvement, and bone pain resolved. CONCLUSION: Pagetic activity in bone and related clinical manifestations may be present in the setting of a normal serum alkaline phosphatase level. Appropriate therapy should not be withheld because of the normal alkaline phosphatase. PMID- 14561572 TI - Potential absence of prognostic implications of severe preoperative hypercalcitoninemia in medullary thyroid carcinoma. AB - OBJECTIVE: To evaluate preoperative hypercalcitonine-mia further as a marker of prognosis in patients with medullary thyroid carcinoma (MTC). METHODS: We reviewed the clinical and laboratory data in six patients (four men and two women, 39 to 76 years old)--three with sporadic MTC, one with familial MTC, and two with multiple endocrine neoplasia type 2A--who had preoperative basal serum calcitonin levels of 400 to 16,000 pg/mL (normal, 0 to 19). Pentagastrin stimulation was performed in patients who had preoperative basal calcitonin levels less than 1,000 pg/mL, and responses ranged from 2,600 to 8,500 pg/mL. Thyroidectomy revealed intrathyroidal MTC in four patients; MTC and nodal metastatic lesions were present in two. The tumor cells were immunoreactive with anti-calcitonin immunoperoxidase staining. RESULTS: Serum calcitonin and carcinoembryonic antigen levels were normal postoperatively. In serial postoperative evaluation during a follow-up period of 2 to 9 years, stimulated peak plasma calcitonin levels after pentagastrin or calcium infusion were normal (in five patients) or near normal (in one patient), without clinical evidence of recurrent disease. The two patients with nodal metastatic disease have had normal calcitonin levels during a mean duration of follow-up of approximately 3 years. CONCLUSION: Pronounced preoperative hypercalci-toninemia does not necessarily preclude a favorable short-term outcome in patients with MTC. PMID- 14561573 TI - Potential association of thyrotoxicosis with vitamin B and folate deficiencies, resulting in risk for hyperhomocysteinemia and subsequent thromboembolic events. AB - OBJECTIVE: To describe a patient with severe thyrotoxicosis attributable to Graves' disease who had a thrombotic cerebrovascular accident and hyperhomocysteinuria, which resolved on correction of the thyrotoxicosis, and to present findings in a pilot study undertaken to investigate the relationship among thyrotoxicosis, homocysteine, folate, and vitamin B(12). METHODS: We present a case report of the index case, with clinical and laboratory details. For the investigative analysis, 21 patients who were 18 to 50 years old and had newly diagnosed, untreated Graves' disease and 10 age-and sex-matched euthyroid control subjects were studied. Of the patients with Graves' disease, 11 underwent studies both at diagnosis and after treatment. Fasting blood tests were performed for thyrotropin, free thyroxine, homocys-teine, vitamin B(12), folate, and methylmalonic acid, a marker of vitamin B(12) deficiency. RESULTS: Vitamin B(12), folate, homocysteine, and methylmalonic acid levels were not significantly different between the thyrotoxic and control or posttreatment groups. In patients with thyrotoxicosis, however, free thyroxine was positively correlated with both homocysteine (r = 0.67; P = 0.03) and methylmalonic acid (r = 0.89; P = 0.003). CONCLUSION: The positive correlation between free thyroxine levels and both homocysteine and methylmalonic acid suggests that thyrotoxicosis may be associated with functional vitamin B(12) deficiency. Such a deficiency may result in clinically important hyperhomocysteine-mia. PMID- 14561574 TI - Panhypopituitarism as an initial manifestation of primary central nervous system non-Hodgkin's lymphoma. AB - OBJECTIVE: To report an unusual case of primary central nervous system non Hodgkin's lymphoma in which the initial manifestation was panhypopituitarism. METHODS: We present a retrospective case review and discuss similar cases from the literature. RESULTS: A 64-year-old woman with nausea, vomiting, diarrhea, and peripheral and periorbital edema was found to have panhypopituitarism. Magnetic resonance imaging showed minimal enlargement of the pituitary, and a transsphenoidal biopsy of the pituitary was nondiagnostic. Months later, abnormalities of extraocular movements developed. Repeated imaging and a second transsphe-noidal biopsy did not reveal the ultimate diagnosis. When further neurologic signs and symptoms subsequently developed, a right temporal open craniotomy was performed. It was not until this procedure, the patient's third biopsy, that the cause of her illness was discovered to be diffuse large cell lymphoma. CONCLUSION: Although idiopathic panhypopituitarism is a relatively common clinical entity, it remains a diagnosis of exclusion. The development of associated neurologic signs should prompt the clinician to initiate a new search for an underlying cause. This case underscores the protean manifestations of central nervous system lymphoma, both endocrine and neurologic, and the difficulties that may be encountered in attempts to establish a diagnosis. PMID- 14561575 TI - Development of diabetes mellitus several years after manifestation of diabetic nephropathy: case report and review of literature. AB - OBJECTIVE: To describe an unusual case of development of diabetes mellitus (DM) several years after manifestation of diabetic nephropathy and to review the related literature. METHODS: We present a case report, including detailed laboratory and pathologic findings in a 51-year-old man who was diagnosed as having DM several years after presenting with diabetic nephropathy. The pertinent literature is also reviewed. RESULTS: A 51-year-old African American man presented with proteinuria of 4 g/24 h. Past medical history was significant for impaired glucose tolerance diagnosed 2 years previously. Subsequent follow-up demonstrated fasting blood glucose levels ranging from 108 to 123 mg/dL and glycated hemoglobin levels ranging from 5.3 to 5.8%. The patient also had chronic hepatitis C, hypertension, a history of intravenous drug abuse, and a family history of DM and hypertension. On examination of the patient, his blood pressure was 180/90 mm Hg. Funduscopy revealed mild diabetic retinopathy. Work-up was negative for glomerulonephritis, connective tissue disease, vasculitis, or multiple myeloma. Kidney biopsy revealed thickened glomerular basement membranes and diffuse glomeru-losclerosis, consistent with diabetic nephropathy. During follow-up, 9 years after presenting with proteinuria and 4 years after diagnosis of biopsy-proven diabetic nephropathy, the patient had a blood glucose level of 890 mg/dL and diabetic ketoacidosis. CONCLUSION: This case provides one explanation for the natural course of patients who present with "diabetic complications" but have no diabetes. Some of those patients may have "prediabetes" and may manifest with DM during follow-up. We also conclude that hyperglycemia is not the only important factor in the pathogenesis of diabetic nephropathy. PMID- 14561576 TI - Metformin-induced cholestatic hepatitis. AB - OBJECTIVE: To report a case of metformin-induced cholestatic hepatitis. METHODS: We present a detailed case report, including laboratory and biopsy findings. In addition, similar cases from the literature are reviewed. RESULTS: In a 68-year old man with newly diagnosed diabetes mellitus, metformin therapy was begun. The dosage initially was 500 mg twice daily and later was increased to 850 mg twice a day. Four weeks after met-formin treatment was initiated, jaundice, pruritus, and liver enzyme abnormalities were noted. The patient underwent an extensive work up, including a hepatitis screen, ultrasonography, magnetic resonance imaging, and endoscopic retrograde cholangiopancreatography, all of which showed normal findings. A liver biopsy revealed severe cholestasis and mild portal inflammation. Treatment with metformin was discontinued, and the liver enzymes normalized except for a persistently increased level of alkaline phosphatase, most likely related to a prolonged cholestatic effect of metformin. CONCLUSION: Although rare, metformin can be responsible for inducing liver damage, and patients and physicians should be aware of this side effect. PMID- 14561578 TI - Skeletal effects of statins. AB - OBJECTIVE: To review the skeletal effects of hydroxy-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors (statins) and evaluate the use and potential benefit of statins in the management of osteoporosis. METHODS: Background information on the topic of the effect of statins on bone and fracture risk is presented, and the pertinent published literature is reviewed. RESULTS: Osteoporosis is the most common bone disease, affecting millions of people worldwide and leading to considerable morbidity, especially when it is not adequately managed. Although statins have primarily been known for their lipid lowering effects, recent data have documented a potential association between statin use and improvement in fracture risk profile. Some statins have been shown to decrease bone-specific alkaline phosphatase or serum osteocalcin concentrations, but other studies have failed to demonstrate the beneficial effects of these agents. Therefore, additional studies should be undertaken to clarify the mechanism of action. CONCLUSION: Observational studies suggest an association between HMG-CoA reductase inhibitors and reduction in fracture risk. Large randomized controlled clinical trials must be performed to confirm this association. PMID- 14561577 TI - Hypotension in a woman with a metastatic dopamine-secreting carotid body tumor. AB - OBJECTIVE: To describe a woman with metastatic carotid body tumor in whom hypotension occurred in the setting of exceedingly high plasma dopamine levels. METHODS: We present a case report and review the literature on the topic of dopamine-secreting paraganglioma or pheochromocytoma. RESULTS: A previously healthy 40-year-old Asian woman noted difficulty with swallowing and hoarseness. No neck mass was visible, and she had no symptoms of catecholamine excess and no family history of endocrine disorders or malignant disease. Indirect laryngoscopy revealed a paralyzed left vocal cord and a nonulcerating mass in the left parapharyngeal space. An initial needle biopsy was interpreted as undifferentiated carcinoma. After a second biopsy, this mass was diagnosed as a neuroendocrine tumor, consistent with paraganglioma. The patient underwent surgical resection and radiation therapy (total dose, 40 Gy), after which she remained asymptomatic for 11 years. Then loss of weight, fatigue, nausea, and hypotensive episodes (blood pressures as low as 70/35 mm Hg) prompted whole-body imaging with bone scans, computed tomography, and magnetic resonance imaging, which disclosed several lesions in the liver, lungs, and spine, suggestive of metastatic disease. The adrenal glands were unremarkable. A metaiodobenzylguanidine scan with use of (131)I was negative. Liver biopsy of a hypodense lesion revealed a neuroendocrine tumor by histologic and immunohistochemical studies. Because of the patient's history, malignant paraganglioma was diagnosed. The tumor secreted predominantly dopamine at extraordinary levels (plasma concentration 27,942 pg/mL; normal, <30). The patient died before further treatment could be initiated. CONCLUSION: Carotid body tumors usually do not secrete catecholamines but frequently metastasize. During progression, these neuroendocrine tumors may become able to produce and secrete selected catecholamines such as dopamine. Dopamine can lower the blood pressure rather than causing hypertension, even though hypertension is one of the main symptoms of a pheochromocytoma. PMID- 14561579 TI - Evidence-based endocrine practice. PMID- 14561580 TI - Changing paradigms in the follow-up of patients with differentiated thyroid cancer: an alternative to [18F]fluorodeoxyglucose positron emission tomographic scanning. PMID- 14561581 TI - Idle thoughts from an addled mind. Whose interest is being served? PMID- 14561582 TI - Complexity of Langerhans' cell histiocytosis. PMID- 14561583 TI - Visual vignette. PMID- 14561585 TI - Regulation of expression of the intestinal oligopeptide transporter (Pept-1) in health and disease. AB - The abundance of the oligopeptide transporter (Pept-1) in the brush-border membrane of the intestinal epithelium is the central mechanism for regulation of transport of products of protein digestion (dipeptides and tripeptides) and peptidomimetic drugs (for example, beta-lactam antibiotics). Within the past few years, there has been substantial progress in identifying the factors controlling this regulation and the mechanisms of their actions. The purpose of this report is to review this progress. The studies of individual substrates and hormones in a human intestinal cell line (Caco-2) have shown that dipeptides, certain amino acids, insulin, and leptin increase and epidermal growth factor and triiodothyronine decrease the membrane population of Pept-1. In the case of dipeptides, epidermal growth factor, and thyroid hormone, there are parallel changes in the gene expression brought about by alteration of transcription and/or stability of Pept-1 mRNA. In contrast, the treatment with insulin and leptin does not induce any alteration in the Pept-1 gene expression, and the mechanism of increased protein expression appears to be increased trafficking from a preformed cytoplasmic pool to the apical membrane. In vivo studies in rats have shown modulation of protein and gene expressions of the intestinal oligopeptide transporter during the day and during development and in nutritional and metabolic alterations, such as high-protein diet, fasting, and diabetes. Patients with intestinal diseases, such as ulcerative colitis, Crohn's disease, and short-bowel syndrome, may have induction of the Pept-1 expression in their colon. Finally, pharmacological studies have shown that the expression of Pept-1 can be upregulated by agents such as 5 fluorouracil and downregulated by agents such as cyclosporine. In conclusion, the above studies have produced a wealth of new information on regulation of a key transporter in the intestine. This information may have useful applications in nutritional and pharmacological treatments, for example, in diabetic patients needing enteral nutrition or in ulcerative colitis patients needing the suppression of the intestinal inflammation. PMID- 14561586 TI - Differential effects of polyunsaturated fatty acids on sterol synthesis rates in adult and fetal tissues of the hamster: consequence of altered sterol balance. AB - Cholesterol is necessary for the proper growth and development of the fetus. Consequently, disruptions in cholesterol biosynthesis lead to abnormal fetal development. It has been shown that in cells exposed to polyunsaturated fatty acids (PUFA), the expressions of genes and activities of enzymes involved in cholesterol synthesis are reduced. Similarly, we found that adult male hamsters fed PUFA-enriched diets had an approximately 60% reduction in in vivo hepatic sterol synthesis rates. If fetal tissues respond to PUFA in the same manner as do adult livers, then maternal dietary PUFA could lead to a reduction in fetal sterol synthesis rates and possibly abnormal development. To investigate the impact of maternal dietary fatty acids on fetal sterol synthesis rates, female hamsters were fed diets enriched in various fatty acids before and throughout gestation. In vivo sterol synthesis rates were measured in fetuses at mid- and late gestation. At both gestational stages, dietary PUFA had no effect on fetal sterol synthesis rates. This lack of effect was not a consequence of a lack of PUFA enrichment in fetal fatty acids or the lack of PUFA receptor expression in the fetus. We hypothesize that the fetus may experience a dysregulation of sterol synthesis as the result of the fetus being in a negative sterol balance; the PUFA induced suppression of sterol synthesis in the adult male hamster liver was ablated by creating a net negative sterol balance across the adult hepatocyte. PMID- 14561588 TI - Flow fields generated by peristaltic reflex in isolated guinea pig ileum: impact of contraction depth and shoulders. AB - The guinea pig ileum responds to distension with characteristic wall movements, luminal pressure gradients, and outflow (the peristaltic reflex). To date, little is known about whether the peristaltic reflex generates flow events other than laminar flow. Here we used a numerical method to solve for the flow generated by moving walls to assess occlusive contractions (case 1), nonocclusive contractions (case 2), and contractions with steep shoulders (case 3) for which visual parameters of wall movements are published. We found that all three contraction cases produced pressure differentials across the coapting segment, downstream and reverse flow, and vortical flow patterns that redistributed particles and mixed liquids. Contractions generated pressures and shear stresses, particularly along the moving section of the wall. The nonocclusive contraction was much less effective than the occlusive contraction with the steep shoulders; the occlusive contraction with flat shoulders had an intermediate effect. Our analysis shows that even peristaltic contractions produce not only laminar flow but also many flow events likely to promote digestion and absorption. The visual patterns of contractions impact the patterns of luminal flow, and precise definition of wall movements is critical to quantify the fluid mechanical consequences of intestinal contractions. PMID- 14561587 TI - Mechanism of active repolarization of inhibitory junction potential in murine colon. AB - Enteric inhibitory responses in gastrointestinal (GI) smooth muscles involve membrane hyperpolarization that transiently reduce the excitability of GI muscles. We examined the possibility that an active repolarization mechanism participates in the restoration of resting membrane potential after fast inhibitory junction potentials (IJPs) in the murine colon. Previously, we showed these cells express a voltage-dependent nonselective cation conductance (NSCC) that might participate in active repolarization of IJPs. Colonic smooth muscle cells were impaled with micro-electrodes and voltage responses to nerve-evoked IJPs, and locally applied ATP were recorded. Ba2+ (500 muM), a blocker of the NSCC, slowed the rate of repolarization of IJPs. We also tested the effects of Ba2+, Ni2+, and mibefradil, all blockers of the NSCC, on responses to locally applied ATP. Spritzes of ATP caused transient hyperpolarization, and the durations of these responses were significantly increased by the blockers of the NSCC. We considered whether NSCC blockers might affect ATP metabolism and found that Ni2+ decreased ATP breakdown in colonic muscles. Mibefradil had no effect on ATP metabolism. Because both Ni2+ and mibefradil had similar effects on prolonging responses to ATP, it appears that restoration of resting membrane potential after ATP spritzes is not primarily due to ATP metabolism. Neurally released enteric inhibitory transmitter and locally applied ATP resulted in transient hyperpolarizations of murine colonic muscles. Recovery of membrane potential after these responses appears to involve an active repolarization mechanism due to activation of the voltage-dependent NSCC expressed by these cells. PMID- 14561589 TI - Hypoxia induces differentiation of pulmonary artery adventitial fibroblasts into myofibroblasts. AB - Activation of the alpha-smooth muscle actin (alpha-SMA) gene during the conversion of fibroblasts into myofibroblasts is an essential feature of various fibrotic conditions. Microvascular compromise and thus local environmental hypoxia are important components of the fibrotic response. The present study was thus undertaken to test the hypothesis that hypoxia can induce transdifferentiation of vascular fibroblasts into myofibroblasts and also to evaluate potential signaling mechanisms governing this process. We found that hypoxia significantly upregulates alpha-SMA protein levels in bovine pulmonary artery adventitial fibroblasts. Increased alpha-SMA expression is controlled at the transcriptional level because the alpha-SMA gene promoter activity, assayed via a luciferase reporter, was markedly increased in transfected fibroblasts exposed to hypoxia. Hypoxic induction of the alpha-SMA gene was mimicked by overexpression of constitutively active Galphai2 (alphai2Q205L) but not Galpha16 (alpha-16Q212L). Blockade of hypoxia-induced alpha-SMA expression with pertussis toxin, a Galphai antagonist, confirmed a role for Galphai in the hypoxia-induced transdifferentiation process. c-Jun NH2-terminal kinase (JNK) inhibitor II and SB202190, but not U0126, also attenuated alpha-SMA expression in hypoxic fibroblasts, suggesting the importance of JNK in the differentiation process. Hypoxia-induced increase in bromodeoxyuridine incorporation, which occurred concomitantly with hypoxia-induced differentiation, was blocked by U0126, suggesting that DNA synthesis and alpha-SMA expression take place through simultaneously activated parallel signaling pathways. Neutralizing antibody against transforming growth factor-beta1 blocked only 30% of the hypoxia-induced alpha-SMA promoter activity. Taken together, our results suggest that hypoxia induces differentiation of vascular fibroblasts into myofibroblasts by upregulating the expression of alpha-SMA, and this increase in alpha-SMA level occurs through Galphai- and JNK-dependent signaling pathways. PMID- 14561590 TI - Rapid muscle-specific gene expression changes after a single bout of eccentric contractions in the mouse. AB - Eccentric contractions (ECs), in which a muscle is forced to lengthen while activated, result in muscle injury and, eventually, muscle strengthening and prevention of further injury. Although the mechanical basis of EC-induced injury has been studied in detail, the biological response of muscle is less well characterized. This study presents the development of a minimally invasive model of EC injury in the mouse, follows the time course of torque recovery after an injurious bout of ECs, and uses Affymetrix microarrays to compare the gene expression profile 48 h after ECs to both isometrically stimulated muscles and contralateral muscles. Torque dropped by approximately 55% immediately after the exercise bout and recovered to initial levels 7 days later. Thirty-six known genes were upregulated after ECs compared with contralateral and isometrically stimulated muscles, including five muscle-specific genes: muscle LIM protein (MLP), muscle ankyrin repeat proteins (MARP1 and -2; also known as cardiac ankyrin repeat protein and Arpp/Ankrd2, respectively), Xin, and myosin binding protein H. The time courses of MLP and MARP expression after the injury bout (determined by quantitative real-time polymerase chain reaction) indicate that these genes are rapidly induced, reaching a peak expression level of 6-11 times contralateral values 12-24 h after the EC bout and returning to baseline within 72 h. Very little gene induction was seen after either isometric activation or passive stretch, indicating that the MLP and MARP genes may play an important and specific role in the biological response of muscle to EC-induced injury. PMID- 14561592 TI - Catheter angiography is still necessary for the measurement of carotid stenosis. PMID- 14561591 TI - Role of cytoplasmic termini in sorting and shuttling of the aquaporin-2 water channel. AB - In mammals, the regulation of water homeostasis is mediated by the aquaporin-1 (AQP1) water channel, which localizes to the basolateral and apical membranes of the early nephron segment, and AQP2, which is translocated from intracellular vesicles to the apical membrane of collecting duct cells after vasopressin stimulation. Because a similar localization and regulation are observed in transfected Madin-Darby Canine Kidney (MDCK) cells, we investigated which segments of AQP2 are important for its routing to forskolin-sensitive vesicles and the apical membrane through analysis of AQP1-AQP2 chimeras. AQP1 with the entire COOH tail of AQP2 was constitutively localized in the apical membrane, whereas chimeras with shorter COOH tail segments of AQP2 were localized in the apical and basolateral membrane. AQP1 with the NH2 tail of AQP2 was constitutively localized in both plasma membranes, whereas AQP1 with the NH2 and COOH tail of AQP2 was sorted to intracellular vesicles and translocated to the apical membrane with forskolin. These data indicate that region N220-S229 is essential for localization of AQP2 in the apical membrane and that the NH2 and COOH tail of AQP2 are essential for trafficking of AQP2 to intracellular vesicles and its shuttling to and from the apical membrane. PMID- 14561593 TI - The promise of high-field-strength MR imaging. PMID- 14561594 TI - Neuroethics in a new era of neuroimaging. PMID- 14561595 TI - The need for a West Nile virus MRI registry. PMID- 14561596 TI - Evaluation of head and neck squamous cell carcinoma after treatment. PMID- 14561597 TI - Accuracy of contrast-enhanced MR angiography in predicting angiographic stenosis of the internal carotid artery: linear regression analysis. AB - BACKGROUND AND PURPOSE: We sought to assess whether contrast-enhanced MR angiography is able to predict the degree of angiographic stenosis of the internal carotid artery within a clinically acceptable margin of error, thereby decreasing the need for angiography. In addition, we sought to assess whether adding ultrasound peak systolic velocity (PSV) as an additional regressor improves the accuracy of prediction. METHODS: A retrospective review of our institution's records for a 4-year period was conducted to identify all patients who had undergone evaluation of their carotid arteries using digital subtraction angiography, contrast-enhanced MR angiography, and ultrasonography. All internal carotid artery stenoses ranging from 10% to 90% at carotid angiography were selected (n = 22). Measurements were then obtained based on the North American Symptomatic Carotid Endarterectomy Trial style by using the digital subtraction angiograms and contrast-enhanced MR angiograms in a blinded fashion. The correlation between digital subtraction angiography data and contrast-enhanced MR angiography data was assessed by conducting linear regression analysis. Multiple regression analysis was then conducted to determine whether the inclusion of ultrasound PSV as an additional regressor increased the accuracy of prediction. RESULTS: The correlation between the degree of stenosis measured by digital subtraction angiography and that measured by contrast-enhanced MR angiography was r = 0.967. The 95% confidence interval for the line of means showed low errors bounds, ranging as low as +/-2.8%. The 95% confidence interval for individual prediction of angiographic stenosis based on a given contrast-enhanced MR angiographic measurement, however, was significantly larger, being no less than +/-13.6%. With the inclusion of PSV, the adjusted correlation was r = 0.965. CONCLUSION: A clear linear relationship exists between digital subtraction angiographic and contrast-enhanced MR angiographic measurements of carotid stenosis. Increasing severity of stenosis as measured by contrast-enhanced MR angiography corresponds to increasing severity at angiography. Although the predictive value of contrast-enhanced MR angiography is excellent in the mean, it is less reliable for predicting the degree of angiographic stenosis in the individual patient, showing rather wide confidence intervals. Furthermore, the inclusion of PSV as an additional regressor does not improve the predictive accuracy beyond that of contrast-enhanced MR angiography alone. PMID- 14561598 TI - Earring lesions of the parotid tail. AB - BACKGROUND AND PURPOSE: Masses in the parotid tail can be a source of consternation to radiologists and clinicians; inaccurate localization may lead to significant iatrogenic complication. We sought to review the pertinent anatomic localizing features of the parotid tail, relevant facial nerve anatomy, and sources of clinical and radiologic confusion. To conclude, we review imaging features that are helpful in generating a diagnosis in this location. METHODS: We retrospectively reviewed the imaging and clinical features of 111 parotid tail masses in 103 patients (56 male, 45 female, two of unknown sex; age range, 5 months-81 years). The following imaging findings were noted: size, enhancement, multiplicity of lesions, attenuation on CT scans, signal intensity on MR images, and appearance of the surrounding parotid gland. Diagnosis was confirmed by either surgical resection or biopsy findings or by specific clinical data or characteristic imaging findings. RESULTS: Seventeen types of parotid tail masses were identified. Benign lesions were: pleomorphic adenoma (n = 15), Warthin tumor (n = 14), infectious process (n = 13), venous malformation (n = 9), and Sjogren disease (n = 9), lymphatic malformations (n = 7), lipoma (n = 6), HIV lymphoepithelial lesion (n = 4), first brachial cleft cyst (n = 3), oncocytoma (n = 2), sarcoid (n = 1), and lymph node (n = 1). Malignant lesions were: Non Hodgkin lymphoma (n = 14), metastatic disease (n = 7), mucoepidermoid carcinoma (n = 4), acinic cell carcinoma (n = 1), and undifferentiated carcinoma (n = 1). Eight patients had two diagnoses. CONCLUSION: Understanding normal parotid tail anatomy is important to radiologists, because accurate localization has implications for appropriate management of masses in this location, potentially reducing the occurrence of marginal mandibular nerve injury. PMID- 14561599 TI - Mesectodermal leiomyosarcoma of the ciliary body. AB - We report a case of low-grade mesectodermal leiomyosarcoma of the ciliary body, an extremely rare tumor of neural crest origin, occurring in a 12-year-old boy. MR imaging showed a well-marginated, ovoid soft tissue mass at the right temporal ciliary body, accompanied by total retinal detachment. The mass was slightly hyperintense relative to contralateral vitreous on T1-weighted images and markedly hypointense on T2-weighted images and enhanced very well. Initial biopsy of the mass suggested a peripheral nerve sheath tumor. The mass grew rapidly after biopsy, and enucleation was performed. The final diagnosis based on histology and immunohistochemistry was a low-grade mesectodermal leiomyosarcoma. To our knowledge, this is the first reported case of a malignant form of mesectodermal leiomyoma of the ciliary body. PMID- 14561600 TI - CT characteristics of intraocular perfluoro-N-octane. AB - Perfluoro-N-octane (PFO) is a heavy liquid that is used as an aid for complicated retinal surgical procedures. Although PFO is usually removed intraoperatively, the radiographic appearance of retained PFO may mimic an intraocular foreign body or vitreous hemorrhage. As the use of PFO in retinal procedures has become more widespread, recognition of its imaging appearance has become important in the differential diagnosis of intraocular foreign body and ocular trauma. PMID- 14561601 TI - Endovascular interventional neuroradiologic procedures: who is performing them, how often, and where? A survey of academic and nonacademic radiology practices. AB - BACKGROUND AND PURPOSE: In this report, the authors assess practice patterns at both academic and nonacademic centers regarding the treatment of aneurysms with Guglielmi detachable coils (GDCs), thrombolysis of the carotid-vertebral arteries, and stent placement with angioplasty of the carotid arteries. METHODS: A neurovascular radiology survey was sent to 102 directors of neuroradiology fellowship programs in the United States and Canada ("academic centers"). The survey was also sent to senior members of the American Society of Neuroradiology (three per state) who were not affiliated with fellowship programs ("nonacademic centers"). RESULTS: Fifty-seven surveys from academic practices and 70 surveys from nonacademic practices were returned. A total of 4361 procedures (2283 GDC; 949 thrombolysis; 1129 stent placement) were performed; 84% were performed at academic centers and 16% at nonacademic centers. Ninety percent of GDC, 71% of thrombolysis, and 82% of stent placement procedures were performed at academic centers. Seven academic and three nonacademic centers performed 48% of all GDC procedures; eight academic and four nonacademic centers performed 45% of all thrombolysis procedures; eight academic centers performed 50% of all stent placement procedures. A total of 544/4361 (12%) procedures were performed by nonradiologists. At academic centers, 14% of procedures were performed by nonradiologists; participation by nonradiologists was greatest for carotid stent placement (24% of procedures). At nonacademic centers, only 5% of procedures were performed by nonradiologists. CONCLUSION: According to this survey, most endovascular interventional neuroradiologic procedures are performed at academic centers; given the survey population, this study likely identifies the lower limit of participation by nonradiologists (12%). Performance of these procedures is concentrated in relatively few centers, and these data raise questions about the overall use of intraarterial thrombolytic therapy for acute infarction. PMID- 14561602 TI - High-concentration ethylene-vinyl alcohol copolymer and endovascular treatment of experimental aneurysms: feasibility of embolization without protection devices at the neck. AB - BACKGROUND AND PURPOSE: Coiling of intracranial aneurysms is both safe and effective but may be followed by recurrences. The purpose of this study was to assess the feasibility of endovascular treatment of aneurysms with high concentration ethylene-vinyl alcohol copolymer (HCEVOH), without the use of protection devices at the neck. METHODS: Wide-necked bifurcation aneurysms with a high propensity for recurrences were constructed in 22 dogs. HCEVOH embolization was performed with a dedicated high-pressure microcatheter in 12 animals. Angiographic results at 3 and 12 weeks and pathologic results at 12 weeks were compared with those of a separate group of 10 animals treated with platinum coils. We used a qualitative scoring system to grade angiographic results, neointima formation, and recanalization at the neck. RESULTS: Intraaneurysmal HCEVOH injections could be performed without carotid emboli and without a protection device in 11 of 12 animals. Fragments detached upon traction of the microcatheters at the end of the procedure on two occasions. Immediate and late angiographic results were not significantly different between the two groups (P =.807), with a tendency for angiographic recurrences at 3 months (angiographic scores were significantly worse in both groups at 12 weeks as compared with T0 [P <.02]). A complete occlusion, including the neck, even at the cost of protrusion of material at the level of the branches, is necessary to decrease risks of recurrences. Neointima formation at the surface of the embolic agent was complete at the neck of aneurysms treated with HCEVOH. The neointimal score was significantly improved with HCEVOH as compared with coil embolization (P =.03). CONCLUSION: HCEVOH embolization of aneurysms without neck protection is feasible. It does not, however, eliminate recurrences in an experimental wide-necked aneurysm model. PMID- 14561603 TI - Endovascular trapping of traumatic carotid-cavernous fistulae. AB - We describe an unconventional endovascular approach to the treatment of traumatic carotid-cavernous fistulas. Four patients with large high-flow shunts have been successfully treated by trapping of the fistula by using a combination of proximal balloon occlusion and distal coil embolization. The latter was achieved following retrograde catheterization of the distal parent vessel via the contralateral carotid or ipsilateral vertebral artery. PMID- 14561604 TI - Endovascular treatment strategy for direct carotid-cavernous fistulas resulting from rupture of intracavernous carotid aneurysms. AB - BACKGROUND AND PURPOSE: Reported treatments and outcomes in aneurysmal carotid cavernous fistulas (CCFs) have been admixed with those of cases considered to be symptomatic of intracavernous aneurysm. However, aneurysmal CCFs have clinical features distinct from those of dural arteriovenous fistulas, and treatment strategies similar to those of traumatic CCF are required. We evaluated our experience in placing detachable balloons in the management of spontaneous CCFs due to rupture of an intracavernous aneurysm. METHODS: Six patients (one man, five women; mean age, 64.7 years) were treated for spontaneous direct CCF at our institution between 1995 and 2001. All patients presented with sudden ocular symptoms including exophthalmos, conjunctival injection, chemosis, and ocular motor palsies. Detachable latex balloons were used as the embolic material in five patients, and in one patient the cavernous sinus was packed transarterially with coils. RESULTS: All six patients were successfully treated by means of transarterial embolization, and symptoms improved within a week. CONCLUSION: Although other techniques using a transvenous approach and/or detachable coils may also be useful, embolization with detachable balloons should be a safe and effective method to immediately occlude the fistula. PMID- 14561605 TI - Intracranial aneurysms treated with Guglielmi detachable coils: is contrast material necessary in the follow-up with 3D time-of-flight MR angiography? AB - BACKGROUND AND PURPOSE: Three-dimensional time-of-flight (TOF) MR angiography has been evaluated in the follow-up of intracranial aneurysms treated with Guglielmi detachable coils (GDCs) with good results. Some of the studies used contrast material in addition to the 3D TOF MR technique and others did not. We assessed the usefulness of contrast material with 3D TOF MR angiography by comparing this sequence before and after contrast material injection. METHODS: Fifty-eight patients harboring a total of 71 cerebral aneurysms previously treated with GDCs were included in the prospective study. MR angiography (at 1.5 T) was performed with a 3D TOF sequence before and after injection of gadolinium-based contrast material. Features evaluated were presence and size of a neck remnant, parent and adjacent vessel patency, and venous overlap. Digital subtraction angiography was the standard of reference. RESULTS: Comparison of the techniques showed a good agreement in the detection of residual flow. Six cases of small residual neck were not detected with either the 3D TOF or the contrast-enhanced 3D TOF sequence. In one case of giant aneurysm, the extent of recanalization was more evident after contrast material administration. The use of contrast material did not help to show the parent and adjacent arteries. Venous overlap on contrast enhanced 3D TOF angiograms did not affect image interpretation. CONCLUSION: In this series, the use of intravenous contrast material did not improve the ability of 3D TOF MR angiography to depict the presence of residual or recurrent aneurysms previously treated with endovascular coiling. In one giant aneurysm, use of intravenous contrast material did result in improved visualization of a residual aneurysm. PMID- 14561606 TI - Computational approach to quantifying hemodynamic forces in giant cerebral aneurysms. AB - BACKGROUND AND PURPOSE: The options for treating giant fusiform basilar aneurysms are limited, and the potential impact of planned interventions is difficult to assess. We developed a computational framework to evaluate the impact that interventions might have on hemodynamic conditions. METHODS: A computational fluid dynamics approach was used to determine the velocity field, wall shear stress, and pressure distribution within a model of a basilar artery before and after a simulated occlusion of one vertebral artery. The vascular geometry in a patient with a giant fusiform basilar artery aneurysm was determined by using contrast-enhanced MR angiography, and the numerical simulation approach was used to calculate the flow fields in the presenting geometry and to predict the flow field that would occur if a vertebral artery were occluded. RESULTS: In the model geometry, computational fluid dynamics indicated that there would be a symmetric flow pattern with a strong central stream and large recirculation zones at the walls. After simulated occlusion of one vertebral artery, the primary stream was diverted to one side, resulting in high pressure and increased wall shear stress. For the patient-specific geometry, flow patterns were shown to depend strongly on how much flow there was in each vertebral artery. CONCLUSION: Contrast-enhanced MR angiography is an effective tool for demonstrating the luminal boundaries of large intracranial aneurysms. Computational fluid dynamics is a powerful tool for determining the prevailing flow conditions in vascular territories and for modeling the possible alterations of the flow field that would result from interventional treatments. PMID- 14561607 TI - De novo aneurysm formation 6 and 22 months after initial presentation in two patients. AB - Two cases of angiographically and surgically documented de novo intracranial aneurysm formation are reported. The first patient, a 32-year-old woman, developed two new aneurysms within a 6-month period, and the second patient, a 27 year-old woman, developed two new aneurysms within a 22-month period. In both patients, the new aneurysms were symptomatic, causing stroke and subarachnoid hemorrhage, respectively. The development of aneurysms de novo over such a short period of time has important implications for periodic imaging in patients considered to be at high risk for intracranial aneurysm formation. PMID- 14561608 TI - Stent-within-a-stent technique for the treatment of dissecting vertebral artery aneurysms. AB - Subarachnoid hemorrhage from dissecting vertebral artery aneurysms is a well known phenomenon. With the advent of navigable intracranial stents, these aneurysms are now amenable to endovascular stent placement. However, immediate aneurysm obliteration is often not accomplished, and current covered stents have poor short-term patency rates. We report a series of three patients with intradural dissecting vertebral artery aneurysms treated with a novel endovascular stent-within-a-stent construct using currently available stent technology. PMID- 14561609 TI - Management of neuroform stent dislodgement and misplacement. AB - A self-expanding stent has recently been introduced for the treatment of wide neck aneurysms. We describe two cases of stent malposition within large aneurysms. In the first case, the stent was dislodged during microcatheterization. This was managed by placement of a second stent through the interstices of the first followed by aneurysm coiling. In the second case, after deployment, the proximal portion of the stent moved into the aneurysm as the exchange guidewire was removed. This patient was treated by vessel sacrifice. PMID- 14561610 TI - Perfusion- and diffusion-weighted MR imaging-guided therapy of vertebral artery dissection: intraarterial thrombolysis through an occipital vertebral anastomosis. AB - Management of arterial dissections can be particularly challenging. We report a case of vertebral artery dissection in which perfusion- and diffusion-weighted MR imaging findings suggested the presence of salvageable tissue, despite that the patient had symptoms for more than 40 hours. Direct access to the distal vascular territory was unattainable, and the presence of collateral circulation through an occipital vertebral anastomosis provided the only pathway to administer intraarterial thrombolysis. This case demonstrates that perfusion- and diffusion weighted MR imaging can be instrumental in the selection of candidates for aggressive stroke therapy. Arterial anastomoses can provide alternate access to ischemic vascular beds and merit careful evaluation during intraarterial thrombolysis. PMID- 14561611 TI - Combined stent placement and thrombolysis in acute vertebrobasilar ischemic stroke. AB - BACKGROUND AND PURPOSE: Acute vertebrobasilar ischemic stroke is often associated with high morbidity and mortality with limited therapeutic options. Endovascular treatment with thrombolysis has offered some hope for affected patients; however, overall outcomes have been less than satisfactory. In this report, we present the results of our approach in six consecutive cases of acute vertebrobasilar ischemic stroke by combined proximal vessel stent placement and thrombolysis. METHODS: Six consecutive cases were retrospectively reviewed for the clinical outcome of patients presenting to our institution with acute posterior circulation stroke who underwent cerebral revascularization including proximal arterial stent placement by using balloon-expandable coronary stents and intraarterial thrombolysis. All of these patients were initially evaluated by stroke team neurologists and imaged with MR, including diffusion-weighted imaging documenting acute posterior circulation stroke. MR angiography of the circle of Willis was also obtained. Short-term follow-up was conducted to assess National Institutes of Health stroke scores (NIHSS) and modified Rankin scores. RESULTS: In these six cases, a combined approach of proximal arterial stent placement (five cases of vertebral artery origin and one case of carotid and subclavian stent placement plus vertebral artery revascularization) and thrombolysis was performed at variable times after stroke onset (range, 30 hours to 5 days). Four of the six patients had good basilar artery recanalization (Thrombolysis in Myocardial Infarction [TIMI] grade 0-1 before tissue plasminogen activator thrombolysis and TIMI grade 2 after procedure). Four of six patients had excellent immediate recovery and were discharged to an acute rehabilitation unit or their homes with improved neurologic symptoms and functional status. Two patients died: one patient presented with coma at outset with an NIHSS of 38, and the other patient probably had reocclusion of the basilar artery within 24 hours despite initial postprocedural improvement. CONCLUSION: We demonstrate that, in the setting of acute stroke, stent placement in combination with revascularization and thrombolysis is practical and allows quick access to a clot and simultaneously increases perfusion through collaterals during the thrombolytic process. In particular, basilar thrombolysis may be facilitated by proximal vertebral stent placement as concomitant atheromatous vertebrobasilar stenosis is common. PMID- 14561612 TI - Model of reversible cerebral ischemia in a monkey model. AB - We have developed a model of reversible cerebral ischemia in a high-level nonhuman primate. By using endovascular techniques, the posterior cerebral artery is permanently occluded with coils, and the ipsilateral middle cerebral artery is temporarily occluded with a balloon. The balloon can be deflated and/or removed to reestablish flow at precise time intervals. Functional imaging of the brain can be performed during occlusion and reperfusion, since the balloon can be deflated or removed in a scanner. PMID- 14561613 TI - Supplementary motor area activation in patients with frontal lobe tumors and arteriovenous malformations. AB - BACKGROUND AND PURPOSE: Some patients who undergo surgical resection of portions of the supplementary motor area (SMA) have severe postoperative motor and language deficits, whereas others have no deficits. We tested the hypothesis that in some patients with lesions affecting the SMA, the contralateral SMA exhibits some of the activation normally associated with the ipsilateral SMA. METHODS: Functional MR imaging studies in seven healthy volunteers and 19 patients with frontal lobe tumors or arteriovenous malformations were reviewed retrospectively. The hemisphere in which the SMA activation predominated was tabulated for right and left motor tasks. The relative hemispheric dominance in the SMA for the right and left motor tasks was compared in the healthy and patient groups and with the location of the lesion in the patient group. RESULTS: None of the control subjects performing a right hand motor task activated predominantly the right SMA. Fifty percent of the patients with lesions overlapping the left SMA performing the right motor task activated predominantly the right SMA. Fifty seven percent of control subjects performing the left hand motor task activated the left SMA predominantly. One hundred percent of patients with lesions overlapping the right frontal SMA performing the left motor task activated the left SMA predominantly. Differences between patients and controls were statistically significant. CONCLUSION: A lesion that contacts or overlaps the SMA is associated with an increased functional MR imaging response within the contralateral SMA. PMID- 14561614 TI - The anterior portion of the bilateral temporal lobes participates in music perception: a positron emission tomography study. AB - BACKGROUND AND PURPOSE: The mechanism underlying the perception of music has been the subject of study for many years. We investigated the role of the anterior portion of the temporal lobes in the perception of music in nonmusicians by use of positron emission tomography (PET). METHODS: We used the subtraction technique for PET to investigate the role of the anterior portion of the bilateral temporal lobes in music perception. Nonmusicians performed two kinds of musical tasks: harmony listening and soprano part listening. RESULTS: During the harmony listening task, the anterior portion of the temporal lobes, cingulate gyri, and cerebellum were bilaterally activated. During the soprano part-listening task, the bilateral superior parietal lobules, and the right precuneus were significantly activated. CONCLUSION: The anterior portion of the bilateral temporal lobes is vital in the discrimination of melodies and chords. Differences between activated brain regions exist between musicians and nonmusicians when listening to a particular vocal part of a musical phrase. PMID- 14561615 TI - Differential aging of the human striatum: longitudinal evidence. AB - BACKGROUND AND PURPOSE: Information about age-related changes in the striatum comes almost exclusively from cross-sectional studies. We examined age-related differences and longitudinal changes in the volume of the striatal nuclei, compared longitudinal measures of changes with their cross-sectional estimates, and addressed the question of differential aging of the basal ganglia. METHODS: We measured the volumes of the caudate nucleus (head), the putamen, and the globus pallidus on MR images of 53 healthy adults whose ages at baseline ranged between 20 and 77 years. The measurements were conducted twice, 5 years apart. RESULTS: On both measurement occasions, the volume of the neostriatal nuclei (caudate and putamen) but not of the globus pallidus correlated negatively with age. However, longitudinal comparison revealed significant declines in the neostriatal volumes and smaller (but statistically significant) shrinkage of the paleostriatum. In 5 years, the striatal components (caudate, putamen, and globus pallidus) shrunk by 1.21, 0.85, and 0.55 standard deviation units, respectively. The average annual shrinkage rate was 0.83% in the caudate nucleus, 0.73% in the putamen, and 0.51% in the globus pallidus. CONCLUSION: Although cross-sectional estimates suggested preservation of some striatal nuclei, longitudinal shrinkage of the whole striatum was evident even in a selected group of healthy adults. Moreover, the magnitude of observed longitudinal change was greater than predicted from cross-sectional studies. PMID- 14561616 TI - Diffusion tensor imaging of the hippocampal formation in temporal lobe epilepsy. AB - BACKGROUND AND PURPOSE: Diffusion tensor imaging (DTI) is a noninvasive technique that can be used to assess the integrity of cerebral tissue. The purpose of this study was to assess DTI measurements in the hippocampal formation (HF) and to investigate the role of DTI in lateralizing the seizure focus in temporal lobe epilepsy (TLE). METHODS: We evaluated 12 patients with unilateral TLE and 14 healthy subjects. We collected diffusion-weighted images along six different directions with a b value of 1000 s/mm(2), as well as an image acquired without diffusion weighting (b = 0 s/mm(2)). A 1.5-T imager was used to acquire 17 (3-mm) coronal sections covering the temporal lobes. We compared the mean diffusivity (trace D) and fractional anisotropy (FA) from symmetrical voxels by sampling the anterior HF bilaterally. We compared measurements with the EEG, high-resolution MR imaging, and clinical information. RESULTS: The patient group had significantly increased diffusivity and decreased FA in the HF ipsilateral to the seizure focus, as compared with values in the contralateral HF. When compared with healthy subjects, patients had significantly higher mean diffusivity in the ipsilateral HF; ipsilateral FA values were lower and did not reach statistical significance. Measurements in the contralateral HF did not show differences. Left right and absolute diffusivity indices lateralized the abnormal HF in eight and five of 12 patients, respectively. CONCLUSION: Abnormal DTI measurements and the epileptogenic HF are associated in unilateral TLE. This finding may reflect hippocampal sclerosis and may aid in presurgical evaluation. PMID- 14561617 TI - Relationship between the concentration of supplemental oxygen and signal intensity of CSF depicted by fluid-attenuated inversion recovery imaging. AB - BACKGROUND AND PURPOSE: Prior reports have described increased signal intensity (SI) of CSF on fluid-attenuated inversion recovery (FLAIR) images of anesthetized patients receiving 100% O(2). This appearance can simulate that of diseases. We evaluated the relationship between the concentration of inhaled O(2) and the development of increased SI of CSF on FLAIR images. METHODS: FLAIR was performed in 25 healthy volunteers breathing room air and 100% O(2) through a face mask for 5, 10, and 15 minutes. MR imaging, including FLAIR imaging, was performed in 52 patients with no potential meningeal abnormalities under general anesthesia: 21 received an equal mixture of N(2)O and O(2), and 31 received 100% O(2). The SI of CSF in volunteers and patients was graded in several locations by using a three point scale. RESULTS: SI of CSF significantly increased (P <.05) in various locations, in both volunteers and patients breathing 100% O(2), when compared with SI in the same volunteers breathing room air. Hyperintensity of CSF was not significantly different in volunteers receiving 100% O(2) through a face mask compared with anesthetized patients receiving 100% O(2) through a laryngeal airway or an endotracheal tube. No significant increase in SI occurred in patients receiving 50% O(2), when compared with the SI of volunteers breathing room air. CONCLUSION: Supplemental oxygen at 100% is a main cause of artifactual CSF hyperintensity on FLAIR images, regardless of the anesthetic drug used. This artifact does not develop when 50% O(2) is administered. PMID- 14561618 TI - Correlation of early dynamic CT perfusion imaging with whole-brain MR diffusion and perfusion imaging in acute hemispheric stroke. AB - BACKGROUND AND PURPOSE: Compared with MR imaging, dynamic CT perfusion imaging covers only a fraction of the whole brain. An important assumption is that CT perfusion abnormalities correlate with total ischemic volume. The purpose of our study was to measure the degree of correlation between abnormalities seen on CT perfusion scans and the volumes of abnormality seen on MR diffusion and perfusion images in patients with acute large-vessel stroke. METHODS: Fourteen patients with acute hemispheric stroke symptoms less than 12 hours in duration were studied with single-slice CT perfusion imaging and multislice MR diffusion and perfusion imaging. CT and MR perfusion studies were completed within 2.5 hours of one another (mean, 77 minutes) and were reviewed independently by two neuroradiologists. Hemodynamic parameters included cerebral blood flow (CBF), cerebral blood volume (CBV), and mean transit time (MTT). Extents of abnormality on images were compared by using Kendall correlation. RESULTS: Statistically significant correlation was found between CT-CBF and MR-CBF abnormalities (tau = 0.60, P =.003) and CT-MTT and MR-MTT abnormalities (tau = 0.65, P =.001). Correlation of CT-CBV with MR-CBV approached significance (tau = 0.39, P =.06). Extent of initial hyperintensity on diffusion-weighted images correlated best with extent of MR-CBV abnormality (tau = 0.69, P =.001), extent of MR-MTT abnormality (tau = 0.67, P =.002), and extent of CT-CBV abnormality (tau = 0.47, P =.02). CONCLUSION: Good correlation was seen between CT and MR for CBF and MTT abnormalities. It remains uncertain whether CT perfusion CBV abnormalities correspond well to whole-brain abnormalities. PMID- 14561619 TI - Hemodynamic changes after occlusion of the posterior superior sagittal sinus: an experimental PET study in cats. AB - BACKGROUND AND PURPOSE: Occlusion of the anterior third of superior sagittal sinus (SSS) is generally well tolerated because of sufficient collateral venous blood flow. In contrast, the pathophysiologic effects of occlusion of the SSS posterior to the rolandic vein remain controversial. We aimed to identify the specific hemodynamic effects of this subtype of SSS occlusion. METHODS: We ligated the SSS just behind rolandic vein and in the posterior part near the confluens sinus in three anesthetized cats. Regional cerebral blood flow (rCBF) was measured before and at 2 and 24 hours after the SSS occlusion. At around 48 hours, experimental settings were terminated with perfusion fixation with 4% paraformaldehyde solution. Hematoxylin-eosin histologic evaluation was performed. RESULTS: In all three cats with SSS occlusion, rCBF was reduced over the time period of measurement; this finding was observed in areas covering 5-20% of the brain in planes affected by the occlusion. The degree of rCBF reduction and the extension and severity of histologically proved venous infarction were correlated. CONCLUSION: To our knowledge, this is the first demonstration that occlusion of the SSS posterior to the rolandic vein is associated with a significant rCBF reduction to still-viable tissue in the related vascular territory at 24 hours after occlusion. We describe subacute venous infarction in an experimental occlusion of the SSS. Analogous to clinical conditions, occlusion of SSS alone without additional occlusion of bridging veins is adequate for producing a venous circulatory disturbance. PMID- 14561620 TI - MR imaging visualization of the cerebral microvasculature: a comparison of live and postmortem studies at 8 T. AB - SUMMARY: In this study, we compared 8-T gradient-echo MR images of the microvasculature in the live human brain with images of the unembalmed and embalmed postmortem brain. Small vessels were well visualized in the live brain and even better seen in the unembalmed postmortem brain, but they could not be visualized in the embalmed postmortem brain. These findings are important for direct comparisons of the microvasculature on 8-T MR images and on histologic sections. PMID- 14561621 TI - A rare congenital anastomosis between the vertebral artery and internal carotid artery. AB - A case of congenital anastomosis between the vertebral artery and internal carotid artery is presented. This rare anomaly was an incidental finding at cerebral angiography in a patient with a suspected ruptured cerebral aneurysm with subarachnoid hemorrhage. PMID- 14561622 TI - Neurotoxicity of intrathecal methotrexate: MR imaging findings. AB - We report a case of intrathecal methotrexate neurotoxicity manifesting as left arm weakness and aphasia. Diagnostic imaging showed restricted diffusion and fluid-attenuated inversion recovery imaging findings were normal at presentation. Three weeks later, diffusion abnormalities resolved, and T2-weighted studies showed increased signal intensity of prolonged T2 changes in areas of prior restricted diffusion. We attribute the clinical and radiographic findings to cytotoxic edema secondary to intrathecal methotrexate. PMID- 14561623 TI - American Society of Spine Radiology: the first decade. PMID- 14561624 TI - Kyphosis correction and height restoration effects of percutaneous vertebroplasty. AB - BACKGROUND AND PURPOSE: Percutaneous vertebroplasty is known for its pain relieving effect. Our purpose was to evaluate its effect on the kyphosis angle, wedge angle, and height of the fractured vertebral body. METHODS: We reviewed digital radiographs of 73 vertebral bodies in 53 patients before and after vertebroplasty. We measured the spinal kyphosis angle and the wedge angle of the fractured vertebral body. Ratios of the height of the anterior border, center, and posterior borders of the collapsed vertebra to the height at the posterior border of an adjacent normal vertebral body were measured. Gain from vertebroplasty and the restoration percentage (gain divided by loss) were calculated for each parameter. RESULTS: The kyphosis angle, wedge angle, anterior height, center height, and posterior height significantly improved after vertebroplasty. The mean reduction in the kyphosis angle was 4.3 degrees, and the wedge-angle reduction was 7.4 degrees. The mean wedge-angle reduction in fractured vertebral bodies containing gas was 10.2 degrees. Restoration percentages for the kyphosis angle and wedge angle were 19% and 44%, respectively. Gain in the height of the fractured vertebral bodies was 16.7% for the anterior border, 14% for the center, and 7% for the posterior border. Restoration percentages for the height of the vertebral body were 29% for the anterior border and 27% for the center. CONCLUSION: Vertebroplasty increases the height of the fractured vertebra and reduces the wedge and kyphosis angles. These effects are most remarkable in fractured vertebra containing gas. PMID- 14561625 TI - Slow-flow spinal epidural AVF with venous ectasias: two pediatric case reports. AB - We report the clinical and imaging findings in the cases of two children who initially presented with back pain related to epidural AVF in the cervicothoracic spine. Both lesions were of particular interest because of their exclusive epidural and paraspinal venous drainage and the presence of the prominent venous pouches in the epidural space. Angiography revealed that one was multifocal and of relatively slow flow. We think that these unusual features have important implications for treatment. PMID- 14561626 TI - MR imaging characteristics of pilomyxoid astrocytomas. AB - BACKGROUND AND PURPOSE: Pilomyxoid astrocytoma (PMA) is a recently described tumor that typically occurs in the chiasmatic-hypothalamic region in young children and has unique histopathologic and clinical characteristics. These tumors have been previously diagnosed as pilocytic astrocytoma (PA). PMA appears to have a higher rate of recurrence and CSF dissemination than typical PA. METHODS: We analyzed MR findings in four patients with PMA and compared them with those of typical chiasmatic-hypothalamic PA. RESULTS: MR findings of PMA were chiasmatic or hypothalamic enhancing solid tumor with hydrocephalus, highly homogeneous T2 signal intensity that extended into the deep white and gray matter, and CSF dissemination. CONCLUSION: Larger series are needed before the MR imaging findings of chiasmatic or hypothalamic enhancing solid tumor with hydrocephalus, highly homogeneous T2 signal intensity extending into the deep white and gray matter, and CSF dissemination can be used in the differential diagnosis of such tumors. PMID- 14561627 TI - High uptake on 11C-methionine positron emission tomographic scan of basal ganglia germinoma with cerebral hemiatrophy. AB - We herein describe a 12-year-old male patient with a germinoma of the basal ganglia who presented with progressive hemiparesis. MR imaging showed ipsilateral cerebral hemiatrophy predominantly in the left basal ganglia, whereas no mass or enhancement was depicted. Single photon emission CT revealed no significant uptake of thallium, whereas (11)C-methionine positron emission tomography showed clearly discernible uptake in the left putamen. Stereotactic biopsy, referencing the results of (11)C-methionine positron emission tomography, was performed, allowing histologic verification of germinoma to be established. (11)C-methionine positron emission tomography was the only technique that indicated the precise localization of the tumor in our patient and enabled biopsy-based final diagnosis of the basal ganglia germinoma without any overt mass formation. PMID- 14561628 TI - Early characteristics of Sturge-Weber syndrome shown by perfusion MR imaging and proton MR spectroscopic imaging. AB - We report the case of a 9-month-old boy with Sturge-Weber syndrome and new onset of seizure. Perfusion MR imaging showed early changes compatible with impaired venous drainage in the affected hemisphere, whereas proton MR spectroscopic imaging revealed a focal parietal area of elevated choline without significant alteration of N-acetylaspartate levels. The perfusion and subtle metabolic abnormalities are comparable with the extent of the overlying leptomeningeal enhancement, illustrating the early pathophysiological manifestation of this disease. PMID- 14561629 TI - Blue rubber-bleb nevus syndrome: report of a familial case with a dural arteriovenous fistula. AB - We report a case of a 45-year-old woman known to have a familial form of blue rubber-bleb nevus syndrome (BRBNS). The patient developed severe occipital headaches and bilateral retroauricular bruits. Cerebral angiography showed a large dural arteriovenous fistula in the torcular region. Central nervous system involvement in BRBNS is not often reported, and most cases of BRBNS are sporadic. PMID- 14561631 TI - Prenatal diagnosis of a vein of Galen aneurysmal malformation with fetal MR imaging study. PMID- 14561632 TI - The amphioxus Hairy family: differential fate after duplication. AB - Vertebrate Hairy genes are highly pleiotropic and have been implicated in numerous functions, such as somitogenesis, neurogenesis and endocrine tissue development. In order to gain insight into the timing of acquisition of these roles by the Hairy subfamily, we have cloned and studied the expression pattern of the Hairy gene(s) in amphioxus. The cephalochordate amphioxus is widely believed to be the living invertebrate more closely related to vertebrates, the genome of which has not undergone the massive gene duplications that took place early during vertebrate evolution. Surprisingly, we have isolated eight Hairy genes from the 'pre-duplicative' amphioxus genome. In situ hybridisation on amphioxus embryos showed that Hairy genes had experienced a process of subfunctionalisation that is predicted in the DDC model (for duplication degeneration-complementation). Only the summation of four out of the eight Amphi Hairy genes expression resembles the expression pattern of vertebrate Hairy genes, i.e. in the central nervous system, presomitic mesoderm, somites, notochord and gut. In addition, Amphi-Hairy genes expression suggest that amphioxus early somites are molecularly prefigured in an anteroposterior sequence in the dorsolateral wall of the archenteron, and the presence of a midbrain/hindbrain boundary. The expansion of the amphioxus Hairy subfamily request for caution when deducing the evolutionary history of a gene family in chordates based in the singularity of the amphioxus genome. Amphioxus may resemble the ancestor of the vertebrates, but it is not the ancestor, only its closest living relative, a privileged position that should not assume the freezing of its genome. PMID- 14561633 TI - A contradictory GLABRA3 allele helps define gene interactions controlling trichome development in Arabidopsis. AB - Previously characterized Arabidopsis gl3 mutants have trichomes that are smaller, less branched and undergo fewer rounds of endoreplication than wild-type trichomes. A new gl3 mutant, called gl3-sst, has oddly shaped trichomes that over expand during early development, undergo more endoreduplication and that have a striking nuclear morphology. The mutant nuclei consist of many interconnected lobes; however, only a single set of polytene-like chromosomes reside in the mutant nuclei. The predicted gl3-sst polypeptide has a Leu to Phe substitution (codon 78) within a region responsible for protein-protein interaction. Yeast interaction assays comparing GL3 with gl3-sst proteins show that the mutant protein interaction with GL1 and TTG1 is decreased by 75% and 50%, respectively, but there is no difference in its interaction with TRY. Furthermore, TRY has the ability to prevent the GL1 GL3 interaction and the GL1 gl3-sst interaction is even more sensitive to TRY. Analysis of plants expressing functional GFP-tagged versions of GL1, GL3 and TRY show that the proteins are localized in trichome nuclei. These results have been used to model trichome initiation in terms of protein interactions and threshold levels of activator complex. PMID- 14561634 TI - The role of buttonhead and Sp1 in the development of the ventral imaginal discs of Drosophila. AB - The related genes buttonhead (btd) and Drosophila Sp1 (the Drosophila homologue of the human SP1 gene) encode zinc-finger transcription factors known to play a developmental role in the formation of the Drosophila head segments and the mechanosensory larval organs. We report a novel function of btd and Sp1: they induce the formation and are required for the growth of the ventral imaginal discs. They act as activators of the headcase (hdc) and Distal-less (Dll) genes, which allocate the cells of the disc primordia. The requirement for btd and Sp1 persists during the development of ventral discs: inactivation by RNA interference results in a strong reduction of the size of legs and antennae. Ectopic expression of btd in the dorsal imaginal discs (eyes, wings and halteres) results in the formation of the corresponding ventral structures (antennae and legs). However, these structures are not patterned by the morphogenetic signals present in the dorsal discs; the cells expressing btd generate their own signalling system, including the establishment of a sharp boundary of engrailed expression, and the local activation of the wingless and decapentaplegic genes. Thus, the Btd product has the capacity to induce the activity of the entire genetic network necessary for ventral imaginal discs development. We propose that this property is a reflection of the initial function of the btd/Sp1 genes that consists of establishing the fate of the ventral disc primordia and determining their pattern and growth. PMID- 14561635 TI - Posterior patterning genes and the identification of a unique body region in the brine shrimp Artemia franciscana. AB - All arthropods share the same basic set of Hox genes, although the expression of these genes differs among divergent groups. In the brine shrimp Artemia franciscana, their expression is limited to the head, thoracic/trunk and genital segments, but is excluded from more posterior parts of the body which consist of six post-genital segments and the telson (bearing the anus). Nothing is currently known about the genes that specify the identity of these posterior structures. We examine the expression patterns of four candidate genes, Abdominal-B, caudal/Cdx, even-skipped/Evx and spalt, the homologues of which are known to play an important role in the specification of posterior structures in other animals. Abdominal-B is expressed in the genital segments of Artemia, but not in the post genital segments at any developmental stage. The expression of caudal, even skipped and spalt in the larval growth-zone suggests they may play a role in the generation of body segments (perhaps comparable with the role of gap and segmentation genes in insects), but not a direct role in defining the identity of post-genital segments. The expression of caudal at later stages suggests a role in the specification of anal structures. A PCR screen designed to isolate Hox genes expressed specifically in the posterior part of the body failed to identify any new Hox genes. We conclude that the post-genital segments of Artemia are not defined by any of the genes known to play a role in the specification of posterior segments in other arthropods. We argue that these segments constitute a unique body region that bears no obvious homology to previously characterised domains of Hox gene activity. PMID- 14561636 TI - Meiotic germ cells antagonize mesonephric cell migration and testis cord formation in mouse gonads. AB - The developmental fate of primordial germ cells in the mammalian gonad depends on their environment. In the XY gonad, Sry induces a cascade of molecular and cellular events leading to the organization of testis cords. Germ cells are sequestered inside testis cords by 12.5 dpc where they arrest in mitosis. If the testis pathway is not initiated, germ cells spontaneously enter meiosis by 13.5 dpc, and the gonad follows the ovarian fate. We have previously shown that some testis-specific events, such as mesonephric cell migration, can be experimentally induced into XX gonads prior to 12.5 dpc. However, after that time, XX gonads are resistant to the induction of cell migration. In current experiments, we provide evidence that this effect is dependent on XX germ cells rather than on XX somatic cells. We show that, although mesonephric cell migration cannot be induced into normal XX gonads at 14.5 dpc, it can be induced into XX gonads depleted of germ cells. We also show that when 14.5 dpc XX somatic cells are recombined with XY somatic cells, testis cord structures form normally; however, when XX germ cells are recombined with XY somatic cells, cord structures are disrupted. Sandwich culture experiments suggest that the inhibitory effect of XX germ cells is mediated through short-range interactions rather than through a long-range diffusible factor. The developmental stage at which XX germ cells show a disruptive effect on the male pathway is the stage at which meiosis is normally initiated, based on the immunodetection of meiotic markers. We suggest that at the stage when germ cells commit to meiosis, they reinforce ovarian fate by antagonizing the testis pathway. PMID- 14561637 TI - Central core disease: new findings in an old disease. PMID- 14561638 TI - Follicle stimulating hormone-induced DNA synthesis in the granulosa cells of hamster preantral follicles involves activation of cyclin-dependent kinase-4 rather than cyclin d2 synthesis. AB - Although cyclin D2 mRNA synthesis precedes gonadotropin-induced DNA synthesis in quiescent granulosa cells in culture, it is unclear whether a similar mechanism exists for the granulosa cells of growing preantral follicles in cyclic animals. The objective was to evaluate whether the synthesis of cyclin D2 protein was a prerequisite for FSH-induced DNA synthesis in the granulosa cells of intact preantral follicles of cyclic hamsters. Preantral follicles from cyclic hamsters were cultured in the presence or absence of FSH, and cell cycle parameters were examined. FSH stimulated cyclin-dependent kinase (CDK)-4 activity by 2 h and DNA synthesis by 4 h without altering the levels of cyclin D2 in the granulosa cells. The FSH effect was mimicked by epidermal growth factor administered in vivo. Although FSH increased the levels of cyclin D2 mRNA, it also stimulated the degradation of cyclin D2 as well as p27(Kip1) and p19(INK4) proteins. FSH activation of CDK4 was mediated by cAMP and ERK-1/2. In contrast to granulosa cells in intact follicles, FSH or cAMP significantly increased cyclin D2 protein levels in cultured granulosa cells but failed to induce DNA synthesis. Collectively, these data suggest that granulosa cells of preantral follicles, which are destined to enter the S phase during the estrous cycle, contain necessary amounts of cyclin D2 and other G1 phase components. FSH stimulation results in the formation and activation of the cyclin D2/CDK4 complex leading to DNA synthesis. This mechanism may be necessary for rapid movement of follicles from preantral to antral stages during the short duration of the murine estrous cycle. PMID- 14561639 TI - Anti-inflammatory and utero-relaxant effects in human myometrium of new generation phosphodiesterase 4 inhibitors. AB - The anti-inflammatory and utero-relaxant effects of two potent phosphodiesterase 4 (PDE4) inhibitors of the latest generation: cilomilast (one of the most advanced PDE4 inhibitors in clinical development, reportedly more selective for PDE4D) and compound A (which displays 12-fold greater selectivity toward PDE4B and/or PDE4A than toward PDE4D) were evaluated in human uterine smooth muscle. We first established that these compounds exhibit greater efficacy in inhibiting total cAMP-PDE activity in pregnant versus nonpregnant myometrium (E(max) = 78.0% +/- 3.6% and 80.3% +/- 2.2% in pregnant versus 57% +/- 4.7% and 70.5% +/- 5.9% in nonpregnant women for compound A and cilomilast, respectively; P < 0.05 for both compounds), confirming the prominent participation of PDE4 isoforms in cAMP hydrolysis in the near-term pregnant myometrium. Using pregnant myometrial explants, we have shown that both these drugs and also rolipram, the prototype PDE4 inhibitor, produce concentration-dependent inhibition of lipopolysaccharide (LPS) induced tumor necrosis factor alpha (TNFalpha) release with similar potency in each case (pD2 = 8.0 +/- 0.5, 7.9 +/- 0.2, and 7.6 +/- 0.2 for compound A, cilomilast, and rolipram, respectively). The maximum inhibition produced is 65%. Pretreatment with forskolin or 8-bromo-cAMP mimics the PDE4 inhibitor effect. Furthermore, compound A and cilomilast both produce concentration-dependent inhibition of the spontaneous contractions of myometrial strips and are more potent in pregnant than in nonpregnant myometrium (pD2 = 7.3 +/- 0.7 and 8.1 +/- 0.3 in pregnant versus 6.2 +/- 0.9 and 6.6 +/- 0.1 in nonpregnant myometrium for compound A and cilomilast, respectively; P < 0.05 for both compounds). This demonstrates that the PDE4 isoforms involved in the mechanism of contraction are different in the pregnant and nonpregnant myometrium. Our study highlights the importance of developing PDE4 inhibitors for the pharmacological management of infection-induced preterm labor. PMID- 14561640 TI - Identification of a member of a new RNase a family specifically secreted by epididymal caput epithelium. AB - In this study, we purified the first member of a new ribonuclease (RNase) A family from fluid of the proximal caput of the boar epididymis. This protein, named "Train A," is the most abundant compound secreted in the anterior part of the boar epididymis. After 2D electrophoresis, it is characterized by more than 10 isoforms ranging in size from 26 to 33 kDa and pI from 5 to 8.5. Several tryptic peptides were N-terminal sequenced, and an antiserum against one of these peptides was obtained. The protein was immunolocalized in the epididymal epithelium of the proximal caput, especially in the Golgi zone and the apical cytoplasm of the principal cells. In the lumen, spermatozoa were negative but droplets of reaction product were observed within the lumen. Full lengths of Train A cDNA were obtained from a lambdagt11 boar caput epididymis library and sequenced. The deduced protein is composed of 213 amino acids, including a 23 amino acid peptide signal and a potential N-glycosylation site. The mRNA of this protein has been retrieved and partially sequenced in the bull, horse, and ram, and homologous cDNA is found in databanks for the rat, mouse, and human. All the sequences are highly conserved between species. This protein and its mRNA are male-specific and exclusively expressed in the proximal caput of the epididymis, the only site where they have been found. Train A presents an RNase A family motif in its sequence. The RNase A family is a group of several short proteins (20-14 kDa) with greater and lesser degrees of ribonucleolytic activity and with supposed different roles in vivo. However, the presence of a long-conserved N terminal specific sequence and the absence of RNase catalytic site for Train A indicate that Train A protein is a member of a new family of RNase A. PMID- 14561641 TI - Immunity and beta-endorphin concentrations in hypothalamus and plasma in rats with steroid-induced polycystic ovaries: effect of low-frequency electroacupuncture. AB - The human endocrinological disorder polycystic ovary syndrome (PCOS) is a common cause of reproductive failure. Even though the cause of PCOS is unknown, hormone and immune disturbances as well as hyperactivity in the sympathetic nervous system are likely to be involved in the pathogenesis of the disease. The present study was undertaken to elucidate if rats with estradiol valerate (EV)-induced polycystic ovaries (PCO) have altered beta-endorphin concentrations in the hypothalamus and in plasma and if they have alterations in circulating immune cell populations and the activity. Repeated low-frequency (2 Hz) electroacupuncture (EA) treatments are known to modulate the release of beta endorphin, immune responses, and the activity in the autonomic nervous system. We therefore also investigated the effect of EA treatments on the beta-endorphin and the immune systems. Low-frequency EA was given 12 times, 25 min each, over 30 days starting 2-3 days after i.m. injection of EV. The beta-endorphin concentrations in the hypothalamus and in plasma as well as the frequencies of CD4+ T cells and CD8+ T cells were significantly lower in EV-injected control rats as compared to oil-injected control rats. Repeated EA treatments in EV injected rats significantly increased beta-endorphin concentrations in the hypothalamus. In conclusion, these findings show that both the beta endorphinergic and the immune system are significantly impaired in rats with steroid-induced PCO and that repeated EA treatments can restore some of these disturbances. PMID- 14561642 TI - Genetic changes in human fetuses from spontaneous abortion after in vitro fertilization detected by comparative genomic hybridization. AB - The in vitro fertilization (IVF) technique is becoming a very important approach for infertile disease therapy, but approximately 30% of pregnancies are spontaneously aborted in the first trimester. It is believed that chromosomal abnormality is the major reason for early spontaneous abortion. Although some reports have mentioned cytogenetic changes in spontaneously aborted embryos after IVF, little is known about the comprehensive cytogenetic alterations in these aborted embryos. Here we use the comparative genomic hybridization (CGH) technique to analyze the genetic alterations in 41 spontaneously aborted human specimens after IVF. In this study, 25 of 41 cases (61%) showed chromosomal changes. Among them, autosomes and sex chromosomes were involved in 16 and 11 cases, respectively. Several nonrandom chromosomal changes were identified, including loss of one sex chromosome (six cases) and gains of 22 (four cases), Y (four cases), 21 (three cases), 4 (two cases), and 13 (two cases). Our data support the opinion that chromosome abnormality is one of the major causes of early spontaneous abortion after IVF. The association between chromosome changes in these spontaneously aborted fetuses and maternal age, infertility patterns, infertility causes, and IVF patterns (routine IVF and other methods, including intracytoplasmic sperm injection, egg donation, and embryo donation) were also studied. No significant correlation was found. PMID- 14561643 TI - In vivo development of vitrified rat embryos: effects of timing and sites of transfer to recipient females. AB - In cryopreserved rat embryos, survival rates obtained in vitro are not always consistent with the rates obtained in vivo. To determine the optimal conditions for in vivo development to term, rat embryos at the 4-cell, 8-cell, and morula stages were vitrified in EFS40 by a one-step method and transferred into oviducts or uterine horns of recipients at various times during pseudopregnancy. Vitrified and fresh 4-cell embryos only developed after transfer into oviducts of asynchronous recipients on Days -1 to -2 of synchrony (i.e., at a point in pseudopregnancy 1-2 days earlier than the embryos). Approximately half the vitrified embryos transferred into oviducts on Day -1 developed to term, but only a minority of embryos, whether vitrified (10%-34%) or fresh (24%-33%), transferred at later times did so, suggesting that this may not be the most suitable stage for cryopreservation. Very few 8-cell embryos, either vitrified or fresh, developed when transferred into oviducts on Day 0 to -0.5. However, when transferred into uterine horns, high proportions of vitrified 8-cell embryos ( approximately 63%) developed to term in reasonably synchronous recipients (Day 0 to -0.5) but not in more asynchronous ones (6%; Day -1). A majority of vitrified morulae also developed to term (52%-68%) in a wider range of recipients (Days 0 to -1), the greatest success occurring in recipients on Day -0.5. Similar proportions of vitrified and fresh 4-cell embryos, 8-cell embryos, and morulae developed to term when appropriate synchronization existed between embryo and recipient. Thus, vitrification of preimplantation-stage rat embryos does not appear to impair their developmental potential in vivo. PMID- 14561644 TI - Pituitary glycoprotein hormone beta subunits in the Australian lungfish and estimation of the relative evolution rate of these subunits within vertebrates. AB - The beta subunits of the two pituitary gonadotropins LH and FSH and of thyroid stimulating hormone (TSH) were cloned from Australian lungfish (Neoceratodus forsteri) pituitary glands. These three glycoprotein hormone beta subunits possess the main characteristics common to their counterparts in other vertebrates. Taking advantage of the phylogenetic position of the lungfish, close to the root of tetrapods, a maximum parsimony tree was inferred from these new sequences and sequences from representatives of the diversity of vertebrates. The topology of the tree was imposed so that it reflected as closely as possible the real evolutionary history of the subunits. This tree was used to estimate the relative evolution rate of the three subunits in vertebrates. Cumulated amino acid substitutions from the basal subunit node (ancestral subunit sequence) to the species node were calculated and compared. It showed that a burst in evolutionary rate occurred for the LHbeta subunit in the tetrapod lineage sometime after the emergence of amphibians. The rate of evolution of the LHbeta subunit was particularly high throughout the radiation of mammals while FSH and TSHbeta subunits kept quite stable in this lineage. A burst in evolutionary rate was also observed for the FSHbeta subunit in the lineage leading to teleosts sometime after the emergence of chondrosteans and the dynamic of evolution was high throughout the radiation of teleosts. These results were consistent with data obtained from pairwise comparisons. PMID- 14561645 TI - Developmental potential of bovine androgenetic and parthenogenetic embryos: a comparative study. AB - In this study, we compared the developmental capacity of bovine haploid and diploid androgenetic and parthenogenetic embryos obtained by different methods. Androgenetic embryos were produced by piezo-intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF) of enucleated oocytes with or without subsequent pronuclear transfer from one haploid zygote to another. Parthenogenetic embryos were obtained by activation of matured oocytes by ionomycin combined with cycloheximide or 6-dimethylaminopurine (DMAP) treatment. Only few cleaved androgenetic haploid embryos were able to compact (2.7%) and to form blastocysts (1.8%), while significantly more haploid parthenogenotes underwent compaction (24-37%) and a minority developed to blastocysts at different rates, depending on the activation procedure (cycloheximide 3%, 6-DMAP 14.5%). By contrast, development to blastocyst of diploid androgenotes, cloned androgenetic embryos, and parthenogenotes (31%, 39%, and 43%, respectively) was similar to IVF control embryos (35%). Cell number on Day 7 was higher for IVF blastocysts and decreased in consecutive order in diploid androgenotes, diploid parthenogenotes, and haploid uniparental embryos. Following transfer of diploid androgenetic embryos, a pregnancy was established and maintained up to Day 28. PMID- 14561646 TI - Pituitary follistatin gene expression in female rats: evidence that inhibin regulates transcription. AB - Follistatin (FS), along with the members of the transforming growth factor beta family activin and inhibin, are important regulators of FSH secretion and messenger RNA production. While activin and inhibin appear to function as tonic modulators of FSH (stimulatory and inhibitory, respectively), dynamic changes in FS are noted through the estrous cycle and under varying physiological experimental paradigms. This suggests that FS is a major contributor to the precisely coordinated secretion of FSH that maintains reproductive function. The aim of this study was to investigate changes in FS, in particular the early (<12 h) rise observed after ovariectomy (OVX), and to determine whether these changes were as a consequence of variations in gene transcription rates. FS primary transcript (PT) and mRNA were found to increase 3-fold 12 h post-OVX, indicating increased gene transcription during this time period. Replacement with estradiol and/or blockade of GnRH had only modest effects on FS PT concentration. Inhibin immunoneutralization of intact rats resulted in a 3-fold increase in FS PT 12 h after administration of inhibin alpha antisera. Significant increases in FS mRNA at both 2 and 12 h also suggested that inhibin also may have effects on message stability. After administration of recombinant human inhibin A, there was a prompt decline in both FS PT and mRNA. These results indicate that inhibin is a major regulator of FS, both by transcriptional and nontranscriptional mechanisms. PMID- 14561647 TI - Effect of ubiquitin-proteasome pathway on mouse blastocyst implantation and expression of matrix metalloproteinases-2 and -9. AB - Previous studies have documented that ubiquitin-related proteins are present in human, baboon, rhesus monkey, cow, sheep, and mouse pregnant uteri, indicating that the ubiquitin-proteasome pathway (UPP) may be involved in the extensive uterine remodeling during mammalian early pregnancy, but there is still no direct evidence. A mouse intrauterine injection model was employed to study the direct effect of the UPP on mouse embryo implantation and its possible mechanisms. On Day 3 of pregnancy in each mouse, one of the uterine horns in each mouse was injected with different concentrations of lactacystin, a specific proteasome inhibitor, or anti-ubiquitin antibody, and the other side was used as a control. On days 5, 6, and 7, the number of implanted embryos was counted and the expression and gelatinolytic activities of matrix metalloproteinase-2 (MMP-2) and MMP-9 were studied. Results presented here illustrate that injection of lactacystin and anti-ubiquitin antibody significantly inhibited mouse embryo implantation. Further investigations by reverse transcription-polymerase chain reaction and gelatin zymography showed that MMP-2 and MMP-9 mRNA expression, as well as the gelatinolytic activity of MMP-9 in the lactacystin-treated uterine horn, significantly decreased, whereas the activity of MMP-2 was not significantly affected. The results obtained from this study, together with previous reports, suggest that the UPP is involved in mouse embryo implantation, and UPP's effect on embryo implantation is achieved at least in part by regulating MMP-2 and MMP-9 mRNA expression and the gelatinolytic activity of MMP 9. PMID- 14561648 TI - Activating transcription factor 4 is required for the differentiation of the lamina propria layer of the vas deferens. AB - Activating transcription factor 4 (ATF4/CREB2) is a member of the cyclic-AMP response element-binding (CREB) family. These proteins have been shown to regulate cell proliferation and differentiation in a broad number of tissues during embryo development. Here we report that male ATF4(-/-) mice are subfertile, despite the fact that they produce sufficient sperm and are able to fertilize wild-type eggs in vitro. An analysis of the ejaculatory ducts revealed abnormal constrictions in the lumen of the vas deferens. The lamina propria layer of the vas deferens was significantly thicker in the ATF4(-/-) mice and the cells that make up this layer were rounder and more abundant than in the ATF4(+/+) littermates. The change in the morphology of the lamina propria was associated with sexual maturation. A histologic analysis of the lamina propria revealed a reduction in the production of elastic fibers and interstitial cells of Cajal, as judged by the expression of neuron-specific enolase. These observations predict that ATF4 is required for the normal differentiation of the lamina propria layer of the vas deferens at sexual maturation. The morphology of the ATF4(-/-) lamina propria and the constriction of the lumen are consistent with an obstruction in the vas deferens contributing to the subfertility of the ATF4(-/-) males. PMID- 14561649 TI - DNA microarray analysis of region-specific gene expression in the mouse epididymis. AB - Microarray analysis was carried out to identify genes with enriched expression in the initial segment region of the mouse epididymis. A set of approximately 15 000 clones developed at the National Institutes for Aging and consisting of expressed sequence tags (ESTs) derived from pre- and peri-implantation embryos, Embryonic Day 12.5 female gonad/mesonephros, and newborn ovary were hybridized with probes generated against the initial segment (epididymal region 1) and the remainder of the epididymis (epididymal regions 2-5). The median values for the normalized ratios of region 1 to regions 2-5 from three independent experiments were averaged for each gene/EST using Genespring 5.0 software. The majority of clones showed a ratio of 1.0, suggesting they were expressed at similar levels in all epididymal regions. In addition, 123 clones exhibited 2-fold or higher expression in the initial segment, including Cres3, prostein, lipocalin 2, ALEX3, synaptotagmin-like 4, erm, and milk fat globule factor, whereas 216 clones, including elafin-like 1, lactotransferrin, Sin3B, zinc-finger protein 91, and membrane-type frizzled-related protein, showed 2-fold or higher expression in epididymal regions 2-5. Northern blot analyses of 12 clones predicted by microarray analysis to be either enriched in the initial segment (n = 8), enriched in epididymal regions 2-5 (n = 2), or similar in all regions (n = 2) were carried out. All clones exhibited the expected region-specific expression, thus confirming the microarray results. The studies presented here show a global survey of region-specific gene expression in the epididymis, identifying 15287 sequences, the majority of which have not previously been shown to be expressed in this organ. PMID- 14561650 TI - Effects of applying gamma-aminobutyric acid(B) drugs into the medial basal hypothalamus on basal luteinizing hormone concentrations and on luteinizing hormone surges in the female sheep. AB - Prior investigations have shown that localized infusion by microdialysis of gamma aminobutyric acid(B) (GABA(B)) agonists into the medial basal hypothalamus of male sheep rapidly increases GnRH and LH pulse amplitude. The objectives of these studies were to determine if infusion of GABA(B) agonists SKF 97541 or baclofen into the medial basal hypothalamus of female sheep would affect basal LH secretion and if infusion of a potent antagonist would alter expression of LH surges induced by injection of estrogen. Infusion of either SKF 97541 (10 or 40 microM) or baclofen (1 mM) into estrogen-treated ovariectomized ewes did not alter basal LH secretory patterns, whereas both drugs significantly elevated mean LH and LH pulse amplitude in ovariectomized ewes during the nonbreeding season. Infusion of the antagonist CGP 52432 (250 or 500 microM) did not affect expression of estrogen-induced LH surges in ovariectomized ewes. These observations support the concept that GABA(B) receptors in the medial basal hypothalamus regulate basal LH secretion but do not regulate the surge mode of LH secretion in the female sheep. PMID- 14561651 TI - Insulin-like growth factor-I and binding proteins 1, 2, and 3 in bovine nuclear transfer pregnancies. AB - In cloned pregnancies, placental deficiencies, including increased placentome size, reduced placentome number, and increased accumulation of allantoic fluid, have been associated with low cloning efficiency. To assess differences in paracrine and endocrine growth regulation in cloned versus normal bovine placentomes and pregnancies, we have examined the expression of insulin-like growth factor (IGF)-I and -II and their binding proteins (IGFBP)-1 through -3 in placentomes of artificially inseminated (AI), in vitro-produced (IVP), and nuclear transfer (NT) pregnancies at Days 50, 100, and 150 of gestation. Fetal, maternal, and binucleate cell counts in representative placentomes were performed on Days 50-150 of gestation in all three groups. Increased numbers of fetal, maternal, and binucleate cells were present in NT placentomes at all stages of gestation examined. Immunolocalization studies showed that spatial and temporal patterns of expression of IGFBP-2 and -3 were markedly altered in the placentomes of NT pregnancies compared to AI/IVP controls. Concentrations of IGF-I in fetal plasma, as determined by RIA, were significantly higher (P = 0.001) in NT pregnancies (mean +/- SEM, 30.3 +/- 2.3 ng/ml) compared with AI (19.1 +/- 5.5 ng/ml) or IVP (24.2 +/- 2.5 ng/ml) pregnancies on Day 150 of gestation. Allantoic fluid levels of IGFBP-1 were also increased in NT pregnancies. These findings suggest that endocrine and paracrine perturbations of the IGF axis may modulate placental dysfunction in NT pregnancies. Furthermore, increased cell numbers in NT placentomes likely have significant implications for fetomaternal communication and may contribute to the placental overgrowth observed in the NT placentomes. PMID- 14561652 TI - Gonadotropin-releasing hormone receptor gene expression during pubertal development of female rats. AB - Appropriate expression of the GnRH receptor (GnRH-R) in gonadotrophs is critical for GnRH signaling and hence for gonadotropin secretion and sexual development. In the present work, we have studied the ontogeny of the steady-state GnRH-R mRNA levels in pituitaries of female rats from Day 5 to Day 55, when sexual maturity is attained. Developmental changes of gonadotropin subunit (alpha, FSHbeta, and LHbeta) mRNA levels were also assessed. In addition, the role of the endogenous GnRH on the maturational changes of GnRH-R and gonadotropin subunit gene expression was investigated. Messenger RNA levels were determined by Northern blot analysis of total RNA from anterior pituitaries. Amounts of the most abundant (5.0 kilobase [kb]) GnRH-R mRNA increased slowly from Day 5 through the infantile period, to peak at Day 20 ( approximately 4-fold increase vs. Day 5). Thereafter the levels of the GnRH-R mRNA decline abruptly by Day 25 (75% decrease vs. Day 20) and then fell slightly until Day 35. Parallel changes were observed on the 4.5-kb transcript of the GnRH-R gene. Alpha subunit mRNA was easily detected at Day 5 and its levels increased quickly through the beginning of the infantile period to peak at Day 10 (3.2-fold increase vs. Day 5); then it decreased by 85% at Day 35. FSHbeta and LHbeta mRNA levels rose slowly until Days 15-20, a short time before GnRH-R. Thereafter, the levels of both mRNAs fell until Day 35 (90% decrease vs. Day 15 for FSHbeta and 50% decrease vs. Day 20 for LHbeta). To ascertain whether developmental activation of the GnRH-R and gonadotropin subunit gene expression is GnRH dependent, we have studied the effect of blocking the endogenous GnRH action by treating developing female rats with the specific GnRH antagonist cetrorelix (1.5 mg/kg body weight/wk, s.c.) through the infantile (Days 5-20) and the juvenile period (Days 20-35). Cetrorelix completely blocked the rise of levels of the two most abundant species, 5.0 kb and 4.5 kb, of GnRH-R mRNA during the infantile phase and dropped them to almost undetectable levels during the juvenile prepubertal period. Cetrorelix also abolished the developmental rise of gonadotropin beta subunit mRNAs during the two periods of the study. In contrast, alpha subunit gene expression tended to decrease, but not significantly, with cetrorelix treatment during the two periods. These data demonstrate that sexual maturation of female rats is advanced by an early and strong induction of GnRH-R and gonadotropin subunit gene expression during the infantile period, followed by weaker persistent activation during puberty. Developmental GnRH-R and gonadotropin beta subunit gene expression is almost entirely GnRH dependent, not only in the juvenile prepubertal stage but also during the infantile period. PMID- 14561653 TI - Expression of key prostaglandin synthases in equine endometrium during late diestrus and early pregnancy. AB - Luteolysis in domestic species is mediated by the release of luteolytic pulses of prostaglandin (PG) F(2alpha) by the uterus at the end of diestrus, which must be suppressed by the conceptus to permit maternal recognition of pregnancy. In many species, including the horse, both the conceptus and the endometrium also synthesize PGE(2), which may antagonize PGF(2alpha) by playing a luteotropic and/or antiluteolytic role. While the release of PGE(2) and PGF(2alpha) by the equine endometrium in late diestrus and early pregnancy has been previously studied, the underlying prostaglandin synthase gene regulatory mechanisms remain poorly defined. To resolve this issue, cyclooxygenase-2 (COX-2), microsomal PGE(2) synthase (PGES), and PGF(2alpha) synthase (PGFS) expression were examined in a series of endometrial biopsies obtained from cycling mares on Days 10, 13, and 15 postovulation, as well as from pregnant mares on Day 15. Quantification of COX-2 expression revealed significant (P < 0.01) increases in both mRNA and protein levels at Day 15 in cycling endometrium relative to other timepoints. Importantly, the level of COX-2 expression in Day 15 pregnant endometrium was found to be comparable with that observed in Day 10 and Day 13 cycling animals, suggesting that the presence of the conceptus blocks the induction of COX-2. Immunohistochemistry demonstrated that the induction of COX-2 expression on Day 15 occurs specifically in surface epithelial cells in cycling animals only. As equine PGFS had not been previously characterized, a 1380-base pair (bp) cDNA transcript was cloned by a combination of reverse transcription-PCR techniques and found to be highly homologous to bovine liver-type PGFS. The pattern of expression observed for the terminal PG synthases was distinct from that of COX 2, as PGES and PGFS mRNA and protein levels were found to be invariant throughout the timecourse and unaffected by pregnancy. Similar to COX-2, however, the PGES and PGFS proteins were found to localize mainly to the surface epithelium. Thus, this study describes for the first time the regulation and spatial distribution of COX-2, PGES, and PGFS expression in equine endometrium in late diestrus, with a marked induction of COX-2 but not of PGES and PGFS expression in uterine epithelial cells at Day 15. Furthermore, the presence of the conceptus was shown to block the induction of COX-2 expression at Day 15, suggesting an important mechanism by which it may suppress uterine PGF(2alpha) release and prevent luteolysis during early pregnancy. PMID- 14561654 TI - Differential expression of genes in the endometrium at implantation: upregulation of a novel member of the E2 class of ubiquitin-conjugating enzymes. AB - The process of embryo attachment and implantation is accompanied by dramatic cellular and functional changes in the endometrium, the control and mechanisms of which are not clearly understood. The cDNA cloning of differentially expressed genes, specifically at implantation sites in the rabbit endometrium, was used to identify genes controlling functional and remodeling changes. Tissue from the endometrium of Day 6(3/4) (preimplantation) and Day 8 (implantation initiation) pregnant rabbits was used to screen for differentially expressed genes by combined cDNA subtraction/suppressive hybridization. Twenty-nine differentially expressed genes were identified encoding protein modification enzymes, signaling proteins, structural proteins, and enzymes. One of these is a novel member of the E2 ubiquitin-conjugating enzyme family we have designated UBCi (i for implantation), which displayed dramatic nucleotide and deduced amino acid sequence conservation between rabbits, humans, and mice. In situ hybridization indicated UBCi expression exclusively in the luminal epithelium of the endometrium while glandular epithelium, trophoblast, and myometrium were negative. Expression was specific for epithelial cells at implantation sites and was not detected in non-implant-site endometrium. UBCi mRNA was detected in both the mesometrial and antimesometrial epithelial cells of the implantation sites, sites undergoing both differentiation and/or apoptosis. These results identify a group of differentially expressed genes in the endometrium including UBCi and provide new focal targets for studying processes controlling cellular remodeling during implantation. The important roles of ubiquitination in controlling the activities and turnover of key signaling proteins suggest potential roles in controlling critical aspects of implantation. PMID- 14561655 TI - Role of hydrogen peroxide in sperm capacitation and acrosome reaction. AB - The generation of reactive oxygen species (ROS) has been implicated in the regulation of sperm capacitation and acrosome reaction; however, the mechanisms underlying this regulation remain unclear. To examine the cellular processes involved, we studied the effect of different concentrations of hydrogen peroxide (H(2)O(2)) on protein tyrosine phosphorylation under various conditions. Treatment of spermatozoa with H(2)O(2) in medium without heparin caused a time- and dose-dependent increase in protein tyrosine phosphorylation of at least six proteins in which maximal effect was seen after 2 h of incubation with 50 microM H(2)O(2). At much higher concentrations of H(2)O(2) (0.5 mM), there is significant reduction in the phosphorylation level, and no protein tyrosine phosphorylation is observed at 5 mM H(2)O(2) after 4 h of incubation. Exogenous NADPH enhanced protein tyrosine phosphorylation similarly to H(2)O(2). These two agents, but not heparin, induced Ca(2+)-dependent tyrosine phosphorylation of an 80-kDa protein. Treatment with H(2)O(2) (50 microM) caused approximately a twofold increase in cAMP, which is comparable to the effect of bicarbonate, a known activator of soluble adenylyl cyclase in sperm. This report suggests that relatively low concentrations of H(2)O(2) are beneficial for sperm capacitation, but that too high a concentration inhibits this process. We also conclude that H(2)O(2) activates adenylyl cyclase to produce cAMP, leading to protein kinase A dependent protein tyrosine phosphorylation. PMID- 14561656 TI - Hormonal regulation and functional role of vascular endothelial growth factor a in the rat testis. AB - Vascular endothelial cell growth factor (VEGF-A) is synthesized in the testis but its role and regulation in this organ have not been examined. VEGF and its receptors (VEGF-R) were quantified using reverse transcription-polymerase chain reaction and Western blot. VEGF, VEGF-R1, and VEGF-R2 mRNAs and VEGF protein were increased after treatment with 50 IU hCG. Injection of 100 ng human recombinant VEGF 165 into the testis caused an increase in endothelial cell proliferation, but only a moderate increase in testicular interstitial fluid volume. In contrast with systemic hCG treatment, local VEGF injection did not increase the permeability to intravenously injected colloidal carbon particles. However, if VEGF was given locally in the testes of animals pretreated with hCG 4 or 8 h earlier, VEGF acted in synergy with hCG to increase vascular carbon leakage by forming interendothelial cell gaps. Testicular blood flow was unaffected by local VEGF 165 injection. Treatment with a specific VEGF-R2 tyrosine kinase inhibitor blocked the hCG-induced increase in endothelial cell proliferation but did not affect the hCG-induced accumulation of polymorphonuclear leukocytes in testicular blood vessels or the increase in the testicular interstitial space. The present study demonstrated that testicular VEGF secretion is increased by hormonal stimulation of Leydig cells and that VEGF, through effects mediated via VEGF-R2, regulates endothelial cell proliferation in the rat testis. VEGF does not appear to regulate testicular blood flow and it is not involved in inducing the hCG induced inflammation-like response in the testicular microvasculature. The permeability-increasing effect of VEGF is low in the testis under basal conditions but is apparently up-regulated by hCG treatment. PMID- 14561657 TI - Involvement of pro-inflammatory cytokines, mediators of inflammation, and basic fibroblast growth factor in prostaglandin F2alpha-induced luteolysis in bovine corpus luteum. AB - The process of luteolysis requires very subtly modulated coordination of different factors and regulation systems. Immune cells and cytokines were shown to be relevant for bovine luteolysis. The aim of this study was to investigate the detailed pattern of mRNA expression of the pro-inflammatory cytokines tumor necrosis factor alpha (TNFalpha), TNF receptor type 1 (TNF-R1), interleukin 1beta (IL-1beta), and interferon gamma (IFNgamma), and of the inducible nitric oxide synthase (iNOS) and the basic fibroblast growth factor (FGF-2) during prostaglandin (PG) F(2alpha)-induced luteolysis in the bovine corpus luteum (CL). In addition, the mRNA expression for the LH-receptor (LH-R) and the steroidogenic enzyme p450scc was determined. Cows in the midluteal phase (Days 8-12) were injected with the PGF(2alpha) analogue cloprostenol, and CL were collected by transvaginal ovariectomy before and 2, 4, 12, 48, and 64 h after PGF(2alpha) injection. Conventional and real-time reverse transcription polymerase chain reaction RT-PCR (LightCycler) using SYBR Green I detection were employed to determine the mRNA expression for the investigated factors. All cytokines were significantly up-regulated during induced luteolysis. LH-R and p450scc mRNA were down-regulated (P < 0.05) during structural luteolysis (after 12 h), and p450scc in addition at 2 h after PGF(2alpha) (P < 0.05). FGF-2 expression increased (P < 0.001) during functional luteolysis (until 12 h after PGF(2alpha)) and diminished thereafter. The mRNA expression for iNOS decreased (P < 0.05) after induction of luteolysis. In conclusion, cytokines may be involved not only in structural but also in functional luteolysis and the deprivation of luteal survival factors, leading to a situation where apoptosis can occur. FGF-2 may participate in the suppression of cytokine-induced iNOS mRNA expression and in the prevention of an inflammatory reaction in the surrounding tissues. PMID- 14561658 TI - Age-related changes in the ultrastructure of the resting follicle pool in human ovaries. AB - Age-related decline of fertility in women is the result of the decline in both quantity and quality of the resting ovarian follicle pool. The aim of the present study was to determine whether the decline of follicle quality with age is reflected by ultrastructural changes in the resting follicle pool. Ovarian biopsy specimens were obtained by laparoscopy from seven healthy women aged 25-32 yr (young group) and from 11 healthy women aged 38-45 yr (advanced-age group). A total of 182 resting follicles from the young group were compared with 81 resting follicles from the advanced-age group for signs of age-related changes by transmission-electron microscopy. The ooplasmic fraction of vacuoles was increased (P = 0.02), and the fraction of mitochondria decreased (P = 0.005), in the advanced-age group. Also, the density of the mitochondrial matrix (P < 0.001) and the frequency of dilated smooth endoplasmic reticulum (SER; P = 0.001) and Golgi complex (P = 0.02) were increased with age. The frequencies of ruptured mitochondrial membranes (P = 0.001) and dilated SER (P = 0.003) were increased with age in the granulosa cells. Overall follicle-quality scores, which should reflect atretic changes, were not different for the young and advanced-age groups. In conclusion, in resting follicles, the morphological changes with age are different from the changes seen in quality decline by atresia. The morphological changes with age specifically involved the mitochondria, the SER, and the Golgi complex, and they may be the cause of atresia on initiation of follicular growth because of the substantial increase in metabolic requirements. PMID- 14561659 TI - Thrombospondin 2 deficiency in pregnant mice results in premature softening of the uterine cervix. AB - The gradual disorganization of collagen fibers in the stromal connective tissue of the uterine cervix is characteristic of progressive cervical softening during pregnancy. A lack of thrombospondin (TSP) 2 has been shown to be associated with altered collagen fibril morphology of connective-tissue-rich organs such as skin and tendon. The goal of this study was to determine the role of TSP2 in cervical softening by studying a TSP2-null mouse line. Creep testing showed that, in the nonpregnant animal and on Day 10 of pregnancy, there was no difference between the cervical extensibility of the wild-type and the TSP2-deficient mice. However, by Day 14 of pregnancy, the TSP2-null mice showed 4.5-fold increase in cervical extensibility, and by Day 18, a 6.1-fold increase, when compared with wild-type mice. A further indicator of compromised cervical integrity was that, on Days 14 and 18 of pregnancy, the cervix of TSP2-null mice broke rapidly under standard loading conditions that did not break the cervix of wild-type mice. Western blotting showed that TSP2 was expressed in the cervix of mice on Days 14 and 18 of pregnancy but not on Day 10 or in the nonpregnant animal. As determined by immunohistochemistry, the amount of matrix metalloproteinase 2 (MMP2) in the cervix of TSP2-null mice increased 11-fold on Day 14 of pregnancy and 19-fold on Day 18. Thus, TSP2-null mice provide an animal model to assist in the understanding of the molecular basis of spontaneous, premature softening of the uterine cervix. PMID- 14561660 TI - Analysis of historical data suggests long-lasting protective effects of smallpox vaccination. AB - More than half of the US population has received the smallpox vaccine, but it is unknown what fraction is still protected against infection and disease. Residual protection and age-dependent case-fatality ratios have therefore been widely neglected in the current bioterrorism debate. The author analyzed 1902-1903 data from Liverpool, United Kingdom, and from reintroductions of the disease to Europe in 1950-1971 to estimate to what degree vaccinated cases were protected against developing severe or fatal disease and how quickly this protection waned over time. Protection against severe and fatal disease was lost at the rate of 1.41% per year, corresponding to a half-life of 49.2 years (95% confidence interval: 42.0, 57.3), and protection against fatal disease alone declined 0.363% per year. Thus, even 70 years after primary vaccination, 77.6% of cases were still protected (95% confidence interval: 66.6, 85.4). Protection against severe disease should therefore extend for many decades after a single vaccination, and protection against death from smallpox may even be lifelong for the majority of vaccinees. This protection should greatly reduce the number of severe and fatal cases of disease expected in a bioterrorist attack, but residual protection may also increase the risk that some previously vaccinated cases who develop mild disease may remain unrecognized longer, while moving around freely and disseminating the infection. PMID- 14561661 TI - Birth weight as a risk factor for childhood leukemia: a meta-analysis of 18 epidemiologic studies. AB - Evidence has emerged that childhood leukemia is initiated in utero. High birth weight is one of the few birth-related factors that has been associated with childhood leukemia, albeit not consistently. The authors conducted a meta analysis of studies of the association between birth weight and childhood leukemia risk. Study-specific odds ratios for leukemia were calculated, using a cutoff at 4,000 g of birth weight. The authors also evaluated whether the association between birth weight and leukemia followed a log-linear dose-response like pattern. They calculated summary estimates using weighted averages of study specific odds ratios from dichotomous and trend analyses. Eighteen studies (published between 1962 and 2002) were included, encompassing 10,282 children with leukemia. Children weighing 4,000 g or more at birth were at higher risk of acute lymphoblastic leukemia than children weighing less (odds ratio (OR) = 1.26, 95% confidence interval (CI): 1.17, 1.37). Furthermore, data were consistent with a dose-response-like effect (OR = 1.14/1,000-g birth weight increase, 95% CI: 1.08, 1.20). Studies of acute myeloid leukemia indicated a similar increase in risk for children weighing 4,000 g or more at birth (OR = 1.27, 95% CI: 0.73, 2.20) and a dose-response-like effect (OR = 1.29/1,000 g, 95% CI: 0.80, 2.06), but results varied across studies. Our findings support a relation between birth weight and childhood acute lymphoblastic leukemia risk and emphasize the need for additional studies of the biologic mechanisms underlying this association. PMID- 14561662 TI - Contribution of socioeconomic status to the association between hostility and cardiovascular risk behaviors: a prospective cohort study. AB - The authors examined the contribution of childhood and early adulthood socioeconomic status (SES) to the association between adulthood cynical hostility and cardiovascular risk behaviors. Participants from the population-based, prospective Cardiovascular Risk in Young Finns Study were 531 males and 688 females, aged 12-21 years at the baseline in 1983 and 21-30 years at the follow up in 1992. Cardiovascular risk behaviors comprised the number of cigarettes smoked per day, physical inactivity, the type of fat used in the diet, and the frequency of alcohol consumption. The general linear models showed socioeconomic variation in cynical hostility, butter use in the diet, and smoking. In regression analyses, hostility was positively associated with smoking in men and women (beta coefficients = 0.16 and 0.09; p values = 0.000 and 0.019, respectively) and with frequency of alcohol use (beta coefficients = 0.10 and 0.03; p values = 0.024 and 0.03, respectively). Adding parents' and participants' SES to the model marginally attenuated these associations. The authors conclude that the association of cynical hostility with smoking and alcohol use seems to be independent of intergenerational social mobility and childhood and adulthood SES. PMID- 14561663 TI - Invited commentary: socioeconomic status, hostility, and health behaviors--does it matter which comes first? PMID- 14561664 TI - Genetic variation of infant reduced folate carrier (A80G) and risk of orofacial and conotruncal heart defects. AB - How folate reduces the risks of congenital anomalies is unknown. The authors focused on a gene involved in folate transport-reduced folate carrier-1 gene (RFC1). Using data from a California case-control study (1987-1989 births), the authors investigated whether the risks of orofacial clefts or conotruncal heart defects were influenced by a polymorphism of infant RFC1 or by an interaction between the RFC1 gene and maternal periconceptional use of vitamins containing folic acid. A total of 305 liveborn infants with cleft lip with or without cleft palate, 123 with cleft palate, 163 with conotruncal heart defects, and 364 nonmalformed controls were genotyped. Odds ratios of 1.6 (95% confidence interval: 0.9, 2.8) for the G80/G80 genotype and of 2.3 (95% confidence interval: 1.3, 3.9) for the G80/A80 genotype were observed relative to the A80/A80 genotype for conotruncal defects. Among mothers who did not use vitamins, the risk of conotruncal defects was 2.1 (95% confidence interval: 0.7, 5.9) for infants with genotype G80/G80 compared with those with the A80/A80 genotype. Among mothers who did use vitamins, the risk was 1.3 (95% confidence interval: 0.7, 2.7). Substantially elevated risks for either cleft group were not observed irrespective of genotype and use/nonuse of vitamins. Thus, this study found modest evidence for a gene-nutrient interaction between infant RFC1 genotype and periconceptional intake of vitamins on the risk of conotruncal defects. PMID- 14561665 TI - Severity factors for truck drivers' injuries. AB - A study was carried out in 1995-1999 to assess severity factors for truck drivers' crashes. The authors used data from the trauma registry of road crash victims of the Rhone region, France. Several descriptive characteristics of the victims (age, place of residence) and their crashes (place, time, antagonist, seatbelt wearing) were analyzed. The injuries of 300 male truck drivers were described by body region, and their severity was measured by using the injury severity score comparing these drivers with 9,488 male car drivers (age: 18-67 years). Truck drivers were more seriously injured than car drivers; the odds ratio was 1.87 (95% confidence interval: 1.33, 2.63) for having an injury severity score of 9 or more. Limb and abdominal lesions were more frequent and more serious among truck drivers. The lack of seatbelt wearing by truck drivers was one of the factors that explained the particular severity of their injuries; the odds ratio, adjusted for seatbelt wearing, for truck drivers to be seriously injured was 1.04 (95% confidence interval: 0.73, 1.47) compared with car drivers. When all of the severity factors were taken into account, the risk was even lower, but not significantly so. PMID- 14561666 TI - Joint association of alcohol and folate intake with risk of major chronic disease in women. AB - Alcohol interferes with folate metabolism and has opposing effects on the risks of cardiovascular disease and cancer. The authors examined the joint association of alcohol and folate intake with risk of major chronic disease, defined as fatal or nonfatal cardiovascular disease or cancer, or other nontraumatic death. This study included 83,929 women aged 34-59 years with no previous history of cardiovascular disease or cancer who provided dietary data in 1980. During 16 years of follow-up, the authors documented 10,666 new cases of major chronic disease. Overall, heavy drinkers (>30 g/day) with a lower total folate intake (<180 micro g/day) had the highest risk; in comparison with abstainers with a folate intake of 400-599 micro g/day, the multivariate relative risk was 1.36 (95% confidence interval: 1.10, 1.70). However, the increased risk of major chronic disease associated with heavy drinking was largely diminished among women with a higher folate intake (p for interaction = 0.02). The positive association between heavy alcohol/low folate intake and risk of major chronic disease was most apparent among women younger than age 60 years. Adequate folate intake may be important in the primary prevention of overall major chronic disease in women, especially among younger women consuming more than two alcoholic drinks per day. PMID- 14561667 TI - Habitual tea consumption and risk of osteoporosis: a prospective study in the women's health initiative observational cohort. AB - The purpose of this study was to prospectively investigate associations of habitual drinking of regular tea with bone mineral density and fracture risk. Study participants were a multiethnic postmenopausal cohort (n = 91,465) from the nationwide Women's Health Initiative Observational Study. These women were recruited in the United States and aged 50-79 years at the time of enrollment (1994-1998). The average follow-up time was 4.1 years. Habitual consumption of regular tea was assessed with a structured questionnaire at baseline. Clinical fractures during the follow-up were reported in questionnaires, and hip fractures were further confirmed by reviewing medical records. Bone mineral density measurements were conducted among a subgroup of women (n = 4,979) at three Women's Health Initiative bone mineral density centers using dual-energy x-ray absorptiometry. Multivariate analyses suggested a positive trend of increased total body bone mineral density with tea drinking (p < 0.05). However, results from the Cox proportional hazard models did not show any significant association between tea drinking and the risk of fractures at the hip and forearm/wrist. In conclusion, the results from this study indicate that the effect of habitual tea drinking on bone density is small and does not significantly alter the risk of fractures among the US postmenopausal population. PMID- 14561668 TI - Timing of menopause and patterns of menstrual bleeding. AB - Age at menopause is associated with a variety of health outcomes. Menstrual histories, both as markers of physiologic function and through their potential association with age at menopause, have also been investigated for their links to health outcomes. This study used data from a cohort of women from the United States who provided prospectively recorded data on their menstrual cycles for many years. Dr. Alan Treloar (University of Minnesota) originally recruited the women in the 1930s; the authors used data reported by these women from 1930 through 1977. They identified nuanced characteristics of menstrual histories that were strongly predictive of the onset of menopause, focusing on measures of central tendency (the mean), variability (standard deviation), and serial irregularity (approximate entropy), the last of which quantifies a continuum that ranges from totally ordered to completely random. They controlled for other characteristics known to affect age at menopause, including use of exogenous hormones, number of births, and extent of breastfeeding. Although cycle length and variability increased with the approach of menopause, the authors found that serial irregularity decreased and was a strong predictor of its onset. This finding constitutes an important piece of information not attainable with conventional statistical summaries of menstrual histories. PMID- 14561669 TI - Differences in body shape representations among young adults from a biracial (Black-White), semirural community: the Bogalusa Heart Study. AB - In this study, the authors examined body image perception and body mass index (weight (kg)/height (m)(2)) among race-gender groups in a biracial (Black-White) population of young adults in Bogalusa, Louisiana. A mail-out survey was completed in 1994 by 3,698 (65%) participants aged 18.5-35 years in the Bogalusa Heart Study (mean age = 27.6 years). As part of the survey, body image perception was determined in terms of body shape representations from a figure rating scale. A body image discrepancy score was calculated from the difference between z standardized values of body image perception and body mass index. A stepwise proportional odds model including the covariates income, employment, education, and physical activity was used to identify factors influencing lower perception of body shape. Mean body mass index was highest among Black females (p < 0.001). The odds of having a lower perception of body shape (vs. body mass index) were 1.72 times higher in Blacks (p < 0.001), 0.80 times lower in persons who were currently employed (p < 0.001), and 0.86 times lower in persons with a higher education (p = 0.032). Gender, income, and physical activity were not found to be significant predictors of body image perception (p > 0.05). The authors conclude that significant differences exist within racial groups concerning body image perception in relation to overweight status among young adults. This has implications for prevention and education programs. PMID- 14561670 TI - Use of electric bedding devices and risk of breast cancer in African-American women. AB - In this case-control study, the authors aimed to examine whether use of an electric bedding device increased breast cancer risk in African-American women. Cases were 304 African-American patients diagnosed with breast cancer during 1995 1998 who were aged 20-64 years and lived in one of three Tennessee counties. Controls were 305 African-American women without breast cancer who were selected through random digit dialing and frequency-matched to cases by age and county. Information on the use of an electric blanket or heated water bed and other risk factors was collected through telephone interviews. Breast cancer risk associated with use of an electric bedding device increased with the number of years of use, the number of seasons of use, and the length of time of use during sleep. When women who used an electric bedding device for more than 6 months per year (and therefore were more likely to have used a heated water bed, which generates lower magnetic fields) were excluded, the corresponding dose-response relations were more striking. Similar trends in dose response were shown in both premenopausal and postmenopausal women and for both estrogen receptor-positive and estrogen receptor-negative tumors. The use of electric bedding devices may increase breast cancer risk in African-American women aged 20-64 years. Such an association might not vary substantially by menopausal status or estrogen receptor status. PMID- 14561671 TI - Prolonged detection of indigenous wild polioviruses in sewage from communities in Egypt. AB - Environmental surveillance for polioviruses has been implemented in Egypt. This paper reports on a study in which 130 sewage samples were collected between January 2001 and December 2001 from eight provinces of Egypt. Samples were analyzed by virus isolation in L20B and RD cell cultures, and wild polioviruses were characterized by sequencing of the VP1 protein coding region. Wild type 1 polioviruses were detected in 57% of the sewage samples and 91% of the study sites, only two of which reported paralytic poliomyelitis cases in 2001. Three genetic lineages of a single indigenous type 1 poliovirus genotype were detectable in sewage, and only one lineage was also detected through surveillance for acute flaccid paralysis. Wild polioviruses persisted in the environment despite implementation of oral poliovirus vaccine immunization campaigns. Continued analysis of sewage samples, critical evaluation of immunization coverage, and performance of surveillance for acute flaccid paralysis are proposed as follow-up activities. PMID- 14561672 TI - Determinants of geographic variation in Helicobacter pylori infection among children on the US-Mexico border. AB - Rates of Helicobacter pylori infection are traditionally higher in developing countries than in developed countries, but the specific reasons for these differences are not fully clear. While chronic diseases resulting from H. pylori are generally of adult onset, chronic infection usually begins in childhood. In this cross-sectional study (1998-2000), the authors compared prevalences of H. pylori infection among children under age 6 years on both sides of the Rio Grande. Participants included 264 children of women from low-income families who were receiving services at health clinics in Juarez, Mexico, or El Paso, Texas, from April 1998 through October 2000. Data were collected through personal interviews and serologic testing for H. pylori antibodies. The crude odds ratio for H. pylori prevalence among Mexican children as compared with US children was 3.94 (95% confidence interval: 1.72, 9.06). After adjustment for covariates, the odds ratio decreased to 1.70 (95% confidence interval: 0.64, 4.52). The adjustments that produced the greatest reduction in the odds ratio for location were those for household crowding and maternal education. This study identified specific factors that may explain geographic variation in H. pylori prevalence among children. PMID- 14561674 TI - Re: "Abortion,changed paternity, and risk of preeclampsia in nulliparous women". PMID- 14561675 TI - The vascular contribution in the pathogenesis of inflammatory bowel disease. PMID- 14561676 TI - Acute adenosinergic cardioprotection in ischemic-reperfused hearts. AB - Cells of the cardiovascular system generate and release purine nucleoside adenosine in increasing quantities when constituent cells are "stressed" or subjected to injurious stimuli. This increased adenosine can interact with surface receptors in myocardial, vascular, fibroblast, and inflammatory cells to modulate cellular function and phenotype. Additionally, adenosine is rapidly reincorporated back into 5'-AMP to maintain the adenine nucleotide pool. Via these receptor-dependent and independent (metabolic) paths, adenosine can substantially modify the acute response to ischemic insult, in addition to generating a more sustained ischemia-tolerant phenotype (preconditioning). However, the molecular basis for acute adenosinergic cardioprotection remains incompletely understood and may well differ from more widely studied preconditioning. Here we review current knowledge and some controversies regarding acute cardioprotection via adenosine and adenosine receptor activation. PMID- 14561677 TI - Increased calcium intake reduces plasma cholesterol and improves vasorelaxation in experimental renal failure. AB - Chronic renal failure (CRF) is associated with abnormal lipid metabolism and high prevalence of vascular complications. Calcium salts are commonly used in CRF as phosphate binders. Increased calcium intake may also lower plasma cholesterol and beneficially influence vascular tone. Therefore, we investigated the influence of increasing dietary calcium from 0.3% to 3.0% for 8 wk after 5/6 nephrectomy (NTX) on plasma cholesterol and mesenteric resistance vessel tone in male Sprague Dawley rats. The groups were Sham, Sham-Calcium, NTX, and NTX-Calcium (n = 10 11). Blood pressure was modestly elevated after NTX, whereas the plasma creatinine, urea nitrogen, phosphate, and parathyroid hormone levels were clearly increased. The high-calcium diet suppressed plasma phosphate and parathyroid hormone but was without effect on blood pressure. The NTX resulted in 1.6-fold elevation in plasma total cholesterol and 40% reduction in high density-to-low density lipoprotein ratio (HDL/LDL). However, the lipid profile in NTX rats on the high-calcium diet did not differ from sham-operated controls. The endothelium mediated relaxations induced by acetylcholine were impaired in NTX rats, whereas the response was normalized by a high-calcium diet. No differences in vasorelaxations by the endothelium-independent vasodilator nitroprusside were detected. In conclusion, improved vasorelaxation after a high-calcium diet could be due to reduced plasma total cholesterol and ameliorated HDL/LDL ratio, although decreased plasma phosphate and parathyroid hormone may also play a significant role in the vascular effects of increased calcium intake. PMID- 14561678 TI - RV instantaneous intraventricular diastolic pressure and velocity distributions in normal and volume overload awake dog disease models. AB - Intraventricular diastolic right ventricular (RV) flow field dynamics were studied by functional imaging using three-dimensional (3D) real-time echocardiography with sonomicrometry and computational fluid dynamics in seven awake dogs at control with normal wall motion (NWM) and RV volume overload with diastolic paradoxical septal motion. Burgeoning flow cross section between inflow anulus and chamber walls induces a convective pressure rise, which represents a "convective deceleration load" (CDL). High spatiotemporal resolution dynamic pressure and velocity distributions of the intraventricular RV flow field revealed time-dependent, subtle interactions between intraventricular local acceleration and convective pressure gradients. During the E-wave upstroke, the total pressure gradient along intraventricular flow is the algebraic sum of a pressure decrease contributed by local acceleration and a pressure rise contributed by a convective deceleration that partially counterbalances the local acceleration gradient. This underlies the smallness of early diastolic intraventricular gradients. At peak volumetric inflow, local acceleration vanishes and the total adverse intraventricular gradient is convective. During the E-wave downstroke, the strongly adverse gradient embodies the streamwise pressure augmentations from both local and convective decelerations. It induces flow separation and large-scale vortical motions, stronger in NWM. Their dynamic corollaries on intraventricular pressure and velocity distributions were ascertained. In the NWM pattern, the strong ring-like vortex surrounding the central core encroaches on the area available for flow toward the apex. This results in higher linear velocities later in the downstroke of the E wave than at peak inflow rate. The augmentation of CDL by ventriculoannular disproportion may contribute to E wave and E-to-A ratio depression with chamber dilatation. PMID- 14561679 TI - Shear modulus of porcine coronary artery: contributions of media and adventitia. AB - The epicardial coronary arteries experience significant torsion in the axial direction due to changes in the shape of the heart during the cardiac cycle. The objective of this study was to determine the torsional mechanical properties of the coronary arteries under various circumferential and longitudinal loadings. The coronary artery was treated as a two-layer composite vessel consisting of intima-medial and adventitial layers, and the shear modulus of each layer was determined. Eight porcine hearts were obtained at a local abattoir, and their right coronary and left anterior descending arteries were isolated and tested in vitro with a triaxial torsion machine (inflation, longitudinal stretch, and circumferential twist). After the intact vessel was tested, the adventitia was dissected away, leaving an intact media that was then tested under identical triaxial loading conditions. We proposed a biomechanical analysis to compute the shear modulus of the adventitia from the measured shear moduli of the intact vessel and the media. To validate our predictions, we used four additional hearts in which the shear modulus of the adventitia was measured after dissection of media. Our results show that the shear modulus does not depend on the shear stress or strain but varies linearly with circumferential and longitudinal stresses and in a nonlinear way with the corresponding strains. Furthermore, we found that the shear modulus of the adventitia is larger than that of the intact vessel, which is larger than the vessel media. These results may have important implications for baroreceptor sensitivity, circulation of the vasa vasorum, and coronary dissection. PMID- 14561680 TI - Microcirculatory changes during chronic adaptation to hypoxia. AB - Microcirculatory changes in the window chamber preparation in Syrian golden hamsters, secondary to chronic hypoxia adaptation, are presented herein. Adaptation was attained by keeping animals in a 10% oxygen environment for 1 wk and 5% the following week. The following groups were studied: group 1, adapted to chronic hypoxia and kept in a 5% oxygen environment throughout the experiment; group 2, adapted to chronic hypoxia and kept in a 21% oxygen environment 24 h before and during the experiment; and group 3, control. Adaptation caused venule enlargement and hematocrit increase (68.6 +/- 2.44 in group 1, 70 +/- 2.66 in group 2, and 43.27 +/- 2.30 in group 3; P < 0.05). Whereas heart rate decreased in adapted animals, blood pressure remained constant. Group 1 presented alkalosis, hypocapnia, and hypoxemia. The adapted groups had decreased blood flow velocity in arterioles and veins. We found no difference in microvasculature oxygen tension between groups 2 and 3; however, the number of capillaries with flow was markedly reduced in group 1 but significantly increased in group 2. Our findings suggest that, as an adaptation to hypoxia, erythropoiesis may prove beneficial by increasing blood viscosity and shear stress, leading to vasodilatation, in addition to the increase in oxygen-carrying capacity. Calculations show that oxygen extraction in the tissue of the window chamber model was significantly lowered in adapted animals breathing 5% oxygen, but was unchanged from the control when breathing 21% oxygen, even though blood hemoglobin content was increased from 14.5 +/- 0.07 g/dl at control to 21.04 +/- 1.24 g/dl in the adapted animals (P < 0.05). PMID- 14561681 TI - Age-related sensitivity to nicotine for inducible atrial tachycardia and atrial fibrillation. AB - The influence of nicotine in modulating vulnerability to atrial tachycardia and fibrillation (AT/AF) remains ill defined. The isolated hearts of six young (2-3 mo) and six old (22-24 mo) male Fischer 344 rats were Langendorff perfused at 5 ml/min with oxygenated Tyrode solution at 37 degrees C, and the whole heart was also super-fused with warmed oxygenated Tyrode solution at 15 ml/min. Nicotine prolonged the interatrial conduction time and effective refractory period that were significantly (P < 0.05) higher in the old than in the young rats in a concentration-dependent manner. Nicotine had a biphasic effect on burst atrial pacing-induced AT in both groups, increasing it at 10-30 ng/ml while decreasing it at 50-100 ng/ml (P < 0.01). Nicotine at 10-100 ng/ml increased burst atrial pacing-induced AF in the young rats but suppressed it in the old rats (P < 0.01). Optical mapping showed the presence of multiple independent wavefronts during AF and a single periodic large wavefront during AT in both groups. Nicotine, at concentrations found in the blood of smokers (30-85 ng/ml), exerts biphasic effects on inducible AT/AF in young rats and suppresses it in the old rats by causing high degrees of interatrial conduction block. PMID- 14561682 TI - Hydrolysis of Ca2+-sensitive fluorescent probes by perfused rat heart. AB - Rat hearts were loaded with the fluorescent calcium indicators fura 2, indo 1, rhod 2, or fluo 3 to determine cytosolic calcium levels in the perfused rat heart. With fura 2, however, basal tissue fluorescence increased above anticipated levels, suggesting accumulation of intermediates of fura 2-AM deesterification. To examine this process, we separated the intermediates of the deesterification process using HPLC after incubation of fura 2-AM with tissue homogenates and after loading in the rat heart. Loading of hearts with fura 2-AM resulted in tissue levels of fura 2 free acid that were only 5% of the total heart dye content of all fura 2 species. The parent fura 2-AM form accumulated without accumulation of intermediate products. Similar results were obtained with indo 1-AM. Fluo 3 loaded very poorly in perfused hearts. Unlike other indictors, rhod 2 rapidly loaded in perfused hearts and was completely converted to the free acid form. To determine the subcellular localization of the free acid form of these indictors, mitochondria from indicator-loaded hearts were assayed for the free acid form. Approximately 75% of the total amount of rhod 2 in hearts could be recovered in isolated mitochondria. Subcellular localization of indo 1 and fura 2 was more evenly distributed between mitochondria and nonmitochondrial compartments. We conclude that measurement of calcium in the perfused rat heart using surface fluorescence with either indo 1 or fura 2 is complicated by an inconsistent accumulation of the parent ester and that the resulting signal cannot be easily calibrated using "in situ" methods using the free acid form. Rhod 2 does not display this shortcoming, but like other indicators, it also loads into the mitochondrial matrix. PMID- 14561683 TI - Repeated inhalation of adrenomedullin ameliorates pulmonary hypertension and survival in monocrotaline rats. AB - Adrenomedullin (AM) is a potent vasodilator peptide. We investigated whether inhalation of aerosolized AM ameliorates monocrotaline (MCT)-induced pulmonary hypertension in rats. Male Wistar rats given MCT (MCT rats) were assigned to receive repeated inhalation of AM (n = 8) or 0.9% saline (n = 8). AM (5 mug/kg) or saline was inhaled as an aerosol using an ultrasonic nebulizer for 30 min four times a day. After 3 wk of inhalation therapy, mean pulmonary arterial pressure and total pulmonary resistance were markedly lower in rats treated with AM than in those given saline [mean pulmonary arterial pressure: 22 +/- 2 vs. 35 +/- 1 mmHg (-37%); total pulmonary resistance: 0.048 +/- 0.004 vs. 0.104 +/- 0.006 mmHg.ml(-1).min(-1).kg(-1) (-54%), both P < 0.01]. Neither systemic arterial pressure nor heart rate was altered. Inhalation of AM significantly attenuated the increase in medial wall thickness of peripheral pulmonary arteries in MCT rats. Kaplan-Meier survival curves demonstrated that MCT rats treated with aerosolized AM had a significantly higher survival rate than those given saline (70% vs. 10% 6-wk survival, log-rank test, P < 0.01). In conclusion, repeated inhalation of AM inhibited MCT-induced pulmonary hypertension without systemic hypotension and thereby improved survival in MCT rats. PMID- 14561684 TI - Antiinflammatory effects of omega-3 fatty acids vary at different stages of inflammation. PMID- 14561685 TI - Kinematic analysis of manual tracking in monkeys: characterization of movement intermittencies during a circular tracking task. AB - Segmentation of the velocity profiles into the submovements has been observed in reaching and tracking limb movements and even in isometric tasks. Submovements have been implicated in both feed-forward and feedback control. In this study, submovements were analyzed during manual tracking in the nonhuman primate with the focus on the amplitude-duration scaling of submovements and the error signals involved in their control. The task consisted of the interception and visually guided pursuit of a target moving in a circle. The submovements were quantified based on their duration and amplitude in the speed profile. Control experiments using passive movements demonstrated that these intermittencies were not instrumentation artifacts. Submovements were prominent in both the interception and tracking phases and their amplitude scaled linearly with duration. The scaling factors increased with tracking speed at the same rate for both interception and pursuit. A cross-correlation analysis between a variety of error signals and the speed profile revealed that direction and speed errors were temporally coupled to the submovements. The cross-correlation profiles suggest that submovements are initiated when speed error reaches a certain limit and when direction error is minimized. The scaling results show that in monkeys submovements characterize both the interception and pursuit portions of the task and that these submovements have similar scaling properties consistent with 1) the concept of stereotypy and 2) adding constant acceleration/force at a specific tracking speed. The correlation results show involvement of speed and direction error signals in controlling the submovements. PMID- 14561686 TI - Signals that modulate gain control for smooth pursuit eye movements in monkeys. AB - The generation of primate smooth pursuit eye movements involves two processes. One process transforms the direction and speed of target motion into a motor command and the other regulates the strength, or "gain," of the visual-motor transformation. We have conducted a behavioral analysis to identify the signals that modulate the internal gain of pursuit. To test whether the modulatory signals are related to eye velocity in the orbit or in the world (gaze velocity), we used brief perturbations of target motion to probe the gain of pursuit during tracking conditions that used head rotation to dissociate eye and gaze velocity. We found that the responses to perturbations varied primarily as a function of gaze velocity. To further understand the gaze velocity signals that control internal pursuit gain, we used adaptive modification of the gain of the vestibulo ocular reflex (VOR) to dissociate physical gaze velocity from the component of gaze velocity that is driven by visual inputs. After VOR adaptation, perturbation responses were altered; the smallest perturbation responses now occurred during tracking conditions that required nonzero physical gaze velocity. However, perturbation responses during tracking conditions that mimicked the modified VOR were still enhanced relative to those obtained during fixation. We conclude that the signals that modulate the internal gain of pursuit are modified by VOR adaptation so that they are rendered intermediate between physical and visually driven gaze velocity. Similar changes in the gaze velocity signal have been reported in the cerebellar floccular complex following adaptive modification of the VOR and could be present in other brain areas that carry putative gaze velocity signals. PMID- 14561687 TI - The role of execution noise in movement variability. AB - The origin of variability in goal-directed movements is not well understood. Variability can originate from several neural processes such as target localization, movement planning, and movement execution. Here we examine variability resulting from noise in movement execution. In several experiments, subjects moved their unseen hand to visual targets, under conditions which were designed to minimize the variability expected from localization and planning processes. We tested short movements in 32 directions in a center-out reaching task. The variability in the movement endpoints and in the initial movement direction varied systematically with the movement direction, with some directions having up to twice the variability of others. In a second experiment we tested four movements in the same direction but with different extents. Here, the longer movements were systematically curved, and the endpoint ellipses were not aligned with the straight line between starting and end position, but they were roughly aligned with the last part of the trajectory. We show that the variability observed in these experiments cannot be explained by planning noise but is well explained by noise in movement execution. A combination of both signal-dependent and signal-independent noise in the amplitude of the motor commands and temporal noise in their duration can explain the observed variability. Our results suggest that, in general, execution noise accounts for at least a large proportion of movement variability. PMID- 14561688 TI - Endogenous zinc inhibits GABA(A) receptors in a hippocampal pathway. AB - Depending on their subunit composition, GABA(A) receptors can be highly sensitive to Zn(2+). Although a pathological role for Zn(2+)-mediated inhibition of GABA(A) receptors has been postulated, no direct evidence exists that endogenous Zn(2+) can modulate GABAergic signaling in the brain. A possible explanation is that Zn(2+) is mainly localized to a subset of glutamatergic synapses. Hippocampal mossy fibers are unusual in that they are glutamatergic but have also been reported to contain GABA and Zn(2+). Here, we show, using combined Timm's method and post-embedding immunogold, that the same mossy fiber varicosities can contain both GABA and Zn(2+). Chelating Zn(2+) with either calcium-saturated EDTA or N,N,N',N'-tetrakis (2-pyridylmethyl)ethylenediamine had no effect on stratum radiatum-evoked inhibitory postsynaptic currents (IPSCs), but enhanced IPSCs evoked by stimuli designed to recruit dentate granule cells. We also show that IPSCs recorded in CA3 pyramidal neurons in acute hippocampal slices are depressed by exogenous Zn(2+). This depression was of similar amplitude whether the IPSCs were evoked by stimulation in s. radiatum (to recruit local interneurons) or in the s. granulosum of the dentate gyrus (to recruit mossy fibers). These results show for the first time that GABAergic IPSCs can be modulated by endogenous Zn(2+) and are consistent with GABA release at Zn(2+)-containing mossy fiber synapses. PMID- 14561689 TI - Involvement of the ryanodine receptor in morphologic modification of Hermissenda type B photoreceptors after in vitro conditioning. AB - We examined whether Ca(2+) induced Ca(2+) release through ryanodine receptors is involved in the conditioning of specific morphologic changes at the axon terminals of type B photoreceptors in the isolated circumesophageal ganglion of Hermissenda. Calcium chelation by bis(2-aminophenoxy) ethane-N,N,N',N' tetraacetic acid prevented the conformational change at the terminals after five paired presentations of light and vibration, which produce terminal branch contraction of B photoreceptors. Two ryanodine receptor blockers, dantrolene and micromolar concentrations of ryanodine, depressed the increase in excitability due to in vitro conditioning and the increase in intracellular Ca(2+) in response to membrane depolarization. Although the ability to increase intracellular Ca(2+) was depressed, synaptic transmission was preserved in the normal state from hair cells under dantrolene and ryanodine incubation. Ryanodine receptor blockers also prevented contraction at the B photoreceptor axon terminals. These results suggest that the ryanodine receptor has a crucial role in inducing the in vitro conditioning specific changes both physiologically and morphologically, including "focusing" at the B photoreceptor axon terminal. PMID- 14561690 TI - Differences in glycinergic mIPSCs in the auditory brain stem of normal and congenitally deaf neonatal mice. AB - We have investigated the fundamental properties of central auditory glycinergic synapses in early postnatal development in normal and congenitally deaf (dn/dn) mice. Glycinergic miniature inhibitory postsynaptic currents (mIPSCs) were recorded using patch-clamp methods in neurons from a brain slice preparation of the medial nucleus of the trapezoid body (MNTB), at 12-14 days postnatal age. Our results show a number of significant differences between normal and deaf mice. The frequency of mIPSCs is greater (50%) in deaf versus normal mice. Mean mIPSC amplitude is smaller in deaf mice than in normal mice (mean mIPSC amplitude: deaf, 64 pA; normal, 106 pA). Peak-scaled fluctuation analysis of mIPSCs showed that mean single channel conductance is greater in the deaf mice (deaf, 64 pS; normal, 45 pS). The mean decay time course of mIPSCs is slower in MNTB neurons from deaf mice (mean half-width: deaf, 2.9 ms; normal, 2.3 ms). Light- and electron-microscopic immunolabeling results showed that MNTB neurons from deaf mice have more (30%) inhibitory synaptic sites (postsynaptic gephyrin clusters) than MNTB neurons in normal mice. Our results demonstrate substantial differences in glycinergic transmission in normal and congenitally deaf mice, supporting a role for activity during development in regulating both synaptic structure (connectivity) and the fundamental (quantal) properties of mIPSCs at central glycinergic synapses. PMID- 14561691 TI - Properties of saccadic responses in monkey when multiple competing visual stimuli are present. AB - Important insights into the neural organization of the saccadic system have been gained when the usually stereotyped movement trajectories of saccades have been altered by experimental manipulation. In the present study we produced trajectory variability in monkeys by using a visual search task in which both the location and color of an odd-colored target were changed randomly trial by trial, and the number of distractors was varied on each trial. We wished to determine whether increasing the number of distractors also increased the movement trajectory variation, i.e., the amount of initial directional deviation, endpoint deviation (averaging), and curvature of saccades. Overall, saccade latencies and the proportion of saccades directed to distractors decreased as the number of homogenous distractors increased. We also found that saccades have much more dispersion in their initial direction when distractors are present in comparison to the case when only a single target without distractors appears. However, initial dispersion decreases systematically as the number of distractors increases. The percentage of averaging saccades produced in the search task was not consistently dependent on the number of distractors. A significant fraction of averaging saccades still occurred for much wider spatial separations of stimuli than in previous studies using two visual stimuli with no specified target. The curvature of saccade trajectories increased dramatically when distractors were present, but the amount of curvature was not systematically affected by the number of distractors. Errors present in saccade trajectory in the search task were only poorly compensated. We conclude that these variable saccade trajectories result from incomplete or inaccurate specification of the target when competing stimuli are present and that a smaller number of more widely spread distractors facilitate saccade variability, perhaps due to the greater difficulty of target selection. PMID- 14561692 TI - Nasal trigeminal inputs release the A5 inhibition received by the respiratory rhythm generator of the mouse neonate. AB - Experiments were performed on neonatal mice to analyze why, in vitro, the respiratory rhythm generator (RRG) was silent and how it could be activated. We demonstrated that in vitro the RRG in intact brain stems is silenced by a powerful inhibition arising from the pontine A5 neurons through medullary alpha(2) adrenoceptors and that in vivo nasal trigeminal inputs facilitate the RRG as nasal continuous positive airway pressure increases the breathing frequency, whereas nasal occlusion and nasal afferent anesthesia depress it. Because nasal trigeminal afferents project to the A5 nuclei, we applied single trains of negative electric shocks to the trigeminal nerve in inactive ponto medullary preparations. They induced rhythmic phrenic bursts during the stimulation and for 2-3 min afterward, whereas repetitive trains produced on going rhythmic activity up to the end of the experiments. Electrolytic lesion or pharmacological inactivation of the ipsilateral A5 neurons altered both the phrenic burst frequency and occurrence after the stimulation. Extracellular unitary recordings and trans-neuronal tracing experiments with the rabies virus show that the medullary lateral reticular area contains respiratory-modulated neurons, not necessary for respiratory rhythmogenesis, but that may provide an excitatory pathway from the trigeminal inputs to the RRG as their electrolytic lesion suppresses any phrenic activity induced by the trigeminal nerve stimulation. The results lead to the hypothesis that the trigeminal afferents in the mouse neonate involve at least two pathways to activate the RRG, one that may act through the medullary lateral reticular area and one that releases the A5 inhibition received by the RRG. PMID- 14561693 TI - The yin and yang of rheumatoid arthritis. PMID- 14561695 TI - Neurogenesis and brain injury: managing a renewable resource for repair. AB - The brain shows limited ability to repair itself, but neurogenesis in certain areas of the adult brain suggests that neural stem cells may be used for structural brain repair. It will be necessary to understand how neurogenesis in the adult brain is regulated to develop strategies that harness neural stem cells for therapeutic use. PMID- 14561696 TI - What brings pericytes to tumor vessels? AB - Paracrine signaling via platelet-derived growth factor B (PDGFB), expressed by endothelial cells, and its receptor PDGFR-beta, expressed by pericytes, plays a central role in blood vessel maturation. A new study (see the related article beginning on page 1142) reveals that it is not just the presence of PDGFB, but how it is presented to pericytes, that determines the quality of the endothelium pericyte interaction. PMID- 14561697 TI - Receptor "cross talk" in innate immunity. AB - Toll-like receptors (TLRs) recognize microbial molecular signatures and can initiate innate immune responses against invading pathogens. A new study (see the related article beginning on page 1234) reports how TLR2 expression by endothelia is locally upregulated by the action of activated polymorphonuclear neutrophils via an unprecedented mechanism involving cell-cell interaction and NAD(P)H oxidase. The report reveals yet another way in which the primordial innate immune system is remarkably complex. PMID- 14561698 TI - A novel role for uroguanylin in the regulation of sodium balance. AB - Uroguanylin is a peptide hormone that regulates sodium excretion by the kidney when excess NaCl is consumed. A new study demonstrates that mice deficient in uroguanylin have blunted urinary sodium excretion responses to oral sodium loads in addition to elevated blood pressure (see related article beginning on page 1244). A physiological role for uroguanylin is discussed, linking the intestine and kidney in an endocrine axis for the maintenance of sodium balance. PMID- 14561699 TI - Endothelial and nonendothelial sources of PDGF-B regulate pericyte recruitment and influence vascular pattern formation in tumors. AB - Tumor-infiltrating blood vessels deviate morphologically and biochemically from normal vessels, raising the prospect of selective pharmacological targeting. Current antiangiogenic approaches focus mainly on endothelial cells, but recent data imply that targeting pericytes may provide additional benefits. Further development of these concepts will require deeper insight into mechanisms of pericyte recruitment and function in tumors. Here, we applied genetic tools to decipher the function of PDGF-B and PDGF-Rbeta in pericyte recruitment in a mouse fibrosarcoma model. In tumors transplanted into PDGF-B retention motif-deficient (pdgf-b(ret/ret)) mice, pericytes were fewer and were partially detached from the vessel wall, coinciding with increased tumor vessel diameter and hemorrhaging. Transgenic PDGF-B expression in tumor cells was able to increase the pericyte density in both WT and pdgf-b(ret/ret) mice but failed to correct the pericyte detachment in pdgf-b(ret/ret) mice. Coinjection of exogenous pericytes and tumor cells showed that pericytes require PDGF-Rbeta for recruitment to tumor vessels, whereas endothelial PDGF-B retention is indispensable for proper integration of pericytes in the vessel wall. Our data support the notion that pericytes serve an important function in tumor vessels and highlight PDGF-B and PDGF-Rbeta as promising molecular targets for therapeutic intervention. PMID- 14561700 TI - A population of c-Kit(low)(CD45/TER119)- hepatic cell progenitors of 11-day postcoitus mouse embryo liver reconstitutes cell-depleted liver organoids. AB - Embryo liver morphogenesis takes place after gastrulation and starts with a ventral foregut evagination that reacts to factor signaling from both cardiac mesoderm and septum transversum mesenchyme. Current knowledge of the progenitor stem cell populations involved in this early embryo liver development is scarce. We describe here a population of 11-day postcoitus c-Kit(low)(CD45/TER119)- liver progenitors that selectively expressed hepatospecific genes and proteins in vivo, was self-maintained in vitro by long-term proliferation, and simultaneously differentiated into functional hepatocytes and bile duct cells. Purified c Kit(low)(CD45/TER119)- liver cells cocultured with cell-depleted fetal liver fragments engrafted and repopulated the hepatic cell compartments of the latter organoids, suggesting that they may include the embryonic stem cells responsible for liver development. PMID- 14561702 TI - Inhibition of NF-kappaB activation in macrophages increases atherosclerosis in LDL receptor-deficient mice. AB - Atherosclerosis is now generally accepted as a chronic inflammatory condition. The transcription factor NF-kappaB is a key regulator of inflammation, immune responses, cell survival, and cell proliferation. To investigate the role of NF kappaB activation in macrophages during atherogenesis, we used LDL receptor deficient mice with a macrophage-restricted deletion of IkappaB kinase 2 (IKK2), which is essential for NF-kappaB activation by proinflammatory signals. These mice showed increased atherosclerosis as quantified by lesion area measurements. In addition, the lesions were more advanced and showed more necrosis and increased cell number in early lesions. Southern blotting revealed that deletion of IKK2 was approximately 65% in macrophages, coinciding with a reduction of 50% in NF-kappaB activation, as compared with controls. In both groups, the expression of differentiation markers, uptake of bacteria, and endocytosis of modified LDL was similar. Upon stimulation with LPS, production of TNF was reduced by approximately 50% in IKK2-deleted macrophages. Interestingly, we also found a major reduction in the anti-inflammatory cytokine IL-10. Our data show that inhibition of the NF-kappaB pathway in macrophages leads to more severe atherosclerosis in mice, possibly by affecting the pro- and anti-inflammatory balance that controls the development of atherosclerosis. PMID- 14561701 TI - Antibodies to a cell surface histone-like protein protect against Histoplasma capsulatum. AB - A protective role for antibodies has not previously been described for host defense against the pathogenic fungus Histoplasma capsulatum (Hc). Mouse mAb's were generated from mice immunized with Hc yeast that binds the cell surface of Hc. Administration of mAb's before Hc infection reduced fungal burden, decreased pulmonary inflammation, and prolonged survival in a murine infection model. Protection mediated by mAb's was associated with enhanced levels of IL-4, IL-6, and IFN-gamma in the lungs of infected mice. The mAb's increased phagocytosis of yeast by J774.16 cells through a CR3-dependent process. Ingestion of mAb opsonized Hc by J774.16 macrophage-like cells was associated with yeast cell growth inhibition and killing. The mAb's bound to a 17-kDa antigen expressed on the surface of Hc. The antigen was identified as a histone H2B-like protein. This study establishes that mAb's to a cell surface protein of Hc alter the intracellular fate of the fungus and mediate protection in a murine model of lethal histoplasmosis, and it suggests a new candidate antigen for vaccine development. PMID- 14561703 TI - IL-23 produced by CNS-resident cells controls T cell encephalitogenicity during the effector phase of experimental autoimmune encephalomyelitis. AB - CNS-resident cells, in particular microglia and macrophages, are a source of inflammatory cytokines during inflammation within the CNS. Expression of IL-23, a recently discovered cytokine, has been shown to be critical for the development of experimental autoimmune encephalomyelitis (EAE) in mice. Expression of the p40 subunit of IL-12 and IL-23 by microglia has been shown in situ and in vitro, but direct evidence for a functional significance of p40 expression by CNS cells during an immune response in vivo is still lacking. Here we report that p40 plays a critical role in maintaining encephalitogenicity during the disease course. By using irradiation bone marrow chimeras, we have generated mice in which p40 is deleted from the CNS parenchyma but not the systemic immune compartment. Our studies show that p40 expressed by CNS-endogenous cells is critical for the development of myelin oligodendrocyte glycoprotein-induced EAE. In spite of the reduced clinical disease, the absence of p40 from the CNS has little impact on the degree of inflammation. Expression profiles of the CNS lesions show an increase in Th2 cytokines when compared with mice that develop EAE in the presence of CNS IL-12 and/or IL-23. Taken together, our data demonstrate that p40 expression by CNS-resident cells forms the basis for the Th1 bias of the CNS. PMID- 14561704 TI - A homozygous mutation in HESX1 is associated with evolving hypopituitarism due to impaired repressor-corepressor interaction. AB - The paired-like homeobox gene expressed in embryonic stem cells Hesx1/HESX1 encodes a developmental repressor and is expressed in early development in a region fated to form the forebrain, with subsequent localization to Rathke's pouch, the primordium of the anterior pituitary gland. Mutations within the gene have been associated with septo-optic dysplasia, a constellation of phenotypes including eye, forebrain, and pituitary abnormalities, or milder degrees of hypopituitarism. We identified a novel homozygous nonconservative missense mutation (I26T) in the critical Engrailed homology repressor domain (eh1) of HESX1, the first, to our knowledge, to be described in humans, in a girl with evolving combined pituitary hormone deficiency born to consanguineous parents. Neuroimaging revealed a thin pituitary stalk with anterior pituitary hypoplasia and an ectopic posterior pituitary, but no midline or optic nerve abnormalities. This I26T mutation did not affect the DNA-binding ability of HESX1 but led to an impaired ability to recruit the mammalian Groucho homolog/Transducin-like enhancer of split-1 (Gro/TLE1), a crucial corepressor for HESX1, thereby leading to partial loss of repression. Thus, the novel pituitary phenotype highlighted here appears to be a specific consequence of the inability of HESX1 to recruit Groucho-related corepressors, suggesting that other molecular mechanisms govern HESX1 function in the forebrain. PMID- 14561705 TI - FGF-2 regulates neurogenesis and degeneration in the dentate gyrus after traumatic brain injury in mice. AB - We studied the role of FGF-2 on regulation of neurogenesis and cell loss in the granule cell layer (GCL) of the hippocampal dentate gyrus after experimental traumatic brain injury (TBI). In both FGF-2(-/-) and FGF-2(+/+) mice subjected to controlled cortical impact, the number of dividing cells labeled with BrdU, injected on posttrauma days 6 through 8, increased at 9 days after TBI, and the number of BrdU-positive cells colabeled with neuron-specific nuclear antigen significantly increased at 35 days. However, in injured FGF-2-/- mice, BrdU positive cells and BrdU-positive neurons (days 9, 35) were fewer compared with FGF-2(+/+) mice. There was also a decrease in the volume of the GCL and the number of GCL neurons after TBI in both FGF-2(-/-) and FGF-2(+/+) mice, but the decrease in both was greater in FGF-2-/- mice at 35 days. Overexpression of FGF-2 by intracerebral injection of herpes simplex virus-1 amplicon vectors encoding this factor increased numbers of dividing cells (day 9) and BrdU-positive neurons (day 35) significantly in C57BL/6 mice. Furthermore, the decrease in GCL volume was also attenuated. These results suggest that FGF-2 upregulates neurogenesis and protects neurons against degeneration in the adult hippocampus after TBI, and that FGF-2 supplementation via gene transfer can reduce GCL degeneration after TBI. PMID- 14561706 TI - Activation of natural killer T cells in NZB/W mice induces Th1-type immune responses exacerbating lupus. AB - In vivo treatment of mice with the natural killer T (NKT) cell ligand, alpha galactosylceramide (alphaGalCer), ameliorates autoimmune diabetes and experimental autoimmune encephalomyelitis (EAE) by shifting pathogenic Th1-type immune responses to nonpathogenic Th2-type responses. In the current study, in vivo activation of NKT cells in adult NZB/W mice by multiple injections of alphaGalCer induced an abnormal Th1-type immune response as compared with the Th2 type response observed in nonautoimmune C57BL/6 mice. This resulted in decreased serum levels of IgE, increased levels of IgG2a and IgG2a anti-double-stranded DNA (anti-dsDNA) Ab's, and exacerbated lupus. Conversely, treatment of NZB/W mice with blocking anti-CD1d mAb augmented Th2-type responses, increased serum levels of IgE, decreased levels of IgG2a and IgG2a anti-dsDNA Ab's, and ameliorated lupus. While total CD4+ T cells markedly augmented in vitro IgM anti-dsDNA Ab secretion by splenic B cells, the non-CD1d-reactive (CD1d-alphaGalCer tetramer negative) CD4+ T cells (accounting for 95% of all CD4+ T cells) failed to augment Ab secretion. The CD1d-reactive tetramer-positive CD4+ T cells augmented anti dsDNA Ab secretion about tenfold. In conclusion, activation of NKT cells augments Th1-type immune responses and autoantibody secretion that contribute to lupus development in adult NZB/W mice, and anti-CD1d mAb might be useful for treating lupus. PMID- 14561707 TI - Loss of Tsc1/Tsc2 activates mTOR and disrupts PI3K-Akt signaling through downregulation of PDGFR. AB - Tuberous sclerosis (TSC) is a familial tumor syndrome due to mutations in TSC1 or TSC2, in which progression to malignancy is rare. Primary Tsc2(-/-) murine embryo fibroblast cultures display early senescence with overexpression of p21CIP1/WAF1 that is rescued by loss of TP53. Tsc2(-/-)TP53(-/-) cells, as well as tumors from Tsc2(+/-) mice, display an mTOR-activation signature with constitutive activation of S6K, which is reverted by treatment with rapamycin. Rapamycin also reverts a growth advantage of Tsc2(-/-)TP53(-/-) cells. Tsc1/Tsc2 does not bind directly to mTOR, however, nor does it directly influence mTOR kinase activity or cellular phosphatase activity. There is a marked reduction in Akt activation in Tsc2(-/ )TP53(-/-) and Tsc1(-/-) cells in response to serum and PDGF, along with a reduction in cell ruffling. PDGFRalpha and PDGFRbeta expression is markedly reduced in both the cell lines and Tsc mouse renal cystadenomas, and ectopic expression of PDGFRbeta in Tsc2-null cells restores Akt phosphorylation in response to serum, PDGF, EGF, and insulin. This activation of mTOR along with downregulation of PDGFR PI3K-Akt signaling in cells lacking Tsc1 or Tsc2 may explain why these genes are rarely involved in human cancer. This is in contrast to PTEN, which is a negative upstream regulator of this pathway. PMID- 14561708 TI - TLR4 signaling induces TLR2 expression in endothelial cells via neutrophil NADPH oxidase. AB - Interactions of polymorphonuclear neutrophils (PMNs) with endothelial cells may contribute to the activation of endothelial cell responses involved in innate immunity. We explored a novel function of PMN NADPH oxidase in the mechanism of Toll-like receptor-2 (TLR2) upregulation induced by LPS-TLR4 signaling in endothelial cells. We showed that LPS induced TLR2 up-regulation through TLR4- and MyD88-dependent signaling. In neutropenic mice, the LPS-induced NF-kB activation and TLR2 expression were significantly reduced, and both responses were restored upon repletion by PMN obtained from WT mice but not by PMNs from NADPH oxidase gp91pho(-/-) mice. These findings were recapitulated in mouse lung vascular endothelial cells cocultured with PMNs, indicating that the augmented NF kB activation and the resultant TLR2 upregulation in endothelial cells were secondary to oxidant signaling generated by PMN NADPH oxidase. The functional relevance of NADPH oxidase in mediating TLR4-induced TLR2 expression in endothelial cells was evident by markedly elevated and stable ICAM-1 expression as well as augmented PMN migration in response to sequential challenge with LPS and peptidoglycan. Thus, PMN NADPH oxidase-derived oxidant signaling is an important determinant of the cross talk between TLR4 and TLR2 and the control of endothelial cell activation. PMID- 14561709 TI - Uroguanylin knockout mice have increased blood pressure and impaired natriuretic response to enteral NaCl load. AB - Guanylin and uroguanylin, peptides synthesized in the intestine and kidney, have been postulated to have both paracrine and endocrine functions, forming a potential enteric-renal link to coordinate salt ingestion with natriuresis. To explore the in vivo role of uroguanylin in the regulation of sodium excretion, we created gene-targeted mice in which uroguanylin gene expression had been ablated. Northern and Western analysis confirmed the absence of uroguanylin message and protein in knockout mice, and cGMP levels were decreased in the mucosa of the small intestine. Ussing chamber analysis of jejunum revealed that Na+/H+ exchanger-mediated Na+ absorption and tissue conductance was not altered in the knockout animals, but short-circuit current, an index of electrogenic anion secretion, was reduced. Renal clearance measurements showed that uroguanylin deficiency results in impaired ability to excrete an enteral load of NaCl, primarily due to an inappropriate increase in renal Na+ reabsorption. Finally, telemetric recordings of blood pressure demonstrated increased mean arterial pressure in uroguanylin knockout animals that was independent of the level of dietary salt intake. Together, these findings establish a role for uroguanylin in an enteric-renal communication axis as well as a fundamental principle of this axis in the maintenance of salt homeostasis in vivo. PMID- 14561712 TI - Retraction. PMID- 14561710 TI - Autosomal dominant pseudohypoparathyroidism type Ib is associated with a heterozygous microdeletion that likely disrupts a putative imprinting control element of GNAS. AB - Patients with pseudohypoparathyroidism type Ib (PHP-Ib) have hypocalcemia and hyperphosphatemia due to renal parathyroid hormone (PTH) resistance, but lack physical features of Albright hereditary osteodystrophy. PHP-Ib is thus distinct from PHP-Ia, which is caused by mutations in the GNAS exons encoding the G protein alpha subunit. However, an imprinted autosomal dominant form of PHP-Ib (AD-PHP-Ib) has been mapped to a region of chromosome 20q13.3 containing GNAS. Furthermore, loss of methylation at a differentially methylated region (DMR) of this locus, exon A/B, has been observed thus far in all investigated sporadic PHP Ib cases and the affected members of multiple AD-PHP-Ib kindreds. We now report that affected members and obligate gene carriers of 12 unrelated AD-PHP-Ib kindreds and four apparently sporadic PHP-Ib patients, but not healthy controls, have a heterozygous approximately 3-kb microdeletion located approximately 220 kb centromeric of GNAS exon A/B. The deleted region, which is flanked by two direct repeats, includes three exons of STX16, the gene encoding syntaxin-16, for which no evidence of imprinting could be found. Affected individuals carrying the microdeletion show loss of exon A/B methylation but no epigenetic abnormalities at other GNAS DMRs. We therefore postulate that this microdeletion disrupts a putative cis-acting element required for methylation at exon A/B, and that this genetic defect underlies the renal PTH resistance in AD-PHP-Ib. PMID- 14561713 TI - Signal transduction underlying cell morphogenesis: editorial overview. PMID- 14561714 TI - Regulation of actin dynamics by WASP family proteins. AB - Rapid reorganization of the actin cytoskeleton underlies morphological changes and motility of cells. WASP family proteins have received a great deal of attention as the signal-regulated molecular switches that initiate actin polymerization. The first member, WASP, was identified as the product of a gene of which dysfunction causes the human hereditary disease Wiskott-Aldrich syndrome. There are now five members in this protein family, namely WASP, N-WASP, WAVE/Scar1, 2, and 3. WASP and N-WASP have functional and physical associations with Cdc42, a Rho family small GTPase involved in filopodium formation. In contrast, there is evidence that links the WAVE/Scar proteins with another Rho family protein, Rac, which is a regulator of membrane ruffling. All WASP family members have a VCA domain at the C-terminus through which Arp2/3 complex is activated to nucleate actin polymerization. Analyses of model organisms have just begun to reveal unexpected functions of WASP family proteins in multicellular organisms. PMID- 14561715 TI - Dynamic control of neuronal morphogenesis by rho signaling. AB - Polarization of the neuronal cell body and initiation of the first neuritic process represent the starting point of a series of dynamic metamorphic events by which the newly acquired identity of a group of neurons can be translated into a morphologically complex web of three-dimensional neuronal circuit. Despite the critical importance of these events, little is known about the molecular signaling mechanisms that either regulate the temporal sequence of these steps or ensure the accuracy and the spatial consistency of the resulting circuits. In this review, based on recent findings from our group and others, we present a working model on how the initial events in neuronal morphogenesis in the CNS may be controlled by multiple Rho pathways. PMID- 14561716 TI - "Search-and-capture" of microtubules through plus-end-binding proteins (+TIPs). AB - The generation of a polarized microtubule organization is critically important for proper cellular functions, such as cell division, differentiation and migration. Microtubules themselves are highly dynamic structures, and this dynamic property is temporally and spatially regulated within cells, especially at their plus ends. To explain how microtubules set up and make contacts with cellular structures, a "search-and-capture" mechanism has been proposed, in which the microtubule plus ends dynamically search for and capture specific sites, such as mitotic kinetochores and cell cortex. To date, several classes of proteins have been shown to be associated with microtubule plus ends in a wide range of organisms from fungi to humans and to play critical roles in the "search-and capture" mechanism. In this review, we overview our current understanding of the "plus-end-binding proteins" (+TIPs), including APC (adenomatous polyposis coli) tumor suppressor protein, cytoplasmic linker proteins (CLIPs), CLIP-associating proteins (CLASPs), cytoplasmic dynein/dynactin, and EB1, an APC-interacting protein. PMID- 14561717 TI - Structural basis of the Rho GTPase signaling. AB - Small GTPases of the Rho family serve as conformational switches in a wide variety of signal transduction pathways that regulate diverse cellular functions. The GTP-bound forms of Rho GTPases are capable of interacting with downstream effectors that control cytoskeletal rearrangements. Regulators that stimulate nucleotide exchange, the hydrolytic cycle and distribution between the membrane and cytosol control the switch. Detailed pictures of Rho GTPase switching, effector recognition and regulation by regulators have emerged from recent structural investigations. These include the most extensively studied Rho GTPases, RhoA, Rac1, 2 and Cdc42, and their complexes with effectors and regulators. These studies have revealed the general diversity of effector and regulator structures, and in particular the structural features concerning the specific interactions involved in Rho effector recognition and regulator interactions with Rho GTPase. These findings provide a critical insight into the nature of Rho GTPase activity and consequently allow for a detailed manipulation of signaling pathways mediated by these proteins. PMID- 14561718 TI - Two mitofusin proteins, mammalian homologues of FZO, with distinct functions are both required for mitochondrial fusion. AB - Mitochondria are dynamic organelles that undergo frequent fission and fusion or branching. Although these morphologic changes are considered crucial for cellular functions, the underlying mechanisms remain elusive, especially in mammalian cells. We characterized two rat mitochondrial outer membrane proteins, Mfn1 and Mfn2, with distinct tissue expressions, that are homologous to Drosophila Fzo, a GTPase involved in mitochondrial fusion. Expression of the GTPase-domain mutant of Mfn2 (Mfn2(K109T)) in HeLa cells induced mitochondrial fragmentation in which Mfn2(K109T) localized at the restricted domains. Immuno-electronmicroscopy revealed that Mfn2(K109T) was concentrated at the contact domains between adjacent mitochondria, suggesting that fusion of the outer membrane was arrested at some intermediate step. Mfn1 expression induced highly connected tubular network structures depending on the functional GTPase domain. The Mfn1-induced tubular networks were suppressed by co-expression with Mfn2. In vivo depletion of either isoform by RNA interference revealed that both are required to maintain normal mitochondrial morphology. The fusion of differentially-labeled mitochondria in HeLa cells subjected to depletion of either Mfn isoform and subsequent cell fusion by hemagglutinating virus of Japan revealed that both proteins have distinct functions in mitochondrial fusion. We conclude that the two Mfn isoforms cooperate in mitochondrial fusion in mammalian cells. PMID- 14561719 TI - Variation in the divalent cation requirements of influenza a virus N2 neuraminidases. AB - Influenza virus N2 neuraminidases were chromatographically purified from several vaccine candidate strains from 1957 to 1994. Enzymatic kinetic parameters and immunogenicity were tested for each strain. For each NA tested, with ionic strength held constant, Ca(2+) or Mg(2+) increased the initial rate of enzymatic activity. Earlier N2-NA strains had the highest initial velocity, V(max)/K(m) and V(max). There were significant differences among the influenza virus strains in enzymatic activity before and after addition of Ca(2+) or Mg(2+): V(max)/K(m) varied from 0.54 M(-1) s(-1) to 0.88 M(-1) s(-1) and V(max) varied from 2.45 s( 1) to 4.3 s(-1) before the addition of a divalent cation; and increased approximately 2-fold each of these kinetic parameters for each strain after the addition of exogenous Ca(2+) or Mg(2+). Exhaustive dialysis with EDTA reduced the initial velocity of each strain with significant differences found among strains, with a range of 0.1% to 8% of original activity. Activity was partially restored by the addition of exogenous Ca(2+) or Mg(2+), varying from 8% to 60% of pre dialysis levels, but original rates were not achieved. This reduction in enzymatic activity for the tested strains (i.e., A/Japan/57 and A/Johannesburg/94) was accompanied by a parallel decrease in NA-immunogenicity, with antibody response decreasing by as much as 76% as measured by NI titer, and ELISA titer decreasing by as much as 68%. The addition of Ca(2+) or Mg(2+) to the post-dialysis sample restored immunogenicity to as much as 80% of pre-dialysis NI titers and as much as 78% of pre-dialysis ELISA titers. Dialysis had the least effect on early strains as measured by enzymatic kinetic parameters and immunogenicity studies. Zn(2+) had a slight inhibitory effect on the activity of all tested strains. Review of the nucleic acid sequence of each of these strains could not predict their enzymatic activity, immunogenicity or response to dialysis. If immunity against neuraminidase is desirable in vaccination against influenza, selection of vaccine candidate strains must include not only analysis of antigenic changes and sequence analysis but also enzymatic studies and determination of the requirement of divalent cations to maintain immunogenicity and activity during production. PMID- 14561720 TI - Enzyme-linked sensitive fluorometric imaging of glutamate release from cerebral neurons of chick embryos. AB - This paper describes a method for imaging the endogenous release of glutamate from cerebral neurons. This method is based on the reactions of glutamate oxidase and peroxidase, and on the detection of hydrogen peroxide by a fluorescent substrate of peroxidase. Glutamate has been sensitively measured in vitro in the range of 20 nM to 1 microM. We used two types of Ca(2+) channel inhibitors, MK 801 and omega-Conotoxin GVIA, which act to suppress Ca(2+) transport at postsynaptic and presynaptic neurons, respectively. MK-801 did not inhibit the increase in glutamate release after KCl stimulation, while there was no increase in glutamate release after KCl stimulation when omega-Conotoxin GVIA was used, probably due to the inhibition of voltage-activated Ca(2+) channels in the presynapse. Glutamate release and Ca(2+) flow in the synaptic regions were imaged using a laser confocal fluorescence microscope. KCl-evoked glutamate release was localized around cell bodies linked to axon terminals. This procedure allows imaging that can be sensitively detected by the fluorometric enzymatic assay of endogenous glutamate release in synapses. PMID- 14561721 TI - Vacuolar H+-ATPase inhibitor induces apoptosis via lysosomal dysfunction in the human gastric cancer cell line MKN-1. AB - We investigated the mechanism of apoptosis induced by bafilomycin A(1), an inhibitor of vacuolar H(+)-ATPase. Bafilomycin A(1) significantly inhibited the growth of MKN-1 human gastric cancer cells. Bafilomycin A(1) induced apoptosis as demonstrated by DNA ladder formation and the TUNEL method. We designed a flow cytometric assay to detect the alteration in lysosomal pH using a fluorescent probe, fluorescein isothiocyanate-conjugated dextran. This assay revealed that bafilomycin A(1) dramatically increased lysosomal pH. However, bafilomycin A(1) induced neither significant decrease in mitochondrial transmembrane potential nor the release of mitochondrial cytochrome c into the cytoplasm. Western blotting showed that cathepsin D, but not cathepsin L, was released into the cytoplasm. The activity of caspase-3 was significantly increased by bafilomycin A(1). However, cathepsin D did not directly cleave procaspase-3. These findings suggest that bafilomycin A(1)-induced apoptosis in MKN-1 cells is mediated by other proteases released after lysosomal dysfunction followed by activation of caspase 3 in a cytochrome c-independent manner. The present study showed that flow cytometric analysis of lysosomal pH can be useful to evaluate lysosomal protease mediated apoptosis. PMID- 14561722 TI - Rapid purification, characterization and substrate specificity of heparinase from a novel species of Sphingobacterium. AB - A type of heparinase (heparin lysase, no EC number) was isolated from the periplasmic space of a novel species of Sphingobacterium by three-step osmotic shock. It was further purified to apparent homogeneity by a combination of SP sepharose and Source 30S chromatographies with a final specific activity of 17.6 IU/mg protein and purification factor of 13-fold. MALDI-TOF mass spectrum of the purified heparinase gave a molecular mass of 75,674 Da of the native enzyme. Peptide mass spectrum showed poor homogeneity with the database in the peptide bank. Inhibition of the enzyme activity by N-acetylimidazole indicated that tyrosine residues were necessary for enzyme activity. K(m) and V(max) of the heparinase for de-o-sulfated-N-acetyl heparin were 42 micro M and 166 microM/min/mg protein, respectively. The heparinase showed similar activity on both heparin and heparan sulfate, except for the heparin from bovine lung. The heparinase exhibited only 8.3% of the activity when de-N-sulfated heparin was used as the substrate, but N-acetylation of the de-N-sulfated heparin restored the activity to 78.4%. Thus modification of N-site in heparin structure was favorable for heparinase activity. On the other hand, de-o-sulfation in heparin showed positive effects on the heparinase activity, since the enzyme activity for N-acetyl-de-o-sulfated heparin was increased by 150%. Based on the present findings, the sphingobacterial heparinase differed from flavobacterial and other reported heparinases in molecular mass, composition, charge properties, active site, substrate specificities and other important characteristics, suggesting that it a novel heparin lysase distinct from those from other sources. PMID- 14561723 TI - Response of genes associated with mitochondrial function to mild heat stress in yeast Saccharomyces cerevisiae. AB - The genome-wide expression pattern of budding yeast Saccharomyces cerevisiae in response to mild heat treatment in a non-fermentable carbon source was analyzed using DNA microarrays. Of 5,870 open reading frames (nuclear genome transcripts) examined, 104 genes were upregulated and 287 genes were downregulated upon shifting of the cells from 25 degrees C to 37 degrees C. Forty upregulated genes and 235 downregulated genes encoded localization-assigned proteins. Of 113 heat repressible genes (excluding 122 heat-repressible ribosomal genes), 36 were mitochondria-related genes, whereas only 2 of 40 heat-inducible genes were mitochondria-related. In particular, 9 genes involved in the mitochondrial respiratory chain and 7 genes involved in mitochondrial protein translocation were significantly repressed, suggesting that mitochondrial respiratory function and biogenesis were downregulated. Consistent with these findings, the growth of yeast cells in a non-fermentable carbon source was repressed at 37 degrees C and the mitochondria isolated from heat-stressed cells exhibited compromised preprotein-import activity compared with those from unstressed cells. In contrast, many genes involved in glycolysis and the metabolic pathway to produce glutamate via the tricarboxylic acid cycle, which is essential for biosynthetic reactions, were upregulated. Yeast cells might downregulate mitochondrial function to circumvent heat-induced oxidative stress, upregulate stress-related genes, and remodel genes for metabolic pathways in response to mild heat stress: an adaptive response at the expense of cell growth. PMID- 14561724 TI - Differential expression of two cathepsin Es during metamorphosis-associated remodeling of the larval to adult type epithelium in Xenopus stomach. AB - Cathepsin E (CE) was purified from the foregut of Xenopus laevis tadpoles as a mature dimeric form. The purified enzyme was a typical CE among aspartic proteinases with respect to pH dependence of proteolytic activity, susceptibility to pepstatin, and having N-linked high-mannose type oligosaccharide chains. We isolated two cDNAs for the CE (CE1 and CE2) from adult stomach. The amino acid sequence of the N-terminal region of the purified CE coincided with the corresponding sequence predicted from CE1. Northern blot analysis and in situ hybridization were performed. The CE1 mRNA was highly expressed in surface mucous cells and gland cells constituting the larval epithelium of the foregut of pro metamorphic tadpoles. As metamorphosis began and progressed, CE1 mRNA drastically decreased in amount, and subsequently both CE1 and CE2 mRNAs gradually increased. The increase in CE2 mRNA was detected shortly after the increase in CE1 mRNA. The decrease in CE1 expression correlated with degeneration of the larval type epithelium, while the increases in both CE1 and CE2 expression correlated with formation of the adult type epithelium. Thus, cathepsin E gene expression was differentially regulated during metamorphosis-associated remodeling of the larval to adult type epithelium in stomach. PMID- 14561725 TI - Characterization of functional domains of the hemolytic lectin CEL-III from the marine invertebrate Cucumaria echinata. AB - CEL-III is a Ca(2+)-dependent, galactose/N-acetylgalactosamine (GalNAc)-specific lectin isolated from the marine invertebrate Cucumaria echinata. This lectin exhibits strong hemolytic activity and cytotoxicity through pore formation in target cell membranes. The amino acid sequence of CEL-III revealed the N-terminal two-thirds to have homology to the B-chains of ricin and abrin, which are galactose-specific plant toxic lectins; the C-terminal one-third shows no homology to any known proteins. To examine the carbohydrate-binding ability of the N-terminal region of CEL-III, the protein comprising Pyr1-Phe283 was expressed in Escherichia coli cells. The expressed protein showed both the ability to bind to a GalNAc-immobilized column as well as hemagglutinating activity for rabbit erythrocytes, confirming that the N-terminal region has binding activity for specific carbohydrates. Since the C-terminal region could not be expressed in E. coli cells, a fragment containing this region was produced by limited proteolysis of the native protein by trypsin. The resulting C-terminal 15 kDa fragment of CEL-III exhibited a tendency to self-associate, forming an oligomer. When mixed with erythrocytes, the oligomer of the C-terminal fragment caused hemagglutination, probably due to hydrophobic interaction with cell membranes, while the monomeric fragment did not. Chymotryptic digestion of the preformed CEL-III oligomer induced upon lactose binding also yielded an oligomer of the C-terminal fragment comprising six molecules of the 16 kDa fragment. These results suggest that after binding to cell surface carbohydrate chains, CEL-III oligomerizes through C-terminal domains, leading to the formation of ion permeable pores by hydrophobic interaction with the cell membrane. PMID- 14561726 TI - High-level expression of porcine liver cytochrome P-450 reductase catalytic domain in Escherichia coli by modulating the predicted local secondary structure of mRNA. AB - A direct expression system for the solubilized catalytic domains of NADPH cytochrome P-450 reductase (sCPR) from rat (RsCPR) and porcine (PsCPR) in Escherichia coli cells was constructed using the expression plasmid pCWori(+). PsCPR was minimally expressed, whereas RsCPR was highly expressed. Replacement of the nucleotides encoding Thr(60)Ser(61)Ser(62) in PsCPR with those for Ala(60)Pro(61)Pro(62) in RsCPR markedly increased the expression level of the protein. The local secondary structures of the mRNAs, which were predicted with the prediction program GeneBee (http://www.genebee.msu.su), suggested that the intramolecular double strand between the ribosome binding site (RBS) and the Thr(60)Ser(61)Ser(62) codons in PsCPR, and/or the base-pairing at the initiation codon of the mRNAs significantly affected protein expression. Silent mutations were systematically introduced into the codons for Thr(58) and Thr(60)Ser(61) in PsCPR to modulate the local secondary structure of the mRNA. The expression level of the silently mutated PsCPR suggests that the expression level of PsCPR depends on the stability of the local structure at the RBS in the mRNA. A high-level expression system for wild-type PsCPR was constructed by introducing silent mutations at the codons for Thr(60)Ser(61) in PsCPR. The purified PsCPR showed the characteristic absorption spectral changes of sCPR after reduction with NADPH. The yield of purified PsCPR from 1 liter of culture fluid was 45.8 mg. These results substantiate that the introduction of silent mutations in the section of the gene encoding the N-terminal region of the protein based on the predicted local secondary structure of the mRNA at the RBS is a useful approach to control and increase the expression level of heterologous proteins in E. coli cells. PMID- 14561727 TI - Post-translational targeting of a tail-anchored green fluorescent protein to the endolpasmic reticulum. AB - Microsomal aldehyde dehydrogenase (msALDH) is a tail-anchored protein localized to the cytoplasmic face of the endoplasmic reticulum (ER). The carboxyl-terminal 35 amino acids of msALDH possess ER-targeting sequences in addition to a hydrophobic membrane-spanning domain. To study the mechanism for ER targeting of this protein in vivo, we took advantage of a green fluorescent protein-msALDH fusion protein containing the last 35 amino acids of msALDH [GFPALDH(35)]. When expressed from cDNA in COS-7 cells, the fusion protein was localized to the ER. We then prepared a recombinant fusion protein and injected it into the cytoplasm of COS-7 cells. The injected protein was correctly localized to the ER after a 30 min incubation at 37 degrees C. However, a recombinant fusion protein that contained only the transmembrane domain of msALDH failed to be targeted to the ER. When the assay was carried out at 4 degrees C, the recombinant GFPALDH(35) remained in the cytoplasm. Moreover, incubation of COS-7 cells under conditions of ATP depletion resulted in the cytoplasmic distribution of the injected protein. These results indicate that GFPALDH(35) is targeted to the ER post translationally via an ATP-dependent pathway. This microinjection system worked effectively in different mammalian cell types, suggesting a common mechanism for ER targeting of the tail-anchored protein. PMID- 14561728 TI - Structural elucidation of the protein- and membrane-binding properties of the N terminal tail domain of human annexin II. AB - The conformational preferences and the solution structure of AnxII(N31), a peptide corresponding to the full-length sequence (residues 1-31) of the human annexin II N-terminal tail domain, were investigated by circular dichroism (CD) and nuclear magnetic resonance (NMR) spectroscopy. CD results showed that AnxII(N31) adopts a mainly alpha-helical conformation in hydrophobic or membrane mimetic environments, while a predominantly random structure is adopted in aqueous buffer. In contrast to previous results of the annexin I N-terminal domain peptide [Yoon et al. (2000) FEBS Lett. 484, 241-245], calcium ions showed no effect on the structure of AnxII(N31). The NMR-derived structure of AnxII(N31) in 50% TFE/water mixture showed a horseshoe-like fold comprising the N-terminal amphipathic alpha-helix, the following loop, and the C-terminal helical region. Together, the results establish the first detailed structural data on the N terminal tail domain of annexin II, and suggest the possibility of the domain to undergo Ca(2+)-independent membrane-binding. PMID- 14561729 TI - The expression of proprotein convertase PACE4 is highly regulated by Hash-2 in placenta: possible role of placenta-specific basic helix-loop-helix transcription factor, human achaete-scute homologue-2. AB - PACE4 is a member of the mammalian subtilisin-like proprotein convertase (SPC) family, which contribute to the activation of transforming growth factor (TGF) beta family proteins. We previously reported that PACE4 is highly expressed in syncytiotrophoblasts of human placenta [Tsuji et al. (2003) BIOCHIM: Biophys. Acta 1645, 95-104]. In this study, the regulatory mechanism for PACE4 expression in placenta was analyzed using a human placental choriocarcinoma cell line, BeWo cells. Promoter analysis indicated that an E-box cluster (E4-E9) in the 5' flanking region of the PACE4 gene acts as a negative regulatory element. The binding of human achaete-scute homologue 2 (Hash-2) to the E-box cluster was shown by gel mobility-shift assay. The overexpression of Hash-2 caused a marked decrease in PACE4 gene expression. When BeWo cells were grown under low oxygen (2%) conditions, the expression of Hash-2 decreased, while that of PACE4 increased. In both cases, other SPCs, such as furin, PC5/6, and PC7/8, were not affected. Further, PACE4 expression was found to be developmentally regulated in rat placenta. By in situ hybridization, Mash-2 (mammalian achaete-scute homologue 2) mRNA was found to be expressed in the spongiotrophoblast layer where PACE4 was not expressed. In contrast, the PACE4 mRNA was expressed mainly in the labyrinthine layer where Mash-2 was not detected. These results suggest that PACE4 expression is down-regulated by Hash-2/Mash-2 in both human and rat placenta and that many bioactive proteins might be regulated by PACE4 activity. PMID- 14561730 TI - Role of p38 MAPK in lupeol-induced B16 2F2 mouse melanoma cell differentiation. AB - We examined the signaling mechanisms involved in the differentiation-inducing activity of lupeol toward B16 2F2 melanoma cells. alpha-Melanocyte stimulating hormone (alpha-MSH), forskolin and dibutyryl cAMP, which are believed to be cAMP elevating agents and analogues, enhanced lupeol-induced B16 2F2 cell differentiation. However, H89, an inhibitor of protein kinase A, completely abolished B16-2F2 cell differentiation induced by lupeol. Furthermore, we studied the role of mitogen-activated protein kinases (MAPKs) in lupeol-induced B16 2F2 cell differentiation. U0126, an inhibitor of MAPK kinases, induced B16 2F2 cell differentiation and enhanced the cell differentiation induced by lupeol. However, SB203580, a selective inhibitor of p38 MAPK, completely blocked lupeol-induced B16 2F2 cell differentiation. Western blot analysis revealed that 10 microM lupeol transiently elevated the level of phosphorylation of p38 MAPK. The phosphorylation of p38 MAPK was detected on the addition of 1 microM lupeone, another lupane triterpene, but was not induced by 1 microM lupeol. These results suggested that lupeol induced B16 2F2 cell differentiation through activation of p38 MAPK, and that the structural differences at C-3 of lupane triterpenes played an important role in the activation of p38 MAPK. PMID- 14561731 TI - Identification and characterization of a Xenopus homolog of Dbf4, a regulatory subunit of the Cdc7 protein kinase required for the initiation of DNA replication. AB - Dbf4 is a regulatory subunit for the Cdc7 protein kinase that is required for the initiation of eukaryotic DNA replication, but the precise roles of Dbf4-Cdc7 remain to be determined. Here we identified a Xenopus homolog of Dbf4 (XDbf4) and characterized XDbf4 and Xenopus Cdc7 (XCdc7) in Xenopus egg extracts. XDbf4 formed a complex with XCdc7 in egg extracts and activated XCdc7 kinase activity in vitro. In contrast with Dbf4 in yeast and mammalian cultured cells, the XDbf4 levels in egg extracts did not change during the cell cycle progression. XDbf4 was a phosphoprotein in interphase extracts, and was apparently hyperphosphorylated in cytostatic factor (CSF)-mediated, metaphase-arrested extracts and in mitotic extracts. However, the hyperphosphorylation of XDbf4 did not seem to affect the level of kinase activation, or chromatin binding of the XDbf4-XCdc7 complex. Upon release from CSF-arrest, XDbf4 was partially dephosphorylated and bound to chromatin. Interestingly, XDbf4 was loaded onto chromatin before XCdc7 during DNA replication in egg extracts. These results suggest that the function of XDbf4-XCdc7 during the early embryonic cell cycle is regulated in a manner distinct from that during the somatic cell cycle. PMID- 14561732 TI - Minimally modified LDL is an oxidized LDL enriched with oxidized phosphatidylcholines. AB - The oxidative modification of low-density lipoprotein (LDL) is involved in atherogenesis. Among a variety of modified LDLs mentioned in the literature, so called minimally modified LDL (MM-LDL) was reported to have pro-atherogenic properties despite minimal changes in its oxidative measures. After treatment of LDL with 1 micro M FeSO(4) at 4 degrees C for 96 h, the resulting MM-LDL showed a slight increase in thiobarbituric acid-reactive substances (TBARS) and little association with macrophages. On the other hand, heavily oxidized LDL, which was prepared by copper-induced oxidation of LDL at 37 degrees C, showed a sharp increase in TBARS and strong association with macrophages. By introducing a fluorometric procedure to detect aldehyde-containing phosphatidylcholines (aldehyde-PCs), we examined the amounts of aldehyde-PCs in modified LDL preparations. Aldehyde-PCs increased to 23.4 pmol/ microg protein in MM-LDL, which was more than four-fold higher than in the heavily oxidized LDL. We conclude that MM-LDL is a unique type of oxidized LDL enriched with aldehyde-PCs. PMID- 14561733 TI - Participation of proteasomes in Xenopus embryogenesis. AB - We examined the effects of various protease substrates on Xenopus laevis embryogenesis. Thirty-three peptidyl-MCA substrates were added to the culture medium in which Xenopus embryos were developing. Five of the 33 substrates were found to inhibit embryogenesis at the early gastrula stage or much earlier ones. These results suggest that proteases that hydrolyze these substrates are involved in embryonic development. We found that the developmental stage of embryos is crucial for these substrates to inhibit their development. We purified a protease that hydrolyzes Pyr-Arg-Thr-Lys-Arg-MCA, a substrate that inhibits embryogenesis, from Xenopus embryos. This protease turned out to be a component of proteasomes. We found that 4 of the 5 substrates that inhibit embryogenesis are among the proteasome substrates. Thus, we concluded that proteasomes play a crucial role in the development of Xenopus embryos. Possibly, various catalytic subunits in proteasomes function independently, in stage-specific manners. PMID- 14561734 TI - Cloning and analysis of the beta-lactamase gene from epsilon-poly-L-lysine producing actinomycete Streptomyces albulus IFO14147. AB - Streptomyces albulus IFO14147 produces epsilon-poly-L-lysine, which exhibits antimicrobial activity. It is necessary for its molecular breeding to develop host-vector systems. We recently found a novel cryptic plasmid, pNO33, in this strain. As part of a search for a selectable marker gene for pNO33, we report here the isolation and analysis of the beta-lactamase gene of this strain, which can grow on ampicillin-containing plates. It was shown that the beta-lactamase production in S. albulus was induced by ampicillin. By introducing a genomic library of S. albulus into Escherichia coli, a 3.6-kbp fragment was identified as the region involved in ampicillin resistance. It contained three open reading frames, all of which are highly homologous to the beta-lactamase (the blaL product) and its regulatory proteins (the blaA and blaB products) of S. cacaoi. The growth phenotypes and enzyme assaying of E. coli and S. lividans showed that the blaL homologue (blaSa) encodes a beta-lactamase required for ampicillin resistance. The beta-lactamae gene can be utilized as a selectable marker in a cloning vector of S. albulus. However, the beta-lactamase activity was decreased in E. coli and repressed in S. lividans by the blaA and blaB homologues (blaASa and blaBSa). It appears as if the blaASa product is a repressor of blaSa instead of an activator as in S. cacaoi. PMID- 14561735 TI - Evidence that HIV budding in primary macrophages occurs through the exosome release pathway. AB - Lipid rafts are specialized regions of cell membranes enriched in cholesterol and sphingolipids that are involved in immune activation and signaling. Studies in T cells indicate that these membrane domains serve as sites for release of human immunodeficiency virus (HIV). By budding through lipid rafts in T-cells, HIV selectively incorporates raft markers and excludes non-raft proteins. This process has been well studied in T-cells, but it is unknown whether lipid rafts serve as budding sites for HIV in macrophages. Recently, we proposed a new model of retroviral biogenesis called the Trojan exosome hypothesis (Gould, S. J., Booth, A., and Hildreth, J. E. K. (2003) Proc. Natl. Acad. Sci. U. S. A. 100, 10592-10597). This model proposes that retroviruses coopt the existing cellular machinery for exosomal release. Here, we performed the first test designed to differentiate between the lipid raft hypothesis of retroviral biogenesis and the Trojan exosome hypothesis. Using macrophages, we examined the relative abundance of several host proteins on the cell surface, in lipid rafts, and on both HIV particles and exosomes derived from these cells. Our results show significant differences in the abundance of host proteins on the cell surface and in HIV. Moreover, our data demonstrate discordance in the abundance of some proteins in lipid rafts and in HIV. Finally, our data reveal a strong concordance between the host cell protein profile of exosomes and that of HIV. These results strongly support the Trojan exosome hypothesis and its prediction that retroviral budding represents exploitation of a pre-existing cellular pathway of intercellular vesicle trafficking. PMID- 14561737 TI - Differential processing of CD4 T-cell epitopes from the protective antigen of Bacillus anthracis. AB - We have mapped CD4+ T-cell epitopes located in three domains of the recombinant protective antigen of Bacillus anthracis. Mouse T-cell hybridomas specific for these epitopes were generated to study the mechanisms of proteolytic processing of recombinant protective antigen for antigen presentation by bone marrow-derived macrophages. Overall, epitopes differed considerably in their processing requirements. In particular, the kinetics of presentation, ranging from 15 (fast) to 120 min (slow), suggested sequential liberation of epitopes during proteolytic processing of the intact PA molecule. Pretreatment of macrophages with ammonium chloride or inhibitors of the major enzyme families showed that T-cell responses to an epitope presented with fast kinetics were unaffected by raising endosomal pH or inhibiting cysteine or aspartic proteinases, suggesting presentation independent of lysosomal processing. In contrast, responses to epitopes presented with slower kinetics were dependent on low pH and the activity of cysteine or aspartic proteinases indicating a requirement for lysosomal processing. In addition, responses to all epitopes, whether their presentation was dependent on low pH or not, were prevented by treatment of macrophages with broad spectrum serine proteinase inhibitors. Thus, our data are consistent with a model of sequential antigen processing within the endosomal system, beginning with a pre processing step mediated by serine or metalloproteinases prior to further processing by lysosomal enzymes. Rapidly presented epitopes seemed to require only limited proteolysis at earlier stages of endocytosis, whereas the majority of epitopes required more extensive processing by neutral proteinases followed by lysosomal enzymes. PMID- 14561738 TI - Conformational variability of nucleo-cytoplasmic transport factors. AB - The transport of macromolecules between the nucleus and cytoplasm of eukaryotic cells is largely mediated by a single family of transport factors, the karyopherin or importin beta-like family. Structural and biochemical evidence suggests conformational flexibility of these modular HEAT-repeat proteins is crucial for their regulation. Here we use small angle x-ray scattering to assess the extent of conformational variation within a set of nuclear import and export factors. The study reveals that importin beta, transportin, and the exportin Xpo t share a similar S-like superhelical conformation in their unbound state. There are no obvious differences in the overall structures that might generally distinguish nuclear export from nuclear import mediators. Two other members of the family, the exportins Cse1 and Xpo1, possess a significantly more globular conformation, indicating that the extended S-like architecture is not a hallmark of all karyopherins. Binding of RanGTP/cargo to importin beta, transportin, and Xpo-t triggers distinct conformational responses, suggesting that even closely related karyopherins employ different mechanisms of conformational regulation and that cargo and nuclear pore-interacting surfaces of the different receptors may be unique. PMID- 14561736 TI - Regulation of tension-induced mechanotranscriptional signals by the microtubule network in fibroblasts. AB - Mechanical loading of connective tissues induces the expression of extracellular matrix and cytoskeletal genes that are involved in matrix remodeling. These processes depend in part on force transmission through beta1 integrins and actin filaments, but the role of microtubules in regulating mechanotranscriptional responses is not well defined. We assessed the involvement of microtubules in the mechanotranscriptional regulation of filamin A, an actin-cross-linking protein that protects cells against force-induced apoptosis by stabilizing cell membranes. Collagen-coated magnetite beads and magnetic fields were used to apply tensile forces to cultured fibroblasts at focal adhesions. Force enhanced recruitment of alpha-tubulin and the plus end microtubule-binding protein cytoplasmic linker protein-170 (CLIP-170) at focal adhesions. Immunoprecipitation studies demonstrated no direct binding of tubulin to actin or filamin A, but CLIP 170 interacted with tubulin, filamin A, and beta-actin. The association of CLIP 170 with beta-actin was enhanced by force. Force activated the p38 mitogen activated protein kinase, increased filamin A expression, and induced the relocation of p38 and filamin A to focal adhesions. Disruption of microtubules with nocodazole, independent of force application, enhanced filamin A expression and Sp1-mediated filamin A promoter activity, while stabilization of microtubules with Taxol inhibited force induction of both filamin A mRNA and protein. We conclude that in response to tensile forces applied through beta1 integrins and actin the microtubule network modulates mechanotranscriptional coupling of filamin A. PMID- 14561739 TI - Bile acids up-regulate death receptor 5/TRAIL-receptor 2 expression via a c-Jun N terminal kinase-dependent pathway involving Sp1. AB - Bile acids up-regulate death receptor 5 (DR5)/TRAIL-receptor 2 (TRAIL-R2) expression thereby sensitizing hepatocytes to TRAIL-mediated apoptosis. However, the precise mechanism by which bile acids enhance DR5/TRAIL-R2 expression is unknown. Although several bile acids enhanced DR5/TRAIL-R2 expression, deoxycholic acid (DCA) was the most potent. DCA stimulated JNK activation and the JNK inhibitor SP600125 blocked DCA-induced DR5/TRAIL-R2 mRNA and protein expression. Reporter gene analysis identified a 5'-flanking region containing two Sp1 binding sites within the DR5/TRAIL-R2 promoter as bile acid responsive. Sp1 binding to one of the two sites was enhanced by DCA treatment as evaluated by electrophoretic mobility shift assays and chromatin immunoprecipitation studies. JNK inhibition with SP600125 also blocked binding of Sp1 to the DR5/TRAIL-R2 promoter. Finally, point mutations of the Sp1 binding site attenuated promoter activity. In conclusion, Sp1 is a bile acid-responsive transcription factor that mediates DR5/TRAIL-R2 gene expression downstream of JNK. PMID- 14561740 TI - A chromatin immunoprecipitation screen reveals protein kinase Cbeta as a direct RUNX1 target gene. AB - RUNX1 (also known as AML1) is a DNA-binding transcription factor that functions as a tumor suppressor and developmental determinant in hematopoietic cells. Target promoters have been identified primarily through the use of differential expression strategies and candidate gene approaches but not biochemical screens. Using a chromatin immunoprecipitation screen, we identified protein kinase Cbeta as a direct RUNX1 target gene and demonstrate that endogenous RUNX1 binds the chromatinized protein kinase Cbeta promoter of U937 cells. A phylogenetically conserved RUNX1-binding site within the PKCbeta promoter binds RUNX1 in electrophoretic mobility shift analyses and confers RUNX1 responsiveness on a heterologous promoter. Changes in RUNX1 activity affect endogenous protein kinase Cbeta expression, and a dominant-negative form of RUNX1 protects U937 cells from apoptotic stimuli previously shown to be dependent on protein kinase Cbeta. This protection can be reversed by the ectopic expression of protein kinase Cbeta. Together these findings demonstrate that protein kinase Cbeta is a direct, downstream target of RUNX1 and links RUNX1 to a myeloid apoptotic pathway. PMID- 14561741 TI - Residues in internal repeats of the rice cation/H+ exchanger are involved in the transport and selection of cations. AB - In plants, the cation/H+ exchanger (CAX) translocates Ca2+ and other metal ions into vacuoles using the H+ gradient formed by H+-ATPase and H+-pyrophosphatase. Such exchangers carrying 11 transmembrane domains (TMs) have been isolated from plants, yeast, and bacteria. In this study, multiple sequence alignment of several CAXs revealed the presence of highly conserved 36-residue regions between TM3 and TM4 and between TM8 and TM9. These two repetitive motifs are designated repeats c-1 and c-2. Using site-directed mutagenesis, we generated 31 mutations in the repeats of the Oryza sativa CAX, which translocates Ca2+ and Mn2+. Mutant exchangers were expressed in a Saccharomyces cerevisiae strain that is sensitive to Ca2+ and Mn2+ because of the absence of vacuolar Ca2+-ATPase and the Ca2+/H+ exchanger. Mutant exchangers were classified into six classes according to their tolerance for Ca2+ and Mn2+. For example, the class III mutants had no tolerance for either ion, and the class IV mutants had tolerance only for Ca2+. The biochemical function of each residue was estimated. We investigated the membrane topology of the repeats using a method combining cysteine mutagenesis and sulfhydryl reagents. Our results suggest that repeat c-1 re-enters the membrane from the vacuolar luminal side and forms a solution-accessible region. Furthermore, several residues in repeats c-1 and c-2 were found to be conserved in animal Na+/Ca2+ exchangers. Finally, we suggest that these re-entrant repeats may form a vestibule or filter for cation selection. PMID- 14561742 TI - Isoform-specific phosphorylation of metabotropic glutamate receptor 5 by protein kinase C (PKC) blocks Ca2+ oscillation and oscillatory translocation of Ca2+ dependent PKC. AB - Prolonged activation of metabotropic glutamate receptor 5a (mGluR5a) causes synchronized oscillations in intracellular calcium, inositol 1,4,5-trisphosphate production, and protein kinase C (PKC) activation. Additionally, mGluR5 stimulation elicited cyclical translocations of myristoylated alanine-rich protein kinase C substrate, which were opposite to that of gammaPKC (i.e. from plasma membrane to cytosol) and dependent on PKC activity, indicating that myristoylated alanine-rich protein kinase C substrate is repetitively phosphorylated by oscillating gammaPKC on the plasma membrane. Mutation of mGluR5 Thr(840) to aspartate abolished the oscillation of gammaPKC, but the mutation to alanine (T840A) did not. Cotransfection of gammaPKC with betaIIPKC, another Ca2+ dependent PKC, resulted in synchronous oscillatory translocation of both classical PKCs. In contrast, cotransfection of deltaPKC, a Ca2+-independent PKC, abolished the oscillations of both gammaPKC and inositol 1,4,5-trisphosphate. Regulation of the oscillations was dependent on deltaPKC kinase activity but not on gammaPKC. Furthermore, the T840A-mGluR5-mediated oscillations were not blocked by the deltaPKC overexpression. These results revealed that activation of mGluR5 causes translocation of both gammaPKC and deltaPKC to the plasma membrane. deltaPKC, but not gammaPKC, phosphorylates mGluR5 Thr(840), leading to the blockade of both Ca2+ oscillations and gammaPKC cycling. This subtype-specific targeting proposes the molecular basis of the multiple functions of PKC. PMID- 14561743 TI - Lec3 Chinese hamster ovary mutants lack UDP-N-acetylglucosamine 2-epimerase activity because of mutations in the epimerase domain of the Gne gene. AB - Lec3 Chinese hamster ovary (CHO) cell glycosylation mutants have a defect in sialic acid biosynthesis that is shown here to be reflected most sensitively in reduced polysialic acid (PSA) on neural cell adhesion molecules. To identify the genetic origin of the phenotype, genes encoding different factors required for sialic acid biosynthesis were transfected into Lec3 cells. Only a Gne cDNA encoding UDP-GlcNAc 2-epimerase:ManNAc kinase rescued PSA synthesis. In an in vitro UDP-GlcNAc 2-epimerase assay, Lec3 cells had no detectable UDP-GlcNAc 2 epimerase activity, and Lec3 cells grown in serum-free medium were essentially devoid of sialic acid on glycoproteins. The Lec3 phenotype was rescued by exogenously added N-acetylmannosamine or mannosamine but not by the same concentrations of N-acetylglucosamine, glucosamine, glucose, or mannose. Sequencing of CHO Gne cDNAs identified a nonsense (E35stop) and a missense (G135E) mutation, respectively, in two independent Lec3 mutants. The G135E Lec3 mutant transfected with a rat Gne cDNA had restored in vitro UDP-GlcNAc 2 epimerase activity and cell surface PSA expression. Both Lec3 mutants were similarly rescued with a CHO Gne cDNA and with CHO Gne encoding the known kinase deficient D413K mutation. However, cDNAs encoding the known epimerase-deficient mutation H132A or the new Lec3 G135E Gne mutation did not rescue the Lec3 phenotype. The G135E Gne missense mutation is a novel mechanism for inactivating UDP-GlcNAc 2-epimerase activity. Lec3 mutants with no UDP-GlcNAc 2-epimerase activity represent sensitive hosts for characterizing disease-causing mutations in the human GNE gene that give rise to sialuria, hereditary inclusion body myopathy, and Nonaka myopathy. PMID- 14561744 TI - Histone H2AX phosphorylation as a predictor of radiosensitivity and target for radiotherapy. AB - Based on the role of phosphorylation of the histone H2A variant H2AX in recruitment of DNA repair and checkpoint proteins to the sites of DNA damage, we have investigated gammaH2AX as a reporter of tumor radiosensitivity and a potential target to enhance the effectiveness of radiation therapy. Clinically relevant ionizing radiation (IR) doses induced similar patterns of gammaH2AX focus formation or immunoreactivity in radiosensitive and radioresistant human tumor cell lines and xenografted tumors. However, radiosensitive tumor cells and xenografts retained gammaH2AX for a greater duration than radioresistant cells and tumors. These results suggest that persistence of gammaH2AX after IR may predict tumor response to radiotherapy. We synthesized peptide mimics of the H2AX carboxyl-terminal tail to test whether antagonizing H2AX function affects tumor cell survival following IR. The peptides did not alter the viability of unirradiated tumor cells, but both blocked induction of gammaH2AX foci by IR and enhanced cell death in irradiated radioresistant tumor cells. These results suggest that H2AX is a potential molecular target to enhance the effects of radiotherapy. PMID- 14561745 TI - Distinct RE-1 silencing transcription factor-containing complexes interact with different target genes. AB - Establishment of neuronal identity requires co-ordinated expression of specific batteries of genes. These programs of gene expression are executed by activation of neuron-specific genes in neuronal cells and their repression in non-neuronal cells. Such co-ordinate regulation requires that individual activators and repressors regulate transcription from specific subsets of their potential target genes, yet we know little of the mechanisms that underlie this selective process. The RE-1 silencing transcription factor (REST) is a repressor that is proposed to silence transcription of numerous neuron-specific genes in non-neuronal cells via recruitment of two independent histone deacetylase (HDAC)-containing co-repressor complexes. However, in vivo, REST appears to be an obligate silencer for only a minority of RE-1-bearing genes. Here we examine the interaction of REST, Co-REST, Sin3A, HDAC1, and HDAC2 with two archetypical endogenous target genes, the M4 muscarinic receptor and the sodium type II channel (NaV1.2) genes. We find that these genes are present in distinct chromosomal domains. The NaV1.2 gene is actively transcribed but repressed by REST independently of histone deacetylation or DNA methylation and does not co-localize with epigenetic markers of silence, including dimethylation of H3K9 and HP1. In contrast, the M4 gene is maintained in a silent state independently of REST and co-localizes with dimethylated H3K9 and HP1alpha and HP1gamma, characteristic of silenced or senescent euchromatic DNA. This contrasts with the coordinate REST-dependent regulation of this locus reported previously. Taken together, we infer that distinct repressor complexes and mechanisms are operative at particular loci even in cell lines derived from a common embryological origin. PMID- 14561746 TI - Molecular modeling correctly predicts the functional importance of Phe594 in transmembrane helix 11 of the multidrug resistance protein, MRP1 (ABCC1). AB - The human ATP-binding cassette (ABC) transporter, multidrug resistance protein 1 (MRP1/ABCC1), confers resistance to a broad range of anti-cancer agents and transports a variety of organic anions. At present, essentially no structural data exists for MRP1 that might be used to elucidate its mechanism of transport. Consequently, we have applied a modeling strategy incorporating crystal and indirect structural data from other ABC transporters to construct a model of the transmembrane domains of the core region of MRP1 that includes the amino acid side chains. Three conserved Trp residues and one non-conserved Tyr residue, shown previously to be of functional importance (Koike, K., Oleschuk, C. J., Haimeur, A., Olsen, S. L., Deeley, R. G., and Cole, S. P. C. (2002) J. Biol. Chem. 277, 49495-49503), were found to line the "pore" in our model proximal to the membrane cytosol interface. A fifth aromatic residue (Phe594) was identified that, with the Trp and Tyr residues, completed a ring or "basket" of aromatic amino acids and, accordingly, we postulated that it would also be of functional importance. To test this idea, MRP1-Phe594 mutants were expressed in human embryonic kidney cells, and their properties were examined using membrane vesicles. Substitution of Phe594 with Ala substantially reduced or eliminated the transport of five organic anion substrates by MRP1 and abrogated the binding of leukotriene C4. On the other hand, the conservatively substituted F594W and F594Y mutants remained transport competent, although significant substrate- and substitution-specific changes were observed. These studies provide some structural insight into a possible substrate binding/transport site of MRP1 at the beginning of a putative substrate translocation pathway and demonstrate the usefulness of modeling for directing structure-function analyses of this transporter. PMID- 14561747 TI - The mitochondrial citrate transport protein: probing the secondary structure of transmembrane domain III, identification of residues that likely comprise a portion of the citrate transport pathway, and development of a model for the putative TMDIII-TMDIII' interface. AB - The mitochondrial citrate transport protein (CTP) has been investigated by mutating 28 consecutive residues within transmembrane domain III (TMDIII), one at a time, to cysteine. A cysteine-less CTP that retains wild-type functional properties, served as the starting template. The single Cys CTP mutants were abundantly expressed in Escherichia coli, isolated, and functionally reconstituted in a liposomal system. The accessibility of each single Cys mutant to two methanethiosulfonate reagents was evaluated by determining the rate constants for inhibition of CTP function. These rate constants varied by over five orders of magnitude. With two independent data sets we observed peaks and troughs in the rate constant data at identical amino acid positions and a periodicity of 4 was observed from residues 123-137. Based on the pattern of accessibility we conclude that residues 123-137 exist as an alpha-helix. Although less certain, a combination of the rate constant data and the specific activity data with the single Cys mutants suggests that the alpha-helical secondary structure may extend to residue 113. Furthermore, the rate constant data define water-accessible and water-inaccessible faces of the helix. We infer that the water-accessible face comprises a portion of the substrate translocation pathway through the CTP, whereas the water-inaccessible surface faces the lipid bilayer. Finally, based on a combination of the CTP inhibition rate constant data and the existence of significant sequence identity with a transmembrane segment within glycophorin A that forms a portion of its dimer interface, a model for a putative CTP TMDIII-TMDIII' dimer interface has been developed. PMID- 14561748 TI - Biosynthesis, purification, and substrate specificity of severe acute respiratory syndrome coronavirus 3C-like proteinase. AB - The 3C-like proteinase of severe acute respiratory syndrome (SARS) coronavirus has been proposed to be a key target for structural-based drug design against SARS. In order to understand the active form and the substrate specificity of the enzyme, we have cloned, expressed, and purified SARS 3C-like proteinase. Analytic gel filtration shows a mixture of monomer and dimer at a protein concentration of 4 mg/ml and mostly monomer at 0.2 mg/ml, which correspond to the concentration used in the enzyme assays. The linear decrease of the enzymatic-specific activity with the decrease of enzyme concentration revealed that only the dimeric form is active and the dimeric interface could be targeted for structural-based drug design against SARS 3C-like proteinase. By using a high pressure liquid chromatography assay, SARS 3C-like proteinase was shown to cut the 11 peptides covering all of the 11 cleavage sites on the viral polyprotein with different efficiency. The two peptides corresponding to the two self-cleavage sites are the two with highest cleavage efficiency, whereas peptides with non-canonical residues at P2 or P1' positions react slower. The P2 position of the substrates seems to favor large hydrophobic residues. Secondary structure studies for the peptide substrates revealed that substrates with more beta-sheetlike structure tend to react fast. This study provides a basic understanding of the enzyme catalysis and a full substrate specificity spectrum for SARS 3C-like proteinase, which are helpful for structural-based inhibitor design against SARS and other coronavirus. PMID- 14561749 TI - PTEN M-CBR3, a versatile and selective regulator of inositol 1,3,4,5,6 pentakisphosphate (Ins(1,3,4,5,6)P5). Evidence for Ins(1,3,4,5,6)P5 as a proliferative signal. AB - The PTEN (phosphatase and tensin homologue deleted on chromosome 10) tumor suppressor is a phosphatidylinositol 3,4,5-trisphosphate (PtdInsP3) 3-phosphatase that plays a crucial role in regulating many cellular processes by antagonizing the phosphoinositide 3-kinase signaling pathway. Although able to metabolize soluble inositol phosphates in vitro, the question of their significance as physiological substrates is unresolved. We show that inositol phosphates are not regulated by wild type PTEN, but that a synthetic mutant, PTEN M-CBR3, previously thought to be inactive toward inositides, can selectively regulate inositol 1,3,4,5,6-pentakisphosphate (Ins(1,3,4,5,6)P5). Transfection of U87-MG cells with PTEN M-CBR3 lowered Ins(1,3,4,5,6)P5 levels by 60% without detectable effect on PtdInsP3. Although PTEN M-CBR3 is a 3-phosphatase, levels of myo-inositol 1,4,5,6 tetrakisphosphate were not increased, whereas myo-inositol 1,3,4,6 tetrakisphospate levels increased by 80%. We have used PTEN M-CBR3 to study the physiological function of Ins(1,3,4,5,6)P5 and have found that Ins(1,3,4,5,6)P5 does not modulate PKB phosphorylation, nor does it regulate clathrin-mediated epidermal growth factor receptor internalization. By contrast, PTEN M-CBR3 expression, and the subsequent lowering of Ins(1,3,4,5,6)P5, are associated with reduced anchorage-independent colony formation and anchorage-dependent proliferation in U87-MG cells. Our results, together with previously published data, suggest that Ins(1,3,4,5,6)P5 has a role in proliferation. PMID- 14561750 TI - Phospholipase C-independent activation of glycogen synthase kinase-3beta and C terminal Src kinase by Galphaq. AB - It is generally thought that activation of phospholipase Cbeta (PLCbeta) by Galphaq accounts for most of the effects of Gq-coupled receptors. Here we describe a novel effect of Galphaq that is independent of the PLCbeta pathway. Expression of the constitutively active Galphaq mutant Galphaq(Q209L) promoted an increase in glycogen synthase kinase-3beta (GSK-3beta) activity that was associated with increased phosphorylation of Tyr216 on GSK-3beta. Galphaq(Q209L) AA, a mutant that cannot activate PLCbeta, also induced GSK-3beta activation and phosphorylation of Tyr216. We speculate that the protein-tyrosine kinase Csk (C terminal Src kinase), which is also activated by Galphaq(Q209L) and Galphaq(Q209L)-AA, acts upstream of GSK-3beta. Expression of Csk accentuated the activation of GSK-3beta by Galphaq(Q209L), whereas catalytically inactive Csk blocked GSK-3beta activation by Galphaq(Q209L). Recombinant Csk phosphorylated and activated GSK-3beta in vitro, and GSK-3beta coprecipitated with Csk from cell lysates. These results suggest that activation of Csk and GSK-3beta by Galphaq may contribute to the physiological and pathological effects of Gq-coupled receptors. PMID- 14561751 TI - A subfamily of acidic alpha-K(+) toxins. AB - Three homologous acidic peptides have been isolated from the venom of three different Parabuthus scorpion species, P. transvaalicus, P. villosus, and P. granulatus. Analysis of the primary sequences reveals that they structurally belong to subfamily 11 of short chain alpha-K(+)-blocking peptides (Tytgat, J., Chandy, K. G., Garcia, M. L., Gutman, G. A., Martin-Eauclaire, M. F., van der Walt, J. J., and Possani, L. D. (1999) Trends Pharmacol. Sci. 20, 444-447). These toxins are 36-37 amino acids in length and have six aligned cysteine residues, but they differ substantially from the other alpha-K(+) toxins because of the absence of the critical Lys(27) and their total overall negative charge. Parabutoxin 1 (PBTx1), which has been expressed by recombinant methods, has been submitted to functional characterization. Despite the lack of the Lys(27), this toxin blocks several Kv1-type channels heterologously expressed in Xenopus oocytes but with low affinities (micromolar range). Because a relationship between the biological activity and the acidic residue substitutions may exist, we set out to elucidate the relative impact of the acidic character of the toxin and the lack of the critical Lys(27) on the weak activity of PBTx1 toward Kv1 channels. To achieve this, a specific mutant named rPBTx1 T24F/V26K was made recombinantly and fully characterized on Kv1-type channels heterologously expressed in Xenopus oocytes. Analysis of rPBTx1 T24F/V26K displaying an affinity toward Kv1.2 and Kv1.3 channels in the nanomolar range shows the importance of the functional dyad above the acidic character of this toxin. PMID- 14561752 TI - N-acetylglucosaminyltransferase V expression levels regulate cadherin-associated homotypic cell-cell adhesion and intracellular signaling pathways. AB - A common glycan alteration in transformed cells and human tumors is the highly elevated levels of N-linked beta(1,6)glycans caused by increased transcription of N-acetylglucosaminyltransferase V (GnT-V). Here, we define the involvement of GnT V in modulation of homotypic cell-cell adhesion in human fibrosarcoma HT1080 and mouse NIH3T3 cells. Increased GnT-V expression resulted in a significant decrease in the rates of calcium-dependent cell-cell adhesion. Reduced cell-cell adhesion was blocked by function-blocking antibody against N-cadherin and abrogated by pre treatment of cells with swainsonine, demonstrating the involvement of N-cadherin in the cell-cell adhesion and that changes in N-linked beta(1,6)glycan expression are responsible for the reduction in rates of adhesion, although this reduction could be mediated by the altered N-linked glycosylation of glycoproteins other than N-cadherin. Overexpression of GnT-V had no effect on the levels of cell surface expression of N-cadherin; however, it did cause a marked enhancement of both beta(1,6) branching and poly-N-acetyllactosamine expression on N-cadherin. GnT-V overexpression resulted in decreased N-cadherin clustering on the cell surface induced by anti-N-cadherin antibody and affected the outside-in signal transduction pathway of ERK mediated by N-cadherin. Overexpression of GnT-V sensitized stimulation of tyrosine phosphorylation of catenins by growth factors and expression of v-src, which is consistent with its reduction of cell-cell adhesion. In vitro, GnT-V-overexpressing cells showed increased motility concomitant with increased phosphorylation of catenins. Moreover, GnT-V-deficient embryo fibroblasts from GnT-V homozygous null mice (GnT-V(-/-)) express N cadherin and showed significantly increased levels of N-cadherin-based cell-cell adhesion compared with those from GnT-V(+/-) mice. These levels of adhesion were inhibited significantly by transient expression of GnT-V, confirming the hypothesis that levels of GnT-V can regulate cadherin-associated homotypic cell cell adhesion. Aberrant N-linked beta(1,6) branching that occurs during oncogenesis can, therefore, lessen cell-cell adhesion, contributing to increased cellular motility and invasiveness. PMID- 14561753 TI - Intra-Golgi protein transport depends on a cholesterol balance in the lipid membrane. AB - Transport of proteins between intracellular membrane compartments is mediated by a protein machinery that regulates the budding and fusion processes of individual transport steps. Although the core proteins of both processes are defined at great detail, much less is known about the involvement of lipids. Here we report that changing the cellular balance of cholesterol resulted in changes of the morphology of the Golgi apparatus, accompanied by an inhibition of protein transport. By using a well characterized cell-free intra-Golgi transport assay, these observations were further investigated, and it was found that the transport reaction is sensitive to small changes in the cholesterol content of Golgi membranes. Addition as well as removal of cholesterol (10 +/- 6%) to Golgi membranes by use of methyl-beta-cyclodextrin specifically inhibited the intra Golgi transport assay. Transport inhibition occurred at the fusion step. Modulation of the cholesterol content changed the lipid raft partitioning of phosphatidylcholine and heterotrimeric G proteins, but not of other (non) lipid raft proteins and lipids. We suggest that the cholesterol balance in Golgi membranes plays an essential role in intra-Golgi protein transport and needs to be carefully regulated to maintain the structural and functional organization of the Golgi apparatus. PMID- 14561754 TI - Molecular components of a cell death pathway activated by endoplasmic reticulum stress. AB - Alterations in Ca2+ homeostasis and accumulation of misfolded proteins in the endoplasmic reticulum (ER) cause ER stress that ultimately leads to programmed cell death. Recent studies have shown that ER stress triggers programmed cell death via an alternative intrinsic pathway of apoptosis that, unlike the intrinsic pathway described previously, is independent of Apaf-1 and cytochrome c. In the present work, we have used a set of complementary approaches, including two-dimensional gel electrophoresis coupled with matrix-assisted laser desorption ionization-time-of-flight mass spectrometry and nano-liquid chromatography electrospray ionization mass spectrometry with tandem mass spectrometry, RNA interference, co-immunoprecipitation, immunodepletion of candidate proteins, and reconstitution studies, to identify mediators of the ER stress-induced cell death pathway. Our data identify two molecules, valosin-containing protein and apoptosis-linked gene-2 (ALG-2), that appear to play a role in mediating ER stress-induced cell death. PMID- 14561755 TI - A corresponding tyrosine residue in the C2/factor B type A domain is a hot spot in the decay acceleration of the complement C3 convertases. AB - The cleavage of C3 by the C3 convertases (C3bBb and C4b2a) determines whether complement activation proceeds. Dissociation (decay acceleration) of these central enzymes by the regulators decay-accelerating factor (DAF), complement receptor 1 (CR1), factor H, and C4-binding protein (C4BP) controls their function. In a previous investigation, we obtained evidence implicating the alpha4/5 region of the type A domain of Bb (especially Tyr338) in decay acceleration of C3bBb and proposed this site as a potential interaction point with DAF and long homologous repeat A of CR1. Because portions of only two DAF complement control protein domains (CCPs), CCP2 and CCP3, are necessary to mediate its decay of the CP C3 convertase (as opposed to portions of at least three CCPs in all other cases, e.g. CCPs 1-3 of CR1), DAF/C4b2a provides the simplest structural model for this reaction. Therefore, we examined the importance of the C2 alpha4/5 site on decay acceleration of C4b2a. Functional C4b2a complexes made with the C2 Y327A mutant, the C2 homolog to factor B Y338A, were highly resistant to DAF, C4BP, and long homologous repeat A of CR1, whereas C2 substitutions in two nearby residues (N324A and L328A) resulted in partial resistance. Our new findings indicate that the alpha4/5 region of C2a is critical to decay acceleration mediated by DAF, C4BP, and CR1 and suggest that decay acceleration of C4b2a and C3bBb requires interaction of the convertase alpha4/5 region with a CCP2/CCP3 site of DAF or structurally homologous sites of CR1 and C4BP. PMID- 14561756 TI - The caveolin scaffolding domain modifies 2-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor binding properties by inhibiting phospholipase A2 activity. AB - Activation of the enzyme phospholipase (PLA 2) has been proposed to be part of the molecular mechanism involved in the alteration of 2-amino-3-hydroxy-5-methyl 4-isoxazole propionate (AMPA) glutamate receptor responsiveness during long term changes in synaptic plasticity (long term potentiation). This study assesses the effect of the caveolin-1 scaffolding domain (CSD) on the activity of the regulatory enzyme PLA2. Caveolin-1 is a 22-kDa cholesterol-binding membrane protein known to inhibit the activity of most of its interacting partners. Our results show that the calcium-dependent cytosolic form of PLA2 (cPLA2) and caveolin-1 co-localized in mouse primary hippocampal neuron cultures and that they were co-immunoprecipitated from mouse hippocampal homogenates. A peptide corresponding to the scaffolding domain of caveolin-1 (Cav-(82-101)) dramatically inhibited cPLA2 activity in purified hippocampal synaptoneurosomes. Activation of endogenous PLA2 activity with KCl or melittin increased the binding of [3H]AMPA to its receptor. This effect was almost completely abolished by the addition of the CSD peptide to these preparations. Moreover, we demonstrated that the inhibitory action of the CSD peptide on AMPA receptor binding properties is specific (because a scrambled version of this peptide failed to have any effect) and that it is mediated by an inhibition of PLA2 enzymatic activity (because the CSD peptide failed to have an effect in membrane preparations lacking endogenous PLA2 activity). These results raised the possibility that caveolin-1, via the inhibition of cPLA2 enzymatic activity, may interfere with synaptic facilitation and long term potentiation formation in the hippocampus. PMID- 14561757 TI - Structural elements contribute to the calcium/calmodulin dependence on enzyme activation in human endothelial nitric-oxide synthase. AB - Two regions, located at residues 594-606/614-645 and residues 1165-1178, are present in the reductase domain of human endothelial nitric-oxide synthase (eNOS) but absent in its counterpart, inducible nitric-oxide synthase (iNOS). We previously demonstrated that removing residues 594-606/614-645 resulted in an enzyme (Delta45) containing an intrinsic calmodulin (CaM) purified from an Sf9/baculovirus expression system (Chen, P.-F., and Wu, K.K. (2000) J. Biol. Chem. 275, 13155-13163). Here we have further elucidated the differential requirement of Ca2+/CaM for enzyme activation between eNOS and iNOS by either deletion of residues 1165-1178 (Delta14) or combined deletions of residues 594 606/614-645 and 1165-1178 (Delta45/ Delta14) from eNOS to mimic iNOS. We measured the catalytic rates using purified proteins completely free of CaM. Steady-state analysis indicated that the Delta45 supported NO synthesis in the absence of CaM at 60% of the rate in its presence, consistent with our prior result that CaM bound Delta45 retained 60% of its activity in the presence of 10 mm EGTA. Mutant Delta14 displayed a 1.5-fold reduction of EC50 for Ca2+/CaM-dependence in l citrulline formation, and a 2-4-fold increase in the rates of NO synthesis, NADPH oxidation, and cytochrome c reduction relative to the wild type. The basal rates of double mutant Delta45/Delta14 in NO production, NADPH oxidation, and cytochrome c reduction were 3-fold greater than those of CaM-stimulated wild-type eNOS. Interestingly, all three activities of Delta45/ Delta14 were suppressed rather than enhanced by Ca2+/CaM, indicating a complete Ca2+/CaM independence for those reactions. The results suggest that the Ca2+/CaM-dependent catalytic activity of eNOS appears to be conferred mainly by these two structural elements, and the interdomain electron transfer from reductase to oxygenase domain does not require Ca2+/CaM when eNOS lacks these two segments. PMID- 14561758 TI - The secreted frizzled related protein 2 (SFRP2) gene is a target of the Pax2 transcription factor. AB - Despite their essential role in vertebrate development, the function of Pax proteins in gene regulation is not well understood. To identify potential genes regulated by the Pax2 protein, we screened embryonic kidney cells transformed with Pax2-expressing retroviruses for genes activated in response to Pax2 expression. In this system, the gene encoding the secreted frizzled related protein, Sfrp2, was strongly activated in all Pax2b-expressing cells. This activation of Sfrp2 expression correlated with changes in chromatin structure at the Sfrp2 locus, particularly in and around regions of Pax2 binding. Although the amount of Pax2-dependent transactivation was low in transient assays, the data suggests that local alterations of chromatin structure by Pax proteins can greatly enhance expression when presented in the right cellular context. PMID- 14561759 TI - Proteomic analysis of rat liver peroxisome: presence of peroxisome-specific isozyme of Lon protease. AB - Subcellular proteomics, which includes isolation of subcellular components prior to a proteomic analysis, is advantageous not only in characterizing large macro molecular complexes such as organelles but also in elucidating mechanisms of protein transport and organelle biosynthesis. Because of the high sensitivity achieved by the present proteomics technology, the purity of samples to be analyzed is important for the interpretation of the results obtained. In the present study, peroxisomes isolated from rat liver by usual cell fractionation were further purified by immunoisolation using a specific antibody raised against a peroxisomal membrane protein, PMP70. The isolated peroxisomes were analyzed by SDS-PAGE combined with liquid chromatography/mass spectrometry. Altogether 34 known peroxisomal proteins were identified in addition to several mitochondrial and microsomal proteins. Some of the latter may reside in the peroxisomes as well. Analysis of membrane fractions identified all known peroxins except for Pex7. Two new peroxisomal proteins of unknown function were of high abundance. One is a bi-functional protein consisting of an aminoglycoside phosphotransferase domain and an acyl-CoA dehydrogenase domain. The other is a newly identified peroxisome-specific isoform of Lon protease, an ATP-dependent protease with chaperone-like activity. The peroxisomal localization of the protein was confirmed by immunological techniques. The peroxisome-type Lon protease, which is distinct from the mitochondrial isoform, may play an important role in the peroxisomal biogenesis. PMID- 14561760 TI - Photoreceptor cGMP phosphodiesterase delta subunit (PDEdelta) functions as a prenyl-binding protein. AB - Bovine PDEdelta was originally copurified with rod cGMP phosphodiesterase (PDE) and shown to interact with prenylated, carboxymethylated C-terminal Cys residues. Other studies showed that PDEdelta can interact with several small GTPases including Rab13, Ras, Rap, and Rho6, all of which are prenylated, as well as the N-terminal portion of retinitis pigmentosa GTPase regulator and Arl2/Arl3, which are not prenylated. We show by immunocytochemistry with a PDEdelta-specific antibody that PDEdelta is present in rods and cones. We find by yeast two-hybrid screening with a PDEdelta bait that it can interact with farnesylated rhodopsin kinase (GRK1) and that prenylation is essential for this interaction. In vitro binding assays indicate that both recombinant farnesylated GRK1 and geranylgeranylated GRK7 co-precipitate with a glutathione S-transferase-PDEdelta fusion protein. Using fluorescence resonance energy transfer techniques exploiting the intrinsic tryptophan fluorescence of PDEdelta and dansylated prenyl cysteines as fluorescent ligands, we show that PDEdelta specifically binds geranylgeranyl and farnesyl moieties with a Kd of 19.06 and 0.70 microm, respectively. Our experiments establish that PDEdelta functions as a prenyl binding protein interacting with multiple prenylated proteins. PMID- 14561761 TI - Facilitated drug influx by an energy-uncoupled secondary multidrug transporter. AB - The majority of bacterial multidrug resistance transporters belong to the class of secondary transporters. LmrP is a proton/drug antiporter of Lactococcus lactis that extrudes positively charged lipophilic substrates from the inner leaflet of the membrane to the external medium. This study shows that LmrP is a true secondary transporter. In the absence of a proton motive force, LmrP facilitates downhill fluxes of ethidium in both directions. These fluxes are inhibited by other substrates of LmrP. The cysteine-reactive agent p-chloromercuri-benzene sulfonate inhibits these fluxes in wild type LmrP but not in the cysteine-less LmrP C270A mutant. Cysteine mutagenesis of LmrP resulted in three mutants, D68C/C270A, D128C/C270A, and E327C/C270A, with an energy-uncoupled phenotype. Asp68 is located in the conserved motif GXXX(D/E)(R/K)XGRK for the major facilitator superfamily of secondary transporters and was found to play an important role in energy coupling, whereas the negatively charged residues Asp128 and Glu327 have indirect effects on the transport process. L. lactis strains expressing these uncoupled mutants of LmrP show an increased rate of ethidium influx and an increased drug susceptibility compared with cells harboring an empty vector. The rate of influx in these mutants is enhanced by a transmembrane electrical potential, inside negative. These observations suggest a new strategy for eliminating drug-resistant microbial pathogens, i.e. the design and use of modulators of secondary multidrug resistance transporters that uncouple drug efflux from proton influx, thereby allowing transmembrane electrical potential driven influx of cationic drugs. PMID- 14561762 TI - A comparison of the self-association behavior of the plant cyclotides kalata B1 and kalata B2 via analytical ultracentrifugation. AB - The recently discovered cyclotides kalata B1 and kalata B2 are miniproteins containing a head-to-tail cyclized backbone and a cystine knot motif, in which disulfide bonds and the connecting backbone segments form a ring that is penetrated by the third disulfide bond. This arrangement renders the cyclotides extremely stable against thermal and enzymatic decay, making them a possible template onto which functionalities can be grafted. We have compared the hydrodynamic properties of two prototypic cyclotides, kalata B1 and kalata B2, using analytical ultracentrifugation techniques. Direct evidence for oligomerization of kalata B2 was shown by sedimentation velocity experiments in which a method for determining size distribution of polydisperse molecules in solution was employed. The shape of the oligomers appears to be spherical. Both sedimentation velocity and equilibrium experiments indicate that in phosphate buffer kalata B1 exists mainly as a monomer, even at millimolar concentrations. In contrast, at 1.6 mm, kalata B2 exists as an equilibrium mixture of monomer (30%), tetramer (42%), octamer (25%), and possibly a small proportion of higher oligomers. The results from the sedimentation equilibrium experiments show that this self-association is concentration dependent and reversible. We link our findings to the three-dimensional structures of both cyclotides, and propose two putative interaction interfaces on opposite sides of the kalata B2 molecule, one involving a hydrophobic interaction with the Phe6, and the second involving a charge-charge interaction with the Asp25 residue. An understanding of the factors affecting solution aggregation is of vital importance for future pharmaceutical application of these molecules. PMID- 14561763 TI - Oxidative phosphorylation and rotenone-insensitive malate- and NADH-quinone oxidoreductases in Plasmodium yoelii yoelii mitochondria in situ. AB - Respiration, membrane potential, and oxidative phosphorylation of mitochondria of Plasmodium yoelii yoelii trophozoites were assayed in situ after permeabilization with digitonin. ADP induced an oligomycin-sensitive transition from resting to phosphorylating respiration in the presence of oxidizable substrates. A functional respiratory chain was demonstrated. In addition, the ability of the parasite to oxidize exogenous NADH, as well as the insensitivity of respiration to rotenone and its sensitivity to flavone, suggested the presence of an alternative NADH-quinone (NADH-Q) oxidoreductase. Rotenone-insensitive respiration and membrane potential generation in the presence of malate suggested the presence of a malate-quinone oxidoreductase. These results are in agreement with the presence of genes in P. yoelii encoding for proteins with homology to NADH-Q oxidoreductases of bacteria, plant, fungi, and protozoa and malate-quinone oxidoreductases of bacteria. The complete inhibition of respiration by antimycin A and cyanide excluded the presence of an alternative oxidase as described in other parasites. An uncoupling effect of fatty acids was partly reversed by bovine serum albumin and GTP but was unaffected by carboxyatractyloside. These results provide the first biochemical evidence of the presence of an alternative NADH-Q oxidoreductase and a malate-quinone oxidoreductase and confirm the operation of oxidative phosphorylation in malaria parasites. PMID- 14561764 TI - The oncogene Nup98-HOXA9 induces gene transcription in myeloid cells. AB - The nucleoporin Nup98 gene is frequently rearranged in acute myelogenous leukemia (AML). In most cases this results in fusion of the N terminus of Nup98 to the DNA binding domain of a homeodomain transcription factor. The prototype of these fusions, Nup98-HOXA9, is associated with human AML and induces AML in mouse models. To understand the mechanisms by which Nup98-HOXA9 causes AML, we expressed it in myeloid cells and identified its target genes using high density oligonucleotide microarrays. The analysis was performed in triplicate and was confirmed by quantitative real time PCR. Of the 102 Nup98-HOXA9 target genes identified, 92 were up-regulated, and only 10 were down-regulated, suggesting a transcriptional activation function. A similar analysis of wild-type HOXA9 revealed 13 target genes, 12 of which were up-regulated, and 1 was down regulated. In contrast, wild-type Nup98 had no effect on gene expression, demonstrating that the HOXA9 DNA binding domain is required for gene regulation. Co-transfection experiments using a luciferase reporter linked to the promoter of one of the Nup98-HOXA9 target genes confirmed up-regulation at the transcriptional level by Nup98-HOXA9 but not by either HOXA9 or Nup98. These data indicate that Nup98-HOXA9 is an aberrant transcription factor whose activity depends on the HOXA9 DNA binding domain but has a stronger and wider transcriptional effect than HOXA9. Several of the genes regulated by Nup98-HOXA9 are associated with increased cell proliferation and survival as well as drug metabolism, providing insights into the pathogenesis and epidemiology of Nup98 HOXA9-induced AML. PMID- 14561765 TI - Molecular genetic analysis of human herpes virus 8-encoded viral FLICE inhibitory protein-induced NF-kappaB activation. AB - The human herpes virus 8 (HHV8)-encoded viral FLICE inhibitory protein (vFLIP), also known as K13, is known to activate the NF-kappaB pathway, a property not shared by other vFLIPs. Previous studies have demonstrated that HHV8 vFLIP K13 interacts with several cellular signaling proteins involved in NF-kappaB activation, such as receptor-interacting protein, NF-kappaB-inducing kinase, IkappaB kinase (IKK) 1, IKK2, and NF-kappaB essential modulator (NEMO). In this report we have used cell lines deficient in the above proteins to investigate the mechanism of NF-kappaB activation via HHV8 vFLIP K13. We demonstrate that receptor-interacting protein and NF-kappaB-inducing kinase are dispensable for vFLIP K13-induced NF-kappaB DNA binding and transcriptional activation. On the other hand, vFLIP K13-induced NF-kappaB DNA binding activity is significantly reduced, although not absent, in cells deficient in IKK1, IKK2, and NEMO. Furthermore, vFLIP K13-induced NF-kappaB transcriptional activity is only weakly present in IKK1-deficient cells and almost completely absent in those deficient in IKK2 and NEMO. HHV8 vFLIP K13-induced NF-kappaB activation in IKK1- and IKK2 deficient fibroblasts could be rescued by wild type but not by the kinase inactive mutants of IKK1 and IKK2, respectively. Consistent with the above results, vFLIP K13-induced NF-kappaB activation could be effectively blocked by chemical inhibitors of the kinase activity of IKK1 and IKK2. Thus, a cooperative interaction of all three subunits of the IKK complex is required for maximal NF kappaB activation via HHV8 vFLIP K13. Selective inhibitors of the IKK1 kinase activity may have a role in the treatment of disorders caused by abnormal NF kappaB activation by HHV8 vFLIP K13. PMID- 14561766 TI - Implication of DNA polymerase lambda in alignment-based gap filling for nonhomologous DNA end joining in human nuclear extracts. AB - Accurate repair of free radical-mediated DNA double-strand breaks by the nonhomologous end joining pathway requires replacement of fragmented nucleotides in the aligned ends by a gap-filling DNA polymerase. Nuclear extracts of human HeLa cells, supplemented with recombinant XRCC4-DNA ligase IV complex (XRCC4/ligase IV), were capable of accurately rejoining model double-strand break substrates with a 1- or 2-base gap, and the gap-filling step was dependent on XRCC4/ligase IV. To determine what polymerase was responsible for gap filling, end joining was examined in the presence of polyclonal antibodies against each of two prime candidate enzymes, DNA polymerases mu and lambda, both of which were present in the extracts. For a DNA substrate with partially complementary 3' overhangs and a 2-base gap, antibodies to polymerase lambda completely eliminated both gap filling and accurate end joining, whereas antibodies to polymerase mu had little effect. Immunodepletion of polymerase lambda, but not polymerase mu, likewise blocked both gap filling and end joining, and both functions could be restored by addition of recombinant polymerase lambda. Recombinant polymerase mu, and a truncated polymerase lambda lacking the Brca1 C-terminal domain, were at least 10-fold less active in restoring gap filling to the immunodepleted extracts, and polymerase beta was completely inactive. The results suggest that polymerase lambda is the primary gap-filling polymerase for accurate nonhomologous end joining, and that the Brca1 C-terminal domain is required for this activity. PMID- 14561767 TI - HIV-1 Vpu sequesters beta-transducin repeat-containing protein (betaTrCP) in the cytoplasm and provokes the accumulation of beta-catenin and other SCFbetaTrCP substrates. AB - The human immunodeficiency virus type 1 Vpu protein acts as an adaptor for the proteasomal degradation of CD4 by recruiting CD4 and beta-transducin repeat containing protein (betaTrCP), the receptor component of the multisubunit SCF betaTrCP E3 ubiquitin ligase complex. We showed that the expression of a Vpu green fluorescent fusion protein prevented the proteosomal degradation of betaTrCP substrates such as beta-catenin, IkappaBalpha, and ATF4, which are normally directly targeted to the proteasome for degradation. Beta-catenin was translocated into the nucleus, whereas the tumor necrosis factor-induced nuclear translocation of NFkappaB was impaired. Beta-catenin was also up-regulated in cells producing Vpu+ human immunodeficiency virus type 1 but not in cells producing Vpu-deficient viruses. The overexpression of ATF4 also provoked accumulation of beta-catenin, but to a lower level than that resulting from the expression of Vpu. Finally, the expression of Vpu induces the exclusion of betaTrCP from the nucleus. These data suggest that Vpu is a strong competitive inhibitor of betaTrCP that impairs the degradation of SCFbetaTrCP substrates as long as Vpu has an intact phosphorylation motif and can bind to betaTrCP. PMID- 14561768 TI - The crystal structure of the Calystegia sepium agglutinin reveals a novel quaternary arrangement of lectin subunits with a beta-prism fold. AB - The high number of quaternary structures observed for lectins highlights the important role of these oligomeric assemblies during carbohydrate recognition events. Although a large diversity in the mode of association of lectin subunits is frequently observed, the oligomeric assemblies of plant lectins display small variations within a single family. The crystal structure of the mannose-binding jacalin-related lectin from Calystegia sepium (Calsepa) has been determined at 1.37-A resolution. Calsepa exhibits the same beta-prism fold as identified previously for other members of the family, but the shape and the hydrophobic character of its carbohydrate-binding site is unlike that of other members, consistent with surface plasmon resonance analysis showing a preference for methylated sugars. Calsepa reveals a novel dimeric assembly markedly dissimilar to those described earlier for Heltuba and jacalin but mimics the canonical 12 stranded beta-sandwich dimer found in legume lectins. The present structure exemplifies the adaptability of the beta-prism building block in the evolution of plant lectins and highlights the biological role of these quaternary structures for carbohydrate recognition. PMID- 14561769 TI - Cardiolipin stabilizes respiratory chain supercomplexes. AB - Cardiolipin stabilized supercomplexes of Saccharomyces cerevisiae respiratory chain complexes III and IV (ubiquinol:cytochrome c oxidoreductase and cytochrome c oxidase, respectively), but was not essential for their formation in the inner mitochondrial membrane because they were found also in a cardiolipin-deficient strain. Reconstitution with cardiolipin largely restored wild-type stability. The putative interface of complexes III and IV comprises transmembrane helices of cytochromes b and c1 and tightly bound cardiolipin. Subunits Rip1p, Qcr6p, Qcr9p, Qcr10p, Cox8p, Cox12p, and Cox13p and cytochrome c were not essential for the assembly of supercomplexes; and in the absence of Qcr6p, the formation of supercomplexes was even promoted. An additional marked effect of cardiolipin concerns cytochrome c oxidase. We show that a cardiolipin-deficient strain harbored almost inactive resting cytochrome c oxidase in the membrane. Transition to the fully active pulsed state occurred on a minute time scale. PMID- 14561770 TI - Characterization of the agent of "high plains disease": mass spectrometry determines the sequence of the disease-specific protein. AB - The "32-kDa" protein specifically associated with high plains disease was characterized by time-of-flight mass spectrometry, after the agent had been isolated in pure culture by "vascular puncture inoculation," a novel mechanical means of transmission. Two isolates from different geographic locations each consisted of a mixture of subpopulations that were highly homologous to an amino acid sequence derived from a nucleotide sequence (U60141) deposited in GenBank trade mark by the Nebraska group as "the probable N-protein of high plains virus." However, the U60141 sequence was found to be incomplete; de novo sequencing of peptides produced by proteolytic digestions of the 32-kDa band from an SDS-PAGE separation showed that an additional 18 amino acid residues were present at the N terminus. BLAST (basic local alignment search tool) examination of the sequence showed no significant homology with any protein in the databases, indicating that the infectious agent of high plains disease is likely a member of a hitherto unclassified virus group. PMID- 14561771 TI - Ways of dying: multiple pathways to apoptosis. PMID- 14561772 TI - The species-specific egg receptor for sea urchin sperm adhesion is EBR1,a novel ADAMTS protein. AB - Species-specific adhesion of sperm to the egg during sea urchin fertilization involves the interaction of the sperm adhesive protein,bindin, and a complementary receptor on the egg surface,and serves to restrict the gene pool to individuals of the same species. We used PCR representation difference analysis to clone the species-specific egg receptor for bindin, EBR1, from Strongylocentrotus franciscanus (Sf) and S. purpuratus (Sp). Sf-EBR1 contains a novel ADAMTS-like N-terminal domain followed by approximately 19 tandem EBR repeats consisting of alternating CUB and thrombospondin type 1 (TSP-1) domains where the last 10 EBR repeats are species-specific and highly conserved. Recombinant protein corresponding to the species-specific EBR repeat displays species-specific sperm adhesion and bindin-binding activity. The Sp-EBR1 ortholog has the same ADAMTS (a disintegrin and metalloprotease with thrombospondin type-1 modules) core region followed by eight and one-half tandem egg bindin receptor (EBR) repeats that share 88% identity with the Sf-EBR1 repeats,but has an entirely different species-specific domain consisting of hyalin-like (HYR) repeats. Thus,the species-specific domains of egg bindin receptor 1 (EBR1) from both species function as the egg surface receptor to mediate species-specific sperm adhesion. PMID- 14561773 TI - Model of the brain tumor-Pumilio translation repressor complex. AB - The Brain Tumor (Brat) protein is recruited to the 3' untranslated region (UTR) of hunchback mRNA to regulate its translation. Recruitment is mediated by interactions between the Pumilio RNA-binding Puf repeats and the NHL domain of Brat, a conserved structural motif present in a large family of growth regulators. In this report, we describe the crystal structure of the Brat NHL domain and present a model of the Pumilio-Brat complex derived from in silico docking experiments and supported by mutational analysis of the protein-protein interface. A key feature of the model is recognition of the outer, convex surface of the Pumilio Puf domain by the top, electropositive face of the six-bladed Brat beta-propeller. In particular, an extended loop in Puf repeat 8 fits in the entrance to the central channel of the Brat beta-propeller. Together, these interactions are likely to be prototypic of the recruitment strategies of other NHL-containing proteins in development. PMID- 14561774 TI - The Drosophila Ste20 family kinase dMST functions as a tumor suppressor by restricting cell proliferation and promoting apoptosis. AB - In a genetic screen for mutations that restrict cell growth and organ size, we identified a new tumor suppressor gene, dMST, which encodes the Drosophila homolog of the mammalian Ste20 kinase family members MST1 and MST2. Loss-of function mutations in dMST result in overgrown tissues containing more cells of normal size. dMST mutant cells exhibit elevated levels of Cyclin E and DIAP1, increased cell growth and proliferation, and impaired apoptosis. dMST forms a complex with Sav and Wts, two tumor suppressors also implicated in regulating both cell proliferation and apoptosis, suggesting that they act in common pathways. PMID- 14561775 TI - The spindle checkpoint requires cyclin-dependent kinase activity. AB - The spindle checkpoint prevents anaphase onset until completion of mitotic spindle assembly by restraining activation of the ubiquitin ligase anaphase promoting complex/cyclosome-Cdc20 (APC/CCdc20). We show that the spindle checkpoint requires mitotic cyclin-dependent kinase (cdk) activity. Inhibiting cdk activity overrides checkpoint-dependent arrest in Xenopus egg extracts and human cells. Following inhibition, the interaction between APC/C and Cdc20 transiently increases while the inhibitory checkpoint protein Mad2 dissociates from Cdc20. Cdk inhibition also overcomes Mad2-induced mitotic arrest. In addition, in vitro cdk1-phosphorylated Cdc20 interacts with Mad2 rather than APC/ C. Thus, cdk activity is required to restrain APC/CCdc20 activation until completion of spindle assembly. PMID- 14561776 TI - Regulation of the transcriptional activator NtrC1: structural studies of the regulatory and AAA+ ATPase domains. AB - Transcription by sigma54 RNA polymerase depends on activators that contain ATPase domains of the AAA+ class. These activators, which are often response regulators of two-component signal transduction systems, remodel the polymerase so that it can form open complexes at promoters. Here, we report the first crystal structures of the ATPase domain of an activator, the NtrC1 protein from the extreme thermophile Aquifex aeolicus. This domain alone, which is active, crystallized as a ring-shaped heptamer. The protein carrying both the ATPase and adjacent receiver domains, which is inactive, crystallized as a dimer. In the inactive dimer, one residue needed for catalysis is far from the active site, and extensive contacts among the domains prevent oligomerization of the ATPase domain. Oligomerization, which completes the active site, depends on surfaces that are buried in the dimer, and hence, on a rearrangement of the receiver domains upon phosphorylation. A motif in the ATPase domain known to be critical for coupling energy to remodeling of polymerase forms a novel loop that projects from the middle of an alpha helix. The extended, structured loops from the subunits of the heptamer localize to a pore in the center of the ring and form a surface that could contact sigma54. PMID- 14561777 TI - Tousled-like kinase functions with the chromatin assembly pathway regulating nuclear divisions. AB - Tousled-like kinases (TLKs) constitute a family of serine/threonine kinases conserved in plants and animals that act in a cell cycle-dependent manner. In mammals, their activity peaks during S phase, when they phosphorylate the antisilencing function protein 1 (ASF1), a histone chaperone involved in replication-dependent chromatin assembly. Here, we show that Drosophila ASF1 is also a phosphorylation target of TLK, and that the two components cooperate to control chromatin replication in vivo. By altering TLK activity through loss-of function mutations, we show that nuclear divisions are arrested at interphase, followed by apoptosis. Overexpression of TLK alters the chromatin structure, suggesting that TLK mediates the activity of chromatin proteins. These results suggest that TLK coordinates cell cycle progression through the regulation of chromatin dynamics. PMID- 14561779 TI - PAX6 and congenital eye malformations. AB - The PAX6 gene is a paradigm for our understanding of the molecular genetics of mammalian eye development. Twelve years after its identification it is one of the most intensively studied genes, both in terms of its diverse and complex functions during oculogenesis and its role in an ever-increasing variety of human congenital eye malformations. The PAX6 field has benefited greatly from the continued input of clinicians, human geneticists and developmental biologists. This review summarizes the latest data on the PAX6 mutation spectrum and recent insights into Pax6 function from the mouse. PMID- 14561778 TI - Opposing actions of Arx and Pax4 in endocrine pancreas development. AB - Genes encoding homeodomain-containing proteins potentially involved in endocrine pancreas development were isolated by combined in silico and nested-PCR approaches. One such transcription factor, Arx, exhibits Ngn3-dependent expression throughout endocrine pancreas development in alpha, beta-precursor, and delta cells. We have used gene targeting in mouse embryonic stem cells to generate Arx loss-of-function mice. Arx-deficient animals are born at the expected Mendelian frequency, but develop early-onset hypoglycemia, dehydration, and weakness, and die 2 d after birth. Immunohistological analysis of pancreas from Arx mutants reveals an early-onset loss of mature endocrine alpha cells with a concomitant increase in beta-and delta-cell numbers, whereas islet morphology remains intact. Our study indicates a requirement of Arx for alpha-cell fate acquisition and a repressive action on beta-and delta-cell destiny, which is exactly the opposite of the action of Pax4 in endocrine commitment. Using multiplex reverse transcriptase PCR (RT-PCR), we demonstrate an accumulation of Pax4 and Arx transcripts in Arx and Pax4 mutant mice, respectively. We propose that the antagonistic functions of Arx and Pax4 for proper islet cell specification are related to the pancreatic levels of the respective transcripts. PMID- 14561780 TI - The effect of massive small bowel resection and oral epidermal growth factor therapy on SGLT-1 distribution in rabbit distal remnant. AB - Small bowel resection decreases brush border membrane (BBM) glucose uptake kinetics. Oral epidermal growth factor (EGF) returns net glucose transport across intact tissue to control levels despite persistence of a defect in BBM glucose uptake. The purpose of this study was to examine the effects of resection and EGF treatment on sodium-dependent glucose cotransporter 1 (SGLT-1) expression in distal remnant tissue. New Zealand White rabbits (1 kg) underwent 70% small bowel resection (R). One group of resected animals (R-EGF) received oral EGF (40 microg/kg, days 3-8). Distal remnant tissue was harvested 10 d after surgery, and compared with controls (C). Mucosal SGLT-1 mRNA was measured by Northern blot, BBM SGLT-1 content by Western blot, and villus distribution of SGLT-1 protein and mRNA by immunofluorescence and in situ hybridization. Western blot indicated BBM from both resected and EGF-treated tissue had decreased SGLT-1 content (C, 0.55 +/- 0.04; R, 0.35 +/- 0.04; R-EGF, 0.35 +/- 0.03 trace OD; n = 5; p < 0.05). Northern blot revealed no alterations in mucosal SGLT-1 mRNA content in any group. SGLT-1 protein and mRNA localization in control tissues was characterized by a gradual increase in stain intensity from the base of the villus to the villus tip. Resection altered SGLT-1 protein and mRNA expression along the villus axis with intensity being strongest in the mid-villus region and little expression at the tip of the villus. Oral EGF normalized SGLT-1 protein and mRNA expression to control patterns. Resection alters SGLT-1 protein and mRNA expression along the villus axis, despite no change in total mucosal SGLT-1 mRNA content. EGF normalized villus SGLT-1 protein and mRNA distribution, without altering overall BBM SGLT-1 content or mucosal mRNA levels. PMID- 14561781 TI - Therapeutic hypercapnia is not protective in the in vivo surfactant-depleted rabbit lung. AB - Permissive hypercapnia because of reduced tidal volume is associated with improved survival in lung injury, whereas therapeutic hypercapnia-deliberate elevation of arterial Pco2-protects against in vivo reperfusion injury and injury produced by severe lung stretch. No published studies to date have examined the effects of CO2 on in vivo models of neonatal lung injury. We used an established in vivo rabbit model of surfactant depletion to investigate whether therapeutic hypercapnia would improve oxygenation and protect against ventilator-induced lung injury. Animals were randomized to injurious (tidal volume, 12 mL/kg; positive end-expiratory pressure, 0 cm H2O) or protective ventilatory strategy (tidal volume, 5 mL/kg; positive end-expiratory pressure, 12.5 cm H2O), and to receive either control conditions or therapeutic hypercapnia (fraction of inspired CO2, 0.12). Oxygenation (alveolar-arterial O2 difference, arterial Po2), lung injury (alveolar-capillary protein leak, impairment of static compliance), and selected bronchoalveolar lavage and plasma cytokines (IL-8, growth-related oncogene, monocyte chemoattractant protein-1, and tumor necrosis factor-alpha) were measured. Injurious ventilation resulted in a large alveolar-arterial O2 gradient, elevated peak airway pressure, increased protein leak, and impaired lung compliance. Therapeutic hypercapnia did not affect any of these outcomes. Tumor necrosis factor-alpha was not increased by mechanical stretch in any of the groups. Therapeutic hypercapnia abolished the stretch-induced increase in bronchoalveolar lavage monocyte chemoattractant protein-1, but did not affect any of the other mediators studied. Therapeutic hypercapnia may attenuate the impairment in oxygenation and inhibit certain cytokines. Because hypercapnia inhibits certain cytokines but does not alter lung injury, the pathogenic role of these cytokines in lung injury is questionable. PMID- 14561782 TI - The role of PKCdelta and PKCepsilon in the neonatal rat colon in response to hypoxia challenge. AB - Previous studies have determined that, in response to bacterial endotoxin, the colonic mucosa of the 10-d-old neonatal rat was more susceptible to injury than was the colon of the 25-d-old mature animal. Furthermore, it is known that certain isoforms of protein kinase C (PKC), specifically PKCdelta and PKCepsilon, mediate intestinal inflammatory responses to specific challenges. Therefore, in the present study, we have examined the association between the activation of these PKC isoforms and the enhanced susceptibility to hypoxia-induced challenge. In response to exposure to a hypoxic environment (14% O2/86% N2, 30 min), the degree of inflammation and tissue damage was significantly greater in 10- than in 25-d-old rats. The injury in 10-d-old rats was associated with activation of PKCdelta and PKCepsilon as estimated by translocation of the isoform from cytosolic to membrane fraction of the tissue lysate. There was no activation of either isoform in colons from 25-d-old rats after hypoxia. Pretreatment of 10-d old rats with epidermal growth factor (EGF) (10 microg/kg) but not 16,16 dimethyl prostaglandin E2 (2 microg/kg) significantly reduced the extent of colonic injury, whereas neither agent was able to exert significant protection of the colonic mucosa of 25-d-old rats. PKC activation associated with hypoxia was not evident after EGF treatment in 10-d-old rats. In 25-d-old rats, prostaglandin E2 treatment was linked with an activation of PKCepsilon only. In conclusion, these data suggest that activation of PKCdelta and PKCepsilon is associated with the enhanced susceptibility to injury evident in suckling neonatal rat colon. EGF mediated protection of the colon in these animals results in a removal of this PKC activation. PMID- 14561783 TI - The effects of systemic magnesium sulfate infusion on brain magnesium concentrations and energy state during hypoxia-ischemia in newborn miniswine. AB - The mechanism of neuroprotection associated with systemically administered magnesium remains unclear. This investigation examined the acute effects of systemically administered MgSO4 on brain extracellular ([Mg]ecf) and intracellular ([Mg]i) fluid Mg concentrations, specific brain phosphorylated metabolites, and brain intracellular pH. Miniswine were studied with P-31 magnetic resonance spectra, to derive [Mg]i, and brain microdialysis probes, to measure [Mg]ecf. Animals were infused with MgSO4 (n = 5, 275 mg/kg over 30 min followed by 100 mg/kg over 30 min, designated MgHI) or Na2SO4 (n = 5, designated NaHI), and both groups underwent hypoxia-ischemia (HI) over the last 15 min of the infusions. Groups differed in plasma [Mg] at the completion of HI (9.1 +/- 1.5 versus 1.1 +/- 0.6 mM for MgHI and NaHI, respectively, p < 0.05). MgHI had elevations of [Mg]ecf (0.23 +/- 0.11 and 0.40 +/- 0.14 mM at control and completion of HI, respectively), and [Mg]ecf was unchanged for NaHI (p < 0.05 versus MgHI). At the completion of HI, MgHI had greater decreases in nucleoside triphosphate (NTP) (48 +/- 6% of control), and more brain acidosis after HI (6.01 +/- 0.07) compared with NaHI (NTP, 70 +/- 3% of control; brain pH, 6.51 +/- 0.14, both p < 0.05 versus MgHI). [Mg]i increased to elevated values during HI in both MgHI and NaHI (p < 0.05 versus control of each group) and remained higher in MgHI over the next 25 min (p < 0.05 versus NaHI). There were inverse correlations during HI between [Mg]i and brain NTP (r2 = 0.73 and 0.59 for MgHI and NaHI, respectively), and brain acidosis (r2 = 0.85 and 0.85 for MgHI and NaHI, respectively) in each group. These findings indicate complex effects of Mg on the brain. Elevation of [Mg]ecf may be beneficial with regards to excitatory neurotransmitters. However, greater disturbance of brain NTP concentration, more acidosis, and the increase in [Mg]i may offset any benefit. The results warrant further investigation using indicators of neuronal injury to determine whether Mg supplementation provides neuroprotection. PMID- 14561784 TI - Hypoxic oxygen fluctuations produce less severe retinopathy than hyperoxic fluctuations in a rat model of retinopathy of prematurity. AB - The aim of this study was to investigate whether the mean around which arterial oxygen fluctuations take place was important in a unique animal model of oxygen induced retinopathy. Retinopathy of prematurity (ROP) is associated with fluctuating arterial oxygen. A recent retrospective study suggested that management of high-risk preterm infants at lower oxygen saturations was associated with less severe ROP. Rat pups were raised in a variable oxygen environment around a high (24%), normal (21%) or low (17%) mean inspired oxygen for 14 d. Rat pups raised in the high (24%) mean variable oxygen environment had more retarded retinal vascular development than did rats raised in an environment that fluctuated around 21% mean oxygen. In contrast, rats raised in a lower mean (17%) but still variable oxygen environment had no discernible retinal differences from controls raised in constant room air. Rats raised in a relatively hypoxic but variable oxygen environment develop less severe retinal vascular abnormalities than those raised in variable oxygen around higher oxygen means. PMID- 14561785 TI - Bone marrow stroma damage induced by chemotherapy for acute lymphoblastic leukemia in children. AB - Several studies have suggested a role of bone marrow stroma injury in long-term chemotherapy-induced hematopoietic failure. To evaluate whether bone marrow microenvironment is altered by chemotherapy for acute lymphoblastic leukemia (ALL) and to determine its contribution to postchemotherapy anemia, we investigated the ability of stroma from children receiving maintenance chemotherapy for ALL to support hematopoiesis. Long-term bone marrow cultures (LTBMC) were established with bone marrow cells either from ALL children under therapy (n = 24) or from control subjects (n = 19). Nonadherent cells and colony forming units-granulocytic monocytic (CFU-GM) output in LTBMC did not differ between patients and controls. In contrast, burst forming unit-erythroid (BFU-E) numbers were lower in patient LTBMC (p = 0.013). Co-cultures of normal CD34+ cells and preformed patient or control stromas showed significantly reduced hematopoietic supportive capabilities of patient stromas: both CFU-GM and BFU-E were reduced (p = 0.002 and 0.046, respectively). In addition, supernatants (SN) of patients' LTBMC inhibited normal BFU-E growth compared with SN of normal LTBMC. Transforming growth factor (TGF)-beta1 levels were increased in patient cultures (p = 0.0039) and inversely correlated with BFU-E produced in LTBMC (r = 0.36, p = 0.04). Neutralization of TGF-beta1 significantly increased the BFU-E output of patient LTBMC (p = 0.0078). In contrast, macrophage inflammatory peptide (MIP)-1alpha levels were lower in SN of patients compared with controls (p = 0.015). Thus, chemotherapy for ALL induces functional deregulation within bone marrow stromal cells with an increase in the growth-inhibiting factor TGF beta1, together with a decrease in MIP-1alpha, which might contribute to hematopoietic toxicity. PMID- 14561786 TI - Increased brain levels of F2-isoprostane are an early marker of behavioral sequels in a rat model of global perinatal asphyxia. AB - Perinatal asphyxia is a major cause of immediate and postponed brain damage in the newborn. It may be responsible for several delayed neurologic disorders and, in this respect, early markers of brain injury would be relevant for therapeutic intervention as well as for identification of infants at high risk for developmental disabilities. Biochemical measurements (brain F2-isoprostane levels) and behavioral tests (ultrasonic vocalization pattern on postnatal days (pnd) 5, 8, and 11, spontaneous motor behaviors on pnd 7 and 12, and homing response on pnd 10) were performed in a rat model of global perinatal asphyxia in the immature neonate. Caesarean section was performed in rats and the pups, still in uterus horns, were placed into a water bath at 37 degrees C for either 10 or 20 min. Caesarean delivered pups were used as controls. Pups experiencing severe (20 min), in contrast to those undergoing the 10 min, asphyctic insult presented with detectable abnormalities including early (two hours after the insult) increase in brain F2-isoprostane (a direct marker of oxidative injury) without detectable changes in PGE2, COX-2 and iNOS levels, and delayed physical (reduced weight gain on pnd 5 and thereafter) and behavioral disturbances (alterations in ultrasound emission on pnd 11 and spontaneous motricity levels mainly). These findings suggest that increased brain F2-isoprostane levels shortly after the asphyctic insult are predictive of delayed behavioral disturbances in the newborn rat. The present 20-min asphyxia model might serve for the assessment of preventive and curative strategies to treat neurologic/behavioral disturbances associated with perinatal asphyxia. PMID- 14561787 TI - A new method for the measurement of cerebral blood volume and total circulating blood volume using near infrared spatially resolved spectroscopy and indocyanine green: application and validation in neonates. AB - A new technique known as tissue dye densitometry (TDD) has been developed to simultaneously measure cerebral blood volume (CBV) and total circulating blood volume (TCV) using near infrared (NIR) spatially resolved spectroscopy (SRS) and the injection of indocyanine green (ICG). Using a medical NIR spectrometer with SRS capability (NIRO-300, Hamamatsu KK), a new parameter is calculated known as the ICG Hb index (IHI), which represents the ratio of ICG concentration to Hb concentration in tissue. Acting as a tracer, ICG is cleared by the liver over 15 min, providing a change of tracer concentration (DeltaCICG,tis), which allows the calculation of the total Hb concentration in tissue (tcHb) using the equation: tcHbtis (micro molar) = DeltaCICG,tis/DeltaIHI. The CBV can subsequently be calculated from tcHbtis given the absolute Hb concentration in blood (g/dL), from which the ICG concentration in blood (DeltaCICG,bl) is obtained. By back extrapolating the DeltaCICG,bl curve to the peak time, the initial ICG concentration in tissue blood (C0ICG,bl) can be found and TCV can then be calculated. The TCV of 17 neonates were measured using the TDD technique and for comparison using the previously reported fetal Hb dilution technique (FHD). The mean TCV measured by the FHD and TDD techniques were 70.19 +/- 13.73 mL/kg and 70.80 +/- 32.54 mL/kg. The Bland Altman plot showed that the bias was 0.61 +/- 34.34 mL/kg and limits of agreement (2 SD) were -68.07 mL/kg and 69.30 mL/kg. The agreement is limited and the TDD technique needs further validation and development for use in a clinical environment. PMID- 14561788 TI - Immunoglobulins in otitis-prone children. AB - Defective or immature antibody responses to pathogens in children may explain the increased susceptibility to acute otitis media (AOM) in otitis-prone children. In literature, data on immunology have been based on studies of small groups of severely otitis-prone children and have not been consistent. Humoral immune status was assessed in 365 children, 1-7 years old, with two or more documented episodes of AOM in the previous year. Children with 4 or more episodes in the preceding year were defined as otitis-prone. Serum immunoglobulin levels were determined by radial immunodiffusion. Immunoglobulin levels of otitis-prone children were compared with those of children who had experienced 2-3 AOM episodes per year. Children with recurrent episodes of AOM were found to have normal or increased serum IgA, IgM, IgG, and IgG1 levels compared with normal values for age, whereas the serum IgG2 levels were mostly in the lower normal range. Twenty-two percent of all children showed IgG2 levels lower than 2 SD below the age-specific mean. Interestingly, the otitis-prone group of children showed significantly lower median and mean levels for all immunoglobulins compared with those children with only 2-3 previous AOM episodes. Lower immunoglobulin levels in otitis-prone children suggest a generalized decreased antibody response in otitis-prone children. PMID- 14561789 TI - How often should we screen for cervical cancer? PMID- 14561790 TI - Two worlds of malaria. PMID- 14561791 TI - Autoantibodies in systemic lupus erythematosus--there before you know it. PMID- 14561792 TI - Risk of cervical cancer associated with extending the interval between cervical cancer screenings. AB - BACKGROUND: Although contemporary guidelines suggest that the intervals between Papanicolaou tests can be extended to three years among low-risk women with previous negative tests, the excess risk of cervical cancer associated with less frequent than annual screening is uncertain. METHODS: We determined the prevalence of biopsy-proven cervical neoplasia among 938,576 women younger than 65 years of age, stratified according to the number of previous consecutive negative Papanicolaou tests. Using a Markov model that estimates the rate at which dysplasia will progress to cancer, we estimated the risk of cancer within three years after one or more negative Papanicolaou tests, as well as the number of additional Papanicolaou tests and colposcopic examinations that would be required to avert one case of cancer given a particular interval between screenings. RESULTS: Among 31,728 women 30 to 64 years of age who had had three or more consecutive negative tests, the prevalence of biopsy-proven cervical intraepithelial neoplasia of grade 2 was 0.028 percent and the prevalence of grade 3 neoplasia was 0.019 percent; none of the women had invasive cervical cancer. According to our model, the estimated risk of cancer with annual Papanicolaou tests for three years was 2 in 100,000 among women 30 to 44 years of age, 1 in 100,000 among women 45 to 59 years of age, and 1 in 100,000 among women 60 to 64 years of age; these risks would be 5 in 100,000, 2 in 100,000, and 1 in 100,000, respectively, if screening were performed once three years after the last negative test. To avert one additional case of cancer by screening 100,000 women annually for three years rather than once three years after the last negative test, an average of 69,665 additional Papanicolaou tests and 3861 colposcopic examinations would be needed in women 30 to 44 years of age and an average of 209,324 additional Papanicolaou tests and 11,502 colposcopic examinations in women 45 to 59 years of age. CONCLUSIONS: As compared with annual screening for three years, screening performed once three years after the last negative test in women 30 to 64 years of age who have had three or more consecutive negative Papanicolaou tests is associated with an average excess risk of cervical cancer of approximately 3 in 100,000. PMID- 14561793 TI - Delayed onset of malaria--implications for chemoprophylaxis in travelers. AB - BACKGROUND: Most antimalarial agents used by travelers act on the parasite's blood stage and therefore do not prevent late-onset illness, particularly that due to species that cause relapsing malaria. We examined the magnitude of this problem among Israeli and American travelers. METHODS: We examined malaria surveillance data from Israel and the United States to determine the traveler's destination, the infecting species, the type of chemoprophylaxis used, and the incubation period. RESULTS: In Israel, from 1994 through 1999, there were 300 cases of malaria among returning travelers in which one species of plasmodium could be identified. In 134 of these cases (44.7 percent), the illness developed more than two months after the traveler's return; nearly all of these cases were due to infection with Plasmodium vivax or P. ovale. In 108 of the 134 cases (80.6 percent), the patient had used an antimalarial regimen according to national guidelines. In the United States, from 1992 through 1998, there were 2822 cases of malaria among travelers in which the cause could be evaluated. Late illness developed in 987 (35.0 percent) of these travelers. The infection was due to P. vivax in 811 travelers, P. ovale in 66, P. falciparum in 59, and P. malariae in 51; 614 (62.2 percent) of those with late-onset illness had appropriately taken an effective antimalarial agent. CONCLUSIONS: In more than one third of malaria infected travelers, the illness developed more than two months after their return. Most of these late-onset illnesses are not prevented by the commonly used and effective blood schizonticides. Agents that act on the liver phase of malaria parasites are needed for more effective prevention of malaria in travelers. PMID- 14561794 TI - Early malignant progression of hereditary medullary thyroid cancer. AB - BACKGROUND: An age-related progression from C-cell hyperplasia to medullary thyroid carcinoma is associated with various germ-line mutations in the rearranged during transfection (RET) proto-oncogene that could be used to identify the optimal time for prophylactic surgery. METHODS: In this European multicenter study conducted from July 1993 to February 2001, we enrolled patients who had a RET point mutation in the germ line, were 20 years of age or younger, were asymptomatic, and had undergone total thyroidectomy after confirmation of the RET mutation. Exclusion criteria were medullary thyroid carcinomas of more than 10 mm in greatest dimension and distant metastasis. RESULTS: Altogether, 207 patients from 145 families were identified. There was a significant age-related progression from C-cell hyperplasia to medullary thyroid carcinoma and, ultimately, nodal metastasis in patients whose RET mutations were grouped according to the extracellular- and intracellular-domain codons affected and in those with the codon 634 genotype. No lymph-node metastases were noted in patients younger than 14 years of age. The age-related penetrance was unaffected by the type of amino acid substitution encoded by the various codon 634 mutations. The codon-specific differences in the age at presentation of cancer and the familial rates of concomitant adrenal and parathyroid involvement suggest that the risk of progression was based on the transforming potential of the individual RET mutation. CONCLUSIONS: These data provide initial guidelines for the timing of prophylactic thyroidectomy in asymptomatic carriers of RET gene mutations. PMID- 14561795 TI - Development of autoantibodies before the clinical onset of systemic lupus erythematosus. AB - BACKGROUND: Although much is known about the natural history of systemic lupus erythematosus (SLE), the development of SLE autoantibodies before the diagnosis of the disease has not been extensively explored. We investigated the onset and progression of autoantibody development before the clinical diagnosis. METHODS: The Department of Defense Serum Repository contains approximately 30 million specimens prospectively collected from more than 5 million U.S. Armed Forces personnel. We evaluated serum samples obtained from 130 persons before they received a diagnosis of SLE, along with samples from matched controls. RESULTS: In 115 of the 130 patients with SLE (88 percent), at least one SLE autoantibody tested was present before the diagnosis (up to 9.4 years earlier; mean, 3.3 years). Antinuclear antibodies were present in 78 percent (at a dilution of 1:120 or more), anti-double-stranded DNA antibodies in 55 percent, anti-Ro antibodies in 47 percent, anti-La antibodies in 34 percent, anti-Sm antibodies in 32 percent, anti-nuclear ribonucleoprotein antibodies in 26 percent, and antiphospholipid antibodies in 18 percent. Antinuclear, antiphospholipid antibodies, anti-Ro, and anti-La antibodies were present earlier than anti-Sm and anti-nuclear ribonucleoprotein antibodies (a mean of 3.4 years before the diagnosis vs. 1.2 years, P=0.005). Anti-double-stranded DNA antibodies, with a mean onset 2.2 years before the diagnosis, were found later than antinuclear antibodies (P=0.06) and earlier than anti-nuclear ribonucleoprotein antibodies (P=0.005). For many patients, the earliest available serum sample was positive; therefore, these measures of the average time from the first positive antibody test to the diagnosis are underestimates of the time from the development of antibodies to the diagnosis. Of the 130 initial matched controls, 3.8 percent were positive for one or more autoantibodies. CONCLUSIONS: Autoantibodies are typically present many years before the diagnosis of SLE. Furthermore, the appearance of autoantibodies in patients with SLE tends to follow a predictable course, with a progressive accumulation of specific autoantibodies before the onset of SLE, while patients are still asymptomatic. PMID- 14561796 TI - Images in clinical medicine. Intraabdominal pregnancy after hysterectomy. PMID- 14561797 TI - Primary progressive aphasia--a language-based dementia. PMID- 14561798 TI - Paraneoplastic syndromes involving the nervous system. PMID- 14561799 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 32-2003. A 37-year-old woman with atypical squamous cells on a Papanicolaou smear. PMID- 14561800 TI - Lessons learned from the management of a rare genetic cancer. PMID- 14561801 TI - Prevention of prostate cancer with finasteride. PMID- 14561802 TI - Renal arterial resistance index. PMID- 14561803 TI - Case 25-2003: congenital cytomegalovirus infection. PMID- 14561804 TI - The solitary pulmonary nodule. PMID- 14561805 TI - Aristolochic acid, an herbal carcinogen, sold on the Web after FDA alert. PMID- 14561806 TI - Images in clinical medicine. "T-U-P" syndrome, or pseudoatrial flutter. PMID- 14561807 TI - Antioxidant enzymatic defences in human follicular fluid: characterization and age-dependent changes. AB - The aim of this work was to study the antioxidant enzymatic defences in human follicular fluid and investigate their possible changes during reproductive ageing. To this end, we tested the specific activities and protein expression of enzymes involved in reactive oxygen species (ROS) scavenging and in detoxification of ROS byproducts in follicular fluid from young (range 27-32 years, n = 12) and older (range 39-45 years, n = 12) women participating in an IVF programme. Results show that all the tested enzymes [superoxide dismutase (SOD), catalase, glutathione peroxidase, glutathione transferase, glutathione reductase] were significantly expressed in human follicular fluid. However, when the two age groups were compared, we found that follicular fluid from older women exhibited a reduced level of glutathione transferase and catalase activities and a higher level of SOD activity. Immunoblot analysis revealed that ageing was associated with decreased protein expression of GST Pi isoform and did not affect SOD and catalase protein expression. Taken together, these findings indicate that reproductive ageing is accompanied by a change in the antioxidant enzymatic pattern that could impair ROS scavenging efficiency in the follicular environment. PMID- 14561808 TI - Multiple forms of redox activity in populations of human spermatozoa. AB - In this study we have examined the biochemical attributes of the redox systems that regulate human sperm function using 2-(4-iodophenyl)-3-(4-nitrophenyl)-5 (2,4-disulphophenyl)-2H-tetrazolium, monosodium salt (WST-1), lucigenin and luminol-peroxidase as probes. WST-1 was readily reduced by human spermatozoa in the presence of an intermediate electron acceptor (IEA) or NAD(P)H. The IEA mediated activity resembled a previously described trans-membrane NADH oxidase in being inhibited by capsaicin, superoxide dismutase (SOD) and N-ethyl maleimide, but differed in its sensitivity to p-chloromercuriphenylsulphonic acid (pCMBS). The NAD(P)H-induced WST-1 reduction resembled the superficial oxidase described previously, in its sensitivity to pCMBS, but differed in its suppression by capsaicin. Lucigenin was reduced by human spermatozoa in a manner that could be inhibited by SOD and stimulated by NAD(P)H or 12-myristate, 13-acetate phorbol ester. A23187 also stimulated human spermatozoa via a diphenylene iodonium sensitive pathway detectable with luminol-peroxidase but not lucigenin. Defective sperm populations recovered from the low-density region of Percoll gradients were characterized by high levels of redox activity that was only discernable with lucigenin. We conclude that human spermatozoa possess multiple plasma membrane redox systems that are involved to varying extents in the physiological control and pathological disruption of sperm function. Their distinct pharmacological profiles should significantly assist attempts to resolve and characterize these systems. PMID- 14561809 TI - Differential expression of angiopoietins 1 and 2 and their receptor Tie-2 in human endometrium. AB - Angiogenesis, the growth of new capillaries from pre-existing blood vessels, is a physiological process involved in both normal menstrual cycling and implantation of the embryo. So far, very little is known about the expression of angiopoietins, growth factors involved in angiogenesis, in human endometrium. Both angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2) are ligands for the endothelial cell-specific receptor tyrosine kinase Tie-2. In this study we determined the mRNA expression of Ang-1, Ang-2 and Tie-2 by quantitative competitive RT/(QC)-PCR (including specifically designed competitor cDNA) in biopsied human endometrium throughout the menstrual cycle. We detected the mRNA for the angiopoietins in 30 out of 32 endometrial biopsies (94%), covering early proliferative (n = 4), mid proliferative (n = 12), late proliferative (n = 3), early secretory (n = 3), mid secretory (n = 5) and late secretory (n = 3) phases. Analysis of the target/competitor ratios (QC-PCR) revealed that Ang-1 mRNA expression was significantly up-regulated (P = 0.027) during the secretory phase of the menstrual cycle. In contrast, the expression levels of both Ang-2 mRNA and Tie-2 mRNA showed only minor variations at different cycle stages. These findings were confirmed by the relative expression ratio of Ang-1 versus Ang-2 in a multiplex PCR. The expression of Ang-1, Ang-2 and Tie-2 mRNA was detected in both isolated endometrial epithelial and stromal cell fractions. Immunohistochemical localization of the proteins revealed qualitative differences in both cell type and cycle stage expression. In conclusion, the enhanced Ang-1 expression during the secretory phase might serve to stabilize the newly developed blood vessels. PMID- 14561810 TI - Large scale validation of human N-myc downstream-regulated gene (NDRG)-1 expression in endometrium during the menstrual cycle. AB - A major challenge in the comprehension of the endometrial transformations leading to the completion of each menstrual cycle in humans is in the identification of specific molecular pathways underlying these monthly turnovers. Towards this goal we compared, by the differential display technique, the relative expression of mRNA in endometrial biopsies harvested in individuals (n = 48) either at the proliferative or the secretory phase of the menstrual cycle. We isolated a cDNA fragment homologous to NDRG1 (N-myc Downstream-Regulated Gene-1) that is present in markedly higher amounts in the secretory phase. Northern blot analysis and quantitative real time PCR experiments confirmed this result in distinct cohorts of individuals (44 and 560 respectively). A closer examination of data showed that the highest mRNA levels were found during the range of 25-28 days of the uterine cycle. Consistent with the mRNA data, the temporal profile of the NDRG1 protein showed a 15-fold increase during the secretory phase, as demonstrated by using semi-quantitative dot blot analyses (n = 92). Immunohistochemical localization revealed that NDRG1 was expressed both in epithelial and stromal cells. This large scale validation of the NDRG1 mRNA and protein increase in endometrium during the secretory phase is consistent with its differentiation related function described in other tissues and its potential involvement in the window of implantation of the human endometrium, as suggested by previous chip based evidence. PMID- 14561811 TI - Human myometrial adaptation to pregnancy: cDNA microarray gene expression profiling of myometrium from non-pregnant and pregnant women. AB - The human uterus undergoes profound physiological tissue remodelling during pregnancy. In the myometrium, altered gene expression must underlie these extensive molecular and structural changes. The purpose of this study was to compare expression profiles of pregnant and non-pregnant myometrium, in order to identify genes that participate in this process. mRNA from 14 non-pregnant and four pregnant human myometrial samples were analysed using a human UniGEM V microarray with 7075 cDNA elements. A total of 602 transcripts from the microarray were up-regulated >/=2.0-fold in pregnant myometrium, with 37 transcripts up-regulated >/=4.0-fold. In contrast, eight transcripts were down regulated >/=2.0-fold in pregnancy. To ensure accurate representation of differential gene expression, Northern blot analyses using total RNA from 16 samples of non-pregnant and pregnant myometrium were used to examine mRNA levels for four of the genes that were differentially expressed by microarray analysis, namely plasminogen activator inhibitor type 1 (PAI-1), milk fat globule-EGF factor 8 protein (MFGE8), secreted frizzled-related protein 4 (sFRP4) and estrogen receptor alpha (ERalpha). On the microarray these transcripts were up regulated 7.5-fold for PAI-1 and 4.9-fold for MFGE8 in pregnant myometrium, and down-regulated 3.7-fold for sFRP4 and 2.9-fold for ERalpha in pregnancy. Northern blot analyses confirmed these changes. Our findings suggest that microarray technology is a useful tool for examining global changes in gene expression that occur as the myometrium differentiates from non-pregnant to pregnant status. Defining these changes provides new insight into the structural and functional adaptations of human myometrium during pregnancy. PMID- 14561812 TI - Clinical implications of coexpression of growth arrest-specific gene 6 and receptor tyrosine kinases Axl and Sky in human uterine leiomyoma. AB - The expression of Gas6, the protein product of the growth arrest-specific gene 6 (gas6), a member of the vitamin K-dependent protein family, and the receptor tyrosine kinases Axl and Sky and their mRNAs in uterine leiomyoma and normal uterine myometrium tissues were investigated by competitive RT-PCR-Southern blot analysis using recombinant RNA and immuno histochemical analysis respectively. There was no significant difference between the histoscores and levels of Sky mRNA in uterine leiomyoma and normal uterine myometrium, although the levels of Gas6 and Axl mRNAs in uterine leiomyoma were significantly higher than in normal uterine myometrium in each case. It is suggested that Gas6 and Axl signal transduction is aberrantly stimulated in uterine leiomyoma, possibly related to its growth. PMID- 14561813 TI - PEP-19 overexpression in human uterine leiomyoma. AB - Although uterine leiomyomas represent one of the most common neoplasms in adult women, their pathogenesis remains poorly understood. A cDNA microarray analysis was performed to search for candidate genes expressed to a greater degree in leiomyoma compared with matched myometrium. A total of 15 candidate genes was obtained; neuron-specific protein PEP-19 (Purkinje cell protein 4; PCP 4) exhibited a striking difference in expression between leiomyoma and myometrium. Although PEP-19 expression has been reported exclusively in the central nervous system, the present study demonstrated that PEP-19 is also expressed in other human organs, including prostate, kidney and uterus. To clarify the role of PEP 19 in the pathogenesis of leiomyomas, PEP-19 expression was investigated for a series of human leiomyoma, as well as normal myometrium and leiomyosarcoma. PEP 19 mRNA and protein expression were much stronger in leiomyomas compared with normal myometrium, suggesting that PEP-19 might be involved in leiomyoma pathogenesis. PMID- 14561814 TI - Endothelin-1 and macrophage colony-stimulating factor are co-localized in human amnion membrane cells and secreted into amniotic fluid. AB - We have examined the cellular localization and human amniotic fluid content of endothelin-1 (ET-1) and macrophage colony-stimulating factor (M-CSF). The study material consisted of amniotic fluid from 20 patients referred for amniocentesis, and placental samples from normal deliveries. ET-1 and M-CSF were analysed by radioimmunoassay and enzyme-linked immunosorbent assay respectively. The cellular localization of ET-1 and M-CSF in the amnion membranes was analysed by double labelling immunocytochemistry using fluorescein isothiocyanate- and Cy3-labelled secondary antibodies. Release of ET-1 and M-CSF was studied in cultured amniocytes. We found that the mean +/- SD concentrations of ET-1 and M-CSF in fetal amniotic fluid were 45.6 +/- 17.3 pmol/l (range 16.8-85.5) and 7323 +/- 3415 ng/l (range 2640-12 110) respectively. Double-labelling immunocytochemistry showed that both M-CSF and ET-1 were co-localized in the same cells to a high extent. Further analysis revealed that levels of M-CSF, but not ET-1, were significantly correlated with pregnancy length. Both M-CSF and ET-1 were released from cultured amniocytes in response to interleukin-1. These findings show that ET-1 and M-CSF are partly co-localized to specific cells in the human amniotic membrane. As both M-CSF and ET-1 were released from cultured amniocytes in vitro, this suggests that they both may be secreted into fetal amniotic fluid in vivo as well. PMID- 14561815 TI - A polymorphism in the CYP17 gene relates to the risk of recurrent pregnancy loss. AB - The CYP17 gene encodes the enzyme cytochrome P450c17alpha, which mediates both 17alpha-hydroxylase and 17,20-lyase activity in the steroid biosynthesis pathway. A T-->C polymorphism in the 5' promoter region of CYP17 has been described. To examine the association between recurrent pregnancy loss (RPL) and a polymorphism in CYP17, a case-control study of 117 cases with RPL and 164 controls was conducted. This polymorphism was investigated by PCR/restriction fragment length polymorphism using DNA from peripheral lymphocytes. The T-->C transition in the variant allele (A2) creates a new recognition site for the restriction enzyme MspA1, which permits designation of the wildtype allele (A1) and A2. Women with the A2 allele of CYP17 had an increased risk of RPL [A1/A1 genotype (reference); A1/A2 genotype: odds ratio (OR), 1.68; 95% confidence interval (CI), 0.94-3.01; A2/A2 genotype: OR, 2.37; 95% CI, 1.16-4.83; P trend, 0.016]. Additionally, there was a similar tendency for the increased risk of primary RPL [A1/A1 genotype (reference); A1/A2 genotype: OR, 2.14; 95% CI, 1.14-4.01; A2/A2 genotype: OR, 2.50; 95% CI, 1.16-5.41; P trend, 0.015]. These results suggest that possession of the A2 variant of CYP17 may predispose to an increased risk of RPL with a gene dosage effect. PMID- 14561816 TI - Immunogenicity of the soluble isoforms of HLA-G. AB - Soluble class Ib HLA-G glycoproteins synthesized in the placenta are abundant in the pregnant uterus and circulate in maternal blood throughout pregnancy. To establish immunogenicity of these proteins, we tested sera from 64 women with at least one successful pregnancy (multigravid), 21 women who had never been pregnant, and 54 males for antibodies to epitopes present on recombinant sHLA-G isoforms (sHLA-G1, sHLA-G2) derived from HLA 6.0 cDNA (HLA-G*0101 allele). By indirect enzyme-linked immunosorbent assay, antibodies to sHLA-G isoforms were identified in six sera, all from multigravid women; all other sera were negative (P = 0.0083). Immunoblots showed that two of the positive sera reacted exclusively with sHLA-G1 and -G2 whereas four reacted to both sHLA-G and pooled HLA class I antigens. To establish potential relationships between anti-sHLA-G and exposure to foreign paternal alleles (*0101, *0103, *0104, *0106), all multigravid women and their partners were genotyped. No relationship between allelic disparity and antibody production was identified. Taken together, these results indicate that (i) tolerance to HLA-G is the usual condition as antibodies to HLA-G were not detected in 91% (58/64) multigravid women, and (ii) pregnancy stimulates loss of tolerance in 9% (6/64) of multigravid women. All six women delivered healthy babies, demonstrating that maternal antibodies to epitopes on sHLA-G do not abrogate pregnancy. PMID- 14561817 TI - The L-type calcium channel C-terminus: sparking interest beyond its role in calcium-dependent inactivation. PMID- 14561818 TI - Mitochondrial formation of reactive oxygen species. AB - The reduction of oxygen to water proceeds via one electron at a time. In the mitochondrial respiratory chain, Complex IV (cytochrome oxidase) retains all partially reduced intermediates until full reduction is achieved. Other redox centres in the electron transport chain, however, may leak electrons to oxygen, partially reducing this molecule to superoxide anion (O2-*). Even though O2-* is not a strong oxidant, it is a precursor of most other reactive oxygen species, and it also becomes involved in the propagation of oxidative chain reactions. Despite the presence of various antioxidant defences, the mitochondrion appears to be the main intracellular source of these oxidants. This review describes the main mitochondrial sources of reactive species and the antioxidant defences that evolved to prevent oxidative damage in all the mitochondrial compartments. We also discuss various physiological and pathological scenarios resulting from an increased steady state concentration of mitochondrial oxidants. PMID- 14561819 TI - Subunit contributions to phosphorylation-dependent modulation of bovine rod cyclic nucleotide-gated channels. AB - Cyclic nucleotide-gated (CNG) channels in rod photoreceptors transduce a decrease in cGMP into hyperpolarization during the light response. Insulin-like growth factor-1 (IGF-1) increases light responses by increasing the cGMP sensitivity of CNG channels, an event mediated by a protein tyrosine phosphatase. Native rod CNG channels are heteromultimers, composed of three CNGA1 subunits and one CNGB1 subunit. Previous studies on heterologously expressed rod CNG channels show that a specific tyrosine in the CNGA1 subunit (Y498) is required for modulation by protein tyrosine phosphatases, protein tyrosine kinases and IGF-1. Here we show that the CNGB1 subunit contains a specific tyrosine (Y1097) that is important for modulation of heteromeric channels by tyrosine phosphorylation. Direct biochemical measurements demonstrate 32P-labelling of CNGA1Y498 and CNGB1Y1097. Replacement of either Y498 of CNGA1 or Y1097 of CNGB1 with phenylalanine reduces modulation, and removal of both tyrosines eliminates modulation. Unlike CNGA1, CNGB1 does not exhibit activity dependence of modulation by tyrosine phosphorylation. Hence both CNGA1 and CNGB1 subunits contribute to phosphorylation-dependent modulation of rod CNG channels, but the phosphorylation states of the two subunits are regulated in different ways. PMID- 14561820 TI - Long-chain acyl-CoA esters and phosphatidylinositol phosphates modulate ATP inhibition of KATP channels by the same mechanism. AB - Phosphatidylinositol phosphates (PIPs, e.g. PIP2) and long-chain acyl-CoA esters (e.g. oleoyl-CoA) are potent activators of KATP channels that are thought to link KATP channel activity to the cellular metabolism of PIPs and fatty acids. Here we show that the two types of lipid act by the same mechanism: oleoyl-CoA potently reduced the ATP sensitivity of cardiac (Kir6.2/SUR2A) and pancreatic (Kir6.2/SUR1) KATP channels in a way very similar to PIP2. Mutations (R54Q, R176A) in the C- and N-terminus of Kir6.2 that greatly reduced the PIP2 modulation of ATP sensitivity likewise reduced the modulation by oleoyl-CoA, indicating that the two lipids interact with the same site. Polyvalent cations reduced the effect of oleoyl-CoA and PIP2 on the ATP sensitivity with similar potency suggesting that electrostatic interactions are of similar importance. However, experiments with differently charged inhibitory adenosine phosphates (ATP4-, ADP3- and 2'(3')-O-(2,4,6-trinitrophenyl)adenosine 5'-monophosphate (TNP AMP2-)) and diadenosine tetraphosphate (Ap4A5-) ruled out a mechanism where oleoyl-CoA or PIP2 attenuate ATP inhibition by reducing ATP binding through electrostatic repulsion. Surprisingly, CoA (the head group of oleoyl-CoA) did not activate but inhibited KATP channels (IC50 = 265 +/- 33 muM). We provide evidence that CoA and diadenosine polyphosphates (e.g. Ap4A) are ligands of the inhibitory ATP-binding site on Kir6.2. PMID- 14561821 TI - Tight coupling of rubidium conductance and inactivation in human KCNQ1 potassium channels. AB - KCNQ1 K+ channels in humans are important for repolarization of cardiac action potentials and for K+ secretion in the inner ear. The pore-forming channel subunits form heteromeric complexes with small regulatory subunits of the KCNE family, in particular with KCNE1 to form channels that conduct a slow delayed rectifier K+ current, IKs. This association leads to alteration of biophysical properties, including a slowing of activation, a suppression of inactivation and an increase of the apparent single-channel conductance. In addition, inward Rb+ currents conducted by homomeric KCNQ1 channels are about threefold larger than K+ currents, whereas heteromeric KCNQ1-KCNE1 channels have smaller inward Rb+ currents compared to K+ currents. We determined inactivation properties and compared K+ vs. Rb+ inward currents for channels formed by co-assembly of KCNQ1 with KCNE1, KCNE3 and KCNE5, and for homomeric KCNQ1 channels with point mutations in the pore helix S5 or S6 transmembrane domains. Several of the channels with point mutations eliminated the apparent inactivation of KCNQ1, as described previously (Seebohm et al. 2001). We found that the extent of inactivation and the ratio of Rb+/K+ currents were positively correlated. Since the effect of Rb+ on the current size has been shown previously to be related to a fast 'flickery' process, our results suggest that inactivation of KCNQ1 channels is related to a fast flicker of the open channel. A kinetic model incorporating two open states, no explicit inactivated state and a fast flicker that is different for the two open states is able to account for the apparent inactivation and the correlation of inactivation and large Rb+ currents. We conclude that an association between KCNQ1 and KCNE subunits or removal of inactivation by mutation of KCNQ1 stabilizes the open conformation of the pore principally by altering an interaction between the pore helix and the selectivity filter and with S5/S6 domains. PMID- 14561822 TI - Conditional protein phosphorylation regulates BK channel activity in rat cerebellar Purkinje neurons. AB - Large conductance calcium- and voltage-activated potassium (BK) channels are widely expressed in the mammalian central nervous system. Although the activity of BK channels in endocrine and vascular cells is regulated by protein kinases and phosphatases associated with the channel complex, direct evidence for such modulation in neurons is largely lacking. Single-channel analysis from inside-out patches isolated from the soma of dissociated rat cerebellar Purkinje neurons demonstrated that the activity of BK channels is regulated by multiple endogenous protein kinases and protein phosphatases in the membrane patch. The majority of BK channels were non-inactivating and displayed a 'low' activity phenotype determined at +40 mV and 1 muM intracellular free calcium. These channels were activated by cAMP-dependent protein kinase (PKA) associated with the patch and the extent of PKA activation was limited by an opposing endogenous type 2A-like protein phosphatase (PP2A). Importantly, PKA activation was dependent upon the prior phosphorylation status of the BK channel complex dynamically controlled by protein kinase C (PKC) and protein phosphatase 1 (PP1). In contrast, Purkinje cells also displayed a low proportion of non-inactivating BK channels with a 'high' activity under the same recording conditions and these channels were inhibited by endogenous PKA. Our data suggest that: (1) multiple endogenous protein kinases and phosphatases functionally couple to the BK channel complex to allow conditional modulation of BK channel activity in neurons, and (2) native, phenotypically distinct, neuronal BK channels are differentially sensitive to PKA dependent phosphorylation. PMID- 14561823 TI - Mitochondrial function in intact skeletal muscle fibres of creatine kinase deficient mice. AB - Creatine kinase (CK) has a central role in skeletal muscle, acting as a fast energy buffer and shuttle between sites of energy production (mitochondria) and consumption (cross-bridges and ion pumps). Unexpectedly, isolated fast-twitch skeletal muscle cells of mice deficient in both cytosolic and mitochondrial CK (CK-/-) are highly fatigue resistant during stimulation protocols that stress aerobic metabolism. We have now studied different aspects of mitochondrial function in CK-/- skeletal muscle. Intact, single fibres of flexor digitorum brevis (FDB) muscles were fatigued by repeated tetanic stimulation (70 Hz, 350 ms duration, duty cycle 0.14). Under control conditions, CK-/- FDB fibres were more fatigue resistant than wild-type fibres. However, after mitochondrial inhibition with cyanide, force declined markedly faster in CK-/- fibres than in wild-type fibres. The rapid force decline in CK-/- fibres was not due to decreased myoplasmic [Ca2+] during tetani (measured with indo-1), which in these fibres remained virtually constant during fatigue in the presence of cyanide. Intact, single fibres of highly oxidative soleus muscles were fatigued by repeated tetani (50 Hz, 500 ms duration, duty cycle 0.5). All CK-/- soleus fibres tested (n = 9) produced > 40 % force at the end of the fatiguing stimulation period (500 tetani), whereas force fell to < 40 % before 500 tetani in two of three wild-type fibres. Mitochondrial [Ca2+] (measured with rhod-2 and confocal microscopy) increased during repeated tetanic stimulation in CK-/- but not in wild-type FDB fibres. In conclusion, mitochondria and energy shuttling operate effectively in CK-/- fibres and this is associated with an increase in mitochondrial [Ca2+]. PMID- 14561824 TI - Modulation of RhoA-Rho kinase-mediated Ca2+ sensitization of rabbit myometrium during pregnancy - role of Rnd3. AB - During pregnancy, the uterus undergoes major functional and structural remodelling. It is well known that during the major part of pregnancy, the myometrium normally remains relatively quiescent but is able to generate powerful contractions at the time of parturition. However, the intracellular molecular events regulating myometrial contractility during pregnancy still remain poorly understood. We applied differential gene expression screening using cDNA array technology to probe myometrium samples from non-pregnant and mid-pregnant (15 days) rabbits. Among the differentially expressed genes, the farnesylated small G protein of the Rho family, Rnd3, was found to be upregulated (3.6-fold) at mid pregnancy. Upregulation of Rnd3 was confirmed at the protein level by a 3.4-fold increase in Rnd3 expression in mid-pregnant myometrium. Measurements of contractile properties of beta-escin permeabilized smooth muscle strips revealed that the upregulation of Rnd3 correlated with an inhibition of RhoA-Rho kinase mediated Ca2+ sensitization at mid-pregnancy. Treatment of muscle strips from mid pregnant myometrium with the farnesyl-transferase inhibitor manumycin A (10 muM) led to the recovery of RhoA-Rho kinase-dependent Ca2+ sensitization. At late pregnancy (31 days), upregulation of RhoA and Rho kinase expression was associated with an increase in Ca2+ sensitivity of contractile proteins that was inhibited by the Rho kinase inhibitor Y-27632 (10 muM). These data thus demonstrate the time-dependent regulation of the RhoA-Rho kinase-mediated Ca2+ sensitization during the course of pregnancy. The depression of this mechanism at mid-pregnancy followed by its constitutive activation near term is associated with a co-ordinated modulation of Rnd3, RhoA and Rho kinase expression. The RhoA Rho kinase signalling pathway and its regulators might thus represent potential targets for the development of new treatments for pre-term labour. PMID- 14561825 TI - L-type Ca2+ channels serve as a sensor of the SR Ca2+ for tuning the efficacy of Ca2+-induced Ca2+ release in rat ventricular myocytes. AB - In cardiac excitation-contraction coupling, Ca2+-induced Ca2+ release (CICR) from ryanodine receptors (RyRs), triggered by Ca2+ entry through the nearby L-type Ca2+ channel, induces Ca2+-dependent inactivation (CDI) of the Ca2+ channel. Aiming at elucidating the physiological role of CDI produced by CICR (CICR dependent CDI), we investigated the contribution of the CICR-dependent CDI to action potential (AP) waveform and the amount of Ca2+-influx through Ca2+ channels during AP in rat ventricular myocytes. The elimination of the CICR dependent CDI, by depletion of the SR Ca2+ with thapsigargin, significantly prolonged AP duration (APD). APD changed in parallel with the magnitude of CICR during the recovery of the SR Ca2+ content after transient depletion by caffeine. Such CICR-dependent change of APD persisted under the highly Ca2+ buffered condition where the Ca2+ signalling was restricted to nanoscale domains. Blockers of the Ca2+-dependent Cl- channel or the BK channel did not affect AP waveform. The amount of Ca2+-influx through Ca2+ channels during the SR-depleted type AP waveform, measured in the SR-depleted myocyte, was increased by 40 % over that during the SR-intact type AP waveform measured in the SR-intact myocyte. The protein kinase A stimulation further enhanced the Ca2+-influx during AP under the SR-depleted condition to 70 % of that under the SR-intact condition. These results indicate that the CICR-dependent CDI of L-type Ca2+ channels, under control of the privileged cross-signalling between L-type Ca2+ channels and RyRs, play important roles for monitoring and tuning the SR Ca2+ content via changes of AP waveform and the amount of Ca2+-influx during AP in ventricular myocytes. PMID- 14561826 TI - Apical and basal neurones isolated from the mouse vomeronasal organ differ for voltage-dependent currents. AB - The mammalian vomeronasal organ (VNO) contains specialized neurones that transduce the chemical information related to pheromones into discharge of action potentials to the brain. Molecular and biochemical studies have shown that specific components of the pheromonal transduction systems are segregated into two distinct subsets of vomeronasal neurones: apical neurones and basal neurones. However, it is still unknown whether these neuronal subsets also differ in other functional characteristics, such as their membrane properties. We addressed this issue by studying the electrophysiological properties of vomeronasal neurones isolated from mouse VNO. We used the patch-clamp technique to examine both the passive membrane properties and the voltage-gated Na+, K+ and Ca2+ currents. Apical neurones were distinguished from basal ones by the length of their dendrites and by their distinct immunoreactivity for the putative pheromone receptor V2R2. The analysis of passive properties revealed that there were no significant differences between the two neuronal subsets. Also, apical neurones were similar to basal neurones in their biophysical and pharmacological properties of voltage-gated Na+ and K+ currents. However, we found that the density of Na+ currents was about 2-3 times greater in apical neurones than in basal neurones. Consistently, in situ hybridization analysis revealed a higher expression of the Na+ channel subtype III in apical neurones than in basal ones. In contrast, basal neurones were endowed with Ca2+ currents (T-type) of greater magnitude than apical neurones. Our findings indicate that apical and basal neurones in the VNO exhibit distinct electrical properties. This might have a profound effect on the sensory processes occurring in the VNO during pheromone detection. PMID- 14561827 TI - Modulation of Ca2+ signalling in rat atrial myocytes: possible role of the alpha1C carboxyl terminal. AB - Ca2+ influx through L-type Cav1.2 (alpha1C) Ca2+ channels is a critical step in the activation of cardiac ryanodine receptors (RyRs) and release of Ca2+ via Ca2+ induced Ca2+ release(CICR). The released Ca2+, in turn, is the dominant determinant of inactivation of the Ca2+ current (ICa) and termination of release. Although Ca2+ cross-signalling is mediated by high Ca2+ fluxes in the microdomains of alpha1C-RyR complexes, ICa-gated Ca2+ cross-signalling is surprisingly resistant to intracellular Ca2+ buffering and has steeply voltage dependent gain, inconsistent with a strict CICR mechanism, suggesting the existence of additional regulatory step(s). To explore the possible regulatory role of the carboxyl (C)-terminal tail of alpha1C in modulating Ca2+ signalling, we tested the effects of introducing two alpha1C C-terminal peptides, LA (1571 1599) and K (1617-1636) on the central alpha1C-unassociated Ca2+-release sites of atrial myocytes, using rapid (240 Hz) two-dimensional confocal Ca2+ imaging. The frequency of spontaneously activating central sparks increased by approximately fourfold on dialysing LA- but not K-peptide into myocytes voltage-clamped at -80 mV. The rate but not the magnitude of caffeine (10 mM)-triggered central Ca2+ release was significantly accelerated by LA- but not K-peptide. Individual Ca2+ spark size and flux were larger in LA- but not in K-peptide-dialysed myocytes. Although LA-peptide did not change the amplitude or inactivation kinetics of ICa, LA-peptide did strongly enhance the central Ca2+ transients triggered by ICa at 30 mV (small ICa) but not at +20 mV (large ICa). In contrast, K-peptide had no effect on either ICa or the local Ca2+ transients. LA-peptide with a deleted calmodulin-binding region (LM1-peptide) had no significant effects on the central spark frequency but suppressed spontaneous spark frequency in the periphery. Our results indicate that the calmodulin-binding LA motif of the alpha1C C-terminal tail may sensitize the RyRs, thereby increasing their open probability and providing for both the voltage-dependence of CICR and the higher frequency of spark occurrence in the periphery of atrial myocytes where the native alpha1C-RyR complexes are intact. PMID- 14561829 TI - Unmyelinated axons in the rat hippocampus hyperpolarize and activate an H current when spike frequency exceeds 1 Hz. AB - The mammalian cortex is densely populated by extensively branching, thin, unmyelinated axons that form en passant synapses. Some thin axons in the peripheral nervous system hyperpolarize if action potential frequency exceeds 1-5 Hz. To test the hypothesis that cortical axons also show activity-induced hyperpolarization, we recorded extracellularly from individual CA3 pyramidal neurons while activating their axon with trains consisting of 30 electrical stimuli. Synaptic excitation was blocked by kynurenic acid. We observed a positive correlation between stimulation strength and the number of consecutive axonal stimuli that resulted in soma spikes, suggesting that the threshold increased as a function of the number of spikes. During trains without response failures there was always a cumulative increase in the soma response latency. Intermittent failures, however, decreased the latency of the subsequent response. At frequencies of > 1 Hz, the threshold and latency increases were enhanced by blocking the hyperpolarization-activated H current (Ih)by applying the specific Ih blocker ZD7288 (25 microM) or 2 mM Cs+. Under these conditions, response failures occurred after 15-25 stimuli, independent of the stimulation strength. Adding GABA receptor blockers (saclofen and bicuculline) and a blocker of metabotropic glutamate receptors did not change the activity-induced latency increase in recordings of the compound action potential. We interpret these results as an activity-induced hyperpolarization that is partly counteracted by Ih. Such a hyperpolarization may influence transmitter release and the conduction reliability of these axons. PMID- 14561828 TI - Gadolinium reduces short-term stretch-induced muscle damage in isolated mdx mouse muscle fibres. AB - Duchenne muscular dystrophy is a lethal muscle disease caused by absence of the protein dystrophin which is part of a glycoprotein complex located on the intracellular surface of the surface membrane. The precise function of dystrophin and the reason why its absence causes severe muscle damage are unclear. Stretch induced muscle damage is well recognised in normal muscle and is more severe in muscles from animals lacking dystrophin (mdx mice). It has been proposed that stretch-induced damage underlies the progression of damage in muscular dystrophy. In the present study we confirm that single fibres from mdx muscle are more susceptible to stretch-induced damage and show that there is an associated rise in intracellular sodium concentration ([Na+]i) which is greater than in wild-type mice. We show that this rise in [Na+]i can be prevented by Gd3+, which is an established blocker of stretch-activated channels. mdx fibres have a higher than normal resting [Na+]i and this is also reduced by Gd3+. If Gd3+ is applied over the period in which [Na+]i rises following stretched contraction, it prevents one component of the reduced force. The other component of reduced force is caused by inhomogeneity of sarcomeres and can be minimised by stretching the muscle to its new optimum length. These experiments show that part of the short-term damage caused by stretch in mdx fibres can be prevented by blocking stretch-activated channels. PMID- 14561830 TI - Transforming growth factor-beta1 reduces megalin- and cubilin-mediated endocytosis of albumin in proximal-tubule-derived opossum kidney cells. AB - Transforming growth factor (TGF)-beta1 is a member of a superfamily of multifunctional cytokines involved in several pathological processes of the kidney, including fibrogenesis, apoptosis and epithelial-mesenchymal transition. These events lead to tubulointerstitial fibrosis and glomerulosclerosis. Less is known about TGF-beta1-induced alterations of cell function. An important function of proximal tubular cells is reabsorption of filtered proteins, including albumin, via megalin-cubilin-dependent receptor-mediated endocytosis. In this study we used a well established cell culture model (proximal-tubule-derived opossum kidney (OK) cells) in order to test the hypothesis that TGF-beta1 reduces megalin-cubilin-mediated endocytosis. Previously we have shown that albumin endocytosis in OK cells is mediated by megalin/cubulin. TGF-beta1 led to a time- and dose-dependent downregulation of megalin-cubilin-mediated endocytosis without affecting two other transport systems tested. Binding, internalization and intracellular trafficking of the ligand albumin were affected. Decreased binding resulted from reduced cubilin and megalin expression in the 200 000 g membrane fraction. The underlying mechanism of TGF-beta1 action does not involve mitogen activated protein kinases, protein kinase C or A, or reactive oxygen species. In contrast, TGF-beta1-induced downregulation of megalin-cubilin-mediated endocytosis was sensitive to inhibition of translation and transcription and was preceded by Smad2 and 3 phosphorylation. Dominant negative Smad2/3 constructs prevented the effect of TGF-beta1. In conclusion our data indicate that enhanced levels of TGF-beta1 occurring in various nephropathies can lead to downregulation of megalin-cubilin-dependent endocytosis. Probably, TGF-beta1 leads to Smad2- and Smad3-dependent expression of negative regulators of receptor-mediated endocytosis. PMID- 14561832 TI - The effect of ageing and immobilization on structure and function of human skeletal muscle fibres. AB - Biopsy samples were taken from vastus lateralis muscle of seven young (YO, age 30.2 +/- 2.2 years), and seven elderly (EL, age 72.7 +/- 2.3 years) subjects and two elderly subjects whose right leg had been immobilized for 3.5 months (EL-IMM, ages 70 and 75). The following main parameters were studied: (1) myosin heavy chain (MHC) isoform distribution of the samples, determined by SDS-PAGE; (2) cross-sectional area (CSA), specific force (Po/CSA) and maximum shortening velocity (Vo) of a large population (n = 593) of single skinned muscle fibres, classified on the basis of MHC isoform composition determined by SDS-PAGE; (3) actin sliding velocity (Vf) on pure myosin isoforms determined by in vitro motility assays; (4) myosin concentration in single fibres determined by quantitative SDS-PAGE. MHC isoform distribution was shifted towards fast isoforms in EL and to a larger extent in EL-IMM. In EL and, more consistently, in EL-IMM we observed a higher percentage of hybrid fibres than in YO, and noted the presence of MHC-neonatal and of unusual hybrid fibres containing more than two MHC isoforms. Po/CSA significantly decreased in type 1 and 2A fibres in the order YO EL EL-IMM. Vo of type 1 and 2A fibres was significantly lower in EL and higher in EL-IMM than in YO, i.e. immobilization more than counteracted the age dependent decrease in Vo. The latter phenomenon was not observed for Vf. Vf on myosin 1 was lower in both EL and EL-IMM than in YO. Vf on myosin 2X was lower in EL than in YO, and a similar trend was observed for myosin 2A. Myosin concentration decreased in type 1 and 2A fibres in the order YO EL EL-IMM and was linearly related to the Po/CSA values of corresponding fibre types from the same subjects. The experiments suggest that (1) myosin concentration is a major determinant of the lower Po/CSA of single fibres in ageing and especially following immobilization and (2) ageing is associated with lower Vo of single fibres due to changes in the properties of myosin itself, whereas immobilization is associated with higher Vo in the absence of a change in myosin function. PMID- 14561831 TI - Ca2+-activated K+ (BK) channel inactivation contributes to spike broadening during repetitive firing in the rat lateral amygdala. AB - In many neurons, trains of action potentials show frequency-dependent broadening. This broadening results from the voltage-dependent inactivation of K+ currents that contribute to action potential repolarisation. In different neuronal cell types these K+ currents have been shown to be either slowly inactivating delayed rectifier type currents or rapidly inactivating A-type voltage-gated K+ currents. Recent findings show that inactivation of a Ca2+-dependent K+ current, mediated by large conductance BK-type channels, also contributes to spike broadening. Here, using whole-cell recordings in acute slices, we examine spike broadening in lateral amygdala projection neurons. Spike broadening is frequency dependent and is reversed by brief hyperpolarisations. This broadening is reduced by blockade of voltage-gated Ca2+ channels and BK channels. In contrast, broadening is not blocked by high concentrations of 4-aminopyridine (4-AP) or alpha-dendrotoxin. We conclude that while inactivation of BK-type Ca2+-activated K+ channels contributes to spike broadening in lateral amygdala neurons, inactivation of another as yet unidentified outward current also plays a role. PMID- 14561833 TI - An ID-like current that is downregulated by Ca2+ modulates information coding at CA3-CA3 synapses in the rat hippocampus. AB - Voltage-gated K+ channels localised on presynaptic nerve terminals control information coding by modulating presynaptic firing and synaptic efficacy in target neurones. We found that at CA3-CA3 connections in hippocampal slice cultures, a fast-activating, slowly inactivating K+ conductance similar to the so called delay current (ID) is responsible for the delayed appearance of the first spike upon membrane depolarisation, for action potential repolarisation and for modulation of transmitter release. The ID-like current was downregulated by intracellular Ca2+, as indicated by the increased delay in the appearance of the first action potential following either the block of Ca2+ flux through voltage dependent Ca2+ channels with Cd2+ or replacement of the bathing solution with one devoid of Ca2+. In both cases, this effect was reversed by blocking this conductance with a low concentration of 4-aminopyridine (4-AP, 10-50 muM). Application of 4-AP shortened the delay to the first spike generation, prevented the effect of Cd2+ and increased the spike duration. The earlier appearance of the first action potential was also observed in the presence of dendrotoxin-1 (100 nM). In voltage-clamp experiments larger currents were recorded in the absence of extracellular Ca2+, thus confirming the downregulation of the ID-like current by Ca2+ due to the positive shift of its inactivation. Spike broadening was associated with an enhancement of synaptic efficacy in target neurones, as assessed by the increase in EPSC amplitude and in the percentage of successes. Moreover, in the presence of 4-AP, EPSCs appeared with a longer latency and were more scattered. This conductance is therefore crucial for setting the timing and strength of synaptic transmission at CA3-CA3 connections. It is conceivable that switching off ID by increasing intracellular Ca2+ following activity-dependent processes may facilitate network synchronisation and crosstalk between CA3 pyramidal cells, leading to seizure activity. PMID- 14561834 TI - Membrane properties of type II spiral ganglion neurones identified in a neonatal rat cochlear slice. AB - Neuro-anatomical studies in the mammalian cochlea have previously identified a subpopulation of approximately 5 % of primary auditory neurones, designated type II spiral ganglion neurones (sgnII). These neurones project to outer hair cells and their supporting cells, within the 'cochlear amplifier' region. Physiological characterization of sgnII has proven elusive. Whole-cell patch clamp of spiral ganglion neurones in P7-P10 rat cochlear slices provided functional characterization of sgnII, identified by biocytin or Lucifer yellow labelling of their peripheral neurite projections (outer spiral fibres) subsequent to electrophysiological characterisation. SgnII terminal fields comprised multiple outer hair cells and supporting cells, located up to 370 mum basal to their soma. SgnII firing properties were defined by rapidly inactivating A-type-like potassium currents that suppress burst firing of action potentials. Type I spiral ganglion neurones (sgnI), had shorter radial projections to single inner hair cells and exhibited larger potassium currents with faster activation and slower inactivation kinetics, compatible with the high temporal firing fidelity seen in auditory nerve coding. Based on these findings, sgnII may be identified in future by the A-type current. Glutamate-gated somatic currents in sgnII were more potentiated by cyclothiazide than those in sgnI, suggesting differential AMPA receptor expression. ATP-activated desensitising inward currents were comparable in sgn II and sgnI. These data support a role for sgnII in providing integrated afferent feedback from the cochlear amplifier. PMID- 14561835 TI - Electrogram prolongation and nifedipine-suppressible ventricular arrhythmias in mice following targeted disruption of KCNE1. AB - Mutations in KCNE1, the gene encoding the beta subunit of the slowly activating delayed rectifier potassium current (IKs) channel protein, may lead to the long QT syndrome (LQTS), a condition associated with enhanced arrhythmogenesis. Mice with homozygous deletion of the coding sequence of KCNE1 have inner ear defects strikingly similar to those seen in the corresponding human condition. The present study demonstrated and assessed the mechanism of ventricular arrhythmias in Langendorff-perfused whole heart preparations from homozygous KCNE1-/- mice compared to wild-type mice of the same age. The effects of programmed electrical stimulation with decremental pacing from the basal right ventricular epicardial surface upon electrogram waveforms recorded from the basal left ventricle were assessed and quantified using techniques of paced electrogram fractionation analysis for the first time in an experimental system. All KCNE1-/-(n = 10) but not wild-type (n = 14) mouse hearts empirically demonstrated marked pacing induced ventricular arrhythmogenicity. This correlated with significant increases in electrogram dispersion, consistent with a wider spread in conduction velocities, in parallel with clinical findings from LQTS patients with potassium channel mutations. In contrast, introduction of 100 nM isoprenaline induced arrhythmogenicity in both KCNE1-/- (n = 7) and wild-type (n = 6) hearts during pacing. Furthermore, pretreatment with 1 muM nifedipine exerted a strong anti arrhythmic effect in the KCNE1-/- hearts (n = 12) that persisted even in the presence of 100 nM isoprenaline (n = 6). Our findings associate KCNE1-/- with an arrhythmogenic phenotype that shows an increased dispersion of conduction velocities, and whose initiation is prevented by nifedipine, a finding that in turn may have therapeutic applications in conditions such as LQTS. PMID- 14561837 TI - Hypoxic preconditioning enhances renal superoxide dismutase levels in rats. AB - Renal ischaemia releases reactive oxygen species (ROS) in the kidneys. We hypothesized that the kidneys are more resistant to the insult of ROS in chronically hypoxic rats. We thus compared rats kept at sea level (SL) and those that had been adapted to hypoxia (hypoxia adapted, HA) by exposure to an altitude of 5500 m in an altitude chamber for 15 h day-1 for 4 weeks. Xanthine (X, 0.75 mg kg-1) and xanthine oxidase (XO, 24.8 mU kg-1) were injected intrarenally. A lucigenin-enhanced chemiluminescence method was employed to detect the amount of free radicals in renal venous blood samples and on the kidney surface. In the renal venous blood samples, 26.05 (+/- 4.36) x 104 and 10.98 (+/- 1.79) x 104 counts were detected in the SL and HA rats, respectively, after X-XO treatment; these figures were significantly different. On the kidney surface of the SL rats, the free radical count amounted to 12.77 (+/- 1.64) x 104, while that in the HA rats was 8.47 (+/- 0.42) x 104; these figures were also significantly different. There was a significant increase in urine volume and urinary excretion of Na+, K+ and protein after X-XO administration in both groups of rats. However, the effect was greater for the SL rats than for the HA rats. The lipid peroxidation of the kidneys was not significantly different in the two groups of rats. Finally, we found that the activity of superoxide dismutase (SOD) and SOD mRNA were higher in the renal tissue of HA rats. We conclude that the renal response to free radicals is attenuated after chronic hypoxia in rats, and that SOD might play an important role in protecting HA rats from oxidative stress. PMID- 14561836 TI - Substance P presynaptically depresses the transmission of sensory input to bronchopulmonary neurons in the guinea pig nucleus tractus solitarii. AB - Substance P modulates the reflex regulation of respiratory function by its actions both peripherally and in the CNS, particularly in the nucleus tractus solitarii (NTS), the first central site for synaptic contact of the lung and airway afferent fibres. There is considerable evidence that the actions of substance P in the NTS augment respiratory reflex output, but the precise effects on synaptic transmission have not yet been determined. Therefore, we determined the effects of substance P on synaptic transmission at the first central synapses by using whole-cell voltage clamping in an NTS slice preparation. Studies were performed on second-order neurons in the slice anatomically identified as receiving monosynaptic input from sensory nerves in the lungs and airways. This was done by the fluorescent labelling of terminal boutons after 1,1'-dioctadecyl 3,3,3',3'-tetra-methylindocarbo-cyanine perchlorate (DiI) was applied via tracheal instillation. Substance P (1.0, 0.3 and 0.1 microM) significantly decreased the amplitude of excitatory postsynaptic currents (eEPSCs) evoked by stimulation of the tractus solitarius, in a concentration-dependent manner. The decrease was accompanied by an increase in the paired-pulse ratio of two consecutive eEPSCs, and a decrease in the frequency, but not the amplitude, of spontaneous EPSCs and miniature EPSCs, findings consistent with a presynaptic site of action. The effects were consistently and significantly attenuated by a neurokinin-1 (NK1) receptor antagonist (SR140333, 3 muM). The data suggest a new site of action for substance P in the NTS (NK1 receptors on the central terminals of sensory fibres) and a new mechanism (depression of synaptic transmission) for regulating respiratory reflex function. PMID- 14561838 TI - Hypothalamic regulation of pancreatic secretion is mediated by central cholinergic pathways in the rat. AB - The vago-vagal reflex plays an important role in mediating pancreatic secretion evoked by cholecystokinin and non-cholecystokinin-dependent luminal factors. We hypothesize that the vago-vagal reflex mediating pancreatic secretion in the rat is under central control and regulated by cholinergic pathways in the hypothalamus. To test this hypothesis, we demonstrated that chronic decerebration decreased basal pancreatic enzyme secretion from 318 +/- 12 to 233 +/- 9 mg h-1 and reduced the net increase in pancreatic secretion stimulated by intraduodenal infusion of 5 % peptone and hypertonic NaCl by 54 % and 45 %, respectively. Intracerebroventricular administration of methscopolamine (MSCP, 50 nmol (5 mul) 1), a blood-brain barrier-impermeant cholinergic muscarinic receptor antagonist, evoked results similar to those achieved by chronic decerebration. To localize the sites of action, we demonstrated that microinjection of MSCP (20 nmol) into the lateral hypothalamic nucleus or the paraventricular nucleus resulted in inhibition of both basal pancreatic protein secretion and luminally stimulated pancreatic secretion by 48 % and 52 %, respectively. Intracerebroventricular injection of hemicholinium-3 at doses known to deplete the endogenous ACh store produced similar inhibitory results. In addition, microinjection of ACh (5 pmol) or the muscarinic M1 receptor agonist McN-A-343 (30 ng) into the lateral hypothalamic nucleus increased pancreatic secretion over basal levels by 46 % and 40 %, respectively. Selective lesions of lateral septal cholinergic neurons decreased basal pancreatic secretion and inhibited peptone-induced pancreatic secretion by 30 %. Destruction of the lateral parabrachial nucleus produced a 44 % inhibition of peptone-induced pancreatic section. Finally, microinjection of glutamate into the lateral septum or the lateral parabrachial nucleus stimulated vagal pancreatic efferent nerve firings from a basal level of 0 +/- 0.5 impulses (30 s)-1 to 4.5 +/- 0.5 and 14 +/- 2 impulses (30 s)-1, respectively, and pancreatic protein output increased 50 % and 84 % over basal levels. Administration of MSCP to the paraventricular nucleus eliminated these effects. These observations suggest that cholinergic neurons of the lateral septum and lateral parabrachial nucleus regulate pancreatic secretion. Further, cholinergic input from the lateral parabrachial nucleus to the hypothalamus plays a major role in the modulation of vagal pancreatic efferent nerve activity and pancreatic secretion evoked by the vago-vagal reflex. PMID- 14561840 TI - Iron deficiency during pregnancy affects postnatal blood pressure in the rat. AB - Iron (Fe) deficiency anaemia during pregnancy results in an increased risk of perinatal mortality and morbidity and is a significant factor for increased risk of disease in later life. Consequently we have developed a rat model to study the relationship between maternal Fe deficiency and postnatal growth and blood pressure in the offspring. Weanlings were fed a control or Fe-deficient diet prior to and throughout pregnancy. At term, all pups were cross-fostered to control fed dams and weaned onto control diet. At birth, pups from deficient dams had a greater mortality rate, were smaller and had reduced haematocrit and liver Fe levels. They also had larger hearts, smaller kidneys and spleens and unchanged livers (relative organ weight). The pups grew normally. At 6 weeks, male pups from deficient dams had a higher and females a lower blood pressure than their normal counterparts. At 10 and 16 weeks, blood pressure in the males from deficient dams was still raised and in the females was now greater than controls. The haematocrit was lower in males throughout the 16 weeks and in females until 10 weeks of age. There was no significant difference in the offsprings' liver Fe stores at 6, 10 or 16 weeks. Duodenal Fe uptake in both the Fe-deficient mother and newborn offspring was significantly increased. By cross-fostering, we have eliminated confounding factors, such as maternal anaemia during lactation and show, unequivocally, that prenatal nutrition is critical for the development of normal postnatal function. PMID- 14561839 TI - Stimulation of proteinase-activated receptor 2 excites jejunal afferent nerves in anaesthetised rats. AB - Proteinase-activated receptor 2 (PAR2) is a receptor for mast cell tryptase and trypsins and might participate in brain-gut communication. However, evidence that PAR2 activation can lead to afferent impulse generation is lacking. To address this issue, we examined the sensitivity of jejunal afferent nerves to a hexapeptide agonist of PAR2, SLIGRL-NH2, and the modulation of the resulting response to treatment with drugs and vagotomy. Multiunit recordings of jejunal afferent activity were made using extracellular recording techniques in anaesthetised male rats. SLIGRL-NH2 (0.001-1 mg kg-1, I.V.) increased jejunal afferent firing and intrajejunal pressure. The reverse peptide sequence (1 mg kg 1, I.V.), which does not stimulate PAR2, was inactive. Naproxen (10 mg kg-1, I.V.), but not a cocktail of omega-conotoxins GVIA and SVIB (each at 25 mug kg-1, I.V.), curtailed both the afferent response and the intrajejunal pressure rise elicited by the PAR2 agonist. Although neither treatment modulated the peak magnitude of the afferent firing, they each altered the intestinal motor response, unmasking an initial inhibitory component. Nifedipine (1 mg kg-1, I.V.) reduced the peak magnitude of the afferent nerve discharge and abolished the initial rise in intrajejunal pressure produced by SLIGRL-NH2. Vagotomy did not significantly influence the magnitude of the afferent response to the PAR2 agonist, which involves a contribution from capsaicin-sensitive fibres. In conclusion, intravenous administration of SLIGRL-NH2 evokes complex activation of predominantly spinally projecting extrinsic intestinal afferent nerves, an effect that involves both direct and indirect mechanisms. PMID- 14561841 TI - Evaluation of the limiting acuity of coincidence detection in nucleus laminaris of the chicken. AB - The localization of sounds requires the detection of very brief inter-aural time differences (ITDs). In birds, ITDs are first encoded in neurons of the nucleus laminaris (NL) through the precise coincidence of binaural synaptic inputs. We examined the effects of temperature on acuity of coincidence detection in chick NL, by utilizing whole-cell and cell-attached recording techniques in brain slices while applying electrical stimuli bilaterally to axonal projections from the nucleus magnocellularis to NL. The precision of coincidence detection was measured as a time window, corresponding to the time interval that gave the half maximum spiking probability. Acuity improved with the elevation of recording temperature, and at 40 degrees C, the avian body temperature, the time window was 0.38 ms. Although all synaptic events were briefer at higher temperature, the duration of EPSPs were equivalent to or faster than that of EPSCs at 40 degrees C. Activation of low-threshold K+ currents by a slight membrane depolarization during an EPSP was responsible for this EPSP acceleration. EPSPs were prolonged following inhibition of low-threshold K+ currents by dendrotoxin (40 nM) or hyperpolarization-activated cation currents by Cs+ (3 mM). The EPSP time course had a strong positive correlation with the sharpness of coincidence detection. The limiting value of the time window (0.16 ms), calculated from the estimated EPSP time course, was narrow enough to explain the acuity of ITD detection at NL in vivo. PMID- 14561842 TI - The effect of intrafetal infusion of metyrapone on arterial blood pressure and on the arterial blood pressure response to angiotensin II in the sheep fetus during late gestation. AB - While the impact of exogenous glucocorticoids on the fetal cardiovascular system has been well defined, relatively few studies have characterised the role of endogenous fetal glucocorticoids in the regulation of arterial blood pressure (BP) during late gestation. We have therefore infused metyrapone, an inhibitor of cortisol biosynthesis, into fetal sheep from 125 days gestation (when fetal cortisol concentrations are low) and from 137 days gestation (when fetal cortisol concentrations are increasing) and measured fetal plasma cortisol, 11 desoxycortisol and ACTH, fetal systolic, diastolic and mean arterial BP, heart rate, and the fetal BP responses to increasing doses of angiotensin II (AII). At 125 days gestation, there was a significant increase in fetal plasma ACTH and 11 desoxycortisol by 24 h after (+24 h) the start of the metyrapone infusion, and plasma cortisol concentrations were not different at +24 h when compared with pre infusion values. Whilst the initial fall in circulating cortisol concentrations may have been transient, systolic, diastolic and mean arterial BP were ~5-6 mmHg lower (P < 0.05) in metyrapone- than in vehicle-infused fetuses at 24-48 h after the start of the infusion. When metyrapone was infused from 137/138 days gestation, there was a significant decrease in plasma cortisol concentrations by +6 h, which was followed by an increase back to pre-infusion values. While cortisol concentrations decreased, there was no change in fetal mean arterial BP during the first 24 h after the start of metyrapone infusion. Mean fetal arterial BP values at 137-139 days gestation were not different in fetuses that had been infused with either vehicle or metyrapone from 125 days gestation or with metyrapone from 137/138 days gestation. At 137-139 days gestation, however, arterial BP responses to increasing doses of AII were significantly blunted in fetuses that had been infused with metyrapone from 125 days gestation, when compared with fetuses that had been infused with metyrapone from 137/138 days gestation or with vehicle from 125 days gestation. The dissociation of the gestational age increase in arterial BP and the effects of intrafetal AII on fetal arterial BP indicates that increase in fetal BP with gestational age is not entirely a result of an increased vascular responsiveness to endogenous AII. Furthermore there may be a critical window during late gestation when the actions of cortisol contribute to the development of vascular responsiveness to AII. PMID- 14561843 TI - Influences of hydration on post-exercise cardiovascular control in humans. AB - Dehydration is known to decrease orthostatic tolerance and cause tachycardia, but little is known about the cardiovascular control mechanisms involved. To test the hypothesis that arterial baroreflex sensitivity increases during exercise-induced dehydration, we assessed arterial baroreflex responsiveness in 13 healthy subjects (protocol 1) at baseline (PRE-EX) and 1 h after (EX-DEH) 90 min of exercise to cause dehydration, and after subsequent intravenous rehydration with saline (EX-REH). Six of these subjects were studied a second time (protocol 2) with intravenous saline during exercise to prevent dehydration. We measured heart rate, central venous pressure and arterial pressure during all trials, and muscle sympathetic nerve activity (MSNA) during the post-exercise trials. Baroreflex responses were assessed using sequential boluses of nitroprusside and phenylephrine (modified Oxford technique). After exercise in protocol 1 (EX-DEH), resting blood pressure was decreased and resting heart rate was increased. Cardiac baroreflex gain, assessed as the responsiveness of heart rate or R-R interval to changes in systolic pressure, was diminished in the EX-DEH condition (9.17 +/- 1.06 ms mmHg-1 vs. PRE-EX: 18.68 +/- 2.22 ms mmHg-1, P < 0.05). Saline infusion after exercise did not alter the increase in HR post-exercise or the decrease in baroreflex gain (EX-REH: 10.20 +/- 1.43 ms mmHg-1; P > 0.10 vs. EX DEH). Saline infusion during exercise (protocol 2) resulted in less of a post exercise decrease in blood pressure and a smaller change in cardiac baroreflex sensitivity. Saline infusion caused a decrease in MSNA in protocol 1. We conclude that exercise-induced dehydration causes post-exercise changes in the baroreflex control of blood pressure that may contribute to, rather than offset, orthostatic intolerance. PMID- 14561844 TI - Motor unit recruitment and derecruitment induced by brief increase in contraction amplitude of the human trapezius muscle. AB - The activity pattern of low-threshold human trapezius motor units was examined in response to brief, voluntary increases in contraction amplitude ('EMG pulse') superimposed on a constant contraction at 4-7 % of the surface electromyographic (EMG) response at maximal voluntary contraction (4-7 % EMGmax). EMG pulses at 15 20 % EMGmax were superimposed every minute on contractions of 5, 10, or 30 min duration. A quadrifilar fine-wire electrode recorded single motor unit activity and a surface electrode recorded simultaneously the surface EMG signal. Low threshold motor units recruited at the start of the contraction were observed to stop firing while motor units of higher recruitment threshold stayed active. Derecruitment of a motor unit coincided with the end of an EMG pulse. The lowest threshold motor units showed only brief silent periods. Some motor units with recruitment threshold up to 5 % EMGmax higher than the constant contraction level were recruited during an EMG pulse and kept firing throughout the contraction. Following an EMG pulse, there was a marked reduction in motor unit firing rates upon return of the surface EMG signal to the constant contraction level, outlasting the EMG pulse by 4 s on average. The reduction in firing rates may serve as a trigger to induce derecruitment. We speculate that the silent periods following derecruitment may be due to deactivation of non-inactivating inward current ('plateau potentials'). The firing behaviour of trapezius motor units in these experiments may thus illustrate a mechanism and a control strategy to reduce fatigue of motor units with sustained activity patterns. PMID- 14561846 TI - Pharmacogenetic considerations in diseases of cardiac ion channels. AB - Phenotypic variation within a species arises from differences in genetic makeup between individuals. This inherent diversity empowers the species as a whole to explore and expand into new environmental niches and also to survive new stressors within an ever-changing environment. Paradoxically, one class of stressors currently challenging the human population is therapeutic drugs: medications designed to combat disease are often associated with a host of nonspecific side effects. Following earlier studies of the involvement of some cardiac ion currents in unwanted drug interactions, recent reports have identified not only the ion channel subunits involved but also a range of mutations and single nucleotide polymorphisms in ion channel genes that predispose to both drug-induced and familial cardiac arrhythmia. The tendency for individuals harboring specific, often common, gene variants to succumb to life threatening cardiac arrhythmia, and the contribution of other factors such as drug interaction to disease etiology in these cases, are discussed here together with potential pharmacogenetic strategies for arrhythmia circumvention and therapy. PMID- 14561845 TI - The time course of the motoneurone afterhyperpolarization is related to motor unit twitch speed in human skeletal muscle. AB - The relationship between the electrophysiological properties of motoneurones and their muscle units has been established in animal models. A functionally significant relationship exists whereby motoneurones with long post-spike afterhyperpolarizations (AHPs) innervate slow contracting muscle units. The purpose of this study was to determine whether the time course of the AHP as measured by its time constant is associated with the contractile properties of its muscle unit in humans. Using an intramuscular fine wire electrode, 46 motor units were recorded in eight subjects as they held a low force contraction of the first dorsal interosseus muscle for approximately 10 min. By applying a recently validated transform to the interspike interval histogram, the mean voltage versus time trajectory of the motoneurone AHP was determined. Spike-triggered averaging was used to extract the muscle unit twitch from the whole muscle force with strict control over force variability and motor unit discharge rate (interspike intervals between 120 and 200 ms). The AHP time constant was positively correlated to the time to half-force decay (rho = 0.36, P < 0.05) and twitch duration (rho = 0.57, P < 0.001); however, time to peak force failed to reach significance (rho = 0.27, P < 0.07). These results suggest that a similar functional relationship exists in humans between the motoneurone AHP and the muscle unit contractile properties. PMID- 14561847 TI - Pharmacokinetics and pharmacodynamics of methylecgonidine, a crack cocaine pyrolyzate. AB - Methylecgonidine is formed from cocaine base when smoked and has been identified in biological fluids of crack smokers. Ecgonidine, a metabolite of methylecgonidine formed via esterase activity, also has been identified in similar samples collected from crack smokers. Methylecgonidine and ecgonidine can be used as biomarkers to differentiate smoking from cocaine use via other routes of administration. We determined the pharmacokinetic properties of methylecgonidine and ecgonidine in sheep after intravenous administration of methylecgonidine at doses of 3.0, 5.6, and 10.0 mg/kg using gas chromatography mass spectrometric assays. Methylecgonidine clears quickly from blood with a half life of 18 to 21 min, whereas ecgonidine has a longer half-life of 94 to 137 min. Because ecgonidine clears more slowly, it may be a more effective biomarker of cocaine smoking. The cardiovascular stimulant effects of cocaine contrast with reported in vitro muscarinic agonist effects of methylecgonidine, decreasing contractility and stimulating nitric oxide production in cardiac cells and tissues. To test the hypothesis that methylecgonidine produces cardiovascular effects in vivo consistent with muscarinic agonism, methylecgonidine was administered to sheep intravenously (0.1-3.0 mg/kg) while monitoring heart rate and blood pressure. Significant hypotension and tachycardia occurred in all three sheep. Two of the three sheep demonstrated mild bradycardia 3 to 5 min after methylecgonidine injection. Intravenous pretreatment with atropine methyl bromide (15 microg/kg) antagonized methylecgonidine-induced hypotension in all three sheep, supporting the hypothesis that methylecgonidine acts as a muscarinic agonist in vivo. PMID- 14561848 TI - Neuronal and astroglial responses to the serotonin and norepinephrine neurotoxin: 1-methyl-4-(2'-aminophenyl)-1,2,3,6-tetrahydropyridine. AB - 1-Methyl-4-(2'-aminophenyl)-1,2,3,6-tetrahydropyridine (2'-NH2-MPTP) causes long term loss of forebrain serotonin (5-HT) and norepinephrine (NE) and consequently, is unlike 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and its other 2' analogs that primarily deplete striatal dopamine (DA). In the present investigation into the acute effects of 2'-NH2-MPTP in mice, profound decreases in cortical and hippocampal 5-HT and NE to 10 to 40% of control were observed as early as 30 min post-treatment and lasted throughout the ensuing 21 days. Striatal DA was decreased to 60 to 80% of control during the first 48 h but returned to normal by 72 h. Reactive gliosis, which occurs in response to neurodegeneration was not evident by immunocytochemistry but was detected by enzyme-linked immunosorbent assay, where glial fibrillary acidic protein (GFAP) was increased to 130% of control in cortex, hippocampus, and brain stem 48 to 72 h post-treatment. To explore the possibility that 5-HT modulates the astrocytic response to injury, 2'-NH2-MPTP was used to damage 5-HT axons 2 weeks before administration of the potent DA neurotoxin 1-methyl-4-(2'-methylphenyl)-1,2,3,6 tetrahydropyridine (2'-CH3-MPTP). Despite a 90% decrement in striatal DA in 2' NH2-MPTP/2'-CH3-MPTP-treated mice, increases in GFAP were attenuated compared to mice treated with 2'-CH3-MPTP alone. Thus, 2'-NH2-MPTP causes severe and immediate decrements in 5-HT and NE in frontal cortex and hippocampus, yet induces a modest GFAP response compared with other MPTP analogs that have their primary effect on DA. These results demonstrate the importance of obtaining quantitative assessments of GFAP to detect astroglial responses associated with selective damage to neurotransmitter systems with low-density innervation and suggest that serotonin may facilitate the astrocytic response to striatal injury. PMID- 14561849 TI - Unique regulation profile of prostaglandin e1 on adhesion molecule expression and cytokine production in human peripheral blood mononuclear cells. AB - In the present study, we examined the effects of prostaglandin E1 (PGE1) on the expression of intercellular adhesion molecule (ICAM)-1, B7.1, B7.2, CD40, and CD40 ligand (CD40L) on peripheral blood mononuclear cells (PBMC) using fluorescence-activated cell sorting analysis as well as its effects on cytokine production using enzyme-linked immunosorbent assay. Whereas no inhibitor of spontaneous expression of adhesion molecules was reported, we found that PGE1 inhibited spontaneous ICAM-1, B7.2, and CD40 expression on monocytes in a concentration-dependent manner but had no effect on the expression of B7.1 and CD40L. Although interleukin (IL)-18 induced the expression of ICAM-1, B7.2, CD40, and CD40L, PGE1 prevented IL-18-induced expression of ICAM-1, B7.2, and CD40. We examined the involvement of five subtypes of PGE1 receptors (IP, EP1, EP2, EP3, and EP4) in the effect of PGE1 on the expression of these adhesion molecules using subtype-specific agonists. Among EP receptor agonists, EP2 and EP4 receptor agonists inhibited IL-18-elicited ICAM-1, B7.2, and CD40 expression. ONO-1301 (IP receptor agonist) prevented the expression of ICAM-1, B7.2, and CD40 regardless of the presence of IL-18 with the same potency as PGE1. The effect of a combination of ONO-1301 and 11-deoxy (D)-PGE1 (EP2/EP4 receptor agonist) on ICAM 1, B7.2, and CD40 expression mimicked that of PGE1. Moreover, PGE1 inhibited the production of IL-12 and interferon-gamma in PBMC in the presence and absence of IL-18, whereas PGE1 induced IL-10 production. In conclusion, IP receptor and EP2/EP4 receptor play an important role in the action of PGE1 on the expression of adhesion molecules on monocytes and cytokine production. PMID- 14561850 TI - p38 MAPK inhibitors ameliorate target organ damage in hypertension: Part 2. Improved renal function as assessed by dynamic contrast-enhanced magnetic resonance imaging. AB - Recent evidence suggests p38 mitogen-activated protein kinase (MAPK) signal transduction plays an important role in the pathogenesis of progressive renal disease. Using dynamic contrast enhanced magnetic resonance imaging (MRI), we evaluated chronic treatment with a p38 MAPK inhibitor, trans-1-(4 hydroxycyclohexyl)-4-(4-fluorophenyl-methoxypyridimidin-4-yl)imidazole (SB 239063), on renal function in a hypertension model of progressing renal dysfunction. Spontaneously hypertensive-stroke prone rats were placed on a high salt/fat diet (SFD) or maintained on normal chow diet (ND). SFD animals with albuminuria at 4 to 8 weeks (> or =10 mg/day inclusion criteria), were randomized into p38 MAPK inhibitor treatment (SB-239063, 1200 ppm in diet) or vehicle groups. The progression of blood pressure and albuminuria during the treatment period (approximately 6 weeks) was decreased by 12 and 60%, respectively, in the SFD + SB-239063 versus SFD control group. Renal perfusion and filtration were assessed by in vivo MRI at the end of the study. Relative cortical perfusion was increased in the SFD + SB-239063 group compared with the SFD control group as reflected by a 29% decrease in time to peak of contrast agent in the cortex. Additionally, the regional renal glomerular filtration rate index (Kcl) was increased by 39% in the SFD + SB-239063 versus SFD control group and was normalized to the ND control group. Greater functional heterogeneity was observed in the SFD control versus SFD + SB-239063 or ND control group. All alterations of renal function were supported by histopathological findings. In conclusion, chronic treatment with a p38 MAPK inhibitor, SB-239063, attenuates functional and structural renal degeneration in a hypertensive model of established renal dysfunction. PMID- 14561851 TI - p38 MAPK inhibitors ameliorate target organ damage in hypertension: Part 1. p38 MAPK-dependent endothelial dysfunction and hypertension. AB - Numerous mediators, believed to play a role in endothelial dysfunction (e.g., neurohormones, cytokines, hypoxia, and stretch), have been shown to activate p38 mitogen-activated protein kinase (MAPK) in a variety of cell types. The purpose of the present study was to examine the regulation of p38 MAPK in endothelium and its role in endothelial dysfunction and salt sensitivity. In cultured human umbilical vein endothelial cells (HUVECs), tumor necrosis factor-alpha and lipopolysaccharide increased phosphorylation of p38 MAPK (P-p38 MAPK) and increased ICAM-1 expression. Preincubation with highly selective p38 MAPK inhibitors, 1-(1,3-dihydroxyprop-2-yl)-4-(4-fluorophenyl)-5-[2-phenoxypyrimidin-4 yl] imidazole (SB-239063AN) or SB-239063, dose dependently reduced intercellular adhesion molecule-1 expression in HUVECs. In spontaneously hypertensive-stroke prone rats (SHR-SP), P-p38 MAPK was localized by immunohistochemistry to the aortic endothelium and adventitia but was undetectable in aortae from normotensive rats. Introduction of a salt/fat diet (SFD) to the SHR-SP strain induced endothelial dysfunction (ex vivo vascular reactivity analysis), albuminuria, and an increase in blood pressure within 4 weeks. Chronic dietary dosing (approx. 100 mg/kg/day) with SB-239063AN inhibited the SFD diet-induced hypertension. In addition, delayed treatment also significantly improved survival and restored nitric oxide-mediated endothelium-dependent relaxation in SFD-SHR SPs with established endothelial dysfunction. These results suggest an important role for p38 MAPK in endothelial inflammation and dysfunction as well as providing the first evidence for p38 MAPK-dependent hypertension. PMID- 14561852 TI - Beneficial effects of heme oxygenase-1 up-regulation in the development of experimental inflammation induced by zymosan. AB - Heme oxygenase-1 (HO-1) is part of the integrated response to oxidative stress. This enzyme may exert anti-inflammatory effects in some animal models, although the precise mechanisms are not fully understood. We have examined the role of HO 1 in the inflammatory response induced by zymosan in the mouse air pouch. Zymosan administration induced HO-1 protein expression in leukocytes migrating to exudates, with maximal levels in the late phase of this response (24-48 h). This was accompanied by ferritin induction and bilirubin accumulation, indicating that this enzyme is active in our model. HO-1 expression by zymosan treatment was partly reduced by aminoguanidine, suggesting the participation of endogenous nitric oxide in the mechanisms leading to HO-1 synthesis in the zymosan-injected mouse air pouch. Up-regulation of HO-1 by hemin administration resulted in inhibition of nitric-oxide synthase-2 activity, cellular infiltration into the air pouch exudate, and plasmatic exudation. Leukotriene B4 levels in exudates were significantly decreased in the early phase of this response (4 h), whereas interleukin-1beta and tumor necrosis factor-alpha were inhibited at all time points. Inhibition of HO-1 activity by zinc protoporphyrin IX prevented most of the effects caused by hemin administration. Our results indicate that HO-1 exerts anti-inflammatory effects on the response to zymosan in the mouse air pouch and support a role for this enzyme in the modulation of inflammatory processes. PMID- 14561853 TI - Resolution of sensory ambiguities for gaze stabilization requires a second neural integrator. AB - The ability to simultaneously move in the world and maintain stable visual perception depends critically on the contribution of vestibulo-ocular reflexes (VORs) to gaze stabilization. It is traditionally believed that semicircular canal signals drive compensatory responses to rotational head disturbances (rotational VOR), whereas otolith signals compensate for translational movements [translational VOR (TVOR)]. However, a sensory ambiguity exists because otolith afferents are activated similarly during head translations and reorientations relative to gravity (i.e., tilts). Extra-otolith cues are, therefore, necessary to ensure that dynamic head tilts do not elicit a TVOR. To investigate how extra otolith signals contribute, we characterized the temporal and viewing distance dependent properties of a TVOR elicited in the absence of a lateral acceleration stimulus to the otoliths during combined translational/rotational motion. We show that, in addition to otolith signals, angular head position signals derived by integrating sensory canal information drive the TVOR. A physiological basis for these results is proposed in a model with two distinct integration steps. Upstream of the well known oculomotor velocity-to-position neural integrator, the model incorporates a separate integration element that could represent the "velocity storage integrator," whose functional role in the oculomotor system has so far remained controversial. We propose that a key functional purpose of the velocity storage network is to temporally integrate semicircular canal signals, so that they may be used to extract translation information from ambiguous otolith afferent signals in the natural and functionally relevant bandwidth of head movements. PMID- 14561854 TI - Axonal regeneration through regions of chondroitin sulfate proteoglycan deposition after spinal cord injury: a balance of permissiveness and inhibition. AB - Increased expression of certain extracellular matrix (ECM) molecules after CNS injury is believed to restrict axonal regeneration. The chondroitin sulfate proteoglycans (CSPGs) are one such class of ECM molecules that inhibit neurite outgrowth in vitro and are upregulated after CNS injury. We examined growth responses of several classes of axons to this inhibitory environment in the presence of a cellular fibroblast bridge in a spinal cord lesion site and after a growth factor stimulus at the lesion site (fibroblasts genetically modified to secrete NGF). Immunohistochemical analysis showed dense labeling of the CSPGs NG2, brevican, neurocan, versican, and phosphacan at the host-lesion interface after spinal cord injury (SCI). Furthermore, robust expression of NG2, and to a lesser extent versican, was also observed throughout grafts of control and NGF secreting fibroblasts. Despite this inhibitory milieu, several axonal classes penetrated control fibroblast grafts, including dorsal column sensory, rubrospinal, and nociceptive axons. Axon growth was amplified more in the presence of NGF-secreting grafts. Confocal microscopy demonstrated that axon growth was, paradoxically, preferentially associated with NG2-rich substrates in both graft types. NG2 expression also increased after sciatic nerve injury, wherein axons successfully regenerate. Cellular sources of NG2 in SCI and peripheral nerve lesion sites included Schwann cells and endothelial cells. Notably, these same cellular sources in lesion sites produced the cell adhesion molecules L1 and laminin, and these molecules all colocalized. Thus, axons grow along substrates coexpressing both inhibitory and permissive molecules, suggesting that regeneration is successful when local permissive signals balance and exceed inhibitory signals. PMID- 14561855 TI - Suppression of adult neurogenesis impairs olfactory learning and memory in an adult insect. AB - Although adult neurogenesis has now been demonstrated in many different species, the functional role of newborn neurons still remains unclear. In the house cricket, a cluster of neuroblasts, located in the main associative center of the insect brain, keeps producing new interneurons throughout the animal's life. Here we address the functional significance of adult neurogenesis by specific suppression of neuroblast proliferation using gamma irradiation of the insect's head and by examining the impact on the insect's learning ability. Forty gray irradiation performed on the first day of adult life massively suppressed neuroblasts and their progeny without inducing any noticeable side effect. We developed a new operant conditioning paradigm especially designed for crickets: the "escape paradigm." Using olfactory cues, visual cues, or both, crickets had to choose between two holes, one allowing them to escape and the other leading to a trap. Crickets lacking adult neurogenesis exhibited delayed learning when olfactory cues alone were used. Furthermore, retention 24 hr after conditioning was strongly impaired in irradiated crickets. By contrast, when visual cues instead of olfactory ones were provided, performance of irradiated insects was only slightly affected; when both olfactory and visual cues were present, their performance was not different from that of controls. From these results, it can be postulated that newborn neurons participate in the processing of olfactory information required for complex operant conditioning. PMID- 14561856 TI - Cholinergic activity enhances hippocampal long-term potentiation in CA1 during walking in rats. AB - Long-term potentiation (LTP) at the basal-dendritic synapses of CA1 pyramidal cells was induced by a single 200 Hz stimulation train (0.5-1 sec duration) in freely behaving rats during one of four behavioral states: awake-immobility (IMM), walking, slow-wave sleep (SWS), and rapid eye movement sleep (REM). Field EPSPs generated by basal-dendritic excitation of CA1 were recorded before and up to 1 d after the tetanus. After a tetanus during any behavioral state, basal dendritic LTP was >170% of the baseline for the first hour after the tetanus and decayed to approximately 120% 1 d after. LTP induced during walking was significantly larger than that induced during IMM, SWS, or REM, which had similar LTP magnitudes. To test the hypothesis that septohippocampal cholinergic activity enhanced LTP during walking as compared with IMM, rats were either pretreated with muscarinic cholinergic antagonist scopolamine or injected with IgG192 saporin in the medial septum to selectively lesion cholinergic septohippocampal neurons. Pretreatment with scopolamine decreased the LTP induced during walking but did not affect that induced during IMM, such that the difference between the LTP induced during walking and IMM was abolished after scopolamine. Rats injected with IgG192-saporin showed no difference in the LTP induced during walking and IMM, and scopolamine did not reduce the LTP during walking. In contrast, sham lesion rats showed larger LTP induced during walking than IMM, and the LTP induced during walking was attenuated by scopolamine. This is the first demonstration of an enhancement of hippocampal LTP by physiologically activated cholinergic inputs. PMID- 14561857 TI - Involvement of the olfactory tubercle in cocaine reward: intracranial self administration studies. AB - Cocaine has multiple actions and multiple sites of action in the brain. Evidence from pharmacological studies indicates that it is the ability of cocaine to block dopamine uptake and elevate extracellular dopamine concentrations, and thus increase dopaminergic receptor activation, that makes cocaine rewarding. Lesion studies have implicated the nucleus accumbens (the dorsal portion of the "ventral striatum") as the probable site of the rewarding action of the drug. However, the drug is only marginally self-administered into this site. We now report that cocaine (60 or 200 mm in 75 nl/infusion) is readily self-administered into the olfactory tubercle, the most ventral portion of the ventral striatum. Cocaine (200 mm) was self-administered marginally into the accumbens shell but not into the core, dorsal striatum, or ventral pallidum. In addition, cocaine injections (200 mm in 300 nl) into the tubercle but not the shell or ventral pallidum induced conditioned place preference. Rewarding effects of cocaine in the tubercle were blocked by coadministration of dopamine D1 or D2 antagonists (1 mm SCH 23390 or 3 mm raclopride) and were not mimicked by injections of the local anesthetic procaine (800 mm). In conclusion, the tubercle plays a critical role in mediating rewarding action of cocaine. PMID- 14561858 TI - An intact intermediate filament network is required for collateral sprouting of small diameter nerve fibers. AB - Expression of the intermediate filament (IF) protein peripherin is initiated during development at the time of axonal extension and increases during regeneration of nerve fibers. To test whether the IF network is essential for neuron process outgrowth in the mature organism in vivo, we disrupted endogenous peripherin IF in small-sized dorsal root ganglion (DRG) neurons in transgenic mice via expression of a mutant peripherin transgene under control of peripherin gene regulatory sequences. Anatomical and functional analyses showed that these neurons send peripheral and central axonal projections to correct targets, express correct neuropeptides, and mediate acute pain responses normally. However, disruption of IF significantly impaired the ability of uninjured small sized DRG neurons to sprout collateral axons into adjacent denervated skin, indicating a critical role for intact IF in plasticity, specifically in compensatory nociceptive nerve sprouting. PMID- 14561859 TI - Altered neuronal excitability in cerebellar granule cells of mice lacking calretinin. AB - Calcium-binding proteins such as calretinin are abundantly expressed in distinctive patterns in the CNS, but their physiological function remains poorly understood. Calretinin is expressed in cerebellar granule cells, which provide the major excitatory input to Purkinje cells through parallel fibers. Calretinin deficient mice exhibit dramatic alterations in motor coordination and Purkinje cell firing recorded in vivo through unknown mechanisms. In the present study, we used patch-clamp recording techniques in acute slice preparation to investigate the effect of a null mutation of the calretinin gene on the intrinsic electroresponsiveness of cerebellar granule cells at a mature developmental stage. Calretinin-deficient granule cells exhibit faster action potentials and generate repetitive spike discharge showing an enhanced frequency increase with injected currents. These alterations disappear when 0.15 mm of the exogenous fast calcium buffer BAPTA is infused in the cytosol to restore the calcium-buffering capacity. A proposed mathematical model demonstrates that the observed alterations of granule cell excitability can be explained by a decreased cytosolic calcium-buffering capacity resulting from the absence of calretinin. This result suggests that calcium-binding proteins modulate intrinsic neuronal excitability and may therefore play a role in information processing in the CNS. PMID- 14561860 TI - Correlation between olfactory receptor cell type and function in the channel catfish. AB - The olfactory epithelium of fish contains three intermingled types of olfactory receptor neurons (ORNs): ciliated, microvillous, and crypt. The present experiments were undertaken to test whether the different types of ORNs respond to different classes of odorants via different families of receptor molecules and G-proteins corresponding to the morphology of the ORN. In catfish, ciliated ORNs express OR-type receptors and Galpha(olf). Microvillous ORNs are heterogeneous, with many expressing Galpha(q)/11, whereas crypt ORNs express Galpha(o). Retrograde tracing experiments show that ciliated ORNs project predominantly to regions of the olfactory bulb (OB) that respond to bile salts (medial) and amino acids (ventral) (Nikonov and Caprio, 2001). In contrast, microvillous ORNs project almost entirely to the dorsal surface of the OB, where responses to nucleotides (posterior OB) and amino acids (anterior OB) predominate. These anatomical findings are consistent with our pharmacological results showing that forskolin (which interferes with Galpha(olf)/cAMP signaling) blocks responses to bile salts and markedly reduces responses to amino acids. Conversely, U-73122 and U-73343 (which interfere with Galpha(q)/11/phospholipase C signaling) diminish amino acid responses but leave bile salt and nucleotide responses essentially unchanged. In summary, our results indicate that bile salt odorants are detected predominantly by ciliated ORNs relying on the Galpha(olf)/cAMP transduction cascade. Nucleotides are detected by microvillous ORNs using neither Galpha(olf)/cAMP nor Galpha(q)/11/PLC cascades. Finally, amino acid odorants activate both ciliated and microvillous ORNs but via different transduction pathways in the two types of cells. PMID- 14561861 TI - Activity-dependent plasticity of transmitter release from nerve terminals in rat fast and slow muscles. AB - Neuromuscular junctions (NMJs) on fast and slow muscle fibers display different transmitter release characteristics that appear well adapted to the different patterns of nerve impulses that they transmit in vivo. Here, we ask whether the release properties of such NMJs, termed fast and slow, can be transformed by chronic nerve stimulation. In young adult rats, nerve impulse conduction in the sciatic nerve was blocked by TTX, and the nerve to the fast extensor digitorum longus (EDL) or the slow soleus (SOL) muscle stimulated directly below the block with slow (20 Hz for 10 sec every 30 sec) or fast (150 Hz for 1 sec every 60 sec) stimulus patterns, respectively. After 3-4 weeks, originally fast EDL-NMJs and slow SOL-NMJs had become almost fully transformed to slow and fast NMJs, respectively, with respect to maintenance of transmitter release during tonic 20 Hz stimulation in vitro and ratio of quantal content to vesicle pool size. TTX block alone had no such transforming effect. Vesicle recycle time was unaffected by the stimulation, whereas initial quantal content and vesicle pool size were reduced (by 49% and 57% in EDL and 33% and 67% in SOL). Muscle fiber diameter also declined (by 49% in EDL and 33% in SOL vs 46% in unstimulated SOL; unstimulated EDL was not examined). We conclude that fast and slow NMJs display marked plasticity by being able to adapt important release characteristics to the impulse patterns imposed on them. PMID- 14561862 TI - Selective adaptation in networks of cortical neurons. AB - A key property of neural systems is their ability to adapt selectively to stimuli with different features. Using multisite electrical recordings from networks of cortical neurons developing ex vivo, we show that neurons adapt selectively to different stimuli invading the network. We focus on selective adaptation to frequent and rare stimuli; networks were stimulated at two sites with two different stimulus frequencies. When both stimuli were presented within the same period, neurons in the network attenuated their responsiveness to the more frequent input, whereas their responsiveness to the rarely delivered stimuli showed a marked average increase. The amplification of the response to rare stimuli required the presence of the other, more frequent stimulation source. By contrast, the decreased response to the frequent stimuli occurred regardless of the presence of the rare stimuli. Analysis of the response of single units suggests that both of these effects are caused by changes in synaptic transmission. By using synaptic blockers, we find that the increased responsiveness to the rarely stimulated site depends specifically on fast GABAergic transmission. Thus, excitatory synaptic depression, the inhibitory sub network, and their balance play an active role in generating selective gain control. The observation that selective adaptation arises naturally in a network of cortical neurons developing ex vivo indicates that this is an inherent feature of spontaneously organizing cortical networks. PMID- 14561863 TI - Two distinct subpopulations of nestin-positive cells in adult mouse dentate gyrus. AB - Neurogenesis in the dentate gyrus of the adult mammalian hippocampus has been proven in a series of studies, but the differentiation process toward newborn neurons is still unclear. In addition to the immunohistochemical study, electrophysiological membrane recordings of precursor cells could provide an alternative view to address this differentiation process. In this study, we performed green fluorescent protein (GFP)-guided selective recordings of nestin positive progenitor cells in adult dentate gyrus by means of nestin-promoter GFP transgenic mice, because nestin is a typical marker for precursor cells in the adult dentate gyrus. The patch-clamp recordings clearly demonstrated the presence of two distinct subpopulations (type I and type II) of nestin-positive cells. Type I cells had a lower input resistance value of 77.1 M(Omega) (geometric mean), and their radial processes were stained with anti-glial fibrillary acidic protein antibody. On the other hand, type II nestin-positive cells had a higher input resistance value of 2110 MOmega and expressed voltage-dependent sodium current. In most cases, type II cells were stained with anti-polysialylated neural cell adhesion molecule. Taken together with a bromodeoxyuridine pulse chase analysis, our results may reflect a rapid and dynamic cell conversion of nestin-positive progenitor, from type I to type II, at an early stage of adult neurogenesis in the dentate gyrus. PMID- 14561864 TI - Aberrant formation of glutamate receptor complexes in hippocampal neurons of mice lacking the GluR2 AMPA receptor subunit. AB - The number and type of receptors present at the postsynaptic membrane determine the response to the neurotransmitter released from the presynaptic terminal. Because most neurons receive multiple and distinct synaptic inputs and contain several different subtypes of receptors stimulated by the same neurotransmitter, the assembly and trafficking of receptors in neurons is a complex process involving many levels of regulation. To investigate the mechanism that neurons use to regulate the assembly of receptor subunits, we studied a GluR2 knock-out mouse. GluR2 is a critical subunit that controls calcium permeability of AMPA receptors and is present in most native AMPA receptors. Our data indicate that in the absence of GluR2, aberrant receptor complexes composed of GluR1 and GluR3 are formed in the hippocampus, and that there is an increased number of homomeric GluR1 and GluR3 receptors. We also show that these homomeric and heteromeric receptors are less efficiently expressed at the synapse. Our results show that GluR2 plays a critical role in controlling the assembly of AMPA receptors, and that the assembly of subunits may reflect the affinity of one subunit for another or the stability of intermediates in the assembly process. Therefore, GluR1 may have a greater preference for GluR2 than it does for GluR3. PMID- 14561865 TI - Biphasic effects of cannabinoids on acetylcholine release in the hippocampus: site and mechanism of action. AB - Cannabinoids have been shown to critically modulate cholinergic neurotransmission in the hippocampus, yet opposing effects of cannabinoid receptor 1 (CB1R) agonists on hippocampal synaptic acetylcholine (ACh) efflux have been reported. This study shows that administration of a synthetic CB1R agonist results in a biphasic, dose-dependent, effect on hippocampal ACh: a low (0.5 mg/kg, i.p.) and a high (5 mg/kg, i.p) dose of WIN55,212-2 induces a transient stimulation and a prolonged inhibition of hippocampal ACh efflux, respectively. Both effects of WIN55,212-2 are mediated through CB1 receptors coupled to Gi but involve different neuroanatomical sites. Thus, intrahippocampal infusion of the CB1R antagonist SR141716A or pertussis toxin blocked the inhibition of hippocampal ACh release induced by the high dose of WIN55,212-2, but was without effect on the stimulatory action of the low dose. In contrast, this latter effect was blocked by SR141716A or pertussis toxin infused, in dual microdialysis experiments, in the septum, in which the majority of cholinergic cell bodies projecting to the hippocampus reside. The stimulatory and inhibitory effects of WIN55,212-2 on hippocampal ACh involve dopamine D1 and D2 receptor activation, respectively, given that pretreatment with D1 and D2 receptor antagonists prevents the respective actions of WIN55,212-2. We propose that the in vivo observed biphasic effects of CB1R agonists on hippocampal ACh release result from a differential, functional association of anatomicaly distinct subpopulations of CB1-Gi coupled receptors to neurotransmitter systems that have opposing effects on ACh release. This concept could provide a theoretical framework to understand endocannabinoids as state-dependent modulators of neuronal activity. PMID- 14561866 TI - ZNRF proteins constitute a family of presynaptic E3 ubiquitin ligases. AB - Protein ubiquitination has been implicated recently in neural development, plasticity, and degeneration. We previously identified ZNRF1/nin283, a protein with a unique, evolutionarily conserved C-terminal domain containing a juxtaposed zinc finger/RING finger combination. Here we describe the identification of a closely related protein, ZNRF2, thus defining a novel family of ZNRF E3 ubiquitin ligases. Both ZNRF1 and ZNRF2 have E3 ubiquitin ligase activity and are highly expressed in the nervous system, particularly during development. In neurons, ZNRF proteins are located in different compartments within the presynaptic terminal: ZNRF1 is associated with synaptic vesicle membranes, whereas ZNRF2 is present in presynaptic plasma membranes. Mutant ZNRF proteins with a disrupted RING finger, a domain necessary for their E3 function, can each inhibit Ca2+ dependent exocytosis in PC12 cells. These data suggest that ZNRF proteins play a role in the establishment and maintenance of neuronal transmission and plasticity via their ubiquitin ligase activity. PMID- 14561867 TI - Hyperdopaminergic mutant mice have higher "wanting" but not "liking" for sweet rewards. AB - What is the role of dopamine in natural rewards? A genetic mutant approach was taken to examine the consequences of elevated synaptic dopamine on (1) spontaneous food and water intake, (2) incentive motivation and learning to obtain a palatable sweet reward in a runway task, and (3) affective "liking" reactions elicited by the taste of sucrose. A dopamine transporter (DAT) knockdown mutation that preserves only 10% of normal DAT, and therefore causes mutant mice to have 70% elevated levels of synaptic dopamine, was used to identify dopamine effects on food intake and reward. We found that hyperdopaminergic DAT knockdown mutant mice have higher food and water intake. In a runway task, they demonstrated enhanced acquisition and greater incentive performance for a sweet reward. Hyperdopaminergic mutant mice leave the start box more quickly than wild-type mice, require fewer trials to learn, pause less often in the runway, resist distractions better, and proceed more directly to the goal. Those observations suggest that hyperdopaminergic mutant mice attribute greater incentive salience ("wanting") to a sweet reward in the runway test. But sucrose taste fails to elicit higher orofacial "liking" reactions from mutant mice in an affective taste reactivity test. These results indicate that chronically elevated extracellular dopamine facilitates "wanting" and learning of an incentive motivation task for a sweet reward, but elevated dopamine does not increase "liking" reactions to the hedonic impact of sweet tastes. PMID- 14561868 TI - Forebrain-specific neuronal inhibition of nuclear factor-kappaB activity leads to loss of neuroprotection. AB - The transcription factor Rel/nuclear factor (NF)-kappaB is known for its fundamental role in regulating immune and inflammatory responses. In the brain, constitutive NF-kappaB activity has been detected exclusively in neurons, and a large diversity of stimuli have been reported to induce NF-kappaB activity. Yet the function of this transcription factor in the nervous system remains unclear, and its role in neuroprotection or neurodegeneration is open to debate. Recently it was suggested that kappaB-driven gene expression in neurons is controlled by Sp1-like factors. To clarify such controversy, we have characterized here a novel mouse model in which the entire NF-kappaB-dependent transcriptional response is abolished in the forebrain. Calcium-calmodulin-dependent kinase II alpha promoter driven tetracycline transactivator was used for regulated expression of a transdominant negative mutant of inhibitor kappaBalpha (super-repressor) together with a green fluorescent protein tracer. Inhibition of expression of a kappaB dependent lacZ transgene was shown in triple transgenic mice, which correlated with the loss of kappaB-specific DNA binding. In transgenic organotypic hippocampal slice cultures, expression of the super-repressor led to strong cell death after neurotoxic insults. These data demonstrate for the first time that neuron-restricted ablation of NF-kappaB-driven gene expression increases neurodegeneration. This might lead to the path for new treatments of neurodegenerative diseases. PMID- 14561869 TI - An unbiased cDNA library prepared from isolated Aplysia sensory neuron processes is enriched for cytoskeletal and translational mRNAs. AB - Local protein synthesis is required for long-lasting synapse-specific plasticity in cultured Aplysia sensorimotor synapses. To identify synaptically localized mRNAs, we prepared a cDNA library from isolated sensory neurites. By sequence analysis, we estimate that the library contains 263 distinct mRNAs, with 98 of these mRNAs constituting 70% of all clones. The localized transcripts are enriched for mRNAs encoding cytoskeletal elements and components of the translational machinery. In situ hybridization confirms that the mRNAs for at least eight of these transcripts are present in distal neurites. Immunocytochemistry reveals that serotonin regulates the translation of one of the localized mRNAs, that encoding alpha1-tubulin. Our identification of mRNAs encoding cytoskeletal elements suggests that local protein synthesis is required for the growth of new synaptic connections associated with persistent synaptic strengthening. Our finding of mRNAs encoding components of the translational machinery suggests that local protein synthesis serves to increase the translational capacity of synapses. PMID- 14561870 TI - Histone deacetylase inhibition by sodium butyrate chemotherapy ameliorates the neurodegenerative phenotype in Huntington's disease mice. AB - The precise cause of neuronal death in Huntington's disease (HD) is unknown. Although no single specific protein-protein interaction of mutant huntingtin has emerged as the pathologic trigger, transcriptional dysfunction may contribute to the neurodegeneration observed in HD. Pharmacological treatment using the histone deacetylase inhibitor sodium butyrate to modulate transcription significantly extended survival in a dose-dependent manner, improved body weight and motor performance, and delayed the neuropathological sequelae in the R6/2 transgenic mouse model of HD. Sodium butyrate also increased histone and Specificity protein 1 acetylation and protected against 3-nitropropionic acid neurotoxicity. Microarray analysis showed increased expression of alpha- and beta-globins and MAP kinase phosphatase-1 in sodium butyrate-treated R6/2 mice, indicative of improved oxidative phosphorylation and transcriptional regulation. These findings strengthen the hypothesis that transcriptional dysfunction plays a role in the pathogenesis of HD and suggest that therapies aimed at modulating transcription may target early pathological events and provide clinical benefits to HD patients. PMID- 14561871 TI - Functional repair of the corticospinal tract by delayed transplantation of olfactory ensheathing cells in adult rats. AB - Adult rats were trained to use their forepaws to retrieve a piece of food. Destruction of the dorsal corticospinal tract on one side at the level of the first cervical segment abolished the use of the ipsilateral forepaw for retrieval for at least 6 months after operation. Where a variable amount of the corticospinal tract was spared, there was a proportionate persistence of retrieval. In lesioned rats that had shown no retrieval for 8 weeks after operation, a suspension of olfactory ensheathing cells was injected into the lesion site. Starting between 1 and 3 weeks after transplantation, all rats with transplants bridging the lesion site resumed retrieval by the ipsilateral forepaw. Biotin dextran anterograde tracing shows regenerating corticospinal axons crossing the bridge, traveling caudally for approximately 10 mm in the distal part of the corticospinal tract and forming terminal arborizations in the spinal gray matter. Functional recovery can occur when only approximately 1% of the corticospinal tract axons are present. PMID- 14561872 TI - A region on chromosome 15 controls intersession habituation in mice. AB - Habituation to a novel environment, as measured by a change in exploratory activity over time, can be measured both within (intrasession) and across (intersession) sessions. The role of genetics in intrasession habituation has been investigated previously in quantitative trait loci studies, but little attention has been focused on the role of genetics on intersession habituation. We reported recently that inbred strains respond differently in an intersession habituation test. By testing a total of 25 BXD recombinant inbred lines, we were able to map a chromosomal region that strongly influences the way in which mice habituate. This region located on chromosome 15 appears to the major one affecting habituation and accounts for 80% of the genetic variance. We subsequently confirmed this map position by testing (C57BL/6J x DBA/2J) F2 mice. PMID- 14561873 TI - Human hippocampal and parahippocampal activity during visual associative recognition memory for spatial and nonspatial stimulus configurations. AB - Evidence from animal studies points to the importance of the parahippocampal region (PHR) [including entorhinal, perirhinal, and parahippocampal (PHC) cortices] for recognition of visual stimuli. Recent findings in animals suggest that PHR may also be involved in visual associative recognition memory for configurations of stimuli. Thus far, however, such involvement has not been demonstrated in humans. In fact, it has been argued that associative recognition in humans is critically dependent on the hippocampal formation (HF). To better understand the division of function between HF and PHR during recognition memory in humans, we measured the activity of both areas in healthy young adults during an associative recognition memory task using functional magnetic resonance imaging. To more precisely characterize the nature of the associations that might be coded by the HF and PHR during recognition, subjects were required to learn and were later tested for associations based on either the spatial arrangements of two stimuli or the identity of two stimuli (a face and a tool). An area in the PHC was found to be more active for recognized old configurations than new configurations in both the spatial and identity conditions. The HF, on the other hand, was more active for recognition of new configurations than old configurations and also more active in the spatial than the identity condition. These data highlight the involvement of PHR in the long-term coding of associative relationships between stimuli and help to clarify the nature of its functional distinction from the HF. PMID- 14561875 TI - Neurofilament transport in vivo minimally requires hetero-oligomer formation. AB - Neurofilament assembly requires at minimum the polymerization of neurofilament light chain (NF-L) with either neurofilament medium chain (NF-M) or neurofilament heavy chain (NF-H) subunits, but requirements for their axonal transport have long been controversial. Using a gene deletion approach, we generated mice containing only NF-L or NF-M. In vivo pulse radiolabeling analyses in retinal ganglion cell neurons revealed that NF-L alone is incapable of efficient transport, whereas nearly one-half of the normal level of NF-M is transported along optic axons in the absence of the other triplet subunits. Under these conditions, however, NF-M transport is completely abolished by deleting alpha internexin. Our results strongly suggest that efficient neurofilament protein transport in vivo minimally requires hetero-oligomer formation. They also show that NF-M can partner with intermediate filament proteins other than the NF-H and NF-L subunits in neurons to support slow transport and possibly other functions of neuronal intermediate filaments. PMID- 14561874 TI - The bed nucleus is a neuroanatomical substrate for the anorectic effect of corticotropin-releasing factor and for its reversal by nociceptin/orphanin FQ. AB - Nociceptin/orphanin FQ (N/OFQ), the endogenous ligand of the opioid N/OFQ receptor (NOP), possesses marked functional anti-stress and anti-corticotropin releasing factor (CRF) actions. We have shown that intracerebroventricular injection of N/OFQ reverses the hypophagic effect induced by stress or by CRF given intracerebroventricularly. To shed new light on the mechanisms involved in the anti-CRF action of N/OFQ, we investigated the ability of N/OFQ to prevent CRF induced anorexia after microinjection studies into brain areas of potential interest in the control of feeding behavior and coexpressing NOP and CRF receptors. These areas include the bed nucleus of the stria terminalis (BNST), the central amygdala (CeA), the locus ceruleus (LC), the ventromedial hypothalamus (VMH), the paraventricular nucleus (PVN), and the dorsal raphe (DR). The results demonstrated that the anorectic effect of 0.04 nmol of CRF per rat (200 ng per rat) given intracerebroventricularly is reversed by pretreatment with 0.01-0.21 nmol of N/OFQ per rat (25-500 ng per rat) injected into the BNST but not into the CeA, LC, VMH, PVN, or DR. Microinjection of 0.01-0.02 nmol of CRF per site (50-100 ng per site) into the BNST but not into the CeA or the LC induced marked anorexia in food-deprived rats. Pretreatment with 0.01-0.21 nmol of N/OFQ per site (25-500 ng per site) into the BNST also blocked the anorectic action of 0.02 nmol of CRF per site (100 ng per site) given in the same area. Finally, intra-BNST microinjection of 0.01-0.21 nmol of N/OFQ per site (25-500 ng per site) did not modify food intake in either food-sated or food-deprived rats. These data demonstrate that the BNST is involved in the modulation of CRF-induced anorexia, which is prevented by activation of N/OFQ receptors. PMID- 14561876 TI - Complement activation contributes to hypoxic-ischemic brain injury in neonatal rats. AB - Conflicting data have emerged regarding the role of complement activation in the pathophysiology of cerebral ischemia. On the basis of considerable evidence implicating inflammatory mediators in the progression of neonatal brain injury, we evaluated the contribution of complement activation to cerebral hypoxic ischemic (HI) injury in the neonatal rat. To elicit unilateral forebrain HI injury, 7-d-old rats underwent right carotid ligation followed by 1.5-2 hr of exposure to 8% oxygen. Using immunoprecipitation and Western blot assays, we determined that HI induces local complement cascade activation as early as 8 hr post-HI; there was an eightfold increase in the activation fragment inactivated C3b at 16 hr. With immunofluorescence assays and confocal microscopy, both C3 and C9 were localized to injured neurons 16 and 24 hr post-HI. To investigate the contribution of systemic complement to brain injury, we administered the complement-depleting agent cobra venom factor (CVF) 24 hr before HI lesioning and evaluated both acute HI-induced complement deposition and the extent of resulting tissue injury 5 d after lesioning. CVF depleted both systemic and brain C3 by the time of surgery and reduced infarct size. Analysis of lesioned, CVF-treated animals demonstrated minimal neuronal C3 deposition but no reduction in C9 deposition. C3-immunoreactive microglia were identified in injured areas. These results indicate that complement activation contributes to HI injury in neonatal rat brain, systemic administration of CVF does not eliminate complement deposition within injured brain, and microglia may represent an important local source of C3 after acute brain injury. PMID- 14561877 TI - Planar and vertical signals control cellular differentiation and patterning in the mammalian cochlea. AB - The sensory epithelium of the mammalian cochlea is composed of a regular mosaic of sensory hair cells and nonsensory supporting cells. During development, differentiation occurs in a gradient that progresses along the axis of the cochlea from base to apex. To begin to identify some of the factors that regulate this developmental process, the potential roles of planar and vertical signals were examined during early stages of cochlear development. We demonstrate roles for both underlying mesenchymal cells and adjacent epithelial cells in the differentiation and patterning of the sensory epithelium, and in particular in the development of mechanosensory hair cells. As development proceeds, the requirements for both planar and vertical signals decrease, and development of the sensory epithelium becomes essentially independent from these cues. Finally, we demonstrate that the temporal gradient of cellular differentiation is not dependent on planar signals within the developing sensory epithelium. PMID- 14561879 TI - Ribosomal dynamics inferred from variations in experimental measurements. AB - The crystal structures of the ribosome reveal remarkable complexity and provide a starting set of snapshots with which to understand the dynamics of translation. To augment the static crystallographic models with dynamic information present in crosslink, footprint, and cleavage data, we examined 2691 proximity measurements and focused on the subset that was apparently incompatible with >40 published crystal structures. The measurements from this subset generally involve regions of the structure that are functionally conserved and structurally flexible. Local movements in the crystallographic states of the ribosome that would satisfy biochemical proximity measurements show coherent patterns suggesting alternative conformations of the ribosome. Three different types of data obtained for the two subunits display similar "mismatching" patterns, suggesting that the signals are robust and real. In particular, there is an indication of coherent motion in the decoding region within the 30S subunit and central protuberance and surrounding areas of the 50S subunit. Directions of rearrangements fluctuate around the proposed path of tRNA translocation and the plane parallel to the interface of the two subunits. Our results demonstrate that systematic combination and analysis of noisy, apparently incompatible data sources can provide biologically useful signals about structural dynamics. PMID- 14561880 TI - Coincident Hfq binding and RNase E cleavage sites on mRNA and small regulatory RNAs. AB - The Escherichia coli RNA chaperone Hfq was discovered originally as an accessory factor of the phage Qbeta replicase. More recent work suggested a role of Hfq in cellular physiology through its interaction with ompA mRNA and small RNAs (sRNAs), some of which are involved in translational regulation. Despite their stability under certain conditions, E. coli sRNAs contain putative RNase E recognition sites, that is, A/U-rich sequences and adjacent stem-loop structures. We show herein that an RNase E cleavage site coincides with the Hfq-binding site in the 5'-untranslated region of E. coli ompA mRNA as well as with that in the sRNA, DsrA. Likewise, Hfq protects RyhB RNA from in vitro cleavage by RNase E. These in vitro data are supported by the increased abundance of DsrA and RyhB sRNAs in an RNase E mutant strain as well as by their decreased stability in a hfq(-) strain. It is commonly believed that the RNA chaperone Hfq facilitates or promotes the interaction between sRNAs and their mRNA targets. This study reveals another role for Hfq, that is, protection of sRNAs from endonucleolytic attack. PMID- 14561881 TI - Selection of the simplest RNA that binds isoleucine. AB - We have identified the simplest RNA binding site for isoleucine using selection amplification (SELEX), by shrinking the size of the randomized region until affinity selection is extinguished. Such a protocol can be useful because selection does not necessarily make the simplest active motif most prominent, as is often assumed. We find an isoleucine binding site that behaves exactly as predicted for the site that requires fewest nucleotides. This UAUU motif (16 highly conserved positions; 27 total), is also the most abundant site in successful selections on short random tracts. The UAUU site, now isolated independently at least 63 times, is a small asymmetric internal loop. Conserved loop sequences include isoleucine codon and anticodon triplets, whose nucleotides are required for amino acid binding. This reproducible association between isoleucine and its coding sequences supports the idea that the genetic code is, at least in part, a stereochemical residue of the most easily isolated RNA-amino acid binding structures. PMID- 14561882 TI - A template-proximal RNA paired element contributes to Saccharomyces cerevisiae telomerase activity. AB - The ribonucleoprotein complex telomerase is critical for replenishing chromosome end sequence during eukaryotic DNA replication. The template for the addition of telomeric repeats is provided by the RNA component of telomerase. However, in budding yeast, little is known about the structure and function of most of the remainder of the telomerase RNA. Here, we report the identification of a paired element located immediately 5' of the template region in the Saccharomyces cerevisiae telomerase RNA. Mutations disrupting or replacing the helical element showed that this structure, but not its exact nucleotide sequence, is important for telomerase function in vivo and in vitro. Biochemical characterization of a paired element mutant showed that the mutant generated longer products and incorporated noncognate nucleotides. Sequencing of in vivo synthesized telomeres from this mutant showed that DNA synthesis proceeded beyond the normal template. Thus, the S. cerevisiae element resembles a similar element found in Kluyveromyces budding yeasts with respect to a function in template boundary specification. In addition, the in vitro activity of the paired element mutant indicates that the RNA element has additional functions in enzyme processivity and in directing template usage by telomerase. PMID- 14561883 TI - Structural organization of a viral IRES depends on the integrity of the GNRA motif. AB - Little is known about the tertiary structure of internal ribosome entry site (IRES) elements. The central domain of foot-and-mouth disease (FMDV) IRES, named 3 or I, contains a conserved GNRA motif, essential for IRES activity. We have combined functional analysis with RNA probing to define its structural organization. We have found that a UNCG motif does not functionally substitute the GNRA motif; moreover, binding of synthetic GNRA stem-loops to domain 3 was significantly reduced in RNAs bearing UCCG or GUAG substitutions. The apical region of domain 3 consists of a four-way junction where residues of the GNRA tetraloop are responsible for the organization of the adjacent stem-loops, as deduced from ribonucleases and dimethyl sulfate accessibility. A single A-to-G substitution in the fourth position of this motif led to a strong RNA reorganization, affecting several nucleotides away in the secondary structure of domain 3. The study of mutants bearing UNCG or GUAG tetraloops revealed lack of protection to chemical attack in native RNA at specific nucleotides relative to the parental GUAA, suggesting that the GNRA motif dictates the organization and stability of domain 3. This effect is likely mediated by the interaction with distant residues. Therefore, the GNRA motif plays a crucial role in the organization of IRES structure with important consequences on activity. PMID- 14561884 TI - Structures of deacylated tRNA mimics bound to the E site of the large ribosomal subunit. AB - During translation, tRNAs cycle through three binding sites on the ribosome: the A, the P, and the E sites. We have determined the structures of complexes between the Haloarcula marismortui large ribosomal subunit and two different E site substrates: a deacylated tRNA acceptor stem minihelix and a CCA-acceptor end. Both of these tRNA mimics contain analogs of adenosine 76, the component responsible for a large proportion of E site binding affinity. They bind in the center of the loop-extension of protein L44e, and make specific contacts with both L44e and 23S rRNA including bases that are conserved in all three kingdoms of life. These contacts are consistent with the footprinting, protection, and cross-linking data that have identified the E site biochemically. These structures explain the specificity of the E site for deacylated tRNAs, as it is too small to accommodate any relevant aminoacyl-tRNA. The orientation of the minihelix suggests that it may mimic the P/E hybrid state. It appears that the E site on the 50S subunit was formed by only RNA in the last common ancestor of the three kingdoms, since the proteins at the E sites of H. marismortui and Deinucoccus radiodurans large subunits are not homologous. PMID- 14561885 TI - Visualization of membrane RNAs. AB - Using fluorescence microscopy, we show that previously isolated membrane-binding RNAs coat artificial phospholipid membranes relatively uniformly, except for a frequent tendency to concentrate at bends, membrane junctions, and other unusual sites. Membrane RNAs can also be visualized as single molecules or isolated complexes by atomic force microscopy (AFM) of free RNAs on mica. Finally, RNAs can be seen within membranes by AFM of RNA-liposomes immobilized on hydrophobic mica surfaces. Monomer RNAs appear globular, as expected for small RNAs. When mixed under conditions in which RNAs bind bilayers, RNA 9 and RNA 10 combine to yield about 80% of RNAs as mainly linear oligomers of approximately 2-8 molecules. Once inserted in membranes, the RNAs oligomerize further, yielding larger, irregular ropelike structures that prefer the edges of altered lipid patches. These properties can be interpreted in terms of RNA-RNA loop interactions, and the RNA effects on membranes can be explained in terms of an RNA preference for irregular lipid conformations. The RNA-bilayer system poses new opportunities for combining the properties of membranes and RNA in contemporary cells, and also in the ribocytes of an RNA world. PMID- 14561886 TI - The roles of endonucleolytic cleavage and exonucleolytic digestion in the 5'-end processing of S. cerevisiae box C/D snoRNAs. AB - Small nucleolar RNAs (snoRNAs) play important roles in ribosomal RNA metabolism. In Saccharomyces cerevisiae, box C/D snoRNAs are synthesized from excised introns, polycistronic precursors, or independent transcription units. Previous studies have shown that only a few independently transcribed box C/D snoRNAs are processed at their 5' end. Here we describe 12 additional independently transcribed box C/D snoRNAs that undergo 5'-end processing. 5' Extensions found in the precursors of these snoRNAs contain cleavage sites for Rnt1p, the S. cerevisiae homolog of RNase III, and unprocessed precursors accumulate in vivo in the absence of Rnt1p. Rnt1p cleavage products were identified in vivo when the 5' --> 3' exonucleases Xrn1p and Rat1p are inactivated (xrn1delta rat1-1) and in vitro using model RNA substrates and recombinant Rnt1p. Some of these snoRNAs show increased levels of unprocessed precursors when the rnt1Delta deletion is combined to the xrn1delta rat1-1 mutation, suggesting that these exonucleases participate in the 5' processing or the degradation of the snoRNA precursors. Unprocessed precursors are not significantly destabilized in the absence of the trimethylguanosine capping enzyme Tgs1p, suggesting that a 5' monomethyl cap is sufficient to ensure stabilization of these precursors. These results demonstrate that the majority of independently transcribed box C/D snoRNAs from the yeast genome undergo 5'-end processing and that the Rnt1p endonuclease and the Xrn1p and Rat1p 5' --> 3'exonucleases have partially redundant functions in the 5'-end processing of these snoRNAs. PMID- 14561887 TI - The Saccharomyces cerevisiae U2 snRNA:pseudouridine-synthase Pus7p is a novel multisite-multisubstrate RNA:Psi-synthase also acting on tRNAs. AB - The Saccharomyces cerevisiae Pus7 protein was recently characterized as a novel RNA:pseudouridine (Psi)-synthase acting at position 35 in U2 snRNA. However, U2 snRNA was the only potential substrate tested for this enzyme. In this work, we demonstrated that although Pus7p is responsible for the formation of only one of the six Psi residues present in yeast UsnRNAs, it catalyzes U to Psi conversion at position 13 in cytoplasmic tRNAs and at position 35 in pre-tRNA(Tyr). Sites of RNA modification by Pus7p were identified by analysis of the in vivo RNA modification defects resulting from the absence of active Pus7p production and by in vitro tests using extracts from WT and genetically modified yeast cells. For demonstration of the direct implication of Pus7p in RNA modification, the activity of the WT and mutated Pus7p recombinant proteins was tested on in vitro produced tRNA and pre-tRNA transcripts. Mutation of an aspartic acid residue (D256) that is conserved in all Pus7 homologs abolishes the enzymatic activity both in vivo and in vitro. This suggests the direct involvement of D256 in catalysis. Target sites of Pus7p in RNAs share a common sequence Pu(G/C)UNPsiAPu (Pu = purine, N = any nucleotide), which is expected to be important for substrate recognition. Modification of tRNAs by Pus7p explains the presence of Pus7p homologs in archaea and some bacteria species, which do not have U2 snRNA, and in vertebrates, where Psi34 (equivalent to Psi35 in yeast) formation in U2 snRNA is an H/ACA snoRNA guided process. Our results increase the number of known RNA modification enzymes acting on different types of cellular RNAs. PMID- 14561888 TI - RNA-protein interactions promote asymmetric sorting of the ASH1 mRNA ribonucleoprotein complex. AB - In Saccharomyces cerevisiae, ASH1 mRNA is localized to the tip of daughter cells during anaphase of the cell cycle. ASH1 mRNA localization is dependent on four cis-acting localization elements as well as Myo4p, She2p, and She3p. Myo4p, She2p, and She3p are hypothesized to form a heterotrimeric protein complex that directly transports ASH1 mRNA to daughter cells. She2p is an RNA-binding protein that directly interacts with ASH1 cis-acting localization elements and associates with She3p. Here we report the identification of seven She2p mutants-N36S, R43A, R44A, R52A, R52K, R63A, and R63K-that result in the delocalization of ASH1 mRNA. These mutants are defective for RNA-binding activity but retain the ability to interact with She3p, indicating that a functional She2p RNA-binding domain is not a prerequisite for association with She3p. Furthermore, the nuclear/cytoplasmic distribution for the N36S and R63K She2p mutants is not altered, indicating that nuclear/cytoplasmic trafficking of She2p is independent of RNA-binding activity. Using the N36S and R63K She2p mutants, we observed that in the absence of She2p RNA-binding activity, neither Myo4p nor She3p is asymmetrically sorted to daughter cells. However, in the absence of She2p, Myo4p and She3p can be asymmetrically segregated to daughter cells by artificially tethering mRNA to She3p, implying that the transport and/or anchoring of the Myo4p/She3p complex is dependent on the presence of associated mRNA. PMID- 14561889 TI - Association of polyadenylation cleavage factor I with U1 snRNP. AB - Splicing and polyadenylation factors interact for the control of polyadenylation and the coupling of splicing and polyadenylation. We document an interaction between the U1 snRNP and mammalian polyadenylation cleavage factor I (CF Im), one of several polyadenylation factors needed for the cleavage of the pre-mRNA at the polyadenylation site. Sucrose density gradient centrifugation demonstrated that CF Im separated into two fractions, a light fraction which contained the known CF Im subunits (72, 68, 59, and 25 kD), and a heavy fraction, rich in snRNPs, which contained predominately the 68- and 25-kD CF Im subunits. Using specific antibodies we found that the heavy fraction contains U1 snRNP/CF Im coprecipitable complexes. These complexes were insensitive to RNase treatment, suggesting that the coprecipitation is not due to RNA tethering. In vitro binding experiments show that both the 68- and 25-kD subunits bind to and comigrate with U1 snRNP. In addition, the 25-kD CF Im subunit binds specifically to the 70K protein of U1 snRNP (U1 70K). This binding may account for the CF Im/U1 snRNP interaction. During these studies we found that mAb 2.73 (mAb 2.73), an established U1 70K antibody, efficiently precipitates the bulk of the CF Im from cellular extracts. Because mAb 2.73 has been used in a number of previous studies related to the U1 snRNP and the U1 70K protein, the precipitation of CF Im must be considered in evaluating past and future data based on the use of mAb 2.73. PMID- 14561890 TI - Protein ligands mediate the CRM1-dependent export of HuR in response to heat shock. PMID- 14561891 TI - Differentiation of human embryonic stem cells on three-dimensional polymer scaffolds. AB - Human embryonic stem (hES) cells hold promise as an unlimited source of cells for transplantation therapies. However, control of their proliferation and differentiation into complex, viable 3D tissues is challenging. Here we examine the use of biodegradable polymer scaffolds for promoting hES cell growth and differentiation and formation of 3D structures. We show that complex structures with features of various committed embryonic tissues can be generated, in vitro, by using early differentiating hES cells and further inducing their differentiation in a supportive 3D environment such as poly(lactic-co-glycolic acid)/poly(L-lactic acid) polymer scaffolds. We found that hES cell differentiation and organization can be influenced by the scaffold and directed by growth factors such as retinoic acid, transforming growth factor beta, activin A, or insulin-like growth factor. These growth factors induced differentiation into 3D structures with characteristics of developing neural tissues, cartilage, or liver, respectively. In addition, formation of a 3D vessel-like network was observed. When transplanted into severe combined immunodeficient mice, the constructs continue to express specific human proteins in defined differentiated structures and appear to recruit and anastamose with the host vasculature. This approach provides a unique culture system for addressing questions in cell and developmental biology, and provides a potential mechanism for creating viable human tissue structures for therapeutic applications. PMID- 14561892 TI - A modulatory role for facial expressions in prosopagnosia. AB - Brain-damaged patients experience difficulties in recognizing a face (prosopagnosics), but they can still recognize its expression. The dissociation between these two face-related skills has served as a keystone of models of face processing. We now report that the presence of a facial expression can influence face identification. For normal viewers, the presence of a facial expression influences performance negatively, whereas for prosopagnosic patients, it improves performance dramatically. Accordingly, although prosopagnosic patients show a failure to process the facial configuration in the interest of face identification, that ability returns when the face shows an emotional expression. Accompanying brain-imaging results indicate activation in brain areas (amygdala, superior temporal sulcus, parietal cortex) outside the occipitotemporal areas normally activated for face identification and lesioned in these patients. This finding suggests a modulatory role of these areas in face identification that is independent of occipitotemporal face areas. PMID- 14561893 TI - Early phagosomes in dendritic cells form a cellular compartment sufficient for cross presentation of exogenous antigens. AB - Conventionally, MHC class I-restricted antigen (Ag) processing requires the action of the multimolecular peptide-loading complex within the endoplasmic reticulum (ER). Here we show that early phagosomes from human dendritic cells (DCs) contain the peptide-loading complex, incorporating MHC class I, beta2 microglobulin, transporter associated with Ag processing (TAP), calreticulin, tapasin, and ERp57. Antigenic peptides could be translocated into purified phagosomes by TAP and loaded onto cognate class I molecules, inducing their specific dissociation from the loading complex. Endoglycosidase H-sensitive class I molecules were detected at the DC cell surface, suggesting that these molecules traffic there directly from phagosomes. Macropinocytosis also allowed internalized soluble Ags access to an ER-like compartment containing the class I loading complex. Blockade of TAP by endocytosis of a soluble derivative of human cytomegalovirus protein US6 confirmed that, although retrotranslocation into the cytosol is critical for processing, efficient association of class I molecules with peptides derived from exogenous Ags occurs within a compartment directly accessible to internalized proteins. Together, this evidence suggests that early phagosomes and pinosomes facilitate cross presentation of exogenous Ags by DCs. PMID- 14561894 TI - Alu elements and hominid phylogenetics. AB - Alu elements have inserted in primate genomes throughout the evolution of the order. One particular Alu lineage (Ye) began amplifying relatively early in hominid evolution and continued propagating at a low level as many of its members are found in a variety of hominid genomes. This study represents the first conclusive application of short interspersed elements, which are considered nearly homoplasy-free, to elucidate the phylogeny of hominids. Phylogenetic analysis of Alu Ye5 elements and elements from several other subfamilies reveals high levels of support for monophyly of Hominidae, tribe Hominini and subtribe Hominina. Here we present the strongest evidence reported to date for a sister relationship between humans and chimpanzees while clearly distinguishing the chimpanzee and human lineages. PMID- 14561895 TI - Interaction between the Alzheimer's survival peptide humanin and insulin-like growth factor-binding protein 3 regulates cell survival and apoptosis. AB - Insulin-like growth factor-binding protein-3 (IGFBP-3) regulates IGF bioactivity and also independently modulates cell growth and survival. By using a yeast two hybrid screen to identify IGFBP-3-interacting proteins, we cloned humanin (HN) as an IGFBP-3-binding partner. HN is a 24-aa peptide that has been shown to specifically inhibit neuronal cell death induced by familial Alzheimer's disease mutant genes and amyloid-beta (Abeta). The physical interaction of HN with IGFBP 3 was determined to be of high affinity and specificity and was confirmed by yeast mating, displaceable pull-down experiments with (His)-6-tagged HN, and ligand blot experiments. Co-immunoprecipitation of IGFBP-3 and HN from mouse testes confirmed the interaction in vivo. In cross-linking experiments, HN bound IGFBP-3 but did not compete with IGF-I-IGFBP-3 binding; competitive ligand dot blot experiments revealed the 18-aa heparin-binding domain of IGFBP-3 as the binding site for HN. Alanine scanning determined that F6A-HN mutant does not bind IGFBP-3. HN but not F6A-HN inhibited IGFBP-3-induced apoptosis in human glioblastoma-A172. In contrast, HN did not suppress IGFBP-3 response in SH-SY5Y neuroblastoma and mouse cortical primary neurons. In primary neurons, IGFBP-3 markedly potentiated HN rescue ability from Abeta1-43 toxicity. In summary, we have identified an interaction between the survival peptide HN and IGFBP-3 that is pleiotrophic in nature and is capable of both synergistic and antagonistic interaction. This interaction may prove to be important in neurological disease processes and could provide important targets for drug development. PMID- 14561896 TI - Thrombospondin 1, a mediator of the antiangiogenic effects of low-dose metronomic chemotherapy. AB - Chemotherapeutic drugs chronically administered to tumor-bearing mice, using a frequent schedule at doses substantially lower than the maximum tolerated dose (MTD) (i.e., metronomic dosing), can cause sustained and potent antiangiogenic effects by targeting the endothelial cells of newly growing tumor blood vessels. These effects appear to occur in the absence of an increase in the severity of side effects caused by destruction of other cell types normally sensitive to MTD chemotherapy, suggesting a marked and selective sensitivity of activated endothelial cells, the basis of which is unknown. Here we report that protracted exposure of endothelial cells in vitro to low concentrations of several different anticancer agents, including microtubule inhibitors and an alkylating agent, caused marked induction of gene and protein expression of TSP-1, a potent and endothelial-specific inhibitor of angiogenesis. Increases in circulating TSP-1 were also detected in the plasma of human tumor-bearing severe combined immunodeficient mice treated with metronomic low-dose cyclophosphamide. Most importantly, the antiangiogenic and antitumor effects of low-dose continuous cyclophosphamide were lost in TSP-1-null C57BL/6 mice, whereas, in contrast, these effects were retained by using a MTD schedule of the same drug. Taken together, the results implicate TSP-1 as a secondary mediator of the antiangiogenic effects of at least some low-dose metronomic chemotherapy regimens. PMID- 14561897 TI - Influenza virus hemagglutinin concentrates in lipid raft microdomains for efficient viral fusion. AB - Lipid raft microdomains are enriched in sphingomyelin and cholesterol and function as platforms for signal transduction and as the site of budding of several enveloped viruses, including influenza virus. The influenza virus hemagglutinin (HA) glycoprotein, which mediates both viral-cell attachment and membrane fusion, associates intrinsically with lipid rafts. Residues in the HA transmembrane (TM) domain are important for raft association as sequence substitutions in the HA TM domain ablate HA association with rafts (nonraft HA). Cells expressing either WT or nonraft HA cause complete fusion (lipid mixing and content mixing) over widely varying HA expression levels. However, the number of fusion events measured for nonraft HA mutant protein at all HA surface densities was reduced to approximately 55% of the events for WT HA protein. Mutant influenza viruses were generated that contain the nonraft HA TM domain alterations. Electron microscopy experiments showed that WT HA was distributed at the cell surface in clusters of 200-280 nm in diameter, whereas nonraft HA was distributed mostly randomly at the plasma membrane. Nonraft HA virus showed reduced budding, contained reduced amounts of HA protein, was greatly reduced in infectivity, and exhibited decreased virus-membrane fusion activity. Cholesterol depletion of virus did not affect the ability of virions to cause either virus cell lipid mixing or virus-mediated hemolysis, a surrogate for content mixing. Taken together, the data suggest that HA clusters in rafts to provide a sufficient concentration of HA in budding virus to mediate efficient virus-cell fusion. PMID- 14561899 TI - Haemorheological pattern in young adults with acute myocardial infarction. AB - Acute myocardial infarction (AMI) in young people has recently received great attention. Its main characteristics include a typical risk factor profile and a good prognosis. We studied the haemorheological pattern (whole blood viscosity at high and low shear rates, plasma and serum viscosity, whole-blood filtration and erythrocyte aggregation) in 64 subjects, aged <46 years, with recent AMI. We observed marked alterations in blood, plasma and serum viscosity and whole-blood filtration. Subdividing the AMI subjects in accordance with the number of cardiovascular risk factors or the extent of coronary lesions, we did not observe any significative influence of these aspects on the haemorheological determinants, with the exception of low shear rate blood viscosity. The latter was in fact higher than in control subjects only in AMI subgroups with respectively more risk factors and more stenosed coronary vessels. Thirty-three AMI subjects were re-examined after three months and showed, on this occasion, a haemorheological pattern not significantly different from that of the first evaluation. These results demonstrate that in young subjects with AMI there is a hyperviscosity syndrome that persists during subsequent months, despite a good clinical course. Haemorheological impairment may unfavourably influence the long term prognosis of AMI in young subjects and therefore, in our opinion, management should take into account the monitoring of the haemorheological pattern. PMID- 14561898 TI - Impaired plasma viscosity via increased cholesterol levels in peripheral occlusive arterial disease [correction of disase]. AB - The aim of this study was to investigate the relationship between plasma viscosity and lipoprotein and apolipoprotein pattern in normo- and hypercholesterolemic patients with peripheral occlusive arterial disease (POAD). 40 patients with POAD have been selected (8 females and 32 males, mean age: 54+/ 3.2 years) with clinically evident superficial femoral occlusive artery disease. They were separated into two groups as normocholesterolemic (plasma total cholesterol <200 mg/dl) and hypercholesterolemic (plasma total cholesterol >200 mg/dl). Plasma total cholesterol, high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), triglycerides, total protein, and albumin levels were determined by enzymatic methods using commercial kits. Levels of apolipoprotein AI (apo AI), and apolipoprotein B (apo B) were measured using a immunoturbidometric method. Plasma viscosity (PV) was measured by capillary viscometer. Classifying the patients with PAOD according to the cholesterol levels; hypercholesterolemic (mean total-cholesterol: 227.90+/-26.97 mg/dl) patients had significantly higher LDL-C, PV and triglyceride levels compared with nornocholesterolemic patients (p<0.001, p<0.001, p<0.001, respectively). HDL-C and apo B were significantly lower in hypercholesterolemic patients than in normocholesterolemic patients (p<0.001, p<0.001, respectively). PV was positively correlated with total cholesterol (r=0.485, p<0.05), atherogenic index (r=0.624, p<0.01), total-C/HDL-C ratio (r=0.624, p<0.05), and LDL-C/HDL-C ratio (r=0.707, p<0.001) in hypercholesterolemic patients with POAD. PV was higher in hypercholesterolemic patients with POAD than in normocholesterolemic patients with POAD. We suggest that POAD patients should be regarded as a heterogenous group with lipid and lipoprotein parameters in order to assess the microcirculation in the affected limb. In case of dyslipidemia in POAD patients an elevated plasma viscosity should be considered as coexisting risk factor. PMID- 14561900 TI - Relationships between von Willebrand factor and hemorheology in sportsmen. AB - It is well established that exercise performance in athletes is related to improved blood fluidity. However, training effects on functional state of endothelium and relations of endothelial cell functions with hemorheology are poorly known. Circulating levels of von Willebrand factor may serve as a good marker of endothelial cell functions, its activation, and damage. Relationships between von Willebrand factor antigen (vWf) and blood rheology in 30 endurance sportsmen were investigated. Athletes were divided according to vWf into tertile groups. Compared to 16 controls, all subgroups of sportsmen had a lower erythrocyte rigidity index Tk (p<0.05-0.01), explained by lower values of mean corpuscular hemoglobin concentration (MCHC) (p<0.05-0.01), and a higher W170. The lower tertile group (<0.35 U/ml) had a lower blood viscosity (p<0.05), explained by a lower Tk, and a lower erythrocyte aggregability index (microscopy of diluted blood samples) (p<0.05). The upper tertile group of athletes demonstrated higher levels of plasma viscosity, explained by higher total globulins (p<0.01), and higher vWf levels (p<0.01) than controls, and a lower W170 compared to athletes from the lower tertile subgroup. In the entire group of athletes, log (vWf) was positively correlated to plasma viscosity (r=0.478, p=0.007), total serum globulins (r=0.430, p=0.018), erythrocyte aggregability index (r=0.427, p=0.019), and negatively to log (W170) (r=-0.449, p=0.013). Multiple stepwise regression analysis revealed that plasma viscosity was the primary correlate of vWf. These data shown that the higher von Willebrand level reflecting strong activation of endothelial cells in a part of athletes was closely correlated with increased plasma viscosity levels. We suggest that these hemorheological and endothelial disturbances in these athletes might be a result of exercise overloads. PMID- 14561901 TI - Reference range of hematocrit in the elderly with respect to altitude. AB - The purpose of this paper is to provide a scientific basis for a unified standard of the reference value of old people's hematocrit in China. The reference values of healthy old people's hematocrit have been collected according to the Wintrobe methods; the relationship between the reference values of old people's hematocrit and altitude has been tested in this paper. It has been found that the reference value of old people's hematocrit increases when the altitude gradually increases, and such relationship is quite significant. The method of mathematical univariate regression analysis is used to deduce two regression equations: Y_1=44.1+0.00351x+/-4.5, Y_2=39.5+0.00298x+/-4.1. If the altitude value of a particular area of China is known, the reference value of old people's hematocrit there can be calculated by means of the regression equations. Furthermore, depending on the altitude, China can be divided into three districts: Qingzang District, Central District and Eastern District. PMID- 14561902 TI - Influence of propofol on erythrocyte morphology, blood viscosity and platelet function. AB - The intravenous anaesthetic propofol has been associated with cardiovascular side effects. We therefore studied its influence on blood viscosity, erythrocytes and platelet aggregation. Blood from healthy volunteers was incubated with propofol concentrations of 0, 5, 10, 20, 40, 80, 200, and 500 microg/ml plasma. Whole blood viscosity (shear rates 94.5 and 0.1 s(-1)), plasma viscosity, erythrocyte morphology and platelet aggregation (PFA-100 system) were determined. These parameters were also measured in vivo in 9 patients prior to anaesthesia, after induction, before the end and 1 h after the end of propofol anaesthesia. Propofol induced a slight, dose-dependent echinocytic shape transformation of erythrocytes in vitro, indicating a preferential intercalation of the drug in the outer hemileaflet of the membrane. Neither whole blood nor plasma viscosity were affected in vitro. In vivo, no change in erythrocytes shape was seen, but plasma and whole blood viscosity at high shear rate (94.5 s(-1)) were decreased at the end of anaesthesia, which may be due to some extent to plasma dilution. Platelet aggregation with epinephrine was decreased both in vitro and in vivo. We conclude that propofol interacts with the erythrocyte membrane without affecting blood and plasma viscosity and decreases platelet aggregation, which may have clinical implications. PMID- 14561904 TI - Influence of radiographic contrast media on myocardial tissue oxygen tension: NaCl-controlled, randomised, comparative study of iohexol versus iopromide in an animal model. AB - It was tested whether a bolus injection of 10 ml radiographic contrast medium (iopromide vs. iohexol), compared to a 10 ml NaCl bolus and administered into the left anterior descending artery (LAD) of farm pigs, influenced the tissue pO2 in the territory of this artery. The radiographic contrast media and the NaCl bolus were given in randomised order. The mean pO2 LAD fell from initially 40.3+/-10.9 mmHg to a minimal value of 22.5+/-8.9 mmHg 241+/-44 sec after injecting the iopromide bolus, with this result representing a mean decrease of 44.2% (p=0.0003). The initial pO2 (baseline) was reached again after approximately 10 minutes. The mean pO2 LAD fell from the initial value of 34.5+/-14.6 mmHg to a minimal value of 29.4+/-13.9 mmHg 171.7+/-11.9 sec after injection of the iohexol bolus, with this result representing a mean decrease of 14.8% (p=0.0003). The baseline pO2 was reached again after approx. 5 minutes. The drop in the pO2 after iopromide administration was significantly larger than that after iohexol (p=0.0001), and also the time after which the baseline pO2 is reached again was considerably shorter for iohexol (p=0.001). The two radiographic contrast media did not influence the tissue pO2 in either the territory of the right coronary artery or in skeletal muscle. Injection of a NaCl bolus into the LAD influenced neither the tissue pO2 of the territory of the LAD nor that of the RCA or of the skeletal muscle. The tissue temperature, heart rate and the systolic and diastolic blood pressure were not affected during the three injections. Injection of radiographic contrast media into a coronary artery can lead to a distinct, local microcirculatory impairment in the myocardial territory supplied by this artery. In this case, the extent of the microcirculation impairment seems to depend not only on the viscosity of the contrast media but rather also on its chemotoxicity. PMID- 14561903 TI - Effects of choline on hemorheological properties and NO metabolism of human erythrocytes. AB - PURPOSE OF THE STUDY: To determine the effects of choline on red blood cell membrane properties and NO metabolism. MATERIAL AND METHODS: Aliquots of venous blood from eleven healthy subjects were incubated in vitro with choline concentrations 10(-10) to 10(-3) M. The following parameters were determined: erythrocyte deformability, aggregation and membrane lipid fluidity, plasma K+, Na+, Ca2+, total blood haemoglobin and methemoglobin concentrations. Additionally, plasma and intra-erythrocyte nitrites concentrations were measured. RESULTS: Choline increases erythrocyte deformability at lower shear stresses, decreases erythrocyte aggregation, increases membrane lipid fluidity, and decreases of Na+ plasma concentrations. We also find an increase of nitrites concentration both in the plasma and in the intra-erythrocyte compartment. CONCLUSION: Choline induces changes on erythrocyte membrane properties, Na+ plasma concentration, and NO metabolites concentrations. PMID- 14561905 TI - Helical CT pulmonary angiography predictors of in-hospital morbidity and mortality in patients with acute pulmonary embolism. AB - PURPOSE: To determine if CT variables predict in-hospital morbidity and mortality in patients with pulmonary embolism (PE). MATERIALS AND METHODS: CT scans and charts of 173 patients with CT scans positive for PE were reviewed. CT scans were reviewed for leftward ventricular septal bowing, increased right ventricle (RV) to left ventricle (LV) diameter ratio, clot burden, increased pulmonary artery to aorta diameter ratio, and oligemia. Charts were reviewed for severe morbidity and mortality outcomes: death from pulmonary emboli or any cause, and cardiac arrest. Charts were also reviewed for milder morbidity outcomes: intubation, vasopressor use, or admission to an intensive care unit (ICU) and for multiple comorbidities. RESULTS: No CT predictor was significantly associated with severe morbidity or mortality outcomes. Ventricular septal bowing and increased RV/LV diameter ratio were both associated with subsequent admission to an ICU (P = 0.004 and P = 0.025, respectively). Oligemia (either lung) was associated with subsequent intubation; right lung oligemia was associated with the subsequent use of vasopressors. After controlling for history of congestive heart failure, ischemic heart disease, and pulmonary disease, both septal bowing and an increased RV/LV diameter ratio remained associated with admission to an ICU. CONCLUSION: No CT variables predicted severe in-hospital morbidity and mortality (death from pulmonary embolism, death from any cause, or cardiac arrest) in patients with PE. However, ventricular septal bowing and increased RV/LV diameter ratio were both strongly predictive of less severe morbidity, namely, subsequent ICU admission, and oligemia was associated with subsequent intubation and vasopressor use. PMID- 14561906 TI - Radiological and clinical findings in acute and chronic exogenous lipoid pneumonia. AB - PURPOSE: To describe the radiologic and clinical findings in a group of patients with exogenous lipoid pneumonia focusing on features that differentiate the acute and chronic presentations. MATERIALS AND METHODS: We retrospectively identified 15 patients from a single institution with exogenous lipoid pneumonia 1992-2001. Inclusion criteria were: imaging with chest CT and either CT features diagnostic of lipoid pneumonia or intrapulmonary lipids on pathologic examination. Each chart was reviewed for the clinical presentation, history of ingestion/exposure, predisposing factors, treatment and response to therapy. Initial (n = 13) and follow-up (n = 11) chest radiographs were reviewed for the patterns and distribution of lung parenchymal abnormalities and pleural effusion. Initial (n = 15) and follow-up (n = 7) CT scans were reviewed for lung parenchymal abnormalities (consolidation, ground glass opacities, linear/nodular opacities, masses, and crazy paving), presence or absence of fat attenuation, and pleural effusion. The groups were compared using the Fischer exact test. RESULTS: Nine patients had acute lipoid pneumonia, 7 males and 2 females with a mean age of 45 (range 4.5-81) years. Six patients had chronic lipoid pneumonia, 4 men and 2 women with a mean age of 63 (range 37-83) years. 78% (7/9) of patients with acute and all of the patients with chronic presentations had a known ingestion and/or a predisposing condition associated with lipoid pneumonia. On chest radiographs, consolidation and lower lobe involvement were present in the majority of patients with acute and none of the patients with chronic presentations. On CT, consolidation and fat attenuation were present in the majority of patients with each presentation. However, masses were present in 67% (4/6) of patients with chronic and none of the patients with acute presentation (P < 0.05). 86% (6/7) of patients with acute presentation had improvement on follow-up chest radiograph in contrast to none of the patients with chronic presentation (P < 0.05). 75% (3/4) of patients with acute lipoid pneumonia had improvement on follow-up CT in contrast with 67% (2/3) of patients with chronic lipoid pneumonia who had progression on follow-up CT. CONCLUSIONS: The imaging features of acute and chronic lipoid pneumonia overlap with consolidation and lower lobe involvement present in both groups. However, only the patients with acute lipoid pneumonia had pleural effusions and improvement on follow-up. Only the patients with chronic lipoid pneumonia had pulmonary masses. PMID- 14561907 TI - The rates and CT patterns of locoregional recurrence after resection surgery of lung cancer: correlation with histopathology and tumor staging. AB - The purpose of our study was to assess the rates and CT patterns of locoregional recurrence after resection surgery of lung cancers according to histopathology and tumor staging. Three hundred and seventy nine patients who underwent lung resection surgery due to lung cancer in a recent 6 year period were followed up with CT (at 3, 6, 12, 18, 24 months, and then annually after surgery) for evaluation of locoregional tumor recurrence (analysis of hilar or mediastinal lymph nodes and surgical margin including bronchial stump, pleura, and chest wall). The recurrence rates and CT patterns were compared in terms of underlying histopathology and tumor staging. Of 379 patients, 75 (20%) patients had locoregional recurrences. The recurrence rates were higher in squamous cell carcinoma (39/190, 21%) than adenocarcinoma (24/140, 17%) (P = 0.012). The patterns of recurrence in 75 patients were hilar-mediastinal lymph node enlargement (n = 39, 52%), ipsilateral pleural lesion (n = 24, 32%), chest wall lesion (n = 13, 17%), bronchial stump lesion (n = 8, 11%), and bronchial extension including the trachea (n = 3, 4%). Bronchial stump recurrence was seen only in squamous cell carcinoma. Bronchioloalveolar carcinoma did not show any evidence of locoregional recurrence. Pleural (P = 0.0016) and mediastinal nodal (P = 0.001) recurrence, respectively, were more common in N2 than N0 cancers. Chest wall recurrence rates were higher with higher T staging (P < 0.001). The locoregional recurrence of lung cancer occurs in about one fifth of patients who undergo curative resection and is more common in squamous cell carcinoma than in adenocarcinoma. Recurrent patterns are diverse and different according to histopathologic type and pathologic staging of lung cancer. PMID- 14561909 TI - Fractal analysis for quantitative evaluation of diffuse lung abnormalities on chest radiographs: use of sub-ROIs. AB - We evaluated the usefulness of fractal analysis for quantitative evaluation of diffuse lung abnormalities seen on chest radiographs. Regions of interest (ROIs) with 296 x 296 matrix size were selected in the right upper and lower lung zones of digitized chest radiographs. We selected ROIs of 50 mild and 50 severe diffuse lung abnormalities and 50 ROIs of normal lungs. The fractal dimension (FD) for each ROI was defined as the mean value of those obtained from sub-ROIs in each ROI. In the sub-ROIs with 256 x 256, 128 x 128, and 64 x 64 matrix size, the mean values of FDs obtained from the patients with diffuse lung abnormalities were significantly higher than those obtained from the patients with normal lungs (P < 0.001, respectively). The performance of FDs obtained from the sub-ROIs with 256 x 256 matrix size was equal to that obtained from the sub-ROIs with 128 x 128 matrix size (P = 0.980). However, the performance of FDs obtained from the sub ROIs with 128 x 128 matrix size was superior to that obtained from the sub-ROIs with 64 x 64 matrix size (P = 0.0001). Fractal analysis is useful for quantitative evaluation of diffuse lung abnormalities seen on chest radiographs. From our results, the sub-ROIs whose matrix sizes were greater than or equal to 128 x 128 were suggested to be suitable for quantitative evaluation of diffuse lung abnormalities seen on chest radiographs. PMID- 14561908 TI - Methotrexate-induced pulmonary injury: serial CT findings. AB - We describe serial computed tomographic (CT) findings of methotrexate (MTX) induced pulmonary injury. MATERIALS AND METHODS: The cases of 8 patients (3 men and 5 women; mean age 58.6 years, range 16 to 75 years) of clinically diagnosed MTX-induced pulmonary injury were reviewed. Six patients had rheumatoid arthritis, 1 had lupus erythematosus profundus, and 1 had juvenile rheumatoid arthritis. CT findings on admission and at follow-up were evaluated. RESULTS: The most common CT features were diffuse and patchy bilateral ground-glass opacity with (n = 3) or without reticulation (n = 4) and consolidation (n = 1). These opacities showed no predilection for any particular lung zone in 6 patients but did show dependent predilection in 1 patient and upper lobe predilection in 1. Diffuse centrilobular ill-defined nodules were noted in 1 patient, which disappeared on follow-up. During the average post-treatment follow-up period of 31.0 days (range 3 to 76 days), the opacities quickly improved after treatment in 6 patients; however, in 2 patients with pre-existing interstitial pneumonitis the opacities were refractory. CONCLUSION: CT features of MTX-induced pulmonary injury were variable and included diffuse parenchymal opacification, reticular opacities, and centrilobular nodules. These opacities usually responded quickly to treatment; however, those patients with lung fibrosis at presentation may have worse prognosis. PMID- 14561910 TI - Utility of simple radiation dose measurements in the evaluation of different CT scanners used for high-resolution CT. AB - In recent years radiation risk from CT scanning has become an important area of investigation. Many authors have suggested that radiation dose can be decreased without loss of diagnostic information. This dose reduction has primarily been achieved through a decrease in tube current. An area that has received little attention has been variations in dose from different CT scanners. To evaluate this aspect of radiation exposure, we measured the radiation dose of 4 different CT scanners. We found that the radiation dose for the same CT technique can vary by as much as a factor of 3 when 1 mm slices are used. CT dose variation was greatest for thin slices, making these observations particularly important for high-resolution CT. Our measurement technique used standard quality assurance equipment available in most radiology departments. Use of these measurements to assess the radiation dose from different CT scanners is an easily performed technique that may allow a decrease in radiation exposure in departments by choosing the most appropriate intra departmental CT scanner for specific indications. PMID- 14561911 TI - Linking information on the World Wide Web. AB - The uniform resource locator (URL) is the key element of the World Wide Web, connecting widely separate sources of information. Logically constructed URLs are important in helping users find desired information. URLs can deceive users. Cyber squatting and other practices that manipulate URLs often lead the user to unwanted information. Critical to future scholarship and education is access to full text articles online. This will require consistent and well-constructed URLs. PMID- 14561912 TI - Post-traumatic pericardial calcification. AB - Trauma is a cause of calcific constrictive pericarditis, but less commonly may cause a localized mass. We report such a case in which the mass impinged on the right heart resulting in severe systemic venous stasis. PMID- 14561913 TI - Rapid and irreversible cystic change of pulmonary tuberculosis in an immunocompetent adult. AB - This case report illustrates rapid and irreversible cystic change associated with disseminated pulmonary tuberculosis in an immunocompetent adult. In this patient, cystic lesions persisted after successful treatment of Mycobacterium tuberculosis and resulted in spontaneous pneumothorax with persistent air leak. PMID- 14561914 TI - Variability of gray and white matter during normal development: a voxel-based MRI analysis. AB - This study was aimed at investigating regional and local variability of brain tissue during normal human brain development. We investigated high-resolution MR imaging data using SPM99 (Wellcome Department, University College London, UK). A pixel-wise variation coefficient of gray and white matter was calculated to visualize local variability. Data from 200 normal children (5-18 years) were analyzed. We found a profound interaction between variability of brain structures and normal development in both gray and white matter. Variability in gray and white matter shows regionally specific, age-related variations, possibly offering a new tool for the assessment of subtle brain abnormalities. Our results emphasize the necessity to take this variability into account when planning pediatric neuroimaging studies. PMID- 14561915 TI - Olfactory neurons in bax knockout mice are protected from bulbectomy-induced apoptosis. AB - Surgical ablation of the olfactory bulb (bulbectomy) triggers a massive wave of apoptosis in mature olfactory sensory neurons within the olfactory epithelium. The aim of the current study was to determine if this process is dependent on expression of the pro-apoptotic protein Bax. Immunohistochemical detection of caspase-3 activation and olfactory epithelial thickness was used to demonstrate and quantify neuronal apoptosis in bax knockout and wild type mice, following bulbectomy. Caspase-3 activation and epithelial thinning were both reduced in the bax knockout mouse compared to the wild type mouse, at least up to 9 days post bulbectomy, indicating that apoptosis was inhibited not just delayed. This study demonstrates that Bax plays a major role in olfactory neuron apoptosis following surgical deafferentation. PMID- 14561916 TI - Reverse sequencing syllables of spoken words activates primary visual cortex. AB - Using fMRI, we investigated the neural correlates for sequencing the individual syllables of spoken words in reverse order. The comparison of this task to a control task requiring subjects to repeat identical syllables given acoustically revealed the activation of the primary visual cortex. Because one syllable is generally expressed by one kana character (Japanese phonogram), most subjects used a strategy in which the kana character string corresponding to the word was imagined visually and then read mentally in reverse order to perform the task effectively. Such strategy was not used during a control condition. These results suggest that the primary visual cortex plays a role in the generation of an imagined string. PMID- 14561918 TI - Is the velocity-curvature relationship disrupted in apraxic patients? AB - Velocity and curvature of human movements are linked by a proportionality relationship (power-law) whose origin has been attributed either to functional properties of cortical areas or to peripheral constraints. 3D movements made by apraxic patients show a time-shift between velocity and curvature which has been considered as a disruption of the power-law, supporting the central hypothesis. We analysed the power-law in 2D drawing-like movements in healthy subjects and apraxic patients (correlation and cross-correlation analyses). The power-law remained preserved in apraxic patients, suggesting that the velocity-curvature relationship is not globally disrupted and thus that the power-law cannot be only attributed to central planning mechanisms in those associative brain areas injured in apraxic patients. PMID- 14561917 TI - Corticospinal excitability changes following prolonged muscle tendon vibration. AB - The present experiment addressed the time course of corticospinal excitability changes following interventional muscle tendon vibration. Using transcranial magnetic stimulation, motor evoked potentials of the flexor carpi radialis and extensor carpi radialis brevis muscle were recorded for a period of 60 min after cessation of vibration (80 Hz, 0.5 mm, 30 min) to the distal wrist flexor tendons. A delayed corticospinal excitability increase in both the vibrated and non-vibrated antagonistic muscle was observed, with lasting levels of facilitation for the latter. No changes were observed following interventional cutaneous vibration. These results underscore a facilitatory influence of prolonged Ia-afferent activation on corticospinal excitability. Findings are discussed in light of recent advances in promoting motor recovery after brain injury by somatosensory stimulation. PMID- 14561919 TI - Betaxolol attenuates retinal ischemia/reperfusion damage in the rat. AB - This study was performed to elucidate the protection afforded by post-treatment with Betoptic (0.25% betaxolol) against neuronal cell damage after ischemia/reperfusion insult in rats. Betaxolol was applied topically after the start of reperfusion and its effect was evaluated by morphometry and choline acetyltransferase immunoreactivity of retinas at 7 days after reperfusion. In non treated eyes, the thickness of the inner plexiform layer decreased markedly after a reperfusion period of both 3 and 7 days. However, when eyes were treated with betaxolol after ischemia/reperfusion injury, both the reduction of the inner plexiform layer thickness and the retinal choline acetyltransferase immunoreactivity were significantly attenuated. These findings suggest that betaxolol is an efficient neuroprotective agent and prevents the retinal cell damage induced by ischemic injury in rats. PMID- 14561920 TI - The pattern and distribution of calcitonin gene-related peptide (CGRP) terminals in the rat dorsal following neonatal peripheral inflammation. AB - Neonatal peripheral inflammation has been shown to alter the neural circuitry of the spinal cord in adult rats. However, the temporal and spatial changes in the distribution of primary afferent terminals immediately following a neonatal inflammatory stimulus remains unclear. In the present study we found that intraplantar injection of complete Freund's adjuvant (CFA) or saline alone on postnatal day 1 (P1) causes CGRP immunoreactivity (CGRP-Ir) to gradually increase from P6 to P15 in laminae I and II, and return to baseline at P22. In laminae III and IV, CGRP-Ir markedly increased beginning at P6, and remained elevated thereafter. CGRP-Ir in lamina V remained unchanged throughout the observation period. These findings show that intraplantar CFA induces CGRP-fiber sprouting in laminae III and IV, but not in laminae I, II or V. We suggest that neonatal inflammation causes changes in the neural circuitry pattern in various regions of the dorsal horn during the critical neonatal development period in rats. PMID- 14561921 TI - Involvement of apolipoprotein E in glioblastoma: immunohistochemistry and clinical outcome. AB - We hypothesised that apolipoprotein E (apoE) influences brain tumours by delivery of lipids to tumour cells and by analogy with other brain insults. APOE gene analysis was performed for 126 glioblastomas, the commonest primary brain tumour. Neither APOE epsilon2 nor epsilon4 alleles were significantly associated with differences in post-operative survival. However, there was apoE immunoreactivity of tumour cells, macrophages in areas of necrosis and astrocytes nearby. The immunohistochemistry findings support the hypothesis that apoE is involved in the delivery of lipids to tumour cells and in the recycling of lipids by macrophages in necrotic areas, raising the possibility that apoE-mediated lipid transport may represent a new therapeutic target in brain tumours. PMID- 14561922 TI - COX-2-deficient mice are less prone to MPTP-neurotoxicity than wild-type mice. AB - The primary lesion in Parkinson's disease is the death of dopaminergic neurons in the substantia nigra. The role of cyclooxygenase (COX)-2 in the etiology of Parkinson's disease was explored using COX-2 gene knockout mice. Mortality after injection of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP, a chemical known to cause parkinsonism in humans) in heterozygous COX-2-deficient mice was lower than that in wild-type mice. The number of tyrosine hydroxylase immunoreactive neurons in the substantia nigra pars compacta of MPTP-treated wild-type mice declined to a greater extent than in heterozygous mice. Inhibition of COX-2 protein expression decreased the lesion caused by MPTP and protected the dopaminergic neurons in substantia nigra pars compacta. This result suggested that inhibition of COX-2 has potential therapeutic implications. PMID- 14561923 TI - Functional expression and properties of a nicotinic alpha9/5-HT3A chimeric receptor. AB - We describe the functional properties of a nicotinic alpha9/serotonin subtype 3A (5HT3A) chimeric receptor expressed in Xenopus laevis oocytes. The chimera preserved ligand-binding properties of alpha9 and channel properties of 5HT3A. Thus, it responded to acetylcholine in a concentration-dependent manner with an EC50 of 70 microM but not to serotonin. It was blocked by methyllycaconitine, strychnine, atropine and nicotine, with the same rank order of potency as alpha9 receptors. The current-voltage relationship of currents through the alpha9/5HT3A chimera was similar to that of the 5HT3A receptors. These results are an evidence of functional coupling between the ligand-binding and the channel domains of the chimeric receptor. PMID- 14561924 TI - Differential effects of 9-cis, 13-cis and all-trans retinoic acids on the neuronal differentiation of human neuroblastoma cells. AB - A human neuroblastoma cell line IMR-32 was used as an in vitro model to examine three naturally occurring retinoic acid (RA) isomers, 9-cis (9c), 13-cis (13c) and all-trans (AT) RA, in mediating growth differentiation and neuronal differentiation. All RA isomers inhibited cellular proliferation, with 13c-RA being most effective. Cyclic AMP-responsive-element-binding-protein (CREB) was activated during RA treatment. AT-RA was a better differentiating agent in inducing the highest expression of the neurotrophic factor receptor TrkA. After prolonged RA treatment, the expression of RA receptors (RARs) was comparable for the three isomers, but retinoid X receptors (RXRs) were differentially regulated. These results imply that distinctive molecular pathways might be involved in the in vitro differentiation of neuroblastoma with different RA isomers. PMID- 14561925 TI - Falcarindiol inhibits nitric oxide-mediated neuronal death in lipopolysaccharide treated organotypic hippocampal cultures. AB - Excessive nitric oxide (NO) release from activated microglia has a predominant role in neuronal death. This study investigated the effect of falcarindiol, which was isolated from Cnidium officinale Makino, on the NO-mediated neuronal death in lipopolysaccharide (LPS)-treated organotypic hippocampal cultures. Falcarindiol dose-dependently reduced inducible NO synthase (iNOS)-mediated NO production without cytotoxic effects on LPS-activated BV-2 and microglia. Predictably, falcarindiol inhibited neuronal death by reducing NO production in the LPS treated organotypic hippocampal cultures. N-monomethyl-L-arginine (NMMA), an iNOS inhibitor, also inhibited neuronal death at 500 microM. In contrast, massive neuronal death was induced by excessive NO production in the LPS-treated alone cultures. These results suggest that excessive NO production plays an important role in the neurotoxic effect, and falcarindiol is a potential inhibitor in NO mediated neuronal death. PMID- 14561926 TI - Inhibitory effect of the DA uptake blocker GBR 12909 on sodium channels of hippocampal neurons. AB - The effect of the selective dopamine uptake inhibitor GBR 12909 on TTX-sensitive sodium channels of cultured hippocampal neurons was investigated using whole cell patch-clamp technique. GBR 12909 dose-dependently inhibited sodium currents evoked by trains of depolarizing pulses with an IC50 of 6.3 microM. A weaker inhibition (IC50 = 17-35 microM) could be observed when currents were evoked by either single pulse depolarization or from hyperpolarized holding membrane potential. These data indicate that the extent of inhibition caused by GBR 12909 depends on the physiological activity pattern of neurons. Our results suggest that caution is needed for the interpretation of data when GBR 12909 is used for the inhibition of dopamine uptake at concentrations above the submicromolar range. PMID- 14561927 TI - Adenoviral vector gene delivery via the round window membrane in guinea pigs. AB - We have found that damage from a local anesthetic solution containing phenol permitted beta-galactosidase (beta-gal) gene delivery to the guinea pig inner ear via the round window membrane (RWM). RWM damage was evident as degeneration of the outer epithelium. After adenovirus lacZ vector was applied to the damaged RWM, immunohistochemistry showed strong beta-gal expression in the RWM, mesothelial cells, organ of Corti, spiral limbus, spiral ligament and spiral ganglion. In the vestibular labyrinth, expression was seen in the sensory and supporting cells, transitional cells, and the dark-cell area. Thus, adenovirus can transfect a variety of inner ear cells in the guinea pig through a damaged RWM. PMID- 14561928 TI - A non-invasive method for observing hippocampal function. AB - A non-invasive method for observing the functioning of the hippocampus could be invaluable in understanding the role of hippocampal abnormalities in many brain disorders. Transverse patterning, a hippocampal-dependent memory task, was used in an attempt to study the functioning hippocampus. Subjects performed transverse patterning while whole-head MEG data were collected. The MEG data were analyzed using a spatial-temporal multiple-dipole approach. Controls showed right hippocampal activation. Patients with unilateral hippocampal damage showed activation in undamaged hippocampus. MEG during transverse patterning performance is a promising, non-invasive tool for assessing hippocampal function. PMID- 14561929 TI - Functional assessment of Broca's area using near infrared spectroscopy in humans. AB - We used near-infrared spectroscopy (NIRS) to compare functional hemoglobin concentration changes (delta[oxy-Hb] and delta[deoxy-Hb]) over human language and motor cortices. Eight subjects performed finger opposition, tongue movement, and covert visual object naming in an interleaved block paradigm design. NIRS revealed paradigm specific patterns of delta[oxy-Hb] and delta[deoxy-Hb] providing cortical localization of each function. During each task, significant response overlap was observed when comparing the [oxy-Hb] signals, whereas delta[deoxy-Hb] seemed more localized. Furthermore, by applying magnitude and time to significance measures to the delta[deoxy-Hb] response profile, Broca's area was easily distinguished from neighboring tongue (and hand) motor representation. Delta[oxy-Hb] did not provide this level of specificity. These findings suggest delta[deoxy-Hb] as the preferential NIRS parameter to map language cortices. PMID- 14561930 TI - Cells over-expressing EAAT2 protect motoneurons from excitotoxic death in vitro. AB - Amyotrophic lateral sclerosis is an incurable disease in which cerebral and spinal motoneurons degenerate, causing paralysis and death within 2-5 years. One of the pathogenic factors of motoneuron death is a chronic excess of glutamate, which exceeds its removal by astrocytes, i.e. excitotoxicity. Extra glutamate uptake in the spinal cord may slow down or prevent motoneuron death. We have engineered cells over-expressing the main glutamate transporter and tested their potential to rescue motoneurons exposed to high levels of glutamate in vitro. The engineered cells protected motoneurons in a motoneuron-astrocyte co-culture at glutamate concentrations when astrocytes were no longer capable of removing glutamate. This suggests that engineered cells, introduced into the spinal column, can help remove glutamate, thereby preventing motoneuron death. PMID- 14561931 TI - Mechanisms of the anti-ischemic effect of vagus nerve stimulation in the gerbil hippocampus. AB - The neuroprotective mechanisms of cervical vagus nerve stimulation (VNS) in transient ischemia were investigated. Left VNS (0.4 mA, 40 Hz) was performed during 5 min ischemia in gerbils. About 50% of the hippocampal neurons were rescued from ischemic insult by VNS, and this effect was prevented by transection of the vagus nerve centrally to the site of cervical stimulation. VNS significantly attenuated both ischemia-induced glutamate release and transient increase of hippocampal blood flow during reperfusion. Hyperemia as well as excessive glutamate release after ischemia is regarded as an important factor in ischemic brain damage as it leads to generate considerable reactive oxygen species. Thus, VNS might protect neurons from ischemia-induced glutamate excitotoxicity and reperfusion injury via the afferent path-way of the vagus. PMID- 14561932 TI - Genetic influences on frontal brain function: WCST performance in twins. AB - The Wisconsin Card Sorting Test (WCST) is one of the most widely used assessments of executive functioning related to prefrontal cortex. However, little is known about genetic and environmental determinants of individual differences in WCST performance. This study assessed heritability of standard WCST scores in a sample of 168 young female twins including 58 monozygotic and 25 dizygotic pairs. Several WSCT indices, including the number and percentage of errors, the number of perseverative responses, and the number and percent of perseverative errors, showed significant heritability ranging from 37 to 46%. The results suggest that selected aspects of frontal executive functioning measured by the WCST are moderately influenced by genetic factors. PMID- 14561933 TI - Impaired spatial reference memory in aromatase-deficient (ArKO) mice. AB - Previous studies indicate an important role for estrogen in memory and learning. Aromatase-knockout (ArKO) mice are unable to produce estrogen because they lack a functional Cyp 19 gene that encodes for aromatase, the enzyme that converts testosterone into estrogen. Using a Y-maze test for short-term spatial reference memory, we found that both male and female ArKO mice performed significantly worse than wildtype controls. Gonadectomy reduced Y-maze responses in male and female wildtype controls, but had no effect in ArKO mice. After gonadectomy, there was no significant difference between wildtype and ArKO mice. For the first time using ArKO mice, our findings confirm the importance of estrogen in memory in both males and females. PMID- 14561935 TI - Memory formation requires p38MAPK activity in the rat hippocampus. AB - Using the specific inhibitor of p38MAPK, SB203580, we show a direct involvement of this protein kinase in short- and long-term memory. When given into the CA1 region of the rat dorsal hippocampus immediately, but not 30 or 120 min after training in a one-trial inhibitory avoidance task, SB203580 blocked short- and long-term memory formation. The SB203580 inactive analog, SB202474, had no effect whatsoever. Learning of the avoidance task was accompanied by an immediate and transient increase in hippocampal p38MAPK phosphorylation. No change in p38MAPK phosphorylation was detected in control animals that only received the electric foot-shock associated with the learning paradigm. Therefore, formation of short and long-term memory for inhibitory avoidance requires p38MAPK activation in the rat hippocampus. PMID- 14561934 TI - Neural correlates of working memory in pure and polyvalent ecstasy (MDMA) users. AB - Poor cognitive performance in ecstasy (3,4-methylenedioxymethamphetamine; MDMA) users has been related to the well-recognized neurotoxic effects of the drug upon central serotonergic and possibly also dopaminergic systems. However, concomitant use of other drugs has been a critical confound in most investigations. In this study we used an n-back task and fMRI to investigate working memory performance and related cerebral activation in eight, currently abstinent pure MDMA users and two matched groups of polyvalent MDMA users and non-users. Pure MDMA users presented lower activations than controls and/or polyvalent users, most notably in inferior temporal regions, the angular gyrus and the striate cortex, whereas polyvalent users did not differ from controls. Our results suggest that altered brain activation patterns during cognitive processing in ecstasy users may be mainly associated with prior MDMA use. Concomitant use of other drugs may modify this effect. PMID- 14561936 TI - Low cortisol levels and the balance between punishment sensitivity and reward dependency. AB - The neuro-endocrinological basis of psychopathy, a disorder characterized by lack of fear, is relatively unknown. However, low levels of cortisol may result in fearlessness and have been observed in individuals with psychopathic tendencies. Low fear models state that psychopaths are not motivated to avoid punishment, especially when reward is pending. In agreement, disadvantageous decision making on the IOWA gambling task in psychopaths and psychopathic analogous, indicates low punishment sensitivity and high reward dependency. Here, it was investigated whether low basal cortisol levels predicted the balance between punishment sensitivity and reward dependency in the same manner. As hypothesized, the most disadvantageous pattern of decision making was found in the subjects with the lowest cortisol levels. These findings suggest that low levels of cortisol may set the balance between the sensitivity for punishment and reward dependency towards a pre-disposition for psychopathy. PMID- 14561937 TI - Neuromedin U facilitates oxytocin release from the pituitary via beta adrenoceptors. AB - Neuromedin U activates noradrenergic neurones in the medulla oblongata and oxytocin neurones in the hypothalamus. Here we examined roles of noradrenergic transmission in oxytocin release from the pituitary after intracerebroventricular administration of neuromedin U in rats. Neuromedin U administration facilitated noradrenaline release in the supraoptic nucleus. Administration of a beta1 adrenoceptor antagonist, metoprolol, or a beta2 antagonist, ICI 118551 but not an alpha1 antagonist, benoxathian, reduced increases in plasma oxytocin concentrations observed after administration of neuromedin U, but plasma ACTH concentrations were not significantly changed. All theses data suggest that neuromedin U stimulates oxytocin release from the pituitary, at least in part, via activation of beta adrenoceptors. PMID- 14561939 TI - The articulatory loop: study of the subcortical connectivity by electrostimulation. AB - Although a cortical network involving Broca's area and the supramarginal gyrus (SMG) was widely studied using neurofunctional imaging, the functional connectivity underlying this so-called articulatory loop remains poorly documented. We describe a patient operated on for a glioma invading the left parietal operculum, using intraoperative electrical functional mapping under local anesthesia. Following the identification of cortical language sites within Broca's area and SMG, the subcortical pathways connecting these regions were detected and preserved during the resection. Postoperatively, the patient presented a slight dysarthria, then recovered. This is the first report of direct tracking of the subcortical connectivity underlying the fronto-parietal articulatory loop, allowing to better understand the pathophysiology of this network and the consequences of its damage. PMID- 14561938 TI - Excitatory effects of 5-hydroxytryptamine on the medial vestibular nuclear neuron via the 5-HT2 receptor. AB - This study was designed to investigate the mechanisms of action of the 5-HT2 receptor on the spontaneous electrical activity and potassium currents of the rat medial vestibular nuclear neurons using whole-cell patch clamp recordings. The spike width of spontaneous action potential was not affected by 5-alpha methylhydroxytryptamine. The spike frequency and resting membrane potential was increased by 5-alpha-methylhydroxytryptamine. The amplitude of afterhyperpolarization was decreased by 5-alpha-methylhydroxy-tryptamine. The peak current of the potassium currents of the neuron treated with 5-alpha methylhydroxytryptamine was decreased. After blockade of calcium-dependent potassium currents, 5-alpha-methylhydroxytryptamine did not inhibit potassium currents. These results suggest 5-alpha-methylhydroxytryptamine increases spontaneous firing of the medial vestibular nuclear neurons by inhibiting calcium dependent potassium currents. PMID- 14561940 TI - Endothelin, hypercholesterolemia and atherosclerosis. PMID- 14561941 TI - Endothelins and coronary vascular biology. PMID- 14561942 TI - Endothelin in neurohormonal activation in heart failure. PMID- 14561943 TI - Reactive hyperemia following coronary balloon angioplasty, but not dipyridamole induced hyperemia, predicts resolution of exercise-induced ST-segment depression. AB - OBJECTIVES: To characterize delayed restoration of coronary blood flow following successful percutaneous transluminal coronary angioplasty (PTCA). BACKGROUND: Delayed restoration of coronary blood flow following successful PTCA is common and likely the result of multiple factors. Temporary myocardial ischemia and dipyridamole administration both result in increased coronary blood flow, but by different mechanisms. The relationship between these phenomena and exercise induced ST-segment depression after PTCA was investigated to determine if any correlation existed. METHODS: Forty consecutive patients with single-vessel coronary artery disease underwent treadmill exercise testing before and after PTCA. The percentage change in coronary blood flow before and after 90 s balloon inflation was assessed. After a new steady state had been reached, dipyridamole was infused and changes in coronary blood flow were again determined. The relationship between changes in coronary blood flow and the presence of ST segment depression during exercise testing after PTCA was determined. RESULTS: Peak coronary blood flow induced by reactive hyperemia was significantly greater than that in the steady state after balloon inflation (48.5+/-38.8 compared with 15.1+/-13.2 ml/min, P<0.0001). Dipyridamole administration also resulted in significant increases in coronary blood flow (15.1+/-13.2 ml/min compared with 31.0+/-24.9 ml/min, P<0.0001). ST-segment depression after PTCA was significantly less than before (0.10+/-0.07 mV compared with 0.19+/-0.08 mV, P<0.001). Further, reactive hyperemia, but not dipyridamole-induced hyperemia, correlated with attenuation of exercise-induced ST-segment depression after PTCA (r=0.62, P<0.0001). CONCLUSIONS: Reactive hyperemia following temporary coronary occlusion recreates local conditions associated with delayed resolution of myocardial ischemia following successful PTCA. Further, this phenomenon appears to be distinct from changes in coronary blood flow induced by dipyridamole. PMID- 14561944 TI - Improvement and factors associated with improvement in quality of life during 10 years after coronary artery bypass grafting. AB - AIM: To describe (1) the improvement in various aspects of quality of life (QoL) and (2) predictors of improvement, during 10 years after coronary artery bypass grafting (CABG). PATIENTS AND METHODS: All patients who underwent CABG in western Sweden between June 1988 and June 1991 without simultaneous valve surgery and with no previous CABG were approached with an inquiry prior to and 5 and 10 years after the operation. QoL was measured with three different instruments: (1) Nottingham health profile (NHP), (2) psychological general well-being index (PGWBI) and (3) physical activity score (PAS). RESULTS: There was a significant improvement in QoL with all three instruments from before to 10 years after the operation. The mean improvements +/-SD were for NHP, - 4.2+/-17.0 (P<0.0001), for PGWBI, +9.7+/-17.6 (P<0.0001) and for PAS, -0.96+/-1.23 (P<0.0001). However, there was also a deterioration with all three instruments between 5 and 10 years after surgery. The mean deteriorations +/-SD were for NHP, +4.4+/-12.8 (P<0.0001), for PGWBI, -4.6+/-14.8 (P<0.0001) and for PAS, +0.44+/-0.94 (P<0.0001). Independent predictors for an improvement in QoL with at least one of the instruments were low preoperative QoL, a younger age, being a man, high functional class (New York Heart Association), no hypertension, proximal left anterior descending coronary artery stenosis, short extracorporeal circulation time, use of internal mammary artery and a short postoperative time in the intensive care unit. CONCLUSION: There is a higher estimated QoL 10 years after CABG than before, despite the fact that the patients are 10 years older. However, there is also a deterioration in QoL between 5 and 10 years after surgery. Predictors of improvement during the 10 years included age, sex, previous history, localization of stenosis, type of graft and preoperative and postoperative factors. PMID- 14561945 TI - Psychotropic drug-induced weight gain alleviated with orlistat: a case series. AB - Weight gain is a common side effect associated with antidepressant, anxiolytic, and antipsychotic drug use. Obesity is a risk factor for several other disorders, including hypertension, diabetes, and coronary artery disease. To date, there have been few safe, well-tolerated, and effective pharmacological agents available to alleviate weight gain in general, and virtually no studies specific to psychiatric drug-induced weight gain. This case series looks at the use of orlistat, a reversible inhibitor of lipases approved by the US Food and Drug Administration for obesity management, naturalistically in 13 patients with weight gain secondary to psychotropic drug use. The results showed that orlistat, administered in 3 daily doses with meals, was safe, well-tolerated, and effective, resulting in an average weight loss of 35% during an acute treatment period of about 3 months. PMID- 14561946 TI - Pharmacokinetics and drug interactions of the sedative hypnotics. AB - Sleep complaints are common among patients and may be associated with mild to severe clinical symptoms. The use of hypnotic agents to improve the onset and quality of sleep has a history dating back centuries. Currently, several benzodiazepines and nonbenzodiazepines are widely used as sedative hypnotic drugs. The pharmacokinetic and pharmacodynamic profiles of these agents vary substantially, allowing them to be chosen for different clinical situations and tailored to specific sleep complaints. The short-acting benzodiazepines and benzodiazepine agonists have become the most widely prescribed sedative hypnotics because of their short to ultrashort half-life, and low tendency to impair cognitive and psychomotor performance on the day following nighttime use. In the current review, we summarize the pharmacokinetic and drug interaction profiles of the Food and Drug Administration-approved benzodiazepine sedative hypnotics: estazolam, flurazepam, quazepam, temazepam, and triazolam. In addition, alprazolam is included because of its popularity for this use, and 2 benzodiazepine receptor agonists, zolpidem and zaleplon, are reviewed. The presented data will provide clinicians with insight into the pharmacokinetic profiles of several popular sedative hypnotic agents, and provide guidelines to help them recognize or avoid drug interactions. PMID- 14561947 TI - Evidence for the treatment of borderline personality disorder. AB - The treatment of patients with borderline personality disorder (BPD) is notoriously difficult. Psychotherapeutic and pharmacological strategies have been investigated, and a few have shown promise. Dialectical behavior therapy (DBT) may be helpful in decreasing suicidal behavior and improving symptomatology, although the data are not strong, and the actual "type" of DBT employed may influence the outcomes. Although there are a variety of studies investigating a number of different medications for the treatment of BPD, there are few randomized, double-blind, placebo-controlled trials. Furthermore, the evidence base is limited by small sample size and variability in inclusion criteria and outcome measures among the different studies. Further study is needed with larger, randomized, placebo-controlled trials to clearly demonstrate benefit of any pharmacotherapy. To date, there is suggestion from some studies that neuroleptics, carbamazepine, monoamine oxidase inhibitors, and selective serotonin reuptake inhibitors may be effective, and there is evidence that benzodiazepines are detrimental. The role of electroconvulsive therapy remains unclear. PMID- 14561948 TI - New approaches to modeling bipolar disorder. AB - Modeling in animals is an invaluable tool in exploring the pathophysiology of human diseases and developing better therapies. Models can be generated using a variety of pharmacological, behavioral, and genetic approaches, but they all require extensive subsequent validation. Ideally, validation should be based on the following 3 axes: face validity (commonalties between the behavioral features of the model and of the human disorder being modeled), predictive validity (the specificity and degree to which drugs that are effective in humans have a corresponding effect in the model), and construct validity (a possible common mechanistic theory that can explain both the model and the human disorder being modeled). Most existing models for psychiatric disorders were developed from a face validity starting point, wherein a researcher noticed the appearance of a rodent behavior that was similar to a human pathological behavior and subsequently undertook investigations of predictive and construct validity. Some representative models developed in this manner include the hyperactivity (spontaneous or pharmacologically induced), sensitization, and sleep-deprivation models. In this review, we critically appraise the existing animal models for bipolar disorder, emphasizing their strengths and limitations. Furthermore, we discuss the technological advances that have led to an increased awareness of the roles of signal transduction pathways and neurotrophic cascades in the pathophysiology and treatment of bipolar disorder. New construct validity-driven models focusing on signaling pathways include models based on perturbations of G proteins, phosphoinositide signaling, and mitogen-activated protein (MAP) kinase cascades. These new models hold much promise in delineating the underlying pathophysiology of bipolar disorder and for the development of novel, improved therapeutics. Psychopharmacology Bulletin. PMID- 14561949 TI - Does isotretinoin cause depression and suicide? AB - Isotretinoin is a retinoid that is approved for the treatment of cystic acne. There has been a growing interest in the possible relationship between isotretinoin use and increased risk of depression and suicide. The issue, however, remains controversial. This review examines the available evidence on this association, including published case reports, studies of challenge rechallenge, reports to government agencies of isotretinoin-related adverse events involving suicide and depression, and possible biological mechanisms of action. Given the evidence to date, clinicians are advised to counsel their patients about the risk of depression and suicide as possible side effects of isotretinoin use, and to administer self-report questionnaires during treatment to screen for the development of depression. Further epidemiological and neurobiological studies are needed to assess the possible relationship between isotretinoin administration and depression and suicide. PMID- 14561950 TI - An intent-to-treat method for enhancing analysis of clinical trials with rescue medication: a mixed-model approach. AB - An intent-to-treat (IT) analysis includes all observed data, even those obtained after the primary treatment assignment is changed or rescue treatment is substituted. The so-called "pragmatic" approach accounts for this type of treatment change or rescue treatment. The advantage of the pragmatic approach to IT analysis is that the timing and duration of the rescue treatment can be included in the analysis with mixed-effect modeling, which estimates the difference between the randomized treatment arms while adjusting for the use (timing and duration) of the subsequent treatment. We illustrate the inclusion of information on rescue treatment timing and duration in the analysis of a two-arm randomized clinical trial comparing antidepressant medications in an elderly depressed population. In this study, the randomized treatments are found to be similar. This enhanced analysis, where information about rescue treatment is implicitly added in the model, corroborates our previous two-piece spline analysis, where information about rescue treatment was not included. In addition, a significant drug by rescue interaction indicates a differential effect of the timing of rescue treatment between the two treatments. This analytic approach illustrates the importance of collecting and incorporating data after rescue, including the timing of rescue. PMID- 14561952 TI - Overview of the safety of citalopram. AB - Citalopram is a highly selective serotonin reuptake inhibitor (SSRI) that has been prescribed to >30 million patients in >70 countries. The purpose of this focused overview is to summarize the data from well-controlled clinical trials and published literature relative to the safety of citalopram in patients with depression and depressive symptoms. This overview is based mainly on 3 sources: (1) data from clinical trials sponsored by Forest Laboratories, (2) published clinical studies, and (3) case reports. Both pharmacokinetic and pharmacodynamic interactions were scrutinized, as were data on special populations and safety concerns. The available data suggest that citalopram 20-60 mg once daily is safe for patients with depression. Few drugs appear to interact with citalopram in a clinically meaningful way. Well-designed short- and long-term trials demonstrate an overall safety/side effect profile consistent with other SSRIs. The more frequent adverse events (nausea, somnolence, dry mouth, increased sweating) are mainly transient, mostly mild to moderate in severity, and observed consistently across studies at rates similar to other SSRIs. Analysis of laboratory values, ECG, and vital signs revealed no unusual findings. Only a small, clinically unimportant reduction in heart rate was observed, similar to that seen with other SSRIs. Citalopram treatment did not increase risk of suicide, overdose, seizure, or arrhythmia. Thus, the pharmacodynamic, pharmacokinetic, and safety profiles of citalopram demonstrate that it is safe for use in adults with depression and depressive symptoms, including the elderly and patients with mild to moderate renal and hepatic disease. PMID- 14561951 TI - Smoking abstinence impairs time estimation accuracy in cigarette smokers. AB - Time estimation is used as an index of attention processes and may be sensitive to self-administration and withdrawal of psychoactive drugs such as nicotine, the primary addictive ingredient in tobacco. To test this hypothesis, 22 nonsmokers (12 male, 10 female) and 20 daily cigarette smokers (12 male, 8 female) were asked to estimate the duration of a 45-second period of time in a laboratory setting. Smokers participated in two sessions: once after smoking ad-lib and once after objectively confirmed 24-hour smoking abstinence. In smokers, time estimation accuracy was impaired after smoking abstinence compared to accuracy after ad-lib smoking (P<.01). Relative to nonsmokers, smokers' time estimation accuracy differed significantly after the abstinence period only (P=.05). Smokers reported feeling more stressed and unable to focus after abstinence relative to nonsmokers (P<.05). No gender differences were observed on any outcome measures. These results suggest that, in a controlled laboratory setting, smoking abstinence has a negative impact on time perception, which may contribute to the performance decrements and discomfort that smokers report during an attempt to quit. PMID- 14561953 TI - Preventing skin cancer: findings of the Task Force on Community Preventive Services On reducing Exposure to Ultraviolet Light. AB - Rates of skin cancer, the most common cancer in the United States, are increasing. The most preventable risk factor for skin cancer is unprotected ultraviolet (UV) exposure. Seeking to identify effective approaches to reducing the incidence of skin cancer by improving individual and community efforts to reduce unprotected UV exposure, the Task Force on Community Preventive Services conducted systematic reviews of community interventions to reduce exposure to ultraviolet light and increase protective behaviors. The Task Force found sufficient evidence to recommend two interventions that are based on improvements in sun protective or "covering-up" behavior (wearing protective clothing including long-sleeved clothing or hats): educational and policy approaches in two settings--primary schools and recreational or tourism sites. They found insufficient evidence to determine the effectiveness of a range of other population-based interventions and recommended additional research in these areas: educational and policy approaches in child care centers, secondary schools and colleges, recreational or tourism sites for children, and workplaces; interventions conducted in health-care settings and targeted to both providers and children's parents or caregivers; media campaigns alone; and community wide multicomponent interventions. This report also presents additional information regarding the recommended community interventions, briefly describes how the reviews were conducted, provides resources for further information, and provides information that can help in applying the interventions locally. The U.S. Preventive Services Task Force conducted a systematic review of counseling by primary care clinicians to prevent skin cancer (CDC. Counseling to prevent skin cancer: recommendation and rationale of the U.S. Preventive Services Task Force. MMWR 2003;52[No. RR-15]:13-17), which is also included in this issue, the first jointly released findings from the Task Force on Community Preventive Services and the U.S. Preventive Services Task Force. PMID- 14561954 TI - Counseling to prevent skin cancer: recommendations and rationale of the U.S. Preventive Services Task Force. AB - This statement summarizes the current U.S. Preventive Services Task Force (USPSTF) recommendation on counseling to prevent skin cancer and the supporting scientific evidence, and updates the 1996 recommendation contained in the Guide to Clinical Preventive Services, Second Edition (U.S. Preventive Services Task Force. Screening for skin cancer. In: Guide to clinical preventive services, 2nd ed. Washington DC: US Department of Health and Human Services, Office of Disease Prevention and Health Promotion, 1996:141-52). The USPSTF finds insufficient evidence to recommend for or against routine counseling by primary care clinicians to prevent skin cancer. Although counseling parents may increase children's use of sunscreen, the USPSTF found little evidence to determine the effects of counseling on the sun protection behaviors of adults. These behaviors include wearing protective clothing, reducing excessive sun exposure, avoiding sun lamps and tanning beds, or practicing skin self-examination. The USPSTF, an independent panel of private sector experts in primary care and prevention, systematically reviews the evidence of effectiveness of a wide range of clinical preventive services, including screening tests, counseling, and chemoprevention. Members of the USPSTF represent the fields of family medicine, gerontology, obstetrics/gynecology, pediatrics, nursing, and prevention research. PMID- 14561955 TI - Infants tested for hearing loss--United States, 1999-2001. AB - Hearing loss (HL) occurs in one to three of 1,000 live births annually and, when left undetected, can result in developmental delays. To promote communication from birth, Early Hearing Detection and Intervention (EHDI) programs support early identification of infants with HL. Without EHDI programs, the average age of identification for HL is age 1.5-3.0 years, which is past the start of the critical period for optimal language acquisition. In 2001, a total of 48 states/areas with EHDI tracking and surveillance systems (30 funded by CDC) reported the percentage of newborns screened for HL. This report summarizes the results of an analysis of surveillance data for 1999-2001, which indicate that more infants were screened for HL, received diagnostic audiologic evaluations, and were enrolled in early intervention services in 2001 than in 1999 and 2000. Continued development of EHDI surveillance systems should assist states/areas in providing needed services to children with HL. PMID- 14561956 TI - Prevalence of IgG antibody to SARS-associated coronavirus in animal traders- Guangdong Province, China, 2003. AB - Severe acute respiratory syndrome (SARS) was identified in 2003 as an infectious disease caused by the SARS-associated coronavirus (SARS-CoV), a member of the coronavirus family not observed previously in humans. Because its sequence data differ from that of known human coronaviruses, SARS-CoV is suspected to have crossed the species barrier between an animal host and humans. The SARS outbreak began in China's Guangdong Province, where approximately 1,500 probable cases were identified during November 2002-June 2003. Detection of SARS-like coronavirus has been reported previously in masked palm civets (sometimes called civet cats) and a raccoon dog for sale in a live animal market in Shenzhen municipality. This report summarizes results of an investigation conducted by public health authorities in Guangdong Province, which compared the seroprevalence of SARS-CoV IgG antibody in animal traders (i.e., workers in live animal markets) with that of persons in control groups. The results indicated that 13% of the animal traders, none of whom had SARS diagnosed, had IgG antibody to SARS-CoV, compared with 1%-3% of persons in three control groups. Although the results provide indirect support for the hypothesis of an animal origin for SARS, they also underscore the need for detailed patient histories and more focused animal studies to confirm an animal origin for SARS. PMID- 14561957 TI - Public health and aging: influenza vaccination coverage among adults aged > or =50 years and pneumococcal vaccination coverage among adults aged > or =65 years- United States, 2002. AB - Vaccination of persons at risk for complications from influenza and pneumococcal disease is a key public health strategy in preventing morbidity and mortality in the United States. During the 1990-1999 influenza seasons, approximately 36,000 deaths were attributed annually to influenza infection, with approximately 90% of deaths occurring among adults aged > or =65 years. In 1998, an estimated 3,400 adults aged > or =65 years died as a result of invasive pneumococcal disease. One of the national health objectives for 2010 is to achieve 90% coverage of noninstitutionalized adults aged > or =65 years for both influenza and pneumococcal vaccinations (objective no. 14.29). In 2000, the Advisory Committee on Immunization Practices (ACIP) broadened the universal recommendations for influenza vaccination to include adults aged 50-64 years in addition to adults aged > or =65 years. To assess progress toward achieving the 2010 national health objective and implementing the ACIP recommendations, CDC analyzed data from the 2002 Behavioral Risk Factor Surveillance System (BRFSS). This report summarizes the results of that analysis, which indicate that influenza and pneumococcal vaccination levels among adults aged > or =65 years and influenza vaccination levels among adults aged 50-64 years varied widely among states/areas and racial/ethnic populations. Innovative approaches are needed to increase vaccination coverage, particularly among certain populations. PMID- 14561958 TI - Methicillin-resistant Staphylococcus aureus infections in correctional facilities --Georgia, California, and Texas, 2001-2003. AB - Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) are common in hospitals and nursing homes. Because MRSA is resistant to all commonly prescribed beta-lactam antibiotics (e.g., penicillins and cephalosporins), these infections require treatment with alternative antimicrobial drugs. In addition, because antimicrobial drugs usually must be selected before identifying MRSA as the cause of infection, treatment presents a challenge for clinicians. MRSA has emerged recently as a more frequent cause of skin and soft tissue infections in the community, particularly in correctional facilities such as prisons, jails, and detention centers. This report summarizes recent investigations of MRSA transmission among inmates of correctional facilities in Georgia, California, and Texas. Inadequate personal hygiene, barriers to medical care, and other factors contributed to transmission. Information from these investigations has been used in the development of recently released Federal Bureau of Prisons guidance for control of MRSA, which recommends improvements in inmate hygiene, infection control, and targeted antimicrobial treatment. PMID- 14561959 TI - West Nile virus activity--United States, October 9-15, 2003. AB - This report summarizes West Nile virus (WNV) surveillance data reported to CDC through ArboNET as of 3 a.m., Mountain Daylight Time, October 15, 2003. PMID- 14561961 TI - Endoscopic-assisted repair of subcondylar fractures. AB - OBJECTIVE: To evaluate outcomes of a series of mandibular subcondylar fractures repaired with endoscopic reduction and fixation. Study design Six consecutive subcondylar fractures were treated endoscopically. Intermaxillary fixation was used intraoperatively to aid in fracture reduction. A modified Risdon incision was used to gain access to the lateral ramus, and a modified retractor and endoscope were used for retraction and visualization. Fracture fixation was achieved with a 2-mm titanium plate and screws. Patients were evaluated clinically and radiographically for 6 months and functional, radiographic, and esthetic parameters were assessed at each time period (1, 2, 4, 12, and 24 weeks). RESULTS: All patients demonstrated a stable occlusion in the postoperative period and anatomic alignment of the condyle radiographically. By 1 month, maximum interincisal opening was 42.2 +/- 5.7 mm. There was no joint noise or temporomandibular joint (TMJ) pain postoperatively. Radiographs at each follow up visit indicated the ramus height was maintained in most cases. There was minimal transient facial nerve paresis following surgery. Scar perception was considered acceptable by all patients. Operative times were acceptable as well. CONCLUSION: Endoscopic-assisted repair of subcondylar fractures is an additional tool for management of subcondylar fractures, however there is a steep learning curve based on this study. The technique allows good visualization of the fracture site for reduction through an incision with an acceptable cosmetic result. PMID- 14561962 TI - Complications in bilateral mandibular distraction osteogenesis using internal devices. AB - OBJECTIVE: We sought to evaluate the possibility of distraction osteogenesis as an alternative to conventional bilateral sagittal split osteotomy. Complications (intraoperative, intradistraction, and postdistraction) were evaluated retrospectively. STUDY DESIGN: Seventy consecutive patients (40 males and 30 females, 11.2-37.3 years old; mean, 14.2 years) underwent distraction osteogenesis to lengthen the mandible. The surgical procedure was carried out with the patient under general anesthesia. After the osteotomy was performed, 2 intraoral monodirectional distraction devices were placed on the mandibular cortex in the third molar region. The rate of distraction was 1 mm/day. The different complications encountered during all phases of the distraction procedure were recorded. RESULTS: A total of 28 complications (40%) were recorded. In 10 patients (14.3%), the complications were technique- or device related, or both, and occurred early in the learning period. Five patients (7.1%) had infection occur, and 3 patients (4.3%) had prolonged sensory loss in the distribution of the alveolar nerve. Severe complications occurred in 6 patients (8.6%). Rehospitalization was necessary in 5 patients (7.1%), 4 of whom (5.7% of the series) required further surgery under general anesthesia. CONCLUSION: Distraction osteogenesis can be considered a safe and predictable procedure for lengthening the mandible, with a low incidence of major complications. The infection rate and the incidence of damage to the inferior alveolar nerve (2.1%) are low. Compliance of both patients and parents during the whole treatment period is of the utmost importance. PMID- 14561963 TI - Temporomandibular joint involvement in malignant external otitis. AB - OBJECTIVE: The purpose of this study was to present 6 patients with malignant external otitis (MEO) that resulted in temporomandibular joint (TMJ) involvement and to discuss the incidence, clinical presentation, and treatment modalities. STUDY DESIGN: All patients diagnosed with MEO between 1994 and 2002 were reviewed for cases in which the TMJ was invaded by the infectious process. Only patients in whom TMJ involvement was documented radiographically and in whom the clinical course was well documented were included in this study. RESULTS: MEO was diagnosed in 42 patients over an 8-year period; TMJ involvement was recorded in 6 patients (14%). The medical history revealed controlled type 2 diabetes mellitus in 4 of the 6 patients. All patients reported early ear symptoms, mainly otalgia and otorrhea. Local signs included an ear canal filled with granulation material, edematous overlying skin, and sensitivity to palpation. Cultures taken from the external ear were positive for either Pseudomonas aeruginosa, Staphylococcus epidermidis, Aspergillus, or Proteus mirabilis. TMJ symptoms developed between 1 and 5 months after admission and included painful periauricular swelling and trismus. In 3 patients, healing was uneventful; 3 also died of the disease. CONCLUSIONS: TMJ involvement in MEO is associated with a resistant disease process, often with several recurrences. Prolonged administration of antibiotics is the treatment of choice. Surgical debridement of the TMJ is necessary for the positive identification of the pathogenic organism, in cases of abscess formation, or when osteomyelitic bone destruction of the condyle and glenoid fossa develop. PMID- 14561964 TI - Intraoral melanoma: long-term follow-up and implication for dental clinicians. A case report and literature review. AB - Primary intraoral melanoma is a rare neoplasm with a poor prognosis, accounting for 1% to 8% of all melanoma in Europe and the United States. The incidence (12%) and 5-year survival rate (17.4%) are higher in Japan. We report a case of oral lentiginous melanoma in a Japanese-American man who survived disease-free for more than 5 years after surgery, radiation therapy, and chemotherapy but developed chronic mucositis of the palate under the denture in the primary radiated field. This lesion responded to antifungal therapy. Subsequent multiple biopsies ruled out the recurrence of melanoma but demonstrated prolonged melanocytic hyperplasia and focal epithelial atypia. We reviewed clinical differences in oral melanoma reported in the United States and Japanese literature, and describe the wide variety of oral clinical features of postoperative radiation and chemotherapy, as well as the oral tissue changes caused by denture-induced mucositis and candidiasis in such patients. Dental clinicians should conduct a thorough head, neck, and oral follow-up with increased vigilance in patients with a history of prior cancer. PMID- 14561965 TI - Serum electrolytes in bulimic patients with parotid swellings. AB - OBJECTIVE: The purpose of this study was to determine whether electrolyte levels can be used to diagnose bulimia nervosa (BN). STUDY DESIGN: Four patients with bilateral parotid swellings and BN or suspected BN were examined. Their serum electrolyte levels were retrospectively reviewed. RESULTS: In this limited study, hypochloremia, hypokalemia, and metabolic alkalosis were present in 1 patient. One patient showed both hypochloremia and hypokalemia, whereas a third patient had only hypochloremia and a low normal serum potassium. The fourth patient demonstrated suspiciously low serum levels of Cl(-) and K(+). CONCLUSION: Because BN patients tend to be secretive regarding their disorder, serum electrolyte levels may serve as useful tools in diagnosing parotid swellings initiated by bulimia. PMID- 14561966 TI - The histologic differentiation of oral condyloma acuminatum from its mimics. AB - OBJECTIVE: The purpose of this study was to determine which histologic features could enable one to distinguish oral condyloma, as defined by the detection of human papillomavirus (HPV) DNA through in situ hybridization, from its mimics. STUDY DESIGN: Thirty-two paraffin-embedded specimens from 28 patients that were clinically suggestive of oral condyloma were analyzed histologically and through in situ hybridization with a consensus HPV probe. RESULTS: HPV DNA was detected in 17/32 (53%) of the lesions; no additional positive cases were detected after polymerase chain reaction amplification. Only 5 of the 17 virus-positive cases were considered to be histopathologically unequivocal for condyloma. The histologic features significantly associated with HPV detection were nonuniform perinuclear halos, often in association with epithelial crevices (P =.02), and papillomatosis (P =.02). Each of the 17 patients who were HPV-positive had either HPV-6 or HPV-11, a finding that is similar to those for condyloma involving the penis and vulva/vagina. CONCLUSION: Differentiation between oral condyloma and its mimics is best accomplished by using a combination of histologic, clinical, and in situ viral studies. PMID- 14561967 TI - Epithelioid blue nevus of the oral mucosa: a rare histologic variant. AB - Epithelioid blue nevus (EBN) is an extremely rare histologic variant of blue nevus that has only recently been identified. Unlike other variants of blue nevus, which primarily are composed of pigmented, spindle-shaped melanocytes, EBN is characterized by large, well-defined, heavily-pigmented polygonal or epithelioid-shaped melanocytes intermixed with less densely pigmented epithelioid and fusiform-shaped melanocytes. Furthermore, in contrast to other benign melanocytic proliferations, the lesional cells in EBN exhibit little or no maturation as they extend deeper into the underlying tissue. Blue nevi are the second most common form of nevus in the oral cavity. However, to our knowledge, the epithelioid variant has not been previously identified in the mouth. Only 6 examples of EBN have been identified in the skin of the head and neck. We now report the first documented case of EBN involving the oral mucosa. A brief review of the clinical and histopathologic features of EBN is also presented. PMID- 14561968 TI - Gingivostomatitis after eating fish parasitized by Anisakis simplex: a case report. AB - Anisakis simplex (AS) is a nematode that may be encountered as a parasite in various kinds of seafood. Human beings may accidentally acquire AS larvae by eating raw or undercooked seafood. In addition to human parasitization (anisakiasis), this nematode can induce allergic reactions. AS-related diseases are frequent, especially in those countries with a high level of fish consumption and with traditions of eating raw or undercooked seafood. To our knowledge, this is the first report of gingivostomatitis secondary to the ingestion of fish with AS parasites. PMID- 14561969 TI - Endodontic measurement accuracy and perceived radiograph quality: effects of film speed and density. AB - OBJECTIVES: This study sought to determine the effects of direct exposure x-ray film speed and background density on observer assessment of endodontic working lengths and on perceived radiographic image quality. STUDY DESIGN: A human cadaver maxilla section with surrounding soft tissues was used for the study. The canal length to the radiographic apex was determined on 4 canals in maxillary posterior teeth by using Trophy RVG images and adjusting the position of a No. 15 file in each canal until the file tip coincided with the radiographic apex in images made at 3 different vertical angulations. The files were measured with a micrometer from the file stop to the file tip to obtain the length to the radiographic apex. Then No. 10 files were placed in the 4 canals at varying lengths short of this previously determined length, and 5 observers assessed the distance from the file tip to the radiographic apex on radiographs made with Kodak D-, E-, and F-speed and Flow D- and E-speed direct exposure x-ray films that were exposed to produce background densities of 1.5, 2.0, and 3.0. Subjective appraisal of radiographic quality was also assessed. RESULTS: Analysis of variance and Tukey honestly significantly different post-hoc analysis results concerning measurement errors made with each film type revealed significantly less error for Kodak Ektaspeed Plus (E-speed) intraoral x-ray film than for Kodak InSight (F-speed) and Flow E; however, no difference was detected among Kodak Ektaspeed Plus (E-speed), Kodak Ultra-Speed (D-speed), and Flow D. Films with a background optical density of 3.0 received 98% favorable ratings; radiographs with a background optical density of 2.0 received 77% favorable ratings; and those with background optical density of 1.5 received only 18% favorable ratings at the 95% confidence level. Flow D film received the most favorable ratings, but there was no statistically significant difference among other film types at the 95% confidence level. CONCLUSIONS: Underexposed radiographs are perceived as inferior to slightly overexposed radiographs for endodontic file length assessment regardless of the film speed used. Current Flow and Kodak E-speed and F-speed radiographs appear to be as accurate as other accepted radiographs used in determining endodontic working lengths. Image background density should be kept constant when making comparisons among x-ray films. PMID- 14561970 TI - Periradicular radiographic assessment in diabetic and control individuals. AB - OBJECTIVE: The purpose of this study was to investigate the prevalence of radiographic periradicular radiolucencies in endodontically treated and untreated teeth in patients with and without diabetes. STUDY DESIGN: The records of 30 subjects with diabetes and 23 control subjects attending the Endodontic Graduate Clinic at the University of Florida, Gainesville, were reviewed. The number of teeth with root canal treatments with and without periradicular radiolucencies and the number of teeth without endodontic treatment but with periradicular lesions were recorded. RESULTS: There were no main effects of sex, diabetes diagnosis, or age (the covariate) on the 3 outcomes of interest- nonsurgical endodontic treatment (NSE) with lesions, NSE without lesions, and no NSE with lesions. However, there were significant interactions between sex and diabetes diagnosis for both of the endodontic outcomes, NSE with lesions (F = 4.292; P <.05) and NSE without lesions (F = 4.241; P <.05). This meant that men with type 2 diabetes who had endodontic treatments were more likely to have residual lesions after treatment. CONCLUSION: Type 2 diabetes is associated with an increased risk of ill response by the periradicular tissues to odontogenic pathogens. PMID- 14561971 TI - Effect of different temperatures and penetration depths of a System B plugger in the filling of artificially created oval canals. AB - OBJECTIVES: The purpose of this study was to compare the gutta-percha-filled area in oval canals while varying the temperature and penetration depth of the System B plugger. STUDY DESIGN: Seventy-five resin blocks, with artificially created oval canals, were obturated by using a continuous wave technique. In groups 1, 2, and 3, the plugger was introduced to a depth of 4 mm from the apex at set temperatures of 100, 200, and 350 degrees C, respectively. In group 4, the plugger was inserted to a depth of 2 mm from the apex at a set temperature of 200 degrees C. Group 5 was obturated in the same way as group 2, and then an additional cold plugging was performed to a depth of 3 mm from the apex. The cross sections of the obturated canals were photographed, and the percentage of the gutta-percha-filled area (PGP) was measured. RESULTS: No significant differences were found between the 3 different temperature settings (P >.05). At 1 mm from the apex, group 4 showed the highest PGP (95.57% +/- 6.86), followed by group 5 (84.72% +/- 12.19), and group 2 (67.33 +/- 13.03) (P <.05). CONCLUSION: In artificially created oval canals, plugger temperature within the range used in this study did not affect the PGP. However, the penetration depth of the plugger was significantly related to PGP. PMID- 14561972 TI - Biocompatibility of an experimental glass-ionomer cement sealer in rat mandibular bone. AB - OBJECTIVE: The purpose of this study was to compare an experimental glass-ionomer cement sealer, KT-308, with a conventional zinc oxide-eugenol sealer, Canals, in terms of tissue compatibility and solubility. STUDY DESIGN: Tissue reactions were examined under light and electron microscopes at 3 and 20 days after the implantation of either freshly mixed KT-308 sealer or Canals sealer into prepared cavities in rat mandibles. RESULTS: At 3 days after implantation, no inflammatory reaction was seen around KT-308 sealer, which was in direct contact with the bone surface. In contrast, Canals sealer elicited an initially severe inflammation in the surrounding tissue. At 20 days, the majority of KT-308 sealer remained in the bone cavity. Canals sealer was largely absorbed and surrounded by fibrous tissue with many macrophages. An ultrastructural examination also revealed that no intervening tissue was present between the cut bone surface and the glass-ionomer cement sealer and that disintegrated zinc oxide-eugenol particles were engulfed by macrophages. CONCLUSIONS: KT-308 sealer possesses better tissue compatibility and lower solubility compared with a conventional zinc oxide-eugenol sealer, suggesting its potential for use as a root canal sealant. PMID- 14561973 TI - An in vitro study of the torsional properties of new and used rotary nickel titanium files in plastic blocks. AB - OBJECTIVE: The purpose of this study was to compare torque (in grams per centimeter) and angle of rotation (in degrees) at the time of fracture of new and used nickel-titanium .04 ProFile rotary files. The relationship between the size of the instrument and the torque at fracture was also investigated. STUDY DESIGN: The torque and angle of rotation at fracture of new and used nickel-titanium .04 ProFile rotary files Nos. 15 to 40 were determined according to American National Standards Institute/American Dental Association Specification No. 28. Analysis of variance was used to compare the torque and angle of rotation at fracture of the different sizes of the new instruments and of new and used instruments of the same size (alpha =.05). The relationship between torque at fracture and the size of the instrument was determined by means of regression analysis. RESULTS: The torque at fracture of the new instruments increased significantly with the diameter. The used instruments, Nos. 25 to 40, had significantly lower values of torque at fracture than did the new ones. There were no statistically significant differences between the angle of rotation at fracture of the new and the used instruments (P >.05). A stronger relationship was found between the size of the file and the torque at fracture for the new instruments (P <.0001) than for the used ones (P <.0001). CONCLUSION: The results of the present study suggest that the values of the torque at fracture of new instruments increased significantly with the diameter. The results also suggest that repeated use of ProFile .04 instruments mainly affected the torque at fracture. PMID- 14561974 TI - A retrospective radiographic study of root-end surgery with amalgam and intermediate restorative material. AB - OBJECTIVE: We sought to compare amalgam with intermediate restorative material in terms of their effectiveness as root-end filling materials during root-end surgeries. STUDY DESIGN: A root-end operation was performed on 228 patients with 262 endodontically treated teeth with periapical lesions. Of these teeth, 122 from 101 patients were available for follow-up. Radiographs were taken immediately postoperatively and again at a mean of 11.25 months (range, 6-45 months) postoperatively. The radiographs were digitized, and the diameter of the periapical lesion was measured by means of a computerized system. The mesiodistal distance at the cementoenamel junction served as an internal standard. Radiographic findings were classified into 1 of 3 groups: complete healing, incomplete healing, or unsatisfactory (no) healing. RESULTS: Complete healing was observed in 44.3% of the teeth, incomplete healing in 21.3%, and unsatisfactory healing in 34.4%. No statistical significance was found for amalgam versus intermediate restorative material, but teeth that underwent additional operations had a significantly worse success rate than those that underwent only first-time operations. No correlation was found between the subjective prognosis by the surgeons and the long-term outcome. CONCLUSION: As root-end filling materials, amalgam and intermediate restorative material had the same clinical effectiveness. The classic root-end operation is unpredictable. Reoperations should be reconsidered and avoided if possible. PMID- 14561975 TI - Substance P induces interleukin-8 secretion from human dental pulp cells. AB - OBJECTIVE: The role of neuropeptides in recruiting leukocytes in dental pulps is not known; therefore, we investigated whether interleukin-8 (IL-8) secretion from human pulp cells is increased after stimulation with substance P (SP) or calcitonin gene-related peptide (CGRP). METHODS: Primary pulp cells exhibiting a fibroblast-like phenotype and an endothelial cell line were stimulated with various doses of SP or CGRP, and IL-8 secretion was measured by enzyme-linked immunosorbent assays. RESULTS: IL-8 secretion from pulp cells increased significantly at 10(-8) to 10(-4) mol/L of SP stimulation (5- to 13.8-fold; P <.05); however, no significant IL-8 increase with CGRP (up to 10(-4) mol/L) stimulation was observed, nor was there synergistic induction of IL-8 with SP plus CGRP. The IL-8 increase reached its maximum at 8 hours after SP stimulation of the pulp cells. SP at a concentration of 10(-4) mol/L created minimal IL-8 induction in the endothelial cells and no synergistic induction by SP and CGRP. CONCLUSIONS: Pulp cells may up-regulate IL-8 secretion by SP stimulation, which suggests that SP released in dental pulp may play a role in the influx of leukocytes, attracted by IL-8, into the pulp tissue. PMID- 14561976 TI - Perceived quality of radiographic images after rapid processing of D- and F-speed direct-exposure intraoral x-ray films. AB - OBJECTIVE: We sought to compare the densitometric properties and perceived image quality of InSight (F-speed) and Ultra-Speed (D-speed) film radiographs processed with rapid chemistry. The effects of density, contrast, and film speed on perceived image quality were also studied. STUDY DESIGN: Images were made of a human cadaver phantom with exposures to achieve background densities approximating 1.5, 2.0, and 3.0. Films were processed in a radiographic darkroom by using Insta-Neg and Insta-Fix rapid chemistry as the manufacturer had recommended. Five endodontic residents independently analyzed images of varying density, speed, and contrast that were presented in a randomized manner. They were required to evaluate the perceived image quality of 5 specifically designated areas on the film, using a labeled photograph as a guide. These areas included root canal obturation, periodontal ligament space, dentinoenamel junction, and crestal bone height. In addition, they were also asked to assess the overall perceived image quality. Statistical analysis consisted of ordinal regression and 2-factor analysis of variance. RESULTS: No statistically significant differences were proved between F- and D-speed radiographs within the same density group. Higher density and higher contrast resulted in a statistically significant positive impact (P <.01) on the ranking for all 5 subjective determinations. Observers preferred the films exposed to a background density of 3.0 over those of a lower density (P <.01). CONCLUSIONS: InSight (F speed film) can be used with rapid chemistry to ensure less radiation exposure to patients than is necessary with D-speed film. The observers participating in this study preferred radiographs from the 3.0 background density group to those from the 2.0 and 1.5 density groups. PMID- 14561977 TI - Cytokines in periradicular lesions: the effect of linezolid treatment. AB - OBJECTIVE: The purpose of this study was to quantify the cytokines interleukin (IL)-1 receptor antagonist (RA), IL-6, and transforming growth factor beta(1) found in extracts of inflammatory periapical tissues and to study the effect of a new antibiotic (linezolid) on the levels of these cytokines. STUDY DESIGN: Twenty two patients with root-filled teeth with persisting periapical pathoses were randomly divided into a control group or an antibiotic group. One tooth from each of the patients was resected at the root-end, and the periapical tissue was collected. IL-1RA, IL-6, and TGF-beta(1) were quantified in tissue extracts through the use of enzyme-linked immunosorbent assays. RESULTS: Measurable amounts of all 3 cytokines were found in all extracts. A statistically significant reduction in IL-1RA per milliliter was observed in the linezolid group in comparison with the control group (P <.05). The difference in variation of IL-1RA in the treatment and control groups was also highly statistically significant (P <.001). CONCLUSIONS: Although drawn from a limited sample size, the results indicate that IL-1RA is a sensitive indicator of the effect of antibacterial treatment on the immune response in periapical inflammatory tissues. PMID- 14561978 TI - Magnetic resonance imaging of normal and osteomyelitis in the mandible: assessment of short inversion time inversion recovery sequence. AB - OBJECTIVE: We sought to determine the suitable magnetic resonance imaging conditions for the short inversion time inversion recovery (STIR) sequence through the use of phantoms; to describe the signal characteristics of normal structures in the mandible; and to evaluate the usefulness of STIR images in enabling the identification of mandibular osteomyelitis on conventional T1- and T2-weighted spin-echo images. STUDY DESIGN: Suitable mandibular STIR imaging conditions were determined by varying inversion time and repetition time in each sequence. STIR magnetic resonance images of 162 healthy subjects and STIR and T1- and T2-weighted spin-echo images of 21 subjects with mandibular osteomyelitis were evaluated. RESULTS: In STIR imaging, the signal of oil was suppressed at an inversion time equaling 100 milliseconds and a repetition time equaling 1500 to 3000 milliseconds. In healthy subjects, the mandibular marrow was revealed to have high signal intensities (100%) and cortical bone had no signal intensities (100%) on STIR images. In surrounding soft tissue in these healthy subjects, the submandibular glands were shown to have high signal intensities (100%); the parotid glands had intermediate to high signal intensities (100%); the sublingual glands had high (88.9%) and intermediate to high (11.1%) signal intensities; lymph nodes had high signal intensities (100%); and the masseter muscles had intermediate signal intensities (100%) on STIR images. The lesions in bone marrow had low (75%) and low to intermediate (25%) signal intensities on T1-weighted images and high (54%), intermediate to high (29%), and intermediate (17%) signal intensities on T2-weighted images. On STIR images, the signal intensities resulted in high (75%), intermediate to high (21%), and intermediate (4%) signal intensities. CONCLUSIONS: STIR imaging is highly effective for the evaluation of bone marrow and surrounding soft tissue in terms of the detection of osteomyelitis in the mandible and the identification of inflammation spreading to soft tissue. PMID- 14561979 TI - Radiation absorbed in maxillofacial imaging with a new dental computed tomography device. AB - OBJECTIVE: The purpose of this investigation was to measure the tissue-absorbed dose and to calculate the effective dose for the NewTom 9000, a new generation of computed tomographic devices designed specifically for dental applications. Comparisons are made with existing reports on dose measurement and effective dose estimates for panoramic examinations and other computed tomographic imaging modalities for dental implants. STUDY DESIGN: Thermoluminescent dosimeters were implanted in a tissue-equivalent humanoid phantom at anatomic sites of interest. Absorbed dose measurements were obtained after single and double exposures. The averaged tissue-absorbed doses were used for the calculation of the whole-body effective dose. RESULTS: The effective dose for imaging of maxillomandibular volume with a NewTom 9000 machine is 50.3 muSv. CONCLUSION: The effective dose with the NewTom 9000 machine is significantly less than that achieved with other computed tomographic imaging methods and is within the range of traditional dental imaging modalities. PMID- 14561984 TI - Are predictive factors clinically useful in bone marrow transplantation? AB - Variables that accurately predict the clinical outcome of any procedure, including bone marrow transplantation (BMT), are of paramount importance when assessing the risks and benefits of the procedure. This review of the world's literature of variables affecting overall outcome after myeloablative BMT critically appraises the value of many bone marrow transplant dogmas. There is a relative paucity of data supporting many commonly used transplant practices, including having an upper age limit for eligibility criteria, and absolute requirements for cardiac and pulmonary function pre transplant. In contrast, recently published literature suggests that several parameters occurring soon after a BMT has been performed may accurately predict transplant outcome. Ultimately, given the rapidly evolving nature of BMT, renewed clinical research of variables predictive of transplant outcome is needed. PMID- 14561985 TI - Localized radiation increases morbidity and mortality after TBI-containing autologous stem cell transplantation in patients with lymphoma. AB - The purpose of this study is to assess the relationship between involved field radiation therapy (IFRT) and treatment-related morbidity and mortality in patients receiving high-dose chemotherapy (HDC), total body irradiation (TBI) and autologous peripheral stem cell transplant (PSCT) for Hodgkin's and non-Hodgkin's lymphoma. Between January 1994 and May 2002, 156 patients underwent HDC, TBI and autologous PSCT. Localized external beam radiation therapy was given to 21 patients for consolidation, or to achieve control of symptomatic or active disease prior to or after transplant. Among patients who had IFRT prior to autologous PSCT, five treatment-related deaths were observed, compared to seven deaths in 135 patients who had autologous PSCT without IFRT (P<0.01). Most deaths were attributable to sepsis and multiorgan failure. A higher incidence of pneumonitis was also noted in patients exposed to mediastinal irradiation. No adverse impact on long-term survival could be demonstrated. Involved field radiation prior to TBI is associated with higher treatment-related mortality in lymphoma patients undergoing autologous peripheral stem cell transplant, necessitating careful monitoring. PMID- 14561986 TI - Allogeneic stem cell transplantation with reduced-intensity conditioning is potentially feasible as an outpatient procedure. AB - Allogeneic stem cell transplantation (allo-SCT) after a reduced-intensity conditioning (RIC) protocol is associated with decreased short-term toxicity. This suggests that the procedure could be performed on an outpatient basis. We analysed the incidence and risk factors of grade >or=2 conditioning-related toxicities (CRTs) as a hallmark for hospital admission, in 41 consecutive patients allografted from an HLA identical sibling after RIC. The RIC regimen consisted of fludarabine plus melphalan for lymphoid malignancies, and fludarabine plus busulphan for myeloid malignancies. In all, 11 patients (27%) did not experience any toxicity. The more frequent CRTs observed were neutropenic fever and gastrointestinal toxicity. The median duration of hospitalisation was 27 (range, 17-50) days. If allo-SCT had been planned as an outpatient procedure and admission indicated only in the case of >or=2 CRTs, the inpatient period would have decreased to 9 (range, 0-33) days (P<0.001). No risk factors for CRTs were identified. Allo-SCT after an RIC regimen is a well-tolerated procedure. Our results warrant a prospective pilot trial of nonmyeloablative allo-SCT performed in the outpatient setting. PMID- 14561987 TI - Differences between graft product and donor side effects following bone marrow or stem cell donation. AB - We report graft product stem cell yields and donor safety results of a randomized multicenter study comparing allogeneic peripheral blood stem cell (PBSC) PBSC transplantation with BM transplantation. Matched HLA-identical sibling donors (n=329) were randomized to filgrastim-mobilized PBSC or bone marrow (BM) donation groups. Median yields per kg recipient weight of CD34(+) cells, T cells, and natural killer (NK) cells, respectively, were approximately two-fold, eight-fold, and greater than eight-fold in the PBSC group than in the BM group (CD34(+) cells, 5.8 x 10(6)/kg vs 2.7 x 10(6)/kg; T cells, 300.1 x 10(6)/kg vs 35.7 x 10(6)/kg; NK cells, 28.2 x 10(6)/kg vs 3.6 x 10(6)/kg; P<0.001 for each). In connection with the cell collection procedures, PBSC donors spent a shorter median time in hospital than BM donors (0 vs 2 days; median difference -2 days, 95% CI -2 to 2) and had fewer median days of restricted activity (2 vs 6 days; median difference -3 days, 95% CI -4 to 2). Overall, 65% of PBSC donors and 57% of BM donors reported at least one adverse event (AE), most of which were transient, mild-moderate in severity, and without clinical sequelae. PBSC donors experienced predominantly filgrastim-related AEs, while BM donors experienced predominantly harvest-related AEs. PMID- 14561988 TI - Clinical and pharmacological risk factors for acute graft-versus-host disease after paediatric bone marrow transplantation from matched-sibling or unrelated donors. AB - The aim of this study was to identify the risk factors for acute graft-versus host disease (aGVHD) in children transplanted from a matched-sibling donor (MSD) or an unrelated donor (UD). In all, 87 children consecutively underwent allogeneic bone marrow transplantation (BMT) from MSD (n=36), and UD (n=51). GVHD prophylaxis included CsA alone (n=33) or with MTX (n=51). ATG was added in UD-BMT and thalassemic recipients. CsA whole-blood concentrations were measured by EMIT and the dosing regimen was monitored by Bayesian pharmacokinetic modelling. Trough blood concentration (TBC) during the first 2 weeks post transplantation was lower in children who developed grade II-IV aGVHD than those developing no GVHD or only grade I (57+/-9 vs 94+/-8 ng/ml, P=0.007), whereas peak blood concentration and area under concentration curve vs time were similar in both groups. TBC <85 ng/ml and 'use of MTX' were associated with aGVHD in MSD-SCT (P=0.003 and 0.007, respectively) as well as in UD-SCT (P=0.006 and 0.003). Donor age >or=8 years was significant only in MSD-BMT. Our results have shown the significant decisive role of pharmacological factors such as CSA TBC or use of MTX in the occurrence of GVHD in MSD as well as in UD paediatric BMT. PMID- 14561990 TI - The graft-versus-leukemia effect of nonmyeloablative stem cell allografts may not be sufficient to cure chronic myelogenous leukemia. AB - We treated 12 patients with chronic myelogenous leukemia (CML) with a low intensity preparative regimen followed by allogeneic stem cell transplantation in an attempt to confer a curative graft-versus-leukemia (GVL) effect with minimum morbidity. Seven patients in first chronic phase (CP1) and five in second chronic phase (CP2) (age 15-68 years) received a nonmyeloablative conditioning regimen of fludarabine and cyclophosphamide, followed by a G-CSF-mobilized peripheral blood stem cell (PBSC) transplant from an HLA-identical sibling. Cyclosporine (CsA) was used for graft-versus-host disease (GVHD) prophylaxis. Median follow-up was 384 days. Neutrophil recovery occurred at a median of 12 days. There was no transplant-related mortality. Of the seven CP1 patients transplanted, seven achieved a stable molecular remission; two with no post-transplant intervention, three after donor lymphocytes, imatinib and interferon, and two after a myeloablative stem cell transplant. Four of five CP2 patients died in blast crisis and one survived in molecular remission. Of the 12 patients with durable engraftment, six had Grades II-IV acute GVHD; six had limited chronic GVHD. These results suggest that cytoreduction is required to optimize the curative effect of allogeneic stem cell transplantation for CML. PMID- 14561989 TI - Autologous peripheral blood stem cell transplantation with induction of autologous graft-versus-host disease in acute myeloid leukemia. AB - We studied whether the induction of autologous graft-versus-host disease (GVHD) has an antileukemic effect and consequently increases the survival of patients undergoing autologous peripheral blood stem cell transplantation (PBSCT). In all, 22 acute myeloid leukemia patients with favorable and intermediate cytogenetic risk, in their first complete remission, were administered cyclosporine c.i.v. from day 0 to day +28 at a dose of 3.0 mg/kg per day and interferon-gamma (IFN gamma) at 0.025 mg/m(2) s.c. every other day from day +14 to day +42 following autologous PBSCT. Natural-killer (NK)-cell activity assays and skin biopsies were performed. Successful engraftment was achieved in all patients at a median of 13 days without significant additional toxicity. Histologically confirmed cutaneous GVHD developed in 12 patients, and NK-cell activity was significantly augmented after autologous PBSCT in those patients (P=0.03). After a median follow-up duration of 37.7 months (range, 7.3-72.8), the 3-year disease-free survival (DFS) and overall survival (OS) rates were 64.4 and 73.1%, respectively, without significant correlation with GVHD status or augmentation of NK-cell activity. These data suggest that the administration of cyclosporine and IFN-gamma following autologous PBSCT improves OS and DFS, which may be attributable to the antileukemic effect, although no difference in survival could be demonstrated between cutaneous GVHD-positive and -negative groups. PMID- 14561991 TI - Multivariate analysis of risk factors for hemorrhagic cystitis after hematopoietic stem cell transplantation. AB - To establish the most appropriate prophylactic therapy and risk factors for predicting hemorrhagic cystitis (HC) after stem cell transplantation (SCT), we retrospectively analyzed the clinical records of 450 transplant patients treated from 1982 to 2002. In all, 81 patients developed early- and/or late-onset HC (early=29, late=48, both=4). For the incidence of early-onset HC, administration of cyclophosphamide (CY) (p=0.0079, odds ratio (OD)=5.109, 95% confidence interval (CI)=1.533-17.030), busulfan (BU) (p=0.0015, OD=3.336, 95% CI=1.584 7.027), BU+CY (p=0.0001, OD=4.369, 95% CI=2.055-9.292), antithymocyte globulin (p=0.0009, OD=3.368, 95% CI=1.642-6.911), nonradiation (p=0.0163, OD=2.564, 95% CI=0.181-0.841), 2-mercaptoethane sodium sulfonate (Mesna) (p=0.0001, OD=7.519, 95% CI=2.847-19.858), and bladder irrigation (p=0.0001, OD=4.950, 95% CI=2.328 10.523) were risk factors. By Fisher's exact test, the combination of BU and Mesna was a more significant risk factor (P<0.001) than Mesna alone (p=0.008) compared to the administration of neither agent. By multivariate analysis, prophylactic administration of Mesna (p=0.0105, OD=5.301, 95% CI=1.477-19.026) and bladder irrigation (p=0.0001, OD=9.469, 95% CI=3.872-23.156) were significant risk factors of early-onset HC. We conclude that (i). high-dose BU as well as CY is a cause of HC, (ii). protective bladder irrigation has an opposite effect, and (iii). Mesna possibly has a toxic effect on bladder mucosa. PMID- 14561992 TI - Autografting as a risk factor for persisting iron overload in long-term survivors of acute myeloid leukaemia. AB - We studied the iron status of 32 evaluable adult acute myeloid leukaemia (AML) survivors who were entered into the UK Medical Research Council acute myeloid leukaemia (AML) 10 and 12 trials at our institution between 1988 and 1998. Patients were required to have been independent of all blood products for at least 3 years. As a group, the median first serum ferritin level was 1323 mug/l (NR 19-300 mug/l) at a median of 1321 days from the last transfusion confirming the presence of significant iron overload persisting for some years after completion of all therapy and blood products. There was a general trend for the serum ferritin level to fall with time, but the fall was less pronounced in men and carriers of the C282Y mutation. Recipients of autologous stem cell transplantation (SCT) had a higher median first serum ferritin level (3245 mug/l) than patients who received chemotherapy alone (1148 mug/l) or allogeneic SCT (1334 mug/l) because of increased use of transfused blood. Nine of the 10 recipients of autologous SCT underwent venesection. No evidence of end organ damage was seen in any patient. Serial monitoring of serum ferritin and assessment of the C282Y status may be useful in all long-term AML survivors, especially autograft recipients. PMID- 14561993 TI - Second malignancy following high-dose therapy and autologous stem cell transplantation: incidence and risk factor analysis. AB - To establish incidence and risk factors for development of second malignant neoplasms after high-dose chemo/radiotherapy (HDT) and autologous hematopoietic stem cell transplantation (AHSCT), the case files of 800 consecutive patients who underwent AHSCT at our institution between June 1982 and December 2000 were reviewed. In all, 26 patients developed 29 second malignancies (nine myelodysplastic syndrome (MDS)/acute myelogenous leukemia (AML), 16 solid tumors and four lymphoproliferative disorders (LPDs)) for a 15-year cumulative incidence of 11% (95% confidence interval (CI), 5-18%). These second tumors occurred at a median of 68 (range 1.5-177) months following AHSCT. The relative risk (RR) compared to the general population of developing a second malignancy following AHSCT was 3.3 (CI 2.2-4.7) P<0.001. The RR of developing MDS/AML, LPD and a solid tumor was 47.2 (CI 21.5-89.5) P<0.001, 8.1 (2.2-20.7) P=0.002 and 1.98 (1.1-3.2) P=0.009, respectively. In multivariate analysis, age >or=35 years at the time of AHSCT (P=0.001) and an interval from diagnosis to AHSCT >or=36 months (P=0.03) were associated with a greater risk of developing a second malignancy. Patients who have undergone HDT and AHSCT are at significant risk for developing a second malignancy and should receive indefinite follow-up. PMID- 14561994 TI - Incidence and treatment of hemorrhagic cystitis in children given hematopoietic stem cell transplantation: a survey from the Italian association of pediatric hematology oncology-bone marrow transplantation group. AB - The purpose of this multicenter study was to assess the incidence and the treatment of hemorrhagic cystitis (HC) in 1218 pediatric patients, with a mean age of 10.8 years, who underwent hematopoietic stem cell transplantation (HSCT). In all, 44 patients (3.6%) developed HC a median 23 days after HSCT. The incidence of HC was higher in allogeneic than in autologous HSCT recipients (P=0.0001). Of the 44 patients, 37 (84%) recovered from HC in a median 30 days (range 3-100); the other seven children died while still suffering from HC. Hyperbaric oxygen therapy (HOT) achieved significantly better results than prostaglandin therapy (P=0.02) in the treatment of grade II-III HC. By multivariate analysis, age <96 months and allogeneic HSCT were significantly associated with the occurrence of HC: P=0.008 and 0.013, respectively. After a median follow-up of 5.75 years, the 5-year survival of patients who did or did not develop HC was: 43 vs 52%, P=0.03, respectively. This study indicates that age and type of HSCT are factors predisposing to HC in children given HSCT and demonstrates the promising role of HOT in a conservative approach to HC treatment. PMID- 14561995 TI - Sexual functions after high-dose chemotherapy in survivors of germ cell tumors. AB - We investigated the changes in sexual function in male patients with germ cell tumor continuously disease free after one or two courses of high-dose chemotherapy with hematopoietic stem cell support. A questionnaire was mailed to 35 patients, and 30 patients sent it back. Sexuality was considered a problem by 10 patients (33%), but no patients considered sexuality a major problem. Erection was more difficult to achieve in seven patients (23%) and 10 patients (33%) experienced increased difficulty in maintaining an erection. Eight patients (27%) had the experience of less intensive and less frequent orgasm. In all, 13 patients (43%) thought that both the disease and treatment had worsened their sexual capacity, but 20 patients (67%) were satisfied with their sex life. Most of the patients (63%) considered that insufficient information and counselling had been given by their physicians about the sexual sequelae of therapy. However, the amount of information about the disease and treatment was considered good by 77 and 80% of the patients, respectively. This study shows that 27% of patients were not content with their ability to attain sexual satisfaction due to the illness or its treatment. Communication is an important issue and better information tools could lead to improved compliance in these patients. PMID- 14561996 TI - Procalcitonin and C-reactive protein do not discriminate between febrile reaction to anti-T-lymphocyte antibodies and Gram-negative sepsis. AB - Treatment with antibodies against T-lymphocytes usually triggers a febrile response potentially mimicking or masking infection. Procalcitonin (PCT) is considered a sensitive and specific marker of systemic bacterial and fungal infection. It was the aim of this study to investigate the characteristics of PCT and C-reactive protein (CRP) during treatment with polyclonal or monoclonal anti T-cell antibodies, in order to examine the ability of these parameters to distinguish between systemic bacterial infection and reaction to antibody treatment. Thus, 15 consecutive febrile episodes after T-cell antibody infusion without clinical signs of infection were compared with nine episodes of Gram negative sepsis. After T-cell antibody infusion PCT and CRP serum levels increased to a similar extent as in Gram-negative sepsis. Therefore, during T cell antibody treatment neither PCT nor CRP are adequate for differentiating between fever due to infection or to unspecific cytokine release. PMID- 14561997 TI - Constitution and telomere dynamics of bone marrow stromal cells in patients undergoing allogeneic bone marrow transplantation. AB - We evaluated the genotypic origin of mesenchymal stem cells (MSC) following sex mismatched allogeneic bone marrow transplantation (BMT), and investigated the telomere dynamics in MSC in normal individuals and patients after BMT. The study population consisted of 11 patients with hematologic disorders who showed complete chimerism after BMT. Telomere length was measured in MSC using Southern blotting analysis in eight patients and 18 healthy subjects as a control group. Following culture, MSC were identified by the expression of SH2 and SH4, and lack of CD14, CD34, and CD45. All MSC showed the recipient genotype, based on the results of fluorescent in situ hybridization analysis using X-chromosome satellite probes or microsatellite DNA polymorphism analysis. The mean telomere length in MSC from normal controls was 7.2+/-0.53 kb (range, 6.12-7.78), and progressive telomere shortening was seen with age. There was no significant difference in MSC telomere length between the BMT group and age-matched controls. This study confirmed that the MSC isolated from the recipients of allogeneic BMT did not have the donor genotype, despite complete chimerism. Moreover, MSC were demonstrated to show progressive loss of telomere length with age, but the telomeres in MSC were not affected by BMT. PMID- 14561998 TI - Late occurrence of chronic immune-mediated axonal polyneuropathy following bone marrow transplant for juvenile-onset alpha-mannosidosis. AB - A 23-year-old woman with juvenile-onset alpha-mannosidosis developed an axonal polyneuropathy more than a year following successful unrelated donor (URD) BMT complicated by chronic graft-versus-host disease (GVHD). Progressive muscle weakness and paresthesias developed over at least 4 months, and made her nonambulatory. Nerve conduction and EMG studies demonstrated an axonal sensorimotor neuropathy. Cerebral spinal fluid (CSF) IgG was elevated with two peaks not identified in serum. Strength improved after a single course of plasma exchange and continued to improve over 12 months. The response to plasma exchange, elevated CSF IgG production, and evidence of a serum IgM peak suggest an immune-mediated mechanism. Chronic polyneuropathies following BMT are rare and are usually temporally related to GVHD or infection. This patient's disease was unusual because of its late occurrence and chronic onset in the face of resolved GVHD and in the absence of infection. PMID- 14561999 TI - Continued neurocognitive development and prevention of cardiopulmonary complications after successful BMT for I-cell disease: a long-term follow-up report. AB - I-cell disease or mucolipidosis type II, a rare inherited storage disorder of lysosomal enzyme localization, is characterized by dysostosis multiplex, progressive severe psychomotor retardation and death by 5-8 years from congestive heart failure and recurrent pulmonary infections. A 19-month old girl with I-cell disease received a bone marrow transplant (BMT) from an HLA-identical carrier brother. At the age of 7 years, 5 years after BMT, she has no history of respiratory infections. Her cardiac function remains normal with a shortening fraction of 47%, and she continues to gain neurodevelopmental milestones, albeit at a very slow rate. Musculoskeletal deformities have worsened despite BMT. This is the first report describing neurodevelopmental gains and prevention of cardiopulmonary complications in I-cell disease after BMT. PMID- 14562000 TI - Massive ascites of donor T-cell origin in a patient with acute GVHD after a reduced-intensity allograft for CLL. PMID- 14562001 TI - Clinical-grade cell purification from thawed cord blood: an example of translational research. PMID- 14562002 TI - Pertussis in adolescence after unrelated cord blood transplantation. PMID- 14562004 TI - Hydrogen ion dynamics and the Na+/H+ exchanger in cancer angiogenesis and antiangiogenesis. AB - Tumour angiogenesis and cellular pH regulation, mainly represented by Na(+)/H(+) antiporter exchange, have been heretofore considered unrelated subfields of cancer research. In this short review, the available experimental evidence relating these areas of modern cancer research is introduced. This perspective also helps to design a new approach that facilitates the opening and development of novel research lines oriented towards a rational incorporation of anticancer drugs into more selective and less toxic therapeutic protocols. The final aim of these efforts is to control cancer progression and dissemination through the control of tumour angiogenesis. Finally, different antiangiogenic drugs that can already be clinically used to this effect are briefly presented. PMID- 14562003 TI - Clinical use of rituximab in haematological malignancies. AB - Rituximab is a chimeric human/mouse monoclonal antibody that is approved for the treatment of relapsed and refractory non-Hodgkin's lymphoma (NHL) and in combination with CHOP (cyclophosphamide, doxorubicin, vincristine, prednisone) chemotherapy as first-line therapy for diffuse large B-cell NHL, where it has shown the first survival advantage over CHOP alone in more than 20 years. Strategies to help define the optimal therapeutic usage of rituximab are being assessed, including first-line and maintenance or extended therapy, and the combination of rituximab with chemotherapy in indolent NHL. Emerging data suggest that earlier use may yield higher response rates, extended therapy can prolong remission, and the addition of rituximab to chemotherapy can increase clinical and molecular remission rates when compared with those achieved using chemotherapy alone. Studies in the peritransplant setting suggest a role for rituximab in vivo purging prior to transplant and/or maintenance rituximab as a means of clearing minimal residual disease. Rituximab has also shown activity in other B-cell disorders such as chronic lymphocytic leukaemia. The full potential of this immunotherapeutic agent remains to be defined in ongoing and future clinical trials. PMID- 14562005 TI - Is it acceptable to approach colorectal cancer patients at diagnosis to discuss genetic testing? A pilot study. AB - In this pilot study, the acceptability of approaching 111 newly diagnosed colorectal cancer patients with the offer of genetic testing for hereditary nonpolyposis colorectal cancer (HNPCC) was assessed. A total of 78% of participants found it highly acceptable to have the information about HNPCC brought to their attention at that time. PMID- 14562006 TI - Imatinib administration in two patients with liver metastases from GIST and severe jaundice. AB - Imatinib is the only effective and approved systemic therapy for the treatment of patients with advanced gastrointestinal stromal tumours (GISTs). Although metastases from GISTs most commonly involve the liver, yielding hyperbilirubinaemia, very few data on imatinib administration in subjects with jaundice are available. We provide evidence that imatinib tolerability was not adversely affected by jaundice in two patients with advanced GIST. PMID- 14562007 TI - Industry-sponsored economic studies in oncology vs studies sponsored by nonprofit organisations. AB - The purpose of this analysis of health economic studies in the field of oncology was to investigate among sponsored studies whether any relationship could be established between the type of sponsorship and (1) type of economic analysis, (2) health technology assessed, (3) sensitivity analysis performed, (4) publication status, and (5) qualitative conclusions about costs. The Health Economic Evaluations Database (HEED, version 1995-2000) was searched on the basis of oncological ICD-9 codes, sponsorship, and comparative studies. This search yielded a total of 150 eligible articles. Their evaluations were prepared independently by two investigators, on the basis of specific criteria. When evaluators disagreed, a third investigator provided a deciding evaluation. There was no statistically significant relationship between the type of sponsorship and sensitivity analysis performed (P=0.29) or publication status (P=0.08). However, we found a significant relationship between the types of sponsorship and of economic analysis (P=0.004), the health technology assessed (P<0.0001), and qualitative cost assessment (P=0.002). Studies with industrial sponsorship were 2.56 (99% lower confidence interval (CI)=1.28) times more likely to involve cost minimisation analyses, were 0.04 (99% higher CI=0.39) times less likely to investigate diagnostic screening methods, and were 1.86 (99% lower CI=1.21) times more likely to reach positive qualitative conclusions about costs than studies supported by nonprofit organisations. In conclusion, our results suggest that there is a greater probability that industry-sponsored economic studies in the field of oncology tend to be cost-minimisation analyses, to investigate less likely diagnostic screening methods, and to draw positive qualitative conclusions about costs, as compared to studies supported by nonprofit organisations. PMID- 14562008 TI - A phase II trial with rosiglitazone in liposarcoma patients. AB - Agents of the thiazolidinedione drug family can terminally differentiate human liposarcoma cells in vitro by activating genes responsible for lipocyte differentiation. One study has shown clinical activity of troglitazone treatment in liposarcoma patients. We sought to find further evidence for this result. In all, 12 patients with a liposarcoma received rosiglitazone 4 mg b.d. They were followed clinically and with repeated biopsies for histological and biological studies. At the molecular level the mRNA translation of three genes that are induced by this treatment (peroxisome proliferator-activated receptor gamma (PPARgamma), adipsin and fatty acid binding protein) was determined. Nine patients were eligible for evaluation. One patient had to stop treatment due to hepatotoxicity. The mean time to progression was 6 months (2 - 16 months), with one patient still on treatment. We did not see any significant change in histologic appearance of the liposarcomas by the treatment. The level of gene expression changed significantly in two patients, but this did not result in a clinical response. Based on this study, rosiglitazone is not effective as an antitumoral drug in the treatment of liposarcomas. Increased PPARgamma activity does not correlate with the clinical evolution. PMID- 14562009 TI - Prognostic value of CA 19-9 levels in patients with inoperable adenocarcinoma of the pancreas treated with gemcitabine. AB - Serum carbohydrate antigen 19-9 (CA 19-9) has been identified as a useful tumour marker for diagnosis of exocrine pancreatic carcinoma, but its value for evaluating the response to chemotherapy with gemcitabine is not clear. Tumour regression in pancreatic carcinoma is hard to determine due to massive desmoplastic tissue. Furthermore, objective tumour response does not automatically transcribe into better survival. Therefore, clinical benefit response, a composed parameter consisting of factors like performance status, pain, and body weight was integrated in evaluating tumour response. The aim of this prospective study was to evaluate the usefulness of serial CA 19-9 measurements as a biochemical response marker and an outcome prognostic parameter in patients with advanced pancreatic cancer receiving gemcitabine treatment. A total of 46 consecutive patients (median age 66 years) suffering from histologically proven locally advanced or metastatic adenocarcinoma of the exocrine pancreas were analysed. Gemcitabine was applied for a median of 23 courses (range 6-76). Two patients achieved an objective complete remission, five an objective partial remission (overall response, OR=15.2%), while objective stable disease was documented in 19 and objective progressive disease in 20 patients. Patients with a decrease of >20% of the baseline CA 19-9 level after 8 weeks of chemotherapy had a significantly better median survival than patients with a rise or a decline <20%. The response of CA 19-9 >20% during chemotherapy was the only independent predictor of survival in a multivariate analyses. In contrast, neither objective tumour response nor clinical benefit response showed this level of significance. In conclusion, kinetics of CA19-9 serum concentration serves as an early indicator of response to gemcitabine chemotherapy in advanced pancreatic cancer. PMID- 14562011 TI - Postchemoembolisation syndrome--tumour necrosis or hepatocyte injury? AB - Transarterial chemoembolisation of liver tumours is typically followed by elevated body temperature and liver transaminase enzymes. This has often been considered to indicate successful embolisation. The present study questions whether this syndrome reflects damage to tumour cells or to the normal hepatic tissue. The responses to 256 embolisations undertaken in 145 patients subdivided into those with hepatocyte-derived (primary hepatocellular carcinoma) and nonhepatocyte-derived tumours (secondary metastases) were analysed to assess the relative effects of tumour necrosis and damage to normal hepatocytes in each group. Cytolysis, measured by elevated alanine aminotransferase, was detected in 85% of patients, and there was no difference in the abnormalities in liver function tests measured between the two groups. Furthermore, cytolysis was associated with a higher rate of postprocedure symptoms and side effects, and elevated temperature was associated with a worse survival on univariate analysis. Multivariate analysis demonstrated that there was no benefit in terms of survival from having elevated temperature or cytolysis following embolisation. Cytolysis after chemoembolisation is probably due to damage to normal hepatocytes. Temperature changes may reflect tumour necrosis or necrosis of the healthy tissue. There is no evidence that either a postchemoembolisation fever or cytolysis is associated with an enhanced tumour response or improved long-term survival in patients with primary or secondary liver cancer. PMID- 14562010 TI - A multicentre phase II trial of bryostatin-1 in patients with advanced renal cancer. AB - Protein kinase C (PKC) has a critical role in several signal transduction pathways, and is involved in renal cancer pathogenesis. Bryostatin-1 modulates PKC activity and has antitumour effects in preclinical studies. We conducted a multicentre phase II clinical trial in patients with advanced renal cancer to determine the response rate, immunomodulatory activity and toxicity of bryostatin 1 given as a continuous 24 h infusion weekly for 3 out of 4 weeks at a dose of 25 mug m(-2). In all, 16 patients were recruited (11 males and five females). The median age was 59 years (range 44-68). Patients had been treated previously with nephrectomy (8) and/or interferon therapy (9) and/or hormone therapy (4) and/or radiotherapy (6). Eight, five and three patients had performance statuses of 0, 1 and 2, respectively. A total of 181 infusions were administered with a median of 12 infusions per patient (range 1-29). Disease response was evaluable in 13 patients. Three patients achieved stable disease lasting for 10.5, 8 and 5.5 months, respectively. No complete responses or partial responses were seen. Myalgia, fatigue, nausea, headache, vomiting, anorexia, anaemia and lymphopenia were the commonly reported side effects. Assessment of biological activity of bryostatin-1 was carried out using the whole-blood cytokine release assay in six patients, two of whom had a rise in IL-6 levels 24 h after initiating bryostatin 1 therapy compared to pretreatment values. However, the IL-6 level was found to be significantly lower at day 28 compared to the pretreatment level in all six patients analysed. PMID- 14562012 TI - A phase II study of weekly cisplatin, 6S-stereoisomer leucovorin and fluorouracil as first-line chemotherapy for elderly patients with advanced gastric cancer. AB - The incidence of gastric cancer (GC) increases significantly after the fifth decade and palliative chemotherapy is the ultimate treatment in the majority of patients. We investigated safety and efficacy of a weekly regimen with cisplatin, fluorouracil and leucovorin as first-line chemotherapy for elderly patients with advanced GC. Chemotherapy-naive patients older than 65 years were considered eligible for study entry. Frail elderly patients were identified and excluded according to the following criteria: age >85 years, dependence in one or more activities of daily living (activities of daily living and instrumental activities of daily living scales), three or more comorbid conditions, one or more geriatric syndromes. Chemotherapy consisted of 1-day per week administration of intravenous cisplatin 35 mg m(-2), 6S-stereoisomer leucovorin 250 mg m(-2) and fluorouracil 500 mg m(-2) (PLF). Patients were re-evaluated after eight weekly cycles and six additional weekly administrations were planned for patients without disease progression. A 5-day subcutaneous filgrastim (5 mug Kg(-1) day( 1), days +1-+5) was used after the first treatment delay for neutropenia and maintained thereafter. In the whole group, the best intention-to-treat overall response rate was 43% (95% CI: 30-56%). The time to disease progression and the median survival time were 5.3 and 8.6 months, respectively. Fatigue was the commonest nonhaematologic toxicity (71% of the patients). Filgrastim was used in 30 patients who showed grade II (20 patients) or grade III (10 patients) neutropenia. Neither grade IV toxicity nor toxic deaths were observed. The weekly PLF regimen resulted safe and effective in elderly patients with advanced GC. This outpatient regimen is based on old and low-cost drugs and it may represent an alternative to new and more expensive combinations. PMID- 14562013 TI - Adjuvant and neoadjuvant therapy for gastric cancer using epirubicin/cisplatin/5 fluorouracil (ECF) and alternative regimens before and after chemoradiation. AB - Chemoradiation is now used more commonly for gastric cancer following publication of the US Intergroup trial results that demonstrate an advantage to adjuvant postoperative chemoradiotherapy. However, there remain concerns regarding the toxicity of this treatment, the optimal chemotherapy regimen and the optimal method of radiotherapy delivery. In this prospective study, we evaluated the toxicity and feasibility of an alternative chemoradiation regimen to that used in the Intergroup trial. A total of 26 patients with adenocarcinoma of the stomach were treated with 3D-conformal radiation therapy to a dose of 45 Gy in 25 fractions with concurrent continuous infusional 5-fluorouracil (5-FU). The majority of patients received epirubicin, cisplatin and 5-FU (ECF) as the systemic component given before and after concurrent chemoradiation. The overall rates of observed grade 3 and 4 toxicities were 38 and 15%, respectively. GIT grade 3 toxicity was observed in 19% of patients, while haematologic grade 3 and 4 toxicities were observed in 23%. Our results suggest that this adjuvant regimen can be delivered safely and with acceptable toxicity. This regimen forms the basis of several new studies being developed for postoperative adjuvant therapy of gastric cancer. PMID- 14562014 TI - Oxaliplatin- or irinotecan-based chemotherapy for metastatic colorectal cancer in the elderly. AB - The tolerance and efficacy of oxaliplatin and irinotecan for metastatic colorectal cancer are unknown in elderly patients. Methods. All consecutive patients over 74 years treated with oxaliplatin or irinotecan for metastatic colorectal cancer were enrolled. The tumour response was assessed every 2-3 months and toxicity was collected at each cycle according to World Health Organisation criteria. A total of 66 patients were enrolled from 12 centres. The median age was 78 years (range, 75-88 years); 39 patients had no severe comorbidity according to the Charlson score. In total, 44 and 22 patients received oxaliplatin or irinotecan, respectively, in combination with 5 fluororuracil+/-folinic acid or raltitrexed in 64 patients. A total of 545 chemotherapy cycles were administered in first (41%), second (51%) or third line (8%). A dose reduction occurred in 190 cycles (35%). Complete response, partial response and stabilisation occurred in 1.5, 20 and 47% of patients, respectively. The median time to progression and overall survival were 6.8 and 11.2 months in first line and 6.3 and 11.6 months in second line, respectively. Grade 3 and 4 toxicity occurred in 42% of patients: neutropenia 17%, diarrhoea 15%, neuropathy 11%, nausea and vomiting 8% and thrombopenia 6%. There was no treatment-related death. In selected elderly patients, chemotherapy with oxaliplatin or irinotecan is feasible with manageable toxicity. PMID- 14562015 TI - Enduring impact of communication skills training: results of a 12-month follow up. AB - The efficacy of a communication skills training programme was shown through a randomised trial. Oncologists (N=160) from 34 cancer centres were allocated to written feedback plus course; course alone; written feedback alone or control. Each clinician had 6 - 10 interviews with patients videotaped at baseline and 3 months postintervention. Analysis of videotapes revealed improvements in the communication skills of clinicians randomised to training (n=80) compared with others (n=80). A 12-month follow-up assessment is reported here. Robust Poisson conditional analyses of counts of changes in communication behaviours revealed no demonstrable attrition in those who had shown improvement previously, including fewer leading questions, appropriate use of focused and open-ended questions and responses to patient cues. Additional skills, not apparent at 3 months, were now evident; the estimated effect sizes corresponded to 81% fewer interruptions (P=0.001) and increased summarising of information to 38% (P=0.038). However, expressions of empathy (54%, P=0.001) declined. The overall results show that 12 15 months postintervention, clinicians had integrated key communication skills into clinical practice and were applying others. This is the first RCT to show an enduring effect of communication skills training with transfer into the clinic. PMID- 14562016 TI - Cancer patients' preferences for written prognostic information provided outside the clinical context. AB - Cancer patients' preferences for written prognostic information independent of the clinical context have not previously been investigated. This study aimed to assist a state cancer organisation to provide information to patients by assessing patients' understanding of statistical information; eliciting their preferences for framing, content and presentation; and assessing the acceptability of a card sort for obtaining preferences. With the exception of conditional and relative survival, initial difficulties in understanding statistical concepts were improved with a plain language explanation. Analysis of the interview transcripts revealed that participants generally supported the provision of written information about survival in booklets and on the Internet. They wanted positive, relevant and clear information. Participants said that the use of, and preferences for, this information would be affected by a patient's age, time since diagnosis, ability to cope with having cancer and the perceived credibility of the information source. They found the card sort acceptable, saying it made the assessment of understanding and selection of preferences easy. This study has identified two fundamental, and sometimes conflicting, factors underlying patients' preferences: the communication of hope and the need to understand information it has also identified patient characteristics thought to influence preferences. These factors and characteristics need to be taken into account when developing written prognostic information for patients. PMID- 14562017 TI - Tyrosinase expression in the peripheral blood of stage III melanoma patients is associated with a poor prognosis: a clinical follow-up study of 110 patients. AB - The aim of this study is to define the relationship between the tyrosinase expression in the peripheral blood and the clinical course of the disease in stage III disease-free melanoma patients after radical lymph node dissection. RT PCR techniques were used to identify tyrosinase mRNA in 110 patients; a total of 542 blood samples were investigated. In all, 54 patients (49%) showed at least one positive result; 13 patients (11.8%) showed baseline positive results: six became negative thereafter, whereas seven showed follow-up positive results until disease progression occurred. One or more positive determinations were found during follow-up in 41 patients with negative baseline tyrosinase. No correlation was found between baseline results and the relapse rate or disease-free survival (DFS), whereas a significant correlation was found between positive tyrosinase results and disease recurrence during follow-up. In fact, 72.9% of positive patients relapsed, but only 19.3% of negative cases did so. The median interval between the positive results and the clinical demonstration of the relapse was 1.9 months (range 1-6.6). Disease-free survival multivariate analysis selected, as independent variables, Breslow thickness (P=0.05), lymph node involvement according to the AJCC classification (P=0.05) and tyrosinase expression (P=0.0001). In conclusion, RT-PCR tyrosinase mRNA expression is a reliable and reproducible marker associated with a high risk of melanoma progression and we encourage its clinical use in routine follow-up. PMID- 14562018 TI - Altered maturation of peripheral blood dendritic cells in patients with breast cancer. AB - Tumours have at least two mechanisms that can alter dendritic cell (DC) maturation and function. The first affects the ability of haematopoietic progenitors to differentiate into functional DCs; the second affects their differentiation from CD14+ monocytes, promoting an early but dysfunctional maturation. The aim of this study was to evaluate the in vivo relevance of these pathways in breast cancer patients. For this purpose, 53 patients with invasive breast cancer were compared to 68 healthy controls. To avoid isolation or culture procedures for enrichment of DCs, analyses were directly performed by flow cytometry on whole-blood samples. The expression of surface antigens and intracellular accumulation of regulatory cytokines upon LPS stimulation were evaluated. The number of DCs, and in particular of the myeloid subpopulation, was markedly reduced in cancer patients (P<0.001). Patient DCs were characterized by a more mature phenotype compared with controls (P=0.016), and had impaired production of IL-12 (P<0.001). These alterations were reverted by surgical resection of the tumour. To investigate the possible role of some tumour-related immunoactive soluble factors, we measured the plasmatic levels of vascular endothelial growth factor, IL-10 and spermine. A significant inverse correlation between spermine concentration and the percentage of DCs expressing IL-12 was found. Evidence was also obtained that in vitro exposure of monocyte-derived DCs to spermine promoted their activation and maturation, and impaired their function. Taken together, our results suggest that both the above-described mechanisms could concomitantly act in breast cancer to affect DC differentiation, and that spermine could be a mediator of dysfunctional maturation of DCs. PMID- 14562019 TI - Hypermethylation of the hTERT promoter inhibits the expression of telomerase activity in normal oral fibroblasts and senescent normal oral keratinocytes. AB - Telomerase activity in human cells closely correlates with the expression of its catalytic subunit, telomerase reverse transcriptase (hTERT). Previously, we reported the lack of telomerase activity in normal human oral fibroblasts (NHOF) and the diminution of telomerase activity during senescence in normal human oral keratinocytes (NHOK). To investigate the underlying mechanisms of telomerase regulation in both cell types, we analysed the expression, promoter activity, and methylation status of the hTERT gene. The expression of hTERT mRNA diminished in senescent NHOK, but was not detected in NHOF at any stage of replication. An exogenous hTERT promoter was active in NHOF and in senescing NHOK, indicating that the lack of hTERT gene expression resulted from alteration of the endogenous hTERT promoter. Since methylation is involved in the silencing of numerous genes, we carried out DNA methylation assays. The assay revealed that the hTERT promoter was hypermethylated in NHOF and was gradually methylated during senescence in NHOK. Treatment of NHOF and senescent NHOK with the demethylating agent 5-aza-2' deoxycytidine restored the expression of endogenous hTERT mRNA. Our results suggest that hypermethylation of the hTERT promoter plays a critical role in the negative regulation of telomerase activity in normal human oral cells. PMID- 14562020 TI - c-Myc alters the DNA damage-induced G2/M arrest in human mammary epithelial cells. AB - Effects of c-Myc overexpression on the DNA damage-induced G2/M checkpoint were studied in finite lifespan, normal human mammary epithelial cells (HMECs). Previously, we showed that c-Myc attenuates G1/S arrest and leads to an inappropriate entry of cells with damaged DNA into the S phase, following treatment with ionising radiation (IR). Here we show that, in striking contrast to control cells, c-Myc-overexpressing HMECs demonstrate a significant attenuation of the G2/M arrest, following IR, and enter into inappropriate mitoses. At the molecular level, ectopic overexpression of c-Myc leads to an unusually high level of expression of cyclin B1, and the elevated levels of cyclin B1 were maintained, after gamma-irradiation. Introduction of DNA damage in c-Myc-overexpressing, normal mammary epithelial cells eventually induces apoptosis, indicating a dramatic sensitisation by c-Myc of DNA damage-induced apoptosis. These two remarkable phenotypes, checkpoint attenuation and sensitisation to apoptosis, resulting from a deregulation of the protooncogene c myc, may produce a unique pattern of alternating cycles, consisting first of amplification of DNA damage, followed by apoptosis-assisted selective pressure. The result of this alternating pattern of damage apoptosis could facilitate the selection of certain genomic alterations required for cellular survival and cellular transformation. PMID- 14562021 TI - Both gene expression for orotate phosphoribosyltransferase and its ratio to dihydropyrimidine dehydrogenase influence outcome following fluoropyrimidine based chemotherapy for metastatic colorectal cancer. AB - Activation of 5-fluorouracil into its nucleotides requires phosphorylation by three pathways involving orotate phosphoribosyl-transferase (OPRT), uridine phosphorylase (UP), or thymidine phosphorylase (TP). In this study, we investigated the association between gene expressions of these three enzymes and antitumour effect. Gene expressions in primary colorectal tumours were analysed by a real-time reverse transcriptional-polymerase chain reaction method in 37 patients receiving oral treatment of tegafur-uracil and leucovorin for metastatic diseases. The median values of OPRT mRNA expressions were 1.39 and 0.85 for responding tumours and nonresponding tumours, respectively, showing a statistically significant difference (P=0.0008). Responding tumours had statistically lower expressions of TP mRNA than nonresponding tumours (P=0.006). However, there was no difference in UP mRNA expression between responding and nonresponding tumours. Patients with high OPRT (>/=1.0) gene expression survived longer than those with low OPRT (<1.0) expression. Dihydropyrimidine dehydrogenase (DPD) gene expressions were measured. Responding tumours had a statistically higher OPRT/DPD ratio than the nonresponding ones (P=0.003). When the median value of the OPRT/DPD ratio was selected as the cutoff value, patients with a high OPRT/DPD ratio survived statistically longer than those with a low ratio (P=0.0014). In conclusion, both the expression of OPRT gene and the OPRT/DPD ratio might be useful as predictive parameters for the efficacy of fluoropyrimidine-based chemotherapy for metastatic colorectal cancer. PMID- 14562023 TI - Glutathione S-transferases (GSTT1 and GSTM1) gene deletions in Tunisians: susceptibility and prognostic implications in breast carcinoma. AB - Glutathione S-transferase Theta1 and Mu1 (GSTT1 and GSTM1) are involved in the metabolism and detoxification of a wide range of potential environmental carcinogens. Conversely, they contribute to tumour cell survival by detoxification of numerous products induced by cancer therapy. The authors designed a large study to investigate the susceptibility and prognostic implications of the GSTT1 and GSTM1 gene deletions in breast carcinoma. The authors used the polymerase chain reaction to characterise the variation of the GSTT1 and GSTM1 genes in 309 unrelated Tunisian patients with breast carcinoma and 242 healthy control subjects. Associations of the clinic-pathologic parameters and the genetic markers with the rates of the breast carcinoma specific overall survival (OVS) and the disease-free survival (DFS) were assessed using univariate and multivariate analyses. A significant association was found between gene deletion of GSTT1 and the risk of early onset of breast carcinoma (OR=1.60, P=0.02). The lack of GSTT1 gene deletion was significantly associated with poor clinical response to chemotherapy (OR=2.29, P=0.03). This association was significantly higher in patients with axillary's lymph node-negative breast carcinoma (OR=12.60, P=0.005). The null-GSTT1 genotype showed a significant association with increased DFS in this selected population of patients. This association was even higher in patients carrying both null-GSTT1 and -GSTM1 genotypes. The gene deletion of GSTs may predict not only the early onset of breast carcinoma but also the clinical response to chemotherapy and the recurrence-free survival for patients with lymph node-negative breast carcinoma. PMID- 14562022 TI - Nuclear shift of hnRNP K protein in neoplasms and other states of enhanced cell proliferation. AB - The heterogeneous nuclear ribonucleoprotein K (hnRNP K), is a ubiquitously expressed protein that interacts with signal transducers, proteins that modulate gene expression and selective RNA and DNA motifs. K protein is modified in response to extracellular signals and directly regulates rates of transcription and translation. We used serum-treated hepatocyte culture, liver after partial hepatectomy and hepatic neoplasms as systems to compare expression, subcellular distribution and tyrosine phosphorylation of K protein in quiescent and dividing cells. The results show that expression of K protein mRNA was increased in states of enhanced proliferation. Levels of nuclear K protein were also higher in proliferating compared to resting cells. In contrast, levels of cytoplasmic K protein were the same or lower in dividing compared to quiescent cells. States of enhanced proliferation were also associated with increased levels of K protein tyrosine phosphorylation. Nuclear shift of K protein in dividing cells may reflect involvement of K protein in signalling multiple events that regulate expression of genes in proliferating cells. PMID- 14562024 TI - Differential SPARC mRNA expression in Barrett's oesophagus. AB - Barrett's oesophagus (BE) is the precursor lesion to adenocarcinoma of the oesophagus. Understanding of the molecular alterations in this multistage process may contribute to improved diagnosis and treatment. Secreted protein acidic and rich in cysteine (SPARC) is a matricellular protein that modulates cell adhesion and growth. Alterations in SPARC expression have been observed in a variety of solid tumours. The aim of this study was to assess the prevalence and timing of SPARC mRNA expression in Barrett's multistage disease and to investigate the impact of SPARC alterations on the development and progression of this disease. SPARC mRNA expression was measured using a quantitative real-time RT-PCR method in 108 specimens from 19 patients with BE without carcinoma, 20 patients with Barrett's-associated adenocarcinoma (EA), and a control group (CG) of 10 patients without evidence of gastro-oesophageal reflux disease. The median SPARC mRNA expression was significantly upregulated in BE tissues compared to paired normal oesophagus (NE) tissues for the BE group (P=0.004) and for the EA group (P<0.001). The SPARC mRNA expression was significantly higher in adenocarcinoma of the oesophagus compared to matching NE tissue and compared to Barrett's tissues in the EA group (P<0.001). Furthermore, SPARC expression values were significantly different between metaplastic and dysplastic Barrett's tissues (P=0.014). In histologically normal squamous oesophagus tissues obtained from carcinoma patients (EA group), the SPARC mRNA expression was significantly higher compared to NE mucosa from the BE group and the CG group (P=0.03). These findings suggest that the upregulation of SPARC mRNA expression is an early event in the development and progression of BE and EA, and that high SPARC expression may be a clinically useful biomarker for the detection of occult adenocarcinoma, and that a widespread 'field effect' is present in the NE of patients with oesophageal adenocarcinoma. PMID- 14562025 TI - ATM variants 7271T>G and IVS10-6T>G among women with unilateral and bilateral breast cancer. AB - Recent reports suggest that two ATM gene mutations, 7271T>G and IVS10-6T>G, are associated with a high risk of breast cancer among multiple-case families. To assess the importance of these two mutations in another 'high-risk' group, young women (under age 51) with multiple primaries, we screened a large population based series of young women with bilateral breast cancer and compared the frequency of these mutations among similar women diagnosed with unilateral breast cancer. The 1149 women included were enrolled in an ongoing population-based case control study of the genetic factors that contribute to bilateral breast cancer; they were not selected on the basis of family history of cancer. Screening for 7271T>G and IVS10-6T>G ATM gene mutations was conducted using DHPLC followed by direct sequencing. The 7271T>G mutation was detected in one out of 638 (0.2%) women with unilateral breast cancer and in none of the bilateral cases, and the IVS10-6T>G mutation in one out of 511 (0.2%) bilateral and in eight out of 638 (1.3%) unilateral breast cancer cases. Carriers of either mutation were not limited to women with a family history. Given the likelihood that young women with bilateral breast cancer have a genetic predisposition, the observed mutation distribution is contrary to that expected if these two mutations were to play an important role in breast carcinogenesis among individuals at high risk. PMID- 14562026 TI - O6-methylguanine-DNA-methyltransferase expression and gene polymorphisms in relation to chemotherapeutic response in metastatic melanoma. AB - In a retrospective study, O(6)-methylguanine-DNA-methyltransferase (MGMT) expression was analysed by immunohistochemistry using monoclonal human anti-MGMT antibody in melanoma metastases in patients receiving dacarbazine (DTIC) as single-drug therapy or as part of combination chemotherapy with DTIC-vindesine or DTIC-vindesine-cisplatin. The correlation of MGMT expression levels with clinical response to chemotherapy was investigated in 79 patients with metastatic melanoma. There was an inverse relationship between MGMT expression and clinical response to DTIC-based chemotherapy (P=0.05). Polymorphisms in the coding region of the MGMT gene were also investigated in tumours from 52 melanoma patients by PCR/SSCP and nucleotide sequence analyses. Single-nucleotide polymorphisms (SNPs) in exon 3 (L53L and L84F) and in exon 5 (I143V/K178R) were identified. There were no differences in the frequencies of these polymorphisms between these melanoma patients and patients with familial melanoma or healthy Swedish individuals. Functional analysis of variants MGMT-I143V and -I143V/K178R was performed by in vitro mutagenesis in Escherichia coli. There was no evidence that these variants decreased the MGMT DNA repair activity compared to the wild-type protein. All melanoma patients with the MGMT 53/84 polymorphism except one had tumours with high MGMT expression. There was no significant correlation between any of the MGMT polymorphisms and clinical response to chemotherapy, although an indication of a lower response rate in patients with SNPs in exon 5 was obtained. Thus, MGMT expression appears to be more related to response to chemotherapy than MGMT polymorphisms in patients with metastatic melanoma. PMID- 14562027 TI - Polymorphisms in the CYP1B1 gene are associated with increased risk of prostate cancer. AB - CYP1B1 has been evaluated as a candidate gene for various cancers because of its function in activating environmental procarcinogens and catalysing the conversion of oestrogens to genotoxic catechol oestrogens. To test the hypothesis that genetic polymorphisms in the CYP1B1 gene may associate with the risk for prostate cancer (CaP), we compared the allele, genotype, and haplotype frequencies of 13 single nucleotide polymorphisms (SNPs) of CYP1B1 among 159 hereditary prostate cancer (HPC) probands, 245 sporadic CaP cases, and 222 unaffected men. When each of the SNPs was analysed separately, marginally significant differences were observed for allele frequencies between sporadic cases and controls for three consecutive SNPs (-1001C/T, -263G/A, and -13C/T, P=0.04-0.07). Similarly, marginally significant differences between sporadic cases and controls in the frequency of variant allele carriers were observed for five consecutive SNPs ( 1001C/T, -263G/A, -13C/T, +142C/G, and +355G/T, P=0.02-0.08). Interestingly, when the combination of these five SNPs was analysed using a haplotype approach, a larger difference was found (P=0.009). One frequent haplotype (C-G-C-C-G of 1001C/T, -263G/A, -13C/T, +142C/G, and +355G/T) was associated with an increased risk for CaP, while the other frequent haplotype (T-A-T-G-T) was associated with a decreased risk for CaP. These findings suggest that genetic polymorphisms in CYP1B1 may modify the risk for CaP. PMID- 14562028 TI - Chromosome aberrations in canine multicentric lymphomas detected with comparative genomic hybridisation and a panel of single locus probes. AB - Recurrent chromosome aberrations are frequently observed in human neoplastic cells and often correlate with other clinical and histopathological parameters of a given tumour type. The clinical presentation, histology and biology of many canine cancers closely parallels those of human malignancies. Since humans and dogs demonstrate extensive genome homology and share the same environment, it is expected that many canine cancers will also be associated with recurrent chromosome aberrations. To investigate this, we have performed molecular cytogenetic analyses on 25 cases of canine multicentric lymphoma. Comparative genomic hybridisation analysis demonstrated between one and 12 separate regions of chromosomal gain or loss within each case, involving 32 of the 38 canine autosomes. Genomic gains were almost twice as common as losses. Gain of dog chromosome (CFA) 13 was the most common aberration observed (12 of 25 cases), followed by gain of CFA 31 (eight cases) and loss of CFA 14 (five cases). Cytogenetic and histopathological data for each case are presented, and cytogenetic similarities with human non-Hodgkin's lymphoma are discussed. We have also assembled a panel of 41 canine chromosome-specific BAC probes that may be used for accurate and efficient chromosome identification in future studies of this nature. PMID- 14562029 TI - The RING-H2 protein RNF11 is overexpressed in breast cancer and is a target of Smurf2 E3 ligase. AB - The breast cancer-associated T2A10 clone was originally isolated from a cDNA library enriched for tumour messenger ribonucleic acids. Our survey of 125 microarrayed primary tumour tissues using affinity purified polyclonal antibodies has revealed that corresponding protein is overexpressed in invasive breast cancer and is weakly expressed in kidney and prostate tumours. Now known as RNF11, the gene encodes a RING-H2 domain and a PY motif, both of which mediate protein-protein interactions. In particular, the PPPPY sequence of RNF11 PY motif is identical to that of Smad7, which has been shown to bind to WW domains of Smurf2, an E3 ubiquitin ligase that mediates the ubiquitination and degradation of the TGFbeta receptor complex. Using various mutants of RNF11 in GST pulldown and immunoprecipitation assays, we found that RNF11 interacts with Smurf2 through the PY motif, leading to ubiquitination of both proteins. Smurf2 plays an active role in the repression of TGFbeta signalling, and our data indicate that overexpression of RNF11, through its interaction with Smurf2, can restore TGFbeta responsiveness in transfected cells. PMID- 14562030 TI - Efficacy of immunoliposomes on cancer models in a cell-surface-antigen-density dependent manner. AB - We have recently established a cancer-reactive human monoclonal antibody, GAH, with a positive ratio of over 90% against stomach cancer. GAH was formulated as polyethyleneglycol (PEG)-modified immunoliposomal doxorubicin (DXR) (ILD) and its efficacy was examined against gastrointestinal human cancers. In in vitro studies, a comparison of ILD with PEG-modified liposomal DXR (LD) demonstrated that ILD had dose-dependent cytotoxicity for GAH-reactive B37 cancer cells, but not LD. In concordance with this result, microscopic observations showed that ILD was bound to and GAH-dependently internalised by B37 cells. In in vivo studies, ILD exhibited significantly greater antitumour activity on cancer xenograft models than LD or free DXR. The relation between efficacy and antigen density was examined on 10 xenograft models bearing cancer cells with varying GAH reactivity. Immunoliposomal doxorubicin therapeutic activity correlated with the antigen density, with a minimum number being required. Also, ILD revealed strong antitumour activity on cancers with low sensitivity to DXR or LD, suggesting that ILD overcame the DXR resistance of antigen-positive cancer cells. Thus, these results show that GAH endows liposomes with targeting activity, resulting in strong efficacy against gastrointestinal cancers. PMID- 14562031 TI - Inhibition of primary colon carcinoma growth and liver metastasis by the A3 adenosine receptor agonist CF101. AB - Adenosine is a purine nucleoside that acts as a regulatory molecule by binding to specific G-protein-coupled A1, A(2A), A(2B), and A3 cell surface receptors. We have recently demonstrated that adenosine inhibits tumour cell growth and concomitantly stimulates bone marrow cell proliferation via activation of the A3 adenosine receptor (A3AR). In the present study, we show that a synthetic agonist to the A3AR, CF101, at the low nanomolar concentration range, inhibits HCT-116 human colon carcinoma cell growth. This effect was reversed by the selective A3AR antagonist MRS1523, demonstrating the specificity of the response. CF101 (given orally) was efficacious in inhibiting the development of primary tumours in xenograft and syngeneic models in which mice were inoculated subcutaneously with human HCT-116 or murine CT-26 colon carcinoma cells, respectively. Moreover, CF101 suppressed (50%, P<0.01) colon cancer liver metastases in syngeneic mice inoculated to the spleen with CT-26 cells. The mechanism of action entailed upregulation of interleukin-12 production in the CF101-treated groups and potentiation of NK cell activity. In the HCT-116 xenograft model in which a combined therapy of CF101 and 5-fluorouracyl (5-FU) was examined, an additive antitumour effect was demonstrated. Moreover, CF101 prevented the 5-FU-induced myelotoxicity, resulting in normal values of white blood cell and neutrophil counts. We conclude that the A3AR agonist CF101, a small orally bioavailable molecule, exerts systemic anticancer, antimetastatic, and myeloprotective effects in colon carcinoma-bearing mice, and may serve as an adjuvant treatment to enhance the chemotherapeutic index and prevent myelotoxicity. PMID- 14562032 TI - Brostallicin (PNU-166196)--a new DNA minor groove binder that retains sensitivity in DNA mismatch repair-deficient tumour cells. AB - Defects in DNA mismatch repair (MMR) are associated with a predisposition to tumorigenesis and with drug resistance owing to high mutation rates and failure to engage DNA-damage-induced apoptosis. DNA minor groove binders (MGBs) are a class of anticancer agents highly effective in a variety of human cancers. Owing to their mode of action, DNA MGB-induced DNA damage may be a substrate for DNA MMR. This study was aimed at investigating the effect of loss of MMR on the sensitivity to brostallicin (PNU-166196), a novel synthetic alpha-bromoacrylic, second-generation DNA MGB currently in Phase II clinical trials and structurally related to distamycin A. Brostallicin activity was compared to a benzoyl mustard derivative of distamycin A (tallimustine). We report that the sensitivities of MLH1-deficient and -proficient HCT116 human colon carcinoma cells were comparable after treatment with brostallicin, while tallimustine resulted in a three times lower cytotoxicity in MLH1-deficient than in -proficient cells. MSH2-deficient HEC59 parental endometrial adenocarcinoma cells were as sensitive as the proficient HEC59+ch2 cells after brostallicin treatment, but were 1.8-fold resistant after tallimustine treatment as compared to the MSH2-proficient HEC59+ch2 counterpart. In addition, p53-deficient mouse fibroblasts lacking PMS2 were as sensitive to brostallicin as PMS2-proficient cells, but were 1.6-fold resistant to tallimustine. Loss of neither ATM nor DNA-PK affected sensitivity to brostallicin in p53-deficient mouse embryonic fibroblasts, indicating that brostallicin-induced cytotoxicity in a p53-deficient genetic background does not seem to require these kinases. These data show that, unlike other DNA MGBs, MMR deficient cells retain their sensitivity to this new alpha-bromoacrylic derivative, indicating that brostallicin-induced cytotoxicity does not depend on functional DNA MMR. Since DNA MMR deficiency is common in numerous types of tumours, brostallicin potentially offers the advantage of being effective against MMR-defective tumours that are refractory to several anticancer agents. PMID- 14562033 TI - Unique induction of p21(WAF1/CIP1)expression by vinorelbine in androgen independent prostate cancer cells. AB - To study the mechanisms of the development of hormone refractory prostate cancer, we established an androgen-independent (AI) prostate cancer cell line derived from hormone-dependent (AD) LNCaP cells. Our previous studies have demonstrated that AI cells are deficient in expression of p21(WAFl/CIP1) (p21) due to overexpressed AR and are resistant to apoptosis. In this study, the induction of p53 and p21 expression by vinorelbine (Navelbine) was compared between AD and AI cells in an attempt to understand the difference(s) in apoptotic signalling pathways in these cells. Using a series of deletion of p21 reporter constructs, we found that vinorelbine mediated p21 induction in a p53-dependent manner in AD cells. In contrast, p21 expression restored by vinorelbine in AI cells was found to be through both p53-dependent and-independent pathways. In the absence of two p53 binding sites, Spl-3 and Spl-4 sites, in the promoter of human p21 gene, were found to be required for vinorelbine-mediated p21 activation. No p21 induction was observed by paclitaxel in AI cells. Exposure of AI cells to paciltaxel followed by vinorelbine produced synergism. Our data, thus, provide a basis for the synergistic combination of vinorelbine and paclitaxel for the treatment of advanced prostate cancer. PMID- 14562034 TI - Cathepsin-B and cathepsin-L expression levels do not correlate with sensitivity of tumour cells to TNF-alpha-mediated apoptosis. AB - Recently, evidence has been accumulated that besides the caspase proteases, lysosomal cathepsins may play a role in apoptosis induction. This is especially significant as many human tumour cells express high levels of cathepsins, which might sensitise these cells to specific proapoptotic stimuli mediated by cathepsins. We found that TNF-alpha-mediated DNA fragmentation in tumour cells was significantly reduced in the presence of E64d and CA074Me, two inhibitors of lysosomal cysteine proteases. Transient transfection of cathepsin-B (Cath-B) and L (Cath-L) resulting in expression levels comparable to those found in many tumours did not sensitise tumour cells to TNF-alpha-mediated apoptosis. As lysosomal proteases are thought to be activated by their release from this organelle into the cytosol, we used the lysosomotropic detergent N-dodecyl imidazole-HCl (NDI-HCl) to disturb lysosomal integrity efficiently and trigger the release of its proteolytic content into the cytosol. Treatment of HeLa cells with NDI-HCl resulted in cell death, which, however, could also not be influenced by augmented Cath-B or -L expression levels. Therefore, our data do not support the hypothesis that the high Cath-B or -L expression levels frequently detected in tumour cells might be exploited to target selectively those tumours for an enhanced cell death effect induced by lysosomotropic agents. PMID- 14562035 TI - The expression of P-glycoprotein does influence the distribution of novel fluorescent compounds in solid tumour models. AB - Solid tumours display a complex drug resistance phenotype that involves inherent and acquired mechanisms. Multicellular resistance is an inherent feature of solid tumours and is known to present significant barriers to drug permeation in tumours. Given this barrier, do acquired resistance mechanisms such as P glycoprotein (P-gp) contribute significantly to resistance? To address this question, the multicellular tumour spheroid (MCTS) model was used to examine the influence of P-gp on drug distribution in solid tissue. Tumour spheroids (TS) were generated from either drug-sensitive MCF7(WT) cells or a drug-resistant, P gp-expressing derivative MCF7(Adr). Confocal microscopy was used to measure time courses and distribution patterns of three fluorescent compounds; calcein-AM, rhodamine123 and BODIPY-taxol. These compounds were chosen because they are all substrates for P-gp-mediated transport, exhibit high fluorescence and are chemically dissimilar. For example, BODIPY-taxol and rhodamine 123 showed high accumulation and distributed extensively throughout the TS(WT), whereas calcein AM accumulation was restricted to the outermost layers. The presence of P-gp in TS(Adr) resulted in negligible accumulation, regardless of the compound. Moreover, the inhibition of P-gp by nicardipine restored intracellular accumulation and distribution patterns to levels observed in TS(WT). The results demonstrate the effectiveness of P-gp in modulating drug distribution in solid tumour models. However, the penetration of agents throughout the tissue is strongly determined by the physico-chemical properties of the individual compounds. PMID- 14562036 TI - Bax is essential for mitochondrion-mediated apoptosis but not for cell death caused by photodynamic therapy. AB - The role of Bax in the release of cytochrome c from mitochondria and the induction of apoptosis has been demonstrated in many systems. Using immunocytochemical staining, we observed that photodynamic therapy (PDT) with the photosensitiser Pc 4 induced Bax translocation from the cytosol to mitochondria, and the release of cytochrome c from mitochondria as early signalling for the intrinsic pathway of apoptosis in human breast cancer MCF-7c3 cells. To test the role of Bax in apoptosis, MCF-7c3 cells were treated with Bax antisense oligonucleotides, which resulted in as much as a 50% inhibition of PDT-induced apoptosis. In the second approach, Bax-negative human prostate cancer DU-145 cells were studied. Following PDT, the hallmarks of apoptosis, including the release of cytochrome c from mitochondria, loss of mitochondrial membrane potential, caspase activation, and chromatin condensation and fragmentation, were completely blocked in these cells. Restoration of Bax expression in DU-145 cells restored apoptosis, indicating that the resistance of DU-145 cells to PDT-induced apoptosis is due to the lack of Bax rather than to another defect in the apoptotic machinery. However, despite the inhibition of apoptosis, the Bax negative DU-145 cells were as photosensitive as Bax-replete MCF-7c3 cells, as determined by clonogenic assay. Thus, for Pc 4-PDT, the commitment to cell death occurs prior to Bax activation. PMID- 14562038 TI - The Chfr mitotic checkpoint protein functions with Ubc13-Mms2 to form Lys63 linked polyubiquitin chains. AB - We recently described a novel checkpoint pathway that functions early in mitosis to delay chromosome condensation in response to microtubule poisons. The only gene implicated so far in this checkpoint pathway is chfr, whose protein product contains a RING domain and has ubiquitin ligase activity in vitro. The significance of this activity in vivo is unclear. A recent report suggested that the Chfr protein targets itself for proteasome-dependent degradation in mitotic cells through autoubiquitination. However, we observe that in mitosis Chfr exhibits a phosphorylation-dependent electrophoretic mobility shift with no change in overall protein levels. Further analysis of its ubiquitin ligase activity revealed that Chfr can catalyse the formation of noncanonical Lys63 linked polyubiquitin chains with Ubc13-Mms2 acting as the ubiquitin-conjugating enzyme. Ubc13-Mms2 and Lys63-polyubiquitin chains are not associated with targeting proteins to the proteasome, but rather with signaling cellular stress. We propose that Chfr may have a role in signaling the presence of mitotic stress induced by microtubule poisons. PMID- 14562039 TI - Proteasome inhibitors potentiate leukemic cell apoptosis induced by the cyclin dependent kinase inhibitor flavopiridol through a SAPK/JNK- and NF-kappaB dependent process. AB - Interactions between proteasome and cyclin-dependent kinase inhibitors have been examined in human leukemia cells in relation to induction of apoptosis. Simultaneous exposure (24 h) of U937 myelomonocytic leukemia cells to 100 nM flavopiridol and 300 nM MG-132 resulted in a marked increase in mitochondrial injury (cytochrome c, Smac/DIABLO release, loss of deltaPsi(m)), caspase activation, and synergistic induction of cell death, accompanied by a marked decrease in clonogenic potential. Similar effects were observed with other proteasome inhibitors (e.g., Bortezomib (VELCADE trade mark bortezomib or injection), lactacystin, LLnL) and cyclin-dependent kinase inhibitors (e.g., roscovitine), as well as other leukemia cell types (e.g., HL-60, Jurkat, Raji). In U937 cells, synergistic interactions between MG-132 and flavopiridol were associated with multiple perturbations in expression/activation of signaling- and survival-related proteins, including downregulation of XIAP and Mcl-1, activation of JNK and p34(cdc2), and diminished expression of p21(CIP1). The lethal effects of MG-132/flavopiridol were not reduced in leukemic cells ectopically expressing Bcl-2, but were partially attenuated in cells ectopically expressing dominant negative caspase-8 or CrmA. Flavopiridol/proteasome inhibitor-mediated lethality was also significantly diminished by agents and siRNA blocking JNK activation. Lastly, coadministration of MG-132 with flavopiridol resulted in diminished DNA binding of NF-kappaB. Notably, pharmacologic interruption of the NF-kappaB pathway (e.g., by BAY 11-7082, PDTC, or SN-50) or molecular dysregulation of NF kappaB (i.e., in cells ectopically expressing an IkappaBalpha super-repressor) mimicked the actions of proteasome inhibitors in promoting flavopiridol-induced mitochondrial injury, JNK activation, and apoptosis. Together, these findings indicate that proteasome inhibitors strikingly lower the apoptotic threshold of leukemic cells exposed to pharmacologic CDK inhibitors, and suggest that interruption of the NF-kappaB cytoprotective pathway and JNK activation both play key roles in this phenomenon. They also raise the possibility that combining proteasome and CDK inhibitors could represent a novel antileukemic strategy. PMID- 14562040 TI - Association of a functional tandem repeats in the downstream of human telomerase gene and lung cancer. AB - Chemoprevention has been widely explored as a promising strategy for controlling the incidence of lung cancer, the leading cause of cancer-related death. To maximize the benefit of lung cancer chemoprevention, it is important to identify individuals at high risk for the disease. The genetic background has been shown to play an important role in one's risk of developing lung cancer. We report here the identification of a polymorphic tandem repeats minisatellite (termed MNS16A) in the downstream region of the human telomerase gene. This minisatellite is located upstream of an antisense transcript from the human telomerase gene locus and was demonstrated to have promoter activity. The promoter activity was significantly lower in the construct containing the shorter repeats, suggesting that the MNS16A variant may have a relevance of functionality. To explore the role of this novel polymorphism in lung cancer, we conducted a pilot hospital based case-control study by identifying the MNS16A genotype with genomic DNA from 53 lung cancer patients and 72 cancer-free controls. We found four different alleles and classified them as shorter (S) or longer (L) on the functional basis of the length of the repeats in the controls. The MNS16A genotype distributions of the SS, SL, and LL genotypes were 11, 32, and 57%, respectively, in the cases, and 14, 40, and 46%, respectively, in the controls. Compared with the SS+SL genotype, the LL genotype was associated with greater than twofold increased risk of lung cancer (odds ratio=2.18; 95% confidence interval=0.92, 5.20) after adjustment for age, sex, ethnicity, and smoking status, suggesting a potential role of MNS16A in lung cancer susceptibility. Larger studies are needed to verify our findings. PMID- 14562041 TI - MBD4 deficiency reduces the apoptotic response to DNA-damaging agents in the murine small intestine. AB - MBD4 was originally identified through its methyl binding domain, but has more recently been characterized as a thymine DNA glycosylase that interacts with the mismatch repair (MMR) protein MLH1. In vivo, MBD4 functions to reduce the mutability of methyl-CpG sites in the genome and mice deticient in MBD4 show increased intestinal tumorigenesis on an Apc(Min/+) background. As MLH1 and other MMR proteins have been functionally linked to apoptosis, we asked whether MBD4 also plays a role in mediating the apoptotic response within the murine small intestine. Mice deficient for MBD4 showed significantly reduced apoptotic responses 6 h following treatment with a range of cytotoxic agents including gamma-irradiation, cisplatin, temozolomide and 5-fluorouracil (5-FU). This leads to increased clonogenic survival in vivo in Mbd4(-/-) mice following exposure to either 5-FU or cisplatin. We next analysed the apoptotic response to 5-FU and temozolomide in doubly mutant Mbd4(-/-), Mlh1(-/-) mice but observed no additive decrease. The results imply that MBD4 and MLH1 lie in the same pathway and therefore that MMR-dependent apoptosis is mediated through MBD4. MBD4 deficiency also reduced the normal apoptotic response to gamma-irradiation, which we show is independent of Mlh1 status (at least in the murine small intestine), so suggesting that the reliance upon MBD4 may extend beyond MMR-mediated apoptosis. Our results establish a novel functional role for MBD4 in the cellular response to DNA damage and may have implications for its role in suppressing neoplasia. PMID- 14562042 TI - Ras oncogene directs expression of a differentially sialylated, functionally altered beta1 integrin. AB - Intense investigation has centered on understanding the regulation of integrin cell adhesion receptors. In the present study, we propose that variant N glycosylation represents an important mechanism for regulation of beta1, but not beta3 or beta5 integrins. We find that expression of oncogenic ras in HD3 colonocytes causes increased alpha2-6 sialylation of beta1 integrins, whereas expression of dominant-negative ras induces decreased alpha2-6 sialylation, relative to cells with wild-type ras. In contrast, neither beta3 nor beta5 integrins are alpha2-6 sialylated, regardless of the state of ras activation. Results from RT-PCR analyses suggest that differential integrin sialylation is due to a ras-dependent alteration in the expression of ST6Gal I, the enzyme that adds alpha2-6-linked sialic acids. Cells that express differentially sialylated beta1 integrins exhibit altered adhesion to collagen I (a beta1 ligand), but not to vitronectin (a beta3 or beta5 ligand). Similarly, the enzymatic removal of cell surface sialic acids from control cells alters binding to collagen, but not to vitronectin. Finally, using a cell-free receptor/ligand-binding assay, we show that purified, desialylated alpha1beta1 integrins have diminished collagen binding capability, providing strong evidence that sialic acids play a causal role in regulating beta1 integrin function. PMID- 14562043 TI - Role of the RNA-binding protein HuR in colon carcinogenesis. AB - Immunohistochemical analysis of paired tumor and normal tissue specimens revealed that the expression and cytoplasmic abundance of the RNA-binding protein HuR increased with malignancy, particularly in colon carcinomas. Interventions to modulate HuR expression in human RKO colon cancer cells altered gene expression profiles and identified beta-catenin mRNA as a novel HuR target. Subcutaneous injection of HuR-overexpressing RKO cells into nude mice produced significantly larger tumors than those arising from control populations; conversely, RKO cells expressing reduced HuR through small interference RNA- or antisense HuR-based approaches developed significantly more slowly. We propose that HuR-regulated target mRNA expression contributes to colon cancer growth. Our results suggest a pivotal function for HuR in colon carcinogenesis. PMID- 14562044 TI - Expression profiling of epithelial plasticity in tumor progression. AB - Epithelial-to-mesenchymal transition (EMT), a switch of polarized epithelial cells to a migratory, fibroblastoid phenotype, is increasingly considered as an important event during malignant tumor progression and metastasis. To identify molecular players involved in EMT and metastasis, we performed expression profiling of a set of combined in vitro/in vivo cellular models, based on clonal, fully polarized mammary epithelial cells. Seven closely related cell pairs were used, which were modified by defined oncogenes and/or external factors and showed specific aspects of epithelial plasticity relevant to cell migration, local invasion and metastasis. Since mRNA levels do not necessarily reflect protein levels in cells, we used an improved expression profiling method based on polysome-bound RNA, suitable to analyse global gene expression on Affymetrix chips. A substantial fraction of all regulated genes was found to be exclusively controlled at the translational level. Furthermore, profiling of the above multiple cell pairs allowed one to identify small numbers of genes by cluster analysis, specifically correlating gene expression with EMT, metastasis, scattering and/or oncogene function. A small set of genes specifically regulated during EMT was identified, including key regulators and signaling pathways involved in cell proliferation, epithelial polarity, survival and trans differentiation to mesenchymal-like cells with invasive behavior. PMID- 14562045 TI - An increase in the expression and total activity of endogenous p60(c-Src) in several factor-independent mutants of a human GM-CSF-dependent leukemia cell line (TF-1). AB - Growth factor independence of hematopoietic cells can be induced by ectopic expression of a variety of oncogenes encoding receptor or cytoplasmic tyrosine kinases. To examine whether the activation of tyrosine kinases occurs in factor independent mutants in vivo, the tyrosine-phosphorylated proteins from 14 factor independent mutants of a GM-CSF-dependent cell line (TF-1) were analysed. These mutants did not secrete any growth-stimulating activity for TF-1 cells, suggesting that activation of intracellular signaling rather than an autocrine stimulation by secreted growth factors is responsible for their factor independent growth. In 11 out of 14 GM-CSF-independent mutants analysed, a constitutively tyrosine-phosphorylated protein of 60 kDa was detected, which was subsequently identified as p60(c-Src). The kinase activity of p60(c-Src) was increased up to 12-fold in these mutants, which was at least in part due to overexpression of the c-src gene on the RNA and protein level. The Src substrate Sam68 showed an increased phosphorylation in mutants with high Src activity, suggesting that p60(c-Src) triggers downstream signaling in these cells. Treatment of the factor-independent mutants with the Src kinase inhibitor PP2 resulted in a reduced proliferation, demonstrating that Src kinases are essential for these cells for maximal proliferation. Further analysis of factor-independent mutants with low or undetectable Src activity revealed a constitutive phosphorylation of the common beta chain of the GM-CSF receptor and STAT5. Our data indicate an increase in the expression and total activity of endogenous p60(c-Src) in several GM-CSF-independent TF-1 mutants, further underlining the role of Src in the process of autonomous growth of hematopoietic cells. PMID- 14562046 TI - Epstein-Barr virus can inhibit genotoxin-induced G1 arrest downstream of p53 by preventing the inactivation of CDK2. AB - Epstein-Barr virus (EBV) is involved in the pathogenesis of several B cell lymphoproliferations, but the precise contribution it makes to the aetiology of each remains unclear. In vitro, the virus has potent growth transforming activity and efficiently induces the continuous proliferation of normal human B cells. A comparison of EBV-infected primary B cells with an isogenic population induced to proliferate by CD40-ligand (CD40L) and IL4 has revealed that EBV can override - by a novel mechanism - the p53/pRb-mediated G1 checkpoint activated in normal B cells by a genotoxic stress. In cells responding to cisplatin, although p53 is stabilized and activated, EBV latent gene expression appears to inhibit the accumulation of newly synthesized p21(WAF1/CIP1) and the downregulation of cyclin D2 that occur in the normal cells. Consequently, in the EBV-infected cells, CDK2 remains active, hyperphosphorylation of pRb is maintained and the replication of damaged DNA can occur. Under conditions of severe genomic stress, this absence of p21(WAF1/CIP1) function can result in apoptosis; however, when damage is less sustained, genomic instability may arise and this in turn could contribute to the development of a variety of EBV-associated B cell malignancies. PMID- 14562047 TI - Functional and molecular characterization of a KIR3DL2/p140 expressing tumor specific cytotoxic T lymphocyte clone infiltrating a human lung carcinoma. AB - T lymphocytes infiltrating a human lung carcinoma stimulated in vitro with autologous tumor cell line showed a TCRVbeta13.6(+) T-cell expansion. This subset was isolated using TCRVbeta-specific antibody and several T-cell clones were generated. All these clones expressed a unique Vbeta13.6-Jbeta2.7 TCR with the same junctional region strongly suggesting that they derived from the same cell. They were CD8(+)/CD28(-) and expressed the MHC class I binding killer cell Ig like receptor (KIR)3DL2/p140, but not KIR3DL1/p70, KIR2DL1/p58.1 and KIR2DL2/3/p58.2. Sequence analysis indicated that KIR3DL2/p140 cDNA was identical to the previously reported 3DL2*002 allele except for two nucleic acid substitutions. Functional studies showed that KIR3DL2/p140(+) CTL secrete a significant level of IFNgamma and mediate an HLA-A2-restricted cytotoxicity against the autologous and some allogeneic tumor cells but not towards the autologous EBV-B cells. Strikingly, both the lytic and the cytokine secretion activities induced upon specific cell interactions were unaffected by anti KIR3DL2/p140 antibody. In addition, crosslinking KIR3DL2/p140 molecules on CTL did not result into the modification of cytotoxicity and cytokine production triggered by anti-CD3 antibody. These results strongly suggest that, as opposed to distinct KIR expressed by CTL, the in vitro KIR3DL2/p140 engagement does not result into inhibitory (nor activatory) effects on tumor-specific CTL. PMID- 14562048 TI - Functional and molecular characterization of the epithelioid to round transition in human colorectal cancer LoVo cells. AB - In subclones of the human colon cancer LoVo cell line, there is a reproducible spontaneous transition from an epithelioid (E) to a round (R) morphotype. The E to R transition is associated with increased cell growth, absence of E-cadherin dependent compaction in a slow aggregation assay, loss of contact inhibition of motility and directional migration in a wound filling motility assay. Furthermore, none of the E subclones from LoVo was invasive into chick heart fragments. This is in contrast to the R subclones that were either nonadherent or adherent and invasive. Macroarray analysis demonstrated transcriptional downregulation of plakoglobin in R type LoVo cells and this was confirmed at the level of the mRNA by quantitative RT-PCR. Western blotting showed lower expression of all components of the E-cadherin/catenin complex in R subclones. Interestingly, treatment of R subclones with the demethylating agent 5-aza-2' deoxycytidine resulted in restoration of the E morphotype, higher expression of E cadherin, but not plakoglobin mRNA, and higher expression of E-cadherin and plakoglobin at the protein level. PMID- 14562049 TI - Mechanisms of transcriptional regulation by Rb-E2F segregate by biological pathway. AB - The E2F family of transcription factors are critical regulators of the cell cycle and have also been implicated in apoptosis, development, DNA damage checkpoints, and differentiation. Retinoblastoma (Rb) proteins interact with E2F to regulate transcription, and several mechanisms have been proposed for Rb-E2F transcriptional regulation. We designed microarray-based experiments to characterize the relative contributions of each mechanism, and unexpectedly, we found that distinct functional gene groups show preference for one mechanism over the others. We propose that such a distribution may provide signaling specificity to enable regulatory proteins to turn on or off entire pathways that determine cell fate. PMID- 14562050 TI - Activation of the Wnt pathway in non small cell lung cancer: evidence of dishevelled overexpression. AB - Non small cell lung cancer (NSCLC) is the leading cause of cancer deaths in the United States and worldwide. Unfortunately, standard therapies remain inadequate. An increased understanding of the molecular biology of lung cancer biology is required to develop more effective new therapies. In this report, we show that the Wnt pathway is activated through Dishevelled (Dvl) overexpression in NSCLC. Analysis of freshly resected tumors and lung cancer cell lines demonstrate that Dvl-3, a critical mediator of Wnt signaling, is overexpressed. Specifically, Dvl 3 was overexpressed significantly in 75% of fresh NSCLC microdissected samples compared to control paired matched normal lung samples. To evaluate the biological significance of Wnt signaling and, in particular, Dvl function in lung cancer, we transfected siRNA (designed to inhibit selectively human Dvl-1, -2, and -3), to the NSCLC cell line H1703, which is known to have beta-catenin mediated Tcf-dependent transcriptional activity. Here, we demonstrate that Dvl specific siRNA treatment in H1703 decreases significantly Dvl and beta-catenin expression, resulting in reduction of Tcf-dependent transcriptional activity, and, importantly, growth inhibition. Taken together, these data support the novel hypothesis that Dvl overexpression is critical to Wnt signaling activation and cell growth in NSCLC. PMID- 14562051 TI - Tissue concentrations of platelet-activating factor in colorectal carcinoma: inverse relationships with Dukes' stage of patients. AB - The lipid mediator platelet-activating factor (PAF) plays a role in cancer. We investigated its presence in human colon carcinoma by assessing the levels of tissue phospholipase A(2) (PLA(2), the key enzyme in the generation of the lyso PAF precursor), lyso-PAF, PAF and acetylhydrolase activity (AHA, the key enzyme in PAF degradation) in colorectal cancer patients and by correlating them with Dukes' classification. The results highlighted that the tumour tissues of Dukes' A and B patients had significantly higher PLA(2), lyso-PAF, PAF and AHA levels as compared with nontumour tissues. Dukes' C patients had higher PLA(2), lyso-PAF and AHA levels but unchanged PAF. Dukes' D patients had higher AHA levels but unchanged PLA(2), lyso-PAF and PAF. A pathophysiological role for PAF is suggested in human colon carcinoma. PMID- 14562052 TI - Characterization of novel human ovarian cancer-specific transcripts (HOSTs) identified by serial analysis of gene expression. AB - A better understanding of changes in gene expression during ovarian tumorigenesis and the identification of specific tumor markers may lead to novel strategies for diagnosis and therapy for this disease. Using our serial analysis of gene expression (SAGE) data, as well as public SAGE databases that contained a total of 137 SAGE libraries representing a wide variety of normal and neoplastic tissues, we identified five novel SAGE tags specifically expressed in ovarian cancer. Database analysis, cloning and, sequencing of the corresponding expressed sequence tags revealed details about these transcripts that we named human ovarian cancer-specific transcripts (HOSTs). HOST1 was found to be identical to the gene encoding ovarian marker CA125 (MUC16). HOST2 is a novel gene containing multiple copies of retroviral-related sequences without an obvious open reading frame. HOST3 encodes the tight-junction protein claudin-16 (CLDN16). HOST4 encodes a poorly characterized proteoglycan link protein (LP), and HOST5 codes for a type II sodium-dependent phosphate transporter (SLC34A2). Except for MUC16, these genes have not previously been shown to be expressed in ovarian or other cancers. Northern blot analysis confirmed that HOST genes are rarely expressed in normal tissues or nonovarian cancers, but are frequently expressed in ovarian cancer-derived cell lines and primary tumors. Moreover, HOST genes are upregulated in all four major subtypes of ovarian cancer compared to cultivated ovarian surface epithelial cells, as concluded by real-time reverse transcription (RT)-PCR using a panel of microdissected ovarian tumors. The sodium-dependent phosphate transporter (HOST5/SLC34A2) expression was associated with increased differentiation in ovarian serous tumors. While the roles of HOSTs in ovarian malignant transformation remain unclear, we propose that HOSTs may represent alternative targets for diagnosis and therapy and of this deadly disease. PMID- 14562053 TI - Preferential integration of human papillomavirus type 18 near the c-myc locus in cervical carcinoma. AB - The development of cervical cancer is highly associated with human papillomavirus (HPV) infection. Greater than 99% of all cervical tumors contain HPV DNA. Integration of high-risk HPV has been temporally associated with the acquisition of a malignant phenotype. Recent work from our lab has shown that HPV16, the most common high-risk HPV associated with cervical carcinoma, preferentially integrates at loci containing human common fragile sites (CFSs). CFSs are regions of genomic instability that have also been associated with deletions, translocations, and gene amplification during cancer development. The current work shows that HPV18, the second most prevalent high-risk HPV type found in cervical tumors, preferentially targets the CFSs. We identified 27 unique HPV18 integrations in cervical tumors, of which 63% (P<0.001) occur in CFSs. However, the distribution of HPV18 integrations found were profoundly different from those found for HPV16. Specifically, 30% of all HPV18 integrations occurred within the chromosomal band 8q24 near the c-myc proto-oncogene. None of the HPV16 integrations occurred in this region. Previous low-resolution mapping suggested that c-myc may be a target of HPV integration. Our data at nucleotide resolution confirm that in HPV18-positive cervical tumors, the region surrounding c-myc is indeed a hot spot of viral integration. These results demonstrate that CFSs are preferred sites of integration for HPV18 in cervical tumors. In addition, we have identified multiple cellular genes that have been disrupted by HPV18 integration in cervical tumors. Our results suggest that the sites of HPV18 integration are nonrandom and may play an important role in the development of cervical tumors. PMID- 14562054 TI - Increased expression and no mutation of the Flap endonuclease (FEN1) gene in human lung cancer. AB - The underlying molecular mechanisms leading to microsatellite alteration and mutations in human lung cancer remain unknown. Since Flap endonuclease1 (Fen1), which functions in the base excision repair system, has been shown to be involved in tumor progression of mouse models with microsatellite instability in a haplo insufficient manner, we performed expression and mutation analyses for FEN1 in human lung cancer cell lines. Reverse transcriptase PCR analysis revealed that all 49 lung cancer cell lines (20 small cell lung cancers (SCLCs) and 29 non small cell lung cancers (NSCLCs)) expressed FEN1. In addition, microarray analysis showed that FEN1 expression was elevated significantly by 1.65-fold (P=0.001) in SCLC cell lines compared to normal lung controls (normal human lung cultures and immortalized normal human bronchial epithelial cell lines). FEN1 protein was abundantly expressed in all 23 lung cancer cell lines (10 SCLCs and 13 NSCLCs) and was expressed at lower levels in three of four normal lung epithelial culture controls. Direct sequencing of genomic DNAs revealed no FEN1 mutation in seven SCLCs and nine NSCLCs. As part of this analysis we discovered and sequenced a FEN1 pseudogene (GenBank accession #AY249897) located at 1p22.2. This pseudogene is amplified from cDNA preparations contaminated with genomic DNA and must be taken into account in any FEN1 mutation analysis studies. Our results suggest that alterations of FEN1 are not likely to contribute to development of lung cancer. PMID- 14562055 TI - Cloning and characterization of a novel gene PDRG that is differentially regulated by p53 and ultraviolet radiation. AB - We report the cloning and characterization of a novel p53 and DNA damage regulated gene (PDRG). The human and mouse PDRG sequences are highly homologous and contain open reading frames of 133 amino acids each with molecular masses of 15.5 and 15.3 kDa, respectively. PDRG codes for a novel protein that does not show similarity to any known protein in the databases. Human PDRG is predominantly expressed in normal testis and exhibits reduced but detectable expression in other organs. GFP-tagged PDRG was predominantly detected as aggregates that appeared to reside in a distinct subcellular compartment. PDRG mRNA was upregulated by ultraviolet radiation (UV) but downregulated by tumor suppressor p53. UV is known to transcriptionally upregulate the expression of certain genes by activating the transcription factor Oct-1, while p53 has been reported to suppress transcription of certain genes by directly binding to a novel head-to-tail response element. Cloning and sequence analysis of PDRG promoter revealed the presence of Oct-1-binding element and a putative head-to tail-type p53-binding site. Indeed, UV as well as exogenous Oct-1 independently increased PDRG promoter activity, suggesting that UV could mediate PDRG upregulation via Oct-1. Exogenous wild-type p53 was found to downregulate the PDRG promoter activity indicating that wild-type p53 transcriptionally suppresses the expression of PDRG and may mediate its effect via the putative head-to-tail response element. Furthermore, stable expression of exogenous PDRG was found to decrease the clonogenic survival after UV irradiation, which highlights the significance of PDRG in facilitating UV-induced killing. PMID- 14562056 TI - Five new mouse susceptibility to colon cancer loci, Scc11-Scc15. AB - Although several genes causing familial cancer syndromes have been identified, susceptibility to sporadic cancer remains unsolved. Animal experiments have demonstrated a large number of quantitative trait loci affecting cancer susceptibility. Previously, we described in mouse strain CcS-19/Dem five susceptibility to colon cancer (Scc) loci, Scc1-Scc5 controlling tumor numbers. In the present study, we performed an independent identical mouse cross using a distinct carcinogen, azoxymethane, to induce colon tumors. We confirmed all five originally described Scc loci and detected five additional new Scc loci; Scc11 Scc15. All these loci were detected in two-way interactions. PMID- 14562057 TI - Stable reprogrammed heterokaryons form spontaneously in Purkinje neurons after bone marrow transplant. AB - Heterokaryons are the product of cell fusion without subsequent nuclear or chromosome loss. Decades of research using Sendai-virus or polyethylene glycol (PEG)-mediated fusion in tissue culture showed that the terminally differentiated state of a cell could be altered. But whether stable non-dividing heterokaryons could occur in animals has remained unclear. Here, we show that green fluorescent protein (GFP)-positive bone-marrow-derived cells (BMDCs) contribute to adult mouse Purkinje neurons through cell fusion. The formation of heterokaryons increases in a linear manner over 1.5 years and seems to be stable. The dominant Purkinje neurons caused the BMDC nuclei within the resulting heterokaryons to enlarge, exhibit dispersed chromatin and activate a Purkinje neuron-specific transgene, L7-GFP. The observed reprogrammed heterokaryons that form in brain may provide insights into gene regulation associated with cell-fate plasticity. PMID- 14562059 TI - Positioning of follicular dendritic cells within the spleen controls prion neuroinvasion. AB - Peripheral infection is the natural route of transmission in most prion diseases. Peripheral prion infection is followed by rapid prion replication in lymphoid organs, neuroinvasion and progressive neurological disease. Both immune cells and nerves are involved in pathogenesis, but the mechanisms of prion transfer from the immune to the nervous system are unknown. Here we show that ablation of the chemokine receptor CXCR5 juxtaposes follicular dendritic cells (FDCs) to major splenic nerves, and accelerates the transfer of intraperitoneally administered prions into the spinal cord. Neuroinvasion velocity correlated exclusively with the relative locations of FDCs and nerves: transfer of CXCR5-/- bone marrow to wild-type mice induced perineural FDCs and enhanced neuroinvasion, whereas reciprocal transfer to CXCR5-/- mice abolished them and restored normal efficiency of neuroinvasion. Suppression of lymphotoxin signalling depleted FDCs, abolished splenic infectivity, and suppressed acceleration of pathogenesis in CXCR5-/- mice. This suggests that prion neuroimmune transition occurs between FDCs and sympathetic nerves, and relative positioning of FDCs and nerves controls the efficiency of peripheral prion infection. PMID- 14562058 TI - Discs-Large and Strabismus are functionally linked to plasma membrane formation. AB - During early embryogenesis in Drosophila melanogaster, extensive vesicle transport occurs to build cell boundaries for 6,000 nuclei. Here we show that this important process depends on a functional complex formed between the tumour suppressor and adaptor protein Discs-Large (Dlg) and the integral membrane protein Strabismus (Stbm)/Van Gogh (Vang). In support of this idea, embryos with mutations in either dlg or stbm displayed severe defects in plasma membrane formation. Conversely, overexpression of Dlg and Stbm synergistically induced excessive plasma membrane formation. In addition, ectopic co-expression of Stbm (which associated with post-Golgi vesicles) and the mammalian Dlg homologue SAP97/hDlg promoted translocation of SAP97 from the cytoplasm to both post-Golgi vesicles and the plasma membrane. This effect was dependent on the interaction between Stbm and SAP97. These findings suggest that the Dlg-Stbm complex recruits membrane-associated proteins and lipids from internal membranes to sites of new plasma membrane formation. PMID- 14562060 TI - Coping with peer rejection. PMID- 14562061 TI - Health chiefs poised to step up US scrutiny of microbe research. PMID- 14562062 TI - Routine tests reveal unknown strains of BSE prions. PMID- 14562063 TI - Physician launches public protest over medical Nobel. PMID- 14562064 TI - Earthquake makes waves for tsunami models. PMID- 14562065 TI - Bleak forecast for space weather. PMID- 14562066 TI - Letters reveal scale of German agency's Nazi corruption. PMID- 14562067 TI - Congress split over funding for 'safe' nuclear reactor. PMID- 14562068 TI - Channel hoppers land chemistry Nobel. PMID- 14562069 TI - Economics prize for duo who resolved random results. PMID- 14562071 TI - Time to choose. PMID- 14562072 TI - Damned if they do, damned if they don't... PMID- 14562073 TI - JET at risk if Europe can not afford to pay for ITER. PMID- 14562074 TI - Pointless suffering of animals can be avoided. PMID- 14562075 TI - Effective protection may alter the look of Venice. PMID- 14562076 TI - How do impact factors relate to the real world? PMID- 14562080 TI - Auxetic materials: avoiding the shrink. PMID- 14562081 TI - Biogeography: the coelacanth of frogs. PMID- 14562082 TI - Earthquakes: good tidings. PMID- 14562083 TI - Proteomics: where's Waldo in yeast? PMID- 14562084 TI - Solar physics: heat exposure. PMID- 14562085 TI - Prion diseases: a nucleic-acid accomplice? PMID- 14562086 TI - Materials science: zero-expansion plan. PMID- 14562088 TI - Evolution: opportunity versus innovation. PMID- 14562089 TI - Astronomy: faking it. PMID- 14562091 TI - Evolution: single-gene speciation by left-right reversal. PMID- 14562092 TI - Kinetics: Gaussian statistics in granular gases. PMID- 14562094 TI - Planning for smallpox outbreaks. AB - Mathematical models of viral transmission and control are important tools for assessing the threat posed by deliberate release of the smallpox virus and the best means of containing an outbreak. Models must balance biological realism against limitations of knowledge, and uncertainties need to be accurately communicated to policy-makers. Smallpox poses the particular challenge that key biological, social and spatial factors affecting disease spread in contemporary populations must be elucidated largely from historical studies undertaken before disease eradication in 1979. We review the use of models in smallpox planning within the broader epidemiological context set by recent outbreaks of both novel and re-emerging pathogens. PMID- 14562095 TI - Global analysis of protein localization in budding yeast. AB - A fundamental goal of cell biology is to define the functions of proteins in the context of compartments that organize them in the cellular environment. Here we describe the construction and analysis of a collection of yeast strains expressing full-length, chromosomally tagged green fluorescent protein fusion proteins. We classify these proteins, representing 75% of the yeast proteome, into 22 distinct subcellular localization categories, and provide localization information for 70% of previously unlocalized proteins. Analysis of this high resolution, high-coverage localization data set in the context of transcriptional, genetic, and protein-protein interaction data helps reveal the logic of transcriptional co-regulation, and provides a comprehensive view of interactions within and between organelles in eukaryotic cells. PMID- 14562096 TI - Three-dimensional magnetic field topology in a region of solar coronal heating. AB - Flares and X-ray jets on the Sun arise in active regions where magnetic flux emerges from the solar interior amd interacts with the ambient magnetic field. The interactions are believed to occur in electric current sheets separating regions of opposite magnetic polarity. The current sheets located in the corona or upper chromosphere have long been thought to act as an important source of coronal heating, requiring their location in the corona or upper chromosphere. The dynamics and energetics of these sheets are governed by a complex magnetic field structure that, until now, has been difficult to measure. Here we report the determination of the full magnetic vector in an interaction region near the base of the solar corona. The observations reveal two magnetic features that characterize young active regions on the Sun: a set of rising magnetic loops and a tangential discontinuity of the magnetic field direction, the latter being the observational signature of an electric current sheet. This provides strong support for coronal heating models based on the dissipation of magnetic energy at current sheets. PMID- 14562097 TI - The speed of information in a 'fast-light' optical medium. AB - One consequence of the special theory of relativity is that no signal can cause an effect outside the source light cone, the space-time surface on which light rays emanate from the source. Violation of this principle of relativistic causality leads to paradoxes, such as that of an effect preceding its cause. Recent experiments on optical pulse propagation in so-called 'fast-light' media- which are characterized by a wave group velocity upsilon(g) exceeding the vacuum speed of light c or taking on negative values--have led to renewed debate about the definition of the information velocity upsilon(i). One view is that upsilon(i) = upsilon(g) (ref. 4), which would violate causality, while another is that upsilon(i) = c in all situations, which would preserve causality. Here we find that the time to detect information propagating through a fast-light medium is slightly longer than the time required to detect the same information travelling through a vacuum, even though upsilon(g) in the medium vastly exceeds c. Our observations are therefore consistent with relativistic causality and help to resolve the controversies surrounding superluminal pulse propagation. PMID- 14562098 TI - Single-electron transistor of a single organic molecule with access to several redox states. AB - A combination of classical Coulomb charging, electronic level spacings, spin, and vibrational modes determines the single-electron transfer reactions through nanoscale systems connected to external electrodes by tunnelling barriers. Coulomb charging effects have been shown to dominate such transport in semiconductor quantum dots, metallic and semiconducting nanoparticles, carbon nanotubes, and single molecules. Recently, transport has been shown to be also influenced by spin--through the Kondo effect--for both nanotubes and single molecules, as well as by vibrational fine structure. Here we describe a single electron transistor where the electronic levels of a single pi-conjugated molecule in several distinct charged states control the transport properties. The molecular electronic levels extracted from the single-electron-transistor measurements are strongly perturbed compared to those of the molecule in solution, leading to a very significant reduction of the gap between the highest occupied molecular orbital and the lowest unoccupied molecular orbital. We suggest, and verify by simple model calculations, that this surprising effect could be caused by image charges generated in the source and drain electrodes resulting in a strong localization of the charges on the molecule. PMID- 14562099 TI - Zero thermal expansion in YbGaGe due to an electronic valence transition. AB - Most materials expand upon heating. Although rare, some materials expand on cooling, and are said to exhibit negative thermal expansion (NTE); but the property is exhibited in only one crystallographic direction. Such materials include silicon and germanium at very low temperature (<100 K) and, at room temperature, glasses in the titania-silica family, Kevlar, carbon fibres, anisotropic Invar Fe-Ni alloys, ZrW2O3 (ref. 4) and certain molecular networks. NTE materials can be combined with materials demonstrating a positive thermal expansion coefficient to fabricate composites exhibiting an overall zero thermal expansion (ZTE). ZTE materials are useful because they do not undergo thermal shock on rapid heating or cooling. The need for such composites could be avoided if ZTE materials were available in a pure form. Here we show that an electrically conductive intermetallic compound, YbGaGe, can exhibit nearly ZTE--that is, negligible volume change between 100 and 400 K. We suggest that this response is due to a temperature-induced valence transition in the Yb atoms. ZTE materials are desirable to prevent or reduce resulting strain or internal stresses in systems subject to large temperature fluctuations, such as in space applications and thermomechanical actuators. PMID- 14562100 TI - Reduction of soil carbon formation by tropospheric ozone under increased carbon dioxide levels. AB - In the Northern Hemisphere, ozone levels in the troposphere have increased by 35 per cent over the past century, with detrimental impacts on forest and agricultural productivity, even when forest productivity has been stimulated by increased carbon dioxide levels. In addition to reducing productivity, increased tropospheric ozone levels could alter terrestrial carbon cycling by lowering the quantity and quality of carbon inputs to soils. However, the influence of elevated ozone levels on soil carbon formation and decomposition are unknown. Here we examine the effects of elevated ozone levels on the formation rates of total and decay-resistant acid-insoluble soil carbon under conditions of elevated carbon dioxide levels in experimental aspen (Populus tremuloides) stands and mixed aspen-birch (Betula papyrifera) stands. With ambient concentrations of ozone and carbon dioxide both raised by 50 per cent, we find that the formation rates of total and acid-insoluble soil carbon are reduced by 50 per cent relative to the amounts entering the soil when the forests were exposed to increased carbon dioxide alone. Our results suggest that, in a world with elevated atmospheric carbon dioxide concentrations, global-scale reductions in plant productivity due to elevated ozone levels will also lower soil carbon formation rates significantly. PMID- 14562101 TI - Flying and swimming animals cruise at a Strouhal number tuned for high power efficiency. AB - Dimensionless numbers are important in biomechanics because their constancy can imply dynamic similarity between systems, despite possible differences in medium or scale. A dimensionless parameter that describes the tail or wing kinematics of swimming and flying animals is the Strouhal number, St = fA/U, which divides stroke frequency (f) and amplitude (A) by forward speed (U). St is known to govern a well-defined series of vortex growth and shedding regimes for airfoils undergoing pitching and heaving motions. Propulsive efficiency is high over a narrow range of St and usually peaks within the interval 0.2 < St < 0.4 (refs 3 8). Because natural selection is likely to tune animals for high propulsive efficiency, we expect it to constrain the range of St that animals use. This seems to be true for dolphins, sharks and bony fish, which swim at 0.2 < St < 0.4. Here we show that birds, bats and insects also converge on the same narrow range of St, but only when cruising. Tuning cruise kinematics to optimize St therefore seems to be a general principle of oscillatory lift-based propulsion. PMID- 14562102 TI - New frog family from India reveals an ancient biogeographical link with the Seychelles. AB - About 96% of the more than 4,800 living anuran species belong to the Neobatrachia or advanced frogs. Because of the extremely poor representation of these animals in the Mesozoic fossil record, hypotheses on their early evolution have to rely largely on extant taxa. Here we report the discovery of a burrowing frog from India that is noticeably distinct from known taxa in all anuran families. Phylogenetic analyses of 2.8 kilobases of mitochondrial and nuclear DNA unambiguously designate this frog as the sister taxon of Sooglossidae, a family exclusively occurring on two granitic islands of the Seychelles archipelago. Furthermore, molecular clock analyses uncover the branch leading to both taxa as an ancient split in the crown-group Neobatrachia. Our discovery discloses a lineage that may have been more diverse on Indo-Madagascar in the Cretaceous period, but now only comprises four species on the Seychelles and a sole survivor in India. Because of its very distinct morphology and an inferred origin that is earlier than several neobatrachian families, we recognize this frog as a new family. PMID- 14562103 TI - Females increase offspring heterozygosity and fitness through extra-pair matings. AB - Females in a variety of species commonly mate with multiple males, and there is evidence that they benefit by producing offspring of higher genetic quality; however, the nature of these genetic benefits is debated. Enhanced offspring survival or quality can result from intrinsic effects of paternal genes---'good genes'--or from interactions between the maternal and paternal genomes- 'compatible genes'. Evidence for the latter process is accumulating: matings between relatives lead to decreased reproductive success, and the individual level of inbreeding--measured as average heterozygosity--is a strong fitness predictor. Females should thus benefit from mating with genetically dissimilar males. In many birds, social monogamy restricts mate choice, but females may circumvent this by pursuing extra-pair copulations. Here we show that female blue tits, Parus caeruleus, increase the heterozygosity of their progeny through extra pair matings. Females thereby produce offspring of higher reproductive value, because less inbred individuals have increased survival chances, a more elaborate male secondary sexual trait (crown colour) and higher reproductive success. The cost of inbreeding may therefore be an important factor driving the evolution of female extra-pair mating. PMID- 14562104 TI - RNA molecules stimulate prion protein conversion. AB - Much evidence supports the hypothesis that the infectious agents of prion diseases are devoid of nucleic acid, and instead are composed of a specific infectious protein. This protein, PrP(Sc), seems to be generated by template induced conformational change of a normally expressed glycoprotein, PrP(C) (ref. 2). Although numerous studies have established the conversion of PrP(C) to PrP(Sc) as the central pathogenic event of prion disease, it is unknown whether cellular factors other than PrP(C) might be required to stimulate efficient PrP(Sc) production. We investigated the biochemical amplification of protease resistant PrP(Sc)-like protein (PrPres) using a modified version of the protein misfolding cyclic amplification method. Here we report that stoichiometric transformation of PrP(C) to PrPres in vitro requires specific RNA molecules. Notably, whereas mammalian RNA preparations stimulate in vitro amplification of PrPres, RNA preparations from invertebrate species do not. Our findings suggest that host-encoded stimulatory RNA molecules may have a role in the pathogenesis of prion disease. They also provide a practical approach to improve the sensitivity of diagnostic techniques based on PrPres amplification. PMID- 14562105 TI - Functional cloning of TUG as a regulator of GLUT4 glucose transporter trafficking. AB - Insulin stimulates glucose uptake in fat and muscle by mobilizing the GLUT4 glucose transporter. GLUT4 is sequestered intracellularly in the absence of insulin, and is redistributed to the plasma membrane within minutes of insulin stimulation. But the trafficking mechanisms that control GLUT4 sequestration have remained elusive. Here we describe a functional screen to identify proteins that modulate GLUT4 distribution, and identify TUG as a putative tether, containing a UBX domain, for GLUT4. In truncated form, TUG acts in a dominant-negative manner to inhibit insulin-stimulated GLUT4 redistribution in Chinese hamster ovary cells and 3T3-L1 adipocytes. Full-length TUG forms a complex specifically with GLUT4; in 3T3-L1 adipocytes, this complex is present in unstimulated cells and is largely disassembled by insulin. Endogenous TUG is localized with the insulin mobilizable pool of GLUT4 in unstimulated 3T3-L1 adipocytes, and is not mobilized to the plasma membrane by insulin. Distinct regions of TUG are required to bind GLUT4 and to retain GLUT4 intracellularly in transfected, non-adipose cells. Our data suggest that TUG traps endocytosed GLUT4 and tethers it intracellularly, and that insulin mobilizes this pool of retained GLUT4 by releasing this tether. PMID- 14562106 TI - Global analysis of protein expression in yeast. AB - The availability of complete genomic sequences and technologies that allow comprehensive analysis of global expression profiles of messenger RNA have greatly expanded our ability to monitor the internal state of a cell. Yet biological systems ultimately need to be explained in terms of the activity, regulation and modification of proteins--and the ubiquitous occurrence of post transcriptional regulation makes mRNA an imperfect proxy for such information. To facilitate global protein analyses, we have created a Saccharomyces cerevisiae fusion library where each open reading frame is tagged with a high-affinity epitope and expressed from its natural chromosomal location. Through immunodetection of the common tag, we obtain a census of proteins expressed during log-phase growth and measurements of their absolute levels. We find that about 80% of the proteome is expressed during normal growth conditions, and, using additional sequence information, we systematically identify misannotated genes. The abundance of proteins ranges from fewer than 50 to more than 10(6) molecules per cell. Many of these molecules, including essential proteins and most transcription factors, are present at levels that are not readily detectable by other proteomic techniques nor predictable by mRNA levels or codon bias measurements. PMID- 14562109 TI - It could be worse. PMID- 14562110 TI - Singapore: filling biopolis. PMID- 14562111 TI - Expression of FLIP(Long) and FLIP(Short) in bone marrow mononuclear and CD34+ cells in patients with myelodysplastic syndrome: correlation with apoptosis. AB - Several apoptosis-inducing systems, including Fas/Fas ligand and TNF-related apoptosis-inducing ligand (TRAIL) and its receptors, are upregulated in myelodysplastic syndrome (MDS). FLIP (FLICE (FAS-associated death-domain-like IL 1beta-converting enzyme)-inhibitory protein)) was identified as an inhibitor of FAS and TRAIL signals. Here, we characterized FLIP(Long) (FLIP(L)) and FLIP(Short) (FLIP(S)) expression in bone marrow mononuclear cells (BMMNCs) and in CD34+ cells of 29 MDS patients, and in 17 normal volunteers. The expression was correlated with apoptotic indices. In CD34+ cells, FLIP(L) levels were higher among normal individuals than in MDS patients (P=0.04). Among total BMMNC, FLIP(L) levels also tended to be higher in normal subjects than in MDS patients, although this difference was not significant (P=0.71). FLIP(L) levels in CD34+ cells were negatively correlated with apoptosis in both normal and MDS marrows (P=0.03). FLIP(Short) RNA expression was higher in MDS patients than in normal controls in both BMMNC (P=0.03) and CD34+ cells (P=0.08). In contrast to FLIP(L), FLIP(St) levels were positively correlated with apoptosis. At the protein level FLIP was most readily detectable in patients with high blast counts. The data suggest that FLIP(L) and FLIP(S) are differentially regulated, and that the relative levels of both isoforms play a role in the regulation of apoptosis in MDS. PMID- 14562112 TI - Heterogeneous copy numbers of API2-MALT1 chimeric transcripts in mucosa associated lymphoid tissue lymphoma. AB - T(11;18)(q21;q21) results in a chimeric transcript between API2 at 11q21 and MALT1 at 18q21 and is a characteristic chromosomal aberration of extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma). API2-MALT1 chimeric transcripts are present in approximately one-third of all cases of MALT lymphoma. MALT lymphoma is also known to have variations in histological features and tumor cell proportions. Real-time polymerase chain reaction (PCR) was used to examine number of API2-MALT1 copies in clinical samples for further investigation of the pathophysiology of MALT lymphoma. A total of 13 samples of MALT lymphoma contained API2-MALT1 transcripts from 1.7 x 10(-2) to 1.0 copies/beta-actin copy. These findings were compared to the proportions of tumor cells in genomic VDJ PCR products determined by Southern blotting. Tumor cell ratios varied widely among the patients' samples, and no significant correlation was found between transcript copy number and tumor cell ratio. These results suggest that copy numbers of API2-MALT1 do not reflect tumor cell proportions, and that the number of copies of API2-MALT1 in a tumor cell is different for each clinical sample. PMID- 14562113 TI - B-cell development in the presence of the MLL/AF4 oncoprotein proceeds in the absence of HOX A7 and HOX A9 expression. AB - Infant acute lymphoblastic leukemia (ALL) is frequently characterized by the t(4;11)(q21;q23) cytogenetic abnormality encoding the MLL/AF4 oncogene, increased HOX gene expression and a pro-B/monocytoid phenotype. We have previously established a novel MLL/AF4-positive cell line, B-lineage 3 (BLIN-3), which retains several features of normal B-lineage development (functional Ig gene rearrangement and apoptotic sensitivity to stromal cell withdrawal) not generally observed in infant ALL. We now use microarray analysis to identify patterns of gene expression in BLIN-3 that may modulate MLL/AF4 oncogenesis and contribute to the retention of normal B-lineage developmental characteristics. Comparison of 6815 expressed genes in BLIN-3 with published microarray data on leukemic blasts from t(4;11) patients indicated that BLIN-3 was unique in lacking the expression of certain HOX-A cluster genes. These results were validated by RT-PCR showing no expression of HOX A7 or HOX A9 in BLIN-3. A HOX C8 promoter reporter was active in BLIN-3, indicating that lack of HOX gene expression in BLIN-3 was not due to a nonfunctional MLL/AF4. Our results suggest that B-lineage development can proceed in t(4;11) leukemic blasts in the absence of HOX-A gene expression. PMID- 14562114 TI - Telomerase is limiting the growth of acute myeloid leukemia cells. AB - Telomeres play an important role in the proliferation and senescence of normal and malignant cells. To test the role of telomerase in acute myeloid leukemia (AML), we expressed the telomerase reverse transcriptase (hTERT) gene, a dominant negative hTERT (DN-hTERT) (D868A, D869A) gene, or a gene encoding green fluorescence protein (GFP) in the leukemia cell line K562 and in primary AML cells from different patients, using retroviral vectors. Cells transduced with hTERT exhibited elevated levels of telomerase activity compared to GFP controls, whereas cells expressing DN-hTERT had decreased telomerase activity. K562 populations transduced with DN-hTERT showed reduced clonogenicity, telomere dysfunction and increased numbers of apoptotic cells compared to GFP- or hTERT transduced cells. Two of four clones transduced with DN-hTERT died after 30 and 53 population doublings, respectively. Transduced AML cells were tested in primary colony-forming unit (CFU) and suspension culture assays. Relative to hTERT- and GFP-transduced controls, AML cells transfected with DN-hTERT produced fewer CFU and showed lower engraftment after transplantation into sublethally irradiated beta(2)-m(-/-) nonobese diabetic/severe combined immunodeficient mice. We conclude that telomerase is limiting the growth of the leukemic cell line K562 and primary AML progenitor cells. Our data warrant further studies of the therapeutic use of telomerase inhibitors in AML. PMID- 14562115 TI - Stimulation of non-Hodgkin's lymphoma via HVEM: an alternate and safe way to increase Fas-induced apoptosis and improve tumor immunogenicity. AB - Stimulation by CD40 ligand (L) improves B-cell malignancy immunogenicity, and also induces proliferative signals. To avoid these tumorigenic effects, we studied an alternate way of tumor-cell stimulation by homologous to lymphotoxin, inducible expression, competing for GpD of herpesvirus, which binds to the herpesvirus entry mediator (HVEM), and is expressed on T-lymphocytes (LIGHT), the ligand for HVEM, a new member of the tumor necrosis factor (TNF)/TNF-receptor ( R) family. HVEM is constitutively expressed on the surface of tumor B cells. We focused our attention on mantle cell lymphoma, a subtype of B-cell malignancy of poor prognosis. Triggering by LIGHT, in contrast to CD40L stimulation, did not increase lymphoma proliferation nor decrease chemotherapy entrance. We observed an upregulation of the TNFR apoptosis-inducing ligand Fas, and in contrast to CD40L-induced protection, an enhancement of lymphoma sensitivity to Fas-induced apoptosis. LIGHT triggering increased lymphoma cell recognition in a mixed lymphocyte response. In conclusion, LIGHT-mediated triggering renders B-cell lymphomas more immunogenic and sensitive to apoptosis, without inducing proliferation. Since LIGHT triggering also enhances the functions of T lymphocytes and dendritic cells, it could be a unique way to restore an efficient cancer control by its pleiotropic effects on immune effectors and tumor cells. PMID- 14562116 TI - Synaptojanin 2 is recognized by HLA class II-restricted hairy cell leukemia specific T cells. AB - Hairy cell leukemia (HCL) is a chronic mature B-cell leukemia characterized by malignant B cells that have typical hairy protrusions. To characterize possible HCL-associated tumor antigens, we generated an HCL-specific and HLA class II (DPw4)-restricted proliferative CD4+ T-cell clone. To identify the target antigen of these T cells, we constructed a synthetic peptide library dedicated to bind HLA DPw4, and identified a mimicry epitope recognized by the T-cell clone. With this epitope, the recognition motif of the T-cell clone was deduced and a peptide of human synaptojanin 2 (Syn 2) was identified that stimulated the HCL-reactive T cell clone. Both Northern and Western blot analyses showed that Syn 2 expression was increased in HCL samples compared to other B cells. Besides, the Syn 2 expressing cell line AML193, with the introduced restrictive HLA-DPw4 molecules, was recognized by the HCL-specific T-cell clone. These results indicate that Syn 2 is a target of autoreactive HCL-specific T cells. Since Syn 2 is a phosphatidylinositol 4,5-biphosphatase involved in cell growth and rearrangement of actin filaments, the increased Syn 2 expression may correlate with the disease etiology or the characteristic morphologic alterations caused by the disease. PMID- 14562117 TI - Marrow fibrosis, indicator of therapy failure in chronic myeloid leukemia - prospective long-term results from a randomized-controlled trial. AB - Marrow fibrosis (MF) has rarely been considered in therapy studies on chronic myeloid leukemia (CML), and there is a lack of long-term observations on the basis of sequential bone marrow biopsies (BMBs) taken prospectively during the course of disease. A total of 848 BMBs from 400 patients with Ph(+) CML recruited in the German randomized CML study I were examined for MF before and during therapy. In total, 110 patients had been randomized to receive interferon (IFN) alpha, and 290 to receive chemotherapy (hydroxyurea (HU): 154, busulfan: 136). During IFN-alpha and HU medication, MF was reduced or did not increase for about 2 years. Evolving or progressive MF was an independent and early predictor of therapy failure about 2 years earlier than indicated by changes in the peripheral blood, spleen size, marrow blast count and cytogenetics (P<0.00005), resulting in a significant shortening of the survival times of patients independent of the type of therapy applied including allografting (multivariate analyses; P<0.00005). The analyzed long-term observations strongly indicate that MF is an independent poor prognostic complication of CML, allowing an early prediction of therapy failure. Consideration of the fiber content in marrow may therefore significantly improve the prediction of therapy efficacy and outcome of disease. PMID- 14562118 TI - Autologous peripheral blood stem cell transplantation of stem cells harvested in imatinib-induced complete cytogenetic remission: an example of in vivo purging in CML. PMID- 14562119 TI - Dual mutations in the AML1 and FLT3 genes are associated with leukemogenesis in acute myeloblastic leukemia of the M0 subtype. AB - Point mutations of the transcription factor AML1 are associated with leukemogenesis in acute myeloblastic leukemia (AML). Internal tandem duplications (ITDs) in the juxtamembrane domain and mutations in the second tyrosine kinase domain of the Fms-like tyrosine kinase 3 (FLT3) gene represent the most frequent genetic alterations in AML. However, such mutations per se appear to be insufficient for leukemic transformation. To evaluate whether both AML1 and FLT3 mutations contribute to leukemogenesis, we analyzed mutations of these genes in AML M0 subtype in whom AML1 mutations were predominantly observed. Of 51 patients, eight showed a mutation in the Runt domain of the AML1 gene: one heterozygous missense mutation with normal function, five heterozygous frameshift mutations and two biallelic nonsense or frameshift mutations, resulting in haploinsufficiency or complete loss of the AML1 activities. On the other hand, a total of 10 of 49 patients examined had the FLT3 mutation. We detected the FLT3 mutation in five of eight (63%) patients with AML1 mutation, whereas five of 41 (12%) without AML1 mutation showed the FLT3 mutation (P=0.0055). These observations suggest that reduced AML1 activities predispose cells to the acquisition of the activating FLT3 mutation as a secondary event leading to full transformation in AML M0. PMID- 14562120 TI - Treatment by design in leukemia, a meeting report, Philadelphia, Pennsylvania, December 2002. AB - Novel approaches have been designed to treat leukemia based on our understanding of the genetic and biochemical lesions present in different malignancies. This meeting report summarizes some of the recent advances in leukemia treatment. Based on the discoveries of cellular oncogenes, chromosomal translocations, monoclonal antibodies, multidrug resistance pumps, signal transduction pathways, genomics/proteonomic approaches to clinical diagnosis and mutations in biochemical pathways, clinicians and basic scientists have been able to identify the particular genetic mutations and signal transduction pathways involved as well as design more appropriate treatments for the leukemia patient. This meeting report discusses these exciting new therapies and the results obtained from ongoing clinical trials. Furthermore, rational approaches to treat complications of tumor lysis syndrome by administration of the recombinant urate oxidase protein, also known as rasburicase, which corrects the biochemical defect present in humans, were discussed. Clearly, over the past 25 years, molecular biology and biotechnology has provided the hematologist/oncologist novel bullets in their arsenal that will allow treatment by design in leukemia. PMID- 14562121 TI - Increased expression of fibroblast growth factor receptor 3 in CD34+ BCR-ABL+ cells from patients with chronic myeloid leukemia. AB - Previously, we showed that expression of myeloma-associated (proto)oncogene fibroblast growth factor receptor 3 (FGFR-3) is increased in white blood cells from patients with chronic myeloid leukemia (CML). The abnormal expression was returned back to the normal levels as soon as these patients reconstituted their hematopoiesis following transplantation of allogeneic peripheral blood stem cells. The aims of this study were: (1) to define population(s) of cells overexpressing FGFR-3, and (2) to determine the expression of FGFR-3 during the clinical course of the disease. We show that the vast majority of FGFR-3 transcripts as well as FGFR-3 protein arise from CD34+ BCR-ABL+ cells. Although increased levels of FGFR-3 were found in majority of late chronic phase patients treated with interferon alpha or hydroxyurea, the expression of FGFR-3 was always lowered following treatment with BCR-ABL tyrosine kinase inhibitor STI571. Compared to unstimulated cells, high levels of FGFR-3 were also identified in CD34+ cells from granulocyte colony-stimulating factor-mobilized blood stem cell harvests from healthy donors, suggesting a potential growth factor-dependent basis for elevated expression of FGFR-3 in CML. These findings have implications for the involvement of FGFR-3 in malignant hematopoiesis and depict FGFR-3 tyrosine kinase in CD34+ leukemic cells as a possible target for tyrosine kinase inhibitors. PMID- 14562122 TI - A cryptic insertion (17;15) on both chromosomes 17 with lack of PML-RARA expression in a case of atypical acute promyelocytic leukemia. PMID- 14562123 TI - A further milestone towards comprehensive standardization of quantitative RT-PCR protocols for leukemic fusion gene transcripts has been reached. PMID- 14562124 TI - Evaluation of candidate control genes for diagnosis and residual disease detection in leukemic patients using 'real-time' quantitative reverse transcriptase polymerase chain reaction (RQ-PCR) - a Europe against cancer program. AB - Real-time quantitative RT-PCR (RQ-PCR) is a sensitive tool to monitor minimal residual disease (MRD) in leukemic patients through the amplification of a fusion gene (FG) transcript. In order to correct variations in RNA quality and quantity and to calculate the sensitivity of each measurement, a control gene (CG) transcript should be amplified in parallel to the FG transcript. To identify suitable CGs, a study group within the Europe Against Cancer (EAC) program initially focused on 14 potential CGs using a standardized RQ-PCR protocol. Based on the absence of pseudogenes and the level and stability of the CG expression, three genes were finally selected: Abelson (ABL), beta-2-microglobulin (B2M), and beta-glucuronidase (GUS). A multicenter prospective study on normal (n=126) and diagnostic leukemic (n=184) samples processed the same day has established reference values for the CG expression. A multicenter retrospective study on over 250 acute and chronic leukemia samples obtained at diagnosis and with an identified FG transcript confirmed that the three CGs had a stable expression in the different types of samples. However, only ABL gene transcript expression did not differ significantly between normal and leukemic samples at diagnosis. We therefore propose to use the ABL gene as CG for RQ-PCR-based diagnosis and MRD detection in leukemic patients. Overall, these data are not only eligible for quantification of fusion gene transcripts, but also for the quantification of aberrantly expressed genes. PMID- 14562126 TI - Increased serum levels of interleukin-9 correlate to negative prognostic factors in Hodgkin's lymphoma. AB - Hodgkin's lymphoma (HL) is characterised by an unbalanced cytokine secretion. Many of these cytokines have been implicated in the regulation of malignant and infiltrating cells. Interleukin-9 (IL-9) has been described to act in an autocrine fashion in HL, stimulating proliferation of the malignant cells. To investigate the potential clinical implication of this observation, a novel ELISA method was used to examine the serum levels of IL-9 in lymphoma patients. High levels of IL-9 were found in the sera from patients with HL (18/44), but not in the sera from non-Hodgkin's lymphoma patients (3/21) or healthy controls. The highest serum IL-9 levels, up to 3350 pg/ml, were observed in the nodular sclerosis subtype, and there was a correlation between IL-9 levels and the negative prognostic factors advanced stage, B-symptoms, low blood Hb and high erythrocyte sedimentation rate. Furthermore, there was no correlation between serum levels of IL-9 and IL-13, a cytokine where serum levels have been speculated to be of clinical importance. This is the first report showing that IL 9 can be measured in serum samples. A novel correlation between increased serum IL-9 levels, HL and clinical features is shown, suggesting that IL-9 is a candidate factor contributing to the development of HL. PMID- 14562125 TI - Standardization and quality control studies of 'real-time' quantitative reverse transcriptase polymerase chain reaction of fusion gene transcripts for residual disease detection in leukemia - a Europe Against Cancer program. AB - Detection of minimal residual disease (MRD) has proven to provide independent prognostic information for treatment stratification in several types of leukemias such as childhood acute lymphoblastic leukemia (ALL), chronic myeloid leukemia (CML) and acute promyelocytic leukemia. This report focuses on the accurate quantitative measurement of fusion gene (FG) transcripts as can be applied in 35 45% of ALL and acute myeloid leukemia, and in more than 90% of CML. A total of 26 European university laboratories from 10 countries have collaborated to establish a standardized protocol for TaqMan-based real-time quantitative PCR (RQ-PCR) analysis of the main leukemia-associated FGs within the Europe Against Cancer (EAC) program. Four phases were scheduled: (1) training, (2) optimization, (3) sensitivity testing and (4) patient sample testing. During our program, three quality control rounds on a large series of coded RNA samples were performed including a balanced randomized assay, which enabled final validation of the EAC primer and probe sets. The expression level of the nine major FG transcripts in a large series of stored diagnostic leukemia samples (n=278) was evaluated. After normalization, no statistically significant difference in expression level was observed between bone marrow and peripheral blood on paired samples at diagnosis. However, RQ-PCR revealed marked differences in FG expression between transcripts in leukemic samples at diagnosis that could account for differential assay sensitivity. The development of standardized protocols for RQ-PCR analysis of FG transcripts provides a milestone for molecular determination of MRD levels. This is likely to prove invaluable to the management of patients entered into multicenter therapeutic trials. PMID- 14562127 TI - Comparison between TaqMan and LightCycler technologies for quantification of minimal residual disease by using immunoglobulin and T-cell receptor genes consensus probes. AB - Quantification of residual leukemic cells at early time points during therapy can reliably predict the outcome in children with acute lymphoblastic leukemia (ALL). Recently, semiquantitative minimal residual disease (MRD) detection assays such as dot-blot hybridization have been replaced by real-time quantitative PCR. We tested the flexibility of the two most used real-time PCR machines: the SDS 7700 or 'TaqMan' (TM) (Applied Biosystems) and the LightCycler (LC) (Roche) instruments. Clonal T-cell receptor and immunoglobulin gene rearrangements were used for MRD detection with germline hydrolyzation probes and clone-specific primers. Sensitivity tests for 65 clonal gene rearrangements and MRD quantification in 90 bone marrow samples during therapy of 49 children with ALL at diagnosis or relapse were performed with both machines. Both real-time PCR systems provided specific results for MRD quantification in all follow-up samples. In conclusion, we were able to demonstrate that TM and LC real-time PCR technologies produce similar MRD quantification results and that the quantification assays can be easily transferred from one detection system to the other. Using the same detection format, both techniques can be applied in combination in multicenter MRD studies. PMID- 14562128 TI - Premature to early ejaculation: a sampling of manuscripts regarding the most common male sexual dysfunction published in the IJIR: The Journal of Sexual Medicine. PMID- 14562129 TI - Towards evidence-based drug treatment research on premature ejaculation: a critical evaluation of methodology. AB - In the last decade, an increasing number of drug studies on premature ejaculation have been published. The methodology used in these studies differed widely. In this review, it is therefore questioned as to how far the differences in methodology could have influenced clinical outcomes. The aim of the present review was to compare the different methodologies that were used in drug studies on premature ejaculation. The majority of these studies were conducted between 1973 and 2003 and were strikingly different in study design and in the quantification of clinical outcomes. It appeared that in some of these studies, placebo and active drug effects were neglected due to an erroneous methodology. The few studies that were using stopwatch assessment at each intercourse at baseline and during treatment and in which the intravaginal ejaculation latency time (IELT) was included appeared to be comparable in their results. The use of a well-defined definition of premature ejaculation, for example, an IELT of less than 1 min revealed reproducible results. Finally, the retrospective use of a questionnaire or a subjective report on ejaculation time induced higher effects with regard to placebo effects and an underestimation of active drug effects. In conclusion, for drug treatment research of premature ejaculation it is recommended to use a randomized double-blind prospective design, the use of the IELT, the use of a stopwatch at each coitus both during a baseline period and during drug treatment and a definition of premature ejaculation as an ejaculation that occurs within 1 min after vaginal penetration. PMID- 14562130 TI - Antihypertensive treatment and erectile dysfunction in a cohort of type II diabetes patients. AB - The association between different antihypertensive drugs and erectile dysfunction (ED) was examined in a cohort of type II diabetes patients identified in the UK General Practice Research Database (GPRD). The GPRD contains details of diagnoses, prescribing, investigations, risk factors, outcomes, and hospital referrals, together with basic demographic information for approximately six million patients from more than 450 representative general practices throughout the UK. A total of 634 cases and 2526 controls were included for analysis. Unconditional logistic regression analysis was performed to assess the risk of ED after adjusting for age at diabetes diagnosis date, cigarette smoking, depression, glycemic control, use of HMG-CoA reductase inhibitors, use of histamine receptor antagonists, use of digitalis medicines, and use of nitrates. Increased risk of ED was observed among patients taking the following antihypertensives: ACE inhibitors (OR=1.47, 95% CI=1.21, 1.80) and alpha blockers (OR=1.54, 95% CI=1.11, 2.12). However, we identified a nearly 30% reduction in risk among patients on diuretics (OR=0.73, 95% CI=0.54, 0.99). No statistically significant increase in risk was observed among users of beta blockers and calcium channel blockers (OR=1.05, 95% CI=0.85, 1.31) and (OR=1.14, 95% CI=0.91, 1.43), respectively. The results of this study confirm the strong and recognized effect of comorbidities in a diabetic population, but also require additional experimental and observational studies to further understand the potential benefit of diuretics and other ED treatments such as PDE5 inhibitors. PMID- 14562131 TI - Long-term efficacy and compliance of intracorporeal (IC) injection for erectile dysfunction following radical prostatectomy: SHIM (IIEF-5) analysis. AB - Baseline and follow-up data from 102 patients using intracorporeal (IC) injection for erectile dysfunction (ED) following RP were retrospectively collected. We compared baseline International Index for Erectile Function (IIEF) questionnaires with the abridged IIEF-5 questionnaires, referred to as the Sexual Health Inventory of Men (SHIM) to determine drug efficacy. The mean presurgery SHIM score was 21.75+/-5.23, which decreased to 4.23+/-3.48 after surgery and increased to 19.46+/-8.78 post-treatment. Overall, 68% (69/102) of patients achieved and maintained erections sufficient for sexual intercourse and 48% (49/102) of patients continued long-term therapy with a mean use of 3.7+/-1.9 y. In all, 52% (53/102) patients discontinued IC therapy. However when excluding patients who switched to oral therapy, had loss of partner or return of normal erections; the compliance to IC injections was 70.6% (71/102). There was no difference in the SHIM analysis between the nerve sparing (NS) and the non-NS or between the types of medications used. IC injections can provide excellent long term efficacy and compliance in up to 70% of the patients. This study suggests that IC injections are an excellent salvage option in NS patients who fail oral therapy and a first option in patients with non-NS procedures. PMID- 14562132 TI - Prevalence and risk factors for erectile dysfunction in primary care: results of a Korean study. AB - In order to assess the prevalence and associated factors for erectile dysfunction (ED) in primary care, a cross-sectional study was undertaken by questionnaire distributed to consecutive adult male attendees at 32 family practices. ED was assessed by the Korean five-item version of the International Index of Erectile Function (IIEF-5). In total, 3501 completed questionnaires were available for analysis. The prevalence of ED was severe (IIEF-5 score: 5-9) in 1.6% of cases, moderate (10-13) in 10.2%, mild (14-17) in 24.7%, and normal (18-25) in 63.4%. The prevalence of ED increased with age, lower educational status, heavy job related physical activity, and lower income. ED prevalence was significantly higher in patients with chronic diseases such as diabetes, depression, and anxiety. These results suggest that the age-adjusted prevalence of ED among Korean men can be estimated as 32.2% (95% CI 30.6-33.7). Low socioeconomic status and several diseases such as diabetes, anxiety, and depression, as well as age, were associated with ED. PMID- 14562133 TI - Cross-cultural adaptation and validation of the English version of the International Index of Erectile Function (IIEF) for use in Malaysia. AB - We adapted the English International Index of Erectile Function (IIEF) into Malay. This was difficult as many sex-related terms do not exist in colloquial Malay. In the pretest, there was no difficulty with comprehension and all subjects judged the Malay IIEF equivalent to the English IIEF. After slight modification, a final instrument was evaluated in two studies. Study A included 136 subjects. It showed that the instrument had good reliability and discriminant validity. The factor structure of the English IIEF was not reproducible. Study B included 26 ED subjects who underwent oral sildenafil therapy. The Malay IIEF was sensitive to treatment response. The area under the ROC curve of the Malay IIEF-5 was 0.86; the optimal cutoff score has a sensitivity of 85% and specificity of 75%. The results suggest that the Malay IIEF requires more work, but the Malay IIEF-5 has acceptable measurement properties to recommend its use in clinical practice and research. PMID- 14562134 TI - Protective role of the glans penis during coitus. AB - To examine the hypothesis that the glans penis acts protectively, absorbing forces, during coitus. Five potent patients (mean age 46.8+/-9.7 y), who had indication for surgical excision of the glans for penile carcinoma were included in the present study. Intraoperatively, intracavernosal pressure (ICP) was adjusted by saline infusion and maintained by a pressure feedback infusion pump to a pressure value of 70 mmHg. Using a dynamometer, an external compressive force of 0.5 kg was applied at the glans penis and the changes in ICP were monitored. Measurements were repeated after surgical excision of the glans. Significant ICP changes were noticed in all patients after excision of the glans. Mean preoperative ICP was 161+/-11.5 mmHg, while after glansectomy it reached 206.6+/-13 mmHg. DeltaICP was 45.8+/-10.57 mmHg. Two of the patients' partners reported pain during intercourse postoperatively, possibly due to the impact of the force applied by the rigid corpora cavernosa on the anterior vaginal wall without any absorption by the glans. The glans penis restricts the increase in ICP during sexual intercourse, playing a protective role for both the corpora cavernosa and the female genitalia. PMID- 14562135 TI - Acupuncture in the treatment of psychogenic erectile dysfunction: first results of a prospective randomized placebo-controlled study. AB - In a prospective study, we investigated the potentially curative effect of acupuncture in patients with psychogenic erectile dysfunction (pED). A total of 22 patients with pED were randomized into two groups. They were either treated with acupuncture specific against ED (treatment group) or acupuncture specific against headache (placebo group). Nonresponders of the placebo group were crossed over to the treatment group. Prior to acupuncture, serum sexual hormone levels, IIEF score, nocturnal penile tumescence testing for three nights (Rigiscan) and the erectile response to 50 mg sildenafil were evaluated. Out of 21 patients, 20 completed the study, including 10 patients after crossover. A satisfactory response was achieved in 68.4% of the treatment group and in 9% of the placebo group (P=0.0017). Another 21.05% of the patients had improved erections, that is, sufficient rigidity under simultaneous treatment with 50 gm sildenafil. The results of our pilot study indicate that acupuncture can be an effective treatment option in more than two-thirds of patients with psychogenic erectile dysfunction. PMID- 14562136 TI - Effect of PDE5 inhibition combined with free oxygen radical scavenger therapy on erectile function in a diabetic animal model. AB - Phosphodiesterase (PDE) inhibitors represent an important advance in the treatment of erectile dysfunction (ED). In spite of widespread use and generally good efficacy, as a class they remain ineffective in 15-57% of men. Specific cohorts of patients with severe vascular or neurogenic basis to their ED, such as diabetic men or those who have undergone radical pelvic surgery, demonstrate lower response rates with PDE inhibition treatment. We believe that circulating levels of nitric oxide (NO) may be enhanced through delivery of adequate concentrations of free oxygen radical scavenger molecules such as vitamin E. Higher levels of NO, theoretically, should produce increased penile blood flow with the potential for a synergistic effect when combined with a PDE5 inhibitor. With this hypothesis in mind, 20 adult male Sprague-Dawley streptozotocin-induced (60 mg/kg i.p.) diabetic rats were divided into four therapeutic groups (n=5). Group I--control animals received peanut oil, group II--vitamin E 20 IU/day, group III--sildenafil 5 mg/kg/day and group IV--vitamin E 20 IU/day plus sildenafil 5 mg/kg/day, by oral gavage daily for 3 weeks. Erectile function was assessed as a rise in intracavernous pressure following cavernous nerve electrostimulation. Penile tissue was harvested to determine the changes in tissue morphology including neuronal nitric oxide synthase, smooth muscle alpha actin and endothelial cell integrity. PDE5 protein content and activity were measured. Significant increases in intracavernous pressure were measured in the animals receiving combined vitamin E plus sildenafil treatment. Immunohistochemical staining showed increases of neuronal nitric oxide synthase, endothelial cell and smooth muscle cell staining. Western blot analysis did not show significant differences of PDE5 protein between the groups. However, higher PDE5 activity was measured in the sildenafil group and lower activity of PDE5 was recorded in the cohort receiving vitamin E with sildenafil. Vitamin E enhanced the therapeutic effect of the PDE5 inhibitor in a meaningful way in this animal model of diabetes. This study indicates a potential means of salvaging erectile function among patients who are refractory to sildenafil. PMID- 14562137 TI - Role of the nitric oxide-cyclic GMP pathway in regulation of vaginal blood flow. AB - The regulatory role of nitric oxide (NO) in vaginal perfusion remains unclear. We used specific inhibitors of enzymes in the NO-cyclic GMP (NO-cGMP) pathway and investigated their effects on vaginal blood flow in the rabbit. NO synthase (NOS) activity was similar in both the proximal and distal rabbit vagina; whereas, arginase activity was 3.4-fold higher in the distal vagina. Intravenous administration of the NOS inhibitor L-NAME resulted in a 66% reduction in genital tissue oxyhemoglobin and a 53% reduction in vaginal blood flow. This attenuation occurred despite a 20-30% increase in systemic arterial pressure. The arginase inhibitor ABH caused a 2.1-fold increase in genital tissue oxyhemoglobin and 34% increase in vaginal blood flow. The guanylate cyclase inhibitor 1H [1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one and the phosphodiesterase type 5 inhibitor sildenafil caused in a 37% reduction and a 44% increase in vaginal blood flow, respectively. These observations suggest that the NO-cGMP pathway is an important regulator of vaginal hemodynamics. PMID- 14562138 TI - Functional prostaglandin E (EP) receptors in human penile corpus cavernosum. AB - In this study, we have characterized functional EP receptors in human corpus cavernosum (HCC) tissue and in HCC smooth muscle cells (SMC). Using RNase protection assays, we determined expression of EP2, EP3I and EP3II receptor mRNA. In organ bath preparations of HCC tissue strips, PGE1 caused dose-dependent relaxation at concentrations below 300 nM. At concentrations greater than 300 nM, PGE1 caused contraction. Addition of the EP1/EP2/EP3 receptor antagonist AH6809 inhibited this contraction and facilitated further relaxation through concentrations above 1 microM of PGE1. The EP1/EP3 receptor selective agonist 17 phenyltrinor-PGE2 caused dose-dependent contraction that was partially attenuated by SC51322, an EP1 selective antagonist. In cultures of HCC SMC, PGE1 stimulated cAMP accumulation in a dose-dependent manner. Interestingly, AH6809 significantly attenuated PGE1-induced cAMP accumulation. Sulprostone, a selective EP3 receptor agonist, induced weak contractions in HCC tissue strips but augmented forskolin induced cAMP synthesis in HCC SMC. The data in this study suggest that HCC and cultured smooth muscle cells express EP1, EP2 and EP3 receptors. These receptors mediate their responses via different biochemical pathways and are expected to have different responses in regulating smooth muscle tone. Thus, we suggest that the ultimate response in erectile tissue to various prostanoids is the integration of responses elicited by individual EP receptor subtypes to a given ligand. PMID- 14562139 TI - Single step isolation of sildenafil from commercially available Viagra tablets. AB - Sildenafil, the active ingredient in Viagra, has been purified from commercially available tablets. The purification, using Sephadex G25 chromatography under conditions of low ionic strength, is simple and inexpensive. Sildenafil purified according to this protocol has been characterized with respect to its IC50 for PDE5, its ultraviolet absorption profile, and by collision-induced dissociation fingerprinting, positive ion nanospray, and MALDI mass spectrometry. Tritated sildenafil (6 Ci/mmol) was prepared commercially using the sildenafil purified by this protocol and was verified to retain the potency of unlabeled sildenafil. This protocol and similar procedures will allow investigators to easily isolate sufficient amounts of sildenafil or other PDE5 inhibitors for conducting biochemical and in vitro studies of drug action. PMID- 14562140 TI - Hyperprolactinemia and sexual function in men: a short review. AB - Erectile dysfunction (ED), generally associated with reduced sexual desire and sometimes with orgasmic or ejaculatory dysfunction, is the major revealing symptom of hyperprolactinemia (HPRL) in men, a condition that should not be neglected since many cases result from pituitary tumors, likely to result in serious complications. It is generally believed that the mechanism of the prolactin (PRL)-induced sexual dysfunctions is a decrease in testosterone secretion. In fact, serum testosterone is normal in many hyperprolactinemic males and there are also testosterone-independent mechanisms, probably mainly set at the level of the brain's neurotransmitter systems. Systematic determinations of serum PRL found very low prevalences of marked HPRL (>35 ng/ml) in ED patients (0.76% in a compilation of over 3200 patients) as well as of pituitary adenomas (0.4%). In addition, the association of HPRL with ED may have been coincidental in some of these cases, since 10% of the HPRLs diagnosed by the usual immunological assays are composed of macroprolactins, which are biologically inactive or little active variants of PRL. Their identification requires a PRL chromatography that is restricted to some specialized laboratories. There is presently no consensus with regards to the screening for HPRL in ED: systematic determination of serum PRL may be justified since HPRL is a serious but reversible disease, while there is presently no reliable clinical, psychometric or hormonal criteria (including serum testosterone level) allowing to restrict its determination to certain categories of the ED patients without risk of neglecting some HPRLs. In case of consistent HPRL, searching for a hypothalamic or pituitary tumor is mandatory. Dopamine-agonist therapy is the first choice treatment for the PRL-induced sexual dysfunctions. Additional sexual counselling may be necessary for certain patients. PMID- 14562141 TI - Epithelioid sarcoma of the penis--a rare differential diagnosis of Peyronie's disease. AB - We report on a case of penile epithelioid sarcoma in a 29-year-old man presenting with a dorsal penile plaque that primarily was misdiagnosed as Peyronie's disease. Although the initial clinical findings of these two different entities appear similar, the consequence for the patient is severe. The only way of differentiating these disorders are histological findings. The principal microscopic characteristics of epithelioid sarcoma are the distinctive nodular arrangement, central degeneration and necrosis of the tumor cells with epithelioid appearance and eosinophilia. Immunohistochemical data (cytokeratin, epithelial membrane antigen, vimentin, CD 34, desmin) confirm the diagnosis. We conclude that in cases with slightest doubts on the diagnosis of Peyronie's disease, especially in younger men suffering from a fast-growing penile induration, a bioptic clarification of the entity should be performed to exclude a high malignant disease that can be only treated as far as it is localized by radical surgery. PMID- 14562142 TI - Priapism associated with concurrent use of phosphodiesterase inhibitor drugs and intracavernous injection therapy. PMID- 14562143 TI - 'The limited practical value of color Doppler sonography in the differential diagnosis of men with erectile dysfunction' by Slob et al: a different point of view. PMID- 14562146 TI - Communicable diseases report, NSW, for May 2003. PMID- 14562147 TI - Wax models. PMID- 14562148 TI - Joseph Sampson Gamgee. PMID- 14562149 TI - History for undergraduates. PMID- 14562150 TI - "Let right be done": unprofessional conduct, reputations and injustices, and their effect on the life of the surgeon James Fitzjames Fraser West (1833-1883). AB - Unprofessional behaviour was still common when James Fitzjames Fraser West first qualified in 1854. The paper covers the ugly dispute that occurred in 1857, when West was appointed honorary surgeon at Queen's Hospital in Birmingham. West's reputation did not suffer any long-term harm, however, and he went to his grave a much-respected man. PMID- 14562151 TI - Percy Lane Oliver, OBE (1878-1944). PMID- 14562153 TI - Leonard Ralph Braithwaite (1878-1942). AB - Leonard Ralph Braithwaite lived and worked in Yorkshire all his life, with the exception of service overseas in World War I. He worked with Berkeley Moynihan early in his career, and went on to become a renowned surgeon and teacher in his own right. PMID- 14562152 TI - John Aitken (d. 1790)--grinder or scholar? AB - John Aitken attended the University of Edinburgh between 1763 and 1769 but did not graduate MD. He gained the membership (i.e. fellowship) of the Royal College of Surgeons of Edinburgh in 1770, and was for two sessions Senior President of the Royal Medical Society. Between 1771 and 1790 he published numerous books and pamphlets on surgery, medicine, midwifery, anatomy and physiology. As a surgeon at the Royal Infirmary, from 1779, he lectured on most subjects in the medical curriculum. John Struthers was particularly scathing of Aitken's scholarship, and this article attempts to restore Aitken's reputation as a scholar and probably one of the first of the extra-academical lecturers, who taught both anatomy and surgery in Edinburgh from 1779 until his death in 1790. PMID- 14562154 TI - Joseph Towne (1806-1879), master modeller of wax. AB - Joseph Towne (1806-1879) was a pioneer in the art of wax modelling of anatomical and dermatological material. He began his career at the age of 17, at Guy's Hospital, London, where he spent the whole of his professional life. He was a secretive, self-taught individual, and the finer points of his technique died with him. His brilliant works are on view at the Gordon Museum at Guy's, and are still used for teaching. PMID- 14562155 TI - Georges Clemenceau (1841-1929). PMID- 14562156 TI - Robert Hooke (1635-1703), in his own words. AB - The diaries and other writings of Robert Hooke (1635-1703), as well as those of his contemporaries, are drawn upon to sketch his social and scientific life. An account is presented of his involvement with the Royal Society from its earliest days, and of his relations with notable scientists. In exploring the similarity between combustion and respiration, he established that air is composed of different gases, and that it is not motion of the lungs but a supply of fresh air that is necessary for life. PMID- 14562157 TI - Greek professors of the Medical School of Constantinople during a period of reformation (1839-76). AB - A medical school was founded in Constantinople in 1827. Greek medics were involved with the new school right from its foundation, mainly because they had studied in Europe and knew other European and Asian languages. This paper reviews the lives of five of them: Stefanos Caratheodory; Constantinos Caratheodory; Sarantis Archigenis; Spyridon Mavrogenis; and Marco Pitsipio (Apostolidi Pasha), who was appointed a director of the Medical School. A few of the other Greek professors are also briefly discussed. PMID- 14562158 TI - What was the cause of Franklin Delano Roosevelt's paralytic illness? AB - In 1921, when he was 39 years of age, Franklin Delano Roosevelt contracted an illness characterized by: fever; protracted symmetric, ascending paralysis; facial paralysis; bladder and bowel dysfunction; numbness; and dysaesthesia. The symptoms gradually resolved except for paralysis of the lower extremities. The diagnosis at the onset of the illness and thereafter was paralytic poliomyelitis. Yet his age and many features of the illness are more consistent with a diagnosis of Guillain-Barre syndrome, an autoimmune polyneuritis. The likelihoods (posterior probabilities) of poliomyelitis and Guillain-Barre syndrome were investigated by Bayesian analysis. Posterior probabilities were calculated by multiplying the prior probability (disease incidence in Roosevelt's age group) by the symptom probability (likelihood of a symptom occurring in a disease). Six of eight posterior probabilities strongly favoured Guillain-Barre syndrome. PMID- 14562160 TI - Edward Jenner, MD FRS (1749-1823). PMID- 14562161 TI - Dr Heywood Smith's compound angulating gynaecological scissors. PMID- 14562162 TI - uPA, tPA and PAI-1 mRNA expression in periretinal membranes. AB - PURPOSE: Formation of periretinal membranes occurs in proliferative vitreoretinopathy (PVR) and proliferative diabetic retinopathy (PDR) and includes cell migration, proliferation, extracellular matrix formation and tissue contraction, processes in which plasminogen activation (PA) system is involved. METHODS: Twenty PVR, PDR or pucker membranes were examined to identify the cells with cell specific markers and to detect the expression of urokinase (uPA), tissue-type plasminogen activator (tPA) or plasminogen activator inhibitor-1 (PAI 1) by in situ hybridization and by immunohistochemistry. RESULTS: In PVR, uPA, tPA and PAI-1 were expressed by retinal pigment epithelial (RPE) cells, macrophages or retinal glial cells. In PDR, PA components were also expressed by endothelial cells. Semiquantitative analysis in in situ hybridization and immunohistochemistry results demonstrated no notable differences in uPA, tPA or PAI-1 expression between PDR and PVR membranes. CONCLUSIONS: We conclude that local proteolytic activation is involved in extracellular matrix production both in diabetic and non-diabetic membranes. PMID- 14562163 TI - Induction of cortical cataracts in cultured mouse lenses with H-89, an inhibitor of protein kinase A. AB - PURPOSE: To compare the effects of two serine-threonine protein kinase inhibitors in a mouse lens culture system previously designed to investigate cortical cataracts caused by L-buthionine sulfoximine (BSO), inhibitor of GSH biosynthesis. METHODS: Cataract development in HL-1 medium was evaluated visually or by measurement of lens Na+/K+ ratio through atomic absorption. Protein changes were evaluated by 32P-labeling, 2D-gel electrophoresis, phosphorimaging and mass spectrometry. Results. H-7 (50 microM), inhibitor of protein kinase A (PKA) and protein kinase C (PKC), did not cause cataracts, but inhibited BSO cataract development. By contrast, 25 microM H-89, selective inhibitor of PKA, caused large annular cortical cataracts and 100-fold elevation of Na+/K+ within 30 hr in day 10 lenses, in either the presence or absence of BSO. H-89 cataracts were also seen in day 12 and day 21 lenses. 32P-labeling of day 12 lenses pretreated with H 89 displayed more than 80% decrease in phosphorylation of alphaA crystallin, a known substrate of PKA, in the insoluble protein fraction. 2D-gel electrophoresis of day 12 H-89 cataract lens fractions revealed limited degradation of alpha and beta crystallins, degradation of cytoskeletal proteins, and elevated lens Ca2+ (>4 nmol/mg wet wt.), suggesting Ca2+-activated proteolysis. Conclusions. High Na+/K+ cataracts are induced by H-89, selective inhibitor of PKA, but not by H-7, an inhibitor of both PKA and PKC that impeded BSO-induced Na+/K+ elevation and cataract. These results suggest contrasting effects of PKA and PKC on lens cation transport and cortical cataract development. PMID- 14562164 TI - Expression and functional properties of unique inward rectifier K+ channel Kir7.1 in the porcine iris and retinal pigment epithelium. AB - PURPOSE: To investigate the membrane functional properties of porcine iris pigment epithelial cells (IPE), and compare the characters of inward rectifier potassium (Kir) channel in the IPE with those in the retinal pigment epithelial cells (RPE). METHODS: IPE and RPE were acutely dissociated from porcine eyes. Functional properties of Kir channels were characterized using whole cell patch clamp recording techniques. Expression of Kir7.1 mRNA in both cells was detected by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Whole cell current in the IPE exhibited a mild inward K+ rectification, and showed little dependence on [K+]o. Unusual high (7.04 +/- 1.7) Rb+ to K+ inward conductance ratio indicated that Kir7.1 subunit was expressed in the IPE as the same as RPE cells. Also, Kir7.1 mRNA was detected in both porcine IPE and RPE by RT-PCR. However, functional expression of Kir conductance in IPE cells (21.7 S/F) was much smaller than that in RPE cells (205.6 S/F). CONCLUSIONS: The Kir7.1 subunit was predominantly expressed in the acutely dissociated porcine IPE and its functional properties are similar to those in the RPE. However, the current density seems too small to fulfill the task of the Kir function of RPE. PMID- 14562165 TI - A comparison of invasive and cytotoxic Pseudomonas aeruginosa strain-induced corneal disease responses to therapeutics. AB - PURPOSE: During corneal infection, cytotoxic Pseudomonas aeruginosa strains remain mostly extracellular, while invasive strains can enter corneal cells and replicate within them. We tested the hypothesis that ofloxacin, which easily penetrates host cell membranes, would be more effective than the less cell permeable antibiotic tobramycin, for treatment of corneal infection by an invasive P. aeruginosa strain. METHODS: A murine model of P. aeruginosa keratitis was used to compare the response to ofloxacin, tobramycin, prednisolone acetate, and non-preserved saline treatment, as well as combination antibiotic corticosteroid therapy for infection caused by a cytotoxic strain (6206) and an invasive strain (PAO1). Treatment involved hourly eye drop administration for 12 hours. RESULTS: As expected, tobramycin was less effective at eradicating viable bacteria from corneas infected with the invasive strain. Despite rapid sterilization of corneas in other antibiotic treated groups, disease progression occurred during the 12 hour treatment period. Both antibiotics hastened disease resolution over the next 7 days for infections caused by either strain. Corticosteroid use during the 12 hour treatment period was of little added benefit. CONCLUSIONS: Differences between invasive and cytotoxic strain infections in their early response to the different therapeutic regimens did not translate to notable differences after 7 days, but the effects of antibiotics in halting disease progression were delayed for both strain types. These results suggest that successful management might be improved by addressing factors contributing to disease progression during sterilization of the cornea by antibiotics. PMID- 14562166 TI - Heme oxygenase in the retina in diabetes. AB - INTRODUCTION: Heme oxygenase (HO) isoforms, HO-1, and HO-2, are responsible for heme breakdown to iron and carbon monoxide (CO). HO may respond to oxidative stress and may modulate the expression of vasoactive factors like nitric oxide (NO). Since diabetes induced oxidative stress may change HO, the present study examined whether diabetes is associated with HO alterations, its relationship with NO, endothelin-1(ET-1) and the functional significance. MATERIALS AND METHODS: Male SD rats with Streptozotocin induced diabetes were investigated after six-weeks. Poorly controlled diabetic animals were randomized to one of three treatment groups (n = 6 each group); a) untreated, b) HO-1 inhibitor SnPP IX (50 micromol/kgIP/day), c) NO donor molsidomine (120 mg/L PO/day) and were compared with age and sex matched non diabetic control animals with or without SnPP-IX treatment. Color Doppler ultrasound analysis was used to determine retinal resistivity index (RI). mRNA for HO-1, HO-2, ET-1, eNOS and iNOS were analyzed with competitive RT-PCR. HO distribution in the retina was investigated by immunocytochemistry. RESULTS: Diabetic animals expressed lower body weight, higher blood glucose and increased glycated hemoglobin levels. HO-1 and HO-2 immuno-reactivity were identified in the retina. Diabetes induced increased RI was associated with up-regulation of both ET-1 and HO-1 mRNA expression but not eNOS or iNOS mRNA. Both SnPP-IX and molsidomine treatments prevented a diabetes increase of RI, in spite of increased ET-1 expression and were associated with increased iNOS mRNA. CONCLUSIONS: The present data suggests that the HO system is up-regulated in short term diabetes leading to HO and NO interactions which may modulate vascular function in the retina. PMID- 14562167 TI - Ibuprofen improves oxygen-induced retinopathy in a mouse model. AB - PURPOSE: Retinopathy of prematurity is a developmental vascular anomaly occurring in the incompletely vascularized retina of the premature infant. Ibuprofen is a nonsteroidal anti-inflammatory agent similar to indomethacin, but with less pronounced side-effects. The goal of the study was to test the hypothesis that ibuprofen would improve oxygen-induced retinopathy in a mouse model. METHODS: C57BL6 mice pups were exposed to 75% oxygen from postnatal day 7 through postnatal day 12. Ibuprofen was administered along with oxygen exposure as a single subcutaneous dose of 40 mg/kg/day for 5 days. Animals were sacrificed on postnatal day 17 through postnatal day 20. The severity of retinopathy was assessed by a retinopathy scoring system of fluorescein-conjugated dextran perfused retinal flat mounts and by quantitation of extra-retinal nuclei by use of periodic acid-Schiff-stained retinal sections. RESULTS: Animals that received ibuprofen during hyperoxia exposure had a significantly lower median (25th, 75th quartile) retinopathy score of 6 (5, 7.5) compared with animals that received oxygen only, with a score of 12 (10.5, 12.5), with p < 0.005. Animals given ibuprofen during hyperoxia exposure had a significantly lower extra-retinal nuclei count per section (14.2 +/- 3.6) compared with animals that were only exposed to oxygen (26.8 +/- 5.8), with p < 0.005. Ibuprofen did not affect the growth of the animals. CONCLUSION: Ibuprofen improves oxygen-induced retinopathy when administered concurrently with the injury phase without affecting the normal retinal development of the animals. PMID- 14562168 TI - Senescent effects on binocular summation for contrast sensitivity and spatial interval acuity. AB - PURPOSE: Age effects on binocular summation, a cortically mediated visual function, were compared for resolution acuity, contrast sensitivity (CS), and spatial interval (SI) hyperacuity. METHOD: The binocular and monocular thresholds of healthy, optimally-corrected healthy young (mean age 21.3 years) and old (mean age 69.9 years) observers were determined for acuity, SI discrimination, and CS at 1.0, 2.0, 4.0, 8.0 and 18.0 c/deg. RESULTS: No age effects were observed on monocular or binocular SI discrimination. The binocular summation ratios (BSRs) did not exceed the expected probability gain for resolution acuity or SI discrimination in either age group. Older observers showed a binocular inhibition effect on the SI task. On the CS task, the BSRs of the young significantly exceeded those of the old only at 18.0 c/deg. CONCLUSIONS: 1.) Binocular summation of high spatial frequency contrast information, which may be less robust in the senescent visual system, did not appear to be related to interocular CS differences, 2.) SI discrimination of high contrast, well separated targets appears to be unaffected by aging, and 3.) Binocular neural summation on SI discrimination may be more likely to be seen with targets that are narrowly separated or low in contrast. PMID- 14562169 TI - Levels of mucin gene expression in normal human conjunctival epithelium in vivo. AB - PURPOSE: Conjunctival impression cytology (CIC) samples were used to determine the mean and normal range of mRNA levels of human MUC1, MUC2, MUC4, MUC5AC, and MUC7 mucin genes. METHODS: Real time PCR was performed to determine normal mRNA levels in CIC samples of 24 male and 19 female healthy donors. Correlation coefficients between gene expression levels were obtained. RESULTS: All five mucin genes were expressed in the CIC samples. MUC1 and MUC4 were present at the highest level and MUC2 was at the lowest. There were no gender differences. Significant positive correlations existed between MUC2 and MUC4 and between MUC2 and MUC7 levels. CONCLUSIONS: Normal levels and ranges of mRNAs for MUC1, MUC2, MUC4, MUC5AC and MUC7 conjunctival mucin genes have been established for the first time. These data may serve as the normal threshold values for future comparisons in different experimental and pathological conditions involving the ocular surface. PMID- 14562170 TI - Prolonged exposure to high glucose impaired cellular behavior of normal human corneal epithelial cells. AB - PURPOSE: To determine the effect of prolonged exposure to high glucose on cellular behavior of normal human corneal epithelial cells (HCEC). METHODS: HCEC were cultured in medium under normal or high glucose conditions for 14 days. Proliferation was evaluated by direct cell counting and [(3)H]thymidine incorporation. Cell cycle analysis was performed using flow cytometry. The ability of HCEC to attach to type I collagen was evaluated using a short-term colorimetric adhesion assay. The effect of high glucose on the expression of integrin alpha(3)beta(1) was also evaluated using flow cytometry. RESULTS: Cell number and [(3)H]thymidine incorporation under high glucose conditions decreased compared with those under normal glucose conditions. The cells exposed to high glucose were G(0)/G(1) than untreated cells. The adhesion ability of HCEC under high glucose conditions decreased compared to normal glucose conditions. Expression of integrin alpha( 3)beta(1) was down-regulated under high glucose conditions. CONCLUSIONS: High glucose had deleterious effects on cellular behavior of HCEC, which might cause delayed corneal epithelial wound healing in diabetic keratopathy. PMID- 14562171 TI - Characterization of serotonergic receptors in rabbit, porcine and human conjunctivae. AB - PURPOSE: To characterize the serotonin (5-HT) receptors linked to the modulation of adenylyl cyclase activity in rabbit, porcine and human conjunctivae. METHODS: Serotonin receptor-subtype expression was examined using reverse transcription polymerase chain reaction (RT-PCR) and receptor subtype-specific polyclonal antibodies for the immunofluorescent labeling of conjunctival cryosections. In addition, measurements of the effects of serotonergics on the short-circuit current (I(sc)) across rabbit and porcine conjunctivae were contrasted. RESULTS: RT-PCR assays indicated the expression of 5-HT(1B ) and 5-HT(1D) receptors, subtypes negatively coupled to adenylyl cyclase, in the rabbit conjunctiva. This approach also suggested the co-expression of 5-HT(1B), 5-HT(1D), 5-HT(1F), 5 HT(4) and 5-HT(7) mRNA's in the porcine conjunctiva, and 5-HT( 1D), 5-HT(1F) and 5-HT(7) in the human conjunctiva. Since the 5-HT(4) and 5-HT(7) receptors are positively linked to adenylyl cyclase, these results implied that the porcine and human tissues exhibited subtypes both positively and negatively linked to the enzyme. However, immunohistochemical observations, using currently available antibodies solely localized the 5-HT(7) moiety in the porcine and human epithelia, suggested that the 1B/1D forms may be minor elements. Consistent with this prospect, 5-HT was a stimulant of the transepithelial I(sc) across the porcine conjunctiva, an opposite response from earlier findings that demonstrated inhibitory effects by 5-HT on the rabbit I(sc), which are now explained by the localization of the 1B/1D receptors in the rabbit stratified epithelium. CONCLUSIONS: The 5-HT receptors expressed by mammalian conjunctivae are not identical. In terms of 5-HT receptor expression, the porcine tissue may be a more appropriate model for human, than is the rabbit, in that 5-HT may serve as a secretagogue in the human epithelium. PMID- 14562172 TI - Myogenic response in isolated porcine retinal arterioles. AB - PURPOSE: To study the myogenic response in large and small porcine retinal arterioles and the effects of blockade of L-type voltage gated calcium channels on the myogenic response. METHODS: Eleven large (outer diameter 137.7 +/- 4.5 micro m) and fourteen small (77.3 +/- 3.7 micro m) isolated porcine retinal arterioles were studied for myogenic response while mounted between two double barrelled pipette systems. The intraluminal pressure was varied from 0 to 140 cm H( 2)O (0-103 mmHg) and the diameter changes of the arterioles were registered. The pressure-diameter relation was determined for arterioles in calcium containing and calcium free solutions and during inhibition of the L-type voltage gated calcium channels with nifedipine. RESULTS: Both large and small retinal arterioles showed a decrease in diameter as the intraluminal pressure was increased, but no significant difference was found between large and small arterioles (p = 0.39). Blockade of voltage gated calcium channels significantly abolished the myogenic response (p = 0.010 and p = 0.005, large and small arterioles respectively). CONCLUSIONS: The results indicate that myogenic responses are present throughout the retinal arteriolar system and that L-type voltage gated calcium channels are involved in transforming the stretch of retinal vascular smooth muscle cells induced by an increase in intraluminal pressure into a contraction. PMID- 14562173 TI - Exclusion of the human collagen type XVII (COL17A1) gene as the cause of Thiel Behnke corneal dystrophy (CDB2) on chromosome 10q23-q25. AB - PURPOSE: Determination of the gene causing Thiel-Behnke Corneal Dystrophy (CDB2) would have important clinical implications. Previous studies in our laboratory have suggested that the COL17A1 gene may be the cause of Thiel-Behnke Corneal Dystrophy (CDB2) on Chromosome 10q23-q25. METHODS: We evaluated a five-generation family with CDB2 mapped to chromosome 10. Many of these family members were diagnosed by slit-lamp microscopy. In addition, genomic DNA was isolated and purified from peripheral blood samples. The COL17A1 gene was screened for possible disease causing mutations by PCR and DNA sequencing analysis. RESULTS: No disease-causing mutations were found in any of the 56 exons of the COL17A1 gene or in any of the flanking intron/exon junctions. CONCLUSIONS: Mutations in the coding sequence of the human collagen XVII (COL17A1) gene are not the cause of CDB2. PMID- 14562174 TI - The effect of protein-coated contact lenses on the adhesion and viability of gram negative bacteria. AB - PURPOSE: Gram negative bacterial adhesion to contact lenses can cause adverse responses. During contact lens wear, components of the tear film adsorb to the contact lens. This study aimed to investigate the effect of this conditioning film on the viability of bacteria. METHODS: Bacteria adhered to contact lenses which were either unworn, worn for daily-, extended- or overnight-wear or coated with lactoferrin or lysozyme. Numbers of viable and total cells were estimated. RESULTS: The number of viable attached cells was found to be significantly lower than the total number of cells on worn (50% for strain Paer1 on daily-wear lenses) or lactoferrin-coated lenses (56% for strain Paer1). Lysozyme-coated lenses no statistically significant effect on adhesion. DISCUSSION: The conditioning film gained through wear may not inhibit bacterial adhesion, but may act adversely upon those bacteria that succeed in attaching. PMID- 14562175 TI - Macrophages and microcirculation in regressed and partially regressed irradiated choroidal and ciliary body melanomas. AB - PURPOSE: To investigate how tumour-infiltrating macrophages and microcirculation attributes of uveal melanomas regressed after brachytherapy and whether primarily enucleated melanomas differ. METHODS: A case-control analysis of 34 matched pairs of irradiated and nonirradiated choroidal and ciliary body melanomas with main outcome variables being area of necrosis, extravascular matrix loops and networks, tumour-infiltrating macrophages in nonnecrotic areas identified with mAb PG-M1 to the CD68 epitope, and microvascular density (MVD) determined by mAb QBEND/10 to the CD34 epitope. RESULTS: Comparison of primarily enucleated eyes to eyes with irradiated, secondarily enucleated melanomas revealed significantly more necrosis (median difference, +9%, P = 0.0012) and lower MVD (median difference, -10 counts/0.313 mm(2), P = 0.011) in the latter. In eyes managed with brachytherapy, loops and networks tended to be less frequent (P = 0.077). Number and type of macrophages were similarly distributed, being moderate to high in about 95% (P = 0.67) of the matched pairs, and intermediate to dendritic in 79% (P = 0.90). In the irradiated eyes, presence of epithelioid cells and the number and type of macrophages showed no association with microcirculation attributes, whereas in the primarily enucleated tumours, high number of macrophages was associated with high MVD (P < 0.001). CONCLUSIONS: This study suggests that regression after brachytherapy reduces MVD. The difference cannot be attributed to different numbers of tumour-infiltrating macrophages and different cell type in nonnecrotic areas of the tumour. PMID- 14562176 TI - Fluorescein transport through the blood-aqueous and blood-retinal barriers in diabetic macular edema. AB - PURPOSE: To investigate the integrity of the blood-aqueous barrier (BAB) and the blood-retinal barrier (BRB) in diabetic patients with clinically significant macular edema (CSME). METHODS: The rate constant (Kd(F)) of the BAB and the permeability of the blood-retinal barrier for non-protein bound fluorescein were determined simultaneously by fluorometry. Results were analysed pairwise in diabetic patients (n = 25) with CSME in one eye and without CSME in the other eye. RESULTS: Kd(F) for the eyes with CSME was significantly increased compared to eyes without CSME (444. 10(-6) min( -1) and 387. 10(-6) min(-1) respectively, p = 0.01). The passive permeability of the BRB was also significantly increased in CSME (5.7 nm/sec and 3.5 nm/sec respectively, p = 0.009, n = 19). CONCLUSIONS: Both the rate constant of the BAB and the BRB permeability were significantly increased in CSME indicating that eye-specific factors are common for both barriers in diabetic patients with CSME. PMID- 14562177 TI - In vitro effects of preserved or preservative-free antiglaucoma medications on human complement system. AB - PURPOSE: Antiglaucoma drugs have been associated with conjunctival and trabecular inflammatory cell infiltrates. However, the underlying mechanisms are still poorly understood. The aim of this study was to assess the effects of antiglaucoma medications on the complement system, an early mediator of the inflammatory response. METHODS: Human serum was first treated with a classical or alternative pathway activator (aggregated human IgG or zymosan, respectively) in the presence or the absence of preservative-free or benzalkonium (BAK)-preserved antiglaucoma drugs. CH50 assay was then performed to assess the functional activity of residual complement in treated serum. RESULTS: In the absence of complement activator, the antiglaucoma drugs tested in this study were all devoid of intrinsic complement-activating potency. Preserved and preservative-free carteolol as well as preserved latanoprost did not worsen or prevent complement activation induced by zymosan or aggregated IgG. Unexpectedly, both preserved and unpreserved timolol and betaxolol significantly counteracted the effects of complement activators. Timolol prevented activation triggered by both IgG and zymosan to the same extent (24% to 29%), despite the presence of BAK in the preserved formulation. Betaxolol was twice as effective at preventing the effect of IgG (34% to 37%) than that of zymosan (14%), regardless of the presence of BAK. However, BAK itself strongly aggravated complement activation by both activators. CONCLUSIONS: Carteolol, timolol, betaxolol and latanoprost did not activate complement system. On the contrary, the beta-blockers timolol and betaxolol exerted an anti-inflammatory effect by preventing complement activation. The deleterious effect of benzalkonium seems to have been neutralized within the preserved eyedrops through a mechanism that remains to be elucidated. Our study suggests that inflammatory signs in glaucoma patients should not be attributed to complement activation by antiglaucoma drugs. PMID- 14562178 TI - Phosphorylation of MAP kinase in corneal epithelial cells during wound healing. AB - PURPOSE: To investigate the role of mitogen-activated protein kinase (MAPK), such as p44/42 MAPK, p38 MAPK and stress-activated protein kinase (SAPK), in corneal epithelial cells during the wound healing process. METHODS: A single non penetrating incision was produced on rat cornea. Then the corneal wound healing process was observed with an immunocytochemical technique using specific antibodies reacting only with phosphorylated p44/42 MAPK, p38 MAPK or SAPK. Cell lysates of corneal epithelial cells in rabbits stimulated with keratinocyte growth factor (KGF) and hepatocyte growth factor (HGF) were processed for Western blot using antibodies to phosphorylated p44/42 MAPK. RESULTS: Maximum activation of p44/42 MAPK was observed in wing and basal cells at wounded regions in rat cornea at 1 hour after the incision. Activation of p44/42 MAPK was still detected in all basal and wing cells at wounded regions at up to 24 hours when the incisions were completely closed, and then receded to normal intensity after 7 days. Neither p38 MAPK nor SAPK were activated during the wound healing process. Western blot analysis of cultured corneal epithelial cells in rabbits showed phosphorylation of p44/42 MAPK after 30 minutes in response to KGF and HGF, whereas non-activated p44/42 MAPK was ordinarily detected even at the absence of KGF or HGF. CONCLUSIONS: These results demonstrate that p44/42 MAPK is activated during the corneal wound healing process and suggest that KGF and HGF play an important role in initiation of cell migration and proliferation in the initial wound healing process by activating p44/42 MAPK. PMID- 14562179 TI - Revised formulas for summarizing retinal vessel diameters. AB - Background/Purpose. Recent findings suggest that an objective assessment of retinal vessel caliber from fundus photographs provide information about the association of microvascular characteristics with macrovascular disease. Current methods used to quantify retinal vessel caliber, introduced by Parr(1,2) and Hubbard,(3) are not independent of scale and are affected by the number of vessels. To improve upon these methods we introduce revised formulas for quantifying vessel caliber. Methods. Revised formulas were estimated using retinal vessel measurements from 44 young adults free of hypertension and diabetes. Comparisons between the two methods were done using digitized photographs from 4926 participants at the baseline examination of the Beaver Dam Eye Study (BDES), an ongoing population-based cohort study initiated in 1987. Individual arterioles and venules were measured using semi-automated computer software from which summary measures were calculated. Results. Correlation coefficients between the Parr-Hubbard and revised formulas were high (Pearson correlation coefficients ranging from 0.94 to 0.98). Both arteriolar and venular caliber significantly increased with an increasing number of vessels measured using the Parr-Hubbard formulas (p < 0.001), which in turn affected the relationship to mean arterial blood pressure. To the contrary, the revised formulas were not affected by the number of measured vessels (p > 0.50). Conclusions. We describe revised formulas for summarizing retinal vessel diameters measured from fundus photographs to be used in future studies and analyses. The revised formulas correlate highly with the previously used Parr Hubbard formulas, but offer the advantages of being more robust against variability in the number of vessels observed, being independent of image scale, and being easier to implement. PMID- 14562180 TI - HLA class II antigen expression in conjunctival precancerous lesions and squamous cell carcinomas. AB - PURPOSE: The expression of human leukocyte antigen (HLA) class II molecules on the cell surface is necessary for the presentation of peptide antigens to helper CD4 T lymphocytes of the immune system. We studied the immunoexpression of HLA class II antigen in conjunctival precursor lesions and conjunctival squamous cell carcinomas. METHODS: HLA class II antigen expression was analyzed in 8 conjunctival dysplasias, 6 carcinomas in situ and in 7 conjunctival squamous cell carcinomas, by immunoperoxidase staining with monoclonal antibody to HLA class II antigen on the archival clinical samples. Immunoanalysis was done by a semi quantitative method based on the intensity of staining and the percentage of stained cells. RESULTS: HLA class II antigen immunoexpression was heterogeneous in 8 conjunctival dysplasias and in 6 carcinoma in situ and negative in 7 conjunctival squamous cell carcinomas. CONCLUSIONS: Human leukocyte class II antigen immunoexpression is decreased in conjunctival precancerous and squamous cell carcinomas. PMID- 14562181 TI - Microinjection of actin antibodies impaired gap junctional intercellular communication in lens epithelial cells in vitro. AB - PURPOSE: The aim of this study was to check the importance of cytoskeletal actin for gap junction mediated intercellular communication (GJIC) in cultured lens epithelial cells (LEC). METHODS: Bovine LEC were cultured until confluency on cover-slides of a collocate-system. In order to study the cytoskeletal influence on cell communication microcinjection of gap junction permeable neurobiotin into a single cell was preceded by microinjection of actin antibodies. Confocal laser scanning microscopy of specimens treated with actin antibodies and/or subsequent phalloidin labelling, and electron microscopy, were applied to check for cytoskeleton cell membrane links. Specificity of actin antibodies was proved by immoblotting techniques. RESULTS: Immunohistochemistry and phalloidin-rhodamine staining displayed bundles of actin-filaments extending through the entire LEC. Quantitative analysis of GJIC showed intensive dye-spreading of neurobiotin between adjacent LEC. Injection of actin antibodies thirty minutes prior to microinjection of neurobiotin significantly reduced GJIC. Microinjection of irrelevant antibodies had no effect on GJIC. CONCLUSION: Integrity of the actin cytoskeleton is fundamental for unimpaired GJIC in LEC. PMID- 14562182 TI - Distribution of thrombospondin-4 in the bovine eye. AB - PURPOSE: The purpose of this study was to analyze the distribution of thrombospondin-4 (TSP-4) in the bovine eye. METHODS: Anterior and posterior segments of the bovine eyes were sectioned and stained by the indirect immunofluorescence method with an anti-TSP-4 antibody. The tissues were analyzed by reverse-transcription-polymerase chain reaction (RT-PCR) to determine where the TSP-4 mRNA is produced. RESULTS: Immunohistochemical staining for TSP-4 indicated the presence of TSP-4 in the cornea (epithelium, basement membrane, and keratocytes), conjunctiva (epithelium and stroma), aqueous ducts, sclera, iris (stroma), ciliary processes and muscle, trabecular meshwork, Bruch's membrane, retina, lamina cribrosa, and optic nerve, and in all blood vessel walls. TSP-4 mRNA was expressed by the cells in all structures. CONCLUSIONS: TSP-4 is widely distributed in the bovine eye where it may play a role in the functions of basement membranes in various tissues. It is abundant in the trabecular and uveo scleral pathways and may play a role in the regulation of aqueous outflow resistance. PMID- 14562183 TI - A-scan quantitative echography of senile cataracts and correlation with phacoemulsification parameters. AB - PURPOSE: To study the echographic appearance of various types of senile cataracts using a-scan quantitative echography and correlate these findings with phacoemulsification parameters indicating surgical lens hardness. METHODS: The study involved 71 eyes with senile cataracts and 20 additional eyes with normal human lenses. Cataract and clear lenses were studied using A-scan quantitative echography and with a tissue sensitivity gain setting of 74 db. We assessed the lens internal reflectivity in the A-scan echograms within a range of 0% to 100%, and calculated the mean of all spikes. All eyes with cataract underwent clear corneal phacoemulsification. At the end of the operation we recorded the phacoemulsification time and mean percent power. We determined the correlations of echographic and phacoemulsification data. RESULTS: Quantitative echography in the 71 studied eyes with cataract revealed acoustic reflections having various configurations according to the biomicroscopic type of cataract, i.e. cortical, nuclear, corticonuclear, brunescent, white intumescent. The mean of all echographic spikes within the cataract lenses varied from 3% (in some brunescent very sclerotic lenses) to 53%. Echographic reflections were not observed in the A scan echograms of 20 additional eyes with normal human lenses. The mean of internal lens spikes correlated negatively with phacoemulsification time (p < 0.001; r = -0.703) and mean percent phacoemulsification power (p < 0.001; r = 0.57). CONCLUSIONS: The results indicate that A-scan quantitative echography may be used as an adjunctive tool in the preoperative evaluation of cataracts, particularly in cases with compromized visualization of the lens (small pupil, corneal opacities). PMID- 14562184 TI - Evolving neurovascular relationships in the RCS rat with age. AB - PURPOSE: To examine the course of development of vascular disorders in the Royal College of Surgeons (RCS) rat and how these may lead to retinal ganglion cell loss. METHODS: Whole-mount retinae from RCS rats were first stained for neurofilament protein and then for NADPH-diaphorase staining. A separate group of RCS rats was injected with Type II Peroxidase and the retinae were subsequently processed for peroxidase histochemistry. RESULTS: The first changes in the deep vascular plexus occur as the photoreceptor layer is lost and it comes into close proximity to the retinal pigment epithelial (RPE) cell layer. RPE cells migrate onto retinal vessels, and at such locations vascular complex develop. These are first found ventral to the optic nerve head and then gradually progress over most of the retina. The inner retinal vessels that supply the complexes cross the optic nerve fiber layer and appear to be under tension. They ligate axons, which leads to retinal ganglion cell loss. CONCLUSIONS: These observations show vascular changes can have secondary repercussions for neurons distant from the primary lesion. PMID- 14562185 TI - Quality requirements in clinical studies: a necessary burden? AB - Drug development has continued to progress over the past decades and the processes involved have become more complex. The basis for these changes include advances in substance analysis and production, targeted drug development and defined guidelines for the conduct of clinical trials. The aim of standardised quality requirements is to generate scientifically sound data in clinical trials while observing strict ethical rules. The conduct of clinical trials is governed by internationally established Good Clinical Practice (GCP) guidelines. For the protection of human beings directly involved, framework conditions must be established as guidelines to accompany the research process. At the same time, these regulatory requirements enable a quality criteria standard to be defined. However, those regulations form a major challenge for clinical study centres, since they affect current clinical practice considerably. The objective of this paper is, on the one hand, to clarify the historical background relating to the development of clinical trials and the corresponding legal guidelines, on the other hand, to discuss consequences for clinical practice. PMID- 14562186 TI - Hand function assessment in patients receiving haemodialysis. AB - OBJECTIVE: To assess the usefulness of Duruoz's Hand Index (DHI) in patients undergoing haemodialysis. METHODS: Patients receiving haemodialysis for more than 2 months were recruited randomly. Demographic, clinical and functional characteristics of patients were evaluated. Functional assessment was performed with DHI, Hand Functional Index (HFI), Health Assessment Questionnaire (HAQ), Purdue Pegboard, grip strength and 3 kinds of pinch strengths. DHI was correlated (Spearman's) with the other functional parameters in assessing the convergent validity and with non-functional parameters in assessing the divergent validity. RESULTS: Sixty patients with a mean age of 50.05 were recruited. The average duration of haemodialysis was 55.02 months. DHI is significantly correlated with HAQ, HFI, Purdue Pegboard scores, grip strength and 3 types of pinch strengths while no significant correlation was found with non-functional parameters. CONCLUSIONS: DHI is a practical scale which is efficient in assessing accurately the functional disability of the hand in patients receiving haemodialysis. PMID- 14562187 TI - Elevated troponin levels in absence of coronary artery disease after supraventricular tachycardia. AB - Generally speaking elevated troponin levels are consistent with the diagnosis of acute coronary syndrome and haemodynamically relevant coronary artery stenosis. However, they may also point to minor myocardial injury in other circumstances. Four patients with elevated troponin levels after supraventricular tachycardia without evidence of coronary artery disease and very low risk scores for acute coronary syndrome are described and discussed. PMID- 14562188 TI - Determination of RHD zygosity using real-time quantitative PCR. AB - At present RHD incompatibility is still an obstetric problem despite prophylactic treatment. A very welcome recent technical advance has now made it possible to determine the foetal RHD status in a non-invasive risk-free manner using cell free foetal DNA in maternal plasma. In some cases, however, where there is a high risk that the foetus may be affected by HDN (haemolytic disease of the newborn), it may be of interest to determine whether the father is hetero- or homozygous for the RHD gene, since in the former instance there is only a 50% chance that the pregnancy is affected. It has recently been shown that quantitative PCR assays, in particular real-time Taqman PCR, can be used to determine the RHD gene dosage, and also to determine foetal aneuploidies. We demonstrate that the same real-time Taqman PCR assay we had previously developed for non-invasive analysis of the foetal RHD gene and the foetal Y chromosome from maternal plasma can be used to determine the paternal RHD genotype. PMID- 14562189 TI - Status epilepticus in a case with wilson's disease during D-pencillamine treatment. PMID- 14562190 TI - Idiopathic pulmonary fibrosis--a disorder of alveolar wound repair? AB - Idiopathic pulmonary fibrosis (IPF) is a chronic and usually progressive lung disorder of unknown aetiology. Conventional management of patients with IPF has been primarily based on the concept that suppressing inflammation would prevent progression to fibrosis. Although the pathogenesis is incompletely understood, it is here suggested that IPF is a disease of abnormal wound repair and remodelling in the lung rather than an inflammatory disease. Therefore, treatment strategies are no longer aimed at reducing inflammation, but rather at preventing or inhibiting the fibroproliferative responses and enhancing efficient alveolar epithelial repair. So far, no cell-specific drugs for these purposes are clinically available. However, novel promising molecules or drugs are being studied in experimental models or ongoing clinical trials in patients with IPF. Evolving hypotheses on the pathogenesis of IPF are reviewed, focusing on possible implications for future therapies. A better understanding of the sequence of the pathogenic mechanisms that control the fibrotic response will hopefully lead to efficient therapies and finally a favourable outcome in patients with this disease. PMID- 14562191 TI - A new animal model of cerebral venous infarction: ligation of the posterior part of the superior sagittal sinus in the cat. AB - QUESTIONS UNDER STUDY: Dural sinus occlusion is an infrequent but potentially devastating cause of stroke. The pathophysiological course of events underlying it is, as yet incompletely understood. METHODS: In a cat model, regional cerebral blood flow (CBF) was measured during control and 2, and 24 hours after superior sagittal sinus occlusion. Around 48 hours after superior sagittal sinus occlusion, experimental settings were terminated by perfusion fixation with 4% paraformaldehyde solution, and haematoxylin and eosin histology. RESULTS: CBF was significantly reduced over the time-period of measurement (p < 0.05) covering about 45% of the brain in planes that were affected by occlusion. Histologically, in all cases signs of subacute venous infarction could be demonstrated. CONCLUSIONS: Based on the newly-developed model of microsurgical ligation of the superior sagittal sinus in cats, we present for the first time an animal model for cerebral venous infarction that leads to a histologically proven subacute venous infarction with a good reproducibility. The further advantage of this model is the fact that it mimics the clinical situation as far as possible by its inter- and intra-individual variance of extension of the venous infarction and by the slow reduction of CBF over 24 hours. Sequential PET imaging is a favourable, non-invasive method to gain further insight into the pathophysiological characteristics of experimental cerebral venous infarction. Therefore, the new developed cat-model as demonstrated in this study will be of great value for further and more detailed investigations of cerebral-venous infarctions, and for the experimental evaluation of therapeutic strategies. PMID- 14562192 TI - Long-term glucose insulin potassium infusion improves systolic and diastolic function in patients with chronic ischemic cardiomyopathy. AB - We assessed the effects of glucose-insulin-potassium (GIK) using echocardiography, right ventricular catheterisation and myocardial scintigraphy with 99mTc sestamibi in stable patients with ischaemic cardiac dysfunction. METHODS: Thirty male patients with stable coronary disease (SCD) and an ejection fraction (EF) <40% were studied for systolic and diastolic function. Glucose 30%, 300 insulin units and KCl 6 g/l were infused at 1 ml/kg per hour for 24 hours. Haemodynamic, echocardiographic and myocardial scintigraphy measurements were recorded at rest on completion of the GIK infusion. RESULTS: A significant increase in EF was observed (32.1 +/- 7.8% and 43.3 +/- 11.6%, p <0.01). A significant prolongation was seen in the diastolic filling periods (365 +/- 52 msec and 428 +/- 70 msec, p <0.05). A significant decrease in pulmonary capillary wedge pressure was measured (22.2 +/- 5.3 and 17.1 +/- 4.3, p <0.01) and a significant decrease in stress score (SS) was observed (13 +/- 7 and 11 +/- 5, p >0.05). CONCLUSION: Our present work suggests that GIK infusion improves systolic and diastolic function in patients with SCD and an ejection fraction <40%. Further studies are needed to determine whether long-term GIK infusion could be useful for therapeutic strategies in patients with chronic ischaemic coronary diseases. PMID- 14562193 TI - Remodelling in young sheep: a histological study of experimentally produced defects in TMJ. AB - QUESTIONS UNDER STUDY: In this study, experimentally induced surgical defects of the mandibular condyle were evaluated histologically 3, 6 and 9 months postoperatively to investigate the capacity for remodelling which is one of the most important features of the temporomandibular joint. METHODS: In three healthy sheep standardised surgical defects were produced on each right condylar head. The left temporomandibular joints served as controls. No treatment was performed on the surgical defects and function was not restricted postoperatively. At the end of the experimental period the sheep were sacrificed and a histological evaluation was carried out. RESULTS: Histologically, no disturbance was observed during the healing process in bone and cartilage. Although function was not limited throughout the study, it was observed that the experimental surgical defects had healed completely. CONCLUSIONS: This result points to the condyle's high capacity for repair and remodelling. PMID- 14562194 TI - [Validation of a German Version of the Fatigue Impact Scale FIS-D]. AB - OBJECTIVE: The Fatigue Impact Scale, FIS, is an internationally used instrument for the assessment of the impact of fatigue on Health-Related Quality of Life, HRQOL, also in patients with chronic liver diseases. In the German-speaking countries a validated instrument for measuring the impact of fatigue on patients with chronic liver diseases has not been available so far. METHODS: The German linguistic adaptation of the FIS using a forward-backward procedure was administered to 204 patients (age 52,7 +/- 13,9 years; 47 % female, 53 % male, 45 % no cirrhosis, 22 % Child's A, 15 % Child's B and 17 % Child's C cirrhosis; 53 % with chronic viral hepatitis, 32 % with alcoholic and 15 % with other liver diseases) of a secondary care hospital. The following internationally accepted instruments were used for validation: The Short Form Health Survey, SF 36, the Hospital Anxiety and Depression Scale German Version, HADS-D, and the Giessener Symptom Scale, GBB 24. Fifty patients in clinically stable situation filled out the FIS-D within 3 - 8 days. RESULTS: The acceptance of the FIS-D was high with 98 % answered items. The internal consistency of the three subscales was excellent (0.94 - 0.96), the test-retest reliability of the three subscales was good (0.72 - 0.83). The correlation coefficients with the validation instruments ranged between 0.49 and 0.80 (all p < 0 001). No differences in the FIS-D subscale scores were found in patients with and without cirrhosis and between the different Child-Pugh stages of liver cirrhosis. Patients treated with tranquilizer or antidepressants scored higher in the FIS-D than patients without this treatment (p < 0.05). CONCLUSION: The FIS-D is well accepted by patients in clinical routine care and has been shown to have good acceptance and reliability in the assessment of fatigue in chronic liver patients. PMID- 14562195 TI - Immunogenicity of an accelerated vaccination regime with a combined hepatitis a/b vaccine in patients with chronic hepatitis C. AB - OBJECTIVES: Hepatitis A (HAV) and B (HBV) vaccinations are recommended in patients with chronic liver diseases. METHODS: We prospectively investigated immunogenicity and safety of an accelerated vaccination protocol (0-7-21 days) with the combined hepatitis A/B vaccine (Twinrix(R)) versus the standard vaccination scheme (0-1-6 months) in hepatitis C virus-infected patients versus healthy volunteers. RESULTS: Local and general symptoms were mostly mild in all groups. One month after completion of the accelerated vaccination or standard vaccination, with the combined hepatitis A/B vaccine anti-HAV seroconversion rates (>33 IU/l) were 89 % and 88 % in HCV-infected patients. Initial HCV nonresponders developed protective anti-HAV antibodies in 94 % and 96 % after a booster dose. According to the anti-HBs seroprotection rate, HCV-infected patients developed protective anti-HBs titres (>10 IU/l) in 77 % and 82 % of cases one month after the accelerated and the standard vaccination scheme-at month 2 and 7, respectively. This anti-HBs seroprotection rate could even be increased to 84 % and 85 % when initial HCV-infected nonresponders where given a booster dose with the combined hepatitis A/B vaccine. Protective anti-HAV and anti-HBs titers were achieved as early as month 2 after the accelerated vaccination schedule in the majority of HCV-infected patients. Healthy subjects developed protective anti-HAV titers and anti-HBs titers in 100 % and 98 % after the accelerated and standard vaccination protocol. CONCLUSIONS: This study is the first to have demonstrated that the accelerated combined hepatitis A/B vaccination is both safe and highly immunogenic against HAV and HBV in HCV infected patients with well compensated liver disease. PMID- 14562196 TI - Spontaneous regression of an inflammatory pseudotumor of the liver. AB - Inflammatory pseudotumors (IPT) have been described for virtually every site in the body but is rare in the liver. The clinical presentation of hepatic IPT is unspecific, patients complain of fever, malaise, weight loss and often of symptoms related to a mass effect. Routine imaging procedures are not sufficient to make the diagnosis and a biopsy is necessary to differentiate IPT from neoplasms. The course of the disease is unpredictable and in most of the reported cases the patients underwent resection or were medically treated. We report a patient with a histologically proven IPT of the liver that regressed spontaneously over several months without any specific treatment. PMID- 14562197 TI - Diagnosis of hepatocellular carcinoma (HCC) in a high-risk patient by using transgastric EUS-guided fine-needle biopsy (EUS-FNA). AB - Hepatocellular carcinoma (HCC) is the most common primary malignancy arising within the liver and most often affects individuals with chronic hepatitis and/or liver cirrhosis. Patients often present at an advanced stage of disease or with poor liver function, thus limiting treatment options and resulting in a poor prognosis of the disease. Therefore, an early tissue-based diagnosis of HCC is necessary to direct further work-up and treatment. We present the case of a 70 year-old man with alcoholic cirrhosis at stage Child C, in whom a tumor nodule was found incidentally within the left lobe of the liver. Percutaneous biopsy was deemed too dangerous because a deteriorated liver function with coagulopathy was present, and a significant amount of ascites surrounded the small cirrhotic liver. To obtain a conclusive diagnosis, we performed transgastric fine-needle biopsy of the tumor under direct endosonographic guidance (EUS-FNA) without complications. Cytologic examination confirmed the presence of a well differentiated HCC. Based on this finding, super-selective CT-guided angiography and chemoembolization were subsequently performed without complications and the patient remained free of tumor relapse for the 8 months of surveillance. We conclude that EUS-guided fine-needle biopsy and cytologic examination represent a reliable alternative for tissue sampling in HCC, particularly in selected high risk patients such as those with poor liver function and coagulation disorders; this should be assessed in prospective clinical trials. PMID- 14562198 TI - [Acute appendicitis after colonoscopy: causality or coincidence?]. AB - Complications due to diagnostic colonoscopy are very rare. Perforation is the most frequent colon-related event. So far only ten cases of acute appendicitis after colonoscopy have been reported in the literature. We present the case of a 79-year-old man who developed acute appendicitis after diagnostic colonoscopy. Diagnosis was aggravated by the fact that in the patient's history an appendectomy had been performed 65 years earlier. Because of the rarity of this event it is doubtful whether the endoscopic examination was causal or coincidental. However, in cases of pain after colonoscopy, appendicitis has to be included in the differential diagnosis. PMID- 14562199 TI - [Vasoconstrictive Therapies for Bleeding Esophageal Varices and their Mechanisms of Action]. AB - Variceal bleeding is one of the most dramatic complications in gastroenterology and has a high mortality rate. Early treatment with vasoactive drugs can save lives when skilled endoscopists are not immediately available. Vasoactive drugs like terlipressin, somatostatin or octreotide are not only indicated as first choice emergency treatment, but they also increase the success rate of endoscopic treatments. Whereas the efficacy and mechanisms of action of terlipressin to arrest haemorrhage and to improve the disturbed cardiovascular situation of cirrhotic patients, including those with hepatorenal syndrome, are well documented, the efficacy and mechanisms of action of somatostatin and octreotide remain unclear and uncertain. On account of its vasoconstrictive effects on the dilated splanchnic blood vessels, terlipressin reduces blood flow into the portal vein and, thus, reduces portal venous pressure and blood flow through porto systemic shunts. As a consequence, variceal bleeding is arrested, central and arterial hypovolaemia is corrected, and activation of the renin-angiotensin aldosterone system as well as the sympathetic nervous system is reduced, leading to lower intrahepatic and intrarenal resistance. The result is an improvement of organ perfusion - including perfusion of the kidneys and the liver - as well as an improvement of the hyperdynamic cardiovascular situation and a better survival rate. Whereas terlipressin has been shown to stimulate kidney function and to prolong survival time in patients with bleeding esophageal varices as well as those with hepatorenal syndrome, no such promising effects were observed with somatostatin or octreotide. PMID- 14562200 TI - [Biologic therapy of inflammatory bowel disease]. AB - Biological therapies in inflammatory bowel disease reflect the exponential advancement in understanding the human intestinal immune system and particularly the biology of intestinal inflammation during the past decade. The better understanding of the mechanisms of inflammatory bowel disease has evolved from desriptive clinical data and genetically engineered animal models. It led to great interest in the evaluation of a variety of new therapeutic agents with novel actions. This review will discuss the mechanisms of biologicals (antibodies against pro-inflammatory cytokines, T cell antibodies, anti-inflammtory cytokines, adhesion molecule blockers, growth factors, hormones, colony stimulating factors, fusion proteins, anti-sense oligonucleotides, trefoil peptides, immunostimulatory [ISS] DNA) used in the treatment of inflammatory bowel disease and summarizes the available data on established biologic therapies as well as investigational agents and briefly touch on probiotics. Based on the data discussed, it seems that biologicals will play an important role in managing inflammatory bowel disease in the near future. PMID- 14562201 TI - [The functional relation between the gut and the liver: the newly discovered hormone relm-beta affects the action of insulin]. PMID- 14562202 TI - [PPAR-ligands: a clinically important regulatory system with multiple options in the treatment of diabetes mellitus, inflammatory bowel disease and in the chemoprevention of colon cancer?]. PMID- 14562203 TI - [ASCO Update 2003 - Highlights of the 39th Meeting of the American Society of Clinical Oncology/ASCO 2003]. PMID- 14562204 TI - [Notes of the Working Group on Ultra Sonography during 109th Meeting of the German Society of Internal Medicine, Wiesbaden 2003]. PMID- 14562207 TI - [Model Project 2004 of the DEGUM Section Gynaecology and Obstetrics]. PMID- 14562208 TI - [Sonographie von pleura und lunge]. AB - The value of ultrasound examination of the pleura and lungs remains highly underestimated to this day. While the ventilated lungs and the osseous skeleton of the thorax represent potent obstacles for ultrasound, a multitude of pathological processes of the chest wall, pleura, and lungs results in altered tissue composition, providing markedly increased access and visibility for sonographic examination. These conditions support the sonographic diagnosis of pleural and pulmonary disorders. However, the main value of pleura and lung ultrasonography is not the primary diagnosis of chest lesions but the follow up, differential diagnosis, detection of complications, such as abscesses and post embolic lung infarction, and guidance of diagnostic and therapeutic interventions in patients with pathological pleural and pulmonary findings. Punctures and drainages of fluids, e. g., haematothorax, empyema, chylothorax as well as biopsies of solid lesions can safely be performed under ultrasound-guidance. It is of special importance that pleura and lung ultrasonography, as a non-invasive method, can be repeated without discomfort or radiation exposure for the patient and is therefore valuable in the follow-up of pathological findings. Adequate interpretation of sonographic pleura and lung findings has to consider the patient's history, physical examination, chest X-ray and other results obtained by complementary imaging technologies (e. g. thoracic computed tomography). PMID- 14562209 TI - [Doppler sonographic estimation of normal values for flow velocity and resistance indices in renal arteries of healthy infants]. AB - Summary. Colour coded Duplex sonography (CDI) is an important method for the judgement of renal perfusion in infancy. By means of CDI the renal artery (RA), the segmental renal arteries (SA) and the interlobar arteries (ILA) can be differentiated. By means of pulsed Doppler sonography flow parameters in renal vessels can be measured. AIM: As flow parameters in different renal arteries are age dependent, normal values of the flow velocities and resistance indices were estimated. METHOD: We investigated 147 healthy children by CDI. Children were subdivided into 4 groups: 38 infants (< 1 year), 38 toddlers (> or = 1 year and < 6 years), 37 school children (> or = 6 and < 12 years) and 34 adolescents (> or = 12 and < 18 years). All children were investigated by computersonography with a 5 or 7,5 MHz transducer. In all children blood flow was measured in the RA, SA and ILA. From the flow profile peak systolic flow velocity (Vmax), endsystolic (Ves), enddiastolic (Ved) and time average (TAV) flow velocity as well as the resistance index (RI) were measured. RESULTS: Flow velocity and resistance indices were age dependent and related to the location within the renal vascular tree. The highest flow velocity was found in the renal artery, lowest in the ILA. In infants, Vmax in the RA was 51,5 +/- 13,4, in the SA 33 +/- 8 and in the ILA 19,5 +/- 5 cm/s. In toddlers, Vmax in the RA was 71,3 +/- 13,5, in the SA 43,6 +/- 8,5 and in the ILA 28,3 +/- 6,8 cm/s. In school age children, Vmax in the RA measured 80 +/- 18, in the SA 45,5 +/- 9,1 and in the ILA 27,9 +/- 5,3 cm/s. In adolescents, Vmax in the RA was 80,7 +/- 13,7, in the SA 46,8 +/- 11,8 and in the ILA 28 +/- 6,1 cm/s. From RA to SA the other flow velocity Ves, Ved, TAV decreased about 30 % and from SA to ILA about 30 %. Flow velocity in the different renal arteries was age dependent. The lowest flow velocity was found in newborns and infants, highest in elder school children and adolescents. The flow velocity markedly increased from infancy to early childhood whereas in elder children and adolescents a slower increase of the flow velocity could be found. Every flow velocity within the RA and AIL showed a statistically significant increase with increasing age (p < 0,0001). Resistance indices decreased from the renal artery to the interlobar arteries. The RI of infants within the RA was 0,82 +/- 0,11, within the SA 0,81 +/- 0,12 and within the AIL 0,73 +/- 0,17. In toddlers the RI within the RA was 0,71 +/- 0,08, within the SA 0,67 +/- 0,07 and within the ILA 0,65 +/- 0,08. In school age children the RI within the RA was 0,71 +/- 0,09, within the SA 0,66 +/ 0,08 and the ILA 0,58 +/- 0,10. In adolescents the RI within the RA was 0,69 +/- 0,06, within the SA 0,63 +/- 0,07 and within the ILA 0,60 +/- 0,06. Additionally, resistance indices decreased with increasing age. Statistical analysis showed a significant decrease with increasing age (p < 0,0001). Highest resistance indices could be found in early infancy and in the renal arteries, lowest resistance indices were measured in school age and adolescence and in the interlobar arteries. CONCLUSION: As flow velocity and resistance indices are age dependent and dependent on the location of the sample volume in different renal arteries, both parameters have to be considered if pathological flow parameters are measured. The determination of normal values of flow velocity and resistance indices in different renal arteries facilitate the judgement of pathologic flow parameters. For comparative controls flow measurements within the renal artery and interlobar arteries should be used. PMID- 14562210 TI - [Extracapsular spread of cervical lymph node metastases: diagnostic relevance of ultrasound examinations]. AB - OBJECTIVE: Assessment of the value of ultrasound examination in the determination of extracapsular neoplastic infiltration and soft tissue infiltration of cervical lymph nodes. MATERIAL AND METHODS: In a prospective study the results of ultrasound studies in 110 patients with squamous cell carcinoma in the head-neck region were compared with the findings of the histologic examination after neck dissection. RESULTS: Ultrasound examination showed a specificity of 81.8 % in the determination of extracapsular neoplastic infiltration. The sensitivity was only 78.6 % - this was mainly caused by microscopic extracapsular growth, which gave false-negative results. Infiltration of blood vessels were identified in all patients. There was only one false negative diagnosis of muscle infiltration. CONCLUSION: The specificity (81.8 %) and sensitivity (78.6 %) of ultrasound examinations in the diagnosis of extracapsular infiltration in patients with lymph nodes metastasis is promising. But the examiner has to keep in mind that especially microscopic extracapsular neoplastic infiltration cannot be seen in ultrasound. PMID- 14562211 TI - [Sonographic diagnosis of supracondylar fractures of the humerus]. AB - AIM: The aim of this study is to evaluate whether sonography as a primary diagnostic tool can be used for the diagnosis of supracondylar fractures of the humerus within a screening regimen. METHOD: With standardized sonography, a total of 75 children with suspected fractures of the distal humerus were investigated. The initially retrospective study (learning phase) was followed by a prospective study. All examinations were counter-checked by radiography. Supplementary to the standard sections two new sectional planes had to be added and evaluated. RESULTS: Of 65 children with fractures of the elbow, 43 had supracondylar fractures. All fractures could be visualized sonographically in the retrospective study and were diagnosed in the prospective examination. No supracondylar fracture was overlooked. Five results were falsely positive and would have been classified as undislocated supracondylar fractures type Baumann I. With the introduction of the two new sectional planes (dorsal radial and dorsal ulnar longitudinal section), rotatory deformity could be assessed. CONCLUSION: The longitudinal sections of the first compartment (standard sections) together with the two newly added dorsal radial and ulnar sections improved the diagnosis of supracondylar fractures of the humerus. The primarily unsatisfactory results could be improved by introduction of these additional sections, which are now recommended as standard sections of ultrasound application. Having acquired sufficient experience, the method can be used as a screening method for primary diagnosis and documentation of supracondylar fractures. PMID- 14562212 TI - [Ultrasound imaging of the lower urinary tract in women before and after colposuspension: a 6-month follow-up]. AB - OBJECTIVE: To evaluate the position of the bladder neck before and after open Burch colposuspension, using introital ultrasound. DESIGN: Retrospective longitudinal study using pre- and post-operative sonographic assessment of the position and function of the bladder neck. SETTING: Urogynecology units at the universities of Marburg/Gottingen and Witten/Herdecke and the DRK district hospital in Alzey, Germany. PATIENTS: 310 women undergoing open Burch colposuspension for primary genuine stress incontinence between September 1992 and December 2001. METHOD: Two-dimensional introitus sonography of the bladder neck prior to, one week and six months after surgery. RESULTS: The median age at surgery was 55 years (26 - 85). Open colposuspension lead to a 90.0 % (279/310) cure rate at 6 months with only 3.5 % (11/310) of the patients showing persistent micturation problems. A further 11.6 % (36/310) had symptoms of urgency and in 7 patients (2.3 %) a de novo urge-incontinence occurred. Post-operative bladder neck angles and movements at rest and during valsalva manoeuvre were significantly reduced while the resting bladder neck position was significantly elevated (p < 0.0001). Anatomical elevation of the bladder neck after open colposuspension varied between 2 - 39 with a median of 14.3 mm of neck elevation after one week and 9.9 mm at 6 month, respectively. Incontinence surgery lead to a significant reduction of the urethral funneling (p < 0.0001). CONCLUSION: In our series, open Burch colposuspension decreased both the bladder neck angle and the linear movement at rest and on valsalva as a result of the surgically stabilized bladder neck. Thus, our results support the hammock hypothesis that even small changes in the position of neck position are sufficient to reverse incontinence. We believe that perioperative introitus sonography is a helpful tool for the clinical assessment and documentation of not only morphological but also functional changes of the female continence organ before and after open Burch colposuspension. PMID- 14562213 TI - [Diagnosis of a Twin-Reversed-Arterial-Perfusion-Syndrome (TRAP) in the First Trimester of Pregnancy - Report on Two Cases]. AB - The Twin-Reversed-Arterial-Perfusion-Syndrome (TRAP) is a rare complication of monocygotic multiple pregnancies. It is associated with a high perinatal mortality. One twin (recipient or acardius) is perfused by the other (donour) on the basis of vascular anastomoses. Several possible therapies have been suggested. One of them is the endoscopic laser coagulation of the umbilical artery. The success of this method seems to depend on an early diagnosis of TRAP in the first trimester, when no secondary complications are to be expected. We report the diagnosis of two cases of TRAP in the 12th gestational week by transvaginal ultrasound combined with colour and pulsed Doppler. The latter facilitates the differentiation of TRAP from other pathologic conditions or malformations as well as a detailed description of the blood flow in TRAP. PMID- 14562214 TI - [Early Diagnosis of a Fetal Skeletal Dysplasia Associated with Increased Nuchal Translucency with 2D and 3D Ultrasound]. AB - We report a case of the prenatal diagnosis of a lethal skeletal dysplasia in a fetus presenting with increased nuchal translucency of 7.3 mm on a routine first trimester scan at 12 weeks of gestation. The karyotype was normal and there was no history of previous affected pregnancies. An abnormality of the fetal spine was first seen at 12 weeks of gestation, the malformation of the left hemithorax, where the processus transversi and the costal bones were replaced by a cystic structure, was detected at 13 weeks using a combination of two- and three dimensional ultrasound. In order to identify correctly the underlying problem, at 16 weeks of gestation a detailed 2D and 3D ultrasound examination was performed. 3D surface images were calculated and a combination of surface reconstruction and maximum-mode for clear delineation of the fetal skeleton was used. In this way, the vertebral defects with absence of costal bones and processus transversi in the region of the left hemithorax, which were replaced by a cystic structure causing thoracic kyphoscoliosis and pulmonal hypoplasia and were associated with marked ventriculomegaly, could be clearly demonstrated. Although fetal autopsy was not carried out, the underlying abnormalities diagnosed by ultrasound suggest Jarcho-Levin syndrome as final diagnosis, which is characterized by multiple vertebral and rib malformations. PMID- 14562215 TI - [Public health: where are we?]. PMID- 14562216 TI - [Informed consent on heparin-induced thrombocytopenia during thrombosis prophylaxis. A pilot study including 460 patients]. AB - BACKGROUND: The practicability and acceptance of written information about heparin thrombosis prophylaxis and the associated risk of heparin-induced thrombocytopenia (HIT) was evaluated in a pilot study. PATIENTS AND METHODS: All patients consecutively admitted to the department of trauma- and reconstructive surgery at the Ernst-Moritz-Arndt-University, Greifswald, Germany, with an indication for heparin thrombosis prophylaxis were give written information about thrombosis prophylaxis and the undesired drug effect HIT. After this, acceptance was evaluated using a standardized questionnaire. Primary endpoint was refusal of heparin for thrombosis prophylaxis, secondary endpoint acceptance and comprehensibility of the information. RESULTS: None of the 460 patients included in the study subsequently refused thrombosis prophylaxis with heparin. The majority welcomed the information and thought it should be given to all patients that are about to be treated with heparin. Only 0.9 % of patients judged comprehensibility of the information to be insufficient. Anticipation of imminent heparin therapy (good/very good in 90 %) and appreciation of the quality of care was not judged to be unacceptable by any patient. CONCLUSIONS: The present study demonstrates that giving information about heparin-induced thrombocytopenia during explanation of the risks and benefits of heparin thrombosis prophylaxis is feasible. This information--given in writing in our pilot study--was judged by patients to be comprehensible and necessary and did not lead to refusal of treatment. Lower incidence of HIT with use of low molecular weight heparins should be considered in the choice of drug for thrombosis prophylaxis. PMID- 14562217 TI - [Vasopressin analogue injection as ultimate measure for counteracting severe catecholamine-refractory poisoning by several vasodilators taken with suicidal intent]. AB - HISTORY AND CLINICAL FINDINGS: A 78-year-old woman was admitted to the intensive care unit 9 hours after ingestion of 2 g of isosorbitmononitrate, 430 mg of amlodipine, 250 mg of benazepril and 600 mg of mirtazapin in suicidal intent. INVESTIGATIONS: Clinical findings and invasive monitoring showed signs of a hyperdynamic hemodynamic cardiovascular failure caused by toxic vasodilatation. TREATMENT AND COURSE: Despite of primary detoxication, intravenous volume infusion with calcium gluconate, glucagon and naloxone and administration (norepinephrine up to 2 micro g/kg/min) no hemodynamic stabilization was achieved. Only when the vasopressin-analogue argipressin was given peripheral vasodilatation was overcome and hemodynamic stabilization resulted. 10 hours after discontinuing argipressin and norepinephrine the patient developed a mesenteric ischemia, and she finally died on the third day after admission. CONCLUSION: In circulatory shock due to toxic vasodilatation the use of vasopressin analogue argipressin can be helpful as an ultima therapeutic measure in catecholamine refractory shock caused by vasodilatation. Attention must be paid to overwhelming vasoconstrictor effects resulting in mesenteric ischemia. PMID- 14562218 TI - [Obligate paraneoplasias of the skin]. PMID- 14562219 TI - [Anticoagulation in stroke. Evidence-based and empirical indications]. PMID- 14562220 TI - [10 years public health in science and teaching]. PMID- 14562221 TI - [Palliative care: the demand and the realization]. PMID- 14562222 TI - [How should anticoagulants be used in recurrent, ischemic and hemorrhagic brain infarctions?]. PMID- 14562223 TI - [Do tendon lesions occur during quinolone administration?]. PMID- 14562224 TI - [The use observation--an observation of use. Does it have to be published?]. PMID- 14562225 TI - [Genetics of osteoporosis]. PMID- 14562226 TI - [Not Available]. PMID- 14562227 TI - [Not Available]. PMID- 14562228 TI - [Not Available]. PMID- 14562229 TI - [Not Available]. PMID- 14562230 TI - [Not Available]. PMID- 14562231 TI - [Not Available]. PMID- 14562233 TI - [Not Available]. PMID- 14562234 TI - [Not Available]. PMID- 14562235 TI - [Not Available]. PMID- 14562236 TI - [Not Available]. PMID- 14562238 TI - [Diverticle of the female urinary tract]. PMID- 14562237 TI - [Not Available]. PMID- 14562239 TI - [Consensus on diagnostic strategies in connatal dilatations of the urinary tract]. AB - Anomalies of the urinary tract are detected by ultrasound screening in one out of one hundred newborns. In most cases, dilatations of the upper urinary tract result from ureteropelvic junction obstruction or a primary megaureter. Especially in these cases, it is essential to differentiate between an obstruction defined as a stenosis potentially leading to progressive parenchymal damage from a dilatation without any consequences for renal function. The main criteria for assessment of urinary transport are ultrasound examinations to observe the dilatation of the collecting system, washout of the tracer during diuresis renography and lastly split renal function. Standards for investigation procedures are required to ensure comparability of treatment concepts and especially for prospective randomized studies planned for the future. A consensus concerning basic diagnostic strategies has been elaborated by a task force group consisting of members of the "Arbeitsgemeinschaft fur Padiatrische Nephrologie" in cooperation with the "Arbeitskreis Kinderurologie" in the German Society of Urology and the "Kinderurologische Arbeitsgemeinschaft" in the German Society of Pediatric Surgery with advice from leading pediatric radiologists and specialists in pediatric nuclear medicine. The main contents of this consensus are reported. PMID- 14562240 TI - [Presentation of an university]. PMID- 14562241 TI - [Colour Doppler transrectal 3D-sonography of the prostate--first experiences]. AB - INTRODUCTION: Early detection of prostate carcinoma still is problematic in spite of advantages in lab testing and sonography. This report demonstrates the new technique of colour Doppler 3-dimensional transrectal ultrasound in differential diagnosis of elevated PSA. CASE REPORTS: In 13 cases this new ultrasound technique is described and typical results are demonstrated. All cases are histologically and clinically verified. CONCLUSION: Colour Doppler 3-dimensional transrectal ultrasound is a useful new technique with specific diagnostic capabilities for differential diagnosis of an increased PSA level. Prostate carcinoma can be detected early, prostatitis can be found easily and therapeutic effects can be followed. Colour Doppler 3-dimensional transrectal ultrasound has however to prove in clinical comparative studies its specific advantages for screening of prostate carcinoma. PMID- 14562242 TI - [Control parameters for high-intensity focused ultrasound (HIFU) for tissue ablation in the ex-vivo kidney]. AB - PURPOSE: Therapeutic application of contactless thermoablation by high-intensity focused ultrasound (HIFU) demands precise physical definition of focal size and determination of control parameters. Our objective was to define the focal expansion of a new ultrasound generator and to evaluate the extent of tissue ablation under variable generator parameters in an ex vivo model. MATERIALS AND METHODS: Axial and transversal distribution of ultrasound intensity in the area of the focal point was calculated by needle hydrophone. The extent of tissue necrosis after focused ultrasound was assessed in an ex vivo porcine kidney model applying generator power up to 400 Watt and pulse duration up to 8 s. RESULTS: The measurement of field distribution revealed a physical focal size of 32 x 4 mm. Sharp demarcation between coagulation necrosis and intact tissue was observed in our tissue model. Lesion size was kept under control by variation of both generator power and impulse duration. At a constant impulse duration of 2 s, generator power of 100 W remained below the threshold doses for induction of a reproducible lesion. An increase in power up to 200 W and 400 W, respectively, induced lesions with diameters up to 11.2 x 3 mm. Constant total energy (generator power x impulse duration) led to a larger lesion size under higher generator power. CONCLUSION: It is possible to induce sharply demarcated, reproducible thermonecrosis, which can be regulated by generator power and impulse duration, by means of a cylindrical piezo element with a paraboloid reflector at a focal distance of 10 cm. The variation of generator power was an especially suitable control parameter for the inducement of a defined lesion size. PMID- 14562243 TI - [Complex malformation: cake kidney with concomitant sacral agenesis]. AB - Ectopic kidneys are frequently associated with primary renal dysplasia or a disturbance of urine transport. Sacral agenesis is defined by the absence of two or more bodies of the lower vertebral and is often associated with a neurogenic bladder dysfunction. A 5-year-old boy with sacral agenesis and a right-sided cake kidney, presented with progredient renal failure caused by recurrent urinary tract infections, incomplete bladder emptying and vesicorenal reflux. After extensive diagnostics, the anatomical situation was explored by laparotomy to find a solution for this complex situation. Despite modern diagnostic tools, the preoperatively estimated renal function of the cake kidney was incorrect. After resecting of the right collecting system and refluxing megaureter out of the cake kidney, anti-refluxive implantation of the left ureter into the bladder was performed and the megacystis was treated by detrusor duplication. After three years, the now 8-year-old boy voids residual free without any signs of urinary tract infection. Renal function and proteinuria have improved. The only medication required is nifedipine (20 mg twice a day) for treatment of the renal hypertension. PMID- 14562244 TI - [Urethral diverticulum in the female. Case report, controversy and treatment]. AB - INTRODUCTION: Urethral diverticulum is a rare cause of recurrent urinary tract infection in women. CASE REPORT: We report a single case and discuss controversially the different surgical treatment options. The safety of the Spence technique of saucerisation is particularly emphasized. CONCLUSION: In our opinion the Spence saucerisation technique is the treatment of choice in all cases of urethral diverticulum in the female. PMID- 14562245 TI - [Not Available]. PMID- 14562246 TI - An introduction to Staphylococcus aureus, and techniques for identifying and quantifying S. aureus adhesins in relation to adhesion to biomaterials: review. AB - The ability of Staphylococcus aureus to adhere to the extracellular matrix and plasma proteins deposited on biomaterials is a significant factor in the pathogenesis of orthopaedic-device related infections. S. aureus possesses many adhesion proteins on its surface, but it is not known how they interact with each other to form stable interactions with the substrate. A novel method was developed for extracting adhesins from the S. aureus cell wall, which could then be further analysed. The protocol involves using a FastPrep instrument to mechanically disrupt the cell walls resulting in native cell walls. Ionically and covalently bound proteins were then solubilised using sodium dodecyl sulphate (SDS) and lysostaphin, respectively. Western blot analysis of covalently bound proteins using anti-protein A and anti-clumping factor A sera showed that S. aureus produces most surface proteins in early growth, and less in post exponential and stationary growth. Immuno-gold labelling of protein A, and clumping factor A was observed all over the bacteria and showed no distinct surface distribution pattern. However, this labelling showed expression of surface associated proteins varied in a growth-phase dependent and cell-density dependent manner. PMID- 14562248 TI - Fabrication of highly porous scaffold materials based on functionalized oligolactides and preliminary results on their use in bone tissue engineering. AB - Tissue engineering offers a promising new approach to repair bone defects. Its practical realisation is connected with the development of suitable scaffold materials. In the present work, functionalized oligolactides have been prepared and used as macromers for the scaffold fabrication The developed fabrication process leads to highly porous scaffolds, available in various shapes and sizes, with an open inter-connective pore structure and porosities up to 90%. Degradable or even osteoconductive components as well as biocompatible co-monomers can be used as additives to modulate the scaffold properties. Under in vitro conditions, the scaffolds exhibit a continuous degradation with varying degradation rates depending on their material composition. In vitro studies on the cultivation of osteoblasts on the scaffolds were performed and revealed their excellent biocompatibility. Cell growth on the scaffold surfaces and inside the scaffolds, formation of extracellular matrix and starting mineralization were detected by microscopical and histological analyses. Based on these results the developed materials are well-suited candidates for the design of tailor-made matrices in bone tissue engineering PMID- 14562247 TI - Behaviour of moderately differentiated osteoblast-like cells cultured in contact with bioactive glasses. AB - Bioactive glasses have been shown to stimulate osteogenesis both in vivo and in vitro. However, the molecular mechanisms underlying this process are still poorly understood. In this study, we have investigated the behaviour of osteoblast-like cells (MG63), cultured in the presence of bioglass particles. Three types of granules were used: 45S5 bioactive glass, 45S5 granules preincubated in tris buffer and 60S non-reactive glass, used as control. Phase contrast microscopy permitted step-by-step visualization of cell cultures in contact with the particles. Ultrastructural observations of undecalcified sections revealed direct contacts of the cells and an electron-dense layer located at the periphery of the material. Protein synthesis was evaluated biochemically and showed a gradual increase throughout the culture time in the three types of cultures. Alkaline phosphatase was detected in situ, in clusters of packed cells either in contact with the material or in the background cell layer. Semi-quantitative RT-PCR analysis of the main osteoblastic markers showed that gene expression was maintained in all three cultures. The fact that osteocalcin was not detected, supports the fact that the MG63 cell line is composed of less differentiated osteogenic cells rather than mature osteoblasts. We also demonstrated for the first time in this cell line, the expression of Msx-2, Dlx-3 and Dlx-7 homeogenes, known to regulate in vivo foetal skeletogenesis as well as adult skeletal regeneration. However, no significant differences could be recognised in the expression pattern of bone markers between the three types of cultures. Yet these preliminary results indicate that bioactive glasses provided a suitable environment for the growth and proliferation of osteoblasts in vitro, since no drastic changes in phenotype expression of pre-osteoblasts was noted. PMID- 14562249 TI - Novel aspects to the structure of rabbit articular cartilage. AB - Applying cryo and modified chemical preparation techniques, mainly for scanning electron microscopy, revealed entirely new aspects to the structure of the radial zone of rabbit tibial plateau articular cartilage. The aggrecan component of the extracellular matrix was contained radially in columns, each with a diameter of 1 3 mm, by a tightly packed matrix of collagen fibrils. The collagen fibrils were arranged radially, some straight and others in an opposed spiral arrangement, with regularly repeating patterns. This organization existed in the regions surrounding the columns of chondrocytes, known as chondrons. The load bearing property of the tissue was explained by the directed flow and containment of the interstitial fluid, modulated by the protein-carbohydrate complexes, along these collagen bounded tubular structures. The reason why such a structure has not been described previously may be that it is not retained by aldehyde fixation followed by dehydration, the method commonly used for tissue preparation for electron microscopy. PMID- 14562250 TI - Fibroblast and osteoblast adhesion and morphology on calcium phosphate surfaces. AB - Implant loosening in bone fixation is an unresolved complication associated with internal fixation. It is generally accepted that this problem can be overcome by modifying the implant/bone interface for improved osseointegration. This is achieved, in part, by hydroxyapatite (HA) or tricalcium phosphate coatings. Unfortunately, the benefits of these coatings are constrained by not only the generally low strength of their adhesion to the implant surface but also the limited cohesion within their layers. Anodic Plasma-chemical treatment (APC) has been developed to incorporate electrolytes and produce coatings with various microtopographies and strong adhesion to implants. In this in vitro study fibroblast and osteoblast morphologies and adhesion to various substrates were evaluated using qualitative and quantitative methods. The substrates were Thermanox plastic and commercially pure titanium. The latter were surface-treated using several different methods: conventional anodisation, plasma spraying of HA and anodic plasma-chemical (APC) treatment in an electrolyte solution containing either calcium and phosphate (APC-CaP) or phosphoric acid (APC-P). Both osteoblasts and fibroblasts showed extensive cell spreading, total cell area and greatest amount of adhesion, with defined adhesion patterns on the Thermanox plastic, anodised titanium, and the two APC-CaP substrates. With fibroblasts, almost no cell spreading and very low adhesion, was observed in cells cultured on the APC-P and HA surfaces. The extent of cell spreading correlated with the area of focal adhesions as assessed by the amount of vinculin labelling. The Thermanox plastic, anodised titanium, and the two APC-CaP substrates were the most cytocompatible substrates with regard to this in vitro evaluation. PMID- 14562251 TI - Differentiation of rat osteoblast-like cells in monolayer and micromass cultures. AB - During intramembranous bone formation, preosteoblasts condense, differentiate into osteoblasts and deposit bone matrix. We compared the differentiation process of rat calvarial osteoblast-like cells inoculated as micromasses, which mimic the in vivo condensation process, with cells inoculated as monolayers. The cells were analysed morphologically at 1,2 and 3 weeks by light microscopy (alkaline phosphatase activity, mineralization), by transmission electron microscopy, and biochemically (collagen typing, alkaline phosphatase activity, protein and DNA content). The cells inoculated as monolayers formed alkaline phosphatase positive and mineralized nodules during the culture period. The cells inoculated as a micromass formed a large mineralized area consisting of smaller nodules. The ultrastructure of the cells in both culture systems showed the typical features of osteoblasts and osteocytes. The main difference between monolayer and micromass cultures was found after 1 week in culture. The cells inoculated as a micromass formed a multilayer of cells. The cytoplasm contained rER, mitochondria, vesicles and ribosomes. There were abundant collagen fibrils in membrane folds and in the extracellular matrix. This was in contrast to the cells in monolayer culture which showed hardly any collagen fibrils in the extracellular matrix. The promotion of the differentiation was also confirmed by biochemical data showing that the DNA content was lower in the micromass than in the monolayer cultures during the culture period. These results show that micromass, as compared to monolayer, culture promotes the differentiation of rat osteoblast-like cells in vitro. PMID- 14562252 TI - Cell fitting to adhesive surfaces: A prerequisite to firm attachment and subsequent events. AB - Cell adhesion usually involves extensive shape reorganization. This process is important because i) it is required for efficient cross-linking of interacting surfaces by adhesion receptors the length of which does not exceed several tens of nanometers and ii) it influences subsequent cell differentiation and activation. This review focuses on the initial phase of cell deformation, preceding the extensive reorganization process known as spreading. This first phase includes local flattening at the micrometer scale and membrane alignment at the nanometer level, resulting in fitting of the cell to an adhesive surface. Three main points are considered. First, experimental methods available to study cell apposition to a surface are described, with an emphasis on interference reflection microscopy. Second, selected experimental evidence is presented to show that there is a quantitative relationship between "adhesiveness" and "contact extension", and some theoretical models aimed at relating these parameters are briefly sketched. Third, experimental data on the kinetics of initial contact extension are described and possible mechanisms for driving this extension are discussed, including nonspecific forces, receptor-mediated interactions, active cell movements or passive membrane fluctuations. It is concluded that both passive physical phenomena and random active cell movements are possible candidates for the initial triggering of contact extension. PMID- 14562253 TI - Phenotypic modulation of human articular chondrocytes by bistratene A. AB - Chondrocytes undergo phenotypic alterations following extended periods in monolayer culture, i.e., they become bipolar and flattened, proliferate, and synthesise type I as opposed to type II collagen. This process has been termed chondrocyte dedifferentiation. Bistratene A is a macrolide polyether that specifically activates the delta isoform of protein kinase C (PKCdelta) in some cell types. Here, we show that dedifferentiated human articular chondrocytes became rounded and underwent cell growth arrest after treatment with bistratene A. In addition, bistratene A-treated chondrocytes became more immunopositive for type II collagen, but less immunopositive for type I collagen. These phenotypic changes were associated with a prior and extensive disruption of actin microfilaments and translocation of PKCdelta to the nuclear membrane. Concurrent treatments of chondrocytes with a specific inhibitor of PKCdelta, rottlerin, partially blocked the morphological effects of bistratene A. PMID- 14562254 TI - Application of X-ray microanalysis to the study of cell physiology in cells attached to biomaterials. AB - X-ray microanalysis is commonly applied in biomaterials research to study changes in biomaterial composition, calcifications, or to identify particulates in tissue that has been in contact with biomaterials. Studies where the effect of biomaterials on the naturally occurring elements in the cells are studied are rare. Exposure to or contact with biomaterials may give rise to several cellular reactions with characteristic changes in elemental patterns. Experimental systems in which cultured cells are exposed to biomaterials, and the resulting changes in elemental content measured by X-ray microanalysis are presented. Proliferation results in an increase in Mg, P and K. Injury and necrosis result in increased Na, Cl and Ca, and decreased K and Mg. The Na/K ratio is a sensitive indicator of cell damage. Apoptosis results in increased Na/K and an increase P/S ratios. Mechanical effects of biomaterials may cause changes in cell-shape or cell volume that may result in volume-regulating ion fluxes, e.g., of chloride, accompanied by efflux of potassium. The multi-element detection capability of X-ray microanalysis makes it particularly suitable to detect these characteristic patterns of changes. The technique can also be used to define sub populations of cells that differ with respect to their reactions to external stimuli. PMID- 14562255 TI - Chloride channels and the reactions of cells to topography. AB - The reactions of rat epitenon cells to substratum topography on the micrometric and nanometric scale such as groove-ridge structures include cell extension, elongation and orientation reactions. In this paper we report that stretch sensitive chloride channels may be involved in the earliest stages of these reactions in epitenon fibroblast-like cells. We report that rat epitenon-cells can develop appreciable lateral mechanical tension that could stretch both the force generating cells themselves and those nearby. We show that cells in medium in which more than 80% of the chloride has been replaced by nitrate show little reaction to topography. Spreading of the cells takes place but is much reduced along the direction of the groove-ridge topography but enhanced across the topography. The chloride channel inhibitors NPPB (5-Nitro-2- (3phenylpropylamino) benzoicacid) 4,4'-disothiocyanostilbene-2, 2' sulphonic acid (DIDS) and Chlorotoxin produce similar results which are further accentuated when these inhibitors are presented in low chloride medium. An antibody against ClC3, which has close homology to ClC5/6 also, blocked reaction to topography. These treatments have no significant effect on cell spreading on planar surfaces nor do they lead to changes in internal pH in the cells. There is a slight inhibition of rates of cell movement. Experiments using antisense oligoribonucleotides to ClC-5 or ClC-6 channel m-RNA also inhibit topographic reactions, which provides further confirmation of the hypothesis. Since the ClC-3,4 and 5 share considerable sequence similarities in the genes and in their proteins it has not been possible to make an unambigous determination of which precise chloride channel(s) is (are) involved. PMID- 14562256 TI - Structure and substitutions in fluorapatite. AB - Fluorapatite, Ca10(PO4)6F2. is a widely spread form of calcium phosphate present particularly in biological material. Human hard tissues contain crystals structurally related to apatite. Fluoride can be found in various natural sources and is also used for its beneficial action in caries prevention. Fluorapatite belongs to the spatial group P6(3/m) (C(6h)2) and consists of 3 ions: F-, Ca2+, PO4(3-). In the present paper, we have carried out a crystallographic study of the fluorapatite structure and of the changes induced by the substitutions. The fluorapatite structure and the presence of a large number of ionic bonds make fluorapatite a very suitable host for many substituents, some of them harmless for the human organism, some not. According to the substitution site, we can describe four types of substitution. The F- substitution, also called Type A substitution, is the main one, and the best known. Only the Ca2+ substitution implies changes in the crystal structure. However, some questions remain, in particular for the PO4(3-) substitution, which is the main substitution present in the biological calcium phosphates. PMID- 14562257 TI - Interaction between Saccharomyces cerevisiae and chrysotile. AB - The interaction between Saccharomyces cerevisiae and chrysotile fibers was studied by scanning electron microscopy. The yeast cells adhere preferentially to the fibrils. In the extreme case, all the adhered fibrils were broken, resulting in a complete coverage of the surface. The chrysotile covered cells showed less buds, but retained metabolic capacities, and were fully active in fermentation experiments after one year. The interaction degree was depending on contact time and adhesion medium. The longer the contact period, the stronger the interaction between the cells and the fibers. Cells adhered in water show poor entrapment after short contact time, but were highly entrapped after longer periods and did not show any agglomerates. Cells adhered in the presence of nutrients showed a lower entrapment and a higher degree of cellular growth. PMID- 14562258 TI - Carbon materials in the treatment of soft and hard tissue injuries. AB - Carbon-based implant materials are of interest because they are well accepted by the biological environment. Carbon fibrous materials developed in the Department of Special Ceramics of the University of Mining and Metallurgy in Cracow were tested in in vivo studies to determine their influence on the living body. For comparative purposes, different carbon fibers were prepared and subjected to different surface modifications. Carbon materials prepared in the form of braids were implanted in subcutaneous tissue of rabbits and into skeletal muscle of rats. Carbon fabrics were examined as scaffolds in reconstruction of bone defects. The present study examined the synthesis-structure-property relationships of fibrous carbon samples with respect to the tissue response. It was shown that the tissue response depends on the form of the material form, the degree of order of the crystallites, the surface state and microstructural parameters. Carbon fibers with higher crystallinity and a better-organized graphite structure were assimilated by the body with more difficulty and small particles coming from these materials were found in the regional lymph nodes. Low carbonized carbon fibers (small crystallite size) underwent partial fragmentation and reacted with the biological environment by being gradually resorbed in the implantation site. The presence of acidic groups on the surface of the carbon fibers enhanced phagocytosis of the carbon material by macrophages. Depending on the surface state of carbon fibers different rates of bone wound healing were observed. PMID- 14562259 TI - Influence of cell isolation, cell culture density, and cell nutrition on differentiation of rat calvarial osteoblast-like cells in vitro. AB - The effects of various cell isolation procedures, growth media and the cell culture density on the in vitro differentiation of neonatal rat calvarial osteoblast-like cells were investigated. Cells were isolated by enzymatic treatment, or after explant culture and inoculated as a monolayer or micromass in serum containing BGJb, or Dulbecco's Modified Eagle Medium (DMEM). The cells were kept for up to 3 weeks in culture and were then characterized, both morphologically and biochemically. The isolation technique appeared to have no effect on the differentiation process. The calvaria could be used several times as explant cultures for a reliable source of differentiating osteoblast-like cells. The cultures kept in DMEM had a significantly higher DNA content, but significantly less alkaline phosphatase activity (ALP) per DNA and protein per DNA content than the BGJb cultures. Monolayer cultures had a significantly higher DNA content than the micromass cultures, in both growth media. Furthermore, the micromass culture had a significantly higher ALP per DNA than monolayer cultures at 1 week. The morphology of all cell cultures at 3 weeks reflected the biochemical results. Only the cells grown in BGJb formed abundant ALP positive and mineralized nodules in monolayer cultures. In contrast, cells grown as micromasses formed a dense calcified area, independently of the growth medium used. DMEM promoted the proliferation, whereas BGJb stimulated the differentiation of osteoblast-like cells in monolayer cultures. Micromass cultures were less sensitive to nutritional conditions than monolayer cultures and promoted the differentiation of osteoblast-like cells. PMID- 14562260 TI - The influence of pore geometry in cp Ti-implants--a cell culture investigation. AB - Biocompatibility testing of differently structured titanium implants was performed using an in vitro test system of a newly established human fetal osteoblastic cell line (hFOB 1.19). Cell adhesion of osteoblastic cells on the different porous geometries and the suitability of a copper vapor laser system for surface structuring was tested with the following parameters: cell-number, cell viability, alkaline phosphatase expression. The analysis of the cell culture results demonstrated that 25 microm and 200 microm porous geometries showed similar or even better results than the negative control of polystyrene; there was no sign of toxic effects. However, the 100 microm porous geometry showed an impressive negative influence on the calculated parameters. The reason for this effect is unclear. The series with 50 microm, 300 microm, 400 microm and 500 microm showed a comparable, intermediate effect on the cell culture with respect to the different parameters. However, the results were worse than with the 25 and 200 microm porous geometry. In conclusion, the 25 microm and 200 microm porous geometry seems to have the most positive effect on the human osteoblastic cell line hFOB 1.19. PMID- 14562261 TI - Animal models of osteoporosis--necessity and limitations. AB - There is a great need to further characterise the available animal models for postmenopausal osteoporosis, for the understanding of the pathogenesis of the disease, investigation of new therapies (e.g. selective estrogen receptor modulators (SERMs)) and evaluation of prosthetic devices in osteoporotic bone. Animal models that have been used in the past include non-human primates, dogs, cats, rodents, rabbits, guinea pigs and minipigs, all of which have advantages and disadvantages. Sheep are a promising model for various reasons: they are docile, easy to handle and house, relatively inexpensive, available in large numbers, spontaneously ovulate, and the sheep's bones are large enough to evaluate orthopaedic implants. Most animal models have used females and osteoporosis in the male has been largely ignored. Recently, interest in development of appropriate prosthetic devices which would stimulate osseointegration into osteoporotic, appendicular, axial and mandibular bone has intensified. Augmentation of osteopenic lumbar vertebrae with bioactive ceramics (vertebroplasty) is another area that will require testing in the appropriate animal model. Using experimental animal models for the study of these different facets of osteoporosis minimizes some of the difficulties associated with studying the disease in humans, namely time and behavioral variability among test subjects. New experimental drug therapies and orthopaedic implants can potentially be tested on large numbers of animals subjected to a level of experimental control impossible in human clinical research. PMID- 14562262 TI - Cell reactions with biomaterials: the microscopies. AB - The methods and results of optical microscopy that can be used to observe cell reactions to biomaterials are Interference Reflection Microscopy (IRM), Total Internal Reflection Fluorescence Microscopy (TIRFM), Surface Plasmon Resonance Microscopy (SPRM) and Forster Resonance Energy Transfer Microscopy (FRETM) and Standing Wave Fluorescence Microscopy. The last three are new developments, which have not yet been fully perfected. TIRFM and SPRM are evanescent wave methods. The physics of these methods depend upon optical phenomena at interfaces. All these methods give information on the dimensions of the gap between cell and the substratum to which it is adhering and thus are especially suited to work with biomaterials. IRM and FRETM can be used on opaque surfaces though image interpretation is especially difficult for IRM on a reflecting opaque surface. These methods are compared with several electron microscopical methods for studying cell adhesion to substrata. These methods all yield fairly consistent results and show that the cell to substratum distance on many materials is in the range 5 to 30 nm. The area of contact relative to the total projected area of the cell may vary from a few per cent to close to 100% depending on the cell type and substratum. These methods show that those discrete contact areas well known as focal contacts are frequently present. The results of FRETM suggest that the separation from the substratum even in a focal contact is about 5 nm. PMID- 14562263 TI - Electron diffraction and high resolution transmission electron microscopy in the characterization of calcium phosphate precipitation from aqueous solutions under biomineralization conditions. AB - Calcium phosphate precipitation obtained from aqueous solutions at room and body temperature and pH 5.5-7.5 were investigated by high-resolution transmission electron microscopy (HRTEM), transmission electron diffraction, scanning electron microscopy (SEM) and X-ray diffraction (XRD). Supersaturated solutions of calcium phosphates were prepared by different methods of mixing of the stock solutions: diffusion-controlled mixing in space, convection-controlled mixing on earth and forced mixing on earth and with typical physiological parameters (pH and temperature). Concentrations of the stock solutions, rate of solution mixing and duration of precipitation influence very strongly the chemical composition of the precipitation, the phase composition of individual crystals, their sizes, morphology and structure. Microdiffraction and HRTEM techniques showed an incontestable advantage on other techniques like SEM and XRD in the investigation of small particles and mixtures of calcium phosphates (hydroxyapatite and octacalcium phosphate) with different proportions. PMID- 14562264 TI - Precision of high-resolution dual energy X-ray absorptiometry of bone mineral status and body composition in small animal models. AB - The purpose of this study was to analyze the in situ precision (reproducibility) of bone mineral and body composition measurements in mice of different body weights and rats, using a high-resolution DXA (dual energy X-ray absorptiometry) scanner. We examined 48 NMRI mice weighing approximately 10 to 60 g, and 10 rats weighing approximately 140 g. Four repeated measurements were obtained on different days. In mice, the standard deviations of repeated measurements ranged from 2.5 to 242 mg for bone mineral content (BMC), from 0.16 to 3.74 g for fat, and from 0.40 to 4.21 g for lean mass. The coefficient of variation in percent (CV%) for BMC/BMD (bone mineral density) was highest in the 10 g mice (12.8% / 4.9%) and lowest in the 40 g mice (3.5% /1.7%). In rats, it was 2.5 /1.2% in the lower extremity, 7.1/3.0 % in the spine, 5.7/2.0 % in the femur, and 3.6%/2.1% in the tibia. The CV% for fat and lean mass in mice was higher than for BMC. The study demonstrates good precision of bone mineral and moderate precision of body composition measurements in small animals, using a high-resolution DXA system. The technique can be used for testing the efficacy of drugs in small animal models, for mutagenesis screens, and for the phenotypic characterization of transgenic mice. PMID- 14562265 TI - Characterizing fretting particles by analysis of SEM images. AB - Detailed surface characteristics of the particles produced during fretting may well be significant in determining their biological effects. Apart from a broad size determination very few attempts have been made at devising means of describing profile textures of particles. An approach is presented for describing the nature of particle shape and surface texture, as detected on the particle profile. Careful processing and analysis of the digitised image enabled both the sizes of micro-projections and their relative numbers to be determined. Such processing of images of a large number of particles generated a considerable amount of data. An artificial neural network was used to categorise the data and to compare the nature of fretting particles generated by titanium, titanium molybdenum and stainless steel. Although showing a tendency towards a spherical form, all three metals produced different results, with titanium showing the greatest diversity of textures and sizes, steel the least. PMID- 14562266 TI - A combined atomic force microscopy and nanoindentation technique to investigate the elastic properties of bone structural units. AB - To our knowledge, this study applied for the first time a recently developed combination of atomic force microscopy (AFM) and nanoindentation on trabecular and compact bone tissue. The major aim was to check the advantage of the available AFM-mode over the conventionally used optical microscope. First, we investigated if removal of the water content helped to prevent enzymatic degradation of the bone tissue and preserve its mechanical properties during a week. After the positive issue of this test, we quantified the intrinsic mechanical properties of single bone structural units (BSU). Bone specimens were obtained from the femoral neck of an 86 year old female. Four BSU were randomly selected and tested each with 24 indents of 5 mN maximum force. The available AFM mode proved to be a very useful tool for surface characterization and precise selection of the indentation area. The elastic modulus ranged from 18 -/+ 1.7 GPa for a BSU of compact bone to 22.5 -/+ 3.1 GPa for a BSU of trabecular bone. Hardness showed values between 0.6 -/+ 0.11 GPa for compact bone and 1.1 -/+ 0.17 GPa for trabecular bone. The results suggest that the micromechanics of bone tissue may also be described as an assembly of distinct structural units with rather homogeneous material properties. PMID- 14562267 TI - The bisphosphonate zoledronate prevents vertebral bone loss in mature estrogen deficient rats as assessed by micro-computed tomography. AB - The effect of long-term treatment with the bisphosphonate zoledronate on vertebral bone architecture was investigated in estrogen-deficient mature rats. 4 month-old rats were ovariectomized and development of cancellous osteopenia was assessed after 1 year. The change of bone architectural parameters was determined with a microtomographic instrument of high resolution. After 1 year of estrogen deficiency, animals lost 55% of vertebral trabecular bone in comparison to sham operated control animals. Trabecular number (Tb.N) and trabecular thickness (Tb.Th) were significantly reduced in ovariectomized animals, whereas trabecular separation (Tb.Sp), bone surface to volume fraction (BS/BV) and trabecular bone pattern factor (TBPf) were significantly increased, indicating a loss of architectural integrity throughout the vertebral body. 3 groups of animals were treated subcutaneously with zoledronate for 1 year with 0.3, 1.5 and 7.5 microgram/kg/week to inhibit osteoclastic bone degradation. Administration started immediately after ovariectomy and treatment dose-dependently prevented the architectural bone deterioration and completely suppressed the effects of estrogen deficiency at the higher doses. The results show that microtomographic determination of static morphometric parameters can be used to quantitate the effects of drugs on vertebral bone architecture in small laboratory animals and that zoledronate is highly effective in this rat model. PMID- 14562268 TI - Differential roles for small leucine-rich proteoglycans in bone formation. AB - This paper reviews our current state of knowledge of the roles the small leucine rich proteoglycans (SLRPs) play in the formation of connective tissue and mineralised tissue matrices. Both, the SLRPs biglycan and decorin are highly expressed in extracellular bone matrix and there is now substantial evidence to support an increasing role for biglycan and decorin in influencing bone cell differentiation and proliferative activity. In addition decorin and biglycan have been implicated in regulating mineral deposition and crystal morphology, whilst decorin has also roles in organic matrix assembly. In order to further assess the role of these SLRPs during bone formation we have initiated studies investigating primary bone cell culture models from rats (bone marrow stromal cells, and bone cells from alveolar bone explants), and identified periods relating to cell proliferation, organic matrix deposition, remodeling of the osteoid, and mineral deposition. Analysis of mRNA levels and the nature of the proteoglycan demonstrated that dermatan sulphate substituted biglycan was expressed during phases relating to cell proliferation, ceased at early matrix deposition, and then biglycan was re-expressed at the onset of mineralisation, but was conjugated to chondroitin sulphate. Decorin was expressed later than biglycan, was associated with early matrix deposition, but then continued to the mineralisation stages. Again, dermatan sulphate-decorin prevailed earlier within osteoid matrix, whilst chondroitin sulphate-decorin predominated later within the mineralizing matrix. The nature of the GAG chain conjugated to SLRP and the timing of its expression would seem to dictate the functions biglycan and decorin play in bone formation. PMID- 14562269 TI - A bone fluid flow hypothesis for muscle pump-driven capillary filtration: II. Proposed role for exercise in erodible scaffold implant incorporation. AB - A model is presented for enhancement of fluid flow through bone matrix and any porous tissue engineering scaffold implanted within it. The mechanism of enhancement is the skeletal muscle pump in compartments adjacent to the bone. Pressure waves from muscle pump contractions aided by increased blood pressure during exercise coupled with temporary occlusion of arteries leading to and veins from the bone, increase hydraulic pressure in cortical bone capillaries so as to amplify capillary filtration. It is proposed that capillary filtration increase is sufficiently convective to contribute to bone fluid flow and associated percolation through tissue engineered scaffold matrix implants. Importance of this contribution is its relative role in maintaining seeded cells in bioreactor scaffolds. Validation of the hypothesis starts at a minimum level of demonstrating that capillary filtration is convective. At a maximum level confirmation of the hypothesis requires demonstration that capillary filtration based interstitial flow is sufficient to stimulate not only host bone cells (as proposed in part I of the hypothesis) but bioreactor-seeded cells as well. Preliminary data is presented supporting the prediction that skeletal muscle contraction generates convective capillary filtration. PMID- 14562270 TI - Making tissue engineering scaffolds work. Review: the application of solid freeform fabrication technology to the production of tissue engineering scaffolds. AB - Tissue engineering is a new and exciting technique which has the potential to create tissues and organs de novo. It involves the in vitro seeding and attachment of human cells onto a scaffold. These cells then proliferate, migrate and differentiate into the specific tissue while secreting the extracellular matrix components required to create the tissue. It is evident, therefore, that the choice of scaffold is crucial to enable the cells to behave in the required manner to produce tissues and organs of the desired shape and size. Current scaffolds, made by conventional scaffold fabrication techniques, are generally foams of synthetic polymers. The cells do not necessarily recognise such surfaces, and most importantly cells cannot migrate more than 500 microm from the surface. The lack of oxygen and nutrient supply governs this depth. Solid freeform fabrication (SFF) uses layer-manufacturing strategies to create physical objects directly from computer-generated models. It can improve current scaffold design by controlling scaffold parameters such as pore size, porosity and pore distribution, as well as incorporating an artificial vascular system, thereby increasing the mass transport of oxygen and nutrients into the interior of the scaffold and supporting cellular growth in that region. Several SFF systems have produced tissue engineering scaffolds with this concept in mind which will be the main focus of this review. We are developing scaffolds from collagen and with an internal vascular architecture using SFF. Collagen has major advantages as it provides a favourable surface for cellular attachment. The vascular system allows for the supply of nutrients and oxygen throughout the scaffold. The future of tissue engineering scaffolds is intertwined with SFF technologies. PMID- 14562271 TI - Osteogenesis imperfecta--clinical and molecular diversity. AB - Osteogenesis imperfecta is a heritable disorder of bone formation resulting in low bone mass and a propensity to fracture. It exhibits a broad range of clinical severity, ranging from multiple fracturing in utero and perinatal death to normal adult stature and a low fracture incidence. The disorder is currently classified into seven types based on differences in clinical presentation and bone architecture. Mutation in one of the type I collagen genes is commonly associated with osteogenesis imperfecta, but is not a prerequisite for the diagnosis. Indeed, the newer forms of osteogenesis imperfecta (types V, VI and VII) are not associated with type I collagen gene defects. Amongst the type I collagen gene mutations that can occur, missense base substitutions involving glycine codons in the exons encoding the central triple-helix forming domain predominate. Such mutations can occur in all the classical forms of osteogenesis imperfecta (types I-IV), but genotype/phenotype correlations are complex and often unpredictable. Treatment of osteogenesis imperfecta by bisphosphonate therapy can improve bone mass in all types of the disorder, and while not being a cure for the disorder does improve the quality of life of the patient. PMID- 14562272 TI - Development of a mechanical testing and loading system for trabecular bone studies for long term culture. AB - A highly accurate (+/-3%) mechanical loading and measurement system combined with a trabecular bone diffusion culture-loading chamber has been developed, which provides the ability to study trabecular bone (and possibly) cartilage under controlled culture and loading conditions over long periods of time. The loading device has been designed to work in two main modes, either to apply a specific compressive strain to a trabecular bone cylinder or to apply a specific force and measure the resulting deformation. Presently, precisely machined bone cylinders can be loaded at frequencies between 0.1 Hz to 50 Hz and amplitudes over 7,000 microepsilon. The system allows accurate measurement of many mechanical properties of the tissue in real time, including visco-elastic properties. This paper describes the technical components, reproducibility, precision, and the calibration procedures of the loading system. Data on long term culture and mechanical responses to different loading patterns will be published separately. PMID- 14562273 TI - Platelet adhesion studies on dipyridamole coated polyurethane surfaces. AB - Surface modification of polyurethanes (PUs) by covalent attachment of dipyridamole (Persantin) is known to reduce adherence of blood platelets upon exposure to human platelet rich plasma (PRP). This effect was investigated in further detail. First platelet adhesion under static conditions was studied with four different biomaterial surfaces: untreated PU, PU immobilised with conjugate molecule 1, PU immobilised with conjugate molecule 2, and PU immobilised with conjugate molecule 3. In PU immobilised with 1 dipyridamole is directly linked to the surface, in PU immobilised with 2 there is a short hydrophilic spacer chain in between the surface and the dipyridamole, while conjugate molecule 3 is merely the spacer chain. Scanning electron microscopy (SEM) was used to characterise platelet adhesion from human PRP under static conditions, and fluorescence imaging microscopy was used to study platelet adhesion from whole blood under flow. SEM experiments encompassed both density measurements and analysis of the morphology of adherent platelets. In the static experiments the surface immobilised with 2 showed the lowest platelet adherence. No difference between the three modified surfaces emerged from the flow experiments. The surfaces were also incubated with washed blood platelets and labeled with Oregon-Green Annexin V. No capture of Oregon-Green Annexin V was seen, implying that the adhered platelets did not expose any phosphatidyl serine at their exterior surface. PMID- 14562274 TI - Haemocompatibility evaluation of DLC- and SiC-coated surfaces. AB - Diamond-like carbon (DLC) and silicon carbide (SiC) coatings are attractive because of low friction coefficient, high hardness, chemical inertness and smooth finish, which they provide to biomedical devices. Silicon wafers (Si(waf)) and silicone rubber (Si(rub)) plates were coated using plasma-enhanced chemical vapour deposition (PE-CVD) techniques. This article describes: 1- the characterization of modified surfaces using attenuated total reflection-Fourier transform infrared spectroscopy (ATR/FTIR) and contact angle measurements, 2- the results of three in-vitro haemocompatibility assays. Coated surfaces were compared to uncoated materials and various substrates such as polymethylmethacrylate (PMMA), polyethylene (LDPE), polydimethylsiloxane (PDMS) and medical steel (MS). Thrombin generation, blood platelet adhesion and complement convertase activity tests revealed the following classification, from the most to the least heamocompatible surface: Si(rub)/ DLC-Si(rub)/ DLC-Si(waf)/ LDPE/ PDMS/ SiC-Si(waf)/ Si(waf)/ PMMA/ MS. The DLC coating surfaces delayed the clotting time, tended to inhibit the platelet and complement convertase activation, whereas SiC-coated silicon wafer can be considered as thrombogenic. This study has taken into account three events of the blood activation: coagulation, platelet activation and inflammation. The response to those events is an indicator of the in vitro haemocompatibility of the different surfaces and it allows us to select biomaterials for further in vivo blood contacting investigations. PMID- 14562275 TI - Biodegradable synthetic polymers for tissue engineering. AB - This paper reviews biodegradable synthetic polymers focusing on their potential in tissue engineering applications. The major classes of polymers are briefly discussed with regard to synthesis, properties and biodegradability, and known degradation modes and products are indicated based on studies reported in the literature. A vast majority of biodegradable polymers studied belongs to the polyester family, which includes polyglycolides and polylactides. Some disadvantages of these polymers in tissue engineering applications are their poor biocompatibility, release of acidic degradation products, poor processability and loss of mechanical properties very early during degradation. Other degradable polymers such as polyorthoesters, polyanhydrides, polyphosphazenes, and polyurethanes are also discussed and their advantages and disadvantages summarised. With advancements in tissue engineering it has become necessary to develop polymers that meet more demanding requirements. Recent work has focused on developing injectable polymer compositions based on poly (propylene fumarate) and poly (anhydrides) to meet these requirements in orthopaedic tissue engineering. Polyurethanes have received recent attention for development of degradable polymers because of their great potential in tailoring polymer structure to achieve mechanical properties and biodegradability to suit a variety of applications. PMID- 14562276 TI - The precancers: waiting for a classification. PMID- 14562277 TI - Epidermal growth factor receptor expression in follicular dendritic cells: a shared feature of follicular dendritic cell sarcoma and Castleman's disease. AB - The factors regulating the growth of follicular dendritic cell (FDC) sarcoma are currently unknown. Using cDNA microarray analysis, we found that the epidermal growth factor receptor (EGFR) is expressed in FDC sarcoma. We immunohistochemically examined the expression of EGFR in a larger series of FDC sarcomas and in nonneoplastic FDCs. This included 8 cases of FDC sarcoma, 12 cases of hyaline vascular Castleman's disease (CD), 5 cases of human herpesvirus 8 (HHV8)-positive plasma cell CD, 7 cases of HHV8-negative plasma cell CD, 13 cases of reactive lymph nodes, 3 cases of reactive tonsils, 10 cases of follicular lymphoma, 6 cases of nodular mantle cell lymphoma, and 6 cases of angioimmunoblastic T-cell lymphoma. EGFR was expressed in tumor cells in 7 of 8 cases (88%) of FDC sarcoma (strongly in 4 cases and moderately in 3 cases). The single EGFR-negative case had an anaplastic appearance and a more aggressive clinical behavior. EGFR was also expressed by FDC in all types of CD (strongly in 4 cases, moderately in 16 cases, and weakly in 4 cases). Immunostaining results were negative or only weakly positive for EGFR in FDC of reactive lymph nodes and tonsils, and in the FDC networks of follicular lymphoma, mantle cell lymphoma, and angioimmunoblastic lymphoma. The up-regulation of EGFR in FDC of CD was paralleled by an increase in EGFR expression in the surrounding perifollicular fibroblastic reticulum cells suggesting coordinate regulation. These findings identify a differentially expressed growth regulatory receptor common to both FDC sarcoma and CD, identifying a target for possible therapy in unresectable or refractory cases. PMID- 14562278 TI - Possible association between tumor-suppressor gene mutations and hMSH2/hMLH1 inactivation in alveolar soft part sarcoma. AB - Alveolar soft part sarcoma (ASPS) is a rare soft tissue tumor of unknown origin and pathogenesis. We clinicopathologically analyzed 16 cases of ASPS and screened for the genetic alterations of various tumor-suppressor genes and oncogenes, including p53, adenomatous polyposis coli (APC), E-cadherin, and beta-catenin, in 11 cases of ASPS. We also examined the expression of hMSH2/hMLH1 of DNA mismatch repair genes by immunohistochemistry, and promoter hypermethylation of these DNA mismatch repair genes by methylation-specific polymerase chain reaction (MS-PCR) to elucidate any possible association between mutation status of these genes and inactivation of the hMSH2/hMLH1 genes. Furthermore, microsatellite instability (MSI) analysis and loss of heterozygosity (LOH) on chromosome 5q analysis were used for some cases of ASPS where DNA derived from normal tissue was available. The 5-year overall survival rate for all of the patients in this study was 68.6%. The 5-year overall survival rates for patients presenting with localized ASPS and for patients with distant metastases were 83.3% and 47.6%, respectively. The high nuclear grade of tumor cells was a significantly adverse prognostic factor (P = 0.0085). Single-strand conformation polymorphism analysis followed by DNA direct sequencing revealed 4 point mutations of the p53 gene in 3 of 11 cases (27.3%), composed of 3 missense mutations and 1 silent mutation. In addition, 1 case with the E-cadherin missense mutation and 1 case with the APC missense mutations were observed, respectively. None of the cases harbored mutation of exon 3 of the beta catenin gene. Loss of expression of the hMSH2 and hMLH1 genes was observed in 2 (18.2%) and 3 (27.3%) of 11 cases, respectively. All 3 cases with loss of hMLH1 gene expression harbored mutations of the p53 gene. There was a statistically significant correlation between the genetic alteration positive in these tumor suppressor genes and loss of hMLH1 gene expression (P = 0.024). Methylation specific PCR did not reveal hypermethylation of the hMSH2/hMLH1 promoter region in any of the cases examined. Three of 8 (37.5%) ASPS cases showed low MSI, and 2 of these 3 cases showed immunohistochemical lack of expression for either hMSH2 or hMLH1. LOH on 5q was present in 2 of 6 (33.3%) informative cases, and both cases showed LOH on the D5S346 marker, a microsatellite marker near the APC locus. Thus, inactivation of hMSH2/hMLH1 of DNA mismatch repair genes seems to have an important role to play in the mutagenesis of the tumor-suppressor genes in ASPS. PMID- 14562279 TI - The role of p63 and deltaNp63 (p40) protein expression and gene amplification in esophageal carcinogenesis. AB - p63, a member of the p53 gene family, is known to encode functionally antagonistic protein isoforms. Although transactivating protein isoforms display p53-like functions, deltaNp63 isoforms act toward p53 in a dominant negative way. Using immunohistochemistry, we examined the expression of pan-p63 and deltaNp63 in 50 esophageal squamous cell carcinomas (SCCs) as well as in squamous low-grade intraepithelial neoplasias (S-LGINs; n = 4) and high-grade intraepithelial neoplasias (S-HGINs; n = 18). Additionally, 50 esophageal adenocarcinomas (ADCs) that arose in Barrett's esophagus (BE) as well as adjacent specialized metaplastic epithelium (SE; n = 41), low-grade intraepithelial neoplasias (B LGINs; n = 27), and high-grade intraepithelial neoplasias (B-HGINs; n = 21) in BE were investigated. Furthermore, p63 gene amplification was determined by fluorescent differential polymerase chain reaction in a subset of 10 SCCs and 10 ADCs. Whereas in normal esophageal epithelium, expression of pan-p63 is invariably restricted to the basal cell layer, in 100% of S-LGINs, 94.4% of S HGINs, and 88.0% of SCCs, expression of p63 was found in >75% of the cells. Concerning BE, only in a small subset of SEs (7.3%), B-LGINs (14.8%), B-HGINs (14.3%) and ADCs (16.0%) was a weak p63 protein expression (<10% positive cells) detectable, whereas the rest of the samples were completely negative. Expression of deltaNp63 was identical to expression of pan-p63 in the vast majority of samples. p63 gene amplification was found in 2 of 10 (20.0%) investigated SCCs and in 1 of 10 (10.0%) ADCs. In summary, strong expression of p63, especially its deltaNp63 isoforms, is a frequent finding in esophageal precancerous and cancerous squamous lesions, whereas this is not the case in carcinogenesis of BE. p63 gene amplification is an infrequent finding in esophageal SCCs and ADCs and does not correlate with protein overexpression. PMID- 14562280 TI - Reduced expression of syndecan-1 correlates with histologic dedifferentiation, lymph node metastasis, and poor prognosis in intrahepatic cholangiocarcinoma. AB - Syndecan-1, a cell-surface transmembrane heparan sulfate proteoglycan, has been reported to correlate with the biologic behavior of malignant tumors in various organs. We examined the correlation between the expression of syndecan-1 at the protein and mRNA levels and clinicopathologic features of 37 intrahepatic cholangiocarcinomas (ICCs). Noncancerous bile duct epithelial cells showed basolateral membranous expression of syndecan-1, whereas ICC cells showed membranous and also diffuse cytoplasmic expression. In situ hybridization demonstrated a distribution of syndecan-1 mRNA similar to that of the protein in carcinoma tissue, suggesting that syndecan-1 expression in ICC is regulated at the transcriptional level. Reduction of syndecan-1 expression in carcinoma was associated with poor histological differentiation (P <0.01): syndecan-1 expression was intense and extensive in well-differentiated (10 cases) and largely negative or weakly positive in poorly differentiated (13 cases) adenocarcinoma, and its expression in moderately differentiated tumors (14 cases) was intermediate. Patients with ICCs demonstrating negative or weak expression of syndecan-1 frequently had lymph node metastases and had a rather poor prognosis after surgical resection compared with those whose tumors demonstrated moderate or strong expression (P <0.05). However, syndecan-1 expression was not correlated with tumor size, stromal desmoplasia, gross classification, vascular invasion, or perineural invasion. We conclude that expression of syndecan-1 could correlate with some aspects of the biologic behaviors of ICCs and may be a useful prognostic marker. PMID- 14562282 TI - Chromosome 22q alterations and expression of the NF2 gene product, merlin, in gastrointestinal stromal tumors. AB - Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the digestive tract. A characteristic genetic alteration in GISTs is constitutive activation of the c-kit proto-oncogene, but alterations in chromosomes 14 and 22 may also play a role in the molecular pathogenesis. In this study, 42 GISTs were analyzed for loss of heterozygosity (LOH) on the long arm of chromosome 22 (22q). Overall, 69% of the tumors studied showed LOH with at least 1 of the 22q markers. Allele losses were compared with tumor mitotic activity, the most commonly used prognostic marker for this tumor. Interestingly, allele deletion at 22q was significantly more frequent in tumors with high mitotic activity (>/= l2 mitoses/10 high-power fields [HPF]) than in tumors with low mitotic activity (< 2 mitoses/HPF)-88% versus 56% (P < 0.01). A total of 26% (11 of 42) of all tumors demonstrated loss of all 22q sites analyzed, consistent with the loss of 1 copy of the entire long arm. Such tumors carried a 4.6-fold (95% confidence interval, 0.5 to 49.8) risk for recurrence compared with tumors with no LOH. LOH was frequently detected at the neurofibromatosis 2 (NF2) tumor-suppressor gene locus at 22q12. Sequencing of the NF2 gene from 5 GISTs did not reveal mutations, however. Furthermore, 16 of 19 tumors (84%) analyzed by immunohistochemistry were positive for the NF2 gene product, merlin. The findings suggest that allelic losses at 22q are associated with high mitotic activity and recurring disease, and that alterations in the NF2 gene are unlikely to participate in the pathogenesis of GIST. PMID- 14562281 TI - Comparative genomic hybridization analysis of hepatoblastoma reveals high frequency of X-chromosome gains and similarities between epithelial and stromal components. AB - Hepatoblastoma (HB) is the most common liver tumor in childhood and differs in its environmental risk factors and genetic background from hepatocellular carcinoma. HB is associated with inherited conditions such as familial adenomatous polyposis and Beckwith-Wiedemann syndrome, suggesting the importance of genetic abnormalities in the pathogenesis and progression of this disease. It has a very polymorphous morphology. A diverse range of cytogenetic alterations has been reported to date, the most frequent being trisomy 2 and trisomy 20. Thirty-five HB specimens from 31 patients (22 purely epithelial, 4 purely mesenchymal, 9 mixed) were examined by comparative genomic hybridization (CGH), a technique that enables us to screen the entire tumor genome for genetic losses and gains. Our aims were as follows: (1) to characterize chromosome abnormalities that appear in this tumor and (2) to identify possible differences between different histologic subtypes of HB. We found significant gains of genetic material, with very little difference in the number and type of alterations between the different histologic components of HB. The most frequent alterations were gains of Xp (15 cases, 43%) and Xq (21 cases, 60%). This finding was also confirmed by fluorescent in situ hybridization performed on nuclei extracted from 6 specimens. Other common alterations were 1p-, 2q+, 2q-, 4q-, and 4q+. We found no difference between different histologic subtypes, a finding that may be in agreement with the hypothesis of a common clonal origin for the different components. An hitherto-unreported high frequency of X chromosome gains may support the assumption that X-linked genes are involved in the development of this neoplasm. PMID- 14562283 TI - Overexpression of p53 is correlated with stromal invasion in extramammary Paget's disease of the vulva. AB - Molecular alterations that are associated with clinicopathological features of extramammary Paget's disease of the vulva (PDV) are poorly understood. Consequently, we have investigated whether a correlation exists between overexpression of p53 protein and various clinicopathologic features of PDV. Our study group comprises 10 primary noninvasive PDVs, 3 primary PDVs with minimal invasion, and 1 primary PDV with frank invasion. Recurrence in the form of noninvasive PDV was seen in 4 patients with previously noninvasive PDVs and in 1 patient who previously had PDV with minimal invasion. Metastases to the inguinal lymph nodes were associated with the 1 PDV with frank invasion and 1 of the PDVs with minimal invasion. An immunohistochemical study of p53 expression was performed on paraffin-embedded tissue. Negative p53 immunostaining was seen in all of the primary noninvasive PDVs as well as their recurrences. Positive p53 immunostaining was observed in the invasive as well as the intraepidermal components of all of the primary PDVs with invasion, the metastatic tumors in the inguinal lymph nodes, and the recurrent PDV associated with prior invasion, indicating a possible role of p53 in the progression of PDV. To our knowledge, our observation of p53 overexpression in the intraepidermal component of PDVs associated with invasion is the first to be reported in the literature. This observation may prove helpful in identifying stromal invasion in small biopsies. We also report for the first time an association between high nuclear grade in PDV and the propensity for inguinal lymph node metastasis. PMID- 14562284 TI - Human papillomavirus infection in actinic keratosis and bowen's disease: comparative study with expression of cell-cycle regulatory proteins p21(Waf1/Cip1), p53, PCNA, Ki-67, and Bcl-2 in positive and negative lesions. AB - We examined the presence of human papillomavirus (HPV) DNA in tissues of premalignant skin lesions, i.e., actinic keratosis (n = 13) and Bowen's disease (n = 62), taken from 69 Japanese immunocompetent and renal transplant recipient patients. Detection and typing of HPV DNA were performed using polymerase chain reaction (PCR) and sequence analysis or restriction fragment length polymorphism (RFLP) analysis, respectively. The positivity rates of HPV DNA in tissues of actinic keratosis and Bowen's disease were 77% and 65%, respectively. Twenty seven HPV types were detected in 50 (67%) premalignant skin lesions, in which Z95963 (accession no. in the EMBL Databank), Z95968, AJ010823, and AJ000151 have been described as partial sequences of unknown HPV types. Furthermore, 2 unknown types, HPVX1 and HPVX2, were found in specimens of actinic keratosis. Sequence analysis showed that HPVX1 is related to HPV-37 (86.1% sequence homology) and that HPVX2 is related to HPV-38 (79.7%). These results indicate that various mucosal and epidermodysplasia verruciformis-related HPV types are associated with the pathogenesis of actinic keratosis and Bowen's disease. In addition, 24 specimens of HPV-positive or HPV-negative premalignant skin lesions were examined immunohistochemically for proliferating cells to determine biological differences between HPV-positive and HPV-negative lesions. Immunohistochemistry for p21(Waf1/Cip1), p53, proliferating cell nuclear antigen (PCNA), Ki-67, and Bcl-2 revealed that there was no significant difference in the cell proliferation activity between HPV-positive and HPV-negative lesions, suggesting that HPV infection alone does not induce cell proliferation in those lesions. PMID- 14562285 TI - Subvisual changes in chromatin organization state are detected by karyometry in the histologically normal urothelium in patients with synchronous papillary carcinoma. AB - This study analyzed the chromatin organization state in histologically normal urothelium in patients with synchronous papillary carcinoma using digital texture analysis. The quantitative evaluation was carried out on hematoxylin and eosin stained sections from 17 cases of urothelial papillary carcinoma in which a simultaneous biopsy specimen featuring histologically normal urothelium was available. Five bladder biopsy specimens of histologically normal urothelium from patients with prostate pathology in whom cystoscopy revealed a normal bladder mucosa were also analyzed. Karyometry showed that the 17 cases of papillary carcinoma, morphologically classified according to the 1973 World Health Organization scheme, belonged to a continuous spectrum or trend curve spanning grade 1 to grade 3. An abnormal pattern and distribution of the nuclear chromatin was seen in the normal-looking urothelium from the 17 bladders with papillary lesions. When this population was plotted along the trend curve, it occupied an intermediate position between the normal samples and samples from grade 1 carcinoma. When the nuclei were considered individually, the changes were detected only in a subpopulation of nuclei with chromatin alteration pointing toward that seen in grade 1 cases, even though distinct from them. In conclusion, karyometry can detect an abnormal chromatin pattern and distribution in the normal-looking urothelium adjacent to papillary carcinoma. Such alterations correspond to the so-called "malignancy-associated change." PMID- 14562286 TI - Molecular and immunohistochemical analysis of intraductal papillary neoplasms of the biliary tract. AB - Intraductal papillary neoplasms (IPNs) of the biliary tract are uncommon lesions that may be solitary or may spread extensively along the biliary tree. Some biliary IPNs are histologically and radiologically similar to intraductal papillary mucinous tumors (IPMNs) of the pancreas and present a risk for progression to invasive cholangiocarcinoma. Unlike pancreatic IPMNs, little is known about their molecular pathogenesis. We studied 14 biliary IPNs (including 5 cases with associated invasive cholangiocarcinoma) for genetic alterations in the APC/beta-catenin pathway, K-ras oncogene mutations, p53/chromosome 17p alterations, and Dpc4/18q alterations. Immunohistochemistry was performed for beta-catenin, p53, and Dpc4, and microdissected tissue was analyzed using direct DNA sequencing for exon 1 of K-ras and exon 3 of beta-catenin and allelic loss assays on chromosomes 5q, 17p, and 18q. Activating mutations in codon 12 of the K ras oncogene were present in 4 of 14 (29%) biliary IPNs. Of these 4 cases, 2 patients had associated invasive cholangiocarcinoma, and identical K-ras mutations were present in both the intraductal and invasive components. Allelic loss on chromosome 18q was present in 4 of 13 informative cases (31%); however, no loss of normal Dpc4 expression was detected by immunohistochemistry. Nuclear accumulation of beta-catenin protein was demonstrated in 3 of 12 cases (25%); however, there were no beta-catenin gene mutations, and allelic loss on 5q was present in only 1 of 10 informative cases (10%). Both immunohistochemistry for p53 and 17p allelic loss assays were negative. Biliary IPNs therefore demonstrate a K-ras gene mutation frequency that is lower than that previously reported for pancreatic IPMNs, but similar to that reported for hepatic cholangiocarcinomas. The presence of K-ras mutations in 2 purely intraductal neoplasms, and identical K-ras mutations in 2 cases with both intraductal and invasive components, suggests that these mutations arise early in tumorigenesis. Finally, the frequency of allelic loss on 18q suggests that a locus on 18q is involved in the molecular pathogenesis of biliary IPNs, but this locus is not DPC4. PMID- 14562287 TI - Should each core with prostate cancer be assigned a separate gleason score? AB - If multiple biopsy cores contain prostate cancer with differing Gleason scores, should an overall Gleason score be assigned, or should each core be graded separately? We obtained data on 127 men with prostate cancer on needle biopsy who underwent subsequent radical prostatectomy at our institution. We compared the Gleason scores found on needle biopsy with the grade and stage (organ-confined, extra-prostatic extension, positive seminal vesicles or lymph nodes) at radical prostatectomy. On biopsy, 40 men had a pure Gleason score of 4 + 3 = 7, 25 men had a Gleason score of 4 + 3 = 7 with a Gleason score of 3 + 3 = 6 on a separate core of the biopsy specimen, 27 men had a pure Gleason score of 4 + 4 = 8, and 35 men had a Gleason score of 4 + 4 = 8 with separate cores containing Gleason pattern grade 3. A Gleason score of 4 + 4 = 8 with pattern grade 3 in other cores had a more advanced stage than a pure Gleason score of 4 + 3 = 7 (P = 0.008). There was no clear pattern analyzing pathological stage of men with a pure Gleason score of 4 + 3 = 7 in comparison with those with Gleason scores of 4 + 3 = 7 and 3 + 3 = 6 in other cores. The group with a Gleason score of 4 + 4 = 8 and Gleason pattern grade 3 on other cores had a higher overall grade on radical prostatectomy than the group with a pure Gleason score of 4 + 3 = 7 (P = 0.001). If one had assigned an overall Gleason score, then a biopsy with Gleason score 4 + 4 = 8 on 1 or more cores and some pattern grade 3 in other cores, would be designated as a Gleason score of 4 + 3 = 7. Based on our findings, patients with a Gleason score of 4 + 4 = 8 on one or more cores with pattern grade 3 in other cores should be given a final Gleason score of 4 + 4 = 8 instead of 4 + 3 = 7, because these patients are more likely to have higher stage and grade on radical prostatectomy, comparable to a pure Gleason score of 4 + 4 = 8. Each core should be assigned a separate Gleason score, especially in cases with high Gleason score cancer on at least 1 core. PMID- 14562288 TI - Identification of chromosomal copy number changes associated with transformation of follicular lymphoma to diffuse large B-cell lymphoma. AB - The histological transformation from a follicular lymphoma (FL) to a diffuse large B-cell lymphoma (DLBL) occurs in 22% to 30% of all cases of FL. The aim of this study was to identify specific chromosomal gains/losses associated with transformation of FL to DLBL, in addition to the well-known mechanisms like p53 mutation and protein expression and c-myc translocation and up-regulation. This is the first study to meet 2 important conditions for such a comparison. First, we demonstrate that the FL and the DLBL were clonally related, based on identical immunoglobulin gene rearrangements in 5 of the 6 cases. Second, we used laser microdissection microscopy to isolate only the neoplastic cells from the initial FL samples. The results indicate that no single chromosomal abnormality seems to be responsible for the transformation of FL to DLBL. P53 protein overexpression was found in 4 and c-myc translocation in 3 of the 6 transformed DLBLs, but not in the initial FL samples. Additional chromosomal abnormalities were detected by comparative genomic hybridization in all 6 cases when the DLBL was compared with the FL. In the 5 cases with transformation of grade 1 or 2 FL to DLBL, gains at chromosomes 7 (5 of 5 cases), 10p1 (3 of 5 cases), 12 (3 of 5 cases), and 20p13 (2 of 5 cases) and loss at 9q (4 of 5 cases) were the most frequently found abnormalities. A gain on chromosome 7p, in combination with a loss on 9q, was found in 4 of the 5 DLBL that transformed from FL grade 1 or 2. PMID- 14562289 TI - The role of replicative senescence in chronic allograft nephropathy. AB - Strong evidence suggests that replicative senescence is involved in vivo because senescent cells have been detected in human tissues associated with physiological and pathological aging processes. Chronic allograft nephropathy (CAN) appears to be a major determinant of long-term survival in kidney transplantation. Several mechanisms are potentially involved; the aim of this study was to assess the impact of replicative senescence in CAN. Replicative senescent cells were detected on renal tissue cryosection using expression of a specific marker, senescence-associated beta-galactosidase (SA-beta-Gal) at pH 6. A total of 80 frozen renal samples (67 cases of CAN and 13 controls) were studied. To validate this marker, we measured in situ telomere length in cells expressing or not expressing SA-beta-Gal using a validated quantitative fluorescence in situ hybridization technique. The presence of senescent cells was correlated with clinicopathologic data. Telomere length was significantly lower in cells expressing SA-beta-Gal than in cells that did not. Replicative senescence was present in 45 out of 67 (67%) biopsy specimens and was significantly associated with the severity of CAN. No correlation with the notion of a previous episode of acute tubular necrosis, acute rejection, extrarenal epuration, duration of cold ischemia, and the delay between transplantation and biopsy was observed. However, the age of the donor, but not that of the recipient, was correlated with the occurrence of senescent cells. These results suggest that replicative senescence is a mechanism that might be involved in the development of CAN. The age of the donor appears to be the major determinant factor in replicative senescence. PMID- 14562290 TI - Pseudomonas pneumonia in infants: an autopsy study. AB - Pseudomonas pneumonia is an uncommon but serious infection in infants, occurring mainly in infants of low birth weight. In this retrospective clinicopathologic correlation study, we reviewed the clinical records and analyzed postmortem lung pathology in 8 infants with pneumonia due to P. aeruginosa. From the histopathology, 2 different pneumonic patterns emerged: a distinctive paucicellular coagulative confluent bronchopneumonia with perivascular bacillary infiltration (7 cases) and a more usual cellular pneumonia without evidence of perivascular organisms (1 case). Clinically, infants with the first type could be considered immunocompromised and had a precipitous course characterized by signs of sepsis, whereas the infant with the second type (who likely had a more normal immune system) had a relatively protracted course with respiratory failure. We conclude that (1) the pattern of pneumonic inflammation correlates with the immune state of infants, similar to what has been reported in adults; (2) among immunocompromised infants, histopathologic signs of bacteremia are prevalent; and (3) the clinical signs do not correlate with the severity of the pathology at autopsy and may reflect sepsis rather than pneumonia. We speculate that the histopathology in this population reflects the virulence of the organism, as well as the immune status of the host. PMID- 14562291 TI - Uterine serous carcinoma and endometrial intraepithelial carcinoma arising in endometrial polyps: report of 5 cases, including 2 associated with tamoxifen therapy. AB - Uterine serous carcinoma (USC) is the prototype of type II endometrial cancer. Endometrial intraepithelial carcinoma (EIC) is the precursor lesion of USC, Rarely, USC and EIC may arise within and be largely confined to otherwise benign endometrial polyps. This report describes 5 such cases. The patients ranged in age from 67 to 89 years, with a mean age of 75 years. In 2 of the cases there was a history of tamoxifen therapy. In 2 cases USC or EIC was confined to the endometrial polyp, and in 3 cases there was focal involvement of nonpolypoid endometrium. In 1 case there was a single small focus of extrauterine tumor within an ovarian vascular channel. In 2 cases the invasive tumor within the polyp also contained areas of endometrioid adenocarcinoma, and in 2 cases there was a component of clear cell carcinoma. In all cases USC and EIC were strongly reactive for p53 and showed a high proliferation index with MIB1. Two cases were negative with estrogen receptor, and 3 cases exhibited positive staining. The cases reported herein show that USC and EIC may rarely arise in benign endometrial polyps and that extrauterine involvement may be present without myometrial infiltration. Because 2 of the patients had been taking tamoxifen, this raises the possibility of an association between tamoxifen and the development of USC and EIC in the endometrial polyps that are characteristic of this medication. PMID- 14562292 TI - Enterococcus gastritis. AB - Helicobacter pylori infection is the most common cause of gastritis with its associated sequelae. Gastritis secondary to other bacteria is rare. This report describes Enterococcus-associated gastritis in a 59-year-old diabetic man. Nine months after receiving treatment for H. pylori-associated gastritis, he underwent endoscopy to confirm H. pylori eradication and to evaluate the status of previously seen ulcers. Mucosal biopsy specimens revealed severe active but focal gastritis adjacent to gram-positive coccobacilli in short to long chains with no H. pylori. Culture grew an Enterococcus similar to E. hirae and E. durans. No treatment was given, and endoscopy done 2 months later showed complete resolution of the gastritis and absence of H. pylori or enterococci. Our patient's gastritis represents a previously undescribed manifestation of Enterococcus infection. It is possible that the presence of NSAID gastric mucosal injury and diabetes predisposed this individual to the development of transient Enterococcus gastritis. PMID- 14562293 TI - Genome profiles of bilateral dysgerminomas, a unilateral gonadoblastoma, and a metastasis from a 46, XY phenotypic female. AB - We present a case report of a 16-year-old, phenotypic female with bilateral dysgerminomas, a unilateral gonadoblastoma, and a peritoneal metastasis. The patient's constitutional karyotype was 46,XY. The chromosomal copy number, examined by the comparative genomic hybridization technique, showed 3 gains in the dysgerminoma of the right ovary, 6 gains in the dysgerminoma of the left ovary, and 2 gains and 1 loss in the gonadoblastoma of the left ovary. The metastasis showed 5 gains of which 4 were also observed in the dysgerminoma of the left ovary. The DNA ploidy classifications of the gonadoblastoma and the dysgerminoma in the right ovary were tetraploid, whereas the dysgerminoma in the left ovary and the metastasis were aneuploid. We therefore propose that the metastasis most probably developed from the dysgerminoma of the left ovary. PMID- 14562294 TI - Epstein-Barr virus-associated intravascular large T-cell lymphoma presenting as acute renal failure in a patient with acquired immune deficiency syndrome. AB - Intravascular lymphoma (IVL) is a rare neoplasm, recently included as a specific entity in the World Health Organization classification of lymphoid tumors. Most cases are of B-cell lineage; however, rare cases of T-cell phenotype have been reported. We report a human immunodeficiency virus (HIV)-positive patient who died of acute renal failure in whom IVL was identified at autopsy, predominantly involving the renal interstitial vessels. Immunohistochemical stains revealed a T cell phenotype, which was confirmed by T-cell receptor gamma gene rearrangement studies. The lymphoma cells showed nuclear Epstein-Barr virus (EBV)-encoded RNA transcripts by in situ hybridization, suggesting that EBV might be of etiologic importance in this tumor. The predominant involvement of kidney is unusual. With effective therapy, morbidity and mortality of HIV-1 infection has been substantially reduced, and survival times have been prolonged. However, the relative risk of secondary neoplasms, especially non-Hodgkin's lymphoma (NHL), has increased. Consequently, we conclude that unique types of NHL, such as this case of IVL, may be encountered more frequently in this patient population, and that NHL should be added to the list of differential diagnostic considerations in HIV-1-positive patients who develop acute renal failure. PMID- 14562295 TI - Fibroblastic reticular cell tumor of the spleen: report of a case and review of the entity. AB - Fibroblastic reticulum cells (FBRCs) are stromal support cells located in the parafollicular area and deep cortex of lymph nodes and in the extrafollicular areas of the spleen and tonsils. We report a case of malignant FBRC tumor of the spleen occurring in a 61-year-old woman. Two years after splenectomy, multiple hepatic lesions were found, which were resected. Histologically, the tumor showed similar morphological features in the spleen as in the liver metastases. There was a whorled pattern of oval and spindle cells in a collagenized background admixed with an inflammatory cell infiltrate composed of lymphocytes and plasma cells. The tumor cells were positive for common muscle actin, smooth muscle actin, and focally for CD68. In situ hybridization for Epstein Barr virus was negative. To the best of our knowledge, this is the first report of malignant FBRC tumor arising in the spleen. The differential diagnosis of splenic tumors with inflammatory pseudotumor-like features is discussed. PMID- 14562296 TI - Armed forces institute of pathology update. PMID- 14562297 TI - Growth index is independent of microvessel density in non-small cell lung carcinomas. PMID- 14562298 TI - Renal epithelial neoplasms: the diagnostic implications of electron microscopy study in 55 cases. PMID- 14562302 TI - Structure and reactivity of Ziegler-Natta catalyst intermediates. AB - This paper provides a survey of our recent research on chemistry that occurs among components of Ziegler-Natta catalysts. Since the amount of information regarding this topic is enormous we have focused on alkoxo compounds, which are attracting more and more attention in polymer industry. In this concept article we discuss recent studies on the inherently very complex catalyst system that involve isolation of various solid polynuclear metal-containing species, determination of their structures by X-ray crystallography, and relating their structures to the activity in polymerization. The presented research may be useful in predicting trends for the development of new single-site catalysts as well as new co-catalysts. PMID- 14562303 TI - Formation of quaternary stereocenters by copper-catalyzed Michael reactions with L-valine amides as auxiliaries. AB - Extensive investigations of chiral auxiliaries and active metals for Michael addition of 1,3-dicarbonyl compounds with vinyl ketones are summarized. Our efforts result in a widely applicable auxiliary-mediated, copper(II) acetate catalyzed procedure. For these purposes, L-valine diethylamide is an optimal chiral auxiliary giving quaternary stereocenters with up to 99 % ee at ambient temperature. No inert or anhydrous conditions are required, the solvent is simply acetone, and the auxiliary can be recovered almost quantitatively after workup. PMID- 14562304 TI - "Janus" supermolecular liquid crystals--giant molecules with hemispherical architectures. AB - Liquid crystals represent a unique class of self-organising systems, which although found in many day-to-day practical material applications, such as displays, are also intimately entwined with living processes. They have the potential, just like living systems, to provide us with a unique vehicle for the development of self-ordering nano- and mesoscopic-engineered materials with specific functional properties. In this article we describe a new concept for the design of self-assembling functional liquid crystals as segmented or "Janus" liquid-crystalline supermolecular materials in the form of structures that contain two different types of mesogenic units, which favour different types of mesophase structure, grafted onto the same star-shaped scaffold to create supermolecules that contain different hemispheres. The materials exhibit chiral nematic and chiral smectic C phases. PMID- 14562305 TI - Photochromism of diarylethenes linked by hydrogen bonds in the single-crystalline phase. AB - Photochromic diarylethenes, which bear carboxyl groups at the ortho, meta, or para positions of both terminal phenyl groups, have been synthesized. The diarylethenes adopt linear chain structures as a result of hydrogen bonding in the crystalline phase, and the crystals exhibit photochromic properties. The absorption maximum of the photogenerated closed-ring isomer of the para substituted derivative shows an 80 nm bathochromic shift in comparison with that of the ortho-substituted derivative. The maximum of the closed-ring isomer of the meta-substituted derivative is located in between those of the para- and ortho substituted derivatives. The shifts can be attributed to the differences in conformation among the derivatives, arising from the restrictions imposed by the hydrogen-bonded chains. PMID- 14562306 TI - Stereoselective aldol additions catalyzed by dihydroxyacetone phosphate-dependent aldolases in emulsion systems: preparation and structural characterization of linear and cyclic iminopolyols from aminoaldehydes. AB - The potential of dihydroxyacetone phosphate (DHAP)-dependent aldolases to catalyze stereoselective aldol additions is, in many instances, limited by the solubility of the acceptor aldehyde in aqueous/co-solvent mixtures. Herein, we demonstrate the efficiency of emulsion systems as reaction media for the class I fructose-1,6-bisphosphate aldolase (RAMA) and class II recombinant rhamnulose-1 phosphate aldolase from E. coli (RhuA)-catalyzed aldol addition between DHAP and N-benzyloxycarbonyl (N-Cbz) aminoaldehydes. The use of emulsions improved the RAMA-catalyzed aldol conversions by three to tenfold relative to those in conventional DMF/water mixtures. RhuA was more reactive than RAMA towards the N Cbz aminoaldehydes regardless of the reaction medium. With (S)- or (R)-Cbz alaninal, RAMA exhibited preference for the R enantiomer, while RhuA had no enantiomeric discrimination. The linear N-Cbz aminopolyols thus obtained were submitted to catalytic intramolecular reductive amination to afford the corresponding iminocyclitols. This reaction was diastereoselective in all cases examined; the face selectivity was controlled by the stereochemistry of the newly formed hydroxyl group originating from the aldehyde. Characterization of the resulting iminocyclitols allowed the assessment of the diastereoselectivity of the enzymatic aldol reactions with respect to the N-protected aminoaldehyde. RAMA formed single diastereoisomers from N-Cbz-glycinal and from both enantiomers of N Cbz-alaninal, while 14 % of the epimeric product was observed from N-Cbz-3 aminopropanal. Diastereoselectivity from RhuA was lower than that observed from RAMA. Interestingly, a single diastereoisomer was formed from (S)-Cbz-alaninal, whereas only a 34 % diastereomeric excess was observed from its enantiomer (i.e., (R)-Cbz-alaninal). PMID- 14562307 TI - The effect of hydrogen bonding on allylic alkylation and isomerization reactions in ionic liquids. AB - Neutral allylic alkylation reactions, in which a base is generated in situ and which hence require no external bases, can significantly be retarded when carried out in the ionic liquid 1-butyl-3-methylimidazolium tetrafluoroborate ([bmim][BF(4)]). Evidence suggests that the base or base precursor enters into hydrogen bonding with the imidazolium cation and is thus made less readily available for deprotonation of pre-nucleophiles. However, the reaction proceeds well in the presence of stronger bases that are capable of deprotonation. Whilst the phenomenon of hydrogen bonding in ionic liquids can be detrimental to reactions such as allylic alkylation, it can be exploited to suppress unwanted allylic isomerization. PMID- 14562308 TI - Efficient transport of saccharides through a liquid membrane mediated by a cyclodextrin dimer. AB - A new efficient system for transporting saccharides through a liquid membrane has been constructed. The transport rates of saccharides were accelerated greatly by the cyclodextrin dimer 2; by contrast, the corresponding cyclodextrin monomer 1 was not effective at mediating saccharide transport. The transport rate of D ribose through a chloroform liquid membrane was 17 times faster when the cyclodextrin dimer 2 was used as the transporter than when the cyclodextrin monomer 1 was used. Similarly the transport rate of methyl D-galactopyranoside was 16 times faster by 2 than by 1. PMID- 14562309 TI - The hetero-Diels-Alder addition of sulfur dioxide: the pseudo-chair conformation of a 4,5-dialkylsultine. AB - Even unsubstituted butadiene adds to sulfur dioxide in the hetero-Diels-Alder mode more rapidly than in the chelotropic mode. The sultine can be observed in equilibrium with the diene and the sulfur dioxide only at low temperature and in the presence of CF(3)COOH. Crystals of 4,5-dialkyl-sultine resulting from the SO(2) addition to 1,2-dimethylidenecyclohexane have been obtained at -100 degrees C and analyzed by X-ray diffraction. Quantum chemical calculations have shown that hyperconjugative interactions within the sulfinyl moiety are responsible for the anomeric effects observed in sultines that prefer pseudo-chair conformations with pseudo-axial Sdbond;O bonds. PMID- 14562310 TI - Structure and reactivity of homoleptic samarium(II) and thulium(II) phospholyl complexes. AB - Potassium 2,5-di-tert-butyl-3,4-dimethylphospholide K(dtp) (9) was synthesised in 45 % yield from commercially available starting materials by using zirconacyclopentadiene chemistry. Reaction of the K salt of this bulky anion and of the previously described potassium 2,5-bis(trimethylsilyl)-3,4 dimethylphospholide K(dsp) (8) with SmI(2) in diethyl ether afforded the homoleptic samarium(II) complexes 7 and 6, respectively, whose solid-state structures, [[Sm(dtp)(2)](2)] (7 a) and [[Sm(dsp)(2)](2)] (6 a), are dimeric owing to coordination of the phosphorus lone pairs to samarium, as shown by X-ray crystallography. Reaction of 8 with TmI(2) in diethyl ether afforded [Tm(dsp)(2)(Et(2)O)], which could not be desolvated without decomposition. In contrast, the coordinated ether group of the solvate [Tm(dtp)(2)(Et(2)O)], obtained from 9 and TmI(2), could easily be removed by evaporation of the solvent and extraction with pentane at room temperature, and the monomer [Tm(dtp)(2)] (5) could be isolated and was characterised by X-ray crystallography. Presumably, steric crowding in 5 is too high for dimerisation to occur. Compound 5, the first Tm(II) homoleptic sandwich complex, is remarkably stable at room temperature in solution and did not noticeably react with nitrogen, in sharp contrast with other thulium(II) species. As expected, 5, 6 and 7 all reacted with azobenzene to give the trivalent complexes [Tm(dtp)(2)(N(2)Ph(2))] (13), [Sm(dsp)(2)(N(2)Ph(2))], (14) and [Sm(dtp)(2)(N(2)Ph(2))] (15), respectively; 13 and 14 were characterised by X-ray crystallography. Complex 5 immediately reacted with triphenylphosphane sulfide at room temperature to give [[Tm(dtp)(2)](2)(mu-S)] (16), which was characterised by X-ray crystallography, whereas samarium(II) complexes 6 and 7 did not noticeably react with Ph(3)PS over 24 h under the same conditions. PMID- 14562312 TI - Characterisation of four new two-dimensional lithium beryllofluoro-layered compounds. AB - Four new amine-templated materials, containing two-dimensional lithium beryllofluoride sheets of the stoichiometry [LiBeF(4)](-), have been synthesised under hydrothermal and ambient pressure conditions. [LiBeF(4)][C(6)H(4)(CH(3))CH(2)NH(3)] (1), [LiBeF(4)][C(6)H(4)CH(2)NH(3)Cl] (2), [LiBeF(4)](2)[NH(3)CH(2)CH(2)CH(2)NH(3)] (3), and [LiBeF(4)][C(6)H(5)CH(2)CH(2)CH(2)NH(3)] (4) all contain well-separated anionic sheets containing two different topologies with the 'inter-layer' regions comprising of organoamine templating species. Use of the different organoamine templating agents results in compounds possessing very different relative arrangements of the lithium beryllofluoride sheets. The materials crystallise in P-centred orthorhombic and monoclinic cells; for 1 (templating agent: 3 methylbenzylamine) Pca2(1); for 2 (4-chlorobenzylamine) Pbca; for 3 (1,3 diamminopropane) Pccn, and for 4 (3-phenyl-1-propylamine) P2(1)/c. Hydrogen bonding exists between ions situated on the protonated amine groups on the templating species and electronegative fluoride ions, on MF(4) tetrahedra (where M=Li and Be). PMID- 14562311 TI - A new family of insulin-mimetic vanadium complexes derived from 5 carboalkoxypicolinates. AB - The reaction of 5-carboalkoxypicolinic acid (5 ROpicH, R=Me, Et, iPr, sBu; 1 a-d) with vanadyl sulfate yielded the complexes [VO(H(2)O)(5 ROpic)(2)], 2 a-d, with H(2)O and one of the picolinato ligands in the equatorial positions, and the second picolinate occupying equatorial (N) and axial (O) positions. Reaction of 1 a with [NH(4)][VO(3)] yielded [NH(4)][VO(2)(5 MeOpic)(2)], [NH(4)]-3, in which the N functions of the picolinates are trans to the doubly bonded, cis-positioned oxo groups. Complexes 1 a.H(2)O, 1 b, 1 c, 2 a.3.5 H(2)O and [NH(4)]-3.4 H(2)O have been structurally characterised. A detailed pH-potentiometric solution speciation analysis of the system VO(2+)-1 a revealed a dominance of VO(5 OMepic)(2) between pH 2 and 6, with the same coordination pattern, evidenced by EPR spectroscopy, as in the crystalline solid state. In ternary systems containing physiological concentrations of the low molecular mass biogenic binders (B) lactate, oxalate, citrate or phosphate, ternary species of general composition VO(5 MeOpic)B dominate at physiological pH, with citrate being the most effective competitor for picolinate. All of the complexes trigger glucose uptake and degradation by simian virus modified mice fibroblasts at non-toxic concentrations (<100 microM), with 2 a, [VO(2)(pic)(2)](-) and [VO(2)(dipic)](-) being at least as effective as insulin. Vanadium uptake by the cells is most effective in the case of 2 a. 2 a also effectively inhibits free fatty acid release by rat adipocytes treated with epinephrine, thus mimicking the inhibition of lipolysis by insulin. PMID- 14562313 TI - Unexpected reaction pathways in the reaction of alkoxyalkynylchromium(0) carbenes with aromatic dinucleophiles. AB - Thermal- or SiO(2)-induced reactions of the Michael adducts of 1,2-aromatic dinucleophiles and alkynylchromium(0) carbene complexes, compounds 7-10, form different products in good yields depending on the nature of the aromatic dinucleophile used. Thus, 1,2-diaminobenzene derivatives 7 and 8 rearrange to pentacarbonylchromium(0) isocyanide complexes 11, 12, 14, and 15 in a process that occurs through bicyclic intermediates 24. Adducts 9 derived from o aminophenol give 2,3-dihydro-1,5-benzoxazepine derivatives 17 by intramolecular 1,2-addition, followed by protonation at the chromium center and reductive elimination. In contrast, base-promoted addition of the phenolic hydroxy group in compound 9 a affords 3-ethoxy-5-phenyl-5,6-dihydro-2H-1,6-benzoxazocin-2-one (18), together with the expected adduct 17 a. Compound 18 is formed by a nucleophilic addition to a CO ligand in a preformed carbene complex. This is a new example of the rare attack of a nucleophile on a CO ligand in a Fischer carbene complex. Adducts 10 form seven-membered-ring carbene complexes 19 and 20 by intramolecular aminolysis. In contrast, reaction of alkynyl carbene complexes with 1,8-diaminonaphthalene under very mild conditions leads to 2-substituted perimidines 33 together with the corresponding ethoxymethylmetal carbene complex 32 through an unprecedented fragmentation process in a formal retro-Aumann reaction. PMID- 14562314 TI - Hydrogenation versus transfer hydrogenation of ketones: two established ruthenium systems catalyze both. AB - The established standard ketone hydrogenation (abbreviated HY herein) precatalyst [Ru(Cl)(2)((S)-tolbinap)[(S,S)-dpen]] ((S),(S,S)-1) has turned out also to be a precatalyst for ketone transfer hydrogenation (abbreviated TRHY herein) as tested on the substrate acetophenone (3) in iPrOH under standard conditions (45 degrees C, 45 bar H(2) or Ar at atmospheric pressure). HY works at a substrate catalyst ratio (s:c) of up to 10(6) and TRHY at s:c<10(4). Both produce (R)-1-phenylethan 1-ol ((R)-4), but the ee in HY are much higher (78-83 %) than in TRHY (4-62 %). In both modes, iPrOK is needed to generate the active catalysts, and the more there is (1-4500 equiv), the faster the catalytic reactions. The ee is about constant in HY and diminishes in TRHY as more iPrOK is added. The ketone TRHY precatalyst [Ru(Cl)(2)((S,S)-cyP(2)(NH)(2))] ((S,S)-2), established at s:c=200, has also turned out to be a ketone HY precatalyst at up to s:c=10(6), again as tested on 3 in iPrOH under standard conditions. The enantioselectivity is opposite in the two modes and only high in TRHY: with (S,S)-2, one obtains (R)-4 in up to 98 % ee in TRHY as reported and (S)-4 in 20-25 % ee in HY. iPrOK is again required to generate the active catalysts in both modes, and again, the more there is, the faster the catalytic reactions. The ee in TRHY are only high when 0.5-1 equivalents iPrOK are used and diminish when more is added, while the (low) ee is again about constant in HY as more iPrOK is added (0-4500 equiv). The new [Ru(H)(Cl)((S,S)-cyP(2)(NH)(2))] isomers (S,S)-9 A and (S,S)-9 B (mixture, exact structures unknown) are also precatalysts for the TRHY and HY of 3 under the same conditions, and (R)-4 is again produced in TRHY and (S)-4 in HY, but the lower ee shows that in TRHY (S,S)-9 A/(S,S)-9 B do not lead to the same catalysts as (S,S)-2. In contrast, the ee are in accord with (S,S)-9 A/(S,S)-9 B leading to the same catalysts as (S,S)-2 in HY. The kinetic rate law for the HY of 3 in iPrOH and in benzene using (S,S)-9 A/(S,S)-9 B/iPrOK or (S,S)-9 A/(S,S)-9 B/tBuOK is consistent with a fast, reversible addition of 3 to a five-coordinate amidohydride (S,S)-11 to give an (S,S)-11-substrate complex, in competition with the rate-determining addition of H(2) to (S,S)-11 to give a dihydride [Ru(H)(2)((S,S)-cyP(2)(NH)(2))] (S,S)-10, which in turn reacts rapidly with 3 to generate (S)-4 and (S,S)-11. The established achiral ketone TRHY precatalyst [Ru(Cl)(2)(ethP(2)(NH)(2))] (12) has turned out to be also a powerful precatalyst for the HY of 3 in iPrOH at s:c=10(6) and of some other substrates. Response to the presence of iPrOK is as before, except that 12 already functions well without it at up to s:c=10(6). PMID- 14562315 TI - Modulating charge-transfer interactions in topologically different porphyrin-C60 dyads. AB - Control over the interchromophore separation, their angular relationship, and the spatial overlap of their electronic clouds in several ZnP-C(60) dyads (ZnP=zinc porphyrin) is used to modulate the rates of intramolecular electron transfer. For the first time, a detailed analysis of the charge transfer absorption and emission spectra, time-dependent spectroscopic measurements, and molecular dynamics simulations prove quantitatively that the same two moieties can produce widely different electron-transfer regimes. This investigation also shows that the combination of ZnP and C(60) consistently produces charge recombination in the inverted Marcus region, with reorganization energies that are remarkably low, regardless of the solvent polarity. The time constants of electron transfer range from the mus to the ps regime, the electronic couplings from a few tens to several hundreds of cm(-1), and the reorganization energies remain below 0.54 eV and can be as low as 0.16 eV. PMID- 14562316 TI - Systematic construction of a monotetrahydrofuran-ring library in annonaceous acetogenins by asymmetric alkynylation and stereodivergent tetrahydrofuran-ring formation. AB - All eight diastereoisomers of the monotetrahydrofuran-ring cores of annonaceous acetogenins have been synthesized through utilization of asymmetric alkynylation and stereodivergent one-pot tetrahydrofuran-ring formation. In all cases, the asymmetric alkynylation proceeded with very high diastereoselectivity to give eight kinds of optically pure tetrahydrofuran core from a common alpha oxyaldehyde. We also describe a comparison of the (1)H NMR, (13)C NMR, and CD spectral data of the eight isomers and give full details of the tetrahydrofuran ring construction including a model study of asymmetric alkynylation. PMID- 14562317 TI - Fabrication of self-supported patterns of aligned beta-FeOOH nanowires by a low temperature solution reaction. AB - Self-supported patterns of oriented alignment of beta-FeOOH nanowires are fabricated through a simple solution reaction from the complex [Fe(phen)(3)](2+) at 60 degrees C. The alignment of nanowires with a diameter of 40 nm and length of 6 mum is relatively uniform. HRTEM studies show that the growing direction of beta-FeOOH nanowires is perpendicular to the orientation plane of self-formed beta-FeOOH flake-like substrates. In the reaction and crystal growth process, the precursor [Fe(phen)(3)](2+) is undoubtedly vital to the formation of nanowire alignment. In detail, the formation of aligned nanowires is thought to be realized by controlling two competing reactions. Electrochemical and UV-visible measurements suggest that the product might have potential applications in lithium batteries and semiconductor electronics. This synthetic process is simple, mild, clean, reproducible, and free of any template; it provides a novel pathway for the low-temperature growth of nanowires and their simultaneous oriented alignment. PMID- 14562318 TI - The effect of ring nitrogen atoms on the homolytic reactivity of phenolic compounds: understanding the radical-scavenging ability of 5-pyrimidinols. AB - Six substituted 5-pyrimidinols were synthesized, and the thermochemistry and kinetics of their reactions with free radicals were studied and compared to those of equivalently substituted phenols. To assess their potential as hydrogen-atom donors to free radicals, we measured their O-H bond dissociation enthalpies (BDEs) using the radical equilibration electron paramagnetic resonance technique. This revealed that the O-H BDEs in 5-pyrimidinols are, on average, about 2.5 kcal mol(-1) higher than those in equivalently substituted phenols. The results are in good agreement with theoretical predictions, and confirm that substituent effects on the O-H BDE of 5-pyrimidinol are essentially the same as those on the Obond;H BDE in phenol. The kinetics of the reactions of these compounds with peroxyl radicals has been studied by their inhibition of the AIBN-initiated autoxidation of styrene, and with alkyl and alkoxyl radicals by competition kinetics. Despite their larger O-H BDEs, 5-pyrimidinols appear to transfer their phenolic hydrogen atom to peroxyl radicals as quickly as equivalently substituted phenols, while their reactivity toward alkyl radicals far exceeds that of the corresponding phenols. We suggest that this rate enhancement, which is large in the case of alkyl radical reactions, small in the case of peroxyl radical reactions, and nonexistent in the case of alkoxyl radical reactions, is due to polar effects in the transition states of these atom-transfer reactions. This hypothesis is supported by additional experimental and theoretical results. Despite this higher reactivity of 5-pyrimidinols towards radicals compared to phenols, electrochemical measurements indicate that they are more stable to one-electron oxidation than equivalently substituted phenols. For example, the 5-pyrimidinol analogues of 2,4,6-trimethylphenol and butylated hydroxytoluene (BHT) were found to have oxidation potentials approximately 400 mV higher than their phenolic counterparts, but reacted roughly one order of magnitude faster with alkyl radicals and at about the same rate with peroxyl radicals. The 5-pyrimidinol structure should, therefore, serve as a useful template for the rational design of novel air-stable radical scavengers and chain-breaking antioxidants that are more effective than phenols. PMID- 14562319 TI - Synthesis of a hexagonal nanosized macrocyclic fluorophore with integrated endotopic terpyridine metal-chelation sites. AB - A rigid nanosized hexagonal phenylethynyl cyclophane 5 has been prepared, which incorporates two 2,2':6',2"-terpyridines as integral structural units, for the purpose of binding metal ions. Macrocycle 5 was obtained by a 14-step synthesis in an overall yield of 11 %, and was characterised by spectroscopic techniques. The efficiency and ease of all transformations, and the relatively enhanced yield of the final macrocyclisation suggests that the entire synthetic pathway should be amenable to scale-up. Cyclophane 5 possesses four bulky triisopropylsilyl(TIPS)-protected ethyne substituents which serve a dual role. Firstly, they solubilise the structure thereby facilitating purification and subsequent handling. Secondly, they enable post-synthetic modification in which additional functionality may be attached to the periphery of the ring. Significantly, 5 was found to be a fluorescent chromophore, and may therefore potentially function as a new sensory platform for the detection of metal ions and H-bond donating biological substrates. The structurally well-defined nanosized morphology of 5, coupled with its interesting spectroscopic properties, supports the expectation that 5 and related architectures will attain a wealth of future applications within the developing fields of nanochemistry and nanoscience. PMID- 14562320 TI - The evolution of [[Ph2P(CH2)nPPh2]Pt(mu-S)2Pt[Ph2P(CH2)nPPh2]] (n=2, 3) metalloligands in protic acids: a cascade of sequential reactions. AB - Given the nucleophilicity of the [Pt(2)S(2)] ring, the evolution of [Pt(2)(mu S)(2)(P intersection P)(2)] (P intersection P=1,2-bis(diphenylphosphino)ethane (dppe), 1,3-bis(diphenylphosphino)propane (dppp)) metalloligands in the presence of the simplest electrophilic species, the proton, has been studied. Combined use of experimental and theoretical data has allowed the whole set of reactions ensuing the protonation of the [Pt(2)S(2)] core to be established. The titration of [Pt(2)(mu-S)(2)(P intersection P)(2)] with HCl or HClO(4) was monitored mainly by (31)P[(1)H] NMR and mass techniques. Characterization of all the species involved was completed with the determination of the crystal structure of [Pt(SH)(2)(P intersection P)], for dppe and dppp, and [Pt(3)(mu(3) S)(2)(dppp)(3)](PF(6))(2). The first protonation step of the [Pt(2)S(2)] core leads to the stable [Pt(2)(mu-S)(mu-SH)(P intersection P)(2)](+) complex, but the second step implies disintegration of the ring, thus giving rise to various mononuclear species. The subsequent evolution of some of these species allows regeneration of [Pt(2)(mu-S)(mu-SH)(P intersection P)(2)](+), evidencing the cyclic nature of this process. Whereas the reaction pathway is essentially common for both phosphine ligands, dppe and dppp, the different coordinating ability of Cl(-) or ClO(4) (-) determines the nature of the final products, [PtCl(2)(P intersection P)], [Pt(3)(mu(3)-S)(2)(P intersection P)(3)]Cl(2) or [Pt(3)(mu(3) S)(2)(P intersection P)(3)](ClO(4))(2). DFT calculations have corroborated the thermodynamic feasibility of the reactions proposed on the basis of experimental data. PMID- 14562321 TI - Synthesis and supramolecular properties of trimethylene-bridged clips. AB - The novel trimethylene-bridged clips 3 and 4 have been synthesized by using repetitive stereoselective Diels-Alder reactions of the benzo- and naphthobismethylenenorbornenes 8 and 19 as dienes and norbornadiene 9 as bisdienophile, and subsequent dehydrogenation of the primary cyclobisadducts 10 and 20 by using 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ). Clips 3 and 4 serve as receptors for a variety of electron-deficient neutral and cationic aromatic substrates, comparable to the molecular tweezers 1 and 2. The thermodynamic parameters of the complex formation, K(a) and DeltaG, were determined by (1)H NMR titration experiments and, in the case of the highly stable complex TCNB 32@4, by the use of isothermal titration microcalorimetry. The finding that clip 4 forms more stable complexes than 3 can be explained by the larger van der Waals contact surfaces of the naphthalene sidewalls in 4 compared to the corresponding benzene systems in 3. In the complexes with 4 as receptor, the plane of each aromatic substrate molecule is calculated to be oriented almost parallel to the naphthalene sidewalls. However, in the complexes of tweezers 2, the substrate is usually oriented parallel to the central naphthalene spacer unit. Due to the more open topology of 4, most complexes were calculated to consist of two or more equilibrating noncovalent conformers. PMID- 14562322 TI - Chirality transfer from guest chiral metal complexes to inorganic framework: the role of hydrogen bonding. AB - The structure elucidation of a new zinc phosphate [Co(II)(en)(3)][Zn(4)(H(2)PO(4))(3)(HPO(4))(2)(PO(4))(2 H(2)O)(2)] (1) reveals that the racemic cobalt complex templates the zinc phosphate framework in such a way that the local C(2) point symmetry of the structural motif of the inorganic framework conforms with that of the cobalt complex pairing with it, in essence transferring its chirality to the inorganic host. An analysis of hydrogen bonding between the guest molecules and the inorganic host framework reveals that hydrogen bonding is responsible for the stereospecific structural arrangement. Upon examining previously reported chiral metal-complex-templated structures of metal phosphates, it is revealed that such hydrogen bonding is the common origin for inducing chirality transfer in metal-phosphate frameworks templated with chiral metal complexes. Crystal data of 1: orthorhombic, Pbcn (no. 60), a=10.4787(8) A, b=20.0091(14) A, c=14.9594(10) A, and Z=2. PMID- 14562323 TI - Aromaticity and electron affinity of Carbo(k)-[3]radialenes, k=0, 1, 2. AB - Aromaticity enhancement is a possible driving force for the low reduction potentials of buta-1,3-diynediyl-expanded [N]radialenes: this hypothesis is theoretically analyzed for the expanded [3]radialene prototype. This study is undertaken within a more general prospect, namely the evaluation of the variation of aromaticity with endocyclic and peripheral carbomeric expansion of [3]radialene and its mono- and dianions. The structures, denoted as [C-H](6) (h)[C-C](3) (k)carbo-[3]radialene(q) (h=0, 1; k=0, 1, 2; q=0, -1, -2), were optimized in relevant singlet, doublet, or triplet spin states at the B3PW91/6 31G** level. They were found to be all planar. The structural aromaticity was measured through the average bond length d(av) over the [C-C](3) (k)carbo [3]radialene core, and by the corresponding bond-length equalization parameter sigma(d), related to Krygowski's GEO. The magnetic aromaticity was measured by Schleyer's NICS values at the center of the rings. Regarding the relative variation of NICS and sigma(d), two classes of species can be distinguished according to their endocyclic expansion level. The species with a nonexpanded (k=0) or doubly expanded (k=2) ring constitute the first class: they exhibit D(3h) symmetry and a strong correlation of NICS with sigma(d). The species with a singly expanded ring (k=1) fall far from the correlation line, and constitute the second class. This class distinction is related to the degeneracy scheme of the frontier orbitals of the neutral representative. A finer appraisal of the electron (de)localization is brought by the ELF (Electron Localization Function) analysis of the electron density. It allows for a weighting of relevant resonance forms. Unsubstituted species are well described by the superimposition of two or three resonance forms. For (doublet spin state) monoanionic species, their respective weights are validated by comparison with AIM spin density. The weighted mean, n, of the formal numbers of paired pi(z) electrons in the resonance forms was calculated and compared with the closest even integer of either forms 4m+2 or 4m. A density-based continuous generalization of the orbital based discrete Huckel rule is then heuristically proposed through an analytical correlation of NICS versus sigma(d), n, and S, the spin of the species. The frontier-orbital-degeneracy pattern of neutral species is discussed with respect to structural and magnetic aromaticity criteria. A decreasing HOMO-LUMO gap versus endocyclic expansion is obtained, but [C-C](3) (1)carbo-[3]radialene possesses the highest HOMO and LUMO energies. Vertical and adiabatic electron affinities of neutral and monoanionic species were also computed and compared with related experimental data. PMID- 14562324 TI - Tailoring the catalytic performance of sol-gel-encapsulated tetra-n propylammonium perruthenate (TPAP) in aerobic oxidation of alcohols. AB - Sol-gel nanohybrid silica particles organically modified and doped with the ruthenium species tetra-n-propylammonium perruthenate (TPAP) are highly efficient catalysts for the selective oxidation of alcohols to carbonyl groups with O(2) at low pressure in toluene. The materials are easily prepared by a one-step sol-gel process, and their catalytic performance can be optimised by tailoring the conditions of their synthesis by hydrolytic co-polycondensation of tetramethoxysilane (TMOS) and alkyltrimethoxysilanes R-Si(OMe)(3) in the presence of TPAP. Eventually, heterogeneous catalysts considerably more active than the unsupported perruthenate were obtained, while also being leach-proof and recyclable. The correlation between the materials' activity, surface polarity and textural properties suggests valuable information on the chemical behaviour of sol-gel catalytic materials in oxidation catalysis; this is of interest in view of the importance of efficient solid catalysts for the selective oxidation of alcohols with O(2). PMID- 14562325 TI - Trivalent boron as an acceptor in donor-pi-acceptor-type compounds for single- and two-photon excited fluorescence. AB - The synthesis, structure, and fluorescence properties of a series of new donor-pi acceptor (D-pi-A) type compounds, with a trivalent boron, protected by two mesityl groups, as acceptor, and with various typical donors and different pi conjugated bridges, are reported. All these stable organoboron compounds show intense single-photon excited fluorescence (SPEF) and two-photon excited fluorescence (TPEF) in a wide spectral range from blue to green, with the spectral peak position of the SPEF being basically the same as that of the TPEF. The remarkably strong Cbond;B(mesityl)(2) bonding, and the well-conjugated pi system, shown in X-ray crystal structures of two compounds, indicate some charge transfer features of the ground state. Meanwhile, spectral data indicate that the charge transfer from donor to acceptor is greatly enhanced in the excited states. Based on typical structural data and comprehensive spectral data, the following structure-property relationships can be drawn: 1) the moderate arylamino donor can more effectively enhance the SPEF and TPEF intensities than can the strong alkylamino donor; 2) stilbene is a better pi-bridge than styrylthiophene for its capability of enhancing and blue-shifting the SPEF and TPEF of the corresponding D-pi-A compounds; and 3) when compared to its boron-free precursors and other analogues, -B(mesityl)(2) invariably and consistently acts as an effective SPEF and TPEF fluorophore in all this series of organoboron compounds, which may result from its strong pi-electron-withdrawing and charge transfer-inducing nature in the ground-state and, more dominantly, in the excited-state. Combining all the above positive structure factors, trans-4'-N,N-diphenylamino-4 dimesitylborylstilbene (compound 3) stands out as the optimized green SPEF and TPEF emitter. This compound exhibits an SPEF quantum yield Phi of 0.91 at 522 nm in THF, a TPEF cross-section sigma' that is an order of magnitude larger than that of its boron-free precursor upon excitation by 800 nm femto-second laser pulses, and a two-photon absorption section sigma of 3.0 x 10(-48) cm(4) s. In the blue light region, trans-4'-N-carbazolyl-4-dimesitylboryl-stilbene (compound 4) shows significant SPEF and TPEF properties, with Phi=0.79 at 464 nm in THF and a large sigma' value, which is five times that of fluorescein upon excitation by 740 nm femto-second laser pulses. PMID- 14562326 TI - Bisporphyrins with bischlorin features obtained by direct anodic coupling of porphyrins. AB - The proximity of two or more porphyrin or chlorin-like structures has been shown to be crucial in numerous biological processes, such as electron transfers. The one-pot electrochemical synthesis of a dimeric tetraphenylporphyrin with one 1,2 (diphenylphosphonium)benzene as a spacer and of its porphyrin-monomer precursor are reported. These new compounds have been characterized by (1)H and (31)P NMR, ESR, and UV-visible spectroscopy and microanalysis. Electrochemical data are reported, and the redox behavior is analyzed for the monomers and the dimers. Important interactions between the two chromophores and a phosphonium-phosphonium interaction have been observed. UV-visible and (1)H NMR data along with electrochemical behavior suggest that the positive charge carried by the two phosphonium units is in part delocalized onto the pi system of the porphyrins, this gives an unexpected bis-porphyrin with bischlorin spectroscopic features. PMID- 14562328 TI - Discrimination of peptides by using a molecularly imprinted piezoelectric biosensor. AB - Based on the direct formation of a molecularly imprinted polymer on gold electrodes, we have developed a peptide sensor for the detection of low-molecular weight peptides. A new cross-linking monomer, (N-Acr-L-Cys-NHBn)(2), was employed to attach the surface of the chip and to copolymerize with other monomers. Interestingly, N-benzylacrylamide participates in the polymerization and recognition is carried out in an aqueous environment. By using quartz crystal microbalance detection, short peptides can be monitored by their interaction with plastic antibodies specific for the target peptides. The selectivity of molecularly imprinted polymers and the sensitivity of such artificial biosensors have been combined to differentiate between traces of oxytocin and vasopressin to the ng mL(-1) scale. PMID- 14562329 TI - Remarkable reactions of cyclooctatetraene diiron-bridging carbyne complexes with amino and amido compounds: nucleophilic addition to and breaking of the cyclooctatetraene ring. AB - The reactions of the cationic, diiron-bridging carbyne complexes [Fe(2)(mu CAr)(CO)(4)(eta(8)-C(8)H(8))]BF(4) (1, Ar=C(6)H(5); 2, Ar=p-CH(3)C(6)H(4); 3, Ar=p-CF(3)C(6)H(4)) with LiN(C(6)H(5))(2) in THF at low temperature gave novel N nucleophilic-addition products, namely, the neutral, diiron-bridging carbyne complexes [Fe(2)(mu-CAr)(CO)(4)(eta(7)-C(8)H(8)N(C(6)H(5))(2))] (4, Ar=C(6)H(5); 5, Ar=p-CH(3)C(6)H(4); 6, Ar=p-CF(3)C(6)H(4))). Cationic bridging carbyne complexes 1-3 react with (C(2)H(5))(2)NH, (iC(3)H(7))(2)NH, and (C(6)H(11))(2)NH under the same conditions with ring cleavage of the COT ligand to produce the novel diiron-bridging carbene inner salts [Fe(2)[mu-C(Ar)C(8)H(8)NR(2)](CO)(4)] (7, Ar=C(6)H(5), R=C(2)H(5); 8, Ar=p-CH(3)C(6)H(4), R=C(2)H(5); 9, Ar=p CF(3)C(6)H(4), R=C(2)H(5); 10, Ar=C(6)H(5), R=iC(3)H(7); 11, Ar=p-CH(3)C(6)H(4), R=iC(3)H(7); 12, Ar=p-CF(3)C(6)H(4), R=iC(3)H(7); 13, Ar=C(6)H(5), R=C(6)H(11); 14, Ar=p-CH(3)C(6)H(4), R=C(6)H(11), 15, Ar=p-CF(3)C(6)H(4), R=C(6)H(11)). Piperidine reacts similarly with cationic carbyne complex 3 to afford the corresponding bridging carbene inner salt [Fe(2)[mu C(Ar)C(8)H(8)N(CH(2))(5)](CO)(4)] (16). Compound 9 was transformed into a new diiron-bridging carbene inner salt 17, the trans isomer of 9, by heating in benzene. Unexpectedly, the reaction of C(6)H(5)NH(2) with 2 gave a novel COT iron carbene complex [Fe(2)[=C(C(6)H(4)CH(3)-p)NHC(6)H(5)](mu-CO)(CO)(3)(eta(8) C(8)H(8))] (18). However, the analogous reactions of 2-naphthylamine with 2 and of p-CF(3)C(6)H(4)NH(2) with 3 produce novel chelated iron-carbene complexes [Fe(2)[=C(C(6)H(4)CH(3)-p)NC(10)H(7)](CO)(4)(eta(2):eta(3):eta(2)-C(8)H(9))] (19) and [Fe(2)[=C(C(6)H(4)CF(3)-p)NC(6)H(4)CF(3)-p](CO)(4)(eta(2):eta(3):eta(2) C(8)H(9))] (20), respectively. Compound 18 can also be transformed into the analogous chelated iron-carbene complex [Fe(2)[=C(C(6)H(4)CH(3) p)NC(6)H(5)](CO)(4)(eta(2):eta(3):eta(2)-C(8)H(9))] (21). The structures of complexes 6, 9, 15, 17, 18, and 21 have been established by X-ray diffraction studies. PMID- 14562327 TI - Transition-metal complexes as enzyme-like reagents for protein cleavage: complex cis-[Pt(en)(H2O)2]2+ as a new methionine-specific protease. AB - Complex cis-[Pt(en)(H(2)O)(2)](2+) promotes selective hydrolytic cleavage of two proteins, horse cytochrome c and bovine beta-casein. The cleavage is completed in 24 h under relatively mild conditions, at about pH 2.5, and a temperature as low as 40 degrees C. The results of HPLC and TSDS PAGE separations, MALDI mass spectrometry, and Edman sequencing showed that cleavage occurred exclusively at the peptide bond involving the C-terminus of each methionine residue, both such residues in cytochrome c and all six such residues in beta-casein. While having the same selectivity as cyanogen bromide (CNBr), the most common chemical protease, cis-[Pt(en)(H(2)O)(2)](2+) has several advantages. It is nonvolatile, easy to handle, and recyclable. Its cleavage is residue-selective, the rest of the polypeptide backbone remains intact, and the other side chains remain unmodified. It is applied in approximately equimolar amounts with respect to methionine residues, creates free amino and carboxylic groups, and cleaves even the Met-Pro bond, which is resistant to CNBr and most proteolytic enzymes. Finally the complex also works in the presence of the denaturing reagent sodium dodecyl sulfate. Experiments with the synthetic peptides, AcAla-Lys-Tyr-Gly-Gly Met-Ala-Ala-Arg-Ala (termed Met-peptide) and AcVal-Lys-Gly-Gly-His-Ala-Lys-Tyr Gly-Gly-Met-Ala-Ala-Arg-Ala (termed HisMet-peptide) as substrates, revealed structural and mechanistic features of the proteolytic reactions. We explain why two similar complexes with similar metal ions, cis-[Pt(en)(H(2)O)(2)](2+) and cis [Pd(en)(H(2)O)(2)](2+), differ in selectivity as proteolytic reagents. The selectivity of cleavage is governed by the selectivity of the cis [Pt(en)(H(2)O)(2)](2+) binding to the methionine side chain. The proteolytic activity is governed by the modes of coordination, which control the approach of the anchored Pt(II) ion to the scissile peptide bond. The cleavage occurs with a small, but significant, catalytic turnover of more than 18 after 7 days. The ability of cis-[Pt(en)(H(2)O)(2)](2+) to cleave proteins at relatively few sites, with explicable selectivity and catalytic turnover, bodes well for its use in biochemical practice. PMID- 14562330 TI - Effect of peripheral substitution on the electronic absorption and fluorescence spectra of metal-free and zinc phthalocyanines. AB - The effect of substituents on the position and intensity of the electronic absorption and fluorescence spectra of phthalocyanines (Pcs) was examined for 35 Pc compounds. When electron-releasing groups are bound to four alpha-benzo positions of the Pc skeleton, the B and Q bands shift to longer wavelength. Relative to this shift, the effect of introducing the same electron-releasing groups at the other four alpha positions amounts to about 1.6-2.0. Although the effect is not always clearly seen, introduction of electron-releasing groups in the beta-benzo positions of the Pc skeleton generally shifts the Q band to shorter wavelength. The effect of electron-withdrawing groups is exactly the opposite with respect to the alpha and beta positions. These effects can be reasonably explained by considering the magnitude of the atomic orbital coefficients of the carbon atoms derived from molecular orbital (MO) calculations. In addition, the following intriguing phenomena were observed in the experiments, although not all were explained theoretically: 1) the splitting of the Q band of metal-free Pcs decreases with increasing wavelength of the Q band, 2) the ring currents of Pcs with Q bands at longer wavelength are generally smaller, and 3) the absorption coefficients of the Q band of Pc compounds with 16 electron-releasing substituents are larger than those of the corresponding tetra- and octasubstituted Pcs by several tens of percent. 4) Our PPP calculations suggested that the absorption coefficient of the Q band of Pcs with more strongly electron releasing substituents is larger. 5) The second HOMO of the Pcs with the Q band at longer wavelength has b(1u) symmetry, as opposed to the a(2u) symmetry of normal Pcs. 6) Pcs showing S1 emission maxima at wavelengths longer than about 740 nm generally have quantum yields of less than 0.1. PMID- 14562331 TI - The open-chain trioxide CF3OC(O)OOOC(O)OCF3. AB - The open-chain trioxide CF(3)OC(O)OOOC(O)OCF(3) is synthesised by a photochemical reaction of CF(3)C(O)OC(O)CF(3), CO and O(2) under a low-pressure mercury lamp at -40 degrees C. The isolated trioxide is a colourless solid at -40 degrees C and is characterised by IR, Raman, UV and NMR spectroscopy. The compound is thermally stable up to -30 degrees C and decomposes with a half-life of 1 min at room temperature. Between -15 and +14 degrees C the activation energy for the dissociation is 86.5 kJ mol(-1) (20.7 kcal mol(-1)). Quantum chemical calculations have been performed to support the vibrational assignment and to discuss the existence of rotamers. PMID- 14562332 TI - Synthetic and structural studies of cobalt-pivalate complexes. AB - The synthesis and characterisation of a range of cobalt pivalate cage complexes are reported. The cages include: a dinuclear Co(II) complex; an oxo-centred Co(III) triangle; tetranuclear Co(II) heterocubanes and butterflies; tetranuclear heterovalent cobalt butterflies and hexanuclear edge-sharing bitetrahedra; heterovalent penta-, hexa- and hepta-nuclear cages based on [M(4)O(4)] heterocubane cores; and a tetradecanuclear cage based on heterocubanes sharing edges and vertices. Spectroscopic studies suggest that some of these cores are retained in solution, but that only in the Co(III) triangle is the structure including ligands retained. A scheme is proposed to account for the many structures observed, which may be applicable to other polymetallic cage complexes. PMID- 14562338 TI - Improved chemical strategies for the targeted therapy of cancer. PMID- 14562339 TI - The arduous way to the egg: follow the nose. PMID- 14562340 TI - Recent advances in the activation of boron and silicon reagents for stereocontrolled allylation reactions. AB - Despite the popularity of boron and silicon allylation reagents in stereocontrolled synthesis, they suffer from a number of inherent limitations that have slowed down their development as synthetic tools for nucleophilic additions to carbonyl compounds and imine derivatives. These limitations are the low reactivity and diastereoselectivity of allyl trialkylsilane reagents, and the lack of catalytic systems for the activation and substoichiometric control of enantioselectivity in the additions of allyl boron reagents. To develop more efficient and general methods for the control of absolute stereochemistry in the resulting homoallylic alcohols, new approaches aimed at solving the problem of activation of allylic boron and silicon reagents are needed. This Minireview describes a number of recent approaches that have been devised to address this problem. PMID- 14562341 TI - Sulfur and selenium: the role of oxidation state in protein structure and function. AB - Sulfur and selenium occur in proteins as constituents of the amino acids cysteine, methionine, selenocysteine, and selenomethionine. Recent research underscores that these amino acids are truly exceptional. Their redox activity under physiological conditions allows an amazing variety of posttranslational protein modifications, metal free redox pathways, and unusual chalcogen redox states that increasingly attract the attention of biological chemists. Unlike any other amino acid, the "redox chameleon" cysteine can participate in several distinct redox pathways, including exchange and radical reactions, as well as atom-, electron-, and hydride-transfer reactions. It occurs in various oxidation states in the human body, each of which exhibits distinctive chemical properties (e.g. redox activity, metal binding) and biological activity. The position of selenium in the periodic table between the metals and the nonmetals makes selenoproteins ideal catalysts for many biological redox transformations. It is therefore apparent that the chalcogen amino acids cysteine, methionine, selenocysteine, and selenomethionine exhibit a unique biological chemistry that is the source of exciting research opportunities. PMID- 14562342 TI - Design and total synthesis of a superior family of epothilone analogues, which eliminate xenograft tumors to a nonrelapsable state. PMID- 14562345 TI - One-pot synthesis of polyheterocycles by a palladium-catalyzed intramolecular N arylation/C[bond]H activation/aryl[bond]aryl bond-forming domino process. PMID- 14562344 TI - Asymmetric hetero-ene reactions of trimethylsilyl enol ethers catalyzed by tridentate Schiff base chromium(III) complexes. PMID- 14562343 TI - Trapping highly electrophilic metalladiphosphanylcarbenes. PMID- 14562346 TI - Substituted 1,4-bisallylphosphonates: preparation from 1-alkynylphosphonates and Ti(OiPr)4/2 iPrMgCl. PMID- 14562347 TI - Total synthesis of natural (+)-lasonolide A. PMID- 14562350 TI - CuH-catalyzed asymmetric conjugate reductions of acyclic enones. PMID- 14562348 TI - A thiazyl-based organic ferromagnet. PMID- 14562349 TI - Surface and site-specific ring-opening metathesis polymerization initiated by dip pen nanolithography. PMID- 14562351 TI - Catalytic enantioselective conjugate reduction of beta,beta-disubstituted nitroalkenes. PMID- 14562352 TI - Time scale and elementary steps of CO-induced disintegration of surface rhodium clusters. PMID- 14562353 TI - A new route to crystalline hydrogels, guided by a phase diagram. PMID- 14562354 TI - A fluorescent self-amplifying wavelength-responsive sensory polymer for fluoride ions. PMID- 14562355 TI - Directed evolution of an amine oxidase possessing both broad substrate specificity and high enantioselectivity. PMID- 14562356 TI - Structure and magnetic properties of a chiral two-dimensional ferrimagnet with TC of 38 K. PMID- 14562357 TI - Comprehensive step-by-step engineering of an (R)-hydroxynitrile lyase for large scale asymmetric synthesis. PMID- 14562358 TI - Cs2Pt: a platinide(-II) exhibiting complete charge separation. PMID- 14562359 TI - Stereospecific biocatalytic synthesis of pancratistatin analogues. PMID- 14562360 TI - [2+2] photocycloaddition of tetronates. PMID- 14562361 TI - Enhancing the catalytic activity of 4-(dialkylamino)pyridines by conformational fixation. PMID- 14562363 TI - Gastroesophageal reflux disease at the turn of millennium. AB - Gastroesophageal reflux disease (GERD) has been an area of active research in the Asia-Pacific region in the recent years. This article outlines some of the interesting research findings. It comprises three parts. The first part dealt with recent data on the changing epidemiology of GERD in Asia. The second part summarized published studies on the relationship between GERD and Helicobacter pylori, relevant to the Asia-Pacific region. The last part discussed some of the recent advances in the treatment of GERD. PMID- 14562364 TI - Helicobacter pylori infection and micronutrient deficiencies. AB - It is known that deficiencies of micronutrients due to infections increase morbidity and mortality. This phenomenon depicts itself conspicuously in developing countries. Deficiencies of iron, vitamins A, E, C, B12, etc are widely prevalent among populations living in the third world countries. Helicobacter pylori (H pylori) infection has a high prevalence throughout the world. Deficiencies of several micronutrients due to H pylori infection may be concomitantly present and vary from subtle sub-clinical states to severe clinical disorders. These essential trace elements/micronutrients are involved in host defense mechanisms, maintaining epithelial cell integrity, glycoprotein synthesis, transport mechanisms, myocardial contractility, brain development, cholesterol and glucose metabolism. In this paper H pylori infection in associated with various micronutrients deficiencies is briefly reviewed. PMID- 14562365 TI - Pathophysiological aspects of diverticular disease of colon and role of large bowel motility. AB - Colonic diverticular disease (diverticulosis) is one of the most common gastrointestinal disorders in Western countries. This disorder is strictly related to aging and fibre intake, and still bears a discrete amount of morbidity. Numerous etiological co-factors have to date been implicated in the pathogenesis of the disease, yet the supporting evidence is still far from absolute. The present review considers the pathophysiology of colonic diverticular disease, with a special emphasis on factors related to abnormal colonic motility. PMID- 14562366 TI - Separation and identification of differentially expressed nuclear matrix proteins between human esophageal immortalized and carcinomatous cell lines. AB - AIM: To separate and identify differentially expressed nuclear matrix proteins (NMPs) between the immortalized human esophageal epithelial cell line (SHEE) and the malignantly transformed esophageal carcinoma cell line (SHEEC), and to provide new ways for finding specific markers and the pathogenesis of esophageal carcinoma. METHODS: SHEE and SHEEC cell lines were used to extract NMPs. The quality of NMPs was monitored by Western blot analysis including DNA topoisomerase IIalpha, proliferation cell nuclear antigen (PCNA) and histone. NMPs of SHEE and SHEEC were analyzed by two-dimensional electrophoresis (2-DE), silver staining and PDQuest6.2 image analysis software. Three spots in which the differentially expressed NMPs were more obvious, were selected and analyzed with matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI- TOF-MS) and database search. RESULTS: Western blot analysis revealed that DNA topoisomerase IIalpha and PCNA were detected, and the majority of histones were deleted in NMPs of SHEE and SHEEC. After 2-DE image analysis by PDQuest6.2 software, the 2-DE maps were detected with an average of 106+/-7.1 spots in SHEE and 132+/-5.0 spots in SHEEC. Most of them were matched one another (r=0.72), only 16 protein spots were found differing in intensity. Three NMPs including cytoskeletal tropomyosin, FK506-binding protein 6, similar to retinoblastoma binding protein 8 were preliminarily identified by MALDI- TOF-MS. CONCLUSION: These differentially expressed NMPs may play an important role during malignant transformation from SHEE to SHEEC. Their separation and identification will contribute to searching for specific markers and probing into the pathogenesis of esophageal carcinoma. PMID- 14562367 TI - 15d-PGJ2 inhibits cell growth and induces apoptosis of MCG-803 human gastric cancer cell line. AB - AIM: To investigate the influence of peroxisome proliferator-activated receptor gamma (PPARgamma) ligand, 15-deoxy-12, 14-prostaglandin J2 (15dPGJ2) on the proliferation and apoptosis of MCG-803 human gastric cancer cell lines. METHODS: Cell proliferation was measured by 3H-TdR assay. Apoptosis was determined by ELISA and TUNEL staining. Protein and mRNA level of bcl-2 family and COXs were measured by Western blotting and Northern blotting respectively. PGE2 production was examined by RIA. RESULTS: 15dPGJ2 inhibited cell growth and induced apoptosis of MCG-803 cells. The COX-2 and bcl-2/bax ratios were decreased following 15dPGJ2 treatment. The PGE2 production in supernatants was also decreased. These changes were in a dose-dependent manner. CONCLUSION: 15dPGJ2 may be a useful therapeutic agent for the treatment of gastric cancer. PMID- 14562368 TI - Ets1 as a marker of malignant potential in gastric carcinoma. AB - AIM: Ets1 proto-oncogene is a transcription factor involved in the activation of several genes of tumor invasion and metastasis. We aimed to determine the relationship between the extent and intensity of Ets1 expression and patients' clinicopathological factors in gastric carcinoma. METHODS: Immunohistochemical analysis was performed for gastric tumor paraffin-embedded sections, followed by image analysis. RESULTS: Ets1 was not expressed in the normal gastric epithelium and its surrounding cells. The percentage of Ets1 expressing cells detected increased significantly in both epithelial tumor and stromal cells from high T classification, lymph node metastasis positive, clinical advanced-stage groups (P<0.001). The level of Ets1 staining in epithelial tumor cells also reflected the degree of cell differentiation. The percentage of epithelial and stromal cells expressing Ets1 was significantly correlated with the presence of lymph node metastasis (P=0.014 and P<0.001 respectively). Ets1 expression was not observed in tissue samples from patients with benign gastric ulcers. CONCLUSION: Ets1 protein expression in epithelial tumor cells reflects the degree of differentiation, and the percentage of Ets1 positive tumor and stromal cells correlates with lymph node metastasis. Thus Ets1 is a valuable marker of malignant potential in terms of invasiveness and metastasis of gastric carcinoma. It is also possible that inhibition of Ets1 is a potential avenue for therapy in gastric cancer. PMID- 14562369 TI - Expression of MTLC gene in gastric carcinoma. AB - AIM: To investigate the expression of c-myc target from laryngeal cancer cells (MTLC) gene in gastric carcinoma (GC) tissues and the effect of MTLC over expression on gastric carcinoma cell line BGC823. METHODS: RT-PCR was performed to determine the expression of MTLC mRNA in GC and matched control tissues. BGC823 cells were transfected with an expression vector pcDNA3.1-MTLC by liposome and screened by G418. Growth of cells expressing MTLC was observed daily by manual counting. Apoptotic cells were determined by TdT-mediated dUTP nick-end labeling (TUNEL) assay. RESULTS: The expression of MTLC mRNAs was down-regulated in 9(60%) of 15 cases of GC tissues. The growth rates of the BGC823 cells expressing MTLC were indistinguishable from that of control cells. A marked acceleration of apoptosis was observed in MTLC-expressing cells. CONCLUSION: MTLC was down-regulated in the majority of GC tissues and could promote apoptosis of GC cell lines, which suggests that MTLC may play an important role in the carcinogenesis of gastric carcinoma. PMID- 14562370 TI - Identification of antigens by monoclonal antibody PD4 and its expression in Escherichia coli. AB - AIM: To clone and express the antigen of monoclonal antibody (MAb) PD4 for further investigation of its function. METHODS: MGC803 cDNA expression library was constructed and screened with PD4 as probes to clone the antigen. After failed in the library screening, immunoprecipitation and SDS-polyacrylamide gel electrophoresis were applied to purify the antigen for sequence analysis. The antigen coming from Mycoplasma hyorhinis (M. hyorhinis) was further confirmed with Western blot analysis by infecting M. hyorhinis -free HeLa cells and eliminating the M. hyorhinis from MGC803 cells. The full p37 gene was cloned by PCR and expressed successfully in Escherichia coli after site-directed mutations. Immunofluorescence assay was used to demonstrate if p37 protein could directly bind to gastric tumor cell AGS. RESULTS: The cDNA library constructed with MGC803 cells was screened by MAb PD4 as probes. Unfortunately, the positive clones identified with MAb PD4 were also reacted with unrelated antibodies. Then, immunoprecipitation was performed and the purified antigen was identified to be a membrane protein of Mycoplasma hyorhinis (M. hyorhinis) by sequencing of N terminal amino acid residues. The membrane protein was intensively verified with Western blot by eliminating M. hyorhinis from MGC803 cells and by infecting M. hyorhinis-free HeLa cells. The full p37 gene was cloned and expressed successfully in Escherichia coli after site-directed mutations. Immunofluorescence demonstrated that p37 protein could directly bind to gastric tumor cell AGS. CONCLUSION: The antigen recognized by MAb PD4 is from M. hyorhinis, which suggests the actions involved in MAb PD4 is possibly mediated by p37 protein or M. hyorhinis. As p37 protein can bind directly to tumor cells, the pathogenic role of p37 involved in tumorigenesis justifies further investigation. PMID- 14562371 TI - Different approaches to caudate lobectomy with "curettage and aspiration" technique using a special instrument PMOD: a report of 76 cases. AB - AIM: To study different approaches to caudate lobectomy with "curettage and aspiration" technique using Peng's multifunctional operative dissector (PMOD). The surgical procedure of isolated complete caudate lobectomy was specially discussed. METHODS: In 76 cases of various types of caudate lobectomy, three approaches were used including left side approach, right side approach, and anterior approach. Among the 76 cases, isolated complete caudate lobectomy was carried out in 6 cases with transhepatic anterior approach. The surgical procedure consisted of mobilization of the total liver, ligation and separation of the short hepatic veins, splitting the liver parenchyma through the Cantlie's plane, ligation and division of the caudate portal triads from the hilum, dissection of the root of major hepatic veins, detachment of the caudate lobe from liver parenchyma. RESULTS: The mean operative time was 285+/-51 min, the mean blood loss was 1,600 ml. No severe complications were observed. Among the 6 cases receiving isolated complete caudate lobectomy with transhepatic anterior approach, one case died 17 months after operation due to disease recurrence and liver failure, the other 5 cases have been alive without recurrence, with one longest survival of 49 months. CONCLUSION: The choice of approach is essential to the success of caudate lobectomy. As PMOD and "curettage and aspiration" technique can delineate intrahepatic or extrahepatic vessels clearly, caudate lobe resection has become safer, easier and faster. PMID- 14562372 TI - Inhibitory effects of antisense RNA of HAb18G/CD147 on invasion of hepatocellular carcinoma cells in vitro. AB - AIM: To study the inhibitory effects of antisense RNA of HAb18G/CD147 on invasion of hepatocellular carcinoma(HCC) cells in vitro. METHODS: Antisense RNA of HAb18G/CD147 vector PCI-asHAb18G was constructed by reversely inserting HAb18G/CD147 cDNA to eukaryotic expression vector PCI-neo. The HCC cell line HHCC was transfected by PCI-asHAb18G via cation liposome. Expression of HAb18G/CD147 of transfected cells selected by G418 (geneticin) was observed by immuno histochemical SP staining and FACS (fluorescence activated cell sorting). Gelatin zymography was used to determine the effect of PCI-asHAb18G on reducing secretions of MMP-2 and MMP-9 of the transfected cells. Boyden chamber was employed to test the invasion of HCC cells in vitro. RESULTS: The construction of antisense RNA vector PCI-asHAb18G was verified correct by partial nucleotide sequencing and restricted endonuclease digestion. The expression of HAb18G/CD147 in transfected HHCC was inhibited by PCI-asHAb18G. Secretions of MMP-2 and MMP-9 of transfected HHCC were reduced and the invasion of transfected HHCC was inhibited compared to HHCC, respectively. CONCLUSION: Invasion of HCC cells can be inhibited by antisense RNA of HAb18G/CD147. HAb18G/CD147 may be used as a potential target of drugs for anti-invasion and metastasis of HCC. PMID- 14562373 TI - Role of serum total sialic acid in differentiating cholangiocarcinoma from hepatocellular carcinoma. AB - AIM: This study was designed to evaluate the clinical application of serum total sialic acid (TSA) in the diagnosis of cholangiocarcinoma (CCA). METHODS: Serum TSA was determined by periodate-resorcinol microassay in 69 patients with CCA, 59 patients with hepatocellular carcinoma (HCC), 37 patients with cirrhosis, 61 patients with chronic hepatitis and 50 healthy blood donors. RESULTS: The mean serum TSA concentration in CCA (2.41+/-0.70 mmol/L) was significantly higher than those of HCC, cirrhosis, chronic hepatitis and healthy blood donors (1.41+/-0.37 mmol/L, 1.13+/-0.31 mmol/L, 1.16+/-0.26 mmol/L, and 1.10+/-0.14 mmol/L, respectively; P<0.001). Based on ROC curve analysis, a cut-off point of 1.75 mmol/L discriminated between CCA and HCC with a sensitivity, specificity and accuracy of 82.6 %, 83.1 %, and 82.8 %, respectively. CONCLUSION: Based on our results, serum TSA would be a useful marker for the differential diagnosis of CCA from HCC. PMID- 14562374 TI - Prevention of hepatocellular carcinoma in mice by IL-2 and B7-1 genes co transfected liver cancer cell vaccines. AB - AIM: To study the immunoprotective effect of liver cancer vaccine with co transfected IL-2 and B7-1 genes on hepatocarcinogenesis in mice. METHODS: The murine liver cancer cell line Hepal-6 was transfected with IL-2 and/or B7-1 gene via recombinant adenoviral vectors and the liver cancer vaccines were prepared. C57BL/6 mice were immunized with these vaccines and challenged with the parental Hepal-6 cells afterwards. The immunoprotection was investigated and the reactive T cell line was assayed. RESULTS: The immunoprotection of the tumor vaccine was demonstrated. The effect of IL-2 and B7-1 genes co-transfected Hepal-6 liver cancer vaccine (Hep6-IL2/B7 vaccine) on the onset of tumor formation was the strongest. When attacked with wild Hepal-6 cells, the median survival period of the mice immunized with Hep6-IL2/B7 vaccine was the longest (68 days, chi2=7.70 11.69, P<0.05) and the implanted tumor was the smallest (z =3.20-44.10, P<0.05). The effect of single IL-2 or B7-1 gene-transfected vaccine was next to the IL2/B7 gene co-transfected group, and the mean survival periods were 59 and 54 days, respectively. The mean survival periods of wild or enhanced green fluorescence protein gene modified vaccine immunized group were 51 and 48 days, respectively. The mice in control group all died within 38 days and the implanted tumor was the largest (z=3.20-40.21, P<0.05). The cellular immunofunction test and cytotoxicity study showed that the natural killer (NK) cell, lymphokine activated killer (LAK) cell and cytotoxic T lymphocyte (CTL) activities were significantly increased in mice immunized with the Hep6-IL2/B7 vaccine, (29.5+/-2.5 %, 65.0+/-2.9 %, 83.1+/ 1.5 % respectively, compared with other groups, P<0.05). CONCLUSION: The Hep6 IL2/B7 liver cancer vaccines can induce the mice to produce activated and specific CTL against the parental tumor cells, and demonstrate stronger effect on the hepatocarcinogenesis than single gene modified or the regular tumor vaccine. Therefore, the vaccines may become a novel potential therapy for recurrence and metastasis of HCC. PMID- 14562376 TI - Total vascular exclusion technique for resection of hepatocellular carcinoma. AB - AIM: To improve the low resection rate, poor prognosis and to control the massive hemorrhage during operation, total vascular exclusion (TVE) technique was used in hepatectomies of advanced and complicated hepatocellular carcinomas (HCCs). METHODS: Five hundred and thirty patients with HCCs were admitted in our hospital. They were divided into TVE technique group (group A: n=78), Pringle maneuver method group (group B: n=176) and unresectable group (group C: n=276). The clinical, operative, pathological parameters and outcome of the patients were statistically evaluated. RESULTS: Group A had a significantly higher resection rate than group B (accounting for 47.92% and 33.21% respectively). There was no significant difference in blood loss, blood transfusion and perioperative mortality between groups A and B. Both groups had the similar median disease free survival time (14.6 vs 16.3 months) and 1 year survival rate (92.9% vs 95.5%). The TVE group had a medial survival time of 40.5 months and its 5-year survival rate was 34.6%. CONCLUSION: As compared with Pringle maneuver method, the total vascular exclusion is a safe and effective technique to increase the total resection rate of advanced and complicated HCCs. PMID- 14562375 TI - Oxidative DNA damage in peripheral leukocytes and its association with expression and polymorphisms of hOGG1: a study of adolescents in a high risk region for hepatocellular carcinoma in China. AB - AIM: To study the oxidative DNA damage to adolescents of hepatocellular carcinoma (HCC) families in Guangxi Zhuang Autonomous Region, China. METHODS: Peripheral leukocytes' DNA 7, 8-dihydro-8-oxoguanine (8-oxoG) and repair enzyme hOGG1 were quantified by flow-cytometry. hOGG1-Cys326Ser single nucleotide polymorphism (SNP) was distinguished by polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) assay. RESULTS: There was a positive correlation between 8-oxoG and repair enzyme hOGG1 expression (P<0.001). HCC children (n=21) in Fusui county had a higher level of hOGG1 (P<0.01) and a lower level of 8-oxoG (P<0.05) than the controls (n=63) in Nanning city. Children in Nanning exposed to passive smoking had a higher hOGG1 expression (P<0.05) than the non-exposers. 8-oxoG and hOGG1 were negatively correlated with body mass index, while hOGG1 was positively correlated with age. There was a peak of 8-oxoG level nearby the 12 year point. Individuals with the hOGG1 326Ser allele had a significantly marginal higher concentration of leukocyte 8-oxoG level than hOGG1 326Cys allele. CONCLUSION: This is the first report using flow-cytometry to simultaneously quantify both the DNA oxidative damage and its repairing enzyme hOGG1. The results provide new insights towards a better understanding of the mechanisms of oxidative stress in a population highly susceptible to hepatocarcinogenesis. PMID- 14562377 TI - Role of multiphase scans by multirow-detector helical CT in detecting small hepatocellular carcinoma. AB - AIM: To evaluate the role of multiphasic scanning by multirow-detector helical CT (MDCT) in detecting small hypervascular hepatocellular carcinoma (SHCC). METHODS: Multiphasic scanning was carried out in 75 patients with SHCC with Marconi MX8000 CT scanner. The early arterial phase (EAP), late arterial phase (LAP) and the portal venous phase (PVP) scans were started at 21 s, 34 s and 85 s respectively. The mean difference of CT values between tumor and liver parenchyma for each scanning phase was measured, and the sensitivity of detection of SHCC in each of these phases and in the combined phase was calculated and statistically analyzed. RESULTS: The mean difference of CT values between tumor and liver parenchyma was significant in 71 lesions > or =1 cm in three phases (P<0.05). In 91 tumor foci, the detectability of SHCC was 45.1%, 83.5% and 92.3% in EAP, LAP and double arterial phases (DAP), respectively. The early arterial phase plus the portal venous phase and the double arterial phase plus the portal venous phase were 94.5%, 97.8%, respectively. Whereas the detectability in LAP plus PVP and in DAP plus PVP had no statistical difference. CONCLUSION: The utility of faster speed and thinner slice MDCT and multiphase scanning protocol can improve the detectability of hypervascular small hepatocellular carcinoma. Among which LAP is superior to EAP in depicting the lesions. PMID- 14562378 TI - Expressing patterns of p16 and CDK4 correlated to prognosis in colorectal carcinoma. AB - AIM: To describe the correlation between immunostaining patterns of p16 and CDK4 and prognosis in colorectal carcinoma. METHODS: Paraffin sections of 74 cases of colorectal carcinoma were analysed immunohistochemically for expression of p16 and CDK4 proteins. RESULTS: Most carcinomas showed stronger p16 and CDK4 immunostaining in the cytoplasm than the adenomas or the adjacent normal mucosa. Strong immunostaining of p16 was a predictor for better prognosis whereas strong cytoplasmic immunostaining of CDK4 was a predictor for poor prognosis. Both p16 and CDK4 immunostainings were correlated with histological grade or Dukes' stage. CONCLUSION: These results support the experimental evidence that interaction of expression of p16 and CDK4 may play an important role in the Rb/p16 pathway, and the expression patterns of CDK4 and p16 may be imperative in the development of colorectal carcinoma, thus becoming a new prognostic marker in colorectal cancer. PMID- 14562379 TI - Prognostic factors in 165 elderly colorectal cancer patients. AB - AIM: To analyse the prognostic factors in 165 colorectal patients aged > or =70. METHODS: One hundred and sixty-five elderly patients with colorectal cancer diagnosed by histology were entered into the retrospective study between 1994 and 2001. Patients were given optimal operation alone, chemotherapy after operation, or chemotherapy alone according to tumor stage, histology, physical strength, and co-morbid problems. Survival rate was calculated by Kaplan-Meier method, and compared with meaningful variances by Log-rank method. Prognostic factors were analyzed by Cox regression. RESULTS: The 1,2,3,4,5 year survival rate (all-cause mortality) was 87.76 %, 65.96%, 52.05%, 42.77 %, 40.51%, respectively. The mean survival time was 41.89+/-2.33 months (95% CI: 37.33-46.45 months), and the median survival time was 37 months. Univariate analysis showed that factors such as age, nodal metastasis, treatment method, Duke's stage, gross findings, kind of histology, and degree of differentiation had influences on the survival rate. Multivariate analysis showed that factors such as treatment method, Duke's stage, kind of histology and degree of differentiation were independent prognostic factors. CONCLUSION: This study suggests that the prognosis of elderly colorectal cancer patients is influenced by several factors. Most of elderly patients can endure surgery and/or chemotherapy, and have a long-time survival and good quality of life. PMID- 14562380 TI - Expression of ORF2 partial gene of hepatitis E virus in tomatoes and immunoactivity of expression products. AB - AIM: To transfer hepatitis E virus (HEV) ORF2 partial gene to tomato plants, to investigate its expression in transformants and the immunoactivity of expression products, and to explore the feasibility of developing a new type of plant derived HEV oral vaccine. METHODS: Plant binary expression vector p1301E2, carrying a fragment of HEV open reading frame-2 (named HEV-E2), was constructed by linking the fragment to a constitutive CaMV35s promoter and nos terminator, then directly introduced into Agrobacterium tumefaciens EHA105. With leaf-disc method, tomato plants medicated by EHA105 were transformed and hygromycin resistant plantlets were obtained in selective medium containing hygromycin. The presence and integration of foreign DNA in transgenic tomato genome were confirmed by Gus gene expression, PCR amplification and Southern dot blotting. The immunoactivity of recombinant protein extracted from transformed plants was examined by enzyme-linked immunosorbant assay (ELISA) using a monoclonal antibody specifically against HEV. ELISA was also used to estimate the recombinant protein content in leaves and fruits of the transformants. RESULTS: Seven positive lines of HEV-E2-transgenic tomato plants confirmed by PCR and Southern blotting were obtained and the immunoactivity of recombinant protein could be detected in extracts of transformants. The expression levels of recombinant protein were 61.22 ng/g fresh weight in fruits and 6.37-47.9 ng/g fresh weight in leaves of the transformants. CONCLUSION: HEV-E2 gene was correctly expressed in transgenic tomatoes and the recombinant antigen derived from them has normal immunoactivity. Transgenic tomatoes may hold a good promise for producing a new type of low-cost oral vaccine for hepatitis E virus. PMID- 14562381 TI - Lethiferous effects of a recombinant vector carrying thymidine kinase suicide gene on 2.2.15 cells via a self-modulating mechanism. AB - AIM: To determine the lethiferous effects of a recombinant vector carrying thymidine kinase (TK) suicide gene on 2.2.15 cells and the possible self modulating mechanism. METHODS: A self-modulated expressive plasmid pcDNA3-SCITK was constructed by inserting the fragments carrying hepatitis B virus antisense-S (HBV-anti-S) gene, hepatitis C virus core (HCV-C) gene, internal ribosome entry site (IRES) element of HCV and TK gene into the eukaryotic vector pcDNA3, in which the expression of TK suicide gene was controlled by the HBV S gene transcription. 2.2.15 cells that carry the full HBV genome and stably express series of HBV antigen were transfected with pcDNA3-SCITK or vector pcDNA3-SCI which was used as the mock plasmid. The HepG2 cells transfected with pcDNA3-SCITK were functioned as the negative control. All the transfected cells were incubated in DMEM medium supplemented with 10 microg/ml. of ganciclovir (GCV). The HBsAg levels in the supernatant of cell culture were detected by ELISA on the 1st, 3rd and 6th day post-transfection. Meanwhile, the morphology of transfected cells was recorded by the photograph and the survival cell ratio was assessed by the trypan blue exclusion test on the 6th day post-transfection. RESULTS: The structural accuracy of pcDNA3-SCITK was confirmed by restriction endonuclease digestion, PCR with specific primers and DNA sequencing. The HBsAg levels in the supernatant of transfected 2.2.15 cell culture were significantly decreased on the 6th day post transfection as compared with that of the mock control (P<0.05). The lethiferous effect of pcDNA3-SCITK expression on 2.2.15 cells was initially noted on the 3rd day after transfection and aggravated on the 6th day post transfection, in which the majority of transfected 2.2.15 cells were observed shrunken, round in shape and even dead. With assessment by the trypan blue exclusion test, the survival cell ratio on the 6th day post transfection was 95% in the negative control and only 11% in the experimental group. CONCLUSION: The results indicate that suicide gene expression of pcDNA3-SCITK can only respond to HBV-S gene transcription, which may be potentially useful in the treatment of HBV infection and its related liver malignancies. PMID- 14562382 TI - Association between HLA class II gene and susceptibility or resistance to chronic hepatitis B. AB - AIM: To investigate the association between the polymorphism of HLA-DRB1, -DQA1 and -DQB1 alleles and viral hepatitis B. METHODS: HLA-DRB1, -DQA1 and -DQB1 alleles in 54 patients with chronic hepatitis B, 30 patients with acute hepatitis B and 106 normal control subjects were analyzed by using the polymerase chain reaction/sequence specific primer (PCR/SSP) technique. RESULTS: The allele frequency of HLA-DRB1*0301 in the chronic hepatitis B group was markedly higher than that in the normal control group (17.31% vs 5.67%), there was a significant correlation between them (chi2=12.3068, Pc=0.0074, RR=4.15). The allele frequency of HLA-DQA1*0501 in the chronic hepatitis B group was significantly higher than that in the normal control group (25.96% vs 13.68%), there was a significant correlation between them (chi2=9.2002, Pc=0.0157, RR=2.87). The allele frequency of HLA-DQB1*0301 in the chronic hepatitis B group was notably higher than that in the normal control group (35.58% vs 18.87%), there was a significant correlation between them (chi2=15.5938, Pc=0.0075, RR=4.07). The allele frequency of HLA DRB1*1101/1104 in the chronic hepatitis B group was obviously lower than that in the normal control group (0.96% vs 13.33%), there was a significant correlation between them (chi2=11.9206, Pc=0.0145, RR=18.55). The allele frequency of HLA DQA1*0301 in the chronic hepatitis B group was remarkably lower than that in the normal control group (14.42% vs 30%), there was a significant correlation between them (chi2=8.7396, Pc=0.0167, RR=0.35). CONCLUSION: HLA-DRB1*0301, HLA-DQA1*0501 and HLA-DQB1*0301 are closely related with susceptibility to chronic hepatitis B, and HLA-DRB1*1101/1104 and HLA-DQA1*0301 are closely related with resistance to chronic hepatitis B. These findings suggest that host HLA class II gene is an important factor determining the outcome of HBV infection. PMID- 14562383 TI - No requirement of HCV 5'NCR for HCV-like particles assembly in insect cells. AB - AIM: To express all three HCV structural proteins in the presence or absence of HCV 5'NCR to investigate the requirement of 5'NCR for the assembly of HCV-like particles in insect cells. METHODS: HCV structural protein encoding sequences CE1E2 and 5'NCR-CE1E2 were amplified with PCR. Recombinant baculovirus were constructed with recombinant DNA techniques. HCV structural proteins expressed in insect cells were analyzed by immunofluorescence and SDS-PAGE. Immunoprecipitation experiment of insect cell lysates with anti-E2 monoclonal antibody (MAb) was carried out and the immunoprecipitated proteins were subjected to SDS-PAGE and immunoblotting with anti-C, anti-E2 MAbs and HCV positive serum. The virus-like particles in insect cells were visualized by electron microscopy (EM). The HCV-like particles were purified by sucrose gradient centrifugation and identified by EM and immune aggregation EM. RESULTS: The recombinant baculovirus reBV/CE1E2 containing HCV C, E1, E2 genes and reBV/CS containing the same structural protein genes plus 5'NCR were constructed. The insect cells infected with either reBV/CE1E2 or reBV/CS expressed HCV C, E1 and E2 proteins with a molecular weight of 20 kD, 35 kD and 66 kD respectively. The results of immunoprecipitation and the immunoblotting revealed the coimmunoprecipitation of C, E1, and E2 proteins, indicating the interaction of HCV structural proteins expressed in insect cells. Electron microscopy of insect cells infected with reBV/CE1E2 or reBV/CS demonstrated spherical particles (40 to 60 nm in diameter) similar to the HCV virions from sera or hepatic tissues of HCV infected humans. The HCV-like particles were partially purified by sucrose gradient centrifugation, and the purified VLPs showed immuno-reactivity with anti-HCV antibodies. CONCLUSION: HCV 5'NCR is not required for the assembly of HCV-like particles in insect cells, HCV core and envelope proteins are sufficient for viral particle formation. PMID- 14562384 TI - Interobserver variation in histopathological assessment of Helicobacter pylori gastritis. AB - AIM: Because the presence or absence of H pylori infection has important implications for therapeutic decisions based on histological assessment, the reproducibility of Sydney system is important. The study was designed to test the reproducibility of features of Helicobacter pylori gastritis, using the updated Sydney classification. METHODS: Gastric biopsies of 40 randomly selected cases of H pylori gastritis were scored semiquantitatively by three pathologists. Variables analysed included chronic inflammation, inflammatory activity, atrophy, intestinal metaplasia, H pylori, surface epithelial damage. Kappa values below 0.5 represented poor, those between 0.5 and 0.75 good and values over 0.75 excellent interobserver agreement. RESULTS: The best interobserver agreement (kappa=0.62) was present for intestinal metaplasia. The agreement was the poorest for evaluating atrophy (kappa=0.31). CONCLUSION: Although the results of this study were in accordance with some previous studies, an excellent agreement could not be reached for any features of H pylori gastritis. This low degree of concordance is assumed to be due to the personal evaluation differences in grading the features, the lack of standardized diagnostic criteria, and the ignorance to reach a consensus about the methods to be used in grading the features of H pylori gastritis before initiating the study. PMID- 14562385 TI - Gastric polypoid lesions: analysis of 150 endoscopic polypectomy specimens from 91 patients. AB - AIM: To analyze gastric polypoid lesions in our patient-population with respect to histopathologic features and demographic, clinical, and endoscopic characteristics of patients. METHODS: Clinical records and histopathologic reports of patients with gastric polypoid lesions were analyzed retrospectively. All lesions had been totally removed by either endoscopic polypectomy or hot biopsy forceps. The histopathologic slides were re-evaluated by the same histopathologist. RESULTS: One-hundred and fifty gastric polypoid lesions were identified in 91 patients. There were 53 (58%) women and 38 (42%) men with a median age of 53 (range, 31 to 82) years. The most frequent presenting symptom was dyspepsia that was observed in 35 (38.5%) patients. Symptoms were mostly related to various associated gastric abnormalities such as chronic gastritis or H pylori infection rather than polypoid lesion itself. Polypoid lesions were commonly located in the antrum followed by cardia. Out of 150 lesions, 80 (53%) had the largest dimensions less than or equal to 5 mm and only 7 were pedunculated. The frequencies of hyperplastic polyps, foveolar hyperplasia, and fundic gland polyps were 46%, 18%, and 14% respectively. We also detected gastritis varioliformis in 12 specimens, lymphoid follicles in 9, 4 adenomatous polyps in 4, polypoid lesions with edematous mucosa in 4, inflammatory polyps in 3, and carcinoid tumor in 1. Adenomatous changes were observed within two hyperplastic polyps and low grade dysplasia in one adenoma. Histopathologic evaluation of the surrounding gastric mucosa demonstrated chronic gastritis in 72 (79%) patients and H pylori infection in 45 (49%). CONCLUSION: Hyperplastic polyps are the most frequently encountered subtype of gastric polypoid lesions. They are usually associated with chronic gastritis or H pylori gastritis. Contrary to the previous belief, they may harbour adenomatous changes or dysplastic foci. Therefore, endoscopic polypectomy seems as a safe and fast procedure for both diagnosis and treatment of gastric polypoid lesions at the same session. In addition, edematous mucosa may appear misleadingly as a polypoid lesion in some instances and it can be ruled out only by histopathologic examination. PMID- 14562386 TI - Construction of expression systems for flaA and flaB genes of Helicobacter pylori and determination of immunoreactivity and antigenicity of recombinant proteins. AB - AIM: To clone flagellin genes A (flaA) and B (flaB) from a clinical strain of Helicobacter pylori (H pylori) and to construct prokaryotic expression systems of the genes and identify immunity of the fusion proteins. METHODS: The flaA and flaB genes from a clinical H pylori isolate Y06 were amplified by high fidelity PCR. The nucleotide sequences of target DNA amplification fragments from the two genes were sequenced after T-A cloning. The recombinant expression vector pET32a inserted with flaA and flaB genes was constructed, respectively. The expressions of FlaA and FlaB fusion proteins in E. coli BL21DE3 induced by isopropylthio-beta D-galactoside (IPTG) at different concentrations were examined by SDS-PAGE. Western blot using commercial antibodies against whole cell of H pylori and immunodiffusion assay using self-prepared rabbit antiserum against FlaA (rFlaA) or FlaB (rFlaB) recombinant proteins were applied to the determination of the fusion proteins immunity. ELISA was used to detect the antibodies against rFlaA and rFlaB in sera of 125 H pylori infected patients and to examine rFlaA and rFlaB expression in 98 clinical isolates of H pylori, respectively. RESULTS: In comparison with the reported corresponding sequences, the nucleotide sequence homologies of the cloned flaA and flaB genes were from 96.28-97.13% and 96.31 97.73%, and their putative amino acid sequence homologies were 99.61-99.80% and 99.41-100% for the two genes, respectively. The output of rFlaA and rFlaB expressed by pET32a-flaA-BL21DE3 and pET32a-flaB-BL21DE3 systems was as high as 40-50% of the total bacterial proteins. Both rFlaA and rFlaB were able to combine with the commercial antibodies against whole cell of H pylori and to induce rabbits to produce specific antibodies with the same 1:2 immunodiffusion titers after the animals were immunized with the two recombinant proteins. Ninety-eight and zero point 4 and 92.80% of the serum samples from 125 patients infected with H pylori were positive for rFlaA and rFlaB antibodies, respectively. One hundred percent and 98.98% of the 98 tested isolates of H pylori were detectable for rFlaA and rFlaB epitopes, respectively. CONCLUSION: Two prokaryotic expression systems with high efficiency of H pylori flaA and flaB genes were successfully established. The expressed rFlaA and rFlaB showed satisfactory immunoreactivity and antigenicity. High frequencies of FlaA and FlaB expression in different H pylori clinical strains and the general existence of specific antibodies against FlaA and FlaB in H pylori infected patients strongly indicate that FlaA and FlaB are excellent antigen candidates for developing H pylori vaccine. PMID- 14562388 TI - Distribution of cagG gene in Helicobacter pylori isolates from Chinese patients with different gastroduodenal diseases and its clinical and pathological significance. AB - AIM: To determine the distribution of cagG gene of Helicobacter pylori (H pylori) isolates cultured from patients with various digestive diseases and its relationship with gastroduodenal diseases. METHODS: cagG was amplified by polymerase chain reaction in 145 H pylori isolates cultured from patients with chronic gastritis (n=72), duodenal ulcer (n=48), gastric ulcer (n=17), or gastric and duodenal ulcer (n=8), and the relationship between cagG status and the grade of gastric mucosal inflammation was determined. RESULTS: cagG was present in 91.7% of the 145 H pylori isolates, with the rates were 90.3%, 93.8%, 88.2% and 100.0%, respectively, in those from patients with chronic gastritis, duodenal ulcer, gastric ulcer, and gastric and duodenal ulcer. There was no significant difference among the four groups (P>0.05). The average grade of gastric mucosal inflammation in the antrum and corpus was 1.819+/-0.325 and 1.768+/-0.312, respectively in cagG positive patients, whereas the average inflammation grade was 1.649+/-0.297, 1.598+/-0.278 respectively in cagG negative cases (P>0.05). CONCLUSION: cagG gene of H pylori was quite conservative, and most H pylori strains in Chinese patients were cagG positive. cagG status was not related to clinical outcome or the degree of gastric mucosal inflammation. Therefore, cagG can not be used as a single marker for discrimination of H pylori strains with respect to a specific digestive disease. PMID- 14562387 TI - Deletion of Helicobacter pylori vacuolating cytotoxin gene by introduction of directed mutagenesis. AB - AIM: To construct a vacA-knockout Helicobacter pylori mutant strain, whose only difference from the wild strain is its disrupted vacA gene. METHODS AND RESULTS: A clone containing kanamycin resistance gene used for homologous recombination was constructed in a directional cloning procedure into pBluescript II SK, and then transformed into vacA+ H pylori by electroporation. Colonies growing on the selective media containing kanamycin were harvested for chromosomal DNA extraction, and the allelic exchange was determined by polymerase chain reactions and sequencing. Loss of vacuolating activity of the vacA-knockout strain was confirmed by examining the gastric cells co-cultured with cell-free supernatants from H pylori wild strain or the mutant. CONCLUSION: We constructed a vacA knockout strain of H pylori through direct mutagenesis, which creates an important precondition for the future research on virulence comparison with gene expression analysis. PMID- 14562389 TI - Effect of oral epidermal growth factor on mucosal healing in rats with duodenal ulcer. AB - AIM: To investigate the effect of epidermal growth factor (EGF) on mucosal healing in rats with duodenal ulcer. METHODS: Male Sprague-Dawley rats were randomly divided into sham operation without EGF, sham operation with EGF, duodenal ulcer without EGF, or duodenal ulcer with EGF groups. Additionally, normal rats without operation served as the control group. Duodenal ulcer was induced in rats by 300 mL/L acetic acid. Rats with EGF were orally administered at a dose of 60 microg/kg/day in drinking water on the next day of operation (day 1). Healing of duodenal ulcer was detected by haematoxylin and eosin staining. Cell growth of damaged mucosa was determined by the contents of nucleic acids and proteins. The level of EGF in duodenal mucosa was measured by ELISA. RESULTS: The pathological results showed that duodenal ulcer rats with EGF improved mucosal healing compared with those without EGF after day 5. Duodenal ulcer rats with EGF significantly increased duodenal DNA content compared with those without EGF on day 15 (6.44+/-0.54 mg/g vs 1.45+/-0.52 mg/g mucosa, P<0.05). Duodenal RNA and protein contents did not differ between duodenal ulcer rats with and without EGF during the experimental period. Sham operation and duodenal ulcer rats with EGF significantly increased duodenal mucosal EGF content compared with those without EGF on day 5 (76.0+/-13.7 ng/g vs 35.7+/-12.9 ng/g mucosa in sham operation rats, and 68.3+/-10.9 ng/g vs 28.3+/-9.2 ng/g mucosa in duodenal ulcer rats, P<0.05). CONCLUSION: Oral EGF can promote mucosal healing of the rats with duodenal ulcer by stimulating mucosal proliferation accompanied by an increase in mucosal EGF content. PMID- 14562390 TI - Role of oxygen free radicals in patients with acute pancreatitis. AB - AIM: The generation of oxygen free radicals has been implicated in the pathogenesis of experimental pancreatitis. The aim of this study was to determine the role of oxygen free radicals in patients with acute pancreatitis. METHODS: The plasma levels of C-reactive protein (CRP), lipid peroxide (LPO), myeloperoxidase (MPO) and superoxide dismutase (SOD) were measured in 13 patients with acute pancreatitis and 14 healthy volunteers. RESULTS: Among the patients with acute pancreatitis, there were higher plasma levels of LPO and MPO and lower SOD activity in patients with severe pancreatitis than in those with mild pancreatitis. However, there was no significant difference in the serum marker of oxidative stress no matter what the etiology was. The LPO level was especially correlated with the concentration of serum CRP and CT severity index. CONCLUSION: The oxygen free radicals may be closely associated with inflammatory process and the severity of acute pancreatitis. Especially, the concentration of plasma LPO is a meaningful index for determining the severity of the disease. PMID- 14562391 TI - Parenteral versus early intrajejunal nutrition: effect on pancreatitic natural course, entero-hormones release and its efficacy on dogs with acute pancreatitis. AB - AIM: To evaluate the effect of early intrajejunal nutrition (EIN) on the natural course, entero-hormone secretion and its efficacy on dogs with acute pancreatitis. METHODS: An acute pancreatitis model was induced by injecting 1 ml/kg of combined solution (2.5% sodium taurocholate and 8,000-10,000 BAEE units trypsin/ml) into the pancreas via pancreatic duct. Fifteen dogs were divided into parenteral nutrition (PN) group and EIN group. Two groups were isonitrogenous and isocaloric. EIN was used at postoperative 24 h. Serum glucose, calcium, amylase and lysosomal enzymes were determined before and 1, 4, 7 d after acute pancreatitis was induced. All the dogs were injected 50 uCi 125I-BSA 4 h before sacrificed on the 7th day. The 125I -BSA index of the pancreas/muscle, pancreas/blood, and pancreas pathology score (PPS) were determined. The peripheral plasma cholecystokinin (CCK), secretin (SEC) and gastrin were measured by ELISA and RIA, and was quantitative analysis of pancreatic juice and amylase, pancreatolipase and HCO3-, Cl-, Na+ and K+ performed by an autochemical analyzer at 30, 60, 120 and 180 min after beginning PN or EIN on the first day. RESULTS: There was no difference between two groups in the contents of serum calcium, amylase and lysosomal enzymes, 125I-BSA index of pancreas/muscle and pancreas/blood and PPS. The contents of CCK and gastrin in EIN were higher than those in PN group at 60 and 120 min (P<0.05). The content of SEC post-infusion of nutrition solution was higher than that of pre-infusion of nutrition solution in both groups, and only at 60 min SEC in EIN group was higher than that in PN group. The content of gastrin in EIN was higher than that in PN group at 120 and 180 min (P<0.05). The changes of pancreatic juice, amylase, pancreatolipase and HCO3-, Cl-, Na+ and K+ between two groups did not reach significantly statistical difference (P>0.05). CONCLUSION: EIN does not stimulate entero-hormone and pancreatic juice secretion, and enzyme-protein synthesis and release. EIN has no effect on the natural course of acute pancreatitis. PMID- 14562392 TI - Protective effects of transplanted and mobilized bone marrow stem cells on mice with severe acute pancreatitis. AB - AIM: To evaluate the protective effects of transplanted and mobilized bone marrow stem cells (BMSCs) on mice with severe acute pancreatitis (SAP) and to probe into their possible mechanisms. METHODS: A mouse model of SAP induced by intraperitoneal injections of L-arginine was employed in the present study. Two hundred female Balb/c mice weighing 18-22 g were randomly assigned into 4 groups. Group A was the stem cell mobilized group treated by injection of granulocyte colony stimulating factor (G-CSF) into mice for 4 days at a dose of 40 microg.kg( 1).d(-1) before induction of SAP. Group B was the group of BMSCs transplantation, in which the mice were given the isolated BMSCs via the tail vein 4 days prior to induction of SAP. Group C served as the model control and only SAP was induced. The mice without induction of SAP in group D acted as the normal control. At the time of animal sacrifice at 24, 48 and 72 h after induction of SAP, blood samples were obtained and prepared to detect serum amylase, while the abdominal viscera were examined both grossly and microscopically for the observation of pathological changes. RESULTS: The mortality of mice in the model control, groups A and B was 34%, 8% and 10% respectively within 72 h after induction of SAP. The serum level of amylase in the model control was significantly increased at all time points after induction of SAP as compared with that of the normal control (P<0.05-0.01). When the mice were pretreated with BMSCs' transplantation or G-CSF injection, their serum level of amylase was significantly reduced at 48 h and 72 h after induction of SAP in comparison with that of the model control (P<0.05 0.01). In accordance with these observations, both gross and microscopic examinations revealed that the pathological changes of SAP in mice pretreated with BMSCs transplantation or G-CSF injection were considerably attenuated as compared with those in the model control at all observed time points. CONCLUSION: Both transplanted allogenic and mobilized autologous BMSCs can protect mouse pancreas from severe damage in the process of SAP. PMID- 14562393 TI - Biomechanical properties of ileum after systemic treatment with epithelial growth factor. AB - AIM: Systemic treatment with epidermal growth factor (EGF) leads to growth of all parts of the small intestine in normal functioning rats. In this study, we investigated the effect of this growth process on morphometric and biomechanical parameters of ileum. METHODS: Rats were treated with EGF (150 microg.kg(-1)day( 1))or placebo via osmotic minipumps for 2, 4, 7, and 14 days. A segment of ileum was removed. The morphology at no-load state and zero-stress state was measured and passive biomechanical properties were assessed using a biaxial test machine (combined inflation and axial stretching). RESULTS: The ileum weight increased after EGF administration. After 4 days' EGF treatment, the wall thickness was increased. Significantly smaller inner perimeters were seen in 4 day and 7 day EGF treatment groups. The opening angle and residual strain began to increase after 7 days' EGF treatment. Wall stiffness, evaluated from the stress-strain curves, showed a continuous decrease in circumferential direction during the first 7 days' EGF treatment. The longitudinal stiffness increased during the first 7 days. The stress-strain curves for both circumferential and longitudinal direction tended to shift back to normal 14 days after starting EGF administration. CONCLUSION: EGF can cause significant changes both in the morphology and in the passive mechanical properties of the rat ileum. PMID- 14562394 TI - Effects of Rheum tanguticum polysaccharide on TNBS -induced colitis and CD4+T cells in rats. AB - AIM: To study the effects of Rheum tanguticum polysaccharide(-1) (RTP(-1)) on ulcerative colitis in rats induced by 2, 4, 6-trinitrophene sulphonic acid (TNBS) and their possible mechanism. METHODS: RTP1 (200 mg.kg(-1), i.g.) extracted from Rheum tanguticum Maxim. ex Regel was administrated to rats with colitis induced by TNBS for 5 d, 7 d, 10 d and 14 d, respectively. The effects of RTP1 and dexamethasone (DX, 0.2 mg.kg(-1), i.g.) were contrastively investigated. The MPO level and SOD activity were determined by chromatometry. The expansion and protein expression of CD4+T lymphocytes isolated from colon mucosae and mesenteric lymph nodes of colitis rats were performed by immunohistochemical analysis and Western-blot methods. RESULTS: Treatments of RTP1 (200 mg.kg(-1), i.g.) significantly reduced diarrhea, mortality, colon mass, ulcer areas and MPO level in colon mucosae on days 5, 7, 10 and 14 (5.2+/-1.4, 5.4+/-0.7, 5.2+/-1.8, P<0.05. 3.4+/-0.8, P<0.01. 16.1+/-12.1, P<0.01. 31.8+/-8.6, 17.7+/-5.3, 12.7+/ 4.1, P<0.05). The effects of RTP1 were similar to those noted above in DX group, but there were no immunosuppressive effects of DX in RTP(-1) group, such as body mass loss, thymus and spleen atrophy. The decreased number and down-regulated protein levels of CD4+T cells isolated from the colon of colitis rats treated with RTP1 were found. CONCLUSION: RTP1 shows significantly protective effects but lower side effects on rats with colitis induced by TNBS. The mechanism may be due to the resistance to over expansion of CD4. PMID- 14562395 TI - Effect of ischemic preconditioning on P-selectin expression in hepatocytes of rats with cirrhotic ischemia-reperfusion injury. AB - AIM: To investigate the effects and mechanisms of ischemic preconditioning (IPC) on the ischemia/reperfusion (I/R) injury of liver cirrhosis in rats and the effect of IPC on P-selectin expression in hepatocytes. METHODS: Forty male SD rats with liver cirrhosis were randomly divided into sham operation group (SO group), ischemia/reperfusion group (I/R group), ischemic preconditioning group (IPC group), L-Arginine preconditioning group (APC group), L-NAME preconditioning group (NPC group), eight rats in each group. Hepatocellular viability was assessed by hepatic adenine nucleotide level and energy charge (EC) determined by HPLC, ALT, AST and LDH in serum measured by auto- biochemical analyzer and bile output. The expression of P-selectin in the liver tissue was analyzed by immunohistochemical technique. Leukocyte count in ischemic hepatic lobe was calculated. RESULTS: At 120 min after reperfusion, the level of ATP and EC in IPC and APC groups was higher than that in I/R group significantly. The increases in AST, ALT and LDH were prevented in IPC and APC groups. The livers produced more bile in IPC group than in I/R group during 120 min after reperfusion (0.101+/ 0.027 versus 0.066+/-0.027 ml/g liver, P=0.002). There was a significant difference between APC and I/R groups, (P=0.001). The leukocyte count in liver tissues significantly increased in I/R group as compared with SO group (P<0.05). The increase in the leukocyte count was prevented in IPC group. Administration of L-arginine resulted in the same effects as in IPC group. However, inhibition of NO synthesis (NPC group) held back the beneficial effects of preconditioning. Significant promotion of P-selectin expression in hepatocytes in the I/R group was observed compared with the SO group (P<0.01). IPC or L-arginine attenuated P selectin expression remarkably (P<0.01). However, inhibition of NO synthesis enhanced P-selectin expression (P<0.01). The degree of P-selectin expression was positively correlated with the leukocyte counts infiltrating in liver (r=0.602, P=0.000). CONCLUSION: IPC can attenuate the damage induced by I/R in cirrhotic liver and increase the ischemic tolerance of the rats with liver cirrhosis. IPC can abolish I/R induced leukocyte adhesion and infiltration by preventing post ischemic P-selectin expression in the rats with liver cirrhosis via a NO initiated pathway. PMID- 14562396 TI - Pan-enteric dysmotility, impaired quality of life and alexithymia in a large group of patients meeting ROME II criteria for irritable bowel syndrome. AB - AIM: Psychological factors, altered motility and sensation disorders of the intestine can be variably associated with irritable bowel syndrome (IBS). Such aspects have not been investigated simultaneously. The aim of this paper was to evaluate gastrointestinal motility and symptoms, psychological spectrum and quality of life in a large group of IBS patients in southern Italy. METHODS: One hundred IBS patients (F:M=73:27, age 48+/-2 years, mean+/-SE) fulfilling ROME II criteria matched with 100 healthy subjects (F:M=70:30, 45+/-2 years). Dyspepsia, bowel habit, alexithymia, psycho-affective profile and quality of life were assessed using specific questionnaires. Basally and postprandially, changes in gallbladder volumes and antral areas after liquid meal and orocaecal transit time (OCTT) were measured respectively by ultrasonography and H(2)-breath test. Appetite, satiety, fullness, nausea, and epigastric pain/discomfort were monitored using visual-analogue scales. RESULTS: Compared with controls, IBS patients had increased dyspepsia (score 12.6+/-0.7 vs 5.1+/-0.2, P<0.0001), weekly bowel movements (12.3+/-0.4 vs 5.5+/-0.2, P<0.00001, comparable stool shape), alexithymia (score 59.1+/-1.1 vs 40.5+/-1.0, P=0.001), poor quality of life and psycho-affective profile. IBS patients had normal gallbladder emptying, but delayed gastric emptying (T50: 35.5+/-1.0 vs 26.1+/-0.6 min, P=0.00001) and OCTT (163.0+/-5.4 vs 96.6+/-1.8 min, P=0.00001). Fullness, nausea, and epigastric pain/discomfort were greater in IBS than in controls. CONCLUSION: ROME II IBS patients have a pan-enteric dysmotility with frequent dyspepsia, associated with psychological morbidity and greatly impaired quality of life. The presence of alexithymia, a stable trait, is a novel finding of potential interest to detect subgroups of IBS patients with different patterns recovered after therapy. PMID- 14562398 TI - Seroreactivity against Saccharomyces cerevisiae in patients with Crohn's disease and celiac disease. AB - AIM: To explore whether there was anti-Saccharomyces cerevisiae antibodies (ASCA) positivity in our patients with biopsy-confirmed celiac disease. METHODS: A cohort of patients with inflammatory bowel diseases (42 patients with Crohn's disease and 10 patients with ulcerative colitis) and gluten sensitive enteropathy (16 patients) from Debrecen, Hungary were enrolled in the study. The diagnosis was made using the formally accepted criteria. Perinuclear antineutrophil cytoplasmic antibodies (pANCA) and anti-Saccharomyces cerevisiae antibodies (ASCA), antiendomysium antibodies (EMA), antigliadin antibodies (AGA) and anti human tissue transglutaminase antibodies (tTGA) were investigated. RESULTS: The results showed that ASCA positivity occurred not only in Crohn's disease but also in Celiac disease and in these cases both the IgG and IgA type antibodies were proved. CONCLUSION: It is conceivable that ASCA positivity correlates with the (auto-) immune inflammation of small intestines and it is a specific marker of Crohn's disease. PMID- 14562397 TI - Association of extraintestinal manifestations of inflammatory bowel disease in a province of western Hungary with disease phenotype: results of a 25-year follow up study. AB - AIM: IBD is a systemic disease associated with a large number of extraintestinal manifestations (EIMs). Our aim was to determine the prevalence of EIMs in a large IBD cohort in Veszprem Province in a 25-year follow-up study. METHODS: Eight hundred and seventy-three IBD patients were enrolled (ulcerative colitis/UC/: 619, m/f: 317/302, mean age at presentation: 38.3 years, average disease duration: 11.2 years; Crohn's disease/CD/: 254, m/f: 125/129, mean age at presentation: 32.5 years, average disease duration: 9.2 years). Intestinal, extraintestinal signs and laboratory tests were monitored regularly. Any alteration suggesting an EIMs was investigated by a specialist. RESULTS: A total of 21.3% of patients with IBD had EIM (UC: 15.0%, CD: 36.6%). Age at presentation did not affect the likelihood of EIM. Prevalence of EIMs was higher in women and in CD, ocular complications and primary sclerosing cholangitis (PSC) were more frequent in UC. In UC there was an increased tendency of EIM in patients with a more extensive disease. Joint complications were more frequent in CD (22.4% vs UC 10.2%, P<0.01). In UC positive family history increased the risk of joint complications (OR:3.63). In CD the frequency of type-1 peripheral arthritis was increased in patients with penetrating disease (P=0.028). PSC was present in 1.6% in UC and 0.8% in CD. Dermatological complications were present in 3.8% in UC and 10.2% in CD, the rate of ocular complications was around 3% in both diseases. Rare complications were glomerulonephritis, autoimmune hemolytic anaemia and celiac disease. CONCLUSION: Prevalence of EIM in Hungarian IBD patients is in concordance with data from Western countries. The high number of EIM supports a role for complex follow-up in these patients. PMID- 14562399 TI - Clinical utility, safety and tolerability of capsule endoscopy in urban Southeast Asian population. AB - AIM: Capsule endoscopy has demonstrated its clinical utility in the evaluation of small bowel pathology in several Western studies. In this prospective study, we aimed to determine the clinical utility, safety and tolerability of capsule endoscopy in the evaluation of suspected small bowel disease in an urban Southeast Asian population. METHODS: We used the given (M2A) capsule endoscopy system in 16 consecutive patients with suspected small bowel pathology. In 9 patients the indication was obscure gastrointestinal bleeding, while in 6 patients it was to determine the extent of small bowel involvement in Crohn's disease. One patient underwent capsule endoscopy for evaluation of chronic abdominal pain. Patient's tolerability to the procedure was evaluated by standardized questionnaires and all patients were reviewed at one week to ensure that the capsule had been excreted without any adverse events. RESULTS: Abnormal findings were present in 8 patients (50%). The cause of obscure gastrointestinal bleeding was determined in 5 out of 9 patients. Findings included 2 cases of angiodysplasia, 2 cases of jejunal ulcers and 1 case of both angiodysplasia and jejunal ulcer. One patient had small bowel erosions and foci of erythema of doubtful significance. Ileal lesions were diagnosed in 2 out of 6 patients with Crohn's disease. Capsule endoscopy was well tolerated by all patients. One patient with Crohn's disease had a complication of capsule retention due to terminal ileum stricture. The capsule eventually passed out spontaneously after 1 month. CONCLUSION: Our study, which represented the first Asian series, further confirms the diagnostic utility, safety and tolerability of wireless capsule endoscopy. PMID- 14562401 TI - Healthy ranges of serum alanine aminotransferase levels in Iranian blood donors. AB - AIM: The healthy ranges for serum alanine aminotransferase (ALT) levels are less well studied. The aim of this study was to define the upper limit of normal (ULN) for serum ALT levels, and to assess factors associated with serum ALT activity in apparently healthy blood donors. METHODS: A total of 1,939 blood donors were included. ALT measurements were performed for all cases using the same laboratory method. Healthy ranges for ALT levels were computed from the population at the lowest risk for liver disease. Univariate and multivariate analyses were performed to evaluate associations between clinical factors and ALT levels. RESULTS: Serum ALT activity was independently associated with body mass index (BMI) and male gender, but not associated with age. Association of ALT with BMI was more prominent in males than in females. Upper limit of normal for non overweight women (BMI of less than 25) was 34 U/L, and for non-overweight men was 40 U/L. CONCLUSION: Serum ALT is strongly associated with sex and BMI. The normal range of ALT should be defined for male and female separately. PMID- 14562400 TI - Budd-Chiari syndrome: diagnosis with three-dimensional contrast-enhanced magnetic resonance angiography. AB - AIM: To evaluate the role of three-dimensional contrast-enhanced magnetic resonance angiography (3D CE MRA) in the diagnosis of Budd-Chiari syndrome (BCS). METHODS: Twenty-three patients with BCS underwent 3D CE MRA examination, in which 13 cases were secondary to either hepatocellular carcinoma (11 cases), right adrenal carcinoma (1 case) or thrombophlebitis (1 case) and 10 suffered from primary BCS. The patency of the inferior vena cava (IVC), hepatic and portal veins as well as the presence of intra- and extrahepatic collaterals, liver parenchymal abnormalities and porto-systemic varices were evaluated. Inferior vena cavography was performed in 10 cases. The diagnosis of IVC obstruction by 3D CE MRA was compared with that demonstrated by inferior vena cavography. RESULTS: The major features of BCS could be clearly displayed on 3D CE MRA. Positive hepatic venous signs included tumor thrombosis (9 cases), tumor compression (2 cases), nonvisualization (4 cases) and focal stenosis (2 cases). Positive IVC findings were noted as severe stenosis or occlusion (10 cases), tumor invasion (2 cases), thrombosis (3 cases), thrombophlebitis (1 case) and septum formation (3 cases). Intrahepatic collaterals were shown in 9 patients, 2 of them with "spider web" sign. The displayed extrahepatic collaterals included dilated azygos and hemi-azygos veins (13 cases) and left renal-inferior phrenic-pericardiophrenic veins (2 cases). The occlusion of the left intrahepatic portal veins was found in 2 cases. Porto-systemic varices were detected in 10 patients. Liver parenchymal abnormalities displayed by 3D CE MRA were enlargement of the caudate lobe (7 cases), heterogenous enhancement (18 cases) and complicated tumors (13 cases). Compared with the inferior vena cavography performed in 10 cases, the accuracy of 3D CE MRA was 100 % in the diagnosis of IVC obstruction. CONCLUSION: 3D CE MRA can display the major features of BCS and provide an accurate diagnosis. PMID- 14562402 TI - Adrenomedullin in cirrhotic and non-cirrhotic portal hypertension. AB - AIM: Adrenomedullin (ADM) is a potent vasodilator peptide. ADM and nitric oxide (NO) are produced in vascular endothelial cells. Increased ADM level has been linked to hyperdynamic circulation and arterial vasodilatation in cirrhotic portal hypertension (CPH). The role of ADM in non-cirrhotic portal hypertension (NCPH) is unknown. plasma ADM levels were studied in patients with NCPH, compensated and decompensated cirrhosis in order to determine its contribution to portal hypertension (PH) in these groups. METHODS: There were 4 groups of subjects. Group 1 consisted of 27 patients (F/M: 12/15) with NCPH due to portal and/or splenic vein thrombosis (mean age: 41+/-12 years), group 2 consisted of 14 patients (F/M: 6/8) with compensated (Child-Pugh A) cirrhosis (mean age: 46+/-4), group 3 consisted of 16 patients (F/M: 6/10) with decompensated (Child-Pugh C) cirrhosis (mean age: 47+/-12). Fourteen healthy subjects (F/M: 6/8) (mean age: 44+/-8) were used as controls in Group 4. ADM level was measured by ELISA. NO was determined as nitrite/nitrate level by chemoluminescence. RESULTS: ADM level in Group 1 (236+/-61.4 pg/mL) was significantly higher than that in group 2 (108.4+/ 28.3 pg/mL) and group 4 (84.1+/-31.5 pg/mL) (both P<0.0001) but was lower than that in Group 3 (324+/-93.7 pg/mL) (P=0.002). NO level in group 1 (27+/-1.4 micromol/L) was significantly higher than that in group 2 (19.8+/-2.8 micromol/L) and group 4 (16.9+/-1.6 micromol/L) but was lower than that in Group 3 (39+/-3.6 micromol/L) (for all three P<0.0001). A strong correlation was observed between ADM and NO levels (r=0.827, P<0.0001). CONCLUSION: Adrenomedullin and NO levels were high in both non-cirrhotic and cirrhotic portal hypertension and were closely correlated, Adrenomedullin and NO levels increased proportionally with the severity of cirrhosis, and were significantly higher than those in patients with NCPH. Portal hypertension plays an important role in the increase of ADM and NO. Parenchymal damage in cirrhosis may contribute to the increase in these parameters. PMID- 14562403 TI - Candida esophagitis: risk factors in non-HIV population in Pakistan. AB - AIM: Candida esophagitis is a frequent infection in immunocompromised patients. This study was designed to determine its characteristics in non- human immune deficiency virus (HIV) infected patients attending a teaching hospital. METHODS: Clinical records of all patients coded by international classification of diseases 9th revision with clinical modifications' (ICD-9-CM), with candida esophagitis diagnosed by esophagogastroduodenoscopy (EGD) and histopathology over a period of 5 years were studied. RESULTS: Fifty-one patients (27 males, 24 females, range 21-77 years old and mean age 52.9 years) fulfilled the criteria (0.34% of the EGD). The common predisposing factors were carcinoma (OR 3.87, CI 1.00-14.99) and diabetes mellitus (OR 4.39, CI 1.34-14.42). The frequent clinical symptoms were retrosternal discomfort, dysphagia and epigastric abdominal pain with endoscopic appearance of scattered mucosal plaques. Another endoscopic lesion was associated with candida esophagitis in 15% patients. CONCLUSION: Carcinomas, diabetes mellitus, corticosteroid and antibiotic therapy are major risk factors for candida esophagitis in Pakistan. It is an easily managed complication that responds to treatment with nystatin. PMID- 14562404 TI - Serum malondialdehyde level in patients infected with Ascaris lumbricoides. AB - AIM: The aim of the study was to investigate the changes of serum malondialdehyde level, i.e; the oxidative stress hypothesis in patients infected with Ascaris lumbricoides. METHODS: Serum malondialdehyde activity was measured in 43 patients who were positive for intestinal parasite of Ascaris lumbricoides. Scores were obtained for the positives and their age-and sex-matched 60 Ascaris lumbricoides negative healthy controls. RESULTS: The difference between malondialdehyde levels of patients infected with Ascaris lumbricoides and control group was statistically significant both for females (P<0.05) and for males (P<0.05). In the patient and control groups, no correlation was found between age and malondialdehyde levels (P>0.05) both in females and in males. In addition, no significant correlation could be found between malondialdehyde levels of both females and males for patients and control groups (P>0.05). CONCLUSION: Malondialdehyde levels clearly increase in patients infected with Ascaris lumbricoides. PMID- 14562405 TI - Problems in screening colorectal cancer in the elderly. AB - AIM: To explore the problems in the screening of colorectal carcinoma in the elderly. METHODS: Three models of colorectal cancer prevention were examined: standard screening, active check-up of suspected cases and summons to have endoscopic check-up for previously diagnosed colorectal polyps. The study was performed among three groups of elderly individuals: Group 1 (167 cases), hospitalized asymptomatic individuals without symptoms in large intestines. Group 2 (612 cases): old individuals at home for the aged, out of which 32 showed symptoms of colon disorders; Group 3 (44 cases): elderly people with diagnosed polyps. As a result of 1788 rectosigmoidoscopies, we identified 61 individuals with polyps, out of which 44 patients were over 65 years old. However, only 9 of these 44 individuals agreed to have the endoscopy performed again. RESULTS: One cancer and 13 polyps were detected in Group 1, and two polyps in Group 2. However, it should be noted that only eleven individuals from Group 2 agreed to have the endoscopy. In Group 3, there were no relapses of the polyps among the nine individuals who came back for the endoscopy. CONCLUSION: Poor understanding of the screening procedures is one of the greatest problems in early detection of the cancer in the aged. Paradoxically, the cooperation is better with hospitalized patients, than with "successfully old" persons. PMID- 14562406 TI - Significant factors associated with fatal outcome in emergency open surgery for perforated peptic ulcer. AB - AIM: To evaluate the main factors associated with mortality in patients undergoing surgery for perforated peptic ulcer referred to an academic department of general surgery in a large southern Italian city. METHODS: One hundred and forty-nine consecutive patients (M:F ratio=110:39, mean age 52 yrs, range 16-95) with peptic ulcer disease were investigated for clinical history (including age, sex, previous history of peptic ulcer, associated diseases, delayed abdominal surgery, ulcer site, operation type, shock on admission, postoperative general complications, and intra-abdominal and/or wound infections), serum analyses and radiological findings. RESULTS: The overall mortality rate was 4.0%. Among all factors, an age above 65 years, one or more associated diseases, delayed abdominal surgery, shock on admission, postoperative abdominal complications and/or wound infections, were significantly associated (chi2) with increased mortality in patients undergoing surgery (0.00010.05). CONCLUSION: When induced during cultivation in vitro, PBL-Ts from CHB have AICD very commonly. This phenomenon has a potentially important relation with pathogenesis of CHB and chronicity of HBV infection. PMID- 14562413 TI - Temporary partially-covered metal stent insertion in benign esophageal stricture. AB - AIM: To study the therapeutic efficacy of temporary partially-covered metal stent insertion on benign esophageal stricture. METHODS: Temporary partially-covered metal stent was inserted in 83 patients with benign esophageal stricture. All the patients had various dysphagia scores. RESULTS: Insertion of 85 temporary partially-covered metal stents was performed successfully in 83 patients with benign esophageal stricture and dysphagia was effectively remitted in all the 83 cases. The dysphagia score was 3.20+/-0.63 (mean+/-SD) and 0.68+/-0.31 before and after stent insertion, and 0.86+/-0.48 after stent removal. The mean diameter of the strictured esophageal lumen was 3.37+/-1.23 mm and 25.77+/-3.89 mm before and after stent insertion, and 16.15+/-2.96 mm after stent removal. Follow-up time was from 1 week to 96 months (mean 54.26+/-12.75 months). The complications were chest pain (n=37) after stent insertion, and bleeding (n=12) and reflux (n=13) after stent removal. CONCLUSION: Temporary partially-covered metal stent insertion is one of the best methods for treatment of benign esophageal stricture. PMID- 14562414 TI - Antiproliferative effect of octreotide on gastric cancer cells mediated by inhibition of Akt/PKB and telomerase. AB - AIM: To investigate the antiproliferative effect of octreotide, a long-acting analogue of somatostatin, on gastric cancer cell line SGC7901 and its possible molecular mechanisms. METHODS: Gastric cancer cell line SGC7901 employed in the study was treated with 0.008, 0.04, 0.2, 1, 5 and 25 microg.ml(-1) of octreotide respectively for 24 h to evaluate the antiproliferative effect of somatostatin analog on the tumor cells by MTT assay method. To elucidate the underlying mechanism, the cells were exposed to 1 microg.ml(-1) of octreotide for 0, 12, 24 and 48 h, when their Akt/PKB and telomerase activities were respectively determined using PCR-ELSIA and nonradioactive protein kinase assay protocols. The same experimental procedures were also performed in the control cells that were treated with corresponding vehicles instead of somatostatin analog. RESULTS: After exposed to octreotide for 24 h at the concentrations of more than 1 microg.ml(-1), SGC7901 cells exhibited a dose-dependent inhibition of growth with the inhibiting rate to be as high as 34.66% when 25 microg.ml(-1) of octreotide was applied. The Akt/PKB and telomerase activity of SGC7901 cells was significantly inhibited when the cells were exposed to 1 microg.ml(-1) of octreotide for 12, 24 and 48 h compared with that of their control counterparts (P<0.01), both of which exhibited in a time-dependent manner. CONCLUSION: The antiproliferative effect of octreotide on SGC7901 cells might be mediated by the inhibition of Akt/PKB and telomerase. PMID- 14562416 TI - Selection and evaluation of three interventional procedures for achalasia based on long-term follow-up. AB - AIM: To determine the best method out of the three types of interventional procedure for achalasia based on a long-term follow-up. METHODS: The study cohort was comprised of 133 patients of achalasia. Among them, 60 patients were treated under fluoroscopy with pneumatic dilation (group A), 8 patients with permanent uncovered or antireflux covered metal stent dilation (group B), and 65 patients with temporary partially covered metal stent dilation (group C). RESULTS: One hundred and thirty dilations were performed on the 60 patients of group A (mean 2.2 times per case). The mean diameter of the strictured cardia was 3.3+/-2.1 mm before dilation and 10.6+/-3.8 mm after dilation. The mean dysphagia score was 2.7+/-1.4 before dilation and 0.9+/-0.3 after dilation. Complications in group A were chest pain (n=30), reflux (n=16), and bleeding (n=6). Thirty-six patients (60%) in group A exhibited dysphagia relapse during a 12-month follow-up, and 45 patients (90%) out of 50 exhibited dysphagia relapse during a 36-month follow-up. Five uncovered and 3 antireflux covered expandable metal stents were permanently placed in the 8 patients of group B. The mean diameter of the strictured cardia was 3.4+/-1.9 mm before dilation and 19.5+/-1.1 mm after dilation. The mean dysphagia score was 2.6+/-1.3 before dilation and 0.4+/-0.1 after dilation. Complications in group B were chest pain (n=6), reflux (n=5), bleeding (n=3), and hyperplasia of granulation tissue (n=3). Four patients (50%) in group B exhibited dysphagia relapse during a 12-month follow-up, and 2 case (66.7%) out of 3 patients exhibited dysphagia relapse during a 36-month follow-up. Sixty-five partially covered expandable metal stents were temporarily placed in the 65 patients of group C and withdrawn after 3-7 days via gastroscopy. The mean diameter of the strictured cardia was 3.3+/-2.3 mm before dilation and 18.9+/-3.5 mm after dilation. The mean dysphagia score was 2.4+/-1.3 before dilation and 0.5+/-0.2 after dilation. Complications in group C were chest pain (n=26), reflux (n=13), and bleeding (n=8). 6 patients (9.2%) out of 65 exhibited dysphagia relapse during a 12-month follow-up, and 8 patients (14.5%) out of 55 exhibited dysphagia relapse during a 36-month follow-up. All the stents were inserted and withdrawn successfully. The follow-up in groups A-C lasted 12-96 months. CONCLUSION: Temporary partially covered metal stent dilation is one of the best methods with interventional procedure for achalasia in terms of long-term follow up. PMID- 14562417 TI - Effects of glycine on plasma and liver tissue changes of TNF-alpha, ET-1 and nitric oxide contents in rats with obstructive jaundice. AB - AIM: To evaluate the effect of glycine on plasma and liver tissue changes of tumor necrosis factor-alpha (TNF-alpha), endothelin-1 (ET-1) and nitric oxide (NO) contents in rats with obstructive jaundice. METHODS: Ninety healthy Wistar rats of both sexes weighing 275+/-25 g were randomly divided into sham-operated, bile duct-ligated, and bile duct-ligated plus glycine-treated groups, the latter was performed with 5% glycine solution substituting for tap water drunk ad libium for 5 days before and 6 days after operation. Blood and liver tissue were sampled at the time of sacrifice on the 8th day post operation. Plasma total bilirubin, endotoxin, levels, as well as TNF-alpha, ET-1 and NO contents in liver tissue were determined. RESULTS: Plasma endotoxin and total bilirubin levels were significantly higher in both bile duct-ligated and bile duct-ligated plus glycine treated rats than in sham-operated animals (P=0.000613, 0.00921 and 0.00737, 0.00841 respectively), whereas they did not display any statistically significant difference between the former groups (P=0.417 and 0.374 respectively). Likewise, TNF-alpha, ET-1 and NO contents in both plasma and liver tissue were significantly increased in both bile duct-ligated and bile duct-ligated plus glycine-treated rats compared with sham-operated animals(P=0.00813, 0.00793, 0.00671, 0.00804, 0.00872, and 0.00947 in plasma and 0.00531, 0.00785, 0.00912, 0.00981 and 0.00635 in liver tissue respectively). However, these inflammatory mediators in both plasma and liver tissue were significantly reduced in bile duct ligated rats fed on 5% glycine solution compared with that without (P=0.00953, 0.00891, 0.0795, 0.00867, 0.0697 and 0.00907 in plasma and liver tissue respectively). CONCLUSION: Reduction of TNF-alpha, ET-1 and NO contents in plasma and liver tissue of rats fed on glycine may be helpful to alleviate pathological lesions in obstructive jaundice. PMID- 14562415 TI - Molecule action mechanisms of NM-3 on human gastric cancer SGC-7901 cells in vivo or in vitro. AB - AIM: To study the molecule action mechanisms of NM-3 on the growth of human gastric cancer SGC-7901 cells in vivo or in vitro. METHODS: SGC-7901 from human non-differentiated gastric cancer cell line was cultured with NM-3 at 100 mg/ml for 24 h. We observed its inhibitory rate and the density of micro-vascular growth in grafted mice with human gastric cancer SGC-7901. The apoptosis of human gastric cancer SGC-7901 was revealed in NM-3 treatment group by using terminal deoxynucleotidyl transferase-mediated deoxy-uridine triphosphate-fluorescene nick end labeling (TUNEL) method and flow cytometry analysis. RESULTS: The growth of SGC-7901 cells was markedly inhibited compared with control growp, which was smaller than that in normal saline control group (4.17 g+/-0.22 g vs 9.45 g+/ 1.38 g, P<0.01). The level of apoptosis of human gastric cell line SGC-7901 was obviously increased in NM-3 treatment group at 1 mg.L(-1) for 24 h. NM-3 inducing apoptotic index in NM-3 plus carboplatin group was 3.5 times that of carboplatin control group (TUNEL: 27.98+/-6.12% vs 12.94+/-2.12%, FACScan: 26.86+/-5.69% vs 11.86+/-1.09%, P<0.01). Western blot analysis showed that the apoptotic index of human gastric cancer was elevated for 12, 24 and 36 h with an evident time-effect relationship in groups at 100 mg.L(-1). NM-3 enhanced the inhibitive effects and sensitivity of chemotherapy for human gastric cancer in nude mice. These results suggested that NM-3 played a key inhibitive role in the growth of grafted human gastric cancer in nude mice. CONCLUSION: NM-3 can inhibit the growth of human gastric cancer cell line SGC-7901, and enhance the sensitivity of carboplatin on SGC-7901 and induced its apoptosis. PMID- 14562418 TI - Potentially fatal haemobilia due to inappropriate use of an expanding biliary stent. AB - AIM: To highlight the fatal complication caused by expanding biliary stents and the importance of avoiding use of expanding stent in potentially curable diseases. METHODS: Arteriobiliary fistula is an uncommon cause of haemobilia. We describe a case of right hepatic artery pseudoaneurysm causing arteriobiliary fistula and presenting as severe malena and cholangitis, in a patient with a mesh metal biliary stent. The patient had lymphoma causing bile duct obstruction. RESULTS: Gastroduodenoscopy failed to establish the exact source of bleeding and hepatic artery angiography and selective embolisation of the pseudo aneurysm successfully controlled the bleeding. CONCLUSION: Bleeding from the pseudo aneurysm of the hepatic artery can be fatal. Mesh metal stents in biliary tree can cause this complication as demonstrated in this case. So mesh metal stent insertion should be avoided in potentially benign or in curable conditions. Difficulty in diagnosis and management is discussed along with the review of the literature. PMID- 14562420 TI - Massive gastrointestinal tuberculosis in a young patient without immunosuppression. AB - Although the lung is the major site for Mycobacterium tuberculosis infection, gastrointestinal involvement can be present as part of multiorgan disease process or, less commonly, can be seen as primary gastrointestinal tuberculosis. In the cases where the culture is negative, it can be difficult to differentiate tuberculosis from Crohn's disease based on both the clinical and histological features. When side effects of classic antimycobacteria are encountered, we can initially add ciprofloxacin to the treatment of tuberculosis. We reported a case of 19-yr-old patient, who was treated as Crohn's disease and worsen. We began to tuberculosis treatment, and the patient improved clinically and histologically. The main point in this case is that widespread involvement of gastrointestinal tract can be brought about by non resistant strains of Mycobacterium tuberculosis even in immunocompetent patients. PMID- 14562419 TI - Liver cell adenoma with malignant transformation: a case report. AB - A 57-year-old woman was referred to our hospital because of a liver mass detected by computed tomography. She had taken oral contraceptives for only one month at the age of thirty. Physical examination revealed no abnormalities, and laboratory data, including hepatic function tests, were within the normal range, with the exception of elevated levels of those serum proteins induced by the absence of vitamin K or by raised levels of the antagonist (PIVKA)-II (3,502 AU/ml). Abdominal ultrasonography revealed a hyperechoic mass measuring 10 X 10 cm in the left posterior segment of the liver. Because hepatocellular carcinoma could not be completely excluded, this mass was resected. The tumor consisted of sheets of uniform cells with clear cytoplasm, perinuclear eosinophilic granules and round nuclei. These histological findings were consistent with liver cell adenoma. Background hepatic tissue appeared normal. After resection of the tumor, serum PIVKA-II fell to within the normal range. An area of hepatocellular carcinoma (HCC) with a mid-trabecular pattern was immunohistochemically found, which was positive for PIVKA-II. Sinusoidal endothelial cells were CD34-positive, containing scattered PIVKA-II positive cells. This tumor was therefore finally diagnosed as liver cell adenoma with focal malignant transformation to HCC. PMID- 14562421 TI - Advocates for a healthy community. PMID- 14562422 TI - Child abuse and neglect reporting in Hawaii: the role of dentists. PMID- 14562423 TI - Comprehensive approach to the management and prevention of early childhood caries. AB - Dental caries is the most common disease of childhood that does not resolve spontaneously or respond to a course of antibiotic therapy. Tragically, children in Hawaii experience the highest incidence of tooth decay in the United States. This manuscript briefly explains the etiology of Early Childhood Caries (ECC), describes the consequences of this insidious disease process and outlines a comprehensive, multidisciplinary, preventive approach for Hawaii's health care providers. PMID- 14562424 TI - The future of dentistry in Hawaii. PMID- 14562425 TI - Regulating the practice of dentistry in Hawaii. PMID- 14562426 TI - Access to dental care in Hawaii. PMID- 14562427 TI - Are the NMC's fees value for money? PMID- 14562428 TI - Changing attitudes in dementia care and the role of nurses. AB - Dementia care has been seen as a low priority, but with an estimated 700,000 people in the UK with dementia and an ageing population this is changing. The Alzheimer's Society will be holding events this week to raise awareness of dementia, sending a strong message to policy makers that dementia is a serious health issue. Nurses have also become active in fighting the old degenerative model of dementia and promoting the concept of person-centred care. PMID- 14562429 TI - What you need to know about ... diphtheria. PMID- 14562430 TI - Criteria for the safe discharge of patients from the recovery room. AB - Guidelines need to be in place to help nurses in the recovery room make appropriate and safe decisions when discharging patients to a surgical ward. Consciousness level, respiration, circulation, pain control, homeostasis and wound care should all be considered. Criteria from the Freeman Hospital provides practical guidance. PMID- 14562431 TI - A nurse-led service for tunnelled central venous catheter insertion. AB - The procedure of inserting tunnelled central venous catheters was once carried out by surgeons or radiologists at Glasgow NHS Trust. However, increased workloads meant that these health care professionals were left with less time to devote to these procedures, which resulted in unacceptable waiting times for those requiring them. After visiting several centres in England where nurses were already successfully inserting tunnelled central venous catheters with impressive audit results, we decided to set up a similar service. Following a successful bid for funding we now have a nurse-led service for the insertion of tunnelled central venous catheters with minimal waiting times and low complication rates. PMID- 14562432 TI - The management of dyspepsia. AB - Dyspepsia is a general term that describes pain or discomfort that is centred in the upper abdomen. It reportedly affects up to 40 per cent of adults in any one year. A test and treat strategy is now recommended for all patients with uncomplicated dyspepsia. Any patient presenting with alarm features, such as recurrent vomiting or dysphagia, should be referred to a specialist for further investigation. PMID- 14562433 TI - Sharing medication agreements with patients. PMID- 14562434 TI - 'I feel like we've really cracked it'. PMID- 14562435 TI - Equip us to care. Buying power. PMID- 14562436 TI - Molecular spin clusters: new synthetic approaches and neutron scattering studies. AB - We review our recent work in the field of molecular spin clusters and single molecule magnets, showing how inelastic neutron scattering (INS) can be used to determine magnetic exchange interactions and anisotropy splittings. A general introduction to neutron scattering precedes selected examples, building upon the first determination of exchange coupling in a transition metal complex using INS, through anisotropic exchange in cobalt(II) spin clusters to the determination of exchange interactions in a dodecanuclear nickel(II) wheel. The strength of INS for the accurate determination of anisotropy splittings in single-molecule magnets is revealed. Not only can one determine the axial zero-field splitting parameter D, which plays a key role in single-molecule magnet behavior, but also higher-order terms important in understanding the quantum tunneling behavior. Finally, we review two of our synthetic approaches towards new single-molecule magnets based on nickel, manganese, and iron. PMID- 14562437 TI - Atomic resolution of oxygen in solid oxides by electron microscopy: what next? PMID- 14562438 TI - New insights into the Jahn-Teller effect through ab initio quantum mechanical/molecular-mechanical molecular dynamics simulations of CuII in water. AB - The CuII hydration shell structure has been studied by means of classical molecular dynamics (MD) simulations including three-body corrections and hybrid quantum-mechanical/molecular-mechanical (QM/MM) molecular dynamics (MD) simulations at the Hartree-Fock level. The copper(II) ion is found to be six-fold coordinated and [Cu(H2O)6]2+ exhibits a distorted octahedral structure. The QM/MM MD approach reproduces correctly the experimentally observed Jahn-Teller effect but exhibits faster inversions (< 200 fs) and a more complex behaviour than expected from experimental data. The dynamic Jahn-Teller effect causes the high lability of [Cu(H2O)6]2+ with a ligand-exchange rate constant some orders or magnitude higher than its neighbouring ions NiII and ZnII. Nevertheless, no first shell water exchange occurred during a 30-ps simulation. The structure of the hydrated ion is discussed in terms of radial distribution functions, coordination numbers, and various angular distributions and the dynamical properties as librational and vibrational motions and reorientational times were evaluated, which lead to detailed information about the first hydration shell. Second-shell water-exchange processes could be observed within the simulation time scale and yielded a mean ligand residence time of approximelty 20 ps. PMID- 14562439 TI - Ionic liquid for in situ Vis/NIR and Raman spectroelectrochemistry: Doping of carbon nanostructures. AB - 1-butyl-3-methylimidazolium tetrafluoroborate (an ionic liquid) is an advantageous electrolyte for the study of charge-transfer reactions at single walled carbon nanotubes (SWCNTs) and fullerene peapods (C60@SWCNT). Compared to traditional electrolyte solutions, this medium offers a broader window of electrochemical potentials to be applied, and favorable optical properties for in situ Vis/NIR and Raman spectroelectrochemistry of nano-carbon species. The electrochemistry of both nanotubes and peapods is dominated by their capacitive double-layer charging. Vis/NIR spectroelectrochemistry confirms the charging induced bleaching of transitions between Van Hove singularities. At high positive potentials, new optical transitions were activated in partly filled valence band. The bleaching of optical transitions is mirrored by the quenching of resonance Raman scattering in the region of tube-related modes. The Raman frequency of the tangential displacement mode of SWCNT shifts to blue upon both anodic and cathodic charging in the ionic liquid. The Raman modes of intratubular C60 exhibit a considerable intensity increase upon anodic doping of peapods. PMID- 14562440 TI - Time-resolved spectroscopic study on photoinduced electron-transfer processes in Zn(II)porphyrin-Zn(II)chlorin-fullerene triad. AB - Femtosecond time-resolved transient absorption studies have been performed to investigate the photoinduced energy and electron-transfer processes in Zn(II)porphyrin-Zn(II)chlorin-fullerene triad in which energy and oxidation potential gradients are directed along the donor-acceptor-linked arrays. Fast energy transfer (approximately 450 fs) from photoexcited Zn(II)porphyrin to Zn(II)chlorin was observed upon selective photoexcitation of Zn(II)porphyrin unit in the triad. In a nonpolar solvent such as toluene, the energy transfer from the excited singlet state of Zn(II)chlorin to fullerene occurs and is followed by the formation of an intermediate state with a time constant of nanoseconds, which was attributed to the intramolecular exciplex between Zn(II)chlorin and fullerene. In benzonitrile, on the other hand, the photoexcitation of the triad results in the fast electron transfer (< 1 ps) from photoexcited Zn(II)chlorin to fullerene. The generated charge-separated species recombine with a time constant of approximately 12 ps. The relatively fast charge separation and charge recombination rates imply that the strong electronic coupling between Zn(II)chlorin and fullerene moieties is probably induced by the folded conformation between Zn(II)chlorin and fullerene moieties which enhances direct through-space interaction between the approximately contacted pi systems. PMID- 14562441 TI - Long-range transport in an assembly of ZnO quantum dots: the effects of quantum confinement, Coulomb repulsion and structural disorder. AB - We have studied the storage and long-range transport of electrons in a porous assembly of weakly coupled ZnO quantum dots permeated with an aqueous and a propylene carbonate electrolyte solution. The number of electrons per ZnO quantum dot is controlled by the electrochemical potential of the assembly; the charge of the electrons is compensated by ions present in the pores. We show with optical and electrical measurements that the injected electrons occupy the S, P, and D type conduction electron levels of the quantum dots; electron storage in surface states is not important. With this method of three-dimensional charge compensation, up to ten electrons per quantum-dot can be stored if the assembly is permeated with an aqueous electrolyte. The screening of the electron charge is less effective in the case of an assembly permeated with a propylene carbonate electrolyte solution. Long-range electron transport is studied with a transistor set-up. In the case of ZnO assemblies permeated with an aqueous electrolyte, two quantum regimes are observed corresponding to multiple tunnelling between the S orbitals (at a low occupation) and P orbitals (at a higher occupation). In a ZnO quantum-dot assembly permeated with a propylene carbonate electrolyte solution, there is a strong overlap between these two regimes. PMID- 14562442 TI - Crosslinked copolyimide membranes for phenol recovery from process water by pervaporation. AB - The effectiveness of different copolyimide membranes in the process of recovering phenol from water by pervaporation has been investigated. The polyimides were obtained by the polycondensation of 6FDA (4,4'-hexafluoro-isopropylidene diphthalic anhydride) with different diamines. The diamines 4 MPD (2,3,5,6 tetramethyl-1,4-phenylene diamine), 6FpDA (4,4'-hexafluoro-isopropylidene dianiline), 6FpODA (4,4'-bis-(4'-aminophenoxyphenyl)-hexafluoropropane), and DABA (3,5-diaminobenzoic acid) as a monomer providing a crosslinkable group, were used. In order to reach chemical stability at high phenol concentrations, the polymer structures were crosslinked with 1,10-decanediol and OFHD (2,2,3,3,4,4,5,5-octafluorohexanediol). Pervaporation experiments were performed at 60 degrees C, covering a concentration range of phenol between 2 and 11 wt.%. The best separation characteristics were obtained with a 6FDA-6FpDA/DABA 2:1 membrane crosslinked with 1,10-decanediol. Using a 7.8 wt.% phenol feed mixture, a total flux of 14 kg microns m-2 h-1 was reached with an enrichment of 40 wt.% phenol in the permeate. It was found that conditioning the membrane using high phenol concentrations (between 8 and 11 wt.%) is a necessary pretreatment in order to enhance the flux and improve enrichment, especially if process water with low phenol concentrations is to be treated. In addition to the experimental results, a comparison with rubbery membrane materials is presented in the discussion. PMID- 14562443 TI - Electrochemical studies of vitamin K1 microdroplets: electrocatalytic hydrogen evolution. AB - The voltammetry of a basal-plane pyrolytic graphite electrode modified with a random ensemble of unsupported microdroplets of vitamin K, is investigated when the electrode is immersed in aqueous electrolytes. It is shown that in dilute acidic solutions, electroreduction occurs in a single two-electron two-proton process to yield the corresponding hydroquinone at the electrode?vitamin K1 microdroplet?aqueous-electrolyte three-phase boundary. On addition of ionic alkali-metal salts to the aqueous acidic phase, the electrochemical reduction of vitamin K1 to the quinol is accompanied by catalytic hydrogen evolution within and alkali-metal-cation insertion into the organic microdroplets. In strongly alkaline solutions, electrochemical reduction of vitamin K1 at the triple-phase junction is proposed as being a single two-electron process with concomitant uptake of alkali-metal cations in order to maintain electroneutrality within the oil phase. Surprisingly, the relative ease of cation insertion into the oil phase is demonstrated to be governed by the degree of ion-pair formation rather than by the Gibbs transfer energy of the cation across the liquid?liquid interface. PMID- 14562444 TI - Electroluminescent diodes from complementary discotic benzoperylenes. PMID- 14562445 TI - Probing solid surfaces with single polymers. PMID- 14562446 TI - The one-photon and two-photon absorption properties for porphyrin-derived monomers and dimers. PMID- 14562447 TI - On the covalency of silver-fluorine bonds in compounds of silver(I), silver(II) and silver(III). PMID- 14562448 TI - Polarization-dependent surface-enhanced Raman spectroscopy of isolated silver nanoaggregates. PMID- 14562449 TI - Maximum-likelihood approach to single-molecule polarization modulation analysis. PMID- 14562450 TI - Gas-phase reactivity of uracil, 2-thiouracil, 4-thiouracil, and 2,4-dithiouracil towards the Cu+ cation: a DFT study. PMID- 14562451 TI - Tunable surface oxidation states in Si/SiO2 nanostructures prepared from Si/SiQ2 mixtures and phenol hydroxylation activity. PMID- 14562452 TI - Local formation of an alloy by atomic contact between the STM tip and the substrate surface. PMID- 14562453 TI - The impact of domestic violence on the maternal-child relationship and preschool age children's functioning. AB - Although preschool-age children are at risk for witnessing domestic violence, the majority of research has focused on children ages 6 to 12 years. This study examines the mediating role of the mother-child relationship on preschool-age children's functioning in families experiencing domestic violence. Maternal report and behavioral observations of mother-child interactions were used to assess relationship quality. Participants consisted of 103 children and their mothers. The data fit the model well but indicated that some of the correlations were not in the expected direction. Although depressed mothers were struggling with parenting, other mothers appeared to be compensating for the violence by becoming more effective parents. Domestic violence negatively impacted children's behavior with their mothers in interactions but did not influence maternal report of problem behaviors, suggesting that the impact of domestic violence begins very early and in the realm of relationships rather than in mental health. PMID- 14562454 TI - The intergenerational transmission of relationship violence. AB - This study explored the intergenerational transmission of violence in a community sample. A telephone survey of 1,249 adults in the City of Vancouver assessed family-of-origin violence (father to mother, mother to father, father to self, and mother to self), as well as physical and psychological abuse in intimate relationships. All forms of family-of-origin violence were predictive of all forms of relationship abuse, consistent with a general social learning model of relationship violence. There was no evidence of gender-specific or role-specific patterns of transmission. For example, father-to-mother violence was not specifically predictive of men's perpetration and women's victimization in adult relationships. Nor was parent-to-self violence more predictive of victimization than perpetration. The methodological and theoretical implications of these findings are discussed. PMID- 14562455 TI - Are life stressors associated with marital violence? AB - The current study examined the link between different conceptualizations of life stressors and physical violence against spouses. Life stressors were measured in several different ways to test whether stressor frequencies and perceived impacts, life domains of stressors (i.e., loss, threat), and the nature of stressors (i.e., occupational, interpersonal) are correlates of men's and women's moderate and severe violence. Also explored were potential mediators and moderators of the stress-violence relationship. Community and clinic couples participated in this study. Results indicated that occupational and loss stressors were associated with men's violence whereas a wider array of stressors were associated with women's violence. In addition, stressors only discriminated between violent and nonviolent men whereas some stressors also discriminated between moderately and severely violent women. Depressive symptoms moderated the association between stressor impact and violence such that impact and women's violence were significantly correlated for women with elevated depressive symptoms. Results are discussed in light of theoretical and clinical implications. PMID- 14562456 TI - Children's coping with marital conflict and their adjustment and physical health: vulnerability and protective functions. AB - Children's strategies for coping with parental marital conflict were examined as predictors, mediators, and moderators of the relations between marital conflict and 8- to 11-year-olds' internalizing, externalizing, and physical health problems. In the context of marital conflict, a higher level of active coping and support coping combined was a protective factor against girls' depression symptoms and self-esteem problems and both boys' and girls' health problems. Further, avoidance coping was a vulnerability factor for externalizing, internalizing, and physical health problems in boys, and distraction coping was protective against children's depression and health problems. These findings extend the literature by delineating coping strategies that either protected children against, or heightened their vulnerability to, adjustment and health problems associated with exposure to parental marital conflict. PMID- 14562457 TI - Categories and continua of destructive and constructive marital conflict tactics from the perspective of U.S. and Welsh children. AB - Categories and continua of parents' marital conflict tactics based on multiple, conceptually grounded criteria were tested. Participants were 175 U.S. children, ages 8-16 years (88 boys, 87 girls) and 327 Welsh children, ages 11-12 years (159 boys, 168 girls). Children's responses (affective, cognitive, behavioral) to analog presentations of 10 everyday marital conflict tactics enacted by fathers or mothers showed substantial variation as a function of tactic used. Orderings of conflict tactics on the various response criteria varied as a function of moderators, particularly the gender of the parent expressing the conflict tactic. Conflict tactics were classified as either constructive or destructive according to criteria derived from the emotional security hypothesis. Except for calm discussion, classifications did not change regardless of cultural group, parent gender, or child age or gender. Recommendations for negotiating everyday marital conflict for the children's sake are discussed. PMID- 14562458 TI - Siblings' differential experiences of marital conflict and differences in psychological adjustment. AB - The present study examined whether siblings experience marital conflict differently and whether such differences, if present, were associated with differences in their adjustment. Self-report data about marital conflict, children's depressed mood, behavioral conduct, and externalizing problems were obtained from 122 sibling pairs (mean ages = 10 and 12 years) and their parents. Results indicated that siblings were significantly different in exposure to and appraisals of marital conflict. Differences in siblings' exposure to marital conflict were significantly correlated with differences in their depressed mood, behavioral conduct, and externalizing problems. Differences in siblings' feelings of self-blame for marital conflict were significantly correlated with differences in their depressed mood and behavioral conduct. Children who experienced more marital conflict than their siblings had more adjustment problems than their siblings. These results highlight the importance of studying siblings' unique experiences of marital conflict to better understand its impact on children's adjustment. PMID- 14562460 TI - Parents of 5-year-old in vitro fertilization children: psychological adjustment, parenting stress, and the influence of subsequent in vitro fertilization treatment. AB - Sixty-six parents who had conceived by in vitro fertilization (IVF) were compared with 46 matched naturally conceiving control parents regarding psychosocial adjustment and parenting stress 5 years after the birth of their first child. IVF mothers reported a more external locus of control than did control mothers but did not differ on other measures. Within the IVF group, higher levels of treatment predicted lower parenting stress and more defensive responding on the Parenting Stress Index (R. Abidin, 1990). The more positive reporting of high treatment IVF parents may be attributable to either defensive responding or the fact that they are particularly highly motivated and competent parents. The study findings confirm a growing body of research regarding overall positive adjustment in IVF parents but also highlight the importance of considering individual differences among IVF mothers with respect to treatment experience. PMID- 14562459 TI - The development of attitudes about physical punishment: an 8-year longitudinal study. AB - We examined young adolescents' endorsement of parental use of corporal punishment to elucidate processes underlying the intergenerational transmission of discipline strategies. The community sample was ethnically and socioeconomically diverse. Mothers completed interviews and questionnaires when the target children were entering kindergarten (n = 566) and in 6th and 8th grades. Adolescents completed questionnaires when they were in 8th grade (n = 425). Adolescents' attitudes about corporal punishment varied widely. Those adolescents who had been spanked by their own mothers were more approving of this discipline method, regardless of the overall frequency, timing, or chronicity of physical discipline they had received. However, there was no correlation among adolescents for whom physical maltreatment in early or middle childhood was suspected. PMID- 14562461 TI - Adolescent mothers' relationship with their children's biological fathers: social support, social strain, and relationship continuity. AB - Two-hundred and eighteen low-income, minority, adolescent mothers were interviewed during the perinatal period and 3 years later about their social networks, including their relationships with their children's fathers. Few adolescents were involved with fathers at both time points. Relationships with fathers were, in general, less supportive and less problematic over time. Moreover, although father support was not associated with adolescent mothers' psychological adjustment, father absence and father strain had negative associations with psychological adjustment. Maternal grandmother support buffered the negative effects of strain in the adolescents' relationships with biological fathers. Perceptions of less social support from maternal grandmothers and more social support from fathers during the perinatal period as well as less social support from a new male partner at 3 years postpartum predicted relationship continuity between adolescent mothers and fathers at 3 years postpartum. Implications for intervention and policy are discussed. PMID- 14562462 TI - Mothers' differential treatment of their adolescent childbearing and nonchildbearing children: contrasts between and within families. AB - This study examined mothers' differential treatment of adolescent sibling pairs in 2 family contexts: families with and without an adolescent childbearing daughter. Results based on mothers' ratings and children's ratings revealed that the mothers of childbearing daughters treated all of their children less affectionately than did the mothers of nonchildbearing adolescents. In families with a childbearing daughter, mothers expected a brighter future for and treated their never-pregnant daughter(s) more favorably than their childbearing daughter, and mothers' harsh treatment toward their children was correlated with high financial stress, excessive time spent caring for their daughter's child, and younger children's sexual behavior and drug and alcohol use. Findings highlight the preferential parenting that occurs within the families of childbearing teens and suggest its ramifications. PMID- 14562463 TI - Work-parenting linkages among dual-earner couples at the transition to parenthood. AB - This study evaluates the impact of work experiences on parenting quality during the transition to parenthood. Dual-earner parents (n = 83) completed measures of work experiences (autonomy and interpersonal atmosphere). Parenting was observed twice, 3 months apart. Hierarchical regression analyses showed that mothers who reported a more negative interpersonal atmosphere at work showed decreases in positive parenting and increases in negative parenting over time, after controlling for their occupational status and general well-being. Mothers' work experiences also predicted changes in fathers' observed parenting. In contrast, fathers' work experiences were largely unrelated to fathers' or mothers' parenting. With one exception, parents' occupational status did not moderate work parenting relationships, and feelings of role overload did not mediate these relationships. PMID- 14562464 TI - Social support and the quality of parenting under economic pressure and workload in Finland: the role of family structure and parental gender. AB - This study focused on how factors outside the home affect the quality of mothering and fathering. Economic pressure and workload were evaluated along with the compensating role of social support on parenting. Information was gathered from 842 mothers and 573 fathers including 139 single-mother and 21 single-father families. The results showed that the nature of the strains, together with parental gender and family structure, influenced their effects on parenting. The results further revealed some gender- and strain-specific protective functions of social support on parenting. For example, economic pressure was related to increased punitive parenting, which was compensated by instrumental and emotional support among the mothers. Workload was related to less authoritative single fathering, which was compensated by instrumental support. PMID- 14562465 TI - Conflict, social support, and relationship quality: an observational study of heterosexual, gay male, and lesbian couples' communication. AB - Data from 42 heterosexual, 46 gay male, and 33 lesbian couples were used to assess the contribution of conflict and support discussions to relationship quality. Couples completed questionnaires, and videotaped discussions were coded for levels of negative and positive behaviors. Correlations showed that behaviors were associated with relationship quality in the expected directions. Hierarchical linear modeling analyses assessed the unique contributions of individual and dyadic behaviors to the variability of relationship quality. The findings indicated that, beyond the contribution of individual negative behaviors in the conflict task, the variables of dyadic positive behaviors in the conflict task, individual positive behaviors in the support task, and perceived help accounted for unexplained variance in relationship quality. There were no differences between types of couples on levels of behaviors or on their contributions to relationship quality. PMID- 14562466 TI - Sex variations in the disclosure to parents of same-sex attractions. AB - The decision whether to disclose same-sex attractions to parents was explored through in-depth interviews with 164 young women and men. Participants were more likely to disclose to mothers than fathers, usually around age 19 years and in a face-to-face encounter. Mothers were told before fathers, largely because mothers asked or because youth wanted to share their life with them; fathers were told by someone other than their child or by the youth because it was time. The reason participants did not disclose to mothers was because it was not the right developmental time; the reason they did not disclose to fathers was because they were not close to them. Sons, more than daughters, feared the negative reactions of parents, who generally reacted in the same manner--supportive or slightly negative. Relationships with parents since disclosure generally had not changed or had improved. Sex of parent mattered more than sex of child on most domains. PMID- 14562467 TI - [Gen-ethics]. AB - Knowledge is always positive, but its application can not be so. The practice with Knowledges and techniques that can affect essential aspects of the beings must be guided by perfectly established rules. The World Committee on Bioethics elaborate the Universal Declaration on the Human Genome and Human Rights in November of 1997 with the support of all the countries of the world, with no exception, and of all the scientific communities it was established that human cloning can not be used with reproductive purposes. It is very important to make an adequate use of the terminology. Scientists must use a precise language and some groups must be careful before making a pronouncement about specific subjects outside their preparation. PMID- 14562468 TI - [Regulation of reproductive medicine in North America or the Wild West of Medicine (Part I)]. AB - The objective of this work is to offer a general introduction of the legal regulation of the Reproductive Medicine that is practiced in the United States of America. In the United States, besides the Federal Administration, there are also fifty state juridical instances who have competence in Assisted Reproduction subjects. The first section studies the limits of the Right to Reproduction. The American Constitution, interpreted by the Supreme Court, has been useful to establish the limits. Then, will be analyzed the reasons that have been taken by the legislator to adopt a permission with regard to issues appeared by the Medicine of the Fertility. Once studied the general context of the regulation of the Procreation Technology, it is exposed the state legislation and jurisprudence about the matter, and it is also made a review to the most relevant doctrine opinions of the american jurist in relation to Assistance to Reproduction. The cultural and juridical answers that these new clinic methods have used to control, improve or limit the reproduction capacity of human beings are almost as interesting as the Reproductive Technology. Contrary to what happens in Europe, where the legislator has firmly establish the regulation of the Reproductive Medicine since the late 80's, in North-America the lack of regulation has caused a free market where all kind of treatments of fertility are offered, just up to the point that an American author has called the sector "wild west of the Medicine". PMID- 14562469 TI - Embryonic stem cell research: can the law balance ethical, scientific and economic values? (Part I). PMID- 14562470 TI - [Bioethics in Latin America: legal perspective]. AB - The jurist's work is to detect the legal guiding principles, analize them and to anticipate what kind of acceptance they will have. The legislator must be prudent if the subject studied is changeable as it happens with the norms applied in Bioethics. This detection process is more delicate if the guiding principles that have to be detected are valid for such an extensive region, as it is Latin America, where the legislation of the different countries that form it would adopt them. The two problems that will be studied here are: a) if it is advisable or not to raise some Bioethic basic principles to the constitutional level. b) which are the main principles that have been adopted by the juridical legislations of Latin America and who, in some way, guide the legal regulation. PMID- 14562471 TI - [Right to life vs right to a determined quality of life. Reflections on human cloning]. AB - This article studies the bioethic divergences appeared as a consequence of the use and therapeutic obtaining of stem cells. It also chronicles the development of the cloning techniques and then, following the parameters of Bioethics, it distinguishes a categorization of the cloning with reproductive aims and cloning with therapeutic aims. It emphasizes how this problematic takes us to the beginning of human life and to the determination of the moment where this one deserves ethic recognition and legal protection. It also marks that the impact that would have the development of the therapeutic-cloning over the quality of life of sick persons and old men changes the traditional points of view. It is necessary a urgent normative with intenational validity, nevertheless the real safeguard against abuses would only be possible by the respect to human dignity by all the people who are involve in the investigation of these techniques. PMID- 14562472 TI - [Bioartificiality thresholds: oscillations of genetic patenting]. AB - The legal interpretations that both patent offices and courts in the US and in Europe have applied to biotechnological inventions seem to go back an forth in defining to what extent DNA sequences can be patented. This process has given rise to the creation of different thresholds of bioartificiality, and to the blurring of boundaries between what can and cannot be patented. The different legal rules established in both systems during the last decade have been also oscillating between increased international harmonization and concern for the internal regulatory framework. PMID- 14562473 TI - [Revocation of authorizations on GMO under the precautionary principle (with special attention to the Justice Tribunal of the European Communities]. AB - During the last decade, the european institutions have done a great effort in order to introduce the precautionary principle, applicable to the management of serious and uncertain risks. Under the protection of the precautionary principle, european authorities and authorities of the member states have partially modified, suspended or revoked authorizations relative to genetically modified organisms. Is it rightful this use of the precautionary principle? This is the matter that the European Communities Justice Court has faced when the affected companies have lodge their appeals against this decisions. PMID- 14562474 TI - Does new biotechnology and medicine need another type of bioethical input or is it an ethical conflict of interest? PMID- 14562475 TI - [Human oocyte freezing in Spain: comment on the Real Decree 120/2003]. AB - This work analyzes some scientific, ethic and legal aspects about the freezing of human ovocites in Spain. Inside the context of the Law 35/1998 of Assisted Reproduction Techniques, it is analyzed the Royal Decree 120/2003 which regulates the authorization of controlled experiences with reproductive aims. These experiences investigate the fecundation of ovocites or ovaric tissue previously frozen. It is also studied the inform made through parliament by the General Court of Judicial Power and it is also valued the importance of the terms "experience", "investigation" and "project" used in the legal text. PMID- 14562476 TI - Chinese people's attitudes towards genetic diseases and children with handicaps. PMID- 14562477 TI - Report on research with stem cells. PMID- 14562478 TI - [Real Decree 120/2003, of January 31, regulating the requirements for controlled studies, with reproductive purposes, of fertilization of previously frozen oocyte or ovarian tissue, related to assisted human reproduction techniques]. PMID- 14562479 TI - [Law 9/2003, of April 25, establishing the legal procedure of confined utilization, voluntary liberation, and commercialization of genetically modified organisms]. PMID- 14562480 TI - APN transitional care. PMID- 14562481 TI - APN transitional care. PMID- 14562482 TI - Nursing and the media. PMID- 14562483 TI - Nursing and the media. PMID- 14562484 TI - Author Barroso responds to letter. PMID- 14562485 TI - Adherence to long-term therapies: evidence for action. PMID- 14562486 TI - Nurses fighting against severe acute respiratory syndrome (SARS) in Hong Kong. PMID- 14562487 TI - Media portrayal of nurses' perspectives and concerns in the SARS crisis in Toronto. AB - PURPOSE: To describe nursing work life issues as portrayed in the media during the SARS crisis in Toronto. METHODS: Content analysis of local and national news media documents in Canada. Media articles were sorted and classified by topic, and themes were identified. FINDINGS: Themes were: (a) changing schemas of nursing practice: the new normal; (b) barriers to relational nursing work; (c) work life concerns: retention and recruitment; (d) nursing virtue: nurses as heroes and professionals; (e) paradoxical responses to nurses from the community; and (f) leadership in nursing during the SARS crisis. CONCLUSIONS: This analysis enhanced understanding of how nurses are portrayed in the media, but it indicated the significance of quality of work life and issues about work-home life. Some descriptions of the care and caring of nurses have made nursing seem like an important and influential profession to potential applicants who might previously have dismissed nursing as a career. PMID- 14562488 TI - Effects of coresidence and caregiving on health of Thai parents of adult children with AIDS. AB - PURPOSE: To explore potential effects on the health of older parents living with and caring for people infected with HIV. DESIGN: Comparison of health outcomes between affected parents and matched nonaffected parents, and between principal caregivers and nonprincipal caregivers in Thailand. METHODS: Survey data from 394 affected households and 376 nonaffected households; qualitative data from 18 interviews of affected older parents. RESULTS: A large proportion of older people with HIV-infected children provided time-consuming and strenuous caregiving services to them. Mothers shouldered most of this burden. Mothers who had a child die from AIDS reported lower levels of overall happiness than did mothers who had not. Mothers and fathers of PHAs (persons with HIV/AIDS) who died reported lower levels of overall happiness compared to 3 years previously (before the time of the death of their child) and compared to parents from households that did not experience an adult child's death. Many parents of children with AIDS experienced anxiety, insomnia, fatigue, muscle strain, and head and stomach aches during the time they cared for their ill children. CONCLUSIONS: Many older people suffered adverse health outcomes related to living with and caring for their children with AIDS. Recommendations indicate a variety of programs that might help such older parents. PMID- 14562489 TI - Increasing perceptions of self-worth in preadolescents diagnosed with ADHD. AB - PURPOSE: To test the effectiveness of a school-based, nurse-facilitated support group in increasing perceptions of scholastic competence, social acceptance, behavioral conduct, perceived athletic competence, perceived physical appearance, and perceived global self-worth in preadolescents diagnosed with attention deficit disorder (ADD) or attention deficit hyperactivity disorder (ADHD). DESIGN: A quasi-experimental design with 65 preadolescents diagnosed with ADD or ADHD in an upper-middle class community in the United States. METHODS: Participants randomly assigned to either the control or experimental group completed Harter's Self-Perception Profile for Children instrument at the beginning of the study and 4 weeks later. Students in the experimental condition participated in a school-nurse facilitated support group which met twice weekly for 4 weeks. FINDINGS: Participants in the support group had increased scores on each of the six subscales, with significant increases on four of the subscales, including perceived social acceptance, perceived athletic competence, perceived physical appearance, and perceived global self-worth. CONCLUSION: Participation in a school-based, nurse-led support group was positively associated with perception of self-worth in preadolescents diagnosed with ADD or ADHD. PMID- 14562490 TI - Sexually abstinent African American adolescent females' descriptions of abstinence. AB - PURPOSE: To describe sexual abstinence from the perspective of abstinent African American female adolescents. DESIGN: Descriptive qualitative. METHODS: Data were collected from 14 sexually abstinent, African American adolescent girls during two semi-structured interviews. Data were collected using the life history method and were analyzed in the style of narrative analysis. FINDINGS: This analysis indicated four themes in descriptions of abstinence: limited information, categorization of sexual behaviors, activities that lead to intercourse, and abstinence as a self-determined choice. For most participants, "having sex" referred to heterosexual vaginal intercourse and being abstinent meant that one chose to refrain from intercourse. CONCLUSIONS: The participants' limited definition of abstinence might lead them to unknowingly put themselves at risk by engaging in other forms of genital sexual activities. They may think that they are protected because they believe that they are practicing abstinence. PMID- 14562491 TI - The effects of an educational intervention on antipsychotic-induced weight gain. AB - PURPOSE: To assess the effect of an educational intervention on antipsychotic induced weight gain among patients with schizophrenia. DESIGN AND METHODS: Quasi experimental. Seventy patients with a DSM-IV diagnosis of schizophrenia or schizoaffective disorder entered this 6-month study condicted in the United States. All participants began receiving olanzapine treatment when they entered the study. The patients were then randomly assigned to an intervention group or a standard care group. Over the next 4-months, the intervention group participated in weekly psychoeducation classes focused on nutrition, exercise, and living a healthy lifestyle. Patients were followed for an additional 2 months to assess weight change. FINDINGS: A statistically significant difference in weight change between the two groups was observed post-treatment and at endpoint. At endpoint, the mean weight change of the intervention group was -.06 pounds, while the mean weight change in the standard care group was 9.57 pounds. In both groups, men gained significantly more weight than did women. CONCLUSIONS: The results indicate that a structured educational intervention might have a positive effect on antipsychotic-induced weight gain among patients with schizophrenia. PMID- 14562492 TI - Measuring continuity and discontinuity following stroke. AB - PURPOSE: To design and test a valid, reliable, and parsimonious instrument to measure continuity and discontinuity of self following stroke. DESIGN: A 45-item scale with a 3-point response set was developed. The conceptual basis, "continuity and discontinuity in the experience of self," was derived from an existential-phenomenological study of stroke survivors. Study instruments included the Barthel Index (measure of functional abilities) and the pre- and poststroke QoL Cantril Ladder. METHODS: The instruments were administered one time, in random order, to 55 stroke survivors in the United States in rehabilitation hospitals and in the community. Data were analyzed using a principal components factor analysis. RESULTS: A 2-factor solution with 10 items per factor was derived. Cronbach's alpha for Factor 1 (discontinuity of self) was .874 and for Factor 2 (continuity of self) was .869. Continuity and discontinuity were inversely related; continuity was related to functional abilities, but discontinuity was not. CONCLUSIONS: Use of this instrument validated findings from a qualitative study, which showed that life following stroke was a paradoxical sense of both continuity and discontinuity in the experience of self. This instrument can be used to measure the effectiveness of nursing interventions aimed at increasing stroke survivors' sense of continuity. PMID- 14562493 TI - Nurses' attitudes and practice related to hospice care. AB - PURPOSE: To describe characteristics, attitudes, and communications of nurses regarding hospice and caring for terminally ill patients. DESIGN: A cross sectional study of randomly selected nurses (n = 180) from six randomly selected Connecticut community hospitals was conducted in 1998 and 1999. METHODS: Hospice related training, knowledge and attitudes, demographic and practice characteristics, and personal experience with hospice were assessed with a self administered questionnaire (response rate = 82%). Logistic regression was used to model the effects of hospice-related training, knowledge, and attitudes on these outcomes, adjusting for personal experience and other characteristics of nurses. FINDINGS: Characteristics associated with discussion of hospice with both patients and families included greater religiousness, having a close family member or friend who had used hospice, and reporting satisfaction with hospice caregivers. Greater self-rated knowledge was significantly associated with discussion of hospice with patients. Attitudinal scores indicating greater comfort with initiating discussion and greater perceived added benefit of hospice were significantly associated with discussion with patients' families. CONCLUSIONS: Nurses' discussion of hospice with terminally ill patients and their families are related to the potentially modifiable factors of self-rated knowledge and attitudes revealing comfort with discussion and perceived benefit of hospice care. PMID- 14562494 TI - Views of community-dwelling, old-old people on barriers and aids to nutritional health. AB - PURPOSE: To examine barriers and aids to maintaining the nutritional health of community-dwelling old-old residents from their perspectives. DESIGN: Cross sectional exploratory study completed in March 2002 of old-old people living independently in one county in the Midwestern United States (US). METHODS: Sixty eight in-home interviews were conducted with community-dwelling people aged 80 or older. Content analysis was done to examine answers to two open-ended questions about barriers and helps to nutritional health. FINDINGS: These old-old people believed they were doing well nutritionally despite reduced independence and physical limitations. They were positive about their lives and creative in problem solving to remain independent. Social connectedness was the major factor for maintaining independence into old age. The leading barrier to maintaining nutritional health was health problems. Those with more barriers were more likely to be depressed. CONCLUSIONS: Knowledge of aids and barriers to nutritional health, from a personal perspective, gives an understanding of the issues and concerns of old-old people. PMID- 14562496 TI - Barriers to utilization of prenatal care services in Turkey. AB - PURPOSE: To identify barriers to utilization of prenatal care services in Turkey, including pregnant women's attitudes toward pregnancy and prenatal care. DESIGN: Descriptive. The population was Turkish women who lived in Erzurum and had delivered their infants but were still hospitalised. METHODS: The sample of 446 women had or had not received prenatal care, had no complications during pregnancy, carried their pregnancies to term, and were considered to have normal deliveries. Attitudes toward pregnancy and prenatal care and barriers to prenatal care services were measured by use of a questionnaire. FINDINGS: Low education of pregnant women and unwanted pregnancy were barriers to use of prenatal care services. Additional barriers were negative attitudes toward pregnancy and attitudes toward prenatal care. These barriers decreased frequency of use and delayed early initiation of prenatal care. The most important barrier reported by the women was being too busy at home to seek care. CONCLUSIONS: Although this sample was limited, the findings indicate barriers for attention by health care providers to ensure appropriate prenatal care and maternal and infant health. PMID- 14562495 TI - A theoretical explanation for previously infertile mothers' vulnerability to depression. AB - PURPOSE: To present a theoretical approach to understanding the potential vulnerability to depression in previously infertile new mothers. METHODS: This approach was developed by synthesizing results of a series of grounded theory studies of experiences of infertility and pregnancy and of parenting after infertility. These results were further synthesized with the relational cultural theory (RCT). FINDINGS: Previously infertile new mothers have experienced repeated and sustained interferences with significant relationships over the course of their infertility, which could lead to depression. CONCLUSIONS: Implications of this theoretical perspective include: (a) it is a new approach to understanding a significant and sometimes minimized issue in women's health; (b) it is a basis for designing an intervention for women coping with infertility and its aftermath; and (c) it might be applied to other women's health issues. PMID- 14562497 TI - The nurse-community health advocate team for urban immigrant primary health care. AB - PURPOSE: To describe: (a) development and implementation of an urban outreach health program for Latino immigrants; (b) nurse-community-health advocate (CHA) partnership roles in primary health care delivery, and (c) lessons learned from these activities over 7 years in urban community settings. METHODS: Descriptive study of a community-based health project in a large Midwestern American city. Information was gathered from participants and staff, from observing staff, and from a variety of sources to describe the program and its individual, family, and community effects. FINDINGS: Major findings pertain to the project team's ability to address the health promotion needs of Latino immigrant families and to successfully incorporate CHAs in planning and implementing the program. CHAs were a "bridge" between health programs and the community, promoting cultural sensitivity. CHAs and nurses provided a range of services including health education and promotion, outreach through home visits, assessment of family needs for referrals to appropriate resources, and follow-up support. CONCLUSIONS: The nurse-CHA team was an effective strategy for promoting Latino immigrant families' access to needed health care. This framework allowed for flexibility in assisting clients of different cultural backgrounds to obtain appropriate health care. PMID- 14562498 TI - Parental consent and adolescent risk behavior research. AB - PURPOSE: To identify methodological issues related to the use of active or passive parental consent in school-based research on adolescent risk behavior research and to propose recommendations consistent with current legal and ethical standards in the United States. METHODS: Review and synthesis of the professional literature related to adolescents and parental consent, federal regulations and guidelines in the United States, and the author's experience presenting these arguments and issues to institutional review boards and funding agencies for over 10 years. FINDINGS: The procedures used for parental consent affect a study's participation rates, costs, and selection bias. When active parental consent is required, parental permission is typically obtained for only 30%-60% of students, compared to 93%-100% when passive consent is used. Extensive follow-up may result in 55%-100% of parents giving permission, but at significant cost (typically $20 $25 per student). Active consent results in the exclusion of minorities, students having problems in school, and students already engaged in or at risk for problem behaviors. Strong methodological reasons were identified for using passive parental consent procedures when possible. Current federal regulations include four areas for possible waiver or alterations in parental consent procedures, including the use of passive parental consent. CONCLUSIONS: Health researchers must understand the methodological, legal, and ethical issues related to parental consent to produce high-quality, valid research about adolescents and to provide evidence for laws, policies, and regulations. PMID- 14562499 TI - Professional values held by baccalaureate and associate degree nursing students. AB - PURPOSE: To determine the congruency in value orientation of graduating students in baccalaureate and associate degree programs. DESIGN AND METHODS: A survey design with a convenience sample of 1,450 graduating nursing students from all baccalaureate and associate degree nursing programs in Texas. Data were collected using the Nurses Professional Values Scale (NPVS). Descriptive and parametric statistics were used for analysis. FINDINGS: ADN and BSN students did not differ significantly on the NPVS total score, however, ADN student scored higher on 5 of the 11 subscales than did their BSN counterparts. Men from both programs scored significantly lower than did women on the total scale and all subscales. Ethnic groups differed on the responses to three of the subscales representing nurses' values: respect for human dignity, safeguarding the client and public, and collaborating to meet public health needs. CONCLUSIONS: Professional values in graduating nursing students were significantly related to sex and ethnicity, regardless of educational program. Nursing faculty members are challenged to address these differences during the educational process and mentoring of students. PMID- 14562500 TI - [General statement about birth, consensus proposal concerning pain during childbirth]. PMID- 14562501 TI - [Difficulties in breast feeding]. PMID- 14562502 TI - [Support breast feeding in welfare (PMI--Protection Maternelle et Infantile)]. PMID- 14562503 TI - [Lactation consulting]. PMID- 14562504 TI - [Breast feeding and culture]. PMID- 14562505 TI - [Breast feeding and working]. PMID- 14562506 TI - [Obstacles and benefits of breast feeding the premature baby]. PMID- 14562507 TI - [How to use the breast pump]. PMID- 14562508 TI - [Found her Everest in Lapland]. PMID- 14562509 TI - [4/6 oxygen therapy]. PMID- 14562510 TI - [II-- Proper role of nurseries in infant care]. PMID- 14562511 TI - Don't send it yet! Getting your manuscript ready to submit. AB - Following these steps to finalize your paper will make a big difference in the overall reaction of the reviewers and editors. You are not only more likely to get an acceptance, but you also are more likely to be pleased when you read the article again, when it is in print! PMID- 14562512 TI - Writing a feature article: not all articles are alike. PMID- 14562513 TI - Manuscript submitted elsewhere. PMID- 14562514 TI - Sensing and signaling DNA damage: roles of Rad17 and Rad9 complexes in the cellular response to DNA damage. PMID- 14562515 TI - Stem cells in the mammalian blastocyst. PMID- 14562516 TI - How the ear's works work: mechanoelectrical transduction and amplification by hair cells of the internal ear. PMID- 14562517 TI - The p53 saga: the good, the bad, and the dead. PMID- 14562518 TI - Genomics approaches to photoreceptor development and disease. PMID- 14562519 TI - The group therapist's shame: a much undiscussed topic. AB - The topic of shame in group therapy has received limited attention in the group therapy literature. When the topic has been addressed, the focus has been on the shame of the group members. The shame of the group leader and its effect on leadership efficacy and group process has received inadequate attention, given what seems to be its power and prevalence. In this article we examine shame and the group therapist with regard to (1) potentially shameful topics and (2) dynamics that evoke shame. Suggestions for both the mitigation of shame and the enhancement of the leader's self-esteem in dispatching the role and functions of group therapist are offered. In discussing this subject, we propose to normalize group therapist shame by emphasizing the challenges and complexity of group leadership. While the examples illustrate an array of emotions, our purpose is to focus on their shameful elements because they are (1) frequently ignored or overlooked; (2) not made explicit; or (3) discussed in other terms, such as narcissism. In doing so, we invite the reader to consider the clinical utility of the hypothesis that shame has a powerful presence and impact on our leadership image and effectiveness. PMID- 14562520 TI - NEO-five factor personality traits as predictors of response to two forms of group psychotherapy. AB - The relationships between patient personality variables and outcome for 107 psychiatric outpatients with complicated grief who completed either interpretive or supportive short-term group therapy were investigated. The personality variables were assessed prior to treatment with the NEO-Five Factor Inventory (NEO-FFI). For patients in both forms of therapy, extraversion, conscientiousness, and openness were directly associated with favorable treatment outcome. In contrast, neuroticism was inversely related to favorable outcome for patients in both forms of therapy. Agreeableness was directly related to favorable improvement in grief symptomatology for patients in interpretive therapy, but not for those in supportive therapy. The results highlight the importance of assessing patient personality in order to predict response to short term group therapy. Possible explanations and clinical implications of these findings are discussed. PMID- 14562521 TI - Eroticism in group psychotherapy: psychoanalytic reflections on desire, agony, and ecstasy. AB - To fully understand the complexities of eroticism in groups, it may be necessary to review a conceptual differentiation of desire and its allies: agony and ecstasy. This article suggests that psychoanalytic group psychotherapy is made for neither agony nor ecstasy. Sexual excitement maybe; eroticism and desire, yes; agony and ecstasy, no. While agony or ecstasy imply a threat to the survival of the group, eroticism and desire reaffirm its existence. In this manner the group may be converted into a theater where desire may be celebrated, while the threat of being dissolved in the depths or exaltation of agony and ecstasy is elaborated and worked through: "Desire is desire only if it succeeds in postponing something". PMID- 14562522 TI - Pursuing relational consciousness: thinking and antithinking in group. AB - Although human beings are interpersonal and innately curious, at the same time an aspect of the self defends against mental relationships with self and other, because such relationships threaten to cause psychic pain. Bion's ideas on thinking and antithinking are applied to this topic of relational consciousness: A "psychotic part of the personality"--which refers also to the "basic assumptions" level of the group-prefers antithinking: forestalling, evading, and assaulting thinking and thinkers. Clinical examples illustrate how the group therapist may address and treat the pervasive resistances to relational consciousness in the group setting. These involve understanding the dynamics and clinical manifestations of hatred of thinking, excessive projective identification, anticipatory anxiety regarding thinking, and bizarre and hallucinatory thought transformations. PMID- 14562523 TI - Working through a psychotherapy group's political cultures. AB - Macropolitical evolution, starting with authoritarian monarchism, has moved through anarchistic transitions either to the totalitarianism of fascism and communism or to liberal and social democracy. We posit analogous micropolitical development in process-oriented therapy groups: "dependence" and "counterdependence" corresponding to monarchism and anarchism; and "independence" and "interdependence" to liberal and social democracy, respectively. Transition from counterdependence to independence and interdependence may be: (1) facilitated through group members' cooperative experience of rebellion, or (2) blocked by collective identification, the internalization of dystopian or utopian fantasies that coalesce as "group-self" perceptions. We explore how group therapists work clinically with and through these several "political cultures" in the service of group and self transformation. PMID- 14562524 TI - Asymptomatic bacteriuria among outpatients with diabetes mellitus in an urban black population. AB - OBJECTIVE: To determine the prevalence of asymptomatic bacteriuria (ASB) in individuals afflicted by Diabetes mellitus; the antibiotic susceptibility of the microbial isolates and the association of host factors with ASB. DESIGN: This was a prospective cross sectional study. SETTING: Attendants of outpatient polyclinics at three main tertiary hospitals; namely, Harare, Chitungwiza and Parirenyatwa Hospitals. SUBJECTS: 176 participants. MAIN OUTCOME MEASURES: Patients attending the polyclinics between 6.30 am and 9.30 am from Monday to Friday were randomly selected. Demographic data was obtained at enrollment using a standardized questionnaire. Fasting venous blood was withdrawn from the participants for glucose analysis. Clean-catch midstream urine samples from all men and women were cultured and the causal organisms were isolated and identified by standard microbiological methods. Antibiotic susceptibility testing was performed using a disc diffusion method. Potential host factors included age, type of diabetes, duration of diabetes, glucosuria and leukocyturia. RESULTS: The prevalence of ASB was 32% in the diabetics and 11% in nondiabetic participants. The commonest bacterial organism isolated in participants afflicted by Diabetes mellitus was Escherichia coli (26%) followed by Staphylococcus aureus (21%), Streptococcus group B (14%), Streptococcus group D and non-lactose fermenting coliforms (7% respectively). Other isolates were Micrococcus and Pseudomonas (5% respectively), Klebsiella and Proteus (2% respectively). Gentamicin, nitrofurantoin, ampicillin and nicene were the most effective antimicrobials in the majority of isolates. Certain isolates exhibited some bacterial resistance to conventional antibiotics. Of the host factors, an association was found between bacteriuria and glucosuria (p < 0.001) and between leukocyturia and bacteriuria (p = 0.005). CONCLUSION: The prevalence of ASB is increased in diabetes and the rather high blood glucose levels exhibited by these individuals may further complicate this condition. As some bacterial species exhibited resistance to some common antimicrobials, these results raise questions regarding future clinical reliability of some conventional antimicrobials when considering therapy for asymptomatic bacteriuria. PMID- 14562525 TI - Lipoprotein(a) concentrations and apolipoprotein(a) isoform distribution in a Zimbabwean population. AB - OBJECTIVE: To determine serum lipoprotein(a)[Lp(a)] concentrations and apolipoprotein(a)[apo(a)] phenotypes in a Zimbabwean population. DESIGN: Cross sectional study. SETTING: Blood Transfusion Services, Harare, Zimbabwe. SUBJECTS: 84 black and 40 white blood donors. MAIN OUTCOME MEASURES: Lp(a) concentrations and apo(a) phenotypes. RESULTS: The mean and median values for Lp(a) concentrations were 506 and 350 mg/L for the black subjects and 278 and 142 mg/L for the white subjects (p < 0.005). The frequency distributions of Lp(a) concentrations for both populations were skewed to the right. The frequency distribution of apo(a) size, expressed as the number of kringle IV repeats, was determined. Comparison of the frequency distribution plots showed very similar isoform distributions between the two groups. The documented inverse relationship between apo(a) size and Lp(a) concentration was observed in the white population. CONCLUSION: The Lp(a) levels in the black population were two to three fold higher than in the white population whilst no differences in apo(a) phenotype distribution were noted. This suggests that environmental and metabolic factors may be responsible for the elevated Lp(a) levels observed in blacks. Thus different pathological thresholds may have to be established for elevated serum Lp(a) levels to be used as a risk marker for coronary heart disease in black populations. PMID- 14562526 TI - The acceptability of insecticide treated mosquito nets among community members in Zimbabwe. AB - OBJECTIVES: To assess the acceptability of insecticide treated mosquito nets (ITNs) among community members in Zimbabwe for the implementation of effective malaria intervention using ITNs. DESIGN: A cross sectional study and longitudinal study were carried out using questionnaires, which were administered by health workers from rural health centres. SETTING: Communities in rural areas from seven districts namely Bulilimamangwe, Chipinge, Gokwe, Hurungwe, Lupane, Mount Darwin and Uzumba Maramba Pfungwe (UMP) in Zimbabwe. SUBJECTS: 1,576 community members who bought ITNs in the project areas. MAIN OUTCOME MEASURE: Acceptance of ITNs and price, reasons for wanting or not wanting nets; ITNs usage and washing pattern among respondents. RESULTS: More than 90% of people said they bought the nets from the project in order to avoid mosquito bites and to prevent malaria. Quite a number of respondents (69%) gave the answer that they bought mosquito nets from the project because they were cheaper (Z$130 to Z$390) than the market price (Z$1,000) and more than half (58.3%) claimed that they could not afford to buy mosquito nets at the market price. The price was accepted by the majority as cheap or very cheap in Gokwe, Hurungwe, Mount Darwin, and UMP (92%, 82%, 98% and 90%, respectively). Other responses given for having bought nets wereat health staff's recommendation (71.3%), and they were suffering from malaria (40.9%). The reasons for not having bought mosquito nets were that they had not suffered from malaria (55.5%), that they did not like to use a mosquito net (31.1%), did not know the efficacy of the net (21.6%), or there were not many mosquitoes at home (28.9%). The percentage of ITN use was very high (90 to 100%) amongst those who bought nets. The percentage of children under five years and pregnant women who were ITN users ranged from six to 24% to 2.8 to 9.7%, respectively. Percentage of "others" which means those above five years and not pregnant was more than 70% in all the districts. The washing pattern of ITNs in Chipinge showed that most of the mosquito nets (74.5%) were washed every three to five months. CONCLUSION: Acceptability of ITNs was very high based on reported utilisation of ITNs. It is recommended that rural communities should be sensitised on the importance of treated nets for malaria prevention, and advised to purchase mosquito nets whenever cash is readily available in each community. PMID- 14562527 TI - Nephroblastoma in horseshoe kidney. AB - A case of nephroblastoma arising in a horseshoe kidney is reported. Clinically a bilateral nephroblastoma was suspected. Pre-operative diagnosis of horseshoe kidney, once a mass has arisen, may be difficult even with radiological investigation. PMID- 14562528 TI - Clinical and biomedical aspects of gonorrhoea, diagnosed in symptomatic patients in Midlands Province, Zimbabwe. PMID- 14562529 TI - Effects of a herbal remedy (CATAR) on human cataract lenses: observations from a preliminary experimental study. PMID- 14562530 TI - Comparative efficacy of chloroquine and cotrimoxazole in the treatment of acute uncomplicated falciparum malaria in Nigerian children. AB - OBJECTIVES: To evaluate the efficacy of two dosing regimens of cotrimoxazole in the treatment of falciparum malaria and compare the efficacy with that of chloroquine, the first-line antimalaria drug in the area of study. DESIGN: A prospective cross sectional study. SETTINGS: Department of Clinical Pharmacology, University College Hospital, Ibadan, Nigeria. SUBJECTS: Ninety eight children with acute symptomatic uncomplicated falciparum malaria. MAIN OUTCOME MEASURES: Fever and other symptoms clearance times, parasite clearance times and the cure rates for chloroquine, three day and five day cotrimoxazole. RESULTS: Ninety eight children with acute symptomatic uncomplicated falciparum malaria were randomised to receive three doses of oral chloroquine and two regimens of cotrimoxazole. Pre-treatment clinical and parasitological parameters were similar in the three treatment groups. The fever and other symptoms clearance times were comparable in all the treatment groups: 1.83 +/- 1.3, 1.9 +/- 1.0 and 2.4 +/- 1.3 days for chloroquine, three day cotrimoxazole and five day cotrimoxazole, respectively p = 0.24. Parasite clearance times for the three treatment groups were also similar; 3.0 +/- 1.0, 3.1 +/- 0.7, and 3.0 +/- 1.0 days respectively for chloroquine, three day- and five day- cotrimoxazole; p = 0.96. The cure rates for chloroquine, three day and five day cotrimoxazole were 74.2%, 88.2% and 84.8%, respectively (x2 = 2.40, p = 0.30). The three treatment regimens were well tolerated. CONCLUSION: These results indicate that cotrimoxazole is as effective as chloroquine in treatment of acute symptomatic uncomplicated falciparum malaria in children resident in an endemic area of southwest Nigeria. It is an added advantage when malaria coexists with respiratory tract infections for which cotrimoxazole is the recommended drug. PMID- 14562531 TI - A comparison of topical Phenytoin with Silverex in the treatment of superficial dermal burn wounds. AB - OBJECTIVE: To compare topical diphenylhydantion (Phenytoin) with silver sulphadiazine/chlorhexidine (Silverex) in terms of rate of wound healing, analgesic and antibacterial properties in small to moderate-sized (< 30% TBSA) superficial dermal (second degree) burn wounds. DESIGN: A prospective randomized controlled study. SETTING: Surgical wards, Muhimbili National Hospital from July 2000 to February 2001. SUBJECTS: Sixty four patients with acute burns, 32 in each group. INTERVENTIONS: Study group treated by sprinkling Phenytoin powder and control group by sprinkling Silverex powder on the wounds for 14 days or until the wound epithelialised or was ready for skin grafting. The data collected included demographic characteristics of patients, aetiology of burn injury, circumstances of injury, site and extent of burns, pus discharge and smell from the wound, pain and discomfort from the wound, bacterial cultures of wound swabs, rate of reduction in wound size and outcome of treatment. RESULTS: The study enrolled 33 male and 31 female patients, 69% being children under five years of age. Hot liquids (80%) and open flames (20%) were the only causes of burns. In 97% of patients injury was due to domestic accidents. In half of the patients burns involved the trunk, and 52% of all patients had less than 15% total body surface burnt. Pus discharge was recorded in 59% of Phenytoin-treated and 75% in Silverex-treated patients while foul smell was noted in 19% and 31% of cases respectively. There were more negative bacterial wound cultures in Phenytoin-than Silverex-treated wounds on day five and day 10 of treatment, the difference being statistically significant (p < 0.01 and 0.001 respectively). There was also a statistically significant difference in wound pain in favour of Phenytoin (p < 0.01). There was no statistically significant difference in the rate of healing in the two groups. CONCLUSION: Phenytoin is a cheap and easy-to-use medicament, effective in suppressing burn wound bacteria and relieving pain thereby promoting healing, and may be advocated for the purpose in resource-scarce environments. PMID- 14562532 TI - Jaw fractures in Nigerian children: an analysis of 102 cases. AB - OBJECTIVES: To present the prevalence and pattern of jaw fractures in children aged 15 years and below attending two hospitals serving as referral centres for facial injuries in North Eastern Nigeria. DESIGN: Retrospective Cross Sectional Study. SETTINGS: Department of Oral and Maxillofacial Surgery, University of Maiduguri Teaching Hospital, Borno State, Nigeria. SUBJECTS: One hundred and two patients with jaw fractures aged 15 years and below. MAIN OUTCOME MEASURES: Aetiological factors and pattern of jaw fractufes in children aged 15 years and below. RESULTS: We retrospectively reviewed 102 patients with jaw fractures aged 15 years and below seen over a five year period at two referral centres in Maiduguri, Northeastern Nigeria. This constituted 9.5% of the total 1,074 cases of maxillofacial injuries managed during the period. The male to female ratio was 7.5:1 and there was a male responderance in all age groups. The main causes of fractures were road traffic accident (n = 55, 53.7%) followed by falls (n = 26, 25%). Other etiological factors such as fights, sports and gunshots accounted for the remaining cases. The mandible was the commonest site with the body commonly fractured (n = 74, 72.5%). No Le Fort III fractures were identified in this study. CONCLUSION: Despite some limitations to our data, such as exclusion of patients who attended private clinics and lack of modern diagnostic methods, this report shows that there is obvious need for concern about the high prevalence of maxillofacial injuries in children caused by road traffic accidents in North Eastern Nigeria. PMID- 14562533 TI - Awareness of the risks of HIV infection by Zimbabwean urban and rural high school attendees. AB - OBJECTIVES: The purpose of this report was to investigate the awareness by high school attendees of risky behaviour likely to lead to sexually transmitted infections (STIs) including HIV/AIDS and to assess their preparedness to prevent or obviate the consequences. DESIGN: Cross sectional study. SETTINGS: A small town located 100 km south of the Zimbabwean capital, Harare and in communal and commercial farming areas within a 50 km radius of the town. METHODS: With the permission of school authorities, a group of second year medical students from the University of Zimbabwe on rural attachment administered an anonymous questionnaire to teenagers attending four high schools in the study area. Specific questions to determine whether the teenagers were sexually active and if they used any protection during sex were incorporated into a general and environmental health questionnaire. RESULTS: A total of 241 teenagers, 153 boys and 89 girls from all four schools completed the questionnaire. The percentages of sexually active boys (48%) and girls (49%) were similar. The majority had heterosexual preferences, 4.6% boys were homosexual. Half (50%) of the 16 year old pupils were sexually active. There were more urban than rural school attendees who reported personal knowledge of someone with HIV or AIDS. Condom use was low. CONCLUSION: We found that half of the 16 to 19 year old students were sexually active. Rural school attendees were more likely to be sexually active and less likely to practice safe sex. We conclude that the risk of sexually transmitted diseases (including HIV) amongst these teenagers is significant and recommend that, there is a need to design a group targeted awareness programme in order to obviate sexually transmitted diseases including HIV. PMID- 14562534 TI - The serum concentrations of zinc, copper and selenium in children with inflammatory bowel disease. AB - OBJECTIVE: To estimate the levels of trace elements in children with inflammatory bowel disease (IBD). DESIGN: Prospective cross sectional study. SETTING: Gastroentrology Unit, Great Ormond Street Children's Hospital, London, UK. SUBJECTS: Seventy four children with inflammatory bowel disease confirmed endoscopically and histologically (38 ulcerative colitis and 36 Crohn's disease) and 40 age matched controls had their serum zinc, copper and selenium assayed at presentation. MAIN OUTCOME MEASURE: Serum levels of zinc, copper and selenium in children with inflammatory bowel disease and age matched controls. RESULTS: Seventy four children with inflammatory bowel disease confirmed endoscopically and histologically (38 ulcerative colitis and 36 Crohn's disease) and 40 age matched controls had their serum zinc, copper and selenium assayed at presentation. The serum levels of selenium were significantly lower in cases of ulcerative colitis 0.63 +/- 0.25 mmol/L and Crohn's disease 0.69 +/- 0.25 mmol/L than in the controls 0.84 +/- 0.13 mmol/L (p < 0.01). The serum copper concentration was significantly higher in those with Crohn's disease 22.7 +/- 5.49 mmol/L than in those with ulcerative colitis 17.6 +/- 5.15 mmol/L and the controls 20.76 +/- 4.06 mmol/L (p < 0.01). Children with Crohn's disease had a lower serum zinc level 11.01 +/- 2.49 mmol/L compared to the control level of 13.6 +/- 1.63 mmol/L (p < 0.05), but the levels were not significantly different in the controls and ulcerative colitis (p > 0.10). Children with inflammatory bowel disease have abnormal levels of the trace elements which is more marked in those with Crohn's disease. CONCLUSION: Children with IBD in this study show abnormalities of the trace elements which is probably a result of inadequate intake, reduced absorption, increased intestinal loss due to impairment of the absorption as a result of the inflammatory process. The reduced free radical scavenging action of zinc and selenium as a result of their deficiency may contribute to the continued inflammatory process of IBD. The recommendation of the supplementation of these trace elements in IBD is further supported by the findings of this study in children. PMID- 14562535 TI - [Interventional MRT]. PMID- 14562536 TI - [Clinical use of interventional MR imaging]. AB - The integration of diagnostic and therapeutic procedures by MRI is based on the combination of excellent morphologic and functional imaging. The spectrum of MR guided interventions includes biopsies, thermal ablation procedures, vascular applications, and intraoperative MRI. In all these applications, different scientific groups have obtained convincing results in basic developments as well as in clinical use. Interventional MRI (iMRI) is expected to attain an important role in interventional radiology, minimal invasive therapy, and monitoring of surgical procedures. PMID- 14562537 TI - [Safety and safety limits in MRI]. AB - The influence of electromagnetic fields on humans through the use of magnetic resonance tomography is an ongoing topic in the scientific literature. However, the contents of relevant standards and legal regulations governing the operation of MR equipment are less well known. We summarize the actual status and give an overview of the national and international bodies in charge of generation and publication of guidelines and regulations. PMID- 14562538 TI - [Artifacts in MRT caused by instruments and implants]. AB - Metallic instruments and implants can cause severe image artifacts in magnetic resonance imaging (MRI). Besides the properties of the materials and the geometrical arrangement of the devices, the applied MRI sequence type and its parameters (echo time, voxel size, read-out bandwidth, orientations of encoding directions, etc.) play also an important role. These interactions are presented in a systematic survey. A detailed description of the basic physical mechanisms underlying the generation of artifacts is also provided. PMID- 14562539 TI - [An approach for safe visualization and localization of catheter during MR-guided intravascular procedures]. AB - The present paper describes a method for the visualization and localization of a catheter during real-time imaging on a clinical magnetic resonance tomograph (MRT). This method includes the use of an apposite catheter hardware equipped with an optically tunable resonant marker on its tip (OptiMa-Catheter). In the tuned state, the marker produces an intense signal spot in the MR image. The signal is modulated by optical detuning controlled by the MRT system, which results in improved visualization of the catheter tip due to flashing. During real-time imaging, additionally, the localization of the tip is precisely detected and the slice tracking automatically performed. Experiments demonstrate that the OptiMa-Catheter does not generate relevant resonance heating during imaging. Consequently, it combines the visualization and localization known from active techniques with the safety of passive techniques. PMID- 14562540 TI - [Automatic slice tracking in interventional magnetic resonance imaging]. AB - Magnetic resonance imaging (MRI) is ideally suited to monitor minimally invasive operations with catheters or needles, since it offers both a superior soft-tissue contrast and the possibility to perform functional tests. In the present study, small radio-frequency coils were attached to the instruments in order to localize the MR-invisible instruments. The implementation of active instrument tracking is described on the basis of the example of active catheter tracking. In this case, the current position information of the instrument is used to automatically position the MRI slice at the catheter location. In combination with a user interface, the interventional radiologist is offered the possibility to perform vascular interventions from within the MR scanner room. At image update rates of approximately 3 Hz, tracking and placement of catheters in vascular structures are possible with interactive switching of slice orientation and image contrast. In an animal model, the technique was successfully used to selectively visualize the abdominal vessels and their branches under MRI guidance. PMID- 14562541 TI - [Comparison of noninvasive MRT procedures for temperature measuremnt for the application of medical heat therapies]. AB - Novel methods for hyperthermia tumor therapy, such as high-intensity focused ultrasound (HIFU) or laser-induced thermotherapy (LITT), require accurate non invasive temperature monitoring. Non-invasive temperature measurement using magnetic resonance imaging (MRI) is based on the analysis of changes in longitudinal relaxation time (T1), diffusion coefficient (D), or water proton resonance frequency (PRF). The purpose of this study was the development and comparative analysis of the three different approaches of MRI temperature monitoring (T1, D, and PRF). Measurements in phantoms (e.g., ultrasound gel) resulted in the following percent changes: T1-relaxation time: 1.98%/degree C; diffusion coefficient: 2.22%/degree C; and PRF: -0.0101 ppm/degree C. All measurements were in good agreement with the literature. Temperature resolutions could also be measured from the inverse correlation of the data over the whole calibration range: T1: 2.1 +/- 0.6 degrees C; D: 0.93 +/- 0.2 degree C; and PRF: 1.4 +/- 0.3 degrees C. The diffusion and PRF methods were not applicable in fatty tissue. The use of the diffusion method was restricted due to prolonged echo time and anisotropic diffusion in tissue. Initial tests with rabbit muscle tissue in vivo indicated that MR thermometry via T1 and PRF procedures is feasible to monitor the local heating process induced by HIFU. The ultrasound applicators in the MR scanner did not substantially interfere with image quality. PMID- 14562542 TI - [Interventional MRA: concepts for active visualization of catheters and stents]. AB - A precondition to safe guidance of vascular guidewires and catheters during the course of magnetic resonance (MR)-guided vascular intervention is a high-contrast visualization of the instruments. The integration of miniature radiofrequency (RF) coils and coaxial cables into guidewires and catheters enables the reception of RF signal from the lumen of blood vessels, and thus the active visualization of the instruments. Moreover, metallic vascular implants (stents) can be modified to act as intravascular RF antennas that inductively couple their RF signal to a conventional surface RF coil. Such stent resonators show signal amplification inside the lumen of the stent and thus can be visualized with high contrast in MR images. Furthermore, once such a device has been implanted, the method offers the potential for non-invasive long-term follow-up of the stent patency. PMID- 14562543 TI - [MRI-guided surgery with high intensity focused ultrasound]. AB - High-intensity focused ultrasound allows high-precision, non-invasive thermocoagulation of tissues within seconds, with sparing of surrounding areas. The resulting tissue necrosis is so sharply demarcated that the technique is also defined focused ultrasound surgery (FUS). The combination with magnetic resonance imaging (MRI) allows an exact definition of the target volume and a safe guidance of FUS. The present paper describes the physical equipment necessary to perform MRI-guided FUS, and reports an example of application of this technique for the therapy of breast cancer. Finally, the paper outlines further examples of FUS application and future perspectives. PMID- 14562544 TI - [Interventional MR-mammography: manipulator-assisted large core biopsy and interstitial laser therapy of tumors of the female breast]]. AB - The present study investigated the clinical application of magnetic resonance (MR)-guided breast interventions, such as manipulator-assisted large core breast biopsy (LCBB) inside a 1.5 T whole-body magnet and MR-guided interstitial laser therapy (ILT). Sixteen patients underwent LCBB and 1 additional patient underwent 4 sessions of ILT of a recurrent undifferentiated lymph node metastasis in the axilla using a Nd-YAG laser (1064 nm). Temperature changes of the tumor tissue during ILT were monitored using phase images of a gradient echo sequence (GRE) (TR/TE/FA = 25/12/30). In 5 patients the biopsy findings were histopathologically confirmed after open surgery. In 3 patients, the biopsy missed one tubular and one ductal carcinoma; one invasive carcinoma was underestimated. Eight patients with benign findings are still in the follow-up period. The heating zone during ILT was well delineated on subtracted phase images. No severe adverse events were observed with LCBB or ILT. MR-guided breast biopsies are feasible with the manipulator system inside a whole-body 1.5 Tesla MR scanner. GRE information is suitable for therapy monitoring during ILT within the tumor. Further studies are necessary to evaluate the accuracy of the manipulator system and the efficacy of ILT in the treatment of breast lesions. PMID- 14562545 TI - [Cryotherapy of the brain--a new methodologic approach]. AB - Cryodestruction of tissue is influenced by cooling and thawing rates, absolute tissue temperature, number of freeze-thaw cycles, and type of tissue. However, under clinical conditions a MRT visualization of the temperature distribution during cryo-procedures is not possible. Thus, the extent of necrotic areas within the cryo-influenced regions are not precisely predictable. This limitation is particularly relevant for the application of cryoablation in the brain. The present paper proposes the concept of a local, cryo-induced ischemic necrosis. The basic concept is that the MRT-observable and surgically well-manageable frozen region is ischemic. This cryo-induced ischemia causes a necrosis. The extent of the necrotic region is exclusively determined by the ischemia tolerance of the tissue. The effectiveness of this method is demonstrated on sheep brain in vivo. Compared to the freeze-thaw method, histological examinations show a sharper demarcation between regions of necrosis and healthy tissue. In conclusion, the method of MR-controlled local, cryo-induced ischemia enables an exact definition of the region of necrosis in the brain. PMID- 14562546 TI - [MR-guided laser-induced thermo-ablation of liver tumors: clinical experiences and therapy control concepts]. AB - Minimally-invasive, laser-induced interstitial thermotherapy (LITT) of solid tumors represents a valid alternative to surgical procedures such as tumor resections. Within the framework of a palliative study on 16 patients, a total of 25 metastases in the liver were treated in an open MR system (0.5 T). The intraoperative scanner design allows patient-based navigation, decisive for a safe applicator positioning, as well as temperature monitoring and direct inspection of the therapy result, without need for patient transfer or repositioning. Although the MR thermometry applied in the open scanner assisted LITT monitoring, the current accuracy of temperature data was not sufficient to serve automatic irradiation control. Therefore, an experimental monitoring and control system was developed in a closed MR scanner (1.5 T) featuring a calibrated MR thermometry. The system provides also an interface to the laser system, allowing the automatic off/on switching of the laser power according to preoperatively defined control criteria. The basic functionality of the automatic laser control was successfully demonstrated with laser ablation experiments of liver samples using irradiation parameters close to typical clinical values. PMID- 14562548 TI - [Finding the right keys]. PMID- 14562547 TI - [Intraoperative use of an open mid-field MR scanner in the surgical treatment of cerebral gliomas]. AB - The aim of the present study was to evaluate the effectiveness of intraoperative MRI guidance in achieving more gross-total resection in case of primary brain tumors. We studied 12 patients with low-grade glioma and 19 patients with high grade glioma who underwent surgery within a vertically open 0.5 T MR system. After initial imaging, the resection was stopped at the point in which the neurosurgeon considered the resection complete by viewing the operation field. At this time, intraoperative MRI was repeated ("first control") to identify any residual tumor. Areas of tumor-suspected tissue were localized and resected, with the exception of tissue adjacent to eloquent areas. Final imaging was carried out before closing the craniotomy. Comparison of "first control" and final imaging revealed a decrease of residual tumor volume from 32% to 4.3% in low-grade gliomas, and from 29% to 10% in high-grade gliomas. Intraoperative MRI allows a clear optimization of microsurgical resection of both low-grade and high-grade gliomas. PMID- 14562549 TI - [General conditions in 2003: psychiatry in crisis]. PMID- 14562551 TI - [From illusions to disappointment]. PMID- 14562552 TI - [The key of one meeting]. PMID- 14562550 TI - [The keys in psychiatry]. PMID- 14562553 TI - [The twist of the keys]. PMID- 14562554 TI - [The key to a meeting]. PMID- 14562555 TI - [Antipsychotic agents, the keys to the opening of treatment practices]. PMID- 14562557 TI - [The keys of knowledge, culture of change or doctrine of influence?]. PMID- 14562556 TI - [The keys to a quality relationship]. PMID- 14562558 TI - [Risk behavior in psychiatric emergency, crash by a trigger...]. PMID- 14562559 TI - [Methods in psychiatric care of victims of sexual aggression]. PMID- 14562560 TI - The systemic inflammatory response to cardiopulmonary bypass. AB - Although our understanding of the basic pathophysiology of systemic inflammatory response to CPB has significantly advanced in the last 2 decades, these experimentally derived ideas have yet to be fully integrated into clinical practice. Treatment of the systemic inflammatory response to CPB is also confounded by the fact that inhibition of inflammation might disrupt protective physiologic responses or result in immunosuppression. Although it is unlikely that no single therapeutic strategy will ever be sufficient in of itself to totally prevent CPB-associated morbidity, the combination of multiple pharmacologic and mechanical therapeutic strategies, each selectively targeted at different components of the inflammatory response, may eventually result in significantly improved clinical outcomes following cardiac surgery. PMID- 14562561 TI - Perioperative myocardial ischemia reperfusion injury. AB - Myocardial I-R injury contributes to adverse cardiovascular outcomes after cardiac surgery. The pathogenesis of I-R injury is complex and involves the activation, coordination, and amplification of several systemic and local proinflammatory pathways (Fig. 4). Treatment and prevention of perioperative morbidity associated with myocardial I-R will ultimately require a multifocal approach. Combining preoperative risk stratification (co-morbidity and surgical complexity), minimizing initiating factors predisposing to SIRS, limiting ischemia duration, and administering appropriate immunotherapy directed toward systemic and local proinflammatory mediators of I-R injury, should all be considered. In addition, the role of the genetic-environmental interactions in the pathogenesis of cardiovascular disease is also being examined. Thus, in the near future, preoperative screening for polymorphisms of certain inflammatory and coagulation genes should inevitably help reduce morbidity by permitting the identification of high-risk cardiac surgical patients and introducing the opportunity for gene therapy or pharmacogenetic intervention [42,64]. PMID- 14562562 TI - Neuroprotection in cardiac surgery. AB - This article reviews past and present neuroprotective efforts and outlines a framework for the future development of techniques for neuroprotection during cardiac surgery. PMID- 14562563 TI - Monitoring anticoagulation and hemostasis in cardiac surgery. AB - The need to monitor anticoagulation and hemostasis during and after cardiac surgery has led to recognition of the importance of evaluation and use of hemostasis monitors in this setting. Consequently, rapid and accurate identification of abnormal hemostasis has been the major impetus for the development of point-of-care tests and their use in transfusion algorithms for cardiac surgical and other critically ill patients. PMID- 14562564 TI - Current status of antifibrinolytics in cardiopulmonary bypass and elective deep hypothermic circulatory arrest. AB - Cardiopulmonary bypass (CPB) results in many physiologic derangements, including activation of the hemostatic and fibrinolytic pathways. Deep hypothermic circulatory arrest (DHCA) adds a further insult to the coagulation systems because it involves more extreme hypothermia and organ ischemia related to blood stasis. The abnormalities induced by CPB disrupt the checks and balances in the hemostatic and fibrinolytic systems, resulting in a pathologic state that leads to excessive bleeding and other perioperative complications. Prophylactic antifibrinolytic therapy can attenuate the response to this insult by restoring the delicate balance within these systems, potentially reducing the complication rate and improving patient outcomes. PMID- 14562565 TI - Clinical potential of blood substitutes or oxygen therapeutics during cardiac surgery. AB - Several complications and unforeseen adverse side effects have colluded to keep commercially available blood substitutes or oxygen therapeutic agents tantalizingly "just out of reach." Because the three classes of agents under development have different oxygen-delivery mechanisms and side-effect profiles, each can be expected to have its own unique clinical applications, particularly in the cardiac surgery population. The fact that South Africa recently approved one HBOC for use as a transfusion alternative in patients with chronic anemia indicates that initial clinical use is near for a number of these agents. It is to be hoped that they will be used for several applications rather than just for a "transfusion alternative." Despite some frustrating limitations, all of these agents are antigen and pathogen free, have an acceptable side-effect profile, and have a long shelf life. Increasing volunteer-blood-donor shortages, coupled with increasing blood-transfusion needs, and expanding human immunodeficiency virus and hepatitis B and C epidemics, continue to fuel the demand for further development of these products. Transfusion alternatives will eventually become commercially available--the question is "when," not "if." Equally important, these agents' potential for serving as effective oxygen-delivery agents to ischemic tissues heralds an entirely new field of clinical investigation. PMID- 14562566 TI - New treatments for perioperative cardiac arrhythmias. AB - Cardiac arrhythmias remain a major source of morbidity, mortality, and prolonged postoperative hospital stay in surgical patients. Recent studies in patients experiencing out-of-hospital cardiac arrest have expanded our knowledge in the management of cardiac arrhythmias. Future advances require additional studies focused on the unique proarrhythmic substrates in surgical patients, to provide a clear rationale for antiarrhythmic drug therapy in the perioperative period. PMID- 14562567 TI - What does transesophageal echocardiography add to valvular heart surgery? AB - No single monitoring tool in the last decade has had more of an effect on intraoperative decision making and surgical management of cardiac valvular pathologies than has TEE. It has become the standard of care for evaluating reparative valvular procedures, thus providing an immediate gauge of the surgical results and helping to avoid suboptimal surgical outcomes. As the technology of TEE and its application advance, so too should the ability to diagnose and manage valvular pathologies, broaden the range of surgical options, and ultimately improve patient outcomes. PMID- 14562568 TI - Off-pump coronary surgery: how do the anesthetic considerations differ? AB - In recent years, there has been much interest in performing coronary artery bypass graft (CABG) surgery without the aid of cardiopulmonary bypass (CPB). Initial efforts focused on "minimally invasive" direct coronary artery bypass, wherein the left anterior descending artery is bypassed with an in situ left internal mammary artery graft through a small left anterior thoracotomy. A more widely adopted approach however, is off-pump CABG (OPCAB), in which CABG surgery is performed on one or more vessels through the usual median sternotomy approach without the aid of CPB. This article reviews the differences in the anesthetic considerations of OPCAB compared to conventional CABG using CPB. PMID- 14562569 TI - Cardiac failure and left ventricular assist devices. AB - LVADs represent advanced therapy for cardiac failure. The anesthesiologist's contribution to the pre-, intra-, and postoperative management of these challenging patients continues to grow as LVAD technology is refined and as more patients become eligible to receive these life-saving devices. PMID- 14562570 TI - Anesthesia in the cardiac catheterization lab. AB - Because of the increasing need for anesthesia in the CCL, anesthesiologists are often involved in diagnostic and interventional procedures in this setting. Safe delivery of anesthesia requires adequate preparation and familiarity with the procedure and its surroundings. Radiation safety is of paramount importance for the patient and the practitioner, and ready access to the services of anesthesia support personnel, the pharmacy, and the stat laboratory is a key factor for success. In the CCL, and the traditional operating room environment, anesthesiologists shall continue to advance and improve patient care while reducing morbidity and mortality. PMID- 14562571 TI - Anesthetic management of the adult patient with congenital heart disease. AB - As the number of CHD repairs in adults continues to increase, these operations will be performed in a wider variety of institutions and systems. Unfortunately, not all of these centers will have an optimal environment for correcting CHD in adults. This type of surgery is best accomplished in a facility specifically designed for treating adults with CHD. Optimal care of these patients is provided by cardiologists who are trained and experienced in pediatric and adult cardiology, by surgeons who are trained and experienced in treating CHD, and by anesthesiologists who are experienced in caring for adults with CHD. Whatever the setting, cardiac anesthesiologists involved in these cases must be thoroughly aware of the anesthetic implications for the unique pathophysiology of each patient, and they must not rely on their "usual" expectations of either true pediatric CHD or acquired adult heart disease. PMID- 14562572 TI - Gender and cardiac surgery. AB - The increased operative mortality and morbidity of women compared with men undergoing CABG surgery results from multiple differences in presentation, preoperative risk profile, and surgical factors. Investigators have found consistently that women present with a different preoperative risk profile than do men. Women more commonly have factors associated with increased short- and long-term mortality, such as less frequent use of IMA grafts. Differences in study design and patient population may contribute to variability in short- and long-term mortality among the various studies. The lack of representation of women in older clinical trials has hindered our understanding of the management of CAD in women; this situation must be remedied in future studies, [95]. Known physiologic and anatomic differences must be evaluated for their effects on outcomes. Further studies are needed to evaluate gender-related differences in autonomic responses to acute coronary occlusion, complications related to cardiopulmonary bypass, susceptibility to abnormalities in coagulation, and other factors that might account for discrepant outcomes in men versus women undergoing CABG [96]. Beyond these factors, specific pharmacologic and therapeutic considerations, such as the role of estrogen replacement therapy, need to be clarified. As further knowledge accumulates, it is hoped that gender-specific risk factors can be mitigated and protective factors exploited, thereby improving the outcomes for all cardiac surgery patients. PMID- 14562573 TI - Pediatric gastroenterology. Preface. PMID- 14562574 TI - Genetic disorders of the pancreas. AB - The venues opened to all by the remarkable studies of the genome are just starting to become manifest; they can now distinguish different variants of a disease; they are given the tools to better understand the pathophysiology of illness; they hope to be able to provide better treatment alternatives to our patients. The examples described in this review demonstrate the applicability of these concepts to pancreatic disorders. Researchers may be just scratching the surface at this time, but the potential is enormous. Many philosophic and ethical questions need to be answered as physicians move along: Should all family members of an index case be screened? Who should pay for testing? Who should get results? But, without the participation of so many patients, their family members, and numerous volunteers, researchers would not have witnessed the bridging of so many gaps as they have so far. All of us may now look forward to the application of this incredible knowledge to the therapeutic solutions so eagerly awaited. PMID- 14562576 TI - The genetics of Hirschsprung disease. AB - Understanding the genetics of Hirschsprung disease will naturally expand our understanding of other neurocristopathies, the enteric nervous system, and autonomic system biology. As other disorders of gastrointestinal motility are investigated, genetics may resolve certain clinical questions. For example, isolated hypoganglionosis without aganglionosis has been reported as a primary cause of intestinal pseudo-obstruction. Is such hypoganglionosis merely a forme fruste of Hirschsprung disease, or a result from an entirely different pathogenetic mechanism? Can irritable bowel syndrome or severe constipation be related to specific mutations, polymorphisms, or haplotypes? How might an understanding of derangements of the ENS be translated to understanding derangements of the CNS? Clearly, we should anticipate improved prognostication, counseling, and hopefully, therapies with future genetic insights. PMID- 14562575 TI - Gastrointestinal manifestations of mitochondrial disease. AB - Although non-specific gastrointestinal and hepatic symptoms are commonly found in most mitochondrial disorders, they are among the cardinal manifestations of several primary mitochondrial diseases, such as: mitochondrial neurogastrointestinal encephalomyopathy; mitochondrial DNA depletion syndrome; Alpers syndrome; and Pearson syndrome. Management of these heterogeneous disorders includes the empiric supplementation with various "mitochondrial cocktails," supportive therapies, and avoidance of drugs and conditions known to have a detrimental effect on the respiratory chain. There is a great need for improved methods of treatment and controlled clinical trials of existing therapies. Liver transplantation is successful in acquired cases; however neuromuscular involvement in primary mitochondrial disorders should be a contraindication for liver transplantation. PMID- 14562577 TI - Disorders of bile formation and biliary transport. AB - A wide range of cholestatic liver diseases result from various primary defects in bile formation. Clinical features include jaundice, pruritus, failure to thrive, fat malabsorption, cholelithiasis, and variably progressive cirrhosis. Accurate diagnosis of these disorders is essential for determination of prognosis and selection of the most appropriate therapies. Severe genetic defects in canalicular bile acid and phospholipid excretion lead to progressive liver disease that often requires liver transplantation. Defects in bile acid biosynthesis and aminophospholipid transport may be responsive to medical or non transplant surgical approaches. PMID- 14562578 TI - Heritable disorders of the bile ducts. AB - Diseases of the bile ducts encompass a wide range of disorders. These include those disorders primarily affecting extra and intrahepatic bile ducts and those that may be classified as panbiliary. The major heritable bile duct disorders are those affecting the intrahepatic ducts, namely syndromic bile duct paucity, or Alagille syndrome, and the fibrocystic cholangiopathies autosomal recessive polycystic kidney disease/congenital hepatic fibrosis, and autosomal dominant polycystic kidney disease. This discussion focuses on heritable disorders of the bile ducts. PMID- 14562579 TI - Neonatal hemochromatosis. AB - Neonatal hemochromatosis is a rare gestational condition in which iron accumulates in the fetal tissues in a distribution like that seen in hereditary hemochromatosis. Extensive liver damage is the dominant clinical feature, with late fetal loss or early neonatal death. NH recurs within sibships at a rate higher than that predicted for simple Mendelian autosomal-recessive inheritance, possibly suggesting the role of a maternal factor. Immunomodulation during pregnancy at risk appears to lessen the severity of disease. PMID- 14562580 TI - Biliary atresia. AB - BA is a rare disease of unclear etiology; nevertheless, its impact in the field of pediatric hepatology is significant. It is the most common surgically correctable cause of neonatal cholestasis and is the most common pediatric disease referred for liver transplantation. Little progress has been made with regard to improving outcome or understanding its pathogenesis in the past decade. Fortunately, however, a national, government-sponsored collaborative endeavor has begun that will hopefully make a significant impact upon the progress of designing new treatments for BA and develop a better understanding of its pathogenesis. PMID- 14562581 TI - Overview of pediatric liver transplantation. AB - Liver transplantation is a successful and useful therapy for children with chronic or end-stage liver disease and those with a variety of extrahepatic metabolic diseases that can be corrected by liver replacement. Major developments in medical management, donor availability and procurement, operative techniques, and post-operative management all contributed to the dramatic improvement in outcome and growth of liver transplantation from its inception in 1963 [1,7,21]. The excellent long-term outcomes that have been achieved are the product of the coordinated efforts of liver transplant surgeons, specialists in pediatric hepatology and other medical areas, nurses, local practitioners, and the patients and their families. Future advances in transplant immunobiology, organ availability and infection control are likely to lead to still greater improvements in short and long-term outcome [34]. PMID- 14562582 TI - Advances in celiac disease. AB - In recent years, it has become evident that CD is much more common than previously appreciated, with a prevalence of 0.5% to 1% in Western, Arabian, and Indian populations. The disease may be present without symptoms (silent CD) or may present with extraintestinal manifestations only. Increasing awareness of the many faces of CD will increase diagnosis rate. CD patients have a cure for their disease, named the gluten-free diet, but this curative measure is very hard to adhere to. With the new insights into the pathogenesis of CD, clinicians enter an era where new treatment modalities for CD may turn into reality. PMID- 14562583 TI - Eosinophilic esophagitis. AB - Since Kelly's article appeared in press in 1995, EoE has become an increasingly recognized body. While isolated case reports suggesting EoE have been printed in the adult literature since 1977, many more reports have been published in the pediatric literature over the past 7 years. Recently, [table: see text] several excellent reviews have been written on the subject of EoE [2,25,59,60]. These reviews coupled with the past reports and increasing incidence of EoE provide a sound background for further research on the etiology, epidemiology and treatment of EoE. PMID- 14562584 TI - Inflammatory bowel disease in early childhood and adolescence: special considerations. AB - Several aspects of IBD overlap between pediatric and adult population. Those include nutritional issues, bone density, and medical and surgical therapies. Some aspects like natural course of the disease, and epidemiology and genetics are more easily examined and researched in the pediatric population. Others like pubertal and growth delay, and transition of health care are unique to pediatric patients. This article examines some of the similarities, as well as differences of IBD in these two populations. PMID- 14562585 TI - Cyclic vomiting syndrome: a brain-gut disorder. AB - Despite the "black box" surrounding CVS, the authors' understanding of this clinical entity has advanced substantially in the last decade as a result of an international interdisciplinary clinical and research effort. Although CVS is now recognized as a unique clinical entity, patients still undergo innumerable hospitalizations and diagnostic tests. Although controlled therapeutic studies are lacking, reasonably effective empiric approaches have been developed by trial and error using anti-migraine, anti-emetic, and anti-epileptic regimens. The ongoing investigations of migraine mechanisms through NMR spectroscopy, mitochondrial DNA mutations and cellular energetics, corticotropin-releasing factor and gastric motility, and brainstem regulation of autonomic function may lead to breakthroughs in the understanding of and new therapies for CVS in the next decade. PMID- 14562586 TI - Neonatal hypothermia on admission to a special care unit in Dar-es-Salaam, Tanzania: a cause for concern. AB - OBJECTIVE: To determine the prevalence and risk factors for hypothermia among neonates on admission to the Neonatal Care Unit. Hypothermia in newborn babies is a problem in tropical countries despite warm environmental conditions and it contributes to a high neonatal morbidity and mortality. METHODOLOGY: A study was undertaken to determine the prevalence of hypothermia and its association with early neonatal outcome among neonates admitted to the Neonatal Care Unit of Muhimbili Medical Centre. At admission all neonates were examined and axillary temperature recorded using a low-reading thermometer. Six-hourly temperature was taken in all infants. Those with a temperature below 36.5 degrees C were recruited as cases and those with normal temperature served as controls. These neonates were followed up for early neonatal outcome. RESULTS: Hypothermia on admission was found in 366 out of 1,632 babies (22.4%). In none of these was hypothermia recorded or reported as a reason for admission. Thirteen percent of the hypothermic neonates had severe hypothermia, with body temperature below 32 degrees C on admission. Hypothermia was significantly associated with deliveries from outside hospitals and with those who had operative or instrumental delivery in the same hospital. It was also associated with prematurity, low birth weight babies, time taken to transfer the baby and inadequate clothing after delivery. It was found that hypothermic infants had a three fold higher mortality and morbidity. These infants had a longer stay in the unit and had a higher post natal weight loss. There was no low-reading thermometer in the unit. CONCLUSION: It is concluded that there is cause for concern about hypothermia in the neonates at Muhimbili Medical Centre. Efforts should be made to sensitize and educate all levels of staff dealing with neonates, and low-reading thermometers should be part of the essential kit in the unit. PMID- 14562587 TI - HIV/AIDS knowledge and attitudes amongst pharmacists in Zimbabwe. AB - OBJECTIVE: To examine the knowledge of HIV/AIDS amongst pharmacists in Zimbabwe. DESIGN: Cross sectional survey. SETTING: National survey of pharmacists in major urban areas and final year pharmacy students at the University of Zimbabwe. SUBJECTS: 250 pharmacists and 47 pharmacy students with 126 completed questionnaires returned (42.4%). INTERVENTIONS: The questionnaire asked for demographic details of the respondent, sources of knowledge about HIV/AIDS and measured knowledge (KW), fear of contagion (FC), negative emotions (NE) and professional resistance (PR) using a Likert scale of 1 to 5. MAIN OUTCOME MEASURES: Computed scores of KW, FC, NE and PR and reported sources of knowledge on HIV/AIDS. RESULTS: All pharmacists scored highly on KW (mean [SEM] = 4.0 [0.0] with academics scoring the highest (4.2 [0.5]; n = 7) and private hospital pharmacists the lowest (3.7 [0.2]; n = 5). Medical books/journals and professional colleagues were the most important sources of information. Scores for FC (2.7 [0.1]), PR (2.4 [0.1]) and NE (2.1 [0.1]) were low. Government hospital pharmacists tended to have a higher KW score than those in private hospitals (4.1 [0.3] vs 3.7 [0.2] respectively). Most of the respondents believed that HIV/AIDS had increased their workload. However, they saw a role for pharmacists in the prevention and management of HIV/AIDS but felt there were important time constraints. CONCLUSION: Pharmacists can play an important part in the strategy to manage the national HIV/AIDS epidemic, but negative attitudes towards HIV/AIDS sufferers may adversely affect their efficiency. PMID- 14562588 TI - Use of culture methods for recovery of atypical mycobacteria from stools of AIDS patients. AB - OBJECTIVE: To establish recovery rates of atypical mycobateria from stools of suspected AIDS patients using culture media. DESIGN: Laboratory evaluation of recovery rates, contamination rates, optimum exposure time and optimum concentration of alkali used for decontamination. SETTING: The study was conducted in Harare, Zimbabwe at two medical institutions: Beatrice Road Infectious Diseases Hospital (BRIDH) (a tuberculosis referral hospital) and Mashambanzou Care Unit (MCU) (a home-based care centre). SUBJECTS: A total of 386 stool specimens from suspected AIDS patients from the two health institutions plus 81 stool specimens from clinically healthy patients were collected. The number of patients from MCU was 144 (49 females, 95 males) and 242 from BRIDH (119 males, 123 females). MAIN OUTCOME MEASURE: The main goals were to determine optimum exposure time and optimum concentration of alkali used in decontamination and to identify the culture medium with the best recovery rates of atypical mycobacteria. RESULTS: Optimum recovery of atypical mycobacteria was achieved on Peizer TB medium after treating stool specimens with 4% sodium hydroxide for 35 minutes. In addition, the use of Kirchner's medium improved isolation rates, although with a slight increase in contamination at levels of 2.9%. CONCLUSION: A stool specimen can be used to recover atypical mycobacteria in suspected AIDS patients. Recovery is achieved using Peizer TB medium at a concentration of 4%. Varying the exposure time of the stool specimen to the decontaminating alkali and incorporating antifungal agents and antibiotics into the medium, improves recovery of atypical mycobacteria. PMID- 14562589 TI - The influence of socio-economic factors on Helicobacter pylori infection rates of students in rural Zambia. AB - OBJECTIVES: Although prevalence of disease in sub-Saharan Africa is often quite high and attracts much research, relatively little is known about less critical maladies. We examined Helicobacter pylori infected students in rural Zambia. We attempted to determine if any socio-economic or co-occurring diseases were correlated to H. pylori infection. Understanding the context in which H. pylori infections occur may increase our understanding of this organism. DESIGN: We conducted a screening survey with diagnostic tests of primary and secondary school students to determine rates of H. pylori infection. We then correlated these rates to socio-economic factors such as income and tobacco use. We also explored the correlation of H. pylori to HIV and malaria. SETTING: Zimba, Zambia. SUBJECTS: Eighty seven primary and secondary school students. MAIN OUTCOME MEASURE: Correlation of H. pylori to socio-economic factors. RESULTS: H. pylori infection was common (60.9%) and was consistent with rates found in other African countries. We found no significant correlation between H. Pylori and disease and socio-economic variables. CONCLUSION: In the studied population H. pylori infection does not appear to be correlated with the measured socio-economic or disease variables. PMID- 14562590 TI - Specialisation patterns of medical graduates, University of Malawi College of Medicine, Blantyre. Cent Afr J Med 2002;48:14-7. PMID- 14562591 TI - The Malawi College of Medicine coming of age(Central African Journal of Medicine 2002;48 (1/2):14-16). PMID- 14562592 TI - The trend of ocular surface squamous neoplasia among ocular surface tumour biopsies submitted for histology from Sekuru Kaguvi Eye Unit, Harare between 1996 and 2000. AB - OBJECTIVE: To determine the trend in the prevalence of ocular surface squamous neoplasia (OSSN) among ocular surface tumour biopsies submitted for histology from Sekuru Kaguvi Eye Unit (SKEU), Harare between January 1996 and August 2000. DESIGN: The study was a retrospective records review of patients who had biopsies of ocular surface tumours done during the study period. SETTING: National Referral Centre for eye patients (Sekuru Kaguvi Eye Unit), Parirenyatwa Hospital, Harare, Zimbabwe. SUBJECTS: A total of 1,587 records of patients who had ocular surface tumour biopsies performed were reviewed. MAIN OUTCOME MEASURE: Period prevalence of OSSN among ocular surface tumour biopsy specimens submitted for histology during the period under review. RESULTS: The median age of patients with OSSN was significantly different from the median age of patients with other conjunctival tumours: 35 years (interquartile range [IQR] 25 to 47 years) versus 51 years (IQR 25 to 60 years), p < 0.0005 respectively. Four hundred and thirty out of 615 (70%) patients with OSSN were females. The overall period prevalence of OSSN was 615 of the total of 1,587 (38.8%). The annual prevalence of OSSN among ocular surface tumour biopsy specimens was found to be on the increase and ranged from 33% (115) in 1996 and 57.9% (165) during the year 2000. The Chi squared test for trend analysis showed a significant upward linear trend (x2 = 27.4, p < 0.004). Fifty three percent of the tumours were OSSN, 42% were pterygium and 5% were pyogenic granulomas of the conjunctiva. CONCLUSION: OSSN is the commonest tumour of the ocular surface whose primary site is the conjunctiva. Its prevalence in Zimbabwe is on the increase and it has a predilection for females over males. Further studies on the natural history, treatment and patient profile would pave a way to better quality of patient care and establish its association with HIV/AIDS. PMID- 14562593 TI - Validation of a simple and cheap gelatin particle agglutination test for human immunodeficiency virus using dried blood spot samples. AB - OBJECTIVES: To validate a Gelatin Particle Agglutination (GPA) test for HIV using dried blood spots with a view to applying the test in large epidemiological studies. DESIGN: A method comparison study with standard enzyme linked immunosorbent assay (ELISA) using the Recombigen HIV-1/2 kit as the gold standard. SETTING: Blair Research Laboratory, Harare, Zimbabwe. SUBJECTS: Sera and dried blood spots samples were available from 379 women from Mbare, Harare and Mupfure, Shamva District, Zimbabwe who had participated in HIV studies conducted by Blair Research Laboratory. MAIN OUTCOME MEASURES: Results of the GPA and Recombigen HIV-1/2 ELISA using serum and dried blood spots. RESULTS: With the Recombigen HIV-1/2 ELISA as the gold standard, sensitivity and specificity of the GPA were 100% and 99.2% respectively using serum. With dried blood spots sensitivity and specificity of the GPA test were 100%. The cost of analysing one sample, based on cost of reagents and accessory materials only, was Z$300 for GPA compared to Z$1,200 for ELISA. Furthermore, hands-on time was significantly reduced with the GPA compared to ELISA. CONCLUSION: The GPA method is simple, less labour intensive and much cheaper, yet is equally sensitive when compared to standard ELISA. The high sensitivity with blood spots makes the test ideal for large-scale epidemiological studies in remote rural areas with no infrastructure for advanced diagnostic methods. PMID- 14562594 TI - Prevalence and intensity of the schistosomiasis situation along the Zimbabwean urban and peri-urban shoreline of lake Kariba. AB - OBJECTIVE: To determine the prevalence and intensity of schistosomiasis in urban and peri-urban Kariba. DESIGN: A cross sectional schistosomiasis survey involving screening people for infection and searching for intermediate host snails. SETTING: Lake Kariba Research Station, Zimbabwe. SUBJECTS: One thousand and seventy three people living in four residential areas of Kariba were screened for both Schistosoma haematobium and Schistosoma mansoni. Seventeen water contact sites along the shoreline close to the residential areas were surveyed for snails. MAIN OUTCOME MEASURES: Prevalence and intensity of both S. haematobium and S. mansoni and abundance of intermediate host snails at potential transmission sites. RESULTS: The prevalence of S. haematobium and S. mansoni were 7.2% and 2.1% respectively. More males (8.4%) were infected with S. haematobium than females (5.7%). There was no difference in prevalence of S. mansoni between males (2.2%) and females (2.0%). Most infections of S. haematobium were in the age group of 15 to 19 years while the age distribution pattern for S. mansoni was patchy. Most of the infections detected were light i.e. one to 100 eggs per gram of faeces or less than 50 eggs per 10 ml of urine. Few intermediate host snails for schistosomiais were collected and none of them were infected with human schistosomes. CONCLUSION: Prevalence and intensity of schistosomiasis were low and therefore, the disease was not considered to be a major public health problem. PMID- 14562595 TI - The rate of and indications for enucleations at Sekuru Kaguvi Eye Unit in Harare: a comparative analysis. AB - OBJECTIVES: To determine the rate of and indications for enucleation at Sekuru Kaguvi Eye Unit (SKEU) in Zimbabwe and compare the findings with those from other tertiary eye care centres in the developing world. DESIGN: Descriptive retrospective cross sectional study. SETTING: Sekuru Kaguvi Eye Unit, Parirenyatwa Hospital, Harare, Zimbabwe. SUBJECTS: Records of patients who had undergone enucleation at Sekuru Kaguvi Eye Unit, between January 1988 to December 2000. MAIN OUTCOME MEASURES: Rate and indications of enucleations done at SKEU during the study period. RESULTS: Ocular tumours (64.2%) and ocular infections (26.4%) were the leading indications for enucleation and the crude incidence rate for enucleation at SKEU was 0.17% CONCLUSION: Indications for enucleation at the SKEU in Harare are similar to those experienced by other developing countries. PMID- 14562596 TI - Highly symptom-aware children were heavily infected with urinary schistosomiasis in southern Ghana. AB - OBJECTIVES: To assess the relationship between the infection status of children and their knowledge, attitudes, beliefs, and practices (KABP) related to urinary schistosomiasis. DESIGN: Questionnaire survey. SETTING: Nine schools in eight rural communities (total population: 4,636) in Ga and South Akuapem Districts, Ghana. SUBJECTS: Four hundred and six children attending primary and secondary schools. MAIN OUTCOME MEASURE: Schistosoma haematobium infection status of children and their KABP. RESULTS: Of 354 children who responded and also submitted their urine samples, 297 (83.9%) tested positive for S. haematobium and the intensity of infection was 90 (95% CI: 74 to 110) eggs per 10 ml urine. General knowledge variables such as the knowledge of symptoms (p < 0.001), and knowledge of swimming or bathing in the river as a transmission route (p < 0.001) showed significant association for higher prevalence and intensity of infection. Treatment-seeking behaviour was not associated with the lower prevalence or intensity of infection. Practice variables such as washing clothes in the stream (p < 0.001) and fishing in the stream (p < 0.01) were significantly associated with both higher prevalence and higher intensity of infection. Children who knew of contact with river water as a transmission route reported more water contact activities (p < 0.001). CONCLUSION: This study showed that highly symptom-aware people were heavily infected, and people frequently exposed to infested water were heavily infected. Moreover, highly symptom-aware people never constituted a group whose exposure was slight. Why was awareness of disease symptoms and general knowledge of the disease not linked to low infectivity? Why didn't awareness result in avoidance of infested water sources? This report highlights the urgent need to address these important questions in future research. PMID- 14562597 TI - Vaginal misoprostol for induction of labour: a more effective agent than prostaglandin F2 alpha gel and prostaglandin E2 pessary. AB - OBJECTIVES: To compare labour outcome in women who had labour induced with PGF2 alpha gel, PGE2 vaginal pessary or misoprostol administered intravaginally or orally. STUDY DESIGN: Unmasked randomised controlled trial. SETTING: Department of Obstetrics and Gynaecology, University of Zimbabwe, Harare. SUBJECTS: Women with a singleton foetus in cephalic presentation after 37 weeks gestation admitted for induction of labour who were randomised to prostaglandin F2 alpha gel (n = 76), prostaglandin E2 pessary (n = 75) and misoprostol administered either intra-vaginally (n = 128) or orally (n = 127). MAIN OUTCOME MEASURES: Primary outcome was induction to delivery interval. Secondary outcomes included use of oxytocin during labour, mode of delivery, duration of labour, neonatal condition at delivery and maternal complications. METHODS: Four hundred and six women admitted for induction of labour with a singleton foetus in cephalic presentation after 37 weeks gestation were enrolled. To estimate the risk with induction using other agents the odds ratio and 95% confidence interval was calculated using the group that received prostaglandin F2 alpha gel as referents. RESULTS: The women were comparable for baseline characteristics. Compared to prostaglandin F2 alpha gel, the need for augmentation with oxytocin in labour was significantly reduced in women induced with prostaglandin E2 pessary (OR 0.46; 95%CI 0.23 to 0.93), vaginal misoprostol (OR 0.34; 95%CI 0.18 to 0.63) and oral misoprostol (OR 0.42; 95%CI 0.22 to 0.78). There was no difference in mode of delivery. There was a significantly reduced risk (OR 0.20; 95%CI 0.04 to 0.86) of Caesarean section (CS) for failure to progress in the vaginal misoprostol group. Labour induced with misoprostol and prostaglandin E2 pessary was significantly shorter than in prostaglandin F2 alpha gel. Vaginal misoprostol significantly shortened the induction to delivery interval. There were more admissions to the neonatal unit in the misoprostol groups. CONCLUSION: Compared to prostaglandin F2 alpha gel, misoprostol and prostaglandin E2 pessary had reduced need for oxytocin and a shorter duration of labour. Effects of misoprostol on the foetus need further investigation before it is used as a routine agent for induction of labour. PMID- 14562598 TI - Effect of Plasmodium falciparum parasitaemia on some haematological parameters in adolescent and adult Nigerian HbAA and HbAS blood genotypes. AB - OBJECTIVE: To compare the susceptibility of HbAA and HbAS blood genotypes to Plasmodium falciparum in human subjects. DESIGN: Prospective Cross Sectional Study. SETTINGS: Department of Biochemistry College of Medical Sciences, University of Calabar, Cross Rivers State, Nigeria. SUBJECTS: A total of 120 individuals comprising 60 subjects (41 males and 19 females) and 60 patients (33 males and 27 females) aged between 15 and 30 years were enlisted in this study. MAIN OUTCOME MEASURES: The susceptibility of the HbAA and HbAS blood genotypes to Plasmodium falciparum in human subjects was based on the frequency of occurrence of parasitaemia and the accompanying haematological changes in each group. RESULTS: Among a sample population of 120 subjects and patients, 86 were HbAA and 34 were HbAS representing 72% and 28% of the sample population. Parasites were detected in the red cells of 15 out of the 34 HbAS subjects i.e. 44% of the group or 13% of the sample population. Compared with the control group, the parasitaemic HbAA group had significantly lower PCV and Hb, and significantly higher serum iron, TIBC and transferrin, but a similar percentage transferrin saturation. All the differences were highly significant (p < 0.001). In the parasitaemic HbAS group, PCV and percentage transferrin saturation were slightly lower while TIBC and transferrin were slightly higher when compared with the control. These differences were not significant. CONCLUSION: The greater susceptibility of HbAA individuals to Plasmodium falciparum malaria and the enhanced severity of an attack in this group maybe due to low red cell membrane resistance to the invading parasite and a non-hypoxic environment within the red cell which enhances its development. PMID- 14562599 TI - Levels and risk factors for mortality in infants with birth weights between 500 and 1,800 grams in a developing country: a hospital based study. AB - OBJECTIVE: To determine levels of mortality and risk factors for mortality in infants born with birth weights below 1,800 gms. DESIGN: Prospective descriptive study. SETTING: Harare Central Hospital in Zimbabwe. SUBJECTS: All infants born and admitted to Harare Hospital Neonatal Unit between January and May 2000, with birth weight between 500 gms and 1,800 gms. STUDY FACTORS: Mothers' age, parity, booking status, mode of delivery, infants' sex, birth weight, use of intensive care, outcome in hospital, age at death and age at discharge from hospital. RESULTS: Four hundred and ninety infants were studied. In hospital, the fatality rate was 39.4%. Only 49.4% of mothers had received antenatal care. The median birth weight of those who died was 1,077 gms (Q1 = 500, Q3 = 1,357) while that of the discharged infants was 1,530 gms (Q1 = 850, Q3 = 1690). Risk factors for mortality were birth weight less than 1,500 gms compared to 1,500 gms or more. Odds Ratio (OR) 7.53 (95% CI = 4.66 to 12.23), breech delivery compared to vaginal delivery, OR 2.40 (95% CI = 1.28 to 4.52) and lack of antenatal care OR 1.59 (95% CI = 1.08 to 2.33). Parity, sex of infant and receiving intensive neonatal care were not significantly associated with mortality. CONCLUSION: Strategies to reduce mortality in these infants should include better access to early, high quality obstetric care and avoidance of breech delivery in preterm infants. Identification of avoidable factors leading to preterm delivery is critical. PMID- 14562600 TI - Determination of normal ranges for serum C3 and C4 levels in an adult Zimbabwean population. AB - OBJECTIVE: To determine reference levels of serum complement proteins (C3 and C4) by analyzing the concentrations of this serum component in blood samples drawn from otherwise healthy adult Zimbabweans. DESIGN AND SETTING: This was a cross sectional study carried out at the University of Zimbabwe College of Medicine and National Blood Transfusion Services, Harare, Zimbabwe. SUBJECTS: Ninety five ethnically heterogenous apparently healthy blood donors aged between 18 and 60 years were enrolled in the study. These included 50 males and 45 females. MAIN OUTCOME MEASURES: Serum C3 and C4 levels were determined using an immuno turbidimetric method. RESULTS: Serum levels of complement (C3) ranged from 66 to 185 mg/dl. The respective levels of C4 were 15 to 52 mg/dl. These values varied significantly with age. Serum levels of C3 in patients aged 18 to 50 years were 63 to 192 mg/dl; they were lower (80 to 124 mg/dl) in patients in the 51 to 60 year old age bracket. C4 levels were lower (12 to 49 mg/dl) in the younger adults (18 to 40 years) than in those aged 41 to 60 years (24 to 53 mg/dl). The levels of both C3 and C4 did not vary with gender. CONCLUSION: This study is probably the first to establish local reference ranges for C3 and C4 in Zimbabwe. These results should find useful clinical and research application. PMID- 14562601 TI - Seven essential oils inhibit Anopheles arabiensis mosquito biting. PMID- 14562602 TI - Reduction of postoperative adhesions by N,O-carboxymethylchitosan and spermine NONOate in rats. AB - BACKGROUND: Postsurgical adhesions can occur following virtually all types of surgery, resulting in serious clinical complications. Therefore, prevention of adhesions is an important goal of surgical practice. A rat uterine horn model was used to investigate the efficacy of N,O-carboxymethylchitosan (NOCC) and spermine NONOate (SPER/NO) alone and in combination in preventing adhesion formation. METHODS: Sixty Wistar albino rats underwent bilateral uterine horn injury with a unipolar cautery. Study groups were as follows: (i) control group, no adjuvant therapy; and those with adjuvant applied, (ii) normal saline group, 2 ml of normal saline was given; (iii) NOCC group, 2 ml of 2% NOCC gel was given; (iv) SPER/NO group, 2 ml of SPER/NO (0.5 mg/ml) was given, and (v) NOCC plus SPER/NO group, 2 ml of 2% NOCC gel including SPER/NO (0.5 mg/ml) was given. After 14 days, all animals were euthanatized, and a standard adhesion scoring system including extent and severity scores was applied by a blinded examiner. RESULTS: The extent score in NOCC plus SPER/NO group was significantly lower than those of control and normal saline groups (p < 0.05). The extent score in NOCC group was significantly lower than that of normal saline group (p < 0.05). The extent score in NOCC plus SPER/NO group was significantly lower than that of SPER/NO group (p < 0.05). The severity score was significantly lower in NOCC plus SPER/NO and NOCC groups than that of control group (p < 0.05). The severity score was significantly lower in NOCC plus SPER/NO group than that of SPER/NO group (p < 0.05). CONCLUSIONS: Postoperative administration of NOCC gel and SPER/NO alone and especially in combination to the site of peritoneal injury reduces the formation of adhesions in the rat uterine horn model. PMID- 14562603 TI - Insertional mutation in the intron 1 of Unc5h3 gene induces ataxic, lean and hyperactive phenotype in mice. AB - Mice carrying a mutation in the first intron of Unc5h3 were accidentally produced by transgenic insertion and characterized for their homozygous mutant phenotypes. Morphological and histological analysis revealed cerebellar and midbrain abnormalities, which are similar to the previously reported phenotypes of the Unc5h3 mutant. Behavioral analysis showed higher ambulatory activity and circling, and defects in habituation in a novel environment. Their body weights were 10-30% less than wildtype mice from 2-3 weeks of age to 22 months possibly due to reduced accumulation of adipose tissues. The transgenic insertion site was identified and mapped to the intron 1 of Unc5h3 gene with approximately 50 kb deletion of the intron sequence. This intronic mutation interfered with the mRNA expression of the Unc5h3 gene not in testes, but in many tissues including the brain, implying that this intronic region may play a role in regulating tissue specific transcription of Unc5h3. PMID- 14562604 TI - Progressive development of pulmonary hypertension leading to right ventricular hypertrophy assessed by echocardiography in rats. AB - The present study aimed to evaluate the development of pulmonary hypertension by serial echocardiography, including measurements of pulmonary artery (PA) flow velocities, and correlate echocardiographic indices with pathological findings in rats administered monocrotaline (MCT). MCT (60 mg/kg body weight) or physiologic saline was administered to a total of 9 male Wistar rats at the age of 4 weeks (MCT group: n = 4, control group: n = 5, respectively). Echocardiography was performed serially until the age of 8 weeks. The ratio of right ventricular (RV) outflow tract dimensions to aortic dimensions increased progressively in the MCT group and became significantly greater than that of the control group after the age of 6 weeks. Peak PA velocity (Peak V) in the MCT group was significantly less than that of the control group at the ages of 7 and 8 weeks. The ratio of acceleration time to ejection time (AT/ET) in PA flow waveforms declined progressively and was significantly less than that of the control group after the age of 6 weeks. The ratio of RV weight to body weight (RVW/BW) in the MCT group was significantly greater than that of the control group. Both AT/ET ratio and Peak V were significantly inversely correlated with RVW/BW ratio. Furthermore, these echocardiographic findings were also significantly inversely correlated with the mean cross-sectional RV myocyte area. In conclusion, the progressive development of pulmonary hypertension leading to RV hypertrophy can be evaluated appropriately by echocardiography including PA flow Doppler indices in rats. PMID- 14562605 TI - Establishment and characterization of the Komeda diabetes-prone rat as a segregating inbred strain. AB - The Komeda diabetes-prone (KDP) rat is a spontaneous animal model of human autoimmune type 1 diabetes. By positional cloning of the non-MHC major susceptibility locus lddm/kdp1, we recently identified a nonsense mutation in Cblb and also found that lymphocytes of KDP rats infiltrate into various tissues, indicating autoimmunity. The maintenance and production of KDP rats has been a critical problem owing to the poor reproductive ability of diabetic animals. To solve the problem, we here established the KDP rat as a segregating inbred strain. We first identified animals that were heterozygous at the lddm/kdp1 region in a breeding colony of KDP rats. The heterozygous region spans at least from D11Yok1 to Cblb on rat chromosome 11. By mating between the heterozygous rats, we obtained homozygotes, heterozygotes and wild-types with the expected ratio of 1:2:1 and found that only the homozygotes developed diabetes, suggesting that these genotypes represent those of lddm/kdp1. We then tried to maintain KDP rats by mating between the heterozygotes, which resulted in a segregating inbred strain. Within 210 d of age, about 80% of lddm/kdp1 homozygotes developed diabetes with severe insulitis, while neither heterozygotes nor wild-types developed diabetes. The phenotypic characteristics of the homozygotes are the same as those of progeny of diabetic parents in the original KDP rats. The segregating inbred KDP rat strain described here would serve as a useful animal model for autoimmune diseases, including type 1 diabetes. PMID- 14562606 TI - A congenic strain (F344.OLETF-Imfm) displays the existence of intramuscular fat accumulation QTL on rat chromosome 1. AB - A genomic region between D1Wox8 and D1Rat90 on rat chromosome 1 was previously shown to be linked to intramuscular fat accumulation by quantitative trait locus (QTL) analysis using a F2 population derived from the Otsuka Long-Evans Tokushima Fatty (OLETF) rat, which exhibits an increase in the levels of intramuscular fat content in Musculus longissimus, and the F344 rat. There exist two regions showing major and minor lod peaks for linkage to intramuscular fat accumulation, in the chromosomal region. We constructed a congenic strain introgressing the OLETF allele on the minor but not the major lod peak region in the F344 rat strain. The congenic strain had higher levels of intramuscular fat content in Musculus longissimus than the inbred partner F344 rat, thereby proving the existence of a QTL, designated Imfm (for Intramuscular fat-minor), responsible for the intramuscular fat accumulation in the congenic region of the minor lod peak region of about 10 cM. The F344.OLETF-Imfm congenic strain might provide a refined tool for the analysis of the gene causing intramuscular fat accumulation. PMID- 14562607 TI - Possible role of genetic factor(s) on age-related increase of peripheral CD4+CD8+ double positive T cells in cynomolgus monkeys. AB - Mature TCR alpha beta T cells in peripheral blood are generally classified into either CD4 single positive (sp) T cells or CD8sp T cells. Several studies demonstrated that considerable amounts of CD4+CD8+ double positive (DP) T cells exist in peripheral blood of human and several animals. In particular, we previously reported that peripheral DP T cells increase in an age-related manner in cynomolgus monkeys (Macaca fascicularis), but the finding that DP T cells in some aged monkeys were maintained at a low proportion (under 5%), suggests that the increase in peripheral DP T cells might be genetically controlled in cynomolgus monkeys. To test this hypothesis, 24 families were randomly selected and used in a formal genetic analysis of the proportion of DP T cells. Parents and offspring in selected families were classified into DP-High and DP-Low groups based on a 5% cutoff level of DP T cells. The cutoff value was set by analysis of the distribution of the proportion of DP T cells. Nine out of 13 offspring (69.2%) with DP-High x DP-High parents belonged to the DP-High group, whereas three out of nine offspring (33.3%) belonged to DP-High group in the case of DP High x DP-Low mating pairs. No offspring (0%) of two offspring with DP-Low x DP Low parents belonged to the DP-High group. In addition, heritability (h2: narrow sense) obtained from the regression coefficient of offspring on mid-parent values was 0.54 +/- 0.19. Both findings suggest that increases in DP T cells in cynomolgus monkeys may be genetically controlled. PMID- 14562608 TI - Protective effects of probucol treatment on pancreatic beta-cell function of SZ induced diabetic APA hamsters. AB - To clarify whether oxidative stress is involved in the pathogenesis of islet lesions of diabetic animals, the effects of probucol (PB), an antioxidant and anti-hyperlipidemia agent, on the islets in streptozotocin (SZ)-induced diabetic APA hamsters in the acute and chronic phases of diabetes were examined. The control (CB group) and diabetic (SZ group) hamsters were treated with PB (1% in the diet) for 4 weeks from several days after SZ injection as the acute diabetic group, or 8 weeks from 6 weeks after SZ injection as the chronic diabetic group. Glucose tolerance test revealed that PB treatment decreased the high serum glucose level after glucose injection in the diabetic APA hamsters in the acute diabetic phase. Immunohistochemistry revealed that PB treatment significantly increased the percentage of the insulin positive area in the diabetic hamsters pancreata in both the acute and chronic phases. In addition, 4-hydroxy-2-nonenal (4HNE; an oxidative stress marker) positive cells were slightly reduced by PB treatment in the acute diabetic phase. Double-immunostaining for insulin and PCNA (proliferating cell nuclear antigen) revealed that elevation of the percentage of insulin and PCNA double-positive cells against insulin-positive cells was seen in the islets of PB-treated diabetic hamsters, but the difference was not significant compared with untreated diabetic hamsters (p = 0.07). In semi quantitative RT-PCR, the expression of two genes, Reg (Regenerating gene) and INGAP (islet neogenesis associated protein), in the diabetic APA hamsters was significantly increased compared to the control groups in both diabetic phases. PB treatment significantly reduced Reg expression in the chronic diabetic phase. These data suggest that PB treatment in SZ-injected diabetic hamsters partially restored beta-cell function through acting as an antioxidant and induced higher expression of Reg and INGAP genes in the pancreas of hamsters. PMID- 14562609 TI - Reproductive features of the Russian vole in laboratory breeding. AB - Reproductive features of newly bred Russian voles (Microtus rossiaemeridionalis) as a laboratory animal were studied. This species is a copulatory ovulator, and most couples copulated 6 to 16 h after pairing. The gestation period varied from 18 to 22 days (mean +/- SD: 20.6 +/- 0.9, n = 72), and the average litter size was 4.6 +/- 1.9 (n = 125). Compared with the litter size at the first parturition (3.6 +/- 1.6, n = 72), the litter size in the subsequent parturitions increased to 5.9 +/- 1.4 (n = 53). The animals exhibited postpartum estrus, and repeated pregnancy accompanied with suckling pups and parturition continuously in the laboratory condition unlike other vole species. In view of their complex stomach and good proliferation, the Russian voles were evaluated as a good laboratory animal, especially as a model animal for ruminant studies. PMID- 14562610 TI - The hemodynamic effects of acute myocardial ischemia and reperfusion in Clawn miniature pigs. AB - Acute myocardial ischemia was induced by occluding the LAD in Clawn miniature pigs. Eight pigs (group 1) were subjected to 6 h ischemia and nine pigs (group 2) were subjected to 20 min ischemia, followed by reperfusion for 340 min. Three animals of the group 1 died due to ventricular fibrillation after occlusion and in group 2, four animals died due to the arrhythmia after reperfusion. Though the ischemic area of group 2 (15.6% of the ventricle) was narrower than that of group 1 (21.7%), the survival rate was lower. We supposed that ischemia-reperfusion injuries were strongly connected with the hemodynamics of group 2. Clawn miniature pigs are useful experimental animals for myocardial ischemic researches. PMID- 14562612 TI - A simple method for sutureless gastrointestinal anastomosis in rat. AB - A simple tubing stent was attempted to test a model for sutureless gastrointestinal anastomosis in 6 male rats at the age of 15 weeks. In the 3rd and 4th weeks after the operation, X-ray examination demonstrated that the gastrointestinal passage in the anastomotic site was quite satisfactory. There was no incidence of anastomotic leakage. In the 6th week after the operation, there were no macroscopic or microscopic ruptures, nor were there any obstructions at the anastomotic site. This simple sutureless method was effective at preparing anastomosis in the gastrointestinal tract in the rat and could be applied to other small experimental animals. PMID- 14562611 TI - Spontaneous T-cell-rich B-cell lymphoma in a cynomolgus monkey (Macaca fascicularis). AB - A spontaneous T-cell-rich B-cell lymphoma (TCRBCL) occurred as a subcutaneous mass in the buccal region and enlarged submandibular lymph node in a 6-year-old female cynomolgus monkey (Macaca fascicularis). The constituent cells were examined by histology, immunohistochemistry and the double labeled immunofluorescence method (dl-IF). Further, in situ hybridization (ISH) was employed to detect the gene expression of Epstein Barr virus (EBV). Histologically, the mass was comprised mainly of neoplastic large lymphoid cells and reactive small mononuclear cells. Immunohistochemically, the neoplastic large lymphoid cells were positive for CD20, CD79 alpha, MHC class II, and either IgG, IgM, or IgA. Polyclonal Ig production by the neoplastic large lymphoid cells was demonstrated by dl-IF, although IgG-positive ones predominated in number. On the other hand, most of the small mononuclear cells were positive for CD3 and were regarded as reactive T lymphocytes, while the remaining cells appeared to be histocytes or reactive B-cells. Transcripts of EBV gene were not demonstrated in these neoplastic or reactive cells by ISH. This is the first reported case of spontaneous TCRBCL in the cynomolgus monkey. PMID- 14562613 TI - A new method of inhalation anesthesia with nasopharyngeal insufflation in rat experiment. AB - We have established a new method of anesthesia with nasopharyngeal insufflation for intraoral procedure in rodents. Twelve male Wistar rats weighing 330-390 g were used in this study. Insertion of a feeding tube 1.0 mm in diameter coated with 2% xylocaine jelly was inserted into the nasal cavity approximately 25 mm from the naris, and anesthetization with mixed gas of 100% oxygen with 3-4% enflurane at 0.25-0.5 l/min flow rate was achieved. Using this anesthetic method, a chronic experiment comprising 1-h/day experimental procedure was carried out for 14 days. This method enabled, 1) simple and safe operation of the induction, emergence and anesthetic depth, 2) experimental procedures on the dental/oral region, 3) avoidance of the dyspnea and tachypnea, and 4) avoidance of cumulative effects in daily anesthesia. PMID- 14562614 TI - Production of age-related DNA strand breakage in brain cells of senescence accelerated prone (SAMP1) mouse. AB - Amounts of DNA strand breaks were estimated by the proportion of cells without tails (PCWT) and the average lengths of tail momentum (ALTM) in comet images of tissue cells of senescence-accelerated prone (SAMP1) mouse and senescence accelerated resistant (SAMR1) mouse. The PCWT and ALTM of brain cells from SAMR1 were unchanged from 4 to 15 months of age. In the case of SAMP1 brain cells, the PCWT decreased and the ALTM increased in an age-related manner from 8 to 15 months of age. In the cases of liver and kidney, the PCWT and the ALTM of both SAMP1 and SAMR1 cells showed constant values from 4 to 15 months of ages. PMID- 14562615 TI - Validity and application of noninvasive measurement of blood pressure in hamsters. AB - The aim of this study was to examine both the validity and application of noninvasive measurement of blood pressure (BP) at the brachial artery in hamsters. There were significant correlations between noninvasive and invasive values for systolic BP (SBP), mean BP, and diastolic BP (r = 0.96, r = 0.93, r = 0.88, respectively, all p < 0.0001). Furthermore, SBP appeared to be the most accurate noninvasive BP index because both proportional and constant systematic errors were the least among the three indices. Peripheral vascular resistance calculated by combination with echocardiographic indices actually reflected the prominent alpha 1-adrenergic drive induced by beta-adrenergic blockers. In conclusion, noninvasive BP measurement at the brachial artery can provide useful information regarding the integrated cardiovascular function in hamsters. PMID- 14562617 TI - [What should be considered for animal welfare?]. PMID- 14562616 TI - Immunohistochemical analysis of protein gene product 9.5, a ubiquitin carboxyl terminal hydrolase, during placental and embryonic development in the mouse. AB - Protein gene product 9.5 (PGP9.5) is expressed at high level in the neural and neuroendocrine systems. We investigated the localization and degree of expression of PGP9.5 in the developing mouse placenta and embryo at 6.5, 10.5 and 14 days of gestation using an immunohistochemical technique. At 6.5 days of gestation PGP9.5 was detected at various levels in decidual and primary trophoblast giant cells in the placenta, and in embryonic ectodermal cells in the embryo. At 10.5 and 14 days of gestation PGP9.5 was expressed at moderate to strong levels in neurons in the embryo, but rarely in the placenta. These findings suggest that the protein may play a significant role in implantation and placental development, and differentiation of embryonic ectoderm. PMID- 14562618 TI - [Social responsibility of scientists]. PMID- 14562619 TI - [Accountability demanded by society--legal and theoretical viewpoints]. PMID- 14562620 TI - [Animal rearing that could "withstand' information disclosure: consideration toward and establishment of animal welfare]. PMID- 14562621 TI - [Examination of plans for animal experiments and information disclosure]. PMID- 14562622 TI - [Future image of animal experimentation with its details open to public]. PMID- 14562623 TI - [The role of disease models using primates in the study of P. falciparum malaria]. PMID- 14562624 TI - [Genetic analysis of host resistance to rodent malaria in mice]. PMID- 14562625 TI - [Animal model for Helicobacter pylori infection]. PMID- 14562626 TI - [Development of mice highly sensitive to human prions]. PMID- 14562627 TI - [An approach to elucidate the physiopathology by using an animal model for type C hepatitis]. PMID- 14562628 TI - New contraceptive methods. AB - OBJECTIVE: To provide an update on the current trend toward increasing use of emergency postcoital contraception, continuous (or "extended-use") oral contraceptives (OCs) and longer-acting contraceptive delivery systems. DATA SOURCES: This review is based largely upon patient care-oriented clinical experience data, including some original work submitted to the U.S. Food and Drug Administration by the primary investigators of these novel methods, supplemented, when appropriate, with basic pharmacologic and pharmacokinetic information. METHODS OF STUDY SELECTION: This review is a distillation based on an exhaustive literature search of each of the following topics: emergency postcoital contraception, continuous (extended-use) oral contraception, the weekly patch, the monthly vaginal ring and the monthly injectable. The issue of OC formulations recently begun to be marketed in this country was excluded since they have been extensively reviewed elsewhere. RESULTS: Emergency postcoital contraception is highly effective, though not as effective as available proactive methods of hormonal contraception. Of the popular regimens available, all have similar efficacy; those without estrogen are associated with less nausea and vomiting. The prophylactic administration of an antiemetic 1 hour prior to the first contraceptive dose can mitigate this adverse side effect. Continuous (or "extended" use) of OCs has become increasingly popular. This can decrease or eliminate menstruation as well as several menstruation-related complaints (e.g., dysmenorrhea, menstrual migraines and premenstrual symptoms). Despite the fact that combination OC regimens offer highly reliable contraception as well as several noncontraceptive health benefits, they are limited by the issue of daily compliance. To address this limitation, several longer-acting delivery systems were recently developed. The weekly patch, monthly vaginal ring and monthly injectable have efficacy and side effect profiles similar to those of comparable OC formulations. They offer the advantages of longer action and lower and more steady steroid levels and are free of the challenges associated with daily compliance. CONCLUSION: We need to continue to develop new and highly effective means of contraception that offer women ease of use and minimal side effects, hence leading to successful and effective use. PMID- 14562629 TI - Hysterectomy in women with cervical stenosis. Surgical indications and pathology. AB - OBJECTIVE: To evaluate indications for surgery and final pathology results in patients who underwent hysterectomy with a concurrent diagnosis of cervical stenosis. STUDY DESIGN: Retrospective chart review of 25 women who underwent hysterectomy after diagnosis of cervical stenosis. RESULTS: The average age was 59 years (range, 38-80). Indications for surgery included postmenopausal bleeding in 10 patients, previous dysplasia with inadequate Papanicolaou test follow-up in 4, recurrent high grade squamous intraepithelial lesion in 1, chronic pelvic pain in 5, acute pain in 2, dysfunctional uterine bleeding in 2 and thickened endometrial stripe in 1 patient on tamoxifen. An attempt to obtain an endometrial sample was unsuccessful in 14 patients. Twenty patients had undergone prior gynecologic surgery. The results of final cervical pathologic examination revealed severe cervical dysplasia or carcinoma in situ in 3. Final uterine pathology revealed 1 patient with uterine adenocarcinoma and 19 patients with benign pathology, including endometritis, fibroids, adenomyosis and endometrial hyperplasia. CONCLUSION: The majority of patients with cervical stenosis resulting in inadequate cancer screening and/or symptoms resulting from stenosis will have significant benign pathology (64%), cervical dysplasia (12%) or uterine cancer (4%). For these patients, hysterectomy is a reasonable option. PMID- 14562630 TI - Perioperative factors for predicting successful hysteroscopic endometrial ablation. AB - OBJECTIVE: To investigate factors affecting the success of hysteroscopic endometrial ablation in order to improve patient counseling. STUDY DESIGN: Preoperative patient characteristics (age, parity, uterine length, and presence and location of myomas) and intraoperative factors (intracavitary findings, ablation or resection, and operator) were analyzed. Forty-three women with a uterine size of < or = 10 weeks underwent hysteroscopic endometrial ablation or resection and had a median follow-up of 20 months. All were treated with depot triptorelin, 3.75 mg, 1 month prior to the procedure. Alleviation of menorrhagia and amenorrhea was classified as treatment success. Comparative analyses were made between patients with failed vs. successful procedures and with reduced bleeding vs. those with amenorrhea following surgery. RESULTS: Thirty-seven women (86%) reported a decrease in menstrual flow or amenorrhea. In 6 patients (14%), bleeding persisted or became more severe. There were no statistically significant differences between women who had successful vs. failed procedures with regard to all preoperative and intraoperative parameters analyzed. Patients with amenorrhea were significantly older as compared to women with reduced bleeding (47.5 +/- SD 5.0) vs. 44.0 +/- SD 4.1 years, P = .03. CONCLUSION: Most women with uterine size of < or = 10 weeks may expect alleviation of menorrhagia or amenorrhea after surgery. Apart from age, all other preoparative and intraoperative factors examined had no predictive value for a successful procedure. These data are valuable for proper patient counseling before hysteroscopic endometrial ablation. PMID- 14562631 TI - Papanicolaou and thin-layer cervical cytology with colposcopic biopsy control. A comparison. AB - OBJECTIVE: To compare the accuracy of conventional Papanicolaou and fluid-based, thin-layer cervical cytology. STUDY DESIGN: Cervical cytology was performed in duplicate on women who presented for cervical screening. Papanicolaou and thin layer (ThinPrep, Cytyc Corp., Boxborough, Massachusetts) cytologic samples were collected simultaneously using a split-sample method. Cytologic slides were read and reported independently. Clinical follow-up was based on the most abnormal result. Colposcopy was performed as clinically indicated, and biopsy results were compared with cytologic diagnoses. RESULTS: Three thousand samples were compared. Papanicolaou and thin-layer results were significantly different (P = .0001), with identical diagnoses in 1,844 (61%) of patients. Eighty thin-layer (2.7%) and 177 Papanicolaou (5.9%) samples were read as limited or unsatisfactory (P < .0001). The rates of atypical squamous cells of undetermined significance (ASCUS) were not statistically different (P = .06). Thin-layer cytology was read more often as low grade squamous intraepithelial lesion (P = .001) or high grade squamous intraepithelial lesion (P = .006). Colposcopy with biopsy was performed on 291 patients. With ASCUS considered an abnormal result, thin-layer cytology was more sensitive (91% vs. 85%) but had lower positive predictive value (69% vs. 74%) than Papanicolaou cytology for the presence of cervical neoplasia. CONCLUSION: Papanicolaou and thin-layer cervical cytology yielded significantly different information. Thin-layer cytology yielded significantly fewer unsatisfactory results and was more sensitive for identifying cervical intraepithelial neoplasia. PMID- 14562632 TI - Uterine incisional necrosis complicating cesarean section. AB - OBJECTIVE: To establish the clinical features of patients with necrosis of the uterine incision after cesarean delivery because of the paucity of modern reports in the literature. STUDY DESIGN: Over a 4-year period, the records of 6 patients with this complication were reviewed. RESULTS: Three patients presented within 48 72 hours of delivery with clinical features very similar to those of bowel perforation. One patient had fascial dehiscence on day 5 after delivery. Two cases presented 2 and 6 weeks after delivery with postpartum hemorrhage without peritoneal findings. Diagnosis was not made prior to surgery in any case. All patients recovered after hysterectomy. CONCLUSION: While the major risks with this uncommon but important complication are serious infectious morbidity and mortality, in our patients these risks were avoided but at the cost of hysterectomy in a group of young women of low parity. More data in the future might establish a basis for rational management, possibly including conservative surgery in selected cases. PMID- 14562633 TI - Effect of operative vaginal delivery on the outcome of permanent brachial plexus injury. AB - OBJECTIVE: To evaluate whether operative vaginal delivery worsens the extent and/or degree of permanent brachial plexus injury. STUDY DESIGN: We utilized a dataset (n = 104) of vaginal deliveries resulting in permanent brachial plexus injury that ultimately underwent litigation. We excluded patients on whom neonatal injury information was incomplete (n = 5). Patients who had an operative vaginal delivery (n = 33) were compared with those who did not (n = 66) in regard to neonatal outcome and the location and extent of neurologic injury. RESULTS: The 2 classes were similar in demographic and obstetric variables. There were no differences in rates of 5-minute Apgar scores < 7 (17.9%, or 5/28, vs. 5.2%, or 3/57, P = .1), complete neurologic injury to the brachial plexus (C5-T1) (39%, or 13/33, vs. 38%, or 25/66; P = 1.0) or avulsion of the nerve roots (44%, or 12/27, vs. 36%, or 18/50; P = .5) between those with operative or spontaneous vaginal delivery. CONCLUSION: Operative vaginal delivery did not increase the severity of impairment in a dataset of deliveries resulting in permanent brachial plexus injury. PMID- 14562634 TI - Cell wall proteins of group B Streptococcus and low incidence of neonatal disease in southern Israel. AB - OBJECTIVE: To determine the maternal group B Streptococcus (GBS) prevalence of carriage and serotype distribution and the neonatal disease incidence to formulate a policy for treatment and prevention regarding GBS diseases in southern Israel. STUDY DESIGN: A prospective study was conducted between January and October 2000. Cultures were obtained from 681 healthy, pregnant women and processed as recommended. Samples were cultured on blood-agar plates with and without added gentamicin. GBS was identified by beta-hemolysis and a positive CAMP test and confirmed by agglutination with specific antiserum. Serotyping was done by the Lancefield precipitin method using monospecific antisera to polysaccharides Ia, Ib and II-VIII and surface proteins C, R and X. RESULTS: Carriage prevalence of 12.3% and neonatal disease incidence of 0.095/1,000 live births were documented. Surface proteins C and R were found in 85.7% of positive cases. Serotypes Ia (17.8%), Ib (10.7%), II (27.4%), III (20.2%) and V (14.3%) were distributed as previously reported from developed countries. CONCLUSION: Developing a pentavalent vaccine based on serotypes Ia, Ib, II, III and V in conjugation to a GBS cell wall protein transporter, such as C or R, has theoretical advantages in the southern Israeli population over vaccines that use foreign proteins. PMID- 14562635 TI - Total colpocleisis for severe pelvic organ prolapse. AB - OBJECTIVE: To describe the complications and efficacy of total colpoclesis for severe pelvic organ prolapse. STUDY DESIGN: In this descriptive study, we analyzed the complications and efficacy of total colpoclesis in 41 consecutive women with pelvic organ prolapse at Temple University Hospital between November 1994 and June 2001. RESULTS: The mean followup period was 28.7 months (range, 5 65). Vaginal hysterectomy was performed in 12 patients. All but 10 patients underwent concomitant modified paravaginal cystourethropexy. Modified perineorraphy was standard in every case. There was no recurrence of prolapse and a very low complication rate. The cure rate for urinary incontinence was 53.1% in 32 patients with preoperative urinary incontinence. CONCLUSION: Total colpocleisis combined with site-specific reconstructive surgery is an effective and safe treatment of severe pelvic organ prolapse in high-risk, elderly women. PMID- 14562636 TI - Effects of antioxidant treatment in oligozoospermic and asthenozoospermic men. AB - OBJECTIVE: To evaluate the effect of sairei-to, an herbal medicine, as an antioxidant in oligozoospermia and asthenozoospermia (nonnormozoospermia). STUDY DESIGN: Forty-seven nonnormozoospermic and 16 normozoospermic men were the subjects of this prospective clinical study. After sairei-to (9.0 g/d) was administered daily to the 2 groups for 3 months, sperm parameters, serum hormones and superoxide dismutase (SOD) activity in the serum and the seminal plasma was analyzed. The testicular artery was also assessed. RESULTS: After therapy, serum hormones and SOD activity did not change significantly in either group. Although men with normozoospermia did not undergo a significant change in sperm conditions or testicular artery flow, total sperm concentration (17.1 +/- 20.0 versus 28.7 +/- 35.5 x 10(6)/mL, P = .02) and sperm motility (30.1% +/- 21.6 versus 45.8% +/- 24.4, P < .0001) were significantly increased, and the pulsatility index of the testicular artery (2.03 +/- 0.84 versus 1.64 +/- 0.48, P = .04) was significantly decreased in nonnormozoospermia. CONCLUSION: Treatment with the herbal antioxidant sairei-to improves sperm condition and testicular artery flow in nonnormozoospermia. PMID- 14562637 TI - Consequences of nonindicated preterm delivery in singleton gestations. AB - OBJECTIVE: To determine the neonatal and economic consequences of nonindicated preterm delivery in singleton gestations. STUDY DESIGN: From a database of women with high-risk pregnancies enrolled for outpatient nursing services between October 1995 and February 2000, singleton gestations with induced labor or scheduled cesarean delivery and a gestational age at delivery of 34-36 weeks were identified. Excluded were women with preterm premature rupture of the membranes or medically indicated delivery. We compared infant neonatal intensive care unit (NICU) admission rates and ventilator use for consecutive weeks and applied a cost model to determine potential savings of delaying delivery. RESULTS: A total of 1,538 pregnancies were analyzed. Risk of NICU admission declined significantly with each advancing week (by > 50%, P <.05). NICU length of stay and total nursery costs decreased significantly between weeks 35 and 36 and weeks 34 and 35. Need for ventilatory assistance increased significantly for infants admitted to the NICU between weeks 34 and 35. The incidence of respiratory distress syndrome decreased 49% between 35 and 36 weeks. CONCLUSION: Prolonging gestation 1 week beyond weeks 34 and 35 has a significant impact on improving neonatal outcome and decreasing associated costs. These factors should be considered when electing to deliver at 34 and 35 weeks. PMID- 14562638 TI - Correlation between urodynamic test results, perineal ultrasound and degree of stress urinary incontinence. AB - OBJECTIVE: To evaluate whether the results of urodynamic tests and the perineal ultrasound were different between grade 1 and 2 stress urinary incontinence. STUDY DESIGN: Forty premenopausal women with a diagnosis of stress urinary incontinence according to urodynamic tests were enrolled in this study. Stress urinary incontinence was defined as urine leakage during stress without detrusor contraction. Twenty patients had grade 1 and 20 had grade 2 stress urinary incontinence. We compared the parameters of uroflowmetry, filling cystometry, urethral pressure profile, dynamic urethral function test, perineal ultrasound and stress urethral axis between grade 1 and 2 levels of stress urinary incontinence. RESULTS: There were no significant differences in age, parity or body mass index between the grade 1 and 2 patients. Uroflowmetry results showed that there were no significant differences in maximal flow rate, average flow rate, voided volume or residual urine between grade 1 and 2. As to cystometry results, only the first desire to void was significantly increased in grade 2 over 1 (304 +/- 113.65 vs. 194 +/- 48.24 [mL], P = .04). There were no significant differences in any of the urethral pressure profile parameters. In the dynamic test, the Valsalva leak point pressure and cough leak point pressure were not significantly different between the 2 groups. There were no significant differences in perineal ultrasound parameters or the stress urethral axis. CONCLUSION: Most stress urinary incontinence-related parameters showed no difference between the grade 1 and 2, and no urodynamic or ultrasonographic evidence for a difference between the grade 1 and 2 was demonstrated. PMID- 14562639 TI - Obstetric and gynecologic complications in women with Marfan syndrome. AB - OBJECTIVE: To analyze the maternal and fetal outcomes of pregnancy and gynecologic problems in women with Marfan syndrome. STUDY DESIGN: The outcomes of 14 pregnancies in 4 women with Marfan syndrome were prospectively observed between January 1988 and December 2000. The cardiovascular and obstetric complications were analyzed. During pregnancy all the patients were carefully monitored with serial echocardiography and close attention to symptoms. RESULTS: Of the 14 pregnancies, 5 (35.7%) ended in abortion, and 3 of them occurred in the early second trimester due to cervical incompetence. Premature onset of labor occurred in 2 pregnancies at 31 and 34 weeks. Postpartum hemorrhage complicated 3 deliveries, and inversion of the uterus occurred in 1 patient. Significant cardiovascular complications occurred in 2 patients, who required surgical correction of the aortic aneurysm and replacement of the aortic valve. In one patient the operation was performed within hours of vaginal delivery, and the other patient underwent surgery 8 weeks postpartum. No maternal death occurred in the study. One infant in the series was diagnosed as having Marfan syndrome. A premature infant delivered at 31 weeks died on the second day of life. CONCLUSION: Women with Marfan syndrome are at high risk of aortic dissection in pregnancy even in the absence of preconceptional aortic root dilatation. Obstetric complications in patients with this condition have been underreported in the past. Women with aortic root dilatation of < 40 mm usually tolerate pregnancy well, with good maternal and fetal outcomes. Women with Marfan syndrome should be counseled regarding the risk of pregnancy to both mother and fetus. Patients who have cardiac decompensation or aortic dilatation > 40 mm are advised to avoid pregnancy. PMID- 14562640 TI - Unkei-to for correcting luteal phase defects. AB - OBJECTIVE: To assess the clinical efficacy of Unkei-to in women with luteal phase defects. STUDY DESIGN: A study of 103 consecutive subjects with luteal phase defects treated with unkei-to (7.5 g/d) and 94 control subjects was conducted. A venous blood sample was taken, and plasma gonadotropins, 17 beta-estradiol and progesterone levels were measured. Dominant follicle size and endometrial thickness were assessed in the late follicular phase. RESULTS: Unkei-to significantly decreased plasma luteinizing hormone and significantly increased 17 beta-estradiol concentration in the follicular phase and significantly increased progesterone (66.7%) in the midluteal phase (P < .05). Significant development of the dominant follicle (P < .01) and endometrium (P < .001) was also observed in patients treated with unkei-to. We observed significant prolongation of the luteal phase (43%, P < .001) as measured by basal body temperature. There was a 79.6% rate of correction of luteal phase defects in patients treated with unkei to. Furthermore, the pregnancy rate in patients desiring pregnancy was 49.5% (41/83). CONCLUSION: Administration of unkei-to may normalize the gonadotropin ovarian endocrine system and/or their interaction in the follicular phase and may improve the endocrine environment in the luteal phase in patients with luteal phase defects. PMID- 14562641 TI - Association of an A allele for interleukin-10 -627 gene promoter polymorphism with higher susceptibility to endometriosis. AB - OBJECTIVE: To investigate whether interleukin-10 (IL-10) -627 gene promoter polymorphism could be used as a marker of susceptibility to endometriosis. STUDY DESIGN: Women were divided into 2 groups, endometriosis (n = 130) and nonendometriosis (n = 133). Polymorphisms for IL-10 -627 gene promoter were amplified by polymerase chain reaction and detected after restriction enzyme digestion. Genotypes and allelic frequencies in both groups were compared. RESULTS: Genotype proportions of different IL-10 gene polymorphisms in both groups were not significantly different. Proportions of A homozygote/heterozygote/T homozygote for IL-10 gene polymorphisms in both groups were 50%/40%/10% and 42.1%/38.4%/19.5%. In contrast, allele frequencies for IL-10 gene polymorphism between both groups were significantly different. Alleles A and C for IL-10 gene promoter polymorphism in the groups were 70%/30% and 61.3%/38.7%. A allele was associated with higher susceptibility to endometriosis. CONCLUSION: An association between endometriosis and IL-10 gene promoter polymorphism exists, and the IL-10 -627 A allele is related to a higher susceptibility to endometriosis. PMID- 14562642 TI - Single umbilical artery and umbilical cord torsion leading to fetal death. A case report. AB - BACKGROUND: Intrauterine fetal death is a complication that cannot often be predicted by standard obstetric management. Cord accident may be responsible for about 5% of cases. Umbilical cord torsion is an extremely rare cause of intrauterine fetal death. CASE: An 18-year-old, nulliparous woman presented with a complaint of decreased fetal movement at 38 weeks' gestation. Intrauterine fetal death was diagnosed on ultrasound. The pathologic examination revealed umbilical cord torsion and confirmed a single umbilical artery that was diagnosed on ultrasound. CONCLUSION: Umbilical cord torsion that leads to intrauterine fetal death is extremely rare. A pregnancy with a single umbilical artery may need fetal monitoring during the third trimester. PMID- 14562643 TI - Cavernous hemangioma of the cervix with intractable bleeding. A case report. AB - BACKGROUND: Cavernous hemangioma of the cervix is rare and may cause serious bleeding. CASE: A 33-year-old woman developed intractable cervical bleeding following pregnancy termination. Bleeding persisted despite curettage and suturing, and ultimately required hysterectomy. Cavernous hemangioma was confirmed histologically. CONCLUSION: This condition is a potential cause of uncontrollable cervical bleeding. PMID- 14562644 TI - Spontaneous rupture of a first-trimester gravid uterus in a woman exposed to diethylstilbestrol in utero. A case report. AB - BACKGROUND: Poor reproductive outcome was well documented in several studies of women exposed to diethylstilbestrol in utero. Spontaneous rupture of an unscarred uterus is rare and very uncommon in the first trimester of pregnancy. CASE: Spontaneous rupture of the uterus was diagnosed in a 28-year-old nullipara who developed acute abdominal pain at 12 weeks' gestation. She was known to have been exposed to diethylstilbestrol in utero. Laparotomy revealed the rupture in the anterior fundal area of the uterus. Both tubes were normal. CONCLUSION: Several spontaneous ruptures have been described, but this is the first case of first trimester spontaneous rupture of an unscarred uterus in a diethylstilbestrol exposed woman. PMID- 14562645 TI - Concomitant abdominal and intrauterine pregnancy after in vitro fertilization in a woman with bilateral salpingectomy. A case report. AB - BACKGROUND: Abdominal pregnancy is a rare event, and the concomitant presence of an intrauterine pregnancy is very exceptional. CASE: A case of concomitant abdominal and intrauterine pregnancy following in vitro fertilization and embryo transfer (IVF-ET) occurred in a woman with bilateral salpingectomy. The abdominal pregnancy was successfully treated surgically, with preservation of the intrauterine pregnancy. CONCLUSION: A careful ultrasound examination should be performed on women who have undergone IVF-ET, and the possibility of abdominal pregnancy should be kept in mind in the differential diagnosis of acute abdomen. PMID- 14562647 TI - Rehabilitation of occlusal support by removable partial dentures with free-end saddles. AB - This study aimed to assess the level of restored occlusal support by removable partial dentures with free-end saddles. The maximal clenching force of 61 subjects was measured using pressure sensitive film with and without their dentures. The restored level of occlusal support was estimated from the shift of the occlusal load centre. In comparison with previous results of experimental occlusions using splints for eight normal dentate subjects, the restored level using dentures ranged from 40-60% of that of natural teeth. The analysis of the shift of the occlusal load centre could reasonably evaluate the prostheses from the standpoint of occlusal support. PMID- 14562646 TI - Case report: management of tooth tissue loss from intrinsic acid erosion. AB - Acid erosive tooth wear is increasingly being reported in many western countries, and is also being recognised as a significant cause of tooth destruction in persons with xerostomia. The primary aetiology is related to a high consumption of dietary sources of acids and also to an increasing awareness of gastric acid as a significant factor in both children and adults. Recommended preventive dental treatments aim to neutralise the effects of acids, reduce the severity of xerostomia, stimulate salivary flow and buffering capacity, and increase the acid resistance of tooth substance. Initial restorative treatments should be conservative, using adhesive dentistry techniques. This treatment approach is illustrated by the oral rehabilitation of a severe erosion case using adhesive onlays, veneers and all ceramic crowns. PMID- 14562648 TI - Bonding of silicone extra-oral elastomers to acrylic resin: the effect of primer composition. AB - Silicone elastomer is bonded to acrylic resin in many facial or oro-facial prostheses. The silicone elastomer/acrylic resin bond has been reported to be insufficient and primers have been used to enhance the bond. This study investigated the bond strength of silicone elastomer to acrylic resin using different types of primers. The extra-oral silicone elastomers studied were Cosmesil and Ideal. The "overlap-joint" model was used to evaluate the bond strength and the samples were stretched until fracture. The bonding surfaces were treated with a primer. The control primer was Cosmesil and the others a mixture of Cosmesil/Z-6020 and Cosmesil/A-174 in 50/50 v/v ratio. The bond strength ranged from 0.026 MPa to 0.219 MPa. The results obtained in this work led to the conclusion that the most critical parameter allowing the efficient performance of a primer is the compatibility and affinity of its composition with the selected silicone elastomer. PMID- 14562649 TI - Comparison of bond strengths between adhesive and conventional acrylic resins to cobalt chromium denture base alloy. AB - In removable partial dentures there is normally no chemical bonding between the cast alloy and the poly(methylemethacrylate). Mechanical retention of the resin to the casting is required and presents a significant challenge during the design of partial dentures. Recently developed adhesive denture base resins which contain the monomer 4-methacryloxyethyl trimellitate anhydride can create a chemical bond to partial denture alloys. This study investigated the bond strength of adhesive denture base resin (Metafast) and conventional acrylic resin (Croform x 10) to partial denture alloy (CoCr) under three different storage conditions (dry, water, water with thermocycling). The results showed that the bond strength of the conventional acrylic resin (Croform x 10) was less than the bond strength of the adhesive denture base resin (Metafast) and the storage environment had a significant effect upon the bond strength of both resins. PMID- 14562650 TI - Satisfaction with dento-facial appearance in the eating disorders. AB - This study aimed to determine if satisfaction with dento-facial appearance is related to body image distortion and estimation in subjects with an eating disorder. Body Image Distortion, both perceived and desired, was measured with the Anamorphic Micro body image assessment software. The Body Satisfaction Scale and the Eastman Esthetic Index were used to determine satisfaction or otherwise with appearance. A convenience sample of fifteen subjects participated of whom nine were diagnosed with bulimia nervosa, two with binge eating disorder and four with eating disorder not otherwise specified. The group overestimated their body image by a mean of 28% and desired to be a mean 25% thinner. Of the 16 anatomical areas assessed on the Body Satisfaction Scale, the face was the only area associated with the over-estimate of body image (Spearman's rho = +0.52, p < 0.05). The teeth were not associated with body image. PMID- 14562651 TI - Dimensional changes and water sorption of two acrylic polymer materials reinforced with glass fibres. AB - The purpose of this study was to examine the effect of glass fibre reinforcement on the dimensional changes and water sorption of a heat-polymerising denture acrylic polymer and a microwave-polymerising denture acrylic polymer. Fifteen specimens from each group were prepared with 2% by weight (18 mg) E-glass fibre and 15 without glass fibre. The dimensional change measurements were made at three different stages on the 60 specimens. The water sorption calculations were made on specimens immersed in distilled water at 37 degrees C +/- 1 degree C for 0, 1 and 7 days. Glass fibre reinforcement and processing method do not affect the dimensional change of test specimens, but as the time increased the dimensional change decreased. The glass fibre reinforcement decreased water sorption, but the two processing methods failed to show a significant difference. PMID- 14562652 TI - Team training at an outreach dental unit. AB - A cohort of eighty nine undergraduate dental students worked for one year on two clinics completing a total of 3901 days work. Most of this time (63%) was spent at a site with shared nursing support, but despite this another site with dedicated nursing support produced approximately the same output. Students and nurses preferred working as a team rather than sharing; 98% of the students completed a questionnaire on the role of dental nurses and the majority identified chairside assistance as being the most important contribution to the increased throughput of patients. PMID- 14562653 TI - Is the NHS doing enough to promote racial equality? PMID- 14562654 TI - Should aborted foetus cells aid infertile adults? PMID- 14562655 TI - Contemporary thinking in stroke prevention and management. AB - Stroke is the third largest cause of death in the UK, after heart disease and cancer, and the largest single cause of severe adult disability (Stroke Association, 2003). Specialist and consultant stroke nurses are playing a growing role in rapid assessment clinics and acute care, as well as in coordinating rehabilitation and other follow-up services, including secondary prevention clinics. Health promotion to reduce the risk of stroke is also an important consideration and an area where nurses can play a leading role. Risk prevention measures and behaviours include physical activity, and cutting down on smoking and alcohol intake. A healthy diet is also a major factor in reducing stroke risk. This year Stoke Awareness Week, which starts on 29 September, is focusing on the importance of eating a colourful variety of fruit and vegetables, with the message: 'Eat a rainbow. Beat a stroke'. PMID- 14562657 TI - What you need to know about ... influenza. PMID- 14562656 TI - The effectiveness of surgical face masks: what the literature shows. AB - The use and withdrawal of surgical face masks in recent years has occurred in an ad hoc manner that is incompatible with evidence-based practice. Much of the literature on masks consists of anecdotal evidence or summaries of previous studies. The rationale for wearing masks has shifted from protection of the patient to protection of the health care professional wearing the mask. Currently there is little evidence that wearing a surgical mask provides sufficient protection from all the hazards likely to be encountered in an acute health care setting: the use of a respirator and face shield should be considered depending on the circumstances. PMID- 14562658 TI - A bereavement care service to address multicultural user needs. AB - A bereavement care service was set up in a large, acute NHS trust to understand and address the multicultural and multifaith needs of all its users. The service has not only facilitated support and guidance for friends and relatives at a time of great need and vulnerability, it has also made professionals throughout the trust aware of the importance of death with dignity. PMID- 14562659 TI - Pseudomyxoma peritonei. AB - Pseudomyxoma peritonei is a rare, slowly progressive disease that produces extensive mucus accumulation within the abdomen and pelvis. It is managed by cytoreductive surgery involving hyperthermic intraoperative intraperitoneal chemotherapy. PMID- 14562660 TI - Genetics. What you need to know. PMID- 14562661 TI - 'I feel honoured to be part of this'. PMID- 14562662 TI - A positive approach to Parkinson's. PMID- 14562663 TI - Developing and implementing a pain management benchmark. AB - For most patients, pain is an inevitable part of the cancer journey. Pain may be acute or chronic, it may be related to the disease process or to treatments such as surgery, radiotherapy or chemotherapy. For 80-90 per cent of patients with cancer, effective pain relief could be attained using the analgesic guidelines outlined by the World Health Organization (1996). However, in routine practice this is not achieved (Portenoy and Lesage, 1999), which highlights the need to review and improve practice continuously. PMID- 14562664 TI - Improving supportive and palliative care for adults with cancer. AB - Although many patients with cancer report positively on their experience of care, others claim they are not receiving the information and support they need at different stages of their care pathway. The national cancer patient survey (Department of Health, 2002), for example, showed wide variations in the quality of care delivered across the country. PMID- 14562665 TI - Benefits of using a chemotherapy symptom assessment scale. AB - 'Medical Intervention aims to eliminate disease, mitigate disease effect, and maximise quality of life' (Ingham and Portenoy, 1998). Diseases present with signs and symptoms: signs can be measured objectively, symptoms cannot. It is not just disease processes that cause symptoms but also the treatment of the disease. PMID- 14562666 TI - Negotiating the information maze. AB - The internet is the world's largest information source. And it is still growing rapidly--by 20 million new pages a month according to some estimates. Internet usage is also increasing. It was estimated that 140 million people were using this resource in 1996. Last year, that number had grown to an estimated 605 million (NUA Internet Surveys, 2003). One of its functions is to be a source of information for patients wanting to know more about their illnesses. PMID- 14562667 TI - Oncology is where caring really counts. PMID- 14562668 TI - Current perspectives in therapeutic myocardial angiogenesis. AB - The complex mechanisms mediating the development of new blood vessels are now beginning to be unraveled. In conjunction with major biotechnology advances, this has facilitated the initiation of translational research related to a novel treatment strategy for patients with myocardial or leg ischemia due to obstructive arterial disease--therapeutic angiogenesis. At present, at least 17 clinical trials of myocardial angiogenesis have been presented involving over 900 patients. Uncertainty exists as to the optimal delivery route and angiogenic agent, and this uncertainty is reflected in the diverse methodology of the trials published thus far. The majority of patients received an angiogenic protein via the intracoronary route. Other delivery techniques--such as direct intramyocardial injection via transepicardial or transendocardial routes--and other angiogenic agents, including master genes, have also been studied. Most recently, interest has grown in the potential angiogenesis effects of cell therapy--such as autologous bone marrow cells or cultured stem cells--and there are now several groups initiating Phase I/II trials in this area. This review summarizes the current evidence pertaining to the safety, feasibility, and efficacy of various angiogenic techniques aimed at enhancing myocardial blood flow and alleviating angina. PMID- 14562669 TI - Mortality benefit of beta blockade in patients with acute coronary syndromes undergoing coronary intervention: pooled results from the Epic, Epilog, Epistent, Capture and Rapport Trials. AB - The effects of beta blocker therapy in the settings of heart failure and coronary artery disease have been well described, although little data exist in patients presenting with acute coronary syndromes undergoing percutaneous coronary intervention. The current study will attempt to evaluate the efficacy of beta blocker therapy in this setting. Pooled data from five randomized, controlled trials of abciximab during coronary intervention were used to analyze the clinical efficacy of beta blocker therapy. The pooled analysis evaluated the end points of all-cause mortality, myocardial infarction, repeat revascularization, and the combined endpoint of death and myocardial infarction in 2,894 patients. At 30 days, death occurred in 12 of 1,939 (0.6%) patients receiving beta blocker therapy and in 19 of 955 (2.0%) patients not receiving beta blocker therapy, (P < 0.001). At 6 months, death occurred in 33 of 1,939 (1.7%) patients receiving beta blocker therapy and 35 of 955 (3.7%) not receiving beta blocker therapy, (P < 0.001). After creating a propensity model and adjusting for variables predictive of mortality in the multivariable analysis, beta blocker therapy continued to be associated with a significant reduction in mortality. The findings were similar to those shown for the effects of beta blocker therapy in separate subgroups of patients with unstable angina and acute myocardial infarction. This analysis demonstrates a lower short-term mortality in patients receiving beta blocker therapy who undergo percutaneous coronary intervention for unstable angina or acute myocardial infarction. PMID- 14562670 TI - Previous cytomegalovirus infection and restenosis after aggressive angioplasty with provisional stenting. AB - The aim of this study was to determine the impact of previous with cytomegalovirus (CMV) on restenosis after aggressive angioplasty with provisional stenting. DESIGN: We prospectively studied 78 consecutive patients scheduled for 6-month follow-up coronary angiography as part of the SIPS study. Anti-CMV IgG and IgM antibodies were measured on admission. RESULTS: Anti-CMV IgG positive and anti-CMV IgG negative patients had similar minimal lumen diameter (MLD) in the target vessel before (0.68 +/- 0.49 mm vs 0.71 +/- 0.52 mm, P = 0.84) and directly after the intervention (2.50 +/- 0.60 mm vs 2.57 +/- 0.52 mm, P = 0.58). After 6 months, however, the MLD was significantly smaller in CMV-positive as compared to CMV-negative patients (1.57 +/- 0.82 mm vs 2.00 +/- 0.83 mm, P < 0.03). Net lumen gain at 6 months was significantly lower in CMV-positive patients (0.89 +/- 0.79 mm vs 1.30 +/- 0.87 mm, P < 0.04) and the rate of clinically relevant restenosis was significantly higher (31% vs 7%, P < 0.02). In a multivariate logistic regression model, CMV seropositivity was an independent predictor of restenosis (odds ratio 5.7 (95% CI 1.2-30.3, P = 0.04). CONCLUSIONS: Six months after aggressive coronary angioplasty with provisional stenting, patients with prior CMV infection had a smaller MLD and a higher restenosis rate. CMV seropositivity was a strong independent predictor of restenosis. PMID- 14562671 TI - Acute and long-term outcomes of rotational atherectomy in small (< 3.0 mm) coronary arteries. AB - Conventional balloon angioplasty (BA) of small coronary arteries (SCA) is followed by a high rate of restenosis. Rotational atherectomy may be effective as an alternate treatment of stenoses unsuitable for other devices. The purpose of this study was to assess the efficacy of RA in the treatment of SCA. A retrospective analysis was performed of 226 lesions in 159 consecutive patients who underwent RA of SCA (mean diameter = 2.36 +/- 0.49 mm). One hundred forty eight lesions (65.5%) were type B2 or C of AHA/ACC criteria. Follow-up angiography was performed at 3 and 6 months after the procedure. Procedural success was achieved in 96.9% of patients. The mean burr-to-artery ratio was 0.74 +/- 0.17. Adjunctive BA and stent implantation were needed in 94.2% and 22.6% of lesions, respectively. Minimal lumen diameter (MLD) increased from 0.66 +/- 0.35 mm to 1.97 +/- 0.58 mm (P < 0.01). Angiographic complications consisted of acute reclosure (3.5%), no reflow/slow flow (12.4%), and coronary artery perforation (1.8%). No death, Q-wave myocardial infarction (MI), or coronary artery bypass graft (CABG) occurred during the initial hospitalization. Restenosis rates at 3 and 6 months were 40.6% and 44.2%, respectively, and target lesion revascularization (TLR) rates were 28.5% and 33.0%, respectively. Restenosis and TLR rates during follow-up were comparable among patients who underwent RA + adjunctive BA versus patients who underwent RA + stenting. Long-term clinical follow-up was complete in 143 patients over a mean period of 348 +/- 166 days. The survival free from cardiac death, MI, CABG or repeated BA was 59.6% at 1 year. In conclusion, RA of SCA has relatively high restenosis rates, but may be appropriate for more complex, calcified lesions unsuitable for other devices. PMID- 14562672 TI - The artery: half full or half empty? PMID- 14562673 TI - Comparison of a silicon carbide coated stent versus a noncoated stent in humans: the Tenax- versus Nir-Stent Study (TENISS). AB - PURPOSE: Stents coated with hypothrombogenic silicon carbide (a-SiC:H) exhibited low restenosis rates in the rabbit and in an observational study in humans. Thus, the clinical and angiographic outcome was assessed in a large multicenter study. MATERIAL AND METHODS: Four hundred and ninety-seven patients (63.4 +/- 9.8 years) were randomized to either receive the a-SiC:H-coated Tenax stent or the stainless steel Nir stent. Lesions (diameter > or = 2.8 mm, length < 20 mm) were covered with one single stent. RESULTS: Fifty-one of 497 (10.3%) patients were excluded for protocol violation. Three hundred and forty-two of 446 (76.7%) patients presented for scheduled angiographic follow-up after 4.7 +/- 1.2 months and 29 of 446 (6.5%) prematurely. In-hospital complications comprised two deaths (0.8%) (P > 0.99) and one (0.4%) (P > 0.99) CK-elevation in each group, target lesion revascularization in 5 of 250 (2%) of the Tenax and 4 of 244 (1.6%) of the Nir sample (P > 0.99), and subacute thrombosis in 2 of 250 (0.8%) of the Tenax patients (P = 0.5). In the Tenax/Nir patients mean percent diameter stenosis decreased from 82.3 +/- 9.1%/80.7 +/- 8.4% (P = 0.49) to 17.6 +/- 5.5%/17.6 +/- 5.5% (P = 0.99) postprocedure and increased to 34.5 +/- 21.5%/34.2 +/- 23.1% (P = 0.90) at follow-up. CONCLUSIONS: Thus, there appears to be no advantage of the silicon carbide coated stent over a stainless steel stent after 4.7 +/- 1.2 months with regard to clinical and angiographic restenosis rates. PMID- 14562674 TI - Interventional occlusion of a large pulmonary arteriovenous malformation with an Amplatzer septal occluder. AB - Interventional occlusion of pulmonary arteriovenous malformations with large feeding arteries may be associated with an increased risk of device embolization. In a 40-year-old patient with a solitary pulmonary arteriovenous malformation one large feeding artery was successfully closed by means of an Amplatzer septal occluder and detachable coils. The use of an Amplatzer septal occluder should be considered as an alternative therapeutic option in cases of huge pulmonary arteriovenous malformation to reduce the risk of device embolization. PMID- 14562675 TI - Another arrow in the quiver--but at what cost? PMID- 14562676 TI - Percutaneous, catheter-based coil embolization of coronary fistula: determination of hemodynamic relevance. AB - A 55-year-old male with angina-like chest pain and positive thallium-scintigraphy was admitted to our hospital. Cardiac catheterization was performed for suspected coronary artery disease. The coronary angiogram showed no significant epicardial stenosis, but a large coronary fistula, connecting the left anterior descending artery with the pulmonary artery. Swan-Ganz catheter measurements, intracoronary Doppler, and quantitative coronary angiography were used to determine cardiac output, coronary blood flow, and coronary-to-pulmonary artery shunt fraction. These measurements showed a hyperdynamic cardiac output of 17 L/min, a coronary blood flow of 140 mL/min in the left anterior descending coronary artery with an estimated shunt fraction of 58% into the pulmonary circulation. Percutaneous, catheter-based coil embolization was performed to occlude the fistula. After embolization of one coil, coronary angiography showed the fistula's stump only. Cardiac output (9 L/min) and coronary blood flow (48 mL/min) were almost normalized. The patient was discharged from the hospital the day after the procedure. After a 6-month follow-up, there were still no complaints, angina-like symptoms or signs of myocardial ischemia in stress tests. PMID- 14562677 TI - Microcoil embolization for treatment of a right coronary arteriovenous fistula. AB - A patient initially presented with anginal symptoms and a positive stress thallium test. An arteriovenous malformation in the right coronary artery causing a suspected coronary "steal syndrome" was subsequently discovered. This was treated with a microcoil embolization technique in lieu of the traditional surgical approach and this technique is described in detail. The patient had successful clinical, nuclear, and angiographic outcomes. PMID- 14562678 TI - Closure of a femoral pseudoaneurysm by transcutaneous short guidewire embolization. AB - Femoral pseudoaneurysm (PSA) is a well-recognized complication following catheter based diagnostic or interventional procedures. We report a patient who developed a pseudoaneurysm following the insertion of a Steinman pin for a fractured shaft of the right femur following a road traffic accident. The treatment of choice for a femoral pseudoaneurysm is ultrasound-guided compression repair. As this modality of treatment was not applicable to our patient, the pseudoaneurysm was closed by transcutaneous short guidewire embolization technique. PMID- 14562679 TI - [Computer-assisted vaccine design]. AB - With the modern molecular biology techniques, it has been possible to detect, isolate and clone biological macromolecules, which could be used as immunogenes in artificial vaccine constructs. In the post-genomic era, the prospective immunogenic components are searched using bionformatic tools and proteomic technologies. Today it is quite realistic to combine the artificial vaccine constructs from the preselected molecular components. Existing computational methods are able to detect the potential immunogenes in genomic sequences, predict their characteristics and subcellular location. The set of methods is designed to predict the T- and B-epitopes that can be used as components of minimal vaccine constructs. The variety of systems for production and delivery of vaccines are developed and tested. These include transgenic plants, bacterial and viral vectors, DNA molecules etc. Several informational resources provide free access to molecular immunology data and deliver services on prediction of antigenic features. Several artificial vaccines have already been launched, but much more preparations are under preclinical and clinical trials. Computer-aided design of vaccines may significantly decrease time and costs required for their development. Modern bioinformatic technologies are now employed for discovery of more effective and potent vaccine. PMID- 14562680 TI - [Apoptosis modulators]. AB - At least two types of cell death are known: physiological (apoptosis) and pathological (necrosis). Dysregulation of apoptosis plays an important role in the pathogenesis of many pathological conditions. Their analysis gives a key for the development of novel therapeutic approaches. In this review the recent advances and perspectives for clinical use of apoptosis-modulating monoclonal antibodies, low-molecular-weight proteins, peptides, small non-peptide molecules and drugs targeting apoptosis-associated genes are summarized. The rationale for clinical application of apoptomodulators with the aim of increasing efficacy of existing standard therapies are also discussed. PMID- 14562681 TI - [Vascular endothelial growth factor: its secretion in the bone tissue in the norm and in pathological states]. AB - Vascular endothelial growth factor (VEGF) is a homodimeric heparin-binding protein (34-42 kDa), which induces formation of new blood vessels (angiogenesis). VEGF-mediated capillary invasion is an essential signal that regulates growth plate morphogenesis and triggers cartilage remodeling. Thus, VEGF is essential coordinator of chondrocyte death, chondroclast function, extracellular matrix remodeling, angiogenesis and bone formation in the growth plate. VEGF expression in untreated osteosarcoma is predictive for pulmonary metastasis and poor prognosis. VEGF concentration in the serum of patients with malignant bone tumors (osteosarcoma, Ewing's tumor) increased in comparison with norm and benign tumors. VEGF expression in the serum of patients with osteosarcoma associated with the development of metastasis and without metastasis period. The results of investigation indicate the existence of dependence between VEGF expression and degree of tumor malignancy and metastasis. These results testify probable participation of VEGF in biology of the tumor-transformed tissue. PMID- 14562682 TI - [Metabolism of endogenous aldehydes: participation in oxidative stress-induced lesions and its age aspects]. AB - This review summarizes the data regarding the synthesis and utilization of endogenous aldehydes and mechanisms of their cytotoxic effect. Peroxidation of polyunsaturated fatty acids is the main source of endogenous aldehydes. There are many different aldehydes generated in the cell. The most abundant is 4-hydroxy 2,3-nonenal synthesized from linoleic acid. Aldehydes may react with proteins and nucleic acids and change their functional properties. Aldehyde utilization mainly occurs in reactions catalysed by aldehyde dehydrogenase, aldehyde reductase and glutathione-S-transferase. The major pathway of their catabolism is accompanied with their conjugation with glutathione. Endogenous aldehyde utilization has its tissue- and age-dependent specificity. The status of aldehyde catabolism can modulate free radical alteration effect on the cell. An adequate stimulation of endogenous aldehyde utilization in the situation of enhancement of free radical generation may promote increasing the cell resistance to oxidative stress injury. Senescence is accompanied by a decrease in endogenous aldehyde utilization intensity in tissues. This could be important in the pathogenesis of age dependent pathology. PMID- 14562684 TI - [Effects of Gerbamarin T balms on functions of the cardiovascular and hepatobiliary systems]. AB - The effects two balms of "Herbamarin" T series on some parameters of cardiovascular and hepatobiliary systems were studied in mice either kept on the cholesterol diet or treated with CCl4. The results allow to recommend these balms as additional components to the therapy of various cardiovascular and hepatic diseases. PMID- 14562683 TI - [mRNA level and cytochrome P450 1A activity in the liver of C57BL mice induced by various xenobiotics]. AB - The rate of hepatic cytochrome P450 Cypla1 and Cyp1a2 induction was investigated in C57BL male mice during induction with o-aminoazotoluene (OAT), benzo[a]pyrene (BP) and 1,4-dihydroxyanthraquinone (AQ). The Cypla1 mPNA level determined by quantitative RT-competitive PCR increased more than three orders of magnitude during induction with OAT and BP compared with untreated animals and remained unchanged during induction with AQ. The Cypla2 mRNA level was only 8.5, 18.7 and 1.9 times higher during induction with OAT, BP and AQ respectively than in untreated mice. At the same time 7-Ethoxyresorufin-O-deethylase (EROD) and 7 Methoxyresorufin-O-demethylase (MROD) activities of Cypla were also investigated in liver. The increase of Cypla1 mRNA level correlated with the increase of EROD activity. This suggests involvement of the transcriptional mechanism of the inducibility of this enzyme. In the case of Cypla2 there was insignificant increase of its mRNA level but high catalytic activity registered in liver in response to injection of the inductor of MR metabolism. This can imply the posttranscriptional mechanism of Cypla2 regulation. During induction with AQ the Cypla1 mRNA level remained unchanged, but the EROD activity increased almost 20 fold. The latter suggests insufficient specificity of this substrate for Cypla1. Thus, on the basis of the data obtained, the mRNA level can be considered as more accurate estimation of Cypla1 and Cypla2 inducibility, than the determination of the enzyme activity. PMID- 14562687 TI - [Dog-days and the elderly. The killing summer]. PMID- 14562685 TI - [Blood group-specific variability of sialic acid contents in the chorionic gonadotrophin structure]. AB - Low-pressure ion-exchange chromatography was used for isolation of chorionic gonadotropin (hCG) from urine of pregnant women. The purified hCG-preparations from various blood group donors were shown to differ in their sialic acid content. The highly purified hCG-preparations after gel chromatography were analyzed in the reaction of hemagglutination and the preliminary results suggest their blood group specificity. PMID- 14562688 TI - [The elderly, from compassion to action]. PMID- 14562689 TI - [Home services and natural disasters]. PMID- 14562690 TI - [Which public policy for elderly handicapped persons?]. PMID- 14562691 TI - [History and demography of elderly handicapped persons]. PMID- 14562692 TI - [Aging of handicapped persons, the challenge of a new gerontology]. PMID- 14562693 TI - [Rhone Information and Social Action Center, an association in the heart of departmental dynamics]. PMID- 14562694 TI - [Reconstituting an enlarged family, the innovative Hameau Service project]. PMID- 14562695 TI - [With the advance of age, what solutions for the mental deficient person?]. PMID- 14562696 TI - [Facing Alzheimer's disease. 6/6--support for families]. PMID- 14562697 TI - [Drug distribution in nursing homes, a green light for nurses' aides]. PMID- 14562698 TI - [How to use pain measurement instruments?]. PMID- 14562699 TI - Measurements of oxygen in tissues: overview and perspectives on methods. AB - The goal of this manuscript is to provide a summary of the major techniques that currently are being applied to measure oxygenation of tissues in vivo. Oxygen is one of the key components of metabolism. Oxygen is also a major variable in many diseases, both with respect to the pathophysiological processes and influencing the efficacy of treatment. Unfortunately, however, the measurement of tissue oxygenation is non-trivial. Consequently many different methods have been developed to try to make this measurement. This paper presents a summary, largely in tabular form, of most of the current methods for assessing tissue oxygenation. The table attempts to cover the most pertinent aspects of the techniques and their applications, including their potential niche, limitations, and advantages. Citations are given for each method to point the reader in the direction of relevant literature. PMID- 14562700 TI - Quantitative brain tissue oximetry, phase spectroscopy and imaging the range of homeostasis in piglet brain. AB - The quantification of tissue oxygen by frequency or time domain methods has been discussed in a number of prior publications where the meaning of the tissue hemoglobin oxygen saturation was unclear and where the CW instruments were unsuitable for proper quantitative measurements [1, 2]. The development of the IQ Phase Meter has greatly simplified and made reliable the difficult determination of precise phase and amplitude signals from brain. This contribution reports on the calibration of the instrument in model systems and the use of the instrument to measure tissue saturation (StO2) in a small animal model. In addition, a global interpretation of the meaning of tissue oxygen has been formulated based on the idea that autoregulation will maintain tissue oxygen at a fixed value over a range of arterial and venous oxygen values over the range of autoregulation. Beyond that range, the tissue oxygen is still correctly measured but, as expected, approaches the arterial saturation at low metabolic rates and the venous saturation at high metabolic rates of mitochondria. PMID- 14562701 TI - Tumor oximetry: comparison of 19F MR EPI and electrodes. AB - We recently described a novel approach to measuring regional tumor oxygen tension. This approach is based on 19F pulse burst saturation recovery NMR echo planar imaging relaxometry of hexafluorobenzene or "FREDOM" (Fluorocarbon Relaxometry using Echo planar imaging for Dynamic Oxygen Mapping). We have now compared oxygen tension measurements using FREDOM with a traditional polarographic method (the Eppendorf Histograph) in a group of size matched Dunning prostate rat tumors R3327-AT1. We also compare MR and electrode approaches to monitoring dynamic changes with respect to interventions and demonstrate extension of the MR technique to rat breast tumors. PMID- 14562702 TI - Mapping cerebral glutamate 13C turnover and oxygen consumption by in vivo NMR. AB - Regional rates of 13C incorporation from glucose to glutamate were detected in anesthetized rat brain in vivo at 7T with high temporal and spatial resolution using NMR method ICED PEPSI (in vivo carbon edited detection with proton echo planar spectroscopic imaging). Time courses of regional glutamate 13C turnover were fitted by a metabolic model to obtain regional tri-carboxylic acid (TCA) cycle flux and cerebral metabolic rate of oxygen consumption (CMRO2) in each voxel (8 microL) of rat cortex. CMRO2 maps obtained for rats under either alpha chloralose or morphine anesthesia revealed average cortical values of 1.5 +/- 0.2 (n = 3) and 3.2 +/- 0.3 (n = 4) mumol/g/min, respectively. These values of CMRO2 are in good agreement with previous cortical measurements with coarser spatial resolution. The heterogeneity within each map, which depicted predominantly gray and white matter differences, was significantly greater under morphine (higher cortical activity) than under-alpha-chloralose (lower cortical activity) anesthesia. The regional variations in the basal awake state, which are expected to be even greater, should be considered to avoid partial-volume artifacts in functional activation studies of awake subjects. PMID- 14562703 TI - Monte-Carlo simulation of light transport for NIRS measurements in tumors of elliptic geometry. AB - Propagation of light in a highly scattering medium such as biological tissue is difficult to study. For complex geometry and multilayer structures computer simulation has to be used for light transport analysis. A Monte Carlo model of light propagation in tissue has been applied for the purpose of better understanding of the results of near-infrared spectroscopy (NIRS) measurements in experimental tumors. The major objective was to determine the percentage and location of the illuminated area in tumor and to estimate fraction of NIRS signal originating from the underlying tissues. Values of optical parameters used in the model were taken from literature. Tumor shape was approximated with a rotational ellipsoid. Computer simulations were made for two positions of optodes: reflectance and transmittance mode. Results of simulations indicate that in both configurations the majority of signal originates from tumor and not from surrounding tissue. In reflectance mode collected light comes from limited area near the optode whereas in transmittance mode the collected light illuminate almost whole tumor. This difference between the two modes is valid for all tissue parameters. PMID- 14562704 TI - Arterial pulsations are present in one third of the human cranial vascular volume penetrated by near infrared light. AB - The percentage of cranial vascular volume that undergoes arterial pulsations is estimated by 18 paired measurements in nine healthy volunteers using near infrared spectroscopy (NIRS, 10 Hz sampling frequency, 769 nm). The NIR absorption is decomposed in pulsatile and non-pulsatile components by digital filtering. The time course of absorption changes of these components after sudden intravenous injection of indocyanine-green (ICG) is used to estimate the arterial fraction of vascular volume (f(art)). Approximately 28% of the vascular volume within the optical field of the NIR spectroscope was arterial. The range of values was 6-43%. Variance of f(art) was 4.8 times higher between subjects than within subjects, indicating that the variability observed is not due to imprecise estimation but to optode position and subject. PMID- 14562705 TI - Absolute frequency-domain pulse oximetry of the brain: methodology and measurements. AB - A new method to non-invasively measure the absolute tissue oxygen saturation (SO2) and arterial oxygen saturation (fdSaO2) by frequency-domain spectroscopy is described. This method is based on the quantitative measurement of the tissue absorption spectrum, which is used to determine global SO2. From the amplitude of absorption changes caused by arterial pulsation oscillations, in the range of 633 841 nm, the fdSaO2 can be calculated. During deoxygenation (air/N2 mixture) experiments, we measured the fdSaO2 and SO2 on the forehead of three healthy volunteers and compared them to the arterial oxygen saturation measured by conventional pulse oximetry (poSaO2) on the finger. fdSaO2 and poSaO2 agree very well (mean difference: -1.2 +/- 2.6%). Changes in SO2 were systematically smaller than in fdSaO2 or poSaO2 probably due to autoregulation. The measurements with 4 and 8 wavelengths had comparable quality. PMID- 14562706 TI - The influence of a clear layer on near infrared spectrophotometry: comparison of measurements in a liquid neonatal head phantom to infants in vivo. AB - Near infrared spectrophotometric (NIRS) algorithms to determine the tissue oxygen saturation (TOI) assume a semi-infinite, homogenous tissue geometry. At the head, the clear cerebrospinal fluid (CSF) layer may violate this assumption. The aim was to estimate the error in the TOI values caused by the CSF layer in vitro and to confirm the results in vivo. The liquid phantom mimicking the neonatal head, consisted of a spherical shell of silicone filled with a liquid solution (1% Intralipid, 60 mumol/l haemoglobin, yeast) and a clear layer imitating CSF. The solution was oxygenated and deoxygenated, while measuring its TOI and pO2. Without clear layer the mean TOI was 90.9 +/- 0.5% at pO2 > 18 kPa and decreased to 26.0 +/- 1.3% at pO2 = 0 kPa. With a clear layer the TOI increased from 27.8 +/- 0.8% at pO2 > 18 kPa to 68.0 +/- 0.8% at pO2 = 0 kPa. The clear layer caused a large error in the TOI. In ten mechanically ventilated infants (postnatal age 0.03 to 8 months) the TOI (at the head) and arterial oxygen saturation (SaO2) were measured while the inspired oxygen fraction was altered. The TOI was always positively correlated with the SaO2 (mean slope linear regression = 0.89, r2 = 0.62). Thus an adverse effect of the CSF layer on TOI measurements can be excluded for infants. The CSF layer is not modelled correctly in the phantom. PMID- 14562707 TI - Strategies for absolute calibration of near infrared tomographic tissue imaging. AB - Quantitative near infrared (NIR) imaging of tissue requires the use of a diffusion model-based reconstruction algorithm, which solves for the absorption and scattering coefficients of a tissue volume by matching transmission measurements of light to the predictive diffusion equation solution. Calibration problems as well as other practical considerations arise for an imaging system when using a model-based method for a real system. For example, systematic noise in the data acquisition hardware and source/detector fibers must be removed to prevent spurious results in the reconstructed image. Practical considerations for a NIR diffuse tomographic imaging system include: (1) calibration with a homogeneous phantom, (2) use of a homogenous fitting algorithm to arrive at an initial optical property estimate for image reconstruction of a heterogeneous medium, and (3) correction for fluctuations in source strength and initial phase offset during data acquisition. These practical considerations, which rely on an accurate homogeneous fitting algorithm are described. They have allowed demonstration of a prototype imaging system that has the ability to quantitatively reconstruct heterogeneous images of hemoglobin concentrations within a highly scattering medium with no a priori information. PMID- 14562708 TI - Brain tissue and sagittal sinus pO2 measurements using the lifetimes of oxygen quenched luminescence of a ruthenium compound. AB - The study was done to assess the performance of a system that measures the partial pressures of oxygen (pO2) from the lifetimes of oxygen-quenched luminescence of ruthenium compounds immobilized at the tip of fiber-optic optodes (Oxylite system). The system was used to measure the pO2 in brain tissue (thalamus and hypothalamus) and in the sagittal sinus of isoflurane-anesthetized rats at different FiO2's. The pO2 recorded in the hypothalamus (HPtO2) was consistently higher than the pO2 in the thalamus (TPtO2) at all FiO2. HPtO2 was closely related to PvO2 during normoxia but not during hypoxia. The equilibrium time of Oxylite system was found to be rapid compared to in vivo tissue response to changes in FiO2. PMID- 14562709 TI - Preliminary studies on the photon path in breast tissue model by NIR-TRS. AB - A typical tumor releases angiogenesis factor that induces the capillary growth around the tumor and, therefore, a greater amount of blood is present around the tumor. A tumor usually, therefore, becomes a local absorber due to its higher hemoglobin concentration. The ultimate goal of this project is to localize the position of a tumor in a breast tissue at an early stage using near infrared spectroscopy. Computer simulations were performed to obtain TRS spectra at various locations in a system with optical properties of human breast. The time domain output pulses, then, were transferred to the frequency domain and the data were analyzed at various modulated frequencies. The changes in the photon path with respect to the frequency were systematically studied for absorber localization in three-dimensions. Two different source-detector configurations, transmittance and reflectance were studied to explore the best TRS spectra acquisition procedure for tumor localization. Our previous study results have shown that in reflectance measurements the photon penetration depth is dependent on both the source-detector (S-D) separation distance and the modulation frequency. More specifically, the photon penetration depth decreases at smaller source and detector separations and higher frequencies. In this study, the effects of these two parameters on the photon path were studied in both transmittance and reflectance. This study results may be used effectively in determining the position. PMID- 14562710 TI - EPR spectroscopy and imaging of oxygen: applications to the gastrointestinal tract. AB - EPR imaging has emerged as an important tool for noninvasive three-dimensional (3D) spatial mapping of free radicals in biological tissues. Spectral-spatial EPR imaging enables mapping of the spectral information at each spatial position, and, from the observed linewidth, the localized tissue oxygenation can be mapped. We report the application of EPR imaging techniques enabling 3D spatial and spectral-spatial EPR imaging of small animals. This instrumentation, along with the use of a biocompatible charcoal oximetry-probe suspension, enabled 3D spatial imaging of the gastrointestinal (GI) tract, along with mapping of oxygenation in living mice. By using this technique, the oxygen tension was mapped at different levels of the GI tract from the stomach to the rectum. The results clearly show the presence of a marked oxygen gradient from the proximal to the distal GI tract, which decreases after respiratory arrest. This technique for in vivo mapping of oxygenation is a promising method, enabling the noninvasive imaging of oxygen within the normal GI tract. This method should be useful in determining the alterations in oxygenation associated with disease. PMID- 14562711 TI - Effects of blocking buffers and plasma proteins on the protein C biosensor performance. AB - Protein C (PC) deficiency can lead to abnormal thrombus formation in blood vessels, obstructing oxygen and nutrient transport to various organs or tissues. Quantifying PC amount in blood plasma usually takes long time, is difficult and expensive due to its low concentration and other homologous proteins in it. A fiber-optic immunosensor has been under development for several years for the PC real-time assay. In this study, blocking buffers have been examined to reduce the noise caused by non-specific adsorption. Ethanolamine was found to reduce the noise significantly. The effect of various PC homologues (factors II, VII, IX, and X) on the biosensor performance was investigated. The sensor showed little cross reactivity with these homologues. Human serum albumin (HSA) in the sample decreased the signal intensity. However, PC could be still quantified in samples with HSA at the physiological level. PMID- 14562712 TI - Protein C separation from human blood plasma derivatives using low cost chromatography. AB - Protein C (PC) deficiency can cause thrombosis, inhibiting oxygen transport to tissue thus resulting in many complications, including death. Present treatment can cause catastrophic bleeding and other major medical problems. PC treatment has no bleeding or skin necrosis problems because it circulates in the blood as a zymogen and is only activated when and where it is needed. The vitamin K dependent (VKD) proteins are homologous proteins, making the separation of PC from plasma extremely difficult. Immobilized metal affinity chromatography (IMAC) is investigated to separate the VKD proteins to replace immunoaffinity chromatography, because of the high cost of monoclonal antibodies. An IDA-Cu column was found effective for the separation of PC from prothrombin, the most harmful contaminant. For Cohn fraction IV-1 separation, a DEAE column was found an efficient initial step, with about 25-fold PC purity increase. Following this step, an IDA-Cu column could remove many contaminants including prothrombin. The combination of DEAE and IDA-Cu resulted in PC purity increase of about 100-fold. PMID- 14562713 TI - Glutathione depletion or radiation treatment alters respiration and induces apoptosis in R3230Ac mammary carcinoma. AB - Glutathione depletion by L-buthionine sulfoximine inhibits the growth of Ehrlich mouse mammary carcinoma, R3230Ac rat mammary carcinoma and the PC3 human prostrate carcinoma cells, in vitro. Inhibition of growth occurs within the first 24 hours after exposure to the drug. The cell density does not increase over the initial cell density over 7 days. A549 human lung carcinoma and the DU145 human prostrate carcinoma cells show no inhibition of growth under the same treatment conditions. A comparative study of the R323OAc and A549 cells demonstrated a marked increase in apoptosis following L-BSO treatment in R3230Ac, which was dependent on L-BSO concentration and incubation time. L-BSO did not induce apoptosis in A549 cells at any of the concentrations tested. The incidence of apoptosis for R323OAc cells following exposure to 0.1 mM L-BSO was similar to the incidence of radiation-induced apoptosis observed after exposure to 10 Gy. Treatment with L-BSO or radiation alone inhibited O2 utilization in of R323Oac, while no effect on O2 utilization was observed in A549 cells. LBSO altered the bioreductive capacity of both the R323OAc and A549 cells. These results suggest that the ability of L-BSO to block mitochondrial O2 utilization may be involved in the apoptotic response in R3230Ac cells. PMID- 14562714 TI - Oxygenation and vascular perfusion in spontaneous and transplanted tumor models. AB - Since quantitative measurements of tumor vascular function cannot be obtained in human tumors, appropriate animal tumor models must be utilized. The current studies were undertaken to compare transplantable, murine KHT tumors with primary and 1st generation transplants of spontaneous mammary carcinomas. To evaluate changes in tumor vascular structure and function, immunostaining of total and perfused vascular spacing, and cryospectrophotometric measurement of intravascular HbO2 saturations were utilized. KHT tumors demonstrated a distinct pattern of decreasing oxygenation with increasing distance from the tumor surface, while spontaneous tumors exhibited striking intertumor heterogeneities and a reduced dependence of oxygenation on distance from tumor surface. Anatomical/perfused vessel distributions and functional response were similar between the primary and transplanted tumor models, as was tissue histological appearance, but were quite different from KHT tumors. These results indicate that spontaneous tumor vascular configuration and function tend to be preserved in 1st generation trochar transplanted tumors. PMID- 14562715 TI - Effect of mild hyperglycemia +/- meta-iodo-benzylguanidine on the radiation response of R3230 Ac tumors. AB - The effects of glucose or meta-iodo-benzylguanidine (MIBG) on oxygen utilization (QO2) of several tumor cell lines were studied using a Clark-type electrode chamber. For in vivo studies, rats bearing R3230 Ac rat mammary adenocarcinomas were utilized. To evaluate changes in tumor oxygenation induced by glucose or MIBG, intratumoral pO2 and skeletal muscle pO2 were measured using Eppendorf Histography. To find the effect of mild hyperglycemia (i.p., 1 g/kg) +/- MIBG (i.p., 20 mg/kg) on the radiation response, a growth delay assay was used. Glucose alone produced a approximately 20% inhibition of QO2 in several tumor cells we tested except Q7 tumor cells. MIBG inhibited QO2 in R3230 Ac tumors. The median tumor pO2 for glucose + MIBG was increased from 5.3 mm Hg to 13.8 mm Hg. We hypothesized that combined treatment with glucose + MIBG significantly enhanced radiation-induced tumoricidal effects on R3230 Ac tumors, mainly due to reduction in QO2 and increase in tumor pO2. PMID- 14562716 TI - Effect of ONCONASE +/- tamoxifen on ASPC-1 human pancreatic tumors in nude mice. AB - To evaluate changes in tumor physiological parameters after treatment with ONCONASE (ONC), we used laser Doppler flowmetery for tumor blood flow (TBF) in legs of nude mice bearing AsPC-1 human pancreatic carcinoma. Tumor interstitial fluid pressure (TIFP) was measured using the wick-in-needle technique, while intratumoral pO2 utilized O2 sensitive needle electrodes. TBF was significantly increased by an i.p. injection of ONC, then returned to the untreated levels at 150 min post-treatment. Single and multiple intraperitoneal injections of ONC significantly reduced TIFP (post-treatment at 1-7 days). ONC significantly improved tumor oxygenation evidenced by an increase in median pO2 from 4.2 mm Hg to 8.2 mm Hg. Since ONC had a synergistic interaction with tamoxifen (TAMX) on the cytotoxic clonogenicity of AsPC-1 tumor cells in vitro, we evaluated the antitumoral effects in vivo by ONC +/- TAMX, using nude mice bearing AsPC-1 pancreatic tumor cells of ascite (intraperitoneally implanted) or solid (subcutaneously implanted to legs) tumors. ONC alone effectively retarded the tumor growth, but TAMX alone did not. ONC + TAMX was synergistically effective in inhibiting tumor growth. PMID- 14562717 TI - Oxygenation in a human tumor xenograft: manipulation through respiratory challenge and antibody-directed infarction. AB - We recently demonstrated the use of 19F NMR relaxometry of hexafluorobenzene to monitor regional tumor oxygen tension dynamics in rats. We have now extended the application to human tumors implanted in immunocompromised (SCID) mice. This has allowed us both to investigate dynamic response to respiratory challenge (carbogen) and to probe the mechanisms of a new anti-vascular therapy designed to produce tumor-specific infarction. PMID- 14562718 TI - Tumor pO2 assessments in human xenograft tumors measured by EPR oximetry: location of paramagnetic materials. AB - Radioantibody immunotherapy (RAIT) is a promising treatment modality but the effectiveness of this targeted low dose radiation varies from tumor to tumor. Since RAIT is an oxygen dependent treatment, baseline pO2 or growth-induced changes in the microenvironment may alter treatment response. In this pilot work we monitored tumor pO2 in untreated human xenograft tumors growing s.c. in nude mice. These data will be used to plan a study of the relationship between the effectiveness of RAIT and tumor pO2. Growth or treatment-induced changes in the microenvironment may alter the tumor pO2 and thus affect the response to therapy but may also affect location and microenvironment of the particulate oxygen sensor. We monitored tumor pO2 during growth and also examined the tumor histological structure overall and in the region of the paramagnetic material in the tumor at the time of necropsy. PMID- 14562719 TI - Hemoglobin imaging with hybrid magnetic resonance and near-infrared diffuse tomography. AB - This study examines the methodology of combining high-resolution information from magnetic resonance imaging into the reconstruction of near-infrared images of hemoglobin concentration and oxygen saturation. This type of hybrid imaging modality has the potential to provide noninvasive maps of hemoglobin concentration and oxygen saturation with relatively high spatial resolution with a fast time response. The study uses (i) tissue-simulating phantoms, as well as (ii) a rat cranial model, to test the method in two well-controlled situations. The phantom test demonstrates that better reconstruction accuracy can be achieved with the use of MRI-generated spatial regions in near-infrared reconstruction. The rat functional testing reveals that the technique can be applied to in vivo physiology, even in situations where the tissue is quite heterogenous. It also shows that the application of a priori structure in the finite element mesh as well as spatial constraints in the near-infrared image reconstruction, can significantly improve the quality of the resulting hemoglobin images. PMID- 14562721 TI - Microcirculatory function, tissue oxygenation, microregional redox status and ATP distribution in tumors upon localized infrared-A-hyperthermia at 42 degrees C. AB - Since local hyperthermia (HT) affects microenvironmental parameters, the aim of the study was to analyze the impact of 42 degrees C-HT on microcirculatory function, tumor pO2, microregional redox status and ATP distribution in experimental rat tumors. Subcutaneously growing DS-sarcomas were treated with localized HT using infrared-A radiation resulting in a mean tumor temperature of 42 degrees C. The relative red blood cell (RBC) flux in the tumor was assessed using the laser Doppler technique and the mean tumor pO2 measured continuously using O2-sensitive catheter electrodes. In a second series of experiments, the microregional distribution of the mitochondrial redox status and ATP concentration was measured. Although the average RBC flux increased by 63%, tumor pO2 rose only by approx. 6%. No distinct changes were seen in the mitochondrial redox status. The microregional distribution of the redox status as well as of the ATP concentration showed considerable heterogeneity. In conclusion, although 42 degrees C-HT leads to a distinct improvement in tumor perfusion, there is practically no change in the oxygenation status. The latter finding can be explained by an equivalent increase in the oxygen consumption rate of the cells which increases by approx. 58% at 42 degrees C compared to normothermia. PMID- 14562720 TI - Tumor oxygen dynamics: comparison of 19F MR EPI and frequency domain NIR spectroscopy. AB - Oxygen plays a key role in tumor therapy and may be related to tumor development: e.g., angiogenesis and metastasis. Using noninvasive techniques to accurately measure tumor oxygenation could assist in developing novel therapies. Here, we have used the FREDOM (Fluorocarbon Relaxometry using Echo planar imaging for Dynamic Oxygen Mapping) approach based on hexafluorobenzene (HFB) to monitor tissue oxygen tension (pO2) of rat breast and prostate tumors and compared the results with changes in tumor vascular hemoglobin saturation (sO2) and concentration observed using a new dual wavelength homodyne near-infrared (NIR) system. The dynamic changes in pO2 and sO2 were assessed while rats were breathing various gases. NIR showed significant changes in vascular oxygenation accompanying respiratory interventions. 19F MR-EPI also showed significant changes in tissue pO2 and revealed considerable regional heterogeneity in both absolute values and rate of change accompanying interventions. Generally, changes in vascular sO2 preceded tissue pO2, particularly for smaller tumors. PMID- 14562722 TI - Allosteric modification of hemoglobin by RSR13 as a therapeutic strategy. AB - RSR13 binds to hemoglobin (Hb), reduces oxygen (O2) binding affinity, and enhances O2 unloading from Hb to hypoxic tissue. Tissue hypoxia is common to cancer, surgery, myocardial ischemia, and stroke. RSR13 increases tumor pO2, reduces tumor hypoxic fraction and because O2 is necessary to maximize the effectiveness of radiation therapy, RSR13 enhances the efficacy of radiation therapy. Patients with brain metastases or glioblastoma multiforme receiving RSR13 and radiation therapy have improved median survival, compared to matched historical controls. Myocardial and cerebral hypoxia can be complications to cardiopulmonary bypass (CPB) surgery. RSR13 improves myocardial oxidative metabolism and contractile function in models of myocardial ischemia, including CPB. In CPB patients, RSR13 improved cardiac contractile function and reduced blood product use. In animals, RSR13 increased brain pO2 and reduced neuronal cell death following cerebral ischemia, alone or in combination with excitotoxic neurotransmitter inhibition. Allosteric modification of Hb by RSR13 represents a unique therapeutic strategy. PMID- 14562723 TI - The pharmacology of tissue oxygenation by biopure's hemoglobin-based oxygen carrier, Hemopure (HBOC-201). AB - Biopure's hemoglobin-based oxygen carrier, HBOC-201 (Hemopure), enhances oxygen transport by promoting both the convective and diffusive components of transport in the microcirculation. Convective transport is modified by HBOC-201 in three ways; (i) volume expansion promotes organ and tissue perfusion, (ii) the low viscosity of HBOC-201 improves flow to tissues, and (iii) oxygen delivery by HBOC Hb in the plasma is relatively insensitive to mechanisms regulating RBC distribution in the microcirculation. Diffusive oxygen transport is increased by the higher P50 compared with native RBC Hb which increases the off-loading of oxygen to tissues. Oxygen transport is also increased by reducing the diffusional barrier to oxygen transport associated with the plasma, in which oxygen is sparingly soluble. Biopure's HBOC solutions have been shown in vitro and in vivo to take up and off-load oxygen more efficiently than RBC Hb, and when added to blood can increase the efficiency of RBC oxygen transport. PMID- 14562724 TI - The concept of hemoglobin equivalency of perfluorochemical emulsions. AB - Perfluorochemical (PFC) emulsions have been in development as intravenous oxygen carriers for a number of years and many publications have dealt with their oxygen transport characteristics in both experimental models and in clinical trials. Though it has been stressed on numerous occasions that PFCs deliver oxygen to the tissues in very different ways to those by which Hemoglobin (Hb) releases oxygen (O2), no serious attempts have been made to correlate the oxygen delivery capacity of PFCs to those of Hb. This paper presents theoretical ways in which this can be done and demonstrates that a 2.7 g/kg dose of PFC is approximately equivalent to 4 g/dL [Hb]. Clinical trial planning is discussed. PMID- 14562725 TI - The dose-dependent effect of RSR13, a synthetic allosteric modifier of hemoglobin, on physiological parameters and brain tissue oxygenation in rats. AB - RSR13 is a synthetic allosteric modifier of hemoglobin that decreases the oxygen binding affinity of hemoglobin, increasing the P50. As a result, tissue oxygen tension is expected to increase. Using the capabilities of in vivo EPR, we directly examined the effect of RSR13 on brain pO2 in rats and the relationship between any change in brain oxygenation and changes in physiological parameters, including blood gases. The brain pO2 and arterial blood paO2 were increased significantly (p < 0.005) following RSR13 administration. The peak increase of brain tissue pO2 was 8.8 +/- 1.2 mm Hg in the animals receiving 150 mg/kg RSR13 and 13 +/- 3 mm Hg in the animals receiving 300 mg/kg RSR13. There was no difference among groups in MBP, heart rate, paCO2, pH, or HCO3. These data indicate that in anesthetized rats, RSR13 dose-dependently increases brain pO2 without affecting other physiologic parameters. This capability is likely to be very useful in circumstances where the pO2 of the brain is compromised. PMID- 14562726 TI - Computer modeling of relationship between critical PvO2, VO2max and blood supply of skeletal muscle at working with a right-shifted blood O2 dissociation curve. AB - Investigations were performed on a computer model of O2 delivery and O2 consumption in the one working muscle. At working with increasing power and achieving the critical value of VO2 (VO2crit), the muscle VO2 began to lag behind the oxygen demand qO2. The model permits to find critical pO2 in effluent venous blood Pvcrit at VO2crit as well as to calculate VO2max and PvO2 at VO2max under exercise with changing muscle blood flow F and blood pH.Pvcrit was computed from the condition VO2crit = 0.9qO2, and VO2max- from the condition (dVO2/dqO2) = 0.1. VO2max, Pv at VO2max, Pvcrit, and VO2crit were calculated for: 40 < or = F < or = 120 ml/min/100 g; 6.8 < or = pH < or = 7.4. It was shown that the faster is F and the lesser is blood pH, the greater were the Pvcrit and VO2max values. With decreasing F and blood pH, the influence of F on Pvcrit and VO2max increases, whereas the influence of blood pH on these values decreases. With increasing F and, hence, an increasing VO2max, the blood supply efficiency decreases due to the important limiting factor--tissue oxygen diffusion. PMID- 14562727 TI - Small-volume resuscitation with the hemoglobin substitute HBOC-201: effect on brain tissue oxygenation. AB - OBJECTIVES: To investigate the effects of small-volume resuscitation with a hemoglobin based oxygen carrier on brain tissue oxygen tension (PbrO2) in hemorrhaged swine. METHODS: Clark-type electrodes were inserted into the brain tissue of 6 swine to measure PbrO2 directly. Swine were hemorrhaged to a MAP of 40 mm Hg for 20 minutes. Resuscitation was performed with a bolus infusion of HBOC-201 (6 cc/kg; Biopure Corp.) and high-flow oxygen (100%). Swine were observed for an additional 2 hours. RESULTS: PbrO2 prior to hemorrhage was 48.7 +/- 4.7 mm Hg with 100% inspired oxygen. PbrO2 rapidly declined to 7.6 +/- 5.3 mm Hg in response to hemorrhage. Small-volume resuscitation with HBOC-201 and high flow oxygen resulted in a significant increase (p < 0.001) in PbrO2 to 44.6 +/- 8.1 mm Hg. MAP was also significantly increased to 84% of baseline. These elevations were sustained during the observation period. CONCLUSIONS: Resuscitation with HBOC-201 can restore and sustain cerebral oxygenation and MAP. These results suggest that a small-volume bolus of HBOC-201 may provide adequate oxygen and pressure support during the initial management of hemorrhage. PMID- 14562728 TI - The effect of RSR13, a synthetic allosteric modifier of hemoglobin, on brain tissue pO2 (measured by EPR oximetry) following severe hemorrhagic shock in rats. AB - RSR13 is a synthetic allosteric modifier of hemoglobin that decreases the oxygen binding affinity of hemoglobin, potentially increasing oxygen availability to hypoxic tissues. Using in vivo EPR to directly measure cortical pO2, we examined whether RSR13 would improve brain tissue pO2 following severe hemorrhagic shock in rats. Hemorrhagic shock was induced by withdrawing blood (2.7-2.8 mL/100 g/15 min). Following a 30 min shock period, resuscitation was performed by infusion with Ringer lactate plus RSR13 (150 mg/kg) or saline (control). Following hemorrhage, brain pO2 decreased by about 14 mm Hg in both groups. Following crystalloid resuscitation brain pO2 remained depressed in the control group but returned to the pre-hemorrhage values in the rats that received RSR13. RSR13 immediately increased and maintained the paO2 while controls had a very gradual increase towards pre-hemorrhage values. There was no difference in the blood pressure or heart rate between groups. RSR13 may have useful applications to decrease the effects of acute hemorrhagic hypoxemia by increasing brain oxygenation. PMID- 14562729 TI - Expression of myoglobin in the transgenic mouse brain. AB - The main purpose of this study was to express human myoglobin in mouse brain neurons and investigate the effects of this expression on metabolism and blood flow using phosphorous (31P) NMR spectroscopy and NMR perfusion imaging. Transgenic mice expressing brain myoglobin were created using a cDNA sequence for human myoglobin placed under the transcriptional control of either a human platelet-drived grown factor polypeptide B (PDGF-B) promoter sequence or a rat neuron-specific enolase (NSE) promoter sequence. The presence of myoglobin having a functional, reduced-state, heme group was demonstrated by protein analysis and immunocytochemistry. Expression levels were highest in the hippocampus, cerebellum, and cerebral cortex. No gross morphological adaptations of neural tissue resulting from the expression were observed and no statistically significant differences in the energetic state, as measured by 31P NMR, or baseline cortical perfusion, as measured by an NMR perfusion imaging technique, were found. PMID- 14562730 TI - The cerebral microcirculation in ischemia and hypoxemia. The Arisztid G. B. Kovach Memorial Lecture. AB - The cerebral capillary circulation exhibits heterogenous perfusion and undergoes characteristic changes in the distribution of RBC flow in response to systemic physiological stimuli. Hypoxemia, hypercapnia and hypotension increase the homogeneity of capillary perfusion, which is thought to preserve or enhance transcapillary exchange. Redistribution of capillary RBC flow between nutritive capillaries and preferential channels may contribute to this response. Selective changes in capillary flow may be brought about by non-smooth muscle-based contractile or blood-borne mechanisms. Isovolemic hemodilution anemia increases RBC velocity and supply rate with no decrease in capillary hematocrit. The effect of cerebral ischemia on microvascular patency depends on the severity and time course of the insult and whether the injury is global or focal. Capillary plugging is not observed following transient forebrain ischemia in the rat cerebral cortex but may contribute to tissue injury prior to reperfusion and during prolonged and severe ischemia. In the future, a better understanding of the functional architecture of the cerebral capillary network and its significance in the adaptation to altered circulatory conditions will continue to be an important goal of research. More work will have to be done to (i) substantiate the postulated physiological regulation of cerebral capillary flow, (ii) determine the cellular mechanism of integration of flow-dependent and neuronal activity-dependent signals, and (iii) identify the principal mediators, their cellular sources and molecular targets. The final answer to these questions will in a large part depend on our ability to directly, i.e. microscopically, visualize microvascular, neuronal and molecular phenomena as they occur in the brain in a spatially and temporally distributed manner. PMID- 14562731 TI - Investigating the role of nitric oxide in regulating blood flow and oxygen delivery from in vivo electrochemical measurements in eye and brain. AB - We have previously shown from direct, in vivo measurements of NO in cats with recessed electrochemical microsensors that NO mediates increases in ONH blood flow during functional activation of the eye by flickering light. We have also reported that there are low frequency (< 15 cycles/min) spontaneous oscillations in NO that appear to be passively coupled to oscillations in blood flow at similar frequencies in the cat ONH. In this paper, we describe similarities between in vivo measurements of NO in the ONH of the cat eye and in the cortex of the rat brain. These data are consistent with a role for NO in the coupling of blood flow with increases in neuronal activity, autoregulation of blood flow, hyperemia, and vasodilation during hypoxia and hypercapnia. PMID- 14562732 TI - NIRS monitoring of pilots subjected to +Gz acceleration and G-induced loss of consciousness (G-LOC). AB - With the increasing maneuverability of modern aircraft, there is an increased frequency of pilots losing consciousness due to high +Gz acceleration. This phenomena is defined as "G-induced loss of consciousness" (G-LOC). We used an NIRS system of our design to monitor cerebral oxygenation changes of pilots subjected to high +Gz acceleration and G-LOC. During the +Gz pulse, delta HbO2, and delta TotalHb decreased, with lesser changes of delta Hb. The maximum decrease of delta HbO2 and delta TotalHb usually occurred at the onset of G-LOC. After G-LOC, delta HbO2 and delta TotalHb increased rapidly for the first few seconds, beginning the reactive hyperemic recovery phase. delta HbO2 and delta TotalHb peaked, and then began to decrease towards baseline. The subjects were unconscious for 3-10 seconds after the onset of G-LOC. Upon returning to consciousness, the subjects were disoriented for another 4-11 seconds. NIRS provides an additional means of studying physiological mechanisms leading to and recovery from G-LOC. PMID- 14562733 TI - Correlation of NIRS determined cerebral oxygenation with severity of pilot +Gz acceleration symptoms. AB - Pilots commonly experience decreased peripheral vision, confusion & disorientation, and/or unconsciousness when exposed to high +Gz acceleration. We correlated NIRS determined delta Hb, delta HbO2, and delta TotalHb with the resultant +Gz stress symptoms that subjects reported after experiencing a 6 to 10 +Gz amplitude pulse. During the hyperemic response phase following the +Gz pulses, an increase of the averaged peak values of delta HbO2 and delta TotalHb as a function of the severity of the subjects' symptoms was observed. Significant increases were found for the averaged peak values of delta HbO2 and delta TotalHb between high vision loss, confusion and disorientation while remaining conscious (A-LOC), and unconsciousness (G-LOC). The results suggest that the confusion and disorientation associated with A-LOC is physiologically based and that A-LOC is an intermediate +Gz stress symptom between high peripheral vision loss and G-LOC. Like G-LOC, pilots who experience A-LOC symptoms momentarily do not have full control of their aircraft. PMID- 14562734 TI - Altered gene expression following cardiopulmonary bypass and circulatory arrest. AB - This study investigated the effects of normothermic cardiopulmonary bypass (CPB) and circulatory arrest (DHCA) on expression of specific genes in neonatal piglet brain. CPB was performed through the chest at 100 ml/kg/min for 2 hrs at 37 degrees C. In the second group of animals, CPB was begun as described above and then animals were cooled to a nasopharyngeal/brain temperature of 18 degrees C. When the brain temperature reached 18 degrees C, the CPB circuit was turned off. After 60 min of circulatory arrest (DHCA), CPB was resumed at 100 ml/kg/min, and the piglets were rewarmed to a temperature of 36 degrees C. In both groups, the animals remain sedated, paralyzed, mechanically ventilated, and continuously monitored throughout a four hour study period after CPB. Oxygen pressure in the microvasculature of the cortex was measured by oxygen dependent quenching of phosphorescence. The aRNA technique was used to assess mRNA steady-state levels in the brain tissue. Control oxygen pressure (pre-bypass) was 61 +/- 5 Torr and during CPB this decreased to 32 +/- 7 Torr on the beginning of bypass and to 36 +/- 5 Torr at the end of bypass. During the recovery period, cortical oxygenation steadily decreased, reaching 29 +/- 8 Torr at the end of the four hours period. Cortical oxygen decreased during DHCA to near zero and during rewarming and recovery increased to 35 +/- 6 Torr. Measurements of gene expression following CPB revealed significantly increased levels of mRNA for NMDAR1, DARPP-32, CamKII, GluR1, and D1AR. DHCA caused changes similar to those for CPB in levels of mRNA for NMDAR1, DARPP-32, CamKII and GluR1. In contrast, DHCA caused significantly increased levels of mRNA for GluR6 and GABRB1. There was no significant alteration in the level of D1AR following DHCA. The results showed that DHCA caused much larger alterations in gene expression in the critical metabolic signaling pathways tested than did CPB. PMID- 14562735 TI - Dominant events that modulate mass transfer coefficient of oxygen in cerebral cortex. AB - Recently, a model of cerebral oxygen delivery was described (J Appl Physiol 85:554) which yields a relationship similar to that used to depict substrate transport across the endothelium. Because the endothelium is not a diffusion barrier for oxygen, the permeability surface area product was replaced by an effective mass transfer coefficient term for oxygen, D. The cerebral metabolic rate of oxygen utilization (CMRO2) was linked to cerebral blood flow (CBF) and volume (CBV) through properties that modify the vessel-to-tissue oxygen tension giving rise to changes in D. Changes in the value of D were correlated with changes in CBF, CMRO2, and CBV as measured using NMR methods in a 48 microL volume of the cerebral cortex of anesthetized rats at different levels of activity. We conclude that the changes in total vascular volume (i.e., swelling or shrinking of the capillary bed) contributes < 5% to changes in D, whereas variations in the number of hematic vs. plasmatic capillaries, or intra-capillary stacking vs. unpacking of erythrocytes, or increase vs. decrease of dissolved oxygen in the tissue (i.e., processes which modify vessel-to-tissue oxygen tension) contribute(s) > 95% to changes in D. PMID- 14562736 TI - Relationship between redox behavior of brain cytochrome oxidase and neurological prognosis. AB - Currently, no on-line method of assessing cerebral oxygenation is sufficiently accurate to be clinically helpful. In an attempt to find a good predictor of postoperative cerebral outcome, we retrospectively studied the relationship between the redox behavior of cytochrome oxidase (cyt. ox.) during an operation and the neurological prognosis in 83 patients who underwent thoracic aortic surgery. Our data revealed three patterns of change in the redox behavior of cyt. ox. during the operation; the actual pattern exhibited by a given patient showed a highly significant correlation with the neurological prognosis (p < 0.0001). We conclude that the redox behavior of cyt. ox. during an operation is likely to be a good predictor of postoperative cerebral outcome, which implies that brain tissue oxygen sufficiency can be evaluated by near-infrared measurement of cytochrome oxidase (except for that in local regions far from the monitoring site). PMID- 14562737 TI - Are VEP correlated fast optical signals detectable in the human adult by non invasive nearinfrared spectroscopy (NIRS)? AB - The potential of near infrared spectroscopy (NIRS) to detect vascular changes in cerebral cortical tissue elicited by functional stimulation has been established (1). The vascular response is considered the result of neuronal activity in the investigated area and forms the basis of imaging techniques such as BOLD-contrast fMRI and PET. In the animal optical methods have been shown to detect optical changes in the illuminated tissue which exactly follow the time course of electrical potential changes, thus optical techniques can potentially assess both the 'fast' neuronal and the 'slow' vascular response. Lately a group of investigators has reported data showing that the 'fast optical signal' is detectable in the adult human in response to a visual stimulus ("optical VEP" termed EROS, i.e. event related optical signal). We failed to reproduce these results, with an almost identical instrumentation and experimental protocol. The negative result is reported in this paper. To check the expected magnitude of such changes we performed a simulation based on data reported in the animal and a human head model of optical properties. The result indicates that changes in optical properties to be expected in a non-invasive approach in the human adult are about three orders of magnitude smaller than those reported previously by the group of Gratton and co-workers (2). Also they are so small that they are below the noise level of the presently available NIRS monitors. PMID- 14562738 TI - Using high spectral and spatial resolution bold MRI to choose the optimal oxygenating treatment for individual cancer patients. AB - We evaluate whether high spectral and spatial resolution (HiSS) BOLD MRI can correctly rank the effects of three tumor-oxygenating treatments on radiosensitivity in BA1112 rhabdomyosarcomas (n = 5). Significant decreases in spectral linewidth predict that treatment with carbogen gas combined with a perfluorocarbon emulsion will increase radiosensitivity more than either treatment alone, which agrees with the known effects of these treatments on hypoxic fraction. High-resolution maps show that tumor response to each treatment is spatially heterogeneous, and that there is a paradoxical response to the treatments in 7-12% of tumor pixels. Because HiSS MRI emphasizes changes in necrotic and/or hemorrhagic regions, it is more sensitive to oxygenation changes compared to conventional MRI. These results demonstrate that HiSS MRI is a practical, noninvasive method that could be used to choose the treatment that maximizes the size and extent of increases in tumor oxygenation for individual patients. PMID- 14562739 TI - Issues in GRE & SE magnetic resonance imaging to probe tumor oxygenation. AB - Tumor oxygenation determines the efficacy of radiotherapy, but there is no non invasive way to image this parameter. Since gradient recalled echo (GRE) images are sensitive to blood deoxyhaemoglobin concentration ([dHb]) they could have a role in assessing tumor oxygenation. In brain, linear relationships have been demonstrated between brain tissue R2* relaxation rate and tissue [dHb] or oxygen saturation, but in tumors, vascular and tissue heterogeneity, and the presence of simultaneous oxidative and glycolytic metabolism, complicate the analysis. We have studied the effects of vascular challenge in a rat prolactinoma tumor model by MR imaging and spectroscopy and comment on the implications of these results for calibrating GRE images for blood or tissue pO2. PMID- 14562740 TI - fMRI for monitoring dynamic changes in tissue oxygenation/blood flow: potential applications for tumor response to carbogen treatment. AB - The ability to differentiate between well-oxygenated and poorly-oxygenated tumors may play an important role in selecting an optimal therapeutic regime for tumor treatment of the individual patient. We present preliminary results in the development of a dynamic functional MRI method for mapping tissue oxygenation and blood flow distribution in humans simultaneously. We applied interleaved Blood Oxygenation Level Dependent (BOLD) and Flow-sensitive Alternating Inversion Recovery (FAIR) sequences to detect signals as a subject is inspiring gases of varying oxygen concentration. The method allows quantitation of the spatial distribution and time course of the important physiological functions that are easily registered with high resolution anatomic MR images. It may be used to critically evaluate the efficacy of varying durations of carbogen breathing in tumor patients, and allow a quantitative evaluation of the roles of carbogen and other radiosensitizers as potential adjuncts to radiotherapy and drug therapies. PMID- 14562741 TI - Comparison study of oxygen-induced MRI-signal changes and pO2 changes in murine tumors. AB - The purpose of this study was to compare the results from oxygen-induced MR signal intensity changes with polarographic pO2 measurements in tumors. Balb-c mice with an intramuscular transplanted osteosarcoma were examined. To study the response of tumors to changes in oxygen supply, hyperoxia was induced by breathing pure oxygen for a short period. The examination of each animal started with T2* weighted MRI followed by the pO2 measurements (Eppendorf Histograph). During oxygen inhalation in all tumors, when the hypoxic tumor fraction drops, both areas of significant MR-signal intensity increase and decrease were observed in each animal. PMID- 14562742 TI - Venous-arteriolar reflex in human gastrocnemius studied by NIRS. AB - Heat-up tilting manoeuvre from 0 to 60 degrees induces oxygenated and deoxygenated haemoglobin concentration changes in the human gastrocnemius. These changes, measured by NIRS, can only be partially explained by the blood volume displacement due to the gravitational force. In the present study it is demonstrated, by a dye dilution technique (indocyanine green), that a reduction in blood flow (venous-arteriolar and/or spinal reflex) is responsible of the limited oxyhaemoglobin concentration increase observed when going from 0 (2.54 +/ 0.48 blood flow in arbitrary units, a.u.) to 60 (1.46 +/- 0.55 a.u.) degrees. The proposed technique is potentially applicable to the detection of specific pathological aspects of microcirculation, such as arterial occlusion in the leg, diabetes mellitus, and congestive heart failure, where the venous-arteriolar reflex may be affected. PMID- 14562744 TI - Modeling of oxygen diffusion from the blood vessels to intracellular organelles. AB - We describe recent models of oxygen transport in tissue along the pathway from the hemoglobin molecule to the mitochondria and illustrate their applications. Microvasculature is the major site of exchange between blood and parenchymal cells for gases (O2, CO2, CO, NO), nutrients, metabolic products, and drugs. These exchange processes are affected by the architecture of the microvessels and the surrounding cells; distribution of blood flow; transport characteristics of blood, cells, and interstitial space; and rates of various chemical reactions associated with the transport processes. These processes operate at multiple levels of biological organization, from the molecular to the organ levels. Quantitative understanding of molecular transport in cells and tissues, specifically of oxygen transport, is the prerequisite for understanding the mechanisms of many diseases and for designing effective therapies. Mathematical and computational models provide a powerful set of tools for studies of these complex phenomena. PMID- 14562743 TI - Muscle oxygen consumption at onset of exercise by near infrared spectroscopy in humans. AB - In this study, we tried to continuously measure muscle oxygen consumption (m-VO2) by near infrared spectroscopy (NIRS) without arterial occlusions. We used an intermittent isometric exercise at high intensity, which elicits a spontaneous occlusion of the blood flow to the muscle due to an increase in intramuscular pressure. Changes in muscle oxygenation and phosphocreatine (PCr) concentration were monitored in 5 subjects during an intermittent isometric exercise (5 sec. contraction/5 sec. relaxation) at 50% of maximum voluntary contraction for 3 minutes. The rate of deoxygenation was measured from the 2nd sec. to the 3rd sec. of each muscle contraction. The rate of deoxygenation at the onset of exercise followed an exponential time course with a time constant of 42.0 +/- 12.5 sec. (mean +/- SD). This value agreed with the time constant of the decrease in PCr (48.2 +/- 10.2 sec.). This result suggests that m-VO2 was successfully monitored with a time resolution of 10 sec. by NIRS during exercise without arterial occlusion. PMID- 14562745 TI - Muscle reoxygenation rate after isometric exercise at various intensities in relation to muscle oxidative capacity. AB - The purpose of this study was to determine whether the reoxygenation rate (Reoxy rate) immediately after static exercise at various submaximal intensities would be related to muscle oxidative capacity. Seven healthy male subjects performed isometric handgrip exercise for 10 sec at 30%, 60% and 90% of maximal voluntary contraction (MVC). The Reoxy-rate and muscle oxygen consumption during exercise (muscle VO2EX) were monitored by near infrared continuous wave spectroscopy (NIRcws). The muscle oxidative capacity was evaluated by the time constant for phosphocreatine resynthesis (PCrTc) using 31-phosphorus magnetic resonance spectroscopy (31P-MRS). The Peak blood flow of brachial artery after exercise (BABFpeak) was measured using Doppler ultrasound. There was no correlation between PCrTc and Reoxy-rate at 30% and 60% MVC. In contrast, Reoxy-rate at 90% MVC was positively correlated to PCrTC (r = 0.825, p < 0.05). The muscle VO2EX increased 5.9, 8.8 and 12.6-fold of the resting on average at 30%, 60% and 90% MVC, respectively, and the muscle VO2EX at 90% MVC was significantly higher than that at 30% and 60% MVC. On the other hand, BABFpeak increased only just 1.9, 2.4 and 2.7-fold of the resting on average at 30%, 60% and 90% MVC, respectively (Fig. 4). These results suggest that the higher oxidative capacity muscle shows slower muscle reoxygenation after 10 sec isometric exercise at 90% MVC because the Reoxy-rate after this type of exercise may be influenced more by muscle VO2 than by O2 supply. In contrast, 60% MVC and lower exercise intensities may not be severe enough to influence the muscle VO2 dependent Reoxy-rate. PMID- 14562746 TI - Oxygen diffusion coefficient and oxygen permeability of metmyoglobin solutions determined in a diffusion chamber using a non-steady state method. AB - Non-steady state measurements of oxygen diffusion through various model layers can be performed using a diffusion chamber that was described earlier [1, 2]. A closer analysis of these measurements showed that they not only yield the oxygen diffusion coefficient (DO2) of the diffusion layer, but also the oxygen permeability (PO2). In this study DO2 and PO2 have been determined in solutions of metmyoglobin (metMb) with concentrations varying between 5 and 40 g/dL at 25 degrees C. Both DO2 and PO2 decreased with increasing metMb concentration. This decrease was comparable to the values reported for DO2 in protein solutions by Kreuzer and Hoofd [3]. Using this diffusion chamber for non-steady state measurements, oxygen diffusion coefficients, oxygen permeability and oxygen solubility (aO2 = PO2/DO2) of various model layers could be determined. PMID- 14562747 TI - Pinacidil-induced opening, like glibenclamide-induced closure of cardiac KATP channels, protects cardiac function against ischemia in isolated, working, erythrocyte perfused rat hearts. AB - Glibenclamide-induced closure of ATP-dependent potassium (KATP) channels decreases coronary blood flow during normoxic and post-ischemic conditions. We have found that post-ischemic cardiac function is improved after glibenclamide treatment. Our theory was that this is a result of higher intracellular calcium concentrations due to reduction in ischemia-mediated hyperpolarization of the myocardial cell membrane. We hypothesized therefore that opening KATP channels would reduce post-ischemic function in our isolated, erythrocyte perfused, working rat heart model. During treatment with 1 or 12 mumol.L-1 pinacidil (protein unbound concentration) both before and after 12 minutes global ischemia coronary blood flow increased 2-3 fold compared with vehicle, while cardiac functional recovery post-ischemically was improved with both concentrations. Because closing and opening cardiac KATP channels both improve post-ischemic function, our calcium theory above can be discounted. The protective effect of glibenclamide may possibly be ascribed to metabolic effects such as preservation of ATP levels during ischemia. PMID- 14562748 TI - Effect of irradiation on enzymes of the capillary bed in rat ventricles. AB - The effect of localized irradiation on the enzyme activity in rat cardiac capillaries was examined in experiments in which the arteriolar and venular portions of the capillary bed were distinguished by the double-staining method. This method shows that the endothelial cells of the former contain alkaline phosphatase (AP) and those of the latter, dipeptidylpeptidase IV (DPP). At both 1 week and 3 weeks after irradiation with 20 Gy, staining for AP was reduced but staining for DPP was unchanged. The loss of enzyme from the arteriolar portions may be a consequence of the greater radiosensitivity of tissues exposed to high oxygen tension, or it may indicate that AP is less stable than DPP when exposed to irradiation. PMID- 14562749 TI - The redox state of cytochrome oxidase in brain in vivo: an historical perspective. AB - Recent evidence suggests that cytochrome oxidase is partially reduced under resting conditions in the brain. Previous data, recorded over the past 30 years from intact brain using optical methods in the visible wavelength range, are consistent with this observation. These older data, while not conclusive in themselves, support the overall conclusions. The historical perspective on the experiments and controversies illustrates a number of useful principles. The first is that new methods tend to produce new observations that may be difficult to reproduce due to the uniqueness of the instrumentation. The second is that any new and different observations cannot be assimilated without an acceptable theoretical framework and, without assimilation can have little impact. Finally, the mechanisms which might explain why cytochrome oxidase may be more reduced than previously thought are still not fully developed and, therefore, the physiological significance of such reduction is not known. PMID- 14562750 TI - A quantitative study of oxygen as a metabolic regulator. AB - An acute reduction in oxygen delivery to a tissue is associated with metabolic changes aimed at maintaining ATP homeostasis. However, given the complexity of the human bioenergetic system, it is difficult to determine quantitatively how cellular metabolic processes interact to maintain ATP homeostasis during stress (e.g., hypoxia, ischemia, and exercise). In particular, we are interested in determining mechanisms relating cellular oxygen concentration to observed metabolic responses at the cellular, tissue, organ, and whole body levels and in quantifying how changes in tissue oxygen availability affect the pathways of ATP synthesis and the metabolites that control these pathways. In this study, we extend a previously developed mathematical model of human bioenergetics, to provide a physicochemical framework that permits quantitative understanding of oxygen as a metabolic regulator. Specifically, the enhancement--sensitivity analysis--permits studying the effects of variations in tissue oxygenation and parameters controlling cellular respiration on glycolysis, lactate production, and pyruvate oxidation. The analysis can distinguish between parameters that must be determined accurately and those that require less precision, based on their effects on model predictions. This capability may prove to be important in optimizing experimental design, thus reducing use of animals. PMID- 14562751 TI - The oxygen dependency of cerebral oxidative metabolism in the newborn piglet studied with 31P NMRS and NIRS. AB - Mean cerebral saturation and changes in the oxidation state of the CuA centre of cytochrome oxidase were measured by near infra-red spectroscopy simultaneously with phosphorous metabolites and intracellular pH measured using 31P NMR spectroscopy during transient anoxia (inspired oxygen fraction = 0.0 for 105 seconds) in the newborn piglet brain. By collecting high quality 31P spectra every 10 seconds, it was possible to resolve the delay between the onset of anoxia and the fall in PCr and to show that the CuA centre of cytochrome oxidase reduced simultaneously with the fall in PCr. From these observations it is concluded that, at normoxia, oxygen tension at the mitochondrial level is substantially above a critical value at which oxidative metabolism becomes oxygen dependent. PMID- 14562752 TI - Effect of myoglobin inactivation on intracellular gradients of NADH fluorescence at critical mitochondrial oxygen supply. PMID- 14562753 TI - The effects of anesthesia on cerebral tissue oxygen tension: use of EPR oximetry to make repeated measurements. AB - While very useful data can be obtained from measurements of pO2 within various compartments of the vascular system, such measurements do not necessarily provide accurate information on the pO2 in the brain. Anesthetics can significantly affect the tissue pO2 in the brain by several mechanisms involving both delivery and utilization. Electron Paramagnetic Resonance (EPR or ESR) oximetry has the potential for non-invasively carrying out repeated direct measurements of pO2 in tissues during the course of anesthesia. In this paper we describe the use of EPR oximetry for studying the influence of anesthesia on tissue pO2, and present illustrative results from experiments with five different anesthetics in rats. The results indicate that the tissue O2 can be measured directly using EPR oximetry, and data can be obtained non-invasively during the course of anesthesia. PMID- 14562754 TI - Capillarization and vascular endothelial growth factor expression in hypertrophying anterior latissimus dorsi muscle of the Japanese quail. AB - Hypertrophy may increase the diffusion distances from capillaries to the interior of the muscle fibers. We hypothesized that capillary proliferation occurs during hypertrophy, which is accompanied by an up-regulation of vascular endothelial growth factor (VEGF). Hypertrophy of the left anterior latissimus dorsi muscle of Japanese quail (2-3 months old) was induced by 1-4 week stretch-overload. Capillarization was analyzed in cross-sections stained for ATPase. VEGF expression was determined with RT-PCR. Initially, hypertrophy was not accompanied by increases in fiber cross-sectional area (FCSA), but after 1 week the average FCSA did increase. The capillary to fiber ratio was decreased after 1 week, but returned to control values in subsequent weeks. This indicates that capillary proliferation occurred, because this model is characterized by extensive fiber proliferation. The absence of any significant change in VEGF mRNA levels indicates that increased levels of VEGF mRNA are not crucial for capillary proliferation during muscle hypertrophy. PMID- 14562755 TI - Expression of proliferating cell nuclear antigen in rat hearts subjected to transient ischemia followed by reperfusion. AB - Early mechanisms involved in improving capillarity and oxygen transport to cardiac tissue exposed to transient coronary ischemia followed by reperfusion were studied in rats. Under ether anaesthesia, the left coronary artery was mechanically occluded for 3 min after which it was released, and the rats allowed to recover. After 2, 24 or 48 h the rats were sacrificed and the hearts frozen in liquid nitrogen. Frozen cross-sections were stained immunohistochemically for proliferating cell nuclear antigen (PCNA) and for the growth factors, VEGF and bFGF. No reaction for PCNA was seen in sections of sham-operated hearts but an inhomogeneous reaction occurred in annular structures in the occluded hearts at 48 h reperfusion. The stain appeared to be located in proliferating nuclei, and in the cytosol of endothelial cells. It is suggested that PCNA is stimulated by the increase in growth factors that is known to occur within 2 h after the end of the coronary occlusion. It is concluded that the increase in capillarity, indicated by the nuclear proliferation of endothelial cells, will improve the transport of oxygen to the cardiac tissues. PMID- 14562756 TI - Fibroblast growth factors (FGFS) increase breast tumor growth rate, metastases, blood flow, and oxygenation without significant change in vascular density. AB - Breast tumors expressing no detectable FGFs (MCF-7) were compared with tumors transfected with FGF4 or FGF1 (FGF4/MCF-7 or FGF1/MCF-7), and with MDA-MB-435, which produce endogenous FGF2. Tumor blood flow was measured by 133Xe diffusion, oxygen distribution was measured by Eppendorf pO2 histography, and vascular density was measured by CD31 staining. Tumors that overexpress angiogenic factors grew at a rate far exceeding that of MCF-7. The FGF producing tumors also had much higher metastatic rates to lung. Tumor blood flow was significantly higher in the two FGF-transfected xenografts compared with the parent MCF7. Median tumor pO2 was also higher, and tumor oxygenation was preserved even for large tumors. The vascular density as determined by CD31 staining, however, was not markedly increased in tumors overexpressing angiogenic factors. We found that angiogenic factors preserve and augment neovascular function, thus facilitating tumor growth and progression. PMID- 14562757 TI - FGF1 and VEGF mediated angiogenesis in KHT tumor-bearing mice. AB - Isotransplants of murine fibrosarcoma (KHT) cells were inoculated i.m. into the hind limbs of 6-8 week-old female C3H/HeJ mice. Intratumoral injection of FGF1 or VEGF proteins decreased hypoxic marker uptake in murine fibrosarcoma KHT. Reduction of tumor hypoxia did not correlate with mRNA expression of transcription factors in tumors. Likewise, there was no significant alteration in either apoptotic frequency or the mRNA levels of 10 apoptotic-related molecules in FGF1- or VEGF-treated tumors. mRNA expression for MCP-1, IL-1 beta, IL-18, and IL-1Ra, however, were decreased in the tumors following FGF1 or VEGF treatment. Among the normal tissues tested (brain, kidney, liver, spleen, and lung), basal mRNA levels for cytokines and chemokines varied. Intratumoral injection of FGF1 or VEGF (6 daily intra-tumor injections of 6 micrograms/mouse) did not alter most cytokine or chemokine mRNA expression in spleen and lung. In summary, alteration of tumor oxygenation by local administration of angiogenic growth factors may be mediated by cytokine/chemokine production in the tumor. PMID- 14562758 TI - HIF-1 alpha and VEGF expression after transient global cerebral ischemia. AB - Hypoxia inducible factor-1 alpha (HIF-1 alpha) and vascular endothelial growth factor (VEGF) expression were studied in rat cerebral cortex after reversible global cerebral ischemia produced by cardiac arrest and resuscitation. Immunoblot analysis showed a significant induction of HIF-1 alpha protein after 1 hour of recovery from cardiac arrest which remained elevated for at least 12 hours. Upregulation of VEGF mRNA and protein were also observed but this was delayed in comparison to the HIF-1 alpha response. VEGF188 and VEGF164 mRNA levels were increased at 12-48 h of recovery from cardiac arrest but returned to basal expression after 7 days. Changes in VEGF120 mRNA expression did not reach statistical significance. Correspondingly, VEGF protein levels increased by about double at 24 and 48 hours of recovery but returned to basal levels after 7 days. These results suggest that cardiac arrest and resuscitation triggers HIF-1 alpha induction, which might be at least in part responsible for the stimulation of VEGF expression. PMID- 14562759 TI - Relationship between the gene expression of c-fos and degree of hypoxia in rat brain, as revealed by near-infrared spectroscopy. AB - Hypoxic induction of c-fos was studied in rat brains as a function of the cerebral oxygenation state using near-infrared spectroscopy by which the hemoglobin oxygenation state and redox state of mitochondrial cytochrome oxidase can be monitored noninvasively. Following reoxygenation after hypoxia, the expression of c-fos and MAP2 mRNAs was determined by reverse transcription coupled PCR. The expression of MAP2 remained unchanged throughout all conditions from 21 to 8% FiO2. Under the mildly hypoxic conditions, c-fos mRNA was not induced. Hemoglobin was partially deoxygenated but cytochrome oxidase remained fully oxidized. Severe hypoxia, where cytochrome oxidase was reduced, caused a significant induction of c-fos mRNA. At this stage, the oxygen concentration in cerebral tissue fell to lower than 10(-7) M. These data suggest that the decline in oxidative phosphorylation might be a trigger for the induction of c-fos mRNA. PMID- 14562760 TI - Blood volume changes are controlled centrally not locally--a near-infrared spectroscopy study of one legged aerobic exercise. AB - It is well known that blood flow increases in an exercising limb to match increases in oxygen consumption. However, it is less clear what effects occur in the opposite limb. We performed a one legged incremental cycling protocol and used near-infrared spectroscopy to measure changes in muscle blood volume and oxygenation in the exercising and non-exercising leg. As expected during exercise the exercising leg was deoxygenated relative to the non-exercising leg. However, there were similar increases in blood volume to both legs during the exercise, and during the post-exercise recovery period similar volume and oxygenation increases were seen in both legs. We conclude that blood volume increases may be signalled locally, but the effect is expressed globally. Previous studies have demonstrated a training effect in the non-exercising leg following one legged aerobic exercise. The large haemodynamic changes in the non-exercising leg observed here may be partially responsible for the cross-training effect. PMID- 14562761 TI - A modeling investigation to the possible role of myoglobin in human muscle in near infrared spectroscopy (NIRS) measurements. AB - Near Infrared Spectroscopy (NIRS) analyzes infrared light having traveled through tissue, for its oxygenation status. The main chromophore analyzed is hemoglobin (Hb), but in muscle tissue also myoglobin (Mb) is present. Since NIRS cannot discern between these two species experimentally, we did model calculation studies using general data for human muscle. Where such data were not directly available, we derived these from analogous data or straightforward assumptions. Consequently, conclusions have to be drawn cautiously. Solid conclusions are, that myoglobin is an important factor with red muscle, and that it is always partly desaturated, significantly depending on workload. Here, both deoxygenated Hb and Mb as detected by NIRS varied between 0.04 and 0.13 mol/m3, while the variation in Mb saturation (53-86%) even exceeded that of Hb (63-84%). PMID- 14562763 TI - Effects of the contrast medium iopromide on renal hemodynamics and oxygen tension in the diabetic rat kidney. AB - We investigated the effects of the contrast medium (CM) iopromide on regional renal blood flow and oxygen tension (pO2) in the streptozotocin (STZ)-induced diabetic Wistar Furth rats. RESULTS: In normoglycemic rats, CM injection induced a transient decrease followed by an increase in renal cortical blood flow (CBF), whereas CBF increased directly in the diabetic animals. Renal outer medullary blood flow (OMBF) increased in controls, while it decreased in the diabetic animals following CM injection. In control rats a marked initial decrease in OM pO2 following injection of CM was observed. In animals diabetic for 4 weeks only a slight decrease was seen, whereas in 9-week diabetic animals a persistent increase was recorded. CONCLUSIONS: An altered oxygen tension and hemodynamic response to CM was found in diabetic rats. If these disturbances may contribute to the development of renal dysfunction by CM in the diabetic rat kidney remains to be elucidated. PMID- 14562762 TI - Tissue hypoxia during bacterial sepsis is attenuated by PR-39, an antibacterial peptide. AB - Endotoxin (a lipopolysaccharide (LPS) component of the Gram negative bacterial cell wall) induces sepsis in laboratory animals and is the cause of septic shock in patients. Tissues often develop necrotic regions, particularly in kidney and liver, thought to be directly the result of endotoxin-induced release of nitric oxide (NO). These studies investigated the potential of PR-39, an antibacterial peptide, as an alternative treatment for sepsis. Our rationale for these experiments was based on the knowledge that PR-39 inhibits the superoxide producing NADH/NADPH-oxidase system, and also inhibits NOS. In a mouse model of sepsis, we carried out EPR measurements of liver pO2 and NO simultaneously in vivo. Physiological parameters were also measured in these animals (blood pressure, heart rate). NO levels in blood were measured by EPR analysis of red blood cell nitrosyl-hemoglobin. We found PR-39 alleviated endotoxin-induced liver hypoxia 6 hrs after treatment. Tissue NO was higher in the PR-39 + LPS group compared to LPS alone. Circulating levels of NO were the same in these groups. Taken together, these results suggest PR-39 is effective in improving survival following a septic episode. The exact mechanism is unclear, but increased NO as a result of decreased superoxide production seems to play an important role in alleviating tissue hypoxia. PMID- 14562764 TI - Postocclusive reactive hyperemia in healthy volunteers and patients with peripheral vascular disease measured by three noninvasive methods. AB - Postocclusive reactive hyperemia (PORH) was evaluated in three healthy volunteers and in three patients with different etiologies and suffering from peripheral arterial occlusive disease (PAOD). Three noninvasive methods were used: transcutaneous oximetry (TcPO2), near-infrared spectroscopy (NIRS), and laser Doppler flowmetry (LDF). Changes in perfusion and oxygenation of tissue were measured on foot before, during, and after arterial occlusion on thigh. Numerical parameters were derived from measured signals for quantification of the PORH response. Results of all three methods provided distinction between healthy volunteers and patients. The experimental optical techniques of NIRS and LDF demonstrated more clearly than the well-established TcPO2 method the difference between healthy volunteers and patients. The dynamics of the PORH response proved to be a better indicator of peripheral vascular disorder than the amplitude of responses. PMID- 14562765 TI - Role of myoglobin in regulating respiration. AB - The 1H NMR Val E11 signal provides a unique opportunity to observe carbon monoxide (CO) inhibition of Mb in the in vivo myocardium and to assess the functional role of Mb in regulating respiration. Upon carbon monoxide infusion, the MbO2 Val E11 signal at -2.76 ppm gradually disappears, and a new signal at 2.26 ppm, corresponding to MbCO, emerges. These signals yield the intracellular partial pressure of both O2 and CO and the extent of Mb inactivation, since CO binds more tightly to Mb than O2. Although contractile function decreases slightly to a steady state level, it shows no dose dependence on pCO. Up to 80% MbCO saturation, the contractile function remains at the steady state level. Neither the PCr concentration nor the oxygen consumption rate is significantly perturbed. Above 80% MbCO saturation, the oxygen consumption rate starts to decline. The experimental observations raise provocative questions about the functional role of Mb in the cell. PMID- 14562766 TI - Oxidative defenses in the sea bass, Dicentrarchus labrax. AB - A study on the oxidative defenses during larval growth and under stress conditions was carried out in the bred sea bass, Dicentrarchus labrax. A high pressure liquid chromatography (HPLC) method was used for the quantitative determination of vitamins C and E; glutathione peroxidase activity was measured by an enzymatic assay. Vitamin E was measured in the seminal fluid, eggs, embryos and larvae. Vitamins C and E, and glutathione peroxidase activity were measured in adults of Dicentrarchus labrax under normal conditions and subjected to hypoxia. Vitamin E content was high in seminal fluid, eggs, and embryos and at the early stage of larval development. It decreased slowly, but steadily, throughout the larval growth. In adults exposed to hypoxia, vitamins C and E levels were significantly lower with respect to the control group. Glutathione peroxidase levels showed a decrease in the hypoxia-subjected group, although the values were not significant. PMID- 14562767 TI - Rheologic dissimilarities in female and male blood: potential link to development of cardiovascular diseases. AB - Oxygen Delivery Index (ODI) was introduced as the ratio of red blood cell concentration (hematocrit) to blood viscosity. The ODI can be considered an indirect characterization of oxygen transport to organs and tissues. ODI was obtained for 98 healthy donors (47 pre-menopausal women and 51 age-matched men). In this population ODI levels were found to be significantly lower (p < 0.001) in male blood (7.7 +/- 0.3 vs. 8.4 +/- 0.5 in female blood). Average ODI obtained for 15 cardiac patients (all males) was found to be significantly lower than that for healthy men. In red blood cell suspensions with the same hematocrit, ODI was found to decrease when plasma viscosity was increased via an increase in protein concentration. Additionally, it was found that ODI measured for samples of blood over a wide hematocrit range, obtained by dilution with autologous plasma, possessed the highest values at the hematocrit levels 30 to 40%. The decreased oxygen transport might contribute to the significantly higher morbidity and mortality from cardiovascular diseases for men compared to pre-menopausal women. ODI may be a useful parameter for evaluation of risk of development of cardiovascular disorders. PMID- 14562768 TI - Preliminary studies of the application of near infrared spectroscopy in the diagnosis of deep vein thrombosis. AB - A feasibility study for diagnosing deep vein thrombosis utilizing near-infrared continuous wave spectroscopy was performed, as a real-time, non-invasive, and inexpensive method. The probe contains two light sources and two detectors with optical filters that monitor reflected light at wavelengths 760 and 850 nm to measure the changes in the amount of deoxyhemoglobin and oxyhemoglobin, respectively. These changes and the blood volume changes are recorded while the subject performs a series of light leg exercises. The test protocol is designed to determine the muscle tissue blood volume capacity, rate of blood filling (venous valve functionality test), and efficiency to promote one-directional venous flow from the leg to heart. The subject pool consists of the patients with leg deep vein thrombosis (DVT) diagnosed by the Johns Hopkins Hospital Vascular Surgery Department and of normal subjects as the control. Abnormal venous systems showed distinct characteristics: high blood volume in the leg; high rate of blood filling while the subject stands upright; and the inability to decrease the blood volume during the muscle contraction. The NIR device proved to be an inexpensive, effective, and portable device that can detect DVT in the leg in real-time. PMID- 14562769 TI - Cerebrovascular response to acute metabolic acidosis in humans. AB - OBJECTIVES: Evaluation of the cerebrovascular response (delta CBV/delta PaCO2) during baseline metabolic conditions and acute metabolic acidosis. METHODS: 15 healthy subjects, 5 m, 10 f, 56 +/- 10 yrs were investigated. For acidification, NH4Cl was given orally. CBV was measured using Near Infrared Spectroscopy (OXYMON) during normo-, hyper- and hypocapnia. RESULTS: Acute metabolic acidosis was realised: mean delta BE -2.7 mEq.L-1 (p < 0.001) with mean delta PaCO2 -0.2 kPa (p < 0.01). During normo-, hyper- and hypocapnia, CBV values of 3.51, 4.82 and 2.55 mL.100 g-1 were calculated during baseline metabolic conditions and 3.70, 4.86 and 2.63 mL.100 g-1 during acute metabolic acidosis. The CBV/PaCO2 response showed a hockeystick configuration with the point of infliction around normocapnia. delta CBV/delta PaCO2 reactivity from normo- to hypercapnia and from normo- to hypocapnia was calculated; no significant differences in delta CBV/delta PaCO2 were found in both metabolic conditions. CONCLUSION: Cerebrovascular reactivity to CO2 does not alter during acute metabolic acidosis. PMID- 14562770 TI - Free flap monitoring in plastic and reconstructive surgery. AB - Free flaps are regularly used in plastic and reconstructive surgery but have a significant failure rate due to vessel thrombosis in the re-established arterial or venous circulation. A monitor of flap perfusion and oxygenation would allow the early detection of progressing flap ischaemia, hastening the required intervention and maximising the chances of salvaging the flap. A dual wavelength spectrophotometer has been designed and constructed which can monitor haemodynamic events in flaps during surgery and postoperatively. Eleven patients undergoing free flap surgery were studied. Measurements were made during surgery before division of the vessels and during and after microvascular anastomosis. Significant changes in all parameters were observed on reperfusion of the flaps after anastomosis or tourniquet ischaemia. Abnormal reperfusion in one flap and subsequent ischaemic events in two others were identified. PMID- 14562771 TI - Inhibition of mitochondrial respiration during early stage sepsis. AB - It is known that nitric oxide (NO) is produced in response to a septic insult such as bacterial invasion and that overproduction of NO can have serious debilitating consequences. The mechanism by which NO causes damage at the cellular level is less clear. We have therefore studied the response to a septic insult in an anaesthetised spontaneously breathing Sprague-Dawley rat model. Six rats were given either an intravenous infusion of bacterial cell wall lipopolysaccharide (LPS, 5 mg/kg) or saline control over 1 hour. For electron paramagnetic resonance (EPR) studies, blood samples were collected every hour for a further two hours and liver tissue samples were collected postmortem. Measurement was also made of PaO2, blood pressure, base deficit, aortic and renal blood flow and hepatic microvascular pO2 (using porphyrin phosphoresence). Tissue samples were also collected for mitochondrial complex activity analysis. After the administration of LPS blood pressure, blood flow and microvascular PO2 were diminished and the base deficit increased. In addition a clear difference was observed by EPR between control and insulted blood and tissue samples. A large heam-nitrosyl signal is observed as well as an increase in the signal at g = 1.94, corresponding to the iron-sulphur centres of complex I becoming more reduced. However, no significant difference was observed for any of the mitochondrial complex activities. The effect of the NO produced was to depress the circulatory variables and increase base deficit, combined with a reduced oxygen consumption this implies an impairment of normal aerobic respiration. This was supported by increased iron-sulphur signals observed by EPR indicating a blockage in the mitochondrial redox chain with the subsequent accumulation of electrons. As no effect was observed in the mitochondrial complex activities this indicates that this inhibition is reversible in early stage sepsis. We conclude that nitric oxide produced in response to a septic insult can inhibit mitochondria causing an impairment of oxygen utilisation by aerobic respiration. PMID- 14562772 TI - [Rheumatology]. PMID- 14562773 TI - [What information is given on adverse drug reactions from new drugs?]. AB - BACKGROUND: It is not known to what extent information on previously unknown adverse reactions is communicated to doctors after the approval of a new drug. MATERIAL AND METHODS: We included seven drugs, approved between 1982 and 1995 and the first in their class to be approved in Norway. We recorded the number of adverse reactions listed in the annual editions of the Norwegian physicians' desk reference(Felleskatalogen) from the first edition that included the drug and up until the 2001 edition. RESULTS: Only 51 % of the adverse reactions listed in the 2001 edition were included in the first post-approval edition. On average, 1.6 new adverse reactions were added for each drug every year. By contrast: in a group of 12 drugs approved before 1970, 0.14 new adverse reactions were added for each drug every year between 1989 and 2001. INTERPRETATION: With a new drug it is to be expected that many adverse reactions are not acknowledged as such. Caution is warranted because of the fact that even though it is not listed in the physicians' desk reference, any event appearing in a patient exposed to a new drug may be an adverse reaction. PMID- 14562774 TI - [Diagnosis of cryptic chromosome aberrations]. AB - BACKGROUND: Cryptic chromosome aberrations, i.e. those that are not observed by routine chromosome analysis (G-banding), can cause a plethora of developmental abnormalities, usually at least a combination of various dysmorphic signs and mental retardation. MATERIALS AND METHODS: We present the current molecular cytogenetic methods for detection of cryptic chromosome aberrations, with special emphasis on comparative genomic hybridisation (CGH), a DNA-based screening method. RESULTS: Using comparative genomic hybridisation with high-resolution analysis, we were able to detect a chromosome aberration in 10 % of patients found to have normal karyotypes by standard chromosome analysis. INTERPRETATION: Though the sensitivity of comparative genomic hybridisation is still insufficient for finding a deletion in most of the well-known microdeletion syndromes, the diagnostic yield of genomic imbalances is better than for all other laboratory investigations except routine G-banding. PMID- 14562775 TI - [Secondary anterior sphincteroplasty for anal incontinence after delivery]. AB - BACKGROUND: The prevalence of anal sphincter rupture at vaginal delivery is from 0.5 to 2.5 %. More than 50 % of these patients may need a secondary anterior sphincteroplasty because of anal incontinence. MATERIALS AND METHODS: We present prospective series of 29 women (median age 45 years) operated with an overlapping anterior sphincteroplasty from 1996 to 2001. The patients were examined with anal ultrasonography (n = 23), manometry (n = 19) and neurophysiologically (n = 13), and divided into group 1 (n = 19) with anal sphincter rupture and group 2 (n = 10) with sphincter rupture as well as pudendal neuropathy. All patients (n = 29) were examined with the Cleveland Clinic incontinence score before and median eight (2-64) months after operation. RESULTS: Median incontinence score before and after operation was 11.0 (0-22) versus 5.0 (0-16) (p = 0.002) for group 1; 18.0 (15-24) versus 15.5 (11-24) (p = 0.034) for group 2. There was also a significant difference between the groups concerning incontinence scores both preoperatively (p = 0.045) and postoperatively (p = 0.028). An improvement of continence was seen in 15 (79 %) of the patients in group 1 compared to 4 patients (40 %) in group 2. In group 1 a significant increase of median resting and squeeze pressures was seen. INTERPRETATION: Anterior sphincteroplasty improves anal continence considerably in patients with isolated sphincter rupture. However, patients with evidence of pudendal neuropathy must be informed that they should expect only a minor improvement in continence after operation. PMID- 14562776 TI - [Do short-term duration drugs for anaesthesia give postoperative advantages compared to traditional drugs?]. AB - BACKGROUND: We wanted to compare totally intravenous anaesthesia with propofol and remifentanil to mixed anaesthesia with isoflurane and fentanyl in terms of postoperative pain, nausea, length of stay and costs. MATERIAL AND METHODS: We present a prospective, non-randomised, anaesthetist-based double-blind study with a cost-utility analysis on consecutive patients undergoing laparoscopic gynaecologic day-care surgery. Premedication consisted of paracetamol and diclofenac. The mixed anaesthesia group received propofol, isoflurane, fentanyl, vecuronium, N2O and O2,the total intravenous group propofol, remifentanil, vecuronium, N2O and O2. Paracetamol and ketobemidon, metoklopramid and ondansetron were given to relieve pain and nausea, respectively. Patients were asked to report pain and nausea on a visual analogue scale after 4 and 24 hours. RESULTS: 26 mixed and 27 totally intravenous patients were comparable by age, body mass index and ASA (American Society of Anesthesiologists) scores. There was a protocol violation, as fewer totally intravenous patients received pre medication (16/26 vs. 8/27 p < 0.001). Totally intravenous patients had significantly higher consumption of ketobemidon (mean 2.35 vs. 5.43 mg/24 h p < 0.004). Within the totally intravenous group, scores for pain at 24 hours were significantly higher as compared with 4-hour scores (p = 0.012). At 24 hours, scores for nausea were significantly higher for the totally intravenous group (1.87 vs. 3.21 p = 0.014). Length of stay did not differ significantly (mean 649 vs. 729 min p > 0.2). Costs of medication were significantly higher for the totally intravenous group (mean 211 vs 285 NOK p <0.001). INTERPRETATION: The use of totally intravenous anaesthesia led to more nausea and more expenses than mixed anaesthesia in laparoscopic gynaecologic day-care surgery. No recommendations for either method can be made because of the non-randomized design. PMID- 14562778 TI - [Treatment of venous insufficiency]. PMID- 14562777 TI - [Aortic dissection--a differential diagnosis in patients with chest pain and ECG changes]. AB - BACKGROUND: The effect of thrombolytic therapy in patients with myocardial infarction is well documented. In patients presenting with chest pain it may, however, be difficult to discriminate between myocardial infarction and aortic dissection only on the basis of clinical manifestations. Moreover, patients with type A dissection may have ECG changes caused by affection of the coronary flow. MATERIAL AND METHODS: We retrospectively investigated all patients admitted to our hospital with type A dissection of the aorta over the period 1999 to March 2001. RESULTS: Fourteen patients were identified. Only two patients had normal ECG, six had ST elevation. Two patients had received antithrombotic or thrombolytic therapy. INTERPRETATION: In patients with chest pain and ST elevation, aortic dissection must be considered as a differential diagnosis before thrombolytic therapy. PMID- 14562779 TI - [Osler's disease--a risk factor for stroke]. AB - BACKGROUND: Osler's disease is an inherited disease of blood vessels that causes haemorrhage from the nasal mucosa, gastrointestinal tract, lungs or the urinary tract. Patients may also have abnormal intracerebral vessels. MATERIAL: A patient with Osler's disease and recurrent epistaxis suffered a cerebellar infarction. He had no other risk factors for stroke. An arteriovenous fistula was demonstrated in the right lung. This fistula was the probable source or aberrant pathway of an embolus to the cerebellum. The patient was treated with endovascular occlusion of the fistula to prevent further emboli to the brain. INTERPRETATION: Embolisation of thrombi from arteriovenous fistulae in the lungs is a rare cause of stroke, though important since secondary prophylaxis is very effective. Physicians should consider this possibility in the occasional patient with embolic stroke in whom no other risk factor is established. PMID- 14562781 TI - Excellent husbandry and veterinary care for laboratory animals--more important than ever. PMID- 14562780 TI - [Stenting of failing above-knee femoropopliteal percutaneous transluminal angioplasty]. AB - BACKGROUND: Percutaneous transluminal angioplasty (PTA) is increasingly used in the treatment of stenoses and short occlusions of the superficial femoral and proximal popliteal arteries. Routine treatment with arterial stents does not seem to improve the results compared to PTA without stent but the efficacy of stenting as a repair procedure of a failing PTA is not known. PATIENTS AND METHODS: In our hospital, intraluminal arterial stenting of the above knee femoropopliteal segment has been used as a repair procedure when PTA alone fails because of dissection or recoil. From 1995 to 2001, 264 above-knee femoropopliteal PTAs were performed in 203 patients. Fifteen procedures were considered failures; these cases are reported in this study. RESULTS: Arterial stents were successfully implanted in all fifteen patients. One year postoperatively, five stents had occluded, seven reconstructions were still patent. Three patients could not be followed up; they were diseased at the time of the study. INTERPRETATION: In spite of the limited patency achieved, intraluminal arterial stenting in the above-knee femoropopliteal segment can be useful as a repair procedure when PTA alone fails. PMID- 14562782 TI - [Expandable penile implants in patients with erectile dysfunction]. AB - Modern medical treatment can restore normal sexual function in the majority of men with erectile dysfunction, but some men will not obtain an erection sufficient for sexual intercourse. In some of these men, with a strong desire to have an active sexual life including intercourse, it is possible to restore the function by the use of a penile implant. We describe the indications, the surgical procedure and the results with an expandable penile implant. PMID- 14562783 TI - Recurrent Merkel cell carcinoma. PMID- 14562784 TI - Rapidly progressive tabetic neurosyphilis. PMID- 14562785 TI - Primary malignant melanoma of esophagogastric junction: a case report. PMID- 14562786 TI - Laparoscopic rectosigmod colpopoiesis as a treatment for a patient with Mayer Rokitansky-Kuster-Hauser syndrome. PMID- 14562787 TI - Prevention of mother-to-infant transmission of HIV: a new programme in China. PMID- 14562788 TI - [Immunization against tetanus in wounds]. PMID- 14562789 TI - [Incidence and prevention of occupational electrical accidents]. AB - BACKGROUND: The Norwegian Directorate for Fire and Electrical Safety receives an annual total of 50 reports of injuries caused by electricity, while the Labour Inspection Authority receives 150 reports. The underreporting is, however, considerable. In this article we estimate the Incidence of occupational electrical accidents and describe the legislation regulating the reporting of injuries from electrical accidents. MATERIAL AND METHODS: 326 of 343 electricians (95 %) returned a questionnaire on electrical accidents. RESULTS AND INTERPRETATION: We received reports that indicated an incidence of 7.6 serious accidents per 100 person-years, which equal more than 3000 electrical accidents annually in Norway. Underreporting complicates accident prevention. Doctors should more often report occupational accidents than what is common today. PMID- 14562790 TI - [Occupational injuries assessed in a physico-medical practice]. AB - BACKGROUND: In Norway over the last decade, we have seen an increase in disability claims based on occupational injury. Though much is known about the injury events, so far there have been few Norwegian studies of the relationship between work-related injuries and their somatic and functional consequences. MATERIAL AND METHODS: Over the period 1994 - 2002, 503 individuals who had filed claims for work-related musculoskeletal injuries were examined with special emphasis on the clinical consequences of the injury. The median interval between accident and examination was three years. Injury severity was scored using the Abbreviated Injury Scale (AIS). RESULTS: Falls were the most frequent type of accident; contusions, sprains and strains the most frequent injuries sustained. Many accidents were poorly documented. 16 % of the injuries were found to exceed an AIS score of 1. 30 % were working full time, while around two thirds were receiving incapacity benefits. There was no association between disability and AIS score. INTERPRETATION: In spite of the low overall level of injury severity, there was a high rate of disability subsequent to the accidents. Our results seem to indicate that injury severity is only one of several factors related to outcome after work-related injuries. PMID- 14562791 TI - [Cooperation over the boundaries of professional activities--a condition for good patient care]. PMID- 14562795 TI - A comparison of ex vivo and in vitro Sutter metacarpophalangeal prostheses, 28B: 86-91. PMID- 14562796 TI - Private, individual drug coverage in the current Medicare market. PMID- 14562797 TI - Chronic conditions and disabilities: trends and issues for private drug plans. PMID- 14562799 TI - Code of professional conduct. PMID- 14562798 TI - Adverse selection in private, stand-alone drug plans and techniques to reduce it. PMID- 14562800 TI - Office of Promoting Excellence in End-of-Life Care: Surgeon's Palliative Care Workgroup report from the field. PMID- 14562801 TI - [Meningococcal meningitis]. PMID- 14562802 TI - QSAR and the role of luck in research. PMID- 14562803 TI - Prevention of prostate cancer with finasteride. PMID- 14562804 TI - Prevention of prostate cancer with finasteride. PMID- 14562805 TI - Prevention of prostate cancer with finasteride. PMID- 14562806 TI - Prevention of prostate cancer with finasteride. PMID- 14562807 TI - Prevention of prostate cancer with finasteride. PMID- 14562808 TI - Prevention of prostate cancer with finasteride. PMID- 14562809 TI - My triathlon coach: a living example of leadership. PMID- 14562810 TI - Post-approval monitoring of animal use protocols. PMID- 14562811 TI - Monkeypox in the United States. PMID- 14562812 TI - Revisiting the assessment of IACUC effectiveness in oversight of investigator compliance. PMID- 14562813 TI - Funding opportunities for animal facility construction/renovations: preparing a successful grant application part 1. PMID- 14562814 TI - Renal arterial resistance index. PMID- 14562815 TI - Renal arterial resistance index. PMID- 14562816 TI - Helping hospice patients. PMID- 14562817 TI - Transdermal clonidine. Patching up errors. PMID- 14562818 TI - Lantus or lente insulin? The confusion builds. PMID- 14562819 TI - Treating hypernatremia. Dangerous waters. PMID- 14562820 TI - Oral vs. i.v. hydromorphone. Add a measure of caution. PMID- 14562821 TI - Using tunneled epidural catheters to treat cancer pain. PMID- 14562822 TI - Should ACE inhibition always be first-line therapy in heart failure? Lessons from the CARMEN Study. PMID- 14562823 TI - Vasodilating mechanisms of levosimendan. PMID- 14562824 TI - Compatible connections, incompatible products. PMID- 14562825 TI - A high incidence of Vitamin B12 deficiency in Israeli patients undergoing coronary angiography. PMID- 14562826 TI - Treating a fungal rash. PMID- 14562827 TI - Could my patient have deep vein thrombosis? PMID- 14562828 TI - Peripheral i.v. starts: securing and removing the catheter. PMID- 14562829 TI - Caring for a patient on pulse oximetry. PMID- 14562831 TI - Message understood. PMID- 14562830 TI - Large calcium fluxes that are not related to remodeling exist. PMID- 14562832 TI - Measure for measure. PMID- 14562834 TI - Bibliography. Current world literature. Otology and neuro-otology. PMID- 14562833 TI - Gonadotoxic effects of antitumor preparations. AB - Here we review the results of experimental and clinical observations concerning the gonadotoxic effect of antitumor chemotherapy. Previous experiments showed that antiblastic preparations with various mechanisms of cytostatic action produce damage to the sex glands that differs in the severity, stage of manifestation, and period of reparative regeneration. These differences are related to different sensitivity of epitheliocytes in the testes and structural and functional elements in the ovaries. Morphological changes in the gonads are accompanied by dysfunction of the reproductive system. We compared damages to the reproductive system and its individual components produced by various antitumor preparations. The offspring was examined after cytostatic treatment of one of the parent animals. PMID- 14562835 TI - Bibliography. Current world literature. Audiology. PMID- 14562836 TI - A nasty fright. PMID- 14562837 TI - Encapsulated sumatriptan is not bioequivalent to commercial sumatriptan. PMID- 14562839 TI - Research physical examinations: perils of brevity. PMID- 14562838 TI - Musing on Mathew et al. PMID- 14562840 TI - [Mechanical ventilation in pediatrics (III). Weaning, complications and other types of ventilation. Compications of mechanical ventilation]. AB - Mechanical ventilation can produce multiple complications. The most important acute complications are mechanical problems (respirator failure, problems with the connections and circuit, incorrect parameters or alarms), problems in the airway (disconnection, extubation, mal-positioning of the endotracheal tube, leaks, nose erosions, obstruction of the endotracheal tube due to secretions or kinking, mainstem bronchus intubation, bronchospasm, postextubation croup), pulmonary complications (ventilator-induced lung injury with barotrauma, volutrauma and biotrauma), hemodynamic complications, nosocomial infections (tracheobronchitis, pneumonia, otitis, sinusitis), failure of adjustment of the respirator to the patient, and nutritional complications. The most important chronic problems are subglottal stenosis, chronic pulmonary injury, and psychological alterations. PMID- 14562841 TI - An incredible insurance denial. PMID- 14562842 TI - [Mechanical ventilation in pediatrics (III). Weaning, complications and other types of ventilation. Noninvasive ventilation]. AB - Noninvasive ventilation (NIV), i.e. without tracheal intubation, has been reintroduced for the treatment of respiratory failure to reduce the complications of mechanical ventilation. Nowadays, NIV with positive pressure is the preferred method, applied through a mask held in place by a harness. Several masks can be used (nasal, bucconasal facial) and a variety of means can be used to keep them in place. Many respirators can be selected, ranging from those traditionally used in the intensive care unit(ICU) to specific NV respirators and conventional ICU respirators with specific software for NIV. Many respiratory modalities can be used according to the respirator (biphasic positive airway pressure [BIPAP], proportional assist ventilation, pressure support, synchronized intermittent mandatory ventilation [SIMV], etc.). NIV is mainly indicated in exacerbations of chronic respiratory failure: neuromuscular diseases, pretransplantation cystic fibrosis, and obstructive sleep apnea syndrome. It is also indicated in acute respiratory failure: pneumonia, status asthmaticus, and acute lung edema. The main contraindications are a weakened airway protection reflex(absent cough reflex) and hemodynamic instabiity. The advantages of NIV derive mainly from avoiding the complications associated with invasive ventilation. NIV also presents some disadvantages, especially the greater workload involved to ensure good patient adaptation to the respirator. The most common sequelae of NIV are skin lesions due to pressure on the nasal bridge. PMID- 14562843 TI - [Mechanical ventilation in pediatrics (III). Weaning, complications and other types of ventilation. High-frequency ventilation]. AB - In the era of lung-protective ventilation strategies, high frequency oscillatory ventilation (HFOV) has attracted renewed interest and its use has dramatically increased in neonatal and pediatric intensive care units. HFOV is able to reduce ventilator-induced lung injury by limiting the incidence of volutrauma, atelectrauma, barotrauma and biotrauma. During HFOV, adequate oxygenation and ventilation is achieved by using low tidal volumes and small pressure swings at supraphysiologic frequencies. Unlike other high-frequency ventilation modes, HFOV has an active expiration phase. HFOV constitutes a safe and successful ventilation mode for managing pediatric patients with respiratory insufficiency refractory to optimized conventional mechanical ventilation and provides better results when initiated early. However, the elective use of HFOV requires further studies to identify its benefits over conventional modes of mechanical ventilation and to support its routine use as a first line therapy. In the present article, the Respiratory Working Group of the Spanish Society Pediatric Critical Care reviews the main issues in the pediatric application of HFOV. In addition, a general practical protocol and specific management strategies, as well as the monitoring, patient care and other special features of the use of HFOV in the pediatric setting, are discussed. PMID- 14562844 TI - Lighting up cells: labelling proteins with fluorophores. AB - During the past decade, rapid improvements have been made in the tools available for labelling proteins within cells, which has increased our ability to unravel the finer details of cellular events. One significant reason for these advances has been the development of fluorescent proteins that can be incorporated into proteins by genetic fusion to produce a fluorescent label. In addition, new techniques have made it possible to label proteins with small organic fluorophores and semiconductor nanocrystals. PMID- 14562845 TI - Photobleaching and photoactivation: following protein dynamics in living cells. AB - Cell biology is being transformed by the use of fluorescent proteins as fusion tags to track protein behaviour in living cells. Here, we discuss the techniques of photobleaching and photoactivation, which can reveal the location and movement of proteins. Widespread applications of these fluorescent-based methods are revealing new aspects of protein dynamics and the biological processes that they regulate. PMID- 14562846 TI - 4D imaging to assay complex dynamics in live specimens. AB - A full understanding of cellular dynamics is often difficult to obtain from time lapse microscopy of single optical sections. New microscopes and image-processing software are now making it possible to rapidly record three-dimensional images over time. This four-dimensional imaging allows precise quantitative analysis and enhances visual exploration of data by allowing cellular structures to be interactively displayed from many angles. It has become a key tool for understanding the complex organization of biological processes in live specimens. PMID- 14562852 TI - HIV resistance to nevirapine and other non-nucleoside reverse transcriptase inhibitors. AB - Nevirapine and other members of the non-nucleoside reverse transcriptase inhibitor (NNRTI) family of anti-HIV-1 drugs are essential components of antiretroviral treatment regimens. Unfortunately, drug resistance has become an important issue with respect to all therapeutic targets in HIV-1. This paper summarizes current knowledge about the mutations in the reverse transcriptase gene of HIV-1 that are responsible for drug resistance and the mechanisms whereby drug resistance develops. PMID- 14562853 TI - Pharmacokinetic drug interactions with nevirapine. AB - Treatment of HIV infection is a multi-drug issue. Not only are there drugs for the treatment of HIV but also concomitant drugs for opportunistic infections, complications arising from the anti-retroviral therapy and other conditions related to a chronic disease. To have any understanding of drug-drug interactions in HIV treatment we need to appreciate the importance of key pharmacological areas including: 1) how each drug in a regimen is eliminated; 2) the potential for a drug to either induce or inhibit metabolic enzymes and/or transporters; 3) the therapeutic index of each drug. It is impossible to memorise all the possible drug-drug interactions in HIV, therefore understanding how drugs are metabolised/eliminated and the potential for a particular drug to modify the pharmacokinetics of another has predictive value even when substantive data are unavailable. NNRTIs interact with cytochrome P450 (CYP450) enzymes both as substrates and inducers. Because of the inductive effects caution must be exercised when using with protease inhibitors (either boosted or un-boosted with ritonavir). In this situation therapeutic drug monitoring may play a role in optimising response. There needs to be care when using many drugs with NNRTIs e.g. methadone, oral contraceptives, rifampicin, and there are some definite contraindications. By understanding pharmacological principles, it is possible to optimise use of multi-drug regimens. PMID- 14562854 TI - Defining the toxicity profile of nevirapine and other antiretroviral drugs. AB - Since there is currently no cure for HIV-1 infection, long-term remission of the disease is achieved by administering combinations of antiretroviral (ARV) agents for an indefinite period. During this time, the likelihood of developing toxicities is high, since all ARV drugs are associated with both short- and long term toxicities. The major toxicity observed with nevirapine (NVP) is rash, which is usually mild and occurs early in treatment. However, in some cases, it may be severe or potentially life threatening. Using the recommended lead-in dosing schedule in conjunction with good health care management, this adverse event can be controlled in the vast majority of patients. The development of guidelines for management of NVP-associated rash, which are easy to use, have proven to be very effective; approximately half of the patients with mild or moderate rash can continue NVP therapy under close supervision. In order to facilitate better monitoring of toxic events associated with ARV therapy, a Toxicity Evaluation Group has been set up, with the objective of standardizing reporting of drug related toxicities in all Adult AIDS Clinical Trials Group (AACTG) studies. These measures may help to identify potential toxicities better and any risk factors that predispose patients to the development of treatment-related side-effects. PMID- 14562855 TI - A comprehensive hepatic safety analysis of nevirapine in different populations of HIV infected patients. AB - All classes of antiretroviral (ARV) therapy have been associated with asymptomatic elevations of alanine aminotransferase/aspartate aminotransferase (ALT/AST) levels, and much less frequently with serious, and at times life threatening, clinical liver hepatotoxicity. The relationship between the risk of developing serious clinical liver injury and the rate and severity of elevated asymptomatic ALT/AST levels is poorly understood. Boehringer Ingelheim has recently completed the Viramune Hepatic Safety Project; its primary objective was to identify risk factors for antiretroviral-associated hepatotoxicity. Data from 1731 nevirapine-treated patients and 1912 control patients who took part in Boehringer Ingelheim-controlled clinical trials as well as 814 nevirapine-treated patients in uncontrolled trials were analyzed. Risk factors for asymptomatic ALT/AST elevations during nevirapine therapy included baseline elevations of ALT/AST levels > 2.5x upper limit of normal (RR = 4.3, p < .01) and co-infection with hepatitis B (RR = 2.3, p < .01) or hepatitis C (RR = 5.2, p < .01). An analysis of ALT/AST elevations > 5x ULN for patients stratified by baseline CD4 cell count demonstrated that men with > or = 400 CD4 cells/mm3 were at increased risk of asymptomatic transaminase elevations while taking nevirapine (RR = 1.6, p < .01). No consistent CD4 cell count cutoff could be identified in women that was associated with an increased risk of ALT/AST elevations. Analyses from five large observational cohorts (N = 8711) demonstrated no significant differences in the rate of serious hepatic events among antiretroviral regimens, including between the non-nucleoside reverse transcriptase inhibitors nevirapine and efavirenz. Use of nevirapine was not associated with a significantly increased risk of clinical hepatotoxic events, including liver failure or liver related death, compared to therapy with other antiretroviral drugs. PMID- 14562856 TI - Managing antiretroviral-associated liver disease. AB - The use of all potent drugs is associated with toxicities and antiretroviral (ARV) drugs are no exception. Antiretroviral therapy-associated hepatic toxicity is of increasing concern in the management of patients with HIV/AIDS. Liver toxicity has been reported in some HIV-infected patients being treated with drugs from all of these classes of ARV drugs: protease inhibitors (PIs), nucleoside reverse transcriptase inhibitors (NRTIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs). Although the majority of cases involve asymptomatic elevations of liver enzymes (alanine aminotransferase [ALT] and aspartate aminotransferase [AST]), severe, and, in a minority of cases, life threatening, liver disease has been reported in patients treated with ARV drugs. The exact causes of elevated plasma levels of AST and ALT are complex and, in many cases, obscure. The combination of viral hepatic disease, drugs that act adversely directly on the liver and drugs that act on other systems of the body which in turn, adversely affect the liver, can result in hepatic toxicity. Such toxicity may be inappropriately attributed solely to the direct effect of a drug. Knowledge of the possible causes of liver toxicity, and ways to avoid it, should reduce the risk of developing hepatotoxicity. The physician's task is to prevent the development of liver toxicity, e.g., by choosing appropriate therapeutic regimens and by careful management of the patient. This involves frequent monitoring of the patient, both clinically and by utilizing liver function tests on a regular basis. If signs and symptoms of liver disease do develop, prompt and expert management is essential. This review discusses the influence of a number of factors on hepatic toxicity including viral hepatitis, insulin resistance and the specific ARV drugs used in the treatment of patients with HIV/AIDS. PMID- 14562857 TI - Efficacy and durability of nevirapine in antiretroviral drug naive patients. AB - Nevirapine is a non-nucleoside reverse transcriptase inhibitor (NNRTI) that was first reported in the scientific literature in 1990. Varying doses of nevirapine (NVP) and a number of regimens containing this NNRTI have been studied in antiretroviral (ARV) naive patients. Four key studies have compared the efficacy and safety of triple drug regimens containing NVP in ARV naive, HIV-1 infected patients. The INCAS study was the first demonstration of how to use NVP in an effective and durable manner: as a component of a triple drug regimen. The COMBINE Study was a comparison of protease inhibitor (PI)-based and NVP-based triple regimens. The Atlantic Study is comparing the safety and efficacy of three triple drug regimens in ARV naive patients. In this study, treatment consists of a divergent drug regimen (PI and nucleoside reverse transcriptase inhibitors, NRTIs) targeting both HIV-1 protease and reverse transcriptase or a convergent regimen targeting reverse transcriptase alone (three NRTIs or two NRTIs plus a NNRTI). A clinical endpoint study (BI 1090) compared the efficacy and durability of multi-drug regimens in ARV naive patients with high baseline plasma HIV-1 RNA levels (pVLs) and low peripheral blood CD4+ lymphocyte counts. Data from these studies confirm that triple regimens containing NVP suppressed viral replication for up to one year, even when the ARV naive patients had low CD4+ cell counts at baseline. Nevirapine-containing regimens suppressed pVLs to < 50 copies/ mL in approximately 50% of patients in the studies discussed (Intent to Treat analyses). Data from 96 weeks of follow up in the Atlantic Study demonstrates that the regimens containing didanosine and stavudine plus indinavir or NVP were significantly more successful in suppressing pVLs to < 50 copies/mL during this period than a regimen composed of these NRTIs and lamivudine (p < or = 0.001). As with other ARV drugs, NVP should always be used as part of a fully suppressive ARV regimen. When used in this way, it is an effective ARV drug, which contributes to durable virological and immunological responses in approximately half of all treated patients. Nevirapine-containing regimens are effective in patients with advanced HIV-1 infection, i.e., low CD4+ cell counts. Data will soon be available from the 2NN Study that compares the efficacy and safety of four different regimens using NVP once daily, NVP twice daily, efavirenz once daily or a combination of NVP and efavirenz. All four arms of the study include a backbone of stavudine and lamivudine. PMID- 14562858 TI - Efficacy and durability of nevirapine in antiretroviral-experienced patients. AB - The non-nucleoside reverse transcriptase inhibitor (NNRTI) nevirapine (NVP) has been used as a component of salvage therapy for patients who have experienced virologic failure while taking nucleosides and protease inhibitors (PIs). In spite of broad cross-resistance within the NNRTI class, NVP may also play a role in salvage therapy for patients who have experienced failure while taking NNRTIs Another role for NVP in treatment-experienced patients is in so-called "PI switch" strategies, in which NVP is substituted for a PI within a virologically successful combination regimen. This paper reviews the clinical data that support the use of NVP in salvage regimens and PI switch strategies. PMID- 14562859 TI - Management of hepatitis B and C in HIV co-infected patients. AB - Morbidity and mortality from co-morbid hepatitis B (HBV) and hepatitis C (HCV) infection in HIV co-infected patients are increasing; hence, the management of HIV and HBV or HCV co-infected individuals is now one of the most challenging clinical management issues. Less than 10% of all HIV-infected patients show markers of chronic HBV infection. Hepatitis B in HIV co-infected patients is characterized by high levels of HBV replication and a high risk for cirrhosis. Treatment of HBV with lamivudine (3TC) remains the best treatment option at this time. Initial results of studies of adefovir or tenofovir, however, demonstrate good antiretroviral efficacy, even in patients with 3TC-resistant HBV. In Europe, it is estimated that approximately 30% of HIV-infected individuals are co infected with HCV. HIV accelerates HCV liver disease especially when HIV associated immune deficiency progresses. Within 10-15 years of initial HCV infection, 15-25% of patients who are co-infected with HIV develop cirrhosis compared with 2-6% of patients without HIV infection. With the introduction of pegylated interferon in combination with ribavirin, promising treatment options have become available for HIV/HCV co-infected patients leading to early virological response rates of approximately 50%. The high number of HIV/HCV and HIV/HBV co-infections, as well as the much more unfavorable course of HBV and HCV in these patients, underlines the need to establish treatment strategies for HBV and HCV in HIV co-infected individuals. PMID- 14562860 TI - Prevention of mother-to-child transmission of HIV--what next? AB - Mother-to-child transmission (MTCT) of the human immunodeficiency virus (HIV) is a global problem. HIV can be transmitted from mother-to-child at various stages of pregnancy including in utero, intrapartum and during breastfeeding. A number of interventions have, therefore, been aimed at effectively providing alternatives to breastfeeding and limiting the risk of newborn infection during delivery, by using caesarian section as the mode of delivery and administering antiretroviral (ARV) drugs prepartum and peripartum. However, these approaches are not always possible in developing countries and the use of ARV drugs, in particular nevirapine, zidovudine and zidovudine/lamivudine, have been investigated in both developing countries and developed countries. The studies have involved the administration of various ARV prophylaxis regimens to HIV infected pregnant women perinatally, either as monotherapy or in various combinations. In some studies, infants have also received ARV prophylaxis. Although studies have enrolled different populations and utilized various ARV drugs and regimens, encouraging reductions in the MTCT rates have been reported. These interventions have raised concerns regarding the development of ARV resistant HIV strains. Mutations that confer resistance to nevirapine have been detected in pregnant women who received this drug, but the emergence of these mutations was not associated with an increased risk of transmission of HIV-1 to their infants. Studies are ongoing to determine if the presence of these mutations has implications for the subsequent administration of nevirapine, either to prevent MTCT of HIV-1 or for the mother's own health. Effective interventions that can reduce MTCT of HIV are now available worldwide. However, a number of issues remain to be resolved, particularly methods to reduce the transmission of the virus during breastfeeding and to deliver effective treatment for the mothers' own HIV infection. PMID- 14562861 TI - Cardiovascular risk factors in HIV-infected patients. AB - Highly active antiretroviral therapy (HAART) commonly leads to persistent dyslipidemia and insulin resistance that appear likely to confer an increased incidence of cardiovascular disease (CVD). Both protease inhibitors (PIs) and, to a lesser extent, nucleoside analog reverse transcriptase inhibitors (NRTIs) appear to be involved, through direct metabolic effects of PIs and an indirect effect of PI and NRTI-related lipodystrophy. Several studies have found a variable relationship between CVD incidence and HAART, but these studies were not prospective and may not have been adequately powered. A variety of treatment strategies have been evaluated for dyslipidemia and insulin resistance, including lifestyle changes, drugs, and antiretroviral switching, but their relative safety, efficacy and roles are unclear. Although treatment of dyslipidemia and insulin resistance is commonly recommended, it should be remembered that such therapy is likely to be of greater benefit in those with a greater perceived CVD risk (i.e., multiple risk factors) and the lowest risk of HIV disease progression. PMID- 14562862 TI - Impact of nevirapine on lipid metabolism. AB - Abnormal blood lipid profiles may be observed both in HIV-infected individuals who are untreated and in those receiving highly active antiretroviral therapy (HAART). Besides maintaining optimal control of HIV replication and the preservation of immunity, treatment regimens ideally should have minimal or no metabolic side-effects. Nevirapine (NVP)-based HAART has beneficial effects on the lipid profile, in both treatment-naive and treatment-experienced patients, unlike protease inhibitor (PI)-based HAART. In antiretroviral (ARV)-naive patients enrolled in the Fat Redistribution and Metabolic Substudy (FRAMS) of the Atlantic Study, the NVP-containing regimen increased total cholesterol, high density lipoprotein (HDL) cholesterol concentration and particle size and apolipoprotein A1 (apo A1) levels at 24 weeks. The changes in HDL cholesterol plasma levels were demonstrated to be sustained in a subset of 98 FRAMS patients at 96 weeks. Switching from a PI-containing regimen to a PI-sparing regimen containing NVP has likewise been shown to favorably alter lipid profiles in two open label studies. In one study, one or more lipid profile parameters (total cholesterol, low density lipoprotein [LDL] cholesterol, LDL particle size, very low density lipoprotein cholesterol [VLDL1] HDL cholesterol, HDL particle size) had reverted to normal after 24 weeks in significantly more NVP-treated patients than PI-treated patients (69% versus 23%, p < .05). The 12-month results from the Barcelona PI Switch Study indicated that NVP improved lipid profiles over 12 months after PI-treated patients were switched to NVP. In conclusion, first-line NVP treatment is associated with a favorable lipoprotein profile, i.e., an increase in HDL-cholesterol and apo A1 plasma levels The lipid profile observed in patients who are switched from a PI-based regimen to a NVP-based regimen improves in a very similar fashion. These favorable lipid profiles may be of clinical benefit in reducing the risk for coronary artery disease in HIV-1 infected patients who are receiving long-term antiretroviral therapy. PMID- 14562863 TI - Mitochondrial toxicity in the era of HAART: evaluating venous lactate and peripheral blood mitochondrial DNA in HIV-infected patients taking antiretroviral therapy. AB - Nucleoside analogs can induce mitochondrial toxicity by inhibiting the human DNA polymerase gamma. This can lead to a wide range of clinical toxicities, from asymptomatic hyperlactatemia to death. Despite their technical and physiological variability, we propose that random venous lactate measurements can be useful to monitor the development of nucleoside-related mitochondrial toxicity. Recently, we have developed an assay that can measure changes in mitochondrial DNA levels in peripheral blood cells. Using this assay we have characterized changes in mitochondrial DNA (mtDNA) relative to nuclear DNA (nDNA) in peripheral blood cells of patients with symptomatic nucleoside-induced hyperlactatemia. Our results demonstrate that symptomatic hyperlactatemia was associated with markedly low mtDNA/nDNA ratios, which were on average 69% lower than HIV-uninfected controls and 45% lower than HIV-infected asymptomatic/antiretroviral naive controls. A statistically significant (p = .016) increase in mtDNA/nDNA ratio was observed following discontinuation of antiretroviral therapy. The mtDNA/nDNA ratio remained stable among selected patients who reintroduced antiretroviral therapy with stavudine (d4T)-sparing regimens. Of note, the decline in mtDNA preceded the increase in venous lactate levels. More recently we have evaluated changes in the mtDNA/nDNA ratio in relation to selected antiretroviral drug regimens in a cross-sectional study on a non-random sample of participants within the British Columbia Centre for Excellence in HIV/AIDS Drug Treatment Program. Eligible patients had continuously received saquinavir plus ritonavir with either nevirapine (n = 20), lamivudine (n = 15), d4T (n = 53) or lamivudine + d4T (n = 69), for 4 to 30 months. d4T-sparing regimens were associated with a higher median mtDNA/nDNA ratio than d4T-containing regimens (p = .016), despite the fact that study patients had received d4T-containing regimens for a shorter median time than patients taking d4T-sparing regimens (13 versus 25 months, p = .002). In summary, mtDNA levels are significantly decreased among patients who develop symptomatic, nucleoside-related hyperlactatemia, an effect reversed upon therapy discontinuation. Furthermore, mtDNA/nDNA ratios were statistically significantly lower in patients taking d4T-containing regimens than in those taking selected d4T-sparing regimens in a population setting. These results suggest that measurement of this parameter should be investigated as a potential clinical management tool. PMID- 14562864 TI - Tipranavir: a protease inhibitor from a new class with distinct antiviral activity. AB - Tipranavir (TPV) is the first of a new class of non-peptidic protease inhibitors (NPPIs). It is a sulphonamide-containing dihydropyrone, which is highly selective for the HIV protease enzyme and demonstrates potent in vitro activity against wild-type HIV-1 and HIV-2. The IC90 for TPV was 0.1 microM against clinical HIV isolates. Since CYP3A is the major cytochrome P450 isoform for the phase I metabolism of TPV, its exposure is markedly enhanced in the presence of ritonavir (RTV). In one clinical study, using the new self emulsifying drug delivery system (SEDDS) formulation of TPV, plasma concentrations in excess of 20 microM were maintained for 12 hours, allowing for twice-a-day dosing following administration of TPV 300 mg/RTV(r) 200 mg twice a day. The 20 microM target represents 10-fold the IC90 for multiple protease inhibitor (PI)-resistant strains. Both in vitro data and pharmacokinetic results indicate that TPV will be active in vivo against PI-resistant viruses, when given twice a day in combination with low dose RTV. Of 105 HIV viral isolates taken from patients who had been heavily pretreated with PI-based regimens: 90% were fully susceptible to TPV; 8% exhibited intermediate resistance; and 2% were more than 10-fold resistant. In patients who had failed at least two PI-based regimens, only 12.2% of the HIV isolates exhibited four to 10-fold reduced susceptibility to TPV after one year of treatment with a regimen containing the NPPI (Study BI1182.2). A reduction of approximately 1.5 log10 copies/mL in the plasma viral load (pVL) was observed in treatment-naive patients after 15 days of monotherapy with TPV (300 or 1200 mg twice a day) co administered with RTV (200 mg twice a day) (TPV/r) in a dose-ranging study (Study BI1182.3). The safety and efficacy of TPV (500 or 1250 mg) plus ritonavir (100 mg twice a day) plus two new nucleoside reverse transcriptase inhibitors (NRTIs) was studied in patients failing their first PI-containing regimen (Study BI1182.4). Similar decreases in pVLs (1.44-1.79 log10 copies/mL) were observed after 16 weeks of treatment with either dose of TPV/r. Two doses of TPV/r plus efavirenz (EFV) and a new NRTI have been studied in non-nucleoside reverse transcriptase inhibitor (NNRTI)-naive patients who had failed two or more PI-containing regimens (BI1182.2). Between 50% and 78.9% of patients maintained a pVL < 50 copies/mL for 48 weeks. Clinical studies have shown that TPV/r-associated adverse events are generally gastrointestinal-associated, transient and mild. A phase II study will define the optimal dose of TPV/r for highly treatment-experienced patients. The safety and efficacy of this dose of TPV/r will be evaluated in two phase III studies that will enroll more than 1300 patients worldwide. Tipranavir's robust activity against PI-resistant strains results from its molecular flexibility, which allows it to fit into the active pocket of the protease enzyme in viruses that have become resistant to other PIs. PMID- 14562865 TI - Future treatment perspectives. AB - Progress in antiretroviral therapy has resulted in dramatic achievements, such as the considerable reductions in morbidity and mortality observed in HIV-infected patients living in western countries, and new challenges, including the awareness of the long-term toxicities of antiretroviral (ARV) drugs and of viral cross resistance. Changes in treatment guidelines have occurred as a result of data obtained from clinical trials but also of a better knowledge of the pathophysiology of the disease. Recommendations on when to start therapy in asymptomatic, chronically infected individuals have shifted to a more conservative approach. The availability of new assays, particularly resistance testing, is improving the ability of physicians to select individualized treatment regimens for patients with virological failure. The availability of new ARV drugs and the evaluation of new therapeutic concepts (including treatment simplification and treatment interruptions) may impact future directions of anti HIV therapy. New drugs in development show promising antiviral activity, including activity against strains that are resistant to available agents, and this may help to circumvent cross-resistance. PMID- 14562866 TI - Research on hindsight bias: a rich past, a productive present, and a challenging future. AB - In this introduction to the present issue, we give a brief description of the phenomenon. Subsequently, we discuss the major theoretical accounts, focusing on how these are related to the papers included in the issue. PMID- 14562867 TI - SARA: a cognitive process model to simulate the anchoring effect and hindsight bias. AB - The cognitive process model "SARA" aims to explain the anchoring effect and hindsight bias by making detailed assumptions about the representation and alteration of item-specific knowledge. The model assumes that all processes, namely generating an estimate, encoding new information (i.e., the "anchor"), and reconstructing a previously generated estimate, are based on a probabilistic sampling process. Sampling probes long-term memory in order to retrieve information into working memory. Retrieval depends on the associative strength between this information and the currently active retrieval cues. Encoding the anchor may alter this associative pattern ("selective activation") or the anchor may serve as a retrieval cue, thus directing memory search ("biased reconstruction"). Both processes lead to systematically changed retrieval probabilities, thus causing the anchoring effect or hindsight bias. The model is completely formalised and implemented as a computer program. A series of simulations demonstrates the power of SARA to reproduce empirical findings and to predict new ones. PMID- 14562868 TI - Hindsight bias: how knowledge and heuristics affect our reconstruction of the past. AB - Once people know the outcome of an event, they tend to overestimate what could have been anticipated in foresight. Although typically considered to be a robust phenomenon, this hindsight bias is subject to moderating circumstances. In their meta-analysis, Christensen-Szalanski and Willham (1991) observed that the more experience people have with the task under consideration, the smaller is the resulting hindsight bias. This observation is one benchmark against which the explanatory power of process models of hindsight bias can be measured. Therefore, we used it to put the recently proposed RAFT model (Hoffrage, Hertwig, & Gigerenzer, 2000) to another test. Our findings were consistent with the "expertise effect." Specifically, we observed-using computer simulations of the RAFT model-that the more comprehensive people's knowledge is in foresight, the smaller is their hindsight bias. In addition, we made two counterintuitive observations: First, the relation between foresight knowledge and hindsight bias appears to be independent of how knowledge is processed. Second, even if foresight knowledge is false, it can reduce hindsight bias. We conclude with a discussion of the functional value of hindsight bias. PMID- 14562869 TI - Hindsight bias as a function of anchor distance and anchor plausibility. AB - This study explored the influence of anchor distance on hindsight bias and how the subjective plausibility of different anchors moderates this relation. In addition to the standard memory design used in hindsight bias research, participants were asked to indicate the range of values for possible answers to difficult almanac questions. Varying anchor distance on the basis of each participant's individual range of possible answers showed (1) that anchor plausibility decreased with increasing anchor distance following a non-linear monotone function, (2) that size of hindsight bias initially increased with increasing anchor distance but, from a certain distance, started to decrease, and (3) that hindsight bias was found to be always higher for plausible than for implausible anchors. PMID- 14562870 TI - Strength of hindsight bias as a consequence of meta-cognitions. AB - Hindsight bias is the tendency of people to falsely believe that they would have correctly predicted the outcome of an event once it is known. The present paper addresses the ongoing debate as to whether the hindsight bias is due to memory impairment or biased reconstruction. The memory impairment approach maintains that outcome information alters the memory trace of the initial judgement, whereas the biased reconstruction approach assumes that people who have forgotten their initial judgements are forced to guess and, in the presence of outcome information, are likely to use this information as an anchor. Whereas the latter approach emphasises the role of meta-cognitive considerations, meta-cognitions are not included in the memory impairment explanation. Two experiments show that the biased reconstruction approach provides a better explanation for empirical findings in hindsight bias research than does the memory impairment explanation. PMID- 14562871 TI - An inferential approach to the knew-it-all-along phenomenon. AB - Two studies tested the hypothesis that the knew-it-all-along effect may be the result of an inferential process. Specifically, that individuals use their feelings and experiences (e.g., "This question seems so familiar to me, surely I would have known the answer!") to infer their judgement. Drawing on subjective feelings such as certainty or perceptual fluency, individuals can use a provided actual value as an informational cue and draw inferences from it. Thus, the occurrence of the knew-it-all-along effect is expected to depend on the experienced feeling of confidence with a question. This feeling may indicate to an individual that he or she did know the answer; a total lack of such a feeling may suggest that he or she never would have known the answer. In the reported studies we both measured feelings of confidence (Study 1) and induced them by manipulating perceptual fluency (Study 2) to show that the knew-it-all-along effect proves to be a phenomenon of inferences based on these experienced feelings. Participants experiencing high confidence or high perceptual fluency more strongly assimilated their judgements to the provided values, than did participants experiencing low confidence or low perceptual fluency. PMID- 14562873 TI - "I couldn't have seen it coming": the impact of negative self-relevant outcomes on retrospections about foreseeability. AB - We examined a phenomenon related to hindsight bias, specifically, retrospective judgements about the foreseeability of an outcome. We predicted that negative, self-relevant outcomes would be judged as less foreseeable by the recipient of the outcome than by others, unlike either positive outcomes or outcomes that are not self-relevant. In the context of a "stock market decision-making game", the hypothetical stock selected by one of two players showed an extreme increase or decrease. As predicted, the player who received an extreme negative outcome reported that this outcome was less foreseeable than did the opponent and an observer, for whom the outcome was less self-relevant. For no other kind of outcome was there a difference between the recipient of an outcome, the opponent, and the observer. The findings have several implications, including the possibility that hindsight bias should be considered as a special case of retrospective foreseeability. PMID- 14562872 TI - Surprise, defence, or making sense: what removes hindsight bias? AB - This paper examines predictions concerning the absence of hindsight bias. Some hypothesise that because hindsight bias increases with outcome "surprisingness", only unsurprising outcomes will remove it. Others suggest the opposite-that very surprising outcomes will reduce or reverse the bias. A proposed sense-making model suggests that unexpected outcomes (i.e., initially surprising) invoke greater sensemaking, which typically produces greater hindsight bias. If the process is not successful, however, the bias may be reduced or reversed. Expected outcomes will also produce little hindsight bias, but only because they invoke relatively little sensemaking in the first place. Feelings of surprise arising from sensemaking (i.e., resultant surprise) should be inversely related to hindsight bias. Results of four experiments provide support for the model. A secondary goal was to determine the boundaries of a defensive-processing mechanism also thought to reduce hindsight bias for negative, self-relevant outcomes. Results suggest that a sense of responsibility for the outcome may be necessary for defensive processing to be activated. PMID- 14562874 TI - Hindsight bias after receiving self-relevant health risk information: a motivational perspective. AB - The phenomenon of hindsight bias was explored in the context of self-relevant health risk information. Participants in a community screening estimated their cholesterol level (foresight measure) before receiving positive or negative feedback based on their actual cholesterol level. Hindsight estimations were then assessed twice: once immediately after the feedback, and again several weeks later. While the unexpected positive feedback group showed no systematic recall bias, hindsight estimations of individuals receiving unexpectedly negative feedback showed a dynamic change over time. Immediately after the feedback, participants' recollection of their expected cholesterol level were shifted towards their actual cholesterol level (hindsight bias). In contrast, several weeks later, foresight estimations were recalled as less accurate than they had been (reversed hindsight bias). These data might reflect a change of the motivational focus from "hot affect" and fear control, which occur immediately after receiving negative feedback, to danger control, which occurs some time after the feedback, as proposed by the dual process model. PMID- 14562875 TI - Personality differences in hindsight bias. AB - Ten personality correlates of hindsight bias were tested in a study with 75 participants answering almanac-type knowledge questions. Participants showed hindsight bias when hindsight estimates were compared to foresight estimates (memory condition), when hindsight estimates were compared to foresight estimates of other participants (BS = between-subjects hypothetical condition), and when hindsight estimates were compared to foresight estimates in response to equally difficult control items (WS = within-subject hypothetical condition). The magnitude of hindsight bias in both hypothetical conditions was positively associated with the participant's field dependence and his or her tendency for favourable self-presentation (as measured by social desirability and impression management). Between-subjects hypothetical hindsight was associated with the participant's conscientiousness and need for predictability and control (as measured by a rigidity scale). In a multiple regression analysis, 39% of the variance in BS hypothetical hindsight, 24% of the variance in WS hypothetical hindsight, but no significant proportion of the variance in memory hindsight could be accounted for by personality measures. It is concluded that individual differences in hindsight bias exist and must be taken into account in a complete model of the effect. PMID- 14562876 TI - Hindsight bias in political elections. AB - Two studies on political hindsight bias were conducted on the occasions of the German parliament election in 1998 and the Nordrhein-Westfalen state parliament election in 2000. In both studies, participants predicted the percentage of votes for several political parties and recalled these predictions after the election. The observed hindsight effects were stronger than those found in any prior study on political elections (using percentage of votes as the dependent variable). We argue that the length of the retention interval between original judgement and recollection is mainly responsible for this difference. In our second study, we investigated possible artifacts in political hindsight biases using a control group design where half of the participants recalled their predictions shortly before or after the election. Hindsight bias was preserved, reinforcing the results of earlier studies with non-control-group designs. Finally, we discuss the possibility that the hindsight experience (in political judgement and in general) actually consists of three different, partly independent components. PMID- 14562877 TI - Pathogenicity of turkey coronavirus in turkeys and chickens. AB - We designed this study to compare the replication potential of turkey coronavirus (TCV) and its effect in chickens and turkeys and to study the effect of singleand combined infection of turkey poults with TCV and astrovirus. We studied the pathogenicity of TCV in experimentally inoculated turkey poults and chickens by observing the dinical signs and gross lesions. Two trials were conducted with 1 day-old and 4-wk-old specific-pathogen-free turkey poults and chickens. One-day old turkey poults developed diarrhea at 48 hr postinoculation. Poults euthanatized at 3, 5, and 7 days postinoculation had flaccid, pale, and thin walled intestines with watery contents. The 4-wk-old turkeys had no clinical signs or gross lesions. One-day-old and 4-wk-old chicks developed no clinical signs or gross lesions although the TCV was detected in gut contents of the birds throughout the experimental period (14 days). In another experiment, mean plasma D-xylose concentrations in 3-day-old turkey poults inoculated with TCV, turkey astrovirus, or a combination of both viruses were significantly lower than in the uninoculated controls. PMID- 14562879 TI - Association between cellulitis (enlarged sternal bursa) and focal ulcerative dermatitis in Ontario turkeys at the time of processing. AB - We conducted a processing plant survey to determine the prevalence of cellulitis (enlarged sternal bursa) and focal ulcerative dermatitis (FUD) in Ontario turkeys during the summer and to establish whether the two conditions were associated. A total of 11,772 birds from 24 different farms were observed at three processing plants in Ontario between June and August 2001. Each bird was examined for the presence of cellulitis and FUD. The severity and location of lesions were also noted. The overall prevalence of severe cellulitis (requiring either extensive trimming or condemnation of the bird) was 8.8% (95% confidence interval [CI] = 6.7, 10.9) and the overall prevalence for FUD was 22.6% (19.3, 25.9). The Pearson correlation coefficient between the prevalences of the two lesions was 0.56 (P = 0.0043), suggesting farms with a high prevalence of FUD in their birds also had a high prevalence of cellulitis. More specifically, birds with FUD were, on average, 10.8 times more likely to have cellulitis than birds without FUD (Mantel Haenszel summary odds ratio = 10.8; 95% CI = 9.5, 12.3). Our observations suggest FUD may predispose birds to cellulitis, in which case, preventing FUD may reduce the occurrence of cellulitis condemnations in turkeys. PMID- 14562878 TI - Characterization of a naturally occurring infection of a Mycoplasma gallisepticum house finch-like strain in turkey breeders. AB - An outbreak of Mycoplasma gallisepticum (MG) in commercial turkeys involving very mild clinical signs was difficult to confirm by routine methods. In the first part of this study (trial A), we conducted a bioassay to increase the likelihood of detecting MG. Susceptible turkeys were inoculated with sinus exudates from four different affected commercial turkey flocks. Turkeys were evaluated for clinical signs, as well as by serology and culture of tracheal swabs, at 21 and 42 days postchallenge. An MG isolate from one of the sinus exudates used for inoculation, designated K5054, was very similar to isolates from house finches when characterized by random amplified polymorphic DNA analysis as well as DNA sequence analysis of portions of the phase-variable putative adhesin protein (pvpA) gene, a lipoprotein gene, and the cytadhesin gapA/mgc1 gene. The turkeys inoculated with the K5054 sinus exudate seroconverted in the absence of severe clinical signs. There was a single reisolation of K5054 from these turkeys 42 days postchallenge. Susceptible contact turkeys were commingled with the K5054 inoculated turkeys at 49 days postchallenge. We found no evidence of transmission of MG to the contacts by culture or serology at 7, 21, or 35 days after commingling. In the second part of this study (trial B), we challenged the contacts and K5054 sinus exudate-inoculated turkeys from trial A with virulent R strain 88 days after the K5054 sinus exudate inoculation. On necropsy 10 days postchallenge, the evaluation of gross and microscopic lesions, serology, and culture showed that the turkeys previously inoculated with K5054 sinus exudate were protected against disease and reinfection. PMID- 14562880 TI - Trends in cellulitis condemnations in the Ontario chicken industry between April 1998 and April 2001. AB - We conducted a retrospective study to determine the prevalence of cellulitis condemnations in the Ontario chicken industry and the relative proportion of variation attributable to factors that vary between processors, producers, and lots and over time. The time span studied was April 1998 to April 2001. We obtained condemnation data randomly from the Chicken Farmers of Ontario and analyzed the data with a generalized mixed model. The (weighted) average prevalence of cellulitis in Ontario between April 1998 and 2001 was 0.94% (0.87%, 1.03%). The prevalence of cellulitis ranged from 0% to 14.9%, with one outlier at 30% and 95% of the data between 0 and 2.58%. The final mixed model we used to describe the variation in the prevalence of cellulitis between lots included random effect terms, the plant where the birds were processed, the producer, the quota period when the birds were processed, and the interaction term quota period by processing plant, as well as fixed effects terms, the type of inspection system and the average weight of the birds. The final model containing these variables explained approximately 78% of the total variation in the data. Our findings indicate all three random effects variables accounted for a significant amount of variation in the cellulitis data; however, the greatest source of variation was ascribed to the plants where the birds were processed. Some of the variation in cellulitis associated with processing plants was explained by the type of inspection system used by the plant, but even after controlling for this factor, there remained a relatively large amount of variation between processing plants (approximately 30%). These findings suggest there may be discrepancies in the diagnoses of the condition. Some of the variation in the prevalence of cellulitis (approximately 13%) was also attributed to the producer; however, more of the variation in the data was attributed to differences in lot-specific factors (approximately 22%). Therefore, future control efforts for cellulitis should focus on standardizing the classification of cellulitis at processing plants and identifying lot specific factors that may increase the risk of the condition. PMID- 14562881 TI - Protection and synergism by recombinant fowl pox vaccines expressing multiple genes from Marek's disease virus. AB - Recombinant fowl poxviruses (rFPVs) were constructed to express genes from serotype 1 Marek's disease virus (MDV) coding for glycoproteins B, E, I, H, and UL32 (gB1, gE, gI, gH, and UL32). An additional rFPV was constructed to contain four MDV genes (gB1, gE, gI, and UL32). These rFPVs were evaluated for their ability to protect maternal antibody-positive chickens against challenge with highly virulent MDV isolates. The protection induced by a single rFPV/gB1 (42%) confirmed our previous finding. The protection induced by rFPV/gI (43%), rFPV/gB1UL32 (46%), rFPV/gB1gEgI (72%), and rFPV/gB1gEgIUL32 (70%) contributed to additional knowledge on MDV genes involved in protective immunity. In contrast, the rFPV containing gE, gH, or UL32 did not induce significant protection compared with turkey herpesvirus (HVT). Levels of protection by rFPV/gB1 and rFPV/gl were comparable with that of HVT. Only gB1 and gI conferred synergism in rFPV containing these two genes. Protection by both rFPV/gB1gEgI (72%) and rFPV/gB1gEgIUL32(70%) against Marek's disease was significantly enhanced compared with a single gB1 or gI gene (40%). This protective synergism between gB1 and gI in rFPVs may be the basis for better protection when bivalent vaccines between serotypes 2 and 3 were used. When rFPV/gB1gIgEUL32 + HVT were used as vaccine against Md5 challenge, the protection was significantly enhanced (94%). This synergism between rFPV/gB1gIgEUL32 and HVT indicates additional genes yet to be discovered in HVT may be responsible for the enhancement. PMID- 14562882 TI - Association of Streptococcus gallolyticus strains of high and low virulence with the intestinal tract of pigeons. AB - We investigated the ability of a high virulence (STR 357) and a low virulence (STR 598) strain of Streptococcus gallolyticus to attach to the intestinal tract of pigeons. For that purpose, first of all, two groups of six pigeons were anesthetized and ligatures were placed at the beginning of duodenum, jejunum, ileum, and colon. The obtained intestinal loops of the birds of the first and second group were injected with S. gallolyticus strains STR 357 and STR 598, respectively. At 15, 30, and 60 min postinoculation, two pigeons of each group were euthanatized and the various intestinal loops were sampled for histologic, immunohistochemical, and electron microscopic examination. Both the high and low virulence strains were able to adhere to the intestinal mucosa. Indeed, all samples dearly showed numerous coccal-shaped bacteria that stained positively with S. gallolyticus antiserum and were lining up against the intestinal epithelium. Likewise, on electron microscopic examination, cocci were seen in the mucus covering the intestinal epithelium. Second, the association of S. gallyticus strains of differing virulence with the intestinal tissue was determined quantitatively. Experiments were performed as described above. The number of S. gallolyticus bacteria that adhered to the intestinal epithelium was determined by plating out 10-fold serial dilutions of the segments. No significant differences in the number of adhered bacteria were found between the strains of high and low virulence. PMID- 14562883 TI - Severity of atherosclerosis in parrots in relation to the intake of alpha linolenic acid. AB - Atherosclerosis is a common disease among parrots, but little is known about possible risk factors. Important risk factors in humans are an elevated plasma cholesterol concentration and increased platelet aggregation; high intakes of polyunsaturated fatty acids have beneficial effects. In this study, we tried to establish a relationship, if any, between dietary fatty acids and the severity of atherosclerosis in parrots. We collected dead parrots and scored the degree of atherosclerosis in the beginning of the aorta and the brachiocephalic arteries. It was not possible to assess the intake of fatty acids with food questionnaires so fatty acid composition of adipose tissue and breast muscle had to be used as an index of the dietary fatty acid composition. In all, 202 birds were collected. Gender was not related with atherosclerosis, but the degree of atherosclerosis increased with age and among the various species; African grey parrots appeared to be the most susceptible. The contents of linoleic acid in breast muscle or adipose tissue were not associated with the severity of atherosclerosis. For the relative percentage of alpha-linolenic acid in either breast muscle (P = 0.09; n = 175) or adipose tissue (P = 0.056; n = 21), a borderline significant relation with the degree of atherosclerosis was found. Parrots without atherosclerosis had significantly higher levels of alpha-linolenic acid than did the other animals. On the basis of these data, we suggest tentatively that a high dietary intake of alpha-linolenic acid protects against the development of atherosclerosis in parrots. PMID- 14562884 TI - Development of a virosome vaccine for Newcastle disease virus. AB - In an effort to protect chickens against Newcastle disease (ND), a nonreplicating virosome vaccine was produced by solubilization of Newcastle disease virus (NDV) with Triton X-100 followed by detergent removal with SM2 Bio-Beads. Biochemical analysis indicated that the NDV virosomes had similar characteristics as the parent virus and contained both the fusion and hemagglutinin-neuraminidase proteins. To target the respiratory tract, specific-pathogen-free chickens were immunized intranasally and intratracheally with the NDV virosome vaccine. This vaccine was compared with a standard NDV (LaSota) live-virus vaccine for commercial poultry. Seroconversion (> or = four fold increase in hemagglutination inhibition [HI] antibody titers) was achieved in all birds vaccinated with the virosome vaccine. Upon lethal challenge with a velogenic NDV strain (Texas GB), all birds receiving either vaccination method were protected against death. Antibody levels against NDV, as determined by enzyme-linked immunosorbent assay and HI titer, were comparable with either vaccine and increased after virus challenge. These results demonstrate the potential of virosomes as an effective tool for ND vaccination. PMID- 14562885 TI - In vitro antibiotic resistance profiles of Ornithobacterium rhinotracheale strains from Minnesota turkeys during 1996-2002. AB - Antimicrobial resistance in nearly all human and animal pathogens is on the increase. In poultry, Ornithobacterium rhinotracheale has been identified as a newly emerging respiratory bacterial pathogen that has caused significant economic losses to the poultry industry. In this study, we examined in vitro antibiotic resistance profiles of 125 isolates of O. rhinotracheale isolated from turkeys in Minnesota during 1996-2002. A majority of isolates was sensitive to clindamycin, erythromycin, spectinomycin, and ampicillin. Resistance against sulfachloropyridiazine decreased from 1996 to 2002, but an increase in resistance was seen against gentamicin, ampicillin, trimethoprim sulfa, and tetracycline. The annual trend slopes for these antibiotics were 7.36%, 3.02%, 2.43%, and 1.95%, respectively. The resistance against penicillin remained constant from year to year with a trend slope of only 0.54% per year. These results emphasize the need for continued monitoring of O. rhinotracheale isolates for antibiotic resistance and establishment of baseline resistance pattern data for this organism. These data can then be used to design and evaluate local epidemiological interventions. PMID- 14562886 TI - Establishment of persistent avian infectious bronchitis virus infection in antibody-free and antibody-positive chickens. AB - Avian infectious bronchitis virus (IBV) causes a highly contagious and economically significant disease in chickens. Establishment of a carrier state in IBV infection and the potential for the persistent virus to undergo mutations and recombination in chicken tissues have important consequences for disease management. Nevertheless, whether chickens can maintain persistent IBV infection in the absence of reinfection from exogenous sources or the presence of antibody in the host can modulate virus persistence remains unclear. Indeed, whether or not IBV genome can undergo genetic changes during in vivo infection has not been demonstrated experimentally. In the present study, IBV shedding and tissue persistence were monitored in individual chickens maintained under strict isolation that precluded reinfection from exogenous sources. In the first of two experiments, intranasal exposure of 6-wk-old antibody-free chickens to IBV vaccine virus resulted in intermittent shedding of the virus from both trachea and cloaca of individual birds for up to 63 days. Also, the virus was recovered from the internal organs (spleen, gonad, kidney, lung, cecal tonsil, and cloacal bursa) of six of eight birds killed at various intervals between 27 and 163 days postinoculation (DPI). In the second experiment, IBV exposure of 1-day-old maternal antibody-positive chicks led to periodic virus shedding from the trachea and cloaca in all chickens until 77 days; however, internal organs (lungs and kidneys) of only one of seven birds (killed at 175 DPI) were virus positive, suggesting that presence of antibody at the time of infection protects internal organs from IBV infection. When the lung and kidney isolates of IBV from the latter experiment were compared with the parent-vaccine virus, no changes in their antigenicity, tissue tropism, or the nucleotide sequence of the S1 glycoprotein gene were observed. These findings indicate that, unlike the mammalian coronaviruses, propensity for frequent genetic change may not be inherent in the IBV genome. PMID- 14562887 TI - Clinical and pathologic features of West Nile virus infection in native North American owls (Family strigidae). AB - Since the initial report of West Nile virus in the northeastern United States in 1999, the virus has spread rapidly westward and southward across the country. In the summer of 2002, several midwestern states reported increased cases of neurologic disease and mortality associated with West Nile virus infection in various native North American owl species. This report summarizes the clinical and pathologic findings for 13 captive and free-ranging owls. Affected species were all in the family Strigidae and included seven snowy owls (Nyctea scandiaca), four great-horned owls (Bubo virginianus), a barred owl (Strix varia), and a short-eared owl (Asio flammeus). Neurologic signs identified included head tilt, uncoordinated flight, paralysis, tremors, and seizures. Owls that died were screened for flaviviral proteins by immunohistochemical staining of formalin-fixed tissues, followed by specific polymerase chain reaction assay to confirm West Nile virus with fresh tissues when available. Microscopic lesions were widespread, involving brain, heart, liver, kidney, and spleen, and were typically nonsuppurative with infiltration by predominantly lymphocytes and plasma cells. Lesions in owls were much more severe than those previously reported in corvids such as crows, which are considered highly susceptible to infection and are routinely used as sentinel species for monitoring for the presence and spread of West Nile virus. This report is the first detailed description of the pathology of West Nile virus infection in Strigiformes and indicates that this bird family is susceptible to natural infection with West Nile virus. PMID- 14562888 TI - Interactions between the membranes of turkey cells and Mycoplasma meleagridis. AB - We used Myoplsma meleagridis (MM) to infect the RP-9 cells and the eggshell membranes and scanning electron microscopy (SEM) and confocal microscopy to study the interactions between the organisms and the cell surfaces. The surface of the RP-9 cells contained numerous projections. After 24 hr of infection with MM, those projections were either lost or aggregated to the side; MM-like particles could be seen on the surface of the cells, and surface fluorescence could be detected by confocal microscopy. On the surface of MM-infected shell membranes were necrotic fibrous tissues and cells detected by SEM and an intense surface fluorescence detected by confocal microscopy. These results indicate that MM infection of the cell surface can result in cellular damage. PMID- 14562889 TI - The effects of avian leukosis virus subgroup J on broiler chicken performance and response to vaccination. AB - The effects of avian leukosis virus subgroup J (ALV-J) infection on meat-type chickens reared in a simulated commercial setting were evaluated. Each of three ALV-J isolates was evaluated with both simulated horizontal transmission (SHT) and simulated vertical transmission (SVT). Mortality, morbidity, disease condemnations, and feed conversions were increased and body weights at processing were decreased in ALV-J infected birds as compared to sham inoculated hatch mates. The adverse effects of ALV-J infection were more severe in birds exposed by SVT than in birds exposed by SHT. At 8 weeks of age response to vaccination for infectious bronchitis virus and Newcastle disease virus or prior exposure to a pathogenic reovirus was assessed in the ALV-J and sham inoculated broiler chickens by challenge studies. Although not statistically significant, an overall trend of decreased protection to challenge after vaccination, or prior exposure, was observed in the ALV-J inoculates as compared to sham inoculated hatch mates. Differences in vaccine response were most evident in groups inoculated with ALV-J by the SVT route. PMID- 14562890 TI - Attenuation, safety, and efficacy of an infectious bronchitis virus GA98 serotype vaccine. AB - In 1998, novel strains of infectious bronchitis virus (IBV) were identified in chickens from the southeastern United States and classified as a new serotype designated Georgia 98 (GA98). Because of the widespread nature of the GA98 virus in the southeastern United States and the lack of adequate protection with the DE072 vaccine, we developed a specific vaccine for the GA98 serotype. The GA98/0470/98 isolate of IBV was passaged in embryonating chicken eggs 70 times, and attenuation of the virus was determined in specific-pathogen-free chicks. Pass 70 of the GA98/0470/98 strain of IBV when given at 1 day of age by coarse spray and at 14 days of age in the drinking water at 1 x 10(4.5) 50% embryo infectious dose/bird protected against the homologous GA98 challenge as well as provided good protection against the DE072-type virus. In addition, the vaccine was shown to be adequately attenuated and safe at a 10x dosage. PMID- 14562891 TI - Effect of prior serial in vivo passage on the frequency of Salmonella enteritidis contamination in eggs from experimentally infected laying hens. AB - Experimental infection models are valuable tools for understanding and preventing the deposition of Salmonella enteritidis inside eggs. Oral inoculation is believed to closely simulate naturally occurring S. enteritidis infections of chickens, but oral infection studies have often generated relatively low frequencies of egg contamination. The present study assessed whether repeated in vivo passage of an S. enteritidis strain could affect its ability to cause egg contamination in experimentally infected hens. The incidence of egg contamination was determined in groups of hens inoculated orally with either a phage type 13a S. enteritidis strain or derivatives of this parent strain that were obtained by three successive rounds of passage and reisolation from tissues of infected hens. Passaged S. enteritidis isolates recovered from ovaries and oviducts induced a significantly higher incidence of egg contamination (16.97%) than was attributed to the parent strain (8.27%). However, passaged S. enteritidis isolates recovered from livers and spleens were not associated with a significantly increased frequency of deposition in eggs. By either inducing or selecting for the expression of relevant microbial properties, passage of S. enteritidis through reproductive tissues of chickens may be useful for improving the efficiency at which experimental infection models produce egg contamination. PMID- 14562892 TI - Molecular variability of house finch Mycoplasma gallisepticum isolates as revealed by sequencing and restriction fragment length polymorphism analysis of the pvpA gene. AB - Mycoplasma gallisepticum, a major pathogen of chickens and turkeys, has caused significant declines in house finch (Carpodacus mexicanus) populations in the eastern United States since it was first observed in this species in 1994. There is evidence that M. gallisepticum infection is now endemic among eastern house finches, although disease prevalence has declined, suggesting an evolving host parasite relationship. Studies based on randomly amplified polymorphic DNA (RAPD) have documented the presence of a single, unique RAPD profile in house finch M. gallisepticum isolates, suggesting a single point source of origin, which agrees with the known epidemiologic observations. In the present study, we evaluated the molecular variability of 55 house finch isolates as well as 11 chicken and turkey isolates including reference strains of M. gallisepticum. Molecular variability was evaluated by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis and nucleotide sequencing of the pvpA gene, which encodes for the putative cytadhesin protein PvpA. Three different RFLP groups and 16 genotypes were evident from the 55 house finch isolates evaluated. Sequence analysis of pvpA gene PCR products showed that although most house finch M. gallisepticum isolates clustered more closely to each other, others clustered more closely to either turkey or chicken field isolates. These findings suggest that house finch isolates are more polymorphic than previously recognized by RAPD studies. This feature may allow us to learn more about the molecular evolution and epidemiology of this emerging disease host-parasite relationship. PMID- 14562893 TI - Pasteurella multocida-associated sinusitis in khaki Campbell ducks (Anas platyrhynchos). AB - Pasteurella multocida group B, serotype 3, was isolated from sinusitis-affected khaki Campbell ducks. To study the role of P. multocida in sinusitis, commercial khaki Campbell ducks were experimentally infected with P. multocida alone or combined with Escherichia coli. In Expt. 1, experimental ducks were infected with P. multocida intranasally or ocularly. A comparison was done by intranasal inoculation with pooled nasal discharge from the affected ducks or phosphate buffered saline. The ducks intranasally inoculated with the nasal discharge or P. multocida showed sinusitis. In Expt. 2, E. coli alone or a combination of P. multocida and E. coli was intranasally inoculated into experimental ducks. The ducks intranasally inoculated with the combination of P. multocida and E. coli had sinusitis, the same as found in the field but less severe than that of the field cases. Pasteurella multocida was already present in litter/floor of duck farms. We concluded that P. multocida played a role in induction of sinusitis. However, the sinusitis in ducks may be initiated by poor management, especially in the brooding period of ducks. PMID- 14562894 TI - Use of a live attenuated Salmonella typhimurium vaccine to protect hens against Salmonella enteritidis infection while undergoing molt. AB - Previous studies demonstrated that Salmonella enteritidis infections in hens undergoing molt via feed withdrawal were more severe than in full fed hens. We conducted two trials to determine if immunizing specific-pathogen-free, Salmonella-culture-negative hens via aerosol exposure to MeganVacl, a commercially available attenuated Salmonella typhimurium vaccine, would reduce transmission of S. enteritidis from infected hens to uninfected but contact exposed hens during a molt. In trial 1, one group of hens received two aerosol doses of vaccine 2 wk apart whereas a second group of hens remained nonvaccinated. In trial 2, the vaccinated group received only one dose of vaccine. Two weeks after the final immunization, feed was removed from all the hens, and on day 4, the center hen in rows of 11 hens received a dose of 3 x 10(5) (trial 1) or 1.3 x 10(6) (trial 2). Transmission to the unchallenged hens was followed 3, 10, 17, and 24 days later. Vaccination reduced the horizontal spread of S. enteritidis in vaccinated hens compared with their nonvaccinated counterparts, with vaccinated hens shedding significantly less S. enteritidis on day 10 postchallenge in trial 1 and on days 3, 10, 17, and 24 in trial 2. Recovery of S. enteritidis from ovaries was significantly reduced in the vaccinated hens in trial 1 and from livers/spleens, ovaries, and cecum in trial 2. These studies indicate that immunization of hens with a live S. typhimurium vaccine could help reduce S. enteritidis problems during a molt situation. PMID- 14562895 TI - Detection of chicken anemia virus DNA in embryonal tissues and eggshell membranes. AB - Chicken infectious anemia virus (CIAV) is a ubiquitous and highly resistant virus of chickens that causes anemia and death in chicks less than 3 wk of age and immunosuppression in chickens older than 3 wk of age. The production of specific pathogen-free eggs free of CIAV is essential for research and vaccine production. Currently, flocks are screened for CIAV by antibody tests to ensure freedom from CIAV infection. Recent evidence, however, indicates that chickens may carry and vertically transmit CIAV DNA independently of their antibody status. In this study, we tested embryos and eggshell membrane residues by nested polymerase chain reaction (PCR) as a sensitive method of detecting CIAV DNA. CIAV DNA could be detected in the blastodisks and semen obtained from antibody-positive and negative chickens. Examination of different tissues between 18 and 20 days of incubation indicated that many but not all organs of individual embryos were positive. The lymphoid organs and gonads had the highest incidence of CIAV DNA, which was significantly different (P < 0.05) from the incidence in the liver. Eggshell membrane samples from embryos or newly hatched chicks were an excellent noninvasive source for the detection of CIAV DNA, identifying significantly more positive embryos than did pooled lymphoid organs. The use of dexamethasone injections as a method to improve the detection of carrier birds did not result in an increase of vertical transmission or cause seroconversion in the treated hens. A combination of testing eggshell membrane residues at hatch and periodic testing of blood DNA by nested PCR can be used to identify chickens carrying CIAV DNA and may be used to eradicate carrier birds. PMID- 14562896 TI - Comparison of the intravenous chicken challenge method with the embryo lethality assay for studies in avian colibacillosis. AB - In previous studies, the embryo lethality assay (ELA) was able to discriminate between virulent and avirulent avian Escherichia coli isolates and to predict percent mortality of the embryos resulting from an isolate based on three traits. The abilities to resist host complement, presence of Colicin V activity, and presence of the increased serum survival gene cluster (iss), were used together in a logistic regression analysis to predict the percentage of embryo deaths resulting from each of 20 avian E. coli isolates used in the ELA. In the present study, the same 20 isolates are used in an intravenous chicken challenge model in an effort to determine whether the ELA could be used to replace chicken challenge studies. Correlations between the mortality and a combination of mortality and morbidity (the survivors at trial termination with lesions suggestive of colibacillosis) and the previous ELA results and with selected isolate traits were performed. Additionally, resulting body weights in surviving chickens were compared between groups. The highest positive correlations were observed between the ELA and the combined mortality/morbidity of the intravenous challenge (r = 0.861, P < 0.0001 for the first ELA challenge, and r = 0.830, P < 0.0001 for the second ELA challenge). The IV challenge combined mortality/morbidity results had the highest correlation coefficients with the presence of iss (r = 0.864, P < 0.0001) and the expression of ColV (r = 0.878, P < 0.0001). The presence of tsh was slightly correlated with mortality (r = 0.465, P = .0389) but demonstrated a higher correlation with the combined mortality and morbidity of the IV challenge (r = 0.558, P = 0.0106). Even though the ELA results in a higher number of nonspecific deaths, the two challenge methods exhibit similar results and a high correlation with each other. Interestingly, some of the isolates showed differences in their ability to cause mortality between the ELA and the IV challenge model. Furthermore, some isolates reflected significant differences in body weights of surviving birds at IV trial termination. PMID- 14562897 TI - Infectious bursal disease virus and proventriculitis in broiler chickens. AB - Acute necrotic proventriculitis is a naturally occurring disease of broiler chickens that causes proventricular rupture during routine evisceration. Although infectious bursal disease virus (IBDV) has been implicated, it has not been proven to be a direct cause of this disease. To further study the role of IBDV in proventriculitis, proventriculi and bursas were collected during both acute and chronic phases of naturally occurring proventriculitis and from chickens experimentally infected with seven different [BDV strains. All tissues were examined for IBDV by light microscopy, immunohistochemistry (IHC), and real time reverse transcriptase(RT)-polymerase chain reaction (PCR) and for apoptosis by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling method (TUNEL). Tissues from naturally occurring proventriculitis had bursal and proventricular lesions. Two out of four bursas had no IHC stainable IBDV antigen or RT-PCR detectable IBDV sequences. No proventriculus had IBDV detectable by any of these methods. Bursas from chickens experimentally infected with IBDV had microscopically evident lesions, IBDV was detectable by IHC and RT-PCR, and strong IHC staining for apoptosis was present. Proventriculi from these experimentally exposed chickens had no lesions, low levels of IBDV detectable by IHC or RT-PCR, and very little IHC-stainable apoptosis. We conclude that naturally occurring proventriculitis can occur in the absence of IBDV and that the IBDV strains tested do not directly produce proventriculitis or induce increased proventricular apoptosis. PMID- 14562899 TI - Seroprevalence of avian pneumovirus in Minnesota turkeys. AB - Avian pneumovirus (APV) causes respiratory tract infection in turkeys and was first seen in the United States in Colorado in late 1996. In early 1997, the disease was recognized in Minnesota and caused estimated losses of up to 15 million dollars per year. This virus has not been reported in the other turkey producing states. We here report the seroprevalence of APV in Minnesota from August 1998 to July 2002. The average rate of seroprevalence has been 36.3% (range = 14.2%-64.8%). A seasonal bias was observed, with peak incidences in the fall and spring. A higher rate of seropositivity was observed in counties with the highest concentration of turkeys. PMID- 14562898 TI - Evaluation of two mycotoxin binders to reduce toxicity of broiler diets containing ochratoxin A and T-2 toxin contaminated grain. AB - In order to assess ochratoxin A (OA) and T-2 toxin (T-2) binding ability of two commercial sorbents, both in vitro and in vivo trials with broilers were performed. Crude OA and T-2 extracts from contaminated grain were used to assess in vitro binding ability of two sorbents (Zeotek [Zk] and Mycofix [Mx]), by quantifying free mycotoxin through an enzyme-linked immunosorbent assay (ELISA) test. For in vivo trial, a 3 x 2 x 2 factorial arrangement was used for this experiment, being the factors: adsorbents (none, Zk, and Mx), OA (0 and 567 parts per billion [ppb]) and T-2 (0 and 927 ppb). OA and T-2 contaminated wheat and corn, respectively, were added to sorghum-soybean meal diets to meet 567 ppb of OA and 927 ppb of T-2. Mycotoxins were fed alone or combined in treatments. After 21 days, blood chemistry, gross, and histological evaluations were performed. Relative weights of liver, kidney, and bursa of Fabricius were obtained. Zk had the highest OA and T-2 in vitro binding ability (100% and 8.67%, respectively). Chickens fed OA with or without sorbents had a lower body weight and feed intake reduction. However, those birds fed T-2 were partly protected by a sorbent. Birds fed both toxins showed toxic additive effects, and no protection of any adsorbent was observed. A significant reduction in plasma proteins, albumin, and globulins was a characteristic observed in all birds fed diets with OA both with or without adsorbents. Uric acid level in blood was increased in all chickens fed OA contaminated diets. Histological findings observed in birds fed OA-contaminated diets were necrosis of kidney tubular cells, swollen and necrotic hepatocytes, bile ducts hyperplasia, and increased diameter of proventriculus glands. In birds that received T-2 alone, only the liver, with the same kind of lesions, was affected. According to these results, it can be concluded that there is not a relation between in vitro and in vivo trials. OA toxic effects could not be counteracted by any sorbent. T-2 toxicity could be partially counteracted by an adsorbent used in this research. PMID- 14562900 TI - Incidence and tracking of Clostridium perfringens through an integrated broiler chicken operation. AB - Clostridium perfringens has been shown to be widespread in the broiler chicken hatchery, grow-out, and processing operations. In a previous study, ribotypes of certain strains of C. perfringens isolated from processed chicken carcasses were shown to match ribotypes isolated from paper pad lining trays used to transport commercial chicks from the hatchery to the grow-out facility on the farm. These results suggest that C. perfringens contaminating the processed product could originate from facilities in the integrated poultry operation prior to grow out. In this study, samples were collected from the breeder farm, hatchery, previous grow-out flock, during grow out and after processing. In the first trial, C. perfringens was recovered from the breeder farms, the hatchery, previous grow-out flock, grow-out flock at 3 weeks of age, grow-out flock at 5 weeks of age, from processed carcasses, and from the breeder farm after processing in 4%, 30%, 4%, 0%, 2% and 16%, and 4% of the samples, respectively. In the second trial, the incidence of C. perfringens in samples collected from breeder farms, the hatchery, previous grow-out flock, grow-out flock at 3 weeks of age, grow-out flock at 5 weeks of age, and fromprocessed carcasses was 38%, 30%, 32%, 8%, 4%, and 8%, respectively. The genetic relatedness of the isolated strains as determined by ribotyping suggests that C. perfringens may be transmitted between facilities within the integrated broiler chicken operation. PMID- 14562901 TI - Real-time polymerase chain reaction for Mycoplasma gallisepticum in chicken trachea. AB - In this work, we describe a rapid detection procedure for Mycoplasma gallisepticum from chicken tracheal swabs by real-time polymerase chain reaction (PCR) by LightCycler system, where we were able to monitor the amplification of the newly synthesized M. gallisepticum-specific PCR product as a proportionally increasing fluorescent signal by using the double-stranded DNA binding dye SYBR Green I and have identified M. gallisepticum-specific PCR products by DNA melting curve analysis by plotting the first negative derivative (-d[F1]dT) of fluorescence over temperature. Detection limits of the PCR were found to be 3 and 3000 colony-forming units ml(-1) with pure culture of M. gallisepticum and artificially spiked samples, respectively. Out of 96 tracheal swabs, 68 were taken from live chickens and 28 were taken by scraping the mucosal surface of the trachea (SMST) of necropsied chickens. All of the 18 PCR-positive results were from the swabs taken by the SMST method, whereas all of the samples taken from live chickens were negative. Thus, the PCR with the SMST method had a sensitivity and a specificity of 64.2% (18 of 28 chickens) and 100%, respectively. The total time required for template preparation from tracheal swab samples and real-time PCR was approximately 65 min. These results indicate that real-time PCR with the LightCycler technology is a rapid and sensitive test to identify M. gallisepticum infected flocks if a proper sampling is applied. PMID- 14562902 TI - Detection of infectious bronchitis virus by real-time reverse transcriptase polymerase chain reaction and identification of a quasispecies in the Beaudette strain. AB - In this report, we describe a real-time reverse transcriptase-polymerase chain reaction (RRT-PCR) diagnostic test for infectious bronchitis virus (IBV) with the use of fluorescence resonance energy transfer (FRET) technology. Two primers that amplify a 383-base pair product between nucleotide positions 703 and 1086 relative to the start codon for the S1 gene of the Massachusetts 41 virus were designed and used to amplify the Beaudette, Massachusetts 41, Florida 18288, Connecticut, Iowa 97, Arkansas DPI, CA/NE95/99, DE/072/ 92, and GA/0470/98 strains of IBV. The primers were specific and did not amplify New Castle disease virus, Mycoplasma spp., or infectious laryngotracheitis virus. For RRT-PCR by FRET, an anchor probe conjugated to fluorescein and a detection probe conjugated to a red fluorophore were designed to anneal to a hypervariable region within the 383-base pair product. The level of sensitivity was 1 x 10(4) RNA molecules used as starting template. After amplification, a melting curve analysis was conducted to specifically identify IBV types. Because of sequence differences in the annealing position of the detection probe, the Arkansas, Connecticut, Beaudette, and Massachusetts 41 strains could be differentiated. No fluorescence was observed for the DE/072/ 92 and GA/0470/98 viruses with the anchor and detection probes. When the Beaudette strain was examined, two melting peaks were observed at 44 C and 51 C, indicating a quasispecies in that laboratory strain of IBV. Routine typing of vaccine strains of IBV was possible with this technology, but high standard deviations associated with the melting curve analysis of the FRET probes described herein made it difficult to use this test reliably for routine typing of IBV field isolates. PMID- 14562903 TI - Characterization of infectious bronchitis virus isolates by slot blot hybridization. AB - We used slot blot hybridization of the hypervariable regions of the S1 subunit of spike peplomer gene to identify and characterize infectious bronchitis virus (IBV) strains. Template DNA was created from six reference strain IBVs of different serotypes and immobilized on a nitrocellulose membrane. We synthesized digoxigenin-labeled probes from reference and unknown field viruses and hybridized them to template DNA. All reference strains could be distinguished and isolates identified by serotype if they were at least 95% identical to a reference strain. This slot blot hybridization procedure was specific and reproducible, and strain typing was consistent with the S1 sequencing of the IBV genome. This study thus provides a simple and rapid method for typing of IBV. PMID- 14562904 TI - Prevalence of antibodies against Ornithobacterium rhinotracheale in broilers and breeders in Southern Brazil. AB - In this investigation, we determined the prevalence of the Ornithobacerium rhinotracheale (ORT) infection in broilers and broiler breeders in southern Brazil. We also correlated the presence of antibodies in broilers with performance. Sera from 1550 broilers from 50 flocks were collected during the slaughter time in nine companies with federal veterinary inspection of the state of Rio Grande do Sul. Sera from 480 meat-type breeders of 40 flocks from 14 companies in southern Brazil were also analyzed by enzyme-linked immunosorbent assay, and the prevalence of antibodies was determined. The prevalence of ORT antibodies in broiler flocks was 63.83%, but in each individual flock only 6.52% of the birds were positive. The prevalence in broiler breeder flocks was 100.00%, and in each individual flock 94.62% of the birds were positive. There was a positive correlation between the presence of antibodies to ORT and decreased body weight in broilers. There was no significant correlation between presence of antibodies to ORT and age, lineage, efficiency index, feed conversion, and mortality. There was a positive correlation between the presence of respiratory signs and antibodies to ORT, although the reverse correlation was not significant. These results confirm that ORT is present and widespread in broilers and broiler breeders in southern Brazil. PMID- 14562906 TI - Induction of a mycoplasma gallisepticum pMGA gene in the chicken tracheal ring organ culture model. AB - Genetic and molecular methods to investigate the pathogenesis of the poultry respiratory pathogen Mycoplasma gallisepticum are quite limited. Therefore, the objective of this study was to design and evaluate a functional genomics approach to identify M. gallisepticum genes involved in colonization of the poultry respiratory tract. To serve as a transcriptional reporter, a promoterless lacZ gene from Escherichia coli was cloned into the Tn4001 transposon. The transposon was used to randomly mutagenize the chromosome of the M. gallisepticum S6 strain, and a bank of 1386 transposon mutants containing lacZ fusions to mycoplasma chromosomal DNA was assembled. Each mycoplasma clone containing the lacZ reporter was independently screened in the chicken tracheal ring organ culture (TROC) model system for increased production of beta-galactosidase. A twofold or greater increase in beta-galactosidase was consistently observed for eight mutants. In one of the mutants, the transposon was inserted in a pMGA gene encoding a cell surface adhesin involved in hemagglutination. Therefore, these data indicate that screening of a M. gallisepticum transposon reporter bank with a chicken TROC model is useful for the identification of genes induced during poultry colonization and virulence. PMID- 14562905 TI - Real-time reverse transcriptase-polymerase chain reaction detection and analysis of nucleotide sequences coding for a neutralizing epitope on infectious bursal disease viruses. AB - We used real-time reverse transcriptase (RT)-polymerase chain reaction (PCR) to detect infectious bursal disease virus (IBDV) strains. The LightCycler (Roche) and hybridization probe system (Roche, Molecular Biochemicals) were used. A mutation probe labeled with fluorescein and an anchor probe labeled with Red-640 dye were prepared for each of the STC, Del E, D78, and Bursine 2 viral sequences. The mutation probes were designed to hybridize to nucleotides that encode the hydrophilic B region of VP2 for each virus. The anchor probes were designed to a relatively conserved region immediately downstream from the mutation probes. When hybridized to the RT-PCR product, a mutation and anchor probe pair produced fluorescence resonance energy transfer that was detected by the LightCycler instrument. Because they were designed to have a lower melting temperature (Tm), the mutation probes dissociated from the template before the anchor probes. The Tm values of the four mutation probes for each of their homologous viruses (exact sequence match) were STC, 69.3 +/- 1.2 C; D78, 67.8 +/- 0.9 C; Del E, 65.5 +/- 0.6 C; and Bursine 2, 71.7 +/- 0.4 C. These values were compared with the Tm values observed for a particular probe and heterologous virus. If the Tm values observed for heterologous viruses were within two standard deviations of the Tm for the probe and its homologous virus, the nucleotide sequences of the viruses were considered to be similar. If they were below two standard deviations, they were considered to have one or more nucleotide mutations. The results indicated that the STC and Variant Vax BD viruses have similar genetic sequences at the hydrophilic B region. Likewise, Bursine 2, Bursine, Bursine+, BioBurs, BioBurs W, BioBurs AB, and IBDV Blen have similar nucleotide sequences in this region. The Tm values obtained for the D78 and Del E mutation probes with heterologous viruses indicated that none of the viruses tested had nucleotide sequences that matched these probes. Because the mutation probes were designed to bind to a region that encodes a neutralizing epitope, viruses with similar sequences were expected to have antigenically similar epitopes. PMID- 14562907 TI - Absence of humoral response in flamingos and red-tailed hawks to experimental vaccination with a killed West Nile virus vaccine. AB - Sixteen Chilean flamingos, Phoenicopterus chiles, and 10 red-tailed hawks, Buteo jamacensis, were vaccinated in the pectoral muscle with 0.2 ml of a commercially produced killed West Nile virus vaccine intended for use in horses. Half the birds of each species received a booster vaccination 3 weeks after the first injection. Three weeks after the booster vaccination, none of 13 birds surveyed had detectable antibody to West Nile virus. PMID- 14562908 TI - Failure of viable nonculturable Campylobacter jejuni to colonize the cecum of newly hatched leghorn chicks. AB - Campylobacter jejuni cells entered the viable but nonculturable (VBNC) state upon suspension in sterile water. Cell viability was determined with tetrazolium violet. VBNC cells suspended in water for 7, 10, or 14 days were given, by gavage, to day-of-hatch leghorn chickens. The ceca of control and challenged birds were examined for the presence of campylobacteria by conventional microbiological methods at 1 wk and 2 wk after challenge inoculation and by polymerase chain reaction methods at 1 wk after challenge. We did not find culturable Campylobacter cells in the ceca. Neither was Campylobacter DNA found in cecal samples. Therefore, VBNC cells did not revert to the culturable colonizing form, nor did VBNC cells persist within the cecal environment. PMID- 14562909 TI - Beneficial effect of beta-mannanase feed enzyme on performance of chicks challenged with Eimerla sp. and Clostridium perfringens. AB - Two experiments were conducted to determine the effect of a beta-mannanase feed enzyme on the performance of broiler chicks subject to a necrotic enteritis disease challenge model involving oral inoculation of Eimeria sp. and Clostridium pefringens. Beta-mannanase is known to improve productive performance when added to poultry and swine diets. In both experiments, disease challenge in the absence of feed additives demonstrated significant reductions in performance as measured by weight gain, feed conversion, and the incidence of coccidial lesion scores. Significant mortality was also observed in challenged groups in Experiment 1. The disease challenge model was therefore judged as highly effective. Additions of a commonly used antibiotic, bacitracin methylene disalicilate (BMD), and coccidiostat, salinomycin, were highly effective in partially counteracting negative effects of the disease challenge. In both experiments, addition of beta mannanase significantly improved performance and reduced lesion scores in disease challenged groups. The degree of improvement was somewhat less than that afforded by a combination of BMD and salinomycin in Experiment 1 but was not different from that afforded by BMD alone in Experiment 2. We conclude that the beta mannanase enzyme can play a role in circumstances where the use of antibiotics is not desired. PMID- 14562910 TI - Avian pneumovirus and its survival in poultry litter. AB - The survival of avian pneumovirus (APV) in turkey litter was studied at different temperature (room temperature, [approximately 22-25 C], 8 C, and -12 C) conditions. Built-up turkey litter from a turkey breeder farm known to be free of APV was obtained and was divided into two portions. One portion was sterilized by autoclaving and the other portion was kept nonautoclaved. Both samples were inoculated with a Vero cell-propagated Minnesota isolate of APV subtype C (APV/MN2A) with a titer of 10(5) 50% tissue culture infective dose at 1% level. These samples were then stored at three different temperatures: -12 C, 8 C, and room temperature (20-25 C). The samples were tested for the presence of viral RNA by reverse transcriptase-polymerase chain reaction and for the presence of live virus by virus isolation in Vero cells at the intervals of 1, 2, 3, 7, 14, 30, 60, and 90 days. Our studies revealed the presence of APV RNA even after 90 days in the autoclaved litter samples kept at -12 C and at 8 C. The virus was isolated from the autoclaved litter kept at -12 C up to 60 days. From the nonautoclaved litter, viral RNA was detected up to 60 days and virus was isolated up to 14days. The present study indicated that APV could survive in built-up turkey litter up to 60 days postinoculation at a temperature of-12 C. PMID- 14562911 TI - Blindness due to retinal dysplasia in broiler chicks. AB - Thirty-three live 9-day-old broiler chicks were submitted for laboratory evaluation because of blindness. Blindness was observed in up to 1% of 25,000 birds from four different houses. All the chicks were from the same breeder source. Blindness was apparent when the chicks were 2-3 days old. Clinically, the blind chicks were smaller than their counterparts, were unable to find feed and water, and wandered aimlessly. Necropsy did not reveal any lesions. Two chicks examined clinically at 14 days of age lacked pupillary reflexes after light stimulation, and the anterior and posterior segments of the eye were normal. Microscopically, degeneration of photoreceptor cells characterized by vacuoles was evident at 9 days of age, and rosette formation of the retina, disorganization of retinal layers, synechia of the retina with mild inflammation in the choroid, and proliferation of the retinal pigment epithelial cells were evident by 15 days of age. Because the chicks came from the same breeder source and the incidence of blindness was low, a probable genetic etiology is suspected as the cause of blindness in this flock. PMID- 14562912 TI - Congenital skeletal abnormalities in a tawny owl chick (Strix aluco). AB - In this case report, we describe a tawny owl chick (Strix aluco) coming from a Wild Fauna Recovery Center with multiple congenital malformations in the limbs. The animal was unable to fly and showed marked malnutrition and poor general appearance. Physical, radiologic, and anatomic examinations showed osseous malformations including dislocation of radius and carpometacarpus with abnormal nonfunctional fixation of ligamentum propatagialis, absence of most parts of the bones of the manus in both wings, and twisted nonfused left tarsometatarsus with marked griphosis of digits. Routine toxicologic and pathologic examinations did not reveal a specific etiology. PMID- 14562913 TI - An outbreak of fowl cholera in ring-necked pheasants (Phasianus colchicus). AB - A total of 120 ring-necked pheasants from a 3000-bird flock in Zeeland, MI, died over a 3-day period. Clinical signs included sudden death, diarrhea, and limping. At necropsy, hepatomegaly with multifocal cream-colored foci randomly distributed throughout the parenchyma was observed in diseased birds. Additionally, the spleen was enlarged up to three times its normal size and had a marbled appearance. Microscopically, there was multifocal splenic and hepatic necrosis with intralesional rod-shaped bacteria. Pasteurlla multocida serotype 3/4 was isolated from liver and spleen. In this paper, we report an outbreak of acute fowl cholera in ring-necked pheasants. PMID- 14562914 TI - Releasing nitrogen from ammoniated lignin by white rot fungus cometabolizes environmental pollutants. AB - The nitrogen-modified lignocelluloses(NML) produced under oxic ammoniation was metabolized by white rot fungus, NH4(+) -N was released, NO3(-) -N concentration was decreased and total nitrogen loss was blocked within incubation period. During releasing nitrogen from the metabolism of NML, white rot fungus cometabolized recalcitrant environmental pollutants and showed higher degradation capability. Results indicated that this NML complex colonized by white rot fungus might be effective with economic feasibility when they are applied into the vast field ecosystem, it might stabilize NH4+ nitrogen flux and bioremediate the polluted environmental sites. PMID- 14562915 TI - Water solubility enhancements of PAHs by sodium castor oil sulfonate microemulsions. AB - Water solubility enhancements of naphthalene(Naph), phenantherene(Phen) and pyrene(Py) in sodium castor oil sulfonate(SCOS) microemulsions were evaluated. The apparent solubilities of PAHs are linearly proportional to the concentrations of SCOS microemulsion, and the enhancement extent by SCOS solutions is greater than that by ordinary surfactants on the basis of weight solubilization ratio(WSR). The logK(em) values of Naph, Phen, and Py are 3.13, 4.44 and 5.01 respectively, which are about the same as the logK(ow) values. At 5000 mg/L of SCOS concentration, the apparent solubilities are 8.80, 121, and 674 times as the intrinsic solubilities for Naph, Phen, and Py. The effects of inorganic ions and temperature on the solubilization of solutes are also investigated. The solubilization is improved with a moderate addition of Ca2+, Na+, NH4+ and the mixture of Na+, K+, Ca2+, Mg2+ and NH4+. WSR values are enhanced by 22.0% for Naph, 23.4% for Phen, and 24.6% for Py with temperature increasing by 5 degrees C. The results indicated that SCOS microemulsions improve the performance of the surfactant-enhanced remediation (SER) of soil, by increasing solubilities of organic pollutants and reducing the level of surfactant pollution and remediation expenses. PMID- 14562916 TI - Analysis of parental strain DNA fragments existing in GEMs-Fhhh. AB - There were 6 target DNA fragments of the three parental strains existing in the cell of GEMs (genetically engineered microorganism strain) Fhhh measured in this research by PCR(polymerase chain reaction). The determination showed that GEMs Fhhh contained all the 6 target DNA fragments, mnp1, mnp2, lip1, lip2, FLO1 and 16S rDNA, and had the molecular genetic stability. Meanwhile the PCR production of each parental strain could only had its target DNA fragments and was different from each other. It may illustrate that the technique of the inter-kingdom protoplast fusion for the construction of GEMs Fhhh through the process of intercellular gene recombination could be used as a reliable bioengineering technique to create the specific functional stain for the pollution control. PMID- 14562917 TI - Photoelectrocatalytic degradation of Rose Bengal. AB - An innovative photoelectrode, TiO2/Ti mesh electrode, was prepared by galvanostaticanodisation. The morphology and the crystalline texture of the TiO2 film on mesh electrode were examined by scanning electronic microscopy and Raman spectroscopy respectively. The examination results indicated that the structure and properties of the film depended on anodisation rate, and the anatase was the dominant component under the controlled experimental conditions. Degradation of Rose Bengal (RB) in photocatalytic (PC) and photoelectrocatalytic (PEC) reaction was investigated, the results demonstrated that electric biasing could improve the efficiency of photocatalytic reaction. The measurement results of TOC in photoelectrocatalytic degradation showed that the mineralisation of RB was complete relatively. The comparison between the degradation efficiency of RB in PEC process and that in aqueous TiO2 dispersion was conducted. The results showed that the apparent first-order rate constant of RB degradation in PEC process was larger than that in aqueous dispersion with 0.1%-0.3% TiO2 powder, but was smaller than that in aqueous dispersion with 1.0% TiO2. PMID- 14562918 TI - Reduction of nitrate from groundwater: powder catalysts and catalytic membrane. AB - The reduction of nitrate contaminant in groundwater has gained renewed and intensive attention due to the environmental problems and health risks. Catalytic denetrification presents one of the most promising approaches for the removal of nitrate from water. Catalytic nitrate reduction from water by powder catalysts and catalytic membrane in a batch reactor was studied. And the effects of the initial concentration, the amounts of catalyst, and the flux H2 on the nitrate reduction were also discussed. The results demonstrated that nitrate reduction activity and the selectivity to nitrogen gas were mainly controlled by diffusion limitations and the mass transfer of the reactants. The selectivity can improved while retaining a high catalytic activity under controlled diffusion condition or the intensification of the mass transfer, and a good reaction condition. The total nitrogen removal efficiency reached above 80%. Moreover, catalytic membrane can create a high effective gas/liquid/solid interface, and show a good selectivity to nitrogen in comparative with the powder catalyst, the selectivity to nitrogen was improved from 73.4% to 89.4%. PMID- 14562919 TI - Inhibition of nitrification in soil by metal diethyldithiocarbamates. AB - Nitrification acts as a key process in determining fertilizer use efficiency by crops as well as nitrogen losses from soils. Metal dithiocarbamates in addition to their pesticidal properties can also inhibit biological oxidation of ammonium(nitrification) in soil. Metal [M = V(III), Cr(III), Mn(II), Fe(III), Ni(II), Cu(II), Zn(II) and Co(II)] diethyldithiocarbamates (DEDTC) were synthesized by the reaction of sodium diethyldithiocarbamate with metal chloride in dichloromethane/water mixture. These metal diethyldithiocarbamates were screened for their ability to inhibit nitrification at different concentrations( 10 microg/g soil, 50 microg/g soil and 100 microg/g soil). With increasing concentration of the complex, capacity to retard nitrification increased but the extent of increase varied for different metals. At 100 microg/g soil, different complexes showed nitrification inhibition from 22.36% to 46.45% . Among the diethyldithiocarbamates tested, Zn(DEDTC)2 proved to be the most effective nitrification inhibitor at 100 microg/g soil. Manganese, iron and chromium diethyldithiocarbamates also proved to be effective nitrification inhibitors than the others at 100 microg/g soil. The order of percent nitrification inhibition in soil by metal diethyldithiocarbamates was: Zn(II) > Mn(II) > Fe(III) > Cr(III) > V(III) > Co(II) > Ni(II) > Cu(II). PMID- 14562920 TI - Estimating health damage cost from secondary sulfate particles--a case study of Hunan Province, China. AB - China's coal-dominated energy pattern has resulted in large amount of SO2 emissions. Estimate of the sulfur-related health damage cost is necessary to help perform systematic cost-benefit analysis and set national energy and emissions control priorities. Current researches were confined to gaseous SO2 in urban areas; however, secondary sulfate (SO4(2-)) particles can exert serious impact in a wider region. Based on the concept of "intake fraction", CALPUFF long-range dispersion model and 180 sample emission sources, multiple regression equation was obtained with good correlation( r = 0.85), which illustrates that populations were key parameters to determine intake fraction but source characteristics were insignificant. Based on the formula and the population distribution data, county level intake fractions were mapped for Hunan Province(range: 1.1 x 10(-6) - 3.2 x 10(-6)) of China. A combination of county-level SO2 emissions with the intake fractions yields a total 1.98 tons of sulfate(SO4(2-)) inhalation, and resulting total health damage cost to be 0.76(willingness to pay approach, WTP) or 0.16 (human capital approach, HC) billion USD in 1997, about 2.1% or 0.45% of GDP in Hunan in 1997. Average health damage cost per ton of SO2 emission is 930(WTP) or 200 USD(HC). The results demonstrated that more stringent regulation should be forced. PMID- 14562921 TI - Effects of nature organic matters and hydrated metal oxides on the anaerobic degradation of lindane, p, p '-DDT and HCB in sediments. AB - Effects of natural organic matters( NOM) and hydrated metal oxides( HMO) in sediments on the anaerobic degradation of gamma-666, p,p'-DDT and HCB were investigated by means of removing NOM and HMO in Liaohe River sediments sequentially. The results showed that the anaerobic degradation of gamma-666, p,p'-DDT and HCB followed pseudo-first-order kinetics in different sediments. But, the extents and rates of degradation were different, even the other conditions remained the same. Anaerobic degradation rates of gamma-666, p,p'-DDT and HCB were 0.020 d(-1), 0.009 d(-1) and 0.035 month(-1) respectively for the sediments without additional carbon resources. However, with addition of carbon resources, the anaerobic degradation rates of gamma-666, p,p'-DDT and HCB were 0.071 d(-1), 0.054 d(-1) and 0.088 month(-1) in the original sediments respectively. After removing NOM, the rates were decreased to 0.047 d(-1), 0.037 d(-1) and 0.066 month(-1); in the sediments removed NOM and HMO, the rates were increased to 0.067 d(-1) , 0.059 d(-1) and 0.086 month(-1). These results indicated that NOM in the sediments accelerated the anaerobic degradation of gamma-666, p,p'-DDT and HCB; the HMO inhibited the anaerobic degradation of gamma 666, p,p'-DDT and HCB. PMID- 14562923 TI - Comparison of decolorization of reactive microorganisms isolated from various sources. AB - Azo dyes are among the oldest man-made chemicals and they are still widely used in the textile, printing and the food industries. About 10% - 15% of the total dyes used in the industry is released into the environment during the manufacturing and usage. Some dyes and some of their N-substituted aromatic bio transformation products are toxic and/or carcinogenic and therefore these dyes are considered to be envionmental pollutants and health hazards. These azo dyes are degraded by physico-chemical and biological methods. Of these, biological methods are considered to be the most economical and efficient. In this work, attempts were made to degrade these dyes aerobically. The organisms which were efficient in degrading the following azo dyes-Red RB, Remazol Red, Remazol Blue, Remazol Violet, Remazol Yellow, Golden Yellow, Remazol Orange, Remazol Black- were isolated from three different sources viz., wastewater treatment plant, paper mill effluent treatment plant and tannery wastewater treatment plant. The efficiency of azo dye degradation by mixed cultures from each source was analyzed. It was found that mixed cultures from tannery treatment plant worked efficiently in decolorizing Remazol Red, Remazol Orange, Remazol Blue and Remazol Violet, while mixed cultures from the paper mill effluent worked efficiently in decolorizing Red RB, Golden Yellow and Remazol Yellow. The mixed cultures from wastewater treatment plant efficiently decolorized Remazol Black. PMID- 14562922 TI - Plants grown on sewage sludge in South China and its relevance to sludge stabilization and metal removal. AB - The production of sewage sludge in China has been increasing sharply in order to treat 40% of the municipal sewage in 2005 as planned by central government. The main sludge disposal method is landfill owing to heavy metal contamination, but it presents an attractive potential for agricultural land application. Experiments were carried out to study the simultaneous metal removal and sludge stabilization by plants. The sludge samples were collected from Datansha Wastewater Treatment Plant of Guangzhou, it contained excessive Cu and Zn compared with the Chinese National Standard for Agricultural Use of Sewage Sludge. Plants growing on sludge beds were investigated to follow their growth and metal uptake. 30 sludge plants were identified during 1 year's observation. A Zn high-accumulating and high growth rate plant(Alocasia macrorrhiza) was selected and grown on sludge beds in plots. The water, organic matter, heavy metals and nutrients contents, the E. coli number and the cress seed germination index were monitored for the sludge samples collected monthly. The plant growth parameters and its heavy metals contents were also determined. The sewage sludge treated by plants could be stabilized at about 5 months, the E. coli number was significantly decreased and the cress seed germination index attained 100%. Crop on sludge could ameliorate the sludge drying. The experiments are continuing to find out the appropriate plant combination for simultaneous sludge stabilization and metal removal for an acceptable period. Comparisons between the proposed processes and other methods for treating produced sludge such as composting, chemical andbacterial leaching were discussed. PMID- 14562924 TI - Immobilized chitosan as a selective absorbent for the nickel removal in water sample. AB - Method for preparation of chitosan immobilized on silica gel (CTS-silica) was described. The CTS-silica was used as absorbent for the absorption of nickel in water. The results showed that this absorbent had relatively high selectivity and strong affinity to nickel. The maximum absorption capacity for nickel can reach 667 mg/g of chitosan. Factors that affect the absorption capacity, such as pH, ion strength and the presence of calcium, EDTA and the mechanism of absorption were discussed in detail. The absorbent can be regenerated with acid and reused for several times. The recovery rate for nickel can reach 99.99%. This absorbent filled in a column can be used in nickel removal from wastewater and drinking water. PMID- 14562925 TI - Fluoride distribution and the effect of some ions along Alexandria coastal Mediterranean seawater of Egypt. AB - The coastal seawater of Mediterranean of Alexandria receives large amount of discharged waters containing industrial wastes, sewage, and agricultural and domestic drainage. Fluoride and some parameters were (chemical and physical) determined. The data gave indication that the content and the amount of the discharged water largely affect the chemical composition of the coastal water. Stepwise regression analysis was highly significant and the model was very fruitful, where the observed and calculated values were mostly concordant. This may indicated that there was a relation between fluoride content in coastal seawater and its content in the discharged water. PMID- 14562926 TI - Catalytic dechlorination of chlorobenzene in water by Pd/Fe bimetallic system. AB - Catalytic reductive dechlorination of monochlorobenzene(MCB) was carried out in the palladium/iron system. With low Pd loading (0.005%), 45% dechlorination efficiency was achieved within 5 h. Pd as catalyst accelerated the reductive dechlorination reaction. Dechlorination mechanism and kinetics were discussed. The reaction took place on the bimetal surface in a pseudo-first-order reaction, with the rate constant being 0.0071 min(-1) (K(SA) = 8.0 x 10(-3) L/(m2 x h)). The reduction product for MCB was benzene. PMID- 14562927 TI - United membrane biological reactor in the treatment of wastewater. AB - The united membrane biological reactor( UMBR) was studied for the treatment of some simulate and municipal wastewater. The removal efficiency for COD and turbidity are greater than 80% and 99% respectively. Effluent COD is less than 100 mg/L while turbidity less than 5. The removal of LAS in bath wastewater is greater than 70%. In treatment of dinning-hall wastewater, removal of fatty oil is greater than 90%, and its concentration in effluent is less than 5 mg/L. The match of biological reactor and the membrane separation component were calculated. The stable performance of wastewater treatment can be maintained by the optimization of operation conditions and the cleanout of membranes. PMID- 14562928 TI - Sedimentation retention basin utilization for best management practice. AB - Approaches to the artificial impoundment and theoretical design of sedimentation retention basin are reviewed with particular attention to best management practice (BMP) to control agriculture and surface runoff. Sediments retention basins are the small version of farm pond used where a criteria of farm pond is not met. Such basin traps the pollutants and suspended solids prior to entry into streams and lakes. The study is focused with special reference to the assessment and control of non-point source pollution (NPSP) from the sub-basin area of Tai Lake in the Xishan County of Wuxi City of China. The author suggested two different approaches to conduct this study including theoretical design for sedimentation retention basin and computation of flow, sediment transport and deposition during the artificial impoundment of retention basin for BMP's utilization. Theoretical design will provide a useful function as a first line defense against the movement of sediments and transport of pollutants into the Tai Lake while the assessment of sediments deposition will help to make its proper use and periodic cleanup. PMID- 14562929 TI - Temperature effect on aerobic denitrification and nitrification. AB - Nitrogen loss without organic removal in biofilter was observed and its possible reason was explained. A lower hydraulic loading could improve aerobic denitrification rate. Aerobic denitrification was seriously affected by low temperature(below 10 degrees C). However, nitrification rate remained high when the temperature dropped from 15 degrees C to 5 degrees C . It seemed the autotrophic biofilm in BAF could alleviate the adverse effect of low temperature. PMID- 14562930 TI - A novel approach to treat combined domestic wastewater and excess sludge in MBR. AB - Domestic wastewater was treated by combined anaerobic biofilm-aerobic membrane bioreactor(MBR) process, and part biomass MBR was withdrawn to treat with ozone, then the ozonated sludge was returned to anaerobic inlet. In aerobic MBR, MISS and DO were controlled at 3000-3500 mg/L and 0.8 mg/L respectively. Comparing the experimental results of two stages, it was noticed that ozonation did not affect the removal efficiency for organics but had a significant influence on the removals of NH3-N and TN. During the ozonation period of two months, no excess sludge was wasted, and a zero sludge yield was obtained. PMID- 14562931 TI - Control effects of p(epsilon) and pH on the generation and stability of chlorine dioxide. AB - A new method, without assistance of activity ratio diagram, was applied to construct the p(epsilon)-pH diagrams for chlorine system. The optimal pH range for generation of C1O2 by contacting Cl2 (g) directly with ClO2- solution is within pH 1.35-1.94, particularly within pH 1.35-4.00 only if minimizing the formation of Cl2. It is unachievable to synthesize pure ClO2 from the reaction of Cl2 and ClO2- . Conversely, ClO2 may be present a variation of stability in different waters owing to the changed p(epsilon) and pH. ClO2 could be relatively stable if not disproportionate into ClO3-, coexisting with ClO2- (p(epsilon) 17.63 and pH > 9.68), Cl2(pH < or = 0.92) or Cl- (pH 0.92-9.68). When chlorine system has already reached the ultimate equilibria, ClO2 is a stable species in strongly acid media. As the acidity decreases, ClO2 disproportionates into ClO3- and Cl2. Aqueous ClO2 is unstable within the normal pH range. This work initially, theoretically elucidates the generation and stability of ClO2 by way of the p(epsilon)-pH diagrams. PMID- 14562932 TI - Direct numerical simulation of the motion of circular pollutant particles in Newtonian fluid. AB - An improved implementation of distributed multiplier/fictitious domain method is presented for the direct numerical simulation of particulate flow. The key improvement is to replace a finite-element triangulation for the velocity and a "twice-coarser" triangulation for the pressure with a rectangular discretization for the velocity and pressure. For code validation, the sedimentation of a single particle in a two-dimensional channel was simulated. The results showed that the simulation is independent of the mesh size as well as the time step. The comparison between experimental data and this simulation showed that our code can give a more accurate simulation on the motion of particles than previous DLM code. The code was then applied to simulate the sedimentation of 600 particles in a rectangular box. The falling course is presented and discussed. At the same time, this simulation also demonstrates that the method presented in this paper can be used for solving the initial problems involving a lager number of particles exactly with computing durations kept at acceptable levels. PMID- 14562933 TI - Biodegradation of methanol vapor in a biofilter. AB - Volatile organic compounds (VOCs) are a new class of air pollutants posing threat to the environment. Newer technologies are being developed for their control among which biofiltration seem to be most attractive. Biofiltration of methanol vapor from air stream was evaluated in this study. Experimental investigations were conducted on a laboratory scale biofilter, containing mixture of compost and polystyrene inert particles as the filter materials. Mixed consortium of activated sludge was used as an inoculum. The continuous performance of biofilter for methanol removal was monitored for different concentrations and flow rates. The removal efficiencies decreased at higher concentrations and higher gas flow rates. A maximum elimination capacity of 85 g/(m3 x h) was achieved. The response of biofilter to upset loading operation showed that the biofilm in the biofilters was quite stable and quickly adapted to adverse operational conditions. PMID- 14562934 TI - Treatment of black liquor from the papermaking industry by acidification and reuse. AB - Two different kinds of black liquor from the papermaking industry were treated by acidification and reuse. The experimental parameters and conditions were discussed in detail. The experimental results indicated that the treatment process mentioned in this article is an effective process for the treatment of black liquor from the papermaking industry. By the treatment, the solid materials in black liquor are transferred into two by-products and the other components are reused or evaporated. Thus, no wastewater except some condensation water would be discharged in pulping process and the problem of pollution of black liquor would be effectively solved. PMID- 14562935 TI - Hydrological eco-service of rubber plantations in Hainan Island and its effect on local economic development. AB - The impacts of economic forest on global environmental change (GEC) are one of the hot issues in environmental study. Based on the 3-year observation data and 40-year climate data, GEC and analysis of the hydrological dynamic characteristics of rubber plantations and estimate of the water balance in the rubber plantations in Hainan Island were made. The results showed that the rainfall intercepted by the canopy of the plantations accounted for 11.45% of the annual rainfall, the total runoff for 23.71%, the total evaporation and transpiration for 63.24%, the soil moisture storage for 1.6%. Analysis of the 40 year rainfall data in the 19 counties of Hainan Island during 1951-1990 showed that the large-scale substitution of the natural vegetation with the rubber plantations had no significant effect on the local rainfall in Hainan Island. The main reasons are (1) 80% of the rainfall in Hainan is brought by typhoons; (2) the proportion of 11.6% rubber plantations in total forest coverage in Hainan is not enough to influence the local rainfall in Hainan Island; and (3) although the rubber plantation is artificial vegetation, it has the similar function to the tropical rain forest. Analysis of the total water resource and total GDP of Hainan in 1997 showed that the economic benefit resulted from the water resource was 1.0 RMB Yuan/m3. The value of hydrological of the rubber plantation in Hainan was 113.9 million RMB Yuan/3 when compared with the tropical rain forest. The paper reaches conclusion that the hydrological eco-service function of rubber plantation has been enhanced after transformed from natural vegetation, which includes the natural service and powerful social service. PMID- 14562936 TI - Sonochemical degradation of organophosphorus pesticide in dilute aqueous solutions. AB - Ultrasonic irradiation was found to accelerate the rate of hydrolysis of omethoate in aqueous solution over the pH range of 2-12. Process parameters studied include pH, steady-state temperature, concentration, and the type of gases. Greater than 96% hydrolysis was observed in 30 minutes through this process and the rate of destruction increased with the help of more soluble and low thermal inert gas. So with Krypton, omethoate was found to undergo rapid destruction as compared with Argon. In the presence of ultrasound, the observed first-order rate of hydrolysis of omethoate is found to be independent of pH. The formation of transient supercritical water (SCW) appears to be an important factor in the acceleration of chemical reactions in the presence of ultrasound. A detailed chemical reaction mechanism for omethoate destruction in water was formulated. Experimental results and theoretical kinetic mechanism demonstrated that the most of the omethoate undergo destruction inside the cavitating holes. A very less effect of temperature on the degradation of omethoate within a temperature range of 20-70 degrees C proves that a small quantity of omethoate undergoes secondary destruction in the bulk liquid. PMID- 14562937 TI - Soil CO2 flux in relation to dissolved organic carbon, soil temperature and moisture in a subtropical arable soil of China. AB - Soil CO2 emission from an arable soil was measured by closed chamber method to quantify year-round soil flux and to develop an equation to predict flux using soil temperature, dissolved organic carbon(DOC) and soil moisture content. Soil CO2 flux, soil temperature, DOC and soil moisture content were determined on selected days during the experiment from August 1999 to July 2000, at the Ecological Station of Red Soil, the Chinese Academy of Sciences, in a subtropical region of China. Soil CO2 fluxes were generally higher in summer and autumn than in winter and spring, and had a seasonal pattern more similar to soil temperature and DOC than soil moisture. The estimation was 2.23 kgCO2/(m2 x a) for average annual soil CO2 flux. Regressed separately, the reasons for soil flux variability were 86.6% from soil temperature, 58.8% from DOC, and 26.3% from soil moisture, respectively. Regressed jointly, a multiple equation was developed by the above three variables that explained approximately 85.2% of the flux variance, however by stepwise regression, soil temperature was the dominant affecting soil flux. Based on the exponential equation developed from soil temperature, the predicted annual flux was 2.49 kgCO2/(m2 x a), and essentially equal to the measured one. It is suggested the exponential relationship between soil flux and soil temperature could be used for accurately predicting soil CO2 flux from arable soil in subtropical regions of China. PMID- 14562939 TI - Understanding prosthodontics--where did it all go wrong? AB - This paper addresses the anatomical and physiological effects of tooth loss on the patient and, further, audits the teaching of prosthodontics over the last 30 years. There is an abundance of evidence to outline changes that occur post extraction related to morphological changes in the maxilla, the mandible and combined changes in inter-maxillary relations. In addition to the knowledge of patterns of bone loss, we are also now well aware that there is evidence to suggest that planned retention of roots and/or the placement of implants can help reduce post-extraction bone loss. Awareness by clinicians of the anatomical, physiological and psychological effects of edentulousness, is essential if empathy is to be struck with the patient. Equally, patients being rendered edentulous must be made aware of these factors if true informed consent is to be gained. New graduates are now less well equipped to practice complete denture prosthodontics than, say, 10 years ago and this is a consequence of three factors: crowded curricula, diminution in laboratory hours and the time a dental student spends in clinical prosthodontics. PMID- 14562938 TI - Tooth loss and dentures: patients' perspectives. AB - OBJECTIVE: Despite dramatic improvements in tooth retention around the world, a substantial proportion of older adults have lost natural teeth and many wear removable partial and complete dentures. Problems associated with tooth loss and denture wearing remain important in the context of global ageing. The purpose of this paper is to examine the effects of tooth loss and denture wearing on their day-to-day lives from the patient perspective. DESIGN: Cross-sectional study. SETTING: Greater Boston area, USA. PARTICIPANTS: Community-dwelling older men. METHODS: Brief examination and survey. MAIN OUTCOME MEASURES: Self-reported oral health measures including the single-item self-rating of oral health, the Oral Health Impact Profile (OHIP), the Geriatric Oral Health Assessment Index (GOHAI), Oral-Health-related Quality of Life (OHQOL) and a newly-developed short-form instrument (the DELTA). RESULTS: Men with > or = 25 teeth had better self-rated oral health by all measures. The new, brief DELTA differentiates between dentition/denture groupings as well as or better than existing instruments. Over 80% of men with > or = 25 teeth rated their oral health as excellent, very good or good, compared with 70% of men with no teeth (and dentures) and 54% of men with 1-24 teeth. Avoidance of certain foods discriminates well between dentition groups. To a lesser extent, difficulty with relaxation, pain and distress, and avoidance of going out are associated with tooth loss and/or denture wearing. PMID- 14562940 TI - Complete denture success for patients and dentists. AB - The huge prevalence of tooth decay and the gradual ageing of populations in the developed world have combined to produce a steady increase in the need for complete denture treatment. To achieve successful treatment, it is vital to consider a number of issues throughout the denture process. Before treatment even begins, the patient's motivation for denture treatment and emotional attitude towards dentures must be evaluated. Patients will thereby gain realistic expectations of what can and cannot be achieved, and dentists will understand what the patient really wants. Afterwards, the dentist must work with the patient to help achieve success in difficult areas such as neuromuscular control, mastication and speech. By learning to handle patient complaints positively, dentists can educate patients to solve their own problems in future. Finally, patients must be informed that continued success depends on regular denture maintenance at home combined with periodic consultation with the dentist. PMID- 14562941 TI - Targeted enlargement of surveillance activities. PMID- 14562942 TI - Resistance to extended-spectrum cephalosporins and mortality in patients with Citrobacter freundii bacteremia. AB - BACKGROUND: This study was performed to characterize the clinical features and to identify the risk factors for resistance to extended-spectrum cephalosporins (ESCs) and for mortality in patients with Citrobacter freundii bacteremia. PATIENTS AND METHODS: 105 patients (aged > or = 15 years) with C. freundii bacteremia in 1991-2000 were retrospectively analyzed. RESULTS: Nosocomial acquisition was identified in 78.1% of the patients. Hepatic, biliary and pancreatic disease was the most common underlying disease (65.7%) and the biliary tract was the most common site of infection (50.5%). The overall resistance rate to ESCs was 59.0% and was significantly associated with hepatic, biliary and pancreatic disease, recent surgery and procedure, biliary drainage catheter and previous antibiotic therapy in univariate analysis. However, only previous antibiotic therapy with ESCs (OR = 5.0, 95% CI 1.6-15.7, p = 0.006) and recent surgery or procedure (OR = 3.1, 95% CI 1.1-8.4, p = 0.03) were strong, independent risk factors in multivariate analysis. Mortality directly related to C. freundii bacteremia was 21.9% and there was no difference between cases with resistance and susceptibility to ESCs (19.4% vs 25.6%; p = 0.45). Mortality was significantly associated with rapidly fatal or ultimately fatal underlying disease, a solid tumor, septic shock and polymicrobial bacteremia in univariate analysis. Among patients who had therapeutic surgical procedures, mortality was lower (4.5%, p = 0.04). Multivariate analysis revealed rapidly or ultimately fatal disease, septic shock and polymicrobial bacteremia as independent prognostic factors. CONCLUSION: Biliary infection was the leading cause of C. freundii bacteremia. Previous antibiotic therapy, especially with ESCs, frequently predisposed for resistance to these antibiotics. However, resistance to ESCs was not associated with increased mortality. PMID- 14562943 TI - Design of a surveillance system of antibiotic use and bacterial resistance in German intensive care units (SARI). AB - BACKGROUND: Data on antibiotic consumption and bacterial resistance are important for benchmarking, ensuring quality of antibiotic treatment and helping to understand the relationship between the use of antibiotics and the emergence of resistance. METHODS: The SARI project is an ecological study that has established laboratory-based surveillance in German intensive care units (ICU). Resistance rates of 13 sentinel pathogens are reported and certain alert organisms are sent for genotyping and retesting of antimicrobial resistance. RESULTS: The project, initiated in February 2000, now includes 35 ICUs generating a total of 266,013 patient days, 354,356 defined daily doses (DDD) and providing susceptibility data on 21,354 isolates. Pooled antibiotic usage density (AD = DDD/1,000 patient days) was highest for penicillins with lactamase inhibitor (AD 338.3) followed by quinolones (AD 155.5) and second-generation cephalosporins (AD 124.6). Total AD was calculated as 1,337 DDD/1,000 patient days. Resistance rates (RR) for laboratories testing according to the German Industrial Standard (DIN) were 19.3% for methicillin-resistant Staphylococcus aureus (MRSA), 9.5% for ciprofloxacin resistant Escherichia coli and 25.4% for imipenem-resistant Pseudomonas aeruginosa. 40% of the laboratories did not identify the extended spectrum beta lactamase production of a Klebsiella pneumoniae strain. CONCLUSION: Focusing on German ICUs, the SARI surveillance system provides a concept that produces a benchmark for the link between antibiotic resistance and consumption. PMID- 14562944 TI - Virulence characteristics of Escherichia coli strains causing asymptomatic bacteriuria. AB - BACKGROUND: The objective of this study was to examine the expression of Escherichia coli virulence-associated factors among the strains isolated from a group of women with a history of recurrent urinary tract infections (UTIs), in whom asymptomatic bacteriuria (ABU) was detected at follow-up, and from a group of children without a history of previous UTI, in whom ABU was detected during the screening. Possible differences between the virulence potential of these strains were investigated. MATERIALS AND METHODS: Hemolysin production, the ability to adhere to Buffalo green monkey cell line and hemagglutination (HA) ability of the ABU-associated E. coli strains were tested. E. coli strains isolated from patients with acute recurrent UTIs served as a comparison. RESULTS: The well-known low virulence of strains isolated from patients with ABU was demonstrated. In contrast to strains isolated from recurrent uncomplicated UTIs, the ABU-associated strains were mostly nonhemolytic (75%), nonadherent (70%) and lacked HA ability (61%). HA ability was significantly more common among the strains isolated from children without a history of UTI than among the strains isolated from women with recurrent UTIs (chi2 = 9.97, p < 0.01), whereas the adherence and hemolytic abilities did not differ between the two ABU groups. CONCLUSION: A further prospective study is needed to determine whether the HA ability is the predictor of subsequent symptomatic UTI. PMID- 14562945 TI - Specific hepatitis B vaccine therapy in inactive HBsAg carriers: a randomized controlled trial. AB - BACKGROUND: Hepatitis B virus (HBV) vaccine therapy has two major areas of application: for preventive purposes and for treating patients with chronic hepatitis B. This study aimed to investigate the effect of therapeutic vaccination of inactive hepatitis B surface antigen (HbsAg) carriers using a recombinant hepatitis B vaccine in a randomized-controlled study. PATIENTS AND METHODS: The 71 studied patients had never received prior antiviral therapies, were anti-HBe positive, had undetectable HBV-DNA and persistently normal alanine transaminase levels. 31 patients were given three 20 mg intramuscular injections of a preS2/S vaccine (GenHevac-B) on days 0, 30 and 60 and the remaining 40 patients were included in the control group. The efficacy of vaccination was evaluated by testing for HBsAg seroconversion to anti-HBs. Post-vaccination follow-up was for 12 months after the first dose. RESULTS: At the end of the follow-up, three out of 31 patients (10%) who received vaccine therapy were able to clear HBsAg from their sera and concomitantly develop anti-HBs antibodies. In contrast, none of the 40 control patients who did not received vaccine therapy had decreased their levels of HBsAg or elicited anti-HBs antibodies (p = 0.079). In three vaccinated patients serum HBsAg became undetectable approximately by the 3rd month of vaccine therapy and HBsAg seroconversion was seen to be durable in all patients in the follow-up period. CONCLUSION: This study offers the first direct evidence, based on a controlled study, that the recombinant HBV vaccine has no great effect in enhancing the rate of HBsAg seroconversion in inactive HBsAg carriers. More efficient strategies, such as an increase in the dose and number of immunizations, should be evaluated further in large controlled trials. PMID- 14562946 TI - Prevalence and persistence of TT virus DNA in HIV1-infected individuals. AB - BACKGROUND: TT virus (TTV) is a recently discovered virus with a high DNA prevalence in different populations. Its role in pathogenesis is uncertain, particularly in immunocompromised patients. PATIENTS AND METHODS: Prevalence of TTV-DNA was evaluated in a cohort of HIV-infected patients and in blood donors by nested PCR, using two different primer sets: T primers, derived from the open reading frame ORF1 region N22; B primers, derived from the untranslated region (UTR). Samples positive using T primers were also tested for TTV-DNA in peripheral blood mononuclear cells (PBMC) and followed up every 4 months. RESULTS: The overall prevalence of TTV-DNA in HIV-infected patients was 37/376 (9.8%) using T primers and 223/333 (67%) using B primers; prevalence was higher in males (167/237, 70.5% vs 56/96, 58.3%; p = 0.033) and sub-Saharan Africans (22/23, 95.6% vs 201/310, 64.8% in other areas). Discordance was also observed in blood donors: 3.8% prevalence using T primers and 51.4% using B primers (also higher in males: 57% vs 37%, p = 0.056). TTV-DNA was detected in PBMC in 20/23 (87%) TTV-positive sera. Two-thirds of the serum samples remained positive over a 2-year follow-up period. CONCLUSION: TTV-DNA prevalence is higher when detected with primers derived from the UTR region and was highest in male and HIV-infected sub-Saharan Africans. TTV-DNA is frequently isolated in PBMC and chronic infection is common. PMID- 14562947 TI - HIV and hepatitis C virus: facts and controversies. AB - Hepatitis C virus (HCV) infection occurs in about one-third of HIV-seropositive patients and in about 90% of HIV-positive drug abusers. After the introduction of highly active antiretroviral therapy (HAART) and the subsequent reduction in mortality from opportunistic infections, HCV-related liver failure has become a frequent cause of death in HIV-positive patients. In HIV-seropositive patients, the course of HCV infection is accelerated and there is evidence that HCV is an important factor for HIV progression. Consequently, it is important to establish the appropriate treatment for HCV infection in HIV-seropositive patients. This review examines the epidemiology, physiopathology, diagnostics and treatment of HIV/HCV coinfection with particular regard to the impact of HAART. PMID- 14562948 TI - Successful treatment of pulmonary invasive aspergillosis with voriconazole in patients who failed conventional therapy. AB - BACKGROUND: The incidence of fungal infections, including those due to Aspergillosis species has continued to increase in recent years. Invasive aspergillosis remains an important cause of morbidity and mortality, despite therapeutics interventions. PATIENTS AND METHODS: We reported five cases of invasive pulmonary aspergillosis treated with voriconazole failing to respond to conventional treatments. RESULTS: The clinical and radiological resolution of pulmonary aspergillosis reported in these cases following therapy with voriconazole is remarkable, considering the infections had proved refractory to standard antifungal therapies. Long-term therapy (in two cases > or = 1 year, in one case 6 months) was very well tolerated by patients who were unable to tolerate other antifungal agents. CONCLUSION: Therapy with voriconazole offers a new therapeutic option for otherwise difficult-to-treat infections and the potential to significantly improve the management of Aspergillosis infections. PMID- 14562949 TI - Maternal carriage and antimicrobial resistance profile of group B Streptococcus. AB - BACKGROUND: The aim of this study was to determine the prevalence of group B Streptococcus (GBS) colonization and to evaluate the antimicrobial resistance profile in women in the third trimester of pregnancy. MATERIALS AND METHODS: A total of 310 pregnant women, referred in weeks 35 to 37 of gestation, were screened for GBS colonization during a 10-month period. Samples were collected from the vagina and the rectum. RESULTS: The colonization rate was 10.6% and 22 women (66.7%) had both positive vaginal and rectal cultures. Rates of GBS colonization were significantly lower in patients aged 24 years or older and in those with a third or later pregnancy. None of the isolates were resistant to penicillin and ampicillin, whereas 21.2% and 9.1% showed resistance to erythromycin and clindamycin, respectively. CONCLUSION: Screening and antimicrobial susceptibility testing of GBS during pregnancy are important to guide appropriate therapy. PMID- 14562950 TI - Cerebrospinal fluid glucose levels and sensorineural hearing loss in bacterial meningitis. AB - BACKGROUND: Low cerebrospinal fluid (CSF) glucose levels have been associated with sensorineural hearing loss (SNHL) in bacterial meningitis but systematic investigations are lacking. PATIENTS AND METHODS: A retrospective case control study was conducted comparing CSF glucose levels in 47 cases with and 145 controls without SNHL following bacterial meningitis. RESULTS: The mean CSF glucose level (standard deviation [SD]) was 1.3 mmol/l (1.2) in cases and 2.5 mmol/l (1.6) in controls (p < 0.001). The difference between the groups was significant for Streptococcus pneumoniae and Haemophilus influenzae but not for Neisseria meningitidis infection. It was independent of the delay of treatment if this was greater than 12 h. This delay was significantly longer in cases compared to controls. CONCLUSION: In bacterial meningitis other than that caused by N. meningitidis and independent of the duration of symptoms prior to diagnosis, CSF glucose levels were significantly lower in patients developing a SNHL compared to controls. PMID- 14562951 TI - Streptococcus pneumoniae as an uncommon cause of superinfected pancreatic pseudocysts. AB - We report a patient with pancreatic pseudocysts that were superinfected with Streptococcus pneumoniae. The literature on the prevalence of superinfection of pancreatic tissue by S. pneumoniae, as well as on its prophylaxis and treatment, is reviewed. In addition, a possible pathophysiologic pathway is discussed. PMID- 14562953 TI - Monthly recurrent herpes simplex virus blepharitis in a boy for more than 10 years. AB - Ocular herpes simplex virus (HSV) infection is generally accepted to be a unilateral disease and simultaneous bilateral recurrent ocular HSV disease is uncommon. Recurrent ocular herpes was generally thought to be characterized by corneal involvement. We here report an 11-year-old boy with monthly bilateral recurrent HSV type 1 blepharitis for more than 10 years. He had a general normal immunological examination. Only supportive or topical acyclovir ointment treatment proved adequate for controlling the monthly recurrent disease without corneal involvement or other sequelae to date. The case highlights the unusual presentation, general normal immune function, clinical course and treatment opinion for recurrent HSV blepharitis. PMID- 14562952 TI - Drug hypersensitivity syndrome to carbamazepine and human herpes virus 6 infection: case report and literature review. AB - We describe a patient with a drug-induced hypersensitivity syndrome to carbamazepine and a concomitant active infection with human herpes virus 6 (HHV 6). The potential role of HHV-6 regarding the drug-induced hypersensitivity syndrome is discussed and the main clinical features of this potentially fatal adverse drug reaction are highlighted. PMID- 14562954 TI - Fatal malaria in a German couple returning from Burkina Faso. AB - We report on a German couple found dead at home 7 days after returning from Burkina Faso. Postmortem evaluation revealed Plasmodium falciparum infection with a parasitemia of approximately 80% in both cases. No pathological findings of the organs were present at autopsy except moderate splenomegaly in both cases and myocarditis in one case. Typical findings of fatal malaria like petechial hemorrhages of the brain or other organs, or sequestration of parasitized erythrocytes in venules and capillaries were absent. Lack of sequestration with excessive hyperparasitemia leading to systemic hypoxemia and host cytokine release may have contributed to the rapid fatal course in these nonimmune patients. PMID- 14562955 TI - European Commission approves Fuzeon following FDA approval--first HIV-fusion inhibitor in fight against HIV. PMID- 14562956 TI - Voriconazole--superior in antimycotic treatment. PMID- 14562958 TI - The evolutionary history of Shigella and enteroinvasive Escherichia coli revised. AB - In Shigella and enteroinvasive Escherichia coli (EIEC), the etiologic agents of shigellosis in humans, the determinants responsible for entry of bacteria into and dissemination within epithelial cells are encoded by a virulence plasmid. To understand the evolution of the association between the virulence plasmid and the chromosome, we performed a phylogenetic analysis using the sequences of four chromosomal genes (trpA, trpB, pabB, and putP) and three virulence plasmid genes (ipaB, ipaD, and icsA) of a collection of 51 Shigella and EIEC strains. The phylogenetic tree derived from chromosomal genes showed a typical "star" phylogeny, indicating a fast diversification of Shigella and EIEC groups. Phylogenetic groups obtained from the chromosomal and plasmidic genes were similar, suggesting that the virulence plasmid and the chromosome share similar evolutionary histories. The few incongruences between the trees could be attributed to exchanges of fragments of different plasmids and not to the transfer of an entire plasmid. This indicates that the virulence plasmid was not transferred between the different Shigella and EIEC groups. These data support a model of evolution in which the acquisition of the virulence plasmid in an ancestral E. coli strain preceded the diversification by radiation of all Shigella and EIEC groups, which led to highly diversified but highly specialized pathogenic groups. PMID- 14562957 TI - Adaptative value of a PKC-PKI55 feedback loop of inhibition that prevents the kinase's deregulation. AB - A 168-bp amplification product was obtained in RT-PCR experiments using a degenerate oligonucleotide designed on a five-amino acid sequence of IN, a 7-kDa protein, previously characterized as a PKC inhibitor. It was included in the coding ORF of the 1530-bp-long IMAGE clone ID 38900 (accession numbers R51337 and R51448) that produces a translation product of 6.5 kDa. The translation of the ORF conceptual reading frame allowed the preparation of the synthetic protein PKI55, which was found to inhibit and degrade both untreated nPKC d isozymes and activated cPKC isozymes. The PKI55 gene is localized in chromosome 2q35. The Repeat Maskers output showed a 533-bp-long LTR32/ERVL segment that included the PKI55 coding sequence and a complete regulatory region. The coding sequence and the structure of PKI55 were detected in a brain cDNA of Macaca fascicularis (diverged from human lineages about 25 Myr ago). Three other human genes with over 60% identities with PKI55 were identified in three different loci (i.e., chromosomes 10, 15, and 20.) Synthesis of PKI55 was stimulated by PKC activation. A feedback loop of inhibition is established. When the PKCs are overactivated, PKI55 induces degradation of the enzyme and prevents the isozyme overexpression implicated in a number of important diseases including cancer, diabetes, and disorders of the immune system. The presence of the PKI55 sequence in Macaca fascicularis as well as in human chromosomes 10, 15, and 20 indicates a selective advantage for the PKI55 sequence and the adaptive value of the feedback mechanism. PMID- 14562959 TI - Analysis of sequence periodicity in E. coli proteins: empirical investigation of the "duplication and divergence" theory of protein evolution. AB - Periodicity was quantified in 4289 Escherichia coli K12 confirmed and putative protein sequences, using a simple chi-square technique previously shown to reveal triplet period periodicity in coding DNA. Periodicities were calculated from period n = 2 to period n = 50 in nine different alphabetic representations of the proteins. By comparison with a randomly generated proteome of the same compositional content, the E. coli proteome does not contain a significant excess of periodic proteins. However, 60 proteins do appear to be significantly periodic in at least one alphabetic representation, after Bonferroni correction, at p < 0.01, and 30 at p < 0.001. These are compared with significantly periodic proteins of solved three-dimensional structure, detected by an identical analysis of the sequences from a protein structure database. It is concluded that there is no evidence for the presence of a proteome-wide quasi-periodicity as predicted by the "duplication and divergence" model of protein evolution and that the major periodicity detected is a consequence of the repetitive tendencies within alpha helices. However, it is not possible to explain all sequence periodicities in terms of observable secondary structure, as in cases where sequence periodicity can be compared to solved structure, there is often no structural regularity that would provide an obvious explanation in terms of natural selection on protein function. PMID- 14562960 TI - Reduced polymorphism in the chimpanzee semen coagulating protein, semenogelin I. AB - The semen of many primate species coagulates into a mating plug believed to prevent the sperm of subsequent mating events from accessing the ova. The texture of the coagulum varies among species: from a semisoft mass in humans to a firm plug in chimpanzees. In humans, a component of the coagulum, semenogelin I, also inhibits sperm motility. We tested the hypothesis that polymorphism and divergence at semenogelin I differ among hominoid species with different mating systems. Sequence data for the semenogelin I locus were obtained from 12 humans, 10 chimpanzees, 7 gorillas, and 1 bonobo. Mitochondrial D-loop data were collected from a subset of individuals to assess levels of variation at an unlinked locus. HKA tests using D-loop sequence data revealed a significant reduction of polymorphism at semenogelin I in chimpanzees, consistent with predictions of a selective sweep at this locus. This result was supported by independent HKA tests using polymorphism data from a putatively neutral locus from the literature. Humans show a similar trend toward reduced polymorphism, although HKA tests were only marginally significant. Gorilla sequence data show evidence of functional loss at the semenogelin I locus, indicated by stop codons within the putative open reading frame as well as high levels of polymorphism. Elevated Ka/Ks ratios within the Pan-Homo clade suggest a history of positive selection at semenogelin I. Our results suggest that there is a positive relationship between the intensity of sperm competition in a species and the strength of positive Darwinian selection on the seminal protein semenogelin I. PMID- 14562961 TI - Exploring the evolutionary history of the alcohol dehydrogenase gene (Adh) duplication in species of the family tephritidae. AB - In the olive fruit fly Bactrocera oleae and the med fly Ceratitis capitata previous studies have shown the existence of two Adh genes in each species. This observation, in combination with the former finding that various Drosophila species of virilis and repleta group encode two isozymes of ADH which are the result of a gene duplication, challenged us to address a scenario dealing with the evolutionary history of the Adh gene duplication in Tephritidae. In our lab we proceeded to the cloning and sequence analysis of Adh genes from more tephritid species, a prerequisite for further study of this issue. Here we show that phylogenetic trees produced from either the nucleotide or the amino acid sequences of 14 tephritid Adh genes consisted of two main clusters, with Adh sequences of the same "type" grouping together (i.e., Adh1 sequences form a cluster and Adh2 sequences form a second one), as expected if there was one duplication event before speciation within the family Tephritidae. We used the amount of divergence between the two isozymic forms of Adh of the species carrying both Adh1 and Adh2 genes to obtain an estimate of the age of the duplication event. Interestingly, our data again support the hypothesis that the duplication of an ancestral Adh single gene in the family Tephritidae occurred before the emergence of the genera Bactrocera and Ceratitis, thus suggesting that Adh duplication was based on a prespeciation rather than a postspeciation event that might have involved two independent duplication events, one in each of the two genera. PMID- 14562962 TI - Amino acid sequences and distribution of high-potential iron-sulfur proteins that donate electrons to the photosynthetic reaction center in phototropic proteobacteria. AB - High-potential iron-sulfur protein (HiPIP) has recently been shown to function as a soluble mediator in photosynthetic electron transfer between the cytochrome bc1 complex and the reaction-center bacteriochlorophyll in some species of phototrophic proteobacteria, a role traditionally assigned to cytochrome c2. For those species that produce more than one high-potential electron carrier, it is unclear which protein functions in cyclic electron transfer and what characteristics determine reactivity. To establish how widespread the phenomenon of multiple electron donors might be, we have studied the electron transfer protein composition of a number of phototrophic proteobacterial species. Based upon the distribution of electron transfer proteins alone, we found that HiPIP is likely to be the electron carrier of choice in the purple sulfur bacteria in the families Chromatiaceae and Ectothiorhodospiraceae, but the majority of purple nonsulfur bacteria are likely to utilize cytochrome c2. We have identified several new species of phototrophic proteobacteria that may use HiPIP as electron donor and a few that may use cytochromes c other than c2. We have determined the amino acid sequences of 14 new HiPIPs and have compared their structures. There is a minimum of three sequence categories of HiPIP based upon major insertions and deletions which approximate the three families of phototrophic proteobacteria and each of them can be further subdivided prior to construction of a phylogenetic tree. The comparison of relationships based upon HiPIP and RNA revealed several discrepancies. PMID- 14562963 TI - Phylogenetic analysis of the cytochrome P450 3 (CYP3) gene family. AB - Cytochrome P450 genes (CYP) constitute a superfamily with members known from the Bacteria, Archaea, and Eukarya. The CYP3 gene family includes the CYP3A and CYP3B subfamilies. Members of the CYP3A subfamily represent the dominant CYP forms expressed in the digestive and respiratory tracts of vertebrates. The CYP3A enzymes metabolize a wide variety of chemically diverse lipophilic organic compounds. To understand vertebrate CYP3 diversity better, we determined the killifish (Fundulus heteroclitus) CYP3A30 and CYP3A56 and the ball python (Python regius) CYP3A42 sequences. We performed phylogenetic analyses of 45 vertebrate CYP3 amino acid sequences using a Bayesian approach. Our analyses indicate that teleost, diapsid, and mammalian CYP3A genes have undergone independent diversification and that the ancestral vertebrate genome contained a single CYP3A gene. Most CYP3A diversity is the product of recent gene duplication events. There is strong support for placement of the guinea pig CYP3A genes within the rodent CYP3A diversification. The rat, mouse, and hamster CYP3A genes are mixed among several rodent CYP3A subclades, indicative of a complex history involving speciation and gene duplication. PMID- 14562964 TI - Widespread adaptive evolution in the human immunodeficiency virus type 1 genome. AB - We investigated variable selective pressures among amino acid sites in HIV-1 genes. Selective pressure at the amino acid level was measured by using the nonsynonymous/synonymous substitution rate ratio (omega = dN/dS). To identify amino acid sites under positive selection with omega > 1, we applied maximum likelihood models that allow variable omega ratios among sites to analyze genomic sequences of 26 HIV-1 lineages including subtypes A, B, and C. Likelihood ratio tests detected sites under positive selection in each of the major genes in the genome: env, gag, pol, vif, and vpr. Positive selection was also detected in nef, tat, and vpu, although those genes are very small. The majority of positive selection sites is located in gp160. Positive selection was not detected if omega was estimated as an average across all sites, indicating the lack of power of the averaging approach. Candidate positive selection sites were mapped onto the available protein tertiary structures and immunogenic epitopes. We measured the physiochemical properties of amino acids and found that those at positive selection sites were more diverse than those at variable sites. Furthermore, amino acid residues at exposed positive selection sites were more physiochemically diverse than at buried positive selection sites. Our results demonstrate genomewide diversifying selection acting on the HIV-1. PMID- 14562965 TI - Retention of a duplicate gene through changes in subcellular targeting: an electron transport protein homologue localizes to the golgi. AB - Cytochrome c oxidase (COX), the terminal enzyme complex of the electron transport chain, contains 13 subunits, 3 encoded by mitochondrial DNA and 10 by nuclear. Several of the nuclear subunits, including subunit VIIa, are known to have two tissue- and development-specific isoforms in mammals. A recently identified third member of the gene family, COX7AR, encodes a protein previously thought to function in mitochondria. However, observation of fluorescent pCOX7AR C-terminal fusion proteins in HeLa cells showed that pCOX7AR is localized to the Golgi apparatus. Sequence analyses indicate that the duplication of COX7AR occurred prior to the origin of the Euteleostomi (bony vertebrates) and that pCOX7AR is more highly conserved than the two other isoforms. These results indicate that, after gene duplication and modification of the mitochondrial targeting signal, pCOX7AR was evolutionarily altered to a new and apparently important function in the Golgi. These results also suggest that predictions of function from homology can be misleading and show that specialization and modification of subcellular localization are similar to cis-element subfunctionalization. In cis-element subfunctionalization, complementary null mutations occur to the cis-elements of the descendents of a gene duplication, causing both descendent genes to be obligate. In the process described in this paper, which could be termed subcellular subfunctionalization, complementary null mutations can occur to the subcellular localization signals of the descendants of a gene duplication, causing both descendent genes to be similarly obligate. Noncomplementary null mutations could also uncover an alternate localization, which is the more likely case for pCOX7AR. PMID- 14562966 TI - Phylogenetic analysis of new plant myosin sequences. AB - We have sampled a large number of plant taxa, ranging from brown algae to angiosperms, for the presence of myosin sequences. Using phylogenetic analysis, we show that all but two of the new plant myosin sequences fall into two of three preexisting myosin classes. We identified two outlying sequences, which do not fall into any preexisting myosin class. Additionally, all genomic sequences encoding class XI myosins contain an intron in the region studied, suggesting that this genomic region has been conserved over at least 1 billion years of plant evolution. With these data, we can rapidly and consistently classify partial myosin sequences from plants. Our data show that plant myosins do not have clear orthologues in other kingdoms, providing interesting insights into the diversification of myosins. PMID- 14562967 TI - Endovascular therapy of traumatic vascular lesions of the head and neck. AB - Pseudoaneurysm and fistula formation are well-documented complications of arterial vascular injury and may be associated with significant morbidity and mortality. The purpose of this manuscript is to review the presentation and therapy of patients with traumatic vascular injuries of the head and neck. Eight patients were admitted to a Level 1 Trauma Center and diagnostic angiography of the carotid artery and vertebral circulation was performed. The mechanisms of injury included motor vehicle accident, gunshot wound, stab wound and aggravated assault. Cause of trauma, vascular lesion, endovascular therapy and outcome were analyzed retrospectively. The angiographic findings, clinical presentation and hospital course were reviewed. There were eight patients, seven males and one female, aged 17-65. Four patients (50%) had multiple lesions; four had pseudoaneurysms, two with fistula formation and two with active arterial hemorrhage. A total of 17 lesions were embolized using coils. Polyvinyl Alcohol (PVA), Gelfoam or a combination. Two of the 17 lesions received stents. Six of the eight patients remained clinically improved or stable at varying follow-up intervals. One of the four patients who presented with penetrating trauma and neurological deficits had resolution of right hemiplegia at the 8th month follow up. One of the four patients who sustained blunt trauma and carotid-cavernous fistula presented with a new pseudoaneurysm at the 2-month post-embolization follow-up. The evolution of diagnostic neuroangiographic techniques provides opportunities for endovascular therapy of traumatic vascular lesions of the head and neck that are minimally invasive, attractive options in selected cases. PMID- 14562968 TI - Vena cava filter behavior and endovascular response: an experimental in vivo study. AB - PURPOSE: To evaluate the behavior and endovascular response of a new nitinol permanent vena cava filter, the TrapEase. METHODS: Percutaneous implantation of the filter was performed in six goats, with inferior vena cava (IVC) diameter close to that of man. Radiologic data concerning the IVC, filter diameter, patency and stability were collected. At 2, 4, 20 and 26 weeks post-implantation, histopathologic analysis of the IVC wall was performed at the site of filter distension, and distal and proximal to the filter. RESULTS: All filters remained patent. There was no migration and no signs of biological incompatibility. Signs of neointimalization were seen at 2 weeks, with well-developed neointima at 4 weeks. No acute vessel wall perforation was detected by cavography at implantation. During follow-up histologic analysis at 26 weeks, perforation of some of the small fixation barbs was seen, causing minimal damage to the vessel wall and adjacent organ tissue without impairing organ function. These events were well tolerated, probably due to the gradual nature of the penetration of fixation barbs allowing reactive fibrous tissue development. At 26 weeks the parallel filter struts were well covered with neointima and did not perforate the vessel wall. There were no complications associated with the filter implantation. CONCLUSIONS: The TrapEase vena cava filter was well tolerated and is suitable for incorporation into the IVC wall of healthy animals without any apparent deleterious reaction due to biological incompatibility. PMID- 14562969 TI - Ovarian artery: angiographic appearance, embolization and relevance to uterine fibroid embolization. AB - PURPOSE: To describe the angiographic appearance of the ovarian artery and its main variations that may be relevant to uterine fibroid embolization. METHODS: The flush aortograms of 294 women who had been treated by uterine artery embolization for fibroids were reviewed. Significant arterial supply to the fibroid, and the origin and diameter of identified ovarian arteries were recorded. In patients with additional embolization of the ovarian artery, the follow-up evaluation also included hormonal levels and Doppler imaging of the ovaries. RESULTS: A total of 75 ovarian arteries were identified in 59 women (bilaterally in 16 women and unilaterally in 43 women). All ovarian arteries originated from the aorta below the level of the renal arteries with a characteristic tortuous course. Fifteen women had at least one enlarged ovarian artery supplying the fibroids. Fourteen women (14/15, 93%) presented at least one of the following factors: prior pelvic surgery, tubo-ovarian pathology or large fundal fibroids. CONCLUSION: We advocate the use of flush aortography in women with prior tubo-ovarian pathology or surgery or in cases of large fundal fibroids. In the case of an ovarian artery supply to the fibroids, superselective catheterization and embolization of the ovarian artery should be considered. PMID- 14562970 TI - Neoadjuvant intra-arterial chemotherapy for locally advanced uterine cervical cancer: clinical efficacy and factors influencing response. AB - PURPOSE: To evaluate the effects of neoadjuvant intra-arterial chemotherapy (NAIC) for locally advanced uterine cervical cancer, and to analyze factors influencing the response to the chemotherapy. METHODS: Thirty-four patients with invasive cervical cancer more than 4 cm in diameter were enrolled in this study. NAIC was performed using cisplatin-based regimens. The response was assessed by magnetic resonance imaging (MRI) and examination of surgical specimens. Pretreatment factors involved in the response to NAIC were evaluated and the relationship between the factors and the prognosis was assessed. RESULTS: Clinical response was achieved in 28 (82%) patients. Thirty-one of 49 invasions in the parametrial halves disappeared. Seventeen of 28 lymph node swellings responded to NAIC. Six of the 14 stage III patients became operable. In the 19 surgical cases, pathologically complete responses were found in four. Twenty eight of the 38 parametrial halves were free from cancer. No lymph node metastases were found in eight patients. Initial tumor volume was found to be an independent, significant determining factor of the response to NAIC. Patients with initial tumor volumes less than 80 cm3 had a significantly better estimated 5-year disease-free survival rate compared with those with larger tumors. CONCLUSION: NAIC for locally advanced cervical cancer is useful for preoperative tumor reduction. Tumor volume is a significant determining factor for the response to NAIC. PMID- 14562971 TI - Patency of Wallstents placed at the venous anastomosis of dialysis grafts for salvage of angioplasty-induced rupture. AB - PURPOSE: To retrospectively evaluate the patency of Wallstents placed at the venous anastomosis of polytetraflouroethylene (PTFE) hemodialysis grafts to preserve function following angioplasty-induced rupture. METHODS: The charts of all patients who underwent percutaneous angioplasty of functioning hemodialysis PTFE grafts between September 1997 and September 2001 were retrospectively reviewed. A total of 414 angioplasties were performed. Nine cases of rupture at the venous anastomosis managed with stent placement were identified (7 women, 2 men). Two grafts were loop grafts, seven grafts were straight grafts. All stents placed were Wallstents; six stents were 8 x 40 mm, the remaining three were 8 x 20 mm, 8 x 60 mm and 10 x 42 mm. Average follow-up was 13 months. RESULTS: Technical and clinical success of stent placement was 100%. The primary patency rates (+/- SE) of stents placed at the venous anastomosis were 88% (12%) at 30 days, 63% (17%) at 90 days, 33% (18%) at 180 days and 17% (15%) at 360 days. The secondary patency rates (+/- SE) were 89% (11%) at 90 days, 76% (15%) at 180 days and 69% (23%) (6 stents patent) at 360 days. During follow-up, one graft was removed because of infection, one patient died and another was lost to follow-up. A single minor complication of a puncture site hematoma occurred (11%) with no major complications. CONCLUSIONS: This small retrospective series suggests that Wallstent placement following angioplasty-induced venous anastomotic rupture is effective for preserving dialyzable flow in hemodialysis grafts. Patency is comparable to that of stents placed for reasons other than rupture. PMID- 14562972 TI - Mechanical thrombectomy for early treatment of massive pulmonary embolism. AB - We report our technique and results of percutaneous mechanical thrombectomy in a consecutive series of eight patients with massive PE. We also discuss the possible role of mechanical PE thrombectomy. Eight consecutive patients with acute massive PE, with or without hemodynamic impairment, were treated with mechanical thrombectomy. We used a modified 7-fr hydrolyzer catheter. The treatment was combined with systemic fibrinolysis. From the logistic and technical point we encountered no problems. All patients showed significant improvement while still in the angiography suite. There were no bleeding complications and no other events related to the procedure. Despite the clinical improvement, one patient died shortly after the procedure from cardiac failure. In all patients there was an acute increase in PO2 to normal values. Only a mean of about 50% of all local thrombus could be removed (range 30-80%). The mean PAP pre-intervention decreased only minimally from 42.5 mmHg to 36.3 mmHg post intervention (not significant). In three patients, the PAP continues to remain high at follow-up. The most important feature of mechanical thrombectomy for massive PE is the immediate improvement of the cardiac output, PO2, and clinical situation, overcoming the first critical hours after massive PE. The amount of thrombus reduction seems not to be an important parameter. PMID- 14562973 TI - Clinical outcome following infra-inguinal percutaneous transluminal angioplasty for critical limb ischemia. AB - OBJECTIVE: The aim of this study was to assess the efficacy and durability of infra-inguinal PTA in patients with CLI, in terms of clinical outcome. DESIGN: Retrospective study of 50 consecutive patients with CLI that were exclusively treated by infra-inguinal PTA. METHODS: The indications for intervention were rest pain in seven (14%) patients, non-healing ulcers in 27 (54%), and gangrenous lesions in 16 (32%). Thirty-three (66%) of these patients presented with a single arterial lesion, and the remaining 17 (34%) with multilevel arterial lesions. Kaplan-Meier analysis was used to assess survival, patency, limb-salvage rates, and amputation-free survival. RESULTS: A total of 67 endovascular procedures were performed and 59 (88.1%) of them were considered to be technically successful. The median follow-up period was 12 months (interquartile range: 17 months). The 30-day mortality was 4%, while the cumulative survival rates at 12, 24, and 36 months were 73%, 67%, and 59%, respectively. The cumulative primary patency rates at 12 and 24 months were 63% and 52%, respectively, and remained unchanged thereafter. The estimated secondary patency rate was 72% at 36 months. There was only one below-knee amputation in the patients that were treated exclusively with infra-inguinal PTA. The cumulative amputation-free survival at the same period was estimated at 60%. CONCLUSIONS: Infra-inguinal PTA had a good early and late outcome in this series of patients with a limited life expectancy. These results are comparable to historical results of surgical revascularization in the treatment of CLI. There is need for a randomized study to determine the primary optimal interventional approach for patients with CLI. PMID- 14562974 TI - Management of splenic artery aneurysms and false aneurysms with endovascular treatment in 12 patients. AB - PURPOSE: To assess the endovascular treatment of splenic artery aneurysms and false aneurysms. METHODS: Twelve patients (mean age 59 years, range 47-75 years) with splenic artery aneurysm (n = 10) or false aneurysm (n = 2) were treated. The lesion was asymptomatic in 11 patients; hemobilia was observed in one patient. The lesion was juxta-ostial in one case, located on the intermediate segment of the splenic artery in four, near the splenic hilus in six, and affected the whole length of the artery in one patient. In 10 cases, the maximum lesion diameter was greater than 2 cm; in one case 30% growth of an aneurysm 18 mm in diameter had occurred in 6 months; in the last case, two distal aneurysms were associated (17 and 18 mm in diameter). In one case, stent-grafting was attempted; one detachable balloon occlusion was performed; the 10 other patients were treated with coils. RESULTS: Endovascular treatment was possible in 11 patients (92%) (one failure: stenting attempt). In four cases among 11, the initial treatment was not successful (residual perfusion of aneurysm); surgical treatment was carried out in one case, and a second embolization in two. Thus in nine cases (75%) endovascular treatment was successful: complete and persistent exclusion of the aneurysm but with spleen perfusion persisting at the end of follow-up on CT scans (mean 13 months). An early and transient elevation of pancreatic enzymes was observed in four cases. CONCLUSION: Ultrasound and CT have made the diagnosis of splenic artery aneurysm or false aneurysm more frequent. Endovascular treatment, the morbidity of which is low, is effective and spares the spleen. PMID- 14562975 TI - Anatomy of the portal vein bifurcation: implication for transjugular intrahepatic portal systemic shunts. AB - PURPOSE: The relationship of the portal vein bifurcation to the liver capsule in Asians, which is an important landmark for transjugular intrahepatic portosystemic shunt, has not previously been described. METHODS: The anatomy of the portal vein bifurcation was studied in 70 adult Chinese cadavers; it was characterized as intrahepatic or extrahepatic. The length of the exposed portion of the right and left portal veins was measured when the bifurcation was extrahepatic. RESULTS: The portal vein bifurcation was intrahepatic in 37 cadavers (53%) and extrahepatic in 33 cadavers (47%). The mean length of the right and left extrahepatic portal veins was 0.96 cm and 0.85 cm respectively. Both were less than or equal to 2 cm in 94% of the cadavers with extrahepatic bifurcation. There was no correlation between the presence of cirrhosis and the location of the portal vein bifurcation (p = 1.0). There was no statistically significant difference in liver mass in cadavers with either extrahepatic or intrahepatic bifurcation (p = 0.40). CONCLUSIONS: These findings suggest that for transjugular intrahepatic portosystemic shunt placement, a portal vein puncture 2 cm from the bifurcation will be safe in most cases. PMID- 14562976 TI - Influence of thrombus age on the mechanical thrombectomy efficacy of the amplatz thrombectomy device in vitro. AB - PURPOSE: To determine the efficacy of thrombectomy for the Amplatz Thrombectomy Device (ATD) according to the age of the thrombus. METHODS: Thrombectomy was performed with the 7 Fr or 8 Fr ATD. Five-day-old or 8-day-old thrombi were made from porcine blood by clotting within silicone tubes to be used in a flow model. Emboli sizes, weight, remaining thrombus and activation time were evaluated. RESULTS: The age of the thrombus had no influence on the activation time necessary for complete thrombolysis. A significant difference was found in the overall amount of embolization only with the 7 Fr device although a similar trend was shown with the 8 Fr device. The amount of embolized thrombotic material was higher for both devices if older thrombi (i.e., 8 days old) were processed. The overall impression was that the 8 Fr device showed slightly better results (less remaining thrombus and embolization). CONCLUSION: A tendency toward higher amounts of embolization was observed if older thrombi were used in the model. Therefore the interventional radiologist should be aware of the potentially higher risk of peripheral embolization when performing mechanical thrombectomy (with the ATD) in older thrombotic occlusions, especially since the higher rate of embolization was mainly due to an increased proportion of larger embolic particles (1 mm), which are clinically more significant. PMID- 14562977 TI - First experimental results with a new type of stent: the double-coil device. AB - PURPOSE: To introduce a new stent design and evaluate its technical properties. METHODS: This stent consists of two nitinol wires partially connected to each other. After delivery through a catheter a tube-like helical stent forms within the artery. After experimental tests in flow models regarding mechanical properties, introduction and delivery technique, 15 stents were implanted into iliac, femoral, and carotid arteries of seven dogs. After 3-12 weeks angiographic follow-up stents were explanted for microscopic examination. RESULTS: Stents with expanded diameters of 5-10 mm can be introduced through a 5 Fr catheter with 0.038 inch luminal diameter. Thrombotic vessel occlusion was observed in one iliac artery after incorrect stent placement with diameter mismatch. Fourteen of 15 stents remained patent and revealed minor intimal hyperplasia in the areas of the stent strut connection points as well as some reduction in medial thickness. CONCLUSION: This new stent design has a small introduction diameter which is independent of the expanded diameter. The stent's principal characteristics may serve as a basis for further special developments. PMID- 14562978 TI - CO2 digital subtraction splenoportography with the "skinny" needle: experimental study in a swine model. AB - PURPOSE: To evaluate the safety and the effectiveness of CO2 splenoportography with the "skinny" needle. METHODS: A flexible, 22 gauge needle ("skinny" needle) was introduced into the exteriorized spleens of five pigs. After checking the intrasplenic positioning with CO2 injection, increasing doses of CO2 (10-60 cm3) were injected using a dedicated CO2 injector with digital imaging. The puncture sites were observed during and after CO2 injections, and after removal of the needle. The spleens were then removed for gross and microscopic examination. RESULTS: In all animals digital subtraction CO2 splenoportograms showed the splenic, extra- and intrahepatic portal veins, and the most distal portion of the superior mesenteric vein. No CO2 extravasation occurred in the spleen. There was no significant bleeding from the puncture site after removal of the needle. Gross and microscopic examination revealed no evidence of splenic rupture or intrasplenic hematoma. CONCLUSION: CO2 splenoportography with the "skinny" needle is a safe and simple method of visualizing the portal vein and its branches. Careful appraisals of the clinical usefulness of the method will be needed in various clinical settings. PMID- 14562979 TI - In vitro polyvinylformaldehyde particle compatibility with chemotherapeutic drugs used for chemoembolization therapy. AB - PURPOSE: Because the effects of pirarubicin and carboplatin on the physical structure of particles made from polyvinylformaldehyde are not well known, we describe an experiment to test the in vitro polyvinylformaldehyde particle compatibility with these drugs used for chemoembolization of bone metastases. MATERIALS AND METHODS: Polyvinylformaldehyde particles (Ultra-Drivalon) were mixed in vitro with either pirarubicin or carboplatin as experimental samples, and with distilled water as control samples, and left for 24 h at 37 degrees C. The particles used measured 150-250 microm and 600-1000 microm in diameter. Particle morphology, including appearance, overall shape, and surface characteristics were examined using a microscope equipped with a videocamera. Particle size was measured by granulometry. Qualitative and quantitative variables were analyzed using, respectively, the two-sided Fisher's exact test and the Wilcoxon signed rank test for paired values, with a significance level of 0.05. RESULTS: No broken particles or microscopic degradations in the appearance, overall shape, or surface characteristics of any particles were observed. The particle size distribution was not significantly different between the experimental samples containing pirarubicin or carboplatin and the control sample of particles with diameters in the same range. CONCLUSION: Particles made from polyvinylformaldehyde can be mixed with pirarubicin or carboplatin without any risk of damaging their physical properties. PMID- 14562980 TI - Systemic pulsatile pressure in type II endoleaks after stent grafting of experimental abdominal aortic aneurysms. AB - PURPOSE: To investigate pressure and maximum rate of rise of systolic pressure (peak dP/dt) in completely excluded aneurysms and endoleaks to determine the hemodynamic impact of endoleaks. METHODS: In mongrel dogs (n = 36) experimental aneurysms were created by insertion of a patch (portion of rectus abdominis muscle sheath) into the infrarenal aorta. In group I (n = 18), all aortic branches of the aneurysm were ligated and all aneurysms were completely excluded by stent grafts. Group II (n = 18) consisted of aneurysms with patent aortic side branches that represented sources of endoleaks. One week (n = 12), six weeks (n = 12), and six months (n = 12) after stent grafting, hemodynamic measurements were obtained in thrombosed aneurysms and proved endoleaks. Systemic blood pressure and intraaneurysmal pressure were simultaneously measured and the respective peak dP/dt were computed. RESULTS: At the six-month follow-up, the systolic-pressure ratio (intraaneurysmatic pressure: systemic pressure) was significantly increased in endoleaks compared to non-perfused areas (0.879 +/- 0.042 versus 0.438 +/- 0.176, p < 0.01, group II) or completely excluded aneurysms (0.385 +/- 0.221, group I). Peak dP/dt ratio (intraaneurysmal peak dP/dt: systemic peak dP/dt) was 0.922 +/- 0.154 in endoleaks, compared to 0.084 +/- 0.080 in non-perfused areas (group II, p < 0.01), and was 0.146 +/- 0.121 in completely excluded aneurysms (group I). The diastolic-pressure ratio was also increased in endoleaks compared to non-perfused areas (0.929 +/- 0.088 versus 0.655 +/- 0.231, p < 0.01, group II) or completely excluded aneurysms (0.641 +/- 0.278, group I). In excluded aneurysms, pressure exposure declined as the length of the follow-up period increased. CONCLUSION: Type II endoleaks transmit pulsatile pressure of near systemic level and indicate insufficient treatment result. In contrast, complete endovascular exclusion of aneurysms results in significantly reduced pressure exposure. PMID- 14562981 TI - Helical CT angiography of abdominal aortic aneurysms treated with suprarenal stent grafting. AB - The endovascular repair of abdominal aortic aneurysm (AAA) with stent grafts is rapidly becoming an important alternative to open repair. Suprarenal stent grafting, recently modified from conventional infrarenal stent grafting, is a technique for the purpose of treating patients with inappropriate aneurysm necks. Unlike open repair, the success of endoluminal repair cannot be ascertained by means of direct examination and thus relies on imaging results. The use of conventional angiography for arterial imaging has become less dominant, while helical computed tomography angiography (CTA) has become the imaging modality of choice for both preoperative assessment and postoperative followup after treatment with stent graft implants. There is an increasing likelihood that radiologists will become more and more involved in the procedure of aortic stent grafting and in giving the radiological report on these patients treated with stent grafts. It is necessary for radiologists to be familiar with the imaging findings, including common and uncommon appearances following aortic stent grafting. The purpose of this pictorial essay is to describe and present normal and abnormal imaging appearances following aortic stent grafting based on helical CTA. PMID- 14562982 TI - Digital subtraction fluoroscopy to enhance visualization during uterine fibroid embolization. AB - We describe a simple but underutilized technique for improving visualization during transcatheter embolization using particulate agents. The technique is of distinct utility in uterine fibroid embolization, during which non-target embolization can be of particular clinical significance. PMID- 14562983 TI - Intrasellar latrogenic carotid pseudoaneurysm: endovascular treatment with a polytetrafluoroethylene-covered stent. AB - This case illustrates successful treatment of a large postoperative intrasellar pseudoaneurysm with a polytetrafluoroethylene (PTFE)-covered stent. The advantages and potential disadvantages of this novel method of treatment are discussed. A previously healthy 59-year-old man underwent transsphenoidal operative treatment for hypophyseal macroadenoma, complicated by bleeding. On the 17th postoperative day the patient had profuse arterial bleeding from his nose requiring posterior tamponade. Subsequent angiography of the left internal carotid artery (ICA) revealed a large pseudoaneurysm located intrasellarly, with the orifice in the anteromedial wall in the ophthalmic segment of the ICA. Surgical treatment of the pseudoaneurysm was considered very risky. A 12-mm PTFE covered stent (JoMed, Ulestraten, Netherlands), manually compressed on a PTCA balloon (Maxxum 4.5/13 mm, Boston Scientific, Ireland) was endovascularly delivered covering the orifice of the pseudoaneurysm. Control angiography immediately after the intervention and one year later revealed no flow into the pseudoaneurysm and the ICA was fully patent. In conclusion, deployment of a PTFE covered stent proved to be feasible and successful in the treatment of an intrasellar iatrogenic ICA pseudoaneurysm. Adequate anticoagulative treatment after the procedure is essential to prevent thrombotic complications. PMID- 14562984 TI - Injection of subphrenic saline during radiofrequency ablation to minimize diaphragmatic injury. AB - An 82-year-old man with a history of colorectal cancer presented with metastatic disease to the liver (Couinaud segment 8). We describe the techniques that we employed to successfully perform radiofrequency ablation of a liver metastasis near the dome of the diaphragm utilizing subphrenic infusion of normal saline. The aim of this technique was to prevent thermal injury of the diaphragm. PMID- 14562985 TI - Endovascular mechanical thrombectomy of an occluded superior division branch of the left MCA for acute cardioembolic stroke. AB - Cardiac embolism accounts for a large proportion of ischemic stroke. Revascularization using systemic or intra-arterial thrombolysis is associated with increasing risks of cerebral hemorrhage as time passes from stroke onset. We report successful mechanical thrombectomy from a distal branch of the middle cerebral artery (MCA) using a novel technique. A 72-year old man suffered an acute ischemic stroke from an echocardiographically proven ventricular thrombus due to a recent myocardial infarction. Intraarterial administration of 4 mg rt-PA initiated at 5.7 hours post-ictus failed to recanalize an occluded superior division branch of the left MCA. At 6 hours, symptomatic embolic occlusion persisted. Mechanical extraction of the clot using an Attracter-18 device (Target Therapeutics, Freemont, CA) resulted in immediate recanalization of the MCA branch. Attracter-18 for acute occlusion of MCA branches may be considered in selected patients who fail conventional thrombolysis or are nearing closure of the therapeutic window for use of thrombolytic agents. PMID- 14562986 TI - Utility of magnetic resonance imaging in cardiac venous anatomic variants. AB - The incidence of persistent left superior vena cava (PLSVC) is approximately 0.5% in the general population; however, the coexistent absence of the right SVC has a reported incidence in tertiary centers of 0.1%. The vast majority of reports are limited to pediatric cardiology. Likewise, sinus of Valsalva aneurysm is a rare congenital anomaly, with a reported incidence of 0.1-3.5% of all congenital heart defects. We present a 71-year-old patient undergoing preoperative evaluation for incidental finding of aortic root aneurysm, and found to have all three in coexistence. Suggestive findings were demonstrated on cardiac catheterization and definitive diagnosis was made by magnetic resonance imaging. The use of MRI for the diagnosis of asymptomatic adult congenital heart disease will be reviewed. PMID- 14562987 TI - Ultrasonic-guided percutaneous injection of pancreatic pseudoaneurysm with thrombin. AB - Pancreatic pseudoaneurysm is a relatively uncommon complication of chronic pancreatitis, with an associated high mortality if rupture or hemorrhage occurs. We present a case of pancreatic pseudoaneurysm complicating pancreatitis which was successfully treated by direct percutaneous injection of thrombin into the aneurysmal sac. Follow-up at 8 weeks did not demonstrate recurrence. This case indicates that percutaneous thrombin injection offers effective treatment of visceral arterial pseudoaneurysms. PMID- 14562988 TI - Arterial embolization as a treatment of chronic edema following free cutaneous flap transfer to the lower limbs. AB - Persisting chronic edema following postoperative free flap transfer is a common complication. We report 3 cases of successful treatment by selective arterial embolization of the feeding arterial branches without side effect. One session embolization is advocated as a first line treatment of this condition. PMID- 14562989 TI - The temporal and spatial subnuclear organization of skeletal gene regulatory machinery: integrating multiple levels of transcriptional control. PMID- 14562990 TI - Functional cooperativity between osteoblast transcription factors: evidence for the importance of subnuclear macromolecular complexes? AB - The accompanying article by Drs. Lian and Stein describes current thinking on how genes are organized in the nucleus and suggests that subnuclear localization is critical for the control of gene expression in bone. In particular, it is proposed that a major function of the osteoblast transcription factor, Runx2, is to tether genes that are active in osteoblasts to the nuclear matrix and serve as an organizing center for other nuclear factors which together form osteoblast specific transcriptional units. Although it is still not established that the nuclear matrix localization function of Runx2 is essential for all its biological activities, there is no question that this factor plays a central role in mediating the response of osteoblasts to a variety of signals, as would be expected if Runx2 were involved in organizing the transcriptional apparatus. As will be discussed, Runx2 is required for the response of osteoblasts to other lineage-specific transcription factors and signals initiated by extracellular matrix-integrin binding, growth factors, hormones, and morphogens. We hypothesize that Runx2 transduces this wide range of responses by cycling between active, phosphorylated and a less active, dephosphorylated states which can selectively interact with other nuclear factors to form macromolecular complexes active in transcription. The possible relationship between these complexes and the subnuclear localization of the osteoblast transcriptional apparatus will also be discussed. PMID- 14562991 TI - Association and linkage disequilibrium analyses suggest genetic effects of estrogen receptor alpha and collagen IA1 genes on bone mineral density in Caucasian women. AB - Estrogen receptor alpha (ER alpha) and collagen IA1 (COLIA1) genes have been suggested as possibly implicated in reduced bone mineral density (BMD). The present study investigated the occurrence of association and linkage disequilibrium between radiographic hand BMD and polymorphic alleles of ER alpha and COLIA1 genes, in human pedigrees of a Chuvasha population in Russia. The study sample included 463 members of 113 pedigrees, mostly nuclear families. We performed association and transmission disequilibrium test (TDT) analyses of the combined PvuII and XbaI RFLPs alleles on the same chromosome (haplotype) of the ER alpha gene with BMD Z scores of cancellous or cortical bone in the hand phalanges. The association analyses were performed separately for both genders in the parental generation, i.e., 'fathers' (n = 114; average age 64.2 y) and 'mothers' (n = 122; average age 62.7 y). The Px haplotype was associated significantly with lower BMD Z scores in 'mothers' only. The difference between subjects who carried one or two copies of the Px haplotype and those lacking it was 0.68 Z scores, P = 0.003 and 0.51 Z scores, P = 0.025 for cancellous and cortical bone, respectively. Multiple linear regression model with age, height, weight, and Px haplotype status as predictors explained 26.7% and 28.3% of the total observed variance in BMD with Px haplotype as independent predictor explaining 5.9%; P = 0.002 and 3%; P = 0.028 (cancellous and cortical bone, respectively). Results of t-TDT for triads of two parents and just one of their female offspring (but not male offspring) suggested the existence of linkage disequilibrium between the two loci of Px haplotype and BMD trait (P = 0.047). No association was found between polymorphic alleles of COLIA1 gene and BMD, but 'mothers' with combined genotypes of Px haplotype of ER alpha gene and "s" allele of COLIA1 gene had the lowest mean Z scores (-0.944 and -0.788 for cancellous and cortical bone, respectively). We conclude that the Px haplotype of the ER alpha gene is associated with low BMD values in females, as the phenotype is gender dependent (the association was not observed in males), and the "s" allele of COLIA1 gene in combination with this haplotype contributes to reduced BMD. PMID- 14562992 TI - Suggestive linkage of 2p22-25 and 11q12-13 with low bone mineral density at the lumbar spine in the Irish population. AB - Osteoporosis is a disease characterized by low bone mineral density (BMD) and poor bone quality. Peak bone density is achieved by the third decade of life, after which bone is maintained by a balanced cycle of bone resorption and synthesis. Age-related bone loss occurs as the bone resorption phase outweighs the bone synthesis phase of bone metabolism. Heritability accounts for up to 90% of the variability in BMD. Chromosomal loci including 1p36, 2p22-25, 11q12-13, parathyroid hormone receptor type 1 (PTHR1), interleukin-6 (IL-6), interleukin 1 alpha (IL-1alpha) and type II collagen A1/vitamin D receptor (COL11A1/VDR) have been linked or shown suggestive linkage with BMD in other populations. To determine whether these loci predispose to low BMD in the Irish population, we investigated 24 microsatellite markers at 7 chromosomal loci by linkage studies in 175 Irish families of probands with primary low BMD (T-score < or = -1.5). Nonparametric analysis was performed using the maximum likelihood variance estimation and traditional Haseman-Elston tests on the Mapmaker/Sibs program. Suggestive evidence of linkage was observed with lumbar spine BMD at 2p22-25 (maximum LOD score 2.76) and 11q12-13 (MLS 2.55). One region, 1p36, approached suggestive linkage with femoral neck BMD (MLS 2.17). In addition, seven markers achieved LOD scores >1.0, D2S149, D11S1313, D11S987, D11S1314 including those encompassing the PTHR1 (D3S3559, D3S1289) for lumbar spine BMD and D2S149 for femoral neck BMD. Our data suggest that genes within a these chromosomal regions are contributing to a predisposition to low BMD in the Irish population. PMID- 14562993 TI - Interaction of nutritional calcium and HRT in prevention of postmenopausal bone loss: a prospective study. AB - The aim of this study was to investigate the interactive effects between nutritional calcium (Ca) intake and hormone replacement therapy (HRT) on bone loss. The study population, 937 peri- and postmenopausal women, was selected from a random sample (n = 2025) of the OSTPRE-study cohort (n = 13,100) in Kuopio, Finland. Of them, 545 women had never used HRT and 392 women reported its use during the follow-up period of 6 years. Women were divided in groups according to self-reported daily nutritional Ca intake (mg/day): <648 (1st), 648-927 (2nd), >927 (3rd). Bone mineral density of the lumbar spine and femoral neck was measured with dual X-ray absorptiometry at baseline in 1989-91 and at the 5-year follow-up in 1994-97. According to analysis of variance, there were no statistically significant differences in annual bone loss rate between Ca intake tertiles in HRT never users. In HRT users the annual bone loss at the femoral neck was significantly lower in the third tertile than in the second and first tertiles. In a linear regression model, Ca intake prevented femoral bone loss in HRT users (P < 0.001) but contrast had no effect in never users. At lumbar spine, the corresponding Ca effect was weak (P = 0.063). Adjustment for potentially modifying parameters did not change these effects. In addition, HRT prevented femoral bone loss only among women with the highest Ca intake. At the lumbar spine, the difference between HRT users/non-users was significant in all tertiles but was greater in the second and third tertiles than in the first. In conclusion, nutritional Ca intake may protect HRT users from bone loss and vice versa, low nutritional calcium intake may be a risk factor for non-response to HRT. PMID- 14562994 TI - Influence of weight-bearing exercises on bone acquisition in prepubertal monozygotic female twins: a randomized controlled prospective study. AB - The aim of this study was to determine whether high-impact exercises have an osteogenic effect in 21 prepubertal female monozygotic twins aged 8.7 (SD 0.7) years. Bone mineral density (aBMD), bone mineral content (BMC), bone area, and body composition were derived from DXA. Skeletal maturity was assessed by the Tanner-Whitehouse technique. Anthropometric dimensions (28 dimensions) were measured and also used to derive adiposity and muscularity indices, and information about physical activity was obtained by questionnaire. These measurements were taken before and after the exercise period. The exercise program consisted of high-impact exercises. During 9 months, one girl of each twin pair performed the exercises 3 times a week under supervision of a teacher while their sisters served as control group. At baseline there were no differences between the groups. After 9 months, exercisers (Ex) and controls (Con) had similar increases in height and weight. Significant lower adiposity was observed in the exercise group vs the control group. None of the bone indices differed significantly between the two groups. When the analyses were done for a subgroup of twin pairs (n = 12) who did not participate in high-impact sports during their leisure time, significant differences were obtained for aBMD and BMC of the proximal femur in favor of the exercise group. These results indicate that for prepubertal girls who are not involved in sport activities or who are only involved in low-impact sport activities this exercise program has an osteogenic effect on the proximal femur, but for girls who are already involved in high impact sports this protocol does not have any additional effect on the bone status. PMID- 14562995 TI - Bone turnover responses to changed physical activity. AB - The aim of the study was to compare bone turnover in male soccer players with controls and to follow bone turnover with changes in activity level. Serum osteocalcin (OC), carboxy-terminal propeptide of type I collagen (PICP) and total alkaline phosphatases (tALP) were measured to assess bone formation. Bone resorption was detected by carboxyterminal cross-linked telopeptide of type I collagen (ICTP). Bone turnover of 12 male premier league soccer players (mean age 23 years, range, 17-34) exercising 12 hours/week (range, 8-15) were at the last day of the soccer season compared with 27 age- and gender-matched controls. Bone turnover was followed weekly during a 4-week resting period between two seasons, and a further 10 days following resumption of full training. Data are presented as mean +/- SEM. Both OC (22 +/- 12%) and ICTP (34 +/- 17%) were higher in the players compared with the controls at the end of the season (both P < 0.05, respectively). After 2 weeks of reduced physical activity among the athletes, the PICP levels were 21 +/- 4% (P < 0.05) lower and the ICTP levels 8 +/- 12% higher (P = 0.07) compared with baseline. OC, PICP, and tALP was then no different compared with controls and ICTP was higher than controls (P < 0.001). Ten days within the new season, there was a 23 +/- 5% increase in PICP (P < 0.001) and a 4 +/- 4% decrease in ICTP (P < 0.05) compared with the end of the resting period. In summary, male soccer players have higher bone turnover compared with age- and gender-matched controls. Changes in physical activity level were associated with changes in bone formation and resorption as evaluated by bone markers within weeks, and after 2 weeks rest, ICTP was higher in the athletes than the controls. We conclude that the higher age-related diminution in BMD, previously reported in former soccer players compared with age- and gender-matched controls, may be the result of increased bone resorption, evaluated by ICTP, compared with the controls. PMID- 14562996 TI - Influence of age, sex, and growth variables on phalangeal quantitative ultrasound measures: a study in healthy children and adolescents. AB - Skeletal status by phalangeal quantitative osteosonography (DBM Sonic BP-IGEA) was examined in 1227 healthy children (641 boys and 586 girls) aged 3-16 years. Aims of the study were to evaluate some physical parameters pertaining to the ultrasound transmission crossing the phalanx in a school-age population and to relate these values to age, sex, and growth variables. A correlation was found between AD-SoS (amplitude-dependent speed of sound) and BTT (bone transmission time) and, age, height, weight, and pubertal stage, respectively. No correlation existed between FWA (fast wave amplitude) and SDy (dynamics of the ultrasound signal) and age, height, weight, pubertal stage, and BMI, respectively. AD-SoS increased in boys until 7-8 years of age. Thereafter a plateau was reached up to age 12-13 years, when a rapid increase was observed corresponding to pubertal growth rate acceleration. In girls, AD-SoS increased with age up to 10-11 years with a steeper increase at the time of puberty starting about 2 years earlier than in boys. BTT presented a similar trend. Mean AD-SoS values increased from Tanner pubertal stages 1 to 2 and from stage 3 to 4 in both sexes. Significantly higher mean AD-SoS values in stages 2, 3, and 4 were observed in girls as compared to boys. Mean BTT values increased significantly from stage 1 to 5 in girls and from 1 to 4 in boys. QUS technology showed the ability to assess bone changes in the growing bone. PMID- 14562997 TI - Bone mineral density of the spine and femur in early postmenopausal Turkish women with endemic skeletal fluorosis. AB - The aim of this prospective, comparative study was to investigate the bone mineral density (BMD) changes in a group of early postmenopausal Turkish women with endemic skeletal fluorosis and to study effects of endemic fluorosis on BMD. Bone mineral density of L2-L4 vertebra, femur neck, femur trochanter, and Ward's triangle were measured in 45 female patients with endemic skeletal fluorosis and 41 age-matched controls by dual X-ray absorbtiometry (DXA). The BMD of L2-L4 vertebra and Ward's triangle were higher in the endemic fluorosis group than in the control group (P < 0.001). Patients with endemic fluorosis had higher femur neck and femur trochanter BMDs than did controls (P < 0.01 and P < 0.05, respectively). There was a positive correlation between serum fluoride content and BMD at the spine (r = 0.345, P = 0.001), femoral neck (r = 0.274, P = 0.011), Ward's triangle (r = 0.295, P = 0.006), and trochanter (r = 0.217, P = 0.045). In conclusion, higher bone mineral density levels were seen in early postmenopausal women with endemic skeletal fluorosis. BMD measurement is a tool in the diagnosis and management of this preventable crippling disease. PMID- 14562998 TI - Monitoring sweat calcium using skin patches. AB - The purpose of this study was to determine whether a simple noninvasive sweat collection method using skin patches would be useful in monitoring sweat Ca and to determine changes in dermal Ca loss during a bed rest study testing a resistive exercise countermeasure. The study showed that the technique was highly reproducible: the mean intra-subject variation approached zero and the inter individual variability (%CV) varied from 18% to 32% for the three anatomical regions (arm, chest, and back) tested. There was less than 10% difference in sweat Ca excretion from different skin regions within the same individual at a given time point. A calculated estimate of total body sweat excretion for 12 bed rest subjects was 35 +/- 4 mg/day (mean +/- SE), close to published whole body measurements. Bed rest testing showed no significant differences with or without exercise when conducted in a temperature-controlled environment. We conclude that the skin patch technique is useful for monitoring changes in sweat Ca. PMID- 14562999 TI - Microstructural classification of resorption lacunae and perforations in human proximal femora. AB - Resorption lacunae and perforation types mirror resorptional activity of osteoclasts and are important for the integrity of bone architecture. In the present study, distinctive microstructural features of cancellous bone dissected from femoral heads of 28 autopsy subjects (14 females and 14 males) were defined and evaluated by light microscopic (LM) histology and scanning electron microscopy (SEM). The work differentiates two types of resorption lacunae in trabecular bone: the longitudinally extended resorption lacunae (LER) and the reticulate patch resorption lacunae (RPR). Further, two types of perforations are distinguished: the lacunar perforation (LP) and the tunneling perforation (TP), which are differentiated from potential blood vessel canals (BC). Evidence is presented that the spatial distribution on rods or plates is highly correlated with the resorption type. The RPR type was more frequently seen and was primarily localized at the nodes of rods, in the middle regions of rods, and in the center region of plates as compared to the LER type. The presented evaluation scheme of resorption and perforation types could prove useful in future studies for systematically investigating potential microstructural changes associated with disturbed bone turnover. PMID- 14563000 TI - Effects of SP500263, a novel selective estrogen receptor modulator, on bone, uterus, and serum cholesterol in the ovariectomized rat. AB - We describe here the activity of a novel selective estrogen receptor modulator, SP500263. When given to adult ovariectomized (OVX) rats for 28 days at doses of 0.3, 1, or 3 mg/kg/day, we found that SP500263 partially protected against OVX induced loss of bone mineral content in the distal ends of femurs and in the whole bone. SP500263 also antagonized the OVX-induced increase in body weight. However, unlike 17beta-estradiol, SP500263 at efficacious doses did not prevent the OVX-induced loss in uterine wet weight. A small but significant effect on uterine wet weight was noted with raloxifene dosed at 1 mg/kg. As expected, SP500263 but not raloxifene acted as an estrogen antagonist on the uterus in adult rats when administered for 7 days at 30 mg/kg/day. Finally, SP500263 had no statistically significant effects on total serum cholesterol and serum triglycerides in OVX rats treated for 28 days. Raloxifene had no significant effects on body weight, bone mineral content, and serum cholesterol or triglycerides in the OVX-rat model. In summary, SP500263 is a new orally active SERM that acts in rats as an estrogen agonist on bone without causing uterine stimulatory effects. PMID- 14563001 TI - Composition of biomineral organic matrices with special emphasis on turbot (Psetta maxima) otolith and endolymph. AB - The soluble organic matrix (OM) of various biominerals (red coral skeleton, oyster shell, sea urchin test, turbot otolith, chicken eggshell) was extracted after demineralization with acetic acid. The protein content of the OM varies strongly from 0.02 to 1.6 microg/mg biomineral whereas proteoglycans present less variations (from 0.7 to 1.4 microg/mg biomineral). Electrophoresis of biominerals OM shows differences in their protein pattern although several bands are present in all matrices. OM of all biominerals shows carbonic anhydrase activity but no activity was detectable in the endolymph. OM of all biominerals also displays an anticalcifying activity. After separation of the OM extracts by chloroform methanol, 80% of the anticalcifying activity was found in the methanol phase except in the urchin test. After OM precipitation with trichloracetic acid, 70% of the activities was found in the supernatants. Partial biochemical characterization suggests that the anticalcifying factor is a polyanionic and water-soluble molecule, which could be proteoglycans. The endolymph surrounding the otolith also displays an anticalcifying activity although its inhibitous activity was 50 times lower than that of the otolith OM. However, the anticalcifying activity of the endolymph is assumed by a proteic structure (80% activity precipitated with TCA treatment). Our results suggest that both carbonic anhydrase and anticalcifying activities are widespread and play a significant role in the regulation of biomineral formation. Results are discussed in relation to the calcification process that takes place at the fluid-mineral interface. PMID- 14563002 TI - Quantitative evaluation of the biocompatible and osteogenic properties of a range of biphasic calcium phosphate (BCP) granules using primary cultures of human osteoblasts and monocytes. AB - A range of 50% porous gamma-sterilized biphasic calcium phosphate (BCP) granules, (20, 50, 80, and 100% tricalcium phosphate, TCP) were classified into two distinct size ranges, small 2-4 mm in diameter and large 4-6 mm in diameter, and their potential as bone graft extender materials was assessed in vitro using culture systems of primary-derived peripheral human blood monocytes and human osteoblasts isolated from bone. The effect of the in vitro culture conditions was evaluated prior to the introduction of the test substrates. The cellular response was assessed via quantification of viable cell adhesion to the materials, lactate dehydrogenase (LDH), the production and release of interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), and prostaglandin E2 (PGE2). The higher content TCP materials, 80% and 100% TCP, had a detrimental effect on viable cell adhesion after day 1, which was not related to calcium release from the granules within the local environment. TCP granules (20% and 50%) initiated a controlled level of inflammatory response that sustained and promoted viable macrophage adhesion throughout the test period, The percentage of TCP within the BCP granules was a governing factor in determining the cellular response. PMID- 14563004 TI - Volumes from which calcium and phosphorus X-rays arise in electron probe emission microanalysis of bone: Monte Carlo simulation. AB - Monte Carlo simulations of trajectories for electrons with initial energy of 10 keV through 30 keV were used to map the 3D location of characteristic x-ray photon production for the elements C, P, and Ca until the electrons either escaped as backscattered electrons (BSE) or had insignificant energy. The x-ray production volumes for phosphorus slightly exceed those for calcium, but both greatly exceed the volume through which BSE travel prior to leaving the sample. The x-ray volumes are roughly hemispherical in shape, and the oblate spheroid from which BSE derive occupies only the upper third to half the volume of x-ray generation. Energy-dispersive x-ray emission microanalysis (EDX) may not be secure as a method for the quantitation of BSE images of bone in the scanning electron microscope (SEM). Ca:P elemental ratios from EDX analyses may also be imperfect. PMID- 14563003 TI - Canine cancellous bone microarchitecture after one year of high-dose bisphosphonates. AB - We examined the effects of one-year high-dose bisphosphonates (risedronate 0.5 mg/kg/day or alendronate 1.0 mg/kg/day) on the three-dimensional (3-D) microstructural and mechanical properties of canine cancellous bone. A high resolution micro-CT scanner was used to scan cubic specimens produced from the first lumbar vertebrae. Microstructural properties of the specimens were calculated directly from the 3-D datasets and the mechanical properties of the specimens were determined. Our data demonstrate significant microarchitectural changes in the bisphosphonate-treated cancellous bone that was typically plate like, denser, with thicker and more trabeculae compared with those of the controls. Consistent with architectural changes, the Young's moduli of cancellous bone increased in all three directions with the greatest increase in primary axial loading (cephalo-caudal) direction after treatment. Our results suggest a bone remodeling-adaptation mechanism stimulated by bisphosphonates that increases bone volume fraction, thickens trabeculae, changes trabeculae towards more plate like, and increases mechanical properties. The secondary degree of anisotropy contributed significantly to the explained variance in bone strength, and the primary or tertiary degree of anisotropy improved the explanation of variances for Young's moduli, i.e., 79% of strength variances or 74-83% of modulus variances could be explained by the combined anisotropy and bone volume fraction. These significant improvements of cancellous bone architecture provide a rationale for the clinical observation that fracture risk decreased by 50% in the first year of bisphosphonate therapy with only a 5% increase in bone mineral density. We conclude that bisphosphonates enhance mechanical properties and reduce fracture risk by improving architectural anisotropy of cancellous bone 3-D microarchitecture. PMID- 14563005 TI - Proximal tibial fractures: current treatment, results, and problems. AB - Fractures of the proximal tibia can present unique treatment challenges. Reduction and stability are dependent on control of the proximal fragment. Soft tissue compromise can present as a component of the injury, or can result from surgical dissection. Treatment protocols aimed at addressing these issues have included closed treatment, external fixation, intramedullary nailing, and plating. The issues of accurate reduction, proximal fragment stabilization, and soft tissue protection are discussed with an emphasis on the individual challenges of each of the treatment modalities. PMID- 14563006 TI - LISS PLT: design, mechanical and biomechanical characteristics. AB - Following the development of the Less Invasive Sabilization System for Distal Femur a similar system for proximal tibia fractures (LISS PLT) was designed. Anatomical studies were carried out to define the shape of the plate and the position and orientation of the screws. Standard mechanical tests were performed to ensure that the LISS PLT fixation provides similar fatigue resistance to that of conventional plates. Finally, cadaver tibia pairs were used to compare the biomechanical performance of the new device to that of bilateral plating. An unstable intra-articular proximal tibia fracture model was used. The medial condyle of the tibias were submitted to loading cycles with increasing load levels and the vertical subsidence of the medial condyle during the loading cycles was monitored. Comparable stability against secondary loss of reduction was observed for the LISS PLT and the bilateral plating constructs. PMID- 14563007 TI - Less Invasive Stabilization System (LISS) for fractures of the proximal tibia: indications, surgical technique and preliminary results of the UMC Clinical Trial. AB - The surgical treatment of proximal tibia fractures, with or without intraarticular involvement, is associated with well-described patterns of failure and significant complication rates. Recent surgical advances allow for a minimally invasive approach to such injuries, which may improve healing times, increase union rates, and decrease complication rates. The Less Invasive Stabilization System (LISS) for proximal tibia fractures employs a laterally based fixed angled implant, allowing for the placement of locking screws proximal and distal to the fracture, which may be placed percutaneously. This manuscript presents the surgical technique and indications of the Tibia LISS. A preview of the early clinical results of the treatment of bicondylar tibial plateau fractures and proximal tibia diaphysis will also be presented. PMID- 14563008 TI - Stabilization of proximal tibial fractures with the LIS-System: early clinical experience in Berlin. AB - The Proximal Tibia Less Invasive Stabilization System (LISS PLT) is an internal fixator for the treatment of proximal tibial fractures according to the principles of "Minimally invasive surgery". From July 1998 to August 2000 22 fractures were treated in our clinic with the new Proximal Tibia LISS and the prospective course of healing was documented. The period of follow-up was 12 months. The inclusion criteria were defined as proximal tibial shaft fractures and intraarticular proximal tibial fractures of all degrees of severity. A total of 15 proximal medial and lateral tibial plateau fractures (AO 41) and 7 metaphyseal fractures were treated (AO 42), seven of these fractures presented with open soft tissue damage. The average age of the patients treated was 42 years. With a follow-up rate of 91% (2 patients lost to follow-up), definite consolidation of the fracture was seen in 19/20 cases. In one patient, the fracture had only been partially bridged and secondary bone grafting was performed. Radiologically, there was one case of a varus malalignment of 6 degrees, in two further cases there were valgus malalignments of 5 degrees and 7 degrees at the time of surgery. The other cases all healed in correct alignment. In one case, the implant became loose on the distal shaft and was stabilized again using bicortical screws. In a case with type IIIB soft tissue damage, a soft tissue infection became manifest, but healed uneventfully after a revision operation with the implant in situ. PMID- 14563010 TI - Compartment pressures after submuscular fixation of proximal tibia fractures. AB - INTRODUCTION: Tibial fractures are known to be associated with elevated compartment pressures, which may potentially result in a compartment syndrome. The Tibial Less Invasive Stabilization System (LISS PLT) is an internal fixator device that is designed for fixation of proximal tibia fractures. The technique includes submuscular placement of the implant along the lateral border of the tibia. Such placement of the Tibia LISS into the anterior leg compartment could increase compartment pressures, and therefore increase the risk of compartment syndrome. The objective of this prospective study is to determine whether submuscular placement of a tibia LISS fixator increases compartment pressures and what is the incidence of compartment syndrome. MATERIALS AND METHODS: Twenty eight consecutive proximal tibia fractures were fixed using the Tibia LISS (LISS PLT). Compartment pressures in all four leg compartments were measured pre- and postoperatively. Diastolic pressures were recorded and perfusion pressures calculated. RESULTS: There were no statistically significant differences between pre- and postoperative compartment pressures. One patient out of twenty-eight developed an impending compartment syndrome after placement of the LISS implant. CONCLUSIONS: Submuscular fixation of the tibia with the Less Invasive Stabilization System does not cause a significant rise in leg compartment pressures, nor is it associated with an increased incidence of compartment syndrome. PMID- 14563009 TI - Fracture stabilization of proximal tibial fractures with the proximal tibial LISS: early experience in Birmingham, Alabama (USA). AB - This study is a prospective evaluation of the Less Invasive Stabilization System (LTSS) for the treatment of high-energy tibial plateau and proximal tibial fractures treated between November, 1998 and June, 2000. Thirty-two patients sustained thirty-five acute fractures of the tibial plateau (25) or proximal tibia (10). These patients were injured primarily in blunt trauma accidents, with eighteen having multiple fractures, fifteen having ipsilateral extremity fractures, and eleven having major knee ligament injuries. Seventeen patients had open fractures. Thirty-four patients healed their fractures, with one developing a nonunion. Two patients developed infections, both following Type III open fractures. Final range of motion averaged 2 to 116 degrees. Alignment was well maintained, with no patient losing the alignment that was obtained in the operating room. The tibial LISS system worked well at stabilizing difficult fractures of the tibial plateau and proximal tibia with a low incidence of complications in this preliminary study with short-term follow-up. PMID- 14563011 TI - Silicone gel breast implants and connective tissue disease--a comprehensive review. AB - Silicone breast implants have been in use for breast reconstruction and breast augmentation for a long time. In the late 80's anecdotal reports describing a possible association between silicone-gel filled breast implants, and autoimmune diseases was accumulating. Due to the growing concern about the safety of silicone-gel implants, the Food and Drug administration (FDA) restricted their use to participants in controlled clinical trials, including women having reconstructive surgery. However, in recent years, large meta-analyses were unable to establish an association between silicone-gel breast implants and autoimmune disease. We review the current literature, emphasizing the most important epidemiologic studies. PMID- 14563012 TI - Mechanisms of immunological tolerance loss versus erythrocyte self-antigens and autoimmune hemolytic anemia. AB - Recent studies on animal and human autoimmune hemolytic anemia (AIHA) suggest that immunological tolerance loss toward red blood cells (RBC) self-antigens may be originate by different, non-mutually exclusive, mechanisms. According to now available data the identified mechanisms may be: ignorance against RBC self antigens; molecular mimicry; polyclonal T and/or B cells activation; errors in central or peripheral tolerance; immunoregulatory disorders including cytokine network alteration. In some patients with AIHA, stimulation of PMBC by synthetic Rh peptides indicate that ignorant T and/or B cell clones may recognize cryptic RBC self-antigens. AIHA associated with bacterial or viral infections seems to be produced by polyclonal T and/or B cells activation against foreign antigens which mimic protein or carbohydrate epitopes on RBC. Polyclonal activation of host B cell clones by donor alloreactive T cells causes the AIHA in chronic GVHD. As the tolerance loss is concerned, experiments on mouse lines expressing a transgene with autoantibody activity against murine RBC have shown that non-deleted peripheral B cell clones may produce RBC autoantibodies. In humans a genetic defect of Fas/FasL autoreactive lymphocytes apoptosis may be associated to AIHA. Immunoregulatory disorders due to depletion of CD4+ CD25+ T cells or Th1/Th2 cytokines imbalance may induce autoimmune diseases. In mice AIHA may be induced or improved by cytokines or anticytokine antibodies administration. In NZB/W mice and human AIHA there is an increased production of Th2 cytokines as IL4 and IL10 but INF-gamma reduced production. In addition in human AIHA has been shown a downregulation of IL12 and therefore, an IL10/IL12 immunoregulatory circuit imbalance which might facilitate the RBC autoantibodies production. PMID- 14563013 TI - Single immunization of newborn mice with heterologous type-II collagen induces arthritic disease. AB - Collagen-induced arthritis (CIA) is a widely accepted model of autoimmune disease with significant similarities to rheumatoid arthritis in humans. CIA is provoked in susceptible strains upon immunization of adult mice with native type-II collagen in complete Freund's adjuvant (CFA). Neonatal exposure to antigen is supposed to result in T cell clone deletion and induction of tolerance. Here we report that the neonatal injection of bovine type-II collagen (bCII) to ICR (CD 2) mice triggers the development of autoimmune chronic joint inflammation. Compared with standard CIA significant joint swelling was not observed and anti collagen antibodies were not detected if the second challenge with the antigen was not supplied. Histopathologic examination of the joints showed cell infiltration, synovial hyperplasia and at the later period bone destruction. Mice immunized as neonates expressed Ag-specific proliferative response and delayed type hypersensitivity (DTH) reaction to bCII. PMID- 14563014 TI - Cytokine sensitivity of Langerhans' islets of diabetes-prone BB/OK rats under hypoglycemic conditions. AB - Islets of Langerhans isolated from diabetes-prone BB/OK rats were exposed to interleukin-1beta (IL-1beta) or to a combination of tumor necrosis factor-alpha (TNF-alpha) plus interferon-gamma (IFN-gamma) under hypoglycemia at glucose concentrations of 2.2 and 3.2 mmol/l or in the presence of stimulatory conditions at 6.0 and 11 mmol/l glucose. For estimating cytokine effects the islets were functionally assayed by measurement of glucose stimulated insulin secretion. Pancreatic islets exposed for 24 h to IL-1beta at a glucose concentration of 6.0 mmol/l exhibited a reduced insulin secretion following a 48h recovery period compared to islets which were cytokine treated at 2.2 or 3.2mmol/l glucose, respectively. Islets pre-exposed for 24h to TNF-alpha plus IFN-gamma at 2.2, 3.2 or 6.0 mmol/l glucose displayed no alterations of insulin secretion following a 48 h regeneration. A temporary (3 h) influence of IL-1beta under hyperglycemic conditions at 11 mmol/l glucose caused a reduction of the subsequent insulin secretion of Langerhans' islets prior incubated for 24 h at 6.0 mmol/l glucose without cytokines, but not of islets precultured at 2.2 mmol/l glucose. In contrast, a 3 h treatment with TNF-alpha plus IFN-gamma at 11 mmol/l glucose did not affect insulin secretion of islets prior held at 6.0 mmol/l glucose, whereas a transient exposure for 6h to IL-1beta as well as TNF-alpha plus IFN-gamma under similar conditions diminished insulin secretion of islets preincubated at 2.2 or 6.0 mmol/l glucose. In conclusion, hypoglycemia reduces the sensitivity of BB/OK rat islets to IL-1beta, whereas a slight elevation of glucose concentration to 6.0 mmol/l increases again their vulnerability. TNF-alpha plus IFN-gamma at concentrations capable to decrease insulin secretion of islets during hyperglycemia do not affect the insulin output in a range between 2.2 and 6.0 mmol/l glucose. During glucose stimulation at 11 mmol/l islets' insulin secretory machinery is protected from IL-1beta as well as TNF-alpha plus IFN-gamma for 3 h by a preceding 24 h hypoglycemia, but its vulnerability is restored within additional 3 h. PMID- 14563015 TI - Biochemical and immuno-pathological aspects of tissue transglutaminase in coeliac disease. AB - Tissue transglutaminase (tTg) has been identified as the major autoantigen in coeliac disease (CD). ELISA methods have been developed for measuring the autoantibody. There are divergent reports on the effects of calcium on the antibody binding to tTg. Furthermore, zinc is a potent inhibitor of tTg. To better understand the role of transglutaminase in CD, we have studied the stability of commercial tTG, the effect of CD serum on tTg-activity and the effects of calcium and zinc on the antibody binding. The inclusion of calcium during the coating of the ELISA plates significantly increases the binding of the antibody, while zinc at physiological concentrations inhibits the binding. Moreover, our results show that commercial guinea pig liver Tg treated with calcium contains at least four major antigenic molecules and is a labile enzyme, which is degraded rapidly by contaminating proteases. Human serum contains anti proteases that protect the enzyme. Probably, the labile character of commercial tTG explains the divergent reports on the effects of calcium on antibody binding. Finally, antibodies in serum from a CD patient do not seem to inhibit tTg activity. Hypothetically, low, intestinal Zn2+ -levels facilitate Ca2+-activation of tTg, which deamidates gliadin. A complex between tTg and modified gliadin forms the antigen and triggers the immune reaction leading to manifest CD. Hypozincaemia secondary to villous atrophy aggravates the induced disease. PMID- 14563017 TI - Structure and function of CD72 in the non-obese diabetic (NOD) mouse. AB - Type I or insulin dependent diabetes mellitus develops in the non-obese diabetic (NOD) mouse as a consequence of T cell mediated autoimmune attack on pancreatic beta cells. B lymphocytes are required for disease progression in NOD and loss of tolerance in the B cell compartment is one of the earliest manifestation of the autoimmune process. To understand how the fate and function of B lymphocytes may be regulated in the context of an organ specific autoimmune disease, the B cell co-receptor CD72 (Lyb-2) was examined in NOD mice. Mab that recognize a,b, and d alleles of CD72 reacted poorly with NOD B cells while western blots of B cell extracts show that CD72 is abundant in NOD B cells. Nucleotide sequencing of CD72 cDNA confirms that an uncommon allele, CD72c, is expressed in NOD. Functional studies using monoclonal antibodies indicate that the CD72c allele of NOD can serve as a positive regulator of B cell responses both as a single signal and in synergy with BCR or IL-4 stimulation. Since CD72c differs principally in the extra cellular or ligand binding portion of the molecule, interactions with its natural ligand in vivo may contribute to functional differences in mouse strains that express this allele. NOD and lupus prone strains share the CD72c allele and its functions may contribute to overlapping features of organ specific and systemic autoimmune disorders. PMID- 14563016 TI - Islet autoantibodies in cord blood from patients who developed type 1 diabetes mellitus at 15-30 years of age. AB - Islet cell autoantibodies are early markers for type 1 diabetes. The aim of this study was to determine whether islet autoantibodies were present at birth in young adults who developed type 1 diabetes at 15-30 years of age. Cord blood sera from 30 patients who developed type 1 diabetes between 15 and 25 years of age and sera from 320 randomly selected control children were tested for islet cell antibodies (ICA) by indirect immunofluorescence and autoantibodies against the 65 kD isoform of glutamic acid decarboxylase (GADA), islet cell antigen-2 (IA-2A) and insulin (IAA) by radiobinding assays. The young adults who developed type 1 diabetes did not differ from controls in the cord blood prevalence of any of the four islet autoantibodies. This is in contrast to our previous findings that children who developed type 1 diabetes below 15 years of age had an increased prevalence of cord blood islet autoantibodies. Our present data suggest that, in contrast to children, pre- and perinatal risk factors are less likely to be involved in the development of type 1 diabetes in young adults. PMID- 14563018 TI - Lrmp and Bcat1 are candidates for the type I diabetes susceptibility locus Idd6. AB - Three type 1 diabetes associated regions on distal mouse chromosome 6 have recently been defined by the construction and analysis of a series of congenic strains, carrying C3H/HeJ genomic material on a NOD/Lt genetic background. Whilst NOD/Lt alleles at the most distal locus Idd6 confer susceptibility, C3H/HeJ alleles confer resistance to diabetes. Idd6 overlaps with a locus controlling low rates of proliferation in immature NOD-thymocytes, suggesting that Idd6 could be controlling diabetes development through an effect on T cell proliferation rates. Candidates for Idd6 therefore include genes, which are implicated in the immune system and/or in the control of cell proliferation rates, such as Lrmp (Jaw1), Bcat1 and Kras2 that map to the Idd6 candidate region. In the present study, we have undertaken an expression and mutational analysis of all three genes. A surprisingly large number of polymorphisms and amino acid changes were identified in both Lrmp and Bcat1 indicating that they are candidates for Idd6. The two genes are located within a genomic interval of about 3 Mb that contains a large number of single nucleotide polymorphisms (SNP) and which has possibly been derived from distinct ancestral haplotypes in the C3H/HeJ and NOD/Lt strains. PMID- 14563020 TI - A brief introduction to unified test theory. AB - This article introduces McDonald's unified treatment of test theory, which merges the major contributions of Spearman (True Score Theory and Common Factor Theory) with aspects of Item Response Theory. The fundamentals are first presented. followed by elaboration of selected aspects of the treatment. An SAS program is given that estimates relevant parameters. PMID- 14563019 TI - Presence of B7-2+ dendritic cells and expression of Th1 cytokines in the early development of sialodacryoadenitis in the IqI/Jic mouse model of primary Sjorgren's syndrome. AB - Subpopulations of infiltrating lymphocytes, professional antigen-presenting cells (APCs), and Th1/Th2 cytokines that could initiate an autoimmune sialodacryoadenitis were studied in the IQI/Jic mouse model of primary Sjogren's syndrome. Although lymphocytic infiltrations were first seen in submandibular glands (SMGs) of females and in lacrimal glands (LGs) of males at 8 weeks of age, clusters of MHC class II+, CD11c+, B7-2+ dendritic cells (DCs) were already localized in these tissues at 4 weeks. At 8 weeks, the infiltrating lymphocytes consisted of almost equal numbers of B cells and CD4+ T cells. In the inflammatory foci, MHC class II+, CD11c+, B7-2+ DCs formed network-like structures. Duct cells in the lesions showed immunoreactivities for MHC class II and ALCAM (a costimulatory adhesion molecule). IL-12 and IFN-gamma transcripts were detected by RT-PCR in SMGs of females and in LGs of males at 8-12 weeks. These results suggest that the clustered DCs might play an important role in the initiation of the adenitis, and further suggest that the DCs and epithelial cells may participate in the activation of CD4+ T cells. It is also likely that Th1 cytokines mediate the functional interactions between the APCs and CD4+ T cells in the early lesions. PMID- 14563021 TI - Associations among job satisfaction, optimism, pessimism, and psychosomatic symptoms for employees in the government sector in Kuwait. AB - The present study examined associations among job satisfaction and optimism, pessimism, and psychosomatic symptoms in a sample of 718 Kuwaiti employees (350 men and 368 women). Men scored significantly higher than women on both the scales of Job Satisfaction and Optimism, while women scored significantly higher than men on both scales for Pessimism and Psychosomatic Symptoms. Job satisfaction scores correlated significantly and positively with Optimism scores and negatively with Pessimism scores. There was also a significant negative correlation between scores on Optimism and Pessimism, and a significant negative correlation between scores on Optimism and Psychosomatic symptoms. Results were discussed within the Kuwaiti context. PMID- 14563022 TI - Development of the overt-covert aggression inventory. AB - The expression of anger in Japanese people is different from that of other races. We developed a new brief inventory, the Overt-Covert Aggression Inventory, to assess aggressive behavior ofJapanese people by focusing on their uniqueness and examined its reliability and validity. This inventory, the Center for Epidemiological Studies Depression scale, the Japanese version of the Buss-Perry Aggression Questionnaire, and the Picture-Frustration Study were administered to 3,104 men and 316 women in a factory. Internal consistency, test-retest reliability, concurrent validity, and construct validity of the scale were examined. We confirmed that the Overt-Covert Aggression Inventory has adequate reliability and sufficient concurrent validity, however, further studies of the construct validity and discriminant validity are required. PMID- 14563023 TI - Scores on the eysenck personality questionnaire for a sample of children and adolescents receiving psychological treatment in Puerto Rico. AB - Personality questionnaire scores obtained by children and adolescents (n = 28) receiving psychological treatment at a health facility in Humacao, Puerto Rico were examined. The scores were compared with those of regular school children of the same age, of Gurabo, Puerto Rico, who were not in treatment (n = 30). The children in treatment obtained higher scores on Psychoticism, lower scores on Extraversion, and similar scores on Neuroticism and Dissimulation by comparison with regular students. PMID- 14563024 TI - Effect of backchannel utterances on facilitating idea-generation in Japanese think-aloud tasks. AB - The relation between backchannel utterance and idea-generation has hardly been studied. Based on preliminary investigations, we formulated a hypothesis that a listener's backchannel utterances facilitate a speaker's idea-generation. This study experimentally manipulated the frequency of backchannel utterances by listeners during speakers' idea-generation for think-aloud tasks. 16 Japanese female undergraduates participated. Analysis indicated that frequent backchannel utterances increased not only the number of ideas generated but also the speaking time for the tasks. PMID- 14563025 TI - Sex differences among five sources of motivation in the motivation sources inventory: preliminary findings. AB - Paper reports sex differences among five sources of motivation for a sample of 208 undergraduate students who completed the Motivation Sources Inventory. t tests yielded few statistically significant sex differences for the five sources of motivation; however, Instrumental Motivation was higher among men than women, while the remaining four motives showed no sex difference. Implications and research opportunities are discussed. PMID- 14563026 TI - Validity and reliability of the general well-being schedule with northern plains American Indians diagnosed with type 2 diabetes mellitus. AB - The General Well-being Schedule is a brief indicator of subjective feelings of psychological well-being and distress. It is easy to administer, reliable, and valid, although its validity with American Indians has not been established. This study then assessed reliability, validity, and factor structure for a sample of 88 diabetic American Indians, who sought care for diabetes at an Indian Health Service hospital. Cronbach alpha was .89. A factor analysis indicated four dimensions. Adequate concurrent and divergent validity were noted in association with scores on the Beck Depression Inventory-Second Edition, the depression scale on the Symptom Checklist-90-Revised, and Family-Adaptation, Partnership, Growth, Affection, & Resolve. These results suggest that the General Well-being Schedule is a reliable and valid measure of general well-being for this population of American Indians. PMID- 14563027 TI - Anger expression: parental and cognitive factors. AB - The associations of parental moral disengagement, guilt, prosocial behavior, and anger, with their children's maladaptive anger was examined. 98 college undergraduate students and their parents participated. Both students and parents completed the Anger Response Inventory, the Mechanism of Moral Disengagement Scale, the Texas Social Behavior Inventory, the Fear of Punishment Scale, and the Need for Reparation Scale. A multiple regression analysis was performed to assess the students' variables which predicted maladaptive anger. Only moral disengagement was a predictor of the students' maladaptive anger. Subsequent multiple regression analyses were used to examine whether parental variables predict students' anger. Fathers' maladaptive anger, and prosocial skills were significantly related to students' maladaptive anger. Maternal variables produced an increase in the multiple R similar to the fathers', but none of the individual measures were significantly associated with the students' maladaptive anger. PMID- 14563028 TI - Wais-III matrix reasoning: instruction effects on task perception and performance. AB - Instructions for the Matrix Reasoning subtest of the WAIS-III do not communicate to examinees that the subtest is untimed. The present study examined what percentage of participants (34 women, 26 men, M age = 20.1; 55 Caucasian, 3 African American, 2 Hispanic) made the assumption that Matrix Reasoning was timed, and its effect on examinees' scores. 55% of participants receiving standard instructions retrospectively reported assuming the subtest was timed, and those who did not assume Matrix Reasoning was timed scored significantly higher than participants who did. Participants receiving additional instructions that clarified the untimed nature of Matrix Reasoning scored significantly higher than those receiving standard instructions who believed the subtest was timed. PMID- 14563029 TI - An approach for practice administration and interpretation of the MMPI-2. AB - This paper discusses one teaching approach for practice administration and interpretation of the MMPI-2 in a graduate course on personality assessment. After graduate students practice scoring and interpreting the MMPI-2 inventory completed with specific response sets and practice interpreting MMPI-2 profiles of cases prepared for teaching and practice in interpretation, students practice administering and interpreting the MMPI-2 for volunteer test-takers who complete the inventory with a simulated stress response set. The simulated stress response set approach may be helpful in providing skill-building practice in the early phases of learning the MMPI-2 with volunteer test-takers, while minimizing some of the ethical concerns raised regarding actual administrations with nonclient volunteers in the early phases of training in assessment. PMID- 14563030 TI - Sensitivity to disgust and perceptions of natural bodies of water and watercraft activities. AB - A written 7-item self-report scale on sensitivity to disgust and participation in watercraft activities was administered to 450 seasonal park employees. Correlations indicate that nonparticipation in seven different watercraft sports was weakly related with reactions of disgust to contact with natural bodies of water (r(pbis) = -.12 to -.32, p < .01). Negative affective reactions to contact with natural bodies of water may contribute to panic reactions in watercraft accidents. PMID- 14563031 TI - Mental health and vitality among Canadian women with physical disabilities. AB - This study investigated scores for mental health and vitality in a large community-based sample of women with physical disabilities. The scores from two subscales of the SF-36 were collected from 1,096 women with physical disabilities through a mailed survey regarding health and well-being. These scores were compared to normative data using t tests. The mean scores of the vitality subscale were significantly lower than that of the normed sample when analyzed by age groups. The mental health scores were significantly lower as well, except for one age group (65-74 yr.). These results suggest that health care workers should address aspects of mental health and energy when caring for women with physical disabilities, as these areas are often overlooked in this population. Health promotion programs aimed at these topics should be designed specifically for this population as well. PMID- 14563032 TI - Workaholism components among employees working full- and part-time. AB - This exploratory study compared Spence and Robbins' self-reported workaholism components in two samples. Data were collected in two studies from Australian psychologists and Canadian managers and professionals holding MBA degrees. As few MBA graduates and male psychologists were working part time, the results should be treated with caution. Spence and Robbins' workaholism components (work involvement, feeling driven to work, joy in work) were appropriate to part-time employees. Full-time employees tended to score higher on feeling driven to work. PMID- 14563033 TI - An evaluation of Woodard's theory of perceptually oriented hypnosis. AB - This article evaluates Woodard's theory of Perceptually Oriented Hypnosis, a welcome addition to theories of hypnosis that emphasize the centrality of subjective experiences in understanding and studying hypnosis. With a focus on subjective experience. Woodard's account of perceptually oriented hypnosis is based on elements of humanistic, client-centered, and perceptual psychology. However, we contend that improvements in the operational clarity and coverage of the theory are necessary to optimize its utility and heuristic value. We also argue that it is important for Perceptually Oriented Hypnosis to address hypnotic phenomena, e.g., individual differences in suggestibility, involuntary responding to suggestions, stability of responding over time, widely recognized as essential to understanding hypnotic experience and responding. With these refinements, we believe that Woodard's theory would have a greater potential to enrich our understanding of both the unique experiences of individuals who undergo hypnosis and of hypnosis in general. PMID- 14563034 TI - Workaholism and divorce among Australian psychologists. AB - A study of Australian psychologists compared scores on workaholism components among those who were divorced and those who were married. No differences were found. PMID- 14563036 TI - A word-association technique as a psychotherapeutic tool. AB - A way of using a word-association technique psychotherapeutically, although not experimental, has been employed as a shortcut "royal road" to understanding a presenting problem. PMID- 14563035 TI - Are students' perceptions of parental acceptance of academic dishonesty associated with its occurrence? AB - The study explored the association between the perceptions 175 Chilean business students held about their parents' acceptance of academic dishonesty and their self-reports of academic dishonesty. Regressing scores for parental acceptance onto self-reported academic dishonesty indicated it accounted for a small (2.2%) but significant amount of variance beyond demographic and academic performance variables. Effect size analysis based on structure coefficients indicated that parental acceptance was the second best predictor in the equation, suggesting that parental acceptance is a correlate that merits further study. PMID- 14563037 TI - Self-conscious emotions and criminal offending. AB - This study examined the relation of personality traits--shame-proneness, guilt proneness, and pride--on offending behavior. Using survey data from a sample of 224 college students, the construct and criterion-related validity of scales of the Shame Proneness Scale, the Test of Self-conscious Affect, and the Personality Feelings Questionnaire-2 were assessed. Regression analyses showed that self conscious emotions are important in the etiology of criminal offending. Specifically, rated pride was positively correlated with self-reported criminal activity, whereas ratings of guilt were negatively associated with offending. The relation of shame with criminality varied depending on the type of measure used to indicate proneness to shame. PMID- 14563038 TI - Longitudinal analysis of data with multiple binary category choices. AB - In many experiments, subjects mark all categories that apply when responding to a cafeteria or multiple-response question. One exact and two approximate permutation methods are described to analyze binary answers to multiple-response questions in longitudinal experimental designs, wherein the same or matched subjects respond to the same multiple-response question over two or more trials. The described methods provide probabilities, under the null hypothesis, that the multiple binary responses do not differ over trials. PMID- 14563039 TI - Bortner type A scores and basic emotions: aggression, distrustful, depression, and gregarious. AB - We examined the correlations between scores on Bortner's scale and measures of aggression, distrustful, gregarious, and depression in 1,084 employees. Prospectively over 2 yr. we investigated the relationship of Type A behavior with cardiac mortality. Subjects classified as Type A had higher mean ratings on measures of aggression and distrustful than did persons classified as Type B. Mortality was lower in patients classified as reporting Type A behavior (12%) than in the patients classified as reporting Type B behavior (22%). PMID- 14563040 TI - Students' performance in accounting: differential effect of field dependence independence as a learning style. AB - This study examined the differential moderating effects associated with field dependence-independence and perceptions of stress on students' performance after controlling for SAT Mathematics and Verbal scores as well as students' actual effort on homework. The average performance of 178 third-year accounting majors over three examinations was used to evaluate their understanding of financial accounting. The students also took the Group Embedded Figures Test. While the data indicate that the most significant variables were students' effort, SAT Verbal scores, and their perceptions of stress, these variables were differentially associated with students' performance depending upon whether the student was classified as a field-independent or field-dependent learner. PMID- 14563041 TI - Our-component model for counseling clients with traumatic childhood abuse. AB - To treat the effects of traumatic childhood abuse effectively, we propose a model which incorporates information from neurophysiological, psychoeducational, cognitive, and social work literature. The four components of the model reflect the broad range of explanations for pathology posed by researchers and also support the breadth of interventions available for use with survivors of childhood abuse. The model relies on individualized assessment and treatment related to the physiological response to abuse, faulty learning, cognitive and psychological problems, and social effects of abuse. This model contributes to the literature by providing a comprehensive framework complementary to many theoretical orientations, is useful across the helping disciplines, and appropriate in multidisciplinary settings. PMID- 14563042 TI - Documentation of demographic data. AB - Accurate identification of the demographic characteristics of subjects is crucial for clinical, research, teaching, and administrative purposes in psychology. The Demographic Coding Form is a simple, self-report tool to identify essential demographic data. Secondary categorization as "Hispanic" is obtained to facilitate comparison with the general population data. The form requests information about ethnic mixtures in subjects. Psychometric properties are being elucidated. PMID- 14563043 TI - A prescribed employee fitness program and job-related attitudes. AB - A prescribed physical fitness exercise program was initiated in a small furniture manufacturing plant. The present purpose was to investigate the effect of an intervention to change employees' attitudes and sense of well-being. The objective was to evaluate the changes in employees' attitudes (well-being) attributable to the exercise program. The method of evaluation was the survey administered to a treatment (n = 22) and a mixed (n = 30) control group. A multidimensional attitude survey questionnaire was administered to assess effects over a 6-mo. period. While some changes in job attitudes were observed for the treatment group, those were not the expected changes. Possible explanations for the findings are discussed. PMID- 14563044 TI - Reliability and factor structure of the sport anxiety questionnaire in fifth- and sixth-grade children. AB - In a sample of 178 children, the Sport Anxiety Questionnaire showed acceptable internal consistency and homogeneity. PMID- 14563045 TI - Establishing preference for unreliable reinforcement in adults with dual diagnoses. AB - We evaluated the choice responding of three adults dually diagnosed with developmental and psychiatric disabilities using concurrent schedules of reinforcement. Specifically, participants were given a choice between a response option resulting in reliable reinforcement and a response option resulting in unreliable reinforcement. Our primary purpose was to shift preference from reliable to unreliable reinforcement via the systematic presentation of stimuli during delay intervals. A second purpose was to evaluate the effectiveness of intervening stimuli in shifting preference at differing delay-to-reinforcement intervals. Preference for unreliable reinforcement was first examined in the absence of stimulus presentations during delays, at three different delay values. Next, we aimed to establish preference for unreliable reinforcement by presenting pictures of reinforcers during delays preceding unreliable reinforcement. Preference was again examined at three different delay values. In the absence of stimulus presentations during delays, participants were shown to prefer reliable reinforcement, particularly at the longer delay value. When stimuli were presented during the delays, two of the three participants preferred unreliable reinforcement, particularly the longer the delay value. These results suggest that the presentation of intervening stimuli during delays may help facilitate tolerance for unreliable reinforcement. PMID- 14563046 TI - Does psychopathology predict counseling duration? AB - Many university counseling centers have adopted case management policies in an effort to conserve limited resources. Fearing that students with more severe problems will consume too many clinical resources, many counseling centers have decided to refer such students to external agencies or providers for mental health services. However, this fear might be unwarranted because empirical research has not shown a substantial relationship between psychopathology and counseling duration. This investigation examined whether a new treatment-planning inventory, the Butcher Treatment Planning Inventory, might be useful for better understanding the relationships between various problem areas and counseling duration. Participants were new clients (students and staff members) at a university counseling center in the southeastern USA. Professional staff members and graduate students in counselor education, clinical psychology, and clinical social work provided counseling services to the participants. Lower scores on several scales predicted counseling duration. In other words, clients with lower scores on these scales (representing less psychopathology) attended more counseling sessions than clients with greater psychopathology. Therapists' background also predicted counseling duration; clients who worked with either a professional staff member or graduate student in counselor education attended more sessions than clients who worked with a graduate student in clinical psychology. The results of this study, combined with previous research, suggest that measures of psychopathology are not very useful for identifying which clients will complete long-term counseling. Until better information becomes available, actuarial tables remain the most efficient means for predicting counseling duration. PMID- 14563047 TI - Contributions to the history of psychology: CXVII. Who's who in American psychology: a citation study of introductory psychology textbooks. AB - The author indexes of 15 introductory psychology textbooks were surveyed to identify the authorities cited most frequently. The five names cited most often in descending order were Freud, Bandura, Skinner, Piaget, and James. Differences from earlier comparable citation studies are discussed. PMID- 14563048 TI - Factors influencing evaluations of web site information. AB - This study investigated the effect of first-person and third-person perceptions of web site information. Responses from a telephone survey of 226 participants in a stratified random sample indicated that (1) most participants had higher evaluations for television news than for news received on the Internet; (2) a third-person effect was present in that most respondents generally thought that other people found the Internet easier to use than they did, and that other people were more likely to believe Internet information and trust the sources of Internet information than they would. Also, (3), evaluations of information on a particular web site could be increased by providing links to other web sites on the same topic. Perhaps links to other web sites may serve as either a "referencing" function or a social confirmation function to increase evaluations of web site information. PMID- 14563049 TI - Light-hearted negative terminology used in computer education and related skill of noncomputer users. AB - Theory suggests that light-hearted but negative phrases and terminology used in computer literature and course titles may have a negative effect on noncomputer users. This study tested the effect of providing the suggestion of a negative dimension of computer ability labelled with a category of "Computer Dummy" a also the relation between computer ability and computer anxiety. The study incorporated 37 noncomputer users, 17 men and 20 women whose mean age was 52.1 yr. Scores on a self-report questionnaire indicated that 23 of the nonusers responded on the negative dimension of computer ability and included themselves in the category of "Computer Dummy." Also, belief about ability and belief about computer training were associated with computer anxiety for noncomputer users. PMID- 14563050 TI - The Kuwait University Anxiety Scale: results for 9,031 Saudi students. AB - The Kuwait University Anxiety Scale was administered to 9,031 male (n = 4,143) and female (n = 4,888) Saudis. They were students in secondary schools (n = 4.793) and university undergraduates (n = 4,238) recruited from scattered geographical regions of the Kingdom of Saudi Arabia. Ages ranged from 15 to 26 years, mean age 19.5 yr. (SD = 2.8). The scale displayed good alphas (from .85 to .88), retest reliability (from .94 to .95), as well as good criterion-related validity (from .63 to .73) against the Trait subscale of the State-Trait Anxiety Inventory. Three highly loaded factors were extracted: Cognitive/Affective, Subjective, and Somatic anxiety. Sex differences overshadow age differences as sex-related differences were significant between the age groups from 16 to 25 years, i.e., females attained higher mean scores than their male peers. Saudi students attained means similar to those of Kuwaiti students, but both groups have significantly higher mean scores than American and Spanish college students. By and large, the scale can be recommended for use in research among students in the Saudi context. PMID- 14563051 TI - Demographic, attitude, and personality differences reported by students enrolled in online versus traditional courses. AB - This study investigated demographic, personality, and attitude differences between students enrolled in online courses and students enrolled in traditional, lecture courses. Subjects were 120 education majors enrolled in required Master's of Education core courses. Students enrolled in either online (n = 51) or traditional course format (n = 69) during summer and fall semesters 2001. The two groups of students were compared on an author-designed questionnaire assessing students' demographic characteristics and attitudes toward both online and traditional instruction and the Cattell Sixteen Personality Factor Questionnaire. Analysis indicated high demographic and interpersonal similarity between students enrolled in online course formats and students enrolled in traditional, face-to face course formats. Significant differences in attitude were found between the two groups; online courses were rated higher by online students but viewed with considerable ambivalence by students enrolled in traditional section courses. PMID- 14563052 TI - Significance of violence in stories of young children. AB - Stories told by 72 5-yr.-old children were scored for themes of violence and correlated with their mothers' reports of aggressive or fearful behaviors. Values for the entire sample were not significant. However, when data for Hispanic children were removed from the sample the correlation of violent stories to ratings of children's aggression was significant (r = .27) and for a count of violent words r = .30. The correlation of violent words to fearfulness was .28 and for girls only .35. The count of violent words was also correlated with African-American children's ratings of fearfulness (r = .64). PMID- 14563053 TI - Membership in voluntary associations and rates of suicide in the provinces of Canada. AB - Applying Lester's method to data from 10 Canadian provinces, the 1997 and 1999 age-standardized rates of suicide were not associated with the extent (%) to which Canadians participated in voluntary associations. PMID- 14563054 TI - Fusion or confusion in obsessive compulsive disorder. AB - Inferential confusion occurs when a person mistakes an imagined possibility for a real probability and might account for some types of thought-action and other fusions reported in obsessive-compulsive disorder. Inferential confusion could account for the ego-dystonic nature of obsessions and their recurrent nature, since the person acts "as if" an imagined aversive inference is probable and tries unsuccessfully to modify this imaginary probability in reality. The clinical implications of the inferential confusion model focus primarily on the role of the imagination in obsessive-compulsive disorder rather than on cognitive beliefs. PMID- 14563055 TI - Loneliness and health-related variables in early adolescents: an extension. AB - The purpose of this study was to extend knowledge about loneliness and health related variables. In classrooms, 135 early adolescents responded to measures of loneliness, symptom patterns, and perceived health status. Statistically significant correlations were found for loneliness scores with ratings for symptom patterns (.44) and with ratings for perceived health status (-.46). PMID- 14563056 TI - Characteristics of readmitted psychiatric inpatients. AB - This study examined sociodemographic, diagnostic, psychological, and episode based variables in a sample of 130 psychiatric patients admitted to treatment at least twice in a 6-yr. period. Short length of initial hospitalization (r = -.30, p < .01) and younger age on initial admission (r = -.20, p < .05) were significantly correlated with frequent hospital admissions. Scores on four of the subscales of the WAIS-R were significantly correlated with readmission, confirming that patients who have fewer cognitive resources are at risk of frequent admissions. A multiple regression analysis combining variables to predict readmission accounted for only 12% of the common variance (r128 = .34, p < .01), however, indicating that a prediction equation with these variables has limited clinical utility. PMID- 14563057 TI - Flirting and energetic arousal. AB - 76 college students were administered a self-reported energy scale and a flirtation inventory. Analysis indicated a significant relationship such that low scores on energy level were associated with low flirtation related activities, as well as low self-ratings on attractiveness and other variables. PMID- 14563058 TI - Pilot study comparing market orientation culture of businesses and schools of business. AB - A market orientation culture has been described as one that blends an organization's commitment to customer value with a process of continuously creating superior value for customers. Developing such a culture is further described as (1) obtaining information about customers, competitors, and markets, (2) examining the gathered information from a total organizational perspective, (3) deciding how to deliver superior customer value, and (4) implementing actions to provide value to customers. A market orientation culture focuses on the customer, identifies issues in the competitive environment, and coordinates all functional areas to achieve organizational objectives. Research has found businesses with higher market orientation are more successful in achieving organizational objectives. The measurement of market orientation within businesses has been empirically tested and validated. However, empirical research on market orientation in nonprofit organizations such as universities has not been examined. This study investigated market orientation within the university setting, specifically Schools of Business Administration, and compared these data with previously published data within the business sector. Data for comparative purposes were collected via a national survey. Hypothesis testing was conducted. Results indicated significantly lower market orientation culture within the schools of business as reported by AACSB Business School Deans vis-a-vis managers of business enterprises. PMID- 14563059 TI - Relations of perceived ethnic disparity to involvement in ethnic and host cultures. AB - Among 124 single foreign students residing in the USA, this study examined the relations between perceived ethnic disparity, consisting of physical, cultural. and linguistic dimensions, and ethnic and host culture involvement. The Ethnic Disparity Perception Inventory, a 44-item Likert scale devised to measure perceived ethnic disparity, showed perceived physical disparity (r = .60) and cultural disparity (r = .51) were positively related to ethnic involvement. Perceived cultural disparity (r = -.22) and linguistic disparity (r = -.27) were negatively associated with involvement in the host culture. Multiple regression analysis indicated that, although the findings held for non-White respondents, for White respondents (n = 39) perceived physical disparity (beta = .54), not cultural disparity, was a significant predictor of ethnic involvement (Adjusted R2 = .268) and perceived cultural disparity (beta = -.37), not linguistic disparity, was a significant (negative) indicator of host culture involvement (Adjusted R2 = .110). PMID- 14563060 TI - Sex differences in recall of real or nonsense words. AB - Women perform better than men on tests of verbal memory, but the nature of this advantage has not been precisely established. To examine whether phonemic memory is a factor in the female advantage, we presented, along with other verbal memory tasks, one containing nonsense words. Overall, there was the expected female advantage. However, an examination of the individual tests showed female superiority in recall of the real words but not the nonsense words. PMID- 14563061 TI - Application of Herek's attitudes toward lesbians and gay men scale in The Netherlands. AB - A national sample of 921 respondents from the Dutch population completed the translated, slightly modified version of Herek's Attitudes Toward Lesbians and Gay Men Scale, originally developed in the USA for the assessment of attitudes toward lesbians and gay men. The present study was the first attempt to test the applicability of the scale in the Netherlands. In line with previous American findings, all scale items loaded highly on one factor. The scores also correlated with another measure of attitudes toward lesbians and gay men, namely, 'denial of equal rights' (r = .76), and with 'traditional view of women' (r = .48), 'importance of traditional marriage' (r = .53), and 'authoritarianism' (r = .31). Data support the scale's acceptable reliability and construct and discriminant validity. PMID- 14563062 TI - Obtaining funding for new researchers in psychology. AB - A mentor is the key person to assist a student who wishes to become an independent investigator. The federal government provides long-term funding for decades of research by an investigator, and short-term funding for years of transition from student to investigator is available from both governmental and private agencies. Grants designed for men and women and various ethnic groups are valuable resources. Courteous acknowledgment of sponsorship by the grantee facilitates continued grants from the funding source. PMID- 14563063 TI - A cross-national perspective on school bullying in Northern Ireland: a supplement to Smith, et al. (1999). AB - There is great value in exploring the prevalence of school bullying from a cross national perspective. Smith, Morita, Junger-Tas, Olweus, Catalano, and Slee in 1999 presented a cross-national perspective on the nature, prevalence, and correlates of school bullying that encompassed a wide range of countries. However, Northern Ireland was not included, despite potentially being an important country to include, given its volatile social, ethnic, and religious history--leading to the concern that the population has become somewhat habituated to low level aggression. Thus, the present paper provides a review of the current literature on school bullying in the Northern Ireland school system. Evidence presented suggests that the incidence of school bullying in Northern Ireland may be higher than that in the rest of Ireland and the United Kingdom. PMID- 14563065 TI - Chinese version of the separation-individuation inventory. AB - The importance of the separation-individuation process in object relations theory is well known in disciplines of psychology, counseling, and human development. Based on the Separation-Individuation Inventory of Christenson and Wilson, which measures the manifestations of disturbances in this process, a Chinese version of the inventory was developed. For college students Cronbach coefficient alpha was .89, and test-retest reliability over 28 days was .77. The scores of the inventory had positive correlations with both the number of borderline personality characteristics and the Individualism-Collectivism Scale, respectively. Also, the mean score on the inventory of patients diagnosed with borderline personality disorder was significantly higher than that of the two normal control groups (ns = 564). Thus the inventory possessed satisfactory construct validity. Cultural differences regarding the separation-individuation process need to be investigated further. PMID- 14563064 TI - Perceived locus of control as a function of parental physical punishment among a group of Iranian children. AB - A comparison of scores on locus of control by two groups of children, 89 who had experienced parental physical punishment and 174 who had not experienced parental physical punishment, showed that children with experience of physical punishment scored significantly more external than children who had not experienced physical punishment. The results are discussed in relation to previous studies, and some educational implications noted. PMID- 14563066 TI - Family contexts, individualism-collectivism, and adolescents' aspirations for their futures. AB - Relations were examined among family contexts, parents' individualistic collectivistic value orientations, and adolescents' aspirations. Data were collected from 456 Australian families using two surveys when the subjects were 11 and 16 years old, respectively. Findings indicated that family ethnicity had large associations with parents' individualistic-collectivistic value orientations, but that these orientations were not associated with adolescents' aspirations, after taking into account differences in family contexts. PMID- 14563067 TI - Diminishing incidence of Internet child pornographic images. AB - Indecent images of children posted to web sites and newsgroups over a 4-yr. period were sampled. A significant decline in the number of such images posted was observed, probably accounted for by the pressure of groups opposed to the distribution of such exploitive material. PMID- 14563068 TI - Empirical research and logotherapy. AB - The purpose of this paper was to outline suggestions for future logotherapy applications and research. Empirical studies are available on logotherapy constructs, but additional research is sorely needed. The psychometrics of logotherapy measures need to be expanded. Carefully conducted experimental studies may aid in refining long-standing logotherapy concepts and supporting or refuting new ones. PMID- 14563069 TI - Identifying careless responses in questionnaire research: a serendipitous finding. AB - The present study describes a possible method by which potentially meaningless responses to questionnaires can be easily identified. Given an inadvertent mistake in the design of a questionnaire packet, we found that 10% of respondents provided invalid responses to items. PMID- 14563070 TI - Proceedings of the Conference on Social Determinants of Health and Disease. November 2002, Chicago, Illinois, USA. PMID- 14563071 TI - Understanding social inequalities in health. AB - A prominent feature of health in all industrialized countries is the social gradient in health and disease. Many observers believe that this gradient is simply a matter of poor health for the disadvantaged and good health for everyone else, but this is an inadequate analysis. The Whitehall Study documented a social gradient in mortality rates, even among people who are not poor, and this pattern has been confirmed by data from the United States and elsewhere. The social gradient in health is influenced by such factors as social position; relative versus absolute deprivation; and control and social participation. To understand causality and generate policies to improve health, we must consider the relationship between social environment and health and especially the importance of early life experiences. PMID- 14563072 TI - Secular trends in physiological capital: implications for equity in health care. AB - Over the past three centuries, there has been a rapid accumulation of physiological capital in Organisation for Economic Cooperation and Development (OECD) countries. Enhanced physiological capital is tied to long-term reduction in environmental hazards and to the conquest of chronic malnutrition, since both nutritional status and the quality of the external and intrauterine environments appear to be linked to the quality of organ development and to the onset of chronic diseases later in life. Data on heights and birth weights suggest that physiological capital has become more equally distributed, thereby reducing socioeconomic disparities in the burden of disease. These developments have a number of health care policy implications: (1) enhanced physiological capital has done more to reduce inequities in health status than has wider access to health care; (2) the main contribution of more advanced medical treatment so far has been to retard depreciation in individuals' physiological capital; (3) prenatal and early childhood care and environmental issues are key for interventions aimed at enhancing physiological capital and at affecting its rate of depreciation; (4) lifestyle change is the most important issue affecting health equity in rich countries; and (5) greater access to clinical care should be promoted through aggressive outreach, since expanded insurance coverage by itself is inadequate. PMID- 14563074 TI - The neighborhood context of well-being. AB - Health-related problems are strongly associated with the social characteristics of communities and neighborhoods. We need to treat community contexts as important units of analysis in their own right, which in turn calls for new measurement strategies as well as theoretical frameworks that do not simply treat the neighborhood as a "trait" of the individual. Recent findings from the Project on Human Development in Chicago Neighborhoods support this thesis. Two major themes merit special attention: (1) the importance of collective efficacy for understanding health disparities in the modern city; and (2) the salience of spatial dynamics that go beyond the confines of local neighborhoods. Further efforts to explain the causes of variation in collective processes associated with healthy communities may provide innovative opportunities for preventive intervention. PMID- 14563073 TI - Social isolation and health, with an emphasis on underlying mechanisms. AB - Social isolation is a potent but little understood risk factor for morbidity and mortality, and its negative consequences are most profound among the elderly, the poor, and minorities, some of the fastest growing segments of the U.S. population. A steadily increasing number of people are living alone and are therefore more likely to experience social isolation. We discuss four mechanisms- attractiveness, health behavior, stress, and repair and maintenance--by which perceived social isolation might affect health. Our studies show that neither attractiveness nor health behaviors differ as a function of social isolation, but stress and repair and maintenance do seem to be factors. While socially isolated young adults did not report more frequent everyday stressors, they rated everyday events as more intensely stressful. They were also more likely to report passively coping with stressors, and to show greater vascular resistance, a mechanism of blood pressure control previously associated with passive coping and a risk factor for hypertension. Finally, they exhibited less efficacious repair and maintenance of physiological functioning, including slower wound healing and poorer sleep efficiency. These mechanisms have implications for designing appropriate interventions. We advocate a national health care plan that promotes preventive medicine, recognizes the significance of stress-related disorders, and supports the maintenance of social connections across the life span. PMID- 14563075 TI - Are there social determinants of health and disease? AB - The concept of a determinant is tied to the idea of a mechanism for action. Ideas from epidemiology, particularly the epidemiologic triad of agent, host, and environment, can help to make sense of factors that affect the absence of disease or that interfere with a mechanism that alters health. However, assembling convincing evidence for the existence of social determinants of health is a challenge, in part because of the difficulties of bridging the social and biological realms. While social contexts are measured using aggregates of individuals, disease and dysfunction occur at the individual level, leading to difficult problems of ecological inference. Although social factors have been shown to be associated with differences in mortality from specific causes, these factors account for only a small portion of the mortality from any individual cause. This suggests that the pathways through which social factors influence health are affected by their interactions with other factors. PMID- 14563076 TI - Synthesis of scientific disciplines in pursuit of health: the Interactive Biopsychosocial Model. AB - Twenty-five years ago, George Engel proposed a challenge to the biomedical model and its limited view of disease as biologically rooted. Building on Engel's work, we present the Interactive Biopsychosocial Model (IBM). The IBM argues for a reorientation in biomedicine where research, education, and clinical practice: (1) address health in addition to illness; (2) aim to decipher interrelated biophysical, psychocognitive, and social processes in health and disease, rather than seek a single root cause; and (3) take into account the social networks of the individual to achieve, maintain, and maximize health and well-being for individuals, their significant others, and society. Based on an interdisciplinary collaboration of medical and social scientists, this paper demonstrates the application of the IBM to understanding and generating hypotheses about the longitudinal relationship between sexuality and health, and sexuality and chronic illness (diabetes mellitus) at older ages. The model provides a dynamic, dyadic, framework for building scientific hypotheses about the etiologies and consequences of health, well-being, and disease throughout the life course. PMID- 14563077 TI - The long-run growth in obesity as a function of technological change. AB - This paper analyzes the forces contributing to the worldwide long-run rise in obesity and the role of public interventions in affecting its continued growth. A growth in obesity in a population must result from the growth of calorie consumption outpacing the growth of physical activity. Yet historically in developed countries, obesity has grown with modest rises in calorie consumption and with a substantial increase in both exercise and dieting. We consider the economic incentives that give rise to the long-run growth in obesity by stimulating intake of calories at the same time as discouraging the expending of calories on physical activity, whether in work or leisure. We argue that technological change provides a natural interpretation of the long-run growth in obesity, that it predicts that the effect of income on obesity changes from positive to negative with economic development, and that it implies that technological change may not continue to raise weight. We discuss the positive and normative impacts of direct and indirect public interventions to reduce obesity. PMID- 14563078 TI - Diminishing returns? The costs and benefits of improving health. AB - In this paper, we develop and apply an economic framework for valuing improvements in health, based on Murphy and Topel (2002). Past improvements in health and longevity have had enormous social value. By our estimation, the contributions to the economic well-being of longer lifetimes have been worth about dollar 73 trillion since 1970, or about dollar 2.6 trillion per year. The average 50-year-old man gained additional life-years worth roughly dollar 350,000, while a 50-year-old woman gained about dollar 180,000. Looking ahead, prospective benefits from further progress are also large: a 10 percent reduction in mortality from heart disease would be worth over dollar 4 trillion, and even a 1 percent reduction in cancer deaths would be worth over dollar 400 billion. These values suggest that public investments in basic medical research may yield huge social returns, but distortions in the allocation of medical care and in research incentives may yield future benefits that are smaller than the costs of achieving them. While the average net health benefits of increased medical expenditures are large, costs may have greatly exceeded benefits in certain periods and age groups. PMID- 14563079 TI - Is medical research cost effective? Response to Murphy and Topel. AB - The paper explores the relationship between medical innovation and cost of treatment. The methodology used by Murphy and Topel to calculate the statistical value of a life is examined. The authors discuss possible confounding factors, such as the general trend of decreasing mortality, and consider the possibility that non-medical sources, especially the decrease in tobacco use, may explain much of the decrease in mortality rates. The authors suggest an alternative to Murphy and Topel's population-level model for assessing the benefits of medical technology, based on Lewis Thomas' categorization of three levels of technology: non-technology ("caring"), half-way technology (does not reverse or prevent the underlying problem), and high technology (preventative or curative). The categories are applied to the example of type 1 diabetes mellitus, in which quality of life and mortality improvements are clearly the result of technological changes and not behavioral modifications. The authors emphasize the cost-effectiveness of high technology, which is based on medical research advances. PMID- 14563080 TI - Let the shoemaker stick to his last: a defense of the "old" public health. AB - The traditional forms of public health law were directed largely toward communicable diseases and other externalities, such as pollution, with negative health impacts. The more modern view treats any health issue that affects large numbers of individuals as one of public health, which would include such problems as obesity and diabetes. This paper examines the constitutional evolution of the public health principle from the narrower to the broader conception. It then argues that the narrower conception better defines the appropriate scope of coercive government intervention than does the broader definition, which could easily authorize interventions in economic affairs whose indirect effects are likely to reduce overall social wealth and freedom, and with it the overall health of the population. PMID- 14563081 TI - The politics of public health: a response to Epstein. AB - Conservatives are taking aim at the field of public health, targeting its efforts to understand and control environmental and social causes of disease. Richard Epstein and others contend that these efforts in fact undermine people's health and well-being by eroding people's incentives to create economic value. Public health, they argue, should stick to its traditional task--the struggle against infectious diseases. Because markets are not up to the task of controlling the transmission of infectious disease, Epstein says, coercive government action is required. But market incentives, not state action, he asserts, represent our best hope for controlling the chronic illnesses that are the main causes of death in industrialized nations. In this article, we assess Epstein's case. We consider his claims about the market's capabilities and limits, the roles of personal choice and social influences in spreading disease, andthe relationship between health and economic inequality. We argue that Epstein's critique of public health over-reaches, oversimplifies, and veils his political and moral preferences behind seemingly objective claims about the economics of disease control and the determinants of disease spread. Public health policy requires political and moral choices, but these choices should be transparent. PMID- 14563082 TI - Private wealth and public health: a critique of Richard Epstein's defense of the "old" public health. AB - This article is a response to Richard Epstein's recent legal and economic critiques of contemporary public health law. The power of Epstein's critical position stems from the convergence of three problems, each with distinctive intellectual histories: (1) an age-old debate within political liberalism about the proper relationship of individual right and state power; (2) a somewhat more recent biomedical policy contest over the necessary scale and scope of public health interventions in modern societies; and (3) an ongoing legal jurisprudential argument about the overall trajectory and implications of American constitutional history. My critique of Epstein's position follows this same tripartite format, moving from an initial discussion of the history of liberalism to the history of public health and, finally, to a critique of Epstein's general legal history of state regulation in America. PMID- 14563083 TI - The scope and limits of public health law. AB - Public health law needs to differentiate more clearly between public health analysis and public health authority, that is, between the scientific boundaries of epidemiology and tile legal and regulatory boundaries of public health. These boundaries matter because public health law confers tremendous authority on government officials, including some powers that are justified only in situations of extreme urgency. The extent of this legal power explains the inclination of activists to apply public health paradigms to various social problems beyond the traditional origins of public health law; once a causal connection to a widespread health problem is identified, it falls within the authority of public health officials to take whatever actions are necessary to eradicate the threat to health. This approach might lead to dangerous conditions in which public health officials overstep the proper bounds of public health law, even though they arguably are continuing to exercise proper analytical tools for understanding public health problems. Public health authority should remain grounded in traditional conceptions of disease, which depend on a specific agent or behavior that threatens health in a direct and clear manner, and for which a targeted and effective remedy that requires collective action is available. It should not extend to diseases viewed as resulting from social, economic, and political conditions. PMID- 14563085 TI - Obstetric analgesia: back to the future? PMID- 14563084 TI - Technological advancement and human reproduction: where can we go or where do we want to go? PMID- 14563086 TI - Analysis of uterine contractions: a dynamical approach. AB - The development of suitable techniques for quantifying mechanical and electrophysiological aspects of uterine contractions has been an active area of research. The uterus is a physiological system consisting of a large number of interacting muscle cells. The activity of these cells evolves with time, a trait characteristic of a dynamical system. While such complex physiological systems are non-linear by their very nature, whether this non-linearity is exhibited in the external recording is far from trivial. Traditional techniques such as spectral analysis have been used in the past, but these techniques implicitly assume that the process generating the contractions is linear and hence may be biased. In this tutorial review, a systematic approach using a hierarchy of surrogate algorithms is used to determine the nature of the process generating the contractions produced during labor. The results reveal that uterine contractions are probably generated by non-linear processes. The contraction segments were obtained through simultaneous recordings of the electrical and magnetic signals corresponding to the electrophysiological activity of the uterus and then analyzed. The electrical activity was recorded by placement of non invasive electrodes onto the maternal abdomen and magnetic activity was recorded non-invasively using a superconducting quantum interference device (SQUID). PMID- 14563087 TI - Delivery at 34 weeks is more costly than at 35 weeks in pregnancies with premature rupture of membranes. AB - OBJECTIVE: To assess the relative risks and costs of delivery at 34 vs. 35 weeks with preterm premature rupture of membranes (PROM). STUDY DESIGN: Retrospective study of singleton gestations with preterm PROM over a 5-year period. Patients who delivered at 34 vs. 35 weeks were compared. RESULTS: Sixty-five patients were identified in each group. There were no significant differences in maternal demographics or complications. Direct costs of mother-infant pairs at 34 weeks were significantly higher (dollars 6687 +/- 4273 vs. dollars 4089 +/- 3037, p = 0.0001). Newborn intensive care unit stay was significantly longer for infants born at 34 weeks (8.4 +/- 7.1 vs. 3.5 +/- 4.6 days, p < 0.0001). CONCLUSIONS: There was a significant increase in neonatal interventions, neonatal length of stay and cost for infants delivered at 34 vs. 35 weeks. Prospective data are necessary to evaluate the actual maternal and neonatal morbidity incurred by delay of delivery. PMID- 14563088 TI - Middle cerebral artery blood flow in the fetus and central nervous system complications in the neonate. AB - OBJECTIVE: To evaluate the ability of blood flow (S/D) ratios in the fetal middle cerebral artery (MCA) to predict the occurrence of central nervous system (CNS) complications (bleeding and/or ischemia) in the neonate, and compare MCA flow to other tests of fetal well-being. STUDY DESIGN: This was a retrospective, observational, cross-sectional clinical outcome study of high-risk patients, evaluated in the Perinatal Center of the author's institution, who underwent antenatal testing with non-stress test (NST) and Doppler studies (including studies of the fetal umbilical artery and MCA) within 1 week of delivery. MCA flow was converted to multiples of the mean, and appropriate parametric and non parametric statistics were used to compare MCA flow to the presence or absence of CNS complications. Comparisons were also made for the other tests of fetal well being. RESULTS: CNS complications occurred only in infants who were delivered at less than 37 weeks of gestation (n = 14/131) and the analysis was limited to these 131 patients. Univariant analysis showed that only the NST correlated with CNS complications (OR = 5.46 (1.48-22.16)) and logistic regression confirmed the association. Neither increased diastolic flow in the MCA nor the presence of shunting correlated with CNS complications. CONCLUSIONS: This study confirms previous reports that fetal shunting, with decreased resistance and increased flow in the CNS, is a benign adaptive mechanism for the fetus under stress and not an increased risk for CNS complications. A non-reactive NST, however, is a very late sign of fetal compromise and is significantly associated with the risk of developing CNS complications. PMID- 14563089 TI - Induction of labor with vaginal prostaglandin E2. AB - OBJECTIVE: To determine the effects of labor induction with vaginal prostaglandin E2 (PGE2) on mode of delivery in post-term pregnancies and to identify possible predictors of successful vaginal delivery. METHODS: The study group consisted of 211 women with post-term pregnancy (gestational age more than a full 41 weeks) admitted for induction of labor with vaginal applications of PGE2. Findings were compared with women with spontaneous onset of labor (group 2, n = 510). Women with previous Cesarean section were excluded from both study and control groups. RESULTS: The rate of Cesarean section was significantly higher in the study group (19.4%) than in the control group (9%). A logistic regression model was used to control for maternal and gestational age, gravidity, parity, nulliparity rate, number of used PGE2 tablets, oligohydramnios and delivery weight as possible predictors of Cesarean section (R2 = 0.56, p < 0.001). On stepwise analysis, nulliparity (OR 45.2, 95% CI 2.93-695.5), number of PGE2 applications (OR 2.79, 95% CI 1.04-7.52) and maternal age (OR 1.23, 95% CI 1.01-1.50) were independently and significantly associated with increased risk of Cesarean section. PGE2 induction was a significant independent predictor of Cesarean section (OR 2.3, 95% CI 1.3-3.9). CONCLUSION: PGE2 induction of labor was successful in approximately 80% of patients with post-term pregnancy, with apparently no serious maternal or fetal complications. Risk factors intrinsic to the patient, and labor induction itself, were the reasons for excess Cesarean deliveries in women with prolonged pregnancies. PMID- 14563090 TI - Severity of obstetric complications and risk of adult schizophrenia in male patients: a case-control study. AB - OBJECTIVE: Obstetric complications may be an etiologically important factor in the development of schizophrenia. The aim of this study was to evaluate whether the risk for developing schizophrenia in adult life is increased in individuals with more severe obstetric complications at birth. METHODS: To this end, mothers were interviewed to gather data about obstetric complications. The 'midwife protocol' of Parnas and colleagues was used to quantify the presence and entity of obstetric complications. We studied the frequency distribution and the severity of obstetric complications in 64 male DSM IV schizophrenic patients. The genetic load was reduced by using 81 brothers who were not psychiatric patients as controls. Odds ratios for the effects of obstetric complications, maternal age, birth order and birth weight were calculated using conditional logistic regression. RESULTS: The only factor found to have a significant effect on the risk of schizophrenia was the overall measure of obstetric complications at birth. The history of obstetric complications was higher in schizophrenic patients than in their siblings. CONCLUSION: The results seem to confirm the hypothesis that obstetric complications may contribute to increased vulnerability to the disease, in addition to genetic risk factors. PMID- 14563091 TI - Stillbirth in obstetric practice: report of survey findings. AB - OBJECTIVE: Stillbirth affects a large portion of the population and results in mortality rates comparable to those of preterm delivery and sudden infant death syndrome combined. Despite the large burden, little information is available to offer patients regarding etiology, treatment or prevention for a subsequent pregnancy. METHODS: We surveyed a sample of Fellows of the American College of Obstetricians and Gynecologists to determine the practice patterns in the management of stillbirth. RESULTS: The majority of Fellows agreed on the definition of stillbirth; however, their approach to treatment and prevention varied. A majority of Fellows believed that research on understanding stillbirth was of national importance. CONCLUSIONS: A comprehensive educational effort to include current knowledge regarding causes and management, standardized diagnostic procedures, death registration and case review is recommended to improve obstetric care of those with a stillbirth. PMID- 14563092 TI - Maternal smoking during pregnancy improves the anatomy of the hip joint in the female neonate. AB - OBJECTIVE: Because the greater frequency of developmental dysplasia of the hip in the female could have an endocrine etiology, and because maternal smoking in pregnancy causes fetal endocrine disequilibrium, we investigated the anatomy of the hip in neonates of smoking and non-smoking mothers. METHODS: Clinical and sonographic examination was performed on 2066 hips of 521 male and 512 female neonates. In 48 male and 53 female neonates, the mothers smoked during pregnancy. RESULTS: The mean +/- SD alpha angle in the male neonates of the non-smoking mothers was 62.3 degrees +/- 5.1 degrees and of the smoking mothers 62.1 degrees +/- 4.9 degrees (p = 0.7). In the female neonates of the non-smoking and the smoking mothers, it was 60.7 degrees +/- 5.3 degrees and 61.9 degrees +/- 4.8 degrees, respectively (p = 0.02). The difference between the male and the female neonates of the non-smoking mothers was significant (p < 0.000001), but there was no significant difference between the female neonates of the smoking mothers and the male neonates (p = 0.5). Among the female neonates whose mothers were non smokers, the number of those with hip type IIa or worse was significantly greater than among the female neonates whose mothers were smokers. The clinical findings were in agreement with the sonographic findings. CONCLUSIONS: Maternal smoking during pregnancy reduces the frequency of sonographic and possibly clinically detected hip dysplasia in female but not in male neonates. Nonetheless, because smoking causes numerous adverse effects on the fetus and child, it is contraindicated during pregnancy. PMID- 14563093 TI - Amniotic fluid levels of immunoreactive monocyte chemotactic protein-1 increase during term parturition. AB - OBJECTIVE: Parturition is characterized by an influx of inflammatory cells into gestational tissues, a phenomenon conducive to increased myometrial contractility, cervical ripening and decidual/membrane activation. Monocyte chemotactic protein-1 (MCP-1), a potent chemoattractant and activator of monocytes/macrophages, is expressed in gestational tissues and, thus, may participate in the final common pathway of labor. This study was undertaken to determine whether the amniotic fluid concentrations of immunoreactive MCP-1 are altered with gestational age or spontaneous labor at term with and without prelabor rupture of the gestational membranes. We also sought to identify intrapartum differences in the concentrations of immunoreactive MCP-1 between the upper and lower amniotic fluid compartments. METHODS: A cross-sectional study was conducted to assess the concentrations of immunoreactive MCP-1 in amniotic fluid. Amniotic fluid samples were obtained from 225 women as follows: (1) women undergoing mid-trimester (14-18 weeks of gestation) amniocentesis for genetic indications, whose pregnancy outcome was normal (n = 84); (2) women in labor (n = 52) and not in labor (n = 31) at term, with intact gestational membranes; (3) women with rupture of the gestational membranes in labor (n = 18) and not in labor (n = 26), at term; and (4) women in labor at term for whom paired amniotic fluid samples were obtained through transvaginal and transabdominal amniocenteses (n = 14). Immunoreactive MCP-1 was assessed with a specific and sensitive immunoassay that had been validated for amniotic fluid. Non-parametric statistics were used for analysis. RESULTS: Immunoreactive MCP-1 was detected in all amniotic fluid samples. Spontaneous human parturition was associated with a significant increase in the amniotic fluid concentrations of immunoreactive MCP-1 (not in labor: median 595 pg/ml, range 183-3579 pg/ml vs. in labor: median 862 pg/ml, range 183-9609 pg/ml; p = 0.01). The median amniotic fluid concentrations of immunoreactive MCP-1 were significantly higher in the lower amniotic fluid compartment than in the upper amniotic fluid compartment (lower compartment: median 2913 pg/ml, range 1360-17080 pg/ml vs. upper compartment: median 1603 pg/ml, range 1070-8062 pg/ml; p = 0.004.). Spontaneous rupture of the gestational membranes at term was not associated with a significant change in the amniotic fluid concentrations of immunoreactive MCP-1. CONCLUSIONS: Immunoreactive MCP-1 is a physiological constituent of the amniotic fluid. The amniotic fluid levels of immunoreactive MCP-1 increase during spontaneous labor at term. A topographic difference in the concentration of immunoreactive MCP-1 was observed in the amniotic cavity, with higher concentrations being noted in the lower amniotic fluid compartment, as compared with the upper amniotic fluid compartment. These findings support the hypothesis that MCP-1 may play a role in the final common pathway of spontaneous labor. PMID- 14563094 TI - The effect of antibiotic therapy on intrauterine infection-induced preterm parturition in rabbits. AB - OBJECTIVE: The purpose of this study was to determine whether early antibiotic administration to pregnant rabbits with intrauterine infection could prevent preterm delivery and perinatal mortality. STUDY DESIGN: Under hysteroscopic guidance, pregnant rabbits at 70% gestation (21 days) were allocated to three groups: (1) control group, transcervical inoculation of 0.2 ml phosphate-buffered saline (n = 16); (2) infection group, transcervical inoculation of 0.2 ml of 10(5) colony-forming units (CFU) of Escherichia coli (n = 21); (3) infection and antibiotics group, transcervical inoculations of 0.2 ml of 10(5) CFU of E. coli and ampicillin-sulbactam 150 mg/kg every 8 h intramuscularly (n = 32). To examine the consequences of treatment delay, animals in the latter group were subdivided to receive antibiotics at different time intervals of 0, 6, 11 and 18 h after bacterial inoculation. The intervals from bacterial inoculation to delivery and litter survival were documented. Systemic (rectal) temperatures were recorded at 4 h intervals through the first 36 h and every 12 h until delivery. A p value of < 0.05 was considered significant. RESULTS: All rabbits inoculated with E. coli without antibiotic treatment delivered prematurely. The median inoculation-to delivery interval was significantly shorter in the infected group than in the control group (median 32 h, range 14.9-76.5 h vs. median 219 h, range 173-246 h, respectively; p < 0.0001). Antibiotic administration within 12 h of inoculation, but not after 18 h, increased duration of pregnancy (by reducing the rate of preterm delivery) and neonatal survival (0% vs. 71%; p < 0.0001). The mean temperatures at delivery of animals whose treatments began at 6 and 11 h post inoculation were significantly lower than those untreated with antibiotics or those treated at 18 h post-inoculation (p < 0.0001 for each comparison). CONCLUSIONS: Antibiotic administration can prolong pregnancy and reduce perinatal mortality if administered early (within 12 h of microbial inoculation) in a rabbit model of ascending intrauterine infection. PMID- 14563095 TI - Antenatal detection of mosaic trisomy 9 by ultrasound: a case report and literature review. AB - This paper presents a fetus with mosaic trisomy 9 diagnosed by chorionic villus sampling and confirmed by cordocentesis, and compares this case with published cases in order better to define the ultrasound markers confined to trisomy 9 syndrome. Detailed fetal ultrasound examination was carried out, revealing shortened femur, placental cysts and oligohydramnios. All published trisomy 9 cases with abnormal ultrasound findings were extracted from the MEDLINE database in the period from 1973 to 2002. We found 12 non-mosaic and 13 mosaic cases, including our case. The most frequent ultrasound abnormalities included characteristic cardiac, skeletal, craniofacial and central nervous system malformations. Intrauterine growth restriction and single umbilical artery were prevalent non-specific findings in both non-mosaic and mosaic groups. Parental chromosomal variations, as in our case, were not uncommon findings. When a fetus shows structural anomalies suggesting the presence of trisomy 9, karyotyping should be performed on both chorionic villi or amniocytes and fetal blood lymphocytes to enable a correct diagnosis to be made. PMID- 14563096 TI - A biopsychosocial approach to premenstrual dysphoric disorder. AB - Though epidemiological data is difficult to collect, existing evidence indicates that there is a small but significant population of women in whom premenstrual symptoms, and particularly affective symptoms, severely impair functioning. Although PMDD is predominantly regarded as a biologically based illness, there is strong evidence that variables such as life stress, history of sexual abuse, and cultural socialization are important determinants of premenstrual symptoms. In diagnosing and treating PMDD patients, attention to biological and sociocultural variables is recommended. PMID- 14563097 TI - Assessment and treatment of depression during pregnancy: an update. AB - Depression occurs commonly during pregnancy, and women with recurrent depression are at particularly high risk for depressive illness in this setting. Though the use of psychotropic medications during pregnancy raises concerns, there are data to support the use of certain antidepressants, including fluoxetine and the tricyclic antidepressants. Data on the newer SSRI antidepressants is gradually accumulating and is encouraging. None of the SSRIS or TCAs have been associated with an increased risk of congenital malformation. However, information on the long-term neurobehavioral effects of these medications still remains limited. As depression during pregnancy carries risk for both the mother and child, it is crucial to diagnose depression in this setting and to provide appropriate treatment strategies. Further data on nonpharmacologic and pharmacologic strategies is needed to aid in the treatment of this challenging clinical population. The clinician must weigh the relative risks of various treatment options and take into account individual patient wishes. Such a process will lead to thoughtful treatment choices, which with close clinical follow-up can minimize the risk for maternal morbidity. PMID- 14563098 TI - Assessment and treatment of hot flushes and menopausal mood disturbance. AB - More than 1 million women are expected to reach menopause each year, many of whom will experience hot flushes and other neuropsychological symptoms that may diminish their quality of life. Hot flushes are the core symptoms that reflect the brain's response to the changing hormonal milieu of the menopause transition, particularly to the rapidly fluctuating and falling levels of estradiol. The physical symptoms of hot flushes and the associated changes in sleep, mood, and cognition will lead many women to seek medical care. It is critical to understand the interrelationship of hot flushes and other neuropsychological symptoms of the menopause transition so that treatment priorities can be established. For example, if sleep disruption explains most daytime neuropsychological problems in women with hot flushes, treating insomnia should be considered a priority. Alternatively, mood, cognition, and quality of life may be disturbed independent of sleep problems. In such a situation, each symptom should be evaluated separately from any assessment of sleep. As recent data from the WHI establish the risks of long-term HRT use, concern about using HRT, even as a short-term intervention, has increased substantially. Although HRT remains the first-line treatment for hot flushes, the WHI findings have drawn attention to nonhormonal treatments of hot flushes and other menopausal symptoms. Growing evidence to support the efficacy of serotonergic antidepressants and other psychoactive medications in the treatment for hot flushes suggests that nonhormonal interventions will prove important alternatives to HRT. As further evidence of the benefits of psychoactive medications for menopausal symptoms is established, the choice between using hormonal and nonhormonal therapies for management of menopausal symptoms will continue to evolve. PMID- 14563099 TI - Gender differences in depression and response to antidepressant treatment. AB - Throughout their childbearing years, women are twice as likely as men to experience an episode of major depression. The exact etiology of this difference is unclear, but psychosocial and neurobiologic factors likely contribute. Clinicians should consider gender differences in the phenomenology of depression and response to antidepressant treatment when screening for depressive illness, selecting appropriate treatment and assessing therapeutic response. Treatment considerations when selecting an antidepressant should include the patient's gender, age, and, in women, menopausal status, including the use of concomitant HRT. Further research is needed to refine and extend the existing knowledge base regarding the effect of gender on treatment of depression, including the role of endogenous and exogenous gonadal hormones in response to antidepressant treatment. PMID- 14563101 TI - Anxiety disorders in women. AB - Women have higher overall prevalence rates for anxiety disorders than men. Women are also much more likely than men to meet lifetime criteria for each of the specific anxiety disorders: generalized anxiety disorder (GAD), obsessive compulsive disorder (OCD), social anxiety disorder (SAD), posttraumatic stress disorder (PTSD), simple phobia, panic disorder, and agoraphobia. Considerable evidence suggests that anxiety disorders remain underrecognized and undertreated despite their association with increased morbidity and severe functional impairment. Increasing evidence suggests that the onset, presentation, clinical course, and treatment response of anxiety disorders in women are often distinct from that associated with men. In addition, female reproductive hormone cycle events appear to have a significant influence on anxiety disorder onset, course, and risk of comorbid conditions throughout a woman's life. Further investigations concerning the unique features present in women with anxiety disorders are needed and may represent the best strategy to increase identification and optimize treatment interventions for women afflicted with these long-neglected psychiatric disorders. PMID- 14563100 TI - Gender differences in bipolar disorder. AB - The presentation and course of bipolar disorder differs between women and men. The onset of bipolar disorder tends to occur later in women than men, and women more often have a seasonal pattern of the mood disturbance. Women experience depressive episodes, mixed mania, and rapid cycling more often than men. Bipolar II disorder, which is predominated by depressive episodes, also appears to be more common in women than men. Comorbidity of medical and psychiatric disorders is more common in women than men and adversely affects recovery from bipolar disorder more often in women. Comorbidity, particularly thyroid disease, migraine, obesity, and anxiety disorders occur more frequently in women than men, whereas substance use disorders are more common in men. Although the course and clinical features of bipolar disorder differ between women and men, there is no evidence that gender affects treatment response to mood stabilizers. However, women may be more susceptible to delayed diagnosis and treatment. Treatment of women during pregnancy and lactation is challenging because available mood stabilizers pose potential risks to the developing fetus and infant. Pregnancy neither protects nor exacerbates bipolar disorder, and many women require continuation of medication during the pregnancy. The postpartum period is a time of high risk for onset and recurrence of bipolar disorder in women, and prophylaxis with mood stabilizers might be needed. Individualized risk/benefit assessments of pregnant and postpartum women with bipolar disorder are required to promote the health of the woman and avoid or limit exposure of the fetus or infant to potential adverse effects of medication. PMID- 14563102 TI - Sexual function and dysfunction in women. AB - Multiple factors may affect sexual functioning in women, requiring a thorough assessment of all possible etiologies to guide appropriate treatment. Interventions may also be multifaceted, ranging from sex education to psychotherapy to medical treatment. Restoration of sexual functioning is the goal of treatment, but more research is needed for true success to be realized. PMID- 14563103 TI - Psychiatric aspects of endocrine disorders in women. AB - Endocrine disorders are associated with significant psychiatric morbidity. Psychiatric symptoms may manifest as discreet psychiatric syndromes or, more commonly, with a heterogenous group of nonspecific symptoms. Research is needed to understand the relations between hormone excess or deficiency and alterations of mood, cognition, and perception. It is important for clinicians to carefully screen for psychiatric morbidity in patients who present with endocrine disorders. Moreover, patients who appear to present with primary psychiatric syndromes should be carefully evaluated by physical examination and biochemical screening when appropriate to exclude the presence of an underlying endocrine disorder. PMID- 14563104 TI - Mental health issues associated with cardiovascular disease in women. AB - Cardiovascular disease is the leading cause of death in women and men. However, cardiovascular disease and its treatment affect women differently. Much data is emerging regarding the role that mood and anxiety disorders play in the development and prognosis of cardiovascular disease. Pharmacologic treatment of psychiatric disorders can also have direct cardiac effects. The good news is that many more studies are exploring this. However, further research is clearly needed, especially in the area of reproductive hormones and their impact on cardiac disease. PMID- 14563106 TI - Chronic episodic disorders in women. AB - Chronic episodic disorders, such as depressive disorders, IBS, migraine, and FMS, have important commonalities, including cormorbidities, an absence of classic anatomic pathology in the tissues, a lack of objective findings on physical examination, and a lack of abnormal findings by routine laboratory and radiologic tests. These CED are more prevalent in women (perhaps due to changes in estrogen levels), are generally worsened by stress (with resultant hyperactivity of the HPA axis), and often improve with aerobic exercise and common classes of medications affecting serotonin function, such as antidepressants. Thus, an increased understanding of the CED may result in improved treatment and functioning of many patients. PMID- 14563105 TI - Psychiatric aspects of women with breast cancer. AB - Women with breast cancer face not only cancer-related taboos but also issues related to changes in sexuality, femininity, and fertility. Although increased emphasis on the woman's participation in the medical decision-making process has afforded her more treatment options, it also gives her more responsibility in determining her course of treatment through the continuum of cancer care. Patients must deal with a variety of issues related to social reintegration, long term effects of cancer treatment, employment, disability, sexuality, family, and social networks. Screening for distress related to psychological, spiritual/religious, or social concerns can provide guidance for service delivery to those in need. PMID- 14563107 TI - Psychosocial aspects of women's lives: work, family, and life cycle issues. AB - Over the past century and continuing to evolve into the twenty-first century, there have been dramatic changes in work and personal/family lives within the United States. These changes, though strongly affecting men and children, have impacted most dramatically on women's lives, particularly white, middle-class women. Psychiatrists and other mental health clinicians need to be aware of the scope and nature of these changes and to recognize that their own personal experiences and values might differ from those of women of different generations as well as different socioeconomic and cultural backgrounds. PMID- 14563108 TI - Trauma and women. AB - Trauma is extremely prevalent in the lives of women, and the emotional, physical, social, and financial costs are often enduring and pervasive. Given its prevalence, all mental health professionals should receive training in the impact of trauma and learn the principles of its treatment. Although a considerable amount is known about the impact of trauma, much less is known about how to help survivors fully recover, especially women suffering from chronic, complex PTSD. Treatments must-be sensitive to issues of gender. Much research is needed, especially controlled, prospective studies to illuminate normal and pathological responses to trauma. Also needed are large epidemiological studies that compare the genders across trauma types and assess contextual issues of the trauma. We need to learn much more about the individual, family, and community resources that impart resilience, as well as impede recovery. Given the prevalence of violence, it will be impossible to resolve its impact on an individual basis. Interventions also need to be directed at system-level factors that foster violence. PMID- 14563109 TI - Public health context of women's mental health research. AB - As more attention is directed to the mental health care of women, sex and gender differences in research design and in regulatory policies have interfaced with clinical care and public policy. An emphasis on women's mental health issues in the provision of treatment and care as well as the design of large-scale screening strategies to identify and treat women with mental disorders promises to be effective public health approaches to reducing the burden of mental illness in women. The past decade has seen increased emphasis on women's mental health and sex/gender differences in the federal sector and in the research community. Federal regulations (summarized in the NIH Outreach Notebook) call for the inclusion of women and minorities in NIH-funded clinical research. The regulations also place emphasis on gender analysis of the results of clinical trials, in particular phase III trials, the findings of which are likely to influence practice. There has been substantial progress toward the goal of including women in research, but more remains to be done. A 2000 GAO report titled "Women's Health: NIH Has Increased Its Efforts to Include Women in Research" commended NIH for tracking the number of women in clinical research but the report also noted that relatively few NIH-funded studies, including major clinical trials, had reported findings by gender of study participants. This was seen as an impediment to progress in developing gender-based effective treatments. In the past decade, the women's health field has moved beyond an exclusive emphasis on women's reproductive function to one that defines health as a scientific enterprise to identify clinically important sex and gender differences in prevalence, etiology, course, and treatment of illnesses affecting men and women in the population as well as conditions specific to women. Nonetheless, for mental disorders, women's reproductive function and its impact on mental health conditions is still understudied. Based on the epidemiology of mental disorders, the course of mental disorders in women in relation to reproductive transitions remains an important issue for the mental health field because the burden of mental disorders, such as depression and anxiety, fall disproportionately on women of childbearing and childrearing age. The public health emphasis on women's mental health does not lessen the basic scientific opportunities to be had by a focus on gender and sex differences. A 2001 report of the Institute of Medicine titled "Exploring the Biological Contributions to Health: Does Sex Matter?" underscores the benefit to health care of looking for sex differences at the biological level. Basic and clinical neuroscience research is rapidly accruing a knowledge base that will provide information at the level of genes and cells of the influences of biological sex on mental health outcomes in both women and men. A focus on women's mental health and gender/sex differences research promises to yield improvement in treatments and services and thereby to improve the public health as well as to increase fundamental knowledge about the etiology and neurophysiology of mental disorders. PMID- 14563110 TI - Introduction: apoptosis in the development and function of the immune system. PMID- 14563111 TI - Cell death during lymphocyte development and activation. AB - The development and homeostasis of the immune system requires an exquisite balance between cell proliferation and cell death. In this review, we discuss several in vivo and in vitro models that have been developed to help understand the importance of apoptosis during B and T cell development and activation. PMID- 14563112 TI - Cell death in the thymus--it' s all a matter of contacts. AB - Apoptosis, or programmed cell death, plays a critical role in shaping the T cell repertoire, deleting unproductive as well as potentially autoreactive T cells. Our understanding of how thymocyte apoptosis is regulated is continually evolving, as new essential modulators of this process are discovered. A conundrum that remains, however, is how signaling through essentially the same receptors and cascades evokes distinct biological responses: death by neglect, positive or negative selection. We hypothesize that the immunological synapse (IS) may be critical to transducing survival signals during thymocyte development, and suggest that factors affecting IS assembly may also influence T cell selection. PMID- 14563113 TI - Function and regulation of the CD95 (APO-1/Fas) ligand in the immune system. AB - CD95/CD95L mediated apoptosis is an important mechanism of immune homeostasis. It is instrumental for termination of an immune response and mainly be involved in peripheral tolerance. Dysregulation of the CD95/CD95L system leads to severe diseases. In this review, we present a survey of the role of the CD95/CD95L system in the immune system and, particularly, focus on the signals and transcription factors (NF-AT, Egr, NF-kappaB, AP-1, c-Myc, Nur77, IRFs, SP-1, ALG 4, ROR(gamma)t, and CIITA) involved in CD95L expression. It should also be evident from this review that a profound insight into the molecular mechanisms of CD95L activation should allow to explore potential therapeutic means to treat CD95/CD95L-dependent diseases. PMID- 14563114 TI - The role of Rel/NF-kappaB transcription factors in B lymphocyte survival. AB - Among the transcriptional mediators of the various signal transduction pathways that have been shown to regulate lymphocyte apoptosis, Rel/NF-kappaB transcription factors have emerged as key regulators of B cell survival during their differentiation and in their activation by antigens or mitogens. The aim of this review is to bring together recent findings on Rel/NF-kappaB regulation of B cell survival and to present an integrated model of how these transcription factors control apoptosis in a signal and differentiation-stage specific manner. In addition to providing a contemporary view of Rel/NF-kappaB regulated B cell survival, the model described here is aimed to serve as a paradigm for how Rel/NF kappaB family members control survival in different cell types during differentiation and in response to the plethora of signals that impinge on this master transcriptional regulatory pathway. PMID- 14563115 TI - Fas (CD95/Apo-1) ligand regulation in T cell homeostasis, cell-mediated cytotoxicity and immune pathology. AB - Members of the tumor necrosis factor (TNF) superfamily are crucially involved in the regulation of T cell activation, homeostasis and cytotoxicity. In particular, Fas ligand (FasL), expressed by activated T lymphocytes, induces cell-mediated cytotoxicity and may also be responsible for apoptotic suicide. Tight regulation of this death-inducing ligand is a prerequisite for proper immune defense and homeostasis. In this review, we will discuss various aspects of FasL regulation in cell-mediated cytotoxicity, immune homeostasis and the immunopathology of diseases. PMID- 14563116 TI - From antigen to activation: specific signal transduction pathways linking antigen receptors to NF-kappaB. AB - A precise balance between cellular apoptosis and cellular survival is essential for the proper functioning of the immune system. Whereas apoptosis eliminates self-reactive or supernumerary lymphocytes, survival signaling that counteracts apoptotic programs is needed to allow B and T lymphocytes that recognize pathogens to become activated and expand in response to infection. A major regulator of lymphocyte survival and activation is the transcription factor NF kappaB. Controlled activation of NF-kappaB is essential for normal innate and adaptive immune responses, and dysregulated NF-kappaB signaling in lymphocytes contributes to diseases ranging from chronic inflammation and autoimmunity to lymphoma. The core NF-kappaB activating machinery composed of the NF-kappaB, IkappaB and IKK proteins is relatively well-characterized, but it is less clear how distinct upstream stimuli activate NF-kappaB in a tissue-, time- and signal specific manner. In this review, we discuss recent insights into the specific signal transduction pathways leading to NF-kappaB activation that are triggered by engagement of the antigen receptors of T and B cells. We focus mainly on T cell receptor (TCR)-mediated NF-kappaB activation and draw parallels to B cell receptor (BCR)-mediated NF-kappaB activation where appropriate. PMID- 14563117 TI - The CD95 type I/type II model. AB - CD95 (APO-1/Fas) has become the prototype of a death domain containing receptor and is the best studied member of the death receptors that activate the extrinsic apoptosis pathway. This pathway is initiated by recruitment and activation of caspase-8, an initiator caspase, in the death-inducing signaling complex (DISC) followed by direct cleavage of downstream effector caspases. In contrast, the intrinsic apoptosis pathway starts from within the cell either by direct activation of caspases or through intracellular changes such as DNA damage resulting in the release of a number of pro-apoptotic factors from the intermembrane space of mitochondria. The release of these factors results in the activation of another initiator caspase, caspase-9, and ultimately in the activation of effector caspases in a protein complex called the apoptosome. In recent years, it has become apparent that there is cross talk between the extrinsic and intrinsic pathway. In the death receptor pathway of apoptosis induction, the best characterized connection between the two pathways is the Bcl 2 family member Bid which translocates to mitochondria after cleavage by caspase 8 causing pro-apoptotic changes. Cells that die through CD95 without help from mitochondria are called Type I cells, whereas cells in which CD95-mediated death relies mostly on the intrinsic pathway are called Type II. This review focuses on recent developments in the delineation of the biochemistry and the physiological function of the two CD95 pathways. PMID- 14563118 TI - SHP-1: a regulator of neutrophil apoptosis. AB - The Src homology domain 2 (SH2)-containing tyrosine phosphatase-1 (SHP-1) has been implicated in the regulation of differentiation, proliferation, and activation of hematopoietic cells. In this review, we discuss the potential role of SHP-1 in modulating apoptosis pathways in neutrophils. Low enzymatic SHP-1 was associated with increased neutrophil survival in vitro and SHP-1-deficient mice were reported to develop severe neutrophilic inflammatory responses. In contrast, high expression of this phosphatase was observed in neutropenic patients. Moreover, when neutrophils were exposed to Fas ligand, TNF-alpha, or TRAIL, the anti-apoptotic effects of granulocyte-macrophage colony-stimulating factor (GM CSF), granulocyte colony-stimulating factor (G-CSF), or IFN-gamma were blocked, most likely by SHP-1-mediated inactivation of anti-apoptotic signaling molecules. In summary, the current available data point to an important role of SHP-1 in the regulation of neutrophil apoptosis. PMID- 14563119 TI - MicroRNAs and cancer. AB - The general basis of cancer is the loss of cell identity and inappropriate proliferation of cells. Classically, a universal paradigm in oncogenesis is the accumulation of mutations in the open reading frames of protein-encoding oncogenes and tumor suppressors. The identification of new classes of noncoding RNAs (ncRNA) important for development and cell homeostasis will likely change this current paradigm. Recent data suggests that a special class of ncRNAs called microRNAs might be involved in human disease. This review proposes a role for microRNAs in oncogenesis. PMID- 14563120 TI - RNA interference: gene silencing in the fast lane. AB - Sequencing of whole genomes has provided new perspectives into the blueprints of diverse organisms. Knowing the sequences, however, does not always tell us much about the function of the genes that regulate development and homeostasis. RNA interference (RNAi) is becoming the method of choice for gene function analysis in cells and whole organisms. Here we review the approaches available to perform RNAi experiments in mammalian cells and in mice. We discuss usage of RNAi in cancer research and as a possible therapeutic tool for cancer treatment. PMID- 14563121 TI - Ras interference as cancer therapy. AB - Activating point mutations of the small GTPase Ras are present in about 30% of all human tumors. Constitutively active Ras induces growth factor independent cell proliferation and cell survival. Oncogenic Ras appears to be essential for tumor progression and maintenance. Several therapeutic agents have been developed to inhibit Ras, such as FTIs and antisense oligonucleotides. A new tool for blocking oncogenes in cancer cells has emerged with the discovery that RNA interference can specifically silence expression of endogenous human genes. The therapeutic potential of a RNAi-mediating vector was recently demonstrated by the stable suppression of oncogenic K-Ras in tumor cells. PMID- 14563122 TI - Potentials for RNAi in sarcoma research and therapy: Ewing's sarcoma as a model. AB - Existing data identify EWS-FLI1 as indispensable for sustained Ewing's sarcoma growth and as the ideal therapeutic target in this disease. The siRNA may hold great promises as a fusion gene specific agent. RNAi mediated suppression of EWS FLI1 is likely to result in an altered tumor cell phenotype including changes in chemosensitivity, and a restored differentiation potential. Thus, RNAi may serve as an adjuvant to chemotherapy. As a therapeutic means however, RNAi is hampered by limitations in the delivery of the agent and emergence of resistant clones. In vitro suppression of EWS-FLI1 expression will allow to define the phenotypic characteristics of dormant tumor cells that may give rise to late relapses, enabling improved diagnosis and treatment even of minimal residual disease. PMID- 14563123 TI - Targeting oncogenic fusion genes in leukemias and lymphomas by RNA interference. AB - Leukemias and lymphomas are often characterized by non-random chromosomal translocations that, at the molecular level, induce the activation of specific oncogenes or create novel chimeric genes. They have frequently been regarded as optimal targets for gene-silencing approaches because of the large body of evidence that these single abnormalities directly initiate and maintain the malignant process. Herein, we discuss RNA interference (RNAi)-based approaches for targeting the fusion sites of chromosomal translocations as a future treatment option in leukemias and lymphomas. PMID- 14563125 TI - Applications of RNA interference for studies in DNA damage processing, genome stability, mutagenesis, and cancer. AB - RNA interference, discovered only five years ago, is an important system for the control of gene expression. It is also quickly becoming a valuable experimental tool as it allows investigators to knock down the level of expression of specific genes. In this paper, we review some applications of this technology for studies in DNA damage processing, genome stability, mutagenesis, and cancer. PMID- 14563124 TI - Exposing oncogenic dependencies for cancer drug target discovery and validation using RNAi. AB - Oncogenesis occurs through the acquisition and selection of multiple somatic mutations--each contributing to the growth, survival and spread of the cancer. Key attributes of the malignant phenotype, such as unchecked proliferation and cell survival, can often be "reversed" by the selective diminution of dominant oncogenes by chemical or genetic means (e.g. beta-catenin in colorectal carcinomas; bcr-abl in chronic myelogenous leukemias (CMLs)). These observations suggest that the products of oncogenes, or of secondary genes that mediate and maintain tumor phenotypes, might be revealed through the systematic disruption of each and every gene in tumor-derived cells. Some of these genes may encode proteins amenable to therapeutic intervention, thus fueling the cancer drug discovery process. However, a functional assessment of each known or predicted gene in mammalian cells is a daunting task and represents the rate-limiting step in drug target identification and validation. In this regard, RNA interference (RNAi) by small interfering RNAs (siRNA) holds great promise as the "tool of choice" to mediate the selective attenuation of mammalian gene expression and protein function. Here, we review strategies by which RNAi might be used to determine the genetic alterations that contribute to malignant transformation via large-scale cell-based screens, and propose how this information can be used in conjunction with small molecule screens to identify pathways critical to cancer cell survival. PMID- 14563126 TI - Tumor specific activation of PKR as a non-toxic modality of cancer treatment. AB - Over the past decade progress has been made in the development of therapies against cancer. Small molecules, mainly tyrosine kinase inhibitors (tyrphostins) like Gleevec, Iressa targeting CML and EGFR overexpressing tumors have entered the clinic, where a large number of other tyrphostins are at various stages of clinical development. In parallel a few antibodies like Herceptin targeting breast cancer overexpressing Her-2 and Rituxan targeting B cell malignancies are utilized in the clinic. In all these cases success is moderate and restricted to a narrow population of patients, except for Gleevec which is effective for a long duration for chronic CML. The cancer community agrees that this is actually a unique exception that proves the rule. Over the past few years a few modalities of cancer gene therapies have emerged. In this short review we shall summarize our efforts to develop methods to activate PKR selectively in cancer cells. PMID- 14563127 TI - Dynamic changes of rDNA condensation state during mitosis in filamentous fungi revealed by fluorescence in situ hybridisation. AB - We visualised rDNA of two ascomycetes, Cochliobolus heterostrophus and Haematonectria haematococca, by fluorescence in situ hybridisation (FISH) and analysed the condensation state of rDNA during mitosis. Both fungi showed a similar course of change in rDNA condensation corresponding to different mitotic stages. rDNA was decondensed in its entire length at interphase, and became increasingly condensed during prophase. The condensation reached a maximum at metaphase, remaining in that state through anaphase. Metaphase observation revealed the single distal location on a chromosome of rDNA in each fungus. This study provides the first visual evidence of the cyclic change of the condensation/decondensation state of rDNA during mitosis in filamentous fungi by FISH. PMID- 14563128 TI - Morphology and ultrastructure of Neolecta species. AB - Several independent molecular phylogenetic analyses have indicated that the genus Neolecta has a unique position within the Ascomycota. It is the only taxon outside the core-group of filamentous, ascoma-forming ascomycetes that also has the ability to form ascomata. Light and electron microscope studies indicate that hymenial structure and development in Neolecta spp. are unique. Ascogenous hyphae in N. vitellina branched repeatedly and successively to produce asci. Non ascogenous hyphae were multinucleate, often with nuclei in pairs. Nuclear pairing was particularily prominent in the ascogenous hyphae. A basal septum delimited the dikaryotic ascus. Ascosporogenesis was initiated by nuclear fusion followed by a meiotic and mitotic division to form eight nuclei. The ascus apex was thin with an annular subapical thickening. Ascospores were forcibly released through a 'split' in the ascus apex. Woronin bodies were frequently associated with hyphal septa. Attempts to culture N. vitellina and to obtain molecular information from the type species, N. flarovirescens, were unsuccessful. However, N. flavovirescens showed several microscopic characters that indicated close relationships with the two other species in the genus, N. vitellina and N. irregularis. The position of Neolecta spp. within the Ascomycota is discussed. PMID- 14563129 TI - Phylogenetic relationships of the genus Phanerochaete inferred from the internal transcribed spacer region. AB - Phanerochaete is a genus of resupinate homobasidiomycetes that are saprophytic on woody debris and logs. Morphological studies in the past indicated that Phanerochaete is a heterogeneous assemblage of species. In this study the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA was used to test the monophyly of the genus Phanerocthaete and to infer phylogenetic relationships of the 24 taxa studied. Maximum parsimony, maximum likelihood, and Bayesian analyses do not support the monophyly of the genus. However, a core group of species represented by Phanerochaete velutina, P. chrysosporium, P. sordida, P. sanguinea and others are closely related and group together in a clade. Other common Phanerochaete species including Phanerochaete rimosa, P. chrysorhiza, P. omnivora, P. avellanea, P. tiberculata, P. flava, and P. allantospora, however, do not cluster with the core Phanerochaete group. PMID- 14563130 TI - Identification and molecular phylogeny of Epulorhiza isolates from tropical orchids. AB - 24 isolates of Epulorhiza were obtained from the roots and protocorms of orchids in Singapore. Two groups were distinguished based on morphological and cultural characteristics. Group 1 comprised 20 isolates which were identified as E. repens, while those in group 2 were identified as E. calendulina-like Rhizoctonia. The ITS-5.8S rDNA sequence identity ranged from 88-100% among isolates of E. repens and six sub-groups were further delineated. The sequence identity was 98-100% among E. calendulina-like Rhizoctonia isolates. The sequence identity between E. repens isolates and E. calendulina-like Rhizoctonia isolates ranged from 18-44%. Apparently, isolates from both groups 1 and 2 were genetically distinct. Phylogenetic analysis showed that the distribution of the isolates correlated with the sites and the hosts from which the fungi were isolated. No matching sequences have been found in the GenBank database for the ITS region of E. repens and E. calendulina-like Rhizoctonia. PMID- 14563131 TI - Determination of biological species and analysis of genetic variability by RAPD of isolates of Pleurotus subgenus Coremiopleurotus. AB - Isolates of Pleurotus cystidiosus and P. smithii were studied to verifie the occurrence of P. cystidiosus instead of P. smithii in South America. The two species are mainly separated by the growth rate of the anamorph in culture, the morphology of the anamorph and teleomorph, intercompatibility tests, and genetic variability. In order to see if the isolate found in Brazil and previously identified as Antromycopsis macrocarpa (the anamorph of P. cystidiosus) belongs to P. cystidiosus, a species with a world wide distribution, or to P. smithii which is restricted to Mexico and South America, or if P. cyistidiosus and P. smithii are the same species, isolates of different geographic origins were studied. Growth rate in culture, mono-dikaryotic matings, and genetic variability determined by RADP were investigated. The results show that the criteria used to separate the two species are unsatisfactory, and that P. smithii should be considered a synonym of P. cystidiosus; this extends the distribution of this later species to Central and South America. PMID- 14563132 TI - Telomeric fingerprinting of the violet root rot fungus, Helicobasidium mompa: a useful tool for karyotype estimation. AB - We hybridized the telomere-associated DNA sequence pTel46 isolated from Coprinus cinereus with Helicobasidium mompa genomic DNA. The hybridized fragments were more sensitive to Bal31 nuclease than those that were not hybridized, suggesting that they were located at the ends of chromosomes in H. mompa. The hybridization profile can be used to estimate chromosome number, since the number of chromosomes in a single basidiospore isolate is about half that in putative parent strains. Thus, single basidiospore and field isolates might be homokaryons and heterokaryons respectively. We found telomere-linked restriction fragment length polymorphisms (RFLPs) in strains of H. mompa isolated from field and individual basidiosporcs. Thus, this marker appears to be an excellent tool with which to reveal the considerable polymorphism of H. mompa and to identify strains. The RFLP was not found among several strains of the same mycelial compatibility group (MCG) isolated from the same field, suggesting that strains belonging to an MCG group are identical. PMID- 14563133 TI - Heterologous diploid nuclei in the violet root rot fungus, Helicobasidium mompa. AB - Allelic genes hga1-1 and hga1-2, which encode G protein alpha subunit in the violet root rot fungus, Helicobasidium mompa, were sequenced and characterized. Restriction fragment polymorphism (RFLP) analysis determined that the gene is present as a single locus in the single basidiospore isolates, while strain V169 possessed both alleles of this gene. Therefore, although basidiospore isolates are dikaryon, they are homokaryotic. Field-isolated strain V169, the putative parent strain, is a dikaryotic heterokaryon. Allelic genes hga1-1 and hga1-2 segregated in almost a 1:3 ratio among single basidiospore isolates from the same fruiting body. Moreover, the copy number of hga1-1 was found to be less than that of hga1-2 in the V169 strain. These results suggest that one of the nuclei in the V169 parent strain is homozygous diploid and the other heterozygous diploid. This parent strain produced four homokaryotic and dikaryon basidiospores on each basidium. PMID- 14563134 TI - RAPD analyses of recombination processes in the entomopathogenic fungus Beauveria bassiana. AB - To understand the nature of recombination processes in Beauveria bassiana, double auxotrophic complementary mutant strains were used to produce six heterokaryons by three different methods. Conidia from these heterokaryons were plated on selective media and stable haploid (but not diploid) recombinants were isolated. Single colony recombinants were recovered with both parental and non-parental random amplified polymorphic DNA (RAPD) profiles. These results suggest that a range of different recombination mechanisms may be occurring in B. bassiana. PMID- 14563135 TI - Action spectrum for the induction of hyphal branches of an arbuscular mycorrhizal fungus: exposure sites versus branching sites. AB - The first action spectrum for a photo-induced response of an arbuscular mycorrhizal fungus is reported. At low light intensity, the responsive wavelengths for light-induced hyphal branching of the primary germ tube of Gigaspora gigantea were determined to be in the blue to uv-A range. The action spectrum showed the greatest stimulation of branching occurred around 390 nm although a shoulder was observed between 360-370 nm. A second major peak of light induced branching occurred at 430 nm. The exposure of specific areas of the germ tube to high intensity blue light for a short period led to several interesting observations. By exposing 2 mm segments (0-2, 2-4, 4-6, etc.) or 3 mm segments away from the tip, it was determined that photoinduction of hyphal branches could occur anywhere along the axis of a growing germ tube except in the apical 2 mm. When 3 mm segments were exposed at greater distances from the tip (6-9, 9-12, and up to 33-36 mm), branches frequently formed in areas not directly exposed to light. The branches were usually in clusters which were spaced approximately 3 or 6 mm apart. Since light scattering could be ruled out, these results indicated that the exposure sites and sites of hyphal branching did not necessarily coincide and suggested the probable involvement of a second messenger during this blue light-induced event. PMID- 14563136 TI - Differential expression of Glomus intraradices genes in external mycelium and mycorrhizal roots of tomato and barley. AB - Relative quantitative RT-PCR and western blotting were used to investigate the expression of three genes with potentially regulatory functions from the arbuscular mycorrhizal fungus Glomus intraradices in symbiosis with tomato and barley. Standardisation of total RNA per sample and determination of different ratios of plant and fungal RNA in roots as colonisation proceeded were achieved by relative quantitative RT-PCR using universal (NS1/NS21) and organism-specific rRNA primers. In addition, generic primers were designed for amplification of plant or fungal beta-tubulin genes and for plant glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes as these have been suggested as useful controls in symbiotic systems. The fungal genes Ginmyc1 and Ginhb1 were expressed only in the external mycelium and not in colonised roots at both mRNA and protein levels, with the proteins detected almost exclusively in the insoluble fractions. In contrast, mRNA of Ginmyc2 was identified in both external and intraradical mycelium. In mycorrhizal roots, Ginmyc2 and fungal beta-tubulin mRNAs increased in proportion to fungal rRNA as colonisation proceeded, suggesting that accumulation reflected intraradical fungal growth. Fungal alpha-tubulin protein and beta-tubulin mRNA both appeared to be more abundantly accumulated in AM hyphae within heavily colonised roots than in external hyphae, relative to fungal rRNA. Tomato GAPDH mRNA accumulation was proportional to tomato rRNA, but accumulation of tomato beta-tubulin mRNA was reduced in colonised roots compared to non-mycorrhizal roots. These results provide novel evidence of differential spatial and temporal regulation of AM fungal genes, indicate that the expression of tubulin genes of both plant and fungus may be regulated during colonisation and validate the use of multiple 'control' genes in analysis of mycorrhizal gene expression. PMID- 14563137 TI - Infection of barley roots by Chaetomium globosum: evidence for a protective role of the exodermis. AB - Plant pathogenesis by fungi is known to be dependent on the host genotype, the virulence of the pathogen and certain environmental conditions influencing fungal establishment. Previously, it has been shown that Chaetomium globosum, a fungus well-characterized for its biocontrol potential, causes necrosis on barley roots grown in Murashige and Skoog (MS)-agar. Using MS-agar and aeroponic culture as axenic plant growth systems, C. globosum pathogenesis was analyzed with serological and histological methods. Irrespective of the growth system, C. globosum infected the root epidermis. Roots grown in MS-agar were extensively colonized intercellularly and intracellularly up to the inner cortex and the tissue underwent necrosis. In contrast, roots grown in aeroponic culture were not colonized beyond the epidermis and the roots appeared healthy. Histochemical analyses revealed that hypodermal suberization stopped fungal invasion. In root tips known to lack suberization, epidermal papilla formation reduced overall infection frequency. The results indicate that specific environmental conditions are important for infection and disease expression in barley roots. Infection is restricted by two spatial and temporal distinct defence mechanisms: (1) papillae in root tips retarding fungal invasion; and (2) suberization of hypodermal cells blocking fungal radial growth. PMID- 14563138 TI - Responses of the root rot fungus Collybia fusipes to soil waterlogging and oxygen availability. AB - Collybia fusipes is a common root rot fungus in mature pedunculate oak forest, that causes drastic destruction of the tree root systems, especially in dry or mildly waterlogged soils. We wanted to check, under controlled conditions or in forest ecosystems, whether reduced O2 during saturation of the soil by water could interact with disease evolution. Susceptibility of waterlogged oak seedlings to C. fusipes was tested in a greenhouse and the survival of the pathogen in woody substrates was assessed in hydromorphic soils in a forest. A direct and detrimental effect of soil waterlogging on C. fusipes survival was evidenced both under controlled conditions and in forest stands. Growth of C. fusipes mycelium on agar media was monitored under low O2 mole fraction and compared to that of Armillaria mellea and Heterobasidion annosum. A drastic reduction in mycelial growth was evidenced in C. fusipes and H. annosum but not in A. mellea. PMID- 14563139 TI - Ascobotryozyma cognata sp. nov., a new ascomycetous yeast associated with nematodes from wood-boring beetle galleries. AB - A new species of Ascobotryozyma, A. cognata sp. nov. (anamorph Botryozyma cognata), was isolated from beetle galleries in Idaho, USA. A. cognata was found on the surface of free-living nematodes, Panagrellus dubius, collected from galleries created by the long-horned beetle Saperda calcarata in Populus (aspen), and the weevil Cryptorhynchus lapathi in Salix (willow). A. cognata isolates were collected from similar habits and in relatively close proximity to those of A. americana, the only species described from North America. The recognition of A. cognata as a distinct species was supported by morphological and molecular data. Thallus cells of A. cognata were significantly shorter than those of A. americana. Low DNA reassociation values, notably different randomly amplified polymorphic DNA (RAPD), inter-sequence simple repeat (ISSR), and amplified fragment-length polymorphic (AFLP) fingerprints, and sequence divergence in both the D1/D2 domain of the nuc-LSU rDNA and an additional unidentified region were all consistent with the recognition of a new species. PMID- 14563140 TI - One-level one-sided lumbar disc surgery with and without microscopic assistance: 1-year outcome in 114 consecutive patients. AB - OBJECT: The aim of this study was to compare the outcomes following macrodiscectomy and microsurgery for one-level one-sided lumbar disc excision. METHODS: The authors prospectively studied 114 consecutive patients who underwent microdiscectomy (Group A, 63 patients [36 men, 27 women]) and macrodiscectomy (Group B, 51 patients [29 men, 22 women]) for one-level unilateral first-time lumbar disc herniation. Microdiscectomy was considered to involve a small incision with removal or opening of the ligamentum flavum, no or minimal bone excision, and use of the operating microscope to remove the disc material. Laminectomy combined with macrodiscectomy was defined as any operation requiring a large opening in or complete removal of the unilateral lamina. Diagnosis was confirmed by magnetic resonance imaging. A 1-year follow-up investigation was also conducted. Relief of radicular pain, improvement in muscle power, and changes in sensory and/or reflex abnormality were documented. Assessment of outcome was performed using the modified Stauffer-Coventry criteria. Good or excellent results were demonstrated in 90% of Group A and 89% of Group B patients (p > 0.05). One patient in each group underwent reoperation. There was infection over the fascia in two Group A patients. Mean operative time (+/- standard deviation) was 54 +/- 5.65 minutes in Group A and 25 +/- 7.07 minutes in Group B (p < 0.01). Median length of the incision was 4 and 6 cm in Group A and Group B, respectively (p < 0.05). The length of postoperative inpatient stay was 1 day in both groups (p > 0.05). Patients in the microsurgery-treated group returned to work in less time: 85% of Group A and 58% of Group B patients returned to their work within 4 weeks (p < 0.001). Some patients in each group (15% in Group A and 45% in Group B) needed narcotic analgesic medication at least twice between the 1st month and 1st year after the surgery (p < 0.001). CONCLUSIONS: Microdiscectomy allows the surgeon good visualization and is less traumatic to the involved tissues. Interestingly, the results of this study indicated that microsurgery does not reduce hospitalization time, nor does it improve the overall surgery-related outcome. The main differences between the two procedures were length of the incision and operative time. The author found that lumbar microdiscectomy allows patients earlier return to work and/or normal life with less reliance on postoperative narcotic analgesic agents. PMID- 14563141 TI - Factors related to long-term outcome after decompressive surgery for ossification of the ligamentum flavum of the thoracic spine. AB - OBJECT: Factors related to long-term surgical outcome of thoracic myelopathy caused by ossification of the ligamentum flavum (OLF) have not been fully investigated. To evaluate these factors, the authors reviewed medical records obtained in patients who had undergone decompressive surgery for thoracic OLF. METHODS: Thirty-four patients in whom decompressive surgery was performed for thoracic OLF (mean follow-up period 8 years) were recruited. Fourteen patients underwent simultaneous decompressive surgery for cervical or lumbar lesions. Patient age, duration of symptoms, OLF type, dural adhesion, neurological status classified by the Japanese Orthopaedic Association (JOA) scale before and after surgery (1 month postoperatively and at final follow up), and recovery rate were reviewed. The ossified ligamentum flavum was classified into five types according to Sato's classification by assessing preoperative computerized tomography scans. The mean preoperative JOA score (5) was significantly improved at 1 month after surgery (7.1) and at final follow up (7.9) (p < 0.05). Recovery rates at 1 month and at final follow up were 34.6 and 44.2%, respectively. No significant correlations were found between age, OLF type, and neurological status before and after surgery. In patients with dural adhesion, which was observed in cases of larger-type OLF (p < 0.02), lower JOA scores were observed compared with those in patients without dural adhesion both preoperatively and at 1 month after surgery (p < 0.05) but not at the final follow-up examination. Higher JOA scores and recovery rates were demonstrated in patients who had undergone simultaneous surgery than in those who had not undergone simultaneous surgery at 1 month (p < 0.05) but not at the final follow up. Using multiple regression analysis, the authors identified the duration of preoperative symptoms as the most important predictor of a high JOA score and recovery rate at the final follow up. CONCLUSIONS: Duration of preoperative symptoms represents the most important predictor of long-term surgery-related outcome in patients treated for thoracic OLF. The type of ossified ligamentum flavum, dural adhesion, and simultaneous surgery for coexistent cervical or lumbar lesions do not appear to influence the long-term postoperative prognosis. PMID- 14563142 TI - Improved accuracy of computer-assisted cervical pedicle screw insertion. AB - OBJECT: The authors introduce a unique computer-assisted cervical pedicle screw (CPS) insertion technique used in conjunction with specially modified original pedicle screw insertion instruments. The accuracy of screw placement as well as surgery-related outcome and complication rates were compared between two groups of patients: those in whom a computer-assisted and those in whom a conventional manual insertion technique was used. METHODS: The screw insertion guiding system consisted of a modified awl, probe, tap and a screwdriver specially designed for a computer-assisted CPS insertion. Using this system, real-time instrument/screw tip information was three dimensionally identified in each step of screw insertion. Seventeen patients underwent CPS fixation in which a computer-assisted surgical navigation system was used. The cervical disorders consisted of spondylotic myelopathy with segmental instability or kyphosis, metastatic spinal tumor, rheumatoid spine, and postlaminectomy kyphosis. The rate of pedicle wall perforation was significantly lower in the computer-assisted group than that in the other group (1.2 and 6.7%, respectively; p < 0.05). The screw trajectory in the horizontal plane was significantly closer to the anatomical pedicle axis in the computer-assisted group compared with the manual insertion group (p < 0.05). This factor significantly reduced the incidence of screw perforation laterally. Complications such as neural damage or vascular injury were not demonstrated in the computer-assisted group (compared with 2% in the manual insertion treatment group). The overall surgery-related outcome was satisfactory. CONCLUSIONS: In contrast to the previously reported computer-assisted technique, our CPS insertion technique provides real-time three-dimensional instrument/screw tip information. This serves as a powerful tool for safe and accurate pedicle screw placement in the cervical spine. PMID- 14563143 TI - Clinical and radiological evaluation of the Codman semiconstrained load-sharing anterior cervical plate: prospective multicenter trial and independent blinded evaluation of outcome. AB - OBJECT: Semiconstrained load-sharing implants for spinal fixation accommodate change in the screw-plate interface as bone grafts shrink. The authors evaluated the clinical and radiological outcome in patients after placement of the Codman anterior cervical plate (ACP) system, which allows change in the screw-plate angle. METHODS: The authors undertook a 10-center prospective study with independent blinded evaluation. All patients underwent cervical fusion and placement of ACPs. Clinical and radiological evaluations were performed at 1, 3, 6, 12, and 24 months. Radiographs were examined for screw angles, construct height, fusion, and screw fracture or displacement. One hundred ninety-five patients were enrolled. The mean follow-up period was 17 months. At 24 months neurological improvement was demonstrated in 68.7% and pain improvement in 76.6% of the patients. Fusion was successful in 93.8%. Varying degrees (most minor) of hardware-related failure occurred in 10.4% of cases; however, reoperation was required in only four (2.1%). A significant change in screw angles occurred over time (mean 6.4 degrees in caudal screw angle [p < 0.001] and 2.4 degrees in the rostral screw angle [p = 0.003]). These changes plateaued by 6 months. A change in construct height (mean 3.48 mm) occurred by 6 months (p < 0.05). CONCLUSIONS: Based on an independent blinded evaluation, the Codman ACP provides effective fixation with load sharing and is effective in achieving fusion with a 94% success rate. Direct comparison with rigidly locked devices is required to establish definitively the optimal method for anterior cervical fixation. PMID- 14563145 TI - Mechanism of the spinal cord injury and the cervical spondylotic myelopathy: new approach based on the mechanical features of the spinal cord white and gray matter. AB - OBJECT: The authors have previously investigated the mechanical properties of the white and gray matter in the bovine cervical spinal cord, demonstrating that the gray matter is more rigid, although more fragile, than the white matter. In the present study they conducted additional tensile tests on the bovine cervical spinal cord by changing strain levels and strain rates applied to the white and gray matter. METHODS: Based on their testing, the authors found the following: 1) Stress within the spinal cord relaxes over time. 2) Intracord stress is related to the strain rates or levels. The finite element method was used to compute the stress distribution within the spinal cord under three compressive loading conditions. Results from the computations showed a different stress distribution in the white and gray matter, where the distribution of stress varied with strain rate, compression volume, and the position of compression. CONCLUSIONS: These differences in mechanical properties between the white and gray matter constitute different mechanisms contributing to the development of tissue damage and clinical symptoms. PMID- 14563144 TI - Low-dose radiotherapy for the inhibition of peridural fibrosis after reexploratory nerve root decompression for postlaminectomy syndrome. AB - OBJECT: The authors of clinical studies have demonstrated a significant association between the presence of extensive post-lumbar discectomy peridural scar formation and the recurrence of low-back and radicular pain. Low-dose perioperative radiotherapy has been demonstrated to inhibit peridural fibrosis after laminectomy in animal models. The present study was designed to evaluate the clinical efficacy of preoperative irradiation in patients with failed-back surgery syndrome due to peridural fibrosis who underwent reexploration and nerve root decompression. METHODS: Ten patients with symptomatic post-discectomy peridural fibrosis were randomized. Half of the patients underwent 700-cGy external-beam irradiation to the operative site 24 hours prior to reexploration and decompressive treatment of their symptomatic nerve root(s) (treatment group) and the other half underwent reexploration and decompressive treatment without preoperative irradiation (control group). All patients underwent simulated irradiation so neither patient nor surgeon was aware of the patient's group. In all patients the antiadhesion product ADCON-L was placed over the affected nerve root at the time of surgery. Clinical outcome was assessed using the American Association of Neurological Surgeons/Congress of Neurological Surgeons Joint Section Lumbar Disc Herniation Study Questionnaire at baseline, 6 weeks, 3 months, and 1 year follow up. Five men and five women (mean age 42 years) underwent randomization and surgery. Three patients underwent reexploration at L4 5, four at L5-S1, and three at both levels. No complication was associated with irradiation, and no new neurological deficits occurred. At 1-year follow-up examination, three irradiation-treated patients were pain free and two experienced improvement. In the control group, three patients experienced improved pain relief and two were unchanged. There was a trend toward better outcome at 1 year in the radiotherapy-treated group (p = 0.056). CONCLUSIONS: Preoperative low-dose external-beam irradiation improved clinical outcomes after reexploration and decompression of nerve roots affected by postlaminectomy peridural fibrosis causing radicular pain. The addition of preoperative irradiation may improve outcome in patients who undergo reoperation for recurrent radicular pain associated with a significant amount of peridural fibrosis, particularly now that no antiadhesion product is available for clinical use. PMID- 14563146 TI - Spinal blood flow in 24-hour megadose glucocorticoid treatment in awake pigs. AB - OBJECT: Because of the controversy regarding the benefits of 24-hour administration of methylprednisolone in patients with spinal cord injury (SCI), it is important to investigate its mechanism of action and side effects. This study was conducted to determine if high-dose methylprednisolone modulates neural and vertebral blood flow in an awake large-sized animal model without SCI. METHODS: From a group of 18 immature female domestic pigs born to nine different litters, nine animals were randomly allocated to receive methylprednisolone treatment, whereas their nine female siblings served as controls. Drug or placebo was applied in a blinded fashion by a third person not involved in the study. The following treatment for SCI, as suggested by the North American Spinal Cord Injury Study, was administered to the awake pig: methylprednisolone (30 mg/kg of body weight) was infused into the jugular vein during a 15-minute period, followed by a 45-minute pause, and the infusion was maintained over a 23-hour period at a dose of 5.4 mg/kg body weight/hour. By means of the radioactive tracer microsphere technique, spinal cord blood flow (SCBF) was measured in the awake standing pig in the cerebrum, and in spinal gray and white matter, nerve roots, endplates, cancellous bone, cortical shell, and T12-L2 discs. Blood flow was measured before, 1 hour after initiation of infusion, and 24 hours postinfusion. Examination of blood flow in the neural and vertebral tissue samples, as well as of central hemodynamics, revealed no significant difference between the experimental and control groups, and this parity was maintained throughout the experimental phases. CONCLUSIONS: In the awake pig model, 24-hour methylprednisolone treatment does not modulate cerebral or SCBF, nor does it increase the risk for vertebral osteonecrosis by producing vertebral ischemia. PMID- 14563148 TI - Changes in nerve root motion and intraradicular blood flow during intraoperative femoral nerve stretch test. Report of four cases. AB - OBJECT: It is not known whether changes in intraradicular blood flow (IRBF) occur during the femoral nerve stretch test (FNST) in patients with lumbar disc herniation. An FNST was conducted in patients with lumbar disc herniation to observe the changes in IRBF, and results were then compared with clinical features. METHODS: The study was composed of four patients with L3-4 disc herniation who underwent microdiscectomy. Patients were placed prone immediately before surgery, so that their knee flexed on the operating table with the hip joint kept in hyperextension, and the FNST was performed to confirm at which region pain developed in the anterolateral thigh. During the operation, the hernia-affected nerve roots were visualized under a microscope. The needle sensor of a laser Doppler flowmeter was then inserted into each nerve root immediately above the hernia, and the change in IRBF was measured during the intraoperative FNST. After removal of the herniated disc, a similar procedure was repeated and IRBF was measured again. The intraoperative FNST showed that the hernia compressed the nerve roots and there was marked disturbance of gliding, which was reduced to only a few millimeters. During the test, IRBF decreased by 92.8 to 100% (mean 96.9 +/- 3.7% [+/- standard error of the mean]) relative to the blood flow before the test. This study demonstrated that the blood flow in the nerve root is reduced when the nerve root is compressed in vivo. CONCLUSIONS: The intraoperative FNST showed that the hernia compressd the nerve roots and there was marked disturbance of gliding, which was reduced to only a few millimeters. During the test, IRBF decreased by 92.8 to 100% (96.9 +/- 3.7% [mean +/- standard error of the mean]). PMID- 14563147 TI - Inhibition of proteoglycan and type II collagen synthesis of disc nucleus cells by nicotine. AB - OBJECT: Systemic nicotine has been hypothesized to cause degeneration of the intervertebral disc which in turn decreases vascular supply to the disc through a cholinergic receptor-mediated process. Another possible mechanism may be through direct regulatory effects on disc cells. In this study, the authors tested the hypothesis that nicotine adversely affects nucleus pulposus cells by directly inhibiting proteoglycan synthesis and gene expression of type II collagen (Phase I study). They also assessed the hypothesis that nicotine inhibits the bone morphogenetic protein (BMP)-2-induced upregulation of extracellular matrix (Phase II study). METHODS: Cells were isolated from nucleus pulposus obtained in rat lumbar discs and cultured on a monolayer. Media were treated with nicotine and/or recombinant human (rh)BMP-2 for 7 days. Sulfated glycosaminoglycan (SO4-GAG) in media was quantified using 1,9-dimethylmethylene blue (DMMB) assay. Gene assay of types I and II collagen, Sox9, and glyceraldehyde-3-phosphate dehydrogenase were quantified using reverse transcriptase-polymerase chain reaction (RT-PCR) and real time PCR. In the Phase I study, nicotine-treated (100 microg/ml) and non treated cells were compared. The s-GAG production and messenger RNA (mRNA) of type II collagen and Sox9 decreased significantly in the nicotine-treated group. In the Phase II study, five groups were compared: 1) non-treatment; 2) rhBMP-2 only (100 ng/ml); and 3-5) with rhBMP-2 (100 ng/ml) and increasing doses of nicotine (1 [third group], 10, [fourth group], 100 [fifth group] microg/ml). The SO4-GAG production and mRNA of type II collagen and Sox9 decreased significantly in the groups treated with rhBMP-2 combined with 10 and 100 microg/ml of nicotine compared with the group treated with rhBMP-2. CONCLUSIONS: The results of this study raise the possibility that nicotine may contribute to the process of disc degeneration by a direct effect on the nucleus pulposus cells, possibly by antagonizing the effect of BMP-2. PMID- 14563149 TI - Upper thoracic spinal cord herniation after traumatic nerve root avulsion. Case report and review of the literature. AB - Transdural herniation of the spinal cord, a rare but well-documented entity, has been reported sporadically for more than 25 years as a possible cause for various neurological signs and symptoms ranging from isolated sensory or motor findings to myelopathy and Brown-Sequard syndrome. The authors report, to the best of their knowledge, the first case of upper thoracic spinal cord herniation occurring after traumatic nerve root avulsion. PMID- 14563150 TI - Fracture of S1-2 after L4-S1 decompression and fusion. Case report and review of the literature. AB - The author describes a woman in whom an S1-2 fracture developed after L4-S1 decompression and fusion. Osteoporosis was not present, but the lesion failed to respond to conservative therapy, necessitating surgical extension of the spinal fusion. Although biomechanical complications of lumbosacral fusion are uncommon, pseudarthrosis, degenerative spondylolisthesis, and stress fractures have been reported. To date, only four cases of sacral stress fracture appear in the literature, all involving female patients and reportedly associated with osteoporosis. Unlike the present case, the fracture resolved satisfactorily in all cases with conservative treatment. PMID- 14563151 TI - Magnetic resonance imaging-guided percutaneous fenestration of a cervical intradural cyst. Case report. AB - The authors describe the case of a high cervical, intradural extramedullary cyst located anterior to the spinal cord in a 13-year-old boy. The lesion was fenestrated percutaneously by using real-time magnetic resonance (MR) imaging guidance and a local anesthetic agent. The patient's symptom, severe exercise induced headache, immediately resolved after treatment. Nine months later, complete disappearance of the cyst was confirmed on MR imaging and computerized tomography myelography. Magnetic resonance imageing-guided fenestration can be considered a minimally invasive option for intradural cystic lesions. PMID- 14563152 TI - Spinal intradural extramedullary cavernous angioma. Case report. AB - The spinal intradural extramedullary cavernous angioma is a rare clinical entity. Only 20 surgically treated cases have been reported. The authors report on an additional case in which the lesion was located in the cervical region, and they summarize its unique clinical features. Intradural extramedullary cavernous angiomas occur predominantly in males, in the lower thoracolumbar region, exhibit a relatively high association with subarachnoid hemorrhage, and mostly adhere to the nerve root or spinal cord. Because resection is possible without causing morbidity and because outcome depends on the severity of preoperative neurological dysfunction, precise diagnosis and timely treatment are mandatory. PMID- 14563153 TI - Posterolateral percutaneous endoscopic lumbar foraminotomy for L5-S1 foraminal or lateral exit zone stenosis. Technical note. AB - The purpose of this study was to determine the efficacy and feasibility of posterolateral percutaneous endoscopic lumbar foraminotomy (PELF) for foraminal or lateral exit zone stenosis of the L5-S1 level in the awake patient. Twelve consecutive patients with L5-S1 foraminal stenosis and associated leg pain underwent PELF between May 2001 and July 2002. Under fluoroscopic guidance, posterolateral endoscopic foraminal decompression was performed using a bone reamer, endoscopic forceps, and a laser. Using this new technique, the authors removed part of the hypertrophied superior facet, thickened ligamentum flavum, and protruded disc compressing the exiting (L-5) nerve root. Clinical outcome was measured using the Macnab criteria. The mean follow-up period was 12.9 months. All the patients were discharged within 24 hours. Satisfactory (excellent or good) results were demonstrated in 10 patients. There was no complication. The PELF procedure provides a simple alternative for treating lumbar foraminal or lateral exit zone stenosis in selected cases. The authors found that the posterolateral endoscopic approach to the L5-S1 foramen was usually possible and that using a bone reamer to undercut the superior facet was effective. PMID- 14563154 TI - Three-dimensional fluoroscopy-guided percutaneous thoracolumbar pedicle screw placement. Technical note. AB - The authors sought to evaluate the feasibility and accuracy of three-dimensional (3D) fluoroscopic guidance for percutaneous placement of thoracic and lumbar pedicle screws in three cadaveric specimens. After attaching a percutaneous dynamic reference array to the surgical anatomy, an isocentric C-arm fluoroscope was used to obtain images of the region of interest. Light-emitting diodes attached to the C-arm unit were tracked using an electrooptical camera. The image data set was transferred to the image-guided workstation, which performed an automated registration. Using the workstation display, pedicle screw trajectories were planned. An image-guided drill guide was passed through a stab incision, and this was followed by sequential image-guided pedicle drilling, tapping, and screw placement. Pedicle screws of various diameters (range 4-6.5 mm) were placed in all pedicles greater than 4 mm in diameter. Postoperatively, thin-cut computerized tomography scans were obtained to determine the accuracy of screw placement. Eighty-nine (94.7%) of 94 percutaneous screws were placed completely within the cortical pedicle margins, including all 30 lumbar screws (100%) and 59 (92%) of 64 thoracic screws. The mean diameter of all thoracic pedicles was 6 mm (range 2.9-11 mm); the mean diameter of the five pedicles in which wall violations occurred was 4.6 mm (range 4.1-6.3 mm). Two of the violations were less than 2 mm beyond the cortex; the others were between 2 and 3 mm. Coupled with an image guidance system, 3D fluoroscopy allows highly accurate spinal navigation. Results of this study suggest that this technology will facilitate the application of minimally invasive techniques to the field of spine surgery. PMID- 14563156 TI - The novel gluconeogenesis inhibitors FR225659 and related compounds that originate from Helicomyces sp. No. 19353. I. Taxonomy, fermentation, isolation and physico-chemical properties. AB - FR225659 and four related compounds are novel gluconeogenesis inhibitors that consist of a novel acyl-group and three abnormal amino acids. They were isolated from the culture broth of Helicomyces sp. No. 19353 and can be purified by absorptive resin and reverse-phase column chromatography. They are potent inhibitors of gluconeogenesis in primary cultured rat hepatocytes and thus may be useful as anti-diabetic agents. PMID- 14563155 TI - New cell-cell adhesion inhibitors from Streptomyces sp. UMA-044. AB - Three compounds, NP25301 (1), NP25302 (2) and bohemamine (3), inhibitors of cell adhesion based on LFA-1/ICAM-1, were isolated from the cultured broth of the strain Streptomyces sp. UMA-044. New compounds 1 and 2 were identified as 2-(3' carbamoylphenoxy)acrylic acid methyl ester and deoxybohemamine, respectively, based on spectroscopic analyses. Compounds 1 approximately 3 inhibited adhesion of HL-60 cells to CHO-ICAM-1 cells at IC50 values of 29.5 microg/ml for 1, 24.3 microg/ml for 2, and 27.2 microg/ml for 3. PMID- 14563157 TI - The novel gluconeogenesis inhibitors FR225659 and related compounds that originate from Helicomyces sp. No. 19353. II. Biological profiles. AB - The novel gluconeogenesis inhibitor FR225659 and four related compounds were isolated from the cultured broth of a fungal strain No. 19353. These compounds inhibited the glucagon-stimulated gluconeogenesis of rat primary hepatocytes and had hypoglycemic effects in two different in vivo models. PMID- 14563158 TI - The novel gluconeogenesis inhibitors FR225659 and FR225656 from Helicomyces sp. No. 19353. III. Structure determination. AB - During the course of screening for novel gluconeogenesis inhibitors, FR225659 and its related compounds were isolated from a fermentation broth of Helicomyces sp. No. 19353. Spectroscopic analysis concluded that FR225659 is an N-acyl tripeptide consisting of a novel acyl, a 3-chloro-4-hydroxyarginine, a 3-hydroxy-3 methylproline and a dehydrovaline. Degradation study allowed assignment of the absolute configuration of the 3-hydroxy-3-methylproline to be (2S,3R). FR225656 was shown to possess a dehydroisoleucine instead of the dehydrovaline of FR225659. PMID- 14563159 TI - Clethramycin, a new inhibitor of pollen tube growth with antifungal activity from Streptomyces hygroscopicus TP-A0623. I. Screening, taxonomy, fermentation, isolation and biological properties. AB - The cytoskeletal proteins, actin and myosin, play a central role in pollen tube growth. The pollen tube growth is inhibited by cytochalasin, which interferes with actin polymerization. In the screening of pollen tube growth inhibitors, clethramycin was found from the fermentation broth of an actinomycete strain TP A0623. The producing strain was isolated from a root of Clethra barbinervis collected in Toyama, Japan and identified as Streptomyces hygroscopicus based on the taxonomic study. Clethramycin showed in vitro antifungal activity against yeast such as Candida albicans and C. glabrata with the MIC of 0.5 approximately 8 microg/ml, but weak activity against Gram-positive and negative bacteria (MIC > or = 64 microg/ml). Cytotoxicity of clethramycin was moderate and the IC50 was 57 microg/ml against HeLa cells and 120 microg/ml against WI-38 cells. PMID- 14563160 TI - Clethramycin, a new inhibitor of pollen tube growth with antifungal activity from Streptomyces hygroscopicus TP-A0623. II. Physico-chemical properties and structure determination. AB - Clethramycin is a novel linear polyene polyketide produced by a plant-associated actinomycete Streptomyces hygroscopicus TP-A0623. It possesses a guanidino and carboxyl residue at each end and an O-sulfate group and a hexaene moiety in the molecule, and its molecular formula is C63H99N3O18S. The structure was determined by analyzing 2D-NMR and FAB-MS data, using 13C-enriched compounds. Clethramycin is structurally similar to linearmycin, an inhibitor of spheroplast regeneration in Candida albicans. Clethramycin shows potent antifungal and pollen tube growth inhibitory activity. PMID- 14563161 TI - Anti-angiogenesis effects of borrelidin are mediated through distinct pathways: threonyl-tRNA synthetase and caspases are independently involved in suppression of proliferation and induction of apoptosis in endothelial cells. AB - Borrelidin, an antibiotic with anti-angiogenic activity, not only suppresses new capillary tube formation, but also collapses formed capillary tubes in a rat aorta culture model. Since it selectively inhibits threonyl-tRNA synthetase, we examined the effect of threonine on its anti-angiogenic activity. We found that a high concentration of threonine (1 mM) attenuated the ability of borrelidin to inhibit both capillary tube formation in the rat aorta culture model and human umbilical vein endothelial cells (HUVEC) proliferation, yet did not affect the ability of borrelidin to collapse formed capillary tubes or to induce apoptosis in HUVEC. Borrelidin activated caspase-3 and -8, and inhibitors of both caspase-3 and -8 suppressed borrelidin-induced apoptosis in HUVEC. Taken together, these data suggest that the anti-angiogenic effects of borrelidin are mediated through at least two mechanisms, i.e. one threonine-dependent and the other threonine independent, and borrelidin induces apoptosis in endothelial cells via the caspase-8/-3 pathway. PMID- 14563163 TI - Effect of 14-membered macrolide compounds on monocyte to macrophage differentiation. PMID- 14563162 TI - SNA-60-367 components, new peptide enzyme inhibitors of aromatase: structure of the fatty acid side chain and amino acid sequence by mass spectrometry. AB - SNA-60-367 components, new peptide enzyme inhibitors of aromatase, were isolated from the culture broth of soil bacterium, Bacillus sp. SNA-60-367. These inhibitors are a family of acylated decapeptides that differ from each other in terms of amino acid composition and the nature of the fatty acid side chain. The structures of the fatty acid moieties were shown to be (3-hydroxy)heptadecanoic acid and (3-hydroxy)hexadecanoic acid that possess normal-, iso- or anteiso-type alkyl groups. The amino acid sequence of the open form of the lactone ring of the acylpeptides is RCO-L-Glu-D-Orn-L(or D)-Tyr3-D-allo-Thr-L-Glu-D-X1 (Ala, Aba or Val)-L-Pro-L-Gln-D(or L)-Tyr-L-X2(10)(Ile or Val)-OH. The lactone ring of SNA-60 367 components is formed between Tyr3 and X2(10). PMID- 14563164 TI - Inhibition of CTP: phosphocholine cytidylyltransferase activity by 15 deoxyspergualin. PMID- 14563165 TI - In vitro and in vivo antimalarial activities of a non-glycosidic 18-membered macrolide antibiotic, borrelidin, against drug-resistant strains of Plasmodia. PMID- 14563166 TI - Total syntheses of the AChE inhibitors (-)-arisugacins F and G. PMID- 14563167 TI - The accidental protozoologist--my journey through the world of ciliates. AB - The Past-President's Address has been the opportunity for the speaker to reminisce about the road traveled to get to this time in life. In this paper, I continue in that tradition. During my journey to the present day, I visited different laboratories, studying the genetics of mating and mating types in Paramecium, Tetrahymena, Blepharisma and Euplotes. I have met and worked with many distinguished scientists, including other Past-Presidents of the Society. I also became an active participant in the Society of Protozoologists. I hope the recounting of my trip will be both entertaining and enlightening. PMID- 14563168 TI - Lack of arginine decarboxylase in Trypanosoma cruzi epimastigotes. AB - The presence of arginine decarboxylase (ADC) enzymatic activity in Trypanosoma cruzi epimastigotes is still a matter of controversy due to conflicting results published during the last few years. We have investigated whether arginine might indeed be a precursor of putrescine via agmatine in these parasites. We have shown that wild-type T. cruzi epimastigotes cultivated in a medium almost free of polyamines stopped their growth after several repeated passages of cultures in the same medium, and that neither arginine nor omithine were able to support or reinitiate parasite multiplication. In contrast, normal growth was quickly resumed after adding exogenous putrescine or spermidine. The in vivo labelling of parasites with radioactive arginine showed no conversion of this amino acid into agmatine, and attempts to detect ADC activity measured by the release of CO2 under different conditions in T. cruzi extracts gave negligible values for all strains assayed. The described data clearly indicate that wild-type T. cruzi epimastigotes lack ADC enzymatic activity. PMID- 14563169 TI - Isolation and characterization of glutathione reductase from Physarum polycephalum and stage-specific expression of the enzyme in life-cycle stages with different oxidation-reduction levels. AB - Physarum polycephalum has a life cycle with several distinct phases that have different oxidation-reduction requirements. To investigate the relationship between the life cycle and the oxidation-reduction state, we isolated glutathione reductase (GR; EC 1.6.4.2) from Physarum microplasmodia. The enzyme was found to be a homodimer with a subunit M(r) of 49,000, and K(m) values for oxidized glutathione and NADPH of 40 and 28.6 microM, respectively. We then constructed a cDNA library from microplasmodium mRNA and cloned GR cDNA from the library. The isolated cDNA consisted of 1,475 bp encoding a polypeptide of 452 amino acids. The amino acid sequence similarity was about 50% with GRs of other organisms, and several conserved sequence motifs thought to be necessary for activity are evident in the Physarum enzyme. Escherichia coli transformed with an expression vector containing the cDNA synthesized the active GR. Genomic Southern blot analysis indicated that the GR gene is present as a single copy in the Physarum genome. Immunoblot analysis and RT-PCR analysis detected GR mRNA expression in the microplasmodium, plasmodium, and sclerotium, but not in the spore or flagellate. GR activity was low in the spore and flagellate. These results suggest that the glutathione oxidation-reduction system relates to the Physarum life cycle. PMID- 14563170 TI - Ultrastructural responses in field-bleached and experimentally stressed Amphistegina gibbosa (Class Foraminifera). AB - Amphistegina are the most common foraminifers with algal endosymbionts found on reefs and carbonate shelves worldwide. Like zooxanthellate corals and other reef organisms with algal symbionts, Amphistegina respond to photoxidative stress by bleaching. This paper documents ultrastructural changes that occur during bleaching under field and laboratory conditions. Nine chambers from the outer whorl of each of 22 normal-appearing and 11 partly bleached specimens of Amphistegina gibbosa, which were collected from Conch Reef, Florida, USA, were examined using transmission electron microscopy. The condition and numbers of algal symbionts, as well as the cell area occupied by 10 other intracellular structures of the host, were quantified. Normal-appearing specimens averaged three times more viable symbionts and less than a fourth as many deteriorating symbionts as partly bleached specimens. Foraminifers experimentally exposed to visible light intensities > or = 13 micromole photon m(-2) s(-1) for 35 d were statistically similar to partly bleached field specimens in the number and condition of symbionts, and in chamber area occupied by the evaluated host structures. Exposure to 32 degrees C water temperature at 6-8 micromole photon m( 2) s(-1) for 28 d induced symbiont loss but did not degrade host endoplasm. PMID- 14563171 TI - Molecular phylogeny and surface morphology of Colpodella edax (Alveolata): insights into the phagotrophic ancestry of apicomplexans. AB - The molecular phylogeny of colpodellids provides a framework for inferences about the earliest stages in apicomplexan evolution and the characteristics of the last common ancestor of apicomplexans and dinoflagellates. We extended this research by presenting phylogenetic analyses of small subunit rRNA gene sequences from Colpodella edax and three unidentified eukaryotes published from molecular phylogenetic surveys of anoxic environments. Phylogenetic analyses consistently showed C. edax and the environmental sequences nested within a colpodellid clade, which formed the sister group to (eu)apicomplexans. We also presented surface details of C. edax using scanning electron microscopy in order to supplement previous ultrastructural investigations of this species using transmission electron microscopy and to provide morphological context for interpreting environmental sequences. The microscopical data confirmed a sparse distribution of micropores, an amphiesma consisting of small polygonal alveoli, flagellar hairs on the anterior flagellum, and a rostrum molded by the underlying (open sided) conoid. Three flagella were present in some individuals, a peculiar feature also found in the microgametes of some apicomplexans. PMID- 14563172 TI - Protein binding to expanded telomere repeats in Tetrahymena thermophila. AB - The ends of eukaryotic chromosomes are protected by DNA-protein structures called telomeres. Telomeric DNA is highly conserved, usually consisting of long tracts of a repeating G-rich sequence. Tetrahymena thermophila telomeric DNA consists of alternating blocks of GGGG and TT sequences (i.e. a G4T2 repeat sequence). We examined the relative importance of the guanine and thymine elements of the repeat sequence in promoting in vitro binding by T. thermophila proteins. We identified single- and, for the first time, double-stranded telomere binding activities from a crude T. thermophila protein extract and tested the binding of these activities to altered telomere repeat sequences. All deletions or substitutions made to the guanine element virtually abolished binding, indicating that four G's are essential for recognition by the binding activity. However, G's alone are not sufficient for efficient binding, as elimination of the thymine element dramatically reduced binding. By contrast, substantial expansion of the thymine element was well tolerated, even though one such change, G4T4, is lethal in vivo. We tested up to a four-fold expansion of the thymine element and found that highly efficient binding was still achieved. These results suggest a minimal recognition sequence for T. thermophila proteins, with the T element providing an important spacer between essential G elements. PMID- 14563174 TI - Interstitial ciliates: benthic microaerophiles or planktonic anaerobes? AB - We observed marine benthic interstitial ciliates Geleia sp. and Tracheloraphis sp. inhabiting the water column of a chemically stratified salt pond. This habitat is uncharacteristic for interstitial ciliates, yet they displayed active and abundant planktonic populations (up to 800 and 250 cells/liter, respectively) and a well-defined pattern of vertical distribution. Completely absent from the oxygenated epilimnion, they first appeared at the oxic/anoxic interface and were present throughout the anoxic hypolimnion. The data could not be explained by a passive removal (e.g. by currents) of these ciliates from their conventional habitat (soft sediments) to water column. The results suggest that 1) these ciliates favored an anoxic environment, and 2) they switched to a planktonic lifestyle as appropriate conditions (seasonal anoxia) developed in the water column. This sharply contrasts the classic view of these ciliates as specifically benthic and aerobic (albeit microaerophilic) organisms. We hypothesize that Geleia sp. and Tracheloraphis sp. can readily grow in either water column or benthos, but are typically found in sediments simply because they contain their preferred (anoxic) niche. PMID- 14563173 TI - G-protein modulators alter the swimming behavior and calcium influx of Paramecium tetraurelia. AB - To assess the potential role of G-proteins in chemokinesis, Paramecium tetraurelia was pre-incubated with the G-protein modulator pertussis toxin. Pertussis toxin pretreatment significantly reduced Paramecium chemoattraction to sodium acetate and ammonium chloride in T-maze behavioral assays and depressed the frequency of avoidance reactions, indicating that heterotrimeric G-proteins may be involved with the motility response. To determine whether G-proteins exert their effect via the ciliary voltage-sensitive calcium channel, we examined responses of P. tetraurelia to the potent voltage-sensitive calcium channel agonist, deltamethrin. Pertussis toxin preincubation significantly reduced the toxic effects of deltamethrin exposure as determined by survival under depolarizing conditions and reduced the duration of backward swimming episodes in behavioral bioassays. Furthermore, non-hydrolyzable analogs of guanine nucleotides altered deltamethrin-stimulated calcium influx via calcium channels in isolated ciliary vesicles. Heterotrimeric G-protein subunits were subsequently detected in ciliary vesicles of P. tetraurelia by antibodies produced against Galpha and Gbeta subunits, and by 32P-ADP-ribosylation, indicating that proteins of the appropriate molecular weight are the target of pertussis toxin in these vesicles. These findings provide additional evidence that heterotrimeric G proteins are associated with ciliary vesicles and that they play a role in the modulation of swimming behavior and the toxic action of deltamethrin in Paramecium. PMID- 14563175 TI - Description of a marine peritrichous ciliate, Pseudovorticella sinensis n. sp. (Ciliophora, Peritrichia) from China. AB - A new marine peritrich ciliate, Pseudovorticella sinensis n. sp. was isolated from a shrimp-farming pond in the littoral area of Qingdao, China. The morphology, infraciliature, and silverline system were studied based on living and silver-impregnated specimens. This species is characterized by (1) an elongated bell-shaped body that measures 50-60 x 35-45 microm in vivo, (2) one large, ventrally located contractile vacuole, and (3) a pellicle covered by a layer of transparent, cortical vesicles. The number of transverse silverlines from the peristomial area to the aboral ciliary wreath is 26-32, and from the aboral ciliary wreath to the scopula is 12-15. The stalk measures about 160-250 microm long x 5-6 microm wide. The spasmoneme has one row of conspicuous thecoplasmic granules, which are about 0.8 microm in diameter. PMID- 14563176 TI - Glycogen phosphorylase sequences from the amitochondriate protists, Trichomonas vaginalis, Mastigamoeba balamuthi, Entamoeba histolytica and Giardia intestinalis. AB - Glycogen phosphorylase genes or messages from four amitochondriate eukaryotes, Trichomonas vaginalis, Mastigamoeba balamuthi, Entamoeba histolytica (two genes) and Giardia intestinalis, have been isolated and sequenced. The sequences of the amitochondriate protist enzymes appear to share a most recent common ancestor. The clade containing these sequences is closest to that of another protist, the slime mold (Dictyostelium discoideum), and is more closely related to fungal and plant phosphorylases than to mammalian and eubacterial homologs. Structure-based amino acid alignment shows conservation of the residues and domains involved in catalysis and allosteric regulation by glucose 6-phosphate but high divergence at domains involved in phosphorylation-dependent regulation and AMP binding in fungi and animals. Protist phosphorylases, as their prokaryotic and plant counterparts, are probably not regulated by phosphorylation. PMID- 14563177 TI - Isolation of a thermotolerant Paravahlkampfia sp. from lizard intestine: biology and molecular identification. AB - An amoeba was isolated from the intestines of several moribund pink-tongued skinks (lizards), Hemisphaeriodon gerrardi. Unusual features of this isolate were its ability to grow at temperatures of > or = 37 degrees C, and its inability to use Escherichia coli as a food source or to grow axenically on a variety of enriched culture media suitable for other soil amoeba isolates. Growth was abundant, however, on tissue culture cells, with amoebae clearing cell monolayers in approximately 48 h at 37 degrees C. Trophozoites had a vahlkampfiid-like morphology, moving by means of an anterior eruptive pseudopod. Cysts, round to slightly ovoid and lacking exit pores, were formed in culture. Tests for enflagellation of trophic amoebae were negative. Indirect immunofluorescence staining was negative for Naegleria fowleri and Willaertia sp. The isolate was sensitive to azithromycin, but not to amphotericin B, pentamidine isethionate, fluconazole, 5-fluorocytosine, and sulfadiazine. Phylogenetic analysis based on the PCR-amplified small subunit ribosomal DNA, identified the organism as Paravahlkampfia ustiana, an amoeba not previously isolated from either poikilothermic or homeothermic hosts. No evidence of pathology was seen in stained sections of lizard intestine, suggesting that the ameba was part of the normal fauna of the lizard gut. Its diet in the lizard intestine is unknown and the organism may have unusual growth requirements. Thus, P. ustiana joins other soil amoebae that have been isolated from mammals, amphibia, fish, and reptiles, which have the potential of becoming opportunistic pathogens. PMID- 14563178 TI - Role of gregarine parasite Ascogregarina culicis (Apicomplexa: Lecudinidae) in the maintenance of Chikungunya virus in vector mosquito. AB - Ascogregarina culicis and Ascogregarina taiwanensis are common gregarine parasites of Aedes aegypti and Aedes albopictus mosquitoes, respectively. These mosquito species are also known to transmit dengue and Chikungunya viruses. The sporozoites of these parasites invade the midgut epithelial cells and develop intracellularly and extracellularly in the gut to complete their life cycles. The midgut is also the primary site for virus replication in the vector mosquitoes. Therefore, studies were carried out with a view to determine the possible role of these gregarines in the vertical transmission of dengue and Chikungunya viruses from larval to adult stage. Experiments were performed by exposing first instar mosquito larvae to suspensions containing parasite oocysts and viruses. Since Ascogregarina sporozoites invade the midgut of first instar larvae, the vertical transmission was determined by feeding the uninfected first instar larvae on the freshly prepared homogenates from mosquitoes, which were dually infected with viruses and the parasite oocysts. Similarly, the role of protozoan parasites in the vertical transmission of viruses was determined by exposing fresh first instar larvae to the dried pellets of homogenates prepared from the mosquitoes dually infected with viruses and the parasite oocysts. Direct vertical transmission and the vertical transmission of CHIK virus through the oocyst of the parasites were observed in the case of Ae. aegypti mosquitoes. It is suggested that As. culicis may have an important role in the maintenance of CHIK virus during the inter-epidemic period. PMID- 14563179 TI - Weight-based dosing of pegylated interferon-alpha in chronic hepatitis C: just a marketing 'gag'? AB - Today medical-scientific data are diluted by the marketing strategies of the biomedical industry making it difficult for practising physicians to decide what is correct or wrong. One typical example is the use of pegylated interferons for treatment of chronic hepatititis C. In this report the arguments pro and contra weight-based dosing are critically discussed. The factors contributing to success or failure to eradicate the virus are manifold, and include the sensitivity of the virus to interferon, viral genotype, age, gender stage of fibrosis, presence or absense of steatosis. Weight by itself plays just a minor role. The impact of weight-based dosing in general is overestimated and certainly not needed when 40 kD branched PEG-IFNalpha2a with a restricted volume of distribution is used. Whether weight-based dosing of 12 kD linear PEG-IFNalpha2b provides any benefit over a flat dose of the drug remains to be studied. PMID- 14563180 TI - Determination of faecal calprotectin, a novel marker of organic gastrointestinal disorders. PMID- 14563181 TI - Is there any remaining role for methotrexate for Crohn's disease? PMID- 14563182 TI - Breath test is very reliable for diagnosis of Helicobacter pylori infection in real clinical practice. AB - AIMS: To determine the accuracy of the most common available tests for the diagnosis of Helicobacter pylori infection in an unselected and untreated population of patients. PATIENTS AND METHODS: Prospective study including 314 unselected patients from a population of 814 patients referred for upper endoscopy at one hospital. H. pylori infection was diagnosed by rapid urease test (RUT), histology, culture and 13C-urea-breath test (UBT) and serum IgG (EIA). H. pylori infection was defined as positive if culture or at least two of the other tests were positive. RESULTS: The prevalence of H. pylori infection in this population was 72%. The diagnostic test with the greatest combination of sensitivity (97%) and specificity (100%) was the UBT. EIA had a good sensitivity (96%), but it was the test with the least specificity (71%). RUT, histology and culture showed a high specificity (>98%) but a sensitivity lower than 90%. In elderly patients (>65 years old, n=120), UBT was also the test with the greatest combination of sensitivity (94%) and specificity (100%). CONCLUSIONS: In conditions of real clinical practice the 13C-urea-breath test is a reliable test for H. pylori diagnosis, both in young and elderly patients. PMID- 14563183 TI - Comparison between methotrexate and azathioprine in the treatment of chronic active Crohn's disease: a randomised, investigator-blind study. AB - BACKGROUND AND AIMS: The efficacy of azathioprine in the treatment of chronic active Crohn's disease is well established. However, this drug has a long onset of action. Methotrexate has also been shown to be effective in chronic active Crohn's disease. The aim of this study was to evaluate the efficacy and safety of methotrexate in comparison with azathioprine, and to establish whether methotrexate has a shorter onset of action in this setting. METHODS: Patients with chronic active Crohn's disease were admitted to this investigator-blind study. Chronicity was defined as the need for steroid therapy of > or = 10 mg/day for at least 4 months during the preceding 12 months, with at least one attempt to discontinue treatment. The disease had to be clinically active at entry, with a Crohn's Disease Activity Index of > or = 200. Six patients treated with azathioprine and methotrexate, respectively, were found to have enterocutaneous and perianal fistulas. At entry, all patients received prednisolone (40 mg once a day) which was tapered over a period of 12 weeks unless their clinical condition deteriorated. All patients were randomised to receive i.v. methotrexate 25 mg/week, or oral azathioprine 2 mg/kg per day, for a 6-month follow-up period. After the first 3 months, methotrexate was switched to oral administration maintaining the same dose. The primary efficacy outcome considered was the proportion of patients entering first remission after 3 and 6 months of therapy. Clinical remission was defined as the lack of need for steroid treatment and a Crohn's Disease Activity Index score of < or = 150 points at each scheduled visit. RESULTS: In the 54 patients (26 F, 28 M, mean age 34 years, range 18-60) randomly assigned to methotrexate (n=27) or azathioprine (n=27), no statistically significant difference was found between the two treatment regimens with respect to remission rate after 3 (methotrexate 44%, azathioprine 33%, p=0.28, (95% CI, 0.369-0.147), and 6 months (methotrexate 56%, azathioprine 63%, p=0.39, 95% CI, 0.187-0.335), respectively. Six patients withdrew from therapy due to adverse events: 3/27 (11%) in methotrexate and 3/27 (11%) in azathioprine. Drug-related adverse events (asthenia, nausea and vomiting) that did not require withdrawal from therapy were more frequent in the methotrexate group (azathioprine: 2/27 (7%); methotrexate: 12/27 (44%), p=0.00009). The frequency of these adverse events was comparable during the intravenous or oral administration of the drug. CONCLUSIONS: This study confirms that methotrexate is effective in inducing remission in patients with chronic active Crohn's disease, therapeutic efficacy being comparable, but not faster, than that of azathioprine. PMID- 14563184 TI - Colorectal cancer and high grade dysplasia complicating ulcerative colitis in Italy. A retrospective co-operative IG-IBD study. AB - BACKGROUND: Ulcerative colitis is a well-known risk factor for colorectal cancer. AIM: To take a census of the cases of colorectal cancer in ulcerative colitis patients observed in Italy and to evaluate the clinical presentation of neoplastic complication. PATIENTS AND METHODS: Experts from 28 Italian centres specialised in the management of inflammatory bowel disease or malignancies participated to the study. They were invited to send clinical data of patients with ulcerative colitis complicated by colorectal cancer or high-grade dysplasia consecutively observed between 1985 and 2000. One hundred and twelve patients (92 with cancer and 20 with high-grade dysplasia) were collected. Fourteen of them had undergone colectomy and ileo-rectal anastomosis for ulcerative colitis. Data of surgical patients were analysed separately. RESULTS: The mean age at diagnosis of ulcerative colitis and colorectal cancer patients was 39.3 and 53.2 years, respectively, and the mean duration between diagnosis of ulcerative colitis and cancer was 13.9 years (range 0-53). Inflammation was proximal to the splenic flexure in 71 cases (76.3%). One hundred and three colorectal cancers were registered (93 patients with single lesion and five patients with two synchronous cancers), with 76.7% of cancers being located in the left colon. As to the surgical patients, the mean age at diagnosis of ulcerative colitis and cancer was 28.9 and 47.0 years, respectively, and the mean diagnostic interval for ulcerative colitis and cancer was 18.1 years. Only 51 out of 112 patients were in follow-up. An early diagnosis of neoplasia (high grade dysplasia, stage A or B sec. Dukes) occurred in 72.5% of patients who were subjected to endoscopic surveillance and in 48.0% of patients who did not undergo endoscopic surveillance (p=0.02). CONCLUSIONS: These data show an earlier diagnosis of cancer in patients who had undergone endoscopic surveillance. The poor compliance to the follow-up program, however, reduces its effectiveness. Moreover, total colectomy allows an easier follow-up, with only the rectum being controlled. Colectomy with ileo rectal anastomosis or proctocolectomy with ileo-anal anastomosis, could represent a valid alternative in patients at high risk of cancer who refuse endoscopic surveillance. PMID- 14563185 TI - Glutathione supplementation improves oxidative damage in experimental colitis. AB - BACKGROUND: The pathogenesis of inflammatory bowel disease is due, in part, to enhanced free-radical production and reduced antioxidant potential in mucosa cells. AIM: We evaluated in a rat model of trinitrobenzensulphonic acid (TNBS) colitis to see whether parenteral administration of glutathione is able to improve mucosal oxidative damage at onset (study A) and during chronic phases of colitis (study B). METHODS: In study A, the rats were injected with a single dose of glutathione (200 mg/kg, i.p.) or saline (0,2 ml, i.p.) 1 h before colitis induction and killed 1 h later. In study B, rats with induced colitis were treated with daily injection of glutathione (50 mg/kg, i.p.) or saline (0,2 ml, i.p.), and killed at 1, 2, 4 and 8 weeks. We evaluated on mucosal samples the macroscopic and histological damage and the oxidative stress assessed by the mucosal levels of lipoperoxides, malonyldialdehyde, glutathione and cysteine. RESULTS: In study A, colitis induction caused a significant increase to the total histological score (p<0.05), lipoperoxide and malonyldialdehyde levels (p<0.001), but did not affect glutathione and cysteine content. Glutathione pre-treatment decreased both total histological score (p<0.05) and lipoperoxide and malonyldialdehyde values (p<0.001). In study B, the extensive macroscopic and histological colonic damage induced by TNBS was accompanied by a reduction of glutathione and cysteine mucosal levels (p<0.01) and increased lipid peroxidation. Glutathione supplementation significantly improved colonic damage (p<0.01), restored glutathione and cysteine levels, and decreased, and even, if not totally, abolished lipid peroxidation (p<0.001). CONCLUSION: This paper further supports the pathogenic role of the imbalance in oxidant/antioxidant content in inducing mucosal colonic damage. PMID- 14563186 TI - Role of faecal calprotectin as non-invasive marker of intestinal inflammation. AB - BACKGROUND/AIM: Faecal calprotectin, a neutrophil granulocyte cytosol protein, is considered a promising marker of intestinal inflammation. We assessed and compared the faecal calprotectin concentration in patients with organic and functional chronic intestinal disorders. PATIENTS AND METHODS: The study was carried out, using a commercially available ELISA test, measuring calprotectin in stool samples collected from 131 patients with inflammatory bowel diseases, 26 with intestinal neoplasms, 48 with irritable bowel syndrome and 34 healthy subjects. RESULTS: Median faecal calprotectin was significantly increased in Crohn's disease (231 microg/g, 95% confidence interval (CI) 110-353 microg/g), ulcerative colitis (167 microg/g, 95% CI 59-276 microg/g), and neoplasms (105 microg/g, 95% CI 0-272 microg/g), whereas normal values were found in patients with irritable bowel syndrome (22 microg/g, 95% CI 9-35 microg/g) and in healthy subjects (11 microg/g, 95% CI 3-18 microg/g). A positive correlation was observed with clinical activity scores in Crohn's disease and ulcerative colitis. In both groups, patients with clinically active disease showed higher calprotectin levels than those observed in patients with quiescent disease (405 microg/g, 95% CI 200 610 microg/g vs. 213 microg/g, 95% CI 85-341 microg/g in CD patients, p<0.05, and 327 microg/g, 95% CI 104-550 microg/g vs. 123 microg/g, 95% CI 40-206 microg/g in UC patients, p<0.001). CONCLUSIONS: Faecal calprotectin appears to be a promising and non-invasive biomarker of intestinal inflammation. If these findings are confirmed, it may provide a useful test for the diagnosis and follow up of inflammatory bowel diseases. PMID- 14563187 TI - Body mass index is a good predictor of an elevated alanine transaminase level in the general population: hints from the Dionysos study. AB - AIM: To establish the contribution of body mass index (BMI), sex, age, ethanol intake, hepatitis B (HBV) and hepatitis C (HCV) virus infection, coffee and drug consumption, and cigarette smoking to account for an elevated alanine transaminase (ALT) level in the general population. SUBJECTS: A total of 6315 adult subjects from the Dionysos study. METHODS: Logistic regression was used to quantify the contribution of the variables of interest to elevated ALT, defined as a value of ALT>60 U/l. Areas under ROC curves (AUCs) were calculated to assess accuracy. RESULTS: All the variables considered, with the exception of coffee and drug consumption, were significant predictors of elevated ALT at univariable analyses. When significant predictors were employed in a multivariable model, age and cigarette smoking were no longer significant. The AUC was 0.77 (95% CI=0.74 0.80) for the multivariable model and 0.64 (95% CI=0.60-0.68) for the univariable BMI model (p<0.0001 for the comparison). CONCLUSION: BMI is a good predictor of elevated ALT serum activity in the general population. The ability to predict an elevated ALT is however increased substantially by considering sex, ethanol intake, HBV and HCV infection together with BMI. PMID- 14563188 TI - Relapses of biliary acute pancreatitis in patients with previous attack of biliary pancreatitis and gallbladder in situ. AB - AIM: To evaluate relapse of acute pancreatitis in patients with biliary pancreatitis in whom coexisting diseases or patient refusal have excluded cholecystectomy. PATIENTS AND METHODS: Forty-seven patients presenting a first episode of biliary acute pancreatitis underwent urgent endoscopic retrograde cholangiopancreatography (ERCP) with endoscopic sphincterotomy (ES). RESULTS: ERCP with ES was successful in all but one patient (97.8%) who was then cholecystectomised. Complications related to the endoscopic procedure were reported in five patients (10.6%). During the follow-up period (median time 12 months; range 1-84 months), 10 patients (21%) suffered from biliary complications. Three patients (6.4%), all with lithiasis of the gallbladder, had relapses of acute pancreatitis, two of them within 2 months of the previous episode, and one about a year later after ingestion of a rich meal and alcoholic beverages. The first two were cholecystectomised. Two patients died during the follow-up period from unrelated diseases. CONCLUSIONS: In subjects who are at high risk for anaesthesia, endoscopic procedures may be utilised. PMID- 14563189 TI - Acute biliary pancreatitis in a 9-year-old child treated with endoscopic sphincterotomy. AB - Acute biliary pancreatitis is a well recognized complication of gallstone disease in adults. Acute pancreatitis in childhood is usually caused by congenital anomalies of the pancreatico-biliary ducts, viral infections, drug toxicity or abdominal trauma. We report the case of a 9-year-old girl with acute biliary pancreatitis and cholangitis. On urgent endoscopic retrograde cholangiopancreatography a bulging papilla with impacted stone was seen. She was treated with endoscopic sphincterotomy without complications. The disease resolved rapidly and uneventfully after the endoscopic treatment. PMID- 14563190 TI - Is the use of albumin of value in the treatment of ascites in cirrhosis? The case in favour. AB - In patients with cirrhosis, ascites accumulates because of sodium retention, triggered by a reduction of the effective arterial blood volume, and imbalanced Starling forces in the splanchnic area due to portal hypertension and hypoalbuminemia. Albumin is the ideal plasma expander in this setting, since it ameliorates systemic and reneal haemodynamics, so reducing sodium retention, and increases oncotic pressure in the splanchnic compartment. In particular, albumin proved useful in patients treated with diuretics, as demonstrated by a randomised study performed at our Instituition in which 126 ascitic inpatients were treated according to a stepped-care diuretic regimen. In fact, patients receiving diuretics plus albumin (n = 63) had a higher cummulative rate of response (p < 0.05) and a shorter hospital stay (20 +/- 1 versus 24 +/- 2 days, p < 0.05) than those given diuretics alone. Treatment with albumin on an outpatient basis (25 g/week) resulted in a lower probability of developing ascites (p < 0.02 vs. patients not given albumin) and a lower probability of readmission (p < 0.02). Patients given albumin also had a better quality of life. As discussed in another article, evidence also supports the use of albumin in patients treated for paracentesis, as well as in patients with spontaneous peritonitis or hepatorenal syndrome. PMID- 14563191 TI - Is the use of albumin of value in cirrhosis? The case not so in favour, or is there an alternative? AB - A decreased effective arterial blood volume is the principal haemodynamic disturbance in cirrhosis, leading to activation of the renin angiotensin aldosterone and the sympathetic nervous systems, sodium and water retention and renal impairment. Albumin is a plasma expander that could be used in clinical settings in cirrhosis in which plasma expansion would reverse some of the decreased effective arterial blood volume, or prevent its iatrogenic (i.e., paracenteses) or spontaneous worsening (spontaneous bacterial peritonitis). However, apart from the issue of transmission of prion agents, which may become an important issue in clinical risk management of the use of albumin in the future, the problem with albumin is its expense. Every effort must thus be made to definitely prove albumin is always the best colloid for all clinical settings in cirrhosis. Further randomized trials are justified. PMID- 14563192 TI - Use of albumin in the management of patients with decompensated cirrhosis. An independent verdict. AB - The use of intravenous albumin in cirrhosis has been reactivated during the last two decades. During this period several investigations have shown that albumin (1) prevents circulatory dysfunction in patients with massive ascites treated by paracentesis, (2) prevents circulatory dysfunction and type-1 HRS and increases survival in patients with SBP, and (3) in association with vasoconstrictors normalizes circulatory function and serum creatinine and increases survival in patients with type-1 HRS. Indications 2 and 3 are clear. There is discussion, however, regarding indication number 1. Although no significant differences in survival have been observed in trials comparing patients treated by paracentesis with and without albumin, in none of these studies was survival an end-point of the trial. In contrast, there is evidence that paracentesis-induced circulatory dysfunction is associated with a bad outcome. In consequence, although further studies on this indication are clearly required, with the current data it is advisable to use albumin as a plasma expander in patients with massive ascites treated by paracentesis. PMID- 14563193 TI - Macroenzyme: do not forget liver function tests. PMID- 14563195 TI - Delivery of midinfrared (6 to 7-microm) laser radiation in a liquid environment using infrared-transmitting optical fibers. AB - Ablation at wavelengths near lambda = 6.45 microm results in tissue ablation with minimal collateral damage (< 40 microm) yet yields a high ablation rate that is useful for human surgery. However, delivery of this wavelength has been limited to that in air and thus to applications in which the target tissue can be readily exposed. The goal of this study is to investigate the potential of a pulsed infrared laser at lambda = 6.45 microm for noncontact ablation in a liquid environment. To this end we investigated fiber delivery in combination with the use of infrared transparent liquids. Transmission characteristics and damage thresholds for two types of fiber materials (silver halide and arsenic sulfide), for high-power pulsed laser radiation were determined using the Mark III free electron laser. Both fibers had comparable bulk losses (0.54 dB/m and 0.62 dB/m, respectively) while the arsenic sulfide fibers showed more coupling losses (37 versus 27%). Damage thresholds were higher in arsenic sulfide fibers than in silver halide fibers (1.12 GW/cm2 versus 0.54 GW/cm2), but both fibers were sufficient to deliver radiant exposures well above the ablation threshold in tissue. Seven different perfluorocarbon liquids (PFCLs), known for their transparency at lambda = 2.94 microm, were investigated and their optical transmission was determined using Fourier transform infrared and direct Beer's law measurements. All of the PFCLs tested had similar values for an absorption coefficient mu(a) at a given wavelength (mu(a) = 0.05 mm(-1) at lambda = 2.94 microm and mu(a) is approximately 3 mm(-1) at lambda = 6.45 microm). Pump-probe imaging showed the ablation sequence (lambda = 6.45 microm) at the fiber tip in a water environment, which revealed a fast expanding and collapsing bubble. In contrast, the volatile PF-5060 showed no fast bubble expansion and collapse, but rather formation of nontransient gas bubbles. Perfluorodecalin did not show any bubble formation at the radiant exposures used. It was shown that using the lambda = 6.45 microm wavelength delivered via fiber optics in combination with perfluorodecalin allows a noncontact laser surgical procedure. Deeper structures, however, are effectively shielded because the radiant exposure of the beam will fall below the ablation threshold owing to the absorption by perfluorodecalin. This may optimize the efficacy and safety of laser-based vitreoretinal surgery. PMID- 14563196 TI - Wound healing of 6.45-microm free electron laser skin incisions with heat conducting templates. AB - We have previously shown a reduction in lateral thermal damage with acute studies of skin incisions made in vitro using heat-conducting templates. Here we examined the wound-healing response to laser incisions with heat-conducting templates and explored the use of an optically transparent template with the free electron laser (FEL) at 6.45 microm. First we evaluated the effects of a sapphire heat conducting template on the lateral thermal damage of FEL incisions using in vitro human skin samples. Next we compared wound tensile strength and histological scoring of the healing of incisions created on the dorsal pelts of live rats with the FEL utilizing metal and sapphire heat-conducting templates and scalpel incisions. The animals were euthanized and the wounds were analyzed at postoperative days 7, 14, and 21. The depth and lateral thermal damage of FEL incisions on in vitro human skin were significantly reduced with the sapphire heat-conducting template. Nonstatistically significant differences in wound tensile strengths and histological scoring of wound healing were noted at days 7 and 14. By day 21, all of the incisions appeared similar. When the data from days 7 and 14 were combined, statistically significant differences were found for each of the templates (except the histological evaluation with the aluminum template) and the scalpel compared with laser incisions made without using a template. The use of metal or sapphire heat-conducting templates reduced the wound-healing delay of laser incisions seen at postoperative days 7 and 14. PMID- 14563197 TI - Quantification of the horny layer using tape stripping and microscopic techniques. AB - Tape stripping is a method well suited for studying the penetration of topically applied substances into the upper part of the skin, the stratum corneum (SC). The amount of topically applied substances removed by each tape strip can be determined by common analytical methods. These amounts have to be correlated with their location within the horny layer in order to determine penetration profiles. Therefore the amount of SC on each removed strip must be determined. In the study reported, this amount was derived from the covering density of the SC cells, the corneocytes, on tape strips using classic microscopic techniques and laser scanning microscopy. The amount of corneocytes determined by both microscopic methods shows a good correlation (R2 +/- S.D.= 0.95 +/- 0.02) with the pseudoabsorption of the corneocytes, a method usually used to determine the SC amount on the removed strips. These measurements require the application of a special spectrometer, whereas the covering density can easily be determined using widely available microscopic equipment. The penetration of a typical UV filter, butyl methoxydibenzoylmethane, applied at two different formulations, was studied using the microscopic method developed. The calculated penetration profiles show that the applied formulation influences the amount of corneocytes on the removed strips. PMID- 14563198 TI - Polarized light propagation through scattering media: time-resolved Monte Carlo simulations and experiments. AB - A study of polarized light transmitted through randomly scattering media of a polystyrene-microsphere solution is described. Temporal profiles of the Stokes vectors and the degree of polarization are measured experimentally and calculated theoretically based on a Monte Carlo technique. The experimental results match the theoretical results well, which demonstrates that the time-resolved Monte Carlo technique is a powerful tool that can contribute to the understanding of polarization propagation in biological tissue. Analysis based on the Stokes Mueller formalism and the Mie theory shows that the first scattering event determines the major spatial patterns of the transmitted Stokes vectors. When an area detected at the output surface of a turbid medium is circularly symmetrical about the incident beam, the temporal profile of the transmitted light is independent of the incident polarization state. A linear relationship between the average order of the scatters and the light propagation time can be used to explain the exponential decay of the degree of polarization of transmitted light. PMID- 14563199 TI - Small-volume frequency-domain oximetry: phantom experiments and first in vivo results. AB - We describe a new method to determine the oxygen saturation and the total hemoglobin content of tissue in vivo absolutely at small source-detector separations (<10 mm). Phase and mean intensity of modulated laser light of various wavelengths was measured at several predetermined source-detector separations in the frequency domain. From these measured quantities, the absorption coefficient was derived using the modified time-integrated microscopic Beer-Lambert law (MBL). In addition, the interaction volume of the photons was determined using a multi-layer Monte-Carlo model of human skin. To evaluate the method, we employed homogenous solid phantoms (consisting of TiO2 particles embedded in resin) with mean scattering and absorbing properties comparable to those of human skin. Furthermore, in vivo measurements were performed in a healthy volunteer to demonstrate that the technique is applicable for the determination of the oxygen saturation and the total hemoglobin content in the skin in vivo. The proposed technique is especially suited for the on-line determination of the oxygen saturation and total hemoglobin content in applications where small applicators are required (e.g., fetal oxygen monitoring sub partu). PMID- 14563200 TI - Spatiotemporal characteristics of cerebral blood volume changes in rat somatosensory cortex evoked by sciatic nerve stimulation and obtained by optical imaging. AB - The spatiotemporal characteristics of changes in cerebral blood volume associated with neuronal activity were investigated in the hindlimb somatosensory cortex of alpha-chloralose-urethane anesthetized rats (n=10) with optical imaging at 570 nm through a thinned skull. Activation of the cortex was carried out by electrical stimulation of the contralateral sciatic nerve with 5-Hz, 0.3-V pulses (0.5 ms) for 2 s. The stimulation evoked a monophasic decrease in optical reflectance at the cortical parenchyma and arterial sites soon after the onset of stimulation, whereas no similar response was observed at vein compartments. The optical signal changes reached 10% of the peak response 0.70 +/- 0.32 s after the start of stimulation, and no significant time lag in this 10% start latency time was observed between the response at the cortical parenchyma and artery compartments. The decrease in optical reflectance reached a peak (0.25 +/- 0.047%) 2.66 +/- 0.61 s after stimulus onset at parenchymal sites, which is 0.40 +/- 0.20 s earlier (P<0.05) than that at arterial sites (0.50 +/- 0.068% 3.06 +/- 0.70 s). Varying the locations within the cortical parenchyma and arterial compartments did not significantly affect the temporal characteristics of the evoked signal. These results suggest that stimulation of the sciatic nerve evokes an increase in local blood volume in both capillaries (cortical parenchyma) and arterioles soon after the onset of a stimulus, but the blood volume increase evoked in capillaries could not be entirely accounted for by the dilation of arterioles. PMID- 14563201 TI - Optical imaging of metastatic tumors using a folate-targeted fluorescent probe. AB - We describe the use of a tumor targeting ligand, the vitamin folic acid, to deliver an attached fluorescent probe to both primary and metastatic tumors overexpressing the folate receptor. Upon laser excitation, derived images of normal tissues generally show little or no fluorescence, whereas images of folate receptor-expressing tumors display bright fluorescence that can be easily distinguished from adjacent normal tissue. Furthermore, metastatic tumor loci of submillimeter size can also be visualized without the aid of image processing or enhancement. The sharp distinction between tumor and normal tissues provided by this technique could find application in the localization and resection of tumor tissue during surgery or in the enhanced endoscopic detection and staging of cancers. PMID- 14563202 TI - Correlation of quantitative light-induced fluorescence and optical coherence tomography applied for detection and quantification of early dental caries. AB - Fluorescence loss in enamel following demineralization has been correlated with the amount of mineral lost during the demineralization. The correlation between fluorescence loss measured by quantitative light-induced fluorescence (QLF) and the reflectivity loss measured by a versatile en face optical coherence tomography (OCT) system was investigated in a demineralization process to produce artificial dental caries. We used an OCT system that can collect A-scans (reflectivity versus depth), B-scans (longitudinal images), and C-scans (en face images). The power to the sample was 250 microW, the wavelength lambda = 850 nm, and the depth resolution in air 16 microm. A-scans, which show the profile of the reflectivity versus the depth of penetration into the tooth tissue, were used for quantitative analysis of the reflectivity loss. The results have shown that both the fluorescence radiance and reflectivity of the enamel decrease with increasing de- mineralization time. A linear correlation was observed between the percentage of fluorescence loss measured by QLF and the percentage of reflectivity loss measured by OCT. It was concluded that the decrease in reflectivity of the enamel during demineralization, measured by OCT, could be related to the amount of mineral lost during the de- mineralization process. PMID- 14563203 TI - Hand-held arthroscopic optical coherence tomography for in vivo high-resolution imaging of articular cartilage. AB - We describe a novel hand-held polarization optical coherence tomographic (OCT) probe that can be inserted into mammalian joints to permit real-time cross sectional imaging of articular cartilage. The transverse and axial resolutions of the arthroscopic OCT device are roughly 17 and 10 microm, respectively. Two dimensional cross-sectional images of cartilage tissue with 500 x 1000 pixels covering an area 6 mm in length and 2.8 mm in depth can be acquired at nearly five frames/s and with over 100 dB of dynamic range. Design of an OCT as a hand held device capable of providing such an optical biopsy of articular cartilage allows eventual in vivo detection of microstructural changes in articular cartilage that are not apparent using conventional arthroscopic cameras. The OCT probe can be easily incorporated in a conventional arthroscope for cartilage site guidance. The optical arrangement in the OCT scope minimizes specular back reflection of the probe end face and absorption of body fluid in the path and ensures in-focus OCT imaging when it is in contact with the cartilage specimen to be examined. Successful application of in vivo arthroscopy to porcine articular cartilage demonstrates sufficient resolution and practicality for use in human joints. PMID- 14563204 TI - Axial and peripheral eye length measured with optical low coherence reflectometry. AB - An optical low-coherence reflectometer (OLCR device) is described that allows the precise and noncontact measurement of eye length. The device measures eye length both on-axis and off-axis, thus enabling the determination of eye shape, an ocular parameter thought to be important in the development of refractive error. It is essential for several applications in ophthalmology and vision science. This improved OLCR device operates using a single-beam interferometer with a beam deflection mechanism that allows the precise measurement of eye length along the visual axis and within 15 deg horizontally and vertically from the fovea. The validity of this instrument and its revised software is evaluated by measuring the reproducibility of axial length results in an adult eye and an artificial eye, and by correlating axial eye length measured in a group of ten adult eyes with axial eye length obtained with A-scan ultrasound in the same eyes. The precision obtained with adult subjects is compared with that obtained with children. PMID- 14563205 TI - Simple device for quantifying the influence of halos after lasik surgery. AB - We present a simple device (called a halometer) to detect and quantify the phenomenon of halos after certain surgical procedures, such as refractive surgery. The task of the subject consists basically of discriminating, after dark adaptation, a small light source around a central high-luminance stimulus. The device, which is connected to a computer, provides a disturbance index to measure the effect of halos on the observer's vision. Tested with normal subjects and patients after LASIK (laser in situ keratomileusis) surgery, this apparatus proved sufficiently sensitive to quantify the halos in subjects who had undergone surgery. PMID- 14563208 TI - A common mechanism for the regulation of vesicular SNAREs on phospholipid membranes. AB - The SNARE (soluble N -ethylmaleimide-sensitive fusion protein attachment protein receptor) family of proteins is essential for membrane fusion in intracellular traffic in eukaryotic organisms. v-SNAREs (vesicular SNAREs) must engage target SNAREs in the opposing membrane to form the fusogenic SNARE complex. Temporal and spatial control of membrane fusion is important for many aspects of cell physiology and may involve the regulation of the SNAREs resident on intracellular membranes. Here we show that the v-SNARE synaptobrevin 2, also known as VAMP (vesicle-associated membrane protein) 2, is restricted from forming the SNARE complex in chromaffin granules from adrenal medullae to the same degree as in brain-purified synaptic vesicles. Our analysis indicates that the previously reported synaptophysin-synaptobrevin interaction is not likely to be involved in regulation of the v-SNARE. Indeed, the restriction can be reproduced for two distinct v-SNARE homologues, synaptobrevin 2 and cellubrevin/VAMP3, by reconstituting them in pure liposomal membranes. Overall, our data uncover a common mechanism for the control of SNARE engagement where intact phospholipid membranes rather than proteins down-regulate vesicular SNAREs in different cellular organelles. PMID- 14563207 TI - Distinct roles for insulin and insulin-like growth factor-1 receptors in pancreatic beta-cell glucose sensing revealed by RNA silencing. AB - The importance of the insulin receptor (IR) and the insulin-like growth factor-1 receptor (IGF-1R) for glucose-regulated insulin secretion and gene expression in pancreatic islet beta-cells is at present unresolved. Here, we have used small interfering RNAs (siRNAs) to silence the expression of each receptor selectively in clonal MIN6 beta-cells. Reduction of IR levels by >90% completely inhibited glucose (30 mM compared with 3 mM)-induced insulin secretion, but had no effect on depolarization-stimulated secretion. IR depletion also blocked the accumulation of preproinsulin (PPI), pancreatic duodenum homoeobox-1 (PDX-1) and glucokinase (GK) mRNAs at elevated glucose concentrations, as assessed by quantitative real-time PCR analysis (TaqMan). Similarly, depletion of IGF-1R inhibited glucose-induced insulin secretion but, in contrast with the effects of IR silencing, had little impact on the regulation of gene expression by glucose. Moreover, loss of IGF-1R, but not IR, markedly inhibited glucose-stimulated increases in cytosolic and mitochondrial ATP, suggesting a role for IGF-1R in the maintenance of oxidative metabolism and in the generation of mitochondrial coupling factors. RNA silencing thus represents a useful tool for the efficient and selective inactivation of receptor tyrosine kinases in isolated beta-cells. By inhibiting glucose-stimulated insulin secretion through the inactivation of IGF-1R, this approach also demonstrates the existence of insulin-independent mechanisms whereby elevated glucose concentrations regulate PPI, PDX-1 and GK gene expression in beta-cells. PMID- 14563209 TI - Engineering peroxidase activity in myoglobin: the haem cavity structure and peroxide activation in the T67R/S92D mutant and its derivative reconstituted with protohaemin-l-histidine. AB - Atomic co-ordinates and structure factors for the T67R/S92D metMbCN mutant have been deposited with the Protein Data Bank, under accession codes 1h1x and r1h1xsf, respectively. Protein engineering and cofactor replacement have been employed as tools to introduce/modulate peroxidase activity in sperm whale Mb (myoglobin). Based on the rationale that haem peroxidase active sites are characterized by specific charged residues, the Mb haem crevice has been modified to host a haem-distalpropionate Arg residue and a proximal Asp, yielding the T67R/S92D Mb mutant. To code extra conformational mobility around the haem, and to increase the peroxidase catalytic efficiency, the T67R/S92D Mb mutant has been subsequently reconstituted with protohaem-L-histidine methyl ester, yielding a stable derivative, T67R/S92D Mb-H. The crystal structure of T67R/S92D cyano-metMb (1.4 A resolution; R factor, 0.12) highlights a regular haem-cyanide binding mode, and the role for the mutated residues in affecting the haem propionates as well as the neighbouring water structure. The conformational disorder of the haem propionate-7 is evidenced by the NMR spectrum of the mutant. Ligand-binding studies show that the iron(III) centres of T67R/S92D Mb, and especially of T67R/S92D Mb-H, exhibit higher affinity for azide and imidazole than wild-type Mb. In addition, both protein derivatives react faster than wild-type Mb with hydrogen peroxide, showing higher peroxidase-like activity towards phenolic substrates. The catalytic efficiency of T67R/S92D Mb-H in these reactions is the highest so far reported for Mb derivatives. A model for the protein-substrate interaction is deduced based on the crystal structure and on the NMR spectra of protein-phenol complexes. PMID- 14563210 TI - Arf-1 (ADP-ribosylation factor-1) is involved in the activation of a mammalian Na+-selective current. AB - Stimulation of mammalian cells often results in an increase in the intracellular Na(+) concentration, brought about by Na(+) influx into the cell via Na(+) permeable ion channels. In some cell types, particularly renal epithelia and mast cells, non-hydrolysable analogues of GTP, such as GTP[S] (guanosine 5'-[gamma thio]triphosphate), activate a non-voltage-activated Na(+)-selective current. We have carried out whole-cell patch-clamp experiments to examine how GTP[S] activates the Na(+) current in a rat mast cell line. The ability of GTP[S] to activate Na(+) influx was prevented by including GTP in the pipette solution, indicating the involvement of small G-proteins. Brefeldin A and Arf-1-(2-17), inhibitors of Arf-1 (ADP-ribosylation factor-1) proteins, suppressed the activation of Na(+) entry by GTP[S]. However, non-active succinylated Arf-1-(2 17) or an N-terminal myristoylated peptide directed towards Arf-5 were ineffective. Arf proteins modulate the cytoskeleton, and disruption of the cytoskeleton with cytochalasin D or its stabilization with phalloidin impaired the development of the Na(+) current. Disaggregation of microtubules was without effect. Dialysis with cAMP or inhibition of cAMP phosphodiesterase with caffeine both decreased the extent of Na(+) entry, and this was not prevented by pre treatment with broad-spectrum protein kinase inhibitors. Collectively, our results suggest that the mechanism of activation of a mammalian non-voltage activated Na(+)-selective current requires an Arf small G-protein, most probably Arf-1. PMID- 14563211 TI - Computer-aided tissue engineering: overview, scope and challenges. AB - Advances in computer-aided technology and its application with biology, engineering and information science to tissue engineering have evolved a new field of computer-aided tissue engineering (CATE). This emerging field encompasses computer-aided design (CAD), image processing, manufacturing and solid free-form fabrication (SFF) for modelling, designing, simulation and manufacturing of biological tissue and organ substitutes. The present Review describes some salient advances in this field, particularly in computer-aided tissue modeling, computer-aided tissue informatics and computer-aided tissue scaffold design and fabrication. Methodologies of development of CATE modelling from high-resolution non-invasive imaging and image-based three-dimensional reconstruction, and various reconstructive techniques for CAD-based tissue modelling generation will be described. The latest development in SFF to tissue engineering and a framework of bio-blueprint modelling for three-dimensional cell and organ printing will also be introduced. PMID- 14563212 TI - The TROVE module: a common element in Telomerase, Ro and Vault ribonucleoproteins. AB - BACKGROUND: Ribonucleoproteins carry out a variety of important tasks in the cell. In this study we show that a number of these contain a novel module, that we speculate mediates RNA-binding. RESULTS: The TROVE module--Telomerase, Ro and Vault module--is found in TEP1 and Ro60 the protein components of three ribonucleoprotein particles. This novel module, consisting of one or more domains, may be involved in binding the RNA components of the three RNPs, which are telomerase RNA, Y RNA and vault RNA. A second conserved region in these proteins is shown to be a member of the vWA domain family. The vWA domain in TEP1 is closely related to the previously recognised vWA domain in VPARP a second component of the vault particle. This vWA domain may mediate interactions between these vault components or bind as yet unidentified components of the RNPs. CONCLUSIONS: This work suggests that a number of ribonucleoprotein components use a common RNA-binding module. The TROVE module is also found in bacterial ribonucleoproteins suggesting an ancient origin for these ribonucleoproteins. PMID- 14563213 TI - Topographical expression of class IA and class II phosphoinositide 3-kinase enzymes in normal human tissues is consistent with a role in differentiation. AB - BACKGROUND: Growth factor, cytokine and chemokine-induced activation of PI3K enzymes constitutes the start of a complex signalling cascade, which ultimately mediates cellular activities such as proliferation, differentiation, chemotaxis, survival, trafficking, and glucose homeostasis. The PI3K enzyme family is divided into 3 classes; class I (subdivided into IA and IB), class II (PI3K-C2alpha, PI3K C2beta and PI3K-C2gamma) and class III PI3K. Expression of these enzymes in human tissue has not been clearly defined. METHODS: In this study, we analysed the immunohistochemical topographical expression profile of class IA (anti-p85 adaptor) and class II PI3K (PI3K-C2alpha and PI3K-C2beta) enzymes in 104 formalin fixed, paraffin embedded normal adult human (age 33-71 years, median 44 years) tissue specimens including those from the gastrointestinal, genitourinary, hepatobiliary, endocrine, integument and lymphoid systems. Antibody specificity was verified by Western blotting of cell lysates and peptide blocking studies. Immunohistochemistry intensity was scored from undetectable to strong. RESULTS: PI3K enzymes were expressed in selected cell populations of epithelial or mesenchymal origin. Columnar epithelium and transitional epithelia were reactive but mucous secreting and stratified squamous epithelia were not. Mesenchymal elements (smooth muscle and endothelial cells) and glomerular epithelium were only expressed PI3K-C2alpha while ganglion cells expressed p85 and PI3K-C2beta. All three enzymes were detected in macrophages, which served as an internal positive control. None of the three PI3K isozymes was detected in the stem cell/progenitor compartments or in B lymphocyte aggregates. CONCLUSIONS: Taken together, these data suggest that PI3K enzyme distribution is not ubiquitous but expressed selectively in fully differentiated, non-proliferating cells. Identification of the normal in vivo expression pattern of class IA and class II PI3K paves the way for further analyses which will clarify the role played by these enzymes in inflammatory, neoplastic and other human disease conditions. PMID- 14563214 TI - Identification of roots from grass swards using PCR-RFLP and FFLP of the plastid trnL (UAA) intron. AB - BACKGROUND: The specific associations between plant roots and the soil microbial community are key to understanding nutrient cycling in grasslands, but grass roots can be difficult to identify using morphology alone. A molecular technique to identify plant species from root DNA would greatly facilitate investigations of the root rhizosphere. RESULTS: We show that trnL PCR product length heterogeneity and a maximum of two restriction digests can separate 14 common grassland species. The RFLP key was used to identify root fragments at least to genus level in a field study of upland grassland community diversity. Roots which could not be matched to known types were putatively identified by comparison of the nuclear ribosomal ITS sequence to the GenBank database. Ten taxa were identified among almost 600 root fragments. Additionally, we have employed capillary electrophoresis of fluorescent trnL PCR products (fluorescent fragment length polymorphism, FFLP) to discriminate all taxa identified at the field site. CONCLUSION: We have developed a molecular database for the identification of some common grassland species based on PCR-RFLP of the plastid transfer RNA leucine (trnL) UAA gene intron. This technique will allow fine-scale studies of the rhizosphere, where root identification by morphology is unrealistic and high throughput is desirable. PMID- 14563215 TI - Insomnia is a frequent finding in adults with Asperger syndrome. AB - BACKGROUND: Asperger syndrome (AS) is a neurodevelopmental disorder belonging to autism spectrum disorders with prevalence rate of 0,35% in school-age children. It has been most extensively studied in childhood while there is scarcity of reports concerning adulthood of AS subjects despite the lifelong nature of this syndrome. In children with Asperger syndrome the initiation and continuity of sleep is disturbed because of the neuropsychiatric deficits inherent of AS. It is probable that sleep difficulties are present in adulthood as well. Our hypothesis was that adults with AS suffer from difficulty in initiating and maintaining sleep and nonrestorative sleep (insomnia). METHODS: 20 AS without medication were compared with 10 healthy controls devoid of neuropsychiatric anamnesis. Clinical examination, blood test battery and head MRI excluded confounding somatic illnesses. Structured psychiatric interview for axis-I and axis-II disorders were given to both groups as well as Beck Depression Inventory and Wechsler adult intelligence scale, revised version.Sleep quality was assessed with sleep questionnaire, sleep diary during 6 consecutive days and description of possible sleep problems by the participants own words was requested. RESULTS: compared with controls and with normative values of good sleep, AS adults had frequent insomnia. In sleep questionnaire 90% (18/20), in sleep diary 75% (15/20) and in free description 85% (17/20) displayed insomnia. There was a substantial psychiatric comorbidity with only 4 AS subject devoid of other axis-I or axis-II disorders besides AS. Also these persons displayed insomnia. It can be noted that the distribution of psychiatric diagnoses in AS subjects was virtually similar to that found among patient with chronic insomnia. CONCLUSIONS: the neuropsychiatric deficits inherent of AS predispose both to insomnia and to anxiety and mood disorders. Therefore a careful assessment of sleep quality should be an integral part of the treatment plan in these individuals. Conversely, when assessing adults with chronic insomnia the possibility of autism spectrum disorders as one of the potential causes of this condition should be kept in mind. PMID- 14563216 TI - Recovery of renal function in dialysis patients. AB - BACKGROUND: Although recovery of renal functions in dialysis dependent patients is estimated to be greater than 1%, there are no indicators that actually suggest such revival of renal function. Residual renal function in dialysis patients is unreliable and seldom followed. Therefore renal recovery (RR) in dialysis dependent patients may remain unnoticed. We present a group of dialysis dependent patients who regained their renal functions. The aim of this project is to determine any indicators that may identify the recovery of renal functions in dialysis dependent patients. METHODS: All the discharges from the chronic dialysis facilities were identified. Among these discharges deaths, transplants, voluntary withdrawals and transfers either to another modality or another dialysis facility were excluded in order to isolate the patients with RR. The dialysis flow sheets and medical records of these patients were subsequently reviewed. RESULTS: Eight patients with a mean age of 53.8 +/- 6.7 years (+/- SEM) were found to have RR. Dialysis was initiated due to uremic symptoms in 6 patients and fluid overload in the remaining two. The patients remained dialysis dependent for 11.1 +/- 4.2 months. All these patients had good urine output and 7 had symptoms related to dialysis. Their mean pre-initiation creatinine and BUN levels were 5.21 +/- 0.6 mg/dl and 72.12 +/- 11.12 mg/dl, respectively. Upon discontinuation, they remained dialysis free for 19.75 +/- 5.97 months. The mean creatinine and BUN levels after cessation of dialysis were 2.85 +/- 0.57 mg/dl and 29.62 +/- 5.26 mg/dl, respectively, while the mean creatinine clearance calculated by 24-hour urine collection was 29.75 +/- 4.78 ml/min. One patient died due to HIV complications. One patient resumed dialysis after nine months. Remaining continue to enjoy a dialysis free life. CONCLUSION: RR must be considered in patients with good urine output and unresolved acute renal failure. Dialysis intolerance may be an indicator of RR among such patients. PMID- 14563217 TI - Determination of decimal reduction time (D value) of chemical agents used in hospitals for disinfection purposes. AB - BACKGROUND: Prior to the selection of disinfectants for low, intermediate and high (sterilizing) levels, the decimal reduction time, D-value, for the most common and persistent bacteria identified at a health care facility should be determined. METHODS: The D-value was determined by inoculating 100 mL of disinfecting solution with 1 mL of a bacterial suspension (10(4)-10(5) CFU/mL for vegetative and spore forms). At regular intervals, 1 mL aliquots of this mixture were transferred to 8 mL of growth media containing a neutralizing agent, and incubated at optimal conditions for the microorganism. RESULTS: The highest D values for various bacteria were determined for the following solutions: (i) 0.1% sodium dichloroisocyanurate (pH 7.0)--E. coli and A. calcoaceticus (D = 5.9 min); (ii) sodium hypochlorite (pH 7.0) at 0.025% for B. stearothermophilus (D = 24 min), E. coli and E. cloacae (D = 7.5 min); at 0.05% for B. stearothermophilus (D = 9.4 min) and E. coli (D = 6.1 min) and 0.1% for B. stearothermophilus (D = 3.5 min) and B. subtilis (D = 3.2 min); (iii) 2.0% glutaraldehyde (pH 7.4)--B. stearothermophilus, B. subtilis (D = 25 min) and E. coli (D = 7.1 min); (iv) 0.5% formaldehyde (pH 6.5)--B. subtilis (D = 11.8 min), B. stearothermophilus (D = 10.9 min) and A. calcoaceticus (D = 5.2 min); (v) 2.0% chlorhexidine (pH 6.2)--B. stearothermophilus (D = 9.1 min), and at 0.4% for E. cloacae (D = 8.3 min); (vi) 1.0% Minncare (peracetic acid and hydrogen peroxide, pH 2.3)--B. stearothermophilus (D = 9.1 min) and E. coli (D = 6.7 min). CONCLUSIONS: The suspension studies were an indication of the disinfectant efficacy on a surface. The data in this study reflect the formulations used and may vary from product to product. The expected effectiveness from the studied formulations showed that the tested agents can be recommended for surface disinfection as stated in present guidelines and emphasizes the importance and need to develop routine and novel programs to evaluate product utility. PMID- 14563218 TI - Anorectal motility in patients with achalasia of the esophagus: recognition of an esophago-rectal syndrome. AB - BACKGROUND: During my study of constipation, I encountered patients who had achalasia of the esophagus (AE) as well. The possibility of an existing relationship between the 2 conditions was studied. METHOD: Investigations to study the anorectal motility in 9 AE patients included: the intestinal transit time, anorectal manometry, rectoanal inhibitory reflex, defecography and electromyography (EMG) of external anal sphincter and levator ani muscle. Anorectal biopsy was done. The study comprised 8 healthy volunteers as controls. RESULTS: 6/9 AE patients had constipation presenting as strainodynia (excessive prolonged straining at stool). Rectocele was present in 4 of them. The 6 constipated patients showed significantly high rectal neck pressure (p < 0.05), absent rectoanal inhibitory reflex and aganglionosis in the anorectal biopsy. The EMG revealed diminished activity in 4 of the 6 constipated patients. The remaining 3 patients with AE had normal anorectal function. Heller's myotomy with Nissen's fundoplication improved the dysphagia, but not the constipation which was, however, relieved after performance of anorectal myectomy. CONCLUSION: The high incidence of constipation with AE postulates a relationship between the 2 conditions. Both have the same pathologic lesion which is aganglionosis. This study is preliminary and requires further studies on a larger number of patients. PMID- 14563221 TI - Pediatric invasive pneumococcal disease in a teaching hospital in Bangkok. AB - BACKGROUND: Increased problems with drug-resistant Streptococcus pneumoniae (SP) and the dearth of epidemiologic and clinical information on invasive pneumococcal disease in children in Asia formed the basis for this study. METHODS: A periodic retrospective review of the records of 0-15-year-old patients was conducted at a teaching hospital in Bangkok, during 1971-2000. RESULTS: Infections with penicillin-non-susceptible SP (PNSSP) strains rapidly increased after they first appeared in 1988, and they accounted for 71% (29/41) of the total cases during 1996-2000. Of 137 patients, 74% were <60 months old, and 66% had an underlying condition. Infections included: bacteremia without focus 51; pneumonia 38; meningitis 35; peritonitis 13; and bone/joint infection 2. Two patients had two foci of infection. Eight of 10 episodes in patients with AIDS were bacteremic pneumonia. Median ages (range) in months for patients with and without an underlying condition were 24 (1-174) and 10 (0-160); and for the patients without an underlying condition they were: pneumonia 23 (4-156); bacteremia without focus 12 (0-160); and meningitis 7 (2-156). Case-fatality rates were 18% and 2% for patients with and without an underlying condition. The study also examined factors associated with PNSSP infection and death. During 1991-2000, 74% (43/58) of the total cases occurred from November to April, which are dry months. CONCLUSIONS: This study population contained a high proportion with both an underlying condition and infection with PNSSP, and a moderately low proportion with bacteremia without focus. The disease was two to three times more common in dry months than in rainy months. PMID- 14563219 TI - Mitochondrial DNA transit between West Asia and North Africa inferred from U6 phylogeography. AB - BACKGROUND: World-wide phylogeographic distribution of human complete mitochondrial DNA sequences suggested a West Asian origin for the autochthonous North African lineage U6. We report here a more detailed analysis of this lineage, unraveling successive expansions that affected not only Africa but neighboring regions such as the Near East, the Iberian Peninsula and the Canary Islands. RESULTS: Divergence times, geographic origin and expansions of the U6 mitochondrial DNA clade, have been deduced from the analysis of 14 complete U6 sequences, and 56 different haplotypes, characterized by hypervariable segment sequences and RFLPs. CONCLUSIONS: The most probable origin of the proto-U6 lineage was the Near East. Around 30,000 years ago it spread to North Africa where it represents a signature of regional continuity. Subgroup U6a reflects the first African expansion from the Maghrib returning to the east in Paleolithic times. Derivative clade U6a1 signals a posterior movement from East Africa back to the Maghrib and the Near East. This migration coincides with the probable Afroasiatic linguistic expansion. U6b and U6c clades, restricted to West Africa, had more localized expansions. U6b probably reached the Iberian Peninsula during the Capsian diffusion in North Africa. Two autochthonous derivatives of these clades (U6b1 and U6c1) indicate the arrival of North African settlers to the Canarian Archipelago in prehistoric times, most probably due to the Saharan desiccation. The absence of these Canarian lineages nowadays in Africa suggests important demographic movements in the western area of this Continent. PMID- 14563223 TI - Infections with rapidly growing mycobacteria: report of 20 cases. AB - OBJECTIVES: A series of cases infected with rapidly growing mycobacteria was studied to determine the spectrum of disease, antimicrobial susceptibility, treatment, and outcome. METHODS: The cases identified as infections with rapidly growing mycobacteria in Ramathibodi Hospital from January 1993 to December 1999 were retrospectively studied. RESULTS: Most of the cases had no underlying disease. Only two cases were HIV-infected patients. The presenting clinical features were lymphadenitis (seven cases), skin and/or subcutaneous abscess (seven cases), localized eye infection (four cases), pulmonary infection (one case), and chronic otitis media (one case). Four of seven cases with lymphadenitis had Sweet's syndrome, and one had psoriasis as an associated skin manifestation. Anemia was present in five cases, and improved with treatment of the primary disease. The organisms were Mycobacterium chelonae/abscessus group (17 cases) and Mycobacterium fortuitum group (three cases). Susceptibility patterns of the organisms showed susceptibility to amikacin, netilmicin, and imipenem. M. fortuitum group was susceptible to more antibiotics than M. chelonae/abscessus group. The clinical responses corresponded to the antimicrobial susceptibility. Combinations of two or more drugs were used for the medical treatment. Surgical resection was performed where possible, to reduce the load of the organism, especially in cases with very resistant organisms. CONCLUSIONS: Infections with rapidly growing mycobacteria can occur in apparently normal hosts. The clinical syndrome is variable. The pathology is nonspecific. Clinical responses varied, but seemed to correlate with the in vitro susceptibility result. More studies are needed to enable us to deal with this infection effectively. PMID- 14563222 TI - Penicillin susceptibility and molecular characteristics of clinical isolates of Streptococcus pneumoniae at the University of Malaya Medical Center, Kuala Lumpur, Malaysia. AB - OBJECTIVE: To determine the prevalence of penicillin resistance and molecular characteristics of pneumococcal isolates at the University of Malaya Medical Center. METHODS: From March 1999 to July 2000, 100 clinical isolates of Streptococcus pneumoniae were obtained from 93 patients of various ages and from various body sites. The minimum inhibitory concentrations (MICs) for penicillin and ceftriaxone were determined by E test, and results were interpreted according to guidelines recommended by the National Committee for Clinical Laboratory Standards (NCCLS). Fifty isolates were further serotyped, and analyzed by pulsed field gel electrophoresis (PFGE) and polymerase chain reaction/restriction fragment length polymorphism (PCR-RFLP) of the penicillin-binding protein (pbp) 2b and 2x genes. RESULTS: The majority of the isolates were from respiratory sites. Thirty-one isolates showed decreased susceptibility to penicillin (PRSP), and many of these also showed decreased susceptibility to ceftriaxone. Twelve serogroup/types (SGTs) were present, with 19F being the most common. PFGE analysis identified two dominant profiles, consisting mainly of PRSPs that had common serotypes (19F) and pbp gene patterns within their respective groups, although PCR-RFLP analysis showed different patterns of pbp genes among the PRSPs as compared to penicillin-susceptible strains, which had a uniform pattern. CONCLUSION: PRSPs were genetically related as shown by PFGE and serotype. The consistency of pbp gene patterns, observed among many of the PRSPs within their respective PFGE profiles, supported their relatedness as established by PFGE. PMID- 14563224 TI - Prevalence of genital chlamydial infection in symptomatic and asymptomatic Jordanian patients. AB - OBJECTIVE: To detect Chlamydia trachomatis infection among 230 patients, 130 with signs or symptoms associated with urethritis, and 100 asymptomatic patients, attending the Jordan University Hospital urology clinic. METHODS: Routine urine examination and the leukocyte esterase test were done for each patient. C. trachomatis infection was detected using first-void urine specimens and a cryptic plasmid-based PCR technique specific for C. trachomatis. RESULTS: The prevalence of chlamydial infection was 4.6% among symptomatic patients with urethritis. The difference in prevalence was statistically insignificant (P > 0.05) between males and females, as well as in relation to their marital status. Two-thirds of the Chlamydia-positive patients also had urine positive for leukocyte esterase. CONCLUSION: The low prevalence of chlamydial infection in association with urethritis among Jordanian patients might be due to the conservative behavior of the Jordanian society towards free sexuality. PMID- 14563225 TI - A case-control study of influenza vaccine effectiveness among Malaysian pilgrims attending the Haj in Saudi Arabia. AB - OBJECTIVES: To determine influenza vaccine effectiveness against clinically defined influenza-like illness among Malaysian pilgrims attending the Haj in Saudi Arabia. METHODS: During February and March 2000, the authors conducted an unmatched case-control study. Case patients were identified at one of five hotel clinics, while controls were residents of these hotels who had not attended a clinic. RESULTS: Among 820 case patients--84% of whom had received antibiotics- and 600 controls, the adjusted vaccine effectiveness against clinic visits for influenza-like illness was 77% (95% confidence interval: 69, 83), and that against receipt of antibiotics was 66% (95% confidence interval, 54, 75). The vaccine did not prevent clinic visits for non-influenza-like upper respiratory tract illness (adjusted vaccine effectiveness, 20%; 95% confidence interval: -24, 49). CONCLUSIONS: Influenza vaccine was effective in preventing clinic visits for influenza-like illness and antibiotic use. Pilgrims traveling to the Haj in Saudi Arabia should consider influenza vaccination use. PMID- 14563226 TI - Human cytomegalovirus infection during pregnancy and detection of specific T cells by intracellular cytokine staining. AB - OBJECTIVE: The flow cytometric assay was evaluated as a tool for real-time monitoring of human cytomegalovirus (HCMV)-specific cellular immunity in pregnant women. METHODS: We screened for HCMV infection in pregnant women in Sapporo, Japan, during the year 2000, by serologic assays, virus isolation from urine, and PCR to detect DNA in cervical swabs. The frequencies of HCMV-specific CD4+ T cells in pregnant women with serum anti-HCMV IgG antibody were detected by intracellular cytokine (ICC), interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) staining. RESULTS: The levels of intracellular cytokines in pregnant women with serum anti-HCMV IgG antibody were significantly higher than those in women without anti-HCMV IgG antibody (P = 0.011 for IFN-gamma and P = 0.023 for TNF-alpha) but lower than those in non-pregnant women with serum anti HCMV IgG antibody. Frequencies of HCMV-specific CD4+ T cells were higher in infants with symptomatic congenital infection than in infants with asymptomatic perinatal infection. CONCLUSIONS: This ICC assay may reflect immunologic activity against HCMV infection in pregnant women with immunosuppressive conditions. PMID- 14563227 TI - Clinical, histologic and serologic evaluation of patients with acute non-A-E hepatitis in north-eastern Brazil: is it an infectious disease? AB - Non-A-E hepatitis and acute cryptogenic hepatitis are the names given to the disease of patients with clinical hepatitis, but in whom serologic evidence of A E hepatitis has not been found. Over a period of 8 years, we evaluated in Brazil 32 patients who fulfilled the criteria for this diagnosis in order to determine patterns of the clinical illness, laboratory parameters, or histologic features. Each patient was subjected to virologic tests to exclude A-E hepatitis and cytomegalovirus/Epstein-Barr virus infection. Drug-induced hepatitis and autoimmune disease were also excluded. Wilson's disease was excluded in young patients. The course of the disease was clinical/biochemical recovery in 3 months in 25 patients and persistent alanine aminotransferase (ALT) elevation in 7 patients. Three of these had chronic hepatitis, and one had severe fibrosis on liver biopsy. During the acute illness, mean peak ALT was 1267 IU/L, bilirubin was 4.0 mg/dL, and ferritin was 1393 IU/mL. GB virus type C (GBV-C) was found in six patients, and TT virus (TTV) in five patients. We conclude that, in Brazil, non-A-E hepatitis probably originates from still unidentified viruses. The course of the disease and the histologic patterns are similar to those recorded for known viruses. Continuous survey for the specific etiologic agents is needed. PMID- 14563228 TI - Effect of increasing age on the trend of dengue and dengue hemorrhagic fever in Singapore. PMID- 14563229 TI - Malaria: blood slides under treatment. PMID- 14563232 TI - Human mammary abscess caused by Brucella melitensis: a case report. PMID- 14563231 TI - Community-acquired Klebsiella pneumoniae bacteremia with meningitis and endophthalmitis in Italy. PMID- 14563235 TI - [Clinical practice guidelines: an indispensable tool also in osteoporosis]. PMID- 14563236 TI - [Macroprolactin as etiology of hyperprolactinemia. Method for detection and clinical characterization of the entity in 39 patients]. AB - The presence of serum macroprolactin is a relatively frequent situation that can lead to expensive explorations and ineffective treatments. The precipitation with polyethylene glycols permits its detection rapidly, trustworthily, and inexpensively. The objective of the present work has been to assess the incidence of macroprolactin in patients with hyperprolactinemia through its identification with polyethylene glycols, as well as the clinical and radiological findings, and the response in these patients to the treatment. For it the clinical history of all the cases in which there was detected macroprolactinemia, some of them with several years of previous follow-up, was reviewed exhaustively. Of the 1505 patients in which prolactin level was determined, 195 (13%) showed values higher than 40 microg/l. In 39 (20%) of them the presence of macroprolactin was detected. After observing the evolution of these patients, spontaneously or under treatment with dopaminergic agonists, a clear relationship of causality between the presence of macroprolactin and the clinical manifestations was not found, except in a case of galactorrhea. In 27 cases nuclear magnetic resonance was done; in 4 of them a picture was observed compatible with small microadenoma or microcyst and in no patient a macroadenoma was detected. We conclude that the macroprolactinemia is a relatively frequent and easily detected entity through precipitation with polyethylene glycols. It has a limited relation with the clinical findings and there should always be sought other possible causes of this situation. It is an apparently benign situation and its identification can avoid unnecessary explorations and treatments. PMID- 14563237 TI - [Evaluation of the process and the outcomes of care for the diabetic patients in a hospital]. AB - For the purpose of establishing a specific information system in order to identify the diabetic patients looked after in the hospital and evaluate the processes and the clinical outcomes obtained, a prospective descriptive study was designed--in the Corporacio Parc Tauli of Sabadell (Barcelona)--about the diabetic patients cared starting in January 1998, through a clinical registry and other hospital sources of information. Seven hundred and fifty-five patients were identified, and the completeness of the registry was 98%. It was feasible the integration of the registry of admissions with that of the laboratory, that of the discharges from hospital, that of pharmacy and the specific clinical registry. An evaluation of the glycemic control was made in around 90% of the patients, and the registry of the organic explorations was higher than 87%. Although it was not possible in this study, the incorporation of the information from the primary care would complete the follow-up of the patients. PMID- 14563238 TI - [Lung aspergillosis. Role of the corticoids as associated risk factor in the patient with chronic bronchitis]. AB - BASIS: Description of a situation of incidence increase of bronchial secretions with positive cultures for Aspergillus fumigatus, and analysis of the related risk factors in the invasive aspergillosis. METHODS: Between January 1999 and February 2000, a prospective study of the patients was conducted with culture of bronchial secretions and with positive result for A. fumigatus. VARIABLES STUDIED: age, sex, primary diagnosis, type of cultivated sample, clinical interpretation (colonization/infection), probable source (community/nosocomial), situation of the patient after discharge, and risk factors for opportunistic infection. The results were compared among the colonized and infected patients. RESULTS: Fifty-two patients showed positive cultures of bronchial secretions to A. fumigatus, 43 (82.6%) colonized and 9 (17.3%) infected. Cultivated sputum sample on 30 occasions (57.6%) and bronchial aspiration in 22 (42.3%). Median age: 70 years (31-84). Sex: 40 men (76.9%). Probable source of infection/colonization: nosocomial in 18 cases (34.6%), community in 3 (5.7%) and unknown in 31 (59.6%). Mortality: 15 patient colonized (34.8%) and 8 infected (88.8%). Risk factors with statistical significance for invasive infection by A. fumigatus: diagnosis of chronic bronchopathy (COPD) (p=0.007) and treatment with prednisone in dose higher than 60 mg/day (p=0.0005). CONCLUSIONS: The patients with positive culture of bronchial secretions to A. fumigatus with COPD and treatment with prednisone in dose higher than 60 mg/day should be considered with a greater risk for infection by this pathogen. A more restricted use and adequate of the corticoids in these patients, and an early diagnosis and treatment in light of the suspicion of infection by A. fumigatus in patients with COPD, it could imply a reduction of morbidity and mortality. PMID- 14563240 TI - [Immunotherapy in asthma]. PMID- 14563239 TI - [The practice of Internal Medicine in Andalusia follows the evidence-based medicine basic principles]. AB - BASIS: To know the proportion of medical interventions carried out in the Services of Internal Medicine of the public hospitals of Andalusia based on randomized clinical trials. PATIENTS AND METHODS: We have analyzed the primary treatments prescribed in a random sample of 326 patients admitted to these Internal Medicine services during 1998. RESULTS: One hundred and forty-three of the 326 treatments analyzed (43.9%) were based in clinical trials and 135 (41.4%) were interventions unanimously accepted by the medical community without being based in clinical trials. CONCLUSIONS: Most of the primary Andalusian treatments prescribed in the Internal Medicine services are evidence-based. PMID- 14563241 TI - [Treatment of the infections by enterococcus]. PMID- 14563244 TI - [Oral ulcers and erythematopapulous rash in a 17 year-old male]. PMID- 14563243 TI - [Extensive acute intestinal ischemia in a patient with colon carcinoma with local recurrence]. PMID- 14563242 TI - [Treatment of insomnia in dementias]. PMID- 14563245 TI - [Palmoplantar pustulosis, cervicoalgia, and clavicular hyperostosis]. PMID- 14563246 TI - [Postmenopausal osteoporosis. Clinical practice guide]. PMID- 14563247 TI - [Fever of unknown origin and aortic dissection]. PMID- 14563248 TI - [Disease of Kikuchi-Fujimoto and pulmonary tuberculosis, an infrequent association]. PMID- 14563249 TI - [Use of naltrexone in the treatment of alcoholism in Spain]. PMID- 14563250 TI - [Hypervascular pelvic mass: an infrequent cause of digestive hemorrhage]. PMID- 14563251 TI - [Science and scientific literature at the outset of 21st century. Prospects and realities]. PMID- 14563252 TI - [Modifications of the leukocytes in professional cyclists throughout the competition]. AB - In this work we plan to analyze and establish the leukocyte variations due to the effort of the competition, throughout 4 short-term turns (5 days and an average of 750 km each one), in 16 professional cyclists (24.8 2.3 year-old, 71.0 4.5 kg and 179.3 5.0 cm). The hematological parameters analyzed were leukocytes (WBC), neurtrophils (NEUT), lymphocytes (LYM), monocytes (MON), eosinophils (EOS) and basophils (BAS), and the results are presented as the percentage on the total number of leukocytes. The serum level of cortisol was also measured along study time (before and after each one of the 4 turns). The results show that the total number of leukocytes increase significantly in the turns, both in basal conditions (B) and after finishing the turn (F). The NEUT show an ongoing increase in their basal values from the first to the fourth round, but its values at the conclusion of this are significantly lower with respect to the situation of reference of the beginning of the cyclist turn. The LYM in rest upon beginning the turns are kept in similar values during the study period, increasing significantly after the 5 days of competition of each turn. The cortisol, as they are happening the turns, they decline its rates both in rest and after the competition (5 days turns). In conclusion the physical exercise throughout several cyclists turns give rise an increases in the leukocytes that is associated to a progressive decline of the cortisol levels probably because of the conditioning that the cyclist shows throughout the competition. PMID- 14563253 TI - [Basal immunological parameters in a group of retirees]. AB - OBJECTIVES: a) establish the major immunological parameters for clinical use in a group of retirees; b) correlate its levels in relation to gender; c) assess the influence of some specific factors (substance abuse, diseases) on the analyzed parameters. MATERIAL AND METHODS: study period: 1990-1999; sample: 249; 102 men (M); 147 women (W). Median age: 67.03 (4.2) years. Analyzed immunological variables: total leukocytes, lymphocytes B, immunoglobulins (IgG, IgA and IgM), rheumatoid factor, lymphocytes subpopulations (CD4, CD8, ratio), natural killer, complement (C3 and C4) and delayed hypersensitivity tests. TECHNIQUES: flow cytometry (EPICS-Profile II) and Multitest IMC. STATISTICAL ANALYSIS: SPSS version 10.0.Results. Gender influence: leukocytes: M: 6,699.4 (1,615.0); W: 6,105.9 (1,470.5); p < 0,003; lymphocytes B (%): M: 9.4 (5.0); W: 11.3 (4.1); p < 0,003; IgG: M: 1,155.7 (320.0); W: 1,116.1 (257.8); p = 0,28, IgM: M: 112.7 (69.3); W: 136.8 (85.6); p < 0,01; IgA: M: 276.1 (114); W: 254.0 (122); p = 0,15; rheumatoid factor: M: 18.5 (6.6); W: 20.9 (18.8); p = 0,020; CD4 (%): M: 42.2 (9.7); W: 47.3 (9,1); p < 0.001; CD8 (%): M: 30.3 (10,8); W: 25.0 (10.2); p < 0,001; scores: M: 13.2 (7.4); 11m: 10.0 (7.2); p < 0,005. Influence of the substance abuse: smokers; lymphocytes B (%): 8.8 (3.4); No: 10.9 (4.7); p < 0.008; CD8 (%): smokers: 31.8 (13.2); No: 26.2 (9.9); p < 0.003; CD4/CD8 ratio: smokers: 1.6 (0.9); No: 2.0 (1.3); p < 0,05; scores: smokers: 14.3 (6.8); No: 10.8 (7.5); p < 0.02; alcoholism: lymphocytes B (%): 8.7 (2.5). No: 10.8 (4.7); p < 0.001; alcoholism: store: 16.9 (6.7); No: 10.7 (7.3); p < 0,001. Influence of the diseases: diabetes: CD4 (%): 49.4 (12.0); diabetics versus healthy: p = 0,05; CD4/CD8 ratio: 2.6 (2.5); diabetics versus healthy: p = 0,04; EPOC: CD8 (%): 32.9 (16,3); COPD versus healthy: p = 0,07; neoplasias NK (%): 17.1 (21.4); neoplasias versus healthy: p < 0,01. CONCLUSIONS. a) there are differences according to the gender in the parameters of normalcy of some variables; b) smoking and alcoholism alter the immunological test analyzed, and c) some chronic diseases influence the subpopulations of lymphocytes and the cutaneous test of delayed hypersensitivity. PMID- 14563254 TI - [The active and passive voice in the medical literature in Spain and in the United States. A comparative study]. AB - The majority of the manuals on style and scientific writing recommend limiting the use of the passive, preferring the active forms. In order to know the degree of monitoring of this recommendation we have carried out an analysis of a sample of articles of Spanish and United States journals in two different times (1989 and 2001). The use of the active voice is declining in Spain even being almost anecdotal, while in increases in the United States. In both times evaluated the American authors use the active voice more than the Spanish. The possible reasons of this situation are discussed and solutions are suggested. PMID- 14563255 TI - [Prevalence of diabetes treated with drugs in Andalucia. Evolution of the oral antidiabetics and insulin prescription between 1994 and 2000]. AB - CONTEXT: [corrected] Diabetes is an important, complex, expensive, and increasingly frequent disease. The objective of this study was to investigate the diabetes mellitus prevalence and evolution in the Adalusian population between 1994 and 2000. METHODS: We used and indirect method for estimating the rate of prevalence of the disease based on antidiabetic drug consumption data at regional level, using the official billing data, and the annual population data. RESULTS: The prevalence estimated for diabetes mellitus in Andalusia was, 4.41%. This prevalence ranged from 2.47% in 1994 up to 4.41% in 2000. Oral antidiabetics/insulin use ratio was 2.31. The study shows an important variability of the prescription habits in diabetic patients in Andalucia. CONCLUSIONS: The prevalence observed in diabetes mellitus in Andalucia increased between 1994 and 2000. An important variability was observed among the different provinces. PMID- 14563257 TI - [Multiple paraganglioma: careful with surgery!]. AB - INTRODUCTION: Extraadrenal paragangliomas are exceptional tumors. They prevail the carotid, jugulotympanic, and vagal ones. They are frequently multiple and its treatment is controversial in view of the fact that bilateral removal can go with severe morbidity. The case of a patient with bilateral paraganglioma and postoperative baroreflex dysfunction with severe arterial hypertension and hypotension episodes is presented. CLINICAL OBSERVATION: A 23-year-old woman with a diagnosis of left carotid and right vagal paraganglioma by TC and angiography. In January 1999 the left carotid paraganglioma was operated. The patient showed dysphonia, dysphagia, and lingual dysmotility in the postoperative course, with spontaneous improvement after some months. In December 1999, after the removal of the right vagal paraganglioma, the same complications appeared and hypertension crises (230/140), associated with headache, dizziness, and rash, and alternating with severe hypotension episodes (70/50). Blood biochemistry and the levels of cortisol, thyroid hormones, catecholamines, and metabolites were normal. Imaging techniques discarded tumor at another level and the registry of the ambulatory monitoring of blood pressure (AMBP) confirmed an important pressure lability. The neurophysiological study of the autonomous nervous system demonstrated the failure of the fast regulation mechanisms of the blood pressure. With the diagnosis of baroreceptors dysfunction and paralyses of cranial nerves IX, X and XII a treatment with clonidine was started with poor tolerability and incomplete response. DISCUSSION: This case illustrates the treatment difficulties of paragangliomas, especially when they are bilateral, and in which the surgery can go with severe morbidity. Baroreflex dysfunction should be entertained in the differential diagnosis of the extreme pressure lability. PMID- 14563256 TI - [Clinical importance of the CagA and VacA proteins and of the host factores in the development of peptic ulcer in patients infected by Helicobacter pylori]. AB - OBJECTIVE: Demonstrate the role of the virulence factors of Helicobacter pylori, the sex, and tobacco and alcohol use in the development of peptic ulcer in patients infected by H. pylori. PATIENTS AND METHODS: One hundred and seventy four patient with dyspepsia were studied after they went consecutively for gastroscopy. The diagnosis of infection by H. pylori was carried out by culture and/or histology. Through western-blot the presence of specific anti-H. pylori antibodies was determined. The tobacco and alcohol use data were collected. RESULTS: One hundred and twenty-five patients (32 with ulceration, 12 with duodenitis and 81 with functional dyspepsia) showed H. pylori infection with anti bacterium IgG antibodies. The detection of the proteins CagA and VacA, male sex and heavy consumption of alcohol and tobacco were associated with the finding of peptic ulcer. In the multivariate analysis only anti-CagA antibodies (OR: 4.1; 95% CI: 1.1-15.3; p = 0.036), anti-VacA (OR: 3.9; 95% CI: 1.4-10.8; p = 0.009) and male sex (OR: 3.1; 95% CI: 1.2-8.3; p = 0.02) were associated with the ulcerative disease. CONCLUSIONS: The proteins CagA and VacA, and the male sex, contribute independent risk factors for peptic ulcer in patients infected by H. pylori. PMID- 14563258 TI - [Questionnaires of spinal dysfunction: a critical review addressed to the clinician]. PMID- 14563259 TI - [Chronic C virus infection management]. PMID- 14563261 TI - [Dyspnea and intersticial lung pattern]. PMID- 14563260 TI - [Current and future prospects of the treatment of the irritable bowel syndrome]. PMID- 14563262 TI - [A 70-year-old male with repetitive syncope]. PMID- 14563263 TI - [Necrotic lesion in scalp after injury]. PMID- 14563264 TI - [New case of HIV-2 in the Valencia Community]. PMID- 14563265 TI - [Variable common immunodeficiency: description of a case associated with antiphospholipid syndrome]. PMID- 14563266 TI - [General solitary syndrome, are we in agreement with the nomenclature?]. PMID- 14563268 TI - [Reasons for not carrying out stem cell transplantation in patients referred to a transplant unit]. AB - BACKGROUND AND OBJECTIVE: Although indications of stem cell transplantation (SCT) are increasing, a transplant may not be performed in all planned cases for several reasons. Our objective was to investigate the reasons for which SCT was not performed in patients referred to a transplant unit. PATIENTS AND METHOD: Pretransplant data of 129 patients consecutively referred to a transplant unit between December 1999 and November 2002 were collected. Frequency and causes of non transplantation were analyzed. Transplanted and non transplanted patient's characteristics were compared. RESULTS: In 119 out of 129 patients, an autologous SCT was indicated and in 10 of them an allogeneic SCT was planned. Mean (SD) age was 46 (14) years (range, 13-69) and 69 (53.5%) were males. One hundred eighteen patients had malignant hematological diseases and 11 had solid tumours. Sixty-one patients showed complete response and 68 had a partial response. At the time of the analysis, 93 SCT had been performed in 90 (69.8%) patients. Autologous SCT was performed in 81 patients (two SCT in one patient) and allogeneic SCT in 10 (two in one patient). Two (1.5%) patients were still awaiting SCT. An SCT was not performed in 37 (28.7%) patients. Causes of non transplantation included: in 12 (32.5%) cases, relapse and/or progression at the time the SCT had been planned; in 12 (32.5%), delay or change in the therapeutic decision; in 9 (24%), poor mobilization; and in 4 (11%), patient's refusal. When delay in SCT was excluded from the analysis, the frequency of no transplant was 19.4%. In 21 (57%) patients of the non-SCT group, peripheral stem cells were previously collected by apheresis. Both groups (SCT and non-SCT) were comparable regarding patients' and disease characteristics except for a more advanced age in the non-SCT group (51 [12] vs 44 [14] years, p<0.005) and status of the disease at the pretransplant visit. CONCLUSIONS: The frequency of not performing an SCT in patients referred to a SCT unit was considerable. Relapses and disease progression and poor mobilization were the main causes for it. Delay was also a relatively common cause. PMID- 14563269 TI - [Prevalence of cardiovascular risk factors in a cohort of affected by the toxic oil syndrome]. AB - BACKGROUND AND OBJECTIVE: During the first stages of the toxic oil syndrome (TOS), elevations of the blood pressure as well as increases in the cholesterol, glucose and triglycerides levels were reported. Here we analyze these cardiovascular risk factors in the chronic phase of the illness and their distribution according to the severity of the illness. We also compare them with those found in the general population. PATIENTS AND METHOD: We studied a sample of 1,862 individuals aged between 35 and 65 years. A medical examination was performed in each and blood pressure, weight, height, tobacco consumption, cholesterol, glucose and triglycerides levels were measured. RESULTS: The prevalence of high blood pressure (>140/90 mmHg) was 46.1% and the prevalence of diabetes mellitus (>126 mg/dl) was 9.1%. The prevalence of obesity (BMI>30) was 24.9%. 11.8% of patients had hypertriglyceridemia (>200 mg/dl) and 19.8% had hypercholesterolemia (>250 mg/dl). 37.9% were smokers. The standardized prevalence rate (SPR) of high blood pressure was 1.35 (95% CI, 1.28-1.44); tobacco consumption SPR=1.27 (95% CI, 1.20-1.36); hypercholesterolemia SPR=1.10 (95% CI, 1.01-1.21). The prevalence of risk factors was higher among the most seriously affected subjects. CONCLUSIONS: The chronic phase of TOS is characterized by a high prevalence of cardiovascular risk factors, which was significantly higher than that expected in the general population. PMID- 14563270 TI - [Effect of HAART on HCV viral load in the first three years of follow-up]. AB - BACKGROUND AND OBJECTIVE: Highly active antiretroviral therapy (HAART) has been considered to be ineffective in controlling hepatitis C viremia in the first year of treatment. The present study evaluates the effect of prolonged HAART upon HCV VL. PATIENTS AND METHOD: We selected those cases with positive serology for HCV, never been treated with protease inhibitors (PIs) at baseline, which after the introduction of HAART maintained undetectable HIV viral loads (HIV-VL) uninterruptedly for at least three years. Determinations were made of HCV-VL at baseline and after two and three years of HIV-VL suppression. Spearman coefficient and Friedman test were used. RESULTS: Of these patients, 21 satisfied the inclusion criteria. HCV-VL remained without significant changes and with relatively stable levels throughout the three-year follow-up period. However, two patients showed HCV-VL negative conversion after two years, that persisted after three years. CONCLUSIONS: Patients who respond to HAART maintain stable HCV-VL. Some subjects may exhibit HCV RNA clearance in the first two years of treatment, maintaining this situation after three years. PMID- 14563272 TI - [Differential diagnosis between chronic pain and chronic fatigue: a pending matter]. PMID- 14563271 TI - [Human immunodeficiency virus and hepatitis C virus co-infection. Do we know its clinical course in the HAART era?]. PMID- 14563274 TI - [Residual risk of human immunodeficiency virus infection in blood banks. Impact of screening with nucleic acid tests]. AB - In most developed countries of the world, the risk of human immunodeficiency virus (HIV) transmission by transfusion of blood and blood products is extraordinarily small. This level of blood safety has been accomplished by successive refinement in donor screening and testing procedures for the detection and inactivation of different infectious agents in blood and blood products. In USA, the recent introduction of nucleic acid techniques (NAT) in blood banks for the detection of HIV and hepatitis C virus (HCV) has meant a great advance in decreasing the residual risk of HIV/HCV transmission by blood transfusion. In general, after analyzing the first four-years of NATs experience in US blood banks, the introduction of NATs in European blood centers could be considered, since this technique has shown improved output to detect donations from individuals in the very early stages of infection before detectable serologic response has been developed. PMID- 14563273 TI - [Regional anesthesia, thromboprophylaxis and their relationship with the spinal hematoma: Spanish experience]. PMID- 14563275 TI - [Interstitial lung disease]. AB - Molecular investigation into the physiopathology of interstitial lung diseases has gained special interest through the trials carried out in the last decade. These trials seem to point at the role played by certain molecules, such as cytokines (transforming growth factor, platelet derived growth factor) and integrins, in the processes that lead to pulmonary fibrosis during the course of interstitial lung disease. They also demonstrate the important role that angiotensin II plays in increasing the secretion of transforming growth factor by several cells. The above-mentioned studies allow new therapeutic approaches to be considered which will possibly improve the serious prognosis of such diseases once they have reached the last stage of their course: pulmonary fibrosis. PMID- 14563276 TI - [The golden helix]. PMID- 14563277 TI - [Secondary prevention of acute myocardial infarction: should oral anticoagulants and platelet antiaggregants be administered on a combined basis?]. PMID- 14563278 TI - [Cardiac toxicity by carboplatin]. PMID- 14563279 TI - [Basal cell carcinoma in tanning salons users: risks of the use of ultraviolet devices]. PMID- 14563280 TI - [Diagnosis and pharmacological treatment of attention-deficit/hyperactivity disorder]. PMID- 14563281 TI - [Aseptic meningitis and Sweets syndrome]. PMID- 14563283 TI - [Interaction between methadone and antiretrovirals (stavudine, indinavir, ritonavir, nevirapine)]. PMID- 14563284 TI - [Stem cells to regenerate cardiac tissue in heart failure]. AB - Myocardial regeneration is one of the most promising therapeutic strategies for heart failure patients. Many experimental studies have demonstrated that different types of stem cell can differentiate into myocardial cells and tissues necessary for regeneration of the damaged myocardium, while studies in experimental animals suggest that muscle (myoblast), bone marrow (mesenchymal, endothelial or hematopoietic progenitors) and even heart cells can help to improve heart contractility in vivo. These findings have led several groups to undertake studies in patients with myocardial infarction. However, the use of cellular therapy in clinical trials is not without controversy, mainly related with the need for better knowledge before these therapeutic strategies are used in clinical practice. Although significant enhancement of our knowledge of the processes involved is fundamental, we do not consider it unreasonable to initiate clinical trials in which specific questions are posed, whose answers will allow us to make further progress. PMID- 14563285 TI - [Clinical practice guidelines for hypertension 2003. Do they clarify or confuse?]. AB - In May 2003 the almost simultaneous publication of the Update on Clinical Practice Guidelines by The Spanish Society of Cardiology, an initial document of the VII Report of Joint National Committee (JNC), and the European Society of Cardiology and European Society of Hypertension Guidelines in another jointly issued document, created controversy. The points of disagreement basically concerned the classification of hypertension, the choice of treatment strategy on the basis of individual cardiovascular risk, and the choice of initial treatment. A detailed analysis of the three documents, however, reveals more points of concurrence than of actual disagreement. A category between normal blood pressure and established hypertension, classified as prehypertension in the VII JNC Report and as normal-high BP in the Spanish and European Society of Cardiology Guidelines, includes a population at high risk for developing hypertension and in which lifestyle modifications are needed. In some specific clinical situations, basically in vascular high-risk patients, there is broad consensus on the definition of therapeutic groups based on scientific evidence from large clinical trials. In patients not included in a group with a specific indication, any drug included in the 5 main therapeutic groups can be used as the first step in treatment; in this group of patients thiazide diuretics play a preponderant role. The main objective of hypertension treatment is to obtain maximal reduction in overall cardiovascular risk, which requires correction of all associated risk factors and appropriate treatment for target organs likely to be affected. PMID- 14563286 TI - [Influenza vaccination: a new indication of an old therapy?]. PMID- 14563287 TI - [Neovascularization in atherosclerotic lesions: homeostatic response or mechanism of progression of the disease?]. PMID- 14563288 TI - [Flu vaccination in patients with acute coronary syndromes: treatment benefit in prespecified subgroups]. AB - BACKGROUND: The first prospective clinical Flu Vaccination in Acute Coronary Syndromes (FLUVACS) Trial has provided some evidence that flu vaccination together with standard therapy may be useful during the winter season to reduce the risk of death and major cardiac events in patients with acute myocardial infarction. PATIENTS AND METHOD: Information available in the FLUVACS database was analyzed to evaluate the efficacy of flu vaccination in different subgroups. Logistic regression was used to identify features related with better therapeutic results. RESULTS: Flu vaccination was effective in reducing the incidence of the composite endpoint (death, nonfatal myocardial reinfarction or recurrent angina prompting urgent revascularization) in most subgroups at 6 months after inclusion. The regression model showed a greater benefit of flu vaccination in patients with no ST-segment elevation or older than 65 years, nonsmokers and patients with a TIMI risk score higher than 6. CONCLUSIONS: Our data suggest that vaccination for secondary prevention of flu during the acute phase of myocardial infarction may be effective in a broad range of patients with acute coronary artery disease, regardless of their initial clinical risk. PMID- 14563289 TI - [Emergency room risk stratification of patients with chest pain without ST segment elevation]. AB - OBJECTIVES: To investigate the prognostic factors in patients who come to the emergency room with chest pain but without ST segment elevation. PATIENTS AND METHOD: 743 consecutive patients were evaluated by recording clinical history, electrocardiogram and troponin I determination, and early (<24 h) exercise testing was done for the low-risk subgroup of patients (n=203). All patients were followed during 3 months for major events (acute myocardial infarction or death). RESULTS: Major events occurred in 71 patients (9.6%). Multivariate analysis (C statistic=0.79; 95% CI 0.73-0.84; p=0.0001) identified the following predictors: age > or =72 years (OR=1.7; 95% CI, 1.0-2.9; p=0.05), insulin-dependent diabetes mellitus (OR=2.9; 95% CI, 1.5-5.4; p=0.001), previous ischemic heart disease (OR=1.9; 95% CI, 1.1-3.2; p=0.02), ST depression (OR=2.1; 95% CI, 1.2-3.8; p=0.01) and troponin I elevation (OR=2.9; 95% CI, 1.5-5.3; p=0.001). These five predictors were used to construct a risk score based on their odds ratios, which allowed event rate stratification by quartiles of the score: 0-2 points (1.6% events), 3-4 points (8.1% events), 5-7 points (11.9% events) and > or =8 points (26.2% events); p=0.0001. No patient with negative findings in the early exercise testing had major events. CONCLUSIONS: In patients with chest pain, the combination of clinical, electrocardiographic and biochemical data available on admission to the emergency service allows rapid prognostic stratification. Early exercise testing is advisable for the final stratification of low risk patients. PMID- 14563291 TI - [A prospective protocol increases oral anticoagulant prescription in patients with chronic, nonvalvular atrial fibrillation]. AB - INTRODUCTION AND OBJECTIVES: Observational studies have shown that oral anticoagulants (OAC) prescription is suboptimal in patients with nonvalvular atrial fibrillation (NVAF). Our objective was to evaluate the usefulness of a prospective protocol for increasing OAC usage in these patients. PATIENTS AND METHOD: From 1 February 2000 until 31 October 2002 we enrolled all patients with chronic NVAF seen in two outpatient cardiology clinics, excluding candidates for cardioversion. Each patient was studied to identify cardioembolic risk factors (CERF) and contraindications for OAC. Anticoagulation was suggested to all patients with > or =2 CERF and without contraindications for OAC. The decision to prescribe OAC was made by the physician in charge when there was only one CERF. RESULTS: 721 patients fulfilled the inclusion criteria. Mean age was 73 +/- 8 years; 44% were men. In most cases NVAF was related with hypertension (57%), followed by no structural heart disease (25%) or ischemic heart disease (9%). The most frequent CERFs were hypertension (66%), age > or =75 years (45%) and diabetes (24%). A total of 663 patients had > or =1 CERF (92%), and 125 (19%) of these presented at least one contraindication for OAC. Of the 538 remaining patients (90%), 485 (67% of the whole series) were treated with anticoagulation. Of the patients with > or =2 CERF and without contraindications for OAC, 95% were treated. CONCLUSIONS: A prospective protocol for use in the outpatient cardiology clinic allows to prescribe OAC in a large percentage of patients with NVAF. PMID- 14563292 TI - [Neovascularization in human coronary arteries with lesions of different severity]. AB - INTRODUCTION AND OBJECTIVES: Endothelial function can be modulated by growth factors produced by activated smooth muscle cells, inflammatory cells and plasma products that infiltrate the lesion. The aim of this study was to quantify neovessels in human coronary arteries with atherosclerotic lesions of different severity and analyze their relationship with inflammatory cell and plasma product infiltrates. PATIENTS AND METHOD: We studied 60 coronary arteries from patients who underwent heart transplant. Cellular markers (smooth muscle cell, monocyte/macrophage), the presence thrombin/prothrombin and expression of vascular endothelial growth factor (VEGF) were analyzed and quantified by conventional histology, immunohistochemistry and image analysis techniques. RESULTS: Neovessels were detected in advanced lesions, and a positive correlation was observed with the degree of vessel remodeling, monocyte/macrophage infiltration and lipid deposition. Smooth muscle cells were the main producers of VEGF in both the intima and media layers of advanced lesions. In these lesions thrombin/prothrombin-positive areas colocalized with activated smooth muscle cells. CONCLUSIONS: The presence of neovessels in coronary arteries correlated with inflammatory cell infiltration, lipid deposition and thrombin/prothrombin content. VEGF expression was mainly associated with smooth muscle cells, indicating a key role of these cells in the modulation of endothelial cell function. PMID- 14563290 TI - [Lesions produced by radiofrequency ablation of the cavotricuspid isthmus in an experimental model]. AB - INTRODUCTION AND OBJECTIVES: Experimental studies have shown that deeper and wider lesions (up to 10 mm long or deep) can be safely created using an 8 mm or irrigated tip catheter for ablation to treat atrial flutter. However, potential damage to the tricuspid valve or inferior cava vein has not been systematically evaluated. PATIENTS AND METHOD: The cavotricuspid isthmus was ablated in 26 pigs (body weight 26-52 kg), with a total of 187 radiofrequency pulses. Standard 4 mm, 8 mm and irrigated tip catheters were used at random. For each ablation, energy, impedance and temperature were recorded continuously. RESULTS: The lesions were larger with irrigated tip and 8-mm catheters than with standard ones. In 7 animals (1 with an irrigated tip, 4 with an 8-mm, and 2 with a standard tip) the tricuspid valve was damaged. The tricuspid valve was severely damaged in 3 pigs and lesions were moderate in 4. In animals with tricuspid valve lesions, maximal energy was higher (59 +/- 27 vs. 51 +/- 24 W; p=0,03) and higher temperatures were reached (63 +/- 4 vs. 55 +/- 11 degrees C; p<0.001). Low energy pulses measured before ablation were also more intense in animals in which damage was produced (0.55 +/- 0.24 vs. 0.35 +/- 0.29; p=0.001), indicating greater contact pressure. CONCLUSIONS: The tricuspid valve may be severely damaged during the ablation of the cavotricuspid isthmus for atrial flutter: damage was seen most often with high energy pulses and with 8-mm catheters, but can also occur with usual energy levels and standard catheters. To minimize damage this technique should not be used from the inside of the right ventricle just above the tricuspid valve. PMID- 14563293 TI - [Prognostic role of systemic hypertension and diabetes mellitus in patients with unstable angina undergoing coronary stenting]. AB - BACKGROUND AND OBJECTIVES: The adverse effects of systemic hypertension and diabetes mellitus in coronary patients are well known, although their long-term prognostic influence on patients with unstable angina (UA) undergoing percutaneous coronary intervention (PCI) with coronary stenting is uncertain. The aim of this study was to determine the influence of these pathologies in this population at 3-year follow-up. PATIENTS AND METHOD: We studied 279 consecutive patients with UA who underwent coronary stenting. 129 (46.2%) of them had hypertension and 60 (24.7%) had diabetes. Clinical follow-up was obtained in 92.14% after 3 years. RESULTS: Although the need for new PCI at the target lesion was higher for patients with hypertension and diabetes (12.1 vs 8.4%; p=0.31, and 14.5 vs 8.6%; p=0.16, respectively), the differences were not significant with respect to the control groups. Multivariate analysis showed hypertension (OR=4.71; CI 95%, 1.01-42.2; p=0.04) and ejection fraction (OR=0.95; CI 95%, 0.91 0.99; p=0.03) to be predictors of mortality, and diabetes to be a predictor of myocardial infarction and infarction resulting in death (OR=3.01; CI 95%, 1.13 8.02; p=0.02, and OR=2.68; CI 95%, 1.03-6.95; p=0.04, respectively). CONCLUSIONS: Hypertension was the only independent long-term predictor of mortality in our series of patients with UA who underwent coronary stenting. Diabetes was the only predictor of myocardial infarction or for the combined event of infarction and death. Risk of myocardial infarction was threefold as high in this diabetic patient population, and was the main cause of mortality. PMID- 14563294 TI - [Predictors of myocardial contractile reserve in patients with nonischemic dilated cardiomyopathy. An echo-stress dobutamine study]. AB - INTRODUCTION AND OBJECTIVES: Myocardial contractile reserve studies with low-dose dobutamine echocardiography have been shown to be useful to assess functional myocardial status. However, the variables associated with contractile reserve after inotropic stimulation are not well known. PATIENTS AND METHOD: We studied 50 patients (35 men, mean age 56.4 +/- 9.5 years) with nonischemic dilated cardiomyopathy (NIDC), LVEF 28.7% +/- 8.5% and wall motion score index (WMSI) 2.42 +/- 0.34 with low-dose dobutamine echocardiography. Left ventricular contractile reserve was assessed by a differential parameter defined as the difference between rest and stress WMSI (DeltaWMSI). RESULTS: After dobutamine infusion the WMSI was 1.95 +/- 0.58; from this value we calculated a DeltaWMSI of 0.45 +/- 0.39. None of the clinical variables showed a relationship with the presence of contractile reserve. In contrast, the following echocardiographic parameters correlated with DeltaWMSI: end-diastolic (p=0.05) and end-systolic (p=0.02) diameters, end-systolic volume index (p=0.01) and LVEF (p=0.002). In the multivariate analysis, only end-diastolic diameter was an independent predictor of contractile reserve (hazard ratio=0.852; 95% CI, 0.735-0.987; p=0.03). CONCLUSIONS: Ventricular diameters, end-systolic volume index and LVEF are related with improvements in myocardial contractility after dobutamine infusion, although only end-diastolic diameter was an independent predictor of contractile reserve. Thus, this parameter should receive particular attention in evaluations of the functional status of the myocardium in patients with NIDC. PMID- 14563295 TI - [Role of coronary risk factors in blood thrombogenicity and acute coronary syndromes]. AB - Recent advances in basic science have linked some systemic risk factors to endothelial dysfunction which gives rise to atherosclerotic disease and triggers the progression of thrombotic complications. Superficial erosion of the stenotic plaque can be observed in one-third of acute coronary syndromes (ACS). In these cases the presence of classic risk factors such as diabetes mellitus, hypercholesterolemia and smoking favor a state of "vulnerable blood" or high risk. Increased thrombogenicity can exacerbate thrombus formation and is able to trigger an ACS. The vessel endothelium regulates contractile, mitogenic and thrombotic activities of the vessel wall. Risk factors impair both homeostasis and hemostasis of the vessel wall and promote inflammatory signals. Platelet and monocyte activation favors the expression of tissue factor (TF), thus triggering the coagulation cascade with thrombin generation and clot formation. Increased blood thrombogenicity linked to classic risk factors may be associated with circulating TF levels which are much higher than those observed in healthy subjects without risk factors. These observations not only emphasize the usefulness of aggressive management of risk factors but open a new avenue for future studies to devise therapeutic strategies to treat ACS by inhibiting TF expression. PMID- 14563296 TI - [Rational use of noninvasive cardiac stress testing in the diagnosis of coronary artery disease]. AB - A variety of noninvasive tests are available to clinicians for the evaluation of patients in whom ischemic heart disease is suspected because of chest pain, clinical antecedents, or a combination of the two. Although all tests in general help to varying degrees to refine (by inclusion or exclusion) the diagnosis in a given patient, there are undoubtedly important differences between tests regarding their scope and diagnostic accuracy in general, and with respect to certain groups of patients in particular. Because of this, and in view of the obvious economic implications, the topic merits critical review before the information obtained from these tests is used in patient management. This review is not intended to cover all features that argue for or against all currently available noninvasive tests for ischemic heart disease, but to place into perspective the importance of the clinical assessment of the patient in the light of the results of testing, and to obtain a more rational idea of their usefulness. Despite the risk of excluding certain material of interest, excellent techniques such as magnetic resonance imaging will not be covered in the review, only because they have not yet been included in meta-analyses. Emphasis on the Bayesian rationale or paradigm, together with discussion of recent meta-analyses, offers a balanced perspective of the use and possible misuse of these diagnostic tests, and of their clinical and economic implications. PMID- 14563297 TI - [Percutaneous mitral valvuloplasty guided by three-dimensional echocardiography]. PMID- 14563298 TI - [Spontaneous coronary artery dissection. An infrequent cause of acute coronary syndromes]. AB - Spontaneous coronary artery dissection is a rare condition that may produce severe myocardial ischemia. The growth of indications for cardiac catheterization have led to an increment in the number of cases identified in patients with acute coronary syndromes. Because the therapeutic approach and prognosis are uncertain, doubts often arise regarding the optimal management of these patients. We describe here the clinical and angiographic characteristics of 7 patients with spontaneous coronary artery dissection, as well as treatment and follow-up. PMID- 14563299 TI - [Direct detection of malignant mutations in patients with hypertrophic cardiomyopathy]. AB - We determined the prevalence of mutations considered malignant in the genes for beta-myosin heavy chain (MYH7, 11 mutations) and troponin T (TNNT2, 5 mutations) in 30 patients with hypertrophic cardiomyopathy aged 18 to 60 years, 83% of whom had familial antecedents of hypertrophic myocardiopathy or sudden death. Mutations were identified with polymerase chain reaction followed by restriction enzyme digestion and agarose gel electrophoresis. Direct analysis identified 16 mutations in 2 of the 30 patients (7%): one women diagnosed at the age of 25 years as carrying the MYH7453cysteine mutation, and a 60-year-old women with the TNNT2278 cysteine mutation. These cases illustrate the considerable clinical heterogeneity that characterizes carriers of these mutations. Clinical manifestations can range from severe hypertrophy or early sudden death to the absence of symptoms up to advanced age. PMID- 14563300 TI - [Distal intercoronary communication. A case report and medical literature review]. AB - Direct connections between normal distal coronary arteries are rarely detected with angiography. In a 67-year-old man a diagnosis of intercoronary arterial continuity was established after ruling out collateral circulation secondary to coronary obstruction. Published reports of similar cases are reviewed. PMID- 14563303 TI - [Clinical episode of suggestive left main coronary artery spasm after gingival anaesthetic infiltration]. PMID- 14563304 TI - [Design and conclusions of the ALLHAT study]. PMID- 14563306 TI - Collaborative study for establishment of the European Pharmacopoeia BRP batch 1 for diphtheria toxin. AB - A stable liquid candidate Biological Reference Preparation (BRP) for diphtheria toxin was prepared in peptone buffer (nominal content of diphtheria toxin: 1 Lf/ml, 0.4 micro g/ml), filled in ampoules (filling volume: 1 ml) and characterised in a collaborative study. The toxin is to be used in the test "Absence of toxin and irreversibility of toxoid" as described in the current European Pharmacopoeia (Ph. Eur.) monograph Diphtheria Vaccine (Adsorbed) (2002:0443). Eleven laboratories assessed the specific activity of the preparation by in vivo and in vitro assays. The material is assumed to have satisfactory stability with a calculated predicted loss of activity of <1% per year at 4-8 degrees C. From the collaborative study, the specific activity was calculated as 77.6 (45-113) LD( 50)/ml (lethal challenge) and >75 000 Lr/Lf (intradermal challenge). The candidate BRP was successfully used in nine laboratories and confirmed suitable for use in the Vero cell test for "Absence of toxin and irreversibility of toxoid" as described in the Ph. Eur. monograph 2002:0443; i.e., concentrations of 5 x 10( -5) Lf/ml and below caused cytotoxic effects in the Vero cell test. Due to its liquid nature, the stability of the material will be monitored at regular intervals and preparation of a stable freeze-dried formulation will be considered for long-term use. Additional studies will be performed to confirm suitability of this BRP for other applications. The candidate BRP was adopted as the Ph. Eur. reference material for Diphtheria Toxin Batch 1 by the Ph. Eur. Commission at its session in March 2003. PMID- 14563307 TI - Establishment of European Pharmacopoeia BRP batch 2 for inactivated poliomyelitis vaccine for in vitro D antigen assay. AB - A collaborative study was initiated by the European Directorate for the Quality of Medicines (EDQM) to assign a potency value for the candidate Ph Eur BRP batch 2 against the 2nd International Standard (IS) in order to replace the dwindling stocks of Ph Eur BRP batch 1. The candidate material is a concentrated trivalent bulk (Type 1 (Mahoney), Type 2 (MEF1) and Type 3 (SAUKETT)) from a commercially available IPV vaccine. Nine laboratories participated in the collaborative study. Eight laboratories reported results. Participants performed in-house ELISA assays on the candidate BRP, the 2nd International Standard (IS) and the current BRP (BRP batch 1). An additional sample was included to acquire information on the correlation between the in vitro and in vivo assays based on comparison with a previous study. Results of that comparison are included as an annex. Potency estimates were satisfactory in terms of repeatability and reproducibility, however the estimates for the 2nd IS were significantly lower than those for Ph Eur BRP batch 1. These two reference standards are derived from the same material and were originally assigned the same potency value after a joint study run by EDQM and the WHO in 1994. A reconciliation study was therefore designed to determine if the IS stored at NIBSC and the IS which had been sent from NIBSC to EDQM for use in the initial study were equivalent. 3 of the laboratories from the initial study participated. Results revealed no significant difference between the 2nd IS stocks stored in the two different locations at NIBSC nor between BRP batch 1 and the standards stored at NIBSC for types 1 and 2. For type 3 the 2nd IS standards stored at NIBSC are 13 % less potent than the Ph Eur BRP batch 1. The 2nd IS which had been shipped from NIBSC to EDQM was significantly less potent than BRP batch 1 and the 2nd ISs stored at NIBSC for all three types, confirming the observation of the initial study. Possible explanations for this apparent loss of potency of the 2nd IS used in the study are under investigation. Since Ph Eur BRP batch 1 and the 2nd IS in stock at NIBSC appear no more different than when their original potency assignment was made at their establishment, and since the 2nd IS standard used in the initial part of this study was compromised, a consensus potency value for the candidate BRP was determined using Ph Eur BRP batch 1 as the reference standard. The candidate material was therefore assigned a potency of 320-67-282 D Antigen units/ml (IU) for types 1, 2 and 3 respectively. A stability monitoring program will be initiated. The candidate material was adopted by the European Pharmacopoeia Commission at its session in March 2003 as European Pharmacopoeia IPV vaccine BRP batch 2 for D Ag in vitro assay. PMID- 14563308 TI - Feasibility study to evaluate the correlation between results of a candidate in vitro assay and established in vivo assays for potency determination of Newcastle disease vaccines. AB - A Newcastle disease virus antigen quantification assay has been developed at CIDC Lelystad as a candidate in vitro potency test for inactivated Newcastle disease vaccines. In studies performed at CIDC-Lelystad, a high correlation was demonstrated between the results of this candidate in vitro potency assay and the results of the serological potency assay (European Pharmacopoeia monograph 0870; test A). Furthermore, a high correlation between the serological data (Haemagglutination Inhibition-antibody titres) and clinical protection after challenge was demonstrated. The aim of the feasibility study was to confirm the correlation between the results obtained using the candidate in vitro potency assay and the results from both the in vivo potency assays currently prescribed in Ph Eur monograph 0870, in different laboratories and to determine whether a large-scale validation study of the in vitro method should ensue. In the feasibility study three Official Medicines Control Laboratories tested the potency of 5 different inactivated Newcastle disease vaccines and one experimental vaccine, using both of the in vivo methods described in the European Pharmacopoeia and the candidate in vitro method. The 6 vaccine batches represented a quantitative range of Newcastle disease virus antigen content and were produced by different manufacturers. Statistical evaluation of all results indicated that a satisfactory correlation was found in all laboratories between the two types of in vivo tests currently in place, and the candidate in vitro test. An excellent reproducibility of the proposed in vitro method was observed with respect to the ranking of the vaccines included in this study. It is concluded that the results of this feasibility study indicate that a large-scale collaborative study can be organised to validate the in vitro method and the suitability of the reference preparation. PMID- 14563309 TI - Glimpses at TCR trans-species crossreactivity. PMID- 14563310 TI - A novel means of favorably tipping the balance between cytopathic and regulatory T cells. PMID- 14563311 TI - It's a good year for Blimp-1 (and plasma cells). PMID- 14563312 TI - Stress management: MHC class I and class I-like molecules as reporters of cellular stress. AB - The evolutionarily ancient intracellular stress response protects cells from the effects of external and internal forces which perturb cellular metabolism. Members of the major histocompatibility complex (MHC) class I-like superfamily act as cell surface indicators of the intracellular stress response. Cellular immunity employs these indicators as a cue for elimination of damaged, infected, and malignant cells, promoting the health of the individual and the evolutionary success of the species. PMID- 14563313 TI - Induction of somatic hypermutation is associated with modifications in immunoglobulin variable region chromatin. AB - Somatic hypermutation (SHM) requires selective targeting of the mutational machinery to the variable region of the immunoglobulin heavy chain gene. The induction of SHM in the BL2 cell line upon costimulation is associated with hyperacetylation of the chromatin at the variable region but not at the constant region. The V region-restricted histone hyperacetylation resulting from costimulation occurs independent of AID expression and mutation. Interestingly, costimulation in the presence of Trichostatin A causes hyperacetylation of histones associated with the constant region and extends mutations to the constant region. Under this condition, promoter proximal mutations are observed in the variable region as well. The overexpression of AID results in a similar deregulation of mutational targeting. Our results indicate that the stimulation of SHM in BL2 cells activates two independent pathways resulting in histone modifications that permit induced levels of AID to selectively target the variable region for mutation. PMID- 14563314 TI - Intracellular location and cell context-dependent function of protein kinase D. AB - Protein kinase D (PKD) is an antigen receptor-activated serine kinase localized at either the plasma membrane or the cytosol of lymphocytes. To probe PKD function at these different locations, transgenesis was used to target active PKD either to the membrane or cytosol of pre-T cells. In recombinase gene null pre-T cells, membrane and cytosolic active PKD both induced differentiation reminiscent of beta selection: downregulation of CD25 and upregulation of CD2 and CD5. Active PKDs also induced pre-T cell proliferation, although this response was not universal to all thymocyte subsets. There were two striking differences between the actions of the differentially localized PKDs. Membrane but not cytosolic PKD could induce expression of CD8 and CD4 in recombinase null mice; cytosolic but not membrane PKD suppressed Vbeta to DJbeta rearrangements of the TCRbeta chain locus in wild-type T cells. PKD function is thus determined by its intracellular location and cell context. PMID- 14563315 TI - Favorably tipping the balance between cytopathic and regulatory T cells to create transplantation tolerance. AB - Therapeutic application of broadly reactive anti-T cell antibodies can lead not only to potent immunosuppression but also to profound and long-lived T cell depletion. We reasoned that a strategy that almost exclusively targets activated cytopathic donor reactive T cells and spares immunoregulatory networks might prove to be an exceptionally potent and highly selective means of producing long term engraftment and tolerance. Herein we show that the combined administration of rapamycin and agonist IL-2- and antagonist IL-15-related cytolytic fusion proteins provides for long-term engraftment/tolerance in exceptionally stringent allotransplant models by (1) limiting the early expansion of activated T cells, (2) preserving and even exaggerating their subsequent apoptotic clearance, and (3) further amplifying the depletion of these activated T cells by antibody dependent mechanisms, while (4) preserving CD4+CD25+ T cell-dependent immunoregulatory networks. PMID- 14563317 TI - The endoglin(positive) sca-1(positive) rhodamine(low) phenotype defines a near homogeneous population of long-term repopulating hematopoietic stem cells. AB - Endoglin, an ancillary TGF-beta receptor, is differentially expressed in long term repopulating hematopoietic stem cells (LTR-HSC). Here, we describe simple and highly efficient purification schemes for mouse bone marrow LTR-HSCs using Endoglin as a marker. The Endoglin positive and Sca-1 positive (Endo(Pos) Sca 1(Pos)) population, which contains about 36% of "Side Population" (SP) cells, is highly enriched for LTR-HSCs. In long-term competitive reconstitution assays, 100 such cells reconstituted all lethally irradiated recipients. Interestingly, the Endo(Pos) Sca-1(Pos) population contains comparable LTR-HSC activity in both SP and non-SP fractions, indicating that many HSCs are not captured by the SP phenotype. Furthermore, LTR-HSCs are exclusively found in the Endo(Pos) Sca 1(Pos) Lin(Neg/Low) (lineage negative/low), but not in the Endo(Neg) Sca-1(Pos) Lin(Neg/Low) population, suggesting that the Endo(Pos) population may contain all LTR-HSCs in mouse bone marrow. Finally, we demonstrated that the Endo(Pos) Sca 1(Pos) Rh(Low) (Rhodamine-123 low) phenotype, without using CD34, c-Kit, or Lineage markers, defines a nearly homogenous population of LTR-HSCs. PMID- 14563316 TI - The Smu tandem repeat region is critical for Ig isotype switching in the absence of Msh2. AB - Deficiencies of the Msh2 protein or the Smu tandem repeat (SmuTR) sequences each reduce isotype switching in mice by about 2- to 3-fold. We find that switching in mice deficient for both Msh2 and SmuTR is nearly ablated. We propose that the SmuTR provides closely spaced cleavage sites that can undergo switch recombination independent of Msh2, whereas cleavages in sequences flanking the SmuTR require Msh2 processing to allow recombinational joining. We also find that changes in Smu sequences alter the focus of switch junctions within Sgamma sequences, indicating that sequences of switch regions act together in the choice of switch recombination junctions. These findings help to explain the conservation of tandemly repeated switch regions associated with heavy chain constant genes in species capable of switching. PMID- 14563318 TI - Production of ribosome components in effector CD4+ T cells is accelerated by TCR stimulation and coordinated by ERK-MAPK. AB - Effector CD4+ T cells rapidly activate high-level cytokine expression following TCR stimulation. Consistent with accelerated protein production in these cells, global mRNA profiles revealed that, after cytokines, the most impressive cluster of activated genes encode rRNA-maturation factors. Activation of these genes was ERK-MAPK dependent, accompanied by increased rRNA transcription and faster maturation kinetics, and much greater in effector CD4+ T cells than in naive cells. Ribosomal protein subunit (RPS) synthesis was also ERK-MAPK dependent and increased to match rRNA production, but without evident increase in RPS mRNA. Instead, stimulation promoted polysome loading of RPS mRNA via cis-acting, 5' terminal oligopyrimidines. These results demonstrate how, in response to extracellular signals, effector CD4+ T cells coordinately increase multiple ribosomal components to accommodate burgeoning cytokine production. PMID- 14563319 TI - Delta1-Notch3 interactions bias the functional differentiation of activated CD4+ T cells. AB - Following activation by antigen, naive CD4+ T helper precursor cells execute distinct genetic programs that result in their differentiation toward the type 1 or type 2 helper T cell (Th1 or Th2) phenotype. Although the differentiation and function of these Th subsets has been well studied, little is known about the contribution to these differentiation events of cell surface receptors other than those for soluble cytokines, such as IL-12 or IL-4. Here, we provide direct evidence that the Delta1 interaction with Notch3 on CD4+ T cells transduces signals, promoting development toward the Th1 phenotype. The positive role of Notch signaling in effector cell differentiation was dose dependent, with high levels of stimulation resulting in reduced T cell activation. Our data revealed a clear contribution of Notch pathways to Th1 versus Th2 fate decisions, while also providing insight into another mechanism for inhibition of CD4+ T cell activation. PMID- 14563320 TI - Natural killer cells activated by MHC class I(low) targets prime dendritic cells to induce protective CD8 T cell responses. AB - Conserved molecular patterns derived from pathogenic microorganisms prime antigen presenting dendritic cells (DC) to induce adaptive T cell responses. In contrast, virus-infected or tumor cells that express low levels of major histocompatibility complex (MHC) class I activate natural killer (NK) cells for direct killing. It is unknown whether NK cell recognition of MHC class I(low) targets can also induce adaptive T cell responses. Here, we show that MHC class I(low) targets initiate a cascade of immune responses, starting with the immediate activation of NK cells. The activated NK cells then prime DC to produce IL-12 and to induce highly protective CD8 T cell memory responses. Therefore, sensing of MHC class I(low) targets by NK cells can link innate and adaptive immunity to induce protective T cell responses and may alarm the immune system during early infection with noncytopathic viruses. PMID- 14563321 TI - NAD-induced T cell death: ADP-ribosylation of cell surface proteins by ART2 activates the cytolytic P2X7 purinoceptor. AB - T cells express a toxin-related ADP-ribosylating ectoenzyme, ART2. Exposure of mature T cells to NAD, the substrate for ADP-ribosylation, induces cell death. ART2-catalyzed ADP-ribosylation activates the cytolytic P2X7 purinoceptor, causing calcium flux, pore formation, phosphatidylserine exposure, shedding of CD62L, cell shrinkage, and propidium iodide uptake. Interestingly, much lower NAD than ATP concentrations are required to activate P2X7. NAD-induced cell death (NICD) operates with endogenous sources of NAD released upon cell lysis. These findings identify P2X7 as a key effector of NICD and demonstrate that P2X7 can be activated by an endogenous ligand other than ATP. Our results delineate an alternative mechanism for inducing T cell death and set an interesting precedent for immunoregulation via crosstalk between NAD-dependent ADP-ribosyltransferases and purinoceptors. PMID- 14563322 TI - Chemokines acting via CXCR2 and CXCR4 control the release of neutrophils from the bone marrow and their return following senescence. AB - In this study we provide evidence that the SDF-1alpha/CXCR4 chemokine axis is involved in both the retention of neutrophils within the bone marrow and the homing of senescent neutrophils back to the bone marrow. We show that the functional responses of freshly isolated human and murine neutrophils to CXCR2 chemokines are significantly attenuated by SDF-1alpha, acting via CXCR4. As a consequence, the mobilization of neutrophils from the bone marrow in vivo by the CXCR2-chemokine, KC, was dramatically enhanced by blocking the effects of endogenous SDF-1alpha using a specific CXCR4 antagonist. As neutrophils age, they upregulate expression of CXCR4 and acquire the ability to migrate toward SDF 1alpha. We show here that these senescent CXCR4(high) neutrophils preferentially home to the bone marrow in vivo in a CXCR4-dependent manner, suggesting a previously undefined mechanism for the clearance of senescent neutrophils from the circulation. PMID- 14563323 TI - A correlation between TCR Valpha docking on MHC and CD8 dependence: implications for T cell selection. AB - T cell receptors (TCR) adopt a similar orientation when binding with major histocompatibility complex (MHC) molecules, yet the biological mechanism that generates this similar TCR orientation remains obscure. We show here the cocrystallographic structure of a mouse TCR bound to a human MHC molecule not seen by the TCR during thymic development. The orientation of this xenoreactive murine TCR atop human MHC deviates from the typical orientation more than any previously determined TCR/MHC structure. This unique orientation is solely due to the placement of the TCR Valpha domain on the MHC. In light of new information provided by this structure, we have reanalyzed the existing TCR/MHC cocrystal structures and discovered unique features of TCR Valpha domain position on class I MHC that correlate with CD8 dependence. Finally, we propose that the orientation seen in TCR recognition of MHC is a consequence of selection during T cell development. PMID- 14563324 TI - Blimp-1 is required for the formation of immunoglobulin secreting plasma cells and pre-plasma memory B cells. AB - Blimp-1 is a transcriptional repressor able to drive the terminal differentiation of B cells into Ig-secreting plasma cells. We have created mice with a B cell specific deletion of prdm1, the gene encoding Blimp-1. B cell development and the number of B cells responding to antigen appear to be normal in these mice. However, in response to either TD or TI antigen, serum Ig, short-lived plasma cells, post-GC plasma cells, and plasma cells in a memory response are virtually absent, demonstrating that Blimp-1 is required for plasmacytic differentiation and Ig secretion. In the absence of Blimp-1, CD79b(+)B220(-) pre-plasma memory B cell development is also defective, providing evidence that this subset is an intermediate in plasma cell development. B cells lacking Blimp-1 cannot secrete Ig or induce muS mRNA when stimulated ex vivo. Furthermore, although prdm1-/- B cells fail to induce XBP-1, XBP-1 cannot rescue plasmacytic differentiation without Blimp-1. PMID- 14563325 TI - Bam32 links the B cell receptor to ERK and JNK and mediates B cell proliferation but not survival. AB - Bam32 is an adaptor protein recruited to the plasma membrane upon B cell receptor (BCR) crosslinking in a phosphoinositol 3-kinase (PI3K)-dependent manner; however, its physiologic function is unclear. To determine its physiologic function, we produced Bam32-deficient mice. Bam32(-/-) B cells develop normally but have impaired T-independent antibody responses in vivo and diminished responses to BCR crosslinking in vitro. Biochemical analysis revealed that Bam32 acts in a novel pathway leading from the BCR to MAPK/ERK Kinases (MEK1/2), MAPK/ERK Kinase Kinase-1 (MEKK1), extracellular signal-regulated kinase (ERK), and c-jun NH2-terminal kinase (JNK), but not p38 mitogen-activated protein kinase (p38). This pathway appears to be initiated by hematopoietic progenitor kinase-1 (HPK1), which interacts directly with Bam32, and differs from all previously characterized BCR signaling pathways in that it is required for normal BCR mediated proliferation but not for B cell survival. PMID- 14563327 TI - Maxillofacial fractures resulting from falls. AB - OBJECTIVE: This study was performed to clarify the characteristics of facial fractures caused by falls with a particular focus on aetiology. PATIENTS: Of 505 patients with facial fractures treated in the Department of Oral and Maxillofacial/Craniofacial Surgery, Heidelberg University Hospital from January 1997 to May 2001, 129 patients injured by falls were analysed. STUDY DESIGN: Aetiological analysis was performed according to falling patterns. RESULTS: The distribution of age showed two peaks in the 4th and 8th decades, and the male to female ratio was 1.1:1, unlike the overall facial fracture ratio of 3:1. Seventy six patients had fallen from standing height or less and 44 patients had fallen from greater heights. The former cases were more often seen in older females. More severe injuries tend to be seen in the patients who have fallen from greater heights, and in the cases associated with acute medical disorders. CONCLUSION: This retrospective study documents the higher risk of fractures in older females and the higher risk of severe injury in those patients who were unconscious. This is in agreement with recent orthopaedic studies. PMID- 14563326 TI - Laser-scan-based navigation in cranio-maxillofacial surgery. AB - BACKGROUND: In computer-assisted surgery, a correlation between a volume data set and the surgical site is required in order to localize the patient's head on the operating table. Registration markers are commonly used for this procedure. However, the marker registration is associated with high logistics, since the markers have to be placed prior to data set acquisition and have to be kept in their position until the patient enters the operating room. This study deals with a new markerless registration method in cranio-maxillofacial surgery that is based on a high-resolution laser-scan of the patient's (relaxed) skin surface. PATIENTS: 20 patients with tumours, bone malformations or foreign bodies, scheduled for computer-assisted surgery, were involved in the study. STUDY DESIGN: The clinically applied accuracy of the laser-scan-based registration was measured through additionally placed registration markers. The inherent precision of the laser-scan registration system was controlled in phantom studies. RESULTS: The clinically applied accuracy of the new laser-scan-based registration technique ranged between 0.2 and 1.8 mm with a mean deviation of 1.1mm and a standard deviation of 0.3 mm. CONCLUSION: The facial skin surface can serve as a sufficiently stable and invariable reference base in order to register patients for computer-assisted cranio-maxillofacial surgery. PMID- 14563328 TI - Comparative evaluation of two osteosynthesis methods on stability following sagittal split ramus osteotomy. AB - INTRODUCTION: The aim of this study was to employ the finite element method (FEM) to compare the stability of 2.0 mm titanium screws in a triangular configuration with that of a 2.0 mm titanium miniplate as osteosynthesis material following bilateral sagittal split osteotomy. MATERIAL: A model of the mandible was produced, consisting of 19,854 elements and 4285 nodes. The mechanical parameters of the materials investigated were taken from the literature and notified by the manufacturer. RESULTS: On condition that the materials were subjected only to their respective ultimate tensile stress, it was possible to neutralise a masticatory force of 1246 N (Newtons) with the miniplate and of 1675 N with the bicortical triangular screw configuration. The strain limit was determined by the peri-implant bone and not the osteosynthesis material. CONCLUSION: The finite element method (FEM) appears to be suitable for simulating complex mechanical stresses in the maxillofacial area, as also confirmed by the agreement between our data and those in the literature, and with clinical experience. It should enable considerable savings to be made in terms of time, material and animal experiments in the future development of osteosynthesis materials and techniques. PMID- 14563329 TI - Analysis of complications in fractures of the mandibular angle--a study with finite element computation and evaluation of data of 277 patients. AB - AIM: The purpose of this study was to compute the load on different osteosynthesis plates in a simplified model using finite element analysis, and to find out whether miniplates were sufficiently stable for application at the mandibular angle. PATIENTS: Data from 277 patients with 293 fractures of the mandibular angle have been evaluated. METHODS: A computation model using finite elements was established in order to compute mechanical stress occurring in osteosynthesis plates used for fixation of fractures of the mandibular angle. In the second part of this study, the data from all in-patients treated for fracture of the mandibular angle were evaluated retrospectively. Age and sex of the patients, cause of fracture, state of dentition, type of therapy as well as complications were noted. RESULTS: In those tests, both the 1.0 mm miniplate and the 2.3 mm module plate were sufficiently stable. The rate of major complications (requiring revisional surgery with general anaesthesia) amounted to approximately 17% in comminuted fractures, or in non-compliant patients in which primary stability with a single miniplate did not appear sufficient, so that other osteosynthesis methods were used in addition. This rate was considerably higher than that in simple mandibular fractures. Simple fractures of the mandibular angle were just treated with one miniplate following Champy's guidelines strictly. In these fractures the rate of major complications was only 2.3%. CONCLUSION: In comminuted fractures and in non-compliant patients, the use of a stronger osteosynthesis material should be considered while in all other cases application of a single 1.0 mm miniplate was regarded as sufficient for fixation using open reduction. PMID- 14563331 TI - Analysis of nasal and labial deformities in cleft lip, alveolus and palate patients by a new rating scale: preliminary report. AB - INTRODUCTION: In this study the nasal deformities in patients with cleft lip, alveolus and palate (CLAP) were analysed and the relevant role of the perinasal perioral muscular balance, and the inborn dislocation of the alar cartilages is presented. PATIENTS AND METHODS: 50 CLAP patients were analysed in whom 29 primary cheiloplasties, 12 lip revisions and 9 rhinoplasties were performed. The lip repair was done by a modification of Millard's technique, the nose by either a closed or open-sky rhinoplasty. The severity of the cleft appearance was evaluated pre- and postoperatively, according to a pre-agreed visual rating scale. There were 4 degrees of severity of the deformity preoperatively (mild, moderate, severe and very severe), and postoperatively 5 categories of outcome (excellent, very good, good, satisfactory and poor) depending on the scores obtained by summing up the points corresponding to different types of deformity. This scale is closely related to the American Cleft Palate classification of clefts. RESULTS: 17 excellent, 4 very good, 2 good, 5 satisfactory and 1 poor result were obtained in the group of primary cheiloplasty. Eight excellent, 4 very good results were obtained by the lip revisions. Seven excellent and 2 satisfactory results were obtained following rhinoplasty. CONCLUSIONS: During the primary lip repair, it is important to correct the abnormal position of ala nasi, the nasal floor and the base of the columella. When correct insertion of m. transversus nasi to the nasal spine is achieved and a good repair of m. orbicularis oris, symmetry of the alae and normal growth of lip and columella was obtained even in most severe bilateral cases. In cases of diastasis of the orbicularis and transversus nasi muscles, in combination with other soft tissue deformities or scars, a secondary musculo-periosteal revision is recommended. The defect of the soft tissue triangle of the nose is best corrected via an open rhinoplasty. PMID- 14563330 TI - Fibula microvascular free tissue reconstruction of the severely comminuted atrophic mandible fracture--case report. AB - Severely atrophic comminuted fractures of the mandible often have inadequate bone stock available to allow for adequate rigid fixation and subsequent progression to union. Grafting with rib, iliac crest or a variety of allograft materials is required in order to increase the success rate of the repair in this patient population. In this article, we report our favourable experience in using a fibula microvascular free tissue transfer for the treatment of a particularly challenging patient with a fractured atrophic mandible. Secondary implant rehabilitation completed the reconstruction. This method may represent an alternative in the treatment of fractures of the severely atrophic mandible in select individuals. PMID- 14563332 TI - Assessment of facial emotion encoding and decoding skills in children with and without oral clefts. AB - PURPOSE: Investigated the facial emotion decoding and encoding skills and perceived social acceptance in children with and without oral clefts. PATIENTS: 8 15 year-old children with surgically repaired facial clefts (n=19) and non-cleft controls (n=19). METHODS: The children viewed photographs of facial emotions expressed by other children and identified the emotion in each photograph. They were videotaped while listening to a series of brief vignettes designed to evoke facial emotions and while posing prototypic facial expressions. They also completed a measure of their perceived social acceptance. Judges, blind to experimental group and targeted emotion, viewed systematically sampled video images, then recorded their subjective judgement of the emotion expressed in each. Trained coders also scored selected images using objective measures of the degree to which specific facial movements associated with emotion-specified expressions were present. RESULTS: Analysis of variance procedures detected significantly different patterns of facial responses between the oral cleft and control groups, and within group. Pearson correlation analyses found significant relationships between specific facial movements and perceived social acceptance. CONCLUSION: Systematic assessment of facial expression in children with oral clefts can identify unique differences in their encoding of facial emotion and may lead to behavioural interventions to improve social functioning by training facial expression skills. PMID- 14563333 TI - Conservative management of otitis media in cleft palate. AB - AIMS: Eustachian tube dysfunction affects nearly all children with cleft palate but its management is controversial. Some units perform routine prophylactic grommet insertion at the time of palate repair, whilst others are more conservative, inserting grommets only when signs and symptoms of otitis media with effusion are present. This study aims to present outcome data from one cleft team practising a conservative approach. DESIGN: This is a retrospective study in which consecutive palate repairs over 10 years are analysed and compared with previously published data. The spectrum of clefting and severity (LAHSHAL), otological and speech outcomes were recorded. Patients were excluded if incomplete data was available, and if sensorineural deafness or syndromic clefting was present. RESULTS: Data is presented for 72 of 109 consecutive patients and the 37 excluded patients are discussed. Following a conservative approach to otitis media with effusion, 29% of cases required grommets. The use of grommets seemed to be more common in those with more severe clefting. Despite this, the group receiving grommets had better speech results than those who did not, although this improvement was not statistically significant. CONCLUSIONS: There is no evidence of poor overall otological outcome in this series. The data demonstrates that those receiving grommets had better results despite more severe clefting. PMID- 14563334 TI - Parry-Romberg syndrome: intracranial MRI appearances. AB - AIM: To gain further insight into the pathogenesis of Parry-Romberg syndrome, a sporadic disease of unknown aetiology characterized by progressive wasting of one side of the face. METHOD: Cranial MRI was performed in 10 patients with Parry Romberg syndrome. The central nervous system findings are correlated to clinical findings and a review of the literature. RESULTS: Three patients with a history of migraine had abnormal brain findings confined to the cerebral hemisphere ipsilateral to the facial hemiatrophy. Two patients without CNS symptoms had intracranial changes, one ipsilateral, the other both ipsilateral and contralateral to the facial hemiatrophy, on MRI. These changes consisted of either intracerebral atrophy or white matter hyperintensity. Five patients without CNS symptoms had no pathological intracranial MRI appearances. CONCLUSION: A significant number of patients with Parry-Romberg syndrome may have underlying brain involvement. These findings are consistent with previous reports. PMID- 14563335 TI - Temporomandibular region metastasis from cystosarcoma phyllodes: a case report and review of the literature. AB - A rare breast tumour (Cystosarcoma phyllodes) in a 58-year-old female metastasizing to the temporomandibular region and infra-temporal fossa, presenting as temporomandibular pain dysfunction syndrome is reported, along with a literature review. PMID- 14563337 TI - Combining glycoprotein blockers with fibrinolysis: a bold stroke? PMID- 14563338 TI - Type II secretory phospholipase A2: the emerging role of biochemical markers of plaque inflammation. PMID- 14563339 TI - Risk of intracranial haemorrhage with combined fibrinolytic and glycoprotein IIb/IIIa inhibitor therapy in acute myocardial infarction. Dichotomous response as a function of age in the GUSTO V trial. AB - BACKGROUND: Intracranial haemorrhage is an important limitation to pharmacologic reperfusion therapy for acute myocardial infarction. The combination of a glycoprotein IIb/IIIa inhibitor, half-dose plasminogen activator and reduced-dose heparin has been evaluated as an alternative to standard fibrinolytic therapy in this setting. METHODS AND RESULTS: We evaluated the relation between univariate and multivariate predictors of intracranial haemorrhage and the effect of treatment with either reteplase alone (10 U bolus twice, 30 min apart) with standard-dose heparin (5000 U bolus followed by an infusion of 1000 Uh(-1)for patients > or =80 kg and 800 Uh(-1)for those <80 kg) or combination therapy with abciximab (0.25mg/kg bolus and 0.125 microg/kg/min for 12h) and half-dose reteplase (two boluses of 5U 30 min apart) with reduced-dose heparin (60 Ukg( 1)bolus, maximum 5000 U, followed by an infusion of 7 Ukg(-1)h(-1)) in the 16 588 patients randomized in the GUSTO V trial. Overall, the incidence of intracranial haemorrhage was similar in the two groups (0.6% vs 0.6%; OR 1.05, 95% CI 0.71, 1.56). The median (25th-75th) time from drug administration to intracranial haemorrhage was 5.5 (3.4-11) hours with combination therapy and 9.2 (5.9-22) hours with reteplase (P=0.048). Among the multivariable predictors of intracranial haemorrhage, only age showed a significant interaction with treatment effect (age per treatment interaction chi-square 4.60, P=0.032) with a lower risk of combination therapy for younger patients and a higher risk for the elderly. CONCLUSIONS: Although no additional risk of intracranial haemorrhage has been observed with combination therapy in the whole population, a significant age pertreatment interaction exists, with a lower risk with combination therapy in younger patients, and a higher risk in the elderly. PMID- 14563340 TI - Predictors of major bleeding in acute coronary syndromes: the Global Registry of Acute Coronary Events (GRACE). AB - AIMS: There have been no large observational studies attempting to identify predictors of major bleeding in patients with acute coronary syndromes (ACS), particularly from a multinational perspective. The objective of our study was thus to develop a prediction rule for the identification of patients with ACS at higher risk of major bleeding. METHODS AND RESULTS: Data from 24045 patients from the Global Registry of Acute Coronary Events (GRACE) were analysed. Factors associated with major bleeding were identified using logistic regression analysis. Predictive models were developed for the overall patient population and for subgroups of patients with ST-segment elevation myocardial infarction (STEMI), non-ST-segment elevation myocardial infarction (NSTEMI) and unstable angina. The overall incidence of major bleeding was 3.9% (4.8% in patients with STEMI, 4.7% in patients with NSTEMI and 2.3% in patients with unstable angina). Advanced age, female sex, history of bleeding, and renal insufficiency were independently associated with a higher risk of bleeding (P<0.01). The association remained after adjustment for hospital therapies and performance of invasive procedures. After adjustment for a variety of potential confounders, major bleeding was significantly associated with an increased risk of hospital death (adjusted odds ratio 1.64, 95% confidence interval 1.18, 2.28). CONCLUSIONS: In routine clinical practice, major bleeding is a relatively frequent non-cardiac complication of contemporary therapy for ACS and it is associated with a poor hospital prognosis. Simple baseline demographic and clinical characteristics identify patients at increased risk of major bleeding. PMID- 14563341 TI - Prognostic value and the changes of plasma levels of secretory type II phospholipase A2 in patients with coronary artery disease undergoing percutaneous coronary intervention. AB - AIM: To evaluate the serial changes of plasma secretory type II phospholipase A(2)(sPLA(2)), C-reactive protein (CRP) and cardiac injury markers in coronary artery disease (CAD) patients undergoing percutaneous coronary intervention (PCI) and their prognostic impacts. METHODS AND RESULTS: Plasma levels of sPLA(2), CRP, creatine kinase (CK), CK-MB and troponin-T were measured in 247 consecutive CAD patients receiving PCI procedure and 100 control subjects without CAD. In CAD group, serial blood samples were taken before coronary angiography, after coronary angiography, immediately after PCI, 24-h and 48-h after PCI. The sPLA(2)and CRP levels did not change after coronary angiography. The level of sPLA(2)significantly increased immediately after PCI. Creatine kinase and cardiac injury markers did not rise immediately after PCI, but elevated significantly at 24h after intervention. After a 2-year follow up, increased sPLA(2)(>450 ng/dl) after PCI, smoking and diabetes mellitus were the independent risk factors for subsequent coronary events (odds ratios 2.1, 2.3 and 3.1, respectively) in patients with CAD. CONCLUSION: The present study showed that PCI might cause immediate elevation of circulating levels of sPLA(2)following the mechanical disruption of coronary plaque, and the elevated level of sPLA(2)had significant prognostic impact. PMID- 14563342 TI - A polymorphism of the cholesteryl ester transfer protein gene predicts cardiovascular events in non-smokers in the West of Scotland Coronary Prevention Study. AB - AIM: The association of cholesteryl ester transfer protein (CETP) gene polymorphisms with risk of a cardiovascular event and whether any association was explained by an influence on high-density lipoprotein (HDL) levels or low-density lipoprotein (LDL) size was tested in the West of Scotland Coronary Prevention Study (WOSCOPS). Gene-smoking and gene-treatment interactions were investigated. METHODS AND RESULTS: Cases (n=498) and controls (n=1108) were typed for TaqIB, C( 631)A, C(-629)A, I405V and D442G CETP polymorphisms. Homozygotes for the TaqIB2 allele (B2B2) had a 30% reduced risk of a cardiovascular event (odds ratio [OR] 0.70, CI(95)0.51-0.96, P=0.03) compared to B1B1 homozygotes. Inclusion of HDL or LDL diameter in multivariate analysis only marginally attenuated the relationships. Non-smokers, but not smokers, showed a dose-dependent association of risk with TaqIB genotype. Treatment benefit was not significantly different in B1B1 (OR 0.71, pravastatin vs placebo), B1B2 (OR 0.68) and B2B2 (OR 0.61) individuals. The other CETP polymorphisms studied had no significant association with cardiovascular risk. Haplotype analysis did not add to the information given by the individual polymorphisms. CONCLUSION: The association between CETP TaqIB genotype and cardiovascular risk is primarily in non-smokers, is not fully explained by effects on HDL levels or LDL size, and the benefit of pravastatin treatment was not influenced by this polymorphism. PMID- 14563343 TI - Effect of pravastatin on LDL particle concentration as determined by NMR spectroscopy: a substudy of a randomized placebo controlled trial. AB - AIM: Recent data suggests that LDL particle concentration, as determined by Nuclear Magnetic Resonance (NMR) spectroscopy, may be associated with cardiovascular risk. We sought to determine the effect of randomization to pravastatin therapy on LDL particle concentration-NMR, among a primary prevention population. METHODS AND RESULTS: LDL particle concentration-NMR, LDL size-NMR, and standard chemical lipid parameters were measured at baseline and after 12 weeks among 500 individuals without overt coronary disease randomly allocated to pravastatin 40mg (n=256) or placebo (n=244). Randomization to pravastatin therapy caused a 19% reduction in median LDL particle concentration-NMR at 12 weeks, as compared to a 4.2% increase among those randomized to placebo (P<0.001 for pravastatin group compared to placebo). Pravastatin therapy caused a median 24.9% reduction in LDL cholesterol measured chemically compared to a 0.9% increase in the placebo group (P<0.001). Pravastatin therapy did not cause a significant change in median LDL size-NMR (0.5% increase in pravastatin group vs 0.0% in placebo group; P=0.25). The change in LDL particle concentration with pravastatin correlated inversely with baseline LDL size (r=-0.24; P<0.001) such that the largest reduction in LDL particle concentration-NMR was among those with the smallest LDL size-NMR at baseline (median% change =21.4% for tertile 1 of LDL size, 19.9% for tertile 2, and 16.5% for tertile 3; P=0.03). In contrast, pravastatin-induced changes in LDL cholesterol did not correlate with baseline LDL size-NMR (r=-0.05; P=0.47). CONCLUSION: Among individuals without overt hyperlipidemia or known coronary artery disease, randomized allocation to pravastatin (40mg) therapy for 12 weeks caused a reduction in LDL particle concentration-NMR, the magnitude of which was dependent on baseline LDL size-NMR. PMID- 14563344 TI - The 2373insG mutation in the MYBPC3 gene is a founder mutation, which accounts for nearly one-fourth of the HCM cases in the Netherlands. AB - AIMS: Hypertrophic cardiomyopathy (HCM) is caused by mutations in genes that encode sarcomeric proteins. In this study we investigated the involvement of the sarcomeric myosin binding protein C in the Dutch HCM population. METHODS AND RESULTS: We initially screened 22 Dutch index patients for mutations in the MYBPC3 gene, which revealed four different mutations in 14 patients. The 2373insG mutation was identified in 10 apparently unrelated patients. A subsequent screening for the 2373insG mutation in a group of another 237 unrelated HCM patients revealed 50 additional carriers of the same genetic defect. Genotyping with polymorphic repeat markers and intragenic SNPs of the 60 Dutch as well as two German and five North American 2373insG carriers indicated they all share the same haplotype. CONCLUSION: The 2373insG mutation accounts for almost one-fourth of all HCM cases in the Netherlands (60/259), which is predominantly present in the northwestern part of the country (22/66) and is a founder mutation probably originating from the Netherlands. PMID- 14563345 TI - Preconditioning and metabolic anti-ischaemic agents. AB - Preconditioning a powerful protective mechanism, is the response to transient ischemia and reperfusion. However, the best way to achieve total protection is to avoid ischemia altogether. Therefore prevention of ischemia and protection by preconditioning are differently mediated so that anti-ischemic agents may not precondition, whereas paradoxically pro-ischemic agents may precondition. Metabolically active agents such as glucose-insulin-potassium, trimetazidine and ranolazine that protect from ischemia, increase glucose metabolism relative to that of fatty acids. By promoting glycolysis they tend to close the ATP-dependent potassium channels that help to mediate preconditioning. By lessening the oxygen wasting effects of fatty acids, they are mitochondrial protective and oxygen sparing. These qualities should help in the therapy of myocardial ischemia and also heart failure. PMID- 14563346 TI - A critical amendment to the meta-analysis of clinical trials comparing thrombolysis with primary angioplasty. PMID- 14563349 TI - Chemokines and tuberculosis. AB - Mycobacterium tuberculosis is a respiratory pathogen responsible for tuberculosis. A primary pathologic feature of M. tuberculosis infection is the formation of a granuloma. Immune cells migrate to the lung and then through the lung to the site of infection to form a granuloma. This structure contains the infection, and is often maintained for a long period of time. The signals responsible for granuloma formation and maintenance are largely unknown. Since chemokines and chemokine receptors direct cells to specific sites within the tissues, it is plausible that these cells participate in granuloma formation. In this review, the current literature on chemokines and M. tuberculosis infection, as well as the specific role that tumor necrosis factor alpha (TNF-alpha) plays in granuloma formation and chemokine expression are discussed. PMID- 14563350 TI - The role of osteopontin in lung disease. AB - Osteopontin (Opn) is a multifunctional protein independently discovered by investigators from diverse scientific backgrounds and implicated in a broad array of pathological processes. Opn exists both intra- and extracellularly and in numerous pre- and post-translational isoforms. Structurally Opn resembles a matrix protein yet it has well-characterized cytokine like properties including the regulation of cellular migration and cell-mediated immunity. It has thus been classified as both a matricellular protein and a cytokine. Opn is among the most abundantly expressed proteins in a range of lung diseases and has been shown to regulate aspects of pulmonary granuloma formation, fibrosis, and malignancy. Future studies will explore the diagnostic and therapeutic potential of modulating the function of Opn in vivo. PMID- 14563351 TI - Immunomodulatory cytokines in asthmatic inflammation. AB - The development of asthmatic inflammation involves a complex array of cytokines that promote the recruitment and activation of a number of different immune cells. While factors involved in initiating and establishing inflammation are well characterized, the process by which this pro-inflammatory cascade is regulated is less well understood. The identification and characterization of immunomodulatory cytokines in asthma has been a difficult proposition. Many of the putative regulatory factors have pleiotropic bioactivities and have been characterized as pro-inflammatory in association with certain pathologic conditions. This chapter addresses the potential role of several endogenous factors which appear to attenuate asthmatic inflammation. Understanding the integration of these factors into the regulation of the inflammatory process will likely result in novel therapeutic approaches. PMID- 14563352 TI - Chemokines and cytokines: axis and allies in asthma and allergy. AB - Allergic asthma can be precipitated by many factors. For the atopic person, fungus, pollen, dust mites, cockroach antigens, and diesel exhaust are all agents that may trigger an allergic attack. Cytokines and chemokines are integral mediators of fungal asthma. From the earliest time points, they recruit and activate the cells required for the clearance of fungus as well as being critical factors involved in the immunopathology of this disease. In the final analysis, it is clear that these mediators can act to the benefit or the detriment of the host. PMID- 14563353 TI - Cytokine-directed therapies for the treatment of chronic airway diseases. AB - Multiple cytokines play a critical role in orchestrating and perpetuating inflammation in asthma and chronic obstructive pulmonary disease (COPD) and several specific cytokine and chemokine inhibitors now in development as future therapy for these diseases. Anti-IL-5 antibody markedly reduces peripheral blood and airway eosinophils, but does not appear to be effective in symptomatic asthma. Inhibition of IL-4 despite promising early results in asthma has been discontinued and blocking IL-13 might be more effective. Inhibitory cytokines, such as IL-10, interferons and IL-12 are less promising, as systemic delivery produces side effects. Inhibition of TNF-alpha may be useful in severe asthma and for treating severe COPD with systemic features. Many chemokines are involved in the inflammatory response of asthma and COPD and several small molecule inhibitors of chemokine receptors (CCR) are in development. CCR3 antagonists (which block eosinophil chemotaxis) and CXCR2 antagonists (which block neutrophil and monocyte chemotaxis) are in clinical development for asthma and COPD, respectively. Because so many cytokines are involved in asthma, drugs that inhibit the synthesis of multiple cytokines may prove to be more useful; several such classes of drug are now in clinical development and any risk of side effects with these non-specific inhibitors may be reduced by the inhaled route. PMID- 14563354 TI - Cytokine-mediated inflammation in acute lung injury. AB - Clinical acute lung injury (ALI) is a major cause of acute respiratory failure in critically ill patients. There is considerable experimental and clinical evidence that pro- and anti-inflammatory cytokines play a major role in the pathogenesis of inflammatory-induced lung injury from sepsis, pneumonia, aspiration, and shock. A recent multi-center clinical trial found that a lung-protective ventilatory strategy reduces mortality by 22% in patients with ALI. Interestingly, this protective ventilatory strategy was associated with a marked reduction in the number of neutrophils and the concentration of pro-inflammatory cytokines released into the airspaces of the injured lung. Further research is needed to establish the contribution of cytokines to both the pathogenesis and resolution of ALI. PMID- 14563356 TI - Neurotrophins and lung disease. AB - Neurotrophins are growth factors that exert multiple actions on neuronal and nonneuronal cells. Neurotrophin receptors are expressed on central and peripheral neurons, lymphocytes, monocytes, mast cells, and fibroblasts. In accordance with the distribution of their receptors, neurotrophins control the development and function of neurons and regulate inflammatory processes. Production of neurotrophins is altered in asthma, lung cancer, and pulmonary fibrosis. Evidence from animal models has implicated nerve growth factor (NGF) as a mediator of pulmonary inflammation, bronchoconstriction, and airway hyperreactivity, all of which are hallmarks of asthma. NGF regulates the growth of lung tumor cells and cultured lung fibroblasts. Thus neurotrophins, particularly NGF, are candidate molecules for regulating disease processes in asthma, lung cancer, and pulmonary fibrosis. PMID- 14563355 TI - Cytokine regulation of pulmonary fibrosis in scleroderma. AB - Pulmonary fibrosis occurs in up to 70% of scleroderma patients and progresses to cause severe restrictive lung disease in about 15% of patients. The mechanisms that cause pulmonary fibrosis in scleroderma remain incompletely understood. Increased amounts of mRNA or protein for multiple profibrotic cytokines and chemokines have been identified in lung tissue or broncholveolar lavage samples from scleroderma patients, when compared to healthy controls. These cytokines include transforming growth factor (TGF)-beta, connective tissue growth factor (CTGF), platelet-derived growth factor (PDGF), oncostatin M (OSM), monocyte chemotactic factor-1 and pulmonary and activation-regulated chemokine (PARC). Potential cellular sources of these profibrotic cytokines and chemokines in scleroderma lung disease include alternatively activated macrophages, activated CD8+ T cells, eosinophils, mast cells, epithelial cells and fibroblasts themselves. This review summarizes the literature on involvement of cytokines and chemokines in the development of pulmonary fibrosis in scleroderma. PMID- 14563357 TI - Neurotrophins in inflammatory lung diseases: modulators of cell differentiation and neuroimmune interactions. AB - Chronic inflammatory lung diseases represent a group of severe diseases with increasing prevalence as well as epidemiological importance. Inflammatory lung diseases could result from allergic or infectious genesis. There is growing evidence that the immune and nervous system are closely related not only in physiological but also in pathological reactions in the lung. Extensive communications between neurons and immune cells are responsible for the magnitude of airway inflammation and the development of airway hyperreactivity, a consequence of neuronal dysregulation. Neurotrophins are molecules regulating and controlling this crosstalk between the immune and peripheral nervous system (PNS) during inflammatory lung diseases. They are constitutively expressed by resident lung cells and produced in increasing quantities by immune cells invading the airways under inflammatory conditions. They act as activation, differentiation and survival factors for cells of both the immune and nervous system. This article will review the most recent data of neurotrophin signaling in the normal and inflamed lung and as yet unexplored, roles of neurotrophins in the complex communication within the neuroimmune network. PMID- 14563358 TI - Cloning and characterization of five cDNAs encoding peritrophin-A domains from the cat flea, Ctenocephalides felis. AB - Five cDNAs encoding peritrophin-A domains were identified as expressed sequence tags (ESTs) from flea hindgut and Malpighian tubule (HMT) cDNA libraries. The full-length cDNAs for each were subsequently isolated and sequenced. Three of the encoded proteins were similar to published peritrophin sequences, and thus were called "peritrophin-like", or PL1, PL2, and PL3. The other two sequences had similarity to both mucin and peritrophin proteins, and were called "mucin/peritrophin-like", or MPL1 and MPL2. The predicted protein sequences encoded by these cDNAs all contained a signal sequence and one or more peritrophin-A domains, which have been shown in other proteins to bind chitin. Aside from the peritrophin-A domains, the sequences shared little or no similarity to each other or to other proteins in the GenBank non-redundant database. The predicted protein sequences were variable in size, ranging in length from 81 to 453 amino acids. The two MPL proteins contained putative N linked and O-linked glycosylation sites, including a region of seven nearly perfect tandem repeats in the MPL1 protein sequence. Northern blot analysis of different flea lifestages and fed adult timepoints showed distinct mRNA expression patterns for each gene, although all five transcripts were primarily or exclusively detected in the HMT tissues in adults. The PL1 protein was detected by immuno-blot in soluble and insoluble protein extracts from unfed and fed adult fleas. The PL1 protein from the insoluble fractions appeared to be approximately 1 kDa larger than the PL1 protein from the soluble protein fractions. Immunohistochemistry performed on flea thin sections revealed that the PL1 protein was detected in the Malpighian tubules, hindgut, rectum, and trachea. Unpurified native PL1 protein from both soluble and insoluble protein fractions was tested for chitin-binding activity but did not bind to chitin under the conditions tested. These results show that the flea peritrophin-like proteins may have biological functions that are distinct from the peritrophic matrix and from the binding of chitin. PMID- 14563359 TI - Distinct LPS-induced signals regulate LPS uptake and morphological changes in medfly hemocytes. AB - Recently we demonstrated that lipopolysaccharide (LPS) promotes activation of the Ras/ERK cascade in medfly hemocytes and that phagocytosis of Escherichia coli by insect hemocytes is mediated by an integrin-dependent process via the activation of FAK/Src complex (J Biol Chem 273 (1998) 14813; FEBS Letters 496 (2001) 55). In the current study we wanted to further elucidate the effects of LPS on medfly hemocytes, in order to better understand the regulation of the evolutionary conserved signaling mechanisms between insects and mammals. We initially observed that different stimuli, including LPS, E. coli, RGD, fibronectin and heat shock activate hemocyte ERK. The response of hemocytes to these stimuli denoted that hemocyte ERK is evidently stimulated by at least an LPS receptor and via an integrin-mediated process. The medfly hemocytes respond to LPS by changing their morphology, inducing the activation of several signaling pathways, including Ras/MEK/ERK, PI-3K/ERK and Rho pathways and contributing to LPS uptake. Experiments based on inhibitors of specific signaling pathways, such as manumycin A, toxin A, U0126, PD98059 and wortmannin revealed that Ras, MEK and PI-3K are involved in the activation of ERK. Whether PI-3K is an intermediate of Ras/MEK/ERK pathway or activates ERK via other signaling pathway it remains to be elucidated. ERK is not activated via Rho pathway, denoting that Rho may not be an upstream effector molecule of ERK pathway. Regarding the role(s) that these kinases play in hemocytes, it can be suggested that PI-3K and Rho GTPases can modulate hemocyte shape changes, whereas ERK, Ras and MEK cannot. In addition, PI 3K as well as Ras and MEK through ERK activation participate in LPS endocytosis. Therefore, PI-3K shares a dual role; it is involved both in cell shape changes and in LPS endocytosis. Since ERK activation appears to be independent of the integrity of actin filaments, as cytochalasin D and latrunculin A did not block ERK activation, it can be concluded that LPS endocytosis is independent of actin cytoskeleton remodeling as is the case in mammalian systems. PMID- 14563360 TI - Action pattern, specificity, lytic activities, and physiological role of five digestive beta-glucanases isolated from Periplaneta americana. AB - Three laminarinases (LAM, LIC 1, and LIC 2) and two cellulases (CEL 1 and CEL 2) were purified to homogeneity from Periplaneta americana midguts. These beta glucanases are secreted by salivary glands, stabilized by calcium ions, and have pH optima around 6. LAM (46 kDa) is active only on laminarin, native or with oxidized ends, and so it is an endo-beta-1,3-glucanase (EC 3.2.1.39). It processively releases mainly glucose from laminarin and shows lytic activity on fungal cells. LIC 1 (25 kDa) is an endo-beta-1,3(4)-glucanase (EC 3.2.1.6.), because it cleaves internal bonds on both laminarin and lichenin. LIC 1 lyses fungal cells and apparently have high affinity for sequences of cellotetraoses linked by beta-1,3 links, releasing cellotetraose from lichenin. The reaction catalyzed by LIC 1 is not in rapid equilibrium, as suggested by activity-pH data. These data also showed that a group in LIC 1 with pK=4.9 is necessary for substrate binding. LIC 2 (23 kDa) seems to be similar to LIC 1. The laminarinases are inactivated by carbodiimide, suggesting the presence of a carboxyl group involved in catalysis. LAM and LIC 2 are inhibited by excess laminarin as substrate. CEL 1 (72 kDa) and CEL 2 (73 kDa) quickly decrease the molecular weight of lichenin used as substrate. Therefore, they are endo-beta-1,4 glucanases (EC 3.2.1.4). Both CEL 1 and CEL 2 are also active on crystalline cellulose. The specificities of P. americana beta-glucanases agree with the omnivorous detritus-feeding habit of this insect, as they are able to attack plant (CEL 1, CEL 2, LIC 1 and LIC 2) and fungal (LIC 1 and LAM) cell walls. PMID- 14563361 TI - An arthropod defensin expressed by the hemocytes of the American dog tick, Dermacentor variabilis (Acari: Ixodidae). AB - Both soluble and cell-mediated components are involved in the innate immune response of arthropods. Injection of Borrelia burgdorferi, the Lyme disease agent, results in the secretion of defensin into the hemolymph of the ixodid tick, Dermacentor variabilis. The presence of the peptide is observed as early as 15 min post-challenge and remains present through 18 h post-challenge. As observed in insects and soft ticks, the transcript for defensin is detected as early as 1 h post-challenge in D. variabilis. RT-PCR resulted in an amplicon of 624 bp with a 225 bp region that translates to a 74 amino acid preprodefensin. The defensin encoding region was amplified, cloned and sequenced from the hemocytes. It appears as though defensin is stored in the granulocytes of the hemolymph and secreted into the hemolymph upon bacterial insult. The role of defensin as a contributing factor in determining vector competency is discussed. PMID- 14563362 TI - Blood meal induces global changes in midgut gene expression in the disease vector, Aedes aegypti. AB - Blood feeding is an essential developmental process for many arthropods and plays a significant role in disease transmission. Understanding physiological responses in the midgut is important because it is the primary site of blood meal digestion and pathogenic infection. Processes that occur in the midgut in response to a blood meal have been studied but are poorly understood. Here, we use cDNA microarrays to examine midgut gene expression on a global level in response to blood feeding to assist in unraveling these processes. We have developed Aedes aegypti microarrays consisting of clones obtained from an expressed sequence tag project. Individual clones were amplified by polymerase chain reaction and printed onto glass slides. These microarrays were used to study the effects of a blood meal on midgut gene expression over a 72-h time course. As a result, a number of genes involved in processes such as nutrient uptake and metabolism, cellular stress responses, ion balance, and PM formation, as well as a number of unknown genes were induced or repressed in response to a blood meal based on this microarray data. PMID- 14563363 TI - Characterization of recombinant and native Ih-channels from Apis mellifera. AB - Recently, a novel class of genes coding for Ih-channels has been identified in several vertebrates and invertebrates. We isolated a cDNA (AMIH) encoding a putative member of these ion channels from Apis mellifera heads by means of polymerase chain reaction and homology screening. High similarity (88% identical amino acids) to the putative Drosophila melanogaster Ih-channel suggests that the Apis cDNA codes for a hyperpolarization-activated and cyclic nucleotide-gated channel. Functional expression of recombinant AMIH in HEK293 cells gave unitary currents that were preferentially selective for potassium over sodium ions and were activated by hyperpolarizing voltage steps. Cyclic nucleotides shifted the voltage activation curve to more positive membrane potentials. The current kinetics, activation by hyperpolarizing voltage steps and modulatory influence of cyclic nucleotides properties closely resemble those of mammalian Ih-channels. RT PCR analysis showed pronounced mRNA expression in the antennae, head and body of Apis mellifera. Investigation of hyperpolarization-activated currents in olfactory receptor neurons (ORNs) in a primary cell culture of Apis mellifera antennal cells revealed activation properties similar to the heterologously expressed Ih-channel. By in-situ hybridization and immunohistochemistry, expression of AMIH was seen in olfactory receptor neurons of the bee antennae. We conclude that AMIH is the ion channel responsible for the hyperpolarization activated currents in olfactory receptor neurons of bee. PMID- 14563364 TI - Transcriptome identification of putative genes involved in protein catabolism and innate immune response in human body louse (Pediculicidae: Pediculus humanus). AB - Genomics information relating to human body lice is surprisingly scarce, and this has constrained studies of their physiology, immunology and vector biology. To identify novel body louse genes, we used engorged adult lice to generate a cDNA library. Initially, 1152 clones were screened for inserts, edited for removal of vector sequences and base pairs of poor quality, and viewed for splicing variations, gene families and polymorphism. Computational methods identified 506 inferred open reading frames including the first predicted louse defensin. The inferred defensin aligns well with other insect defensins and has highly conserved cysteine residues, as are known for other defensin sequences. Two cysteine and five serine proteinases were categorized according to their inferred catalytic sites. We also discovered seven putative ubiquitin-pathway genes and four iron metabolizing deduced enzymes. Finally, glutathione-S-transferases and cytochrome P450 genes were among the detoxification enzymes found. Results from this first systematic effort to discover human body louse genes should promote further studies in Phthiraptera and lice. PMID- 14563365 TI - The DNA binding of insect Fork head factors is strongly influenced by the negative cooperation of neighbouring bases. AB - The Drosophila melanogaster Fork head and Bombyx mori SGF1/Fork head proteins are key regulators of tissue specific gene expression in the modified larval labial glands. Here we use the competitive electrophoretic mobility shift assay to create a detailed Fork head binding matrix and we investigate some unusual features of the Fork head interaction with DNA. We found that the Fork head-DNA interaction is context dependent--the binding specificity of the protein is partly determined by specific combinations of neighbouring bases. Although the total number of the sub-optimal dinucleotide steps is not high, the negative cooperation of neighbouring bases significantly contributes to the overall binding site specificity. Our results allow efficient recognition of insect Fork head binding sites and we show that the putative Fork head cognate elements preferentially accumulate in the near upstream region of genes abundantly expressed in the labial gland. PMID- 14563366 TI - Proventriculus (cardia) plays a crucial role in immunity in tsetse fly (Diptera: Glossinidiae). AB - Fat body and hemocytes play a central role in cellular and humoral responses for systemic infections in invertebrates, similar to the mammalian liver and blood cells. Epithelial surfaces, in particular the midgut, participate in the initial local immune responses in order to aid in the generation of the terminal cytotoxic molecules that mediate non-self recognition. Here, we describe for the first time the immune responses of a cluster of cells at the foregut/midgut junction--known as proventriculus (cardia) in the medically and agriculturally important insect, tsetse fly (Diptera: Glossinidae). We provide evidence for the transcriptional induction of the antimicrobial peptides attacin and defensin as well as for the reactive nitrogen intermediate (RNI) nitric oxide synthase (NOS) upon microbial challenge by either microinjection or feeding. Proventriculus from immune challenged flies also has higher NOS and nitric oxide (NO) activities as well as increased levels of the reactive oxygen intermediate (ROI), hydrogen peroxide (H2O2). In several vector pathogen systems, including tsetse flies and African trypanosomes, stimulation of systemic responses prior to pathogen acquisition has been shown to reduce disease transmission. Furthermore, the induction of systemic immune responses has been documented while pathogens are still differentiating within the midgut environment. While evidence for a close molecular communication between the local and systemic responses is accumulating, the molecular signals that mediate these interactions are at present unknown. Reactive intermediates such as NO or H2O2 may function as immunological signals for mediating the molecular communication between the different insect compartments. We discuss the putative role of the proventriculus in invertebrate immunity and specifically speculate on its significance for trypanosome transmission in tsetse. PMID- 14563367 TI - A cationic peptide consists of ornithine and histidine repeats augments gene transfer in dendritic cells. AB - Condensing the plasmid with high molecular weight cationic polymers such as poly L-lysine (PLL) and poly-L-ornithine (PLO) can enhance antigen-specific immunity generated from genetic vaccination with naked DNA encoding antigens. While these high molecular weight polymers are clearly effective in transfection experiments, clinical applications are limited by their physical heterogeneity and toxicity. Three chemically defined low molecular weight cationic peptides, K(16), K(10)H(6), and O(10)H(6), were examined in the context of DNA binding, toxicity, and efficiency of gene transfer in dendritic cells (DC). The results showed that while all three peptides can bind to a plasmid encoding a reporter gene with similar efficiency, in vitro transfection with DNA complexed with O(10)H(6) complexed resulted in the highest level of gene expression. Moreover, free O(10)H(6) was not toxic to DC, while the lysine-based peptides caused significant cell death in DC cultures. We also showed that DC transfected ex vivo with DNA complexed with O(10)H(6) was capable of eliciting antigen-specific INFgamma production in vivo. Taken together, these results indicate ornithine and histidine repeats are suitable building blocks of non-viral gene transfer vector for DC. PMID- 14563368 TI - Molecular signatures of anti-nuclear antibodies--contribution of heavy chain framework residues. AB - It is clear that besides the CDR residues, framework residues (particularly those on FR1 and FR3) can contribute towards antigen reactivity. This study was designed to compare the immunoglobulin heavy chain FR regions of anti-nuclear antibodies (comprised of 142 anti-ssDNA, 103 anti-dsDNA and 23 anti-nucleosome Abs) with those of non-nuclear antibodies (N=165), all drawn from the GenBank. The anti-nuclear antibodies depicted residue-usage differences that reached statistical significance in their FR1 (at H1 and H29), FR2 (at H40), and FR3 (at H69, H73, H76, H80 and H87) regions. Interestingly, these residue-usage differences were intimately linked to differences in the usage frequencies of different V(H) germline genes between the different groups of antibodies; thus, whereas J558.4, J558.47, J558.m, and 7183.9 germline genes were over-utilized among the ANAs, J558.17, V130, V(H) 36-60 and VGk1a were over-represented among the non-ANAs. In conclusion, although the framework regions of ANAs exhibited distinctly different residue-usage patterns, they may simply be markers of associated germline-encoded CDR differences that appear to have been co-selected. Further studies are warranted to ascertain if any of the observed framework residue differences do actually contribute to nuclear antigen reactivity. PMID- 14563369 TI - The polymorphisms of intron1 sequences of HLA-A and -B. AB - Human leucocyte antigen (HLA) class I antigens are highly polymorphic membrane glycoproteins present on most nucleated cells. The polymorphism region is mainly located in exons 2 and 3 of HLA class I genes, which is flanked by introns 1 and 3. Thus, the sequence information of introns 1 and 3 is important for the genotyping of HLA. However, the information about them has not been extensively reported by now. In this work, the intron 1 sequences of HLA-A and -B of 51 standard genomic DNAs provided by the 13th International Histocompatibility Work Group (IHWG) were cloned and sequenced. The polymorphism of the intron sequences were also analyzed by the software Clustal W 1.82. Some sequences were chosen to compare with the standard sequences published in National Center for Biotechnology Information (NCBI). The comparison showed that all the sequences matched exactly with the standard sequences. The knowledge of intron 1 sequences could be very important not only for developing DNA-based typing strategies for the HLA-A and -B alleles but also for establishing an understanding of the evolutionary mechanisms involved in the polymorphism generation of HLA class I alleles. PMID- 14563370 TI - Germline transcripts of immunoglobulin light chain variable regions are structurally diverse and differentially expressed. AB - The murine pre-B cell line R2-bfl, which can be induced to differentiate in vitro, was used to study germline transcription of variable regions of the light chain loci. RNA from these cells was subjected to a 3'-RACE and germline transcripts from 17 individual Vkappa gene segments belonging to 12 Vkappa families were characterized. Germline transcripts of all three Vlambda regions were similarly analyzed. The synchronous differentiation of R2-bfl cells was then used to investigate the order of appearance of germline transcripts of the V and JC clusters of both light chain loci. This was taken as indicator for accessibility of a particular locus to rearrangement. Germline transcripts of the JCkappa cluster and the Vkappa family most proximal to JCkappa was detectable already at day 0, while transcripts of the most distal Vkappa family became apparent after initiation of differentiation at day 1. Transcripts of the JClambda cluster could be found at day 2, whereas transcripts of the Vlambda region were already present at day 1. Thus, the lambda locus becomes accessible to rearrangement later during development than kappa, confirming and extending our previous findings. The V and JC clusters open at the same stage of development although slight asynchronicities were found for the Vlambda and the distal Vkappa gene segments. PMID- 14563371 TI - Anti-cardiolipin/beta-2 glycoprotein activities co-exist on human anti-DNA antibody light chains. AB - We have recently shown that the human anti-DNA antibodies B3 and 33H11 also bind cardiolipin and that the anti-autoantigen activity resides predominantly on their lambda light chains. We now show that the two auto-antibodies possess strong reactivity to the plasma-protein 2-Glycoprotein I (beta2-GPI) also. Utilizing chain shuffling experiments involving an unrelated anti-p185 antibody 4D5 with insignificant reactivity to cardiolipin or to beta2-GPI, we now demonstrate that hybrid Fabs with constituent light chain, but not the heavy chain, of B3 or 33H11, exhibit anti-cardiolipin activity. Furthermore, the constructs possessing the auto-antibody-derived light chain also exhibited significant reactivity to beta2-GPI. The results suggest that anti-DNA, anti-cardiolipin and anti-beta2-GPI activities co-exist on the light chains of the antibodies studied and, importantly, these activities could be transferred to antibody constructs by their light chains alone. Computer-generated models of the three-dimensional structures of the auto-antibodies and their hybrids, suggest predominant interaction of their light chains with domain IV of beta2-GPI. PMID- 14563372 TI - Phosphatases in concert with kinases set the gain for signal transduction through the T cell receptor. AB - The 'tunable activation thresholds' model for signal transduction through the T cell receptor (TCR)/CD3 signaling complex proposes that rapid cycles of phosphorylation and dephosphorylation are integral to regulating the frequency of protein-protein interaction, thus having considerable influence over the activation of downstream signaling pathways. Co-temporal activation of kinases and phosphatases could serve to modulate the ongoing signaling response, depending on the relative balance of their opposing enzymatic activities. Although recent reports have addressed the mechanisms by which specific kinase/phosphatase pairs contribute to the initiation and termination of signaling, we sought a more global understanding of the ability of the kinase/phosphatase balance to regulate, or "tune", the very proximal steps of TCR signaling in primary human T cells. Herein, we provide biochemical evidence that phosphotyrosine induction via the TCR is subject to fine-tuning based on the overall activity of kinases and phosphatases relative to one another, leading to cycles of phosphorylation and dephosphorylation, with implications for developing the next generation of immunotherapeutic agents. PMID- 14563373 TI - Regulation of CXCR4 expression in human T lymphocytes by calcium and calcineurin. AB - Principally expressed on the surface of T lymphocytes, the chemokine and HIV receptor CXCR4 has been shown to serve key roles in both chemotaxis and HIV-1 entry into T cells. Understanding the regulation of CXCR4 expression is therefore of paramount importance to further elucidating its endogenous role and contributions to HIV-1 pathogenesis. Using an RNase protection assay (RPA), we have demonstrated that mitogenic stimulation of purified human peripheral blood T lymphocytes (PBL) decreased CXCR4 mRNA relative to unstimulated controls in a calcineurin-dependent manner; an expression pattern mimicked by the chemokine receptor CCR7. A change in transcriptional activity, not in mRNA stability, was required for control of CXCR4 and CCR7 expression. Changes in CXCR4 mRNA expression translated into a stimulation- and calcineurin-dependent decrease in cell surface CXCR4 expression. We have previously demonstrated that CXCR4 mRNA and protein is regulated by cAMP; here we show that calcium and calcineurin signaling pathways modify cAMP-driven changes. Moreover, we provide data supporting a role for the transcription factor YY1 in calcineurin-dependent regulation of CXCR4 expression. PMID- 14563375 TI - A review of functional neuroimaging studies of vagus nerve stimulation (VNS). AB - Vagus nerve stimulation (VNS) is a new method for preventing and treating seizures, and shows promise as a potential new antidepressant. The mechanisms of action of VNS are still unknown, although the afferent direct and secondary connections of the vagus nerve are well established and are the most likely route of VNS brain effects. Over the past several years, many groups have used functional brain imaging to better understand VNS effects on the brain. Since these studies differ somewhat in their methodologies, findings and conclusions, at first glance, this literature may appear inconsistent. Although disagreement exists regarding the specific locations and the direction of brain activation, the differences across studies are largely due to different methods, and the results are not entirely inconsistent. We provide an overview of these functional imaging studies of VNS. PET (positron emission tomography) and SPECT (single photon emission computed tomography) studies have implicated several brain areas affected by VNS, without being able to define the key structures consistently and immediately activated by VNS. BOLD (blood oxygen level dependent) fMRI (functional magnetic resonance imaging), with its relatively high spatio-temporal resolution, performed during VNS, can reveal the location and level of the brain's immediate response to VNS. As a whole, these studies demonstrate that VNS causes immediate and longer-term changes in brain regions with vagus innervations and which have been implicated in neuropsychiatric disorders. These include the thalamus, cerebellum, orbitofrontal cortex, limbic system, hypothalamus, and medulla. Functional neuroimaging studies have the potential to provide greater insight into the brain circuitry behind the activity of VNS. PMID- 14563374 TI - Major linear IgE epitopes of mountain cedar pollen allergen Jun a 1 map to the pectate lyase catalytic site. AB - Resolution of the 3D structures and IgE epitopes of allergens may identify common or conserved features of allergens. Jun a 1, the predominant allergen in mountain cedar pollen, was chosen as a model for identifying common structural and functional features among a group of plant allergens. In this study, synthetic, overlapping peptides of Jun a 1 and sera from patients allergic to mountain cedar pollen were used to identify linear epitopes. A 3D model of Jun a 1 was produced using the Bacillus subtiles pectate lyase (PL) as a template and validated with biophysical measurements. This allowed mappings of four IgE binding sites on Jun a 1. Two of the epitopes mapped to turns or loops on the surface of the model structure. The other two epitopes mapped to the beta-sheet region, homologous to the catalytic site of PL. This region of Jun a 1 is highly conserved in the group 1 allergens from other cedar trees as well as microbial PLs. The finding that two out of three major IgE epitopes map to highly conserved catalytic regions of group 1 cedar allergens may help to explain the high degree of cross-reactivity between cedar pollen allergens and might represent a pattern of reactivity common to other allergens with catalytic activity. PMID- 14563377 TI - Hypothalamo-pituitary-adrenal function in patients with depressive disorders is correlated with baseline cytokine levels, but not with cytokine responses to hydrocortisone. AB - Dysfunction of the hyopthalamo-pituitary adrenal (HPA) system is frequently found in major depression. In addition, signs of non-specific inflammatory system activation have been reported. However, very little is known about interactions between the HPA and immune systems in depressive patients. To assess HPA system function, we performed a combined dexamethasone suppression and corticotropin releasing hormone stimulation (DEX/CRH) test in 14 depressive patients. Moreover, baseline nocturnal plasma levels of the inflammatory cytokines interleukin-6 (IL 6) and tumor necrosis factor (TNF)-alpha were measured. In addition, the system was challenged with an intraveneous pulsatile injection of hydrocortisone (1 mg/kg body weight in total) and again cytokine levels were measured across one night. Baseline TNF-alpha levels were negatively correlated with the amount of ACTH released upon CRH stimulation during the DEX/CRH test. Acute hydrocortisone administration suppressed TNF-alpha and IL-6 levels independently of baseline HPA system activity. We conclude that chronic HPA system overactivity in depressed patients might compromise the production of inflammatory cytokines under baseline conditions. However, the responsivity of the cytokine production to acutely administered glucocorticoids does not seem to correlate with the state of the HPA system. PMID- 14563376 TI - Association between IL-1beta -511C/T and IL-1RA (86bp)n repeats polymorphisms and schizophrenia. AB - It has been established that cytokines play a critical role in the regulation of the CNS and recent studies have suggested that dysfunctions of both pro inflammatory (IL-1beta, IL-6, and TNF-alpha) and anti-inflammatory (IL-1RA and IL 10) cytokines could be involved in the pathophysiology of schizophrenia. Previous studies have reported that functional polymorphisms in some cytokines genes may have important regulatory effects on such system. Therefore, the aim of the present study was to explore the possible role of the IL-1beta -511C/T and IL-1RA (86bp)(n) repeats polymorphisms in schizophrenia. A case control association study comparing genotype and allele frequencies in 346 northen Italian subjects (169 schizophrenic patients and 177 unrelated healthy volunteers) was performed. The frequencies of IL-1beta -511C and IL-1RA allele 1 (86bp)(4) are significantly higher in schizophrenic patients compared to controls (IL-1beta -511 P=0.047; IL 1RA (86bp)(n) P=0.002). Moreover our data show a protective effect of the IL-1RA allele 2 (86bp)(2) against schizophrenia (OR=0.59 95%CI:0.388-0.910; P=0.016) and this effect is enhanced by the concomitant presence of IL-1beta -511T (OR=0.48 95%CI:0.30-0.76; P=0.002). Our findings support the hypothesis that genetically determined changes in IL-1 metabolism regulation may contribute to the pathogenesis of schizophrenia confirming a role of IL-1 gene cluster in disease susceptibility. PMID- 14563378 TI - Relationship between dexamethasone-inhibited lysozyme activity in peripheral mononuclear leukocytes and the cortisol and glucocorticoid receptor response to dexamethasone. AB - The assessment of lysozyme activity in mononuclear leukocytes (MNLs) following the in vitro administration of dexamethasone (DEX) provides a measure of peripheral glucocorticoid sensitivity. The goal of the present study was to determine the relationship between the IC(50) of lysozyme activity following such challenge, and the cortisol response to oral administration of 0.50 mg DEX in 18 healthy subjects. The results demonstrated a robust association between the IC(50) and both cortisol decline and percent suppression of cortisol in response to low-dose DEX. However, this measure was uncorrelated with pre or post DEX cortisol levels or GR number. The high correlation between the inhibitory effect of DEX on lysozyme synthesis and two measures reflecting cortisol suppression in response to oral DEX reflects the similarities of GC responsiveness in both in vivo and in vitro models, and suggests that the in vitro assessment of lysozyme activity in MNLs may be useful in the study of neuropsychiatric or other clinical disorder. PMID- 14563379 TI - Specific executive/attentional deficits in patients with schizophrenia or bipolar disorder who have a positive family history of psychosis. AB - Neurocognitive impairments are well documented in patients with schizophrenia and their healthy first-degree biological relatives. Less is known about neuropsychological performance in bipolar disorders, but some studies indicate that, compared to schizophrenia, bipolar disorder displays a similar profile pattern with less severe deficits. The genetic and environmental contributions to the development of neurocognitive deficits are also unclear. This study explored the effect of a family history (FH) of psychotic disorders in first-degree relatives on a variety of cognitive domains (abstraction and flexibility, verbal fluency, verbal memory, motor activity and visual-motor processing/attention) in 30 patients with schizophrenia, and 24 type I bipolar patients. After adjusting the results for age, gender, education level and pre-morbid intelligence, patients with schizophrenia or bipolar disorder with positive FH (n=18) performed significantly worse than patients with negative FH (n=36) on the visual-motor processing/attention domain. These findings were independent of the specific diagnosis. Moreover, when logistic regression analysis was performed, poor Digit Symbol performance was the only predictor of belonging to the positive FH group. Our results are compatible with the existence of some common genetic factors between the illnesses, as well as the involvement of identical, or at least similar, disordered brain systems in both disorders. These findings are discussed within the context of the continuum model of psychosis. PMID- 14563381 TI - Fast psychomotor functioning in underweight anorexia nervosa patients. AB - To explore whether underweight anorexia nervosa patients show psychomotor differences relative to normal controls, 32 female hospitalised patients, aged between 14 and 25 years, were compared with 32 healthy, normal weight controls, matched for sex, age and educational level. Using computerised analysis of writing and drawing behaviour, reaction times and movement times and their different components were analysed, while cognitive and motor demands were manipulated in five drawing and copying tasks. Anorexia nervosa patients were, compared to normal weight controls, significantly faster in a drawing task and showed shorter reaction times in copying tasks. Movement times did not differ significantly between the two groups. In the most complex copying task, a significant group x complexity interaction for reaction time (patients shorter) and reinspection time (patients longer) was found. Patients also made more errors than controls. The finding of a consistent pattern of shorter reaction times in underweight anorexia nervosa patients seems to run contrary to previous findings of disturbed cognitive functioning (i.e. impaired attention) in these patients. The differences seem to be related to cognitive factors more than to motor (executive) components. PMID- 14563380 TI - The distribution and morphology of prefrontal cortex pyramidal neurons identified using anti-neurofilament antibodies SMI32, N200 and FNP7. Normative data and a comparison in subjects with schizophrenia, bipolar disorder or major depression. AB - Alterations in the density and size of pyramidal neurons in the prefrontal cortex have been described in schizophrenia and mood disorder. However, the changes are generally modest and have not always been replicated. We investigated the possibility that specific pyramidal neuron sub-populations, defined by their immunoreactivity with the anti-neurofilament antibodies SMI32, N200, and FNP7, are differentially affected in these disorders. First, we assessed the distribution and characteristics of pyramidal neurons labelled by the antibodies in the human dorsolateral prefrontal cortex (Brodmann areas 9, 32, 46), using single and double label immunocytochemistry and immunofluorescence. Three largely separate sub-populations of pyramidal neurons were identified, although with more substantial overlap between SMI32- and FNP7-positive neurons in lamina V. We then determined the density, size and shape of the three pyramidal neuron sub populations in area 9 in patients with schizophrenia, bipolar disorder, or major depressive disorder, compared to controls (n=15 in each group). We found a lower density of lamina III N200-positive neurons in major depressive disorder than in schizophrenia or bipolar disorder. There were no other overall differences in neuronal density, or in neuronal size or shape, although a planned secondary analysis supported the previously reported decrease of neuronal size in lamina V in bipolar disorder. In summary, our study illustrates a conceptual and methodological approach which may be of value for investigating the differential neuropathological involvement of pyramidal neuron sub-populations. However, we found no clear evidence that the prefrontal neuropathology of schizophrenia or mood disorders preferentially affects SMI32-, N200- or FNP7-immunoreactive pyramidal neurons. PMID- 14563382 TI - Development of a standardized training program for the Hamilton Depression Scale using internet-based technologies: results from a pilot study. AB - Poor inter-rater reliability is a major concern, contributing to error variance, which decreases power and increases the risk for failed trials. This is particularly problematic with the Hamilton Depression Scale (HAMD), due to lack of standardized questions or explicit scoring procedures. Establishing standardized procedures for administering and scoring the HAMD is typically done at study initiation meetings. However, the format and time allotted is usually insufficient, and evaluation of the trainee's ability to actually conduct a clinical interview is limited. To address this problem, we developed a web-based, interactive rater education program for standardized training to diverse sites in multi-center trials. The program includes both didactic training on scoring conventions and live, remote observation of trainees applied skills. The program was pilot tested with nine raters from a single site. Results found a significant increase in didactic knowledge pre-to-post testing, with the mean number of incorrect answers decreasing from 6.5 (S.D.=1.64) to 1.3 (S.D.=1.03), t(5)=7.35, P=0.001 (20 item exam). Seventy-five percent of the trainees' interviews were within two points of the trainer's score. Inter-rater reliability (intraclass correlation) (based on trainees actual interviews) was 0.97, P<0.0001. Results support the feasibility of this methodology for improving rater training. An NIMH funded study is currently underway examining this methodology in a multi-site trial. PMID- 14563383 TI - 2-OH-estradiol, an endogenous hormone with neuroprotective functions. AB - We compared the neuroprotective effects of the catecholestrogen 2-hydroxy estradiol (2-OH-E(2)) to the actions of 17-beta-estradiol (E(2)), since catecholestrogens have been clinically implicated in the pathophysiology of major depression and other psychiatric diseases. Using the hippocampal HT22 cell line as a well-established in vitro model system, we here show that the extent of the neuroprotective effects of 2-OH-E(2) was significantly increased compared to the physiological estrogen E(2) at equimolar concentrations after a toxic challenge with hydrogen peroxide. Statistically significant effects of neuroprotection as measured by survival of HT22 cells were detectable at concentrations of 1 and 10 microM of 2-OH-E(2) or E(2). Studies on the time-dependence of the evoked reactions showed that a pre-incubation and a post-incubation up to 30 min with a dose of 10 microM of 2-OH-E(2) resulted in a significant decrease in cell death after incubation with hydrogen peroxide if compared to E(2). Further characterization of the effects in rat brain homogenates with an assay for the induction of cellular lipid peroxidation (LPO) revealed, that 2-OH-E(2) was more effective in the reduction of LPO than E(2) in equimolar concentrations. This indicates a pharmacologically relevant effect of this hormone metabolite and a mechanism of action, which does not involve the classical estrogen receptor. In conclusion, the catecholestrogen 2-OH-E(2) induces increased neuroprotective actions in comparison to the major physiological estrogen E(2), suggesting a clinically relevant physiological function of catecholestrogens during health and disease. PMID- 14563384 TI - Treatment of depression with the CRH-1-receptor antagonist R121919: endocrine changes and side effects. AB - A dysregulation of the hypothalamus-pituitary-adrenocortical (HPA) system has been hypothesized to account for a myriad of cardinal symptoms of affective disorders. Specifically, increased CRH signalling via CRH type 1 receptors is thought to be an important factor in the pathogenesis of major depression and anxiety disorders. Consequently, a number of drugs have been developed in order to target the postulated increase in CRH/CRH 1 receptor signalling. One of these compounds, R121919, binds with high affinity to CRH1 receptors antagonising the action of CRH. R121919 was recently tested in an open-label study conceptualized as a safety and tolerability study. As part of this study, a thorough endocrine evaluation and detailed clinical laboratory analysis were assessed several times during 30 days of treatment with two different dose regimens of R121919 (5-40 mg vs. 40-80 mg) in 24 patients with a major depressive episode. During treatment with the experimental drug no serious side effects were noted. In particular, there were no adverse effects or impairment of the hypothalamic-pituitary-gonadal system, the hypothalamic-pituitary-thyroid axis, the renin-angiotensin system, prolactin or vasopressin secretion. Furthermore, no changes in the serum corticotropin and cortisol concentrations and in the responsivity of corticotropin and cortisol following a CRH stimulation test were noted. No effects of R121919 on clinical laboratory parameters including liver enzymes, EEG and ECG were observed. These results encourage the development of other CRH-1-R antagonists as a novel class of antidepressive drugs. PMID- 14563385 TI - Age and disease severity predict choice of atypical neuroleptic: a signal detection approach to physicians' prescribing decisions. AB - OBJECTIVE: We used a novel application of a signal detection technique, receiver operator characteristics (ROC), to describe factors entering a physician's decision to switch a patient from a typical high potency neuroleptic to a particular atypical, olanzapine (OLA) or risperidone (RIS). METHODS: ROC analyses were performed on pharmacy records of 476 VA patients who had been treated on a high potency neuroleptic then changed to either OLA or RIS. RESULTS: Overall 68% patients switched to OLA and 32% to RIS. The best predictor of neuroleptic choice was age at switch, with 78% of patients aged less than 55 years receiving OLA and 51% of those aged greater than or equal to 55 years receiving OLA (chi(2)=38.2, P<0.001). Further analysis of the former group indicated that adding the predictor of one or more inpatient days to age increased the likelihood of an OLA switch from 78% to 85% (chi(2)=7.3, P<0.01) while further analysis of the latter group indicated that adding the predictor of less than 10 inpatients days to age decreased the likelihood of an OLA switch from 51% to 45% (chi(2)=7.0, P<0.01). CONCLUSIONS: ROC analyses have the advantage over other analyses, such as regression techniques, insofar as their "cut-points" are readily interpretable, their sequential use forms an intuitive "decision tree" and allows the potential identification of clinically relevant "subgroups". The software used in this analysis is in the public domain (http://mirecc.stanford.edu). PMID- 14563386 TI - Total length of nerve fibers in prefrontal and global white matter of chronic schizophrenics. AB - It has been suggested that the dysfunction of the prefrontal cortex in schizophrenics is due to dysfunctional connections between the prefrontal cortex and more posterior structures. The present study uses a recent stereological method that allows quantitation of the myelinated nerve fibers in the brain white matter. As especially the prefrontal region is of interest in schizophrenics, the prefrontal white matter was quantitated separately. The total length of nerve fibers in post-mortem brains was estimated from eight male chronic schizophrenics and nine male controls (age-range: 40-81 years). Samples were taken systematically and randomly from both the entire white matter and selectively from the prefrontal white matter. The biopsies were rotated randomly before sectioning to avoid bias due to the anisotropic nature of nerve fibers. The fibers were counted at light microscopic level at about 10,000 x magnification and the fiber diameter of each counted fiber was measured to get the size distribution of the fibers. The schizophrenics had a total of 129,000 km myelinated fibers in the white matter and 25,700 km in the prefrontal white matter, which was non-significantly different from a total of 137,000 km in the entire white matter and 27,600 km in the prefrontal white matter in controls. The size distribution of the fibers in schizophrenics was within normal limits compared to controls. Our results do not show a larger loss of nerve fibers in neither the white matter globally or in the prefrontal white matter of schizophrenics. PMID- 14563387 TI - Decreased platelet peripheral-type benzodiazepine receptors in persistently violent schizophrenia patients. AB - Peripheral-type benzodiazepine receptors (PBR) have been shown to be sensitive to stressful conditions. This study aimed to explore a possible association of platelets PBR binding with aggressive behavior and homicidal history in schizophrenia patients. The authors compared [(3)H] PK 11195 binding to platelet membrane among 11 currently aggressive schizophrenia patients, 15 schizophrenia patients with homicidal history, 14 nonaggressive schizophrenia patients, and 15 healthy volunteers. Subjects were assessed for aggressive behavior, psychopathology, anxiety, anger, and emotional distress using standardized instruments. We found that currently aggressive patients had significantly lower (-30%) platelet PBR density (B(max)), and scored significantly higher on hostility, anxiety, state anger, and emotional distress compared to homicidal and nonaggressive schizophrenia patients and healthy controls. Predominance of positive or negative symptoms, homicidal or suicidal attempt history, emotional distress levels, and conventional or atypical antipsychotic therapy is not associated with the expression of platelet PBR binding sites. Significant negative correlations emerged between PBR density and scores for aggressive behavior, hostility and anxiety. Thus, decreased platelet PBR density in aggressive schizophrenia patients is associated with higher scores for overt aggression, hostility and anxiety, but independent of illness subtype, homicidal and suicidal attempt history, distress level and type of antipsychotic treatment. PMID- 14563389 TI - Guidance for the safety assessment of botanicals and botanical preparations for use in food and food supplements. AB - There is a growing interest by both consumers and industry for the development of food products with 'functional' properties, or health benefits. These products may take the form of dietary supplements or of foods. The health benefits are given by particular ingredients, and in many cases these are derived from botanicals. The variety of plants providing these functions is large, ranging from staple food sources such as cereals, fruits and vegetables, to herbals as used in traditional medicine. The food or ingredient conferring health properties may consist of the plants themselves, extracts thereof, or more purified components. The scientific literature is abundant with articles not only on the beneficial properties, but also on possible adverse health effects of plants and their components. The present report discusses the data required to determine the safe use of these types of ingredients, and provides advice on the development of risk assessment strategies consistent with due diligence under existing food regulations. Product specifications, composition and characterisation of standardised and authentic materials, documented history of use and comparison to existing products (taking into account the effect of industrial processing), description of the intended use and consequent exposure are highlighted as key background information on which to base a risk evaluation. The extent of experimental investigation required, such as in vitro, animal, and/or human studies, depends on the adequacy of this information. A decision tree is presented as an aid to determine the extent of data requirements based on product comparison. The ultimate safety in use depends on the establishment of an adequate safety margin between expected exposure and identified potential hazards. Health hazards may arise from inherent toxicities or contaminants of the plant materials, including the mechanism of the intended beneficial effect. A lower safety margin may therefore be expected than for food ingredients or additives where no physiological effects are intended. In rare cases, post launch monitoring programmes may be envisaged to confirm expected exposures and adequacy of the safety margin. This guidance document was elaborated by an expert group of the Natural Toxin Task Force of the European Branch of the International Life Sciences Institute--ILSI Europe and discussed with a wider audience of scientists at a workshop held on 13-15 May 2002 in Marseille, France. PMID- 14563390 TI - Tin in canned food: a review and understanding of occurrence and effect. AB - Tinplate is light gauge, steel sheet or strip, coated on both sides with commercially pure tin and has been used for well over a hundred years as a robust form of food packaging. Altogether, about 25,000 million food cans are produced and filled in Europe per annum, about 20% of these having plain internal (unlacquered) tin-coated steel bodies. Worldwide, the total for food packaging is approximately 80,000 million cans. Tinplate is also extensively used for the production of beverage cans. Europe produces and fills over 15,000 million tinplate beverage cans per annum all of which are internally lacquered. The use of tinplate for food and beverage packaging, will result in some tin dissolving into the food content, particularly when plain uncoated internal surfaces are used. The Provisional Tolerable Weekly Intake for tin is 14 mg/kg body weight and recommended maximum permissible levels of tin in food are typically 250 mg/kg (200 mg/kg UK) for solid foods and 150 mg/kg for beverages. However, the question arises as to whether evidence exists that such elevated levels of tin in food in any way constitute a risk to human health. This review considers the factors affecting the dissolution of tin, the reported measurements/surveys of actual levels of tin in canned foods and the studies and reports of acute (short term) toxicity relating to the ingestion of elevated levels of tin in food products. Chronic studies are mentioned, but are not covered in detail, since the review is mainly concerned with possible effects from the ingestion of single high doses. From published data, there appears to be a small amount of evidence suggesting that consumption of food or beverages containing tin at concentrations at or below 200 ppm has caused adverse gastrointestinal effects in an unknown but possibly small proportion of those exposed. However, the evidence supporting this assertion is derived from reports of adverse effects which offer data that are limited, incomplete or of uncertain veracity. Clinical studies provide greater confidence regarding the effects of exposure concentration and dose, but few relevant studies have been made. Adverse gastrointestinal effects were observed in limited clinical studies at concentrations of 700 ppm or above, although no adverse gastrointestinal effects were also reported in two studies at higher concentrations. Overall, therefore, the published data do not present a particularly comprehensive profile on the toxic hazard to man of acute exposure to divalent inorganic tin. A food survey suggested that the contents of almost 4% of plain internal tinplate food cans contain over 150 mg/kg of tin and over 2.5 million such cans are consumed every year in the UK alone. Despite this, in the last 25 years, there have been no reports of acute effects attributable to tin contamination in the range 100-200 ppm. These facts strongly suggest that there is little evidence for an association between the consumption of food containing tin at concentrations up to 200 ppm and significant acute adverse gastrointestinal effects. Clearly though, only further clinical studies will generate unequivocal evidence that current legislative limits provide safe levels for adults in the general population. PMID- 14563391 TI - Comparative assessment of gastrointestinal irritant potency in man of tin(II) chloride and tin migrated from packaging. AB - Tin is present in low concentrations in most canned foods and beverages, the highest levels being found in products packaged in unlacquered or partially lacquered tinplate cans. A limited number of case-reports of acute gastrointestinal disorders after consumption of food containing 100-500 mg/kg tin have been reported, but these reports suffer many insufficiencies. Controlled clinical studies on acute effects of tin migrated from packaging suggest a threshold concentration for adverse effects (AEs) of >730 mg/kg. Two separate randomised, single-centre, double-blind, crossover studies, enabling comparison of the tolerability of tin added as tin(II) chloride at concentrations of <0.5, 161, 264 and 529 mg/kg in 250 ml tomato juice in 20 volunteers (Study 1) and tin migrated from packaging at concentrations of <0.5, 201 and 267 mg/kg in 250 ml tomato soup in 24 volunteers (Study 2) were carried out. Distribution studies were conducted to get insight in the acute AEs of low molecular weight (<1000 Da) tin species in the soluble fraction of food products. Results show that the chemical form of tin and not the elemental concentration per se determines the severity of AEs. A clear dose-response relationship was only observed when tin was added as tin(II) chloride in tomato juice. No clinically significant AEs were reported in Study 2 and comparison of the incidence of tin-related AEs showed no difference between the dose levels (including control). Tin species of low molecular weight in supernatant represented 31-32% of total tin in canned tomato soup versus 56-61% in juice freshly spiked with tin(II) chloride. Differences in the incidence of AEs following administration of tomato juice with 161 and 264 mg of tin per kg and tomato soup with 201 and 267 mg of tin per kg likely results from differences in the concentration of low molecular weight tin species and in the nature of tin complexes formed. The results of this work demonstrate that tin levels up to 267 mg/kg in canned food cause no AEs in healthy adults and support the currently proposed tin levels of 200 mg/kg and 250 mg/kg for canned beverages and canned foods, respectively, as safe levels for adults in the general population. PMID- 14563392 TI - Black bean (Phaseolus vulgaris L.) as a protective agent against DNA damage in mice. AB - This study was designed to evaluate the toxicogenetic or protective effect of cooked and dehydrated black beans (Phaseolus vulgaris L.) in bone marrow and peripheral blood cells of exposed mice. The frequency of micronuclei detected using the bone marrow erythrocyte micronucleus test and level of DNA lesions detected by the comet assay were chosen as end-points reflecting mutagenic and genotoxic damage, respectively. Initially, Swiss male mice were fed with a 20% black bean diet in order to detect mutagenic and genotoxic activity. However, no increase in the frequency of bone marrow micronucleated polychromatic erythrocytes (MN PCEs) or DNA lesion in leukocytes was observed. In contrast, received diets containing 1, 10 or 20% of black beans, a clear, but not dose dependent reduction in the frequency of MN PCEs were observed in animals simultaneously treated with cyclophosphamide, an indirect acting mutagen. Similar results were observed in leukocytes by the comet assay. Commercial anthocyanin was also tested in an attempt to identify the bean components responsible for this protective effect. However, instead of being protective, the flavonoid, at the highest dose administered (50 mg/kg bw), induced primary DNA lesion, as detected by the comet assay. These data indicate the importance of food components in preventing genetic damage induced by chemical mutagens, and also reinforce the role of toxicogenetic techniques in protecting human health. PMID- 14563393 TI - Oxidized beta-carotene inhibits gap junction intercellular communication in the human lung adenocarcinoma cell line A549. AB - In addition to its antioxidant activity, beta-carotene (BC) is known to enhance gap junction intercellular communication (GJIC) by up-regulation of connexin 43 (Cx43), an action that may be important in its control of tumor growth. Surprisingly, two clinical trials on supplemental BC suggest that BC may increase lung cancer incidence in smokers. Recently, an animal study indicated that a very high dose of BC (50 mg/kg b.w./day for 5 days) decreases GJIC in rat liver, while a lower dose (5 mg/kg b.w./day) increases GJIC. It is unclear how high-doses of BC inhibit GJIC. In this study, we tested whether oxidized BC (OBC, obtained by heating BC at 60 degrees C in open air for 1 h) may inhibit GJIC. We incubated a human lung cancer cell line (A549) with OBC or BC at 2-10 microM for 5 days. Cell viability (by Trypan-blue assay), GJIC (by scrape-loading dye transfer) and Cx43 expression (by western blotting and immunocytochemical localization) were measured to investigate the effects of OBC and BC on GJIC and the possible mechanisms. The results show that OBC at concentrations lower than 10 microM did not significantly affect cell viability. However, OBC at 5 muM inhibited GJIC, whereas BC at 5 microM markedly increased GJIC. The loss of GJIC in A549 induced by OBC accompanied the aberrant localization and phosphorylation of connexin43 (Cx43). These changes in the expression of Cx43 induced by OBC were similar to those induced by 12-O-tetradecanoylphorbol-13-acetate (TPA), a tumor promoter. Thus, our results suggest that in vivo inhibition of GJIC by a high dose of BC on GJIC is, at least in part, attributable to the effect of OBC. PMID- 14563394 TI - Betaine in sub-acute and sub-chronic rat studies. AB - To evaluate the sub-acute and sub-chronic effect of large doses of betaine, this trimethyl glycine compound was fed to rats. Initial studies at BIBRA in the UK evaluated intakes of 0, 1, 2, and 5% betaine added to a maintenance chow designed for use in toxicology studies. Male and female Sprague-Dawley rats were followed for up to 90 days. No toxicity occurred, but at higher betaine intakes several serum chemistries were altered slightly, the MCV, MCH, and MCHC of red cells were reduced, and hepatocytes developed fatty droplets in direct proportion to betaine intake. Females were more affected than males. In a second study to assess reversibility in females, betaine effects were induced for 28 days, followed by a 28 day betaine-free period. All perturbations, except the reduced MCV and MCH, were reversed. As a follow up to BIBRA investigations, both 28 and 90 day feeding trials were conducted at Brandeis University using a rat chow with higher levels of energy, protein, and fat, with betaine added at 0, 0.5, 0.75, 1.0 and 5.0% of the diet. A similar broad range of clinical chemistries and physiological parameters were monitored, and hepatic lipid droplets were investigated in more detail. Liver lipid was actually reduced by betaine, and no significant adverse effects of clinical importance resulted from any dose. However, the MCV was again reduced at 5% betaine in the 28 day study. By 90 days all parameters were normal and comparable to controls. Based on these collective data, it was concluded that even at these high doses, betaine is nontoxic. Differences observed between the BIBRA and Brandeis studies were attributed to differences in the dietary formulations. Significant betainexdietxgrowth interactions were thought to reflect primary disparities in protein and energy concentrations, more than the addition of betaine per se. PMID- 14563395 TI - An in vitro model system to predict the bioaccessibility of heterocyclic amines from a cooked meat matrix. AB - To understand the impact of variation in digestion parameters on the release of heterocyclic amines naturally formed during cooking, we developed and characterized a model system to assess the effect of amylase, pepsin, and pancreatin on digestion of well-done chicken. The amounts of MeIQx, DiMeIQx, IFP, and PhIP in the liquid portion of the digestate were compared to levels in the undigested meat to determine the percentage released (accessible fraction). Incubating the meat with amylase and pepsin did not change the accessibility of HAs when compared to incubation with water alone. In contrast, increasing amounts of pancreatin increased the accessibility up to 6.4-fold. Comparing the amounts of the HAs in the liquid to the solid fraction showed that there was more MeIQx, DiMeIQx, and IFP in the liquid fraction. In contrast, PhIP was equally divided between the solid and liquid fractions. For all four compounds, increasing the doneness of the meat decreased the amount of the compound accessible from the meat matrix. Our data suggest that bioaccessability of HAs may vary according to the polarity of the individual HAs and also may depend upon the doneness of the meat. These results may have important ramifications for human feeding studies, which assume that the total amount of each HA in the meat matrix is equally bioavailable. PMID- 14563396 TI - Effects of environmental estrogenic compounds on growth of a transplanted estrogen responsive pituitary tumor cell line in rats. AB - The modifying effects of three endocrine disrupting chemicals on the promotion phase of estrogen dependent tumor development were investigated with a transplantable rat pituitary cell line, MtT/E-2. The EDCs examined were genistein (Gen), a phytoestrogen, p-nonylphenol (NP), a surfactant and atrazine (Atz), a herbicide. Since potential exposure is through food and drinking water, oral administration was examined in the present study. Gen and NP stimulated in vitro MtT/E-2 growth at concentrations of 10(-8) and 10(-6) M, respectively, while Atz did not show any effects. The estrogenic activity was further confirmed by measuring transcription of an ERE-luciferase reporter transiently transfected in the cells. When MtT/E-2 cells are inoculated into ovariectomized female F344 rats, estrogen dependent tumors develop providing a simple and sensitive model to examine modulation effects of estrogenic compounds in the promotion phase. Ovariectomized F344 rats implanted with MtT/E-2 were fed diet containing NP or Gen at 25 and 250 mg/kg, or Atz at 5, 50 and 500 mg/kg. NP and Atz did not exert any modifying effects on pituitary tumor development, while Gen at 250 mg/kg exhibited a promotion influence. These results indicate that Gen might facilitate the estrogen responsive tumor development if only the promotion phase is concerned, while NP and Atz at doses used in the present study were without effect. PMID- 14563397 TI - RAPD markers system as a useful tool for rapid identification of the origin of lizard contaminants in food. AB - In food poisoning, detection of the nature of causative agent is important for management of trauma and forensic investigation. Most of the methods in clinical toxicology are developed for detection of toxins and poisons. A RAPD-based method has been described for detection of species of animal from its morphologically unrecognizable fragments, recovered from food substances, consumption of which caused even death. Pre-mixed RAPD reaction beads and six RAPD primers were used in PCR analysis. Among six RAPD primers used, any one of them was sufficient in resolving this practical forensic situation. But to enhance the probability values for matching in the present study of fixing identity of an animal, six set of market available RAPD primers were used. This is the first report of a forensic application of RAPD DNA typing in identification of charred skeleton remnants of Lizard sp. in food material. Furthermore unique amplicons were generated for different reptilian species, which can be used as species specific markers for species identification in forensic situation, however no variations among individuals of same species were observed. PMID- 14563398 TI - Isolation and structure elucidation of the major individual polyphenols in carob fibre. AB - Although it is already known that carob fibre contains several classes of polyphenolic substances, a comprehensive analysis of these has not been conducted to date. Therefore, the major polyphenolic compounds were extracted with organic solvents, and, following fractionation by normal-phase column chromatography on silicic acid, their structures were elucidated by liquid-chromatography electrospray-ionisation mass spectrometry (LC-ESI), nano-electrospray-ionisation mass spectrometry (ESI-MS), and gas-chromatography mass spectrometry (GC-MS). In addition, complete 1H and 13C NMR assignments were obtained for the isolated gallotannins 1,6-di-, 1,2,6-tri- and 1,2,3,6-tetra-O-galloyl-beta-D-glucose. Carob fibre was found to contain a rich variety of phenolic antioxidants. A total of 24 polyphenol compounds were identified with a yield of 3.94 g/kg (dry weight). The profile was dominated by gallic acid in various forms: free gallic acid (42% of polyphenols by weight), gallotannins (29%), and methyl gallate (1%), while simple phenols, mainly cinnamic acid, made up about 2% of the total. Flavonoids represented 26% of the polyphenols, and the major components were identified as the glycosides myricetin- and quercetin-3-O-alpha-L-rhamnoside (ca. 9% and 10%, respectively). These data indicate that carob fibre is rich in both amount and variety of phenolic antioxidant substances, and its inclusion in the diet may have chemopreventive properties. PMID- 14563399 TI - Effects of high fat fish oil and high fat corn oil diets on initiation of AOM induced colonic aberrant crypt foci in male F344 rats. AB - Modulating effects of high fat fish oil (HFFO) and high fat corn oil (HFCO) diets on azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) were studied in male F344 rats following 8 weeks of dietary treatment. The incidence of AOM induced ACF was significantly lower in the proximal colon of rats fed the HFFO diets compared with rats fed the HFCO diets. No differential effects were found on enzyme activities that are involved in metabolic activation and detoxification of AOM. Activities of hepatic P450 IAI and P450 IIBI and hepatic and feacal levels of lipid peroxidation were increased by feeding the HFFO diet. Hepatic GST activity and plasma levels of PGE(2) were significantly lower in rats fed the HFFO diets compared with those fed the HFCO diets. These observations demonstrate that HFFO diets with high levels of n-3 PUFAs are also protective against preneoplastic lesions in the early stages of chemically induced colon carcinogenesis. It seems unlikely from our results that the inhibitory effect of a HFFO diet can be attributed to an altered metabolic activation and detoxification of AOM. Other mechanisms such as oxidative stress or reduction of PGE(2) levels may play an important role in the anticarcinogenic effects of n-3 PUFAs. PMID- 14563400 TI - A subchronic 90-day oral rat toxicity study and in vitro genotoxicity studies with a conjugated linoleic acid product. AB - Conjugated linoleic acid (CLA) is the term given to a group of positional and geometric isomers of the essential fatty acid linoleic acid. CLA is found naturally in foods such as dairy and meat products. CLA is reported to have a number of beneficial effects including anticarcinogenic activity. However, safety data are limited. Clarinol G80 is a commercial preparation containing equal amounts of the 9cis,11trans and 10trans,12cis CLA isomers in the form of glycerides. In order to support the safety-in-use of Clarinol G80 as an ingredient in food, the preparation was tested in two in vitro mutagenicity assays, an Ames test and an in vitro cytogenetics assay, and a 90-day repeat-dose oral toxicity rat study. Clarinol G80 was non-mutagenic in both in vitro assays. In the 90-day study, Clarinol G80 produced hepatocellular hypertrophy in female rats at the highest dose level (15% w/w). This effect was an adaptive effect in response to feeding high levels of Clarinol G80 in the diet and was reversible upon withdrawal of test material. An increase in plasma insulin levels was also observed female rats fed 15% w/w Clarinol G80 but there was no effect on plasma glucose levels. A No Observed Adverse Effect Level of 2433 mg/kg bw/day for male and 2728 mg/kg bw/day female rats was identified in the study. PMID- 14563401 TI - Studies on the metabolism of the thiofurans furfuryl mercaptan and 2-methyl-3 furanthiol in rat liver. AB - The metabolism of two thiofurans, namely furfuryl mercaptan (FM) and 2-methyl-3 furanthiol (MTF), to their corresponding methyl sulphide and methyl sulphoxide derivatives has been studied in male Sprague-Dawley rat hepatocytes and liver microsomes. Rat hepatocytes converted FM to furfuryl methyl sulphoxide (FMSO) and MTF to 2-methyl-3-(methylthio)furan sulphoxide (MMFSO). Liver microsomes catalysed the NADPH-dependent metabolism of furfuryl methyl sulphide (FMS) to FMSO and 2-methyl-3-(methylthio)furan sulphide (MMFS) to MMFSO. FMS and MMFS metabolism to their thiofuran methyl sulphoxide derivatives was induced by the treatment of rats with Aroclor 1254 and inhibited in liver microsomes treated with 1-aminobenzotriazole. The NADPH-dependent metabolism of FM to FMSO and MTF to MMFSO in liver microsomes was observed in the presence of S adenosylmethionine. In summary, both thiofurans can be metabolised in rat liver to their thiofuran methyl sulphide derivatives which can be subsequently S oxidised to form thiofuran methyl sulphoxides. FM and MTF appear to be substrates for rat hepatic microsomal thiol methyltransferase and the S-oxidation of FMS and MMFS appears to be primarily catalysed by cytochrome P450 forms. PMID- 14563402 TI - Toxicological evaluation of propane expanded tobacco. AB - A tiered testing strategy has been developed to evaluate the potential for tobacco processes, ingredients, and other technological developments to increase or decrease the biological activity resulting from burning tobacco. The strategy is based on comparative chemical and biological testing. Propane expanded tobacco is an example of a processed tobacco used in the modern manufacture of cigarettes. Test cigarettes containing propane expanded tobacco were compared to control cigarettes containing tobacco expanded with a traditional expansion agent (Freon-11). The toxicological evaluation included chemistry studies using mainstream cigarette smoke (determination of selected constituent yields), in vitro studies using cigarette smoke condensate (Ames study in Salmonella typhimurium and sister chromatid exchange study in Chinese hamster ovary cells) and in vivo studies (13-week inhalation study of mainstream cigarette smoke in Sprague-Dawley rats and 30-week dermal tumor promotion study of cigarette smoke condensate in SENCAR mice). Although statistically significant differences in several smoke constituents were observed, most constituents from cigarettes containing 100% propane expanded tobacco were within market survey ranges. Furthermore, biological tests indicated that the cigarettes containing propane or Freon-11 expanded tobacco were not significantly different. PMID- 14563403 TI - The in vivo assessment of safety and gastrointestinal survival of an orally administered novel probiotic, Propionibacterium jensenii 702, in a male Wistar rat model. AB - This study aimed to evaluate in vivo gastrointestinal survival and safety of orally administered probiotic bacterium, Propionibacterium jensenii 702, using a male Wistar rat model. A high dose of 10(10) cfu/rat/day of P. jensenii 702 was fed to each rat for 81 days. The repeated dose toxicity and translocation of P. jensenii 702 into rat tissues were evaluated, along with the rat faecal beta glucuronidase activities and dairy propionibacteria counts. Results showed that P. jensenii 702 had no adverse effect on general health status, body weight gain, visceral organs and faecal beta-glucuronidase activities. No viable cells of P. jensenii 702 were recovered from blood and tissue samples (mesenteric lymph nodes, liver and spleen) of rats, and no treatment-associated illness or death was observed. Faecal dairy propionibacteria counts reached 10(8) cfu/g after 36 days treatment and remained between 10(8)-10(9) cfu/g till the end of 81 days treatment. The results indicate that P. jensenii 702 was able to survive the in vivo gastrointestinal tract transit of rats, with no adverse affects on the animals. However, further human clinical trials are required before strain P. jensenii 702 could be incorporated into food for human consumption as probiotics. PMID- 14563405 TI - Classification of contact allergens according to potency: proposals. AB - It is clear that contact allergens vary substantially with regard to the relative potency with which they are able to induce skin sensitisation. Considerations of potency will in the future become a significant factor in the classification of skin sensitising chemicals. It is therefore appropriate to establish what is known of potency and thresholds in the induction of skin sensitisation and the elicitation of allergic contact dermatitis, and to identify approaches that might be available for assessment of relative potency for the purposes of categorising chemical allergens. This paper was prepared by an ECETOC (European Centre for Ecotoxicology and Toxicology) Task Force that had the objective of recommending approaches for the measurement of potency and definition of thresholds for both the induction and elicitation of contact sensitisation. The deliberations recorded here build upon recommendations made previously by an ECETOC Task Force that considered the conduct of standard skin sensitisation test methods for the purposes of hazard identification and risk assessment (ECETOC, Monograph No. 29, Brussels, 2000). The emphasis in this present paper is also on standard and accepted methods for the assessment of skin sensitisation, and for which OECD guidelines are available: the local lymph node assay (LLNA), the guinea pig maximisation test and the occluded patch test of Buehler. For various reasons, discussed in detail herein, attention focused primarily upon consideration of categorisation of chemical allergens and the identification of thresholds with respect to the induction of skin sensitisation, rather than the elicitation of allergic contact dermatitis. It is concluded that although the LLNA is the method of choice for the determination of skin sensitisation potency for the purposes of categorisation, if data are already available from appropriate guinea pig tests then their judicious interpretation may provide information of value in determinations of potency and categorisation. Included here are detailed and specific recommendations for how best the results of the three test methods considered can be used for the categorisation of chemical allergens as a function of skin sensitisation potency. PMID- 14563404 TI - Protein deficiency and muscle damage in carbon tetrachloride induced liver cirrhosis. AB - Protein undernutrition, alterations of hormones such as IGF-1, testosterone and cortisol, and increased lipid peroxidation-which may be related with deranged metabolism of some elements such as iron (Fe), zinc (Zn), manganese (Mn), selenium (Se) or copper (Cu)-may contribute to muscle damage in non alcoholic cirrhosis. Here, we analyse the effect of protein deficiency on muscle Cu, Fe, Zn, Mn and Se in carbon-tetrachloride (CCl(4)) induced liver cirrhosis. We also study the association between protein undernutrition and these trace elements with the activity of glutathione peroxidase (GPX), superoxide dismutase (SOD) and lipid peroxidation products, and how all these are related with muscle morphological changes in 40 male adult Sprague-Dawley rats. Liver cirrhosis was induced by intraperitoneal injection of CCl(4) to 10 rats fed a 2% protein diet, and to another 10 fed a 18% protein control diet. Two further groups included rats without cirrhosis fed the 2% protein and the 18% protein diets. After sacrifice (6 weeks later), we found type IIa fibre atrophy in the cirrhotic animals, especially in the low-protein fed ones and this was due to protein deficiency. Muscle Fe increased in low protein fed cirrhotic rats. No relationship was found between muscle changes and any of the hormones, enzymes and trace elements analysed, or with liver fibrosis. These results suggest that muscle atrophy observed in CCl(4)-induced cirrhosis is related with protein deficiency, but not with cirrhosis itself. PMID- 14563406 TI - Lack of modifying effects of atrazine and/or tamoxifen on thyroid carcinogenesis in rats pretreated with N-bis(2-hydroxypropyl)nitrosamine (DHPN). AB - The modifying effects of atrazine, and/or tamoxifen, on thyroid carcinogenesis were investigated in a rat two-stage carcinogenesis model following N-bis(2 hydroxypropyl)nitrosamine (DHPN) initiation. Five-week-old male F344 rats were given a single subcutaneous injection of DHPN (2800 mg/kg, body weight) or vehicle alone. Starting 1 week later, the animals were fed a diet supplemented with 0, 5, 50 or 500 ppm of atrazine, 500 ppm atrazine plus 5 ppm tamoxifen, or 5 ppm tamoxifen in the DHPN-treated groups, and 0 or 500 ppm of atrazine in the DHPN-untreated groups for 24 weeks. At autopsy major organs, including the thyroid, pituitary, liver, kidney, testis, epididymis, and brain, were collected and histopathologically examined. Body weights were significantly (P<0.05) decreased by the high doses of atrazine or tamoxifen, the effect being enhanced in combination. Relative thyroid weights were significantly increased (P<0.05) only in the tamoxifen-treated group and pituitary weights were elevated with 500 ppm atrazine plus tamoxifen (P<0.05). Relative liver weights were increased by the high dose of atrazine. However, the atrazine and/or tamoxifen treatments did not induce significant histopathological changes in the major organs, including the thyroid, nor cause significant changes in serum TSH levels. These results suggest that neither atrazine nor tamoxifen may promote thyroid carcinogenesis, alone as well as in combination. PMID- 14563407 TI - Evaluation of urinary N-acetyl cysteinyl allyl isothiocyanate as a biomarker for intake and bioactivity of Brussels sprouts. AB - Isothiocyanates (ITC), glucosinolate hydrolysis products from Brussels sprouts (BS) and other cruciferous vegetables, are considered to protect the body from cancer by induction of detoxification enzymes such as quinone reductase (QR). Urinary N-acetyl-cysteine (NAC) conjugates of ITC have been proposed as biomarkers of crucifer intake. Here we asked if dietary intake and induction of detoxification enzymes are dose-related to urinary NAC conjugate appearance. Male F344 rats (4/group) received an AIN 76B-40 diet containing 0, 10 or 20% freeze dried BS for 6 days. A human subject ingested 500 g BS. Urinary AITC-NAC was identified in human and rat urine. Ten and 20% BS diets caused a 1.4- and 2.3 fold induction of QR in the pancreas, a 1.5- and 2.5-fold induction in liver and a 3.1- and 3.6-fold induction in colonic epithelium, respectively. Liver and pancreatic QR induction was dose-related, whereas induction of QR in colon was less different between the two doses. Excretion of the conjugate was dose-related only on day 1, and unrelated to dose after day 2. These results suggest that urinary NAC-AITC is a qualitative biomarker for ingestion and bioactivity of BS, but that it may not be dose-related when rats are fed continuously for 2 or more days. PMID- 14563408 TI - Triacsin C inhibits the formation of 1H NMR-visible mobile lipids and lipid bodies in HuT 78 apoptotic cells. AB - Nuclear magnetic resonance-visible mobile lipids (ML) have been reported to accumulate during cell apoptosis in vitro and in vivo. The biogenesis, biochemical nature and structure of these lipids are still under debate. In this study, a human lymphoblastoid cell line, HuT 78, was induced to apoptosis by exposure to anti-Fas monoclonal antibodies (alpha-Fas mAb) followed by incubation for different time intervals (1-24 h, hypodiploid cell fraction, H, varying from 1% to over 60%) either in the presence or in the absence of 5.0 microM Triacsin C (TRC), specific inhibitor of long-chain acyl-CoA synthetase (ACS). The increase of ML in apoptotic cells correlated linearly with H and was associated with: (a) accumulation of intracellular lipid bodies, detected by confocal laser scanning microscopy in lipophilic dye-stained cells; (b) increases, detected by thin-layer chromatography in total lipid extracts, in the relative abundance of triacylglycerides (TAG) and cholesteryl esters (CE), with corresponding decreases of phospholipids (PL). TRC completely abolished both ML and lipid body formation in anti-Fas-treated apoptotic cells, with concomitant reversion of TAG, CE and PL to control levels, but did not alter cell viability nor did it inhibit apoptosis. ML signals detected during anti-Fas-induced apoptosis therefore appear to originate from neutral lipids assembled in intracellular lipid bodies, synthesised from cellular acyl-CoA pools. PMID- 14563409 TI - Genomic structure and genitourinary expression of mouse cytosolic prostaglandin E(2) synthase gene. AB - Prostaglandin E(2) (PGE(2)) plays an important role in genitourinary function. Multiple enzymes are involved in its biosynthesis. Here we report the genomic structure and tissue-selective expression of cytosolic PGE(2) synthase (cPGES) in genitourinary tissues. Full-length mouse cPGES cDNA was cloned by reverse transcript-polymerase chain reaction (RT-PCR) and 5'- and 3'-rapid amplification of cDNA ends (RACE). Analysis of a cPGES cDNA with partially sequenced cPGES genomic clones and bioinformatic databases demonstrates that the murine cPGES gene spans approximately 22 kb and consists of eight exons. The cPGES gene promoter is GC-rich and contains many SP1 sites but lacks an obvious TATA box motif. RNase protection assay revealed constitutive expression of cPGES was greatest in the testis with lower levels in the ovary, kidney, bladder and uterus. In situ hybridization studies demonstrated that cPGES mRNA was most highly expressed in the epithelial cells of seminiferous tubules in the testis. In the female reproductive tissues, cPGES was mainly localized in ovarian primary and secondary follicles and oviductal epithelial cells with less expression in uterine endometrium. In the kidney cPGES expression was diffusely expressed. In urinary bladder, cPGES expression was restricted to the transitional epithelial cells. This expression pattern is consistent with an important role for cPGES mediated PGE(2) in urogenital tissue function. PMID- 14563410 TI - Oxidation affects the flow-induced aggregation of low density lipoprotein and its inhibition by albumin. AB - We investigated whether oxidation alters the self-aggregation of low density lipoprotein (LDL) and the inhibition of such aggregation by albumin. Incubation with copper for different durations produced mildly, moderately, and highly oxidised LDL (having, respectively, ca. 60, 300 and 160 nM lipid hydroperoxides/mg protein, and electrophoretic mobilities 1.2, 2.6 and 4.4 times that of native LDL). The rate of flow-induced aggregation was the same for native, mildly oxidised and moderately oxidised LDL, but decreased for highly oxidised LDL. The inhibitory effect of albumin (40 mg/ml) on aggregation was reduced by mild oxidation and further reduced by moderate or severe oxidation. The net result of the two effects was that in the presence of albumin, moderately oxidised LDL had the highest rate of aggregation and native the lowest. The reduction in the anti-aggregatory effect of albumin provides a new mechanism by which LDL oxidation might enhance net aggregation in vivo. PMID- 14563411 TI - Ceramide induces translocation of protein kinase C-alpha to the Golgi compartment of human embryonic kidney cells by interacting with the C2 domain. AB - Ceramide is a lipid second messenger produced by sphingolipid metabolism in cells exposed to a limited number of agonists and in turn triggers important cell responses including protein kinase C (PKC)-alpha activation. Using a fusion protein comprising bovine PKCalpha and the green fluorescent protein (GFP), we transfected human embryonic kidney (HEK) cells and investigated to which subcellular compartment ceramide triggers PKCalpha redistribution. Stimulation of HEK cells with exogenous C16-ceramide or bacterial sphingomyelinase (bSMase), which leads to increased endogenous ceramide formation, evokes a translocation of PKCalpha to the Golgi compartment. By using deletion mutants of PKCalpha lacking distinct domains in the regulatory region, it is shown that the Ca(2+)-dependent lipid binding C2 domain, but not one of the C1 domains is essentially required for the ceramide-triggered translocation of PKCalpha to the Golgi complex. In contrast, the C2 domain is not required for phorbol ester (TPA) binding and translocation of PKCalpha to the plasma membrane. In addition, evidence is provided that TPA requires only one of the two C1 subdomains to trigger translocation to the plasma membrane.In summary, our data provide evidence that ceramide either directly or indirectly interacts with the Ca(2+)-dependent lipid binding C2 domain of PKCalpha and thereby induces translocation of the enzyme to the Golgi compartment. PMID- 14563412 TI - Saturated free fatty acid release and intracellular ceramide generation during apoptosis induction are closely related processes. AB - Apoptosis induced by cells from the immune system is frequently associated with an increase in the ceramide content of target cells, due to the activation of sphingomyelinases (SMase). Some studies have also reported the release of saturated and monounsaturated free fatty acids (FFA) from apoptotic cells. However, the possible relationship between these lipid biochemistry events has not been characterized. We have analysed for the first time the release of FFA triggered by tumor necrosis factor-alpha (TNF-alpha), Fas/CD95 or the perforin/granzyme system of cytotoxic T lymphocytes (CTL) and their relationship to intracellular ceramide generation. TNF-alpha- and Fas-induced apoptosis are associated with both intracellular ceramide generation from sphingomyelin (SM) and release of palmitic-derived FFA, with similar kinetics. Intracellular SMase activation and FFA release from target cells during Fas-induced apoptosis are much more rapid and efficient if Fas-based cytotoxicity is exerted by alloantigenic CTL. In the case of perforin/granzyme-based cytotoxicity exerted by CTL, intracellular ceramide generation and FFA release from target cells seem to depend on the type of lysis induction used. Importantly, the correlation between intracellular SMase activation and the release of palmitic acid-derived FFA from target cells has been observed in all types of cytotoxicity assayed. In addition, exogenous natural ceramide induces the rapid release of the same FFA, well before any apoptotic sign is detected, and FFA release during Fas-induced apoptosis is inhibited in SM-depleted cells by chronic fumonisin-B(1) treatment. These results demonstrate a novel connection between the release of palmitic acid-derived FFA and intracellular ceramide accumulation during apoptosis induction. PMID- 14563413 TI - Synthetic capacity of Arabidopsis phosphatidylinositol synthase 1 expressed in Escherichia coli. AB - Phosphatidylinositol (PtdIns) synthase 1 from the plant Arabidopsis thaliana has been expressed in Escherichia coli in order to study the synthetic capacities of the enzyme. Analysis of the total fatty acid content of the bacteria shows that PtdIns synthase activity does not have a profound effect on the proportions of the different fatty acids produced, even if the presence of an extra acidic phospholipid leads to a global reduction of the lipid content. A closer analysis carried out on individual phospholipids reveals a global fatty acid composition almost unchanged in the two major bacterial lipids phosphatidylethanolamine (PtdEtn) and phosphatidylglycerol (PtdGro). Phosphatidylinositol has a very unusual composition that shows the ability of the plant enzyme to use CDP diacylglycerol molecular species absent from plants. We identified the various PtdIns molecular species. They represent a pool of the major molecular species of PtdEtn and PtdGro. These results, together with the determination of the apparent affinity constants of AtPIS1 for myo-inositol and CDP-diacylglycerol, allow us to discuss some of the constraints of PtdIns synthesis in plants in terms of specificity, which will depend on the subcellular localization of the protein. PMID- 14563415 TI - [Fever-shivers reaction and standard platelet concentrates transfusion: a prospective study]. AB - Fever-shivers reaction (FSR) is the most frequent transfusion immediate incident related to platelet transfusions. The aim of our prospective study was to assess the frequency of the different immediate incidents, especially the frequency and the causes of the FSR, observed during the transfusion of standard platelet concentrates (SPC). For each FSR, analysis of causes included: a bacterial culture of the implicated SPC, a blood culture and HLA antibody screening (lymphocytotoxicity assay) among the patients. In the study period, 34 patients were followed during 74 transfusions. Ten immediate incidents were noted; FSR: N = 8, erythema-urticaria: N = 1 and nausea-vomit: N = 1. The FSR was observed in 6 patients who received 56 SPC. Analysis of causes of this reaction revealed that: HLA antibodies were present in one patient; bacterial contamination was not found neither among the patients nor in the implicated SPC, and the risk of the FSR occurrence rose with increased storage time of the SPC transfused. Indeed, a significant difference was noted between the mean age of the SPC implicated in the FSR and the mean age of those not implicated (P = 0,0028). In conclusion, the FSR is a frequent incident observed during SPC transfusions. In the majority of cases, the cause of this reaction was not identified. Further studies will be necessary to better understand the physiological mechanisms of the FSR. PMID- 14563416 TI - In vitro quality control of red blood cell concentrates outdated in clinical practice. AB - The properties of red blood cell (RBC) concentrates stored in different additive solutions have been previously examined under laboratory conditions at the end of shelf-life. However, whether these data are representative for RBC units used in clinical practice has not been shown. Therefore, we examined 164 RBC units from six manufacturers outdated after clinical usage in a hospital-based transfusion service for cellular content, hemolysis, adenosin triphosphate, 2,3-DPG, pH, oxygen saturation and levels of beta-thromboglobulin and proinflammatory cytokines interleukin (IL) 1beta (IL-1), IL-6, IL-8 and tumor necrosis factor alpha (TNFalpha). Results were correlated with the number of interruptions of recommended storage conditions and with different manufacturers. TNFalpha and IL 8 levels in the supernatant of RBC concentrates showed a weak correlation with the number of interruptions of recommended storage conditions (TNFalpha: r = 0.25, P < 0.01; IL-8: r = 0.20, P < 0.01) for the whole series. We detected no significant correlation between hemolysis and interruptions of recommended storage conditions or any of the remaining studied parameters. However, we found significant differences between RBC concentrates supplied by different manufacturers with respect to cellular content and most of the studied parameters. RBC concentrates containing SAG-M from one single manufacturer had higher in vitro hemolysis at the end of shelf-life compared to all other manufacturers (P < 0.05). We conclude from our data that interruptions of optimal conditions for storage of red cell components during cross-match testing and transport in our setting play a minor role for in vitro properties of RBC units at the end of shelf-life. The influence of processes of production, storage and/or transport until entry of RBC units into our blood component depot seems to be much more important for final product quality at the end of shelf-life than subsequent events. PMID- 14563417 TI - [A quantitative determination of IgG anti-D subclasses by Elisa in hemolytic disease of the newborn]. AB - The quantification of IgG anti-D subclasses is one of the most important parameters considered in the assessment of the severity of hemolytic disease of the newborn. Traditionally IgG subclassing is performed using qualitative haemagglutination methods, difficult to interpret. A quantitative enzyme-linked immunosorbent assay (Elisa) was implemented for measuring IgG anti-D subclasses in 20 sera collected from 14 RhD-immunized pregnant women. All 4 IgG subclasses were detected in the 20 sera tested. The mean proportion of IgG1 was 52.8%. The mean proportion of IgG3 was 30.7%. The mean proportions of IgG2 and IgG4 were 14.5 and 1.9% respectively. A good correlation between the sum of IgG subclasses and the severity of HDN was found. Severe HDN occurred when both IgG1 and IgG3 were present. IgG1 anti-D was the predominant subclass in 4 of the 8 severe cases. PMID- 14563418 TI - [Is GB virus C alias "hepatitis" G virus involved in human pathology?]. AB - GB virus-C alias "hepatitis" virus G was discovered in 1995 as a putative causative virus of non A-E hepatitis. It is a very common virus found in 1 to 5% of eligible blood donors in developed countries. Numerous studies over seven years led to the exclusion of its role as a significant etiological agent of hepatitis. Its in vivo replication site is still unknown. Its direct involvement in the induction of significant hepatic or extra-hepatic diseases could not be demonstrated. However, coinfections with other viruses may contribute to changes in the evolution of both liver disease (negatively) and HIV/AIDS (favourably). Today, no country has decided to screen GBV-C in blood donors. However, more studies are necessary before the absence of influence of GBV-C infection on human health in the context of other viral infections could be confirmed definitely. This article is a review of the literature on a possible involvement of GBV-C in pathologies whether associated or not to other infections. PMID- 14563419 TI - [Production of a new monoclonal anti-AB reagent]. AB - Techniques of monoclonal antibodies production have well progressed in the developed countries. But, they are still absent in the third world. In this report, the murine hybridoma technology was performed to produce anti-ABH monoclonal antibodies. The anti-AB specificity of several hybridomas was determined by agglutination techniques using phenotyped red cells panels. An anti AB monoclonal antibody was selected and compared to other reagents for its use as a blood grouping reagent. Its characteristics were in accordance with the international standards. Hybridoma establishment to produce a good monoclonal reagent is henceforth possible in Morocco. PMID- 14563420 TI - [How the information system can contribute to the implementation of a risk management program in a hospital?]. AB - Nowadays, information system is recognised as one of the key points of the management strategy. An information system is regarded conceptualised as a mean to link 3 aspects of a firm (structure, organisation rules and staff). Its design and implementation have to meet the objectives of medical and economical evaluation, especially risk management objectives. In order to identify, analyse, reduce and prevent the occurrence of adverse events, and also to measure the efficacy and efficiency of the production of care services, the design of information systems should be based on a process analysis in order to describe and classify all the working practices within the hospital. According to various methodologies (usually top-down analysis), each process can be divided into activities. Each activity (especially each care activity) can be described according to its potential risks and expected results. For care professionals performing a task, the access to official or internal guidelines and the adverse events reporting forms has also to be defined. Putting together all the elements of such a process analysis will contribute to integrate, into daily practice, the management of risks, supported by the information system. PMID- 14563421 TI - Usefulness of serum KL-6 for early diagnosis of idiopathic pulmonary fibrosis in patients with hepatitis C virus. AB - The aim of this study was to evaluate when the serum level of KL-6, a sensitive marker for idiopathic pulmonary fibrosis (IPF), rise prior to clinical onset of IPF. Eight hepatitis C virus (HCV)-positive patients with IPF were enrolled in this trial. The serum samples of these eight patients were stored -80 degrees C every 1-3 month during a follow-up period and enzyme-linked immunosorbent assay (ELISA) for KL-6 was done at the same time. Diagnosis of IPF was based on computed tomography and/or histology. Diagnosis of clinical onset of IPF was based on presence of dyspnea, dry cough, and audible fine crackles. At 1 year before clinical onset of IPF, the sensitivities of serum marker for IPF were 75% (6/8) for KL-6, 25% (2/8) for each of lactic acid dehydrogenase (LDH) and C reactive protein (CRP). At 2 years before clinical onset of IPF, the sensitivities of the same serum markers were 62.5% (5/8), 12.5% (1/8) and 0% (0/8), respectively. The sensitivity of KL-6 at 1 and 2 year before clinical onset of IPF was significantly higher compared with LDH and CRP. Our results indicate that many future patients with IPF may have high levels of serum KL-6 at 1 or 2 years before clinical onset of IPF, suggesting that changes in serum KL-6 level can provide useful information for the early diagnosis of IPF in patients with HCV. PMID- 14563422 TI - Clinical significance of TT virus (TTV) infection in chronic hepatitis C patients with high dose interferon-alpha therapy in Taiwan: re-evaluated by using new set of TTV primers. AB - BACKGROUND: The clinical significance of TT virus (TTV) coinfection in chronic hepatitis C (CHC) patients and influence of TTV viremia on hepatitis C virus (HCV) response to high dose interferon-alpha therapy in Taiwan were investigated. MATERIALS AND METHODS: Total 102 HCV RNA-positive CHC patients were enrolled. TTV DNA (using polymerase chain reaction primers derived from 5' non-coding region and open reading frame 2), alanine aminotransferase (ALT), GB virus-C/hepatitis G virus (GBV-C/HGV) RNA, anti-E2 antibody, genotype and RNA levels of HCV were tested. RESULTS: The prevalence of TTV DNA was 51.0%. The mean age of TTV viremic CHC patients was significant higher than non-viremic ones (P<0.05). HCV sustained viral response (SVR) was achieved in 42 (41.2%) patients. Based on multivariate regression analyses, SVR were significantly associated with low pretreatment HCV RNA levels, HCV genotype non-1b and high pretreatment levels of ALT but not TTV viremia. CONCLUSIONS: TTV viremia is highly prevalent among Taiwanese CHC patients and related to increased ages. Neither severity of liver disease nor replication and genotype distribution of HCV was affected by concurrent TTV infection. With high HCV SVR rate associated with pretreatment HCV RNA and ALT levels and HCV genotype, TTV viremia did not influence the HCV response. PMID- 14563423 TI - Differential induction of serum interleukin-6 and -12 by interferon-alpha and beta administration in chronic hepatitis C patients. AB - Interferon (IFN) is widely used to manage chronic hepatitis C virus (HCV) infections. It contributes to the production of various cytokines, and alters cytokine interactions in vivo. Among them, interleukin (IL)-12, is thought to shift the immune system to Th-1 dominance and to, therefore, have a beneficial effect on eradication of HCV. On the other hand, IL-6 seems to be related to the acute inflammatory phenomenon, such as fever accompanied with IFN therapy. We studied the time courses of the serum IL-12 and IL-6 levels of patients with HCV infections treated with either IFN-alpha or IFN-beta. On the first day, both IFN alpha and IFN-beta significantly induced serum IL-6, but when the administration was repeated, the serum IL-6 levels gradually decreased in both the IFN-alpha and the IFN-beta groups. The serum IL-6 levels correlated well with the daily peak body temperature of the patients, which also went down with repeated IFN administration. Serum IL-12 production was induced by IFN-alpha not by IFN-beta. The induced IL-12 decreased to baseline by repeating IFN-alpha administration. These results suggested that IFN-alpha and IFN-beta induce cytokine production in a different manner, and that their effect on cytokine interactions is short-term and reduced by repeated administration. Therefore, the induction of serum IL-12 production by IFN did not seem to have a critical effect during the early stage of IFN therapy. PMID- 14563424 TI - Frequencies of interferon-gamma and interleukin-10 secreting cells in peripheral blood mononuclear cells and liver infiltrating lymphocytes in chronic hepatitis B virus infection. AB - Cytokine balance may play an important role in effective antiviral immunity. We determined the frequencies of interferon-gamma (IFN-gamma)-, interleukin-4 (IL-4) , and interleukin-10 (IL-10)-secreting cells in response to HBV antigens in peripheral blood mononuclear cells (PBMCs) and liver-infiltrating lymphocytes (LILs) using an enzyme-linked immuno spot (ELISpot) assay and related them to serum ALT and HBV DNA levels, and hepatic histological findings. PBMCs were obtained from 25 patients with chronic hepatitis B, from eight of whom LILs were also obtained, and 12 healthy controls. On stimulation with hepatitis B core antigen (HBcAg), the median (range) frequencies of IFN-gamma- and IL-10-secreting cells were 25 (7-71) and 54 (26-101) cells/10(4) PBMCs, respectively, in patients with chronic hepatitis B, and 4 (0-12) and 36 (7-63) cells/10(4) PBMCs, respectively, in healthy controls. The frequencies of HBcAg-specific IFN-gamma secreting cells in PBMCs and LILs of chronic hepatitis B patients correlated with serum ALT levels. Those of LILs correlated with serum ALT levels and HAI scores. In conclusion, HBcAg-specific IFN-gamma-secreting cells may play a role in liver damage in chronic HBV infection. Excessive IL-10 production by PBMCs and LILs in response to HBcAg may suppress antiviral immune responses and contribute to persistent infection. PMID- 14563425 TI - Beneficial effects of branched-chain amino acids on altered protein and amino acid metabolism in liver cirrhosis: evaluation in a model of liver cirrhosis induced in rats with carbon tetrachloride. AB - The effect of branched-chain amino acids (BCAA) on the metabolism of protein, amino acids and ammonia was examined in rats with cirrhosis, with a special emphasis on the efficacy of early administration of BCAA. Liver cirrhosis was induced in rats by intraperitoneal injections of carbon tetrachloride (CCl(4)). The rats were divided into three groups: early-, late- and untreated. The early- and late-treated groups were given BCAA rich food from the early and late stage of liver cirrhosis, while the untreated group was given standard food throughout. Concentrations of amino acids and ammonia in the plasma and the ornithine carbamyl transferase (OCT) activity in the liver were evaluated, and compared with control group after 15 weeks maintenance. Ammonia was significantly higher in the late- and untreated groups, but not in the early-treated group. BCAA, tyrosine, and methionine were significantly lower in the untreated and late treated groups. Glutamine increased significantly in the un-, late- and early treated groups. However, no significant differences were found among three groups. A significant difference was found in 3-methyl histidine (3-MH) between the early- and late-treated groups. This study suggests that the administration of BCAA from early stage of liver cirrhosis inhibits breakdown of protein and improves metabolism of protein, amino acids and ammonia. PMID- 14563426 TI - Can ultrasonographic findings of perihepatic lymphadenopathy promote investigations to detect possible presence of virus C infection in non symptomatic subjects? AB - PURPOSE: Only early detection of non-symptomatic patients is able to arrest the diffusion of the non-symptomatic HCV infection and lead to prompt treatment. Our aim was to attempt to correlate the presence of perihepatic lymph nodes and hepatitis C infection and to assess whether ultrasonography can have a role to promote specific investigations for pre-clinical diagnosis of virus C infection. METHOD: We performed a controlled study on a cohort of 7974 subjects from a town of 27000 inhabitants on the eastern coast of Sicily. Serologic hepatitis A, B and C markers, alanine aminotransferase levels and abdominal ultrasound examination according to size and number of peri-hepatic lymph nodes were performed on blind basis. RESULTS: One or more pathological lymph nodes were present in 684/7974 subjects. Haematochemical tests revealed the presence of anti-HCV positivity in 528/684 subjects with pathological lymph nodes and in 8/7290 subjects without pathological lymph nodes, there being a significant difference (P<0.0001) between the two groups. CONCLUSION: Our results confirm the association between perihepatic lymph nodes and virus C infection. Correct diagnostic assessment of this datum could lead not only to early diagnosis by specific blood test for HCV and consequent prompt aimed treatment, but could pave the way for efficacious territorial prevention and detection of an elevated percentage of likely non symptomatic carriers. PMID- 14563427 TI - Natural course of changes in hepatic functional reserve in patients with chronic liver diseases evaluated by scintigraphy with GSA. AB - Hepatic functional reserve can be evaluated in a noninvasive way by scintigraphy with 99mTc-DTPA-galactosyl human serum albumin (99mTc-GSA). We monitored hepatic functional reserve in patients with chronic hepatitis and cirrhosis using scintigraphy with 99mTc-GSA to determine the natural course of changes in their hepatic functional reserve. Computer acquisition of gamma-camera data was started before the injection of 185 MBq of 99mTc-GSA and was stopped 20 min later. Time activity curves were generated from ROI for the heart and liver. A receptor index and index of blood clearance were calculated from radioactivity in the heart and liver. Scintigraphy with 99mTc-GSA was performed in 12 healthy subjects, 86 patients with chronic hepatitis, and 226 patients with cirrhosis. Seventy-two patients (23 with chronic hepatitis, 32 with cirrhosis in Child-Pugh stage A, 15 in stage B, and 2 in stage C) were examined at least twice with 12-72 months intervening. The receptor index was lower for more severe disorders, decreasing in the order of chronic hepatitis and cirrhosis in stages A, B, and C. The index of blood clearance was higher for more severe disorders, increasing in the order of chronic hepatitis and cirrhosis in stages A, B, and C. The mean annual change in the receptor index with chronic hepatitis was -0.0007, that with cirrhosis in stage A was -0.0023, and that with cirrhosis in stage B or C was -0.0117. The difference between the median annual change with cirrhosis in stage B or C and that with chronic hepatitis or cirrhosis in stage A was not significant (P=0.064 and 0.251, respectively). The mean annual change in the index of blood clearance with chronic hepatitis was 0.0018, that with cirrhosis in stage A was 0.0060, and that with cirrhosis in stage B or C was 0.0330. The difference between the median annual change in the index of blood clearance with cirrhosis in stage B or C and that with chronic hepatitis or cirrhosis in stage A was significant (P=0.004 and 0.007, respectively). Hepatic receptor imaging with 99mTc-GSA could be used to noninvasively evaluate the hepatic reserve of various liver diseases. Changes in hepatic functional reserve were not steady; it decreased gradually as disease advanced from chronic hepatitis to cirrhosis in Child-Pugh stage A, and decrease rapidly after development of stage B cirrhosis. PMID- 14563428 TI - Early development of cavernomatous vasculatures in portal venous thrombosis: morphometric kinetics in rabbit model. AB - We tried to establish an animal model of portal venous thrombosis in order to analyze the ensuing pathological changes of the liver. An emulsion with Escherichia coli endotoxin and Lipiodol-Ultra-Fluide was injected into the portal vein of the left anterior lobe of the rabbit liver. The target lobe and portal vein were time-sequentially examined. In the experimental groups, it was found that fibrin thrombi were formed in the portal vein within 48 h after the injection and thrombi persisted for over 7 days. In an association with thrombus formation, numerous tortuously dilated vasculatures developed in the portal tract (cavernomatous vasculature) within 48 h. Both the number and the total area of the cavernomatous vasculatures increased from 2- to 3-fold more than in the control group at 72 h. The majority of proliferated vasculatures were positive for alpha-smooth muscle actin, thus suggesting that they derived from the portal venous branches. In conclusion, portal endotoxemia may be one of the pathogenetic factors of portal venous thrombosis and, in this model, the cavernomatous vasculatures rapidly developed from the portal venous branches. PMID- 14563429 TI - Activated Kupffer cells play an important role in intra-hepatic Th1-associated necro-inflammation in Concanavalin A-induced hepatic injury in mice. AB - BACKGROUND/AIMS: To examine whether or not activated Kupffer cells play an important role in intra-hepatic Th1-associated necro-inflammation in Concanavalin A (Con A)-induced hepatic injury in mice. METHODS: Con A was administered to Balb/c mice pretreated with or without gadolinium chloride (GdCl(3)). Kupffer cell activation was evaluated by their ability to produce superoxide anions in situ under liver perfusion with nitro blue tetrazolium (NBT). Hepatic concentration of cytokines was measured by ELISA and the mRNA expression of CXC chemokine receptor 3 (CXCR3) was evaluated by RT-PCR. Immunohistochemical detection of CD4 positive lymphocytes in the liver was also performed. RESULTS: GdCl(3)-pretreatment significantly (P<0.01) reduced the serum levels of alanine aminotransferase (ALT) in Con A-treated mice. Formazan deposition in Kupffer cells, the hepatic concentration of tumor necrosis factor-alpha and interferon gamma, the mRNA expression of CXCR3 and the CD4 positive lymphocytes in the liver were decreased in GdCl(3)-pretreated mice as compared with those without GdCl(3) pretreatment (P<0.05, respectively). CONCLUSIONS: Activated Kupffer cells, which produce superoxide anions, are involved in Con A-induced hepatic necro inflammation in mice possibly through the activation of Th1-associated immune response mediated by CD4 and/or CXCR3 positive cells recruited into the liver. PMID- 14563430 TI - Vectorial transport of bile acids in immortalized mouse bile duct cells. AB - In ileal epithelial cells, apical sodium-dependent bile acid transporter (ASBT) is responsible for the uptake of bile acids from the lumen. Furthermore, ASBT is expressed in the apical plasma membrane of intrahepatic bile duct cells (BECs). Using cultured immortalized mouse intrahepatic BECs that form monolayers or cysts, vectorial transport of bile acids was studied. [3H]-taurocholic acid ([3H] TCA) was transported through monolayers transcellularly almost exclusively from the apical to the basolateral side in a Na(+)- and a temperature-dependent manner. Transport of [3H]-TCA was inhibited by 59.3+/-18.6% in the presence of taurochenodeoxycholic acid. Uptake of lysyl fluorescein-conjugated bile acid, Cholyl-[Nepsilon-NBD]-lysine, was seen in a Na(+)- and a temperature-dependent manner from the apical side of BECs that form monolayer or cysts. Reverse transcription-polymerase chain reaction for mRNAs in the cells showed presence of mRNAs for ASBT and farnesoid X receptor (FXR), a nuclear bile acid receptor. In conclusion, intrahepatic BECs transport bile acids mainly from the apical to the basolateral side in concert with ASBT and maybe FXR in the cells. PMID- 14563431 TI - The value of the ultrasonic microprobe in the detection and treatment of rectal varices: a case report. AB - An 81-year-old woman with idiopathic portal hypertension was admitted to our hospital with anal bleeding. Colonoscopy revealed tortuous rectal varices extending 5 cm proximal of the dentate line. The rectal varices were considered to be the most likely cause of the bleeding, although the precise site could not be determined. The vessel images of the intramural rectal varices were clearly delineated using the water repletion method via an ultrasonic microprobe (UMP). The peri-rectal collateral veins and communicating veins between the intramural rectal varices and the peri-rectal collateral veins could be observed via the UMP. We performed endoscopic injection sclerotherapy (EIS) three times using 5% ethanolamine oleate, the total amount of sclerosant used being 5 ml. Ten days after EIS, colonoscopy revealed both ulcers and shrinkage of the varices in the rectum. The UMP revealed the disappearance of variceal vessels and peri-rectal collateral veins. PMID- 14563432 TI - Hepatocellular carcinoma with sarcomatous change arising after radiofrequency ablation for well-differentiated hepatocellular carcinoma. AB - We report a case involving well-differentiated hepatocellular carcinoma (HCC) developing to HCC with sarcomatous changes after radiofrequency ablation (RFA). In a cirrhotic patient with both hepatitis B surface antigen and hepatitis C virus RNA, a well-differentiated HCC with a diameter of 2 cm was detected in segment IV of the liver. A combination of transcatheter arterial embolization and percutaneous ethanol injection (PEI) was performed and, after 8 months, PEI was performed for recurrent tumors. Fifteen months after the first treatment, a recurrent tumor measuring 3.5 cm in diameter was detected in segment IV, which was demonstrated as well-differentiated HCC by tumor biopsy, and treated by RFA. Although the treated lesion was reduced to 2.5 cm in diameter 6 months after RFA, the tumor rapidly enlarged to 6 cm in diameter 2 months later and progressed to lymph node metastasis. Aspiration biopsy showed spindle-shaped sarcomatoid cells with positive staining for both vimentin and keratin. The patient died of HCC progression 10 months after RFA. Autopsy findings showed both sarcomatoid cells and trabecular HCC cells. The sarcomatoid cells had metastasized to the lymph nodes and distant organs. This is the first case illustrating a sarcomatous HCC after RFA. Of interest, RFA may be related to the development of sarcomatous HCC. PMID- 14563433 TI - Optimal clinical implementation of the Siemens virtual wedge. AB - Installation of a modern high-energy Siemens Primus linear accelerator at the Northern Centre for Cancer Treatment (NCCT) provided the opportunity to investigate the optimal clinical implementation of the Siemens virtual wedge filter. Previously published work has concentrated on the production of virtual wedge angles at 15 degrees, 30 degrees, 45 degrees, and 60 degrees as replacements for the Siemens hard wedges of the same nominal angles. However, treatment plan optimization of the dose distribution can be achieved with the Primus, as its control software permits the selection of any virtual wedge angle from 15 degrees to 60 degrees in increments of 1 degrees. The same result can also be produced from a combination of open and 60 degrees wedged fields. Helax TMS models both of these modes of virtual wedge delivery by the wedge angle and the wedge fraction methods respectively. This paper describes results of timing studies in the planning of optimized patient dose distributions by both methods and in the subsequent treatment delivery procedures. Employment of the wedge fraction method results in the delivery of small numbers of monitor units to the beam's central axis; therefore, wedge profile stability and delivered dose with low numbers of monitor units were also investigated. The wedge fraction was proven to be the most efficient method when the time taken for both planning and treatment delivery were taken into consideration, and is now used exclusively for virtual wedge treatment delivery in Newcastle. It has also been shown that there are no unfavorable dosimetric consequences from its practical implementation. PMID- 14563434 TI - Myths and fallacies in permanent prostate brachytherapy. AB - Because there are competing modalities to treat early-stage prostate cancer, the constraints or deficiencies of one modality may be erroneously applied to others. Some valid concerns arising from surgery and external beam therapy, which have been falsely transferred to brachytherapy, are constraints based on patient age, clinical and pathological parameters, patient weight, and size of prostate. Although the constraints have a valid basis in one modality, knowledge of the origin and mechanism of the constraint has provided a means to circumvent or overcome it in brachytherapy. Failures as measured by biochemical no-evidence of disease (bNED) survival may be attributed to extracapsular disease extension. Such extension often expresses itself in surrogate parameters such as a high percentage of positive biopsies, perineural invasion, or the dominant pattern in Gleason score histology. Failures due to such factors may be prevented by implanting with consistent extracapsular dosimetric margins. Some presumed limitations on prostate brachytherapy originated from data on patients implanted in the first few years the procedure was being developed. Most of the urinary morbidity and a significant part of the decrease in sexual function observed may be avoided by controlling the dosimetry along the prostatic and membranous urethra and at the penile bulb. PMID- 14563435 TI - An evaluation of forward and inverse radiotherapy planning using Helax-TMS (version 6.0) for lung cancer patients treated with RTOG 93-11 dose-escalation protocol. AB - This study investigated the dosimetric advantages of inversely planned intensity modulated radiotherapy (IMRT) over forward-planned conventional 3D conformal radiotherapy (3D-CRT) in treating lung cancer patients at escalated dose. Three consecutively accrued patients on the RTOG 93-11 dose-escalation protocol were replanned using IMRT with the same dosimetric rules, so that the isodose distributions and dose-volume histograms could be generated and compared. The Helax-TMS treatment planning system, with an IMRT optimization module (version 6.0), was used. In all cases, a consistent approach of inverse planning and set of dose-volume constraints (DVCs) provided improved critical structure sparing. However, the minimum dose in PTV was generally below that achieved with the corresponding forward planned 3D-CRT. PMID- 14563436 TI - A 3D technique for simulation of irregular electron treatment fields using a digital camera. AB - Cerrobend inserts, which define electron field apertures, are manufactured at our institution using perspex templates. Contours are reproduced manually on these templates at the simulator from the field outlines drawn on the skin or mask of a patient. A previously reported technique for simulation of electron treatment fields uses a digital camera to eliminate the need for such templates. However, avoidance of the image distortions introduced by non-flat surfaces on which the electron field outlines were drawn could only be achieved by limiting the application of this technique to surfaces which were flat or near flat. We present a technique that employs a digital camera and allows simulation of electron treatment fields contoured on an anatomical surface of an arbitrary three-dimensional (3D) shape, such as that of the neck, extremities, face, or breast. The procedure is fast, accurate, and easy to perform. PMID- 14563437 TI - Patient setup and verification for intensity-modulated radiation therapy (IMRT). AB - Intensity Modulated Radiation Therapy (IMRT) is now widely used in the radiation therapy community. The ability of IMRT to deliver complex dose distributions has allowed dose escalation to targets while sparing normal tissues. In IMRT the roles of the physicist, dosimetrist, and physician are changed. Inverse planning, which is inherent to IMRT, requires that the final dose solution be defined at the beginning of the planning process. The physician must define specific dose volume constraints for the target as well as normal tissues. The physicist and dosimetrist must evaluate the final plan and determine if it meets the goals of the treatment, even if it does not completely satisfy the initial constraints. Once a plan is decided upon, the ability of the clinic to safely and accurately deliver that plan to the patient must be confirmed. As with any new technology, IMRT has created a need for new quality assurance procedures. Here we describe our IMRT process from simulation through planning and treatment. By standardizing our simulations we have decreased setup times and decreased the threat of collisions. Comparison of pseudo-DRR's and multiple-exposure port films allows confirmation of patient positioning on the linac. Our treatment delivery quality assurance program using film and MOSFET detectors in a polystyrene phantom is also described. We provide insight on how to overcome some of the common problems encountered in treatment planning and delivery such as isocenter location, collision avoidance, table indexing, dose confirmation, and plan analysis. PMID- 14563439 TI - Various techniques of contouring the rectum and their impact on rectal dose volume histograms. AB - Late rectal bleeding resulting from radiotherapy has been correlated with rectal dose-volume histograms (DVHs). The techniques of contouring the rectum have been inconsistent within the literature, making interpretations of DVHs difficult. This study was conducted to investigate the impact on rectal DVHs when using different ways of contouring. Ten prostate cancer patients were treated with a 4 field box-technique and received 70 Gy. Six different ways of contouring the rectum were implemented by using 3 different cross-sections and 2 different lengths. The 3 different cross sections were (1) anterior rectal wall (arw): only the anterior half of the rectal wall was contoured; (2) whole rectal wall (wrw): the entire rectal wall was contoured but excluding the rectal contents; and (3) the rectum (rec): including the rectal contents. Two different lengths were used for the above 3 volumes: (1) long (Lg): cranial border starting at where the rectum turned horizontally into the sigmoid and the caudal border 2 cm below the prostatic apex; (2) short (Sh): from 2 cm above to 2 cm below the prostate. Therefore, a total of 6 different volumes (Sh arw, Lg arw, Sh wrw, Lg wrw, Sh rec, and Lg rec) were generated. DVHs of all 6 volumes were compared with the y axis being percentage volume as well as absolute volume (cc). When using percentage volume as the y-axis, Sh arw gave an impression that a large portion of rectum (median of 41.8%) received high dose (greater than 90% prescribed dose), while the Lg wrw and Lg rec revealed a smaller portion of rectum (median of 17.1% and 14.7%, respectively) received high dose. The other contours were somewhere in-between. When using absolute volume as the y-axis, the DVHs of the 4 rectal volumes, excluding the rectal content (Sh and Lg arw, Sh and Lg wrw), merged at doses greater than 80% to 85% prescription, therefore providing similar information within these high-dose regions. Configurations of rectal DVHs varied drastically with different techniques of contouring and may lead to different interpretations. By using absolute volume (cc) as the y-axis, the shape of the 4 rectal DVHs, excluding the rectal content, were similar in the high-dose region. Reporting rectal toxicities in relations to DVHs using absolute volume, as well as percentage volume, may eliminate inconsistencies secondary to different methods of contouring. PMID- 14563438 TI - Prostate implant nomograms for the North American Scientific 103Pd seed. AB - Palladium-103-(103Pd) seed has been increasingly used in prostate implantation as either definitive or boost therapy because of its shorter half-life and higher initial dose rate. Because a growing number of radiation oncologists prefer real time implantation in the operating room, it will be helpful if the total activity of the seeds can be determined based on the gland size before the patient is taken to the operating room. Based on our clinic data, nomograms have therefore been developed for one of the widely used 103Pd seeds, the MED3633 seed, which is produced by North American Scientific, Inc. (NASI). The total activities for implant volume ranging from 15 cc to 55 cc are provided for both seed "monotherapy" and seed boost. PMID- 14563440 TI - Determination of surface dose and the effect of bolus to surface dose in electron beams. AB - When treating tumors from surface to a certain depth (<5 cm), electron beams are preferred in radiotherapy. To increase the surface doses of lower electron beams, tissue-equivalent bolus materials are often used. We observed that the surface doses increased with increasing field sizes and electron energies. At the same time, we also observed that all electron parameters were shifted toward the skin as much as the thickness of the bolus used. The effect of bolus to the surface doses was more significant at low electron energies than at higher electron energies. Rando phantom measurements at 6-, 7.5-, and 9-MeV were slightly lower than the solid phantom measurements, which could only be explained by the inverse square law effect and the Rando phantom contour irregularity. PMID- 14563441 TI - Triglycerides and small dense LDL: the twin Achilles heels of the Friedewald formula. PMID- 14563442 TI - Natural abundance 13C-NMR spectroscopy for the quantitative determination of fecal fat. AB - OBJECTIVE: To evaluate 13C-NMR spectroscopy as a method for fat quantitation in human feces without time consuming or unpleasant preparation steps. DESIGN AND METHODS: Stool samples of seven healthy subjects were collected for 18 days before and during oral intake of the inhibitor of gastrointestinal lipases Orlistat. Fecal lipid content was determined first using 13C-NMR, then by conventional gravimetry after homogenization and Bligh & Dyer lipid extraction. RESULTS: The correlation between gravimetry and 13C-NMR was excellent (R2 = 0.91). In repeated measurements, the mean percentage error was 2.8%. On average, 13C-NMR yielded 1.27 g less fat than gravimetry. Orlistat efficacy for fat excretion assessed by 13C-NMR and by gravimetry was 34.3% and 33.9%, respectively. CONCLUSIONS: With a total measurement time of three minutes, 13C NMR spectroscopy of unprocessed whole stool provides an accurate alternative to gravimetry for assessing total fecal fat excretion. 13C-NMR is superior with regard to practicability and speed. PMID- 14563444 TI - Influence of anticoagulants on the measurement of S100B protein in blood. AB - OBJECTIVE: Evaluate anticoagulants influence on plasma S100B levels. DESIGN AND METHODS: Blood were collected from 18 healthy adult subjects using: no anticoagulants, EDTA, heparin, and citrate. S100B levels were determined using LIA-mat assay. RESULTS: Heparin plasma and citrate increased plasma S100B levels (p < 0.001), whereas EDTA had no effect (p = 0.24). Heparin plasma samples were highly (r2 = 0.97, p < 0.001), citrate samples were moderately (r2 = 0.49, p < 0.001), and EDTA samples were not (r2 = 0.22, p = 0.059) correlated with serum samples. CONCLUSIONS: When anticoagulant is required, heparin plasma should be the primary choice for measurement of S100 B levels. PMID- 14563443 TI - Detection of the four sequence variations of MDR1 gene using TaqMan MGB probe based real-time PCR and haplotype analysis in healthy Japanese subjects. AB - OBJECTIVES: P-glycoprotein (P-gp) is significant from the viewpoint of pharmacokinetics/pharmacodynamics (PK/PD). MDR1 gene encodes P-gp and has a wide variety of SNPs. As the SNPs may be one of the factors that induce pharmacogenetic individual difference, haplotype analysis is necessary to evaluate the PK/PD. DESIGN AND METHODS: The SNPs of the detected MDR1 were 129T>C, 325G>A, 2677G>T/A, and 3435C>T. For the analysis of linkage disequilibrium (LD) and haplotype analysis, and for the reconstruction of the haplotype pair, ARLEQUIN and PHASE were employed. RESULTS: The result of the chi(2) test detected significant LD between -129 and 2677, -129 and 3435, and 2677 and 3435. There were 9 haplotypes: T-G-C, T-T-C, C-T-C, T-A-C, C-A-C, T-G-T, T-T-T, C-G-T, and C-T-T. CONCLUSIONS: LD was found among the positions -129, 2677 and 3435. As a result, 9 haplotypes exists in the Japanese population. These results suggest that it would be necessary to give consideration to haplotype for the purpose of evaluating the PK/PD of the drugs transported by P-gp. PMID- 14563445 TI - Two-center clinical evaluation of a new automated fluorometric immunoassay for the quantitative analysis of total betaeta-human chorionic gonadotropin. AB - OBJECTIVES: This study evaluated the quantitative total betahCG assay on the Stratus CS point-of-care instrument at two medical centers. DESIGN AND METHODS: Analytical sensitivity, linearity, within-run and total imprecision, interferences, dilution recovery, method comparison (Dimension RxL), comparison of matched heparinized whole blood and plasma samples, and determination of the normal reference interval were studied. RESULTS: Analytical sensitivity was <0.5 IU/L. The assay's linear range was 0 to 1250 IU/L; the clinical reportable range was up to 50,000 IU/L. Within-run imprecision (CV) at both low (<20 IU/L) and elevated (760 IU/L) betahCG concentrations were <4%. Total imprecision for three QC levels and two pools were <4%. Method comparison showed Stratus CS betahCG = 0.98 +/- 0.01* Dimension RxL hCG -0.11 +/- 2.69 (n = 136; r = 0.996; Sy/x = 27.7). Matched heparinized whole blood/plasma sample-comparison showed: whole blood = 1.05*Plasma + 0.37 +/- 1.29 (n = 41; r = 1.000; Sy/x = 7.57). Mean dilution recovery was 99% (range: 95% to 103%). None of the 52 drugs tested, lipemia, icterus, hemolysis, LH, FSH, TSH, hGH or prolactin represented a significant interference with the assay. Reference intervals were <0.5 IU/L for males (n = 123) and <3.0 IU/L for nonpregnant females (n = 120). CONCLUSIONS: The Stratus CS betahCG test offers the advantage of quantitative measurement of total betahCG in whole blood at the point of care and is suitable for clinical use. PMID- 14563446 TI - Intestinal-type and liver-type fatty acid-binding protein in the intestine. Tissue distribution and clinical utility. AB - OBJECTIVES: Intestinal-type fatty acid-binding protein (I-FABP) has been proposed as plasma marker for the detection of acute intestinal injury. However, intestinal mucosa also expresses liver-type FABP (L-FABP). We have investigated the tissue distribution of I-FABP and L-FABP in segments of the human intestine along the duodenal to colonal axis and the potential of both proteins to serve as plasma marker for the diagnosis of intestinal injury. DESIGN AND METHODS: I-FABP and L-FABP were measured with specific immunoassays in autopsy samples of the intestine (duodenum, jejunum, ileum and colon) of 23 subjects and in plasma samples from patients (n = 51) with intestinal and/or hepatic disease. Plasma reference values were established in normal healthy individuals (n = 92). RESULTS: The I-FABP tissue contents in duodenum, jejunum, ileum, proximal colon and distal colon amounted to 2.22, 4.79, 1.04, 0.27 and 0.25 mug/g ww, respectively. L-FABP tissue contents were markedly higher, amounting to 124 and 198 mug/g ww in duodenum and jejunum, and to 58, 26 and 44 mug/g ww in ileum, proximal colon and distal colon, respectively. Elevated plasma levels of both I FABP and L-FABP were found in patients suffering from intestinal diseases, while only L-FABP was increased in cases of purely hepatocellular injury. CONCLUSIONS: I-FABP and L-FABP show a similar pattern of tissue distribution along the duodenal to colonal axis with highest tissue contents found in the jejunum but in each intestinal segment a >40-fold higher content of L-FABP than of I-FABP. Accordingly, besides I-FABP, also L-FABP is a useful plasma marker for the detection of intestinal injury, especially in patients undergoing intestinal surgery. PMID- 14563447 TI - Detection of urokinase plasminogen activator receptor and c-erbB-2 in sera of patients with breast and ovarian carcinoma. AB - The role of urokinase plasminogen activator receptor (uPAR) and c-erbB-2 in breast and ovarian cancer was investigated. Eighty patients of breast and ovarian cancer and benign lesions, as well as twenty normal controls were evaluated for the expression of c-erbB-2 by Western blotting and uPAR levels by ELISA. The c erbB-2 and uPAR showed a significant increase in both types of cancer investigated compared to normal control and benign lesions. The frequency of c erbB-2 was significantly higher in breast cancer lesions (p < 0.01). Levels of CA15.3 in breast cancer and CA125 in ovarian cancer were significantly higher in cases expressing c-erbB-2 (p < 0.01) than in negative c-erbB-2 cases. The uPAR showed a significant positive correlation with advanced stages of breast cancer (r = 0.7971) and ovarian cancer (r = 0.83662), while significant correlations were found for CA15.3 in breast cancer (r = 0.64967) and CA125 in ovarian cancer (r = 0.83996). Taken together, our data suggest that the c-erbB-2 and uPAR in the sera of ovarian and breast cancer act as valuable markers for the evaluation of the patients preoperatively. PMID- 14563448 TI - Apoprotein(a) phenotypes and plasma lipoprotein(a) concentration in patients with diabetes mellitus. AB - OBJECTIVES: To determine whether apo(a) isoforms and plasma Lp(a) concentrations in association with some lipid parameters increase the relative risk for the development of atherosclerosis in patients with diabetes mellitus (IDDM and NIDDM). DESIGN AND METHODS: Apo (a) isoforms, Lp(a) and plasma lipids were determined in 40 IDDM and 65 NIDDM patients and in 182 healthy individuals. Apo(a) isoforms were separated by 3 to 15% gradient SDS-PAGE followed by immunoblotting. RESULTS: Logistical analysis showed that: Lp(a) levels >30 mg/dL (RR = 0.25, p < 0.000001; RR = 0.18, p < 0.00002), HTA (RR = 0.212, p < 0.00001; RR = 0.30, p < 0.00001), LMW-S1 apo(a) isoform (RR = 6.86, p < 0.0131; RR = 7.04, p < 0.0057) play a significant role in aterogenecity in both groups of patients with DM (IDDM and NIDDM). The 6.50-fold increase in risk was found in NIDDM patients with high Lp(a) levels (>30 mg/dL) and plasma total/HDL cholesterol ratio (4.5-5.8). CONCLUSION: Elevated Lp(a) levels, LMW S1 apo(a) isoform, HTA and combination of increased Lp(a) levels and total/HDL cholesterol ratio increase the risk for the development of atherosclerosis in patients with DM (IDDM and NIDDM). PMID- 14563449 TI - Serum markers of monocyte/macrophage activation in patients with Alzheimer's disease and other types of dementia. AB - OBJECTIVES: Recently an increase in serum neopterin has been described in patients with Alzheimer's disease (AD) that would be associated with an increased cell-mediated immune response. We have studied the serum levels of several monocyte/macrophage activation markers in patients with AD and other types of dementia. DESIGN AND METHODS: Serum neopterin concentration, and the chitotriosidase (ChT), angiotensin-converting enzyme (ACE) and adenosine deaminase (ADA) activities were determined in 30 patients with AD, in 19 patients with other types of dementia, and in 24 nonaffected controls. RESULTS: Neopterin concentration was significantly higher in the subgroup of AD patients with a global deterioration scale higher than in the other patients with AD, patients with other types of dementia and in the control group (p < 0.005). However, the activities of ChT, ACE and ADA, despite having a significant correlation with neopterin, did not present any statistically significant differences among the groups studied. CONCLUSION: In the most advanced clinical stages of AD, as well as an increased immune activation, an impaired formation of tetrahydrobiopterin from dehydroneopterin triphosphate would contribute to an increase in the serum concentration of neopterin. However, the large overlap between the groups, limits the possible clinical value of serum neopterin in AD patients. PMID- 14563450 TI - Antioxidant status, lipid peroxidation and nitric oxide end products in patients of type 2 diabetes mellitus with nephropathy. AB - OBJECTIVES: Oxidative stress is considered to be a unifying link between diabetes mellitus (DM) and its complications including nephropathy. The aim of the present study was to evaluate oxidative stress status in Asian Indian patients of type 2 DM with nephropathy. DESIGN AND METHODS: Serum levels of malondialdehyde (MDA) and nitric oxide end products (nitrite and nitrate), activities of erythrocyte superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) content were estimated in controls, patients of type 2 DM without nephropathy (group 1) and with nephropathy (group 2). RESULTS: Serum MDA concentration was significantly high in both the groups of diabetic patients as compared to controls, (p < 0.05), with group 2 having a significantly higher value than group 1 (p < 0.05). Significantly elevated serum nitrite levels were found in diabetic patients as compared to controls (p < 0.001), however, no significant difference was found between group 1 and group 2. Moreover, serum nitrate as well as nitrite + nitrate levels were significantly higher in group 2 as compared to controls (p < 0.05). Activity of erythrocyte SOD and CAT was significantly reduced in both groups as compared to controls (p < 0.001) with catalase activity in group 2 being significantly lower than group 1 (p < 0.05). Erythrocyte GSH content was significantly lower in group 2 as compared to controls (p < 0.05) and group 1 (p < 0.05). CONCLUSIONS: Results of the present study indicate that oxidative stress is increased and antioxidant defenses are compromised in type 2 DM. These derangements are of a higher magnitude in patients of type 2 DM with nephropathy. PMID- 14563451 TI - Lipoprotein (a) and lipid profile in neonates from mothers with three different types of diabetes mellitus. AB - OBJECTIVES: The metabolic components in neonates may be affected by maternal diabetes mellitus. DESIGN AND METHODS: To investigate the alterations in lipid metabolism and the possible atherogenic risk, the lipoprotein a (Lp a), apoproteins, lipid profile, glucose concentrations were measured (ELISA, immunodiffusion and enzymatic) in 77 cord blood samples from diabetic and healthy pregnant mothers. RESULTS: The body weight, cord glucose and both apoproteins were increased in neonates of gestational and noninsulin dependent diabetic (GDM, NIDDM) than in neonates of nondiabetic mothers (NNDM). The Lp (a) was not correlated with the blood glucose and didn't significantly increase in the three neonates groups of diabetic mothers. The apo B/apo A1 and the LDL/HDL ratios were insignificantly increased in relation to the body weight. In neonates of diabetic mothers (NDM), only the blood glucose and Lp (a) differ between both sexes. CONCLUSION: NDM may have disturbed lipid metabolism, which require special care to them and to their mothers during the prenatal period. PMID- 14563452 TI - Hyperlipidemic profiles during remission in childhood idiopathic nephrotic syndrome. AB - Hyperlipidemia, an important characteristic of idiopathic nephrotic syndrome in children (NS), is usually observed during the active phase of the disease and disappears with the resolution of the proteinuria. However, persisting lipid anomalies during remission have been reported in a few studies and raise the question of the later development of atherosclerosis. Plasma lipid profiles in 25 children with NS at remission, with or without active prednisone treatment, were compared with those of an age-matched population. The results indicate that plasma total and LDL-cholesterol levels were above the 95(th) percentile for age and sex in 12 of the 25 patients (48%) with 7 of them having apolipoprotein B and triglyceride concentrations above the 95(th) percentile. Moreover, frequently relapsing children were more likely to have abnormal lipid profile during the remission. We conclude that close monitoring of lipid levels during the remission of the NS especially in those with frequent relapses, is necessary to select the high-risk patients. PMID- 14563453 TI - Plasma lipid and lipoprotein concentrations in pregnancy induced hypertension. AB - OBJECTIVES: Aim of this study was to evaluate implication of pregnancy induced hypertension on maternal plasma lipid, lipoprotein, apolipoprotein concentrations and lipid peroxidation products by a comparison of normal pregnancy vs. preeclampsia. DESIGN AND METHODS: Thirty-four women with preeclampsia and 32 healthy pregnant women (controls) in the third trimester were recruited for this study. RESULTS: In the preeclamptic group plasma total triglyceride, low density lipoprotein cholesterol (LDL-C), malondialdehyde (MDA) and apolipoprotein B (apo B) were significantly increased, while plasma high density lipoprotein cholesterol (HDL-C) was significantly decreased compared to that of control group. There was no significant difference in total cholesterol and apolipoprotein A1 (apo-A1) concentrations. CONCLUSION: Our findings suggest that preeclampsia share some metabolic characteristics with coronary artery disease such as dislipidemia and increased lipid peroxidation. However lipoprotein concentrations may be better biochemical markers of dislipidemia in the preeclamptic state than the corresponding apolipoproteins. PMID- 14563454 TI - A preliminary evaluation of the interaction between urine specific gravity and leukocyte esterase results using Bayer Multistix and the Clinitek 500. AB - OBJECTIVES: To determine the sensitivity of the leukocyte esterase (LE) pad to predict microscopically quantified white blood cells in urine sediment and to assess the interaction of urine specific gravity (SG) on leukocyte esterase sensitivity. METHODS: Microscopic quantification of white blood cells in urine sediment was performed manually and automated chemical urinalysis was performed using the Bayer Clinitek 500 and Multistix 8 SG strips on 422 random urine specimens. RESULTS: The sensitivity of the leukocyte esterase pad to predict white blood cells in centrifuged urine was 58% (61/106). There was a weak but statistically significant negative correlation between SG and LE pad performance, r = -0.12, p < 0.05. CONCLUSIONS: The 58% sensitivity of the Multistix leukocyte esterase detection of white blood cells in urine sediment was consistent with previous reports. The negative correlation of specific gravity and leukocyte esterase provides partial explanation for the lack of sensitivity of the leukocyte esterase test. PMID- 14563456 TI - Parathyroid hormone therapy for osteoporosis. PMID- 14563457 TI - Pondering the precautionary principle. PMID- 14563458 TI - Recent data on the role for angiogenesis in rheumatoid arthritis. AB - Angiogenesis is central to the development and perpetuation of rheumatoid synovitis. Vascular endothelial growth factor (VEGF), the main mediator of angiogenesis, is found in the synovial fluid and serum of patients with rheumatoid arthritis (RA), and its expression is correlated with disease severity. Compelling evidence that VEGF is involved in synovitis has been obtained from experimental models of RA. In particular, VEGF inhibition by synthetic compounds (e.g. TNP-470) or by naturally occurring factors (e.g., the soluble VEGF receptor) produce therapeutic effects. Angiopoietin-1, a recently discovered growth factor specific for neovascularization, is expressed within the rheumatoid synovium and may be stimulated by TNF-alpha. Other compounds, including integrins, fibroblast growth factor, and proinflammatory cytokines contribute to joint angiogenesis and, therefore, to the development of rheumatoid synovitis. Assessing vascularity may prove useful for evaluating or even predicting bone destruction. Furthermore, inhibition of angiogenesis may prove useful as an adjunct to current anti-inflammatory treatments. PMID- 14563459 TI - Musculoskeletal manifestations in cystic fibrosis. AB - Although bone and joint manifestations are common in children with cystic fibrosis (CF), they have received little attention in adults. As compared to healthy individuals, bone mineral density is low, even with calcium intakes greater than 1500 mg/d. Nevertheless, calcium and phosphate levels in blood and urine are often normal, and vitamin D levels vary. Short stature with a low body mass index and central hypogonadism are the rule in these patients. Fractures and kyphosis are often reported. CF arthropathy occurs in 2-8.5% of patients. Arthritis develops, and there may be skin eruptions. Non-steroidal antiinflammatory drug therapy is effective. Hypertrophic osteoarthropathy associated with respiratory failure is present in 2-7% of patients. Rheumatoid arthritis, spondyloarthropathies, sarcoidosis, and amyloidosis have been reported in association with CF. Knee pain due to patellofemoral syndrome, quinolone induced arthropathy, and mechanical back pain have been described. Rheumatoid factor titers are higher than in healthy controls, particularly in patients with episodic arthritis. No data are available on antiperinuclear factor or antikeratin antibody titers. Tests for antinuclear antibody are usually negative. Circulating immune complex levels and antibodies to heat shock proteins may be elevated. Antineutrophil cytoplasmic antibody of the bactericidal/permeability increasing protein (BPI) or azurocidin (AZ) type has been reported, often in high titers (up to 40%). PMID- 14563460 TI - Mechanism of action of spinal manipulative therapy. AB - Spinal manipulative therapy (SMT) acts on the various components of the vertebral motion segment. SMT distracts the facet joints, with faster separation when a cracking sound is heard. Intradiscal pressure may decrease briefly. Forceful stretching of the paraspinal muscles occurs, which induces relaxation via mechanisms that remain to be fully elucidated. Finally, SMT probably has an inherent analgesic effect independent from effects on the spinal lesion. These changes induced by SMT are beneficial in the treatment of spinal pain but short lived. To explain a long-term therapeutic effect, one must postulate a reflex mechanism, for instance the disruption of a pain-spasm-pain cycle or improvement of a specific manipulation-sensitive lesion, whose existence has not been established to date. PMID- 14563461 TI - Tears in the subscapularis tendon: descriptive analysis and results of surgical repair. AB - Although not well known, lesions of the subscapularis tendon are more common than usually reported. We conducted a descriptive study of 22 patients (23 shoulders), who underwent surgical repair of subscapularis tendon tears. Mean follow-up was 73 months. The findings were compared to the results of a prospective study published by one of us (R.H.C.), who reviewed 97 other patients (105 shoulders) with a mean follow-up of 13.4 years after surgical repair of a rotator cuff tear. This comparison allowed us to identify the specific features of rotator cuff tears involving the subscapularis tendon. Subscapularis tears were more likely to be related to a shoulder injury and caused more severe functional impairment, as compared to lesions of the other tendons. Time to surgical treatment was shorter when the subscapularis was torn. Subscapularis tears required adaptation of the surgical approach and postoperative protection of external rotation for 4-6 weeks. Outcomes were less satisfactory when the subscapularis tendon was torn. However, the outcomes in both studies were correlated to tear size. The outcomes were often favorable after isolated subscapularis tendon tears and less so when other tendons were involved. PMID- 14563462 TI - Inverse correlation of each functional status scale of the SF-36 with degree of disease activity in systemic lupus erythematosus (m-SLAM). AB - OBJECTIVES: To compare systemic lupus erythematosus (SLE) disease activity measured by a modified Systemic Lupus Activity Measure (m-SLAM) with functional/health status measured by the SF-36 questionnaire. PATIENTS AND METHODS: m-SLAM and SF-36 scores were obtained on 71 SLE patients during 242 clinic visits over 15 months. Patients were stratified into disease activity groups (m-SLAM <2 = remission; 2-4 = mild; 4-6 = moderate; >6 = severe). Mean SF 36 group scores were compared by analysis of variance (ANOVA). RESULTS: Two hundred and nineteen m-SLAM and SF-36 scores were completed. The disease activity groups correlated inversely with the SF-36 scores in all eight subscales, i.e. the patients' perceived health, as assessed by the SF-36, correlated with their disease activity level as measured by the m-SLAM. Inverse correlation of SLAM activity groups with all eight SF-36 subscales was highly statistically significant. CONCLUSION: The significant inverse correlation of the m-SLAM with all domains of the SF-36 in this study provides potentially useful information for evaluating patients with SLE. PMID- 14563463 TI - Hydatid disease of bone. Review of 11 cases. AB - Osseous hydatid disease is defined as development within bone of multiple cysts that are the larval form of the tapeworm Echinococcus granulosus. Bone cysts account for only 0.5-2.5% of all hydatid cysts in humans. We report on our experience. METHODS: This retrospective study included patients managed between 1988 and 1998 for histologically documented hydatid disease of bone. RESULTS: Eleven patients were included, six men and five women, with a mean age of 40.7 +/ 11.04 years (range, 27-60 years). Mean time to diagnosis was 22.7 +/- 18.2 months (range, 5-36). The pelvis was involved in six patients and a long bone in five (the femur in three, the tibia in one, and the fibula in one). Peripheral eosinophil counts were high in four patients and serological tests for hydatid disease were positive in five patients. Surgical treatment was used alone in 10 patients and with albendazole in one patient. The main complications were fistulization (n = 6) and suppuration (n = 4). Five patients experienced recurrences. CONCLUSION: Hydatid disease still occurs in Morocco. The liver and lungs are the most common targets. Bone cysts are uncommon but severe. The behavior of osseous hydatid cysts resembles that of locally malignant lesions. Although immunofluorescent assays are useful, the final diagnosis depends on histology. The treatment rests on surgical excision. Recurrence is common, particularly at sites that are difficult to access. PMID- 14563465 TI - Pain management in a rheumatology department: a satisfaction survey. AB - OBJECTIVES: To evaluate the quality of pain management in a teaching hospital rheumatology department. METHODS: We conducted a satisfaction survey among all the patients admitted to the conventional rheumatology department of our teaching hospital over a 1-month period (88 patients with a mean length of stay of 5 d). The patients were asked to complete a questionnaire on the day of discharge. The professional staff was not informed of the survey. RESULTS: The mean pain severity score at admission (visual analog scale, VAS) was 7.76 +/- 1.76 and the mean score decrease with treatment was 7.27 +/- 2.81. Expected pain relief and actual pain relief were correlated (R = 0.39; P = 0.001). Nearly all the patients (96.1%) reported have been encouraged to communicate about their pain. Information on the treatment was given to 89.3% of the patients; no significant differences in pain severity or pain relief were found between the patients who did and did not receive this information. The patients were satisfied with their management by the physicians (VAS: 8.83 +/- 2.07) and nurses (VAS: 8.68 +/- 1.72). CONCLUSION: Satisfaction with pain management (a subjective criterion) was good in our patients. However, no validated tools for measuring satisfaction are available, and measurements should be repeated to look for improvements over time. Limitations to these results include the placebo effect, the influence of memory, and the effects of the behavior of hospitalized patients. A repeat survey is needed. PMID- 14563464 TI - Multicenter, randomized, double-blinded, placebo-controlled trial of thiocolchicoside in acute low back pain. AB - OBJECTIVE: To evaluate the efficacy of intramuscular injection of thiocolchicoside (4 mg-2 ml) compared to placebo administered twice daily for 5 days in patients suffering from acute low back pain. PATIENTS AND METHODS: Multicenter, randomized, double-blinded, placebo-controlled trial. The study was conducted between July 1998 and March 2000 in five centers. Hospitalized patients with acute low back pain were included. The primary evaluation criterion was spontaneous pain at rest assessed by visual analog scale (VAS). Hand-to-floor distance, muscle spasm intensity, patients' global evaluation and analgesic tablet (paracetamol) consumption were used as secondary evaluation criteria. RESULTS: One hundred and forty nine patients were included. Both groups showed improvement on spontaneous pain assessed by VAS at the end of day 1; however, improvement was statistically significant in thiocolchicoside group on day 3 (P < 0.001). Hand-to-floor distance and muscle spasm determined by palpation decreased significantly on day 5 in thiocolchicoside group (P < 0.0005 for both). According to the patients' global evaluation, 76.8% of patients in thiocolchicoside group evaluated the treatment as very good/good (P < 0.0005). Also a significant difference was observed in favor of the thiocolchicoside group in the total paracetamol consumption (P < 0.001). Treatment was well tolerated in both groups and none of the patients was withdrawn from the study due to an adverse effect. CONCLUSION: Twice daily administration of 4 mg thiocolchicoside for 5 days provides an efficient and safe treatment for patients with acute low back pain accompanied by muscle spasm. PMID- 14563466 TI - Effects of mud pack treatment on skin microcirculation. AB - OBJECTIVES: The objective of this study was to conduct a laser-Doppler flowmetry investigation of skin microcirculation changes induced by mud pack therapy. The magnitude of the changes, potential remote effects, and potential influence of mud pack thickness were studied. METHODS: Twenty female spa therapy patients aged 28-67 years (median, 51 years) participated in the study. The reason for spa therapy was lower limb venous insufficiency in 14 patients and osteoarthritis in six patients, none of whom had involvement of the shoulders. Mud pack treatment was associated with a significant elevation in skin temperature, by 1.8 +/- 0.2 and 1.0 +/- 0.2 degrees C with the 30- and 15-mm packs, respectively (P < 0.001 for both comparisons; nonsignificant difference between the two packs). Skin blood flow increased significantly, by 619 +/- 82 and 410 +/- 124 mV with the 30- and 15-mm packs, respectively (P < 0.0001 for both comparisons; nonsignificant difference between the two packs). The vasomotion score increased markedly on the treated side, by 16.7 +/- 2.8 and 13.0 +/- 1.6 with the 30- and 15-mm packs, respectively (P < 0.005 for both comparisons; no significant difference between the two packs). Furthermore, low-frequency vasomotion waves of a type not described previously were recorded. The microcirculatory changes lasted longer than did the temperature increase. No significant changes were noted in the other shoulder or in central body temperature. RESULTS: The patients were volunteers receiving spa therapy and free of diabetes mellitus, vasoactive drug treatment, and inflammatory shoulder disease. Two mud packs, 15 and 30 mm in thickness, respectively, were applied at an interval of 48 h, at the same time of day in a given patient, and at a distance from other spa treatments. The packs were centered on the deltopectoral groove. The side and order of application of the two packs were determined at random. Superficial skin blood flow was measured by laser-Doppler flowmetry (Perimed PF4001, wavelength 82 nm) and recorded by the Perisoft computer program with a 3-s time constant downstream from a broadband filter (12 MHz). CONCLUSION: These results suggest that the vascular changes induced by mud pack therapy are not fully explained by vasodilation in response to local temperature elevation. Further studies are in order to identify the other mechanisms involved. PMID- 14563467 TI - Membranous lipodystrophy. Case report and review of the literature. AB - Membranous lipodystrophy (ML) is a rare hereditary disorder of adipose tissue characterized by polycystic bone lesions and progressive dementia. We describe the case of a 36-year-old woman with mechanical bone pain. Routine laboratory analyses revealed only a type IV hyperlipoproteinemia and hyperexcretion of urinary calcium. Roentgenograms of short and long bones showed symmetrical, well defined, non-expansile cystic lesions. Bone biopsy found a yellow lipid-like substance in the osteolytic lesions and histopathological studies were non specific. Neuropsychiatric examination, including cranial computerized tomography (CT), was found to be normal. According to clinical, analytical, radiological and histological findings ML was the diagnosis. No previous cases of ML have been reported in our country as we review the literature concerning this disease. PMID- 14563468 TI - JC virus leukoencephalopathy complicating Wegener's granulomatosis. AB - Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease of the brain caused by the JC virus. It occurs in association with immunodepression due, for instance, to a hematological malignancy, HIV infection, or immunosuppressive therapy for an organ transplant or systemic disease. We describe the fourth reported case of PML in a patient receiving immunosuppressants for Wegener's granulomatosis. A 71-year-old woman receiving azathioprine and glucocorticoid therapy experienced onset of right-sided hemiplegia within a few days, became comatose, and died within a few days. MRI of the brain showed a subcortical lesion in the left parietal lobe generating low signal on T1 images and high signal on T2 images. The initial diagnosis was cerebral vasculitis. However, the postmortem examination showed PML. The diagnosis of PML rests on JC virus detection in the cerebrospinal fluid by PCR assay and on demonstration in a brain biopsy of the typical histological pattern with presence of the JC virus within the demyelinated lesions. No specific or effective treatments are available. Immunosuppressant drugs should be discontinued if possible. PMID- 14563469 TI - A case of ANCA-associated vasculitis with predominant involvement of central nervous system. AB - The authors present a peculiar case of a 53-year-old woman affected by ANCA associated vasculitis with prevailing involvement of the central nervous system. Diagnosis was really difficult. PMID- 14563470 TI - Vascular manifestations of Behcet's disease. Eighteen cases among 140 patients. AB - OBJECTIVES: To describe the features, prognosis, and treatment of vascular involvement in Behcet's disease (BD). PATIENTS: Among 140 patients with BD seen at the Hotel-Dieu Hospital in Beirut between 1980 and 2000, 18 (13%) had vascular involvement and were included in this retrospective study. All these patients fulfilled International Study Group criteria for BD. RESULTS: Men with BD were more likely to have vascular involvement (13/77, 17%) than women (5/63, 8%) (P = 0.12) and were younger at diagnosis of vascular disease (32 +/- 7 vs. 36 +/- 7.5 years; P < 0.01). Many patients had vascular disease at more than one site: 17 had thrombophlebitis, 10 had arterial thromboses, and one had an aneurysm. Thrombophlebitis was more common in men (82% vs. 18%; P < 0.03) and arterial occlusion in women (70% vs. 30%; P > 0.05). Caval thrombosis and arterial occlusions were the most serious complications. Combined treatment with glucocorticoids, anticoagulants, and immunosuppressants was effective in superior vena cava syndrome and extracranial arterial occlusion. CONCLUSION: Vascular manifestations of BD are common in Lebanon, particularly venous lesions. Aneurysms are seen less often than arterial occlusions. Medical treatment may be sufficient in superior vena cava syndrome and arterial occlusion. PMID- 14563471 TI - Nicolau's syndrome after local glucocorticoid injection. AB - We report three cases of Nicolau's syndrome induced by intraarticular glucocorticoid injections. Nicolau's syndrome is defined as livedo-like dermatitis secondary to acute arterial thrombosis occurring immediately after intravascular injection of an insoluble drug substance. The cases described by Nicolau occurred in association with injections of oily bismuth suspensions. In 1970s and 1980s, cases occurred with delayed-action penicillin suspensions injected intramuscularly. Typically, the injection is followed immediately by excruciating pain in the buttock, sometimes with syncope. Cyanotic patches and a livedoid pattern develop. Rapid resolution of the pain and slower clearing of the skin changes occur in most patients. We report three typical cases with both severe pain and skin changes. In addition, two other patients had incomplete variants without skin abnormalities. Each of these five patients had received an injection in or about a joint of a glucocorticoid in a crystalline suspension. The pathophysiology of this syndrome probably involves acute vascular spasm related to penetration of microcrystals into a blood vessel. PMID- 14563472 TI - Intraosseous ganglion of the lunate: a case report. AB - An intraosseous ganglion of the lunate treated operatively, is reported. The patient suffered 2 years of pain in the left wrist and a cystic lesion in the lunate. Curettage and bone grafting resulted in complete relief of pain. PMID- 14563474 TI - Rheumatoid lung nodules with hydropneumothorax. PMID- 14563475 TI - Does infliximab decrease bone turnover in rheumatoid arthritis patients. PMID- 14563473 TI - Systemic sclerosis in a patient with recent nasopharyngeal carcinoma: an unusual combination. PMID- 14563476 TI - High-throughput, genome-scale protein production method based on the wheat germ cell-free expression system. AB - Cell-free protein synthesis systems can synthesize proteins with high speed and accuracy, but produce only a low yield because of their instability over time. Here we review our recent advances in a cell-free protein synthesis system prepared from wheat embryos. We first addressed and resolved the source of the instability of existing systems in light of endogenous ribosome-inactivating proteins. We found that conventional wheat germ extracts contained the RNA N glycosidase tritin and other inhibitors such as thionin, ribonucleases, deoxyribonucleases, and proteases that originate from the endosperm and inhibit translation. Extensive washing of wheat embryos to eliminate endosperm contaminants has resulted in extracts with a high degree of stability and activity. To maximize the translation yield and throughput of the system, we then focused on developing the following issues: optimization of the ORF flanking regions, a new strategy to construct PCR-generated DNAs for screening, and design of an expression vector for large-scale protein production. The resulting system achieves high-throughput expression, with a PCR-directed system at least 50 genes that can be translated in parallel, yielding between 0.1 and 2.3 mg of protein by one person within 2 days. Under the dialysis mode of reaction, the system with the expression vector can maintain productive translation for 14 days. The cell free system described here bypasses most of the biological processes and lends itself to robotic automation for high-throughput expression of genetic information, thus opening up many possibilities in the post-genome era. PMID- 14563477 TI - Chrysanthemum: advances in tissue culture, cryopreservation, postharvest technology, genetics and transgenic biotechnology. AB - Members of the Chrysanthemum-complex include important floricultural (cut-flower) and ornamental (pot and garden) crops, as well as plants of culinary, medicinal and (ethno)pharmacological interest. The last 35 years have seen a tremendous emphasis on their in vitro tissue culture and micropropagation, while the latter 10-15 years has seen a surge in transformation experiments, all aimed at ameliorating aesthetic and growth characteristics of the plants. This review highlights all available literature that exists on ornamental Chrysanthemum in vitro cell, tissue and organ culture, micropropagation and transformation. PMID- 14563478 TI - A unique central tryptophan hydroxylase isoform. AB - Serotonin (5-hydroxytryptophan, 5-HT) is a neurotransmitter synthesized in the raphe nuclei of the brain stem and involved in the central control of food intake, sleep, and mood. Accordingly, dysfunction of the serotonin system has been implicated in the pathogenesis of psychiatric diseases. At the same time, serotonin is a peripheral hormone produced mainly by enterochromaffin cells in the intestine and stored in platelets, where it is involved in vasoconstriction, haemostasis, and the control of immune responses. Moreover, serotonin is a precursor for melatonin and is therefore synthesized in high amounts in the pineal gland. Tryptophan hydroxylase (TPH) catalyzes the rate limiting step in 5 HT synthesis. Until recently, only one gene encoding TPH was described for vertebrates. By gene targeting, we functionally ablated this gene in mice. To our surprise, the resulting animals, although being deficient for serotonin in the periphery and in the pineal gland, exhibited close to normal levels of 5-HT in the brain stem. This led us to the detection of a second TPH gene in the genome of humans, mice, and rats, called TPH2. This gene is predominantly expressed in the brain stem, while the classical TPH gene, now called TPH1, is expressed in the gut, pineal gland, spleen, and thymus. These findings clarify puzzling data, which have been collected over the last decades about partially purified TPH proteins with different characteristics and justify a new concept of the serotonin system. In fact, there are two serotonin systems in vertebrates, independently regulated and with distinct functions. PMID- 14563479 TI - Novel roles for palmitoylation of Ras in IL-1 beta-induced nitric oxide release and caspase 3 activation in insulin-secreting beta cells. AB - We recently demonstrated that functional inactivation of H-Ras results in significant reduction in interleukin 1 beta (IL-1 beta)-mediated effects on isolated beta cells. Since palmitoylation of Ras has been implicated in its membrane targeting, we examined the contributory roles of palmitoylation of Ras in IL-1 beta-induced nitric oxide (NO) release and subsequent activation of caspases. Preincubation of HIT-T15 or INS-1 cells with cerulenin (CER, 134 microM; 3 hr), an inhibitor of protein palmitoylation, significantly reduced ( 95%) IL-1 beta-induced NO release from these cells. 2-Bromopalmitate, a structurally distinct inhibitor of protein palmitoylation, but not 2 hydroxymyristic acid, an inhibitor of protein myristoylation, also reduced (-67%) IL-1 beta-induced NO release from HIT cells. IL-induced inducible nitric oxide synthase gene expression was markedly attenuated by CER. Further, CER markedly reduced incorporation of [3H]palmitate into H-Ras and caused significant accumulation of Ras in the cytosolic fraction. CER-treatment also prevented IL-1 beta-induced activation of caspase 3 in these cells. Moreover, N-monomethyl-L arginine, a known inhibitor of inducible nitric oxide synthase, markedly inhibited IL-induced activation of caspase 3, thus establishing a link between IL induced NO release and caspase 3 activation. Depletion of membrane-bound cholesterol using methyl-beta-cyclodextrin, which also disrupts caveolar organization within the plasma membrane, abolished IL-1 beta-induced NO release suggesting that IL-1 beta-mediated Ras-dependent signaling in these cells involves the intermediacy of caveolae and their key constituents (e.g. caveolin 1) in isolated beta cells. Confocal light microscopic evidence indicated significant colocalization of Ras with caveolin-1. Taken together, our data provide the first evidence to indicate that palmitoylation of Ras is essential for IL-1 beta-induced cytotoxic effects on the islet beta cell. PMID- 14563480 TI - Induction of apoptosis without redox catastrophe by thioredoxin-inhibitory compounds. AB - The dithiol-reducing thioredoxin/thioredoxin reductase system normally maintains the reduced state of key enzymes responsible for the cell's anti-oxidant defences. We therefore addressed the question of whether AW 464--a novel thioredoxin inhibitor--as well as broad spectrum dithiol ligands diamide and phenylarsine oxide are able to induce and execute a regular apoptotic sequence of events without overwhelming the cell's ability to detoxify reactive oxygen species. All three agents were found to target the thioredoxin system in a cell free assay. In HL-60 leukaemia cells, they were also found to induce Bak activation, cytochrome c release from mitochondria, decreasing Delta Psi m, chromatin condensation, phosphatidyl serine exposure and Tdt-sensitive DNA nicks. At the onset of apoptosis there was no evidence of increases in oxygen free radicals or peroxide in cells treated with AW 464 or diamide. Phenylarsine oxide induced both free radicals and hydrogen peroxide, but this did not appear to interfere with apoptosis. We conclude that pharmacological targeting of thioredoxin can induce a well-orchestrated apoptotic programme. PMID- 14563481 TI - Antioxidant perturbations in the olfactory mucosa of alachlor-treated rats. AB - The chloracetanilide herbicide alachlor (2-chloro-2',6'-diethyl-N (methoxymethyl)acetanilide) induces olfactory mucosal tumors in rats following chronic dietary exposure. Previous reports demonstrated that alachlor exposure was associated with depletion of glutathione (GSH) in liver in vivo and in vitro, but did not address this issue in the target tissue for the carcinogenic response. In this study we investigated a potential oxidative stress pathway in olfactory tissue by examining perturbations in olfactory mucosal antioxidants. Male Long-Evans rats were fed alachlor for up to 10 days (10-126 mg/kg per day), and intracellular reduced GSH and ascorbate levels were measured in olfactory mucosa. Both GSH and ascorbate rapidly decreased in olfactory mucosa following alachlor exposure, with a subsequent increase in both antioxidants to approximately 160% of control levels in the high dose group, and recovery of GSH to control levels in all groups by 10 days. Using Western blot analysis, we found that the modifier subunit of the rate-limiting enzyme in GSH synthesis, glutamate cysteine ligase, increased in olfactory mucosa and remained elevated (126 mg/kg per day group). Two ascorbate transporters were detected by RT-PCR in olfactory mucosa, but neither appeared to be upregulated by alachlor exposure, and ascorbate synthesis was not stimulated in olfactory mucosa by alachlor treatment. Dietary exposure to alachlor depletes olfactory mucosa antioxidants, which may contribute to DNA damage and tissue-specific tumor formation. PMID- 14563482 TI - Spin trapping of superoxide, alkyl- and lipid-derived radicals with derivatives of the spin trap EPPN. AB - The N-t-butyl-alpha-phenylnitrone derivative N-2-(2-ethoxycarbonyl-propyl)-alpha phenylnitrone (EPPN) has recently been reported to form a superoxide spin adduct (t(1/2)=5.25 min at pH 7.0), which is considerably more stable than the respective N-t-butyl-alpha-phenylnitrone or 5,5-dimethylpyrroline N-oxide adducts (t(1/2) approximately 10 and 45s, respectively). In continuation of our previous studies on structure optimization of 5-(ethoxycarbonyl)-5-methyl-1-pyrroline N oxide derivatives, a series of six different EPPN derivatives was synthesized and characterized by 1H NMR, 13C NMR and IR spectroscopy. The ethoxy group of EPPN was replaced by a propoxy, iso-propoxy, n-butoxy, sec-butoxy, and tert-butoxy moiety, as well as the phenyl by a pyridyl ring. Electron spin resonance spectra and stabilities of the superoxide adducts of the propoxy derivatives were found to be similar to those of the respective EPPN adduct, whereas the electron spin resonance spectra of the superoxide adducts of N-2-(2-ethoxycarbonyl-propyl) alpha-(4-pyridyl) nitrone and the butoxy derivatives were accompanied by decomposition products. In contrast to the 5-(ethoxycarbonyl)-5-methyl-1 pyrroline N-oxide series, no significant improvement of the superoxide adduct stability could be obtained when the ethoxy group was replaced by other substituents. Carbon centered radical adducts derived from methanol, ethanol, formic acid and linoleic acid hydroperoxide were more stable than those of 5,5 dimethylpyrroline N-oxide, whereas among the alkoxyl radicals only the methoxyl radical adduct could be detected. PMID- 14563483 TI - Molecular mechanism of the enzymatic oxidation investigated for imidazoacridinone antitumor drug, C-1311. AB - The imidazoacridinone derivative, C-1311, is an antitumor agent that has been under phase I of clinical trial. The work presented here aims to elucidate the molecular mechanism of the enzymatic oxidative activation of this drug in such a model metabolic system, where the covalent binding to DNA was previously demonstrated. The oxidative activation of C-1311 was performed with HRP/H(2)O(2) and MPO/H(2)O(2) systems. The obtained final products of such transformations were separated and analysed by HPLC. The structures of the products were identified by means of ESI-MS and NMR. It was demonstrated that C-1311 was oxidised with HRP and MPO in the manner dependent on the drug:H(2)O(2) ratio and the drug was more susceptible to HRP oxidation than to MPO. Structural studies showed compounds C0 and C1 to be the result of dealkylation, which occurred in the amino groups of the side chain. The structures of C3 and C4 products were identified as dimers, whose monomers held the imidazoacridinone core. The activation of the imidazoacridinone ring system in position ortho to 8-hydroxyl group was necessary to form such dimers. We suggest that similar mechanism of C 1311 activation should occur in the presence of DNA when, instead of the dimer formation, the covalent binding to DNA, showed earlier for this drug, was formed. Since peroxidase-type enzymes are present in the cell nucleus of tumour cells the activation mechanisms of the C-1311 proposed here may be expected to take place in the cellular environment in vivo. PMID- 14563484 TI - Adenosine stimulation of the proliferation of colorectal carcinoma cell lines. Roles of cell density and adenosine metabolism. AB - Adenosine is a purine nucleoside which is present at micromolar concentrations in the extracellular fluid of solid cancers as a result of tissue hypoxia. Adenosine acts to promote tumor survival by inhibiting the cell-mediated anti-tumor immune response. However, its role in modulating proliferation of the tumor cell population is unclear. Differing results have been obtained using adenosine analogues or by interfering with adenosine metabolism. We examined the effect of adenosine itself on DNA synthesis and cell growth in six different human and mouse colorectal carcinoma cell lines, from different sites and at different stages of differentiation. Adenosine given as a single dose consistently stimulated DNA synthesis and cell proliferation in all cell lines tested, with an EC(50) of 3.8-30 microM and a maximum stimulation being reached at 10-100 microM. AMP and ATP also stimulated cell proliferation at similar doses. The stimulation by adenosine varied depending upon the culture cell density, with the greatest mitogenic effect at subconfluent densities. Adenosine was metabolized by cellular adenosine deaminase and adenosine kinase. The half-life (t(1/2)) for the decline in adenosine concentration in the medium following a single addition was between 40 min and 3 hr depending on the cell line and culture conditions. The rate of production of endogenous adenosine was low under normoxic culture conditions. Continuous dosing of cultures with adenosine to provide a steady-state concentration showed that proliferation could be stimulated by low micromolar concentrations of adenosine. We conclude that adenosine is stimulatory to the growth of human colorectal carcinoma cells at concentrations present within the tumor extracellular environment. PMID- 14563485 TI - L-Deprenyl as an inhibitor of menadione-induced permeability transition in liver mitochondria. AB - L-Deprenyl, an inhibitor of mitochondrial monoamine oxidase B (MAO B), inhibits the swelling of liver mitochondria induced by the pro-oxidant 2-methyl-1,4 naphtoquinone with a K(i) dependent on quinone concentration. L-Deprenyl also inhibits the collapse of membrane potential, cation efflux, pyridine nucleotide oxidation and cytochrome c release, all events which accompany the osmotic change and are typical of membrane permeability transition induction, thus emphasizing the inhibitory effect of the drug on this phenomenon. Results show that this inhibition is not due to the effect of L-deprenyl on monoamine oxidase activity but is most likely due to a direct interaction of the drug with the pore forming structures. It is here proposed that L-deprenyl, being a propargylamine, at physiological pH has a protonated amino group able to interact with critical aromatic or anionic amino acidic residues. As a consequence, the opening of the transition pore is prevented. These results indicate a more generalized protective effect of L-deprenyl on mitochondrial functions, involving the inhibition of membrane permeability transition induced not only by the oxidation of substrates of MAO B, but also by pro-oxidant agents such as 2-methyl-1,4 naphtoquinone, which does not involve MAO B activity. PMID- 14563487 TI - (-)-Epigallocatechin gallate causes oxidative damage to isolated and cellular DNA. AB - Green tea catechins, especially (-)-epigallocatechin gallate (EGCG), are believed to mediate much of the cancer chemopreventive effects of tea. However, it was reported that green tea catechins enhanced colon carcinogenesis in rats. Experiments using 32P-labeled DNA fragments obtained from human cancer-related genes showed that catechins induced DNA damage in the presence of metals such as Cu(II) and Fe(III) complexes. In the presence of Fe(III)EDTA, the order of DNA damaging ability was EGCG approximately (-)-epigallocatechin>(-)-epicatechin gallate>>catechin. Catechins plus Fe(III)EDTA caused DNA damage at every nucleotide, most likely due to *OH generation from H(2)O(2). In the presence of Cu(II), the order was (-)-epigallocatechin>catechin>EGCG>(-)-epicatechin gallate. Cu(II)-mediated DNA damage by EGCG occurred most frequently at T and G residues, especially of 5'-TG-3' and GG sequences. Catalase and bathocuproine inhibited the Cu(II)-mediated DNA damage, suggesting the involvement of H(2)O(2) and Cu(I). In the presence of metal ions, increased amounts of 8-oxo-7,8-dihydro-2' deoxyguanosine (8-oxodG) were found in DNA treated with EGCG. Furthermore, EGCG increased amounts of 8-oxodG in HL-60 cells, but not in the H(2)O(2)-resistant clone HP100. When GSH was reduced by L-buthionine-[S, R]-sulfoximine, a low concentration of EGCG increased amounts of 8-oxodG in HL-60 cells, further supporting the involvement of H(2)O(2) in cellular DNA damage. It is concluded that EGCG can induce H(2)O(2) generation and subsequent damage to isolated and cellular DNA, and that oxidative DNA damage may mediate the potential carcinogenicity of EGCG. PMID- 14563488 TI - Bcl-2 overexpression prevents daunorubicin-induced apoptosis through inhibition of XIAP and Akt degradation. AB - Daunorubicin (DNR) induces apoptosis in the human myeloid leukemia cells by activation of neutral sphingomyelinease and ceramide production. In the present study, we determined the effect of the antiapoptosis protein Bcl-2 on caspase-3 activation, phospholipase C-gamma 1 (PLC-gamma 1) degradation and cytochrome c release during the DNR-induced apoptosis. Treatment with 3 microM DNR for 12 hr produced morphological features of apoptosis and DNA fragmentation in U937 cells, which was associated with caspase-3 activation and PLC-gamma 1 degradation. Induction of apoptosis was also accompanied by release of cytochrome c, down regulation of X-linked inhibitor of apoptosis protein (XIAP), and inactivation of Akt, which was blocked by the pan-caspase inhibitor z-VAD-fmk. DNR-induced caspase-3 activation, PLC-gamma 1 degradation and apoptosis were significantly attenuated in Bcl-2 overexpressing U937/Bcl-2 cells. Ectopic expression of Bcl-2 appeared to inhibit DNR-induced apoptosis by interfering with inhibition of XIAP and Akt degradation. PMID- 14563486 TI - Identification and characterization of mechanistically distinct inducers of gamma globin transcription. AB - Inhibition of HbS polymerization is a major target for therapeutic approaches in sickle cell anemia. Toward this goal, initial efforts at pharmacological elevation of fetal hemoglobin (HbF) has shown therapeutic efficacy. In order to identify well-tolerated, novel agents that induce HbF in patients, we developed a high-throughput screening approach based on induction of gamma-globin gene expression in erythroid cells. We measured gamma-globin transcription in K562 cells transfected with either gamma promoter elements fused with the locus control region hypersensitivity site 2 and luciferase reporter gene (HS2 gamma) or a beta-yeast artificial chromosome in which the luciferase reporter gene was recombined into the gamma-globin coding sequences (gamma YAC). Corresponding pharmacological increases in HbF protein were confirmed in both K562 cells and in human primary erythroid progenitor cells. Approximately 186,000 defined chemicals and fungal extracts were evaluated for their ability to increase gamma gene transcription in either HS2 gamma or gamma YAC models. Eleven distinct classes of compounds were identified, the majority of which were active within 24-48 hr. The short chain hydroxamate-containing class generally exhibited delayed maximal activity, which continued to increase transcription up to 120 hr. The cyclic tetrapeptide OSI-2040 and the hydroxamates were shown to have histone deacetylase inhibitory activity. In primary hematopoietic progenitor cell cultures, OSI-2040 increased HbF by 4.5-fold at a concentration of only 40 nM, comparable to the effects of hydroxyurea at 100 microM. This screening methodology successfully identifies active compounds for further mechanistic and preclinical evaluation as potential therapeutic agents for sickle cell anemia. PMID- 14563489 TI - Aggravation of L-DOPA-induced neurotoxicity by tetrahydropapaveroline in PC12 cells. AB - Tetrahydropapaveroline (THP) is formed in Parkinsonian patients receiving L-DOPA therapy and is detected in the plasma and urine of these patients. In this study, we have investigated the effects of THP on L-DOPA-induced neurotoxicity in cultured rat adrenal pheochromocytoma, PC12 cells. Exposure of PC12 cells up to 10 microM THP or 20 microM L-DOPA after 24 or 48 hr, neither affected the cell viability determined by MTT assay, nor induced apoptosis by flow cytometry and TUNEL staining. However, at concentrations higher than 15 microM, THP showed cytotoxicity through an apoptotic process. In addition, THP at 5-15 microM for both incubation time points significantly enhanced L-DOPA-induced neurotoxicity (L-DOPA concentration, 50 microM). Exposure of PC12 cells to THP, L-DOPA and THP plus L-DOPA for 48 hr resulted in a marked increase in the cell loss and percentage of apoptotic cells compared with exposure for 24hr. The enhancing effects of THP on L-DOPA-induced neurotoxicity were concentration- and treated time-dependent. THP, L-DOPA and THP plus L-DOPA produced a significant increase in intracellular reactive oxygen species generation and decrease in ATP levels, supporting the involvement of oxidative stress in THP- and L-DOPA-induced apoptosis. The antioxidant N-acetyl-L-cysteine strongly inhibited changes in apoptosis, decreases in cell viability and ROS generation induced by THP associated with L-DOPA. These results suggest that THP aggravates L-DOPA-induced oxidative neurotoxic and apoptotic effects in PC12 cells. Therefore, Parkinsonian patients treated with L-DOPA for long-term need to be monitored for the relationship between plasma concentration of THP and the symptoms of neurotoxicity. PMID- 14563490 TI - Induction by staurosporine of hepatocyte growth factor production in human skin fibroblasts independent of protein kinase inhibition. AB - Staurosporine is one of the most potent and well known inhibitors of protein kinases, and it is often used to study the involvement of protein kinases in signal transduction pathways. We now report that staurosporine can induce the production of hepatocyte growth factor (HGF) independently of protein kinase inhibition. Staurosporine markedly stimulated the production of HGF in various cell types, including human skin fibroblasts. Its effect was accompanied by up regulation of HGF gene expression. The inhibition of protein kinases appears not to be involved in staurosporine-induced HGF production, because other protein kinase inhibitors, K-252a, H-7, GF 109203X and genistein, had no HGF-inducing activity. UCN-01, 7-hydroxystaurosporine, which differs from staurosporine only in its aglycone moiety, also showed HGF-inducing activity, and inactive K-252a differs from staurosporine only in its sugar moiety. These results indicate that the sugar moiety, a six-atom ring structure, is important in the HGF-inducing activity of staurosporine. Experiments were then carried out to determine whether the characteristics of staurosporine-induced HGF production have similarities to those of HGF production stimulated by other HGF inducers. The effect of staurosporine like that of 8-bromo-cAMP and that of cholera toxin was marked in human skin fibroblasts from all four different sources, whereas the effects of epidermal growth factor and phorbol 12-myristate 13-acetate were variable depending on cells. The net increase in HGF production induced by staurosporine was not reduced in protein kinase C-depleted human skin fibroblasts. Moreover, synergistic induction of HGF was detected between staurosporine and interferon gamma as well as between 8-bromo-cAMP and interferon-gamma. Staurosporine, however, did not increase intracellular cAMP levels in human skin fibroblasts. These results indicate that staurosporine induced HGF in different cell types via a signaling pathway similar to the cAMP-mediated pathway without increasing cAMP levels. PMID- 14563491 TI - Chemically modified tetracyclines selectively inhibit IL-6 expression in osteoblasts by decreasing mRNA stability. AB - In bone biology, interleukin (IL)-6 is an autocrine/paracrine cytokine which can induce osteoclasts formation and activation to help mediate inflammatory bone destruction. Previous studies have shown that tetracycline and its derivatives have potentially beneficial therapeutic effects in the prevention and treatment of metabolic bone diseases by modulating osteoblast and osteoclast activities. Our previous studies indicated that non-antimicrobial chemically modified tetracyclines (CMTs) can dose-dependently inhibit IL-1 beta-induced IL-6 secretion in osteoblastic cells. In the present study, we explored the molecular mechanisms underlying the ability of doxycycline analogs CMT-8 and its non chelating pyrazole derivative, CMT-5 to affect IL-6 gene expression in murine osteoblasts. Steady-state IL-6 mRNA was decreased with CMT-8 (ca. 50%) but not by CMT-5 when stimulated by IL-1 beta. CMT-8 regulation of IL-1 beta-induced IL-6 gene expression was further explored. CMT-8 did not affect IL-6 promoter activity in reporter gene assays. However, the IL-6 mRNA stability was decreased in the presence of CMT-8. These effects require de novo protein synthesis as they were inhibited by cycloheximide. Western blot analysis indicated that CMT-8 did not affect p38 mitogen-activated protein kinase, c-jun NH(2)-terminal kinases, or extracellular signal-regulated kinases (1 and 2) phosphorylation in response to IL-1 beta. These data suggest that CMT-8 can modulate inhibit IL-1 beta-induced IL-6 expression in MC3T3-E1 cells at the post-transcriptional level affecting IL 6 mRNA stability. These observations may offer a novel molecular basis for this treatment of metabolic bone diseases that are mediated by IL-6. PMID- 14563492 TI - Inhibition of lipopolysaccharide-induced nitric oxide production by flavonoids in RAW264.7 macrophages involves heme oxygenase-1. AB - The role of heme oxygenase-1 (HO-1) played in the inhibitory mechanism of flavonoids in lipopolysaccharide (LPS)-induced responses remained unresolved. In the present study, flavonoids, including 3-OH flavone, baicalein, kaempferol, and quercetin, induced HO-1 gene expression at the protein and mRNA levels in the presence or absence of LPS in RAW264.7 macrophages. This effect was associated with suppression of LPS-induced nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) protein expression. Hemin induced HO-1 protein expression and this was associated with the suppression of LPS-induced NO production and iNOS protein expression in a dose-dependent manner. In addition, an increase in bilirubin production was found in flavonoid- and hemin-treated cells. Hemin, at the doses of 10, 20, and 50 microM, dose-dependently stimulated the flavonoid (50 microM)-induced HO-1 protein expression, and enhanced their inhibitory effects on LPS-induced NO production and iNOS protein expression. Pretreatment of the HO-1 inhibitor, tin protoporphyrin (10 microM), attenuated the inhibitory activities of the indicated flavonoids on LPS-induced NO production. Morphologic analysis showed that 3-OH flavone, baicalein, kaempferol, quercetin, hemin, and tin protoporphyrin did not cause any change in cell viability in the presence or absence of LPS. In contrast, only 3-OH flavone showed a significant inhibition of cell growth using the MTT assay. Transfection of an HO-1 vector in macrophages (HO-1/RAW264.7) resulted in a 3-fold increase in HO-1 protein compared with that the parental RAW264.7 cells. NO production mediated by LPS in HO-1 over-expressed RAW264.7 cells (HO-1/RAW264.7) was significant less than that in parental RAW264.7 cells. 3-OH Flavone, baicalein, kaempferol, and quercetin showed a more significant inhibition on LPS-induced NO production in HO-1/RAW264.7 cells than in parental RAW264.7 cells. These results provide evidence on the role of HO-1 in the inhibition of LPS-induced NO production by flavonoids. A combination of HO-1 inducers (i.e. hemin) and flavonoids might be an effective strategy for the suppression of LPS-induced NO production. PMID- 14563493 TI - Characterisation of C5a receptor agonists from phage display libraries. AB - C5a des-Arg(74) has a 10- to 100-fold lower receptor binding affinity than intact C5a and is only a partial agonist. We have used phage display selection from randomly mutated C5a des-Arg(74) libraries to isolate variant proteins that can activate C5a receptors with similar potency to C5a. Here we explore the interactions of three variants (V1-3) with C5aR mutated at residues involved in the differential response. The mutant Asp(282)Arg-C5aR is preferentially activated by C5a des-Arg(74), probably due to repulsion between Arg(74) of C5a and the substituent Arg(282). In accordance with this hypothesis, V2 (with a polar C-terminus which has no Arg residue) but not V1 (with a C-terminal Arg residue at position 73) could activate Asp(282)Arg-C5aR. V3, with a very hydrophobic C-terminus, was the most potent agonist at Asp(282)Arg-C5aR. Arg(175) is a potential counterion for the C-terminal carboxylate of C5a. C5aR mutated to either Ala or Asp at this position lost nearly all responsiveness to both C5a and C5a des-Arg(74), suggesting that mutation of Arg(175) caused a non-specific loss of receptor conformation and a loss of signalling capacity. However, V3 could still activate Arg(175)Asp/Ala-C5aR with the same potency as wild-type C5aR, demonstrating that the mutant receptors retained high signalling capability and showed a specific loss of responsiveness. Thus C5a des-Arg(74) variants produced by phage display are potentially useful tools for the dissection of ligand receptor interactions. PMID- 14563494 TI - Novel chemoattractant peptides for human leukocytes. AB - Phospholipase A(2) plays a key role in phagocytic cell functions. By screening a synthetic hexapeptide combinatorial library, we identified 24 novel peptides based on their ability to stimulate arachidonic acid release associated with cytosolic phospholipase A(2) activity in differentiated HL60 cells. The identified peptides, that contain the consensus sequence (K/R/M)KYY(P/V/Y)M, also induce intracellular calcium release in a pertussis toxin-sensitive manner showing specific action on phagocytic leukocytes, but not on other cells. Functionally, the peptides stimulate superoxide generation and chemotactic migration in human neutrophils and monocytes. Four of the tested active peptides were ligands for formyl peptide receptor like 1. Among these, two peptides with the consensus sequence (R/M)KYYYM can induce intracellular calcium release in undifferentiated HL60 cells that do not express formyl peptide receptor like 1, indicating usage of other receptor(s). A study of intracellular signaling in differentiated HL60 cells induced by the peptides has revealed that four of the novel peptides can induce extracellular signal-regulated protein kinase activation via shared and distinct signaling pathways, based on their dependence of phospatidylinositol-3-kinase, protein kinase C, and MEK. These peptides provide previously unavailable tools for study of differential signaling in leukocytes. PMID- 14563495 TI - Fangchinoline inhibits rat aortic vascular smooth muscle cell proliferation and cell cycle progression through inhibition of ERK1/2 activation and c-fos expression. AB - Fangchinoline (FAN; a plant alkaloid isolated from Stephania tetrandrae) is a nonspecific Ca(2+) channel blocker. The objective of the present study was to investigate the effect of FAN on the growth factor-induced proliferation of primary cultured rat aortic smooth muscle cells (RASMCs). FAN significantly inhibited both 5% fetal bovine serum (FBS)- and 50ng/mL platelet-derived growth factor (PDGF)-BB-induced proliferation, [3H]thymidine incorporation into DNA and phosphorylation of extracellular signal-regulated kinase 1/2. In accordance with these findings, FAN revealed blocking of the FBS-inducible progression through G(0)/G(1) to S phase of the cell cycle in synchronized cells and caused a 62% decrease in the early elevation of c-fos expression induced after 5% FBS addition. Furthermore, significant antiproliferative activity of FAN is observed at concentrations below those required to achieve significant inhibition of Ca(2+) channels by FAN. These results suggest that FAN reduced both FBS- and PDGF BB-induced RASMCs proliferation by perturbing cell cycle progression. This antiproliferative effect of FAN is dependent on the MAP kinase pathway, but cannot be limited to its Ca(2+) modulation. PMID- 14563496 TI - Sphingosine 1-phosphate induces CREB activation in rat cerebral artery via a protein kinase C-mediated inhibition of voltage-gated K+ channels. AB - Sphingosine 1-phosphate (S1P) is a potential mitogenic stimulus for vascular smooth muscle. S1P promotes an increase in the intracellular calcium concentration ([Ca(2+)](i)) in cerebral arteries, however S1P effects on regulation of gene expression are not known. Activation of the Ca(2+)-dependent transcription factor, cAMP response element-binding protein (CREB), is associated with smooth muscle proliferation. The aim of this study was to examine the Ca(2+) dependent mechanisms involved in S1P-induced CREB activation in cerebral artery. Western blotting and immunofluorescence with a phospho-CREB antibody were used to detect CREB activation in Sprague-Dawley rat cerebral arteries. Whole-cell patch clamp recording and single cell imaging of [Ca(2+)](i) were performed on freshly isolated cerebral artery myocytes. S1P increased activation of CREB in the nucleus of cerebral arteries. This activation was mediated by Ca(2+)/calmodulin dependent protein kinase and was dependent on an increase in [Ca(2+)](i) via two mechanisms: (i) intracellular Ca(2+) release via an inositol 1,4,5-trisphosphate (InsP(3))-dependent pathway and (ii) Ca(2+) entry through voltage-dependent Ca(2+) channels (VDCC). Activation of the VDCC occurred through S1P-induced inhibition (approximately 50%) of the voltage-gated potassium (K(+)) current. This inhibition was via a protein kinase C-mediated pathway resulting in tyrosine phosphorylation of at least one isoform of the Kv channel (Kv 1.2). These results demonstrate that S1P can activate the transcription factor CREB through different Ca(2+)-dependent pathways including intracellular Ca(2+) release and inhibition of voltage-gated K(+) channels leading to Ca(2+) influx. Our findings suggest a potential role for S1P in regulation of gene expression in vascular smooth muscle. PMID- 14563497 TI - Influence of lidocaine on ouabain-induced inotropic response in rat atria. AB - In this paper we demonstrated that lidocaine broadens the therapeutic range of ouabain action having a protective effect on ouabain-induced toxicity on rat atria. The lidocaine effect on therapeutic ouabain action was associated with the increase in the sensitivity of Na(+)-K(+)-ATPase related to a decreased in the equilibrium dissociation constant (K(d)) of high affinity binding sites. Lidocaine suppressed the ouabain-induced tonotropic effect and arrhythmias, decreasing the number of low affinity binding sites (B(max)) without changes in K(d). Blockade of Na(+)-Ca(2+) exchange with KB-R7943 or dual Na(+)-Ca(2+) channel with flunarizine, mimicked lidocaine effect increasing ouabain therapeutic action, extending its concentration range tolerated, delaying the onset of contracture. Lidocaine itself triggered negative inotropic response at high concentration. This effect was increased in the presence of flunarizine and verapamil but not by the inhibition of calcium/calmodulin with W-7. The mechanism underlying the lidocaine-induced negative inotropic response, appears to be different that underlying the positive inotropic effect on ouabain action. This study provides evidence that lidocaine can interact with the same or similar binding sites for ouabain in rat atrial tissue, providing a protective effect on ouabain-induced changes in contractility. The contribution of Na(+)-Ca(2+) exchange and/or Ca(2+) overload on lidocaine effect is discussed. PMID- 14563498 TI - Cytokines, insulin-like growth factor 1, sarcopenia, and mortality in very old community-dwelling men and women: the Framingham Heart Study. AB - BACKGROUND: Aging is associated with increased production of catabolic cytokines, reduced circulating levels of insulin-like growth factor 1 (IGF-1), and acceleration of sarcopenia (loss of muscle with age). We hypothesized that these factors are independently linked to mortality in community-dwelling older persons. METHODS: We examined the relation of all-cause mortality to peripheral blood mononuclear cell production of inflammatory cytokines (tumor necrosis factor alpha [TNF-alpha], interleukin 1 beta, interleukin 6), serum interleukin 6 and IGF-1, and fat-free mass and clinical status in 525 ambulatory, free-living participants in the Framingham Heart Study. RESULTS: Of the 525 subjects (aged 72 to 92 years at baseline), 122 (23%) died during 4 years of follow-up. After adjusting for age, sex, comorbid conditions, smoking, and body mass index, mortality was associated with greater cellular production of TNF-alpha (hazard ratio [HR] = 1.27 per log(10) difference in ng/mL; 95% confidence interval [CI]: 1.00 to 1.61; P = 0.05) and higher serum interleukin 6 levels (HR = 1.30 per log(10) difference in pg/mL; 95% CI: 1.04 to 1.63]; P = 0.02), but not with higher serum IGF-1 levels (HR = 0.70 per log(10) difference in pg/mL; 95% CI: 0.49 to 0.99; P = 0.04). In a subset of 398 subjects (55 deaths) in whom change in fat-free mass index during the first 2 years was measured, less loss of fat free mass and greater IGF-1 levels were associated with reduced mortality during the next 2 years. CONCLUSION: Greater levels or production of the catabolic cytokines TNF-alpha and interleukin 6 are associated with increased mortality in community-dwelling elderly adults, whereas IGF-1 levels had the opposite effect. PMID- 14563499 TI - Comparison of four clinical prediction scores for the diagnosis of lower limb deep venous thrombosis in outpatients. AB - PURPOSE: We compared three scores for the prediction of deep venous thrombosis with a new score designed specifically for outpatients. METHODS: Patients referred for evaluation because of suspected deep venous thrombosis were examined by ultrasonography. Sensitivity and specificity were calculated for three clinical scores (Wells [nine components], Kahn [four components], and St. Andre [six components]). We developed a new score by multivariate analysis, and then compared this score with the others in a new sample. RESULTS: Four hundred and forty-four outpatients were included in the first sample, of whom 126 (28%) had deep venous thrombosis. The Wells score was a better predictor of deep venous thrombosis than the Kahn and St. Andre scores. According to the Wells score, 73 patients had a high probability of deep venous thrombosis (of whom 51 [70%] actually had a thrombosis) and 178 had a low probability of deep venous thrombosis (of whom 19 [11%] had a thrombosis). A new score was developed as follows: male sex (+1), lower limb palsy or immobilization (+1), confinement to bed >3 days (+1), lower limb enlargement (+1), unilateral lower limb pain (+1), and other plausible diagnosis (-1). In a validation sample of 282 outpatients, this score identified 31 patients who had a high probability of deep venous thrombosis (score > or =3), of whom 18 (58%) had a thrombosis, and 70 patients who had a low probability (score < or =0), of whom 3 (4%) had a thrombosis. The Wells score and this ambulatory score had similar test operating characteristics in the validation sample. CONCLUSION: Our new six-component score had similar diagnostic utility as the nine-component Wells score among outpatients being evaluated for deep venous thrombosis. PMID- 14563500 TI - A randomized controlled trial of a Chinese herbal remedy to increase energy, memory, sexual function, and quality of life in elderly adults in Beijing, China. AB - BACKGROUND: Chinese herbal medicines are commonly used to improve general health and well-being despite limited scientific data to support their efficacy. We conducted a randomized, double-blind, placebo-controlled trial to determine whether an herbal remedy that is used widely in China was associated with changes in quality of life, energy, memory, sexual function, and qi (the Chinese concept of "vital energy" that is important in general health). METHODS: Residents (n = 237) of Beijing, China, who were aged > or =60 years and had self-reported decreased energy, memory, or sexual function, were randomly assigned to take four tablets of a Chinese herbal formula or of an identical placebo, three times a day for 30 days. Patients returned for one follow-up visit after 30 days for assessment of all outcomes. The main outcome measures were changes in quality of life at 30 days as measured by the 12-Item Short Form Health Survey (SF-12) Mental and Physical Component Summary scales. RESULTS: Use of Chinese herbs was associated with a small benefit in the Mental Component scale (difference of 1.9 points; 95% confidence interval [CI]: 0.1 to 3.6) and no benefit in the Physical Component scale (difference of -0.1 points; 95% CI: -1.7 to 1.5) as compared with placebo. A small improvement in the qi scale was no longer significant after adjusting for baseline differences in this score between groups. There was no improvement in physical performance, memory, or sexual function. The herbal product was well tolerated. CONCLUSION: Short-term use of a mixture of Chinese herbs was associated with a small benefit in one measure of mental health that is of unclear clinical importance. PMID- 14563501 TI - Is geography destiny for patients in New York with myocardial infarction? AB - PURPOSE: The use of coronary revascularization among patients with myocardial infarction varies by race/ethnicity and socioeconomic status. The objective of this study was to determine whether local availability of facilities might influence apparent racial disparities in revascularization and health outcomes. METHODS: Using Statewide Planning and Research Cooperative System data (1988 1999) from the New York State Department of Health, we determined revascularization rates among patients hospitalized with myocardial infarction in two socioeconomically disadvantaged communities in New York City (the South Bronx, which has no hospitals that have revascularization facilities, and Harlem, which has three revascularization facilities), as well as in its most advantaged community (mid-Manhattan, which has six such facilities). The rest of New York City served as reference. We measured demographic and clinical characteristics and revascularization rates in each community. RESULTS: Among patients hospitalized with myocardial infarction, the age-adjusted revascularization rates were 29.2% for whites, 12.5% for blacks, and 19.9% for Hispanics (P <0.01). Rates were 12.0% in the South Bronx, 24.0% in Harlem, 38.4% in mid-Manhattan, and 21.2% in the rest of New York City (P <0.05). Logistic regression analysis, adjusting for age, sex, race, insurance status, comorbidity, clinical complications, and year of admission, revealed that South Bronx patients were about 20% less likely to be revascularized than those in the rest of New York City, whereas patients living in Harlem were twice as likely to receive such treatment as residents in the rest of New York City. Among patients admitted to hospitals with cardiac revascularization facilities, lower use among South Bronx residents persisted, but after adjusting for patient characteristics, Harlem residents were significantly less likely to be revascularized than those from the rest of New York City. CONCLUSION: Race and socioeconomic factors influence the likelihood of revascularization after myocardial infarction among residents of New York City. In addition, lack of availability of revascularization further reduces its use by residents of disadvantaged neighborhoods. PMID- 14563502 TI - Effects of previous influenza vaccination on subsequent readmission and mortality in elderly patients hospitalized with pneumonia. AB - PURPOSE: To determine the effect of influenza vaccination on mortality and hospital readmission rates following discharge of elderly patients admitted with pneumonia. METHODS: We reviewed the medical records of 12,566 randomly selected Medicare beneficiaries hospitalized for pneumonia from October 1 through December 31, 1998, to assess mortality and hospital readmission rates from the date of discharge through the influenza season, May 1, 1999. Patients were grouped based on vaccination status: before hospitalization, during hospitalization, or unknown (no evidence of vaccination). RESULTS: Severity-adjusted mortality rates were 22.4% (95% confidence interval [CI]: 14.4% to 29.7%) for the vaccination before hospitalization group, 26.4% (95% CI: 20.4% to 31.9%) for the in-hospital vaccination group, and 29.4% (95% CI: 28.1% to 30.6%) for the unknown vaccination status group. Patients vaccinated before hospitalization had significantly lower mortality than did patients with unknown vaccination status (hazard ratio [HR] = 0.65; 95% CI: 0.59 to 0.70; P <0.0001). Adjusted readmission rates were 42.6% (95% CI: 40.0% to 45.1%) for the vaccination before hospitalization group, 40.0% (95% CI: 33.2% to 46.1%) for the in-hospital vaccination group, and 44.8% (95% CI: 43.3% to 46.4%) for the unknown vaccination status group. Patients vaccinated before hospitalization had significantly lower readmission rates than patients with unknown vaccination status (HR = 0.92; 95% CI: 0.87 to 0.98; P = 0.009). CONCLUSION: Influenza vaccination before hospitalization was effective in decreasing subsequent mortality and hospital readmission in elderly patients with pneumonia. PMID- 14563503 TI - Early and late effects of low-dose aspirin on renal function in elderly patients. AB - BACKGROUND: Although low-dose aspirin is used by many elderly patients, monitoring of renal function is currently not recommended. We recently reported transient retention of uric acid and creatinine caused by aspirin in doses of 75 to 325 mg/d. We therefore evaluated the renal effects of aspirin (100 mg/d), including post-treatment effects. METHODS: We studied 83 stable geriatric patients in long-term care (aged 56 to 98 years) who were treated with low-dose aspirin (100 mg/d) for 2 weeks and 40 control patients. Other medications and diet were kept constant. Biochemical monitoring including blood samples and 24 hour urinary collections for creatinine and uric acid at baseline and weekly for a total of 5 weeks. RESULTS: After 2 weeks on aspirin, urinary excretion of creatinine decreased in 60 (72%) and excretion of uric acid decreased in 54 (65%) of the 83 patients, and their mean clearances decreased; during the same period, serum blood urea nitrogen, creatinine, and uric acid levels increased (P <0.05 for all). Deterioration from baseline levels was significantly greater (and more prevalent) in the aspirin-treated group than in the 40 control patients (P = 0.001 to 0.09). After withdrawal of aspirin these parameters improved. However, 3 weeks after stopping aspirin, 48% (35 of the 73 in whom this measurement was available) had a persistent decline in creatinine clearance from baseline, as compared with only 8% (3/36) controls (P <0.001). CONCLUSION: Short-term low-dose aspirin treatment may affect renal function in elderly patients. These effects persist 3 weeks after cessation of the drug in some of these patients. PMID- 14563504 TI - Effects of early and late administration of angiotensin-converting enzyme inhibitors on mortality after myocardial infarction. AB - BACKGROUND: Clinical trials demonstrating that angiotensin-converting enzyme (ACE) inhibitors reduce mortality when prescribed following myocardial infarction have focused on either early or late administration. It is unclear what the overall mortality benefits are and how long they last. METHODS: We identified trials of ACE inhibitor therapy after myocardial infarction that were published between 1987 and 2002. Studies were included if they were randomized placebo controlled trials with follow-up of at least 1 month. Separate analyses were performed for trials of therapy initiated within 48 hours of infarction (early administration) or initiated more than 48 hours after infarction (late administration). The Yusuf-Peto modification of the Mantel-Haenszel method was used to obtain summary odds ratios for mortality. RESULTS: Twenty-two trials (14 early administration, 8 late administration) met the inclusion criteria. Early administration of ACE inhibitors was associated with a slight reduction in 1 month mortality (odds ratio [OR] = 0.93; 95% confidence interval [CI]: 0.88 to 0.97) and a much larger reduction in 1-year mortality (OR = 0.68; 95% CI: 0.54 to 0.87). Late therapy was associated with significant reductions in 1-year mortality (OR = 0.84; 95% CI: 0.73 to 0.97), 2-year mortality (OR = 0.75; 95% CI: 0.67 to 0.85), and 3-year mortality (OR = 0.74; 95% CI: 0.65 to 0.84). CONCLUSION: Both early and late administration of ACE inhibitor therapy are associated with lower mortality following myocardial infarction, with the largest benefits observed with long-term mortality. PMID- 14563505 TI - Update on therapy for heart failure. AB - The success of angiotensin-converting enzyme (ACE) inhibitors in reducing morbidity and mortality in patients with heart failure has led to investigations of other inhibitors of the renin-angiotensin-aldosterone system. Although ACE inhibitors remain first-line drugs in the treatment of heart failure and left ventricular dysfunction, clinical evidence suggests that a newer class of agents- angiotensin II receptor blockers--may provide additional benefit by blocking the adverse effects of angiotensin II more completely. An improved adverse-effect profile also makes angiotensin II receptor blockers appropriate in patients who cannot tolerate ACE inhibitors. Clinical trials have demonstrated the beneficial effects of angiotensin II receptor blockers on the combined endpoints of morbidity and mortality in patients with heart failure. Aldosterone antagonism with spironolactone has additive benefits in patients receiving an ACE inhibitor. The most recent treatment guidelines for heart failure recommend the use of angiotensin II receptor blockers and spironolactone in selected patients. PMID- 14563506 TI - Mycotic aneurysms of the thoracic aorta: a diagnostic challenge. PMID- 14563507 TI - Cases from the Osler Medical Service at Johns Hopkins University. PMID- 14563508 TI - Inflammation, hormones, and body composition at a crossroad. PMID- 14563509 TI - Early and late angiotensin-converting enzyme inhibition after myocardial infarction: an overview of randomized clinical trials. PMID- 14563510 TI - Atypical manifestations of infections in patients with familial dysautonomia. PMID- 14563512 TI - Early occurrence of septic pericarditis after Nissen fundoplication. PMID- 14563511 TI - Acute renal failure due to the herbal remedy CKLS. PMID- 14563515 TI - Aging and the hemopoietic system. AB - While increasing numbers of elderly patients are expected to require chemotherapy and/or radiotherapy in the future, the application of standard-dose chemotherapy in unselected cohorts of older patients usually results in a higher rate of life threatening myelosuppression and treatment-related deaths compared to young individuals treated with the same chemotherapy regimens. The biologic mechanisms underlying reduced tolerance to chemotherapy of the hemopoietic system in older individuals are still poorly understood. Unveiling such mechanisms therefore represents a fundamental issue to ameliorate chemotherapy strategies for older cancer patients. Current evidence suggests that aging-related bone marrow changes are rather subtle and most probably irrelevant for the hemopoietic function of normal older individuals. These changes, however, may become clinically evident under conditions of severe hemopoietic stress such as the administration of repeated courses of chemo-radiotherapy. The mechanisms underlying age-dependent decline in the hematopoietic reserve are not fully clarified and probably involve age-associated changes in the stem and progenitor cells compartments which may ultimately lead to a reduced ability of recovery from hematologic stress. Age related changes in endogenous anti-tumor immune responses also need to be accounted for in the aim of managing residual disease in elderly cancer patients treated with effective chemo-radiotherapy. Since dendritic cells (DCs) generated from older individuals appear fully functional, dendritic cell-based immunotherapy may represent an important tool to treat residual disease in aged cancer patients. While it is clearly established that primary prophylaxis with hemopoietic growth factors currently enables a large fraction of older cancer patients to receive appropriate chemotherapy, innovative strategies in the use of such growth factors may allow time-intensification of standard-dose chemotherapy for treating chemosensitive tumors, i.e. non-Hodgkin's lymphomas (NHLs), occurring in older individuals. PMID- 14563516 TI - Thrombocytopenia and hemorrhagic risk in cancer patients. AB - In patients with malignant diseases, despite a direct relationship between the level of platelet count and bleeding episodes, it is evident that the risk of bleeding depends not only on the platelet count, but also on the underlying disease, the use of drugs interfering with platelet function and complications such as fever and infection or the presence of coagulation defects. As a consequence, it is not only the absolute platelet count, but rather the number of functional platelets that is important for the prevention of bleeding. As for Prophylaxis of hemorrhagic risk in these setting of patients, studies have now convincingly demonstrated that a 10,000/microL threshold for prophylactic platelet transfusion is safe and effective in uncomplicated thrombocytopenic patients. However, a threshold of 20,000/microL must be considered only for patients with high fever, hyperleukocytosis, coagulation abnormalities or who necessitate of invasive procedure. Moreover, as suggested by the clinical practice guidelines of the American Society of Clinical Oncology, a threshold of 20,000/microL must be considered for patients receiving aggressive therapy for bladder tumors and for those with demonstrated necrotic tumors. PMID- 14563517 TI - Acute myeloid leukemia in the older patient. AB - Acute myeloid leukemia (AML) is an extremely heterogeneous disorder. The biology of AML is incompletely understood, but much data indicates that older patients have a more biologically diverse and chemotherapy resistant form of AML that is quite different from that seen in the younger patients. Approximately 60% of AML cases are in patients greater than 60 years of age, so the predominant burden is in older patients. This problem will be magnified in the future, because the US population is both growing and aging. When one examines the treatment outcomes of older AML patients over the last three decades, there is little progress in long term survival. Nine major published randomized placebo controlled trials of myeloid growth factors given during induction for AML have been conducted. All of these trials with one exception demonstrated no significant impact on the clinical outcomes of complete response (CR) rate, disease-free, and overall survival. However, the duration of neutropenia was consistently and uniformly reduced by the use of growth factor in all nine of these trials. Because of the favorable impact of the colony-stimulating factors (CSFs) on resource use, antibiotic days, hospital days, etc., it can be more economical and beneficial to use CSFs in AML than to withhold use. The overall dismal outlook for the older AML patient can only be altered by clinical trials with new therapeutic agents. New cellular and molecularly targeted agents are entering clinical trials and bring hope for progress to this area of cancer therapy. PMID- 14563518 TI - Complications of cytotoxic chemotherapy in older patients: focus on myelotoxicity in lymphomas. AB - The management of the older lymphoma patient with cytotoxic chemotherapy requires knowledge of the pharmacologic effects of age and awareness of the diversity of the older population. The most obvious reason for attenuation of treatment intensity in the elderly is the possibility of a decline in hemopoietic reserve. The research of specific therapeutic protocols is needed to cure the elderly lymphoma patients reducing the risk of important toxic effects. PMID- 14563519 TI - The use of colony stimulating factors in elderly patients with cancer. AB - Hematological toxicity is the most common and the most frequent fatal complication of chemotherapy. It is observed with increased frequency with age, it is a significant independent predictor of the development of febrile neutropenia, and may contribute to a reluctance to administer chemotherapy in the elderly patient population. The authors analyze published data on effectiveness and results of the use of colony stimulating factors for preventing and treating elderly patients affected by tumors during chemotherapy. PMID- 14563520 TI - Optimising management of neutropenia and anaemia in cancer chemotherapy-advances in cytokine therapy. AB - Neutropenia and anaemia are serious complications of myelosuppressive chemotherapy. They have a negative impact on patient quality of life and may reduce response to treatment. Febrile neutropenia, a potentially life-threatening complication of neutropenia, frequently requires hospital admission, while fatigue and weakness from anaemia reduce patient's capacity for activity. Pegfilgrastim and darbepoetin alfa, were designed to simplify and optimise treatment for patients with cancer. Once-per-cycle pegfilgrastim is as effective as daily filgrastim with respect to duration of severe neutropenia (DSN) and may have a lower incidence of febrile neutropenia than filgrastim. Darbepoetin alfa has enhanced biological activity and a serum terminal half-life three-fold longer than that of erythropoietin (EPO), which translates into rapid and sustained correction of anaemia with less frequent dosing. These novel cytokines have the potential to simplify the management of neutropenia and anaemia with fewer injections and less disruption to patients daily lives. PMID- 14563521 TI - Systemic chemotherapy in elderly patients with locally advanced and/or inoperable squamous cell carcinoma of the head and neck: impact of anemia and role of recombinant human erythropoietin. AB - A review of the incidence and management of anemia in elderly patients with head and neck carcinoma treated with systemic chemotherapy. The role of recombinant human erythropoietin in preventing or correcting chemotherapy-related anemia has been focused. Data concerning the prospective use of recombinant human erythropoietin (rhEpo) in a series of unfit elderly patients (EPs) treated with carboplatin plus 5-fluorouracil. Patients were randomly assigned to receive subcutaneous rhEpo 10,000U three times per week (TIW) (23 elderly patients) or no treatment (22 control patients). Recombinant hEpo was able to prevent anemia and to reduce transfusional requirements in treated patients as compared to untreated controls with a statistically significant difference. rhEpo also caused a positive effect on quality of life (QoL) parameters. PMID- 14563522 TI - Fatigue and aging. AB - Fatigue is the most common chronic symptom of cancer and the symptom most likely to disrupt the patient's activity and to cause disability.Fatigue affects older cancer patients to the same order of magnitude as younger ones and appears to be related to different factors such as anemia, depression, and reduced neuromuscular energy production. Interestingly, some studies have suggested that the duration of fatigue may be age-related. In the elderly, fatigue may cause functional dependence; functional dependence may lead to interruption of treatment, decline in quality of life and expensive home care. Available interventions include reduction of centrally acting drugs, management of anemia and metabolic abnormalities, and management of depression, treatment of sleep disorders, correction of endocrine abnormalities and realistic exercise programmes. PMID- 14563523 TI - Supportive care of the older cancer patient. AB - Aging is associated with decreased functional reserve of multiple organ systems and with changes in the pharmacokinetics and pharmacodinamics of drugs. Older individuals express enhanced susceptibility to the complications of cytotoxic chemotherapy, especially to myleotoxicity, mucositis, cardiotoxicity and neurotoxicity. The management of older individuals with chemotherapy involves then prevention of these complications. General precautions include proper patient selection, based on the comprehensive geriatric assessment (CGA), dose adjustment for agents that are renally excreted to the patient creatinine clearance and maintenance of hemoglobin levels > or =12 g/dl. Filgrastim and pegfilgrastim proved effective in reducing by 50-75% the risk of neutropenic fever in older individuals treated with CHOP and CHOP-like chemotherapy and should be used for the prophilaxis of infections. When feasible, the oral agent capecitabine, should be used in lieu of intravenous fluorinated pyrimidines, to prevent mucositis. In patients at risk of cardiomyopathy from anthracyclines, dexrazoxane or liposomal compounds may be indicated. When toxicity is properly prevented, cytotoxic chemotherapy may be as effective in older individuals as it is in the younger ones. PMID- 14563524 TI - Cost analyses of granulocyte colony stimulating factor: a focus on older patients with cancer. AB - Granulocyte colony stimulating factor (G-CSF) can reduce the duration of neutropenia following intensive chemotherapy in a variety of settings. The majority of these studies have focused on small cell lung cancer. We review herein published information on costs and cost-effectiveness of G-CSF from clinical studies with patients with small cell lung cancer (SCLC). Medline and Healthstar databases were searched for original research articles that contain cost or cost-effectiveness analyses on the subject. The cost of adjunct treatment with G-CSF was evaluated from four studies of SCLC patients. Benefits included decreased rates of febrile neutropenic events, while cost savings were not identified in three of four studies. These data may be useful to physicians faced with concerns over clinical and economic factors associated with G-CSF use as adjunct therapy for SCLC. PMID- 14563525 TI - Translocation analysis by the FISH-painting method for retrospective dose reconstruction in individuals exposed to ionizing radiation 10 years after exposure. AB - Fluorescence in situ hybridization (FISH) is a powerful method largely used for detecting chromosomal rearrangements, translocations in particular, which are important biomarkers for dose assessment in case of human exposure to ionizing radiation. To test the possibility of using the translocation analysis by FISH painting method in retrospective dose assessment, we carried out in vitro experiments in irradiated human lymphocytes, in parallel with the analysis of translocations in lymphocytes from 10 individuals, who were exposed to 137cesium in the Goiania (Brazil) accident (samples collected 10 years after exposure). The in vitro dose-response curve for the genomic translocation frequencies (FGs) fits a linear quadratic model, according to the equation: Y=0.0243X(2)+0.0556X. The FG values were also calculated for the individuals exposed to 137cesium, ranging from 0.58 to 5.91 per 100 cells, and the doses were estimated and compared with the results obtained by dicentric analysis soon after the accident, taking the opportunity to test the validity of translocation analysis in retrospective biodosimetry. A tentative of retrospective dosimetry was performed, indicating that the method is feasible only for low level exposure (below 0.5Gy), while for higher doses there is a need to apply appropriate correction factors, which take into consideration mainly the persistence of chromosomal translocations along with time, and the influence of endogenous and exogenous factors determining the inter-individual variability in the cellular responses to radiation. PMID- 14563526 TI - Prevalence of aberrant methylation of p14ARF over p16INK4a in some human primary tumors. AB - The INK4a/ARF locus encodes two unrelated tumor suppressor proteins, p16INK4a and p14ARF, which participate in the two main cell-cycle control pathways, p16-Rb and p14-p53. Methylation of CpG promoter islands has been described as a mechanism of gene silencing. Exon 1 of the p16INK4a gene and the p14ARF promoter gene reside within CpG islands. Therefore, both can become methylated de novo and silenced. It has recently been proposed that the methylation changes in certain genes could be used as molecular markers for the detection of almost all forms of human cancer. Here, we analyzed concomitantly in each tumor sample and normal tissue the methylation status of p16INK4a and p14ARF by methylation-specific PCR (MSP) in 100 breast, 95 colon and 27 bladder carcinomas. A series of clinicopathological parameter were obtained from the medical records of the patients, p14ARF showed a higher rate of hypermethylation than p16INK4a in all three tumor types. p16INK4a and p14ARF aberrant methylation was significantly correlated with poor prognosis clinicopathological parameters of the three tumor types. We conclude that both p16INKa and p14ARF hypermethylation may be involved in breast, colon and bladder carcinogenesis, with special emphasis on the role of the lesser studied p14ARF gene, and that tumors with aberrant methylation in the two genes were associated with worse prognosis. PMID- 14563527 TI - Photomutagenicity of cosmetic ingredient chemicals azulene and guaiazulene. AB - The photomutagenicity of the popular skin conditioning agents azulene and guaiazulene were tested in Salmonella typhimurium TA98, TA100 and TA102. Following irradiation with UVA and/or visible light, both azulene and guaiazulene exhibited mutagenicity 4-5-fold higher than the spontaneous background mutation. In contrary, naphthalene, a structural isomer of azulene, was not photomutagenic under the same conditions. Azulene was photomutagenic when irradiated with UVA light alone, visible light alone, or a combination of UVA and visible light. Azulene and guaiazulene are not mutagenic when the experiment is conducted with the exclusion of light. Therefore, extreme care must be taken when using cosmetic products with azulene/guaiazulene as ingredients since after applying these products on the skin, exposure to sunlight is inevitable. PMID- 14563528 TI - Linoleic acid and antioxidants protect against DNA damage and apoptosis induced by palmitic acid. AB - Polyunsaturated fats are the main target for lipid peroxidation and subsequent formation of mutagenic metabolites, but diets high in saturated fats are more strongly associated with adverse health effects. We show that the common saturated fatty acid, palmitic acid, is a potent inducer of DNA damage in an insulin-secreting cell line, and in primary human fibroblasts. Damage is not associated with upregulation of inducible nitric oxide synthase, but is prevented by two different antioxidants, alpha-lipoic acid and 3,3'-methoxysalenMn(III) (EUK134), which also partly prevent palmitic acid-induced apoptosis and growth inhibition. Since mutagenic metabolites can be formed from peroxidation of polyunsaturated fatty acids, co-administration of palmitic and a polyunsaturated fatty acid might be particularly harmful. Palmitic acid-induced DNA damage is instead prevented by linoleic acid, which is acting here as a protective agent against oxidative stress, rather than as a source of mutagenic metabolites. These results illustrate the complexity of the relationship of dietary fat intake to genotoxicity. PMID- 14563529 TI - DNA strand breaks induced by the anti-topoisomerase II bis-dioxopiperazine ICRF 193. AB - The bis-dioxopiperazine ICRF-193 has long time been considered as a pure topoisomerase II catalytic inhibitor able to exert its inhibitory effect on the enzyme without stabilization of the so-called cleavable complex formed by DNA covalently bound to topoisomerase II. In recent years, however, this concept has been challenged, as a number of reports have shown that ICRF-193 really "poisons" the enzyme, most likely through a different mechanism from that shown by the classical topoisomerase II poisons used in cancer chemotherapy. In the present investigation, we have carried out a study of the capacity of ICRF-193 to induce DNA strand breaks, as classical poisons do, in cultured V79 and irs-2 Chinese hamster lung fibroblasts using the comet assay and pulsed-field gel electrophoresis (PFGE). Our results clearly show that ICRF-193 readily induces breakage in DNA through a mechanism as yet poorly understood. PMID- 14563530 TI - Alleviation of mutagenic effects of polycyclic aromatic agents (quinacrine mustard, ICR-191 and ICR-170) by caffeine and pentoxifylline. AB - Previous studies performed by others indicated that apart from its other biological effects, caffeine (CAF) may have a role in protection of organisms against cancer. However, biological mechanism of this phenomenon remained unknown. Recent studies suggested that caffeine can form stacking (pi-pi) complexes with polycyclic aromatic chemicals. Therefore, one might speculate that effective concentrations of polycyclic aromatic mutagens could be reduced in the presence of caffeine. Here we demonstrate that caffeine and another xanthine, pentoxifylline (PTX), effectively alleviate mutagenic action of polycyclic aromatic agents (exemplified by quinacrine mustard (QM), 2-methoxy-6-chloro-9-(3 (2-chloroethyl)aminopropylamino)acridine.2HCl (ICR-191) and 1,3,7-propanediamine N-(2-chloroethyl)-N'-(6-chloro-2-methoxy-9-acridinyl)-N-ethyl.2HCl (ICR-170)), but not of aliphatic mutagens (exemplified by mechlorethamine), in the recently developed mutagenicity test based on bacterium Vibrio harveyi. Biophysical studies indicated that caffeine and pentoxifylline can form stacking complexes with the aromatic agents mentioned above. Molecular modeling also confirmed a possibility of stacking interactions between examined molecules. PMID- 14563531 TI - Induction of structural and numerical changes of chromosome, centrosome abnormality, multipolar spindles and multipolar division in cultured Chinese hamster V79 cells by exposure to a trivalent dimethylarsenic compound. AB - Dimethylarsine iodide (DMI) was used as a model compound of trivalent dimethylarsenicals [DMA(III)], and the biological effects were extensively investigated in cultured Chinese hamster V79 cells. When the cytotoxic effects of DMA(III) were compared with those of inorganic arsenite and dimethylarsinic acid [DMA(V)], DMA(III) was about 10,000 times more potent than DMA(V), and it was even 10 times more toxic than arsenite. Depletion of cell glutathione (GSH) did not influence the cytotoxic effects of DMA(III), whereas it enhanced the cytotoxicity of arsenite. Chromosome structural aberrations, such as gaps, breaks and pulverizations, and numerical changes, such as aneuploidy, hyper- and hypo tetraploidy, were induced by DMA(III) in a concentration-dependent manner. Mitotic index increased 9-12h after the addition of DMA(III), and then declined. By contrast, the incidence of multinucleated cells increased conversely with the decrease in mitotic index at and after 24h of exposure. The mitotic cell-specific abnormality of centrosome integrity and multipolar spindles were induced by DMA(III) in a time- and concentration-dependent manner. Moreover, DMA(III) caused abnormal cytokinesis (multipolar division) at concentrations that were effective in causing centrosome abnormality, multipolar spindles and aneuploidy. These results showed that DMA(III) was genotoxic on cultured mammalian cells. Results also suggest that DMA(III)-induced multipolar spindles and multipolar division may be associated with the induction of aneuploidy. In addition, the centrosome may be a primary target for cell death via multinucleated cells. PMID- 14563532 TI - The major surface protease (MSP or GP63) of Leishmania sp. Biosynthesis, regulation of expression, and function. AB - Leishmania sp. are digenetic protozoa that cause an estimated 1.5-2 million new cases of leishmaniasis per year worldwide. Among the molecular factors that contribute to Leishmania sp. virulence and pathogenesis is the major surface protease, alternately called MSP, GP63, leishmanolysin, EC3.4.24.36, and PSP, which is the most abundant surface protein of leishmania promastigotes. Recent studies using gene knockout, antisense RNA and overexpression mutants have demonstrated a role for MSP in resistance of promastigotes to complement-mediated lysis and either a direct or indirect role in receptor-mediated uptake of leishmania. The MSP gene clusters in different Leishmania sp. include multiple distinct MSPs that tend to fall into three classes, which can be distinguished by their sequences and by their differential expression in parasite life stages. Regulated expression of MSP class gene products during the parasite life cycle occurs at several levels involving both mRNA and protein metabolism. In this review we summarize advances in MSP research over the past decade, including organization of the gene families, crystal structure of the protein, regulation of mRNA and protein expression, biosynthesis and possible functions. The MSPs exquisitely demonstrate the multiple levels of post-transcriptional gene regulation that occur in Leishmania sp. and other trypanosomatid protozoa. PMID- 14563533 TI - Maurer's clefts--a novel secretory organelle? AB - During intra-erythrocytic development, the human malarial parasite Plasmodium falciparum extensively remodels its adopted cellular home by exporting proteins beyond the confines of its own plasma membrane, but is, however, faced with a major problem: the lack of an endogenous protein trafficking machinery within the host erythrocyte. Thus, in order to export proteins the parasite has to install its own protein export system within the host erythrocyte. A growing body of evidence suggests that Maurer's clefts, parasite-derived membranous structures in the cytosol of the host cell, are a crucial component of this protein sorting and trafficking machinery. In this review we summarize our current understanding of the ultra-structure of Maurer's clefts and their role in protein transport process. PMID- 14563534 TI - Extracellular lysines on the plasmodial surface anion channel involved in Na+ exclusion. AB - The human malaria parasite, Plasmodium falciparum, induces an unusual ion channel, the plasmodial surface anion channel (PSAC), on its host red blood cell (RBC) membrane. PSAC has a broad selectivity with permeability to anions, sugars, amino acids, purines, and certain vitamins, suggesting a role in nutrient acquisition by the intracellular parasite. Permeating solutes cover a range of molecular sizes and may be either neutral or carry a net negative or positive charge. Despite this broad selectivity, PSAC must efficiently exclude Na+ to avoid osmotic lysis of infected RBCs in the bloodstream. Here, we used amine reactive N-hydroxysulfosuccinimide esters to probe PSAC's unusual selectivity. PSAC permeation rates, measured with both a kinetic osmotic lysis assay and single-channel patch-clamp, irreversibly decrease after treatment with these reagents. Sequential labelings with different esters and the effects of their chain length suggest that PSAC has multiple lysine residues near its extracellular pore mouth and that inhibition occurs via steric hindrance of its pore by the amide-linked side chain. When combined with the effects of pH on permeation, these findings implicate a combination of cation repulsion by pore mouth charges and a weak binding site for permeant solutes in PSAC's broad selectivity yet effective exclusion of Na+. PMID- 14563535 TI - A biotin derivative blocks parasite induced novel permeation pathways in Plasmodium falciparum-infected erythrocytes. AB - The malaria parasite Plasmodium falciparum infects human erythrocytes, and it induces an increased rate of uptake into the infected cell of a range of solutes, including essential nutrients required for parasite development. Several models have been proposed for the mechanism(s) underlying parasite-induced solute uptake, each differing with respect to the site of entry into infected cells. We show that a biotin derivative that is excluded from non-infected erythrocytes gains access to infected erythrocytes via a pathway that is inhibited by compounds shown previously to block the pathways responsible for the increased uptake of solutes. The derivative was found to bind erythrocyte cytoskeletal proteins and to hemoglobin, providing evidence that the novel pathways are in the erythrocyte membrane and allow direct access of solutes to the erythrocyte cytosol. The derivative inhibited its own uptake and blocked the parasite-induced transport of other solutes. In whole-cell patch-clamp analyses, biotinylation of infected erythrocytes caused significant decrease in a parasite-induced outward rectifying conductance. In vitro, biotinylation of trophozoite-stage parasitized erythrocytes delayed parasite development. Treatment of infected cells in the final developmental stage abrogated the parasite's ability to complete development. The data are consistent with the novel pathways playing an important role in parasite growth. PMID- 14563536 TI - Cloning and characterisation of the UDP-glucose 4'-epimerase of Trypanosoma cruzi. AB - Trypanosoma cruzi incorporates galactose into many of its cell-surface glycoconjugates but it is unable to transport this sugar through its hexose transporter. Epimerisation of UDP-glucose to UDP-galactose by UDP-glucose 4' epimerase may be the only way that the parasites can obtain galactose. Here, we describe cloning the T. cruzi UDP-Glc 4'-epimerase (TcGALE) gene and show that it is functional by complementing an Escherichia coli epimerase-deficient strain. The T. cruzi GALE gene encodes a 42.4 kDa protein and the recombinant protein expressed in E. coli is a homodimer in solution with a specific activity of 3.8 U mg(-1) and K(m) for UDP-Gal of 114 microM. Unlike the human epimerase, T. cruzi UDP-Glc 4'-epimerase is unable to inter-convert UDP-N-acetylglucosamine and UDP-N acetylgalactosamine. This may explain why T. cruzi initiates O-glycosylation of its abundant GPI-anchored surface mucins via GlcNAcalpha1-O-Thr/Ser rather than the GalNAcalpha1-O-Thr/Ser linkage that is common for mucins from many other eukaryotes. PMID- 14563537 TI - Intraerythrocytic Plasmodium falciparum incorporates extraneous fatty acids to its lipids without any structural modification. PMID- 14563538 TI - Developmental exposure to cadmium alters responsiveness to cocaine in the rat. AB - The purpose of the present report was to investigate the potential interactive relation between perinatal (gestation/lactation) cadmium exposure and changes in responsiveness to cocaine. In Experiment 1, adult female rats were exposed to a diet containing 50 ppm cadmium (as cadmium chloride) or a diet containing no added cadmium for 30 days prior to breeding with nonexposed males. The metal exposure regimen continued throughout gestation, and for 15 days of lactation, at which time all animals were placed on standard rat chow diets containing no added cadmium for the remainder of the investigation. Atomic absorption assays confirmed that cadmium concentrations were significantly elevated in metal exposed dams, littermates, and test animals. Offspring were weaned on postnatal day (PND) 21 and commenced cocaine sensitization testing on PND 70. Testing operations for controls and animals perinatally exposed to cadmium consisted of 21 daily i.p. injections of vehicle (saline) or 10 mg/kg cocaine HCl, and subsequent recording of locomotor activity. Subsequently, across successive days, all animals received 0, 10, and 20 mg/kg cocaine challenges. The results showed that cocaine sensitization was attenuated in animals perinatally exposed to cadmium. A similar pattern of antagonism was observed in Experiment 2 where a higher dose of cocaine was required to produce conditioned place preference (CPP) in cadmium-exposed animals. The implications of these findings with respect to the interactive role of cadmium in the dynamics of cocaine use/abuse remain unclear. PMID- 14563539 TI - Evaluation of buprenorphine maintenance treatment in a French cohort of HIV infected injecting drug users. AB - BACKGROUND: Buprenorphine was approved in France for treating opiate dependence in July 1995 and can be prescribed by general practitioners (GPs). Most studies assessing buprenorphine maintenance treatment (BMT) outcomes have taken place in GP settings. An evaluation of BMT outcomes in patients already followed for their HIV-infection could supply additional information about the changes in addictive practices in a non-GP setting. METHODS: We assessed BMT discontinuations and the course of self-reported addictive behaviours and characteristics associated with buprenorphine-injection misuse in 114 HIV-infected patients on BMT who were followed in a hospital-based outpatient department. RESULTS: The continuous series of follow-up visits at which these 114 patients reported regular buprenorphine prescriptions accounted for 237.5 person-years of observation, i.e. 475 follow-up visits. Of the 114 patients on BMT, 43% continued BMT throughout the follow-up, 40% stopped it, and results for 17% were not available either because they did not answer the self-administered questionnaire (5%) or because they were lost to follow-up (12%). Addictive behaviours declined but buprenorphine injection misuse remained stable. Depression measured by the CESD score (RR=1.04 95%CI [1.01-1.06]), cocaine use (RR=2.48 95%CI [1.31-4.68]) and alcohol consumption exceeding 4 alcohol units (AU) per day (RR=2.29, 95%CI [1.17 4.46]) were independently associated with buprenorphine injection misuse among stabilised BMT patients. CONCLUSIONS: Despite the reduction in drug injection after starting BMT, buprenorphine injection misuse mainly involves patients with characteristics of severe addiction. Better monitoring of the illicit drug use patterns of patients on BMT may suggest new medical strategies for GPs to improve BMT outcomes. PMID- 14563540 TI - Distribution of deaths by unintentional illicit drug overdose in Italy based on periodicity over time, 1984-2000. AB - AIM: To assess whether substance-abusing people have a higher risk of dying from unintentional acute intoxication in specific periods of the year. METHODS: A total of 15,792 deaths (males=14,259; females=1533) were ascertained to be due to unintentional acute intoxication by illicit drugs in Italy in the study period (1984-2000). Distribution of deaths by month was studied with circular statistics techniques, based on the maximization of mean vector length (MMVL) method. The Rayleigh test was used to verify the hypothesis of uniform distribution of the events across some relevant periods. Density estimates for significant periodicities were derived with the kernel method for circular variables. RESULTS: Over the study period there was a steady increase in the number of deaths by overdose in Italy, affecting both genders, but more evident among males. The monthly distribution of deaths over the study period followed an uneven trend, with a slightly higher risk in the period between December and January, and in August, in both genders. The monthly distribution of deaths due to unintentional acute intoxication by illicit drugs appears influenced by non casual oscillations following a thrice-yearly cycle, interlaced with a significantly more evident 6-monthly recurrence. CONCLUSIONS: There does seem to be a higher risk of dying from unintentional overdose in specific periods of the year. Better knowledge of the factors affecting the risk of mortality among substance-abusing people--due to changes in environmental or biological rhythms- could allow prediction of negative events, hence their prevention. PMID- 14563541 TI - Reinforcing, subjective, and physiological effects of MDMA in humans: a comparison with d-amphetamine and mCPP. AB - 3,4-methylenedioxymethamphetamine (MDMA) is a widely used drug of abuse chemically related to both the amphetamines and mescaline. Laboratory animal studies have shown that MDMA is a potent re-uptake inhibitor and releaser of dopamine and serotonin. Although the subjective and physiological effects of MDMA have been compared to d-amphetamine in humans, no direct comparison with a serotonin releasing agent has been reported and reinforcing effects have not been evaluated. In this paper we report a direct comparison of the reinforcing, subjective, and physiological effects of MDMA (1 and 2 mg/kg) to d-amphetamine (10 and 20 mg), to metachlorophenylpiperazine (mCPP--a serotonin releasing agent (0.5 and 0.75 mg/kg)), and to placebo using a within-subject design in 12 volunteers with moderate MDMA experience. Both the high dose of d-amphetamine and MDMA showed significant reinforcing effects as indicated by high cross-over values on the multiple choice procedure compared to all other treatments. All three drugs showed dose-dependent changes in subjective effects whereas physiological effects were most pronounced for MDMA with almost no changes seen with mCPP. The subjective effects of MDMA were similar both to those of mCPP and d-amphetamine, suggesting that both dopamine and serotonin systems are involved in mediating these effects. In contrast, only the dopaminergic agents, d amphetamine and MDMA, had reinforcing effects. PMID- 14563542 TI - An enumeration method of determining the prevalence of users and operatives of cocaine and heroin in Central Harlem. AB - INTRODUCTION: The investigators estimated the number and characteristics of crack, powder cocaine and heroin users and distributors ("operatives") in Central Harlem. METHODS: Central Harlem was divided into 45 primary sampling units (PSUs) with 2 years of drug allegation data from the New York Police Department (NYPD) organized in three strata, those having high, middle or low levels of allegations. In nine randomly selected PSUs, interviewers employed chain referral sampling procedures that were steered by using a nomination technique. RESULTS: Within sampled PSUs, 657 respondents nominated 5756 others for a total of 6413. Seven indicators identified 1007 unique individuals among 2835 nominated by two or more respondents, totaling to 4585 unique persons. These drug users and operatives were divided by the total number of persons in these PSUs according to the 1990 census to estimate the number of drug users and distributors in the 36 PSUs not sampled and for all 98,725 residents of Central Harlem. Approximately 13.4% of Central Harlem residents were estimated to be users of crack, powder cocaine and/or heroin with no roles in drug distribution. Approximately 6.7% of Central Harlem residents were estimated to be operatives of these drugs, some of whom may also have been users. CONCLUSIONS: Enumeration techniques can be a basis for projecting prevalence estimates for crack, powder cocaine and heroin users and operatives in a large urban geographic area. PMID- 14563543 TI - A randomised controlled trial of methadone maintenance treatment versus wait list control in an Australian prison system. AB - OBJECTIVES: The aim was to determine whether methadone maintenance treatment reduced heroin use, syringe sharing and HIV or hepatitis C incidence among prisoners. METHODS: All eligible prisoners seeking drug treatment were randomised to methadone or a waitlist control group from 1997 to 1998 and followed up after 4 months. Heroin use was measured by hair analysis and self report; drugs used and injected and syringe sharing were measured by self report. Hepatitis C and HIV incidence was measured by serology. RESULTS: Of 593 eligible prisoners, 382 (64%) were randomised to MMT (n=191) or control (n=191). 129 treated and 124 control subjects were followed up at 5 months. Heroin use was significantly lower among treated than control subjects at follow up. Treated subjects reported lower levels of drug injection and syringe sharing at follow up. There was no difference in HIV or hepatitis C incidence. CONCLUSIONS: Consideration should be given to the introduction of prison methadone programs particular where community based programs exist. PMID- 14563544 TI - Cause and manner of death in drug-related fatality: an analysis of drug-related deaths recorded by coroners in England and Wales in 2000. AB - This study investigated causes and manner of drug-related fatalities recorded in 2000 in the United Kingdom, measuring the 'masked' manner of death in cases typically recorded as overdose. A retrospective cohort study was used of 1037 cases of accidental drug-related fatalities reported by coroners in England and Wales to the National Programme of Substance Abuse Deaths. Whilst 802 cases were identified as direct acute overdose, representing 77% of the total accidental deaths, 23% of 'overdose' fatalities were caused by asphyxiation (7%), drug related medical conditions (7%), non-drug-related conditions (4%), traumatic accidents (3%) and infections (2%). Younger people show higher risk of overdose and asphyxiation; older people show higher risk from pre-existing medical conditions. This study not only confirmed the high risk of overdose associated with heroin and polydrug use, but it also identified other high fatality risk factors for heroin/morphine users such as contracting an acute infection leading to septicaemia or endocarditis, or contracting a chronic infection such as HIV, HBV or HCV. In contrast, stimulants particularly featured in traumatic accidents, with amphetamine use most associated with cardio-vascular fatality. These findings highlight the 'masked' manner of death in cases commonly recorded as overdose and demonstrate the need for a more-detailed and systematic method of recording drug-related deaths in order to inform drug education and harm reduction strategies. PMID- 14563545 TI - Pharmacokinetics of high-dose buprenorphine following single administration of sublingual tablet formulations in opioid naive healthy male volunteers under a naltrexone block. AB - Sublingual buprenorphine formulations have been developed as treatments for opioid dependence. In three studies, opioid naive healthy male subjects received Subutex tablets (buprenorphine 2 and 8 mg [N=27] or 12 and 16 mg [N=27]) or Suboxone (two formulations) tablets (buprenorphine 8 mg/naloxone 2 mg [N=36]) sublingually, under a naltrexone block for assessment of buprenorphine pharmacokinetics and tablet disintegration times. Plasma buprenorphine was quantified up to 72 h post-dose using a sensitive LC-MS/MS assay. Mean Cmax values ranged from 1.6 to 6.4 ng/ml and tmax from 0.5 to 3 h. Concentrations declined bi-exponentially and fluctuations after a meal suggested enterohepatic recirculation of buprenorphine. The terminal half-life was approximately 26 h (range 9-69). Cmax and AUC appeared to increase in proportion to Subutex dose over 8-16 mg. The Suboxone formulations were bioequivalent. The least squares mean (90% CI) treatment ratio for Cmax was 1.00 (0.92-1.10) and AUC was 1.00 (0.95-1.06). Median times of disintegration were similar for all doses and formulations (range 6-12 min). Sublingual buprenorphine, up to 40 times the 400 microg analgesic dose, was well tolerated in these opioid naive subjects, as administration of naltrexone 50-150 mg was sufficient to attenuate anticipated adverse effects in this population of subjects. PMID- 14563546 TI - Comparative pharmacodynamics and pharmacokinetics of methadone and slow-release oral morphine for maintenance treatment of opioid dependence. AB - BACKGROUND: Slow-release oral morphine (SROM) has been proposed as an alternative maintenance pharmacotherapy to methadone for treatment of opioid dependence. However, the pharmacodynamics and pharmacokinetics of SROM have not been previously assessed in a methadone maintenance population. METHODS: In 14 methadone maintenance patients reporting adequate (holders, n=7) or inadequate (non-holders, n=7) withdrawal suppression between doses, plasma drug concentrations and indices of opioid effect (withdrawal severity, pupil diameter, and respiratory rate) were determined across a 24 h inter-dosing interval on one occasion at steady-state for methadone and SROM (once-daily Kapanol) using an open-label, crossover design. RESULTS: Opioid effects were of a similar overall magnitude following dosing for each drug and showed an inverse association with plasma drug concentrations, which peaked later for morphine compared to (R) methadone (6.5 +/- 2.3 h vs. 2.5 +/- 1.4 h, P<0.001). For methadone non-holders, the number of self-reported opioid withdrawal symptoms prior to dosing was less for SROM compared to methadone (3.4 +/- 2.6 vs. 9.0 +/- 3.2, P<0.01); for holders it was approximately equal (4.4 +/- 3.0 vs. 4.0 +/- 3.7, P<0.76). Maintenance doses were more strongly related to plasma drug concentrations for SROM compared to methadone and may thus be more predictive of therapeutic response for the former medication. Twelve of the 14 subjects preferred SROM to methadone (P=0.01). CONCLUSIONS: The pharmacodynamics and pharmacokinetics of SROM support its use as an alternative once-daily agonist option in the treatment of opioid dependence, particularly for patients responding poorly to methadone maintenance treatment. PMID- 14563547 TI - Role of MADS box proteins and their cofactors in combinatorial control of gene expression and cell development. AB - In all organisms, correct development, growth and function depends on the precise and integrated control of the expression of their genes. Often, gene regulation depends upon the cooperative binding of proteins to DNA and upon protein-protein interactions. Eukaryotes have widely exploited combinatorial strategies to create gene regulatory networks. MADS box proteins constitute the perfect example of cellular coordinators. These proteins belong to a large family of transcription factors present in most eukaryotic organisms and are involved in diverse and important biological functions. MADS box proteins are combinatorial transcription factors in that they often derive their regulatory specificity from other DNA binding or accessory factors. This review is aimed at analyzing how MADS box proteins combine with a variety of cofactors to achieve functional diversity. PMID- 14563548 TI - Fibroblast growth factor signaling controlling osteoblast differentiation. AB - Fibroblast growth factors (FGFs) play important roles in skeletal development and postnatal osteogenesis. FGF signaling controls bone formation by regulating the expression of various genes involved in osteoprogenitor cell replication, osteoblast differentiation and apoptosis. Recent genetic manipulation of FGF expression in mice and studies of the phenotype induced by gain-of-function mutations in FGF receptors in humans revealed the important role of FGF signaling in osteoblast function and differentiation. Additionally, cell biology studies allowed to identify some signaling pathways that are involved in the control of FGF actions in osteoblasts. This led to a better understanding of the functional role of FGF signaling in the control of gene expression in osteoblasts. The elucidation of molecular mechanisms by which FGF signaling promotes osteoblast gene expression and differentiation may help to find novel molecular targets and develop new therapeutic approaches to promote bone formation in human bone disorders. PMID- 14563549 TI - DNA transfer from chloroplast to nucleus is much rarer in Chlamydomonas than in tobacco. AB - By transforming chloroplasts with an antibiotic-resistance gene under the control of a nuclear-specific promoter, we employed a selection scheme to detect the transfer of DNA from the chloroplast to the nucleus in the green alga Chlamydomonas reinhardtii. Among several billion homoplasmic cells tested, we were unable to detect any stable nuclear integration of chloroplast DNA under normal growth conditions or under stress conditions. This contrasts with results reported for the transfer of DNA from chloroplast to nucleus in higher plants and from mitochondrion to nucleus in Saccharomyces cerevisiae. Furthermore, we were unable to detect chloroplast DNA-derived sequences among nuclear genome data for C. reinhardtii, which also contrasts with the situation in higher plants. Taken together, these findings suggest that there is presently little, if any, movement of DNA from chloroplast to nucleus in C. reinhardtii, which may reflect the ultrastructure of the C. reinhardtii cell. PMID- 14563550 TI - Characterization of chicken CENP-A and comparative sequence analysis of vertebrate centromere-specific histone H3-like proteins. AB - Centromere protein A (CENP-A) is a centromere-specific histone H3 variant conserved amongst all eukaryotes. We have isolated the chicken gene for CENP-A (GgCENP-A). It encodes a 131-amino-acid polypeptide that possesses an average identity of 54% with human CENP-A, reaching 69% in the histone-fold domain. The gene spans 1.7 kb of genomic DNA and contains four exons that range in size from 78 to 186 bp. The exon/intron organisation of the chicken gene is conserved with its mammalian counterparts in the carboxy-terminal histone-fold domain (exons 2 to 4), consistent with the strong conservation of this domain at the amino acid level. Sequence analysis of the chicken CENP-A locus revealed that the gene is located within the class III genes of the major histocompatibility complex (MHC), and extended the previously defined limit of the compact chicken MHC complex. We compared the sequences of CENP-A from mammals, chicken and fishes and thereby identified conserved motifs in the otherwise variable amino-terminal tail that may be important for functional reasons. We also identified evolutionarily variable regions within the conserved histone-fold domain. We found that loop 1 between the first and second alpha-helix is the region that diverged most widely. This finding is in agreement with evolutionary studies in Drosophila species, and suggests that this domain could play a role in species-specific centromere targeting of CENP-A. In addition, protein sequence comparison of several vertebrate species revealed that the RT-PCR strategy we have developed for isolating the chicken centromeric histone H3 variant gene should be applicable to the isolation of CENP-A from a wide range of vertebrates. PMID- 14563551 TI - The human SUMF1 gene, required for posttranslational sulfatase modification, defines a new gene family which is conserved from pro- to eukaryotes. AB - Recently, the human C(alpha)-formylglycine (FGly)-generating enzyme (FGE), whose deficiency causes the autosomal-recessively transmitted lysosomal storage disease multiple sulfatase deficiency (MSD), has been identified. In sulfatases, FGE posttranslationally converts a cysteine residue to FGly, which is part of the catalytic site and is essential for sulfatase activity. FGE is encoded by the sulfatase modifying factor 1 (SUMF1) gene, which defines a new gene family comprising orthologs from prokaryotes to higher eukaryotes. The genomes of E. coli, S. cerevisiae and C. elegans lack SUMF1, indicating a phylogenetic gap and the existence of an alternative FGly-generating system. The genomes of vertebrates including mouse, man and pufferfish contain a sulfatase modifying factor 2 (SUMF2) gene encoding an FGE paralog of unknown function. SUMF2 evolved from a single exon SUMF1 gene as found in diptera prior to divergent intron acquisition. In several prokaryotic genomes, the SUMF1 gene is cotranscribed with genes encoding sulfatases which require FGly modification. The FGE protein contains a single domain that is made up of three highly conserved subdomains spaced by nonconserved sequences of variable lengths. The similarity among the eukaryotic FGE orthologs varies between 72% and 100% for the three subdomains and is highest for the C-terminal subdomain, which is a hotspot for mutations in MSD patients. PMID- 14563552 TI - Induction of endogenous telomerase (hTERT) by c-Myc in WI-38 fibroblasts transformed with specific genetic elements. AB - Elucidation of the mechanisms governing expression of the human telomerase reverse transcriptase (hTERT) is important for understanding cancer pathogenesis. Approximately 90% of tumors express hTERT, the major catalytic component of telomerase. Activation of telomerase is an early event, and high levels of this activity correlate with poor prognosis. Recent studies have shown that the transcription factors c-Myc and Mad1 activate and repress hTERT, respectively. It is not clear how these transcription factors compete for the same recognition sequence in the hTERT core promoter region. Studies have shown that the combined expression of SV40 large T antigen (T-Ag), hTERT, and H-Ras is able to transform human cells. In this study, we used a distinct human cell type, WI-38 fetal lung fibroblasts used extensively for senescence studies. We transduced cells with amphotropic retroviral constructs containing SV40 T antigen, hTERT, and activated H-ras. Transduced cells exhibited anchorage independence in soft agar and expressed increased levels of c-Myc and endogenous hTERT. These effects were observed by 25 population doublings (PDs) following the establishment of the neoplastic cell line. During the process of transformation, we observed a switch from Mad1/Max to c-Myc/Max binding to oligonucleotide sequences containing the hTERT promoter distal and proximal E-boxes. c-Myc can bind specifically to the hTERT promoter in vitro, indicating that c-Myc expression in tumors may account for the increased expression of hTERT observed in vivo. These findings indicate that the widely used model system of WI-38 fibroblasts can be employed for transformation studies using defined genetic elements and that the endogenous hTERT and c-Myc are induced in these cells during early tumorigenesis. Such studies should have important implications in the mechanisms of hTERT and c-Myc induction in the beginning stages of tumorigenesis and facilitate extension of these studies to novel models of tumorigenesis in cellular senescence. PMID- 14563553 TI - Cloning of CnLYSO1, a novel extracellular lysophospholipase of the pathogenic fungus Cryptococcus neoformans. AB - We cloned a novel lysophospholipase (CnLYSO1) from Cryptococcus neoformans var. grubii by PCR amplification and a cDNA library screen. The open reading frame (ORF) of 1278 nucleotides coded for a predicted 426-amino-acid protein (CnLyso1p) with two highly conserved GXSXG lipase-specific catalytic motifs and a molecular weight of 48.3 kDa. CnLyso1p exhibited 14% and 21% identity to Arabidopsis thaliana and human lysophospholipases, respectively. Immunoprecipitation and Western blot analyses indicated that CnLyso1p was secreted as a high molecular weight protein of 97-140 kDa. CnLYSO1 expressed in a phospholipase B-null mutant of Saccharomyces cerevisiae demonstrated lysophospholipase and lysophospholipase transacylase activities at pH 4.0. Targeted disruption of CnLYSO1 did not affect growth, melanin or capsule production by C. neoformans. Secreted lysophospholipase and transacylase activities (pH 4.0) were 50% of wild type and CnLyso1p was undetectable on Western blots. Phospholipase B activity was reduced at pH 7.0 (P<0.006) and at pH 4.0 (P=NS). The amount of secreted Plb1p (the gene product of PLB1) was also reduced. Deletion of PLB1 abolished all three secreted activities at pH 4.0 and 7.0. These results are best explained by post translational interaction, most likely the formation of a functional complex between the independently regulated gene products, CnLyso1p and CnPlb1p. PMID- 14563554 TI - Phylogenetic relationships of the Fox (Forkhead) gene family in the Bilateria. AB - The Forkhead or Fox gene family encodes putative transcription factors. There are at least four Fox genes in yeast, 16 in Drosophila melanogaster (Dm) and 42 in humans. Recently, vertebrate Fox genes have been classified into 17 groups named FoxA to FoxQ. Here, we extend this analysis to invertebrates, using available sequences from D. melanogaster, Anopheles gambiae (Ag), Caenorhabditis elegans (Ce), the sea squirt Ciona intestinalis (Ci) and amphioxus Branchiostoma floridae (Bf), from which we also cloned several Fox genes. Phylogenetic analyses lend support to the previous overall subclassification of vertebrate genes, but suggest that four subclasses (FoxJ, L, N and Q) could be further subdivided to reflect their relationships to invertebrate genes. We were unable to identify orthologs of Fox subclasses E, H, I, J, M and Q1 in D. melanogaster, A. gambiae or C. elegans, suggesting either considerable loss in ecdysozoans or the evolution of these subclasses in the deuterostome lineage. Our analyses suggest that the common ancestor of protostomes and deuterostomes had a minimum complement of 14 Fox genes. PMID- 14563555 TI - Promoter analysis of the human translation termination factor 1 gene. AB - The human translation termination factor 1 (ETF1) gene encodes a class-1 release factor, eRF1, which catalyses termination of protein synthesis at all three stop codons. In this report, we describe the functional organization of the 5'-region of the gene. Primer extension and ribonuclease protection mapping revealed three transcription start sites clustered within approximately 10 bp. DNase I hypersensitive site analysis identified five hypersensitive sites, one of which was located downstream of the initiation start sites. We used transient expression assays to define the 5'-regulating regions and in vivo and in vitro footprinting analysis to identify potential cis-acting regulatory elements. A basal promoter, spanning nucleotides -210/+117, contained no TATA box but a putative initiator element (Inr) and multiple potential Sp1/Sp3 binding sites, and thus displayed some of the features of a housekeeping gene. An additional upstream promoter containing positive and negative regulatory elements also regulated ETF1 gene expression. Real-time quantitative RT-PCR analysis showed tissue-specific expression of ETF1 transcripts in mouse tissues. Our results are suggestive of a constitutive expression of the human ETF1 gene but with possible cell- and tissue-specific regulation. PMID- 14563556 TI - ephA9, a novel avian receptor tyrosine kinase gene. AB - Protein tyrosine kinase (TK) receptors are known to play crucial roles in various aspects of normal development and in tumorigenesis. Germ cells before their colonizing to the gonads during embryogenesis are called primordial germ cells (PGCs). To identify TK genes expressed in chicken PGCs, we purified these cells from the blood of 2.5-day-old embryos, extracted the polyA(+) RNA, and subjected it to reverse transcription-polymerase chain reaction (RT-PCR) amplification with TK gene-specific primers. As a result, we identified 13 receptor TK genes and 6 non-receptor TK genes. One of the receptor TK genes appeared to be novel, and thus the full-length DNA complementary to RNA (cDNA) sequence was retrieved by the rapid cloning of cDNA ends method. Sequence analysis of this cDNA indicated that it encoded a novel TK receptor of the Eph family. The putative amino-acid sequence of this TK was 63.0% identical to that of human EphA1; therefore, we designated our novel TK as EphA9. Northern blot hybridizations indicated that ephA9 RNA transcripts were present in the kidney, lung, testis, and thymus but not in the spleen, brain, or liver. Expression of a fusion protein containing the intracellular domain of EphA9 in bacterial cells showed that this domain had TK enzymatic activity. The EphA9 species produced in Cos-1 cells transfected with an expression vector were tyrosine-phosphorylated. These data suggest that EphA9 plays its biological role(s) in various organs of chickens as an active TK. PMID- 14563557 TI - The diversity of bovine MHC class II DRB3 genes in Japanese Black, Japanese Shorthorn, Jersey and Holstein cattle in Japan. AB - We sequenced exon 2 of the major histocompatibility complex (MHC) class II DRB3 gene from 471 individuals in four different Japanese populations of cattle (201 Japanese Black, 101 Holstein, 100 Japanese Shorthorn, and 69 Jersey cattle) using a new method for sequence-based typing (SBT). We identified the 34 previously reported alleles and four novel alleles. These alleles were 80.0-100.0% identical at the nucleotide level and 77.9-100.0% identical at the amino acid level to the bovine MHC (BoLA)-DRB3 cDNA clone NR1. Among the 38 alleles, eight alleles were found in only one breed in this study. However, these alleles did not form specific clusters on a phylogenetic tree of 236-base pairs (bp) nucleotide sequences. Furthermore, these breeds exhibited similar variations with respect to average frequencies of nucleotides and amino acids, as well as synonymous and non synonymous substitutions, in all pairwise comparisons of the alleles found in this study. By contrast, analysis of the frequencies of the various BoLA-DRB3 alleles in each breed indicated that DRB3*1101 was the most frequent allele in Holstein cattle (16.8%), DRB3*4501 was the most frequent allele in Jersey cattle (18.1%), DRB3*1201 was the most frequent allele in Japanese Shorthorn cattle (16.0%) and DRB3*1001 was the most frequent allele in Japanese Black cattle (17.4%), indicating that the frequencies of alleles were differed in each breed. In addition, a population tree based on the frequency of BoLA-DRB3 alleles in each breed suggested that Holstein and Japanese Black cattle were the most closely related, and that Jersey cattle were more different from both these breeds than Japanese Shorthorns. PMID- 14563558 TI - Temperature-dependent developmental variation in lobster muscle myosin heavy chain isoforms. AB - The temperature- and developmental-regulation of myosin heavy chain (MyHC) expression and primary sequence was investigated in the abdominal musculature of developing Homarus gammarus larvae acclimated to 10, 14 and 19+/-1 degrees C. MyHC loop 1 (ATP binding) and loop 2 (actin binding) regions were sequenced and compared. The deduced amino acid sequence of MyHC loop 1 showed a development related increase in net charge from +1 to +2 between larval stages 1 and 2, which was not temperature-dependent. In post-settled stage 9 larvae, minor shifts in amino acid sequence occurred at 19 degrees C, and corresponded to a significant up-regulation of fast myosin mRNA expression. However, no temperature-specific loop 1 isoforms were detected. The deduced amino acid sequence of MyHC loop 2 was not affected by temperature, and the net charge remained +4 throughout development. These findings contrast to previous studies using the common carp, in which temperature-specific MyHC isoform genes were expressed in response to disparate thermal regimes. This raises the question as to whether arthropods do not express specific temperature isoforms but instead rely on shifts in fibre type to accommodate alterations in thermal environment. PMID- 14563559 TI - Comparison of gene representation in midguts from two phytophagous insects, Bombyx mori and Ips pini, using expressed sequence tags. AB - Midgut proteins may provide new molecular targets for insect control. This could be particularly important for some pests, such as pine bark beetles, which are difficult to control by conventional methods. Expressed sequence tags (ESTs) provide information about the activity of a particular tissue, and, in the case of pest insects, may quickly identify potential targets. We present here an EST project representing 574 tentative unique genes (TUGs) expressed in the midgut of the male pine engraver beetle, Ips pini. This tissue uses the mevalonate pathway to produce the monoterpenoid pheromone component, ipsdienol, de novo in response to juvenile hormone (JH) III. Comparison of our ESTs with those previously isolated from larval silkmoth (Bombyx mori) midguts revealed interesting similarities and differences in gene representation that correlate with the conserved and divergent functions of these two tissues. For example, seven mevalonate pathway genes were represented in the I. pini ESTs, while none were found from B. mori. This type of comparison may assist the identification of species-specific targets for future pest control strategies. PMID- 14563560 TI - Specific gene inhibition by adenovirus-mediated expression of small interfering RNA. AB - Specific gene inhibition in mammalian cells can be achieved by the use of small interfering RNA molecules (siRNA). These siRNA molecules can be chemically synthesized and transfected into cells, or directly expressed intracellularly from a plasmid DNA by the function of the cellular RNA polymerase III. We report here that the latter concept can be incorporated into an adenovirus vector to achieve specific gene inhibition in mammalian cells. As an examination of this approach, we have prepared an adenoviral vector capable of expressing siRNA molecules targeting p53 or VprBP/KIAA0800, a cellular protein that interacts with the HIV auxiliary protein Vpr. In both cases, specific reduction in the target protein level was observed after adenoviral infection. The reduction in the protein level was correlated with a specific reduction in the mRNA level. Since many cell types can be efficiently infected by adenoviruses, adenoviral vectors may serve as a useful alternative to other methods for siRNA delivery and gene inhibition, especially when the target cells are refractory to transfection by DNA or RNA. PMID- 14563562 TI - Identification and characterization of the reptilian GnRH-II gene in the leopard gecko, Eublepharis macularius, and its evolutionary considerations. AB - To elucidate the molecular phylogeny and evolution of a particular peptide, one must analyze not the limited primary amino acid sequences of the low molecular weight mature polypeptide, but rather the sequences of the corresponding precursors from various species. Of all the structural variants of gonadotropin releasing hormone (GnRH), GnRH-II (chicken GnRH-II, or cGnRH-II) is remarkably conserved without any sequence substitutions among vertebrates, but its precursor sequences vary considerably. We have identified and characterized the full-length complementary DNA (cDNA) encoding the GnRH-II precursor and determined its genomic structure, consisting of four exons and three introns, in a reptilian species, the leopard gecko Eublepharis macularius. This is the first report about the GnRH-II precursor cDNA/gene from reptiles. The deduced leopard gecko prepro GnRH-II polypeptide had the highest identities with the corresponding polypeptides of amphibians. The GnRH-II precursor mRNA was detected in more than half of the tissues and organs examined. This widespread expression is consistent with the previous findings in several species, though the roles of GnRH outside the hypothalamus-pituitary-gonadal axis remain largely unknown. Molecular phylogenetic analysis combined with sequence comparison showed that the leopard gecko is more similar to fishes and amphibians than to eutherian mammals with respect to the GnRH-II precursor sequence. These results strongly suggest that the divergence of the GnRH-II precursor sequences seen in eutherian mammals may have occurred along with amniote evolution. PMID- 14563561 TI - Expression regulation and genomic organization of human polynucleotide phosphorylase, hPNPase(old-35), a Type I interferon inducible early response gene. AB - An overlapping pathway screening (OPS) approach designed to identify and clone genes displaying parallel expression profiles as a function of induction of terminal differentiation and cellular senescence in human cells identified a novel gene old-35. Sequence and functional analysis indicates that old-35 encodes human polynucleotide phosphorylase, hPNPase(old-35). Polynucleotide phosphorylases comprise a family of phosphate dependent 3'-5' RNA exonucleases implicated in RNA regulation. Treatment of HO-1 human melanoma and additional diverse normal and tumor-derived human cell types with Type I interferon (IFN), IFN-beta or IFN-alpha, induces hPNPase(old-35) expression. To provide insights into the regulation of hPNPase(old-35), we cloned and analyzed the promoter region of this gene. These studies demonstrate that IFN-beta controls hPNPase(old 35) expression by transcriptional modulation rather than by altering mRNA stability. Transcriptional activation of hPNPase(old-35) by IFN-beta is primarily mediated by the interferon stimulatory response element (ISRE) present in its promoter. Analysis of hPNPase(old-35) expression in cell lines defective in various IFN signaling molecules confirms that hPNPase(old-35) expression is dependent upon the Janus activated kinase (JAK)/signal transducers and activators of transcription (STAT) pathway. Furthermore, gel shift analyses document that hPNPase(old-35) is a direct target of the interferon stimulated gene factor 3 (ISGF3) complex. The hPNPase(old-35) gene spans approximately 54 kb of genomic DNA and is distributed on 28 exons and 27 introns. hPNPase(old-35) maps to 2p15 2p16.1, a region implicated in hereditary nonpolyposis colorectal cancer, Carney complex, Doyne's honeycomb retinal dystrophy and several other diseases. To provide insights into PNPase function in vivo, we have also cloned the mouse PNPase(old-35) cDNA, mPNPase(old-35). Induction of hPNPase(old-35) by IFN treatment as well as during differentiation and senescence suggest that this gene may play a significant role in regulating cellular growth and that overlapping gene expression changes, also induced by IFN, may contribute to these important physiological processes. PMID- 14563563 TI - Genetic analysis of an important oxidative stress locus in the anaerobe Bacteroides fragilis. AB - The obligate anaerobe, Bacteroides fragilis, is a highly aerotolerant intestinal tract organism that has evolved a complex oxidative stress response (OSR). The redox regulator OxyR controls several OSR genes (katB, dps, and ahpC), but there is little else known about other genes it regulates. To identify additional genes in the OxyR regulon, two-dimensional gel electrophoresis was used to isolate proteins from a mutant that constitutively expresses genes in the regulon. The 28,500 Da protein thioredoxin peroxidase (Tpx) was identified. Two additional genes induced during oxidative stress were identified adjacent to tpx, a putative RNA-binding protein (rbpA) and a cytochrome-c peroxidase (ccp). Transcriptional analyses showed that tpx and rbpA were transcribed as monocistronic mRNA species or as a bicistronic operon. Transcription of tpx was induced by exposure to air or H(2)O(2) from an OxyR-dependent promoter and to a lesser extent from a second OxyR-independent promoter. Expression of the rbpA gene during oxidative stress was regulated by the OxyR-dependent tpx promoter resulting in the bicistronic tpx/rbp mRNA. The ccp gene was expressed only as a monocistronic message and induction was only observed after exposure to H(2)O(2) in an OxyR-independent manner. Disruption of the tpx operon or ccp resulted in sensitivity to the organic peroxides cumene hydroperoxide (CHP) and t-butyl hydroperoxide (TBHP) but not to H(2)O(2). This work brings the total of oxyR-controlled genes in B. fragilis to five and suggests the existence of a second peroxide response regulator that controls ccp expression. PMID- 14563564 TI - Inducible expression system and marker-linked mutagenesis approach for functional genomics of Haemophilus influenzae. AB - Complete bacterial genomic DNA sequences have allowed application of genome-scale strategies to identify essential genes. Efficient generation of conditional loss of function mutations provides a means of characterization of this class of genes. Promoter systems conferring tight regulation are particularly applicable to generating such mutations and we sought to apply this approach for the analysis of an essential gene of Haemophilus influenzae. Therefore, we developed the use of a D-xylose-inducible promoter for verification of an essential role in growth for yraM, which encodes a putative periplasmic lipoprotein, in both H. influenzae Rd and virulent type b strains. This promoter was sufficiently tightly regulated to generate conditionally viable strains by inducible expression of YraM. A second approach was used to further characterize YraM. Natural transformation and an ordered mutant collection spanning the H. influenzae genome provide the means to target any gene of interest for mutagenesis and temperature sensitive (TS) mutant isolation. This strategy was applied to generate a conditionally lethal allele of yraM. The resulting TS mutation was directly mapped to a single amino acid substitution within a motif conserved in all putative YraM orthologs and this mutation was shown to be sufficient to confer the TS phenotype. PMID- 14563565 TI - Characterization and function of kuruma shrimp lysozyme possessing lytic activity against Vibrio species. AB - Lysozyme cDNA was isolated from a kuruma shrimp, Marsupenaeus japonicus, hemocyte cDNA library. The cDNA consists of 1055 base pairs (bp) and encodes a chicken type (c-type) lysozyme with a deduced amino acid sequence of 156 residues. The kuruma shrimp lysozyme has a high identity (79.7%) with pacific white shrimp lysozyme, and low to moderate identities (33.3-43.0%) with lysozymes of insects and vertebrates. Comparisons with other c-type lysozymes from invertebrates and vertebrates showed that the two catalytic residues (Glu58 and Asp75) and the eight cysteine residue motif were completely conserved. Two novel insertion sequences were also observed in the kuruma and pacific white shrimp lysozyme amino acid sequences. Interestingly, phylogenetic analysis revealed that the kuruma shrimp lysozyme was more closely related to vertebrate c-type lysozymes. Expression of the cDNA in insect cells, using a baculovirus expression system, yielded a recombinant lysozyme with optimum activity at pH 7.5 and 50 degrees C, as evaluated by a lysoplate assay. The kuruma shrimp lysozyme displayed lytic activities against several Vibrio species and fish pathogens, including Vibrio penaeicida (a pathogenic bacteria to the kuruma shrimp) and suggested that shrimp lysozyme affects a greater variety of pathogens. PMID- 14563566 TI - Observability in strategic models of viability selection. AB - Strategic models of frequency-dependent viability selection, in terms of mathematical systems theory, are considered as a dynamic observation system. Using a general sufficient condition for observability of nonlinear systems with invariant manifold, it is studied whether, observing certain phenotypic characteristics of the population, the development of its genetic state can be recovered, at least near equilibrium. PMID- 14563567 TI - Beyond input-output computings: error-driven emergence with parallel non distributed slime mold computer. AB - The emergence derived from errors is the key importance for both novel computing and novel usage of the computer. In this paper, we propose an implementable experimental plan for the biological computing so as to elicit the emergent property of complex systems. An individual plasmodium of the true slime mold Physarum polycephalum acts in the slime mold computer. Modifying the Elementary Cellular Automaton as it entails the global synchronization problem upon the parallel computing provides the NP-complete problem solved by the slime mold computer. The possibility to solve the problem by giving neither all possible results nor explicit prescription of solution-seeking is discussed. In slime mold computing, the distributivity in the local computing logic can change dynamically, and its parallel non-distributed computing cannot be reduced into the spatial addition of multiple serial computings. The computing system based on exhaustive absence of the super-system may produce, something more than filling the vacancy. PMID- 14563568 TI - Ancient Wings: animating the evolution of butterfly wing patterns. AB - Character optimization methods can be used to reconstruct ancestral states at the internal nodes of phylogenetic trees. However, seldom are these ancestral states visualized collectively. Ancient Wings is a computer program that provides a novel method of visualizing the evolution of several morphological traits simultaneously. It allows users to visualize how the ventral hindwing pattern of 54 butterflies in the genus Bicyclus may have changed over time. By clicking on each of the nodes within the evolutionary tree, the user can see an animation of how wing size, eyespot size, and eyespot position relative the wing margin, have putatively evolved as a collective whole. Ancient Wings may be used as a pedagogical device as well as a research tool for hypothesis-generation in the fields of evolutionary, ecological, and developmental biology. PMID- 14563569 TI - Physical law not natural selection as the major determinant of biological complexity in the subcellular realm: new support for the pre-Darwinian conception of evolution by natural law. AB - Before Darwin many biologists considered organic forms to be immutable natural forms or types which like inorganic forms such as atoms or crystals are part of a changeless world order and determined by physical law. Adaptations were viewed as secondary modifications of these 'crystal like' abstract afunctional 'givens of physics.' We argue here that much of the emerging picture of biological order in the subcellular realm resembles closely the pre-Darwinian conception of nature. We point out that in the subcellular realm, between nano and micrometers, physical law necessarily plays a far more significant role in organizing matter than in the familiar 'Darwinian world' between millimeters and meters (where matter can be arranged into almost any contingent artifactual arrangement we choose, as witness Lego toys, watches or jumbo jets). Consequently, when deploying matter into complex structures in the subcellular realm the cell must necessarily make extensive use of natural forms-such as the protein and RNA folds, microtubular forms and tensegrity structures-which like atoms or crystals self-organize under the direction of physical law into what are essentially 'pre Darwinian' afunctional abstract molecular architectures in which adaptations are trivial secondary modifications of what are evidently primary givens of physics. PMID- 14563570 TI - Can we determine if the linear nature of quantum mechanics is violated by the perceptual process? AB - It now appears feasible to be able to subject to experimental test Ghirardi's proposal concerning utilizing superposed photon states to check reduction mechanisms in perceptual processes which may be governed by nonlinear evolution laws. This can be accomplished with existing techniques involving superposed single photon states and living retinal tissue mounted on microelectrode arrays. PMID- 14563571 TI - Effective dimensionality for principal component analysis of time series expression data. AB - Large-scale expression data are today measured for thousands of genes simultaneously. This development has been followed by an exploration of theoretical tools to get as much information out of these data as possible. Several groups have used principal component analysis (PCA) for this task. However, since this approach is data-driven, care must be taken in order not to analyze the noise instead of the data. As a strong warning towards uncritical use of the output from a PCA, we employ a newly developed procedure to judge the effective dimensionality of a specific data set. Although this data set is obtained during the development of rat central nervous system, our finding is a general property of noisy time series data. Based on knowledge of the noise-level for the data, we find that the effective number of dimensions that are meaningful to use in a PCA is much lower than what could be expected from the number of measurements. We attribute this fact both to effects of noise and the lack of independence of the expression levels. Finally, we explore the possibility to increase the dimensionality by performing more measurements within one time series, and conclude that this is not a fruitful approach. PMID- 14563572 TI - A randomized, double-blind, placebo-controlled trial of Ad5FGF-4 gene therapy and its effect on myocardial perfusion in patients with stable angina. AB - OBJECTIVES: The primary objective of this study was to determine whether intracoronary administration of the adenoviral gene for fibroblast growth factor (Ad5FGF-4) can improve myocardial perfusion compared with placebo. BACKGROUND: Animal studies and observational clinical studies have shown improvement in perfusion of the ischemic myocardium using genes encoding angiogenic growth factors; however, randomized, double-blind data in humans are lacking. METHODS: We performed a randomized, double-blind, placebo-controlled trial of intracoronary injection of 10(10) adenoviral particles containing a gene encoding fibroblast growth factor (Ad5FGF-4) to determine the effect on myocardial perfusion. Fifty-two patients with stable angina and reversible ischemia comprising >9% of the left ventricle on adenosine single-photon emission computed tomography (SPECT) imaging were randomized to gene therapy (n = 35) or placebo (n = 17). Clinical follow-up was performed, and 51 (98%) patients underwent a second adenosine SPECT scan after 8 weeks. RESULTS: Overall (n = 52), the mean total perfusion defect size at baseline was 32.4% of the left ventricle, with 20% reversible ischemia and 12.5% scar. At eight weeks, Ad5FGF-4 injection resulted in a significant reduction of ischemic defect size (4.2% absolute, 21% relative; p < 0.001) and placebo-treated patients had no improvement (p = 0.32). Although the change in reversible perfusion defect size between Ad5FGF-4 and placebo was not significant (4.2% vs. 1.6%, p = 0.14), when a single outlier was excluded a significant difference was observed (4.2% vs. 0.8%, p < 0.05). Ad5FGF-4 was well tolerated and did not result in any permanent adverse sequelae. CONCLUSIONS: Intracoronary injection of Ad5FGF-4 showed an encouraging trend for improved myocardial perfusion; however, further studies of therapeutic angiogenesis with Ad5FGF-4 will be necessary. PMID- 14563573 TI - The safety and efficacy of subcutaneous enoxaparin versus intravenous unfractionated heparin and tirofiban versus placebo in the treatment of acute ST segment elevation myocardial infarction patients ineligible for reperfusion (TETAMI): a randomized trial. AB - OBJECTIVES: The aims of the Safety and Efficacy of Subcutaneous Enoxaparin Versus Intravenous Unfractionated Heparin and Tirofiban Versus Placebo in the Treatment of Acute ST-Segment Elevation Myocardial Infarction Patients Ineligible for Reperfusion (TETAMI) study were to demonstrate that enoxaparin was superior to unfractionated heparin (UFH) and that tirofiban was better than placebo in patients with acute ST-segment elevation myocardial infarction (STEMI) who do not receive timely reperfusion. BACKGROUND: An optimal treatment strategy has not been identified for the many STEMI patients ineligible for acute reperfusion. METHODS: A total of 1224 patients were enrolled in 91 centers in 14 countries between July 1999 and July 2002. Patients with STEMI ineligible for reperfusion were randomized to enoxaparin, enoxaparin plus tirofiban, UFH, or UFH plus tirofiban. All patients received oral aspirin. The primary efficacy end point was the 30-day combined incidence of death, reinfarction, or recurrent angina; the primary analysis was the comparison of the pooled enoxaparin and UFH groups. RESULTS: The incidence of the primary efficacy end point was 15.7% enoxaparin versus 17.3% for UFH (odds ratio 0.89 [95% confidence interval CI = 0.66 to 1.21]) and 16.6% for tirofiban versus 16.4% for placebo (odds ratio 1.02 [95% CI 0.75 to 1.38]). The Thrombolysis In Myocardial Infarction (TIMI) major hemorrhage rate was 1.5% for enoxaparin versus 1.3% for UFH (odds ratio 1.16 [95% CI 0.44 to 3.02]) and 1.8% versus 1% for tirofiban versus placebo (odds ratio 1.82 [95% CI 0.67 to 4.95]). CONCLUSIONS: This study did not show that enoxaparin significantly reduced the 30-day incidence of death, reinfarction, and recurrent angina compared with UFH in non-reperfused STEMI patients. However, enoxaparin appears to have a similar safety and efficacy profile to UFH and may be an alternative treatment. Additional therapy with tirofiban did not appear beneficial. PMID- 14563574 TI - Optimizing outcomes in ST-segment elevation myocardial infarction. PMID- 14563575 TI - A randomized trial of polytetrafluoroethylene-membrane-covered stents compared with conventional stents in aortocoronary saphenous vein grafts. AB - We compared a conventional stent (Jostent Flex, Jomed GmbH, Rangendingen, Germany) with a polytetrafluoroethylene (PTFE)-membrane-covered stent (Jostent Stentgraft) in patients undergoing intervention of a stenosis in an obstructed vein graft. The use of stents improved results of percutaneous revascularization of obstructed vein grafts, but did not demonstrate the reduced elevated restenosis rate. In addition, long-term clinical event rate is still high compared with intervention in native vessels. Observational studies suggested that stents covered with a PTFE membrane might be associated with a low complication and restenosis rate in venous bypass grafts. This prospective multicenter study included a total of 211 patients who were randomly assigned to receive either a Flex stent or Stentgraft. The primary end point was binary restenosis rate at six months by core lab quantitative coronary angiography. Acute success and procedural events were comparable between the two groups. Restenosis rate was not significantly different between the Flex (20%) and the Stentgraft (29%) groups (p = 0.15), although there was a nonsignificant trend toward a higher late occlusion rate in the Stentgraft group (7% vs. 16%, p = 0.069) at follow-up. Likewise, after a mean observation period of 14 months, cumulative event rates (death, myocardial infarction, or target lesion revascularization) were comparable in the two groups (31% vs. 31%, p = 0.93). This controlled trial does not indicate a superiority of the PTFE-membrane covered Stentgraft compared with a conventional stent with respect to acute results, restenosis, or clinical event rates. PMID- 14563576 TI - Percutaneous treatment of saphenous vein graft disease: the ongoing challenge. PMID- 14563577 TI - Correlates of one-year survival inpatients with cardiogenic shock complicating acute myocardial infarction: angiographic findings from the SHOCK trial. AB - OBJECTIVES: The goal of this study was to describe the core laboratory angiographic findings of "SHould we emergently revascularize Occluded Coronaries for cardiogenic shocK" (SHOCK) trial participants and to determine the relationship of angiographic parameters to one-year survival. BACKGROUND: In the SHOCK trial, emergency revascularization improved one-year survival of patients with cardiogenic shock compared with initial medical stabilization including thrombolysis and intraaortic balloon counterpulsation. METHODS: Coronary angiography was performed by protocol in 147 of 152 (97%) patients in the emergency revascularization (ERV) group and by clinical selection in 100 of 150 (67%) patients in the initial medical stabilization (IMS) group. Of the other 50 IMS patients, 45 of 50 (90%) died rapidly and did not undergo angiography. RESULTS: Left ventricular ejection fraction was correlated with one-year survival in both treatment groups (p < 0.001). In the IMS group, the hazard ratio for death was 2.59 (95% confidence interval 1.47 to 4.58, p = 0.001) per diseased vessel (0/1 vs. 2 vs. 3). In the ERV group, the hazard ratio for death per diseased vessel was 1.11 (95% confidence interval 0.79 to 1.56, p = 0.559). Multivariate analysis of the angiography cohort (without regard for left ventriculogram measurements) identified initial Thrombolysis in Myocardial Infarction flow grade (p = 0.032), number of diseased vessels (for IMS patients only, p = 0.024), and culprit vessel (p = 0.004) as independent correlates of one year survival, even after adjustment for key clinical factors. In the smaller cohort with left ventricular ejection fraction measured (n = 97), ejection fraction and culprit vessel remained independently correlated with survival. CONCLUSIONS: For patients in cardiogenic shock, left ventricular function and culprit vessel were independent correlates of one-year survival. PMID- 14563578 TI - Percutaneous coronary intervention for cardiogenic shock in the SHOCK trial. AB - OBJECTIVES: We examined the clinical, angiographic, and procedural characteristics determining survival after percutaneous coronary intervention (PCI) for cardiogenic shock. BACKGROUND: The SHOCK (SHould we emergently revascularize Occluded coronaries for Cardiogenic shocK?) trial prospectively enrolled patients with shock complicating acute myocardial infarction (MI). Patients were randomized to a strategy of early revascularization or initial medical stabilization. METHODS: Patients randomized to early revascularization underwent PCI or bypass surgery on the basis of predefined clinical criteria. Patients randomized to early revascularization who underwent PCI and had angiographic films available for analysis are the subject of this report (n = 82). RESULTS: The median time from MI to PCI was 11 h. The majority of patients had occluded culprit arteries (Thrombolysis In Myocardial Infarction [TIMI] grade 0 or 1 flow in 62%) and multivessel disease (81%). One-year mortality in PCI patients was 50%. Mortality was 39% if PCI was successful but 85% if unsuccessful (p < 0.001). Mortality was 38% if TIMI flow grade 3 was achieved, 55% with TIMI grade 2 flow, and 100% with TIMI grade 0 or 1 flow (p < 0.001). Mortality was 67% if severe mitral regurgitation was documented. Independent correlates of mortality were as follows: increasing age (p < 0.001), lower systolic blood pressure (p = 0.009), increasing time from randomization to PCI (p = 0.019), lower post-PCI TIMI flow (0/1 vs. 2/3) (p < 0.001), and multivessel PCI (p = 0.040). CONCLUSIONS: Restoration of coronary blood flow is a major predictor of survival in cardiogenic shock. Benefit appears to extend beyond the generally accepted 12-h post-infarction window. Surgery should be considered in shock patients with severe mitral insufficiency or multivessel disease not amenable to relatively complete percutaneous revascularization. PMID- 14563579 TI - Cardiogenic shock: is there light at the end of the tunnel? PMID- 14563580 TI - Effect of activated protein C on plasma plasminogen activator inhibitor activity in patients with acute myocardial infarction treated with alteplase: comparison with unfractionated heparin. AB - OBJECTIVES: We examined whether activated protein C (APC) is an effective conjunctive therapy to thrombolysis in patients with ST-segment-elevated acute myocardial infarction (AMl). BACKGROUND: Activated protein C possesses both systemic anticoagulant and anti-inflammatory properties. It has been also shown to enhance fibrinolysis by inhibiting plasminogen activator inhibitor (PAI) activity in vitro. METHODS: After successful thrombolysis with alteplase, study patients were assigned to receive one of the two conjunctive therapies for 48 h intravenously: human plasma-derived APC at 0.06 mg/kg per day (APC group, n = 9) or unfractionated heparin at 100 to 400 U/kg per day, adjusted to maintain an activated partial thromboplastin time at 1.5 to 2 times of the control level (heparin group, n = 10). RESULTS: Adverse events, including reocclusion of the recanalized infarct-related coronary artery and major or minor hemorrhagic complications, occurred more frequently in the heparin group (4 of 10 cases) than in the APC group (none of 9 cases) (p = 0.033). In the heparin group, plasma PAI activity (IU/ml, median value [range]) was increased continuously from 8 to 24 h after thrombolysis and peaked at 24 h (30.9 [11.3 to 38.5]); on the other hand, it was not increased in the APC group at 24 h after thrombolysis (11.3 [0.0 to 31.0], p < 0.01 vs. heparin group). CONCLUSIONS: Administration of APC suppressed increasing of plasma PAI activity observed after thrombolysis in patients with AMI. The effect of APC could be more eligible, compared with heparin, as a conjunctive regimen to thrombolysis in AMI patients. PMID- 14563581 TI - Intracoronary thrombectomy improves myocardial reperfusion in patients undergoing direct angioplasty for acute myocardial infarction. AB - OBJECTIVES: We sought to evaluate the effects of mechanical thrombectomy on myocardial reperfusion during direct angioplasty for acute myocardial infarction (AMI). BACKGROUND: Embolization of thrombus and plaque debris may occur during direct angioplasty for AMI. This may lead to distal vessel or side branch occlusion and to obstructions in the microvascular system, resulting in impaired myocardial reperfusion. Mechanical thrombectomy is used to reduce distal embolization. METHODS: Ninety-two patients with AMI and angiographic evidence of intraluminal thrombus were randomized to either intracoronary thrombectomy followed by stenting or to a conventional strategy of stenting. Thrombectomy was performed using the X-Sizer catheter (EndiCOR Inc., San Clemente, California). Myocardial reperfusion was assessed by myocardial blush and ST resolution. RESULTS: Postprocedure Thrombolysis in Myocardial Infarction-3 flow was not different between groups (93.5% vs. 95.7%, p = 0.39). Myocardial blush-3 was observed in 71.7% of patients undergoing thrombectomy and in 36.9% of patients undergoing conventional strategy (p = 0.006). ST-segment resolution >or=50% occurred more often in patients undergoing thrombectomy (82.6% vs. 52.2%, p = 0.001). By multivariate analysis, adjunctive thrombectomy was an independent predictor of blush-3 (odds ratio, 3.27; 95% confidence interval, 1.06 to 10.05; p = 0.039). CONCLUSIONS: Intracoronary thrombectomy as adjunct to stenting during direct angioplasty for AMI improves myocardial reperfusion as assessed by myocardial blush and ST resolution. PMID- 14563582 TI - Interventional therapy for acute myocardial infarction: respect the microvasculature. PMID- 14563583 TI - Mortality risk conferred by small elevations of creatine kinase-MB isoenzyme after percutaneous coronary intervention. AB - OBJECTIVES: The aim of this study was to assess whether small creatine kinase-MB isoenzyme (CK-MB) elevations after percutaneous coronary intervention (PCI) affect the subsequent mortality risk. BACKGROUND: Several studies have evaluated the relationship of CK-MB levels after PCI with the subsequent risk of death. While there is consensus that elevations exceeding 5 times the upper limit of normal increase mortality significantly, there is uncertainty about the exact clinical impact of smaller CK-MB elevations. METHODS: We performed a meta analysis of seven studies with CK-MB measurements and survival outcomes on 23230 subjects who underwent PCI. Data were combined with random effects models. RESULTS: Mean follow-up was 6 to 34 months per study. By random effects, 19% (95% confidence interval [CI], 16% to 23%) had one- to five-fold CK-MB elevations, while only 6% (95% CI, 5% to 9%) had >5-fold elevations. Compared with subjects with normal CK-MB, there was a dose-response relationship with relative risks for death being 1.5 (95% CI, 1.2 to 1.8, no between-study heterogeneity) with one- to three-fold CK-MB elevations, 1.8 (95% CI, 1.4 to 2.4, no between-study heterogeneity) with three- to five-fold CK-MB elevations, and 3.1 (95% CI, 2.3 to 4.2, borderline between-study heterogeneity) with over five-fold CK-MB elevations (p < 0.001 for all). CONCLUSIONS: Any increase in CK-MB after PCI is associated with a small, but statistically and clinically significant, increase in the subsequent risk of death. PMID- 14563584 TI - Periprocedural myocardial infarction and mortality: causality versus association. PMID- 14563585 TI - Addition of paclitaxel to contrast media prevents restenosis after coronary stent implantation. AB - OBJECTIVES: The present study was designed to test the efficacy of paclitaxel added to the contrast agent iopromide in the prevention of restenosis. BACKGROUND: Contrast media adhere to the coronary vessel wall for some seconds after injection. Such a layer of contrast agent could serve as a matrix for antiproliferative drugs. METHODS: Thirty-four stents were implanted into the left anterior descending and circumflex coronary arteries of 17 pigs, using a 1.2:1.0 overstretch ratio. The unsupplemented contrast agent iopromide-370 was used as a control; the treatment groups were treated with 80 ml intracoronary iopromide plus either 100 or 200 mumol/l paclitaxel, or 80 ml intravenous iopromide plus 200 mumol/l paclitaxel. Quantitative angiography and histomorphometry were used to assess comparable baseline parameters between the treatment groups. RESULTS: A short time incubation (3 min) almost completely inhibited vascular smooth muscle cell proliferation, sustained for up to 12 days. Whereas intravenous paclitaxel had no effect, intracoronary application of paclitaxel reduced the diameter stenosis from 55 +/- 13% to 29 +/- 18% and 13 +/- 12%. Late lumen loss dropped from 1.94 +/- 0.35 mm under the control condition to 1.19 +/- 0.55 mm with 100 mumol/l paclitaxel and to 0.82 +/- 0.54 mm with 200 mumol/l paclitaxel. Histomorphometry revealed a corresponding dose-dependent reduction of the neointimal area and restenosis by intracoronary iopromide paclitaxel. Assessment of left ventricular function and myocardial histology revealed no adverse effects of intracoronary paclitaxel application. CONCLUSIONS: This study provides evidence that intracoronary application of a taxane dissolved in a contrast medium profoundly inhibits in-stent restenosis. This novel, widely feasible approach may be suited for the prevention of restenosis in a broad spectrum of interventional treatment regimens. PMID- 14563586 TI - The elderly: health status benefits and recovery of function one year after coronary artery bypass surgery. AB - OBJECTIVES: The purpose of this study was to describe the health status (symptoms, function, and quality of life) changes of elderly patients undergoing coronary artery bypass grafting (CABG) and compare these to younger patients. BACKGROUND: Despite increasing use of CABG in the elderly, few data exist about elderly patients' health status benefits from CABG. METHODS: A total of 690 consecutive patients (n = 156, >75 years of age; n = 534, T polymorphism of the apolipoprotein E gene in low-risk men, the -1171/5A- >6A polymorphism of the stromelysin-1 gene in low-risk women, the 1019C-->T polymorphism of the connexin 37 gene in high-risk men, and the 3932T-->C polymorphism of the apolipoprotein E gene in high-risk women were significantly associated with CAD. A stepwise forward selection procedure revealed that the effects of these polymorphisms on CAD were statistically independent of age or conventional risk factors. CONCLUSIONS: Genotyping of these polymorphisms may prove informative for assessment of the genetic risk of CAD in low- or high-risk men or women. PMID- 14563590 TI - Predictors of late development of heart failure in stable survivors of myocardial infarction: the CARE study. AB - OBJECTIVES: We sought to determine the predictors of heart failure (HF) development in long-term survivors of myocardial infarction (MI). BACKGROUND: Modern strategies of acute MI care have resulted in an increasing proportion of survivors at heightened risk of future non-fatal events, including HF. METHODS: We assessed the risk of developing HF in 3860 stable MI patients without a previous history of HF, who were enrolled in the Cholesterol And Recurrent Events (CARE) trial a median of 10 months post MI. Baseline characteristics of patients who did or did not develop HF during the five years of observation were assessed. RESULTS: A total of 243 patients (6.3%) developed HF in a linear pattern at a rate of 1.3%/year. Heart failure development markedly increased the risk of death (hazard ratio 10.2, 95% confidence interval 7.7 to 13.5). Fifty-seven patients (23.5%) who developed HF had a recurrent MI between enrollment and the onset of HF, increasing the risk fivefold. The most important predictors of HF were age and left ventricular ejection fraction. Other predictors included diabetes, history of hypertension, previous MI, and baseline heart rate. Moderate exercise three or more times per week was independently associated with a 30% lower risk of HF. CONCLUSIONS: Heart failure post MI occurs in a time-dependent fashion, which is usually not a direct consequence of a detectable interim MI. Patients who experience late-onset HF have a 10-fold increased risk of death compared with other MI survivors. Baseline characteristics can risk stratify patients at high risk of subsequent HF. PMID- 14563591 TI - Cardiac resynchronization therapy for the treatment of heart failure in patients with intraventricular conduction delay and malignant ventricular tachyarrhythmias. AB - OBJECTIVES: This study was conducted to assess the safety and effectiveness of cardiac resynchronization therapy (CRT) when combined with an implantable cardioverter defibrillator (ICD). BACKGROUND: Long-term outcome of CRT was measured in patients with symptomatic heart failure (HF), intraventricular conduction delay, and malignant ventricular tachyarrhythmias (ventricular tachycardia/ventricular fibrillation [VT/VF]) requiring therapy from an ICD. METHODS: Patients (n = 490) were implanted with a device capable of providing both CRT and ICD therapy and randomized to CRT (n = 245) or control (no CRT, n = 245) for up to six months. The primary end point was progression of HF, defined as all-cause mortality, hospitalization for HF, and VT/VF requiring device intervention. Secondary end points included peak oxygen consumption (VO(2)), 6 min walk (6 MW), New York Heart Association (NYHA) class, quality of life (QOL), and echocardiographic analysis. RESULTS: A 15% reduction in HF progression was observed, but this was statistically insignificant (p = 0.35). The CRT, however, significantly improved peak VO(2) (0.8 ml/kg/min vs. 0.0 ml/kg/min, p = 0.030) and 6 MW (35 m vs. 15 m, p = 0.043). Changes in NYHA class (p = 0.10) and QOL (p = 0.40) were not statistically significant. The CRT demonstrated significant reductions in ventricular dimensions (left ventricular internal diameter in diastole = -3.4 mm vs. -0.3 mm, p < 0.001 and left ventricular internal diameter in systole = -4.0 mm vs. -0.7 mm, p < 0.001) and improvement in left ventricular ejection fraction (5.1% vs. 2.8%, p = 0.020). A subgroup of patients with advanced HF (NYHA class III/IV) consistently demonstrated improvement across all functional status end points. CONCLUSIONS: The CRT improved functional status in patients indicated for an ICD who also have symptomatic HF and intraventricular conduction delay. PMID- 14563592 TI - The points for pacing. PMID- 14563593 TI - Time interval between onset of mitral inflow and onset of early diastolic velocity by tissue Doppler: a novel index of left ventricular relaxation: experimental studies and clinical application. AB - OBJECTIVES: The goal of this study was to examine the diagnostic utility of the time to onset of early (Ea) diastolic velocity of the mitral annulus by tissue Doppler (TD) in comparison with the time to onset of mitral inflow (T(Ea-E)) for the assessment of left ventricular (LV) relaxation. BACKGROUND: Tissue Doppler imaging of the mitral annulus provides useful information about myocardial function. So far, studies have focused on the measurement of peak Ea, but have not evaluated the diagnostic utility of the time to onset of Ea. METHODS: Simultaneous left heart catheterization and Doppler echocardiography (DE) were performed in 10 dogs. Left atrial pressures and LV volumes and pressures were measured before and after constriction of the circumflex (cx) coronary artery. The delay in Ea was next examined in 60 consecutive patients, undergoing simultaneous right heart catheterization and DE. Furthermore, (T(Ea-E)) was used to predict filling pressures in a prospective group of 33 patients. RESULTS: In canine studies, significant prolongation in the time interval (T(Ea-E)) was noted after cx constriction, which had a significant relation with tau (tau) (r = 0.93, p < 0.01). In human studies, Ea was significantly delayed in patients with impaired relaxation and pseudonormal LV filling in comparison with age-matched controls. In the prospective group, pulmonary capillary wedge pressure (PCWP) derived as: PCWP(Doppler) = LV(end-systolic pressure) x e(-IVRT/(T(Ea-E))), where IVRT is isovolumetric relaxation time; PCWP(Doppler) related well to PCWP(catheter) (r = 0.84, p < 0.001). CONCLUSIONS: T(Ea-E) is a useful novel index of LV relaxation. It can be used to identify patients with diastolic dysfunction and predict PCWP. PMID- 14563594 TI - The return of cardiac time intervals: the phoenix is rising. PMID- 14563595 TI - Acute beta-blockade reduces the extent and severity of myocardial perfusion defects with dipyridamole Tc-99m sestamibi SPECT imaging. AB - OBJECTIVES: The goal of this study was to examine the effect of acute beta blockade on dipyridamole Tc-99m sestamibi myocardial perfusion imaging (DMPI). BACKGROUND: Studies suggest that antianginal drugs may reduce the presence and severity of myocardial perfusion defects with dipyridamole stress. However, there are no data regarding specific drugs. METHODS: Patients with catheterization proven coronary artery disease (CAD) were enrolled in this prospective, double blind, placebo-controlled study and randomly assigned to DMPI after placebo, low dose metoprolol (up to 10 mg), and high-dose metoprolol (up to 20 mg). Patients underwent one Tc-99m sestamibi study at rest on a separate day. The interval between DMPI studies was A at intron 4, 686+1G>A at intron 6, and 1244+2T>G at intron 10 (numbered from the A of the initiation Met codon), and we confirmed that 2 of these mutations produced aberrantly spliced messenger RNAs (mRNAs). The remaining 4 mutations were missense mutations: R193W, I673F, C908Y, and R1123C. In expression experiments using HeLa cells, all mutants showed no or a marginal secretion of ADAMTS13. Taken together with the findings in our recent report we determined the responsible mutations in a total of 7 Japanese patients with USS with a uniform clinical picture of severe neonatal hyperbilirubinemia, and in their family members, based on ADAMTS13 gene analysis. Of these patients, 2 were homozygotes and 5 were compound heterozygotes. The parents of one homozygote were related (cousins), while those of the other were not. Molecular models of the metalloprotease, fifth domain of thrombospondin 1 (Tsp1-5), and Tsp1-8 domains of ADAMTS13 suggest that the missense mutations could cause structural defects in the mutants. PMID- 14563639 TI - E mu-BRD2 transgenic mice develop B-cell lymphoma and leukemia. AB - Transgenic mice with lymphoid-restricted overexpression of the double bromodomain protein bromodomain-containing 2 (Brd2) develop splenic B-cell lymphoma and, upon transplantation, B-cell leukemia with leukemic infiltrates in liver and lung. Brd2 is a nuclear-localized transcription factor kinase that is most closely related to TATA box binding protein-associated factor, 250 kDa (TAF(II)250) and the Drosophila developmental protein female sterile homeotic. Constitutive expression of BRD2 in the lymphoid compartment increases cyclin A transcription, "priming" transgenic B cells for proliferation. Mice stochastically develop an aggressive B-cell lymphoma with the features of B-1 cells, including CD5 and surface IgM expression. The B-cell lymphoma is monoclonal for immunoglobulin gene rearrangement and is phenotypically stable. The lymphoblasts are very large and express a transcriptome that is similar to human non-Hodgkin lymphomas. Both a wild-type BRD2 transgene and a kinase-null point mutant drive lymphomagenesis; therefore we propose that, rather than kinase activity, Brd2-mediated recruitment of E2 promoter binding factors (E2Fs) and a specific histone acetyltransferase to the cyclin A promoter by both types of transgene is a mechanistic basis for neoplasia. This report is the first to describe a transgenic mouse model for constitutive expression of a protein with more than one bromodomain. PMID- 14563641 TI - Regulation of MHC class II expression in human T-cell malignancies. AB - Expression of major histocompatibility complex (MHC) class II molecules in human activated T cells is under normal circumstances regulated exclusively by the CIITA-PIII subtype of the class II transactivator (CIITA). In this study, we show that the absence of MHC class II expression in leukemic T cells was due to a lack of expression of CIITA, whereas in T-lymphoma cells, expression of CIITA correlated with expression of MHC class II. Interestingly, activation of a CIITA promoter (P)III-reporter construct was not affected in leukemic T cells. This revealed that the absence of endogenous CIITA expression was not caused by a lack of transcription factors critical for CIITA-PIII activation but suggests the involvement of an epigenetic silencing mechanism. Subsequent analysis showed that the lack of human leukocyte antigen-DR (HLA-DR) expression correlated with hypermethylation of CIITA-PIII in leukemic T-cell lines and in primary T-cell acute lymphoblastic leukemia (T-ALL) and a T-cell prolymphocytic leukemia (T PLL). Treatment of leukemic T-cell lines with a demethylation agent showed re expression of CIITA-PIII and HLA-DRA. Furthermore, in vitro methylation of CIITA PIII and subsequent assessment of CIITA-PIII activity in Jurkat leukemic T cells resulted in reduction of constitutive and CREB-1 (cyclic adenosine monophosphate [cAMP]-response element binding protein 1)-induced promoter activity. Together, these results argue for an important role of DNA hyper-methylation in the control of CIITA expression in leukemic T cells. PMID- 14563642 TI - Nucleophosmin-anaplastic lymphoma kinase of anaplastic large-cell lymphoma recruits, activates, and uses pp60c-src to mediate its mitogenicity. AB - Anaplastic large-cell lymphomas (ALCLs) are lymphomas of T or null phenotype often associated with a chromosomal translocation, t(2;5)(p23;q35). This translocation leads to the expression of a hybrid protein consisting of the N terminal portion of nucleophosmin (NPM) and the intracellular domain of the anaplastic lymphoma kinase (ALK). NPM-ALK possesses a constitutive tyrosine kinase activity responsible for its oncogenic property through activation of downstream effectors such as phospholipase C gamma (PLC-gamma) and the type IA phosphoinositide 3-kinase. Here, we show that the Src-kinases, particularly pp60(c-src), associate with and are activated by NPM-ALK expression in various cells, and in cell lines established from patients with ALCL. The kinase activity and the tyrosine 418 of NPM-ALK are required for its association with Src kinases. Y418F mutation of NPM-ALK impaired its association with Src-kinases and strongly reduced the proliferation rate of Ba/F3 cells. In agreement, Src-kinase inhibitors or pp60(c-src) siRNA significantly decreased the proliferation rate of NPM-ALK-positive ALCL cell lines. Moreover, using active or inactive forms of pp60(c-src) and NPM-ALK, we provide evidence that NPM-ALK is a potential substrate of pp60(c-src). Overall, our data place Src-kinases as new important downstream effectors of NPM-ALK and as attractive potential therapeutic targets for new ALCL treatment. PMID- 14563643 TI - LPAM (alpha 4 beta 7 integrin) is an important homing integrin on alloreactive T cells in the development of intestinal graft-versus-host disease. AB - Lymphocyte Peyer patch adhesion molecule (LPAM) or alpha(4)beta(7) integrin is expressed on lymphocytes and is responsible for T-cell homing into gut-associated lymphoid tissues through its binding to mucosal addressin cell adhesion molecule (MAdCAM), which is present on high endothelial venules of mucosal lymphoid organs. We found in murine allogeneic bone marrow transplantation (BMT) models that recipients of alpha(4)beta(7)(-) donor T cells had significantly less graft versus-host disease (GVHD) morbidity and mortality compared with recipients of alpha(4)beta(7)(+) donor T cells. A kinetic posttransplantation analysis of lymphocytes in the intestines and mesenteric lymph nodes demonstrated a delayed invasion of lower numbers of alpha(4)beta(7)(+) T cells in recipients of alpha(4)beta(7)(-) T cells compared with recipients of alpha(4)beta(7)(+) T cells. Histopathologic analysis of GVHD target organs revealed that recipients of alpha(4)beta(7)(-) T cells developed less GVHD of the intestines and liver, whereas there was no difference in cutaneous and thymic GVHD between recipients of alpha(4)beta(7)(-) or alpha(4)beta(7)(+) T cells. Finally, we found that in vivo GVT activity of alpha(4)beta(7)(-) donor T cells was preserved. We conclude that the alpha(4)beta(7) integrin is important for the invasion of alloreactive donor T cells into the gut and the subsequent development of intestinal GVHD and overall GVHD morbidity and mortality. PMID- 14563644 TI - Direct killing of Epstein-Barr virus (EBV)-infected B cells by CD4 T cells directed against the EBV lytic protein BHRF1. AB - Due to their low frequency, CD4 T-cell responses to Epstein-Barr virus (EBV) lytic antigens are, so far, poorly characterized. Human peptide major histocompatibility complex (MHC) class II multimers provide a means to detect and characterize such rare T cells. Along a screening of T-cell responses to lytic or latent EBV antigens within peripheral blood leukocyte (PBL)- or synovial-derived CD4 T-cell lines, we identified an human leukocyte antigen-DR*0401 (HLA-DR*0401) restricted epitope derived from BHRF1 (BamHI fragment H rightward open reading frame 1), a viral protein produced during the early stages of the lytic cycle. We show here that T-cell responses to this particular BHRF1 epitope are shared by most EBV-infected DR*0401(+) individuals, as BHRF1-specific CD4 T cells could be sorted out from all the DRB*0401 T-cell lines analyzed, using magnetic beads coated with recombinant BHRF1/DR*0401 complexes. Sorting with these peptide MHC class II multimers was very efficient, as the yield of recovery of BHRF1-specific T cells was nearly 100%. Functional analysis of a large number of clones responding to BHRF1/DR*0401 demonstrated their cytolytic action against autologous and allogeneic DR*0401(+) EBV-transformed B-lymphoblastoid cell lines (B-LCLs), with 40% to 80% killing efficiency and potent interferon gamma production, thus suggesting that this CD4 T-cell population contributes to the control of EBV replication. B-LCL lysis by these T-cell clones was DR*0401 dependent, EBV dependent, and was not merely due to bystander killing. Taken together, these data provide the first demonstration that a lytic antigen can induce a direct cytolytic response against EBV-infected cells. PMID- 14563645 TI - Mechanism of protein sorting during erythroblast enucleation: role of cytoskeletal connectivity. AB - During erythroblast enucleation, nuclei surrounded by plasma membrane separate from erythroblast cytoplasm. A key aspect of this process is sorting of erythroblast plasma membrane components to reticulocytes and expelled nuclei. Although it is known that cytoskeletal elements actin and spectrin partition to reticulocytes, little is understood about molecular mechanisms governing plasma membrane protein sorting. We chose glycophorin A (GPA) as a model integral protein to begin investigating protein-sorting mechanisms. Using immunofluorescence microscopy and Western blotting we found that GPA sorted predominantly to reticulocytes. We hypothesized that the degree of skeletal linkage might control the sorting pattern of transmembrane proteins. To explore this hypothesis, we quantified the extent of GPA association to the cytoskeleton in erythroblasts, young reticulocytes, and mature erythrocytes using fluorescence imaged microdeformation (FIMD) and observed that GPA underwent dramatic reorganization during terminal differentiation. We discovered that GPA was more connected to the membrane cytoskeleton, either directly or indirectly, in erythroblasts and young reticulocytes than in mature cells. We conclude that skeletal protein association can regulate protein sorting during enucleation. Further, we suggest that the enhanced rigidity of reticulocyte membranes observed in earlier investigations results, at least in part, from increased connectivity of GPA with the spectrin-based skeleton. PMID- 14563646 TI - Platelet receptor interplay regulates collagen-induced thrombus formation in flowing human blood. AB - The platelet glycoproteins (GPs) Ib, integrin alpha(2)beta(1), and GPVI are considered central to thrombus formation. Recently, their relative importance has been re-evaluated based on data from murine knockout models. To examine their relationship during human thrombus formation on collagen type I fibers at high shear (1000 s(-1)), we tested a novel antibody against GPVI, an immunoglobulin single-chain variable fragment, 10B12, together with specific antagonists for GPIb alpha (12G1 Fab(2)) and alpha(2)beta(1) (6F1 mAb or GFOGER-GPP peptide). GPVI was found to be crucial for aggregate formation, Ca(2+) signaling, and phosphatidylserine (PS) exposure, but not for primary adhesion, even with more than 97% receptor blockade. Inhibiting alpha(2)beta(1) revealed its involvement in regulating Ca(2+) signaling, PS exposure, and aggregate size. Both GPIb alpha and alpha(2)beta(1) contributed to primary adhesion, showing overlapping function. The coinhibition of receptors revealed synergism in thrombus formation: the coinhibition of adenosine diphosphate (ADP) receptors with collagen receptors further decreased adhesion and aggregation, and, crucially, the complete eradication of thrombus formation required the coinhibition of GPVI with either GPIb alpha or alpha(2)beta(1). In summary, human platelet deposition on collagen depends on the concerted interplay of several receptors: GPIb in synergy with alpha(2)beta(1) mediating primary adhesion, reinforced by activation through GPVI, which further regulates the thrombus formation. PMID- 14563647 TI - Functional endothelial cells derived from rhesus monkey embryonic stem cells. AB - We have used rhesus monkey embryonic stem (ES) cells to study endothelial cell development. Rhesus ES cells (R366.4 cell line) exposed to medium containing vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF), and epidermal growth factor (EGF) assumed a relatively uniform endothelial cell morphology and could be propagated and expanded with a consistent phenotype and normal karyotype. When placed in Matrigel, these rhesus ES cell-derived endothelial cells (RESDECs) formed capillary-like structures characteristic of endothelial cells. Immunohistochemical and flow cytometric analysis of RESDECs showed that they take up acetylated low-density lipoprotein (LDL), express CD146, von Willebrand factor, and the integrin alpha v beta 3, and bind the lectin ulex europaeus agglutinin-1. These cells also express the VEGF receptor Flk-1 and secrete VEGF. When introduced in a Matrigel plug implanted subcutaneously in mice, RESDECs formed intact vessels and recruited new endothelial cell growth. In vivo function was demonstrated by coinjection of RESDECs with murine tumor cells subcutaneously into immunocompromised adult mice. RESDECs injected alone did not form measurable tumors. Tumor cells grew more rapidly and had increased vascularization when coinjected with the RESDECs. Immunohistochemical staining demonstrated that the RESDECs participated in forming the tumor neovasculature. RESDECs provide a novel means to examine the mechanisms of endothelial cell development, and may open up new therapeutic strategies. PMID- 14563648 TI - Dynamic regulation of Src-family kinases by CD45 in B cells. AB - CD45 is a key protein tyrosine phosphatase regulating Src-family protein tyrosine kinases (Src-PTKs) in lymphocytes; precisely how it exerts its effect remains controversial, however. We previously demonstrated that CD45 negatively regulates Lyn in the WEHI-231 B-cell line. Here we show that negative regulation by CD45 is physiologically significant in B cells and that some CD45 is constitutively associated with glycolipid-enriched microdomains (GEMs), where it inhibits Src PTKs by dephosphorylating both the negative and the positive regulatory sites. Upon B-cell receptor (BCR) ligation, however, CD45 dissociates from GEMs within 30 seconds, inducing phosphorylation of 2 regulatory sites and activation of Src PTKs, but subsequently reassociates with the GEMs within 15 minutes. Disruption of GEMs with methyl-beta-cyclodextrin results in abrogation of BCR-induced apoptosis in WEHI-231 cells, suggesting GEMs are critical to signals leading to the fate determination. We propose that the primary function of CD45 is inhibition of Src-PTKs and that the level of Src-PTK activation and the B-cell fate are determined in part by dynamic behavior of CD45 with respect to GEMs. PMID- 14563649 TI - Estrogen receptor-alpha polymorphisms and angiographic outcome after coronary artery stenting. AB - OBJECTIVE: Because of the receptor-mediated antiproliferative effects of estradiol on vascular smooth muscle cells, our study aimed at identifying a role of PvuII and XbaI polymorphisms of the alpha-estrogen receptor (alphaER) gene in the occurrence of restenosis after coronary stent implantation (in-stent restenosis [ISR]). METHODS AND RESULTS: In 858 patients (148 women), 955 lesions were treated with stent implantation, and the PvuII C/T and XbaI G/A polymorphisms of the alphaER gene were determined. Quantitative angiography was performed before and after stenting and at 6-month follow-up. The allelic frequencies were similar between sexes (C/T allele, 0.43/0.57 and 0.44/0.56; P=0.9; G/A allele, 0.35/0.65 and 0.38/0.62; P=0.8; in women and men, respectively). A significantly higher ISR rate in women than in men homozygous for the T-allele of the PvuII polymorphism (42.6% versus 26.9%, P=0.03) or the G allele of the XbaI polymorphism (41.2% versus 19.4%, P=0.04) was observed. At multivariate analysis, T/T genotype was the only independent predictor of ISR in women but not in men (odds ratio, 1.5; 95% CI, 1.0 to 2.1; P=0.03). XbaI polymorphism was no longer associated with ISR in both sexes. CONCLUSIONS: Women homozygous for the T-allele of the PvuII polymorphism of the alphaER gene treated with coronary stent implantation have a higher risk of ISR than men. PMID- 14563650 TI - Genetic ablation of caveolin-1 confers protection against atherosclerosis. AB - OBJECTIVE: The development of atherosclerosis is a process characterized by the accumulation of lipids in the form of modified lipoproteins in the subendothelial space. This initiating step is followed by the subsequent recruitment and proliferation of other cell types, including monocytes/macrophages and smooth muscle cells. Here, we evaluate the potential role of caveolae membrane domains in the pathogenesis of atherosclerosis by using apolipoprotein E-deficient (ApoE /-) mice as a model system. METHODS AND RESULTS: Caveolin-1 (Cav-1) is a principal structural protein component of caveolae membrane domains. To directly assess the in vivo role of caveolae and Cav-1 in atherosclerosis, we interbred Cav-1-/- mice with ApoE-/- mice. Interestingly, loss of Cav-1 resulted in a dramatic >2-fold increase in non-HDL plasma cholesterol levels in the ApoE-/- background. However, despite this hypercholesterolemia, we found that loss of Cav 1 gene expression was clearly protective against the development of aortic atheromas, with up to an approximately 70% reduction in atherosclerotic lesion area. Mechanistically, we demonstrated that loss of Cav-1 resulted in the dramatic downregulation of certain proatherogenic molecules, namely, CD36 and vascular cell adhesion molecule-1. CONCLUSIONS: Taken together, our results indicate that loss of Cav-1 can counteract the detrimental effects of atherogenic lipoproteins. Thus, Cav-1 is a novel target for drug development in the pharmacologic prevention of atheroma formation. Our current data also provide the first molecular genetic evidence to support the hypothesis that caveolar transcytosis of modified lipoproteins (from the blood to the sub-endothelial space) is a critical initiating step in atherosclerosis. PMID- 14563651 TI - Alcohol consumption and carotid atherosclerosis in older adults: the Cardiovascular Health Study. AB - OBJECTIVE: The association of alcohol use with atherosclerosis is inconsistent in previous studies. METHODS AND RESULTS: For the Cardiovascular Health Study, 5888 adults aged 65 years and older underwent a standardized interview and examination. They reported beer, wine, and liquor use individually and underwent B-mode ultrasonography to determine internal and common carotid intima-media thickness (IMT). We compared composite carotid IMT values cross-sectionally using linear regression to adjust for demographic and clinical characteristics. Among 4247 participants free of cardiovascular disease, consumers of 1 to 6 drinks per week had 0.07+/-0.04-mm lower composite IMT and consumers of 14 or more drinks per week had 0.07+/-0.05-mm higher IMT than abstainers (P quadratic trend=0.02). We found similar relationships using internal and common carotid thickness measures and among men and women. The higher IMT associated with heavier alcohol use was particularly strong among 1592 participants with confirmed cardiovascular disease (0.24+/-0.09 mm greater than abstainers). Controlling for HDL cholesterol levels reduced the effect on composite IMT among consumers of 1 to 6 drinks per week by 22%. CONCLUSIONS: Relative to older adults who abstain from alcohol, consumption of 1 to 6 drinks per week had an inverse association with carotid atherosclerosis whereas consumption of 14 or more drinks had a positive association. PMID- 14563652 TI - Association of the Toll-like receptor 4 gene Asp299Gly polymorphism with acute coronary events. AB - OBJECTIVE: Atherosclerosis is a chronic inflammatory disease of the blood vessels. Toll-like receptor 4 (TLR4) is a transmembrane receptor that is involved in mediating inflammatory responses to bacterial endotoxin and other ligands. The aim of this study was to search for an association between a common functional polymorphism of TLR4--Asp299Gly--and acute coronary syndrome. METHODS AND RESULTS: We conducted a case-control study of 183 patients with acute coronary syndromes and 216 controls. We screened the TLR4 gene for the Asp299Gly polymorphism using a 5' fluorogenic assay. The 299Gly allele was associated with a decreased risk of acute coronary events independently of standard coronary risk factors. The adjusted odds ratio associated with this allele was 0.41 (95% CI, 0.18 to 0.95; P=0.037). In controls, TLR4 heterozygosity was also associated with a significant decrease in plasma fibrinogen and soluble vascular cellular adhesion molecule-1 levels (P<0.01). CONCLUSIONS: These results, which must be confirmed by a prospective longitudinal study, provide evidence of an association between the Asp299Gly polymorphism of the human TLR4 receptor and acute coronary syndromes. They confirm the previously reported involvement of TLR4 in carotid and femoral artery atherosclerosis. PMID- 14563653 TI - Antiinflammatory roles of peroxisome proliferator-activated receptor gamma in human alveolar macrophages. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) is a ligand activated transcriptional factor belonging to the nuclear receptor superfamily. PPARgamma, which is predominantly expressed in adipose tissue, plays a major regulatory role in glucose metabolism and adipogenesis. Interestingly, recent studies have demonstrated PPARgamma expression in monocytes/macrophages and its antiinflammatory activities. However, it is unclear whether alveolar macrophages (AMs) express functional PPARgamma. The present study was conducted to investigate the expression of PPARgamma by AMs and to elucidate its functional role. Using reverse transcription-polymerase chain reaction and Western blotting, we demonstrated the strong expression of PPARs messenger RNA and protein in freshly isolated human AMs. Ligands of PPARgamma, 15-deoxy delta(12,14)prostaglandin J2, and troglitazone significantly decreased LPS induced tumor necrosis factor-alpha production by AMs. These ligands markedly upregulated the expression of CD36, a scavenger receptor that mediates the phagocytosis of apoptotic neutrophils. Indeed, ligand-treated AMs ingested a significantly higher number of apoptotic neutrophils than untreated AMs. These data indicate that PPARgamma expressed by AMs play an antiinflammatory role through inhibiting cytokine production and increasing their CD36 expression together with the enhanced phagocytosis of apoptotic neutrophils, which is an essential process for the resolution of inflammation. This suggests the potential therapeutic application of PPARgamma ligands in inflammatory disorders of the lung. PMID- 14563654 TI - Efficacy of alendronate in adults with cystic fibrosis with low bone density. AB - As adults with cystic fibrosis (CF) have enjoyed incremental increases in longevity over the last few decades, they have also been suffering from low bone density and its clinical manifestations, fractures and kyphosis. We conducted a placebo-controlled, randomized, double-blinded trial of alendronate (10 mg/day orally) (n = 24) compared with placebo (n = 24) for 1 year in 48 patients to improve bone mineral density at the spine as the primary endpoint. All patients received 800 IU of cholecalciferol and 1,000 mg of calcium carbonate. Both groups were similar in age, sex, CF mutations, bone density T scores, renal function, and body mass index at study onset. The alendronate-treated patients gained (mean +/- SD) 4.9 +/- 3.0% and 2.8 +/- 3.2% bone density after 1 year versus placebo, which lost (mean +/- SD) 1.8 +/- 4.0% and 0.7 +/- 4.7%, in spine and femur bone density, respectively (p < or = 0.001 for the spine; p = 0.003 for the femur). Urine N-telopeptide, a bone resorption marker, levels declined in the treatment group more than in the control group (p = 0.002), consistent with the known antiresorptive effects of bisphosphonates. Alendronate was more effective than placebo in improving spine and femur bone mineral density and is a promising agent for the long-term prevention and management of bone disease in patients with CF. PMID- 14563655 TI - Role of sensory nerve peptides rather than mast cell histamine in paclitaxel hypersensitivity. AB - Paclitaxel is one of the most extensively used anticancer agents, however, its use is often limited by severe hypersensitivity reactions, including respiratory distress, bronchospasm, and hypotension, which can occur despite premedication with dexamethasone and histamine H1 and H2 antagonists. The present study was designed to determine the mechanisms of paclitaxel hypersensitivity. In rats, paclitaxel (15 mg/kg, intravenously) caused a marked increase in pulmonary vascular permeability and edema. PaO2 decreased, whereas PaCO2 increased, transiently after paclitaxel injection. The paclitaxel-induced pulmonary vascular hyperpermeability was blocked by dexamethasone but not by histamine H1 or H2 antagonists. Paclitaxel increased the vascular permeability in lungs of mast cell deficient rats Ws/Ws(-/-) to almost the similar extent as that elicited in wild type rats. On the other hand, the paclitaxel-induced pulmonary vascular hyperpermeability was reversed by sensory denervation with capsaicin or pretreatment with LY303870 and SR48968, NK1 and NK2 antagonists, respectively. Consistent with these findings, a marked elevation of sensory neuropeptides such as substance P, neurokinin A, and calcitonin gene-related peptide was observed in rat bronchoalveolar lavage fluid after paclitaxel injection. These findings suggest that sensory nerves rather than mast cells are implicated in the etiology of paclitaxel hypersensitivity. PMID- 14563656 TI - Cathelicidin peptide sheep myeloid antimicrobial peptide-29 prevents endotoxin induced mortality in rat models of septic shock. AB - The present study was designed to investigate the antiendotoxin activity and therapeutic efficacy of sheep myeloid antimicrobial peptide (SMAP)-29, a cathelicidin-derived peptide. The in vitro ability of SMAP-29 to bind LPS from Escherichia coli 0111:B4 was determined using a sensitive limulus chromogenic assay. Two rat models of septic shock were performed: (1) rats were injected intraperitoneally with 1 mg E. coli 0111:B4 LPS and (2) intraabdominal sepsis was induced via cecal ligation and single puncture. All animals were randomized to receive parenterally isotonic sodium chloride solution, 1 mg/kg SMAP-29, 1 mg/kg polymyxin B or 20 mg/kg imipenem. The main outcome measures were: abdominal exudate and plasma bacterial growth, plasma endotoxin and tumor necrosis factor alpha concentrations, and lethality. The in vitro study showed that SMAP-29 completely inhibited the LPS procoagulant activity at approximately 10 microM peptide concentration. The in vivo experiments showed that all compounds reduced the lethality when compared with control animals. SMAP-29 achieved a substantial decrease in endotoxin and tumor necrosis factor-alpha plasma concentrations when compared with imipenem and saline treatment and exhibited a slightly lower antimicrobial activity than imipenem. No statistically significant differences were noted between SMAP-29 and polymyxin B. SMAP-29, because of its double antiendotoxin and antimicrobial activities, could be an interesting compound for septic shock treatment. PMID- 14563657 TI - Bovine lactoferrin has a nitric oxide-dependent hypotensive effect in rats. AB - Lactoferrin (LF) is a multifunctional protein that is found in milk, neutrophils, and other biological fluids. Under inflammatory conditions, LF production is increased in the periphery by neutrophils. However, the cardiovascular function of LF is still unknown. In the present study, we investigated the effect of bovine LF (BLF) on the mean blood pressure (MBP) and heart rate (HR) in urethane anesthetized rats and the vascular function of BLF in the rat thoracic aorta. Intravenous injection of BLF produced dose-dependent decreases in MBP but did not affect HR, while the opioid agonist morphine decreased both MBP and HR. The hypotensive effect of BLF was not altered by naloxone methiodide, which cannot pass through the blood-brain barrier, but was significantly reduced by naloxone hydrochloride, which does pass through the blood-brain barrier. BLF-induced hypotension was completely blocked by the NO synthase inhibitor NG-nitro-l arginine methyl ester (l-NAME) but not by the inactive enantiomer of l-NAME, NG nitro-d-arginine methyl ester (d-NAME). BLF-induced hypotension was not altered by the muscarinic ACh receptor antagonist atropine or the cyclooxygenase inhibitor diclofenac. BLF produced relaxation in endothelium-intact but not endothelium-denuded aortic rings precontracted with phenylephrine. The relaxation evoked by BLF was completely blocked by l-NAME but not by d-NAME or the ATP sensitive potassium channel blocker glibenclamide. These results suggest that BLF causes hypotension via an endothelium-dependent vasodilation that is strongly mediated by NO production and that BLF-induced hypotension also may be mediated by the central opioidergic system. PMID- 14563658 TI - LiCl-induced flavor avoidance compared between rats and mice using a nondeprivation protocol. AB - The present studies examine some parameters involved in flavor avoidance learning, using LiCl to induce malaise, in a novel nondeprivation protocol that allows direct comparison between rats and mice. The procedure involves daily presentation of a gelatin dessert that contains carbohydrate (Polycose) and a distinctive food flavor. Regular chow is additionally available at all times. Both rats and mice showed robust intakes of these gels with little change of gram intake as concentration of Polycose was varied in the range 2-30%; at the highest concentration, the caloric yield was approximately 7% of normal daily intake in both species. Rats that were injected on three occasions with LiCl (0.75 meq/kg) 1 h after consumption of a flavored gel formed a complete and sustained conditioned flavor avoidance (CFA). In a two-flavor discrimination protocol, in which a second flavor was followed by injections of saline, rats showed complete avoidance of the LiCl-paired flavor and partial avoidance of the saline-paired flavor. Mice injected on three occasions with LiCl (6 meq/kg) 1 h after intake of a flavored gel formed a partial CFA; a more complete CFA was formed when there was no delay between removal of the flavor and the injection. Using this no-delay protocol, mice, like rats, showed avoidance of a saline-paired flavor in a two flavor discrimination protocol, and the CFA was strong when the dose of LiCl was reduced to that used in rats (0.75 meq/kg). In comparable protocols, mice thus are able to form complete CFAs using low doses of LiCl that are comparable to CFAs observed in rats, but the interval between flavor and sickness over which associative learning can occur may be shorter in mice. PMID- 14563659 TI - Ablation of renin-expressing juxtaglomerular cells results in a distinct kidney phenotype. AB - Renin-expressing cells are peculiar in that they act as differentiated cells, producing the hormone renin, while they also seem to act as progenitors for other renal cell types. As such, they may have functions independent of their ability to generate renin/angiotensin. To test this hypothesis, we ablated renin expressing cells during development by placing diphtheria toxin A chain (DTA) under control of the Ren1d mouse renin promoter by homologous recombination in a two-renin gene strain (Ren2 and Ren1d). Renin-expressing cells are essentially absent from kidneys in homozygotes (DTA/DTA) which, unlike wild-type mice, are unable to recruit renin-expressing cells when homeostasis is threatened. In contrast, renin staining in the submandibular gland (SMG), which expresses mainly Ren2, is normal. Homozygous mice survive normally, but the kidneys are small and have morphological abnormalities: 25% of the glomeruli are hyperplastic or atrophic, tubules are dilated and atrophic, and areas of undifferentiated cells exist near the atrophic glomeruli and tubules. However, in contrast to the very abnormal renal vessels found when renin-angiotensin system genes are deleted, the kidney vessels in homozygotes have normal wall thickness and no decrease in lumen size. Homozygotes have severely reduced kidney and plasma renin concentrations and females have reduced blood pressure. Homozygotes have elevated blood urea nitrogen and potassium levels, which are suggestive of altered renal function. We conclude that renin cells per se are necessary for the morphological integrity of the kidney and may have a role in maintenance of normal kidney function. PMID- 14563660 TI - Excitatory amino acid receptors in the dorsomedial hypothalamus mediate prostaglandin-evoked thermogenesis in brown adipose tissue. AB - We determined whether the dorsomedial hypothalamus (DMH) plays a role in the thermogenic, metabolic, and cardiovascular effects evoked by centrally administered PGE2. Microinjection of PGE2 (170 pmol/60 nl) into the medial preoptic area of the hypothalamus in urethane-chloralose-anesthetized, artificially ventilated rats increased brown adipose tissue (BAT) sympathetic nerve activity (SNA; +207 +/- 18% of control), BAT temperature (1.5 +/- 0.2 degrees C), expired CO2 (0.9 +/- 0.1%), heart rate (HR; 106 +/- 12 beats/min), and mean arterial pressure (22 +/- 4 mmHg). Within 5 min of subsequent bilateral microinjections of the GABAA receptor agonist muscimol (120 pmol.60 nl-1.side-1) or the ionotropic excitatory amino acid antagonist kynurenate (6 nmol.60 nl 1.side-1) into the DMH, the PGE2-evoked increases were, respectively, attenuated by 91 +/- 3% and 108 +/- 7% for BAT SNA, by 73 +/- 12% and 102 +/- 28% for BAT temperature, by 100 +/- 4% and 125 +/- 21% for expired CO2, by 72 +/- 11% and 70 +/- 16% for HR, and by 84 +/- 19% and 113 +/- 16% for mean arterial pressure. Microinjections outside the DMH within the dorsal hypothalamic area adjacent to the mamillothalamic tracts or within the ventromedial hypothalamus were less effective for attenuating the PGE2-evoked thermogenic, metabolic, and cardiovascular responses. These results demonstrate that activation of excitatory amino acid receptors within the DMH is necessary for the thermogenic, metabolic, and cardiovascular responses evoked by microinjection of PGE2 into the medial preoptic area. PMID- 14563661 TI - New technique for measurement of left ventricular pressure in conscious mice. AB - Concern about the effects of anesthesia on physiological measurements led us to develop methodology to assess left ventricular (LV) pressure in conscious mice. Polyethylene-50 tubing filled with heparinized saline was implanted in the LV cavity through its apex via an abdominal approach and exteriorized to the back of the animal. This surgery was done under anesthesia with either an intraperitoneal injection of ketamine (80 mg/kg) and xylazine (5 mg/kg) (K+X) in 11 mice or isoflurane (ISF; 1.5 vol%) by inhalation in 14 mice. Postoperatively, mice were trained daily to lie quietly head first in a plastic cone. LV pressure, the first derivative of LV pressure (dP/dt), and heart rate (HR) in the conscious state were compared between the two groups at 3 days and 1 wk after recovery from surgery using a 1.4-Fr Millar catheter inserted into the LV through the tubing, with the mice lying quietly in the plastic cone. Acutely during anesthesia, K+X decreased HR (from 698 to 298 beats/min), LV systolic pressure (from 107 to 65 mmHg), and maximal dP/dt (dP/dt(max)) (from 15,724 to 4,445 mmHg/s), all P < 0.01. Similar but less marked negative chronotropic and inotropic effects were seen with ISF. HR and dP/dt(max) were decreased significantly in K+X mice 3 days after surgery compared with those anesthetized with ISF (655 vs. 711 beats/min, P < 0.05; 14,448 vs. 18,048 mmHg/s, P < 0.001) but increased to the same level as in ISF mice 1 wk after surgery. In ISF mice, recovery of function occurred rapidly and there were no differences in LV variables between 3 days and 1 wk. LV pressure and dP/dt can be measured in conscious mice with a micromanometer catheter inserted through tubing implanted permanently in the LV apex. Anesthesia with either K+X or, to a lesser extent, ISF, depressed LV function acutely. This depression of function persisted for 3 days after surgery with K+X (but not ISF) and did not recover completely until 1 wk postanesthesia. PMID- 14563662 TI - Interstitial purine metabolites in hearts with LV remodeling. AB - The myocardial ATP concentration is significantly decreased in failing hearts, which may be related to the progressive loss of the myocardial total adenine nucleotide pool. The total myocardial interstitial purine metabolites (IPM) in the dialysate of interstitial fluid could reflect the tissue ATP depletion. In rats, postmyocardial infarction (MI) left ventricular (LV) remodeling was induced by ligation of the coronary artery. Cardiac microdialysis was employed to assess changes of IPM in response to graded beta-adrenergic stimulation with isoproterenol (Iso) in myocardium of hearts with post-MI LV remodeling (MI group) or hearts with sham operation (sham group). The dialysate samples were analyzed for adenosine, inosine, hypoxanthine, xanthine, and uric acid. LV volume was greater in the MI group (2.2 +/- 0.2 ml/kg) compared with the sham group (1.3 +/- 0.2 ml/kg, P < 0.05). Infarct size was 28 +/- 4%. The baseline dialysate level of uric acid was higher in the MI group (18.9 +/- 3.4 micromol) compared with the sham group (4.6 +/- 0.7 micromol, P < 0.01). During and after Iso infusion, the dialysate levels of adenosine, xanthine, and uric acid were all significantly higher in the MI group. Thus the level of IPM is increased in hearts with postinfarction LV remodeling both at baseline and during Iso infusion. These results suggest that the decreased myocardial ATP level in hearts with post-MI LV remodeling may be caused by the chronic depletion of the total adenine nucleotide pool. PMID- 14563663 TI - Interaction between prostanoids and nitric oxide in regulation of systemic, pulmonary, and coronary vascular tone in exercising swine. AB - Prostacyclin and nitric oxide (NO) are produced by the endothelium in response to physical forces such as shear stress. Consequently, both NO and prostacyclin may increase during exercise and contribute to metabolic vasodilation. Conversely, NO has been hypothesized to inhibit prostacyclin production. We therefore investigated the effect of cyclooxygenase (COX) inhibition on exercise-induced vasodilation of the porcine systemic, pulmonary, and coronary beds before and after inhibition of NO production. Swine were studied at rest and during treadmill exercise at 1-5 km/h, before and after COX inhibition with indomethacin (10 mg/kg iv), and in the absence and presence of NO synthase inhibition with N(omega)-nitro-l-arginine (l-NNA; 20 mg/kg iv). COX inhibition produced systemic vasoconstriction at rest, which waned during exercise. The systemic vasoconstriction by COX inhibition was enhanced after l-NNA, particularly at rest. In the coronary circulation, COX inhibition also resulted in vasoconstriction at rest and during exercise. However, vasoconstriction was not modified by pretreatment with l-NNA. In contrast, COX inhibition had no effect on the pulmonary circulation, either at rest or during exercise. Moreover, a prostanoid influence in the pulmonary circulation could not be detected after l NNA. In conclusion, endogenous prostanoids contribute importantly to systemic and coronary tone in awake swine at rest but are not mandatory for exercise-induced vasodilation in these beds. Endogenous prostanoids are not mandatory for the regulation of pulmonary resistance vessel tone. Finally, NO blunts the contribution of prostanoids to vascular tone regulation in the systemic but not in the coronary and pulmonary beds. PMID- 14563665 TI - Carbon monoxide activates KCa channels in newborn arteriole smooth muscle cells by increasing apparent Ca2+ sensitivity of alpha-subunits. AB - Carbon monoxide (CO) is a gaseous vasodilator produced by many cell types, including endothelial and smooth muscle cells. The goal of the present study was to investigate signaling mechanisms responsible for CO activation of large conductance Ca(2+)-activated K(+) (K(Ca)) channels in newborn porcine cerebral arteriole smooth muscle cells. In intact cells at 0 mV, CO (3 microM) or CO released from dimanganese decacarbonyl (10 microM), a novel light-activated CO donor, increased K(Ca) channel activity 4.9- or 3.5-fold, respectively. K(Ca) channel activation by CO was not blocked by 1-H-[1,2,4]oxadiazolo[4,3 a]quinoxalin-1-one (25 microM), a soluble guanylyl cyclase inhibitor. In inside out patches at 0 mV, CO shifted the Ca(2+) concentration-response curve for K(Ca) channels leftward and decreased the apparent dissociation constant for Ca(2+) from 31 to 24 microM. Western blotting data suggested that the low Ca(2+) sensitivity of newborn K(Ca) channels may be due to a reduced beta-subunit-to alpha-subunit ratio. CO activation of K(Ca) channels was Ca(2+) dependent. CO increased open probability 3.7-fold with 10 microM free Ca(2+) at the cytosolic membrane surface but only 1.1-fold with 300 nM Ca(2+). CO left shifted the current-voltage relationship of cslo-alpha currents expressed in HEK-293 cells, increasing currents 2.2-fold at +50 mV. In summary, data suggest that in newborn arteriole smooth muscle cells, CO activates low-affinity K(Ca) channels via a direct effect on the alpha-subunit that increases apparent Ca(2+) sensitivity. The optimal tuning by CO of the micromolar Ca(2+) sensitivity of K(Ca) channels will lead to preferential activation by signaling modalities, such as Ca(2+) sparks, which elevate the subsarcolemmal Ca(2+) concentration within this range. PMID- 14563664 TI - Phosphoinositide 3-kinase regulates excitation-contraction coupling in neonatal cardiomyocytes. AB - The phosphoinositide 3-kinase (PI3K) inhibitor LY-294002 decreased steady-state contraction in neonatal rat ventricular myocytes (NRVM). To determine whether the effect on steady-state contraction could be due to decreased intracellular Ca(2+) content, Ca(2+) content was assessed with fluorescent plate reader analysis by using the caffeine-releasable Ca(2+) stores as an index of sarcoplasmic reticulum (SR) Ca(2+) content. Caffeine-releasable Ca(2+) content was diminished in a dose dependent manner with LY-294002, suggesting that the decrease in steady-state contraction was due to diminished intracellular Ca(2+) content. Activation of the L-type Ca(2+) channel by BAY K 8644 was attenuated by LY-294002, suggesting the effect of LY-294002 is to reduce Ca(2+) influx at this channel. To investigate whether additional proteins involved in excitation-contraction (EC) coupling are likewise regulated by PI3K activity, the effects of compounds acting at sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA2a), the ryanodine receptor, and the Na/Ca exchanger (NCX) were compared with LY-294002. Inhibition of SERCA2a by thapsigargin increased basal Ca(2+) levels in contrast to LY-294002, indicating that SERCA2a activity is sustained in the presence of LY-294002. Ryanodine decreased SR Ca(2+) content. The additive effect with coadministration of LY 294002 could be attributed to a decrease in Ca(2+) influx at the L-type Ca(2+) channel. The NCX inhibitor Ni(2+) was used to investigate whether the decrease in intracellular Ca(2+) content with LY-294002 could be due to inhibition of the NCX reverse-mode activity. The minimal effect of LY-294002 with Ni(2+) suggests that the primary effect of LY-294002 on EC coupling occurs through inhibition of PI3K mediated L-type Ca(2+) channel activity. PMID- 14563666 TI - Effect of bronchial artery blood flow on cardiopulmonary bypass-induced lung injury. AB - Cardiovascular surgery requiring cardiopulmonary bypass (CPB) is frequently complicated by postoperative lung injury. Bronchial artery (BA) blood flow has been hypothesized to attenuate this injury. The purpose of the present study was to determine the effect of BA blood flow on CPB-induced lung injury in anesthetized pigs. In eight pigs (BA ligated) the BA was ligated, whereas in six pigs (BA patent) the BA was identified but left intact. Warm (37 degrees C) CPB was then performed in all pigs with complete occlusion of the pulmonary artery and deflated lungs to maximize lung injury. BA ligation significantly exacerbated nearly all aspects of pulmonary function beginning at 5 min post-CPB. At 25 min, BA-ligated pigs had a lower arterial Po(2) at a fraction of inspired oxygen of 1.0 (52 +/- 5 vs. 312 +/- 58 mmHg) and greater peak tracheal pressure (39 +/- 6 vs. 15 +/- 4 mmHg), pulmonary vascular resistance (11 +/- 1 vs. 6 +/- 1 mmHg x l( 1) x min), plasma TNF-alpha (1.2 +/- 0.60 vs. 0.59 +/- 0.092 ng/ml), extravascular lung water (11.7 +/- 1.2 vs. 7.7 +/- 0.5 ml/g blood-free dry weight), and pulmonary vascular protein permeability, as assessed by a decreased reflection coefficient for albumin (sigma(alb); 0.53 +/- 0.1 vs. 0.82 +/- 0.05). There was a negative correlation (R = 0.95, P < 0.001) between sigma(alb) and the 25-min plasma TNF-alpha concentration. These results suggest that a severe decrease in BA blood flow during and after warm CPB causes increased pulmonary vascular permeability, edema formation, cytokine production, and severe arterial hypoxemia secondary to intrapulmonary shunt. PMID- 14563667 TI - A new model of pacing in the mouse intestine. AB - A simple model of pacing in mouse intestine to longitudinal (LM) as well as circular muscle (CM) has been developed. Undissected segments of LM or CM from mouse ileum or jejunum were prepared to record contractions, nerve functions were inhibited, and regular spontaneous contractions were recorded. These had the properties expected of interstitial cells of Cajal (ICC) paced contractions: ileum slower than jejunum, inhibited but not abolished by nicardipine, reduced in frequency by cyclopiazonic acid, abolished by Ca(2+)-free media, and high temperature dependence (Q10 approximately 2.6-3.2). Nicardipine significantly reduced the pacing frequency in LM and CM. Intestinal segments from W/W(V) mice had few irregular contractions in CM but had regular contractions in LM. Other differences were found between LM and CM that suggest that the control of pacing of LM differed from pacing of CM. Moreover, both LM and CM segments in wild-type and W/W(V) and after cyclopiazonic acid responded to electrical pacing (50 V/cm, 50 or 100 ms) at 1 pulse per second. Temperature <26 degrees C inhibited electrically paced contractions in CM. These findings suggest that the current models of ICC pacing need to be modified to apply to intact segments of mouse intestine. PMID- 14563668 TI - Transforming growth factor-alpha directly augments histidine decarboxylase and vesicular monoamine transporter 2 production in rat enterochromaffin-like cells. AB - For the production and vesicle storage of histamine, Enterochromaffin-like (ECL) cells express histidine decarboxylase (HDC) and vesicular monoamine transporter 2 (VMAT2). Although HDC and VMAT2 show dynamic changes during gastric ulcer healing, the control system of their expression has not been fully investigated. In the present study, we investigated the effect of transforming growth factor alpha (TGF-alpha) and proinflammatory cytokines on HDC and VMAT2 expression in rat ECL cells. Time course changes in the expression of TGF-alpha during the healing of acetic acid-induced ulcers were studied. EGF receptor (EGFR) expression was also examined in ECL cells, whereas the direct effects of TGF alpha and proinflammatory cytokines on HDC and VMAT2 expression in ECL cells were investigated using in vivo and in vitro models. During the process of ulcer healing, expression of TGF-alpha mRNA was markedly augmented. Furthermore, EGFR was identified in isolated ECL cells. TGF-alpha stimulated HDC and VMAT2 mRNA expression and protein production and also increased histamine release from ECL cells. Selective EGFR tyrosine kinase inhibitor tyrphostin AG1478 almost completely inhibited HDC and VMAT2 gene expression induced by TGF-alpha in vivo and in vitro. During gastric mucosal injury, TGF-alpha was found to stimulate ECL cell functions by increasing HDC and VMAT2 expression. PMID- 14563669 TI - Immunomagnetic enrichment of interstitial cells of Cajal. AB - Disruptions of networks of interstitial cells of Cajal (ICC), gastrointestinal pacemakers and mediators of neurotransmission, can lead to disordered phasic contractions and peristalsis by reducing and uncoupling electrical slow waves. However, detailed analysis of the ICC network behavior has been hampered by their scarcity, limited accessibility in intact tissues, and contamination with other cell types in culture. Our goal was to develop a simple technique to purify ICC from murine gastrointestinal muscles for functional studies. We identified ICC in live small intestinal muscles or primary cell cultures by Kit immunoreactivity using fluorescent antibodies. Because this technique also labels resident macrophages nonspecifically, parallel studies were performed in which nonfluorescent Kit antibodies and macrophages labeled with fluorescent dextran were used for subtractive analysis of ICC. In both groups, Kit-positive cells were tagged with superparamagnetic antibodies and sorted on magnetic columns. Efficacy was assessed by flow cytometry. ICC enrichment from primary cultures and freshly dissociated tissues was approximately 63-fold and approximately 8-fold, respectively. Unlike the cells derived directly from tissues, cells sorted from cultures frequently yielded extensive, nearly homogenous ICC networks on reseeding. Monitoring oscillations in mitochondrial Ca(2+) or membrane potential by imaging revealed spontaneous rhythmicity in these networks. Cells that did not bind to the columns yielded cultures that were depleted of ICC and dominated by smooth muscle cells. In conclusion, immunomagnetic sorting of primary cultures of ICC results in relatively homogenous, functional ICC networks. This technique is less suitable for obtaining ICC from freshly dispersed cells. PMID- 14563670 TI - Calcium source diversity in feline lower esophageal sphincter circular and sling muscle. AB - Within muscular equivalents of cat lower esophageal sphincter (LES), the circular muscle develops greater spontaneous tone, whereas the sling muscle is more responsive to cholinergic stimulation. Smooth muscle contraction involves a combination of calcium release from stores and of calcium entry via several pathways. We hypothesized that there are differences in the sources of Ca(2+) used for contraction in sling and circular muscles and that these differences could contribute to functional asymmetry observed within LES. Contraction of muscle strips from circular and sling regions of LES was assessed in the presence of TTX. In Ca(2+)-free Krebs, tone was inhibited to a greater degree in circular than sling muscle. L-type Ca(2+) channel blockade with nifedipine or verapamil inhibited tone in LES circular but not sling muscle. Sarcoplasmic reticulum (SR) Ca(2+)-ATPase inhibitor cyclopiazonic acid (CPA) caused greater increase in tone in sling than in circular muscle. The phospholipase C inhibitor U-73122 and the SR inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)] receptor blocker 2 aminoethoxydiphenyl borate (2-APB) inhibited tone in circular and sling muscles, demonstrating that continuous release of Ca(2+) from Ins(1,4,5)P(3)-sensitive stores is important in tone generation in both muscles. In Ca(2+)-free Krebs, ACh induced contractions (AChC) were inhibited to a greater degree in sling than circular muscles. However, nifedipine and verapamil greatly inhibited AChC in the circular but not sling muscle. Depletion of SR Ca(2+) stores with CPA or inhibition of Ins(1,4,5)P(3)-mediated store release with either U-73122 or 2-APB inhibited AChC in both muscles. We demonstrate that LES circular and sling muscles 1) use intracellular and extracellular Ca(2+) sources to different degrees in the generation of spontaneous tone and AChC and 2) use different Ca(2+) entry pathways. These differences hold the potential for selective modulation of LES tone in health and disease. PMID- 14563671 TI - Functional importance of ICAM-1 in the mechanism of neutrophil-induced liver injury in bile duct-ligated mice. AB - Cholestasis-induced liver injury during bile duct obstruction causes an acute inflammatory response. To further characterize the mechanisms underlying the neutrophil-induced cell damage in the bile duct ligation (BDL) model, we performed experiments using wild-type (WT) and ICAM-1-deficient mice. After BDL for 3 days, increased ICAM-1 expression was observed along sinusoids, along portal veins, and on hepatocytes in livers of WT animals. Neutrophils accumulated in sinusoids [358 +/- 44 neutrophils/20 high-power fields (HPF)] and >50% extravasated into the parenchymal tissue. Plasma alanine transaminase (ALT) levels increased by 23-fold, and severe liver cell necrosis (47 +/- 11% of total cells) was observed. Chlorotyrosine-protein adducts (a marker for neutrophil derived hypochlorous acid) and 4-hydroxynonenal adducts (a lipid peroxidation product) were detected in these livers. Neutrophils also accumulated in the portal venules and extravasated into the portal tracts. However, no evidence for chlorotyrosine or 4-hydroxynonenal protein adducts was detected in portal tracts. ICAM-1-deficient mice showed 67% reduction in plasma ALT levels and 83% reduction in necrosis after BDL compared with WT animals. The total number of neutrophils in the liver was reduced (126 +/- 25/20 HPF), and 85% of these leukocytes remained in sinusoids. Moreover, these livers showed minimal staining for chlorotyrosine and 4-hydroxynonenal adducts, indicating a substantially reduced oxidant stress and a diminished cytokine response. Thus neutrophils relevant for the aggravation of acute cholestatic liver injury in BDL mice accumulate in hepatic sinusoids, extravasate into the tissue dependent on ICAM-1, and cause cell damage involving reactive oxygen formation. PMID- 14563672 TI - Antioxidant activity of nitro derivative of aspirin against ischemia-reperfusion in hamster cheek pouch microcirculation. AB - Aspirin that has been chemically combined with a nitric oxide (NO) donor (NCX 4016) has been shown to inhibit cyclooxygenase and prostaglandin generation while maintaining the inhibitory effects of aspirin. The possible role of reactive oxygen species (ROS) in the action of NCX-4016 in ischemia-reperfusion (I/R) has not been studied. Furthermore, we were interested in comparing the effects of a conventional NO donor [2,2'-hydroxynitrosohydrazino-bis-etanamine (DETA/NO)] and NCX-4016 at the microvascular level in the hamster cheek pouch visualized by using an intravital fluorescent microscopy technique. Microvascular injury was assessed by measuring diameter change, the perfused capillary length (PCL), and leukocyte adhesion. Animals were treated with NCX-4016 (100 mg/kg or 30 mg.kg( 1).day(-1) for 5 days po) or DETA-NO (0.5 mg/kg). Mean arterial blood pressure increased slightly but significantly after NCX-4016 treatment. During 5- and 15 min reperfusion, lipid peroxides in the systemic blood increased by 72 and 89% vs. baseline, respectively, and were still higher than in basal conditions after 30-min reperfusion in the I/R group. Pretreatment with NCX-4016 maintained ROS at normal levels; increased arteriolar diameter, blood flow, and PCL; and decreased leukocyte adhesion (P < 0.05). DETA-NO decreased ROS during 30-min reperfusion; however, later there was a significant increase during reperfusion. DETA-NO decreased leukocyte adhesion (P < 0.05) but microvascular permeability increased after 30 min of reperfusion. In conclusion, NCX-4016 attenuates oxidative stress and prevents arteriolar constriction during I/R, whereas DETA-NO increases lipid peroxides in the systemic blood and permeability after reperfusion. PMID- 14563673 TI - Prevention of TNF-alpha-induced apoptosis in polyamine-depleted IEC-6 cells is mediated through the activation of ERK1/2. AB - It has been documented that polyamines play a critical role in the regulation of apoptosis in intestinal epithelial cells. We have recently reported that protection from TNF-alpha/cycloheximide (CHX)-induced apoptosis in epithelial cells depleted of polyamines is mediated through the inactivation of a proapoptotic mediator, JNK. In this study, we addressed the involvement of the MAPK pathway in the regulation of apoptosis after polyamine depletion of IEC-6 cells. Polyamine depletion by alpha-difluromethylornithine (DFMO) resulted in the sustained activation of ERK in response to TNF-alpha/CHX treatment. Pretreatment of polyamine-depleted IEC-6 cells with a cell membrane-permeable MEK1/2 inhibitor, U-0126, significantly inhibited TNF-alpha/CHX-induced ERK phosphorylation and significantly increased DNA fragmentation, JNK activity, and caspase-3 activity in response to TNF-alpha/CHX. Moreover, the dose dependency of U-0126-mediated inhibition of TNF-alpha/ CHX-induced ERK phosphorylation correlated with the reversal of the antiapoptotic effect of DFMO. IEC-6 cells expressing constitutively active MEK1 had decreased TNF-alpha/CHX-induced JNK phosphorylation and were significantly protected from apoptosis. Conversely, a dominant-negative MEK1 resulted in high basal activation of JNK, cytochrome c release, and spontaneous apoptosis. Polyamine depletion of the dominant-negative MEK1 cells did not prevent JNK activation or cytochrome c release and failed to confer protection from both TNF-alpha/CHX and camptothecin-induced apoptosis. Finally, expression of a dominant-negative mutant of JNK significantly protected IEC-6 cells from TNF-alpha/CHX-induced apoptosis. These data indicate that polyamine depletion results in the activation of ERK, which inhibits JNK activation and protects cells from apoptosis. PMID- 14563674 TI - Inducible antisense RNA targeting amino acid transporter ATB0/ASCT2 elicits apoptosis in human hepatoma cells. AB - Amino acid transporter B(0)/ASC transporter 2 (ATB(0)/ASCT2) is responsible for most glutamine uptake in human hepatoma cells. Because this transporter is not expressed in normal hepatocytes, we hypothesized that its expression is necessary for growth of human liver cancer cells. To test this hypothesis, Sloan Kettering hepatoma (SK-Hep) cells were stably transfected with an inducible 1.3-kb ATB(0)/ASCT2 antisense RNA expression plasmid under the transcriptional control of mifepristone, a synthetic steroid. Induced antisense RNA expression in monolayer cultures decreased ATB(0)/ASCT2 mRNA levels by 73% and glutamine transport rates by 65% compared with controls after 24 h, leading to a 98% decrease in cell number after 48 h. Cellular death was attributable to apoptosis based on cellular blebbing, caspase-3 activation, vital dye and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining, and poly (ADP-ribose) polymerase (PARP) cleavage. Transporter knockdown also markedly increased activities of caspases-2 and -9, marginally enhanced caspase-8 activity, and dramatically increased ASCT1 mRNA levels, presumably as a futile compensatory response. Apoptosis elicited via transporter silencing was not attributable to the double-stranded RNA-dependent protein kinase R (PKR) pathway. For comparison, glutamine deprivation also caused apoptotic cell death but with slower temporal kinetics, stimulated caspases-2 and -3 but not caspases-8 or -9 activities, and led to considerable PARP cleavage. Thus ASCT2 suppression exerts proapoptotic effects transcending those of glutamine starvation alone. We conclude that ATB(0)/ASCT2 expression is necessary for SK-Hep cell growth and viability and suggest that it be further explored as a selective target for human hepatocellular carcinoma. PMID- 14563675 TI - Gender differences in small intestinal endothelial function: inhibitory role of androgens. AB - Although gender differences exist in cardiovascular endothelial function, it remains unclear whether such differences are also seen in small intestinal endothelial function. To determine this, untreated male, age-matched proestrus female, castrated male, and 17beta-estradiol (E2)-treated noncastrated male rats were studied. Dose response curves to ACh and nitroglycerin (NTG) were determined by measuring changes in perfusion pressure by using an isolated small intestinal perfusion model. Endothelium-derived nitric oxide (NO) production/release was indirectly determined by the ability of intact endothelium to suppress serotonin (10(-5) M)-induced perfusion pressure changes. Intestinal tissue levels of NO were also measured. Moreover, plasma levels of androgen and E2 were determined and correlated with ACh (10(-8) M)-induced perfusion pressure reductions. ACh induced intestinal perfusion pressure reductions in proestrus females, castrated males, and E2-treated noncastrated males were significantly higher than in untreated males. NTG-induced perfusion pressure reductions were not significantly different among groups. Perfusion pressures after administration of serotonin (10(-5) M) and intestinal tissue levels of NO in proestrus females, castrated males, and E2-treated noncastrated males were also significantly higher than in untreated males. Plasma androgen levels in proestrus females, castrated males, and in E2-treated noncastrated males were significantly lower compared with untreated males. There was a positive correlation between plasma androgen and ACh reduced perfusion pressure; however, E2 levels did not show a similar relationship. Thus androgens appear to play an inhibitory role in small intestinal endothelial function. These properties in male vessels can be modulated by decreasing the level of circulating androgens or by E2 treatment. PMID- 14563676 TI - Tomoregulin-1 (TMEFF1) inhibits nodal signaling through direct binding to the nodal coreceptor Cripto. AB - Transforming growth factor beta (TGF-beta) signals regulate multiple processes during development and in adult. We recently showed that tomoregulin-1 (TMEFF1), a transmembrane protein, selectively inhibits nodal but not activin in early Xenopus embryos. Here we report that TMEFF1 binds to the nodal coreceptor Cripto, but does not associate with either nodal or the type I ALK (activin receptor-like kinase) 4 receptor in coimmunoprecipitation assays. The inhibition of the nodal signaling by TMEFF1 in Xenopus ectodermal explants is rescued with wild-type but not mutant forms of Cripto. Furthermore, we show that the Cripto-FRL1-Cryptic (CFC) domain in Cripto, which is essential for its binding to ALK4, is also important for its interaction with TMEFF1. Our results demonstrate for the first time that nodal signaling can be regulated by a novel mechanism of blocking the Cripto coreceptor. PMID- 14563677 TI - Regulator of sex-limitation (Rsl) encodes a pair of KRAB zinc-finger genes that control sexually dimorphic liver gene expression. AB - Sexually dimorphic expression of a broad array of liver proteins involved in reproduction and xenobiotic metabolism is induced at puberty by sex-specific growth hormone patterns. An additional control of sex-dependent gene expression is conferred by Regulator of sex-limitation (Rsl) alleles. In variant rsl mice, females inappropriately express the male Sex-limited protein, Slp. We recently showed that a panel of male-specific liver genes is repressed by Rsl, accentuating sex differences in a hormone-independent manner. Here we map rsl to a region on Chromosome 13 comprised exclusively of KRAB (Kruppel-associated box) zinc-finger protein (ZFP) genes. Among eight Rsl candidate (Rslcan) genes within the critical genetic interval, the recent duplicates Rslcan-4 and Rslcan-9 both harbor mutations in rsl mice (partial deletion and splice-site inactivation, respectively). Transgenesis with bacterial artificial chromosome (BAC) clones encompassing Rslcan-4 restores male-specific MUP (major urinary protein) expression to rsl mice, whereas a BAC containing Rslcan-9 rescues sex-specific expression of Slp and cytochrome P450 Cyp2d9. Thus, the Rslcan-4 and Rslcan-9 paralogs partitioned regulation of their target genes during evolution. This demonstrates the first biological role for a set of KRAB zinc-finger repressor proteins and reveals the molecular basis of a gene-silencing pathway critical for sexual dimorphism. PMID- 14563678 TI - Ceramides modulate programmed cell death in plants. AB - The balance between the bioactive sphingolipid ceramide and its phosphorylated derivative has been proposed to modulate the amount of programmed cell death (PCD) in eukaryotes. We characterized the first ceramide kinase (CERK) mutant in any organism. The Arabidopsis CERK mutant, called accelerated cell death 5, accumulates CERK substrates and shows enhanced disease symptoms during pathogen attack and apoptotic-like cell death dependent on defense signaling late in development. ACD5 protein shows high specificity for ceramides in vitro. Strikingly, C2 ceramide induces, whereas its phosphorylated derivative partially blocks, plant PCD, supporting a role for ceramide phosphorylation in modulating cell death in plants. PMID- 14563679 TI - Transcriptional activation via sequential histone H2B ubiquitylation and deubiquitylation, mediated by SAGA-associated Ubp8. AB - Gene activation and repression regulated by acetylation and deacetylation represent a paradigm for the function of histone modifications. We provide evidence that, in contrast, histone H2B monoubiquitylation and its deubiquitylation are both involved in gene activation. Substitution of the H2B ubiquitylation site at Lys 123 (K123) lowered transcription of certain genes regulated by the acetylation complex SAGA. Gene-associated H2B ubiquitylation was transient, increasing early during activation, and then decreasing coincident with significant RNA accumulation. We show that Ubp8, a component of the SAGA acetylation complex, is required for SAGA-mediated deubiquitylation of histone H2B in vitro. Loss of Ubp8 in vivo increased both gene-associated and overall cellular levels of ubiquitylated H2B. Deletion of Ubp8 lowered transcription of SAGA-regulated genes, and the severity of this defect was exacerbated by codeletion of the Gcn5 acetyltransferase within SAGA. In addition, disruption of either ubiquitylation or Ubp8-mediated deubiquitylation of H2B resulted in altered levels of gene-associated H3 Lys 4 methylation and Lys 36 methylation, which have both been linked to transcription. These results suggest that the histone H2B ubiquitylation state is dynamic during transcription, and that the sequence of histone modifications helps to control transcription. PMID- 14563680 TI - Meiotic double-strand breaks at the interface of chromosome movement, chromosome remodeling, and reductional division. AB - Chromosomal processes related to formation and function of meiotic chiasmata have been analyzed in Sordaria macrospora. Double-strand breaks (DSBs), programmed or gamma-rays-induced, are found to promote four major events beyond recombination and accompanying synaptonemal complex formation: (1) juxtaposition of homologs from long-distance interactions to close presynaptic coalignment at midleptotene; (2) structural destabilization of chromosomes at leptotene/zygotene, including sister axis separation and fracturing, as revealed in a mutant altered in the conserved, axis-associated cohesin-related protein Spo76/Pds5p; (3) exit from the bouquet stage, with accompanying global chromosome movements, at zygotene/pachytene (bouquet stage exit is further found to be a cell-wide regulatory transition and DSB transesterase Spo11p is suggested to have a new noncatalytic role in this transition); (4) normal occurrence of both meiotic divisions, including normal sister separation. Functional interactions between DSBs and the spo76-1 mutation suggest that Spo76/Pds5p opposes local destabilization of axes at developing chiasma sites and raise the possibility of a regulatory mechanism that directly monitors the presence of chiasmata at metaphase I. Local chromosome remodeling at DSB sites appears to trigger an entire cascade of chromosome movements, morphogenetic changes, and regulatory effects that are superimposed upon a foundation of DSB-independent processes. PMID- 14563681 TI - N-Glycosylation-dependent block is a novel mechanism for drug-induced cardiac arrhythmia. AB - Voltage-gated potassium channels formed with the cardiac subunit HERG and a polymorphic variant of MinK-related peptide 1 (MiRP1) exhibit increased susceptibility to the antibiotic sulfamethoxazole (SMX) compared with channels formed with wild-type (WT) subunits. Here the molecular bases for SMX high affinity block are investigated. The polymorphism causes a benign T to A amino acid mutation at position 8 (T8A) that destroys an N-glycosylation site of MiRP1. In vitro disruption of glycosylation by mutagenesis or in vivo by treatment with neuraminidase is associated with increased susceptibility to SMX and to other elementary agents such as divalent cations. Defective glycosylation does not affect the ability of T8A to form stable complexes with HERG, but rather it increases drug susceptibility through structural modifications in the channel complex. We conclude that N-glycosylation may play a key role in the etiology of life-threatening arrhythmia. PMID- 14563682 TI - Resveratrol induces growth inhibition and apoptosis in metastatic breast cancer cells via de novo ceramide signaling. AB - Resveratrol (3,4',5-trans-trihydroxystilbene), a phytoalexin present in grapes and red wine, is emerging as a natural compound with potential anticancer properties. Here we show that resveratrol can induce growth inhibition and apoptosis in MDA-MB-231, a highly invasive and metastatic breast cancer cell line, in concomitance with a dramatic endogenous increase of growth inhibitory/proapoptotic ceramide. We found that accumulation of ceramide derives from both de novo ceramide synthesis and sphingomyelin hydrolysis. More specifically we demonstrated that ceramide accumulation induced by resveratrol can be traced to the activation of serine palmitoyltransferase (SPT), the key enzyme of de novo ceramide biosynthetic pathway, and neutral sphingomyelinase (nSMase), a main enzyme involved in the sphingomyelin/ceramide pathway. However, by using specific inhibitors of SPT, myriocin and L-cycloserine, and nSMase, gluthatione and manumycin, we found that only the SPT inhibitors could counteract the biological effects induced by resveratrol. Thus, resveratrol seems to exert its growth inhibitory/apoptotic effect on the metastatic breast cancer cell line MDA-MB-231 by activating the de novo ceramide synthesis pathway. PMID- 14563683 TI - Discovery of a potent nanoparticle P-selectin antagonist with anti-inflammatory effects in allergic airway disease. AB - The severity of allergic asthma is dependent, in part, on the intensity of peribronchial inflammation. P-selectin is known to play a role in the development of allergen-induced peribronchial inflammation and airway hyperreactivity. Selective inhibitors of P-selectin-mediated leukocyte endothelial-cell interactions may therefore attenuate the inflammatory processes associated with allergic airway disease. Novel P-selectin inhibitors were created using a polyvalent polymer nanoparticle capable of displaying multiple synthetic, low molecular weight ligands. By assembling a particle that presents an array of groups, which as monomers interact with only low affinity, we created a construct that binds extremely efficiently to P-selectin. The ligands acted as mimetics of the key binding elements responsible for the high-avidity adhesion of P-selectin to the physiologic ligand, PSGL-1. The inhibitors were initially evaluated using an in vitro shear assay system in which interactions between circulating cells and P-selectin-coated capillary tubes were measured. The nanoparticles were shown to preferentially bind to selectins expressed on activated endothelial cells. We subsequently demonstrated that nanoparticles displaying P-selectin blocking arrays were functionally active in vivo, significantly reducing allergen-induced airway hyperreactivity and peribronchial eosinophilic inflammation in a murine model of asthma. PMID- 14563684 TI - Transcription of stem cell factor (SCF) is potentiated by glucocorticoids and interleukin-1beta through concerted regulation of a GRE-like and an NF-kappaB response element. AB - Expression of stem cell factor SCF, a major mast cell growth factor, is potentiated shortly after co-treatment with interleukin (IL)-1beta and glucocorticoids. SCF promoter contains a GRE-like sequence and a putative kappaB site. We assessed the mechanisms of the regulation of SCF transcription in human lung fibroblasts in culture. Chromatin immunoprecipitation showed that co treatment with IL-1beta and the glucocorticoid budesonide increased the SCF promoter occupancy by NF-kappaB and GR, as compared with IL-1beta and budesonide alone. In reporter gene assays, IL-1beta time-dependently increased the promoter activity, which was abolished by either pre-treatment with the MAP kinase inhibitors PD98059 (MEK) and SB203580 (p38), pre-treatment with the NF-kappaB inhibitor PDTC, or deletion of the kappaB site. Budesonide time-dependently decreased the promoter activity, an effect requiring the GRE-like element. Co treatment with IL-1beta and budesonide potentiated the promoter activity at 30 min, an effect blocked by PD98059 and SB203580, PDTC, or deletion of the kappaB or GRE-like element. In conclusion, the GRE-like sequence mediating the repression of SCF expression, thus acting as a negative-responsive element, is turned into a positive element in an NF-kappaB site-dependent manner, indicating a concerted action of these two regulatory elements in the potentiation of SCF gene expression. PMID- 14563685 TI - Ischemia-reperfusion selectively impairs nitric oxide-mediated dilation in coronary arterioles: counteracting role of arginase. AB - A reduction in L-arginine availability has been implicated in the impairment of endothelium-dependent nitric oxide (NO)-mediated vasodilation by ischemia reperfusion (I/R). However, the mechanisms contributing to dysregulation of the L arginine pool remain unknown. Because endothelial cells can metabolize L-arginine via two major enzymes, that is, NO synthase (NOS) and arginase, we hypothesized that up-regulation of arginase during I/R reduces L-arginine availability to NOS and thus impairs NO-mediated vasodilation. To test this hypothesis, a local I/R was produced in the porcine heart by occlusion of a small branch of left anterior descending artery for 30 min, followed by reperfusion for 90 min. Arterioles (60 110 microm) isolated from non-ischemic and ischemic regions of subepicardium were cannulated and pressurized without flow for in vitro study. Vessels from both regions developed similar levels of basal tone. Although the dilation of I/R vessels to endothelium-independent agonist sodium nitroprusside was not altered, the endothelium-dependent NO-mediated dilations to adenosine and serotonin were attenuated. I/R not only inhibited arteriolar production of NO but also increased arteriolar arginase activity. Arginase inhibitor alpha-difluoromethylornithine enhanced NO production/dilation in normal vessels and also restored the NO mediated function in I/R vessels. Treating I/R vessels with L-arginine also restored vasodilations. Immunohistochemical data revealed that I/R up-regulated arginase but down-regulated NOS expression in the arteriolar endothelium. Pretreating the animals with protein synthesis inhibitor cycloheximide prevented I/R-induced arginase up-regulation and also preserved NO-mediated vascular function. These results suggest that one mechanism by which I/R inhibits NO mediated arteriolar dilation is through increased arginase activity, which limits the availability of L-arginine to NOS for NO production. In addition, the inability of arginase blockade or L-arginine supplementation to completely restore vasodilatory function may be attributable to the down-regulation of endothelial NOS expression. PMID- 14563686 TI - MMP-2 and MMP-9 synergize in promoting choroidal neovascularization. AB - Matrix metalloproteinase 2 (MMP-2) and MMP-9 are increased in human choroidal neovascularization (CNV) occurring during the exudative most aggressive form of age-related macular degeneration (AMD), but their precise role and potential interactions remain unclear. To address the question of MMP-2 and MMP-9 functions, mice deficient in the expression of MMP-2 (MMP-2 KO), MMP-9 (MMP-9 KO), and both MMP-2 and MMP-9 (MMP-2,9 KO) with their corresponding wild-type mice (WT) underwent CNV induction by laser-induced rupture of the Bruch's membrane. Both the incidence and the severity of CNV were strongly attenuated in double deficient compared with single gene deficient mice or corresponding WT controls. The reduced neovascularization was accompanied by fibrinogen/fibrin accumulation. Furthermore, overexpression of the endogenous MMP inhibitors TIMP-1 or TIMP-2 (delivered by adenoviral vectors) in WT mice or daily injection of a synthetic and gelatinase selective MMP inhibitor (Ro 26-2853) significantly decreased the pathological reaction. These findings suggest that MMP-2 and MMP-9 may cooperate in the development of AMD and that their selective inhibition represents an alternative strategy for the treatment of choroidal neovascularization. PMID- 14563687 TI - An atomic detail model for the human ATP binding cassette transporter P glycoprotein derived from disulfide cross-linking and homology modeling. AB - The multidrug resistance P-glycoprotein mediates the extrusion of chemotherapeutic drugs from cancer cells. Characterization of the drug binding and ATPase activities of the protein have made it the paradigm ATP binding cassette (ABC) transporter. P-glycoprotein has been imaged at low resolution by electron cryo-microscopy and extensively analyzed by disulphide cross-linking, but a high resolution structure solved ab initio remains elusive. Homology models of P-glycoprotein were generated using the structure of a related prokaryotic ABC transporter, the lipid A transporter MsbA, as a template together with structural data describing the dimer interface of the nucleotide binding domains (NBDs). The first model, which maintained the NBD:transmembrane domain (TMD) interface of MsbA, did not satisfy previously published cross-linking data. This suggests that either P-glycoprotein has a very different structure from MsbA or that the published E. coli MsbA structure does not reflect a physiological state. To distinguish these alternatives, we mapped the interface between the two TMDs of P glycoprotein experimentally by chemical cross-linking of introduced triple cysteine residues. Based on these data, a plausible atomic model of P glycoprotein could be generated using the MsbA template, if the TMDs of MsbA are reoriented with respect to the NBDs. This model will be important for understanding the mechanism of P-glycoprotein and other ABC transporters. PMID- 14563688 TI - Myosin heavy chain expression in renal afferent and efferent arterioles: relationship to contractile kinetics and function. AB - The physiological role of smooth muscle myosin heavy chain (MHC) isoform diversity is poorly understood. The expression of MHC-B, which contains an insert at the ATP binding pocket, has been linked to enhanced contractile kinetics. We recently reported that the renal afferent arteriole exhibits an unusually rapid myogenic response and that its kinetic features allow this vessel to modulate tone in response to alterations in systolic blood pressure. In the present study, we examined MHC expression patterns in renal afferent and efferent arterioles. These two vessels regulate glomerular inflow and outflow resistances and control the pressure within the intervening glomerular capillaries (PGC). Whereas the afferent arteriole must respond rapidly to increases in blood pressure, the efferent arteriole plays a distinctly different role, maintaining a tonic elevation in outflow resistance to preserve function when renal perfusion is compromised. Using RT-PCR, Western analysis, and immunofluorescence imaging of intact isolated arterioles, we found that the afferent arteriole predominantly expresses the MHC-B isoform, whereas the efferent arteriole expresses only the slower-cycling MHC-A isoform. We examined the kinetics of angiotensin II- and norepinephrine-induced vasoconstriction and found that the afferent arteriole responds approximately 3-fold faster than the efferent arteriole. Our findings thus point to the renal microcirculation as a unique and important example of smooth muscle adaptation in regard to MHC isoform expression and physiological function. PMID- 14563689 TI - Rapid induction of autoantibodies against Nogo-A and MOG in the absence of an encephalitogenic T cell response: implication for immunotherapeutic approaches in neurological diseases. AB - Vaccinations against various antigens of the central nervous system (CNS) are gaining increasing interest as a therapeutic approach in a variety of neurological diseases such as spinal cord injury, ischemic stroke, Alzheimer disease, or spongiform encephalopathy. In the present work, the time window after spinal cord injury allowing potentially therapeutic antibody to penetrate the damaged blood-brain barrier (BBB) was measured by intravenous injection of a monoclonal anti-Nogo-A antibody. Although an influx of Nogo antibodies at the lesion site was detectable up to 2 wk after injury, a significant decrease in BBB permeability was noticed within the first week. Clearly, therefore, a vaccination protocol with a rapid antibody response is required for acute therapeutic interventions after CNS trauma. We designed a conjugate vaccine paradigm with particular focus on the safety and the kinetics of the antibody response. As antigen targets, we used Nogo-A and the strongly encephalitogenic myelin oligodendrocyte glycoprotein (MOG). Intrasplenic autoimmunization of rats with a Nogo-A-specific region fused to the Tetanus toxin C-fragment (TTC) resulted in a fast IgM response against Nogo-A. A specific switch to IgG was observed as soon as 4-7 days after intrasplenic immunization in TTC-primed animals. In spite of the induction of a specific IgG response after intrasplenic immunization, no signs of experimental autoimmune disease (EAE) or inflammatory infiltrates on histological examinations were observable. In contrast to subcutaneous immunization with MOG, in vitro cytokine secretion assays (IL-2, IL-10, and IFN gamma) did not reveal activation of MOG-specific T cells after intrasplenic immunization. Our findings have critical implications for future strategies in the development of safe and efficient therapeutic vaccines for neurological diseases. PMID- 14563690 TI - Inducible cyclooxygenase-derived 15-deoxy(Delta)12-14PGJ2 brings about acute inflammatory resolution in rat pleurisy by inducing neutrophil and macrophage apoptosis. AB - Failure of acute inflammation to resolve leads to persistence of the inflammatory response and may contribute to the development of chronic inflammation. Thus, an understanding of inflammatory resolution will provide insight into the etiology of chronic inflammation. In an acute pleurisy, polymorphonuclear leukocytes (PMNs) were found to predominate at the onset of the lesion but decreased in number by undergoing apoptosis, the principal mechanism by which PMNs died in this model. PMNs were progressively replaced by monocytes, which differentiated into macrophages. As with PMNs, macrophages also underwent programmed cell death leading to an abatement of the inflammatory response and eventual resolution. It was found that apoptosis of both these inflammatory cell types was mediated by pro-resolving cyclooxygenase 2-derived 15deoxyDelta12-14PGJ2, which is uniquely expressed during active resolution. Although PMN programmed cell death is well understood, the observation that macrophages apoptose during resolution of acute inflammation is less well described. These results provide insight into the mechanisms that switch off acute inflammation and prevent complications of wound healing and potentially the development of immune-mediated chronic inflammation. PMID- 14563691 TI - Alzheimer's disease skin fibroblasts selectively express a bradykinin signaling pathway mediating tau protein Ser phosphorylation. AB - Increased Ser phosphorylation of tau microtubule-associated protein in the brain is an early feature of Alzheimer's disease (AD) that precedes progression of the disease to frank neuronal disruption. We demonstrate that bradykinin (BK) B2 receptor activation leads to selective Ser phosphorylation of tau in skin fibroblasts from persons who have or will develop AD due to Presenilin 1 mutations or Trisomy 21, but not in skin fibroblasts from normal individuals at any age. The increased signal transduction in AD fibroblasts that culminates in tau Ser phosphorylation reflects modification of the G protein-coupled BK B2 receptors themselves. Both the BK B2 receptor modification and BK-mediated tau Ser phosphorylation are dependent on activation of protein kinase C and can be detected in fibroblasts from persons with Trisomy 21 two decades before the characteristic onset of AD. This dysregulated signaling cascade in AD may thus be expressed throughout life as an aberrant pathway in peripheral tissues more accessible than brain for molecular analysis. The sites of greatest BK B2 receptor expression in brain overlap with those areas displaying the earliest pathology in the course of AD, suggesting that BK receptor pathway dysfunction may be a molecular signature yielding information about the pathogenesis of AD. PMID- 14563692 TI - Complement C3a receptors in the pituitary gland: a novel pathway by which an innate immune molecule releases hormones involved in the control of inflammation. AB - Two-way communication exists between the endocrine and immune systems using molecules such as hormones and cytokines. Here we describe a new pathway by which C3a, a complement-derived cytokine, stimulates anterior pituitary hormone release and activates the hypothalamic-pituitary-adrenal axis, a reflex central to the stress response and to the control of inflammation. We show that C3a receptors are expressed in pituitary hormone secreting and non-hormone secreting (folliculostellate) cells and that both C3a and C3adesArg (a non-inflammatory metabolite) stimulate pituitary cell cultures to release prolactin, growth hormone, and adrenocorticotropin. Serum levels of these hormones, together with adrenal corticosterone, increase dose dependently with recombinant C3a and C3adesArg administration in vivo. Pertussis toxin blocks the response to C3a but not C3adesArg, which indicates the presence of two receptors, only one of which is coupled to Galphai-proteins. We propose that the complement innate immune molecules (cytokines) modulate tissue-specific and systemic inflammatory responses through communication with the endocrine pituitary gland. PMID- 14563693 TI - Cell-demanded release of VEGF from synthetic, biointeractive cell ingrowth matrices for vascularized tissue growth. AB - Local, controlled induction of angiogenesis remains a challenge that limits tissue engineering approaches to replace or restore diseased tissues. We present a new class of bioactive synthetic hydrogel matrices based on poly(ethylene glycol) (PEG) and synthetic peptides that exploits the activity of vascular endothelial growth factor (VEGF) alongside the base matrix functionality for cellular ingrowth, that is, induction of cell adhesion by pendant RGD-containing peptides and provision of cell-mediated remodeling by cross-linking matrix metalloproteinase substrate peptides. By using a Michael-type addition reaction, we incorporated variants of VEGF121 and VEGF165 covalently within the matrix, available for cells as they invade and locally remodel the material. The functionality of the matrix-conjugated VEGF was preserved and was critical for in vitro endothelial cell survival and migration within the matrix environment. Consistent with a scheme of locally restricted availability of VEGF, grafting of these VEGF-modified hydrogel matrices atop the chick chorioallontoic membrane evoked strong new blood vessel formation precisely at the area of graft-membrane contact. When implanted subcutaneously in rats, these VEGF-containing matrices were completely remodeled into native, vascularized tissue. This type of synthetic, biointeractive matrix with integrated angiogenic growth factor activity, presented and released only upon local cellular demand, could become highly useful in a number of clinical healing applications of local therapeutic angiogenesis. PMID- 14563694 TI - Urocortin protects cardiac myocytes from ischemia/reperfusion injury by attenuating calcium-insensitive phospholipase A2 gene expression. AB - We have used Affymetrix gene chip technology to look for changes in gene expression caused by a 24 h exposure of rat primary neonatal cardiac myocytes to the cardioprotective agent urocortin. We observed a 2.5-fold down-regulation at both the mRNA and protein levels of a specific calcium-insensitive phospholipase A2 enzyme. Levels of lysophosphatidylcholine, a toxic metabolite of phospholipase A2, were lowered by 30% in myocytes treated with urocortin for 24 h and by 50% with the irreversible iPLA2 inhibitor bromoenol lactone compared with controls. Both 4 h ischemia and ischemia followed by 24 h reperfusion caused a significant increase in lysophosphatidylcholine concentration compared with controls. When these myocytes were pretreated with urocortin, the ischemia-induced increase in lysophosphatidylcholine concentration was significantly lowered. Moreover, co incubation of cardiac myocytes with urocortin, or the specific phospholipase A2 inhibitor bromoenol lactone, reduces the cytotoxicity produced by lysophosphatidylcholine or ischemia/reperfusion. Similarly, in the intact heart ex vivo we found that cardiac damage measured by infarct size was significantly increased when lysophoshatidylcholine was applied during ischemia, compared with ischemia alone, and that pre-treatment with both urocortin and bromoenol lactone reversed the increase in infarct size. This, to our knowledge, is the first study linking the cardioprotective effect of urocortin to a decrease in a specific enzyme protein and a subsequent decrease in the concentration of its cardiotoxic metabolite. PMID- 14563695 TI - Genetic predisposition to the cytotoxicity of arsenic: the role of DNA damage and ATM. AB - Arsenic is a pervasive cytotoxin and carcinogen in the environment. Although its mode of action has yet to be fully elucidated, oxidative DNA damage has been suggested. A series of DNA repair-defective human and hamster cell lines associated with sensitivity to oxidative agents were examined for their response to arsenic-induced cytotoxicity. Only the Ataxia telangiectasia (AT) cells displayed a marked hypersensitive response (greater than twofold). The protective role of the ATM protein was confirmed by the normal response to arsenic displayed by AT cells expressing wild-type ATM. Although the ATM protein plays a pivotal role in response to DNA double-strand breakage, none of the other cell lines with defects in double-strand break repair displayed a similar hypersensitivity. Further examination indicated that concentrations of sodium arsenite as high as 1 mg/l do not generate significant levels of double-strand breaks. Our data suggest that the ATM protein functions in an important but different capacity in the cellular response to arsenic toxicity than it does in response to agents that generate double-strand breaks, such as ionizing radiation. Furthermore, the lack of hypersensitivity to arsenic displayed by the other cell lines calls into question the hypothesis that DNA damage is a significant factor in arsenic cytotoxicity. PMID- 14563696 TI - Corticotropin-releasing hormone and arginine vasopressin gene transcription in the hypothalamic paraventricular nucleus of unstressed rats: daily rhythms and their interactions with corticosterone. AB - To drive the daily ACTH secretory rhythm from anterior pituitary corticotropes signals from the circadian clock schedule CRH and, to lesser extent, arginine vasopressin (AVP) release from neuroendocrine terminals. In turn, releasable pools of CRH and AVP in neuroendocrine terminals are sustained by synthetic mechanisms in the medial parvicellular paraventricular nucleus, a critical component of which involves transcribing primary (heteronuclear (hn)) RNA transcripts from their cognate genes. To determine the fundamental daily patterns of ACTH secretagogue gene transcription in unstressed rats, we measured CRH and AVP hnRNA levels at 1- to 4-h intervals throughout the day using in situ hybridization. Crh gene transcription is readily detectable throughout the day, and shows a pronounced rhythm that is temporally correlated with CRH mRNA levels, but is uncoupled from ACTH release. However, avp gene transcription is barely detectable and shows no discernable rhythm. We then performed similar experiments in adrenalectomized rats with or without corticosterone replacement. Corticosterone-dependent mechanisms regulate CRH hnRNA levels at the nadir and peak as well as the onset of nocturnal crh gene transcription. A prominent rhythm of avp hnRNA seen in adrenalectomized animals was dampened by corticosterone. This study shows, first, CRH synthesis in intact animals is maintained by a nocturnal episode of crh gene transcription, parameters of which are modulated by corticosterone-dependent mechanisms; second, circulating corticosterone is sufficient to completely inhibit a daily rhythm of avp gene transcription present in adrenalectomized rats; third, the neural systems that activate crh gene transcription can be uncoupled from those driving ACTH release. PMID- 14563697 TI - Androgen-sensitive changes in regulation of restraint-induced adrenocorticotropin secretion between early and late puberty in male rats. AB - Regulation of ACTH secretion changes between early (40 d) and late (60 d) puberty in male rats. We tested whether this occurs because of activating effects of testosterone on the brain. We measured testosterone and ACTH responses to repeated restraint in adrenalectomized, corticosterone-replaced rats entering and leaving puberty with or without treatment with flutamide, a nonsteroidal androgen receptor antagonist. Flutamide increased testosterone. ACTH responses were high and suppressed by flutamide at 40 d. At 60 d, ACTH responses were low and increased by flutamide. On d 4, basal arginine vasopressin (AVP) mRNA was increased by restraint, but not age, in the medial parvicellular paraventricular nucleus (mpPVN) and medial amygdala and increased with age in the bed nucleus of the stria terminalis. We counted numbers of AVP-immunoreactive (AVP-ir) and corticotropin-releasing factor (CRF)-ir neurons. In medial amygdala, there was no change in AVP+ cells. With restraint, CRF+ cells in the central nucleus decreased at 40 d and increased at 60 d. Flutamide did not affect the response at 40 d but blocked restraint-induced increases at 60 d. After restraint, the bed nucleus of the stria terminalis AVP-ir correlated negatively with mpPVN CRF-ir at 40 d and with mpPVN AVP-ir at 60 d. In PVN, there were no effects on CRF+ cells. However, AVP+ cells increased only with restraint plus flutamide at 40 d and tended to increase with restraint and decrease with restraint plus flutamide at 60 d. We conclude that during puberty testosterone induces marked changes in regulation of neuropeptides in pathways known to determine autonomic, neuroendocrine, and behavioral responses to chronic stress. PMID- 14563698 TI - Functional modification of pituitary somatotropes in the aromatase knockout mouse and the effect of estrogen replacement. AB - Available data on the influence of estradiol (E(2)) on GH levels remains controversial. A factor contributing to this uncertainty is a lack of knowledge of both E(2) action on somatotropes as well as the molecular mechanisms involved. In this study we investigated gene expression implicated in GH secretion in somatotropes derived from female aromatase knockout (ArKO) mice. In these mice E(2) production is blocked due to disruption of the Cyp19 gene encoding aromatase, the enzyme responsible for estrogen biosynthesis. The effect of E(2) replacement was also studied by in vivo treatment of mice with E(2) for 3 wk. It was demonstrated that somatotropes from ArKO mice had a low expression of GH, GH secretagogue receptor, GHRH receptor (GHRH-R), and pituitary-specific transcription factor (Pit-1). On the other hand, the somatotropes exhibited elevated expression of somatostatin receptors (sst1-5). Overall, these effects resulted in a reduction in GH secretion. E(2) replacement increased GHRH-R, Pit 1, and GH mRNA levels to 185%, 193%, and 157% and reduced the levels of sst1, sst2, sst4, and sst5 mRNA expression in ArKO mice, respectively. E(2) replacement did not affect the levels of pituitary estrogen (alpha and beta) and androgen receptor mRNA expression. It is concluded that the expression of important genes involved in GH synthesis in somatotropes of the female ArKO mouse are functionally down-regulated, and such a down-regulation is reversed to normal levels by E(2) replacement. The levels of GH secretagogue receptor, GHRH-R, and Pit-1 mRNA expression were also reduced, and sst1 and sst3 mRNA expression enhanced in aging ArKO and wild-type mice, resulting in a decrease in GH mRNA expression. It is suggested that aging is another important impact factor for the pituitary expression and regulation of GH mRNA in female mice. PMID- 14563699 TI - Nitric oxide regulates receptor activator of nuclear factor-kappaB ligand and osteoprotegerin expression in bone marrow stromal cells. AB - Bone remodeling reflects an equilibrium between bone resorption and formation. The local expression of receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) in bone determines the entry of monoblastic precursors into the osteoclast lineage and subsequent bone resorption. Nitric oxide (NO) inhibits osteoclastic bone resorption in vitro and regulates bone remodeling in vivo. An interaction of NO with RANKL and OPG has not been studied. Here, we show that treatment of ST-2 murine stromal cells with the NO donor sodium nitroprusside (100 microm) for 24 h inhibited 1,25 dihydroxyvitamin D(3) induced RANKL mRNA to less than 33 +/- 7% of control level, whereas OPG mRNA increased to 204 +/- 19% of control. NOR-4 replicated these NO effects. The effects of NO were dose dependent and associated with changes in protein levels: RANKL protein decreased and OPG protein increased after treatment with NO. PTH induced RANKL expression in primary stromal cells was inhibited by sodium nitroprusside, indicating that the NO effect did not require vitamin D. NO donor did not change the stability of RANKL or OPG mRNAs, suggesting that NO affected transcription. Finally, cGMP, which can function as a second messenger for NO, did not reproduce the NO effect, nor did inhibition of endogenous guanylate cyclase prevent the NO effect on these osteoactive genes. The effect of NO to decrease the RANKL/OPG equilibrium should lead to decreased recruitment of osteoclasts and positive bone formation. Thus, drugs and conditions that cause local increase in NO formation in bone may have positive effects on bone remodeling. PMID- 14563700 TI - Phosphatidylinositol 3-kinase activation is required for sulfonylurea stimulation of glucose transport in rat skeletal muscle. AB - Sulfonylureas are drugs widely used in the treatment of patients with type 2 diabetes mellitus. In addition to their pancreatic effect of stimulating insulin secretion, many studies suggest that sulfonylureas also have extrapancreatic actions. We have previously reported that gliclazide, a second-generation sulfonylurea, stimulates the glucose uptake by rat hindquarter skeletal muscle directly and immediately by promoting the translocation of glucose transporter 4 to the plasma membrane. The aim of our study was to approach the gliclazide intracellular signaling pathway. For this purpose, we incubated clamped and isolated soleus muscle from rat with gliclazide. The following results were obtained: 1) gliclazide stimulates insulin receptor substrate (IRS)-1 phosphatidylinositol 3 (PI3)-kinase-associated activity, and this activity is necessary for gliclazide-stimulated glucose transport; 2) gliclazide treatment produces a gradual translocation of the diacylglycerol (DAG)-dependent isoforms protein kinase C (PKC) alpha, theta, and epsilon from cytosolic to membrane fraction that is dependent on PI3-kinase and phospholipase C (PLC)-gamma activation; and 3) PKC and PLC-gamma activation is necessary for gliclazide stimulated glucose transport. We propose a hypothetical signaling pathway by which gliclazide could stimulate IRS-1 that would allow its association with PI3 kinase, promoting its activation. PI3-kinase products could induce PLC-gamma activation, whose hydrolytic activity could activate the DAG-dependent isoforms PKC alpha, theta, and epsilon. PMID- 14563701 TI - Involvement of cholesterol-rich lipid rafts in interleukin-6-induced neuroendocrine differentiation of LNCaP prostate cancer cells. AB - IL-6 is an inflammatory cytokine that has been linked to aggressive prostate cancer (PCa). Previous studies have demonstrated that IL-6 can enhance the differentiation of PCa cells toward a neuroendocrine (NE) phenotype, a possible indicator of hormone-refractory disease. In this report, we present evidence that the mechanism of IL-6-stimulated NE differentiation employs a detergent-resistant (lipid raft) membrane compartment for signal transduction in LNCaP PCa cells. Signal transducer and activator of transcription (STAT)3, a mediator of IL-6 signaling, was rapidly phosphorylated and translocated to the nucleus in LNCaP cells treated with IL-6. Both processes were inhibited by filipin, a cholesterol binding compound that disrupts plasma membrane lipid rafts. Isolation of Triton X 100-insoluble raft fractions from LNCaP cells by discontinuous sucrose gradient centrifugation demonstrated that the 80-kDa IL-6 receptor localized almost exclusively to the raft compartment. Although STAT3 was located predominantly in the Triton X-100-soluble subcellular fraction in exponentially growing cells, abundant phosphorylated STAT3 was detected in the raft fraction after stimulation with IL-6. Increases in expression of the NE marker, neuron-specific enolase, and neuron-specific enolase promoter activity after IL-6 treatment were reduced after membrane rafts were disrupted by filipin treatment. LNCaP cells expressed the raft-resident proteins flotillin-2 and G(ialpha2), but notably not caveolins, the predominant structural protein present in caveolar membrane rafts in many tissues and tumor cells. These results are the first to define a role for lipid raft membrane microdomains in signal transduction mechanisms capable of promoting the NE phenotype in PCa cells, and they demonstrate that the raft compartment is capable of mediating such signals in the absence of caveolins. Our results also suggest a mechanistic role for membrane cholesterol in cell signaling events relevant to PCa progression. PMID- 14563702 TI - The role of cyclic-ADP-ribose-signaling pathway in oxytocin-induced Ca2+ transients in human myometrium cells. AB - Human myometrial contraction plays a fundamental role in labor. Dysfunction of uterine contraction is an important cause of labor progression failure. Although the mechanisms controlling uterine contraction are not completely understood, intracellular Ca2+ mobilization plays an important role during uterine contraction. Several mechanisms of intracellular Ca2+ mobilization are present in smooth muscle, but in the human uterus, only 1,4,5-trisphosphate-induced Ca2+ release has been studied extensively. Ryanodine receptor channels are present in myometrium. We determined the role of the cyclic ADP-ribose (cADPR)-signaling pathway in oxytocin-induced intracellular Ca2+ [(Ca2+)i] transients in human myometrial cells. We found that oxytocin-induced Ca2+ transient is dependent on several sources of Ca2+, including extracellular Ca2+ and intracellular Ca2+ stores. In addition, we found that both the 1,4,5-trisphosphate- and the cADPR induced Ca2+ releasing systems are important for the induction of [Ca2+]i transients by oxytocin in human myometrial cells. Furthermore, we investigated TNFalpha regulation of oxytocin-induced [Ca2+]i transients, CD38 cyclase activity, and CD38 expression in human myometrial cells. We found that oxytocin induced [Ca2+]i transients were significantly increased by 50 ng/ml TNF. Similarly, CD38 mRNA levels, CD38 expression, and cyclase activity were increased by TNFalpha, thus increasing cADPR levels. We propose that a complex interaction between multiple signaling pathways is important for the development of intracellular Ca2+ transients induced by oxytocin and that TNFalpha may contribute for the myometrium preparation for labor by regulating the cADPR signaling pathway. The observation that the cADPR-signaling pathway is important for the development of intracellular Ca2+ transients in human myometrial cells raises the possibility that this signaling pathway could serve as a target for the development of new therapeutic strategies for abnormal myometrial contraction observed during pregnancy. PMID- 14563703 TI - Cathepsin L gene expression and promoter activation in rodent granulosa cells. AB - The cysteine protease cathepsin L exhibits hormone-regulated expression during ovulation. In situ hybridization analyses of immature and pregnant mare serum gonadotropin-treated mouse and rat ovaries showed that cathepsin L expression in granulosa cells of small, growing follicles increased in periovulatory follicles after human chorionic gonadotropin stimulation. In the rat ovary, cathepsin L was also expressed in follicles with signs of atresia. To determine the molecular mechanisms that mediate the diverse regulation of this gene in granulosa cells, rat cathepsin L promoter-reporter constructs were analyzed by transient transfection assays in rat granulosa cells and EMSAs. A construct containing the transcriptional start site and -244 bp of upstream promoter sequence (-244/+33 bp) exhibited inducibility by forskolin, the phorbol ester phorbol myristate acetate, and an additive effect of both. Within this region, three functional specificity protein 1 (Sp1) sites, an overlapping early growth response protein-1 site, and a cAMP regulatory element-binding protein site were identified. Single or double mutants of the above-mentioned sites did not alter forskolin/phorbol myristate acetate inducibility of the promoter. Mutation of all three Sp1/specificity protein 3 (Sp3) sites, which also mutated the early growth response protein-1 site, reduced the promoter activation. Mutation of the cAMP regulatory element-binding protein site in the triple Sp1 mutant construct completely blocked the inducibility of the promoter. When these same constructs were transfected into MCF-7 human breast cancer cells or were cotransfected with an Sp1 expression vector in Drosophila SL2 cells, similar results were obtained. Collectively, the data document that three Sp1/specificity protein 3 binding GC rich regions and a functional cAMP regulatory element constitute an important transcriptional regulatory complex for expression of the cathepsin L gene in rat granulosa cells. PMID- 14563704 TI - Localization of ISG15 and conjugated proteins in bovine endometrium using immunohistochemistry and electron microscopy. AB - The interferon-stimulated gene ISG15, a ubiquitin homolog, becomes conjugated to and regulates uterine proteins in response to conceptus-derived interferon-tau on d 18 of pregnancy. It was hypothesized here that cellular localization of ISG15 within endometrial cells might provide insight regarding function. Uteri were collected from cows (approximately 21-d estrous cycle) on d 17-21/0 of the estrous cycle and pregnancy and d 23, 45, and 50 of pregnancy. Intracellular ISG15 and its conjugates were present on d 17 of pregnancy, peaked to highest levels from d 18 to 23 and then declined to low but detectable levels by d 45 (P < 0.05) based on Western blotting. ISG15 and its conjugates were not detected on d 50 of pregnancy or during the estrous cycle. Immunohistochemistry revealed that ISG15 was localized throughout the endometrium on d 18-23, with heaviest staining in the sublumenal stratum compactum and the glandular epithelium throughout the stratum spongiosum. By d 45 and 50, ISG15 was lightly stained only in the stratum compactum immediately beneath the lumenal epithelium. Using transmission electron microscopy and immunogold labeling, ISG15 was specifically localized to organelles and compartments of endometrial epithelial cells and stromal cells: nucleus, perinuclear space, cytosol, mitochondria, rough endoplasmic reticulum, and cell membrane. This specific localization in epithelial and stromal cells led to the conclusion that ISG15 has diverse intracellular functions. The sustained presence of conjugated ISG15 through d 50 of pregnancy might reflect stabilization of conjugated proteins in response to implantation and the development of the placenta. PMID- 14563705 TI - Effects of loss of steroid receptor coactivator-1 on the skeletal response to estrogen in mice. AB - Steroid receptor coactivator (SRC)-1 is an important nuclear receptor coactivator that enhances estrogen (E) action in many tissues, but its role in mediating E effects on bone is unknown. Thus, we assessed the skeletal response to ovariectomy (ovx) and E replacement in SRC-1 knockout (KO) mice compared with wild-type (WT) littermates. Bone mineral density was measured by dual-energy x ray absorptiometry and peripheral quantitative computed tomography at baseline and after 2 months of sham surgery, ovx, or ovx plus E replacement. Microcomputed tomography and bone histomorphometry were also performed at the end of the study. Both WT and SRC-1 KO mice lost bone at multiple sites after ovx; however, although an estradiol (E(2)) dose of 10 microg/kg.d completely prevented loss of cancellous bone (at the lumbar spine and tibial metaphysis) in the WT mice, it was entirely ineffective in preventing cancellous bone loss at these sites in the SRC-1 KO mice. This E(2) dose was, however, equally effective on cortical bone in the tibia in the SRC-1 KO and WT mice. Moreover, a 4-fold higher dose of E(2) was able to overcome the deficit in E action in cancellous bone in the SRC-1 KO mice. These findings establish that, in mice, loss of SRC-1 leads to skeletal resistance to E predominantly in cancellous bone. PMID- 14563706 TI - Cytochrome P450 2D catalyze steroid 21-hydroxylation in the brain. AB - mRNA of cytochrome P450 21-hydroxylase (P450c21) is expressed in the brain, but little is known about the enzymatic properties of P450c21 in the brain. In the present study, we showed, by using various recombinant cytochrome P450 (CYP)2D enzymes and anti-CYP2D4- or P450c21-specific antibodies, that rat brain microsomal steroid 21-hydroxylation is catalyzed not by P450c21, but by CYP2D isoforms. Rat CYP2D4 and human CYP2D6, which are the predominant CYP2D isoforms in the brain, possess 21-hydroxylation activity for both progesterone and 17alpha hydroxyprogesterone. In rat brain microsomes, these activities were not inhibited by anti-P450c21 antibodies, but they were effectively inhibited by the CYP2D specific chemical inhibitor quinidine and by anti-CYP2D4 antibodies. mRNA and protein of CYP2D4 were expressed throughout the brain, especially in cerebellum, striatum, pons, and medulla oblongata, whereas the mRNA and protein levels of P450c21 were extremely low or undetectable. These results support the idea that CYP2D4, not P450c21, works as steroid 21-hydroxylase in the brain. Allopregnanolone, a representative gamma-aminobutyric acid receptor modulator, was also hydroxylated at the C-21 position by recombinant CYP2D4 and CYP2D6. Rat brain microsomal allopregnanolone 21-hydroxylation was inhibited by fluoxetine with an IC(50) value of 2 microm, suggesting the possibility that the brain CYP2D isoforms regulate levels of neurosteroids such as allopregnanolone, and that this regulation is modified by central nervous system-active drugs such as fluoxetine. PMID- 14563707 TI - Adenosine: a modulator of the cardiac response to stress. PMID- 14563708 TI - NHE-1 and NHE-6 activities: ischemic and reperfusion injury. PMID- 14563709 TI - Structure, regulation, and function of mammalian membrane guanylyl cyclase receptors, with a focus on guanylyl cyclase-A. AB - Besides soluble guanylyl cyclase (GC), the receptor for NO, there are at least seven plasma membrane enzymes that synthesize the second-messenger cGMP. All membrane GCs (GC-A through GC-G) share a basic topology, which consists of an extracellular ligand binding domain, a short transmembrane region, and an intracellular domain that contains the catalytic (GC) region. Although the presence of the extracellular domain suggests that all these enzymes function as receptors, specific ligands have been identified for only three of them (GC-A through GC-C). GC-A mediates the endocrine effects of atrial and B-type natriuretic peptides regulating arterial blood pressure and volume homeostasis and also local antihypertrophic actions in the heart. GC-B is a specific receptor for C-type natriuretic peptide, having more of a paracrine function in vascular regeneration and endochondral ossification. GC-C mediates the effects of guanylin and uroguanylin on intestinal electrolyte and water transport and on epithelial cell growth and differentiation. GC-E and GC-F are colocalized within the same photoreceptor cells of the retina and have an important role in phototransduction. Finally, the functions of GC-D (located in the olfactory neuroepithelium) and GC-G (expressed in highest amounts in lung, intestine, and skeletal muscle) are completely unknown. This review discusses the structure and functions of membrane GCs, with special emphasis on the physiological endocrine and cardiac functions of GC-A, the regulation of hormone-dependent GC-A activity, and the relevance of alterations of the atrial natriuretic peptide/GC-A system to cardiovascular diseases. PMID- 14563710 TI - Cardiac failure associated with G6PD deficiency. PMID- 14563711 TI - Inhibition of hydroxymethylglutaryl-coenzyme a reductase reduces Th1 development and promotes Th2 development. AB - Several prospective clinical studies have indicated that hydroxymethylglutaryl coenzyme A reductase inhibitors, statins, prevent cardiovascular events in part through their antiinflammatory properties. Because inflammation is positively and negatively regulated by T helper (Th) 1 cells and Th2 cells, respectively, we examined the effects of statins on the Th polarization in vitro and in vivo. Here we demonstrated that the statins tested, ie, cerivastatin, simvastatin, lovastatin, and atorvastatin, promoted Th2 polarization through both inhibition of Th1 development and augmentation of Th2 development of CD4+ T cells primed in vitro with anti-CD3 antibody and splenic antigen-presenting cells. Cerivastatin exerted most potent effect on modulation of Th1/Th2 development, and the effect was completely abrogated by an addition of mevalonate. Consistent with in vitro experiments, cerivastatin treatment decreased IFN-gamma production of lymph node cells from mice immunized with ovalbumin emulsified in complete Freund's adjuvant, indicating that Th1 development is also suppressed in an in vivo proinflammatory environment. In this murine model, cerivastatin significantly reduced mesangial matrix expansion of glomeruli in the kidney and attenuated proteinuria. The decrease of glomerular sclerosis by cerivastatin treatment was positively related to the suppression of interferon (IFN)-gamma-producing Th1 response in draining lymph node cells. Hence, these findings strongly suggest that statins' inhibition of 3-hydroxy-3-methylglutaryl-coenzyme A reductase regulates Th1/Th2 polarization in vivo and such a mechanism possibly plays a pathophysiological role in immune-related glomerular injury. PMID- 14563712 TI - Heparan sulfate proteoglycan is a mechanosensor on endothelial cells. AB - The objective of this study was to test whether a glycosaminoglycan component of the surface glycocalyx layer is a fluid shear stress sensor on endothelial cells (ECs). Because enhanced nitric oxide (NO) production in response to fluid shear stress is a characteristic and physiologically important response of ECs, we evaluated NOx (NO2- and NO3-) production in response to fluid shear stress after enzymatic removal of heparan sulfate, the dominant glycosaminoglycan of the EC glycocalyx, from cultured ECs. The significant NOx production induced by steady shear stress (20 dyne/cm2) was inhibited completely by pretreatment with 15 mU/mL heparinase III (E.C.4.2.2.8) for 2 hours. Oscillatory shear stress (10+/-15 dyne/cm2) induced an even greater NOx production than steady shear stress that was completely inhibited by pretreatment with heparinase III. Addition of bradykinin (BK) induced significant NOx production that was not inhibited by heparinase pretreatment, demonstrating that the cells were still able to produce abundant NO after heparinase treatment. Fluorescent imaging with a heparan sulfate antibody revealed that heparinase III treatments removed a substantial fraction of the heparan sulfate bound to the surfaces of ECs. In summary, these experiments demonstrate that a heparan sulfate component of the EC glycocalyx participates in mechanosensing that mediates NO production in response to shear stress. The full text of this article is available online at http://www.circresaha.org. PMID- 14563713 TI - Integrin-associated protein binding domain of thrombospondin-1 enhances insulin like growth factor-I receptor signaling in vascular smooth muscle cells. AB - Insulin-like growth factor-I (IGF-I) stimulates vascular smooth muscle cell (SMC) proliferation and migration. The response of smooth muscle cells to IGF-I is determined not only by activation of the IGF-I receptor but also by at least three other transmembrane proteins, alphaVbeta3, integrin-associated protein (IAP), and SHPS-1. This regulation seems to be attributable to their ability to regulate the transfer of SHP-2 phosphatase, a key component of IGF-I signaling. Ligand occupancy of SHPS-1 with IAP is required for the recruitment and transfer of SHP-2 and subsequent signaling in response to IGF-I. The extracellular matrix protein thrombospondin-1 stimulates an increase in the cell proliferation response to IGF-I. Because thrombospondin-1 is a ligand for IAP, we wished to determine whether the enhancing effect of thrombospondin-1 was mediated through IAP binding. To examine the effect of thrombospondin-1 binding to IAP, we used a peptide termed 4N1K derived from the IAP binding site of thrombospondin-1. Preincubation with 4N1K increased IGF-I-stimulated mitogen-activated protein kinase activation and DNA synthesis. This enhancement seemed to be attributable to its ability to increase the duration of IGF-I-stimulated receptor and insulin receptor substrate-1 (IRS-1) phosphorylation. Preincubation with 4N1K delayed IGF I stimulation of SHPS-1 phosphorylation (attributable to an alteration in IAP SHPS-1 interaction), resulting in a delay in SHP-2 recruitment. This delay in SHP 2 transfer seems to account for the increase in the duration of IGF-I receptor phosphorylation and for enhanced downstream signaling. These observations support the conclusion that thrombospondin-1 and IGF-I seem to function coordinately in stimulating smooth muscle proliferation via the thrombospondin-1 interaction with IAP. PMID- 14563714 TI - AT1b receptor predominantly mediates contractions in major mouse blood vessels. AB - In rodents, angiotensin (Ang) II type-1 (AT1) receptors exist as two pharmacologically identical subtypes: AT1a and AT1b. Recent studies have utilized mouse models with specific subtype receptor deletions to differentiate the functional difference between AT1 subtypes. However, little information is available on AT1 subtype expression in mouse vasculature. Therefore, in this study, AT1a-/- mice and wild-type littermates (AT1a+/+) were used to examine AT1 subtype expression and its functional relevance in mouse arterial vessels. Using RT-PCR and restriction enzyme digestion, we showed that AT1b accounts for most of the total AT1 mRNA in mouse abdominal aorta and femoral artery. In contrast, AT1a is the predominant subtype in kidney. To study the functional role of AT1 subtypes, we measured the in vitro contractility in vessels from AT1a-/- and AT1a+/+ mice. The Ang II concentration response curves in abdominal aorta and femoral artery were comparable between the two mouse strains. Furthermore, the Ang II response in AT1a-/- mouse vessels was completely antagonized by losartan, an AT1 antagonist. These results demonstrate that AT1b receptor is a major mediator for Ang II contractile response in mouse vessels, such as abdominal aorta and femoral artery. PMID- 14563715 TI - Site of origin and molecular substrate of atrioventricular junctional rhythm in the rabbit heart. AB - During failure of the sinoatrial node, the heart can be driven by an atrioventricular (AV) junctional pacemaker. The position of the leading pacemaker site during AV junctional rhythm is debated. In this study, we present evidence from high-resolution fluorescent imaging of electrical activity in rabbit isolated atrioventricular node (AVN) preparations that, in the majority of cases (11 out of 14), the AV junctional rhythm originates in the region extending from the AVN toward the coronary sinus along the tricuspid valve (posterior nodal extension, PNE). Histological and immunohistochemical investigation showed that the PNE has the same morphology and unique pattern of expression of neurofilament160 (NF160) and connexins (Cx40, Cx43, and Cx45) as the AVN itself. Block of the pacemaker current, If, by 2 mmol/L Cs+ increased the AV junctional rhythm cycle length from 611+/-84 to 949+/-120 ms (mean+/-SD, n=6, P<0.001). Immunohistochemical investigation showed that the principal If channel protein, HCN4, is abundant in the PNE. As well as the AV junctional rhythm, the PNE described in this study may also be involved in the slow pathway of conduction into the AVN as well as AVN reentry, and the predominant lack of expression of Cx43 as well as the presence of Cx45 in the PNE shown could help explain its slow conduction. PMID- 14563716 TI - Reversal of chronic molecular and cellular abnormalities due to heart failure by passive mechanical ventricular containment. AB - Passive mechanical containment of failing left ventricle (LV) with the Acorn Cardiac Support Device (CSD) was shown to prevent progressive LV dilation in dogs with heart failure (HF) and increase ejection fraction. To examine possible mechanisms for improved LV function with the CSD, we examined the effect of CSD therapy on the expression of cardiac stretch response proteins, myocyte hypertrophy, sarcoplasmic reticulum Ca2+-ATPase activity and uptake, and mRNA gene expression for myosin heavy chain (MHC) isoforms. HF was produced in 12 dogs by intracoronary microembolization. Six dogs were implanted with the CSD and 6 served as concurrent controls. LV tissue from 6 normal dogs was used for comparison. Compared with normal dogs, untreated HF dogs showed reduced cardiomyocyte contraction and relaxation, upregulation of stretch response proteins (p21ras, c-fos, and p38 alpha/beta mitogen-activated protein kinase), increased myocyte hypertrophy, reduced SERCA2a activity with unchanged affinity for calcium, reduced proportion of mRNA gene expression for alpha-MHC, and increased proportion of beta-MHC. Therapy with the CSD was associated with improved cardiomyocyte contraction and relaxation, downregulation of stretch response proteins, attenuation of cardiomyocyte hypertrophy, increased affinity of the pump for calcium, and restoration of alpha- and beta-MHC isoforms ratio. The results suggest that preventing LV dilation and stretch with the CSD promotes downregulation of stretch response proteins, attenuates myocyte hypertrophy and improves SR calcium cycling. These data offer possible mechanisms for improvement of LV function after CSD therapy. PMID- 14563717 TI - Breast feeding and obesity. PMID- 14563718 TI - Intrauterine factors, adiposity, and hyperinsulinaemia. PMID- 14563719 TI - Herbal medicines put into context. PMID- 14563720 TI - Equality for people with disabilities in medicine. PMID- 14563721 TI - When to retract? PMID- 14563724 TI - Second vCJD patient to receive experimental treatment. PMID- 14563725 TI - MEPs lead demand to get sudden unexplained deaths recognised as a syndrome. PMID- 14563730 TI - GP suspended for lying to cover up negligence. PMID- 14563731 TI - Imperial Tobacco plans to deny link between smoking and lung cancer. PMID- 14563732 TI - Specialists clarify when the benefits of HRT outweigh the risks. PMID- 14563734 TI - Murderer can be forced to take medication to become sane enough to be executed. PMID- 14563736 TI - Half of doctors experience violence or abuse from patients. PMID- 14563737 TI - Medical project investigating torture gets under way. PMID- 14563740 TI - Corporate hijacking of food is the most important health hazard of our time. PMID- 14563741 TI - Safety strategy needed in NHS to stop injuries to staff. PMID- 14563743 TI - Montelukast and fluticasone compared with salmeterol and fluticasone in protecting against asthma exacerbation in adults: one year, double blind, randomised, comparative trial. AB - OBJECTIVES: To assess the effect of montelukast versus salmeterol added to inhaled fluticasone propionate on asthma exacerbation in patients whose symptoms are inadequately controlled with fluticasone alone. Design and setting A 52 week, two period, double blind, multicentre trial during which patients whose symptoms remained uncontrolled by inhaled corticosteroids were randomised to add montelukast or salmeterol. PARTICIPANTS: Patients (15-72 years; n = 1490) had a clinical history of chronic asthma for > or = 1 year, a baseline forced expiratory volume in one second (FEV1) value 50-90% predicted, and a beta agonist improvement of > or = 12% in FEV1. MAIN OUTCOME MEASURES: The primary end point was the percentage of patients with at least one asthma exacerbation. RESULTS: 20.1% of the patients in the group receiving montelukast and fluticasone had an asthma exacerbation compared with 19.1% in the group receiving salmeterol and fluticasone; the difference was 1% (95% confidence interval -3.1% to 5.0%). With a risk ratio (montelukast-fluticasone/salmeterol-fluticasone) of 1.05 (0.86 to 1.29), treatment with montelukast and fluticasone was shown to be non-inferior to treatment with salmeterol and fluticasone. Salmeterol and fluticasone significantly increased FEV1 before a beta agonist was used and morning peak expiratory flow compared with montelukast and fluticasone (P < or = 0.001), whereas FEV1 after a beta agonist was used and improvements in asthma specific quality of life and nocturnal awakenings were similar between the groups. Montelukast and fluticasone significantly (P = 0.011) reduced peripheral blood eosinophil counts compared with salmeterol and fluticasone. Both treatments were generally well tolerated. CONCLUSION: The addition of montelukast in patients whose symptoms remain uncontrolled by inhaled fluticasone could provide equivalent clinical control to salmeterol. PMID- 14563744 TI - Prognosis of angina with and without a diagnosis: 11 year follow up in the Whitehall II prospective cohort study. AB - OBJECTIVE: To investigate the prognosis of angina among people with and without diagnosis by a doctor and an abnormal cardiovascular test result. DESIGN: Prospective cohort study with a median follow up of 11 years. SETTING: 20 civil service departments originally located in London. PARTICIPANTS: 10 308 civil servants aged 35-55 years at baseline. MAIN OUTCOME MEASURES: Recurrent reports of angina; quality of life (SF-36 physical functioning); non-fatal myocardial infarction; death from any cause (n = 344). RESULTS: 1158 (11.4%) participants developed angina, and 813 (70%) had no evidence of diagnosis by a doctor at the time of the initial report. Participants without a diagnosis had an increased risk of impaired physical functioning (age and sex adjusted odds ratio of 2.36 (95% confidence interval 1.91 to 2.90)) compared with those who had neither angina nor myocardial infarction throughout follow up. Among reported cases of angina without a diagnosis, the 15.5% with an abnormality on a study electrocardiogram had an increased risk of death (hazard ratio 2.37 (1.16 to 4.87)). These effects were similar in magnitude to those in participants with a diagnosis of angina. CONCLUSION: Undiagnosed angina was common and had an adverse impact on prognosis comparable to that of diagnosed angina, particularly among people with electrocardiographic abnormalities. Efforts to improve prognosis among people with angina should take account of this submerged clinical iceberg. PMID- 14563745 TI - Effects of grandmothers' smoking in pregnancy on birth weight: intergenerational cohort study. AB - OBJECTIVE: To investigate the influences on birth weight of maternal smoking during pregnancy across generations. DESIGN: Intergenerational cohort study. PARTICIPANTS: Members of the 1958 birth cohort and their offspring and mothers. SETTING: England, Scotland, and Wales. MAIN OUTCOME MEASURE: Birth weight. RESULTS: Information on grandmothers' smoking during pregnancy was available for 9028 singleton offspring of 4302 female cohort members. Assuming heritable transmission through the intergenerational association, grandmothers' smoking was predicted to result in a 34 g reduction (95% confidence interval -41 g to -28 g) in the birth weight of grandchildren. Random effects models showed a negative association between grandmothers' smoking and birth weight of grandchildren (beta regression coefficient -24 g, -50 g to 3 g), but this effect was eliminated after adjustment for maternal smoking (0 g, -26 g to 26 g). No association was evident among the offspring of non-smoking mothers (n = 6105; 14 g, -17 g to 46 g), and after adjustment for maternal birth weight, adult height and body mass index, grandmothers' smoking was positively associated with the birth weight of grandchildren (45 g, 10 g to 80 g). CONCLUSION: Deficits in mothers' birth weight attributable to their mother smoking was not evident in the grandchildren. PMID- 14563746 TI - Anthropometry and body composition of 18 year old men according to duration of breast feeding: birth cohort study from Brazil. AB - OBJECTIVE: To assess the association between duration of breast feeding and measures of adiposity in adolescence. DESIGN: Population based birth cohort study. SETTING: Pelotas, a city of 320 000 inhabitants in a relatively developed area in southern Brazil. PARTICIPANTS: All newborn infants in the city's hospitals were enrolled in 1982; 78.8% (2250) of all male participants were located at age 18 years when enrolling in the national army. MAIN OUTCOME MEASURES: Weight, height, sitting height, subscapular and triceps skinfolds, and body composition (body fat, lean mass). RESULTS: Neither the duration of total breast feeding nor that of predominant breast feeding (breast milk plus non nutritive fluids) showed consistent associations with anthropometric or body composition indices. After adjustment for confounding factors, the only significant associations were a greater than 50% reduction in obesity among participants breast fed for three to five months compared with all other breastfeeding categories (P = 0.007) and a linear decreasing trend in obesity with increasing duration of predominant breast feeding (P = 0.03). Similar significant effects were not observed for other measures of adiposity. Borderline direct associations also occurred between total duration of breast feeding and adult height (P = 0.06). CONCLUSIONS: The significant reduction in obesity among children breast fed for three to five months is difficult to interpret, as no a priori hypothesis existed regarding a protective effect of intermediate duration of breast feeding. The findings indicate that, in this population, breast feeding has no marked protective effect against adolescent adiposity. PMID- 14563747 TI - Breast feeding and obesity in childhood: cross sectional study. PMID- 14563748 TI - Drug points: Weight loss associated with levetiracetam. PMID- 14563749 TI - Effectiveness of out-of-home day care for disadvantaged families: randomised controlled trial. AB - OBJECTIVE: To assess the effects of providing daycare facilities for young children on the health and welfare of disadvantaged families. DESIGN: Randomised controlled trial. Eligible children from the application list to a daycare facility were randomly allocated to receive a daycare place or not. SETTING: Early Years daycare centre in Borough of Hackney, London. PARTICIPANTS: 120 mothers and 143 eligible children (aged between 6 months and 3.5 years). INTERVENTION: A place at the centre, which provided high quality day care. Control families used other child care that they secured for themselves. MAIN OUTCOME MEASURES: Maternal paid employment, household income, child health and development. RESULTS: At 18 months' follow up, 67% of intervention group mothers and 60% of control group mothers were in paid employment (adjusted risk ratio 1.23 (95% confidence interval 0.99 to 1.52)), but were no more likely to have a weekly household income of above pound 200 (risk ratio 0.88 (0.70 to 1.09)). Intervention group children had more otitis media with effusion (risk ratio 1.74 (1.02 to 2.96)) and used more health services (1.58 (1.05 to 2.38)), but both estimates were imprecise. CONCLUSION: The provision of child day care may have increased maternal employment, but it did not seem to increase household income. The results suggest that providing day care may be insufficient as a strategy to reduce poverty. The study shows how random allocation can be used to ration and evaluate interventions where demand exceeds supply. PMID- 14563751 TI - Recent developments in vascular surgery. PMID- 14563759 TI - Cochrane at crossroads over drug company sponsorship. PMID- 14563760 TI - Recruiting doctors from poor countries: the great brain robbery? PMID- 14563758 TI - Assisted conception. III--problems with assisted conception. PMID- 14563761 TI - Recruitment is ethical. PMID- 14563762 TI - Third world brain drain: performance of health professionals and systems needs to be assessed. PMID- 14563763 TI - Retraction of paper on maternal diabetes. PMID- 14563764 TI - Third world brain drain: causes of exodus need to be examined and rectified. PMID- 14563765 TI - Breast cancer in men: group is exploring issues for men with breast cancer across the United Kingdom. PMID- 14563767 TI - Third world brain drain: brain drain must be halted. PMID- 14563766 TI - Breast cancer in men: evidence suggests preoperative work up is suboptimal. PMID- 14563768 TI - Third world brain drain: don't punish the children. PMID- 14563769 TI - Consistency in anaesthetic care: segmentation of anaesthetic care is not inherently bad. PMID- 14563770 TI - Consistency in anaesthetic care: patients' attitudes matter. PMID- 14563771 TI - Rate and rhythm are important in critically ill patients. PMID- 14563772 TI - Consistency in anaesthetic care: European practice may not be relevant to the United Kingdom. PMID- 14563773 TI - Tuberculin testing before BCG vaccination: effect of age on adverse effects has not been made clear. PMID- 14563774 TI - What happened to evidence in NICE guidance on heart failure? PMID- 14563775 TI - General practitioners with special clinical interests: experience has been successful. PMID- 14563776 TI - Might money spent on statins be better spent? PMID- 14563777 TI - Is there an identifiable doctor on board? PMID- 14563778 TI - General practitioners with special clinical interests: evidence for effectiveness exists. PMID- 14563779 TI - Organising international humanitarian and health work. PMID- 14563780 TI - The General Medical Council and medical students with disabilities. PMID- 14563782 TI - Hyperosmolar solution effects in guinea pig airways. I. Mechanical responses to relative changes in osmolarity. AB - In the guinea pig isolated perfused trachea contracted with serosal methacholine (MCh), increasing the osmolarity of the mucosal bathing solution elicits relaxation of smooth muscle mediated by epithelium-derived relaxing factor (EpDRF). The present study was undertaken to determine whether a specific modality of the hyperosmolar stimulus induced the relaxation response. Mucosal hyperosmolar challenge with D-mannitol, N-methyl-D-glucamine (NMDG)-chloride, NMDG-gluconate (NMDG-Glu), or urea elicited relaxation with equal potency. In contrast, hyperosmolar solutions at the serosal surface induced diverse, osmolyte specific responses. In tracheae contracted with MCh, abrupt replacement of the mucosal modified Krebs-Henseleit solution (MKHS) with isosmolar osmolyte solutions to stimulate cell shrinkage elicited five discrete response patterns related to the membrane permeance of the solute, but increasing the osmolarity of the isosmolar solution via the further addition of the same solute always induced relaxation. Similarly, perfusion of the lumen with water induced a transient contraction, but subsequent addition of MKHS, or isosmolar D-mannitol, urea, NMDG Glu, NaCl, or KCl induced relaxation. Subsequent hyperosmolar addition of the same osmolyte-evoked relaxation. Compatible osmolytes had no effect on smooth muscle tone and did not affect responses to hyperosmolar challenge. The results suggest that the airway epithelium acts as an osmolarity sensor, which communicates with airway smooth muscle through EpDRF. The mechanical responses of the smooth muscle resulting from changes in the osmotic environment are associated with discrete modalities of the osmolar stimulus, including membrane reflection of the particles, incremental change in osmolarity and directionality, but not cell shrinkage. PMID- 14563783 TI - Modification by dopaminergic drugs of choice behavior under concurrent schedules of intravenous saline and food delivery in monkeys. AB - The allocation of "choice" behavior provides a measure that may be useful in developing experimental models of clinical relapse. In the present experiments, indirect monoaminergic agonists [cocaine, 1-(2-[bis(4-fluorophenyl)methoxy]ethyl) 4-(3-phenylpropyl)piperazine (GBR 12909), desipramine, and citalopram], and dopaminergic D1 family agonists [(+/-)-6-chloro-7,8-dihydroxy-3-allyl-1-phenyl 2,3,4,5-tetrahydro-1H-3-benzazepine (SKF 82958), R-(+)-6-bromo-7,8-dihydroxy-3 allyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine (R-(+)-6-BrAPB), and 6-chloro 7,8-dihydroxy-3-methyl-1-(3-methylphenyl)-2,3,4,5-tetrahydro-1H-3-benzazepine (SKF 83959)] and D2 family agonists [quinelorane, R-(-)-10,11-dihydroxy-N-n propylnorapomorphine (R-NPA), (+)-N-propyl-hydroxynaphoxazine [(+)-PHNO], and S (+)-(4aR,10bR)-3,4,4a,10b-tetrahydro-4-propyl-2H,5H-[1]benzopyrano-[4,3-b]-1,4 oxazin-9-ol (PD 128907)] were evaluated for their capacity to alter the distribution of choice behavior in cocaine-experienced monkeys. Rhesus monkeys responded on two levers (injection-lever and food-lever) under concurrent fixed ratio 30; fixed ratio 30 schedules of i.v. cocaine and food delivery. Under training conditions, the distribution of behavior was related to the unit dose of i.v. cocaine: when saline was available, responding occurred predominantly on the food-lever and when reinforcing doses of cocaine were available, responding occurred predominantly on the injection-lever. Drugs were studied by administering i.m. pretreatment doses before components in sessions of i.v. saline availability. Cocaine produced dose-related increases in injection-lever responding in all monkeys, whereas desipramine failed to alter the distribution of behavior in any monkey. The dopamine transport blocker GBR 12909 and each dopamine D1 family agonist markedly increased injection-lever responding in three of four monkeys; the serotonin transport blocker citalopram and D2 family agonists were comparably effective in only one or two monkeys. These results agree with previous findings of similarity in the behavioral effects of cocaine and indirect or direct dopamine agonists and suggest, furthermore, that i.v. self administration behavior engendered by priming doses of cocaine may involve actions mediated through both D1 and D2 families of dopamine receptors. PMID- 14563784 TI - Muscarinic agonist-mediated heterologous desensitization in isolated ileum requires activation of both muscarinic M2 and M3 receptors. AB - We investigated the subtypes of the muscarinic receptor mediating short-term heterologous desensitization in the isolated ileum. Treatment of the ileum from C57BL/6 mice with acetylcholine (30 microM) for 20 min caused a subsequent decrease in contractile sensitivity to both prostaglandin F2alpha (PGF2alpha) and the muscarinic agonist, oxotremorine-M. This subsensitivity was characterized by 7- and 3-fold increases in the EC50 values of the agonists, respectively, with no significant effect on the maximal response. The subsensitivity to PGF2alpha was prevented in both M2 and M3 muscarinic receptor knockout mice. Similarly, the subsensitivity to oxotremorine-M was prevented in M2 knockout mice. Acetylcholine mediated desensitization of histamine-induced contractions in the guinea pig ileum was inhibited by both M2- and M3-selective muscarinic antagonists with high potency, although careful analysis of the data suggested behavior more consistent with an M2 antagonistic profile. Modeling studies showed that the competitive antagonism of response contingent upon activation of two receptor subtypes should exhibit a pharmacological profile similar to that of the least sensitive signaling pathway. Our results demonstrate that muscarinic agonist-mediated short term heterologous desensitization of intestinal smooth muscle is contingent upon activation of both M2 and M3 muscarinic receptors and that activation of either receptor by itself is insufficient to cause desensitization. PMID- 14563785 TI - Nicotinic receptor modulation of dopamine transporter function in rat striatum and medial prefrontal cortex. AB - Nicotine activates nicotinic acetylcholine receptors (nAChRs) on dopamine (DA) terminals to evoke DA release, which subsequently is taken back up into the terminal via the DA transporter (DAT). nAChRs may modulate DAT function thereby contributing to the regulation of synaptic DA concentrations. The present study determined the dose-response for nicotine (0.1-0.8 mg/kg, s.c.) to modulate DA clearance in striatum and medial prefrontal cortex (mPFC) using in vivo voltammetry in urethane anesthetized rats and determined if this effect was mediated by nAChRs. Exogenous DA (200 microM) was pressure-ejected at 5-min intervals until reproducible baseline signals were obtained. Subsequently, nicotine or saline was administered, and DA pressure ejection continued at 5-min intervals for 60 min. In both striatum and mPFC, signal amplitude decreased by approximately 20% across the 60-min session in saline-injected rats. A monophasic dose-response curve was found in striatum, with a maximal 50% decrease in signal amplitude after 0.8 mg/kg. In contrast, a U-shaped dose-response curve was found in mPFC, with a maximal 50% decrease in signal amplitude after 0.4 mg/kg. Onset of nicotine response occurred 10 to 15 min after injection in both brain regions; however, the amount of time before maximal response was 45 and 30 min in striatum and mPFC, respectively. Mecamylamine (1.5 mg/kg) completely inhibited the nicotine-induced (0.8 and 0.4 mg/kg) decrease in signal amplitude in striatum and mPFC, respectively, indicating mediation by nAChRs. Thus, nicotine enhances DA clearance in striatum and mPFC in a mecamylamine-sensitive manner, indicating that nAChRs modulate DAT function in these brain regions. PMID- 14563786 TI - Chronopharmacology of melatonin in mice to maximize the antitumor effect and minimize the rhythm disturbance effect. AB - The influence of dosing time on the antitumor effect and the rhythm disturbance effect of melatonin (MLT) was investigated in ICR male mice under a light/dark (12:12) cycle. In tumor-bearing mice, the antitumor effect of MLT (1 mg/kg intraperitoneal) was most effective in the dark phase; and the rhythm disturbance effect of MLT on the locomotor activity was more serious in the light phase than in the dark phase. The antitumor effect and the rhythm disturbance effect of MLT increased when the specific binding of MLT receptor in target tissues, tumor or suprachiasmatic nucleus, increased and they decreased when the level decreased. Furthermore, because luzindole, an MT1 and MT2 blocker, caused the antitumor effect or rhythm disturbance effect of MLT to decrease, it is suggested that the time-dependent change of the pharmacological effects of MLT were influenced by that of MLT receptor(s) function. On the other hand, there was no significant dosing time-dependent change of MLT concentration in tumor or brain after injection. Thus, the pharmacokinetic factor does not seem to contribute to the dosing time-dependent effect of MLT. These results suggest that by choosing the most suitable dosing time for MLT, the efficacy of the drug can be increased, and the toxicity of the drug can be decreased in certain experimental and clinical situations. PMID- 14563787 TI - Six novel nonsynonymous CYP1A2 gene polymorphisms: catalytic activities of the naturally occurring variant enzymes. AB - Six novel nonsynonymous nucleotide alterations were found in the cytochrome P450 1A2 gene in a Japanese population, which resulted in the following amino acid substitutions: T83M, E168Q, F186L, S212C, G299A, and T438I. These individuals were heterozygous for the amino acid substitutions. The potential functional alterations caused by the amino acid substitutions were characterized by a cDNA mediated expression system using Chinese hamster V79 cells. Among the six CYP1A2 variants, F186L showed the most profound and statistically significant reduction in O-deethylation of phenacetin and 7-ethoxyresorufin. Kinetic analyses performed for the ethoxyresorufin O-deethylation revealed that the Vmax of the F186L variant was approximately 5% of that of the CYP1A2 wild type, despite a 5-fold lower Km value of the variant, the consequence of which was reduced enzymatic activity toward the substrate. Thus, for the first time, phenylalanine at residue 186 is suggested to be a critical amino acid for catalytic activity. PMID- 14563788 TI - Antinociceptive effects of novel A2B adenosine receptor antagonists. AB - Caffeine, an adenosine A1, A2A, and A2B receptor antagonist, is frequently used as an adjuvant analgesic in combination with nonsteroidal anti-inflammatory drugs or opioids. In this study, we have examined the effects of novel specific adenosine receptor antagonists in an acute animal model of nociception. Several A2B-selective compounds showed antinociceptive effects in the hot-plate test. In contrast, A1- and A2A-selective compounds did not alter pain thresholds, and an A3 adenosine receptor antagonist produced thermal hyperalgesia. Evaluation of psychostimulant effects of these compounds in the open field showed only small effects of some antagonists at high doses. Coadministration of low, subeffective doses of A2B-selective antagonists with a low dose of morphine enhanced the efficacy of morphine. Our results indicate that analgesic effects of caffeine are mediated, at least in part, by A2B adenosine receptors. PMID- 14563789 TI - Roles of superoxide, peroxynitrite, and protein kinase C in the development of tolerance to nitroglycerin. AB - A current hypothesis states that tolerance to nitroglycerin (GTN) involves increased formation of superoxide (O2*-). Studies showing that inhibitors of protein kinase C (PKC) prevent tolerance to GTN suggest the involvement of PKC activation, which can also increase O2*-. We examined the roles of O2*-, peroxynitrite (ONOO-), and PKC activation in GTN tolerance. Pre-exposure of rat aortic rings to GTN (5 x 10(-4) M) for 2 h caused tolerance to the vasodilating effect of GTN, as evidenced by a substantial rightward shift of GTN concentration relaxation curves. This shift was reduced by treatment of the rings with the antioxidants uric acid, vitamin C, or tempol or the PKC inhibitor chelerythrine. We also found that O2*- generation via xanthine/xanthine oxidase in the bath induced tolerance to GTN. However, responses to nitroprusside were not affected. In vivo tolerance produced in rats by 3-day i.v. infusion of GTN was also almost completely prevented by coinfusion of tempol. In bovine aortic endothelial cells (EC), addition of GTN produced a marked increase in tyrosine nitrosylation, indicating increased ONOO- formation. This action was blocked by prior treatment with uric acid, superoxide dismutase, NG-nitro-L-arginine methyl ester, or chelerythrine. We also demonstrated that GTN translocates the alpha- and epsilonPKC isoforms in EC. However, PKCzeta was not affected by GTN treatment. In conclusion, tolerance to GTN involves enhanced production of O2*- and ONOO- and activation of NO synthase. Furthermore, sustained activation of alpha- and epsilonPKC isozymes in EC by GTN may play a role in development of tolerance. PMID- 14563790 TI - Potent mechanism-based inhibition of human CYP2B6 by clopidogrel and ticlopidine. AB - The thienopyridine derivatives ticlopidine and clopidogrel are inhibitors of ADP induced platelet aggregation. Pharmacological activity of these prodrugs depends on cytochrome P450 (P450)-dependent oxidation to the active antithrombotic agent. In this study, we investigated the interaction potential of clopidogrel and ticlopidine by using human liver microsomes and recombinantly expressed P450 isoforms. Both clopidogrel and ticlopidine inhibited CYP2B6 with highest potency and CYP2C19 with lower potency. Clopidogrel also inhibited CYP2C9, and ticlopidine also inhibited CYP1A2, with lower potency. Inhibition of CYP2B6 was time- and concentration-dependent, and as shown by dialysis experiments, it was irreversible and dependent on NADPH, suggesting a mechanism-based mode of action. Inactivation was of nonpseudo-firstorder type with maximal rates of inactivation (K(inact)) for clopidogrel and ticlopidine in microsomes (recombinant CYP2B6) of 0.35 (1.5 min(-1)) and 0.5 min(-1) (0.8 min(-1)), respectively, and half-maximal inactivator concentrations (KI) were 0.5 microM (1.1 microM) for clopidogrel and 0.2 microM (0.8 microM) for ticlopidine. Inhibition was attenuated by the presence of alternative active site ligands but not by nucleophilic trapping agents or reactive oxygen scavengers, further supporting mechanism-based action. A chemical mechanism is discussed based on the known metabolic activation of clopidogrel and on the finding that hemoprotein integrity of recombinant CYP2B6 was not affected by irreversible inhibition. These results suggest the possibility of drug interactions between thienopyridine derivates and drug substrates of CYP2B6 and CYP2C19. PMID- 14563791 TI - Metacarpophalangeal joint arthroplasty in rheumatoid arthritis. A long-term assessment. AB - BACKGROUND: The long-term results of silicone metacarpophalangeal arthroplasty in patients with rheumatoid arthritis are uncertain. The purpose of this investigation was to evaluate the subjective, objective, and radiographic outcomes at the time of long-term follow-up. METHODS: Patients with rheumatoid arthritis who underwent simultaneous silicone metacarpophalangeal joint arthroplasties of all four fingers by one surgeon were eligible for inclusion in the study. The results of a total of 208 arthroplasties in fifty-two hands of thirty-six patients were evaluated at an average of fourteen years postoperatively. Active metacarpophalangeal joint motion, ulnar drift, and radiographs were assessed. The radiographs were reviewed for changes in bone length, erosions, and implant fractures. The Michigan Hand Outcomes Questionnaire (MHQ) was administered to the patients. RESULTS: The mean arc of motion of the metacarpophalangeal joints improved from 30 degrees preoperatively to 46 degrees immediately after the surgery but decreased to 36 degrees at the time of final follow-up. The mean extension deficit of the metacarpophalangeal joints improved from 57 degrees preoperatively to 11 degrees immediately after the surgery but worsened to 23 degrees at the time of final follow-up. The mean ulnar drift improved from 26 degrees preoperatively to <5 degrees in the immediate postoperative period and then recurred to an average of 16 degrees at the time of final follow-up. Implant fractures were associated with increased ulnar drift (p < 0.001). Bone reaction adjacent to the implant was demonstrated by bone shortening in most patients and by erosions in 29% of the patients. One hundred and thirty implants (63%) were broken and forty-five (22%) more were deformed at the time of final follow-up. The MHQ score averaged 48 of 100 points. The patients expressed satisfaction with the function of only 38% of the hands, and only 27% of the hands were pain-free at the time of final follow-up. A greater degree of ulnar drift was associated with decreased patient satisfaction and a decreased score for the cosmetic appearance (p 2 mm in width. The average postoperative femorotibial alignment was 5 degrees of valgus. Nine knees had progression of arthritis in the unresurfaced compartment; none of these knees were revised, and none of the patients had deterioration in the Hospital for Special Surgery score. Kaplan-Meier analysis demonstrated an eleven-year survivorship of 92%. CONCLUSIONS: At an average duration of follow-up of eleven years, unicompartmental knee arthroplasty was associated with pain relief and excellent function in a cohort of patients who had been sixty years of age or younger and active at the time of surgery. PMID- 14563807 TI - Outcome of surgical treatment of medial tibial stress syndrome. AB - BACKGROUND: Medial tibial stress syndrome is a common chronic sports injury characterized by exercise-induced pain along the posteromedial border of the tibia. The reported outcomes of surgical treatment of this condition have varied. METHODS: Of seventy-eight patients who underwent surgery for medial tibial stress syndrome, forty-six (thirty-one men and fifteen women) returned for follow-up. The outcomes of the surgery were determined by comparing preoperative and postoperative pain levels as indicated on a visual analog pain scale and ascertaining the ability of the athletes to return to presymptom levels of exercise. RESULTS: The mean duration of postoperative follow-up was thirty months (range, six to sixty-three months). Surgery significantly reduced pain levels (p < 0.001) by an average of 72% as indicated on the visual analog pain scale. An excellent result was achieved in 35% of the limbs; a good result, in 34%; a fair result, in 22%; and a poor result, in 9%. Despite the success with regard to pain reduction, for a variety of reasons only nineteen (41%) of the athletes fully returned to their presymptom sports activity. CONCLUSIONS: Surgery can significantly reduce the pain associated with medial tibial stress syndrome. Despite this reduction in pain, athletes should be counseled that a full uninhibited return to sports is not always achieved. PMID- 14563808 TI - Comparison of simultaneous bilateral with unilateral total knee arthroplasty in terms of perioperative complications. AB - BACKGROUND: Previous studies have demonstrated an increased rate of perioperative complications and morbidity following simultaneous bilateral total knee arthroplasty compared with the rate following unilateral total knee arthroplasty. The purpose of this study was to compare the rate of perioperative complications and morbidity associated with simultaneous bilateral total knee arthroplasty with that associated with unilateral total knee arthroplasty. METHODS: The records on all bilateral total knee arthroplasties performed between January 1994 and June 2000 and unilateral total knee arthroplasties performed between January 1995 and June 2000 were retrospectively reviewed. The records on 514 unilateral total knee arthroplasties and 255 bilateral total knee arthroplasties were analyzed to determine demographic information, preoperative comorbidities, perioperative complications, and thirty-day and one-year mortality rates. RESULTS: The rates of some perioperative complications, including myocardial infarction, postoperative confusion, and the need for intensive monitoring, were greater after the bilateral arthroplasties. However, the thirty-day and one-year mortality rates and the risks of pulmonary embolism, infection, and deep venous thrombosis were similar for the two groups. CONCLUSIONS: The risk of perioperative complications associated with bilateral simultaneous total knee arthroplasty was slightly increased compared with that associated with unilateral total knee arthroplasty, but the mortality rates were similar. Ultimately, the decision to proceed with simultaneous knee replacement should depend on patient preference through informed choice. PMID- 14563809 TI - Assessment of early osteoarthritis in hip dysplasia with delayed gadolinium enhanced magnetic resonance imaging of cartilage. AB - BACKGROUND: The efficacy of surgical and medical treatment of osteoarthritis is difficult to assess because of the lack of a noninvasive, sensitive measure of cartilage integrity. Delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC) was designed to specifically examine glycosaminoglycan changes in articular cartilage that occur during the development of osteoarthritis. Our primary goal was to compare this technique with measurement of the joint space width on conventional radiographs in patients with hip dysplasia. We performed this comparison by assessing the correlation between the findings of each technique and clinically important factors such as pain, severity of dysplasia, and age. METHODS: Sixty-eight hips in forty-three patients were included in the study. Clinical symptoms were assessed with use of the Western Ontario and McMaster Universities Osteoarthritis (WOMAC) questionnaire. The width of the joint space as well as the lateral center-edge angle of Wiberg (as a measure of the severity of the dysplasia) was measured on standard standing radiographs. Magnetic resonance imaging maps of glycosaminoglycan distribution were made with T1-calculated images after administration of gadopentetate (2-) (Gd-DTPA (2-) ). The dGEMRIC index was calculated as the average of the T1 values for the acetabular and femoral head cartilages. RESULTS: The dGEMRIC index correlated with both pain (rs = -0.50, p < 0.0001) and the lateral center-edge angle (rs = 0.52, p < 0.0001), whereas the joint space width did not correlate with either, with the numbers available. There was a correlation between the dGEMRIC index and pain whether or not a labral tear was present. The dGEMRIC index was significantly different (p < 0.0001) among three groups of hips classified according to whether they had mild, moderate, or severe dysplasia, whereas the joint space width did not differ significantly among these three groups. There was no significant correlation between age and any of the other parameters. CONCLUSIONS: We demonstrated that, in patients with hip dysplasia, the dGEMRIC index-a measure of the biochemical integrity of cartilage-correlates with pain and the severity of the dysplasia and is significantly different among groups of hips with mild, moderate, and severe dysplasia, suggesting that it may be a sensitive measure of early osteoarthritis. Additional studies are needed to determine whether dGEMRIC can be used to predict disease progression in different situations and/or demonstrate responses to therapeutic interventions. PMID- 14563810 TI - Simultaneous revision and contralateral primary total knee arthroplasty. AB - BACKGROUND: There is controversy regarding whether simultaneous or staged bilateral total knee arthroplasty should be performed in patients with bilateral gonarthrosis. In addition, revision total knee arthroplasties have been less successful than primary arthroplasties. The purpose of this study was to evaluate the results of simultaneous revision and contralateral primary total knee arthroplasties performed during the same setting. METHODS: The study cohort included 150 knees in seventy-five patients who had undergone revision arthroplasty because of aseptic failure of a total knee arthroplasty and a contralateral primary arthroplasty for severe gonarthrosis under the same anesthetic. The study group was compared with a control group of sixty patients who had severe arthritis in one knee and had undergone unilateral revision total knee arthroplasty on the contralateral side. The duration of follow-up averaged five years in the group treated with the simultaneous arthroplasties and eight years in the control group. Clinical and radiographic results were evaluated with the Knee Society rating system. RESULTS: In the study group, the average knee score improved by 48 points on the side of the primary arthroplasty and by 37 points on the side of the revision. At the last follow-up visit, most (sixty five) of the seventy-five patients stated that the knee with the revision felt better than the knee with the simultaneously performed primary arthroplasty. In the control group, the average knee score improved by 30 points on the side of the revision arthroplasty. However, forty-five of the sixty patients had a primary total knee arthroplasty on the contralateral side within two years after the index revision arthroplasty; by four years, all sixty patients had undergone a contralateral primary arthroplasty. CONCLUSIONS: We found a favorable outcome in patients who had undergone simultaneous revision and contralateral primary total knee arthroplasties. Despite a lower mean knee score and less motion, most patients seemed to prefer the knee with the revision arthroplasty to the knee with the primary procedure. These results suggest that this combined procedure is a safe and favorable alternative to a staged procedure consisting of revision and subsequent contralateral primary total knee arthroplasty. PMID- 14563811 TI - Osteonecrosis following isolated avulsion fracture of the greater trochanter in children. A report of two cases. PMID- 14563812 TI - Suture granuloma mimicking infection following total hip arthroplasty. A report of three cases. PMID- 14563813 TI - Painful osteoporotic vertebral fracture. Pathogenesis, evaluation, and roles of vertebroplasty and kyphoplasty in its management. PMID- 14563815 TI - One more turn of the wrench. PMID- 14563816 TI - Recombinant human bone morphogenetic protein-2 for treatment of open tibial fractures. PMID- 14563817 TI - Increased frequency of acute local reaction to intra-articular Hylan G-F 20 (Synvisc) in patients receiving more than one course of treatment. PMID- 14563818 TI - Granulomatous inflammation after Hylan G-F 20 viscosupplementation of the knee. PMID- 14563819 TI - What's new in orthopaedic research. PMID- 14563821 TI - A method of direct measurement for the enzymatic determination of cholesteryl esters. AB - A direct measurement method for the enzymatic determination of cholesteryl esters (CEs) without measuring total cholesterol (TC) and free cholesterol (FC) is described. In the first step, hydrogen peroxide generated by cholesterol oxidase from FC was decomposed by catalase. In the second step, CE was measured by enzymatic determination using a colorimetric method or a fluorometric method. The measurement sensitivity of the fluorometric method was more than 20 times that of the colorimetric method. Optimal conditions of the assay were determined, and examples of measured CE in human plasma, rat liver, and cultured cells are indicated. The method of directly measuring CE was simple and has exceptional reproducibility compared with the technique of subtracting FC from TC using each measured TC and FC. PMID- 14563822 TI - Regulation of sterol carrier protein gene expression by the forkhead transcription factor FOXO3a. AB - The SCP gene encodes two proteins, sterol carrier protein X (SCPx) and SCP2, that are independently regulated by separate promoters. SCPx has been shown to be the thiolase involved in the breakdown of branched-chain fatty acids and in the biosynthesis of bile acids. The in vivo function of SCP2 however remains to be established. The transcriptional regulation of SCPx and SCP2 is unclear, but their promoter regions contain several putative regulatory domains. We show here that both SCPx and SCP2 are upregulated by the daf-16-like Forkhead transcription factor FOXO3a (also known as FKHRL1) on the level of promoter activity. It was recently described that Forkheads regulate protection against (oxidative) stress in both Caenorhabditis elegans and mammalian cells. We looked into a role for SCP2 in the cellular defense against oxidative damage and found that a fluorescent fatty acid analog bound to SCP2 is protected against H2O2/Cu2+ induced oxidative damage. We propose a model for the way in which SCP2 could protect fatty acids from peroxidation. PMID- 14563823 TI - Rate of gastric emptying influences dietary cholesterol absorption efficiency in selected inbred strains of mice. AB - This study compared the physiological process of cholesterol absorption in different strains of inbred mice with the goal of identifying novel mechanism(s) by which cholesterol absorption can be controlled. The rate and amount of cholesterol absorption were evaluated based on [14C]cholesterol appearance in plasma after feeding a meal containing [14C]cholesterol and by the percentage of [14C]-cholesterol absorbed over a 24 h period. Results showed that the rate of [14C]cholesterol appearance in plasma was slower in 129P3/J mice than in SJL/J mice. However, more dietary cholesterol was absorbed over a 24 h period by 129P3/J mice than by SJL/J mice. In both strains of mice, cholesterol delivered with medium-chain triglyceride was absorbed less efficiently than cholesterol delivered with olive oil. The strain- and vehicle-dependent differences in cholesterol absorption efficiency correlated negatively with stomach-emptying rates. Furthermore, inhibition of gastric emptying with nitric oxide synthase inhibitor increased cholesterol absorption efficiency in SJL/J mice. These results document that stomach-emptying rate contributes directly to the rate of dietary cholesterol absorption, which is inversely correlated with the total amount of cholesterol absorbed from a single meal. Additionally, genetic factor(s) that influence gastric emptying may be an important determinant of cholesterol absorption efficiency. PMID- 14563824 TI - Regulation of apoA-I gene expression: mechanism of action of estrogen and genistein. AB - We have previously shown that 17-beta-estradiol (E2) and genistein increase the expression of apolipoprotein A-I (apoA-I), the major protein component of HDL, in Hep G2 cells. To elucidate the mechanism mediating the increase in apoA-I gene expression by these compounds, plasmid constructs containing serial deletions of the apoA-I promoter region were generated. The smallest region maintaining response to E2 and genistein spanned the -220 to -148 sequence, and the estrogen antagonist ICI182,780 completely inhibited the E2 and genistein effect. Nuclear extracts from cells treated with E2 and genistein showed increased binding to site B oligonucleotide (-169 to -146), and nuclear extracts from genistein treated cells showed increased binding to an early growth response factor 1 (Egr 1) oligonucleotide compared to control cells. An increase in the concentrations of Egr-1 and hepatocyte nuclear factor-3beta was observed in nuclear extracts of cells treated with both compounds compared to control cells. Treatment with a specific inhibitor of mitogen-activated protein (MAP) kinase, but not with other inhibitors, abolished the stimulation of apoA-I gene expression by E2 and genistein. These results indicate that the MAP kinase pathway is involved in the regulation of apoA-I gene expression by genistein and E2, possibly through downstream regulation of transcription factors binding to the promoter region. PMID- 14563825 TI - Impaired expression of NADH dehydrogenase subunit 1 and PPARgamma coactivator-1 in skeletal muscle of ZDF rats: restoration by troglitazone. AB - Type 2 diabetes has been related to a decrease of mitochondrial DNA (mtDNA) content. In this study, we show increased expression of the peroxisome proliferator-activated receptor-alpha (PPARalpha) and its target genes involved in fatty acid metabolism in skeletal muscle of Zucker Diabetic Fatty (ZDF) (fa/fa) rats. In contrast, the mRNA levels of genes involved in glucose transport and utilization (GLUT4 and phosphofructokinase) were decreased, whereas the expression of pyruvate dehydrogenase kinase 4 (PDK-4), which suppresses glucose oxidation, was increased. The shift from glucose to fatty acids as the source of energy in skeletal muscle of ZDF rats was accompanied by a reduction of subunit 1 of complex I (NADH dehydrogenase subunit 1, ND1) and subunit II of complex IV (cytochrome c oxidase II, COII), two genes of the electronic transport chain encoded by mtDNA. The transcript levels of PPARgamma Coactivator 1 (PGC-1) showed a significant reduction. Treatment with troglitazone (30 mg/kg/day) for 15 days reduced insulin values and reversed the increase in PDK-4 mRNA levels, suggesting improved insulin sensitivity. In addition, troglitazone treatment restored ND1 and PGC-1 expression in skeletal muscle. These results suggest that troglitazone may avoid mitochondrial metabolic derangement during the development of diabetes mellitus 2 in skeletal muscle. PMID- 14563826 TI - Fasting acylation-stimulating protein is predictive of postprandial triglyceride clearance. AB - Postprandial plasma triglyceride (ppTG) and NEFA clearance were stratified by plasma acylation-stimulating protein (ASP) and gender to determine the contribution of fasting ASP in a normal population (70 men; 71 women). In the highest ASP tertile only, ASP decreased over 8 h (90 +/- 9.7 nM to 70 +/- 5.9 nM, P<0.05 males; 61.9 +/- 4.0 nM to 45.6 +/- 6.2 nM, P<0.01 females). Fasting ASP correlated positively with ppTG response. ppTG (P<0.0001, 2-way ANOVA, both genders) and NEFA levels progressively increased from lowest to highest ASP tertile, with the greatest differences in males. By stepwise multiple regression, the best prediction of ppTG was: (fasting ASP + apolipoprotein B + insulin + TG; r=0.806) for men and (fasting ASP + total cholesterol; r=0.574) for women. Leptin, body mass index, and other fasting variables did not improve the prediction. Thus, in men and women, ASP significantly predicted ppTG and NEFA clearance and, based on lower ASP, women may be more ASP sensitive than men. Plasma ASP may be useful as a fasting variable that will provide additional information regarding ppTG and NEFA clearance. PMID- 14563827 TI - Apolipoprotein C-III, metabolic syndrome, and risk of coronary artery disease. AB - Apolipoprotein C-III (apoC-III) is a marker of triglyceride (TG)-rich lipoproteins, which are often increased in metabolic syndrome (MS). The T-455C polymorphism in the insulin-responsive element of the APOC3 gene influences TG and apoC-III levels. To evaluate the contribution of apoC-III levels and T-455C polymorphisms in the coronary artery disease (CAD) risk of MS patients, we studied 873 patients, 549 with CAD and 251 with normal coronary arteries. Patients were classified also as having or not having MS (MS, n = 270; MS-free, n = 603). Lipids, insulin, apolipoprotein levels, and APOC3 T-455C genotypes were evaluated. ApoC-III levels were significantly increased in MS patients, and the probability of having MS was correlated with increasing quartiles of apoC-III levels. MS patients with CAD had significantly higher apoC-III levels than did CAD-free MS patients. The carriership for the -455C variant multiplied the probability of CAD in MS in an allele-specific way and was associated with increased apoC-III and TG levels. Obesity was less frequent in MS carriers of the -455C allele than in MS noncarriers (21.6% vs. 34.8%, P < 0.05). In conclusion, apoC-III-rich lipoprotein metabolism and the APOC3 polymorphism have relevant impacts on the CAD risk of MS patents. PMID- 14563828 TI - Differentially expressed genes in visceral or subcutaneous adipose tissue of obese men and women. AB - There is growing evidence that the distribution of adipose tissue in the body is of importance in the development of metabolic complications of obesity, such as diabetes, hypertension, and hyperlipidemia. The aim of this study was to identify differentially expressed genes in subcutaneous and omental human adipose tissue in obese men, using a subtractive hybridization strategy. From the obtained set of differentially expressed transcripts, we also aimed to identify genes that have a sex-specific pattern of expression in omental or subcutaneous adipose tissue. Representational difference analysis (RDA) was performed on cDNA from subcutaneous and omental fat tissue from a man with extreme abdominal obesity. Forty-four putatively differentially expressed genes were identified. The obtained RDA products were spotted onto glass slides to screen for differential expression in other obese patients by using a microarray hybridization procedure. Five genes were confirmed to be differentially expressed in subcutaneous or omental adipose tissue from male or female obese patients. One gene was detected only in males and was found to be upregulated in subcutaneous tissue. The findings extend previous knowledge that different fat depots have differential gene expression and indicate that sex differences exist in adipose gene expression patterns. PMID- 14563829 TI - ApoB-containing lipoproteins in apoE-deficient mice are not metabolized by the class B scavenger receptor BI. AB - The scavenger receptor class B type I (SR-BI) recognizes a broad variety of lipoprotein ligands, including HDL, LDL, and oxidized LDL. In this study, we investigated whether SR-BI plays a role in the metabolism of cholesterol-rich lipoprotein remnants that accumulate in apolipoprotein E (apoE)(-/-) mice. These particles have an unusual apolipoprotein composition compared with conventional VLDL and LDL, containing mostly apoB-48 as well as substantial amounts of apoA-I and apoA-IV. To study SR-BI activity in vivo, the receptor was overexpressed in apoE(-/-) mice by adenoviral vector-mediated gene transfer. An approximately 10 fold increase in liver SR-BI expression resulted in no detectable alterations in VLDL-sized particles and a modest depletion of cholesterol in intermediate density lipoprotein/LDL-sized lipoprotein particles. This decrease was not attributable to altered secretion of apoB-containing lipoproteins in SR-BI overexpressing mice. To directly assess whether SR-BI metabolizes apoE(-/-) mouse lipoprotein remnants, in vitro assays were performed in both CHO cells and primary hepatocytes expressing high levels of SR-BI. This analysis showed a remarkable deficiency of these particles to serve as substrates for selective lipid uptake, despite high-affinity, high-capacity binding to SR-BI. Taken together, these data establish that SR-BI does not play a direct role in the metabolism of apoE(-/-) mouse lipoprotein remnants. PMID- 14563830 TI - Distinct roles of endoplasmic reticulum cytochrome b5 and fused cytochrome b5 like domain for rat Delta6-desaturase activity. AB - The Delta6-desaturase catalyzes key steps in long-chain polyunsaturated fatty acid biosynthesis. Although the gene coding for this enzyme has been isolated in diverse animal species, the protein structure remains poorly characterized. In this work, rat Delta6-desaturase expressed in COS-7 cells was shown to localize in the endoplasmic reticulum. As the enzyme contains an N-terminal cytochrome b5 like domain, we investigated by site-directed mutagenesis the role of this domain in the enzyme activity. The typical HPGG motif of the cytochrome b5-like domain, and particularly histidine in this motif, is required for the activity of the enzyme, whatever the substrate. Neither endogenous COS-7 cytochrome b5 nor coexpressed rat endoplasmic reticulum cytochrome b5 could rescue the activity of mutated forms of Delta6-desaturase. Moreover, when rat endoplasmic reticulum cytochrome b5 was coexpressed with wild-type desaturase, both proteins interacted and Delta6-desaturase activity was significantly increased. The identified interaction between these proteins is not dependent on the desaturase HPGG motif. These data suggest distinct and essential roles for both the desaturase cytochrome b5-like domain and free endoplasmic reticulum cytochrome b5 for Delta6 desaturase activity. PMID- 14563831 TI - Dietary alpha-linolenic acid reduces COX-2 expression and induces apoptosis of hepatoma cells. AB - Fatty acid synthetase (FAS) is overexpressed in various tumor tissues, and its inhibition and/or malonyl-CoA accumulation have been correlated to apoptosis of tumor cells. It is widely recognized that both omega-3 and omega-6 polyunsaturated fatty acids (PUFAs) depress FAS expression in liver, although epidemiological and experimental reports attribute antitumor properties only to omega-3 PUFA. Therefore, we investigated whether lipogenic gene expression in tumor cells is differently regulated by omega-6 and omega-3 PUFAs. Morris hepatoma 3924A cells were implanted subcutaneously in the hind legs of ACI/T rats preconditioned with high-lipid diets enriched with linoleic acid or alpha linolenic acid. Both-high lipid diets depressed the expression of FAS and acetyl CoA carboxylase in tumor tissue, this effect correlating with a decrease in the mRNA level of their common sterol regulatory element binding protein-1 transcription factor. Hepatoma cells grown in rats on either diet did not accumulate malonyl-CoA. Apoptosis of hepatoma cells was induced by the alpha linolenic acid-enriched diet but not by the linoleic acid-enriched diet. Therefore, in this experimental model, apoptosis is apparently independent of the inhibition of fatty acid synthesis and of malonyl-CoA cytotoxicity. Conversely, it was observed that apoptosis induced by the alpha-linolenic acid-enriched diet correlated with a decrease in arachidonate content in hepatoma cells and decreased cyclooxygenase-2 expression. PMID- 14563832 TI - Smooth muscle raft-like membranes. AB - We developed a method for extracting raft-like, liquid-ordered membranes from the particulate fraction prepared from porcine trachealis smooth muscle. This fraction, which contains most of the plasma membrane in this tissue, was homogenized in the presence of cold 0.5% Triton X-100. After centrifugation, membranes containing high contents of sphingomyelin (SM) and cholesterol and low phosphatidylcholine (PC) contents remained in the pellet. Thirty-five millimolar octyl glucoside (OG) extracted 75% of these membranes from the Triton X-100 resistant pellet. These membranes had low buoyant densities and accounted for 28% of the particulate fraction lipid. Their lipid composition, 22% SM, 60% cholesterol, 11% phosphatidylethanolamine, 8% PC, <1% phosphatidylinositol, and coisolation with 5'-nucleotidase and caveolin-1 suggest that they are liquid ordered membranes. We compared characteristics of OG and Triton X-100 extractions of the particulate fraction. In contrast to Triton X-100 extractions, membranes released from the particulate fraction by OG were mainly collected in low buoyant fractions at densities ranging from 1.05 to 1.11 g/ml and had phospholipid and cholesterol contents consistent with a mixture of liquid-ordered and liquid disordered membranes. Thus, OG extraction of apparent liquid-ordered membranes from Triton X-100-resistant pellets was not due to selective extraction of these membranes. Low buoyant density appears not to be unique for liquid-ordered membranes. PMID- 14563833 TI - Symbiotic N2 fixation activity in relation to C economy of Pisum sativum L. as a function of plant phenology. AB - The relationships between symbiotic nitrogen fixation (SNF) activity and C fluxes were investigated in pea plants (Pisum sativum L. cv. Baccara) using simultaneous 13C and 15N labelling. Analysis of the dynamics of labelled CO2 efflux from the nodulated roots allowed the different components associated with SNF activity to be calculated, together with root and nodule synthetic and maintenance processes. The carbon costs for the synthesis of roots and nodules were similar and decreased with time. Carbon lost by turnover, associated with maintenance processes, decreased with time for nodules while it increased in the roots. Nodule turnover remained higher than root turnover until flowering. The effect of the N source on SNF was investigated using plants supplied with nitrate or plants only fixing N2. SNF per unit nodule biomass (nodule specific activity) was linearly related to the amount of carbon allocated to the nodulated roots regardless of the N source, with regression slopes decreasing across the growth cycle. These regression slopes permitted potential values of SNF specific activity to be defined. SNF activity decreased as the plants aged, presumably because of the combined effects of both increasing C costs of SNF (from 4.0 to 6.7 g C g-1 N) and the limitation of C supply to the nodules. SNF activity competed for C against synthesis and maintenance processes within the nodulated roots. Synthesis was the main limiting factor of SNF, but its importance decreased as the plant aged. At seed-filling, SNF was probably more limited by nodule age than by C supply to the nodulated roots. PMID- 14563834 TI - Isolation and characterization of a maintenance DNA-methyltransferase gene from peach (Prunus persica [L.] Batsch): transcript localization in vegetative and reproductive meristems of triple buds. AB - A cDNA coding for a DNA (cytosine-5)-methyltransferase (METase) was isolated from peach (Prunus persica [L.] Batsch) and the corresponding gene designated as PpMETI. The latter encoded a predicted polypeptide of 1564 amino acid residues and harboured all the functional domains conserved in the maintenance METases group type I. PpMETI was a single copy in the cultivar Chiripa which was used as a model in the present study. Expression analyses revealed that PpMETI transcripts were more abundant in tissues with actively proliferating cells such as apical tips, uncurled leaves, elongating herbaceous stems, and small immature fruits. Peach plants bear bud clusters (triads or triple buds), consisting of two lateral and one central bud with floral and vegetative fates, respectively. PpMETI in situ hybridization was performed in triple buds during their entire developmental cycle. High and low levels of PpMETI transcript were related to burst and quiescence of vegetative growth, respectively. Message localization distinguished lateral from central buds during the meristem switch to the floral phase. In fact, the PpMETI message was abundant in the L1 layer of protruding domes, a morphological trait marking the beginning of floral transition. The PpMETI transcript was also monitored during organ flower formation. Altogether, these data suggest a relationship between DNA replication and PpMETI gene expression. PMID- 14563835 TI - Mutation in the ap2-6 allele causes recognition of a cryptic splice site. AB - Mutations in the homeotic gene APETALA2 of Arabidopsis thaliana cause severe developmental alterations, most prominently homeotic floral organ replacements from petals to carpels and petals to stamens in the outer two floral whorls. To date, ten different alleles have been identified conferring phenotypes of various degrees. Of these ten alleles, only three have been characterized at the sequence level. The identification of the sequence alteration in the ap2-6 allele is reported here. In ap2-6 a single G.C to A.T transition occurred at the 3' end of intron 6 (position 1342) which leads to a dinucleotide loss at the mRNA level. This change is consistent with the G.C to A.T transition destroying a conserved dinucleotide motif (AG) required for proper splice recognition and with the resulting recognition of the next available downstream AG dinucleotide which in AP2 is immediately adjacent to the authentic 3' splice site. The dinucleotide loss will cause a frameshift, the translation of three incorrect amino acids and a premature stop codon resulting in a truncation of the AP2 sequence within the AP2-R2 domain. Such a truncation is predicted to impact severely on the function of AP2 and is consistent with the observed phenotype. PMID- 14563836 TI - Caution! DNA crossing: crystal structures of Holliday junctions. PMID- 14563837 TI - Phosphorylation by glycogen synthase kinase-3 controls c-myc proteolysis and subnuclear localization. AB - The c-Myc protein is a transcription factor that is a central regulator of cell growth and proliferation. Thr-58 is a major phosphorylation site in c-Myc and is a mutational hotspot in Burkitt's and other aggressive human lymphomas, indicating that Thr-58 phosphorylation restricts the oncogenic potential of c Myc. Mutation of Thr-58 is also associated with increased c-Myc protein stability. Here we show that inhibition of glycogen synthase kinase-3 (GSK-3) activity with lithium increases c-Myc stability and inhibits phosphorylation of c Myc specifically at Thr-58 in vivo. Conversely, overexpression of GSK-3 alpha or GSK-3 beta enhances Thr-58 phosphorylation and ubiquitination of c-Myc. Together, these observations suggest that phosphorylation of Thr-58 mediated by GSK-3 facilitates c-Myc rapid proteolysis by the ubiquitin pathway. Furthermore, we demonstrate that GSK-3 binds c-Myc in vivo and in vitro and that GSK-3 colocalizes with c-Myc in the nucleus, strongly arguing that GSK-3 is the c-Myc Thr-58 kinase. We found that c-MycS, which lacks the N-terminal 100 amino acids of c-Myc, is unable to bind GSK-3; however, mutation of Ser-62, the priming phosphorylation site necessary for Thr-58 phosphorylation, does not disrupt GSK-3 binding. Finally, we show that Thr-58 phosphorylation alters the subnuclear localization of c-Myc, enhancing its localization to discrete nuclear bodies together with GSK-3. PMID- 14563838 TI - Activation of the canonical beta-catenin pathway by histamine. AB - Histamine signaling is a principal regulator in a variety of pathophysiological processes including inflammation, gastric acid secretion, neurotransmission, and tumor growth. We report that histamine stimulation causes transactivation of a T cell factor/beta-catenin-responsive construct in HeLa cells and in the SW-480 colon cell line, whereas histamine did not effect transactivation of a construct containing the mutated response construct FOP. On the protein level, histamine treatment increases phosphorylation of glycogen synthase kinase 3-beta in HeLa cells, murine macrophages, and DLD-1, HT-29, and SW-480 colon cell lines. Furthermore, histamine also decreases the phosphorylated beta-catenin content in HeLa cells and murine macrophages. Finally, pharmacological inhibitors of the histamine H1 receptor counteracted histamine-induced T cell factor/beta-catenin responsive construct transactivation and the dephosphorylation of beta-catenin in HeLa cells and in macrophages. We conclude that the canonical beta-catenin pathway acts downstream of the histamine receptor H1 in a variety of cell types. The observation that inflammatory molecules, like histamine, activate the beta catenin pathway may provide a molecular explanation for a possible link between inflammation and cancer. PMID- 14563839 TI - Non-canonical eukaryotic glutaminyl- and glutamyl-tRNA synthetases form mitochondrial aminoacyl-tRNA in Trypanosoma brucei. AB - Glutaminyl-tRNA synthetase is thought to be absent from organelles. Instead, Gln tRNA is formed via the transamidation pathway, the other route to this essential compound in protein biosynthesis. However, it was previously shown that glutaminyl-tRNA synthetase activity is present in Leishmania mitochondria. This work identifies genes encoding glutaminyl- and glutamyl-tRNA synthetase in the closely related organism Trypanosoma brucei. Down-regulation of their respective gene products by RNA interference showed that (i) they are essential for the growth of insect stage T. brucei and (ii) they are responsible for essentially all of the glutaminyl- and glutamyl-tRNA synthetase activity detected in both the cytosol and the mitochondria. In vitro aminoacylation experiments with the recombinant T. brucei enzymes and total tRNA confirmed the identity of the two aminoacyl-tRNA synthetases. Interestingly, T. brucei uses the same eukaryotic type glutaminyl-tRNA synthetase to form mitochondrial and cytosolic Gln-tRNA. The formation of Glu-tRNA in mitochondria and the cytoplasm is catalyzed by a single eukaryotic-type discriminating glutamyl-tRNA synthetase. T. brucei, similar to Leishmania, imports all of its mitochondrial tRNAs from the cytosol. The use of these two eukaryotic-type enzymes in mitochondria may therefore reflect an adaptation to the situation in which the cytosol and mitochondria use the same set of tRNAs. PMID- 14563841 TI - Modulation of hyperthermophilic DNA polymerase activity by archaeal chromatin proteins. AB - Sulfolobus synthesizes a large quantity of highly conserved 7-kDa DNA-binding proteins suspected to be involved in chromosomal organization. The effect of the 7-kDa proteins on the polymerization and 3'-5' exonuclease activities of a family B DNA polymerase (polB1) from the hyperthermophilic archaeon Sulfolobus solfataricus was investigated. polB1 degraded both single-stranded DNA and double stranded DNA at similar rates in vitro at temperatures of physiological relevance. The 7-kDa proteins were capable of significantly inhibiting the excision and enhancing the extension of matched template primers by the polymerase. However, the proteins did not protect single-stranded DNA from cleavage by polB1. In addition, the 7-kDa proteins did not affect the proofreading ability of polB1 and were not inhibitory to the excision of mismatched primers by the polymerase. The dNTP concentrations required for the effective inhibition of the 3'-5' exonuclease activity of polB1 were lowered from approximately 1 mm in the absence of the 7-kDa proteins to approximately 50 microm in the presence of the proteins at 65 degrees C. Our data suggest that the 7-kDa chromatin proteins serve to modulate the extension and excision activities of the hyperthermophilic DNA polymerase, reducing the cost of proofreading by the enzyme at high temperature. PMID- 14563840 TI - Insig-dependent ubiquitination and degradation of mammalian 3-hydroxy-3 methylglutaryl-CoA reductase stimulated by sterols and geranylgeraniol. AB - The endoplasmic reticulum enzyme 3-hydroxy-3-methylglutaryl-CoA reductase produces mevalonate, which is converted to sterols and to other products, including geranylgeraniol groups attached to proteins. The enzyme is known to be ubiquitinated and rapidly degraded when sterols and nonsterol end products of mevalonate metabolism accumulate in cells. Here, we use RNA interference to show that sterol-accelerated ubiquitination of reductase requires Insig-1 and Insig-2, membrane-bound proteins of the endoplasmic reticulum that were shown previously to accelerate degradation of reductase when overexpressed by transfection. Alanine substitution experiments reveal that binding of reductase to Insigs and subsequent ubiquitination require the tetrapeptide sequence YIYF in the second membrane-spanning helix of reductase. The YIYF peptide is also found in the sterol-sensing domain of SCAP, another protein that binds to Insigs in a sterol stimulated fashion. When lysine 248 of reductase is substituted with arginine, Insig binding persists, but the reductase is no longer ubiquitinated and degradation is markedly slowed. Lysine 248 is predicted to lie immediately adjacent to a membrane-spanning helix, suggesting that a membrane-bound ubiquitin transferase is responsible. Finally, we show that Insig-dependent, sterol stimulated degradation of reductase is further accelerated when cells are also supplied with the 20-carbon isoprenoid geranylgeraniol, but not the 15-carbon farnesol, raising the possibility that the nonsterol potentiator of reductase regulation is a geranylgeranylated protein. PMID- 14563842 TI - The D246V mutant of DNA polymerase beta misincorporates nucleotides: evidence for a role for the flexible loop in DNA positioning within the active site. AB - DNA polymerase beta, a member of the X family of DNA polymerases, is known to be involved in base excision repair. A key to determining the biochemical properties of this DNA polymerase is structure-function studies of site-specific mutants that result in substitution of particular amino acids at critical sites. In a previous genetic screen, we identified three 3'-azido-2',3'-dideoxythymidine 5' triphosphate-resistant mutants, namely E249K, D246V, and R253M, of polymerase beta in the flexible loop of the palm domain. In this work, we perform an extensive kinetic analysis to investigate the role of the D246V mutant on polymerase fidelity. We find that D246V misincorporates T opposite template bases G and C. The mechanistic basis of misincorporation appears to be altered DNA positioning within the active site. We provide evidence that the fidelity of D246V is greatly affected by the base that is 5' of the templating base. We propose that the Asp residue at position 246 helps to maintain the proper positioning of the DNA within the polymerase active site and maintains the fidelity of polymerase beta. Altogether, the results suggest that the flexible loop domain of polymerase beta plays a major role in its fidelity. PMID- 14563843 TI - Characterization of the interaction of the stress kinase SPAK with the Na+-K+-2Cl cotransporter in the nervous system: evidence for a scaffolding role of the kinase. AB - Activity of heterologously expressed NKCC1 was analyzed under basal and activated conditions in the presence and absence of binding of Ste20-related proline alanine-rich kinase (SPAK). Mutant NKCC1 that lacks the ability to bind to this kinase showed K+ transport function identical to wild-type NKCC1. Thus, preventing the binding of the kinase to the cotransporter does not affect cotransporter function. In contrast, several experiments suggest a possible role for SPAK as a scaffolding protein. First, Western blot analysis revealed the presence, and in some tissues abundance, of truncated forms of SPAK and OSR1 in which the kinase domains are affected and thus lack kinase activity. Second, a yeast two-hybrid screen of proteins that interact with the regulatory (binding) domain of SPAK identified several proteins all involved in cellular stress pathways. Third, p38, one of the three major MAPKs, can be coimmunoprecipitated with SPAK and with NKCC1 in an activity-dependent manner. The amount of p38 coimmunoprecipitated with the kinase and the cotransporter significantly decreases upon cellular stress, whereas the interaction of the kinase with NKCC1 remains unchanged. These findings suggest that cation-chloride cotransporters might act as "sensors" for cellular stress, and SPAK, by interacting with the cotransporter, serves as an intermediate in the response to cellular stress. PMID- 14563844 TI - Initial binding of the broad spectrum antiviral nucleoside ribavirin to the hepatitis C virus RNA polymerase. AB - Ribavirin is a broad spectrum antiviral nucleoside that displays activity against a variety of RNA and DNA viruses. Ribavirin is currently used in combination with interferon-alpha for the treatment of hepatitis C virus (HCV) infection and was recently shown to be directly incorporated by the HCV RNA polymerase into RNA products. This capacity ultimately leads to increased mutation rates and drastically reduces the viral fitness. As a first step toward elucidating the nature of the specific interaction between ribavirin and the HCV polymerase, we have utilized fluorescence spectroscopy to monitor precisely the binding of ribavirin triphosphate (RTP) to the viral polymerase. This spectroscopic approach allowed us to clearly separate the RTP binding activity from the concomitant catalytic steps. We report here the first detailed study of the binding kinetics and thermodynamic parameters involved in the interaction between RTP and an RNA polymerase. We demonstrate that RTP binds to the same active site as nucleotides. Furthermore, we provide evidence that the HCV polymerase cannot only bind to RTP but also to nonphosphorylated ribavirin, albeit with less affinity. By using various combinations of template-primers, we also demonstrate that base pairing is not involved in the initial binding of RTP to the HCV polymerase. Based on the results of circular dichroism and denaturation studies, we show that the RNA polymerase undergoes subtle conformational changes upon the binding of RTP, although the interaction does not significantly modify the stability of the protein. Finally, although metal ions are required for catalytic activity, they are not required for the initial binding of RTP to the polymerase. Such quantitative analyses are of primary importance for the rational design of new ribavirin analogues of potential therapeutic value and provide crucial insights on the interaction between RTP and the HCV RNA polymerase. PMID- 14563845 TI - Tumor necrosis factor-alpha activates NFkappaB to inhibit renin transcription by targeting cAMP-responsive element. AB - Tumor necrosis factor-alpha (TNFalpha) is known to inhibit renin gene expression in juxtaglomerular cells, which are the main source of renin in vivo. In the present study we aimed to characterize the intracellular mechanisms of TNFalpha signaling to renin gene in the mouse juxtaglomerular cell line As4.1. TNFalpha was found to activate NFkappaB, which is one of the principal intracellular mediators of TNFalpha signal transduction. Constitutive activation of NFkappaB suppressed renin gene transcription, but NFkappaB appeared not to target the NFkappaB binding sites in the renin promoter. Thus, NFkappaB, but not the canonical NFkappaB binding sequences in the renin promoter, seemed to be involved in the suppression of renin transcription by TNFalpha. Deletion/mutation analysis revealed that the effect of TNFalpha on renin gene is transmitted by a cAMP responsive element (CRE) located at -2697 to -2690. Mobility shift/supershift assays evidenced for the presence of NFkappaB proteins in the complex that binds to mouse renin CRE. Our results strongly suggest that NFkappaB mediates the effect of TNFalpha on renin transcription targeting a CRE in the mouse renin promoter. PMID- 14563846 TI - Structure of Plasmodium falciparum triose-phosphate isomerase-2-phosphoglycerate complex at 1.1-A resolution. AB - Triose-phosphate isomerase, a key enzyme of the glycolytic pathway, catalyzes the isomerization of dihydroxy acetone phosphate and glyceraldehyde 3-phosphate. In this communication we report the crystal structure of Plasmodium falciparum triose-phosphate isomerase complexed to the inhibitor 2-phosphoglycerate at 1.1-A resolution. The crystallographic asymmetric unit contains a dimeric molecule. The inhibitor bound to one of the subunits in which the flexible catalytic loop 6 is in the open conformation has been cleaved into two fragments presumably due to radiation damage. The cleavage products have been tentatively identified as 2 oxoglycerate and meta-phosphate. The intact 2-phosphoglycerate bound to the active site of the other subunit has been observed in two different orientations. The active site loop in this subunit is in both open and "closed" conformations, although the open form is predominant. Concomitant with the loop closure, Phe-96, Leu-167, and residues 208-211 (YGGS) are also observed in dual conformations in the B-subunit. Detailed comparison of the active-site geometry in the present case to the Saccharomyces cerevisiae triose-phosphate isomerase-dihydroxy acetone phosphate and Leishmania mexicana triose-phosphate isomerase-phosphoglycolate complexes, which have also been determined at atomic resolution, shows that certain interactions are common to the three structures, although 2 phosphoglycerate is neither a substrate nor a transition state analogue. PMID- 14563847 TI - Targeted gene disruption of glycerol-3-phosphate dehydrogenase in Colletotrichum gloeosporioides reveals evidence that glycerol is a significant transferred nutrient from host plant to fungal pathogen. AB - Unidirectional transfer of nutrients from plant host to pathogen represents a most revealing aspect of the parasitic lifestyle of plant pathogens. Whereas much effort has been focused on sugars and amino acids, the identification of other significant metabolites is equally important for comprehensive characterization of metabolic interactions between plants and biotrophic fungal pathogens. Employing a strategy of targeted gene disruption, we generated a mutant strain (gpdhDelta) defective in glycerol-3-phosphate dehydrogenase in a hemibiotrophic plant pathogen, Colletotrichum gloeosporioides f.sp. malvae. The gpdhDelta strain had severe defects in carbon utilization as it could use neither glucose nor amino acids for sustained growth. Although the mutant mycelia were able to grow on potato dextrose agar medium, they displayed arrhythmicity in growth and failure to conidiate. The metabolic defect of gpdhDelta could be entirely ameliorated by glycerol in chemically defined minimal medium. Furthermore, glycerol was the one and only metabolite that could restore rhythmic growth and conidiation of gpdhDelta. Despite the profound defects in carbon source utilization, in planta the gpdhDelta strain exhibited normal pathogenicity, proceeded normally in its life cycle, and produced abundant conidia. Analysis of plant tissues at the peripheral zone of fungal infection sites revealed a time dependent reduction in glycerol content. This study provides strong evidence for a role of glycerol as a significant transferred metabolite from plant to fungal pathogen. PMID- 14563848 TI - The serine/threonine kinase Nek6 is required for cell cycle progression through mitosis. AB - The Aspergillus nidulans protein NIMA (never in mitosis, gene A) is a protein kinase required for the initiation of mitosis, whereas its inactivation is necessary for mitotic exit. Here, we demonstrate that human NIMA-related kinase 6 (Nek6) is required for mitotic progression of human cells. Nek6 is phosphorylated and activated during M phase. Inhibition of Nek6 function by either overexpression of an inactive Nek6 mutant or elimination of endogenous Nek6 by siRNA arrests cells in M phase and triggers apoptosis. Time-lapse recording of the cell cycle progression of cells expressing kinase-inactive Nek6 reveals mitotic arrest at the metaphase stage prior to cells entering apoptosis. In contrast to NIMA and the closely related mammalian Nek2 kinase, which regulate centrosome function and separation, our data demonstrate an important function for Nek6 during mitosis and suggest that Nek6 kinase is required for metaphase anaphase transition. PMID- 14563849 TI - Molecular determinants of proton modulation of glycine receptors. AB - Extracellular pH regulates glycine receptors through an unknown mechanism. Here we demonstrate that acidic pH remarkably inhibited glycine-activated whole-cell currents in recombinant glycine alpha1 and alpha1beta receptors transiently expressed in human embryonic kidney 293 cells. The proton effect was voltage independent and pharmacologically competed with glycine receptor agonist glycine and antagonist strychnine. Using site-directed mutagenesis, we have identified an N-terminal domain that is essential for proton-induced inhibition of glycine current. In alpha1 homomers, removal of the hydroxyl group by mutation of residue Thr-112 to Ala or Phe abolished inhibition of glycine currents by acidification. In contrast, mutation of Thr-112 to another hydroxylated residue (Tyr) produced receptors that retained partial proton sensitivity. In alpha1beta heteromers, a single mutation of the beta subunit T135A, which is homologous to alpha1 Thr-112, reduced proton sensitivity, whereas the double mutation alpha1(T112A)beta(T135A) almost completely eliminated the proton sensitivity. In addition, the mutation alpha1 H109A greatly reduced sensitivity to protons in homomeric alpha1 receptors. The results demonstrate that extracellular pH can regulate the function of glycine alpha1 and alpha1beta receptors. An extracellular domain consisting of Thr-112 and His-109 at the alpha1 subunit and Thr-135 at the beta subunit plays a critical role in determining proton modulation of glycine receptor function. PMID- 14563850 TI - Tom1, a VHS domain-containing protein, interacts with tollip, ubiquitin, and clathrin. AB - The gene for Tom1 was initially identified as a specific target of the oncogene v myb. The Tom1 protein belongs to the VHS domain-containing protein family, and it has a GAT domain in a central part as well as an N-terminal VHS domain. VHS domain-containing proteins, including Hrs/Vps27, STAM, and GGA proteins, have been implicated in intracellular trafficking and sorting, but the role of Tom1 has not yet been elucidated. In this study, we found that Tom1 binds directly with ubiquitin chains and Tollip, which was initially isolated as a mediator of interleukin-1 signaling and has a capacity to bind ubiquitin chains. Gel filtration and subsequent Western blot analysis showed that endogenous Tom1 associates with Tollip to form a complex. In addition, Tom1 was found to be capable of binding to clathrin heavy chain through a typical clathrin-binding motif. Fluorescence microscopic analysis revealed that green fluorescent protein Tom1 was localized predominantly in the cytoplasm, whereas its mutant with deletion of the clathrin-binding motif had a diffuse localization throughout the cell. Thus, we propose that a Tom1-Tollip complex functions as a factor that links polyubiquitinated proteins to clathrin. PMID- 14563851 TI - Identification of the key amino acids of glial cell line-derived neurotrophic factor family receptor alpha1 involved in its biological function. AB - Glial cell line-derived neurotrophic factor (GDNF) plays a critical role in neurodevelopment and survival of midbrain dopaminergic and spinal motor neurons in vitro and in vivo. The biological actions of GDNF are mediated by a two receptor complex consisting of a glycosylphosphatidylinositol-linked cell surface molecule, the GDNF family receptor alpha1 (GFRalpha1), and receptor protein tyrosine kinase Ret. Although structural analysis of GDNF has been extensively examined, less is known about the structural basis of GFRalpha1 function. In this study, based on evolutionary trace method and relative solvent accessibility prediction of residues, a set of trace residues that are solvent-accessible was selected for site-directed mutagenesis. A series of GFRalpha1 mutations was made, and PC12 cell lines stably expressing different GFRalpha1 mutants were generated. According to the survival and differentiation responses of these stable PC12 cells upon GDNF stimulation and the GDNF-GFRalpha1-Ret interaction assay, residues 152NN153, Arg259, and 316SNS318 in the GFRalpha1 central region were found to be critical for GFRalpha1 binding to GDNF and eliciting downstream signal transduction. The single mutation R259A in the GFRalpha1 molecule simultaneously lost its binding ability to GDNF and Ret. However N152A/N153A or S316A/N317A/S318A mutation in the GFRalpha1 molecule still retained the ability to bind with Ret. These findings suggest that distinct structural elements in GFRalpha1 may be involved in binding to GDNF and Ret. PMID- 14563852 TI - Characterization of the localization and proteolytic activity of the SUMO specific protease, SENP1. AB - Modification of proteins by small ubiquitin-like modifier (SUMO) plays an important role in the function, compartmentalization, and stability of target proteins, contributing to the regulation of diverse processes. SUMO-1 modification can be regulated not only at the level of conjugation; it may also be reversed by a class of proteases known as the SUMO-specific proteases. However, current understanding of the regulation, specificity, and function of these proteases remains limited. In this study, we characterize aspects of the compartmentalization and proteolytic activity of the mammalian SUMO-specific protease, SENP1, providing insight into its function and regulation. We demonstrate the presence of a single nonconsensus nuclear localization signal within the N terminus of the protein, the mutation of which results in pronounced cytoplasmic accumulation in contrast to the nuclear accumulation of the parental protein. In addition, we observe that the N terminus of the protein may be essential for the correct regulation of the protease, since expression of the core domain alone results in limited expression and loss of SUMO-1, indicative of constitutive catalytic activity. Consistent with the prediction that the protease is a member of the cysteine family of proteases, we mutated a key cysteine residue and observed that expression of this catalytic mutant had a dominant negative phenotype, resulting in the accumulation of high molecular weight SUMO-1 conjugates. Furthermore, we demonstrate that SENP1 may itself be a target for SUMO-1 modification occurring at a nonconsensus site. Finally, we demonstrate that SENP1 localization is influenced by expression and localization of SUMO-1 conjugated target proteins within the cell. PMID- 14563853 TI - Evidence for a second interaction between the regulatory amino-terminal and central output domains of the response regulator NtrC (nitrogen regulator I) in Escherichia coli. AB - Nitrogen limitation in Escherichia coli activates about 100 genes. Their expression requires the response regulator NtrC (also called nitrogen regulator I or NR(I)). Phosphorylation of the amino-terminal domain (NTD) of NtrC activates the neighboring central domain and leads to transcriptional activation from promoters that require sigma(54)-containing RNA polymerase. The NTD has five beta strands alternating with five alpha helices. Phosphorylation of aspartate 54 has been shown to reposition alpha helix 3 to beta strand 5 (the "3445 face") within the NTD. To further study the interactions between the amino-terminal and central domains, we isolated strains with alterations in the NTD that were able to grow on a poor nitrogen source in the absence of phosphorylation by the cognate sensor kinase. We identified strains with alterations located in the 3445 face and alpha helix 5. Both types of alterations stimulated central domain activities. The alpha helix 5 alterations differed from those in the 3445 face. They did not cause a large scale conformational change in the NTD, which is not necessary for transcriptional activation in these mutants. Yeast two-hybrid analysis indicated that substitutions in both alpha helix 5 and the 3445 face diminish the interaction between the NTD and the central domain. Our results suggest that alpha helix 5 of the NTD, in addition to the 3445 face, interacts with the central domain. We present a model of interdomain signal transduction that proposes different functions for alpha helix 5 and the 3445 face. PMID- 14563854 TI - Involvement of Cdc42 signaling in apoA-I-induced cholesterol efflux. AB - Cholesterol efflux, an important mechanism by which high density lipoproteins (HDL) protect against atherosclerosis, is initiated by docking of apolipoprotein A-I (apoA-I), a major HDL protein, to specific binding sites followed by activation of ATP-binding cassette transporter A1 (ABCA1) and translocation of cholesterol from intracellular compartments to the exofacial monolayer of the plasma membrane where it is accessible to HDL. In this report, we investigated potential signal transduction pathways that may link apoA-I binding to cholesterol translocation to the plasma membrane and cholesterol efflux. By using pull-down assays we found that apoA-I substantially increased the amount of activated Cdc42, Rac1, and Rho in human fibroblasts. Moreover, apoA-I induced actin polymerization, which is known to be controlled by Rho family G proteins. Inhibition of Cdc42 and Rac1 with Clostridium difficile toxin B inhibited apoA-I induced cholesterol efflux, whereas inhibition of Rho with Clostridium botulinum C3-exoenzyme exerted opposite effects. Adenoviral expression of a Cdc42(T17N) dominant negative mutant substantially reduced apoA-I-induced cholesterol efflux, whereas dominant negative Rac1(T17N) had no effect. We further found that two downstream effectors of Cdc42/Rac1 signaling, c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38 MAPK), are activated by apoA-I. Pharmacological inhibition of JNK but not p38 MAPK decreased apoA-I-induced cholesterol efflux, whereas anisomycin and hydrogen peroxide, two direct JNK activators, could partially substitute for apoA-I in its ability to induce cholesterol efflux. These results for the first time demonstrate activation of Rho family G proteins and stress kinases by apoA-I and implicate the involvement of Cdc42 and JNK in the apoA-I-induced cholesterol efflux. PMID- 14563856 TI - Putative interhelical interactions within the PheP protein revealed by second site suppressor analysis. AB - Highly conserved glycine residues within span I and span II of the phenylalanine and tyrosine transporter PheP were shown to be important for the function of the wild-type protein. Replacement by amino acids with increasing side chain volume led to progressive loss of transport activity. Second-site suppression studies performed with a number of the primary mutants revealed a tight packing arrangement between spans I and II that is important for function and an additional interaction between spans I and III. We also postulate that a third motif, GXXIG, present in span I and highly conserved within different members of the amino acid-polyamine-organocation family, may function as a dimerization motif. Surprisingly, other highly conserved residues, such as Y60 and L41, could be replaced by various residues with no apparent loss of activity. PMID- 14563855 TI - The human herpes virus 8-encoded viral FLICE-inhibitory protein induces cellular transformation via NF-kappaB activation. AB - Infection with human herpes virus 8 (HHV8) has been associated with Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman's disease. HHV8 encodes for a viral FLICE-inhibitory protein (vFLIP), designated K13, which resembles the prodomain of caspase-8 in structure and has been shown to protect cells against death receptor-induced apoptosis in vitro and in vivo. In this report, we present evidence that HHV8 vFLIP also possesses the unique ability of transforming Rat-1 and Balb/3T3 fibroblast cells, which is not shared by other vFLIPs. Rat-1 cells expressing HHV8 vFLIP form colonies in soft agar and form tumors in nude mice. The transforming ability of HHV8 vFLIP is associated with the activation of the NF-kappaB pathway and is blocked by molecular and chemical inhibitors of this pathway. Our results suggest that vFLIP K13 has activity beyond its role as an inhibitor of death receptor signaling and may play a causative role in the pathogenesis of HHV8-associated malignancies. Furthermore, inhibitors of the NF-kappaB pathway may have a role in the treatment of malignancies linked to HHV8 infection. PMID- 14563857 TI - Identification and characterization of the emhABC efflux system for polycyclic aromatic hydrocarbons in Pseudomonas fluorescens cLP6a. AB - The hydrocarbon-degrading environmental isolate Pseudomonas fluorescens LP6a possesses an active efflux mechanism for the polycyclic aromatic hydrocarbons phenanthrene, anthracene, and fluoranthene but not for naphthalene or toluene. PCR was used to detect efflux pump genes belonging to the resistance-nodulation cell division (RND) superfamily in a plasmid-cured derivative, P. fluorescens cLP6a, which is unable to metabolize hydrocarbons. One RND pump, whose gene was identified in P. fluorescens cLP6a and was designated emhB, showed homology to the multidrug and solvent efflux pumps in Pseudomonas aeruginosa and Pseudomonas putida. The emhB gene is located in a gene cluster with the emhA and emhC genes, which encode the membrane fusion protein and outer membrane protein components of the efflux system, respectively. Disruption of emhB by insertion of an antibiotic resistance cassette demonstrated that the corresponding gene product was responsible for the efflux of polycyclic aromatic hydrocarbons. The emhB gene disruption did not affect the resistance of P. fluorescens cLP6a to tetracycline, erythromycin, trimethoprim, or streptomycin, but it did decrease resistance to chloramphenicol and nalidixic acid, indicating that the EmhABC system also functions in the efflux of these compounds and has an unusual selectivity. Phenanthrene efflux was observed in P. aeruginosa, P. putida, and Burkholderia cepacia but not in Azotobacter vinelandii. Polycyclic aromatic hydrocarbons represent a new class of nontoxic, highly hydrophobic compounds that are substrates of RND efflux systems, and the EmhABC system in P. fluorescens cLP6a has a narrow substrate range for these hydrocarbons and certain antibiotics. PMID- 14563858 TI - Involvement of an inducible fructose phosphotransferase operon in Streptococcus gordonii biofilm formation. AB - Oral streptococci, such as Streptococcus gordonii, are the predominant early colonizers that initiate biofilm formation on tooth surfaces. Investigation of an S. gordonii::Tn917-lac biofilm-defective mutant isolated by using an in vitro biofilm formation assay showed that the transposon insertion is near the 3' end of an open reading frame (ORF) encoding a protein homologous to Streptococcus mutans FruK. Three genes, fruR, fruK, and fruI, were predicted to encode polypeptides that are part of the fructose phosphotransferase system (PTS) in S. gordonii. These proteins, FruR, FruK, and FruI, are homologous to proteins encoded by the inducible fruRKI operon of S. mutans. In S. mutans, FruR is a transcriptional repressor, FruK is a fructose-1-phosphate kinase, and FruI is the fructose-specific enzyme II (fructose permease) of the phosphoenolpyruvate dependent sugar PTS. Reverse transcription-PCR confirmed that fruR, fruK, and fruI are cotranscribed as an operon in S. gordonii, and the transposon insertion in S. gordonii fruK::Tn917-lac resulted in a nonpolar mutation. Nonpolar inactivation of either fruK or fruI generated by allelic replacement resulted in a biofilm-defective phenotype, whereas a nonpolar mutant with an inactivated fruR gene retained the ability to form a biofilm. Expression of fruK, as measured by the beta-galactosidase activity of the fruK::Tn917-lac mutant, was observed to be growth phase dependent and was enhanced when the mutant was grown in media with high levels of fructose, sucrose, xylitol, and human serum, indicating that the fructose PTS operon was fructose and xylitol inducible, similar to the S. mutans fructose PTS. The induction by fructose was inhibited by the presence of glucose, indicating that glucose is able to catabolite repress fruK expression. Nonpolar inactivation of the fruR gene in the fruK::Tn917-lac mutant resulted in a greater increase in beta-galactosidase activity when the organism was grown in media supplemented with fructose, confirming that fruR is a transcriptional repressor of the fructose PTS operon. These results suggest that the regulation of fructose transport and metabolism in S. gordonii is intricately tied to carbon catabolite control and the ability to form biofilms. Carbon catabolite control, which modulates carbon flux in response to environmental nutritional levels, appears to be important in the regulation of bacterial biofilms. PMID- 14563859 TI - Morphogenesis of bacillus spore surfaces. AB - Spores produced by bacilli are encased in a proteinaceous multilayered coat and, in some species (including Bacillus anthracis), further surrounded by a glycoprotein-containing exosporium. To characterize bacillus spore surface morphology and to identify proteins that direct formation of coat surface features, we used atomic-force microscopy (AFM) to image the surfaces of wild type and mutant spores of Bacillus subtilis, as well as the spore surfaces of Bacillus cereus 569 and the Sterne strain of Bacillus anthracis. This analysis revealed that the coat surfaces in these strains are populated by a series of bumps ranging between 7 and 40 nm in diameter, depending on the species. Furthermore, a series of ridges encircled the spore, most of which were oriented along the long axis of the spore. The structures of these ridges differ sufficiently between species to permit species-specific identification. We propose that ridges are formed early in spore formation, when the spore volume likely decreases, and that when the spore swells during germination the ridges unfold. AFM analysis of a set of B. subtilis coat protein gene mutants revealed three coat proteins with roles in coat surface morphology: CotA, CotB, and CotE. Our data indicate novel roles for CotA and CotB in ridge pattern formation. Taken together, these results are consistent with the view that the coat is not inert. Rather, the coat is a dynamic structure that accommodates changes in spore volume. PMID- 14563860 TI - Sequence diversity of Treponema pallidum subsp. pallidum tprK in human syphilis lesions and rabbit-propagated isolates. AB - The tprK gene of Treponema pallidum subsp. pallidum, the causative agent of venereal syphilis, belongs to a 12-member gene family and encodes a protein with a predicted cleavable signal sequence and predicted transmembrane domains. Except for the Nichols type strain, all rabbit-propagated isolates of T. pallidum examined thus far are comprised of mixed populations of organisms with heterogeneous tprK sequences. We show that tprK sequences in treponemes obtained directly from syphilis patients are also heterogeneous. Clustering analysis demonstrates that primary chancre tprK sequences are more likely to cluster within a sample than among samples and that tighter clustering is seen within chancre samples than within rabbit-propagated isolates. Closer analysis of tprK sequences from a rabbit-propagated isolate reveals that individual variable regions have different levels of diversity, suggesting that variable regions may have different intrinsic rates of sequence change or may be under different levels of selection. Most variable regions show increased sequence diversity upon passage. We speculate that the diversification of tprK during infection allows organisms to evade the host immune response, contributing to reinfection and persistent infection. PMID- 14563861 TI - Molecular evolution of the dotA gene in Legionella pneumophila. AB - The molecular evolution of dotA, which is related to the virulence of Legionella pneumophila, was investigated by comparing the sequences of 15 reference strains (serogroups 1 to 15). It was found that dotA has a complex mosaic structure. The whole dotA gene of Legionella pneumophila subsp. pneumophila serogroups 2, 6, and 12 has been transferred from Legionella pneumophila subsp. fraseri. A discrepancy was found between the trees inferred from the nucleotide and deduced amino acid sequences of dotA, which suggests that multiple hits, resulting in synonymous substitutions, have occurred. Gene phylogenies inferred from three different segments (the 5'-end region, the central, large periplasmic domain, and the 3' end region) showed impressively dissimilar topologies. This was concordant with the sequence polymorphisms, indicating that each region has experienced an independent evolutionary history, and was evident even within the same domain of each strain. For example, the PP2 domain was found to have a heterogeneous structure, which led us hypothesize that the dotA gene of L. pneumophila may have originated from two or more different sources. Comparisons of synonymous and nonsynonymous substitutions demonstrated that the PP2 domain has been under strong selective pressure with respect to amino acid change. Split decomposition analysis also supported the intragenic recombination of dotA. Multiple recombinational exchange within the dotA gene, encoding an integral cytoplasmic membrane protein that is secreted, probably provided increased fitness in certain environmental niches, such as within a particular biofilm community or species of amoebae. PMID- 14563862 TI - Molecular architecture of the regulatory Locus sae of Staphylococcus aureus and its impact on expression of virulence factors. AB - We characterized the sae operon, a global regulator for virulence gene expression in Staphylococcus aureus. A Tn917 sae mutant was obtained by screening a Tn917 library of the agr mutant ISP479Mu for clones with altered hemolytic activity. Sequence analysis of the sae operon revealed two additional open reading frames (ORFs) (ORF3 and ORF4) upstream of the two-component regulatory genes saeR and saeS. Four overlapping sae-specific transcripts (T1 to T4) were detected by Northern blot analysis, and the transcriptional initiation points were mapped by primer extension analysis. The T1, T2, and T3 mRNAs are probably terminated at the same stem-loop sequence downstream of saeS. The T1 message (3.1 kb) initiates upstream of ORF4, T2 (2.4 kb) initiates upstream of ORF3, and T3 (2.0 kb) initiates in front of saeR. T4 (0.7 kb) represents a monocistronic mRNA encompassing ORF4 only. sae-specific transcripts were detectable in all of the 40 different clinical S. aureus isolates investigated. Transcript levels were at maximum during the post-exponential growth phase. The sae mutant showed a significantly reduced rate of invasion of human endothelial cells, consistent with diminished transcription and expression of fnbA. The expression of type 5 capsular polysaccharide is activated in the sae mutant of strain Newman, as shown by immunofluorescence and promoter-reporter fusion experiments. In summary, the sae operon constitutes a four-component regulator system which acts on virulence gene expression in S. aureus. PMID- 14563863 TI - Molecular characterization of the Mg2+-responsive PhoP-PhoQ regulon in Salmonella enterica. AB - The PhoP/PhoQ two-component system controls the extracellular magnesium deprivation response in Salmonella enterica. In addition, several virulence associated genes that are mainly required for intramacrophage survival during the infection process are under the control of its transcriptional regulation. Despite shared Mg(2+) modulation of the expression of the PhoP-activated genes, no consensus sequence common to all of them could be detected in their promoter regions. We have investigated the transcriptional regulation and the interaction of the response regulator PhoP with the promoter regions of the PhoP-activated loci phoPQ, mgtA, slyB, pmrD, pcgL, phoN, pagC, and mgtCB. A direct repeat of the heptanucleotide sequence (G/T)GTTTA(A/T) was identified as the conserved motif recognized by PhoP to directly control the gene expression of the first five loci, among which the first four are ancestral to enterobacteria. On the other hand, no direct interaction of the response regulator with the promoter of phoN, pagC, or mgtCB was apparent by either in vitro or in vivo assays. These loci are Salmonella specific and were probably acquired by horizontal DNA transfer. Besides, sequence analysis of pag promoters revealed the presence of a conserved PhoP box in 6 out of the 12 genes analyzed. Our results strongly suggest that the expression of a set of Mg(2+)-controlled genes is driven by PhoP via unknown intermediate regulatory mechanisms that could also involve ancillary factors. PMID- 14563864 TI - Requirement of the Listeria monocytogenes broad-range phospholipase PC-PLC during infection of human epithelial cells. AB - In this study, we investigated the requirement of the Listeria monocytogenes broad-range phospholipase C (PC-PLC) during infection of human epithelial cells. L. monocytogenes is a facultative intracellular bacterial pathogen of humans and a variety of animal species. After entering a host cell, L. monocytogenes is initially surrounded by a membrane-bound vacuole. Bacteria promote their escape from this vacuole, grow within the host cell cytosol, and spread from cell to cell via actin-based motility. Most infection studies with L. monocytogenes have been performed with mouse cells or an in vivo mouse model of infection. In all mouse-derived cells tested, the pore-forming cytolysin listeriolysin O (LLO) is absolutely required for lysis of primary vacuoles formed during host cell entry. However, L. monocytogenes can escape from primary vacuoles in the absence of LLO during infection of human epithelial cell lines Henle 407, HEp-2, and HeLa. Previous studies have shown that the broad-range phospholipase C, PC-PLC, promotes lysis of Henle 407 cell primary vacuoles in the absence of LLO. Here, we have shown that PC-PLC is also required for lysis of HEp-2 and HeLa cell primary vacuoles in the absence of LLO expression. Furthermore, our results indicated that the amount of PC-PLC activity is critical for the efficiency of vacuolar lysis. In an LLO-negative derivative of L. monocytogenes strain 10403S, expression of PC-PLC has to increase before or upon entry into human epithelial cells, compared to expression in broth culture, to allow bacterial escape from primary vacuoles. Using a system for inducible PC-PLC expression in L. monocytogenes, we provide evidence that phospholipase activity can be increased by elevated expression of PC-PLC or Mpl, the enzyme required for proteolytic activation of PC-PLC. Lastly, by using the inducible PC-PLC expression system, we demonstrate that, in the absence of LLO, PC-PLC activity is not only required for lysis of primary vacuoles in human epithelial cells but is also necessary for efficient cell-to-cell spread. We speculate that the additional requirement for PC-PLC activity is for lysis of secondary double-membrane vacuoles formed during cell-to-cell spread. PMID- 14563865 TI - A mutant of Paracoccus denitrificans with disrupted genes coding for cytochrome c550 and pseudoazurin establishes these two proteins as the in vivo electron donors to cytochrome cd1 nitrite reductase. AB - In Paracoccus denitrificans, electrons pass from the membrane-bound cytochrome bc(1) complex to the periplasmic nitrite reductase, cytochrome cd(1). The periplasmic protein cytochrome c(550) has often been implicated in this electron transfer, but its absence, as a consequence of mutation, has previously been shown to result in almost no attenuation in the ability of the nitrite reductase to function in intact cells. Here, the hypothesis that cytochrome c(550) and pseudoazurin are alternative electron carriers from the cytochrome bc(1) complex to the nitrite reductase was tested by construction of mutants of P. denitrificans that are deficient in either pseudoazurin or both pseudoazurin and cytochrome c(550). The latter organism, but not the former (which is almost indistinguishable in this respect from the wild type), grows poorly under anaerobic conditions with nitrate as an added electron acceptor and accumulates nitrite in the medium. Growth under aerobic conditions with either succinate or methanol as the carbon source is not significantly affected in mutants lacking either pseudoazurin or cytochrome c(550) or both these proteins. We concluded that pseudoazurin and cytochrome c(550) are the alternative electron mediator proteins between the cytochrome bc(1) complex and the cytochrome cd(1)-type nitrite reductase. We also concluded that expression of pseudoazurin is mainly controlled by the transcriptional activator FnrP. PMID- 14563866 TI - Increasing the ratio of Soj to Spo0J promotes replication initiation in Bacillus subtilis. AB - The ParA and ParB protein families are well conserved in bacteria. However, their functions are still unclear. In Bacillus subtilis, Soj and Spo0J are members of these two protein families, respectively. A previous report revealed that replication initiated early and asynchronously in spo0J null mutant cells, as determined by flow cytometry. In this study, we examined the cause of this promotion of replication initiation. Deletion of both the soj and spo0J genes restored the frequency of replication initiation to almost the wild-type level, suggesting that production of Soj in the absence of Spo0J leads to early and asynchronous initiation of replication. Consistent with this suggestion, overproduction of Soj in wild-type cells had the same effect on replication initiation as in the spo0J null mutant, and overproduction of both Soj and Spo0J did not. These results indicate that when the ratio of Soj to Spo0J increases, Soj interferes with tight control of replication initiation and causes early and asynchronous initiation. Whereas replication initiation also occurred significantly earlier in the two spo0J mutants, spo0J14 and spo0J17, it occurred only slightly early in the sojK16Q mutant and was delayed in the sojG12V mutant. Although Soj localized to nucleoids in the spo0J mutants, the two Soj mutant proteins were distributed throughout the cell or localized to cell poles. Thus, interestingly, the promotion of replication initiation seems to correlate with localization of Soj to nucleoids. This may suggest that Soj inhibits transcription of some cell cycle genes and leads to early and asynchronous initiation of replication. In wild-type cells Spo0J counteracts this Soj function. PMID- 14563867 TI - Characterization of the integrase gene and attachment site for the Myxococcus xanthus bacteriophage Mx9. AB - Bacteriophage Mx9 is a temperate phage that infects Myxococcus xanthus. It lysogenizes the bacteria by integrating into the bacterial chromosome by site specific recombination at one of two sites, attB1 or attB2. Integration at attB1 results in deletion of DNA between the two attB sites. The attB2 site lies within the 5' region of the M. xanthus tRNA(Gly) gene. Mx9 integration requires a single protein, Int. Analysis of integration revealed that the phage attachment site (attP) is contained in the int gene and that upon integration, the 3' end of the int gene is altered. Plasmids containing fusions of the pilA or mgl promoter to lacZ integrated at either Mx9 attB site have higher levels of transcription than the same fusions integrated at the Mx8 attB site. PMID- 14563868 TI - A new heat shock gene, AgsA, which encodes a small chaperone involved in suppressing protein aggregation in Salmonella enterica serovar typhimurium. AB - We discovered a novel small heat shock protein (sHsp) named AgsA (aggregation suppressing protein) in the thermally aggregated fraction from a Salmonella enterica serovar Typhimurium dnaK-null strain. The -10 and -35 regions upstream of the transcriptional start site of the agsA gene are characteristic of sigma(32)- and sigma(72)-dependent promoters. AgsA was strongly induced by high temperatures. The similarity between AgsA and the other two sHsps of Salmonella serovar Typhimurium, IbpA and IbpB, is rather low (around 30% amino acid sequence identity). Phylogenetic analysis suggested that AgsA arose from an ancient gene duplication or amplification at an early evolutionary stage of gram-negative bacteria. Here we show that overproduction of AgsA partially complements the DeltadnaK52 thermosensitive phenotype and reduces the amount of heat-aggregated proteins in both DeltadnaK52 and DeltarpoH mutants of Escherichia coli. These data suggest that AgsA is an effective chaperone capable of preventing aggregation of nonnative proteins and maintaining them in a state competent for refolding in Salmonella serovar Typhimurium at high temperatures. PMID- 14563869 TI - Biochemical properties and regulated gene expression of the superoxide dismutase from the facultatively aerobic hyperthermophile Pyrobaculum calidifontis. AB - Superoxide dismutase (SOD) was purified from a facultatively aerobic hyperthermophilic archaeon, Pyrobaculum calidifontis VA1. The purified native protein from aerobically grown cells exhibited 1,960 U of SOD activity/mg and contained 0.86 +/- 0.04 manganese and <0.01 iron atoms per subunit. The gene encoding SOD was cloned and expressed in Escherichia coli. Although the recombinant protein was soluble, little activity was observed due to the lack of metal incorporation. Reconstitution of the enzyme by heat treatment with either Mn or Fe yielded a highly active protein with specific activities of 1,970 and 434 U/mg, respectively. This indicated that the SOD from P. calidifontis was a cambialistic SOD with a preference toward Mn in terms of activity. Interestingly, reconstitution experiments in vitro indicated a higher tendency of the enzyme to incorporate Fe than Mn. When P. calidifontis was grown under anaerobic conditions, a majority of the native SOD was incorporated with Fe, indicating the cambialistic property of this enzyme in vivo. We further examined the expression levels of SOD and a previously characterized Mn catalase from this strain in the presence or absence of oxygen. Northern blot, Western blot, and activity measurement analyses revealed that both genes are expressed at much higher levels under aerobic conditions. We also detected a rapid response in the biosynthesis of these enzymes once the cells were exposed to oxygen. PMID- 14563870 TI - Recognition of DNA by three ferric uptake regulator (Fur) homologs in Bacillus subtilis. AB - Bacillus subtilis contains three Fur homologs: Fur, PerR, and Zur. Despite significant sequence similarities, they respond to different stimuli and regulate different sets of genes. DNA target site comparisons indicate that all three paralogs recognize operators with a core 7-1-7 inverted repeat. The corresponding consensus sequences are identical at five or more of the seven defined positions. Using site-directed mutagenesis, the Per box at the mrgA promoter was altered to mimic the core 7-1-7 motif of the Fur and Zur boxes. In vitro, the mrgA promoter containing a Zur box was only recognized by Zur, as demonstrated by DNase I footprinting assays. In contrast, both Fur and PerR bound to the mrgA promoter region containing a consensus Fur box. Expression analysis of these promoters is consistent with the in vitro data demonstrating as few as 1 or 2 base changes per half-site are sufficient to alter regulation. Similarly, the Fur box at the feuA promoter can be converted into a Per or a Zur box by appropriate mutations. While both Fur and PerR could recognize some of the same synthetic operator sequences, no naturally occurring sites are known that are subject to dual regulation. However, the PerR-regulated zosA gene is controlled from a regulatory region that contains both Per and Fur boxes. Although purified Fur protein bound to the candidate Fur boxes, Fur has little effect on zosA expression-possibly due to the location of the Fur boxes relative to the zosA promoter. Together, our results identify two nucleotide positions that are important for the ability of PerR, Fur, and Zur to distinguish among the many closely related operator sites present in the B. subtilis genome. PMID- 14563872 TI - The IS1111 family members IS4321 and IS5075 have subterminal inverted repeats and target the terminal inverted repeats of Tn21 family transposons. AB - IS5075 and IS4321 are closely related (93.1% identical) members of the IS1111 family that target a specific position in the 38-bp terminal inverted repeats of Tn21 family transposons and that are inserted in only one orientation. They are 1,327 bp long and have identical ends consisting of short inverted repeats of 12 bp with an additional 7 bp (TAATGAG) or 6 bp (AATGAG) to the left of the left inverted repeats and 3 bp (AGA) or 4 bp (AGAT) to the right of the right inverted repeat. Circular forms of IS5075 and IS4321 in which the inverted repeats are separated by abutting terminal sequences (AGATAATGAG) were detected. A similar circular product was found for the related ISPa11. Transposition of IS4321 into the 38-bp target site was detected, but a flanking duplication was not generated. The precisely reconstituted target site was also identified. Over 50 members of the IS1111 family were identified. They encode related transposases, have related inverted repeats, and include related bases that lie outside these inverted repeats. In some, the flanking bases number 5 or 6 on the left and 4 or 3 on the right. Specific target sites were found for several of these insertion sequence (IS) elements. IS1111 family members therefore differ from the majority of IS elements, which are characterized by terminal inverted repeats and a target site duplication, and from members of the related IS110 family, which do not have obvious inverted repeats near their termini. PMID- 14563871 TI - Genome-wide transcriptional profiling analysis of adaptation of Bacillus subtilis to high salinity. AB - The gram-positive soil bacterium Bacillus subtilis often faces increases in the salinity in its natural habitats. A transcriptional profiling approach was utilized to investigate both the initial reaction to a sudden increase in salinity elicited by the addition of 0.4 M NaCl and the cellular adaptation reactions to prolonged growth at high salinity (1.2 M NaCl). Following salt shock, a sigB mutant displayed immediate and transient induction and repression of 75 and 51 genes, respectively. Continuous propagation of this strain in the presence of 1.2 M NaCl triggered the induction of 123 genes and led to the repression of 101 genes. In summary, our studies revealed (i) an immediate and transient induction of the SigW regulon following salt shock, (ii) a role of the DegS/DegU two-component system in sensing high salinity, (iii) a high-salinity mediated iron limitation, and (iv) a repression of chemotaxis and motility genes by high salinity, causing severe impairment of the swarming capability of B. subtilis cells. Initial adaptation to salt shock and continuous growth at high salinity share only a limited set of induced and repressed genes. This finding strongly suggests that these two phases of adaptation require distinctively different physiological adaptation reactions by the B. subtilis cell. The large portion of genes with unassigned functions among the high-salinity-induced or repressed genes demonstrates that major aspects of the cellular adaptation of B. subtilis to high salinity are unexplored so far. PMID- 14563873 TI - Investigation of the role of electrostatic charge in activation of the Escherichia coli response regulator CheY. AB - In a two-component regulatory system, an important means of signal transduction in microorganisms, a sensor kinase phosphorylates a response regulator protein on an aspartyl residue, resulting in activation. The active site of the response regulator is highly charged (containing a lysine, the phosphorylatable aspartate, two additional aspartates involved in metal binding, and an Mg(2+) ion), and introduction of the dianionic phosphoryl group results in the repositioning of charged moieties. Furthermore, substitution of one of the Mg(2+)-coordinating aspartates with lysine or arginine in the Escherichia coli chemotaxis response regulator CheY results in phosphorylation-independent activation. In order to examine the consequences of altered charge distribution for response regulator activity and to identify possible additional amino acid substitutions that result in phosphorylation-independent activation, we made 61 CheY mutants in which residues close to the site of phosphorylation (Asp57) were replaced by various charged amino acids. Most substitutions (47 of 61) resulted in the complete loss of CheY activity, as measured by the inability to support clockwise flagellar rotation. However, 10 substitutions, all introducing a new positive charge, resulted in the loss of chemotaxis but in the retention of some clockwise flagellar rotation. Of the mutants in this set, only the previously identified CheY13DK and CheY13DR mutants displayed clockwise activity in the absence of the CheA sensor kinase. The absence of negatively charged substitution mutants with residual activity suggests that the introduction of additional negative charges into the active site is particularly deleterious for CheY function. Finally, the spatial distribution of positions at which amino acid substitutions are functionally tolerated or not tolerated is consistent with the presently accepted mechanism of response regulator activation and further suggests a possible role for Met17 in signal transduction by CheY. PMID- 14563874 TI - Genome-scale analysis of the uses of the Escherichia coli genome: model-driven analysis of heterogeneous data sets. AB - The recent availability of heterogeneous high-throughput data types has increased the need for scalable in silico methods with which to integrate data related to the processes of regulation, protein synthesis, and metabolism. A sequence-based framework for modeling transcription and translation in prokaryotes has been established and has been extended to study the expression state of the entire Escherichia coli genome. The resulting in silico analysis of the expression state highlighted three facets of gene expression in E. coli: (i) the metabolic resources required for genome expression and protein synthesis were found to be relatively invariant under the conditions tested; (ii) effective promoter strengths were estimated at the genome scale by using global mRNA abundance and half-life data, revealing genes subject to regulation under the experimental conditions tested; and (iii) large-scale genome location-dependent expression patterns with approximately 600-kb periodicity were detected in the E. coli genome based on the 49 expression data sets analyzed. These results support the notion that a structured model-driven analysis of expression data yields additional information that can be subjected to commonly used statistical analyses. The integration of heterogeneous genome-scale data (i.e., sequence, expression data, and mRNA half-life data) is readily achieved in the context of an in silico model. PMID- 14563875 TI - Description and interpretation of adaptive evolution of Escherichia coli K-12 MG1655 by using a genome-scale in silico metabolic model. AB - Genome-scale in silico metabolic networks of Escherichia coli have been reconstructed. By using a constraint-based in silico model of a reconstructed network, the range of phenotypes exhibited by E. coli under different growth conditions can be computed, and optimal growth phenotypes can be predicted. We hypothesized that the end point of adaptive evolution of E. coli could be accurately described a priori by our in silico model since adaptive evolution should lead to an optimal phenotype. Adaptive evolution of E. coli during prolonged exponential growth was performed with M9 minimal medium supplemented with 2 g of alpha-ketoglutarate per liter, 2 g of lactate per liter, or 2 g of pyruvate per liter at both 30 and 37 degrees C, which produced seven distinct strains. The growth rates, substrate uptake rates, oxygen uptake rates, by product secretion patterns, and growth rates on alternative substrates were measured for each strain as a function of evolutionary time. Three major conclusions were drawn from the experimental results. First, adaptive evolution leads to a phenotype characterized by maximized growth rates that may not correspond to the highest biomass yield. Second, metabolic phenotypes resulting from adaptive evolution can be described and predicted computationally. Third, adaptive evolution on a single substrate leads to changes in growth characteristics on other substrates that could signify parallel or opposing growth objectives. Together, the results show that genome-scale in silico metabolic models can describe the end point of adaptive evolution a priori and can be used to gain insight into the adaptive evolutionary process for E. coli. PMID- 14563876 TI - Analysis of specific binding involved in genomic packaging of the double-stranded RNA bacteriophage phi6. AB - The genomes of bacteriophage phi6 and its relatives are packaged through a mechanism that involves the recognition and translocation of the three different plus-strand transcripts of the segmented double-stranded-RNA genomes into preformed polyhedral structures called procapsids or inner cores. The packaging requires the hydrolysis of nucleoside triphosphates and takes place in the order segment S-segment M, segment L. Packaging is dependent upon unique sequences of about 200 nucleotides near the 5' ends of plus-strand transcripts of the three genomic segments. It appears that P1 is the determinant of the RNA binding sites. Directed mutation of P1 was used to locate regions that are important for genomic packaging. Specific binding of RNA to the exterior of the procapsid was dependent upon ATP, and a region that showed a high level of cross-linking to phage specific RNA was located. Antibodies to peptide sequences were prepared, and their abilities to bind to the exterior of procapsids were determined. Sites sensitive to trypsin and to factor Xa were determined as well. PMID- 14563877 TI - Why is carbonic anhydrase essential to Escherichia coli? AB - The can (previously yadF) gene of Escherichia coli encodes a beta-class carbonic anhydrase (CA), an enzyme which interconverts CO(2) and bicarbonate. Various essential metabolic processes require either CO(2) or bicarbonate and, although carbon dioxide and bicarbonate spontaneously equilibrate in solution, the low concentration of CO(2) in air and its rapid diffusion from the cell mean that insufficient bicarbonate is spontaneously made in vivo to meet metabolic and biosynthetic needs. We calculate that demand for bicarbonate is 10(3)- to 10(4) fold greater than would be provided by uncatalyzed intracellular hydration and that enzymatic conversion of CO(2) to bicarbonate is therefore necessary for growth. We find that can expression is ordinarily required for growth in air. It is dispensable if the atmospheric partial pressure of CO(2) is high or during anaerobic growth in a closed vessel at low pH, where copious CO(2) is generated endogenously. CynT, the single E. coli Can paralog, can, when induced with azide, replace Can; also, the gamma-CA from Methanosarcina thermophila can at least partially replace it. Expression studies showed that can transcription does not appear to respond to carbon dioxide concentration or to be autoregulated. However, can expression is influenced by growth rate and the growth cycle; it is expressed best in slow-growing cultures and at higher culture densities. Expression can vary over a 10-fold range during the growth cycle and is also elevated during starvation or heat stress. PMID- 14563878 TI - Isolation and characterization of mutants of the Bacillus subtilis oligopeptide permease with altered specificity of oligopeptide transport. AB - Bacterial oligopeptide permeases are members of the large family of ATP binding cassette transporters and typically import peptides of 3 to 5 amino acids, apparently independently of sequence. Oligopeptide permeases are needed for bacteria to utilize peptides as nutrient sources and are sometimes involved in signal transduction pathways. The Bacillus subtilis oligopeptide permease stimulates competence development and the initiation of sporulation, at least in part, by importing specific signaling peptides. We isolated rare, partly functional mutations in B. subtilis opp. The mutants were resistant to a toxic tripeptide but still retained the ability to sporulate and/or become competent. The mutations, mostly in the oligopeptide binding protein located on the cell surface, affected residues whose alteration appears to change the specificity of oligopeptide transport. PMID- 14563879 TI - Genome sequences of two closely related Vibrio parahaemolyticus phages, VP16T and VP16C. AB - Two bacteriophages of an environmental isolate of Vibrio parahaemolyticus were isolated and sequenced. The VP16T and VP16C phages were separated from a mixed lysate based on plaque morphology and exhibit 73 to 88% sequence identity over about 80% of their genomes. Only about 25% of their predicted open reading frames are similar to genes with known functions in the GenBank database. Both phages have cos sites and open reading frames encoding proteins closely related to coliphage lambda's terminase protein (the large subunit). Like in coliphage lambda and other siphophages, a large operon in each phage appears to encode proteins involved in DNA packaging and capsid assembly and presumably in host lysis; we refer to this as the structural operon. In addition, both phages have open reading frames closely related to genes encoding DNA polymerase and helicase proteins. Both phages also encode several putative transcription regulators, an apparent polypeptide deformylase, and a protein related to a virulence-associated protein, VapE, of Dichelobacter nodosus. Despite the similarity of the proteins and genome organization, each of the phages also encodes a few proteins not encoded by the other. We did not identify genes closely related to genes encoding integrase proteins belonging to either the tyrosine or serine recombinase family, and we have no evidence so far that these phages can lysogenize the V. parahaemolyticus strain 16 host. Surprisingly for active lytic viruses, the two phages have a codon usage that is very different than that of the host, suggesting the possibility that they may be relative newcomers to growth in V. parahaemolyticus. The DNA sequences should allow us to characterize the lifestyles of VP16T and VP16C and the interactions between these phages and their host at the molecular level, as well as their relationships to other marine and nonmarine phages. PMID- 14563880 TI - The gene yjcG, cotranscribed with the gene acs, encodes an acetate permease in Escherichia coli. AB - We isolated an Escherichia coli mutant strain that suppresses the glycolate negative phenotype of a strain deficient in both GlcA and LldP transporters of this compound. This suppressing phenotype was assigned to yjcG, a gene whose function was previously unknown, which was found to encode a membrane protein able to transport glycolate. On the basis of sequence similarity, the yjcG gene product was classified as a member of the sodium:solute symporter family. Northern experiments revealed that yjcG is cotranscribed with its neighbor, acs, encoding acetyl coenzyme A synthetase, which is involved in the scavenging acetate. The fortuitous presence of an IS2 element in acs, which impaired yjcG expression by polarity in our parental strain, allowed us to conclude that the alternative glycolate carrier became active after precise excision of IS2 in the suppressed strain. The finding that yjcG encodes a putative membrane carrier for glycolate and the cotranscription of yjcG with acs suggested that the primary function of the yjcG gene product (proposed gene name, actP) could be acetate transport and allowed us to define an operon involved in acetate metabolism. The time course of [1,2-(14)C]acetate uptake and the results of a concentration kinetics analysis performed with cells expressing ActP or cells deficient in ActP supported the the hypothesis that this carrier is an acetate transporter and suggested that there may be another transport system for this monocarboxylate. PMID- 14563881 TI - Identification of quorum-sensing-regulated genes of Burkholderia cepacia. AB - Quorum sensing is a regulatory mechanism (operating in response to cell density) which in gram-negative bacteria usually involves the production of N-acyl homoserine lactones (HSL). Quorum sensing in Burkholderia cepacia has been associated with the regulation of expression of extracellular proteins and siderophores and also with the regulation of swarming and biofilm formation. In the present study, several quorum-sensing-controlled gene promoters of B. cepacia ATCC 25416 were identified and characterized. A total of 28 putative gene promoters show CepR-C(8)-HSL-dependent expression, suggesting that quorum sensing in B. cepacia is a global regulatory system. PMID- 14563882 TI - Nucleotide sequence of the integration site of the temperate bacteriophage 6220, which carries the Shiga toxin gene stx(1ox3). AB - The integration site, attR, of the Shiga toxin-encoding phage 6220 (stx(1ox3)) has been determined. The phage integrates into the chromosome of its Escherichia coli host strain, CB6220, within a gene that is homologous to gene Z2577 and encodes an oxidoreductase. This new integration site was found in different Stx1ox3-producing enterohemorrhagic E. coli strains, which were analyzed by PCR. PMID- 14563883 TI - Prophage contribution to bacterial population dynamics. AB - Cocultures of Salmonella strains carrying or lacking specific prophages undergo swift composition changes as a result of phage-mediated killing of sensitive bacteria and lysogenic conversion of survivors. Thus, spontaneous prophage induction in a few lysogenic cells enhances the competitive fitness of the lysogen population as a whole, setting a selection regime that forces maintenance and spread of viral DNA. This is likely to account for the profusion of prophage sequences in bacterial genomes and may contribute to the evolutionary success of certain phylogenetic lineages. PMID- 14563884 TI - A transcriptional pause synchronizes translation with transcription in the tryptophanase operon leader region. AB - Regulation of transcription of the tryptophanase operon requires that translation of its leader peptide coding region, tnaC, be coupled with its transcription. We show in vitro that a transcription pause site exists at the end of the tnaC coding region and that translation of tnaC releases the paused transcription complex, coupling transcription with translation. PMID- 14563886 TI - Bacteriophage ST64B, a genetic mosaic of genes from diverse sources isolated from Salmonella enterica serovar typhimurium DT 64. AB - The complete sequence of the double-stranded DNA (dsDNA) genome of the Salmonella enterica serovar Typhimurium ST64B bacteriophage was determined. The 40,149-bp genomic sequence of ST64B has an overall G+C content of 51.3% and is distinct from that of P22. The genome architecture is similar to that of the lambdoid phages, particularly that of coliphage lambda. Most of the putative tail genes showed sequence similarity to tail genes of Mu, a nonlambdoid phage. In addition, it is likely that these tail genes are not expressed due to insertions of fragments of genes related to virulence within some of the open reading frames. This, together with the inability of ST64B to produce plaques on a wide range of isolates, suggests that ST64B is a defective phage. In contrast to the tail genes, most of the head genes showed similarity to those of the lambdoid phages HK97 and HK022, but these head genes also have significant sequence similarities to those of several other dsDNA phages infecting diverse bacterial hosts, including Escherichia, Pseudomonas, Agrobacterium, Caulobacter, Mesorhizobium, and Streptomyces: This suggests that ST64B is a genetic mosaic that has acquired significant portions of its genome from sources outside the genus Salmonella. PMID- 14563885 TI - Thymine at -5 is crucial for cpc promoter activity of Synechocystis sp. strain PCC 6714. AB - The levels of transcripts of the cpc operon were highly reduced in a PD-1 mutant of cyanobacterium Synechocystis sp. strain PCC 6714. This was due to a substitution of C for T that occurred at 5 bp upstream of the transcription initiation site of the cpc operon. Any substitution for T at the -5 position drastically reduced both in vivo and in vitro promoter activity in cyanobacterium Synechococcus sp. strain PCC 7942 but not the in vivo activity in Escherichia coli. This suggests that the requirement of -5T appears to be specific for a cyanobacterial RNA polymerase-promoter combination. PMID- 14563887 TI - rRNA antitermination functions with heat shock promoters. AB - Transcription antitermination in the rRNA operons of Escherichia coli requires a unique nucleic acid sequence that serves as a signal for modification of the elongating RNA polymerase, making it resistant to Rho-dependent termination. We examined the antitermination ability of RNA polymerase elongation complexes that had initiated at three different heat shock promoters, dnaK, groE, and clpB, and then transcribed the antitermination sequence to read through a Rho-dependent terminator. Terminator bypass comparable to that seen with sigma(70) promoters was obtained. Lack of or inversion of the sequence abolished terminator readthrough. We conclude that RNA polymerase that uses sigma(32) to initiate transcription can adopt a conformation similar to that of sigma(70)-containing RNA polymerase, enabling it to interact with auxiliary modifying proteins and bypass Rho-dependent terminators. PMID- 14563888 TI - Host range of chlamydiaphages phiCPAR39 and Chp3. AB - The host range of phiCPAR39 is limited to four Chlamydophila species: C. abortus, C. caviae, C. pecorum, and C. pneumoniae. Chp3 (a newly discovered bacteriophage isolated from C. pecorum) shares three of these hosts (C. abortus, C. caviae, and C. pecorum) but can additionally infect Chlamydophila felis. The ability to support replication was directly correlated with the binding properties of the respective bacteriophages with their host species. Binding studies also show that phiCPAR39 and Chp3 use different host receptors to infect the same host cells: cell binding is sensitive to proteinase K treatment, confirming that the chlamydiaphage receptors are proteinaceous in nature. PMID- 14563889 TI - Effect of extended perioperative antibiotic prophylaxis on intravascular catheter colonization and infection in cardiothoracic surgery patients. AB - Intravascular catheter-related infections (CRI) account for one third of nosocomial bloodstream infections in England. UK Department of Health guidelines state that antibiotic prophylaxis is not required during placement, or use of, central venous catheters, to prevent CRI. However, some clinicians continue to use antibiotics in an attempt to prevent CRI. We investigated the effect of extended routine perioperative antibiotic prophylaxis in cardiothoracic patients on rates of intravascular catheter (IVC) colonization and infection. Investigations were undertaken in patients undergoing uncomplicated cardiothoracic surgery during July 2001-February 2002. Patients who received three doses of cefuroxime as perioperative prophylaxis were compared with those who received extended cefuroxime prophylaxis until the IVC was removed. Patients were not randomized into groups, but received the different prophylaxis regimens according to the usual practice of the consultant cardiothoracic surgeon. A roll tip method was used to determine IVC colonization. Of 191 patients who fulfilled the inclusion criteria, 12 were excluded because data were incomplete. One hundred and forty-six patients received routine prophylaxis, and 33 prophylaxis until the IVC was removed. Twenty-three out of 146 (16%) IVCs in the 'routine' group and four out of 33 (12%) in the 'extended' group became colonized; no IVC related bloodstream infections occurred during the survey. The duration of IVC placement and the types of operation performed in the two groups were not significantly different (P > 0.05). In routine cardiothoracic surgery patients, extending routine perioperative antibiotic prophylaxis until all IVCs have been removed does not influence rates of IVC colonization. PMID- 14563890 TI - Antimycobacterial activity of 2-methyl-adenosine. AB - OBJECTIVES: The aims of this study were to assess the in vitro activity of 2 methyl-adenosine against Mycobacterium tuberculosis and evaluate, and to intracellular efficacy, and to evaluate its effectiveness against M. tuberculosis in a persistent state model and examine its potential mechanism of action. METHODS: In vitro activity was determined by means of a colorimetric microdilution broth assay. Intracellular activity was assessed with a Mono Mac 6 human monocytic cell line. A hypoxic shift-down model was used to evaluate the effect of 2-methyl-adenosine on M. tuberculosis in a persistent state. Mechanism of-action studies were conducted by examining the effect of 2-methyl-adenosine on the uptake of appropriate radiolabelled precursors into respective mycobacterial macromolecular components. RESULTS: Studies confirmed the in vitro activity of 2 methyl-adenosine against M. tuberculosis and demonstrated intracellular efficacy against M. tuberculosis within macrophages. 2-Methyl-adenosine was able to significantly affect the viability of M. tuberculosis in a hypoxic shift-down model previously described to simulate the persistent state that results during tuberculosis. Mechanism-of-action studies revealed that the immediate inhibitory effects of 2-methyl-adenosine were associated with protein and DNA synthesis and not RNA synthesis. CONCLUSIONS: Results indicate that 2-methyl-adenosine, or similar derivatives, might be effective against M. tuberculosis infections during latency. This information should be helpful in understanding purine metabolism of M. tuberculosis and also the metabolic activity of this important human pathogen in the persistent state. PMID- 14563891 TI - Mode of action of pyrazinamide: disruption of Mycobacterium tuberculosis membrane transport and energetics by pyrazinoic acid. AB - Pyrazinamide is an important sterilizing drug that shortens tuberculosis (TB) therapy. However, the mechanism of action of pyrazinamide is poorly understood because of its unusual properties. Here we show that pyrazinoic acid, the active moiety of pyrazinamide, disrupted membrane energetics and inhibited membrane transport function in Mycobacterium tuberculosis. The preferential activity of pyrazinamide against old non-replicating bacilli correlated with their low membrane potential and the disruption of membrane potential by pyrazinoic acid and acid pH. Inhibitors of membrane energetics increased the antituberculous activity of pyrazinamide. These findings shed new light on the mode of action of pyrazinamide and may help in the design of new drugs that shorten therapy. PMID- 14563892 TI - Inhibition of Streptococcus mutans biofilm accumulation and polysaccharide production by apigenin and tt-farnesol. AB - OBJECTIVES: Apigenin is a potent inhibitor of glucosyltransferases and tt farnesol affects the membrane integrity of Streptococcus mutans. We investigated the influence of apigenin and tt-farnesol, alone and in combination, on the accumulation, polysaccharide composition and viability of S. mutans UA159 biofilms. METHODS: Initially, biofilms were grown for 54 h; then, the early formed biofilms were treated for 1 min twice daily with one of the following: (i). 1.33 mM tt-farnesol; (ii). 1.33 mM apigenin; (iii). apigenin + tt-farnesol (1.33 mM each); (iv). vehicle control (20% ethanol with 0.75% dimethyl sulphoxide); (v). 0.12% chlorhexidine (1.33 mM); or (vi). physiological saline (145 mM NaCl). The procedure was repeated at biofilm ages of 78 and 102 h, and biofilms were harvested at 126 h. The dry weight, protein concentration, number of cfu, and polysaccharide composition per biofilm were determined. RESULTS: The dry weights of the biofilms treated with the test agents were significantly less (30-50%) than those treated with vehicle control (P < 0.05). Biofilms treated with the test agents also resulted in lower amounts of extracellular alkali soluble glucans, intracellular iodophilic polysaccharides and, to a lesser extent, fructans. The fructosyltransferase activity was affected only by apigenin and apigenin + tt-farnesol. The recoverable viable counts of S. mutans were slightly lower (0.5 to 1 log10 decrease in cfu/biofilm) after apigenin and tt farnesol treatments compared with the vehicle control. Chlorhexidine displayed potent bactericidal activity, and virtually halted the further accumulation of early-formed (54 h old) biofilms. CONCLUSIONS: Apigenin and tt-farnesol affected the accumulation and polysaccharide content of S. mutans biofilms without major impact on the bacterial viability. PMID- 14563893 TI - Prevention of invasive fungal infections in liver transplant recipients: the role of prophylaxis with lipid formulations of amphotericin B in high-risk patients. AB - BACKGROUND: Invasive fungal infections (IFI) are associated with high mortality in liver transplant recipients. Prevention remains an elusive goal, especially for IFI caused by moulds. PATIENTS AND METHODS: From January 1998, patients who fulfilled four or more variables identified as risk factors for IFI received a cumulative dose of 1-1.5 g of lipid formulations of amphotericin B (L-AmpB; AmBisome or Abelcet). The development of IFI in these patients was compared with historical patients. RESULTS: Two hundred and eighty liver transplant recipients were analysed over a period of 8 years. In the historical group, IFI were observed in 22 of 131 patients (17%) and invasive aspergillosis in 13 of them (10%). After January 1998, IFI were observed in nine of 149 (6%) (P < 0.01) and invasive aspergillosis in six patients (4%) (P = 0.08). In patients with four or more risk factors (high risk) for IFI, the administration of L-AmpB reduced the risk from 36% to 14% (P = 0.07), and the risk of aspergillosis from 23% to 5% (P = 0.08). Notably, prophylaxis reduced the risk of aspergillosis from 32% to 0% in dialysed patients (P = 0.03). Variables independently associated with IFI in high risk patients were dialysis [odds ratio (OR) 3.9; 95% confidence interval (CI) 1 16.7] and surgical reintervention (OR 5.4; 95% CI 1.2-24.6), while L-AmpB was a protective factor in this multivariate analysis (OR 0.1; 95% CI 0.02-0.8). The analysis in these high-risk patients was not able to demonstrate an association between the administration of L-AmpB and higher survival. CONCLUSIONS: Selected risk factors are good predictors of IFI in liver transplant recipients. The administration of L-AmpB in high-risk patients is independently associated with a reduction of IFI. PMID- 14563894 TI - Chloramphenicol-resistant Neisseria meningitidis containing catP isolated in Australia. AB - Earlier workers have described chloramphenicol resistance in meningococci isolated from cerebrospinal fluid sampled in patients in Vietnam (11 cases) and France (one case) during 1987-1996. Here we describe two distinct serogroup B strains isolated in Australia in 1994 and 1997, and found among approximately 1400 invasive meningococcal isolates examined in Australia over a 9 year period. Both were phenotypically chloramphenicol resistant on disc, Etest and agar inclusion MIC and acetylated chloramphenicol examination. DNA amplification and sequencing confirmed the presence of catP and the 3' end of tnpV from Tn4451, a mobilizable element from Clostridium perfringens, although other sequences were not present. Tn4451 has inserted into a gene designated TIGR locus NMB1350 in both isolates with no loss of DNA and no apparent interruption of virulence genes. This second report of chloramphenicol-resistant meningococci is in a setting with a very low volume of systemic chloramphenicol use, but where the high topical use may contribute to recombination events in vivo. Methods for screening for chloramphenicol resistance in meningococci and the in vitro parameters that define this resistance are ill defined. PMID- 14563895 TI - Antiretroviral therapy in sub-Saharan Africa: myth or reality? PMID- 14563896 TI - Meeting the immense need for HAART in resource-poor settings. PMID- 14563897 TI - HIV susceptibility to amprenavir: phenotype-based versus rules-based interpretations. AB - OBJECTIVES: The objective was to study genotypic correlates of discordant interpretations of amprenavir (APV) resistance between a rules-based algorithm and either recombinant phenotype or virtual phenotype. METHODS: HIV resistance mutations found in patients from the GenPheRex study were interpreted with VGI TRUGENE (version 5.0; VGI) and compared with either recombinant-phenotype (Antivirogram, r-PHT) or virtual-phenotype (Virtual-Phenotype, v-PHT) interpreted through Virco biological cut-offs. RESULTS: Among 180 samples available, 56 (31.1%) were discordant with the observed genotype interpretation results, as a result of being judged as sensitive by r-PHT or v-PHT but resistant by VGI (S/R). Only the I84V mutation was almost invariably found in concordant resistant isolates compared with S/R isolates (60% versus 0%, respectively; P < 0.0001). Notwithstanding this, the number of multi-protease inhibitor-associated mutations (PAMs) was significantly higher in the concordant resistant isolates; the prevalence of >3 PAMs was 56.52% versus 33.93% in R/R and S/R isolates, respectively (P = 0.01). Correspondence analysis confirmed the relevance of PAMs, although additional mutations appeared to be correlated with APV resistance. CONCLUSIONS: The rate of discordance between rules-based and either r-PHT or v PHT interpretations for APV was high. Mutation I84V and accumulation of >3 PAMs were found to be associated with resistance as interpreted with all systems tested. However, our results indicate that a number of mutations may have an impact on APV resistance, but that they are missed by current interpretation algorithms and this merits further investigations. PMID- 14563898 TI - Antibacterial susceptibility of a vancomycin-resistant Staphylococcus aureus strain isolated at the Hershey Medical Center. AB - Staphylococcus aureus strain HMC3 isolated at the Hershey Medical Center, was resistant to vancomycin (VRSA) through the presence of the vanA resistance gene; it also contained mecA, erm(A), erm(B), tet(K) and aac(6')-aph(2"), conferring resistance to licensed beta-lactams, macrolides, tetracycline and aminoglycosides. HMC3 also had alterations in GyrA and GrlB and was resistant to available quinolones. Experimental drugs with low MICs (<2 mg/L) for VRSA HMC3 included cephalosporins BAL9141 and RWJ-54428; glycopeptides oritavancin and dalbavancin; the lipopeptide daptomycin; the glycolipodepsipeptide ramoplanin; new fluoroquinolones WCK 771 A, WCK 1153, DK-507k and sitafloxacin; and the DNA nanobinder GS02-02. These agents were all bactericidal as were trimethoprim/sulfamethoxazole and teicoplanin (MIC 4 mg/L). Oxazolidinones linezolid and ranbezolid; the injectable streptogramin quinupristin/dalfopristin; DNA nanobinders GS2-10547 and GS02-104; peptide deformylase inhibitors NVP-PDF713 and GS02-12; tetracycline derivative tigecycline; the antifolate iclaprim; mupirocin and fusidic acid were all active in vitro but bacteriostatic. PMID- 14563899 TI - Oral pharmacokinetically enhanced co-amoxiclav 2000/125 mg, twice daily, compared with co-amoxiclav 875/125 mg, three times daily, in the treatment of community acquired pneumonia in European adults. AB - OBJECTIVES: Pharmacokinetically enhanced co-amoxiclav 2000/125 mg was designed to achieve high serum concentrations of amoxicillin over the 12 h dosing interval to eradicate Streptococcus pneumoniae with amoxicillin MICs of at least 4 mg/L. METHODS: This randomized, double-blind, double-dummy, multicentre study compared the efficacy and safety of oral co-amoxiclav 2000/125 mg twice daily versus co amoxiclav 875/125 mg three times daily, for 7 or 10 days, in the treatment of community-acquired pneumonia (CAP). RESULTS: The per-protocol (PP) population at follow-up (Days 18-39) comprised 114 patients receiving co-amoxiclav 2000/125 mg and 116 receiving co-amoxiclav 875/125 mg. Clinical success at follow-up (primary efficacy endpoint) in the clinical PP population was 94.7% (108/114) for co amoxiclav 2000/125 mg versus 88.8% (103/116) for co-amoxiclav 875/125 mg [treatment difference (TD) = 5.9%, 95% CI: 1.1, 13.0]. Bacteriological success in the bacteriology PP population at follow-up was 85.0% (17/20) for co-amoxiclav 2000/125 mg versus 77.3% (17/22) for co-amoxiclav 875/125 mg (TD = 7.7%, 95% CI: 15.8, 31.2). Penicillin-resistant S. pneumoniae (PRSP) were isolated in three patients (including two with bacteraemia) in the co-amoxiclav 2000/125 mg group (amoxicillin MICs 8 mg/L, penicillin MICs 4 mg/L) and one in the comparator group; all were clinical and bacteriological successes. Co-amoxiclav 2000/125 mg and co-amoxiclav 875/125 mg were associated with adverse events leading to withdrawal in 6.3% and 6.2% of patients, respectively. CONCLUSIONS: Co-amoxiclav 2000/125 mg twice daily was at least as effective clinically as co-amoxiclav 875/125 mg three times daily in the treatment of CAP. Although few patients in this study had PRSP infection, 3/3 were successfully treated with co-amoxiclav 2000/125 mg. PMID- 14563900 TI - Outcomes of an intervention to improve hospital antibiotic prescribing: interrupted time series with segmented regression analysis. AB - OBJECTIVES: To evaluate an intervention to reduce inappropriate use of key antibiotics with interrupted time series analysis. METHODS: The intervention is a policy for appropriate use of Alert Antibiotics (carbapenems, glycopeptides, amphotericin, ciprofloxacin, linezolid, piperacillin-tazobactam and third generation cephalosporins) implemented through concurrent, patient-specific feedback by clinical pharmacists. Statistical significance and effect size were calculated by segmented regression analysis of interrupted time series of drug use and cost for 2 years before and after the intervention started. RESULTS: Use of Alert Antibiotics increased before the intervention started but decreased steadily for 2 years thereafter. The changes in slope of the time series were 0.27 defined daily doses/100 bed-days per month (95% CI 0.19-0.34) and pound 1908 per month (95% CI pound 1238- pound 2578). The cost of development, dissemination and implementation of the intervention ( pound 20133) was well below the most conservative estimate of the reduction in cost ( pound 133296), which is the lower 95% CI of effect size assuming that cost would not have continued to increase without the intervention. However, if use had continued to increase, the difference between predicted and actual cost of Alert Antibiotics was pound 572448 (95% CI pound 435696- pound 709176) over the 24 months after the intervention started. CONCLUSIONS: Segmented regression analysis of pharmacy stock data is a simple, practical and robust method for measuring the impact of interventions to change prescribing. The Alert Antibiotic Monitoring intervention was associated with significant decreases in total use and cost in the 2 years after the programme was implemented. In our hospital, the value of the data far exceeded the cost of processing and analysis. PMID- 14563901 TI - Room for improvement: a systematic review of the quality of evaluations of interventions to improve hospital antibiotic prescribing. AB - INTRODUCTION: In 1999, the British Society for Antimicrobial Chemotherapy (BSAC) and Hospital Infection Society (HIS) convened a working party on optimization of antibiotic prescribing in hospitals. This study was undertaken in order to evaluate the current evidence base on the effectiveness of interventions to change antibiotic prescribing to hospital inpatients. METHODS: We have systematically reviewed the literature from 1980 to identify interventions that alone, or in combination, are effective in improving antibiotic prescribing to hospital inpatients. The protocol was peer reviewed and has been published by the Effective Practice and Organization of Care (EPOC) Group of the Cochrane Collaboration (www.update-software.com/cochrane/). RESULTS: We identified 306 papers, of which 91 (30%) met the minimum inclusion criteria for a Cochrane EPOC review. The reasons for exclusion were uncontrolled before and after design (141/306; 46%) and inadequate interrupted time series (74/306; 24%) with fewer than three observations before and after the intervention. Most of the rejected interrupted time series (ITS) studies had only one or two data points before the intervention with many (up to 15) after it. Only 15 (40%) of the 38 included ITS studies had a statistical analysis and 11 of these used inappropriate statistical tests (e.g. t-test of pre- and post-intervention mean data) rather than analysis of time trends. Regression analysis of the proportion of included studies by year of publication did show a significant positive correlation (R2 = 0.7886). Nonetheless, of 47 papers published since 2000, only 19 (40%) met the minimum eligibility criteria. CONCLUSIONS: The majority of evaluations used fundamentally flawed methodology. There is limited evidence of improvement over time. These problems could be resolved if researchers and referees of protocols or manuscripts implemented the EPOC methodology. PMID- 14563902 TI - Proximal great vessels of aortic arch: comparison of three-dimensional gadolinium enhanced MR angiography and digital subtraction angiography. AB - PURPOSE: To prospectively compare dynamic three-dimensional (3D) gadolinium enhanced magnetic resonance (MR) angiography and digital subtraction angiography (DSA) for the detection of ostial stenosis of the craniocervical vessels. MATERIALS AND METHODS: Thirty-three patients with carotid stenosis of more than 50% at sonography prospectively underwent both MR angiography and DSA. The overall quality of each DSA and MR angiographic study was analyzed. For each craniocervical vessel (brachiocephalic, common carotid, subclavian, and vertebral arteries) (n = 231), ostial stenosis was graded as follows: normal, mild (<50%), moderate to severe (>50%), or occlusion. MR angiographic and DSA results were compared by means of the Spearman rank correlation coefficient (Rs). RESULTS: The overall diagnostic quality of MR angiography was excellent or adequate. Three studies were inadequate because of a poor signal-to-noise ratio (13 of 231 arteries) or a coverage error (five of 231 arteries). Findings at MR angiography and DSA agreed on the degree of stenosis (Rs = 0.82, P <.001). No cases of stenosis of more than 50% were missed at MR angiography. However, some discrepancies were noted between vertebral arteries and the other craniocervical vessels. The sensitivity and specificity for stenosis of more than 50% in other craniocervical vessels were 100% and 98%, respectively. The sensitivity and specificity for stenosis of more than 50% in the vertebral arteries were 100% and 85%, respectively. Findings at MR angiography tended to result in overestimation of the degree of ostial stenosis, especially in vertebral arteries (10 [15%] of 66 arteries). CONCLUSION: MR angiography is useful to rule out ostial stenosis of the craniocervical vessels. MR angiography is an adequate diagnostic tool for ostial stenosis, except in the vertebral artery. PMID- 14563903 TI - Differential diagnosis of hepatic tumors by using contrast enhancement patterns at US. AB - PURPOSE: To assess the accuracy of pattern-based classification of contrast material-enhanced wideband harmonic gray-scale ultrasonographic (US) images in the differential diagnosis of hepatic tumors. MATERIALS AND METHODS: A total of 183 hepatic lesions in 183 patients were studied; lesions included 116 hepatocellular carcinomas, 42 liver metastases, and 25 liver hemangiomas. After injection of a galactose-palmitic acid contrast agent, lesions were scanned with contrast-enhanced wideband harmonic gray-scale US in the arterial, portal venous, and late venous phases. The enhancement patterns were classified, and multiple logistic regression analysis was used to identify diagnostic patterns that enabled differentiation between hepatic tumors. RESULTS: Five enhancement patterns were found to be significant in predicting different hepatic tumors. In hepatocellular carcinomas, the presence of intratumoral vessels in the arterial phase and homogeneous or heterogeneous enhancement in the portal phase were the most typical patterns. In metastases, the absence of intratumoral vessels in the arterial phase and ring enhancement or a perfusion defect in the portal phase were the most typical patterns. In hemangiomas, the absence of intratumoral vessels in the arterial phase and peripheral nodular enhancement in the portal phase were the most typical patterns. The sensitivity, specificity, and accuracy of diagnosis based on combinations of enhancement patterns were, respectively, 94.8%, 94.0%, and 94.5% for hepatocellular carcinoma; 90.5%, 94.3%, and 93.4% for metastasis; and 88.0%, 99.4%, and 97.8% for hemangioma. CONCLUSION: Contrast enhanced wideband harmonic gray-scale US is a useful tool for differentiating among the hepatic tumors studied. PMID- 14563904 TI - Multiplanar and three-dimensional multi-detector row CT of thoracic vessels and airways in the pediatric population. AB - Multi-detector row computed tomography (CT) has changed the approach to imaging of thoracic anatomy and disease in the pediatric population. At the author's institution, multi-detector row CT with multiplanar and three-dimensional reconstruction has become an important examination in the evaluation of systemic and pulmonary vasculature and the tracheobronchial tree. In some clinical situations, multi-detector row CT with reformatted images is obviating conventional angiography, which is associated with higher radiation doses and longer sedation times. Although multi-detector row CT with multiplanar and three dimensional reconstruction is expanding the applications of CT of the thorax, its role as a diagnostic tool still needs to be better defined. The purposes of this article are to describe how to perform multi-detector row CT with multiplanar and three-dimensional reconstruction in young patients, to discuss various reconstruction techniques available, and to discuss applications in the evaluation of vascular and airways diseases. PMID- 14563905 TI - Addressing the potential risks associated with ephedra use: a review of recent efforts. AB - The appropriate amount of oversight for dietary supplements has been a subject of debate for over a decade. This debate has come to a head recently with herbal ephedra, which may be associated with adverse events including heart attack, stroke, seizure, and death. This article reviews and puts into context recent findings on the safety concerns related to ephedra, based primarily on adverse event reports. It presents the response from industry and the FDA in light of this evidence, and describes additional steps taken by other groups who believe that more restrictive action is required. The article concludes by observing the lack of explicit, shared criteria for determining whether a supplement is unsafe, and pointing out ways in which the experience with ephedra can be used constructively to address that problem. PMID- 14563906 TI - The public health response to the Chi-Chi earthquake in Taiwan, 1999. AB - OBJECTIVE: On September 21, 1999, at 1:47 a.m., an earthquake measuring 7.3 on the Richter scale struck the middle Chi-Chi region of Taiwan. The present study examines the response of the public health sector to the earthquake. METHODS: A community needs assessment using modified cluster sampling was performed in shelters of Nantou and Taichung Counties five days after the earthquake struck. Twenty-five temporary medical service systems (TMSSs) conducted surveillance for selected diseases and mortality within one week post-earthquake aided by a buddy system that allowed unaffected counties to provide support to affected counties. RESULTS: The number of cases of acute respiratory infections and acute gastroenteritis in the affected area was higher than that of neighboring unaffected counties in the post-earthquake phase (p<0.001). Earthquake-related deaths were estimated at 2,347 deaths (death rate 116 per 100,000 population); the mean age of the decedents was 49.7 years. No significant difference was observed between males (120/100,000) and females (110/100,000) (risk ratio [RR]=1.09; 95% confidence interval [CI] 0.84, 1.42; p>0.05). The age-adjusted mortality rate was significantly higher in 1999 (odds ratio [OR]=2.11; 95% CI 1.99, 2.24) than in a comparable period in 1998. CONCLUSION: Emergency preparedness must be based on carefully conceived priorities, information, and communications, and improved capabilities must be developed to rapidly implement an emergency public health network. The emergency response to this event consisting of TMSSs, a buddy system, and a communication system-should be considered in planning for future disaster events in Taiwan. PMID- 14563907 TI - Assessment of tuberculosis screening and management practices of large jail systems. AB - OBJECTIVE: This descriptive study sought to explore the use and timeliness of tuberculosis (TB) screening and management activities in jail facilities. METHODS: Study personnel visited 20 large U.S. jail systems and reviewed the medical records of 56 inmates who had recently been evaluated for TB disease and 376 inmates who were diagnosed with or confirmed to have latent TB infection (LTBI). Data from these records were analyzed to determine completion and timeliness of screening, diagnostic, and treatment activities. RESULTS: In 14% of 56 inmates evaluated for TB disease and 24% of 376 inmates with LTBI, chest radiographs were either not performed or not documented. Of 48 inmates evaluated for TB disease who were not receiving treatment when admitted to jail, 10 had no record of sputum collection being done. A mean delay of 3.1 days occurred from symptom report to respiratory isolation. Time from tuberculin skin test reading to chest radiograph reading was a mean of 5.3 days in inmates evaluated for TB disease and a mean of 7.0 days in inmates with LTBI. Follow-up was arranged for 91% of released inmates who were on treatment for TB disease and only 17% of released inmates who were on treatment for LTBI. CONCLUSIONS: Jail health information systems should be augmented to better document and monitor inmate health care related to TB. Completion rates and timeliness of TB screening, diagnostic, and treatment measures should be evaluated to identify areas needing improvement. Finally, mechanisms for continuity of care upon inmate release should be enhanced to promote therapy completion and prevent TB transmission in the community. PMID- 14563908 TI - Content and criterion validity evaluation of National Public Health Performance Standards measurement instruments. AB - OBJECTIVE: The Centers for Disease Control and Prevention's National Public Health Performance Standards Program (NPHPSP) has developed instruments to measure the performance of local and state public health departments on the 10 "Essential Services of Public Health," which have been tested in several states. This article is a report of the evaluation of the content and criterion validity of the local public health performance assessment instrument, and the content validity of the state public health performance assessment instrument. METHODS: Health department performance is measured using a set of indicators developed for the 10 Essential Services of Public Health and a model standard for each indicator. Content validity of each model standard in the local instrument was addressed by community partners along the following dimensions: the importance of each standard as a measure of the associated Essential Service, its completeness as a measure, and its reasonableness for achievement. All standards for each Essential Service were then judged in terms of their completeness in measuring performance in that service. Content validity of the state instrument was evaluated in a group interview of health department staff members from three states. Criterion validity of the local instrument was assessed for a sample of eight public health departments in Florida and six in New York by examining documentary evidence for selected responses. Criterion validity was also evaluated for a sample of Florida local public health departments and one Hawaii public health department by comparing state health department staffs' judgments of performance against the instrument score. RESULTS: Criterion validity was upheld for a summary performance score on the local instrument, but was not upheld for performance judgments on individual Essential Services. The NPHPSP standards based on the Essential Services have validity for measuring local public health system performance, according to community partners. The model standards are valid measures of state performance, according to state public health departments in three states. CONCLUSIONS: Within the scope of the validity evaluations completed, the NPHPSP state and local performance assessment instruments were found to be valid measures of public health performance. PMID- 14563909 TI - Alaska native mortality, 1979-1998. AB - OBJECTIVES: This study compares mortality patterns for the Alaska Native population and the U.S. white population for 1989-1998 and examines trends for the 20-year period 1979-1998. METHODS: The authors used death certificate data and Indian Health Service population estimates to calculate mortality rates for the Alaska Native population, age-adjusted to the U.S. 1940 standard million. Data on population and mortality for U.S. whites, aggregated by 10-year age groups and by gender, were obtained from the National Center for Health Statistics, and U.S. white mortality rates were age-adjusted to the U.S. 1940 standard million. RESULTS: Overall, 1989-1998 Alaska Native mortality rates were 60% higher than those for the U.S. white population for the same period. There were significant disparities for eight of 10 leading causes of death, particularly unintentional injury, suicide, and homicide/legal intervention. Although declines in injury rates can be documented for the period 1979-1998, large disparities still exist. Alaska Native death rates for cancer, cerebrovascular disease, chronic obstructive pulmonary disease, and diabetes increased from 1979 to 1998. Given decreases in some cause-specific mortality rates in the U.S. white population, increased rates among Alaska Natives have resulted in new disparities. CONCLUSIONS: These data indicate that improvements in injury mortality rates are offset by marked increases in chronic disease deaths. PMID- 14563910 TI - Alternative HIV testing methods among populations at high risk for HIV infection. AB - OBJECTIVE: The purpose of this study was to determine the levels of awareness and use of alternative HIV tests (home collection kit, oral mucosal transudate collection kit, and rapid tests) among people at high risk for HIV infection. METHODS: Data were collected as part of an anonymous, cross-sectional interview study--the HIV Testing Survey (HITS)--conducted in seven states from September 2000 to February 2001. Three high-risk populations were recruited: men who have sex with men, injection drug users, and high-risk heterosexuals. Respondents were asked about their awareness and use of alternative HIV tests. RESULTS: The overall awareness and use of the alternative tests was limited: 54% of respondents were aware of the home collection kit, 42% were aware of the oral mucosal transudate collection kit test, and 13% were aware of rapid tests. Among those aware of alternative tests, self-reported use of the tests was also low. The most common reasons given for not using alternative HIV tests were: preference for the standard test; concern that the results could be less accurate; and that alternative tests were not offered. CONCLUSIONS: The low levels of awareness and use of alternative HIV tests suggest that the potential for promoting testing among individuals at high risk for HIV by encouraging use of alternative HIV tests has not been fully realized. Alternative tests should be made more broadly available and should be accompanied by education about these tests for physicians and people at risk. Educational efforts should be evaluated to determine if promoting alternative HIV tests increases the numbers of people at risk for HIV who are tested. PMID- 14563911 TI - Correlates of sunburn experiences among U.S. adults: results of the 2000 National Health Interview Survey. AB - OBJECTIVE: The purpose of this study was to determine the rate of sunburns in the U.S. adult population and the correlates of sunburns. METHODS: Data from the 2000 National Health Interview Survey Cancer Control Module were used to calculate the number of sunburns (0, 1, 2, or > or = 3) experienced during the past year by age, sex, race/ethnicity, and skin sensitivity to sun exposure. The relationship between no sunburns vs. one or more sunburns and additional demographic, health, and behavioral factors for adults who self-identify as white Hispanic or white non-Hispanic was assessed using general linear contrasts. Multivariate logistic regression modeling was conducted to determine the most important covariates associated with sunburns. All analyses were weighted for the complex sampling design. RESULTS: The study data suggest that overall, 18.5% (95% confidence interval [CI] 17.9, 19.1) of U.S. adults experience one sunburn a year, 9.7% (95% CI 9.3, 10.1) experience two, and 8.0% (95% CI 7.6, 8.4) experience > or = 3 sunburns. The data also indicate that adults who self-identify as white non Hispanic experience sunburns more frequently than (in order of prevalence) those who identify as American Indian/Alaska Native, white Hispanic, Asian/Pacific Islander, or black. Sunburns were found to be more common among men than among women, more common among younger age groups than among older age groups, and more common among those with skin more prone to sunburn than among those with skin less prone to sunburn. Among individuals who self-identify as white Hispanic or white non-Hispanic, protective behaviors associated with lower rates of one or more sunburns in multivariate analyses are staying in the shade (odds ratio [OR] 0.73, 95% CI 0.66, 0.80) and wearing long-sleeved shirts (OR 0.86, 95% CI 0.75, 0.99). CONCLUSIONS: Many American adults have one or more sunburns per year. Methods to protect from sun exposure may not be used as needed to prevent sunburn. PMID- 14563913 TI - Lawrence v Texas: implications for public health policy and practice. PMID- 14563912 TI - An economic assessment of pre-vaccination screening for hepatitis A and B. AB - OBJECTIVE: The availability of a single vaccine active against hepatitis A and B may facilitate prevention of both infections, but complicates the question of whether to conduct pre-vaccination screening. The authors examined the cost effectiveness of pre-vaccination screening for several populations: first-year college students, military recruits, travelers to hepatitis A-endemic areas, patients at sexually transmitted disease clinics, and prison inmates. METHODS: Three prevention protocols were examined: (1) screen and defer vaccination until serology results are known; (2) screen and begin vaccination immediately to avoid a missed vaccination opportunity; and (3) vaccinate without screening. Data describing pre-vaccination immunity, vaccine effectiveness, and prevention costs borne by the health system (i.e., serology, vaccine acquisition, and administration) were derived from published literature and U.S. government websites. Using spreadsheet models, the authors calculated the ratio of prevention costs to the number of vaccine protections conferred. RESULTS: The vaccinate without screening protocol was most cost-effective in nine of 10 analyses conducted under baseline assumptions, and in 69 of 80 sensitivity analyses. In each population considered, vaccinate without screening was less costly than and at least equally as effective as screen and begin vaccination. The screen and defer vaccination protocol would reduce costs in seven populations, but effectiveness would also be lower. CONCLUSIONS: Unless directed at vaccination candidates with the highest probability of immunity, pre vaccination screening for hepatitis A and B immunity is not cost-effective. Balancing cost reduction with reduced effectiveness, screen and defer may be preferred for older travelers and prison inmates. PMID- 14563916 TI - Evaluation of the effectiveness of the South Central Center for Public Health Preparedness training. PMID- 14563919 TI - Transabdominal near infrared oximetry of hypoxic stress in fetal sheep brain in utero. AB - The feasibility of transabdominal near-infrared (NIR) spectroscopy for detecting and quantifying fetal hypoxia in utero is demonstrated in a pregnant ewe model. A frequency domain NIR spectroscopy probe, consisting of two detectors and six sources operating at three wavelengths (675, 786, and 830 nm), was placed on the maternal abdomen directly above the fetal head. Fetal hypoxia was indirectly induced through occlusion of uterine blood flow for approximately 3 min. NIR photon diffusion measurements were made during a baseline period, during hypoxia of the fetus, and during recovery. Fetal blood samples were drawn from the fetal brachial artery and jugular veins at several time points during the cycle. Seven hypoxic cycles were induced in a total of five pregnant ewes. The NIR measurements were analyzed by using a two-layer diffusion model to deconvolve the fetal blood saturation from that of the pregnant ewe. Fetal hypoxia was detected. Good agreement was found between fetal blood saturation determined by the transabdominal NIR method and arterial and venous fetal blood saturation quantified from fetal blood samples by using a hemoximeter. PMID- 14563920 TI - Localization and roles of Ski8p protein in Sordaria meiosis and delineation of three mechanistically distinct steps of meiotic homolog juxtaposition. AB - Ski8p is implicated in degradation of non-poly(A) and double-stranded RNA, and in meiotic DNA recombination. We have identified the Sordaria macrospora SKI8 gene. Ski8p is cytoplasmically localized in all vegetative and sexual cycle cells, and is nuclear localized, specifically in early-mid-meiotic prophase, in temporal correlation with Spo11p, the meiotic double-strand break (DSB) transesterase. Localizations of Ski8p and Spo11p are mutually interdependent. ski8 mutants exhibit defects in vegetative growth, entry into the sexual program, and sporulation. Diverse meiotic defects, also seen in spo11 mutants, are diagnostic of DSB absence, and they are restored by exogenous DSBs. These results suggest that Ski8p promotes meiotic DSB formation by acting directly within meiotic prophase chromosomes. Mutant phenotypes also divide meiotic homolog juxtaposition into three successive, mechanistically distinct steps; recognition, presynaptic alignment, and synapsis, which are distinguished by their differential dependence on DSBs. PMID- 14563921 TI - Eradication of pathogenic beta-catenin by Skp1/Cullin/F box ubiquitination machinery. AB - The use of Skp1/Cull 1/F box (SCF) ubiquitin-conjugation machinery as a potential knockout tool offers a means of eradicating disease-causing proteins. Here a chimeric F box protein (CFP) was engineered to achieve selective eradication of pathogenic beta-catenin in colorectal cancer. We show that CFP specifically searches for and subsequently links the abnormal beta-catenin to the cellular SCF ubiquitination complex. Introduction of the CFP to colorectal cancer cells induced targeted ubiquitination and proteolytic degradation of nuclear and cytoplasmic free beta-catenin while preserving its normal cellular adhesion counterpart. Elimination of pathogenic beta-catenin suppressed constitutive Wingless/Wnt signaling and inhibited in vitro and in vivo tumor cell growth. This study demonstrates a practical utility of a SCF-based knockout system as a tool in targeting an abnormal protein that affects growth and transformation. PMID- 14563922 TI - Damped elastic recoil of the titin spring in myofibrils of human myocardium. AB - The giant protein titin functions as a molecular spring in muscle and is responsible for most of the passive tension of myocardium. Because the titin spring is extended during diastolic stretch, it will recoil elastically during systole and potentially may influence the overall shortening behavior of cardiac muscle. Here, titin elastic recoil was quantified in single human heart myofibrils by using a high-speed charge-coupled device-line camera and a nanonewtonrange force sensor. Application of a slack-test protocol revealed that the passive shortening velocity (Vp) of nonactivated cardiomyofibrils depends on: (i) initial sarcomere length, (ii) release-step amplitude, and (iii) temperature. Selective digestion of titin, with low doses of trypsin, decelerated myofibrillar passive recoil and eventually stopped it. Selective extraction of actin filaments with a Ca2+-independent gelsolin fragment greatly reduced the dependency of Vp on release-step size and temperature. These results are explained by the presence of viscous forces opposing myofibrillar passive recoil that are caused mainly by weak actin-titin interactions. Thus, Vp is determined by two distinct factors: titin elastic recoil and internal viscous drag forces. The recoil could be modeled as that of a damped entropic spring consisting of independent worm-like chains. The functional importance of myofibrillar elastic recoil was addressed by comparing instantaneous Vp to unloaded shortening velocity, which was measured in demembranated, fully Ca2+-activated, human cardiac fibers. Titin-driven passive recoil was much faster than active unloaded shortening velocity in early phases of isotonic contraction. Damped myofibrillar elastic recoil could help accelerate active contraction speed of human myocardium during early systolic shortening. PMID- 14563923 TI - The small RING finger protein Z drives arenavirus budding: implications for antiviral strategies. AB - By using a reverse genetics system that is based on the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV), we have identified the arenavirus small RING finger Z protein as the main driving force of virus budding. Both LCMV and Lassa fever virus (LFV) Z proteins exhibited self-budding activity, and both substituted efficiently for the late domain that is present in the Gag protein of Rous sarcoma virus. LCMV and LFV Z proteins contain proline-rich motifs that are characteristic of late domains. Mutations in the PPPY motif of LCMV Z severely impaired the formation of virus-like particles. LFV Z contains two different proline-rich motifs, PPPY and PTAP, which are separated by eight amino acids. Mutational analysis revealed that both motifs are required for efficient LFV Z mediated budding. Both LCMV and LFV Z proteins recruited to the plasma membrane Tsg101, which is a component of the class E vacuolar protein sorting machinery that has been implicated in budding of HIV and Ebola virus. Targeting of Tsg101 by RNA interference caused a strong reduction in Z-mediated budding. These results indicate that Z is the arenavirus functional counterpart of the matrix proteins found in other negative strand enveloped RNA viruses. Moreover, members of the vacuolar protein sorting pathway appear to play an important role in arena virus budding. These findings open possibilities for antiviral strategies to combat LFV and other hemorrhagic fever arenaviruses. PMID- 14563925 TI - Ancient maize from Chacoan great houses: where was it grown? AB - In this article, we compare chemical (87Sr/86Sr and elemental) analyses of archaeological maize from dated contexts within Pueblo Bonito, Chaco Canyon, New Mexico, to potential agricultural sites on the periphery of the San Juan Basin. The oldest maize analyzed from Pueblo Bonito probably was grown in an area located 80 km to the west at the base of the Chuska Mountains. The youngest maize came from the San Juan or Animas river floodplains 90 km to the north. This article demonstrates that maize, a dietary staple of southwestern Native Americans, was transported over considerable distances in pre-Columbian times, a finding fundamental to understanding the organization of pre-Columbian southwestern societies. In addition, this article provides support for the hypothesis that major construction events in Chaco Canyon were made possible because maize was brought in to support extra-local labor forces. PMID- 14563924 TI - An open conformation of mammalian cytochrome P450 2B4 at 1.6-A resolution. AB - The xenobiotic metabolizing cytochromes P450 (P450s) are among the most versatile biological catalysts known, but knowledge of the structural basis for their broad substrate specificity has been limited. P450 2B4 has been frequently used as an experimental model for biochemical and biophysical studies of these membrane proteins. A 1.6-A crystal structure of P450 2B4 reveals a large open cleft that extends from the protein surface directly to the heme iron between the alpha helical and beta-sheet domains without perturbing the overall P450 fold. This cleft is primarily formed by helices B' to C and F to G. The conformation of these regions is dramatically different from that of the other structurally defined mammalian P450, 2C5/3LVdH, in which the F to G and B' to C regions encapsulate one side of the active site to produce a closed form of the enzyme. The open conformation of 2B4 is trapped by reversible formation of a homodimer in which the residues between helices F and G of one molecule partially fill the open cleft of a symmetry-related molecule, and an intermolecular coordinate bond occurs between H226 and the heme iron. This dimer is observed both in solution and in the crystal. Differences between the structures of 2C5 and 2B4 suggest that defined regions of xenobiotic metabolizing P450s may adopt a substantial range of energetically accessible conformations without perturbing the overall fold. This conformational flexibility is likely to facilitate substrate access, metabolic versatility, and product egress. PMID- 14563926 TI - The detection and quantitation of free desmosine and isodesmosine in human urine and their peptide-bound forms in sputum. AB - Desmosine (D) and isodesmosine (I), the intramolecular crosslinking amino acids that occur in chains of elastin, have now been found in free form in human urine. Until now, these amino acids (M(r) = 526) were found to occur in urine only as higher molecular weight (M (r) = 1,000-1,500) peptides. Thus, the previously used analytical methods required, as the first step, acid hydrolysis of the urine at elevated temperature to liberate D and I from their peptides. The analytical method described here uses HPLC followed by electrospray ionization MS for the detection and quantitation of free D and I in unhydrolyzed urine. Identities of both D and I were established by their retention times on LC and by their mass ion at 526 atomic mass units, characteristic of each compound. The sensitivity of the method is 0.10 ng. The average values of free D and I in the urine of seven healthy subjects were 1.42 +/- 1.16 and 1.39 +/- 1.04 microg/g of creatinine, respectively. After acid hydrolysis of the urine, the amounts of D and I were 8.67 +/- 3.75 and 6.28+/-2.87 microg/g of creatinine, respectively. The method was also successfully used to measure peptide-bound D and I levels in the sputum of patients with chronic obstructive pulmonary disease. PMID- 14563929 TI - What affects mRNA levels in leaves of field-grown aspen? A study of developmental and environmental influences. AB - We have analyzed the abundance of mRNAs expressed from 11 nuclear genes in leaves of a free-growing aspen (Populus tremula) tree throughout the growing season. We used multivariate statistics to determine the influence of environmental factors (i.e. the weather before sampling) and developmental responses to seasonal changes at the mRNA level for each of these genes. The gene encoding a germin like protein was only expressed early in the season, whereas the other tested genes were expressed throughout the season and showed mRNA variations on a day-to day basis. For six of the genes, reliable models were found that described the mRNA level as a function of weather, but the leaf age was also important for all genes except one encoding an early light-inducible protein (which appeared to be regulated purely by environmental factors under these conditions). The results confirmed the importance of several environmental factors previously shown to regulate the genes, but we also detected a number of less obvious factors (such as the variation in weather parameters and the weather of the previous day) that correlated with the mRNA levels of individual genes. The study shows the power of multivariate statistical methods in analyzing gene regulation under field conditions. PMID- 14563927 TI - Phylogenetic analyses and expression studies reveal two distinct groups of calreticulin isoforms in higher plants. AB - Calreticulin (CRT) is a multifunctional protein mainly localized to the endoplasmic reticulum in eukaryotic cells. Here, we present the first analysis, to our knowledge, of evolutionary diversity and expression profiling among different plant CRT isoforms. Phylogenetic studies and expression analysis show that higher plants contain two distinct groups of CRTs: a CRT1/CRT2 group and a CRT3 group. To corroborate the existence of these isoform groups, we cloned a putative CRT3 ortholog from Brassica rapa. The CRT3 gene appears to be most closely related to the ancestral CRT gene in higher plants. Distinct tissue dependent expression patterns and stress-related regulation were observed for the isoform groups. Furthermore, analysis of posttranslational modifications revealed differences in the glycosylation status among members within the CRT1/CRT2 isoform group. Based on evolutionary relationship, a new nomenclature for plant CRTs is suggested. The presence of two distinct CRT isoform groups, with distinct expression patterns and posttranslational modifications, supports functional specificity among plant CRTs and could account for the multiple functional roles assigned to CRTs. PMID- 14563928 TI - HY5, Circadian Clock-Associated 1, and a cis-element, DET1 dark response element, mediate DET1 regulation of chlorophyll a/b-binding protein 2 expression. AB - DET1 is a pleiotropic regulator of Arabidopsis development and controls the expression of many light-regulated genes. To gain a better understanding of the mechanism by which DET1 controls transcription from light-regulated promoters, we identified elements in the chlorophyll a/b-binding protein 2 (CAB2) promoter that are required for DET1-mediated expression. Using a series of reporter constructs in which the luciferase gene is controlled by CAB2 promoter fragments, we defined two DET1-responsive elements in the CAB2 promoter that are essential for proper CAB2 transcription. A 40-bp DET1 dark-response element (DtRE) is required for both dark and root-specific repression of CAB2, whereas the known CAB upstream factor-1 element is required for DET1 activation-associated effects in the light and repression in the roots. HY5, a factor that binds CAB upstream factor-1, is also required for DET1 effects in the light. DtRE binds two distinct activities in Arabidopsis seedling extracts: a novel activity with binding site CAAAACGC that we have named CAB2 DET1-associated factor 1 plus an activity that is likely to be the myb transcription factor Circadian Clock-Associated 1. Both activities are altered in dark-grown det1 extracts as compared with wild type, correlating a change in extractable DNA binding activity with a major change in CAB2 expression. We conclude that DET1 represses the CAB2 promoter in the dark by regulating the binding of two factors, CAB2 DET1-associated factor 1 and Circadian Clock-Associated 1, to the DtRE. PMID- 14563930 TI - Arabidopsis PSEUDO-RESPONSE REGULATOR7 is a signaling intermediate in phytochrome regulated seedling deetiolation and phasing of the circadian clock. AB - To identify new components in the phytochrome (phy) signaling network in Arabidopsis, we used a sensitized genetic screen for deetiolation-defective seedlings. Two allelic mutants were isolated that exhibited reduced sensitivity to both continuous red and far-red light, suggesting involvement in both phyA and phyB signaling. The molecular lesions responsible for the phenotype were shown to be mutations in the Arabidopsis PSEUDO-RESPONSE REGULATOR7 (PRR7) gene. PRR7 is a member of a small gene family in Arabidopsis previously suggested to be involved in circadian rhythms. A PRR7-beta-glucuronidase fusion protein localized to the nucleus, implying a possible function in the regulation of photoresponsive gene expression. Consistent with this suggestion, prr7 seedlings were partially defective in the regulation of the rapidly light-induced genes CIRCADIAN CLOCK ASSOCIATED1 (CCA1) and LATE ELONGATED HYPOCOTYL (LHY), observable as a premature increase in expression level during the second peak of the biphasic induction profile that is elicited upon initial exposure of dark-grown seedlings to light. A similar 3- to 6-h coordinated advance in peak free-running expression of CCA1, LHY, and TIMING-OF-CAB1, which are considered to encode the molecular components of the circadian oscillator in Arabidopsis, was observed in entrained fully green prr7 seedlings compared with wild-type seedlings. Collectively, these data suggest that PRR7 functions as a signaling intermediate in the phytochrome regulated gene expression responsible for both seedling deetiolation and phasing of the circadian clock in response to light. PMID- 14563933 TI - Imaging analysis of subcellular correlation of androgen receptor and estrogen receptor alpha in single living cells using green fluorescent protein color variants. AB - Androgen and estrogen act not only in a sex-specific manner but also interactively and synergistically. In the present study, to examine the possible interaction between androgen receptor (AR) and estrogen receptor-alpha (ERalpha), we investigated the subcellular dynamics of AR and ERalpha fused with green fluorescent protein color variants in single living cells using time-lapse microscopy and the technique of fluorescence recovery after photobleaching. AR and ERalpha showed punctate colocalization in the nucleus with estrogen, but not androgen. N-terminal AR deletion mutant did not form a nuclear punctate pattern with either androgen or estrogen. In the presence of AR, but not ERalpha, N terminal AR deletion mutant formed a punctate nuclear pattern with androgen. AR had different mobility depending on the ligand and the presence of ERalpha. On the other hand, AR had little effect on the stability of ERalpha. ERalpha mutant that does not bind coactivators did not alter the mobility of AR. Taken together, using an imaging technique, we clarified that possible homo/hetero dimerization between AR and ERalpha could be attributed to androgen-estrogen interaction in living cells. PMID- 14563931 TI - DETERMINATE and LATE FLOWERING are two TERMINAL FLOWER1/CENTRORADIALIS homologs that control two distinct phases of flowering initiation and development in pea. AB - Genes in the TERMINAL FLOWER1 (TFL1)/CENTRORADIALIS family are important key regulatory genes involved in the control of flowering time and floral architecture in several different plant species. To understand the functions of TFL1 homologs in pea, we isolated three TFL1 homologs, which we have designated PsTFL1a, PsTFL1b, and PsTFL1c. By genetic mapping and sequencing of mutant alleles, we demonstrate that PsTFL1a corresponds to the DETERMINATE (DET) gene and PsTFL1c corresponds to the LATE FLOWERING (LF) gene. DET acts to maintain the indeterminacy of the apical meristem during flowering, and consistent with this role, DET expression is limited to the shoot apex after floral initiation. LF delays the induction of flowering by lengthening the vegetative phase, and allelic variation at the LF locus is an important component of natural variation for flowering time in pea. The most severe class of alleles flowers early and carries either a deletion of the entire PsTFL1c gene or an amino acid substitution. Other natural and induced alleles for LF, with an intermediate flowering time phenotype, present no changes in the PsTFL1c amino acid sequence but affect LF transcript level in the shoot apex: low LF transcript levels are correlated with early flowering, and high LF transcript levels are correlated with late flowering. Thus, different TFL1 homologs control two distinct aspects of plant development in pea, whereas a single gene, TFL1, performs both functions in Arabidopsis. These results show that different species have evolved different strategies to control key developmental transitions and also that the genetic basis for natural variation in flowering time may differ among plant species. PMID- 14563934 TI - A silencer element in the first intron of the glutamine synthetase gene represses induction by glucocorticoids. AB - The enzyme glutamine synthetase (GS) ranks as one of the most remarkable glucocorticoid-inducible mammalian genes. In many tissues and cell lines, the synthetic glucocorticoid dexamethasone alone increases GS expression several fold. The direct response is mainly mediated by a cellular glucocorticoid receptor that, upon binding of the hormone, interacts with glucocorticoid responsive elements (GREs) of the gene. In cells of hepatocellular origin the response is mediated by a GRE located in the first intron of the gene. Surprisingly, hepatocytes do not respond to glucocorticoids with enhanced GS expression, despite the presence of an intact glucocorticoid receptor, which, in the same cells, stimulates expression of other genes such as tyrosine amino transferase. Reporter gene assays identified a sequence element downstream from the intronic GRE that inhibits the enhancement of expression by glucocorticoids. This silencer was designated GS silencer element of the rat. Gel mobility shift assays demonstrate the binding of a factor in hepatocyte nuclear extract. This yet unknown factor was designated GS silencer-binding protein. It is absent in FAO cells that respond to glucocorticoids with enhanced expression of GS and present in HepG2 cells that do not respond. PMID- 14563935 TI - The role of follistatin domains in follistatin biological action. AB - Follistatin (FS) is an important regulator of pituitary FSH secretion through its potent ability to bind and bioneutralize activin. It also represents a prototype for binding proteins that control bioavailability of other TGFbeta-related growth factors such as the bone morphogenetic proteins. The 288-residue FS molecule has a distinctive structure comprised principally of three 10-cysteine FS domains. These are preceded by an N-terminal segment shown by us previously to contain hydrophobic residues essential for activin binding. To establish the contribution of the FS domains themselves to FS's bioactivity, we prepared mutants with deleted or exchanged domains and intradomain point mutations. Mutants were expressed from mammalian (Chinese hamster ovary) cells and evaluated for activin binding and for biological activity in assays measuring differing aspects of FS bioactivity: activin-mediated transcriptional activity and suppression of FSH secretion in primary pituitary cell cultures. The N-terminal domain (residues 1 63) alone could not bind activin or suppress activin-mediated transcription, either alone or combined in solution with the FS domain region (residues 64-288). Deletion of FS domains 1 or 2 abolished activin binding and biological activity in both assays, whereas deletion of domain 3 was tolerated. Bioactivity was also reduced or eliminated after exchange of domains (FS 2/1/3 and FS 3/1/2) or doubling of domain 1 (FS 1/1/3) or domain 2 (FS 2/2/3). Several hydrophobic residues clustered within the C-terminal region of FS domains 1 and 2 are highly conserved among all FS domains. Mutation of any of these to Asp or Ala either reduced or eliminated FS bioactivity and disrupted distant epitopes for heparin binding (FS domain 1) or antibody recognition (FS domain 2), suggesting their role in maintaining the conformational integrity of the domain and possibly the FS molecule as a whole. These results are consistent with the importance of domain conformation as well as the overall order of the domains in FS function. A continuous sequence comprising the N-terminal domain and followed by FS domains 1 and 2 fulfills the minimum structural requirement for activin binding and FS bioactivity. PMID- 14563936 TI - Gene expression profile in thyroid of transgenic mice overexpressing the adenosine receptor 2a. AB - Mutations of the TSH receptor leading to constitutive activation of the cAMP cascade are responsible for the development of hot nodules, if arising in a somatic cell, and nonautoimmune hyperthyroidism, when occurring in a germinal cell. An animal model of constitutive activation of the thyroid cAMP cascade has been obtained by generating transgenic mice expressing the adenosine receptor (Tg A2aR) under the control of the thyroglobulin promoter. These mice develop huge goiters and die prematurely due to hyperthyroidism induced cardiac failure. To identify new genes involved in the tumorigenic pathway of the thyroid, we designed a protocol using microarray technology to study the differential expression, between normal and transgenic thyroid, of +/-13,000 genes. A total of 360 genes or expressed sequence tags showed a strong modulation with background corrected values of fluorescence superior to 2-fold change. The modulated genes were classified according to their proposed gene ontology functions. Approximately half of them were up-regulated. The function of the majority of these genes in thyroid physiology is still to be determined. Some of them, like IGF-I or IGF binding protein 3 or 5, may play an important role in the development of thyroid nodules through paracrine mechanisms. This study demonstrates the feasibility of sequentially following the cascade of events leading to the formation of benign tumors such as hot thyroid nodule or hyperfunctional goiter. PMID- 14563937 TI - Fibroblast growth factor receptor 3 gene: regulation by serum response factor. AB - We have previously identified a cis-acting sequence in the proximal promoter of the fibroblast growth factor receptor 3 (FGFR3) gene that strongly activates transcription in chondrocytic cells. Here we report that the transcriptional activity of this sequence (FRE3) requires serum response factor and its cognate recognition motif, serum response element. Although the FRE3 contains consensus sequence motifs for several transcription factors, the serum response element is paramount for the transcriptional activity of the FRE3. Additionally, the transcriptional activity of the proximal promoter of the FGFR3 gene is suppressed by mutation of the serum response element. Serum response factor binds to the FRE3 as evidenced by gel shift experiments and antibody supershift experiments and expression of a dominant negative form of serum response factor suppresses the activity of FRE3. Additionally, serum response factor binds to the FGFR3 gene in vivo, as demonstrated by chromatin immunoprecipitation. Serum response factor is an important regulator of cardiac, skeletal, and smooth muscle gene expression; these data suggest that serum response factor is also an important determinant of chondrocyte gene expression. PMID- 14563938 TI - Making sense of cross-talk between steroid hormone receptors and intracellular signaling pathways: who will have the last word? AB - In classical models of nuclear steroid hormone receptor function, ligand binds receptor, heat shock proteins dissociate, and receptor dimers enter or are withheld in the nucleus and interact with coregulatory molecules to mediate changes in gene expression. The footnotes, "receptors become phosphorylated" and "dynamic nucleo-cytoplasmic shuttling occurs" describe well-accepted, but less well-understood aspects of receptor action. Recently, the idea that several protein kinases are activated in response to steroid hormone binding to cognate cytoplasmic or membrane-associated receptors has become fashionable. However, the precise role of steroid hormone receptor phosphorylation and our understanding of which cytoplasmic kinases are activated and their functional significance remain elusive. This review provides an overview of the primary ways in which steroid hormone receptor and growth factor cross-talk occurs, using the human progesterone receptor (PR) as a model. The functional consequences of PR phosphorylation by protein kinases classically activated in response to peptide growth factors and novel extranuclear or nongenomic functions of PR as potential independent initiators of signal transduction pathways are discussed. Intracellular protein kinases are emerging as key mediators of steroid hormone receptor action. Cross-talk between steroid receptor- and growth factor-initiated signaling events may explain how gene subsets are coordinately regulated by mitogenic stimuli in hormonally responsive normal tissues, and is suspected to play a role in their cancer biology. PMID- 14563941 TI - A crucial role of hepatocyte nuclear factor-4 expression in the differentiation of human ductular hepatocytes. AB - Ductular structures are suggested to be bipotential progenitor cells that may differentiate into hepatocytes or biliary epithelial cells (BEC). To better understand the differentiation process, we studied the expression of hepatocyte nuclear factor (HNF) in ductular structures. Matured hepatocytes in normal liver expressed HNF-1, HNF-4alpha, HNF-3beta, and C/EBPalpha in the nucleus. Normal BEC expressed HNF-1 but did not express HNF-4alpha, suggesting an important role of HNF-4alpha in maintaining the phenotype of matured hepatocytes. Ductular structures were classified into ductular cells and ductular hepatocytes. Ductular cells showed glandular or bile duct-like appearance and strongly expressed cytokeratin-7. Ductular hepatocytes showed features between BEC and hepatocytes and heterogeneously expressed cytokeratin-7. Both ductular cells and ductular hepatocytes expressed HNF-4alpha, but the nuclear localization of HNF-4alpha was more prominent in ductular hepatocytes. The expression of HNF-4alpha mRNA in ductular hepatocytes was demonstrated at the single cell level by laser capture microdissection. Regenerative hepatocytes strongly expressed all HNFs in the nucleus, whereas residual hepatocytes in massive necrosis showed low or cytoplasmic expression. These results suggest that HNF-4alpha plays an important role in the differentiation and maintenance of the matured hepatocyte phenotype and that nuclear localization of HNFs is implicated in the accomplishment of their function. PMID- 14563940 TI - COX-2/VEGF-dependent facilitation of tumor-associated angiogenesis and tumor growth in vivo. AB - Nonsteroidal anti-inflammatory drugs are known to suppress the occurrence and progression of malignancies such as colorectal cancers. However, the precise mechanism of these actions remains unknown. We have evaluated the role of an inducible cyclo-oxygenase (COX-2) in tumor-associated angiogenesis and tumor growth, and identified the downstream molecules involved using a ddy mouse model of sponge angiogenesis, which mimics tumor angiogenesis and is COX-2 and vascular endothelial growth factor (VEGF) dependent. In this model, VEGF expression was down-regulated by selective COX-2 inhibition with NS-398. To find out the involvement of COX-2/VEGF pathway in tumor-associated angiogenesis, we estimated angiogenesis occurring around implanted Millipore chambers containing sarcoma-180 (S-180) cells or Lewis lung carcinoma cells. Daily oral administration of NS-398 or of aspirin, a nonselective COX inhibitor, suppressed angiogenesis seen around the Millipore chambers. S-180 cells implanted in ddy mice formed substantial tumors with extensive angiogenesis markedly suppressed by aspirin and COX-2 inhibitors NS-398 and JTE522, but not by mofezolac, an inhibitor of constitutive COX-1. Tumor-associated angiogenesis was also significantly suppressed by a neutralizing antibody against VEGF. S-180 tumor growth in the subcutaneous tissues was also suppressed by aspirin, COX-2 selective inhibitors, and the VEGF antibody, but not by the COX-1 inhibitor. These results demonstrate that the inhibition of the COX-2/VEGF-dependent pathway was effective in tumor-associated angiogenesis, tumor growth, and tumor metastasis. PMID- 14563942 TI - Aberrant expression of trefoil factor family 1 in biliary epithelium in hepatolithiasis and cholangiocarcinoma. AB - Stepwise progression of intrahepatic cholangiocarcinoma (ICC) has been proposed in hepatolithiasis. We examined the participation of trefoil factor family 1 (TFF1), which is critical for mucosal protection and tumor suppression in the stomach, in the development and progression of ICC. We used 16 livers of ICC with hepatolithiasis, 11 of biliary epithelial dysplasia with hepatolithiasis, 16 of hepatolithiasis without dysplasia or carcinoma, 18 of ICC without hepatolithiasis, and 39 control livers. TFF1 expression in the biliary epithelium was increased in hepatolithiasis compared with control livers (p < 0.01). In biliary epithelial dysplasia and noninvasive ICC with hepatolithiasis, TFF1 was extensively expressed and MUC5AC gastric mucin was usually colocalized with TFF1. However, TFF1 expression was significantly decreased in invasive ICC despite preserved expression of MUC5AC. A total of four missense mutations were detected: three in two noninvasive ICC with hepatolithiasis (28.6%) and one in invasive ICC (11%). Loss of heterozygosity of the TFF1 gene was not detectable. The decreased expression of TFF1 in invasive ICC may be explained by the methylation of the TFF1 promoter region. Up-regulation of TFF1 coupled with MUC5AC in biliary epithelium in hepatolithiasis, biliary epithelial dysplasia, and noninvasive ICC may reflect the gastric metaplasia and early neoplastic lesion. Under such conditions, decreased TFF1 expression may lead to increased cell proliferation and then to the invasive character of ICC. PMID- 14563943 TI - Insights into the mechanisms of gastric adaptation to aspirin-induced injury: a role for regenerating protein but not trefoil peptides. AB - The phenomenon of reduced gastric mucosal injury despite repeated doses of a damaging agent is termed adaptation. Adaptation to nonsteroidal anti-inflammatory drug-induced injury has been clearly demonstrated in both humans and experimental animals; however, the precise mechanisms remain unclear. We hypothesized that mediators of adaptation might be the regenerating protein (RegI) and the trefoil peptides TFF1 and TFF2, because these proteins play pivotal roles in gastric mucosal protection and repair. The gene expression and the protein levels of these proteins were measured and compared in normal, aspirin-injured, and aspirin adapted rat stomachs. TFF gene and protein expression levels were similar in all three groups, whereas RegI gene expression and protein levels in adapted stomach were increased. A time course analysis of RegI expression during the onset and offset of adaptation showed that mucosal RegI increased during the development of adaptation, was maintained during subsequent aspirin dosing, and returned to baseline levels once dosing had ceased and adaptation was lost-indicative of a causal role in the adaptation process. Colocalization of increased RegI with gastric epithelial areas showing increased proliferation also suggests that RegI may be an important mediator of the resolution of mucosal injury that is characteristic of gastric adaptation to aspirin. PMID- 14563944 TI - A tissue fixative that protects macromolecules (DNA, RNA, and protein) and histomorphology in clinical samples. AB - Preservation of macromolecules (DNA, RNA, and proteins) in tissue is traditionally achieved by immediate freezing of the sample. Although isolation of PCR-able RNA has been reported from formalin-fixed, paraffin-embedded tissues, the process has not been shown to be reproducible because high molecular weight RNA is usually degraded. We investigated the potential value of a new universal molecular fixative (UMFIX, Sakura Finetek USA, Inc., Torrance, California) in preservation of macromolecules in paraffin-embedded tissue. Mouse and human tissues were fixed in UMFIX from 1 hour to 8 weeks. They were then processed by a rapid tissue processing (RTP) system, embedded in paraffin, and evaluated for routine histology as well as for the quality and quantity of DNA, RNA, and proteins. Formalin-fixed tissues were processed by RTP and evaluated in a similar manner. Fresh-frozen samples were used as controls. The morphology of UMFIX exposed tissue was comparable to that fixed in formalin. High molecular weight RNA was preserved in tissue that was immediately fixed in UMFIX and stored from 1 hour to 8 weeks at room temperature. There were no significant differences between UMFIX-exposed and frozen tissues on PCR, RT-PCR, real-time PCR, and expression microarrays. Similarly, physical and antigenic preservation of proteins in UMFIX tissue was similar to fresh state. Both RNA and proteins were substantially degraded in formalin-fixed and similarly processed specimens. We concluded that it is now possible to preserve histomorphology and intact macromolecules in the same archival paraffin-embedded tissue through the use of a novel fixative and a rapid processing system. PMID- 14563946 TI - PCR-based diagnosis of enterovirus and parvovirus B19 in paraffin-embedded heart tissue of children with suspected sudden infant death syndrome. AB - The diagnosis of viral myocarditis remains difficult and generally depends on clinical and histologic criteria. Viral cultures and serology are often unrewarding with low yields. The purpose of this study was to analyze the usefulness of PCR in the rapid diagnosis of myocarditis in children. PCR was used to analyze 120 myocardial tissue samples from 60 cases of sudden infant death syndrome (SIDS) and 56 myocardial tissue samples from 36 cases with well-known causes of sudden death (11 children younger than 1 year and 25 children 1-10 years old). The myocardial tissue samples were evaluated for the presence of enteroviruses and parvovirus B19 using PCR primers designed to consensus and unique sequences of these viral genomes. Enteroviruses could be detected in 14 cases of SIDS, whereas the detection of enteroviral nucleic acid within the control group was negative. Seven cases with myocardial infection caused by parvovirus B19 were found in the SIDS group. The detection of parvoviruses in the control group of the 11 children younger than 1 year was negative, whereas 3 positive cases of parvoviruses could be detected in the control group of children from 1 to 10 years old. In the myocardial sample of one SIDS case, both enteroviruses and parvovirus B19 could be detected. Our results emphasize the importance of modern molecular biologic methods in cases of sudden infant death even when conventional histologic examination revealed no serious findings in heart muscle tissue. PMID- 14563945 TI - Cell death in MMTV-c-myc transgenic mouse mammary tumors may not be typical apoptosis. AB - Enforced expression of c-myc has been shown to serve as an apoptotic stimulus in cultured cells. Prior studies have also demonstrated that several tissues expressing c-myc transgene display a large number of dead cells, although a morphologic or biochemical verification of apoptosis in these tissues has actually not been presented. In the present study, we examined the morphologic properties of cell death in the mammary tumors developed from MMTV-c-myc transgenic mice. We found that c-myc-expressing mammary tumor cells exhibited malformation of mitochondria, characterized by an amorphous matrix with very few cristae. The mitochondria were also frequently degenerated by lysis of the matrix and cristae. The protein level of cytochrome c was much lower in the areas of c myc-expressing tumor cells compared with the adjacent tumor foci, which was previously shown to have decreased expression of c-myc, reduced frequencies of cell death, and increased frequencies of proliferating cells. In the c-myc expressing tumor areas, there were many dying or dead cells organized in clusters, termed "dead cell islands." These cells exhibited shrinkage, DNA breakage as indicated by a positive TUNEL staining, and nuclear localization of apoptosis-inducing factor, but a lack of typical apoptotic morphology, such as nuclear condensation and formation of cell membrane blebs and apoptotic bodies. Many macrophages infiltrated into these dead cell islands, engulfing the dying or dead tumor cells. In the total tumor tissue, the protein level of caspase-3 was very low, and the poly(ADP)-ribose polymerase was present mainly as the unprocessed, inactive form. Collectively, these results suggest that programmed cell death in the c-myc transgenic mammary tumor tissue may not be typical apoptosis and may involve a caspase-independent mechanism. PMID- 14563947 TI - Pattern of recruitment of immunoregulatory antigen-presenting cells in malignant melanoma. AB - The mechanism by which the immune system of a tumor-bearing host acquires tolerance toward tumor antigens is still elusive. Antigen-presenting cells (APCs) are critical regulators of the decision between immune response and tolerance. APCs that express the tryptophan-degrading enzyme indoleamine 2,3-dioxygenase (IDO) have been found to inhibit T-cell responses both in vitro and in vivo. We hypothesized that malignant tumors exploit this mechanism by recruiting IDO expressing APCs to the tumor-draining lymph nodes. To test this hypothesis, archival tissues and records of 26 cases of lymph node dissection for invasive cutaneous melanoma were obtained. IDO immunohistochemistry was performed on 14 cutaneous tumors and 328 regional lymph nodes. Abnormal accumulations of IDO positive cells with a monocytoid or plasmacytoid morphology were identified in the perisinusoidal regions of draining lymph nodes in 45% of nodes studied. Recruitment of IDO-positive cells was seen in nodes with and without malignancy. We hypothesize that these IDO-positive APCs may contribute mechanistically to acquired tolerance to tumor antigens. Immunostaining of tumor-draining lymph nodes for abnormal accumulation of IDO-expressing cells might thus constitute an adverse prognostic factor and could contribute to the decision process and the appropriate care of patients with this deadly disease. PMID- 14563948 TI - Analysis of secondary V(D)J rearrangements in mature, peripheral T cells of ataxia-telangiectasia heterozygotes. AB - Ataxia-telangiectasia (AT) is a rare recessive disease with pleiotropic involvement of the nervous and lymphoid systems. AT heterozygotes have a population frequency of about 1%, and although not manifesting any overt clinical symptoms, they have an increased mortality, mainly because of cancer and ischemic heart disease. We and others have described a mature T lymphocyte population with an altered T cell receptor surface expression ("TCR variant") that reactivates the recombination activating genes (RAG) and is expanded in the blood of patients with AT. In view of the known role of V(D)J recombination in the onset of tumorigenic translocations, we proposed that the increased RAG activity was responsible for the predisposition of AT homozygotes to develop mature-type T leukemia/lymphoma. In the present report, we used cytofluorimetry to quantify the TCR variant population and the memory/naive T-cell compartments in the blood of AT heterozygotes compared with AT patients and controls. We assessed the expression of different recombinase genes through RT-PCR/oligotyping and cytofluorometric analysis and searched for rearrangement intermediates by ligase mediated PCR in T-cell lines from four heterozygous carriers. We found the TCR variant population was increased on average 2x in AT heterozygotes (vs 10x in homozygotes) compared with controls, and naive CD4(+) T lymphocytes were reduced on average 0.5x (vs 0.1x in homozygotes). We were able to demonstrate recombinase gene expression in all four heterozygous T-cell lines, and rearrangement intermediates, indicative of ongoing V(D)J recombination, in two. These rearrangements were compatible with V-gene replacement, a mechanism of receptor editing described for Ig and TCRalpha genes, to our knowledge not previously documented for TCRbeta. In conclusion, we found that RAG reactivation and secondary V(D)J rearrangements, potential risk factors of mature-type leukemia in AT homozygotes, also take place in AT heterozygous carriers and might place this large population fraction at an increased risk of leukemia/lymphoma. PMID- 14563950 TI - Xeroderma pigmentosum group a protein and chemotherapy resistance in human germ cell tumors. AB - The exceptional sensitivity of germ cell tumors (GCTs) of adolescents and adults to chemotherapy, in particular to cisplatin, has been attributed to low levels of xeroderma pigmentosum group A protein (XPA), a crucial component of the nucleotide excision repair DNA repair pathway. In different types of solid tumors, resistance to cisplatin has been associated with enhanced expression of XPA. To assess the role of XPA levels in clinical sensitivity and resistance of GCTs to chemotherapy, immunohistochemistry was performed on tumor samples of both unselected patients before therapy and patients with fully documented clinical course before and after therapy. In the case of high XPA levels, fluorescent in situ hybridization was applied to assess the possibility of gene amplification. XPA protein levels were investigated by Western blot analysis after repeated exposure to cisplatin in different GCT-derived cell lines. Finally, XPA levels of both sensitive and cisplatin-resistant GCT cell lines were compared with cell lines derived from other neoplasms. We found that the presence of XPA protein as assessed by immunohistochemistry differs among the various histologies of GCTs. It is found more frequently and with a more homogenous staining pattern in histologic subtypes showing a more differentiated phenotype. Overall, no differences in the presence of XPA was observed between samples of tumors refractory or sensitive to chemotherapy. No XPA gene amplification was found. Interestingly, all tumors resected in relapse after chemotherapy in the refractory group stained positive for XPA. However, XPA was not induced by repeated courses of sublethal doses of cisplatin in GCT-derived cell lines in vitro, and no correlation between XPA protein levels and sensitivity to cisplatin in three GCT-derived cell lines was observed. We therefore conclude that XPA does not play a critical role in overall treatment resistance of GCTs. PMID- 14563949 TI - Pediatric tumor cells express erythropoietin and a functional erythropoietin receptor that promotes angiogenesis and tumor cell survival. AB - Erythropoietin was traditionally considered an erythroid-restricted cytokine, but recent evidence indicates a broader role for it in nonhematopoietic tissues, specifically in neural development. Pediatric solid tumors are mostly developmental in origin, and more than 50% of the solid tumors are neural in origin. We found erythropoietin receptor and erythropoietin expression in common pediatric tumor cells: neuroblastomas, Ewing's sarcoma family of tumors, pediatric brain tumors (medulloblastoma, astrocytoma, and ependymoma), Wilms tumors, rhabdomyosarcomas, and hepatoblastomas (n = 24), and in cell lines derived from some of these tumors (n = 25). Expression of erythropoietin in tumor cell lines was hypoxia-inducible. Addition of exogenous erythropoietin to tumor cell lines expressing erythropoietin receptor increased nuclear DNA binding activity of nuclear factor kappa B and increased the expression of the antiapoptotic genes bcl-1, bcl-xL, and mcl-1. Additionally, exogenous erythropoietin increased production and secretion of angiogenic growth factors, vascular endothelial growth factor, or placenta growth factor from the tumor cell lines, which promoted endothelial cell proliferation and chemotaxis. Erythropoietin receptor expression that promotes tumor cell survival and releases angiogenic growth factors in pediatric tumors has not been previously described. Therefore, a careful evaluation of the impact of erythropoietin is warranted in vivo, in xenograft models of pediatric tumors, followed by evaluation in pediatric patients with cancer. PMID- 14563951 TI - A role for caspase-1 in serum withdrawal-induced apoptosis of endothelial cells. AB - Mouse lung endothelial cells (MLEC) and HUVEC were used under serum withdrawal (SW) conditions as a model of endothelial cell (EC) apoptosis. Apoptosis was quantified by time-lapse video microscopy. Mouse lung ECs from caspase-1(-/-) mice had significantly reduced rates of SW-induced apoptosis compared with wild type mice, specifically implicating caspase-1 in proapoptotic signaling in ECs. SW conditions induced HUVEC apoptosis with concomitant activation of caspase-1. Further studies demonstrated that the caspase-1 inhibitors z-VAD and z-YVAD significantly reduced the rate of SW-induced HUVEC apoptosis. HUVEC, when transfected with caspase-1, showed a highly significant increase in apoptosis. SW was associated with increases in reactive oxygen species production that were significantly inhibited by the antioxidant N-acetyl-L-cysteine, although rates of apoptosis and caspase-1 activation were unaffected. These results demonstrate the involvement of caspase-1 in SW-induced EC apoptosis, independently of reactive oxygen species production. PMID- 14563952 TI - High frequency of genetic aberrations in enteropathy-type T-cell lymphoma. AB - To define genetic aberrations playing a role in the development of enteropathy type T-cell lymphoma (ETL), we examined 26 such tumors using a battery of 47 microsatellite markers. The most frequent aberration (seen in 40% of informative genotypes) was amplification of genomic material in region 9q34 encompassing c abl and Notch-1 gene loci. Other frequent amplifications were detected in regions 5q33.3-34 and 7q31 (both in more than 30%). Multiple losses of heterozygosity were detected in 6p24, 7p21, 17q23-25, regions containing putative tumor suppressor genes, and in the p53 locus in 17p13.1. Analysis of the pattern of occurrence of these aberrations revealed existence of two ETL subgroups: one of them characterized by the 9q34 aberration and another smaller one showing allelic imbalances in 3q27. These two aberrations were mutually exclusive. Microsatellite instability (MSI) was detected in 69% of the examined lymphomas; the percentage of MSI-positive genotypes per tumor ranged from 2% to 12%. The spectrum of genetic alterations detected showed patterns dependent on morphology. Monomorpic ETLs displayed frequently biallelic TCR-gamma gene rearrangement (p = 0078, chi(2) test). They showed a different pattern and fewer allelic imbalances (no 3q27, 4q28, 13q14, fewer 5q21, or 5q33.3-34 aberrations) and a lower frequency of MSI than pleomorphic ETLs. PMID- 14563953 TI - Monoclonal expansion with integration of high-risk type human papillomaviruses is an initial step for cervical carcinogenesis: association of clonal status and human papillomavirus infection with clinical outcome in cervical intraepithelial neoplasia. AB - To define the natural history of cervical intraepithelial neoplasia (CIN) as related to clonal status, we evaluated 20 cases of CIN1 and 18 cases of CIN2 that had been clinically followed for 7 to 48 months at Osaka University Hospital. These included 10 cases that progressed, 15 cases that persisted, and 13 cases that regressed. We analyzed the clonal status of each case by analysis of the pattern of X-chromosomal inactivation. Human papillomavirus (HPV) infection was detected by PCR-RFLP analysis. CINs that are monoclonal or infected by high-risk HPVs are more likely to progress or persist than cases that are polyclonal or negative for high-risk HPVs (p = 0.009 for monoclonal vs polyclonal, p = 0.024 for high-risk HPV positive vs negative p = 0.024). Eighteen (90%) of 20 monoclonal, high-risk HPV-associated CINs progressed or persisted, whereas 9 (60%) of 15 polyclonal or high-risk HPV-negative CINs regressed. Therefore, the combination of clonality status and high-risk type HPV infection was significantly correlated with clinical outcome (p = 0.003). The physical status of the HPV genome was evaluated in 17 cases of HPV-16 positive CINs by real-time PCR. Of those, the HPV viral genome was present in both episomal and integrated forms in 14 CINs (84%), and 12 of these cases (86%) were monoclonal in composition. By contrast, all three CINs in which the HPV genome was present in episomal form were polyclonal. In one CIN1 that was polyclonal, HPV-16 was originally present in episomal form but after 24 months, the patient developed a monoclonal CIN3 in which the HPV-16 genome was present in mixed form. These results may imply that HPV viral integration into the host genomic DNA is associated with progression from polyclonal to monoclonal status in CIN. These events may play a fundamental role in the progression from low-grade to higher grade dysplasia of the cervical mucosa. PMID- 14563954 TI - 15-deoxy-delta-12-14-PGJ2 regulates apoptosis induction and nuclear factor-kappaB activation via a peroxisome proliferator-activated receptor-gamma-independent mechanism in hepatocellular carcinoma. AB - The peroxisome proliferator-activated receptor-gamma (PPARgamma) high-affinity ligand, 15-deoxy-Delta-12,14-PGJ(2) (15d-PGJ(2)), is toxic to malignant cells through cell cycle arrest and apoptosis induction. In this study, we investigated the effects of 15d-PGJ(2) on apoptosis induction and expression of apoptosis related proteins in hepatocellular carcinoma (HCC) cells. 15d-PGJ(2) induced apoptosis in SK-Hep1 and HepG2 cells at a 50 micro M concentration. Pretreatment with the pan-caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp (OMe) fluoromethyl ketone (2-VAD-fmk), only partially blocked apoptosis induced by 40 micro M 15d PGJ(2). This indicated that 15d-PGJ(2) induction of apoptosis was associated with a caspase-3-independent pathway. 15d-PGJ(2) also induced down-regulation of the X chromosome-linked inhibitor of apoptosis (XIAP), Bclx, and apoptotic protease activating factor-1 in SK-Hep1 cells but not in HepG2 cells. However, 15d-PGJ(2) sensitized both HCC cell lines to TNF-related apoptosis-induced ligand-induced apoptosis. In SK-Hep1 cells, cell toxicity, nuclear factor-kappaB (NF-kappaB) suppression, and XIAP down-regulation were induced by 15d-PGJ(2) treatment under conditions in which PPARgamma was down-regulated. These results suggest that the effect of 15d-PGJ(2) was through a PPARgamma-independent mechanism. Although cell toxicity was induced when PPARgamma was down-regulated in HepG2 cells, NF-kappaB suppression and XIAP down-regulation were not induced. In conclusion, 15d-PGJ(2) induces apoptosis of HCC cell lines via caspase-dependent and -independent pathways. In SK-Hep1 cells, the ability of 15d-PGJ(2) to induce cell toxicity, NF kappaB suppression, or XIAP down-regulation seemed to occur via a PPARgamma independent mechanism, but in HepG2 cells, NF-kappaB suppression by 15d-PGJ(2) was dependent on PPARgamma. PMID- 14563955 TI - Cerebral white matter lesions, retinopathy, and stroke. PMID- 14563956 TI - Antithrombotic therapy after cerebral hemorrhages. PMID- 14563957 TI - Why should mild hyperhomocysteinemia be responsible for CAD? PMID- 14563958 TI - Crossed aphasia in a dextral with right hemispheric lesion: a functional transcranial Doppler study. PMID- 14563959 TI - Angiotensin-converting enzyme inhibitors and stroke prevention: what about the influence of atrial fibrillation and antithrombotic therapy? PMID- 14563960 TI - Amphetamines for improving stroke recovery: a systematic cochrane review. PMID- 14563961 TI - Extracranial carotid artery stenosis. AB - BACKGROUND: Carotid endarterectomy is the most common surgical procedure used to treat stenosis of the extracranial precerebral carotid artery. Data from several randomized controlled trials are available to help guide its use in specific patient subgroups. Carotid angioplasty with stenting is also being performed, and clinical trials comparing this procedure with carotid endarterectomy are in progress. SUMMARY OF REPORT: For patients with symptomatic high-grade (ie, 70% to 99%) stenosis, carotid endarterectomy is associated with an overall benefit (risk ratio estimate for the combined end point of nonfatal stroke, nonfatal myocardial infarction, or death, 0.67; 95% CI, 0.54 to 0.83). The benefit is more modest for patients with less severe stenosis (ie, 50% to 69%) and may vary with specific patient characteristics. Selected patients with asymptomatic carotid stenosis may also benefit from the operation, but it needs to be performed with very low complication rates, which can be difficult to achieve in clinical practice. Several studies of angioplasty, angioplasty with stenting, and more recently angioplasty with stenting and a so-called distal protection device have also been performed. The technology involved continues to evolve rapidly, presenting a challenge for the design and conduct of clinical trials. CONCLUSIONS: Surgical intervention for extracranial carotid stenosis remains a major potential therapeutic modality for the prevention of stroke in selected patients. Endovascular approaches continue to be evaluated in ongoing trials. PMID- 14563962 TI - Subcortical white matter infarcts: comparison of superficial perforating artery and internal border-zone infarcts using diffusion-weighted magnetic resonance imaging. AB - BACKGROUND AND PURPOSE: Because of difficulty in distinguishing between superficial perforator (SP) and internal border-zone (IB) infarcts, some studies lumped SP and IB infarcts together as so-called subcortical white matter infarcts, which might complicate the classification of infarct type and its pathogenesis. Using diffusion-weighted MRI (DWI), we made comparative analyses of clinical and neuroradiological characteristics between IB and SP infarcts to clarify the difference in underlying pathogenesis. METHODS: We selected SP and IB infarcts on DWI using templates for the identification of subcortical vascular territories. Sex, age, neurological symptoms, clinical course, risk factors, concomitant cortical spotty lesions, and degree and location of arterial occlusive disease were compared between the 2 groups. RESULTS: Among the 720 consecutive patients, 54 and 29 patients met the criteria for SP and IB infarcts, respectively. The SP group had lower initial National Institutes of Health Stroke Scale scores (2.4+/-2.1 versus 4.4+/-3.1; P=0.001), favorable clinical outcomes (P=0.001), and more frequent potential cardioembolic sources (14.8% versus 0%; P=0.046) than the IB group. Cortical spotty lesions were more frequently accompanied by SP than IB infarcts (72.2% versus 24.1%; P<0.001). IB infarcts were larger (24.2+/-10.2 versus 11.4+/-8.5 mm; P<0.001), appeared in a chainlike fashion (72.4% versus 33.3%; P=0.001), and had a higher degree of stenosis or occlusion (86.2% versus 46.3%; P=0.001) compared with SP infarcts. CONCLUSIONS: Our present study suggests that embolic pathogenesis makes a greater contribution to SP infarcts than IB infarcts. This finding may explain the difference in clinical and neuroradiological characteristics between the 2 groups. PMID- 14563964 TI - Strengthening acute stroke trials through optimal use of disability end points. AB - BACKGROUND AND PURPOSE: Suboptimal choices of primary end point for acute stroke trials may have contributed to inconclusive results. The Barthel Index (BI) and Rankin Scale (RS) have been widely used and analyzed in various ways. We sought to investigate the most powerful end point for use in acute stroke trials. METHODS: Data from the Glycine Antagonist in Neuroprotection (GAIN) International Trial were used to simulate 24 000 clinical trials exploring various patterns and magnitudes of treatment effect and thus to estimate the statistical power for a range of end points based on the BI or RS. RESULTS: RS end points were more powerful than BI end points. End points dichotomized toward the favorable extreme of either scale or adjusted according to baseline prognosis ("patient-specific" end point) were among the most powerful. Combining RS and BI in a "global" end point was also successful. Improvements in statistical power indicated that using a RS end point instead of BI > or =60 could reduce the sample size by up to 84% (95% CI, 80% to 87%), 73% (95% CI, 68% to 79%) for a patient-specific BI end point, or 81% (95% CI, 76% to 85%) for a global end point. CONCLUSIONS: The RS and global end points are preferable to BI end points; the position of the cut point is also important. Better choices of end point substantially strengthen trial power for a given trial size or allow reduced sample sizes without loss of statistical power. PMID- 14563963 TI - International variations in surgical practice for spontaneous intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: Spontaneous intracerebral hemorrhage is a major cause of death and disability, yet there is no convincing evidence of the benefit of any medical treatment and the role of surgery remains controversial. The international randomized Surgical Trial in Intracerebral Hemorrhage (STICH) provided an opportunity to assess the role of surgery within the centers taking part. METHODS: Screening logs were completed to record details of all patients assessed by the department, whether they were included in the trial, the reasons if they were not included, and whether they underwent surgery. RESULTS: Logs were returned by 42 centers and cover 704 months. They include details on 1578 patients with characteristics comparable to STICH inclusion criteria. Neurosurgeons were more likely to express clinical certainty about treatment for older patients, patients with a higher Glasgow Coma Score scale, and patients in whom the hematoma was located on the right or in the basal ganglia or thalamus. Patients for whom the neurosurgeon was certain about treatment were more likely to have the hematoma removed if they were younger (62 versus 68 years of age), had a lower Glasgow Coma Scale score (10 versus 13), and had a lobar hematoma (49% versus 40%). The operation rate varied between 74% in Lithuania and 2% in Hungary. CONCLUSIONS: The difference in operation rates could not be explained by differences in patient characteristics alone. This finding demonstrates the need for further evidence to ensure that treatment for intracerebral hemorrhage is not governed by local custom. PMID- 14563965 TI - Case scenarios to assess Australian general practitioners' understanding of stroke diagnosis, management, and prevention. AB - BACKGROUND AND PURPOSE: Stroke represents the third-leading cause of death in Western society. Prompt and appropriate intervention for those with stroke or at risk of stroke is highly dependent on general practitioners' (GPs') knowledge and referral practices. METHODS: We randomly selected 490 eligible GPs from New South Wales, Australia, to complete our self-administered questionnaire. Case scenarios were used to assess GPs' knowledge of transient ischemic attack/ stroke risk factors, stroke prevention strategies, and management of asymptomatic and symptomatic patients. RESULTS: We received 296 completed questionnaires (60% response rate). Nearly all GPs (286, 96.6%) strongly agreed or agreed that stroke is a medical emergency. Most were aware that management by multidisciplinary teams improves outcomes (strongly agree or agree, 279; 94.3%). GPs endorsed the effectiveness of aspirin and warfarin in reducing stroke morbidity. GPs also were aware of the benefit of carotid endarterectomy (CEA) for symptomatic patients with >80% carotid stenosis but were less aware of the value of CEA for symptomatic patients with moderate stenosis. Vascular surgeon was the specialist of choice for referral of patients with high-grade carotid stenosis. Few GPs reported having seen the Cochrane Collaboration reviews of CEA for symptomatic (3.0%) and asymptomatic (1.7%) patients. CONCLUSIONS: GPs were well apprised of the evidence to support CEA for symptomatic patients with high-grade carotid stenosis. Our findings, however, invite more purposeful and effective education of GPs about stroke prevention, diagnosis, and management if optimal outcomes are to be realized. PMID- 14563966 TI - Intraventricular infusion of TrkB-Fc fusion protein promotes ischemia-induced neurogenesis in adult rat dentate gyrus. AB - BACKGROUND AND PURPOSE: We have previously shown that delivery of brain-derived neurotrophic factor (BDNF) through direct intrahippocampal gene transduction with a viral vector suppresses the formation of new dentate granule cells triggered by global forebrain ischemia. Here, we investigated whether inhibition of endogenous BDNF alters ischemia-induced neurogenesis in the dentate gyrus. METHODS: Rats were subjected to 30 minutes of global forebrain ischemia and then received intraventricular infusion of either the BDNF scavenger, TrkB-Fc fusion protein, or control Hu-Fc for 2 weeks. In parallel, all animals were injected intraperitoneally with the mitosis marker 5-bromo-2'-deoxyuridine-5' monophosphate (BrdU). Animals were killed at 2 or 6 weeks after the ischemic insult, and neurogenesis was then assessed immunocytochemically with epifluorescence or confocal microscopy. RESULTS: Infusion of TrkB-Fc fusion protein gave rise to elevated numbers of ischemia-generated new neurons, double labeled with BrdU and the early neuronal marker Hu or the mature neuronal marker NeuN, in the dentate subgranular zone and granule cell layer at 2 and 6 weeks after the insult. CONCLUSIONS: Our findings provide evidence that endogenous BDNF counteracts neuronal differentiation, but not cell proliferation or survival, in ischemia-induced dentate gyrus neurogenesis. PMID- 14563967 TI - Effect on cerebral vasospasm of coil embolization followed by microcatheter intrathecal urokinase infusion into the cisterna magna: a prospective randomized study. AB - BACKGROUND AND PURPOSE: Vasospasm remains the leading cause of death and permanent neurological disability in patients with aneurysmal subarachnoid hemorrhage (SAH). The objective of our prospective randomized trial of coil embolization followed by intrathecal urokinase infusion into the cisterna magna (ITUKI therapy) was to test its effectiveness in preventing or alleviating the severity of ischemic neurological deficits caused by vasospasm. METHODS: We enrolled 110 patients with ruptured intracranial aneurysms eligible for coil embolization and randomly assigned them to embolization with (n=57) or without (n=53) ITUKI therapy performed within 24 hours of aneurysmal SAH. The incidence of symptomatic vasospasms and the clinical outcomes, based on the Glasgow Outcome Scale, 6 months after SAH onset were assessed. RESULTS: There were no side effects or adverse reactions attributable to ITUKI therapy. Symptomatic vasospasm occurred in 5 patients (8.8%) with and 16 (30.2%) without ITUKI therapy; the difference was significant (P=0.012). Although the mortality rate did not differ between the groups, patients with ITUKI therapy had significantly better outcomes than those without (P=0.036). CONCLUSIONS: Our results demonstrate that ITUKI therapy significantly reduced the occurrence of symptomatic vasospasm. Although it did not completely prevent vasospasms, ITUKI therapy resulted in a lower rate of permanent neurological deficits. PMID- 14563968 TI - Abciximab reduces monocyte tissue factor in carotid angioplasty and stenting. AB - BACKGROUND AND PURPOSE: Abciximab, a nonselective glycoprotein IIb/IIIa inhibitor, was shown to reduce peri-interventional stroke rate in carotid stenting. We evaluated the effect of adjunct abciximab therapy on monocyte platelet cross talk and neurological deficit in unprotected carotid stenting and compared its efficacy with distal filter protection. METHODS: Fifty patients were randomized to either standard antithrombotic therapy (n=30) consisting of aspirin, clopidogrel, and heparin or adjunct bolus (0.25 mg/kg) and 12-hour infusion (0.125 microg x kg(-1) x min(-1)) of abciximab (n=20). A third cohort of patients was stented with filter protection (n=30). Monocyte-platelet aggregate formation and monocyte tissue factor expression were determined by whole blood flow cytometry, and F1.2 generation and soluble CD40 ligand (sCD40L) were determined by immunoassay. RESULTS: The incidence of peri-interventional ischemic episodes (23% versus 10%; P=0.2) and the number of de novo ischemic lesions detected by diffusion-weighted MRI (47% versus 30%; P=0.17) were not significantly different between standard antithrombotic therapy and adjunct abciximab but were reduced with filter protection (P=0.023). However, the number of transient ischemic attacks was lower (P=0.05) and the National Institutes of Health Stroke Score rapidly decreased in patients with adjunct abciximab. This clinical improvement was paralleled by a reduction in the postinterventional percentage of activated monocyte-platelet aggregates (CD62P+/CD14+; P=0.018) and the number of tissue factor-positive monocytes (TF+/CD14+; P=0.005). Both abciximab and filter protection suppressed F1.2 generation and significantly reduced sCD40L. CONCLUSIONS: Abciximab limits thrombus propagation and thrombus stabilization after carotid stenting by reducing monocyte-platelet cross talk and sCD40L. Although abciximab seems inferior to filter devices in peri interventional cerebral protection, it may be considered in patients who do not allow placement of protection devices. PMID- 14563969 TI - Role of monitoring in management of acute ischemic stroke patients. AB - BACKGROUND AND PURPOSE: Although several studies have demonstrated the effectiveness of specialist Stroke Unit (SU) care of stroke patients, there is still disagreement over how these units are best organized. We sought to clarify the role of continuous monitoring of physiological parameters in acute ischemic stroke. METHODS: We conducted a prospective study of 268 first-ever ischemic stroke patients admitted to our Cerebrovascular Department and allocated, according to the availability of beds, to the SU or Cerebrovascular Unit (CU). Statistical analysis compared mortality and outcome at discharge, medical and neurological complications, and length of hospitalization in the 2 care settings. RESULTS: Two hundred sixty-eight patients were enrolled. A good outcome at discharge, observed in 114 SU patients (85%) and 78 CU patients (58%) (odds ratio, 2.63; 95% CI, 1.4 to 4.8; P<0.02), was found, on multivariate analysis, to be significantly related to type of care (SU versus CU). A significantly greater proportion of SU patients showed adverse changes in monitored parameters, which required acute medical treatment (SU: 64%; CU: 19%; P<0.0001). The mean duration of these complications was significantly shorter in the SU patients (SU: 1.0 day; CU: 2.4 days; P<0.02), and the outcome in patients experiencing complications covered by the monitoring protocol was significantly better in the SU (66%) than in the CU (35%) group (P<0.0001). CONCLUSIONS: Admission of acute stroke patients to a monitoring SU may positively influence their outcome at discharge. Confirmation of our findings in larger trials will indicate the need for a revision of the minimum requirements of SUs, with the addition of monitoring as a new requirement. PMID- 14563970 TI - Topography and temporal evolution of hypoxic viable tissue identified by 18F fluoromisonidazole positron emission tomography in humans after ischemic stroke. AB - BACKGROUND AND PURPOSE: We sought to characterize the spatial and temporal evolution of human cerebral infarction. Using a novel method of quantitatively mapping the distribution of hypoxic viable tissue identified by 18F fluoromisonidazole (18F-FMISO) PET relative to the final infarct, we determined its evolution and spatial topography in human stroke. METHODS: Patients with acute middle cerebral artery territory stroke were imaged with 18F-FMISO PET (n=19; <6 hours, 4; 6 to 16 hours, 4; 16 to 24 hours, 5; 24 to 48 hours, 6). The hypoxic volume (HV) comprised voxels with significant (P<0.05; >1 mL) uptake on statistical parametric mapping compared with 15 age-matched controls. Central, peripheral, and external zones of the corresponding infarct on the anatomically coregistered delayed CT were defined according to voxel distance from the infarct center and subdivided into 24 regions by coronal, sagittal, and axial planes. Maps ("penumbragrams") displaying the percentage of HV in each region were generated for each time epoch. RESULTS: Higher HV was observed in the central region of the infarct in patients studied within 6 hours of onset (analysis of covariance [ANCOVA]; P<0.05) compared with those studied later, in whom the HV was mainly in the periphery or external to the infarct. HV was maximal in the superior, mesial, and posterior regions of the infarct (ANCOVA; P<0.05). CONCLUSIONS: These observations suggest that infarct expansion occurs at the expense of hypoxic tissue from the center to the periphery of the ischemic region in humans, similar to that seen in experimental animal models. These findings have important pathophysiological and therapeutic implications. PMID- 14563971 TI - Stroke in 85-year-olds: prevalence, incidence, risk factors, and relation to mortality and dementia. AB - BACKGROUND AND PURPOSE: Stroke and dementia are major health problems in the elderly. We examined the prevalence and incidence of stroke and their relation to dementia in a representative sample of 494 subjects 85 years of age from Gothenburg, Sweden, who were followed up to 88 years of age. METHODS: Information on stroke was obtained from an inpatient hospital linkage system, death certificates, self-reports, and key informants. Dementia was defined according to the Diagnostic and Statistical Manual of Mental Disorders, third revision. RESULTS: The prevalence of stroke at 85 years of age was 18.8% (self-reports, 10.7%; key informants, 13.2%; register data, 13.0%). The incidence of stroke between 85 and 88 years of age was 57.2/1000 person-years (men, 32.5/1000 person years; women. 66.9/1000 person-years; self-reported, 30.8/1000 person-years; key informants, 38.5/1000 person-years; register data, 45.4/1000 person-years). Female sex (risk ratio [RR], 2.1; 95% confidence interval [CI], 1.0 to 4.8) and higher systolic blood pressure (per 10 mm Hg: RR, 1.14; 95% CI, 1.02 to 1.28) were associated with higher incidence of stroke. Baseline stroke was related to increased mortality in women and higher prevalence but not incidence of dementia. There was an association between incidence strokes and incidence dementia between 85 and 88 years of age (RR, 3.8; 95% CI, 2.2 to 6.7). CONCLUSIONS: One fifth of 85-year-olds had stroke, and half of those were demented. In this age, it is important to use several sources of information to detect stroke because of the high number of demented. High blood pressure increases stroke risk also in the very old, which is important in relation to prevention. PMID- 14563972 TI - Acute Stroke Therapy by Inhibition of Neutrophils (ASTIN): an adaptive dose response study of UK-279,276 in acute ischemic stroke. AB - BACKGROUND AND PURPOSE: UK-279,276 (neutrophil inhibitory factor) reduced infarct volume in a rat middle cerebral artery occlusion reperfusion model. ASTIN (Acute Stroke Therapy by Inhibition of Neutrophils) was an adaptive phase 2 dose response-finding, proof-of-concept study to establish whether UK-279,276 improves recovery in acute ischemic stroke. The prime objective was to determine the dose that gave a clinically relevant effect in patients. METHODS: A Bayesian sequential design with real-time efficacy data capture and continuous reassessment of the dose response allowed double-blind, randomized, adaptive allocation to 1 of 15 doses (dose range, 10 to 120 mg) or placebo and early termination for efficacy or futility. The primary end point was change from baseline to day 90 on the Scandinavian Stroke Scale (DeltaSSS), adjusted for baseline SSS, aiming for a 3-point additional mean recovery above placebo. RESULTS: Nine hundred sixty-six acute stroke patients (887 ischemic, 204 cotreated with intravenous tissue plasminogen activator; mean baseline SSS score, 28; range, 10 to 40) were treated within 6 hours of symptom onset. Mean DeltaSSS was approximately +17 points of improvement on SSS for the overall evaluable population. There was no treatment effect for UK-279,276 (posterior probability of futility, 0.89). The trial was stopped early for futility. Post hoc analysis indicated a mean 1.6-point additional improvement on DeltaSSS in the tissue plasminogen activator-treated subset (credible interval=0.5, 2.6). UK-279,276 was generally well tolerated, with no increased incidence of infections. CONCLUSIONS: UK-279,276 did not improve recovery in acute ischemic stroke patients but was devoid of serious side effects. The adaptive design facilitated early termination for futility. PMID- 14563974 TI - Agriculture and the developing world. PMID- 14563975 TI - Biodefense. Researchers await government response to self-regulation plea. PMID- 14563973 TI - Enalapril prevents impaired nitric oxide synthase-dependent dilatation of cerebral arterioles in diabetic rats. AB - BACKGROUND AND PURPOSE: Our goal was to identify the effects of chronic treatment with enalapril on cerebrovascular dysfunction and endothelial nitric oxide synthase (eNOS) protein in diabetic rats. METHODS: Rats were assigned to 1 of 4 groups: nondiabetic, diabetic, nondiabetic/enalapril-treated, and diabetic/enalapril-treated groups. Rats assigned to the nondiabetic groups were injected with vehicle (sodium citrate buffer), and rats assigned to the diabetic groups were injected with streptozotocin (50 mg/kg IP). Enalapril (10 mg/kg per day) was administered in the drinking water and coincided with the injection of vehicle or streptozotocin. Two to 3 months later, we examined responses of pial arterioles to nitric oxide synthase (NOS)-dependent agonists (acetylcholine and ADP) and a NOS-independent agonist (nitroglycerin). After these functional studies, we harvested cerebral microvessels for Western blot analysis of eNOS protein. RESULTS: We found that acetylcholine- and ADP-induced dilatation of pial arterioles was impaired in diabetic compared with nondiabetic rats. In addition, while enalapril did not alter responses in nondiabetic rats, enalapril prevented diabetes-induced impairment of NOS-dependent vasodilatation. Furthermore, eNOS protein was higher in diabetic than in nondiabetic rats, and enalapril did not produce a further increase in eNOS protein in enalapril-treated diabetic rats compared with untreated diabetic rats. CONCLUSIONS: These results suggest that enalapril prevents cerebrovascular dysfunction in diabetic rats. We speculate that the protective role of enalapril may be independent of an alteration in eNOS protein in cerebral microvessels. PMID- 14563976 TI - Biomedical research. Spain offers helping hand to hospital researchers. PMID- 14563977 TI - Stem cells. 'Stemness' genes still elusive. PMID- 14563978 TI - Evolutionary biology. Outside agitators alter wasp behavior. PMID- 14563980 TI - Geosciences. A call for telling better time over the eons. PMID- 14563981 TI - Hormesis. Sipping from a poisoned chalice. PMID- 14563979 TI - Aging research: is long life in the blood? AB - Findings published this week in the Journal of the American Medical Association indicate that long-lived people have large lipoproteins--something associated with active young adults--in their blood, and that many of them carry a gene variant that influences cholesterol balance. PMID- 14563983 TI - Nuclear physics. Proton guns set their sights on taming radioactive wastes. PMID- 14563982 TI - Hormesis. A healthful dab of radiation? PMID- 14563984 TI - 2003 Nobel Prize. Physicists honored for their medical insights. PMID- 14563985 TI - 2003 Nobel Prize. Gateways into cells usher in Nobels. PMID- 14563987 TI - 2003 Nobel Prize. Clearer forecasts for the dismal science. PMID- 14563986 TI - 2003 Nobel Prize. Cool theories garner super kudos. PMID- 14563988 TI - The threat to cone snails. PMID- 14563989 TI - Coral reef decline in the Caribbean. PMID- 14563990 TI - Comment on " 'Stemness': transcriptional profiling of embryonic and adult stem cells" and "a stem cell molecular signature". PMID- 14563991 TI - Comment on " 'Stemness': transcriptional profiling of embryonic and adult stem cells" and "a stem cell molecular signature". PMID- 14563993 TI - Public health. Grand Challenges in Global Health. AB - This week an international panel announces a list of 14 Grand Challenges in Global Health, and scientists throughout the world will be invited to submit grant proposals to pursue them with funds provided by the Bill and Melinda Gates Foundation. We describe the characteristics of these challenges and the process by which they were formulated and selected after receiving over 1000 responses to a "call for ideas" from the scientific community. PMID- 14563994 TI - Medicine. Gene therapy--new challenges ahead. AB - The successful use of retroviral gene transfer to treat 10 patients with X-linked severe combined immunodeficiency (SCID-X1) has been welcomed as evidence of the therapeutic potential of gene therapy. However, as Williams and Baum suggest in their Perspective, the discovery that 2 of the 10 patients developed leukemia within 3 years of gene therapy (Hacein-Bey-Abina et al.) reinforces the need to develop even more specific gene therapy interventions. PMID- 14563995 TI - Plant sciences. Will GM rapeseed cut the mustard? AB - Genetic modification can render crops resistant to herbicides, providing them with a growth advantage so that they can outcompete weeds. However, there are concerns about the possibility that transgenes could flow from GM crops into wild plant populations. In his Perspective, Heritage discusses these important issues in terms of a new report (Wilkinson et al.) that provides a nationwide assessment in the UK of the ability of the conventional crop, oilseed rape (Brassica napus), to form hybrids with its wild relatives. PMID- 14563996 TI - Climate change. Climate in Medieval time. AB - Many papers have referred to a "Medieval Warm Period." But how well defined is climate in this period, and was it as warm as or warmer than it is today? In their Perspective, Bradley et al. review the evidence and conclude that although the High Medieval (1100 to 1200 A.D.) was warmer than subsequent centuries, it was not warmer than the late 20th century. Moreover, the warmest Medieval temperatures were not synchronous around the globe. Large changes in precipitation patterns are a particular characteristic of "High Medieval" time. The underlying mechanisms for such changes must be elucidated further to inform the ongoing debate on natural climate variability and anthropogenic climate change. PMID- 14563997 TI - Medicine. PPARs as therapeutic targets: reverse cardiology? AB - Formation of the atherosclerotic lesions that lead to myocardial infarction is determined in part by inflammatory responses such as the production of lipid laden macrophages. In his Perspective, Plutzky highlights the parts played by PPAR nuclear receptors, especially PPAR-delta (Lee et al.), in the modulation of inflammatory responses and hence atherosclerosis. PMID- 14563998 TI - Seeing more clearly: recent advances in understanding retinal circuitry. AB - Among 10 breakthroughs that Science announced at the end of 2002 was the discovery of a photosensing (melanopsin-containing) retinal ganglion cell (RGC) and its role in entraining the circadian clock. This breakthrough exemplifies the ultimate goal of neuroscience: to understand the nervous system from molecules to behavior. Light-sensing RGCs constitute one of a dozen discrete RGC populations coding various aspects of visual scenes by virtue of their unique morphology, physiology, and coverage of the retina. Interestingly, the function of the melanopsin-containing RGCs in entraining the circadian clock need not involve much retinal processing, making it the simplest form of processing in the retina. This review focuses on recent advances in our understanding of retinal circuitry, visual processing, and retinal development demonstrated by innovative experimental techniques. It also discusses the advantages of using the retina as a model system to address some of the key questions in neuroscience. PMID- 14563999 TI - Scanning human gene deserts for long-range enhancers. PMID- 14564001 TI - A hybridization model for the plasmon response of complex nanostructures. AB - We present a simple and intuitive picture, an electromagnetic analog of molecular orbital theory, that describes the plasmon response of complex nanostructures of arbitrary shape. Our model can be understood as the interaction or "hybridization" of elementary plasmons supported by nanostructures of elementary geometries. As an example, the approach is applied to the important case of a four-layer concentric nanoshell, where the hybridization of the plasmons of the inner and outer nanoshells determines the resonant frequencies of the multilayer nanostructure. PMID- 14564002 TI - Solvent-free electrolytes with aqueous solution-like conductivities. AB - Aqueous solutions are generally assumed to be superior electrolytic conductors because of the unique dielectric and fluid properties of water. We show that their conductivities can be matched by liquid electrolytes that contain no solvent. These are proton transfer salts that are liquid at ambient temperature. The high conductivities are due to the high fluidity and ionicity rather than some sort of Grotthus mechanism, although in certain cases a mobile proton population may make a non-negligible contribution. The highest conductivities have been obtained when both cations and anions contain protons. At 25 degrees C, values of >150 millisiemens per centimeter (mS x cm(-1)) appear possible; at 100 degrees C, 470 mS x cm(-1) has been measured. Because of the combination of high ionicity and proton exchange kinetics with low vapor pressure, the systems we describe also make excellent fuel cell electrolytes. PMID- 14564000 TI - LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1. AB - We have previously shown correction of X-linked severe combined immunodeficiency [SCID-X1, also known as gamma chain (gamma(c)) deficiency] in 9 out of 10 patients by retrovirus-mediated gamma(c) gene transfer into autologous CD34 bone marrow cells. However, almost 3 years after gene therapy, uncontrolled exponential clonal proliferation of mature T cells (with gammadelta+ or alphabeta+ T cell receptors) has occurred in the two youngest patients. Both patients' clones showed retrovirus vector integration in proximity to the LMO2 proto-oncogene promoter, leading to aberrant transcription and expression of LMO2. Thus, retrovirus vector insertion can trigger deregulated premalignant cell proliferation with unexpected frequency, most likely driven by retrovirus enhancer activity on the LMO2 gene promoter. PMID- 14564003 TI - Bonding changes in compressed superhard graphite. AB - Compressed under ambient temperature, graphite undergoes a transition at approximately 17 gigapascals. The near K-edge spectroscopy of carbon using synchrotron x-ray inelastic scattering reveals that half of the pi-bonds between graphite layers convert to sigma-bonds, whereas the other half remain as pi-bonds in the high-pressure form. The x-ray diffraction pattern of the high-pressure form is consistent with a distorted graphite structure in which bridging carbon atoms between graphite layers pair and form sigma-bonds, whereas the nonbridging carbon atoms remain unpaired with pi-bonds. The high-pressure form is superhard, capable of indenting cubic-diamond single crystals. PMID- 14564004 TI - Always Good Turing: asymptotically optimal probability estimation. AB - While deciphering the Enigma code, Good and Turing derived an unintuitive, yet effective, formula for estimating a probability distribution from a sample of data. We define the attenuation of a probability estimator as the largest possible ratio between the per-symbol probability assigned to an arbitrarily long sequence by any distribution, and the corresponding probability assigned by the estimator. We show that some common estimators have infinite attenuation and that the attenuation of the Good-Turing estimator is low, yet greater than 1. We then derive an estimator whose attenuation is 1; that is, asymptotically it does not underestimate the probability of any sequence. PMID- 14564005 TI - Contribution of the Patagonia Icefields of South America to sea level rise. AB - Digital elevation models of the Northern and Southern Patagonia Icefields of South America generated from the 2000 Shuttle Radar Topography Mission were compared with earlier cartography to estimate the volume change of the largest 63 glaciers. During the period 1968/1975-2000, these glaciers lost ice at a rate equivalent to a sea level rise of 0.042 +/- 0.002 millimeters per year. In the more recent years 1995-2000, average ice thinning rates have more than doubled to an equivalent sea level rise of 0.105 +/- 0.011 millimeters per year. The glaciers are thinning more quickly than can be explained by warmer air temperatures and decreased precipitation, and their contribution to sea level per unit area is larger than that of Alaska glaciers. PMID- 14564006 TI - The chemistry of sexual deception in an orchid-wasp pollination system. AB - The "sexually deceptive" orchid Chiloglottis trapeziformis attracts males of its pollinator species, the thynnine wasp Neozeleboria cryptoides, by emitting a unique volatile compound, 2-ethyl-5-propylcyclohexan-1,3-dione, which is also produced by female wasps as a male-attracting sex pheromone. PMID- 14564007 TI - Requirement of mammalian Timeless for circadian rhythmicity. AB - Despite a central circadian role in Drosophila for the transcriptional regulator Timeless (dTim), the relevance of mammalian Timeless (mTim) remains equivocal. Conditional knockdown of mTim protein expression in the rat suprachiasmatic nucleus (SCN) disrupted SCN neuronal activity rhythms, and altered levels of known core clock elements. Full-length mTim protein (mTIM-fl) exhibited a 24-hour oscillation, where as a truncated isoform (mTIM-s) was constitutively expressed. mTIM-fl associated with the mammalian clock Period proteins (mPERs) in oscillating SCN cells. These data suggest that mTim is required for rhythmicity and is a functional homolog of dTim on the negative-feedback arm of the mammalian molecular clockwork. PMID- 14564008 TI - Diffusion dynamics of glycine receptors revealed by single-quantum dot tracking. AB - Semiconductor quantum dots (QDs) are nanometer-sized fluorescent probes suitable for advanced biological imaging. We used QDs to track individual glycine receptors (GlyRs) and analyze their lateral dynamics in the neuronal membrane of living cells for periods ranging from milliseconds to minutes. We characterized multiple diffusion domains in relation to the synaptic, perisynaptic, or extrasynaptic GlyR localization. The entry of GlyRs into the synapse by diffusion was observed and further confirmed by electron microscopy imaging of QD-tagged receptors. PMID- 14564009 TI - Hematopoietic cell regulation by Rac1 and Rac2 guanosine triphosphatases. AB - The Rho guanosine triphosphatases (GTPases) Rac1 and Rac2 are critical signaling regulators in mammalian cells. The deletion of both Rac1 and Rac2 murine alleles leads to a massive egress of hematopoietic stem/progenitor cells (HSC/Ps) into the blood from the marrow, whereas Rac1-/- but not Rac2-/- HSC/Ps fail to engraft in the bone marrow of irradiated recipient mice. In contrast, Rac2, but not Rac1, regulates superoxide production and directed migration in neutrophils, and in each cell type, the two GTPases play distinct roles in actin organization, cell survival, and proliferation. Thus, Rac1 and Rac2 regulate unique aspects of hematopoietic development and function. PMID- 14564010 TI - A Bayesian networks approach for predicting protein-protein interactions from genomic data. AB - We have developed an approach using Bayesian networks to predict protein-protein interactions genome-wide in yeast. Our method naturally weights and combines into reliable predictions genomic features only weakly associated with interaction (e.g., messenger RNAcoexpression, coessentiality, and colocalization). In addition to de novo predictions, it can integrate often noisy, experimental interaction data sets. We observe that at given levels of sensitivity, our predictions are more accurate than the existing high-throughput experimental data sets. We validate our predictions with TAP (tandem affinity purification) tagging experiments. Our analysis, which gives a comprehensive view of yeast interactions, is available at genecensus.org/intint. PMID- 14564011 TI - Critical roles for Rac1 and Rac2 GTPases in B cell development and signaling. AB - The Rac1 guanosine triphosphatase (GTPase) has been implicated in multiple cellular functions, including actin dynamics, proliferation, apoptosis, adhesion, and migration resulting from signaling by multiple receptors, including the B cell antigen receptor (BCR). We used conditional gene targeting to generate mice with specific Rac1 deficiency in the B cell lineage. In the absence of both Rac1 and the highly related Rac2, B cell development was almost completely blocked. Both GTPases were required to transduce BCR signals leading to proliferation, survival and up-regulation of BAFF-R, a receptor for BAFF, a key survival molecule required for B cell development and maintenance. PMID- 14564012 TI - Listeria intracellular growth and virulence require host-derived lipoic acid. AB - Listeria monocytogenes is a Gram-positive intracytosolic pathogen that causes severe disease in pregnant and immunocompromised individuals. We found that L. monocytogenes lacking the lipoate protein ligase LplA1 was defective for growth specifically in the host cytosol and was less virulent in animals by a factor of 300. A major target for LplA1, the E2 subunit of pyruvate dehydrogenase (PDH), lacked a critical lipoyl modification when the DeltalplA1 strain was grown intracellularly, which suggests that abortive growth was due to loss of PDH function. Thus, the use of host-derived lipoic acid may be a critical process for in vivo replication of bacterial pathogens. PMID- 14564013 TI - Infectious behavior in a parasitoid. PMID- 14564014 TI - Induction of APOBEC3G ubiquitination and degradation by an HIV-1 Vif-Cul5-SCF complex. AB - Human immunodeficiency virus-1 (HIV-1) Vif is essential for viral evasion of host antiviral factor CEM15/APOBEC3G. We report that Vif interacts with cellular proteins Cul5, elongins B and C, and Rbx1 to form an Skp1-cullin-F-box (SCF)-like complex. The ability of Vif to suppress antiviral activity of APOBEC3G was specifically dependent on Cul5-SCF function, allowing Vif to interact with APOBEC3G and induce its ubiquitination and degradation. A Vif mutant that interacted with APOBEC3G but not with Cul5-SCF was functionally inactive. The Cul5-SCF was also required for Vif function in distantly related simian immunodeficiency virus mac. These results indicate that the conserved Cul5-SCF pathway used by Vif is a potential target for antiviral development. PMID- 14564016 TI - Human genome sequenced: achievements & shortcomings in big science. PMID- 14564017 TI - Particle-inflow-gun-mediated genetic transformation of buffel grass (Cenchrus ciliaris L.): optimizing biological and physical parameters. AB - The present study was conducted to optimize various biological and physical parameters for developing an efficient and reproducible gene transfer method for genetic transformation of buffel grass. Transformation was carried out using a helium-driven particle inflow gun (PIG). Embryogenic calli produced from mature seeds of buffel grass cv. CC-119 were separately bombarded with four plasmids, containing Actin (pAct1DX), Ubiquitin (pAHC-25; pAHC-27) and CaMV-35S (pCaMVGUS) promoters, coated on tungsten and gold particles. The efficiency of transformation was monitored through transient GUS expression. Different parameters, viz., the type of promoter, type and size of microcarrier, helium gas pressure, distance and time of bombardment, were standardized for delivering DNA into embryogenic calli. Bombardment with plasmid DNA carrying the actin promoter coated on 1.6 micro gold particles, at a helium pressure of 4 bars, a distance of 10 cm for 10 micro sec and 28 mm Hg vacuum in the chamber, produced the best result in transient GUS expression. The Actin promoter was found to be more efficient in driving expression of the GUS gene in buffel grass, followed by Ubiquitin and CaMV-35S promoters. Lower helium pressure was found to be sub optimal, while higher pressure produced a smaller number of blue spots, probably due to excessive damage to the cells. Maximum of 385 blue spots was observed with gold particles of 1.6 micro size, whereas only 213 blue spots were recorded for tungsten particles of 1.0 micro size. The optimized parameters can be employed for genetic transformation of buffel grass with genes of agronomic importance. PMID- 14564018 TI - Powdery mildew resistance genes in wheat: verification of STS markers. AB - Five accessions of Aegilops speltoides and 67 European wheat cultivars (winter and spring) originating from the Czech Republic, Germany, Poland, Russia, Slovakia, United Kingdom, and 4 non-European wheat cultivars from Brazil and the USA were examined with molecular Sequence Tagged Site (STS) markers for resistance genes to powdery mildew: Pm 1, Pm 2, Pm 3 and Pm 13. All markers gave clear, repeatable results, although three of them (Pm 1, Pm 2 and Pm 3) appeared as not specific for resistance genes. Comparison of STS analysis results with Pm genes, postulated as the reaction type after inoculation with differential isolates of Erysiphe graminis f.sp. tritici (Blumeria graminis), revealed a high number of disparities. The marker for Pm 13 was not detected in any examined cultivar but was present in five accessions of Aegilops speltoides. PMID- 14564015 TI - Bypassing a kinase activity with an ATP-competitive drug. AB - Unfolded proteins in the endoplasmic reticulum cause trans-autophosphorylation of the bifunctional transmembrane kinase Ire1, which induces its endoribonuclease activity. The endoribonuclease initiates nonconventional splicing of HAC1 messenger RNA to trigger the unfolded-protein response (UPR). We explored the role of Ire1's kinase domain by sensitizing it through site-directed mutagenesis to the ATP-competitive inhibitor 1NM-PP1. Paradoxically, rather than being inhibited by 1NM-PP1, drug-sensitized Ire1 mutants required 1NM-PP1 as a cofactor for activation. In the presence of 1NM-PP1, drug-sensitized Ire1 bypassed mutations that inactivate its kinase activity and induced a full UPR. Thus, rather than through phosphorylation per se, a conformational change in the kinase domain triggered by occupancy of the active site with a ligand leads to activation of all known downstream functions. PMID- 14564019 TI - RAPD-based assessment of genetic similarity and distance between Lupinus species in section Albus. AB - It was earlier suggested that the lupine taxa identified as Lupinus graecus Boissier et Spruner, L. jugoslavicus Kazim. et Now., and L. vavilovi Majss. et Atab. should be considered one species, L. albus, with two botanical varieties: var. albus (bitter cultivated form) and var. graecus (wild ancestor). We investigated this possibility by examining the genetic distance between L. albus, L. termis and L. vavilovi with RAPD markers. The genetic distances between taxa were found to be very small (<0.1), similar to results for other conspecific taxa. In addition, only three of 146 amplified fragments examined were unique to a specific taxon (L. vavilovi). We conclude that the three taxa studied belong to one species and can be divided into two varieties, as suggested earlier. PMID- 14564020 TI - Electrophoretic seed globulin patterns and species relationships in the genus Lens Miller. AB - SDS-PAGE analysis of seed globulins covered 200 accessions of the following Lens taxa: L. culinaris subsp. culinaris, L. culinaris subsp. orientalis, L. odemensis, L. ervoides, L. nigricans, L. lamottei and L. tomentosus. The number of polypeptide bands detected in particular taxa varied from 22 in L. lamottei to 35 in L. ervoides. All the taxa under study showed variation due to differences among accessions and individual variation. Electrophoretic data were subjected to statistical analysis by the UPGMA method based on Euclidean distances. In the case of L. culinaris subsp. culinaris, some distinctness of the microsperma accessions from southern and central Asia was found. As to relationships among the studied taxa, the obtained results showed that L. culinaris subsp. culinaris appeared to be most closely allied to L. odemensis, while L. culinaris subsp. orientalis was found to be the closest relative of L. tomentosus. The four taxa formed one cluster separated from L. lamottei and L. ervoides. L. nigricans was shown to be the most divergent taxon. PMID- 14564021 TI - Genetic variation of F1 hybrids from controlled crosses between Pinus montana var. rostrata and Pinus sylvestris in morphological needle traits. AB - One hundred and seventeen hybrids, representing ten families from controlled crosses between Pinus montana var. rostrata and Pinus sylvestris, were examined in respect to six morphological traits of two-year-old needles. The biometric data obtained from the measurements provided a basis for multivariate statistical analyses, including discriminant analysis and Mahalanobis distances as principal methods. The analysed families formed two complexes, which were significantly different. The main traits responsible for the distinction were numbers of stomatal rows on both sides of the needles. PMID- 14564022 TI - Non-additive genetic effects in animal selection. AB - Genetic evaluation of purebred farm animals has been carried out for about half a century, employing additive approximation to describe the genetic background. An evaluated animal has been attributed a single breeding value for each trait of the breeding goal. The predicted additive genetic value of an animal equals the average breeding value of its parents. Although the selection based on the additive approach has proved successful, there still is a possibility of increasing the reliability of the breeding value estimation by accounting for non additive genetic effects of dominance and epistasis, disregarded in the additive model. In the non-additive model, the expected quality of the progeny equals the average of the parents plus an effect resulting from the interaction between the parents. In this case, the evaluated animal may have as many breeding values as there are possible candidates to mate to, for each trait. The dominance and epistatic effects have already been accounted for in selecting animals or populations for some crossbreeding plans (combining ability, heterosis, and recombination loss). Also, using crossbreds for the sake of the breeding value estimation of purebred animals requires removing the non-additive effects from the crossbred performance and distributing the additive component between the purebreds. Combining ability is more and more discussed as a factor for matings within breed to produce terminal progeny. PMID- 14564023 TI - Sequencing of caprine alpha-S1 casein cDNAs confirms the accuracy of the RT-PCR approach for detecting of the variants of the gene. AB - Polymorphism of the casein gene in goats determines milk processing quality and cheese flavour. The main 7 alleles belong to 4 groups: strong alleles A, B, C (which code for 3.6 g/l), medium allele E (1.6 g/1), weak alleles F and D (0.6 g/1) and zero allele, connected with lack of alphaS1 casein in milk. Milk cells (mononuclear milk cells plus epithelium cells) and peripheral blood mononuclear cells were mRNA sources for reverse transcription. Three specific primers were used for polymerase chain reaction, which enabled to differentiate between alleles from four expression groups. The length of PCR products varied since allele F has a 111- nucleotide (nt) deletion of exons 9-11, allele D has a deletion of 36 nt (exon 9), and the medium allele E is associated with a 457 nt insertion in the 19th non-coding exon. Sequencing of amplified fragments, performed on PCR products isolated from milk, confirmed the correctness of the RT PCR - based alphaS1 casein genotyping method. PMID- 14564024 TI - Comparison of the nuclear organiser region activity in four taxa of the family Canidae. AB - Four species of the family Canidae were cytogenetically studied. The activity of NORs was detected with the use of silver staining. The number of NORs was characteristic for a given karyotype. For the dog found on autosomes 7, 17, 20 and on sex chromosome (Y), for the racoon dog on autosomes 1, 4, 13 and on sex chromosome (Y), for the silver fox only on autosomes 8, 9, 13 and for the blue fox on autosomes 13, 15, 17, 18, 20 and 22. The results demonstrate that NOR activity is similar in all the analysed species of the Canidae. Simultaneously, NOR activity for a medium-sized chromosome pair is distinctly higher than for two other autosome pairs (the longest and the smallest pair). Considerable variability was observed within individuals. PMID- 14564026 TI - Modern human origins and prehistoric demography of Europe in light of the present day genetic diversity. AB - Dynamic advance in DNA sequencing methods and progress in formal population genetics analyses made it possible to infer aspects of human evolution from the DNA diversity distribution and frequency in contemporary populations. While providing some general background concerning the origins of modern human, this paper focuses on the dynamics of prehistoric population in Europe. The relevance of the present-day genetic diversity studies in elucidating prehistoric events is presented in the context of archeological and paleoanthropological evidence. The questions of the Neanderthal admixture as well as of the relative contribution of different waves of prehistoric migrations to the gene pool of modern Europeans are discussed. PMID- 14564025 TI - Frequencies of alleles and genotypes of the PRNP gene in Polish Red, Czech Pied and Czech Black-and-White cattle. AB - Frequencies of alleles and genotypes of the PRNP gene in 65 Polish Red cows kept under a conservation programme and 52 randomly chosen cows of two Czech breeds: Czech Pied (42) and Black-and-White (10) were studied. All cows were at the age of 4 years or older. It was found that the frequency of allele 5 (5 copies of the octapeptide repeat) ranged from 0.11 (Polish Red) to 0.00 (Czech Black-and-White) whereas the frequency of allele 6 (6 copies of the octapeptide repeat) ranged from 1.00 (Czech Black and White) to 0.89 (Polish Red). The highest frequency was found for the homozygous genotype 6/6 (1.00 in Czech Black-and-White) and the lowest frequency was detected for the heterozygous genotype 6/5 (0.143). In the studied cows the genotype 5/5 was not found. The higher frequency of allele 5 in the native breeds and its lower frequency or lack of this allele in the populations intensively selected for high milk production may suggest that alleles of the PRNP gene may be associated with milk production. PMID- 14564027 TI - Limited pattern of TCR delta chain gene rearrangement on the RNA level in multiple sclerosis. AB - Susceptibility to multiple sclerosis (MS) is most likely affected by a number of genes, including HLA and T-cell receptor (TCR) genes. T cells expressing gamma/delta receptors seem to contribute to autoagression in MS, as evidenced by their localization in the MS plaques in the brain. The aim of this study was to analyse the TCRdelta chain gene rearrangement at the RNA (cDNA) level and compare to the DNA pattern rearrangement. TCRdelta gene rearrangement was analysed in MS patients and healthy individuals with the use of primers specific for Vdelta1-6 and Jdelta1 genes (at the DNA level) and specific for Vdelta1-6 and Cdelta1 genes (at the cDNA level). The size of PCR products was analysed on agarose gel and by ALF-Express (Pharmacia). Additionally, the lymphocyte surface immunophenotype was studied with specific monoclonal antibodies. At the DNA level a restricted pattern of Vdelta3-Jdelta1 and Vdelta5-Jdelta1 was found only in MS patients. Contrary to DNA, mono-, oligoclonal RNA (cDNA) rearrangements were limited to Vdelta1-Cdelta1, Vdelta2-Cdelta1 and Vdelta3-Cdelta1 only in MS patients as well. Surface immunophenotype analysis revealed in MS a much higher frequency of activated gamma/delta T lymphocytes, i.e. expressing HLA-DR and CD25. An elevated level of CD56 positive cells in MS was recorded. Mono-oligoclonal pattern of TCRdelta gene rearrangement at the RNA level, along with increase in activated gamma/delta T cells, strongly argue for a significant role of gamma/delta T lymphocytes in the pathogenesis of MS. PMID- 14564028 TI - The quantitative PCR technique resolves ambiguities concerning a small rearrangement of human chromosome 6q12-13. AB - Karyotype analysis, performed on the basis of chromosome banding pattern, is a standard method used for identification of chromosomal aberrations, both numerical and structural. The application of classic cytogenetic techniques fails, however, to solve all diagnostic problems in certain types of chromosome aberrations. In this study, quantitative polymerase chain reaction technique (Q PCR) application was applied to verify a cytogenetic diagnosis, which assumed that a difference observed in the banding pattern of homologous chromosome 6q12 13 region of a foetus had resulted from an inversion and/or duplication of the region in question. The obtained results indicate a possibility to use the Q-PCR method in the diagnostics of unbalanced chromosomal aberrations. PMID- 14564029 TI - Molecular follow up of donor lymphocyte infusion in CML children after allogeneic bone marrow transplantation. AB - In this study we present the applicability of fluorescence in situ hybridisation (FISH) and RT-PCR to detect the minimal residual disease (MRD) in relapsed Ph+ children after donor lymphocyte infusion (DLI) post bone marrow transplantation (BMT). In both patients BCR/ABL fusion was detected and its transcript at the moment of relapse. After the DLI treatment in short time intervals a decreasing number of cells with BCR/ABL fusion were noticed and the expression of the hybrid gene disappeared. These results demonstrate that all the methods presented in this study provide a feasible, rapid and accurate way for the detecting of the minimal residual disease after DLI in Ph positive CML patients. PMID- 14564030 TI - DNA asymmetry and the replicational mutational pressure. AB - The mode of replication and organisation of bacterial genomes impose asymmetry on their nucleotide composition. The asymmetry is seen in coding and non-coding sequences and is reflected in the amino acid composition of proteins. The mechanisms generating asymmetry include: unequal mutation rates connected with replication and transcription, selection forces positioning genes and signal sequences nonrandomly in the genome, and protein coding constraints on coding sequences. There are different methods of visualising and measuring the asymmetry. Some of them can assess the contribution of individual mechanisms to the observed asymmetry and those have been described in greater detail. Asymmetric mutational and selection pressures differentiate the rates of evolution of genes on leading and lagging strands. The genes relocated to the opposite strand have to adapt to a different mutational pressure or are eliminated. Translocations from leading to lagging strands are more often selected against than from lagging to leading strands. Comparison of intergenic sequences that have lost the coding function to the original genes enables finding the frequencies of the twelve substitution rates in sequences free from selection. In the absence of selection, the half-time of substitution of a given type of nucleotide is linearly correlated with the fraction of that nucleotide in the sequence. PMID- 14564031 TI - Meiosis and fertility of reciprocal triploid hybrids of Lolium multiflorum with Festuca pratensis. AB - Diploid and tetraploid forms of Lolium multiflorum and Festuca pratensis were crossed under controlled conditions and after embryo rescue all four combinations of autoallotriploid hybrids were obtained. Male and female fertility and chromosome pairing at metaphase I of meiosis were studied in several plants from each hybrid combination. The hybrids with two genomes of L. multiflorum and one of F. pratensis (genomic formulae LmLmFp and FpLmLm) were male and female fertile while the hybrids with two genomes of F. pratensis and one of L. multiflorum had a reduced fertility (FpFpLm) or were completely sterile (LmFpFp). Chromosome pairing at metaphase I varied among hybrid combinations depending on their genomic composition. LmLmFp and FpLmLm hybrids had similar patterns of pairing (1.83I + 5.29II + 2.85III and 2.22I + 5.22II + 2.75III, respectively) but they differed from those of FpFpLm (3.65I + 4.65II + 2.68III) and especially from LmFpFp (4.78I + 5.87II + 1.49III). Conventional analysis of meiosis failed to explain the differences in chromosome behaviour and fertility/sterility levels between the autoallotriploid hybrids with two Lolium or two Festuca genomes. PMID- 14564032 TI - Cytogenetic toxicity effects of inorganic nickel and organic Ni(II) complexes on Brassica oleracea L. root meristem. AB - Experiments were carried out on the effect of nickel as an inorganic compound (NiSO4.7H2O) and organic Ni(II) complexes (i.e. Ni(II)-Glu and Ni(II)-EDTA) in concentrations of 20, 40 and 85 ?M dm-3 on meristematic cells of root tips of Brassica oleracea L. cv. Slawa from Enkhouizen. All three tested chemical forms of nickel had a mitodepressive effect and inhibited root elongation. With respect to the degree of root elongation inhibition and mitodepressive effect, the tested forms of nickel can be put in the following order: Ni(II)-Glu NiSO4.7H2O Ni(II) EDTA. In all three tested forms, nickel caused disturbances in mitotic divisions, resulting in anaphase bridges and binuclear cells, whose nuclei were joined by a bridge of condensed chromatin or separated. Inorganic nickel and Ni(II)-Glu in higher concentrations damaged nuclei (the amount of condensed chromatin increased), nucleoli (their structure became more condensed and vacuolisation was observed), endoplasmic reticulum (fragmentation, swelling of cisternae) and mitochondria (structure condensation). PMID- 14564033 TI - Phaseolin seed variability in common bean (Phaseolus vulgaris L.) by capillary gel electrophoresis. AB - Phaseolin, the major seed storage protein of Phaseolus vulgaris from forty-four wild and cultivated accessions, was studied using sodium dodecyl sulphate capillary gel electrophoresis (SDS-CGE). In total, eleven phaseolin profiles, revealing polypeptide subunit variation in the range from 45.6 kDa to 54.4 kDa, were recorded. The number of polypeptide subunits recorded in particular profiles varied from 3 to 6; in total, eight phaseolin subunits were distinguished in the examined material. Ferguson plot analysis was used to correct non-ideal behaviour of phaseolin polypeptide subunits in capillary gel electrophoresis in the presence of SDS. The obtained results are compared to electrophoretic data received by slab polyacrylamide gel electrophoresis. The SDS-CGE method appears to provide a powerful tool for disclosure of phaseolin subunit variability. PMID- 14564034 TI - The effect of drought on the development and yielding of two different varieties of the fodder broad bean (Vicia faba minor). AB - A factorial experiment was performed in the fodder broad bean to analyse effects of soil drought on the development and yield components of two varieties of different morphotype: 'Nadwislanski' (traditional) and 'Tim' (determinate growth habit). Plants were grown in Mitscherlich's pots under three different soil moistures: 70%, 50% and 30% of field water capacity. The soil water shortage contributed to a considerable depression in the developmental characteristics and yield traits of both varieties. Under all conditions, the variety 'Nadwislanski yielded more seeds than did 'Tim'. The traditional variety was more resistant to drought than the new 'Tim'. PMID- 14564035 TI - Salt tolerance of Triticum monococcum L., T. dicoccum (Schrank) Schubl., T. durum Desf. and T. aestivum L. seedlings. AB - Responses to salt stress of diploid wheat Triticum monococcum L., tetraploid wheats T. dicoccum (Schrank) Schubl. and T. durum Desf. cv. Grandur, and hexaploid wheats T. aestivum L. cvs. Begra and Gama were studied. Seeds were germinated on filter paper in Petri dishes, moistened in Hoagland medium containing 0 (control), 10, 20, 50, 100 and 200 mM NaCl. Index of tolerance to salt treatments was calculated on the basis of shoot length. Increase in salt concentration led to decrease in seed germination and inhibition of shoot and root elongation. Hexaploid and diploid wheats were more tolerant to salt stress than tetraploid wheats. PMID- 14564036 TI - Structural and functional genomics in domestic animals: the way to understand the phenotype. AB - Molecular approaches to genome analysis in livestock are reviewed by discussing the contribution of molecular genome analysis to the identification of the genetic variation underlying phenotypic variation (structural genome analysis) and to the definition of the trait-associated and environment-affected gene expression (functional genome analysis) as an important prerequisite to understanding the formation of a phenotype. Aspects of using mapped 'quantitative trait loci' (QTL) or gene variants as well as the identified trait-associated and environment-affected gene expression profile in livestock production are expounded. PMID- 14564037 TI - Association of POU1F1/RsaI genotypes with carcass traits in pigs. AB - The objective of this study was to evaluate an association between the polymorphism of the porcine pituitary-specific transcription factor gene (POU1F1, previously called PIT1) and carcass quality in F2 animals (grandparents: Zlotnicka Spotted boars and Polish Large White sows) being a part of experimental material prepared for a QTL mapping project. The analysis covered a total of 188 F2 offspring of 13 males and 67 females (F1 generation). The RsaI PCR/RFLP polymorphism of the POU1F1 gene was identified and the least squares method was used to evaluate the significance of its effect on the value of carcass quality traits. Three POU1F1/RsaI genotypes were identified in F2 porkers: EE (n=32), EF (n=68) and FF (n=88). Twenty-four carcass quality traits were measured after 24 h of cooling. The POU1F1/RsaI genotype proved to have a significant effect on the following traits: weight of ham bone and bacon including ribs, fat thickness at the lower back (point K3), over the loin, and average fat thickness (mean of five measurements). These results confirm that the POU1F1 gene may be linked to the gene/genes affecting fat deposition in the pig carcass. Moreover, pigs with the EE genotype had a greater loin eye area and showed a higher meat weight and content of carcass than animals of both EF and FF genotypes (unsignificant association), which suggests that a further study is necessary to confirm or exclude the effect of the POU1F1 gene on these traits. PMID- 14564038 TI - Association od the DdeI growth hormone gene polymorphism with some performance traits in Polish Large White and Czech Large White x Polish Large White pigs. AB - The genotypes of growth hormone gene polymorphisms (GH-DdeI, GH-MspI, GH-HaeII, GH-ApaI, GH-CfoI) were determined in 78 pigs [Czech Large White sires (CLWsire line) x Polish Large White (PLW) sows, Polish Large White sires x Polish Large White sows], by the PCR-RFLP method. Preliminary studies found only GH DdeI polymorphism to be associated with performance traits. The associations of this polymorphism with growth and carcass traits were investigated. The linear model included the effects of candidate genes, genetic groups, sex and linear covariables of age at slaughter and body weight at weaning. The DdeI polymorphism of the GH gene showed associations with carcass length (P 100 pogeny), a higher proportion of missing records could be tolerated. The results suggest that the bivariate threshold-linear animal model is useful for routine genetic evaluation of CD with incomplete field data. PMID- 14564040 TI - Length polymorphism of PCR-amplified genomic fragments of the Pregnancy Associated Glycoprotein (PAG) gene family in the pig and some other domestic and wild mammals. AB - Porcine pregnancy-associated glycoprotein genes (pPAG) are known as a multigene family, in which five members have been cloned and sequences of their cDNAs identified. Porcine PAG1 and pPAG3 genes, belonging to the pPAG1-like subfamily, both encode enzymatically inactive precursors. In contrast, cDNAs of pPAG2, pPAG4 and pPAG6 represent the pPAG2-like gene subfamily, encoding enzymatically active precursors. The objective of this study was to investigate the polymorphism of both pPAG-like gene subfamilies in the pig in comparison to other domestic species, including cattle, sheep and goat (Artiodactyla), their wild relatives (red deer and wild pig) and horse (Perissodactyla). This is the first paper indicating the polymorphism of the pPAG gene family, examined by lengths of amplified genomic fragments (PCR). Obtained PCR products were analysed in relation to five characterised cDNAs of pPAGs (pPAG1-like and/or pPAG2-like subfamilies) and according to one recognised structural exon-intron organisation of the pPAG2 gene, among at least eight pPAG2-like genes expected in the porcine genome. The highest polymorphism frequency of both pPAG1- and pPAG2-like gene subfamilies was found in the second region, exons 5 and 6 (with intron E). The length of PCR-amplified genomic fragments was approximately: 1043, 700, 600 and 193 bp. A high polymorphism frequency was found in the 3'-terminal fragment, corresponding to exons 7-9 (with introns G and H), more frequent the pPAG2-like gene subfamily. The length of PCR-amplified genomic fragments was approximately: 733, 650 and 356 bp. In contrast, PAG polymorphism was not detected in another region, encompassing exons 2-4 (with introns B and C). The length of PCR amplified genomic fragments was approximately 279 bp in all examined genomes. In conclusion, amplification of various regions of the PAG gene family presents a relatively inexpensive PCR method of animal pre-selection with different genotypes. Such a pre-selection of animals is helpful for further gene number inquiry of the PAG gene family in each animal, then in related generations. The obtained results provide a useful background for a genetic marker preparation (by Southern analysis of the PAG family) that will presumably enable an economical early selection of young animals for effective reproduction. PMID- 14564041 TI - MELAS as an example of a mitochondrial disease. AB - MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes) is a disease mainly due to a mutation at position 3243 (A --> G) in the leucine tRNA gene in mitochondrial DNA. Symptoms of the disorder are complex and the exact pathogenesis is not understood. A review of the literature on the subject is presented. PMID- 14564042 TI - Age at diagnosis of cancer as predictor of mutation occurrence in families suspected of HNPCC. AB - Analysis of significance of age at cancer diagnosis as a factor allowing identification of a subgroup of patients with a high frequency of hMSH2 and hMLH1 mutations among families that fulfil suspected HNPCC criteria was performed. DNA from thirty-one unrelated patients affected by colorectal cancer from families matching the above criteria were studied by direct sequencing for occurrence of hMSH2 and hMLH1 gene mutations. Seven unequivocal constitutional mutations were detected: five in the hMLH1 gene and two in the hMSH2 gene. Additionally, one hMLH1 alteration of unknown significance was found. All seven mutations were found in a subgroup of 19 patients with cancer diagnosed before the age of 50 years. In a subgroup of 12 patients with cancer diagnosed at an older age only one case with hMLH1 alteration of unknown significance was detected. Our results indicate that early age at cancer diagnosis seems to be a crucial pedigree factor in discrimination of patients with hMSH2 or hMLH1 mutations among families suspected of HNPCC and matching criteria I of ICG-HNPCC. PMID- 14564043 TI - Effect of genotype on selected clinical features of Polish cystic fibrosis adults. AB - Cystic fibrosis (CF), the most common autosomal recessive disorder of Caucasians, is caused by the mutations in the gene encoding CFTR (Cystic Fibrosis Transmembrane Conductance Regulator) protein. Until now, approximately 1000 mutations of the CFTR gene have been described. The genotype-phenotype relationships in CF are still not completely understood. This study was undertaken in an attempt to characterise the distribution of CFTR mutations and their effect on selected clinical parameters in a group of Polish CF adults. A total number of 38 adult CF patients (mean age 21.6 +/- 6.8); 18 females & 20 males were enrolled in the study. The CFTR gene identification was conducted with the use of PCR & InnoLipa-CF set. The assessed clinical parameters included: age at diagnosis, age, lung function test, X-ray scored in Brasfield score, weight & height. We found that: (1) the genotypes of the studied population were unevenly distributed (65.8%- genotype deltaF508/M), (2) a high percentage of 3849+10kbC- >T was noted, (3) patients homozygous for the deltaF508 mutation were diagnosed significantly earlier and had a lower body mass index, (4) no differences were observed in the patients' length of life or the progression of lung disease. CONCLUSIONS: 1. In comparison to other populations, Polish adult CF patients display a relatively higher frequency of mild mutations. 2. Late diagnosis of CF in the studied group may be partially caused by a high percentage of CFTR mutations connected with the mild course of the disease that are difficult to identify. 3. Cystic fibrosis should be more commonly taken into consideration in the differential diagnosis in adult patients with milder symptoms. PMID- 14564044 TI - Family with Li-Fraumeni syndrome and no evidence of a germline mutation of the p53 gene or chromosomal aberrations. AB - Li-Fraumeni syndrome is a rare autosomal, dominant trait of diverse types of cancers in children and young adults, with a predominance of soft tissue sarcomas, osteosarcomas, brain tumours, adrenocortical and breast carcinomas, as well as leukaemias. We present a family with an unusual cancer history fulfilling the criteria of Li-Fraumeni syndrome. Mutational analysis of the p53 gene in constitutional DNA of several affected members of the family did not show any germline p53 defect. Cytogenetic studies did not reveal any structural aberrations. PMID- 14564045 TI - The minimal genome paradox. AB - The concept of a 'minimal genome' has appeared as an attempt to answer the question what the minimum number of genes or minimum amount of DNA to support life is. Since bacteria are cells bearing the smallest genomes, it has been generally accepted that the minimal genome must belong to a bacterial species. Currently the most popular chromosome in studies on a minimal genome belongs to Mycoplasma genitalium, a parasite bacterium whose total genetic material is as small as approximately 580 kb. However, the problem is how we define life, and thus also a minimal genome. M. genitalium is a parasite and requires substances provided by its host. Therefore, if a genome of a parasite can be considered as a minimal genome, why not to consider genomes of bacteriophages? Going further, bacterial plasmids could be considered as minimal genomes. The smallest known DNA region playing the function of the origin of replication, which is sufficient for plasmid survival in natural habitats, is as short as 32 base pairs. However, such a small DNA molecule could not form a circular form and be replicated by cellular enzymes. These facts may lead to an ostensibly paradoxical conclusion that the size of a minimal genome is restricted by the physical size of a DNA molecule able to replicate rather, than by the amount of genetic information. PMID- 14564046 TI - Molecular markers for leaf rust resistance genes in wheat. AB - Over 100 genes of resistance to rust fungi: Puccinia recondita f. sp. tritici, (47 Lr - leaf rust genes), P. striiformis (18 Yr - yellow rust genes) and P. graminis f. sp. tritici (41 Sr - stripe rust genes) have been identified in wheat (Triticum aestivum L.) and its wild relatives according to recent papers. Sixteen Lr resistance genes have been mapped using restriction fragments length polymorphism (RFLP) markers on wheat chromosomes. More than ten Lr genes can be identified in breeding materials by sequence tagged site (STS) specific markers. Gene Lrk 10, closely linked to gene Lr 10, has been cloned and its function recognized. Available markers are presented in this review. The STS, cleaved amplified polymorphic sequence (CAPS) and sequence characterized amplified regions (SCAR) markers found in the literature should be verified using Triticum spp. with different genetic background. Simple sequence repeats (SSR) markers for Lr resistance genes are now also available. PMID- 14564047 TI - Screening of cytoplasmic DNA diversity between and within Lupinus mutabilis Sweet and Lupinus albus sensu lato by restriction fragment length polymorphism (RFLP). AB - Seven populations and five mutant lines of the Andean lupin and four species from the section Albus were screened for their mitochondrial and chloroplast polymorphisms. For this purpose the RFLP method with EcoRI as a restriction enzyme was used. Lupinus luteus, Lupinus albus and Phaseolus vulgaris organellar clones as well as amplified fragments were used as probes. We found that mitochondrial probes were more suitable than chloroplast probes for identification of inter- and intra-specific variations within the examined material. Most mitochondrial probes differentiate the two species investigated. A high level of mitochondrial polymorphism was observed among the populations of L. mutabilis in contrast to monomorphism among the species in the section Albus. A limited polymorphism was detected between the mutant lines of L. mutabilis. We conclude from this study that the mitochondrial RFLP analysis is a valuable tool for identification of variability among Andean lupin populations. PMID- 14564048 TI - Inter-simple sequence repeat (ISSR) markers for the Ns resistance gene in potato (Solanum tuberosum L.). AB - Inter-simple sequence repeat (ISSR) polymorphism was used for finding markers linked to the Ns gene, responsible for a resistance of potato (Solanum tuberosum L.) to potato virus S (PVS). The ISSR markers UBC811(660) and UBC811(950) were found to be linked to Ns. Linkage distances were estimated to be 2.6 cM and 6.6 cM, respectively. UBC811(660) showed high accuracy for detection of PVS resistance in diploid potato clones. In tetraploids, among seventeen studied genotypes containing the resistance gene, this marker was revealed in eleven. UBC811(660) can be a powerful tool for detection of genotypes carrying the Ns gene in diploid potato breeding programmes. PMID- 14564049 TI - A pot evaluation of the sensitivity of spring barley (Hordeum vulgare L.) to water stress applied at different growth stages. AB - Two-year pot experiments with three varieties of spring barley were carried out. Water stress (water deficit of soil up to 40% of field water-holding capacity) was employed in four basic growth stages of plants: tillering, shooting (stem extension stage), ear formation and milk maturity. Reactions of the plants to water stress were expressed by a decline in the grain yield of the studied varieties throughout the growing season. The greatest losses in production of spring barley grain due to the experienced water stress of the plants were found at the beginning of ear formation and milk maturity stages of the grain. The sensitivity of the plants to the stress caused by water deficit in the soil during the initial period of their vegetative growth was smaller. Varieties showed significantly different sensitivities to the experienced water stress, depending on the growth stage during which the plants suffered from the stress. PMID- 14564050 TI - The possible role of cell cycle stage in mammalian cloning. AB - Progress in mammalian cloning started from cloning embryos (of mice, rats, rabbits, sheep, goats, pigs, cattle and rhesus monkeys) and culminated in obtaining clones of sheep, cattle, pigs and mice from adult somatic cells. Knowing the relationship between the cell cycles of the recipient and the donor of cell nucleus in embryonic cloning by nuclear transfer one can adjust the phases of the cell cycle properly. Metaphase II recipients accept G1 (in most species) or G2 donors (in the mouse). Interphase recipients can harbour nuclei in all stages of cell cycle. Relatively little is known about somatic cloning. Two attitudes are applied: either the donor is in the G0 phase or the recipient is in a prolonged MII phase. PMID- 14564051 TI - Application of the covariance function approach with an iterative two-stage algorithm to the estimation of parameters of a random regression test day model for dairy production traits. AB - The covariance function approach with an iterative two-stage algorithm of LIU et al. (2000) was applied to estimate parameters for the Polish Black-and-White dairy population based on a sample of 338 808 test day records for milk, fat, and protein yields. A multiple trait sire model was used to estimate covariances of lactation stages. A third-order Legendre polynomial was subsequently fitted to the estimated (co)variances to derive (co)variances of random regression coefficients for both additive genetic and permanent environment effects. Daily and 305-day heritability estimates obtained are consistent with several studies which used both fixed and random regression test day models. Genetic correlations between any two days in milk (DIM) of the same lactation as well as genetic correlations between the same DIM of two lactations were within a biologically acceptable range. It was shown that the applied estimation procedure can utilise very large data sets and give plausible estimates of (co)variance components. PMID- 14564052 TI - Generating data in models including direct and maternal dominance effects. AB - Algorithms are presented to simulate multiple generations of animal data by a model including direct additive genetic, maternal additive genetic, direct dominance, maternal dominance and permanent environmental effects. Dominance effects were computed as parental subclasses. Testing involved five single trait models that included direct contemporary group and direct additive effects, and different combinations of maternal, permanent environmental, and dominance effects. Simulated populations included 5 generations of animals and 20 contemporary groups per generation. The base population contained 200 sires and 600 dams. Variance components were estimated by Average-Information Restricted Maximum Likelihood (AIREML). No significant bias was observed. The simulation algorithms can be used in research involving dominance models, such as evaluation of mating systems exploiting special combining abilities of prospective parents. PMID- 14564053 TI - A low-level X chromosome mosaicism in mares, detected by chromosome painting. AB - Fluorescence in situ hybridization with the use of the equine X whole chromosome painting probe was carried out on chromosome spreads originating from three mares with poor reproductive performance (infertility, miscarriage or stillbirth). The numbers of analysed spreads were high (105, 300 and 480) and in all three mares a low frequency of mosaicism was identified. The mares had the following karyotypes: 64,XX/63,X/65,XXX (93.6%/5.7%/0.7%), 64,XX/63,X (98.9%/1.1%) and 64,XX/63,X (94.3%/5.7%). The incidence and importance of the low percentage X chromosome mosaicism are discussed. PMID- 14564054 TI - Forkhead genes and human disease. AB - Forkhead, or Fox-box genes, code for winged helix transcription factors that make up a multi-gene family. Two human genetic diseases have recently been associated with loss of function of one allele of different Fox-box genes: Axenfeld-Rieger anomaly of the anterior eye chamber associated with haploinsufficiency of FOXC1 and lymphedema-distichiasis associated with haploinsufficiency of FOXC2. Earlier, both genes had been studied intensively for their transcription patterns and for the phenotypes of knockouts. These studies are reviewed and related to the phenotypes found in the two human disorders. PMID- 14564055 TI - Association of arylamine N-acetyltransferase (NAT1 and NAT2) genotypes with urinary bladder cancer risk. AB - Arylamines are known bladder carcinogens deriving from tobacco smoke and environmental pollution. Arylamines are metabolised by NAT1 and NAT2 polymorphic enzymes in reactions of carcinogen activation and detoxification. We analysed genetic polymorphisms in both NAT1 and NAT2 genes in 56 bladder cancer patients and 320 healthy patients. Peripheral blood lymphocytes were collected from each subject and genotyped for NAT1 (six alleles) and NAT2 (four alleles) by PCR-RFLP. A weak association between NAT1 and NAT2 genotypes and bladder cancer risk was found when the genotypes were estimated separately (odds ratio OR 1.2, 95%CI 0.7 2.0, and OR 1.3, 95%CI 0.7-1.9, respectively). Almost all NAT1 genotypes possessing at least one "risk" *10 allele were more frequent in the bladder cancer group than in the control group. There was also an increased frequency of "risk" genotypes along with increased cigarette smoking in bladder cancer patients. The coincidence of NAT1-fast/NAT2-slow appears as a potential risk factor for urinary bladder cancer (OR 1.5, 0.8-3.0), as compared with the other genotype combinations. PMID- 14564056 TI - Influence of diethylenetriamine (DETA) and sodium nitroprusside (NaNP) on sister chromatid exchange frequency and cell kinetics in cultured human lymphocytes. AB - Diethylenetriamine (DETA) and Sodium Nitroprusside (NaNP), the exogenous NO generating compounds, were tested for their genotoxicity in human lymphocytes in vitro using the sister chromatid exchange (SCE) technique. Both compounds were found to be inactive in inducing SCE in concentrations from 0.3 to 30 pM. However both compounds displayed an inhibiting effect on cell kinetics. PMID- 14564057 TI - A familial X/Y translocation: cytogenetic and molecular study. AB - In this study we describe a 3-generation family carrying a (X;Y)(p22.3;q11.2) translocation in seven individuals of both sexes. Molecular analysis of the aberrant (X;Y)(p22.3;q11.2) chromosome was performed by FISH using X and Y specific painting probes and also PCR amplification of the Y-specific sequences. Using these approaches it was demonstrated that the translocation resulted in a deletion of both X and Y pseudoautosomal regions. Moreover, using RBG banding it was shown that in all females the X-derivative chromosome was inactive in over 90% of mitoses. From the preliminary results obtained in this study we assumed that in this particular family the observed phenotype of the patients was caused by a deletion of the cluster of pseudoaotosomal genes responsible for the stature. More proximal loci, like STS or MRX49, were probably not deleted, since neither ichtyosis nor mental retardation was observed in this family. PMID- 14564058 TI - Genetic diversity of inbred rye lines evaluated by RAPD analysis. AB - This study presents an attempt to supply breeders of hybrid rye with more genetic information on inbred lines, using molecular markers. Eighteen polymorphic loci detected by means of the RAPD (Random Amplified Polymorphic DNA) technique and mapped on 2R-7R rye chromosomes, were applied to study genetic similarities among forty inbred lines of rye. The lines were grouped in four main clusters revealed on dendrogram, which was generally consistent with the pedigree data. Mapped RAPD markers were shown to be a useful tool for phenetic studies in rye. Additionally, a system of 20 polymorphic fragments, detected by three primers, was developed for fingerprinting of rye lines. The system of RAPD markers, which was developed in this study, should be helpful in characterisation of rye genetic stocks used for breeding. PMID- 14564059 TI - Localization of the calreticulin gene mRNA in unpollinated and pollinated styles of Petunia hybrida Hort. AB - The objective of our research on Petunia hybrida is to understand the role of calreticulin in the growth of pollen tubes in the pistil. The aim of this study was the first step: finding out whether CRT gene expression takes place in unpollinated and pollinated styles. It was revealed by in situ hybridization that the transcription of the calreticulin gene takes place in the transmitting cells of unpollinated and pollinated styles and in pollen tubes growing in vivo. The mRNA transcripts of the CRT gene were localized mainly on the surface of endoplasmic reticulum (ER) membranes, both in transmitting cells and in the tip cytoplasm of pollen tubes. The results of this study show that calreticulin can be involved in pollen - pistil interaction in vivo. PMID- 14564060 TI - Karyological characterization of sugar beet gynogenetic lines cultured in vitro. AB - Flow cytometry was used to screen ploidy levels in 47 cultured in vitro sugar beet gynogenetic lines of various origin and age, obtained after plant regeneration from unfertilized ovules. When donor plants were diploid, the majority of regenerants were found to have cells with 1C, 2C and 4C relative DNA content (mainly haploid and diploid) and there were large differences in the rate of spontaneous in vitro chromosome doubling between individual homozygous lines. Six ovule-derived lines regenerated from fertile and sterile diploid donors of forty-five lines were solid diploids from the very early stages of their in vitro cultivation, and thus could not be classified as doubled haploids. In the case of tetraploid donor plants, the gynogenetic regenerants demonstrated 2x-ploidy level. The results obtained in chimeric plants with both haploid and diploid cells indicated the possibility to overcome mixoploidy by their re-cultivation through generative shoot tip culture. The flow cytometry method confirmed data obtained by conventional microscopic chromosome counting in dividing leaf cells and was found very useful for screening of a large number of regenerants and for characterizing the process of in vitro gynogenetic lines formation in sugar beet. PMID- 14564061 TI - Towards the introducing of resistance to powdery mildew from Lycopersicon hirsutum into L. esculentum. AB - Genes of resistance to Oidium lycopersicum from Lycopersicon hirsutum LA 1775 were introduced to L. esculentum. Breeding procedures were based on a one-way programme up to the F2 generation and then four different methods were adopted to obtain F4 and BC4 populations. Screening tests among those hybrid populations were performed in a greenhouse and showed segregation for resistance to powdery mildew due to different genetic backgrounds of the families derived from four breeding methods that changed the status of the gene/genes responsible for resistance to powdery mildew. F4 and BC4 populations varied in relation to morphological traits (fruit size and weight, seed and fruit productivity, number of locules). There was a significant progress in breeding in comparison to L. hirsutum regarding fruit size and weight, and the number of locules. Values of two other traits: seed and fruit productivity, that are correlated with self- and cross-compatibility, were low and similar to L. hirsutum. Therefore, another one or two backcrosses will probably improve seed and fruit productivity. PMID- 14564062 TI - Genetic diversity among cultivars of spring barley revealed by random amplified polymorphic DNA (RAPD). AB - RAPD (random amplified polymorphic DNA) polymorphism was studied in 23 malting and non-malting spring barley cultivars included in the official list of Polish cultivated varieties. Twenty-four 10-mer primers were tested in each cultivar, giving altogether 149 amplification products, 45% of which were polymorphic. The number of polymorphic bands revealed by one primer ranged from 1 to 6, with an average of 2.8. Genetic distance for all pairs of compared varieties was estimated and a dendrogram was constructed using unweighted pair group method of arithmetic means. The genetic distance between cultivars ranged from 0.11 for cvs. Apex and Bryl to 0.62 for cvs. Orthega and Madonna. Of the seven malting cultivars only two (Brenda and Stratus) formed one group at D = 0.25. The genetic distance between cvs. Brenda and Scarlett, especially recommended for brewery, was equal to 0.34. The detected polymorphism appeared to be sufficient for assessing genetic distances between cultivars, but on the basis of this polymorphism groups of malting and non-malting cultivars were not clearly distinguished. PMID- 14564063 TI - Identification of fecundity gene (FecB) carriers using microsatellite markers and its effect on sheep weight. AB - Abstract. Altogether 115 animals representing 5 genetic groups: 3 purebred Booroola Merino, Corriedale and Olkuska, and 2 Booroola crossbreds were included in the studies. In total 6 alleles from 97 bp to 119 bp in microsatellite OarAE101, and 5 alleles from 162 bp to 174 bp in BM1329 were identified. The marker of FecB gene presence seems to be an allele of 97 bp in the case of microsatellite OarAE 101 and 162 bp in the case of BM1329. Significant differences FecB carriers (Booroola-Corriedale) and non-carriers (Corriedale) in birth weight and at weaning at 100 days (males and females from twins) as well as weight gain during the first 28 days and 100 days were found. Purebred lambs showed higher values of the investigated traits. PMID- 14564064 TI - Comparison between the G-banded karyotype of the aoudad (Ammotragus lervia) and sheep (Ovis aries). AB - Karyotypes of the aoudad and sheep were compared on the basis of G-banded chromosomes at the 450 band level. The common G-banded karyotype showed the homology of all aoudad chromosomes (2n=58) with sheep chromosomes (2n=54) or sheep chromosome arms. The results of cytogenetic investigations suggest that in this case karyotype evolution has led to reduction in chromosome number as a result of centric fusions. The formation of the first metacentric chromosome occurred in the aoudad. The homology of the G-banding pattern in sheep and aoudad suggests the conservation in linear arrangement of genetic material. Thus comparative cytogenetics can be a useful tool in gene mapping. PMID- 14564065 TI - Human mitochondria in health, disease, ageing and cancer. AB - In every human cell there are hundreds of mitochondria, which are required for oxidative phosphorylation as well as many other metabolic processes. Each mitochondrion contains approximately 5 mitochondrial DNA molecules. These circular DNAs of 16.5 kb in size contain only 39 genes. Mutations in mitochondrial DNA are responsible for many diseases. Alterations in these molecules may also play a role in ageing and in tumour formation. PMID- 14564066 TI - X-linked hypophosphatemia in Polish patients. 2. Analysis of clinical features and genotype-phenotype correlation. AB - Clinical and molecular data of 59 affected persons from 36 unrelated families with XLH (36 probands and 23 members of their families) were analysed. Characteristic phenotypic features (degree of leg deformities, growth failure, tooth abnormalities, tubular reabsorption of phosphate, serum phosphate and 1,25 dihydroxyvitamin D3 concentrations, head length and hearing defect in some cases) were assessed in relation to the type and localisation of 29 different PHEX gene mutations. The severity of clinical symptoms did not strictly depend upon the type and localisation of the PHEX gene mutation. A hearing defect was correlated with mutations in the beginning fragment, while tooth abnormalities and increased head length with the mutations in the beginning and the terminal fragment of the gene. Phosphate and vitamin D3 supplementation usually slowed progressive growth retardation and leg bowing. Our results point to the probability that alternative splicing occurs in the PHEX gene, producing several active forms of the PHEX protein. Some of them might be involved in bone turnover and dentin formation, others in renal phosphate uptake and vitamin D3 metabolism. PMID- 14564067 TI - Large-scale analysis of sequence tags in Xp11.4-11.3 and evaluation of candidate genes for X-linked ocular diseases. AB - The gene-rich region of Xp11.4-Xp11.3 was characterized by increasing the physical marker density. Sequence tags (STSs) were generated by IRS- and DOP-PCR techniques, subsequent cloning, sequencing, and creation of primer pairs for single-copy sites. A total of 224 novel STSs were collected, providing an average marker density of 18 kb in the Xp11.4-Xp11.3 region which is assumed to be approximately 4 Mb in size. Sequence analysis of generated and established STSs via data base searches identified a novel gene highly homologous with the protein phosphatase 1 inhibitor 2 (IPP-2) and two pseudogenes; all of which map to the approximately 1.5 Mb proximal region of the critical region for X-linked congenital stationary night blindness type I (CSNB1) between markers DXS993 and DXS228. Using well-defined DNA panels, 69 STSs were fine-mapped to this approximately 1.5 Mb region, providing a marker coverage of one marker per 22 kb. No allelic loss was observed when the total STS content was applied to patient DNAs by PCR-mediated amplification. However, given the association of this region with a number of inherited ocular diseases, the data presented here provide valuable tools for genetic linkage and large-scale association studies. PMID- 14564068 TI - SURF1 gene mutations in Polish patients with COX-deficient Leigh syndrome. AB - One of the most frequent forms of Leigh syndrome (LS), a severe neurodegenerative, genetically heterogenous disease, is associated with cytochrome c oxidase (COX) deficiency. No mutations in any of the 13 polypeptide subunits of human COX have been detected in LS patients. Recently, SURF1, a positional candidate gene for LS has been identified on chromosome 9q34. We present the identification of SURF1 mutations in a randomly chosen group of Polish patients with a classical form of LS. Sequence analysis revealed the presence of a novel 704T-->C transition (Met235Thr), and two recurrent dinucleotide deletions (758delCA, 845delCT), as well as one novel polymorphic 573C-->G transversion (Thr191Thr). 845delCT was identified in 66% of all our patients in homozygous or heterozygous form. Our study confirms the recent observations that SURF1 is consistently involved in disorders of the mitochondrial respiratory chain in patients with typical Leigh syndrome. PMID- 14564069 TI - Genetic and environmental correlation and path coefficient analysis of fruit yield per bush and other traits in black currants (Ribes nigrum L.). AB - Twenty-five F1 hybrid families of black currants (Ribes nigrum L.) originating from diallel crosses of five cultivars were assessed. Phenotypic correlation coefficient between full-sib family means (rFS), genetic correlation coefficient between full-sib family expected values (rG) and the environmental correlation were estimated on the basis of two years of observations for 12 quantitative characteristics studied. Evident repeatability of correlations in both years was proved. A considerable consistency between the values of genetic and phenotypic correlation coefficients was also observed. Coefficients of phenotypic and environmental correlation were significant for many pairs of traits. Most of the significant environmental correlation coefficients were found for the same pairs of characteristics for which phenotypic correlation coefficients were significant. This shows that genetic and environmental factors affected similar traits. Fruit yield expressed the largest number of significant correlations with other traits. Path coefficient analysis indicated that plant and berry size were the main factors influencing directly fruit yield in both years. It was proved that the number and length of one-year-old shoots via bush size had very large indirect effects on fruit yield per plant. PMID- 14564070 TI - Genetic determination of variability of barley doubled haploids inoculated with Fusarium culmorum (W.G.Sm.) Sacc. with regard to mycotoxin accumulation and reduction in yield traits. AB - The genetic determination of variability of barley doubled haploid (DH) lines in regard of their susceptibility to Fusarium head blight caused by Fusarium culmorum was studied. The susceptibility was evaluated in 3-year field experiment on the basis of reduction in yield traits and myotoxin accumulation in infected kernels. The following traits were analysed in inoculated and control plants: kernel number and weight per ear, 1000-kernel weight, percentage of plump kernels (>2.5 mm), deoxynivalenol (DON) content and nivalenol (NIV) content of kernels. On the basis of the obtained data, heritability coefficient (ratio of genotypic to phenotypic variance) was assessed, and genetic parameters as well as the number of effective factors were estimated. Heritability coefficients calculated from two-way analysis of variance, i.e.regarding the influence of years and year x genotype interaction, appeared to be exceptionally low and ranged from 5.2% for the reduction in plump kernels to 38.2% for the reduction in 1000-kernel weight. In the case of mycotoxin accumulation about 60% of the observed variability in NIV concentrations and 30% in DON concentration resulted form genetic differences among lines. Additive effects of genes were important for all the analysed traits. Significant effects of dominance and dominance x dominance were observed for 1000-kernel weight and percentage of plump kernels. Moreover, it was found that the observed variability in yield trait reduction resulted from segregation of 5-6 effective factors, DON contents from 4 factors, while NIV content from 5 factors. PMID- 14564072 TI - Estimation of mating-compatibility of the wild service tree (Sorbus torminalis L. Crantz) by observing pollen tube germination. AB - Cross-compatibility and self-compatibility in trees of Sorbus torminalis (L.) Crantz were evaluated. Degree of mating-compatibility between three trees was determined by means of fluorescence microscope. Self-compatibility evaluated on the basis of the pollen tube germination index showed that one tree is fully self incompatible and the other two are self-compatible. Observations of pollen tube germination showed full cross-compatibility between trees under investigation. PMID- 14564071 TI - An evaluation of some morphological traits in doubled haploid lines and their F1 hybrids of head cabbage Kamienna Glowa in the vegetative phase. AB - The internal stump length, head mass and head shape of doubled haploid (DH) lines and their F1 hybrids of head cabbage Kamienna Glowa were compared. It was found that the range of variation in the investigated traits of DH lines was higher than that of their F1 hybrids. The head mass of the DH lines indicated some level of inbreeding depression, but their F1 hybrids showed a significant effect of heterosis. Genes responsible for flattened head shape were dominant over rounded shape genes. The longer internal stump trait was dominant over the shorter one. PMID- 14564073 TI - Genetic parameters of coat colour in golden fox (Vulpes vulpes L.). AB - Genetic parameters (heritability, genetic and phenotypic correlations) of chosen coat colour traits of golden fox were estimated. 1013 animals, born on the Sniaty fox farm (Poland) in 1985-1999 were evaluated. In 1993-1999 colour type was additionally assessed for 833 animals, by detailed evaluation of coat colour on the back and sides of the body, throat colour, belly colour and the amount of silver hair. The REML method was used to estimate genetic (co)variance components. Data were transformed using the probit transformation. Heritability estimates for coat traits were low (0.04 to 0.22). Values of most of the estimated genetic parameters (h2, rG, rP) were comparable to those frequently reported for other colour types of silver fox. PMID- 14564074 TI - Prediction of meat quality from biopsy in live pigs with different RYR1 genotypes. AB - To predict meat quality after slaughter, biopsy samples were taken from musculus longissimus lumborum et thoracis of live pigs at approximately 40 kg and 80 kg of weight. The obtained values from biopsies for pH1 and EC50 (electric conductivity) were compared with measurements after slaughter at a weight of approximately 110 kg. RYR1 genotypes were determined from blood samples using PCR RFLP. Mating of Nn sows with two nn boars resulted in 72 Nn and 40 nn offspring. Significant differences between the two genotypes were found for pH1 and EC50 values for the three weights. The coefficients of correlation for the Nn genotype of the RYR1 gene between the values after slaughter and both the first and the second biopsy for pH1 and EC50 were very low (r = 0.06, r = 0.14; and r = 0.26, r = 0.26; P ? 0.05). For the nn genotype were r = -0.23, r = -0.15; and r = -0.25, r = -0.11 respectively. The values of pH1 and EC50 were highly correlated (r = 0.52 to -0.84; P < or = 0.001) both within biopsies and after slaughter. PMID- 14564075 TI - Analysis of single gene multitrait effects in livestock by the use of Gibbs sampling. AB - The paper presents a method of multivariate data analysis described by a model which involves fixed effects, additive polygenic individual effects and the effects of a major gene. To find the estimates of model parameters, the maximization of likelihood function method is applied. The maximum of likelihood function is computed by the use of the Gibbs sampling approach. In this approach, following the conditional posterior distributions, values of all unknown parameters are generated. On the basis of the obtained samples the marginal posterior densities as well as the estimates of fixed effects, gene frequency, genotypic values, major gene, polygenic and error (co)variances are calculated. A numerical example, supplemented to theoretical considerations, deals with data simulated according to the considered model. PMID- 14564076 TI - Frequent D-loop polymorphism in mtDNA enables genotyping of 1400-year-old human remains from Merowingian graves. AB - Improvements of DNA extraction and amplification techniques presently enable DNA analysis of ancient DNA (aDNA) from samples which range from several hundred years of age up to possibly 5000 years. Taking advantage of the abundance of mitochondrial DNA and its polymorphic D-loop sequence, ten individuals from multiple burial sites of the Merowingian culture (South Germany), estimated to be about 1400 years old, were genotyped to determine possible kinship. Moreover, gonosomal DNA markers from the X- and Y-chromosome were applied for sex determination of the remains. In all individuals investigated, deviations from the Anderson mtDNA consensus sequence were observed, all representing substitutions (7 transitions and 3 transversions). Although such mutations have been reported from recent populations, our study constitutes the first description of these mtDNA mutations from numerous aDNA samples recovered from multiple burial sites. The results obtained by molecular anthropology can aid in describing kinship relations and burial customs of ancient remains. PMID- 14564077 TI - X-linked hypophosphatemia in Polish patients. 1. Mutations in the PHEX gene. AB - We present twenty-nine PHEX gene mutations extending our previous work, giving it to a total of 37 different mutations identified in Polish patients with familial or sporadic X-linked hypophosphatemia. Deletions, insertions and nucleotide substitutions leading to frameshift (27%), stop codon (29%), splice site (24%), and missense mutations (20%) were found. The mutations are distributed along the gene; exons 3, 4, 11, 12, 14, 15, 17, 20 and 22 are regions with the most frequent mutation events. Four mutations, P534L, G579R, R549X and IVS15+1nt, recurred in three, four, two and three unrelated patients, respectively. They have also been detected in affected persons from other countries. Twenty-eight mutations are specific for Polish population and almost all of them are unique. Most of the identified mutations are expected to result in major changes in protein structure and/or function. PMID- 14564078 TI - Adaptor-mediated amplification of minute amounts of severely fragmented ancient nucleic acids. AB - It has been repeatedly shown that high copy number mitochondrial DNA sequences can be recovered from ancient samples. A significant increase in the volume of information available to researchers will be observed when the amplification of nuclear DNA becomes commonplace and reproducible. To this end we established a modification of the Rapid Amplification of cDNA Ends (RACE) procedure normally used for the generation of cDNA ends from adaptor-ligated expressed sequence tag libraries. The modifications were designed to specifically address the problems associated with the highly damaged nucleic acids extracted from palaeontological specimens. For this study we used 6 human samples dating to 450 AD and approximately 6.500 BP that were refractory to reliable amplification of single copy loci by PCR. Racemate contents (ratio of D/L enantiomers) of aspartic acid, alanine, and leucine also indicated that no amplifiable DNA is present in 5 of the 6 samples. The proposed technique allowed us (i) to amplify four X chromosomal loci from 5 human specimens, and (ii) to correct allelic drop-out phenomena at the amelogenin locus in one individual; thus showing that the threshold of 80 x 10-3 for D/Lasp as a borderline for the presence/absence of amplifiable aDNA requires reassessment. Reliability of the proposed technique (i.e. amplification of DNA sequences endogenous to the find) was validated by the application of "ancient RACE" (aRACE) to prehistoric animal samples. PMID- 14564079 TI - Collagens, the basic proteins of the human body. AB - Collagens are structural elements of many tissues in the human body. The family of collagens can be divided into fibrillar and non-fibrillar collagens. The criterion of the classification is the structure of these proteins. Mutations in the genes encoding collagens cause a variety of human diseases that include osteogenesis imperfecta, some forms of osteoporosis, chondrodysplasias, some types of Ehlers-Danlos syndrome, arterial and intracranial aneurysms, epidermolysis bullosa and the renal disease known as Alport syndrome. The detection of mutations is important both scientifically and clinically. Defining the molecular defects underlying a disorder helps in the understanding of not only the properties of the mutated protein but also the function of the normal protein. Even though many mutations in the genes encoding collagens have been described, the pathogenic consequences of some of the mutations are not fully understood. The important rationale for mutation detection is the clinical use of molecular diagnostics in genetic counselling and differential diagnosis. PMID- 14564080 TI - Sphingosine-1-phosphate signaling in endothelial activation. AB - The signaling and functions of the Endothelial Differentiation Gene (EDG) family of G protein-coupled receptors have been extensively elucidated. All the members of EDG family were shown to be receptors for lysosphingolipids or lysophospholipids. EDG-1, the prototype of EDG family receptors, is a high affinity receptor for serum-borne bioactive lipid, Sphingosine-1-phosphate (S1P). S1P, secreted by thrombotic platelets, has been shown to regulate a variety of cellular responses, including survival, cytoskeletal remodeling, chemotaxis etc, via the activation of cell surface EDG receptors. Recently, a novel function of S1P in modulating angiogenic response has been demonstrated. This review will focus on S1P/ EDG-1 signaling in endothelial activation, in particular in the S1P mediated adherens junctions assembly and chemotaxis in endothelial cells. PMID- 14564081 TI - Induction of atherosclerosis by human chylomicron remnants: a hypothesis. AB - Epidemiologic studies have provided support for the association between delayed remnant removal and premature atherosclerosis. Triglyceride-rich particles such as chylomicrons and chylomicron remnants that carry dietary derived fats, may play a role in the early stages of developing arteriosclerosis. Currently research focuses on these lipoprotein classes seeking distinguishing factors that causes some lipoproteins to be atherogenic while others are not. Such lipoproteins could be involved in atherogenesis directly or indirectly. Direct involvement occurs by interaction of triglyceride-rich particles with the arterial wall, possibly affecting the artery wall by oxidative stress, direct endothelial toxicity by constituents such as lysophosphatidylcholine or oxysterols, induction of prothrombotic changes, stimulation of endothelial expression of cell adhesion molecules and direct interaction with circulating blood cells. Indirect involvement refers to the influence of triglyceride-rich lipoproteins on other lipoproteins on the composition of low density lipoprotein (LDL) and high density lipoprotein (HDL) particles. We propose that in individuals with delayed removal of chylomicron remnants, the prolonged exposure of areas of endothelium that have been partially activated by turbulent flow, to specific components of the remnants, results in the endothelial cells becoming further activated and able to bind monocytes. During or shortly after the transcytosis to the intima and transformation of monocytes to macrophages, the macrophages become engorged with remnant derived lipids and form the nidus of a fatty streak. PMID- 14564082 TI - Potential role of statins in inflammation and atherosclerosis. AB - The mevalonate pathway plays a crucial role in regulation of cellular cholesterol synthesis and isoprenoid groups. The entire pathway is closely regulated by feedback from an enzyme in the cascade, 3-hydroxy-3-methyl-CoA (HMG-CoA) reductase, as well as LDL receptors. Clinically, inhibition of this pathway by statins, potent inhibitors of HMG-CoA reductase, has been shown to reduce plasma levels of LDL cholesterol and several clinical trials with this group of drugs have demonstrated a remarked improvement in cardiovascular risk reduction. Interestingly, the improvement in cardiovascular end points in those trials was superior to estimations calculated from the effect on LDL cholesterol lowering. These findings support the idea of non-lipid effects of statins in atherosclerosis. Further, recent observations using in vivo and in vitro models of atherosclerosis have shed light on their potential role for manipulation of various cellular functions via inhibition of the mevalonate pathway. Among them, recently identified inhibitory effects of statins on monocyte-endothelial interaction suggest their effect on inflammation. Herein, we discuss recent progress in this area of study, with special focus on the biological function of statins. PMID- 14564083 TI - Improvement in Japanese clinical laboratory measurements of total cholesterol and HDL-cholesterol by the US Cholesterol Reference Method Laboratory Network. AB - BACKGROUND: Accurate and precise measurements of total cholesterol (TC) and HDL cholesterol (HDL-C) are necessary for effective diagnosis and treatment of lipid disorders. We studied the impact of TC certification and HDL-C evaluation in Japanese clinical laboratories to standardize their measurements. METHODS: We selected 78 laboratories participated at least twice for TC and 46 laboratories participated twice for HDL-C in the standardization protocols developed by the Cholesterol Reference Method Laboratory Network (CRMLN). We compared the initial and subsequent results using the performance guidelines established by US National Cholesterol Education Program (NCEP). RESULTS: For TC, mean percentage bias of all participants was -0.93% and -0.49% for the initial and second rounds, respectively. Mean within-sample CV was 0.72% and 0.69% for the initial and second rounds, respectively. For HDL-C, mean percentage bias of all participants was -1.86% and -0.06% for the initial and second events, respectively. Mean among run CV was 1.56% and 1.58% for the initial and second events, respectively. CONCLUSIONS: TC accuracy in the second round than the initial round tended to improvement although statistically not significant, however in the five years follow-up, mean absolute percentage bias was reduced over time. HDL-C accuracy was statistically improved in the second event than the initial event. The precision for both TC and HDL-C did not change. This study shows CRMLN protocols contribute effectively to improvement of TC and HDL-C performance. PMID- 14564084 TI - Significance of serum circulating hepatocyte growth factor in the development of carotid atherosclerosis. AB - We investigated the relation between the serum concentration of HGF and carotid atherosclerosis. Serum concentrations of HGF were measured in 128 in-patients (mean age, 74 +/- 11 years) free from cardiac, liver and renal diseases, in addition to lung diseases, in the Medical Department of Nomura Municipal Hospital between August, 2000 and June, 2001. Carotid intima-media thickness (IMT) was evaluated by ultrasonography with a 7.5 MHz linear type B-mode probe. The results showed a significantly positive correlation between serum age and HGF concentrations in patients with carotid atherosclerosis (IMT > 1.0 mm) (r = 0.391, p = 0.005). On the other hand, there was no significant correlation between age and serum HGF concentration in those without carotid atherosclerosis (IMT 25.5 nm, Normal LDL group) and small LDL (/=65 years, and men and women had similar increases, though the baseline value in women was sharply lower (p<0.001) than in men. The GWBI rose more in patients whose blood pressure was controlled by treatment (<140/90 mm Hg) than in noncontrolled patients (6.1 vs. 4.1, p<0.0001); for all patients the decreases in systolic and diastolic blood pressures produced by telmisartan correlated significantly (p<0.001 for each) with the increases in the GWBI scores. Controlling blood pressure appears to be an important element in improving subjective health perceptions of hypertensive patients. PMID- 14564134 TI - The differences between ACE inhibitor-treated and calcium channel blocker-treated hypertensive patients. AB - Large-scale outcome trials have demonstrated that blood pressure reduction with angiotensin-converting enzyme (ACE) inhibitors or calcium channel blockers (CCBs) is associated with reduced cardiovascular complications in hypertension. Comparative trials against conventional drugs and between ACE inhibitors and CCBs have failed to reveal conclusive differences in cause-specific outcomes. Studies in high-risk patients suggest that ACE inhibitors are superior to CCBs and other drugs in protection against cardiovascular events and renal disease. Very long term prospectively collected observational data from the Glasgow Blood Pressure Clinic and the UK General Practice Research Database strongly support an advantage of ACE inhibitors over CCBs for cardiovascular morbidity and mortality. Considering all the available information, it can be concluded that the use of CCBs in the routine therapy of hypertension cannot be recommended while wider use of ACE inhibitors, along with low-dose diuretics and beta blockers, appears justified. PMID- 14564133 TI - Incidence of pedal edema formation with dihydropyridine calcium channel blockers: issues and practical significance. AB - Dihydropyridine calcium channel blockers comprise a class of powerful, well tolerated, and safe antihypertensive agents that are widely used either alone or as a key component of combination therapy for hypertension. Peripheral edema, particularly of the lower limbs, is one of the most common adverse effects of dihydropyridine calcium channel blockers and may result in the need for dose reduction or drug withdrawal, both of which can adversely affect antihypertensive efficacy. Optimal use of these important drugs will involve careful dosing and sensitivity to strategies to diminish the likelihood of edema. Diuretics, either loop or thiazide, are usually not effective in alleviating pedal edema. Angiotensin-converting enzyme inhibitors or angiotensin receptor blockers in combination with a dihydropyridine calcium channel blocker may be helpful in this regard. Some calcium channel blockers may be less likely to cause pedal edema compared with others. This paper will review existing explanations of why there may be differences. A favorable tolerability profile is of particular importance for an antihypertensive medication, since hypertension is a chronic disorder necessitating long-term treatment and patient compliance. PMID- 14564135 TI - Roundtable discussion: management of hypertension in diabetics. PMID- 14564136 TI - Cardiovascular disease: a continuing threat to homeland defense. PMID- 14564137 TI - Analysis of recent papers in hypertension. Need for more aggressive blood pressure control in diabetes. PMID- 14564138 TI - Case studies in hypertension. Adrenal carcinoma. PMID- 14564139 TI - Effects of stress and behavioral interventions in hypertension. Pain and blood pressure. PMID- 14564140 TI - Common questions and answers in the management of hypertension. Residual risk. PMID- 14564141 TI - Management of acute decompensated heart failure in the emergency department. AB - Heart failure, a disease of epidemic proportions, has a tremendous clinical and financial impact on the US health care system. With more than 5 million Americans diagnosed with heart failure and 5-year mortality approaching 50%, it is the most common cause of hospitalization in patients older than 65 years and is the single most expensive diagnosis in the US health care system. Because the average US hospital loses more than 1000 dollars per heart failure admission, effective therapies that decrease length of stay, reduce hospital costs, and prevent 30-day readmissions are needed. This article reviews the relevant pathophysiology of heart failure, discusses the newest diagnostic strategies for emergency department diagnoses, evaluates recent advances and effects of early aggressive therapies, and presents a suggested algorithm for the treatment of acutely decompensated heart failure in emergency departments. PMID- 14564142 TI - Noninvasive hemodynamic monitoring in heart failure: utilization of impedance cardiography. AB - The annual health care cost incurred by heart failure patients is about $56 billion, two thirds of which is spent on management of acutely decompensated patients. Hemodynamic evaluation is helpful for optimization of treatment, monitoring clinical outcomes, and overall prognostication of heart failure patients, but historically could only be obtained via costly invasive procedures that carry substantial risks. Impedance cardiography (ICG) is a noninvasive and economical outpatient procedure that has been shown to provide reliable hemodynamic values comparable to those obtained from pulmonary artery catheterization. Recent clinical studies involving hundreds of patients have validated the accuracy and reproducibility of ICG compared with invasive techniques. The direct and derived measurements and the clinical applications of ICG in the diagnosis and treatment of heart failure are presented, as are three clinical case studies demonstrating the utility of ICG in the hemodynamic-guided management of heart failure. ICG is a viable, noninvasive technique in early- and late-stage heart failure that provides assistance in diagnostic evaluation, longitudinal prognostication, and therapeutic decisions. PMID- 14564143 TI - The most important issue: use beta blockers. PMID- 14564144 TI - Controversies in the use of beta blockers in heart failure. AB - Recent evidence from randomized controlled trials has provided compelling evidence to support the use of beta blockers in most patients with heart failure due to systolic dysfunction. There is little disagreement about the mortality benefit provided by adding beta blockers to standard therapy, which may include angiotensin-converting enzyme inhibitors, diuretics, and sometimes digoxin. A few areas are still controversial. The authors review the available literature encompassing four of those controversial areas: 1) the comparability among beta blockers; 2) the utility of beta blockers among patients with New York Heart Association class I and class IV heart failure symptoms; 3) the impact of race on the effectiveness of beta blockers; and 4) the safety and efficacy of beta blockers among patients on concomitant therapy with angiotensin-converting enzyme inhibitors, angiotensin receptor blockers, or spironolactone. PMID- 14564146 TI - Switching between beta blockers in heart failure patients: rationale and practical considerations. AB - The clinical benefit of beta blockade has been proven in a variety of pathologic settings, including hypertension, angina pectoris, acute- and post-myocardial infarction, and congestive heart failure. However, beta blockers do not all share the same clinical outcomes with respect to efficacy or safety in many of these conditions. This is especially true in HF, where differences in reverse remodeling and effects on the periphery may be important differentiating factors leading to improved efficacy. In fact, beta blockers are a heterogeneous group of agents with respect to pharmacology, receptor biology, hemodynamic effects, and tolerability. As cardiovascular disease progresses, the issue of switching from one b blocker to another is an important consideration as to how to optimize the effectiveness of adrenergic blockade. Because of the differences among beta blockers, switching should be conducted in a manner that takes into account pharmacologic differences. For example, the similarities and differences of receptor subtype blockade of the two agents and the potential effects of ancillary properties. Two protocols for switching between carvedilol, a third generation nonselective agent with vasodilation through alpha1 blockade, and a beta1-selective agent (e.g., metoprolol, atenolol) are described to simplify the process and maximize the safety and tolerability of this procedure. The optimal selection and use of adrenergic-blocking agents in the cardiovascular continuum will assist in providing improved management while minimizing safety and tolerability concerns. PMID- 14564145 TI - Beta-blocker treatment of chronic heart failure: comparison of carvedilol and metoprolol. AB - Beta blockers have been shown to prolong survival in chronic heart failure. It is currently a matter of debate whether any beta blocker is superior to the other in terms of improving symptoms, left ventricular function, or prognosis. A number of comparative studies have been performed with metoprolol, a beta1-selective second generation beta blocker, and carvedilol, a nonselective and vasodilatative third generation beta blocker. This review will focus on the different pharmacological profiles of carvedilol and metoprolol as well as on the clinical consequences derived from these differences. The results indicate that in some studies carvedilol is superior to metoprolol in improving left ventricular ejection fraction. However, because there is no conclusive evidence that carvedilol is superior to metoprolol in terms of prognosis, it is not justified to substitute metoprolol with carvedilol. Comparative data on mortality reduction are not available before termination of the Carvedilol or Metoprolol European Trial. Nevertheless, the different effects of both beta blockers on the beta-adrenergic system have an impact on tolerability and beta-adrenergic responsiveness and thus exercise tolerance in heart-failure patients. PMID- 14564147 TI - New insight into left ventricular reverse remodeling after biventricular pacing therapy for heart failure. AB - Biventricular pacing has been consistently shown to improve symptoms and exercise capacity in patients with advanced heart failure and wide QRS complexes. Recent data have shown that biventricular pacing has the propensity to reduce left ventricular volume, a condition referred to as reverse remodeling. Since the authors initial description of successful left ventricular reverse remodeling after biventricular pacing therapy, data from multicenter trials have grown and consistently support the observation. In addition to the volumetric change, there was also progressive improvement of left ventricular systolic function and clinical status. The authors subsequent data also illustrated that the benefits on reverse remodeling and left ventricular systolic function were pacing dependent and disappeared gradually when pacing was withdrawn. With the use of tissue Doppler imaging, a substantial proportion of patients were found who had mechanical delay in the free wall. During biventricular pacing, all the segments were homogenously delayed resulting in a late, albeit synchronous, systolic contraction. PMID- 14564148 TI - Pharmacotherapy in congestive heart failure: drug absorption in the management of congestive heart failure: loop diuretics. AB - Congestive heart failure is a disease state distinguished by the regular presence of both renal and hepatic abnormalities in drug handling. One such abnormality involves flaws in the process of drug absorption. In most instances, congestive heart failure-related abnormalities in drug absorption are of inconsequential significance. However, this is not the case with loop diuretics. Loop diuretic action ordinarily tracks the rate and extent of absorption if a sufficient amount of diuretic has been given to exceed the threshold for diuretic effect. In congestive heart failure, both the rate and absolute amount of loop diuretic absorbed can be reduced as a function of the heart failure state itself. In this setting, drug dissolution characteristics can assume added significance. Furosemide is the loop diuretic with the widest intra- and interpatient variability of absorption. Alternatively, the loop diuretic torsemide is rapidly and fairly completely absorbed independent of the heart failure state. This pattern of absorption establishes it as the preferred loop diuretic in the otherwise diuretic-resistant heart failure patient. However, the exact role of torsemide in the outpatient management of congestive heart failure remains to be determined. PMID- 14564149 TI - History of heart failure. PMID- 14564150 TI - Isolated tricuspid stenosis and heart failure: a focus on carcinoid heart disease. AB - A rare clinical occurrence, heart failure in the setting of tricuspid stenosis should immediately trigger a search for underlying systemic carcinoid disease. Carcinoid tumor cells can secrete a variety of vasoactive substances that result in skin erythema, excretory diarrhea, bronchospasm, and hemodynamic instability, but these manifestations are noted only in a few patients. Right heart valvular disease is common since the vasoactive noxious substances pass through the right heart unaffected and undergo metabolism in the pulmonary circulation, thereby decreasing involvement of the left-sided valves. Localization of the carcinoid tumor followed by surgically directed valvular treatment is mandatory for relief of symptoms. In nonoperative candidates, cytotoxic chemotherapy or long-term symptomatic drug treatment with somatostatin is indicated. PMID- 14564151 TI - Raine syndrome: report of a family with three affected sibs and further delineation of the syndrome. AB - We describe three affected sibs with Raine syndrome born to a consanguineous Turkish couple. Clinical findings and post-mortem assessment are presented. We have added previously unreported meso and severe telebrachyphalangy and urogenital anomalies to the clinical spectrum. Appositional new bone formation may be mistaken for fractures and callus formation--both prenatally by ultrasound and postnatally in radiographs. Further research is required to detect the underlying metabolic and molecular defects of this autosomal recessive syndrome. PMID- 14564152 TI - Ulnar/fibular ray defect and brachydactyly in a family: a possible new autosomal dominant syndrome. AB - The ulnar-mammary syndrome (MIM 181450) includes postaxial ray defects, abnormalities of growth, delayed sexual development, and mammary and apocrine gland hypoplasia. Brachydactyly type E (MIM 113300) presents with shortening of the metacarpals and phalanges in the ulnar ray in association with moderately short stature. We describe a three-generation family with variable expression of ulnar/fibular hypoplasia, brachydactyly, ulnar ray defects and short stature. The proband had ulnar hypoplasia with missing IV-Vth fingers, fibular hypoplasia on the right, bilateral club feet, growth retardation, a hypoplastic mid-face, an ASD and hemangiomas. She had normal mammary tissue and normal sweating. The mother had short stature, midfacial hypoplasia, a hypoplastic ulna and hypoplasia of the IVth metacarpal (brachydactyly) on the right without other associated malformations. The maternal grandfather had mild bilateral fibular hypoplasia and midphalangeal brachydactyly of the IV-Vth toes. His sister had mild short stature and shortening of the IVth metacarpal of the left hand. Two-point linkage analysis with microsatellite markers spanning the Ulnar-Mammary locus at 12q24.1 did not confirm linkage. The patients may have a previously undescribed syndrome. PMID- 14564153 TI - Another adult with Meier-Gorlin syndrome--insights into the natural history. AB - The Meier-Gorlin syndrome (MGS) is a rare autosomal recessive disorder, characterized by bilateral microtia, aplasia or hypoplasia of the patellae, and severe intrauterine and post-natal growth retardation. We describe the phenotype and report the medical history of a 25-year-old woman with MGS. Her phenotypic evolution was characterized by severe growth retardation with decelerated growth of the head and subsequently a relatively small head, normal intelligence, alteration of the facial features to a more proportionate appearance, improvement of joint function and incomplete breast development. Other characteristics of her phenotype in adulthood include a cheerful personality, a high forehead and accentuated naso-labial folds, relatively very small ears, hypoplastic breasts, and normal menstruation. PMID- 14564155 TI - Temtamy-like syndrome associated with translocation of 2p24 and 9q32. AB - We describe the phenotype of a 5 year old girl with features resembling Temtamy syndrome, including agenesis of the corpus callosum, ventriculomegaly, frontal bossing, peaked eyebrows, ptosis, malformed and low set ears, a depressed nasal bridge, a long philtrum, and iris and chorioretinal colobomas. Features unique to this child include profound mental retardation, bilateral sensorineural hearing loss, agenesis of the corpus callosum, patent ductus arteriosus, ventricular septal defect, unilateral renal agenesis, neurogenic bladder and hydronephrosis. High resolution chromosome analysis demonstrated a de novo, balanced translocation [46,XX,t(2;9)(p24;q32)]; and her case has some overlapping phenotypic features with cases of monosomy for 2p. This is the first documented case of Temtamy syndrome with a specific chromosomal anomaly, and will assist with the elucidation of the syndrome's underlying genetic defect. PMID- 14564154 TI - Two brothers with Burn-McKeown syndrome. AB - We report on two brothers with normal intelligence, bilateral choanal atresia, and a characteristic pattern of facial dysmorphic features consisting of hypertelorism, lower lid coloboma, narrow palpebral fissures, prominent nasal bridge, small mouth with thin lips, and protruding ears. These features show striking similarity to patients with Burn-McKeown syndrome [Burn et al., 1992 Clin. Dysmorphol 1: 137-144] and confirm the existence of this rare condition. These brothers show some additional features that were not previously reported in patients with this syndrome including median cleft palate with oronasal fistula, preauricular tag, hypomimic face, and hypoplastic unilateral kidney, thus indicating that the clinical spectrum of this entity is broader. PMID- 14564156 TI - A case of partial trisomy 4p syndrome presenting as severe hydronephrosis in utero. AB - We describe the phenotypic features in a newborn infant with an unbalanced translocation 46,XY, der(22) inv(4) (p14p16.1) t(4;22) (p15.1;q13.31)pat. The phenotype was consistent with partial trisomy 4p syndrome. Severe bilateral hydronephrosis was diagnosed at a 31 week prenatal ultrasound scan. Both the patient phenotype and the partial trisomy are unusual, the latter due to the complex nature of the chromosomal rearrangement. PMID- 14564157 TI - Interstitial deletion of chromosome 2p16.2p21. AB - We report on a female who presents with an atrial septal defect (ASD), mild hypotelorism, a prominent nasal bridge, a long smooth philtrum, mild developmental delay and a de novo interstitial deletion of the short arm of chromosome 2p, del (2)(p16.2p21). This is the first report of a deletion in chromosome 2 involving those particular breakpoints. We propose that this may represent a new recognizable chromosomal phenotype. PMID- 14564158 TI - Teebi hypertelorism syndrome. AB - Teebi (1987) described an autosomal dominant syndrome with resemblance to craniofrontonasal dysplasia. Here we present a 2 year 5 month old girl with Teebi syndrome showing a prominent forehead, hypertelorism, mild exophthalmos, downslanting palpebral fissures, a depressed nasal bridge, a broad nasal tip, a long philtrum, natal teeth, a thin upper lip, an everted lower lip, a small chin, low-set ears, preauricular fistulas, a short neck, mild pectus excavatum, an umbilical hernia, clinodactyly of the 5th fingers with mild radial deviation of the distal phalanges of the middle fingers, mild pes adductus, an ectopic kidney, and normal psychomotor development. Her mother and her grandmother had similar features. PMID- 14564159 TI - Siblings with glaucoma, mental retardation and short stature. AB - We report a sibling pair, whose parents are distantly related, with congenital glaucoma and mental retardation. There are similarities to ter Haar syndrome, but severe mental retardation has not been described previously in that condition. PMID- 14564160 TI - Large interstitial deletion of chromosome 13q and severe short stature: clinical report and review of the literature. AB - We report a 16 year old African American female with an interstitial deletion of chromosome 13 comprising approximately 40% of the long arm of this chromosome [karyotype 46,XX, del(13)(q14.12q31.2)]. We believe that this case is interesting because of the large size of the chromosome deletion, the severe growth retardation seen in the proband and her prolonged survival. PMID- 14564161 TI - Klippel-Feil anomaly in Fanconi anemia. AB - A boy is described with Fanconi anemia (FA) and Klippel-Feil anomaly. This suggests the diagnosis of FA should be considered in patients with vertebal malformations as well as other suggestive congenital anomalies. PMID- 14564162 TI - A large deletion (1.5 Mb) encompassing the neurofibromatosis type 1 (NF1) gene in a patient with sporadic NF1 associated with dysmorphism, mental retardation, and unusual ocular and skeletal features. AB - A 20 year old male patient with sporadic neurofibromatosis type 1 (NF1) is described with a large deletion (1.5 Mb) involving the NF1 gene, dysmorphism, mental retardation, and unusual ocular and skeletal features. Several NF1 patients with a large NF1 deletion and associated dysmorphism, and a large number of neurofibromas for their age have been described. This study indicates that such large deletions can also involve flanking loci which affect ocular and skeletal development. PMID- 14564163 TI - A patient with extensive blue rubber bleb nevi associated with multiple congenital anomalies: an overlap or a new syndrome? AB - We report a case of a 13 year old boy with non-familial, multiple blue rubber bleb nevi (BRBN). He also had additional dysmorphic anomalies including a low hairline, low set ears, narrow nares, ptosis, hypotelorism, a high arched palate and downward slanting of the outer canthi. Skeletal deformities included a short, slightly webbed neck, kyphosis, cubitus valgus, non-traumatic luxation of the head of the right radius and a short left little and index fingers. He had no evidence of gastrointestinal lesions or bleeding, congenital heart or renal disease or enchondromata. The boy was mentally slow compared to his peers. He had normal chromosomes. The absence of gastrointestinal bleeding or iron deficiency anaemia is inconsistent with Bean's syndrome and the lack of enchondromata in this patient militates against the diagnosis of classical Maffucci syndrome. Some of his physical features are obviously similar to those observed in a Noonan's syndrome yet the presence of extensive BRBN would be an extremely unusual finding. Since the overall picture does not fit into any of the classical categories mentioned above, the constellation of his features has yet to find a specific syndrome. It may, however, represent an overlap between them or, possibly, a new syndrome. PMID- 14564164 TI - Fusion of vertebrae, diaphragmatic hernia and unusual facies in a girl: a possible further case of Mathieu syndrome. AB - Mathieu et al., (1993) reported a family in which a male adult and his son were both affected by unusual facies, cleft palate, congenital brevicollis, diaphragmatic hernia, gastro-oesophageal reflux, and cervical vertebral fusion. Since then, to our knowledge, no other similar cases have been reported. Here we describe a girl presenting with facial features similar to the Mathieu's patients. The association of cervical vertebral fusion, diaphragmatic hernia, and severe gastro-oesophageal reflux seems to confirm this syndrome. PMID- 14564165 TI - Craniosynostosis with tracheal sleeve: a patient with Pfeiffer syndrome, tracheal sleeve and additional malformations in whom an FGFR2 mutation was found. AB - We discuss a patient with Pfeiffer syndrome who had a tracheal sleeve and an FGFR2 mutation. In the light of our findings, and previous reports of patients with craniosynostosis that also reported similar mutations, we suggest that genomic screening for FGFR2 may be useful in cases with negative FGFR2 mutation testing. PMID- 14564166 TI - Osteofibrous dysplasia in a Japanese boy with Costello syndrome. AB - We present the case of a boy with Costello syndrome who developed osteofibrous dysplasia during a phase of growth hormone therapy. The lesion and the accompanying pain disappeared after discontinuation of the therapy. Growth hormone is a known mitogen for some neoplasms and osteofibrous dysplasia has been reported to become aggressive. Thus, although osteofibrous dysplasia in Costello syndrome has not been reported before, growth hormone therapy should be used under close supervision in children with this syndrome. PMID- 14564167 TI - Absence of specific facial dysmorphy in glycogen storage disease type III. PMID- 14564169 TI - Risk and prevention of transfusion-transmitted babesiosis and other tick-borne diseases. AB - PURPOSE OF REVIEW: Tick-borne diseases have increasingly been recognized in the United States as public health problems. The importance of tick-borne diseases has been accelerated by increases in animal populations, as well as increased human recreation in wooded environments that are conducive to tick bites. Babesiosis, usually caused by the intraerythrocytic parasite, Babesia microti and transmitted by the same tick as Lyme disease, has important transfusion implications. Although Lyme disease has not been reported from blood transfusion, newly identified tick-borne diseases such as ehrlichiosis raise additional questions about the role of the tick in transfusion-transmitted diseases. RECENT FINDINGS: The risk of transfusion-transmitted babesiosis is higher than usually appreciated and in endemic areas represents a major threat to the blood supply. Furthermore, the geographic range of B. microti is expanding, other Babesia spp. have been implicated in transfusion transmission in the western United States, and the movement of blood donors and donated blood components may result in the appearance of transfusion babesiosis in areas less familiar with these parasites. Consequently, a higher degree of clinical suspicion will allow early recognition and treatment of this important transfusion complication. SUMMARY: In endemic areas transfusion-transmitted babesiosis is more prevalent than usually believed. The extension of the geographic range of various Babesia spp. and the movement of donors and blood products around the United States has resulted in the risk extending to non-endemic areas. Clinicians should maintain a high degree of clinical suspicion for transfusion-transmitted babesiosis. PMID- 14564170 TI - Risks of transfusion-transmitted infections: 2003. AB - PURPOSE OF REVIEW: While the risks of transfusion-transmitted human immunodeficiency virus, hepatitis C virus, and human T-cell leukemia virus I/II continue to decrease, additional threats to transfusion safety are posed by emerging "new" infectious diseases. RECENT FINDINGS: Following the introduction of nucleic acid testing for human immunodeficiency virus and hepatitis C virus, the American Red Cross estimates the risk of transfusion-transmitted human immunodeficiency virus to be 1:1,215,000 (per unit transfused) and 1:1,935,000 for transfusion-transmitted hepatitis C virus. Hepatitis B virus nucleic acid testing has not been implemented, and the risk of transfusion-transmitted hepatitis B virus in the United States remains relatively high at an estimated 1:205,000. The risk of transfusion-transmitted human T-cell leukemia virus I/II is 1:2,993,000, based on Red Cross estimates. Nucleic acid testing for West Nile virus began in the United States in 2003 under an investigational new drug program. No approved laboratory tests are available to screen the blood for Chagas disease, malaria, severe acute respiratory syndrome, or variant Creutzfeldt-Jakob disease. SUMMARY: Prevention of these potential transfusion transmitted infections is addressed by deferring potential donors whose personal behaviors or travel histories place them at risk. PMID- 14564171 TI - Anaphylactic transfusion reactions. AB - PURPOSE OF REVIEW: Although anaphylactic reactions to blood products are rare, the incidence of allergic reactions to blood products is similar to the allergic reaction incidence to penicillin antibiotics, and therefore worthy of proportionate attention. Comprehensive reviews and guidelines of the management of anaphylaxis currently do not include much information on blood products. Current guidelines for the specific management of anaphylactic transfusion reactions are contradictory as to the utility of anti-IgA testing and incomplete by not offering suggestions for the management of non-IgA related reactions. RECENT FINDINGS: Anti-IgA is not responsible for most reactions. Anti-haptoglobin antibodies are responsible for more reactions than anti-IgA in Japan, but the cause of most reactions is still not known. The incidence of reactions to platelets is the highest compared with fresh frozen plasma and red blood cells. Pre-storage white blood cell reduction of platelets does not decrease the incidence of reactions, indicating that white blood cell-derived cytokines are not responsible for most reactions. SUMMARY: The increased incidence of reactions to platelets compared with fresh frozen plasma suggests that a platelet-related factor may be responsible for many of the reactions. The possible role of platelet microparticles or activated platelet membranes, which carry a negative charge similar to ionic radiocontrast media, the major cause of iatrogenic anaphylactic reactions in the hospital, is explored. PMID- 14564172 TI - New directions in plasma exchange. AB - PURPOSE OF REVIEW: This review examines the literature published on therapeutic plasma exchange during 2002. The review was performed by searching Medline for pertinent articles. RECENT FINDINGS: One hundred thirty articles were identified, of which 11 are reviewed. During the period, reviews of the use of therapeutic plasma exchange for managing Guillain-Barre syndrome and myasthenia gravis were published. A large randomized trial of the use of plasma exchange to treat sepsis also appeared. Finally, a large case series of the use of plasma exchange in Wegener granulomatosis was published. SUMMARY: The literature confirms the use of plasma exchange for Guillain-Barre syndrome but suggests that inadequate evidence exists to support its use for long-term improvement in myasthenia gravis. The study of patients with severe sepsis suggests that plasma exchange may benefit a subset of patients, those with abdominal infections. Finally, plasma exchange for Wegener granulomatosis with severe renal dysfunction appears not to offer any benefit over immunosuppressive therapy. PMID- 14564173 TI - Red blood cell transfusion in autoimmune hemolytic anemia. AB - PURPOSE OF REVIEW: Transfusion, in the setting of autoimmune hemolytic anemia, can be a complicated and potentially dangerous proposition. RECENT FINDINGS: The selection and delivery of an appropriate red blood cell unit must focus on several areas: (1) the laboratory detection of the autoantibody, (2) the detection of clinically significant red blood cell alloantibodies potentially masked by the autoantibodies, and (3) the selection and delivery of appropriate, although potentially incompatible, units. In addition, alternatives to red blood cell transfusion, specifically red blood cell substitutes, may also play an important role in the clinical treatment of these patients in the future. SUMMARY: In this article, we will review the most recent developments in the transfusion management of patients with autoimmune hemolytic anemia, specifically focusing on published articles between the period of May 2002 to April 2003. PMID- 14564174 TI - Risk and prevention of transfusion-related sepsis. AB - PURPOSE OF REVIEW: Transfusion-related sepsis is both the oldest recognized and most common transfusion-associated infectious risk. Despite an increased awareness and recognition of this problem, particularly with room-temperature stored platelets, strategies to prevent or reduce the occurrence of this problem have proved daunting. RECENT FINDINGS: With the recent FDA approval of culture methods for platelet bacterial testing and the promulgation of accreditation standards by the College of American Pathologists and American Association of Blood Banks to limit and detect platelet bacterial contamination, it is anticipated that the frequency of this problem will now begin to diminish. SUMMARY: As methods to reduce and/or inactivate pathogens emerge, it is hoped that transfusion-related sepsis will essentially disappear. PMID- 14564175 TI - Stem cell transplantation-harnessing of graft-versus-malignancy. AB - PURPOSE OF REVIEW: Allogeneic transplantation can cure a number of hematologic malignancies; however, the cost in morbidity and mortality is high. Much of the toxicity is a direct consequence of the intensity of the conditioning regimen. It has gradually been recognized that the conditioning regimen is important but not critical for the success of transplantation, particularly in the less aggressive hematologic malignancies. The graft-versus-malignancy effect, that is, the recognition of residual cancer cells by the T cells of the donor, is a critical component of the transplantation process. RECENT FINDINGS: This effect has been emphasized over high-dose therapy by using less intensive, and therefore less toxic conditioning regimens, thus allowing the graft-versus-malignancy effect to predominate. Reduced-intensity conditioning regimens have allowed the application of transplantation to older patients and to patients with underlying medical problems that preclude full-dose transplantation. SUMMARY: Although the long-term results of this type of approach have not yet been defined, it appears to be effective in diseases such as chronic myelogenous leukemia, chronic lymphocytic leukemia, and low-grade lymphomas that are not intrinsically very aggressive. Although the therapy appears to be valuable, concerns about delayed immune reconstitution and graft-versus-host disease remain. PMID- 14564176 TI - New approaches to invasive fungal infections. AB - Invasive fungal infections are typically difficult to prevent, diagnose, and treat; consequently, fungal infection-related mortality has become high, particularly among patients with prolonged neutropenia and severe graft-versus host disease. However, much recent progress has been made in understanding the immune response to fungi and the epidemiology of infection, and new diagnostic tests and antifungal drugs have become available. This review will describe these most recent developments with the goal of evaluating new approaches to prevent, diagnose, and treat invasive fungal infections. PMID- 14564177 TI - Hepatic veno-occlusive disease: pathogenesis, diagnosis and treatment. AB - Hepatic veno-occlusive disease (VOD) is one of the most serious complications following hematopoietic stem cell transplantation (SCT) and is associated with a very high mortality in its severe form. This review outlines the pathogenesis and clinical features of VOD, with an emphasis on endothelial cell injury and risk factors. The current status and future directions of research for both prophylaxis and treatment are also discussed. PMID- 14564178 TI - Acute lymphocytic leukemia: role of hematopoietic stem cell transplantation in current management. AB - PURPOSE OF REVIEW: It is still unclear what the best postremission therapy is for young adults with acute lymphocytic leukemia (ALL). Encouraging results, supporting the superiority of allogeneic stem cell transplantation in CR1 compared with chemotherapy/ allogeneic stem cell transplantation (ASCT), are coming from the ongoing international prospective MRC UKALL XII /ECOG 2993 study. However, the transplant-related morbidity and mortality remain high. RECENT FINDINGS: The detection of persistent minimal residual disease (MRD) after induction and consolidation therapy is being increasingly used to identify high risk patients for whom allogeneic transplant may offer the only curative option. SUMMARY: The ultimate aim is to determine whether the absence of MRD pretransplant can be used as a surrogate for low-risk adult ALL that can be cured without allogeneic transplantation. PMID- 14564179 TI - To cut or not to cut: that is the question. PMID- 14564180 TI - Duodenal cancer: endoscopic or surgical resection? PMID- 14564181 TI - Update on the genetics of inflammatory bowel disease. AB - There is a general consensus that interplay of genetic and environmental factors leads to an overactive mucosal immune response, which mediates the tissue damage in inflammatory bowel disease. Ethnic aggregation of inflammatory bowel disease (particularly, increased incidence and prevalence in the Ashkenazim), familial aggregation of inflammatory bowel disease, and greater concordance for inflammatory bowel disease in monozygotic twins than dizygotic twins are 3 lines of evidence for a central role of genetic factors in the pathogenesis. The genetics of inflammatory bowel disease cannot be explained by simple Mendelian genetics; it is characterized by incomplete penetrance, multiple susceptibility loci and genetic heterogeneity. Unraveling the complex genetics of inflammatory bowel disease is a daunting challenge, but the perseverance of inflammatory bowel disease gene hunters has produced commendable results in recent years. Since 1996, the field of inflammatory bowel disease genetics has progressed from publication of the first systematic genome searches for inflammatory bowel disease susceptibility loci to the identification of Crohn disease-associated genetic variants in CARD15/NOD2. Strategies for finding additional inflammatory bowel disease genes include taking advantage of the greater resolution and power of linkage disequilibrium mapping, mapping by admixture disequilibrium in African American and Hispanic-American populations, stratifying genetic analyses by genotypes at known inflammatory bowel disease loci, and refining inflammatory bowel disease phenotypes to reduce genetic heterogeneity and simplify the search for additional inflammatory bowel disease genes. PMID- 14564182 TI - Percutaneous endoscopic gastrostomy placement without a skin incision: results of a prospective pilot study. AB - Percutaneous endoscopic gastrostomy (PEG) tube placement may result in substantial skin site complications. Standard PEG (SPEG) placement involves performing a skin incision, which in our experience is not necessary. In a prospective pilot study, we examined the need for this incision with respect to placement success and stomal complications. Twenty consecutive patients underwent 20 Fr PEG tube placement by a modified technique, involving skin incision omission (IOPEG). Stoma evaluations were performed at 2 and 7 days after placement to record infection and bleeding scores. Mean infection scores were compared with published historical SPEG data obtained from this institution using the identical scoring system. In 12 cases, a digital force gauge measured peak pull-forces (pounds) required to pull the IOPEG through the abdominal wall and were compared with published SPEG pull-force data. Successful IOPEG placement was achieved in 20/20 (100%). No infectious or bleeding complications occurred. Mean 2 and 7-day infection scores were significantly lower than historical SPEG scores. Mean pull-forces were greater than published SPEG forces. A skin incision is not required for successful PEG placement. Incision omission likely increases PEG pull-forces, but may possibly lead to reduced infection complications. PMID- 14564183 TI - Malignant tumors of the small intestine. AB - SUMMARY: Malignant tumors of the small bowel are unusual and account for only 1% to 5% of all gastrointestinal tract malignancies. Thirteen cases of malignant tumors of the small bowel identified at the VAMC in Puerto Rico from January 1999 to September 2001 and a review of the literature are presented. The mean age of our cases was 67 (range: 45-78). Vague abdominal pain, nausea, vomiting, and melena were the most frequently reported symptoms. The average time from first symptoms to diagnosis was 3.2 months. A positive test for occult blood or hypochromic microcytic anemia was invariably present. Hyperbilirubinemia and increased alkaline phosphatase were warning signs that allowed earlier diagnosis in patients with duodenal tumors. Fifty-four percent of the lesions were detected by endoscopic examination while 46% relied on radiographic studies. Eleven had carcinomas and two malignant gastrointestinal stromal tumors. Neither carcinoids nor lymphomas were identified. Our report of thirteen cases of malignant small bowel tumors is unusual and exhibits differences with the previously reported data. The cases were identified in a limited Hispanic population in a short period of time. Aggressive evaluation and a high suspicion of these malignancies should be entertained whenever subtle symptoms and unexplained gastrointestinal blood loss are assessed. PMID- 14564184 TI - Clinicopathologic features and endoscopic resection of early primary nonampullary duodenal carcinoma. AB - Early primary nonampullary duodenal carcinoma is an extremely rare disease with poorly defined clinicopathologic features; early detection of this carcinoma is not common. To clarify the clinicopathologic characteristics of early primary nonampullary duodenal carcinoma and retrospectively analyze methods of treatment. Seventeen early duodenal carcinomas identified between 1994 and 2001 in 15 patients were studied. Lesions were divided into 2 groups: sporadic carcinoma (10 cases in 10 patients) and familial adenomatous polyposis associated carcinoma (7 cases in 5 patients). Clinicopathologic features and methods of treatment were compared between groups. The mean age of patients with sporadic carcinoma (63.8 years) was significantly higher than that of patients with FAP-associated carcinoma (34.9 years). The incidence of sporadic carcinoma was significantly higher in men that in women (M:F ratio 9:1); the difference between sexes in the incidence of FAP-associated carcinoma (1:4) was not significant. There was no significant difference between both groups in relation to tumor size, location, gross appearance, or histopathology. Thirteen of the duodenal tumors were treated by endoscopic mucosal resection (EMR), two by polypectomy, and two by open surgery. Complications were encountered in 1 of 15 cases (6%); local bleeding occurred after one EMR, but hemostasis was achieved endoscopically. The mean follow-up period for all patients was 51.7 months. No patients experienced recurrence after resection. The significant differences between patients with sporadic and those with early FAP-associated duodenal carcinoma were in age and sex. Endoscopic resection appears to be a safe and efficient treatment of carefully selected patients with early primary nonampullary duodenal carcinoma. PMID- 14564185 TI - Lack of usefulness of anti-transglutaminase antibodies in assessing histologic recovery after gluten-free diet in celiac disease. AB - Gluten-free diet (GFD) plays a key role in the treatment of celiac disease (CD), but it is difficult to evaluate the effect of GFD on the improvement of villous architecture using sensitive, non-invasive tests. Aim of this study is to evaluate anti-transglutaminase (tTG) antibodies in the follow-up of CD to detect histologic recovery. We studied 42 consecutive patients with CD. In all the patients anti-tTG antibodies (evaluated by the enzyme linked immunosorbent assay method) and EGDscopy with multiple bioptic samples before GFD and then 6, 12, and 18 months after GFD were evaluated. For comparison, a sorbitol H2-breath test (H2 BT) and anti-endomysium (EMA) antibodies test were carried out concomitantly. Anti-tTG results were positive in 36 of 42 patients before GFD (80.95%), while they were positive in 11 of 34 (32.35%), 1 of 17 (5.88%), and 0 of 6 (0%) of patients with a persistence in histologic lesions 6, 12, and 18 months of GFD respectively, without any correlation with persistence of histologic lesions (P = NS). Also EMA failed to show correlation with improvement of histologic lesions. They were positive in 31 of 42 patients before GFD (73.80%), while they were positive in 18 of 34 (52.94%), 3 of 17 (17.64%), and 0 of 6 (0%) cases 6, 12, and 18 months of GFD respectively (P = NS). Regarding sorbitol H2-BT, it was positive in 40 of 42 (95.24%) patients before GFD, while it was positive in 31 of 34 (91.17%), 13 of 17 (76.47%), and 4 of 6 (50%) of patients with a persistence in histologic lesions 6, 12, and then 18 months after GFD starting (see Fig. 2, infra). So, anti-tTG and EMA were ineffective in assessing the histologic recovery at each follow-up visit (P = NS), while sorbitol H2-BT seems more effective than anti-tTG and EMA in this field (P < 0.0001 sorbitol H2-BT versus anti-tTG and versus EMA at 18 months after gluten withdrawal). Thirty-eight of 42 (90.47%) patients adhered to a strict GFD. Four patients were found to have occasional dietary transgression, and in all we noted a progressive decreasing of anti-tTG after 6 months of GFD and negative anti-tTG after 12 months of GFD, but sorbitol H2-BT persisted being positive during the entire follow-up. Intestinal damage persisted during the follow-up, despite anti-tTG and EMA negativity, and worsened in the presence of dietary lapses. Anti-tTG does not seem effective to assess histologic recovery in the follow-up of celiac patients after they have started GFD due to its poor correlation with histologic damage. PMID- 14564186 TI - Argon plasma coagulation of hemorrhagic solitary rectal ulcer syndrome. AB - Solitary ulcer syndrome (SUS) is a rare disorder that may provoke hematochezia. Argon plasma coagulation (APC) is used in a wide range of gastrointestinal bleeding. We experienced APC in a patient with a bleeding gigantic SUS: a 64-year old woman who developed a SUS at 60. After 3 years, recurrent hematochezia, secondary anemia, and rectal pain occurred. Endoscopy revealed a large rectal bleeding ulcer. Moreover, the pain led the patient to assume analgesics. These conditions stimulated us to treat this ulcer with APC within 4 sessions; each session spaced out at 30-day intervals. The patient experienced and maintained the following benefits: (1) resolution of bleeding and secondary anemia after the first session, (2) reduction of ulcer depth, disappearance of pain and analgesic withdrawal at the end of the cycle, (3) almost complete endoscopic healing of the ulcer after 9 months of follow-up. This experience suggests that APC may represent a therapeutic approach for bleeding SUS even if controlled studies are necessary before recommending it as acceptable treatment. PMID- 14564187 TI - Treatment considerations in patients with hepatitis C and cirrhosis. AB - Patients with cirrhosis due to hepatitis C have a high chance of dying from progressive liver disease and thus have much to gain from successful antiviral therapy. The highest sustained virologic responses in patients with cirrhosis have been achieved using pegylated interferon alfa plus Ribavirin; 43% or more remain with undetectable virus 6 months after the cessation of 48 weeks of treatment. In those who achieve a sustained virologic response, the degree of fibrosis is less as judged on posttreatment liver biopsy; cirrhosis may even regress. In those individuals with cirrhosis who achieve a sustained virologic response, the risk of developing hepatocellular carcinoma is significantly reduced and it is likely that their chance of developing liver failure is less. Patients who do not achieve sustained virologic response can still show histologic improvement as demonstrated on liver biopsy posttherapy as compared to baseline. Patients with compensated cirrhosis can benefit from therapy while those who are decompensated are prone to more safety issues. Thus, individuals with any evidence of hepatic decompensation should generally not be given interferon-based antiviral therapy, but treatment should be encouraged for those whose status is Child Class A. PMID- 14564189 TI - Five days of ceftriaxone to treat culture negative neutrocytic ascites in cirrhotic patients. AB - The goal of this study is to establish whether 5 days of ceftriaxone treatment was sufficient to cure culture-negative neutrocytic ascites in cirrhotic patients. We studied 50 cirrhotic patients with culture-negative neutrocytic ascites. All were treated with ceftriaxone, 1.0 g IV, twice a day for 5 days. A control paracentesis was performed 48 hours after starting the therapy to assess response to the treatment. A total of 17 demographic, clinical, and laboratory variables were recorded in all cases on the day of diagnosis of CNNA. The mean age of the patients was 57.7 +/- 13.2 years. Thirty-two patients were males and 18 females. The etiology of cirrhosis was hepatitis C virus in 20 patients (40%), hepatitis B virus in 16 patients (32%), cryptogenic in 13 patients (26%), and alcohol abuse in 1 patient (2%). Eighty percent of the patients were in Child Pugh Class C. Resolution rate of culture-negative neutrocytic ascites on day 5 of treatment was 78%. Hospital mortality in cirrhotic patients with culture negative neutrocytic ascites was 4%. Statistical analysis showed that none of the 13 selected variables as covariates significantly related with the resolution of culture-negative neutrocytic ascites. Five days of ceftriaxone treatment is an adequate therapy for culture-negative neutrocytic ascites. PMID- 14564188 TI - Association of nonalcoholic fatty liver disease with insulin resistance: is OGTT indicated in nonalcoholic fatty liver disease? AB - Nonalcoholic fatty liver disease (NAFLD) is frequently associated with type 2 diabetes mellitus, obesity, and dyslipidemia. We tested the hypothesis that there may be an association between NAFLD and insulin resistance (IR); and its correlation with glucose tolerance status of subjects who aren't known patients with diabetes. One hundred and seventy-six consecutive patients with elevated serum aminotransferase levels and bright liver were evaluated. Sixty-two patients were excluded from the study. Age, gender, height, weight, body mass index, waist circumferences, and family history of diabetes were recorded. Fasting plasma glucose, insulin, lipid profile were measured. A standard oral glucose tolerance test (OGTT) was performed and the index of IR was calculated according to the HOMA method. Patients with a normal glucose tolerance formed group 1 (64 patients) and patients with impaired or diabetic glucose tolerance group 2 (50 patients). Age, female sex, family history of type 2 diabetes, fasting insulin, fasting plasma glucose and HOMA-R index were statistically significantly different between the groups. Although the subjects in the study are not known patients with diabetes, the prevalence of impaired or diabetic glucose tolerance was prominent. In conclusion, performing OGTT in cases with nonalcoholic fatty liver disease may be useful for early screening of diabetes mellitus. PMID- 14564190 TI - Pilot study of the relationship between histologic progression and hepatic iron concentration in chronic hepatitis C. AB - Hepatic iron deposition is common in patients with chronic hepatitis C (HCV) and may play a role in progression of liver disease. This pilot study examines the relationship between hepatic iron concentration (HIC) and histologic progression over time in patients with HCV. HIC was retrospectively measured in 14 patients with HCV who had 2 serial liver biopsies prior to the era of interferon therapy. The mean interval between biopsies was 56 +/- 46 months. Mean Knodell score worsened between first and second biopsies (10.0 +/- 2.8 versus 12.4 +/- 3.3; P = 0.007). There was increased portal inflammation (3.2 +/- 0.4 versus 3.6 +/- 0.5; P = 0.028) and fibrosis (1.8 +/- 1.3 versus 2.7 +/- 1.2; P = 0.002), but no significant change in piecemeal necrosis or lobular degeneration. There was no significant change in HIC between first and second biopsy (P = 0.66). However, HIC was noted to increase significantly among patients with cirrhosis on initial biopsy or those who progressed to cirrhosis (P = 0.009). In this pilot study, histologic progression in patients with precirrhotic HCV was not associated with an increase in HIC, whereas hepatic iron accumulation was observed in 3 patients once cirrhosis was present. The interaction between progression of hepatitis C and iron deposition warrants further study. PMID- 14564191 TI - Transradial approach for transcatheter arterial chemoembolization in patients with hepatocellular carcinoma: comparison with conventional transfemoral approach. AB - We evaluated the clinical usefulness and safety of transradial approach for transcatheter arterial chemoembolization (TACE) in patients with hepatocellular carcinoma (HCC) compared with that of conventional transfemoral approach. The two groups (radial group, n = 177; femoral group, n = 150) of cases were retrospectively compared with regard to the successful rate of angiography or TACE, time required for catheterizaiton and complications. Hepatic angiography and TACE were completed in 174 (98.3%) of 177 cases in the radial group. There was no intergroup difference of time required for catheterization. Minor complications (dull pain, numbness) occurred in 8 (4.6%) patients in the radial group, and there were lower complications in the radial group compared to the femoral group. TACE by our new transradial approach was found to have therapeutic efficacy with lower complications comparable to that of the conventional transfemoral approach. PMID- 14564192 TI - Gene microarrays in transplantation. PMID- 14564193 TI - The implications of the ADEMEX study for the peritoneal dialysis prescription: the role of small solute clearance versus salt and water removal. AB - PURPOSE OF REVIEW: This review examines the results of the ADEMEX (Adequacy of Peritoneal Dialysis in Mexico) study in the context of other recent advances in peritoneal dialysis, and assesses the implication of this new knowledge for the optimal peritoneal dialysis prescription. RECENT FINDINGS: The prospective randomized controlled ADEMEX study demonstrated no survival advantage of an increased dose of peritoneal small molecule clearance delivered by chronic ambulatory peritoneal dialysis. Coincident with this finding, there has been increasing awareness that many peritoneal dialysis patients are volume expanded, and that there are adverse cardiovascular consequences to this chronic overhydration. As a result there has been a shift away from interest in peritoneal small solute clearance with renewed interest in peritoneal removal of salt and water. There is also increasing evidence of the importance of residual renal function in maintaining euvolemia and as a prognostic indicator for survival. SUMMARY: The ADEMEX study and subsequent investigations have changed the way we perceive the optimal peritoneal dialysis prescription. This has resulted in de-emphasis of peritoneal small molecule clearance and increased emphasis on clinical assessment of dialysis adequacy, preservation of residual renal function, and optimization of salt and water removal. PMID- 14564194 TI - The HEMO study - where do we go from here? AB - PURPOSE OF REVIEW: The HEMO study is a randomized clinical trial using a 2x2 factorial design to assign patients to a standard or high dose of dialysis and to a low flux or a high flux dialyzer. This study is the largest, most comprehensive, randomized clinical trial ever performed in the maintenance hemodialysis population. This review analyzes the results of the study and discusses how the HEMO study results affect efforts to lower morbidity and mortality in the hemodialysis population. RECENT FINDINGS: The primary outcome of the HEMO study was death from any cause. This outcome was not significantly influenced by treatment assignment for the dose of dialysis or for the flux of the dialysis membranes used. The main secondary outcomes also did not differ significantly in either the dose groups or the flux groups. SUMMARY: The results of the HEMO study support current clinical practice guidelines for the delivery of thrice-weekly dialysis, but the results do not support conventional attempts to lower the high morbidity and mortality in hemodialysis patients. Current efforts are being focused on increasing dialysis time and/or frequency, improving phosphate control, and lowering traditional and nontraditional risk factors for adverse cardiovascular events in this patient population. PMID- 14564195 TI - Oxidative stress in uremia. AB - PURPOSE OF REVIEW: Oxidative stress has been described as 'a disturbance in the prooxidant-antioxidant balance in favor of the former, leading to potential damage. In uremic patients, an increase in oxidative stress may occur because of the loss of residual renal function, and may be exacerbated by dialysis. This review will focus on the emerging biochemical evidence of an increase in oxidative stress in uremic patients, the relationship with renal replacement therapy, and the potential linkages to acute-phase inflammation, malnutrition, and adverse cardiovascular outcomes in uremic patients. RECENT FINDINGS: Many studies from multiple research laboratories around the world have recently utilized in-vivo biomarkers to describe increased oxidative stress in uremic patients. An emerging literature suggests that there are links between an increase in oxidative stress, endothelial dysfunction, an increase in acute-phase inflammation, and an accelerated risk of cardiovascular complications in dialysis patients. Additional uremia-associated metabolic abnormalities, including hyperhomocysteinemia, intravenous iron exposure, and biocompatibility changes related to dialysis, may contribute to an increase in oxidative stress. Finally, two well-conducted pilot clinical randomized trials have suggested that antioxidant therapy may have efficacy in reducing cardiovascular events in uremic patients. SUMMARY: The implications of the findings of a generalized increase in oxidative stress associated with uremia have led to the suggestion that antioxidative therapy may be efficacious in reducing cardiovascular complications. Pilot studies have suggested potential efficacy for this approach. However, further large-scale randomized clinical trials will be required to establish a compelling, evidence-based approach to the use of antioxidants in patients with uremia. PMID- 14564197 TI - Pharmacogenomics of immunosuppressive drug metabolism. AB - PURPOSE OF REVIEW: The immunosuppressants are potent and toxic drugs with narrow therapeutic ranges. The pharmacokinetic variability of these drugs has made establishing appropriate dosing difficult. Currently, therapeutic drug monitoring is an important adjunct to achieving the precarious balance between efficacy and toxicity. However, pharmacogenomic analysis has the potential to improve dosing strategies. Several of the drugs in this category are metabolized through complex pathways, which have the potential to be affected by genetic traits. The current literature has addressed several genes and polymorphisms in relation to these drugs. RECENT FINDINGS: Polymorphisms related to the coding of P-glycoprotein (coded by the MDR-1 gene) and cytochrome P450 3A enzymes have been the main focus of research. These gene products are involved in regulating the absorption and metabolism of the principal immunosuppessants. Two polymorphisms (C3435T and G2677[A/T]) of the MDR-1 gene have been shown to influence the bioavailability and toxicity of tacrolimus and cyclosporin. Phase I metabolism of these drugs has been shown to be affected by two polymorphisms (CYP3A5*1 and CYP3AP1*1), related to cytochrome P450 3A5 expression, rather than cytochrome P450 3A4. SUMMARY: The current literature has shown disparity as to which are the most important polymorphisms affecting the metabolism of immunosuppressants. Although pharmacogenomics has the potential to allow improvements in devising optimal dosing regimes, it has not yet offered any definitive solutions to the problems of dosing, because the process of elucidating the complex influence of genetics on drug metabolism is only starting to be unravelled. PMID- 14564196 TI - Polyomavirus nephropathy: morphology, pathophysiology, and clinical management. AB - PURPOSE OF REVIEW: Viral nephropathies, particularly those caused by polyomaviruses of the BK-virus strain, are serious complications following renal transplantation. The review will highlight the morphological, pathophysiological and clinical aspects of BK-virus nephropathy. New patient management strategies are discussed. RECENT FINDINGS: Immunosuppression with tacrolimus and mycophenolate-mofetil promotes the activation of latent BK-virus in the urinary tract and increases the odds ratio for developing BK-virus nephropathy significantly. A productive infection with BK-viruses shows viral replication in tubular epithelial cells and acute tubular injury. BK-virus nephropathy can be further complicated by concurrent acute rejection episodes contributing to graft demise. Risk assessment after transplantation and patient management during ongoing viral nephropathy have undergone revision by the introduction of real time quantitative polymerase chain reaction techniques measuring BK-virus genome load fluctuations in the serum. Treatment strategies for BK-virus nephropathy include not only low-dose immunosuppression but also drugs with antiviral effects: cidofovir and leflunomide. Transient anti-rejection therapy, including anti-lymphocytic preparations, is a therapeutic option in cases of BK-virus nephropathy and concurrent acute rejection. Recent advances in patient management strategies have resulted in markedly improved graft survival. In cases of graft loss due to BK-virus nephropathy, re-transplantation should be considered. SUMMARY: BK-virus nephropathy is a significant complication following renal transplantation. Recent advances have improved our understanding of the morphological changes, potential risk factors and patient management strategies would be optimized. The availability of quantitative viral load measurements now offers the opportunity for a more accurate and timely clinical intervention. PMID- 14564198 TI - Proteinuria: new information from an old friend. PMID- 14564199 TI - Diagnostic and prognostic significance of anti-C1q antibodies in systemic lupus erythematosus. AB - PURPOSE OF REVIEW: The presence of a wide variety of autoantibodies is a characteristic finding in systemic lupus erythematosus. Autoantibodies against nuclear proteins, such as anti-nuclear and anti-double-stranded DNA antibodies, are used as diagnostic markers in systemic lupus erythematosus. Renal involvement is frequently found in systemic lupus erythematosus and is an important risk factor for death. Therefore, markers for the diagnosis and follow-up of nephritis are very important. Anti-C1q autoantibodies are strongly associated with renal involvement in systemic lupus erythematosus. This study will review recent findings on the pathogenic role and clinical importance of anti-C1q antibodies in lupus nephritis. RECENT FINDINGS: Recent clinical studies have clearly emphasized the diagnostic relevance of anti-C1q autoantibody levels in patients with lupus nephritis. With a possible negative predictive value of 100%, anti-C1q autoantibodies are the only exclusive antibodies associated with the involvement of a single organ in systemic lupus erythematosus. Next to the clinical findings, the pathogenic significance of anti-C1q antibodies has been shown in an animal model. The deposition of autologous C1q in healthy glomeruli of mice after the infusion of anti-C1q antibodies induces moderate tissue damage. SUMMARY: The latest insight into the pathogenesis of anti-C1q autoantibodies in the development of lupus nephritis and the recently demonstrated clinical importance of anti-C1q autoantibodies for the diagnosis of lupus nephritis support the value of further investigations. New diagnostic methods for the detection of anti-C1q and an accurate follow-up of antibody levels might be of use in clinical practice. PMID- 14564200 TI - The clinical art and science of urine microscopy. AB - PURPOSE OF REVIEW: The examination of urine sediment is a diagnostic test which is frequently neglected by nephrologists. With this review the authors wanted to demonstrate that it can provide useful and relevant information in a wide spectrum of clinical situations. RECENT FINDINGS: The authors reviewed the main contributions dealing with urine sediment examination, published in international journals in the period from January 2002 to April 2003. After a section on methodological aspects, they described the importance of urine sediment examination in various diseases of the urinary tract. These included bladder B lymphoma, systemic histoplasmosis, urate nephropathy, Fabry disease, myeloma cast nephropathy, giant cell arteritis, and lupus nephritis. The significance of 'decoy cells' in the urine as a marker of polyomavirus BK reactivation was also discussed, both in renal transplantation and other conditions such as solitary pancreas transplantation, chronic lymphatic leukaemia, and HIV infection. In the section devoted to urine sediment changes caused by drugs the authors dealt with leukocyturia induced by indinavir, and crystalluria, which can follow amoxycillin and acyclovir administration. Finally, they reported on the utility and limits of flow cytometry for the automated analysis of urine sediments. SUMMARY: The review of the recent literature on urine sediment examination shows that this test has important clinical implications in a large spectrum of diseases. Therefore, it should be more widely used by nephrologists. PMID- 14564201 TI - Developments in online monitoring of haemodialysis patients: towards global assessment of dialysis adequacy. AB - PURPOSE OF REVIEW: Online monitoring of haemodialysis provides more detailed and more immediate measurement of parameters currently assessed during haemodialysis. It can also assess novel variables that are not routinely measured and could intensively monitor the haemodynamic response to dialysis. In an era of expanding numbers of increasingly dependent patients on dialysis, the ability to provide intensive and extensive monitoring of haemodialysis is a boon. The technology available to do this is rapidly becoming more sophisticated, more widely available and more diverse than ever before. RECENT FINDINGS: Adequacy of urea removal is of crucial importance to mortality on haemodialysis, and online monitoring of urea removal is now achievable using a number of different methods. Some of these methods also measure sodium flux, allowing precise monitoring of sodium balance. Haemodynamic disturbance can be assessed using relative blood volume monitoring equipment, or more directly with non-invasive pulse-wave analysis giving a continuous blood pressure reading. The attributes of each of the separate technologies involved are examined. SUMMARY: Online monitoring of a variety of pertinent parameters during haemodialysis provides an array of tools that are steadily being incorporated into routine clinical practice. Research is also benefiting from novel insights into the process of haemodialysis provided by this technology. PMID- 14564202 TI - Urinary protein and enzyme excretion as markers of tubular damage. AB - PURPOSE OF REVIEW: In this review we intend to evaluate the clinical usefulness of measurement of urinary excretion of microproteins and tubular enzymes. RECENT FINDINGS: Studies of patients with acute tubular necrosis showed that the excretion of some well known enzymes and of new markers such as kidney injury molecule-1 has higher value than classic renal function tests for the early detection of the disease as well as for the prediction of the development of overt acute renal failure. Tubular dysfunction may be detected by measurement of the excretion of tubular enzymes and microproteins not only in workers chronically exposed to heavy metals but also in population exposed to low environmental level below the limits set by the WHO. In chronic glomerulonephritis the excretion of alpha1-microglobulin is significantly associated with the extent of tubulo-interstitial damage and predicts outcome better than proteinuria/day; in the same diseases NAG excretion is significantly dependent on immunoglobulin G and alpha1-microglobulin excretion and predicts remission, progression and response to therapy. SUMMARY: The measurement of urinary excretion of tubular enzymes and microproteins may be clinically useful in several different kidney diseases (such as acute tubular necrosis, glomerulonephritis) both for diagnosis and prediction of fractional outcome. PMID- 14564203 TI - Recent insights from studies using ambulatory blood pressure monitoring in patients with renal disease. AB - PURPOSE OF REVIEW: To identify and evaluate recent (2000-2003) published studies employing ambulatory blood pressure monitoring in patients with chronic renal failure, on dialysis, and after renal transplantation. RECENT FINDINGS: We discuss several studies that have employed ambulatory blood pressure monitoring to refine the analysis of the link between blood pressure levels, and diurnal alterations, and end-organ damage or patient survival. There is now some evidence that an abnormal diurnal blood pressure profile, although intrinsically not a very reproducible label, has predictive value for patient survival, and that the non-dipping phenomenon is linked to a high incidence of cardiovascular disease and autonomic dysfunction. SUMMARY: Ambulatory blood pressure monitoring remains an important adjunct to the comprehensive cardiovascular evaluation of patients with chronic, end-stage renal failure or after renal transplantation. PMID- 14564204 TI - Radionuclide renography: a review. AB - PURPOSE OF REVIEW: Radionuclide renography has become standard practice but with controversial areas and variations in technique. This review looks at some recent studies to improve usefulness and to assess the current clinical indications for the use of dynamic radionuclide renography and considers some of the recent publications that have contributed to the subject during the past year. RECENT FINDINGS: Two important papers attempting to standardize the diuretic renogram and provide normal values for the MAG3 renogram are reviewed together with exercise renography in hypertension and used for the Captopril test. SUMMARY: Research into renography is not an active area at present but there have been a few important contributions which will improve the clinical value of radionuclide renography. PMID- 14564205 TI - Genetic screening in haemolytic uraemic syndrome. AB - PURPOSE OF REVIEW: Haemolytic uraemic syndrome (HUS) is a disease of diverse origin. The last year has witnessed the identification of a novel genetic marker of this disease, the description of the frequency of the factor H associated form of HUS in a registry of over 100 patients and a better understanding of the pathophysiology of the disease. RECENT FINDINGS: In patients with atypical HUS, heterozygous mutations in the gene coding for the soluble complement regulator factor H are reported and most of the mutations cluster in the C-terminal recognition domain of the protein. A novel genetic marker for HUS has also been identified. Mutations occurring in the gene of the von Willebrand factor cleaving protease, ADAMTS13, which were previously linked to thrombotic thrombocytopenic purpura have now been identified in HUS patients. The frequency of factor H associated HUS was established as 14% in a registry of German speaking countries and also 16 novel disease associated mutations were reported. The pathophysiology of factor H-associated HUS was analysed. Three analysed mutant proteins show normal complement regulatory activities but display defective recognition functions: reduced binding to surface attached C3b, to heparin/polyanions and to endothelial cells. SUMMARY: The identification of effector molecules of the complement as well as the coagulation cascade as disease associated molecules indicate a regulatory protein network, which maintains integrity of endothelial cells during stress or infection. Defining the individual components and how their functional interaction causes microangiopathies will identify additional disease markers and will allow the design of proper diagnostic and therapeutic approaches. PMID- 14564206 TI - Craniosynostosis associated with intracranial calcification: a novel recessive syndrome. AB - We report three siblings who were variably affected by craniosynostosis, calcification of the basal ganglia, and mild facial dysmorphism comprising prominent eyes and a prominent nasal bridge. The children are of normal intelligence and have no limb abnormalities. Their parents are first cousins and are phenotypically normal. We propose that this combination of clinical findings represents a recognizable, autosomal recessive craniosynostosis syndrome. PMID- 14564207 TI - Holoprosencephaly, bilateral cleft lip and palate and ectrodactyly: another case and follow up. AB - We describe a male patient with lobar holoprosencephaly, ectrodactyly, and cleft lip/palate, a syndrome which has been seen previously in only six patients. In addition, our patient developed hypernatraemia, which has been described in three patients before. PMID- 14564208 TI - Further delineation of Hennekam syndrome. AB - We report four children from four inbred Arab families with varying manifestations of Hennekam syndrome and additional features that have not been previously reported. These include abnormalities of the middle ear, anomalous pulmonary venous drainage, interrupted inferior vena cava, polysplenia, crossed renal ectopia, median position of the liver and multiple cavernous haemangiomas. In addition, in one case lymphoedema was absent and oedema due to hypoproteinaemia appeared at 6 years of age. Since anomalies of the veins and the consequent developmental abnormalities of the lymphatics might lead to alterations in the fluid balance of the embryo, we hypothesize that altered fluid dynamics due to defective vascular and lymphatic development might disrupt critical events in craniofacial morphogenesis resulting in Hennekam syndrome. PMID- 14564209 TI - Frontonasal dysplasia, Poland anomaly and unilateral hypoplasia of lower limb: report on a male patient. AB - We report a 13-year-old boy with an apparently previously undescribed combination of findings including facial features of frontonasal dysplasia, Poland anomaly, and contralateral hypoplasia of the lower limb. Clinical correlations among these features are discussed. PMID- 14564210 TI - Nicolaides-Baraitser syndrome: confirmatory report of a syndrome with sparse hair, mental retardation, and short stature and metacarpals. AB - In 1993, Nicolaides and Baraitser reported a new condition consisting of short stature, hypotrichosis, brachydactyly with cone-shaped epiphyses, epilepsy and severe mental delay. We report here two unrelated patients with identical features, thus confirming the status of this rare disorder of unknown aetiology. PMID- 14564211 TI - An Irish three-generation family of Cornelia de Lange syndrome displaying autosomal dominant inheritance. AB - The existence of familial de Lange syndrome has been documented in sibs and in parent-child families, but the inheritance pattern continues to be the cause of much debate. We describe a classically affected neonate with de Lange syndrome, an affected mother and probably affected maternal grandmother. These cases show evidence for a dominantly inherited syndrome with a de Lange phenotype. PMID- 14564212 TI - Macrocephaly and sclerosis of the tubular bones in an isolated patient: a mild case of craniodiaphyseal dysplasia? AB - We report a 56-year-old woman, mainly suffering from painful legs and the inability to run. Radiologically, marked sclerosis and hyperostosis of the skull bones is present resulting in macrocephaly. Most tubular bones of the limbs, as well as the clavicles, are affected by sclerosis. By mutation analysis of the TGFB1, SOST and LRP5 genes, we were able to exclude the diagnoses of Camurati Engelmann disease, Van Buchem disease, sclerosteosis, high-bone-mass trait and endosteal hyperostosis (Worth type). We believe this patient represents one of the very few examples of adult craniodiaphyseal dysplasia with a mild form of the disease and moderate facial changes. PMID- 14564213 TI - A female infant with duplication of chromosome 2q33 to 2q37.3. AB - We report a 13-month-old female child with a de-novo inverted duplication of chromosome 2q extending from 2q33 to 2q37.3. She had microcephaly and craniofacial dysmorphism compatible with previously reported cases with overlapping duplications of chromosome 2q. Although a facial phenotype for pure partial trisomy 2q3 has been described, some controversy still exists regarding possible band specificity for the facial findings. We consider this child provides further evidence for a recognizable facial appearance associated with duplication of chromosome band 2q33 to 2q37.3. Other clinical features found with duplication for chromosome 2q3 have been variable and we provide a summary of the findings in previously reported cases. PMID- 14564214 TI - Partial distal 5p trisomy resulting from paternal translocation (5;15)(p15.1;p13) in a boy with no mental retardation. AB - We report a boy with partial distal 5p15.1-->pter trisomy and normal development. We compared the clinical findings in our patient with those previously reported of the same 5p duplicated region. Several cases of autosomal duplications and normal development have been described. The present case is another example of a chromosomal anomaly with little, if any, phenotypic effect without mental retardation. PMID- 14564215 TI - Unbalanced cryptic translocation der(14)t(9;14)(q34.3;q32.33) identified by subtelomeric FISH. AB - We investigated a girl with dysmorphic features and moderate developmental delay by subtelomeric FISH (fluorescence in-situ hybridization). We found an unbalanced cryptic translocation, t(9;14)(q34.3;q32.33), resulting in a subtelomeric deletion of 14q and duplication of 9q deriving from a balanced translocation in the mother. A review of the literature suggests that the phenotype of our case is related to the 14 qter deletion, without signs of concomitant partial trisomy 9. The case reinforces the value of subtelomeric screening for genetic counselling. PMID- 14564216 TI - Thyroid hypoplasia of the left lobe in two girls affected by Williams syndrome. AB - Williams syndrome is a well-recognized disorder, having an incidence of 1 in 20,000 live births. However, thyroid function in these patients is rarely studied. This paper reports thyroid hypoplasia of the left lobe in two girls affected by Williams syndrome, suggesting that it may be a feature of this syndrome. PMID- 14564217 TI - The appearance of the feet in Pfeiffer syndrome caused by FGFR1 P252R mutation. AB - Patients affected by Pfeiffer syndrome generally present with syndromic craniosynostosis and typical limb defects including broad thumbs, wide halluces with varus deformity, toe syndactyly and sometimes elbow ankylosis. This autosomal dominant condition can be caused by mutations in either fibroblast growth factor receptor gene type 1 or 2 (FGFR1 or FGFR2). We report four new affected families showing an FGFR1 P252R mutation and emphasize the characteristic malformations of the feet in this form of Pfeiffer syndrome. In one family this was the only abnormality. PMID- 14564218 TI - Type 1 Arnold-Chiari malformation and Noonan syndrome. A new diagnostic feature? AB - Noonan syndrome is a clinically and genetically heterogeneous genetic condition. Arnold-Chiari malformation has been previously reported in three cases of Noonan syndrome. We describe a fourth case with this association. We suggest that brain and cervical spine MRI should be performed if neurological symptoms are present. PMID- 14564219 TI - Diaphragmatic hernia and preaxial polydactyly in spondylothoracic dysplasia. AB - We report on two cases (male twins), and a female sib terminated at 20 weeks, whose autopsy revealed features of spondylothoracic dysplasia (STD) and also a diaphragmatic hernia and preaxial polydactyly. We present the findings and review STD and the closely related spondylocostal dysostosis. On the basis of the discussion we suggest that our cases are possibly the first report of preaxial polydactyly in spondylothoracic dysplasia and that STD and spondylocostal dysostosis may be allelic. PMID- 14564220 TI - Catel-Manzke syndrome without cleft palate: a case report. AB - We report a boy who presented with mild facial dysmorphism, pectus excavatum, bilateral brachydactyly and ulnar deviation of both index fingers. Radiological examination of the hands showed abnormal proximal phalanges of the second digits. This case differs from the classical Catel-Manzke syndrome in having isolated hand malformations without any other malformations such as cleft palate or cardiac abnormalities. PMID- 14564221 TI - Unusual combination of limb malformations in the same patient: brachydactyly with syndactyly and postaxial polydactyly of the hands and postaxial oligodactyly of the feet. AB - We report on a case with polydactyly, syndactyly and brachydactyly of the hands and oligodactyly of the feet, but no other anomalies and normal chromosome analysis. We compared the findings in our case with those of brachydactyly B, Fuhrmann syndrome and Haas-type syndactyly. However, it was not possible to suggest a syndrome diagnosis in the present case. PMID- 14564222 TI - A case of dextrocardia, radial ray malformation and renal anomaly. AB - A 12-year-old boy is described with bilateral radial club hands, scoliosis, hypospadias, isolated dextrocardia, hypoplastic ribs, an ectopic kidney and spina bifida occulta. Although some of the clinical features of this patient are seen in VATER association and sacrococcygeal dysgenesis, the presence of dextrocardia, facial dysmorphism, radial, renal and vertebral anomalies preclude these diagnoses. PMID- 14564223 TI - Cardiac natriuretic peptides:a proteomic window to cardiac function and clinical management. AB - Congestive heart failure (CHF) is a leading cause of adult hospitalization in the United States, and despite advancements in treatment, the disease remains a major clinical challenge. The chief symptom of CHF is dyspnea, but in the urgent-care setting, it is often difficult to distinguish between cardiac and pulmonary causes of this symptom. B-type natriuretic peptide (BNP) is mainly synthesized, stored, and released in the ventricular myocardium and is strongly induced during ventricular-wall tension or stretch. It can be measured rapidly at the point of care and can be used to differentiate cardiac from pulmonary etiologies of dyspnea. In addition to its diagnostic utility, it also has prognostic value and may help guide the treatment of patients with CHF. Thus, it is likely that future algorithms incorporating BNP levels and other clinical indicators will become available to guide critical-care physicians in making management decisions for their CHF patients. PMID- 14564224 TI - Sorting out the evidence on natriuretic peptides. AB - B-type natriuretic peptide (BNP) is a cardiac neurohormone released as pre-proBNP and then enzymatically cleaved to the N-terminal-proBNP (NT-proBNP) and BNP upon ventricular myocyte stretch. Blood measurements of BNP and NT-proBNP have been used to identify patients with heart failure (HF). Clinical considerations for these tests include their half-lives in plasma, dependence on renal function for clearance, interpretation of their units of measure, and the rapid availability of the test results. The BNP assay is currently used as a diagnostic and prognostic aid in HF and as a prognostic marker in acute coronary syndromes (ACS). In general, a BNP level less than 100 pg/mL excludes acutely decompensated HF. In the absence of renal dysfunction, NT-proBNP has also been shown to be of diagnostic value in HF, related to HF severity, predictive of sudden death, and prognostic for death in ACS. This article will sort out the literature concerning the use of these peptides in a variety of clinical scenarios. PMID- 14564225 TI - Combining natriuretic peptides and necrosis markers in determining prognosis in heart failure. AB - Despite significant advances in medical therapy, patients with heart failure remain at increased risk of overall mortality, progressive ventricular dysfunction, and sudden cardiac death. Although a number of individual clinical and laboratory variables have been identified as being associated with increased mortality risk in heart failure, there remains a clear need for an integrated, accurate method of determining prognosis. Elevated plasma B-type natriuretic peptide (BNP) has been demonstrated to be a powerful marker for prognosis and risk stratification in the setting of heart failure. Patients with elevated BNP levels have been shown to be at significantly higher risk for heart failure admission or death, and higher BNP levels are associated with progressively worse prognosis. Although cardiac troponins are a well-established diagnostic and prognostic marker in acute coronary syndromes, emerging data suggest that cardiac troponins also provide independent prognostic information in heart failure. Detection of cardiac troponins in the serum of patients with heart failure has been shown to be associated with an impaired hemodynamic profile, progressive decline in left ventricular systolic function, and shortened survival. Combining a marker of myocyte injury-cardiac troponin-with BNP in a multimarker strategy appears to be a useful tool for improving risk assessment and triage in patients with heart failure. Heart failure patients with detectable cardiac troponin I and high BNP levels have been shown to have a 12-fold increased mortality risk compared with those with both undetectable cardiac troponin I and lower BNP. Integrating this multimarker approach into the routine assessment of heart failure patients will allow clinicians to more accurately identify high-risk patients who may derive increased benefit from intensive management strategies. PMID- 14564226 TI - Neurohormonal regulation and the overlapping pathology between heart failure and acute coronary syndromes. AB - An understanding of the dynamic relationship between the coronary artery and left ventricular (LV) function is important in diagnosing and treating acute coronary disease. Measurement of B-type natriuretic peptide (BNP) provides rapid and accurate identification of patients with impaired LV function, which has proven valuable in differentiating between congestive heart failure (CHF) and symptoms attributable to pulmonary etiologies. Coronary artery and ventricular pathophysiology both are characterized by injury, functional aberrations, and subsequent remodeling. Ischemia occurs in both and accounts for virtually all significant adverse outcomes. The difference in BNP elevations seen in acute ischemia compared with those observed in chronic CHF is striking: Although even small BNP elevations in acute coronary syndromes have powerful prognostic value, it is not likely that they can be effectively used as a diagnostic marker for ischemia. PMID- 14564227 TI - Combining natriuretic peptides and necrosis markers in the assessment of acute coronary syndromes. AB - Management of patients with acute coronary syndromes (ACS) is becoming more complex as the array of treatment options available to patients and physicians continues to expand. Cardiac biomarkers play an important role in risk stratification in ACS, and results of cardiac biomarker tests can be used to help guide choices between alternative therapies. In addition to biomarkers of myocyte necrosis, markers of neurohormonal activation, such as B-type natriuretic peptide (BNP), provide important prognostic information in ACS. In the future, multimarker strategies that incorporate panels of cardiac biomarkers are likely to be used for risk stratification and for pathophysiology-guided treatment in patients with ACS. PMID- 14564228 TI - Evaluation of chest pain and heart failure in the emergency department: impact of multimarker strategies and B-type natriuretic peptide. AB - In the emergency setting, acute chest pain and shortness of breath represent common patient presentations. Cardiac biomarkers including myoglobin, creatine kinase (CK)-MB, troponin, and b-type natriuretic peptide provide diagnostic and prognostic information for patients with chest pain and shortness of breath. This article reviews the use of cardiac biomarkers in the emergency department to evaluate acute coronary syndrome and congestive heart failure. PMID- 14564229 TI - New concepts in post-infarction ventricular remodeling. AB - An understanding of the process of left ventricular (LV) remodeling has led to greater knowledge of the pathophysiology of heart failure syndrome. This article examines the relationship between LV remodeling and clinical outcomes of heart failure syndrome from several different perspectives. The studies cited suggest that the post-myocardial infarction process is related to and associated with long-term progression of LV dysfunction, heart failure symptoms, and mortality. It is also demonstrated that drug therapies that slow or reverse the remodeling process appear to have favorable natural history effects in the short term as well as during long-term therapy. PMID- 14564230 TI - Reappraisal of beta-blocker therapy in the acute and chronic post-myocardial infarction period. AB - In patients presenting with acute myocardial infarction (MI), the early use of intravenous beta-blockade followed by short-term oral administration in the absence of reperfusion therapy has shown a modest reduction in mortality. In contrast, major reductions in mortality and reinfarction have been shown when beta-blockers have been used soon after an acute MI and continued long-term. These benefits were observed in trials conducted in the 1970s and 1980s, prior to the widespread use of reperfusion therapies, antiplatelet agents, and angiotensin converting enzyme inhibitors; those trials excluded patients with postischemic heart failure. Recently, the CAPRICORN trial has shown a significant reduction in all-cause mortality and reinfarction in post-MI patients with systolic dysfunction, in response to carvedilol. In spite of compelling evidence supporting the use of beta-blockers in the post-MI setting, data published by the National Cooperative Cardiovascular Project have shown that fewer than half of all post-MI patients receive beta-blockers as long-term therapy. It appears that post-MI patients with perceived contraindications, such as advanced age, diabetes, heart failure, peripheral vascular disease, and/or chronic pulmonary obstructive disease, may derive a substantial benefit from the use of beta blockers. Given the considerable evidence from randomized clinical trials, the use of beta-blockers is recommended in all post-MI patients without a contraindication, particularly in those with left ventricular systolic dysfunction. PMID- 14564231 TI - New evidence from the CAPRICORN Trial: the role of carvedilol in high-risk, post myocardial infarction patients. AB - The CAPRICORN (Carvedilol Post-Infarct Survival Control in Left Ventricular Dysfunction) trial established that the beta-blocker carvedilol reduces the risk of death in patients with left ventricular dysfunction post myocardial infarction, whether or not the infarct is complicated by clinical heart failure. Thus, the utility of the beta-blocker carvedilol is confirmed in the modern era as an adjunct to revascularization, angiotensin-converting enzyme inhibitors, aspirin, and statins. In addition, the results prompt us to review the prior studies of beta-blockers postinfarction. Critical review of CAPRICORN and earlier beta-blocker studies suggests that specific beta-blockers should be matched to specific clinical scenarios. The COMET (Carvedilol or Metoprolol European Trial) study reinforces this view by establishing that beta-blockers are not simply interchangeable agents. PMID- 14564232 TI - Diabetes, hypertension, and renal insufficiency in post-myocardial infarction cardiovascular risk. AB - The prognosis for patients who suffer myocardial infarctions (MIs) is poor, with 22% of male and 46% of female survivors being disabled by heart failure within 6 years. Many well-established risk factors for increased morbidity and mortality post-MI are closely linked to the metabolic syndrome and associated with over activation of the renin-angiotensin-aldosterone and sympathetic nervous systems. Results from numerous large-scale clinical endpoint trials have shown that blocking the deleterious effects of these systems with either an angiotensin converting enzyme inhibitor or a beta-adrenoceptor antagonist significantly reduces the risk of mortality and cardiovascular events in post-MI patients. Results from 1 recent study of the beta-blocker, carvedilol, have shown further that these benefits extend to high-risk patients with either diabetes or hypertension. PMID- 14564233 TI - The role of in-hospital initiation of cardiovascular protective therapies to improve treatment rates and clinical outcomes. AB - Patients with acute myocardial infarction (MI) face a high risk of recurrent cardiovascular events, repeat hospitalizations, heart failure, and mortality. There is compelling scientific evidence that antiplatelet therapy, beta-blockers, angiotensin-converting enzyme inhibitors, and lipid-lowering therapy reduce these risks in patients with acute MI. Despite this evidence and national guidelines, a number of studies in a variety of clinical settings have documented that a significant proportion of patients with acute MI is not being treated with these guideline-recommended, evidence-based therapies when receiving conventional care. The demonstration that initiation of cardiovascular protective medications, including lipid-lowering therapy, prior to hospital discharge for atherosclerotic cardiovascular events results in a marked increase in treatment rates, improved long-term patient compliance, and better clinical outcomes has led to the revision of national guidelines to endorse this approach as the standard of care. Physicians have been reluctant to initiate beta-blockers in post-MI patients with significant left ventricular dysfunction and/or heart failure symptoms, and this reluctance has contributed to the treatment gap. Recent studies suggest that when the beta-blocker carvedilol is initiated in acute-MI patients with left ventricular dysfunction with or without symptoms of heart failure prior to hospital discharge, it is safe and effective and improves clinical outcomes. Adopting in-hospital initiation of cardiovascular protective medications as the standard of care for patients hospitalized with acute MI could dramatically improve treatment rates and thus substantially reduce the risk of future cardiovascular events and hospitalizations and prolong life in the large number of patients hospitalized each year. PMID- 14564234 TI - Critical pathways for acute myocardial infarction. AB - Critical pathways are standardized protocols for optimizing and streamlining patient care. They are important in the management of patients with acute myocardial infarction, many of whom do not receive evidence-based therapies. Several studies have demonstrated that the development and implementation of critical pathways that follow published guidelines and are tailored to the needs and resources of each institution result in increased use of evidence-based therapies, such as aspirin and beta-blockers, and is associated with decreased mortality. Use of these protocols has also been shown to reduce unnecessary therapy and under- or overutilization of certain procedures, thereby resulting in more cost-effective treatment. PMID- 14564235 TI - SIAARTI recommendations on chronic cancer pain: a challenge for the discipline. PMID- 14564236 TI - Cardiac preconditioning and reperfusion stunning in human left ventricle. Biomolecular and clinical remarks. PMID- 14564237 TI - Effects of remifentanil on human heart electrical system. A transesophageal pacing electrophysiological study. AB - AIM: Previous studies have shown that the administration of remifentanil (a micro agonist opioid) is often accompanied by bradyarrhythmias preventable or manageable by parasympatholytic drugs. The aim of this paper is to evaluate if these negative chronotropic effects are exclusively due to an increased parasympathetic activity or to a direct action of remifentanil on heart conduction fibres. METHODS: A transesophageal pacing electrophysiological study on 40 healthy subjects scheduled for orthopaedic surgical treatment under general anaesthesia has been carried out. We determined either the correct sinus recovery time or the occurrence of Wencke-bach atrio-ventricular block in the awake state and, again, during remifentanil administration. RESULTS: In all patients either a significant depression of sino-atrial automatism or a decrease of atrio ventricular node conduction reserve was noticed. In 2 cases, in particular, a sinus arrest and a junctional rhythm, respectively, both spontaneously recovered were observed. CONCLUSION. Atropine normalized all parameters, confirming that remifentanil-associated hypokinetic cardiac phenomena are exclusively vagally mediated. PMID- 14564238 TI - Perioperative management for laparoscopic kidney donation. AB - AIM: To report a single centre's experience in the perioperative management of live kidney laparoscopic donations. METHODS. DESIGN: comparative analysis of all laparoscopic kidney donations performed between April 2000 and August 2002 and a corresponding number of "traditional surgery" donors from a historical series. SETTING: kidney transplant centre of a teaching hospital. INTERVENTIONS: 39 and 27 subjects undergoing respectively laparoscopic and traditional "open" kidney donation were studied. A standard balanced anesthesiological technique was used in both groups but, to counterbalance the reported abdominal insufflation-related kidney dysfunction, laparoscopic donors were administered an extra intravascular volume loading with colloids and crystalloids starting from the night before surgery. RESULTS: Laparoscopic donors underwent a longer procedure with a lower estimated blood loss (p=0.0001) and were intraoperatively administered with a higher amount of intravenous fluids (p<0.01); they showed less postoperative analgesic requirement (p<0.0001), a shorter ICU stay and overall hospitalisation (p<0.001), a quicker resumption of solid oral intakes (p<0.01) and full return to work (p<0.001) with no difference in the rate of postoperative complications. Diuresis resumed intraoperatively in all recipients and early graft function did not differ in the 2 groups although serum creatinine declined earlier, but not significantly, in those receiving kidneys by the traditional method. CONCLUSION: Kidney laparoscopic donation does not require a particularly complex or expensive anaesthetic management or approach; it is advisable to adopt strategies to counterbalance laparoscopy-associated abdominal hypertension. PMID- 14564239 TI - Replacement treatment with protein C in an 18-year-old man with meningococcal sepsis and purpura fulminans. AB - In separate studies on Neisseria meningitidis sepsis, Powars and Fiynvandraat suggested that low protein C levels may be responsible for disseminated intravascular coagulopathy and purpura fulminans. Following on this observation, we used protein C concentrate in an 18-year-old male patient with septic shock and purpura fulminans. The patient's coagulation parameters were seriously altered: AT 45%; protein C 21%; PT 50%; platelets 55000; D-dimer 2400. Early treatment included immediate administration of 3000 IU of antithrombin and intensive therapy: antibiotic therapy, volemic replacement, supported by inotropic drugs and oxygen therapy. Given the patient's low protein C levels and the progression of purpura, replacement therapy with protein C concentrate was instituted. The initial dose of 80 IU/kg/bw (5600 IU) in bolus, was adjusted according to blood laboratory values and then set at 2000 IU every 8 hours for 4 days. An increase in protein C was observed (78%) after the 1st administration, while the D-dimer levels fell by 50%. By day 7, the patient's cardiocirculatory conditions had stabilized and the coagulation parameters had normalized; the patient was discharged from the intensive care unit. Protein C replacement therapy normalized the coagulation parameters and blocked the evolution of the skin injuries. PMID- 14564240 TI - SIAARTI recommendations on the assessment and treatment of chronic cancer pain. PMID- 14564241 TI - What status does day surgery hold in Italy? What opportunities lie ahead for anesthesiologists? PMID- 14564243 TI - Original insight into continuous cardiac output monitoring: "TruCCOMS". Correlation with other methods. AB - AIM: Of all technical devices used for continuous or intermittent monitoring of cardiac output, in our clinical practice during the last year, we tested a new system, the true continuoas cardiac output monitoring system (TruCCOMS), for the continuous real time measurement of cardiac output. The purpose of this study was to compare the accuracy, reliability and promptness of TruCCOMS with other systems and methods of cardiac output (CO) measurement such as pulsion continuous cardiac output (PiCCO) and end diastolic Area (EDA) determination by trans esophageal-echocar-diography (TEE), keeping as gold standard for CO measurement the thermodilution method by Swan-Ganz. METHODS: Sixteen male patients, aged 50 to 60 years, with ejection fraction (FE) >50%, EUROSCORE=1, who underwent CABG surgery with circulation extra corporeal (CEC), were analysed with all methods mentioned above. The measurements were obtained at different phases: pre-CEC; post-CEC; and Intensive Care Unit (ICU). In the ICU setting, 5 patients classified as under-filled with a pulmonary capillary wedge pressure (PCWP) =/<8 mmHg were filled until a PCWP gs;13 mmHg in order to evaluate the promptness of the various systems to detect the new condition. RESULTS: The statistical analysis of data obtained in our survey, clearly demonstrates that TruCCOMS is at least equivalent to average thermo-dilution cardiac output (AvTDCO), with the advantage of being continuous, real time and, furthermore, with no need for physician intervention/interference. CONCLUSION: In our experience the TruCCOMS seems to be an ideal method for continuous cardiac output (CCO) monitoring compared with the other bedside systems challenged. PMID- 14564242 TI - Atracurium, cisatracurium, vecuronium and rocuronium in patients with renal failure. AB - AIM: The cumulative index, the recovery, the onset and the duration of action, of atracurium, cisatracurium, vecuronium and rocuronium in uremic patients undergoing kidney transplantation compared to healthy patients undergoing general surgery were studied. METHODS: In all patients (64 uremic vs 62 "healthy" patients) after anesthesia induction, atracurium 0.5 mgxkg(-1) or cisatracurium 0.15 mgxkg(-1) or vecuronium 0.1 mgxkg(-1) or rocuronium 0.6 mgxkg(-1) were administered, and at the end of surgery when T1 reached 25% neostigmine 0.05 mgxkg(-1) was given. Neuro-muscu-lar transmission was monitored by accelerometry (TOF-GUARD, Organon). RESULTS: Cumulative index of vecuronium (1.3+/-0.1 vs 1.06+/-0.11, p<0.001) and rocuronium (1.45+/-0.18 vs 1.04+/-0.16, p<0.001), recovery index (time of T1 25-75) of atracurium (14.2+/-5 vs 9+/-4, p<0.005), cisatracurium (18.7+/-3 vs 9.1, p<0.001), vecuronium (18.5+/-3 vs 12.5+/-3, p<0.001) and rocuronium (18+/-6 vs 11+/-4, p<0.001) and interval T1 25% to TOF 0.8 of cisatracurium (20.5+/-1.2 vs 16+/-2.1, p<0.001) and vecuronium (27+/-6.3 vs 20+/-3.3, p<0.001) were longer in uremic patients. The onset time and the duration of action of atracurium, cisatracurium, vecuronium and rocuronium were similar in all groups compared to controls one. CONCLUSION: In patients with renal failure the use of atracurium, cisatracurium, vecuronium and rocuronium is suitable and predictable in terms of onset, and duration of action. Care has to be taken to vecuronium and rocuronium cumulative index. Neuromuscular trasmission has to be always monitored. PMID- 14564245 TI - Anaestethic problems in Sanfilippo syndrome. A rare case of adult patient. AB - The authors report the case of a female patient (41 years old) affected by mucopolysaccharidosis type III or Sanfilippo syndrome submitted to a gynecologic surgical procedure and describe the main anesthesiologic problems. A sub arachnoid anesthesia with hyperbaric Bupivacain 0.5% was used. This technique proved to be safe and convenient without peri- and postoperative complications. PMID- 14564244 TI - Demand and availability of Intensive Care beds. A study based on the data collected at the SUEM 118 Central of Padua from October 1996 to December 2001. AB - AIM: This study aims to evaluate the management of intensive care beds according to the demands received by the SUEM 118 of Padua. It has been carried out by examining the reports drawn up by SUEM physicians from October 1996 to December 2001. The study rated the number of patients for whom an admission to the Intensive Care Unit (ICU) was required, according to the specific clinical situation at the moment of the request. A secondary objective was to evaluate if the critically ill patients had been admitted and treated in the most appropriate medical facility. METHODS: The research is based on 7 087 reports concerning a population of adult and pediatric patients for whom an ICU bed was required in the period previously mentioned. For each report, it analyses the following data (keeping them anonymous): date of demand, main pathology and severity of clinical condition, sex and age, provenence and destination. RESULTS: Even though the number of annual demands for an ICU bed made to SUEM Central 118 has remained unchanged (approximately 1 350 per year), the number of beds made available in the operating rooms of the Hospital of Padua markedly increased. What has been experienced so far, and the data collected in this study has revealed, was that the requests for an intensive treatment for the overall population (hospitalized and non hospitalized) increased disproportionally in relation to the availability of ICU beds. In fact, the total number of hospitalizations in the different ICUs rose steadily year by year (from 3 495 in 1996 to 4 640 in 2001). CONCLUSION: The Hospital of Padua is a landmark center for patients who need specialized treatment. It is therefore important to increase the assistance and safety standards of its ICUs. In recent years there has been a great need for specialized ICUs either for more aggressive procedures (neurosurgical, cardiosurgical, respiratory, cardiologic, etc.) or for the increased use of adequate and invasive treatment for advanced diseases. The available resources of ICU beds should be more rationally distributed between the peripheral and the Regional Hospitals, since the activation of an ICU bed in the operating theatre is a valid, transient option. PMID- 14564247 TI - Analgesic therapy services and levels of essential care. PMID- 14564246 TI - Intensive therapies and new methods of dying. PMID- 14564248 TI - Inhalation anaesthesia and myocardial preconditioning. PMID- 14564249 TI - Thrombelastography. Present and future perspectives in clinical practice. AB - RESULTS: Thrombelastography (TEG) is a method for evaluating the viscoelastic properties of the blood clot, from its formation to its lysis. All major surgeries may be associated with massive blood loss, with blood component transfusion therapy often becoming mandatory. The clinician's goal is thus to optimize and possibly minimize blood components usage. To this end, TEG allows for a qualitative and dynamic analysis of the specific blood clotting process, from clot formation through its lysis, highlighting alterations at every single step in the cascade. With TEG is thus possible to know if bleeding is due to a failure to provide adequate surgical hemostasis, if there is platelet dysfunction, or to detect anomalies in coagulation proteases or their inhibitors, or if the blood loss is associated to early, excessive fibrinolysis. The theoretical advantages of TEG are the ease of performing the test, the fast sample reading times (now 30 minutes) and the informative PMID- 14564250 TI - A prospective, observational evaluation of a new supraglottic airway: the PAXpress. AB - AIM: Little information is available on efficacy and safety of the new supraglottic device, PA(Xpress), in anesthetized patients. The aim of this prospective observational study was to evaluate the use of this new supraglottic device in clinical practice. METHODS: Ninety-one patients (mean age: 45+/-11 years) undergoing extremity or minor peri-phery procedures (mean duration 53+/-18 min), with a supraglottic airway were prospectively studied in 10 hospitals, using a simple questionnaire with data concerning anthropometric variables, surgical procedure, and occurrence of untoward events during PA(Xpress) placement, anesthesia maintenance, or postoperative period. RESULTS: First attempt placement was achieved in 74 patients (82.2%), while 9 patients (10%) required 2 attempts, and 7 patients (7.7%) required more than 2 attempts. In only 2 patients (2%) airway control was not achieved with the studied device. The mean time required to achieve successful placement was 49+/-18 s (range 4-300 s). Mechanical ventilation was effectively maintained in all studied patients, and no severe adverse events were reported during the procedure. Blood on the device was observed in 54% of cases, and this was associated with an incidence of sore throat of 26% in the recovery area and 13% after 6 hours from the end of surgery. CONCLUSION: Although further comparative, randomized studies should be advocated to better evaluate the use of this new supraglottic device, this prospective, observational study demonstrated that PA(Xpress) provides safe and effective airway control during mechanical ventilation in up to 98% of cases. PMID- 14564251 TI - Regional filter heparinization for continuous veno-venous hemofiltration in liver transplant recipients. AB - AIM: To study safety, clinical and operative feasibility of continuous veno venous hemofiltration (CVVH) with anticoagulation only of the filter in patients at risk for bleeding. METHODS: This prospective, comparative, non randomised study was completed at an intensive care unit of a teaching NHS hospital. Sixteen liver transplant (LT) recipients with acute renal failure needing CVVH were treated with a regional anticoagulation protocol (heparin and protamine were administered respectively pre- and post-filter) and compared to 11 critically ill subjects who received a standard low-heparin treatment. Activated coagulation time (ACT) monitoring was used to adjust anticoagulation and heparin neutralization. RESULTS: Mean circuit life was 35.8+/-13.6 hours (95% CI 28.5 43.1) in patients receiving regional anticoagulation and 34.4+/-14 hours in controls (95% CI 25.5-43.3; p=0.7). Fourty-eight circuits (47.5% of the total) in the heparin-protamine group had a life-span longer than 30 hours and other 22 (21.7%) were changed intentionally after 24 hours of use in absence of clots. None of the patients in both the studied groups had bleeding or hemodynamic complications and their azotemic control was always satisfactory. CONCLUSION: In LT recipients, regional anticoagulation can achieve a circuits life-span comparable to that from systemic anticoagulation with satisfactory safety and simplicity of use. PMID- 14564252 TI - Sepsis: state of the art. AB - In recent years, we have considerably widened our knowledge of the pathophysiology of sepsis and some procedures, aiming both to relieve symptoms and control the inflammation/coagulation reaction, have proven to be effective in increasing survival. This improves when mechanical ventilation is applied with low tidal volumes, fluid replacement and the use of cardioactive drugs are titrated on the oxygen saturation of hemoglobin in the central venous system and blood glucose does not exceed certain limits. It is also evident that inflammation and coagulation are closely related to each other. The inhibition of only one pathway, such as the inhibition of inflammation with high dosage steroids or the inhibition of coagulation with antithrombin, does not produce a survival improvement. The only molecule which has proven to be notably effective in reducing mortality is Activated Protein C interacting on coagulation/fibrinolysis, as well as on inflammation processes. Multinodal modulation of several interdependent processes may be the probable reason for the proven effectiveness of this treatment. PMID- 14564253 TI - Postoperative analgesia after preincisional administration of remifentanil. AB - AIM: The aim of this study was to assess postoperative analgesia after preincisional and postincisional administration of remifentanil. METHODS: Randomized trial, 24 hours. SETTING: University hospital, hospitalized care. PATIENTS: 48 adult patients scheduled for lumbar vertebral surgery. INTERVENTIONS: in group R5, patients received an infusion of 0.2 microg kg(-1) min(-1) remifentanil over 5 minutes, followed by a break of 15 minutes before anesthesia was started. Anesthesia was induced by infusion of 0.25 microg kg(-1) min(-1) remifentanil and a bolus of 1.5 microg kg(-1) propofol, followed by a continuous infusion of 2 to 3 microg kg(-1) h-1 propofol and 0.25 microg kg(-1) min(-1) remifentanil until end of anesthesia. In group R20, patients received 0.05 microg kg(-1) min(-1) remifentanil over 20 minutes before the induction of anesthesia. In group RL, anesthesia was induced and maintained with propofol. After surgery began, a remifentanil infusion of 0.5 microg kg(-1) min(-1) was given for 50 minutes, then reduced to 0.25 microg kg(-1) min(-1). The total remifentanil doses were similar in the 3 groups. MEASURES: patients used patient controlled analgesia (piritramide) for postoperative pain management. They recorded pain on a numeric rating scale every half hour. STATISTICS: Kruskal Wallis test, pairwise Mann-Withney U-test, orthogonal polynomials (pain scores). RESULTS: PATIENTS given postincisional remifentanil (RL) had the slowest decrease in postoperative pain scores (p<0.01) and the highest cumulative piritramide consumption (p<0.08). CONCLUSION: The preincisional administration of remifentanil followed by a continuous infusion of 0.25 microg kg(-1) min(-1) appears to reduce pain scores and piritramid consumption when compared with a postincisional regimen. PMID- 14564254 TI - Alemanno's brachial plexus block ten years later: topographic study of the anesthetized areas. AB - AIM: The study analyzes the extension of anesthesia induced by Alemanno's brachial plexus block technique on the various areas of competence of the different nerves. METHODS: The study was conducted on 58 patients in ASA classes 1 and 2 scheduled to receive shoulder arthroscopy. At the end of the operation, about 2 hours after induction of anesthesia, extension of anesthesia was evaluated by the pin-prick test. RESULTS: Anesthesia was achieved in 100% of cases for the circumflexus, musculocutaneous and radial nerves; the median nerve escaped in 7% of cases, the medial cutaneous nerve of forearm in 20%, the ulnar nerve in 27.5%, the medial cutaneous nerve of arm and intercostobrachial nerves in 46%; no major complications were associated with the technique. CONCLUSION: Alemanno's technique is recommended for operations on the shoulder and humerus, whereas more peripheral techniques may be preferable for interventions on the forearm and hand. PMID- 14564255 TI - Infective endocarditis due to Staphylococcus aureus involving three cardiac valves. A case study. AB - A 20-year-old woman, diagnosed with coarctation of the aorta, situs viscerum inversus, and bicuspid aortic valve, underwent corrective surgery for the coarctation. After a postoperative neurological state that suggested a spinal lesion, corticosteroid therapy was initiated and the patient was discharged early from the unit to begin a motor rehabilitation program. Following the dehiscence of the thoracotomy surgical wound, a severe infective clinical picture, sustained by methicillin-resistant S. Aureus (MRSA), became evident with a diagnosis of bacterial endocarditis involving the aortic, mitral and tricuspid valves and caused the patient's death due to septic shock complicated by ARDS. According to the authors, the early discharge of the patients after such a complex operation, the eccessive lengthening of the steroid therapy that would have contribuited to delay the diagnosis, causing the lack of preventing identification of the first signs of infection and the impossibility for the patient to have another operation (involving 3 valves) are conclusive elements that led to the above mentioned complications. PMID- 14564256 TI - [Rhabdomyolysis associated with respiratory infection in chronic psychiatric patients during neuroleptic treatment]. AB - Rhabdomyolysis is a disorder characterized by skeletal muscle injury and fatal complications at times. The causes of rhabdomyolysis are usually traumatic and non-traumatic, such as neuroleptic malignant syndrome and rhabdomyolysis associated to septicemia. The cases of 2 schizophrenic patients with rhabdomyolisis during pneumonia infection and neuroleptic therapy are reported. At admission, both patients had important respiratory distress and hyperthermia; the clinical conditions required endotracheal intubation. Blood and urine cultures were always negative, while the bronchial sputum culture was positive. The diagnosis of rhabdomyolysis was confirmed by myoglobinemia dosage and ortholuidine test. Pneumonia infection was treated with antibiotic specific therapy whereas renal failure was treated with adequate hydratation and strained diuresis. The absence of muscle rigidity, the improvement of X-r images and the reduction of corporeal temperature, during antibiotic treatment, excluded neuroleptic malignant syndrome. The impro-vement allowed extubation and discharge of the patients from intensive care unit. In both cases neuroleptic malignant syndrome was excluded, therefore rhabdomyolysis was the consequence of pneumonia infection or of a combination of factors capable to cause an important damage of skeletal muscles. PMID- 14564257 TI - Airway management in emergency medical services. PMID- 14564258 TI - A new method of immobilization after traumatic anterior dislocation of the shoulder: a preliminary study. AB - This preliminary prospective study was conducted to determine whether immobilization with the arm in external rotation would decrease the rate of recurrence after initial traumatic anterior dislocation of the shoulder. Forty patients with initial shoulder dislocations were assigned to (1) conventional immobilization in internal rotation (IR group, n = 20) or (2) a new method of immobilization in external rotation (ER group, n = 20). The recurrence rate was 30% in the IR group and 0% in the ER group at a mean 15.5 months. The difference in recurrence rate was even greater among those who were aged less than 30 years (45% in the IR group and 0% in the ER group). Immobilization with the arm in external rotation is effective in reducing the rate of recurrence after initial dislocation of the shoulder. PMID- 14564259 TI - Rotator cuff repair in patients with type I diabetes mellitus. AB - Insulin-dependent diabetes mellitus is associated with shoulder stiffness and a propensity toward postoperative wound complications and infection. We compared our results of open repair of full-thickness rotator cuff tears in 30 diabetic patients with those of a matched, nondiabetic population. No differences were observed in preoperative range of motion, although at a mean of 34 months, significant differences in shoulder active range of motion and passive range of motion were found postoperatively at 6 weeks, 6 months, and final follow-up (P <.05). On the basis of American Shoulder and Elbow Surgeons shoulder scoring, there were 27 (90%) and 28 (93%) good or excellent results in the diabetic and comparison groups, respectively. Complications occurred in 5 diabetic patients (17%), with 2 failures (7%) and 3 infections (10%), as compared with 1 failure (3%) and no infections in the comparison group. Repair of the diabetic rotator cuff may be performed with the expectation of improved motion and function, although less than nondiabetic counterparts. The surgeon should remain cognizant that a higher rate of complications, infection in particular, may occur after rotator cuff repair in the diabetic population. PMID- 14564260 TI - Muscle biopsy investigations on neuromuscular insufficiency of the rotator cuff: a contribution to the functional impingement of the shoulder joint. AB - The possibility that functional impingement results from muscular imbalance has been discussed. This study investigates whether disturbance of muscular coordination is reflected in a shift in the balance of fast-twitch and slow twitch fibers, as fast-twitch fibers enable a rapid reaction or contraction and slow-twitch fibers enable slow contractions and sustained performance. The supraspinatus and deltoid muscles in 37 patients with a supraspinatus syndrome and partial or complete rupture of the rotator cuff underwent biopsy. All three groups of patients showed a reduction in size, increased variability, and change in the frequency distribution of fiber cross sections. The fast-twitch fibers were especially affected. These findings were confirmed objectively by morphometric measurements. It might be concluded from the changes mentioned above that disturbance in coordination of the musculature can cause a functional impingement. We term this neuromuscular insufficiency of the rotator cuff. The mechanical factors relating to subacromial impingement may only have a role in promoting its manifestation by restricting the compensatory range of the joint. PMID- 14564261 TI - The belly-press test for the physical examination of the subscapularis muscle: electromyographic validation and comparison to the lift-off test. AB - The purpose of this study was to determine the validity of the belly-press test as a clinical test for the subscapularis muscle with the use of electromyography (EMG). In addition, the belly-press and lift-off tests were compared to determine whether the two physical examination techniques are equivalent in their evaluation of the upper and lower portions of the subscapularis muscle. EMG data of 7 muscles (upper subscapularis, lower subscapularis, infraspinatus, latissimus dorsi, teres major, pectoralis major, and supraspinatus) were studied in 16 healthy volunteers. Average EMG amplitudes were contrasted within and between tests. Both the belly-press and lift-off tests activated the upper and lower portions of the subscapularis muscle greater than all other muscles, indicating that both tests are valid and specific for evaluation of the subscapularis muscle (P <.05). The belly-press test was found to activate the upper subscapularis muscle significantly more than the lift-off test (P <.05), whereas the lift-off test was found to pose a significantly greater challenge to the lower subscapularis muscle than the belly-press test (P <.05). These findings may improve the clinical testing and assessment of the subscapularis muscle. PMID- 14564262 TI - Impaction grafting improves the fit of uncemented humeral arthroplasty. AB - This study tested the hypothesis that the excellence of the fit between a humeral prosthesis and the humerus can be optimized by impaction grafting with cancellous bone. Ten paired human humeri were prepared for insertion of a humeral implant by a standard surgical technique. One humerus from each pair was randomized to receive a 10-mm humeral component with cancellous impaction autografting, whereas the other was inserted without grafting. After insertion of a polymethylmethacrylate model of the prosthesis, computed tomography scans were obtained with 3-mm sections and the void areas of each section measured by use of NIH Image. The data demonstrate that cancellous impaction grafting significantly reduces the void between the prosthesis and the humerus (P <.001). The effect was most marked in the proximal and middle thirds of the implant. We conclude that cancellous impaction grafting is an effective method for improving the quality of fit between the humerus and the proximal two thirds of the humeral prosthesis. PMID- 14564264 TI - Stabilization of acute, complete acromioclavicular joint dislocations with a new C hook implant. AB - Although coracoclavicular fixation is currently popular for type III acromioclavicular (AC) dislocations, a surgical gold standard is lacking. The purpose of this study was to evaluate the preliminary outcome of surgical treatment of complete AC dislocations with a new nitinol C hook implant. When the implant is cooled (<5 degrees C), it softens enough to be easily inserted under the coracoid process with a hole drilled in the clavicle. Cooling is induced with ice water. When the implant reaches body temperature, it hardens and anatomic reduction is achieved. Patients with acute type III AC dislocations were prospectively evaluated. The AC ligament was reinserted with the use of a bone anchor, and the position of the joint was restored by fixing it with a new C hook. Clinical and radiographic control checkups were carried out at 3, 8, and 12 weeks and 2 years postoperatively. Certain patient-related variables, functional status, symptom severity, and patient satisfaction were assessed. By 12 weeks, all patients had achieved full functional status. Radiographs showed accurate anatomic reduction. Overall subjective satisfaction was very good in all cases. No complications or implant failures occurred. On the basis of this pilot study, the new C hook implant provides secure anatomic reduction with very good functional recovery and patient satisfaction. The main benefit of the implant is the ease of insertion. It preserves the articular surfaces and allows slight movement of the AC joint during abduction of the arm. The C hook implant is a new surgical concept with potentially better patient recovery. PMID- 14564263 TI - Treatment of post-traumatic humeral diaphyseal nonunion with bone loss. AB - Four patients with post-traumatic nonunion and shortening of the humeral diaphysis were treated with a hybrid advanced Ilizarov technique. The mean age of the patients was 32 years, and the mean total amount of humeral shortening was 6.63 cm. Three nonunions were atrophic and infected, and one was hypertrophic. All patients obtained union of the humeral fracture with resolution of infection at a mean external fixation time of 8 months. Restoration of normal humeral length was achieved in two patients, with a third having a residual discrepancy of 1 cm. The final patient, who had an infected nonunion with 11 cm of total humeral shortening, had a residual limb length discrepancy of 3 cm. All had improvement in shoulder and elbow motion after treatment. Superficial pin tract infections were seen in all patients, but all responded to pin-site care and oral antibiotics. Two patients had three refractures after removal of the fixator, two of which were treated by a second application of an Ilizarov frame and one by a cast. All patients had reduced pain and improved function at completion of the treatment. The Ilizarov method, though not a panacea for all humeral nonunions with extensive bone loss, does offer a viable salvage procedure in this unusual and often complex clinical problem. PMID- 14564265 TI - Chronic shoulder dislocations. AB - Thirty-two patients with chronic dislocation of the shoulder were diagnosed over a 5-year period at King Edward VIII Hospital, Durban, South Africa. Thirty-one patients had an anterior subcoracoid dislocation. One patient had a chronic posterior dislocation. The duration of dislocation ranged from 6 days to 2 years. Six patients were epileptic, and it was an affliction of all ages. None of the patients had a vascular deficit, but 4 had neurologic damage on presentation. Closed manipulation was only successful in 1 patient. Ten patients' shoulders were left unreduced in the dislocated position. Twenty-one patients, including the patient with a single posterior dislocation, underwent open reduction. The author treated 10 patients with chronic anterior dislocations surgically. The coracoid was transferred to the glenoid, and acromiohumeral K-wire fixation was used for 4 weeks in this group. These 10 cases, as well as 5 cases left unreduced, were followed up for more than 2 years. The results were evaluated by the system proposed by Rowe and Zarins and showed that surgically treated patients fared better than those whose shoulders were left unreduced regardless of the duration of the dislocation or the age of the patient. This series differs from previous reports in several respects. All but one of the dislocations were anterior, the incidence of epilepsy was lower, and successful reduction by manipulation was rarely achieved. In all 10 cases in which the author performed the operation, the shoulder was successfully relocated without neurovascular complications. Dissection of the axillary vessels and nerves was never necessary. PMID- 14564266 TI - Shoulder girdle neoplasms mimicking frozen shoulder syndrome. AB - Shoulder pain and immobility comprise a multifactorial disorder apparently affected by pain inhibiting joint motion. As the syndrome is very common, many patients do not undergo detailed imaging studies before treatment. This study compared a series of 7 patients in whom a neoplasm was the underlying cause for the stiff shoulder with a series of 50 patients with primary or secondary frozen shoulder. In addition to a detailed history being taken, the Disabilities of the Arm, Shoulder, and Hand (DASH) upper limb outcomes data collection questionnaire was completed and physical examination, radiography, ultrasonography, and bone scanning were performed in all cases. In the cases of tumor, the presenting symptom was a stiff shoulder without radiographic abnormality in 7 of 67 patients with shoulder girdle neoplasms who were seen at our musculoskeletal oncology clinic. The tumors included osteoid osteoma, osteoblastoma, metastatic carcinoma, chondrosarcoma, periosteal lipoma, and acute lymphoblastic lymphoma. The diagnosis was established in all cases by an area of focal isotope uptake demonstrated by a routine technetium 99 methylene diphosphonate bone scan. In a single case of metastatic colon carcinoma, the diagnosis could only be established by magnetic resonance imaging, as the radiographs were normal and the bone scan demonstrated diffuse uptake over the proximal humerus. The patients whose frozen shoulder was caused by an underlying tumor were significantly younger and had a lower fatigue/energy dimension score on the RAND Short Form-36 health survey. The most useful diagnostic test appears to be a discrete area of bony tenderness, present in 7 of 7 patients with tumor and in only 5 of 50 patients in the control group. Although an underlying tumor is a rare cause of frozen shoulder syndrome, the potential grave consequences of misdiagnosis and the possibility of performing an unnecessary and ineffective invasive procedure should prompt physicians to increased vigilance. In patients with discrete bony tenderness elicited by light tapping, a bone scan should be ordered and magnetic resonance imaging should be considered. PMID- 14564267 TI - Surgical treatment of partial distal biceps tendon ruptures through a single posterior incision. AB - The purpose of this study was to describe a novel technique for repair of partial distal biceps tendon ruptures through a single posterior incision. Eight patients with partial distal biceps tendon ruptures had the tear confirmed and repaired through a single posterior incision. All were men. The mean age was 50 years (range, 36-60 years). Postoperatively, immediate active and passive motion was instituted. Seven of eight patients returned to their previous professions. The mean postoperative American Shoulder and Elbow Surgeons elbow score was 96 (range, 89-100). Six of the patients were completely satisfied, and all were much improved. There were no complications. Surgical confirmation and repair of symptomatic partial distal biceps tendon ruptures can be performed through a single posterior incision. The morbidity of the anterior exposure can be avoided and the tendon readily explored and reattached with excellent patient outcomes. The security of repairing the tendon directly into a trough in bone and suturing over cortical bone permits immediate rehabilitation without immobilization in a cast or splint. PMID- 14564268 TI - Intra-articular fluid volume and restricted motion in the elbow. AB - Eleven fresh-frozen cadaveric upper extremities were studied to determine the effect of intra-articular fluid volume on the range of motion of the elbow. The flexion arc of the cadaveric elbows was measured before and after incremental injections of saline solution. Linear regression analysis showed that the relationship between the flexion arc and intra-articular fluid volume is linear up to a total volume of 25 mL. The flexion arc of motion decreased by a mean of 2.1 degrees per milliliter of fluid injected. A mean volume of 10.8 mL of saline solution decreased the flexion arc to the point where surgery has traditionally been recommended for type II radial head fractures. On the basis of these findings, the use of post-aspiration range of motion should be reconsidered as a criterion for surgical treatment of radial head fractures. PMID- 14564269 TI - Elbow joint stability following experimental osteoligamentous injury and reconstruction. AB - Elbow joint dislocation was simulated in cadaveric specimens to quantify laxity induced by radial head and coronoid process lesions, either alone or in combination with collateral ligament insufficiency. The effects of lateral ligament reconstruction and radial head prosthesis replacement were also considered. Absence of the radial head and the coronoid process induced rotatory laxity of 145% and 128% (both P <.01), respectively, compared with the intact joint. When both were absent, the joints subluxated regardless of collateral ligament status. Isolated radial head prosthesis implantation prevented this subluxation, and laxity almost normalized. Lateral collateral ligament reconstruction prevented major laxity even in the absence of the radial head. Lateral collateral ligament reconstruction and radial head prosthetic replacement yielded restraint against gross instability in the maximal unstable situation (terrible triad). The lateral collateral ligament is the prime stabilizer to external rotation, and reconstruction of this alone, even with an absent radial head, is beneficial. PMID- 14564270 TI - Three-dimensional corrective osteotomy for cubitus varus in adults. AB - In 23 adult patients, cubitus varus deformity was corrected by 3-dimensional osteotomy. During surgery, not only varus but internal rotation, flexion extension deformity of the elbow, and lateral protrusion of the distal fragment were simultaneously addressed. The mean age of the patients was 26 years. Three showed tardy ulnar nerve palsy. The follow-up period after osteotomy averaged 1 year 10 months. The humeral-elbow-wrist angle improved from a mean 26 degrees of varus preoperatively to a mean of 3 degrees of valgus postoperatively. The mean internal rotation angle improved from 25 degrees to 5 degrees. As there was no recurrence of the deformity, this method of 3-dimensional corrective osteotomy for the treatment of cubitus varus in skeletally mature adults is recommended. PMID- 14564271 TI - Surgical reconstruction for posterolateral rotatory instability of the elbow. AB - We reviewed the results of 10 patients (10 elbows) who underwent surgical reconstruction for clinically symptomatic posterolateral rotatory instability of the elbow. The symptoms resulted from previous dislocation or a hyperextension or varus stress injury of the elbow. Two elbows had underlying preexisting varus deformity. Surgical reconstruction was performed with a tendon graft in 6 elbows and reattachment of the lateral ulnar collateral ligament to the humerus in 4. Postoperatively, no patient had residual instability or a positive pivot shift test in the elbow. Results were graded as excellent or good in 8 and fair in 2. All patients with excellent results had surgical reconstruction with a tendon graft. Subjective assessment revealed that all 10 patients were satisfied with the outcome of the surgery. Accurate recognition of posterolateral rotatory instability of the elbow is important for appropriate management. Surgical ligament reconstruction or repair is the most favorable treatment option for restoration of normal elbow function. The choice of reconstruction with a tendon graft appeared to produce better results than the reattachment of the injured ligament by itself in achieving an excellent outcome. PMID- 14564272 TI - Reoperation rate after elbow synovectomy in rheumatoid arthritis. AB - The purpose of this study was to evaluate the reoperation rate of elbow synovectomy in patients with rheumatoid arthritis. A total of 103 synovectomies were performed in 88 patients (61 women) with rheumatoid factor-positive rheumatoid arthritis with a mean follow-up of 5.2 years (range, 1-8 years). The survival rate after elbow synovectomy (free from reoperation) was 77% (95% confidence interval, 66%-85%) at 5 years. Eight resynovectomies and fourteen total elbow replacements were performed during the follow-up. No significant improvement in range of motion was detected after synovectomy, but pain relief and patient satisfaction were favorable. Elbows were classified preoperatively (before primary synovectomy) with the Larsen system. All resynovectomies were performed for elbows of grade 0-2 destruction. A significant difference was found between early (Larsen grade 0-2) and late (Larsen grade 3) synovectomies in relation to elbow replacement (P =.002) during the follow-up. Late synovectomy yielded more temporary pain relief with a high rate of elbow arthroplasties. PMID- 14564273 TI - EndoButton-assisted repair of distal biceps tendon ruptures. AB - This report represents a multifactorial investigation of a new technique in which a titanium EndoButton was used for repair of distal biceps tendon ruptures. Cadaveric cases were used to demonstrate the anatomic efficacy and safety of the procedure. Biomechanical testing was done to compare the fixation strength of traditional techniques with the EndoButton repair. Finally, clinical results of the repair were evaluated. In 15 fresh-frozen cadavers the mean distance of the button from the posterior interosseous nerve was 9.3 mm. Instron testing showed a mean pullout strength of 253 N for the Mitek G4 Superanchor, 177 N for a conventional bone bridge, and 584 N for the titanium button. The button was 3 times stronger than the bone bridge (P =.0001) and 2 times stronger than the Mitek anchor (P =.0007). Fourteen patients who had their tendons repaired by this technique were evaluated at a mean of 20 months postoperatively. BTE (Baltimore Therapeutic Equipment, Baltimore, MD) testing revealed recovery of 97% of flexion strength and 82% of supination strength. Patients were able to participate in an aggressive rehabilitation program and were able to regain strength and function rapidly, with satisfactory return to preinjury activities and occupations. This technique is safe, simple, and stronger than any currently available anchoring techniques and gives the surgeon a choice in bone preparation. By using a single anterior elbow approach, the development of synostosis associated with two incision techniques can be minimized. PMID- 14564274 TI - Spinoglenoid septum: a new anatomic finding. AB - The spinoglenoid ligament has been implicated in compression of the suprascapular nerve in cases with isolated atrophy of the infraspinatus muscle. We dissected 32 shoulders of 16 embalmed cadavers. We could detect a spinoglenoid ligament in only 5 shoulders, and it was thin, loose, and weak. Instead of the spinoglenoid ligament, we observed a septum formed by the thickening of the fascial cover of the distal third of the supraspinatus and infraspinatus muscles. It originated from the spinoglenoid notch and extended into the posterior capsule. We named this structure the spinoglenoid septum. The suprascapular nerve passed between the bony margin of the spinoglenoid notch and the medial concave margin of the spinoglenoid septum. This septum may be a cause of dynamic compression of the suprascapular nerve. PMID- 14564275 TI - Measurement of glenoid version: conventional radiographs versus computed tomography scans. AB - Glenoid version seems to play an important role in the stability and loading of the glenohumeral joint. The purpose of this study was to compare measurements of glenoid version on axillary views and computed tomography (CT) scans. Radiographs and CT scans of 25 patients evaluated predominantly for glenohumeral joint instability and 25 patients after implantation of a total shoulder prosthesis were analyzed by 3 independent observers. In all patients glenoid version was determined on an axillary view and on a CT scan at the mid-glenoid level. The mean glenoid version measured on CT scans was 3 degrees of retroversion in the instability group (range, 7 degrees of anteversion to 16 degrees of retroversion) and 2 degrees of anteversion in the total shoulder prosthesis group (range, 16 degrees of anteversion to 23 degrees of retroversion). Glenoid retroversion was overestimated on plain radiographs in 86%. The mean difference between measurements of glenoid version on axillary views and CT cuts was 6.5 degrees (range, 0 degrees -21 degrees ), and the coefficient of correlation between these measurements was 0.33 in the instability group and 0.67 in the prosthesis group. In conclusion, glenoid version cannot be determined accurately on standard axillary radiographs, either preoperatively or postoperatively. Studies that assess the role of glenoid component orientation should use a reproducible method of assessment such as CT. PMID- 14564276 TI - An anatomic study of the effects on the suprascapular nerve due to retraction of the supraspinatus muscle after a rotator cuff tear. AB - The purpose of this anatomic study was to assess the risk to the suprascapular nerve by measuring the tension on the nerve and the angle between the nerve and its motor branch at the scapular notch with medial supraspinatus tendon retraction. Twelve shoulders in six cadavers were dissected to evaluate the branching point of the first motor branch of the suprascapular nerve, the change in angle between the nerve and its first motor branch at the scapular notch with retraction of the supraspinatus tendon, and the resulting tension on the nerve. The first motor branch originated at the notch in 9 cadavers, just proximal in 1, and just distal in 2. With the supraspinatus muscle in its anatomic position, the suprascapular nerve and its first motor branch angle measured 142.6 degrees at the scapular notch. After retraction of the supraspinatus, the angle markedly decreased to 98.7 degrees and 34.6 degrees with 1 cm and 5 cm of medial retraction, respectively. The motor branch was taut in all specimens at 2 to 3 cm of retraction. Medial retraction of the supraspinatus tendon drastically changes the course of the suprascapular nerve through the scapular notch, creating increased tension on the nerve. The degree of rotator cuff muscle atrophy frequently observed after a massive tear may be explained by increased tension on the nerve due to muscle retraction. PMID- 14564277 TI - Electromyographic analysis of internal rotational motion of the shoulder in various arm positions. AB - The electromyographic activity of 9 muscles of the rotator cuff and shoulder girdle was measured in 8 volunteers during active motion and isometric contraction during internal rotation of the shoulder in four positions as follows: neutral rotation with the arm at the side, neutral rotation at 90 degrees abduction in the frontal plane, the zero-position as described by Saha, and the position of the liftoff test from the midlumbar spine as described by Gerber and Krushell. The subscapularis was active at levels of about 44% maximal manual muscle test (MMT) during active internal rotation in the liftoff test position, whereas the activity of the subscapularis decreased remarkably to about 2% to 12% MMT in the other positions. The subscapular is exceptionally important as an internal rotator in the liftoff test position. During resistive motion at the zero-position, the activity level of the subscapularis (81.7%) was higher than that of other muscles. These results indicate that internal rotation at the zero-position may selectively exercise the subscapularis muscle. PMID- 14564278 TI - Syringomyelic neuropathic ulcer of the elbow: treatment with an external fixator. PMID- 14564279 TI - A case of osteocartilaginous mass involving the coronoid process of the ulna: solitary osteochondroma or dysplasia epiphysealis hemimelica? PMID- 14564280 TI - A case of locking shoulder caused by longitudinal rotator cuff tear. PMID- 14564281 TI - Unstable nonunion of the scapula: a case report. PMID- 14564282 TI - Subdeltoid approach for removal of large soft-tissue lesions beneath the deltoid muscle: report of two cases. PMID- 14564283 TI - Posterior shoulder dislocation with lesser tuberosity and scapular spine fractures. PMID- 14564284 TI - Temporomandibular disorder or Eagle's syndrome? A clinical report. AB - This clinical report describes the diagnosis and treatment of a patient under emotional stress with orofacial pain, headaches, and the feeling of a foreign body in the throat. An elongated styloid process at the beginning of the oral pharynx was diagnosed. Although these symptoms could be aspects of Eagle's syndrome, deflective occlusal interferences, tender muscles of mastication, and a clicking temporomandibular joint led to an evaluation for temporomandibular disorder related to malocclusion. An occlusal splint was used to confirm the diagnosis and to alleviate symptoms. Occlusal adjustments were subsequently performed. In a 10-year follow-up, the patient had no complaints. PMID- 14564285 TI - Prosthetic treatment of a patient with facioscapulohumeral muscular dystrophy: a clinical report. AB - Facioscapulohumeral muscular dystrophy syndrome (FSHD) is a rare hereditary myopathy characterized by muscle atrophy and weakness, particularly in the face and upper arms. Patients may also exhibit dental malocclusions. This article presents the prosthodontic treatment for an 18-year old male with FSHD. PMID- 14564286 TI - In vivo fracture resistance of implant-supported all-ceramic restorations. AB - STATEMENT OF PROBLEM: Because of their specific mechanical properties, all ceramic restorations demonstrate a lower fracture resistance than ceramic restorations supported by metal substructures. However, advances have been made in the fabrication of high-strength all-ceramic abutments for anterior implants. No previous study has compared the fracture loads between 2 different all-ceramic abutments restored by glass-ceramic crowns. PURPOSE: The purpose of this in vitro investigation was to quantify the fracture load of implanted-supported Al(2)O(3) and ZrO(2) abutments restored with glass-ceramic crowns. MATERIALS AND METHODS: Two ceramic abutments were tested: an Al(2)O(3) abutment (CerAdapt) and a ZrO(2) abutment (Wohlwend Innovative). The abutments (n = 10) were placed on Branemark dental implants and prepared for restoration with glass-ceramic crowns (IPS Empress). After fabrication, in accordance with the manufacturer's guidelines, the crowns were bonded to the all-ceramic abutments with a dual-polymerizing resin luting agent. The fracture loads (N) were determined by force application at an angle of 30 degrees by use of a computer-controlled universal testing device. The data were analyzed with the unpaired t test (alpha=.05). RESULTS: Statistical analysis showed significant differences between both groups (P=.001) of all-ceramic abutments, with mean fracture load values of 280.1 N (+/- 103.1) for the Al(2)O(3) abutments and 737.6 N (+/- 245.0) for the ZrO(2) abutments. CONCLUSION: Within the limitations of this study, both all-ceramic abutments exceeded the established values for maximum incisal forces reported in the literature (90 to 370 N). The ZrO(2) abutments were more than twice as resistant to fracture as the Al(2)O(3)-abutments. PMID- 14564287 TI - Thermal expansion and microstructural analysis of experimental metal-ceramic titanium alloys. AB - Statement of problem Low-fusing porcelains for titanium veneering have demonstrated inferior color stability and metal-ceramic longevity compared to conventional porcelains. PURPOSE: This study evaluated the microstructure and thermal expansion coefficients of some experimental titanium alloys as alternative metallic substrates for low-fusing conventional porcelain. MATERIALS AND METHODS: Commercially pure titanium (CP Ti) and various metallic elements (Al, Co, Sn, Ga, In, Mn) were used to prepare 8 titanium alloys using a commercial 2-chamber electric-arc vacuum/inert gas dental casting machine (Cyclarc). The nominal compositions of these alloys were the following (wt%): I: 80Ti-18Sn-1.5In-0.5Mn; II: 76Ti-12Ga-7Sn-4Al-1Co; III: 87Ti-13Ga; IV: 79Ti-13Ga 7Al-1Co; V: 82Ti-18In; VI: 75.5Ti-18In-5Al-1Co-0.5Mn; VII: 85Ti-10Sn-5Al; VIII: 78Ti-12Co-7Ga-3Sn. Six rectangular wax patterns for each test material (l = 25 mm, w = 3 mm, h = 1 mm) were invested with magnesia-based material and cast with grade II CP Ti (control) and the 8 experimental alloys. The porosity of each casting was evaluated radiographically, and defective specimens were discarded. Two cast specimens from CP Ti and alloys I-VIII were embedded in epoxy resin and, after metallographic grinding and polishing, were studied by means of scanning electron microscopy and wavelength dispersive electron probe microanalysis. One specimen of each material was utilized for the determination of coefficient of thermal expansion (CTE) with a dilatometer operating from room temperature up to 650 degrees C at a heating rate of 5 degrees C/minute. RESULTS: Secondary electron images (SEI) and compositional backscattered electron images (BEI-COMPO) revealed that all cast specimens consisted of a homogeneous matrix except Alloy VIII, which contained a second phase (possibly Ti(2)Co) along with the titanium matrix. The results showed that the coefficient of thermal expansion (CTE) varied from 10.1 to 13.1 x 10(-6)/ degrees C (25 degrees -500 degrees C), depending on the elemental composition. CONCLUSION: The CTE of titanium can be considerably changed by alloying. Two-phase alloys were developed when alloying elements were added in concentrations greater than the maximum solubility limit in alpha titanium phase. PMID- 14564288 TI - Variability of porcelain color reproduction by commercial laboratories. AB - STATEMENT OF PROBLEM: Many investigations in the field of metal ceramics have examined materials, manipulative variables, and the relationship of these factors to a restoration's color. However, the effect of the artistic component of restoration fabrication is not known. PURPOSE: The purpose of this study was to determine through instrumental colorimetry the variability in color reproduction for metal ceramic crowns fabricated by commercial dental laboratory technicians. MATERIALS AND METHODS: Fifty metal ceramic crowns were fabricated on standardized metal frameworks to the same shade specifications by 5 commercial dental laboratories (n=10). Laboratory prescriptions requested that the technician match the shade and translucency of a provided Vita Lumin A3.5 shade tab. Technicians used the porcelain and technique of their own selection to match the tab. Color differences were determined by use of a colorimeter between crowns and the prescribed shade tab at middle and incisal sites. Analysis of variance was used to determine whether differences in color reproduction existed among laboratories. Where statistically significant interactions existed, the Tukey honestly significant difference test was used to determine significant differences between laboratories according to sites (alpha=.05). RESULTS: Color reproduction was significantly different (P <.0001) among laboratories for both sites. Mean color difference from shade tabs ranged from 3.5 to 11.1 DeltaE units. All laboratories were better at matching shades in the incisal third of the crown. CONCLUSION: Within the limitations of this study, the ability to reproduce the color of the target shade tab differed among laboratories. Most crowns fabricated by the laboratories in this study, when compared to the prescribed shade tab, were above the clinical threshold for an acceptable shade match under intraoral conditions (DeltaE 3.7). PMID- 14564289 TI - Effects of two preparation designs and pontic distance on bending and fracture strength of fiber-reinforced composite inlay fixed partial dentures. AB - STATEMENT OF PROBLEM: Joint fractures observed in Targis/Vectris inlay adhesive fixed restorations may be related to the preparation design. PURPOSE: This in vitro study investigated the effects of the proximal tooth preparation design and the pontic distance on the fracture strength and the amount of bending of fiber reinforced inlay adhesive fixed partial dentures. MATERIALS AND METHODS: Forty extracted premolars and 40 molars were embedded in a PMMA resin to represent a premolar and molar mesiodistal separation distance of 7 mm and 11 mm, respectively. Two preparation designs were used (proximal box and tub-shaped). The sample size was 10 for each group. Fiber-reinforced inlay adhesive fixed partial dentures were fabricated by use of the Targis/Vectris system and luted adhesively to the teeth with Variolink luting agent. A vertical force was loaded to the center of the fixed partial dentures at a crosshead speed of 1 mm/min. The initial bending (mm) prior to fracture was evaluated by measuring the distance the test rod moved from a 10 N preload to fracture. The differences in the mean fracture strength and the average amount of bending as a function of the preparation designs and pontic distances were compared by use of a 2-way analysis of variance (alpha=.05). The specimens were examined optically for the type of failure with a stereomicroscope. The fracture surface of the specimens was examined by scanning electron microscopy, and radiography was used to investigate the surface morphological features at the failure sites and to determine the fracture mode. A chi-square test was used to identify the differences in the debonding rates between the types of preparation designs and the pontic distance (alpha=.05). RESULTS: The mean fracture strength and the standard deviation of the fiber-reinforced inlay retained adhesive fixed partial denture group was 1368+/-212 N for the 7-mm tub group, 885+/-109 N for the 11-mm tub group, 1779+/ 317 N for the 7-mm box group, and 1336+/-281 N for the 11-mm box group. The fracture strength was significantly higher in the 7-mm pontic distance (P<.001) and for the box-shaped tooth preparation (P<.001). The amount of bending was significantly greater in the 7-mm pontic distance (P=.025) and the box-shaped tooth preparation (P=.002). Debonding was observed only in premolar teeth and tub shaped design groups. CONCLUSION: The box-shaped tooth preparation may be considered for restoration of a missing single posterior tooth with fiber reinforced inlay adhesive fixed partial dentures. PMID- 14564290 TI - The effect of surface moisture on detail reproduction of elastomeric impressions. AB - STATEMENT OF PROBLEM: Monophase and dual-viscosity impression techniques are available with little knowledge of which one might render better quality under wet and dry surface conditions. PURPOSE: The purpose of this study was to determine whether type of material, viscosity selection, and presence of moisture affect detail reproduction of elastomeric impressions. MATERIALS AND METHODS: Single-viscosity systems were polyether (Impregum Penta) and vinyl polysiloxanes (President MonoBody, Extrude MPV, and Aquasil). Dual-viscosity systems included polyether (Impregum Penta/Permadyne Garant) and vinyl polysiloxanes (Dimension Penta H/Dimension Garant L, Extrude Extra/Extrude Wash, and Aquasil/Aquasil LV). Impressions were made of a surface analyzer calibration standard possessing a uniform "saw-tooth" pattern with a mean roughness (Ra) of 2.87 mum, which was one fourth of the peak-to-valley height. Each of the 8 impression groups was subjected to dry (control) and wet conditions. The wet condition consisted of 3 mL of distilled water applied to the surface of the standard but allowed to escape during the procedure. Eighty impressions were made, 5 for each test group. After setting, the surface of each impression was scanned at 5 locations using a Surfanalyzer 4000. A 3-factor ANOVA and Student-Newman-Kuels test were used to analyze the data (alpha=.05). RESULTS: There were significant differences between polyether and vinyl polysiloxane materials, dual and monophase techniques, and the 2 surface conditions (P<.05). Cross-product interactions were not significant, allowing comparison of mean values for each factor. The mean Ra for single viscosity was 2.21 mum versus 1.67 mum for dual viscosity; polyether was 2.12 mum versus 1.89 mum for addition silicone; and under dry conditions, the mean was 2.04 mum versus 1.86 mum for wet conditions. CONCLUSION: Single viscosity systems reproduced the standard saw-tooth pattern better than the dual viscosity systems, as did polyether impression materials compared to addition silicones. Moisture led to a lower Ra or less detail compared to dry conditions. PMID- 14564291 TI - Dimensional accuracy and surface detail reproduction of two hydrophilic vinyl polysiloxane impression materials tested under dry, moist, and wet conditions. AB - STATEMENT OF PROBLEM: A major limitation of vinyl polysiloxane (VPS) impression materials is their hydrophobicity. There are 2 aspects to this problem, the wettability of the polymerized impression by dental gypsum materials and the ability of the unpolymerized material to wet intraoral tissues. To address this problem, manufacturers have added surfactants and labeled the new products as hydrophilic vinyl polysiloxane. PURPOSE: The purpose of this investigation was to compare dimensional accuracy and surface detail reproduction of 2 hydrophilic VPS impression materials, when used under dry, moist, and wet conditions. MATERIALS AND METHODS: A total of 102 impressions were made of stainless steel metal dies similar to those described in American Dental Association (ADA) specification 19. The dies had 2 vertical and 3 horizontal lines inscribed on their superior surfaces. Impressions were made under dry, moist, and wet conditions. Dimensional accuracy was measured by comparing the average length of the middle horizontal line in each impression to the same line on the metal die, by use of a measuring microscope with an accuracy of 0.001 mm. A 2-way analysis of variance and least significant difference post hoc test were used to compare mean dimensional changes (alpha=.05). Surface detail reproduction was evaluated in 2 ways: (1) by use of criteria similar to ADA specification 19 for detail reproduction, continuous replication of at least 2 of the 3 horizontal lines, and (2) by use of a method developed for this study that categorized the impressions as satisfactory or unsatisfactory based on their surface characteristics: presence of pits, voids, or roughness. Pearson chi(2) (alpha=.05) was used to compare detail reproduction results. RESULTS: Conditions (dry, moist, and wet) did not cause significant adverse effects on the dimensional accuracy of either material. The mean dimensional change and SD were 0.005% +/- 0.002% or less. With both surface detail analyses, dry, moist, and wet conditions had a significant effect on the detail reproduction of both materials (P<.05). Only under dry conditions did both impression materials continuously replicate at least 2 of the 3 horizontal lines 100% of the time. Under moist conditions, 82% of the Aquasil impressions and 100% of the Reprosil impressions were judged satisfactory, while under wet conditions, only 47% Aquasil and 11% Reprosil impressions were satisfactory. With the additional surface detail characterization, only under dry conditions were impressions produced with clinically acceptable surface quality (Aquasil 77% and Reprosil 100% satisfactory). CONCLUSIONS: Dimensional accuracy of both materials tested was well within ADA standards. Best surface detail results were obtained only under dry conditions for both materials. PMID- 14564292 TI - Occlusion: reflections on science and clinical reality. AB - The last 50 years have seen progress in emphasizing scientific evidence as a basis for dental practice, including occlusal therapy. Although a proper understanding of the contributory role of occlusion to temporomandibular disorders should not be minimized, the importance of occlusion in other areas of dental education and practice should not be overlooked. The primary objective of this article is to discuss the nature of this problem as it relates to the duality of science and clinical reality in the evidence-based paradigm, information transfer, quality of evidence, clinical trials, and clinical aspects of occlusion. Some suggested solutions for the problem and thoughts on past and future perspectives of occlusion are expressed in context of inquiry. PMID- 14564293 TI - The effect of length and concentration of glass fibers on the mechanical properties of an injection- and a compression-molded denture base polymer. AB - STATEMENT OF PROBLEM: Fiber-reinforcement has been used to overcome the mechanical limitations of denture base polymers. One major difficulty in the use of fiber reinforcement has been the addition of fibers during conventional processing methods. PURPOSE: This study evaluated the effect of various lengths and concentrations of chopped E-glass fiber-reinforcement on the transverse strength, modulus of elasticity, and impact strength of injection and compression molded polymethyl methacrylate based denture base polymer. MATERIALS AND METHODS: Test specimens (n=10) of 4-, 6-, and 8-mm fiber length and 1%, 3%, and 5% weight fiber concentrations were prepared with either an injection or a compression molded processing method. Denture base polymer specimens without any fiber reinforcement were used as control for both processing methods. Transverse strength test specimens (65 x 10 x 2.5 mm) were stored in water bath at 37 degrees C for 2 weeks. The transverse strength (MPa) and modulus of elasticity (GPa) was measured with the 3-point bending test. Impact strength (kJ/m(2)) test specimens (60 x 7.5 x 4 mm) were tested with the Charpy-type pendulum impact test setup. The data were analyzed with multifactorial analysis of variance and Tukey post hoc tests (alpha=.05). RESULTS: Injection-molded fiber-reinforced groups showed significantly higher transversal strength, elastic modulus, and impact strength compared with compression-molded groups (P <.001). In the injection molded groups, fiber concentration increased all mechanical properties tested (P <.05), but fiber length only increased transverse strength and modulus of elasticity (P <.05). In the compression molded groups, fiber concentration affected modulus of elasticity and impact strength significantly (P <.05), but fiber length did not show any significant effect on the mechanical properties tested (P >.05). CONCLUSION: The transverse strength, elastic modulus and impact strength of injection-molded denture base polymer increased significantly with the use of chopped E-glass fibers, whereas the effect was not significant with the compression-molded polymer. PMID- 14564294 TI - Effect of complete dentures on dynamic measurement of changing head position: A pilot study. AB - STATEMENT OF PROBLEM: Complete dentures contribute significantly to the facial esthetics of edentulous patients. However, information as to the effect of complete dentures on the natural position of the head is limited. PURPOSE: The purpose of this pilot study was to evaluate the immediate and 30-day effect of wearing complete dentures on the dynamic natural head position measured during walking. MATERIALS AND METHODS: The sample consisted of a volunteer group of 16 patients, 8 women and 8 men, who received new complete dentures. The ages of the subjects ranged from 45 to 64 years (mean=52 years). Dynamic measurement of head posture was carried out by a specially constructed inclinometer device. Each subject in turn was fitted with the inclinometer system and instructed to walk in a relaxed manner for 5 minutes. The data, measured as degrees, were stored in a pocket data logger. This procedure was repeated before insertion of dentures (T1), immediately after insertion of dentures (T2), and 30 days after insertion of dentures (T3). Stored dynamic head posture data were transferred to computer for analysis. The means of the measurements were statistically compared with Friedman and following Wilcoxon tests (alpha =.05). RESULTS: Twelve of 16 (75%) subjects showed an average of 4.6 degrees of cranial extension immediately after insertion of dentures. Six (37.5%) subjects showed an average of 6.4 degrees of cranial flexion, and 8 (50%) subjects showed an average of 5.2 degrees of cranial extension at T3 relative to the T1 measurement. Dynamic head posture measurements of the other 2 subjects remained unchanged. There were significant differences between different measurements of dynamic head posture positions (P<.025). However, only the T1 and T2 measurements were significantly different (P<.015). CONCLUSION: The findings indicate that the statistically significant average extension 4.6 degrees in subjects immediately after insertion of complete dentures was not stable after a 30-day evaluation period and did not produce any statistically significant change. The overall effect of wearing dentures was an irregular flexion or extension pattern on dynamic head posture. PMID- 14564295 TI - A technique to determine a desired preparation axial inclination. AB - The guidelines recommended in the literature for the convergence angle of a crown preparation vary from 3 to 24 degrees. There is a lack of guidelines on techniques to achieve a specific axial inclination. The purpose of this article was to present a practical technique, with a diamond rotary cutting instrument of known axial inclination, to determine the diamond rotary cutting instrument angulations required to achieve the desired axial inclination of a preparation. PMID- 14564296 TI - The modification of interim cylinders for the fabrication of cement-retained implant-supported provisional restorations. AB - This article describes a technique for modification of metal interim cylinders and their use in the fabrication of cementable implant-supported provisional restorations. This chairside technique allows for direct provisionalization of single or multiple implants during or after second-stage surgery, especially when there is a high esthetic demand. PMID- 14564297 TI - Use of transparent polyvinylsiloxane to replicate gingival peri-implant soft tissue. AB - The fabrication of an esthetic implant-supported restoration often necessitates consideration of the surrounding gingival architecture. Procedures for generating a definitive cast that accurately details the peri-implant soft tissue with a resilient material have been described. Currently used gingival replicating materials are usually pink and opaque, making it difficult or impossible for the dentist and laboratory technician to visually confirm complete seating of components subgingivally on laboratory analogs without altering the cast. This article describes a technique for fabricating a definitive cast using a transparent polyvinylsiloxane material to replicate the peri-implant gingiva. PMID- 14564298 TI - Chair-side addition of cast clasp to removable partial denture after abutment extraction. PMID- 14564299 TI - Inflammation as a therapeutic target: a unique role for abciximab. AB - Vascular inflammation is a central pathogenic mechanism for both acute coronary syndromes and the vascular response to injury after percutaneous coronary intervention. The magnitude of vascular inflammation has been correlated with adverse late clinical outcomes (death, myocardial infarction, recurrent ischemia, restenosis). Vascular inflammation is also increased in patients with diabetes mellitus. Many adjunctive pharmacotherapies used in the treatment of acute coronary syndromes or during percutaneous coronary intervention have anti inflammatory effects, which are distinct from their perceived primary mechanism of action. Data in support of the anti-inflammatory effects of abciximab are presented and the role that these effects may play in modulating atherosclerotic plaque stability and late clinical outcomes is discussed. Vascular inflammation represents the "final common pathway" for many disease processes and thus represents the "ultimate therapeutic target" for pharmacologic intervention. PMID- 14564300 TI - Loss of bone marrow-derived vascular progenitor cells leads to inflammation and atherosclerosis. AB - BACKGROUND: Aging represents the most powerful risk for the development of atherosclerosis and atherosclerotic thromboembolic complications. Yet, the mechanism by which aging affects the arterial wall and its deterioration has remained essentially uncharacterized. FINDINGS: Chronic injuries to the arterial wall contribute to the development of atherosclerosis. However, it is important to note that a complex repair system that involves both local and bone marrow derived cells maintains arterial homeostasis and integrity. With this review, we explain how the age-dependent failure of the bone marrow to produce vascular progenitor cells responsible for such arterial repair--an inability that results from the impact of a lifetime of risk factors such as hyperlipidemia--drives atherosclerosis and its thromboembolic complications. As a consequence of such failure, the normal processes of arterial wall repair and rejuvenation are impaired. The disequilibrium that ensues between injury of the arterial wall and repair leads to atherosclerotic inflammation and consequent thromboembolic complications. CONCLUSION: The bone marrow and derived progenitor cells represent key regulators of atherosclerosis, and progress in the prevention and treatment of atherosclerosis and its thromboembolic complications will need to take into account this new dimension for the disease process. PMID- 14564301 TI - Drug-eluting stents and glycoprotein IIb/IIIa inhibitors: combination therapy for the future. AB - BACKGROUND: Although coronary stenting has improved the results of coronary interventions compared to coronary angioplasty alone, in-stent restenosis remains a significant limitation of this procedure. Drug-eluting stents with or without glycoprotein IIb/IIIa inhibitor therapy represent an additional advance in the evolution of this strategy. METHODS: We review the currently available trials comparing studies of non-drug-eluting and drug-eluting stents using sirolimus and paclitaxel agents and their derivatives. RESULTS: Ten studies are available that compare drug-eluting to traditional non-drug-eluting stents. A variety of antiplatelet regimes have been used. The majority of these studies are in the process of being published. No head-to-head studies comparing different drug eluting stents are available. CONCLUSIONS: Drug-eluting stents using sirolimus and paclitaxel in combination with enhanced antiplatelet strategies represent an important advantage over non-drug-eluting stents for the reduction of in-stent restenosis. The rate at which drug-eluting stents are adapted into widespread practice depends heavily on whether they are safe, efficacious, and cost effective in various clinical settings. PMID- 14564302 TI - Glycoprotein IIb/IIIa inhibitors in patients with unstable angina/non-ST-segment elevation myocardial infarction: appropriate interpretation of the guidelines. AB - In 2002, the American College of Cardiology and the American Heart Association published an update to their guidelines for the management of patients with unstable angina and non-ST-segment elevation myocardial infarction. These revised guidelines make specific recommendations regarding the use of glycoprotein IIb/IIIa inhibitors. This article briefly reviews the evidence supporting the use of glycoprotein IIb/IIIa inhibitors in unstable angina and non-ST-segment elevation myocardial infarction, before moving on to discuss interpretation of these new guidelines. PMID- 14564304 TI - Matters of the heart: assessing the cardiovascular safety of new drugs. PMID- 14564303 TI - Direct thrombin inhibitors for non-ST-segment elevation acute coronary syndromes: what, when, and where? AB - BACKGROUND: Unfractionated heparin has been the cornerstone of antithrombin therapy in the treatment of non-ST-segment elevation acute coronary syndromes for more than a decade. Several new anticoagulants have emerged in recent years and have been studied extensively in patients with unstable coronary syndromes and in the percutaneous coronary intervention setting. METHODS: Direct thrombin inhibitors comprise a family of agents with promising properties that offer several potential advantages over unfractionated heparin. Hirudin has been studied in patients with ST-elevation myocardial infarction, non-ST-elevation coronary syndromes, and coronary angioplasty. Bivalirudin has been studied in patients undergoing percutaneous coronary revascularization, with very promising efficacy and safety profile compared with unfractionated heparin. RESULTS: The clinical trials of direct thrombin inhibitors in non-ST-elevation acute coronary syndromes and coronary angioplasty are reviewed. PMID- 14564305 TI - Pharmaceutical advertising versus research spending: are profits more important than patients? PMID- 14564306 TI - Choice of treatment: outcomes and treatment goals. PMID- 14564307 TI - Of millimeterology and cannibalism: comparative antihypertensive efficacy of angiotensin receptor blockers. PMID- 14564308 TI - Screening for structural heart disease: time to stop listening and start looking. PMID- 14564309 TI - Assessing risk for coronary heart disease: beyond Framingham. PMID- 14564310 TI - The nested case-control study in cardiology. AB - BACKGROUND: The nested case-control study is an efficient epidemiological design whereby a case-control approach is employed within an established cohort. The large number of recent prospective studies and randomized trials conducted in cardiology provide cohorts within which the nested case-control approach is increasingly used. METHODS: This paper describes the design of the nested case control study, and evaluates its role in cardiology by reviewing all such studies indexed in Medline from 1966 to 2000. The example of homocysteine is used to illustrate how discrepancies between results of nested case-control and case control studies played an important role in the decisions and recommendations of national and international organizations. RESULTS: Seventy-seven nested case control studies in cardiology were reviewed. The number of studies per year has been increasing since the first publication in 1987. The majority (96%) of studies evaluated potential risk factors for cardiovascular disease while the remainder evaluated drugs with cardiac adverse effects. In studies of homocysteine and coronary artery disease, nested case-control studies did not confirm the strong association suggested by early case-control studies that may have been influenced by bias (eg, selection, publication, or reverse causality). This led national and international organizations to advise against routine screening. CONCLUSIONS: The nested case-control study is increasingly used to study causal relationships in cardiology. The large cohorts of cardiac patients created by prospective studies, clinical trials, and administrative databases should be exploited using this methodology to assess potential cardiac risk factors and other causal relationships that cannot be studied in randomized trials. PMID- 14564311 TI - Selective COX-2 inhibition and cardiovascular effects: a review of the rofecoxib development program. AB - See related Editorials on pages 561 and 563. Cyclo-oxygenase-2 (COX-2) inhibitors appear to alter the balance of vasoactive eicosanoids (prostacyclin and thromboxane) and to suppress the inflammatory mediators implicated in the progression of atherogenesis and ischemic myocardial injury. Neutral, harmful, and beneficial cardiovascular (CV) effects have all been postulated to result from these changes. Investigations conducted with rofecoxib, a selective COX-2 inhibitor, have substantially contributed to our understanding of this scientific area. Rofecoxib had little or no effect on platelet aggregation or platelet derived thromboxane synthesis but reduced systemic prostacyclin synthesis by 50% to 60%. These findings prompted extensive analyses of CV thrombotic events within the rofecoxib development program. Among 5435 osteoarthritis trial participants, similar rates of CV thrombotic events were reported with rofecoxib, placebo, and comparator, nonselective NSAIDs (ibuprofen, diclofenac, and nabumetone). In the VIGOR gastrointestinal outcomes trial of >8000 patients, naproxen (an NSAID with aspirin-like sustained antiplatelet effects throughout its dosing interval) was associated with a significantly lower risk of CV events than was rofecoxib. A subsequent pooled analysis from 23 studies (including VIGOR) encompassing multiple disease states and including more than 14,000 patient-years at risk also demonstrated that rofecoxib was not associated with excess CV thrombotic events compared with either placebo or nonnaproxen NSAIDs. Again, naproxen appeared to be the outlier, suggesting a cardioprotective benefit of naproxen. Finally, among the predominantly elderly, male population participating in Alzheimer trials, both rofecoxib- and placebo-treated patients had similar rates of CV thrombotic events. The totality of data is not consistent with an increased CV risk among patients taking rofecoxib. PMID- 14564312 TI - Changing the model of care for patients with acute coronary syndromes. AB - Acute coronary syndromes (ACS) represent a major cause of morbidity and mortality for patients with cardiovascular disease, but evidence-based therapies shown to improve outcomes for ACS are often underused in clinically eligible patients. Although clinical practice guidelines have been developed to provide standards for the diagnosis and treatment of patients with ACS and to provide physicians with a framework for clinical decision-making, multiple obstacles have hindered their implementation and questions remain about the applicability of guidelines for diverse clinical situations. Systematic reviews of quality-improvement studies have shown that multifaceted approaches using targeted educational interventions, creation of quality standards, and regular performance feedback are needed to ensure sustained improvements in care. Approaches to quality improvement thus are being redirected to focus on multidisciplinary collaborations designed to improve the entire process of care for patients with ACS. Multiple large observational registries and quality-improvement initiatives now are capturing data regarding adherence to practice guidelines and contemporary patterns of care for ACS. This comprehensive evaluation of ACS treatment will help guide efforts designed to promote evidence-based care and ultimately determine the effect of widespread implementation of practice guidelines on clinical outcomes. The shifting model of care for ACS therefore suggests that quality improvement and monitoring of adherence to practice guidelines should be considered components of optimal clinical practice. PMID- 14564313 TI - Effects of eicosapentaenoic acid on cardiovascular events in Japanese patients with hypercholesterolemia: rationale, design, and baseline characteristics of the Japan EPA Lipid Intervention Study (JELIS). AB - HYPOTHESIS: The principle aim of the current study is to test the hypothesis that the long-term use of highly purified EPA (eicosapentaenoic acid: 1800 mg/day), in addition to HMG-CoA reductase inhibitor, is effective in preventing cardiovascular events in Japanese patients with hypercholesterolemia. BACKGROUND: Epidemiological and clinical evidence suggest that intake of long-chain polyunsaturated n-3 fatty acids (PUFAs), which are abundant in fish, might have a significant role in the prevention of coronary artery disease, as marine PUFAs have multiple biological functions through lipid-dependent and lipid-independent mechanisms. METHODS: The Japan EPA Lipid Intervention Study (JELIS) is a prospective, randomized, open-label, blinded end point trial including both primary and secondary prevention strata, with a maximum follow-up of 5 years. Its main purpose is to examine the clinical effectiveness of EPA oil given as an additional treatment to patients taking HMG-CoA reductase inhibitors for hypercholesterolemia. A primary end point is major coronary events: sudden cardiac death, fatal and nonfatal myocardial infarction, and unstable angina pectoris including hospitalization for documented ischemic episodes, and events of angioplasty/stenting or coronary artery bypass grafting. Secondary end points include all-cause mortality, stroke, peripheral artery disease, and cancer. Baseline study composition comprises 15,000 participants (4204 men and 10,796 women) in the primary prevention stratum and 3645 (1656 men and 1989 women) in the secondary stratum. The minimum age is 40 years for men, women are required to be postmenopausal, and all patients must be < or =75 years of age. The mean age of participants is 61 years, and 69% are female. The schedule for plasma fatty acid composition measurement is as follows: at baseline, at 6 month, and yearly thereafter. The mean baseline total and low-density lipoprotein cholesterol levels were 275 mg/dL (7.1 mmol/L) and 180 mg/dL (4.6 mmol/L). RESULTS: Results are expected in 2005. CONCLUSION: JELIS is a large clinical trial that will evaluate whether EPA can make an additional improvement in mortality and morbidity of coronary artery disease beyond that of HMG-CoA reductase inhibitor treatment. PMID- 14564314 TI - Revascularization in patients with coronary artery disease, left ventricular dysfunction, and viability: a meta-analysis. AB - BACKGROUND: The effects of viability status and treatment allocation on long-term mortality in patients with left ventricular dysfunction and coronary artery disease have not been determined. Several observational studies with significant limitations have addressed this issue, and a recent meta-analysis has attempted to combine these results to increase statistical power. However, the analysis did not test for an interaction between viability status and treatment type, and included extraneous studies. We provide an alternate meta-analysis of this primary literature, utilizing interaction statistical methodology on relevant data and factoring in multiple limitations. METHODS: We examined papers from this prior meta-analysis examining viable and nonviable patients undergoing surgical or medical therapy. We determined an interaction odds ratio for each study and used an empirical Bayes random-effects model to obtain a combined interaction odds ratio that was tested for statistical significance. We compared our results against an interaction odds ratio we estimated from the primary studies included in the previous meta-analysis. RESULTS: Nine relevant studies with 1244 patients and 172 events were identified that utilized all 4 treatment/viability subsets. The interaction odds ratio was 2.76 (P =.0176, 95% CI 1.19-6.38), 2.5 times lower than our estimated interaction odds ratio of 7.27 for the prior meta-analysis. CONCLUSIONS: We found a markedly reduced but statistically significant interaction between viability status and treatment allocation. However, numerous limitations in the primary studies and the application of meta-analysis along with significant improvements in medical therapies render a randomized controlled trial necessary to reach a definitive conclusion to this critical question. PMID- 14564315 TI - Combining enoxaparin and glycoprotein IIb/IIIa antagonists for the treatment of acute coronary syndromes: final results of the National Investigators Collaborating on Enoxaparin-3 (NICE-3) study. AB - BACKGROUND: In high-risk patients with acute coronary syndromes (ACS), there have been concerns relating to the safety of using low molecular weight heparins (LMWH) in combination with a glycoprotein (GP) IIb/IIIa antagonist, and the continued use of LMWH in patients brought to the cardiac catheterization laboratory for percutaneous coronary intervention (PCI). METHODS: The National Investigators Collaborating on Enoxaparin-3 (NICE-3) study was an open-label observational study of enoxaparin in combination with any 1 of 3 available GP IIb/IIIa antagonists in patients presenting with non-ST-elevation ACS. The primary end point was the incidence of major bleeding not related to coronary artery bypass graft (CABG) surgery. Data were also recorded on the incidence of death, myocardial infarction (MI), and urgent revascularization for repeat ischemia. RESULTS: A total of 671 patients with validated data were treated with enoxaparin; 628 of these patients also received a GP IIb/IIIa antagonist (tirofiban, n = 229; eptifibatide, n = 272; abciximab, n = 127); 283 of 628 underwent percutaneous coronary intervention (PCI). The 30-day incidence of non CABG major bleeding was 1.9%, and was not significantly higher than a prespecified historical control rate of 2.0%. Outcome events included death (1.0% at hospital discharge and 1.6% at 30 days), MI (3.5% and 5.1%, respectively), and urgent revascularization (2.7% and 6.8%, respectively). CONCLUSIONS: The safety of enoxaparin plus a GP IIb/IIIa antagonist was comparable to that of unfractionated heparin plus a GP IIb/IIIa antagonist, as reported in other recent major trials. Patients undergoing PCI can be safely managed with enoxaparin and a GP IIb/IIIa antagonist, without supplemental use of unfractionated heparin. PMID- 14564316 TI - Cardiac pathologic findings reveal a high rate of sudden cardiac death of undetermined etiology in younger women. AB - BACKGROUND: Between 1989 and 1998 there was a 21% increase in estimated sudden cardiac death among US women aged 35 to 44 years. In contrast, the sudden cardiac death rate in age-matched men showed a decreasing trend (-2.8%). Due to under representation of younger adults in published autopsy series, etiologies of sudden cardiac death merit further investigation. METHODS: We reviewed autopsy and detailed cardiac pathologic findings in younger women (age 35-44 years) from a 270-patient, 13-year (1984-1996) autopsy series of sudden cardiac death, and performed comparisons with findings in age-matched men. RESULTS: Women aged 35 to 44 years constituted 32% of all women in the series compared to men, who constituted 24% of total men (P =.004 vs women). A presumptive cause of sudden cardiac death could not be determined in 13 women (50%). Among women, 6 cases (22%) had significant coronary artery disease. Findings in others included coronary artery anomalies (n = 3), myocarditis (n = 2), hypertrophic cardiomyopathy (n = 1), coronary artery dissection (n = 1) and accessory pathway (n = 1). In younger men, a presumptive cause of sudden cardiac death remained undetermined in only 24% (P =.025 vs younger women), and coronary artery disease accounted for 40% of cases. CONCLUSIONS: In younger women, despite autopsy and detailed cardiac pathologic examination, an attributable cause of sudden cardiac death was not determined in 50% of cases; a 2-fold increase compared to men of the same age. Given the dynamic and multifactorial nature of sudden cardiac death, comprehensive population-based investigations are likely to be necessary to further investigate this unexpected sex-based disparity. PMID- 14564317 TI - Does 24-hour ST-segment resolution postfibrinolysis add prognostic value to a Q wave? An ASSENT 2 electrocardiographic substudy. AB - BACKGROUND: Both ST resolution and Q-wave development postfibrinolysis provide important prognostic insights in patients with acute myocardial infarction (MI). However, the relative contributions of these 2 factors to risk assessment have not been examined prospectively. METHODS AND RESULTS: ST resolution and Q development were evaluated 24 to 36 hours (24-36 h) postfibrinolysis in ASSENT-2: 13,100 out of 16,949 patients who had both baseline and 24-36 h electrocardiograms free of confounders (left bundle branch block, ventricular rhythm, reinfarction before 24-36 h electrocardiograms) were included in this analysis. Q-wave MI evolved in 10,466 patients (79.9%) and 2634 patients (20.1%) had non-Q-wave MI at 24-36 h postfibrinolysis. Mortality rates at 1-year were 7.0% for patients with Q-wave MI and 5.8% for non-Q-wave MI patients, respectively (P =.046). Patients with Q-wave MI versus those without were less likely to have complete ST-segment resolution (49.1% vs 59.1%) and more likely to have partial (37.1% vs 27.8%) or no resolution (13.8% vs 13.1%) at 24 to 36 hours postfibrinolysis (P <.001). Mortality rates at 1 year for Q-wave MI with complete, partial, and no resolution were 5.2%, 8.1%, and 10.1%, respectively (P <.001), and for non-Q-wave MI with complete, partial, and no resolution were 4.5%, 7.6%, and 8.0% (P =.003). CONCLUSION: These results demonstrate the additional prognostic significance of ST-segment resolution to Q-wave development at 24 to 36 hours after fibrinolysis. PMID- 14564318 TI - Impact of angina burden and other factors on treatment satisfaction after acute coronary syndromes. AB - PURPOSE: Although of great importance to clinicians, hospitals, and health care systems, little is known about factors that influence treatment satisfaction after acute coronary syndromes (ACS). The objective of this study was to identify potentially modifiable factors associated with treatment satisfaction 7 months after ACS. METHODS: The study population included 1957 patients with ACS who were enrolled in the multicenter, prospective Veterans' Health Administration Access to Cardiology Study. The primary outcome was treatment satisfaction 7 months after ACS as measured by the Seattle Angina Questionnaire. Multivariable regression models were developed to determine the association between treatment satisfaction and patient characteristics, physician-patient communication, and current angina frequency. RESULTS: Patient characteristics associated with reduced treatment satisfaction included a history of depression, atrial fibrillation, prior heart surgery, arthritis, hypertension, younger age, and a discharge diagnosis of unstable angina (as opposed to myocardial infarction). After adjusting for patient characteristics, patient-reported inability to reach one or more of their physicians (OR, 2.40; 95% CI, 1.47 to 3.91), being given confusing information (OR, 3.48; 95% CI, 2.08 to 5.83), and poor overall communication with one or more of their physicians (OR, 4.94; 95% CI, 2.93 to 8.34) were all associated with reduced satisfaction. Finally, after adjustment for both patient characteristics and physician communication, weekly (OR, 3.52; 95% CI, 2.28 to 5.45) and daily angina (OR, 3.88; 95% CI, 2.23 to 6.75) were associated with worse treatment satisfaction. CONCLUSIONS: Current angina symptoms and aspects of physician communication are independently associated with treatment satisfaction after ACS. These results suggest that treatment satisfaction may be improved through better communication and better control of angina symptoms. PMID- 14564319 TI - Clinical and angiographic characteristics of patients with combined anterior and inferior ST-segment elevation on the initial electrocardiogram during acute myocardial infarction. AB - OBJECTIVE: We evaluated the significance of combined anterior and inferior ST segment elevation on the initial electrocardiogram (EKG) in patients with acute myocardial infarction (AMI) and correlated it with AMI size and left ventricular (LV) function. METHODS: We analyzed admission EKGs of 2996 patients with AMI from the GUSTO-I angiographic substudy and the GUSTO-IIb angioplasty substudy who underwent immediate angiography. In all, we identified 1046 patients with anterior ST elevation (ST-segment elevation in > or =2 of leads V1-V4) and divided them into 3 groups: Group 1, anterior + inferior ST elevation (ST elevation in > or =2 of leads II, III, aVF, n =179); Group 2, anterior ST elevation only (<2 of leads II, III, aVF with ST elevation or depression, n = 447); Group 3, anterior ST elevation + superior ST elevation (ST depression in > or =2 of leads II, III, aVF, n = 420). RESULTS: Cardiac risk factors, prior AMI, prior percutaneous transluminal coronary angioplasty or coronary artery bypass graft, Killip class, and thrombolytic therapy assignment did not differ among the 3 groups. Group 1 patients had greater number of leads with ST elevation compared to Groups 2 and 3 (ST elevation in > or =6 leads 83% vs 22% vs 49%, P =.001). Despite greater ST-segment elevation, Group 1 patients had a lower peak CK level (median baseline peak CK 1370 vs 1670 vs 2381 IU, P =.0001) and less LV dysfunction (median ejection fraction 0.53 vs 0.49 vs 0.45, P =.0001; median number of abnormal chords 21 vs 32 vs 40, P =.0001). Angiographically, Group 1 had 2 distinct subsets of patients with either right coronary artery (RCA) (59%) or left anterior descending coronary artery (LAD) (36%) occlusion. In contrast, the infarct-related artery (IRA) was almost entirely the LAD in Groups 2 and 3 (97%). Further, the site of IRA occlusion in Group 1 was mostly proximal RCA (67%) in the RCA subgroup and mid or distal LAD (70%) in the LAD subgroup. ST segment elevation in lead V1 > or = V3 and absence of progression of ST elevation from lead V1 to V3 on the EKG differentiated IRA-RCA from IRA-LAD in patients with combined anterior and inferior ST elevation. CONCLUSIONS: The AMI size and LV dysfunction in patients with anterior ST elevation is directly related to the direction of ST segment deviation in the leads II, III, aVF; least with inferior ST elevation, intermediate with no ST deviation, and maximal with superior ST elevation (inferior ST depression). Despite greater ST-segment elevation, patients with combined anterior and inferior ST elevation have limited AMI size and preserved LV function. Angiographically, they comprise 2 distinct subsets with either proximal RCA or mid to distal LAD occlusion. A predominant right ventricular and limited inferior LV AMI from a proximal RCA occlusion, or a smaller anterior AMI from a more distal occlusion of LAD may explain their limited AMI size despite greater ST elevation. PMID- 14564320 TI - Arterial compliance adds to conventional risk factors for prediction of angiographic coronary artery disease. AB - BACKGROUND: Arterial compliance is related to left ventricular hypertrophy and risk for cardiovascular disease events; however, its association with coronary artery stenosis remains uncertain. We sought to assess the relation between lower extremity arterial compliance and presence of angiographically defined coronary artery disease. METHODS: Lower extremity arterial compliance was measured with the use of a noninvasive air plethysmography technique in 376 subjects undergoing routine diagnostic coronary angiography. RESULTS: Measures of calf arterial compliance were significantly associated with the presence of one or more stenoses > or =50% compared with no stenoses, even after adjustment for age, sex, smoking, diabetes, hypertension, hypercholesterolemia, and obesity (P =.03). Measures of thigh arterial compliance were also lower in subjects with disease, although this association did not reach statistical significance (P =.07). Receiver operator curves illustrate the incremental predictive ability of calf arterial compliance over and above age, sex, and conventional risk factors. CONCLUSIONS: Lower extremity arterial compliance is associated with presence of significant coronary stenoses in a cardiac catheterization laboratory referral population. This observation lends support for additional efforts to determine the utility of vascular stiffness measures in both clinical and pre-clinical populations to guide treatment and prevention efforts. PMID- 14564321 TI - Effect of platelet glycoprotein IIb/IIIa receptor blockade with tirofiban on adverse cardiac events in women with unstable angina/non-ST-elevation myocardial infarction (PRISM-PLUS Study). AB - BACKGROUND: Previous trials demonstrated the efficacy of platelet glycoprotein IIb/IIIa receptors blockade with tirofiban in reducing acute ischemic events in patients with unstable angina/non-ST-elevation myocardial infarction. Little is known about the effect of tirofiban among women with acute coronary syndromes. OBJECTIVE: We aimed to determine the benefit and safety of tirofiban plus heparin versus heparin alone on cardiac ischemic events among women with unstable angina/non-ST-elevation myocardial infarction. METHODS AND RESULTS: We performed a post hoc analysis of all women enrolled in the PRISM-PLUS trial. At early time points, there appeared to be a reduction of the primary composite end point of death, myocardial infarction, or refractory ischemia among women treated with tirofiban plus heparin (RR, 0.78 and 0.67) compared with women treated with heparin alone. However, at 30 and 180 days, there was no significant reduction of events with the combination therapy of tirofiban plus heparin (treatment-by-sex interaction, P =.05). Death or myocardial infarction was not significantly reduced by the combination therapy among women at all time points. CONCLUSIONS: Although the effects of tirofiban in reducing the primary composite outcome were similar among men and women early in the study, there appeared to be a difference at the later time points. In particular, tirofiban was effective among men, but there was no clear effect among women at 30 and 180 days. PMID- 14564322 TI - Impact of acute hyperglycemia on left ventricular function after reperfusion therapy in patients with a first anterior wall acute myocardial infarction. AB - OBJECTIVE: This study was undertaken to assess the relationship between acute hyperglycemia and left ventricular function after reperfusion therapy for acute myocardial infarction (AMI). METHODS: This study consisted of 529 patients with a first anterior wall AMI who underwent coronary angiography followed by coronary angioplasty or thrombolysis within 12 hours after the onset of chest pain. Plasma glucose was measured at the time of hospital admission. Acute hyperglycemia was defined as plasma glucose >10 mmol/L. RESULTS: Although acute hyperglycemia was associated with both lower acute left ventricular ejection fraction (LVEF) (46% +/- 12% vs 48% +/- 10%, P =.026) and lower predischarge LVEF (51% +/- 15% vs 56% +/- 15%, P =.001), the difference was more pronounced in the latter and the change in LVEF was significantly smaller in patients with acute hyperglycemia (4.8% +/- 11.2% vs 8.0% +/- 13.8%, P =.022). Multivariable analysis showed that there was a significant correlation between plasma glucose and impaired predischarge LVEF, even after adjustment of acute LVEF (r = -0.13, P =.005). Thirty-day mortality tended to be higher in patients with acute hyperglycemia than in patients without (7.1% vs 3.5%, P =.06). Multivariable analysis showed that plasma glucose (per 1 mmol/L increase) was an independent predictor of 30 day mortality after AMI (odds ratio 1.12, 95% CI 1.03-1.22, P =.009). CONCLUSION: Acute hyperglycemia was independently associated with impaired left ventricular function and higher 30-day mortality after AMI. These results may provide a potential explanation for poor outcomes of patients with AMI and acute hyperglycemia. PMID- 14564323 TI - Screening for early detection of cardiovascular disease in asymptomatic individuals. AB - OBJECTIVE: Primary prevention of cardiovascular disease has been aimed at risk factor identification and treatment without efforts to document early cardiovascular disease. The objective of the current study is to screen individuals with vascular and cardiac tests aimed at identifying early abnormalities likely to progress and to measure risk contributors susceptible to therapy. METHODS: A center was established for comprehensive screening of an asymptomatic population with 10 tests designed to detect early vascular and cardiac abnormalities and blood tests to identify potential targets for risk contributor intervention. The first 396 individuals screened in the center have been analyzed. RESULTS: Using a scoring system from 0 (no disease) to 20 (advanced disease), 49% of the population exhibited scores of > or =5 and 39% exhibited scores of > or =6. These scores appear indicative of early disease mandating initiation of or change in medical therapy, which was recommended to the individuals screened and to their primary care physicians. CONCLUSION: The screening tests utilized are effective in uncovering unsuspected early cardiovascular disease in which targeted treatment could be effective in reducing the incidence of cardiovascular events in susceptible individuals. Documentation of the sensitivity and specificity of this approach requires longitudinal study. PMID- 14564324 TI - Primary percutaneous transluminal coronary angioplasty accelerates early myocardial reperfusion compared to thrombolytic therapy in patients with acute myocardial infarction. AB - BACKGROUND: Successful early reperfusion of the infarcted myocardium as indicated by complete resolution of ST-segment elevations has been shown to be associated with an improved outcome in patients with acute ST-elevation myocardial infarction (AMI). The aim of this study was to compare early ST resolution in patients treated with primary percutaneous transluminal coronary angioplasty (PTCA) or thrombolytic therapy for AMI. METHODS: A total of 1379 patients with AMI whose symptoms began <6 hours previously were enrolled in the Evaluation of the Safety and Cardioprotective effects of eniporide in Acute Myocardial Infarction (ESCAMI) trial and treated with primary PTCA (n = 528) or thrombolytic therapy (n = 851). Twelve-lead electrocardiograms (ECG) were obtained at baseline, directly after PTCA and at 90 minutes after the initiation of thrombolytic therapy. RESULTS: There were no differences with respect to clinical or ECG baseline variables between the 2 groups. The time intervals between hospital admission and ECG 2 (obtained 0-30 min after PTCA and 90 min after start of thrombolysis) were 121 +/- 62 minutes in the PTCA group and 137 +/- 57 minutes in the thrombolysis group, respectively. In ECG 2, complete (> or =70%) ST resolution was observed more often in the PTCA treated patients (35 vs 27%, P =.003). The incidence of congestive heart failure until 6 weeks was lower in the PTCA group (11.2% vs 17.6, P =.001). Mortality after 6 weeks (3.4% vs 5.6%, P =.07) and after 6 months (4.5% vs 7.1%, P =.06) tended to be lower in the PTCA group. CONCLUSION: Primary PTCA compared to thrombolytic therapy is associated with an accelerated myocardial reperfusion within 90 minutes after the start of reperfusion therapy. This early advantage in myocardial reperfusion is associated with an improved clinical outcome. PMID- 14564325 TI - Preprocedural white blood cell count and death after percutaneous coronary intervention. AB - BACKGROUND: Elevated inflammatory markers are associated with worse outcome after percutaneous coronary artery interventions (PCI). An elevation in the white blood cell (WBC) count is a nonspecific response to inflammation. We hypothesized that an elevated WBC count would be a predictor of death in patients undergoing PCI. METHODS: A total of 4450 patients undergoing percutaneous coronary artery intervention were divided into quintiles, based on their preprocedural WBC count (mean WBC count: quintile 1, 5.08 x 10(3)/muL; quintile 2, 6.58 x 10(3)/muL; quintile 3, 7.70 x 10(3)/muL; quintile 4, 9.14 x 10(3)/muL; and quintile 5, 13.4 x 10(3)/muL). Vital status was assessed through the use of the Social Security Death Index. RESULTS: There were a total of 504 deaths over a follow-up period of 48 months. The best survival was seen in quintile 2, with an increase in long term mortality rates seen with both a higher or a lower WBC count (P <.001). This J-shaped curve was preserved after multivariate adjustment, with the adjusted hazard ratio of mortality relative to quintile 2 being 1.95 (95% CI, 1.40 to 2.73) in quintile 1, 1.66 (95% CI, 1.18 to 2.33) in quintile 3, 2.31 (95% CI, 1.67 to 3.17) in quintile 4, and 2.42 (95% CI, 1.76 to 3.34) in quintile 5. CONCLUSIONS: A low or an elevated preprocedural WBC count in patients undergoing PCI is associated with an increased risk of long-term death. Our result provides further evidence to support the important role of inflammation in coronary artery disease. PMID- 14564326 TI - Safety of same-day-discharge radial percutaneous coronary intervention: a retrospective study. AB - BACKGROUND: The safety and feasibility of same-day discharge percutaneous coronary intervention (PCI) is still controversial. METHODS: Patients (n = 943) had same-day discharge radial PCI between April 1998 and March 2001 in our hospital. Patients were contacted and asked whether they had entry site complications or a repeat angiogram and/or PCI within 24 hours and 1 month after the procedure. RESULTS: At the time the study was conducted, 811 patients responded, 38 patients had died, and 94 were alive but refused to participate or it was impossible to contact them; 27 patients (2.8%) visited their doctor and/or the hospital within 24 hours after discharge because of entry site complications, and 38 patients (4.0%) visited within 1 month. However, none of the patients had major access site complications or needed to be admitted to the hospital. Within 24 hours from discharge 17 patients (2%) reported chest pain, and only 1 (0.1%) required a repeat angiogram, which did not show target vessel occlusion. During the first month, 94 patients (11.5%) reported chest pain, 11 (1.3%) underwent a repeat angiogram, out of which 4 had subacute vessel closure; 2 of the 132 patients that we could not contact had subacute stent thrombosis within 1 month and died. CONCLUSIONS: None of the patients having same-day discharge radial PCI had major access site complications. Six patients (0.6%) had subacute vessel closure, but none had this during the first 24 hours after discharge. Same-day discharge radial PCI in certain low-risk patients is a safe and feasible strategy. PMID- 14564327 TI - Randomized trial of a daily electronic home monitoring system in patients with advanced heart failure: the Weight Monitoring in Heart Failure (WHARF) trial. AB - BACKGROUND: Heart failure treatment guidelines emphasize daily weight monitoring for patients with heart failure, but data to support this practice are lacking. Using a technology-based heart failure monitoring system, we determined whether daily reporting of weight and symptoms in patients with advanced heart failure would reduce rehospitalization and mortality rates despite aggressive guideline driven heart failure care. METHODS: This was a randomized, controlled trial. Patients hospitalized with New York Heart Association class III or IV heart failure, with a left ventricular ejection fraction < or =35% were randomized to receive heart failure program care or heart failure program care plus the AlereNet system (Alere Medical, Reno, Nev) and followed-up for 6 months. The primary end point was 6-month hospital readmission rate. Secondary end points included mortality, heart failure hospitalization readmission rate, emergency room visitation rate, and quality of life. RESULTS: Two hundred eighty patients from 16 heart failure centers across the United States were randomized: 138 received the AlereNet system and 142 received standard care. Mean age was 59 +/- 15 years and 68% were male. The population had very advanced heart failure, New York Heart Association class III (75%) or IV (25%), as evidenced by serum norepinepherine levels, 6-minute walk distance and outcomes. No differences in hospitalization rates were observed. There was a 56.2% reduction in mortality (P <.003) for patients randomized to the AlereNet group. CONCLUSIONS: This is the largest multicenter, randomized trial of a technology-based daily weight and symptom-monitoring system for patients with advanced heart failure. Despite no difference in the primary end point of rehospitalization rates, mortality was significantly reduced for patients randomized to the AlereNet system without an increase in utilization, despite specialized and aggressive heart failure care in both groups. PMID- 14564328 TI - Effects of clopidogrel and aspirin combination versus aspirin alone on platelet aggregation and major receptor expression in patients with heart failure: the Plavix Use for Treatment Of Congestive Heart Failure (PLUTO-CHF) trial. AB - BACKGROUND: Persistent platelet activation may contribute to thrombotic events in patients with congestive heart failure (CHF). Chronic use of mild platelet inhibitors could therefore represent an independent avenue to improve morbidity, mortality, and quality of life in this expanding population. Although clopidogrel is widely used in patients with acute coronary syndromes and ischemic stroke, the ability of this novel ADP-receptor antagonist to inhibit platelet function in patients with CHF is unknown. We assessed antiplatelet properties of clopidogrel with aspirin (C+A) versus aspirin alone (A) in patients with CHF with heightened platelet activity. METHODS: Patients with left ventricular ejection fraction <40%, or CHF symptoms in the setting of preserved systolic function and New York Heart Association class II-IV were screened. Patients were considered to have platelet activation when 4 of the following 5 parameters were met: ADP-induced platelet aggregation >60%; collagen-induced aggregation >70%; whole blood aggregation >18 ohms; expression of GP IIb/IIIa >220 log MFI; and P-selectin cell positivity >8%. All patients were treated with 325 mg of acetylsalycilic acid (ASA) for at least 1 month. Patients receiving an antithrombotic agent other than ASA were excluded. Patients meeting clinical and laboratory criteria were randomly assigned to C+A (n=25), A (n=25) groups, or represent screen failures (n=38). Platelet studies (conventional and whole blood aggregometry, shear induced activation, expression of 10 major receptors and formation of platelet leukocyte microparticles) were performed at baseline and after 30 days of therapy. RESULTS: There were no deaths, hospitalizations, or serious adverse events. There were no changes in platelet parameters in the A group. In contrast, therapy with C+A resulted in a significant inhibition of platelet activity assessed by ADP-induced (P =.00001), and epinephrine-induced (P =.0016) aggregation, closure time (P =.04), expression of PECAM-1 (P =.009), GP Ib (P =.006), GP IIb/IIIa antigen (P =.0001), GP IIb/IIIa activity with PAC-1 (P =.0021), and CD151 (P =.0026) when compared with the A group. Therapy with C+A also resulted in the reduced formation of platelet-leukocyte microparticles (P =.021). Collagen-induced aggregation in plasma and in whole blood, expression of vitronectin receptor, P-selectin, CD63, CD107a, and CD107b did not differ among groups. CONCLUSIONS: Treatment with C+A for 1 month provides significantly greater inhibition of platelet activity than ASA alone in patients with CHF. Patients with CHF with heightened platelet activity represent a potential target population in which addition of clopidogrel may decrease mortality rates by reducing the incidence of thrombotic vascular events. PMID- 14564329 TI - Metoprolol CR/XL in postmyocardial infarction patients with chronic heart failure: experiences from MERIT-HF. AB - BACKGROUND: The benefit of beta-blockers post-myocardial infarction (MI) was established in the late 1970s. Major advances in the treatment of MI have since occurred. However, patients with chronic heart failure (CHF) were excluded from those trials. The purpose of this study was to assess the effect of beta-blockers in post-MI patients with CHF receiving contemporary management. METHODS: This was a prespecified subgroup analysis of a double-blind, randomized trial: the Metoprolol CR/XL Randomized Intervention Trial in Heart Failure (MERIT-HF). Patients with CHF in New York Heart Association class II to IV with an ejection fraction (EF) < or =0.40 and a history of being hospitalized for an acute MI (n = 1926) were randomized to metoprolol succinate controlled release/extended release (CR/XL) versus placebo. Mean EF was 0.28, and the mean follow-up was 1 year. RESULTS: Metoprolol CR/XL reduced total mortality by 40% (95% CI 0.20-0.55, P =.0004), and sudden death by 50% (95% CI 0.26-0.66, P =.0004). The combined end point of all-cause mortality/hospitalization for worsening CHF was reduced by 31% (95% CI 0.16-0.44, P <.0001), and cardiac death/nonfatal acute MI by 45% (95% CI 0.26-0.58, P <.0001). A post-hoc analysis showed that the outcome in patients with earlier revascularization (44%) and outcome in those with more severe CHF (20%) was similar to the entire post-MI population. CONCLUSIONS: In post-MI patients with symptomatic CHF, beta-blockade continues to exert a profound reduction in mortality and morbidity in the presence of contemporary management that includes early and late revascularization, angiotensin-converting enzyme inhibitors, aspirin, and statins. PMID- 14564330 TI - Combination of B-type natriuretic peptide and peak oxygen consumption improves risk stratification in outpatients with chronic heart failure. AB - BACKGROUND: Peak oxygen consumption is a cornerstone for prognostic determination in patients with congestive heart failure. The purpose of this study was to assess whether plasma B-type natriuretic peptide (BNP) provided any additional prognostic information. METHODS: Plasma concentrations of atrial natriuretic peptide, N terminal pro-atrial natriuretic peptide, BNP, endothelin-1, norepinephrine, and peak VO2 were measured in 250 consecutive outpatients with mild to moderate heart failure (96% in New York Heart Association [NYHA] class II or III) and left ventricular ejection fraction (LVEF) <45%. RESULTS: During a median follow-up of 584 days, 42 patients died (19 from sudden death) and 5 underwent urgent heart transplantation. Multivariate stepwise regression analysis showed that, among 13 variables including NYHA and LVEF, plasma BNP (chi2 = 11.9, P =.0001) was the strongest independent predictor of death or urgent transplantation, followed by serum sodium (chi2 = 8, P =.0046), resting heart rate (chi2 = 7.5, P =.0062), plasma endothelin-1 (chi2 = 7.2, P =.007), and peak VO2 (chi2 = 6.2, P =.012). Patients with plasma BNP above the upper quartile value (260 pg/mL) had a 1-year rate of death or urgent transplantation of 31%. The 1- and 2-year survival rates without urgent transplantation in patients with a peak VO2 < or =14 mL x kg(-1) x min(-1) were 71% and 59%, respectively, when plasma BNP was >137 pg/mL (median value), compared with 100% and 89%, respectively, when plasma BNP was < or =137 pg/mL (P =.008). Furthermore, plasma BNP was the only independent predictor of sudden death (chi2 = 19.9, P =.00001). CONCLUSIONS: Plasma BNP provides additive independent prognostic information compared to peak VO2 alone in outpatients with mild to moderate heart failure. PMID- 14564331 TI - Screening for left ventricular systolic dysfunction among patients with risk factors for heart failure. AB - BACKGROUND: The prevalence of left ventricular systolic dysfunction (LVSD) among individuals at risk for heart failure (HF) and the feasibility of screening have not been clearly defined. This study determined the prevalence of LVSD with the use of a limited screening echocardiogram among patients with risk factors for HF but no prior HF. METHODS: General medicine patients > or =60 years of age with hypertension, diabetes, coronary artery disease, or previous myocardial infarction (MI) but no history of HF or reduced left ventricular ejection fraction (LVEF) were eligible. Medical history and symptoms of breathlessness were determined by interview and chart review; consenting patients underwent electrocardiography and echocardiography. The outcome was LVEF < or =45%, based on visual estimation from the echocardiogram. RESULTS: Of the 482 patients who completed the study, only 1 patient could not have the LVEF visually estimated. A total of 7.9% of patients had LVEF < or =45%. The prevalence was 15.4% among those with a prior MI and 6.7% among those without prior MI. In multivariate analysis, prior MI (adjusted odds ratio, 2.75; 95% CI, 1.14 to 6.64) and probable or definite left ventricular hypertrophy by electrocardiography (adjusted odds ratio, 3.57; 95% CI, 1.22 to 10.48) were the strongest predictors of LVEF < or =45%. CONCLUSIONS: Screening for LVSD among high-risk patients is feasible and has substantial yield, even among patients without prior MI. In light of the low cost of screening and the available therapies to prevent progression of LVSD to overt HF, controlled clinical trials of screening high-risk subgroups appear to be justified. PMID- 14564332 TI - Transesophageal echocardiography before cardioversion of recurrent atrial fibrillation: does absence of previous atrial thrombi preclude the need of a repeat test? AB - BACKGROUND: Atrial fibrillation (AF) is a recurrent problem that frequently requires repeat cardioversion. Transesophageal echocardiography (TEE) is indicated before cardioversion in patients who are underanticoagulated (warfarin therapy <3 weeks or international normalized ratio [INR] <2.0). It remains uncertain if TEE should be repeated in underanticoagulated patients who had no atrial thrombi detected by previous TEE. Methods and results From January 1996 to June 2001, 76 patients (43 men, 33 women; mean age, 68.8 +/- 10.4 years) who were underanticoagulated and had no atrial thrombi in previous TEE underwent repeat TEE before cardioversion of recurrent AF. The duration of recurrent AF at the time of the second TEE was 5.1 +/- 9.3 months (1 day to 4 years). The underlying diseases included coronary artery disease (n = 30), hypertension (n = 22), valvular heart diseases (n = 8), dilated cardiomyopathy (n = 4), hypertrophic cardiomyopathy (n = 2), and others (n = 10). Eight (10.5%) patients (2 men, 6 women; mean age, 68.6 +/- 6.6 years) were found to have intra-atrial thrombi on the second TEE. Of these 8 patients, 3 had coronary artery disease, 1 had hypertension, 2 had dilated cardiomyopathy, 1 had hypertrophic cardiomyopathy, and 1 had AF of unknown cause. The duration of recurrent AF in patients with and without thrombi was not significantly different (3.6 +/- 4.7 versus 5.3 +/- 9.7 months, P =.22). Of the 8 patients with intra-atrial thrombi on the second TEE, 5 had been taking warfarin for 3 to 4 weeks but had subtherapeutic INR and 3 were taking aspirin only. Compared with patients without intra-atrial thrombi, patients with intra-atrial thrombi had lower ejection fraction (32.5% +/- 18.1% versus 49.9% +/- 14.1%, P =.015), slower left atrial appendage empty velocity (0.22 +/- 0.08 versus 0.41 +/- 0.17 m/s, P <.01), and higher prevalence of spontaneous echo contrast (87.5%) than in patients without intra-atrial thrombi (19.1%, P <.05) but similar left atrial size (49.5 +/- 5.3 versus 47.3 +/- 7.1 mm, P =.15). Cardioversion was cancelled in all patients with atrial thrombi. CONCLUSIONS: In underanticoagulated patients, repeat TEE is necessary before cardioversion of recurrent AF even if the previous TEE showed no atrial thrombi. PMID- 14564333 TI - Platelet activation after stenting with heparin-coated versus noncoated stents. AB - BACKGROUND: In animal models, heparin coating reduces platelet accumulation induced by coronary stenting. However, reduced platelet activation has never been demonstrated in humans. The purpose of the current study was to investigate the effect of heparin coating on platelet activation after coronary artery stenting. METHODS: In a prospective, randomized, pilot study of 50 consecutive elective patients, platelet activation was analyzed by measuring aggregation and surface receptor expression before and at 2 hours, 24 hours, 5 days and 30 days after implantation of either heparin-coated or noncoated stents. RESULTS: There was less platelet activation after implantation of a heparin-coated stent, as indicated by reduced expression at 24 hours of active glycoprotein (GP) IIb/IIIa (10.3 +/- 6.1 vs 6.7 +/- 2.1, P =.014), and total GP IIb/IIIa (382 +/- 101 vs 306 +/- 88, P =.01). A trend at 30 days poststenting of lower total (383 +/- 150 vs 296 +/- 86, P =.07) and active GP IIb/IIIa expression (10.3 +/- 6.9 vs 7.5 +/- 2.9, P =.15) was also observed with the heparin-coated stent). Aggregation and stimulated p-selectin did not differ between stent types. CONCLUSION: Use of a heparin-coated stent in this pilot study of elective patients was associated with primarily less early platelet expression of GP IIb/IIIa. These findings have implications on the risk of subacute thrombosis and deserve further investigation. PMID- 14564334 TI - Thermal hydrotherapy improves quality of life and hemodynamic function in patients with chronic heart failure. AB - BACKGROUND: Chronic heart failure is characterized by increased peripheral vascular resistance and reduced peripheral perfusion due to adrenergic and renin angiotensin activation and impaired endothelial function. Recent studies have shown that nonpharmacological peripheral vasodilation with thermal therapy by means of warm-water baths and sauna has beneficial effects in chronic heart failure. European hydrotherapy (according to Kneipp) additionally uses short cold water stimuli, which lead to prolonged vasodilation and adaptive responses. Studies on the efficacy of hydrotherapy in chronic heart failure are lacking. METHODS: We studied 15 patients (5 men, 10 women, mean (+/- SD) age 64.3 +/- 1.8 years) with mild chronic heart failure (NYHA functional class II to III, ejection fraction 30%-40%). Patients were randomly assigned to 6 weeks of intensive home based hydrotherapy or 6 weeks restriction in a crossover intervention trial. Quality of life and heart-failure--related symptoms were assessed by means of a validated questionnaire (PLC). Graded bicycle exercise test with incremental workloads (0, 50, 75, 100 watts) was performed at the end of each treatment period. The hydrotherapeutic program consisted of a structured combination of daily home-based external warm- and cold-water applications. RESULTS: Baseline characteristics were balanced between the groups. With hydrotherapy, a significant (P < or =.05) improvement in 3 of 6 dimensions of quality of life (mood, physical capacity, enjoyment) and a significant reduction in heart-failure related symptoms was found. Heart rates at rest and at 50-Watt workload were significantly reduced by hydrotherapy; blood pressure decreased nonsignificantly at rest and during exercise. The hydrotherapeutic treatment was well accepted and no relevant adverse effects were observed. CONCLUSIONS: A home-based hydrotherapeutic thermal treatment program improves quality of life, heart failure-related symptoms and heart rate response to exercise in patients with mild chronic heart failure. The results of this investigation suggest a beneficial adaptive response to repeated brief cold stimuli in addition to enhanced peripheral perfusion due to thermal hydrotherapy in patients with chronic heart failure. PMID- 14564335 TI - Current presentation and management of heart failure in cardiology and internal medicine hospital units: a tale of two worlds--the TEMISTOCLE study. AB - BACKGROUND: The purpose pf the current article is to describe the clinical profile, use of resources, management and outcome in a population of real-world inpatients with heart failure. METHODS AND RESULTS: With a prospective, cross sectional survey on acute hospital admissions, we evaluated the overall and provider-related differences in patient characteristics, diagnostic work-up, treatment and inhospital outcome of 2127 patients with heart failure admitted to 167 cardiology departments and 250 internal medicine departments between February 14 and 25, 2000. Patients admitted to cardiology units were younger (56.3% >70 years vs 76.2%, P <.0001), had more severe symptoms (NYHA IV 35% vs 29%, P =.00014), and more often underwent evaluation of ventricular function (89.3% vs 54.8%, P <.0001) and coronary angiography (7.5% vs 0.9%, P <.0001) than those admitted to medical units. Moreover, they were more often prescribed beta blockers (17.8% vs 8.7%, P <.0001). However, prescription of angiotensin converting enzyme inhibitors and angiotensin-receptor blockers (78.7% vs 81.5%, P = not significant [NS]) and inhospital mortality (5.2% vs 5.9%, P = NS) were similar. A 6-month follow-up visit was performed in 56.4% of cases (68.2% of cardiology vs 49.4% of medicine patients, P <.0001); 6-month readmission (43.7% vs 45.4%, P = NS) and mortality (13.9% vs 16.7%, P = NS) rates were similar. CONCLUSIONS: Patients with heart failure admitted to cardiology and internal medicine units represent 2 clearly different populations. In both groups, diagnostic procedures and evidence-based treatments, such as beta-blockers, appeared to be underused, and there was a lack of structured follow-up, as well as a poor 6-month prognosis. PMID- 14564336 TI - Does the type of prosthesis influence early left ventricular mass regression after aortic valve replacement? Assessment with magnetic resonance imaging. AB - BACKGROUND: Debate exists regarding selection of the prosthesis type most likely to maximize early left ventricular (LV) mass regression after aortic valve replacement (AVR) for stenotic valvular disease. The aim of this study was to compare the degree of LV mass regression measured by MRI 6 months after prospectively randomized valve implantation for two biological prostheses, stented and stentless, and for two mechanical valves, tilting disc and bileaflet. METHODS: Thirty-nine consecutive patients with predominant aortic stenosis accepted for elective AVR were studied. Twenty patients requiring a tissue prosthesis were randomly assigned to receive either a Freestyle or Mosaic valve. The remaining 19 patients in whom mechanical prosthesis was indicated were randomly assigned to receive either an Ultracor or an ATS valve. RESULTS: There was no difference in valve size implanted between the compared groups. LV mass measurements were performed with MRI (1.5-T Vision, Siemens, Germany) immediately before and 6 months after surgery. All valve types produced significant postoperative reduction in LV mass compared with preoperative values (P <.01). Percent change in LV mass regression was similar between the two porcine valve types, Mosaic (24.4% +/- 11.1%) and Freestyle (21.1% +/- 16.7%), and between the two mechanical valve designs, Ultracor (19.3% +/- 9.5%) and ATS (26.3% +/- 10.8%), respectively. CONCLUSIONS: Significant LV remodeling occurs early after AVR for aortic stenosis. The degree of regression in LV mass is independent of prosthesis type implanted. PMID- 14564338 TI - Hiccup as an electrocardiographic artifact simulating arrhythmias. PMID- 14564337 TI - Effect of transdermal nitroglycerin on inflammatory mediators in patients with peripheral atherosclerotic vascular disease. AB - OBJECTIVE: To demonstrate that nitroglycerin improves biological markers of arterial inflammation in patients with peripheral vascular disease. BACKGROUND: Atherosclerosis is an inflammatory disease in which there is an increase in active inflammation markers such as C-reactive protein and other factors released by endothelial cells. Nitroglycerin acts by a chemical liberation of nitric oxide. We have previously published the results from several controlled clinical trials confirming an anti-inflammatory action of nitroglycerin. METHODS: Forty patients with peripheral vascular disease entered a randomized, double-blind, placebo-controlled pilot study for 6 weeks. Twenty-one patients were treated with continuous application of a transdermal nitroglycerin patch (15 mg/24 hours) on the anterior face of the thigh. Venous blood samples were obtained before treatment and 2 and 6 weeks after. We measured plasma levels of C-reactive protein, cGMP (also intraplatelet cGMP), E-selectin, ICAM, VCAM-1, IL-6, and nitrites/nitrates. RESULTS: No biological parameter was modified in the placebo group. On the contrary, nitroglycerin significantly reduced plasma levels of C reactive protein and sE-selectin and increased the levels of intraplatelet cGMP. CONCLUSIONS: The results of this preliminary study show that nitroglycerin has an anti-inflammatory action in patients with peripheral vascular disease. This may provide a new therapeutic approach to understanding the efficacy of nitrovasodilators in the improvement of atherosclerotic syndromes. PMID- 14564340 TI - Obstructive sleep apnea. AB - Obstructive sleep apnea is an increasingly well-recognized disease characterized by periodic collapse of the upper airway during sleep. This leads to either complete or partial obstruction of the airway, resulting in apneas, hypopneas, or both. This disorder causes daytime somnolence, neurocognitive defects, and depression. It affects almost every system in the body, resulting in an increased incidence of hypertension, cardiovascular disease, stroke, pulmonary hypertension, cardiac arrhythmias, and altered immune function. It also increases the risk of having an accident, presumably as a result of associated somnolence. The gold standard for the diagnosis of sleep apnea is an overnight polysomnogram. Split-night studies are becoming increasingly common and allow for quicker implementation of therapy at a reduced cost. Treatment options for sleep apnea include weight loss, positional therapy, oral devices, continuous positive airway pressure (CPAP), and upper airway surgery. CPAP is the most efficacious and widely used therapy. Its complications include nasal congestion or dryness, mask discomfort, and claustrophobia. Heated humidifiers, newer types of masks, and nasal steroids have improved tolerance of this therapy. Bilevel positive-pressure therapy can be considered for patients who find it difficult to exhale against the consistently increased pressure of CPAP. The disease requires aggressive treatment to improve quality of life and prevent its complications. PMID- 14564341 TI - Biology of common beta receptor-signaling cytokines: IL-3, IL-5, and GM-CSF. AB - IL-3, IL-5, and GM-CSF are related hematopoietic cytoines that are important for allergic inflammation. The receptors for human IL-5, IL-3, and GM-CSF are members of the hematopoietin receptor superfamily and are comprised of a cytokine specific alpha chain and the common beta chain that is shared among these cytokines for signaling. Each of these cytokines contributes to the differentiation and function of leukocyte subpopulations and have clinical importance in protective immunity and in the pathophysiology of a spectrum of immunologic diseases that are as diverse as allergy and asthma, pulmonary alveolar proteinosis, neurodegenerative diseases, and malignancies. Delineating the biology of these cytokines is enabling the development of new strategies for diagnosing and treating these diseases and modulating immune responses. PMID- 14564342 TI - Viral infections in atopic dermatitis: pathogenic aspects and clinical management. AB - A number of different widespread and disseminated viral infections can occur in patients with atopic dermatitis. Eczema molluscatum is troublesome but not dangerous. Although eczema vaccinatum is rare, it is life-threatening and of increased concern as smallpox vaccinations are reintroduced as a response to possible bioterrorism. There is little information on the course of smallpox itself in atopic dermatitis. Eczema herpeticum is the most common member of this group; recent advances in understanding its pathogenesis might contribute to a more successful management of this serious complication. PMID- 14564343 TI - Combating high-priority biological agents: What to do with drug-allergic patients and those for whom vaccination is contraindicated? AB - The threat of bioterrorism continues to be a very real one. Regularly, there are news stories on bioterrorism-related topics: What biologic weapons will our enemies likely use to attack the United States? How prepared is our country to successfully counter such attacks? Although these critical questions are being addressed by the leaders of our country, allergists-immunologists, too, will have to grapple with difficult questions during these uncertain and frightening times. We care for a special group of patients with various allergic and immunologic disorders. Some of our patients have immunodeficiency disorders that might preclude them from receiving life-saving vaccines. Our patients with drug allergies are fearful that should they become infected with a biologic agent, they will not be able to receive appropriate treatment. In this article we focus on the various vaccine-related and antibiotic-related adverse effects that the allergist-immunologist might see during treatment of infections caused by Category A agents. Where possible, potential management approaches are outlined. PMID- 14564344 TI - Risks of smallpox vaccination: 200 years after Jenner. PMID- 14564345 TI - Innate immune activation as a broad-spectrum biodefense strategy: prospects and research challenges. AB - Biodefense strategies require protection against a broad and largely unforeseen spectrum of pathogens--the forte of innate immune system defenses--that have evolved over millennia to function within moments of encountering either ancient or newly emerging pathogens. Although constitutive, the innate immune system is activated by the presence of microbes or their products, providing a rationale for a potential biodefense strategy. Both prophylactic and postexposure strategies involving innate immune stimulation have been shown to be plausible to prevent or ameliorate infections in animal models. Innate immune-activating compounds based on conserved microbial components recognized by toll-like molecules and other receptors could be synthesized and delivered like drugs by using an entirely different strategy from conventional vaccination. However, important theoretic and practical questions emerge about developing and deploying innate immune protective strategies for biodefense. This rostrum discusses prospects and problems in the overall approach itself. Important topics include microbe-specific issues about innate immune system effectiveness against highly virulent pathogens and general questions, such as whether innate immune responses will be safe and effective if used in a diverse human population of different age groups and with different genetic makeups. PMID- 14564346 TI - Elevated substance P levels in nasal lavage fluids from patients with chronic nonproductive cough and increased cough sensitivity to inhaled capsaicin. AB - BACKGROUND: The exact mechanism of a chronic nonproductive cough is sometimes unclear when patients who are without symptoms or signs indicating the major causes of chronic cough remain undiagnosed. OBJECTIVE: We hypothesized that some neurochemical alterations in the sensory nerves in the cough reflex may occur in the upper airway of chronic nonproductive cough patients. METHODS: We took nasal lavage fluid (NLF) specimens from 38 patients with a chronic nonproductive cough as the sole presenting symptom. All 38 had normal chest radiography, spirometry, and bronchial responsiveness. We likewise took NLF specimens from 14 healthy control subjects. We used a capsaicin cough provocation test to determine cough sensitivity and considered the value of C5 (the lowest capsaicin concentration inducing 5 consecutive coughs) as an index of cough sensitivity. We measured levels of substance P of NLF specimens by using ELISA. In addition, we evaluated the clinical response of each patient after subsequent therapeutic trials with an antihistamine and decongestant for 2 weeks. RESULTS: By using capsaicin cough sensitivity as the basis for grouping the study subjects, we divided the patients into 2 groups: an increased cough sensitivity group (ICS, C5 <32 mumol/L) and a normal cough sensitivity (NCS) group. Patients with ICS showed an elevated SP concentration in NLF (median value, 408 pg/mL) compared with that of the NCS group (237 pg/mL) and the control subjects (138 pg/mL) (P <.01). The median value of the percentage of remnant cough after therapeutic trial compared with the cough status before treatment was significantly higher in the ICS subgroup (70%) than that of NCS (25%) (P <.05). CONCLUSIONS: Elevated substance P contents in NLF specimens were associated with ICS in patients with chronic nonproductive cough, suggesting a neurochemical abnormality in the upper airway. PMID- 14564347 TI - Urinary eosinophilic protein X, atopy, and symptoms suggestive of allergic disease at 3 years of age. AB - BACKGROUND: Urinary eosinophilic protein X (U-EPX) measurement is easy to perform in children. However, its use for prediction, diagnosis, and monitoring of asthma and atopy is unclear. OBJECTIVE: We sought to investigate the relationship between U-EPX and clinical phenotypes suggestive of allergic diseases. METHODS: U EPX measurement (RIA), respiratory questionnaires, and skin testing were completed at age 3 years in 903 children followed prospectively from birth. Specific airway resistance was measured in 503 currently asymptomatic children by using whole-body plethysmography during tidal breathing. RESULTS: Nonatopic children with wheezing or eczema had slightly increased U-EPX levels compared with nonatopic asymptomatic children. U-EPX levels (geometric mean EPX/creatinine ratio) were as follows: nonatopic asymptomatic children (n = 313), 61.3 microg/mmol (95% CI, 56.4-66.6 microg/mmol); nonatopic children with wheezing (n = 148), 71.2 microg/mmol (95% CI, 63.2-80.1 microg/mmol); nonatopic children with eczema (n = 90), 65.7 microg/mmol (95% CI, 56.7-76.2 microg/mmol); and nonatopic children with wheezing and eczema (n= 86), 79.7 microg/mmol (95% CI, 67.4-94.3 microg/mmol). Children who had persistent atopy early in life had significantly higher U-EPX levels at age 3 years (nonatopic at 1 and 3 years [n = 263], 63.4 microg/mmol [95% CI, 58.4-69.0 microg/mmol]; atopic at 1 but not 3 years [n = 24], 65.1 microg/mmol [95% CI, 43.8-96.7 microg/mmol]; nonatopic at 1 year and atopic at 3 years [n = 62], 90.0 microg/mmol [95% CI, 74.6-108.4 microg/mmol]; atopic at 1 and 3 years [n = 35], 111.5 microg/mmol [95% CI, 89.2-139.3 microg/mmol]; P <.002). Atopy alone and with wheezing, eczema, or both was associated with significantly increased U-EPX levels (P <.0001). Wheezing appeared to be associated with higher U-EPX levels compared with eczema in both atopic and nonatopic children. The highest U-EPX level was found in atopic children with a history of wheezing and eczema (P <.0001). There was no relationship between U-EPX level and lung function. CONCLUSION: U-EPX level reflects the presence of atopy and associated symptoms and might be useful for monitoring the progression of allergic disease. PMID- 14564348 TI - Increased prostaglandin E2 concentrations and cyclooxygenase-2 expression in asthmatic subjects with sputum eosinophilia. AB - BACKGROUND: Prostaglandin E2 (PGE2) is known to be produced within human airways, but it is not clear whether in airway diseases it can play a deleterious or a beneficial role. Recently it has been reported that PGE2 can enhance eosinophil survival in vitro. OBJECTIVE: To evaluate whether the concentrations of PGE2 in asthmatic airways correlate with the number of eosinophils and can be responsible for eosinophil-enhanced survival and to identify the cyclooxygenase isoform contributing to the synthesis of PGE2 by cells present in asthmatic airways. METHODS: Reversed-phase high-performance liquid chromatography and/or specific radioimmunoassay was used to measure PGE2 concentrations in induced sputum supernatants from 14 control and 30 asthmatic subjects. Correlations between concentrations of PGE2 and the number of eosinophils in induced sputum were evaluated. Expression of cyclooxygenase-2 (COX-2) in induced sputum cells was determined by immunocytochemistry, and the effect of COX-2 inhibition on PGE2 production was evaluated with the use of radiolabeled arachidonic acid. The effects on eosinophil apoptosis by PGE2 or induced sputum supernatants were studied by using peripheral blood eosinophils obtained by negative immunomagnetic selection. RESULTS: PGE2 concentrations resulted in elevated samples from asthmatic subjects and directly correlated with the percentage of eosinophils and the concentrations of eosinophilic cationic protein. Immunostaining for COX-2 showed enhanced expression in macrophages of asthmatic subjects when compared with control subjects, and the use of a specific COX-2 inhibitor provided evidence that PGE2 synthesis was the result of COX-2 enzymatic activity in asthma induced sputum cells. Supernatant from induced sputum of asthmatic subjects with high eosinophil counts caused a decreased apoptosis of peripheral blood eosinophils when compared with control subjects, and immunoprecipitation of PGE2 significantly reverted this phenomenon, suggesting that PGE2 was present in biologically relevant concentrations in induced sputum. CONCLUSIONS: The results obtained suggest that COX-2 expression in alveolar macrophages from asthmatic subjects may contribute to enhanced eosinophil survival through an increased PGE2 production. PMID- 14564349 TI - Association of a disintegrin and metalloprotease 33 (ADAM33) gene with asthma in ethnically diverse populations. AB - BACKGROUND: Asthma is a complex genetic disease characterized by reversible intermittent airway obstruction and respiratory symptoms primarily caused by acute and chronic bronchial inflammation. Recently, a gene potentially involved in airway remodeling, a disintegrin and metalloprotease 33 (ADAM33), was implicated in asthma susceptibility. OBJECTIVE: We sought to determine whether polymorphisms in ADAM33 are associated with asthma or closely related phenotypes in 4 different asthma populations. METHODS: Eight single nucleotide polymorphisms (SNPs) were evaluated in the 3' portion of ADAM33 in 4 unique asthma populations (African American, US white, US Hispanic, and Dutch white). These SNPs were previously reported to be associated with asthma in white populations from the United States and United Kingdom. RESULTS: Significant associations were observed with at least one SNP and asthma in each population (P =.0009-.04). Related phenotypes that included total serum IgE levels and skin test responsiveness were also associated (P =.003-.05). However, no single SNP was associated across all populations. Additionally, haplotype analysis revealed that no single haplotype accounted for asthma susceptibility risk, although potential risk haplotypes existed within some of the populations. CONCLUSION: Replication of the original ADAM33 findings in these 4 additional asthma populations suggests that this gene (and perhaps others that interact with it) is important in the development and pathogenesis of asthma. PMID- 14564350 TI - TGF-beta in human milk is associated with wheeze in infancy. AB - BACKGROUND: Cytokines secreted in human milk might play important roles in newborn health and in the development of infant immune responses. We investigated the relationship of the concentration and dose of cytokines in human milk to infant wheeze at 1 year of age. OBJECTIVE: Our objective was to test whether the cytokines in milk could account for some of the apparent protective effect of breast-feeding against wheeze in the first year of life. METHODS: Data on breast feeding and infant wheeze were collected prospectively from birth to 1 year from 243 mothers participating in the Infant Immune Study in Tucson, Arizona. Breast milk samples obtained at a mean age of 11 days postpartum were assayed by means of ELISA for concentrations of TGF-beta1, IL-10, TNF-alpha, and the soluble form of CD14. The dose of each cytokine was assessed for a relationship with wheeze in bivariate and logistic regression analyses. RESULTS: Increasing duration of breast-feeding was significantly associated with a decreased prevalence of wheeze (P =.039). There was wide variability in levels of each cytokine in milk, as well as variability between women in the amount of each cytokine produced. There was a significant inverse association between the dose of TGF-beta1 received through milk with the percentage of wheeze (P =.017), and the relationship was linear (P =.006). None of the other cytokines showed a linear relationship with wheeze. In multivariate analyses the risk of wheeze was significantly decreased (odds ratio, 0.22; 95% CI 0.05-0.89; P =.034) with increasing TGF-beta1 dose (long breast feeding and medium-high TGF-beta1 level compared with short breast-feeding and low TGF-beta. CONCLUSION: This analysis shows that the dose of TGF-beta1 received from milk has a significant relationship with infant wheeze, which might account for at least some of the protective effect of breast-feeding against wheeze. PMID- 14564351 TI - The development of asthma in children infected with Chlamydia pneumoniae is dependent on the modifying effect of mannose-binding lectin. AB - BACKGROUND: Although several studies found associations between infection with Chlamydia pneumoniae and asthma, these were mainly restricted to the exacerbation of the symptoms in adults with known asthma. Data about the role of C pneumoniae in the initiation and development of asthma in children are controversial. OBJECTIVE: We investigated the role of C pneumoniae infection in 139 children with asthma, comparing them with 174 healthy control subjects. Furthermore, we studied the modifying effect of mannose-binding lectin (MBL) variant alleles on the susceptibility to asthma in children infected with C pneumoniae. METHODS: C pneumoniae-specific antibodies were measured by means of ELISA, and MBL genotypes were determined by means of PCR-RFLP. RESULTS: There were no significant differences in the percentage of children with positive results for C pneumoniae specific antibodies between patients and control subjects. Among asthmatic children carrying variant MBL alleles, there were significantly more patients with positive results for C pneumoniae-specific IgG than among control children with variant MBL genotypes (63.7% vs 40.7% of asthmatic vs control children, respectively; odds ratio adjusted for age and sex, 2.21; 95% CI, 1.10-4.41; P =.02). Infected children with variant MBL alleles were found to have a higher risk of asthma development than infected children with normal MBL genotype. This risk was especially high in children with chronic or recurrent infection (positive results for both IgA and IgG; adjusted odds ratio, 5.38; 95% CI, 1.75 14.36; P =.01), but no increased risk was seen in children with current C pneumoniae infection (positive results for IgM). CONCLUSION: This study indicates the important role of variant MBL alleles in the susceptibility to asthma in children infected with C pneumoniae. PMID- 14564352 TI - Association study of the IL13 variant Arg110Gln in atopic diseases and juvenile idiopathic arthritis. AB - BACKGROUND: It has previously been shown that various inflammatory diseases, such as diabetes mellitus, bronchial asthma, chronic inflammatory bowel diseases, and rheumatoid arthritis, are in some circumstances genetically linked to the same chromosomal regions. Consequently, common genes underlying the pathogenetics of these diseases have been proposed. Chronic inflammatory disorders can be subdivided by their predominant immune response, either TH1 or TH2. For example, juvenile idiopathic arthritis (JIA) is a TH1 disease, and bronchial asthma is a TH2 disease. OBJECTIVES: The present study investigated the polymorphism Arg110Gln within the IL13 gene, a strong TH2 cytokine. We attempted to determine whether it is associated with these 2 diseases and whether this would reflect the TH1/TH2 paradigm. METHODS: Arg110Gln was typed in 4 different populations: asthmatic children, atopic children, children with JIA, and a control population. Statistical analysis was performed by using logistic and linear regression analysis of serum IgE levels and the Armitage trend test. RESULTS: The variant Gln110 was shown to be associated with increased total serum IgE levels in our atopic population (P =.006) and was weakly associated with bronchial asthma (P =.04). There was no association of the variant with JIA when compared with the control population. However, the variant Gln110 was significantly less frequent in children with JIA compared with its presence in children with bronchial asthma (P =.007). CONCLUSION: This is the first study to compare the same gene variant in TH1 and TH2 chronic inflammatory diseases. The results suggest that the same gene variant might protect from one disease and make an individual susceptible to the other. PMID- 14564354 TI - Developmental cytokine response profiles and the clinical and immunologic expression of atopy during the first year of life. AB - BACKGROUND: Allergic diseases have been linked to abnormal patterns of immune development, and this has stimulated efforts to define the precise patterns of cytokine dysregulation that are associated with specific atopic phenotypes. OBJECTIVE: Cytokine-response profiles were prospectively analyzed over the first year of life and compared with the clinical and immunologic expressions of atopy. METHODS: Umbilical cord and 1-year PBMCs were obtained from 285 subjects from allergic families. PHA-stimulated cytokine-response profiles (IL-5, IL-10, IL-13, and IFN-gamma) were compared with blood eosinophil counts and total and specific IgE levels (dust mites, cat, egg, Alternaria species, peanut, milk, and dog) at age 1 year and at the development of atopic dermatitis and food allergy. RESULTS: For the cohort as a whole, cytokine responses did not evolve according to a strict TH1 or TH2 polarization pattern. PHA-stimulated cord blood cells secreted low levels of IL-5 (2.1 pg/mL), moderate levels of IFN-gamma (57.4 pg/mL), and greater amounts of IL-13 (281.8 pg/mL). From birth to 1 year, IL-5 responses dramatically increased, whereas IL-13 and IFN-gamma responses significantly decreased. Reduced cord blood secretion of IL-10 and IFN-gamma was associated with subsequent sensitization to egg. In addition, there was evidence of TH2 polarization (increased IL-5 and IL-13 levels) associated with blood eosinophilia and increased total IgE levels by age 1 year. CONCLUSION: These findings demonstrate that cytokine responses change markedly during the first year of life and provide further evidence of a close relationship between TH2 skewing of immune responses and the incidence of atopic manifestations in children. PMID- 14564356 TI - Four-year incidence of allergic sensitization among schoolchildren in a community where allergy to cat and dog dominates sensitization: report from the Obstructive Lung Disease in Northern Sweden Study Group. AB - BACKGROUND: Exposure to high levels of cat allergen might prevent sensitization. OBJECTIVE: We sought to measure the incidence of allergic sensitization among schoolchildren living in a dust mite- and cockroach-free environment and the associated risk factors. METHODS: In 1996, a longitudinal cohort was established in northern Sweden, including 2454 children aged 7 to 8 years. Children were skin tested, and the testing was repeated 4 years later. Questionnaires were completed yearly. Participation was 88% both in 1996 and 2000. RESULTS: The prevalence of positive skin test results increased from 20.6% at age 7 and 8 years to 30.4% at age 11 and 12 years, a cumulative incidence of 13.8%, and was significantly higher among boys. The incidence was highest for cat (6.0%), timothy grass (5.9%), dog (4.9%), and birch (3.6%). A family history of allergy was the major risk factor for both a positive skin test response at age 7 and 8 years (odds ratio [OR], 1.69; 95% CI, 1.36-2.10) and for development of a positive skin test response over the next 4 years (OR, 1.67; 95% CI, 1.23-2.28). A significant inverse association between cat and dog ownership and the prevalence of type 1 allergy was found, particularly for those children who had lived with a cat both before age 7 and 8 years and during the next 4 years (OR, 0.44; 95% CI, 0.31 0.61). A similar pattern, although not significant, was found for incident cases. CONCLUSION: The high incidence of type 1 allergy at this age was similar to reports from communities with mite and cockroach allergen. Despite cat and dog being the most common allergens of sensitization, keeping these animals at home was not associated with an increased risk for sensitization. PMID- 14564359 TI - Endogenous nitric oxide downregulates the Bcl-2 expression of eosinophils through mitogen-activated protein kinase in bronchial asthma. AB - BACKGROUND: Eosinophil apoptosis might play a crucial role in the maintenance of persistent airway inflammation in asthma. Nitric oxide (NO) synthase activity is upregulated in eosinophils of asthmatic patients. Mitogen-activated protein kinase (MAPK) is implicated in regulating eosinophil apoptosis by modulating Bcl 2 expression. NO-induced cell apoptosis is associated with an inhibition of Bcl-2 expression. OBJECTIVE: We sought to study whether NO might induce eosinophil apoptosis through extracellular signal-regulated protein kinase (ERK) or p38 MAPK pathways and Bcl-2 expression. METHODS: Eosinophils were freshly isolated from peripheral blood of 16 asthmatic patients and 12 healthy subjects and then cultured with or without the NO synthase inhibitor N-nitro-l-arginine methyl ester (L-NAME) at 1 and 10 mmol/L for 16 hours. The expression of Bcl-2 and induced NO synthase on eosinophils was analyzed by means of flow cytometry. Apoptosis was assessed by means of propidium iodide and DNA ladder. The activity of ERK and p38 MAPK was determined by means of Western blotting. RESULTS: The induced NO synthase immunoreactivities and the spontaneous release of nitrite from the eosinophils of asthmatic patients were higher compared with those of healthy subjects. Eosinophils of asthmatic patients were found to express more highly constitutive Bcl-2 than those of healthy subjects. After incubation for 16 hours, the expression of Bcl-2 on eosinophils from patients with asthma was significantly enhanced by L-NAME. The percentage of apoptosis was decreased by the addition of 1 mmol/L L-NAME in patients with asthma. The activity of p38 MAPK and ERK in eosinophils from patients with asthma was enhanced in the presence of L-NAME. An inhibition of MAPK reduced the Bcl-2 expression and increased eosinophil apoptosis in patients with asthma. CONCLUSION: We concluded that inhibition of endogenous NO might increase the expression of Bcl-2 in eosinophils from patients with asthma through the signaling pathway of ERK or p38 MAPK, which in turn decrease the apoptosis. PMID- 14564360 TI - 5-oxo-6,8,11,14-eicosatetraenoic acid induces the infiltration of granulocytes into human skin. AB - BACKGROUND: 5-Oxo-6,8,11,14-eicosatetraenoic acid (5-oxo-ETE) is an arachidonic acid metabolite with potent in vitro chemoattractant effects on eosinophils and neutrophils. It has also been shown to induce pulmonary eosinophilia in Brown Norway rats, but it is not known whether it is active in human beings in vivo. OBJECTIVE: To determine whether 5-oxo-ETE can induce cellular infiltration in patients with atopic asthma and nonatopic control subjects after intradermal administration. METHODS: 5-Oxo-ETE was administered intradermally to 11 patients with atopic asthma and 10 nonatopic control subjects. Skin biopsy specimens were taken 6 or 24 hours later and examined by immunocytochemistry for cells expressing specific markers for eosinophils (major basic protein), neutrophils (elastase), macrophages (CD68), lymphocytes (CD3), and mast cells (tryptase). RESULTS: 5-Oxo-ETE (1.5 and 5 microg) elicited the infiltration of both eosinophils and neutrophils into the skin in both control and atopic asthmatic subjects. Increased numbers of eosinophils were observed at 6 and 24 hours after injection, whereas significantly elevated neutrophil numbers were present only after 24 hours. Eosinophils were >3 times higher in patients with atopic asthma compared with control subjects after injection of the highest dose of 5-oxo-ETE. Macrophage numbers were also elevated, but only at the highest dose of 5-oxo-ETE. No effects were observed on the numbers of either lymphocytes or mast cells. CONCLUSIONS: 5-Oxo-ETE elicits the infiltration of eosinophils and neutrophils into the skin of human beings in vivo after intradermal administration. Asthmatic subjects are more responsive to this substance than nonallergic control subjects. These results suggest that 5-oxo-ETE may be an important mediator of inflammation. PMID- 14564357 TI - Hyper-IgM syndrome with putative dominant negative mutation in activation-induced cytidine deaminase. AB - BACKGROUND: Hyper-IgM immunodeficiency is an immunologic disorder characterized by normal or increased serum IgM levels and reduced serum IgG and IgA levels caused by the disruption of Ig class switching in B cells. The gene encoding activation-induced cytidine deaminase (AID) is responsible for the autosomal recessive form of hyper-IgM syndrome. OBJECTIVE: To investigate the relationship between the AID gene mutation and the clinical phenotype, we analyzed the AID gene in a female Japanese patient with the autosomal recessive form of hyper-IgM syndrome. METHODS: Genomic DNA and cDNA were extracted from neutrophils and analyzed by means of PCR. The AID gene was expressed as a glutathione S transferase fusion protein. RT-PCR was performed after stimulation of the patient's PBMCs with phorbol myristate acetate and TGF-beta. RESULTS: Despite significantly low serum IgG levels, our patient had not shown a predisposition to any severe infections, even without Ig replacement therapy. We identified a point mutation resulting in the stop codon in exon 5 of the AID gene (R190X) in the patient. No other mutations of the AID gene were detected in the patient. The same mutation was not detected in other members of her family. The mutant allele fused with the glutathione S-transferase protein was expressed stably at the same level as the normal allele. The AID gene expression in the patient was induced by phorbol myristate acetate and TGF-beta. CONCLUSION: The mutation of the AID gene is assumed to be of the dominant negative form. This is the first report of a dominant negative form of the mutation in vivo. PMID- 14564361 TI - Lipid-transfer protein is the major maize allergen maintaining IgE-binding activity after cooking at 100 degrees C, as demonstrated in anaphylactic patients and patients with positive double-blind, placebo-controlled food challenge results. AB - BACKGROUND: In a previous study a 9-kd lipid-transfer protein (LTP) was identified as the major allergen of raw maize in a population of 22 anaphylactic patients. However, the stability of this protein in cooked maize is unknown. OBJECTIVE: We investigated the allergenicity of 5 maize hybrids and its modification after different thermal treatments by using sera from anaphylactic patients and patients with positive double-blind, placebo-controlled food challenges. METHODS: Five maize hybrids were extracted by using different methods, obtaining the water-soluble, zein, total zein, glutelin, and total protein fractions. The IgE-binding capacity of the different extracts, both raw and after thermal treatment, was investigated by means of SDS-PAGE immunoblotting. A 9-kd heat-stable allergen was purified by means of HPLC and sequenced. Changes in its secondary structure during and after heating from 25 degrees C to 100 degrees C were monitored by means of circular dichroism. RESULTS: All raw maize hybrids showed similar protein and IgE-binding profiles. The SDS-PAGE of all the heat-treated hybrids demonstrated a decreased number of stained bands in respect to the raw samples. The IgE immunoblotting demonstrated that the major allergen of the water-soluble, total zein, total protein, and glutelin fractions was a 9-kd protein identified by means of amino acid sequence as an LTP and a sub-tilisin-chymotrypsin inhibitor (in total zein fraction). The IgE-binding capacity of this 9-kd protein remained unchanged after thermal treatments, even though circular dichroism demonstrated an altered secondary structure. CONCLUSIONS: Maize LTP maintains its IgE-binding capacity after heat treatment, thus being the most eligible candidate for a causative role in severe anaphylactic reactions to both raw and cooked maize. PMID- 14564362 TI - A survey on the management of pollen-food allergy syndrome in allergy practices. AB - BACKGROUND: There is no consensus on the diagnosis and therapy of oral allergy syndrome (OAS; also known as pollen-food allergy syndrome), a disorder caused by IgE antibody-mediated reactions to homologous proteins in pollens and fruits and vegetables. OBJECTIVE: We sought to determine how practicing allergists define and treat OAS. METHODS: A questionnaire was mailed to 226 randomly selected US allergists from the American Academy of Allergy, Asthma and Immunology directory. RESULTS: One hundred twenty-two (54%) returned surveys were analyzed. Median estimates of the prevalence of OAS among the patients with pollen allergy were 5% among children and 8% among adults. Twenty percent of allergists reported that some patients progressed to systemic symptoms. Fifty-three percent of allergists recommended complete avoidance of causal foods to all patients, whereas 9% did not advocate any restrictions. Thirty percent never prescribed epinephrine for OAS, 3% always did, and the remainder did so on the basis of symptoms. When presented with clinical cases, 20% diagnosed systemic reactions to peach as OAS, 13% believed peanut could cause OAS, and 25% did not prescribe epinephrine for peanut allergy manifested by oral symptoms. CONCLUSION: Allergists' estimates of the prevalence of OAS in patients with pollen allergy (5%-8%) are lower than the prevalence reported (approximately 50%) in the published studies of these patients, perhaps reflecting a low index of suspicion, underdiagnosis, or both. The wide range of responses regarding diagnosis and management indicates the need for a better definition for the disorder and standard therapeutic guidelines. Discrepancies might be related to the term OAS, and therefore use of the more specific term "pollen-food allergy syndrome" is suggested. PMID- 14564365 TI - Pollen and eosinophilic esophagitis. PMID- 14564363 TI - Clinically relevant peach allergy is related to peach lipid transfer protein, Pru p 3, in the Spanish population. AB - BACKGROUND: Sensitization to peach and related Rosaceae fruits without clinical expression is commonly observed as the result of the extensive cross-reactivity of IgE antibodies directed toward lipid transfer proteins (LTPs), Bet v 1 homologues, profilins, and carbohydrate determinants. OBJECTIVE: We aimed to study whether there are any clinical or immunologic differences between patients allergic to peach and those who have a current clinically irrelevant sensitization to this fruit. METHODS: One hundred subjects with adverse reactions to peach were evaluated by medical history, skin prick tests with fresh peach and purified peach LTP (Pru p 3), and specific IgE determinations to peach, rBet v 1, and rBet v 2 (birch profilin). Clinical reactivity to peach was established by double-blind, placebo-controlled food challenges. The clinical characteristics and the in vivo and in vitro tests were compared between allergic and nonallergic patients. RESULTS: Peach allergy was confirmed in 76 patients and ruled out in 16; 2 patients dropped out, and the study was not conclusive in 6 individuals (placebo reactors). Pollen allergy was found in 76% of the allergic patients and in 100% of the nonallergic patients. Positive responses to Pru p 3, rBet v 1, and rBet v 2 were observed in 62%, 7%, and 34% of patients allergic to peach, respectively. The sensitization rate to Pru p 3 was significantly higher among subjects allergic than nonallergic to peach (62% vs 31%, P =.02). IgE responses to rBet v 2 were more frequent among subjects allergic to pollen, but no difference was observed in the presence or absence of peach allergy. CONCLUSIONS: Pru p 3 is the major allergen of peach in our population, and the IgE response to this allergen is related to the clinical expression of peach allergy. Sensitization to profilin is observed in those patients with an associated pollen allergy but does not appear to be related to the clinical reactivity to peach. PMID- 14564366 TI - Acute thromboembolic events associated with intravenous immunoglobulin infusion in antibody-deficient patients. PMID- 14564367 TI - Histamine inhalation challenge in recurrent uvula angioedema. PMID- 14564368 TI - Eosinophilic and neutrophilic sputum response to bronchial challenge with cockroach. PMID- 14564369 TI - Levels of airborne Gly m 1 in regions of soybean cultivation. PMID- 14564370 TI - Food allergy to green tea. PMID- 14564371 TI - Breast-feeding, passive smoking, and asthma and wheeze in children. PMID- 14564373 TI - Prometheus in Gloucestershire: Edward Jenner, 1749-1823. PMID- 14564376 TI - Marked reduction of brainstem blood flow in artificially ventilated newborn piglets during normoxia and normocapnic hypoxia. AB - OBJECTIVE: To estimate the effect of artificial ventilation on regional cerebral blood flow, cardiovascular regulation, and cerebral oxidative metabolism in newborns. DESIGN AND SUBJECTS: Comparison of three randomized treatment groups of newborn piglets: Group 1 (artificially ventilated sham-operated group; n =7); group 2 (artificially ventilated group with normoxia and moderate normocapnic hypoxia; n =7); group 3 (spontaneously breathing group with normoxia and moderate normocapnic hypoxia; n =6). MEASUREMENTS AND RESULTS: Animals were anesthetized with 0.5% isoflurane in 70% nitrous oxide and 30% oxygen. Groups 1 and 2 were artificially ventilated. Animals in group 3 breathed spontaneously. Moderate normocapnic hypoxia was induced in groups 2 and 3 for 1 h by lowering the inspiratory oxygen fraction from 0.35 to 0.11. Mode of ventilation induced at most marginal effects on global cerebrovascular response, cardiovascular regulation, and cerebral oxidative metabolism. However, under normoxic conditions, regional cerebral blood flow of the medulla oblongata, pons, mesencephalon, thalamus, and cerebellum were markedly reduced in artificially ventilated piglets ( P <0.05). Moderate normocapnic hypoxia led to a marked increase in regional cerebral blood flow, which was significantly lower in the medulla oblongata, pons, mesencephalon, thalamus, and cerebellum of artificially ventilated piglets ( P <0.05). CONCLUSION: Artificial ventilation clearly induces reduced neuronal activity in the brain stem and cerebellum of newborn piglets. This is suggested by a considerably reduced blood flow in these regions under normoxia and moderate normocapnic hypoxia. However, there is no relevant detrimental effect on cardiovascular regulation and brain oxidative metabolism. PMID- 14564377 TI - Unusual central venous catheter malposition into the left internal mammary vein: a case report. PMID- 14564380 TI - Serum tumour necrosis factor alpha levels in severe malaria: effect of partial exchange transfusion. PMID- 14564379 TI - Head helmet versus face mask for non-invasive continuous positive airway pressure: a physiological study. AB - OBJECTIVE: To assess selected physiological effects of non-invasive continuous positive airway pressure delivered by head helmet (CPAPH), a special interface device designed to completely contain the head of the patient, compared to face mask (CPAPM). DESIGN: Randomized physiological study. SETTING: University research laboratory. PATIENTS: Eight healthy volunteers. INTERVENTION: Continuous positive airway pressure delivered by face mask and CPAPH in random order. Three gas flow rates (20-30-40 l/min and 30-45-60 l/min, respectively, for CPAPM and CPAPH) and four CPAP levels (0-5-10-15 cmH2O) were employed in a randomized sequence. MEASUREMENTS AND RESULTS: In each patient we monitored airway pressure, esophageal pressure, expiratory flow, and inspiratory and expiratory CO2 concentration. End-expiratory lung volume changes from CPAP 0 were measured by inductance plethysmography. The application of increased levels of CPAP resulted in a significant increase in end-expiratory lung volume, similar for CPAPH and CPAPM. Inspiratory changes of airway pressure were comparable for the two CPAP modes. Inspiratory CO2 concentration was higher during CPAPH (significantly decreased at increased gas flow rates), compared to CPAPM. CONCLUSIONS: Continuous positive airway pressure delivered by head helmet is as effective as CPAPM in increasing end-expiratory lung volume and in compensating for airway pressure changes without the need of a reservoir bag. Higher gas flow rates are necessary to maintain a relatively low inspiratory CO2 concentration. PMID- 14564378 TI - Sleep in the intensive care unit. AB - Abnormalities of sleep are extremely common in critically ill patients, but the mechanisms are poorly understood. About half of total sleep time occurs during the daytime, and circadian rhythm is markedly diminished or lost. Judgments based on inspection consistently overestimate sleep time and do not detect sleep disruption. Accordingly, reliable polygraphic recordings are needed to measure sleep quantity and quality in critically ill patients. Critically ill patients exhibit more frequent arousals and awakenings than is normal, and decreases in rapid eye movement and slow wave sleep. The degree of sleep fragmentation is at least equivalent to that seen in patients with obstructive sleep apnea. About 20% of arousals and awakenings are related to noise, 10% are related to patient care activities, and the cause for the remainder is not known; severity of underlying disease is likely an important factor. Mechanical ventilation can cause sleep disruption, but the precise mechanism has not been defined. Sleep disruption can induce sympathetic activation and elevation of blood pressure, which may contribute to patient morbidity. In healthy subjects, sleep deprivation can decrease immune function and promote negative nitrogen balance. Measures to improve the quantity and quality of sleep in critically ill patients include careful attention to mode of mechanical ventilation, decreasing noise, and sedative agents (although the latter are double-edged swords). PMID- 14564381 TI - Day hospital vs. home treatment--a comparison of illness severity and costs. AB - BACKGROUND: Acute home treatment services, providing short-term intensive input as an alternative to in-patient admission, have been recommended by the Department of Health as part of a spectrum of care. The lack of research evidence for such services is in contrast to acute day hospital care which has been better researched, but not widely adopted. This paper compares the patients treated in a randomised controlled trial (RCT) of day hospital vs. in-patient care with patients treated several years later in the home treatment service which developed from the original acute day hospital. METHOD: In the original RCT, patients were randomised at the point of admission to day hospital or in-patient care. The home treatment sample consisted of a consecutive series of admissions. Severity of illness was assessed at admission using the Comprehensive Psychopathological Rating Scale (CPRS). Both samples were followed up for 12 months to monitor service use and costs. RESULTS: Symptom severity among the home treatment sample (n = 71) was greater than the day hospital sample (n = 94) (mean CPRS score 31.6 vs. 25.5, p < 0.0001). This difference remained significant following adjustment for other socio-demographic and illness variables. Secondary care costs for the home treatment sample (including in-patient, home treatment and out-patient costs) were intermediate between the costs for the day hospital and in-patient samples from the RCT, but the differences were not statistically significant. CONCLUSIONS: Extending the remit of an acute day hospital to provide 24-h care and a choice of treatment location is associated with an increase in the severity of illness treated. The impact on costs is unclear and the total cost of the new service may not be significantly less than in-patient care. The results need to be interpreted with caution because of differences in recruitment methods. PMID- 14564382 TI - First admissions for mood disorders in immigrants to the Netherlands. AB - BACKGROUND: The risk of developing schizophrenia is increased for immigrants to the Netherlands from Surinam, the Netherlands Antilles and Morocco, but not for immigrants from Turkey. We examined, in these groups, the risks of a first admission for manic-depressive psychosis. METHODS: The Dutch Psychiatric Registry provided two datasets. The first referred to first admissions for manic depressive psychosis (MDP), manic or circular type, in the period 1990-1996, the second to first admissions for MDP, depressed type. MDP, depressed type, corresponds (broadly) to the DSM-IV category of major depressive disorder and MDP, manic or circular type, to the DSM-IV category of bipolar I disorder. The Dutch Central Bureau for Statistics provided yearly population figures. RESULTS: There were only small increases in the risks of a first admission for MDP, manic or circular type, for immigrants from Surinam (age- and sex-adjusted RR = 1.14; 95% CI: 0.97-1.33) and the Netherlands Antilles (RR = 1.41; 1.10-1.80). This risk was not clearly increased for immigrants from Morocco. The risks for MDP, depressed type, were increased for males from Morocco (age-adjusted RR = 2.17; 1.72-2.72) and Turkey (RR = 1.83; 1.46-2.30), and significantly decreased for females in all of the immigrant groups. CONCLUSION: We found no evidence for a large increase in the incidence of MDP, manic or circular type, in the immigrant groups, whereas an increase in MDP, depressed type, was observed only in selected groups. Female immigrants suffering from MDP, depressed type, may be underserved. PMID- 14564383 TI - A comparison of anxiety and depressive symptomatology in postpartum and non postpartum mothers. AB - BACKGROUND: The aim of the study was to compare the distribution of symptoms of anxiety and depression among postpartum as compared with non-postpartum mothers, and to estimate the impact of the postpartum period on the risk of anxiety and depression when adjusted for other risk factors. METHODS: A questionnaire survey approaching all women 18-40 years of age in two municipalities in Norway during the period 1998-1999 was conducted. A total of 2,730 women were included, of whom 416 were in the postpartum period. Only women with one or more deliveries (n = 1,794) were included in the multivariate analyses. RESULTS: The crude prevalence of anxiety and depression was lower in postpartum as compared to non-postpartum mothers. However, when controlling for other risk factors, the odds ratio for anxiety was 1.2 (95% CI: 0.6-2.3) and for depression 1.8 (95% CI: 1.1-2.9) during the postpartum period. CONCLUSION: The overall risk of anxiety appeared to be the same in both groups, whereas the risk of depression was increased in the postpartum group. PMID- 14564384 TI - Deceased, disabled or depressed--a population-based 6-year follow-up study of elderly people with depression. AB - The prognosis of depression in elderly people is reported as poor, with high mortality and high rate of non-recovery. The aim of this study was to investigate the outcome after 6 years in a broader perspective. In addition to the risk of dying, we also estimated the risk of moving to long-term care, and having a depression in a re-screening event after 6 years. In 1993, a screening event for depressive symptoms using the Hopkins Symptom Checklist (HSCL-25) was carried out on 1215 subjects aged 65 years and older living in a health care district in Karlstad, Sweden. The study population was divided into a high score, a low score and a dropout group. A re-screening of the same population was carried out in 1999. Logistic regression analysis was used for calculating odds ratios for the selected end-points adjusted for age, gender, civil status and occurrence of physical illness. The adjusted odds ratios for dying for the high score group were 2.5 (95% CI 1.5-4.4) and for any of the end-points 6.1 (95% CI 3.5-10.8) compared with the low score group. Nearly three-quarters of the subjects in the high score group were deceased, depressed or had moved to long-term care after 6 years. The prognosis was poor for depressed elderly people despite not having more physical illness. It is believed that the poor outcome is due to an additive effect of depression on the total medical disease burden. PMID- 14564385 TI - Parental explanatory models of ADHD: gender and cultural variations. AB - BACKGROUND: This study describes parents' explanatory models of Attention Deficit Hyperactivity Disorder (ADHD) and examines model variation by child characteristics. METHOD: Children with ADHD (N = 182) were identified from a school district population of elementary school students. A reliable coding system was developed for parental responses obtained in ethnographic interviews in order to convert qualitative into numerical data for quantitative analysis. RESULTS: African-American parents were less likely to connect the school system to ADHD problem identification, expressed fewer worries about ADHD-related school problems, and voiced fewer preferences for school interventions than Caucasian parents, pointing to a potential disconnect with the school system. CONCLUSIONS: More African-American than Caucasian parents were unsure about potential causes of and treatments for ADHD, indicating a need for culturally appropriate parent education approaches. PMID- 14564386 TI - Public attitudes to schizophrenia in rural Turkey. AB - BACKGROUND: The aim of this study was to determine the public's attitudes and their correlates towards patients with schizophrenia in rural areas. METHODS: The study was carried out in a village near Manisa City, Turkey. Two hundred and eight subjects completed the public survey form which consists of ten items screening demographic and health status and 32 items rating attitudes towards schizophrenia. RESULTS: Half of the subjects stated that persons with schizophrenia are aggressive and that they should not be free in the community. More than half of the subjects stated that they would be irritated about having a neighbour with schizophrenia (61.5%), that they would not rent their home to a person with schizophrenia (58.2%), that they do not want to work with a person with schizophrenia (61.1%), and that they would not get married to a person with schizophrenia (85.6%). CONCLUSIONS: The public in rural areas sufficiently recognises schizophrenia but has a tendency to stigmatise schizophrenic patients. Their attitudes are generally negative and rejective. They do not want close contact with schizophrenic patients. Interpretation of schizophrenia as a mental illness leads to more negative attitudes and increases the social distance. PMID- 14564387 TI - Courtesy stigma--a focus group study of relatives of schizophrenia patients. AB - BACKGROUND: Stigmatization of people with mental illness has been investigated in numerous studies. Little research, however, has been done to explore how relatives of people with schizophrenia perceive and experience stigmatization and how they can fight such stigmatization. METHOD: Aiming to explore stigma from the perspective of relatives of people with schizophrenia, focus group interviews were conducted with 122 members of advocacy groups from different parts of Germany. Focus group sessions were tape- and video-recorded and transcribed. Transcripts were coded using an inductive method, generating categories (domains) from the material. RESULTS: The analysis of focus group data shows that, contrary to previous research findings, discrimination and disadvantages encountered by relatives of schizophrenia patients reach far beyond the spheres of direct social interaction and access to social roles. Our study revealed two additional domains in which relatives encounter stigmatization: structural discrimination and public images of mental illness. Furthermore, psychiatry has been identified as one important source of stigma. Relatives also suggested numerous anti-stigma interventions. These can be grouped into five main categories: communication measures, support for the ill and their relatives, changes in mental health care, education and training, and control and supervision. DISCUSSION: Based on our findings,ways of how relatives of schizophrenia patients and mental health professionals can fight against stigma are discussed. PMID- 14564389 TI - Analysis of eggplant ( Solanum melongena)-related germplasm: morphological and AFLP data contribute to phylogenetic interpretations and germplasm utilization. AB - A total of 94 Solanum accessions, including eggplants and related species, were morphologically characterized based on greenhouse observations, and molecularly analysed by the AFLP technique. Morphological parameters were helpful in assessing similarities or differences among accessions, and molecular data were used to support morphological conclusions. A dendrogram was computed based on the Dice genetic distances using the neighbour-joining method. The analysis was efficient in the assignment of a species name for eight out of nine accessions that were not previously classified, and revealed that 14 further accessions were misnamed in the collection originally received. The results indicate that the taxonomy of Solanum sections and subgenera including several species should be reconsidered. The AFLP technique was revealed as an efficient tool in determining genetic relationships among species. In general, morphological observations were consistent with molecular data, indicating that both approaches complemented to define the phylogenetic status of a large genus like Solanum. In terms of eggplant breeding, the molecular analysis of the Melongena complex, and of the other sections of the subgenus Leptostemonum, establishes useful germplasm relationships in the gene pool available for the genetic improvement of the cultivated species. The results we have provided highlight an urgent necessity to include molecular parameters in handling and characterizing the genebank deposited germplasm related to cultivated crops. PMID- 14564390 TI - About the origin of European spelt ( Triticum spelta L.): allelic differentiation of the HMW Glutenin B1-1 and A1-2 subunit genes. AB - To investigate the origin of European spelt ( Triticum spelta L., genome AABBDD) and its relation to bread wheat ( Triticum aestivum L., AABBDD), we analysed an approximately 1-kb sequence, including a part of the promoter and the coding region, of the high-molecular-weight (HMW) glutenin B1-1 and A1-2 subunit genes in 58 accessions of hexa- and tetraploid wheat from different geographical regions. Six Glu-B1-1 and five Glu-A1-2 alleles were identified based on 21 and 19 informative sites, respectively, which suggests a polyphyletic origin of the A and B-genomes of hexaploid wheat. In both genes, a group of alleles clustered in a distinct, so-called beta subclade. High frequencies of alleles from the Glu-B1 1 and Glu-A1-2 beta subclades differentiated European spelt from Asian spelt and bread wheat. This indicates different origins of European and Asian spelt, and that European spelt does not derive from the hulled progenitors of bread wheat. The conjoint differentiation of alleles of the A- and B-genome in European spelt suggests the introgression of a tetraploid wheat into free-threshing hexaploid wheat as the origin of European spelt. PMID- 14564391 TI - Fine mapping of the parthenocarpic fruit ( pat) mutation in tomato. AB - The parthenocarpic fruit ( pat) gene of tomato is a recessive mutation conferring parthenocarpy, which is the capability of a plant to set seedless fruits in the absence of pollination and fertilization. Parthenocarpic mutants offer a useful method to regulate fruit production and a suitable experimental system to study ovary and fruit development. In order to map the Pat locus, two populations segregating from the interspecific cross Lycopersicon esculentum x Lycopersicon pennellii were grown, and progeny plants were classified as parthenocarpic or wild-type by taking into account some characteristic aberrations affecting mutant anthers and ovules. Through bulk segregant analysis, we searched for both random and mapped AFLPs linked to the target gene. In this way, the Pat locus was assigned to the long arm of chromosome 3, as also confirmed by the analysis of a set of L. pennellii substitution and introgression lines. Afterwards, the Pat position was refined by using simple sequence repeats (SSRs) and conserved ortholog set (COS) markers mapping in the target region. The tightest COSs were converted into CAPS or SCAR markers. At present, two co-dominant SCAR markers encompassing a genetic window of 1.2 cM flank the Pat locus. Considering that these markers are orthologous to Arabidopsis genes, a positional cloning exploiting the tomato- Arabidopsis microsynteny seems to be a short-term objective. PMID- 14564393 TI - Sequence variations of simple sequence repeats on chromosome-4 in two subspecies of the Asian cultivated rice. AB - Computational screening of the chromosome-4 sequence of the rice cultivar Nipponbare ( Oryza sativa L. japonica) revealed 1,844 tandem simple sequence repeats (SSRs) or microsatellites with SSR motifs >/=20 bp and repeated unit length of 1-6 base pairs. Thus SSRs occur once in every 18.8 kb, on the average, on the chromosome with one SSR per 23.8 kb and 16 kb on the short and long arms, respectively. No SSR was detected in the core region of the centromere. Poly(AT)(n) repeats represented the most abundant and length polymorphic class of SSRs on the chromosome, but it did not occur in the exons. GC-rich trinucleotide repeats were most abundant in the coding regions, representing 71.69% of the SSRs identified in the exons. Two hundred and twenty four SSRs were associated with the repetitive DNA sequences, most of them were poly(AT)(n) tracts. Sequence variations of SSRs between two cultivars, representing the two subspecies of the Asian cultivated rice indica and japonica, were identified, revealing that divergence and convergence of the two subspecies could be traced by the analysis of SSRs. These results provide a great opportunity for SSR-based marker development and comparative genome analysis of the two subspecies of the Asian cultivated rice. PMID- 14564392 TI - Factors affecting polymorphism at microsatellite loci in bread wheat [ Triticum aestivum (L.) Thell]: effects of mutation processes and physical distance from the centromere. AB - The effects of factors known to influence the level of polymorphism at microsatellite loci were studied using 99 markers and seven lines of bread wheat. Mutational factors as well as indirect selective events shape diversity at these loci. Theory predicts that the selection of favorable alleles should reduce polymorphism at neutral neighboring loci in genomic areas with low recombination rates. In wheat, local recombination rate is positively correlated with physical distance from the centromere. Seventy four loci among the 99 used could be physically located on the chromosome. We studied how the following affected the diversity among a set of inbred lines: the length of the alleles, the motif (CA versus CT), the structure of the loci (perfect versus imperfect) and the chromosomal position of the loci. For each locus, we determined whether the polymorphism observed at a locus was compatible with the Stepwise Mutation Model (SMM) or the Two-Phase Model (TPM). Both the mutation rate and the compatibility with the SMM or the TPM were shown to be variable between loci. Wheat microsatellite loci were found to be more variable when segregating alleles were perfect and had long motifs (composed of many repetitions). Diversity observed at 19 loci was not compatible with the SMM. Loci located in distal regions, with presumably high recombination rates, had longer allele sizes and were more polymorphic than loci located in proximal regions. We conclude that both mutation factors and indirect selective events vary according to the local recombination rate and therefore jointly influence the level of polymorphism at microsatellite loci in wheat. PMID- 14564394 TI - Tightly linked di- and tri-nucleotide microsatellites do not evolve in complete independence: evidence from linked (TA)n and (TAA)n microsatellites of chickpea (Cicer arietinum L.). AB - In order to understand the dynamics of microsatellite evolution, we have studied allelic variation at a closely linked (TA)(n) and (TAA)(n) microsatellite loci in 114 land races of chickpea ( Cicer arietinum L.), sampled worldwide. These two loci are separated by 27 bp. The two loci showed a very high degree of polymorphism and hence the combined length with the genetic diversity of 0.93, 0.90 and 0.98 for (TAA)(n), (TA)(n) and the combined length, respectively. Using the variation data at the linked loci, a standardized index of linkage disequilibrium was also computed ( I(S)(A)=0.092), which tests the null hypothesis of no linkage and was significant, indicating the presence of linkage disequilibrium. Furthermore, the dynamics of allelic variation showed that there is a threshold combined length, below which both (TAA)(n) and (TA)(n) loci evolve independently, and above which, if one locus increase in size, the other closely linked locus has a tendency to decrease its size and vice versa, without change in the overall ratio of (TAA)(n) and (TA)(n) allele sizes at the region. This result indicates that there are processes in the cell, which 'read' the combined size of the two loci both for proportion and length and determine the direction of tightly linked di- and tri-nucleotide repeat evolution. PMID- 14564396 TI - Construction of a HindIII Bacterial Artificial Chromosome library and its use in identification of clones associated with disease resistance in chickpea. AB - A chickpea ( Cicer arietinum L.) Bacterial Artificial Chromosome (BAC) library from germplasm line, FLIP 84-92C, was constructed to facilitate positional cloning of disease resistance genes and physical mapping of the genome. The BAC library has 23,780 colonies and was calculated to comprise approximately 3.8 haploid-genome equivalents. Studies on 120 randomly chosen clones revealed an average insert size of 100 kb and no empty clones. Colony hybridization using the RUBP carboxylase large subunit as a probe resulted in a very low percentage of chloroplast DNA contamination. Two clones with a combined insert size of 200 kb were isolated after the library was screened with a Sequence Tagged Microsatellite Site (STMS) marker, Ta96, which is tightly linked to a gene ( Foc3) for resistance to fusarium wilt caused by Fusarium oxysporum Schlechtend.: Fr. f. sp. ciceris (Padwick) race 3 at a genetic distance of 1 cM. Also, these two clones were analyzed with several resistance gene analog (RGA) markers. End sequencing of these clones did not identify repetitive sequences. The development of the BAC library will facilitate isolation of Foc3 and allow us to perform physical mapping of this genomic region where additional R genes against other races of the wilt causing pathogen are positioned. PMID- 14564395 TI - Comparative mapping between quercus and castanea using simple-sequence repeats (SSRs). AB - Simple sequence repeat (SSR) markers from Quercus and Castanea were used for comparative mapping between Quercus robur (L.) and Castanea sativa (Mill.). We tested the transferability of SSRs developed in Quercus to Castanea and vice versa. In total, 47% (25) of the Quercus SSRs and 63% (19) of the Castanea SSRs showed a strong amplification product in the non-source species. From these 44 putative comparative anchor tags, 19 (15 from Quercus and 4 from Castanea) were integrated in two previously established genetic linkage maps for the two genera. SSR loci were sequenced to confirm the orthology of the markers. The combined information from both genetic mapping and sequence analysis were used to determine the homeology between seven linkage groups, aligned on the basis of pairs or triplets of common markers, while two additional groups were matched using a single microsatellite marker. Orthologous loci identified between Q. robur and C. sativa will be useful as anchor loci for comparative mapping studies within the Fagaceae family. PMID- 14564397 TI - The genetic basis of stay-green in rice analyzed in a population of doubled haploid lines derived from an indica by japonica cross. AB - Delayed leaf-senescence, or stay-green, has been regarded as a desired characteristic for the production of a number of crops including rice. In this study, we analyzed the genetic basis of stay-green using a population of 190 doubled haploid lines from the cross between an indica parent Zhenshan 97 and a stay-green japonica parent Wuyujing 2. The population was tested in replicated field trials in 2 consecutive years, and six traits were defined to evaluate the stay-green characteristics. A genetic linkage map with 179 SSR (simple sequence repeat) marker loci was constructed. The software QTLMapper, based on a mixed linear model approach, was applied to detect QTLs, epistatic effects and their environmental interactions for these traits. A total of 46 main-effect QTLs was detected for the six traits that can be localized to 25 chromosomal regions. The individual effects of all the QTLs were small. Fifty digenic interactions were resolved that involved 66 loci distributed on all 12 chromosomes. Environmental interactions were detected for 18 of the main-effect QTLs and 14 of the epistatic interactions. Collectively, the epistatic effects and QTL by year interactions accounted for large proportions of the phenotypic variations. The results also showed that most of the stay-green traits were negatively correlated with yield and its component traits. The implications of the results in crop improvement were discussed. PMID- 14564398 TI - A genetic linkage map of the diplosporous chromosomal region in Taraxacum officinale (common dandelion; Asteraceae). AB - In this study, we mapped the diplosporous chromosomal region in Taraxacum officinale, by using amplified fragment length polymorphism technology (AFLP) in 73 plants from a segregating population. Taraxacum serves as a model system to investigate the genetics, ecology, and evolution of apomixis. The genus includes sexual diploid as well as apomictic polyploid, mostly triploid, plants. Apomictic Taraxacum is diplosporous, parthenogenetic, and has autonomous endosperm formation. Previous studies have indicated that these three apomixis elements are controlled by more than one locus in Taraxacum and that diplospory inherits as a dominant, monogenic trait ( Ddd; DIP). A bulked segregant analysis provided 34 AFLP markers that were linked to DIP and were, together with two microsatellite markers, used for mapping the trait. The map length was 18.6 cM and markers were found on both sides of DIP, corresponding to 5.9 and 12.7 cM, respectively. None of the markers completely co-segregated with DIP. Eight markers were selected for PCR-based marker development, of which two were successfully converted. In contrast to all other mapping studies of apomeiosis to date, our results showed no evidence for suppression of recombination around the DIP locus in Taraxacum. No obvious evidence for sequence divergence between the DIP and non- DIP homologous loci was found, and no hemizygosity at the DIP locus was detected. These results may indicate that apomixis is relatively recent in Taraxacum. PMID- 14564399 TI - Genetic mapping in (Populus tomentosa x Populus bolleana) and P. tomentosa Carr. using AFLP markers. AB - The AFLP genetic linkage maps for two poplar cultivars were constructed with the pseudo-test-cross mapping strategy. The hybrids were derived from an interspecific backcross between the female hybrid clone "TB01" (Populus tomentosa x Populus bolleana) and the male clone "LM50" ( P. tomentosa). A total of 782 polymorphic fragments were obtained with a PCR-based strategy using 49 enzyme nested ( EcoRI/ MseI) primer combinations. Six hundred and thirty two of these fragments segregated in a 1:1 ratio ( P<0.01), indicating that these DNA polymorphisms are heterozygous in one parent and null in the other. The linkage analysis was performed using Mapmaker version 3.0 with LOD 5.0 and a maximum recombination fraction (theta) of 0.3. Map distances were estimated using the Kosambi mapping function. In the framework map for "LM50" (P. tomentosa), 218 markers were aligned in 19 major linkage groups. The linked loci spanned approximately 2,683 cM of the poplar genome, with an average distance of 12.3 cM between adjacent markers. For "TB01" (P. tomentosa x P. bolleana), the analysis revealed 144 loci, which were mapped to 19 major linkage groups and covered about 1,956 cM, with an average distance of 13.6 cM between adjacent markers. These maps covered about 87% and 77% of the estimated genome size of parents "LM50" and "TB01", respectively. The maps developed in this study lay an important foundation for future genomics research in poplar, providing a means for localizing genes controlling economically important traits in P. tomentosa. PMID- 14564400 TI - Molecular tagging of erucic acid trait in oilseed mustard (Brassica juncea) by QTL mapping and single nucleotide polymorphisms in FAE1 gene. AB - Molecular mapping and tagging of the erucic acid trait (C22:1) in Brassica juncea was done by a candidate gene approach. Two QTLs underlying the variation of seed erucic acid content were assigned to two linkage groups of a B. juncea map using a doubled haploid (DH) mapping population derived from high x low erucic acid F(1) hybrid. Two consensus primers corresponding to the full-length Fatty Acid Elongase 1 ( FAE1) gene, reported to be involved in the elongation of C18:1 to C22:1, were designed. PCR amplification and subsequent cloning and sequencing identified two FAE1 genes ( FAE1.1 and FAE1.2) in both high and low erucic acid mustard lines. Sequence alignment of corresponding FAE1 genes between high and low erucic acid mustard lines identified four substitution type single nucleotide polymorphisms (SNPs) in FAE1.1 and three in FAE1.2. Using the SNuPE method of SNP genotyping, these two genes were mapped to two independent loci that co segregated with the two QTLs governing the erucic acid trait. Association of wild ( E1E2) and mutant ( e1e2) haplotypes of two FAE1 genes with erucic acid variation in two segregating populations revealed that the e1e1e2e2 genotype identified low erucic acid individuals (<2%) and E1E1E2E2 identified individuals with highest erucic acid content (>40%). The E1e1E2e2 heterozygote was found to be intermediate in phenotype. The applicability of these SNPs in marker-assisted manipulation of the erucic acid trait was verified by genotyping a set of contrasting germplasm of B. juncea belonging to two distinct gene pools (Indian and east European) and other oil-yielding Brassica species. PMID- 14564402 TI - Transmission of ice-nucleating active bacteria from a prey reduces cold hardiness of a predator (Araneae: Theridiidae). AB - The influence of ice-nucleating active (INA) bacteria on cold hardiness of the house spider, Achaearanea tepidariorum, was determined by measuring the supercooling point (SCP) of hatchlings given either INA-bacteria-fed or bacteria free prey (Drosophila melanogaster). Spiders that had eaten INA-bacteria-fed flies showed higher SCPs than those fed on bacteria-free flies. Through feeding, INA bacteria in the prey reduce the cold hardiness of spiders. This fact should be taken into account before using INA agents as a means of pest management. PMID- 14564403 TI - A weevil pollinating the Canary Islands date palm: between parasitism and mutualism. AB - Palm pollination systems are highly diverse, including by wind and by several different groups of insects. Many palm species are associated with more or less specific pollinating weevils that are also floral herbivores. For many such palms, the importance of these "palm flower weevils" as pollinators has not been examined. Here we describe a new ex situ method of demonstrating insect pollination when pollinator exclusion is not possible. We show that Neoderelomus piriformis beetles carry pollen and deposit it on the stigma of Phoenix canariensis. Up until now, pollination systems in Phoenix have been unclear, despite the economic importance of the date palm P. dactylifera. We demonstrate here that small weevils that visit inflorescences and often inconspicuously hide there could be efficient pollinators. PMID- 14564404 TI - Olfactive detection of fig wasps as prey by the ant Crematogaster scutellaris (Formicidae; Myrmicinae). AB - In the species-specific and obligate mutualism between the fig (Ficus carica) and its pollinator (the fig wasps Blastophaga psenes), a third participant, the ant Crematogaster scutellaris, is a predator of the wasps. Here, we ask how ant workers can rapidly localise such prey, whose availability is limited in time and space. Using a Y-tube olfactometer, we tested ant response to odours emitted by different types of figs (receptive female, ripe female or male figs) and by fig wasps (pollinators or non-pollinators). We demonstrate that ants were significantly attracted only to odours emitted by pollinators, either alone or associated with odours of male figs (releasing wasps). Detection of prey odour by ants is an important trait that can explain their observed high rate of predation on pollinators, and could have important implications on the stability of the fig/fig wasp mutualism. PMID- 14564405 TI - Comparative morphology and secretion chemistry of the scoli in caterpillars of Hyalophora cecropia. AB - The morphology of the variously coloured scoli (bristle-bearing structures on the integument, producing an exocrine secretion) on caterpillars of Hyalophora cecropia and the secondary chemistry of the discharged secretions have been investigated for the first time and compared. According to our scanning electron microscopic study, the red/orange, yellow and blue coloured groups of these glands differ morphologically. Gas chromatographic-mass spectrometric analyses showed that the patterns of secondary compounds in the respective glandular secretions are also different. Furthermore, the secretion of the penultimate larval instar is chemically distinct from that of the last instar, as are both secretions from the respective haemolymph. The results favour the idea that the differences in scoli colour, morphology and chemistry could affect various predator species differently. PMID- 14564406 TI - Assortative mating by carotenoid-based plumage colour: a quality indicator in American goldfinches, Carduelis tristis. AB - We investigated the mating patterns in 22 breeding pairs of wild American goldfinches (Carduelis tristis) with respect to their body size, condition, and carotenoid-based, yellow plumage colour. Using reflectance spectrometry, we objectively quantified plumage colours across the bird-visible wavelengths, revealing a unexpected UV peak in the reflectance spectrum from yellow feathers. We summarized our colour measurements using a principal components analysis to create a single variable, carotenoid PC1, that represents the intensity of this carotenoid-based yellow colour, a measure of phenotypic quality in this species. We found no evidence of assortative mating with respect to measures of body size or condition but there was positive assortative mating by carotenoid PC1, such that the yellow plumage colours of males and females were significantly correlated within pairs. We argue that the yellow carotenoid coloration of goldfinches may be important in mutual mate choice and, thus, that sexual selection in this species may act upon female ornamentation, as well as the more obvious plumage signals of males. Because assortative mating results in an increase in genetic variance, we suggest that this might be a mechanism that maintains variance in ornamental traits in spite of the variance-eroding effects of sexual selection. PMID- 14564401 TI - Cellular and molecular mechanisms of embryonic haemangiogenesis and lymphangiogenesis. AB - Vascular diseases are the leading cause of morbidity and mortality in developed nations. Therapeutic haemangiogenesis and lymphangiogenesis may present a chance for curative intervention. Recently we have learned that physiologic and pathologic angiogenesis in the human adult are controlled by molecules that are already functioning during embryogenesis, and the lessons we can learn from studying embryos may be useful for future clinical studies. Here we review cellular and molecular mechanisms of embryonic haemangiogenesis and lymphangiogenesis. The focus lies on the endothelial cell and the surrounding peri-endothelial cells, the growth factors and receptors that control their development, proliferation, maintenance, regression and differentiation. Our rapidly expanding knowledge of the molecules expressed in endothelial cells justifies expanded functional studies in future PMID- 14564407 TI - Biological activities of an extract from Cleome viscosa L. (Capparaceae). AB - Electron micrograph examination of the leaf and stem surfaces of Cleome viscosa L (Family Capparaceae) revealed the presence of secretory glandular trichomes with club-cylinder and cylinder morphologies. In the present study, the leaves and stems of C. viscosa were extracted with hexane and the extract was evaluated for the following biological activities: anti-bacterial, anti-fungal, contact insecticidal and nematicidal. The extract was found to be a potent anti-bacterial agent according to the thin layer chromatography autobiographic assay. Activity directed isolation studies of the anti-bacterially active compounds led to a 14 member ring cembranoid diterpene being identified as one of the effective agents. Minimum inhibitory concentration (MIC) values (microg/spot) of 5.0 microg/spot and 1.0 microg/spot were found for the diterpene on Bacillus subtilis (Gram positive) and Pseudomonas fluorescens (Gram-negative), respectively. The diterpene did not inhibit the growth of the fungus Cladosporium cucumerinum. The extract demonstrated a pyrethroid type of contact insecticidal activity on adult Cylas formicarius elegantulus Summer (Coleoptera: Curculionidae). The extract also had high nematicidal activity with a percentage Abbott's value of 72.69 on the plant parasitic nematode Meloidogyne incognita Chitwood; however, the extract lost its potency upon subfractionation. PMID- 14564408 TI - Early loss and multiple return of the lower temporal arcade in diapsid reptiles. AB - The temporal arches of diapsid reptiles have received attention for several decades. In particular, it has been observed that the lower temporal bar at the ventral margin of the cheek is frequently reduced due to the absence of a contact between jugal and quadratojugal. The loss of the arcade was formerly considered to be of high systematic value, but is now often interpreted as being autapomorphic for the respective taxon, and the presence of both arcades is generally regarded as a plesiomorphic feature. Here I show, based on a cladistic analysis as well as on further anatomical evidence, that the lower temporal arcade was lost only once in diapsid evolution, and that the presence of the arch in "higher" diapsids is secondary, which is indicated by the different ratio between jugal and quadratojugal as well as by ontogeny. This result also sheds new light on the understanding of the cheek configuration of enigmatic taxa such as ichthyosaurs and turtles. PMID- 14564409 TI - The effect of queen pheromones on worker honey bee ovary development. AB - We report results that address a long-standing controversy in honey bee biology, the identity of the queen-produced compounds that inhibit worker honey bee ovary development. As the honey bee is the only organism for which identities have been proposed for any pheromone that regulates reproduction, the resolution of its identity is of broad significance. We examined the effects of synthetic honey bee queen mandibular pheromone (QMP), four newly identified queen retinue pheromone components, and whole-queen extracts on the ovary development of caged worker bees. The newly identified compounds did not inhibit worker ovary development alone, nor did they improve the efficacy of QMP when applied in combination. QMP was as effective as queen extracts at ovary regulation. Caged workers in the QMP and queen extract treatments had better developed ovaries than did workers remaining in queenright colonies. We conclude that QMP is responsible for the ovary-regulating pheromonal capability of queens from European-derived Apis mellifera subspecies. PMID- 14564410 TI - Vision complements echolocation in an aerial-hawking bat. AB - The northern bat Eptesicus nilssonii normally hunts flying insects in the air using frequency-modulated echolocation calls. It is also known to detect and catch visually conspicuous prey (white moths) hovering low among grass stalks. To overcome the problem with acoustic clutter from the grass, which interferes with target echo detection, the bats make use of visual cues in addition to those of echolocation. We therefore investigated the minimum size of prey that the bats could distinguish by using vision, by presenting the bats with different sized dead and spread moths. We found that vision increased the chance of detection only when the moths had a wingspan of at least 5 cm. Smaller targets were detected using echolocation alone. The mean detection range was 3.5 m, suggesting that the bats need a visual acuity of 49' of arc to detect the prey. This is consistent with results of optomotor response tests and counts of retinal ganglion cells in closely related species. Our results suggest that the visual acuity of Eptesicus bats may not be adequate for prey detection under normal conditions, but that the bats can use vision when the prey is unusually large and conspicuous. The northern bats display a flexibility in prey detection techniques not previously recognised among aerial-hawking bats and they are able to use their full visual capacity in the field. PMID- 14564411 TI - Further indications for genetic heterogeneity of euthyroid familial goiter. AB - Iodine deficiency is the most important etiological factor for euthyroid endemic goiter. However, family and twin pair studies also strongly indicate a genetic prediposition. In euthyroid goiters molecular defects in the thyroglobulin (TG), and Na+/I- symporter (NIS) gene have been identified. Numerous mutations in the Pendrin (PDS) gene have been found in families with PDS characterized by deafness and euthyroid goiter. Moreover, family studies indicated two major candidate loci MNG-1 on chromosome 14q31 and Xp22. However, all previous linkage studies investigated only one family. To clarify the general relevance of these previously identified two major candidate loci for the etiology of euthyroid goiter we investigated four families with a total number of 74 family members by linkage analysis with microsatellite markers. Moreover, we analyzed the thyroid candidate genes TG, thyroperoxidase (TPO), NIS, TSH receptor, and PDS. In a further family with 12 members in whom we have previously demonstrated linkage to the MNG-1 locus we investigated the Xp22 locus and the PDS gene in addition to our initial study. Linkage analysis results of our study are not significant enough to definitely exclude or confirm linkage to the investigated candidate genes and loci. Nevertheless, we obtained very weak indications for possible linkage to Xp22 in one family by a maximal multipoint LOD score of 1.15, and cosegregation of haplotypes among affected family members. Moreover, in another family linkage to PDS was indicated by a maximal multipoint LOD score of 1.87 as well as cosegregation of haplotypes. However, sequencing of the PDS gene did not reveal germline mutations. A significant total NPL score of 6.5 for PDS over all families most likely indicated linkage to a genomic region close to PDS. Furthermore, the likelihood of linkage to MNG-1 and Xp22 is reduced, because multipoint LOD scores were below 1 or negative. In all families there was no significant evidence for linkage for the thyroid candidate genes TG, TPO, NIS, or the TSH receptor. In conclusion, a general role of MNG-1 and Xp22 for the etiology of euthyroid goiter is unlikely but cannot clearly excluded. The multipoint parametric and nonparametric LOD scores further suggest genetic heterogeneity in the etiology of familial euthyroid goiter. To identify other susceptibility loci it is necessary to perform genome-wide linkage analysis studies with more families. PMID- 14564413 TI - [Vulnerable plaque -- vulnerable vessel -- vulnerable patient. Plaques prone to rupture in atheroma/fibroatheroma]. PMID- 14564412 TI - A novel mutation, Arg71Thr, in the delta-sarcoglycan gene is associated with dilated cardiomyopathy. AB - Approximately 20-35% of cases of idiopathic dilated cardiomyopathy are familial. DCM-associated mutations have been reported in 13 genes including the desmin, delta-sarcoglycan, and metavinculin genes. This study screened for variants in these genes in Finnish patients with DCM. All coding regions of the desmin and delta-sarcoglycan genes and the metavinculin-specific exon of the vinculin gene were screened in 52 DCM patients from eastern Finland by PCR-SSCP. We detected a novel mutation, Arg71Thr, in the delta-sarcoglycan gene in two members of a small DCM family. One of the mutation carriers fulfills diagnostic criteria for DCM and is also symptomatic. The other mutation carrier has slightly dilated left ventricle and well preserved systolic function. Therefore carriers of the Arg71Thr mutation had a relatively mild phenotype and a late onset of the disease. Disease-associated mutations were not found in the desmin gene or the metavinculin-specific exon of the vinculin gene. We conclude that the desmin and delta-sarcoglycan genes are not predominant disease-causing genes in patients with DCM in eastern Finland. PMID- 14564414 TI - [Magnetic resonance imaging of atherosclerotic plaque]. AB - Recent advances in terms of spatial and temporal resolution have enabled magnetic resonance imaging (MRI) to be used to display atherosclerotic plaque. MRI permits not only to detect atherosclerotic lesions but also enables determination of plaque volume and plaque characterization. It allows to display different plaque components such as lipid core, fibrous cap, calcium, and thrombus. To improve the spatial resolution, different invasive approaches based on intravascular coils have been evaluated. Novel contrast agent developments aim at the detection of inflammatory plaque activity in order to identify lesions with a high vascular risk (vulnerable plaque). PMID- 14564415 TI - Effects induced by mono- and divalent cations on protein regions responsible for thermal adaptation in beta-glycosidase from Sulfolobus solfataricus. AB - The perturbation induced by mono- and divalent cations on the thermophilicity and thermostability of Solfolobus solfataricus beta-glycosidase, a hyperthermophilic tetrameric enzyme, has been investigated by spectroscopic and computational simulation methods to ascertain the Hofmeister effects on two strategic protein regions identified previously. Specifically, (1). an extra segment (83-124), present only in the sequence of hyperthermophilic glycosidases and recognized as an important thermostability determinant for the enzyme structure; and (2). a restricted area of the subunit interface responsible for the quaternary structure maintenance. Mono- and divalent cations inhibit to a different extent the beta glycosidase activity, whose kinetic constants show an apparent competitive inhibition of the catalytic process that reflects the Hofmeister order. The thermostability is also affected by the nature and charge of the cations, reaching maximal effects for the case of Mg(2+). Fourier transform infrared spectroscopy has revealed very small changes in the protein secondary structure in the presence of the investigated cations at 20 degrees C, while large effects on the protein melting temperatures are observed. Computational analysis of the enzyme structure has identified negative patches on the accessible surface of the two identified regions. Following the Hofmeister series, cations weaken the existing electrostatic network that links the extra segment to the remaining protein matrix. In particular, the perturbing action of cations could involve the ionic pair interactions E107-R245 and E109-R185, thus leading to a local destructuring of the extra segment as a possible starting event for thermal destabilization. A detailed investigation of the electrostatic network at the A-C intermolecular interface of Sbetagly after energy minimization suggests that cations could cause a strong attenuation of the ion pair interactions E474-K72 and D473-R402, with consequent partial dissociation of the tetrameric structure. PMID- 14564416 TI - Neuronal and muscular alterations caused by two wheat endosperm proteins, puroindoline-a and alpha1-purothionin, are due to ion pore formation. AB - Using the patch-clamp technique it was found that the toxicity of the two wheat endosperm proteins puroindoline-a and alpha1-purothionin probably results from the dissipation of ion concentration gradients essential for the maintenance of cellular homeostasis. PMID- 14564420 TI - Epidermoid cyst: rare testicular tumor in children. AB - Epidermoid cyst of the testis, extremely rare in children, is a non-teratomatous benign tumor, and testis-sparing surgery should be the treatment of choice. To prevent unnecessary orchiectomy, recognition of this rare tumor in children is essential. PMID- 14564421 TI - Evaluating scrotal imaging: caution advocated. PMID- 14564422 TI - Changes in the size of the lateral ventricles in the normal-term newborn following vaginal delivery. AB - BACKGROUND: This is a study of the size of the lateral ventricles in a population of normal-term newborns following vaginal delivery. OBJECTIVE: To assess the time course for the lateral ventricles to change from closed to open following vaginal delivery. MATERIALS AND METHODS: A total of 143 normal-term newborns had a videotaped cranial ultrasound examination during the period from 1 to 156 h after birth. Many had from one to three additional examinations when returning for well baby checks. Three methods [overlapping intervals (smoothed percentages), life table, and geometric distribution] were used to construct the approximate distributions of completely closed, partially open, and open ventricles. RESULTS: Using smoothed percentages it was estimated that within 12 h of birth, 80% of newborns had closed lateral ventricles, 19% had partially open ventricles, and only 1% were completely open. The estimated median time from birth to partially open ventricles was in the period 36 to 60 h. The life table gave the estimated median time to partially open ventricles as 74 h. Fitting the data to a geometric distribution gave an estimated mean of 63 h. CONCLUSION: The majority of normal newborns have closed lateral ventricles following vaginal delivery and these are partially opened by day 3 after birth. PMID- 14564423 TI - MR imaging of glioblastoma in children: usefulness of diffusion/perfusion weighted MRI and MR spectroscopy. AB - BACKGROUND: Glioblastoma is relatively uncommon in childhood and maybe difficult to differentiate from other brain tumors such as primitive neuroectodermal tumor, ependymoma, or benign astrocytoma. OBJECTIVE: To describe the characteristic MR features in children with glioblastoma and to evaluate the usefulness of diffusion and perfusion MR imaging and MR spectroscopy in pediatric glioblastoma. MATERIALS AND METHODS: MR imaging in 11 children (12 tumors) with biopsy-proven glioblastoma was reviewed retrospectively. In one patient, there was a recurrent glioblastoma. We reviewed CT and MRI imaging for tumor location, density/signal intensity, and enhancement pattern. Routine MR imaging was performed with a 1.5-T scanner. In six patients, diffusion-weighted MR images (DWIs) were obtained with a single-shot spin echo EPI technique with two gradient steps, and apparent diffusion coefficients (ADCs) were calculated. Using the gradient EPI technique, perfusion-weighted MR images (PWIs) were obtained in four patients from the data of dynamic MR images. The maximum relative cerebral blood volume (rCBV) ratio was calculated between the tumor and contralateral white matter in two cases. In three patients, proton MR spectroscopy was performed using a single voxel technique with either STEAM or PRESS sequences. The locations of the tumor were the thalamus and basal ganglia ( n=8), deep white matter ( n=3), and brain stem ( n=1). RESULTS: Intratumoral hemorrhage was seen in four tumors. The tumors showed high-signal intensity or DWIs, having a wide range of ADC values of 0.53-1.30 (mean +/-SD=1.011+/-0.29). The maximum rCBV ratios of glioblastoma were 10.2 and 8.5 in two cases. MR spectroscopy showed decreased N-acetylaspartate (NAA) and increased choline in three cases. The MR findings of glioblastoma in children were: a diffusely infiltrative mass with hemorrhage involving the deep cerebral white matter, thalami, and basal ganglia. CONCLUSION: Diffusion/perfusion MR imaging and MR spectroscopy are very helpful in diagnosing glioblastoma, determining the biopsy site, and evaluating tumor recurrence. PMID- 14564424 TI - Scrotal epidermal inclusion cyst clinically mimicking polyorchidism in a child: ultrasonic characteristics. PMID- 14564425 TI - Association of two respiratory congenital anomalies: tracheal diverticulum and cystic adenomatoid malformation of the lung. AB - Many associations of congenital anomalies of the respiratory system have been reported, but the combination of tracheal diverticulum and cystic adenomatoid malformation (CCAM) is unique. We present a patient with these two anomalies and analyze their embryological correlation. PMID- 14564426 TI - Langerhans' cell histiocytosis of the clivus: case report and literature review. AB - This report describes a 5-year-old girl with Langerhans' cell histiocytosis (LCH) of the clivus. To date only five patients, including our patient, have been described with LCH at this site. Our patient differs from those previously reported by her atypical clinical presentation with torticollis, but without a sixth nerve palsy. In addition, she is the first patient to present with concomitant disease elsewhere at the time of diagnosis, i.e. both femoral necks and left proximal humerus. Our patient thus presents unique features and underlines the importance of including LCH in the differential diagnosis of erosive lesions of the clivus. PMID- 14564427 TI - How to perform the perfect voiding cystourethrogram. AB - The voiding cystourethrogram (VCUG) examination is a difficult investigation to perform and is a stressful experience for patients and their parents, as well as for the radiologists, technicians and paediatric radiology nurses involved in the examination. Despite the VCUG being one of the most commonly performed fluoroscopic procedures in paediatric radiology practice, there is no general consensus as to the best way to perform this investigation. This is particularly concerning when one considers the potentially high gonadal radiation dose children may receive. Because of this, we have undertaken a comprehensive literature review of various aspects of the test in order to determine the best way to perform the VCUG in modern paediatric radiology practice. PMID- 14564428 TI - Imaging of carotid artery disease: from luminology to function? AB - There have been tremendous advances in our ability to image atheromatous disease, particularly in the carotid artery, which is accessible and large enough to image. The repertoire of methodology available is growing, giving anatomical information on luminal narrowing which is approaching the level at which conventional carotid angiography will become very uncommon as CT and contrast enhanced MR angiographic techniques become the norm. More exciting is the tentative ability to perform functional plaque imaging addressing enhancement patterns and macrophage activity using MR or positron-emission tomography techniques. These techniques, once rigorously evaluated, may, in addition to complex mathematical modelling of plaque, eventually allow us to assess true plaque risk. Time will best judge whether we will be able to move from the use of simple luminology to assessment of plaque function. PMID- 14564429 TI - A replicate study design for testing bioequivalence: a case study on two desmopressin nasal spray preparations. AB - OBJECTIVE: The present study was carried out to test bioequivalence between two different desmopressin nasal spray preparations. Due to the high variability of plasma pharmacokinetics of intranasally administered peptides like desmopressin, appropriate study designs are required to assess bioequivalence. Therefore, a single-dose, replicate study design was used to evaluate bioequivalence of two desmopressin nasal sprays. SUBJECTS AND METHODS: Thirty-two healthy male volunteers were enrolled in the study and were randomly assigned to receive the test- and reference drug on two occasions in a 4-period 2-sequence crossover study design. Subjects received a single dose of 20 microg (10 microg per nostril) of desmopressin-acetate per study day separated by wash-out periods of at least 1 week. Desmopressin blood concentrations were measured serially over a 14-h period using a validated radioimmunoassay method. Statistical analysis was initially performed using a complicated mixed-analysis model testing for individual bioequivalence according to recommendations by the Food and Drug Administration. This approach, however, failed to converge with all defined main PK parameters and, thus, a traditional mixed analysis of variance analysis based on population averages was definitely used for testing bioequivalence between study drugs. The procedure of selecting an appropriate statistical analysis for a replicate study design was predefined in the study protocol. RESULTS: The 90% confidence intervals (CI) were calculated for the area under the time concentration curve (AUC), maximum concentration (C(max)) and the time to reach C(max) (t(max)) of test/reference drug ratios for a bioequivalence range from 0.80-1.25. The mean test/reference drug ratios were completely within the 90% CIs with values of 1.041 (CI: 0.892-1.216), 1.021 (CI: 0.913-1.140) and 1.068 (CI: 0.914-1.249) for AUC(0-14 h), C(max) and t(max), respectively. CONCLUSION: The rate and the extent of intranasal desmopressin absorption are identical for both study preparations. Thus, the desmopressin test preparation met all equivalence criteria and thereby was proven bioequivalent with a marketed reference nasal desmopressin spray. PMID- 14564430 TI - Metastable equilibrium solubility behavior of carbonated apatite in the presence of solution strontium. AB - The purpose of this study was to use the concept of metastable equilibrium solubility (MES) to describe the anomalous solubility behavior of carbonated apatite (CAP) in the presence of solution strontium. A CAP sample (4.8 wt% CO(3), synthesized at 70 degrees C) was prepared by precipitation. Baseline MES distributions were determined in a series of 0.1 M acetate buffers containing only calcium and phosphate (no strontium) over a broad range of solution conditions. In order to assess the influence of strontium, MES profiles were then determined in a similar fashion with 20, 30, 40, 50, 60, 70, and 80% of the solution calcium being replaced on an equal molar basis by solution strontium. From the compositions of the equilibrating buffer solutions, ion activity products (IAPs) of the form Ca(10-n)Sr(n)(PO(4))(6)(OH)(2) (n = 0-10) were calculated in an attempt to determine the correct function governing the dissolution of the CAP preparation. The results demonstrate the following important findings: (a) at high solution strontium/calcium ratios (i.e., when 60% or more of the solution calcium was replaced by strontium), the MES profiles in all the experiments were found to be essentially superimposable when the solution IAPs were calculated using the stoichiometry of Ca(6)Sr(4)(PO(4))(6)(OH)(2), and (b), at low solution strontium/calcium ratios (i.e., when 40% or less of the solution calcium was replaced by strontium), the stoichiometry yielding MES data superpositioning was found to be that of hydroxyapatite. When other stoichiometries were assumed, good superpositioning of the data was not possible. PMID- 14564431 TI - Type II benign osteopetrosis (Albers-Schonberg disease) caused by a novel mutation in CLCN7 presenting with unusual clinical manifestations. AB - A 16-year-old male patient with type II autosomal dominant benign osteopetrosis (ADO) was genotyped and found to harbor a novel mutation in exon 25 of the gene encoding for the osteoclast-specific chloride channel, CLCN7, inherited from the father, who was asymptomatic. The patient had normal biochemical findings and acid-base balance, except for increased serum levels of creatine kinase, lactic dehydrogenase, and the bone formation markers bone alkaline phosphatase isoenzyme, osteocalcin and N-terminal type I collagen telopeptide/creatinine ratio. Unusual generalized osteosclerosis was observed together with a canonical increase in vertebral and pelvis bone mass. An affected first grade cousin presented with normal biochemical findings and a milder osteosclerotic pattern of the pelvis. At the cellular level, cultured osteoclasts from the patient showed increased motility, with lamellipodia, membrane ruffling and motile pattern of podosome distribution, all of which could have contributed to functional impairment of bone resorption. The present report documents a novel mutation of the CLCN7 gene causing osteopetrosis in a radiologically uncertain form of the diseases, with apparent incomplete penetrance. PMID- 14564432 TI - Subchondral bone reaction associated with chondral defect and attempted cartilage repair in goats. AB - Repair of cartilage damage with autologous chondrocyte transplantation (ACT) has become popular in clinical use during the past few years. Although clinical results have mostly been successful, several unanswered questions remain regarding the biological mechanism of the repair process. The aim of this study was to develop a goat model for ACT. The repair was not successful due to the graft delamination, but we characterize the subchondral changes seen after the procedure. A chondral lesion was created in 14 goat knees, operated on 1 month later with ACT, and covered with periosteum or a bioabsorbable poly-L/D-lactide scaffold. After 3 months, only two of the five lesions repaired with ACT showed partly hyaline-like repair tissue, and all lesions (n = 4) with the scaffold failed. Even though the lesions did not extend through the calcified cartilage, the bone volume and collagen organization of bone structure were decreased when assessed by quantitative polarized light microscopy. There was a significant loss of bone matrix and distortion of the trabecular structure of subchondral bone, which extended several millimeters into the bone. The subchondral bone demonstrated strong hyaluronan staining in the bone marrow and cartilaginous areas with signs of endochondral ossification, suggesting structural remodeling of the bone. The goat model used here proved not to be an optimal model for ACT. The changes in subchondral bone may alter the biomechanical properties of the subchondral plate and thus the long-term survival of the repair tissue after ACT. PMID- 14564433 TI - Effects of weekly administrations of alendronate+clodronate on young mouse tibia: localized action at the proximal growth plate. AB - Alendronate (A), a typical aminobisphosphonate (aminoBP), has a strong bone resorption-inhibitory activity (BRIA). However, like other aminoBPs it has inflammatory side effects. In contrast, the BRIA of clodronate (C), a non aminoBP, is much weaker, and in animal experiments it suppresses aminoBP-induced inflammatory reactions. In the present study, we examined the effects of weekly administrations of A (1.6 micro mol/kg) + C (160 micro mol/kg) on the tibias in young mice and compared them to those induced by A or C alone. Radiophotography showed that A increased bone density at a selective site in the tibia. Indeed, one week after the final injection of A (given alone), clear sclerotic lines (tentatively called BP-lines) were visible at sites corresponding to the location of the growth plate at the time of the each injection. C also produced BP-lines, although they were weaker than those produced by A. Combined administration of A and C produced similar BP-lines as seen in mice given A alone. These results together with other physicochemical effects of A on the tibia suggest that (1) each injection of A and C inhibits bone resorption selectively and transiently at the tibial growth plate in young mice, although minor effects on other sites cannot be excluded, and (2) the combination of A and C keeps still a strong BRIA. Our findings may suggest a strategy for the prevention or reduction of some inflammatory side effects of A or other aminoBPs. PMID- 14564434 TI - Compatibility of staining protocols for bone tissue with Raman imaging. AB - We report the use of Raman microscopy to image mouse calvaria stained with hematoxylin, eosin and toluidine blue. Raman imaging of stained specimens allows for direct correlation of histological and spectral information. A line-focus 785 nm laser imaging system with specialized near-infrared (NIR) microscope objectives and CCD detector were used to collect approximately 100 x 450 micro m Raman images. Principal components analysis, a multivariate analysis technique, was used to determine whether the histological stains cause spectral interference (band shifts or intensity changes) or result in thermal damage to the examined tissue. Image analysis revealed factors for tissue components and the embedding medium, glycol methacrylate, only. Thus, Raman imaging proved to be compatible with histological stains such as hematoxylin, eosin and toluidine blue. PMID- 14564437 TI - An overview of qualimetric strategies for optimisation and calibration in pharmaceutical analysis using flow injection techniques. AB - Flow Injection analysis represents an attractive tool because of its great advantages, such as versatility, speed, high sampling rate and wide applicability in the field of pharmaceutical analysis. However, due to the inherent characteristics of the technique, the choice of the best set of operational and chemical conditions is complicated and the conventional univariate optimisation method present some limitations, mainly due to fact that the interdependence of variables is not considered. In relation to the calibration process, because of the transient character of the signals obtained using FIA manifolds coupled with different detection techniques, different strategies can be used in calibration to solve some problems related to the nature of the signal thereby improving the performance characteristics of the method. This paper offers an overview of different methodologies used in optimisation based on the use of statistically designed experiments and some strategies developed for calibration applied to the analysis of pharmaceuticals. PMID- 14564438 TI - The chromatographic behavior of group (IIB) metal ions on polyurethane foam functionalized with 8-hydroxyquinoline. AB - Polyurethane foam functionalized with 8-hydroxyquinoline has been prepared by coupling the foam matrix with 8-hydroxyquinoline (oxine) through an azo spacer. The oxine-bonded foam (Ox PUF) was characterized by use of different tools (UV Vis spectra, IR spectra, density, and stability). Ox PUF was found to be very suitable for separation and preconcentration of trace metals, e.g. Zn(II), Cd(II), and Hg(II) ions, from wastewater in the pH ranges 2-12, 9-12, and 3-6, respectively. Various conditions influencing the sorption of these metal ions on to Ox PUF were optimized. Extraction of the metal ions was accomplished in 15 to 20 min. Study of the variation of the sorption of the tested metal ions with temperature yielded average values for DeltaH, DeltaS, and DeltaG of 41.99, 158.23, and -5.1 kJ mol(-1), respectively. The capacities of the foam material were 0.27, 0.16, and 0.09 mmol g(-1) for Zn(II), Cd(II), and Hg(II), respectively. Preconcentration factors >50 were achieved (RSD approximately 6.18). The quantitative results were obtained from experiments performed using certified reference materials. PMID- 14564435 TI - Shoulder muscle activity in Parkinson's disease during multijoint arm movements across a range of speeds. AB - Bradykinesia is one of the primary symptoms of Parkinson disease and leads to significant functional limitations for patients. Single joint movement studies, that have investigated the mechanism of bradykinesia, suggest that several features of muscle activity are disrupted, including modulation of burst amplitude and duration, and the number of bursts. It has been proposed that it is the blending of these different burst deficits that collectively defines bradykinesia. This study adds two new approaches to the study of bradykinesia. First, we examined the features of shoulder muscle activities during multijoint arm movement in bradykinetic and control subjects, such that previously reported single joint hypotheses could be tested for generalized arm movement. Second, we directly manipulated speed while keeping distance constant for a large range of speeds. In this manner, we could compare individual trials of muscle activity between controls and subjects with Parkinson's disease (PD) for movements matched for both speed and movement duration. Our results showed that while a multiple burst pattern of shoulder muscles was a common strategy for all subjects (young, elderly controls and PD), subjects with PD showed several burst abnormalities, including deficits in initial agonist burst amplitude and duration at both fast and slow speeds. Subjects with PD also (1) failed to produce a one-burst pattern at fast speeds and, instead, produced a predominance of multiple burst patterns and (2) showed a relationship between the number of burst deficits and the severity of disease. These results extend the findings of single joint studies to multi-joint and similarly indicate that a combination of burst modulation abnormalities correlate with bradykinesia and disease severity. PMID- 14564439 TI - In-vitro assays for determination of oestrogenic activity. AB - At present, in-vitro bioassays are predominantly being seen as tools to identify, through screening programs, whether or not individual chemical compounds have an effect on the endocrine system. However, as the techniques mature, they are likely to find use in the future in monitoring of discharges to the environment for any biological effect and will compliment the range of chemical and biological techniques also available for monitoring environmental quality. Such an approach has already been utilised by a number of workers to fractionate mixtures (e.g. final effluents from STW), to isolate the oestrogenically active components and subsequently identify the compounds which are active. This paper reviews the present state of in-vitro techniques for determination of oestrogenic activity and discusses present approaches to their use in environmental monitoring in conjunction with chemical analyses in toxicity identification and evaluation. PMID- 14564440 TI - Immunochemical determination of xenobiotics with endocrine disrupting effects. AB - This paper is a review with more than 100 references discussing the immunochemical methods reported in the literature for the most important man-made chemicals with suspected endocrine disrupting activity. Details regarding immunizing hapten design, antibody production, and the features (limit of detection, dynamic range, specificity) of the most important immunochemical methods developed (ELISA, FIIA, immunosorbents, immunosensors, etc.) are presented for important environmental pollutants such as bisphenol A, phthalates, alkylphenol polyethoxylates, alkylphenols, polychlorinated biphenyl compounds, and dioxins. Availability of commercial reagents and methods is reported. PMID- 14564441 TI - Direct solid analysis of powdered tungsten carbide hardmetal precursors by laser induced argon spark ablation with inductively coupled plasma atomic emission spectrometry. AB - The potential of the laser-induced argon spark atomizer (LINA-Spark atomizer) coupled with ICP-AES as a convenient device for direct analysis of WC/Co powdered precursors of sintered hardmetals was studied. The samples were presented for the ablation as pressed pellets prepared by mixing with powdered silver binder containing GeO2 as internal standard. The pellets were ablated with the aid of a Q-switched Nd:YAG laser (1064 nm) focused 16 mm behind the target surface with a resulting estimated power density of 5 GW cm(-2). Laser ablation ICP-AES signals were studied as a function of ablation time, and the duration of time prior to measurement (pre-ablation time) which was necessary to obtain reliable results was about 40 s. Linear calibration plots were obtained up to 10% (m/m) Ti, 9% Ta and 3.5% Nb both without internal standardization and by using germanium as an added internal standard or tungsten as a contained internal standard. The relative uncertainty at the centroid of the calibration line was in the range from +/- 6% to +/- 11% for Nb, Ta and Ti both with and without internal standardisation by Ge. A higher spread of points about the regression was observed for cobalt for which the relative uncertainty at the centroid was in the range from +/- 9% to +/- 14%. Repeatability of results was improved by the use of both Ge and W internal standards. The lowest determinable quantities calculated for calibration plots were 0.060% Co, 0.010% Nb, 0.16% Ta and 0.030% Ti with internal standardization by Ge. The LA-ICP-AES analyses of real samples led to good agreement with the results obtained by solution-based ICP determination with a relative bias not exceeding 10%. The elimination of the dissolution procedure of powdered tungsten (Nb, Ta, Ti) carbide is the principal advantage of the developed LA-ICP-AES method. PMID- 14564442 TI - A rapid two-step chromatographic method for the quantitative determination of vitellogenin in fish plasma. AB - By combining anion-exchange membrane purification with high-performance size exclusion chromatography (HPSEC) analysis, a two-step chromatographic method was developed for the determination of vitellogenin (Vtg) in fish plasma. Most plasma protein interferences can be removed during anion-exchange membrane purification process. Vtg is eluted from the size-exclusion chromatography column with a retention time of about 9 min and is characterized based on the native molecular weight, with a limit of quantification of 20 microg Vtg mL(-1)plasma. The spiked recovery and interassay variability were better than 80% and 4.8%. This method was successfully applied to analyze the plasma Vtg levels of loach (Misgurnus angaillicaudatus) and sea catfish (Enchelyopus elongatus). In addition to all the female fish, Vtg is detected in 75% of male loaches and 100% of male sea catfish. The result indicates that some chemicals or unknown factors with estrogenic activity have induced male fish to produce Vtg. PMID- 14564444 TI - Electrochemical studies and square-wave voltammetric determination of fenofibrate in pharmaceutical formulations. AB - The electrochemical reduction of fenofibrate at a hanging mercury drop electrode (HMDE) was investigated by cyclic voltammetry, square-wave voltammetry, and chronoamperometry. Different buffer solutions were used over a wide pH range (3.0 10.0). The best definition of the analytical signals was found in borate buffer (pH 9.0)-tetrabutylammonium iodide mixture containing 12.5% (v/v) methanol at 1.2 V (versus Ag/AgCl). According to cyclic voltammetric studies, the reduction was irreversible and diffusion controlled. The diffusion coefficient was 2.38x10( 6) cm2 s(-1) as determined by chronoamperometry. Under optimized conditions of square-wave voltammetry, a linear relationship was obtained between 0.146-4.96 microg mL(-1) of fenofibrate with a limit of detection of 0.025 microg mL(-1). Validation parameters such as sensitivity, accuracy, precision, and recovery were evaluated. The proposed method was applied to the determination of fenofibrate in pharmaceutical formulations. The results were compared with those obtained by a published high-performance liquid chromatography method. No difference was found statistically. PMID- 14564443 TI - Xenoestrogens: mechanisms of action and detection methods. AB - Estrogenic compounds exert pleiotropic effects in wildlife and humans, and endogenous estrogens, like 17 beta-estradiol, regulate growth and development of their target tissues. Environmental, industrial, or naturally occurring chemicals that possess estrogenic and/or antiestrogenic activities are termed xenoestrogens and may interfere with endocrine systems. These xenoestrogens are therefore defined as endocrine-active or endocrine-disrupting compounds. The estrogen receptor (ER) is the major regulatory unit within the estrogen-signaling pathway and the molecular mechanisms of estrogen and ER actions are described briefly. Based on the mechanism of ER action, in vitro test systems are described that can be employed for screening but also for the elucidation of mechanisms of action of (anti)estrogenic compounds. How screening assays and mechanistic studies can aid in human risk assessment for potential endocrine-active compounds is discussed also. PMID- 14564445 TI - Liquid chromatography with triple-quadrupole or quadrupole-time of flight mass spectrometry for screening and confirmation of residues of pharmaceuticals in water. AB - LC-MS-MS has been performed with triple-quadrupole (QqQ) and quadrupole-time of flight (Q-ToF) instruments and has been used for screening and confirmation of pharmaceuticals in surface, drinking, and ground water. Screening was based on monitoring of one specific MS-MS ion of the target compounds. Confirmation of the identity of the pharmaceuticals was based either on the monitoring of two specific MS-MS ions and calculation of the ratio of their intensities, or on the exact masses of MS-MS product ions obtained for a molecular ion by use of LC-Q ToF MS. The set of pharmaceuticals included four analgesics (acetylsalicylic acid, diclofenac, ibuprofen, and paracetamol), three antibiotics (sulfamethoxazole, erythromycin, and chloramphenicol), five blood-lipid regulators and beta-blockers (fenofibrate, bezafibrate, clofibric acid, bisoprolol, and metoprolol), and the anti-epileptic drug carbamazepine. Limits of quantification ranged from 5 to 25 ng L(-1). Fifty-six samples were analysed and residues of the pharmaceuticals were detected in almost all surface and groundwater and in some drinking water samples. The identity of the compounds could be confirmed by use of both QqQ- and Q-ToF-based LC-MS-MS. However, the latter technique has the distinct advantage that a large number of pharmaceuticals can be screened and confirmed at low concentrations (1-100 ng L( 1)) in one run. PMID- 14564446 TI - Bioimaging in drug discovery sciences. PMID- 14564447 TI - A new approach for assessing early osteoarthritis in the rat. AB - Several animal models have been developed to investigate osteoarthritis and potential disease-modifying therapeutics. However, early disease data from these models are limited by the resolution of current imaging modalities. In this in vitro study, an optical coherence tomography (OCT) system with an axial resolution of 15 micro m was used to track sequential changes in osteoarthritic rat knees. Osteoarthritis was induced via transection of the medial collateral ligament and an artificial full thickness meniscal tear. Imaging occurred at one, two, and three weeks after surgery. OCT successfully detected early signs of osteoarthritic change, including alteration of the cartilage surface and disruption of the bone-cartilage interface. This study demonstrates that OCT, along with the induction of mechanical injury, provides an excellent model for monitoring the sequential changes of osteoarthritis. PMID- 14564448 TI - The effects of hydrocortisone on rat heart muscarinic and adrenergic alpha 1, beta 1 and beta 2 receptors, propranolol-resistant binding sites and on some subsequent steps in intracellular signalling. AB - Glucocorticoids affect the expression and density of neurotransmitter receptors in many tissues but data concerning the heart are contradictory and incomplete. We injected rats with hydrocortisone for 1-12 days and measured the densities of cardiac muscarinic receptors, alpha(1)-, beta(1)- and beta(2)-adrenoceptors and propranolol-resistant binding sites (formerly assumed to be the putative beta(4) adrenoceptor). Some aspects of intracellular signalling were also evaluated: we measured adenylyl cyclase activity (basal, isoprenaline- and forskolin-stimulated and carbachol-inhibited), the coupling between muscarinic receptors and G proteins and basal and isoprenaline-stimulated heart rate. The density of cardiac muscarinic receptors increased (in both the atria and the ventricles). The density of beta(1)-adrenoceptors increased in the atria and was little changed in the ventricles. The density of beta(2)-adrenoceptors increased in both the atria and the ventricles. The number of alpha(1)-adrenoceptors decreased initially, followed by a transient increase in the atria and did not change in the ventricles. The density of propranolol-resistant binding sites first increased and then diminished in the atria and did not change in the ventricles. Although there were noticeable changes in receptor densities, the stimulatory and inhibitory effects on adenylyl cyclase, basal and isoprenaline-stimulated heart rate and the coupling between muscarinic receptors and G proteins were not significantly altered. This may indicate that changes in receptor densities might be one of the mechanisms maintaining stable functional output. PMID- 14564450 TI - Effects of ouabain on human bronchial muscle in vitro. AB - The effects of ouabain, an inhibitor of the plasmalemmal Na(+)/K(+)-ATPase activity, were examined in human isolated bronchus. Ouabain produced concentration-dependent contraction with -logEC(50)=7.16+/-0.11 and maximal effect of 67+/-4% of the response to acetylcholine (1 mM). Ouabain (10 microM) induced contraction was epithelium-independent and was not depressed by inhibitors of cyclooxygenase and lipoxygenase, antagonists of muscarinic, histamine H(1)-receptors and alpha-adrenoceptors, or neuronal Na(+) channel blockade. The inhibition of ouabain contraction in tissues bathed in K(+)-free medium, and the inhibition by ouabain of the K(+)-induced relaxation confirm that the contractile action of ouabain is mediated by inhibition of Na(+)/K(+)-ATPase. Furthermore, depolarization (16.4+/-0.9 mV) was observed in human isolated bronchus by intracellular microelectrode recording. Ouabain (10 microM)-induced contractions were abolished by a Ca(2+)-free solution but not by blockers of L type Ca(2+) channels. In human cultured bronchial smooth muscle cells, ouabain (10 microM) produced a sustained increase in [Ca(2+)](i) (116+/-26 nM) abolished in Ca(2+)-free medium. Incubation with a Na(+)-free medium or amiloride (0.1 mM) markedly inhibited the spasmogenic effect of ouabain thus suggesting the role of Na(+)/Ca(2+) exchange in ouabain contraction while selective inhibitors of Na(+)/H(+)-antiport, Na(+)/K(+)/Cl(-)-antiport, or protein kinase C had no effect. Ouabain (10 microM) failed to increase inositol phosphate accumulation in human bronchus. Ouabain (10 microM) did not alter bronchial responsiveness to acetylcholine or histamine but inhibited the relaxant effects of isoprenaline, forskolin, levcromakalim, or sodium nitroprusside. These results indicate that ouabain acts directly to produce contraction of human airway smooth muscle that depends on extracellular Ca(2+) entry unrelated to L-type channels and involving the Na(+)/Ca(2+)-antiporter. PMID- 14564449 TI - Co-administration of dextromethorphan with morphine attenuates morphine rewarding effect and related dopamine releases at the nucleus accumbens. AB - Morphine is one of the most effective analgesics in clinic to treat postoperative pain or cancer pain. A major drawback of its continuous use is the development of tolerance and dependence. In our previous study we found that a widely used antitussive agent in clinics, dextromethorphan [(DM); also known as a non competitive N-methyl- d-aspartate (NMDA) antagonist], could prevent the development of morphine tolerance. In the present study, we further investigated its effect on morphine addiction. Conditioned place preference (CPP) test and behavioral sensitization of locomotor activity were used to investigate the drug seeking related behaviors, which were in correlation with psychological dependence. Our results showed that co-administered DM was able to abolish completely the CPP effect induced by morphine, but had no effect on morphine induced behavioral sensitization. By employing the microdialysis technique in free-moving animals, we also determined the extracellular level of dopamine and serotonin metabolites in the shell region of the nucleus accumbens (NAc) in its response to morphine with/without DM. A significant increase in dopamine metabolites following morphine administration was demonstrated in the NAc. This increase by morphine could be attenuated by co-administered DM, whereas DM itself did not show any effect. Based on our results, it is speculated that DM may effectively attenuate morphine-induced psychological dependence. Neurochemical analysis revealed that the effect of DM could be through its action on the dopaminergic mesolimbic pathway, which could be activated by morphine and attributed to the cause of rewarding. PMID- 14564452 TI - Dehalorespiration with hexachlorobenzene and pentachlorobenzene by Dehalococcoides sp. strain CBDB1. AB - The chlororespiring anaerobe Dehalococcoides sp. strain CBDB1 used hexachlorobenzene and pentachlorobenzene as electron acceptors in an energy conserving process with hydrogen as electron donor. Previous attempts to grow Dehalococcoides sp. strain CBDB1 with hexachlorobenzene or pentachlorobenzene as electron acceptors failed if these compounds were provided as solutions in hexadecane. However, Dehalococcoides sp. strain CBDB1 was able to grow with hexachlorobenzene or pentachlorobenzene when added in crystalline form directly to cultures. Growth of Dehalococcoides sp. strain CBDB1 by dehalorespiration resulted in a growth yield ( Y) of 2.1+/-0.24 g protein/mol Cl(-) released with hexachlorobenzene as electron acceptor; with pentachlorobenzene, the growth yield was 2.9+/-0.15 g/mol Cl(-). Hexachlorobenzene was reductively dechlorinated to pentachlorobenzene, which was converted to a mixture of 1,2,3,5- and 1,2,4,5 tetrachlorobenzene. Formation of 1,2,3,4-tetrachlorobenzene was not detected. The final end-products of hexachlorobenzene and pentachlorobenzene dechlorination were 1,3,5-trichlorobenzene, 1,3- and 1,4-dichlorobenzene, which were formed in a ratio of about 3:2:5. As reported previously, Dehalococcoides sp. strain CBDB1 converted 1,2,3,5-tetrachlorobenzene exclusively to 1,3,5-trichlorobenzene, and 1,2,4,5-tetrachlorobenzene exclusively to 1,2,4-trichlorobenzene. The organism therefore catalyzes two different pathways to dechlorinate highly chlorinated benzenes. In the route leading to 1,3,5-trichlorobenzene, only doubly flanked chlorine substituents were removed, while in the route leading to 1,3-and 1,4 dichlorobenzene via 1,2,4-trichlorobenzene singly flanked chlorine substituents were also removed. Reductive dehalogenase activity measurements using whole cells pregrown with different chlorobenzene congeners as electron acceptors indicated that different reductive dehalogenases might be induced by the different electron acceptors. To our knowledge, this is the first report describing reductive dechlorination of hexachlorobenzene and pentachlorobenzene via dehalorespiration by a pure bacterial culture. PMID- 14564451 TI - Alpha-naphthoflavone induces vasorelaxation through the induction of extracellular calcium influx and NO formation in endothelium. AB - The effect of alpha-naphthoflavone (alpha-NF) on vascular function was studied in isolated ring segments of the rat thoracic aorta and in primary cultures of human umbilical vein endothelial cells (HUVECs). alpha-NF induced concentration dependent relaxation of the phenylephrine-precontracted aorta endothelium dependently and -independently at lower and higher concentrations, respectively. The cGMP, but not cAMP, content was increased significantly in alpha-NF-treated aorta. Pretreatment with N(omega)-nitro- l-arginine methyl ester (L-NAME) or methylene blue attenuated both alpha-NF induced vasorelaxation and the increase of cGMP content significantly. The increase of cGMP content induced by alpha-NF was also inhibited by chelating extracellular Ca(2+) with EGTA. These results suggest that the endothelium-dependent vasorelaxation induced by alpha-NF is mediated most probably through Ca(2+)-dependent activation of NO synthase and guanylyl cyclase. In HUVECs, alpha-NF induced concentration-dependent formation of NO and Ca(2+) influx. alpha-NF-induced NO formation was abolished by removal of extracellular Ca(2+) and by pretreatment with the Ca(2+) channel blockers SKF 96365 and Ni(2+), but not by the L-type Ca(2+) channel blocker verapamil. The Ca(2+) influx, as measured by (45)Ca(2+) uptake, induced by alpha-NF was also inhibited by SKF 96365 and Ni(2+). Our data imply that alpha-NF, at lower concentrations, induces endothelium-dependent vasorelaxation by promoting extracellular Ca(2+) influx in endothelium and the activation of the NO-cGMP pathway. PMID- 14564456 TI - Heat-induced immunoreactivity of tau protein in neocortical neurons of fire fatalities. AB - Tau protein is the main component of neurofibrillary tangles of Alzheimer's disease (AD). Immunohistochemistry of tau protein is useful in the diagnosis of AD but produces diffuse staining of neocortical neurons in fire fatalities. To learn the cause of this phenomenon, we examined the temporal neocortex of 13 fire fatalities and 9 fatalities unrelated to fire. The diffuse tau immunoreactive neurons were observed in 10 fire fatalities with heat coagulation of the cerebrum. Diffuse staining was not found in the three fire fatalities without heat coagulation of the cerebrum or in fatalities unrelated to fire. The immunoreactivity progressively increased as a function of the degree of cerebral heat coagulation. These results demonstrate that diffuse tau immunoreactivity of neocortical neurons is a post-mortem phenomenon caused by prolonged exposure of the head to intense heat. Forensic pathologists should consider this phenomenon when they diagnose AD in fire fatalities. PMID- 14564455 TI - Assessment of the tibia using ultrasonic guided waves in pubertal girls. AB - The purpose of this study was to compare low frequency ultrasonic guided wave measurements with established ultrasound and bone density measurements in terms of their ability to characterize the tibia in pubertal girls. Subjects were 12-14 year-old girls ( n=106) who were participating in a calcium and vitamin D intervention study. A prototype low frequency pulse transmission device consisting of a uniaxial scanning mechanism and low frequency transducers orientated perpendicularly to the limb was used to measure two ultrasound velocities in the tibia. The first velocity, V1, was that of the first arriving signal, similar to that measured by existing commercial tibial ultrasound devices. The second velocity, V2, was that of a slower wave propagating at 1,500 2,000 m/s, which has been shown elsewhere to be consistent with the lowest order antisymmetric guided mode in the bone. In addition, commercial ultrasound devices (Omnisense, Sunlight Ltd.; QUS-2, Quidel Corp.) were used to measure the speed of sound (SOS) in the tibia and the radius and attenuation (BUA) in the calcaneus. Cortical bone cross-sectional area (CSA), mineral density (BMD) and cortical thickness (cTh) of the tibia were measured using pQCT, site-matched to the ultrasound measurements. Both V1 and V2 correlated significantly with cortical BMD and with cTh and CSA. On the other hand, tibial SOS correlated with BMD, but not with cTh and CSA. These results indicate that the prototype device using guided waves captures aspects of tibial cortical bone geometry in addition to bone density, thereby potentially offering increased diagnostic information compared to existing tibial ultrasound devices. PMID- 14564457 TI - Postmenopausal bleeding: squamous cell carcinoma of cervix with coexisting endometrial tuberculosis. AB - CASE REPORT: A case of squamous cell carcinoma of cervix co-existent with endometrial tuberculosis presenting as postmenopausal bleeding is being reported for its rarity. The atrophic postmenopausal endometrium is thought to be poorly supportive of tubercle bacilli. Following a radical Wertheim's hysterectomy patient had a hectic postoperative period, which responded to antitubercular treatment. Diagnosis of tuberculosis in this case was made on histopathology postoperatively and confirmed by polymerase chain reaction (PCR) on scrapes from the granulomas obtained by microdissection. CONCLUSION: Tuberculosis complicating malignant disease may occur in regions with a high prevalence of disease; with a resurgence of tuberculosis worldwide this association may not be uncommon. The diagnosis and treatment of tuberculosis in a patient with cancer assumes importance as a high mortality has been seen in patients with co-existent disease. PMID- 14564458 TI - Separation of retinoid-induced epidermal and dermal thickening from skin irritation. AB - The ability of the synthetic retinoid MDI-301, in which the carboxylic acid of 9- cis-retinoic acid (9-cis-RA) is replaced with an ester linkage, to induce epidermal and dermal thickening and skin irritation (erythema and flaking) in hairless (rhino) mice following its topical application was investigated in comparison with that of 14-all- trans-retinoic acid (14-all-trans-RA) and 9-cis RA. MDI-301 induced epidermal proliferation leading to a thickened epidermis. Treated animals also demonstrated a prominent band of organized connective tissue immediately below the epidermis. In its ability to induce epidermal thickening, MDI-301 was quantitatively similar to 14-all-trans-RA and 9-cis-RA. However, unlike 14-all-trans-RA and 9-cis-RA, which produced skin irritation associated with a perivascular influx of mononuclear leukocytes into the dermis, there was no evidence of irritation with MDI-301 and little leukocyte infiltration. Intraperitoneal injection of either 14-all-trans-RA or MDI-301 also resulted in epidermal and dermal thickening. Irritation of skin was not observed in these animals but splenomegaly was prominent in animals treated with either agent. PMID- 14564459 TI - Navigation-assisted, minimally invasive implant removal following a triple pelvic osteotomy. AB - INTRODUCTION: Triple pelvic osteotomy is known as a surgical option in young patients suffering from severe hip dysplasia. In most patients, implant removal is performed after bony consolidation. Due to the pelvic anatomy, the conventional technique is often associated with soft-tissue damage and high X-ray exposure. Recent developments in the field of computer-assisted surgery may offer new alternatives. The aim of this case report was to investigate the feasibility of a fluoroscopy-supported navigation system for the implant removal after a triple pelvic osteotomy. MATERIALS AND METHODS: Using the BrainLAB-VectorVision, a fluoroscopy-enhanced navigation system, implants were removed in a 24-year-old female patient 14 months after a triple pelvic osteotomy. RESULTS: The navigation system showed a high feasibility in the instrument visualisation and implant localisation. The screws could be precisely located, simultaneously in different X-ray planes. The total X-ray exposure time was low, totalling 17 s. CONCLUSION: Initial experiences are very promising and show impressively the advantages of fluoroscopy-supported navigation for minimally invasive implant removal. A considerably reduced X-ray exposure for patient and surgeon is possible. The removal of the inserted screw is possible through a stitch incision, with reduced OR trauma and a quicker reconvalescence. PMID- 14564460 TI - Synthetic and tomato-based lycopene have identical bioavailability in humans. AB - BACKGROUND: Bioavailability studies with lycopene have focused on natural sources. A synthetic source has recently become available. AIM OF THE STUDY: To determine the relative bioavailabilities of synthetic and tomato-based lycopene in free living volunteers in a single-blind, randomized, placebo-controlled, parallel trial. METHODS: Three groups (n=12/group) of healthy, normolipemic male and female subjects with a mean baseline serum lycopene concentration of 0.36 micro mol/L took a dose of 15 mg/day total lycopene for 28 days from either Lycovit 10% (beadlets, BASF, Germany) or Lyc-O-Mato (beads, LycoRed Natural Products, Israel) or a placebo (without lycopene) together with the main meal. The increase in serum lycopene from baseline was used as the parameter of bioavailability. RESULTS: Synthetic and tomato-lycopene resulted in significant increases above baseline of serum total lycopene by 0.58 and 0.57 micro mol/L, trans-lycopene by 0.34 and 0.41 micro mol/L, and total-cis-lycopene by 0.24 and 0.16 micro mol/L, whereas no significant changes were found in the placebo treatment. The mean serum total lycopene response to synthetic and natural lycopene was not significantly different. Neither lycopene source affected the other serum carotenoids, viz. alpha-carotene, beta-carotene, beta-cryptoxanthin, zeaxanthin and lutein. CONCLUSION: We conclude that synthetic and natural lycopene are equivalent sources of lycopene and that there is no interaction with other circulating carotenoids. PMID- 14564461 TI - Effects of sulphate- and bicarbonate-rich mineral waters on net and fractional intestinal absorption and urinary excretion of magnesium in rats. AB - Magnesium (Mg) intake is below the recommended daily allowances in many developed countries. Mg-rich mineral waters can provide significant amounts of energy-free Mg and thus help to meet Mg requirements. We assessed the effects of different Mg rich mineral waters on overall intestinal Mg absorption and urinary Mg excretion in 40 rats split into four groups: one received distilled water, another a solution of MgCl(2) and the others two different mineral waters, sulphated water (Hepar) and carbonated water (Badoit) mixed with the diet and as drinking water, for four weeks. The rats were given 3 mg of (26)Mg orally and 0.5 mg of (25)Mg intravenously. They were placed in metabolic cages, and diet consumption, and faeces and urine excretion were monitored during the last four days of the experiment. The rats were then sacrificed and blood was sampled. Mg levels in the diet, faeces, urine and biological samples were measured by atomic absorption spectrometry. Mg stable isotope measurements were performed by ICP/MS. Mg-rich mineral waters significantly increased net intestinal absorption of Mg by more than 30%, but the proportions of both apparent and true intestinal absorption of Mg were similar in all four groups. Thus, net and fractional retention of Mg were similar in the three Mg-supplemented groups. In conclusion, both types of Mg-rich mineral waters studied similarly increased both absorption and urinary excretion of Mg with no positive effect on the overall retention of Mg, probably because the Mg status of the rats was already satisfactory. PMID- 14564462 TI - Iron status of adults in the capital area of Finland. AB - BACKGROUND: Enrichment of wheat flour with iron, which commenced in Finland in the mid-1970s, ceased in 1994. No data on iron status among adults have been published since 1993. AIM OF THE STUDY: To assess the iron status, adequacy of iron intake and dietary sources of iron in Finnish adults. METHODS: A random sample of adults aged 25-64 years in the capital area of Finland was stratified for sex and 10-year age groups. Food consumption was measured with 24-h recall and a 38-item food frequency questionnaire. Iron status was evaluated by haemoglobin (Hb) concentration and other haematological variables from venous blood samples in both genders (n = 239 men and 190 women), and serum ferritin (SF) for women (n = 137). RESULTS: The mean Hb concentration was 133+/-12 g/l and 137 +/- 10 g/l for women aged < 50 years and >/= 50 years, respectively, and 150 +/- 10 g/l for men. In younger women, the mean SF level was 32 +/- 30 micro g/l and 20% of women showed iron depletion (SF < 12 micro g/l),whereas in older women, the respective results were 62 +/- 59 micro g/l and 11 %. The prevalence of anaemia was 5.8 % for women (Hb < 120 g/l) and 1.3 % for men (Hb < 130 g/l). The respective mean intakes of iron were 10 mg/d and 13 mg/d. Cereals and meat were the major dietary sources of iron. CONCLUSIONS: While iron status is fairly good among Finnish males, especially in younger women it is suboptimal,with iron intake failing to reach recommended levels. Food consumption was poorly associated with iron status. PMID- 14564464 TI - Sexual function and continence after ileo pouch anal anastomosis: a comparison between a meta-analysis and a questionnaire survey. AB - BACKGROUND AND AIMS: Ileo pouch anal anastomosis (IPAA) is the standard restorative procedure for patients with ulcerative colitis and familial adenomatous polyposis, but its pros and cons have not been explored in depth. This study analyzed the long-term complications such as incontinence and sexual dysfunction after IPAA. PATIENTS AND METHODS: Netherlands Society for Crohn's disease and Ulcerative Colitis) A questionnaire on complications, sexual dysfunction, and continence was sent to all 137 members of the Netherlands Society for Crohn's disease and Ulcerative Colitis who had IPAA; the questionnaire was returned by 111. Results of the questionnaire were compared with those of a meta-analysis on pooled incidences of complications after IPAA, as previously performed and reported. RESULTS: Pelvic sepsis was reported by 15.3% of respondents. The reported incidence of sexual dysfunction (19.8%), passive incontinence (23.4%), and soiling (39.3%) was significantly higher than that in the meta-analysis. Nevertheless, 90% of the population was satisfied with the results of the IPAA. CONCLUSION: This study underlines that the operation itself, freeing patients of their disease, provides the major satisfaction and improvement of quality of life, even when patients have pouch-related complications such as sexual dysfunction and some degree of fecal incontinence. PMID- 14564465 TI - Bacteria ascend to liver from the bilioenteric conduit after choledochojejunostomy in the cholestatic rat. AB - The high incidence of postoperative cholangitis in children with clinical restoration of bile flow after Roux-Y choledochojejunostomy (RYCJ) assumed the concept of a direct ascending cholangitis caused by pathogens in the intestine, into the intrahepatic bile duct via the porta hepatis. It is also well known that jaundiced animals (patients) are more susceptible to infections of the bile ducts following the procedure of bilioenteric anastomosis. An animal experiment was conducted to compare quantitative bacterial cultures of the choledochojejunostomy area and the liver 24 hours after Escherichia coli (ATCC 25922) or sterile normal saline was injected into the bilioenteric conduit (BEC), following RYCJ in rats with or without the proceeding bile duct ligation. A significant increase of E. coli of the same strain (ATCC 25922), that we injected into the BEC, was proved with pulse-field gel electrophoresis (PFGE) and shown in the liver of the jaundiced rats receiving E. coli (ATCC 25922), compared to that in the nonjaundiced rats with normal saline treatment. It is concluded that bacteria often ascend early to the liver from the BEC following RYCJ. This ascending cholangitis model might be produced for further studies. PMID- 14564466 TI - Blunt renal injuries in Turkish children: a review of 205 cases. AB - A retrospective analysis of the medical records of 205 children with renal injuries secondary to blunt abdominal trauma is used to make recommendations regarding the initial diagnostic and therapeutic approaches in this type of patient. It was found that the absence of hematuria on initial urinalysis does not exclude a serious renal injury. Thus, following blunt abdominal trauma, all children should undergo imaging procedures to exclude renal injury, whether they have hematuria or not. Ultrasound is a good initial screening procedure in all patients. Computed tomography is recommended for the definitive evaluation of suspected major renal injuries. Since even major renal injuries may heal without surgical intervention, conservative management is the recommended initial treatment of choice. Surgery is reserved for those children who are hemodynamically unstable and those that develop complications. PMID- 14564467 TI - A phylogenetic analysis of basal metabolism, total evaporative water loss, and life-history among foxes from desert and mesic regions. AB - We measured basal metabolic rate (BMR) and total evaporative water loss (TEWL) of species of foxes that exist on the Arabian Peninsula, Blanford's fox (Vulpes cana) and two subspecies of Red fox (Vulpes vulpes). Combining these data with that on other canids from the literature, we searched for specialization of physiological traits among desert foxes using both conventional least squares regression and regressions based on phylogenetic independent contrasts. Further, we explored the consequences of reduced body size of foxes on life history parameters such as litter size and neonate mass. For Blanford's foxes, Red foxes from the central desert of Arabia, and Red foxes from the more mesic Asir mountains, body mass averaged 1,285 +/- 52 g, 1,967 +/- 289 g, and 3,060 +/- 482 g, respectively, whereas mean BMR, during summer, was 304.5 +/- 32.3 kJ/day, 418.0 +/- 32.4 kJ/day, and 724.1 +/- 120.2 kJ/day (+/- SD). An analysis of covariance with body mass as a covariate showed no statistical differences in BMR among foxes. Analysis of covariance indicated that Red fox from the Asir mountains had a higher TEWL than Red foxes from central Arabia or than Blanford's foxes also from the mountains. Comparisons of all species of desert and mesic foxes showed no significant differences in BMR, nor did desert foxes have a significantly lower BMR than other carnivores. TEWL of desert foxes was lower than other more mesic carnivores; deviations in TEWL ranged from -17.7% for the Fennec fox (Fennecus zerda) to -57.4% for the Kit fox (Vulpes velox). Although desert foxes have a BMR comparable to other more mesic species, it appears that desert foxes do have a smaller body mass, lowering overall energy requirements. We attribute this reduction in body size to the "resource limitation hypothesis" whereby natural selection favors smaller individuals in a resource-limited environment, especially during periods of severe food shortage. However, until common garden experiments are performed, developmental plasticity and acclimation cannot be ruled out as contributors to this pattern. PMID- 14564468 TI - Can two streams of auditory information be processed simultaneously? Evidence from the gleaning bat Antrozous pallidus. AB - A tenet of auditory scene analysis is that we can fully process only one stream of auditory information at a time. We tested this assumption in a gleaning bat, the pallid bat (Antrozous pallidus) because this bat uses echolocation for general orientation, and relies heavily on prey-generated sounds to detect and locate its prey. It may therefore encounter situations in which the echolocation and passive listening streams temporally overlap. Pallid bats were trained to a dual task in which they had to negotiate a wire array, using echolocation, and land on one of 15 speakers emitting a brief noise burst in order to obtain a food reward. They were forced to process both streams within a narrow 300 to 500 ms time window by having the noise burst triggered by the bats' initial echolocation pulses as it approached the wire array. Relative to single task controls, echolocation and passive sound localization performance was slightly, but significantly, degraded. The bats also increased echolocation interpulse intervals during the dual task, as though attempting to reduce temporal overlap between the signals. These results suggest that the bats, like humans, have difficulty in processing more than one stream of information at a time. PMID- 14564469 TI - Comparison of visual grading analysis and determination of detective quantum efficiency for evaluating system performance in digital chest radiography. AB - A study was conducted to compare physical and clinical system performance in digital chest radiography. Four digital X-ray modalities, two storage-phosphor based systems and two generations of a CCD-based system, were evaluated in terms of both their imaging properties (determination of presampling MTF and DQE) and clinical image quality (grading of the reproduction of anatomical details of 23 healthy volunteers using both absolute and relative visual grading analysis). One of the two storage-phosphor systems performed best in both evaluations and the first generation of the CCD-based system was rated worst; however, the other two systems were ranked differently with the two methods. The newest CCD-based system yielded a higher clinical image quality than the second storage-phosphor system, although the latter presented a DQE substantially higher than the former. The results show that clinical performance cannot be predicted from determinations of DQE alone, and that a system with lower DQE, under the quantum-saturated conditions in chest radiography, can outperform a system with higher DQE if the image processing used on the former is more effective in presenting the information in the image to the radiologist. PMID- 14564470 TI - Radiofrequency thermal ablation with expandable needle of focal liver malignancies: complication report. AB - The aim of this study was to describe type and rate of complications in a series of patients with liver tumors treated by the radiofrequency (RF) expandable system. A total of 166 patients, 114 with hepatocellular carcinoma (HCC; 92 small HCC, 22 large) and 52 with liver metastasis, were treated by the percutaneous RF expandable system. In large HCCs, RF ablation was performed after tumor ischemia (TAE or balloon stop flow of the hepatic artery). Major complications were those that delayed hospital discharge, with or without additional medical procedures or treatments. Minor complications did not require an additional hospital stay. No deaths occurred. Among 151 patients followed, there were 7 (4.6%) early major complications-severe pain with session interruption in 3 cases, capsular necrosis in 1 case, 1 abdominal wall necrosis, 1 dorsal burning, 1 peritoneal hemorrhage and 3 (1.9%) delayed major complications: sterile fluid collection at the site of the treated tumor in 2 cases and cutaneous seeding in 1 case. There were 49 (32.5%) minor complications. The complication rate is similar to that observed after percutaneous alcohol injection (PEI). With the cooled system, the complication rate is seemingly lower but that may well be due to a different definition of major complications. The seeding rate after expandable system ablation is lower than after PEI. It is the same as or lower than that in other series of patients treated by the cooled system. PMID- 14564472 TI - Hepatic calcification following umbilical vein catheterization in a premature baby. PMID- 14564471 TI - (3)He-MRI in follow-up of lung transplant recipients. AB - The aim of this study was to evaluate the possible contribution of (3)He-MRI to detect obliterative bronchiolitis (OB) in the follow-up of lung transplant recipients. Nine single- and double-lung transplanted patients were studied by an initial and a follow-up (3)He-MRI study. Images were evaluated subjectively by estimation of ventilation defect area and quantitatively by individually adapted threshold segmentation and subsequent calculation of ventilated lung volume. Bronchiolitis obliterans syndrome (BOS) was diagnosed using pulmonary function tests. At (3)He-MRI, OB was suspected if ventilated lung volume had decreased by 10% or more at the follow-up MRI study compared with the initial study. General accordance between pulmonary function testing and (3)He-MRI was good, although subjective evaluation of (3)He-MRI underestimated improvement in ventilation as obtained by pulmonary function tests. The (3)He-MRI indicated OB in 6 cases. According to pulmonary function tests, BOS was diagnosed in 5 cases. All diagnoses of BOS were also detected by (3)He-MRI. In 2 of these 5 cases, (3)He MRI indicated OB earlier than pulmonary function tests. The results support the hypothesis that (3)He-MRI may be sensitive for early detection of OB and emphasize the need for larger prospective follow-up studies. PMID- 14564473 TI - Classification and imaging of vascular malformations in children. AB - Accurate diagnosis and classification of haemangiomas and vascular malformations is essential for prediction of outcome and rational therapeutic intervention. The purpose of this review is to demonstrate how radiological techniques, including projectional radiography, ultrasound and colour flow imaging, magnetic resonance imaging, and conventional venography and arteriography can aid in the diagnosis and classification of vascular malformations in children, and to guide the radiologist in the use of modern nomenclature. PMID- 14564474 TI - Covered stenting in patients with lifting of gastric and high esophago-tracheal fistula. PMID- 14564475 TI - Self-fertile apple resulting from S-RNase gene silencing. AB - Self-incompatibility (SI) restricts fertilisation and fruit setting in many tree fruit crops. In apple, we have produced transgenic trees harbouring extra copies of the endogenous S-gene controlling SI. Two independent transgenic genotypes were characterised in detail. Controlled self- and cross-pollination of the flowers of trees from both genotypes over a 3-year-period showed that the transgenic lines produced normal levels of fruit and seeds after selfing. In contrast, the controls produced much less fruit following self- compared to cross pollination. Fruit set data correlated with the results of microscopic evaluation of pollen tube growth through the pistil, which revealed inhibition after selfing in the controls but not in the transgenic lines. The self-fertile phenotype was associated with the complete absence of pistil S-RNase proteins, which are the products of the targeted S-gene. These results confirm that self-fertility was due to inhibition of expression of the S-RNase gene in the pistil, resulting in un-arrested self-pollen tube growth, and fertilisation. PMID- 14564477 TI - Amelioration of doxorubicin-induced cardiac and renal toxicity by pirfenidone in rats. AB - PURPOSE: Doxorubicin (DXR) is an anthracycline glycoside with a broad spectrum of therapeutic activity against various tumors. However, the clinical use of DXR has been limited by its undesirable systemic toxicity, especially in the heart and kidney. This study was designed to test the effectiveness of dietary intake of pirfenidone (PD) against DXR-induced cardiac and renal toxicity. METHODS: Male Sprague Dawley rats were placed into four treatment groups: saline injected intraperitoneally (i.p.) plus regular diet (SA+RD); DXR i.p. plus regular diet (DXR+RD); saline i.p. plus the same diet mixed with 0.6% PD (SA+PD); and DXR i.p. plus the same diet mixed with 0.6% PD (DXR+PD). The animals were fed regular or regular plus PD diets 3 days prior to i.p. injections of either saline or DXR and continuing throughout the study. A total dose of DXR (16.25 mg/kg) or an equivalent volume of saline was administered in seven injections (2.32 mg/kg per injection) three times per week with an additional dose on the 12th day. At 25 days following the last DXR or saline injection, some animals were anesthetized for the measurement of cardiac and pulmonary function, and others were killed by an overdose of pentobarbital. At the time the animals were killed, abdominal fluid was collected. Kidney and heart were removed, weighed, fixed with 10% formalin or frozen in liquid nitrogen. The fixed tissues were used for histological examination and the frozen tissues were used for biochemical studies. RESULTS: The average volumes of abdominal fluid in the DXR+RD and DXR+PD groups were 9.42 ml and 3.42 ml and the protein contents of abdominal fluid in the DXR+RD and DXR+PD groups were 218 mg and 70 mg, respectively. A 12.5% mortality occurred in the DXR+RD group as compared to 0% in DXR+PD group. There were no changes in any of the cardiac or pulmonary physiological parameters in any of the four groups. The changes in the heart and kidney of the DXR+RD group included reduction in organ weight, increase in hydroxyproline content of heart, increase in hydroxyproline, and lipid peroxidation in the kidney and plasma, and increase in protein concentration in urine as compared to rats in the control, SA+RD and SA+PD groups. Treatment with PD abrogated the DXR-induced increases in hydroxyproline content in the heart and kidney, lipid peroxidation of the kidney and plasma, and protein content of the urine in the DXR+PD group. DXR treatment alone caused disorganization of cardiac myofibrils, vacuolization of the myofibers, and renal tubular dilation with protein casts in both the cortical and medullary regions. Treatment with PD minimized the DXR-induced histopathological changes of heart and kidney in the DXR+PD group. CONCLUSIONS: Treatment with PD reduced the severity of DXR-induced toxicity as assessed by reduced mortality, diminished volume of recovered fluid in the abdominal cavity, and severity of cardiac and renal lesions at both the biochemical and morphological levels. These results indicate that PD has the potential to prevent DXR-induced cardiac and renal damage in humans on DXR therapy. PMID- 14564476 TI - Chromosome rearrangements in isolates that escape from het-c heterokaryon incompatibility in Neurospora crassa. AB - Chromosomal rearrangement is implicated in human cancers and hereditary diseases. Mechanisms generating chromosomal rearrangements may be shared by a variety of organisms. Spontaneous chromosomal rearrangements, especially large deletions, take place at high frequency in isolates that escape from heterokaryon incompatibility in Neurospora crassa. In this study, chromosomal rearrangements were detected in strains that had escaped from het-c heterokaryon incompatibility in N. crassa. A vc1 mutant carried a 20-kbp deletion covering five ORFs. A vc2 mutant carried a complex chromosome rearrangement with an 8-kbp deletion covering three ORFs, a 34-bp deletion and an 80-kbp inversion. The break-points of chromosome rearrangements in the vc1 and vc2 mutants all have direct repeats of 2 bp, similar to the break-points of some chromosome rearrangements associated with human cancer and genetic diseases. An ahc mutant carried a 31-kbp deletion covering at least 11 ORFs and a het-c deletion mutant carried a 7-kbp deletion covering two ORFs. Additional chromosomal rearrangements occurred in these two strains. These results indicate that escape from heterokaryon incompatibility can be used as a model system for chromosome rearrangement and DNA-repair studies. The impact of the chromosomal rearrangements is discussed, especially the deletion of the predicted ORFs on the phenotype of mutants. PMID- 14564478 TI - Chronic idiopathic myelofibrosis: prognostic impact of myelofibrosis and clinical parameters on event-free survival in 122 patients who presented in prefibrotic and fibrotic stages. A retrospective study identifying subgroups of different prognoses by using the RECPAM method. AB - In chronic idiopathic myelofibrosis (CIMF) the factors predicting survival in patients who were already in the fibrotic stage have been well documented by numerous studies. Prefibrotic stages were only rarely evaluated so that the prognostic impact of myelofibrosis is currently not well known. Also predictive factors for disease-related events were not included in those studies. Thus, we evaluated the prognostic impact of myelofibrosis and other histopathological (megakaryocytes, blasts) and clinical [age, gender, splenomegaly, chemotherapy, hemoglobin (Hb), leukocyte, and platelet count] parameters in 122 patients in fibrotic and prefibrotic stages of CIMF on event-free survival. The statistical analysis was performed using the univariate log-rank test and the multivariate recursive partition and amalgamation (RECPAM) approach. In 62 patients disease related events occurred during a mean observation period of 58 months. In univariate analysis they were associated with blast increase in the bone marrow. In RECPAM analysis a shorter event-free survival was found in anemic patients (mean: 9.3 months). In nonanemic patients older than 60 years, advanced myelofibrosis was associated with a shorter event-free mean survival of 23.2 months versus 69.3 months in less advanced cases. A slight or moderate myelofibrosis was not found to have a prognostic impact on event-free survival. The longest event-free survival was found in nonanemic patients who were younger than 60 years (mean: 185 months), regardless of the grade of myelofibrosis. Thus, we found that the most relevant prognostic parameter for event-free survival in CIMF were the Hb value, age, and grade of myelofibrosis. PMID- 14564479 TI - The discomallear ligament and the anterior ligament of malleus: an anatomic study in human adults and fetuses. AB - According to some reports, movement of the malleus, resulting from anterior hypertension on the discomallear ligament (DML), could produce aural symptoms related with damage to middle ear structures. The aim of this study was to examine the topographic relationship of the DML and the anterior ligament of malleus (ALM). Four fetuses and 16 adult hemi-sectioned heads were used to determine the anatomic-clinical relevance of DML and ALM in temporomandibular disorder. In fetal specimens, the DML was distinctly interposed between the malleus and the disc of the temporomandibular joint (TMJ), and the ALM had a structure apparently composed of the superior and inferior lamellae, running anteriorly in continuation with the sphenomandibular ligament (SML) through the future petrotympanic fissure (PTF). In all adult specimens, the DML was inserted into the malleus, and it expanded broadly toward the disc and capsular region of the TMJ in a triangular shape and inserted into the disc and capsule of the TMJ. The two-lamellae structure of the ALM was not distinguishable in adult specimens. The overstretched ALM resulted in movement of the malleus in five cases, but similar tension applied to the DML did not cause any movement of the malleus. This result provides an indication of the clinical significance of the ALM, a ligamentous structure continuous with the SML. It is apparent that the ALM has the potential to cause aural symptoms as a result of damage to the middle ear structure. PMID- 14564480 TI - Concurrent aberrant right gastric vein directly draining into the liver and variations of the hepatic artery. AB - We report an autopsy on a 46-year-old man, a case that presented the concurrence of two rare vascular variations of the lesser omentum: aberrant right gastric vein draining directly into the liver, and multiple hepatic arteries. Although the left gastric vein emptied into the left aspect of the portal vein, the right one was found to ascend from the gastric lesser curvature along the right aspect of the common bile duct and to reach directly the porta hepatis. A left hepatic artery originating from the left gastric artery entered the porta hepatis in conjunction with the left ramus of the portal vein. A predominant right hepatic artery arose from the superior mesenteric artery and entered the porta hepatis in conjunction with the right ramus of the portal vein. The proper hepatic artery originating from the celiac artery entered the porta hepatis in conjunction with the aberrant right gastric vein. The possibility of a common underlying mechanism for these rare vascular variations is discussed. PMID- 14564482 TI - Identification of new MHC-restriction elements for presentation of the p210(BCR ABL) fusion region to human cytotoxic T lymphocytes. AB - Chronic myelogenous leukemia (CML) is characterized by a t(9;22) translocation resulting in expression of BCR-ABL fusion oncoproteins which are unique to the leukemic cells, necessary for oncogenesis, and potentially immunogenic. We have previously shown that human dendritic cells transduced with an adeno-associated virus vector encoding the fusion region of the b3a2 splice variant (p210(b3a2)) of the BCR-ABL oncoprotein elicit specific T-cell responses in vitro. Two cytotoxic T lymphocyte (CTL) clones generated in this fashion displayed restriction with previously unreported HLA alleles. The first, T1/B9, was CD4(+) and restricted by DRB5*0101 (autologous) or DRB1*1101 (allogeneic). The minimum cytotoxic epitope (MCE) binding to DRB5*0101 for this clone was identified as FKQSSKALQ, overlapping the p210(b3a2) fusion point (boldface). The MCE of DRB1*1101 for this clone differed from DRB5*0101, but also included the fusion point. The clonality of CTL T1/B9 was verified by analyses of TCRalpha/beta chain usage and DNA sequence analyses. To our knowledge, this is the first description of a single clone recognizing both DRB5*0101 and DRB1*1101. The other CTL clone, T1/33, was CD8+ and recognized HLA-B*3501 or B*3503 complexed with an MCE, RPVASDFEP, derived from the c-abl sequence in proximity to the p210(b3a2) fusion point. K562 cells transfected with plasmids encoding HLA-DRA + B5*0101, B*3501, or B*3503 but not controls expressing DRA + DRB1*1501 were lysed by cognate CTL clones, confirming that DRB5*0101 and B*3501/3 could present p210(b3a2) joining region epitopes via endogenous processing. The identification of three additional HLA alleles (DRB5*0101, B*3501, and B*3503) presenting the p210(b3a2) fusion region antigen will broaden the application of vaccine strategies for targeting CML cells. The findings of single CTL clones cross-recognizing autologous (DRB5*0101 or B*3501) and allogeneic (DRB1*1101 or B*3503) HLA alleles presenting BCR-ABL fusion-region epitopes implies the potential separation of graft-versus leukemia from graft-versus-host effects. PMID- 14564483 TI - Phase I pilot clinical trial of human IgM monoclonal antibody to ganglioside GM3 in patients with metastatic melanoma. AB - PURPOSE: A human monoclonal antibody (L612 HuMAb) that binds to ganglioside GM3 has been developed in our laboratory. L612 HuMAb is a 100% human IgM protein. L612 HuMAb binds to cell surface of melanoma and can kill the cells in the presence of complement. The primary objective of this study was to test the toxicity and pharmacokinetics associated with administration of L612 HuMAb to melanoma patients whose tumor cells expressed GM3. EXPERIMENTAL DESIGN: Nine patients with measurable metastatic melanoma (American Joint Committee on Cancer stage IV) were entered in the study. Eight had failed previous treatments that included chemotherapy, radiation therapy, melanoma cell vaccine, and/or biological therapy. All patients received a 48-h continuous infusion of L612 HuMAb at a dose of 960 mg, 1,440 mg, or 1,920 mg. Five of these patients received a second infusion and one patient received a third infusion, all with the previous dose. RESULTS: Toxicity was limited to transient and mild pruritus and skin rash. One patient complained of pain at the site of subcutaneous metastases. Serum antibody levels peaked 24 to 48 h after starting the infusion. Two patients, one receiving a single course of 960 mg (612 mg/m(2)) and the second receiving two courses of 1,440 mg (911 mg/m(2)) followed by surgical therapy, are without evidence of disease >5 years after antibody infusion. CONCLUSIONS: The human IgM monoclonal antibody, L612 HuMAb, was well tolerated. Infusion of L612 HuMAb appears to produce significant antitumor activity in melanoma patients. PMID- 14564485 TI - Conversion of adamsite (phenarsarzin chloride) by fungal manganese peroxidase. AB - Fungal manganese peroxidase was found to convert the persistent chemical warfare agent adamsite (phenarsarzin chloride) in a cell-free reaction mixture containing sodium malonate, Mn(2+) ions, and reduced glutathione. The organo-arsenical compound disappeared completely within 48 h accompanied by the formation of a more polar metabolite with a clearly modified UV spectrum. Thus, As(III) in the adamsite molecule was oxidized by manganese peroxidase to As(V) which added dioxygen and released chloride. PMID- 14564484 TI - Cystic synovial sarcomas: imaging features with clinical and histopathologic correlation. AB - OBJECTIVE: To characterize the radiological and clinicopathologic features of cystic synovial sarcoma. DESIGN AND PATIENTS: Seven patients with primary cystic synovial sarcoma were evaluated. Computed tomography (CT) and magnetic resonance (MR) imaging were undertaken at the first presentation. The diagnosis of synovial sarcoma was made on the basis of histological examinations followed by molecular analysis. Radiological and clinicopathologic findings were reviewed. RESULTS: CT showed well-defined soft tissue mass without cortical bone erosion and invasion. Calcification was seen at the periphery of the mass in three cases. T2-weighted MR images showed multilocular inhomogeneous intensity mass in all cases, five of which showed fluid-fluid levels. On gross appearance, old and/or fresh hematomas were detected in six cases. In the one remaining case, microscopic hemorrhage in the cystic lumen was proven. Four cases had poorly differentiated areas. In five cases prominent hemangiopericytomatous vasculature was observed. Histologic grade was intermediate in one tumor and high in six. One case had a history of misdiagnosis for tarsal tunnel syndrome, one for lymphadenopathy, two for sciatica and two for hematoma. CONCLUSION: All cystic synovial sarcomas demonstrated multilocularity with well-circumscribed walls and internal septae. Synovial sarcoma should be taken into consideration in patients with deeply situated multicystic mass with triple signal intensity on T2-weighted MR imaging. PMID- 14564487 TI - Copper dissociation as a mechanism of fungal laccase denaturation by humic acid. AB - Effects of humic acid on removal of hydroxy polychlorobiphenyls (PCBs) with laccase from Trametes versicolor were studied. In the absence of humic acid, hydroxy PCBs were rapidly degraded by laccase. However, the rate constants decreased with increasing humic acid concentration, the reactions being completely inhibited at 150 mg l(-1) of humic acid. Peroxidase from Arthromyces ramosus was not inhibited by the same treatment. The activity of humic acid deactivated laccase was completely restored by copper ions (500 microM of Cu2+ in 150 mg l(-1) of humic acid), but not by other metal ions (Zn2+, Fe2+ and Hg2+). Humic acid-deactivated laccase purified by gel permeation chromatography (GPC) showed no activity against 2,2'-azino-bis(3-ethylbenzthiazoline sulfonic acid) diammonium salt and 3,5-dichloro-4-hydroxybiphenyl, but its activity was restored by copper ion treatment. Humic acid-deactivated laccase showed similar properties, such as GPC retention time and copper ion requirements for activity, to those of laccase deactivated by nitrilotriacetic acid. The extent of humic acid inhibition, expressed as activity non-recoverable by copper ion treatment, increased over time more rapidly than that of the humic acid-free control. These results suggest that short-term inactivation of laccase, i.e., less than 1 day, is attributable to a depletion of copper ion. PMID- 14564486 TI - Biotransformation of glucose to 5-keto-D-gluconic acid by recombinant Gluconobacter oxydans DSM 2343. AB - For the conversion of glucose to 5-keto-D-gluconate (5-KGA), a precursor of the industrially important L-(+)-tartaric acid, Gluconobacter strains were genetically engineered. In order to increase 5-KGA formation, a plasmid-encoded copy of the gene encoding the gluconate:NADP-5 oxidoreductase (gno) was overexpressed in G. oxydans strain DSM 2434. This enzyme is involved in the nonphosphorylative ketogenic oxidation of glucose and oxidizes gluconate to 5 KGA. As the 5-KGA reductase activity depends on the cofactor NADP+, the sthA gene (encoding Escherichia coli transhydrogenase) was cloned and overexpressed in the GNO-overproducing G. oxydans strain. Growth of the sthA-carrying strains was indistinguishable from the G. oxydans wild-type strain and therefore they were chosen for the coupled overexpression of sthA and gno. G. oxydans strain DSM 2343/pRS201-gno-sthA overproducing both enzymes showed an enhanced accumulation of 5-KGA. PMID- 14564489 TI - The biosynthesis of glycopeptide antibiotics--a model for complex, non ribosomally synthesized, peptidic secondary metabolites. AB - Glycopeptide antibiotics are a class of widely known natural compounds produced by Actinomycetes. Vancomycin, the first member of the glycopeptide family to be discovered, was described in 1955 and used as an antibiotic soon thereafter. During the past 50 years numerous contributions on the structure, mode of action, and therapeutic features of vancomycin have been published. Recently, there has been considerable progress in elucidating the biosynthesis of glycopeptide antibiotics by combining molecular biology and analytical chemistry methods. Here, we provide an overview of the current knowledge regarding biosynthetic glycopeptide assembly. PMID- 14564488 TI - Evaluation of different organic phases for water-in-oil xanthan fermentation. AB - Water-in-oil (W/O) fermentation technology has the potential for overcoming the problems related with high broth viscosity in xanthan fermentations. By dispersing the aqueous broth in a continuous organic phase, the broth-thickening mechanisms are confined within the aqueous droplets without significantly increasing the overall viscosity. In this study, xanthan fermentations were made with perfluorocarbon (PFC) or vegetable oil as the organic phase. The results were compared with those obtained previously using n-hexadecane as the organic phase, to evaluate the effects of various properties. PFC provided easy phase separation at the end of fermentation but required higher power input for agitation, a direct result of its high density. The aqueous droplets formed were large (400-450 microm), limiting the cell concentration employable due to the occurrence of oxygen starvation in the inner core. One main advantage of using vegetable oil was its low cost. In addition, vegetable oil provided much finer droplets (<120 microm) and produced high xanthan concentrations (>100 g l(-1)). However, complete phase separation for product recovery was difficult to achieve. Fermentations in both organic phases were terminated by the occurrence of phase inversion to highly viscous O/W dispersions at aqueous-phase volume fractions of 0.53-0.56. The initial fraction was 0.3 but changed due to base addition for pH adjustment and nutrient addition for prolonged production. PMID- 14564490 TI - Utilization of the buffering capacity of corn steep liquor in bacterial cellulose production by Acetobacter xylinum. AB - Acetobacter xylinum BPR2001 produces water-insoluble bacterial cellulose (BC). Using a pH sensor for the accurate control of pH, which is one of the most critical factors for efficient BC production, is difficult especially in a baffled shake-flask and an airlift reactor. The buffering capacity of corn steep liquor (CSL) was estimated by measuring beta (buffering capacity) values in advance and was used to maintain the pH within the optimal range during the production of BC. When CSL was added to either a shake-flask, a stirred-tank reactor or an airlift reactor, BC production was almost the same as that in cultivations where pH was controlled manually or by a pH sensor. PMID- 14564491 TI - Expression of foreign proteins in Escherichia coli by fusing with an archaeal FK506 binding protein. AB - Improper protein-folding often results in inclusion-body formation in a protein expression system using Escherichia coli. To express such proteins in the soluble fraction of E. coli cytoplasm, we developed an expression system by fusing the target protein with an archaeal FK506 binding protein (FKBP). It has been reported that an archaeal FKBP from a hyperthermophilic archaeon, Thermococcus sp. KS-1 (TcFKBP18), possesses not only peptidyl-prolyl cis-trans isomerase activity, but also chaperone-like activity to enhance the refolding yield of an unfolded protein by suppressing irreversible protein aggregation. To study the effect of this fusion strategy with FKBP on the expression of foreign protein in E. coli, a putative rhodanese (thiosulfate sulfurtransferase) from a hyperthermophilic archaeon and two mouse antibody fragments were used as model target proteins. When they were expressed alone in E. coli, they formed insoluble aggregates. Their genes were designed to be expressed as a fusion protein by connecting them to the C-terminal end of TcFKBP18 with an oligopeptide containing a thrombin cleavage site. By fusing TcFKBP18, the expression of the target protein in the soluble fraction was significantly increased. The percentage of the soluble form in the expressed protein reached 10-28% of the host soluble proteins. After purification and protease digestion of the expressed antibody fragment-TcFKBP18 fusion protein, the cleaved antibody fragment (single-chain Fv) showed specific binding to the antigen in ELISA. This indicated that the expressed antibody fragment properly folded to the active form. PMID- 14564493 TI - Support groups for cancer patients. AB - Within the last two decades psychosocial group interventions have been developed to help cancer patients cope better with the psychosocial sequelae of cancer diagnosis and treatment. Support groups include a variety of different approaches some of which focus on behavioral aspects and symptoms (e.g. pain, fatigue) and some on the expression of emotions. Most of these support programs are structured and short-term and include elements such as delivery of information, emotional and social support, stress management strategies based on the cognitive behavioral approach and the teaching of relaxation techniques. Beyond individual therapy, group therapies can address cancer-related issues to enable patients to gain emotional support from other patients with similar experiences and to use these experiences to buffer the fear of dying and the unknown future. One of the overall therapeutic targets is the promotion of the patient's individual resources. Therefore, such groups are helpful not only for the patients, but also for their spouses and other family members, in relieving the cancer-related distress. In Germany, support groups are established in rehabilitation clinics as well as outpatient programs and play an important role in palliative and supportive care of cancer patients. Against the background of changes in the patients' role, the increasing availability of information technology (e.g. the internet) and patient advocacy in cancer treatment, support groups may be understood as a mean of empowerment of the patient. The need for group interventions such as outpatient programs for cancer patients is claimed not only by the health professionals but also by the patients themselves. There is some research emphasizing that avoidance of feelings, denial of concerns, feelings of helplessness and social isolation are correlated with poorer health outcome and poorer quality of life. Many empirical studies have provided evidence-based knowledge that structured group interventions for cancer patients improve psychological wellbeing, reduce anxiety and depression, and improve quality of life, coping and mental adjustment. Positive effects on survival have even been reported, but these effects have not yet been proven. PMID- 14564492 TI - Characterization of the horse (Equus caballus) IGHA gene. AB - Nucleotide sequences of the immunoglobulin constant heavy chain genes of the horse have been described for IGHM, IGHG and IGHE genes, but not for IGHA. Here, we provide the nucleotide sequence of the genomic IGHA gene of the horse ( Equus caballus), including its secretion region and the transmembrane exon. The equine IGHA gene shows the typical structure of a mammalian IGHA gene, with only three exons, separated by two introns of similar size. The hinge exon is located at the 5' end of the CH2 exon and encodes a hinge region of 11 amino acids, which contains five proline residues. The coding nucleotide sequence of the secreted form of the equine IGHA gene shares around 72% identity with the human IGHA1 and IGHA2 genes, as well as the bovine, ovine, porcine and canine IGHA genes, without distinct preference for any of these species. The same species also cluster together in a phylogenetic tree of the IGHA coding regions of various mammals, whereas rodent, rabbit, marsupial and monotreme IGHA genes each build a separate cluster. PMID- 14564494 TI - The importance of clinical practice guidelines (CPGs) for the quality and development of supportive care in Central and Eastern European (CEE) countries. AB - Supportive care CPGs are intended to improve the quality of supportive care received by cancer patients. They hold potential benefits for health-care professionals, health-care organizations and patients. In order to provide an effective influence of CPGs on supportive care development in CEE countries their validity and successful implementation in clinical practice must be ensured. This would involve several important steps: (1) achievement of a consensus across Europe about what constitutes supportive care in cancer; (2) identification of supportive care priorities in CEE countries to be covered by the guidelines; and (3) searching existing CPGs relevant to previously identified priorities. Valid international CPGs could then be translated and adapted to local resources and circumstances in CEE countries and used to prepare national supportive care CPGs which could assist in the development of national supportive care standards. Skills, expertise and additional investment are required for local adaptation, dissemination and implementation of international supportive care CPGs. PMID- 14564495 TI - The timing of bud burst and its effect on tree growth. AB - A phenology model for estimating the timings of bud burst--one of the most influential phenological phases for the simulation of tree growth--is presented in this study. The model calculates the timings of the leafing of beech (Fagus sylvatica L.) and oak (Quercus robur L.) and the May shoot of Norway spruce (Picea abies L.) and Scots pine (Pinus sylvestris L.) on the basis of the daily maximum temperature. The data for parameterisation and validation of the model have been taken from 40 climate and 120 phenological stations in southern Germany with time series for temperature and bud burst of up to 30 years. The validation of the phenology module by means of an independent data set showed correlation coefficients for comparisons between observed and simulated values of 54% (beech), 55% (oak), 59% (spruce) and 56% (pine) with mean absolute errors varying from 4.4 days (spruce) to 5.0 days (pine). These results correspond well with the results of other--often more complex--phenology models. After the phenology module had been implemented in the tree-growth model BALANCE, the growth of a mixed forest stand with the former static and the new dynamic timings for the bud burst was simulated. The results of the two simulation runs showed that phenology has to be taken into account when simulating forest growth, particularly in mixed stands. PMID- 14564496 TI - Postpartum hemolytic uremic syndrome in a 17-year-old Filipina primigravid. AB - A 17-year-old Filipina primigravid developed acute renal failure secondary to hemolytic uremic syndrome (HUS) after undergoing emergency cesarean section for severe pre-eclampsia and abruptio placenta. She underwent hemodialysis with concurrent infusions of fresh-frozen plasma and packed red cells for 5 weeks. Renal biopsy revealed findings consistent with HUS with glomerular crescents. She received three doses of pulse methylprednisolone followed by oral prednisone. Renal function improved 5 weeks after the onset of HUS. The pathogenesis, differential diagnosis, and treatment options of postpartum HUS are discussed. PMID- 14564497 TI - Lack of bilirubin removal during dialysis without an impact on creatinine clearance. PMID- 14564498 TI - Mesenteric thrombosis causing short bowel syndrome in nephrotic syndrome. AB - Nephrotic patients are at risk of developing venous and arterial thrombotic complications. Pulmonary embolism due to affected deep leg veins is by far the most common event. Renal or cerebral vein thromboses have been described. Thrombosis of arterial vessels is less frequent. Mesenteric infarction is a rare but severe complication in patients with nephrotic syndrome (NS). We report a 7 year-old boy with a steroid-dependent (SD) NS and a homozygous mutation of methylenetetrahydrofolate reductase, increasing the risk of thromboembolic events. He developed a thrombosis of his superior mesenteric artery during his ninth relapse, which was responsible for a necrosis of 240 cm of his small bowel, necessitating resection of necrotic parts and double external ostomy diversion. Remission was achieved with pulse prednisolone therapy. Corticoids were reduced over 4 months progressively. Oral cyclosporin A (CyA) was initiated for long-term treatment. Due to a short bowel syndrome with severe malabsorption, even oral administration of 22.5 mg/kg per day CyA did not lead to sufficient plasma levels. Intravenous cyclophosphamide pulse therapy over 6 months led to a complete remission. No relapse occurred over a period of more than 5 months after the last cyclophosphamide pulse. Anticoagulation and screening for increased susceptibility for thrombotic events are necessary in every nephrotic patient. Intravenous cyclophosphamide pulse therapy is a useful alternative in SDNS with impaired intestinal absorption of applied immunosuppressive drugs. PMID- 14564499 TI - Biological waste gas treatment with a modified rotating biological contactor. Iota. Control of biofilm growth and long-term performance. AB - In this work, we introduce a modified rotating biological contactor (RBC) system and demonstrate its feasibility by applying the newly devised process to the biological treatment of artificial waste gas. In the proposed system, the waste gas is introduced to the bioreactor in the spacings between the rotating discs through a hollow shaft, thus allowing for intimate gas-liquid contact. A 91-l modified RBC containing 20 biofilm support discs 40 cm in diameter was used in the experiments. Toluene was used as the model pollutant, and the system was operated under standard operating conditions for more than one year in order to investigate its long-term performance and assess its ability to control the growth of the biofilm. It was demonstrated that the proposed system allows to efficiently control the growth of the biofilm, thus overcoming the clogging problem inherent in most conventional methods for the biological treatment of waste gas. Moreover, the system was shown to exhibit stationary long-term performance for a period of more than one year, hence indicating its feasibility for industrial application. PMID- 14564500 TI - Differential host use in two highly specialized ant-plant associations: evidence from stable isotopes. AB - Carbon and nitrogen stable isotopes were used to examine variation in ant use of plant resources in the Cecropia obtusifolia / Azteca spp. association in Costa Rica. Tissue of ants, host plants and symbiotic pseudococcids were collected along three elevation transects on the Pacific slope of Costa Rica's Cordillera Central, and were analyzed for carbon and nitrogen isotopic composition. Worker carbon and nitrogen signatures were found to vary with elevation and ant colony size, and between Azteca species groups. Ants in the A. constructor species group appear to be opportunistic foragers at low elevations, but rely more heavily on their host plants at high elevations, whereas ants in the A. alfari species group consume a more consistent diet across their distribution. Further, isotope values indicate that both ant species groups acquire more nitrogen from higher trophic levels at low elevation and when ant colonies are small. Provisioning by the host is a substantial ecological cost to the interaction, and it may vary, even in a highly specialized association. Nonetheless, not all specialized interactions are equivalent; where interaction with one ant species group appears conditional upon the environment, the other is not. Differential host use within the Cecropia Azteca association suggests that the ecological and evolutionary benefits and costs of association may vary among species pairs. PMID- 14564501 TI - The evolution of locomotory behavior in profitable and unprofitable simulated prey. AB - Prey that are unprofitable to attack (for example, those containing noxious chemicals) frequently exhibit slower and more predictable movement than species that lack these defenses. Possible explanations for the phenomenon include a lack of selection pressure on unprofitable prey to avoid predators and active selection on unprofitable prey to advertise their noxiousness. We explicitly tested these and other hypotheses using a novel "artificial world" in which the locomotory characteristics (step size, waiting time, and angular direction) of artificial profitable and unprofitable computer-generated prey were subject to continued selection by humans over a number of generations. Unprofitable prey evolved significantly slower movement behavior than profitable prey when they were readily recognized as unprofitable, and also when they frequently survived predatory attacks. This difference arose primarily as a consequence of more intense selection on profitable prey to avoid capture. When unprofitable prey were very similar (but not identical) in morphological appearance to profitable prey, unprofitable prey evolved particularly slow movement behavior, presumably because when they were slow-moving they could be more readily recognized as being unprofitable. When unprofitable prey were constrained to move slowly, a morphologically identical profitable prey species evolved locomotor mimicry only when it had no more effective means of avoiding predation. Overall, our results provide some of the first empirical support for a number of earlier hypotheses for differences in movement between unprofitable and profitable prey and demonstrate that locomotor mimicry is not an inevitable outcome of selection even in morphologically similar prey. PMID- 14564502 TI - Social rank, feeding and winter weight loss in red deer: any evidence of interference competition? AB - During winter at northern latitudes, large herbivores often exploit patches of concentrated, relatively high quality forage, which may lead to interference competition. The factors affecting success in contests and subsequent dominance rank, such as age and body weight, remain key issues in ungulate behavioural ecology. Maternal effects on offspring body weight are well known, but few studies have investigated if mother's social rank influence offspring rank. Moreover, no study has related dominance rank in ungulates to weight loss during winter. Outcomes of social interactions (n=7,609), feeding time and spatial position in red deer (Cervus elaphus) hinds and calves, and weight loss of calves, were registered from 1981 to 1996 at six winter-feeding sites within the county of Sor-Trondelag in Norway. The level of aggressiveness was higher among calves than among adult hinds, and the factors determining the outcome of contests also differed. The initiator won the majority of interactions (more than 90% in both hinds and calves). Social rank was related to both age and body weight in adult hinds, and related to body weight and mother rank in calves. The relationship between feeding time and rank was non-linear. Feeding time was correlated with rank only among high ranked hinds, while there was no such relationship among low ranked hinds or calves. There was no correlation between winter weight loss and social rank in calves. Our study therefore underlines that, although frequent aggression is observed at artificial feeding sites of northern herbivores, this is not necessarily sufficient to give rise to interference competition. PMID- 14564503 TI - Alpha1beta1-integrin is an essential signal for neurite outgrowth induced by thrombospondin type 1 repeats of SCO-spondin. AB - In the central and peripheral nervous systems a heterogeneous group of proteins constituting the thrombospondin superfamily provides a cue for axonal pathfinding. They either contain or are devoid of the tripeptide RGD, and the sequence(s) and mechanism(s) which trigger in vitro their neurite-promoting activity have remained unclear. In this study, we reconsider the problem of whether sequences present in the thrombospondin type 1 repeats (TSRs), and independent of the well-known RGD-binding site, may activate integrins and account for their neurite-promoting activity. SCO-spondin is a newly identified member of the thrombospondin superfamily, which shows a multidomain organization with a great number of TSR motifs but no RGD sequence. Previous research has implicated oligopeptides derived from SCO-spondin TSRs in in-vitro development of various neuronal cell types. In this study, we investigate whether function blocking antibodies directed against integrin subunits can block these effects in cell line B104, cloned from a neuroblastoma of the rat central nervous system. By two different approaches: flow cytometry revealing short-term effects and cell cultures revealing long-term effects, we show that: (a). activation of cell metabolism, (b). changes in cell size and structure, and (c). neurite-promoting activity induced by TSR oligopeptides are inhibited by function-blocking antibodies to beta1-subunit. Using a panel of function-blocking antibodies directed against various integrin alpha-subunits we show that the alpha1-subunit might be the partner of the beta1-subunit in B104 cells. Thus, we demonstrate that an original sequence within a TSR motif from SCO-spondin promotes neurite outgrowth through an intracellular signal driven by integrins, independently of an RGD-binding site. PMID- 14564504 TI - Continual assembly of desmosomes within stable intercellular contacts of epithelial A-431 cells. AB - Subclones of human carcinoma-derived A-431 cell line stably producing fusion proteins consisting of the enhanced green fluorescent protein and either human desmoglein 2 (Dsg-GFP) or human plakoglobin (GFP-Pg) were used to examine the behavior of desmosomes in living cells. Immunofluorescence microscopy of the fixed cells showed that both fusion proteins, which were expressed in significantly lower levels relative to their endogenous counterparts, were efficiently recruited into desmosomes. Time-lapse confocal imaging of these cells reveals that such GFP-labeled desmosomes (GFP desmosomes) are stable structures which exhibit various dynamic and motile activities. The most notable are independent lateral mobility and fusion. Furthermore, the continual assembly of new nascent desmosomes is observed within stable contacts located at the middle of the epithelial sheet. A new GFP desmosome appears as a closely apposed group of fine patches which after a few minutes aggregate into a single structure. These three dynamic processes resulted in constant changes of desmosome distribution, numbers, and sizes. In addition, fluorescence recovery after photobleaching experiments showed that fine patches of desmosomal proteins may participate in desmosome maintenance. Such a diverse range of dynamic activities of desmosomes apparently produces flexible but tight cell-cell adhesion required for different morphogenetic events in epithelial structures. PMID- 14564505 TI - Analysis of the transcriptional response to Rice Yellow Mottle Virus infection in Oryza sativa indica and japonica cultivars. AB - Several cDNA libraries were constructed using mRNA isolated from roots, panicles, cell suspensions and leaves of non-stressed Oryza sativa indica (IR64) and japonica (Azucena) plants, from wounded leaves, and from leaves of both cultivars inoculated with Rice Yellow Mottle Virus (RYMV). A total of 5549 cleaned expressed sequence tags (ESTs) were generated from these libraries. They were classified into functional categories on the basis of homology, and analyzed for redundancy within each library. The expression profiles represented by each library revealed great differences between indica and japonica backgrounds. EST frequencies during the early stages of RYMV infection indicated that changes in the expression of genes involved in energy metabolism and photosynthesis are differentially accentuated in susceptible and partially resistant cultivars. Mapping of these ESTs revealed that several co-localize with previously described resistance gene analogs and QTLs (quantitative trait loci). PMID- 14564506 TI - Isolation and characterization of a phospholipase C delta isoform from pea that is regulated by light in a tissue specific manner. AB - Phosphoinositide-specific phospholipases C (PLCs) play an important role in many cellular responses and are involved in the production of secondary messengers. We report the cloning and characterization of a cDNA encoding a PLC-delta from Pisum sativum (PsPLC). The amino acid sequence deduced from the cDNA sequence showed 75 80% identity to other plant PLCs and contained the characteristic X, Y and C2 domains. The genomic PLC clone from pea was also characterized and found to contain eight introns. The protein was expressed in Escherichia coli, but the recombinant product did not show any phosphoinositide (PI)- or phosphatidylinositol-4, 5-bisphosphate (PIP2)-specific activity, despite having all known residues required for such activity, and in spite of the fact that its C2 domain was shown to bind calcium. Under similar in vitro assay conditions the recombinant tobacco PLC used as a control showed calcium-dependent PI- and PIP2 specific activity. Though PsPLC did not show enzyme activity in vitro, and may represent an inactive form of PLC, such as those reported in some mammalian systems, analysis of the transcription of PsPLC showed that the gene is expressed in all pea tissues, and is regulated by light in a tissue-specific manner. Roots showed higher expression of PsPLC than shoots. A putative PsPLC promoter region (792 bp) was also cloned and found to contain root-specific and light-responsive cis elements, suggesting that this form of PLC may be involved in important functions in plants. PMID- 14564507 TI - Structural plasmid evolution as a result of coupled recombinations at bom and cer sites. AB - We have studied the recombination of plasmids bearing bom and cer sites. The bom ( basis of mobilization) site is required for conjugative transfer, while the cer ( Col E1 resolution) site is involved in the resolution of plasmid multimers, which increases plasmid stability. We constructed a pair of parent plasmids in such a way as to allow us select clones containing recombinant plasmids directly. Clone selection was based on the McrA sensitivity of recipient host DNA modified by M. Ecl18kI, which is encoded by one of the parent plasmids. The recombinant plasmid contains segments originating from both parental DNAs, which are bounded by bom and cer sites. Its structure is in accordance with our previously proposed model for recombination mediated by bom and cer sequences. The frequency of recombinant plasmid formation coincided with the frequency of recombination at the bom site. We also show that bom-mediated recombination in trans, unlike in cis, is independent of other genetic determinants on the conjugative plasmids. PMID- 14564509 TI - Scanning electron microscopy of Aspidogaster ijimai Kawamura, 1913 and A. conchicola Baer, 1827 (Aspidogastrea, Aspidogastridae) with reference to their fish definitive-host specificity. AB - Two species of aspidogastreans, namely Aspidogaster ijimai and A. conchicola, were studied by scanning electron microscopy. In nine lakes and an old river course, the Tian'ezhou oxbow, investigated in the flood plain of the Yangtze River, A. ijimai was obtained from the common carp ( Cyprinus carpio) in three lakes, and A. conchicola from the black carp Mylopharyngodon piceus in three lakes and the oxbow. In none of the localities, however, were the two species found together. It is suggested that A. ijimai may be considered as a specialist parasite for the common carp, at least in the flood-plain lakes of the Yangtze River. The two parasites were similar in many aspects of their morphology. Their bodies can both be separated into a dorsal part and a ventral disc, with the body surface of the dorsal part elevated by transverse folds, and the disc subdivided into alveoli by transverse and longitudinal septa, although the number of alveoli was different in the two species. The depression on the ventral surface of the neck region was prominent for both species, and their ventral disc was covered densely with non-ciliated bulbous papillae. The position of mouth, osmo regulatory pore and marginal organ was also similar for A. ijimai and A. conchicola. However, microridges in the trough of the folds in the neck region and numerous small pits on the upper part of the septa were found exclusively in A. ijimai, but uniciliated sensory papillae in A. conchicola. PMID- 14564508 TI - Identification and expression analysis of ABC genes in Plasmodium yoelii and P. berghei. AB - The ATP-binding cassette (ABC) proteins are one of the largest evolutionarily conserved families. They are characterized by the presence of highly conserved nucleotide-binding sites (NBS). In the present study, we identified ABC genes in rodent Plasmodia. We queried the Plasmodium yoelii genome with the ABC signature motif and retrieved 15 contigs. Sequences were classified into seven ABC families by BLAST comparison. Conservation of the five signature ABC motifs in the P. yoelii contigs was examined by multi-alignment of the NBS. Expression of the ABC genes was examined during the blood stages of P. yoelii and P. berghei and the hepatocytic stages of P. yoelii. Our results with RT-PCR on total RNA from blood stages demonstrated the expression of 14 ABC genes in P. yoelii and ten in P. berghei. In P. yoelii hepatocytic stages, the expression of four ABC genes was detected. PMID- 14564511 TI - Natural infections of Omphiscola glabra (Lymnaeidae) with Fasciola hepatica in central France. AB - As larval forms of Fasciola hepatica have periodically been detected in Omphiscola glabraafter their collection from watercress beds or from meadows since 1995, field investigations in 37 populations of O. glabra were carried from 1996 to 2002. This was done in order to determine the changes in prevalences and intensities of these natural infections with F. hepatica in relation to the type of snail habitat and the year of snail collection. Snails infected with F. hepatica were found in all samples made in swampy meadows and roadside ditches in all years. In fenced pools and walled gardens, snail infections were only found from 1998 and 1999 onwards, respectively. In the four types of habitats, the prevalences of F. hepatica infections increased slightly over time (0.8-2.1% for snails sampled in swampy meadows, for example) but this increase varied with the habitat. The mean shell heights of infected snails (6.2-7.8 mm) were similar whatever the type of habitat. The numbers of cercariae-containing rediae counted in snails sampled in swampy meadows, roadside ditches, and fenced pools significantly increased over time. Significant numerical variation between these redial burdens was also observed in relation to snail habitat. As the larval development of F. hepatica is facilitated by the presence of another trematode larval form ( Paramphistomum daubneyi), the finding of some naturally infected O. glabra in watering places known to have no contact with domestic or wild large mammals might be explained by the development of P. daubneyi in small mammals such as lagomorphs. However, a progressive adaptation of F. hepatica miracidia to O. glabra over time, which would permit the infection of snails at sizes larger than 2 mm, could not be excluded. PMID- 14564510 TI - Protective immunity against Toxoplasma gondii in mice induced by the SAG2 internal image of anti-idiotype antibody. AB - A monoclonal antibody TGCM5 (isotype G, subclass 1, kappa light chain) against SAG2, a major surface antigen of Toxoplasma gondii tachyzoites, was produced in quantity and its Fab-containing idiotype (Id) was injected into rabbits for production of rabbit anti-Id sera. The rabbit anti-Id IgG (aId-IgG) with a SAG2 internal image was obtained by purification with a protein A column. Significant lymphocyte proliferation induced by tachyzoite lysate antigen was observed in BALB/c mice that received two injections of aId-IgG, but not in non-immunized control mice. Large amounts of gamma interferon were detected in supernatants of splenocyte cultures prepared from mice immunized with aId-IgG. Of the mice immunized with aId-IgG, 75-87.5% survived at least 28 days after a lethal challenge of 1 x 10(3) live tachyzoites, which killed all non-immunized control mice within 10 days. The results showed that the SAG2 internal image presented by aId-IgG indeed elicited a protection against the infection of T. gondii in mice. PMID- 14564512 TI - A model for the transmission of Echinococcus multilocularis in Hokkaido, Japan. AB - A mathematical model for Echinococcus multilocularis transmission would be useful for estimating its prevalence and determining control strategies. We propose a mathematical model which quantitatively describes the transmission of E. multilocularis in Hokkaido, Japan. The model takes into account the influence of the dynamics of both the definitive and the intermediate host populations, which show large scale seasonal variation as they are wild animals. The simulations based on the model clarify the mechanism of seasonal transmission of E. multilocularis quantitatively, notwithstanding a lack of seasonal prevalence data. At present, human alveolar echinococcosis is prevalent throughout the mainland of Hokkaido. The risk of being infected in the human population has been investigated by analyzing the seasonal fluctuation in parasite egg dispersion in the environment. This is necessary for the planning of more suitable preventive measures against E. multilocularis. PMID- 14564513 TI - The current and future role of dexrazoxane as a cardioprotectant in anthracycline treatment: expert panel review. AB - This article summarizes the views of an expert meeting of cardiologists and oncologists on the use of dexrazoxane in anthracycline-based chemotherapy. Anthracycline-induced cardiotoxicity remains a major concern and new trends in treatment (e.g., combination of an anthracycline with other agents) will ensure that it remains a problem. Dexrazoxane reduces this cardiotoxicity in adults and children with a range of tumor types. Further research may help to identify those patients who are at particular risk of cardiotoxicity and who would benefit the most from dexrazoxane. There are also numerous possibilities for dexrazoxane in other clinical situations, which must be addressed in future trials. PMID- 14564514 TI - Increased expression of CEA and MHC class I in colorectal cancer cell lines exposed to chemotherapy drugs. AB - PURPOSE: Cancer-specific immunotherapy holds great promise as an emerging treatment for advanced colorectal cancer and may be combined with standard chemotherapy to provide a synergistic inhibitory action against tumor cells. To examine the interrelationship between the immune system and chemotherapy, we studied the induction of both CEA, a tumor-associated antigen, and MHC class I, a major component of the antigen presenting system, in response to a number of chemotherapeutic agents. METHODS: The effect of a selection of standard chemotherapeutics on MHC class I and CEA expression in human colorectal cancer cell lines was determined by flow cytometry and semi-quantitative RT-PCR. In addition, studies using mice bearing tumors derived from an injected murine colon cancer cell line were performed to determine if alteration in MHC class I expression occurs in vivo following continuous infusion of chemotherapeutic agents into the peritoneal cavity, as well as to facilitate correlations between expression of this factor and therapeutic effectiveness. RESULTS: All anti-cancer drugs examined, when given at IC50 values, induced expression of MHC class I protein in the human colon cancer cell line, COLO201. However, expression of CEA mRNA was only induced upon exposure to 5-FU, in contrast to obscure induction following CDDP and SN-38 treatment. Combined treatment with 5-FU and CDDP gave additional effect on CEA expression in COLO201 cells. Regarding the in vivo studies in mice, the size of the murine colon cancer cell-derived tumors was reduced only in response to treatment with CDDP, which also mediated the highest induction of MHC class I expression. CONCLUSION: These results suggest that chemotherapeutic agents trigger the immune system and cancer-specific immunotherapy may be effective when used in combination with systemic chemotherapy. PMID- 14564515 TI - EphA2 up-regulation induced by deoxycholic acid in human colon carcinoma cells, an involvement of extracellular signal-regulated kinase and p53-independence. AB - PURPOSE: The EphA2 receptor protein tyrosine kinase gene has been shown to be over-expressed or functionally altered in a number of human tumors, including colon cancer, but little is known about the regulation of this new oncoprotein. In order to explore the mechanism of EphA2 up-regulation in cancer cells, we examined the change of expression of EphA2 gene induced by deoxycholic acid (DCA) and elucidated its possible pathways in human colon cancer cells. METHODS: Western blot and RT-PCR were used to assess the protein expression and messenger RNA in several colon cancer cell lines, which harbor various p53 status. The inhibition study to interfere the MAPK pathway was performed by using various chemicals and by transfecting dominant negative mutant plasmids. RESULTS: Up regulation of EphA2 induced by DCA was observed in a dose- and time-dependent fashion both in mRNA and protein levels. This regulation is constant regardless of p53 status including wild, mutant or knocked out in the colon cell lines used. This induction was in part blocked by either erk1/2 inhibitors or dominant negative mutants erk1/2 plasmids. CONCLUSIONS: These results suggest that DCA induced up-regulation of EphA2 in colon cancer cells is due to activation of erk1/2 cascade, and is p53-independent. Taken together with the roles of EphA2 and DCA in tumorigenesis, which have been independently reported, our observation will provide a new mechanistic basis of DCA commitment in carcinogenesis. PMID- 14564516 TI - Initial experience with a novel desensitization strategy for carboplatin associated hypersensitivity reactions: carboplatin-hypersensitivity reactions. AB - PURPOSE: Carboplatin hypersensitivity is an increasingly recognized toxicity in individuals receiving >6 cumulative courses of this important antineoplastic agent. We wished to determine if a novel multi-pronged approach to re-treating patients with a high risk for this potentially serious side effect could permit the safe delivery of this class of cytotoxic drugs. METHODS: Five patients with gynecologic malignancies who had either experienced a documented carboplatin hypersensitivity reaction ( n =4) or had a "positive" carboplatin skin test ( n =1), received a multi-drug oral regimen administered over several days which was designed to block known mediators of anaphylaxis. Four of these individuals subsequently underwent treatment with either cisplatin or carboplatin employing a "dose escalation" desensitization schema. RESULTS: Four patients underwent successful treatment with either cisplatin or carboplatin (3, 4, 5, 6+ total additional courses) without any further evidence of hypersensitivity. CONCLUSION: In this preliminary report of a limited patient population, we have demonstrated the ability to safely deliver a platinum agent to individuals with either documented carboplatin hypersensitivity, or a high risk for this potentially serious toxicity of carboplatin. Further exploration of this novel management strategy in a larger group of patients is indicated. PMID- 14564517 TI - Malignant hypertension and cerebral haemorrhage in Seckel syndrome. AB - Seckel syndrome is an autosomal recessive condition with severe short stature and facial and neurological anomalies. Intracranial haemorrhage, due to rupture of a cerebral aneurysm, is a very rare complication of this syndrome. Malignant hypertension may play an important role in the aetiology of the aneurysm and early detection is essential in order to prevent organ damage. CONCLUSION: we report a new case of Seckel syndrome associated with malignant hypertension and cerebral haemorrhage. PMID- 14564518 TI - Water retention in the acute phase of Kawasaki disease: relationship between oedema and the development of coronary arterial lesions. AB - Despite intravenous immunoglobulin therapy, a certain percentage of patients with Kawasaki disease (KD) still develop coronary arterial lesions (CAL). In an effort to find new combined therapies to reduce the incidence of CAL, we focused on the oedema which can be an important sign of the increased vascular permeability in KD. A total of 127 patients with KD were included in the retrospective study. Serial weekly changes in serum sodium and albumin levels from the 1st to the 4th week of illness were examined. In addition, the maximum rate of increase in body weight from admission to the 14th day of illness was evaluated. Serum sodium levels (mEq/l) in only the 2nd week of illness were significantly lower in patients with CAL than in those without CAL (mean +/- SD, 135.5+/-4.5 versus 138.0+/-2.4, P<0.05). Serum albumin levels in all 4 weeks were significantly lower in patients with CAL than in those without CAL ( P<0.001). The maximum rate (%) of increase in body weight from admission to the 14th day of illness was significantly higher in patients with CAL than in those without CAL (ranges and median values, 0-12.3 (7.0) versus 0-10.3 (3.2), P<0.001). CONCLUSION: these results suggest that water retention in the acute phase of Kawasaki disease may be a risk factor for CAL, and water intake of both infusion and oral intake should be kept to a minimum in order to avoid progressive oedema. PMID- 14564520 TI - ZWILLE buffers meristem stability in Arabidopsis thaliana. AB - The shoot apical meristem of higher plants consists of a population of stem cells at the tip of the plant body that continuously gives rise to organs such as leaves and flowers. Cells that leave the meristem differentiate and must be replaced to maintain the integrity of the meristem. The balance between differentiation and maintenance is governed both by the environment and the developmental status of the plant. In order to respond to these different stimuli, the meristem has to be plastic thus ensuring the stereotypic shape of the plant body. Meristem plasticity requires the ZWILLE (ZLL) gene. In zll mutant embryos, the apical cells are misspecified causing a variability of the meristem's size and function. Using specific antibodies against ZLL, we show that the zll phenotype is due to the complete absence of the ZLL protein. In immunohistochemical experiments we confirm the observation that ZLL is solely localized in vascular tissue. For a better understanding of the role of ZLL in meristem stability, we analysed the genetic interactions of ZLL with WUSCHEL (WUS) and the CLAVATA1, 2 and 3 (CLV) genes that are involved in size regulation of the meristem. In a zll loss-of-function background wus has a negative effect whereas clv mutations have a positive effect on meristem size. We propose that ZLL buffers meristem stability non-cell-autonomously by ensuring the critical number of apical cells required for proper meristem function. PMID- 14564519 TI - Expression of ICAM-1 in colon epithelial cells: an ultrastructural study performed on in vivo and in vitro models. AB - BACKGROUND: Studies have suggested that in ulcerative colitis (UC), intercellular adhesion molecule-1 (ICAM-1) is involved in migration of leukocytes toward the colonic epithelium. A suitable in vitro model of chronic colonic inflammation does not exist, and the role of the epithelium is based on monolayers of cancer cells. Conflicting results exist on epithelial ICAM-1 expression, and the aim of this study was to compare the expression in various models of colonic epithelium. MATERIALS AND METHODS: Colonic biopsies from four UC patients and four controls were examined by cryoimmuno-electron microscopy using ICAM-1-antibodies. In four other controls, the epithelium was isolated from colonic biopsies, embedded in collagen, and evaluated similarly. Isolated crypts and cultured cancer cells were stimulated with interferon-gamma (IFN-gamma) or tumor necrosis factor-alpha (TNF alpha). RESULTS: ICAM-1 was not expressed in the biopsies. In contrast, HT29 cells and the collagen-embedded crypts expressed ICAM-1 on the apical membranes proximal to the junctional complexes when stimulated with IFN-gamma or TNF-alpha in a dose-related manner. CONCLUSIONS: ICAM-1 is not expressed on colonic epithelium in vivo. However, both colonocytes and HT29 cells were capable of expressing ICAM-1 on their apical membranes in response to supraphysiologic cytokine concentrations. These observations question the justification of extrapolating observations from colon cancer cell lines to in vivo inflammatory conditions. PMID- 14564521 TI - Cyanobacterial H(2) production -- a comparative analysis. AB - Several unicellular and filamentous, nitrogen-fixing and non-nitrogen-fixing cyanobacterial strains have been investigated on the molecular and the physiological level in order to find the most efficient organisms for photobiological hydrogen production. These strains were screened for the presence or absence of hup and hox genes, and it was shown that they have different sets of genes involved in H(2) evolution. The uptake hydrogenase was identified in all N(2)-fixing cyanobacteria, and some of these strains also contained the bidirectional hydrogenase, whereas the non-nitrogen fixing strains only possessed the bidirectional enzyme. In N(2)-fixing strains, hydrogen was mainly produced by the nitrogenase as a by-product during the reduction of atmospheric nitrogen to ammonia. Therefore, hydrogen production was investigated both under non-nitrogen fixing conditions and under nitrogen limitation. It was shown that the hydrogen uptake activity is linked to the nitrogenase activity, whereas the hydrogen evolution activity of the bidirectional hydrogenase is not dependent or even related to diazotrophic growth conditions. With regard to large-scale hydrogen evolution by N(2)-fixing cyanobacteria, hydrogen uptake-deficient mutants have to be used because of their inability to re-oxidize the hydrogen produced by the nitrogenase. On the other hand, fermentative H(2) production by the bidirectional hydrogenase should also be taken into account in further investigations of biological hydrogen production. PMID- 14564523 TI - Constitutive nitric oxide synthases in rat pancreatic islets: direct imaging of glucose-induced nitric oxide production in beta-cells. AB - The production of nitric oxide (NO) by constitutive nitric oxide synthase (NOS) was investigated in isolated rat pancreatic islets and dispersed beta-cells. Double-immunocytochemical analyses with a confocal microscope demonstrated the presence of NOS1 in alpha-, beta-, delta-, and PP-cells and that of NOS3 in beta , delta-, and PP-cells, but not alpha-cells, in the isolated rat islets. Image analyses with the NO-reactive fluorescence dye DAF-2 clearly showed that an elevation in glucose concentrations from 0 to 11.1 mM increased intracellular NO in most cells of the isolated islets. The glucose-induced elevation of intracellular NO in the islet cells was abolished in the presence of the Ca2+ channel blocker nicardipine and after treatment with the NOS inhibitor NG-nitro-L arginine. Similarly, the ATP-sensitive K+ channel blocker tolbutamide, which elevates intracellular Ca2+ concentrations, increased DAF-2 fluorescence in most cells of the isolated islets. In isolated beta-cells, 11.1 mM glucose increased DAF-2 fluorescence, which was suppressed by NG-nitro-L-arginine and by reducing the glucose concentration to 0 mM. DAF-2 fluorescence in beta-cells was also increased by 50 mM K+, which was suppressed by NG-nitro-L-arginine. These results suggest that NOS1 and NOS3 are present in rat pancreatic beta-cells, and that glucose produces NO by Ca(2+)-dependent activation of the constitutive NOS isoforms. PMID- 14564522 TI - Characterization of a protein of the plastid inner envelope having homology to animal inorganic phosphate, chloride and organic-anion transporters. AB - A protein from Arabidopsis thaliana (L.) Heynh. showing homology to animal proteins of the NaPi-1 family, involved in the transport of inorganic phosphate, chloride, glutamate and sialic acid, has been characterized. This protein, named ANTR2 (for anion transporters) was shown by chloroplast subfractionation to be localized to the plastid inner envelope in both A. thaliana and Spinacia oleracea (L.). Immunolocalization revealed that ANTR2 was expressed in the leaf mesophyll cells as well as in the developing embryo at the upturned-U stage. Five additional homologues of ANTR2 are found in the Arabidopsis genome, of which one was shown by green fluorescent protein (GFP) fusion to be also located in the chloroplast. All ANTR proteins share homology to the animal NaPi-1 family, as well as to other organic-anion transporters that are members of the Anion:Cation Symporter (ACS) family, and share the main features of transporters from this family, including the presence of 12 putative transmembrane domains and of a 7 amino acid motif in the fourth putative transmembrane domain. ANTR2 thus represent a novel protein of the plastid inner envelope that is likely to be involved in anion transport. PMID- 14564524 TI - Neuromuscular response to sustained low-level muscle activation: within- and between-synergist substitution in the triceps surae muscles. AB - The intent of this study was to investigate physiological recruitment strategies employed by the triceps surae (TS) muscles during sustained low-level activation. Surface-recorded electromyographic (EMG) signal data were recorded from the medial gastrocnemius (MG), lateral gastrocnemius (LG), and medial soleus (MS) muscles while eight subjects sustained static plantarflexion contractions at 10% of their maximum voluntary contraction (MVC) for 1 h. Fine-wire EMG activity was simultaneously recorded from electrodes located at three sites within the MG muscle. Correlation coefficients were computed among root mean square (RMS) amplitude values recorded from MG, LG, and MS to investigate between-synergist substitution, as well as between the three wire channels to investigate local substitution within the MG motor unit pool. Over the 1-h test, EMG amplitude in LG and MS increased linearly, and there was a moderately strong ( R(2)=0.662, P<0.023) positive correlation in the detrended activity between LG and MS, suggesting that these muscles generally acted together. When the data were divided into 5-min blocks, regression analysis on the partial correlation data revealed that MS and LG were correlated over the duration of the contraction ( P<0.001) suggesting co-activation synergism, whereas MG and LG and MS and MG demonstrated a tendency toward trade-off synergism. In five of eight individuals at least one wire channel pair within the MG was negatively correlated, suggesting that there was some form of substitution between motor units or motor unit pools. The other three individuals maintained correlated activity between all three pairs throughout the 1-h contraction. PMID- 14564525 TI - Gait transition cost in humans. AB - The energetics of locomotion depend largely on speed, gait and body size. Gait selection for a given speed appears partly, but perhaps not wholly, related to metabolic cost. One cost normally omitted from considerations of locomotion efficiency is the metabolic cost of the transition between gaits. We present the first direct assessment of the metabolic cost for the walk-run/run-walk transition in humans. The average increase in metabolic cost for a step involving a transition is 1.75 times that of a mean non-transition step at a speed where metabolic power requirements are identical for walking and running. Despite this substantial increase in cost for the transition step, the metabolic cost of gait transition is unlikely to have a strong bearing on the process of gait selection as the cost of using a metabolically inappropriate gait, even for only a few steps, will dominate. PMID- 14564526 TI - Test-retest errors and the apparent heterogeneity of training response. AB - Published reports have shown large apparent inter-individual differences of gains in maximal oxygen intake (VO(2max)) in response to a standard 20-week programme of aerobic conditioning that progressed to 75% of the individual's initial VO(2max). The observed gains of VO(2max) ranged from 0 to 1,000 ml min(-1), with a coefficient of variation (CV) of 8.4%. The present analysis evaluates the potential contribution of test-retest errors to these apparent large inter individual differences in training response. The 2-day test-retest CV for VO(2max) readings in 742 healthy adults was initially 5.0%, dropping to 4.1% after training. Published training responses were estimated from the mean of paired measurements obtained before and after training if readings agreed by <5%, but from the highest of paired values if these differed by >5%. Taking account of the relative proportions of single and paired observations, the weighted VO(2max) data for the entire sample had an effective 2-day CV of 4.3% before and 3.4% after training. Assumption 1: if the 20-week test-retest error remained similar to the 2-day figure, measurement error would contribute a CV of 5.5% to apparent training responses, or (for the stated initial mean VO(2max) of 2,409 ml min(-1)) an SD of 132 ml min(-1). Assumption 2: if the 20-week CV was similar to that in other long-term studies (~5%), measurement error would contribute a CV of 6.1%, or a SD of 146 ml min(-1). The published data show a total SD of 202 ml min(-1) for apparent inter-individual differences in training response, with age, gender, race and baseline VO(2max) accounting for only 11% of this variance. After estimating the likely effect of test-retest measurement errors, the SD due to inter-individual differences would decrease to 138 ml min VO(2max) (assumption 1) or 123 ml min(-1) (assumption 2). We conclude that when estimating the extent of inter-individual differences in training response, allowance must be made not only for the minor effects of recognized covariates (age, gender, race and initial fitness), but also for the larger influence of test-retest measurement errors. Nevertheless, substantial inter-individual differences persist after making such adjustments. The most likely explanation of these differences is a familial aggregation of training responses. PMID- 14564527 TI - Urinary hydroxy-metabolites of naphthalene, phenanthrene and pyrene as markers of exposure to diesel exhaust. AB - OBJECTIVE: The objective of this study was to assess the exposure of bus-garage and waste-collection workers to polycyclic aromatic hydrocarbons (PAHs) derived from diesel exhaust by the measurement of levels of seven urinary PAH metabolites: 2-naphthol, 1-hydroxyphenanthrene, 2-hydroxyphenanthrene, 3 hydroxyphenanthrene, 1+9-hydroxyphenanthrene, 4-hydroxyphenanthrene and 1 hydroxypyrene. SUBJECTS AND METHODS: One urine sample from each of 46 control persons, and one pre-shift and two post-shift spot urine samples from 32 exposed workers were obtained in winter and in summer. The metabolites were analysed after enzymatic hydrolysis by high performance liquid chromatography (HPLC) with fluorescence detection. RESULTS: The sum of seven PAH metabolites (mean 3.94 +/- 3.40 and 5.60 +/- 6.37 micromol/mol creatinine in winter and summer, respectively) was higher [P=0.01, degrees of freedom (df) =61.2 and P=0.01, df=67.6 in winter and summer, respectively] in the exposed group than in the control group (mean 3.18 +/- 3.99 and 3.03 +/- 2.01 micromol/mol creatinine in winter and summer, respectively). The mean concentrations of 2-naphthol among exposed and controls ranged between 3.34 and 4.85 micromol/mol creatinine and 2.51 and 2.58 micromol/mol creatinine, respectively (P<0.01 in winter, P<0.03 in summer). The mean level of the hydroxyphenanthrenes in the samples of exposed workers was between 0.40 and 0.70 micromol/mol creatinine and in the control samples 0.40-0.60 micromol/mol creatinine. The concentration of 1-hydroxypyrene was higher among exposed workers in both pre-shift and post-shift samples (mean 0.10-0.15 micromol/mol creatinine) than in control group (mean 0.05-0.06 micromol/mol creatinine) in winter (P=0.002, df=78) and in summer (P<0.001, df=68). CONCLUSIONS: The urinary hydroxy-metabolites of naphthalene, phenanthrene and pyrene showed low exposure to diesel-derived PAHs; however, it was higher in exposed workers than in control group. Urinary PAH monohydroxy-metabolites measured in this study did not correlate with the PAHs in the air samples, reported earlier, in 2002 and 2003. PMID- 14564528 TI - The thyroid C cells of ovariectomized rats treated with estradiol. AB - The structure and function of thyroid C cells were studied in ovariectomized (Ovx) adult female rats without and after chronic treatment with estradiol dipropionate (EDP). A peroxidase-antiperoxidase method was applied for localization of calcitonin (CT) in the C cells. Morphometric changes in their volume, nuclei, and relative volume density were evaluated in comparison with sham-operated control rats using a stereological method. The number of C cells was calculated. CT content in the sera was determined by radioimmunoassay. Ovariectomy (Ovx) led to a 21% increase in body weight ( P<0.005), while treatment of Ovx rats with EDP decreased body weight by 25% ( P<0.01). The immunoreactivity for CT in C cells of the Ovx rats was markedly increased. Significant decreases in the volume of C cells (by 13%; P<0.05) and serum CT (by 45%) were recorded, while the C cell number increased by 59% ( P<0.05) in relation to the corresponding controls. The treatment of Ovx rats with EDP caused conspicuous degranulation of the C cells. The cellular volume was increased by 11% and serum CT by 36% in comparison with Ovx animals. At the same time a decrease in C cell number by 29% ( P<0.05) was evident. It may be concluded that estradiol deficiency after Ovx reduced the synthesis and release of CT, while chronic treatment of these animals with EDP had a positive effect on the secretory activity of thyroid C cells. PMID- 14564530 TI - Comparison of long-term fluctuations: laser scanning tomography versus automated perimetry. AB - BACKGROUND: The aim of our study was to investigate the long-term variability of laser scanning tomography and to compare it with the long-term variability of automated perimetry. METHOD: We retrospectively reviewed the charts of 199 patients (394 eyes) who underwent both automated perimetry (Octopus) and laser scanning tomography (Heidelberg retina tomograph, HRT) in our hospital since 1994. All patients either had ocular hypertension (OHT) or early stage glaucoma. The mean follow-up time was 5.4 years; the average number of examinations was 7.7. The variability was described by the variance ratio VR. RESULTS: The HRT parameters showed very similar long-term fluctuations as the Octopus parameters. In detail: VR of HRT parameters ranged from 0.04 (cup area, cup volume, mean cup depth) to 0.35 (contour height variation), whereas VR of Octopus parameters ranged from 0.12 (mean sensitivity) to 0.15 (loss variance). CONCLUSIONS: The long-term variability of HRT parameters is in the same range as the long-term variability of visual field parameters. Since it is now widely accepted that visual field changes over time should be reproduced at least once or twice before clinical consequences can be drawn, the same should be postulated for HRT changes over time. PMID- 14564529 TI - Localisation of P2Y1 and P2Y4 receptors in dorsal root, nodose and trigeminal ganglia of the rat. AB - The presence and distribution of P2Y (nucleotide) receptor subtypes in rat sensory neurons has been investigated. RT-PCR showed that P2Y(1), P2Y(2), P2Y(4) and P2Y(6) receptor mRNA is expressed in sensory ganglia [dorsal root ganglion (DRG), nodose ganglion (NG) and trigeminal ganglion (TG)]. The regional and cellular distribution of P2Y(1) and P2Y(4) receptor proteins in these ganglia was investigated using immunohistochemistry. P2Y(1) polyclonal antibodies stained over 80% of the sensory neurons, particularly the small-diameter (neurofilament negative) neurons. The P2Y(4) receptor antibody stained more medium- and large- (neurofilament-positive) diameter neurons than small-diameter neurons. P2Y(1) and P2Y(4) receptor immunoreactivity (P2Y(1)-IR and P2Y(4)-IR) was often coexpressed with P2X(3) receptor immunoreactivity (P2X(3)-IR) in subpopulations of neurons. Double immunohistochemistry showed that 73-84% of P2X(3) receptor-positive neurons also stained for the P2Y(1) receptor in DRG, TG and NG while only 25-35% also stained for the P2Y(4) receptor. Subpopulations of P2Y(1)-IR neurons were coexpressed with NF200, CGRP and IB(4); most P2Y(4)-IR neurons were coexpressed with NF200, while only a few neurons were coexpressed with CGRP (10-20%) or with IB(4) (1-2%). The results suggest that P2Y as well as P2X receptor subtypes contribute to purinergic signalling in sensory ganglia. PMID- 14564532 TI - Association between HLA-A2 in Japanese psoriasis arthritis and susceptibility to uveitis. PMID- 14564531 TI - NADPH diaphorase activity in the rat retina during the early stages of experimental diabetes. AB - BACKGROUND: Nitric oxide (NO) plays an important physiological role in inter cellular communication, but when produced in excess it can become toxic. Our goal was to evaluate possible involvement of NO in the development of retinopathy in diabetic rats. METHODS: Diabetes was induced in male albino rats by intravenous injection of streptozotocin. Some of the normal and diabetic rats were raised with added L-arginine to increase in vivo NO synthesis, or with added L-NAME to inhibit the rate of in vivo NO synthesis. NADPH diaphorase histochemistry was conducted on retinal whole mounts and transverse sections at different time intervals after induction of diabetes. The electroretinogram (ERG) was recorded to assess retinal function. RESULTS: After 6 weeks of diabetes, NADPH diaphorase amacrine cells in the diabetic retinas appeared abnormal in their morphology and the degree of staining was decreased in their processes. In contrast, NADPH diaphorase activity was augmented in Muller cells. Supplementing the rats' diet with L-arginine for 10 weeks slightly reduced NADPH diaphorase activity in amacrine cell in normal rats but had no effect on the diabetic rats. Adding L NAME for 10 weeks did not alter NADPH diaphorase histochemistry in either normal or diabetic rats. The ERG responses were reduced by L-arginine supplementation in normal and diabetic rats, and were unaffected by adding L-NAME to the drinking water. CONCLUSIONS: Our findings are consistent with the hypothesis that high glucose levels are deleterious to the rat retina and that excessive synthesis of NO may contribute to the development of diabetic retinopathy. PMID- 14564533 TI - In vitro culture of the tropical sponge Axinella corrugata (Demospongiae): effect of food cell concentration on growth, clearance rate, and biosynthesis of stevensine. AB - In vitro culture is one possible method for supplying sponge metabolites for pharmaceutical applications, but appropriate feeding regimens that maximize both growth and metabolite biosynthesis are largely unknown. According to the natural concentration (NC) of cells 1 to 50 micro m in size that are available to wild Axinella corrugata, we fed explants a multispecific diet of bacteria, microalgae, and yeast at 4 different concentrations: 1NC, 3NC, 5NC, and 5+1NC (the last consisted of 5 NC of bacteria and 1 NC of microalgae and yeast). Explants fed a 3NC diet had the best culture response, growing on average from 8.5 g to 10.3 g in 8 weeks, and showing a 110% increase in concentration (milligrams per gram of dry weight) of the antitumor compound stevensine. Stevensine production in 3NC explants, representing the total milligrams of metabolite per explant, increased by 157% over the study. Explants fed at 1NC had relatively stable weights, indicating that the diet met metabolic costs only. Explants fed at the two highest concentrations lost weight after 4 weeks, possibly because long-term high cell concentration blocked their aquiferous system, reducing their ability to feed efficiently. Stevensine production in explants fed the 1NC, 5NC, or 5+1NC diets were similar, and varied little from the initial amount. A separate experiment showed that the clearance rate for A. corrugata is similar between the examined food types and cell concentrations over 5 hours, averaging 766 ml h(-1) g DW(-1).Overall, this study demonstrates that relatively small changes in food abundance can greatly affect both sponge growth and metabolite biosynthesis. The good growth and increased production of the target metabolite stevensine for A. corrugata explants fed a 3NC diet suggests that in vitro culture is a viable method of supplying some sponge metabolites. PMID- 14564535 TI - Gynogenesis assessment using microsatellite genetic markers in turbot ( Scophthalmus maximus). AB - Gynogenesis was assessed by different methods in 2 families of gynogenetic offspring in turbot ( Scophthalmus maximus). Karyotype analysis in embryos and larvae demonstrated high accuracy in estimation of ploidy level, but performance was uneven given the low quality and number of plates obtained. The use of silver staining to estimate the number of nucleoli per nucleus resulted in a straightforward and easy method to evaluate the ploidy of the samples studied. However, the existence of a nucleolus organizer region polymorphism in turbot determined a small error in ploidy estimation, important when checking ploidy in specific individuals. The use of a set of 11 highly variable microsatellite loci proved to be a powerful method to confirm the exclusive maternal inheritance to gynogenetic offspring in turbot, with probabilities of detection of putative paternal genetic contribution above 99.99%. PMID- 14564534 TI - AFLP-based genetic linkage maps of the pacific oyster Crassostrea gigas Thunberg. AB - Amplified fragment length polymorphisms (AFLPs) were used for genome mapping in the Pacific oyster Crassostrea gigas Thunberg. Seventeen selected primer combinations produced 1106 peaks, of which 384 (34.7%) were polymorphic in a backcross family. Among the polymorphic markers, 349 were segregating through either the female or the male parent. Chi -square analysis indicated that 255 (73.1%) of the markers segregated in a Mendelian ratio, and 94 (26.9%) showed significant ( P < 0.05) segregation distortion. Separate genetic linkage maps were constructed for the female and male parents. The female framework map consisted of 119 markers in 11 linkage groups, spanning 1030.7 cM, with an average interval of 9.5 cM per marker. The male map contained 96 markers in 10 linkage groups, covering 758.4 cM, with 8.8 cM per marker. The estimated genome length of the Pacific oyster was 1258 cM for the female and 933 cM for the male, and the observed coverage was 82.0% for the female map and 81.3% for the male map. Most distorted markers were deficient for homozygotes and closely linked to each other on the genetic map, suggesting the presence of major recessive deleterious genes in the Pacific oyster. PMID- 14564536 TI - Submersed culture production of extracellular wax esters by the marine bacterium Fundibacter jadensis. AB - Fundibacter jadensis strain T9, a marine gram-negative bacterium, was isolated from the intertidal sediment of the German North Sea coast by our group. The cells were able to produce considerable amounts of extracellular wax esters when cultivated with n-alkanes (hexadecane or tetradecane) as a carbon source. The dependence of wax ester production and the composition of the purified wax on different culture conditions (C:N:P ratio and dissolved oxygen tension) were tested. Our results show that wax ester production was not directly growth linked. The C:N:P ratio had no significant influence on the yield of alkane-free purified wax. The dissolved oxygen tension affected the produced amount of the alkane-free purified wax and the composition of the purified wax; when lower than 2% it decreased the yield of purified wax and led to an altered wax ester composition. Tetradecane as a carbon source enhanced the spectrum of the wax ester composition. PMID- 14564537 TI - Multiple paternity in Norway lobster (Nephrops norvegicus L.) assessed with microsatellite markers. AB - We investigated genetic diversity and the mating system of the Norway lobster (Nephrops norvegicus) in a wild population off the Portuguese coast. Approximately 100 individuals were screened for 2 microsatellite loci. For 11 ovigerous lobsters both the female and a sample of her offspring (24 eggs) were genotyped. High genetic diversity was observed for the 2 markers in the population. Paternity within broods was analyzed by comparing multilocus genotypes of each egg with the corresponding mother, and the male parent contribution was then deduced. Multiple paternity was observed in 6 of the 11 broods studied. In those cases, 2 to 3 male parents were likely to have contributed to the fertilization of the eggs. When multiple paternity was involved, the comparative reproductive success of the male parents was quite even. This is the first report of multiple paternity in the Norway lobster. Comparisons with other taxa are presented, and consequences of multiple paternity are discussed. PMID- 14564538 TI - Virus isolation for diagnosing dengue virus infections in returning travelers. AB - Dengue fever is recognized as one of the most frequent imported acute febrile illnesses affecting European tourists returning from the tropics. In order to assess the value of virus isolation for the diagnosis of dengue fever, 70 cases of dengue fever confirmed in German travelers during the period 1993-2001 were analyzed retrospectively. In 26 patients who had developed acute febrile illness within 2 weeks following their return from a trip to a dengue-endemic area, 9 of 13 attempts to isolate the virus were successful in sera drawn 1-5 days and 2 of 13 sera drawn 6-10 days after the onset of illness. DEN-1 was the most frequent serotype isolated. If performed early, virus isolation is a reliable tool for detecting dengue virus in returning travelers. PMID- 14564540 TI - Human endogenous retroviruses with transcriptional potential in the brain. AB - Genetic studies of neuropsychiatric disorders have often produced conflicting results, which might partly result from the involvement of epigenetic modifications. We intended to explore the possible implication of DNA methylation and human endogenous retroviruses (HERVs) in neuropsychiatric disorders. In the present study, we identified two HERV loci that are expected to retain the transcriptional activity in the brain. One was located on chromosome 1q21-q22 and the other on 22q12. Interestingly, these regions were overlapped with or included in those of schizophrenia-susceptible loci, SCZD9 and SCZD4, respectively. Particularly, the HERV on 22q12 was located in the opposite direction 4 kb downstream of the Synapsin III gene. These HERV loci could afford clear targets for methylation and expression analyses in postmortem brains of patients with psychiatric disorders such as schizophrenia. In addition, we confirmed our previous finding that only a few of particular HERV-K loci were activated among a number of highly homologous loci in teratocarcinoma cell lines. These activated loci included ones common to all teratocarcinoma cell lines analyzed and depending on their male or female origin. PMID- 14564539 TI - Voriconazole salvage treatment of invasive candidiasis. AB - Data on the salvage treatment of invasive candidiasis with voriconazole in 52 patients intolerant of other antifungal agents or with infection refractory to other antifungal agents were analyzed. Patients had received a mean of two previous antifungal agents (range, 1-4 agents), and 83% had received an azole. Manifestations of invasive candidiasis included candidemia (37%), disseminated disease (25%), and infection of other sites (38%). The median duration of voriconazole therapy was 60 days (range, 1-314 days). The overall rate of response was 56% (95%CI, 41-70), with the following response rates observed for individual Candida species: Candida albicans, 44% (20-70); Candida glabrata, 38% (14-68); Candida krusei, 70% (35-93); Candida tropicalis, 67% (30-93); and other Candida spp., 100% (40-100). The response rate in patients who had failed previous azole therapy was 58% (42-73). Common adverse events (~20%) included nausea and emesis, abnormal liver enzymes, and visual disturbances. Serious adverse events occurred in four patients, and nine patients died. Voriconazole has promise as a salvage agent for the treatment of invasive candidiasis, even in the settings of previous azole therapy and infection due to Candida krusei. PMID- 14564542 TI - Hodgkin-like posttransplant lymphoproliferative disorder in children: does it differ from posttransplant Hodgkin lymphoma? AB - Epstein-Barr virus (EBV)-mediated lymphoid proliferations occur in patients who are immunocompromised and are reported following bone marrow or solid organ transplantation. Most post-transplant lymphoproliferative disorders (PTLD) are polymorphic in appearance; some are monomorphic and can resemble conventional malignant lymphomas. PTLD that resembles Hodgkin lymphoma has been reported infrequently. We herein report seven cases of PTLD that have large numbers of Reed-Sternberg-like (RS-like) cells and highlight differences in the phenotype of these cases that may distinguish Hodgkin-like PTLD (HL-PTLD) from true Hodgkin lymphoma/disease (HD). All patients were in the second decade of life and were 8 months to 13 years following transplant. HL-PTLD involves lymph nodes that contain a mixed population of small to intermediate-sized lymphocytes with large mononuclear and occasionally binucleate RS-like cells. The large cells of HL-PTLD are pleomorphic B cells that react strongly for CD20 and/or CD79a, express CD30, but are usually negative for CD15 and have few mitoses. They are positive for EBV early RNA (EBER) using an EBER-1 probe, as are some of the background small lymphocytes. A single case of true Hodgkin lymphoma has highly atypical RS-like cells that contain numerous mitoses, does not have CD20 or CD79a reactivity, has CD15 and CD30 staining, and the EBER-1 probe is confined to the large cells only. All patients were managed by withdrawal of immunosuppression and variably treated with either antiviral or anti-CD20 monoclonal antibody, or with chemotherapy. A unique instance of evolution from a HL-PTLD to true HD is also illustrated. In conclusion, HL-PTLD and HD appear to be two related but immunophenotypically and biologically distinct forms of lymphoproliferation in post-transplant patients and may require different protocols for their management. PMID- 14564550 TI - Operative repair of popliteal aneurysms: effect of factors related to the bypass procedure on outcome. AB - The objective of this study was to compare patency rates following the repair of popliteal aneurysms according to the site of inflow, material of bypass graft and quality of distal runoff. Seventy bypasses were performed over an 11-year period. Autogenous saphenous vein was used in 53 procedures (75.7%) and prosthetic material was used in 17 (24.3%). Early mortality was 2.8%. Early primary and secondary patency rates were 95.7% and 97.1%, respectively. Autogenous vein showed better 10-year patency than prosthetic material (86% vs. 57%; p = 0.02). No significant differences in patency were observed according to the inflow site (87.8% groin vs. 74.7% supragenicular). Bypasses that originated in the groin showed improved patency when a saphenous vein was used (84.8% vs. 43.7%; p = 0.01). However, no influence of the graft material was noted in supragenicular bypasses (90.4% vs. 84.8%; p = 0.6). Bypasses in extremities with good runoff showed better patency than those in limbs showing poor runoff (86% vs. 55%; p = 0.003). The use of saphenous vein for the repair of popliteal aneurysms showed better results than those with prosthetic material, although in bypasses originating from the distal superficial femoral or above-knee popliteal artery, no significant differences in patency were observed. Good distal runoff was associated with improved overall outcome. PMID- 14564552 TI - The effect of gender on early and intermediate results of endovascular aneurysm repair. AB - Results of endovascular aneurysm repair (EVAR) may be gender dependent. Between September 1997 and September 2001, 118 AneuRx aortic grafts were placed for aneurysmal disease. During this period, 17 females and 101 males were treated with this device. A prospective database was maintained and supplemented with retrospectively gathered information to evaluate early and mid-term end points. A total of 113 devices were deployed in 118 attempts. Length of procedure was greater for females (3.3 +/- 1.75 vs. 2.3 +/- 0.8 hr, p = 0.05) and they were more likely to have significant arterial dissections (12% vs. 1%, p = 0.05). The mortality rates at 1 month were 12% for females and 0% for males ( p = 0.02); the complication rates at 1 month were 41% for females and 15% for males ( p = 0.02). Although technical success was not significantly different between the sexes, assisted primary technical success (requiring endovascular assistance) and assisted secondary technical success (requiring open surgical assistance) were significantly different (71% vs. 96%, p = 0.003; and 76% vs. 98%, p = 0.004, respectively). Clinical success at 1 month was 59% for females and 84% for males ( p = 0.02). This difference was also significant when assessing 1-month assisted primary clinical success (59% vs. 90%, p = 0.003) and assisted secondary clinical success as well (71% vs. 96%, p = 0.003). Clinical success and assisted primary clinical success were not different at 6- or 12-month intervals, however, assisted secondary clinical successes differed at both time intervals (56% vs. 83%, p = 0.02; and 56% vs. 81%, p = 0.05, respectively). As-yet undetermined factors appear to predispose females to complications and technical difficulties in the short term. Endovascular and open procedures required to achieve ongoing clinical success in the following months appear to favor males to a greater degree than females. PMID- 14564551 TI - Increased recognition of type II endoleaks using a modified intraoperative angiographic protocol: implications for intermittent endoleak and aneurysm expansion. AB - Retrograde arterial perfusion of the aneurysm sac (type II endoleak) may complicate endovascular abdominal aortic aneurysm (AAA) repair and may lead to AAA expansion and rupture. Aneurysm expansion may also occur in the absence of a demonstrable endoleak. Current intraoperative assessment techniques may underrepresent the incidence of type II endoleaks. This study evaluated the incidence and impact of previously unrecognized type II endoleaks using a modified intraoperative angiographic protocol. A total of 391 patients undergoing endovascular AAA repair were evaluated. In 264 patients standard completion angiograms were performed. In 127 patients a modified angiographic protocol was used to visualize collateral lumbar and inferior mesenteric arteries as well as the aneurysm sac. The modified protocol uses digital subtraction fluoroscopy continuously for 60 sec after injections of 20 mL iodinated contrast both in the pararenal aorta and within the endovascular graft. Postoperative CT scans were performed at 1, 6, and 12 months and annually thereafter. The average age was 73.3 years; 324 patients were men and 67 were women. Mean follow-up was 11.4 months (range, 1-60 months). Type II endoleaks were documented intraoperatively in a significantly increased proportion of patients in whom the modified angiographic protocol was used: modified, 53/127 = 41% vs. standard, 17/264 = 6%; p < 0.001. No significant difference in the incidence of type II endoleaks was present on CT scan at 6 or 12 months after surgery (6 months: modified, 6/72 = 8% vs. standard, 10/159 = 6%, p = NS; 12 months: modified, 2/36 = 5% vs. standard, 6/138 = 4%, p = NS). Forty-six type II endoleaks resolved spontaneously (10 in the standard cohort, 36 in the modified cohort). One patient had a 10-mm increase in AAA diameter after spontaneous thrombosis of a type II endoleak 18 months postoperatively. One patient had a type II endoleak intraoperatively and at 12 months after surgery but the endoleak was absent at 1 and 6 months. Thirteen patients from the standard protocol cohort and 1 from the modified protocol cohort developed newly visualized type II endoleaks during follow-up. These findings may imply intermittent patency of the artery supplying the type II endoleak. The overall morbidity rate was 14% and the perioperative mortality rate was 1.8%. Retrograde (type II) endoleaks originating from AAA side branches occur intraoperatively more frequently than is currently recognized. Intermittent patency and thrombosis of these vessels may also occur and may contribute to AAA expansion. The full significance of these previously unrecognized endoleaks with respect to risk of aneurysm rupture remains to be definitively determined. PMID- 14564553 TI - Iatrogenic hepatic artery pseudoaneurysms: an uncommon complication after hepatic, biliary, and pancreatic procedures. AB - Hepatic artery pseudoaneurysms are uncommon but potentially lethal complications of hepatic, biliary, and pancreatic interventions. To enhance our knowledge about these pseudoaneurysms, we reviewed our institution's experience with the management of these lesions. We reviewed the literature on 136 cases of hepatic artery pseudoaneurysms as well as our experience with 17 patients (excluding patients who were post-transplantation or had suffered abdominal trauma). The causes, pathogenesis, and clinical features were analyzed. Ten women and seven men developed hepatic artery pseudoaneurysms after undergoing hepatic (65%), biliary (30%), or pancreatic procedures (5%). The mean time between initial intervention and diagnosis was 5.7 months (range 7 days-38 months). Rupture occurred in 13 patients (76%). Mean pseudoaneurysm size was 1.9 cm (range 0.7-4 cm). Embolization was successful in 12 of 14 patients (86%). Four patients (24%), including the two who failed embolization, required operative intervention. Postoperative mortality was 25% while postembolization mortality was 14%. One patient was observed, and the aneurysm thrombosed at 72 months follow-up. Mean follow-up was 48 months (range 1-184 months) for 13 of the 14 survivors (93%) (1 patient was lost to follow-up) without any clinical sequela. Hepatic artery pseudoaneurysms are rare. Rupture is common and occurred in 76% of patients. For both ruptured and nonruptured cases angiography with embolization of the pseudoaneurysm is safe and effective. Operative intervention should be reserved for patients for whom embolization fails or for whom it is not feasible. PMID- 14564554 TI - Aortoiliac reconstruction: the retroperitoneal approach and splenic injury. AB - Exposure of the abdominal aorta through the left retroperitoneum is a well accepted technique. Unfortunately, splenic injury is a complication of this approach that rarely occurs through a mid-line transabdominal incision. In this series we evaluate the occurrence of splenic injury during left retroperitoneal aortic surgery. The records of all patients undergoing abdominal aortic reconstruction via the left retroperitoneal approach were reviewed from 1988 to 2001. Indications included either abdominal aortic aneurysm (AAA) or aortoiliac occlusive disease (AIOD). Thoracoabdominal aneurysms and visceral aortic reconstructions were excluded. Those patients that required splenectomy for splenic injury were stratified and analyzed for demographics, indications for operation, and other complications. Chi-square analysis was used to determine statistical significance. Over the 13-year study period, 2889 aortic reconstructions were performed. These consisted of 1773 elective AAA repairs, 357 ruptured or symptomatic AAA repairs, and 759 aortofemoral bypasses for AIOD. Splenectomies were performed after aortic repair in 21 (0.7%) patients. Breakdown by indication totaled 11/1773 (0.6%) for elective AAA repair, 7/357 (2.0%) for rupture or symptomatic AAA repair, and 3/759 (0.4%) for AIOD ( p < 0.05). Mortality for patients undergoing elective AAA without splenectomy was 1.9% (34/1762) and with splenectomy was 9.1% (1/11), while it was 3.2% (24/756) for AIOD patients without splenectomy and 14% (1/7) for AIOD patients with splenectomy (all p = NS). Splenic injury can occur in elective and emergent aortic reconstructions performed through the left retroperitoneum. The surgeon should be prepared for splenectomy whenever intraperitoneal blood or hemodynamic instability is identified. Evaluation of the spleen prior to flank closure may be prudent. PMID- 14564555 TI - Cytotoxic iron chelators: characterization of the structure, solution chemistry and redox activity of ligands and iron complexes of the di-2-pyridyl ketone isonicotinoyl hydrazone (HPKIH) analogues. AB - Di-2-pyridyl ketone isonicotinoyl hydrazone (HPKIH) and a range of its analogues comprise a series of monobasic acids that are capable of binding iron (Fe) as tridentate ( N, N, O) ligands. Recently, we have shown that these chelators are highly cytotoxic, but show selective activity against cancer cells. Particularly interesting was the fact that cytotoxicity of theHPKIH analogues is maintained even after complexation with Fe. To understand the potent anti-tumor activity of these compounds, we have fully characterized their chemical properties. This included examination of the solution chemistry and X-ray crystal structures of both the ligands and Fe complexes from this class and the ability of these complexes to mediate redox reactions. Potentiometric titrations demonstrated that all chelators are present predominantly in their charge-neutral form at physiological pH (7.4), allowing access across biological membranes. Keto-enol tautomerism of the ligands was identified, with the tautomers exhibiting distinctly different protonation constants. Interestingly, the chelators form low spin (diamagnetic) divalent Fe complexes in solution. The chelators form distorted octahedral complexes with Fe(II), with two tridentate ligands arranged in a meridional fashion. Electrochemistry of the Fe complexes in both aqueous and non-aqueous solutions revealed that the complexes are oxidized to their ferric form at relatively high potentials, but this oxidation is coupled to a rapid reaction with water to form a hydrated (carbinolamine) derivative, leading to irreversible electrochemistry. The Fe complexes of theHPKIH analogues caused marked DNA degradation in the presence of hydrogen peroxide. This observation confirms that Fe complexes from theHPKIH series mediate Fenton chemistry and do not repel DNA. Collectively, studies on the solution chemistry and structure of theseHPKIH analogues indicate that they can bind cellular Fe and enhance its redox activity, resulting in oxidative damage to vital biomolecules. PMID- 14564556 TI - Could the tyrosine-histidine ligand to CuB in cytochrome c oxidase be coordinatively labile? Implications from a quantum chemical model study of histidine substitutional lability and the effects of the covalent tyrosine histidine cross-link. AB - Density functional theory calculations have been used to evaluate the effects of inter-ring interactions within a covalently linked histidine-tyrosine cofactor such as that which is a ligand to the Cu(B) centre in cytochrome c oxidases and to investigate the energetics of histidine substitution at the Cu(B) centre. Small, but significant, perturbations of the redox potentials and/or p K(a) values of the histidine imidazole, the tyrosine phenol and the copper ion are found. The Cu(B)-N(cofactor) bond is estimated to be weaker than the Cu(B) N(histidine coligand) bonds in the Cu(B)(I) state and in the Cu(B) (II) state when the cofactor is oxidized, by approximately 13 kJ/mol and approximately 23 kJ/mol, respectively. The calculations reveal that displacement of a histidine ligand from the Cu(B) centre, as is suggested in proposals of "histidine cycle" mechanisms for proton pumping in cytochrome c oxidases, is only energetically feasible if accompanied by protonation of the histidine imidazole and coupled to an endothermic process. It is proposed that the histidine-tyrosine cofactor ought to be considered as the substitutionally labile ligand to Cu(B) as the covalent crosslink would ensure displacement of the cofactor from Cu(B)-driven helix deformation. It is estimated that this process could store up to approximately 70 kJ/mol, which, based upon thermodynamic considerations, is sufficient for the pumping of two protons in the later steps (reductive phase) of the catalytic cycle. Ramifications of this proposition for the mechanism of proton pumping in cytochrome c oxidases are discussed. PMID- 14564557 TI - Characterization of an Fe III-OOH species and its decomposition product in a bleomycin model system. AB - To model the mononuclear Fe(III)-OOH species identified in the catalytic cycle of the anticancer drug bleomycin, the iron chemistry of the pentadentate ligand N [bis(2-pyridylmethyl)aminoethyl]pyridine-2-carboxamide (H-PaPy(3)) has been investigated. The complex [Fe(III)(PaPy(3))OCH(3)](ClO(4)) was reacted with H(2)O(2) to form a red species (lambda(max)=480 nm, epsilon=1800 M(-1) cm(-1)) with an S=1/2 EPR signal at g=2.25, 2.17, and 1.95. This species has been identified by electrospray ionization mass spectrometry as [Fe(III)(PaPy(3))OOH](ClO(4)) and further characterized by resonance Raman and EXAFS analysis. The decomposition of this intermediate leads to the modification of the ligand, as revealed by (1)H NMR. One hydrogen atom is substituted by a solvent-derived methoxy group. The substitution at this site is a result of the two-electron oxidation of the ligand following the heterolytic cleavage of the O O bond of the Fe(III)-OOH species. This is a plausible mechanism to rationalize related ligand modifications that have been proposed in the decay of activated bleomycin. PMID- 14564558 TI - Comparative study of laparoscopic L5-S1 fusion versus open mini-ALIF, with a minimum 2-year follow-up. AB - Anterior lumbar interbody fusion (ALIF) is a widely accepted tool for management of painful degenerative disc disease. Recently, the modern laparoscopic surgical technique has been combined with ALIF procedure, with good early postoperative results being reported. However, the benefit of laparoscopic fusion is poorly defined compared with its open counterpart. This study aimed to compare perioperative parameters and minimum 2-year follow-up outcome for laparoscopic and open anterior surgical approach for L5-S1 fusion. The data of 54 consecutive patients who underwent anterior lumbar interbody fusion (ALIF) of L5-S1 from 1997 to 1999 were collected prospectively. More than 2-years' follow-up data were available for 47 of these patients. In all cases, carbon cage and autologous bone graft were used for fusion. Twenty-five patients underwent a laparoscopic procedure and 22 an open mini-ALIF. Three laparoscopic procedures were converted to open ones. For perioperative parameters only, the operative time was statistically different (P=0.001), while length of postoperative hospital stay and blood loss were not. The incidence of operative complications was three in the laparoscopic group and two in the open mini-ALIF group. After a follow-up period of at least 2 years, the two groups showed no statistical difference in pain, measured by visual analog scale, in the Oswestry Disability Index or in the Patient Satisfaction Index. The fusion rate was 91% in both groups. The laparoscopic ALIF for L5-S1 showed similar clinical and radiological outcome when compared with open mini-ALIF, but significant advantages were not identified, despite its technical difficulty. PMID- 14564560 TI - Effects of mycorrhizal colonisation on Thymus polytrichus from heavy-metal contaminated sites in northern England. AB - A study was performed to establish whether colonisation with arbuscular mycorrhizal (AM) fungi is beneficial to wild thyme [ Thymus polytrichus A. Kerner ex Borbas ssp. britannicus (Ronn.) Kerguelen (Lamiaceae)] growing in the heavy metal-contaminated soils along the River South Tyne, United Kingdom. T. polytrichus plants of the same genotype were grown under controlled conditions with and without Zn contamination, and differences between AM-colonised and uncolonised plants in mean shoot and root growth (dry weight) and Zn concentration were assessed. When grown in the heavy-metal-contaminated, low-P soil from one of the South Tyne sites, AM-colonised plants grew significantly larger than uncolonised plants; however, there was no significant difference in growth between AM and non-AM plants grown in an artificial substrate with a larger available P concentration, with or without Zn contamination. Mycorrhizal colonisation increased tissue Zn concentrations during the experiments. It is concluded that AM fungi are beneficial, if not essential, to T. polytrichus growing in the low-nutrient soils along the River South Tyne, because of their role in enhancing plant uptake of P (and possibly other nutrients). There was no evidence from this study that the fungi reduce plant uptake of heavy metals at these sites, but rather increase Zn uptake. However, the resulting tissue metal concentrations do not appear to be large enough to be detrimental to plant growth. PMID- 14564559 TI - Positive relationship between bone mineral density and low back pain in middle aged women. AB - There have been a large number of epidemiological studies demonstrating various primary factors that cause musculoskeletal disorders in middle-aged and older women. However, the relationship between low back pain and bone mineral density is not well documented, and no evidence for any direct relationship between the two has been found. To investigate the relationship, we conducted a cross sectional study, on a population of 2,244 Japanese women aged 25-85 years who were participating in a regional health screening program. Information on lifestyle, reproductive characteristics and the presence of current low back pain was collected by self-administered questionnaires, and bone mineral density at the distal radius was measured. We found increasing bone mineral density to be significantly associated with low back pain in middle-aged women using a logistic regression analysis. Exercise and smoking were also significantly associated with low back pain. This association remained even after entering other lifestyle and reproductive factors into the final model. Accordingly, high bone mineral density would seem to be as important a public health problem as low bone mineral density and osteoporosis when considering the musculoskeletal symptoms and disabilities that appear in middle-aged women. PMID- 14564561 TI - Changes in hemodynamic variables and catecholamine levels after rapid increase in sevoflurane or isoflurane concentration with or without nitrous oxide under endotracheal intubation. AB - PURPOSE: Rapid increases in concentrations of isoflurane and desflurane in oxygen have been shown to increase sympathetic activity. The aim of this study was to determine whether concomitant administration of nitrous oxide would reduce these sympathomimetic effects of volatile anesthetics. METHODS: Eighty healthy patients in whom the trachea was intubated and mechanically ventilated were given 15 min of anesthesia with either N(2)O (67%)-O(2)-sevoflurane (GOS), O(2)-sevoflurane (OS), N(2)O (67%)-O(2)-isoflurane (GOI), or O(2)-isoflurane (OI) (n = 20 per group). The inspired concentration of sevoflurane was 0.85% (0.5 minimum alveolar concentration [MAC]), and that of isoflurane was 0.6% (0.5 MAC). Fifteen minutes after endotracheal intubation, baseline and arterial blood sample data were obtained. Immediately after that, a sudden administration of 2.9 MAC volatile anesthetics was performed. Systolic and diastolic arterial pressures, heart rate, and end-tidal carbon dioxide concentration were obtained at 0.5, 1, 1.5, 2, 3, 4, and 5 min after that. To measure catecholamine levels, arterial blood samples were obtained 2 and 5 min after the trial started. RESULTS: Except for the OI group, systolic and diastolic arterial pressure progressively decreased after the abrupt increase in the concentration of volatile anesthetics. Except for the OS group, the heart rate increased after the abrupt increase in the concentration of volatile anesthetics. In the OI group, the end-tidal concentration of carbon dioxide increased at 0.5 and 1 min, suggesting that a slight hyperdynamic state occurred. However, it decreased progressively after the abrupt increase in volatile anesthetic concentration in the other groups. Plasma norepinephrine levels increased progressively in all groups. CONCLUSION: Even if nitrous oxide was added to isoflurane or sevoflurane, the increase in heart rate could not be avoided. Contrary to previous reports, severe hyperdynamic circulation was not observed after a rapid increase in isoflurane concentration. PMID- 14564562 TI - Unanswered and uncounted items of the illness behavior questionnaire count: a comparison between patients who answered all the items in the questionnaire and those who left one or more items unanswered in a pain management program. AB - PURPOSE: This study aimed to understand the significance of unanswered and uncounted items on the illness behavior questionnaire (IBQ) in the setting of the Mayo Clinic pain management program for patients with chronic pain. METHODS: Three hundred and seventy-eight patients who completed the questionnaire were studied. The data included (1) age, (2) IQs, (3) IBQ score profiles, (4) litigation status, (5) admission status (inpatient vs outpatient), and (6) dismissal status ("graduates" vs "dropouts"). RESULTS: Comparison between patients who answered all the items (n = 272) and those who left one or more items unanswered (n = 106) showed a significant difference in the completion rate of the Wechsler Adult Intelligence Scale ( P < 0.05), Scale 5 (affective disturbance) score (P < 0.05), and the rate of dropout from the program (P < 0.05). Also, those who dropped out of the pain management program had a significantly lower score on Scale 5 (P < 0.001) and a significantly greater number of unanswered items that were not counted toward the scale score (P < 0.05). CONCLUSION: Unanswered and uncounted items of the IBQ seem to count toward a better understanding of patients' pain behavior. PMID- 14564563 TI - Pentobarbital and ketamine suppress serum concentrations of sex hormones in the female rat. AB - PURPOSE: To investigate the potential effects of pentobarbital and ketamine on serum concentrations of sex hormones, the present study was performed using pregnant mare serum gonadotropin (PMSG)-primed cyclic female Sprague-Dawley rats. METHODS: Pentobarbital sodium (37 mg.kg(-1), i. p.) or ketamine-hydrochloride (229 mg.kg(-1), i. m.) was injected 2 and 3 days after PMSG treatment. At 0, 1, 2, 3, 4, and 5 days after PMSG treatment, sera were collected by cardiac puncture. The serum concentrations of progesterone (P(4)), testosterone (T), and estradiol-17 beta (E(2)) were determined by radioimmunoassay. RESULTS: The serum concentrations of P(4) tended to be lower in both the pentobarbital- and the ketamine-treated groups compared with the control group. Significant differences were found on days 3 and 4 after pentobarbital and on days, 1, 4, and 5 after ketamine administration. Serum concentrations of T were also suppressed in both the pentobarbital and the ketamine-treated groups, whereas E(2) concentrations decreased only in the ketamine-treated group. CONCLUSION: Pentobarbital and ketamine decrease serum sex hormone concentrations in PMSG-primed female rats. PMID- 14564564 TI - A comparison between low-flow leak test and oxygen flush leak test. AB - PURPOSE: The clinical efficacy of two methods of preanesthetic leak test,namely the oxygen flush leak test (OFLT) and the low-flow leak test (LFLT), was compared regarding their ability to detect leakage in the anesthesia circuit and their accuracy. METHODS: Examinees comprised 16 staff anesthesiologists and 7 physicians undergoing anesthesia training at our institution. They performed the two leak tests on anesthesia machines with some intentional leaks (0.1-1.0 l.min( 1)). The leakage detection rates (LDR) were analyzed by the chi(2)-test. The ability to detect leaks was measured by recording how many leaks were detected by 50% (LDR(50)) and 95% (LDR(95)) of the examinees. RESULTS: The LDRs in the two tests were significantly different (chi(2)- analysis, P < 0.0001). Both LDR(50) and LDR(95) for the LFLT (0.23 and 0.41 l.min(-1), respectively) were smaller than the values for the OFLT (0.37 and 0.82 l.min(-1), respectively). The sensitivity and specificity of the LFLT (0.97 and 0.84, respectively) were higher than those of the OFLT (0.78 and 0.80, respectively). CONCLUSION: The LFLT was found to be superior to the OFLT regarding leak detection and reliability. We therefore recommend the LFLT for preanesthetic leak testing. PMID- 14564565 TI - The combination of hemodilution and controlled hypotension: physiology and clinical application. PMID- 14564566 TI - Ruptured superior mesenteric artery aneurysms during treatment of tetanus: a case report. PMID- 14564567 TI - Variant form of stiff-man syndrome with neck pain: report of a case treated with muscle afferent block. PMID- 14564568 TI - Anesthesia for correction of esophageal stricture in a patient with epidermolysis bullosa: a case report. PMID- 14564569 TI - Anesthetic management of a child with Rubinstein-Taybi syndrome for cervical dermoid cyst excision. PMID- 14564571 TI - Subarachnoid venous hemorrhage in a patient with retrograde cerebral perfusion during surgery for a thoracic aortic aneurysm. PMID- 14564570 TI - Postoperative left recurrent laryngeal nerve palsy possibly caused by coincidental swelling of the metastatic mediastinal lymph node. PMID- 14564572 TI - Gastric tube guide-equipped laryngeal mask airway. PMID- 14564573 TI - Estimation of spread of lumbar or lower thoracic epidural anesthesia by a simple equation. PMID- 14564574 TI - A Bullard laryngoscope holder for bedside use. PMID- 14564575 TI - Limited spread of spinal anesthesia using a hyperbaric local anesthetic in older patients. PMID- 14564576 TI - Platelet function in pregnant women receiving aspirin and dipyridamole. AB - PURPOSE: This study was undertaken to evaluate the hemostasis and coagulation profile of pregnant women receiving antiplatelet therapy with low-dose aspirin and dipyridamole for prevention of preeclampsia, intrauterine growth retardation, or pregnancy losses. METHODS: Twenty-three pregnant women who received antiplatelet therapy with combined aspirin (40 mg.day(-1)) and dipyridamole (150 mg.day(-1)) were enrolled in the study. Platelet aggregation and coagulation tests were performed before the start of aspirin and dipyridamole, during medication, and at 3 days and 6 days after cessation of medication. RESULTS: Collagen-induced platelet aggregation was decreased during medication (25 +/- 26%, P < 0.001) and at 3 days after cessation of medication (46 +/- 35%, P < 0.001) compared with that before the start of medication (89 +/- 7%). ADP-induced platelet aggregation was decreased during medication compared with that before medication (66 +/- 18% vs 92 +/- 7%, P < 0.001). The platelet count, prothrombin time, activated partial thromboplastin time, bleeding time, and levels of fibrinogen and antithrombin III did not change over time. The blood loss of these patients during vaginal delivery and cesarean section did not differ from that of normal women during vaginal delivery and repeat cesarean section, respectively. CONCLUSION: At the doses used in this study, aspirin and dipyridamole inhibited platelet aggregation for up to 3 days after cessation of medication. This abnormality of aggregation was not detected by the bleeding time and was not associated with clinically abnormal bleeding. PMID- 14564577 TI - Continuous intra-arterial blood gas monitoring during thoracic surgery. AB - PURPOSE: This study was undertaken to assess the clinical usefulness of a continuous intra-arterial blood gas (CIABG) monitoring system, Paratrend 7, during thoracic surgery. METHODS: A sensor of the CIABG monitoring system was inserted into the radial artery in 50 patients. During one-lung/differential lung ventilation, arterial blood samples for estimation of blood gases (ABG) were taken every 45-90 min or when clinically needed. The PCO(2), PO(2), and pH values displayed by the CIABG monitor were recorded prior to arterial blood sampling and were compared with the results of ABG analysis. The mean (bias) and the standard deviation (precision) of the differences were calculated from the data for each parameter. RESULTS: Ninety-four blood samples were obtained. The correlation between CIABG and ABG measurements was strong for each parameter: r = 0.83 (PCO(2)), 0.89 (PO(2)), 0.74 (pH). The bias +/- precision between the two methods was 0.4 +/- 3.0 mmHg for PCO(2), -6 +/- 47 mmHg for PO(2), -1.2 +/- 27.4% for PO(2) and -0.01 +/- 0.04 for pH. For PO(2) values <150 mmHg, the bias +/- precision was 1 +/- 28 mmHg. CONCLUSION: The agreement between CIABG and ABG measurements was better for PCO(2) and pH than for PO(2). The PO(2) value displayed on the CIABG monitor may not always be reliable during thoracic surgical procedures. PMID- 14564578 TI - Pharmacokinetic profile of propofol after a single-dose injection during general anesthesia in Japanese adults. AB - PURPOSE: To determine the pharmacokinetic parameters of propofol after a single dose injection in Japanese adults. METHODS: This study was carried out in adult patients who underwent minor surgery under general anesthesia with sevoflurane. We injected 1.0, 1.5, or 2.0 mg.kg(-1) of propofol at a constant rate using a syringe pump. Arterial blood samples were taken for 480 min after the administration of propofol. The whole-blood concentration of propofol was determined with gas chromatography, and a time-blood concentration curve was analyzed by a two-compartment open-model analysis and a model-independent analysis. RESULTS: The half-lives of the central and peripheral compartment ( t(1/2)alpha and t(1/2)beta) were 2.26 +/- 0.69 and 47.9 +/- 22.1 min, respectively. The volume of the central compartment (V(c)) was 0.582 +/- 0.170 l.kg(-1), and the apparent volume of distribution at a steady state (V(dss)) was 2.62 +/- 1.06 l.kg(-1). The total body clearance (Cl) and mean residence time (MRT) were 53.7 +/- 11.9 ml.min(-1).kg(-1) and 98.1 +/- 16.4 min, respectively. CONCLUSIONS: Among the pharmacokinetic parameters determined in Japanese adults, t(1/2)alpha, t(1/2)beta, and Vc were similar, V(dss) was smaller, and Cl was larger, as compared with values in Caucasians. These findings suggest that propofol could be eliminated well during minor surgery in Japanese adults. PMID- 14564579 TI - Propofol injection pain is not alleviated by pretreatment with flurbiprofen axetil, a prodrug of a nonsteroidal antiinflammatory drug. AB - PURPOSE: The effects of nonsteroidal antiinflammatory drugs (NSAIDs) on pain from propofol injection are controversial, partially because NSAIDs themselves cause injection pain. We evaluated the effects of flurbiprofen axetil (LFP), a prodrug of an NSAID, on pain induced by intravenous propofol injection, because LFP produces little pain on injection. METHODS: A randomized, double-blind, controlled trial was undertaken in patients who were assigned to one of three groups ( n = 50 in each). Patients received either 5 ml of saline followed approximately 10 min later by propofol mixed with 0.4 ml of saline, LFP (50 mg, 5 ml) i.v. followed by propofol mixed with 0.4 ml of saline, or 5 ml of saline followed by propofol mixed with lidocaine (40 mg, 0.4 ml). Verbal rating scores for injection pain were assessed every 10 s during propofol administration at a rate of 0.05 mg.kg(-1).s(-1). RESULTS: None of the patients complained of pain during injection of LFP or saline. Admixture of lidocaine, but not of LFP, significantly reduced the incidence of pain and the severity of pain scores during propofol injection ( P = 0.0017 and P < 0.001, respectively). CONCLUSION: Lidocaine, but not LFP, is effective for controlling pain induced by propofol injection. This result suggests that NSAIDs have little effect on pain from propofol injection. PMID- 14564580 TI - Changes in left ventricular end-diastolic area, end-systolic wall stress, and fractional area change during anesthetic induction with propofol or thiamylal. AB - PURPOSE: To elucidate the mechanisms of the more profound hypotensive effects of propofol relative to thiamylal, we monitored changes in left ventricular (LV) preload, afterload, and contractility during the course of anesthetic induction with propofol and thiamylal. METHODS: Thirty-two patients (ASA I) were randomly assigned into two groups and injected with propofol (2 mg.kg(-1)) or thiamylal (4 mg.kg(-1)) as anesthetic induction agents. Transthoracic echocardiography (TTE) was used to assess LV performance before and during induction by the two anesthetics. The LV end-diastolic area (EDA) and LV end-systolic wall stress (ESWS) were used as indices of LV preload and LV afterload, respectively, while LV contractility was assessed by the fractional area change (FAC). RESULTS: Both propofol and thiamylal significantly reduced EDA and ESWS without significant change in FAC. Propofol-induced reductions in EDA and ESWS were significantly greater than those of thiamylal. CONCLUSION: The more profound hypotension observed during induction of anesthesia with propofol is due to the greater decrease in preload and afterload than with thiamylal, but not to a decrease in LV contractility. PMID- 14564581 TI - Quantification of the degradation products of sevoflurane using four brands of CO2 absorbent in a standard anesthetic circuit. AB - PURPOSE: CO(2) absorbents convert sevoflurane to fluoromethyl-2,2-difluoro-1 (trifluoromethyl) vinyl ether (compound A), whose toxicity in rats raises concern regarding the safety of sevoflurane in a low-flow system. The type of CO(2) absorbent is one of factors that affect compound A concentration in the anesthetic circuit. The aim of the present study was to investigate the concentration of compound A in an anesthetic model circuit following the use of different brands of soda lime and Baralyme. METHODS: We measured the concentrations of compound A in four different brands of CO(2) absorbent using a low-flow (1 l.min(-1) fresh gas) model circuit in which 2% sevoflurane was circulated. Sodasorb II, Baralyme, Sofnolime and Wakolime-A were used as CO(2) absorbents. The concentration of compound A was measured hourly, and the temperature of the CO(2) absorbent was monitored. RESULTS: The maximum concentration of compound A in the circuit was highest for Baralyme (25.5 +/- 0.6 ppm) (mean +/- SD), followed by Sodasorb II (18.9 +/- 1.6 ppm), Wakolime-A (16.1 +/- 0.7 ppm), and Sofnolime (15.8 +/- 1.4 ppm). The maximum temperature was 50.8 +/- 1.3 degrees C for Baralyme, 48.8 +/- 1.3 degrees C for Wakolime-A, 47.0 +/- 1.4 degrees C for Sodasorb II, and 43.5 +/- 3.9 degrees C for Sofnolime. CONCLUSION: The relative concentrations of compound A in the low-flow circuit were Baralyme > Sodasorb II > Wakolime-A = Sofnolime. PMID- 14564582 TI - Effect of linear polarized light radiation on impaired mitochondrial oxidative phosphorylation in skeletal muscle. AB - PURPOSE: The aim of this study was to investigate the effect of linear polarized light radiation (LPLR) on mitochondrial oxidative phosphorylation impaired by hemorrhagic shock or Escherichia coli lipopolysaccharide (LPS) in skeletal muscle. METHODS: We studied the effect of LPLR on mitochondrial function of skeletal muscle by using a model of mitochondria impaired by hemorrhage or LPS. The oxygen uptake in states 3 and 4, the respiratory control ratio (RCR), and the adenosine diphosphate-to-oxygen ratio (ADP/O) were measured with a Clark oxygen electrode. RESULTS: Oxygen uptake in states 3 and 4, RCR, and ADP/O were significantly decreased by hemorrhage for 4 h and by LPS treatment for 12 h. Oxygen uptake in states 3 and 4 impaired by hemorrhage increased significantly from 40.1 +/- 3.2 to 60.1 +/- 5.4 nmol O(2).min(-1).mg protein(-1) after LPLR, and oxygen uptake in state 4 decreased significantly from 22.8 +/- 2.4 to 17.7 +/ 1.5 nmol O(2).min(-1).mg protein(-1) after LPLR. RCR and ADP/O were also significantly increased from 1.8 +/- 0.3 and 0.9 +/- 0.2 to 3.4 +/- 0.3 and 1.5 +/- 0.1, respectively, by LPLR. Oxygen uptake in states 3 and 4 impaired by LPS was improved from 46.6 +/- 5.1 and 21.0 +/- 1.9 to 53.8 +/- 6.2 and 17.9 +/- 2.3 nmol O(2).min(-1).mg protein(-1), respectively following LPLR. RCR and ADP/O were also elevated significantly from 2.2 +/- 0.2 and 0.9 +/- 0.2 to 3.0 +/- 0.3 and 1.4 +/- 0.2, respectively, after LPLR. CONCLUSION: LPLR improved mitochondrial oxidative phosphorylation of skeletal muscle impaired by hemorrhagic shock or E. coli LPS. PMID- 14564583 TI - Myocardial ischemia due to paradoxical air embolism during living-related donor liver transplantation in a child. PMID- 14564584 TI - Serum mucin-like glycoprotein antigen (KL-6) as a specific marker of interstitial pneumonia: a case report. PMID- 14564585 TI - Pulmonary edema after laparoscopic adrenalectomy in a pregnant patient with Cushing's syndrome. PMID- 14564586 TI - Unexpected resistance to pancuronium in a patient with myotonic dystrophy (myotonia dystrophica). PMID- 14564587 TI - A randomized, double-blind trial comparing the effect of mixing propofol with either lidocaine or nafamostat mesilate on injection pain. PMID- 14564588 TI - Binding of [14C]thiopental to rat cerebrocortical membranes. PMID- 14564589 TI - Inadvertent subcutaneous infusion of propofol by a syringe pump. PMID- 14564590 TI - pH-Stat management is more appropriate during deep hypothermia, especially when circulatory arrest is added. PMID- 14564591 TI - Effects of sevoflurane anesthesia combined with epidural block on renal function in the elderly: comparison with isoflurane. AB - PURPOSE: Renal function declines with age, but little is known about the renal effects of the inhaled anesthetic sevoflurane in the elderly. We therefore compared the renal effects of sevoflurane and isoflurane anesthesia in elderly patients. METHODS: Thirteen patients aged > or =70 years undergoing gastrectomy with epidural anesthesia combined with general anesthesia were randomly assigned to receive either sevoflurane ( n = 7) or isoflurane ( n = 6). Dopamine (3-5 microg.kg(-1).min(-1)) was administered to all patients. Blood and urine samples were collected before, during, and after anesthesia. Serum and urinary inorganic fluoride was measured, and renal function tests were performed. RESULTS: Serum inorganic fluoride was significantly elevated in both groups. The production of inorganic fluoride was significantly greater in the sevoflurane group, but the level did not exceed 50 micromol.l(-1) in any patient. No abnormalities were observed in blood urea nitrogen (BUN), serum creatinine, or urine volume in either group. The albumin excretion index increased during anesthesia and decreased after anesthesia in both groups. Creatinine clearance was unchanged in the sevoflurane group but fluctuated during and after anesthesia in the isoflurane group. alpha(1)-Microglobulin (MG), beta(2)-MG, and urinary N-acetyl beta-D-glucosaminidase (NAG) excretion increased up to 3 h after anesthesia, and alpha(1)-MG and beta(2)-MG excretion increased on postanesthesia day 3. CONCLUSION: In both groups, glomerular and tubular function were transiently affected, but no abnormalities were found in routine laboratory tests, suggesting that neither isoflurane nor sevoflurane in combination with dopamine and epidural anesthesia seriously affects renal function in the elderly. PMID- 14564592 TI - An induction dose of propofol does not alter cerebral blood flow determined by single-photon-emission computed tomography. AB - PURPOSE: To examine the effect of propofol per se on cerebral blood flow (CBF), we measured CBF by single-photon-emission computed tomography (SPECT) using a technetium-99m ethyl cysteinate dimer before and after propofol administration. METHODS: Ten healthy adult male volunteers were studied. Ten minutes after isotope injection, CBF was measured using a SPECT system. After this first SPECT scan, 1.5 mg.kg(-1) of 1% propofol was administered over 30 s and the same dose of isotope was injected 2 min thereafter. Ten minutes later, a second SPECT scan was carried out. A subtraction SPECT image was obtained by reducing the first SPECT image from the second SPECT image. Based on these SPECT images, various regions of interest (ROI) were traced. Changes in regional CBF to each ROI were analyzed by a comparison of the total gamma-ray counts in each ROI between the first and subtraction SPECT images. RESULTS: The total gamma-ray counts in each ROI did not change significantly after propofol administration. At this time, end tidal carbon dioxide concentration and heart rate did not change, but blood pressure and oxygen saturation decreased slightly. CONCLUSION: The present result suggests that the induction dose of propofol does not alter CBF. PMID- 14564593 TI - Appropriate method of administration of propofol, fentanyl, and ketamine for patient-controlled sedation and analgesia during extracorporeal shock-wave lithotripsy. AB - PURPOSE: The aim of this study was to identify the appropriate method for administering propofol, fentanyl, and ketamine (PFK) for patient-controlled sedation and analgesia (PCSA) during extracorporeal shock-wave lithotripsy (ESWL). METHODS: Twenty-one unpremedicated patients were randomly assigned to three groups that received different drug administration regimens. (group 1: low loading dose and high demand bolus, group 2: high loading dose and demand bolus, group 3: high loading dose and low demand bolus). RESULTS: The patients in all groups were hemodynamically stable during ESWL. Oxygen desaturation was recognized in all groups, but was avoided by 2 l.min(-1) of oxygen supply via a nasal prong. The total administration dose of the drugs was significantly higher ( P < 0.05) in group 2 than in groups 1 and 3. The median level of sedation was the same, but the episodes of oversedation were not recognized in group 3 ( P < 0.05). A significant difference in the frequency of episodes of oversedation was found between groups 2 and 3 ( P < 0.05). The results were good or excellent for almost all patients, and were assessed as fair by only one patient in group 2. CONCLUSION: We concluded that the method used for group 3 is the most appropriate for administering PFK for PCSA during ESWL. PMID- 14564594 TI - Reinforcement of spinal anesthesia by epidural injection of saline: a comparison of hyperbaric and isobaric tetracaine. AB - PURPOSE: An epidural injection of saline was reported to extend spinal anesthesia because of a volume effect. The aim of this study was to evaluate the influence of the baricity of spinal local anesthetics upon the extension of spinal anesthesia by epidural injection of saline. METHODS: Forty patients undergoing elective lower-limb surgery were randomly allocated to four groups of 10 patients each. Group A received no epidural injection after the spinal administration of hyperbaric tetracaine (dissolved in 10% glucose). Group B received an epidural injection of 8 ml of physiological saline 20 min after spinal hyperbaric tetracaine. Group C received no epidural injection after spinal isobaric tetracaine (dissolved in physiological saline). Group D received an epidural injection of 8 ml of saline 20 min after spinal isobaric tetracaine. The level of analgesia was examined by the pinprick method at 5-min intervals. RESULTS: The levels of analgesia 20 min after spinal anesthesia were significantly higher in hyperbaric groups than in isobaric groups [T5 (T2-L2) vs. T7 (T3-12)]. After epidural injection of saline, the levels of analgesia in groups B and D were significantly higher than in groups A and C. The segmental increases after epidural saline injection were 2 (0-3) in group B and 2 (1-7) in group D. Sensation in the sacral area remained 20 min after spinal block in one patient in group D; however, it disappeared after epidural saline injection. CONCLUSION: In this study, 8 ml of epidural saline extended spinal analgesia. However, there was no difference between the augmenting effect in isobaric and hyperbaric spinal anesthesia. We conclude that the reinforcement of spinal anesthesia by epidural injection of saline is not affected by the baricity of the spinal anesthetic solution used. PMID- 14564595 TI - Nicardipine versus lidocaine for attenuating the cardiovascular response to endotracheal intubation. AB - PURPOSE: The aim of this study was to compare the efficacy of nicardipine and lidocaine in attenuation of cardiovascular responses to endotracheal intubation. METHODS: In a randomized, double-blind, controlled trial, 60 unpremedicated (ASA I) patients undergoing elective surgery were given either 30 microg.kg(-1) nicardipine or 1.5 mg.kg(-1) lidocaine intravenously 2 min before intubation. Laryngoscopy and tracheal intubation were performed 1 min after induction of anesthesia with 5 mg.kg(-1) thiopentone, followed by administration of 1.5 mg.kg( 1) succinylcholine intravenously. Blood pressure and heart rate were monitored at baseline and every minute until 4 min after intubation. Repeated-measures ANOVA, Student's t test, the chi-square test, and 95% confidence intervals were used as appropriate. P < 0.05 was considered statistically significant. RESULTS: Baseline hemodynamic variables were not different between the groups. After administration of either agents, diastolic blood pressure and mean blood pressure were significantly lower in the nicardipine group. The heart rate in the nicardipine group was significantly higher. The mean between-group differences in diastolic blood pressure, mean blood pressure, heart rate, and rate-pressure product at baseline and 1 min after starting laryngoscopy were statistically significant. CONCLUSION: Nicardipine can be used as an alternative to lidocaine in attenuation of cardiovascular response to tracheal intubation in patients without ischemic heart disease. PMID- 14564597 TI - Managing perioperative hypothermia. PMID- 14564598 TI - Anesthetic management during minimally invasive cardiac surgery with the Port Access system for closure of atrial septal defect. PMID- 14564596 TI - Halothane anesthesia decreases the extracellular level of dopamine in rat striatum: a microdialysis study in vivo. AB - PURPOSE: In our previous microdialysis study, sevoflurane or isoflurane anesthesia significantly decreased the extracellular level of dopamine in rat striatum in vivo. On the other hand, other investigators demonstrated that halothane anesthesia either increased or did not affect the extracellular dopamine level. To explore the differences among these volatile anesthetics, the effects of halothane and nitrous oxide on the striatal dopamine level were reinvestigated. METHODS: Halothane alone, nitrous oxide with or without halothane, or drugs known to affect the dopaminergic pathway were administered to rats. Microdialysates were collected every 20 min and directly applied to an on line high-performance liquid chromatograph without any pretreatment. The effects of halothane on respiratory and cardiovascular variables were monitored. RESULTS: General anesthesia with halothane alone decreased the dialysate (extracellular) concentration of dopamine but increased that of dopamine metabolites. Nitrous oxide alone slightly increased dopamine metabolites in dialysates but did not affect the halothane-induced decrease in extracellular dopamine. Apomorphine and haloperidol reproduced reported results, confirming the adequacy of our methodology. Nomifensine- or methamphetamine-induced increase in extracellular dopamine was augmented by halothane. CONCLUSION: These results suggest that halothane potently enhances striatal dopamine release and activates the reuptake or metabolic process, which is consistent with our previous results for sevoflurane or isoflurane. Volatile anesthetics interfere with dopamine regulation, at least in the rat striatum. PMID- 14564599 TI - Undiagnosed sick sinus syndrome manifest during combined general and cervical epidural anesthesia. PMID- 14564600 TI - Perioperative endocrinological findings in a patient with Bartter's syndrome and living-related renal transplantation. PMID- 14564601 TI - Anesthetic management of heterotopic heart transplantation on beating heart. PMID- 14564602 TI - Effects of sevoflurane compared with those of isoflurane on arterial oxygenation and hemodynamics during one-lung ventilation. AB - PURPOSE: This study was designed to compare the effects of sevoflurane and isoflurane on Pao(2) and hemodynamic variables during one-lung ventilation (OLV) in surgical patients. METHODS: Twelve patients undergoing an esophageal procedure with thoracotomy for which a long period of OLV was required were studied using a randomized crossover design. Group 1 received 1.2% isoflurane from the induction of anesthesia until 30 min after starting OLV, and then received 1.7% sevoflurane during the remaining period. In group 2, the order of the anesthetics was reversed. All experimental procedures were performed in the left lateral decubitus position with the chest opened. Arterial and mixed venous blood gases and cardiac outputs were analyzed immediately before OLV, during OLV, and after resumption of two-lung ventilation (TLV). RESULTS: OLV produced lower PaO(2) and higher venous admixture (Q(s)/Q(t)) values than TLV. However, there was no significant difference between sevoflurane and isoflurane in PaO(2) or Q(s)/Q(t) during OLV. Other hemodynamic variables except for PVO(2) showed no significant differences between the anesthetics. CONCLUSION: The effects of sevoflurane on PaO(2) and the hemodynamic variables were similar to those of isoflurane during TLV and OLV in the lateral decubitus position. PMID- 14564603 TI - Comparison of the in vitro caffeine-halothane contracture test with the Ca induced Ca release rate test in patients suspected of having malignant hyperthermia susceptibility. AB - PURPOSE: We compared the results of the in vitro caffeine-halothane contracture test (CHCT) according to the protocols of the North American Malignant Hyperthermia Group (NAMHG) and the European Malignant Hyperthermia Group (EMHG) with the Ca-induced Ca release (CICR) rate test in the same patients with suspected malignant hyperthermia (MH). METHODS: Five normal controls and 16 patients suspected of having MH susceptibility were studied. Muscle biopsies were usually obtained from the musculus vastus lateralis. Diagnostic cutoff points and procedures for CHCT protocols were as described in the original and renewal versions of NAMHG and EMHGs. The CICR rate test was performed according to the protocol reported by Endo et al. RESULTS: All five normal controls and two patients with abortive MH, two with postoperative hyperthermia, and three with high serum creatine kinase levels were normal in the three tests. Three patients with MH reactions and one patient with a history of masseter spasm were classified as MH positive according to NAMHG criteria and MH susceptible and MH equivocal according to EMHG criteria. There were five cases with discordant results between the CHCT and CICR rate tests. CONCLUSION: We propose that muscle biopsy for diagnosis of MH susceptibility should combine the CHCT with the CICR rate test, which may identify the defective site of Ca release channels. PMID- 14564604 TI - What is the optimal dose of glucose administration during minor surgery under sevoflurane anesthesia? AB - PURPOSE: We attempted to identify the optimal infusion rate of glucose to maintain an appropriate usage of energy sources during minor surgery after an overnight fast. METHODS: Forty patients scheduled for tympanoplasty or skin grafting under sevoflurane anesthesia were assigned to four groups. The patients received a 2-h infusion of either saline or glucose at a rate of 0.1, 0.2, or 0.3 g.kg(-1).h(-1). Blood samples were collected before the induction of anesthesia, and at 1 and 2 h after the start of the saline or glucose infusion. Plasma glucose, free fatty acid, beta-hydroxybutyrate, acetoacetate, and immunoreactive insulin were measured. RESULTS: Plasma glucose concentration increased dose dependently. Immunoreactive insulin levels increased in the groups receiving 0.2 or 0.3 g.kg(-1).h(-1) of glucose infusion. Free fatty acid and ketone bodies did not increase in any glucose infusion groups. The arterial ketone body ratio increased to over 1.00 in the groups receiving 0.2 or 0.3 g.kg(-1).h(-1) of glucose infusion. Glycorrhea was observed only in the group receiving 0.3 g.kg( 1).h(-1) of glucose. CONCLUSION: The smaller doses of glucose (0.1-0.2 g.kg( 1).h(-1)) prevented lipolysis and hyperglycemia during minor surgery. PMID- 14564606 TI - Effects of nitric oxide synthase inhibition on extracellular glutamate and cerebral blood flow during forebrain ischemia-reperfusion in rat in vivo. AB - PURPOSE: To evaluate factors involved in global forebrain ischemia-reperfusion, the effects of the systemically administered NOS inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME), on changes in extracellular glutamate and cerebral blood flow (CBF) were studied during the early period of global forebrain ischemia reperfusion, simultaneously measuring the glutamate released in the rat forebrain cortex and cortical CBF. METHODS: After injection of saline or L-NAME, forebrain ischemia-reperfusion was performed by bilateral carotid artery occlusion with controlled hemorrhagic hypotension (30 mmHg) for 10 min and reperfusion for 60 min. The microdialysis electrode and laser Doppler flowmetry were used for real time monitoring of glutamate and CBF, respectively. RESULTS: During ischemia, glutamate increased linearly to over 100 muM and remained elevated 30 min after reperfusion in L-NAME-treated rats. In L-NAME-treated rats, CBF also remained significantly lower than baseline for 30-60 min after reperfusion, and glutamate was higher than in saline-treated rats throughout the experiment. CONCLUSION: A remarkable linear increase in glutamate release was observed during ischemia. L NAME did not prevent this dramatic glutamate accumulation, and moreover, its level increased during reperfusion. The decrease in CBF response after reperfusion might be a factor of the elevated glutamate after reperfusion due to a decrease in reuptake of glutamate. PMID- 14564605 TI - Optimal dose and duration of glucose administration during fasting period preceding surgery in rabbits. AB - PURPOSE: We examined preoperative glucose administration to establish what dose and cutoff point were optimal for suppression of lipolysis and prevention of hypo or hyperglycemia. METHODS: Rabbits were preoperatively fasted and simultaneously received glucose at a constant rate of 0, 0.1, 0.2, 0.3, or 0.4 g.kg(-1).h(-1) in fluid infusion for 3 h. Plasma glucose, immunoreactive insulin activity, nonesterified fatty acids, and ketone bodies were measured 0, 1.5, 3 and 4 h after the start of infusion, and hepatic glycogen content was assessed 1 h after cessation of infusion. RESULTS: Fluid infusion without glucose decreased plasma glucose. Glucose administration at more than 0.2 g.kg(-1).h(-1) caused hyperglycemia (>200 mg.dl(-1)) in the infusion period; the differences were significant compared with the value at zero time or in the 0 g.kg(-1).h(-1) group (P < 0.01). The highest dose also raised plasma immunoreactive insulin activity, which was significantly higher than in the 0 g.kg(-1).h(-1) group (P < 0.01) at the midpoint of the infusion period. Plasma nonesterified fatty acids increased in all groups. The changes were, however, significantly reduced in both the 0.3 and 0.4 g.kg(-1).h(-1) groups (P < 0.05 and P < 0.01, respectively) by the end of infusion. All these effects of glucose supply, including suppression of lipolysis, disappeared regardless of dose within 1 h after the cessation of infusion. CONCLUSION: These results suggest that the optimal dose for preoperative glucose infusion, in order to preserve carbohydrate or fat metabolism, is 0.1-0.2 or 0.3 g.kg(-1).h(-1), respectively, and indicate that administration should not be discontinued until the start of surgery. PMID- 14564607 TI - Inhibition of human plasma cholinesterase and erythrocyte acetylcholinesterase by nondepolarizing neuromuscular blocking agents. AB - PURPOSE: The kinetics of the inhibition of human plasma cholinesterase (ChE) and erythrocyte acetylcholinesterase (AChE) by alcuronium, atracurium, d tubocurarine, pancuronium, pipecuronium, and vecuronium were studied in blood drawn from 35 surgical patients. METHODS: The activities of plasma ChE and erythrocyte AChE were determined by the calorimetric method of Ellman et al., using acetylthiocholine as the substrate. Lineweaver-Burk plots and Dixon plots were used for the analysis of the kinetics of both enzymes. RESULTS: The dissociation constants (K(m)) of plasma ChE and erythrocyte AChE were 5.00 x 10( 5) M and 5.28 x 10(-5) M, respectively, indicating that both enzymes have similar affinity to acetylthiocholine. Both Lineweaver-Burk plots and Dixon plots indicated that the six nondepolarizing neuromuscular blocking agents (NMBAs) at different concentrations induce linear mixed-type inhibition. The apparent inhibition constants (K(i)) of pancuronium (8.72 x 10(-8) M) and vecuronium (3.53 x 10(-7) M) for plasma ChE inhibition were lower than that of neostigmine (7.36 x 10(-7) M), whereas those of the six nondepolarizing NMBAs for erythrocyte AChE were markedly higher than that of neostigmine. CONCLUSIONS: Both plasma ChE and erythrocyte AChE were inhibited by six nondepolarizing NMBAs, and the pattern of inhibition of both enzymes was of mixed type. The inhibitory potencies of pancuronium and vecuronium for plasma ChE were larger than that of neostigmine, whereas those of the six nondepolarizing NMBAs for erythrocyte AChE were markedly lower than that of neostigmine. The rank order of relative potency for plasma ChE was pancuronium > vecuronium > pipecuronium > alcuronium > d-tubocurarine > atracurium. PMID- 14564608 TI - Anesthetic management of a parturient with a diagnosis of chronic renal failure for cesarean section. PMID- 14564609 TI - Thiopental intensifies the euthyroid sick syndrome after cardiopulmonary resuscitation. PMID- 14564610 TI - Dental injury with alveolar bone fracture during mechanical ventilation: a case report. PMID- 14564611 TI - Propofol decreases contractility of isolated blood-perfused left ventricular muscle in the dog. PMID- 14564613 TI - Propofol anesthesia during rhabdomyolysis. PMID- 14564612 TI - Reproducing classical research on the computer is easy. PMID- 14564614 TI - Comparison between in vivo and in vitro pharmacokinetics of succinylcholine in humans. AB - PURPOSE: To compare the in vivo and in vitro pharmacokinetics of succinylcholine (SCh) in humans. METHODS: A bolus of SCh 1 mg.kg(-1) (n = 7) or 2 mg.kg(-1) (n = 11) was given to 18 patients anesthetized with thiopental. Arterial blood samples for determination of in vivo SCh concentrations were collected every 30 s for 5 min. Another 20-ml blood sample was obtained before induction of anesthesia for determination of in vitro SCh. Concentrations of SCh were measured by high performance liquid chromatography. In vivo and in vitro concentrations of SCh vs time data were analyzed by the one-compartment model. RESULTS: The respective in vivo and in vitro pharmacokinetic parameters (SCh 1 mg.kg(-1) vs SCh 2 mg.kg(-1)) were as follows: Plasma clearance was 4.17 +/- 2.37 and 1.85 +/- 0.28 l.min(-1), P < 0.05, vs 2.91 +/- 2.01 and 1.27 +/- 0.43 l.min(-1), P < 0.05. Elimination half-life was 25.4 +/- 10.6 and 47.4 +/- 5.4 s, P < 0.002 vs 26.3 +/- 10.0 and 75.2 +/- 21.8 s, P < 0.00005. CONCLUSION: These results suggest that the rapid disappearance of SCh from the circulation is due to diffusion out of the blood vessels rather than to enzymatic hydrolysis. PMID- 14564615 TI - Anesthesia with sevoflurane, but not isoflurane, prolongs bleeding time in humans. AB - PURPOSE: Halothane has been shown to suppress platelet aggregation in vitro and ex vivo and to prolong bleeding time. In a previous in vitro study, we demonstrated that sevoflurane had a stronger suppressive effect on platelet aggregation than halothane. The present study investigated whether clinical use of sevoflurane affects bleeding time in vivo. METHODS: Thirty-four patients undergoing minor elective surgery were randomly assigned to sevoflurane or isoflurane. Anesthesia was induced with intravenous thiopental and maintained with sevoflurane or isoflurane with nitrous oxide. Bleeding time was measured by the Duke method. An initial (control) measurement was obtained in the operating room before the induction of anesthesia, and a second was obtained 5-10 min after endotracheal intubation but before starting the operation, when the end expiratory concentration of sevoflurane or isoflurane had been stabilized at 1 1.5 times the minimum alveolar concentration (MAC), and the mean arterial pressures were between 80% and 120% of the preanesthetic values. RESULTS: Bleeding time was increased from the preanesthetic value of 2.07 +/- 0.82 min to 2.83 +/- 0.93 min (n = 15) in the sevoflurane group (P < 0.01) but was not significantly altered in the isoflurane group. CONCLUSION: Sevoflurane alters bleeding time in the clinical situation. PMID- 14564616 TI - Quantitative determination of arterial wall mechanics with pulse oximetric finger plethysmography. AB - PURPOSE: The plethysmographic wave of pulse oximetry reflects arterial pulsation at the fingertip. Since arterial pressure and the pulsatile component of the arterial cross section represent the input and output of the fingertip arterial system, respectively, arterial wall mechanics may be delineated from their relationship. We aimed to construct a mathematical model of the fingertip arterial wall from the relationship between the plethysmographic wave and arterial pressure. METHODS: The pulse oximetric plethysmographic signal at the forefinger and pressure at the ipsilateral radial artery were measured. Employing the data, simple mathematical models with one to four mechanical elements were tested in terms of the goodness of fit and the physiological implications. The determined model was applied to the data obtained during administration of vasoactive agents in anesthetized patients. RESULTS: The mathematical model suited for describing arterial wall mechanics was a four-element, two-compartment model. The two compartments represent passive mechanical and active contractile properties, respectively. In the application of the model to the anesthetized patients, the vasoactive agents produced changes in the model parameters that implied mechanical alterations in the arterial wall. CONCLUSION: These findings suggest the benefit of the four-element, two-compartment model in analyzing peripheral vascular wall mechanics influenced by various stimuli in intensive care and anesthesia. PMID- 14564617 TI - Early oral intake after arthroscopic surgery under spinal anesthesia. AB - PURPOSE: We investigated the tolerability of early oral feeding (EOF) and its effects on the recovery of bowel function after spinal anesthesia. METHODS: Thirty-one healthy adult patients undergoing knee arthroscopy or arthroscopic surgeries were randomly assigned to either the EOF group (n = 16) or the nil per os (NPO, n = 15) group. Spinal anesthesia was performed using hyperbaric tetracaine solution in all patients. Patients in the EOF group were allowed free access to solid and liquid food immediately after surgery before analgesia from spinal tetracaine resolved. Oral intake was prohibited for 24 h after completion of surgery in the NPO group. RESULTS: Two patients in each group were mildly nauseated without the need for treatment. While degree of appetite determined by a visual analog scale before the first meal and time to the first gas emission showed no significant differences between groups, the median time to the first defecation in the EOF group (20.6 h) was significantly shorter than that of the NPO group (33.5 h, P = 0.005). No other complications associated with anesthesia, surgery, or EOF were noted. CONCLUSION: Our results suggest that the restriction of EOF after surgery not involving the gastrointestinal tract under spinal anesthesia may not be rational, and that EOF may facilitate recovery of bowel function. PMID- 14564618 TI - Clinical applications of fentanyl pharmacokinetics and pharmacodynamics: roles of fentanyl in anesthesia. PMID- 14564619 TI - Anesthetic management in obese parturients. PMID- 14564620 TI - A case of herpes zoster affecting the glossopharyngeal nerve. PMID- 14564621 TI - Improvement of hypoxia during early surgery for ruptured intracranial aneurysm: a retrospective evaluation. PMID- 14564622 TI - Accidental subdural catheterization suspected on administration of a test dose of lidocaine and successfully managed by a small dose of dibucaine. PMID- 14564623 TI - Implications of the suppression of guanylate cyclase activity by halothane. PMID- 14564625 TI - Orotracheal intubation for a patient with a protruding foreign body in the posterior neck. PMID- 14564626 TI - Folic acid, polymorphism of methyl-group metabolism genes, and DNA methylation in relation to GI carcinogenesis. AB - DNA methylation is the main epigenetic modification after replication in humans. DNA (cytosine-5)-methyltransferase (DNMT) catalyzes the transfer of a methyl group from S-adenosyl-L-methionine (SAM) to C5 of cytosine within CpG dinucleotide sequences in the genomic DNA of higher eukaryotes. There is considerable evidence that aberrant DNA methylation plays an integral role in carcinogenesis. Folic acid or folate is crucial for normal DNA synthesis and can regulate DNA methylation, and through this, it affects cellular SAM levels. Folate deficiency results in DNA hypomethylation. Epidemiological studies have indicated that folic acid protects against gastrointestinal (GI) cancers. Methylene-tetrahydrofolate reductase (MTHFR) and methionine synthase (MS) are the enzymes involved in folate metabolism and are thought to influence DNA methylation. MTHFR is highly polymorphic, and the variant genotypes result in decreased MTHFR enzyme activity and lower plasma folate level. Two common MTHFR polymorphisms, 677CT (or 677TT) and A1298C, and an MS polymorphism, A-->G at 2756, have been identified. Most studies support an inverse association between folate status and the rate of colorectal adenomas and carcinomas. During human GI carcinogenesis, MTHFR is highly polymorphic, and the variant genotypes result in decreased MTHFR enzyme activity and lower plasma folate level, as well as aberrant methylation. PMID- 14564627 TI - Nocturnal gastric acid breakthrough during the administration of rabeprazole and ranitidine in Helicobacter pylori-negative subjects: effects of different regimens. AB - BACKGROUND: Nocturnal gastric acid breakthrough (NAB) is defined as nocturnal intragastric pH less than 4 for more than 1 h during proton pump inhibitor (PPI) administration. A bedtime dose of an H2 receptor antagonist (H2RA) inhibites NAB, but the efficacy of the H2RA decreases with continuous administration. We carried out the present study to investigate the effect of 14-day H2RA administration on NAB. METHODS: Ten male volunteers without Helicobacter pylori infection received four different 14-day regimens of rabeprazole and ranitidine (study a, morning dose of 20 mg rabeprazole; study b, morning dose of 20 mg rabeprazole with a single bedtime dose of 150 mg ranitidine only on the last day; study c, continuous 20 mg morning dose of rabeprazole and 150 mg at bedtime; study d, morning and evening doses of 10 mg rabeprazole). Ambulatory 24-h gastric pH monitoring was conducted on the last day of each regimen. RESULTS: NAB in studies a, b, c, and d was observed in 9, 1, 4, and 4 subjects, respectively, and the longest periods of nocturnal gastric pH at less than 4.0 were 102.5, 14.0, 37.5, and 52.5 min, respectively (study b vs study c, P<0.05). CONCLUSIONS: The continuous inhibitory effect of ranitidine combined with rabeprazole on nocturnal gastric acid secretion declined during 14-day-long administration in H. pylori negative subjects. Split dosing of rabeprazole was more effective than the single morning dose for inhibiting nocturnal gastric acid secretion. PMID- 14564628 TI - Gastric electrical activity in normal neonates during the first year of life: effect of feeding with breast milk and formula. AB - BACKGROUND: To evaluate the effect of feeding with milk on the gastrointestinal tract, we studied gastric electrical activity in 27 healthy fullterm newborns (15 formula-fed newborns and 12 breast-fed newborns) during the first 6 months of life. METHODS: Three-hour electrogastrography (EGG) recordings were performed, using portable equipment, from the third to fifth day after birth until 6 months, at 3-month intervals. The EGG parameters were calculated as raw and integrated data, the latter as AUC of the whole postprandial period. RESULTS: There was a significant difference in the fasting 3-cpm activity between the two groups (repeated measures analysis of variance [ANOVA] P=0.02; multiple comparisons: formula milk at birth vs breast milk at birth P<0.001). In addition, a significant change in the percentage of postprandial bradygastria was found at 6 months, 1 month after weaning (repeated measures ANOVA, P=0.01; multiple comparisons: formula milk at 6 months vs formula milk at 3 months, P=0.03, formula milk at 6 months vs formula milk at birth, P=0.02; breast milk at 6 months vs breast milk at 3 months, P=0.03, breast milk at 6 months vs breast milk at birth P=0.02). CONCLUSIONS: An adult-like gastric 3-cpm activity can be observed in breast-fed newborns in contrast to formula-fed ones, probably as an effect of colostrum. The high bradygastria percentage recorded at 6 months of life might be the result of an increased low-frequency component of the EGG signal because of the transition to a mixed diet. PMID- 14564629 TI - The influence of feeding on gastric acid suppression in Helicobacter pylori positive patients treated with a proton pump inhibitor or an H2-receptor antagonist after bleeding from a gastric ulcer. AB - BACKGROUND: This study investigated the influence of feeding on gastric acid suppression in Helicobacter pylori-positive patients treated with intravenous infusions of proton pump inhibitors (PPIs) or with H2-receptor antagonists (H2 RAs) after bleeding from a gastric ulcer. METHODS: Forty-nine H. pylori-positive patients with bleeding gastric ulcers (44 men and 5 women) were divided into four groups: one group received an H2-RA while fasting, one group received an H2-RA while eating regularly, one group received a PPI while fasting, and one group received a PPI while eating regularly. Intragastric pH was monitored during fasting and nonfasting to calculate the pH 3 and pH 4 holding times and the mean pH. RESULTS: During a 24-h fast, the pH 3 and pH 4 holding times and the mean pH were significantly higher in patients administered omeprazole (PPI; 93.2 +/- 9.2%, 90.6 +/- 11.1%, and 6.9 +/- 0.6, respectively) than in those administered ranitidine (H2-RA; 61.0 +/- 27.5%, 55.8 +/- 29.1%, and 4.8 +/- 1.3, respectively; P<0.001 for all). Results were similar during feeding (PPI meal, 98.9 +/- 2.6%, 98.3 +/- 3.7%, and 6.9 +/- 0.3; H2-RA meal, 59.8 +/- 17.6%, 49.7 +/- 18.0%, and 4.3 +/- 0.7, respectively; P<0.001 for all). In addition, the pH 3 and pH 4 holding times and the mean pH in the H2-RA meal group were not significantly lower than those in the H2-RA group (P=0.999, P=0.865, and P=0.687, respectively). The values in the PPI and PPI meal groups were similar (P=0.872, P=0.777, and P>0.999, respectively). CONCLUSIONS: Gastric acid suppression during the administration of an H2-RA or a PPI soon after the cessation of gastric bleeding was scarcely affected by feeding. It may well be that H. pylori-positive patients with bleeding gastric ulcer can resume a regular diet and return to work soon after bleeding ceases. PMID- 14564630 TI - Real-time energy metabolism of intestine during arterial versus venous occlusion in the rat. AB - BACKGROUND: Although it is less frequently encountered, mesenteric vein occlusion poses as important a problem as mesenteric artery occlusion. The energy metabolism of intestinal tissue during venous occlusion and reperfusion was studied. METHODS: Male Wistar rats were studied in four groups of 17 animals each. Intestinal ischemia was induced by clamping the superior mesenteric artery ([SMA] occlusion [O]) or vein (SMVO) for 30 or 60 min, followed by reperfusion. Magnetic resonance spectroscopy was employed to continuously monitor the energy metabolism. Additionally, intestinal motility was monitored and histological examination was performed on resected specimens. RESULTS: Energy metabolism in SMVO during ischemia was reduced more slowly than in SMAO, but recovery after reperfusion was poorer in SMVO. During ischemia, the contractive response of the intestine lasted longer in SMVO than in SMAO. Histologically, mucosal and subserosal hemorrhage was more severe in SMVO. CONCLUSIONS: In contrast to SMAO, SMVO caused less severe reduction of energy metabolism, at the expense of hemorrhage and tissue damage. PMID- 14564632 TI - Discriminant function for prognostic indexes and probability of death in chronic severe hepatitis B. AB - BACKGROUND: We aimed to determine a discriminant function for prognosis in chronic severe hepatitis B (CSHB), by discriminant analysis of prognostic indexes and probability of death. METHODS: In 205 patients with chronic severe hepatitis B (101 patients in the survival group and 104 patients in the death group), we carried out discriminant analysis of serum total bilirubin, prothrombin activity, white blood cells, creatinine, maximum depth of ascites, hepatic encephalopathy, singultus, and digestive tract hemorrhage. RESULTS: The discriminant function was V=0.00043 x total bilirubin (microM) - 0.025 x prothrombin activity (%) + 0.056 x white blood cells (10(9)/l) + 0.00284 x creatinine (microM) + 0.0014 x maximum depth of ascites (mm) + 0.724 x hepatic encephalopathy score + 0.078 x singultus score + 0.457 x digestive tract hemorrhage score - 2.488. The correctness of the function for predicting death in the death group was 92.3%, and that for predicting survival in the survival group was 96.0%. When the V values were infinity, -4.595, -2.197, -1.386, -0.405, 0.405, 1.386, 2.197, 2.944, 4.595, and infinity, a posterior probabilities of death were 0%, 5%, 10%, 20%, 40%, 60%, 80%, 90%, 95%, 99%, and 100%, respectively. CONCLUSIONS: The discriminant function is an objective, convenient, and practical method to assess the prognosis of chronic severe hepatitis B. PMID- 14564631 TI - Elevated expression of caveolin-1 at protein and mRNA level in human cirrhotic liver: relation with nitric oxide. AB - BACKGROUND: Caveolin, the principal structural protein of caveolae, binds with endothelial nitric oxide synthase (eNOS) leading to enzyme inhibition. This study examined the expression of caveolin and eNOS at the protein and mRNA levels in patients with hepatocellular carcinoma and hepatitis C-related cirrhosis, and in control noncirrhotic liver specimens obtained from patients with metastatic liver carcinoma. METHODS: Anti-eNOS, anti-caveolin-1, and anti-calmodulin antibodies were used for Western blotting. For in situ hybridization (ISH), human eNOS and caveolin-1 peptide nucleic acid probes were used with a catalyzed signal amplification system. RESULTS: Western blotting showed marked overexpression of caveolin-1 protein in cirrhotic liver, while caveolin-1 was almost undetectable in control liver tissue. Endothelial NOS was expressed at a slightly higher level in cirrhotic liver than in control liver tissue. Calmodulin was expressed abundantly in control liver tissue and at a low level in cirrhotic liver tissue. By ISH, eNOS mRNA was localized on portal vein and hepatic lining cells, and caveolin-1 mRNA was almost undetectable in normal liver tissue. In cirrhotic liver tissue, caveolin-1 mRNA was overexpressed on hepatic sinusoidal lining cells, while eNOS mRNA expression was similar to that in normal liver. CONCLUSIONS: Enhanced caveolin-1 expression may be associated with a significant reduction in NO catalytic activity in cirrhosis. PMID- 14564633 TI - Localized giant inflammatory polyposis of the cecum associated with distal ulcerative colitis. AB - This case report describes a 68-year-old man who presented with bronchiolitis obliterans organizing pneumonia (BOOP) and gastric carcinoma. During evaluation, including a colonoscopy, he was found to have distal colitis and a giant polypoid lesion in the cecum that was diagnosed as localized giant inflammatory polyposis (LGIP) by magnifying colonoscopy. The patient was treated over a period of 3 years without surgery, and the LGIP was reduced in size during the follow-up period. Magnifying observation was useful to distinguish LGIP from a neoplastic lesion. PMID- 14564634 TI - Immune thrombocytopenic purpura in patients with ulcerative colitis. AB - Extraintestinal manifestations of ulcerative colitis (UC) are well known, but immunologically mediated hematological diseases are relatively rare. We describe two cases of immune thrombocytopenic purpura (ITP) associated with preexisting UC. Our patients had typical symptoms of UC, and endoscopy showed pancolitis. During treatment with 5-aminosalicylic acid and steroids, severe thrombocytopenia was noted. ITP was diagnosed based on a normal to high number of megakaryocytes in the bone marrow, positive autoantibody to platelet membrane antigen, and absence of splenomegaly. Medical treatment, including increased dosage of steroids, failed to control UC and ITP in both patients. In the first patient, the platelet count recovered after colectomy, while the second patient died of a cerebral hemorrhage. We stress that a diagnosis of ITP should be considered for thrombocytopenia in patients with UC, especially those showing extensive and significant colonic inflammation, and that colectomy of UC might resolve resistant ITP. PMID- 14564635 TI - Crohn's disease complicated by adult-onset Still's disease. AB - A 31-year-old man with Crohn's disease developed arthritis, spiking fever, and skin rash indistinguishable from that of adult-onset Still's disease. He was admitted to our hospital because of a periumbilical intestinal skin fistula. Crohn's disease had been diagnosed in 1991, and had required intestinal resection twice, and schizophrenia had been diagnosed in 1993. He developed polyarthritis and spiking fever, accompanied by a macular skin rash on both forearms. Marked hepatosplenomegaly and bilateral pleural effusion were detected on computed tomography examination. These findings are indistinguishable from those of adult onset Still's disease. Because his mental status had deteriorated following high dose prednisolone on a previous admission, he was treated with an immunosuppressive agent on this occasion, with the treatment being successful. This is the first report of adult-onset Still's disease complicating Crohn's disease. In patients with Crohn's disease, polyarthritis and skin rash can easily be misdiagnosed as enteropathic arthritis with erythema nodosum associated with the Crohn's disease. Although adult-onset Still's disease may not be fatal, early diagnosis is important because it can, in rare cases, result in life-threatening complications. PMID- 14564637 TI - Minute carcinoma of the pancreas presenting as pancreatic pleural effusion. AB - A 70-year-old Japanese man with no history of pancreatitis visited his local practitioner, complaining of dyspnea on effort. Left massive pleural effusion was detected and he was then referred to our hospital. A plain chest film showed marked left pleural effusion. Thoracentesis yielded 2000 ml of bloody fluid with high amylase content (22,665 IU/l). Endoscopic retrograde pancreatography revealed a tapered occlusion of the main pancreatic duct. Pancreatic cancer was suspected, and a distal pancreatectomy and a splenectomy were performed. Histologically, the diagnosis was ductal adenocarcinoma of the pancreas, 5 x 6 mm in size, with regional lymph node metastasis. He has experienced no recurrence of cancer or pleural effusion since the operation. PMID- 14564636 TI - Treatment with STI571, a tyrosine kinase inhibitor, for gastrointestinal stromal tumor with peritoneal dissemination and multiple liver metastases. AB - BACKGROUND: Gastrointestinal stromal tumors (GISTs) are usually refractory to standard chemotherapeutic agents. We successfully treated a patient with a tyrosine kinase inhibitor (STI571) for GIST with peritoneal dissemination and liver metastases. METHODS: In a 32-year-old man presenting with abdominal pain from diffuse peritonitis, a GIST and associated perforated small intestine were resected. Multiple liver metastases were present. After therapies with microwave coagulation, ethanol injection, and local and systemic antineoplastic drugs (fluorouracil, cisplatin, tegafur-uracil, and tegafur) failed, investigational treatment with a tyrosine kinase inhibitor was initiated (STI571, 300 mg, p.o. daily). RESULTS: Anorexia and abdominal fullness resolved within a few days. At 24 days after initiation, positron emission tomography showed a remarkable decrease in the abdominal uptake of [18F] fluorodeoxyglucose. Adverse effects of STI571, including mild alopecia and anemia, were minimal. CONCLUSIONS: The tyrosine kinase inhibitor STI571 may be effective against GISTs. PMID- 14564638 TI - Octreotide in control of multiple liver metastases from gastrinoma. AB - The somatostatin analogue octreotide was effective in controlling systemic effects related to multiple liver metastases from a gastrinoma. A 61-year-old man underwent distal gastrectomy for gastrinoma in the duodenum, because a curative resection was not feasible due to metastases found in paraaortic lymph nodes during operation. Multiple liver metastases, associated with an increase in serum gastrin concentration, were found by magnetic resonance imaging 16 months after the operation. Although chemotherapy with dimethyltrizenoimidazole carboxamide was not effective, subcutaneous administration of octreotide was effective in controlling the growth of the liver metastases and in stabilizing serum gastrin. The patient now receives subcutaneous injections of octreotide, at 200 microg a day, twice a week, as an outpatient. PMID- 14564639 TI - Superficially elevated colonic adenoma changed to undetectable configuration on ordinary endoscopy during treatment with preferential cyclooxygenase-2 inhibitor. PMID- 14564640 TI - Hypertonic saline-epinephrine injection plus endoscopic band ligation therapy for gastric Dieulafoy's lesion. PMID- 14564641 TI - Localized giant inflammatory polyposis (filiform polyposis) with diverticula in ulcerative colitis. PMID- 14564642 TI - Nocturnal gastric acid breakthrough may not be a problem of acid secretion only. PMID- 14564643 TI - Is the incidence of intrahepatic multicentric recurrence of hepatocellular carcinoma more frequent in "the carcinogenic stage" than in liver cirrhosis? PMID- 14564644 TI - Evaluation of an oncology outpatient orientation program: patient satisfaction and outcomes. AB - A person's first visit to an outpatient cancer center is typically marked by considerable stress and negative emotion. Thus, innovative support interventions that aim to ameliorate such stress and emotion are warranted. This study evaluated one such intervention--a new-patient orientation program in a cancer center. The program is designed to: (1) provide patients with information about the center's facilities and procedures, (2) give them an opportunity to ask questions and discuss personal concerns, (3) provide them with detailed information concerning support services available in both the community and the cancer center, and (4) provide them with access to a support care practitioner for follow-up contact. Following the orientation program, 213 participants completed a mailed questionnaire designed to measure their level of satisfaction with various aspects of the orientation program and the effect of the program on their emotional state. The results showed that participants were extremely satisfied with the program, it helped them deal more effectively with their first visit to the center, and it increased their feelings of relaxation and comfort and reduced their feelings of fear and anxiety. These results support the use of informational and support interventions as an effective means of improving cancer care. PMID- 14564645 TI - The role of relaxation time corrections for the evaluation of long and short echo time 1H MR spectra of the hippocampus by NUMARIS and LCModel techniques. AB - 1H MR spectroscopy is routinely used for lateralization of epileptogenic lesions. The present study deals with the role of relaxation time corrections for the quantitative evaluation of long (TE=135 ms) and short echo time (TE=10 ms) 1H MR spectra of the hippocampus using two methods (operator-guided NUMARIS and LCModel programs). Spectra of left and right hippocampi of 14 volunteers and 14 patients with epilepsy were obtained by PRESS (TR/TE=5000/135 ms) and STEAM (TR/TE=5000/10 ms) sequences with a 1.5-T imager. Evaluation was carried out using Siemens NUMARIS software and the results were compared with data from LCModel processing software. No significant differences between the two methods of processing spectra with TE=135 ms were found. The range of relaxation corrections was determined. Metabolite concentrations in hippocampi calculated from spectra with TE=135 ms and 10 ms after application of correction coefficients did not differ in the range of errors and agreed with published data (135 ms/10 ms: NAA=10.2+/ 0.6/10.4+/-1.3 mM, Cho=2.4+/-0.1/2.7+/-0.3 mM, Cr=12.2+/-1.3/11.3+/-1.3 mM). When relaxation time corrections were applied, quantitative results from short and long echo time evaluation with LCModel were in agreement. Signal intensity ratios obtained from long echo time spectra by NUMARIS operator-guided processing also agreed with the LCModel results. PMID- 14564649 TI - Heart rate variability testing, or listening to a conversation in a foreign language. PMID- 14564646 TI - Solubilization of cobalt from ocean nodules at neutral pH-a novel bioprocess. AB - A marine organism ( Bacillus M1) isolated from Indian Ocean manganese nodules was characterized. The organism grew well in artificial seawater medium, at near neutral pH, 30 degrees C and 0.25 M NaCl, and showed MnO(2)-reducing activity. Growing cultures of Bacillus M1 as well as cell-free spent liquor from fully grown cultures were employed to extract metals from the nodules. The spent liquor of cultures of the organism could dissolve around 45% cobalt (Co) at a pH of 8.2 in 2 h. Co recovery by this treatment was comparable to that in acidic leaching with 2.5 M hydrochloric acid solutions, and was independent of pulp density (w/v ratio). The amount of Co dissolved was beyond the thermodynamic solubility limit in aqueous solution at a pH of 8.2. It is inferred that the metabolites present in the spent liquor played a pivotal role in complexing the Fe (III) phase, solubilizing Co in the process. Partial characterization of spent liquor by spot tests, UV visible spectroscopy and FTIR spectroscopy, showed the presence of siderophore-like phenolic compound(s) with an attached carboxyl group that might form soluble organic complexes with Fe (III). PMID- 14564650 TI - Is cerebrovascular autoregulation impaired during neurally-mediated syncope? PMID- 14564651 TI - Determinants of baroreflex sensitivity in health and disease: from correlates to causality. PMID- 14564652 TI - Cardiac neural discharge preceding sudden death: efferent, afferent or both? PMID- 14564653 TI - The effect of pain on changes in heart rate during the Valsalva manoeuvre. AB - The presence of pain may influence autonomic function in patients with painful neurological or cardiovascular disorders. The aim of the present study was to determine whether pain influences cardiac baroreflexes during the Valsalva manoeuvre. Eighteen healthy subjects immersed their hand twice at each temperature in 30 degrees C water and painfully hot (47 degrees C) and cold water (12 degrees C and 7 degrees C) for 2.5 minutes, followed by 5 minute rest periods. During 50% of the immersions, subjects performed the Valsalva manoeuvre (40 mmHg for 30 seconds) starting one minute after their hand entered the water. Pain ratings and heart rate were greater during the 7 degrees C and 47 degrees C immersions than during the 12 degrees C and 30 degrees C immersions. Pain-induced increases in heart rate did not influence peak tachycardia during phase II or III of the Valsalva manoeuvre or peak bradycardia during phase IV, but opposed bradycardia during the post-strain recovery period. Further studies are needed to establish whether pain influences indices of autonomic function during clinical assessment. PMID- 14564654 TI - Cerebral autoregulation during orthostatic stress in healthy controls and in patients with posturally related syncope. AB - Posturally related syncope (PRS) is a common and distressing problem, which frequently occurs in people with no apparent clinical disorder and is ultimately caused by a reduction in blood supply to the brain. The aim of this study was to compare cerebrovascular responses to orthostatic stress in otherwise healthy patients suffering from PRS, and who were shown to have a poor orthostatic tolerance (n=28), with those in healthy control subjects with good orthostatic tolerance (n=11). Responses of heart rate, arterial blood pressure, end tidal carbon dioxide and middle cerebral artery (MCA) blood flow velocity were determined during a progressive orthostatic stress test of combined head-up tilting and lower body suction, which was continued until presyncope. We assessed the efficiency of autoregulation of cerebral blood flow from the relationship between values of MCA velocity and pressure obtained over the expected range for autoregulation (> 55 mmHg). All patients with PRS had a significant correlation between MCA velocity and pressure, but this was seen in only two of the controls. Furthermore, the values of the correlation coefficients were significantly higher in patients than controls, (p<0.001). We interpret these data as indicating that autoregulation in patients with PRS is less effective than in controls and suggest that this provides evidence for a link between abnormalities of regulation of the cerebral circulation and predisposition to syncope. PMID- 14564655 TI - Influence of coronary artery disease risk factors on baroreflex sensitivity in the elderly. AB - In this study, we assessed whether baroreflex sensitivity (BRS) is influenced by risk factors of cardiovascular disease. Subjects of this study were 95 elderly people (40 males and 55 females; mean age +/- SD, 66.6+/-1.6 years) who underwent a medical check-up. BRS was determined as the gain of transfer function in baroreflex arc by spectral analysis of mean blood pressure and R-R interval variabilities in low-frequency band (0.04-0.15 Hz). Gender-related differences in BRS and relationships between BRS and various risk factors of cardiovascular disease were investigated. The value of BRS was significantly higher in males [10.7+/-3.7 (SD) ms/mmHg] than in females [9.0+/-4.0 ms/mmHg, p< 0.05]. However, this gender-related difference disappeared when other variables were taken into account in the multivariate model. Multiple regression analyses showed independent inverse relationships between BRS and heart rate [b=-0.016+/-0.004 (SE) bpm, beta=-0.39], and between BRS and platelet count [b=-0.002+/-0.001 x 103/ micro l, beta=-0.22]. Our results indicated that BRS is inversely related to platelet count in the elderly population. The precise mechanism of this correlation is unknown, but platelet factors released from platelet aggregates can potentially influence vascular function and modify BRS, or there is a common underlying determinant responsible for the covariation. PMID- 14564656 TI - Botulism type B presenting as pure autonomic dysfunction. AB - Botulism nowadays is a rare mostly food-borne disease caused by the toxin of Clostridium botulinum. On the other hand, botulinum toxin blocking cholinergic transmission has become a most powerful treatment option for many focal movement disorders and is increasingly used to treat autonomic disorders [2, 7, 8]. Although muscle weakness is the hallmark of botulism, autonomic dysfunction may be the leading clinical symptom in rare cases and should be included in the differential diagnosis of pure dysautonomia. Here, we describe a patient with autonomic dysfunction as the leading symptom of botulism type B. PMID- 14564657 TI - A neural mechanism for sudden death after myocardial infarction. AB - By monitoring efferent cardiac sympathetic nerve activity (CSNA) directly in a conscious animal we observed, for the first time, that ventricular fibrillation (VF) following myocardial infarction (MI) was preceded by a paroxysm of CSNA which was not baroreflexmodulated. This observation has potential therapeutic implications. PMID- 14564661 TI - Fast adaptive unsharp masking with programmable mediaprocessors. AB - Unsharp masking is a widely used image-enhancement method in medical imaging. Hardware-based solutions can be developed to support high computational demand for unsharp masking, but they suffer from limited flexibility. Software solutions can easily incorporate new features and modify key parameters, such as filtering kernel size, but they have not been able to meet the fast computing requirement. Modern programmable mediaprocessors can meet both fast computing and flexibility requirements, which will benefit medical image computing. In this article, we present fast adaptive unsharp masking on two leading mediaprocessors or high-end digital signal processors, Hitachi/Equator Technologies MAP-CA and Texas Instruments TMS320C64x. For a 2k x 2k 16-bit image, our adaptive unsharp masking with a 201 x 201 boxcar kernel takes 225 ms on a 300-MHz MAP-CA and 74 ms on a 600-MHz TMS320C64x. This fast unsharp masking enables technologists and/or physicians to adjust parameters interactively for optimal quality assurance and image viewing. PMID- 14564662 TI - Cortical responses to cochlear implant stimulation: channel interactions. AB - This study examined the interactions between electrical stimuli presented through two channels of a cochlear implant. Experiments were conducted in anesthetized guinea pigs. Multiunit spike activity recorded from the auditory cortex reflected the cumulative effects of electric field interactions in the cochlea as well as any neural interactions along the ascending auditory pathway. The cochlea was stimulated electrically through a 6-electrode intracochlear array. The stimulus on each channel was a single 80- micro s/phase biphasic pulse. Channel interactions were quantified as changes in the thresholds for elevation of cortical spike rates. Experimental parameters were interchannel temporal offset (0 to +/-2000 micro s), interelectrode cochlear spacing (1.5 or 2.25 mm), electrode configuration (monopolar, bipolar, or tripolar), and relative polarity between channels (same or inverted). In most conditions, presentation of a subthreshold pulse on one channel reduced the threshold for a pulse on a second channel. Threshold shifts were greatest for simultaneous pulses, but appreciable threshold reductions could persist for temporal offsets up to 640 micro s. Channel interactions varied strongly with electrode configuration: threshold shifts increased in magnitude in the order tripolar, bipolar, monopolar. Channel interactions were greater for closer electrode spacing. The results have implications for design of speech processors for cochlear implants. PMID- 14564663 TI - Intradural lumbar disc herniations: the role of MRI in preoperative diagnosis and review of the literature. AB - The goal of this article is to report our experience on intradural lumbar disc herniation, consider the causes of this pathology, and analyze it from clinical, diagnostic, and therapeutic perspectives with a particular emphasis on the role of MRI in preoperative diagnosis. We analyzed nine patients treated surgically for intradural lumbar disc hernia. All of them underwent surgery, and hemilaminectomy was performed. In six cases, the diagnosis of intradural herniation was definitive and, in the three remaining, it was confirmed at surgery. In five cases, CT (with no contrast medium) of the lumbar area revealed disc herniation, but none could it confirm its intradural location. Myelography was performed in two cases but also could not prove intradural extrusion. Magnetic resonance imaging study was used in four cases. In five, the postoperative outcome has been excellent. Patients 6 and 9 recovered anal function postoperatively; patient 6 suffered from occasional and mild micturition urgency. The three patients previously operated (1, 2, 7) showed good outcome. Presently, we believe that radiologic diagnosis of intradural herniation is possible in carefully selected patients, thanks to MRI with gadolinium. PMID- 14564666 TI - Dynamic generation and qualitative analysis of metabolic pathways by a joint database/graph theoretical approach. AB - The dynamic generation and qualitative analysis of metabolic networks relying on continuously growing qualified metabolic data by a joint database/graph theoretical approach is described. The procedure is applied to analyze the connectivity of a metabolic network after enzyme removal and to subsequently perform shortest path analyses. The focus lies on the analysis of the connectivity of the metabolic network depending on model assumptions. Here we analyze the influence of the number of strongly connected components on the assignment of reversibility or irreversibility of the biochemical reactions. PMID- 14564667 TI - Mutations in a novel gene, NHS, cause the pleiotropic effects of Nance-Horan syndrome, including severe congenital cataract, dental anomalies, and mental retardation. AB - Nance-Horan syndrome (NHS) is an X-linked disorder characterized by congenital cataracts, dental anomalies, dysmorphic features, and, in some cases, mental retardation. NHS has been mapped to a 1.3-Mb interval on Xp22.13. We have confirmed the same localization in the original, extended Australian family with NHS and have identified protein-truncating mutations in a novel gene, which we have called "NHS," in five families. The NHS gene encompasses approximately 650 kb of genomic DNA, coding for a 1,630-amino acid putative nuclear protein. NHS orthologs were found in other vertebrates, but no sequence similarity to known genes was identified. The murine developmental expression profile of the NHS gene was studied using in situ hybridization and a mouse line containing a lacZ reporter-gene insertion in the Nhs locus. We found a complex pattern of temporally and spatially regulated expression, which, together with the pleiotropic features of NHS, suggests that this gene has key functions in the regulation of eye, tooth, brain, and craniofacial development. PMID- 14564668 TI - Maspardin is mutated in mast syndrome, a complicated form of hereditary spastic paraplegia associated with dementia. AB - Mast syndrome is an autosomal recessive, complicated form of hereditary spastic paraplegia with dementia that is present at high frequency among the Old Order Amish. Subtle childhood abnormalities may be present, but the main features develop in early adulthood. The disease is slowly progressive, and cerebellar and extrapyramidal signs are also found in patients with advanced disease. Patients have a thin corpus callosum and white-matter abnormalities, as seen on magnetic resonance imaging. Using an extensive Amish pedigree, we have mapped the Mast syndrome locus (SPG21) to a small interval of chromosome 15q22.31 that encompasses just three genes. Sequence analysis of the three transcripts revealed that all 14 affected cases were homozygous for a single base-pair insertion (601insA) in the acid-cluster protein of 33 kDa (ACP33) gene. This frameshift results in the premature termination (fs201-212X213) of the encoded product, which is designated "maspardin" (Mast syndrome, spastic paraplegia, autosomal recessive with dementia), and has been shown elsewhere to localize to intracellular endosomal/trans-Golgi transportation vesicles and may function in protein transport and sorting. PMID- 14564669 TI - Ordered-subsets linkage analysis detects novel Alzheimer disease loci on chromosomes 2q34 and 15q22. AB - Alzheimer disease (AD) is a complex disorder characterized by a wide range, within and between families, of ages at onset of symptoms. Consideration of age at onset as a covariate in genetic-linkage studies may reduce genetic heterogeneity and increase statistical power. Ordered-subsets analysis includes continuous covariates in linkage analysis by rank ordering families by a covariate and summing LOD scores to find a subset giving a significantly increased LOD score relative to the overall sample. We have analyzed data from 336 markers in 437 multiplex (>/=2 sampled individuals with AD) families included in a recent genomic screen for AD loci. To identify genetic heterogeneity by age at onset, families were ordered by increasing and decreasing mean and minimum ages at onset. Chromosomewide significance of increases in the LOD score in subsets relative to the overall sample was assessed by permutation. A statistically significant increase in the nonparametric multipoint LOD score was observed on chromosome 2q34, with a peak LOD score of 3.2 at D2S2944 (P=.008) in 31 families with a minimum age at onset between 50 and 60 years. The LOD score in the chromosome 9p region previously linked to AD increased to 4.6 at D9S741 (P=.01) in 334 families with minimum age at onset between 60 and 75 years. LOD scores were also significantly increased on chromosome 15q22: a peak LOD score of 2.8 (P=.0004) was detected at D15S1507 (60 cM) in 38 families with minimum age at onset >/=79 years, and a peak LOD score of 3.1 (P=.0006) was obtained at D15S153 (62 cM) in 43 families with mean age at onset >80 years. Thirty-one families were contained in both 15q22 subsets, indicating that these results are likely detecting the same locus. There is little overlap in these subsets, underscoring the utility of age at onset as a marker of genetic heterogeneity. These results indicate that linkage to chromosome 9p is strongest in late-onset AD and that regions on chromosome 2q34 and 15q22 are linked to early-onset AD and very-late onset AD, respectively. PMID- 14564670 TI - RP2 and RPGR mutations and clinical correlations in patients with X-linked retinitis pigmentosa. AB - We determined the mutation spectrum of the RP2 and RPGR genes in patients with X linked retinitis pigmentosa (XLRP) and searched for correlations between categories of mutation and severity of disease. We screened 187 unrelated male patients for mutations, including 135 with a prior clinical diagnosis of XLRP, 11 with probable XLRP, 30 isolate cases suspected of having XLRP, and 11 with cone rod degeneration. Mutation screening was performed by single-strand conformation analysis and by sequencing of all RP2 exons and RPGR exons 1-14, ORF15, and 15a. The refractive error, visual acuity, final dark-adapted threshold, visual field area, and 30-Hz cone electroretinogram (ERG) amplitude were measured in each patient. Among the 187 patients, we found 10 mutations in RP2, 2 of which are novel, and 80 mutations in RPGR, 41 of which are novel; 66% of the RPGR mutations were within ORF15. Among the 135 with a prior clinical diagnosis of XLRP, mutations in the RP2 and RPGR genes were found in 9 of 135 (6.7%) and 98 of 135 (72.6%), respectively, for a total of 79% of patients. Patients with RP2 mutations had, on average, lower visual acuity but similar visual field area, final dark-adapted threshold, and 30-Hz ERG amplitude compared with those with RPGR mutations. Among patients with RPGR mutations, those with ORF15 mutations had, on average, a significantly larger visual field area and a borderline larger ERG amplitude than did patients with RPGR mutations in exons 1-14. Among patients with ORF15 mutations, regression analyses showed that the final dark-adapted threshold became lower (i.e., closer to normal) and that the 30-Hz ERG amplitude increased as the length of the wild-type ORF15 amino acid sequence increased. Furthermore, as the length of the abnormal amino acid sequence following ORF15 frameshift mutations increased, the severity of disease increased. PMID- 14564671 TI - Dietary iron regulates hepatic hepcidin 1 and 2 mRNAs in mice. AB - Recently discovered peptide-hepcidin (Hepc) may be a central player in the communication of iron body stores to the intestinal absorptive cells and thus involved in the maintenance of iron homeostasis. The aim of this study was to determine the effects of the level of dietary iron on Hepc gene expression in the liver. OF1 male mice were fed for 3 weeks either control diet (35 mg iron/kg diet), low-iron diet (1 mg iron/kg diet), or high-iron diet (500 mg iron/kg diet), and Hepc 1 and 2 mRNA abundance in the liver was assessed by reverse transcriptase-polymerase chain reaction (RT-PCR). Results clearly showed that Hepc gene expression is upregulated by high dietary iron and downregulated when the dietary iron level is low. Both Hepc 1 and Hepc 2 expression responds coordinately to dietary iron. This work provides additional evidence of the key role of Hepc in the regulation of iron homeostasis. PMID- 14564672 TI - Metabolic fate of extrahepatic arginine in liver after burn injury. AB - Increased nitrogen loss in the form of urea is a hallmark of the metabolic aberrations that occur after burn injury. As the immediate precursor for urea production is arginine, we have conducted an investigation on the metabolic fate of arginine in the liver to shed light on the metabolic characteristics of this increased nitrogen loss. Livers from 25% total surface burn (n = 8) and sham burn rats (n = 8) were perfused in a recycling fashion with a medium containing amino acids and stable isotope labeled l-[(15) N(2)-guanidino, 5,5-(2)H(2)]arginine for 120 minutes. The rates of glucose and urea production and oxygen consumption were measured. The rate of unidirectional arginine transport and the intrahepatic metabolic fate of arginine in relation to urea cycle activity were quantified by tracing the disappearance rate of the arginine tracer from and the appearance rate of [(15)N(2)]urea in the perfusion medium. Perfused livers from burned rats showed higher rates of total urea production (mean +/- SE, 4.471 +/- 0.274 v 3.235 +/- 0.261 mumol. g dry liver(-1). min(-1); P <.01). This was accompanied by increased hepatic arginine transport (1.269 +/- 0.263 v 0.365 +/- 0.021 mumol. g dry liver(-1). min(-1)) and an increased portion of urea production from the transported extrahepatic arginine (12.9% +/- 2.9% v 3.5% +/- 0.4%, P <.05). The disposal of arginine via nonurea pathways was also increased (0.702 +/- 0.185 v 0.257 +/- 0.025 mumol/g dry weight(-1)/min(-1); P <.05). We propose that increased inward transport and utilization of extrahepatic arginine by the liver contributes to the accelerated urea production after burn injury and accounts, in part, for its conditional essentiality in the nutritional support of burn patients. PMID- 14564673 TI - Doxazosin, an alpha-1 antagonist, prevents further progression of the advanced atherosclerotic lesion in hypercholesterolemic hamsters. AB - The aim of this study was to examine the effect of doxazosin (DOX) on the further progression and regression of the advanced atherosclerotic lesion in the hypercholesterolemic hamster. Thirty-six, male F(1)B Golden Syrian hamsters, 10 weeks of age, were divided into 3 groups of 12 and fed a nonpurified hypercholesterolemic diet (HCD) containing 10% coconut oil and 0.1% cholesterol (wt/wt) for 9 months (HCD 9). One group of hamsters was euthanized at 9 months and their aortas were collected, fixed, and stored until analysis. The remaining hamsters were either maintained on the HCD for an additional 6 months (HCD 15) or fed the HCD plus 20 mg/kg/d DOX for the 6 months. At the end of the study (15 months), the DOX-treated hamsters had significantly lower plasma total cholesterol (TC) (-68%), low-density lipoprotein-cholesterol (LDL-C) (-73%), and triglycerides (TG) (-74%) compared with the HCD 15. The lumenal narrowing and intimal thickening atherosclerotic lesions were significantly less in the DOX treated hamsters compared with the HCD 15 (-66% and -70%, respectively). These data suggest that DOX treatment prevents further progression of the advanced atherosclerotic lesion possibly by lowering plasma TC, LDL-C, and TG in hypercholesterolemic hamsters. PMID- 14564674 TI - Platelet count is independently associated with insulin resistance in non-obese Japanese type 2 diabetic patients. AB - The aim of the present study was to investigate the relationship between platelet count and insulin resistance in non-obese Japanese type 2 diabetic patients. A total of 163 non-obese Japanese type 2 diabetic patients (112 men and 51 women, aged 36 to 84 years, body mass index [BMI] 16.2 to 26.9 kg/m(2)) were studied. In conjunction with BMI, glycosylated hemoglobin (HbA(1c)), fasting concentrations of plasma glucose and serum lipids (triglycerides, low-density lipoprotein [LDL] cholesterol, high-density lipoprotein [HDL] cholesterol, and total cholesterol), and hematological parameters (platelets, white blood cell count, red blood cell count, hematocrit, hemoglobin) were measured. LDL cholesterol was calculated using the Friedewald formula. Insulin resistance was estimated by the insulin resistance index of homeostasis model assessment (HOMA-IR). Univariate regression analysis showed that HOMA-IR was positively correlated to BMI (r = 0.465, P <.0001), HbA(1c) (r = 0.423, P <.0001), platelet count (r = 0.310, P <.0001), triglycerides (r = 0.277, P <.0005), white blood cell count (r =.222, P =.005), red blood cell count (r = 0.210, P =.008), hematocrit (r = 0.156, P =.047), total cholesterol (r = 0.178, P =.023), and systolic (r = 0.216, P =.011) and diastolic (r = 0.263, P =.002) blood pressure, and inversely correlated to HDL cholesterol (r = -0.312, P <.0001) level in our diabetic patients. Multiple regression analysis showed that HOMA-IR was independently predicted by BMI (P <.0001, F = 22.45), HbA(1c) (P <.0001, F = 16.15), platelet count (P <.0001, F = 10.75), and serum triglycerides (P <.0001, F = 10.47) levels, which explained 34% of the variability of HOMA-IR in non-obese Japanese type 2 diabetic patients. These results indicate that not only BMI, HbA(1c), and triglycerides levels but also platelet counts are independent predictor of insulin resistance in non-obese Japanese type 2 diabetic patients. PMID- 14564675 TI - Effect of a standardized grape seed extract on low-density lipoprotein susceptibility to oxidation in heavy smokers. AB - The aim of our study was to evaluate the effect of a standardized formulation of a polyphenolic extract of grapes (Leucoselect-Phytosome [LP]) on low-density lipoprotein (LDL) susceptibility to oxidation in a group of heavy smokers. A randomized, double-blind, crossover study was undertaken in 24 healthy male heavy smokers, aged > or = 50 years. Enrolled subjects were given 2 capsules twice daily for 4 weeks (phase 1). Each capsule contained 75 mg of a grape procyanidin extracts and soy-phosphatidlcholine or placebo consisiting of 75 mg lactose and soy-phosphatidlcholine. A wash out period of 3 weeks was then followed by 4 weeks of the opposite treatment (phase 2). Blood samples were taken at baseline and at the end of each phase and assayed for plasma lipids and LDL susceptibility to oxidation. Compliance was good, and no adverse effects were recorded. Subjects did not show significant modification of total cholesterol (TC), triglycerides (TG), high-density lipoprotein-cholesterol (HDL-C) and LDL-C during LP treatment. Among oxidative indices, thiobarbituric acid reactive substances (TBARS) concentration was significantly reduced in subjects taking LP (-14.7% +/- 21.1% v +5.0% +/- 18.1%, P <.01), and the lag phase prolonged (+15.4% +/- 24.4% v -0.1% +/- 16.0%, P <.05) compared with placebo and basal values. The antioxidant potential of grape seed extract polyphenols may prove effective in a model of oxidative stress (smoking); however more investigational data are needed before use in wider clinical settings. PMID- 14564676 TI - Interest of cystatin C in screening diabetic patients for early impairment of renal function. AB - We compared cystatin C, creatinine, and the Cockroft formula for assessment of early renal failure, defined as a (51)Cr-EDTA clearance < 80 mL/min, in 89 diabetic patients with various degrees of renal impairment (glomerular filtration rate [GFR], 11.4 to 196.5 mL/min). The relationships between cystatin C, creatinine, and (51)Cr-EDTA clearance were linearized by plotting the reciprocals of the values, and correlation coefficients were determined. Sensitivity and specificity for the diagnosis of early renal failure were calculated from receiver operating characteristic (ROC) curves. Over the whole population, cystatin C was as well correlated with GFR (r =.74) as was creatinine (r =.67) or the Cockroft formula (r =.88). Moreover, its diagnostic accuracy was comparable to that of the 2 other parameters. Its sensitivity (86.8%) was better than that of creatinine (77.4%) for screening GFR < 80 mL/min, although the Cockroft formula was more sensitive (96.2%). The study of albuminuric diabetics (n = 63) led to similar conclusions, except for a poor sensitivity of cystatin C. In the 36 patients whose plasma creatinine was < 1 mg/dL, 10 (27.7%) had GFR < 80 mL/min. The correlation of creatinine with GFR, its diagnostic accuracy, and sensitivity were significantly lower than those of cystatin C. In this population of patients with normal creatinine levels, the correlation coefficient of cystatin C, its sensitivity, and its diagnostic accuracy were comparable to those of the Cockroft formula. A moderate reduction in GFR may be present in diabetic patients with low creatinine levels. Although Cockroft formula remains the most reliable and the less expensive tool for the evaluation of renal function, cystatin C is a more reliable criterion for screening and assessment than creatinine and represents a useful alternative to the Cockcroft-Gault formula. PMID- 14564677 TI - Effects of dietary restriction on total body, femoral, and vertebral bone in SENCAR, C57BL/6, and DBA/2 mice. AB - Dietary restriction (DR) increases the life span and retards aging, in part, by limiting free radical generation and oxidative damage. DR also reduces body mass, a major determinant of bone mass across the life span. We tested the hypothesis that DR has its most beneficial effects on bone in mouse strains with high free radical generation (sensitive to carcinogenesis [SENCAR] > C57 > DBA) versus the hypothesis that bone mass at weight-bearing sites is determined by body mass in DR and ad libitum (AL)-fed mice. Male mice of each strain were killed at 10 weeks of age (t(0)) or randomized to an AL-fed or 30% DR feeding regimen for 6 months. Food consumption by AL-fed mice was measured daily, and DR mice received 70% of the amount of food consumed by their respective AL-fed mice the previous day. Body fat (%) and bone mineral density (BMD) and content (BMC) were determined by PIXImus densitometry. There were strain-dependent effects on body mass, crown-to rump length, percent body fat, and total body, femoral, and vertebral BMD and BMC under all conditions. SENCAR mice were heavier, longer, had larger bones, and generally exhibited higher total body, femoral, and vertebral BMC and BMD than C57 and DBA mice. DR had beneficial effects on BMD and BMC in the vertebrae of the SENCAR mouse model of high free radical generation and in the obese, diabetes prone C57 mouse model of high end-stage protein glycation. DR DBA and SENCAR mice had lower femoral BMDs and BMCs than their respective AL-fed controls. Regression analysis confirmed linear relationships between total and lean body mass and total body and femoral BMDs and BMCs, suggesting that physiologic adaptation to a lower body mass accounts for the lower femoral bone mineral values observed in DR versus AL-fed mice. Thus, both hypotheses are, at least, partially valid. DR is beneficial in the trabeculae-rich vertebrae of animal models of high oxidant stress, and total/lean body mass determines BMD and BMC in the weight-bearing femur in DR and AL-fed mice. PMID- 14564678 TI - Hypoadiponectinemia is associated with visceral fat accumulation and insulin resistance in Japanese men with type 2 diabetes mellitus. AB - The aim of the present study was to investigate the association of serum adiponectin concentration with regional adiposity and insulin resistance in subjects with type 2 diabetes mellitus. A total of 73 Japanese men with type 2 diabetes (aged 59 +/- 11 years and body mass index [BMI] 23.8 +/- 3.0 kg/m(2), mean +/- SD) were studied. Fasting serum adiponectin and leptin concentrations were determined by radioimmunoassay. Regional adiposity was measured by abdominal computed tomography (CT) at the umbilical level, and insulin resistance was estimated by homeostasis model assessment (HOMA-R). Univariate regression analysis showed that serum adiponectin levels were negatively correlated with subcutaneous and visceral fat areas. With multivariate regression analysis, visceral fat area was a predominant determinant of serum adiponectin levels. In contrast, subcutaneous fat area was strongly associated with serum leptin concentrations. Among subcutaneous and visceral fat areas, BMI, and serum leptin levels, both subcutaneous and visceral fat areas were independently associated with HOMA-R. In another model incorporating serum adiponectin levels, serum adiponectin levels were selected as an independent determinant of HOMA-R instead of visceral fat area. In conclusion, hypoadiponectinemia was associated with visceral fat accumulation rather than subcutaneous fat depot in Japanese men with type 2 diabetes mellitus. Both subcutaneous and visceral fat accumulation contribute to insulin resistance in these subjects, and the contribution of visceral fat may be mediated, in part, by hypoadiponectinemia. PMID- 14564679 TI - Effect of atorvastatin on apolipoprotein B48 metabolism and low-density lipoprotein receptor activity in normolipidemic patients with coronary artery disease. AB - We aimed to examine postprandial dyslipidemia in normolipidemic patients with coronary artery disease (CAD) and the effects of treatment with an hydroxymethyl glutaryl coenzyme A (HMG-CoA) reductase inhibitor (atorvastatin). Subjects with angiographicaly established CAD were randomized to treatment for 12 weeks with 80 mg/d atorvastatin or placebo and the effects on markers of postprandial lipoproteins and low-density lipoprotein (LDL)-receptor binding determined. LDL receptor binding was determined in mononuclear cells, as a surrogate for hepatic activity. Fasting levels of cholesterol (P <.001), LDL-cholesterol (P <.001), apolipoprotein (apo)B(48) (P =.019), remnant-like particle-cholesterol (RLP-C) (P =.032), and total postprandial apoB(48) area under the curve (AUC) (P =.013) significantly decreased with atorvastatin compared with placebo. Atorvastatin also significantly increased LDL-receptor binding activity (P <.001), and this was correlated with changes in fasting apoB(48) (r =.80, P =.01). We report that aberrations in chylomicron metabolism in normolipidemic CAD subjects are correctable with atorvastatin by a mechanism involving increased LDL-receptor activity. This effect may, in part, explain the cardiovascular benefit of statins used in clinical trials of CAD patients with normal lipid levels. PMID- 14564680 TI - Light, but not heavy alcohol drinking, stimulates paraoxonase by upregulating liver mRNA in rats and humans. AB - Paraoxonase 1 (PON) may contribute to the cardioprotective action of high-density lipoprotein (HDL) because it inhibits low-density lipoprotein (LDL) oxidation, a prerequisite for the onset of atherosclerosis. Because light drinking and heavy drinking have diametrically opposite effects on cardioprotection, we have determined the effects of ethanol dosage on rat serum PON activity and its hepatic expression. Furthermore, we have investigated PON activity and polymorphism in human light and heavy drinkers. Our results confirm that HDL-PON inhibited LDL oxidation, destroyed oxidized LDL, and inhibited its uptake by macrophages. Light ethanol feeding caused a 20% to 25% (P <.05) increase in PON activity in both serum and liver and a 59% (P <.001) increase in the level of liver PON mRNA compared with pair-fed control rats. In contrast, heavy ethanol feeding caused a 25% (P <.05) decrease in serum and liver PON activities with a 51% (P <.01) decrease in liver PON mRNA level. Light drinkers had a 395% (P <.001) higher, whereas heavy drinkers had a 45% (P <.001) lower serum PON activity compared with nondrinkers. Significantly, the number of homozygotes versus heterozygotes with respect to high or low activity PON phenotype was similar in all the groups. Therefore, we conclude that light drinking upregulates, whereas heavy drinking downregulates PON activity and its expression, irrespective of its genetic polymorphism. PMID- 14564681 TI - Modification of the relationship between simple anthropometric indices and risk factors by ethnic background. AB - Current targets for body mass index (BMI) and waist circumference (WC) may not be appropriate for those of South Asian origin. The objectives of this study were to determine whether the relationship between BMI and WC with risk factors for cardiovascular disease (CVD) is the same for men and women of South Asian and European descent. Apparently healthy men and women of European (n = 88) and South Asian (n = 93) descent were recruited from 3 hospital communities and assessed for BMI, WC, waist-to-hip ratio (WHR), blood pressure (BP), lipids, insulin, glucose, and CRP. The study cohort was stratified by sex, and regression analyses were performed with individual risk factors as outcomes and ethnicity with either BMI or WC as predictors adjusting for age and height (WC only). BMI and WC were similar between the European and South Asian men and women. South Asian men had significantly higher values for total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), triglycerides (TG), total cholesterol:high-density lipoprotein-cholesterol (HDL-C) and CRP, and significantly lower values of HDL-C. South Asian women had significantly higher values for TG, TC:HDL-C and CRP and significantly lower values of HDL-C, glucose, systolic BP and diastolic BP. In men, ethnicity was an independent predictor for all risk factors except for glucose and insulin, after adjusting for either BMI or WC independent of age and height. For women, ethnicity was an independent predictor for all risk factors except for total cholesterol (WC model only) and insulin (BMI model only), after adjusting for either BMI or WC independent of age and height. The relationship between BMI or WC and risk factors is such that men and women of South Asian descent present with a more adverse risk profile than those of European descent at the same BMI and/or WC. PMID- 14564682 TI - Risk factors for asymptomatic atherosclerosis in Japanese type 2 diabetic patients without diabetic microvascular complications. AB - Atherosclerotic vascular diseases are frequently associated with diabetes mellitus. There has been increasing evidence showing that the atherosclerotic diseases in diabetic patients are distinct from diabetic microvascular complications as to their pathophysiology and epidemiology. However, we have no information on the prevalence of asymptomatic atherosclerosis in diabetic patients before the onset of microvascular diseases. In the present investigation, we aimed to evaluate risk factors for the atherosclerosis in type 2 diabetic patients without the microvascular diseases. For this purpose, we evaluated atherosclerotic change of carotid arteries in 125 Japanese type 2 diabetic patients who had neither atherosclerotic vascular diseases nor diabetic microvascular complications. When atherosclerotic change was defined as the mean intima-media thickness (IMT) of >/= 1.1 mm and/or the presence of plaque lesion, 50% of patients had atherosclerosis of the carotid arteries. Risk factors for the carotid atherosclerosis were age, low-density lipoprotein (LDL)-cholesterol, hypertension, and diabetes treatment. Age and LDL-cholesterol were associated with mean IMT. Age, diabetes treatment, LDL-cholesterol, and hypertension were positively associated with plaque lesion, while high-density lipoprotein (HDL) cholesterol was negatively associated with it. Fasting plasma glucose, glycosylated hemoglobin (HbA(1c)), and known diabetes duration remained unassociated with any parameters of asymptomatic atherosclerosis of the carotid arteries. These results indicate that glycemic control is unrelated with asymptomatic atherosclerosis in type 2 diabetic patients without diabetic microvascular complications. Conventional risk factors and diabetes treatment are independently associated with atherosclerosis of the carotid arteries in these patients. PMID- 14564683 TI - L-thyroxine therapy attenuates the decline in serum triiodothyronine in nonthyroidal illness induced by hysterectomy. AB - It is controversial whether the administration of thyroid hormone to patients with nonthyroidal illness has any beneficial effect. Two groups of patients undergoing abdominal hysterectomy under the same general anesthesia were studied. Group A consisted of 15 women taking chronically l-thyroxine therapy (1.8 mug/kg daily), and group B (control) consisted of 16 apparently healthy euthyroid women taking placebo. Thyroid hormones, cortisol, and interleukin (IL)-6 were measured 1 day before and 1, 2, 3, and 6 days after surgery. Total triiodothyronine (TT(3)) decreased to a significantly greater degree (P <.05) and for a longer period of time in group B than in group A. The significant increase of reverse T(3) (rT(3)) noted early in group B failed to reach the baseline levels until the end of the study, whereas in group A, rT(3) returned to the preoperative values by day 2. Both groups had similar free thyroxine (FT(4)) at baseline. FT(4) increased significantly at day 1 and remained significantly elevated throughout the postoperative period in group B only. Serum TT(4), thyroid-stimulating hormone (TSH), and cortisol did not change significantly in either group. In all patients, IL-6 increased significantly to a peak value at day 1, showing a slow decrease thereafter. A stronger negative correlation was found between T(3) and IL-6 in group B than in group A (r = -.66, P <.0001 v r = -.38, P <.001, respectively) and a strong positive correlation was observed between rT(3) and IL 6 in group B only (r =.57, P <.001). The long-term treatment with T(4) seems to attenuate the decrease of serum T(3), which occurs during the development of nonthyroidal illness postoperatively. The elevation of IL-6 accounted for a greater proportion of the variations of the T(3) and rT(3) in the control group B than in the T(4)-treated group A. PMID- 14564684 TI - Acyl coenzyme A esters differentially activate cardiac and beta-cell adenosine triphosphate-sensitive potassium channels in a side-chain length-specific manner. AB - Recent evidence demonstrates that long-chain acyl coenzyme A esters (CoAs) activate cardiac and beta-cell plasma-membrane (pmK(ATP)) adenosine triphosphate (ATP)-sensitive potassium channels. In this study, we have investigated the differential effects of acyl CoAs of short and medium side-chain length on cardiac and beta-cell pmK(ATP) isoforms. At the single-channel level, the addition of acyl CoAs of differing side-chain length (2 to 16 carbons) to the inside face of membrane patches from ventricular myocytes caused varying increases in pmK(ATP) channel open probability proportional to increases in acyl side-chain length (20 mumol/L acetyl CoA: 310% +/- 90%, 20 mumol/L decanoyl CoA: 570% +/- 150%). A similar dependence of activation on side-chain length was observed in recombinant pmK(ATP) channels (SUR2A/Kir6.2) with full activation of current requiring both the acyl and CoA moieties in the esterified form. We found the recombinant beta-cell K(ATP) channel (SUR1/Kir6.2) to be much less sensitive to medium-chain acyl CoAs (decanoyl CoA: 124% +/- 15% v 231% +/- 25% in SUR2A/Kir6.2), suggesting a role for the cardiac sulfonylurea receptor, SUR2A, in the molecular mechanism of activation by these compounds. We propose that fatty acid metabolism, and the resultant generation of acyl CoAs of varying side-chain length, may be an important regulator of cellular excitability via interactions with the K(ATP) channel. PMID- 14564685 TI - Intra-arterial calcium stimulation test for detection of insulinomas: detection rate, responses of pancreatic peptides, and its relationship to differentiation of tumor cells. AB - The selective intra-arterial calcium stimulation test has greatly facilitated the precise regionalization of insulinomas smaller than 2 cm, which noninvasive techniques (ultrasound [US], computed tomography [CT], magnetic resonance imaging [MRI]) often fail to localize. This study examined not only the role of the test in the localization of insulinomas, but also the responsiveness of 3 beta-cell peptides (insulin, C peptide, and proinsulin) and their relationship to the degree of differentiation of the tumor cells, using percentage decrease of both proinsulin/insulin (P/I) and proinsulin/C peptide (P/C) ratios after stimulation as indices. Ten consecutive surgically proven insulinoma patients each received an injection of calcium into the arteries supplying the pancreas after standard selective angiography and beta-cell peptide levels were measured in samples taken from the right hepatic vein before and 30, 60, 90, 120, and 180 seconds after each injection prior to operation. After surgery, the expressions of the calcium sensing receptor (CaSR) on the resected tumors were assessed by immunohistochemistry. Intra-arterial calcium stimulation with sampling either for insulin or for C peptide correctly predicted the site of insulinoma in 8 of 9 patients or in 7 of 8 patients if the 2 big malignant insulinomas were excluded; thus, the detection rate of this test was 89% and 88%, respectively. Calcium administration stimulated a marked and prompt release of insulin and C peptide simultaneously. Both peaked within 30 to 60 seconds, then declined gradually thereafter, remaining above the baseline at 180 seconds. The magnitude of increase correlated well with the corresponding percentage decrease of P/I and P/C ratios. The response of proinsulin was much less. Immunohistochemistry demonstrated variable membraneous staining for CaSR in normal pancreatic islets and in about 9% of the total normal beta cells, whereas staining in tumor cells was only minimally detectable. We conclude that selective intra-arterial calcium stimulation with hepatic venous sampling either for insulin or for C peptide is a highly sensitive method for the preoperative localization of small insulinomas. Calcium injection stimulates a brisk response of insulin, C peptide, and proinsulin simultaneously and the magnitude of increase of both insulin and C peptide appears to be correlated well with the degree of differentiation of the tumor cells. The exact mechanism by which calcium provokes the release of beta cell peptides is less clear and whether the CaSR is involved in the mechanism of its action requires further study. PMID- 14564686 TI - Effects of estrogen and medroxyprogesterone acetate on subpopulations of triglyceride-rich lipoproteins and high-density lipoproteins. AB - Hormonal replacement therapy (HRT) in postmenopausal women has been shown to increase both triglyceride (TG) and high-density lipoprotein (HDL) cholesterol levels. To better understand the effects of conjugated equine estrogen (CEE) and medroxyprogesterone acetate (MPA), the 2 most commonly prescribed hormones in HRT, on the different subpopulations of TG-rich and HDL lipoproteins, we conducted a placebo-controlled, double-blind, randomized, crossover study consisting of 3 different phases in 14 postmenopausal women. The 3 phases, each 8 week long, included: (1) placebo, (2) CEE 0.625 mg/d, and (3) CEE 0.625 mg/d and MPA 2.5 mg/d. Slight and statistically nonsignificant elevations in TG levels were observed during the CEE treatment. While very-low-density lipoprotein (VLDL) cholesterol levels were not significantly affected by CEE and CEE + MPA, both HRT treatments lowered remnant lipoprotein (RLP) cholesterol (-14% and -37%, respectively). Compared with placebo, CEE caused a significant increase in HDL, HDL(2), apolipoprotein (apo) A-I, LpAI, alpha1, and prealpha1 levels (12%, 27%, 17%, 26%, 60%, and 102%, respectively). The combination therapy blunted the CEE effect on all HDL parameters, resulting in HDL, HDL(2), and LpAI levels being no longer significantly different from placebo. Apo A-I levels and alpha1, and prealpha1 levels were still significantly higher than placebo (+11%, +50%, and +112%, respectively). These results indicate that HRT has beneficial effects on RLP levels and that, while the estrogen component of HRT has a beneficial effect on the HDL subpopulations mostly associated with coronary heart disease (CHD) protection, MPA partially inhibits this effect. PMID- 14564687 TI - Interaction effect of Serine447Stop variant of the lipoprotein lipase gene and C 514T variant of the hepatic lipase gene on serum triglyceride levels in young adults: the Bogalusa Heart Study. AB - The opposing effects of lipoprotein lipase (LPL) Serin447Stop (S447X) polymorphism and hepatic lipase (HL) C-514T polymorphism on serum triglyceride (TG) levels have been known. However, little is known about the interaction effect of these 2 functional gene variants on serum triglyceride levels. This aspect was examined in a community-based sample of 902 whites and 389 blacks aged 18 to 41 years, using a repeated measures analysis in a mixed model. The frequency of the LPL X447 allele was higher in whites than blacks (16% v 11%, P <.05); whereas the frequency of HL T-514 allele was higher in blacks than whites (77% v 40%, P <.001). The combined genotype distribution was also different between whites and blacks (P <.001). Although the frequency of carriers of both variants was similar in whites and blacks (7% v 8%), more whites carried the LPL X447 allele only (9% v 3%), and more blacks carried the HL T-514 allele only (70% v 33%). Mean levels of TG adjusted for age, sex, and body mass index (BMI) in carriers versus noncarriers of the LPL X447 allele were lower by 13.5% (P <.0001) in whites, 15.8% (P <.01) in blacks and 16.0% (P <.0001) in the total sample. No such phenotypic effect was noted with respect to HL T-514 allele either in blacks or whites, although the mean level in carriers was marginally (P =.08) higher in the total sample. The interaction effect of LPL and HL variants on TG levels was significant in the total sample (P =.016) and marginal in whites (P =.079). In the total sample, the decrease of TG in carriers versus noncarriers of the LPL X447 was 1.8-fold greater in carriers versus noncarriers of the HL T-514 allele (13.6 mg/dL v 7.4 mg/dL, P =.016). Whites tended to show a similar trend (16.8 mg/dL v 6.1 mg/dL, P =.079). Blacks also showed a similar, but nonsignificant, trend (10.4 mg/dL v 8.6 mg/dL, P =.45). These results by showing modulation of association between S447X variant of the LPL gene and serum TG by C-514T variant of the HL gene underscore the importance of gene-gene interactions in the assessment of genetic effects on complex traits. PMID- 14564688 TI - Lipodystrophy in human immunodeficiency virus patients impairs insulin action and induces defects in beta-cell function. AB - The pathophysiology of insulin resistance in human immunodeficiency virus (HIV) associated lipodystrophy syndrome (HALS) is not fully clarified. We investigated 18 men with HALS and 18 HIV-positive males without lipodystrophy (control subjects). Duration and modality of antiretroviral therapy were similar between study groups. A hyperinsulinemic euglycemic clamp showed an impaired glucose disposal rate (GDR) in HALS patients (5.6 v 8.3 mg glucose/min. kg(FFM), P =.0006). As demonstrated by indirect calorimetry, HALS patients showed an impaired nonoxidative glucose metabolism (NOGM, 2.2 v 4.2, P =.006), whereas levels of basal and insulin-stimulated oxidative glucose metabolism (OGM) (2.4 v 2.3, P =.55, and 3.3 v 4.0, P =.064, respectively) were not significantly different between groups. Despite comparable total fat masses, dual energy x-ray absorptiometry (DEXA) scans showed that the percentage of limb fat (ie, peripheral-fat-mass/[peripheral-fat-mass + trunk-fat-mass]. 100%) was reduced in HALS patients (36% v 46%, P =.0002). Multiple linear regression analysis indicated that percentage of limb fat explained 53% of the variability of GDR and 45% of the variability of NOGM in HALS patients. In HALS patients, leg fat mass correlated positively with NOGM (r =.51, P <.05), whereas abdominal fat mass and NOGM did not correlate (P =.91). Analyzing the relationship between first phase insulin secretion and insulin sensitivity, 6 HALS patients compared with none of the control subjects exhibited impaired insulin secretion (P <.05). Our data suggest that fat redistribution independently of antiretroviral therapy is highly related to insulin resistance in HALS patients. Furthermore, in HALS patients, impaired glucose metabolism most likely relates to decreased NOGM and to defects in beta-cell function. PMID- 14564689 TI - Apolipoprotein CIII deficiency prevents the development of hypertriglyceridemia in streptozotocin-induced diabetic mice. AB - To explore the role of apolipoprotein (apo) CIII in the development of hypertriglyceridemia associated with diabetes mellitus, we examined triglyceride (TG) kinetics in apo CIII - deficient mice (apo CIII - null) and wild-type (WT) (C57BL/6J) mice with diabetes induced by the injection of streptozotocin (STZ). Plasma TG levels increased significantly in WT mice after diabetes was induced (102 +/- 29 v 65 +/- 33 mg/dL, P <.01). Apo CIII-null mice had a significantly lower TG level (35 +/- 9 mg/dL) that remained unchanged even when diabetes was induced (35 +/- 8 mg/dL). The TG secretion rate (TGSR) measured by the Triton WR1339 method tended to decrease in diabetic WT, indicating that catabolism of TG was impaired. Apo CIII-null mice showed 2-fold higher TG production than WT mice, indicating markedly faster clearance of TG. The high TGSR was halved when diabetes was induced in apo CIII-null mice, and the fractional catabolic rate (FCR) of TG was also halved, although it was still significantly higher than in WT mice. Lipoprotein lipase (LPL) activity in postheparin plasma was not significantly altered in WT or apo CIII-null mice regardless of the presence or absence of diabetes. [(3)H] very-low-density lipoprotein (VLDL)-TG from WT or apo CIII-null mice showed similar clearance by WT recipients, and this was also observed when VLDL was obtained from diabetic counterparts. In contrast, VLDL-TG was cleared faster by apo CIII-null recipients compared with WT recipients, regardless of the VLDL donors. These results suggest that apo CIII deficiency prevents the development of hypertriglyceridemia associated with diabetes by stimulating TG removal, possibly by promoting the interaction of VLDL with the TG removal system. PMID- 14564690 TI - Acylation-stimulating protein precursor proteins in adipose tissue in human obesity. AB - Recent reports have suggested a link between acylation-stimulating protein (ASP) and complement C3 with obesity, insulin resistance, coronary artery disease, and hyperlipidemia. Our aim was to examine the mRNA expression of C3 and other factors related to ASP production (such as factor B and adipsin) in adipose tissue. The influence of gender and obesity was examined in subcutaneous (SC) and omental (OM) tissues from 16 males and 16 females with body mass index (BMI) from 20 to 54 kg/m(2). The results demonstrate that factor B mRNA expression is higher in males than females in both SC and OM tissues. In female SC tissue, C3 and adipsin mRNA decrease with increasing BMI (r = 0.557, P =.025 and r = 0.717 P =.002, respectively), with no change in factor B. By contrast, in males there was a pronounced increase in C3, adipsin, and factor B in OM tissue with increasing BMI (r = 0.759 P =.001, r = 0.650 P =.006, and r = 0.568 P =.022, respectively). Of note, however, in both men and women there was a marked increase in the OM/SC ratio of C3 and adipsin with increasing BMI. These results suggest that in female SC adipose tissue, there is downregulation of factors related to ASP production in obesity, perhaps to limit further expansion of adipose tissue. In males, there is increased expression in OM tissue. In addition, relative OM/SC expression increases with obesity and these changes may contribute to the development of visceral adipose tissue. PMID- 14564691 TI - Palmitate potentiation of glucose-induced insulin release: a study using 2 bromopalmitate. AB - The mechanisms whereby fatty acids (FA) potentiate glucose-induced insulin secretion from the pancreatic beta cell are incompletely understood. In this study, the effects of palmitate on insulin secretion were investigated in isolated rat islets. Palmitate did not initiate insulin secretion at nonstimulatory glucose concentrations, but markedly stimulated insulin release at concentrations of glucose > or = 5.6 mmol/L. At concentrations of palmitate > or =0.5 mmol/L, the important determinant of the potency of the FA was its unbound concentration. At total concentrations < or = 0.5 mmol/L, both the total and unbound concentrations appeared important. Surprisingly, 2-bromopalmitate did not affect palmitate oxidation, but significantly diminished palmitate esterification into cellular lipids. Neither methyl palmitate, which is not activated into a long-chain acyl-CoA ester, nor 2-bromopalmitate affected glucose-stimulated insulin release. Further, 2-bromopalmitate partly inhibited the potentiating effect of palmitate. These results support the concept that FA potentiation of insulin release is mediated by FA-derived signals generated in the esterification pathway. PMID- 14564694 TI - Purity of radiochemicals. PMID- 14564695 TI - Phospholipids of rat tissues after feeding pure phosphatidyl ethanolamine and lecithin. AB - Pure phosphatidyl ethanolamine and lecithin from egg yolks were fed to rats in saline or in olive oil and the changes in individual phospholipids in the intestinal wall, liver, and plasma of the animals were studied. Ingestion of olive oil alone produced increased levels of all phospholipid fractions in each of the three tissues. Feeding phosphatidyl ethanolamine in saline resulted in slightly increased plasma phospholipids, but levels of liver total phospholipids were greatly reduced; when phosphatidyl ethanolamine was fed with olive oil, liver phospholipids were again reduced but this reduction was confined to the phosphatidyl ethanolamine and phosphatidic acid fractions. Feeding lecithin alone did not produce significant changes in levels of plasma or tissue phospholipids. The results suggest that liver phospholipid synthesis is depressed by feeding phosphatidyl ethanolamine; in the presence of olive oil, hepatic synthesis of phosphatidyl ethanolamine seems to be more selectively inhibited. PMID- 14564698 TI - Conversion of fatty aldehyde dimethyl acetals to the corresponding alk-1-enyl methyl ethers (substituted vinyl ethers) during gas-liquid chromatography. AB - The behavior of palmitaldehyde and linolealdehyde and of their dimethyl acetals during gas-liquid chromatography on beta-cyclodextrin acetate (beta-CDX acetate) and ethylene glycol succinate polyester-phosphoric acid (EGSP) columns was studied. The aldehydes were well separated from their dimethyl acetals on the beta-CDX acetate column. However, on the EGSP column the retention times of palmitaldehyde and its dimethyl acetal were identical; a mixture of linolealdehyde and its dimethyl acetal gave a split peak. The aldehydes were recovered unchanged in 80-85% yield by preparative GLC from both columns, but the dimethyl acetals were quantitatively converted to the corresponding alk-1-enyl methyl ethers. The structure of these compounds was elucidated by infrared spectroscopy, mass spectrometry, and chemical means. Upon hydrolysis at low temperatures with 100% H(2)SO(4) they yielded the corresponding aldehydes, which were identified as 2,4-dinitrophenylhydrazones. PMID- 14564699 TI - Turnover of cholesteryl esters of plasma lipoproteins in the rat. AB - Turnover of individual classes of cholesteryl esters (classified on the basis of the degree of unsaturation of the fatty acid moiety) in rat plasma lipoproteins and liver was studied after the administration of mevalonic acid-5-(3)H and mevalonic acid-2-(14)C. The relative turnover rate was greatest in the d < 1.019 lipoproteins, with monoenes > saturated = dienes > tetraenes. In the d > 1.063 lipoproteins, all cholesteryl esters had slower turnover rates, but tetraenes = pentaenes > dienes > monoenes = saturated. Comparisons of specific activities of individual cholesteryl ester classes of liver subcellular fractions and lipoproteins suggest that the d < 1.019 lipoprotein cholesteryl esters are synthesized from newly synthesized cholesterol in the liver and are rapidly released into this lipoprotein. Tetraenoic cholesteryl esters, however, may originate from esterification of free cholesterol in plasma. Tetraenoic esters are formed from cholesterol in plasma during incubation or ultracentrifugation unless a thiol-reacting or alkylating agent is added. Failure to add such a reagent to plasma results in erroneous specific activities. In the adrenal, relative rates of synthesis of cholesteryl esters are monoenes = dienes > tetraenes > trienes = pentaenes > saturated. It is concluded that cholesteryl ester turnover in the rat, as opposed to man, is determined not only by the particular lipoprotein class but also by the fatty acid moiety of the ester. PMID- 14564700 TI - Conversion of cholesterol injected into man to cholestanol via a 3-ketonic intermediate. AB - Cholesterol-3-(3)H,4-(14)C was injected intravenously in man and its transformation to cholestanol was studied. From the (3)H: (14)C ratios in cholestanol isolated from blood, evidence for the participation of a ketonic intermediate in the conversion was obtained. In a second subject given cholestanol-3-(3)H,4-(14)C the (3)H: (14)C ratios in blood sterols remained unchanged for as long as 1 wk after the injection, which showed that cholestanol did not lose tritium by interconversion with cholestanone. PMID- 14564701 TI - Hypolipidemic effects of orally administered dextran and cellulose anion exchangers in cockerels and dogs. AB - Various cellulose and dextran anion exchangers bind bile salts in vitro under conditions of pH and ionic strength resembling those in the lumen of the small intestine. Of these substances, diethylaminoethyl (DEAE) cellulose, guanidoethyl cellulose, and DEAE Sephadex reduced hypercholesterolemia when added to the diet of cholesterol-fed cockerels. In addition, DEAE Sephadex reduced serum sterols in normocholesterolemic cockerels and dogs, lowered serum phospholipids and triglycerides in cholesterol-fed hypercholesterolemic cockerels and in normocholesterolemic dogs, and increased fecal excretion of bile acids in hypercholesterolemic cockerels. The data indicate that these insoluble cationic polymers exert their hypocholesterolemic effects by interrupting the enterohepatic circulation of bile acids. PMID- 14564702 TI - Pyridine nucleotide synthesis by rat adipose tissue in vitro. AB - The synthesis of NAD and NADP by rat adipose tissue was measured in vitro. Nicotinamide-7-(14)C and NaH(2)(32)PO(4) were incorporated together into NAD with a (32)P/(14)C ratio of 1.82 and nicotinic-7-(14)C acid and NaH(2)(32)PO(4) with a ratio of 1.94. Nicotinic acid stimulated, by 90%, lipogenesis from glucose-U (14)C by rat adipose tissue in vitro. Glucose plus insulin and refeeding for 48 hr after a 48 hr fast markedly increased the incorporation of nicotinic-7-(14)C into NAD in rat epididymal fat pads in vitro, but neither fructose, L-glutamine, nor insulin alone increased the synthesis of NAD in this tissue. Glucose-1-(14)C, ribose-1-(14)C, and to a greater extent glucose-6-(14)C are incorporated into the NAD of rat adipose tissue. Fasting followed by refeeding sharply increased the radioactivity of NAD-(14)C formed from glucose-1-(14)C and glucose-6-(14)C but not from ribose-1-(14)C. Increasing the ribose concentration from 2 mM to 10 mM increased its incorporation into adipose tissue NAD twofold. The nicotinic-7 (14)C acid incorporation into NAD increased over the 1st hr of incubation and remained constant for the next 3 hr. The concentration of NAD in the fat pads showed a similar response to the time of incubation. NADP concentrations increased over the entire 4 hr incubation period as did the incorporation of nicotinic-7-(14)C acid into NADP. The results of this study suggest that NAD is synthesized de novo by rat adipose tissue in vitro and that this synthesis is increased by factors which stimulate lipogenesis. PMID- 14564703 TI - Isolation and analysis of free fatty aldehydes from rat, dog, and bovine heart muscle. AB - Procedures are described for the isolation of free fatty aldehydes from total lipid extracts of rat, dog, and bovine heart muscle. These aldehydes did not arise from hydrolysis of the naturally occurring alkenyl ethers in either the extraction or isolation procedures. The free fatty aldehydes were present in heart muscle in amounts between 0.27 and 0.56 micromole per 100 mg of lipid. Hexadecanal and octadecanal are the principal free fatty aldehydes present, except in dog heart muscle where an unidentified, unusual aldehyde constitutes 20% of the mixture. PMID- 14564704 TI - Post-heparin serum lecithinase in man and its positional specificity. AB - Lecithinase activity in post-heparin serum has been demonstrated. Phosphatidyl choline (PC) can be degraded to lysophosphatidyl choline and fatty acids at a rate of more than 1 micromole/hr per ml of serum in an incubation system containing PC, 0.1 M glycine-NaOH buffer (pH 9.6), and deoxycholate. This activity cannot be found in serum obtained prior to the injection of heparin. Post-heparin serum lecithinase can be distinguished from the heat-stable pancreatic lecithinase by the markedly different effects of heat, paraoxon, and EDTA, and from serum lecithin:cholesterol acyltransferase by the differential effect of p-hydroxymercuribenzoate. In contrast to the acyltransferase and to pancreatic lecithinase, which are active at the beta (C-2) position of lecithin, post-heparin serum lecithinase is active at alpha' (C-1) position. PMID- 14564705 TI - Rapid quantitative procedure for removing cholesterol from butter fat. AB - Cholesterol is rapidly and quantitatively removed from butter fat by passage of the fat in hexane or benzene solution over a column of Celite impregnated with an aqueous solution of digitonin. PMID- 14564706 TI - Efficient extraction of bile acid conjugates with tetraheptylammonium chloride, a liquid ion exchanger. AB - Ethyl acetate or chloroform solutions of tetraheptylammonium chloride, an oil soluble quaternary amine, quantitatively extract polar, anionic lipids such as steroid or bile salt conjugates from aqueous solution by a process of anion exchange. PMID- 14564707 TI - New type of needle for obtaining large samples of human adipose tissue. AB - A new type of needle for obtaining human fatty tissue samples is described. With this instrument 350-600 mg of tissue may be obtained less traumatically than with previously described instruments. PMID- 14564708 TI - Calibration of flow-through detectors of 14C in gas-liquid chromatographic effluents. AB - A simple procedure for calibrating flow-through detectors of radioactivity in the effluent of a gas chromatography column is described. PMID- 14564709 TI - Synthesis of glycerol 1,3-dihexadecyl ether. AB - The synthesis of 1,3-dihexadecyloxy-2-propanol (glycerol 1,3-dihexadecyl ether) is reported. The method is applicable to the preparation of other 1,3 disubstituted glycerols where the substituents are not affected by acid or by catalytic hydrogenolysis conditions. PMID- 14564710 TI - An improved preparation of bovine brain proteolipid. AB - An improved preparation of proteolipid from bovine brain white matter is described. The product obtained by repeated acetone precipitation is completely soluble in chloroform-methanol and has a fairly constant composition: 35% protein, 40% galactocerebroside, and about 25% phospholipid. PMID- 14564711 TI - The citrate cleavage pathway and lipogenesis in rat adipose tissue: replenishment of oxaloacetate. AB - Fatty acid synthesis via the citrate cleavage pathway requires the continual replenishment of oxaloacetate within the mitochondria, probably by carboxylation of pyruvate. Malic enzyme, although present in adipose tissue, is completely localized in the cytoplasm and has insufficient activity to support lipogenesis. Pyruvate carboxylase was found to be active in both the mitochondria and cytoplasm of epididymal adipose tissue cells; it was dependent on both ATP and biotin. Alteractions in dietary conditions induced no significant changes in mitochondrial pyruvate carboxylase activity, but the soluble activity was depressed in fat-fed animals. The possible importance of the soluble activity in lipogenesis lies in its participation in a soluble malate transhydrogenation cycle with NAD malate dehydrogenase and malic enzyme, whereby a continual supply of NADPH is produced. Consequently, the pyruvate carboxylase in adipose tissue both generates mitochondrial oxaloacetate for the citrate cleavage pathway and supplies soluble NADPH for the conversion of acetyl-CoA to fatty acid. PMID- 14564712 TI - Quantitative and compositional changes in monogalactosyl and digalactosyl diglycerides during light-induced formation of chloroplasts in Euglena gracilis. AB - The formation of chloroplasts in dark-grown cells of Euglena gracilis was induced by exposing the cells to constant illumination. Following a lag, the cells accumulated chlorophyll and galactosyl diglycerides simultaneously at almost linear rates. The monogalactosyl diglyceride content rose from approximately 2 micromoles in 100 mg of dark-grown cells to 27 micromoles in fully green cells; the digalactosyl diglyceride content increased from 1 micromole to 11 micromoles. The digalacto compounds increased more rapidly than the monogalacto compounds at first, but their rate of accumulation began to diminish long before greening of the cell was complete. The sole exception was the digalactosyl diglyceride fraction that contained hexadecadienoic (16:2) fatty acid. This fraction increased continuously during greening. As accumulation of the digalacto compounds diminished, that of the monogalacto compounds increased. Towards the end of greening, the major fatty acids were 16:2, 16:3, 16:4, 18:2, and 18:3 in the monogalacto and 16:2 in the digalacto compounds. The results of this study suggest that monogalactosyl and digalactosyl diglycerides that contain particular fatty acid components have a function in the assembly of chloroplasts. PMID- 14564713 TI - Lipogenesis from amino acids in perfused isolated dog skin. AB - Lipogenesis from amino acids has been studied in isolated perfused dog skin. Uniformly labeled alanine-(14)C, glycine-(14)C, isoleucine-(14)C, leucine-(14)C, phenylalanine-(14)C, and valine-(14)C are all incorporated into the cutaneous lipids, with significant incorporation into most of the isolated lipid fractions. Efficiency of lipogenesis has been expressed relative to the extent of incorporation of acetate under the same experimental conditions. This efficiency was highest for the three branched-chain amino acids. The accuracy, uses, and limitations of the perfusion technique for the study of cutaneous lipogenesis have been evaluated. PMID- 14564714 TI - Metabolism of chylomicrons by the isolated rat liver. AB - Isolated livers perfused with washed corn oil chylomicrons labeled in vivo with palmitic acid-1-(14)C removed a large proportion of the chylomicrons. Slices from these livers oxidized chylomicron fatty acid esters to both carbon dioxide and acetoacetate. The liver slices also generated free fatty acids from chylomicron lipids and converted chylomicron triglycerides to phospholipids. Similar activities were observed in rat liver slices prepared shortly after the intravenous administration of chylomicrons to intact rats. The observed chylomicron uptake and lipid conversions were similar in livers from both fed and fasted rats. Fasting increased the oxidation of chylomicron fatty acid esters by livers labeled in vivo and by perfusion. In livers removed from intact rats given labeled chylomicrons, the triglyceride-(14)C to phospholipid-(14)C ratio was high, a finding unexpected if the liver had acquired this (14)C by removal of circulating fatty acids formed by extrahepatic lipolysis. These results demonstrate the ability of the liver to remove and utilize chylomicrons directly and suggest that direct removal accounts for a significant portion of the chylomicron fatty acids utilized by the liver of intact rats. PMID- 14564715 TI - Effect of cholesterol feeding and fasting on sterol synthesis in seventeen tissues of the rat. AB - Rates of sterol synthesis were measured in 17 tissues of the rat, and the responsiveness of these rates to cholesterol feeding and to fasting was determined. The liver and gastrointestinal tract together account for 90% of synthetic activity of the whole body. After the rats had been fed cholesterol or fasted, liver synthesis was markedly decreased, whereas synthetic rates in all other organs tested were essentially unaffected (this conclusion applies to synthesis of cholesterol and of five other digitonin-precipitable tissue sterols). Consequently, the highest rate of cholesterogenesis in the cholesterol fed or fasted rat is found in the gastrointestinal tract. PMID- 14564716 TI - Brain ceramide hexosides in Tay-Sachs disease and generalized gangliosidosis (GM1 gangliosidosis). AB - The carbohydrate composition was determined for ceramide hexosides isolated from brains of patients with Tay-Sachs disease and generalized gangliosidosis (hereby named GM1-gangliosidosis). Gray matter of patients with each disease showed a characteristic abnormal ceramide hexoside pattern. In Tay-Sachs gray matter, ceramide trihexoside is the major component, whereas ceramide tetrahexoside is barely detectable. In GM1-gangliosidosis, ceramide tetrahexoside is the major ceramide hexoside, while ceramide trihexoside is present only in small amount. These two major components have been characterized as the asialo derivatives of, respectively, the "Tay-Sachs ganglioside" (GM2-ganglioside) and the normal major monosialoganglioside (GM1-ganglioside). In both diseases, more than half the ceramide monohexoside of gray matter was glucocerebroside. Gray matter ceramide dihexoside, present in both diseases at higher than normal levels, was mostly ceramide lactoside, with possibly a small amount of ceramide digalactoside. Sulfatide contained only galactose. The abnormal ceramide hexoside pattern is limited to gray matter: white matter showed normal ceramide hexosides, i.e. a preponderance of monohexosides and sulfatide, with no detectable glucocerebroside. PMID- 14564717 TI - Endogenous triglyceride turnover in liver and plasma of the dog. AB - Radioactive glycerol and S(f) > 20 lipoproteins labeled with it were used to study turnover of plasma S(f) > 20 and hepatic triglyceride in anesthetized dogs. From specific activity-time curves of these lipids after an injection of labeled material, a tentative and incomplete model for the kinetics of endogenous hepatic and plasma triglyceride was defined and partially validated. Pool sizes and turnover rates of triglyceride in liver and S(f) > 20 lipoproteins of plasma were then calculated in seven dogs. Hepatic triglyceride was composed of two compartments: 60% metabolically inert and 40% metabolically active. Although communication between these hepatic compartments surely occurred during the time course of these studies, it was not sufficient to be detected by our present methods. The metabolically active compartment turned over as a single pool but with two destinations: a quite variable proportion (an average of 61%) was secreted into plasma as S(f) > 20 triglyceride, and an average of 39% was presumably hydrolyzed within the liver. The fractional turnover rate of plasma S(f) > 20 triglyceride was 2-3 times that of hepatic triglyceride. This finding, and the parallel decline of specific activities of plasma S(f) > 20 and liver triglyceride after injection of labeled glycerol, confirm the rate-determining role of hepatic triglyceride. In this respect the dog differs importantly from man. Though turnover rates of plasma S(f) > 20 triglyceride fell in the same range in men and dogs, the relationship of turnover rate to plasma concentration of this lipid differed greatly between them. The model for the dog does resemble that previously reported for man, however, in the lack of major recycling of intact plasma triglyceride between the liver and plasma. Lack of such recycling, however, does not exclude return of plasma triglyceride into a hepatic triglyceride sink. The amount of such unidirectional uptake, if any, could not be determined by these techniques. PMID- 14564719 TI - The uropygiols: identification of the unsaponifiable constituent of a diester wax from chicken preen glands. AB - The chief lipid fraction in the uropygial gland excretion of the domestic hen is a diester wax. The saponifiable fraction of this wax consists of saturated normal C(10)-C(20) fatty acids. The unsaponifiable fraction consists of a series of three homologous compounds, which have been named the uropygiols and identified by mass spectrometry, gas-liquid chromatography, and periodate cleavage as 2,3-n alkanediols containing 22-24 carbon atoms. The native diols were shown to consist of about equal amounts of the threo and erythro isomers. Records of analyses of the natural products as well as related synthetic compounds are shown. PMID- 14564718 TI - Sex differences in plasma cholesterol-esterifying activity in rats. AB - Esterification of free cholesterol was investigated after incubation at 37 degrees C of plasma from immature and adult rats of both sexes kept on stock, fat free, or cholesterol-supplemented diets. According to measurements of the decrease in free cholesterol, plasma from the fat-deficient rats showed the highest cholesterol-esterifying activity. Esterification was higher in the mature female rats than in the mature males on stock or cholesterol-containing diets, although no sex differences were observed in the sexually immature young or in the fat-free animals. There were no sex differences in the fatty acid composition of the plasma sterol esters, phospholipids, and triglycerides in the immature animals, but arachidonic acid increased at the expense of linoleic acid in the sterol ester fraction in the adult female (not, however, in the adult male). In the phospholipid fraction the higher ratio of palmitic to stearic acids in the male was confirmed. There was an increase in linoleic acid in all three plasma lipid fractions of the mature male after cholesterol feeding. It is suggested that cholesterol may inhibit the conversion of linoleate to arachidonate. During the incubation of plasma, there was little change in the distribution of fatty acids except for a decrease in palmitoleate, and increases in C(20) tri- and tetraenoic acids, in the sterol esters of mature female rats on the stock ration and the fat-free diet. These C(20) acids decreased concomitantly in the phospholipid fraction, as the transesterification reaction mechanism proposed by earlier workers would predict. PMID- 14564720 TI - Serum lipoproteinemia in pregnant and lactating rats. AB - Serum lipoproteins of pregnant and puerperal rats were studied by preparative and analytical ultracentrifugation. The concentration for the fraction with density less than 1.019 was markedly elevated in rats during the 3rd wk of gestation and in lactating rats. This fraction showed similar triglyceride fatty acid composition and immunoelectrophoretic behavior whether it was derived from pregnant or nonpregnant rats, and when partially delipidized, the lipoproteins from both groups showed similar immunoelectrophoretic characteristics and sedimentation rates. When lactation was interrupted during puerperium, serum lipoproteins returned to control levels; but in lactating rats, high levels of serum very low density lipoprotein persisted up to 3 wk post partum. PMID- 14564721 TI - Use of esters of N-hydroxysuccinimide in the synthesis of N-acylamino acids. AB - Several crystalline N-hydroxysuccinimide esters of short- and long-chain fatty acids have been synthesized. These compounds react with free amino acids to form preferentially N-acylamino acids. The reaction of the N-hydroxysuccinimide esters with hydroxylamine and the behavior of the N-acylamino acids on thin-layer chromatography are described. PMID- 14564722 TI - Crystallization of sodium taurocholate. AB - Success in crystallizing sodium taurocholate from ethanol by the addition of ether is critically dependent on the water content of the system. Two crystalline forms of sodium taurocholate were obtained with melting points of 180 degrees C and 225-235 degrees C respectively. PMID- 14564723 TI - Microreactor for methanolysis of triglycerides before gas-liquid chromatography. AB - A rapid, accurate microtechnique has been developed for gas chromatographic determination of the fatty acid composition of small (2-3 micro l) samples of vegetable oils. This microtechnique combines transesterification and sample injection into a single operation. The fatty acid compositions of soybean, linseed, and safflower oils thus determined are compared with those obtained by the usual two-step procedure. PMID- 14564724 TI - Separation of sterol acetates by column and thin-layer argentation chromatography. AB - Column and thin-layer chromatographic systems employing silver nitrate impregnated adsorbents are described for the separation of sterol acetates which differ in the number of double bonds in the steroid nucleus or side chain. PMID- 14564725 TI - Separation of 24- and 25-dehydrocholesterols; and the impure state of commercial desmosterol preparations. AB - Chromatographic methods for the separation of 24- and 25-dehydrocholesterols are described. The purities of three commercial samples of 24-dehydrocholesterol examined by thin-layer and gas-liquid chromatography were only 42, 79, and 80%, respectively; a commercial sample of radioactive 24-dehydrocholesterol was shown to contain 40% 25-dehydrocholesterol. PMID- 14564726 TI - Reversed-phase partition thin-layer chromatography of rat liver lecithins to yield eight simple phosphatidyl cholines. AB - The four fractions obtained by argentation thin-layer chromatography of intact rat liver lecithins can be further subdivided by reversed-phase partition thin layer chromatography on hydrophobic kieselguhr. The resultant eight fractions contain virtually only one saturated and one unsaturated acid each. PMID- 14564727 TI - Application of the Rosenmund reaction to the synthesis of saturated fatty aldehydes. AB - A method is described for the simple and rapid formation of saturated fatty aldehydes from the corresponding acid chlorides. It is not suitable for the preparation of unsaturated aldehydes because of the partial reduction and positional and geometrical isomerization of the double bond in the chain. PMID- 14564728 TI - Serum and lymph lipids in rabbits with carbon tetrachloride-induced cirrhosis of the liver. AB - Lymph flow and the composition of lymph lipids from the hepatic and thoracic ducts of rabbits with cirrhosis of the liver (induced by 46-51 intramuscular injections of a mixture of carbon tetrachloride and olive oil at 4-day intervals) have been compared with those of control animals injected with olive oil only. In cirrhotic animals, the concentration of lymph lipids was not greatly altered, but lymph flow, and consequently the hourly transport of lipids by lymph were greatly increased; the increase in transport of cholesteryl esters, free cholesterol, and phospholipids by way of the thoracic and hepatic duct lymph was particularly striking. The concentration of these lipid fractions in serum from the cirrhotic rabbits was also increased. The differences normally observed between lipid fatty acid compositions of serum and lymph disappeared in cirrhotic animals; this is interpreted as due to increased hepatic permeability to lipoproteins. PMID- 14564729 TI - Effect of light on extraction of lipid from retinal rods. AB - Chloroform-methanol 2:1 removes a significantly greater quantity of lipid from bleached bovine retinal rods than from a dark-adapted counterpart. The extracts contain phosphatidyl serine, phosphatidyl choline, phosphatidyl ethanolamine, sphingomyelin, and an unknown substance which, it is proposed, may be a combination of phospholipid and retinaldehyde. The difference between extracts of light-and dark-adapted rods is quantitative rather than qualitative. The data tend to confirm a model of rhodopsin suggested by Kropf and Hubbard in which isomerization of the retinaldehyde chromophore causes its displacement and opens a path to the interior of the molecule. PMID- 14564730 TI - Device for evaporation of solvent from small samples. AB - A device for the simultaneous evaporation of six samples under separately controlled streams of nitrogen is described. PMID- 14564732 TI - [Computer modeling the cytochrome P450 three-dimensional structure: problems and prospects]. AB - The review is devoted to the description of basic approaches, problems and prospects of computer modelling of 3D structures of cytochromes P450 (P450s). The following questions have been are considered in the review: (1) the role of computer molecular modelling in P450s research; (2) basic stages of homology modelling of protein 3D structures; (3) purposes and tasks of molecular modelling; (4) problems of P450s homology modelling; (5) criteria for successful P450s homology modelling; (6) methods of P450s models refinement, verification of correctness and reliability. PMID- 14564733 TI - [Analysis of the model binding site of anti-2,4-dichlophenoxyacetic acid antibodies]. AB - Models of three-dimensional structures of Fv domains of three antibodies specific to the pesticide 2,4-dichlorophenoxyacetic acid (2,4-D) have been derived from the sequence data by comparative modeling. The same binding site cavities were observed in all cases. The most important residue in antigen binding is tyrosine, which serves as a wall of cavity and putatively forms pi-stacking interaction with aromatic moiety of the ligand. Another cavity wall is formed by hydrophobic residues. At the entrance of cavity a glutamate residue is located in 2 of 3 structures. Docking of 2,4-D and its analogs on the models was performed. On the basis of docking results an experimental cross-reactivity data were qualitatively explained. Using results of the modeling, mutation of glutamate to serine or lysine was suggested to eliminate electrostatic repulsion between antibody and ligand and to improve 2,4-D binding efficiency. Target mutations in the antibody binding site were checked on the model. PMID- 14564734 TI - [Antioxidants in malignant neoplasms in humans and animals]. AB - This review is devoted to the role of low molecular weight antioxidants and antioxidant enzymes in cancerogenesis. The data on accumulation of lipid-soluble and water-soluble antioxidants in a number of malignant human and experimental tumors especially in the renal cell carcinoma tissues are analyzed. A possibility is discussed concerning the pro- and anticarcinogenic effects of the abundant tumor content of antioxidants. Implication of antioxidants in tumor resistance to chemotherapeutic drugs and ionizing radiation is discussed. PMID- 14564735 TI - [Lipid peroxidation in the spleen induced by chronic ionizing irradiation]. AB - Chronic gamma-radiation (1-16 sGr/day) of mice increased antioxidant activity and decreased lipid peroxidation. The was correlation between investigated parameters and the dose of irradiation. The results suggest, that chronic gamma-radiation causes damage of processes in spleen of CBA mice accompanied by subsequent augmentation of reparative processes. PMID- 14564736 TI - [Changes in lipid components of the rat brain gray matter caused Ca(2+) ions in postmortem autolysis in vitro]. AB - The influence of exogenous ions Ca2+ on the autholytic alterations of lipids in the brain grey matter of rats was investigated during incubation in a sterile physiological solution in vitro. The level of phospholipids decreased with parallel accumulation of diacylglycerols and free fatty acids. The contents of cholesterol and its esters varied in reciprocal oscillatory mode with prevalence of the disintegration of cholesterol esters started after 4 hrs of autholysis and further. The most probable degradation reactions of phospholipids and also cholesterol esters were the hydrolytic reactions. Transacylation reactions of between phospholipids, cholesterol and its esters were also possible. The presence of exogenous Ca2+ ions in the incubation medium promoted decrease of the total lipid grey matter. This effect was noted 10 min after onset of autolysis and continued up to 24 h. The reduction of lipid concentration occurred due to the decrease of phospholipid level and to a lesser extent--to the cholesterol level. PMID- 14564737 TI - [Glutathione-S-transferase activity in various brain regions of normal and fasting animals]. AB - Glutathione-S-transferase Y alpha Y alpha (GT-Y alpha Y alpha) and YbYb (GT-YbYb) activities were studied in cytosolic fraction of the cortical (limbic, orbital and sensorimotor cortex) and subcortical (myelencephalon, mesencephalon, hypothalamus) structures of rat brain under different terms of starvation (1, 2, 3, 5 and 7 days). In all brain structures GT-YbYb activities were approximately 2 times higher than those of the GT-Y alpha Y alpha activities. Basal activity of both forms of glutathione-S-transferase in cortical structures was also 2 times higher than that in the stem structures. Significant increase of activities of the GT-Y alpha Y alpha and GT-YbYb was selected both in cortical and subcortical structures on the third day and especially on the fifth day of starvation. It was concluded, different rat brain structures possess high glutathione-S-transferase activities, which are activated after long term starvation. PMID- 14564738 TI - [Effect of azole derivatives on lucigenin-dependent microsome chemiluminescence]. AB - Both metronidazole and aminotriazole increased while sanazole (drug AK-2123) decreased the NADPH/lucigenin-dependent chemiluminescence of liver microsomes of phenobarbital-treated rats. Sanazole strongly inhibited the lucigenin-dependent chemiluminescence in the enzyme system of xanthine-xanthine oxidase. Aminotriazole and metronidazole were less potent inhibitors of chemiluminescence less than sanazole. All these azole derivatives did not absorb light in the region of light emission of lucigenin. Both lucigenin and sanazole increased the rate of cytochrome c reduction by microsomes in case of using NADPH as a donor of electrons, whereas no effect of metronidazole and aminotriazole on this rate was found. The sanazole inhibition of lucigenin-dependent chemiluminescence could reflect competition between sanazole and lucigenin for electrons in the active centre of flavin reductases. Thus, microsomal NAD(P)H-reductases can be potentially involved in a bioactivation of sanazole. Lucigenin-dependent chemiluminescence cannot be used for measuring the modulating action of agents on reactive oxygen species production in the microsomes, but it may be used for luminometrical studies of enzyme complex NAD(P)H-reductases/cytochrome P450 in model systems. PMID- 14564739 TI - [Comparative effects of complement inhibitors in vitro and in vivo experiments: an immunoenzyme method in studying subcomponent C1q inhibition and complement inhibition in model animals]. AB - Methods of analysis of inhibition of complement system in vitro and in vivo have been developed for study of effects of medical drugs on the complement. The first one, ELISA method, for determination of inhibition of the first stage of complement activation includes binding of C1q subcomponent to immunoglobulin. The second method is based on capacity of mink serum to kill mice at the intravenous administration due to the action of mink complement. The effects of heparin, known anticoagulant, and suramin, used for treatment of trypanosomiasis, have been studied using these systems. The inhibition constants of binding suramin and heparin binding evaluated by the first method C1q were 411 +/- 29 micrograms/ml (or 0.287 +/- 0.020 mumole/l) and 36.4 +/- 1.7 micrograms/ml (or 2.28 +/- 0.10 mmole/l), respectively. This indicates that heparin binding with C1q in 10 times is higher, than that for suramin (as weight ratio) or 100 times higher in molar ratio. Administration of 3 mg of suramin or 0.3 mg of heparin to mice protected them against lethal action of intravenously injected 0.08 ml of mink serum. Blood concentrations of these compounds approximately correspond to inhibition constants for C1q binding, obtained using in vitro method. PMID- 14564740 TI - [Effect of a biologically active supplement (BAD) "SeleniumES" on the antioxidant system in patients with cardiovascular diseases]. AB - The use of the dietary supplement SeleniumEC resulted in the normalization of markers of the lipid peroxidation and the antioxidant system during complex diet therapy of patients with ischemic heart diseases. These markers were more stable in patients received SeleniumEC further one month after the period in the clinic. PMID- 14564742 TI - Professional sexual misconduct--an overview. PMID- 14564741 TI - [Distribution of chemical elements in whole blood and plasma]. AB - The distribution factor (Fd) of 35 elements of plasma and whole blood in 26 healthy men and women was detected by ICP-OES. Usilig this parameter the elements were subdivided in 3 pools. 9 of them have Fd higher than 1.5 ("elements of plasma"-Ag, Ca, Cu, In, Li, Na, Se, Si, Sr); 6 have lower than 0.5 ("elements of blood cells"-Fe, K, Mn, Ni, V, Zn), other 20-about 1 ("blood elements"). Fd of all elements depends on ionic radius. Elements of 2nd sub-groups of all groups of Mendeleev's periodic table ("heavy metals") depend on the similar law: "with growing of ionic radius the concentration of elements in plasma enhances". In alkaline metals Fd depends on the opposite law:" with growing of ionic radius of alkaline metal the quantity of elements in blood cells enhance". Dependence of Fd on the value of atomic mass in periods or in exterior electronic cloud (s-, p-, d , f-) was not established. The table of distribution of all detected elements in whole blood in relation to 8 macroelements (Ca, Mg, K, Na, S, P, Fe, Zn,) is presented, as a basic diagnostic criteria in metal-ligand homeostasis disturbance. PMID- 14564743 TI - "9-11" remembered. PMID- 14564744 TI - Looking back: Sluder method of tonsillectomy. PMID- 14564745 TI - Safety strategies to prevent suicide in multiple health care environments. AB - BACKGROUND: Patient suicide is one of the primary sentinel events reported throughout the United States. North Shore-Long Island Jewish Health System undertook a series of performance improvement efforts to identify suicide risk factors and develop a series of strategies and tools to maximize the safety of all vulnerable patients. METHODOLOGY: A multidisciplinary task force conducted root cause analyses of 17 attempted and completed suicides and targeted inadequate patient assessment, poor communication, and knowledge deficits. A protocol was designed to ensure appropriate assessment, monitoring, and treatment of patients at risk for alcohol withdrawal and suicide. Poor communication as patients moved throughout the continuum of care was addressed through targeted education, a centralized intake model, and an inter-institutional transfer summary form. A continuous suicide risk assessment tool was incorporated into the inpatient behavioral health rounds. SUMMARY AND CONCLUSIONS: The new tools have raised awareness, improved accountability, and encouraged best practices throughout the health system. PMID- 14564746 TI - One hospital's journey toward reducing medication errors. AB - BACKGROUND: The Valley Hospital, a 451-bed acute care facility in Ridgewood, New Jersey, has made substantial progress in the reduction of medication administration errors. METHODS: Reduction in medication administration errors were accomplished through (1) becoming intimately familiar with the errors, including where, when, why, and how they were occurring; (2) establishing a nonpunitive environment and encouraging reporting of errors, including near-miss errors; (3) trending error report data to identify areas of concentrated errors in the medication use process; (4) simplifying and standardizing process steps; and (5) selecting the right technology to address error-prone steps in the hospital's systems. RESULTS: The establishment of a nonpunitive environment led to a dramatic increase in the number of nearmiss errors reported, and the information gained proved to be valuable and diagnostic. Establishing an interview process with the caregivers directly involved in occurrences enabled us to gather detailed information about errors. This forum led the way to an early understanding of human factors, system failures, and root cause analysis. Those errors were trended, addressed, and reduced through manual system changes and technological system developments designed to ensure the "five rights" of safe medication administration. CONCLUSIONS: Keeping on course requires constant and continuous review of medication use processes to ensure that they support instead of unnecessarily limit actual practices. PMID- 14564747 TI - Characteristics of physician leaders working to improve the quality of care in acute myocardial infarction. AB - BACKGROUND: The influence of physician leaders on their colleagues in local medical communities has been recognized for several decades. However, the literature indicates that little is known about the specific characteristics of physician leaders involved in improving quality in today's hospital environment. A taxonomy of the characteristics of the physician quality leader from the perspective of physicians and nonphysicians was developed. SUBJECTS AND METHODS: Information about physician leaders working to improve acute myocardial infarction (AMI) was gathered from in-depth interviews with 45 key physicians and nursing, quality management, and administrative staff at eight hospitals. Data were analyzed using the constant comparative method of qualitative data analysis. RESULTS: The physician leader characteristics were described in four main categories: personal commitment, professional credibility, quality improvement behaviors and skills, and institutional linkages. Each physician leader possessed different combinations of the characteristics from the four categories, revealing the complexity of the physician leader role. CONCLUSION: Understanding the key characteristics of physician leaders is a critical step in helping hospitals choose and develop physician leaders who can effectively bring about meaningful quality improvement. PMID- 14564748 TI - Microsystems in health care: Part 5. How leaders are leading. AB - BACKGROUND: Leading and leadership by formal and informal leaders goes on at all levels of microsystems--the essential building blocks of all health systems--and between them. It goes on between microsystems and other levels of the systems in health care. This series on high-performing clinical microsystems is based on interviews and site visits to 20 clinical microsystems in the United States. This fifth article in the series describes how leaders contribute to the performance of those microsystems. ANALYSIS OF INTERVIEWS: Interviews of leaders and staff members offer a rich understanding of the three core processes of leading. Building knowledge requires many behaviors of leaders and has many manifestations as leaders seek to build knowledge about the structure, processes, and patterns of work in their clinical microsystems. Taking action covers many different behaviors--making things happen, executing plans, making good on intentions. It focuses action on the way people are hired and developed and involves the way the work gets done. Reviewing and reflecting provides insight as to how the microsystem's patterns, processes, and structure enable the desired work to get done; what success looks like; and what will be next after that "success" is created. CONCLUSION: The focus on the processes of leading is intended to enable more people to develop into leaders and more people to share the roles of leading. PMID- 14564749 TI - What is the best way to schedule patient follow-up appointments? AB - BACKGROUND: What is the best way to schedule follow-up appointments? The most popular model requires the patient to negotiate a follow-up appointment time on leaving the office. This process accounts for the majority of follow-up patient scheduling. There are circumstances when this immediate appointment arrangement is not possible, however. The two common processes used to contact patients for follow-up appointments after they have left the office are the postcard reminder method and the prescheduled appointment method. METHODS: In 2001 the two methods used to contact patients for follow-up appointments after they had left the clinic were used for all 2,116 reappointment patients at an ophthalmology practice at Dartmouth-Hitchcock Medical Center. The number of completed successful appointments, the no-show rate, and patient satisfaction for each method were calculated. RESULTS: A larger number of patient reappointments were completed using the prescheduled appointment procedure than the postcard reminder system (74% vs 54%). The difference between completed and pending appointments (minus no-shows) of the two methods equaled 163 patients per quarter, or 652 patients per year. Additional revenues associated with use of the prescheduled appointment letter method were estimated at $594,600 for 3 years. SUMMARY: Using the prescheduled appointment method with a patient notification letter is advised when patients do not schedule their appointments on the way out of the office. PMID- 14564750 TI - The evacuation of lower Manhattan by water transport on September 11: an unplanned "success". PMID- 14564751 TI - Making room for "outside the box" thinking in emergency management and preparedness. PMID- 14564752 TI - Using a stage model of behavior change to prompt action in an immunization project. PMID- 14564753 TI - Contact lenses. Preface. PMID- 14564754 TI - Extended wear contact lenses. AB - The experience with the complications associated with extended wear contact lenses has made practitioners and patients fearful of this modality. Today, strong scientific evidence suggest that the hypoxia problems associated with extended wear lenses have been solved. Good clinical evidence suggests that the new high-Dk silicone lenses are safe and can be worn successfully for up to 30 days. Patients can now be offered contact lenses that meet their desire of continuous vision; however, practitioners and patients must be instructed regarding the signs and symptoms of contact lens complications. Practitioners need to be educated in the diagnosis and management of the complications. Patients need to be educated about the need for proper eye care, contact lens care regimens, and appropriate follow-up visits. PMID- 14564755 TI - The effects of low- and hyper-Dk contact lenses on corneal epithelial homeostasis. AB - Contact lens were and eyelid suturing diminish overall Bcl-2 expression in the corneal epithelium, yet nuclear Bcl-2 in the exposed surface epithelial cells seems to be retained for longer periods when compared with the retention in control eyes. This observation may explain why there is a decrease in surface cell exfoliation during CTL wear, because nuclear Bcl-2 loss needs to precede apoptotic surface cell exfoliation. In addition, if the hypothesis is true that Bcl-2 also has a regulatory function in the proliferation and differentiation of tissues, the overall reduced expression of Bcl-2 (nuclear plus cytoplasmic) may clarify the decrease in proliferation of the corneal epithelium seen during CTL wear. Future studies should clarify the role of Bcl-2 in the homeostatic dynamics of the corneal epithelium. CTL wear and eyelid suturing provide excellent noninflammatory models to advance knowledge of Bcl-2 and its regulatory roles. Additionally, these models should help researchers gain a better understanding of the CTL-associated effects on corneal epithelium. PMID- 14564757 TI - Disposable and frequent replacement contact lenses. AB - Disposable and frequent replacement contact lenses dominate the marketplace. They are available in a wide variety of parameters for use in refractive errors, including myopia, hyperopia, astigmatism, and presbyopia. They are convenient and affordable and lend themselves to most wearing modalities, including daily wear, flexible wear, and extended wear for up to 30 days. In addition, they have been shown to be the most trouble-free contact lens wear modality for daily wear or extended wear. They are an appropriate choice for patients who desire occasional contact lens wear and have gained wide acceptance as therapeutic bandage contact lenses. Disposable and frequent replacement lenses will remain important modalities for some time to come as the variety of contact lenses and contact lens parameters that are offered continue to expand. New contact lens varieties, such as the high-Dk silicone hydrogel lenses, will further expand the role of these contact lenses in vision correction. PMID- 14564756 TI - Corneal hypoxia secondary to contact lenses: the effect of high-Dk lenses. AB - Contact lenses made from materials of low-oxygen permeability (Dk) do not meet the oxygen requirements of the cornea for overnight wear. Long-term extended wear of these lenses results in chronic changes to all layers of the cornea, many of which are associated with hypoxia. High-Dk silicone hydrogel and gas permeable lenses are now available for 30-night continuous wear. The high-oxygen transmissibilities of these lenses have enabled the development of a successful continuous wear modality by eliminating the hypoxic effects of long-term wear. Presently, the focus is on improving lens performance by developing lenses that are more biocompatible, provide greater comfort, and maintain a stable tear film without inflammatory or mechanically induced adverse events. PMID- 14564758 TI - The correction of astigmatism with soft contact lenses. AB - Soft contact lens correction for astigmatism has made significant advances over the last 20 years. Soft toric tinted lenses, disposables lenses, and bifocal lenses are now available at lower costs with greater reproducibility, enhanced parameters, and better comfort. Because of these innovations and significant design changes, a greater percentage of astigmatic patients are being treated with soft toric lenses than before. PMID- 14564759 TI - Correction of astigmatism with rigid gas permeable lenses. AB - Comparison of the results of keratometry and refraction usually suggests the appropriate contact lens for a patient's eye. Specialized rigid toric contact lenses include bitoric and back toric RGP lenses, front toric RGP lenses, and bitoric RGP lenses with prism ballast. Each lens has specific indications. PMID- 14564760 TI - Presbyopic correction with contact lenses. AB - According to Paragon Vision Sciences (Mesa, AZ), an additional 50,000 Americans become presbyopic each day. There are many contact lens options available to help these patients meet their visual needs in the workplace and for recreational and social activities. Although technology still requires some compromise on the part of many patients, constant advances in materials and designs should enable even greater numbers of presbyopes to enjoy the freedom of functioning without glasses in the years to come. PMID- 14564761 TI - Use of contact lenses for management of keratoconus. AB - This article describes use of contact lenses to manage patients with keratoconus. Fitting concepts, prefit evaluations, types of lenses, initial lens selection, and potential problems and solutions are discussed. PMID- 14564762 TI - Contact lens management following corneal refractive surgery. AB - Corneal refractive surgery has enjoyed considerable success and popularity in recent years; LASIK has become the surgical procedure of choice. The incidence of visually significant complications varies widely, but is inversely related to surgeon experience. Visual complaints include multifocal images, shadows, ametropias, anisometropia, night vision difficulties, decreased contrast sensitivity and loss of BCVA. Contact lenses offer an option to rehabilitate patients when visually symptomatic following refractive surgery. RGP lenses provide the best postoperative visual rehabilitation following refractive surgery. PMID- 14564763 TI - Modern corneal reshaping with contact lenses. AB - The renaissance of corneal reshaping has been influenced dramatically by new lens designs, high-oxygen permeable materials, and corneal mapping techniques. These evolutionary advances have dramatically changed the practitioner's view of the modality. The procedure has evolved from a specialty practiced by only a handful of practitioners to a technique that can be incorporated successfully into almost any eye care practice. PMID- 14564764 TI - Current contact lens care systems. AB - Contact lens care has been revolutionized and simplified with the development of multipurpose solutions, less sensitizing preservatives and disinfectants, better protein removers, and reaction-free, one-bottle care systems for patients who have had adverse responses to existing care products. As the complexity of lens care has decreased, compliance has increased. Nevertheless, good compliance is dependent on technician training and a thorough understanding of the chemistry and mechanism of action of each care system. Care products are constantly evolving, and the practitioner should become familiar with each new solution as it is introduced to ensure that patients are also kept up-to-date. PMID- 14564765 TI - Corneal topography and contact lenses. AB - Regardless of whether CVK is used qualitatively for RGP lens design selection or quantitatively in RGP parameter selection, it has a significant role in contact lens practice. Further advancements and testing on CVK-based RGP fitting modules and fluorescein simulations will make CVK an invaluable tool for the RGP lens fitter. Soft lens applications of CVK data are already being tested [42,43]. This technology will become the standard of care for all contact lens patients and will most likely replace the keratometer. PMID- 14564766 TI - Therapeutic contact lenses: the role of high-Dk lenses. AB - Currently, the armamentarium of contact lenses that can be used for therapeutic effect provides a wider selection of lenses than ever before. If the therapeutic goal is protection and healing of the corneal epithelium, epithelial or stromal edema is best avoided, and the selection of a high-Dk silicone hydrogel (balafilcon A, lotrafilcon A) lens or a very thin membrane-type lens (crofilcon) is the best choice. If the goal is surface protection as well as stimulation of stromal wound vascularization, selection of a low-water content, thick, hydrophilic lens is the better option. If the patient is prone to lens loss or requires frequent replacement of the therapeutic lens, a prudent economic decision is to select a daily disposable moderate-water content lens. Specific circumstances may mandate the selection of a specific therapeutic lens. Patients with a prior history of active giant papillary conjunctivitis may be better served by the use of a crofilcon glyceryl methacrylate lens, which has a lower incidence of this complication. Patients who have dry eye may benefit from a higher-water content lens if adequate unpreserved tear supplementation is provided with or without punctal occlusion. The options when selecting a therapeutic contact lens are wider than ever before. Although the new generation of high-Dk lenses promises fewer limiting problems of vascularization and infection, one can use the older traditional therapeutic lenses when induced vascularization of the cornea is needed or when an economic necessity exists. Not all of the available lenses are FDA approved for therapeutic use, and such wear is an off-label use. The patient should be informed of the goal of therapy as well as the benefits and risks of therapeutic contact lenses. PMID- 14564767 TI - Contact lenses and associated anterior segment disorders: dry eye, blepharitis, and allergy. AB - Contact lenses should be used with caution in patients who have an ocular allergy. Patients who have seasonal allergy should avoid contact lens use during seasonal flare-ups. The need for clean lenses with minimal deposit buildup must be stressed, and the use of daily wear lenses with rigid disinfecting and cleaning techniques is recommended. Alternatively, daily disposable lenses should be used. The patient should avoid the use of topical anti-allergy agents while the lenses are in place, particularly vasoconstrictor agents. Extended wear contact lenses are contraindicated in patients who have ocular allergy. In general, the use of contact lenses is contraindicated in patients with vernal conjunctivitis. With careful attention to recognizing the patient with ocular allergy, regular monitoring, and patient compliance to lens care, successful contact lens wear can be achieved in most patients with ocular allergy. PMID- 14564768 TI - Contact lens complications. AB - Complications associated with contact lenses range from mild to severe and occur with all lens modalities. Contact lens wear can cause a change in corneal physiology, which can lead to epithelial, stromal, and endothelial compromise. Other complications include lens deposition, allergic conjunctivitis, giant papillary conjunctivitis, peripheral infiltrates, microbial keratitis, and neovascularization. Pre-existing conditions can contribute to these complications, or they can occur in association with contact lens wear and care regimens. Patient-related factors, such as alteration of the recommended wearing or replacement schedules and noncompliance with recommended contact lens care regimens for economic reasons, convenience, or in error, contribute to contact lens-related complications and have led to difficulty in accurate determination of complication rates among the various lens wear modalities. Complications may require discontinuation of contact lenses, topical therapy, and changes in contact lens wearing schedules, materials, and care solutions. On initial lens fitting and follow-up evaluations, practitioners should review contact lens replacement and cleaning regimens with patients and discuss complications. To avoid serious complications, patients should be reminded to remove their contact lenses as soon as ocular irritation occurs, and to call their eye care practitioner immediately if symptoms persist. PMID- 14564769 TI - Present status of contact lens-induced corneal infections. AB - This article addresses the spectrum of corneal infections associated with contact lens wear. The status of bacterial, fungal, and Acanthamoeba keratitis is discussed as well as diagnosis and treatment for these infections. PMID- 14564770 TI - [Structural-dynamic analysis of the mechanism of clinical response and treatment prognosis]. AB - The paper presents the results of testing of a new approach to study the inter syndrome links in initial and follow up status of patients treated with atypical antipsychotics and to determine a system of prognostic clinical factors. The method, named structural-dynamic analysis, is based on comparison of initial and final structure of patient's condition grouped by the features of dynamics of its separate elements. These elements are chosen in accordance to PANSS and the grouping, therefore, is performed 30 times. The subject of the study was a sample of 89 patients with acute exacerbation of schizophrenia with intermittent course treated with atypical antipsychotic as monotherapy. The results of the study demonstrated that the new approach was informative and allowed to elucidate the most important parts of patient's condition, changes of which served as a basis for alterations in many other aspects of clinical status and consequently could be determined as the main characteristics of treatment efficacy. At the same time, these parts not in all the cases define the features of the clinical picture, being overlapped by the more striking symptoms. However, changes of the latter much less reflect the global changes in biological processes influenced by antipsychotics. PMID- 14564771 TI - [Blood flow insufficiency in vertebrobasilar system]. AB - Ethiopathological and clinical factors of vertebral blood flow insufficiency development are discussed in the article. Distinct connection between pathogenesis and stages of clinical manifestation is proved and nosological independence of vertebrobasilar disease is validated. Used in combined therapy of patients with the disease, cavinton is shown to exert a positive effect on the main clinical symptoms. PMID- 14564772 TI - [Subjective body experience in hypochondria and somatoform disorders]. AB - Limitations and difficulties of clinical characteristics of mental disorders, manifesting in symptoms of body sphere, are described. The conceptions of body and subjective body experience, assuming that body sensations are involved in self-identification and self-ego formation and connected with cognitive, social and cultural aspects of subjective experience, are discussed. To evaluate phenomenological, biographical, psychodynamic, cognitive and socio-cultural context of body sensations appearance in somatoform mental disorders, the multi axial model of subjective body experience description is suggested. PMID- 14564773 TI - [Efficacy of acupressure therapy in combined treatment of psycho-autonomic neurotic disorders in children]. AB - The aim of the study was to evaluate the efficacy of acupuncture in combined treatment of psychoautonomic neurotic disorders in children. 163 children, mean age 12.5 years, were included in the study. Autonomic dysfunction score, clinoorthostatic test, sinocarotid test and heart rate variability analysis were used to assess the efficacy of different therapeutic strategies applied: complex (acupuncture + conventional drug treatment)--group 1; acupuncture treatment- group 2, and conventional drug treatment--group 3. After 2 months of treatment, a reduction of autonomic dysfunction score and an increase of percentage of patients with normal clinoorthostatic and sinocarotid tests were revealed in all the groups, being most evident in group 1. Heart rate spectral variability changed in patients given acupuncture treatment (groups 1 and 2). Relative augmentation of sympathetic activity was observed in patients with initial vagotonia, while those with initial symphaticotonia exhibited a relative parasympathetic activity increase. Thus, acupuncture exerts beneficial effects on functioning of autonomic nervous system. The method may be used either independently, or in addition to conventional drug treatment of psychoautonomic neurotic disorders in children. PMID- 14564774 TI - [Clinical efficacy and tolerability of tianeptine in treatment of depression (Russian open multicenter trial)]. AB - One hundred and forty two patients with ICD-10 diagnosis of depression from 7 research scientific were treated with tianeptine (coaxil); 124 have completed the treatment course. Moderate depression was diagnosed in 111 patients (89.5%) and severe depression--in 13 (10.5%). A duration of the last episode before treatment was 7.1 +/- 1.7 months. The mean score by Hamilton depression scale was 24.4 +/- 4.0. The patients received tianeptine as a monotherapy in dosage 12.5 mg, 3 times daily during 6 weeks (65-year olds and older were given 25 mg daily). Tianeptine proved to be an antidepressant with balanced action, i.e., exerted thymoanaleptic, anxiolytic and activating effects. The treatment was beneficial for 70.4% of the patients; remission was revealed in 58%. The medication was well tolerated with rare and weakly pronounced side effects. PMID- 14564775 TI - [The experience of pyrazidol use in the treatment of non-psychotic depression]. AB - Pirazidolum ("Masterlek", Russia) was used in the treatment of 30 patients with recurrent depression (ICD-10 item F33) and depression episode (F32), mild and moderate types. The initial dose was 50-75 mg daily, mean dose--200-350 mg daily, treatment course--28 days. The efficacy of the treatment was evaluated by Clinical Global Impression (CGI) scale, Hamilton depression (HAM-D) scale and side effects scale (UKU). Significant improvement was revealed in 73% of the patients. A trend to improvement scored on HAM-D was detected by 2 treatment week 2, and pronounced improvement (more than a half-decreased total score)--by week 4. Only 27.6% of the patients had weak side effects (dry mouth, sweatiness, tachycardia etc) that did not demand any changes in the treatment course. The authors believe that pirazidolum may be referred to antidepressants which are effective for depression with a prevalent positive affect. PMID- 14564776 TI - [Spectrum of therapeutic efficacy and safety of Lorafen (lorazepam) use for anxiety disorders]. AB - Presented here, are the results of 4-weeks lorafen monotherapy in 31 in-patients of psychiatric and cardio-surgical clinics divided, respectively, into 2 groups: 16 patients with chronic anxiety disorders and 15--with acute developed before an coronary artery bypass surgery in ischemic heart disease. A dynamics of the patients' state was evaluated with Hamilton Rating Scale for Anxiety and the Hospital Anxiety and Depression Scale. Lorafen proved to be highly effective, with percentage of responders being about 77% (24 patients out of 31). Therapeutically effective dosage was 3.7 +/- 1.1 mg daily for chronic anxiety disorders and 2.1 +/- 0.6 mg--for situation-induced anxiety. Lorafen was shown to be tolerable and safety. PMID- 14564777 TI - [Cognitive potential p300 in Parkinson disease]. AB - Cognitive evoked potential P300 in response to visual verbal and non-verbal stimuli was examined in 40 patients with idiopathic Parkinson's disease (PD) without cognitive impairment and 8 age- and sex matched normal controls. All the patients were subjected to clinical and neuropsychological evaluation. The influence of patient's age, duration, type and severity of the disease, debut's lateralization and anti-parkinsonic therapy efficacy on P300 was evaluated. Compared to controls, PD patients exhibited a significantly prolonged P300 latency and decreased P300 peak amplitude. The topography of PD maximum P300 amplitude indicated a frontal distribution with no between-hemispheres differences. Overall, the P300 changes in PD patients without dementia can be interpreted as a decrease of directed activity level and the partial defect of mental recognition processes and stimuli differentiation related mostly to the nonverbal image-spatial cognitive processes. The results show that P300 recording may be useful for diagnosis of subclinical cognitive disturbances in PD without dementia. PMID- 14564778 TI - [Diagnostic algorithm for upper extremity peripheral paresis]. AB - The authors have rejected applying of image identification algorithms and searching for given sequence in favor of algorithms sorting, eliminating thereby an influence of subjective factors on diagnostic process. To create the program, the diagnostic signs with probability coefficient 1 (positive indicator) or 0 (negative indicator) were used. To positive signs were attributed indicator muscles and indicator sensitive zones--the cutaneous areas where sensitive disorders appeared in the first turn, in response to a lesion in corresponding segment of peripheral nervous system, and disappeared in the last turn; to negative ones--muscles or cutaneous areas, being never affected by paresis or hyperesthesia in response to the given lesion type. The first part of the algorithm concerns a definition of defect level and the second one--an establishment of final diagnosis. The use of the algorithm developed simplifies diagnostics and raises its efficacy. PMID- 14564779 TI - [Application of a mathematical algorithm for the detection of electroneuromyographic results in the pathogenesis study of facial dyskinesia]. AB - To carry out a differential diagnosis of two facial dyskinesia (FD) models- facial hemispasm (FH) and facial paraspasm (FP), a combined program of electroneuromyographic (ENMG) examination has been created, using statistical analyses, including that for objects identification based on hybrid neural network with the application of adaptive fuzzy logic method and standard statistics programs (Wilcoxon, Student statistics). In FH, a lesion of peripheral facial neuromotor apparatus with augmentation of functions of inter-neurons in segmental and upper segmental stem levels predominated. In FP, primary afferent strengthening in mimic muscles was accompanied by increased motor neurons activity and reciprocal augmentation of inter-neurons, inhibiting motor portion of V pair. Mathematical algorithm for ENMG results recognition worked out in the study provides a precise differentiation of two FD models and opens possibilities for differential diagnosis of other facial motor disorders. PMID- 14564780 TI - [Particular aspects of bioelectric brain activity in post-traumatic Korsakoff syndrome]. AB - Thirty two patients, 21 with recurrent and 11 with chronic Korsakoff's syndrome of traumatic genesis, and 20 healthy subjects were studied, using visual and spectral coherent EEG analysis and investigation of three-dimensional localization of dipolar beta 1-activity sources (13-20 Hz EEG bands). In Korsakoff's syndrome, negative prognostic significance of a stable increase of EEG beta 1-activity coherence generated by a limited number of sources, preferably of frontal basal and stem localization, was found. In the context of clinical data, this EEG phenomenon may be related to inhibited state of corresponding cortical areas. PMID- 14564781 TI - [Proximal autosomal recessive types of spinal muscular atrophy]. AB - One hundred and three patients with spinal muscular atrophy (SMA) were registered in a population medical genetic study of autosomal recessive childhood proximal SMA in Saratov region. Twenty-five patients were investigated complexly, using biochemical analysis of some enzymes, electroneuromyography, magnetic resonance imaging of spinal cord, muscle biopsy and molecular genetic testing. Pronounced clinical polymorphism and genetic heterogeneity of the disease were revealed. PMID- 14564782 TI - [A comparison of some indices of innate and adaptive immunity in different types of schizophrenia]. AB - Leukocyte elastase (LE) blood serum activity as an index of innate immunity and autoimmune reactions to brain antigens i.e. a level of autoantiboies to nerve growth factor (NGF) as an index of adaptive immunity were studied in patients with attack-like and slow progressive schizophrenia. Compared to controls, higher LE activity accompanied by a significant increase of autoantibodies to NGF titers was found in patients with attack-like schizophrenia. In slow progressive schizophrenia, only higher LE activity was detected. Correlations between immunological parameters and some clinical appearances (positive and negative disorders) and disease course peculiarities were revealed. The results suggest an involvement of different parts of immune system in pathophysiology of attack-like and slow progressive schizophrenia. PMID- 14564783 TI - [Positron emission tomography in patients with obsessive-compulsive disorders of a hereditary type]. PMID- 14564784 TI - [History of neurology in the Dobass region]. PMID- 14564785 TI - [The experience of dopaminergic agent decreased dosage with rhythmic pace gait correction in Parkinson disease]. PMID- 14564786 TI - [Status of free radical lipid peroxidation in spinal fluid in chronic inorganic phosphorus compounds neuro-intoxication]. PMID- 14564787 TI - [Ontogenic aspects of depressive disorders]. PMID- 14564788 TI - [Opiate and opioid tolerance: toxico-kinetic and toxico-dynamic aspects]. PMID- 14564789 TI - Ninth circuit upholds award on resident's retaliatory discharge claim. Ostad v. Oregon Health Sciences University. PMID- 14564790 TI - Hospital secures injunction to discharge patient. Wyckoff Heights Medical Center v. Rodriguez. PMID- 14564791 TI - [Cervical fractures in autopsy records]. AB - We reviewed the autopsy records of 1872 cases of death because of politrauma, gunshot wounds and suicidal hanging. The analysis included causes and frequency of cervical spine fractures, their most common localisation, architecture of bone destruction and their influence on cervical cord. The most common cause of cervical spine injury was motor vehicle accidents. We examined 82 specimens with traumatic fractures of cervical spine obtained from accident victims. About half of the injuries occurred in upper cervical spine. The most common fracture localisation was C2 with dens fracture as the most frequent injury. The most common spinal cord lesion was complete rupture mainly at the upper cervical spine level. PMID- 14564792 TI - [Long-term results of revisional hip alloplasty using the X-change technique]. AB - In this paper the authors evaluate the long-term results of revisional hip alloplasty using the X-change technique with an Exeter endoprosthesis. This evaluation is performed using clinical material consisting of 52 operated patients, namely 41 female patients (78.8%) and 11 male patients (21.2%). In order to evaluate achieved results the authors propose such time criteria that performed remodelling of implanted bone grafts allows for full weight-bearing on the operated limb. They emphasize the importance of well-performed operation according to the rules of X-change technique, the way of implanting homogenous frozen bone and the way of cementing the implant, which all are indispensable to the success of the operation. Following those steps, the authors have achieved full remodelling of bone grafts between 12 and 16 month since the day of the operation. PMID- 14564793 TI - [Damage of hip prosthesis causing failure in arthroplasty]. AB - Analysis of retrieved hip joint was presented. Bone remodeling considering the distribution of friction products was discussed. Scanning electron microscopy methods were used to perform on structural analysis. We expressed a surface changing prosthetic head as Ra (roughness average). X-Ray Microanalysis was applied to study products of degradation. PMID- 14564794 TI - [Treatment of degenerative disease of knee joint with the Oxford prosthesis]. AB - Eleven patients with single compartment arthrosis of the knee were treated surgically, using the Oxford prosthesis. The age of the patients at the time of surgery ranged from 48 to 73 years. Varus deformity of the knee before surgery ranged from 0-23 degrees (average: 11 degrees). Patients with arthrosis of the medial compartment were elected for surgery. Surgery was performed only when passive correction of the varus deformity was possible and when the cruciate ligaments were intact. In all cases surgery yielded a stable knee joint. PMID- 14564795 TI - [Closed reduction and pinning for acute slipped capital femoral epiphysis]. AB - Acute slipped capital femoral epiphysis is a rare adolescent hip disorder, which may be a problem for orthopedic surgeon. No series to date has demonstrated the superiority of any treatment method. The purpose of the study was to evaluate the outcome of treatment and to assess the risk factors of avascular necrosis associated with this condition. Fourteen cases of acute slipped capital femoral epiphysis were treated with closed reduction and pinning. There were 9 boys and 5 girls. The average age at presentation was 11.8 years (range 9.1-15.3). Eight children were treated 4-10 days and six within 48 hours of the onset of their acute symptoms. The severity of the slip was classified according to the system of Southwick. Presence of chondrolysis and avascular necrosis was estimated. The clinical results were graded with the use of the criteria of Aadelen et al. Four slips were classified as mild, seven as moderate and three as severe. Average follow up was 5.2 years (range from 2.4 to 14.3). In two hips with severe slip AVN developed. One of these cases was treated within 48 hours and one after 7 days. Chondrolysis developed in one hip and was associated with presence of intra articular fixation. Twelve of the fourteen hips had good and vary good results and remaining 2 with AVN had poor. Closed reduction and pinning in acute slipped capital femoral epiphysis is an effective and giving satisfactory outcome method of treatment. Early reduction is not associated with a high rate of AVN and should be performed as soon as possible. More severe slips were noted to have an increased risk of AVN. PMID- 14564796 TI - [Fractures of the tibia's intercondylar eminentia in children]. AB - We have described management in tibia's intercondylar eminentia fractures in children, based on own data and review of the contemporary bibliography. The data contain 16 children from 6 to 14 years of age with all Meyers' and McKeever' fractures types. Type I and II fractures can be treated conservatively. Such factors like fracture's type and child's age must be taken under consideration for an operative treatment most often, but in children below 10 years of age, the conservative one should be considered as well. PMID- 14564797 TI - [Long-term results of treatment for articular calcaneus fractures with Westhues method]. AB - In this paper the authors evaluate the long-term results of treatment for articular calcaneus fractures with Westhues method. This evaluation is performed with regard to triple measurement of Bohler's angle value and to subjective criteria. The clinical material from years 1985-2002 consists of 47 patients aged between 24 and 56 years (mean age 32 years) with articular calcaneus fractures- namely, 41 male patients (87.2%) and 6 female patients (12.8%). The follow-up examination was performed in 24 patients (51% of all patients), namely, in 23 male patients (48.8%) and in one female patient (2.2%). Minimum follow-up time was 12 months and maximum was 9 years. Acquired results suggest that Westhues method restores normal Bohler's angle value only in tongue-type calcaneous fractures. PMID- 14564798 TI - [Evaluation of the effectiveness of the calcaneal spurs radiotherapy]. AB - The radiotherapy results of 46 calcaneal spurs were analysed. Patients were irradiated using fraction dose of 1.0 Gy, 1.5 Gy or 2.0 Gy up to total dose of 2.0-22.5 Gy. All patients previously had chronic pain in the plantar region influencing possibility of walking and not reacting for conventional treatment. Whole group was periodically controlled after the treatment completion. The mean follow-up period was 12.5 months. The complete pain relief was observed in 9% cases one month after treatment and in 52% during the last control. The lack of analgetic effect was noted only in 3% cases as well one month after the treatment as during the last examination. Performed statistical analysis showed only inverse significant correlation between degree of pain relief six months after the treatment and duration of symptoms. Obtained results allow to form conclusion that radiotherapy of calcaneal spurs is easy, effective and safe way of analgetic treatment giving good results after low fraction and total delivered doses and that its efficacy depends on symptoms duration what is correlated to disease stage. PMID- 14564799 TI - [Changes in bone strength during convalescence after immobilization induces bone loss--experiment with adult rats]. AB - The aim of the present study was to evaluate the impact of a hindlimb immobilization on mechanical properties of mature bone, both cancellous and cortical tissue. The effectiveness of bone recovery during free activity after remobilization was also studied. The right hindlimb of mature 24 weeks old male rats was immobilized for three weeks. Femora and proximal tibiae methaphyses as well as the straight muscle of thigh were investigated post mortem just after immobilization period or four weeks after remobilization. Mass of the muscle, mass, density and mineralization of bones as well as mechanical properties in axial compression (tibiae metaphyses) or in three-point bending (femora) were measured. After immobilization all parameters were significantly lower in immobilized tibia than in the left, loaded and in respect to the right tibia in the control group. In femur a decrease of dry mass was observed, without any change of breaking force and work to fracture. After four weeks of convalescence, a significant worsening of mechanical competence of femur, and no recovery in tibia was stated. The results obtained indicate that a restoration of mobility without any additional rehabilitation is not enough to stop the processes of bone deterioration induced by unloading in mature bone. PMID- 14564800 TI - [The head-body index used to access femoral head size]. AB - Basing on an analysis of 260 X-rays of healthy hip joints in children and adults the authors present a head-body index (according to Kruczynski), used to assess femoral head size. The index is defined as the ratio of the circumference of circle drawn around the femoral head to the body width of the femur (measured below the minor trochanter). In patients less than 17 years old the index was 170 (SD-14.6). PMID- 14564801 TI - [Results of treatment of displaced long bones fractures in children--comparison of different methods]. AB - Authors compared functional results of treatment of 48 patients; 25 children with displaced supracondylar humeral fractures, 16 treated with skeletal traction (group I, mean age 7.9 year), 9 treated with closed reduction and percutaneous K wire fixation (group II mean age 7.7 year), and 23 patients with displaced femoral shaft fractures, 10 treated with traction (group III, mean age 7.5 year), 13 treated with closed reduction and elastic intramedullary nailing. Good results after supracondylar humeral fractures were obtained in 37.5% of patients treated with traction and in 66.7% treated with percutaneous K-wire fixation, and after femoral fractures in 70% treated with traction and 84.6% treated with intramedullary nailing. The use of instrumental methods of treatment allowed to reduce significantly the duration of hospitalization (respectively: 19.8 vs. 6.4, and 28.5 vs. 9.5 days). In authors' opinion, a closed reduction secured by percutaneous K-wire fixation as well as a closed reduction with elastic intramedullary nailing are the methods of choice in treatment of discussed fractures in children. PMID- 14564802 TI - Clinical alarms goal remains intact.... Joint Commission expands 2004 goals. PMID- 14564803 TI - Meeting the challenge: how hospitals complied with the new clinical alarms requirement. PMID- 14564804 TI - Breaking out of the biomed box: an audit assessment and recommendations for an in house biomedical engineering program. AB - In order to assess the current performance and to identify future growth opportunities of an in-house biomedical engineering (BME) program, senior management of Lehigh Valley Hospital (Allentown, Penn) engaged (in July 2001) the services of a clinical engineering consultant. Although the current in-house program was both functionally and financially sound, an independent audit had not been performed in over 4 years, and there were growing concerns by the BME staff related to the department's future leadership and long-term support from senior management. After an initial 2-month audit of the existing program, the consultant presented 41 separate recommendations for management's consideration. In order to refine and implement these recommendations, 5 separate committees were established to further evaluate a consolidated version of them, with the consultant acting as the facilitator for each group. Outcomes from each of the committees were used in the development of a formal business plan, which, upon full implementation, would not only strengthen and refine the current in-house service model but could also result in a substantial 3-year cost savings for the organization ($1,100,000 from existing operations, $500,000 in cost avoidance by in-sourcing postwarranty support of future capital equipment acquisitions). Another key outcome of the project was related to the development of a new master policy, titled the "Medical Equipment Management Program," complete with a newly defined state-of-the-art equipment scheduled inspection frequency model. PMID- 14564806 TI - The fundamentals of...electronic fetal monitoring. PMID- 14564805 TI - Accelerometer-based body-position sensing for ambulatory electrocardiographic monitoring. AB - Our objective is to validate the ability of 3 appropriately placed accelerometers to determine body position during ambulatory electrocardiographic (ECG) monitoring and to demonstrate the clinical applicability of this method. During ambulatory (Holter) monitoring, the ability to know a patient's position (lying down, sitting, standing, or changing from one position to another) is important in the evaluation of common symptoms such as dizziness, palpitations, and syncope. Changes in body position are also known to alter the electrical axis of the heart, resulting in artifactual changes in QRS amplitude and ST-segment morphology. We have developed an ambulatory patient-monitoring instrument that, through the use of microfabricated accelerometers, can simultaneously record body position information and 2 channels of ECG data. The accelerometers measure the effects of gravity and dynamic acceleration, allowing determination of a patient's orientation and movements. The accelerometer and ECG signals are input to a portable recorder and are filtered and digitized. Algorithms were developed to automatically determine body position. Ten healthy volunteers wore the device for 1 hour and followed a protocol of standing, sitting, walking, lying supine, and lying in the left and right lateral decubitus positions. An observer manually recorded times of position changes. Data were recorded and analyzed using software designed with MATLAB. The ability of the accelerometers and computer algorithms to determine body position was analyzed in terms of the sensitivity and specificity for each body position. The sensitivities for sitting, standing, walking, lying supine, lying right, and lying left were 98.8%, 99.2%, 95.5%, 99.1%, 98.9%, and 94.8%, respectively. The specificities were 99.7%, 99.4%, 99.6%, 99.0%, 99.8%, and 99.9%, respectively. The use of microfabricated accelerometers is a clinically feasible method to determine body position and can be applied to future studies correlating body position with ECG or other physiologic data. PMID- 14564807 TI - Simulation aids safety efforts. PMID- 14564809 TI - All screwed up. An obscure property of light puts a spin on astronomy. PMID- 14564808 TI - Creating a value-added risk management process. PMID- 14564810 TI - Why women work. PMID- 14564811 TI - Baffling the bots. Anti-spammers take on automatons posing as humans. PMID- 14564812 TI - The unseen genome: gems among the junk. PMID- 14564813 TI - The asteroid tugboat. PMID- 14564814 TI - An army of small robots. PMID- 14564815 TI - The future of string theory. PMID- 14564816 TI - Stranger in a new land. PMID- 14564817 TI - Flying on flexible wings. PMID- 14564818 TI - Why we sleep. PMID- 14564819 TI - Working knowledge. Nails and staples. PMID- 14564820 TI - Waiting for liftoff. PMID- 14564822 TI - Vibration induced nystagmus in normal subjects and in patients with dizziness. A videonystagmography study. AB - OBJECTIVE: To analyze the incidence and characteristics of vibration-induced nystagmus in normal subjects and in patients with vertigo. STUDY DESIGN AND SETTING: This is a prospective analysis of the effect of vibration on several points of the skull in 38 normal subjects, 10 patients who had undergone labyrinthectomy and 125 consecutive patients with dizziness referred to a tertiary care center. Nystagmus was documented and its corresponding slow phase velocity measured with a videonystagmography system. RESULTS: In 81.6% of normal subjects and 80.8% of all patients, nystagmus could be provoked by stimulation of at least one of the points on the skull to which the vibrator was applied. In normals, however, no subject had nystagmus greater than 2.8 degrees s-1 whereas 35.2% of patients did. CONCLUSION/SIGNIFICANCE: The value of the slow-phase velocity of vibration-induced nystagmus can be used to identify a sizeable proportion of patients with a vestibular disorders. Vibration-induced nystagmus is a frequent sign in patients with dizziness and can provide complimentary information about vestibular system function. PMID- 14564821 TI - [Nystagmus and vibration test research of mechanisms, theoretical methods: on 52 cases of unilateral vestibular lesions]. AB - OBJECTIVE: The aim of the present study was to examine the characteristics of the Vibratory Nystagmus (VN) optimise the topography and the frequency of the stimulus, determine the origin of the VN and analyse its clinical implications. MATERIAL AND METHOD: 52 severe unilateral vestibular lesions (SUVL) (post surgical vestibular areflexy) were studied. The vibratory nystagmus was measured by 2D and 3D videonystagmography (Synapsis, France). The stimulus was applied with a 3S vibrator at the vertex, both mastoids, and posterior cervical muscles at frequencies ranging from 20 to 150 Hz. For topographic optimisation, stimulation was given with S vibrator (Synapsis, France) at 100 Hz. The interferences between the vibratory test (VT) and the caloric test (CT) were studied on the normal ear (in 11 subjects). The same study was carried with the optokinetic test. RESULTS: The VT revealed a defective nystagmus at all frequencies of stimulation. Optimal response was obtained in a band frequency of 80-120 Hz. Stimulations at 100 Hz showed optimal responses for mastoid topography (responses in 96% of the cases). Stimulation at the posterior cervical muscles and at the vertex indicated respectively a response in 90% and 60% of the cases. The efficiency of the mastoid stimulation is not correlated with the side of stimulation (p = 0.9). The interference between the VT and caloric test (CT) at cold water (30 degrees et 20 degrees C) exhibited the inversion of the caloric nystagmus during the vibrator stimulation. The resulting nystagmus is respectively an algebric subtraction or addition between the pre-existing caloric nystagmus and the value of the vibratory nystagmus obtained before caloric test. Adaptation of the VN is moderate. The interference between the VT and the optokinetic test resulted in a subtraction or addition effect, according to the side of the lesion and the direction of the optokinetic stimulus. CONCLUSION: The VT is an efficient stimulation in mastoid topography. The vestibular contribution is bilateral by bony conduction of the vibration, it explores frequencies ranging from 30 to 120 Hz, with a maximum of response at 100 Hz. The VT interacts with the caloric test and the optokinetic test. The stimulation is very strong, and is able to inverse the caloric nystagmus at cold water stimulation (20 degrees C). In labyrinthine-defective subjects, the VN is always defective at all frequencies, whatever is the topographic location of the stimulus, and the position of the head. PMID- 14564823 TI - [Video-nystagmography and vibration test in the diagnosis of vestibular schwannoma. Review of 100 cases]. AB - OBJECTIVE: To evaluate informations given by the combination of videonystagmography (VNG) including vibratory tests and auditory brainstem responses (ABR) in patients suffering vestibular schwannoma (VS) and try to find the most conclusive test(s). Combination of different functional tests is supposed to improve diagnosis and preoperative evaluation and precise indication for magnetic resonance imaging (MRI) facing audiological and vestibular symptoms. MATERIAL AND METHOD: A prospective study of 100 patients with VS. All patients underwent a preoperative work-up including complete audiometry, auditory brainstem response (ABR) and videonystagmography (VNG). VNG protocol included caloric testing, rotatory tests, oculometry tests (saccade testing, optokinetic testing) and spontaneous and gaze-evoked nystagmus. From these six tests a score of positivity could be set, from 0 to 6. RESULTS: The vibratory test is non invasive and easy to realize. Were observed: 1/ a good sensitivity in vibratory test to elicit nystagmus in this context. 2/ a good correlation between subliminal rotatory chair tests and vibratory tests 3/ a better control of caloric testing using vibratory test. 4/ a good but deficient sensitivity of ABR alone with regard to VS (95%) 5/ an increase of sensitivity of VNG when coupling it with ABR and using as a criterion the score of positivity: no patient had all tests negative. CONCLUSION: The vibratory test is a non-invasive, fast examination with an easy execution. It reinforces VNG-ABR association screening power to diagnose VS. It constitutes, combined to caloric testing a good tool to diagnose and evaluate unilateral vestibular weakness. PMID- 14564824 TI - [Invasive cholesteatoma of the old radical mastoidectomy]. AB - OBJECTIVE: The aim of this study is to show the importance of computed tomography scanning (CT) and magnetic resonance imaging (MRI) in ancient radical cavities and to determinate the risk factors of an invasive cholesteatoma recurrency. MATERIAL AND METHOD: We present 4 cases of medial invasion in the petrous bone of a cholesteatoma without specific symptomatology which appeared many years after undergoing radical mastoidectomy for an acquired cholesteatoma. CONCLUSION: The analysis of this series allowed to put in evidence the factors of suspicion of an underlying cholesteatoma. They are: the age, an aural discharge, numerous surgical interventions, immunodeficiency, cholesteatoma in childhood and antecedent of facial nerve palsy. This trial emphasizes the importance of imaging (CT and MRI) in front of an ancient radical cavity which presents one or some of these factors in order to eliminate a recurrence of an invasive cholesteatoma. Let us note that the demand of the patients is sometimes aesthetic or functional. PMID- 14564825 TI - [Nasal polyposis: long term functional results in 203 patients treated by ethmoidectomy combined with intranasal corticotherapy]. AB - OBJECTIVE: Evaluate the long term quality of life in patients after ethmoidectomy associated with intranasal corticotherapy for nasal polyposis on the appreciation of the intensity of nasal symptoms. MATERIAL ET METHOD: The authors report their experience about 203 patients treated by endonasal endoscopic ethmoidectomy intranasal followed by a long term intransal corticotherapy, with a mean follow up of six years. In this prospective study, each symptom was evaluated using an analogic visual scale and a questionnaire. The evolution of asthma after surgery and the patients global satisfactory rate were noted. RESULTS: There is a global improvement of the nasal symptoms with a mean postoperative individual scores calculated at 32% for nasal obstruction, at 34% for rhinorrhea and at 51% for anosmia. No impact on asthma or improvement of asthma was encountered in 92% of the patients. The patients global satisfactory rate of this medico-surgical approach of the disease is 93.6%. A positive correlation was found between the severity of the olfactory disorders and the oral corticosteroid therapy. CONCLUSION: Endoscopic endonasal ethmoidectomy followed by intranasal corticotherapy represents a valuable protocol in treating patients with severe nasal polyposis. PMID- 14564826 TI - Electrorhinomanometric evaluation of nasal functions in transseptal transsphenoidal hypophysectomy. AB - OBJECTIVE: The prospective study in 23 patients, who were undergone transseptal transsphenoidal hypophysectomy, underlines the rhinologic aspects of this technique. MATERIALS AND METHODS: Preoperative and postoperative subjective complaints were compared, postoperative nasal respiratory functions were evaluated by means of electrohinomanometry. RESULTS: Nasal respiratory functions were preserved postoperatively in the patients with normal preoperative nasal anatomy (no statistical difference); while, in patients with obstructive deviation of the nasal septum, an improvement in nasal respiratory functions were noted as a result of simultaneously performed septoplasty (p < 0.01). CONCLUSION: Our results emphasize the importance of the collaboration of otorhinolaryngology and neurosurgery teams for the sake of postoperative nasal functions. PMID- 14564827 TI - [Surgery for hyperthyroidism on 43 patients]. AB - Treatment of hyperthyroidy uses surgery as well radioactive iodine and antithyroid agents. OBJECTIVES: The study analyses the modalities of hyperthyroidy surgery, describes its complications and the population undergoing surgical treatment. POPULATION AND METHODS: It is a retrospective study on 43 patients with hyperthyroidy treated by thyroidectomy from January first 1998 until 31 January 2002. RESULTS: The mean age is 48 years. The sex ratio showed a predominance of women (with 36 women and 7 men). The most frequent aetiology is Graves' disease and toxic multinodular goiter. Total thyroidectomy is applied to Graves' disease and toxic multinodular goiter while single toxic nodules are treated by loboisthmectomy. Surgery for hyperthyroidy have a low morbidity (1/43 haematoma, 0/43 hypoparathyoidy, 1/43 palsy of recurrent nerve). CONCLUSIONS: In this conditions, the hyperthyroidy surgery is fast, effective and does not need a heavy follow-up. Surgery seems to be a good alternative to antithyroid agents, which are constraining and often ineffective in the long term, and to radioactive iodine who leads to a long follow-up because of induced hypothyroidy. PMID- 14564828 TI - [Interest of the cervical gland dissection after radiotherapy with or without chemotherapy]. AB - OBJECTIVES: The aim of this study was to evaluate the necessity and morbidity of neck dissection after radiation therapy within organ preservation treatment. PATIENTS AND METHODS: We present a retrospective study of 64 patients, treated initially by radiotherapy with or without chemotherapy for squamous cell carcinoma of head and neck with cervical metastases (> 2 cm), who underwent post radiation neck dissection between January 1992 and August 2000. Eight (13%) patients were classified T1, nineteen (30%) T2, twenty (31%) T3, eleven (17%) T4 and six (9%) Tx. Eleven patients had N1 neck disease (17%), fifteen patients N2a (24%), eleven patients N2b (17%) and twenty-seven patients N3 (42%). RESULTS: Follow-up ranged from 3 to 86 months with a mean of 39 months. The average length of time between neck dissection and the end of treatment was 60 days. Complications were recorded in 21 patients (33%). Forty-four (68%) of 64 patients had microscopic residual disease. Eight (72%) of 11 patients with N1 neck disease and 17 (63%) of 27 patients with N3 neck disease had pathology. Initial N status was not a predictive factor of microscopic residual disease (p = 0.51). There was no significant relationship between clinical residual adenopathy and microscopic residual disease (p = 0.53). Fourteen patients are still alive without recurrent disease. Eight (57%) of these 14 patients had a positive pathology at the time of neck dissection. The mean follow-up time of these patients is 32.6 months, with a follow up longer than 2 years for half of them (n = 7). CONCLUSION: Neck dissection after radiation was planned for all patients with an initial node > 2 cm in diameter regardless of clinical response in the neck. We confirm that neck dissection appears to be safe after radiotherapy and is necessary because it improves quality of life and prevents fatal evolution with uncontrollable neck disease. PMID- 14564829 TI - Otoplasty for prominent ears in children. The technique adopted in the Portmann Institute. AB - The Otoplasty technique adopted in the Portmann Institute is a simple rapid technique that maintains the natural contours of the auricle with minimal risk of infection. The technique involves excision of a large ellipse of skin from the mastoid surface of the auricle and reflection of remaining skin to the edge of the helix. The subcutaneous and muscular tissues on the mastoid bone are excised and the mastoid surface of the auricular cartilage scored with monopolar diathermy. After haemostasis, the wound is closed using continuous long-term absorbable sutures. A dressing and bandage are applied and the child is monitored for 10 days. A head bandage is applied at night for one month with use of a sun screen cream on the scar at daytime. PMID- 14564831 TI - Auditory and phonetic perception of vowels in children with apraxic speech disorders. AB - The aim of this study was to assess auditory and phonetic perceptual processing of vowels in children with apraxic disorders, who demonstrated clinically with only a speech output deficit. Two experiments were conducted. In the preparatory Experiment 1 series of vowels were constructed by moving formant frequencies away from the extreme values in the vowel space in the direction of a 'neutral-vowel position'. These were presented to adults and children with no speech-language involvement. Based on identification performance low-redundancy vowels were selected, which served as the end-points of two vowel continua: /i/-/i/ and /a/ /a/. In Experiment 2 these continua were used in identification and discrimination tasks, presented to 11 children with apraxic speech problems (aged 6:11 to 9:6 years) and 12 normally developing children. The results showed poorer perception of vowels for the children with apraxic speech problems than for the control children for both continua. Identification functions indicated poorer phonetic processing; discrimination functions indicated poorer auditory processing. Furthermore, a combination of perception measures (identification and discrimination) proved to have a high differential and clinical value for the assessment of children with apraxic speech problems. The results support the view that subtle (subclinical) auditory processing deficits make part of speech output disorders. PMID- 14564832 TI - Lognormal distribution of pause length in ataxic dysarthria. AB - Previous investigations indicated that pauses in ataxic dysarthric speech are abnormal, but little is known about the nature of their variation or whether it can be characterized by a common distribution. The following types of pauses were measured in spontaneous monologues of 16 speakers with ataxic dysarthria: pauses that contained evidence of respiration, pauses that occurred between words, between phonemes, within phonemes, and those associated with stop consonants. Results showed that the duration of pauses not associated with stop consonants can be modeled with two distinct lognormal distributions. The lognormal distribution predicted duration of across types within 1.67% standard error, with the exception of pauses that occurred within phonemes. Distributional parameters pointed to some fundamental similarities between pauses that occur within words. The robustness of the lognormal distribution across pause types strongly supports this distribution as a viable and useful method of analysis. We hope this work will encourage others to consider the lognormal distribution. PMID- 14564830 TI - Toward an acoustic typology of motor speech disorders. AB - Acoustic methods have progressed to the point that an acoustic typology of the motor speech disorders can be constructed from a parameteric assessment of the speech subsystems (e.g., phonation, nasal resonance, vowel articulation, consonant articulation, intonation, and rhythm). The results of this analysis can be interpreted in respect to global functions in speech (e.g., voice quality, intelligibility, and prosody). This paper reviews studies showing that specific acoustic analyses have demonstrated or potential value toward the overall goal of constructing acoustic profiles of dysarthria and apraxia of speech. Several different acoustic measures are relevant to the study of the motor speech disorders, and these are increasingly supported by normative data and by guidelines for clinical application. Examples of these applications are discussed for a variety of specific neurologic diseases or perceptual types of disorder. Acoustic studies are useful in the study of motor speech disorders and recent progress points to a parametric analysis. PMID- 14564833 TI - The influence of sublexical and lexical representations on the processing of spoken words in English. AB - Previous research suggests that sublexical and lexical representations are involved in spoken word recognition. The current experiment examined when sublexical and lexical representations are used in the processing of real words in English. The same set of words varying in phonotactic probability/neighbourhood density was presented in three different versions of a same-different matching task: (1) mostly real words as filler items, (2) an equal number of words and nonsense words as filler items and (3) mostly nonsense words as filler items. The results showed that lexical representations were used in version 1 of the same-different matching task to process the words, whereas sublexical representations were used in version 3 of the same-different matching task to process the words. Finally, in version 2 of the same-different matching task individual variation was observed in the form of distinct sublexical and lexical biases. Implications for the processing of spoken words are discussed. PMID- 14564834 TI - [Measures enhancing the prevention of parasitic diseases in Russia]. PMID- 14564835 TI - [In vitro induced activity of polymorphonuclear leukocyte myeloperoxidase at different stages of opisthorchis invasion and at stages of a different rehabilitative period]. AB - Polymorphonuclear leukocytes from 115 patients with opisthrochiasis at its different (acute and chronic) stages were studied before and 1-3 days, 1-3 months, and 1-2 years after antihelminthic therapy with bilthricide. The functional activity of the cells in question was evaluated following an hour load with different drugs: bilthricide, ascorbic acid, decaris, cyclophosphane, doxycycline in a final concentration of 2.5 x 10(-5) mole/l. Physiological solution and autologous plasma were test controls. The changes in the activity of extracellular (secreted), intracellular (reserve), and summary myeloperoxidase on incubation with each test agent were found to depend on the stage of the disease and the interval after antihelminthic therapy. In early Opisthorchis invasion, the maximum increase in the induced activity of secreted myeloperoxidase is followed by a fall in the level of reserve myeloperoxidase in response to these drugs. With this, there is the most pronounced decrease in reserve myeloperoxidase of polymorphonuclear leukocytes in chronic opisthorchiasis, which may be due to the disrupted controlling processes of its production at late stages of the disease. The distinctive features of the action of agents on the overall activity of granular leukocytic myeloperoxidase in acute and chronic opisthorchiasis suggests that autologous plasma, ascorbic acid, and doxycycline should be used as a test control at the clinical stage of Opisthorchis invasion. PMID- 14564836 TI - [Ecology and the genetic structure of sympatric leishmania species circulating in the intra-continental deserts of the south Palaearctic region]. AB - A PCR fingerprinting approach with single non-specific primers--(GTG)5 and T3B- was apply to investigate variations in the genotyping of three species of Leishmania species within the Rhombomys opimus area. Forty-three strains of Leishmania major, L. turanica, and L. gerbilli circulating among great gerbils in Turkmenistan, Uzbekistan, Kazakhstan, and Mongolia were examined. PCR fingerprint revealed a high genetic intraspecific heterogeneity among L. turanica strains. Three groups of strains were clearly identified. The strains from Mongolia greatly differed from other L. turanica ones. The second group was formed by strains from Kazakhstan, they also demonstrated rather different patterns. L. turanica strains from Turkmenistan and Uzbekistan showed only minor differences, but greatly different from those from Kazakhstan and Mongolia. The groups identified by the PCR fingerprint correlate with the conditions of circulation: the duration of a transmission season and as the result of different periods of retention of Leishmania in the skin of great gerbils, as well as the presence or absence of L. major as coexisting species. There were no differences between L. turanica strains isolated from different hosts in the same geographical region, as well as between L. turanica strains isolated in the hyper- or meso and hypoendemic foci. There was no correlation between the genotypic and phenotypic characteristics of L. turanica. No intraspecific polymorphism was found among L. major and L. gerbilli strains from different geographical regions within the great gerbil area. PMID- 14564837 TI - [Indication of Borrelia burgdorferi sensu lato in the taiga tick larvae obtained from spontaneously infected females]. AB - Hungry larvae from 17 clutches by spontaneously infected Ixodes persulcatus females were examined by inoculation of the BSK II medium, by microscopy of fixed smears, and by polymerase chain reaction (PCR) with the primers specific toward the conserved sequences of Borrelia burgdorferi 16S ribosomal RNA gene. A study of 781 larvae individually or as pools could not reveal Borrelia DNA by PCR. Inoculations of the BSK II medium with 600 larvae yielded no positive results either; immobile spiral forms of Bacillus spp. were detected in 16 (26.7 +/- 5.7%) cultures of the larvae obtained from 7 females. Microscopy of 1416 fixed smears showed typical Borrelia in 7 (0.5 +/- 0.4%) specimens of larvae resulting from the same hatch; spirochaete-like cells were present in 13 (0.9 +/- 0.5%) cases. The single female from those of the 17 hatches transmitted Borrelia through eggs to filial-generation larvae. The infection rate of the larvae emerged from the eggs of this hatch was 7% and their individual infection rates were 0.4-0.8 Borrelia per 100 fields of vision. The findings suggest that there may a transovarian transmission of Borrelia in I. persulcatus ticks, but its likelihood is very little. PMID- 14564838 TI - [Genotyping of spotted tick fever rickettsiae, detected in Russia and Kazakhstan]. AB - R. slovaca was first detected in the ticks D. marginatus gathered in the Stavropol Territory and the Voronezh Region (European Russia). The recently discovered rickettsial genotype DnS14 was first found in the ticks D. silvarum from Buryatia and D. niveus from the Karaganda Region (Central Kazakhstan). The rickettsial genotype RpA4 was most common in the ticks of the genus Dermacentor in Russia and Central Kazakhstan. An analysis of the spread of rickettsias of the STF group shows their close ecological relation to definite types of Ixodes. The rickettsias R. slovaca and RpA4 co-exist in the ticks D. marginatus and D. reticulatus (the western part of a Dermacentor area in Eurasia) and DnS14 and R. sibirica do in D. nuttalli and D. silvarum (the eastern part of the area). D. marginatus and D. reticulatus in the areas characterized by the most specific saturation of a Dermacentor area (the south of West Siberia) are carriers and reservoir of R. sibirica. The rickettsial genotype DnS28 may be now considered to be environmentally associated with one species of ticks--D. nuttalli. At least 6 genotypes of STF rickettsias--R. sibirica, R. astrahan fever (R. conorii), R. slovaca, RpA4, DnS14, DnS28--has been currently identified in Russia and Kazakhstan. PMID- 14564839 TI - [Influence of a temperature factor on the vector capacity of the flea Citellophilus tesquorum altaicus]. AB - The impact of temperatures (2-3 to 23-25 degrees C) of flea keeping on their vector capacity was studied in the experiment. In hibernated fleas, the relationship of their vector capacity to the temperature of their keeping, estimated by the level of plague block in species and by the case rates of animals was insignificant, i.e. there is a trend for these indices to increase with elevated temperatures. In the fleas of current years breeding, the rate of block significantly increased with a rise of temperatures from 5-6 to 23-25 degrees C. There were also increases in the number of transmissions of a plague pathogen to susliks and in the number of animals with generalized infection with a rise in temperature up to 16-18 degrees C. The findings and constructed regression equations may predict the pattern of changes in the vector capacity of the flea C. tesquorum altaicus in the study range of temperatures in the enzootic area of a Tuva natural focus of plague. PMID- 14564840 TI - [Comparative study of ixodes ticks on their infection with tularemia pathogen using biologic and serological assays]. AB - The paper presents the results of a long-term comparative study of the infection of Ixodes tick imagoes with the pathogen of tularemia by using the biological assay and antibody neutralization test. The infection rates determined by both methods were found to be identical or close. If there was a difference in the rates in some years, it was not statistically significant. A statistically significant difference could not be found in the infection rates for ticks and in the dependence on their species composition though the level of infection in representatives of one species greatly differed from that in others. The serological study of Ixodes tick imagoes, aimed at determining their infection with a tularemia pathogen, yielded the results which are comparable with those of the biological assay and may be used in epizootological studies without using the biological test or by significantly limiting its use. PMID- 14564842 TI - [Long-term morbidity and spatial distribution of tick-borne encephalitis and ixodes tick-borne borreliosis in Udmurtia]. AB - The paper considers the trends of morbidity due to tick-borne encephalities (TBE) and lxodes tick-borne borrelioses (lTBB) in Udmurtia during 35 years (1965-2000) and evaluates the long-term influence of measures aimed at suppressing natural foci by eliminating the basic vector on this process. For this, data on 1062 cases of fevers occurring after the bite of a tick in Udmurtia in 1965-1968, the similar data on 1509 cases in 1983-1987, and the official data on the incidence of these infections in 1996-2000 have been retrospectively analyzed. It has been shown that in Udmurtia the natural foci of lTBB coexisted with those of TBE long before the pathogens of borrelioses were described and notification of the infection of this group was initiated. In the 1990s, there was a drastic rise in the incidence of TBE and lTBB as compared with the 1960s and 1980s, which was due to an increase in the rate of contacts of the population with natural foci and to the better diagnosis of these infections. The prevalence of lTBB coincides with that of TBE. There is a long (more than 30 years) epidemiological effect in eliminating the vector Ixodes persulcatus. PMID- 14564841 TI - [Spontaneous infection of ixodes ticks with salmonella]. AB - An epizootological study of the ticks gathered from the cattle and ground surface in the Dedoplistskaroisky and Dushetsky districts of Eastern Georgia has identified 5 Salmonella a typhimurium strains. The strains were isolated from the ticks Haemaphysalis punctata, Rhipicephalus bursa, Hyalomma plumbeum. They were found to be resistant to 10 of 16 antibiotics. The strains showed a high sensitivity only to 3 antibiotics, such as gentamicin, cirpofloxacin, and nitrofurantoin and 3 strains were highly sensitive to chloramphenicol. The above ticks seem to have been spontaneously infected with the pathogen Salmonella on the animals at the moment of bacteremia. PMID- 14564843 TI - [Changes in activity of natural foci of plague in Tuva region]. PMID- 14564845 TI - [Stages of laboratory and field evaluation of the insecticidal activity of active substances and preparative forms]. AB - The paper describes the basic stages of study of the insecticidal activity of compounds used for medical dosinsectation. The classes of efficacy of test substances are characterized. PMID- 14564844 TI - [The new agent G-1724: synthesis and test on murine models of trichuriasis and trichinosis]. AB - To search for new antihelminthics, the new compound N-[3-chloro-4-[(1-ethyl-2 benzimidazolyl) thio]phenyl]-2-hydroxy-3,5-dibromobenzamide (the agent G-1724) was synthesized and tested by using the models of trichocephaliasis (Trichocephalus muris) and trichinosis (Trichinella spiralis). The tests demonstrated that this agent had an nematocidal effect (71-78%). PMID- 14564846 TI - [Human and animal echinococcosis in the Stavropol territory]. PMID- 14564847 TI - [Leptospirosis outbreaks in the Ul'ianovsk region]. AB - The paper presents the data of an epidemiological investigation on the outbreak incidence of leptospirosis in the Ulyanovsk Region. One hundred and forty-one recording forms under No. 171/y filled in for patients with leptospirosis have been examined. All the outbreaks have been found to be of seasonal nature (June to August) and to show an aqueous transmission of infection. The occupational nature of the disease was absent among the patients. PMID- 14564848 TI - [Human dirofilariasis in the Moscow region]. AB - Dirofilaria repens Railliet et Henry, 1911 was for the first time studied in the Moscow Region where 11 patients (6 males and 5 females) aged 4-72 years were recorded in 2000-2002. The geographical distribution (the southern and south eastern parts of the region) of local cases of dirofiliriasis, was observed. There was a relatively high rate of D. repens in the male genitalia of the infected patients as compared with that reported by other authors. In accordance with the current terminology, D. repens in the Moscow Region is proposed to be regarded as emerging infection. The authors show it necessary to obligatorily record all autochtonous cases of D. repens infection, to make epidemiological surveillance and analysis in order to early detect and prevent dirofilariasis associated complications. PMID- 14564849 TI - [Animal and human cestoda infection (sparganosis)]. AB - Spirometrosis (sparganosis) in animals and man is a serious ecological problem in Russia, which requires the constant attention of veterinary and medical services. The paper presents data on the biology of the parasite Spirometra erinacei europaei Muller, 1937, an account and diagram of a developmental cycle, and original photos of parasitic eggs, plerocercoids, strobiles, and cestode joint from animals. PMID- 14564850 TI - Healthy shiftwork, healthy shiftworks. AB - Reflecting diversifying shift systems, extensive effort is put into managing shiftwork and reducing safety and health risks. It is accepted that shiftworkers are exposed to particular risks inherent in their irregular work schedules. This raises the question of how and to what extent we can ensure healthy work life for shiftworkers. In answering the question, we need to identify effective measures to improve both shiftworking conditions and the health of shiftworkers. Based on recent experiences in managing shiftwork, we note three directions of such measures: (a) comprehensive action to avoid risk-enhancing conditions based on general guidelines, (b) risk control as to workload, worksite ergonomics and risk reduction, and (c) support for flexible and restful working life. International standards are obviously relevant to these three aspects. Our own experiences in applying a set of ergonomic checkpoints to plant maintenance shiftwork demonstrate the usefulness of focusing on flexible work schedules and on multiple job-related factors such as night workload, ergonomic environment, resting conditions and training. There is a strong need for participatory planning and implementation of multi-area improvements as well as for relying on flexible schedules and autonomic teamwork. We may conclude that healthy shiftwork and healthy shiftworkers are compatible with each other only when certain conditions are met. In achieving this end, we need to combine (a) comprehensive measures to improve work schedules and job life, (b) strict risk management and (c) locally adjusted participatory steps for continual improvement. PMID- 14564851 TI - Strategies for the implementation of new shift systems. AB - Implementation barriers may be caused by deficiencies in the knowledge, skills, motivation, or support of those involved in the process of implementation of a new shift system. There is no 'one and only' way of planning and implementing a new shift system. However, if the following factors of success are taken into consideration there is a better chance that workers will accept a new shift system: worker participation, information, communication, training, promoter commitment, professional project management, tailor-made solutions and an adequate organizational framework. These factors are particularly relevant in addressing barriers to the implementation of new shift systems. The most important measures to cope with resistance to change of shift systems are: worker participation, information, communication, training, promoter commitment, professional project management, tailor-made solutions and an adequate organizational framework. PMID- 14564852 TI - The 24-hour society between myth and reality. AB - The 24-hour society appears to be an ineluctable process towards a social organisation where time constraints are no more "restricting" the human life. But, what kind of 24-hour society do we need? At what costs? Are they acceptable/sustainable? Shift work, night work, irregular and flexible working hours, together with new technologies, are the milestone of this epochal passage, of which shift workers are builders and victims at the same time. The borders between working and social times are no more fixed and rigidly determined: not only the link between work place and working hours is broken, but also the value of working time changes according to the different economic/productive/social effects it can make. What are the advantages and disadvantages for the individual, the companies, and the society? What is the cost/benefit ratio in terms of physical health; psychological well-being, family and social life? The research on irregular working hours and health shows us what can be the negative consequences of non-human-centered working times organisations. Coping properly with this process means avoiding a passive acceptance of it with consequent maladjustments at both individual and social level, but adopting effective preventive and compensative strategies aimed at building a more sustainable society, at acceptable costs and with the highest possible benefits. PMID- 14564853 TI - Interactive computer aided shift scheduling. AB - This paper starts with a discussion of computer aided shift scheduling. After a brief review of earlier approaches, two conceptualizations of this field are introduced: First, shift scheduling as a field that ranges from extremely stable rosters at one pole to rather market-like approaches on the other pole. Unfortunately, already small alterations of a scheduling problem (e.g., the number of groups, the number of shifts) may call for rather different approaches and tools. Second, their environment shapes scheduling problems and scheduling has to be done within idiosyncratic organizational settings. This calls for the amalgamation of scheduling with other tasks (e.g., accounting) and for reflections whether better solutions might become possible by changes in the problem definition (e.g., other service levels, organizational changes). Therefore shift scheduling should be understood as a highly connected problem. Building upon these two conceptualizations, a few examples of software that ease scheduling in some areas of this field are given and future research questions are outlined. PMID- 14564854 TI - Flexible work hours, health and well-being in the European Union: preliminary data from a SALTSA project. AB - Demand for flexible work hours (FWH) is increasing in Europe aimed at increasing the number of production hours on one hand, and, on the other, reducing individual working hours and/or increasing autonomy and control on them. In view of the lack of knowledge of the effects of FWH on health and safety, we started a pilot project, funded by the Joint Programme for Working Life Research in Europe (SALTSA), aimed at: a) comparing the most relevant national legislation and how the EU Directive 93/104 "concerning certain aspects of working time" has been implemented in the member States; b) reporting prevalence and trend of FWH in Europe according to the three EU Surveys on Working Conditions carried out in the last decade; c) collecting practical examples of innovative FWH; d) evaluating their impact on health and safety in relation to work sectors, job demands, social life, aging and gender. Consequent actions are going to include information and consultancy for pertinent authorities and social parties involved, as well as training programmes for Union officials and similar groups concerning the organisation of FWH according to ergonomic principles. PMID- 14564855 TI - Flexible working hours and well-being in Finland. AB - Flexibility of working hours became more prevalent in the 1990s in Finland. According to a representative survey on Finnish wage and salary earners (n = 1790) at the beginning of 2000, a great majority of male (76%) and female (65%) employees regularly worked overtime and/or had irregular working hours every month. These employees were flexible in meeting the needs of their companies/employers. Individual flexibility of working hours was far less common, only one third of male and female employees were able to regulate their working hours. A better balance between company-controlled and individual flexibility would, however, improve the well-being of employees. Employees working overtime without being allowed to regulate their working hours felt more symptoms of distress and had more conflicts in combining workplace and family roles than those who could individually determine their working hours flexibly. An investment in individually determined flexibility, for example by means of participatory planning, would improve the well-being of employees, and thus also improve the productivity of the organization. PMID- 14564856 TI - The French 35-hour workweek: a wide-ranging social change. AB - The reduction of the legal working week to 35 hours in France has generated wide ranging social change. We examine the resulting changes in working-time patterns as well as their repercussions on the use of the time gained and on the quality of life and health. To compensate the reduction in the length of the working week, companies have modified the working-time patterns, by extending operation time (shiftwork, atypical schedules) and by matching the on-site workforce to production requirements (flexible working hours). They have sought to make more efficient use of working time: job intensification or job compression. The effects on the off-the-job life and health are linked to the shiftwork and atypical schedules designed to increase the company's operating time, and adjustments to the company's need for flexibilization impose working time/free time patterns that are at odds with biological rhythms and social life patterns. Changes to working-time patterns have unexpected consequences for work organization: heightened difficulties for the individual and the crew. These changes may generate a range of health problems related to overwork and stress. The way some companies have adapted may call into question the usefulness of work done by employees, thus damaging their social identity and mental well-being. PMID- 14564857 TI - Flexible working time systems--the process of design and implementation. AB - Examples of the design and implementation processes of flexible working time systems are derived from a counselling project in North Rhine-Westphalia, Germany. The practical cases shown represent a diversity of variable or flexible working time systems according to different branches and different anticipated aims to reach with flexibility. For companies flexibility should improve or at least consolidate their economic situation, and for employees flexibility should result in a beneficial effect on health, family and social life. Three examples of counselling practice are reported in this paper, each case representing a different approach to flexibility in working time. To avoid overtime an industrial production plant expanded the weekly operating time without increasing the employees working time. For this purpose shifts off had to be planned. To provide a continuous service one department of a service company changed from duty on call to regular shifts on weekends. Responsibility in handling the new working times was assigned to the team. In an old people's home working times were designed according to the irregular demands throughout the day, in combining full and part-time jobs adequately. PMID- 14564858 TI - The effects of double-shifts (15.5 hours) on sleep, fatigue and health. AB - The aim of the present study was to investigate how "double-shifts" (15.5 hours) affects sleep, fatigue and self-rated health. The study was carried out on male construction workers of which 80% were long-distance commuters. The schedule involved two work periods and each work period involved two double shifts in a row. The subjects filled in a sleep/wake diary at 8 times across a year and a questionnaire at 3 times. They also wore an actigraph during one shift cycle. The results showed that sleepiness, and to a certain extent, mental fatigue increased during double shifts and accumulated across days. The short rest time (8.5 hours) between days caused insufficient sleep and approximately 5.5 hours of sleep was obtained between double shifts. Questionnaire data showed that complaints of insufficient sleep, exhaustion on awakening and pain symptoms increased across the year. It was concluded that a shift system involving double shifts has a negative effect on fatigue, recovery and health-related well-being. PMID- 14564859 TI - Predicting shiftwork-related outcomes: shiftwork locus of control and circadian type. AB - This research discusses the use and viability of the shiftwork locus of control construct alongside circadian type measures as a potential predictor of shiftwork related outcomes. The shiftwork locus of control (SHLOC) scale, measures of circadian type and shiftwork-related outcome measures were completed by 100 shiftworkers on two occasions separated by seven months. The SHLOC scale measures shiftworkers' generalised beliefs about the levels of personal control they perceive in relation to four major functional domains commonly associated with shiftwork-related disruption. These domains include: sleep, social, health and work problems. The results of multivariate regression analysis showed the SHLOC scale to be predictive of the experience of shiftwork-related sleep and social life problems while the circadian type measures were predictive of alertness at 7 months. The results suggest that a constellation of personality factors may be an important influence on an individual's tolerance to shiftwork. PMID- 14564861 TI - A study of nurse scheduling in Japan. AB - Scheduling nurses to staff shifts is a major problem in hospitals. The necessity of maintaining a certain level of service and skill in the makeup of every shift, while balancing the workload among the nurses involved, is incredibly difficult. It is often impossible to develop a schedule which satisfies all the requirements despite the time and resources spent in the effort. This paper summarizes all our published research on nurse scheduling to date. The difficulties realized by our two investigations in Japan are shown first, together with a resulting scheduling problem. The nurse scheduling model based on the results is then described. In this model, all constraints are divided into two essentially different types; that which maintains a certain level of skill for each shift ('shift constraints') and that which concerns the workload for each nurse ('nurse constraints'). By classifying the constraints in this manner, we can determine what is affected by a specific constraint when the constraint is not satisfied. We developed efficient algorithms while taking advantage of the structure of this model. Finally, it is shown that our algorithm can solve this problem for a 2 shift system efficiently. PMID- 14564860 TI - The effects of a roster schedule change from 8- to 12-hour shifts on health and safety in a mining operation. AB - The current study examined the impact on employee health and safety of changes to the roster system of an Australian coal mine. Absenteeism and incident frequency rate data were collected over a 33-month period that covered three different roster schedules, an 8-hour system, a 12-hour system and a 12-hour system incorporating unregulated overtime. The first change was implemented after consultation with the employee population, whereas the second was not. There were no significant negative effects of the 12-hour pattern, when compared to the 8 hour system. However, when unregulated and excessive overtime was introduced as part of the second round of changes, absenteeism rates were increased in one sector of the mine. The maintenance sector was subject to a significant increase in absenteeism rates, which may have been attributable to the excessive overtime required of the workers in that area. It is important that overtime be strictly monitored and that the employee population are involved in the process of roster change. PMID- 14564862 TI - Balancing flexibility for the employer and the employee: a case study of the development of annualized hours employment contracts. AB - As with many forms of flexible working, Annualized Hours (AH) systems offer potential benefits to both the employer and the employee. However, the flexibility requirements of employers and employees often conflict. Therefore, when a large food manufacturing organization decided to redesign its AH system, it employed an independent consultancy to act as neutral third party. The consultancy provided technical expertise and assistance in developing an AH system that optimised productivity and was acceptable to the workforce. Data are presented, obtained from focus groups conducted throughout the organization, describing some of the potential difficulties of implementing an AH system. Drawing upon these data, a number of new AH systems were proposed and modelled using specialist software tools. The design process is described, together with the advantages and difficulties associated with use of the software tools. It is concluded that the key elements in the process of designing AH systems are centred around issues of trust and communication; the involvement of a broad range of interested parties, through a process of carefully managed group facilitation; and the need for adequate technical support in the development and evaluation of AH systems. PMID- 14564863 TI - Reducing night shift exposure: a pilot study of rota, night shift and age effects on sleepiness and fatigue. AB - The reduction of exposure to longer runs of successive night duties is often one of the aims of 'best practice' intervention. The objective of this pilot study was to examine the effects of a change in rota upon sleepiness and fatigue, and potential age-related issues in relation to the change. Participants were 102 police officers, mean age 33.60 (sd 6.11), mean shiftwork experience = 10.56 years (sd 6.84). Testing occurred at least one month before the change from the existing 'Ottawa' system to a new rota that split a run of 7 consecutive night shifts into blocks of 3 and 4 within the rota cycle, and 6 months after the change. Measures of critical flicker fusion, subjective fatigue and sleepiness, and stimulant intake were taken in relation to selected night shifts on both rotas. Older officers tended to report higher fatigue and sleepiness, and higher caffeine intake than the younger shiftworkers irrespective of rota or shift. The findings tend to suggest that the change of rota had little effect but that age may play a substantial role in responses to night work. PMID- 14564864 TI - Can we predict perceived risk? AB - This paper examines the possibility that we may be able to use subjective measures of perceived risk in order to assess the relative safety of different shift systems. A large-scale survey of safety-critical engineers included three items relating to risk on each shift, namely alertness, likelihood of making a mistake and confidence in driving home after it. These three measures were found to load on a separate factor for each shift. Hierarchical regression analyses indicated that perceived night shift risk could be predicted on the basis of circadian type, the extent to which the engineers could control their work schedule, and a number of features of the scheduled shift system. In most cases the relationships observed were reasonably consistent with established trends in either performance capability or accident and injury frequency. However, there were exceptions to this indicating that results based on measures of perceived risk should be interpreted with the utmost caution. PMID- 14564865 TI - Time on task effects on safety. AB - Reviewing the literature on time on task effects on safety shows contradictory evidence, especially with regard to 12 h shifts. It is argued that this might depend on methodological problems associated with the analysis of accident data, e.g. selectivity of samples, validity of data bases and study designs, especially for analyses at the company level. Analyses of aggregated data seem to indicate an exponential increase of accident risk with time on task beyond the normal working day. This is supported by some recent studies based on data from the Federal Republic of Germany. PMID- 14564866 TI - Prevention of accidents by transportation shiftworkers. AB - For commercial transportation operations, around-the-clock operations are not a recent development. Despite the tenure of these practices, many accidents related to hours worked continue to be a problem. Although efforts by shiftwork experts to solve these problems are often mode specific, the transportation field does suggest several alternative approaches, which may be relevant to general shiftwork problems outside the transportation area. Four contemporary approaches to transportation shiftwork are discussed: environmental design, equipment design, organizational design, and information access systems. Examples are presented within each of these approaches. PMID- 14564867 TI - Legal issues in accidents caused by sleepiness. AB - The impact of shift-work on sleep, performance and general health appears to be substantial. The most immediate consequence of night shiftwork is sleep loss. The aim of the present paper is to describe legal cases involving accidents attributed to sleepiness or fatigue, mainly as a consequence of shift-work or prolonged work hours, in the UK, USA and Australia. The paper will describe how legal systems are dealing with such incidents and how this may change in the future. Accidents related to sleepiness may result in criminal prosecution, for example charges of culpable driving. For acts performed while a person is sleeping (e.g., motor vehicle accidents), the legal question of voluntariness may be raised. The issue of employers' liability in such cases is contentious. Special liability regimes are in place to cover employers' liability. Employers may be deemed liable for injuries of third parties caused by wrongful acts of employees committed in the course of their employment. In the future, it is likely that employers will need to take greater precautions to reduce sleepiness and fatigue in the workplace, especially where the risk to public and environmental safety, health and productivity are significant. PMID- 14564868 TI - Health and safety among film technicians working extended shifts. AB - This paper presents selected results from a pilot research. The study of film technicians' work schedules and occupational hazards was based on a questionnaire administered to all 2200 film technicians (650 valid replies; response rate = 30%), interviews with producers and technicians on film-shooting organization and scheduling, courses in film technique, individual interviews with workers in trade at high risk, and review of the literature on freelancers, on the effects of intermittent work, and on risk factors for musculoskeletal injuries. Work schedules showed a tendency to extremely long work shifts (14 hours per day on average, and up to 19 hours in some trades). Occupational constraints and work schedules were found to relate to an increased risk of work-related injuries. Technicians identified fatigue associated with work schedules as the principal risk of accidents and one of the factors responsible for causing or aggravating their many musculoskeletal injuries. Work schedules were not the only cause of these injuries: stress--due to time constraints, work responsibilities and job insecurity--was also an important risk factor, consistent with the literature on musculoskeletal disorders. Physical workload was also problematic, particularly when demanding tasks had to be performed under severe time constraints. PMID- 14564870 TI - Comparing the effects of fatigue and alcohol consumption on locomotive engineers' performance in a rail simulator. AB - Laboratory studies have established that the performance impairments due to fatigue and alcohol consumption are quantitatively similar. However, the generalisability of this phenomenon is not clear because comparisons have not been made in realistic work settings with experienced shiftworkers. The aim of the current study was to quantify the effects of fatigue on performance in a simulated work environment (i.e. rail simulator) and compare them with the effects of alcohol consumption. It was hypothesised that fatigue would significantly impair driving performance, and that this impairment would be quantitatively similar to that associated with moderate levels of alcohol consumption. Twenty locomotive engineers participated in the study with a randomised cross-over design and three conditions: baseline, fatigue, and alcohol. During each 8-hour condition, participants completed four driving sessions in the rail simulator. The results indicate that fatigue caused participants to disengage from operating the simulator such that safety was traded off, not necessarily deliberately, against efficiency. The impairment in safety due to fatigue was in a range similar to the impairment associated with moderate levels of alcohol consumption. In summary, the study demonstrated that the effects of fatigue in a simulated work environment can be quantified and may be considerable. PMID- 14564869 TI - Beneficial effects on employment and preventive health care: the concept and results of counselling processes from a German project. AB - Since December 1998 a project of working time counselling named "Design of Modern Working Times" is supported by the federal state of North Rhine-Westphalia in Germany and the European Union; AWiS-consult is entrusted to carry out this project. The service offered to all companies, institutions, work committees, and employees interested covers information, assessment of existing working time schedules, development of alternative or new working time schedules, and accompanying implementation processes. Up to June 2001 nearly 300 inquiries were worked on, most of them from industry, service companies, and from the field of care and hospitals. Working time design in this project aims not only at work safety and health care but also at beneficial effects on employment. In a special campaign from May 2000 to May 2001, 61 companies were supported by AWiS-consult, with the result of more than 400 new employees and 16 companies with employment secured, so the combination of preventive health care and beneficial effects on employment by working time design can be assessed as successful. PMID- 14564871 TI - The effect of shiftwork on driving to and from work. AB - A questionnaire study was conducted to obtain information from shiftworkers on the levels of tiredness experienced on the drive to and from work at different times of the day. The results demonstrated that, compared to non-shiftworkers, shiftworkers were more tired on the drive between their home and the workplace and more at risk of falling asleep behind the wheel. The main factors contributing to this increased level of sleepiness and associated driving impairment were the length of the sleep period prior to work, the type of shift and the travelling time. There was some evidence to suggest that on working days shiftworkers were more likely to be involved in an accident than non shiftworkers. However, there were insufficient data to determine whether accidents were correlated with driver sleepiness. PMID- 14564872 TI - Performance, sleep and circadian phase during a week of simulated night work. AB - The current study investigated changes in night-time performance, daytime sleep, and circadian phase during a week of simulated shift work. Fifteen young subjects participated in an adaptation and baseline night sleep, directly followed by seven night shifts. Subjects slept from approximately 0800 hr until they naturally awoke. Polysomnographic data was collected for each sleep period. Saliva samples were collected at half hourly intervals, from 2000 hr to bedtime. Each night, performance was tested at hourly intervals. Analysis indicated that there was a significant increase in mean performance across the week. In general, sleep was not negatively affected. Rather, sleep quality appeared to improve across the week. However, total sleep time (TST) for each day sleep was slightly reduced from baseline, resulting in a small cumulative sleep debt of 3.53 (SD = 5.62) hours. Finally, the melatonin profile shifted across the week, resulting in a mean phase delay of 5.5 hours. These findings indicate that when sleep loss is minimized and a circadian phase shift occurs, adaptation of performance can occur during several consecutive night shifts. PMID- 14564873 TI - Sleepiness and recovery in schedule change and the eighty-four hour workweek. AB - The aims were to evaluate sleepiness and recovery during a schedule change, and during an 84-hours workweek. The control group (16 men) stayed on a six-week schedule, whereas the intervention group (12 men) transferred to a seven-week schedule. Sleepiness was estimated, using the KSS-scale, four times during the first and the third night in the fifth or sixth shift week. Recovery was assessed through four estimations on days one, three and five during the week off. Statistical testing was carried out using repeated measurement ANOVA. Sleepiness at night was affected by night (F = 4.90, p < 0.05) and hour (F = 33.64, p < 0.001) in both groups. The intervention group was sleepier during the first recovery day compared to the control group (F = 4.02, p < 0.05). Analysis of the 84-hour-week showed an effect of night (F = 8.98, p < 0.05) and hour (F = 71.60, p < 0.001) on night work, and day (F = 22.49, p < 0.01) and hour (F = 6.66, p < 0.05) on recovery. Sleepiness was more pronounced on the first recovery day (F = 23.08, p < 0.01). The seven-week schedule showed no effect that differed from that of the control group on sleepiness during the night shift. After the 84-hour workweek the workers recovered in about three days. The new schedules may affect the first recovery day negatively. PMID- 14564874 TI - Cross-sectional survey on risk factors for insomnia in Japanese female hospital nurses working rapidly rotating shift systems. AB - A cross-sectional survey was carried out for the purpose of examining the association of work stress with insomnia among nurses working rotating shift systems. A self-administered questionnaire on sleep and mental health was distributed to 875 nurses at five hospitals in Japan. Out of the data from 785 (89.7%) respondents, those from 555 women engaged in rapidly and irregularly rotating shift systems as full-time nurses were examined. The prevalence of insomnia (29.2%) was three to four times higher than that in the general population. Among the insomniacs, 23% were current users of medically prescribed hypnotics. Multiple logistic analysis revealed that being 24 or less years old, working three or less night shifts per month, having six or less non-working days per month, receiving less support from colleagues and superiors, and taking care of severely ill patients were independent risk factors for insomnia. The number of hours of a night shift was found not associated with insomnia, taking the above variables into account. These results suggest that at least three factors, i.e., 1) young nurses not adapted to shiftwork, 2) the nurses requiring to remedy sleep deficits on non-working days, and 3) the work stress partly characteristic of nurses, have an adverse influence on their sleep. PMID- 14564875 TI - Shiftworkers in developing countries: health and well-being and supporting measures. AB - Developing countries currently represent approximately 79% of the world population. Living and working conditions in middle and low income countries are harder and worse than for workers in industrialized countries. In developing countries, workers usually face more dangerous workplaces and unhealthy environmental and occupational conditions than those of industrial countries. Shiftworkers can face even more difficulties, due to constraints caused by their working time and consequences on health. Occupational health actions oriented by health policies were implemented during the 1990s in several Asian countries and in Brazil. these actions are important to promote workers' health. The general aims of this report are: a) to discuss topics related to equity and health; b) to present the main items of international and Brazilian legislation for shiftworkers; and c) to review general and specific measures of occupational health for shiftworkers in developing countries. In order to have equity on health and well-being, supporting measures should encompass micro and macro improvements at local, regional and national levels. Governmental and non governmental organizations, professional bodies, labor unions, research institutes, universities, technical schools and syndicates, would play important roles to achieve these goals. PMID- 14564876 TI - Shiftwork in the hot environment. AB - The study examined the risks of heat induced workplace accidents (textile industry, N = 4125) and the heat tolerability of the rotating day (morning and afternoon, N = 16) and permanent night shift workers (N = 13) in hot-dry and hot humid environment (34 to 49 degrees C, 50-80% RH; 31 to 42 degrees C ET, Basic). Accident prevalence was significantly high in the summer months (May-June) when the ambient temperature ranged between 42 and 48 degrees C (hot-dry). The influence of hot climate in accident causation was evident from the shift-wise variations in the occurrence of accidents. The longitudinal study showed that the night workers were more vulnerable and less tolerant to heat, the tolerance time being less by about 15% (31 degrees C ET) to 40% (39 degrees C ET), compared to the rotating day workers. The relationship of the segmental and compartmental temperatures (segmental triggering response) played a critical role in heat dissipation/accumulation mechanism, and reflected in the heat tolerability of the day and night workers. PMID- 14564877 TI - Shift work at a modern offshore drilling rig. AB - The oil and gas exploration and production offshore units are classified as hazardous installations. Work in these facilities is complex, confined and associated with a wide range of risks. The continuous operation is secured by various shift work patterns. The objective of this study was to evaluate how offshore drilling workers perceived shift work at high seas and its impacts on their life and working conditions. The main features of the studied offshore shift work schedules are: long time on board (14 to 28 days), extended shifts (12 hours or more per day), slow rotation (7 to 14 days in the same shift), long sequence of days on the night shift (7 to 14 days in a row) and the extra-long extended journey (18 hours) on shift change and landing days. Interviews revealed a wide range of stressors caused by the offshore shift work, as well as difficulties to conciliate work with family life. It was observed that changes of the family model, leading to role conflicts and social isolation, work in a hazardous environment, perceiving poor sleep when working at night shifts and the imbalance between the expected and actual rewards are the major stressors for the offshore drilling workers. PMID- 14564878 TI - How a small enterprise improved the conditions of night and shift work using local resources. AB - A small oxygen factory in Cantho Province located in the Mekong Delta Area in the southern part of Vietnam was studied to provide practical support measures to improve night and shift work. A direct observation study and a fatigue symptom survey during the work were conducted. The factory applied discontinuous two shift systems in two teams. Depending on customers' demands, they frequently prolonged oxygen production until midnight. The study results showed work-related risks such as carrying heavy oxygen cylinders, workers' sleepiness during the night work, and increased fatigue feelings among production operators. Based on the study results, better strategies for night and shift work schedules such as regular work hours minimizing overtime and night work were discussed with the managers and workers. A follow-up visit three months later confirmed many improvements undertaken in the factory. Better work arrangements for night and shift workers were made including local lighting, resting corners, filling the height gaps on the work floors, and clear work instructions. Prolonged mid-night shift was stopped. It was concluded that local small enterprises in Vietnam have much potential to improve their conditions of shift and night work once practical support measures based on their local practice is given. PMID- 14564879 TI - Integrated ergonomics approach toward designing night and shift work in developing countries based on experiences in Bali, Indonesia. AB - Recently, the effort in carrying out an integrated ergonomics approach known as "SHIP" (systemic, holistic, interdisciplinary and participatory) approach has been intensively undertaken in Bali with the aim of sustaining improvements being done. The People's Consultative Assembly of the Republic of Indonesia issued for the 1999-2004 period a "SHIP" Act on the Macro Guidelines of Tourism Development in which ergonomics and other factors must be considered comprehensively to attain sustainable development in tourism. Therefore the night and shift work that is recently increasingly applied in the tourism industry must also be designed and organized through this approach. In fact, however, economic factors have still been the predominant reason for workers to accept any type of night and shift work decided by the management, without taking into account possible impacts and consequences. For example, rapid forward rotation schemes seem more adapted to the hotel industry instead of traditional 6-6-6 rotation. Further, inter-city bus drivers are approved to work a 24-hour shift followed by one day off. These drivers often work an additional risky night shift after two consecutive night shifts so as to meet needed expenses for the family. Cultural or religious activities still presented constraints for workers as they carried out subsequently the night work. Therefore, proactive steps should be taken in a timely manner through the integrated SHIP approach in designing night and shift work so as to achieve work schedules compatible with both social life of shiftworkers and business concerns. PMID- 14564880 TI - Support measures to improve night and shift work conditions in Thailand: a case study in a glass factory. AB - The present study aimed to examine the working conditions of shift workers in a multinational enterprise in Thailand and to identify practical support measures for improvements. A multinational, glass-manufacturing factory employing 1,500 workers was selected as the research site. Three shift systems in three teams were adopted. A direct observation study and a fatigue feeling monitoring study were carried out to compare the differences between different shifts. A 10-day time-budget study was conducted for 30 shift workers to know their work and sleep patterns. The direct observation study identified safety and health risks during the night work periods. The risks included insufficient lighting, height gaps on the floor, excessive exposure to heat, inappropriate workstations, and sleepiness and fatigue feelings among shift workers. Working consecutive double shifts and overtime work was often seen. An advisory meeting was held based on the study findings to assist managers and workers in improving their working conditions. A follow-up visit six months later confirmed that the glass factory implemented several improvements to help night and shift workers. It was concluded that the direct observation methods associated with the time-budget study were helpful for identifying practical action points and strengthening workplace initiatives. PMID- 14564881 TI - Measurement properties of the Shiftwork Survey and Standard Shiftwork Index. AB - The Shiftwork Survey (SS) was introduced, along with the Standard Shiftwork Index (SSI), to provide a set of standardized self-report measures to be used in shiftwork research. However, beyond the initial assessment, no attempt has been made to examine the measurement properties of these scales in an independent sample of shiftworkers. Our goal, therefore, was to examine the measurement properties of these scales in an industrial sample of primarily male shiftworkers (N = 370). We found that all scales had acceptable reliabilities (alphas). The confirmatory factor analyses revealed that the chronic fatigue, coping, job satisfaction, and sleep scales are the weakest psychometrically, and the anxiety, personality (extraversion, neuroticism), general health, and physical health scales are the strongest psychometrically. Using item response theory analyses, we found that the scales overall are generally adequate measures of their underlying constructs, although many items should be altered or omitted. Our results, however, are limited by reliance on a single sample. PMID- 14564882 TI - A 6-hour working day--effects on health and well-being. AB - The effect of the total amount of work hours and the benefits of a shortening is frequently debated, but very little data is available. The present study compared a group (N = 41) that obtained a 9 h reduction of the working week (to a 6 h day) with a comparison group (N = 22) that retained normal work hours. Both groups were constituted of mainly female health care and day care nursery personnel. The experimental group retained full pay and extra personnel were employed to compensate for loss of hours. Questionnaire data were obtained before and 1 year after the change. The data were analyzed using a two-factor ANOVA with the interaction term year*group as the main focus. The results showed a significant interaction of year*group for social factors, sleep quality, mental fatigue, and heart/respiratory complaints, and attitude to work hours. In all cases the experimental group improved whereas the control group did not change. It was concluded that shortened work hours have clear social effects and moderate effects on well-being. PMID- 14564883 TI - Job stress, social support at work, and insomnia in Japanese shift workers. AB - A cross-sectional study was conducted to clarify the contribution of psychological job stress to insomnia in shift workers. A self-administered questionnaire concerning job stress, sleep, depressive symptoms and lifestyle factors was submitted to a sample of 530 rotating shift workers of age 18-59 years (mean age 27) in an electric equipment manufacturing company. Perceived job stress, i.e., job demands, job control and social support at work, was assessed using the Japanese version of the Job Content Questionnaire. Insomnia was regarded as prevalent if the workers had at least one of the following symptoms in the last year; less than 30 minutes to fall asleep, difficulty in maintaining sleep, or early morning awakening almost everyday. Overall prevalence was 37.8%. Logistic regression analyses while adjusting relevant factors showed that lower social support at work was significantly associated with a greater risk of insomnia than the higher social support (adjusted OR 2.5). Higher job strain with lower social support at work increased the risk, compared to lower strain with higher support at work (crude OR 1.8; adjusted OR 1.5). Our findings suggest the low social support at work independently associated with insomnia in shift workers. PMID- 14564884 TI - Sleepiness in a population of Italian shiftwork policemen. AB - The aim of this study was to evaluate the effects of shiftwork on sleepiness, sleep disorders and sleep related accidents in a population of policemen. Data concerning age and physical characteristics, working conditions, sleep problems and accidents were collected by a questionnaire. Sleepiness was evaluated by the Epworth Sleepiness Scale (ESS) while the presence of sleep disorders was evaluated by a score (SD-score) drawn from indicators of insomnia, breathing disorders, periodic limb movements-restless leg syndrome and hypersomnia. The effects of age, gender, body mass index, working condition and seniority on ESS, SD-score and accidents were analysed by linear and logistic regression. Participants were 1280 policemen: 611 shiftworkers and 669 non-shiftworkers. The ESS scores were not higher in shiftworkers than in non-shiftworkers, but the SD score was found to be significantly influenced by shiftwork condition and seniority. The occurrence of sleep-related accidents was found to have been significantly increased for shiftworkers and related to the presence of indicators of sleep disorders. The sleepiness could be underestimated or even overcome by the influence of stressing conditions. However our data should alert occupational health physicians for the diagnosis and prevention of possible lurking intrinsic sleep disorders likely to influence health problems and risk of accidents in shiftworkers. PMID- 14564885 TI - Shiftwork locus of control effects in police officers. AB - Personality and experientially-based differences are suggested to moderate the negative effects of shiftworking. This investigation was one of a series of studies aimed at examining the relationship between internal locus of control (internality) and commonly reported outcomes of exposure to shiftworking. The study concentrated on whether or not shiftwork-specific internality appeared to be associated with better experiences for two groups of shiftworkers on different shift rotas. The Shiftwork Locus of Control (SHLOC) scale was completed by each group along with a range of outcome measures (including: sleep disturbance, alertness on shift, psychological well-being, disturbance of social and family life, and fatigue). The analyses controlled for for age, shiftwork experience and years of service effects. The results showed that for the majority of outcome variables, the higher internality was associated with significantly lower levels of shiftwork problems, especially when compared to the low internality group. The findings of this study further support the use of shiftwork-specific locus of control in shiftwork research as an indicator of potentially better tolerance, and possibly as part of an occupational health monitoring process to aid the targeting of interventions. PMID- 14564886 TI - Daytime cardiac autonomic activity during one week of continuous night shift. AB - Shift workers encounter an increased risk of cardiovascular disease compared to their day working counterparts. To explore this phenomenon, the effects of one week of simulated night shift on cardiac sympathetic (SNS) and parasympathetic (PNS) activity were assessed. Ten (5m; 5f) healthy subjects aged 18-29 years attended an adaptation and baseline night before commencing one week of night shift (2300-0700 h). Sleep was recorded using a standard polysomnogram and circadian phase was tracked using salivary melatonin data. During sleep, heart rate (HR), cardiac PNS activity (RMSSD) and cardiac SNS activity (pre-ejection period) were recorded. Night shift did not influence seep quality, but reduced sleep duration by a mean of 52 +/- 29 min. One week of night shift evoked a small chronic sleep debt of 5 h 14 +/- 56 min and a cumulative circadian phase delay of 5 h +/- 14 min. Night shift had no significant effect on mean HR, but mean cardiac SNS activity during sleep was consistently higher and mean cardiac PNS activity during sleep declined gradually across the week. These results suggest that shiftwork has direct and unfavourable effects on cardiac autonomic activity and that this might be one mechanism via which shiftwork increases the risk of cardiovascular disease. It is postulated that sleep loss could be one mediator of the association between shiftwork and cardiovascular health. PMID- 14564887 TI - Blood pressure and heart rate variability in workers of 8-hour shifts. AB - This study examined the effects of shiftwork on the cardiovascular system. The blood pressure (BP) and heart rate variability (HRV) of 134 male workers, who worked 8-hour shifts with rapid rotation of shifts at 3-day intervals, were examined for all the three shifts. In addition, the job stress was measured by Karasek's JCQ 49-item questionnaire and the circadian type was assessed by the morningness-eveningness questionnaire. The smoking and alcohol drinking habits, marital status and past medical history were also obtained. The method of analyzing the measured data based on a mixed model was used to illustrate the association between the shiftwork duration and the BP or HRV. The average age of workers was 29 years (between 25-44). Among them, 77.9% were current smokers, 50% showed the passive type of job strain in Karasek's model. The mean shiftwork duration was 5.21 years (range 5.4 months--10 years). In the circadian type, none of them belonged to a definitely morning type or a definitely evening type. In the multivariate analysis adjusted by age, job strain, shift, circadian rhythm and smoking, the blood pressure showed significantly increasing trends according to shiftwork duration in both the systolic and diastolic BP. The heart rate variability also showed a significantly decreasing trend according to the shiftwork duration in both the parasympathetic and sympathetic functions (p < 0.05). These results suggests that there are negative health effects arising from shiftwork on the cardiovascular system. PMID- 14564888 TI - Heart rate variability during long truck driving work. AB - We recorded ambulatory electrocardiograms of 6 long distance truck drivers during their work period in order to observe the affect of autonomic nervous function and symptoms while doing their work. We also recorded their work patterns every minute. The RR50 value and the LFP/HFP ratio were calculated every two minutes based on R-R interval data. RR50 was significantly higher during taking naps than during other periods of work shifts, while, the LFP/HFP ratio showed significantly lower during taking naps than during other periods of work shifts. RR50 in the morning was significantly higher than that in the afternoon. On the contrary, the LFP/HFP ratio showed opposite tendency. Only on the times of driving, RR50 was significantly higher in the morning than that in the afternoon. On the other hand, the LFP/HFP ratio showed an opposite tendency. These results show that the parasympathetic nervous activities were more dominant than sympathetic nervous activities in the morning during the subjects were doing long distance truck driving including midnight work. Driving while in high parasympathetic nervous activity levels may add to cardiovascular stress and lead to drowsiness. And this may result in disrupted attention. It is necessary to decrease work time and improve working conditions of truck drivers working long hour shifts. PMID- 14564889 TI - Ultradian rhythms in processing speed of laterally exposed words and pictures. AB - The study was designed to find out the cerebral hemispheres oscillations in stimuli processing during the 24-hour period of wakefulness in isolated subjects remaining in a monotonous environment. Stimuli processing speed from the 24-hour constant routine periods (06.00-06.00 h) of a larger experiment were analysed for the purpose of this paper. Parallel sets of words and pictures were exposed laterally using a purpose-designed computer program. The subjects reacted to pictures or words by pressing appropriate buttons. The significant dominant ultradian rhythms (around 4 h) in the processing speed of words addressed to the right hemisphere were found and of pictures addressed to the left hemisphere. Longer significant dominant periodicities (around 12 h) appeared in the processing speed of words addressed to the left hemisphere and of pictures (around 8 h) addressed to the right hemisphere. Ultradian rhythmicity of the central nervous system functioning is suggested. PMID- 14564890 TI - Circadian temperature rhythms in clockwise and counter-clockwise rapidly rotating shift schedules. AB - The purpose of this study was to examine the circadian temperature rhythm in clockwise (CW) and counter-clockwise (CCW) rapidly rotating shift schedules. Arguments against the CCW rotation of shifts are that they result in shortened sleep and promote greater disruption of circadian rhythms. The 3-week study included a week of day shifts (0800-1600) and 2 weeks of shiftwork. The CW 2-2-1 schedule rotated from two early mornings (0600-1400) to two evenings (1400-2200) to one midnight shift (2200-0600) allowing 24 hours off at each shift rotation and a 48-hour weekend. The CCW schedule rotated from two evenings to two early mornings to one midnight shifts allowing only 8 hours off at each shift rotation and an 80-hour weekend. Analysis of the 72-hr periods at the end of each workweek, including the midnight shifts and recovery periods during weeks 2 and 3 were compared to the same 72-hour period at the end of week 1 (baseline). A cosine function that fit the temperature curves by minimizing the sums of squares produced parameters that underwent analysis of covariance procedures. Significant differences were found between rotation conditions for amplitude and acrophase. An attenuation of amplitude and a delay in the acrophase was the found for the counter-clockwise condition. Features inherent in this schedule might explain these effects, particularly, the increased opportunity for "sleeping in" at the beginning of the week and an expanded (2-shift) workday at the end of the week. PMID- 14564891 TI - Physiological effects of shift work on hospital nurses. AB - OBJECTIVE: The purpose of this study was to assess the physiological effects of shift work on the urinary excretion rates of norepinephrine, 6-sulfatoxymelatonin and estriol in hospital nurses. METHOD: Twenty-four hour urine specimens were examined on a daytime/nighttime basis for each work shift of pregnant and non pregnant subjects. The urinary norepinephrine and 6-sulfatoxymelatonin were measured by enzyme-linked immunosorbent assay and estriol by radio-immunoassay. RESULTS: Urinary norepinephrine level during the night work was higher than the night levels of the days off and the day shift. Urinary 6-sulfatoxymelatonin level during the night work was lower than the night levels of the days off and the day shift. Urinary estriol level of pregnant subjects showed no differences among work shift and also between daytime and nighttime. CONCLUSIONS: Urinary excretion rates of norepinephrine and 6-sulfatoxymelatonin were affected by shift work both for non-pregnant and pregnant subjects. It was unlikely that urinary estriol levels in the pregnant subjects were significantly affected by shift work. PMID- 14564892 TI - A week of simulated night work delays salivary melatonin onset. AB - In most studies, the magnitude and rate of adaptation to various night work schedules is assessed using core body temperature as the marker of circadian phase. The aim of the current study was to assess adaptation to a simulated night work schedule using salivary dim light melatonin onset (DLMO) as an alternative circadian phase marker. It was hypothesised that the night work schedule would result in a phase delay, manifest in relatively later DLMO, but that this delay would be somewhat inhibited by exposure to natural light. Participants worked seven consecutive simulated 8-hour night shifts (23:00-07:00 h). By night 7, there was a mean cumulative phase delay of 5.5 hours, equivalent to an average delay of 0.8 hours per day. This indicates that partial circadian adaptation occurred in response to the simulated night work schedule. The radioimmunoassay used in the current study provides a sensitive assessment of melatonin concentration in saliva that can be used to determine DLMO, and thus provides an alternative phase marker to core body temperature, at least in laboratory studies. PMID- 14564893 TI - The effect of a change in sleep-wakefulness timing, bright light and physical exercise interventions on 24-hour patterns of performance, mood and body temperature. AB - Experiments consisting of baseline, bright light and physical exercise studies were carried out to compare the effect of a 9-hour delay in sleep-wakefulness timing, and the effects of bright light and physical exercise interventions on 24 hour patterns of performance, mood and body temperature were examined. Each study comprised a 24-hour constant routine at the beginning followed by 3 night shifts and 24-hour constant routine at the end. Performance on tasks differing in cognitive load, mood and body temperature was measured during each constant routine and the interventions were applied during the night shifts. The 24-hour pattern of alertness and performance on the tasks with low cognitive load in post treatment conditions followed the change in sleep-wakefulness timing while more cognitively loaded tasks tended to show a reverse trend when compared to pre treatment conditions. There was a phase delay around 4 hours in circadian rhythms of body temperature in post-treatment conditions. PMID- 14564894 TI - Psychophysiological effects of a brief nocturnal light exposure. AB - This study compared the effects of a brief pulse (60-minute) of three full spectrum light intensities (1000, 500 and 30 lux) and two green light intensities (1000 and 500 lux) administered between 0200 and 0300 hrs. Ten participants were involved in this repeated measures study. Each participant experienced one condition every week for five weekends. Sessions began at 1800 hours and ended at 0600 hours the following day. Outside of the 60-minute exposure period, each session was spent in 30 lux white light. Oral temperature, salivary melatonin, cognitive performance and subjective mood were sampled throughout the sessions. Analysis revealed that all of the experimental light conditions significantly reduced salivary melatonin concentrations immediately following the pulse. This effect was not maintained beyond the duration of the light pulse. There was no significant effect on oral temperature. There were also no significant effects on cognitive performance and subjective mood, though some positive trends were observed. These results argue that brief, moderate intensity, pulses of either green or full spectrum light are sufficient to suppress the normal nocturnal rise in melatonin. However, the level of suppression obtained does not translate into significant improvement in cognitive performance or subjective mood. PMID- 14564896 TI - Evaluation of a fatigue model using data from published napping studies. AB - The authors have previously published the development and empirical validation of a work-related fatigue model. However, published work has not involved data from napping studies. The aim of this paper is to determine how closely the model predicts changes in subjective and objective measures from data published from napping studies. The regression results between the model outputs and logical reasoning, multiple sleep latency test scores, self-rated alertness, profile of mood state fatigue, visual vigilance and reaction time were all strong to very strong (R2 = 0.4-0.9). Only digit symbol substitution revealed moderate (R2 = 0.1 0.2) regression values. The outputs of the model reflect changes due to naps of varying duration and timing measured at varying periods following a nap. Together with the outputs from previous investigations, these results further support the potential use of the fatigue model in operational settings. This appears to be true in settings that utilise napping as well as those that do not. PMID- 14564895 TI - The effects of bright light and nighttime melatonin administration on cardiac activity. AB - Although melatonin has an important physiological role in the facilitation of sleep, its precise mechanism of action is not clear. To investigate the potential contribution of melatonin to influence cardiac autonomic activity in the evening, 16 young healthy subjects participated in a repeated measures design where cardiac autonomic activity, heart rate and blood pressure were examined during three experimental conditions. An initial baseline condition involved dim light exposure (< 10 lux), permitting the normal nocturnal rise in endogenous melatonin. In other sessions, subjects were exposed to bright light (> 3000 lux) to suppress melatonin secretion and administered a placebo or melatonin (5 mg) capsule at the estimated time of increase in endogenous melatonin (wake time + 14 hours). Heart rate, pre-ejection period (a measure of cardiac sympathetic activity) and respiratory sinus arrhythmia (a measure of parasympathetic activity) were not significantly altered in response to the three melatonin levels. While melatonin had no effect on diastolic blood pressure, systolic blood pressure was maximally decreased by 6 +/- 1.93 mmHg (mean +/- SEM, p < 0.005) 150 minutes after exogenous melatonin. The results indicate that melatonin does not directly modulate cardiac autonomic activity, but may rather act directly on the cardiovascular system. PMID- 14564897 TI - Shiftwork, age and well-being: recent developments and future perspectives. AB - The working population is aging fast in most European countries and in the USA. The health and well-being of an elderly shift worker depends on the interaction of several individual, medical, psychosocial and job-related factors. These factors are related to the biological ageing process, but also to changes in one's individual life situation and the needs of the ageing shift worker. The evidence of good age-specific solutions in working hours is limited, but the few published intervention studies support the use of individual flexibility, rapid forward rotating shift systems, and earlier shift start-end times in three-shift work. In addition to the development of shift schedules, the counter-measures to improve the health and well-being of ageing shift workers should be focused on the improvement of occupational health care and the promotion of appropriate coping mechanisms for the aging. PMID- 14564898 TI - Different job demands of nightshifts in hospitals. AB - It is now recognized that the effects of nightwork result jointly from desynchronization of circadian rhythms and from the job content. In scheduling nightwork, use should be made of the shift systems most compatible with the biological rhythms and social patterns of the workers concerned, and specific measures should be implemented to reduce demands on nightworkers. Job content is of particular importance in the hospital sector, where demands vary greatly from one department to another. Above all, the tasks accomplished by those on nightshift differ markedly from those of daytime workers: nightwork is limited to scheduled health care jobs that cannot be deferred until the morning, and to management of incidents. Yet staffing levels at night are reduced. Certain daytime resources are lacking at night, and the working conditions, generally defined with reference to the activities of daytime personnel fail to take into account the specific features of nightwork which are largely ignored by the management. On the basis of various studies conducted in French hospitals (extensive questionnaire surveys, ergonomic analyses in different departments), these features of nightwork and the corresponding need for appropriate responses, in terms of shift systems and job content, are reviewed and discussed. PMID- 14564900 TI - Age and the subjective experience of shiftwork. AB - The objective of this study was to examine age related effects of shiftwork albeit difficult to tease apart the natural effects of aging, and lifestyle or behaviour, or job done, over time and the shifts a person works. This is an issue of concern because the numbers of shiftworkers over 45 are increasing. Participants were 306 police officers who had worked a new rota for approximately 6 months. Three age groups were compared (1 = 20-32.9, 2 = 33-39.9, 3 = 40+) using a range of shiftwork-related measures and multivariate analysis of covariance (controlling for shiftwork experience and other individual differences). Younger officers tended to report significantly better attitudes towards their shiftwork, better adjustment to night-bound shifts, greater job satisfaction and organisational commitment, lower fatigue and longer sleep durations. Older shiftworkers reported significantly higher morningness and lower sleep need than the younger officers. This concurred with existing research that implicates such variables in the mechanism(s) involved in age-related tolerance to shiftwork. It was also evident that the older group tended to resort to greater caffeine intake on all shifts. The findings offer tentative support for the position that age can be linked to depleted shiftwork tolerance but the issue of establishing the relative impacts of aging, lifestyle, behaviour, work type and the rota worked remains to be a challenge. PMID- 14564899 TI - Beliefs about age and physical demands of work in shiftworkers. AB - The aim of the study was to investigate the differences between shiftworkers and non-shiftworkers in terms of several aspects of aging. Our interest was in particular focused on the differences observed in the assessment of subjective age, and physical, mental and social demands of work. Subjects were workers in a medical setting, 54 shiftworkers and 53 non-shiftworkers of the comparable chronological age. Subjective age measures included cognitive age, desired age and beliefs about age. The results showed that shiftworkers, compared to non shiftworkers, reported being more physically tired by their work and that their work required more physical effort. No differences were found in reports of mental and social effort or tension produced by work. According to our data on subjective age measures, the subjects in either group felt approximately five years younger than their chronological age, desired to be ten years younger and believed that the person who is two years younger than them is most successful in its job. There were no differences between shift- and non-shiftworkers in terms of cognitive age, but shiftworkers were inclined to choose younger age as their desired age, and believed that younger people are more successful in their work. PMID- 14564901 TI - Evaluation of a fast forward rotating shift schedule in the steel industry with a special focus on ageing and sleep. AB - The aim of this study was to explore the effect of a change in the speed and direction of shift rotation on the sleep and wakefulness of younger and older workers. A continuous three-shift schedule was changed from a slow backward rotating (EEE-MMMNNN-----) to a fast forward rotating system (MMEENN----). Sixteen subjects (mean age 42 years) were studied before and one year after the change in schedule. Two age groups were compared: ten younger men (mean age 35 years) and six older men (mean age 53 years). The effects of the new work schedule were evaluated by a questionnaire (modified SSI), and on-site registrations with an actigraph and sleep log for one shift cycle (10-15 days) before and after the new schedule. After the change in schedule, subjective sleep problems decreased and alertness increased during the morning shifts. The change in schedule influenced sleep differently in the two age-groups. Both the subjective and objective quality of sleep improved among the older workers. The results indicate that a fast forward rotating shift schedule is more suitable for older workers than a slower backward rotating system. PMID- 14564902 TI - Searching for preventive measures of cardiovascular events in aged Japanese taxi drivers--the daily rhythm of cardiovascular risk factors during a night duty day. AB - Previous studies have shown that Japanese taxi drivers are exposed to more risk factors and have a higher mortality rate due to cardiovascular disease than other occupational groups. We investigated the effect of night taxi driving with a view to preventing acute events of cardiovascular disease among aged taxi drivers. Twenty-nine taxi drivers (41-67 years old) were examined for urine normetanephrine/creatinine, von Willebrand factor, anti-thrombin III, t plasminogen activator-plasminogen activator inhibitor 1-complex, hematocrit, blood glucose and blood pressure in the morning and at midnight during a duty day and in the following morning. At the same time, the blood pressure and blood glucose of 46 taxi drivers (43-67 years old) in the morning after a night duty with little sleep and in the morning after daytime work and subsequent night sleep were compared. The results obtained indicate that the aggravation of sympathetic nervous system functions with disturbed circadian rhythms, increased blood coagulation and blood concentration, endothelial injury and the elevation of blood glucose at midnight or the next morning were induced by their night work. These conditions are supposed to favour acute vascular events in aged taxi drivers. Preventive measures considered include social support for anticoagulant food and water intake, short exercise and walking as well as taking a rest and a nap during night work. PMID- 14564903 TI - Work schedules in health care in France: very few changes between 1991 and 1998, according to national data. AB - It is important to know whether working time schemes offer, following the adoption of the 1990 ILO Night Work Convention, a better use of the actual knowledge in moderating the adverse effects of shiftwork for both sexes. The last two national studies on a representative sample, about working conditions in France (1991, 1998), show an increase of night work among female nursing staff. Long hours of work are becoming more common for nurses. The percentage of women health care workers not working the same number of days every week increased, with work weeks of 7 days or more. Few positive aspects exist such as a reduction in the proportion of those having to work 17 Sundays or more and a reduction of shifts beginning before 7 a.m. for female nurses. New negotiations, starting in 2001, should be an opportunity for improvements. PMID- 14564904 TI - Gender and sleep in nightworkers: a quantitative analysis of sleep in days off. AB - Differences in sleep patterns between workdays and days off contribute to shiftwork effects on workers' health and well-being. But regardless of shift schedules, female workers face more difficulties in fulfilling their sleep need because of housework. This study analyzes gender differences concerning sleep in days off by comparing sleep patterns in male and female nightworkers, analyzing sleep as related to the presence of children and testing the association of sleep features between workdays and days off. Male (n = 16) and female (n = 30) workers at a plastic plant, working from 10 p.m. to 6 a.m., on weekdays, filled sleep logs for seven consecutive weeks. Male and female samples did not differ in length of night sleep or in total length of sleep. For both samples, sleep length/day in days off increased, but the difference was lager among females. Also important were the relations between sleep in workdays and days off, specially among women. Among female workers, the results indicated that workers with children tended to sleep less in Saturday mornings, suggesting a negative effect of motherhood on sleep not restricted to workdays. The general results indicate that sleep need on the one hand, and social factors on the other determine the actual amount of sleep. PMID- 14564905 TI - Effects of shift changes on female workers at a dish factory. AB - This study investigated the effects of working night shifts on social and family life by examining changes in workers' daily life before and after a change in their shifts. Subjects were 40 women aged 27-59 years, working at a dish factory. During the health examination of night workers in autumn of 2000, the subjects were directly interviewed about changes in their lives induced by the shift change. Question parameters consisted of 8 items including 30 sub-items related to social and family life, such as sleep, rest, meals, sports, family time, hobbies, neighborhood association and social activities. The subjects selected one of four response categories: "becoming worse", "no change", "becoming better" and "difficult to determine." With regard to the percentage of "becoming worse", meal-related items ranked high in all of the shift types. "Family time" and "hobbies" showed high percentages in the subjects transferring from day shifts to night shifts, and in those transferring from early-morning shifts to night shifts. "Rest", "sports" and "hobbies" showed high percentages in the subjects transferring from night shifts to midnight shifts. Decreased sleeping hours were confirmed in all of the shift types, while the subjects tended to sleep more soundly. As the workers transfer to shifts at earlier hours, they were obliged to make sacrifices in various aspects of their social and family life. Therefore, much assistance in this regard should be given to them. PMID- 14564907 TI - Effects of stress on psychophysiological parameters of electricity distribution network controllers in Ukraine. AB - The purpose was to reveal the effects of stress on the parameters of cognitive performance and cardiovascular system activities in controllers working 12-hour shifts. Sixteen controllers were studied, and altogether 384 subject observations were acquired. A 5-point scale was used to estimate the perceived level of stress experienced by the controllers. Increased heart rate and heart activity-related haemodynamic parameters were found with an increase in perceived stress. Decreases found in heart rate, circulatory minute volume, and Kerdo's vegetative index over the shift became less pronounced with increased stress. Better attention was found under the average level of stress that was found on the first day shift. An effect of stress on short-term memory was not revealed. As a whole, the most pronounced changes were found during the first day shift and less pronounced changes appeared during the first night shift. Effects of stress were not found in changes of the studied parameters during the second consecutive day or night shift. Thus, increased stress causes the activation of some psychophysiological functions (attention and cardiac activity) that are indispensable for high work efficiency under increased production demands. However, the necessary activation could not be maintained during the second consecutive 12-hour shifts, probably because of accumulated fatigue, and also night work. On the other hand, too high stress may lead to the excessive activation of cardiac activity and deterioration of attention. PMID- 14564906 TI - The impact of unwaged domestic work on the duration and timing of sleep of female nurses working full-time on rotating 3-shift rosters. AB - The study examined the impact of family type on the timing and duration of sleep of 16 experienced female shiftworkers working a rotating 3-shift roster. The nurses lived in one of three domestic lifestyle arrangements: single with no child care responsibilities (N = 4), partnered with no child care responsibilities (N = 5) and partnered with child care responsibilities (N = 7). Self report sleep diaries were used to collect data over a period of 28 days, following which each nurse took part in a conversational interview. Comparisons of the roster mean sleep durations between groups show that nurses who do not have the added unwaged workload of child care, record significantly more sleep than nurses with such responsibilities. Analysis of the data by shift type shows a significant difference for afternoon shift: nurses with child care responsibilities record a significantly earlier rise time and a significantly shorter total sleep duration. The interview data further highlights how sleep patterns are related to the time constraints of both domestic and waged work. PMID- 14564908 TI - Daily and yearly burnout symptoms in Israeli shift work residents. AB - Burnout is a syndrome of physical and emotional exhaustion that develops among individuals who are open to public demands. In view of their heavy work load and sleep deprivation, we decided to evaluate the impact of long working hours on burnout and psychological status among a sample of residents during the first 2 years of their residency. Seventy-eight residents participated in the study, all residents completed self-administered questionnaires, and their sleep-wake cycle was monitored by a wrist-worn actigraph for a period of 5-7 days. The questionnaires included a short form suitable for Experience Sampling Method (ESM), and a longer background Questionnaire. The results revealed that sleep duration, Work Load and the interaction between them, explain the Negative Mood the day after the night shift. However, positive mood, and fatigue were not affected by sleep duration or workload. In general, after one year of residency, residents become more stressed, less involved in the job, and had a high level of burnout and psychosomatic symptoms. However, after the second year, the burnout symptoms were almost the same as at the beginning except for the level of stress that remained high. Sleep duration was unrelated to the burnout symptoms. PMID- 14564909 TI - Effects of supervisor support and coping on shiftwork tolerance. AB - This study examines the effects of supervisor support and coping on work/non-work conflict and health in shiftwork. It describes a model of shiftwork tolerance that is tested on samples from two populations of shiftworkers. The samples--of nurses and ambulance workers--differed by occupation, gender and shift schedule. Quantitative (survey questionnaire) and qualitative methods (in-depth interviews) were used to triangulate results and to yield richer data on psychosocial variables. Structural equation modelling, using EQS, was used to describe the common path relationships observed within both samples of shiftworkers. The results demonstrated important relationships between social support from supervisors, coping strategies, work/non-work conflict and symptoms that transferred robustly between the two populations of shiftworkers. PMID- 14564910 TI - Work activities of practical nurses and risk factors for the development of musculoskeletal disorders. AB - Musculoskeletal and emotional disorders are important causes of reported diseases, causing medical absences, and eventually earlier decrease of work ability. This paper reports the results of a study carried out among practical nurses working at the Orthopedics and Trauma Institute. The objectives of the study were: (a) to describe the routine activities performed during day and night shifts, and (b) to compare the work activities performed in different wards during these shifts. A Brazilian version of the Work Ability Index--WAI (TUOMI et al., 1994) was answered by 83 practical nurses. Forty-three of them (52%) reported pains or musculoskeletal diseases, either based on their own opinion or diagnosed by a physician. These nurses were invited to join the second phase of the study and twenty-nine accepted it. All work activities performed in 29 shifts were observed and recorded. The results showed that day shifts were far more demanding in terms of the number of activities related to patients' care than afternoon and night shifts. Also, body postures associated with day work activities demanded important physical efforts. The number of nurses in charge during night shifts was substantially lower than during day shifts. This could lead to an overload and affect the health of the nurses. PMID- 14564911 TI - Can we consider medical residents as shift workers? AB - Although medical residents are characterized by long working hours, night shifts and high levels of work load, it is unclear if their work schedule can be classified as shift work, or if it has a similar impact on residents' well-being. The present paper compared the profile of complaints about sleep or daytime functioning of medical residents to that of rotating shift workers and day workers, of similar ages. Sixty-one residents (aged: 32.2 +/- 2.2 years), after 2 years of residency, participated in the study. The two control groups with a similar age range (26-40 years) were chosen, and included 94 rotating shift workers and 146 day workers. All subjects completed self-administered questionnaires on their sleep habits, and their sleep-wake cycle was monitored by a wrist-worn actigraph. Ten percent of the residents complained about difficulties falling asleep, 34% complained about morning tiredness, 14% complained about mid-sleep awakening, and 20% about prolonged fatigue. The residents slept significantly less than the day workers, and their sleep efficiency was significantly higher. When examining their subjective complaints profile, residents complained more than day workers and their answers were more similar to those of rotating shift workers, therefore they can be considered to be characterized as shift workers. PMID- 14564912 TI - Time of day type of food--relation to mood and hunger during 24 hours of constant conditions. AB - A six-day high-carbohydrate meal (HC; 65 E% (energy percent) carbohydrates, 20 E% fat and 15 E% protein) and a six-day high-fat meal (HF; 40 E% carbohydrates, 45 E% fat and 15 E% protein) were given to seven healthy subjects in a crossover design. On the last day subjects were kept awake for 24 hours in a metabolic laboratory while substrate utilisation and energy expenditure were measured by indirect calorimetry. The subjects were given isocaloric meals every four hours. Results showed that hunger decreased at night (F = 4.2, p < 0.05) and linearly increased after meal intake. Macronutrient composition (fat/carbohydrates) seemed to be of less importance for hunger. Hunger and thirst were found to be strongly associated with gastrointestinal substances, for hunger the strongest being a negative correlation with triacylglycerol (partial correlation = -0.39). It is suggested that it might not be necessary for shift workers to eat full portions at night but that satiation will occur with less food. Possibly lack of adjustment of nocturnal food intake might be one reason why overweight is common in shift work populations. PMID- 14564913 TI - A cross-sectional study of the health effects of work schedules on 3212 hospital workers in France: implications for the new French work schedules policy. AB - This study was designed to investigate the effects of work schedules on the health of hospital workers at the Assistance Publique-Hopitaux de Paris (AP-HP). Out of 40 hospitals, 17 volunteered to participate in this study. The Standard Shiftwork Index and a questionnaire concerning physicians' work schedules were used. Ten thousand questionnaires were distributed anonymously to hospital workers between March and April 1999. Professional categories comprised head nurses, nurses, nursing auxiliaries, hospital agents, midwives and full time physicians. Departments included internal and geriatric medicine, general paediatrics, orthopaedic and general surgery, operating and emergency rooms, and anaesthesiology and intensive care units. 3250 questionnaires were returned. Demographics for the respondents were: 79.2% female, average age 38.1 +/- 9.1 years old. Eleven work schedules were identified. One fourth of the personnel had fixed morning work schedules. The highest level of job satisfaction was found in personnel working in paediatrics while dissatisfaction was strongest in the gerontology and, emergency room personnel. General Health Questionnaire (GHQ) scores were high for head nurses, operating room nurses and junior doctors as well as for personnel with rotating and flexible shifts. This study will be used to make recommendations concerning the reduction of working time for French hospital workers. PMID- 14564914 TI - Sickness absence and shift work among Japanese factory workers. AB - To investigate the effect of shift work on long-term sickness absence (more than 7 calendar days), an 8-year follow-up study was carried out in a factory in Japan. The participants were male employees aged 18-54 years who were engaged in manufacturing sites. Shift patterns were classified by the number of non-daytime working days during the previous one-year. The causes of sickness absence were classified into three groups, 1) causes except injury, diseases of the musculoskeletal system and connective tissue, 2) injury, 3) diseases of the musculoskeletal system and connective tissue. The analysis of long-term sickness absences was based on the first occurrence. The age-adjusted incidence of sickness absence among shift workers who were on non-daytime shifts more than two thirds of working days during the previous one year was significantly higher than that among other workers. After adjusting for confounding factors, a significant high risk still existed for this group of workers taking sickness absence for all causes and causes except for injury and the musculoskletal disorders. These findings suggest that shift workers who are engaged on a particular shift schedule are more likely to take leave due to sickness. PMID- 14564915 TI - Comparisons of psychosomatic health and unhealthy behaviors between cleanroom workers in a 12-hour shift and those in an 8-hour shift. AB - The General Health Questionnaire (GHQ12) and physical fitness tests were administered to 338 workers in clean rooms producing electronic parts in 12-h shifts. The results were compared to those in 95 workers in 8-h shifts and 284 daytime management, clerical and engineering workers. The 12-h shift workers complained of poor health, dissatisfaction with life and poor recuperation from fatigue more than the 8-h shift workers although the rates of complaints were highest in the daytime workers. The GHQ scores were similar in the two shift groups, and much better than those in the daytime workers. However, the 12-h shift workers showed significantly lower fitness levels than the 8-h shift workers, and the levels were even worse than the daytime workers who had higher mean age and BMI levels compared with the shift workers. The tendency to have sedentary freetime activities and larger alcohol and cigarette consumption were observed in the 12-h shift workers. The 12-h shift work may have contributed to the unhealthy behaviors resulting in lower physical fitness levels. Health promotion services at the workplace should devote greater attention to long-hour shift workers, together with devising the ways to improve working conditions and environments for reducing fatigue at work. PMID- 14564916 TI - Health conditions of bus drivers in a 6 year follow up study. AB - The bus drivers of a public bus company, working in a fast rotating 4 shift system from 05.00 to 24.30, were examined in 1993 (230 persons) and 1999 (266 persons). The comparison between the two years showed no significant differences for all the parameters evaluated by the Standard Shiftwork Index and medical examination. In both surveys work organisation was considered "efficient-fairly good" by most workers, who were mostly satisfied with their job. Work load was rated significantly higher for "afternoon" and "morning" shifts, during which most accidents at work and "in itinere" occurred. Night sleep was reduced by 3 hours on "early" shift and about 2 hours on "morning" shifts. The most prevalent health troubles dealt with low back pain, gastritis, headache and haemorrhoids. Neuroticism was the trait more correlated with poorer health conditions, whereas shiftwork exposure appeared as a significant predictor of risk of critical Effort/Reward Imbalance and minor psychological disorders. Both extrinsic and intrinsic efforts significantly increased with age, but not reward. The comparison of the same cohort of 108 persons examined both in 1993 and in 1999 showed a significant increase of low back pain, gastrointestinal troubles, haemorrhoids and lipids disorders. PMID- 14564917 TI - Traumatic diaphragmatic hernia: delayed presentation seven years after penetrating trauma. PMID- 14564918 TI - The social impact of medicine. Chapter 11. PMID- 14564919 TI - Commentary on physician regulation in Delaware. PMID- 14564920 TI - Delaware Academy of Medicine: honoring traditions, securing the future. PMID- 14564921 TI - [Noroviruses are the most common pathogens causing nosocomial gastroenteritis outbreaks in Hungarian hospitals]. AB - INTRODUCTION: Norovirus (genera; previously "Norwalk-like viruses") of the family Caliciviridae are an important cause of acute gastroenteritis outbreaks. Epidemics commonly occur in crowded community and spread by fecal-oral route through contaminated food, water and direct contact. AIMS: To determine the incidence and epidemiological role of caliciviruses in nosocomial gastroenteritis outbreaks in hospitals in Hungary. METHODS: Between November 1998 and April 2002, 277 stool samples of 39 nonbacterial hospital outbreaks reported as "enteritis infectiosa" with unknown etiology from 13 of 19 counties in Hungary were examined by reverse transcription-polymerase chain reaction (RT-PCR) for Norovirus and Sapovirus and by a new recombinant enzyme immune assay (rEIA) for Norovirus only. RESULTS: 99 (36%) of 277 stool specimens were positive by RT-PCR and 29 (74%) and 38 (97%) of 39 outbreaks were positive by RT-PCR and RT-PCR and/or rEIA for Norovirus, respectively. Norovirus were the most frequently detected agents in the registered nosocomial enteric hospital outbreaks in years 2000 (n = 4; 31%) and 2001 (n = 7; 23%). In the first 4 months of 2002, 25 (66%) of 38 nosocomial outbreaks was caused by Norovirus. Seventeen (44%) outbreaks occurred in single internal wards and eight (21%) in more than one (2-8) ward. An average of 32% (0 80%) of the sick persons, were member of the hospital staff. Twenty four (63%) Norovirus-associated outbreaks occurred between January and March 2002. CONCLUSIONS: This is the first comprehensive country-wide surveillance of nosocomial gastroenteritis outbreaks in hospitals associated with Norovirus. Data showed that Norovirus were the most important cause of these outbreaks in Hungary but the epidemiological role of this virus is probably further underestimated. PMID- 14564922 TI - [Experiences in the invasive treatment of acute coronary syndrome]. AB - INTRODUCTION: Primer percutaneous coronary intervention is a very powerful tool in the treatment of acute coronary syndrome. The aim of the authors was to work out and analyse the methods of making the upto-date percutaneous coronary intervention available for patients living far from the Heart Institute. PATIENT AND INTERVENTIONS: Between 1st January 2000 and 31st October 2002, 221 patients with acute coronary syndrome were sent to intensive treatment from Kaposvar to the Heart Institute in Pecs partly by helicopter. The average age of patients was 54 years. 103 of them with acute myocardial infarction and 118 others with unstable angina were catheterised. Revascularization was achieved in 133 cases, and coronary operation in 63 cases. Primary intensive therapy was applied on 34 patients with infarction. RESULTS: No lethal complications arose during transport or operation. Mortality rate coming from cardial complications was only 4% during the first two years. These results are based on well organised cooperation between the Department of Internal Medicine Kaposvar and Heart Institute Pecs. CONCLUSION: Given suitable logistic background and adequate indication the risk of transporting patients can be taken. The percutaneous coronary intervention proved to be successful. PMID- 14564923 TI - [Studies on the novel association of human herpesvirus-7 with skin diseases]. AB - INTRODUCTION AND AIM: Human herpesvirus 7 in pityriasis rosea, this and other viruses in papular-purpuric gloves-and-socks syndrome have been implicated, but their primary or recurrent infections are still in question. PATIENT AND METHODS: In one available blood sample, therefore, IgM, IgG and its high avidity fraction characteristic for recurrent infections were quantitated by indirect immunofluorescence. Peripheral lymphocytes were subjected to nested polymerase chain reaction to detect viral DNA, or cocultivated with several cell cultures. RESULTS: One third of 33 pityriasis rosea patients had elevated IgM, another third had elevated IgG without high avidity molecules to human herpesvirus 7 suggesting primary infection. Thirty percent of controls, more than half of the patients had virtual DNA in their lymphocytes, but only one in 5 skin biopsy specimens were PCR positive. All three co-cultivation attempts yielded viruses extremely rapidly, verified by electron microscopy, polymerase chain reaction and monoclonal antibodies as human herpesvirus 7. These are the first isolates in the geographical regions of Hungary. These data suggest that pityriasis rosea is the consequence of a primary human herpesvirus 7 infection in seronegative adults, and only occasionally is due to virus reactivation. One patient with gloves-and socks syndrome had an acute, another patient had a persistent coinfection with human herpesvirus 7 and parvovirus B19, two others had a primary herpesvirus 7 infection. Interestingly, this disease might be elicited by both viruses individually or in synergism. CONCLUSION: Neither human herpesvirus 7 nor parvovirus B19 infect skin cells, but both can be detected in the infiltrating lymphocytes of skin eruptions, in which they induce an altered mediator production, that might be responsible for the general and local symptoms. PMID- 14564924 TI - [Cross-sectional survey on the nutritional and life-style habits of secondary school students in Hungary]. AB - INTRODUCTION: The national representative survey of secondary school students (age: 15-18 years) was undertaken between 1997 and 2000. The authors presented the data related to the nutritional and life-style habits. METHODS: The data for 6,347 students (48.4% boys and 51.6% girls) were evaluated. The anonymous questionnaire contained inquiries on nutritional rhythm, food frequency, use of vitamin and mineral supplements, smoking, alcohol and drug consumption habits, regular physical activity and family background (qualification, smoking, alcohol consumption). Within the frame of personal interviews the authors collected data on students' history with regard to occurrence of diabetes mellitus, other chronic diseases and in girls the time of the first menstruation. Concerning parents' history the authors collected information on the incidence of hypertension, myocardial infarction, diabetes mellitus, and stroke. STATISTICAL EVALUATION: Data were recorded in Data-Ease database system, in forms of individual records. The individual records were converted to be suitable for the statistical evaluation by SPSS for windows 9 statistical program pack. Data are presented as means and percent distribution. RESULTS: The nutritional rhythm of students was irregular. The daily consumption of milk, dairy products, fruits, fresh vegetables and vegetable dishes was insufficient. Eighty percent of students added extra salt to foods. More than 49% of students regularly consumed some forms of vitamin and/or mineral supplements. Thirty five percent of boys and 30% of girls smoked daily, 14.6% of boys and 5.9% of girls consumed alcohol at least once a week. Fifteen percent of boys and 11% of girls already tried taking drugs at least once. Only 50% of boys and 31% of girls were involved in a regular physical activity. Twenty two percent of fathers and 25% of mothers had university qualifications, whereas intermediate qualifications were recorded in 63 and 59%, lower qualifications occurred in 8 and 13% of parents, respectively. The incidence of smoking was 42% for fathers and 36% for mothers. Weekly alcohol consumption was recorded in 11%, daily consumption in 7% among fathers, the same figures for mothers were 4-4%, respectively. The incidence of hypertension, myocardial infarction, diabetes mellitus, and stroke was 26; 3.4; 5.4 and 1.5% in fathers and 32.9; 4.2; 6.6 and 1.5% in mothers. CONCLUSIONS: The data collected in the complex nutritional hygienic survey revealed serious deficiencies in the nutritional and life style habits of secondary school students. These factors considerably contribute to the unfavorable statistics of nutrition related non infectious diseases in adulthood. PMID- 14564925 TI - [Metastatic breast cancer in the uterine cervix: two cases of a rare occurrence]. AB - Authors report on the recurrence of breast cancer to the uterine cervix in two patients. In the first patient the tumor recurrence was suspected by cytology 43 months after she was cured for breast cancer. An ultrasound examination revealed the recurrence in the second patient who have been cured for the breast cancer for 53 months. The final diagnoses was proved by cervical conisation in the first patient and by hysterectomy in the second one. Authors call attention for the possibility of the rare site of symptom free recurrence. PMID- 14564926 TI - [Pharmacologic therapy of bronchial asthma]. PMID- 14564927 TI - [Percutaneous vertebroplasty]. PMID- 14564928 TI - [Description of a case and its consequences]. PMID- 14564929 TI - An epidemic of scurvy in Afghanistan: assessment and response. AB - In March 2002, there were reports of a hemorrhagic fever outbreak in western Afghanistan. It was later confirmed that the hemorrhagic symptoms and increased mortality were actually due to scurvy. Most aid workers did not include scurvy in the initial differential diagnosis because it is uncommon throughout the world and has mainly been reported in refugee populations in recent times. A rapid assessment confirmed the cases clinically, estimated a prevalence rate of 6.3% (a severe public health problem), and determined that the attack rates peaked each year in January and February (the end of the winter). Many Afghans have limited dietary diversity due to isolated locations, lengthy winters, the continuing drought of the last four years, asset depletion, and loss of livelihood. After numerous food and fortification options to prevent future outbreaks had been considered, vitamin C tablet supplementation was selected because of the relatively rapid response time as compared with other prevention methods. A three month course of vitamin C tablets was distributed to 827 villages in at-risk areas. The tablets were acceptable and compliance was good. No cases of scurvy were reported for the winter of 2002-03. The case study from Afghanistan demonstrates that scurvy can occur in nonrefugee or nondisplaced populations; vitamin C supplementation can be an effective prevention strategy; there is an urgent need to develop field-friendly techniques to diagnose micronutrient deficiency diseases; food-security tools should be used to assess and predict risks of nutritional deficiencies; and the humanitarian community should address prevention of scurvy in outbreak-prone areas. PMID- 14564930 TI - Food patterns during an economic crisis among pregnant women in Purworejo District, Central Java, Indonesia. AB - A cross-sectional study was conducted between 1996 and 1998. Six 24-hour recalls were performed during the second trimester of pregnancy among 450 women in Purworejo District, Central Java, Indonesia. The objectives of the study were to assess the food intake and food pattern among pregnant women before and during the economic crisis. Before the crisis, rich women had the highest intakes of animal foods, fats and oils, and sugar. Food intake among the urban poor and the rural landless poor subgroups was influenced by the emerging economic crisis. Although the price of rice increased, the intake of rice also increased among all subgroups. Rural poor women with access to rice fields increased their intake of rice and decreased their intake of nonrice staple foods (p < .05). There were significant decreases in the consumption of chicken by rich women and rural poor women with access to rice fields (p < .05). Rice was a strongly inferior good and remained an important supplier of energy, protein, and carbohydrate. Nuts and pulses were important suppliers of calcium and iron, and vegetables were an important supplier of vitamin A. Rich women increased their intake of nuts and pulses, vegetables, fats and oils, and sugar when their intake of rice increased (p < .05). The food patterns were based on rice, nuts and pulses, and vegetables, i.e., plant food. All but the rich women decreased their intake of nutritious foods such as meat, chicken, and fruits. The intake of nuts and pulses and of vegetables increased, whereas the intake of cooking oil and sugar remained constant. PMID- 14564931 TI - Recent experience with fortification of foods and beverages with iron for the control of iron-deficiency anemia in Brazilian children. AB - Iron-deficiency-anemia affects 30% of the world population. Women of reproductive age and children are the most affected. Iron supplementation in the form of tablets and syrups has not been successful in developing countries, and iron deficiency is still the most important deficiency related to malnutrition. Iron deficiency anemia affects physical and cognitive development at an early age in children, often resulting in irreversible outcomes. Studies from the last two decades have shown that the prevalence of iron-deficiency anemia can be reduced given adequate investments and political will directed at iron fortification of foods and liquids. A successful low-cost iron-fortification program incorporates implementation and strategic use of communication for program inception where education is in the forefront. A review of the available reports from experience in Brazil with iron fortification of foods and liquids is presented. PMID- 14564932 TI - Nutrition challenges in the next decade. PMID- 14564933 TI - Prevention of chronic diseases: WHO global strategy on diet, physical activity and health. PMID- 14564934 TI - Moving forward with complementary feeding: indicators and research priorities. International Food Policy Research Institute (IFPRI) discussion paper 146 (April 2003). PMID- 14564935 TI - Response to "Malnutrition and dietary protein: evidence from China and from international comparisons". PMID- 14564936 TI - Eating your way to higher test scores. PMID- 14564937 TI - Introduction to the proceedings of the International Research on Infant Supplementation (IRIS) Initiative. AB - In 2001, students and professionals from 13 nations were hosted by UNICEF for the International Workshop on Multi-Micronutrient Deficiency Control in the Life Cycle in Lima, Peru, May 30-June 1, 2001. Workshop participants engaged in both narrow and broad discussions of ways to combat multiple micronutrient deficiencies in developing countries. Preliminary data from four common-protocol studies conducted in Peru, South Africa, Indonesia, and Vietnam were presented. Participants also discussed the immediate, preliminary, and interim issues that could guide both policy and planning of future studies of multiple micronutrient deficiency. Among the studies highlighted was the International Research on Infant Supplementation (IRIS) I trial. A review of IRIS I yielded some confirmation of the efficacy using a crushable "foodlet" (i.e., cross between food and tablet) as a supplementation vehicle, as well as concerns about potential adverse consequences of nutrient-nutrient and nutrient-nutriture interactions. Other plenary topics illustrated the practical matters of how the IRIS I logistics and operations were built, and several focused on how to best design follow-up research on infant supplementation. Finally, a series of working groups allowed for in-depth discussions on the topics of community and policy, monitoring and implementation, and research. Researchers continue to try to identify efficient and effective programs suited to the low-income settings in which infant multi-micronutrient malnutrition occurs. The papers in these proceedings elaborate on several aspects of the IRIS study, and they are published in the hope that their analysis by readers will produce wider dissemination of the details of this devastating problem. PMID- 14564938 TI - Adequate dosing of micronutrients for different age groups in the life cycle. AB - Many studies of micronutrient supplementation in developing countries have used single-nutrient supplements with either vitamins or minerals. However, people in these countries often suffer from multiple, rather than single, micronutrient deficiencies. The objective of this paper is to discuss the factors that go into determining the adequate dosing of vitamins and/or minerals for people of different ages. To elaborate on the adequacy of micronutrient doses in supplements, a model described by the US FNB was used, which calculates the difference between the mean observed intake for an individual and the estimated average requirement for a life stage and gender group. This model allows estimating the degree of confidence that a certain nutrient intake (from supplements and diet) is adequate. The US/Canadian DRI values have been used as the basis for these calculations, from which it can be concluded that a daily supplement of one RDA of each micronutrient is adequate to cover the personal requirements of all individuals in each respective age and gender group of the population, provided that 20 to 40% of an RDA is supplied by the diet--likely a realistic value for developing countries. DRI values vary significantly between different age groups, reflecting changing needs over a life cycle. With the objective of a supplement to be adequate and safe, the design of a one-for-all supplement covering all age groups is not realistic. Such a supplement would either underscore or surpass the required intake of some of the age groups. Additionally the dosage of certain micronutrients might exceed the upper level of intake for lower age groups. Therefore, it is suggested that three different supplements following the one RDA concept for all micronutrients be developed for research use in developing countries for the following age groups; 1 to 3 years, 4 to 13 years, and females > 14 years (excluding during pregnancy). PMID- 14564939 TI - The foodLET vehicle designed for and used in the IRIS I intervention. AB - At the behest of UNICEF, conceptual specifications from the International Workshop on Micronutrient Supplementation throughout the Life Cycle (held in Rio de Janeiro, Brazil, in November 1999) were developed for a chewable, flavored multiple-micronutrient vehicle that was a hybrid of a food and a tablet (i.e., "foodLET"). Two varieties of foodLET were created: one with a single infant toddler RDA for selected micronutrients, and the other with two RDAs for the same vitamins and minerals. They were shipped and stored in special, hermetic blister packs to protect the physical form from crumbling and to protect the vitamins from oxidative damage. In the actual experience of delivering more than 40,000 foodLETs in four sites in diverse settings, the ability of the product to live up to the ideal characteristics can be assessed. PMID- 14564940 TI - Bioavailability of iron, zinc, folate, and vitamin C in the IRIS multi micronutrient supplement: effect of combination with a milk-based cornstarch porridge. AB - The effect of combining a multi-micronutrient supplement with a milk-based cornstarch porridge on the bioavailability of iron, zinc, folate, and vitamin C was evaluated using the plasma curve response over time (8 hours) in healthy women. Three tests were carried out in a crossover design: S (multi-micronutrient supplement), MS (multi-micronutrient supplement plustest meal), and M (test meal). Relative bioavailability was determined as the percent ratio of the area under the curve (AUC) in MS corrected by M, and AUC in S. Compared to S, AUC in MS was smaller for iron (p < .05), for zinc (p < .01), and for folate (p < .05), but not different for vitamin C. Relative bioavailability was lower (p < .05) than 100% for iron (80%), zinc (70%), and folate (85%). The decrease in bioavailability of these nutrients when the multi-micronutrient supplement is combined with a milk-based cornstarch porridge is small. Therefore, the tested meal is a suitable vehicle for the multi-micronutrient supplement. PMID- 14564941 TI - IRIS. I: a FOODlet-based multiple-micronutrient intervention in 6- to 12-month old infants at high risk of micronutrient malnutrition in four contrasting populations: description of a multicenter field trial. AB - Infants in developing countries are at risk of concurrent micronutrient deficiencies, because the same causative factors may lead to deficiencies of different micronutrients. Inadequate dietary intake is considered one of the major causes of micronutrient deficiencies, especially among poor and underprivileged children in developing countries. Operational strategies and distribution systems are often duplicated when supplementation programs for single micronutrients are implemented at the same time. The International Research on Infant Supplementation (IRIS) trial was conducted in four distinct populations on three continents: Africa, Latin America, and Asia. The participating countries were South Africa, Peru, Vietnam, and Indonesia. The study had a randomized, doubleblind, placebo-controlled design. Each country aimed to enroll at least 70 infants per intervention group (65 + 5 anticipated dropouts). The micronutrient vehicle was in the form of a "foodlet" (food-like tablet) manufactured as chewable tablets, which were easy to break and dissolve, and which had the same taste, color, and flavor for all countries. Children were randomly assigned to one of four 6-month intervention groups: group 1 received a daily foodlet containing multiple micronutrients; group 2 received a daily placebo foodlet containing no micronutrients; group 3 received a weekly foodlet that contained multiple micronutrients (twice the dose of the daily foodlet) and placebo foodlets on the other days of the week; group 4 received a daily foodlet containing only 10 mg of elemental iron. The IRIS Trial aimed to examine the prevalence of multi-micronutrient deficiencies in 6- to 12-month-old infants from rural populations, and to examine the efficacy of multi-micronutrient supplementation in infants from the different countries included in the study. This paper describes the general methodology of the IRIS trial and the operational differences among the country sites. PMID- 14564942 TI - The evolving applications of spreads as a FOODlet for improving the diets of infants and young children. AB - Action is needed to reduce the burden of micronutrient malnutrition in developing countries, and because low-income populations are vulnerable to deficiencies of multiple micronutrients, we need to move beyond approaches that comprise only single nutrients. The normal and evolutionary manner in which to consume nutrients is in the context of foods, both snacks and meals. Spreads are high viscosity-fat products prepared by mixing dried powdered ingredients with a vegetable fat chosen for its viscosity. Spreads are not traditionally used for feeding infants or young children and were initially proposed as a way to treat children recovering from severe malnutrition. In preparation for the International Research Group on Infant Supplementation (IRIS) III intervention, a sequel to the IRIS I study (which was the focus of a workshop in Lima, Peru, from May 30-June 1, 2001), the feasibility of preparing a FOODlet for feeding infants and young children was explored. Within the spectrum of intervention tools for micronutrient supplementation, tablets are a pharmaceutical form, fortified spreads are a food, and sprinkles are an intermediate approach. The issues still to be discussed and resolved with regard to creating such a spread include its specific micronutrient formulation, the capacity of young children to consume the required amounts (from either the FOODlet alone or the FOODlet mixed with other foods), the iron content and overall antioxidant protection of the spread matrix and its vitamins, potential allergenicity of proteins, and the economic implications of using such a FOODlet in low-income societies. PMID- 14564943 TI - IRIS. III: proposal of a multicenter efficacy study using a high-energy, micronutrient-dense spread. AB - There is an urgent need for the development of a high-energy, micronutrient-dense food, such as a spread, for clinical and emergency nutrition. The spread to be used in the International Research on Infant Supplementation (IRIS) III trial will contain carotene-rich palm oil as a source of vitamin A and energy, and heme iron from slaughterhouse blood as an iron source. All other micronutrients shall come from a pre-mix. The product shall be designed for young children, in particular, but can be consumed by adults, as well. An efficacy study will be conducted for six months in children 6 to 12 months old. PMID- 14564944 TI - Multiple micronutrient deficiencies: future research needs. AB - There has been a rising current of calls for a moratorium on international nutritional research in favor of an investment in intervention programs, per se. The topic of multiple-micronutrient supplementation reviewed at the International Workshop on Multi-Micronutrient Deficiency Control in the Life Cycle (May 30-June 1, 2001) has confirmed once again, however, the intimate interaction between program development and a supporting agenda of applied research. The areas of research required to produce successful intervention programs include biologic availability, safety and efficacy, communications and behavior, effectiveness, cost-effectiveness (efficiency), and food and pharmaceutical technology. Attention to safety and surveillance for unintended adverse effects has acquired new relevance as we analyze the multi-center International Research on Infant Supplementation (IRIS) I studies. All professionals involved in research projects in this area must assure both the quality and reliability of investigations and adhere to the highest principles of ethical conduct of research in human studies. The fundamental principles of research design and hypothesis development, quality assurance, reliability of measurements, and sound and unbiased interpretation of findings apply to all experimental science, and must be guaranteed for this mission. Agencies, academic institutions, and industry alike must work to create a system in which researchers can uphold these standards, and realize at the same time that the area of multi-micronutrient supplementation in developing countries can be a fertile area for training future researchers. PMID- 14564946 TI - RN's comp. claim based on PTSD resulting from short staffing, etc. Case on point: Smith-Price v. Charter Pines Behavioral Ctr., 584 S.E.2d 881-NC. PMID- 14564945 TI - Amputee's 'other leg' is injured by nurses & amputated. PMID- 14564947 TI - AL: wrong epinephrine dose--cardiac arrest: Ct. emphasizes the '5 Rs' of drug administration. PMID- 14564948 TI - LA: RN fails to prove disease was work related: expert's literature contradicts his testimony. PMID- 14564949 TI - Union nurse alleges sex discrimination in wrongful termination. Case on point: LaFournaise v. Montana Developmental Center, 2003 WL 22120720 P.2d-MT. PMID- 14564950 TI - [Role and part played by children's medical institutions in the medical support of training of youth for military service (Report II)]. PMID- 14564951 TI - [Experience with passenger motor ship re-equipment used for evacuation of the wounded and sick]. AB - In September 2002 the mobilization headquarters training was conducted on the base of Volga-Baltic steamship. During this training the passenger motor-vessel (the project 301) was re-equipped into sanitary-and-transport ship. The sanitary treatment coast post and the elements of evacuation receiving-room were developed by the personnel on the shore near the mooring wall. On the 3rd day the ship's sanitary treatment post, medical department (60 beds) with the wards for psychic patients (4 critical patients/ward), post for nurse on duty, room of physician on duty, dressing room, drugstore, autoclave room, clinical laboratory and collective defense post were created in the re-equipped rooms of the ship. The training has confirmed the advantages of casualty and patient transportation using the inner water-ways over the other types of transport. The following defects should be noted: the season work of river transport; frequent discrepancy of river trend with evacuation ways; comparatively low rate of transportation; different types of river ships used for medical evacuation. PMID- 14564952 TI - [Estimating the treatment and diagnostic and surgical work of the medical service army unit]. AB - To improve the objective quantitative evaluation of treatment-and-diagnostic work in the force link of the medical service the complex analysis of surgical work was conducted in 22 medical subunits. The urgency and informativity of the three indices of treatment-and-diagnostic work are substantiated: the number of completed treatment cases in the force link (%), the average treatment duration in the force link (days), the number of complications (%). The methods of index calculation are presented. The indices proposed allow to evaluate the treatment and-diagnostic work in the force link of medical service much completely and to optimize the assessment of medical care quality at the pre-hospital stage. PMID- 14564953 TI - [Cases of complex endoprosthetic surgery of the hip joint]. AB - 140 complex endoprosthetics of the hip joint were performed in 124 patients treated in the traumatology and arthrology Center of the Buyrdenko Main Military Clinical Hospital during the period from 1988 to 2001. It constituted 14.3% of all patients with diseases and trauma sequelae of such localization treated in the Hospital. There were 6 main patient groups in whom the pathological changes were considerably expressed and required the special approaches to the operation, i.e. the primary simultaneous bilateral hip joint endoprosthetics. The patients' age was 17-65 (the mean age 48.7), while during the conventional primary endoprosthetics the mean age was 68. We have developed the method of hip joint endoprosthetics performed in two stages. (Patent No 2173108 RF). This method was applied in 16 patients (group 1-11 and group 2-5 patients). The post-operative follow-up period lasted 1-6 years. Excellent and good results were obtained in 14 (87.5%) patients that were evaluated according to Harris's method. We think that it is reasonable to consider the primary complex endoprosthetics as the independent type of surgical intervention as it requires the special approaches to planning and performance of the operation, high skill and special up-to-date equipment. PMID- 14564954 TI - [Autonomic nervous system state in battle "mild" craniocerebral injury sequelae]. PMID- 14564955 TI - [Epilepsy pharmacotherapy: basic principles and tenets]. PMID- 14564956 TI - [Efficacy of antioxidants in the prophylaxis of respiratory diseases in young servicemen]. PMID- 14564957 TI - [Optimization of magnetic laser therapy in the treatment of male infertility]. PMID- 14564958 TI - [Legal aspects of government sanitary and epidemiologic surveillance of the armed forces of the Russian Federation]. PMID- 14564959 TI - [The surgeon L. A. Bekkers--a student of I. I. Pirogov]. PMID- 14564960 TI - [The Ocean Military Sanatorium--70 years]. PMID- 14564961 TI - [Department of Anesthesiology and Resuscitation of the Military Medical Academy- 45 years]. PMID- 14564962 TI - Leaders must act to avert crisis in older-adult services. PMID- 14564963 TI - Responding to an emergency for children's services. Innovative program seeks cure for over-reliance on crisis care. PMID- 14564964 TI - Is all the evidence in? Range of popular treatments subsist despite lack of science base. Is that damaging? PMID- 14564967 TI - Change principles applicable to variety of service settings. PMID- 14564966 TI - Lesson for providers: don't think about what you have, but what clients need. Client-focused mindset accounts for readiness to change. PMID- 14564968 TI - Facilitating change in the non-voluntary client. PMID- 14564969 TI - Differential diagnosis in MH settings: heed signs of substance abuse. PMID- 14564970 TI - Practice standards critical to minimizing risk associated with suicide. PMID- 14564971 TI - Ohio director leaves at 'top of her game'. PMID- 14564972 TI - Coronary heart disease in women. Past gaps and current understanding. PMID- 14564973 TI - Sex, survival bias, and mortality following acute myocardial infarction. AB - Several though not all studies have found an up to 2-fold increase in 30-day mortality in women compared to men admitted to hospital for ST-elevation myocardial infarction (MI), even after adjustment for baseline variables. These data however do not take into account the pre-hospital period. Indeed, three large WHO MONICA reports that included out-of-hospital events found no significant gender difference in overall 28-day mortality from MI, with more men dying before reaching the hospital (presumably of ventricular tachyarrhythmias) and more women dying after hospital admission (presumably of heart failure). Women compared to men exhibit enhanced vagal activity, both under basal conditions and during angioplasty-induced coronary occlusion, and this may afford protection against malignant ventricular arrhythmias. Epidemiological data indicate that women dying of ischemic heart disease are less prone to sudden death than men. Taken together, the above findings suggest that, following acute MI, significant gender differences lie not so much in overall mortality, but in the timing and mechanisms of death. PMID- 14564974 TI - Comparison of the outcome in men and women with chronic heart failure. AB - Recent studies have suggested some gender-related differences in the incidence, quality of care, response to therapy, and outcome in heart failure patients. The majority of studies have proposed a better survival for women compared to men. However, the under-representation of women in clinical trials leads to some uncertainty regarding the survival benefit. Some data suggest the possibility that ACE-inhibitors may be less beneficial in women as compared to men. Beta blockers seem to be effective both in women and men. Further studies are needed to clarify whether these differences may have a pathophysiological basis. PMID- 14564975 TI - Gender differences in the outcome of noninvasive cardiovascular treatment. AB - Cardiovascular diseases are equally common in both sexes with differences in pathogenesis, clinical presentation and outcome. The incidence of cardiovascular diseases progressively increases in women after the menopause and their development is related to risk factors and their interaction with the female hormones. The prognosis after myocardial infarction is worse in women because of a more difficult diagnosis (atypical symptoms, unclear ECG abnormalities) and because of social and economic factors. The role of hormone replacement therapy in atherosclerosis prevention is not yet clear, and there are many doubts about its administration because of the increased risk of breast cancer and cardiovascular events. Our best weapons against cardiovascular diseases are primary prevention and pharmacological therapy. The biological and pathophysiological mechanisms related to estrogen deficiency that may lead to the development of atherosclerosis are still unknown. Therefore, it would be useful to investigate the pathophysiology of atherosclerosis in women in order to improve primary prevention and the diagnostic and therapeutic strategies. PMID- 14564976 TI - Gender differences in diagnostic procedures. AB - This review examines the contribution of the literature to the controversial issue of diagnostic procedures in women affected by coronary artery disease (CAD), on which a large number of papers have been published. It has been reported that cerebro- and cardiovascular diseases represent the first cause of death in the New as well as in the Old World, Italy included. Some studies are conditioned by bias; one of these is the Framingham study, in which angina was reported and defined only clinically and for a relatively young age range, as a benign condition in women. Angiographic studies, such as the CASS, considered a super elected group of women referred to the hemodynamic laboratory for chest pain, which in the female gender often has atypical characteristics. In our opinion, it is mandatory to take into account: 1) what chest pain really means in women; 2) the fact that there are gender differences: women have a different biological and hormonal status, lifestyle, and perception of the disease; 3) that there is a different approach of the physicians to a woman with possible or suspected CAD. We suggest, therefore, a more peculiar and individualized diagnostic approach to women suspected as having CAD. This approach should also take the pre-test probability of disease into consideration. The first investigational step we recommend is the exercise ECG test; should this be unfeasible or not interpretable, an imaging and/or pharmacological stress test is advisable. In case of positive first test results, coronary angiography should be performed. PMID- 14564977 TI - Gender differences in the outcome of interventional cardiac procedures. AB - Prior studies have reported significant gender differences in the procedural outcomes after elective percutaneous transluminal coronary angioplasty (PTCA). Many of these differences have been explained by the presence of more comorbidities and worse clinical characteristics such as older age, unstable angina, congestive heart failure, diabetes mellitus, and hypertension in women than in men. Moreover, women have a smaller vessel diameter, more coronary tortuosity and different plaque composition compared to men that can lead to a higher dissection rate and a greater number of procedural complications. Although early data on PTCA suggested worse immediate results in women than in men, more recent data suggest that this difference is less marked. The introduction of stents with a low profile and a higher tractability and pushability has allowed the extensive application of these devices even in small and tortuous vessels improving the outcome of PTCA. This improvement has been higher in women than in men leading to the equalization of the immediate outcome in the two sexes, even if the baseline characteristics remain worse in women. In particular, mortality and the need for urgent surgical revascularization have become extremely low without any differences between sexes. However, some authors have still found a higher incidence of complications in the first period after the procedure due to stent thrombosis in the stenting era. For this reason, meticulous antiplatelet treatment should be prescribed and drugs such as glycoprotein IIb/IIIa inhibitors may also be considered advisable to reduce the excess risk in the female population particularly in women with prothrombotic risk factors such as diabetes. At 6 and 12 months similar rates of death, late myocardial infarction, and repeated revascularization have been shown in the two sexes. Coronary stenting and the use of glycoprotein IIb/IIIa inhibitors have also improved the immediate results in patients with acute myocardial infarction (AMI) undergoing primary PTCA. Studies comparing the outcome differences between women and men with AMI and treated with primary PTCA are limited but all suggest that women benefit more than men from this procedure. The in-hospital mortality in patients with AMI is significantly higher in the female than in the male population with a higher incidence of intracranial hemorrhage in women among tissue-type plasminogen activator-treated patients. Vice versa, women and men have a similar or a slightly higher in-hospital mortality after primary PTCA without intracranial bleeding complications. For this reason, an earlier diagnosis of AMI, an earlier hospital admission and an earlier primary PTCA should be the aims of management in order to improve the outcome in women with AMI and to equalize the procedural results in the two sexes. PMID- 14564978 TI - Treatment of hypertension in the elderly: new findings in older subjects. PMID- 14564979 TI - Variables correlated with early relapses after external electrical cardioversion of persistent atrial fibrillation. AB - BACKGROUND: The aim of this study was to identify the clinical and echocardiographic variables possibly correlated with the early relapses of atrial fibrillation (AF) after external electrical cardioversion (EC) in a large cohort of patients with persistent AF. METHODS: Two hundred patients (117 males, 83 females, mean age 67.9 +/- 8.7 years) with successful EC of persistent AF (> 72 hours) were included in the present study. In order to identify the predictors of early relapses (within 7 days) of AF, 16 clinical and echocardiographic variables were compared at univariate analysis. The variables with a p value < 0.10 at univariate analysis were subsequently analyzed at multivariate analysis. RESULTS: Seventy-five patients (37.5%) had relapses of AF within 7 days of EC. By univariate analysis only a younger age (65.9 +/- 8.9 vs 69.0 +/- 8.3 years, p = 0.01) was found to be significantly correlated with a higher incidence of early relapses of AF. At multivariate analysis no variable was found to be significantly correlated with early relapses of AF. CONCLUSIONS: In patients with persistent AF, recurrences of this arrhythmia within 7 days of EC occur frequently (37.5%). Multivariate analysis did not reveal any clinical or echocardiographic variable significantly correlated with the early recurrence of AF. PMID- 14564980 TI - Morbidity and mortality in 229 elderly patients with nonrheumatic atrial fibrillation. A five-year follow-up. AB - BACKGROUND: In the elderly the impact of atrial fibrillation on mortality and morbidity is substantial. Oral anticoagulant therapy reduces the risk of stroke by 70%; nevertheless, it remains largely underused. We evaluated, in a community prospective study, the factors associated with embolic events and death and the feasibility of oral anticoagulant therapy managed by general practitioners. METHODS: We enrolled at the Trieste Cardiovascular Center 229 patients aged > or = 65 years with non-rheumatic atrial fibrillation. At baseline, each patient underwent a transesophageal echocardiography and received instructions about oral anticoagulation. Patients were regularly followed by their general practitioner and finally evaluated at the Center. RESULTS: At baseline, the mean age was 73 years, 14% of patients were free of heart disease, 27% had had a previous embolic event, and 33% had an atrial thrombus. After a 5-year follow-up, 85% of the patients had been admitted to hospital, 17% had suffered an embolic event, and 35% were dead. Diabetes and the presence of a low flow in the left atrial appendage were predictive of embolic events. Heart failure, spontaneous echocontrast and aortic plaques were predictive of death. Anticoagulant therapy increased from 14 to 34% but the incidence of major bleeding did not change. The patients on anticoagulant therapy at follow-up constituted the group with the worst cardiovascular profile and embolic rate, but had a lower death rate (19%) compared with those on antiplatelet therapy (32%) and with those without antithrombotic therapy (67%). The hospitalization rates were respectively 78, 83 and 100%. CONCLUSIONS: In a group of elderly patients followed by their general practitioner with the support of a specialized cardiologic unit, oral anticoagulant therapy was well tolerated and associated with a significant decrease in mortality and hospitalization. PMID- 14564981 TI - Initial experience with a new on-line transthoracic three-dimensional technique: assessment of feasibility and of diagnostic potential. AB - BACKGROUND: Despite its wide diagnostic potential, three-dimensional (3D) echocardiography is a quite rarely employed technique. The ideal method to obtain transthoracic 3D imaging is on-line 3D echocardiography, but first-generation real-time instruments had technical limitations. A new on-line 3D technology which allows true real-time volume rendering of the cardiac anatomy has been recently introduced and its feasibility and diagnostic advantages have been evaluated in the clinical setting. METHODS: The system utilizes a "matrix" transducer with a dedicated software. It allows instantaneous acquisition and rendering on-line 3D images and interactive manipulation of 3D data. Eighty-three adult patients with various cardiac pathologies underwent on-line 3D echocardiography. Long- and short-axis views of the aorta, mitral valve and left ventricle and surgical views of these structures were attempted. The duration of acquisition and reconstruction, and the quality and incremental clinical value of 3D images in comparison with two-dimensional imaging were annotated. RESULTS: The mean time of 3D examination was 10 +/- 5 min; the mean number of acquisitions was 10.8 per patient. The quality of the 3D images was optimal in 39%, good in 37%, sufficient in 19%, and insufficient in 5% of the patients. In all cases at least one optimal or good live 3D image was obtained from the parasternal and apical views. The reconstruction of surgical or en face views was easily and rapidly (1 2 min) achieved by two experts in 3D echocardiography. The additional clinical values of 3D vs two-dimensional imaging was demonstrated in 7 patients with mitral valve disease, 3 with aortic valve pathology, and 3 with congenital heart disease. Several on-line 3D images that have not correspondence with two dimensional echocardiography were reconstructed, creating projections dedicated to the diagnostic goal. CONCLUSIONS: On-line 3D echocardiography can be easily performed in adult patients and allows for unique planes and projections. The instant rendering of 3D images facilitates the recognition of cardiac structures and increases the diagnostic potential of transthoracic echocardiography. PMID- 14564982 TI - In industrialized nations, a low socioeconomic status represents an independent predictor of mortality in patients with acute myocardial infarction. AB - BACKGROUND: It has been widely demonstrated that in the general population a low socioeconomic status (SES) represents a risk factor for coronary heart disease (CHD). Both the incidence of and the mortality due to CHD are significantly higher in subjects with a low SES. Conventional risk factors only partly account for the social gradient in CHD. The aim of this study was to evaluate whether mortality was associated with SES in patients with acute myocardial infarction (AMI). METHODS: To this purpose the articles dealing with AMI and SES, having a prospective cohort design, total mortality or sudden death as primary endpoint and multivariate analysis as statistical approach, were analyzed. RESULTS: Among the patients hospitalized for AMI, a low SES was associated with more advanced disease, as shown by a significantly higher prevalence of heart failure and of recurrent ischemic events. The in-hospital treatment and prescriptions at discharge were similar among the various socioeconomic groups. The pre hospitalization, in-hospital and long-term mortality were significantly higher in the most deprived subjects and a low SES represented an independent predictor (adjusted odds ratio ranging from 1.4 to 3.1). The few available data suggest the socioeconomic gradient as being a predictor even of sudden death. CONCLUSIONS: A low SES represents an independent predictor of mortality in patients with AMI. The current prognostic stratification of AMI patients is rather disappointing. In order to get a more precise prognostic stratification and to approach an adequate prophylaxis, a low SES should be utilized; however, the mechanisms mediating the adverse outcome of low socioeconomic groups should be further investigated. PMID- 14564983 TI - Late-onset Blalock-Taussig shunt occlusion due to a subclavian artery pseudoaneurysm. AB - A 3-month-old infant with tetralogy of Fallot presented with progressive severe cyanosis and intractable acidosis about 2 months after a successful modified right-sided Blalock-Taussig shunt. At cardiac catheterization, the suspected shunt malfunction was confirmed. It was due to a bulky, pear-like mass arising from the right subclavian artery and compressing the polytetrafluoroethylene conduit. Any attempt to recanalize the shunt by percutaneous techniques proved unsuccessful. At surgery, a huge dilation of the anterior wall of the right subclavian artery, that sharply bent the prosthetic conduit, was found. Pathologic examination revealed that the compressing mass was pseudo-aneurysmal in nature. Despite a second successful shunt operation with a dramatic clinical improvement, the patient died due to multiorgan failure 72 hours following surgery. Extrinsic compression by a false aneurysm is a rare cause of shunt occlusion that should always be suspected in patients presenting with a rapidly progressive shunt malfunction late after a successful shunt procedure. PMID- 14564984 TI - Asymptomatic malposition of a pacing lead in the left ventricle: the case of a woman untreated with anticoagulant therapy for eight years. AB - We report the case of a woman with a low-rate atrial fibrillation and a wire lead inadvertently inserted in the left ventricle through an ostium secundum defect. The malposition of the lead was diagnosed 8 years after the procedure on the basis of the presence of a right bundle branch block pattern of the paced QRS, at echocardiography and at chest X-ray. The patient, who was neither on antiaggregant nor anticoagulant therapy, was asymptomatic. Biventricular pacing is a useful tool in the therapy of patients with severe chronic heart failure and intraventricular conduction delay but the insertion of the lead in the left ventricle through the coronary sinus is not always feasible. Because a patent foramen ovale is a frequent occurrence in the adult population, we suggest that in case of an unsuccessful catheterization of the coronary sinus, the physician should search for a patent foramen ovale through which to introduce the wire lead into the left ventricle, especially if the patient is already receiving anticoagulant therapy. PMID- 14564985 TI - Thoracic epidural anesthesia for off-pump coronary artery bypass grafting in a spontaneously breathing conscious patient. AB - Minimally invasive coronary artery surgery is nowadays considered a safe surgical strategy. We report a case of a 70-year-old man with a high operative risk who underwent off-pump coronary artery bypass grafting using only high thoracic epidural anesthesia without intubation. The patient well tolerated each phase of the operation. The effect of the movement of the chest wall and the heart during spontaneous respiration did not influence the conduction of the operation. The postoperative course of the patient was uneventful. At follow-up the patient was asymptomatic. Repeat coronary arteriography and echocardiography respectively confirmed the patency of the anastomoses and the improvement of the left ventricular performance. In carefully selected cases thoracic epidural anesthesia for off-pump coronary artery bypass grafting in spontaneously breathing conscious patients is feasible and may offer many benefits. PMID- 14564986 TI - Acute severe coronary spasm associated with initial first dose of 5-fluorouracil chemotherapy. AB - Among the various pathophysiologic mechanisms proposed to explain the 5 fluorouracil cardiotoxicity, coronary vasospasm, occurring most frequently after the completion of the second or third dose of the cycle, has gained wide acceptance. We describe what to our knowledge is the first observation of typical Prinzmetal variant angina occurring very early after having started a 5 fluorouracil infusion administered as a chemotherapy regimen to a 66-year-old man with an adenocarcinoma of the right colon. PMID- 14564987 TI - A case of hypereosinophilic cardiomyopathy: additional value of the myocardial contrast agent SonoVue for the differential diagnosis of a cardiac mass. AB - We describe the case of a 37-year-old male referred because of hypereosinophilia associated with dyspnea. Transthoracic harmonic echocardiography showed an extensive myocardial infiltration and highlighted an intraventricular "in plus" image, whose characteristics were compatible with a diagnosis of intracardiac thrombus. The use of the myocardial contrast agent SonoVue (1 ml in bolus i.v. and 4 ml at an infusion velocity of 2 ml/min) allowed us to immediately identify, during left ventricular chamber opacification, the exact endocardial border of the left ventricular cavity and, later (when the residual SonoVue was evident only at the level of the myocardial walls), the true characteristics of the "in plus" image. This approach revealed the infiltration of the myocardial tissue and of both papillary muscles and chordae tendinae. The use of the myocardial contrast agent SonoVue may be, therefore, useful to distinguish the origin of "in plus" images often evident at echocardiography in the hypereosinophilic syndrome. PMID- 14564988 TI - Images in cardiovascular medicine. Sixteen-row multislice computed tomography of tuberculous pericardial abscess. PMID- 14564989 TI - Verbal integration: the interaction of participant roles and sentential argument structure. AB - This paper explores the interaction between sentence level syntactic information and the semantic information that is carried with the verb during sentence comprehension. A cross-modal integration task was employed to examine whether the number of participant roles (thematic roles associated with the central meaning of the verb) causes an increase in processing load during integration of the verb into on-going sentence comprehension. The effect of preceding sentential structural information (varied with respect to the number of argument and/or adjunct NPs preceding the verb) was also manipulated. Independent of the structural information preceding the verb, verbs with two participant roles were integrated into the sentence faster than verbs with three participant roles. This finding suggests that participant roles are stored with the representation of the verb and made immediately available during integration and comprehension. In addition, the syntactic distinction between arguments and adjuncts is also shown to play an immediate role in parsing and integration of language on-line. PMID- 14564990 TI - Two predictions of a compound cue model of priming. AB - This paper examines two predictions of the compound cue model of priming (Ratcliff & McKoon, 1988). Although this model has been used to provide an account of a wide range of priming effects, it may not actually predict priming in these or other circumstances. To predict priming effects, the compound cue model relies on an assumption that all items have the same number of associates. This assumption may be true in only a restricted number of cases. This paper demonstrates that when this assumption does not hold, the model does not easily predict priming. Second, the model fails on its own grounds in that it makes explicit predictions with respect to repetition priming effects, which do not match previously observed properties of repetition priming. PMID- 14564991 TI - The relation between coordinated interpersonal timing and maternal sensitivity in four-month-old infants. AB - The relation between mother-infant coordinated interpersonal timing, an automated microanalytic measure of dyadic vocal coordination, and maternal sensitivity was explored. Thirty-five mothers and their developmentally normal 4-month-old infants were audio-recorded during a 20-min laboratory vocal interaction session, that was later analyzed for degree of vocal coordination. Maternal Sensitivity ratings (Ainsworth & Bell, 1969) were based on a video-taped 45-min unstructured laboratory interaction period. A significant curvilinear relation between the degree to which mother coordinated her noninterruptive co-occurring speech to that of her infant was found and revealed that mothers highest in sensitivity were characterized by moderate levels of coordination. Examining mother-infant interaction at the specific behavioral level, while incorporating tests of nonlinear trends, may provide important information about the nature of sensitive parenting. PMID- 14564992 TI - Age preservation of the syntactic processor in production. AB - Two experiments are reported on the influence of cognitive aging on grammatical choice in language production. In both experiments, participants from two age groups (young and old) produced sentences in a formulation task (V. Ferreira, 1996) that contrasted conditions allowing a choice between alternative sentence arrangements (i.e., double object or prepositional dative) or that permitted no choice (i.e., prepositional dative only). Experiment 1 showed that older adults were able to formulate the alternative sentence arrangements with the same speed and fluency as young adults. Experiment 2 showed that cueing attention to one of the two object nouns to be included in the sentence resulted in the earlier expression of the cued noun in choice conditions, but with little evidence of a response time or dysfluency cost in the no-choice condition. As in Experiment 1, there were no substantive age differences in latencies or dysfluencies. These results support existing models for the mechanisms that choose between grammatical alternatives and bind phrases to available argument positions and provide evidence that older adults are not impaired in their use of these mechanisms. PMID- 14564993 TI - The use of the matrix method for the study of human motion: theory and applications. AB - Kinematics has been successfully used to describe body motion without reference to the kinetics (or forces causing the motion). In this article, both the theory and applications of the matrix method are provided to describe complex human motion. After the definition of a Cartesian coordinate frame is introduced, the description of transformations between multiple coordinate frames is given; the decomposition of a transformation matrix into anatomical joint motion parameters (e.g. Euler angles) is then explained. The advantages of the matrix method are illustrated by three examples related to biomechanical studies. The first describes a reaching and grasping task in which matrix transformations are applied to position the hand with respect to an object during grasping. The second example demonstrates the utility of the matrix method in revealing the coupling motion of the wrist between flexion-extension and radial-ulnar deviation. The last example highlights the indispensable use of the matrix method for the study of knee biomechanics, including the description of knee joint kinematics during functional activities and determination of in-situ ligament forces using robotic technology, which has advanced our understanding of the functions of the cruciate ligaments to knee joint kinematics. It is hoped that the theoretical development and biomechanical application examples will help the readers apply the matrix method to research problems related to human motion. PMID- 14564994 TI - [Research in synthesis of bioactive peptide RGD and the method for its grafting on PET surface]. AB - The aim of this experiment is to graft synthesizing Arg-Gly-Asp peptides (RGD) on the surface of polymer materials, combine endothelial cells with its special site, enhance the adhesion of endothelial cells on the surface, promote the blood compatibility of the biomaterials. Carboxy group (-COOH) was grafted on the materials surface by ultraviolet (UV) radiation, and the RGD serial obtained by liquid phase synthesis was successfully grafted on the disposed materials by chemical reaction. The endothelialization experiment was made also. The grafting results were measured by X-ray photoelectron spectroscopy (XPS), and endothelialization was observed using optical microscope and scanning electron microscope (SEM). The results indicated that the method improves the effect of materials endothelialization. The experiment has made successful use of UV grafting and chemical coupling methods to graft bioactive RGD onto PET film surface. This is a new method of grafting bioactive peptide. PMID- 14564995 TI - [Study on cyto-compatibility of PGLA film for periodontal guided tissue regeneration]. AB - This study was aimed to evaluate the cyto-compatibility of PGLA film for periodontal guided tissue regeneration (GTR) and their degradable products. Different extraction temperature, time and ratio were used to assess the cell relative growth rate (RGR) for PGLA. The degradable solution were brought into contact with cultured cells in vitro to observe the effects of cytotoxicity at 2, 4, 6, 8 and 10 weeks. The results showed when the extraction ratio was 0.1 g/ml, the extraction time has no effects on cytotoxicity at 37 degrees C. When the ratio was 0.1 cm2/ml, slight cytotoxic reaction appeared with the increase of extraction temperature (50 degrees C or 70 degrees C). When the extraction ratio was 0.5 cm2/ml in contract with the degradable solution at 37 degrees C for 72 h, the cell growth rate decreased. When the ratio was 6 cm2/ml, the cytotoxicity existed in some degree even if the temperature was at 37 degrees C for 24 h. After 2-4 w, the degradable solution had no obvious toxic effects on cells. However, the RGR gradually decreased as the degradation period increased. In conclusion, the effects of the extraction temperature, time, ratio and the degradation products accumulating in solution on RGR may exist. PGLA film has a good cytocompatibility. PMID- 14564996 TI - [Synthesis, characterization and in vitro release of poly (succinimide-co-4 aminobutanoic acid) by acid-catalyzed polycondensation of L-aspartic acid and 4 aminobutanoic acid]. AB - For the purpose of increasing the hydrophilicity of poly aspartic acid, a series of polymer of L-aspartic acid and 4-aminobutanoic acid with different ratios (mol/mol) were prepared. The copolymers were characterized by 13CNMR, DSC and x ray. The confirmed the structures of the polymers. In-vitro tests of release at phosphate buffer saline, enzyme solution of trypsin and papain (37.0 degrees C, pH = 7.4) were carried out. The result indicated that the polymers could be degraded in some degree, and that 4-aminobutanoic acid segments accelerated the degradation rate of the polymers. Skin irritation test and systemic acute toxicity test were carried out, which showed that the polymer was a nontoxic biomedical material. PMID- 14564997 TI - [Preparation and biological evaluation of PLA/chitosan composite materials]. AB - Hypersusceptibility test, pyrogen test, cell cultivation, and toxicity examination were applied in the biological evaluation of the poly(lactic acid) (PLA)/chitosan composite materials. The results indicated that all the materials were negative, conforming to the ISO10993-1. The cell could grow well on the surface of the materials. So the PLA/chitosan composite materials have good biocompatibility and can be planted in the body as scaffolds. PMID- 14564998 TI - [Studies of poly(vinyl alcohol)/hydroxylapatite hydrogels compounds for cartilage implantation]. AB - The structures and properties of polyvinyl alcohol (PVA)/hydroxylapatite(HA) composites were investigated. The components and processing conditions were studied for preparation of optimum compound hydrogels with good lubricating feature and high bioactivity. The properties, such as stretch strength, compress strength, friction, compress stress relaxation of various compound hydrogels were compared. The micro-morphology of PVA hydrogels and PVA/HA hydrogels were investigated by use of SEM. It was found that the addition of HA could improve the lubricating feature and mechanical strength of hydrogels, and its compress stress relaxation properties were closer to those of natural cartilages. PMID- 14564999 TI - [Hydrolysis of poly(L-lactic acid) fibers and formation of low crystalline apatite on their surface by a biomimetic process]. AB - Low crystalline apatite coating was formed on the surface of biodegradable poly(L lactic acid) (PLLA) fibers by a biomimetic process, i.e., by immersing the fibers in a modified simulated body fluid (SBF) at 37 degrees C and pH 7.3 after hydrolysis of the fibers in water. The apatite was characterized by scanning electron microscopy with energy dispersive X-ray spectroscopy, X-ray photoelectron spectroscopy, X-ray diffraction and Fourier transform infrared spectroscopy. Results showed that the fiber hydrolysis could accelerate the apatite formation but had little effect on the chemical and crystalline structure of the apatite. The structure of the apatite coating formed by the biomimetic method was similar to that of apatite in the natural bone. The bone-like low crystalline apatite coating might exhibit enhanced osteo-conductivity when the PLLA fibers are applied in bone reconstruction biomaterials. PMID- 14565000 TI - [Synthesis and characterization of biomacromolecular conjugates for magnetic resonance imaging]. AB - A two-step synthesis method was used to synthesize a series of N-(2 hydroxypropyl) methacrylamide (HPMA) copolymer-Proxyl conjugates as a contrast agent for magnetic resonance imaging (MRI) of solid tumors. The conjugates were characterized by Proxyl content using UV spectroscopy, electron spin resonance (ESR) spectroscopy and size-exclusion chromatography profile using a Fast Protein Liquid Chromatography instrument. The results show that a series of HPMA copolymer-Proxyl conjugates with incremental increase in Proxyl content were successfully synthesized and characterized, coupled with the macromolecular nature of the conjugates indicate that the copolymers have potential in MRI of solid tumors by enhanced permeability and retention effect. PMID- 14565001 TI - [Radioactivity of phosphorus implanted TiNi alloy]. AB - Exposed to neutron flow, the phosphorus implanted TiNi alloy gets radioactive. This radioactive material is used in vascular stent for prevention and cure of restenosis. Phosphorus implantation is carried out in a plasma immerged ion implantation system, and the dose of phosphorus implantation is in the range of 2 10 x 10(17) cm-2. After ion implantation, the alloy is exposed to the slow neutron flow in a nuclear reactor, the dose of the slow neutron is 1.39-5.88 x 10(19) n/cm2. The radioactivity of the TiNi alloy was measured by liquid scintillation spectrometry and radio-chromic-film dosimetry. The result shows that whether the phosphorus is implanted or not, the TiNi alloy comes to be radioactive after exposure to neutron flow. Just after neutron irradiation, the radiation dose of phosphorus implanted TiNi alloy is about one hundred times higher than that of un-phosphorus implanted TiNi alloy. The radiation difference between phosphorus and un-phosphorus implanted alloy decreases as time elapses. Within three months after neutron irradiation, the average half-decay period of phosphorus implanted TiNi alloy is about 62 days. The radiation ray penetration of phosphorus implanted TiNi alloy is deeper than that of pure 32P; this is of benefit to making radiation uniformity between stent struts and reducing radiation grads beyond the edge of stent. PMID- 14565002 TI - [Biological assessment of a new synthetic macromolecular resin class of comfort denture adhesive]. AB - This article devotes itself to evaluating the biocompatibility of synthetic macromolecular resin class of Comfort denture adhesive, which was developed by the present authors. Acute toxicity, haemolysis, cytotoxicity, sensitization and oral mucous stimulation were tested with standard method (ISO7406-1997 and YY0268 1995). The results showed that no toxic effect was observed with in vivo tests and no cytotoxic effect was observed with in vitro MTT assay. Haemolysis rate of the material (2.95%) indicated good hemaocompatibility. No local mucous membrane irritation reaction was noted after the relevant tests. The developed Comfort denture adhesive exhibited good biocompatiblility. PMID- 14565003 TI - [Experimental studies on the poly-hydroxybutyrate membrane modified by gamma radiation and mixed with calcium sulfate]. AB - The objective of this study was to learn the property of poly-hydroxybutyrate membrane (PHBm) modified by gamma-radiation and mixture of calcium sulfate, and to explore the possibility of using modified PHBm for guided tissue regeneration (GTR). The PHB was treated by 5 KGy gamma-radiation and mixed with 1/10 calcium sulfate. The modified PHB membrane was prepared by solvent-casting techniques. The mechanical properties and molecular weight of the modified PHBm were tested. Degradability of the modified PHBm was analyzed in vitro in a buffer solution of KH2PO4-Na2HPO4. Biodegradability and biocompatibility of the modified PHBm were inspected 1, 2, 3 and 6 months after the embedding of the modified PHBm into dogs. The morphology was analyzed by scanning electron microscopy (SEM) and molecular weight was tested to evaluate the biodegradability of PHBm. Biocompatibility of the modified PHBm was observed through tissue response by light microscopy. The extension strength and the extension strain at fracture of the modified PHBm were 23.8 MPa and 1.0% respectively. The morphologic observation of the modified PHBm at different terms showed that the modified PHBm was biodegraded gradually in vitro and in vivo. The capsule surrounding the modified PHBm was mainly composed of fibrocytes and few lymphocytes. The longer the time elapsed, the thinner the capsule enveloping the modified PHBm grew. The modified PHBm possesses satisfactory mechanical properties and biocompatibility, and it is biodegradable in vitro and in vivo. The modified PHB membrane could be applied as GTR membrane. PMID- 14565004 TI - [The effect of shear stress and flow pattern on proliferation of vascular endothelial cells]. AB - The non-random distribution of atherosclerosis was related to local hemodynamic environment. The stabilization of endothelium was important in this process. We studied the effect of magnitude of shear stress on proliferation of endothelial cells. It was shown the proliferation of endothelial cells was inhibited by shear stress, and was related to the magnitude of shear stress. A parallel plate sudden expansion flow chamber was constructed, and the effect of flow pattern was also studied. It was shown the inhibition effect produced by shear stress was decreased in this chamber. PMID- 14565005 TI - [Theoretical analysis and numerical simulation of effect of steel plate positions on steel plate rigidity in internal fixation of bone surgery]. AB - In this study we calculate theoretically and use FEM to simulate the effect of plate position relative to bending direction on the overall bending stiffness of the composite system plate-bone. The results show that for different bending directions the effect of the modulus of elasticity of the plate is negligible. Changing the position of a plate will often alter the stress obviously. During the operation, the steel plate should be assigned onto the tension side of the bone. PMID- 14565006 TI - [A system applying cyclic biaxial mechanical strain to cultured cells]. AB - To study the response of cultured cells to the cyclical biaxial biomechanical strain, a system was made according to the principle of Winston. The system was made of culture dish, pressure cavity and step motor. The culture dishes were made of polymethyl methacrylate and the bottom of the culture dishes were made of silicone rubber membrane. When the diameter of the silicone rubber tube and the distance between rollers and curvilinear plate changed, the pressure in the pressure cavity changed and the strain of the membrane changed from 0 to 4%. When the roll of the step motor changed, the frequency of the strain changed from 0.1 to 5 Hz. PMID- 14565008 TI - [Effects of tensile strain and loading time on the shape and cytoskeleton of the human periodontal ligament fibroblast measured by confocal laser scanning microscopy and immunity fluorescence technique]. AB - The periodontal ligament fibroblast (PDLF) was cultivated artificially as the cell to be tested, and then it was loaded with mechanical stress-strain of different values and for different times. The cell and nucleus projected areas and shapes as well as the structure of cytoskeleton were tested by use of confocal laser scanning microscope and immunity fluorescence technique. Then the relationship among the stress-strain, the time, the shape and the structure of cytoskeleton of the PDLF was detected. RESULTS: In the trial groups of 0, 8%, 12%, 16% strain values, the cell and nucleus projected areas were proportional to strain (stress) and time. The diameter, density and order of the structure of cytoskeleton increased in the strain and time dependent fashion. In the trial group of 20% strain values, the cell and nucleus projected areas decreased with the increase of time, and the structure of cytoskeleton became disorderly. It was demonstrated in this study that the shape and structure of cytoskeleton of PDLF underwent regular changes when the PDLF was loaded with the mechanical stress strain. PMID- 14565007 TI - [Experimental study on the viscoelastic properties of cancellous bone of the os calcaneus, os lunatum and os capitalum]. AB - We have researched the viscoelastic properties of the cancellous bones of the os calcaneus, os lunatum and os capitalum. The compressing stress relaxation experiment and the creep experiment in the vertical, horizontal and 45 degree directions on the os calcaneus were performed. The data and curve of the compressing stress relaxation and creep were obtained. By masing regression analysis we worked out the compressing reduced stress relaxation and creep functions and curves. The results show that the quantities of compressing stress relaxation and creep of the calcaneus in the vertical direction are larger than those in the other two directions. The initial quantities of creep of the os capitalum are larger than those of the os lunatum, and there are no significant different between the quantities of stress relaxation of the cancellous bones of the os lunatum and os capitalum. PMID- 14565009 TI - [Gene-enhanced tissue engineering: applications in osteoinduction using cultured mesenchymal stem cells transduced with the bFGF gene]. AB - To investigate the effect of basic fibroblast growth factor(bFGF) gene transfection on the proliferation and differentiation of mesenchymal stem cells (MSCs) and to provide basis for accelerating bone defect repairing using gene enhanced tissue engineering technology, Rabbit periosteum-derived MSCs were transfected with the full-length rat bFGF cDNA in vitro. The transient and stable gene expression of bFGF were determined by immunohistochemistry. The proliferation and the synthesis alkaline phosphatase (ALP) and osteocalcin(OC) of the transfected MSCs were also examined. The results showed that bFGF cDNA could be transferred into osteoblasts and expressed stably at least 4 weeks. The proliferation and OC content of genetically modified MSCs were increased significantly, whereas the ALP activity remained no change. In conclusion, transfer of gene encoding bFGF to MSCs increases its proliferation and osteogenesis property. Based on the successful conjunction of the existing techniques of tissue engineering with the novel possibilities offered by modern gene transfer technology, an innovative concept, molecular tissue engineering, was put forward for the first time. As a new branch of tissue engineering, it represents both a new area and an important trend in tissue engineering research. PMID- 14565010 TI - [Research in use of vascular endothelial cells to promote osteogenesis of marrow stromal cells]. AB - Osteogenesis deep in the materials is postponed because the blood vessels grow into the materials very slowly. It is one of the key challenges to the bone tissue engineering researchers. However, it was found that marrow stromal cells can produce vascular endothelial growth factor (VEGF) while endothelial cells produce bone morphogenetic protein (BMP). So we presume if we have co-culture of marrow stromal cells and endothelial cells on the biomaterials, osteogenesis and angiogenesis may be promoted simultaneously. In this experiment, we compare the alkaline phosphatase (ALP) activity and osteocalcin (OCN) production of three groups: the marrow stromal cells under activation of endothelial cells culture fluid, the induced marrow stromal cells, and the untreated marrow stromal cells. The results reveal that the ALP activity and OCN production of the marrow stromal cells under activation of endothelial cell culture fluid and the ALP activity and OCN production of the induced marrow stromal cells do not show statistically significant difference, but they are significantly higher than those of the untreated marrow stromal cells (P < 0.01). It is concluded that the VEGF in the endothelial cell culture fluid can promote osteogenesis as effective as osteogenic induction medium can do. PMID- 14565011 TI - [Airway beta-defensin-2 gene transfer enhances the bacterial clearing of rat lung]. AB - beta-defensins possess a broad spectrum of antimicrobial activity. In this study its in vivo antibacterial effect was evaluated by using gene transfer. Rat beta defensin-2 (rBD2) recombinant pBK-CMV-rBD2 and pCD-NA-3, 1-Myc-His(+)-rBD2 were constructed. Then, by use of liposome agent, the recombinants were delivered into rat airway via tracheal injection. The rBD-2 mRNA expression was detected in the trachea by RT-PCR and its protein expression was determined in the lungs by the tag His immunostaining, 24 hours after inoculation via trachea, the count of P. areuginosa in the lung of rat transfected with pBK-CMV-rBD2 markedly decreased, compared with the control (n = 8, P = 0.003). The data presented in this study provide evidence that airway beta-defensin-2-gene transfer can protect the rat against bacterial infection in vivo, suggesting the beta-defensins as part of the innate host defense system can be of potential applicability. PMID- 14565012 TI - [Experimental study and numerical calculation on cross-sectional temperature fields of animal's tongue]. AB - Bio-heat transfer in the tongue body is studied combining the mechanism of tongue inspection in Traditional Chinese Medicine. Parameters such as the temperature of lingual surface, the blood perfusion of the dog's tongue and so on are measured and the influences of the blood perfusion, the arterial blood temperature, the arterial cross-sectional areas and positions on the temperature distribution in the cross-sections are studied. Then the two-dimensional temperature fields in different cross-sections are numerically solved by the finite element method (FEM). The results show that the vascular cross-sectional areas vary with the elasticity change of the vessel walls resulting from the variation of the blood perfusion. And the temperature distribution in the cross-section mainly depends on the arterial cross-sectional areas and positions. The results can help to analyze the bio-heat transfer characteristic of the tongue. According to this method, the sectional temperature fields in whatever place of the tongue can be calculated under different blood perfusion and on the condition that the influence of the venous blood temperature on the temperature field of the tongue is considered. We can further probe into the relationship between the temperature fields of the lingual surface and that of cross-sections. This can provide the foundation for further investigation into the bio-heat transfer mechanism and the calculation on three-dimensional temperature fields of the tongue. PMID- 14565013 TI - [A microcomputerized pulsed water-sac massage with drug penetration instrument for treatment of male patients with immunity infertility accompanied by chronic seminal vesiculitis]. AB - This article describes the development of an innovative microcomputerized pulse water-sac massage with drug penetration instrument and reports its effectiveness in the patients with immunity infertility accompanied by chronic seminal vesiculitis. The instrument was developed on the basis of the pathological characteristics of immunity infertility accompanied by chronic seminal vesiculitis, and along the lines of our practice in integrating the modern theory of traditional Chinese medicine, with multiple techniques of biomedical engineering sciences. 181 male patients with immunity infertility accompanied by chronic seminal vesiculitis received the treatment. Of these cases, 135(74.6%) were cured, 37(20.4%) were treated with significant therapeutic effects and 9(5.0%) with improved effects. The results of antisperm antibody (AsAb) tests became negative in 85.6% of the patients after treatment, and the pregnancy rate of their wives was 49.1%. No mild adverse effects were observed in all cases. PMID- 14565014 TI - [A study on the preparing of magnetic doxorubicin nanoparticles and its heat effect under a magnetic field]. AB - The aim of this research report is to introduce a method of making Doxorubicin magnetic nanoparticles for tumor treatment and to evaluate its heat effect under 100 KHz magnetic field. The preparing of the nanoparticles was carried out by stirring with supersonic mixer and cold drying. The shape and diameter of the nanoparticles were observed by penetrating electron microscopy. The nanoparticles were spherical in shape, and most of them were 200 nm in diameter. The changes of the temperature under a magnetic field in agarose gel and distilled water were measured respectively. The temperature went up with the increase of the amount of nanoparticles and the magnetic field intensity. The temperature finally remained constant due to the balance of heat generation and its transfer to the surroundings. The heat transfer was faster in water in the agarose gel. PMID- 14565015 TI - [Effects of therapeutic dose on temperature rise induced by high intensity focused ultrasound in tissue]. AB - Based on the well-known Pennes' bioheat equation, we analyzed theoretically the temperature rise in tissue during high intensity focused ultrasound(HIFU) irradiation, and measured the focal temperature rise induced by HIFU in the treatment of freshly excised cow liver with different acoustic intensity and exposure time. The results showed that a threshold exposure time always existed under a certain acoustic intensity. Temperature rise was slow when exposure time exceeded the threshold exposure time. The greater the acoustic intensity was, the earlier the threshold time appeared. The focal temperature rise and the relative cumulative thermal dose (RCTD) increased with the increase of acoustic intensity and exposure time. For a certain therapeutic dose, the effects of acoustic intensity on focal temperature rise were more distinct than the effects of exposure time on focal temperature rise. Therefore, the optimal HIFU therapeutic dose should meet the need, i.e. moderate acoustic intensity, and the exposure time be the threshold exposure time under this acoustic intensity. PMID- 14565016 TI - [FHIT gene is abnormal in tongue carcinoma cell line]. AB - To study the alteration of FHIT gene in tongue carcinoma Tca8113 cell line, total RNA of Tca8113 cells was extracted. The transcript of the FHIT gene of the Tca8113 cell line was detected with nest RT-PCR, and DNA was sequenced. The result showed that abnormal transcript (about 247 bp) of FHIT gene was detected in the Tca8113 cell line. The sequence analysis of the aberrant cDNAs revealed deletions of exons 1-8. Therefore, the deletion of the FHIT gene in Tca8113 cell line might support the hypothesis that the FHIT gene alteration is involved in the development of tongue carcinoma. PMID- 14565017 TI - [Implementation of mutual information based medical image registration methods]. AB - Image registration methods based on mutual information, including mutual information and normalized mutual information, have been accepted as the most accurate and efficient methods. But there are many fluctuations in the registration functions that hinder the optimization procedure and lead to registration failure in intra-modal registration. We found that besides the interpolation artifacts, the uncertainty of the changing of entropy with the changing of overlap also contributes to the fluctuations. The effect of interpolation artifacts can be eliminated, but it is difficult to eliminate the effect of uncertainty of entropy. Luckily, this effect is not significant in normalized mutual information. Normalized mutual information is more stable and robust than standard mutual information and its better performance and wider application can be expected. PMID- 14565018 TI - [Eliminating artifacts of EEG data based on independent component analysis]. AB - As a new array processing technique, independent component analysis(ICA) is an effective means to resolve the blind source separation(BSS) problem. Based on the brief introductions of ICA theory and algorithm, we apply ICA to the removal of ocular artifacts from EEG recordings. The EEG data collected from the human scalp is actually the mixtures of some independent components. It is coincident with the basic assumptions of ICA. Compared with the traditional methods of artifacts elimination, ICA, a kind of spatial filter, is not restricted by the case of spectrum overlapping, and it has a good reservation of useful detail signals. In addition, the inverse weight matrix of ICA can be used to reflect the topographic structure of different independent sources of EEG. PMID- 14565019 TI - [Application of support vector machines to classification of blood cells]. AB - The support vector machine (SVM) is a new learning technique based on the statistical learning theory. It was originally developed for two-class classification. In this paper, the SVM approach is extended to multi-class classification problems, a hierarchical SVM is applied to classify blood cells in different maturation stages from bone marrow. Based on stepwise decomposition, a hierarchical clustering method is presented to construct the architecture of the hierarchical (tree-like) SVM, then the optimal control parameters of SVM are determined by some criterion for each discriminant step. To verify the performances of classifiers, the SVM method is compared with three classical classifiers using 3-fold cross validation. The preliminary results indicate that the proposed method avoids the curse of dimensionality and has greater generalization. Thus, the method can improve the classification correctness for blood cells from bone marrow. PMID- 14565020 TI - [Detection of surface EMG signal using active electrode]. AB - Research of surface electromyogram(EMG) signal is important in rehabilitation medicine, sport medicine and clinical diagnosis, accurate detection of signal is the base of quantitative analysis of surface EMG signal. In this article were discussed how to reduce possible noise in the detection of surface EMG. Considerations on the design of electrode unit were presented. Instrumentation amplifier AD620 was employed to design a bipolar active electrode for use in surface EMG detection. The experiments showed that active electrode could be used to improve signal/noise ratio, reduce noise and detect surface EMG signal effectively. PMID- 14565021 TI - [Frequency detection of the first heart sound based on wavelet transformation]. AB - According to the valvular theory, the vibrations affected by the mitral and tricuspid valves closure in early systole produce the first heart sound (S1). S1 usually includes many frequency components. In this paper, a method using the multi-resolution analysis of wavelet transformation is recommended for detecting the frequency range of S1. First, S1 was decomposed into different levels on frequency. Then the normalized Shannon energy of the different levels was calculated. The level containing the maximum energy is the major components' level of S1. The frequency range of this level is the major frequency range of S1. The frequency range of S1 was successfully detected by the method. PMID- 14565022 TI - [The effects of electrostimulation upon soldier's circadian of melatonin in plasma]. AB - Physiological and behavioral rhythms are governed by an endogenous circadian clock. In this paper are reported the studies on soldier's circadian of melatonin concentrations by means of electricity and light pulses presented to the popliteal region (behind the knee). The results showed that the phase of melatonin concentrations can be regulated by both the electricity pulses and the light pulses. A systematic relation was found between the timing of the electro stimulating and the magnitude and direction of phase shifts, resulting in the generation of a phase response curve. The phase response curve displayed photic model. These findings have implications for the development of more effective treatments for sleep and circadian rhythm disorders. PMID- 14565023 TI - [Enhancing the cytotoxicity of antitumor drugs through electromagnetic pulses]. AB - We chose Hela cells as research object and studied the cytotoxicity generated by cyclophosphamide, an antitumor drug, after cell electroporation by the use of electromagnetic pulses. Comparison between the electroporation group and the contrast group revealed the greatly enhanced cytotoxicity of the electroporation group, indicating that under some conditions electromagnetic pulses can enhance the cytotoxicity of antitumor drugs. The results of this study provide reliable evidences and a feasible approach for clinical treatment of tumor. PMID- 14565024 TI - [The complex impedance frequency response and the equivalent circuit model of human brain]. AB - The complex impedance measurements of human brain tissue in vitro are made by using four-electrode measurement methods in the frequency range from 0.1 Hz to 1 MHz. The Solartron 1255 B frequency response analyzer is used. The frequency response curves of the imaginary part of human brain's complex impedance do not appear as a single peak curve which other bio-tissues show, and the Cole-Cole diagram of human brain is different from the others as well. The construction of the equivalent circuit model obtained is more complex. However, these characteristics of human brain are all the same as rabbit brain's. The equivalent circuit model obtained will be helpful to constructing the equivalent circuit model of human head in the EIT researches. PMID- 14565025 TI - [Study on the function localization during different brain calculation tasks]. AB - The aim of this study is to define the anatomical localization of corresponding brain function area during calculating. The activating modes in brain during continuous silent calculating subtraction and repeated silent reading multiplication table were compared and investigated. Fourteen volunteers of right handedness were enrolled in this experiment. The quite different reaction models in brain area during the two models of calculation revealed that there are different processing pathways in brain during these two operating tasks. During continuous silent calculating, the function areas were found localized on the posterior portion of superior and middle gyrus of frontal lobe and the lobule of posterior parietal lobe (P < 0.01, T = 5.41), demonstrating that these function areas play an important role in the performance of calculation and working memory. During repeated silent reading multiplication table, the activating cluster was found only located in in the gyri occipital lateral(P < 0.01, T = 4.77). PMID- 14565026 TI - [Analyzing heart period signal for fetus and pregnant woman in perinatal period]. AB - Analyzing the digital characteristics (chaotic and spectral features) of fetal and pregnant woman's heart period signal (HPS) can assess the autonomic nervous system function. Extracting and analyzing HPS of fetal and pregnant woman in perinatal period were realized by using the method of visual programming. The subjects were in supine position. Electrocardiography-signal (ECS) from the leads placed at symphysis pubic bone to inferior border of abdominal wall was acquired. ECS was preprocessed by wavelet filter. The sophisticated technique developed by our laboratory was used to analyze the digital characteristics of HPS. The system could be used to assess fetal and pregnant woman's autonomic nervous system function, furthermore, sympathetic and parasympathetic nervous function could be evaluated respectively and quantificationally. The system also could be used to prognosticate fetal distress. The digital characteristics of fetal and other age groups' HPS, which vary with age, suggest the physiological process of development, mature and senility of autonomic nervous system; based on it, we could find the way of anti-senility. Some digital parameters of fetal HPS stand between those of newborn and adult, which may imply that fetal autonomic nervous regulation is influenced by mother's nerve and endocrine system. PMID- 14565028 TI - [Scientometrics and bibliometrics of biomedical engineering periodicals and papers]. AB - This investigation was made to reveal the current status, research trend and research level of biomedical engineering in Chinese mainland by means of scientometrics and to assess the quality of the four domestic publications by bibliometrics. We identified all articles of four related publications by searching Chinese and foreign databases from 1997 to 2001. All articles collected or cited by these databases were searched and statistically analyzed for finding out the relevant distributions, including databases, years, authors, institutions, subject headings and subheadings. The source of sustentation funds and the related articles were analyzed too. The results showed that two journals were cited by two foreign databases and five Chinese databases simultaneously. The output of Journal of Biomedical Engineering was the highest. Its quantity of original papers cited by EI, CA and the totality of papers sponsored by funds were higher than those of the others, but the quantity and percentage per year of biomedical articles cited by EI were decreased in all. Inland core authors and institutions had come into being in the field of biomedical engineering. Their research topics were mainly concentrated on ten subject headings which included biocompatible materials, computer-assisted signal processing, electrocardiography, computer-assisted image processing, biomechanics, algorithms, electroencephalography, automatic data processing, mechanical stress, hemodynamics, mathematical computing, microcomputers, theoretical models, etc. The main subheadings were concentrated on instrumentation, physiopathology, diagnosis, therapy, ultrasonography, physiology, analysis, surgery, pathology, method, etc. PMID- 14565027 TI - [Nonlinear analysis on the EEG information of rat epileptic model]. AB - The aim of this study was to develop a new method of epileptic prediction using nonlinear dynamic theory. When rat was falling sickness, its EEG was researched by using approximate entropy and correlation dimension. The results showed the approximate entropy and correlation dimension during epileptic seizure are obviously lower than those before seizure and after seizure. The span of time before seizure is a special phase. Before the seizure symptom appeared, the complexity of EEG had begun declining. Thus, the outbreak of epilepsy could be predicted in short time using nonlinear dynamic methods. PMID- 14565029 TI - [An improved PID algorithm for temperature control used by tumor combined therapeutic instrument]. AB - The development of a multi-functional tumor combined therapeutic instrument based on improved PID algorithm is introduced. It is based on the theory of 1 + 1 = 3 or 1 + 1 + 1 > 3 curative effect enhancement by tumor combined therapy, and it can be used solely, or with the combination of chemotherapy or actinotheraphy or both for tumor hyperthermia. By adopting improved PID temperature control algorithm, the problem of temperature control precision reduction due to process characteristics changing with different heaters was solved, thus ensuring homogeneous and smooth radio frequency heating by different heaters to intracavity tumor foci. These experiments demonstrated that the new algorithm has strong adaptability and anti-disturbance capability, the equipment works stably and reliably, and it can control therapeutic temperature precisely (+/- 0.1 degree C), which indicates a good value in clinical application. PMID- 14565030 TI - [A new method using the technique of cubic spline interpolation to remove baseline wander]. AB - The present methods available for correction of baseline wander with cubic spline interpolation technique all determine the fiducial points from P-R intervals. However, the determination of the fiducial points can be adversely affected by a noisy signal or an uneven P-R interval. In this paper, we introduce a new method of determining fiducial points by using the feature of high frequency of QRS wave. First, we correct the baseline approximately by using a FIR filter, then find the position of R peak and its amplitude change after filtering. The experiments demonstrate that the previous position and amplitude change can be looked as the position and amplitude of a fiducial point. Finally, we estimate a continuous baseline by joining fiducial points with spline. Our experiment results show that this is a better filtering method in comparison with the one introduced in reference paper [3]. PMID- 14565031 TI - [The design of a new respiratory detecting system using impedance method]. AB - A coupling principle of reflecting impedance based on resonance is designed to achieve impedance detection with high sensitivity. It is characterized by small impelled current, high sensitivity and simple circuit. The principle can be used not only in detecting human respiratory impedance, but also in detecting the bio impedance of other human organs. It may find wide application in this aspect. PMID- 14565032 TI - [Study on preparation of a wound healing agent: fibronectin]. AB - In search of the optimal preparation method for large-scale purification of human plasma fibronectin, we adopted affinity chromatography with gelatin and the Sepharose 4B activated with cyanogen bromide to purify fibronectin from type "C" plasma of healthy males, and scanned the best method under the conditions of different amount of plasma loading and different residence time in column. In a given column volume of gelatin, the absorbent was related with the plasma residence time in column and the total amount of plasma loaded. As a result, the optimal loading amount of plasma is 150 ml, and the residence time is 20 minutes. The preparation method, herein, has been proved to require small amount of plasma and yield large amount of fibronectin. PMID- 14565033 TI - [A new approach for improving antithrombogenicity in centrifugal pump]. AB - For long-term application of the rotary pumps, it is necessary to solve the problems of bearing wear and thrombosis along the bearing. Currently, many investigators choose the magnetic bearing to realize zero-friction and no contact between the rotor and stator; the former avoids the mechanical wear and the latter eliminates the possibility of thrombus formation. We tried and found that it is difficult to apply a magnetic bearing to rotary pump without disturbing its simplicity, reliability and implantable; therefore, we have developed a much simpler and much more creative approach to achieving the same results. Instead of the sliding bearing, a rolling bearing has been devised for the pump; its friction is about 1/15 of the sliding bearing. Furthermore, a wear-proof material of ultra-high-molecular weight polythene has been adopted in making the rollers, their anti-wear property in 8 times better than that of metal. Thereby, the service life of the bearing has extended to several years. For preventing the thrombus formation along the bearing, the impeller reciprocation axially as the impeller changes its rotating speed periodically to produce a pulsatile flow. The reciprocation is a result of the effects of a magnetic force between the motor rotor and stator, and a hydraulic force between the blood flow and the impeller. Similar to piston pump, the oscillating impeller can make the blood in and out of the bearing, resulting in wash-out once a circle. This is obviously beneficial to preventing thrombosis along the bearing and in the pump. The endurance tests with saline of this novel pump demonstrated a durability of the device. It promises to be able to assist the circulation of the patients permanently and to be able to replace the heart transplantation in the future. PMID- 14565034 TI - [Development of the vascular prosthesis research]. AB - The search for a nonthrombogenic material with the potential for use in small diameter vascular graft applications continues to be a field of extensive investigation. This article describes the choice of biomaterials used as vascular prosthesis, the innovation of construction of tissue-engineered blood vessels, the indispensability, methods and the effect produced by surface modification of vascular prosthesis. The article also points out that research achievements of vascular prosthesis must be made with the exploitation of new nonthrombogenic biomaterial and the development of tissue engineering. PMID- 14565035 TI - [Research and development of A-W bioactive glass ceramic]. AB - Studies on bioactive glass and glass-ceramic are important research high-lights in the field of biomedical materials. Due to their bioactivity, these materials can form a tight chemical bond with the living bone, when implanted. As a preeminent kind of these materials, A-W(Apatite/Wollastonite) bioactive glass ceramic has not only the excellent bioactivity and biocompatibility, but also the eminent mechanical properties, so it has been largely applied and developed in clinical practice. The development, preparation, properties, applications and the mechanism of its bond with bone are introduced in this paper. We will also put forward the prospect of the research and development of A-W bioactive glass ceramic. PMID- 14565036 TI - [The advance of bioactive peptide RGD in the research of bone regeneration]. AB - Clinically, there has been so far no effective way to repair the bone-missing of large extent due to gash, infection and removal of tumor. The solution of this problem can be assisted by the addition of bioactive substances to substrate materials, because the growth of peripheral tissue and the fiber tissue growing the materials can be induced to the direction of bone-tissue by these biomaterials with bioactive peptides. The peptide Arg-Gly-Asp is the point between the integrin which comes from membrane and the ligand. In certain cases, the artificially synthesized RGD can be competitively combined with the integrin on cell surface, and outer-cell information is transmitted into cells, which will cause a series of physiological changes in cells. Presently, it is reported that the RGD has the ability to induce the growth of osteoblasts, restrain the adhesion between osteoclasts and substrates. This paper reviews and introduces the progress made with the work of RGD-inducing bone regeneration. PMID- 14565037 TI - [Research development of hydroxyapatite-based composites used as hard tissue replacement]. AB - Hydroxyapatite has been considered as the most promising materials for hard tissue replacements, due to its similar chemical composition and crystallographic structure to that of bone mineral. But the brittleness is one of the most serious obstacles for its wider applications as load-bearing implants. Therefore, various HA composites get much attention. In the present paper, HA composites were introduced according to the kind of reinforcement. Although bioactive ceramics, bioactive glass or glass-ceramic, bio-inactive ceramics, polymers and metals all have been used to fabricate HA composites, no one can well satisfy the requirements for hard tissue replacement. The vital problem of the existing HA composites is that the biological properties cannot match with the mechanical properties well. PMID- 14565038 TI - [Morphological parameters for endothelial cells under shear stress]. AB - Shear stresses acting on vascular endothelial cells due to blood fluid flow play an vital role in regulating their morphology, structure, growth rate and functions. The change in morphology is an instinctive response to biomechanical environment and is an indicator for the functional changes in the cells. In this paper, the morphological research literatures of endothelial cells in the recent years are reviewed and the common morphological parameters are described in detail. At the end, the difficulties of quantitative study of endothelial cell morphology and the future direction of morphological parameters research are also discussed. PMID- 14565039 TI - [Introduction to medical data mining]. AB - Modern medicine generates a great deal of information stored in the medical database. Extracting useful knowledge and providing scientific decision-making for the diagnosis and treatment of disease from the database increasingly becomes necessary. Data mining in medicine can deal with this problem. It can also improve the management level of hospital information and promote the development of telemedicine and community medicine. Because the medical information is characteristic of redundancy, multi-attribution, incompletion and closely related with time, medical data mining differs from other one. In this paper we have discussed the key techniques of medical data mining involving pretreatment of medical data, fusion of different pattern and resource, fast and robust mining algorithms and reliability of mining results. The methods and applications of medical data mining based on computation intelligence such as artificial neural network, fuzzy system, evolutionary algorithms, rough set, and support vector machine have been introduced. The features and problems in data mining are summarized in the last section. PMID- 14565040 TI - [Advance in signal analysis of auditory evoked potential]. AB - Signal analysis of the auditory evoked potential (AEP) has been the interesting field in biomedicine and signal processing for a long time. Recent advances in the analytic method of AEP are summed up in this paper. Seven methods, including spectral analysis, adaptive filtering, wavelet transform, chaos theory, artificial neutral network, regularization, and independent component analysis are mainly introduced. All of them are promising for extracting the signal with less stimulation and adequate quality, and for displaying their efficient application in the analysis of AEP. PMID- 14565041 TI - [Record, analysis and application of EGG]. AB - Electrogastrography (EGG) is the cutaneous recording of the myoelectrical activity of the stomach using surface electrodes and it is more and more attractive due to its noninvasiveness. It has been used in medical research and clinical diagnosis. It is evident that with the use of this noninvasive measurements of gastric electrical activity(GEA) that we can diagnose gastric dysfunction. In this article, record, analysis, application and trend of EGG are reviewed. PMID- 14565042 TI - [Migraine in children]. PMID- 14565043 TI - [Selective neck dissection in the treatment of pN1/2 lymph node metastases in the neck]. AB - INTRODUCTION: As yet there is no single reliable and accurate method for detection of neck lymph node metastases. Therapeutic approaches differ from one author to other. The aim of this paper was to establish the extent to which, with good control of primary process, we can control spreading of malignant disease by means of selective neck dissection. MATERIAL AND METHOD: This retrospective study included 595 surgically treated patients in the period 1990-1998. There were 525 patients with malignant laryngeal tumors, and 70 patients with malignant hypopharyngeal tumors. Preoperative diagnostics of enlarged lymph nodes was based on palpation of the neck, without CT, US, NMR diagnostics. With all risky N0 patients, selective neck dissection was performed for presence of occult metastases. Intraoperative frozen section analysis was not performed. Adjuvant radiatitherapy was performed in all patients in whom presence of neck lymph node metastases was histologically proved. RESULTS: Selective lateral neck dissection was performed in 389 (65.4%) patients. In 78 (20%) patients, lymph node metastases were pathohistologically detected. In 5 (6.4%) transitional cellular cancer was histologically diagnosed, and the remaining 73 (93.6%) presented with squamous cell cancer. Postoperative radiation therapy was applied in 54 patients (69.2%) while 24 (30.8%) were not irradiated. 5-year survival was achieved in 18 (23.1%), and 3-year survival was achieved in 15 (19.2%) patients. Out of 45 patients who lived less than tree year, 18 (40%) presented with metastatic relapse and fatal outcome. Relapse of neck metastases appeared in 12 (11.9%) on the side and in the zones of lymph nodes which were included in neck dissection. Recurrence of neck metastases appeared in 8.3% of patients who were not irradiated postoperatively and in 32.1% of cases irradiated postoperatively. DISCUSSION: This study includes comparison of our results with results of literature data. CONCLUSION: Neck lymph node metastases point to advanced malignant process of the third or forth stage of the disease when results are the worst and 5-year survival decreases with or without adjuvant radiotherapy. PMID- 14565044 TI - Correlation between clinical symptoms and diagnosis of trichomoniasis in women. AB - The aim of this study was to investigate correlation between clinical symptoms and diagnosis of trichomoniasis in women. 200 women were included in the study. Swabs were taken from all patients from the posterior vaginal fornix. Each sample was examined using the following five methods: wet mount, Giemsa stain, acridine orange fluorescence stain, cultivation in Diamond medium and PCR method. Trichomoniasis was diagnosed in 27 women using any of the applied methods and 33.3% presented with typical frothy yellow-green discharge, characteristic for tichomoniasis and yellowish discharge characteristic for the third group of vaginal discharge. White discharge, characteristic for Candida infection, was found in 18.5% of patients with diagnosed trichomoniasis. Finally, 14.8% of positive patients had a normal discharge. Based on the results of our study we came to the conclusion that microbiological investigations are necessary for accurate diagnosis of trichomoniasis, as well as for revealing asymptomatic infections, in order to prevent spreading of this relatively common disease. PMID- 14565045 TI - [Relations between parameters which define quantity and distribution of adipose tissue]. AB - INTRODUCTION: Obesity is defined as an increase of body fat mass. Regional fat distribution, especially abdominal obesity, is a very important risk factor for cardiovascular and metabolic complications in obesity. The aim of this study was to investigate the relationship between some anthropometric parameters and body fat mass in normal weight and obese subjects. MATERIAL AND METHODS: The study group consisted of 60 obese and 60 normal-weight subjects of both sexes. All examinees underwent following anthropometric measurements: body height, body weight, waist circumference, hip circumference and sagittal abdominal diameter. We calculated values of body mass index (BMI), waist to hip ratio (WHR), waist to stature ratio (WSR) and sagittal abdominal diameter to height ratio (SAD/H). Body fat percent (FAT%) was measured using bioelectrical impedance analysis. RESULTS AND DISCUSSION: We have established that FAT% has the strongest correlation with BMI in all examined groups. Comparing with other body fat distribution parameters, FAT% had the best correlation with waist circumference and WHR. 13.33% of normal-weight women and 10% of normal-weight men presented with borderline increased body fat mass, whereas 6.67% of normal-weight women had increased values (normal-weight obesity). Normal-weight subjects had higher values of anthropometric parameters, comparing with normal-weight subjects with normal FAT%. CONCLUSION: In order to achieve precise diagnosis of obesity it is necessary, apart from certain anthropometric measurements, to establish the body fat mass, using some methods of body composition analysis. In order to identify subjects with higher risk for obesity complications, it is necessary to analyze the size of intraabdominal fat depots. We found that waist circumference, as a good predictor of specific fat distribution, also has a very good correlation with total body fat mass. PMID- 14565046 TI - [Current aspects in pharmacologic use of bile acids]. AB - EFFECTS OF BILE ACIDS AND THEIR SALTS ON ABSORPTION OF OTHER SUBSTANCES: Bile acids and their salts increase intestinal absorption of lipids and transmembrane and paracellular transfer of small and endogenous and exogenous polar molecules. It has been established that they are good promotors of insulin absorption through skin and nasal mucose, and of blood-brain barrier transfer of salycilates and quinine. EFFECTS OF BILE ACIDS AND THEIR SALTS ON ABSORPTION OF OTHER SUBSTANCES AND THEIR POTENTIAL ACTION: It has been established that combination of bile acids with amphotericin B has potential Leishmanicideal effect and combination with ciprofloxacine has improved its antibacterial activity against Pseudomonas aeruginosa in vitro. BILE ACIDS: PHARMACADYNAMIC EFFECTS: Bile acids have analgesic and hypoglycemic effect. They also have anti-HIV effect probably suppressing virus transmission from cell to cell. CONCLUSION: New studies of natural bile acids and new synthetic bile acids have revealed that they are not only adjuvants to existing active principles in pharmaceutical forms, but they can act as new therapeutic agents. However, it is necessary to study their possible mechanisms, but they are not crucial for their therapeutic application. Toxicological and pharmacological studies will determine the role of newly synthesized bile acids and their salts in current therapy. PMID- 14565047 TI - [Aging and infection]. AB - EPIDEMIOLOGY: Aging is a natural process and a part of our lives, but nowadays there is an increase in the number of persons aged 65 and over. Today infectious diseases are still responsible for one-third of all deaths in the world. The elderly population is most vulnerable to serious infections and at greatest risk for death and complications. Among geriatric population pneumonia and influenza are the fourth most common cause of death. VACCINATION: One of the goals of preventive medicine is to reduce the rate of complications and mortality from infectious diseases by increasing immunization rates. Influenza and pneumococcal vaccines are indicated for persons aged 65 and over. Despite well-recognized benefit of such vaccination, less than 50% of eligible patients receive the vaccine each year. INFECTIONS: Older persons generally have increased susceptibility to infections because of multiple risk factors and they are the most vulnerable population to nosocomial and health-care associated infections. Older persons may manifest infectious diseases atypically, with acute confusion or delirium which can lead into delay in diagnosis and therapy. It is important to know that the older present with delayed or poor response to antimicrobial therapy and high rates of adverse reactions to drugs, including antibiotics. CONCLUSION: As elderly population is rapidly growing, majority of patients with serious or life-threatening infections are old. Geriatric issues have not typically been a focus of training in infectious diseases, but we must become aware of and knowledgeable about special and unique aspects of infections in this population. PMID- 14565048 TI - [Heparin-induced type II thrombocytopenia--new views on diagnosis and therapy]. AB - HEPARIN-INDUCED THROMBOCYTOPENIA (HIT): Management of heparin-induced thrombocytopenia (HIT) and treatment options have significantly changed recently. Heparin may induce two types of thrombocytopenia. Type I, occurring earlier with a much higher rate of incidence (5-30%), is characterized by mild thrombocytopenia without significant clinical manifestations. Type II, is less frequent (0.5-2%), life threatening immune type, develops following a period of minimum 5-7 days upon introduction of heparin therapy (patients earlier treated with heparin are excluded). Type II heparin-induced thrombocytopenia with severely reduced platelet count may be clinically manifested by thrombosis in 20 50% cases within the period of 30 days. HIT is suspected in persons resistant to heparin with relatively reduced platelet count, though HIT is described in person with normal platelet counts, as well. None of available assays used for HIT detection is completely reliable. Sensitivity of a highly specific platelet aggregation assay is only 36%, sensitivity and specificity of 14C-serotonin release assays amounts to 95%, while ELISA using a heparin/platelet factor-4 target has a sensitivity of 85%. Thus, it is sometimes necessary to combine functional and antigen assays. Furthermore, new classes of antigen assays, like antibody detection tests of complexes between heparin and neutrophil-activating peptide-2 as well as those between heparin and interleukin-8, have been used. CURRENT THERAPY OPTIONS: Current therapy options exclude formerly applied low molecular-weight heparins due to the existing cross-reactivity of 80-100%. Danaparoid sodium exhibits in vitro cross-reactivity of 10-61%, clinically manifested in less than 5% of patients. Two drugs are drugs of choice in HIT type II treatment: lepirudin, especially in patients without renal failure, and argatroban, particularly in patients with renal failure. The following procedures and agents are also efficient: asmapheresis in the first four days, high-dose intravenous gammaglobulin, antiaggregans, especially ADP antagonists, aspirin, dipirydamole, dextran, prostacyclin analagoues, thrombolytic therapy as well as thromboembolectomy. Oral anticoagulants are not administered in active HIT type II, in deep vein thrombosis with high international normalized ratio (INR) and thrombin-antithrombin complexes, and low protein C levels to avoid the possibility of venous limb gangrene development. They can be administered in a stable phase, when the thrombin generation is controlled by previous administration of one of the above-mentioned alternative anticoagulants. PMID- 14565049 TI - [Specific aspects of psychoses in mentally retarded children and adolescents]. AB - Mental retardation is a heterogenous neurodevelopmental disorder characterized by arrested or incomplete psychological development. The first part of the study deals with psychological and biological factors: etiology and pathogenesis of mental retardation and comorbid psychiatric disorders. Their etiopathogenesis is similar as in other neurodevelopmental disorders and it was analyzed in the part dealing with biological specificities of persons with mental retardation. Numerous biopsycho-social factors cause increased vulnerability of the mentally retarded to development of mental disorders. Thus, prevalence of these disorders is higher in mentally retarded persons than in general population. This study also deals with specificities regarding diagnosis of psychotic disorders in mentally retarded persons as well as neurobiologic, epidemiologic, clinical and therapeutic characteristics of schizophrenic psychoses, autism and affective disorders in persons with mental retardation. Special emphasis was given to diagnostics of these disorders in mentally retarded children and adolescents, as well as to problems of differential diagnostics. Apart from other things, we have concluded that specific clinical pictures demand subspeciality approach in the frame of developmental psychiatry. PMID- 14565050 TI - Ureteral injuries in gynaecologic oncology surgery procedures. AB - This study presents ureteral injuries in gynecologic oncology surgical procedures performed at the Department of Obstetrics and Gynecology in Novi Sad, in the period from 1991 to 2001. Intraoperative ureteral injuries were recorded in 4 (1%) patients, including: partial ureteral dissection, bilateral ureteral ligation and complete ureteral dissection bellow the lower pole. In 3 (75%) cases, injuries were recognized immediately, and surgical reparation was performed. The patient with bilateral ureteral ligation presented with increased creatinine levels, anuria and development of hydronephrosis detected by ultrasonography 24 h later. Leading reasons for operative ureteral injuries include inadequate experience of surgeons, carelessness during surgery, wrong indication and wrong approach to ureter. Preparation of ureter with minimal trauma, precise localization and visualization of the complete ureteral length, as well as preservation of its vascular network, are of utmost importance in prevention of injuries. PMID- 14565051 TI - [Prognostic significance of tumor-induced angiogenesis in colorectal carcinoma]. AB - INTRODUCTION: Tumor-induced angiogenesis is a central pathogenic step in the process of tumor growth, invasion and metastasis. The aim of this study was to analyze the quantitative expression of angiogenesis in colorectal carcinoma and to determine if and how angiogenesis correlates with other clinicopathologic factors and prognosis. MATERIAL AND METHODS: This study included 40 patients who underwent curative resection of colorectal cancer at the Department of Surgery of the Senta General Hospital with complete 5 years follow-up or till death. Microvessels were identified immunohistochemically, using monoclonal CD31 antibodies. The microvessel count was assessed by means of stereology with test grid M42, as well as vascular surface density in the stromal volume at the invasive front of colorectal cancer. RESULTS: Tumor-induced angiogenesis count of colorectal carcinomas statistically significantly correlated with stage of disease and histologic tumor grade. There was no significant correlation between intratumoral microvessel density and sex and age of patients, localization and histologic tumor type. Five-year survival rate in patients with hypervascular colorectal tumors was statistically significantly lower than in patients with hypovascular tumors. Thus, microvessel density in colorectal cancer is an independent prognostic factor, but its significance is less than the importance concerning stage of disease and histologic grade of tumor. CONCLUSIONS: Intratumoral microvessel density quantification in histologic specimens of colorectal carcinoma reflects the biological malignant potential of tumors and may be a useful additional predictive marker. Assessment of intratumoral microvessel count might be used for determining the pathologic stage when adjuvant therapy is concerned. Microvessel density in tumor specimens is valuable in stratifying patients in planning appropriate adjuvant and antiangiogenic therapy after surgery. PMID- 14565052 TI - [Septic arthritis in neonates and infants]. AB - INTRODUCTION: Septic arthritis represents an intra-articular infection caused by pyogenic bacteria. During the earliest childhood it is considered to be a systemic septic condition and demands early diagnosis and prompt surgical treatment. MATERIAL AND METHODS: This is a retrospective analysis of patients with septic arthritis treated at the Department of Orthopedics of the Pediatric Surgery Clinic in Novi Sad, over a 10-year period. We are also presenting a case of a 12-day-old newborn baby, with clear radiological signs of osteoarthritis of the right knee. RESULTS: A retrospective study included the period 1991-2000, and showed that 15 patients, aged 10 days--12 months were treated for osteoarthritis. The most common localization was the hip, in 60% of cases. In 11 patients the causative agent was Staphylococcus aureus, while in the 4 remaining patients the bacteriologic finding was negative. One patient died of generalized sepsis. DISCUSSION: In neonates and infants septic arthritis is characterized by atypical clinical picture, often causing delayed diagnosis. In the initial phases of the disease ultrasonographic findings were of greater use compared to radiological imaging, due to relatively late appearance of radiological signs of disease. CONCLUSIONS: Due to possible development of serious and irreversible damage, even lethal outcome, septic arthritis requires early diagnosis, prompt administration of antibiotics and early surgical treatment. It is a quite unique area in Pediatric Orthopedics where missed or delayed diagnosis may have serious consequences. PMID- 14565053 TI - [Dyslipidemia and subclinical hypothyroidism]. AB - INTRODUCTION: Subclinical hypothyroidism is defined as an increased serum TSH and normal serum FT4 concentration. In subclinical hypothyroidism, thyroid peroxidase and thyroglobulin antibodies are frequently present. Subclinical hypothyroidism may have endogenous or exogenous causes. The prevalence of subclinical hypothyroidism is rather high. The number of patients progressing to overt hypothyroidism may be higher. These patients may be asymptomatic, or have only mild symptoms or a single symptom. MATERIAL AND METHODS: We investigated 35 patients with subclinical hypothyroidism in order to establish the type and degree of dyslipidemia and effects of therapy with L-thyroxine (50 micrograms/d) during three months. RESULTS: Serum cholesterol, LDL-cholesterol and apo B were increased. A significant reduction of serum cholesterol, LDL-cholesterol and apo B concentrations was established during thyroid hormone replacement. DISCUSSION AND CONCLUSION: Only a few studies reported higher LDL and lower HDL-cholesterol values in subclinical hypothyroidism. Much interest was thus aroused to evaluate whether or not subclinical hypothyroidism is associated with hypercholesterolemia. Only patients with serum thyrotropin (TSH) concentration above 10 mU/L had a significant reduction of serum cholesterol concentration during thyroid hormone replacement. Most patients with subclinical hypothyroidism should be treated with thyroxine to prevent progression to overt hypothyroidism. Thyroid hormone replacement therapy may slow the progression of coronary heart disease, because of its beneficial effects on lipids. These findings and especially high rate of progression towards overt hypothyroidism suggest early thyroxine treatment. PMID- 14565054 TI - [Major characteristics of immunoglobulin A nephropathy]. AB - INTRODUCTION: Immunoglobulin A nephropathy (IgAN) is one of the most common forms of primary glomerulonephritis in many countries. Most clinical features of IgAN point to a renal problem, such as recurrent macroscopic hematuria or asymptomatic microscopic hematuria and proteinuria. Pathologic features of IgAN present with different types and different degrees of glomerular, tubulointerstitial and vascular lesions. The aim of this study was detailed analysis of clinical and laboratory findings, as well as findings of immunofluorescence and light microscopy. We also investigated associations between these factors. MATERIAL AND METHODS: We investigated 60 patients who underwent renal biopsy. The study was partly retrospective and partly prospective. RESULTS: The average age of patients was 34.19 years. Male female ratio was 2.33:1. IgAN was most frequently asymptomatic (83.33%) as microhematuria and proteinuria, while gross hematuria was found in 16.667%. Renal biopsy material was analyzed by light microscopy revealing changes in all glomerular structures. Immunofluorescence microscopy demonstrated dominant IgA deposits. This study established association of glomerulosclerosis with clinical features of disease. DISCUSSION AND CONCLUSIONS: IgAN frequently develops in the 4th decade of life, mostly in males and presents as asymptomatic (83.33%). Pathohistological changes include all glomerular structures. There is no specific serological test for IgAN, but pathological changes affect clinical features of the disease, as proteinuria and increase of creatinine concentration. PMID- 14565055 TI - [Reconstruction of the esophagus with a jejunal segment--case report]. AB - INTRODUCTION: Portions of gastrointestinal tract may be used as microvascular transfers for reconstruction of hypopharynx and esophagus. Colon, jejunum and portions of stomach have also been successfully used. Intestinal transfer by revascularization was reported by Seidenberg in 1959, long before the development of other free tissue transfers. MATERIAL AND METHODS: When an appropriate piece of jejunum is being selected for transfer, the important considerations are its shape and its vascular supply. Because of multiple curves of jejunum, it is difficult to obtain straight lengths longer than 12-15 cm without separating it from its mesentery at both ends. DISCUSSION: Mucous production is very important in the first postoperative days. When edema decreases and patients can swallow fluids well, oral intake can gradually be increased to full fluids and eventual regular diet. It is important to monitor jejunum following its transfer. Anastomosis thrombosis and unrecognized gangrene of jejunum may cause serious neck infections. CONCLUSION: Portions of gastrointestinal tract may be used as microvascular transfers for reconstruction of hypopharynx and esophagus. Intestinal transfer by revascularization was reported by Seidenberg in 1959. It is important to monitor jejunum following its transfer. PMID- 14565056 TI - [Umbilical hernia in children]. AB - INTRODUCTION: Umbilical hernia is a common condition in the pediatric population. EMBRYOLOGY: Umbilical hernia is a consequence of incomplete closure or weakness at the umbilical ring, where protrusion of intraabdominal contents may occur. ANATOMY: Fascia posterior to the canal is thinner, creating an area of weakness. Congenital or direct hernia occurs in this area, while herniation in the umbilical canal leads to indirect or acquired hernia. INCIDENCE: The incidence of umbilical hernia is 1.9% to 18.5% in white population. CLINICAL MANIFESTATIONS: The great majority of pediatric umbilical hernias are asymptomatic. Incarceration and strangulation are uncommon. Rupture of umbilical hernia with resultant evisceration is extremely rare. Umbilical hernia may also be the source of intermittent umbilical or abdominal pain. TREATMENT: Treatment options for umbilical hernias range from simple observation to surgical repair. The great majority close spontaneously and observation with periodic follow-up is appropriate in most cases. There are no available data to suggest that strapping improves or accelerates closure. Operation would be recommended for defects greater than 1 cm, by the age 3 to 4. Persistence or enlargement of fascial defect during the period of observation are reasons to consider repair, whatever the age. COMPLICATIONS: Complications of operative repair of umbilical hernias include those related to anesthesia and local wound infections. CONCLUSION: Umbilical hernia is a common condition among infants and children. In the great majority of cases, the natural history is one of eventual closure without treatment. If spontaneous closure does not occur until the age of 3-4 years, operative correction is recommended. PMID- 14565057 TI - [History of scurvy therapy and the injustice done to Dr. Maksim Nikolic Miskovicev of Sremski Karlovci in the 19th century]. AB - This paper reviews historical aspects regarding medical knowledge on scurvy, vitamin C deficiency, and its etiological factors. This disease used to affect naval-crews on long lasting sea voyages, soldiers during times of war and the poor. Although efficient therapy of scurvy (fresh vegetables, lemon juice) was known in the mid-18th century, its etiology was not known. It was believed to be caused by the cold, moist winds, unhealthy evaporations, malnutrition and it was called "alkaline disease". It was established that acid substances like lemon juice, had beneficial effects on the disease. However, it was soon generally accepted that it was caused by deficient diet. In 1830, Dr. John Elliotson, Professor of Medicine at London University supported this opinion. In 1928, Albert Szent Gyordyi, professor of Chemistry and Biochemistry at Szeged and Budapest Universities, isolated vitamin C from green pepper, vegetable proved to be rich with vitamin C. Due to this discovery, and some discoveries regarding biological processes, he was awarded the Nobel Prize for medicine and physiology in 1937. Back in 1864, a military doctor in Belgrade, Dr. Maksim Nikolic Miskovicev from Sremski Karlovci, wrote to Ministry of Defence of the Serbian Principality. He informed the authorities about a successful, quick and cheap cure for scurvy-pepper juice. A medical board headed by Dr. Vladan Dordevic was formed to evaluate his discovery. Dr. Nikolic-Miskovicev was underestimated and laughed at, and his discovery was completely forgotten. The aim of this paper was to correct injustice done to Dr. Maksim Nikolic-Miskovicev. PMID- 14565058 TI - [Epidemiology of decision on life-sustaining treatment in the general internal medicine division]. AB - In the state of Israel there are no clear guidelines of decision-making for terminally ill patients. However, physicians do make decisions concerning life sustaining treatment. The rate of these decisions in medical wards in Israel is not known. The purpose of the study was to investigate prospectively, in a medical division, the rate of foregoing life-sustaining treatment. In addition, we wanted to find out whether or not senior physicians and families were involved in the decision-making process. The relationship of the decisions to the patients demographic and clinical characteristics was evaluated. Of 1039 patients hospitalized in 3/01, 12/01 and 1/02, the decision to forego life-sustaining treatment was taken in 8.8% of the cases. The highest rate was observed in the acute geriatric ward (23%) and the lowest in the Department of Cardiology (< 1%). Senior physicians were involved in 74% of the decision-making and the family was involved in only 29%. Age, marital status, dementia, functional state and the severity of sickness significantly affected the decision-making. However, on the basis of multi-regression analysis, only functional impairment prior to the hospitalization significantly affected the decision-making by the physician. The findings of this study support the essential need for accelerating legislation concerning decision-making for patients at the end of life. The adoption of the recommendations of the public committee on this subject, recently presented to the Minister of Health, provide legal validity to advance medical directives and may assist in ethical issues for end of life decision-making. PMID- 14565059 TI - [Stapled hemorrhoidectomy--early experience in 30 patients]. AB - INTRODUCTION: We report the early results of 30 patients treated by stapled hemorrhoidectomy. PATIENTS AND METHODS: Thirty patients with symptomatic grade 2 4 hemorrhoids were treated by stapled hemorrhoidectomy. The procedure was performed with a 33 mm diameter automatic hemorrhoidal circular stapler. The patients were prospectively evaluated for immediate and functional recovery, postoperative pain and subjective success of the treatment. RESULTS: The median age of the patients was 46.2 years, and the median duration of the symptoms was 27 months. The main symptom was bleeding (in 96% of the patients). The average operative time was 22 minutes. The operation was performed with spinal (63.3%) or general (36.7%) anesthesia. There was no mortality, urinary retention, incontinence, fecal urgency or persistent pain. One patient, who was under anticoagulant treatment, had postoperative bleeding, which required transfusion of 3 units of blood, and another patient was operated on because of perforation of the sigmoid colon. Most patients (twenty eight) had complete functional recovery and returned to their usual daily activities within 10.4 days. Postoperative pain and subjective success were evaluated by a 1 to 10 scale. The average pain score decreased from 5.8 on the first postoperative day to 2.4 on the 7th postoperative day. The average satisfactory score was 9.2. CONCLUSION: Stapled hemorrhoidectomy is an alternative to conventional surgical hemorrhoidectomy. The procedure seems to be associated with less postoperative pain and early recovery with a high satisfaction rate among patients. PMID- 14565060 TI - [Reduction of intussusception by air enema in children--experience over a 13-year period]. AB - PURPOSE: Intussusception is a relatively common pediatric emergency. The accepted treatment is closed reduction using barium or air enema and surgery when it has failed. We present our 13-year experience using air enema for treatment of intussusception, emphasizing the reduction with repeated delayed air enema. METHODS: Between February 1990 and December 2002, air enema procedures were performed in pediatric patients in cases with suspected intussusception. Prior to the procedure, all patients were sedated with meperidine hydrochloride (0.5 mg/kg) or morphine (0.1 mg/kg). Before 1993, surgery was performed after the first ineffective attempt of air enema reduction. Since 1993, in the case of failure of the first attempt, two additional trials of air reduction were performed at an interval of 45-60 minutes prior to surgery. RESULTS: A total of 225 air enema procedures were performed in 201 children (129 males, aged 10.4 +/- 9.1 months, range 8-63 months) with suspicion of intussusception. The diagnosis was confirmed in 148 (66%) out of the 225 procedures performed. A successful reduction of intussusception (by one attempt or more) was achieved in 125 (86%) out of the 148 procedures with proved intussusception. Twenty-one (14%) patients were operated on after the failure of closed reduction. Spontaneous reduction was demonstrated under fluoroscopy in two (0.8%) studies. No complications of the air enema were noted. During 1990-1993, a single reduction attempt of the intussusception by air enema was performed. Successful reduction during this period was achieved in 19 (70%) out of 27 patients and 8 (30%) children were operated on. During the period 1993-2002, we used repeated delayed attempts to obtain reduction of intussusception among 174 patients. The success rate was 89% (108 out of 121 patients) and only 13 (11%) children were operated on. CONCLUSION: Air enema is an efficient and safe diagnostic and therapeutic tool for intussusception in the pediatric population. PMID- 14565061 TI - [Sexual dysfunction--what is the primary physicians role?]. AB - BACKGROUND: Sexual dysfunction is characterized by disturbances in sexual desire and/or psycho-physiological changes associated with the sexual response cycle in men and women. Studies suggest that it is prevalent in 10-52% of women and 25-63% of men, and could have a tremendous impact on the individual, interpersonal relations, and on the family quality of life. OBJECTIVE: To assess attitudes, knowledge and sexual dysfunction management by Israeli primary care physicians. METHODS: Two hundred and ninety-two physicians from approximately 800 participants who attended a Family Medicine and General Practice conference took part in the study. A self-reply questionnaire was distributed among the participants and 179 completed the full questionnaire (61%). RESULTS: Seventy nine percent of the physicians thought the primary care physician should be the address for treating most sexual dysfunction problems. Physicians indicated that only 12% of their patients treated for sexual dysfunction were female. Significant differences were found between male and female physicians, board certified family physicians, general practitioners and residents in family medicine regarding sexual dysfunction treatment. Board certified family physicians treated more patients with sexual dysfunctions than general practitioners (GP) and residents in family medicine [53% vs. 23% vs. 22% respectively, (p < 0.05)]. The main barriers for treating sexual dysfunctions noted by physicians were lack of time (62%) and knowledge (47%). CONCLUSIONS: Family physicians consider themselves to be the main address for the patient for sexual dysfunction problems. Physicians believe they need more training to overcome lack of knowledge barriers and they need to organize their time to adequately address their patients sexual dysfunction problems. PMID- 14565062 TI - [Medical termination of pregnancy with mifepristone--initial experience at the Sheba Medical Center]. AB - OBJECTIVE: To assess the early implementation of medical termination of pregnancy as an alternative treatment option for women in early pregnancy who wish to avoid a surgical procedure. METHODS: The use of mifepristone (RU-486) for medical termination was recently approved in Israel. We performed a prospective follow-up study of the 189 women who underwent medical termination of pregnancy with mifepristone and misoprostol in our center between January 2000 and April 2001. RESULTS: Complete outcome data was obtained for 176 women. Within this group 152 women (86.4%) did not need any surgical procedure to complete the abortion. Uterine curettage was performed in 17 (9.6%) and operative hysteroscopy in 7 (4.0%) of the women because of incomplete abortion or suspected residua of pregnancy. CONCLUSION: Medical abortion offers an efficient and safe treatment option to women who wish to avoid surgical evacuation. PMID- 14565063 TI - [Chest wall deformity following rib cartilage harvesting for auricular reconstruction]. AB - Chest wall deformity is one of the potential complications of rib cartilage harvesting for auricular reconstruction. An eight year old boy underwent two stage microtia reconstruction using the 6th-9th costal cartilage for the 3-D cartilage framework. One year later an anterior chest wall deformity was noticed with bulging of the 5th rib, asymmetry of the lower rib cage line and deviation of the lower part of the sternum. The anterior chest wall is made of ribs, sternum and respiratory muscles forming a dynamic structure. By using these pieces of rib cartilage an imbalance is created in which the respiratory muscles pull the edges of the remaining ribs, thereby creating the deformity. It is especially critical during the growth phase. The 3-D CT scan demonstrates the chest deformity. We present a literature review and recommendations on how to reduce the complications when using rib cartilage for auricular reconstruction are presented. PMID- 14565065 TI - [The importance of the Cochrane Collaboration]. PMID- 14565064 TI - [End-of-life decision-making process]. AB - Since time immemorial the attitude toward the dying patient has been one of the most difficult issues in medical ethics. The diversity of philosophical, religious, social and legal approaches does not enable one to reach a universal consensus to solve the many problems involved in end-of-life decisions. Within the health care system in Israel there is currently no consensual practice concerning the dying patient. Moreover, there is no published information on the actual decision-making processes within hospitals, hospices and geriatric facilities in Israel concerning the dying patient. A group of investigators in Shaare Zedek Medical Center in Jerusalem recently performed a prospective study to explore the decision-making process concerning DNR orders within this hospital. The results of this study demonstrated that the terminally-ill patients never take part in the decision-making process, they are never consulted about their wishes, and there is no effort to discover their previous wishes concerning their treatment at the terminal stages. Moreover, in many instances even the family was not consulted and did not take part in the decision-making process. In a significant minority, the final decision of a DNR order was undertaken by a single physician. This approach represents an extreme form of unethical paternalism, and it requires an urgent societal intervention to establish an ethically sound decision-making process. Recently, a national committee ("Steinberg Committee") formulated a widely agreed upon legislative proposal organizing all the fundamental and practical issues related to the dying patient. This proposal is based upon a balance between opposing values such as autonomy, life, quality-of-life, beneficence, non-maleficence and "slippery-slope" concerns. It relates to various treatment modalities, such as resuscitation, ventilation, dialysis, medication and sustenance. It establishes a clear position on euthanasia, physician-assisted suicide, withdrawing treatment and withholding treatment. It establishes a hierarchy of decision-making agents, and it validates advanced medical directives. It also promotes a legally-binding requirement of modern palliative care. PMID- 14565066 TI - [Treatment of hypertension with device-guided breathing exercise]. AB - Hypertension has been documented as a major risk factor for cardiovascular morbidity and mortality and lowering blood pressure (BP) can reduce this risk. Life style modifications such--as physical exercise, salt restriction and weight reduction can lower BP. Recently, a number of studies showed that device-guided breathing exercise, which lowers the respiration rate and modifies respiratory patterns, can also lower BP. Deep and slow breathing increase baroreflex sensitivity and heart rate variability, improve small vessels blood flow, and decrease peripheral resistance, thereby leading to BP reduction. Several studies including 268 hypertensive patients showed that 15 minutes of daily breathing exercise lowered BP within 8 weeks by 12.1/6.1 mmHg as compared to 7.6q3.4 mmHg in the control group. Most patients complied with the treatment. These results suggest that device guided breathing exercise can be used to lower BP. PMID- 14565067 TI - [Cervical carcinoma associated with pregnancy]. AB - Cervical carcinoma is one of the most common malignancies diagnosed during pregnancy since about 30% of diagnosed women are in their fertile years. Diagnosis of cervical cancer during pregnancy superimposes ethical dilemmas on the clinical considerations. Both maternal and fetal outcome should be considered when managing such a case. Is it necessary to discontinue the pregnancy? What are the implications of delayed treatment? How does the treatment affect the fetus and newborn? This article reviews the practical aspects of these issues and suggests management proposals. PMID- 14565068 TI - [Spinal cord injury due to penetrating missiles]. AB - Gunshot wound of the spine is a major cause of spinal cord injury among US civilian population, members of the military armed conflict personnel, or civilians injured in terrorists attacks. The bullet fragments cause damage to the spinal cord even without penetrating the spinal canal. Concussive effects, heat, fractures or vascular injury may cause the neurological damage. Unfortunately, bullet or shrapnel removal or laminectomy do not change the prognosis. In this article we review the historical background, the Israeli experience, ballistic forensic considerations, complications, treatment and prognosis. PMID- 14565069 TI - [Updates in osteogenesis imperfecta]. AB - Osteogenesis Imperfecta, a heterogenous group of inherited disorders, is characterized by both bone fragility and low bone mass. Since the Sillence classification of the disease, significant progress has been made in understanding the molecular, histological and genetical pathogenesis of the disease, and it led to expanded nosology. The multidisciplinary approach including rehabilitation, medications and surgery, may significantly improve the quality of life and prolong life expectancy. PMID- 14565070 TI - [Growth and development aspects of international adoption]. AB - With the increase in the number of children being adopted by Israeli families from abroad, the medical system has to face heretofore unfamiliar medical and developmental challenges. Many of the biological mothers of these children were substance abusers and suffered from a variety of illnesses and nutritional deficiencies during their pregnancy. Their infants and young children were placed in orphanages at a very young age. The ensuing emotional and environmental deprivation in many cases impacted negatively on their neurological, physical and emotional development. Whereas our previous survey dealt with the purely medical aspects of international adoption, the current review deals with the growth and developmental issues involved in raising these children. PMID- 14565071 TI - [Iron deficiency and anemia in female athletes--causes and risks]. AB - Iron deficiency is probably the most common nutrient deficiency in the western world. Low levels of iron in the body are caused by several mechanisms, and become symptomatic with the onset of iron deficiency anemia. Athletes are a special group with additional reasons for iron or blood loss, such as plasma expansion, increase perspiration, 'foot strike hemolysis, and occasionally- malnutrition. Female athletes have yet another source of blood loss- menstruation. However, the most common cause for low hemoglobin levels in an athlete is dilutional pseudoanemia, which is caused by exercise-induced fluid retention. Athletes are more sensitive to the effects of anemia and iron deficiency, as exercise performance depends on maximal oxygen carrying capacity to the active muscle, and efficient oxygen utilization. Iron deficiency without anemia can also reduce athletic performance. Diagnosis is ultimately made by a blood count and red blood cell parameters, with ferritin serving as an index of body iron stores. Treatment requires iron supplements, as it is nearly impossible to refill the iron stores through diet alone. PMID- 14565072 TI - [Physical activity and training at high altitude]. AB - In recent years there has been a growing interest in the effects of high altitudes on the human body. High altitudes are being frequented by more and more people for sport and leisure pursuits, and are increasingly being used both as a training environment and for investigating the healthy body in hypoxic conditions. During the ascent from sea level, atmospheric pressure and partial oxygen pressure decrease, humidity and temperature decrease, and radiation is elevated. The altitudes at which physiological changes and clinical symptoms occur are not constant, but variations may usually appear above 2300-2800 meters. There is a wide variability of reactions to low oxygen in the air inhaled. The physiological parameters during both rest and physical activity at high altitudes are different from those at sea level, and the differences are reflected in concomitant changes in attitude and behavior. Research into altitude as a training environment that could improve athletic endurance and performance has grown in the last decade, with the development of a number of new training methods, such as "Living High--Training Low" and "Training High--Living Low". These have contributed to an improvement of performance in a significant number of athletes. This article will demonstrate the impact and possible dangers of physical activity at high altitude, and will present the current knowledge and methods of altitude training. This article reviews the influence of low air and oxygen pressure on athletic performance. Recommendations are presented regarding physical activity at high altitude and nutritional support. PMID- 14565073 TI - [The 60th conference held by the American Academy of Allergy Asthma and Immunology (AAAA), on March 7-12, 2003]. PMID- 14565074 TI - [Lipid peroxidation in adult and aged rat heart under immobilization stress]. AB - Lipid peroxidation (LPO) in the heart of adult (10-12 month) and old (22-25 month) Wistar male rats was investigated during immobilized stress. Immobilization of rats was accompanied by increasing the concentration of all studied LPO intermediates in the myocardium of both age groups. Accumulation of LPO products in old rat hearts was considerably higher than in adult ones. Dimethyl sulfoxide injection limited stress-induced stimulation of LPO. There were age related variations in efficiency of this antioxidant effect on the heart during immobilized stress. PMID- 14565075 TI - [Effect of a low concentration of hydrogen peroxide on metabolism of blood cells]. AB - Hydrogen peroxide is known to posses a wide range of physiological effects towards functional activity of cells. We have investigated the influence of H2O2 on the activity of glycolys in the native blood cells. Adding of hydrogen peroxide up to final concentration 50 microM led to decrease of activity some glycolytic enzymes. H2O2 inhibited consumption of glucose by blood cells. Despite the fact that H2O2 is well known as prooxidant, we observed decrease of the level of thiobarbituric acid reactive species in the blood cells after their incubation in the presence of hydrogen peroxide for 2 hours. Based on these data we supposed that effects of hydrogene peroxide occur via signal transduction pathways. PMID- 14565077 TI - [Structure-functional status of the erythrocyte membrane in acute poisoning by metaphos and administration of perftoran]. AB - The effect of perftoran administration on structure-functional state of erythrocyte membrane was investigated under conditions of metaphos intoxication. Metaphos administration at the dose of LD50 caused pronounced membrane-toxic effect. Perphtoran administration to animals reduced manifestation of the membrane damage. PMID- 14565076 TI - [Some factors of collagen aging factors in vivo and in vitro]. AB - The literature data on different factors of collagen aging, the main connective tissue protein, have been analyzed. Mechanisms of reactions that are involved in both old aging and non-enzymatic course of protein aging are discussed. Factory influencing, non-enzymatic protein glycation during-related pathology and some ways for its inactivation by different substances are reviewed. PMID- 14565078 TI - [Expression of the Mycobacterium tuberculosis cytochrome CYP51 gene in Escherichia coli]. AB - Mycobacterium tuberculosis sterol 14 alpha-demethylase CYP51 is currently considered a promising target for a new generation of antitubercular drugs. Similarly to eucaryotic sterol 14 alpha-demethylases, candidate selective inhibitors of M. tuberculosis CYP51 belong to azoles, the N-substituted derivatives of imidazole and triazole. We have constructed a highly effective system for heterologous expression of MT CYP51 in E. coli. The recombinant protein was purified by metalloaffinity chromatography for primary in vitro screening of novel antitubercular azole compounds for their high affinity to the molecular target. Yield of the purified recombinant MT CYP51 was about 1.5 micromoles of the purified native protein per 1 l of E. coli cell culture. The recombinant protein interacted with ketoconazole with a Kd of 7.7 microM. MT CYP51 produced by our system for heterologous expression in E. coli may be used for initial testing of novel antimycobacterial azole drugs. PMID- 14565079 TI - [Principles of enzymatic analysis of lipids]. AB - The review described principles of modern methods of enzymatic analysis of lipids. Many of these methods of determination of cholesterol, cholesterol ethers, triglycerides, free fatty acids, various types of phospholipids and oxisterols may be employed or are employed in clinical diagnostic laboratories. PMID- 14565080 TI - [Antioxidant properties of a leaf extract from Aronia (Aronia melanocarba) containing proanthocyanidins]. AB - Antioxidant capacity of a procyanidin-containing extract from Aronia melanocarpa leaves has been studied in vitro and in vivo. Using the chemiluminescence technique, the effective content of antioxidants and its reactivity towards peroxyl radicals have been measured in a model reaction of initiated oxidation of hydrocarbon for the whole extract and two of its chromatographic fractions separated by HPLC. The results indicate that the extract contains a combination of antioxidants with different radical scavenging activities. The value of the rate constant of the reaction with peroxyl radicals (constant k7) for the extract strong inhibitors is of the same order of that for alpha-tocopherol and its synthetic analog chroman C. A decrease in antiradical activity directly related to the high concentrations of this extract in reactive mixture has been observed. This extract significantly reduced in dose-dependent fashion the CCl4-induced hepatic lipid peroxidation. The extract also inhibited flaxseed oil peroxidation. The present study indicates significant antioxidant capacity of the extract from Aronia melanocarpa leaves. PMID- 14565081 TI - [Paramagnetic spectrum and biological activity of humic series peloid preparations]. AB - Peloid preparations may influence activity of some oxidoreductases (lactate dehydrogenase, malate dehydrogenase). Their effects depend on physico-chemical properties of the compounds tested. PMID- 14565082 TI - [Structure-functional change in streptokinase exposed to ultrasound]. AB - To optimize conditions of acousto-enzymatic thrombolysis the influence of low frequency ultrasound on the isolated preparation of streptokinase was investigated. The ultrasound treatment with intensity 26 W/cm2 at 37 degrees C within 5-10 minutes was not accompanied by changes of structure-functional properties of the streptokinase molecule. Increase of ultrasound-processing time (10-60 minutes) resulted in non-covalent hydrophobic aggregation of some part of the protein. In contrast to native protein ultrasound modified streptokinase is readily degraded by plasmin with formation of polypeptide fragments with molecular weights ranged from 43 up to 14 kD. The processes of aggregation and increased proteolytic degradation resulted in lower efficiency of plasmin autoactivation under the action of sounded streptokinase. PMID- 14565083 TI - [Inhibition of oxidation of human blood low density lipoproteins by carotenoids from paprika]. AB - beta-Carotene (C1), acyl derivatives of capsanthin (C2), and acyl derivatives of capsorubin (C3) were isolated from red paprika (Capsicum annuum) oleoresin. Incorporation of carotenoids C1, C2, and C3 into human plasma LDLs changed the LDL flotation coefficient distribution for LDL fraction 1.019 < d < 1.050 g/ml. The comparative studies of Cu(2+)-catalyzed oxidation for LDL, LDL-C1, LDL-C2, and LDL-C3 revealed that carotenoids C1, C2, and C3 efficiently suppressed the oxidation of LDL, inhibited the formation of conjugated dienes from polyunsaturated fatty acid residues, lowered the content of small dense LDL subfraction, and inhibited the transformation of cholesterol to auto oxidized products (in particular to 5,6-epoxycholesterol, 7-ketocholesterol and 7 beta hydroxycholesterol). This suggests that the main carotenoids may effectively inhibit LDL oxidation in vitro with probable lowering the "atherogenic" LDL subfraction production in vivo. PMID- 14565084 TI - [Comparative evaluation of the antioxidant and antiatherogenic effect of natural and selenoorganic substances]. AB - The antioxidant and atherogenic effects of naturally occurring compounds (soya isoflavones) and of the synthetic organic compounds of selenium (selenopyran and dimethyl-pyrosalyl-selenide) on the primary cell culture of human aortic subendothelial intima and on macrophage cell culture were investigated. Our results suggest that soya isoflavones exhibit mainly antiatherogenic effect and organic compounds of selenium exhibit mainly antioxidant effect. PMID- 14565085 TI - [Level of interferon-gamma, tumor necrosis factor alpha, and antibodies to them in blood serum from Parkinson disease patients]. AB - Serum levels of interferon-gamma (IF gamma), tumor necrosis factor alpha (TNF alpha) and autoantibodies (a-AT) to these cytokines were investigated in patients with Parkinson's disease (PD). The increased levels of TNF alpha (50%) and IF gamma (35%) were found in PD patients. There was close correlation between the serum level of TNF alpha and the manifestation of neurological symptoms (r = 0.434; p < 0.05), and between levels of IF gamma and the duration of this disease (r = 0.4511, p < 0.05) and patients age as well (r = 0.4358; p < 0.05). There was increased level of a-AT to TNF alpha in PD patients versus healthy controls (130.3 +/- 11.92 and 105.3 +/- 4.62, respectively, p < 0.05). The combined increase of levels of a-AT to TNF alpha and IF gamma (r = 0.91, p < 0.01) close reverse correlation between duration of PD and levels of a-AT to TNF alpha and IF gamma (r = 0.4644 and r = 0.606, respectively, p < 0.01) were also recognised. The data obtained suggest the involvement of TNF alpha and IF gamma into the pathological process during PD, which requires further investigation in this direction. PMID- 14565086 TI - [ATP-generating creatine kinase: a new compound isolated from Viperidae venom]. AB - This is a first report on the presence of creatine kinase, ATP-generating enzyme, in the venom of Vipera xanthia representing the Viperidae family of poisoning snakes. Kinetic and catalytic properties of 40 kD active monomer has been described with a following discussion on the issue of potential of the data listed for further research in either toxic coagulopathy molecular mode or enzyme application dealing with "thrombin-thrombin receptor" interaction studies. An enzyme purification procedure includes hydrophobic (phenyl-Sepharose), anion exchange (MonoQ) and affinity (ADP-Sepharose) chromatographic steps. PMID- 14565087 TI - [Spanish society of pediatric surgery in internet: the beginning of another stage]. PMID- 14565088 TI - [Predictive value of prenatal MRI in the diagnosis of thoracic congenital malformations]. AB - INTRODUCTION: Magnetic imaging (MI) has an increasing value in the prenatal diagnosis of thoracic malformations. MATERIAL AND METHOUS: We compare in this work the prenatal diagnoses with the prenatal sonographic diagnoses and postnatal imaging, surgical or postmortem findings. RESULTS: Prenatal sonography diagnosed 5 diaphragmatic hernias (CDH) and 3 cystic adenomatoid malformations (CAM). MI confirmed left side CDH in 4 cases, in two of them showing also herniation of the left hepatic lobe and the spleen. In the 5th case, MI suggested diaphragmatic eventration with partial occupation of the right hemithorax by the liver. Two of three CAM appeared to have lung sequestration at MI. At birth, four CDH and one diaphragmatic eventration were confirmed by simple x-ray, and by surgery in all but one, a CDH case who went into ECMO and died without surgery. Pulmonary sequestration was postnatally confirmed by CT scan and arteriography. Treatment was coil embolization of the systemic artery. CAM was confirmed postnatally through plain chest film and CT scan. Surgical resection of the lesion was performed and the pathology exam demonstrated the presumed lesion. CONCLUSIONS: When prenatal sonography suggest a fetal thoracic malformation, MI is the way to accurate diagnosis, follow-up, prognostic evaluation and therapeutic strategy. PMID- 14565089 TI - [Early esophageal replacement in patients with esophageal atresia]. AB - INTRODUCTION: When primary anastomosis is not feasible in esophageal atresia esophageal replacement is one of the possible options. We report our experience with this approach in patients with long-gap esophageal atresia. MATERIALS AND METHODS: From 1991 to 2002 we treated 50 children with esophageal atresia. Nine required esophageal replacement because of long-gaps. Six were boys and three girls. Six had isolated atresia, and three had regular atresia with TEF. Associated malformations in 8 patients were: cardiovascular (2), anorectal (2), urogenital (5), intestinal duplication (1), and costovertebral (1). The weight at birth ranged from 1.2 to 3.95 kg (median 2.3). Patients with type I atresia had immediate gastrostomy accompanied by esophagostomy in one. Esophageal anastomosis was impossible or failed in 3 patients with type III EA, who had fistula ligation and esophagostomy. Esophageal replacement was performed at a median age of 4.95 months (range 2.3 to 18), with a median weight of 5.33 (range 2.89 to 11.5 kg.). We used gastric pull-up in 2 cases and colonic transposition in 7 with isoperistaltic left transverse colon in retromediastinal-transhiatal (6) or restrosternal (1) position. Two patients had extramucosal piloromyotomy and 4 had piloroplasty. RESULTS: All patients survive and have functional grafts. Postoperative complications were: pneumonia (1), wound dehiscence with evisceration (2) and salivary fistula (2) which closed spontaneously. In the long range, one patient was operated three times for hernia through the enlarged hiatus, one had one episode of intestinal subobstruction successfully managed with nasogastric aspiration and another one had dumping syndrome for several months. After a median follow-up of 3.15 years (range 1.6 to 6.9) all patients eat normal diets per os. CONCLUSIONS: Esophageal replacement for the treatment of infants with long-gap EA has been in our hands as good as any other option with 100% survival and good functional results. The operation can be safely performed in the first months provided that associated malformations are under control. However, like other options, this approach is not devoid of complications. PMID- 14565090 TI - [Fiberoptic bronchoscopy, 3-D reconstruction of the airway and virtual bronchoscopy in patients with airway malformations. Preliminary report]. AB - Bronchoscopy is the diagnostic gold standard in patients with airway malformations. Helical CT scan has produced studies such as virtual bronchoscopy or 3-D reconstruction of the airway. The purpose of this study is to analyze the correlation between fiberoptic bronchoscopy, virtual bronchoscopy and 3-D reconstruction of the airway in patients with airway malformations. METHODS: From January 2001 to March 2002 we evaluated the airway of 17 patients with airway malformations by means of a diagnostic protocol consisting on fiberoptic bronchoscopy, and Helical CT scan with 3-D reconstruction of the airway and virtual bronchoscopy. The radiologist had no access to bronchoscopic information. Age at study, associated cardiovascular anomalies, indications, localization, degree of diagnostic concordance, etiology and influence in treatment were analyzed. RESULTS: 20 studies were done to 17 patients whose mean age was 1.64 +/ 0.48 years (7 days-7 years). Twelve patients had associated anomalies of the aorta, pulmonary arteries or supraortic vessels. Excellent concordance was obtained in 13 cases (65%), good in 6 (30%) and poor in one (5%). 3-D reconstruction of the airway and adjacent vascular structures provided additional information in 14 cases (70%): in 3 exact length of the tracheal lesion and in 11 defined the cause of the compression. In all the cases, absolute concordance in localization was obtained. CT scan information modified treatment in 6 patients (35%). In 7 patients with associated vascular anomalies, no further image studies were done, confirming the anatomy concordance during surgery. CONCLUSIONS: 3-D reconstruction of the airway and virtual bronchoscopy are excellent diagnostic tools in patients with airway malformations, and contribute to define the etiology, length and diameter of the lesion. Excellent diagnostic correlation was obtained between analogic and virtual bronchoscopies, although further multicentric studies should be conducted. PMID- 14565091 TI - [Indications of transduodenal papillotomy for choledochal cyst]. AB - The Choledochal Cyst is a dilatation of the biliary tract whose etiology is still under debate. OBJECTIVE: This paper analyzes the cases of our group an contributes on the indications of transduodenal papilotomy in the surgical treatment of this process. MATERIAL AND METHODS: Seven cases diagnosed as choledochal cysts are presented with mean age between 3 and 10 years (5 females and 2 males). Six cases were presented with abdominal pain, four with jaundice, three presented with biliary stones and one cases was a casual discovery during an US examination. In all cases US and magnetic resonance cholangiopancreatography (MRCP) were performed as preoperative work-up. In one case an endoscopic retrograde cholangiopancreatography (ERCP) was made. In all cases intraoperative cholangiography was carried out. The surgical treatment was quistectomy and hepatico yeyunostomy Roux-Y in four cases; in one of them a transduodenal papilotomy was added. Cholecystectomy and transduodenal papilotomy was made in three cases, two of them presented choledocholithiasys. RESULTS: All patients are symptoms free after a mean follow up period of 2 years and 3 months (range: 1 year and 6 months to 4 years and 10 months). In one case persists dilated choledochal distal stump on MRCP. PMID- 14565092 TI - [Experience in performance of percutaneous endoscopic gastrostomy in 60 children]. AB - The Percutaneous endoscopic gastrostomy (PEG) has radically changed the handling of the children with nutritional problems that until then were forced to receive nutritionals contributions by parenteral or enteral way, by nasogastric tube, conventional surgical gastrostomy or by central venous access. The objective of this work was to evaluate our experience with 60 patients, with diverse pathologies, that needed a PEG so that they could receive a suitable nutritional contribution. Were registered data of age, sex, reason for the accomplishment of the PEG, used surgical time, days of hospital stay and we analyzed the complications derived from the technique. There were two important complications. In one case one was a patient who presented a gastrocolic fistula as a result of the perforation of the colon in the accomplishment of the PEG and in another case a necrotizing fascitis in a girl who presented a choanal bilateral atresia. The rest of the complications were minor and they were treated without problems. The complications have been comparable to the registered in other series. Although the technique is simple and fast, is not free of risks, reason why it is necessary to inform suitably to the parents of the possibility of complications. In general, the degree of satisfaction of the parents was high. PMID- 14565093 TI - [Lung cystic adenomatoid malformation: our experience]. AB - BACKGROUND: Cystic Adenomatoid Malformation (CAM) is a rare entity in pediatrics. To know about it will lead us to its analysis, diagnosis and adequate surgical treatment. AIM: To show our experience for the diagnosis and treatment of lung (CAM). MATERIAL AND METHODS: Over a period from 1995 to 2000 twenty-one patients with (CAM) diagnosis were treated. Diagnosis was based upon clinical evaluation, laboratory and radiological findings: these include thorax X-ray, CT scan, echo Doppler and Angiography. The elective treatment was lobectomy. RESULTS: Twenty one patients from the Children's Hospital "Sor Maria Ludovica" of La Plata city were retrospectively studied over a period from 1995 to 2000. Patient's average age was 3.4 years, fourteen were feminine and seven masculine. The study consisted of clinical evaluation and imaging diagnosis: thorax-x ray; CT scan, echo Doppler and Angiography. SURGICAL TREATMENT: lobectomy. Pleural drainage was left. No deaths were registered. Pathological studies confirmed the diagnosis and type of CAM. Follow up was indicated in all patients. CONCLUSIONS: The awareness of the present pathology on the part of pediatricians makes possible considerations for careful diagnostic studies and early patient referral. PMID- 14565094 TI - [Non-coordinated micturition syndrome mimicking posterior urethral valves in a male neonate]. AB - High-grade of vesicoureteral reflux (VUR) has been reported in association with prenatal diagnosis of abnormal bladder function in a male neonate with postpatally no anatomical urethral obstruction and bladder dysfunction. This study was designed to describe clinical presentation of this entity in male neonates, the urodynamic pattern, prognosis in terms of renal function and therapeutic management since birth, in our experience with four cases. MATERIALS AND METHODS: We reviewed the records of 4 male neonates diagnosed at birth of bilateral high-grade VUR (grade > or = 4) with prenatally diagnosed hydronephrosis; thus 8 refluxing renal units (RRU) were studied. All cases had renal failure at birth. In all cases dimercaptosuccinic acid (DMSA) renography was performed. All RRU had reflux nephropathy; 5 RRU had moderate impairment of renal function (20-40%). In 3 RUU was demonstrated by a severe decrease in renal function (10-20% in 2 RUU, and less than 10% in 1 RUU). Micturating cystouretrography (MCUG) excluded the presence of posterior urethral valves; however, in all cases a tightened bladder neck ("bladder neck impression") was present. Endoscopy was performed in one baby excluding posterior urethral valves. Follow up ranged from 2 to 5 years (mean 3.5). RESULTS: In all 4 cases underwent transient urinary diversion during their first month of life. Urodynamic study revealed a high-risk bladder with low compliance, a reduced functional bladder capacity and a high residual urine volume in all cases. All the patients was placed on antibiotic prophylaxis and oral oxybutinin chloride. Three patients underwent bilateral Cohen ureteral reimplantation. The other boy underwent left nephrectomy, right Politano ureteral reimplantation and ureterocystoplasty. In the 7 RRU postoperative MCUG revealed cessation of reflux. Currently therapy includes antibiotic prophylaxis in 4, oral oxybutinin in 4 and intermittent catheterisation in 2 patients. 3 patients have normal renal function. The other boy underwent re-diversion because of renal failure. CONCLUSIONS: Fetal severe bilateral reflux nephropathy is a clinical entity almost exclusively in male neonate mimicking hyper-pressure syndrome due to urethral obstruction, in terms of evolution (to chronification and renal failure) and treatment (that requires primordial management of bladder dysfunction). Absence of anatomical urethral obstruction and urodynamic pattern suggest functional obstruction of bladder neck periurethral sphincter complex in fetal life as a cause for this syndrome. For this reason we consider it as a clinical presentation of fetal non-coordinated voiding in male or "posterior urethral valves like syndrome". PMID- 14565095 TI - [Non-palpable testicle: joint review of pediatric endo-surgical group and its management by laparoscopy]. AB - We present the joint serie of 5 Pediatric Centers with 256 patients diagnosed of cryptorchidism with nonpalpable testis treated by laparoscopy. We have found that Ultrasound examination is not very useful. This entity affects more to the left side 60%. In near half of the cases 46% we have not found any testicle and in 54% it was carried out an orquidopexy by different methods. In 27% we made an orquidopexy by Fowler-Stephens technique with 86% of good results. We think that laparoscopy is the best way of diagnosis and treatment of this patients. PMID- 14565096 TI - [Activity of a pediatric intestinal transplantation program in Spain]. AB - AIM: To analyze the outcome of children with intestinal failure (IF) included as candidates for intestinal transplantation (IT). Patients, Eight out of 23 children with IF assessed since July 1997 met criteria for IT and were included on the waiting list. The causes of IT were Short Bowel Syndrome (SBS) in 6 and Microvillus Inclusion Disease (MID) in 2. The indication of IT were end stage liver disease (ESLD) in 5 (related to total parenteral nutrition administration, TPN), progressive hepatic fibrosis in 2 and loss of venous access in 1. The patients with ESLD were included for combined liver-small bowel transplantation (LSBT) and the remaining for isolated intestinal transplantation (IIT). RESULTS: Two children died waiting for LSBT, 4 patients are on the waiting list, 2 for LSBT and 2 for IIT (length of stay: 4-11 months). Two children were transplanted, one with IIT in a 2.5 years old boy with MID, and one with LSBT in a 22 months girl with SBS and ESLD. Both patients recovered intestinal function after transplantation and are a live (follow-up of 19 and 10 months respectively). The LSBT's patient is under treatment for postransplant lymphoproliferative disease (PTLD). CONCLUSIONS: The lack of suitable donors for the small children candidate to IT explains the long period of stay on the waiting list and the high pretransplant mortality. Two strategies are possible; early referral of children with IF to a transplant center and surgical techniques like ex vivi-hepatic reductions of the LSB graft. PMID- 14565097 TI - [Epidermoid cyst of the mouth floor]. AB - The epidermoid cysts are frequent during childhood, however mouth floor location are very unusual, because of their more difficult diagnosis and therapeutic approach. We present a 5 years old male, symptoms free until a week before, when his parents noticed a well defined mass in the mouth floor. A physical examination leaded to the diagnosis of possible epidermoid cyst. The tumor was excised through an introral approach. A review of different diagnostic means and surgical management are undertaken. PMID- 14565098 TI - [Intestinal myopathy in Steinert's disease]. AB - The myothonic dystrophy or Steinert's disease is a congenital, autosomal, dominant disorder which seriously affects the striated muscle and also to a certain extent, several organs and systems and on rare occasions, the intestinal smooth muscle. In the case, we treated a four years old girl in whom Steinert's disease was diagnosed when she was born and who developed the characteristics of severe constipation after a few months of life. The ano-rectal manometry showed a paradoxical reaction of external Sphincter with a normal inhibitory reflex. The rectal biopsy revelated a miophathy which affected the muscularis propia with the normal neuronal innervation. The X-ray studies showed that motility disorder was stopped in the distal area of the left colon. A colostomy in the healthy zone worked extremely well. Six months later, the normal colon was brought down retro rectal. Only a partial incontinence remained in the external sphincter caused by Steinert's disease. The interest of this case lies in the pathological association, striade muscle and smooth muscle, in such an early age of life. It is a new contribution to the complex chapter of the intestinal pseudo obstruction. PMID- 14565099 TI - [Acute abdomen secondary to intra-abdominal hibernoma]. AB - The hibernoma is a rare benign tumor derived of embrionary fat seen in adult, which is usually localized in scapular and interscapular region. It express as a slowing growth bulk without symptoms. The authors present a 2 years old boy with acute abdomen requiring urgent laparotomy, that shows a tumor connected to mesocolon by a vascular pedicle that had undergone torsion. No other similar case published been found. The treatment is surgical resection although recurrences have been reported due to incomplete resection. PMID- 14565100 TI - [Pediatric surgery]. PMID- 14565101 TI - The HSJ interview: Professor Louis Appleby. Interview by Paul Smith. PMID- 14565102 TI - Clinical management where medicine meets management. Feelings of anxiety. AB - The psychiatrist vacancy rate means job redesign is inevitable if the mental health national service framework is to be achieved. The issue of patient responsibility is causing concern among some clinicians. Avon and Wiltshire Mental Health Partnership trust is making progress in changing referral system. PMID- 14565103 TI - Mental health. Sentence structure. AB - Mental health issues are endemic in prisoner populations, with many trusts struggling to provide specialist intervention. Before the inreach team at Bullwood Hall prison was put in place, most of its work was left to non specialist healthcare workers. PMID- 14565104 TI - Communications. Bang to writes. PMID- 14565105 TI - Finance. Loan ranger. PMID- 14565106 TI - HSJ people. World view. PMID- 14565107 TI - Recruitment and retention. Foreign legion. PMID- 14565108 TI - Recruitment and retention. Minority report. PMID- 14565109 TI - Recruitment and retention. Age-old solution. PMID- 14565110 TI - [On the centenary of the birth of Ivan Ivanovich Elkin]. PMID- 14565111 TI - [Nizhnii Novgorod period in the life of I. I. Elkin]. PMID- 14565112 TI - [I. I. Elkin's activity on the USSR People's Commissariat of Health]. PMID- 14565113 TI - [I. I. Elkin in my memory]. PMID- 14565114 TI - [Ivan Ivanovich Elkin--an outstanding person, teacher, scientist, public health organizer]. PMID- 14565115 TI - [About my teacher, with gratitude]. PMID- 14565116 TI - [Remembering Ivan Ivanovich Elkin]. PMID- 14565117 TI - [Main trends and problems connected with the improvement of epidemiology training of graduates of preventive medicine departments]. PMID- 14565118 TI - [How to reform the teaching epidemiology at a higher medical institution]. PMID- 14565119 TI - [Teaching experience in the field of management of epidemic control activity at the chair of epidemiology]. PMID- 14565120 TI - [Teaching experience on the problems of immunoprophylaxis at the chair of epidemiology]. PMID- 14565121 TI - [Role of epidemiological science and practice in protecting the health of the population of the country]. AB - The historical stages of the development of epidemiology as a science with a specific object of study, is summarized. One of the greatest theoretical generalizations is the doctrine on the natural focality of diseases. New principles of theoretical epidemiology, new theories and concepts are analyzed. The role of infectious pathology as one of the main criteria of the population health is emphasized as well as a wide spread of infectious (parasitic) diseases. The specific feature of the present stage of epidemiological situation is the dominating and constantly increasing role of viruses in the etiology of infectious diseases. The effectiveness of vaccinal prophylaxis to control vaccine preventable infections is substantiated. The role of epidemiological science in revealing the epidemiological regularities of emerging and re-emerging infections is emphasized. PMID- 14565122 TI - [Features of epidemiological activity in medicine]. AB - The article deals with the main elements of scientific knowledge in epidemiology, viz. its subject matter, goals and methods. The statement is substantiated that the main subject of epidemiology, determining its specificity, is the morbidity of the population, and the content of the epidemiological goals and methods are disclosed. The determination of the term "epidemiological approach" as well as a scheme reflecting the structure of epidemiology are presented. PMID- 14565123 TI - [Morbidity: terminology, mechanisms of formation and classification]. AB - In the 70-ies of the last century the evolution of the epidemic process doctrine resulted in developing population approach to morbidity analysis, in addition to the traditional focal approach. On the one hand, this led to the discussion on the possibility of applying such notion to non-infectious diseases and, on the other hand, to the discussion on the mechanisms of the development and formation of different manifestations in infectious diseases. In both cases the discussion was centered on the phenomenon of morbidity. The present article deals with the nature of morbidity as a phenomenon and the acceptability of general scientific approaches in the interpretation of seasonal and cyclic character of morbidity from the viewpoint of the wave process in an open system. On the basis of the data presented in this work the terms used in epidemiology in considering manifestations of the epidemic process are discussed. The improved classification of morbidity by qualitative and quantitative signs for infectious diseases is proposed. PMID- 14565124 TI - [Molecular epidemiology of infectious diseases]. AB - Data on molecular epidemiology of bacterial infections is summarized. The term definitions of "molecular epidemiology", "taxonomic species" are given, the limits and species structure of prokaryotes are described. The basic mechanisms of the prokaryotes variability in the epidemic process, as well as the possibilities and limitations of microbiological, molecular-biological and population--genetic typing methods, are characterized. The tactics of molecular biological studies in analyzing the population structure on the global, regional and local levels is presented. The economic effectiveness of measures taken with due consideration of information on the clonal structure of causative agents of hospital infections is shown. PMID- 14565126 TI - [Significance of epidemiological analysis for the development of epidemiology]. AB - The significance of epidemiological analysis for the study of the epidemic process is substantiated. The historical stages of the development of the doctrine on the epidemic process along with the role of epidemiological analysis in social and hygienic monitoring and epidemiological surveillance from the viewpoint of systemic approach are presented. The conclusion has been made on the necessity of the development of the systemic methodology of controlling the epidemic process and the sanitary and epidemiological situation. PMID- 14565125 TI - [Doctrine of the transmission of the causative agents of infections and social and ecologic concepts of the epidemic process]. AB - The concept on social and ecologic determination of the epidemic process in its direct connection with L.V. Gromashevsky's doctrine on the transmission mechanism of the causative agents of infection is presented. The role of this concept in the development of epidemiological science is discussed. PMID- 14565127 TI - ["Epidemic" vs "epidemiological": linguistic aspects of epidemiological terminology]. AB - The meaning of the terms "epidemic" and "epidemiological" used in a medical discourse have been analyzed; differential signs have been found by the method of linguistic analysis, making it possible to avoid unjustified interchanges of these terms. PMID- 14565128 TI - [Once again on the professional language of epidemiologists]. AB - Grounds for the adoption of a unified, commonly accessible, professional terminology by specialists are presented. The emphasis is made on the unification and uniformity of using the terms "epidemiological" and "epidemic". A small dictionary explaining the true or false sense of terms in epidemiology is proposed. PMID- 14565129 TI - [Epidemiological pattern of scarlet fever in recent years]. AB - During the period of 1953-2001 scarlet fever morbidity level fluctuated from 670.3 to 65.9 per 100,000 of the population in Moscow and from 531.9 to 35.0 per 100,000 of the population of the Russian Federation. In recent years an increased morbidity was more pronounced in Moscow than in the Russian Federation as a whole. Children formed the greater part of scarlet fever patients, the cases of scarlet fever among children in Moscow occurring more often than, on the average, in Russia. As before, annual morbidity among children attending children's institutions was higher 3- to 4-fold than among children brought up at home. This difference was most sharply pronounced among young children during the first two years of their life. In contrast to morbidity observed during previous 20-30 years, a drop in morbidity among children during the first two years of their life was registered, while morbidity level among children aged 3-6 years and 7-14 years increased. Scarlet fever morbidity had a pronounced seasonal (autumn winter) pattern. In a group of children aged 3-5 years who attended organized groups, on the average, 78.6% of scarlet fever cases fell on seasonal morbidity, the most prolonged one. PMID- 14565130 TI - [Mode of the rational use of Pseudomonas aeruginosa bacteriophages in therapeutic and epidemic control practice]. AB - Ecological aspects of the circulation of P. aeruginosa and P. aeruginosa bacteriophages under hospital conditions were under study. The statement concerning the formation of triple parasitic systems was put forward. The influence of these systems on the formation of phage and antibiotic resistance in P. aeruginosa hospital strains was studied. Spontaneous circulation of faintly virulent phages taking part in the formation of triple parasitic systems was shown not to ensure the elimination of P. aeruginosa hospital strains in clinics. Construction of highly virulent phages adapted to local P. aeruginosa strains was the only way of ensuring the protection of patients. Theoretical and practical approaches to the use of highly active bacteriophages for controlling P. aeruginosa infection were substantiated. The realization of these approaches resulted in achieving not only a clinical, but also essential epidemic control effect in cases of purulent septic infections caused by P. aeruginosa (a decreased frequentcy of hospital infections from 40.8% to 8.93%). PMID- 14565131 TI - [Epidemiological survey of a single-case focus of acute enteric infections]. AB - The aim, tasks and stages of the epidemiological survey of a single-cased focus of infection, taking into account present-day specific features of the epidemiology of acute enteric infections, have been formulated. The technical approaches to the detection of the source, routes, factors and conditions of the causative agent transmission are given. The patterns of the conclusion on the epidemiological survey of the focus are given. PMID- 14565132 TI - [Methodological aspects of the evaluation of epidemiological safety in maternity hospitals]. AB - The distribution of morbidity and mortality in newborns, as well as morbidity of parturient women, in maternity hospitals of Moscow official have been analyzed according to statistical data for 1996-1999. The methods of cluster analysis (k medium and tree classification by the method of the next door neighbours) were used. The evaluation of the stability of the distribution of morbidity and mortality in maternity hospitals was made with the use of chi 2 criterion. The specific features of the distribution of morbidity and mortality of newborns, as well as morbidity of parturient women, in the hospitals under study were detected. The methodological approach to the evaluation of epidemiological safety in maternity hospitals was proposed. PMID- 14565133 TI - ["Quorum sensing" or social behavior of bacteria]. AB - The review deals with the data of literature on the role of the "quorum sensing" (QS) system ensuring the social behavior of bacteria in the regulation of virulence genes. The mechanisms of the action of these systems in Gram-negative and Gram-positive bacteria, as well as the influence of acyl-homoserine lactones, one of the components of the QS system, on the immune response of the infected host are discussed. In addition, in this review the data of literature on the existence of bacteria in the form of biofilms are presented. The methods of the identification of biofilms, the methods of their experimental preparation and the role of the QS system in the process of their formation are considered. PMID- 14565134 TI - [Current epidemiological surveillance of tuberculosis]. AB - Some present-day specific epidemiological, microbiological and clinical features of tuberculosis are presented. A variant of the epidemiological surveillance morbidity scheme, comprising 3 blocks, viz. information, analysis and management, is proposed. In the information block the basic data necessary for epidemic analysis is listed, while the diagnostic block contains the stage-by-stage analysis of the information block data. In the block of management decisions the main directions of the tuberculosis prophylaxis interventions are listed. PMID- 14565135 TI - [Some unsolved aspects of the parasitism problem]. AB - Various interpretations of the parasitism phenomenon exist. In this work data supporting the ecological understanding of its nature are presented. For parasitic species formed in the process of evolution the host organism has become the habitat (nutrition, multiplication), i.e., the environment, where interactions between the host and parasites are governed by ecological regularities. The consequences of this interactions are different (disease, asymptomatic infection), being secondary they reflect the result of concrete interactions. For this reason parasitism should not be identified in terms of only one consequence of such interaction--the development of pathologic processes. PMID- 14565136 TI - [Specific prophylaxis of shigellosis]. AB - A high level of shigellosis morbidity requires new approaches to the control of bacterial dysentery. According to modern concepts, the outbreaks of Shigella infections are linked both with less intensive epidemic control measures and the objective cyclic character of the epidemic process. In this connection great importance is attached to the development of vaccines for the immunization of high risk groups in territories with unfavorable epidemic conditions and in zones of military conflicts, as well as children of school and pre-school age, who mostly determine seasonal rises of shigellosis morbidity. In this article the data describing new approaches to the creation of new live enteric vaccines on the basis of the knowledge of the genetic control of microbial pathogenicity and the regulation of its expression are presented. Attenuated Shigella mutants, created by different authors and having good prospects to be used for the development of vaccines, are characterized. PMID- 14565137 TI - [Isolation and identification of the infective agent of severe acute respiratory syndrome (SARS) from a patient with atypical pneumonia]. AB - The virological, morphological, molecular biological and immunochemical study of the infective agent isolated from the patient with the symptoms of atypical pneumonia, hospitalized in the infectious department of the clinical hospital in Blagoveshchensk, was carried out. Thus the fact of the appearance of SARS virus on the territory of Russia was proved. The isolated infective agent, identified as coronavirus strain CoD, was partly characterized and deposited to the virus collection of the Center of Special Laboratory Diagnostics and Treatment of Quarantine and Exotic Infectious Diseases. PMID- 14565138 TI - [Human cases of hydrophobia in the Russian Federation]. AB - The analysis of 29 human cases of hydrophobia registered in the Russian Federation in 2000-2001 is presented. Dogs appeared to be the main infection source (50% of cases). An increased proportion of cats (17.3%) in the spread of the disease was noted. Among wild animals, the leading role of foxes in the spread of rabies virus was determined. A number of serious drawbacks in the organization of antirable service to the population, in particular, insufficient qualification of medical personnel, a low level of medical education among the population on the sources of infection, deviations from the rules of urgent notification presentation, etc. The conclusion was made on the necessity of improving the organization of antirabic services to the population. PMID- 14565139 TI - [Types of immune response in acute infectious diseases]. AB - The author's concept on the system of self regulation of immune response is described. Four types of this response are proposed, which differ by total intensity of cytokine reaction, the development time as well as manifestation degree of non specific immunosuppression, and, most importantly, the profile of specific immune response to the antigens of the infective agent. These four types of immune response are closely linked with increased severity of clinical manifestations. PMID- 14565140 TI - [Acidic stress in bacteria]. AB - Information on acidic stress in bacteria, studied not only on the phenomenological, but also molecular and genetic levels, are systematized. Acidic stress in bacteria, appearing as the result of the acidification of the medium, is characterized by many events on the level of gene regulation. An increased expression of some genes and a decreased expression of others result in growth deceleration, quantitative and qualitative changes in the synthesis of proteins. Some of the newly synthesized proteins ensure the survival of bacteria in a medium with higher acidity and their protection from other stresses. The applied importance of acidic stress is relevant to some aspects of biotechnology, immunobiology and medicine. PMID- 14565141 TI - Born free. PMID- 14565142 TI - Varsity challenge. PMID- 14565143 TI - The heat is on. PMID- 14565144 TI - Prize fight. PMID- 14565145 TI - The HSJ interview: Harry Cayton. Hearing voices. Interview by Alastair McLellan. PMID- 14565146 TI - Clinical management where medicine meets management. The American way. AB - Close relationships between business managers and clinicians, and purchaser and provider, are valuable in chronic-disease management. Organisations used high quality clinical information and performance management. Chronic-disease management based on risk-management, proactive case managers and patient education is effective. PMID- 14565147 TI - US special. States of change. PMID- 14565148 TI - Law. On duty. PMID- 14565149 TI - HSJ people. Let's get physio. PMID- 14565150 TI - Buildings. Try this for size. PMID- 14565151 TI - Buildings. Cash dash. PMID- 14565152 TI - Buildings. Greenfield, red light. PMID- 14565153 TI - Buildings. Waste watchers. PMID- 14565154 TI - Planetarium Inversum -- a space vision for Earth education. AB - In a planetarium, the visitor is sitting on Earth and looking into an imaginary space. The Planetarium Inversum is the opposite: visitors are sitting in a space station, looking down on Mother Earth. It is a scientifically-based information show with visitors involvement, its elements being partially virtual (Earth in space has to be projected with highest possible resolution) but also containing real structures, such as the visitors' Earth observatory with adjacent biological systems (plant cultures and other ecological life support components). Its main message concerns the limits and the vulnerability of our home planet, its uniqueness, beauty and above all, its irreplaceableness: Earth does not have an emergency exit. The Earth observatory is part of a ring shaped, rotating space station of the type designed by Wernher von Braun decades ago. Visitors are told that gravity is being substituted by centrifugal force. Both types of life support systems are being demonstrated--self regenerative life based ones and technical ones as a backup (solar electric splitting of water and chemical absorption of respiratory CO2). PMID- 14565157 TI - Dad. PMID- 14565156 TI - Society of Biomaterials Graduate Degree Candidate Student Research Award. Matrix metalloproteinase inhibitor within an absorbable coating for vascular applications: delivery device characterization and reduction of smooth muscle cell proliferation and migration. AB - Significant occurrences of arterial restenosis remain a complicating factor of endovascular stent implantation. The incorporation of GM6001, a matrix metalloproteinase inhibitor (MMPI), into a poly(lactide-co-glycolide) (PLGA) absorbable coating for 316L stainless steel is proposed as a means to reduce the rate of smooth muscle cell proliferation and migration. Coatings were fabricated using a solvent evaporation technique, and the surfaces were characterized by noncontacting profilometry and scanning electron microscopy. Sufficient degradation of the PLGA determined by gel permeation chromatography occurred to release adequate amounts of the GM6001 from the coating within a 7-day period. A cumulative GM6001 release at day 42 was determined to be 77.8 +/- 1.4% of the actual GM6001 content within the coating. The coating containing the GM6001 reduced the rate of in vitro cell growth of human aortic smooth muscle cell (HASMC) by 30.7 and 37.4% compared to the metallic substrate only after 4 and 7 days, respectively. However, the MMP-2 activity normalized to cell number was not statistically different between the GM6001 releasing coating and the metal substrate. Using a scrape wound injury assay, the migration of HASMCs was shown to be decreased by 21.4% with GM6001 released from the PLGA coating compared to metallic substrate only. These results suggest that releasing a MMPI from an absorbable coating of a metallic substrate provides a reduction of HASMC proliferation and migration rates, while preserving the overall MMP activity in efforts to retain normal cellular regulation. PMID- 14565158 TI - Preliminary observations on genetic alterations in pilocytic astrocytomas associated with neurofibromatosis 1. AB - Neurofibromatosis 1 (NF1) is an autosomal dominant disorder that predisposes sufferers to various forms of neoplasia. Among affected individuals, 15%-20% develop astrocytomas, especially pilocytic astrocytomas (PA), which are benign and classified as grade I by the World Health Organization. They are generally well circumscribed, and their progression is slow. NF1-associated PAs (NF1-PAs) occasionally behave as aggressive tumors. To elucidate underlying genetic events in clinically progressive NF1-PAs, we performed molecular genetic analysis on 12 PAs, including 3 NF1-PAs, for pS3, p16, and epidermal growth factor receptor genes, as well as loss of heterozygosity (LOH) on chromosome 1p, 10, 17, and 19q. None of the obvious genetic alterations typically seen in higher grade astrocytomas were found in 9 sporadic PAs. However, in 2 of 3 NF1-PAs, microsatellite analysis showed LOH10, including the PTEN (phosphatase and tensin homolog deleted on chromosome 10) gene locus, despite the diagnosis of pilocytic astrocytoma;one of these also manifested homozygous deletion of the p16 gene. The other NF1-PA harbored only LOH of the NF1 gene locus (17q). Our preliminary results support the hypothesis that some NF1-PAs differ genetically from sporadic PAs. PMID- 14565159 TI - Antiangiogenic effects of dexamethasone in 9L gliosarcoma assessed by MRI cerebral blood volume maps. AB - Depending on dose, dexamethasone has been shown to inhibit or stimulate growth of rat 9L gliosarcoma and decrease the expression of vascular endothelial growth factor (VEGF), an important mediator of tumor-associated angiogenesis. We demonstrate, by constructing relative cerebral blood volume (rCBV) maps with MRI, that dexamethasone also decreases total blood volume while increasing microvascular blood volume in Fischer rats bearing intracranial 9L gliosarcoma. Animals were inoculated with 1 x 10(5) 9L gliosarcoma tumor cells. On days 10-14 after tumor cell inoculation, animals were intra-peritoneally injected with dexamethasone (3 mg/kg) over 5 days. MRI-derived gradient echo (GE) and spin-echo (SE) rCBV maps were created to demonstrate total vasculature (GE) and microvasculature (SE). After MRI studies were performed, the rat's vasculature was perfused with a latex compound. Total vessel volume and diameters were assessed by microscopy. Dexamethasone decreased the tumor-enhancing area of postcontrast T1-weighted images (P < 0.0001) and total tumor volume(P = 0.0085). In addition, there was a greater than 50% decrease in GE rCBV (total vasculature) (P = 0.007) as well as a significant decrease in total fractional blood volume, as validated by histology (P = 0.0007). Conversely, there was an increase in SE rCBV signal (microvasculature) in animals treated with dexamethasone (P = 0.05), which was consistent with microscopy (P < 0.0001). These data demonstrate that (1) dexamethasone selectively treats tumor vasculature, suggesting a vessel-size selective effect and (2) MRI-derived rCBV is a noninvasive technique that can be used to evaluate changes in blood volume and vascular morphology. PMID- 14565160 TI - SPARC affects glioma cell growth differently when grown on brain ECM proteins in vitro under standard versus reduced-serum stress conditions. AB - Secreted protein acidic and rich in cysteine (SPARC) has a suppressive effect on U87 glioma cell proliferation when assessed in vitro and in vivo using parental U87T2 and U87T2-derived SPARC-transfected clones. Since SPARCinteracts with extracellular matrix (ECM) proteins, we examined the effect of SPARC secretion on proliferation, morphology, and cell density of glioma cells grown in vitro, in the absence and presence of ECM proteins under standard (10% fetal bovine serum [FBSI) and reduced (0.1% FBS) serum stress conditions. Under standard conditions, MTT (3-(4,5-cimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide) growth curves, morphology, and Western blot analyses demonstrated that SPARC had a suppressive and biphasic effect on growth that was not grossly modulated by the ECMs. The SPARC-induced changes in morphology observed at 24 h were not altered by the presence of ECMs. Under reduced-serum stress conditions, Western blot, morphological, and flow cytometric analyses indicated that the SPARC-induced suppressive growth effects were eliminated when the cells were grown on plastic. However, ECM-specific changes in growth were observed, some of which correlated with secreted SPARC levels. These results indicate that the differential effects of SPARC and ECMs on proliferation are dependent on culture conditions. Since the results obtained under standard conditions agree with our in vivo observations, we conclude that the ability of SPARC to suppress proliferation is regulated to a greater degree by the level of SPARC and that this suppressive effect is not influenced by the presence of any of the ECMs examined. PMID- 14565161 TI - A 45-year-old woman with fever and splenic infarcts. PMID- 14565162 TI - Does brain tumor histology influence cognitive function? AB - This retrospective study investigated the relationship between tumor histology and postsurgical cognitive function in patients diagnosed with malignant brain tumors. The neuropsychological functioning of 24 adult patients diagnosed with glioblastoma multiforme (GBM) was compared with the neuropsychological functioning of 24 adult patients diagnosed with anaplastic astrocytoma (AA). The groups were matched with respect to patient age, gender, and education, as well as tumor location and tumor volume. The mean raw test scores of the AA patient group were superior to the mean scores of the GBM patient group on nearly all measures administered. However, significant performance differences were not detected for any of 5 neuropsychological domain scores (Intellectual, Language, Memory, Executive, and Motor Function). Analysis of covariance (ANCOVA) revealed that tumor histology was not a significant predictor of domain score after controlling for tumor volume. Multiple regression and correlation analyses supported the results of the ANCOVA by offering further evidence of weak relationships between tumor type, tumor volume, and neuropsychological test scores. We conclude that tumor histology is not clearly predictive of cognitive performance in adults with AA and GBM. PMID- 14565164 TI - Importance of MR technique for stereotactic radiosurgery. AB - We investigated how frequently the imaging procedure we use immediately prior to radiosurgery--triple-dose gadolinium-enhanced MR performed with the patient immobilized in a nonrelocatable head frame and 1-mm-thick MPRAGE (magnetization prepared rapid gradient echo) images (SRS3xGado)-identifies previously unrecognized cerebral metastases in patients initially imaged by conventional MR with single-dose gadolinium (1xGado). Between July 1998 and July 2000, the diagnoses established for 47 patients who underwent radio-surgical procedures for treatment of cerebral metastases at The Gamma Knife Center of New York University were based initially on the 1xGado protocol. In July 1998, we began using SRS3xGado as our routine imaging protocol in preparation for targeting lesions for radio-surgery, using triple-dose gadolinium and acquisition of contiguous 1 mm Tl-weighted axial images. Because our SRS3xGado scans sometimes unexpectedly revealed additional metastases, we sought to learn how frequently the initial 1xGado scans would underestimate the number of metastases. We therefore reviewed the number of brain metastases identified on the SRS3xGado studies and compared the results to the number found by the 1xGado protocol, which had initially identified the brain metastases. Additional metastases, ranging from 1 to 23 lesions per patient, were identified on the SRS3xGado scan in 23 of 47 patients (49%). In 57% of the 23 patients, only one additional lesion was identified. The mean time interval between the 1xGado and the SRS3xGado scans was 20.6 days (range, 4-83 days), and the number of additional lesions detected and the time interval between two scans were negatively correlated (-0.11). The number of lesions detected on the SRS3xGado was associated only with the number of lesions on the 1xGado and not with any other patient or tumor pretreatment characteristics such as age, gender, largest tumor volume on the 1xGado, or number of days between the 1xGado and the SRS3xGado or prior surgery. The identification of additional lesions with SRS3xGado MR may have implications for patients who are treated with stereotactic radiosurgery alone (without whole brain irradiation) with single-dose gadolinium imaging, in that unidentified lesions may go untreated. As a result of these findings we continue to use and advocate SRS3xGado scans for radiosurgery. PMID- 14565163 TI - Phase 2 study of idarubicin in pediatric brain tumors: Pediatric Oncology Group study POG 9237. AB - Idarubicin (IDA), the 4-demethoxy analog of daunomycin, has had significant cytotoxicity in many malignancies. In previous reports, the alcohol metabolite of IDA, 4-demethoxydaunorubicinol (idarubicinol, or IDOL), had cytotoxic activity and the ability to penetrate the blood-brain barrier. For this reason, the Pediatric Oncology Group conducted a Phase 2 trial of IDA for children with recurrent or progressive brain tumors. Ninety-one eligible children were entered on this study, with ages ranging from 3 months to 19 years. Patients were stratified by tumor types into 6 categories: stratum 1, low-grade astrocytoma; stratum 2, malignant glioma (glioblastoma multiforme and anaplastic astrocytoma); stratum 3, medulloblastoma; stratum 4, brainstem glioma; stratum 5, ependymoma; and stratum 6, miscellaneous malignant tumors not included in the previous diagnoses. IDA(18 mg/m2) was infused over 4 h and followed by granulocyte colony stimulating factor (G-CSF) beginning day 5 after infusion of IDA. G-CSF was continued until blood cell count recovery. Cycles were repeated at approximately 21-day intervals until patients developed progressive disease or had completed 6 cycles with stable or improved disease. Pharmacokinetic plasma and cerebrospinal fluid (CSF) samples were collected from a subset of these patients. Response was poor in all strata. Most patients developed progressive disease; however, in 21 patients with medulloblastoma there was 1 partial response, and 6 patients had stable disease (SD) that in 4 patients lasted more than 20 weeks. Grades 3/4 hematopoietic toxicities were the most common toxic events, and 14 infection related events resulted in hospitalization of patients. Only 1 patient developed reduced cardiac function. The systemic clearance data for IDA and IDOL were nearly identical to those published on patients with leukemia, and the plasma elimination of the IDOL metabolite was substantially longer than that of the parent drug IDA. The peak CSF:plasma ratios of IDA and IDOL were very low. The overall response rates to IDA were disappointing despite periods of prolonged SD in nearly a fourth of the patients. We conclude from this data and from that in nonhuman primates that it is unlikely that IDA, daunomycin, or other related anthracyclines will be useful for treating primary CNS tumors. PMID- 14565165 TI - Inflammation 2003-Sixth World Congress. 2-6 August 2003, Vancouver, Canada. PMID- 14565166 TI - Inflammation 2003-Sixth World Congress. Nitric oxide and IBD. 2-6 August 2003, Vancouver, Canada. PMID- 14565167 TI - Inflammation 2003-Sixth World Congress. Focus on chronic obstructive pulmonary disease. 2-6 August 2003, Vancouver, Canada. PMID- 14565168 TI - Inflammation 2003-Sixth World Congress. Anti-inflammatory mechanisms. 2-6 August 2003, Vancouver, Canada. PMID- 14565169 TI - Metabolic Diseases Drug Discovery World Summit--SRI conference. 28-29 July 2003 San Diego, CA, USA. PMID- 14565170 TI - Metabolic Diseases Drug Discovery World Summit--SRI conference. Diabetes and obesity. 28-29 July 2003 San Diego, CA, USA. PMID- 14565171 TI - International Brain Research Organization--Sixth World Congress of Neuroscience. 10-15 July 2003, Prague, Czech Republic. PMID- 14565172 TI - Progressive vision loss in a man from Cameroon. PMID- 14565173 TI - 18th American Peptide Symposium. Peptide revolution: genomics, proteomics and therapeutics. 19-23 July 2003, Boston, MA, USA. PMID- 14565174 TI - International Society of Thrombosis and Hemostasis--XIXth Congress 12-18 July 2003, Birmingham, UK. PMID- 14565175 TI - Controlled Release Society--30th Annual Meeting and Exposition. 19-23 July 2003, Glasgow, Scotland, UK. PMID- 14565176 TI - HIV pathogenesis and treatment--second International AIDS Society conference. 13 16 July 2003, Paris, France. PMID- 14565178 TI - [Pietism and medicine in the first half of the 18th century]. AB - In the beginning of the 18th century Halle Pietists tried to establish a specific approach to sickness and healing. They constructed close links between physical illness on the one hand and the religious concept of individual piety, penance and rebirth on the other. This new' pietist medicine largely depended on Georg Ernst Stahl's medical theory, which was not pietist in itself, but was adopted and simplified by pietist physicians. Although conclusive and rhetorically present in programmatic texts, pietist medicine turned out to be less influential on medical practice than expected. PMID- 14565177 TI - [Pharmacy as a means of propagating the Lutherian doctrine of justification]. AB - Pharmacy serving to propagate the Lutherian doctrine of justification: "Christus as a pharmacist" is an interconfessional, but confessionally differentiated symbolic motif (Sinnbildmotiv) of Christian folklore art in German-speaking countries. The article investigates the sociocultural conditions and prerequisites (German bible translation, religion and confession, piety, pharmacy, chymiatry, chemistry, apothecary training and status) for transfering the old metaphor and idea of Christ as a physician to the new vision of Christ as a apothecary who prepares and dispenses his heavenly medicine all by himself. In the early 17th century (especially in the 1610's) these requirements were fulfilled, so that the oldest known witness to this motif transfer (picturing the so-called Heilandsruf of Matthew 11, 28), a genre picture of 1619, will be the first pictural version of this motif in general. It was created by the protestant Painter Mich(a)el Herr of Nuremberg. In the abstract and reduced form of a devotional picture this motif then became widespread in churches and vicarages, in monasteries and their apothecaries as well as in private houses (with small altars: Herrgottswinkel). The oldest yet known examples are works from around 1630. For the first time during the Thirty Year's War, it served in this form for propagating the Lutherian justification doctrine (now referring to Jesaja 55, 1), saying that the belief in Christ is enough to be released from all sins (sola fide). Around 1650, as a reaction to that, a catholic version of the devotional picture was created, claiming and picturing that in contrast eucharist is the highest and real, healing medium' of Christ to redemption. All pictures of this version avoid quoting Jesaja 55, 1, whereas all protestant pictures quote this verse from the Bible word-for-word. PMID- 14565179 TI - Professor David Poswillo, CBE, 1927-2003. PMID- 14565180 TI - Denis Parsons Burkitt (1911-1993). PMID- 14565181 TI - Discovery of the first local anaesthetic--Carl Koller (1857-1944). PMID- 14565182 TI - The Caparn family. PMID- 14565183 TI - The Edinburgh apothecaries. PMID- 14565184 TI - Aldred Scott Warthin (1866-1931). PMID- 14565185 TI - Metal boxes for cough lozenges. PMID- 14565186 TI - [Birth and infant abandonment at the Port-Royal hospital at the beginning of the 19th century]. PMID- 14565187 TI - [The surgical school of the port of Rochefort (1722-1789): a model of the old regime]. PMID- 14565188 TI - [English medicine in the Tudor times]. AB - In the Tudor times in England medicine was influenced by Renaissance and ideas of humanism. Medical humanists were searching for and discovering genuine ancient manuscripts in the interest of establishing a "pure" classical terminology and achieving an elegant Latin style uncontaminated by Arab intermediaries. The theory of disease, diagnosis and treatment were all dominated by Galen and his theory of humours. Advances in anatomy and physiology did not bring about any change to the status quo. Apart from university-trained doctors, medical skills were offered by surgeons, apothecaries and various quacks, empirics and mountebanks. There was a selection of medical books translated into and written in the vernacular English they could choose from, apart from books written in Latin and translated into it. PMID- 14565190 TI - [Psychiatry in 19th century Cracow (1820-1880)]. AB - This paper contains information about Cracovian mental clinics treating mentally ill people, about the people treated in the psychiatric wards and their doctors in the years 1820-1880. Psychiatric institutions changed gradually - initially, the ill were placed and treated in an old medieval building, previously serving as a monastery, and in the final period, in a newly built complex originally intended for a psychiatric ward. Gradually,the specialization of psychiatrist was developed - it was to be a medical specialist dealing above all with mentally ill patients. The community of Cracovian psychiatrists contributed a lot to the development of specialist Polish terminology describing mental disorders. I have presented some important papers devoted to the aspects of psychiatry written by doctors directly dealing with the mentally ill and also by the doctors of different specialisations, mainly from the field of forensic medicine. In the years discussed, there were attempts to put psychiatry on a par with different fields of medicine by means of changing the conditions of treating the ill and introducing lectures on psychiatry in the Medicine Faculty of the Jagiellonian University. PMID- 14565189 TI - [History of the barber-surgery in Rzeszow]. AB - The beginning of barber-surgeons activity in the Rzeszow area was presented in this paper. The first entry of the presence of barber-surgeons in Rzeszow dates from 1484 and informs about a conflict between them and the bath attendants. In 1626, following the example of other Polish cities, the first barber-surgeons guild was established also in Rzeszow. The names of its founders are cited in this paper. Albert Boniecki was the first head of the surgical guild. In consequence of changes in the composition of the population and religious faith, a new guild was created in 1699, whose members were both, Christian and Jews. Until to the end of the so called First Polish Republic, the surgical activity in Rzeszow was in the hands of guild-educated barber-surgeons. Two tables give the numbers and names of Christian and Jewish surgeons in particular years. Figures bring the shape of original documents. PMID- 14565191 TI - [How to set up a family and have healthy children? Plato and Aristotle's formulas for good life]. AB - The article reconstructs and analyses the views of two great philosophers: Plato and Aristotle. Contrary to general opinions, philosophy busies itself not only with abstract issues, but also with the realistic ones. The thinkers introduced here prove this theory. They went into details discussing matters such as: procreation, behaviour of pregnant women, feeding and bringing up babies, children and teenagers' physical and mental education. Both men treated citizens' health as a matter of paramount importance, the matter of national significance. PMID- 14565192 TI - [First research work by Robert Koch on etiology of anthrax-in cooperation with Jozef Knechtel, Polish apothecary]. AB - Terroristic attack on United States of America 11 September 2001 and just after many cases of anthrax spores employment as biological warfare called our attention to Robert Koch. He determined anthrax etiology and enclosed it in his first research work: Die Aetiologie der Milzbrand-Kranheit begrundet auf die Entwicklungsgeschichte des Bacillus Anthracis. The results of this research are widely described. In the scientific researches participated J. Knechtel, Pole, pharmacist, pharmacy owner in Wolsztyn. His adjacent laboratory near pharmacy was provided with microscope, camera, table and two chairs. Many slides and above mentioned article / without J. Knechtel as joint author/were the results of this findings. About cooperation Pole with R. Koch we found out from two letters doctor Brinkmann' s authorship and three reports explored by A. Skrobacki in Central Register Office in Merseburg. The objects mentioned above were delivered by J. Knechtel's widow as the gift to Institute of Infectious Diseases in Berlin in 1905. Robert Koch' s cooperation with a Polish pharmacist was concealed. It was caused by a historic background and the policy of Prussia - an invader state in relation to Polish people. The official demonstration of cooperation with a Polish pharmacist under these circumstances could not take place. PMID- 14565193 TI - [Outline biography of a forgotten Polish doctor from Estonia]. AB - The paper presents the figure of Polish doctor Wladyslaw Wawrzyniec Mierzejewski who was born in 1841 in the province of Witebsk. He studied medicine in Petersburg. After graduation in the field of gynecology followed by the achievement of MD, he moved to the city of Kuressaare on the Saaremaa Island in 1876. That year he established a natural therapy clinic there and treated female patients from Petersburg, Tallin and Dorpat. The clinic was taken over by his son Gonzaga George Mierzejewski, a gynecologist himself. The clinic was destroyed during II World War in 1941. At present, the former location of the balneology clinic is commemorated with a stone with a plate for the memory of Polish doctors. PMID- 14565195 TI - Bibliography. Current world literature. Psychiatry. PMID- 14565194 TI - Bibliography. Current world literature. Cardiovascular medicine. PMID- 14565196 TI - Bibliography. Current world literature. General pediatrics. PMID- 14565197 TI - [A study about Il Chun Yu]. AB - Il Chun Yu was the first Korean microbiologist and the Korean student who studied in Germany. He got the two doctor degree: one in Freiburg University of Germany in 1924, and the other Keio University of Japan in 1926. He became a professor of Kyungsung Medical Junior College in 1926, and as the first grade scholar in the Korean microbiology field. He issued many essays in several mass communication media to cultivate people and played an important role in the field of health hygiene by contributing to the improvement of health hygiene. PMID- 14565199 TI - [Cultural hegemony of medicine in modern China]. AB - The paper is to explore into how cultural hegemony had been established in modern China, focused on ideological debates and political conflicts between modernists and traditionalists. Relying upon historical, anthropological, and medico historical researches respectively by Paul Cohen, Judith Farquhar and Paul Unschuld, I criticize free research paradigms that had prevailed in modern Chinese History: (i)the 'Chinese response to Western impact' perspective fails to explain how Chinese Western medical practitioners founded their own independent organization; (ii) a dichotomy of 'tradition versus modernity' is, from an epistemological viewpoint, incompatible with an ontological view of illness shared between traditional Chinese medicine and Western medicine; and (iii) while those Weberian social scientists tend to consider culture as the system of meanings and symbols, separated from their temporal and spatial matrix, they neglect political and historical spheres that are inevitably represented in cultural hegemony. My arguments are divided into two parts. The first part investigates that whereas Chinese modernists aggressively supported an immediate institutionalization of Western medicine for getting adapted to social Darwinian world, neo- traditionalists tried to maintain medical identity through national essence backed up by Chinese civilization. In the second part, the paper illuminates how having emerged as a conceptual idea for moving beyond 'tradition versus modernity', 'state medicine' became popularized to solve public health problems in 1930s' rural China. In conclusion, cultural hegemony-oriented debates that were seriously staged in the 1920s and 1930s between modernists and neo traditionalists were transformed to "scientification of traditional Chinese medicine and popularization of Western medicine" a slogan proposed by Mao Ze Dong. PMID- 14565200 TI - Attracting nurses. PMID- 14565201 TI - Solutions for inadequate staffing. PMID- 14565203 TI - End of an era for nurses' clinic. PMID- 14565202 TI - [The making of hygienic modernity in Meiji Japan, 1868-1905]. AB - This article is based on conceptual and methodological understanding of hygienic modernity in the nineteenth-century Western countries: one is the concept of modern hygiene in the context of modern state and the other is methodological relation of modern hygiene to scientific theory of germ . While modern state calls for the institutionalization of medical police as an administrative tool for consolidating the governmentality what Michel Foucault calls, scientific 'invention' of germ may be considered as 'logical, philosophical and historiographical'. Furthermore, the Meiji medicine men preferred Koch's to Pasteur's laboratory framework, not because the former was scientific than the latter but because Koch's programs were more compatible with imperial needs. The objective of this paper is to investigate four ways in which hygienic modernity had been established in Meiji Japan; (i) how Meiji imperialists perceived and managed to control Japanese hygienic condition, (ii) how Meiji-leading doctors learned about the German modern system of hygiene to consolidate Meiji empire; (iii) how modern germ theory functioned as the formation of imperial bodies in Meiji period; and (iv) how modem military hygiene contributed to Japanese defeat of Russia. Although I try to contend that modern hygiene was adopted as one of the most significant strategies for intensifying and extending the Meiji empire, this paper has some limits in not identifying how Japanese perception of infectious diseases were culturally adaptive to science-based hygienic programs the Meiji administrators had installed. PMID- 14565204 TI - Geriatric nurse practitioner care guidelines: pruritus and xerosis in the elderly person. PMID- 14565205 TI - [Absorption of plasticizer probably higher than expected]. PMID- 14565207 TI - [Further indications for tenofovir]. PMID- 14565206 TI - [The concept of disease in Galen]. AB - Galen was, with no doubt, a great authority in ancient medicine rivalled only with "the father of medicine" Hippocrates. His medicine inherited not only Hippocratic tradition, which is characterized by dynamic pathology, but also Alexandrian medicine, which made a great contribution to anatomy. He did not generalize all the pathological phenomena according to one dogmatic theory. His medicine was open to all kinds of medicine, but he was quite selective in receiving different medical traditions.Galen defined disease as impairment of bodily activities. Whatever that impairs the bodily activities is the cause of disease. Galen' s pathology is built upon two heterogeneous medical traditions: 1) Dynamic pathology of the Hippocratic medicine, but which ignored anatomy 2) Anatomical pathology of Alexandrian medicine. Galen integrated these two different traditions by his concept of disease. His definition of disease, impairment of bodily activities, made it possible to harmorize these two different traditions otherwise which would have been conflictual. It is Galen' s great contribution to Western medicine to have laid a foundation of pathology by combining physiological and anatomical point of view . PMID- 14565208 TI - [Atazanavir--first protease inhibitor for one dose administration]. PMID- 14565209 TI - On treating sexuality. PMID- 14565210 TI - William Stewart Halsted in the history of American surgery. PMID- 14565211 TI - On treating sexuality. PMID- 14565213 TI - On treating sexuality. PMID- 14565214 TI - On treating sexuality. PMID- 14565217 TI - Fraud and deception in science. PMID- 14565216 TI - Adult respiratory infections. PMID- 14565218 TI - HIV-1 Vif: counteracting innate antiretroviral defenses. PMID- 14565219 TI - Large animal models: bridging the gap. PMID- 14565221 TI - Calcium intake and prostate cancer risk in a long-term aging study: The Baltimore Logitudinal Study of Aging. PMID- 14565220 TI - Adrenocortical steroidogenesis and aminoglutethimide I. Biomedical studies. PMID- 14565222 TI - Screening for prostate cancer: an update of the evidence for the U.S. Preventive Services Task Force. PMID- 14565223 TI - Screening for prostate cancer: recommendation and rationale. PMID- 14565224 TI - [Crohn disease]. PMID- 14565225 TI - Oligonucleotides and derivatives as gene-specific control agents. AB - The current achievement of genome sequence projects of a dozen eukaryote organisms (including human genome) and the development of functional genomics are providing the basic knowledge required to utilize gene-specific reagents for both basic understanding of cell physiology and therapeutical development. The field of chemical genomics has the ambitious goal of designing molecules that could act selectively on every single gene or gene product in a cell and in vivo. The progress in oligonucleotide-based approaches will be the topic of this review, however, other nucleic acid- and SELEX-based approaches as well as high sequence specific low molecular weight DNA-specific ligands will also be discussed. PMID- 14565226 TI - An industrial process for synthesizing Lodenosine (FddA). AB - Two industrial synthetic approaches to Lodenosine (1, FddA, 9-(2,3-dideoxy-2 fluoro-beta-D-threo-pentofuranosyl) adenine) via a purine riboside or a purine 3' deoxyriboside are described. Several novel applications of deoxygenation and fluorination methods are compared considering reaction yields, economy, safety and environmental concerns. PMID- 14565227 TI - Ribonucleotide reductases: radical chemistry and inhibition at the active site. AB - Ribonucleoside 5'-diphosphate reductases (RDPRs) have been studied for several decades. Increasingly sophisticated mechanisms have been proposed for the reduction of natural substrate ribonucleotides to their 2'-deoxy counterparts and for mechanism-based inactivation of RDPRs with 2'-substituted-ribonucleotides. We now discuss biomimetic reactions of model substrate and inhibitor analogues, which clarify three aspects of previously proposed mechanisms postulated to occur at the active site of RDPRs. PMID- 14565229 TI - Recent advances in the synthesis of conformationally locked nucleosides and their success in probing the critical question of conformational preferences by their biological targets. AB - The present work describes some recent approaches to the syntheses of three classes of locked-North nucleosides: beta-D-ribo-, beta-D-deoxyribo-, and beta-D dideoxyribonucleosides. The method developed for the latter class permitted access to a novel bicyclo[3.1.0]hexene-type nucleosides structurally similar to D4T and carbovir. A structural analysis and biological activities are discussed. PMID- 14565228 TI - Inhibition of transcription by bisPNA-peptide conjugates. AB - Homopyrimidine bisPNAs have been reported to arrest transcription elongation by invading double-stranded DNA and forming a stable (PNA)2/DNA complex. We previously reported that attachment of a designed cationic peptide to the bisPNA enhances the efficiency of strand invasion. Here we investigate whether conjugation to cationic peptides can also improve inhibition of transcription. We observe that a conjugate between a bisPNA and a peptide containing eight lysines is a superior agent for inhibition of transcription, but that inhibition of transcription is reduced as pH and the concentration of magnesium are increased. Our studies provide useful characterization of bisPNAs as agents for inhibiting transcription. PMID- 14565230 TI - Structures of non-canonical tandem base pairs in RNA helices: review. AB - The structures of tandem non-canonical base pairs, a frequently recurring motif in RNA molecules, are reviewed and analysed. The tandem non-canonical base pair motifs can be roughly divided in three groups, containing seven subgroups based on their base pairing patterns and local geometries. Structural details and helical parameters that can be used to numerically distinguish between the subgroups are tabulated. Remarkably, while the individual helical twists of the tandem and adjacent base pair steps can be substantially smaller or larger than the typical A-form value of 32.7 degrees, the average value is close to A-form. This and other striking regularities resulting from compensating geometrical adjustments, important for understanding and predicting the configurations of non canonical base pairs geometries are discussed. PMID- 14565231 TI - Pyrazolo[3,4-d]pyrimidine nucleic acids: adjustment of the dA-dT to the dG-dC base pair stability. AB - The pyrazolo[3,4-d]pyrimidine-4,6-diamine nucleosides 2b-d stabilize the dA-dT base pair significantly when the dA-residue is replaced. Oligonucleotide duplexes incorporating 2b-d show a 4-6 degrees C Tm increase per modification. The 7-bromo compound 2b harmonizes the stability of the dA-dT vs. the dG-dC pair. According to this the stability of such duplexes depends no longer on the base pair composition of a DNA molecule. PMID- 14565232 TI - Oligonucleotide N3' --> P5' thio-phosphoramidate telomerase template antagonists as potential anticancer agents. AB - Human telomerase is a reverse transcriptase that is expressed in essentially all cancer cells, but not in the vast majority of normal somatic cells. Therefore, the specific inhibition of telomerase activity in tumors might have significant beneficial therapeutic effects. We have designed and evaluated oligonucleotide N3' --> P5' thio-phosphoramidates as telomerase template antagonists. In biochemical cell-free assays 11-13-mer thio-phosphoramidate oligonucleotides demonstrated sequence specific and dose dependent inhibition of telomerase with pico-molar IC50 values. Optimization of the oligonucleotide sequence and length resulted in the identification of a 13-mer-oligonucleotide thio-phosphoramidate GRN163 as a drug development candidate. In cell cultures GRN163 was able to inhibit telomerase activity in the absence of cationic lipid with approximately 1 microM IC50 values. Telomerase inhibition by GRN163 produced gradual telomere shortening, followed by cellular senescence and/or apoptosis of cancer derived cell lines. PMID- 14565233 TI - MDPSCL2: a new protecting group for chemoselective synthesis of 2'-O-alkylated guanosines. AB - An improved strategy for the synthesis of 2'-O-methyl-guanosine (6) and 2'-MOE guanosine (8) is reported. The regioselectivity of the alkylation was attained using a novel silicon-based protecting group, methylene-bis (diisopropyl silylchloride) (MDPSCl2, 2). The alkylation proceeded in a chemoselective manner using NaHMDS as the base and MeCl or MOE-Br as the appropriate electrophiles. PMID- 14565234 TI - Synthesis and biological evaluation of halo-neplanocin A as novel mechanism-based inhibitors of S-adenosylhomocysteine hydrolase. AB - Halogenated analogues of neplanocin A were synthesized from the key intermediate 1, among which fluoro-neplanocin A was found to be novel mechanism-based irreversible inhibitor of S-Adenosylhomocysteine hydrolase. PMID- 14565235 TI - Phosphono peptide nucleic acids with a constrained hydroxyproline-based backbone. AB - DNA mimics representing negatively charged analogues of peptide nucleic acids (PNAs), particularly hetero-oligomers constructed from alternating phosphono-PNA residues (pPNA) and monomers on the base of trans-4-hydroxy-L-proline (HypNA) as well as mimics composed of phosphono-HypNA monomers (pHypNA) were tested in a set of in vitro and in vivo assays, and they demonstrated a high potential for the use in nucleic acid based diagnostic, isolation of nucleic acids and antisense experiments. PMID- 14565236 TI - LNA and alpha-L-LNA: towards therapeutic applications. AB - LNA and alpha-L-LNA are promising candidates for the development of efficient oligonucleotide-based therapeutic agents. Here, we report dose-dependent inhibition of HIV-1 Tat-dependent trans activation by a 12-mer chimeric alpha-L LNA/DNA oligomer. This oligomer exhibits a dose-dependency similar to that of the corresponding 12-mer chimeric LNA/2'-O-Me-RNA oligomer. In addition, we show that incorporation of alpha-L-LNA or LNA monomers into each of the two binding arms of a "10-23" DNAzyme markedly increases cleavage of the target RNA. PMID- 14565237 TI - Synthesis and biological evaluation of beta-D-pentofuranonucleoside derivatives of 2-azidoadenine and 6-azidopurines. AB - Beta-D-pentofuranonucleoside derivatives of 2-azidoadenine and 6-azidopurines have been synthesized. The azido-tetrazolo tautomerism observed on such nucleoside analogues has been studied. The compounds were tested for their activity against HIV and HBV but they did not show significant antiviral effect. PMID- 14565238 TI - 2'-Fluoro-4'-thio-2',3'-unsaturated nucleosides: anti-HIV activity, resistance profile, and molecular modeling studies. AB - Both D- and L-2'-fluoro-4'-thio-2',3'-unsaturated nucleosides were synthesized and their anti-HIV activity against the drug sensitive virus and lamivudine resistant mutant (M184V) were evaluated. In vitro antiviral evaluation indicated that the L-isomers are more potent than the D-isomers, but unfortunately all were cross-resistant with 3TC. Molecular modeling studies revealed that the unnatural sugar moiety of the L-nucleosides as well as 4'-sulfur atom of the D-isomer has a steric conflict with the bulky side chain of valine 184, resulting in cross resistance. PMID- 14565239 TI - Developing synthetic methods for bioactive phosphorus compounds using H phosphonate chemistry: a progress report. AB - In this paper a short account of our recent research concerning the development of new synthetic methods and reagents for the preparation of nucleotides and their analogues, is given. PMID- 14565240 TI - Preparation and properties of a new type of acyclic, achiral nucleoside analogue. AB - Preparation of the nucleoside analogues 1 and incorporation of 1, B = T, in deoxyribooligonucleotides by the phosphoramidite method is described. A two-step deprotection procedure was developed to reduce cleavage of the modified allylic unit. The binding properties of the modified oligonucleotides towards complementary DNA and RNA has been evaluated by Tm measurements showing a deltaTm of -2 to -6.5 degrees C per modification. An oligonucleotide with two modifications at the 3'-end showed considerable resistance towards cleavage by a 3'-exonuclease. No antiviral activity against HIV-1 or HSV-1 was found for 1, B = G or T, or for any of the trihydroxy derivatives 5. PMID- 14565241 TI - A short, novel, and cheaper procedure for oligonucleotide synthesis using automated solid phase synthesizer. AB - Dimethylthiuram disulfide (DTD) has been developed as an efficient thiolation reagent during automated synthesis of oligonucleotides using phosphoramidite chemistry. Simultaneous thiolation and capping was accomplished by mixing DTD with capping solution B, which saved 20% of solvent consumption and compressed the four-step synthesis cycle to three. Large-scale (1 mmol) synthesis of phosphorothioate oligonucleotides has been demonstrated with excellent yield and purity. PMID- 14565242 TI - Convenient synthesis of oligodeoxynucleotides containing 2'-deoxy-6-thioinosine. AB - A facile synthesis of oligodeoxynucleotides (ODN) containing 2'-deoxy-6 thioinosine (dI6S) based on the convertible nucleoside O6-phenyl-2'-deoxyinosine is presented. After standard solid-phase DNA synthesis and removal of the cyanoethyl protecting groups with DBU treatment with aqueous sodium hydrogen sulfide introduces the sulfur functionality, deprotects the other nucleobases and cleaves the ODN from the solid support in a one-pot reaction. In addition, the extinction coefficient of 2'-deoxy-6-thioinosine is determined by enzymatic fragmentation of the resulting ODN in the presence of adenosine deaminase. PMID- 14565243 TI - A role for oligonucleotide-based RNA-knock down technologies in functional genomics. AB - Functional genomics is inundating the pharmaceutical industry with large numbers of potential gene targets from several sources such as gene expression profiling experiments (DNA microchips, proteomics) or database mining. Oligonucleotide based RNA-knock down technologies such as antisense or RNA interference can aid in the filtering and prioritization of target candidates in the drug discovery process. PMID- 14565244 TI - Properties of triple helices formed by oligonucleotides containing 8 aminopurines. AB - The synthesis of parallel hairpins carrying 8-aminopurines is described. These hairpins have a high affinity for specific polypyrimidine sequences resulting in the formation of very stable triplexes. PMID- 14565245 TI - Labeling during cleavage of nucleic acids for their detection on DNA chips. AB - A new and efficient strategy for labeling of nucleic acids prior to their hybridization on high density DNA chip has been developed. Our approach which combines the fragmentation and the labeling is based on the reactivity of the terminal phosphates of cleaved DNA and RNA fragments with a reporter molecule bearing aryldiazomethane group. PMID- 14565246 TI - Structural basis for topoisomerase I inhibition by nucleoside analogs. AB - Nucleoside analogs such as 1-beta-D-arabinofuranosyl cytidine (AraC) and 2',2' difluoro deoxycytidine (dFdC) are important components of the anticancer chemotherapeutic arsenal and are among the most effective anticancer drugs currently available. Although both AraCTP and dFdCTP impede DNA replication through pausing of DNA polymerases, both nucleoside analogs are ultimately incorporated into replicated DNA and interfere in DNA-mediated processes. Our laboratories are investigating the structural basis for the poisoning of topoisomerase I (top1) due to antipyrimidine incorporation into duplex DNA. We recently reported that both AraC and dFdC induce formation of top1 cleavage complexes, and poisoning of top1 contributes to the anticancer activities of both these drugs. Recent NMR and thermodynamic studies from our laboratories provide insight into the mechanism by which AraC and dFdC poison top1. NMR studies from our laboratories have revealed that the arabinosyl sugar of AraC adopted a C2' endo conformation. Although this is a B-type sugar pucker characteristic of duplex DNA, the conformation is rigid, and this lack of flexibility probably contributes to inhibition of the religation step of the top1 reaction. In contrast to AraC, NMR studies revealed dFdC adopted a C3' endo sugar pucker characteristic of RNA, rather than DNA duplexes. dFdC substitution enhanced formation of top1 cleavage complexes, but did not inhibit religation. The enhancement of top1 cleavage complexes most likely results from a combination of conformational and electrostatic effects. The structural effects of dFdC and AraC are being further investigated in duplex DNA with well-defined top1 cleavage sites to analyze more specifically how these structural perturbations lead to enzyme poisoning. PMID- 14565247 TI - Synthesis of (Z)-(1-fluoro-2-hydroxymethylcyclopropylmethyl)purines. AB - (Z)-(1-fluoro-2-hydroxymethylcyclopropylmethyl)purines were designed, synthesized and evaluated their antiviral activity against poliovirus, HSV, and HIV. PMID- 14565248 TI - Cyclic uridine diphosphate glucose: a new pyrimidine analog of cyclic ADP ribose. AB - Novel compound 1, as the first example of cyclic ADP-ribose analogs containing a pyrimidine residue, was synthesized by a chemical strategy employing a Mitsunobu reaction for the condensation of the glucosyl moiety on protected uridine, and a Matsuda procedure for the cyclization step. PMID- 14565249 TI - A new approach for syntheses of 2',3'-dideoxy-2',3'-dehydronucleosides using 2,2 difluoro-1,3-dimethylimidazolidine (DFI) as a dehydrating reagent. AB - We found that 2,2-difluoro-1,3-dimethylimidazolidine (DFI) is useful for not only fluorination but also dehydrating reactions. This dehydrating ability of DFI was applied to the syntheses of dihydrofurans (2) that are possible starting materials for various anticancer or antiviral drugs. PMID- 14565250 TI - Synthesis of new pseudonucleosides containing chiral cyclosulfamides as agycone. AB - A series of chiral cyclosulfamides have been synthesized in four steps, starting from N-benzoylaminoacids. Regioselective glycosylation of these pseudopyrimidic heterocycles was carried out after deprotection. Best glycosylation results were obtained by preliminary silylation of cyclosulfamides, and their condensation with a tetraacetylribofuranose and pentaacetylglucopyranose is described, which yielded the pseudonucleosides in a beta-anomeric configuration. PMID- 14565251 TI - Synthesis of thymine derivatives of 4-hydroxyvaline. AB - A synthetic route to thymine derivatives of (2S,3R)- and (2S,3S)-4-hydroxyvaline has been developed starting from commercially available L-aspartic acid. PMID- 14565252 TI - Doped natural phosphate: a new and environmentally friendly catalyst in nucleoside synthesis. AB - Doped natural phosphate is used as acidic or basic catalyst in nucleoside and acyclonucleoside synthesis. Some examples are given. PMID- 14565253 TI - Synthesis of carbocyclic analogues of thymidine. AB - A new route towards an enantiomerically pure carbocyclic 2'-deoxyribonucleoside precursor was developed. After coupling with a nucleobase the product is easily accessible for further modifications at the 3'-hydroxy group. PMID- 14565254 TI - The regiospecific one-pot phosphorylation of either the 5'- or 2'-hydroxyl in 3' deoxycytidines without protection: critical role of the base. AB - We report methodology which enables direct phosphorylation of 3'-deoxycytidine exclusively either at the 5'-hydroxyl or the 2'-hydroxyl. Protection of the base is not required. Standard phosphoramidochloridates in combination with pyridine and tert-butyl magnesium chloride is employed, in which the ratio of nucleoside to Grignard reagent is crucial. These findings, which appear to be general for 3' deoxycytidines, are not applicable to 3'-deoxyuridine or 3'-deoxyguanosine. PMID- 14565256 TI - New solid supports linking nucleoside scaffolds. AB - An easy and efficient strategy to obtain new nucleoside based solid supports in which the nucleoside moieties have been anchored to the solid support through the nucleobase is here proposed. A simple and efficient solid-phase synthesis of 5' and 3'-derivatized uridine analogues has so been developed, following methodologies well established in organic chemistry. PMID- 14565255 TI - Synthesis of novel mRNA 5' cap-analogues: dinucleoside P1, P3-tri-, P1, P4-tetra , and P1, P5-pentaphosphates. AB - A series of new mRNA anti reverse cap analogues (ARCA) was designed to obtain a tool for studying the mechanism of protein translation. Dinucleoside P1, P3-tri-, P1, P4-tetra- and P1, P5-pentaphosphates, linked by a 5'-to-5' phosphate bridge and composed of modified 7-methylguanosine and guanosine, have been synthesized. The hydroxyl group (2'OH or 3'OH) in 7-metylguanosine moiety was replaced by OCH3 or -H in order to obtain the cap analogues capable to be correctly incorporated into synthetic mRNA transcripts. Tri-, tetra-, and pentaphosphates were prepared by ZnCl2 catalyzed condensation in DMF of derivatives of the 7 methylguanosine diphosphates with the guanosine mono-, di- and triphosphate P imidazolides, respectively. The structures of the novel compounds were established by means of 1H and 31P NMR spectra. PMID- 14565257 TI - One-pot synthesis of an AZT boranophosphate conjugated with tyrosine: a potential prodrug candidate. AB - A one-pot synthesis of P-tyrosinyl(P-O)-5'-P-AZT boranophosphate 7 via a phosphoramidite method is described. The P-boranophosphate diastereomers were separated by RP-HPLC, and their structures were confirmed by NMR and MS. PMID- 14565258 TI - Development of a nucleoside analog UV light sensor. AB - Conjugation of the photosensitive nucleoside (E)-5-(2 methoxycarbonylethenyl)cytidine to biotin provided a means to attach this analogue to microparticles for dosimetry applications that require UV sensor mobility. PMID- 14565259 TI - 4'-C-methyl-beta-D-ribofuranosyl purine and pyrimidine nucleosides revisited. AB - In order to evaluate their antiviral properties, a series of 4'-C-methyl-beta-D ribofuranosyl purine and pyrimidine nucleosides has been prepared. Unfortunately, none of these 4'-branched nucleosides showed any antiviral activity or cytotoxcity when tested against HIV, HBV, and Yellow Fever virus. PMID- 14565260 TI - A facile synthetic method for 3'-alpha-fluoro-2',3'-dideoxyadenosine. AB - A facile method for the synthesis of 3'-alpha-fluoro-2',3'-dideoxyadenosine (5) has been developed using a novel rearrangement of 3'-beta-bromine to the 2'-beta position during 3'-alpha fluorination. PMID- 14565261 TI - A zero-length diazirine photoactive nucleoside. AB - The scheme of synthesis which allows to obtain 5-(3H-diazirin-3-yl)-2' deoxyuridine as the zero-length photoactive nucleoside is described. PMID- 14565262 TI - Synthesis and biological evaluation of pyrimidine nucleosides fused with 3',4' tetrahydrofuran ring. AB - Pyrimidine nucleosides fused with 3',4'-tetrahydrofuran ring were synthesized, starting from 1,2;5,6-di-O-isopropylidene-D-glucose and assayed for antiviral activities. Thymine analogue 1 and its corresponding 2'-deoxy analogue 3 exhibited high cytotoxicity instead of giving antiviral activities. PMID- 14565264 TI - Synthetic studies on guanofosfocin: glycosylation of 8-oxo-purine nucleosides via Mitsunobu reaction. AB - In model studies directed to the total synthesis of guanofosfocins, a unique glycosidic bond formation between the 8-oxo-purine nucleosides and mannopyranose derivatives under Mistunobu conditions is described. PMID- 14565263 TI - Bi- and tricyclic nucleoside derivatives restricted in S-type conformations and obtained by RCM-reactions. AB - Ring-closing metathesis (RCM) is applied as a new and powerful technology in the construction of nucleoside analogues that are conformationally restricted in S type conformations due to additional 3',4'- and/or 3',5'-linkages. PMID- 14565265 TI - A new type of acyclic, achiral nucleoside analogue. How does it simulate nucleosides? AB - The new monomer 1 seems to be an excellent mimic of nucleosides with different sugar conformations (north, south, and envelope), because of the relatively free rotation around gamma, delta, and chi. The rotation around chi is primarily controlled by the repulsion between H6 and the two hydrogen atoms on C4' and not pi conjugation between the double bond and the nucleobase. A viable synthesis of the guanine monomer 8 is described. PMID- 14565266 TI - Rearrangement reactions of guanosine cyclonucleosides and their analogs. AB - Under acid-catalyzed transglycosylation conditions 5',8-cyclo-8-oxoguanine nucleosides undergo a ring-opening reaction to 8-oxoguanine derivatives, instead of the 7-9 isomerization. PMID- 14565267 TI - Diastereo- and enantioselective synthesis of 1'-C-branched N,O-nucleosides. AB - A synthetic approach towards 1'-C-branched N,O-nucleosides is reported, based on 1,3-dipolar cycloaddition of ethoxycarbonylnitrone. The asymmetric version of the process exploits the presence of a chiral auxiliary at the carbon atom of nitrone and leads to beta-D and beta-L nucleosides in good yields. PMID- 14565268 TI - Reactivity models of 1-N-vinyluracil and synthesis of a new class of potential antiviral agents by the use of 1,3-dipolar cycloaddition reactions. AB - By the use of a convergent approach based on 1,3-dipolar cycloaddition reactions between N-protected formylnitrones generated in situ and 1-N-vinyluracil, a new class of 4'-aza-analogues of 2',3'-dideoxynucleosides is synthesized. Competitive reaction for the endocyclic bond of uracil also brings to a new isoxazolidine derivative fused with the pyrimidine nucleus. PMID- 14565269 TI - Selective metalation of 6-methylpurines: synthesis of 6-fluoromethylpurines and related nucleosides. AB - A selective metalation at the 6-CH3 over C-8 of 6-methylpurine derivative 6 was observed with softer counter cation (Na+ or K+) of the base, while the harder Li+ showed no selectivity. In the presence of N-fluorobenzenesulfonamide (NFSI), this property was utilized for the synthesis of 6-fluoromethylpurine derivatives 4 and 5 as potential toxins for suicide gene therapy. PMID- 14565270 TI - Chemo-enzymatic synthesis of 3-deoxy-beta-D-ribofuranosyl purines and study of their biological properties. AB - 9-(3-Deoxy-beta-D-erythro-pentofuranosyl)-2,6-diaminopurine (2) was synthesized by an enzymatic transglycosylation of 2,6-diaminopurine using 3'-deoxycytidine (1) as a donor of the sugar moiety. Nucleoside 2 was transformed to 3'-deoxy guanosine (3), 9-(3-deoxy-beta-D-erythro-pentofuranosyl)-2-amino-6-oxopurine (3' deoxyisoguanosine; 4), and 9-(3-deoxy-beta-D-erythro-pentofuranosyl)-2 fluoroadenine (5). Compounds 2-5 were evaluated for their anti-HIV activity. PMID- 14565271 TI - Efficient synthesis of 8-thiosubstituted guanine derivatives as potential tools for biochemical and biological studies. AB - A method for the selective introduction of the N2-(dimethylamino)methylene group into 8-thio-9-(2-hydroxyethoxymethyl)guanine (1) has been developed. The effect of the N2-amidine protection on the S-alkylation of 1 was studied. PMID- 14565272 TI - Synthesis of carbocyclic analogues of MECA and NECA 1,2-disubstituted as potential adenosine receptor agonists. AB - A new class of 1,2-disubstituted carbocyclic nucleosides of MECA and NECA analogues was synthesized in good yield starting from (+/-) 6 azabicyclo[3.2.0]heptan-7-one. PMID- 14565273 TI - 4-Thiofuranoid glycals: versatile synthons for stereoselective synthesis of 4' thionucleosides. AB - Beta-anomers of 4'-thionucleosides have been synthesized stereoselectively, through PhSeCl- or N-iodosuccimide (NIS)-initiated electrophilic glycosidation to 3,5-O-(di-t-butylsilylene)-4-thiofuranoid glycal (1). This synthetic method has been applied to the synthesis of those analogues branched at the anomeric position using 1-C-carbon-substituted 3,5-O-(tetraisopropyldisiloxane-1,3-diyl)-4 thiofuranoid glycals (11-14) prepared based on lithiation of 10. PMID- 14565274 TI - Stannylation approach to the synthesis of 2'- and 3'-substituted analogues of 2',3'-didehydro-2',3'-dideoxynucleosides. AB - Three methods are described for the introduction of a tributylstannyl group to the sp2-carbon of 2',3'-didehydro-2',3'-dideoxy nucleosides (d44Ns). The resulting stannylated products serve as versatile intermediates for the synthesis of d4Ns having various types of carbon-substituent. PMID- 14565275 TI - Practical synthesis of D-cyclopent-2-enone, the key intermediate of carbocyclic nucleosides. AB - An efficient and practical method for the synthesis of (4R,5R)-4,5-O isopropylidene-cyclopent-2-enone was developed from D-ribose by using a ring closing metathesis reaction. PMID- 14565276 TI - Covalent analogues of nucleobase-pairs. AB - Covalently linked base pairs analogues consisting of purine-purine or purine pyrimidine conjugates linked by carbon linkages of diverse length and configuration (ethylene, vinylene, acetylene and phenylene) were prepared. PMID- 14565277 TI - Introduction of a benzyl group onto the 2'-OH of 6-chloropurine 3'-O benzoylriboside. AB - A new method to introduce a benzyl group onto the 2'-OH of purine ribonucleoside is described. Thus, 6-chloropurine 3'-O-benzoylriboside and its 5'-O-trityl congener were condensed with benzyl alcohol using the Mitsunobu reaction to give the 2'-O-benzyl derivative. The yields were varied from 4.6 to 62.9% depending on the solvent. The product was converted to adenosine, indicating that the stereochemistry at C-2' is retained. PMID- 14565278 TI - Stereocontrolled synthesis of diene and enyne sugar-modified nucleosides and their interaction with S-adenosyl-L-homocysteine hydrolase. AB - Conjugated diene 5-7 and enyne 8 analogs derived from adenosine and uridine were synthesized employing Pd-catalyzed cross-coupling reactions. PMID- 14565279 TI - Purine derivatives of 1,2-disubstituted cyclohexane analogues of nucleosides. AB - Starting from (+/-)-cis-2-hydroxymethylcyclohexylamine, a series of cyclohexane derived cis-1,2-disubstituted carbonucleoside analogues with a 6- or 2,6-purine or 8-azapurine base were synthesized. The antiviral and antitumoral in vitro effects of the new compounds were evaluated. PMID- 14565280 TI - Study of different substituted cyclic and acyclic benzylpronucleotides of d4T relative to their hydrolytic stability and antiviral activity. AB - CycloSal-d4TMP and two different bis(benzyl) phosphate triesters of the antivirally active nucleoside analog d4T were studied with regard to their chemical hydrolysis behavior at pH 7.3, in CEM/0 cell extracts, and their anti HIV activity. In contrast to triesters 2-4, bis-(o-AB)-d4TMP 1 was found to be chemically exquisitely stable. All compounds led to the formation of d4TMP in cell extracts and all triesters achieved the TK-bypass. PMID- 14565281 TI - Stereochemical analysis of diastereomeric 1,3-bis(adenosine-5'-O phosphorothioyl)glycerols. PMID- 14565282 TI - Design of Mycobacterium tuberculosis thymidine monophosphate kinase inhibitors. PMID- 14565283 TI - 5'-O-ester prodrugs of potent and selective anti-HIV agent--2',3'-dideoxy-3' fluoro-2-thiothymidine (S2FLT): synthesis and anti-HIV activity. AB - Novel synthesis of 2',3'-dideoxy-3'-fluoro-2-thiothymidine (SFLT) based on transformation of appropriately protected 1-beta-D-threo-ribofuranosylthymine is presented. The synthesis and evaluation of SFLT 5'-O-ester prodrugs enzymatic hydrolysis, as well as their anti-HIV activity, is also described. PMID- 14565284 TI - 2-Phenylhydroxypropynyladenosine derivatives as high potent agonists at A2B adenosine receptor subtype. AB - Adenosine derivatives bearing in 2-position the (R,S)-phenylhydroxypropynyl chain were evaluated for their potency at human A2B adenosine receptor, stably transfected on CHO cells, on the basis that (R,S)-2-phenylhydroxy-propynyl-5'-N ethylcarboxyamidoadenosine [(R,S)-PHPNECA] was found to be a good agonist at the A2B receptor subtype. Biological studies demonstrated that the presence of small alkyl groups in N6-position of these molecules are well tolerated, whereas large groups abolished A2B potency. On the other hand, the presence of an ethyl group in the 4'-carboxamido function seems to be optimal, the (S)-PHPNECA resulting the most potent agonist at A2B receptor reported so far. PMID- 14565285 TI - Synthesis and biological activity of 2-aminopurine methylenecyclopropane analogues of nucleosides. AB - Synthesis and biological activity of 7- and 9-isomers (Z+E) of methylenecyclopropane analogues of 2-aminopurine nucleosides is described. The (S,Z)-9-isomer is a substrate for xanthine oxidase. PMID- 14565286 TI - Synthesis of unusual bicyclic nucleosides bearing an unsaturated side-chain, as potential inhibitors of varicella-zoster virus (VZV). PMID- 14565287 TI - Novel nucleotide analogues as potential substrates for TMPK, a key enzyme in the metabolism of AZT. AB - Novel cyclic and acyclic analogues of dTMP and AZTMP were synthesized from the corresponding cycloSal-phosphotriesters. This method yielded the nucleotides in good yields with a simple work-up. Investigation of the substrate properties of the modified nucleotides towards TmpK showed, that they are very poor substrates for this key enzyme in the bioactivation of AZT. PMID- 14565288 TI - d4TMP delivery from 7-substituted cycloSal-d4TMPs. AB - Benzyl-substituted cycloSal-d4T monophosphates were prepared and evaluated for their ability to release d4TMP selectively. In contrast to previously reported derivatives, two of the new compounds release d4TMP as the sole product while two others gave the expected benzyl phosphate diesters. However, these diesters were surprisingly stable against degradation to release d4TMP. PMID- 14565289 TI - Synthesis of new homo and heterodimers of 2',3'-dideoxyinosine (ddI) using ester linkages. AB - A series of new homo and heterodimers of ddI has been synthesized. A glutarate diester spacer was used to covalently couple ddI onto ddI, AZT or d4T. PMID- 14565290 TI - Synthesis and antiviral activity assay of novel (E)-3',5'-diamino-5-(2 bromovinyl)-2',3',5'-trideoxyuridine. AB - (E)-3',5'-diamino-5-(2-bromovinyl)-2',3',5'-trideoxyuridine (5), the diamino analogue of BVDU (1), was synthesized from BVDU. In contrast with BVDU, compound 5 did not show activity against herpes simplex virus or varicella-zoster virus. PMID- 14565292 TI - Non-inhibition of acetylcholinesterase by cycloSal nucleotides. AB - An acetylcholinesterase (AChE) assay based on the Rappaport method was established to investigate the behaviour of several cycloSal nucleotides against AChE from electrophorus electricus and human sources (purified enzymes). AChE is a physiologically essential enzyme as it catalyzes the hydrolysis of the neurotransmitter acetylcholine. No inhibition was observed in any of the cases. PMID- 14565291 TI - Optimization of antiviral prodrug properties using combinatorial methods. AB - Some diacid biodegradable synthesis of aziduthymidine (AZT) were synthesized and applied to production of about 60 different derivatives. PMID- 14565293 TI - Synthesis and biological evaluation of a series of new cyclohexenyl nucleosides. AB - A series of cyclohexenyl nucleosides (1-8) were successfully prepared with moderate yield and their cytotoxicity and antiviral activity were investigated. Among the eight new compounds, only the diaminopurine analogue 8 showed pronounced activity against HSV-1, HSV-2. PMID- 14565294 TI - Synthesis and antiviral evaluation of ribavirin congeners containing a hexitol moiety. AB - Several ribavirin congeners containing a hexitol moiety were prepared via ring opening of two different epoxides with the methylcarboxylate ester of triazole and further elaboration. Unfortunately, none of the newly synthesized compounds displayed appreciable antiviral activity. PMID- 14565295 TI - Dinucleoside monophosphates containing AZT and 1-methyladenosine or 7 methylguanosine. AB - Dinucleoside monophosphates containing AZT and 1-methyladenosine or 7 methylguanosine were synthesized and their in vitro anti-HIV activity was determined. PMID- 14565296 TI - 2'-Deoxy-2'-C-trifluoromethyl beta-D-ribonucleoside analogues: synthesis and antiviral evaluations. AB - Hitherto unknown 2'-deoxy-2'-C-trifluoromethyl-beta-D-ribonucleoside derivatives bearing the five naturally occurring nucleic acid bases have been synthesized. The compounds were tested for their activity against HIV, HBV and several RNA viruses, but they did not show significant antiviral effect. PMID- 14565297 TI - Synthesis and antiviral evaluation of beta-D- and beta-L-pentofuranonucleoside derivatives bearing 5-trifluoromethylcytosine as the base. AB - Beta-D- and beta-L-pentofuranonucleoside derivatives bearing 5 trifluoromethylcytosine as the base have been synthesized. The compounds were tested for their activity against HIV and HBV, but they did not show significant antiviral effect. PMID- 14565298 TI - Dinucleoside polyphosphate NAD analogs as potential NMN adenylyltransferase inhibitors. Synthesis and biological evaluation. AB - Two dinucleoside polyphosphate NAD analogs, P1-(adenosine-5')-P3-(nicotinamide riboside-5')triphosphate (Np3A, 1) and P1-(adenosine-5')-P4-(nicotinamide riboside-5')tetraphosphate (Np4A, 2), were synthesized and tested as inhibitors of both microbial and human recombinant NMN adenylyltransferase. Compounds 1 and 2 proved to be selective inhibitors of microbial enzymes. PMID- 14565299 TI - A new tiazofurin pronucleotide: synthesis and biological evaluation of cyclosaligenyl-tiazofurin monophosphate. AB - Synthesis and biological activities of cyclosaligenyl-tiazofurin monophosphate (CycloSal-TRMP), a new tiazofurin pronucleotide, are reported. CycloSal-TRMP proved to be active in vitro against human myelogenous leukemia K562 cell line and as A1 adenosine receptor agonist. PMID- 14565300 TI - Synthesis of certain heterodimers expected as HIV-1 reverse transcriptase inhibitors. AB - Expected for the ability to inhibit HIV replication, we report the synthesis of two heterodimers of the general formula: [2NRTI]-C5-GLY-SUCCINYL-Npiperazinyl [NNRTI] (18, 19) containing both a Nucleoside Reverse Transcriptase Inhibitor (10, 11) and a Non-Nucleoside Reverse Transcriptase Inhibitor (8) [Trovirdine Analogue belonging of the phenethyl thiazolyl thiourea class] connected through the "succinyl-glycine" spontaneously cleavable linker. PMID- 14565302 TI - Thionucleotides as inhibitors of ribonucleotide reductase. AB - Ribonucleosides and xylonucleosides bearing a disulfide function on the sugar ring were synthesized. Ribonucleosides belonging to the cytidine series were found to efficiently reduce dNTP pools in the human lymphoblastoid CEM/SS cell line. PMID- 14565301 TI - Deaza- and deoxyadenosine derivatives: synthesis and inhibition of animal viruses as human infection models. AB - N6-Cycloalkyl-2',3'-dideoxyadenosine derivatives and (2-chloro)-N6-cycloheptyl-3 deazaadenosine have been synthesized and tested, along with other (deaza)purine (deoxy)nucleosides from our chemical library, as inhibitors of virus replication against Bovine Herpes Virus 1 (BHV-1) and sheep Maedi/Visna Virus (MVV). Most compounds demonstrated good antireplicative activity against MVV, showing also low cell toxicity. PMID- 14565303 TI - Synthesis of 4'-C-ethynyl and 4'-C-cyano purine nucleosides from natural nucleosides and their anti-HIV activity. AB - Purine 2'-deoxynucleosides bearing an ethynyl or a cyano group at C-4' of the sugar moiety were synthesized from the corresponding 2'-deoxynucleosides. These compounds exhibited very potent anti-HIV activity, and remained active against drug resistant HIV strains. PMID- 14565304 TI - Synthesis and antiviral evaluation of 2',3'-dideoxy-2'-fluoro-3'- C-hydroxymethyl beta-D-arabinofuranosyl pyrimidine nucleosides. AB - The synthesis and anti-HBV and anti-HIV activity of a number of 2',3'-dideoxy-2' fluoro-3'-C-hydroxymethyl-beta-D-arabinofuranosyl pyrimidine nucleosides are reported. PMID- 14565305 TI - Investigations of nucleoside H-phosphonamidate in the design of nucleotide prodrug. AB - The synthesis, anti-HIV activity and stability studies of a H-phosphonamidate derivative of 3'-azido-2',3'-dideoxythymidine (AZT) incorporating a N,N diisopropylamino residue as first model of alkylamino group are reported. The results demonstrate that such phosphorylated structure exerts its biological effects via chemical hydrolysis into the corresponding H-phosphonate, precursor of the parent nucleoside. PMID- 14565306 TI - Mononucleoside SATE glucosyl phosphorothiolates as a new series of pronucleotides. AB - The synthesis and the study of two phosphorothiolate derivatives of 3'-azido 2',3'-dideoxythymidine (AZT) bearing a S-pivaloyl-2-thioethyl (tBuSATE) group and glucosyl residues associated to the phosphorus atom by a 2-oxyethyl link, are reported. These derivatives could be considered as prototypes of a new series of nucleotide prodrugs (pronucleotides). PMID- 14565307 TI - SATE (aryl) phosphotriester series. I. Synthesis and biological evaluation. AB - Synthesis and biological activities of several phosphotriester derivatives of 3' azido-2',3'-dideoxythymidine (AZT) bearing a S-pivaloyl-2-thioethyl (tBuSATE) group and aryl residues derived from L-tyrosine are reported. All compounds showed marked anti-HIV activity in thymidine kinase-deficient CEM cells demonstrating their ability to deliver intracellularly the parent 5' mononucleotide. PMID- 14565308 TI - SATE (aryl) phosphotriester series. II. Stability studies and physicochemical parameters. AB - The stability of phosphotriester derivatives of 3'-azido-2',3'-dideoxythymidine (AZT) bearing a S-pivaloyl-2-thioethyl (tBuSATE) group and various aryl residues derived from L-tyrosine was evaluated in biological media. The results demonstrate that such compounds give rise to intracellular delivery of the parent mononucleotide through esterase and phosphodiesterase hydrolytic steps, successively. PMID- 14565309 TI - Synthesis and fluorescent properties of 6-(4-biphenylyl)-3,9-dihydro-9-oxo-5H imidazo[1,2-a]purine analogues of acyclovir and ganciclovir. AB - Tricyclic (T) analogues of acyclovir (ACV, 1) and ganciclovir (GCV, 2) carrying the 3,9-dihydro-9-oxo-5H-imidazo[1,2-a]purine system [i.e., 6-(4-BrPh)TACV, 5 and 6-(4-BrPh)TGCV, 6] were transformed into 6-[(4'-R2)-4-biphenylyl] derivatives of TACV (7-9) and TGCV (10-12) by Suzuki cross coupling with 4-substituted phenylboronic acids. Compound 11 (R2 = CH2OH) showed a high (approximately 1000) selectivity index against herpes simplex virus type 1 (HSV-1) together with advantageous fluorescence properties (emission in visible region, little overlap with absorption and moderate intensity). PMID- 14565310 TI - Synthesis of 5-azacytidine nucleosides with rigid sugar moiety as potential antitumor agents. AB - The bicyclic 3'-O,5'-C-methylene-linked and 2'-O,5'-C-methylene-linked 5 azacytidine derivatives were readily synthesized from 1,2;5,6-di-O-isopropylidene D-glucose and evaluated against several cancer cell lines. PMID- 14565312 TI - Design and synthesis of A3 adenosine receptor ligands, 3'-fluoro analogues of Cl IB-MECA. AB - Synthesis of 3'-deoxy-3'-fluoro-N6-substituted adenosines as bioisosteres of Cl IB-MECA and their binding affinities to A3 adenosine receptor are described. PMID- 14565311 TI - Synthesis of halogenated 9-(dihydroxycyclopent-4'-enyl) adenines and their inhibitory activities against S-adenosylhomocysteine hydrolase. AB - Novel halovinyl analogues of neplanocin A without 4'-hydroxymethyl group were easily synthesized starting from D-ribose via cyclopentenone 5 as a key intermediate and their inhibitory activity against SAH hydrolase was assayed. PMID- 14565313 TI - Design and synthesis of A3 adenosine receptor ligands, 2'-fluoro analogues of Cl IB-MECA. AB - Synthesis of 2'-deoxy-2'-fluoro-N6-substituted adenosines as bioisosteres of Cl IB-MECA and their binding affinities to A3 adenosine receptor are described. PMID- 14565314 TI - Bicyclic nucleoside inhibitors of varicella-zoster virus: 5'-chloro and 3'-chloro derivatives. AB - We have recently discovered bicyclic furopyrimidines as potent and selective inhibitors of VZV. In order to investigate the structural requirements for antiviral activity we have succesfully synthesised some 3'-chloro and 5'-chloro derivatives. The compounds have been tested against VZV and CMV, but displayed no significant in vitro activity. PMID- 14565315 TI - Bicyclic nucleoside inhibitors of varicella-zoster virus: synthesis and biological evaluation of 2',3'-dideoxy-3'-fluoro and 2'-deoxy-xylo derivatives. PMID- 14565316 TI - Synthesis of AZA analogues of TSAO. AB - TSAO derivatives which were first synthesized in 1992 have shown strong inhibitory effect and selectivity against HIV-1 (Camarasa, M.J.; Perez-Perez, M.J.; San-Felix, A.; Balzarini, J.; De Clercq, E. J. Med. Chem. 1992, 35, 2721 2727). The structure-activity relationship of these derivatives has shown strong binding between the amino acids constituting the reverse transcriptase and the different pharmacophore (tert-butyldimethylsilyl group, amino and sulfonate groups of the TSAO derivatives) (Camarasa, M.J.; San-Felix, A.; Perez-Perez, M.J.; Velazquez, S., Alvarez, R.; Chamorro, C.; Jimeno, M.L.; Perez, C.; Gago, F.; De Clercq, E.; Balzarini, J. J. Carbohydr. Chem. 2000, 19, 6403-6406). We described the synthesis of an original TSAO analogue where, basically, the O-1'' atom is replaced by a nitrogen atom. PMID- 14565317 TI - Synthesis, stability, and biological evaluation of 1,3-dihydrobenzo[c]furan analogue of d4T and its SATE pronucleotide. AB - The anti-HIV activity and stability studies of 1,3-dihydrobenzo[c]furan analogue of d4T are reported. The corresponding mononucleoside phosphotriester derivative bearing a S-pivaloyl-2-thioethyl (tBuSATE) group, as biolabile phosphate protection, is also studied. PMID- 14565318 TI - N-3 substituted TSAO derivatives as a probe to explore the dimeric interface of HIV-1 reverse transcriptase. PMID- 14565319 TI - Towards new thymidine phosphorylase/PD-ECGF inhibitors based on the transition state of the enzyme reaction. AB - Computational studies have been conducted to built a closed form of TPase and to characterize the transition state of the phosphorylisis reaction catalyzed by TPase. The results obtained point to a crucial role of His-85 and the O2 of thymine in the catalysis. This modelled transition state forms the basis for the design of new TPase inhibitors. PMID- 14565320 TI - Synthesis of [1'-fluoro-2',2'-bis-(hydroxymethyl)cyclopropylmethyl]purines as antiviral agents. AB - [1'-Fluoro-2',2'-bis-(hydroxymethyl)cyclopropylmethyl]purines were designed, synthesized and their antiviral activity against poliovirus, HSV and HIV was evaluated. PMID- 14565321 TI - Unusual lability of 5'-O-tert-butyldimethylsilyl group on 4''-acyl TSAO derivatives. PMID- 14565322 TI - Phosphorodiamides as prodrugs for antiviral nucleosides. AB - New phosphorodiamides of modified nucleoside monophosphates were synthesized and their antiviral properties were evaluated. PMID- 14565323 TI - Synthesis and biological evaluation of some acyclic 4,6-disubstituted 1H pyrazolo[3,4-d]pyrimidine nucleosides. AB - The chemical synthesis and biological evaluation of some acyclic alpha-[6-(1' carbamoylalkylthio)-1H-pyrazolo[3,4-d]pyrimidin-4-yl]thioalkylamide nucleosides are described. PMID- 14565324 TI - Thiated analogues of 2',3'-dideoxy-3'-fluorothymidine and their phosphorylated and phosphonylated derivatives: synthesis, interaction with HIV reverse transcriptase, and in vitro anti-HIV activity. PMID- 14565325 TI - Synthesis and antimalarial activity of novel N6-substituted adenosine derivatives. PMID- 14565326 TI - New lipophilic derivatives of AZT and d4T 5'-phosphonates. AB - 5'-Aminocarbonylphosphonyl and aminocarbonylmethylphosphonyl diesters of AZT and d4T were synthesized as potential anti-HIV agents. PMID- 14565327 TI - Urinary modified nucleosides as tumor markers. AB - Extracts of urinary nucleosides have been sequentially purified and examined by mass spectrometric analysis. Seventeen modified nucleosides have been unequivocally identified and a further five provisionally identified. While several nucleosides were found only in a small number of extracts, the occurrence and levels of others were found to correlate with the tumour type and stage. PMID- 14565329 TI - Metabolic and pharmacological characteristics of the bicyclic nucleoside analogues (BCNAs) as highly selective inhibitors of varicella-zoster virus (VZV). PMID- 14565328 TI - In vitro study of resistance-associated genotypic mutations to nucleoside analogs. AB - In spite of a rather long period of investigations, the problem of HIV drug resistance remains unsolved, and more that, at present HIV-1 mutants resistant to all known nucleoside inhibitors being used in clinical therapy against the human immunodeficiency syndrome are discovered. In this study we selected HIV-1 mutants resistant to the nucleoside inhibitors of HIV reverse transcriptase (NRTI): 3' azido-2',3'-dideoxythymidine (AZT), 5'-phosphit 3'-azido-2',3'-dideoxythymidine (ph-AZT), dideoxyinosine (ddI) and didehydrodeoxythymidine (d4T). Selection of resistant mutants was carried out by gradually increasing of drug concentration in the culture medium during propagation of the HIV-1EVK on fresh MT-4 cells. Phenotypic resistance was defined as an increase in ID50 of 160-fold for AZT, 8 for ph-AZT, 10 for ddI, 7 for d4T. In comparison studies it was determined that the viral resistance to these drugs was appeared variously in a similar conditions and duration of selection. The nucleotide sequences of the RT region of the HIV-1 variants were compared with the HIV-1EVK from "0" passage. For some of selected HIV-1 mutants NRTI resistance mutations were detected. Selected AZT resistant variants contained amino acid substitutions in positions D67A and K70R. Our studies was not revealed substitution at position 75 for ph-AZT resistant variants, whereas substitution at position L214F have been observed in both experiments using AZT and ph-AZT. Selected d4T resistant mutants contained amino acid substitutions in positions N54D and P52R. Selected ddI resistant mutants contained only one amino acid substitution in position P143S. Collection of drug resistant mutants should prove to be a convenient tool for rapid investigations a new antiretroviral agents on cross drug-resistance. PMID- 14565330 TI - Characterization of the catalytic subunit of the human herpesvirus 6 (HHV-6) DNA polymerase expressed in an in vitro transcription/translation assay. PMID- 14565331 TI - Monoval-LdC: efficient prodrug of 2'-deoxy-beta-L-cytidine (L-dC), a potent and selective anti-HBV agent. AB - In order to improve the oral bioavailability of LdC, valinyl esters were prepared as prodrugs. We report here the syntheses of the 3'-mono-, 5'-mono, and 3',5'-di O-valinyl esters of LdC. The comparison of their ease of synthesis, their physicochemical properties, as well as their pharmacokinetic parameters in cynomologus monkeys has revealed 3'-mono-O-valinyl derivative as the most promising of the studied prodrugs. This compound is being developed as a new anti HBV agent. PMID- 14565332 TI - Synthesis of 2'-O-substituted ribonucleosides. AB - An efficient synthesis of 2'-O-substituted ribonucleosides, including 2'-O-TBDMS and 2'-O-TOM protected as well as 2'-O-Me and 2'-O-allyl derivatives is presented. Di-t-butylsilylene group was employed for simultaneous protection of 3'- and 5'- hydroxyl functions of nucleoside on the first step. Subsequent silylation or alkylation of free 2'-OH followed by introduction of suitable protection on the base moiety and removal of cyclic silyl protection gave target compounds in a high yield. PMID- 14565333 TI - Novel base-labile protecting groups for 5'-hydroxy function in solid-phase oligonucleotide synthesis. AB - The 6-(levulinyloxymethyl)-3-methoxy-2-nitrobenzoyl (LMMoNBz) and 2 (levulinyloxymethyl)-5-methoxy-4-nitrobenzoyl (LMMpNBz) groups were developed as novel base-labile protection for the 5'-hydroxy function in solid-phase oligonucleotide synthesis. A comparative study of the LMMoNBz, LMMpNBz and 2 (levulinyloxymethyl)-5-nitrobenzoyl (LMNBz) protecting groups for oligonucleotide synthesis proved strong feasibility for the LMMoNBz group. PMID- 14565334 TI - Progress towards a submonomer synthesis of peptide nucleic acid. AB - We report recent developments in the optimization of a submonomer synthesis of peptide nucleic acid based on the Fukuyama-Mitsunobu reaction. The key steps in the submonomer synthesis are the installation of an appropriately protected 2 aminoethyl group on the alpha-nitrogen of an amino acid and its subsequent acylation with a protected nucleobase derivative. The aggressive alkylation conditions require a scheme of maximal protection for the nucleobases and that is proposed herein for the pyrimidines. PMID- 14565335 TI - Towards a circular bis-peptide nucleic acid. AB - En route to a circular bis-PNA molecule, we have synthesized and characterized the DNA binding of several "clamp"-type bis-PNAs. In order to incorporate charge into a circular PNA, a new linker based on the achiral 2-aminoethylglycine has been used. PMID- 14565336 TI - Nucleobase modified peptide nucleic acid. AB - The Pd0/Cu1 catalyzed cross-coupling of terminal alkynes onto peptide nucleic acid monomers or submonomers bearing iodinated nucleobases has been utilized as a route to base-modified oligomers. Both 5-iodouracil and 5-iodocytosine derivatives undergo the cross-coupling to give the expected products in moderate to good yields. However, depending on the particular substrates and reaction conditions, the cross-coupling may be followed by a ring closing reaction to give the fluorescent furano- and pyrrolo-fused uracil and cytosine derivatives, respectively. PMID- 14565337 TI - A new class of Spiegelmers containing 2'-fluoro-nucleotides. AB - Synthesis of 2'-fluoro-nucleosides from L-arabinose in order to perform the synthesis of 2'-fluoro-Spiegelmers binding to a neuropeptide. PMID- 14565338 TI - Synthesis of an asymmetrically substituted AZA crown ether as metal and amino acid binding site in DNA conjugates. AB - Crown ether 4 as a receptor core for protonated primary amines such as amino acids has been synthesized and incorporated into oligodeoxynucleotides as dangling ends. PMID- 14565340 TI - Structural pre-organization of peptide nucleic acids. AB - Introduction of constraint via chemical bridging in the aegPNA leads to the five or six membered cyclic structures that may contribute towards maintaining the balance between rigidity and flexibility of the PNA backbone. The significant promise of our approach to use the naturally occurring trans-4-hydroxy-L-proline to arrive at different chirally pure cyclic PNA analogs and their DNA binding properties will be presented. PMID- 14565339 TI - Nuclease stability of LNA oligonucleotides and LNA-DNA chimeras. AB - The stabilizing properties of LNA and alpha-L-LNA oligonucleotides against endo- and 3'-exonucleases have been evaluated. PMID- 14565341 TI - Oligomerization aiming at phosphonate analogues of oligoadenylates. AB - This work deals with isopolar, phosphonate-based nucleotide analogues containing a bridging P-C bond instead of the ester P-O linkage. Specifically, starting from activated derivatives 1, 2, and 3, a simple process for preparation of mixtures of short oligomers and their analyses were elaborated. PMID- 14565342 TI - Ribo-, xylo-, and arabino-configured adenine-based nucleoside phosphonates: synthesis of monomers for solid-phase oligonucleotide assembly. AB - Adenine-based, regioisomeric nucleoside phosphonates with ribo, xylo and arabino configuration were synthesized in the protected form suitable for the phosphotriester-like, solid-phase synthesis of oligonucleotides. Phosphonate moiety was protected by 4-methoxy-1-oxido-2-picolyl group and the furanose hydroxyl by the dimethoxytrityl group. PMID- 14565344 TI - A-hydroxyphosphonate oligonucleotides: a promising DNA type? AB - The synthesis of monomers (S)-1, (R)-1 and 2 derived from (5'S)-, (5'R)-2' deoxythymidine-5'-C-phosphonic acids and 2',5'-dideoxythymidine-5'-C-phosphonic acids was elaborated. The protection of the 5'-hydroxyl by the methoxycarbonyl group was a key step of the synthesis. Prepared monomers were used for the solid phase assembly of several types oligothymidylate 15-mers (S)-3, (S)-4, (S)-5, (R) 4 and (R)-5 containing the chiral 3'-O-P-CH(OH)-5'' internucleotide linkage. Their hybridization properties with dA15 and rA15 were studied as well as their resistance against nuclease cleavage. PMID- 14565343 TI - Hybridization properties of 4'-branched oligonucleotides. PMID- 14565345 TI - Isosteric phosphonate pyrrolidine-based dinucleoside monophosphate analogues. AB - A novel alpha- and beta-configured pyrrolidine nucleoside phosphonates in adenine series were synthesized from trans-4-hydroxy-L-proline as starting material. d(ApA) analogues were also prepared and studied with respect to their hybridization properties with polyU. PMID- 14565346 TI - Oligonucleotides containing an acridine group covalently bonded to the nucleotide flanking the 3'-3' phosphodiester junction for alternate strand triple helix formation. AB - Oligonucleotides with a 3'-3' inversion of polarity and containing an acridine group attached to nucleotide base flanking the 3'-3' phosphodiester bon have been synthesized, characterized and used as third strand in alternate triple helix formation. CD melting studies and molecular mechanics calculations have been carried out to investigate these triplex structures. PMID- 14565347 TI - New synthesis of 5-carboxy-2'-deoxyuridine and its incorporation into synthetic oligonucleotides. AB - 5-Carboxy-2'-deoxyuridine is a methyl oxidation product of thymidine. It can be formed by the menadione-mediated photosensitization of thymidine in aerated aqueous solution. Here in we present a new four-step synthesis of the 5-carboxy 2'-deoxyuridine phosphoramidite building block based on the alkaline hydrolysis of 5-trifluoromethyl-2'-deoxyuridine. The phosphoramidite derivative has been incorporated at defined sites into oligonucleotides using the solid phase synthesis approach. PMID- 14565348 TI - Synthesis and hybridization properties of oligonucleotide analogues containing ornithine backbone modified with nucleoalanines. PMID- 14565349 TI - Synthesis of thymine-modified oligonucleotides. AB - Oligonucleotide-resins containing N-nitrothymidine residues yield N3-thymine modified oligonucleotides by reaction with a variety of amines followed by the standard ammonia treatment. PMID- 14565350 TI - 4-Guanidino-2-pyrimidinone nucleobases: synthesis and hybridization properties. AB - N-Alkylated 4-guanidino-2-pyrimidinone-containing nucleosides, in which the guanidine group mimics the double hydrogen bond donor pattern of protonated cytosine, were introduced in polypyrimidine sequences to explore their triple helix forming capabilites. UV and CD melting experiments showed that strands containing these base analogues did not form triplex complexes. PMID- 14565351 TI - Synthesis and DNA binding properties of DNA-PNA chimeras. AB - A systematic study to evaluate the ability of 5'-DNA-3'-p-(N)-PNA-(C) chimeras to form triple helix structures has been undertaken. Preliminary results carried out on a 16-mer chimera with three PNA monomers at the 3'-end showed the formation of a stable DNA-PNA/DNA/DNA triplex, having similar conformational behaviour to a canonical DNA/DNA/DNA triplex. PMID- 14565352 TI - Deoxyribonolactone lesion in DNA: synthesis of fluorinated analogues. AB - Mono- and difluorinated derivatives of 2-deoxyribonolactone were synthesized using diastereoselective Reformatski reaction as a key step. PMID- 14565353 TI - Triplex formation involving 2'-O,4'-C-methylene bridged nucleic acid (2',4'-BNA) with 2-pyridone base analogue: efficient and selective recognition of C:G interruption. AB - For the effective recognition of C:G interruption in homopurine-homopyrimidine duplex DNA, we examined triplex-forming ability and sequence-selectivity of a triplex-forming oligonucleotide (TFO) involving of 2'-O,4'-C-methylene bridged nucleic acid with 2-pyridone base analogue. We found that the modified TFO formed stable triplex with high binding affinity and sequence-selectivity. PMID- 14565354 TI - Pyrrolidine PNA-DNA chimeric oligonucleotides with extended backbone. AB - Cis-D-2-hydroxy-4-thymin-1-yl-pyrrolidine propionic acid unit is used to make PNA DNA dimer block that is incorporated in DNA sequences at selected positions. Since the amide linkage is shorter than phosphodiester linkage, insertion of an extra atom in the backbone with amide linkage seems to be better accommodated for internucleotide distance-complementarity. PMID- 14565355 TI - Piperidinyl peptide nucleic acids: synthesis and DNA-complementation studies. AB - Synthesis of a new six membered PNA analogue by introducing a methylene bridge between beta carbon atom of ethylene diamine and beta' carbon atom of linker to nucleobase. PMID- 14565356 TI - Synthesis and conformational properties of O-beta-D-ribofuranosyl-(1''-2') guanosine and (adenosine)-5''-phosphate. AB - The efficient synthesis of Grp and Arp, minor tRNA components, has been developed. PMID- 14565357 TI - Chemical incorporation of 1-methyladenosine, minor tRNA component, into oligonucleotides. AB - The synthesis of suitably protected 1-methyladenosine derivatives has been developed and its successful chemical incorporation into oligonucleotides was achieved. PMID- 14565358 TI - Oligonucleotides containing disaccharide nucleosides: synthesis, physicochemical, and substrate properties. AB - The efficient synthesis of oligonucleotides containing 2'-O-beta-D-ribofuranosyl (and beta-D-ribopyranosyl)nucleosides, 2'-O-alpha-D-arabinofuranosyl (and alpha-L arabinofuranosyl)nucleosides. 2'-O-beta-D-erythrofuranosylnucleosides, and 2'-O (5'-amino-5-deoxy-beta-D-ribofuranosyl)nucleosides have been developed. PMID- 14565359 TI - Synthesis of oligonucleotide building blocks of 2'-deoxyguanosine bearing a C8 arylamine modification. AB - C8-Arylamine-dG adducts were synthesized by palladium-catalyzed cross-coupling reactions. The corresponding 5'-O-DMTr-3'-O-phosphoramidite-C8-arylamine-dG adducts were synthesized as potential building blocks for the automated synthesis of site-specifically modified oligonucleotides. PMID- 14565360 TI - Alpha-hydroxybenzylphosphonate modified oligonucleotides: synthesis, properties, and a novel route via monomer building blocks. AB - In general, alpha-hydroxybenzylphosphonate modified 2'-deoxyadenosine-thymidine dimer building blocks 1, 2 are utilized for the incorporation into alpha hydroxybenzylphosphonate pro-oligonucleotides. For a universal application of our pro-oligonucleotide concept on biologically relevant oligonucleotides a route for the synthesis of modified monomer building blocks 3 was developed and is presented herein. PMID- 14565361 TI - Determination of the absolute P-configuration of a phthalidyl-phosphonate thymidine-thymidine dimer. AB - Phthalidyl modified oligonucleotide thymidine-thymidine dimer building blocks were synthesized via the H-phosphonate-method. The compounds which are diastereomeric at the phosphorus atom were separated by chromatography and the absolute configuration at the phosphorus atoms was determined using ROE experiments using the corresponding methyl-phosphonates. PMID- 14565362 TI - Improved synthesis of 2'-amino-LNA. AB - 2'-Amino-LNA phosphoramidite (10) was synthesised by means of a new strategy, which is convergent with the synthesis of 2'-oxy-LNA up until a late stage intermediate (1). PMID- 14565363 TI - Synthesis of an oligonucleotide analogue of ethenoadenosine. AB - A phosphoramidite building block derived from 11-carboxy-1,N6-ethenoadenosine has been prepared to be used in a solid supported oligonucleotide synthesis. PMID- 14565364 TI - Conformationally restricted dinucleotides: tandem ring-closing metathesis and hydrogenation approach. AB - Cyclic dinucleotides with saturated connections between a nucleobase and the phosphate are synthesised using a tandem ring-closing metathesis (RCM) and hydrogenation protocol and found to be significantly stabilised towards ammonia. PMID- 14565365 TI - Alpha-LNA (alpha-D-configured locked nucleic acid). AB - Two pyrimidine alpha-LNA nucleoside monomers have been synthesised and incorporated into alpha-configured oligonucleotides. A fully modified mixed alpha LNA sequence displays unprecedented parallel stranded hybridisation with complementary RNA and a remarkable selectivity for RNA over DNA. Modelling shows alpha-LNA:RNA to form an extended duplex with a very broad major groove. PMID- 14565366 TI - xylo-configured oligonucleotides (XNA, xylo nucleic acids): synthesis and hybridization studies. AB - We report synthesis and high-affinity hybridization of fully modified home thymine 2'-deoxy and 2'-deoxy-2'-fluoro xylo nucleic acids. PMID- 14565367 TI - RNase H activation by stereoregular boranophosphate oligonucleotide. AB - A stereoregular all-(Sp)-boranophosphate oligodeoxyribonucleotide (BH3(-)-ODN) 15 mer was synthesized using an enzymatic approach. The BH3(-)-ODN formed a hybrid with the complementary RNA 15-mer and induced RNase H hydrolysis of the RNA strand at ODN concentrations as low as 10 nM at 37 degrees C, but with a lower efficiency than that of its natural phosphodiester analogue. PMID- 14565368 TI - Synthesis of a base-protected alpha-L-LNA guanine nucleoside. AB - Synthesis of (1R,3R,4S,7R)-7-hydroxy-1-hydroxymethyl-3-(2-N-isobutyroylguanin-9 yl)-2,5-dioxabicyclo[2.2.1]heptane (12), a protected alpha-L-LNA guanine nucleoside, has been accomplished using a convergent synthetic strategy starting from 1,2-di-O-acetylfuranose 4. PMID- 14565369 TI - Synthesis and hybridization properties of oligodeoxynucleotides containing 3' deoxy-3'-C-methyleneuridine. AB - 3'-Deoxy-3'-C-methyleneuridine nucleoside 1 has been incorporated into oligodeoxynucleotides. Relative to the unmodified references, oligomers containing nucleoside 1 displayed reduced binding affinities towards complementary DNA and RNA with a tendency towards RNA-selective hybridization. PMID- 14565370 TI - Synthesis of a base-protected xylo-LNA adenine nucleoside. AB - Synthesis of (1S,3R,4R,7R)-7-hydroxy-1-hydroxymethyl-3-(6-N-benzoyl-adenin-9-yl) 2,5-dioxabicyclo[2.2.1]heptane (2), a base-protected xylo-LNA adenine nucleoside, has been accomplished using a convergent synthetic strategy starting from 1,2-di O-acetylfuranose 3. PMID- 14565371 TI - Synthesis of modified RNA-oligonucleotides for structural investigations. AB - RNA exhibits a higher structural diversity than DNA and is an important molecule in biology of life. It shows a number of secondary structures such as duplexes, hairpin loops, bulges, internal loops etc. However, in natural RNA, bases are limited to the four predominant structures U, C, A, and G and so the number of compounds that can be used for investigation of parameters of base stacking, base pairing and hydrogen bond, is limited. We synthesized different fluoromodifications of RNA building blocks: 1'-deoxy-1'-(2,4,6-trifluorophenyl) beta-D-ribofuranose (F), 1'-deoxy-1'-(2,4,5-trifluorophenyl)-beta-D-ribofuranose (M) and 1'-deoxy-1'-(5-trifluoromethyl-1H-benzimidazol-1-yl)-beta-D-ribofuranose (D). Those amidites were incorporated and tested in a defined A, U-rich RNA sequence (12-mer, 5'-CUU UUC XUU CUU-3' paired with 3'-GAA AAG YAA GAA-5') (Schweitzer, B.A.; Kool, E.T. Aromatic nonpolar nucleosides as hydrophobic isosters of pyrimidine and purine nucleosides. J. Org. Chem. 1994, 59, 7238 pp.). Only one position was modified, marked as X and Y respectively. UV melting profiles of those oligonucleotides were measured. PMID- 14565372 TI - Synthesis of dinucleoside boranophosphates by a boranophosphotriester approach. AB - Dinucleoside boranophosphates including four kinds of nucleobases were synthesized by a boranophosphotriester method in good yields. In the present boranophosphotriester method, side-reactions at the nucleobases, which caused by a borane reagent, were completely avoided. PMID- 14565373 TI - Synthesis and binding properties of a homopyrimidine 2',5'-linked xylose nucleic acid (2',5'-XNA). AB - The synthesis and hybridization properties of pyrimidine 2',5'-RNA and 2',5' Xylose Nucleic Acid (2',5'-XNA) are described. PMID- 14565374 TI - Conjugates of oligo(2'-O-methylribonucleotides) with minor groove binders as new sequence-specific agents recognizing both grooves of double-stranded DNA. AB - Design, synthesis and physico-chemical studies of new pyrimidine oligo(2'-O methylribonucleotide) conjugates with one or two oligo(pyrrolecarboxamide) minor groove binders (MGB) are described. PMID- 14565375 TI - Towards a DNA-like duplex without hydrogen bonds. AB - The inverse quadrupolar moments of the phenyl and pentafluorophenyl residues in the base pair P-F5 promotes strong intramolecular stacking interactions in DNA duplexes. The more natural base pairs are replaced by this novel pair the higher the thermodynamic stability of the resulting duplex if they are arranged in an alternating fashion. PMID- 14565377 TI - Fluorinated peptide nucleic acid. AB - The fluorinated olefinic peptide nucleic acid analogue (F-OPA) monomer containing the base thymine was synthesised in 13 steps. PNAs containing this unit were prepared and their pairing properties assessed by means of UV-melting experiments. PMID- 14565376 TI - Pyrrolidino-DNA. AB - We synthesized pyrrolidino-C-nucleosides, incorporated them into oligodeoxynucleotides and investigated their pairing properties. The thermal duplex and triplex stabilities were measured. While triplex formation is destabilized in the case of pyrrolidino-pseudo-U and -T, pyrrolidino-pseudo-iso-C leads to an increase of the Tm value for third strand dissociation. Duplexes are destabilized with all pyrrolidino-C-nucleosides. PMID- 14565378 TI - Transition metal ligands as novel DNA-base substitutes. AB - The base modified nucleoside dBP, carrying a non-hydrogen-bonding non-shape complementary base was incorporated into oligonucleotides (Brotschi, C.; Haberli, A.; Leumann C.J. Angew. Chem. Int. Ed. 2001, 40, 3012-3014). This base was designed to coordinate transition metal ions into well defined positions within a DNA double helix. Melting experiments revealed that the stability of a dBP:dBP base couple in a DNA duplex is similar to a dG:dC base pair even in the absence of transition metal ions. In the presence of transition metal ions, melting experiments revealed a decrease in duplex stability which is on a similar order for all metal ions (Mn2+, Cu2+, Zn2+, Ni2+) tested. PMID- 14565379 TI - New nucleoside analogues for the recognition of pyrimidine-purine inversion sites. AB - A new nucleoside designed to enhance triplex stability has been synthesised in 15 steps starting from sugar 2. This pathway contains the sugar derivative 9 which is a useful intermediate for the introduction of other natural and unnatural bases into the 2'-aminoethoxy nucleoside containing scaffold. PMID- 14565380 TI - Synthetic studies towards a novel, chemical stable, abasic site analogue of DNA. AB - We synthesized the phosphinate 7 via photoaddition of methanol to the alpha, beta unsaturated deoxyribono lactone as the key step, followed by an Arbusov reaction for the introduction of phosphorous. Precursor 7 serves for the synthesis and incorporation into DNA of a novel chemically stable abasic site analogue that might act as an inhibitor for DNA glycosylases. PMID- 14565381 TI - Synthesis of a cyclopentane amide DNA analogue and its base pairing properties. AB - cpa-DNA monomers containing the bases adenine and thymine have been synthesized starting from the known compound 1 in 12 steps. Partially and fully modified cpa thymidine and cpa-adenosine containing oligodeoxynucleotides were synthesized by standard oligonucleotide chemistry. Fully modified homo-cpa-A sequences lead to duplex destabilization by -1.4 degrees C/mod. relative to DNA. As its congener bca-DNA, cpa-DNA prefers left-handed duplex formation where possible. PMID- 14565382 TI - Olefinic peptide nucleic acid (OPA). AB - The olefinic peptide nucleic acid analogues (OPA) monomers containing the bases thymine and adenine were synthesised in 11 steps. Fully modified oligomers containing these units were prepared and their pairing properties assessed by means of UV-melting experiments. PMID- 14565383 TI - Synthesis, biophysical, and biochemical properties of PNA-DNA chimeras. AB - Three PNA-DNA chimeric dimer synthons (tT, upT and uhT, see Sch. 1) have been synthesized in solution and used to make T20-analogue chimeras applying standard solid-phase DNA synthesis protocol. Duplex forming ability of chimeras with dA20 and their hydrolyses by 3'- and 5'-exonucleases (snake venom and bovine spleen phosphodiesterase, respectively) have been investigated. PMID- 14565384 TI - Synthesis of modified DNAs bearing two different fluorescent probes and its application to fluorescence resonance energy transfer. PMID- 14565385 TI - Synthesis and properties of oligonucleotides involving a perylene unit linked to a 2'-deoxyribose residue. AB - We report here the synthesis and binding properties of oligonucleotides involving a perylene unit linked to the anomeric position of a 2'-deoxyribose residue. Both anomers were separated and incorporated separately at either the 5'-end or the internal position of a pyrimidine sequence. In any case the presence of the perylene unit stabilizes the complexes formed with either the single or the double-stranded target. PMID- 14565386 TI - Methylated hexitol nucleic acids, towards congeners with improved antisense potential. AB - In an effort to further improve the hybridisation potential of anhydro-hexitol nucleoside analogues, the 1'-methoxyl and 3'-methoxyl substituents were introduced and evaluated for their antisense potential. In view of the selectivity of pairing with RNA, especially the introduction of a 3'-O-alkyl moiety deserves further study. PMID- 14565387 TI - Oligonucleotides incorporating 7-(aminoalkyn-1-yl)-7-deaza-2'-deoxyguanosines: duplex stability and phosphodiester hydrolysis by exonucleases. AB - Self-complementary [[5'-d(G-C)4]2] and non-selfcomplementary oligonucleotides [5' d(TAG GTC AAT ACT) x 3'-d(ATC CAG TTA TGA)] containing 7-(omega-aminoalkyn-1-yl) 7-deaza-2'-deoxyguanosines (1a-c) (1) and 7-deaza-2'-deoxyguanosine instead of dG were studied regarding their thermal stability as well as their phosphodiester hydrolysis by either 3' --> 5'- or 5' --> 3'-phosphodiesterase studied by MALDI TOF MS. PMID- 14565388 TI - Oligonucleotides incorporating N7-(2'-deoxy-beta-D-erythro pentofuranosyl)isoguanine. AB - The H-phosphonate and the phosphoramidite of N7-2'-deoxyisoguanosine (2) were prepared and incorporated into oligonucleotide duplexes. Their base pairing properties were investigated and compared with those of the parent purine nucleosides. PMID- 14565389 TI - Synthesis and properties of halogenated 7-deaza-2'-deoxyxanthosine and protected derivatives for oligonucleotide synthesis. AB - The 7-bromo- (4a) and 7-iodo- (4b) derivatives of 7-deaza-2'-deoxyxanthosine (5) are prepared. Furthermore, the building blocks 6-8 of 7-deaza-2'-deoxyxanthosine (5) are synthesized and tested for their usage in oligonucleotide synthesis. PMID- 14565390 TI - Synthesis of oligonucleotide prodrugs bearing N-acetyl nucleobases. AB - N-Acetyl oligonucleotides and their prodrugs were synthesized on photolabile solid support. Tm studies showed a decrease of hydridization for N-acetyl A and G and an increase for N-acetyl C. In cells extract, acetyl groups were hydrolysed. PMID- 14565391 TI - Preparation of 6-thioguanosine phosphoramidite for oligoribonucleotide synthesis. AB - 6-Thioguanosine phosphoramidite was prepared, using 2,4-dinitrophenyl as protection group for thio-function, and its stability towards conditions of RNA synthesis was investigated. The results show that the monomer was stable under the conditions of RNA synthesis and suitable for incorporation of thioguanine into oligoribonucleotides. PMID- 14565392 TI - Synthesis and physicochemical properties of 2'-deoxy-2',2''-difluoro-beta-D ribofuranosyl and 2'-deoxy-2',2''-difluoro-alpha-D-ribofuranosyl oligonucleotides. AB - We present procedures for nucleoside and oligonucleotide synthesis, binding affinity (Tm) and structural analysis (CD spectra) of 2'-deoxy-2',2''-difluoro alpha-D-ribofuranosyl and 2'-deoxy-2',2''-difluoro-beta-D-ribofuranosyl oligothymidylates. Possible reasons for the thermal instability of duplexes formed between these compounds and RNA or DNA targets are discussed. PMID- 14565393 TI - Synthesis of oligonucleotide-peptide PEG-conjugated: the EGG (oligonucleotide) chicken (peptide) dilemma? AB - The synthesis of a peptide-PEG-oligonucleotide chimera is compared when starting from the peptide or from the oligonucleotide sequence. PMID- 14565394 TI - The synthesis of 2'-O-methyl G-clamp containing oligonucleotides and their inhibition of the HIV-1 Tat-TAR interaction. AB - We have synthesised a 2'-O-methyl riboside phosphoramidite derivative of the cytosine analogue 9-(2-aminoethoxy)-phenoxazine ('G-clamp') and successfully incorporated it into a series of small steric blocking 2'-O-methyl oligonucleotides targeting the stem-loop region of HIV-1 TAR RNA. The 'G-clamp' containing oligonucleotides show significant increases in binding to a model TAR RNA system when the 'G-clamp' is positioned opposite the loop region. The oligonucleotides also display dose-dependent inhibition of Tat-dependent transcription of an HIV DNA template in HeLa cell nuclear cell extract. PMID- 14565395 TI - Imidazolethyl-phosphoramidate alpha-oligonucleotides. AB - Alpha-ODNs conjugated to imidazole groups via phosphoramidate internucleosidic linkages were synthesized. The presence of the imidazolethyl-phosphoramidate linkage improved the affinity of alpha-ODNs for their nucleic acid targets. PMID- 14565396 TI - Stabilization of DNA double and triple helices by conjugation of minor groove binders to oligonucleotides. AB - New conjugates containing two parallel or antiparallel carboxamide minor groove binders (MGB) attached to the same terminal phosphate of one oligonucleotide strand were synthesized. The conjugates interact with their target DNA stronger than the individual components. Effect of conjugated MGB on DNA duplex and triplex stability and their sequence specificity was demonstrated on the short oligonucleotide duplexes and on the triplex formed by model 16-mer oligonucleotide with HIV polypurine tract. PMID- 14565397 TI - New standards in LNA synthesis. PMID- 14565398 TI - Synthesis and studies of modified oligonucleotides-directed triple helix formation at the purine-pyrimidine interrupted site. AB - Triple helix formation is still restricted to oligopurine-oligopyrimidine double stranded DNA target. Herein we focus on our progress achieved in nucleobase and oligonucleotide modifications area to address the chemical challenge to circumvent the recognition of a purine-pyrimidine base pair interruption in an oligopyrimidine-oligopurine DNA sequence. PMID- 14565399 TI - Synthesis and triplex binding properties of oligonucleotides containing a novel nucleobase. AB - The thiazolo-indole compound 1 bearing the complementary donor-acceptor-donor sites (dad) was designed for specific recognition of an AT inverted base pair in pyrimidine triple helix motif. It was successfully incorporated into 14-mer oligonucleotide using a serinol unit as sugar derivative. The triple helix hybridization studies were examined by means of thermal denaturation experiments with a 26-mer DNA duplex containing the AT inverted base pair. PMID- 14565400 TI - Synthesis of trans-L/D-2-(tert-butoxycarbonyl-aminomethyl)-4-(thymin-1-yl) pyrrolidin-1-yl acetic acid. AB - To delineate the binding preferences of stereochemically divergent pyrrolidine PNAs, synthesis of all four diastreomeric monomers of I and the systematic complexation studies of the resultant PNAs with complementary DNA/RNA is essential. We herein report the synthesis of trans-L/D-2-(tert-butoxycarbonyl aminomethyl)-4-(thymin-1-yl) pyrrolidin-1-yl acetic acids I, their incorporation in PNA oligomers and DNA binding studies will be presented. PMID- 14565401 TI - Stabilization of RNA bulges by oligonucleotides containing 2'-naphthylmethyl-2' deoxytubercidine. AB - A novel nucleoside analogue, 2'-naphthylmethyl-2'-deoxytubercidine, is synthesized and incorporated in oligonucleotides that stabilize bulges in partially complementary RNA. PMID- 14565402 TI - Modified nucleoside triphosphates: synthesis and applications for RNA in vitro selection. PMID- 14565403 TI - A novel and convenient method for the synthesis of free 5'-thiol modified oligonucleotides. AB - The synthesis of free 5'-thiol-modified oligonucleotides using a 4,4',4'' trimethoxytrityl (TMTr)-protected linker and standard Poly-Pak purification has been described. PMID- 14565405 TI - Synthesis of 1,4-anhydro-2-deoxy-D-ribitol derivatives from thymidine. AB - 1,2-Dideoxyribose 5-O-succinate, a component of solid support employed in the synthesis of ribozymes, was synthesized from thymidine. The key step was elimination of nucleobase from 2 to afford glycal 3. A number of catalysts for this reaction were tested, resulting in improved and scaleable synthesis. Hydrogenation of the resulting glycal afforded 1,2-dideoxyribose derivative 4 in a high yield. PMID- 14565404 TI - Synthesis and MS analysis of thiazolium and pyridinium derivatives of peptide nucleic acids (PNAs) and peptides. AB - Sensitivity of ESI-MS analysis of crude PNAs is enhanced using their pyridinium or thiazolium derivatives. Identification of the molecular ion of the product is easier when the label contains bromine, based on the isotope distribution. Study of side reactions, occurred upon the synthesis and/or cleavage, is simple with labelling. Sequencing of non-polar peptides is clear as only a(n) type ions can be observed during their MS/MS analysis. PMID- 14565406 TI - Synthesis and analysis of nucleosides bearing pyrrolepolyamide binding to DNA. AB - An efficient synthesis of adenosine bearing pyrrolepolyamide 1 was achieved by coupling of 3 with 2. The CD spectra obtained at several [ligand ]/[duplex] ratios allowed verification of the formation complex of the DNA duplex [d(CGCAAATTGGC)/d(GCCAATTTGCG)] with 1. PMID- 14565407 TI - Synthesis of 2'-spiro ribo- and arabinonucleosides. AB - Four conformationally restricted bicyclic 2'-spiro nucleosides were synthesised and incorporated into oligonucleotides. These spiro nucleotides induced decreased duplex thermostabilities. PMID- 14565408 TI - Aryl-beta-C-LNA monomers as universal hybridization probes. AB - High-affinity universal hybridization is demonstrated for oligonucleotides containing the pyrenyl-LNA monomer 6b, 2'-O-Me-RNA monomers and LNA monomers. PMID- 14565409 TI - Commercial-scale synthesis of protected 2'-deoxycytidine and cytidine nucleosides. AB - Transformation of 2'-deoxyuridine and uridine analogs to protected 2' deoxycytidine and cytidine analogs has been investigated by two different methods. First, traditional triazolation protocol and second p-nitrophenoxylation method. Our studies conclude that the triazolation method is better and suitable for commercial scale-up. PMID- 14565410 TI - Process development for the synthesis of 5'-O-(4,4'-dimethoxytrityl)-N2 isobutyryl-2'-O-(2-methoxyethyl)-guanosine--a potential precursor for the second generation antisense oligonucleotides: an improved process for the preparation of 2'-O-alkyl-2,6-diaminopurine riboside. AB - An efficient four step process for the preparation of 5'-O-(4,4'-dimethoxytrityl) N2-isobutyryl-2'-O-(2-methoxyethyl)-guanosine 1 was developed. Direct 2'-O alkylation of 2,6-diaminopurine riboside 2 was accomplished via inexpensive and commercially available reagents such as KOH, DMSO and alkyl halides at room temperature in 4-6 hrs. Pure 2'-O-(2-methoxyethyl)-DAPR 3 was isolated by crystallization from methanol. Enzymatic deamination of 3 followed by selective N2-isobutyrylation and 5'-O-dimethoxytritylation furnished desired 1 in high yield and purity. Fully optimized four step synthetic process has been scaled up to the pilot plant level. PMID- 14565411 TI - A novel PNA-monomer for recognition of thymine in triple-helix structures. AB - To expand the triplex recognition repertoire of Nucleic Acids, novel nucleobases that recognize thymine in a T-A base pair are still required. A novel conformationally constrained PNA-monomer (II) capable of binding T in a triplex motif was designed and synthesized in 7 steps starting from commercially available dimethyl 2-oxoglutarate. PMID- 14565412 TI - Synthesis and properties of oligonucleotide chimeras containing 5'-amino-2'-deoxy 2'-fluoroarabinonucleosides. AB - Oligonucleotide analogues comprised of 2'-deoxy-2'-fluoro-beta-D-arabinose units joined via P3'-N5' phosphoramidate linkages (2'F-ANA(5'N)) were prepared for the first time. Among the compounds prepared were a series of 2'OMe-RNA-[GAP]-2'OMe RNA 'chimeras', whereby the "GAP" consisted of DNA, DNA(5'N), 2'F-ANA or 2'F ANA(5'N) segments. The chimeras with the 2'F-ANA and DNA gaps exhibited the highest affinity towards a complementary RNA target, followed by the 5'-amino derivatives, i.e., 2'F-ANA > DNA > 2'F-ANA(5'N) > DNA(5'N). Importantly, hybrids between these chimeras and target RNA were all substrates of both human RNase HII and E. coli RNase HI. In terms of efficiency of the chimera in recruiting the bacterial enzyme, the following order was observed: gap DNA > 2'F-ANA > 2'F ANA(5'N) > DNA(5'N). The corresponding relative rates observed with the human enzyme were: gap DNA > 2'F-ANA(5'N) > 2'F-ANA > DNA(5'N). PMID- 14565414 TI - Synthesis of 2',3'-dideoxy-2'-fluoro-3'-thioarabinothymidine and its 3' phosphoramidite derivative. AB - An efficient method for the synthesis of 5'-O-monomethoxytrityl-2',3'-dideoxy-2' fluoro-3'-thioarabinothymidine [(5'MMT)araF-T(3'SH), (5)] and its 3' phosphoramidite derivative (6) suitable for automated incorporation into oligonucleotides, is demonstrated. A key step in the synthesis involves reaction of 5'-O-MMT-2,3'-O-anhydrothymidine (4) (Eleuteri, A.; Reese, C.B.; Song, Q. J. Chem. Soc. Perkin Trans. 1 1996, 2237 pp.) with sodium thioacetate to give (5' MMT)araF-T(3'SAc) (5) (Elzagheid, M.I.; Mattila, K.; Oivanen, M.; Jones, B.C.N.M.; Cosstick, Lonnberg, H. Eur. J. Org. Chem. 2000, 1987-1991). This nucleoside was then converted to its corresponding phosphoramidite derivative, 6, as described previously ((a) Sun, S.; Yoshida, A.; Piccirilli, J.A. RNA, 1997, 3, 1352-1363; (b) Matulic-Adamic, J.; Beigelman, L. Helvetica Chemica Acta 1999, 82, 2141-2150: (c) Fettes, K.J.; O'Neil, I.; Roberts, S.M.; Cosstick, R. Nucleosides, Nucleotides and Nucl. Acids 2001, 20, 1351-1354). PMID- 14565413 TI - A new synthesis of 9-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)guanine (AraF-G). AB - Interesting and very promising antisense properties of 2'-deoxy-2' fluoroarabinonucleic acids ((a) Wilds, C.J.; Damha, M.J. 2'-Deoxy-2' fluoroarabinonucleosides and oligonucleotides (2'F-ANA): synthesis and physicochemical studies. Nucl. Acids Res. 2000, 28, 3625-3635; (b) Viazovkina, E.; Mangos, M.; Elzagheid, M.I.; Damha, M.J. Current Protocols in Nucleic Acid Chemistry 2002, 4.15.1-4.15.21) (2'F-ANA) has encouraged our research group to optimize the synthetic procedures for 2'-deoxy-2'-fluoro-beta-D arabinonucleosides (araF-N). The synthesis of araF-U, araF-T, araF-A and araF-C is straightforward, (Tann, C.H.; Brodfuehrer, P.R.; Brundidge, S.P.; Sapino, C., Jr. Howell H.G. Fluorocarbohydrates in synthesis. An efficient synthesis of 1-(2 deoxy-2-fluoro-beta-D-arabinofuranosyl)-5-iodouracil (beta-FIAU) and 1-(2-deoxy-2 fluoro-beta-D-arabinofuranosyl)thymine (beta-FMAU). J. Org. Chem. 1985, 50, 3644 3647; Howell, H.G.; Brodfuehrer, P.R.; Brundidge, S.P.; Benigni, D.A.; Sapino, C., Jr. Antiviral nucleosides. A stereospecific, total synthesis of 2'-fluoro-2' deoxy-beta-D-arabinofuranosyl nucleosides. J. Org. Chem. 1988, 53, 85-88; Maruyama, T.; Takamatsu, S.; Kozai, S.; Satoh, Y.; Izana, K. Synthesis of 9-(2 deoxy-2-fluoro-beta-D-arabinofuranosyl)adenine bearing a selectively removable protecting group. Chem. Pharm. Bull. 1999, 47, 966-970) however, the synthesis of the guanine analogue is more complicated and affords poor to moderate yields of araF-G (4) ((a) Elzagheid, M.I.; Viazovkina, E.; Masad, M.J. Synthesis of protected 2'-deoxy-2'-fluoro-beta-D-arabinonucleosides. Synthesis of 2' fluoroarabino nucleoside phosphoramidites and their use in the synthesis of 2'F ANA. Current Protocols in Nucleic Acid Chemistry 2002, 1.7.1-1.7.19; (b) Tennila, T.; Azhayeva, E.; Vepsalainen, J.; Laatikainen, R.; Azhayev, A.; Mikhailopulo, I. Oligonucleotides containing 9-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-adenine and -guanine: synthesis, hybridization and antisense properties. Nucleosides, Nucleotides and Nucl. Acids 2000, 19, 1861-1884). Here we describe an efficient synthesis of araF-G (4) that involves coupling of 2-deoxy-2-fluoro-3,5-di-O benzoyl-alpha-D-arabinofuranosyl bromide (1) with 2-chlorohypoxanthine (2) to afford 2-chloro-beta-araF-I (3) in 52% yield. Nucleoside (3) was transformed into araF-G (4) by treatment with methanolic ammonia (150 degrees C, 6 h) in 67% yield. PMID- 14565415 TI - The effect of universal fluorinated nucleobases on the catalytic activity of ribozymes. AB - Four fluoro modified universal nucleobases have been synthesized. The universal nucleobases 1 and 2, containing a 2,4-difluorobenzene as nucleobase and a 4,6 difluorobenzimidazole, respectively, were chemically incorporated into a selected hammerhead ribozyme sequence which has already been retrovirally expressed as an anti-HIV ribozyme to investigate their effect on the catalytic activity of the ribozymes. The substitution of the natural nucleosides with either 1 or 2 results only in a small decrease of the catalytic activity. The Km value for the monosubstituted ribozyme with a 2,4-difluorobenzene is 309 nM(-1), the corresponding kcat is 2.91 x 10(-3) min(-1). A disubstituted hammerhead ribozyme carrying one of each modification has also been synthesized. For a further stabilization of the ribozyme/substrate complex 2'-(beta-aminoethoxy) modified fluorinated nucleosides 15 and 16 have been developed. PMID- 14565416 TI - Alpha-cycloPNA: 1-aminocylopentane-1-carboxylic acid-derived peptide nucleic acid. AB - All four diastereoisomers of 3-thymine-1-((t)butoxycarbonyl)aminocyclopentane-1 carboxylic acid have been synthesised from (S)-dimethyl malate and thymine monomer 12 has been incorporated into an alpha-cycloPNA oligomer. PMID- 14565417 TI - Oxidation of guanine nucleosides to 4-amidinocarbamoyl-5-hydroxyimidazoles by dimethyldioxirane. AB - Final oxidation products generated from guanosine and 2'-deoxyguanosine by reaction with dimethyldioxirane have been identified as 4-amidinocarbamoyl-5 hydroxyimidazoles. PMID- 14565418 TI - DNA conjugates as novel functional oligonucleotides. AB - Oligodeoxynucleotides with RNA cleavage activity 1) were conjugated with amines and peptides by solid phase fragment condensation (SPFC). It was found that 29 mer DNA enzyme conjugated with spermine at its 5'-end showed higher affinity to the target RNA sequence and 40 times higher activity of cleavage than native DNA enzyme. It is also to be noted that conjugate DNA enzymes showed increased resistance against nuclease digestion. PMID- 14565419 TI - Conformationally restricted chiral peptide nucleic acids derived from azetidines. AB - The initial experiments towards the chemical synthesis of conformationally rigid peptide nucleic acid analogues with azetidine moieties have been described. PMID- 14565420 TI - Control of intracellular delivery of oligonucleotides by conjugation with signal peptides. PMID- 14565421 TI - Aryldiazomethane derivatives as reagents for site specific labeling of nucleic acids at phosphate. AB - An efficient and direct labeling method based on direct alkylation of nucleic acids at phosphates by aryldiazomethane derivatives is described. PMID- 14565422 TI - 1,2-Diol and hydrazide phosphoramidites for solid-phase synthesis and chemoselective ligation of 2'-modified oligonucleotides. AB - The preparation of two novel 2'-O-alkyl phosphoramidites bearing 1,2-diol and hydrazide functions for a chemoselective ligation is described. The former amidite was used to obtain 2'-modified oligodeoxyribonucleotides, which can be later oxidised by NaIO4 to generate 2'-aldehyde oligonucleotides. These were successfully conjugated to acceptor molecules. The latter amidite also showed good coupling yields, but the hydrazide function was demonstrated to be labile under basic deprotection conditions. PMID- 14565423 TI - Total stepwise solid-phase peptide-oligonucleotide conjugate synthesis on macroporous polystyrene. AB - An efficient total stepwise solid-phase synthesis of oligonucleotide-peptide conjugates on a macroporous polystyrene is described. Extending our homoserine linker approach, we prepared a range of fluorescein-labelled conjugates containing one of two different peptides together with oligonucleotides containing 2'-deoxynucleoside or 2'-O-methylribonucleoside phosphodiesters, or gapmers containing 2'-deoxyphosphorothioate sequences flanked by 2'-O-methyl wings. PMID- 14565424 TI - Synthesis of 2'-modified oligonucleotides containing aldehyde or ethylenediamine groups. AB - Oligonucleotides carrying 2'-aldehyde groups were synthesized and coupled to peptides containing an N-terminal cysteine, aminooxy or hydrazide group to give peptide-oligonucleotide conjugates in good yield. The synthesis of a novel phosphoramidite reagent for the incorporation of 2'-O-(2,3-diaminopropyl)uridine into oligonucleotides was also described. Resultant 2'-diaminooligonucleotides may be useful intermediates in further peptide conjugation studies. PMID- 14565425 TI - Studies on the stability of dinucleoside H-phosphonates. AB - The stability of two dinucleoside H-phosphonates under various conditions is reported. PMID- 14565427 TI - More efficient photolithographic synthesis of DNA-chips by photosensitization. PMID- 14565426 TI - The cleavage of the triphosphate bridge of a model for the 5'-cap mRNA promoted by dinuclear bicyclic complexes with metal ions. AB - Dinuclear bicyclic complexes, which have two active centers, can significantly promote the hydrolysis of the triphosphate bridge in ApppA, a 5'-cap model compound. PMID- 14565428 TI - Nanoparticles and DNA probes based on DNA-fluorophore conjugates. PMID- 14565429 TI - Novel photocleavable universal support for oligonucleotide synthesis. AB - A novel photocleavable universal support for the automated solid phase synthesis of oligonucleotides is described. The linker between the growing oligonucleotide chain and CPG support contains a nucleophilic amine protected with a photocleavable group. On exposure to UV light, this group is detached and the free amine affords cleavage of the oligonucleotide from the support. The use of long wavelength UV light avoids damage to the DNA. PMID- 14565431 TI - Diastereocontrolled synthesis of phosphorothioate DNA via an oxazaphospholidine approach using a novel class of activators, dialkyl(cyanomethyl)ammonium salts. AB - Dialkyl(cyanomethyl)ammonium salts 1 were synthesized and used as a novel class of activators for the stereospecific condensations of diastereopure nucleoside 3' O-oxazaphospholidines with a nucleoside. This new oxazaphospholidine method could efficiently produce both (Rp)- and (Sp)-dinucleoside phosphorothioates. PMID- 14565430 TI - Stability, vigor, and inherent versatility of novel amino linker and spacer phosphoramidites. AB - Novel amino linker and spacer phosphoramidites were synthesized from methoxyoxalamido (MOX) percursors possessing a secondary hydroxyl, which when phosphitylated endowed stability to the corresponding phosphoramidites. The synthetic strategy is robust, and the chemistry is reactive towards a variety of primary aliphatic diamines and amino alcohols to produce distinctly unique phosphoramidites. The selection of building blocks determines the length and physico-chemical properties of the phosphoramidite tethering arms, and the synthesis can be specifically tailored to suit individual requirement. PMID- 14565432 TI - Understanding high diastereomeric discrimination in formation of oligoribonucleotide phosphorothioate linkages: the first study of pKa-dependent activation in solid-supported coupling of 2'-O-substituted ribonucleoside phosphoramidites. AB - Activation of 2'-O-substituted ribonucleoside phosphoramidites with various activators during solid-supported synthesis of phosphorothioate oligonucleotides was studied. The Rp:Sp diastereomeric composition of resulting phosphorothioate linkage dependent on pKa of activator utilized for coupling. PMID- 14565433 TI - Synthesis of high quality phosphorothioate oligonucleotides as antisense drugs. Use of I-linker in the elimination of 3'-terminal phosphorothioate monoesters. AB - Detritylation of a 5'-O-DMT-2'-deoxyadenosine moiety attached to solid support under acidic condition leads to depurination during oligonucleotide synthesis. Deprotection followed by reversed phase HPLC purification leads to desired oligonucleotide contaminated with significant levels of 3'-terminal phosphorothiaote (3'-TPT) monoester (n-1)-mer. However, it is demonstrated that attachment of dA nucleoside through its exocyclic amino group to solid support leads to substantial reduction of 3'-TPT formation thereby improving the quality of oligonucleotide synthesized. PMID- 14565434 TI - Highly efficient synthesis of peptide- and carbohydrate-oligonucleotide conjugates using chemoselective oxime and thiazolidine formation. PMID- 14565435 TI - Reaction mechanism for the diastereocontrolled synthesis of phosphorothioate DNA by the oxazaphospholidine approach. AB - Reaction mechanisms for the diastereoselective synthesis of deoxyribonucleoside 3'-cyclic phosphoramidites as well as dinucleoside phosphite intermediates by the oxazaphospholidine approach were analyzed by means of ab initio molecular orbital calculations at the HF/6-31G* level. These reactions are essential for the diastereoselective synthesis of phsophorothioate DNA. PMID- 14565436 TI - Convenient solid-phase method of introduction of sulfhydryl groups into oligodeoxyribonucleotides via 2'-O-carbamate linkage. PMID- 14565437 TI - Novel biotin phosphoramidites with super-long tethering arms. AB - A number of novel biotin phosphoramidites, possessing exceptionally long and uncharged tethering arms, were synthesized from methoxyoxalamido (MOX) and succinimido (SUC) precursors. Included among these monomers is a uridine derivative with the biotin moiety attached through the 2'-position. Some of these phosphoramidites were used to make 5'-biotinylated primers, which were applied in direct sequencing of genomic DNA and capture of Sanger fragment pools. PMID- 14565438 TI - Synthesis of novel tyrosinyl FRET cassettes, terminators, and their potential use in DNA sequencing. AB - Fluorescence resonance energy transfer (FRET) dye labeled cassettes and terminators with one or more donor dyes (fluorescein) and acceptor dye (rhodamine dyes) with benzofuran or tyrosine linker moieties were synthesized. These terminators were evaluated for their energy transfer and DNA sequencing potential using thermostable DNA polymerase. PMID- 14565439 TI - Novel synthesis of 2'-O modified oligonucleotides by solid phase fragment condensation. PMID- 14565440 TI - Synthesis of DNA conjugates by solid phase fragment condensation. AB - Development of a novel method for the synthesis of DNA conjugates is described. Oligonucleotides were successfully conjugated with a variety of functional molecules on a solid phase (Solid Phase Fragment Condensation) using an amino, a hydroxyl, a thiol, and a carboxyl group. DNA-peptide conjugate was obtained as a pure from by a single RPHPLC purification approximately in 20% yield. Moreover, it was demonstrated that the present method was effective for the preparation of conjugate molecules, DNA-sugar, DNA-polyamine, DNA-lipid and so on. The study to create new intelligent DNAs by accumulation various biofunctions on the molecule by SPFC is now in progress in our laboratory. PMID- 14565441 TI - Direct regioselective enzymatic acylation of nucleosides: building-blocks for the solution phase synthesis of oligonucleotides. AB - The synthesis of 3'- and 5'-O-levulinyl nucleosidic monomers through enzymatic acylation with acetonoxime levulinate is demonstrated. The acylation process takes place in one-step and use of expensive reagents, such as DMTrCl is avoided. The regioselectivity of the procedure makes it very convenient for acylated monomers required for solution phase synthesis of oligonucleotides. PMID- 14565442 TI - Studies towards synthesis of dinucleoside arylphosphonates with metal complexing properties. AB - An efficient and stereospecific synthesis of dinucleoside 4'-(2,2':6',2'' terpyridyl)phosphonate 2 and 5-(2,2'-bipyridyl)phosphonate 3 via a palladium(0) cross coupling strategy has been developed. PMID- 14565443 TI - 9-Fluorenemethyl H-phosphonoselenoate--a versatile reagent for transferring an H phosphonoselenoate group. AB - This paper expands the available methods for preparation of H-phosphonoselenoate using a new reagent, 9-fluorenemethyl H-phosphonoselenoate. PMID- 14565444 TI - Chemoselectivity in oxidative coupling of bifunctional nucleophiles with dinucleoside H-phosphonate and dinucleoside H-phosphonothioate diesters. AB - Chemoselectivity and stereospecificity of iodine mediated oxidative couplings using separate diastereomers of dinucleoside H-phosphonate and H-phosphonothioate with various N- and O-binucleophiles were investigated. PMID- 14565445 TI - Negatively charged, dye labeled-dideoxynucleotides for "direct-load" DNA sequencing. AB - A four-color set of negatively charged, single dye as well as energy transfer dye labeled-ddNTPs were synthesized and evaluated in combination with a novel polymerase in a "direct-load" DNA sequencing, obviating the laborious and time consuming post-reaction work-up. PMID- 14565446 TI - Design, synthesis, and in vitro evaluation of APIO analogue of neplanocin A. AB - A novel apio analogue of neplanocin A was efficiently synthesized from D-ribose via stereoselective aldol-retroaldol reaction for introducing hydroxymethyl group and RCM reaction for synthesizing carbocycle, and its inhibitory activity against SAH hydrolase was assayed. PMID- 14565447 TI - A new concept for DNA-arrays. AB - We will insert a cleavage site in an oligodeoxynucleotide, which can be used for a selective and quantitative cleavage. For that reason we synthesized the four 5' S-(4,4'-dimethoxytrityl)-mercapto-2'-deoxynucleotide-3'-O-(2 cyanoethoxydiisopropylamino)-phosphites (5a-d). The cleavage of P-S and C-S bonds is described (Mag, M.; Lucking, S.; Engels, J.W. Synthesis and selective cleavage of an oligodeoxy-nucleotide containing a bridged internucleotide 5' phosphorthioate linkage. Nucleic Acids Res. 1991, 19 (7), 1437-1441; Marriott, J.H.; Mottahedeh, M.; Reese, C.B. 9-(4-methoxyphenyl)xanthen-9-thiol: A useful reagent for the preparation of thiols. Tetrahedron Lett. 1990, 31 (51), 7485 7488; Divakar, K.J.; Mottoh, A.; Reese, C.B.; Shanghvi, Y.S. Approaches to the synthesis of 2' thio analogues of pyrimidine ribosides. J. Chem. Sc., Perkin Trans. 1 1990, 969-974). The oligodeoxynucleotides with an achiral bridged 5' phosphorothioate linkage 5'-O-P-S-3' are synthesized by the phosphoramidite procedure. PMID- 14565448 TI - Molecular recognition of quadruplex DNA by quinacridine derivatives. AB - The interaction of monomeric and dimeric quinacridines with quadruplex DNA has been investigated using a variety of biophysical methods. Both series of compounds were shown to exhibit a high affinity for the G4 conformation with two equivalent binding sites. As shown from the SPR and dialysis experiments the macrocyclic dimer appears more selective than its monomeric counterpart. PMID- 14565449 TI - Photoadduct leading to crosslinking in Ru(II)-derivatized oligonucleotides. PMID- 14565450 TI - Properties of conjugate DNA enzymes. PMID- 14565451 TI - Novel method for the covalent immobilization of oligonucleotides via Diels-Alder bioconjugation. AB - The synthesis of cyclohexadiene and maleimide derivatives and their use for the functionalization of oligonucleotides and the coating of glass surfaces is reported. A method for the covalent attachment of diene or maleimide modified oligonucleotides to the coated glass surfaces via aqueous Diels-Alder reactions is presented. PMID- 14565452 TI - Fluorescent 2-pyrimidinone nucleoside in parallel-stranded DNA. AB - Stretches of parallel-stranded (ps) double-helical DNA can arise within antiparallel-stranded (aps) Watson-Crick DNA in looped structures or in the presence of sequence mismatches. Here we studied an effect of a pyrimidinone-G (PG) base pair on the stability and conformation of the ps DNA to explore whether P is useful as a structural probe. PMID- 14565453 TI - Cross-linking of Escherichia coli formamidopyrymidine-DNA glycosylase to DNA duplexes containing photoactivatable phenyl(trifluoromethyl)diazirine groups. AB - New reactive analogs of substrates for DNA repair enzyme E. coli Fpg protein containing the residues of 8-oxoguanine and photoactivatable phenyl(trifluoromethyl)diazirine groups were synthesized. Their substrate properties were investigated. Using photocross-linking technique, we established the presence of contacts of two nucleosides located near the oxoG with amino acids from the Fpg protein. The cross-linking efficiency achieved 10%. PMID- 14565454 TI - Oligoribonucleotides with functionalized nucleobases as new modifiers of biopolymers. AB - Synthesis and evaluation of hybridization and modification abilities of the new types of photoactivatable oligoribonucleotide conjugates are presented. PMID- 14565455 TI - Evaluation of capillary HPLC/mass spectrometry as an alternative analysis method for gel electrophoresis of oligonucleotides. AB - A method has been developed to monitor the enzymatic incorporation of nucleotides in DNA by electrospray HPLC mass spectrometry. The main advantages of mass spectrometry over electrophoresis are the ability to directly characterize the reaction products and the shorter analysis time. PMID- 14565456 TI - Structural studies on DNA triple helix formed by intronic GAA triplet repeat expansion in Friedreich's ataxia. PMID- 14565457 TI - Catalytic DNA and RNA for targeting MDR1 mRNA. AB - Design, synthesis and properties of catalytic NAs for targeting MDR1 mRNA are reported. PMID- 14565458 TI - Ability of adenosine-2'(3')-deoxy-3'(2')-triphosphates and related analogues to replace ATP as phosphate donor for all human and Drosphila melanogaster deoxyribonucleoside kinases. AB - Six non-conventional adenosine-2'- and 3'-triphosphate analogues of ATP were tested as potential phosphate donors for all four human, and D. melanogaster, deoxyribonucleoside kinases. With dCK (only dAdo as acceptor), TK1, TK2 and dNK only 3'-deoxyadenosine-2'-triphosphate was an effective donor (5-60% that for ATP). With dCK (dCyd as acceptor) and dGK (dGuo as acceptor), sharing 45% sequence identity, donor activities ranged from 13 to 119% that for ATP. Products were 5'-phosphates. In some instances, kinetics are dependent on the nature of the acceptor, and donor and acceptors properties are mutually interdependent. Results are highly relevant to studies on the modes of interaction with the enzymes, and to interpretations of reported crystal structures of dCK and dNK with bound ligands. PMID- 14565459 TI - 5'-O-fluorosulfonylbenzoyl esters of purine nucleosides as potential inhibitors of NTPase/helicase and polymerase of Flaviviridae viruses. AB - Synthesis and interactions of guanosine, inosine and ribavirin 5' fluorosulfonylbenzoyl esters with hepatitis C virus (HCV) and Flaviviruses NTPase/helicase and polymerase are described. PMID- 14565460 TI - Intersubunit interactions in human cytidine deaminase. AB - In order to design new efficient cytidine based drugs, an intersubunit interactions study related to the active site has been performed on the wild-type cytidine deaminase (CDA) and on the mutant enzyme F137W/W113F. F137 is the homologous to the Bacillus subtilis CDA F125 involved in the subunit interactions. In presence of the dissociating agent SDS, wild-type human CDA dissociate into enzymatically inactive monomers without intermediate forms via a non-cooperative transition. Extensive dialysis or dilution of the inactivated monomers restores completely the activity. The presence of the strong human CDA competitive inhibitor 5-fluorozebularine disfavour dissociation of the tetramer into subunits in the wild-type CDA but not in mutant enzyme F137W/W113F. PMID- 14565461 TI - Human cytidine deaminase: understanding the catalytic mechanism. AB - In the absence of an experimentally elucidated three-dimensional structure of the human CDA, we built an homology model of this enzyme starting from the crystal structure of its E. coli homologous. Furthermore, we docked in the active site alternatively the substrate, the intermediate or the product. By means of molecular dynamics simulations, we determined the topology of the active site, identifying the amino acids involved in the catalytic mechanism, and outlining the central role played by E67. PMID- 14565462 TI - Nicotinamide adenine dinucleotide a unique compound for theoretical and synthetic model studies: chirality as source for high stereospecificity. AB - A dynamic model is given for the hydride transfer of the redox couple NAD+-NADH with model systems and quantum chemical calculations. PMID- 14565463 TI - The biochemical significance of parallel DNA duplexes. AB - Structural and synthetic model are given for (modified) parallel DNAs with non Watson and Crick duplex formation. PMID- 14565464 TI - Partial molar volumes of mRNA 5' cap analogues. AB - Partial molar volumes in aqueous solution of eleven selected 7-methylguanine cap analogues and their guanine counterparts were determined by means of density measurements. Hydrophobicity of the investigated compounds regarding their structural features was analysed within the framework of the solute-solvent interaction model, based on the relative density of the molecular solvation shell. PMID- 14565465 TI - Thermodynamics of 7-methylguanosine cation stacking with tryptophan upon mRNA 5' cap binding to translation factor eIF4E. AB - All eukaryotic nuclear transcribed mRNAs possess the cap structure, consisting of 7-methylguanosine linked by the 5'-5' triphosphate bridge to the first nucleoside. The goal of the present study is to dissect the enthalpy and entropy changes of association of the mRNA 5' cap with eIF4E into contributions originating from the interaction of 7-methylguanosine with tryptophan. The model results are discussed in the context of the thermodynamic parameters for the association of eIF4E with synthetic cap analogues. PMID- 14565466 TI - Recognition of cyclonucleoside lesions by the Lactococcus lactis FPG protein. AB - Several purine and pyrimidine cyclonucleosides were found to be not recognized by several Escherichia coli and yeast DNA N-glycosylases. Interestingly, a non covalent complex was observed between the Lactoccocus lactis formamidopyrimidine DNA glycosylases (Fpg-Ll) and the cyclonucleosides. This may provide new information on the mechanism involved in the activity of the latter enzyme. PMID- 14565467 TI - Interactions of potent multisubstrate analogue inhibitors with purine nucleoside phosphorylase from calf spleen--kinetic and spectrofluorimetric studies. AB - Dissociation constants and stoichiometry of binding for interaction of trimeric calf spleen purine nucleoside phosphorylase with potent multisubstrate analogue inhibitors were studied by kinetic and spectrofluorimetric methods. PMID- 14565468 TI - Triplex formation involving 2'-O,4'-C-methylene bridged nucleic acid (2',4'-BNA) with 1-isoquinolone base analogue: efficient and selective recognition of C:G interruption. AB - For the effective recognition of C x G interruption in homopurine-homopyrimidine duplex DNA, we examined triplex-forming ability and sequence-selectivity of a triplex-forming oligonucleotide (TFO) involving of 2'-O, 4'-C-methylene bridged nucleic acid with 1-isoquinolone base analogue. We found that the modified TFO formed stable triplex with high binding affinity and sequence-selectivity. PMID- 14565469 TI - Inhibition of vertebrate telomerases by the triphosphate derivatives of some biologically active nucleosides. AB - In order to clarify the effect of the base moiety of nucleotide analogs on telomerase inhibition, triphosphate derivatives of biologically active nucleosides, 3'-azido-3'-deoxythymidine (AZT), 2'-deoxy-2' fluoroarafuranosylthymine (FaraT), acycloguanosine (ACG) and their guanine or thymine counterparts (AZdG, FaraG and ACT, respectively) were investigated. In all of the present cases, guanine derivatives showed more potent inhibition than their thymine counterparts. PMID- 14565470 TI - Genosnip: SNP genotyping by MALDI-TOF MS using photocleavable oligonucleotides. AB - A photocleavable o-nitrobenzyl CE phosphoramidite building-block was synthesised and incorporated within oligonucleotides. After allele-specific primer extension, desalting was performed using genostrep purification plates. Release of the SNP information containing part through photocleavage created shortened molecules that are easily accessible for MALDI-TOF analysis. Additionally, incorporation of mass modified nucleosides enables flexible design of multiplex genotyping. PMID- 14565471 TI - MALDI-TOF mass spectrometry as a powerful tool to study enzymatic processing of DNA lesions inserted into oligonucleotides. AB - MALDI-TOF mass spectrometry measurements, coupled with either exonuclease or DNA N-glycosylases digestions of lesion-containing oligonucleotides, were used to assess biochemical features of several oxidative DNA damage. The latter analytical approach was shown to be an informative and efficient alternative technique to conventional electrophoresis and chromatographic analyses. PMID- 14565472 TI - New reagent for protein-DNA contacts footprinting. AB - We have found, that the reaction of o-bromobenzoic acid with Cu2+ ions can be used as a source of activated oxygen species capable of cleaving DNA. Possibility to apply this reaction for footprinting the nucleosome core in the reconstituted chromatin was demonstrated. PMID- 14565473 TI - Effect of putrescine and PEG on a structural transition of DNA G-quadruplex. PMID- 14565474 TI - Synthesis and energy transfer efficiency of FRET terminators derived from different linkers. AB - A number of different energy transfer dye labeled-cassettes were synthesized using aminoacid based trifunctional linkers and coupled to the propargylamino substituted dideoxynucleoside-5'-triphosphates (ddNTPs). These terminators were evaluated for their energy transfer efficiency and DNA sequencing potential using thermostable DNA polymerase. PMID- 14565476 TI - Suppression of exonucleolytic degradation of double-stranded DNA and inhibition of Exonuclease III by PNA. AB - Degradation of double-stranded DNA by Exonuclease III in the presence of complementary anti-parallel PNA was studied. It was found for the first time that the PNA suppresses the degradation of dsDNA in a sequence-specific manner as well as inhibits the activity of Exonuclease III in a non-specific way. PMID- 14565475 TI - Branchpoint sugar stereochemistry determines the hydrolytic susceptibility of branched RNA fragments by the yeast debranching enzyme (YDBR). AB - A series of branched RNAs (Y-shaped) related to yeast pre-mRNA splicing intermediates were synthesized incorporating both natural (i.e., ribose) and non natural (i.e., arabinose, xylose and acyclic nucleoside) branchpoints in order to examine the effect of sugar conformation and phosphodiester configuration on yDBR hydrolytic efficiency. The results indicate that 2'-phosphodiester scission with yDBR occurs only with a cis-arrangement of phosphate groups at the branchpoint (i.e., ribose) thereby discriminating between all other configurations. PMID- 14565477 TI - Antisense knockdown of PKC-alpha using LNA-oligos. AB - Full-length and 4 nucleotides truncated Locked Nucleic Acid (LNA) modifications of ISIS 3521 were compared for antisense properties in a cellular assay. ISIS 3521 is a 20-mer phosphorothioate designed to hybridise to human protein kinase C alpha (PKC-alpha) mRNA and is currently submitted to clinical trials against cancer. We report that LNA can potentate this antisense oligo and retain the antisense potential with shorter oligos. PMID- 14565478 TI - Covalent coupling of a PIM-1 oncogene targeted PNA with an antennapedia derived peptide. AB - Peptide nucleic acids (PNA) are promising antisense molecule for blocking gene expression in cell culture or in vivo. Nevertheless because they are poor efficient to pass the cellular membrane, it is necessary to use a vectorisation agent to observe an inhibitory effect. We describe the coupling of the rhodamine labeled 17-mer antisense PNA to a fusogenic peptide from antenapedia via S-S linkage, the studies of the penetration of this complex into fibroblast cells and its inhibitory effect on pim1 targeted protononcogene. PMID- 14565479 TI - Inhibition of gene expression by peptide nucleic acids in cultured cells. AB - We have tested in cultured cells the capacity of antisense and antigene PNAs to inhibit, in a sequence specific manner, the expression of oncogenes in leukaemia and pancreatic carcinoma cells. The results observed appeared promising and suggest that PNA may play in the future an important role in targeting disease related genes. PMID- 14565480 TI - 2'-O,4'-C-ethylene-bridged nucleic acid (ENA) for effective antisense formation. AB - ENA antisense oligonucleotides for vascular endothelial growth factor (VEGF) mRNA were synthesized and evaluated in A549 lung cancer cells. It was found that the VEGF ENA-antisense inhibited not only the expression of VEGF, but also the expression of three genes, which were found in Genbank by BLAST and Clustal W search and considered likely to bind to the VEGF ENA-antisense. These results indicate that ENA-antisense oligonucleotides act in a sequence-specific manner, and could be used as effective antisense drugs. PMID- 14565481 TI - Synthesis and antitumor activity of N-sulfonyl derivatives of nucleobases and sulfonamido nucleoside derivatives. AB - The introduction of sulfonamido group on the C-2 position of pyrimidine nucleosides was achieved by ring opening of 2,2'- and 2,3'-anhydronucleosides. N sulfonyl derivatives of nucleobases and sulfonamido derivatives of nucleosides were assayed for in vitro antitumor activity. PMID- 14565482 TI - Novel short oligonucleotide conjugates as inhibitors of human telomerase. AB - A series of oligonucleotide conjugates were designed and synthesized as novel inhibitors of human telomerase. These compounds contain a relatively short (6-7 mer) oligonucleotide domain, with an N3'-->P5' phosphoramidate (np) or thio phosphoramidate (nps) backbone, targeted to the template region of the RNA component of the enzyme and various pendant groups attached to either their 5'- or preferably to the 3'-termini. The most potent compounds in the series inhibited telomerase with low nM IC50 values in biochemical assays whereas the cognate oligonucleotides without the pendant groups were significantly less active having IC50 values 100-1000-fold higher. PMID- 14565483 TI - Specific inhibition of hepatitis C viral gene expression by non-polar (phenylalkyl)phosphonates. AB - Different phenylalkyl backbone modified antisense oligonucleotides complementary to the Hepatitis C virus (HCV) RNA nucleotides 326-342 were synthesized. The lipohilic character of modified oligonucleotides was determined from RP-HPLC retention times. The inhibitory effect of these antisense oligonucleotides on HCV gene expression was analyzed in an in vitro test system. PMID- 14565484 TI - Endcaps for stabilizing short DNA duplexes. AB - The syntheses of endcaps for covalently linking the 3' and 5' hydroxyl groups of blunt end double-stranded DNA are described. Endcap diols were converted into DMTr protected phosphoramidites and incorporated between nucleotides 4 and 5 of a self-complementary octamer. The stabilizing effect of the endcaps on duplex DNA was determined by Tm experiments on the self-complementary octamer. PMID- 14565485 TI - Diastereomeric process control in the synthesis of 2'-O-(2-methoxyethyl) oligoribonucleotide phosphorothioates as antisense drugs. AB - Coupling of 2'-O-methoxyethylsubstituted nucleoside phosphoramidites to 5' hydroxyl group of a nucleoside or nucleotide on solid support is under stereochemical process control and is independent of scale, concentration, synthesizer, ratio of amidite diastereomers, solid support etc. However, activators and phosphate protecting groups do play a role in influencing the ratio of phosphorothioate diesters obtained by sulfurization of phosphite triesters. PMID- 14565486 TI - Universal labeling chemistry for nucleic acid detection on DNA-arrays. AB - We show here a new and efficient aqueous chemistry for labeling of any class of nucleic acids for their detection on DNA chip. The labels contain a diazo function as reactive moiety and biotin as detectable unit. The highly selective reaction of diazo group on the phosphate does not disrupt base pairing recognition and hybridization specificity. PMID- 14565487 TI - Incorporation of (alpha-P-borano)-2',3'-dideoxycytidine 5'-triphosphate into DNA by drug-resistant MMLV reverse transcriptase and Taq DNA polymerase. AB - The Rp-stereoisomer of 5'-(alpha-P-borano)triphosphates of 2'-deoxycytidine (Rp dCTPalphaB) and 2',3'-dideoxycytidine (Rp-ddCTPalphaB) were synthesized. Their steady-state kinetics of incorporation by ddNTP-resistant enzymes, e.g., MMLV reverse transcriptase (RT) and Taq DNA polymerase, were investigated and compared with incorporation of dCTP and ddCTP. The alpha-boranophosphate substitution in ddCTP results in a 28-fold increase in efficiency of incorporation of the Rp ddCTPalphaB isomer by MMLV RT, yet has minimal effect on the efficiency of incorporation by Taq DNA polymerase. PMID- 14565488 TI - Synthesis of 2'-deuterio and 3'-deuterio cytidine 5'-diphosphate. AB - 2'-2H- and 3'-2H-CDP were synthesized from 5'-MMT-3'-O-TBDMS and 2',5'-O-diTBDMS cytidine derivatives, respectively, by oxidation followed by acidic removal of 5' protection, reduction with [NaBD(OAc)3] and finally displacement of a tosyl group by pyrophosphate. PMID- 14565489 TI - Determination of RNA tertiary structure by electron paramagnetic resonance spectroscopy (EPR). PMID- 14565490 TI - Synthesis and recognition by DNA polymerases of a reactive nucleoside for DNA diversification. AB - The synthesis of 1-(2-deoxy-beta-D-erythro-pentofuranosyl)imidazole-4-hydrazide having the features of an ambigous base is reported. The recognition of the analogue by DNA polymerases as an incoming triphosphate as well as a template base was investigated. The mutagenic properties was evaluated by PCR. The potential of this new monomer for DNA diversification is illustrated by the reactivity of the nucleobase towards various aldehydes. PMID- 14565491 TI - Conformationally restricted 2-substituted wyosine derivatives. 1H, 13C, and 15N NMR study. AB - It was found by 1H, 13C and 15N NMR study that substitution of 4,9-dihydro-4,6 dimethyl-9-oxo-3-(2',3',5'-tri-O-acetyl-beta-D-ribofuranosyl) imidazo [1,2 a]purine (wyosine triacetate, 1) at C2 position with electronegative groups CH30 and C6H5CH2O results in a noticeable electron distribution disturbance in the "left-hand" imidazole ring and a significant increase in the North conformer population of the sugar moiety. PMID- 14565492 TI - 31P NMR studies on the putative chemoselective activation of nucleoside-3' thiophosphate-O-esters. AB - P-diastereomerically pure O-esters of N(Bz)-5'-DMT-dA-3'-monothiophosphate, having charged S=P-O(-) moiety, have been synthesized. Chemoselectivity of their activations by formation of different mixed anhydrides, followed by couplings with N(Bz)-3'-levulinyl-dA, were studied by 31P NMR spectroscopy. PMID- 14565493 TI - 1H-NMR study of the quadruplex [d(TGGGT)]4 containing a modified thymine. AB - A NMR structural study of quadruplex [d(TGGGT)]4 containing a modified thymine is reported. The three dimensional structure of the complex is very similar to those of other parallel stranded quadruplexes. The modified thymines (T*) are able, at least in the minimised structures, to form a tetrad containing extra H-bonds through the hydroxyl groups. Nevertheless, in this new tetrad the modified thymines are slightly open towards the solvent respect to the unmodified T tetrad. PMID- 14565494 TI - PNA-DNA chimeras forming quadruplex structures. AB - 1H-NMR, CD, and UV spectroscopy have been used to investigate the structure of PNA/DNA chimeras forming quadruplex structures. In particular, we synthesized 5'TGGG3'-t (1) and 5'TGG3'-gt (2), where lower and upper case letters indicate PNA and DNA residues, respectively. CD spectrum and all NMR data of (1) are typical of quadruplexes involving four parallel strands. UV melting profile of (1) indicates that its thermal stability is quite similar to that observed for the reference structure [d(TGGGT)]4. 1H-NMR spectrum for 5'TGG3'-gt (2) shows that this oligonucleotide is not able to fold into a single, well-defined species. PMID- 14565495 TI - Synthesis of 2-deoxy-beta-D-ribose 1-phosphate, NMR comparison and its enzymatic activity for structural elucidation of synthetic alpha-isomer. AB - 2-Deoxy-beta-D-ribose 1-phosphate (1) was synthesized in a stereoselective manner and isolated with no detectable contamination by its alpha-isomer (4). Explicit configuration of 4 was first determined by NMR comparison with 1 judging from NOE results and their coupling constants. Natural purine nucleoside phosphorylase (PNPase) did not recognize 1 and gave no products such as alpha- or beta deoxynucleosides. PMID- 14565496 TI - Structure of 2',5'-linked tetraribonucleotide loops: a novel RNA motif. AB - We report on the three dimensional structure of an RNA hairpin containing a 2',5' linked tetraribonucleotide loop, namely, 5'-rGGAC(UUCG)GUCC-3' (where UUCG = U(2'p5')U(2'p5')C(2'p5')G(2'p5')). We show that the 2',5'-linked RNA loop adopts a conformation that is quite different from that previously observed for the native 3',5'-linked RNA loop. The 2',5'-RNA loop is stabilized by (a) U:G wobble base pairing, with both bases in the anti conformation, (b) extensive base stacking, and (c) sugar-base contacts, all of which contribute to the extra stability of this hairpin structure. PMID- 14565497 TI - Structural characterization of LNA and alpha-L-LNA hybridized to RNA. AB - LNA and alpha-L-LNA are promising candidates for the development of efficient oligonucleotide-based therapeutic agents. Here, we present a short overview of the structural results we have obtained for LNA:RNA and alpha-L-LNA:RNA hybrids. Specifically, we have shown that LNA acts as an A-type mimic, while alpha-L-LNA acts as a B-type mimic when built into oligonucleotides. PMID- 14565498 TI - Interactions of trimeric purine nucleoside phosphorylases with ground state analogues--calorimetric and fluorimetric studies. AB - Binding enthalpies, dissociation constants and stoichiometry of binding for interaction of trimeric calf spleen and Cellulomonas sp. purine nucleoside phosphorylases with their ground state analogues (substrates and inhibitors) were studied by calorimetric and spectrofluorimetric methods. Data for all ligands, with possible exception of hypoxanthine, are consistent with three identical non interacting binding sites. PMID- 14565499 TI - Crystal structure of calf spleen purine nucleoside phosphorylase in a complex with multisubstrate analogue inhibitor with 2,6-diaminopurine aglycone. AB - The crystal structure at 2.05 A resolution of calf spleen PNP complexed with stoichiometric concentration of acyclic nucleoside phosphonate inhibitor, 2,6 diamino-(S)-9-[2-(phosphonomethoxy)propyl]purine, in a new space group P2(1)2(1)2(1) which contains two full trimers in the asymmetric crystal unit is described. PMID- 14565501 TI - Influence of the length of the phosphate chain in mRNA 5' cap analogues on their interaction with eukaryotic initiation factor 4E. AB - The recognition of the 5'mRNA cap structure m7G(5')ppp(5')N by one of the components of the initiation translation machinery, the eIF4E factor, plays a pivotal role in regulation of the protein synthesis. In the present study we have shown two opposing roles of the cap phosphate chain in the specific eIF4E-cap interaction. The extension of the phosphate chain enhances the binding of the cap to the unphosphorylated eIF4E but destabilises the eIF4E-cap complex in case of the phosphorylated protein. PMID- 14565500 TI - Binding studies of eukaryotic initiation factor eIF4E with novel mRNA dinucleotide cap analogues. AB - Studies on the interaction of the murine translation initiation factor 4E with two new-synthesized cap-analogues, modified at C2' of 7-methylguanosine, have been performed by means of the fluorescence titration method. No difference in the binding affinity for eIF4E was observed compared with the "anti reversed" cap analogues, possessing the analogous modifications at C3'. Potential significance of the novel caps as research tools for examination of the nuclear cap binding complex CBC80/20 has been discussed. PMID- 14565502 TI - Interaction between yeast eukaryotic initiation factor eIF4E and mRNA 5' cap analogues differs from that for murine eIF4E. AB - Measurements of interaction of 7-methyl-GTP eIF4E from S. cerevisiae were performed by means of two methods: Isothermal Titration Calorimetry (ITC) and fluorescence titration. The equilibrium association constants (Kas) derived from the two methods show significantly different affinity of yeast eIF4E for the mRNA 5' cap than those of the murine and human proteins. The observed differences in the Kas values and the enthalpy changes of the association (deltaH(o)) suggest some dissimilarity in the mode of binding and stabilization of cap in the complexes with eIF4E from various sources. PMID- 14565503 TI - Cell surface oligonucleotide-binding proteins of human squamous carcinoma A431 cells. AB - Affinity modified with Flu-DAP-p(N)16degU oligonucleotide-binding proteins were isolated by affinity chromatography using Ultrogel A2-anti fluorescein antibodies. After separation by SDS-PAGE the proteins with molecular masses about 68 kDa were MS/MS sequenced and identified as keratin K1, keratin K10, keratin K2e and albumin. PMID- 14565504 TI - Geniom technology--the benchtop array facility. AB - Febit AG develops an integrated benchtop instrument for in situ microarrays preparation, hybridization, readout and data analysis. PMID- 14565505 TI - Resistance of human cytomegalovirus to D- and L-ribosyl benzimidazoles as a tool to identify potential targets for antiviral drugs. PMID- 14565506 TI - Linear polyethylenimine as a tool for comparative studies of antisense and short double-stranded RNA oligonucleotides. AB - Despite the recently enlarged field of available RNA knock-down technologies, e.g., antisense oligonucleotides (ASOs) and duplexes of synthetic 21 nucleotides RNAs (siRNAs), no versatile transfection reagent has been reported to deliver different nucleic acids formats at high rates of efficiency. We have evaluated the versatility and efficacy of linear PEI in transfecting and properly delivering a broad panel of nucleic acids such as short oligonucleotides and double-stranded RNA into cells in culture. PMID- 14565507 TI - The use of triisopropylsilyl-oxymethyl (TOM) in the synthesis of anti-telomerase 2-5A antisense compound RBI 011. AB - The 2-5A antisense compound RBI 011 targeting telomerase RNA was synthesized using the triisopropylsilyl-oxymethyl (TOM) group for the 3'-hydroxyl protection of 2',5'-linked RNA. PMID- 14565508 TI - The role of topotecan in treating small cell lung cancer: second-line treatment. AB - Small-cell lung cancer (SCLC) is highly chemosensitive but up to 70% of patients with limited disease and more than 90% of patients with extensive disease will relapse after first-line treatment. There are several standard chemotherapy regimens used for second-line treatment yet the prognosis for patients requiring this treatment remains poor. The topoisomerase-I inhibitor, topotecan, has achieved response rates of up to 22% in previously treated patients with SCLC and survival almost double that achieved with other single agents. Compared with cyclophosphamide/doxorubicin/vincristine (CAV), single-agent topotecan achieved a higher response rate, longer survival and statistically significant improvements in dyspnea, hoarseness, fatigue, anorexia and interference with daily activities. Brain metastases are common in SCLC. Topotecan crosses the blood-brain barrier and shows promise for the management of brain metastases. PMID- 14565509 TI - Topotecan as first-line therapy for small cell lung cancer. AB - Although current treatments for small-cell lung cancer (SCLC) yield objective response rates exceeding 50%, most patients relapse. Hence, research into the identification of novel agents and combinations that may improve therapy is ongoing. Topotecan, an established treatment for patients with recurrent SCLC, is being investigated as first-line therapy for SCLC because of its novel mechanism of action, non-cumulative toxicity and in vitro synergy with other active agents. Several phase II studies of doublet and triplet combination therapy with other agents, including paclitaxel, cisplatin, carboplatin and etoposide, have reported promising results for first-line treatment of SCLC. For example, in combination with paclitaxel, complete and overall responses were 3-67% and 45-100%, respectively, in extensive-stage disease. Furthermore, two studies of the triplet combination of topotecan with paclitaxel plus carboplatin yielded impressive complete response rates of 37-51% in limited-stage SCLC. The most frequent adverse events associated with topotecan-based regimens have been reported as neutropenia and thrombocytopenia so growth factor support is often incorporated into treatments. Several ongoing phase III studies will help to clarify the role of topotecan in the first-line treatment of SCLC. PMID- 14565510 TI - Single-agent chemotherapy for non-small cell lung cancer. AB - The survival and quality of life benefits of combination chemotherapy in patients with non-small cell lung cancer (NSCLC) are now well recognized. Since many clinical trials have been conducted in relatively young patients with good performance status, many elderly patients and patients with poor performance status are not offered chemotherapy because of concerns about higher risks of toxicity. The newer agents, including topotecan, are active as single agents in NSCLC, achieving response rates of up to 30%. Overall survival and symptoms may be improved when these agents are added to best supportive care. They are well tolerated in both elderly patients and patients with poor performance status. The major toxicity with the standard 5-day administration schedule of topotecan is myelosuppression, but weekly schedules may offer reduced toxicity while maintaining efficacy. Thus, single-agent therapy with newer agents is generally considered in elderly and poor performance status patients. However, combination chemotherapy may also be appropriate for some patients in these subgroups. Future studies of chemotherapy in NSCLC should not exclude elderly patients and patients with poor performance status. PMID- 14565511 TI - Combination chemotherapy with topotecan for non-small cell lung cancer. AB - Patients with non-small cell lung cancer (NSCLC) typically receive platinum-based combination chemotherapy. In spite of improvements in symptoms and survival, response rates remain low and newer agents are being investigated. The newer agents may offer increased efficacy and reduced toxicity compared with established agents and regimens. The topoisomerase-I inhibitor, topotecan, achieves single-agent response rates of 4-25% in NSCLC. Topotecan has also been studied in combinations: a combination of topotecan, administered using the standard 5-day schedule, with cisplatin was effective but was associated with myelosuppression. The combination of topotecan plus carboplatin may be better tolerated and warrants further investigation. Topotecan was also combined with newer agents (gemcitabine, vinorelbine, docetaxel, paclitaxel) using a range of different administration schedules of topotecan. Response rates of up to 30% were achieved. A weekly schedule of topotecan was effective and well tolerated and was also convenient for healthcare professionals and patients. PMID- 14565512 TI - Weekly topotecan in the management of lung cancer. AB - Chemotherapy agents require a range of administration schedules, including 3 weekly, 4-weekly and daily administration. Some agents, for example gemcitabine and vinorelbine, have been developed for use in a weekly regimen. The possibility of administering other agents using a weekly schedule is being investigated. Weekly schedules offer practical benefits in terms of convenience to patients and allow drugs to be combined more easily. In addition, toxicity may be reduced. The standard 5-day schedule of topotecan has demonstrated effectiveness and patient benefits. Topotecan at this dose is generally well tolerated, with dose-limiting myelosuppression. Preclinical data supported intermittent dosing with topotecan and clinical studies with weekly dosing in ovarian cancer have indicated reduced myelosuppression compared with the 5-day regimen. Several studies in non-small cell lung cancer investigated topotecan combined with cisplatin or gemcitabine and confirmed these findings. However, further studies are needed to confirm that efficacy of topotecan (response and survival) is maintained with the altered regimen. PMID- 14565513 TI - New approaches to symptom improvements in lung cancer patients. AB - The efficacy of chemotherapy in treating lung cancer continues to be assessed by the impact treatment has on disease progression and survival. Where differences between treatments are marginal and survival times cannot be realistically extended, symptom relief and improving health-related quality of life (QoL) become treatment priorities. Clinical trials in lung cancer often now include an assessment of symptom relief and QoL, and have confirmed the benefit of chemotherapy compared with best supportive care. Collection of symptom relief and QoL data is based on subjective reporting by patients, which can be influenced by extraneous and confounding factors. To ensure reproducibility and interpretability, standardized, validated instruments with known psychometric properties and culturally adapted language are required to capture QoL data from lung cancer patients in clinical trials. A number of available questionnaires, developed and validated for use in oncology studies, are reviewed in this paper. A selection of published lung cancer studies that have used these instruments are described and their outcomes summarized. These studies show that symptom assessment can be integrated in study protocols successfully, but there is a recognized need for improved and consistent measurement in future studies. PMID- 14565514 TI - Options for first- and second-line therapy in small cell lung cancer--a workshop discussion. AB - In a case study-based workshop, physicians were asked to discuss various aspects of patient management in small cell lung cancer (SCLC). For first-line chemotherapy, most investigators recommended treatment with etoposide/cisplatin, with possible dosing variations according to tolerability and convenience. In France (but not elsewhere), medical oncologists tend to use a four-drug regimen (etoposide/cisplatin/cyclophosphamide/epirubicin), based on the results of an extensive-stage SCLC trial. Alternative first-line regimens, such as vincristine/ifosfamide/carboplatin/etoposide (VICE) and topotecan/platinum, are currently being explored. Options for therapy in patients with recurrent disease are more varied, although there was consensus that active treatment at relapse should be considered. Regimens include topotecan (alone or in combination), cyclophosphamide/doxorubicin/vincristine (CAV) and re-induction with the earlier first-line agents. Studies are also investigating the potential benefits of other combinations, including topotecan/vinorelbine and paclitaxel/carboplatin. For patients with relapsed extensive-stage SCLC and brain metastases, whole brain radiation therapy was considered appropriate for both palliative and therapeutic reasons. The potential role of combination therapy with topotecan/temozolomide, both of which cross the blood-brain barrier, is currently being investigated. PMID- 14565515 TI - The 'capsule' of pituitary macroadenomas represents normal pituitary gland: a histopathological study. AB - The goal of trans-sphenoidal pituitary adenoma surgery is radical excision of the tumour with preservation of endocrine function. Our hypothesis was that, even in macroadenomas extending into the suprasellar cistern, the 'capsule' of the tumour is the compressed normal pituitary gland. The biopsy material from 126 patients with pituitary macroadenomas were retrospectively reviewed to look for the presence of normal adenohypophysis. Fourteen patients with macroadenomas operated trans-sphenoidally were studied prospectively, sampling tissue from the periphery of the tumour for histopathology. From the retrospective data, we found that normal adenohypophysis was more often found at histopathology in the extracapsular excisions, rather than in the intracapsular excisions. In the 14 patients studied prospectively, normal adenohypophysial tissue was found histologically at all sites sampled except in areas where the tumour was invasive. In conclusion, while an extracapsular excision would offer the best chance for a surgical cure, preserving parts of the capsule may preserve normal and possibly functioning gland. PMID- 14565516 TI - Meningiomas induced by high dose cranial irradiation. AB - The authors present four patients with a history of high dose cranial irradiation, who were diagnosed years later to have symptomatic meningiomas requiring surgical management. Relevant literature pertaining to these rare tumours is reviewed and their unusual characteristics highlighted. Their aetio pathogenesis and management strategies are discussed. PMID- 14565517 TI - Traumatic epidural haematoma of the posterior fossa in childhood: 16 new cases and a review of the literature. AB - A series of 16 children (nine males, seven females) with traumatic epidural haematoma of the posterior fossa (TEHPF) is presented. All patients had a history of mild to moderate trauma to the occiput. The causes were falls in 15 and traffic accident in one case. Four of the 16 cases had depressed fractures of the occipital bone. Surgical intervention was performed in all cases. In 14 patients the outcome was good, one patient had moderate disability and one died. Our study revealed that early diagnosis and urgent surgical intervention may give the patient a chance of total recovery and craniotomy with haematoma evacuation is an appropriate surgical technique as in the case of supratentorial extradural hematoma. PMID- 14565518 TI - Neurological presentation and long-term outcome following operative intervention in patients with meningocele manque. AB - We review and describe the neurological presentation and long-term outcome of patients with meningocele manque, and describe the dysraphic features associated with this entity. Our series of patients was collected over a 25-year period with a mean follow-up of 11.5 years. The mean age of presentation was 10.4 years and 72% of the patients presented with an abnormal neurological examination. At most recent follow-up, symptoms were stable, improved, and progressed in 47, 37 and 16%, respectively. Meningocele manque was associated with focal hirsutism in 37% of the cases. These bands were primarily found in the lumbar spine and involved two sequential vertebral levels in 42% of cases. Sectioning of meningocele manque has good long-term results in the majority of patients. These bands should be sought in the evaluation of patients with spinal dysraphism and surgically transected. PMID- 14565519 TI - Dysembryogenetic spinal tumours in adults without dysraphism. AB - The authors report 15 patients with spinal intradural dysembryogenetic tumours with clinical onset in adult age in the absence of clinical and radiological signs of dysraphism. The series includes seven lipomas, four epidermoid cysts, three dermoid cysts and one teratoma. The tumour site was the thoracic region in three cases, the lumbar cord and conus in six, the cauda equina in four and the filum terminale in two. Among 14 patients operated upon, the surgical removal was complete in eight cases, subtotal in two and partial in four. Recurrence was noticed only in one subtotally resected thoracic epidermoid cyst. Magnetic resonance imaging allows a precise diagnosis of these lesions, mainly of small lipomas and dermoids of the conus and filum, where a tethered conus is responsible for clinical symptoms. Dysembryogenetic spinal rumours that become symptomatic in adult age may require surgical treatment. Reduction of the mass and release of any associated tethered neural elements are the goal of surgery for spinal lipomas, whereas epidermoid and dermoid cysts require a more radical treatment. However, even partial resections to avoid neural damage result in a good clinical outcome and very low risk of recurrence. PMID- 14565520 TI - Venous ligation of dural arteriovenous fistulae. AB - This study evaluated the clinical outcomes following venous ligation for the treatment of dural arteriovenous fistulas (DAVF) with leptomeningeal or cortical venous drainage. The authors conducted a single-centre, retrospective study of 15 patients with DAVF treated consecutively between 1990 and 2000. There were eight female patients of mean age 60 years (range 36-71). Eight patients presented with haemorrhage, two with mass affect, two with headaches, one with seizures and two patients were asymptomatic. Four fistulas were located in the petrosal region, three at the torcula, three in the transverse sigmoid region, three in the parasagittal region and two at the foramen magnum. Leptomeningeal or cortical veins drained all DAVF. Eight patients underwent preoperative embolization of the arterial feeders. Venous ligation resulted in obliteration of all fistula confirmed by direct visualization of the thrombosed veins and/or radiographic follow up. Mean follow up period was 37 months with a range of 1-112 months. All symptomatic patients improved and there was no surgical morbidity. These results indicated that venous ligation to obliterate DAVF with leptomeningeal or cortical drainage is safe and effective. With appropriate neuroradiological diagnosis and preoperative neuroendovascular intervention, this technique resulted in obliteration of DAVF in 15 patients without significant morbidity. PMID- 14565521 TI - Chronic subdural haematoma treated by craniotomy, durectomy, outer membranectomy and subgaleal suction drainage. Personal experience in 39 patients. AB - According to the CT and MRI appearances, 39 chronic subdural haematoma (CSDH) patients were suspected of having solid clots and/or a high likelihood of loculation. Craniotomy was planned from the start. Beside the better exposure, excision of the dura and outer membrane, assumed to be the source of haematoma fluid, this is an additional step to minimize the incidence of significant recollection. There were no additional operative or postoperative cranial and/or systemic complications when compared with other minor procedures. Two patients (5%) required once percutaneous tapping and aspiration. Accordingly, if a case is considered to be better managed with craniotomy, durectomy and outer membranectomy this is an easy and safe technique with minimal incidence of recollection, morbidity and mortality. PMID- 14565522 TI - Symptomatic intraspinal pneumocoele resulting from closed head injury. AB - Intraspinal pneumocoele is rarely associated with closed head injury. We describe a case of intra-spinal pneumocoele following a closed head injury. We believe this to be the first case presenting with radicular symptoms. PMID- 14565524 TI - Subdural haematoma after trans-sphenoidal surgery. AB - We report a patient with pituitary adenoma who developed a subdural haematoma following trans-sphenoidal surgery and conservative management of a postoperative CSF leak with a lumbar drain. The possible causes and methods of treatment are discussed and the neurosurgeon is alerted to the possibility of its development in similar situations. PMID- 14565523 TI - Spontaneous anaplasia in pilocytic astrocytoma of cerebellum. AB - We report a cystic cerebellar astrocytoma with a mural nodule that contained an additional focus of astrocytoma with the histological features of anaplasia, and showed up to 48% of aneuploid and 3% S-phase cells on flow cytometry. This focus was detectable on the enhanced, as well as non-enhanced T1 and T2 images. This appears to be the first case of pilocytic astrocytoma of cerebellum with focal anaplasia detected on histological and radiological studies. PMID- 14565525 TI - Extradural spinal cord compression by rheumatoid nodule. AB - Rheumatoid disease is a systemic disorder affecting multiple organs. It is known to affect the nervous system in a variety of ways, but its presentation with spinal cord compression by a rheumatoid nodule is rare. We report two cases presenting with cord compression by a rheumatoid nodule who underwent surgical decompression with good recovery. PMID- 14565526 TI - Recurrent intracranial sarcoma mimicking chronic subdural haematoma. AB - Primary sarcomas of the central nervous system and their coincidence with a subdural haematoma are each rare. We describe an unusual case of unsuspected dural spreading of a recurrent spindle cell sarcoma concealed in a chronic subdural haematoma, which occurred months after microsurgical resection and external radiation of a temporal parenchymatous sarcoma. PMID- 14565527 TI - Chordoma presenting as pseudoprolactinoma. AB - Chordomas are relatively rare neoplasms that appear to originate from benign remnants of primitive notochord. They account for about 0.15% of all intracranial neoplasm and are the most common of extradural neoplasms that involve the clivus. Although they present mostly with signs and symptoms of other intracranial lesions they rarely present with high prolactin levels mimicking prolactinomas. We report a case of chordoma presenting as a pseudoprolactinoma. We discuss the presentation, radiological features and treatment of this rare presentation. PMID- 14565528 TI - Adult onset hydrocephalus and situs inversus: new autosomal dominant syndrome? AB - Familial adult onset hydrocephalus is unusual and its genetic basis is unknown. We report a two-generation family with three members having hydrocephalus, two of them with complete situs inversus. Laterality syndromes were considered in the differential diagnosis and are unlikely. We suggest that this family represents a previously unrecognised form of autosomal dominant late onset hydrocephalus with situs inversus. PMID- 14565529 TI - Sudden death secondary to delayed closure of endoscopic third ventriculostomy. AB - An 11-year-old boy who underwent an initially successful endoscopic third ventriculostomy (ETV) died 4 months later secondary to acute hydrocephalus. This is the first report of sudden death caused by delayed closure of ETV. Guidelines to patients, parents and primary caregivers should include the possibility of ETV failure and encourage early neurosurgical consultation when symptoms of raised intracranial pressure occur. PMID- 14565530 TI - Trans-sternal decompression of postlaminectomy caries of the upper dorsal spine. PMID- 14565531 TI - Bilateral sixth nerve palsy: a rare presentation of postoperative posterior fossa extradural haematoma. PMID- 14565532 TI - Low-grade astrocytoma of the spinal cord associated with neurofibromatosis type 1: report of a case with poor correlation between histopathology and prognosis. PMID- 14565533 TI - Endovascular treatment of anterior communicating artery aneurysms: results of clinical and radiological outcome in Newcastle. AB - The results of endovascular management of anterior communicating artery aneurysms (ACoAA) using Guglielmi Detachable Coils (GDC) are presented. We detail the clinical and radiological features, and postoperative clinical and radiological outcome in a consecutive series of patients. We have prospectively collected comprehensive data on our patients with SAH since 1989. This study reports on patients admitted between January 1990 and December 1998, and focuses on 30 patients who had their ACoAA endovascularly treated. An independent observer (TE) carried out long-term follow-up in January 2002. Statistical analysis was performed to study the relationship between clinical factors, the radiological morphology of aneurysms and the long-term outcome. The age ranged from 25 to 74 years (median: 54) and endovascularly treated ACoAA were more common in women, 19 (63%) compared with men, 11 (37%). Seventy-seven per cent were in good grade (WFNS 1 & 2) before treatment. Three patients rebled before treatment. The aneurysms were less than 10 mm in maximal diameter in 27/30 patients. The follow up period was from 1 to 53 months (mean 32.5, median 36.6 months). Excellent outcome was recorded for 11 patients (36.7%), good in seven patients (23.3%), fair in three patients (10%) and poor in four cases (13.3%). Five patients had died (mortality 16.7%). Favourable outcome was achieved in younger patients, women, and in those who presented in grades 1 and 2. The long-term radiological follow-up revealed residual necks in 13 patients. Only one has required retreatment and no rebleed has occurred in 3-6 years. This study reports a contemporary experience with the endovascular management of ACoAA. Long-term stability of the coil and good outcome is demonstrated. Endovascular treatment using GDC is an efficient technique for treating acutely ruptured ACoAA with little additional morbidity. The clinical and radiological results are comparable with those in the literature. PMID- 14565534 TI - Autonomic innervation of immune organs and neuroimmune modulation. AB - 1. Increasing evidence indicates the occurrence of functional interconnections between immune and nervous systems, although data available on the mechanisms of this bi-directional cross-talking are frequently incomplete and not always focussed on their relevance for neuroimmune modulation. 2. Primary (bone marrow and thymus) and secondary (spleen and lymph nodes) lymphoid organs are supplied with an autonomic (mainly sympathetic) efferent innervation and with an afferent sensory innervation. Anatomical studies have revealed origin, pattern of distribution and targets of nerve fibre populations supplying lymphoid organs. 3. Classic (catecholamines and acetylcholine) and peptide transmitters of neural and non-neural origin are released in the lymphoid microenvironment and contribute to neuroimmune modulation. Neuropeptide Y, substance P, calcitonin gene-related peptide, and vasoactive intestinal peptide represent the neuropeptides most involved in neuroimmune modulation. 4. Immune cells and immune organs express specific receptors for (neuro)transmitters. These receptors have been shown to respond in vivo and/or in vitro to the neural substances and their manipulation can alter immune responses. Changes in immune function can also influence the distribution of nerves and the expression of neural receptors in lymphoid organs. 5. Data on different populations of nerve fibres supplying immune organs and their role in providing a link between nervous and immune systems are reviewed. Anatomical connections between nervous and immune systems represent the structural support of the complex network of immune responses. A detailed knowledge of interactions between nervous and immune systems may represent an important basis for the development of strategies for treating pathologies in which altered neuroimmune cross-talking may be involved. PMID- 14565535 TI - Inhibition of calcium/calmodulin-dependent protein kinase II normalizes diabetes induced abnormal vascular reactivity in the rat perfused mesenteric vascular bed. AB - 1. Calcium/calmodulin-dependent protein kinase II (CaMKII) has an important function in mediating insulin release but its role in the development of diabetes induced cardiovascular complications is not known. 2. We investigated the ability of a chronic administration of KN-93 (5 mg kg(-1) alt diem for 4 weeks), an inhibitor of CaMKII, to modulate the altered vasoreactivity of the perfused mesenteric bed to common vasoconstrictors and vasodilators in streptozotocin (STZ)-induced diabetes. 3. The vasoconstrictor responses induced by noradrenaline (NE), endothelin-1 (ET-1), and angiotensin II (Ang II), were significantly increased whereas, vasodilator responses to carbachol and histamine were significantly reduced in the perfused mesenteric bed of the STZ-diabetic rats as compared with non-diabetic controls. 4. Inhibition of CaMKII by KN-93 treatment did not affect blood glucose levels but produced a significant normalization of the altered agonist-induced vasoconstrictor and vasodilator responses. KN-93 did not affect agonist-induced responses in control animals. In addition, KN-93 significantly reduced weight loss in diabetic rats. 5. The present data suggest that CaMKII is an essential mediator in the development of diabetic vascular dysfunction and may also play an important role in signalling pathways leading to weight loss during diabetes. PMID- 14565536 TI - Vasopressin-induced facilitation of adrenergic responses in the rat mesenteric artery is V1-receptor dependent. AB - 1. The present study was designed to analyse the possible involvement of V1- and V2-receptors in vasopressin (AVP)-induced facilitation of the sympathetic nervous system. Furthermore, we aimed to determine whether the site of facilitation by AVP is located pre- or postsynaptically. 2. Electrical field stimulation (EFS) was applied on the rat mesteric artery to activate the sympathetic nervous system. In addition, we evaluated the direct vascular effects of AVP. The postsynaptic effect of AVP on the sympathetic nervous system was investigated by exposing the vessels to exogenous noradrenaline. These experiments were performed in the absence or presence of selective V1 and V2 receptor antagonists SR 49059 and SR 121463, respectively. Desmopressin was applied as a selective V2 agonist. 3. The direct vasoconstrictor effect of AVP was antagonized by SR 49059 and not by SR 121463. Desmopressin neither showed any direct vasoconstrictor effect nor produced vasodilatation after a precontraction induced by noradrenaline (10 microM). The EFS-induced rise in vascular tone could be increased by a sub pressor concentration of AVP. This fascilitation could be antagonized by SR 49059, but not by SR 121463. Desmopressin did not influence the increase in vascular tone during EFS. Vasoconstriction induced by exogenous noradrenaline could be facilitated by a sub-pressor concentration of AVP and this selective postsynaptic effect could be antagonized by V1-receptor blockade. 4. In conclusion, the AVP-induced facilitation of the sympathetic nervous system is completely V1-receptor dependent and at least partly postsynaptically mediated. PMID- 14565537 TI - Responsiveness, affinity constants and beta 2-adrenoceptor reserves for isoprenaline on portal veins from normo- and pre- and hypertensive rats. AB - 1. This study used contractility methods with the portal veins of 5- and 14-week old Wistar-Kyoto normotensive rats (WKY) and spontaneously hypertensive rats (SHRs). The SHRs are prehypertensive at 5 weeks. 2. The first part of our study was to determine whether the responsiveness to isoprenaline and forskolin was altered in the maturation of portal veins from normo- and prehypertensive rats. The responses to forskolin were similar on the portal veins of 5- and 14-week-old WKY and SHRs. 3. The sensitivity and maximum responses to isoprenaline were similar on portal veins of 5- and 14-week-old WKY. The sensitivity and maximum responses to isoprenaline were lower on the portal veins of 5-week-old SHRs (pD2 = 8.25, maximum = 85%) than age-matched WKY (pD2 = 8.79, maximum = 96%); these differences are not caused by hypertension. At 14 weeks, the sensitivity was similar (WKY pD2 = 8.74, SHR pD2 = 8.65) but the maximum responses to isoprenaline were lower on the portal veins SHRs (77%) than WKY (97%). Thus, the sensitivity to isoprenaline increases with the development of hypertension in the SHR portal vein. 4. The second part of the study was to determine whether the affinity for isoprenaline at beta2-adrenoceptors and the fractional beta2 adrenoceptor occupancy-response relationships on the portal vein were altered in maturation from normo- and pre-hypertensive rats. The effects of bromoacetylalprenololmenthane (BAAM), an irreversible beta-adrenoceptor blocker, on the isoprenaline responses of 5- and 14-week-old WKY and SHRs were studied. Maturation of the WKY portal vein between 5 and 14 weeks was associated with a loss of affinity for isoprenaline (from pKA of 7.13 to 7.87), and increase in beta2-adrenoceptor reserve (from 72 to 92% at the 95% response). There were similar affinity and reserve findings in the maturation of the SHR portal vein. Thus, there are major changes in beta2-adrenoceptor structure and reserve in maturation on the portal vein that are irrespective of the development of hypertension. PMID- 14565538 TI - Rilmenidine produces mydriasis in cats by stimulation of CNS alpha 2 adrenoceptors. AB - 1. Experiments were undertaken to determine if the imidazoline/alpha2 adrenoceptor agonist, rilmenidine, would produce mydriasis in cats and, if so, to delineate its site of action and determine if this effect is mediated by imidazoline receptors or alpha2-adrenoceptors. 2. Rilmenidine produced dose related pupillary dilator responses in pentobarbital anaesthetized cats that were independent of sympathetic innervation to the iris but were dependent upon intact parasympathetic neuronal tone. The ED50 for rilmenidine-induced pupillary dilation was approximately 200 microg kg(-1), i.v., and was sustained for at least 1 h. 3. The highly selective alpha2-adrenoceptor antagonist, RS-79948, administered either before or after rilmenidine, antagonized rilmenidine-induced mydriasis. Neuronally induced reflex inhibition of parasympathetic nerve activity was also inhibited by administration of RS-79948. 4. These results suggest that rilmenidine acts like clonidine to produce pupillary dilation by inhibition of parasympathetic tone to the iris sphincter and that this central nervous system parasympatho-inhibition is mediated by alpha2-adrenoceptors, rather than imidazoline receptors. PMID- 14565539 TI - Actions of domperidone on the opioid system in isolated guinea-pig ileum: differences between dopamine antagonists on opioid system. AB - 1. To validate the relationship between the dopaminergic, opioidergic and cholinergic nervous systems, we evaluated the effect of domperidone, a dopamine (D2) antagonist, on the opioid system in myenteric plexus-longitudinal muscle preparation isolated from the guinea-pig ileum. 2. One micromolar of domperidone did not affect the 0.1 Hz-evoked (duration 0.5 ms, maximum intensity) twitch response, but concentration dependently inhibited the twitch response between concentrations of 2 and 20 microM, and the inhibition was maximum after 20-30 min at the highest concentration used (20 microM). 3. Acetylcholine-evoked contraction on basal tension was also not inhibited by 1 microM domperidone, but the contraction was concentration dependently inhibited at concentrations of 10 100 microM in a non-competitive manner. 4. One micromolar of domperidone, however, increased post-tetanic twitch inhibition, an indicator of the release of endogenous opioids. This increase was completely antagonized by 1 microM naloxone. Twitch inhibition induced by dynorphin 1-13 (0.1-10 nM) was not affected by 1 microM domperidone, but increased the maximum twitch inhibition caused by morphine (0.1-1 microM). 6. These results might reflect the existence of an interaction between the dopaminergic and opioidergic system without the inhibition of the cholinergic system. Dopamine antagonists increased opioid action, an action that may depend more on the increased release of endogenous opioids than on supersensitivity of the opioid receptor. PMID- 14565540 TI - Reflex bradycardia induced by hydralazine in sino-aortic deafferented conscious rats. AB - 1. It is generally recognized that the vasodilator hydralazine produces hypotension accompanied by baroreflex-mediated tachycardia. In some experimental conditions, however, the accompanying heart rate change is bradycardia, a paradoxical response which has not been satisfactorily explained. The present study examined the possibility of hydralazine-induced bradycardia being mediated by vagal or sympathetic afferents activated by changes in left ventricular pressure. 2. Systolic blood pressure and heart rate responses to hydralazine were recorded in conscious normotensive intact rats by a tail cuff method and compared with responses in animals subjected to previous sino-aortic deafferentation (SAD) to remove the influence of the arterial baroreflex. Responses were also obtained after blockade of myocardial afferent vagal C-fibres with urethane, of efferent vagal impulses to the heart with methylatropine, of positive inotropic effects of hydralazine with atenolol, and of prostanoid sensitization of myocardial nerve fibres with indomethacin. 3. Hydralazine produced hypotension and tachycardia in intact rats, and hypotension and bradycardia in SAD animals. In intact rats, this pattern was not affected by any of the pretreatments, while in SAD rats, all pretreatments reversed the bradycardia to hydralazine. 4. The present results indicate that suppression of the arterial baroreflex by SAD propitiates the appearance of a bradycardiac response to hydralazine. This reaction probably results from activation of a vagal cardiodepressant reflex originating in the heart, as suggested by its blockade by drugs acting at various sites along the reflex arch. PMID- 14565541 TI - The effect of chronic ethanol administration on nitric oxide-mediated responses in rat isolated trachea preparation. AB - 1. In the present study, we investigated the effect of chronic ethanol administration on nitric oxide (NO)-mediated responses in rat isolated trachea preparation. 2. Ethanol was given to rats in a modified liquid diet for 21 days. Isolated tracheal rings were then used to obtain responses to electrical field stimulation (EFS) after precontraction with 100 microM histamine. The parameters of field stimulation were as follows: supramaximal voltage of 50 V, 0.5 ms duration, 10-s train; 0.5, 1, 3, 5, 10, 20, 30 and 50 Hz at 2-min intervals. The effects of L-and D-arginine (10(-6) M) on the responses to field stimulation (10 20 Hz) were studied. In other experiments, we tested the effects of N(omega) nitro-L-arginine methyl ester (L-NAME, 10(-6)-10(-5) M) and SIN-1 (10(-6)-10(-5) M) on the responses to field stimulation. 3. Electrical field stimulation induced relaxation responses in the tracheal rings precontracted with histamine from control- and ethanol-treated rats. The relaxation responses induced by EFS were significantly reduced in the tracheal rings precontracted with histamine from ethanol dependent group. The responses induced by EFS in both groups were completely abolished by tetrodotoxin (1 microM), but unaffected by hexamethonium (1 microM). Incubation with D-arginine did not cause statistically significant increases in relaxation responses to EFS in both groups. L-Arginine (10(-6) M) caused statistically significant increases in relaxation responses to EFS in control rats, but not in ethanol dependent rats. Incubation with L-NAME (10(-6) 10(-5) M) caused statistically significant inhibition of the relaxation responses to EFS in both groups. SIN-1 (10(-6)-10(-5) M) induced significantly increase in relaxation responses to EFS in both groups. 4. Our results suggest that the possible mechanism responsible for inhibition of tracheal inhibitory responses to EFS in ethanol-dependent rats may be a reduction in production of NO and in the uptake of L-arginine. PMID- 14565543 TI - Determination of caffeine using oscillating chemical reaction in a CSTR. AB - A new analytical method for the determination of caffeine by the sequential perturbation caused by different amounts of caffeine on the oscillating chemical system involving the manganese(II)-catalyzed reaction between potassium bromate and tyrosine in acidic medium in a CSTR was proposed. The method exposed for the first time in this work. It relies on the relationship between the changes in the oscillation amplitude of the chemical system and the concentration of caffeine. The calibration curve fits a second-order polynomial equation very well when the concentration of caffeine over the range 4.0 x 10(-6) - 1.2 x 10(-4) M (r = 0.9968). The effect of influential variables, such as the concentration of reaction components, injection point, temperature, flow rate and stirring rate were studied. Some aspects of the potential mechanism of action of caffeine on the chemical oscillating system were also discussed. A real sample was determined and the result was satisfactory. PMID- 14565542 TI - Increased blocking activity of combined tachykinin NK1- and NK2-receptor antagonists on tachykinergic bronchomotor responses in the guinea-pig. AB - 1. The present study compared the effect of the administration of tachykinin NK1- and NK2-receptor antagonists alone and in combination on exogenous and endogenous tachykinin-induced contractions using three different guinea-pig airway preparations: isolated bronchus, isolated perfused lung and in vivo. 2. In the isolated bronchi, the tachykinin NK1-receptor antagonist CP 99994 (0.01-1 microM) produced concentration-dependent inhibition of contractions induced the tachykinin NK1-receptor agonists substance P (SP) and [Met-OMe11] SP ([Met OMe11]SP), whereas the tachykinin NK2-receptor antagonist SR 48968 (0.1 microM) had no effect. SR 48968 (0.001-0.01 microM) concentration-dependently inhibited contractions induced by the tachykinin NK2-receptor agonists neurokinin A (NKA) and [beta-Ala8]-neurokinin A (4-10) ([betaAla8]-NKA) whereas CP 99994 (0.1 microM) did not inhibit the contractions. The contractile activity of capsaicin, an agent that releases endogenous tachykinins from sensory C-fibres, was inhibited in a concentration dependent manner by SR 48968 (0.001-0.03 microM) but not by CP 99994 (0.1 microM). Combination of CP 99994 and SR 48968 caused increased inhibitory effects on the concentration-response curves to SP, [Met OMe1l]SP, NKA, [beta-Ala8]-NKA and capsaicin. 3. In isolated perfused lungs, SR 48968 concentration (0.01-10 microM) dependently inhibited NKA-, [beta-Ala8]-NKA- and capsaicin-induced bronchoconstriction whereas CP 99994 (30 microM) had no effect on SP-, NKA-, [beta-Ala8]-NKA- and capsaicin-induced bronchoconstriction. Combination of inactive concentrations of CP 99994 and SR 48968 produced an increased inhibitory effect on all previous stimuli-induced bronchoconstriction. 4. In in vivo guinea-pig studies, intravenous and oral pretreatment with SR 48968 (0.01-1 mg kg(-1) i.v. and 0.1-3 mg kg(-1) p.o., respectively), but not with CP 99994 (1 mg kg(-1) i.v. and 0.3-30 mg kg(-1) p.o., respectively), produced a dose dependent inhibition of the bronchoconstrictor responses induced by NKA, [beta Ala8]-NKA and capsaicin. CP 99994 intravenously (0.3 mg kg(-1)) and orally (3-10 mg kg(-1)) inhibited SP-induced bronchoconstriction only. Intravenous and oral low dose combinations of CP 99994 and SR 48968 produced an increased inhibition of SP-, NKA-, [beta-Ala8]-NKA- and capsaicin-induced bronchoconstriction, respectively. The present data indicate that combined tachykinin NK1- and NK2 receptor antagonist treatment compared with single antagonist treatment, using CP 99994 and SR 48968, produced an augmented blockade of tachykinin NK1-, NK2- and capsaicin-mediated contractions in guinea pig airways. These findings support the hypothesis that a dual NK1- and NK2-receptor antagonist may provide an advantage over single activity tachykinin NK1- or NK2-receptor antagonists in pulmonary obstructive diseases. PMID- 14565544 TI - Modeling the effect of analyte and reference bandwidths on signal and noise magnitudes in spectrophotometric assays. AB - In spectrophotometric assays, it has been well established that the recorded absorption, and therefore the experimentally determined extinction coefficient, decreases as a function of detected bandwidth. This manuscript presents an expression for the extinction coefficient as a function of the critical parameter detected bandwidth per transition linewidth. Calculations for both single channel and multichannel photodetection are presented; the derived expressions are shown to be in good agreement with experimental results. It is important to realize that this systematic bias is present in dilute solutions of low absorptivity, and the experimentally recorded extinction coefficient for a molecular standard such as caffeine can vary approximately 4% or more, depending upon choice of research instrumentation. The magnitude of this bias may be sufficient to effect method robustness, cause interlaboratory discrepancies, and fail system suitability requirements for spectrophotometric assays. The signal to noise ratio, for example as analyzed in HPLC/UV-VIS detected chromatograms, is also a function of the detected bandwidths of both the analyte and reference channels. It is shown here that use of a reference can only increase the baseline noise. PMID- 14565545 TI - Study of stability of methotrexate in acidic solution spectrofluorimetric determination of methotrexate in pharmaceutical preparations through acid catalyzed degradation reaction. AB - Study of the degradation reaction of methotrexate (MTX) in acidic solution was carried out. Optimization of the experimental parameters of MTX acid hydrolysis was investigated. Spectrofluorimetric method for determination of MTX through measurement of its acid-degradation product, 4-amino-4-deoxy-10-methylpteroic acid (AMP), was developed. Stability of the standard solution of MTX prepared in sulfuric acid was discussed in the view of accelerated stability analysis. Two other comparative spectroflourimetric methods based on measuring the fluorescence intensities from either a condensation reaction with acetylacetone-formaldehyde (Hantzsch reaction) or a reaction with fluorescamine were also described. Beer's law validation, accuracy, precision, limits of detection, limits of quantification, and other aspects of analytical merit are presented in the text. The proposed methods were successfully applied for the analysis of MTX in pure drug and tablets dosage form. The sensitivity of the developed methods was favorable, so it was possible to be adopted for determination of MTX in plasma samples for routine use in high-dose MTX therapy. PMID- 14565546 TI - Determination of chromium and selected elements in multimineral and multivitamin preparations and in pharmaceutical raw material. AB - The content of elements in pharmaceutical preparations is one of the indispensable factors of the evaluation of their quality. In the present work, the following macro- and microelements Ca, Cr, Cu, Fe, Mg, Mn, Mo, P, Se and Zn were determined in multimineral and multivitamin preparations and in pharmaceutical raw material. Inductively coupled plasma mass spectrometry (ICP MS) and electrothermal atomic absorption spectrometry (ET AAS) were used throughout the study. The examined samples were dissolved in a high-pressure microwave system using concentrated nitric acid. The effect of the carbon residue in the digest solution on the determination result was eliminated by introducing an equation correcting the ArC+ interference with 52Cr. PMID- 14565547 TI - Conventional and micellar liquid chromatography method development for danazol and validation in capsules. AB - Two isocratic liquid chromatographic methods (conventional and micellar) for the determination of danazol (DZ) in capsules using canrenone (CAN) as internal standard have been developed and validated. In conventional liquid chromatography a mobile phase 35% water:acetonitrile 65%, v:v, a flow-rate 1 ml min(-1) and a C18 Hypersil ODS (250 x 4.6 mm, 5 microm) column (25 degrees C) were used. In micellar liquid chromatography (MLC) the conditions were: mobile phase 40 mM sodium dodecyl sulfate:2% pentanol, flow-rate 0.5 ml min(-1) and C18 Hypersil ODS (150 x 3.0 mm, 5 microm) column (60 degrees C). For both methods. UV absorbance detection at 280 nm was used and a separation up to base line was achieved. Prior to HPLC analysis a simple sample preparation was required. The recoveries found in the accuracy test were 99 +/- 10 and 101 +/- 8%, in conventional liquid chromatography (CLC) and MLC, respectively. Repeatability and intermediate precision expressed as R.S.D. were lower than 5% for both methods. Detection limits obtained were 2.4 and 3.0 ng g(-1) in CLC and CLM, respectively. PMID- 14565548 TI - A validated high performance liquid chromatographic method for analysis of nicotine in pure form and from formulations. AB - A reverse phase HPLC method using C18 column has been developed for the quantitative estimation of nicotine in the bulk material and formulations (extended release and immediate release dosage forms). The method is specific to nicotine (RT approximately 4.64 min, asymmetry approximately 1.75), and can resolve analyte peak from excipient interferences. It is linear (coefficient of variation approximately 0.9993), accurate (average recovery approximately 100%), and passed all the system suitability requirements. Applicability of the method was evaluated in analysis of drug-excipient compatibility samples, commercial dosage form (such as nicotine gum) and two novel in-house formulations. PMID- 14565549 TI - Quantification of 5-HMF and dextrose in commercial aqueous dextrose solutions. AB - 5-Hydroxymethylfurfural (5-HMF) is the major thermal decomposition product formed in aqueous dextrose solutions during sterilization and upon storage. Current compendial tests employ a spectrophotometric assay for 5-HMF and a separate assay based on optical rotation for dextrose. Efforts herein focused on identification of approaches to simultaneously quantify both 5-HMF and dextrose in aqueous solutions. Initial studies employed anion exchange chromatography with pulsed amperometric detection (PAD). These studies showed that the low anion exchange column loading capacity and concentration mismatch of dextrose and 5-HMF in typical commercial solutions makes simultaneous quantitation impractical at a single sample concentration. However, mixed modes of detection for each analyte using refractive index (RI) detection for dextrose and UV detection for 5-HMF proved viable for simultaneous quantitation. The performance of both approaches was evaluated and extremely high sensitivity for 5-HMF demonstrated (30-50 ppb). The formation of 5-HMF upon the forced thermal decomposition of dextrose was monitored and compared with results from the USP spectrophotometric assay. PMID- 14565550 TI - Impurity profile study of repaglinide. AB - Three unknown impurities and a byproduct in repaglinide bulk drug at levels below 0.1% (ranging from 0.05 to 0.1%) were detected by a simple isocratic reversed phase high performance liquid chromatography (HPLC) method. These impurities were isolated from crude sample of repaglinide using reversed-phase preparative high performance liquid chromatography. Based on the spectroscopic data (IR, NMR and MS) the structures of these impurities (I, II and IV) and byproduct (III) were characterised as 4-carboxymethyl-2-ethoxy-benzoic acid (I), 4 cyclohexylaminocarbamoylmethyl-2-ethoxy-benzoic acid (II), 1-cyclohexyl-3-[3 methyl-1-(2-piperidin-1-yl-phenyl)-butyl]-urea (IV) and 1,3-dicyclohexyl urea (III), respectively. Their synthesis and formation is discussed. PMID- 14565551 TI - Simple and rapid analysis of endocrine disruptors in liquid medicines and intravenous injection solutions by automated in-tube solid-phase microextraction/high performance liquid chromatography. AB - A simple and rapid method was developed for analyzing contamination of endocrine disruptors in liquid medicines and intravenous injection solutions. Endocrine disrupting compounds such as bisphenol A (BPA), alkylphenols and phthalates were quantitated by on-line in-tube solid-phase microextraction coupled with high performance liquid chromatography (in-tube SPME/HPLC) with UV detection. The liquid medicines and intravenous injection solutions could be used directly without any pretreatment, and the BPA, alkylphenols and phthalates in these solutions were automatically analyzed. The limits of quantification for these compounds were 1-10 ng/ml. Recoveries of these compounds spiked to the intravenous injection solutions was over 80%, except for some phthalates. Di-n butyl phthalate (DBP) was detected at a concentration of 7-60 ng/ml in most intravenous injection solutions in plastic containers, but it was not detected in solutions in glass bottles. Diethyl phthalate, di-n-propyl phthalate, DBP and di 2-ethylhexyl phthalate (DEHP) were also detected in syrup, lotion and eye drops in plastic containers. On the other hand, BPA and alkylphenols were not detected at all in these solutions. DEHP contamination from an administration set increased when total vitamin formulation was added to the infusion solution. DEHP was easily leached from polyvinyl chloride tubing by polysorbate 80. The in-tube SPME/HPLC method is simple, rapid and automatic, and it provides a useful tool for the screening and determination of endocrine disruptor contamination in liquid medicines and intravenous injection solutions. PMID- 14565552 TI - MECC determination of oleanolic acid and ursolic acid isomers in Ligustrum lucidum Ait. AB - In this paper, a novel method to separate and determine oleanolic acid and ursolic acid isomers in Ligustrum lucidum Ait was studied by micellar electrokinetic capillary chromatography (MECC). The baseline separation of the two analytes were obtained on the condition that the buffer contained 15 mmol/l disodium hydrogen phosphate, 15 mmol/l disodium tetreborate, 10 mmol/l SDS and 5% (v/v) alcohol. The contents of the oleanolic acid and ursolic acid were determined in L. lucidum Ait, Folium photiniae and Flos campsis; they were 78.3 (R.S.D. = 2.75%) and 20.7 mg/g (R.S.D. = 2.97%), 27.9 (R.S.D. = 3.67%) and 79.8 mg/g (R.S.D. = 3.44%), 65.5 (R.S.D. = 3.73%) and 60.4 mg/g (R.S.D. = 4.06%) (n = 5), respectively. The recoveries of the analytes in the extract of L. lucidum Ait were 102%, (R.S.D. = 2.85%) for oleanolic acid and 104% (R.S.D. = 3.21%) for ursolic acid (n = 5). With the emphasis on the effects of SDS and alcohol concentrations on the separation of the isomers were investigated. PMID- 14565553 TI - Development and optimization of a reversed-phase high-performance liquid chromatographic method for the determination of acetaminophen and its major metabolites in rabbit plasma and urine after a toxic dose. AB - A reversed-phase high-performance liquid chromatographic method with detection at 242 nm was developed, optimized and validated for the determination of acetaminophen (A) and its major metabolites glucuronide (AG) and sulfate (AS) conjugates in rabbit plasma and urine after a toxic dose. m-Aminophenol was used as internal standard (IS). A Hypersil BDS RP-C18 column (250 x 4.6 mm), 5 microm particle size, was equilibrated with a mobile phase composed of aqueous buffer solution of KH2PO4 0.05 M containing 1% CH3COOH (pH 6.5) and methanol (95:5, v/v). Its flow rate was 1.5 ml/min. Calibration curves of A, AG and AS were linear in the concentration ranges of 0.5-250, 1-200, 0.5-100 microg/ml in plasma and 1-200, 0.5-150, 0.5-100 microg/ml in urine matrix, respectively. Limits of detection and quantitation were calculated in all cases and extensive recovery studies were also performed. Intra-day relative standard deviation (R.S.D.) for A, AG and AS in plasma was less than 5, 4, 2% and in urine less than 4, 7, 4%, respectively, while the corresponding inter-day values were 7, 6, 4% and 5, 8, 6%, respectively. PMID- 14565554 TI - Estimation of uncertainty in kinetic-colorimetric assay of bacterial endotoxins. AB - The relative standard deviation (R.S.D.) of measurements is estimated in the kinetic-colorimetric assay of bacterial endotoxins without recourse to the usual repeated experiments. The measurements are the slopes of kinetic curves and two major factors are considered to cause the uncertainty of the measurements: (1) the pipetting of the sample and color development reagent; and (2) noise in the detection unit. The measurement R.S.D. is formulated as a function of endotoxin concentration. Two parameters (S.D. of the pipetted volumes and S.D. of the detector noise) are also required in the uncertainty equation, but no arbitrary coefficients are included. Since the S.D. values for pipettes and detector noise can be determined independently of the endotoxin assays, the measurement R.S.D. can be estimated by the above equation without repeating the assays. However, the calibration curve is necessary. The theoretical estimation is shown to be in good agreement with the experimental R.S.D. (n = 12) over a wide concentration range. PMID- 14565555 TI - Study of the interaction of 6-mercaptopurine with protein by microdialysis coupled with LC and electrochemical detection based on functionalized multi-wall carbon nanotubes modified electrode. AB - Microdialysis sampling coupled with liquid chromatography and electrochemical detection (LC-ECD) was developed and applied to study the interaction of 6 Mercaptopurine (6-MP) with bovine serum albumin (BSA). In the LC-ECD, the multi wall carbon nanotubes fuctionalized with carboxylic groups modified electrode (MWNT-COOH CME) was used as the working electrode for the determination of 6-MP. The results indicated that this chemically modified electrode (CME) exhibited efficiently electrocatalytic oxidation for 6-MP with relatively high sensitivity, stability and long-life. The peak currents of 6-MP were linear to its concentrations ranging from 4.0 x 10(-7) to 1.0 x 10(-4) mol l(-1) with the calculated detection limit (S/N = 3) of 2.0 x 10(-7) mol l(-1). The method had been successfully applied to assess the association constant (K) and the number of the binding sites (n) on a BSA molecular, which calculated by Scatchard equation, were 3.97 x 10(3) mol(-1) l and 1.51, respectively. This method provided a fast, sensible and simple technique for the study of drug-protein interactions. PMID- 14565556 TI - Shift in pH of biological fluids during storage and processing: effect on bioanalysis. AB - The pH of ex vivo plasma, bile and urine was monitored at different times and temperatures of storage, and following different sample processing methods such as ultrafiltration, centrifugation, precipitation and evaporation. The results showed that the pH of ex vivo plasma, bile and urine increased upon storage, and following sample processing and could lead to significant degradation of pH labile compounds. Several compounds were used to illustrate the impact of pH shifts on drug stability and interpretation of results obtained from in vivo studies. For example, after 1 h of incubation (37 degrees C) in rat plasma (pH 8.3), about 60%, of I was lost. However, in phosphate buffer, losses were about 12% at pH 7.4 and 40% at pH 8.0. Plasma pH also increased during ultrafiltration, centrifugation and extraction. After methanol precipitation of plasma proteins, and evaporation of the supernatant and redissolution of the residue, the resulting solution had a pH of 9.5. Under these conditions, II was degraded by 60% but was stable when phosphate buffer was used to maintain the pH at 7.4. The shift in plasma pH can yield misleading results from in vivo studies if the pH is not controlled. For example, the major circulating metabolite of II was also formed in plasma ex-vivo. This ex vivo degradation was prevented when blood samples were collected into tubes containing 0.1 volume of phosphate buffer (0.3 M, pH 5). The pH of ex vivo plasma can best be stabilized at physiological conditions using 10% CO2 atmosphere in a CO2 incubator. Changes in pH of ex vivo urine and bile samples can have similar adverse effect on pH-labile samples. Thus, processing of plasma samples under a 10% CO2 atmosphere is a method of choice for stability or protein binding studies in plasma, whereas citrate or phosphate buffers are suitable for stabilizing pH in bile and urine and for plasma samples requiring extensive preparations or long term storage. PMID- 14565557 TI - Development of an animal-human antibody complex for use as a control in ELISA. AB - In order to provide the equivalent of a human anti-human protein antibody as positive control in ELISAs, a goat-human antibody complex was created using chemical cross-linking. The resulting hybrid complex had a larger molecular size on HPLC and SDS-PAGE. In ELISA, the goat-human complex bound to human antigen and was detectable by a secondary anti-human conjugate. The method to make the hybrid complex is simple, cost-effective and can be used to make human-like antibodies to many human proteins. PMID- 14565558 TI - Accelerated metabolite identification by "extraction-NMR". AB - Examples of the use of extraction-NMR, an efficient and rapid method to obtain structural information on metabolites without prior separation, are described. Crude ethyl acetate extracts of in vitro microsomal incubations were analysed by NMR spectroscopy. The region downfield of 5.5 ppm in the proton spectra of these crude extracts was found to be essentially clear of endogenous resonances. As a consequence, sites of aromatic hydroxylation can often be determined without prior separation of the crude extracts. Sites of metabolism close to the aromatic region may also be accessible via two-dimensional (2D) homonuclear experiments (e.g. COSY, NOESY, TOCSY). One-dimensional (1D) and 2D fluorine experiments also can provide additional information on the structure of metabolites. Depending on the complexity of the aromatic region of the parent compound, signal overlap and the relative abundance of the individual components, extraction-NMR has the potential to provide information for unambiguous structure elucidation of two or three major metabolites. Should extraction-NMR produce inconclusive results, i.e. too many metabolites are present or metabolism occurred exclusively on aliphatic regions, it is possible to re-use the extraction-NMR sample and proceed with traditional methods of analysis. PMID- 14565559 TI - Simultaneous spectrophotometric determination of cyproterone acetate and estradiol valerate in pharmaceutical preparations by ratio spectra derivative and chemometric methods. AB - Ratio spectra derivative spectrophotometry and two chemometric methods (classical least squares, CLS and inverse least squares, ILS, were proposed for the simultaneous quantitative analysis of a binary mixture consists of cyproterone acetate (CA) and estradiol valerate (EV) in the commercial pharmaceutical preparations. In the ratio spectra derivative method, linear regression equations for both drugs were obtained by measuring the analytical signals at the wavelenghts corresponding to either maximums and minimums in the first derivative spectra of the ratio spectra. In the chemometric techniques, the concentration matrix was prepared by using the synthetic mixtures containing these drugs. The absorbance matrix corresponding to the concentration matrix was obtained by measuring the absorbances at 14 wavelengths in the range 220-290 nm for the zero order spectra. Two chemometric calibrations were constructed by using the absorbance matrix and concentration matrix for the prediction of the unknown concentrations of CA and EV in their mixture. The numerical values were calculated by using 'MAPLE V' software. The accuracy and the precision of the methods have been determined and they have been validated by analyzing synthetic mixtures containing these two drugs. The proposed methods were successfully applied to a pharmaceutical formulation, sugar-coated tablet, and the results were compared with each other. PMID- 14565560 TI - Phloroglucinol: novel synthesis and role of the magnesium cation on its binding with human serum albumin (HSA) using a biochromatographic approach based on Langmuir isotherms. AB - In this paper, a new and efficient method for synthesis of phloroglucinol with an overall yield of 60% was described. As well, the phloroglucinol association on an immobilized human serum albumin (HSA) column was analyzed in biochromatography by the determination of its Langmuir distribution isotherms. The role of the magnesium cation Mg2+ on the phloroglucinol-HSA binding process was as well analyzed. The results showed that in the Mg2+ concentration range (0.7-2 mM) (including its biological concentration range, i.e. 0.75-0.90 mM), increasing the Mg2+ concentration increased the fraction of free phloroglucinol (not linked with HSA) and thus its biological effect. PMID- 14565561 TI - A novel chemiluminescence method for determination of terbutaline sulfate based on potassium ferricyanide oxidation sensitized by rhodamine 6G. AB - This work reports a novel flow injection-chemiluminescence (FI-CL) system for determination of terbutaline sulfate, a drug for treatment of asthma and chronic obstructive pulmonary disease (COPD). It is based on the reaction of potassium ferricyanide with terbutaline sulfate in sodium hydroxide medium sensitized by the fluorescent dye rhodamine 6G. With the peak height as a quantitative parameter applying optimum working conditions, terbutaline sulfate is determined over the range of 0.01-1.2 microg ml(-1) with a detection limit of 6.7 x 10(-3) microg ml(-1). The relative standard deviation (R.S.D.) is 3.7% for 0.1 microg ml(-1) terbutaline sulfate (n = 11). The proposed method is sensitive, simple, rapid, and was applied to determination of terbutaline sulfate in pharmaceutical preparations. The possible chemiluminescence (CL) reaction mechanism was also discussed briefly. PMID- 14565562 TI - A simple rapid method of sample preparation for LC analysis of retinoyl beta glucuronide and retinoic acid in water-based creams. AB - A simple method of sample preparation for LC analysis of retinoic acid (RA) and retinoyl beta-glucuronide (RAG) in creams has been developed. Water-based cream of all trans-RAG, devoid of side effects but efficacious in the treatment of acne vulgaris, was found to be hydrolyzed to RA in a temperature dependant manner. The potential benefits of water-based RAG cream stored at room temperature for the treatment of acne and wrinkle is discussed. PMID- 14565563 TI - Troubleshooting problems with in vitro screening of drugs for QT interval prolongation using HERG K+ channels expressed in mammalian cell lines and Xenopus oocytes. AB - The majority of drugs associated with QT interval prolongation share an ability to inhibit ionic currents passed by HERG potassium channels. One method of screening new chemical entities (NCEs) for QT prolonging potential is therefore to use heterologous systems expressing HERG channels. Such systems are also of value in the understanding of the function, kinetics, sorting, pharmacological sensitivities, and important molecular determinants of the HERG potassium channel. The methods for incorporating the HERG potassium channel into cells and measuring the consequent current are a mixture of techniques that are standard (for heterologous expression of most ion channels) and individualised to HERG. This review presents a selection of the most commonly used methods for examining heterologous HERG currents, as well as introducing some of the technical problems that may be encountered and their solutions. In mammalian cell lines, problems such as fragile membranes, high leak currents, inability to form a gigaseal, diminished HERG current, endogenous transient outward current, altered kinetics, and even occasional run down can interfere with measurements. In Xenopus oocytes, endogenous chloride currents, insufficient superfusate flow, diminished HERG current and HERG current 'run up' may create difficulties. PMID- 14565564 TI - An improved method for measuring vascular permeability in rat and mouse skin. AB - INTRODUCTION: We have improved a rodent vascular permeability measurement method employing fluorescent dye-labeled bovine serum albumin. METHODS: The incubation duration for direct fluorescent detection of skin injected with an inflammatory agent was decided based on regression curve parameters with the correlation coefficient obtained from the least squares method. RESULTS: A suitable incubation time was determined to be 2-6 h. The recovery of FITC-BSA from the skin sample was very good, and the correlation coefficient of the linear regression curve was .99. The linear relation between the previous dye extraction method using brilliant blue 6B and the new and improved fluorescence method was very high. In mice, histamine-induced serum exudation in the back skin increased from 0.31 to 1.25 microg/site in a dose-dependent manner and reached a plateau at 1.25-2.5 microg/site. The serum exudation caused by histamine increased to 10 microg/site and almost reached a plateau at 10-40 microg/site in rats. The time required for the measurement of fluorescence intensity was very short because a microplate reader was used as the measurement apparatus. CONCLUSION: The improved method is easy to use and sensitive and does not necessitate extraction of dye from the skin. PMID- 14565565 TI - Quantification of endothelins, their receptors, and endothelin-converting enzyme mRNAs in rat genitourinary tract using real-time RT-PCR. AB - INTRODUCTION: Quantification of mRNA expression is essential for the assessment of endothelin (ET) receptor-mediated mechanisms. Recently, a novel technique for the determination of mRNA expression, termed real-time reverse transcription polymerase chain reaction (RT-PCR), has been developed. We therefore applied real time PCR using SYBR Green I to quantify ET-1, ET-3, ET-converting enzyme-1 (ECE 1), and ETA and ETB receptor subtype mRNA expression in the rat genitourinary tract. METHODS: The cDNA was synthesized by RT of RNA extracted from the rat bladder, ventral prostate, dorsolateral prostate, and vas deferens. All steps subsequent to the RT reaction were carried out by the thermal cycler/detector and computer-assisted programs processed a quantitative result. RESULTS: Designing optimal primer sequences that minimized primer-dimer formation and adjusting annealing temperatures that prevented nonspecific product amplification have made it possible to identify a single peak in the melt curve and to obtain an appropriate standard curve for each gene transcript. In our experiments, input cDNA levels as low as 100 copies of the product could be detected. DISCUSSION: We demonstrated that significant quantitative variations existed in the expression levels of ET-1, ET-3, ECE-1, and ETA and ETB receptor subtype mRNAs within a tissue and between different regions of the genitourinary tract and that the predominant expression of ETs and their receptor mRNAs in all tissues studied were ET-1 and the ETA receptor subtype, respectively. PMID- 14565566 TI - Site-specific absorption of M100240 and MDL 100,173 in rats evaluated using Sweetana-Grass diffusion chamber technology. AB - INTRODUCTION: M100240--an acetate thioester of MDL 100,173--a dual angiotensin converting enzyme (ACE)/neutral endopeptidase (NEP) inhibitor is in Phase II development for cardiovascular conditions. The absorption characteristics of M100240 and MDL 100,173 were evaluated in specific regions of the rat gastrointestinal (GI) tract. METHODS: Solutions of M100240 and MDL 100,173 were prepared using mucosal Kreb's solution. Four male Sprague-Dawley rats, fasted before tissue procurement, were sacrificed using 95% CO2 and 5% O2. The entire small and large intestine were removed, rinsed in serosal Kreb's solution, and segments mounted onto the Sweetana-Grass diffusion chamber. Test compounds were added to the donor compartment and drug-free oxygenated serosal Kreb's solution to the receiving compartment. The temperature of the chambers was maintained at 37 degrees C and supplied with 95% O2/5% CO2. Samples were removed (0.5 ml) at 0, 30, 60, 90, 120, 180, and 240 min from the serosal side. The volume was maintained with drug-free warm serosal Kreb's solution. Samples were diluted with rabbit plasma and analyzed by liquid chromatography/mass spectrometry. Apparent permeation values [Papp (cm/min)] of M100240 and MDL 100,173 through the duodenum, jejunum, ileum, and colon were determined. RESULTS: The mean Papp of M100240 and MDL 100,173 was highest in the duodenum: 2.29 x 10(-4) +/- 2.40 x 10( 4) and 1.66 x 10(-4) cm/min +/- 8.33 x 10(-5), respectively. The mean Papp was lowest in the colon: 3.61 x 10(-6) +/- 3.44 x 10(-6) and 1.62 x 10(-5) +/- 3.21 x 10(-6) cm/min for M100240 and MDL 100,173, respectively. Absorption of MDL 100,173, however, was evident throughout the rat GI tract. DISCUSSION: M100240 and MDL 100,173 are predominantly absorbed from the duodenum in the rat GI tract. MDL 100,173 is also absorbed from the jejunum, ileum, and colon. These results, consistent with data obtained in humans, demonstrate the potential predictive value of the Sweetana-Grass model for site of absorption assessments. PMID- 14565567 TI - Organotypic liver culture in a fluid-air interface using slices of neonatal rat and adult human tissue--a model of fibrosis in vitro. AB - INTRODUCTION: Fibrosis is the common end stage of most liver disease but there is no effective treatment currently available. We hypothesised that if viability of liver tissue slice culture could be improved, it should be possible to develop a model of liver fibrosis in vitro that could advance the development of antifibrotic therapy while at the same time reducing the need to use in vivo models. We have adapted a slice culture technique developed originally for organotypic culture of neural tissue to the liver. METHODS: slices of neonatal rat or adult human liver, 100-400-microm thick, were cut and cultured on nitrocellulose inserts at the air/fluid interface for up to 28 days. RESULTS: Hepatocytes expressed albumin by immunocytochemistry for up to 10 days and were viable for up to 21 days during which time new structures appeared, including cytokeratin 19 positive bile ductular structures and bands of smooth muscle actin positive stellate cells associated with new reticulin positive matrix. Smooth muscle actin expression by stellate cells could be pharmacologically inhibited by SDZ-RAD (everolimus). DISCUSSION: In conclusion, we have successfully developed a novel model of liver culture, which may prove useful in both studies of the mechanisms of liver fibrosis and in developing therapeutic strategies. PMID- 14565568 TI - An automated coronary artery occlusion device for stimulating collateral development in vivo. AB - INTRODUCTION: Repetitive, brief coronary artery occlusions produce collateral development in experimental animals. This model causes coronary collateralization in a highly reproducible fashion, but the process is very labor intensive. We report the design and use of a fully automated hydraulic coronary occlusion device capable of producing repetitive coronary occlusions and enhancement of coronary collateral development in dogs. METHODS: The device consists of analog electronics that allow adjustment of occlusion number, frequency, pressure and duration, and mechanical components responsible for the coronary occlusion. The motor and piston of the device are coupled to a chronically implanted hydraulic vascular occluder placed around the left anterior descending coronary artery (LAD) of dogs instrumented for measurement of systemic and coronary hemodynamics. One group of dogs (n = 6) underwent brief (2 min) LAD occlusions once per hour, eight times per day, 5 days/week for 3 weeks to stimulate collateral development (measured using radioactive microspheres). Another group of dogs (n = 6) that did not receive repetitive occlusions served as controls. RESULTS: The device reproducibly produced repetitive LAD occlusions for the duration, frequency, and time interval initially programmed. A time-dependent increase in transmural collateral blood flow was observed in dogs undergoing repetitive occlusions using the device. Collateral blood flow was unchanged in dogs that did not undergo occlusions. DISCUSSION: The automated occluder device reliably produces repetitive coronary occlusions and may facilitate further study of coronary collateral development in response to chronic myocardial ischemia. PMID- 14565569 TI - Coincubation of tissue slices, a new way to study metabolic cooperation between organs: hepatorenal cooperation in the biotransformation of CGP 47 969 A. AB - INTRODUCTION: There is limited information on in vitro/ex vivo tools to be used for studying interorgan metabolic cooperation. We report here the use of the tissue slice technique for this purpose. METHODS: Rat liver and kidney slices were used to study metabolic cooperation for the metabolism of CGP 47 969, a potential anti-inflammatory compound which in vivo is extensively conjugated with glutathione and subsequently degraded via the mercapturic acid pathway. RESULTS: Upon incubation with liver slices, CGP 47 969 was extensively conjugated with GSH whilst degradation of the GSH conjugate was moderate. Upon incubation with kidney slices, conjugation of CGP 47 969 with GSH was moderate but degradation of the GSH conjugate was complete. Upon coincubation of CGP 47 969 with liver and kidney slices, both conjugation with GSH and its subsequent degradation were almost complete. Thus, coincubation of liver and kidney slices permitted the efficient in vitro reproduction of the complete biotransformation of CGP 47 969 via its GSH conjugate to the ultimate mercapturic acid metabolite in a one step procedure. DISCUSSION: This novel slice coincubation culture could serve as an in vitro model for interorgan cooperation in multistep metabolic processing. PMID- 14565570 TI - Limitations of whole-body autoradiography without receptor microautoradiography. PMID- 14565572 TI - Effects of copper on wild and tolerant strains of the lichen photobiont Trebouxia erici (Chlorophyta) and possible tolerance mechanisms. AB - The influence of copper was assessed on wild and tolerant strains of the lichen photobiont Trebouxia erici and shown to have multiple toxic effects. Addition of 4 mM copper chloride into Trebouxia media reduced growth rates of the wild type to less than 1% of control levels. It also injured cell membranes, inhibited dehydrogenase activity, altered pigment composition, and reduced activity of photosystem II. Membrane damage was assessed through measuring electrical conductivity and potassium content, dehydrogenase activity by degree of conversion of 2,3,5-triphenyl tetrazolium chloride to red triphenylformazan, and functioning of PS II by chlorophyll a fluorescence. In respect to most parameters the tolerant strain was usually less affected by copper than the wild strain and, in some cases, not at all. We demonstrated by inductively coupled plasma atomic emission spectrometry and energy-dispersive X-ray microanalysis that enriched copper in the medium resulted in increased uptake by both wild and tolerant photobiont cells, but extracellular concentrations were significantly higher in the tolerant strain. This suggested that, at least in vitro, extracellular deposition was a mechanism of copper tolerance. PMID- 14565571 TI - Perfluorooctane sulfonate concentrations in surface water in Japan. AB - Perfluorooctane sulfonate (PFOS) is a class of specialty chemicals used in a variety of applications, and has been found to be globally distributed in many living organisms including humans. Several analytical methods have been developed for determination of PFOS in environmental samples and biological matrices. However, these methods employ liquid chromatography/tandem mass spectrometry (LC/MS/MS), an instrumentation which has limited accessibility because it is expensive to use and maintain. In the present study we present the development of a robust analytical method using liquid chromatography/mass spectrometry (LC/MS) in combination with solid phase extraction. The high yield and concentration of the present method enabled us to quantify PFOS as low as 0.1 ng/L. This method was applied to the determination of PFOS in 142 surface water samples collected from various geographic locations around Japan. The geometric mean (geometric standard deviation) (ng/L) for river samples (n = 126) was 2.37 (4.13), with a median of 1.68 and a range of 0.3-157 ng/L, and for coastal sea water samples (n = 16) was 1.52 (4.14), with a median of 1.21 and a range of 0.2-25.2 ng/L. However, the concentrations in most of the samples were much lower than the values reported in the US, except for those from the Jinzu (135.0 ng/L) and Tama (157 ng/L) Rivers. Because surface waters in the Ara (13.0-38.5 ng/L), Tama (0.7 157.0 ng/L), and Yodo (0.9-27.3 ng/L) Rivers, sources of drinking water for more than eight million people, were moderately contaminated with PFOS, more work is needed to assess exposure to PFOS. PMID- 14565573 TI - Peroxidase and superoxide dismutase activities in fig leaves in response to ambient air pollution in a subtropical city. AB - Urban air pollution is a serious problem in both developing and developed countries, and antioxidant enzyme activities in plants have been suggested as a useful bioindicator of air pollution. In this study, the seasonal and spatial variability of peroxidase and superoxide dismutase activities were measured in leaves of Ficus microcarpa at eight sampling sites in the Taipei metropolitan area and one background site in rural area at each month for a year. The spatial pattern of peroxidase activity in figs collected from the Taipei metropolitan area was similar to the spatial pattern of O3 concentration in the Taipei metropolitan area. The peroxidase activities of Ficus microcarpa were significantly higher at sampling sites from the outer zone of the metropolitan area than those from the inner zone of the metropolitan area in spring and summer. On the other hand, the spatial pattern of superoxide dismutase activity in fig leaves did not show significant differences between the inner and outer zones of the Taipei metropolitan area. In addition, peroxidase activities, but not superoxide dismutase activities, of Ficus microcarpa were significantly higher in sites with high traffic density than those in low traffic density sites. Even though peroxidase activities in Ficus microcarpa tended to be higher in high traffic density sites or some sites with high ozone concentration, site specific changes of peroxidase activity in Ficus microcarpa due to O3 pollution were not clearly observed in this study. Based on these results, neither peroxidase nor superoxide dismutase in Ficus microcarpa is a sensitive bioindicator for O3 pollution, although peroxidase shows some potential to be used as a general bioindicator of air quality. PMID- 14565574 TI - Determination of lipid peroxides in invertebrates tissues using the Fe(III) xylenol orange complex formation. AB - Reactive oxygen species (ROS) are subproducts of the oxidative metabolism known to initiate chain reactions with polyunsaturated fatty acids that generate lipid peroxides (LPO). The objective of this work was to adapt the ferrous oxidation/xylenol orange (FOX) assay to measure LPO in invertebrate tissues i.e.: from polychaeta (Laeonereis acuta) and crab (Chasmagnathus granulata) species. Whole polychaetes were homogenized in methanol 100%, being determined the optimal sample volume and the time required for color development. It was tested five sample volumes (8-30 microl), following color development up to 215 min. Absorbance stabilization was observed after 90 min, being linearly related with sample volume. A similar procedure was adopted for crab tissues (anterior gills, posterior gills, and hepatopancreas). Differences between species and between organs of the same species were observed when analyzed nonspecific absorbance increments after adding the standard cumene hydroperoxide (CHP). In polychaeta and crab anterior gills tissue, absorbance increments were lower (21-25%) than samples without tissue extracts (blanks) that received CHP. In crab posterior gills and hepatopancreas, the nonspecific increment was almost negligible. Correction formulae are given to account for these differences and simplified protocols for each tissue and species are also included. Great differences in the lipid peroxides content was detected between worms (127.05 +/- 19.32 nmoles CHP/g of wet tissue) respect to anterior gills, posterior gills, and hepatopancreas from the crab species (52.65 +/- 3.59, 30.54 +/- 4.73, and 48.51 +/- 8.78 nmoles CHP/g of wet tissue, respectively). PMID- 14565575 TI - Mercury toxicity in the aquatic oligochaete Sparganophilus pearsei: I. Variation in resistance among populations. AB - Mercury contamination has become a problem in many San Francisco Bay Area watersheds due to its elevated presence in sediments and aquatic organisms. The present study used laboratory lethal toxicity (LC50) tests to examine the mercury tolerance of aquatic oligochaete worms, Sparganophilus pearsei, from contaminated and uncontaminated areas. The oligochaetes were collected in the following fresh water reservoirs: Sandy Wool (reference area), San Pablo, Lake Anza, Lake Herman, and Guadalupe. These last four reservoirs were contaminated with levels of mercury that ranged from 1.5 to 2 mg/kg (wet weight). Mercury concentrations in sediment and tissue from Sandy Wool were below detection limits and worms from this site were the least tolerant of mercury in laboratory exposures (LC50 = 0.22 mg/L). Worms from the other, more contaminated, reservoirs contained elevated tissue mercury concentrations and were more tolerant in laboratory tests (LC50 = 1.48-2.19 mg/L). The present study demonstrates that different populations of the aquatic oligochaete S. pearsei have developed different tolerances to mercury depending on their previous history of exposure to mercury contamination. PMID- 14565576 TI - Echinoderms as bioindicators, bioassays, and impact assessment tools of sediment associated metals and PCBs in the North Sea. AB - The study assessed the occurrence, possible toxicity, and impact of sediment associated metals and PCBs in the coastal zone of the southern North Sea using echinoderms as representatives of the macrobenthos. Metals and PCBs were analyzed in the sediments and in the body compartments of the starfish Asterias rubens from 11 stations. The general toxicity of sediment-associated contaminants was assessed by bioassays using embryonic and larval developments of both A. rubens and the sea urchin Psammechinus miliaris. The impact of contamination was assessed by measuring cellular immune responses of A. rubens collected in the same stations. Contamination of the starfish by metals and PCBs closely reflected that of the sediments. However, bioaccumulation was element-specific for metals and depended on the chlorination pattern for PCBs. The sediment-associated contaminants appeared to be toxic in both the A. rubens and P. miliaris developmental assays. Moreover, metals were shown to affect the immune responses of starfishes living in contaminated stations. The most significant effects on biological responses were recorded in the plumes of the Scheldt/Rhine/North Sea Canal and the Elbe/Weser Rivers. PMID- 14565577 TI - Reduction in the sex ratio of the progeny of a parasitoid wasp (Trichogramma brassicae) surviving the insecticide chlorpyrifos. AB - Due to the wide use of insecticides and their contribution to environmental pollution, the determination of their sublethal effects on the reproduction of parasitoids regulating pest populations is a necessity. In this work, sublethal effects of a lethal dose 20% of chlorpyrifos on the reproduction and the sex ratio of Trichogramma brassicae were assessed. The organophosphorus insecticide chlorpyrifos is one of the most used insecticides for agricultural and domestic purposes in a worldwide market. A decrease in the number of parasitized hosts was observed for females surviving the insecticide. Furthermore, while untreated females presented a very biased sex ratio in favor of daughters, the offspring of treated females surviving the insecticide presented an almost balanced sex ratio due to a decrease in the number of daughters. This decrease in sex ratio was interpreted to be resulting from a decrease in fertilized eggs due to the perturbation of nerve transmissions induced by the insecticide. These results highlight the need for considering not only the direct mortality induced by insecticides but also sublethal effects liable to modify important fitness traits involved in the dynamic of beneficial insect populations. PMID- 14565578 TI - Influences of a laboratory diet and natural seston on the bioavailability of carbaryl, chlorpyrifos, and malathion to black fly larvae (Diptera: Simuliidae) in an acute toxicity test. AB - To determine the effects organic food particles can have on insecticide bioavailability, two food types, a laboratory diet and dried, natural seston, were evaluated using black fly larvae, Simulium virtatum Zetterstedt cytospecies IS-7, in a 24-h orbital shaker toxicity test. The bioavailability of all three insecticides was significantly altered when diet concentrations in the flasks were > or = 150 mg/L. Chlorpyrifos availability decreased, whereas carbaryl and malathion availability increased. Dried, natural seston had little effect on the bioavailability of these insecticides except in carbaryl-dosed larvae where mortality was significantly increased when seston concentrations were 150 mg/L in the flasks. Differences in insecticide bioavailability between the two food types in these experiments might have been related to the particle size and organic carbon content of the two materials. Laboratory diet particles were significantly smaller (11.7 +/- 0.5 microm) and had a significantly greater organic carbon content (26.4%) than the seston particles (30.9 +/- 3.3 microm; 1.1%). These results suggest that the concentration, as well as the physical and chemical component of the food source, are important factors in determining the effects of food on insecticide bioavailability in aquatic systems. PMID- 14565580 TI - Effects of the insecticide endosulfan and presence of congeneric tadpoles on Australian treefrog (Litoria freycineti) tadpoles. AB - The organochlorine insecticide endosulfan is extensively used in Australia and can often contaminate aquatic environments. However, its effects on Australian frog species are unknown. We exposed tadpoles of the Australian treefrog Litoria frevcineti to endosulfan for 96 hours. Tadpoles exposed to 0.03 or 1.3 microg/L endosulfan grew more slowly than control tadpoles. Furthermore, feeding was inhibited in L. freycineti when the animals were observed halfway through their exposure to a concentration of 1.3 microg/L endosulfan for 96 h. This concentration (which periodically occurs in natural water bodies and rivers) also caused 17% tadpole mortality, and the survivors were more vulnerable to invertebrate (odonate) predation when tested 15 days after transfer to clean water. The presence of a sympatric species of tadpole, Litoria peronii, did not affect survivorship of L. freycineti, but reduced growth rates. Thus, short-term exposure to endosulfan in natural water bodies may influence tadpole viability either immediately or over an extended period. PMID- 14565579 TI - Impact of naled on honey bee Apis mellifera L. survival and productivity: aerial ULV application using a flat-fan nozzle system. AB - A study was conducted to evaluate the impact of naled on honey bees as a result of their exposure to aerial ULV applications of this insecticide during three routine mosquito spray missions by Manatee County Mosquito Control District in Florida during the summer of 1999. Naled deposits were collected on filter paper and subsequently analyzed by gas chromatography. Mortality of adult honey bees Apis mellifera L. was estimated based on numbers from dead bee collectors placed in front of the entrance of the beehives. We found that honey bees clustering outside of the beehives were subject to naled exposure. Bee mortality increased when higher naled residues were found around the hives. The highest average naled deposit was 6,227 +/- 696 microg/m2 at the site 1 forest area following the mosquito spray mission on July 15, 1999. The range of naled deposition for this application was 2,818-7,101 microg/m2. The range of dead bees per hive was 0-39 prior to spraying and 9-200 within 24 h following this spray mission. The average yield of honey per hive was significantly lower (p < 0.05) for naled-exposed hives compared with unexposed hives. Because reduction of honey yield also may be affected by other factors, such as location of the hives relative to a food source and vigor of the queen bee, the final assessment of honey yield was complicated. PMID- 14565581 TI - Comparing polybrominated diphenyl ether and polychlorinated biphenyl bioaccumulation in a food web in Grand Traverse Bay, Lake Michigan. AB - Levels of polybrominated diphenyl ethers (BDEs) in Great Lakes salmonids and ambient air have been recently reported, but few studies worldwide have examined the accumulation of BDEs within aquatic food webs. Here we report some of the first measurements of six BDE congeners that are common components of the pentaBDE commercial mixture within an entire Lake Michigan food web. BDEs were detected in all samples and the dominant BDE congener was 2,2',4,4' tetrabromodiphenyl ether (BDE 47). BDE 47 levels were consistently greater than those of the 2,2',4,4',5-pentabromodiphenyl ether (BDE 99), despite similar levels of these two compounds in commercial mixtures, suggesting differences in the bioavailability of the BDE congeners or differences in their ability to be metabolized. Additionally, congener composition was significantly different among deepwater sculpin, bloater chub, and lake trout, indicating differences in exposure or differences in biotransformation capacities. Total BDE concentrations in this food web were positively correlated (r = 0.94) with levels of PCBs previously measured in these samples (Stapleton et al. 2001a). Levels of BDE 47 and PCB 153, compounds with similar physicochemical properties, were compared to examine the relative exposure and bioaccumulation of these two classes of chemicals that have different environmental loading histories. Food web magnification factors calculated for these two congeners were 3.2 and 4.0 for BDE 47 and PCB 153, respectively, indicating a comparable potential for biomagnification in food webs. PMID- 14565582 TI - Mercury biomagnification in a tropical black water, Rio Negro, Brazil. AB - The population living along the riverbanks of the Amazon basin depends heavily on fish for nutritional support. Mono-methyl-mercury (MMHg) concentrates in fish, which can contaminate humans, the risk depending not only on fish MMHg concentration but also on the amount of fish consumed. We sampled nine locations of the Rio Negro basin, differing in water pH, Hg concentrations, and dissolved organic carbon (DOC), and determined total Hg from 951 fish samples of species representative of the food web: herbivorous, detritivorous, omnivorous, and piscivorous. Mercury concentrations varied widely in all species but showed a trend that depended on fish feeding strategies. The highest mean concentration was found in the piscivorous species (688.90 ng/g(-1)), followed by omnivorous (190.30 ng/g(-1)), detritivorous (136.04 ng/g(-1)), and herbivorous (70.39 ng/g( 1)). Fish Hg concentrations exceeding current safe limits (500 ng/g(-1)) for human consumption were found mainly in the piscivorous species (60%). Significant positive correlation between fish weight and Hg concentration was seen for the piscivorous Serrasalmus spp. (n = 326; r = 0.3977; p < 0.0001), Cichla spp. (n = 125; r = 0.4600; p < 0.0001), and Pimelodus spp. (n = 12; r = 0.8299; p = 0.0008), known locally as Piranha, Tucunare, and Mandi, respectively. However, a negative correlation was seen for non-piscivorous Potamorhina latior (n = 30; r 0.3763; p = 0.0404) and Leporinus spp. (n = 44; r = -3987; p = 0.0073), known as Branquinha (detritivorous) and Aracu (omnivorous). Fish-Hg concentrations in the acidic waters (pH range, 4.09-6.31) of the Rio Negro habitat, with its wide gradient of Hg concentrations (3.4-11.9 microg/L(-1)) and DOC (1.85-15.3 mg/L( 1))--but no history of gold mining activity-are comparable to other Amazonian rivers. Opportunity fish catches in the Rio Negro habitat show high muscle-Hg derived from natural sources, but no systematic association with site-dependent geochemistry. PMID- 14565583 TI - Exposure to water from the lower Rhine induces a stress response in the rainbow trout Oncorhynchus mykiss. AB - The water quality of the river Rhine has improved in recent years and populations of salmonids are increasing. Nevertheless at present, the water from the lower Rhine still contains a complex mixture of low levels of many pollutants and it is not known whether exposure to such water is stressful to salmonid fish. For 31 days we continuously exposed the trout Oncorhynchus mykiss to water from the lower Rhine in the Netherlands and measured a variety of physiological, biochemical, and histological parameters, including the stress parameters cortisol and glucose. Exposure to Rhine water significantly increased cortisol and glucose after 3 h. At 21 and 31 days, cortisol was lower in exposed fish, indicating inhibition or exhaustion of the hypothalamic-pituitary-interrenal (HPI) axis. Electron microscopical analysis of the skin and gill epithelia revealed stressor-related effects that reflected disruption of the skin epithelium, the interface between the fish and the environment. This had little influence on hydromineral balance, as neither gill Na+/K+-ATPase activity nor plasma Na+ and Cl- were altered, although intestine- and kidney-specific Na+/K+ ATPase activities were affected. Analysis of heavy metal concentrations in the liver, kidney, and intestine indicated no bioaccumulation. Immunostimulation was reflected by increased respiratory burst activity of the head kidney leukocytes. From 7 days onwards, the body weight of the Rhine water fish was significantly lower than that of control fish. Overall, the data show that acute exposure to present day water from the lower Rhine induced a stress response in the fish that, during chronic exposure, was followed by impairment of the HPI axis, reduced growth, and prolonged immunostimulation. PMID- 14565584 TI - Toxicity of chlorpyrifos adsorbed on humic colloids to larval walleye (Stizostedion vitreum). AB - After application, organophosphorus insecticides (OPs) are often strongly adsorbed to soil constituents. Because of their relatively low water solubility, OPs may be transferred from field to stream adsorbed on suspended solids. However, we are not aware of research done to evaluate the bioavailability (i.e., toxicity) of OPs transported on suspended solids to fish. We conducted 48-h static toxicity tests to determine the toxicity of chlorpyrifos in aqueous solution and adsorbed on calcium-saturated humic acid (HA) to three larval stages of walleye (Stizostedion vitreum). Three concentrations of chlorpyrifos adsorbed on HA, a HA control, and a chlorpyrifos-only treatment were tested. Fish that survived the 48-h static toxicity tests were analyzed to determine total cholinesterase (ChE) activity. In general, survival of all larval stages of walleye exposed to chlorpyrifos-HA complexes was less than that of walleye exposed to HA controls and the chlorpyrifos-only treatment, which were not toxic to walleye. Cholinesterase inhibition of larval walleye exposed to chlorpyrifos HA complexes was similar to the ChE inhibition observed in larval walleye exposed to chlorpyrifos in the aqueous phase. These laboratory experiments indicate potential toxicity of chlorpyrifos-soil complexes to larval fish. PMID- 14565585 TI - Assessment of human health risks for arsenic bioaccumulation in tilapia (Oreochromis mossambicus) and large-scale mullet (Liza macrolepis) from blackfoot disease area in Taiwan. AB - This paper carries out probabilistic risk analysis methods to quantify arsenic (As) bioaccumulation in cultured fish of tilapia (Orechromis mossambicus) and large-scale mullet (Liza macrolepis) at blackfoot disease (BFD) area in Taiwan and to assess the range of exposures for the people who eat the contaminated fish. The models implemented include a probabilistic bioaccumulation model to account for As accumulation in fish and a human health exposure and risk model that accounts for hazard quotient and lifetime risk for humans consuming contaminated fish. Results demonstrate that the ninety-fifth percentile of hazard quotient for inorganic As ranged from 0.77-2.35 for Taipei city residents with fish consumption rates of 10-70 g/d, whereas it ranged 1.86-6.09 for subsistence fishers in the BFD area with 48-143 g/d, consumption rates. The highest ninety fifth percentile of potential health risk for inorganic As ranged from 1.92 x 10( 4)-5.25 x 10(-4) for Taipei city residents eating tilapia harvested from Hsuehchia fish farms, with consumption rates of 10-70 g/d, whereas for subsistence fishers it was 7.36 x 10(-4)-1.12 x 10(-3) with 48-143 g/d consumption rates. These findings indicate that As exposure poses risks to residents and subsistence fishers, yet these results occur under highly conservative conditions. We calculate the maximum allowable inorganic As residues associated to a standard unit risk, resulting in the maximum target residues, are 0.0019-0.0175 and 0.0023-0.0053 microg/g dry weight for tilapia and large-scale mullet, respectively, with consumption rates of 70-10 g/d, or 0.0009-0.0029 and 0.0011-0.0013 microg/g dry weight for consumption rates of 169-48 g/d. PMID- 14565586 TI - Maternal transfer of contaminants to eggs in common grackles (Quiscalus quiscala) nesting on coal fly ash basins. AB - Coal combustion is a major source of trace elements to the environment. Aquatic disposal of wastes from this process can result in reproductive failure in many wildlife species, but little is known regarding impacts on avian fauna. Individual eggs were collected from common grackles (Quiscalus quiscala) nesting in association with coal fly ash settling basins and a reference site to determine if females from the contaminated site transfered trace elements to their eggs. Whole clutches were also collected from both sites to examine inter- and intra-clutch variability of maternally transferred contaminants. Selenium was the only trace element found in significantly higher concentrations in ash basin eggs (x = 5.88 +/- 0.44 microg/g DW) than in reference eggs (x = 2.69 +/- 0.13 microg/g DW). Selenium concentrations in eggs from the ash basins were above background levels, but did not exceed higher proposed Se toxicity thresholds. Inter- and intra-clutch variation was higher for ash basin clutches than reference clutches. The relationship between selenium concentrations and laying order (estimated by egg mass) was not statistically significant, but increased Se concentration in the second egg of most ash basin clutches followed by declining concentrations in subsequent eggs suggested that further examination of this pattern might be warranted. PMID- 14565587 TI - Placental cadmium and lipid peroxidation in smoking women related to newborn anthropometric measurements. AB - Smoking during pregnancy is known to be harmful for fetal growth, and cigarette smoke is the major source of cadmium exposure in the general population. Some authors suggest that cadmium accumulated in placenta may be the main cause of the decreased fetal growth shown by infants born to smoking mothers. The aim of our study was to assess any relationship between smoking during pregnancy, placental cadmium levels, placental lipid peroxidation, and anthropometric measurements of newly born babies. The mean values of placental cadmium were significantly higher in smoking mothers, who also produced offspring with reduced neonatal weight, length, head and abdominal circumferences. Lipid peroxidation correlated negatively with birth weight and abdominal circumference. Our results suggest that the cadmium accumulated in placenta is not the reason for reduced fetal growth in smoking mothers and that placental peroxidation is not enhanced by smoking, although the outcome of pregnancy seems to be negatively influenced by lipid peroxidation. PMID- 14565588 TI - Kinetic transport of pesticide from contaminated fabric through a model skin. AB - Most protective clothing research for workers exposed to pesticides has focused on the primary dermal exposure that results from direct contact with the pesticide and on the penetration, versus repellency, of protective materials. The role of absorption and retention in the function of traditional clothing materials to limit pesticide exposure has received less attention. This project investigates the transfer of pesticides from contaminated work clothing in the dry state to human skin with the goal of furthering our understanding of the role of absorption and retention in the function of traditional clothing materials. Our objective was to study the use of synthetic membrane to evaluate the kinetic transport of pesticide from contaminated clothing through human skin. Transport of pesticide through the test system with and without the presence of fabric was characterized by a three-parameter single exponential rise to maximum equation. The synthetic membrane system was an appropriate model for human skin in this situation. Starching applied to the fabric gave additional absorption and retention, reducing pesticide transport from the fabric to the skin, and heavier weight fabric, such as denim, provided protection through absorption and retention of the pesticide. PMID- 14565589 TI - Inflammatory mediators as essential elements in bone remodeling. AB - Inflammatory disorders such as rheumatoid arthritis (RA), may have profound effects on skeletal homeostasis. In contrast to physiologic remodeling in which mechanical influences and/or systemic endocrine hormones initiate the remodeling process, in disorders such as RA the recruitment of macrophage lineage cells to sites of inflammation and the action of local osteoclastogenic cytokines associated with the inflammatory process initiate the remodeling process. In both physiologic and pathologic remodeling, osteoclasts appear to be the principal cell type responsible for the bone resorption. In addition, many of the same cytokines and mediators are involved in physiologic and pathologic bone remodeling. These observations have important implications with respect to the development of therapeutic strategies to prevent bone loss in inflammatory conditions. PMID- 14565590 TI - Fatigue microdamage as an essential element of bone mechanics and biology. AB - The fossil record shows that bone remodeling has existed since the earliest large vertebrates became weight-bearing on land, but the functions of remodeling have long been debated. The principal protagonists in this debate have been those favoring a mechanical function and those asserting that remodeling serves to move calcium in and out of the skeleton. In recent years the arguments of the former school have included not only the adaptation of internal structure to specific kinds of stresses, but the need to remove fatigue damage. It has become clear that (1) physiologic strains continually produce fatigue damage in bone; (2) this damage weakens bone and is associated with both osteocyte apoptosis and the activation of remodeling; and (3) remodeling is the only means by which this damage can be removed. The significance of these observations is increased by the fact that fatigue failure is more likely in larger structures. This "volume effect," along with the advantages of enhanced mobility and metabolic efficiency, may have selected for bone remodeling as a means of controlling fatigue damage as it occurs, allowing larger vertebrates to maintain a relatively light skeleton over an extended lifetime. In this view, bone remodeling is not primarily a mechanism for calcium transport, but is intimately related to other inflammatory repair responses. PMID- 14565591 TI - A 3-year longitudinal study of the effect of physical activity on the accrual of bone mineral density in healthy adolescent males. AB - It has previously been suggested that physical activity predominantly influences the accumulation of bone density before puberty. The purpose of the present study was to examine the effect of physical activity on the accumulation of bone mass in male athletes between 16 and 19 years of age. The cohort studied consisted of 12 badminton players (aged 16.1 +/- 0.5), 20 ice hockey players (aged 16.1 +/- 0.5), and 24 age-matched controls (aged 16.1 +/- 0.6). The bone mineral density (BMD, g/cm2) of the total body, spine, dominant and nondominant humerus, head and femoral neck was measured twice with a 3-year interval by dual energy X-ray absorptiometry (DXA). In addition, at the femoral neck, volumetric bone mineral density (vBMD, mg/cm3) was estimated. At baseline, the athletes as a whole group had significantly higher BMD at the total body (P = 0.03), dominant (P = 0.006) and nondominant humerus (P = 0.009) and femoral neck (P = 0.007) compared to the controls. At the 3-year followup, the athletes had significantly higher BMD at all sites (total body; P = 0.003, spine; P = 0.02, dominant humerus; P = 0.001, nondominant humerus; P = < 0.001, femoral neck; P = 0.001) except for the head (P = 0.91) compared with controls. The athletes also had higher vBMD at the femoral neck compared with the controls (P = 0.01). Furthermore, to be an athlete was found to be independently associated with a higher increase in nondominant humerus BMD (beta = 0.24; P < 0.05) and femoral neck BMD (beta = 0.30; P < 0.05) compared with the controls, during the study period. In summary, these results suggests that it is possible to achieve continuous gains in bone mass in sites exposed to osteogenic stimulation after puberty in males by engaging in weight bearing physical activity. PMID- 14565592 TI - Pattern of periprosthetic bone remodeling around stable uncemented tapered hip stems: a prospective 84-month follow-up study and a median 156-month cross sectional study with DXA. AB - Bone resorption in the proximal femur is commonly seen after total hip arthroplasty (THA). With dual energy X-ray absorptiometry (DXA), the amount of bone mass (BMD) after implantation of a total hip stem can be precisely determined. However, prospective evaluation of the change of bone mass around the stem is only available for selected stems and short-term follow-up (up to 36 months). We analyzed BMD in patients who had undergone uncemented THA by DXA. Only patients with good clinical outcome (Merle d' Aubigne score > 12) were included to obtain normative data for regular bone response. Two separate studies were performed: a prospective longitudinal study over 84 months with baseline values acquired within the first postoperative week (group A) (n = 26 patients) and a separate cross-sectional study, median follow-up 156 (124-178) months (group B) (n = 35 patients). Regions of interest were defined according to Gruen (ROI 1-7) and as net average ROI (net avg) for the periprosthetic femoral bone. After the initial remodeling process (12 months), BMD was compared to the 84 month (longitudinal) and the 156-month (cross-sectional) follow-up values to determine long-term periprosthetic changes of bone mineral density. The longitudinal study (group A), after the initial bone remodeling, showed no relevant further bone loss for women and men with BMD values 1.19 +/- 0.15 and 1.40 +/- 0.19, respectively, 12 months (women 89.8%, men 93.6%), and 1.19 +/- 0.13 and 1.36 +/- 0.18, respectively, after 84 months (women 90.0%, men 91.3%) (P = 0.98, P = 0.08,) respectively. The distribution of the BMD around the stem changed during the first 12 months. The ROIs around the proximal stem (ROI 1 and 7) showed the lowest absolute values at the 12-month follow-up and BMD in ROI 7 decreased most during the further follow-up until 84 months. The cross-sectional study (group B) showed no significant difference in BMD (net avg) values at a median of 156 months follow-up compared to the 12-month values (group A) (women: P = 0.77, men: P = 0.44). Initial BMD, implant diameter, and body mass index did not influence BMD loss (net avg) in this study, whereas age showed a weak correlation with BMD loss. The results show that after the initial remodeling process, no relevant further bone loss (net avg) occurs up to 84 months postsurgery, and values after a median of 156 months are similar. Normative long term changes in the periprosthetic bone can be demonstrated in defined ROIs after implantation of a tapered corundum-blasted titanium stem with a good clinical result. PMID- 14565593 TI - A comparison study of the reference curves of bone mineral density at different skeletal sites in native Chinese, Japanese, and American Caucasian women. AB - To understand the differences among reference curves for bone mineral density (BMD) for Chinese, Japanese, and American Caucasian women, we measured the BMD at the anteroposterior (AP) lumbar spine (L1-L4), lateral lumbar spine (L2-L4), hip (including the femoral neck, trochanter, intertrochanter, Ward's triangle, and total hip), and ultradistal forearm by the dual-energy X-ray absorptiometry (DXA) in a total of 2728 healthy Chinese women, aged 5-96 years. Documented BMD data for Japanese women and device manufacturer's BMD new reference databases (including the NHANES III dataset) for American Caucasian women were also used in this study. The cubic regression model was found to fit best in analyzing the age associated variations of BMD at various sites in Chinese women, i.e., the equations had the largest coefficient of determination (R2). At the AP/Lat spine, trochanter, intertrochanter, and Ward's triangle, BMD reference curves for Chinese women were lower than those for Japanese or Caucasian women, while at the femoral neck, total hip, and ultradistal forearm, the reference curves for Chinese women were higher than those for Japanese women, with overlaps and crossing of the curves for some age spans in comparing the Chinese and Caucasian women. There were significant differences in the peak BMD (PBMD) at various sites among the Chinese, Japanese, and Caucasian women (P = 0.000). The PBMDs for Chinese women at the lumbar spine and various sites of the hip were 5.7% +/- 2.1% (mean +/- SD, range, 2.7-7.9%) lower than those for Japanese women and 5.1% +/- 2.7% (range, 0.5-7.2%) lower than those for Caucasian women; however, the PBMDs for Chinese women were 26.2% higher than those for Japanese women and 10% higher than those for Caucasian women at the ultradistal forearm. After the PBMD, average T-scores of Chinese women for losses at the AP lumbar spine with increasing age were nearly identical to those for Japanese women, but both were greater than those for Caucasian women. The average T-scores for BMD loss at various sites in Chinese women were higher than those for both Japanese and Caucasian women except at the femoral neck, where the T-scores of Chinese women were exceeded by those of both Japanese and Caucasian women. Estimated from the T score curve of BMD loss, the age of osteoporosis occurrence at the femoral neck in Chinese women was about 10 years later than that in Japanese or Caucasian women; at the AP spine, Chinese women were similar to Japanese women; at the other sites, the age for occurrence of osteoporosis in Chinese women was about 5 15 years earlier than that in either Japanese or Caucasian women. There are differences in prevalence or odds ratio (OR) of osteoporosis at the same skeletal region for Chinese, Japanese, and Caucasian women aged > or = 50 years or at different skeletal regions in women of the same race. The prevalences of osteoporosis at various regions of the hip in Chinese women are 10.1-19.8% and ORs are 22.0-32.3, of which prevalence at the femoral neck is the lowest (10.1%); the prevalences of osteoporosis in Japanese women are 11.6-16.8% and ORs are 21.1 26.3, of which prevalence at the femoral neck is the lowest (11.6%); and the prevalences of osteoporosis in Caucasian women are 13.0-20.0% and ORs are 19.4 48.9, of which prevalence at the femoral neck is the highest (20%). In conclusion, racial differences in BMD reference curves, prevalences, and risks of osteoporosis at various skeletal sites exist among native Chinese, Japanese, and American Caucasian women. PMID- 14565594 TI - Alendronate treatment in men with primary osteoporosis: a three-year longitudinal study. AB - Bisphosphonates have been widely used in the treatment of osteoporosis in women, whereas until now there have been few data on their use in men. The aim of this study was to evaluate the effect of a 3-year alendronate treatment on bone mineral density (BMD) and quantitative ultrasound (QUS) in men with primary osteoporosis. We studied 77 osteoporotic men (aged 57.1 +/- 10.8 yrs) who completed a 3-year treatment with alendronate (10 mg/day) plus calcium (1000 mg/day) (n = 39), or calcium alone (n = 38). At baseline and at a 12-month interval, we measured BMD at the lumbar spine and femur (femoral neck and total hip) by DXA (Hologic) and speed of sound (SOS), broadband ultrasound attenuation (BUA) and Stiffness (S) at the os calcis by Achilles plus (Lunar). Alendronate treatment had significantly increased lumbar spine BMD by 4.2% at year 1, by 6.3% at year 2, and 8.8% at year 3. BMD at the femoral neck and total hip had increased by 2.1% and 1.6% at year 1, by 3.2% and 2.9% at year 2, and by 4.2% and 3.9% at year 3, respectively. BUA and Stiffness showed a significant increase in the alendronate-treated group at year 2 (3.2% and 4.9%, respectively) and at year 3 (3.8% and 6%, respectively). BMD at the lumbar spine showed the best longitudinal sensitivity whereas longitudinal sensitivity of both QUS at the heel and femur BMD were similar. In conclusion, this study confirms that alendronate represents an important therapeutic advance in the management of male osteoporosis. BMD at the lumbar spine appears to be the best method for monitoring the effect of alendronate on bone mass in osteoporotic men. PMID- 14565595 TI - Association analysis of bone mineral density and single nucleotide polymorphisms in two candidate genes on chromosome 1p36. AB - Two candidate genes for bone mineral density (BMD), tumor necrosis factor alpha receptor 2 (TNFRSF1B) and lysyl hydroxylase (PLOD1), have been scanned for single nucleotide polymorphisms (SNPs) within their coding and promoter regions. These two genes, separated by about 200 kb, are located within the chromosomal interval 1p36.2-1p36.3 that has been linked to femoral neck BMD. In a patient population (n = 104) of European descent, there were four SNPs within TNFRSF1B and six SNPs within PLOD1 that occurred with greater than 5% frequency. There was significant linkage disequilibrium within both genes. Single marker analysis revealed significant association for one SNP located in intron 6 of PLOD1 and lumbar spine BMD (P = 0.01). Allelic haplotypes that encompassed the four SNPs in TNFRSF1B or the six SNPs in PLOD1 were assigned using a Bayesian algorithm as implemented in the program Haplotyper. Association of TNFRSF1B haplotypes with femoral neck BMD was statistically significant (P = 0.01). Similarly, PLOD1 haplotypes demonstrated a statistically significant association with spinal BMD (P = 0.04). These findings strengthen the potential importance of chromosome 1p36.2-1p36.3 in contributing to BMD variation, and are consistent with genetic variation in either PLOD1, TNFRSF1B or nearby genes playing a role in the phenotype. PMID- 14565596 TI - bFGF administration lowers the phosphate threshold for mineralization in bone marrow stromal cells. AB - Basic fibroblast growth factor (bFGF) is a potent mitogen and acts as an autocrine/paracrine factor for osteoblasts. Long-term administration of bFGF in vivo increases osteoblast number and stimulates matrix formation, but induces hypophosphatemia and impairs matrix mineralization. The goal of this study was to examine the interaction between bFGF and low levels of organic phosphate in an effort to better understand the possible long-term therapeutic effects of bFGF. These data show that in vitro administration of bFGF accelerates the calcification process and lowers the phosphate threshold needed for successful bone nodule formation. This correlates well with the observed upregulation of mRNA production for alkaline phosphatase and osteocalcin at day 7. These findings help elucidate the mechanisms of bFGF action on bone marrow stromal cell differentiation and mineralization and indicate that the delay in mineralization observed in vivo may not be caused by decreased phosphate availability alone. PMID- 14565597 TI - Regulated expression of G protein-coupled receptor kinases (GRK's) and beta arrestins in osteoblasts. AB - Desensitization of G-protein coupled receptors (GPCR's) is largely mediated by a family of enzymes and protein co-factors termed GRKs and arrestins, respectively. In the present studies, we investigated expression of GRKs and arrestins in osteoblastic cell lines concentrating on the enzymes (GRK2 and GRK3) and protein co-factors (beta-arrestint 1 and beta-arrestin 2) that play dominant roles in regulating GPCR responsiveness in most tissues and cell types. We found that osteoblastic cells express similar amounts of GRK2 with either undetectable or lesser amounts of GRK3. In contrast, expression of beta-arrestin 1 and beta arrestin 2 by osteoblastic cells varied between cell lines. To determine if GRK2 or beta-arrestin expression is modulated during osteoblast development, we assessed expression of GRK2 and beta-arrestin proteins during differentiation of the mouse osteoblastic cell line MC3T3-E1 cells over a 21-day period. We found that expression of GRK2 and beta-arrestin 2 increased to maximal levels by day 7 and then decreased 4-fold by day 21. In contrast, expression of beta-arrestin 1 increased to maximal levels by day 14 and then decreased 2-fold by day 21. Over this same time period (days 7-21), PTH/PTHrP receptor number decreased to a greater extent than the decrease in PTH(1-34)-induced cAMP generation, suggesting that responsiveness of individual PTH/PTHrP receptors was enhanced in differentiated cells. We conclude that (1) osteoblastic cell lines differentially express the enzymes and protein co-factors that modulate GPCR responsiveness and (2) expression of both GRK2 and beta-arrestins is temporally regulated during osteoblast development. These data are consistent with the notion that GPCR responsiveness may be differentially regulated in osteoblastic cell lines and during osteoblast development. PMID- 14565599 TI - Effects of phenol red on CFU-f differentiation and formation. AB - Osteoblastic induction is commonly studied using the colony-forming unit fibroblastic (CFU-f) assay, in which bone marrow stromal cells (BMC) are grown in a tissue culture environment permissive for osteoblastic differentiation (DMEM containing dexamethasone, ascorbic acid and beta-glycerophosphate). These cells form colonies, which express alkaline phosphatase, and form a collagenous matrix that becomes calcified. However, these same cells originate in the bone marrow where under normal circumstances they do not proliferate or differentiate despite being subjected to many of the same growth factors and hormones present within the tissue culture environment. We show here that phenol red, present within tissue culture medium as a pH indicator, may itself be a factor that permits osteoblastic recruitment. BMC cultured in the presence of the bone anabolic agents PGE2, PGA2, or bFGF, but in the absence of phenol red, failed to respond to these agents in terms of total or osteoblastic colony number. This effect was dose dependent, with low (2.5 mg/l) and high (15-20 mg/l) doses of phenol red being nonpermissive for the stimulatory effects of PGE2 whereas doses of 5-10 mg/l were permissive. Furthermore, the effects observed in the absence of phenol red could not be abrogated by the addition of 17beta-estradiol indicating that these effects cannot be attributed to estrogenic impurities within the phenol red preparation. This indicates that phenol red itself can affect the differentiation of BMC by a mechanism not previously described. PMID- 14565598 TI - Induction of apoptosis in skeletal tissues: phosphate-mediated chick chondrocyte apoptosis is calcium dependent. AB - In an earlier study, we have shown that Pi induced apoptosis of terminally differentiated hypertrophic chondrocytes. To ascertain whether Ca2+ modulates Pi induced cell death, we asked the following two questions: First, can we prevent Pi-induced apoptosis by removing Ca2+ from the culture medium; alternatively, can we potentiate cell death by increasing the Ca2+ concentration? Second, can we inhibit chondrocyte apoptosis by blocking Pi transport? We also explored the mechanism of apoptosis by evaluating mitochondrial activity and reactive oxygen species (ROS) generation in cells treated with the ion pair. We noted that EDTA and EGTA blocked Pi-induced apoptosis in a dose-dependent manner. While high levels of Ca2+ alone had little effect on chondrocyte viability, the cation enhanced Pi-dependent cell death and greatly increased Pi uptake. When Pi transport was blocked, there was complete inhibition of cell killing. The process of cell death was characterized by mitochondrial hyperpolarization; two hours following apoptogen treatment, there was a significant decrease in the mitochondrial membrane potential. Coincident with the changes in mitochondrial function, there was an increase in intracellular Ca2+ that was maintained throughout the experimental period. A raised Ca2+ signal was observed in blebs at the cell membrane. Finally, we noted that, 75 minutes after treatment with the ion pair, there was a six-fold elevation in ROS levels. This increase declined to baseline values after three hours. Based on these observations, we suggest that, at the cartilage mineralization front, an elevation in local environmental Ca2+ and Pi concentrations modulates oxidative metabolism, and triggers apoptosis of terminally differentiated chondrocytes. PMID- 14565600 TI - Increase in tartrate-resistant acid phosphatase of bone at the early stage of ascorbic acid deficiency in the ascorbate-requiring Osteogenic Disorder Shionogi (ODS) rat. AB - The effect of ascorbic acid deficiency on bone metabolism was evaluated using the ascorbate-requiring Osteogenic Disorder Shionogi (ODS) rat model. Ascorbic acid (Asc)-deficient rats gained body weight in a manner similar to Asc-supplemented rats (control) during 3 weeks, but began to lose weight during the 4th week of Asc deficiency. The tartrate-resistant acid phosphatase (TRAP) activity in serum increased to about 2-fold the control value in the rats fed the Asc-free diet for 2, 3, and 4 weeks (AscD2, AscD3, and AscD4), while a decrease in the alkaline phosphatase (ALP) activity was observed only in AscD4 rats. The serum pyridinoline cross-linked carboxyterminal telopeptide of type I collagen (ICTP) level significantly increased to 1.3-, 1.4-, and 1.9-fold of that in the controls in AscD2, D3, and D4, respectively. The ALP activity in the distal femur was unchanged in AscD1, D2, and D3, but decreased to 50% of the control level in AscD4 rats. The TRAP activity in the distal femur increased to about 2-fold of that in the controls in the AscD2 and D3 and decreased to the control level in the AscD4 rats. The amount of hydroxyproline in the distal femur significantly decreased to about 80%, 70%, and 60% of the control in AscD2, D3, and D4 rats, respectively. These decreases were associated with a similar reduction in the calcium content of the distal femur. Histochemical analysis of the distal femur showed an increase in TRAP-positive cells in AscD2 and AscD3 rats and a decrease in the trabecular bone in AscD2, D3, and D4 rats. These results suggested that a deficiency of Asc stimulated bone resorption at an early stage, followed by a decrease in bone formation in mature ODS rats which already had a well-developed collagen matrix and fully differentiated osteoblasts. PMID- 14565601 TI - Mineralization of basement membrane mediates dentogingival adhesion in mammalian and nonmammalian vertebrates. AB - In order to elucidate the mechanism of adhesion between the gingiva and the tooth, detailed comparative ultrastructural studies of the dentogingival border were done in the monkey and shark. The tissues were prepared with or without demineralization for the ultrastructural observations. At the border, the internal basement membrane, which is firmly bound to the junctional epithelium through hemidesmosomes, was specialized differently in these species. In the monkey, the lamina densa was closely associated at its enamel side with an additional layer which had characteristics of the lamina densa and was referred to as the supplementary lamina densa. In the shark, the lamina densa showed a unique, hemidesmosome-related specialization in the form of the intermittent occurrence of bulges along its surface facing the epithelium. In nondemineralized tissues a part of the basement membrane, that is, the supplementary lamina densa (monkey) and the main lamina densa but not bulges (shark), was preferentially mineralized. The mineral deposit was continuous with that in the enamel and enameloid/dentine, thus constituting an advancing edge of mineralization. The network arrangement of the mineral crystals in the monkey basement membrane resembled the pattern of the cord network of the basement membrane, suggesting the presence of a delicate mutual basement membrane-mineral interaction. Thus, the organic phase and the mineral phase are allowed to make contact at this mineralized area of the basement membrane and firmly bind to one another. Therefore, strong gingiva-tooth adhesion is established by partial mineralization of the internal basement membrane, in a way similar to that found in the previously reported association of maturation stage ameloblasts with the enamel. PMID- 14565602 TI - Effects of increased muscle mass on bone in male mice overexpressing IGF-I in skeletal muscles. AB - It has been hypothesized that increase in muscle mass increases the strain on bone resulting in increase in bone mass. The aim of the present study was to determine the effects of increased muscle mass on bone. A colony of transgenic mice that overexpress hIGF-I in muscle, resulting in larger muscles, was established. Six-month-old heterozygous and wild type males were used in this study. The tibial diaphysis, femoral diaphysis and distal femoral metaphysis were analyzed using pQCT densitometry. Heterozygous animals had significantly higher body weight, muscle weight and muscle area when compared with wild type animals. Tibia and femur of the heterozygous mice had significantly higher weights and lengths. The tibial and femoral diaphyses of heterozygous animals had significantly higher cortical bone area, cortical bone mineral content, cortical bone mineral density, cortical thickness and periosteal perimeter when compared with wild type animals. In the distal femoral metaphysis, the total bone area and the cancellous bone area of heterozygous mice were significantly higher than those of wild type animals. In conclusion, increased muscle mass was associated with bigger bones in animals overexpressing IGF-I. Only pure cortical bone increased in both area and mineral content in these animals; cancellous bone, however, increased only in area and not in mineral content and density. PMID- 14565603 TI - The evaluation of the HIV/AIDS drug access initiatives in Cote d'Ivoire, Senegal and Uganda: how access to antiretroviral treatment can become feasible in Africa. PMID- 14565604 TI - Virologic and immunologic outcomes and programmatic challenges of an antiretroviral treatment pilot project in Abidjan, Cote d'Ivoire. AB - BACKGROUND: In Cobte d'Ivoire, a pilot project was developed by UNAIDS and the Ministry of Health to improve access to AIDS care, including antiretroviral therapy, for adults and children infected with HIV. This evaluation of the project is the first to provide results of a large number of HIV-infected patients receiving antiretroviral therapy in West Africa. METHODS: We evaluated records of persons who presented for care from August 1998 to August 2000 at six accredited centers in Abidjan. Patients were treated with two nucleoside reverse transcriptase inhibitors (2NRTI) or highly active antiretroviral therapy (HAART). RESULTS: Of 2878 patients who were screened, 2351 (83%) were HIV-infected and eligible (CD4 T lymphocyte count < 500 x 10(6) cells/l or plasma HIV-RNA level > 10 000 copies/ml) for antiretroviral therapy. Of those who were eligible, 81% were symptomatic, 63% had a CD4 cell count < 200 x 10(6) cells/l, 12% had previously taken antiretroviral drugs, and 56% returned to the clinic for follow up. Of the patients screened, 768 (27%) were started on antiretroviral therapy, including 450 on HAART, 296 on 2NRTI, and 22 on other regimens. We analyzed data from 480 HIV-1-infected adults, who were naive to therapy, were prescribed HAART or 2NRTI, and had at least one clinic visit after starting therapy. In an intent to-treat analysis of patients who received HAART, the estimated plasma HIV-1 RNA level was approximately 1.9 log10 copies/ml (80-fold) lower, while estimated CD4 cell count was > 100 x 10(6) cells/l higher than baseline values, after 1 year of therapy. Approximately 25% of adults on 2NRTI and 50% of those on HAART had < 200 copies/ml, after 1 year of therapy. The probability of an adverse event occurring within 6 months after starting therapy was 0.20. The probability of survival for at least 1 year was 0.84 (95% confidence interval, 0.80-0.89). CONCLUSION: After starting antiretroviral therapy, these HIV-1-infected patients in West Africa had similar virologic and immunologic outcomes, probability of an adverse event, and estimated survival, as patients enrolled in clinical trials in the USA and Europe. However, only one-third of eligible patients received therapy, highlighting the importance of providing adequate education and support for initiating and adhering to therapy in this and similar programmes. PMID- 14565605 TI - Changes in levels of immune activation and reconstitution markers among HIV-1 infected Africans receiving antiretroviral therapy. AB - OBJECTIVE: To describe changes in immune activation and reconstitution markers among HIV-1-infected patients receiving antiretroviral therapy (ART) in Abidjan, Cote d'Ivoire. METHODS: Between November 1998 and February 2001, we analyzed changes in immune activation and reconstitution markers among 52 patients. Good virologic responders (n = 26) were defined as those who had suppressed and maintained plasma viral load (VL) below the detection limit of the assay for at least 12 months. Poor virologic responders (n = 26) were defined as those with a detectable VL at 6 and 12 months after beginning ART. RESULTS: Of the 26 good virologic responders, 20 (77%) were on highly active antiretroviral therapy (HAART) compared with one (4%) of the poor responders. Among the 26 good responders, baseline median levels of CD38+CD8+ T cells were elevated, but had decreased significantly at 6 months (P < 0.001) and at 12 months of therapy (P < 0.001). Median levels of HLA-DR+CD8+ T cells also decreased from baseline at 6 months (P < 0.001) and at 12 months of therapy (P < 0.001). Levels of CD62L+CD4+ T cells increased steadily during the 6 and 12 months of therapy and reached levels observed among HIV-negative blood donors (P = 0.07). Among the 26 poor responders, median levels of CD38+CD8+ T cells decreased significantly at 12 months of therapy (P = 0.006), but were higher than levels in blood donors (P = 0.005). Levels of HLA-DR+CD8+ T cells decreased significantly at 12 months of therapy (P < 0.001). Levels of CD62L+CD4+ decreased over time. CONCLUSION: Our results suggest that HAART can be successfully used in African populations with elevated baseline immune activation markers. PMID- 14565606 TI - Prevalence of genotypic and phenotypic HIV-1 drug-resistant strains among patients who have rebound in viral load while receiving antiretroviral therapy in the UNAIDS-Drug Access Initiative in Abidjan, Cote d'Ivoire. AB - OBJECTIVE: To determine the prevalence of genotypic and phenotypic antiretroviral (ARV) drug-resistant HIV-1 strains among patients with viral load rebound while receiving ARV therapy in Abidjan, Cote d'Ivoire. METHODS: Between August 1998 and April 2000, we selected all patients (n = 241) who had received ARV drug therapy for at least 6 months in the UNAIDS-Drug Access Initiative (DAI), in Abidjan. We analyzed for genotypic and phenotypic drug resistance among 97 (40%) of the 241 patients who had a rebound in plasma viral load, defined as an initial decrease of > 0.5 log10 copies/ml followed by a subsequent increase of > 0.25 log10 copies/ml. RESULTS: Of the viruses isolated from the 97 patients, 86 (88.7%) had usable sequences and 68 (79%) of the 86 patients had genotypic resistance to at least one reverse transcriptase inhibitor (RTI) or protease inhibitor (PI). Resistant mutations were found for zidovudine in 50 (78%) of 64 patients who had received the drug, 11 (68.7%) of 16 patients on lamivudine, for nevirapine in two (2%), for indinavir in one (1%), and for ritonavir in one (1%). Phenotypic resistance to at least one nucleoside RTI was seen in 45 (56%) of the 80 patients tested, to non-nucleoside RTIs in eight (10%), and to PIs in one (1.3%). Multivariate regression analysis showed factors associated with resistance to be initial treatment with dual therapy (P = 0.04) compared with highly active antiretroviral therapy, and maximal initial viral load response (P = 0.006). CONCLUSION: Our results demonstrate a high prevalence of ARV drug resistance associated with dual ARV therapy. These results indicate the limited role for dual ARV therapy. PMID- 14565607 TI - Low rate of genotypic HIV-1 drug-resistant strains in the Senegalese government initiative of access to antiretroviral therapy. AB - OBJECTIVE: To monitor the prevalence of antiretroviral (ARV)-resistant HIV-1 viruses, and the genotypic mutations in patients enrolled in the Senegalese initiative for access to antiretroviral treatment (ART). METHODS: A total of 80 patients with a virological follow-up of at least 6 months were selected, 68 were ART-naive and 12 ART-experienced. Genotypic resistance to ARV was studied at baseline for a random subset of patients and at each rebound in plasma viral load during ART, by sequencing the protease and reverse transcriptase genes. RESULTS: At baseline, 66 patients received highly active antiretroviral therapy (HAART) [2 nucleoside reverse transcriptase inhibitors (NRTIs) +1 protease inhibitor (PI) (n = 64) or 2 NRTIs + 1 non-nucleoside reverse transcriptase inhibitor (NNRTI) (n = 2)] and 14 patients (17.5%) started with a dual therapy because of ongoing antitubercular therapy or efficient previous bitherapy for the ART-experienced patients. The emergence of drug-resistant viruses (n = 13) during follow-up was more frequent in ART-experienced patients than in ART-naive patients, 41.7 versus 11.8%, resistant viruses emerged at comparable follow-up periods, a median of 17.8 and 18.3 months, respectively. In patients receiving zidovudine and lamivudine in their drug regimen, resistance to lamivudine was more frequent than to zidovudine. Two of the three patients, with viruses resistant to PIs, acquired mutations associated with cross-resistance. Strikingly, five (39%) of the 13 patients developed resistances to drugs that they had never received (n = 3) or that they received 18 or 36 months ago (n = 2). Didanosine/stavudine pressure had selected zidovudine-resistant viruses in four patients, and indinavir had selected a nelfinavir-resistant virus in one patient. CONCLUSION: In contrast to other reports from developing countries where patients had received ARVs in an uncontrolled manner, our study showed that implementation of HAART together with good clinical, biological and logistical monitoring can reduce the emergence of resistant strains in Africa. PMID- 14565608 TI - Development of phenotypic and genotypic resistance to antiretroviral therapy in the UNAIDS HIV Drug Access Initiative--Uganda. AB - OBJECTIVE: We describe phenotypic drug resistance, response to therapy, and genotypic mutations among HIV-infected patients in Uganda taking antiretroviral medications for > or = 90 days who had a viral load > or = 1000 copies/ml. METHODS: HIV-1 group and subtype, virologic and immunologic responses to antiretroviral therapy, phenotypic resistance to antiretroviral drugs, and associated genotypic mutations among patients at three treatment centers in Uganda between June 1999 and August 2000 were assessed. Therapy was two nucleoside reverse transcriptase inhibitors (NRTIs) or highly active antiretroviral therapy (HAART). RESULTS: All HIV identified was HIV-1, group M, subtypes A, C, and D. Sixty-one (65%) of 94 patients with a phenotypic resistance result had evidence of phenotypic resistance including resistance to a NRTI for 51 of 92 (55%) taking NRTIs, to a non-nucleoside reverse transcriptase inhibitor (NNRTI) for nine of 16 (56%) taking NNRTIs, and to a protease inhibitor (PI) for eight of 37 (22%) taking PIs. At the time of the first specimen with resistance, the median change from baseline viral load was -0.56 log copies/ml [interquartile range (IQR), -1.47 to +0.29] and CD4+ cell count was +35 x 10(6) cells/l (IQR, 18 to +87). Genotypic resistance mutations, matched with phenotypic resistance assay results and drug history, were generally consistent with those seen for HIV 1, group M, subtype B infections in industrialized countries. CONCLUSION: Initial phenotypic resistance and corresponding genotypic mutations among patients treated in Uganda were similar to those with subtype B infections in North America and Europe. These data support policies that promote the use of HAART regimens against HIV-1, group M, non-B subtypes in a manner consistent with that used for subtype B infections. PMID- 14565609 TI - Antiretroviral therapy in HIV-2-infected patients: changes in plasma viral load, CD4+ cell counts, and drug resistance profiles of patients treated in Abidjan, Cote d'Ivoire. AB - OBJECTIVE: To describe changes in plasma viral load, CD4+ cell counts, and drug resistance profiles of HIV-2-infected patients receiving antiretroviral (ARV) therapy in Abidjan, Cote d'Ivoire. METHODS: Consecutive blood samples were collected from 18 HIV-2-infected ARV-naive patients who had received ARV therapy in the UNAIDS drug access initiative (UNAIDS-DAI) in Abidjan between August 1998 and July 2000. Changes in HIV-2 plasma viral load, CD4+ cell counts, and genotypic and phenotypic drug resistance testing were determined. RESULTS: At baseline, 11 (61%) of the 18 patients initiated highly active antiretroviral therapy (HAART) and seven (39%) received dual therapy. No significant change in median viral load was observed at 2 months (P = 0.09), at 6 months (P = 0.06), and at 12 months of therapy (P = 0.26). No significant increase in CD4+ cell counts was observed at 12 months (P = 0.10). All four patients on indinavir containing HAART had undetectable viral loads at 2-4 months of therapy. However, none of seven patients on nelfinavir-containing HAART had a substantial decrease in viral load. Viruses from 14 patients were analyzed, 12 of which (86%) had at least one primary resistance mutation that is known to confer resistance to HIV-1 virus. Three patients had the multi-drug-resistant mutation, Q151M, two of whom showed reduced susceptibility to zidovudine, didanosine, stavudine and zalcitabine. CONCLUSION: Our limited findings show that nelfinavir-containing regimens may have limited virologic benefit to HIV-2-infected patients. PMID- 14565610 TI - Clinical, immunological and virological response to different antiretroviral regimens in a cohort of HIV-2-infected patients. AB - OBJECTIVE: To assess the clinical, immunological and virological response and the emergence of resistance towards antiretroviral therapy (ART) in a cohort of HIV-2 infected patients. DESIGN: Observational study. PATIENTS: HIV-2-infected patients residing in the Netherlands. RESULTS: From 1995 to 2001 seven patients failed various ART regimens. The resistance mutations were analysed retrospectively. Development of mutations proved to be similar to that observed in HIV-1-infected patients, with the exception of a higher occurrence of the Q151M mutation within the reverse transcriptase gene. In a prospective study, comprising 13 consecutive naive HIV-2-infected patients, all patients achieved plasma HIV-2-RNA suppression below the detection limit (500 copies/ml). The antiretroviral regimen consisted of two nucleoside reverse transcriptase inhibitors (NRTIs) and indinavir, with a boosting dose of ritonavir; the median follow-up was 91 weeks. Two patients experienced a temporary virological rebound, while at the same time therapeutic drug monitoring showed sub-therapeutic plasma levels of indinavir. CONCLUSION: Sustained viral suppression in HIV-2-infected patients can be achieved using an antiretroviral regimen of two NRTIs and boosted indinavir or lopinavir. PMID- 14565611 TI - Socio-economic and health characteristics of HIV-infected patients seeking care in relation to access to the Drug Access Initiative and to antiretroviral treatment in Cote d'Ivoire. AB - OBJECTIVE: To compare socio-economic and health characteristics of HIV-infected patients in Cote d'Ivoire whether or not they had access to the Drug Access Initiative (DAI) and to antiretroviral drug (ARV) treatment. DESIGN AND METHODS: Cross-sectional survey using medical files, blood sampling for CD4 cell counts and face-to-face interviews among all patients, informed of their HIV status, who attended during a 6-week period in the five DAI referral centres and three additional centres in charge of HIV care in Abidjan and Bouake (participation rate = 65.4%). Multiple logistic regression using generalized estimating equations (GEE) to identify factors related to non-access to DAI and to ARV treatment. RESULTS: Among the 711 respondents, 23.0% were ARV-treated, 14.2% had been included in the DAI but were still waiting for initiation of ARV, and 62.7% were neither part of the DAI nor ARV-treated. In this latter group, less than one third (29.6%) declared that they knew about the existence of the DAI. Among the 164 ARV-treated patients, 59.1% had benefited from DAI public subsidies partially covering the costs of drugs. In the non-DAI-non-ARV-treated group, 86% could have qualified for ARV treatment according to the DAI medical criteria (CD4 cell counts < 500 x 10(6) cells/l), and only 32.9% of those medically eligible were prescribed cotrimoxazole prophylaxis. In multivariate analysis, not being in the DAI and not being ARV-treated was related to: being a male, not having health care insurance, having a low level of education, living in poor housing conditions (absence of refrigerator in the household, absence of ventilation in patient's bedroom), and not being under cotrimoxazole prophylaxis. CONCLUSION: The Ivoirian DAI has facilitated access to ARV treatment for a significant number of patients with limited ability to pay. The majority of HIV-infected patients seeking care however face persisting socio-economic and informational barriers to access to these treatments. PMID- 14565612 TI - Access to antiretroviral treatment and sexual behaviours of HIV-infected patients aware of their serostatus in Cote d'Ivoire. AB - OBJECTIVE: To compare sexual behaviours of HIV-infected patients in Cote d'Ivoire whether or not they had access to antiretroviral treatment (ARV). DESIGN AND METHODS: Cross-sectional survey using face-to-face interviews among all patients, informed of their HIV serostatus, attending the eight health centers in charge of HIV care in Abidjan and Bouake. Univariate comparisons of declared sexual behaviours during the prior 6 months between the 164 ARV-treated and the 547 non ARV-treated respondents. Multiple logistic regressions to identify factors related to sexual abstinence in the total sample and to unprotected sexual intercourse in the subsample of sexually active respondents during the same period were performed. RESULTS: More than half of the 711 respondents (53.0%) declared an absence of sexual activity during the previous 6 months, with this proportion being significantly higher among women (60.6%), and among both men (85.7%) and women (92.4%) who were not living in a stable relationship. Among the 334 sexually active patients, 49.7% declared a low frequency of sexual intercourse ('once a month or less'), and 43.7% declared at least one episode of unprotected sexual intercourse. In multivariate analysis, recent diagnosis of HIV infection (< 9 months), having only one sexual partner and not knowing her/his serostatus, high alcohol consumption, absence of episodes of acute morbidity, not participating in household's expenditures and not being ARV-treated were significantly related to a higher likelihood of HIV-related risky sexual behaviours. CONCLUSION: Sexual abstinence is the preventive strategy of choice for a majority of HIV-infected patients aware of their serostatus and consulting for care in Cote d'Ivoire. In these patients, access to ARV is not associated with an increase in HIV-related risky sexual behaviours. PMID- 14565613 TI - Physicians' knowledge and attitudes toward HIV care in the context of the UNAIDS/Ministry of Health Drug Access Initiative in Cote d'Ivoire. AB - OBJECTIVE: To evaluate the impact of the availabity of highly active antiretroviral therapies (HAART) in the context of the Drug Access Initiative (DAI) on physicians' knowledge, attitudes and practices toward HIV care in Cote d'Ivoire. DESIGN AND METHODS: Cross-sectional survey using self-administered questionnaires among all consulting physicians in the six 'referral centers' of the DAI and five additional centers in charge of HIV care. RESULTS: Among the 123 respondents (response rate = 82.0%), 45.1% took care of more than 20 HIV-infected patients during the previous year. These physicians with the most experience in HIV care had a better knowledge than the rest of the sample about HIV disease, cotrimoxazole prophylaxis and antiretroviral treatment, and were more likely to declare that HIV-infected patients may be 'dangerous for others' (33.9 versus 17.9%; P = 0.03). Although 54.5% declared that the eligibility medical criteria for HAART 'should be the same in both developing and developed countries', only 30.9% adhered to the recently issued DAI guideline (October 1999) recommending initiation of HAART for patients with CD4 cell counts < 500 x 10(6) cells/l. CONCLUSION: Physicians involved in the DAI in Cote d'Ivoire have acquired appropriate expertise and knowledge about HAART, but dissemination of information about HAART must be extended to physicians with more limited experience in HIV care. Current international efforts to adapt HIV treatment guidelines for resource-limited settings may face difficulties for reaching consensus among the African health professionals in charge. PMID- 14565614 TI - The most efficient use of resources to identify those in need of antiretroviral treatment in Africa: empirical data from Cote d'Ivoire's Drug Access Initiative. AB - OBJECTIVE: To describe the cost and outcome associated with the use of CD4 cell count and viral load tests as part of screening strategies to identify persons eligible for subsidized antiretroviral therapy (ART) in Cote d'Ivoire. METHODS: Empirical data from the Drug Access Initiative in Cote d'Ivoire (DAI-CI) were used to describe the laboratory cost of patient screening using sequential clinical staging, CD4 cell count, and viral load and the proportion of screened patients identified as eligible for ART. We also estimated costs modelling a parallel screening algorithm, across a range of laboratory costs and with current international recommendations to assess treatment eligibility. Benefit was defined as being found eligible for ART. RESULTS: Of the 2138 HIV-positive, ART naive, adults who presented to the DAI-CI between July 1998 and July 2000, median CD4 cell count was 172 x 10(6) cells/microl. DAI-CI criteria identified 2057 (96%) of these persons eligible for antiretroviral treatment. In a serial screening algorithm, 75% were eligible by CDC clinical stage B or C; 18% by CD4 cell count less than 500 x 10(6) cells/microl; and an estimated 3.9% by a viral load greater than 10 000 copies/ml. Use of the current US recommendations and a serial algorithm would have resulted in 1977 (92%) persons eligible for ART: 75% by CDC clinical stage B or C; 15% by CD4 cell count less than 350 x 10(6) cells/microl (including 8% < 200 x 10(6) cells/microl); and an estimated 3.6% due to viral load greater than 55 000 copies/ml. Using DAI-CI criteria and heavily subsidized laboratory test costs, the addition of CD4 cell count to clinical criteria cost US dollar 50 (serial algorithm) and US dollar 203 (parallel algorithm) to identify each additional eligible person. Modelling current recommendations with a serial algorithm, CD4 cell count cost an average US dollar 62/eligible person (US recommendations) and US dollar 109 (WHO recommendations). The addition of viral load cost between US dollar 108 (serial algorithm DAI) to US dollar 1700 (parallel algorithm DAI) to identify each additional eligible person. CONCLUSION: In the African context of scarce resources and the huge unmet demands for voluntary HIV testing and for ART, simple screening strategies are needed to identify those most in need of ART. Health personnel should be trained to identify and refer clinically symptomatic persons. Viral load testing is of high cost and dubious benefit and should not be part of screening algorithms for initiating ART. PMID- 14565615 TI - Access to antiretroviral drugs and AIDS management in Senegal. AB - OBJECTIVES: Description and analysis of the Senegalese Antiretroviral Drug Access Initiative (ISAARV), the first governmental highly active antiretroviral therapy (HAART) treatment programme in Africa, launched in 1998. METHODS AND RESULTS: ISAARV was initially an experimental project designed to evaluate the feasibility, efficacy and acceptability of HAART in an African context. It was based on four principles: collective definition of the strategy, with involvement of the health professionals who would be called on to execute the programme; matching the objectives to available means (gradual enrollment according to drug availability); monitoring by several research programmes; and ongoing adaptation of treatment and follow-up according to the latest international recommendations. Persons qualifying for antiretroviral (ARV) therapy are selected on the basis of immunological and clinical criteria, regardless of economic and social considerations. A system of subsidies was created to favor access to ARV. Following the ARV price reductions that occurred in November 2000, 100% subsidies were created for the poorest participants. Optimal adherence was ensured by monthly follow-up by pharmacists and support groups held by social workers and patient associations. The chosen supply and distribution system allowed drug dispensing to be strictly controlled. CONCLUSION: The ISAARV programme demonstrates that HAART can be successfully prescribed in Africa. This experience has served as the basis for the creation of a national treatment programme in Senegal planned to treat 7000 patients by 2006. PMID- 14565616 TI - Adherence to HAART and its principal determinants in a cohort of Senegalese adults. AB - BACKGROUND: Access to programmes providing highly active antiretroviral therapy (HAART) is recent in Africa. In Senegal, a national initiative was launched in 1998. The capacity of African patients to adhere to complex antiretroviral treatments (ARV) is largely unknown. METHODS: We assessed adherence and identified the main reasons for treatment interruption in a prospective observational cohort of patients participating in an ARV access programme in Dakar, Senegal. Adherence was estimated each month on the basis of the patients' stated consumption and on the proportion of the prescribed dose returned unused to the dispensing pharmacy. A total of 158 patients were studied between November 1999 and October 2001. RESULTS: A cross-section analysis showed that the stated level of adherence was high: on average, over the study period, the patients said they had taken 91% of each monthly dose and that they had taken the full monthly dose during two-thirds of the months studied. Adherence tended to be better among patients who were required to make little or no contribution to the cost of their treatment, through an appropriate pricing structure. Adherence was also better with efavirenz-containing regimens than with indinavir-containing regimens. CONCLUSION: These results show that adherence to HAART can be as high in Africa as that generally observed in industrialized countries, and that the cost and type of drug regimen must be taken into account when designing ARV access programmes for poor communities. PMID- 14565617 TI - Antiretroviral use in Ouagadougou, Burkina Faso. AB - Burkina Faso has the second highest seroprevalence rate for HIV in West Africa, estimated at 6.5% of the population. Although it is one of the poorest countries in the world, antiretrovirals have been used on an extremely limited basis in Burkina Faso since at least the early 1990s. In this article we will review the evolution of antiretroviral availability in this country, describe the mechanisms by which drugs are being accessed, and review our experience with expanding antiretroviral access through drug donations in community-based settings. Finally, we will discuss some of the implications for future attempts to expand access to treatment for people living with HIV in Africa. PMID- 14565618 TI - Antisense modulation of the coding or regulatory sequence of the folate receptor (folate binding protein-1) in mouse embryos leads to neural tube defects. AB - BACKGROUND: Although folic acid decreases the incidence of neural tube defects (NTDs) in humans, the mechanism for this protection is unknown. We have employed antisense technology to alter expression of the gene for the folate receptor (folate binding protein-1 [Folbp1]) in mouse embryos cultured in vitro. METHODS: Embryos were explanted on day 8 of gestation and cultured for 44 hr. Several oligodeoxyribonucleotides designed to modulate the coding region or a regulatory sequence in the 5'-untranslated region of Folbp1 were microinjected into the amniotic sac of embryos at the beginning of the culture period. RESULTS: Two different antisense sequences to the 5' and 3' coding region in Folbp1 produced concentration-dependent increases in the number of embryos with NTDs. Coinjection of 5-methyltetrahydrofolate with these sequences decreased the frequency of abnormal embryos. A semi-quantitative RT-PCR technique used to measure the amount of Folbp1 mRNA in treated and control embryos confirmed that the mRNA level was decreased by treatment with the antisense sequences. An antisense oligodeoxyribonucleotide to a 17 base cis regulatory element also generated a concentration-dependent increase in the frequency of embryos with NTDs, and a decrease in the level of Folbp1 mRNA. CONCLUSIONS: These results demonstrate that alterations in expression of Folbp1 by perturbing either the coding sequence or a critical regulatory cis-element can play a role in NTDs. PMID- 14565619 TI - Trichloroethylene effects on gene expression during cardiac development. AB - BACKGROUND: Halogenated hydrocarbon exposure is associated with changes in gene expression in adult and embryonic tissue. Our study was undertaken to identify differentially expressed mRNA transcripts in embryonic hearts from Sprague-Dawley rats exposed to trichloroethylene (TCE) or potential bio-transformation products dichloroethylene (DCE) and trichloroacetic acid (TCAA). METHODS: cDNA subtractive hybridization was used to selectively amplify expressed mRNA obtained from control or halogenated hydrocarbon exposed rat embryos. The doses used were 1100 and 110 ppm (8300 and 830 microM) TCE, 110 and 11 ppm (1100 and 110 microM) DCE, and 27.3 and 2.75 mg/ml (100 and 10 mM) TCAA. Control animals were given distilled drinking water throughout the period of experiments. RESULTS: Sequencing of over 100 clones derived from halogenated hydrocarbon exposed groups resulted in identification of numerous differentially regulated gene sequences. Up-regulated transcripts identified include genes associated with stress response (Hsp 70) and homeostasis (several ribosomal proteins). Down-regulated transcripts include extracellular matrix components (GPI-p137 and vimentin) and Ca(2+) responsive proteins (Serca-2 Ca(2+)-ATPase and beta-catenin). Two possible markers for fetal TCE exposure were identified: Serca-2 Ca(2+)-ATPase and GPI p137, a GPI-linked protein of unknown function. Differential regulation of expression of both markers by TCE was confirmed by dot blot analysis and semi quantitative RT-PCR with levels of TCE exposure between 100 and 250 ppb (0.76 and 1.9 microM) sufficient to decrease expression. CONCLUSIONS: Sequences down regulated with TCE exposure appear to be those associated with cellular housekeeping, cell adhesion, and developmental processes, while TCE exposure up regulates expression of numerous stress response and homeostatic genes. PMID- 14565620 TI - Vascular endothelial growth factor rescues 2,3,7,8-tetrachlorodibenzo-p-dioxin inhibition of coronary vasculogenesis. AB - BACKGROUND: We previously demonstrated that the environmental pollutant 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) reduces coronary vascular development in chick embryos in vivo. In the current study, we assessed whether TCDD inhibits early events in coronary endothelial tube formation and outgrowth, and whether this inhibition occurs through a vascular endothelial growth factor (VEGF)-dependent mechanism. METHODS: Fertile chicken eggs were treated with control (corn oil) or TCDD (0.3 pmol TCDD/g) on incubation day 0. On embryonic day 6, cardiac ventricle explants were cultured on a three-dimensional collagen gel, when coronary angioblasts are present, but prior to their assembly into endothelial tubes. Endothelial cells migrating out from explants were identified by immunohistochemistry, and endothelial tube number and length were quantitated. In addition, on incubation days 6 and 8, cardiac VEGF mRNA and protein were measured by reverse transcriptase-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. RESULTS: Endothelial tube length and number were significantly reduced (40% +/- 1.7% and 36% +/- 3%, respectively) in TCDD explants, compared to controls. Recombinant exogenous VEGF, as well as hypoxic stimulation with CoCl2 or 10% O2, significantly increased the length and number of outgrowing tubes in TCDD cultures, and this stimulation was prevented by a VEGF neutralizing antibody. In contrast, VEGF neutralizing antibody reduced the length and number of tubes only in control cultures, and had no inhibitory effect on tube outgrowth from TCDD explants. Finally, hearts from TCDD-treated embryos exhibited a significant reduction in both VEGF mRNA and protein, compared to controls. CONCLUSIONS: These data suggest that TCDD inhibits early coronary vascular outgrowth via a VEGF-dependent mechanism. PMID- 14565621 TI - Diarrhea: a new risk factor for neural tube defects? AB - BACKGROUND: Neural tube defects (NTDs) affect approximately 4000 US pregnancies annually. Folic acid supplementation taken before conception protects against the occurrence of NTDs. Adequate levels of vitamin B12 also appear to play a significant role. Gastrointestinal disturbances, such as those caused by diarrhea, might negatively affect the availability of these vitamins, thereby increasing the risk of these birth defects. METHODS: To determine whether periconceptional diarrhea increases the risk of NTD-affected pregnancies, a population-based case-control study was conducted in the 14 Texas-Mexico border counties. Information on diarrhea and other risk factors was ascertained by in person interview. Study subjects were Mexican-American women who resided and delivered in any border county during 1995-2000. Case women, identified through active surveillance, had liveborn or stillborn infants or fetuses diagnosed with anencephalus, spina bifida, or encephalocele. Control women were randomly selected from women delivering normal live births in study area health facilities. RESULTS: One or more episodes of periconceptional diarrhea were associated with increased risk of NTD-affected pregnancies compared to no episodes of diarrhea (OR = 3.7, 95% CI = 1.8-7.6). This association was independent of fever, obesity, maternal age, maternal birthplace, income, prior unproductive pregnancy, and dietary plus multivitamin folate intake, known risk factors for NTDs. CONCLUSIONS: Confirmation of this new risk factor might have public health implications due to the feasibility of modifying exposure. PMID- 14565622 TI - Low maternal alcohol consumption during pregnancy and oral clefts in offspring: the Slone Birth Defects Study. AB - BACKGROUND: The association between maternal alcohol consumption during pregnancy and oral clefts in offspring remains unclear. We studied this relation in a case control surveillance study of birth defects. METHODS: From 1983 to 1997, we recruited 5956 study subjects from greater Boston, Philadelphia, Toronto, and parts of Iowa. The cases were liveborn infants with cleft palate alone (CP; n = 205), cleft lip and palate (CLP; n = 383), cleft lip alone (CL; n = 259), or Pierre-Robin sequence (n = 65). The controls (n = 4272) were infants who had no oral clefts but had one or more of the following defects: malformations of the digestive tract, reproductive organs, abdominal wall, and respiratory tract; chromosomal anomalies; inguinal hernia; tumors; and Mendelian inherited disorders. Based on maternal reports of alcohol consumption during the first 4 months of pregnancy, we derived average weekly consumption, average number of drinks per drinking day, and the maximum number of drinks consumed in a given day. The mothers also provided data on potential confounding or modifying variables, such as vitamin supplement use. RESULTS: There was no relation between maternal alcohol consumption during pregnancy and CL or CP. The odds ratios (ORs) for cleft lip with or without palate (CL/P) were 1.0, 1.1, and 0.9 in women who consumed <1.0, 1.0-2.9, and 3.0 + drinks per week, respectively. These findings did not change when we considered possible modifying effects of vitamin supplement use. CONCLUSIONS: Our findings do not support an association between oral clefts and a low level of alcohol consumption. PMID- 14565623 TI - Birth defects in the offspring of non-Caucasian, non-Indigenous women in Western Australia. AB - BACKGROUND: Birth defects in infants born to non-Caucasian, non-Indigenous mothers in Australia have not been described in detail previously. METHODS: Using data from the Western Australian Maternal and Child Health Research Data Base, an index cohort of all non-Caucasian, non-Indigenous mothers giving birth in Western Australia between 1985 and 1995 (n = 17,706 births) was compared with a 10% random sample of births to Caucasian mothers (n = 24,180) over the same period. RESULTS: Overall, the index mothers had a 21% less risk of having a baby with birth defects (prevalence ratio, 0.79; 95% confidence interval, 0.72-0.86). The six major countries of birth of the index mothers were Vietnam, the Philippines, Singapore, Malaysia, India, and New Zealand. Mothers from Vietnam and the Philippines were significantly less likely to have a baby with a birth defect. The prevalences for the remaining four maternal countries of birth were not significantly different from that in the comparison cohort. CONCLUSIONS: The difference in risk of birth defects is probably not a result of confounding by maternal age, plurality, parity, socioeconomic status, marital status, or place of residence. However, it may be a real difference or a result of ascertainment bias. PMID- 14565624 TI - Socioeconomic effects on the risk of having a recognized pregnancy with Down syndrome. AB - BACKGROUND: Appoximately 95% of Down syndrome (DS) cases are caused by an error in germ cell division (meiosis), resulting in an extra chromosome 21. The meiotic error, predominantly of maternal origin, occurs either during the mother's fetal life (meiosis I) or at ovulation (meiosis II). Because maternal-age-specific DS prevalence rates vary between and within populations, it has been hypothesized that environmental factors can affect the risk for a DS pregnancy. METHODS: In a population-based case-control study of 997 clinically recognized DS cases (including fetal losses) and 1007 controls without a birth defect, we examined the mother's socioeconomic status (SES) from the time of her fetal life to the time of conception. SES variables were considered as proxies for environmental factors. We used multiple logistic regression for the analyses. RESULTS: We found associations with low levels of each SES variable examined: mother's education less than high school (OR, 1.29; 95% CI, 1.01-1.65), father's low occupation (OR, 1.23; 95% CI, 0.95-1.60), father's low education (OR, 1.28; 95% CI, 0.99-1.64), mother's father's low occupation (OR, 1.35; 95% CI, 1.06-1.71), and family income <$20,000 (OR, 1.31; 95% CI, 1.02-1.68) [corrected]. The risk for DS increased as the number of low socioeconomic factors present throughout the mother's life increased. With four factors present, the risk (adjusted for confounders) almost doubled (OR, 1.98; 95% CI, 1.30-3.01). Those associations persisted among young (< 30) and old (> or = 30) maternal age groups. CONCLUSIONS: A mother's low SES during any period before conception increases her risk for a recognized pregnancy with DS. Because of the high birth prevalence of DS, the public health impact of maternal SES may be considerable. PMID- 14565625 TI - Frequency of neural tube defects in Mexico. PMID- 14565626 TI - Temporal aspects of changes in neural tube defect rates. PMID- 14565627 TI - Influence of smoking on predictors of vascular disease. AB - Beyond the already well-established strong causative relationship with cancer, smoking increases the risk for vascular disease. Smoking may act directly or adversely influence risk factors contributing to the development of vascular disease. Smoking causes endothelial dysfunction, dyslipidemia (decreased high density lipoprotein cholesterol levels, hypertriglyceridemia and increased oxidation of low-density lipoprotein cholesterol) and platelet activation leading to a prothrombotic state. Smoking increases emerging risk factors (eg, fibrinogen, homocysteine, and high-sensitivity C-reactive protein) and increases insulin resistance and the risk of developing type 2 diabetes mellitus. The beneficial effects of statins and antioxidants (eg, vitamins C and E, beta carotene) are counteracted by smoking. Smoking-induced alterations in growth factors, adhesion molecules, and even in genes can accelerate the progression of atherosclerosis. The aim of this review is to consider the adverse consequences of smoking on the factors predisposing to vascular disease and to emphasize the beneficial effects of smoking cessation. PMID- 14565628 TI - Prevention of venous thrombosis in long-haul flights with Flite Tabs: the LONFLIT FLITE randomized, controlled trial. AB - The aim of this study was to evaluate the development of edema, and superficial and deep vein thrombosis (DVT) prophylaxis with an oral profibrinolytic agent (Flite Tabs, 150 mg pinokinase, Aidan, Tempe, AZ, USA) in long-haul flights (7-8 hours), in high-risk subjects. A group of 300 subjects was included; 76 were excluded for several problems including concomitant treatments; 204 were randomized into 2 groups (active treatment or placebo) to evaluate the effects of prophylaxis with Flite Tabs. An exercise program was used in both groups. The femoral, popliteal, tibial, and superficial veins were scanned with ultrasound before and within 90 minutes after flights. Of the included subjects, 92 of 103 controls and 94 of 101 treated subjects completed the study. Dropouts were due to connection problems. Age, gender, and risk distribution were comparable in the groups. In the treatment group, no DVT was observed. In the control group, 5 subjects (5.4%) had a DVT and there were 2 superficial thromboses (7 events in 92 subjects; 7.6%). At inclusion, edema was comparable in the 2 groups. After flights there was an increase in score in controls (+12%) in comparison with a decrease (-15%) in the Flite Tabs group (the difference in variation was statistically significant). Intention-to-treat analysis for thrombotic events shows 18 failures in controls (11 lost to follow-up + 7 thrombotic events) of 92 subjects (19.6%) in comparison with 7 failures (of 94 subjects, equivalent to 7.4%) in the treatment group (p < 0.05). Events were asymptomatic. In conclusion, Flite Tabs were effective in reducing thrombotic events and in controlling edema in high-risk subjects in long flights. PMID- 14565629 TI - A new noninvasive method for the accurate and precise assessment of varicose vein diameters. AB - The feasibility and reproducibility of a new ultrasonic method for the direct assessment of maximal varicose vein diameter (VVD) were evaluated. A study was also performed to demonstrate the capacity of the method to detect changes in venous diameter induced by a pharmacologic treatment. Patients with varicose vein disease were recruited. A method that allows the precise positioning of patient and transducer and performance of scans in a gel-bath was developed. Maximal VVD was recorded both in the standing and supine positions. The intraassay reproducibility was determined by replicate scans made within 15 minutes in both positions. The interobserver variability was assessed by comparing VVDs measured during the first phase baseline examination with those obtained during baseline examinations in the second phase of the study. The error in reproducibility of VVD determinations was 5.3% when diameters were evaluated in the standing position and 6.4% when assessed in the supine position. The intramethod agreement was high, with a bias between readings of 0.06 +/- 0.18 mm and of -0.02 +/- 0.19 mm, respectively, in standing and supine positions. Correlation coefficients were better than 0.99 in both positions. The method appears to be sensitive enough to detect small changes in VVDs induced by treatments. The proposed technique provides a tool of potential valid use in the detection and in vivo monitoring of VVD changes in patients with varicose vein disease. The method offers an innovative approach to obtain a quantitative assessment of varicose vein progression and of treatment effects, thus providing a basis for epidemiologic surveys. PMID- 14565630 TI - Clustered features of the metabolic syndrome and the risk for increased aortic pulse wave velocity in middle-aged Japanese men. AB - The association between different features of the metabolic syndrome (MS) (obesity, hypertension, hypercholesterolemia, low high-density lipoprotein cholesterol level, hypertriglyceridemia, high fasting plasma glucose level, and hyperuricemia) and the risk for increased aortic pulse wave velocity (PWV) of > or = 8.0 m/sec was examined in 2431 Japanese men aged 35 to 54 years who were not taking antihypertensive medication. After controlling for age, cigarette smoking, and alcohol intake, the odds ratios for increased aortic PWV in subjects with 1, 2, 3, and > or = 4 features of the MS, compared with those without features of the MS, were 1.35 (95% CI, 0.86 to 2.11), 1.90 (95% CI, 1.18 to 3.06), 1.57 (95% CI, 0.89 to 2.76), and 2.38 (95% CI, 1.26 to 4.49), respectively (p for trend = 0.003). A 9-year longitudinal study was also performed to prospectively examine the association between clustered features of the MS and the development of increased aortic PWV in 2073 men without aortic stiffness with a PWV < 8.0 m/sec and without antihypertensive medication during the follow-up period. The multivariate-adjusted hazard ratios for the incidence of increased aortic PWV in subjects with 1, 2, 3, and > or = 4 features of the MS, compared with those without features of the MS, were 1.39 (95% CI, 1.10 to 1.77), 1.46 (95% CI, 1.1 1 to 1.92), 1.75 (95% CI, 1.27 to 2.40), and 2.22 (95% CI, 1.52 to 3.25), respectively (p for trend < 0.001). These results suggest that clustered features of the MS are closely associated with the risk for increased aortic PWV in middle aged Japanese men. PMID- 14565631 TI - Mitral annular calcification and aortic valve calcification may help in predicting significant coronary artery disease. AB - Mitral annular calcification (MAC) and aortic valve calcification (AVC) are manifestations of atherosclerosis. To determine whether mitral annular calcification and aortic valve calcification detected by transthoracic echocardiography (TTE) might help in predicting significant coronary artery disease (CAD), 123 patients with significant CAD and 93 patients without CAD detected by coronary angiography were investigated. MAC and AVC identified CAD with a sensitivity and specificity of 60.2%, 55.9% and 74.8%, 52.7%, respectively, and with a negative and a positive predictive values of 51.5%, 64.3% and 61.3% and 67.6%, respectively. The positive predictive value of MAC was greater than gender, hypertension, and hypercholesterolemia. AVC showed a positive predictive value greater than gender, hypertension, family history, and hypercholesterolemia. The negative predictive values of MAC and AVC for CAD were greater than those of all risk factors except diabetes mellitus. In conclusion, presence of MAC and AVC on TTE may help in predicting CAD and should be added to conventional risk factors. Absence of MVC and AVC is a stronger predictor for absence of CAD than all conventional risk factors, except diabetes mellitus. Patients with MAC and AVC should be taken into consideration for the presence of significant CAD and thereby for diagnostic and therapeutic interventions in order to improve the prognosis. PMID- 14565633 TI - In-hospital complications of percutaneous intraaortic balloon counterpulsation. AB - Complications related to intraaortic balloon counterpulsation pumping (IABP) remain a problem despite the development of small caliber balloon catheter shafts and introducer sheaths. The authors report their experience in counterpulsation related complications of 201 consecutive patients who underwent 212 percutaneous counterpulsation balloon insertions from June 1989 to June 1996 by use of balloons with 8-9.5 French shafts. Of these, 82% were men and 36 (18%) were women, with a mean age of 61 +/-12 years. Indications for counterpulsation were acute myocardial infarction (AMI) (67%), severe left ventricular failure without AMI (20%), dilated cardiomyopathy (4%), unstable angina (3%), high-risk supported percutaneous coronary angioplasty (2%), and others (4%). IABP was instituted at the bedside in the intensive care unit in 82 patients (39%) and in the catheterization laboratory in 130 (61%). Median duration of counterpulsation was 48 hours (range 30 minutes to 25 days) with successful weaning from counterpulsation in 70% (148 of 212) of procedures. Overall in-hospital mortality rate was 45% (90 of 201). The overall complication rate was 22/212 (10.4%). Major complications were present in 10/212 procedures (4.7%): 6 patients with limb ischemia (1 death directly attributed to this complication, 1 with associated septicemia and limb amputation, 3 requiring surgical thromboembolectomy, and 1 with persistent limb ischemia treated medically until his death caused by intractable left ventricular failure), 2 with important bleeding (1 fatal despite vascular surgical repair and 1 requiring blood transfusion) and 2 with balloon rupture requiring vascular surgery. Minor complications were present in 12 procedures (5.7%), 6 with limb ischemia, 3 with local bleeding, and 3 with catheter dysfunction. All of these resolved after balloon removal and required no further intervention. When limb ischemia did develop it occurred after a median delay of 24 hours following balloon insertion (range 2 to 98 hours). The only predictor of limb ischemia among baseline clinical and procedure-related variables was an age greater than 60 years. Compared with previous recent studies, the rate of complications observed in this study performed with small balloon catheters was acceptably low. Limb ischemia was the most frequent complication, often occurred early, and required further intervention in half the cases. PMID- 14565632 TI - Subclinical hypothyroidism and lipid abnormalities in older women attending a vascular disease prevention clinic: effect of thyroid replacement therapy. AB - The authors evaluated the frequency and type of lipid disorders associated with subclinical hypothyroidism (SH) in older women referred to their university vascular disease prevention clinic. They also assessed the results of thyroid replacement therapy. Fasting serum lipid profiles and thyroid function tests were measured in 333 apparently healthy women (mean age: 71.8 +/- 7 years). These women were divided into 3 groups: group I: 60-69 years old (n = 132); group II: 70-79 years old (n = 153); group III: 80-89 years old (n = 48). SH was defined as a serum thyrotropin concentration higher than 3.20 mlU/mL with a normal free thyroxine concentration. The prevalence of SH was 7.5%. Thyrotropin was higher than 3.20 mU/mL in 25 women; 7 (5.3%), 14 (9.2%), and 4 (8.3%) in groups I, II, and III, respectively. Low-density lipoprotein cholesterol (LDL-C) concentrations were higher in the women with SH (p = 0.037). The mean values of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), TC/HDL-C ratio, lipoprotein (a) (Lp[a]), apolipoprotein A-I (apo AI) apolipoprotein B100 (apo B) and apo B/apo A ratio were higher and triglycerides (TG) were lower, compared with those with normal levels of thyrotropin. However, none of these differences reached significance. Restoration of euthyroid status (thyroxine: 50-100 microg/day) in 17 SH women significantly improved TC (p = 0.017), LDL-C (p = 0.014), TC/HDL-C (p = 0.05), LDL-C/HDL-C (p = 0.03), apo B (p = 0.013), and Lp(a) (p = 0.0005) values. SH is relatively common in older women attending a vascular disease prevention clinic. Thyroid hormone replacement therapy significantly improved serum lipids. In particular, the reduction in LDL-C and Lp(a) concentrations may be of clinical benefit. PMID- 14565634 TI - Clinical logistics in 24-hour ambulatory electrocardiographic monitoring. AB - In total, 493 ambulatory ECG recordings were studied. Women were preponderant (62.3% vs 37.7%). The average age of women and men patients was 66.9 and 64.7 years, respectively. Of the ECGs studied, 71.4% showed abnormalities and 28.6% appeared completely normal. Urgent abnormalities were noted in 1.4% of the recordings and significant abnormalities were present in 14.6%. Subjective complaints were noted in their logbooks by 18.8% of patients, but correlation of complaints with the electrocardiographic abnormalities was noted in only 1.2% of cases. The attending cardiologist concluded that 23.9% of the tests supported reasons of valid necessity for performance. Two hundred seventy-three recordings were classified as electrocardiographically abnormal (55.4%) but were clinically insignificant. General practitioners requested 59.8% of the tests versus 40.2% by specialists. Preponderant abnormalities included premature atrial and ventricular contractions, supraventricular tachycardia, and atrial fibrillation. Less frequent abnormalities included ventricular tachycardia (4.6%), atrial flutter, atrioventricular block, artificial pacemaker rhythm, nodal rhythm, and intermittent bundle branch block. PMID- 14565635 TI - Increased systemic and regional coagulation activity in patients with mitral stenosis and sinus rhythm. AB - A hypercoagulable state has been reported in patients with mitral stenosis (MS) and sinus rhythm (SR). However it has been suggested that the coagulation activity may be increased only within the left atrium in MS, with normal peripheral blood levels. The aim of the present study was to assess regional left atrial and systemic coagulation activities by measuring PF1+2 in patients with severe mitral stenosis and sinus rhythm, normal blood clotting times, and no left atrial thrombus. The study was conducted in 25 consecutive patients with moderate to-severe MS and sinus rhythm who underwent percutaneous balloon mitral valvuloplasty. Transesophageal echocardiography was performed before the valvuloplasty procedure in all patients to exclude the presence of left atrial thrombus and left atrial spontaneous echo contrast (LASEC). There were no statistically significant differences between LASEC-positive and LASEC-negative patients with respect to age, gender, fibrinogen levels, prothrombin time, mitral valve area, mean mitral gradient, pulmonary artery pressure (in all p > 0.05). Regional (left atrial) PF1+2 levels of both LASEC-positive and LASEC-negative patients were significantly elevated when compared to control subjects (p < 0.01). Statistically significant elevated systemic level of PF1+2 was observed only in LASEC-positive patients when compared to control subjects (p < 0.01, p > 0.05, respectively). In conclusion patients with severe mitral stenosis and SR have increased regional coagulation activity in both LASEC-negative and LASEC positive groups. Although this increased regional coagulation activity has been reflected in peripheral blood of LASEC-positive patients, it has not been reflected in peripheral blood of LASEC-negative patients. PMID- 14565636 TI - Clinical perspectives of primary cardiac lymphoma. AB - Primary cardiac lymphoma is a very rare malignancy, which is typically of a non Hodgkin type, and involves only the heart and pericardium with no or minimal evidence of extracardiac involvement. Primary cardiac lymphoma account for about 1% of the primary cardiac tumors and 0.5% of the extranodal lymphomas. On the other hand, disseminated lymphoma with cardiac involvement can occur in up to 20% of patients with lymphoma. About 80% of cases of the primary cardiac lymphoma in immunocompetent hosts are of diffuse B-cell lymphoma, and in patients with immunodeficiency states, small noncleaved or immunoblastic lymphomas are more frequent. The right atrium and right ventricle are the 2 most frequently involved sites. Clinical presentation is heterogeneous and is generally related to the site of involvement in the heart. The diagnosis is suspected when patients present with a cardiac mass or an unexplained refractory pericardial effusion. A thorough workup should include transthoracic and transesophageal echocardiography, computed tomography, and magnetic resonance imaging. Diagnosis is confirmed by cytology of the serous fluid from pericardial or pleural effusion or biopsy of the pericardial mass or endomyocardial tissue. The exploratory thoracotomy should not be delayed if indicated. Chemotherapy has been used alone or combined with radiotherapy. Similarly, palliative cardiac surgery has been performed, mainly for tumor debulking. Combination of chemotherapy and radiation therapy is considered as the treatment of choice. The survival is generally less than a month without treatment but has been prolonged up to 5 years with palliative treatments in selected cases. PMID- 14565637 TI - Protein S deficiency presenting as deep vein thrombosis--a case report. AB - A rare case of spontaneous bilateral internal jugular vein (IJV) thrombosis is presented in absence of oral infection, malignancy, or intervention. Swelling of face, neck, and both upper limbs developed in a 30-year-old man 1 month before admission and after 4 weeks of use of nonpadded crutches. Computed tomography of the neck showed thrombus occluding both internal jugular veins extending to the opening of the superior vena cava. Protein S activity was found to be 30.3%, less than half of the lower limit of normal. Fibrinogen was 450 mg%, higher than normal. Protein C and antithrombin III were normal. The patient was treated conservatively, and discharged on aspirin 150 mg/day. Follow-up after 3 months showed no complications. It was concluded that the outcome in IJV thrombosis, caused by protein S deficiency, is usually good. The advantage of being aware of diagnosis is that physician can be more vigilant for potential complications and treat them earlier. PMID- 14565638 TI - Restoration of endothelial function after repaired coarctation of the aorta--a case report. AB - According to current literature, there is no clinical report regarding coronary spasm in the presence of aortic coarctation. There are no cases in which endothelial function was restored after the repair of aortic coarctation. A 29 year-old woman with aortic coarctation and acute myocardial infarction due to coronary spasm experienced restoration of endothelial function after the repair of aortic coarctation. This case may increase knowledge of the mechanism of coronary damage in human coronary arteries not only with a coarcted aorta but also with hypertensive heart. PMID- 14565639 TI - Chordae tendineae rupture resulting in pulmonary edema in a patient with discrete subvalvular aortic stenosis--a case report and literature review. AB - This report concerns an apparently healthy elderly woman who presented with gradually worsening mitral regurgitation secondary to chordae tendineae rupture leading to pulmonary edema in the presence of discrete subvalvular aortic stenosis with a severe gradient reflecting the left ventricular outflow tract obstruction. The gradual worsening of heart failure took place parallel to the increase in severity of mitral regurgitation in a short period. The patient underwent successful mitral valve replacement with myectomy. Surgical inspection revealed rupture of the chordae tendineae to the posterior leaflets without any significant primary intrinsic disease of the mitral valve. The predominant mechanism of chordae tendineae rupture in this patient with discrete subvalvular aortic stenosis is a severe pressure gradient. It is suggested that increased awareness of chordae tendineae rupture as a cause of mitral regurgitation and the prompt use of appropriate diagnostic tools may facilitate the timely recognition of this potentially fatal, but treatable, cause of mitral regurgitation in patients with left ventricular outflow tract obstruction. PMID- 14565640 TI - Contrast transesophageal echocardiography in diagnosing congenital enlargement of the right atrium--a case report. AB - Congenital malformation of the right atrium or the coronary sinus is rare, and cases are classified into 1 of the following 4 categories: (1) congenital enlargement of right atrium, (2) single diverticulum, (3) multiple diverticula of the right atrium, and (4) diverticulum of the coronary sinus. This report presents a 63-year-old man with cardiomegaly and no chest symptoms. A chest radiograph revealed an enlarged cardiac silhouette with a prominent right heart border. Although a transesophageal echocardiography revealed marked enlargement of the right atrium, neither further anomaly nor massive regurgitation was observed. The systolic pulmonary artery pressure derived from the peak velocity of mild tricuspid regurgitation was 38 mm Hg. Secondary enlargement of the right atrium due to atrial septal defect or pulmonary venous connection anomaly was deemed negligible by use of transesophageal contrast echocardiography, and primary enlargement of the right atrium was confirmed. Transesophageal echocardiography using ultrasound contrast was determined to be feasible for diagnosing congenital malformation of the right atrium. PMID- 14565641 TI - Congestive heart failure due to traumatic arteriovenous fistula--two case reports. AB - Arteriovenous fistulas are abnormal connections between the high-pressure and high-resistance arterial system and the venous system with opposite features. Due to its lower resistance, the blood preferentially flows via the fistula rather than through the capillary bed. The amount of shunt flow depends on its size and proximity to the heart. Due to the increase in circulating volume, progressive dilation develops in the whole vascular system proximal to the shunt. Cardiomegaly and venous distention may return to normal after surgical repair of this vascular abnormality. Two cases are presented of heart failure due to traumatic arteriovenous fistula, one of which was between the right renal artery and inferior vena cava and the other between the left renal artery and renal vein. PMID- 14565642 TI - Pneumomediastinum and cervical emphysema associated with unusual clinical and electrocardiographic manifestations--a case report. AB - This is the first case reported of combined cervical emphysema and pneumomediastinum associated with unusual electrocardiographic and local neurologic findings. These may be the result of an increase in intracervical and intrathoracic pressure induced by the dissecting air. PMID- 14565643 TI - Flow cytometric detection of minimal residual disease in acute lymphoblastic leukemia. AB - Assessment of minimal residual disease (MRD) during the first months of therapy gives information on the timely response to treatment, and proves to be a powerful and independent indicator of treatment outcome in patients with acute lymphoblastic leukemia (ALL). Immunological evaluation by flow cytometry (FCM) is one of the most attractive approaches to this. The present review summarizes the historical development of this approach over the last 20 years, and shows that current methodology is based on the existence of leukemia-associated patterns of derangement in antigen expression with respect to normal differentiation or location of occurrence. Recent clinical studies are summarized which proved that FCM is applicable to more than 90% of patients with ALL and gives prognostic information comparable to polymerase chain-reaction (PCR)-based technology. Ongoing efforts based on parallel application of both technologies are explained which are designed to clarify which approach bears the best cost-relevance ratio in order to be broadly used in the future for risk assessment and tailoring of treatment modalities. Concluding perspectives relate to further technical developments like usage of peripheral blood (PB) instead of bone marrow (BM), absolute quantification, or strategic placement of investigative time-points, which may allow to simplify the MRD approach and thus augment it's economic efficiency. PMID- 14565644 TI - Cytokine-induced killer cells: NK-like T cells with cytotolytic specificity against leukemia. AB - Cytokine-induced killer (CIK) cells are a unique population of cytotoxic T lymphocytes (CTL) with the characteristic CD3+CD56+ phenotype. These cells have demonstrated higher proliferative and cytolytic activities in comparison to the reported CD3-CD56+ lymphokine activated killer (LAK) cells that are essentially activated natural killer (NK) cells. CIK cells are non-MHC-restricted in target cell recognition and killing. We have shown the feasibility of generating CIK cells from a series of marrow samples of patients with acute myeloid leukemia (AML) collected at diagnosis. At maturity, the CIK cells exhibit potent cytotoxicity against autologous AML targets as well as allogeneic myeloid leukemia cells, regardless of the HLA types of these targets. This observed cytotoxicity is not entirely due to NK cells as prior pre-absorption of the NK cells cytolytic activities does not abolish the subsequent cytotolytic activities against leukemic targets. It has also been reported by others that CIK cells are cytolytic against chronic myeloid leukemia (CML) cells, both in vitro and in the SCID mouse tumor model. In a mouse transplant model across MHC barrier, the CIK cells generated from the donor do not induce graft vs. host disease as observed for unfractionated donor splenocytes. In comparison to untreated control mice, the infusion of CIK cells results in the prolonged survival of murine leukemia bearing mice. CIK cells also express CD94, part of the NK receptor comprising of CD94-NKG2 heterodimer. However, only low level of the killer immunoglobulin-like receptors are expressed by the CIK cells. In addition, as reported for the classical CTL, CIK cells could interact with dendritic cells (DC) to result in the enhancement of cytotolytic activities against tumor cells. The characteristic biological properties of the CIK cells would, therefore, enable them to be exploited for anti-leukemic therapy. PMID- 14565645 TI - New insights in myeloma-induced osteolysis. AB - Multiple myeloma (MM) is a plasma cell malignancy localized in the bone marrow (BM) and characterized by a high capacity for bone destruction. Almost all patients with MM have early osteolytic lesions, which result mainly from increased bone resorption related to stimulation of osteoclast recruitment and activity in the immediate vicinity of myeloma cells. The recent discovery of Osteoprotegerin (OPG) and the subsequent identification of its ligand RANKL have provided new insights in the regulation of osteoclastogenesis. The ratio OPG/RANKL is critical for the regulation of bone remodeling maintaining the balance between osteoblastic and osteoclastic activity. This review summarizes the new concept that myeloma cells induce in bone environment an imbalance in the OPG/RANKL system responsible for osteolysis observed in patients. Indeed, myeloma cells increase in bone environment the expression of the potent osteoclastogenic factor RANKL and decrease the osteoprotective factor OPG production. Biological mechanisms involved in these processes are discussed. Furthermore, the chemokines MIP-1alpha and MIP-1beta belonging to the RANTES family are potent osteoclastogenic factors produced by myeloma cells and participate in myeloma associated bone disease. These data open new avenues for the treatment of bone disease in MM and highlight the promising therapeutical interest of RANKL inhibitors (OPG and RANK-Fc) and MIP-1 inhibitors in the management of myeloma associated osteolysis, besides bisphosphonates. PMID- 14565646 TI - Hemopoietic precursors and development of dendritic cell populations. AB - The antigen presenting dendritic cells (DCs) are bone marrow (BM) derived cells. Despite their common functions of antigen-processing and T-lymphocyte activation, DCs are diverse in surface markers, migratory patterns and cytokine output. These differences can determine the fate of the T cells they activate. Several subsets of mature DCs have been described in both mouse and human, but tracing the origin of these specialised DC subsets has not been a trivial task. The original concept that all DCs were of myeloid origin was questioned by several recent studies, which demonstrated that in addition to the DCs derived from conventional myeloid precursors, some DCs could also be efficiently generated from lymphoid-restricted precursors. Moreover, it has been shown that both myeloid-restricted and lymphoid restricted precursors were able to generate DC subsets with similar surface phenotype. These observations demonstrate the existence of both myeloid- and lymphoid-derived DC lineages and suggest an early developmental flexibility of DC precursors. The downstream points where the DC sub-lineages branch off from the conventional myeloid and lymphoid precursors, and the cytokines and environmental factors required for inducing their specialised functions are yet to be determined. PMID- 14565648 TI - Treatment of polycythemia vera and essential thrombocythemia: the role of pipobroman. AB - Pipobroman (PB) is a neutral amide of piperazine with a chemical structure close to that of alkylating agents, although the exact mechanism of action of PB has not been demonstrated. PB has well documented clinical activity in polycythemia vera (PV) and essential thrombocythemia (ET). Recent long-term follow-up studies on PV and ET patients receiving PB have facilitated the definition of the risk of late transformation into myelofibrosis with myeloid metaplasia (MMM) or acute leukemia (AL). This report gives an overview of the treatment with PB in patients with PV and ET focusing on clinical activity, administration dose and schedule, toxicity, impact on short- and long-term complications. From our experience and from the data reported in the literature the high clinical activity of PB in both PVand ET becomes evident. This drug allows, within 3 months, to attain a response in more than 90% of patients, without clinically relevant toxicities. The 10 years risk of thrombosis of patients treated with PB is about 15%, similar to that registered with hydroxyurea, the most widely used agent in PVand ET. The antiproliferative activity of PB on bone marrow megakaryocytes seems of particular value in lowering the occurrence of post-PV and post-ET MMM, whose risk (< 4% at 10 years) is the lowest registered with available treatments. The 10-year risk of acute leukemia with PB is 5% in PVand 3% in ET, which is only slightly higher than that expected as a natural evolution of the disease. In conclusion, the use of PB is a definite alternative to hydroxyurea in patients with PV and ET at high risk of thrombosis. PMID- 14565649 TI - Thalidomide for the treatment of acute myeloid leukemia. AB - In analogy to solid neoplasms, accumulating data suggest the requirement of angiogenesis also for the development and progression of hematopoietic malignancies including acute myeloid leukemia (AML). Inhibition of increased microvessel density in bone marrow (BM) might be a promising target for pharmacological interventions aimed at reducing disease activity. Among the putative inhibitors of angiogenesis, thalidomide has demonstrated a considerable efficacy in myelodysplastic syndromes (MDS) and AML with overall response rates up to 56% and 25%, respectively. Responders experienced hematologic improvements with increased hemoglobin and platelet counts resulting in temporary transfusion independence. In AML, partial responses--defined as reduction of the leukemic blast cell infiltration of at least 50% in BM--occurred in four of 20 patients after one month of thalidomide administration in a previous phase I/II study. Additionally, we observed a long-term response in one AML patient of more than 20 months, meanwhile fulfilling the criteria of complete remission. The decrease in leukemic blast infiltration in BM of responders was accompanied by a significant reduction of the microvessel density. Overall adverse events caused by the drug consisted mainly of fatigue, constipation, skin rash and polyneuropathy with a tolerable dose of 200-400 mg p.o. per day. In conclusion, thalidomide as a single agent has significant anti-leukemic activity with some evidence for anti angiogenic effects in BM, although the precise mechanism of action remains to be elucidated. PMID- 14565647 TI - Interleukin-6, CD45 and the src-kinases in myeloma cell proliferation. AB - Multiple myeloma (MM) is a proliferative disorder of monoclonal plasma cells which accumulate in human bone marrow, and myeloma cells proliferate in response to a cytokine, interleukin-6 (IL-6). We recently found that MPC-1- CD49e- immature myeloma cells expressing CD45 form a proliferating population in MM. IL 6 activates at least two intracellular pathways including signal transducer and activator of transcription 3 (STAT3) and extracellular signal-regulated kinase 1/2 (ERK1/2) following the activation of Janus kinases (JAKs) via its receptor complexes composed of the IL-6 receptor alpha chain and gp130. Although the roles of CD45 have been extensively studied for antigen receptors in B and T cells, its physiological consequences in other hematopoietic cells remain largely unknown. Myeloma cells expressing CD45 antigens which contain the activation of src family protein-tyrosine kinases (PTKs) independent of IL-6 stimulation proliferate in response to IL-6, whereas the proliferation of CD45- cells which lack a considerable activity of the src family PTKs is not promoted by IL-6. The STAT3 and ERK1/2 pathways are similarly activated by IL-6 in both cells either expressing or not expressing CD45. In this review, we argue a novel mechanism of proliferation of myeloma cells, in that the activation of both STAT3 and ERK1/2 is not sufficient for IL-6-induced proliferation which further requires IL-6 independent activation of the src family kinases associated with CD45 phosphatase. We propose that the cellular context, such as CD45 expression and src family kinase activation, is crucial for myeloma cells to proliferate in response to IL-6. PMID- 14565650 TI - Central venous catheter-related complications in patients with hematological malignancies: a retrospective analysis of risk factors and prophylactic measures. AB - We retrospectively analyzed the incidence of thrombotic and infectious complications in relation with the use of central venous catheters (CVCs), in a series of patients with hematological malignancies and low platelet and leucocyte counts. PATIENTS AND METHODS: 126 patients with hematological malignancies were analyzed. A total of 207 CVCs were implanted: 137 centrally (CICCs) and 70 peripherally (PICCs). The median duration of the CVCs was 19 days for a total of 4051 catheter-days. Antithrombotic prophylaxis was unfractionated heparin (UFH), 2,500 IU daily by 24 h continuous infusion in 169 CVCs, low molecular weight heparin (LMWH), 3,800 IU daily by single bolus intravenous injection (i.v.) in 21 and warfarin in one. No prophylaxis was given in 16 CVCs. Thrombotic complications developed in 15.5% of the CVCs (7.9 events/1000 catheter days), and the frequency of infectious complications was 10.6% (5.2 events/1000 catheter days). On multivariate analysis thromboses were more frequent and earlier with PICCs than CICCs (p = 0.0001), and in patients on UFH (16.6%) than in LMWH prophylaxis (4.7%), but the last difference was not statistically significant. In conclusions the incidence of thrombotic complications in our series was comparable to that observed in non-thrombocytopenic patients and was significantly higher in those carrying PICC than CICC (p = 0.0001). There were fewer thrombotic events in the patients receiving i.v. LMWH prophylaxis than in those receiving i.v. UFH. The use of anticoagulants was safe and not associated with hemorrhages. PMID- 14565651 TI - Open-label, randomized study of pegfilgrastim vs. daily filgrastim as an adjunct to chemotherapy in elderly patients with non-Hodgkin's lymphoma. AB - Pegfilgrastim is composed of the protein filgrastim to which a 20-kDa polyethylene glycol (PEG) is covalently bound at the N-terminal residue resulting in decreased renal clearance and increased plasma half-life compared with filgrastim. This open-label, randomized, phase 2 study compared two doses of single administration pegfilgrastim (60 and 100 microg/kg) with daily doses of filgrastim (5 microg/kg/day) or no cytokine treatment after standard CHOP (cyclophosphamide, doxorubicin, vincristine and prednisolone) chemotherapy for non-Hodgkin's lymphoma in 50 elderly patients. The primary endpoint was the duration of grade 4 (severe) neutropenia (absolute neutrophil count < 0.5 x 10(9)/l) in cycle 1. Duration of grade 4 neutropenia in cycle 1 was 2.2 (SD 1.2), 1.5 (SD 1.1), 0.8 (1.2) and 5.0 (2.0) days for patients who received pegfilgrastim 60 microg/kg, pegfilgrastim 100 microg/kg, filgrastim 5 microg/kg and no cytokine, respectively. The baseline characteristics of the pegfilgrastim and filgrastim groups were imbalanced with increased bone-marrow involvement and prior therapy in the former. When the treatment groups were balanced for these risk factors, duration of grade 4 neutropenia was comparable with 2.0 and 3.0 vs. 0.6 and 0.5 days for pegfilgrastim 100 microg/kg and filgrastim patients with and without these risk factors, respectively. The incidence of febrile neutropenia (defined as ANC < 0.5 x 10(9)/l and temperature > 38.2degrees C) was low (10% of patients). Pegfilgrastim was well tolerated with a safety profile similar to daily filgrastim. Once per chemotherapy cycle administration of pegfilgrastim was comparable to filgrastim in this clinical setting. PMID- 14565652 TI - CD34 positive selection as prophylaxis against graft versus host disease in allogeneic peripheral blood stem cell transplantation. AB - We report our experience of CD34 positive selection as a means of graft T-cell depletion (TCD) in 14 consecutive HLA-identical Peripheral blood stem cells (PBSC) allografts as prophylaxis against graft versus host disease (GVHDp). CD34 positive selection was performed by immunomagnetic separation achieving a median CD34 and T-cell dose of 4.17 (range 1.4-8.50) x 10(6)/kg and 1.89 (range 0.92 13.18) x 10(4)/kg, respectively, in the graft. This represents 4-log depletion of T-cells. The median time to achieve a neutrophil count of 0.5 x 10(9)/l was 15 days and to achieve a platelet count of 50 x 10(9)/l was 20 days. Only four patients developed acute GVHD at a median of 41 days but this was exclusively mild grade I cutaneous disease and settled with oral steroids. Four patients, all of whom had AML, relapsed or progressed after transplant at a median of 161 (range 109-311) days. One of these had been transplanted in early relapse (9% blasts) whilst another was in second remission. The remaining 10 patients are alive and well. The median progression free survival for the whole population is 69% at 686 days. We conclude that CD34 positive selection by immunomagnetic separation in HLA-identical PBSC allografting achieves significant TCD with clinically trivial acute GVHD, prompt engraftment and an acceptable disease relapse risk. PMID- 14565653 TI - A randomized trial of induction therapy (daunorubicin, vincristine, prednisone versus daunorubicin, vincristine, prednisone, cytarabine and 6-thioguanine) in adult acute lymphoblastic leukemia with long-term follow-up: an Eastern Cooperative Oncology Group Study (E3486). AB - In this study of previously untreated adult acute lymphocytic leukemia (ALL) performed by the Eastern Cooperative Oncology Group, patients were randomized to induction therapy with either DVP (daunorubicin 45 mg/m2 daily, days 1, 2 and 3; prednisone 60 mg/m2 daily orally days 1-35; and vincristine 2 mg intravenously on days 1, 8, 15 and 22) or DATVP (daunorubicin 60 mg/m2 daily, days 1, 2 and 3; cytarabine 25 mg/m2 intravenous bolus followed by 200 mg/m2 daily as a continuous infusion on days 1-5; 6-thioguanine 100 mg/m2 orally every 12 h on days 1-5; vincristine 2 mg intravenously on days 1 and 8; and prednisone 60 mg/m2/day orally, days 1-7. Complete responders to both regimens received the same post remission therapy, which consisted of a single course of cytarabine 3 gm/m2 infused over 1 h every 12 h for 12 doses. One month later those patients still in remission received six cycles of consolidation therapy with MACHO (cyclophosphamide 650 mg/m2, doxorubicin 40 mg/m2 vincristine 2mg all intravenously on day 1 with prednisone 100 mg/m2 orally daily on days 1-5. Methotrexate 200 mg/m2 intravenously and L-asparaginase 6000 U/m2 were given on day 22 and each course was given every 5 weeks. A single dose of intrathecal methotrexate was also given with each MACHO course. There were 276 evaluable patients randomized in this study. Complete response rates were 71% for DVP and 58% for DATVP. Median durations of complete response were 5.5 and 6.8 months, respectively. Median survival of all randomized patients was 14.4 months in each group. DATVP was more toxic than DVP. Intensification of treatment for adults with ALL may not improve outcome. Progress in the treatment of adults with ALL will require the identification of new agents for this neoplasm. PMID- 14565654 TI - Treatment of early clinically staged Hodgkin's disease with a combination of ABVD chemotherapy plus limited field radiotherapy. AB - The current management of early stage Hodgkin's disease (HD) is usually based on clinical staging, combined modality therapy and the use of less toxic chemotherapy regimens. This approach entails high cure rates, while ensures less long term toxicity with avoidance of laparotomy. The aim of this study was to assess the efficacy of a brief course of Adriamycin, Bleomycin, Vinblastine, Dacarbazine (ABVD) chemotherapy followed by limited field radiotherapy (RT) in favorable clinical stage (CS) I and IIA HD. Forty patients, aged 17-68 (median 34) years, with favorable CS I and IIA HD, without bulky mediastinal disease, have been treated with 4-6 (median 4) cycles of ABVD plus limited field RT. Twenty seven (67%) patients received 4 cycles of chemotherapy, while 13 received 5-6 cycles. Thirty five (87%) patients received limited field RT with dose 24-36 Gy and five (13%) received extended field with 36-46 Gy. All patients responded completely to chemotherapy. One patient experienced a relapse two months after the end of therapy. All patients are alive; 39 in continuous complete remission. With a median follow-up period of 44 months (range 18-101) the actuarial overall and progress free survival was 100 and 97% at 5 years. We did not observe any case of secondary leukemia or solid tumor. Pulmonary toxicity was mild in cases of mediastinal irradiation. Considering the short follow-up time and the small number of patients, the combination of a brief course of ABVD plus regional RT is a very efficacious treatment of favorable CS I and IIA HD with mild toxicity. However, long term survival data are needed, which could give confident answers regarding the risk of late therapy related complications, particularly second malignancies. PMID- 14565655 TI - Phase II trial of irinotecan (CPT-11) in relapsed or refractory non-Hodgkin's lymphomas. AB - CPT11, a camptothecin analogue, is a specific DNA topoisomerase I inhibitor, with activity in tumor cell lines with MDR expression. CPT11 has a broad spectrum of activity in solid tumors (especially in colorectal, gastric and small cell lung cancers). Early reports have shown that CPT11 could be active in non-Hodgkin's lymphomas (NHL) with low-dose schedules. To further evaluate the efficacy and toxicity of CPT11 in patients with refractory or relapsed NHLs, we conducted a phase II trial with escalated doses. PATIENTS AND THERAPY: From 04/98 to 05/01, 28 patients with NHL were enrolled. PATIENTS CHARACTERISTICS: M/F 21/7; median age: 56 years (range 28-72); Ann Arbor stage at the time of the study I/II and III/IV in 6 and 21 patients, respectively. Sixteen patients had refractory disease when they were enrolled in this phase II study and 8 patients were previously treated with high-dose therapy and stem-cell transplantation. CPT11 was administrated at the doses of 350 mg/m2 every 3 weeks. Six courses were given in patients who achieved CR, PR or stable disease. Patients were evaluated every 2 courses. If no grade II or more toxicity was observed after the first course, escalated dose (500 mg/m2) was then undertaken. RESULTS: 19/28 patients received more than 2 courses of CPT11 and were evaluated for response. Nine patients received one course of therapy because of either progressive disease (n = 6), toxicity (n = 2) or refusal (n = 1). Ten patients received escalated dose (500 mg/m2). Complete remission and partial was achieved in 2/19 patients, stable disease in 7/19, and progressive disease in 10/19 patients. Median duration of responses was short (3 months, range 1-8 months). Seventy-five courses were evaluated for toxicity according to the WHO criteria. Diarrhea grade 2 or 3 occurred in 9/75 courses; cholinergic syndrome grade 2 in 3/75 courses; nausea grade 3 in 7/75 courses. Hematological toxicity: leucopenia grade 3 or 4 in 21/75 courses; thrombocytopenia grade 3 in 8/75 courses; infectious episodes grade 2 or 3 in 7/75 courses. In 2/7 courses with escalated doses, grade I/IV neutropenia occurred withoutother major toxicity. CONCLUSION: CPT11 has low activity in heavily pretreated NHLs. Responses were of short duration. PMID- 14565656 TI - Consolidation radiotherapy following brief chemotherapy for localized diffuse large B-cell lymphoma: a prospective study. AB - Many physicians administer involved field radiation therapy (RT) following brief chemotherapy for localized aggressive non-Hodgkin's lymphoma. Involved field irradiation usually implies treatment to the involved nodal regions with and without the contiguous lymphatic region, however, there is no agreements about its definition. Here we assess the appropriateness of RT irrespective of lymph node regions (localized field) following chemotherapy for patients with early stage diffuse large B-cell lymphoma. The localized field encompassed all original gross tumor volumes before chemotherapy with at least a 2- to 3-cm margin irrespective of lymphatic regions. We also evaluated the suitable radiation dose on the basis of response to chemotherapy. Twenty five eligible patients were treated with 3 cycles of chemotherapy (CHOP) followed by RT. All 25 patients had disease confined to Waldeyer's ring and/or cervical lymph nodes. Twenty two patients in complete response following chemotherapy received 30 Gy, and the remaining 3 in partial response received 40 Gy. With a median follow up of 42 months, both event free and overall survival rates at 2 years were 96.0%. There were no in-field recurrences, however, two patients experienced relapses. One developed central nervous system involvement and subsequently died of his disease. The other had mediastinal and submental lymph node relapse at 32 months, and is alive after salvage chemotherapy. Our study demonstrated that it should be possible to reduce treatment volume to less than the conventional involved field, and to limit the dose of RT in the range of 30-40 Gy. PMID- 14565657 TI - Post-transplant Burkitt's leukemia or lymphoma. Study of five cases treated with specific intensive therapy (PETHEMA ALL-3/97 trial). AB - Burkitt's lymphoma (BL) and Burkitt-like acute lymphoblastic leukemia (ALL) are uncommon lymphoproliferative disorders after solid organ or stem cell transplantation. Although their prognosis is considered to be poor, there are scarce data on the clinical characteristics and the response to specific therapies. We report the main clinical characteristics and the results of a specific intensive chemotherapy in 5 adult patients with postransplant BL/ALL3 included in the PETHEMA ALL3/97 protocol. Two patients died in induction, another died in consolidation phase and the remaining 2 patients are in continuous complete remission 6 and 18 months from the diagnosis. PMID- 14565658 TI - Pamidronate reduces skeletal events but does not improve progression-free survival in early-stage untreated myeloma: results of a randomized trial. AB - Ninety patients with untreated, stage I-II A myeloma, were randomised to receive or not monthly infusions of pamidronate (PMD) for 1 year, without additional therapies. Follow-up ranged from 36 to 72 months (median 51 months). Three years after the start of the treatment, the disease had progressed in 25% of PMD treated patients and in 26.8% of controls (p n.s). Median time-to-progression was 16 and 17.4 months, respectively (p n.s). Among the 21 patients who required chemo-radiotherapy, skeletal events (osteolytic lesions, pathological fractures and/or hypercalcemia) developed in 9/11 (81.8%) controls and in 4/10 (40%) of treated patients (p < 0.01). "Prophylactic" administration of PMD may decrease the development of skeletal events, but does not reduce the rate and the time of disease progression in early-stage myeloma. PMID- 14565659 TI - The influence of imatinib mesylate (STI571) used alone or in combination with purine nucleoside analogues on the normal and chronic myelogenous leukaemia progenitor cells in vitro. AB - Imatinib mesylate (STI571, Glivec), a signal transduction inhibitor used as a single agent demonstrates significant activity in patients with chronic myelogenous leukaemia (CML). Nevertheless, the interaction between STI571 and other antileukaemic drugs such as hydroxyurea, interferon alpha or cytarabine have also been investigated in order to further improve its effectiveness. In this study we have tried to answer the question if the combination of STI571 with purine nucleoside analogues (PNAs)- cladribine (2-CdA) and fludarabine (F-ara-A) intensifies the antiproliferative effect on granulocyte-macrophage progenitor cells (CFU-GM) from patients with CML as well as from normal persons. Our studies were based on the method of semisolid CFU-GM cultures in vitro. We added STI571 or PNAs singly to the culture, each of the drugs at three concentrations, as well as in combinations of the concentrations used. We showed that STI571 (0.5, 1.0 and 2.0 microM) used alone inhibited the colony growth of CML CFU-GM, as compared to CFU-GM derived from normal donors (p = 0.03; p = 0.0004; p = 0.0001). We also observed that STI571 used together with 2-CdA (5,10 and 20 microM) or F-ara-A (0.2, 0.4 and 0.8 microM) at all the combinations significantly inhibited the colony growth of CML CFU-GM, as compared either to the control or to STI571 used alone (p < 0.05). In addition, the differences between CML and normal CFU-GM colony growth inhibition after the use of the combination of the highest concentrations of STI571 either with 2-CdA or F-ara-A were statistically significant (p = 0.03 and p = 0.01, respectively). In conclusion, STI571 used together with both the PNAs had an additive effect on CML CFU-GM cells. However, further experimental and clinical studies concerning the usefulness of these combinations in the treatment of CML patients seem warranted. PMID- 14565660 TI - Prediction of high affinity class I-restricted multiple myeloma idiotype peptide epitopes. AB - Idiotypic determinants are potential patient-specific tumor antigens in multiple myeloma (MM). In this study, we have determined the DNA sequence of the variable region of the tumor immunoglobulin (Ig) in 6 patients with MM. We then selected high affinity class I-restricted T-cell peptide epitopes in tumor Ig using two different internet-based epitope prediction programs. High affinity binding peptides were identified by at least one program in 4 out of 6 patients. Of these 35 peptides, only 3 scored high by both analyses. Given that all 6 patients had expanded T-cell clones with a cytotoxic (CD57+CD8+CD28-perforin+) phenotype, known to be associated with a longer survival and postulated to recognise tumor epitopes, this analysis indicates that such clones are unlikely to be exclusively directed towards tumor immuoglobulin, and suggests the need to expand the scope of the search for tumor epitopes with the ability to stimulate cytotoxic T cells in vivo. PMID- 14565661 TI - The anti-leukemic Bruton's tyrosine kinase inhibitor alpha-cyano-beta-hydroxy beta-methyl-N-(2,5-dibromophenyl) propenamide (LFM-A13) prevents fatal thromboembolism. AB - The leflunomide metabolite analog alpha-cyano-beta-hydroxy-beta-methyl-N-(2,5 dibromophenyl)-propenamide (LFM-A13) is a rationally-designed specific inhibitor of the TEC family protein tyrosine kinase, Bruton's tyrosine kinase (BTK) which plays an important role in platelet physiology by regulating the glycoprotein GPVI-FcRgamma-coupled collagen receptor signaling pathway. At low micromolar concentrations, LFM-A13 inhibited collagen-induced ultrastructural changes indicative of activation. LFM-A13 inhibited collagen (but not thrombin, TRAP-6, or ADP)-induced platelet aggregation in a concentration-dependent fashion with an IC50 value of 2.8 microM. LFM-A13 was not toxic to mice when administered systemically at dose levels ranging from 1 to 100 mg/kg. At nontoxic dose levels, LFM-A13 prolonged the tail bleeding times of mice and improved event-free survival in two mouse models of agonist-induced invariably fatal pulmonary thromboembolism. To our knowledge, LFM-A13 is the first anti-thrombotic agent which prevents platelet aggregation by inhibiting BTK. PMID- 14565662 TI - 13q deletion in chronic lymphocytic leukemia: characterization of E4.5, a novel chromosome condensation regulator-like guanine nucleotide exchange factor. AB - We report the characterization of a new gene (E4.5) that maps at chromosome band 13q14.3, a chromosomal area frequently deleted in chronic lymphocytic leukemia (CLL) and in other lymphoid malignancies. E4.5 gene encodes for a 4 kb mRNA expressed in various tissues and has an open reading frame of 531 amino acids. The predicted E4.5 protein shows strong homology with the human regulator of chromosome condensation (RCC1) protein, the principal GTP exchange factor for Ran protein. The E4.5 protein contains a BTB domain in its N-terminus, a protein protein interaction motif. Therefore, we propose that E4.5 is a new member of the RCC1-related guanine nucleotide exchange factor (GEF) family with potent interaction with other proteins and unknown function. Until now, no tumor suppressor genes have been mapped in the 13q14.3 minimal deleted region (MDR) in patients with CLL. It has been proposed that loss of the 13q14.3 MDR may contribute to lymphoid neoplasia by altering the expression/function of genes located on 13q14.3 outside the MDR. The E4.5 is one of these genes with a potential role in the pathogenesis of CLL. PMID- 14565663 TI - Abundant expression of spliced HDM2 in Hodgkin lymphoma cells does not interfere with p14(ARF) and p53 binding. AB - Recently, comparative genomic hybridization (CGH)- and fluorescence in situ hybridization (FISH)-analyses of native Hodgkin and Reed-Sternberg (H&RS) cells extracted from Hodgkin lymphoma (HL) revealed a recurrent amplification of the HDM2 locus on chromosome 12. HDM2 is known to target, inactivate and to degrade p53. Wild type (wt) p53 protein is detected in high levels in HL. Simultaneously, stabilized wt p53 and spliced hdm2 transcripts have been observed in different tumors. Therefore, we examined the expression and structure of HDM2 in HL cell lines and possible effects on components of the p53 pathway. DNA integrity and induction potential of p53 was verified by DNA sequencing and detection of potential effector proteins (p21(WAF/CIP), HDM2) using immunofluorescence, respectively. All HL cell lines show an overexpression of HDM2 protein. Furthermore, several different spliced hdm2 transcripts (mdm-sv) including five new variants lacking a functional p53 binding site were characterized. If expressed, corresponding proteins were shown to be not restricted to the nucleus. Co-localization of the potential binding partners HDM2/p14(ARF) and HDM2/p53 was found in HL cell lines. We suggest that HDM2-sv have no significant disturbing influence on the interaction of these proteins. PMID- 14565664 TI - Non-radiolabelled PCR consensus primers and automatic sequencing enable rapid identification of tumor-specific V(H) CDR3 in aggressive B-cell malignancies. AB - Even though the results of current therapy are improved for B-cell acute lymphoblastic leukemia (B-ALL) and Burkitt's lymphoma (BL), prognosis of relapsed mature B-ALL and BL still remain extremely poor. In this study, we investigated the possibility of applying the use of non-radiolabelled PCR consensus primers and automatic sequencing for the rapid identification of the tumor-specific V(H) CDR3 nucleotide sequence, in mature B-ALL and BL. RNA was extracted from four consecutive, unselected samples from BL cases and three consecutive, unselected samples from mature B-ALL cases. The feasibility of the identification of the tumor-specific V(H) CDR3 nucleotide sequence was then assessed by using non radiolabelled PCR consensus primers with automatic sequencing. The tumor-specific V(H) CDR3 nucleotide sequence was successfully identified for all seven patients (3 mature B-ALL and BL). The time required was substantially lower than that of the other methods previously published, despite the poor quality of some of the samples. The procedure showed rapidity, reliability and reproducibility. The characteristics of the methodology applied widen the possibility of developing anti-idiotypic therapeutic strategies, even in these B-cell malignancies. PMID- 14565666 TI - Eosinophilic variant of chronic myeloid leukemia with vascular complications. AB - Eosinophilic variant of CML (eoCML) is a unique disease with a poor prognosis. Like the hypereosinophilic syndrome (HES), eoCML has no clinically identifiable reason for an increased eosinophil count in the peripheral blood. In contrast to HES, eoCML patients carry a distinct chromosomal abnormality. The bcr/abl fusion gene (Philadelphia chromosome) is the genetic basis of this clonal disease. Recently, eoCML has been separated from HES. Patients with eoCML frequently suffer organ damage including the heart and lungs. This damage is related to the release of eosinophilic granules in the blood, which results in fibrosis of the endothelial lining. We report a case of a peripheral vasculitis complicated by gangrene of the fingers in a patient with eoCML. Despite an almost complete response to CML treatment with Gleevac, combined with prednisone, aspirin and coumadin the patient sustained irreversible damage to the vascular lining of the distal arteries of the upper extremities. PMID- 14565665 TI - Acute myelofibrosis in a patient with diffuse large cell non Hodgkin's lymphoma and renal cancer. AB - Relapse after anthracycline based combination chemotherapy is frequently seen in patients with aggressive non Hodgkin's Lymphomas (NHL), whereas complications such as secondary leukemia or solid tumor rarely occur. We report a patient with diffuse large cell (DLC) NHL and concurrent renal cancer, who developed acute myelofibrosis (AMF) later in the course of her disease. This 60-year-old female patient presented with pancytopenia and a right sided renal mass. Diagnostic work up revealed severe bone marrow infiltration by DLC NHL and renal cancer T1N0M0G2. Cytogenetic and molecular evaluation of bone marow cells showed three distinct clones, (a normal 46XX karyotype, a ringed chromosome 7 and a third clone with an enlarged chromosome 2 as well as several fragments). The patient underwent nephrectomy and eventually received 6 cycles of CHOP 14 chemotherapy. Anemia persisted followed by severe granulocytopenia and thrombocytopenia 6 weeks later. Repeated bone marrow biopsy showed absence of lymphoma and/or cancer metastasis, but massive myelofibrosis with an increased number of atypical megakaryocytes. Considering the short clinical course and the absence of hepatosplenomegaly AMF was diagnosed. The concurrence of three distinctneoplasms within a short period of time as well as the complex cytogenetic aberrations found in her bone marrow cells reflect a strong individual susceptibility to malignant disease in this patient. PMID- 14565667 TI - Thymic hyperplasia following successful treatment for nodular-sclerosing Hodgkin's disease. AB - A young female patient showed up with mediastinal bulky disease and lymph node swelling in her left supraclavicular region. Clinical and histological investigations proved nodular-sclerosing Hodgkin's lymphoma. The patient received combined modality treatment according to the protocols of the German Hodgkin's Disease Study Group and achieved complete remission. Six months later the chest X ray and thoracic CT-scan showed mediastinal tumor masses suggesting relapsed Hodgkin's disease. Surprisingly, the histological investigation showed thymic hyperplasia as well as the absence of any signs of Hodgkin lymphoma. Thymic hyperplasia is well known as a potential differential diagnosis of mediastinal space-occupying lesions and also as a long-term complication in patients cured of Hodgkin's disease. A detailed case report and a complete review of literature are provided. PMID- 14565669 TI - Clonal T-helper lymphocytes and elevated IL-5 levels in episodic angioedema and eosinophilia (Gleich's syndrome). AB - A patient with marked chronic eosinophilia and episodic angioedema (Gleich's syndrome) with a concomitant clonal proliferation of helper T-lymphocytes is described. This association has not previously been documented. The symptoms and clinical course differ markedly from the idiopathic hypereosinophilic syndrome, including an absence of end organ involvement. However, the disease appears to have a similar pathogenesis, with elevated levels of the eosinophil-specific cytokine interleukin-5 (IL-5) produced by the T-cell clone. PMID- 14565668 TI - Extramedullary acute myeloid leukemia (granulocytic sarcoma) with arm paresis, maculopapular exanthema and organ involvement. AB - Granulocytic sarcoma (extramedullary myelosarcoma, chloroma) is a rare extramedullary myeloid tumor which can occur at any anatomical site as isolated finding or associated with acute myelogenous leukemia (AML) or myelodysplastic syndrome (MDS). In this case, we describe a 71-year-old man who presented with incomplete paresis of the left arm, periorbital swelling, a maculopapular exanthema and organ involvement including testis and stomach. The tumors responded to combination chemotherapy and the patient fully recovered. However, after five months the patient relapsed and died quickly. This case confirms the importance of including granulocytic sarcoma in the differential diagnoses of a variety of diseases. In AML, the presence of granulocytic sarcoma is associated with worse overall survival. When diagnosed, it should be treated with intensive chemotherapy as soon as possible. PMID- 14565670 TI - Pulmonary embolism and thrombotic thrombocytopenic purpura in acute promyelocytic leukemia treated with all-trans retinoic acid. AB - We describe a patient with acute promyelocytic leukemia (APL) who developed pulmonary embolism (PE) and thrombotic thrombocytopenic purpura (TTP) during remission induction all-trans retinoic acid (ATRA) therapy. A 44-year-old man was diagnosed with APL and was treated with ATRA. On day 14, he developed PE, and on day 24, he developed TTP. Both PE and TTP occurred in association with leukocytosis due to ATRA administration. The PE responded to dexamethasone and TTP responded to plasma infusion. The PE and TTP remitted, and he achieved complete remission of APL. To our knowledge, there have been no reports of TTP occurring as a complication of ATRA therapy. PMID- 14565671 TI - Rapidly progressive human herpesvirus 8-associated solid anaplastic lymphoma in a patient with AIDS--associated Kaposi sarcoma. AB - We report a case of rapidly progressive solid lymphoma with anaplastic large cell morphology, followed by systemic Kaposi sarcoma in an adult patient with AIDS. The lymphoma cells expressed human herpesvirus 8 (HHV-8)-encoded latent and lytic proteins and Epstein-Barr virus-encoded small RNA, suggesting that this case could be categorized into HHV-8-associated solid lymphoma, a recently identified disease entity. PMID- 14565672 TI - Non-Hodgkin's lymphoma presenting as a giant chest wall mass. PMID- 14565673 TI - Rituximab in heavily pretreated cutaneous B-cell lymphoma. PMID- 14565674 TI - Idiopathic thrombocytopenic purpura and splenic marginal-zone B-cell lymphoma: a casual correlation? PMID- 14565675 TI - Proceedings of the International BiCONTACT Experts meeting. San Diego, California, USA. August 28, 2002. PMID- 14565676 TI - Fifteen years of experience with the BiCONTACT hip endoprosthesis system--the past, the present, the future. What has been achieved? AB - The BiCONTACT endoprosthesis system, in addition to having a distinctly improved material structure (hardness, lubricant couple, reduction of abrasions, etc.), demonstrates characteristics geared towards providing uniform, optimal treatment under economically favourable conditions for the widest possible stock of patients. Furthermore, it secures local stability and freedom from pain as early as possible, sparing or--even better--building bone with a biologically orientated implantation technique and obtaining correspondingly acceptable medium and long-term results via an appropriate surface coating for osseo-integration. The concept of the hip endoprosthesis, its design and implantation technique- which have not required modifications since its introduction in clinical practice 15 years ago--is capable of doing justice to all the individual, biological, instrumental and economic particularities mentioned. PMID- 14565677 TI - The periprosthetic bone remodelling process--signs of vital bone reaction. AB - Bone remodelling in cementless hip stem replacement occurs as interface and periprosthetic bone remodelling. Results of radiological, histological and bone mineral density (BMD) findings are discussed. Atrophic, hypertrophic and normotrophic bone reactions are described. Osteodensitometry measurements have shown a bone density decrease of 20% within the first 6 months in the region of the proximal femur. Measurements up to 4 years revealed no further loss of BMD. PMID- 14565678 TI - Long-term results with the BiCONTACT system--aspects to investigate and to learn from. AB - The BiCONTACT femoral stem for cementless fixation is being used without any technical modification after 15 years. The long-term results should be evaluated in this study. A consecutive series was continuously monitored in a prospective follow-up study. A survival analysis was performed, clinical results were rated according to the Harris score. There were 236 patients with 250 total hip replacements (THR); mean age at time of implantation was 58.2 years. Indications for THR included osteoarthritis (62.4%), dysplasia (16.8%), trauma (8.4%) and femoral-head necrosis (16.8%). Average time of follow-up evaluation was 8.9 years (range 7.4-10.7 years). At follow-up, 27 patients had died and two could not be located. Seven patients were revised--two for infection, one for recurrent dislocation, two for component undersizing with rapid subsidence, and one for aseptic loosening of a varus-malaligned stem; one radiologically well-fixed stem had been revised during acetabular revision. Survival estimate showed an overall survival rate of 97.1% after 11 years (confidence limits: 98.7% upper and 93.6% lower). Radiologically, tiny reactive lines (< 2 mm) were present in the distal zones of the femoral shaft, but no radiolucencies could be found in the proximal anchoring zone. Migration analysis with Ein-Bild-Rontgen-analyse/femoral component analysis (EBRA/FCA) demonstrated a very small amount of migration: in 31.0%, the overall migration was between 0.5 and 1 mm after 120 months; 8.5% had an absolute amount of subsidence exceeding 2 mm after 120 months (one case more than 3 mm). Mean subsidence was 0.2 mm after 3 months and 6 months, 0.3 mm after 12 months, and reached 0.5 mm after 10 years. An initial small amount of subsidence could be detected in 45.1%, and 15.5% had a late onset of subsidence. Continuous sinking could be found in 12.7%, while 26.8% had irregular patterns of migration. Clinical results were somewhat compromised by a higher-than-average rate of cup loosening (uncoated threaded cup). The average Harris hip score at follow-up was 84.3 points. Interestingly, no femoral osteolysis could be detected, even in cases with severe acetabular osteolyses, indicating sealing of the stem interface by tight osseointegration of the proximally-coated stem. PMID- 14565679 TI - Experiences with the Plasmacup--early stability, wear, remodelling, and outcome. AB - Whilst advances in cementing technique have led to improvement in the survival of cemented femoral stems in total hip arthroplasty (THA), cup failure due to aseptic loosening remains a major clinical problem. These observations have led to a move away from cemented cup designs, particularly in young patients, towards uncemented implants. The Plasmacup is a hemispherical, press-fit, cementless, titanium-shelled, acetabular component with a polyethylene liner. In this article we review our experience of its pattern of early migration, wear, bone remodelling, and mid-term survival. In 18 cups followed for 2 years in subjects with a mean age at operation of 58 years, the mean total vectorial cup migration was 0.75 mm, and cup orientation remained stable (EBRA method). The mean polyethylene linear wear rate over this period was 0.21 mm/year. In 27 cups followed for 6 months using dual-energy X-ray absorptiometry (DXA), average bone loss was -5%, and the pelvic bone-remodelling pattern was consistent with the rim loading principle of the cup design. In a clinical review of the outcome of 128 cups in 104 patients with a mean age at operation of 51 years and follow-up of 59 months, we found that 82% of patients had a good or very good Merle D'Aubigne score, and cup survival rate was 98% (Kaplan-Meier). Four cups had small radiographic areas of focal osteolysis and three had been revised (two for recurrent dislocation and one for deep sepsis; none were revised for aseptic loosening). The mean linear wear rate in this series was 0.14 mm/year. In conclusion, the Plasmacup shows satisfactory early stability, a wear rate similar to other uncemented cups, and favourable mid-term clinical function and survival rates. PMID- 14565680 TI - Plasmacup--special design. Aspects of the dysplastic acetabulum. AB - In Japan, the main reason for osteoarthritis of the hip joint is caused by dysplastic deformities. The implantation technique of cementless press-fit cup with multi-screw fixation and acetabular bone graft to reconstruct the true acetabular center is described. In addition, special design aspects of the BiCONTACT-N stem were optimized for these patients. Between January 1997 and December 1999, 29 patients (33 hips) received a BiCONTACT-N/Plasmacup-M total hip replacement. There were 28 women and one man. Average age at the time of THA of 64 (range 51-76) years. Average time of follow-up was 41 (range 25-59) months. According to the Crowe classification, 18 hips where classified as type 1 (less than 50% subluxation of the femoral head from the acetabulum), nine type 2 (50 75% subluxation), five type 3 (75-100% subluxation), and one type 4 (complete dislocation). At time of follow-up, there was no loosening of the acetabular implant or collapse of the acetabular bone graft. In the radiographic examination, 80% of the femoral implants showed spot welds in the transitional zone of the coated and uncoated area, and 96% sclerotic lines in the coated area. At the uncoated surface, radiolucent lines were seen in 12% and a cortical hypertrophy in 24%. No stress shielding could be observed. Stem subsidence was measured with an average value of 0.32 (range 0-1.6) mm. No patient complained about thigh pain, and no intraoperative complications were observed. Postoperative complications were two anterior dislocations due to cup malposition and excessive lumbar hypnosis. Three deep-venous thromboses where found with venography and treated with warfarin therapy. The limited experience indicates that BiCONTACT-N stem design in combination with the Plasmacup multihole implant is appropriate for the treatment of dysplastic hip deformities in Japanese patients. However, a long-term follow-up study is needed. PMID- 14565681 TI - Revision arthroplasty--femoral aspect: the concept to solve high grade defects. AB - The concept of and the results with a long-stem revision prosthesis based on the universal hip system BiCONTACT experiences are presented. After removal of the loosened hip stem via a transfemoral approach, the defect zone is bridged by a long-stem prosthesis with distal interlocking for higher primary stability. After bone healing and reconstruction thanks to the fracture healing mechanism, the interlocking bolts are removed and the implant can be stabilized again in the proximal part of the femur. In this area, osseointegration is supported by the microporous coating Plasmapore. The first 109 procedures with 70% high-grade femoral defects where presented for an average of 5.25 (maximum 9.7) years. Harris score improved from 45 to 75 points and implant survival rate was 85.3%. Restoration and reorganization of the proximal bone stock could be observed clinically, radiologically, and histologically. Evaluation of re-revisions showed suboptimal surgical procedures, poor bone structure, too early removal of interlocking bolts, and undersized implants as reasons for failure using this technique. We conclude that biological revision with the long BiCONTACT stem with distal interlocking option is a concept to solve problems associated with high bone-loss situations. PMID- 14565682 TI - Revision arthroplasty--acetabular aspect: cementless acetabular bone reconstruction. AB - The increasing occurrence of aseptic loosening, often accompanied by extensive acetabular defects, confronts the surgeon with severe difficulties. Multisegmental acetabular defects can be treated by bone grafting and metal antiprotrusion support rings. With these techniques, pelvis defects can be reconstructed to achieve a stable and permanent fixation of the acetabular component. Since 1983, more than 850 patients underwent revision surgery of the acetabular component using a reinforcement ring. Forty-one consecutive cases were available for follow-up, including clinical and radiographic examination as well as evaluation of patient history on average 7.3 years after operation. We report the follow-up results of these cases, which demonstrate that this kind of revision surgery guarantees a solid and stable situation at the acetabular component. PMID- 14565683 TI - OrthoPilot cup navigation--how to optimise cup positioning? AB - The kinematic cup navigation procedure is a surgically simple technique for improving the precision of cup placement. Without CT scans, a reduction of cup malpositions is possible. The rate of desired cup inclination and anteversion angles is markedly increased as shown by the first results of 147 navigated cups. While the inclination is reached precisely with a tendency to a slightly larger angle than desired, the postoperative anteversion angle shows a somewhat bigger variation than the intraoperatively navigated angle with a tendency to less anteversion. This might be due to a lack of information about the real pelvis position during X-ray control. CLINICAL RELEVANCE: dislocations of the prostheses might be reduced significantly. The kinematic cup navigation with the use of the OrthoPilot is an additional and useful tool for the surgeon. The procedure is simple, quick, and cost effective, does not cause specific complications, and does not require additional CT or MRI scans. Further improvement can be expected by additional hip stem navigation techniques in the near future. PMID- 14565684 TI - Image-based hip navigation. AB - After experimental and preclinical evaluation (HAP Paul Award 2001) of a CT-free image-guided surgical navigation system for acetabular cup placement, the system was introduced into clinical routine. The computation of the angular orientation of the cup is based on reference coordinates from the anterior pelvic plane (APP) concept. A hybrid strategy for pelvic landmark acquisition has been introduced involving percutaneous pointer-based digitization with the noninvasive biplanar landmark reconstruction using multiple registered fluoroscopy images. From January 2001 to May 2002, 118 consecutive patients (mean age 68 years, 82 male, 36 female, and 62 left and 56 right hip joints) were operated on with the hybrid CT-free navigation system. During each operation, the angular orientation of the inserted implant was recorded. To determine the placement accuracy of the acetabular components, the first 50 consecutive patients underwent a CT scan 7-10 days postoperatively to analyze the cup position relative to the APP. This was done blinded with commercial planning software. There was no significant learning curve observed for the use of the system. Mean values for postoperative inclination read 43 degrees (SD 3.0, range 37-49) and anteversion 19 degrees (SD 3.9, range 10-28). The resulting system accuracy, i.e., the difference between intraoperatively calculated cup orientation and postoperatively measured implant position, shows a maximum error of 5 degrees for the inclination (mean 1.5 degrees, SD 1.1) and 6 degrees for the anteversion (mean 2.4 degrees, SD 1.3). An accuracy of better than 5 degrees inclination and 6 degrees anteversion was achieved under clinical conditions, which implies that there is no significant difference in performance from the established CT-based navigation methods. Image guided CT-free cup navigation provides a reliable solution for future total hip arthroplasty (THA). PMID- 14565685 TI - Measuring subepithelial thickness using endobronchial ultrasonography in a patient with asthma: a case report. AB - The chronic inflammation of bronchial asthma is characterized by swelling of the subepithelial mucosa. However, it is difficult to assess subepithelial edema clinically. We report the case of a patient with asthma whose subepithelial edema was evaluated by endobronchial ultrasonography. Receiving montelukast 10 mg/day for 2 weeks, a 42-year-old man with mild, persistent asthma had his symptoms controlled by beta2-inhalation alone. Pretreatment endobronchial ultrasonography revealed subepithelial thickening in the right main stem bronchus, with a low absorption area suggestive of edema. Two weeks of montelukast therapy diminished the amount of subepithelial edema. Endobronchial ultrasonography is a promising technique for determining subepithelial edema in the asthmatic airway. PMID- 14565686 TI - Effects of plasmalemmal V-ATPase activity on plasma membrane potential of resident alveolar macrophages. AB - The acid-base status and functional responses of alveolar macrophages (mphi) are influenced by the activity of plasmalemmal V-type H+-pump (V-ATPase), an electrogenic H+ extruder that provides a possible link between intracellular pH (pHi) and plasma membrane potential (Em). This study examined the relationships among Em, pHi, and plasmalemmal V-ATPase activity in resident alveolar mphi from rabbits. Em and pHi were measured using fluorescent probes. Em was -46 mV and pHi was 7.14 at an extracellular pH (pHo) of 7.4. The pHi declined progressively at lower pHo values. Decrements in pHo, also caused depolarization of the plasma membrane, independent of V-ATPase activity. The pH effects on Em were sensitive to external K+, and hence, probably involved pH-sensitive K+ conductance. H+ were not distributed at equilibrium across the plasma membrane. V-ATPase activity was a major determinant of the transmembrane H+ disequilibrium. Pump inhibition with bafilomycin A1 caused cytosolic acidification, due most likely to the retention of metabolically generated H+. V-ATPase inhibition also caused depolarization of the plasma membrane, but the effects were mediated indirectly via the accompanying pHi changes. V-ATPase activity was sensitive to Em. Em hyperpolarization (valinomycin-clamp) reduced V-ATPase activity, causing an acidic shift in baseline pHi under steady-state conditions and slowing pHi recovery from NH4Cl prepulse acid-loads. The findings indicate that a complex relationship exists among Em, pHi, and pHo that was partially mediated by plasmalemmal V-ATPase activity. This relationship could have important consequences for the expression of pH- and/or voltage-sensitive functions in alveolar mphi. PMID- 14565687 TI - Prenatal nicotine exposure and pulmonary barotrauma of newborns. AB - Smoking in adults increases the relative risk of contracting spontaneous pneumothorax, a form of pulmonary barotrauma. Maternal smoking habits affect the fetus. Pregnant females attend the antenatal clinic at the 8th to 12th weeks of pregnancy. There is a participation rate of 99% of all births in Sweden. Their smoking habits were registered at this stage. This study supported the hypothesis that the registered maternal smoking habits covariated with the risk of contracting pulmonary barotrauma in newborn infants. The infants of smokers do not seem to be at higher risk (95% C.I. of RR: 0.78-0.99) for contracting pulmonary barotrauma than those of nonsmokers. Thus far, the hypothesis is even rejected at the 5% significance level. However, after considering other factors, especially mother's education, it seems to be an open question whether or not a weak covariation is present. Newborn boys run almost twice the risk of contracting pulmonary barotrauma than girls. Furthermore, we found an increased risk for contracting pulmonary barotrauma in the subcohort of newborns whose mothers' smoking habits were not reported. PMID- 14565688 TI - The effects of one-week fluticasone propionate inhalation therapy for Tc-99m DTPA radioaerosol distribution in asthma of children: a preliminary report. AB - This study evaluated the effects of fluticasone propionate inhalation therapy for the distribution pattern of Tc-99m DTPA radioaerosols in 10 children with asthma. The homogeneous degree of depositing Tc-99m DTPA radioaerosol was evaluated using a modified standard score system over the bilateral lungs. The baseline scores were calculated from Tc-99m DTPA radioaerosol inhalation lung scintigraphy before inhalation therapy (100 microg fluticasone propionate two times daily for one week), and the scores were recalculated after inhalation therapy to evaluate the effects of one-week of fluticasone propionate inhalation therapy for Tc-99m DTPA radioaerosol distribution patterns. After one week of fluticasone propionate inhalation therapy, the scores were decreased in all of the 10 children, which may mean that the bronchial constriction degree due to asthma is decreased. In addition, there was a significantly statistical difference in the scores before and after one-week fluticasone propionate inhalation therapy (p < 0.05). In conclusion, one-week fluticasone propionate inhalation therapy could significantly improve the bronchial constriction due to asthma in children based on the evidence of Tc-99m DTPA radioaerosol inhalation lung scintigraphic findings. PMID- 14565689 TI - Pulmonary hemodynamics in obstructive sleep apnea: frequency and causes of pulmonary hypertension. AB - The association between nocturnal apneas and transient pulmonary hypertension (PHT) has been well documented. However, there is controversy over the frequency and pathophysiological mechanisms of daytime pulmonary hypertension in patients with obstructive sleep apnea (OSAS). The present study sought to evaluate frequency and mechanisms of pulmonary hypertension in patients with OSAS. It included 49 consecutive patients with polysomnographically proven OSAS without pathological lung function testing. All patients performed daytime measurements of pulmonary hemodynamics at rest and during exercise (50-75W). Six patients (12%) had resting PHT mean pulmonary of artery pressure (PAPM) of >20 mmHg), whereas 39 patients (80%) showed PHT during exercise (PAPM >30 mmHg). Multiple regression analysis revealed 3 independent contributing factors for mean pulmonary artery pressure during exercise (PAPMmax): body mass index, age and total lung capacity % of predicted. Twenty-five of the 39 patients with pathologically high PAPMmax (64%) showed elevated pulmonary capillary wedge pressures (PCWPmax > 20 mmHg), whereas no patient had elevated pulmonary vascular resistance (PVRmax > 120 dynes x s x cm(-5)). In conclusion, daytime PHT during exercise is frequently seen in patients with OSAS and normal lung function testing and is mainly caused by abnormally high PCWP, whereas PVR seems to play a minor role. PMID- 14565690 TI - Practitioner perspectives on the role of science in environmental impact assessment. AB - A large body of literature addresses the role of science in environmental impact assessment (EIA) but less attention has been given to the views of practitioners themselves. In this research a survey of 31 EIA practitioners in Western Australia was undertaken to determine their perceptions of the quality and importance of science in EIA. The survey results are compared with previous theoretical, empirical, and survey studies of the role of science in EIA. Interview questions addressed the role of science in impact prediction, monitoring activities, mitigation and management, and EIA decision-making. It was clear from the interviews that many practitioners are satisfied with the quality of science currently used in EIA, but do not believe that it is given sufficient importance in the process. The quality and importance of science in the predecision stages of EIA was rated higher than in the postdecision stages. While science was perceived to provide the basis for baseline data collection, impact prediction, and mitigation design, it was seen to be less important during decision-making and ongoing project management. Science was seen to be just one input to decision-makers along with other factors such as sociopolitical and economic considerations. While time and budget constraints were seen to limit the scientific integrity of EIA activities, pressure from the public and regulatory authorities increased it. Improving the scientific component of EIA will require consideration of all these factors, not just the technical issues. PMID- 14565691 TI - Landscape changes in the Mullica River basin of the Pinelands National Reserve, New Jersey, USA. AB - In 1979, the Pinelands Commission was established as a regional land-use planning and regulatory agency charged with the implementation of a Comprehensive Management Plan (CMP) for the Pinelands National Reserve (New Jersey, USA). The CMP was created to balance land preservation and development interests in the Reserve. Because water-quality degradation from developed and agricultural landscapes is associated with changes in the composition of biological communities, monitoring landscape changes provides one of the most direct measures of the impact of land-use policies on the Pinelands ecosystem. In this study, we prepared detailed, land-cover maps within randomly selected aerial photograph plots for a major watershed in the Reserve. We used these land-cover maps to quantify changes in landscape composition and structure (i.e., patch size, patch area, and number of patches) and characterize land-cover transitions in the basin between 1979 and 1991. Because the study period represented the first 12 years of the regional-planning effort in the Reserve, we evaluated the relationship between land-cover transitions and Commission management-area designations which permit different land-use intensities. Although the landscape composition was similar in 1979 and 1991, we found an increase in the total area and number of developed-land, managed-grassland, and barren-land patches. An increase in the number of patches and a decrease in the total area and median and third-quartile patch sizes for forest land and for all patches regardless of cover type indicated that fragmentation of forest land and the landscape as a whole occurred during the study period. The major land-cover transitions that occurred during the period were the loss of forest land to development and associated cover types and the conversion of one agricultural type to another. Overall, land-cover transitions during the period were found to be consistent with the Commission management-area designations, which indicated that the regional-planning effort has been successful in directing human activities to appropriate areas of the basin. PMID- 14565692 TI - Forest perception and knowledge of hikers and mountain bikers in two different areas in northwestern Switzerland. AB - Recreational activities can have major impacts on vegetation and wildlife in frequently visited forests. We assessed forest perception and knowledge (state, functions, and species diversity) among hikers and mountain bikers in a frequently visited, seminatural suburban recreation forest (Muttenz) and in a more distantly situated, naturally grown excursion forest (Wasserfallen) in northwestern Switzerland. In all, 239 hikers and 126 mountain bikers were interviewed. Mountain bikers in both forests and hikers in the more intensely used recreation forest at Muttenz assessed the state of the forest less optimistically and showed a higher awareness of the negative impact of recreational activities on the flora and fauna than hikers at Wasserfallen. Furthermore, mountain bikers seemed aware of the social conflicts caused by their activity, since they appreciated neutral or positive encounters with other forest visitors. In contrast, 57% of hikers at Muttenz reported on negative experiences with other forest visitors, particularly with mountain bikers. In general, the interviewees' ecological and biological forest knowledge (forest type and function, species diversity) was rather high. A large proportion was aware of the pros and cons of different forest conditions for plants and animals, and could name or recognize at least some plant and/or animal species typical for the visited forest. The forest knowledge was neither influenced by the type of recreational activity carried out nor by any aspect of forest visit behavior (frequency and duration of forest visits, means of transportation and travel distance to forest). However, the interviewees' forest knowledge was influenced by their age and educational level. PMID- 14565693 TI - Impacts to water quality and fish habitat associated with maintaining natural channels for flood control. AB - Field investigation conducted on Boulder Creek in Boulder, Colorado evaluated impacts of flood control maintenance activities on flood conveyance, water quality, and fish habitat. Thirty-nine transects were monitored at one control site and two maintenance sites over a period of eight months. Each site was visited on more than 50 occasions in order to characterize pre- and post maintenance conditions, and to monitor maintenance activities. Measurements along the transects included substrate composition, flow depth, velocity, and elevation. Reach-average values were assigned to variables such as in-stream vegetation, streambank stability, and woody vegetation before and after maintenance. Water temperature, dissolved oxygen, pH, specific conductance, and turbidity were sampled, and habitat suitability indices were developed pre- and post-maintenance for seven indicator fish species. Water quality impacts during maintenance consisted of high turbidity levels (> 400 NTU), which retumed to background levels (0.1-15 NTU) overnight, as well as changes in mean temperature and pH. Alteration of physical channel characteristics as a result of maintenance had limited effects on habitat quality for four of seven fish species, but caused improvements in habitat quality for three fish species. The main implications of this study for floodplain management are that: (1) Flood control maintenance practices can be in direct conflict with water quality and fish habitat objectives, and should be carefully designed and implemented by an interdisciplinary team. (2) Physical habitat for some fish species can be improved as well as reduced by maintenance activities. Habitat suitability curves may be useful tools for evaluating limiting factors of the habitat and for identifying opportunities for habitat improvements as part of maintenance. PMID- 14565694 TI - Bioremediation of soil degraded by sewage sludge: effects on soil properties and erosion losses. AB - Soils in the Mediterranean area are very prone to erosion due to the loss of organic matter and the consequent lack of protective vegetation. In this experiment a Mediterranean degraded soil with a 15% slope was amended at a rate of 250 t ha(-1) wet weight with sewage sludge and with a mixture of sewage sludge and barley straw (70% carbon from sewage sludge and 30% from the straw) in order to study their influence on soil structure recovery and hence the soils's resistance to erosion processes. Both types of organic amendment led to an improvement in several soil properties (physical, biological, and microbiological) as a result of the spontaneous growth plant covering that became evident three months after amendment. This vegetation remained throughout the two years of the experiment and prevented the water erosion processes that normally precede soil degradation. Amendment by sewage sludge alone reduced soil loss by 80% compared with the control soil, while the mixture that included both sewage sludge and barley straw reduced losses by 84%, both reducing runoff by 57%. The amended soils showed increases in the percentage of stable aggregates, the levels of the total and water-soluble C fractions, microbial biomass C, basal respiration, and the activity of the different enzymes involved in the biogeochemical cycles of C, N, and P. The results confirm the usefulness of sewage sludge as an organic amendment for recovering damaged soils. PMID- 14565695 TI - Effects of soil conditions on survival and growth of black willow cuttings. AB - Current streambank restoration efforts focus on providing bank stability, enhancing water quality, and improving woody habitat using native vegetation rather than traditional engineering techniques. However, in most cases harsh site conditions limit restoration success. A two-year field study was conducted at Twentymile Creek, in northern Mississippi, investigating edaphic factors governing the survival of black willow (Salix nigra) cuttings used for streambank restoration. Low height growth, above-ground biomass production, and average leaf area were observed in willow cuttings grown in plots subjected to moisture deficits. However, sediment texture emerged as the dominant factor determining willow post growth, health, and survival. Shoot biomass, leaf biomass, and total above-ground biomass were 15-, 10-, and 14-fold greater for large willow cuttings (posts) grown in plots with sandy sediments relative to those grown in plots with similar moisture and soil redox potential but with silt and clay sediments. Average leaf size, average leaf mass and specific leaf area were all lower in fine textured plots. Under moisture conditions present at our sites, coarse grained sediment (sand) was more conducive to willow growth, biomass production, and survival than were fine-grained sediments (silt/clay). Our results strongly suggest that soil texture and moisture conditions can determine restoration success. Therefore, it is critical that site conditions are factored into the selection of project locations prior to the initiation of willow planting restoration projects. PMID- 14565696 TI - Geographic analysis of forest health indicators using spatial scan statistics. AB - Geographically explicit analysis tools are needed to assess forest health indicators that are measured over large regions. Spatial scan statistics can be used to detect spatial or spatiotemporal clusters of forests representing hotspots of extreme indicator values. This paper demonstrates the approach through analyses of forest fragmentation indicators in the southeastern United States and insect and pathogen indicators in the Pacific Northwest United States. The scan statistic detected four spatial clusters of fragmented forest including a hotspot in the Piedmont and Coastal Plain region. Three recurring clusters of insect and pathogen occurrence were found in the Pacific Northwest. Spatial scan statistics are a powerful new tool that can be used to identify potential forest health problems. PMID- 14565697 TI - Roadside measurements of deforestation in the Amazon area of Bolivia. AB - This study describes the deforestation impact from three settlements, 5 de Junio, El Tigre, and 30 de Agosto, near the Madidi National Park in Bolivia. First deforestation rates along the road that connects the settlements and then deforestation within each settlement are examined. Landsat Thematic Mapper (TM) imagery from 1987, 1997, and 2000 were used to measure the deforestation. Face-to face interviews were also used to collect detailed social and land-use information. Road-building and farming have been the major causes of deforestation in this region. The TM measurements indicate a high increase in the deforestation rate along the road in the area of study. The results also show a rapidly escalating deforestation rate in 5 de Junio, a potential high future deforestation rate in El Tigre, and a gradual deforestation increase in 30 de Agosto. The information provided by the settlers about clearing was compared with the imagery analysis. Generally, the settler's estimates were lower than the deforestation as derived from the imagery. PMID- 14565698 TI - Environmental management systems at the industrial park level in China. AB - Environmental management systems (EMSs), such as International Organization for Standardization (ISO) 14001, can be used as a tool in China by industrial park managers to improve their environmental performance. This article uses the case of the Dalian Economic and Technological Development Zone (DETDZ) to show how to establish a comprehensive environmental management system (CEMS) according to the ISO 14001 standard at the industrial park level by considering local realities. The particularly interesting feature of this case study is the use of a CEMS (in this case, ISO 14001) by the administrative group of the DETDZ to develop a more comprehensive approach to the wide range of environmental issues that they face in running the zone. In essence the goal is to address many of the issues at the level of the zone. The incentives, benefits, and barriers associated with implementing ISO 14001 are described. However, implementation of an EMS should not be thought of as the ultimate objective for an industrial park's environmental management. The next steps include encouraging further public participation and taking an integrated approach leading to an industrial ecosystem, which can realize better environmental performance at the industrial park level. PMID- 14565699 TI - Integrating bioassessment and ecological risk assessment: an approach to developing numerical water-quality criteria. AB - Ioassessment is used worldwide to monitor aquatic health but is infrequently used with risk-assessment objectives, such as supporting the development of defensible, numerical water-quality criteria. To this end, we present a generalized approach for detecting potential ecological thresholds using assemblage-level attributes and a multimetric index (Index of Biological Integrity-IBI) as endpoints in response to numerical changes in water quality. To illustrate the approach, we used existing macroinvertebrate and surface-water total phosphorus (TP) datasets from an observed P gradient and a P-dosing experiment in wetlands of the south Florida coastal plain nutrient ecoregion. Ten assemblage attributes were identified as potential metrics using the observational data, and five were validated in the experiment. These five core metrics were subjected individually and as an aggregated Nutrient-IBI to nonparametric changepoint analysis (nCPA) to estimate cumulative probabilities of a threshold response to TP. Threshold responses were evident for all metrics and the IBI, and were repeatable through time. Results from the observed gradient indicated that a threshold was > or = 50% probable between 12.6 and 19.4 microg/L TP for individual metrics and 14.8 microg/L TP for the IBI. Results from the P dosing experiment revealed > or = 50% probability of a response between 11.2 and 13.0 microg/L TP for the metrics and 12.3 microg/L TP for the IBI. Uncertainty analysis indicated a low (typically > or = 5%) probability that an IBI threshold occurred at < or = 10 microg/L TP, while there was > or = 95% certainty that the threshold was < or = 17 microg/L TP. The weight-of-evidence produced from these analyses implies that a TP concentration > 12-15 microg/L is likely to cause degradation of macroinvertebrate assemblage structure and function, a reflection of biological integrity, in the study area. This finding may assist in the development of a numerical water-quality criterion for TP in this ecoregion, and illustrates the utility of bioassessment to environmental decision-making. PMID- 14565700 TI - Vegetation composition, dynamics, and management of a bracken-grassland and northern-dry forest ecosystem. AB - We investigated differences in vegetation composition and dynamics for two globally rare ecosystems, bracken-grasslands and northern-dry forests of northern Wisconsin. These ecosystems commonly have been viewed as degraded pine barrens. Bracken-grasslands contained a high dominance of exotic species, low native richness, and no obvious prairie species, suggesting logging-era anthropogenic origins. Differences in cover for common plants among ecosystems were examined using Mann-Whitney U tests of equivalence. Cover of all 8 graminoid species, 4 of 5 Ericaceae and Myricaceae species, and 10 of 17 species of forbs were significantly different between ecosystems. Vegetation changes over a 4-year period were examined through detrended correspondence analysis (DCA) and analysis of variance (ANOVA) repeated measures. DCA analyses of community composition failed to detect significant temporal trends within individual management units, although differences were apparent between ecosystems, regardless of sample year. In addition, no apparent patterns could be detected between years when comparing dominant individual species to management history (prescribed fire). This is contrary to what would be expected for a degraded pine barrens and questions the efficacy of using repeated prescribed fire as a management tool in bracken grasslands. Methods for conservation and restoration of xeric ecosystems of northern Wisconsin have historically relied heavily on single species (e.g., sharp-tailed grouse) wildlife models, without full consideration of other factors. We suggest that stakeholders involved in these restoration projects examine historic processes and reference conditions prior to formulating management goals. Greater attention to the differentiation and individual management needs of pine barrens, northern-dry forests, and bracken-grasslands is needed. PMID- 14565701 TI - Self-organizing maps for integrated environmental assessment of the Mid-Atlantic region. AB - A new method has been developed to perform environmental assessment at regional scale. This involves a combination of a self-organizing map (SOM) neural network and principal component analysis (PCA). The method is capable of clustering ecosystems in terms of environmental conditions and suggesting relative cumulative environmental impacts of multiple factors across a large region. Using data on land-cover, population, roads, streams, air pollution, and topography of the Mid-Atlantic region, the method was able to indicate areas that are in relatively poor environmental condition or vulnerable to future deterioration. Combining the strengths of SOM with those of PCA, the method offers an easy and useful way to perform a regional environmental assessment. Compared with traditional clustering and ranking approaches, the described method has considerable advantages, such as providing a valuable means for visualizing complex multidimensional environmental data at multiple scales and offering a single assessment or ranking needed for a regional environmental assessment while still facilitating the opportunity for more detailed analyses. PMID- 14565702 TI - The anatomy of the mitral valve: a retrospective analysis of yesterday's future. PMID- 14565703 TI - Attitudinally correct designation of papillary muscles. PMID- 14565704 TI - Echocardiographic roadmap of the mitral valve anatomy. PMID- 14565705 TI - The subcoronary technique versus the root technique for autograft surgery. PMID- 14565706 TI - Insights from 36 years' follow up of a patient with the Ross operation. AB - Pulmonary autograft replacement of the aortic valve has been shown to have many potential advantages over other forms of aortic valve replacement. However, there is still a paucity of studies that provide new information about the possible behavior of the two valves and the cardiac and extracardiac responses in the very long-term. Here, a patient is presented who underwent this operation 36 years ago, together with detailed anatomical and physiological documentation. This case report serves to illustrate several important features relating to the late behavior of the two valves, the response of the two ventricles, and the 'quality of life' of the patient. PMID- 14565707 TI - Evidence for rheumatic valve disease in patients with severe tricuspid regurgitation long after mitral valve surgery: the role of 3D echo reconstruction. AB - BACKGROUND AND AIM OF THE STUDY: Although severe tricuspid regurgitation (TR) is a well-recognized, long-term complication of rheumatic mitral valve replacement that impairs the functional results of surgery, its exact basis remains unclear and its management is unsatisfactory. The study aim was to obtain a detailed assessment of tricuspid valve morphology and function using 2D transesophageal echocardiography (TEE) with 3D reconstruction, and to determine long-term clinical outcome in patients after surgery for rheumatic mitral valve disease. METHODS: A total of 42 patients (mean age 50 +/- 10 years) was followed up; 39 patients had mitral replacement and three had valvotomy. Thirty patients had developed impaired exercise tolerance, fluid retention and echocardiographic evidence of severe TR at 8.2 +/- 2.6 years after surgery; the remainder had mild regurgitation. RESULTS: Follow up showed greater mortality in the severe TR group, with approximately 50% survival at 60 months after diagnosis compared with mild TR. None of the patients with severe TR had a dysfunctional mitral prosthesis. In these patients, transthoracic echo-Doppler showed enlarged right atrium and right ventricle, a mean transtricuspid retrograde pressure drop of 15 +/- 4 mmHg and apparently normal leaflet anatomy. Twenty patients (15 with severe TR) underwent a TEE and 3D reconstruction study for further evaluation. Abnormal leaflet anatomy was demonstrated in all patients with severe TR, with restricted leaflet motion in 10, leaflet shortening and thickening in the remainder, and dilatation of tricuspid valve annular insertion suggestive of rheumatic involvement. Although diastolic transtricuspid velocities were increased (peak flow 0.8 +/- 0.1 m/s) in these patients due to increased stroke volume, significant tricuspid stenosis was present in only two cases (mean gradient 4 and 3 mmHg respectively). Histopathology confirmed the presence of leaflet vascularization and extensive fibrosis in two patients who underwent tricuspid valve replacement. CONCLUSION: Rheumatic leaflet involvement contributes to severe TR occurring long after mitral valve replacement, though overt stenosis is uncommon. Knowledge of the structural basis of this condition may thus improve its long-term management, possibly with early tricuspid valve repair. PMID- 14565708 TI - Current profile of acute rheumatic fever and valvulitis in southern India. AB - BACKGROUND AND AIM OF THE STUDY: Rheumatic fever (RF) incidence has declined dramatically in the West, but still accounts for a major percentage of cardiac debility in developing countries. The study aim was to analyze the incidence, clinical profile and echocardiography findings in acute RF over the past decade. METHODS: The records of acute RF admissions to a tertiary care medical college hospital in southern India during the past 10 years were analyzed retrospectively. Chronic rheumatic heart disease and recurrence of RF were excluded from the study. Patients with a first episode of acute RF were studied only if the case records were available, and basic laboratory and echocardiography studies were completed. Thus, a total of 163 patients (81 males, 82 females) qualified for the study. RESULTS: There was no significant decline in the annual incidence of the first episode of acute RE The most common presenting symptoms were arthralgiaarthritis (n = 157), fever (n = 137) and breathlessness (n = 81). Carditis (67.5%) and arthritis (44.2%) were the most often-reported Jones criteria. Trivial to mild mitral regurgitation by echocardiography occurred in 81%, followed by significant mitral regurgitation in 5.5% and mild aortic regurgitation in 25%. The 'youngest' and 'oldest' first episodes of RF were recorded at ages of 2 and 29 years, respectively. CONCLUSION: The incidence of RF in India does not show the declining trends of the Western world. The application of Jones criteria for diagnosis remains relevant, though echocardiography is increasingly called upon to 'confirm' clinical diagnosis and help manage these patients in an appropriate manner. PMID- 14565709 TI - Clinical spectrum of chronic rheumatic heart disease in India. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to determine prevalence and patterns of chronic rheumatic heart disease (RHD) in developing countries, where it remains a major cause of mortality and morbidity. The incidence of different valvular lesions and complications in chronic RHD were analyzed. METHODS: The study design was a retrospective case series analysis in the setting of a tertiary care institution in southern India. Participants were consecutive patients registered under 'chronic RHD' in the cardiology department of the authors' institution over the past 20 years. Data are presented for 10,000 cases in two age groups: group I, aged < or = 18 years (n = 2,910); and group II, aged > 18 years (n = 7,090). RESULTS: Mitral regurgitation was the single most common lesion (n = 1,007) in group I, while the dominant lesion in group II was mitral stenosis (n = 2,943). Isolated aortic valve disease was seen in 130 (4.5%) and 195 (2.8%) cases in groups I and II, respectively. Tricuspid stenosis was seen in 45 cases, and rheumatic involvement of all four cardiac valves was documented in four cases. Pulmonary hypertension was present in 42.4% and 80.8% in groups I and II, respectively, and functional tricuspid regurgitation in 38.9% and 77.2%, respectively. Overall, 5.9% of patients had atrial fibrillation, 0.9% had left atrial thrombus (seen on transthoracic echocardiography) and 0.4% had embolic cerebrovascular events. Pericardial effusion was present in 0.7% cases, and infective endocarditis was noted at presentation in 0.6%. CONCLUSION: Chronic RHD in developing countries is associated with major complications and high mortality. The critical evaluation of individual lesions must be combined with frequent overall clinical evaluation in order to time appropriate medical and surgical interventions. PMID- 14565710 TI - Closed mitral commissurotomy: a most useful operation in the past. Is it justified in the present and does it have a future? PMID- 14565711 TI - The place of closed mitral valvotomy in the modern cardiac surgery era. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to investigate the long-term efficacy of closed mitral valvotomy (CMV). METHODS: Data obtained over a 36-year period from 1,134 patients who underwent CMV were analyzed. The analysis was carried out retrospectively from hospital records, with follow up examinations being conducted mainly at the outpatient clinic. RESULTS: In-hospital mortality (< or = 30 days after surgery) was 0.4% (n = 5, all closed procedures). Cardiac failure was the main cause of early death, and postoperative peripheral embolism occurred in five cases (0.5%). Freedom from thromboembolism was 99.0 +/- 0.5% at 36 years. Operative results were satisfactory in most patients, and severe mitral incompetence was seen only in three cases. Post-valvotomy mitral regurgitation occurred in 88 patients (7.7%) during the first year after CMV. Reoperation was performed in 500 patients (44.1%). The mean interval between CMV and reoperation was 141.1 +/- 80.8 months (range: 1-436 months). Fourteen patients were reoperated on for mitral regurgitation, 485 for mitral restenosis, and five for mixed mitral valve disease (stenosis and regurgitation). Freedom from reoperation after CMV was 81.4 +/- 1.3% at 10 years, 16.4 +/- 2.1% at 20 years, 3.1 +/- 1.2% at 20 years, and 0% at 36 years. Cox regression analysis indicated that impaired functional capacity, reduced mitral valve area, gradual increase in left atrial diameter and postoperative mitral insufficiency increased the reoperation rate after CMV. CONCLUSION: When compared with percutaneous balloon or surgical open valvotomy, CMV represents a satisfactory technique in terms of simplicity, high efficacy and lower cost. PMID- 14565713 TI - Role of inadequate adaptive left ventricular hypertrophy in the genesis of mitral regurgitation in patients with severe aortic stenosis: implications for its prevention. AB - BACKGROUND AND AIM OF THE STUDY: Mitral regurgitation (MR) is frequent in patients with severe calcific aortic stenosis (AS). This complicates not only the clinical course of AS, but also its surgical management. The aim of the present study was to investigate the mechanism of genesis of MR in patients with severe AS. METHODS: The echocardiographic database was searched for subjects with severe AS defined as a calculated (continuity equation) aortic valve area < 0.7 cm2. Patients with previous valve surgery were excluded; thus, the study group comprised 123 patients. RESULTS: Among 123 patients (mean age 75 +/- 10 years) with severe AS, 54 (44%) had no MR, 37 (30%) had mild MR, 20 (16%) had moderate MR, and 12 (10%) had severe MR. Hence, moderate or severe MR was present in approximately 25% of patients. Patients with moderate or severe MR had a larger left ventricular (LV) end-diastolic diameter (5.1 +/- 1.0 versus 4.8 +/- 0.8 cm; p = 0.08), larger LV end-systolic diameter (3.8 +/- 1.2 versus 3.1 +/- 0.8 cm; p = 0.001), lower LV ejection fraction (40 +/- 16 versus 58 +/- 18%; p = 0.0001), higher degree of aortic regurgitation (p = 0.002), larger left atrial diameter (4.7 +/- 0.9 versus 4.1 +/- 0.6 cm; p = 0.001), lower LV free wall thickness (1.1 +/- 0.2 versus 1.3 +/- 0.4 cm; p = 0.05), and lower combined wall thickness (2.4 +/- 0.3 versus 2.7 +/- 0.5 cm; p = 0.02) and relative wall thickness (0.5 +/- 0.1 versus 0.6 +/- 0.1 cm; p = 0.02). Both groups had similar degrees of AS and mitral annular calcification. CONCLUSION: MR in severe AS is associated with a larger LV size and lesser wall thickness, and this may result from failure of adequate adaptive LV hypertrophy necessitated by the pressure overload imposed by AS. This might have important clinical implications in terms of timing of aortic valve replacement before the left ventricle begins to dilate, and also in the choice of pharmacologic therapy that may modulate the adaptive response of the left ventricle. PMID- 14565712 TI - Remodeling of the cardiac extracellular matrix differs between volume- and pressure-overloaded ventricles and is specific for each heart valve lesion. AB - BACKGROUND AND AIM OF THE STUDY: Different patterns of fibroblast-mediated remodeling of the extracellular matrix (ECM) might be expected between patients with pressure- and volume-induced left ventricular hypertrophy. METHODS: Patients with chronic pressure-overload due to aortic stenosis (AS, n = 19) and chronic volume-overload due to either aortic regurgitation (AR, n = 9) or mitral regurgitation (MR, n = 10) were examined. The amount of interstitial collagen was quantified by using circular-polarization-microscopy in picrosirius red-stained biopsies. Biopsies from 10 patients with mild cardiomyopathy served as controls. In a subgroup, collagen type-I (Coll-I) and collagen type-III (Coll-III) gene expression was evaluated by quantitative RT-PCR. After immunohistological staining, procollagen-I/procollagen-III ratio and density of fibroblasts (FB) per vision-field were analyzed. RESULTS: The amount of interstitial cardiac fibrosis (ICF) was significantly higher in AS (5.7 +/- 4.1 g/m2), AR (8.8 +/- 4.9 g/m2) or MR (4.7 +/- 2.8 g/m2) than in controls (2.3 +/- 1.5 g/m2) (p = 0.003). The amount of thick collagen fibers was higher in AR than in AS, where-by density of fibroblasts did not differ. In volume-overloaded ventricles, the Coll-I/Coll-III ratio was shifted towards Coll-I, and in pressure-overloaded ventricles towards Coll-III. The severity of ECM remodeling was positively correlated with wall stress and impaired diastolic function, but not with systolic function or clinical symptoms. CONCLUSION: Remodeling of the ECM is specific for left ventricular volume and pressure overload, and may serve as an early indicator for inadequate adaptation. PMID- 14565714 TI - Expression and function of the integrin alpha9beta1 in bovine aortic valve interstitial cells. AB - BACKGROUND AND AIM OF THE STUDY: Aortic valve interstitial cells (VIC), the most prevalent valve leaflet cells, have not been well studied. However, recent interest in constructing tissue-engineered living heart valves has provided motivation to further an understanding of the molecular and cellular biology of VIC. Since cell-extracellular matrix interactions are critical for tissue morphogenesis, adhesive interactions and integrin function in VIC were investigated. METHODS: Bovine and baboon VIC were isolated and characterized by morphology, cytoskeletal protein expression and integrin expression. The interaction of VIC integrin alpha9beta1 with osteopontin, a ligand known to be up regulated in the wounded valve, was examined. The ability of bovine VIC alpha9beta1 to mediate adhesion, focal contact formation, migration and proliferation when plated on the immobilized osteopontin substrates, was investigated. RESULTS: Cultured bovine VIC possessed characteristics of both fibroblasts and smooth muscle cells. Like fibroblasts, bovine VIC had an elongated morphology with long cytoplasmic processes. Like smooth muscle cells, bovine VIC expressed alpha-smooth muscle actin and SM22alpha. Using fluorescent flow cytometry, the surface expression of integrins alpha9beta1 and alpha(v)beta3 in bovine VIC and integrins alpha1beta1, alpha2beta1, and alpha5beta1 in baboon VIC was identified. Expression of alpha9beta1 in cultured VIC and in native valve leaflets was further confirmed by Western blot and RT-PCR analysis. Bovine VIC were found to adhere to the 30N fragment of osteopontin in an alpha9beta1 dependent manner, and this interaction was mediated through the SVVYGLR motif. Bovine VIC also formed focal contacts through the alpha9beta1 integrin, but osteopontin (30N) did not stimulate migration or proliferation of bovine VIC through the integrin alpha9beta1. CONCLUSION: Integrin alpha9beta1 appears to be predominantly involved in controlling anchorage rather than movement or proliferation, in bovine VIC. For tissue engineering, knowledge of the VIC integrin profile can be applied to the rational design of scaffolds with the appropriate ligands for tissue formation. The VIC a9beta1-SVVYGLR interaction could be exploited to promote cell anchorage within a tissue engineering scaffold. PMID- 14565715 TI - Human pulmonary valve endothelial cells express functional adhesion molecules for leukocytes. AB - BACKGROUND AND AIM OF THE STUDY: Histopathological studies of rejected orthotopic heart transplants suggest that cardiac valve endothelium is spared the inflammatory cell infiltration and tissue damage that occurs in the myocardium. To test whether this apparent protection from leukocyte invasion might be an inherent feature of the valve endothelium, leukocyte adhesion molecule expression and function were analyzed in human pulmonary valve endothelial cells (HPVEC). Use of cultured HPVEC allowed delineation of the potential contribution of functional adhesion molecules from the contribution of hemodynamic forces exerted on the leaflet surface in vivo METHODS AND RESULTS: HPVEC express E-selectin, ICAM-1, and VCAM-1 in response to the inflammatory cytokine tumor necrosis factor alpha (TNF-alpha) similarly to other types of cultured human endothelial cells. In a static cell adhesion assay, E-selectin-mediated adhesion of HL-60 cells, a human promyelocytic leukemia cell line, and U937 cells, a human monocytic cell line, was determined in cells treated with TNF-alpha for 5 h. After 24 h of TNF alpha, adhesion of U937 cells to HPVEC was mediated primarily by VCAM-1, consistent with the high expression of VCAM-1 and diminished expression of E selectin at 24 h. CONCLUSION: These results demonstrate that HPVEC express functional leukocyte adhesion molecules in vitro and suggest that cardiac valve endothelium is competent to initiate leukocyte adhesion. Thus, other factors, such as the hemodynamic forces exerted on the valve, may contribute to the apparent protection from inflammatory cell infiltration in vivo. PMID- 14565716 TI - In-vitro endothelialization of patches of bovine jugular vein stabilized by dye mediated photooxidation. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to evaluate the cytotoxicity of photooxidatively stabilized bovine jugular vein and the in-vitro endothelialization properties of such material. METHODS: Bovine jugular veins were cross-linked by dye-mediated photooxidation and cut into 4 cm2 pieces. Ovine jugular vein endothelial cells were isolated, cultured, and then seeded onto the inner surface of bovine jugular vein patches at a density of 1.5 x 10(5) per cm2. Specimens were cultured for five days and then examined using scanning electron microscopy (SEM) and immunohistochemical staining for von Willebrand factor. RESULTS: SEM showed confluent lining of cultured ovine jugular vein endothelial cells similar to native endothelium. Von Willebrand factor staining for cells on the surface of patches was positive. CONCLUSION: The photooxidatively stabilized bovine jugular vein patches were not cytotoxic, and their in-vitro endothelialization was possible. PMID- 14565717 TI - Clinical experience and Doppler echocardiographic assessment of the first one hundred ATS AP (advanced performance) prosthetic valve in the aortic position. AB - BACKGROUND AND AIM OF THE STUDY: The ATS Medical mechanical bileaflet valve is composed of pyrolitic carbon and is available in two forms: Standard and Advanced Performance. The study aim was to evaluate the clinical results and define, in blinded manner, the Doppler echocardiographic characteristics of normally functioning ATS AP prostheses with respect to their size. METHODS: One hundred patients (63 men, 37 women; mean age 63.6 +/- 10.6 years) were studied between January 1996 and February 1999. Doppler echocardiography was performed at least three months after valve replacement (mean 1.68 +/- 0.86 months; range: 3 months to 3 years). RESULTS: The in-hospital mortality was 3%, and there were four late deaths. None of the deaths was valve-related. Thromboembolic and anticoagulant related hemorrhagic rates were 0.55% per patient-year (pt-yr) and 1.1% per pt-yr, respectively. Maximum and mean gradients were calculated using the simplified Bernoulli equation. Functional valve surface area was assessed using the continuity equation and time-velocity integrals using echographic measurements to calculate the subaortic surface. The Doppler velocity index was obtained from the ratio of subaortic and transaortic velocities. For the most frequently used aortic valve (22 mm), the maximum pressure gradient was 18.67 +/- 8.31 mmHg, the mean gradient 9.97 +/- 3.84 mmHg, functional surface area 1.50 +/- 0.35 cm2, and Doppler velocity index 0.41 +/- 0.08. CONCLUSION: Based on Doppler echocardiographic characteristics, the new ATS Medical AP prosthesis, when implanted in the aortic position, has an excellent hemodynamic profile which compares favorably with that of similarly designed prostheses. The clinical results show a very low rate of thromboembolic and anticoagulant-related hemorrhagic events, perhaps due to the new design of the pivoting area. PMID- 14565718 TI - Thrombogenicity and reoperation of the St. Jude Medical Silzone valve: a comparison with the conventional St. Jude Medical valve. AB - BACKGROUND AND AIM OF THE STUDY: The St. Jude Medical Silzone valve was withdrawn from the market after an interim analysis of the AVERT study revealed a 2.76% incidence of paravalvar leakage leading to valve re-replacement, compared with only 1.02% in the control group. Additionally, an increased risk for thromboembolic complications was reported in different series of patients. METHODS: To monitor the consistency of these reports, the incidence of thromboembolic complications and paravalvar leakage was investigated in 113 patients who received a Silzone valve at the authors' institution, compared with 101 patients who, immediately prior to the Silzone group, received a conventional valve. RESULTS: No difference was found in the incidence of either thromboembolism or paravalvar leakage after either Silzone or conventional valve replacement. CONCLUSION: Differences between the AVERT study and the present study might reflect differences in operative technique or in patient groups, but this as yet is unknown. To clarify the prospects of the estimated 36,000 patients with Silzone valves world wide, further research should aim at identifying possible unique features of patient groups with a higher risk of paravalvar leakage leading to valve explantation. In brief, an increased rate of paravalvar leakage and/or thromboembolism does not apply to either valve replacement groups. PMID- 14565719 TI - Total correction of complete atrioventricular septal defect with tetralogy of Fallot. AB - BACKGROUND AND AIM OF THE STUDY: The authors' experience in surgical management of tetralogy of Fallot in association with complete atrioventricular septal defect (CAVSD-TOF) is reported. METHODS: Between January 1990 and September 2001, 146 children with CAVSD underwent complete correction. ASD closure was performed using a one-patch technique, with autologous pericardial patch. Seventeen patients presented with CAVSD-TOF, of whom nine (53%) had undergone previous palliation. Mean age at repair was 2.9 +/- 1.9 years. The mean gradient across the right ventricular outflow tract (RVOT) was 63 +/- 16 mmHg. Six patients (35%) required a transannular patch. RESULTS: Overall in-hospital mortality was 10%. Among patients with CAVSD-TOF, three died in hospital (18%); causes of death were progressive heart failure (n = 2) and multiple organ failure (n = 1). Two patients required mediastinal exploration due to significant bleeding, and two late reoperation for severe left atrioventricular valve regurgitation after intracardiac repair. Mean follow up was 36 +/- 34 months. All patients survived and are currently in NYHA class I or II. At follow up, mean gradient across the RVOT was 17 +/- 6 mmHg, significantly less than the preoperative value (p < 0.001). CONCLUSION: Complete repair in patients with CAVSD-TOF offers acceptable early and mid-term outcome in terms of mortality, morbidity and reoperation rate. Palliation prior to repair may be preferred in cases with small pulmonary arteries, or in severely cyanotic neonates. The RVOT should be managed as for isolated TOF, but a transatrial-transpulmonary approach is the surgery of choice. PMID- 14565721 TI - Isolated native tricuspid valve Candida endocarditis in a non-drug-addicted patient: case report and review of the literature. AB - A case is reported of isolated native tricuspid calve Candida parapsilosis endocarditis (INTVCE) in a male patient with no history of drug abuse or heart disease. The patient had received hyperalimentation and antibiotics for four months via a central venous catheter after abdominal surgery. He underwent successful treatment with tricuspid valve debridement, liposomal amphotericin (AmBisome) and fluconazole, and remained without relapse during an eight-year follow up. A literature review of 12 similar cases (including the present patient) without history of drug abuse or heart disease, dating from 1970, is included. PMID- 14565720 TI - Severe mitral valve involvement in a child with hypereosinophilia secondary to parasitic infection. AB - An 11-year-old boy with severe mitral regurgitation due to hypereosinophilia caused by infection with a filaria (Mansonella perstans) required mitral valve replacement with a prosthetic valve. During recurrent postoperative hypereosinophilia, the patient experienced severe mitral stenosis due to thrombosis of the mitral prosthesis. Despite adequate anticoagulation, the prosthesis had to be replaced as an emergency with a second prosthetic valve. Permanent control of the eosinophil count was achieved with chronic oral steroid administration. In contrast to other microfilariae, M. perstans is non-pathogenic to humans; nevertheless, longstanding hypereosinophilia may lead to severe cardiac involvement endangering the patient's life. PMID- 14565722 TI - Mitral homograft in the tricuspid position, aortic homograft and mitral plasty in a drug addict with multiple valve endocarditis. AB - The case is reported of multiple valve surgery using as little prosthetic material as possible in a drug addict with recurrent right and left bacterial endocarditis. The patient underwent aortic valve replacement with a cryopreserved aortic homograft, mitral repair and tricuspid valve replacement with a mitral homograft, using a modified technique. The indications and surgical options for tricuspid valve endocarditis in this patient group are discussed, with particular focus on technical aspects of using mitral homografts in the tricuspid position. PMID- 14565723 TI - Hypertrophic non-obstructive cardiomyopathy with aortic and mitral regurgitation: an unusual presentation for surgical option. AB - The case is described of a pediatric patient with hypertrophic non-obstructive cardiomyopathy (HNCM) presenting with aortic and mitral regurgitation. This 17 year-old boy had been followed for HNCM since he was aged 4 years. Serial echocardiography had been showing the emergence and progression of aortic regurgitation and mitral regurgitation since the patient was aged 6 years. He started to have syncope attacks at 16 years of age and fell into congestive heart failure at 17 years. He underwent successful mitral valve repair by means of double-orifice technique, and aortic valve repair with raphe triangular resection, cusp plication and subvalvular annuloplasty. PMID- 14565724 TI - Comment on derivation of sample size requirements for evaluating heart valves with constant risk events. PMID- 14565725 TI - On the distribution of potassium iodide to members of the public in anticipation of an accidental release of radioiodine. PMID- 14565726 TI - History of the solar particle event radiation doses on-board aeroplanes using a semi-empirical model and Concorde measurements. AB - Measurements during solar particle events with dosemeters flying permanently on board Concorde are used to develop a semi-empirical model, called SiGLE. The model is intended to calculate, for a given flight plan, the dose equivalent received during a solar particle event observed with ground-based neutron monitors. It is successfully in operation in the SIEVERT computerised system intended to improve monitoring of radiation dose received by aircrews, in application to a European Directive. The semi-empirical model is applied to evaluate, for most exposed routes, the radiation doses corresponding to the GLEs observed since 1942 with ion chambers or neutron monitors. The results for the largest GLEs observed in the past are discussed in terms of radiation risk, and guidelines are suggested concerning possible alerts to the aeroplanes in case of events of exceptional magnitude. PMID- 14565727 TI - On the conversion coefficients for cosmic ray dosimetry. AB - Calculations of fluence-to-effective dose conversion coefficients have typically been limited to the standard irradiation geometries of the human body: anterior to-posterior (AP), posterior-to-anterior (PA), lateral from the right side to the left side (RLAT), lateral from the left side to the right side (LLAT), rotational around the vertical axis (ROT), and isotropic incidence from all directions (ISO). In order to estimate the doses to air crew members exposed to cosmic radiation, the geometrical conditions of irradiation are usually assumed to be isotropic. However, the assumption of isotropic irradiation is in many cases invalid for the high energy component of the radiation field, which is often peaked in the forward direction. Therefore, it was considered useful to extend the calculations of conversion coefficients to other geometries. New sets of conversion coefficients fluence-to-effective dose are presented for the semi isotropic irradiation of the human body and for the irradiation from the top. Their application to cosmic ray dosimetry is discussed. PMID- 14565729 TI - Plutonium content of human placental tissues after occupational exposure. AB - The placenta and umbilical cord were obtained following a normal live delivery from a volunteer donor who had received an accidental inhalation intake of plutonium 12 years prior to her pregnancy (Case 0777). Her employer estimated the intake to be about 73 Bq Class W plutonium. Based on bioassay results and clearance models in use at that time, they calculated her body content at the beginning of pregnancy to be about 5.6 Bq with an average concentration of approximately 60 mBq kg(-1). The placenta and cord from this pregnancy, along with the placenta and cord from a donor with no known exposure to plutonium (Case 0835), were divided and assayed for plutonium by ultrasensitive fission track analysis at two collaborating laboratories. Placental 239Pu concentration values obtained by the two laboratories for Case 0777 agreed within a factor of 2 and were several-fold greater than for the control, Case 0835, as well as values that had been reported by others for unexposed populations. There was no elevated concentration of plutonium in the umbilical cord from the exposed person. The data yielded values of 0.16 and 0.27 for placental to maternal concentrations (CPl: CM) that were of the same order of magnitude as the value of 0.1 the ICRP calculated for intakes before pregnancy. PMID- 14565728 TI - UK laboratory intercomparison on internal dosimetry. AB - A laboratory intercomparison for internal dose assessment from a variety of intake scenarios is described. This is the first UK intercomparison using the revised ICRP Human Respiratory Tract and biokinetic models. Four United Kingdom laboratories participated and six cases were assessed. Overall, the agreement in internal dose assessments between laboratories was considered satisfactory with 79% of the assessed committed effective doses, e(50), for cases within a band of +/- 40% of the median value. The range (highest/lowest) in e(50) estimated by the laboratories was smallest (1.2) for a case involving inhalation of 137Cs. The range was greatest (6.0) for a case involving a wound with, and possible inhalation of, 238Pu, 239Pu and 241Am; the variation between laboratories in assessment of intakes could not be considered to be satisfactory in this case. Judgements on the most appropriate data to use in estimating intakes, choice of parameter values for use with the ICRP models and allowing for the effects of treatment with DTPA were important sources of variability between laboratories. PMID- 14565730 TI - Assessment of the doses received by the Cuban population from 40K contained in the body: modelling based on a neural network. AB - Potassium-40 constitutes the main natural source of potassium present in the body, which influences the effective dose received by people. With the aim of assessing the contribution of this component to the doses received by the Cuban population, a study intended to assess the doses was developed. For this purpose, a representative sample of the Cuban population was selected according to age and sex. The measurements were made using the whole-body counter (WBC) of the Center for Radiation Protection and Hygiene (CPHR). For dose estimations, a uniform distribution of potassium for the whole body was assumed. The methodology used was the one recommended by the ICRP. The values of annual effective dose range between 93 and 209 microSv for females and between 123 and 212 microSv for males. The annual average effective dose for members of the public was estimated as 150 +/- 40 microSv, taking into account the experimental data and the specific features of the Cuban population. With the dose values obtained, it was possible to model dose estimates by means of a neural network, which was trained with the results obtained and using as starting data the sex, age, height and corporal weight of people studied. The modelobtained allows consideration of the anatomical features of each person in the estimation of doses. PMID- 14565731 TI - Seasonal correction factors for estimating radon exposure in dwellings in France. AB - Indoor radon concentrations are subject to seasonal variation with a maximum in winter and a minimum in summer. Procedures to correct for seasonal variation are necessary in order to get an unbiased estimate of the annual average radon concentration from data based on short-period radon measurements. To obtain correction factors, we apply the model developed by Pinel et al to the French database of indoor radon measurements (measurements performed as part of the indoor radon case-control study and of the national radon measurement campaign). For 6-month measurements, the correction factors vary from 0.87 to 1.17 and agree with those previously published. These results might be applicable when assessing indoor radon concentrations with regard to recommended action levels. PMID- 14565732 TI - The weekly measurement deviations of indoor radon concentration from the annual arithmetic mean. AB - The difference between weekly measurements and the annual arithmetic mean of radon indoor concentration CRn,Indoor was studied in the Czech Republic. The deviations were analysed for 1537 weekly measurements which were consecutively obtained in 29 rooms over a period of 1 year and the annual arithmetic mean was calculated for each particular room. The relationship of the deviations to three meteorological parameters (i.e. outside temperature, atmospheric pressure, and weekly rainfall) and to the sequential number of a calendar week was studied. The effect of atmospheric pressure and weekly rainfall was not significant. The deviation between a weekly measurement and the annual arithmetic mean depended significantly on outside weekly average temperatures. If the average outside weekly temperature was below 10 degrees C, the radon concentration was systematically higher than that of the annual arithmetic mean. The deviation variability was lower up to a temperature of 10 degrees C. If the weekly average outdoor temperature was higher than 10 degrees C, the uncertainty of a weekly measurement of radon concentration was also higher. PMID- 14565733 TI - Body potassium content and 40K radiation dose to Iranian subjects. AB - In this study 215 Iranian subjects including 180 males and 35 females were investigated for potassium body contents, its variations with age, weight, body build index, and slenderness, as well as the radiation dose due to 40K. The average total body potassium is 147 +/- 22 g for males and 100 +/- 16 g for females. The average potassium concentrations for males and females are 1.98 +/- 0.34 and 1.60 +/- 0.29 g kg(-1) body weight, respectively. The potassium content decreases with age for both sexes. The 40K concentration decreases with slenderness in both sexes, particularly in females. The total body potassium increases with body build index for males, while its value is constant for females less than 40 y, and increases for females more than 40 y. The average effective doses due to 40K are 187 +/- 32 microSv y(-1) for males and 150 +/- 27 microSv y(-1) for females. PMID- 14565734 TI - On the advantages and disadvantages of optically stimulated luminescence dosimetry and thermoluminescence dosimetry. AB - The use of thermoluminescence as a method for the dosimetry of ionising radiation has been established for many decades and has been unquestionably successful. It is therefore difficult to imagine how any new technique could easily supplant it. Perhaps optically stimulated luminescence dosimetry should not be characterised as an entirely new technique, but rather a development of the well established technology that may be considered superior in some respects. As is obvious from the tenor of this debate, our two participants are longtime colleagues who have had numerous discussions on this topic. No doubt, many of us will have similar discussions in our own institutions as we weigh the advantages and disadvantages of TLD versus OSL. PMID- 14565735 TI - "Thinking upstream" to confront prematurity. PMID- 14565737 TI - Caring for women living with Sjogren's syndrome. AB - T his article advises nurses about providing care to women who are living with Sjogren's syndrome. A chronic, autoimmune condition, Sjogren's syndrome affects mostly women and leads to visual, vocal, and vaginal problems related to dryness. A review of the literature indicates that not many women are aware of Sjogren's, and when diagnosed, are not informed about the possibility that they will have the vaginal dryness associated with the condition. Nursing implications include interventions that involve pharmacologic and nonpharmacologic methods to moisturize women's eyes, mouth, and vagina, increasing the comfort of those with Sjogren's syndrome. PMID- 14565736 TI - Safe criteria and procedure for kangaroo care with intubated preterm infants. AB - Kangaroo care (KC) was safely conducted with mechanically ventilated infants who weighed less than 600 grams and were less than 26 weeks gestation at birth. These infants, ventilated for at least 24 hours at the time of the first KC session, were considered stable on the ventilator at low settings (intermittent mandatory ventilation < 35 breaths per minute and FiO2 < 50%), had stable vital signs, and were not on vasopressors. A protocol for implementation of KC with ventilated infants that uses a standing transfer, with two staff members assisting to minimize the possibility of extubation, is presented. Also discussed is the positioning of the ventilator tubing during KC. This protocol was implemented without any accidental extubation throughout an experimental research study. The criteria and protocol were compared to those available in published reports and revealed many similar elements, providing additional support for the recommended protocol. No adverse events occurred with the criteria and protocol reported here, suggesting that they can be adopted for broader use. PMID- 14565738 TI - A critical pathway for intimate partner violence across the continuum of care. AB - OBJECTIVE: The authors developed an interdisciplinary critical pathway for intimate partner violence (IPV) assessment and intervention for use across health care settings. Intimate partner violence may be emotional, physical, and/or sexual and involves coercion and control by one partner over the other. DESIGN: A pathway developed with input from focus groups of battered women was subjected to a modified Delphi technique to improve the pathway's scientific accuracy and feasibility. SETTING: The study was conducted in one urban, one suburban, and one rural hospital with IPV advocacy programs in the mid-Atlantic region of the United States. PATIENTS/PARTICIPANTS: Four researchers and 13 clinicians participated in the validation, with at least one physician, one nurse, and one social worker or IPV counselor from each hospital. MAIN OUTCOME MEASURES: Each element of the pathway was analyzed for the degree of consensus on scientific accuracy and feasibility. RESULTS: Consensus on the scientific accuracy and feasibility of the pathway was achieved after three rounds of the Delphi process. CONCLUSIONS: This is the first known critical pathway for IPV. It addresses physical and mental health and safety and has content validity affirmed by an interdisciplinary panel of experts. Further process and outcome evaluation is warranted and invited. PMID- 14565739 TI - Mother-newborn contact in a randomized trial of kangaroo (skin-to-skin) care. AB - OBJECTIVE: To describe the type and percent time of contact 0-48 hours postbirth for mother-preterm newborn (infant) dyads given kangaroo care (skin-to-skin) or standard care (controls). DESIGN: Randomized controlled trial with assignment by computerized minimization to kangaroo care (n = 48) or control (n = 43). SETTING: Postpartum units and neonatal intensive-care units (NICU). PARTICIPANTS: Preterm infants 32 to less than 37 weeks gestation and their mothers. INTERVENTION: Kangaroo (skin-to-skin, SS) care (KC). MAIN OUTCOME MEASURES: Type and percent time of mother-infant contact (SS versus holding wrapped in blankets). RESULTS: Analyses were based on four groups: assignment for infants in each group to postpartum or NICU. For KC dyads, SS postpartum was 22.0%; SS NICU was 7.5%. KC wrapped holding postpartum was 11.6%; NICU was 1.8%. For control dyads, wrapped holding postpartum was 13.9%; NICU was 6.1%. CONCLUSION: Amount of SS was much less than expected. Reasons include unavailability of infants or mothers and hospital staff interrupting contact. However, KC postpartum dyads were held wrapped almost as often as control postpartum dyads. Total contact time for KC dyads (SS plus wrapped) was more than double that of controls. These data suggest that hospital and social supports for families are needed to facilitate early initiation of SS, prolonged periods of mother-infant SS contact, and reduction of maternal stress. PMID- 14565740 TI - Situational factors involved in college students' safer and risky sexual encounters. AB - OBJECTIVE: To ascertain the differences between the situational factors of risky and safer sexual encounters in college students. DESIGN: Descriptive. SETTING: Community-dwelling college students from a large Midwestern university. PARTICIPANTS: Participants were 84 college students ages 18 to 20 years. Fifty five were female (67.5%), the majority (95.2%) were White, and all reported being heterosexual. MAIN OUTCOME MEASURE: Use of safer or risky practices in last or most memorable sexual encounter. RESULTS: Important situational characteristics of safer encounters were negative ambiance of the place, type of partner, unattractive qualities of the partner, communication with the partner, and having the encounter because of feeling desire. Important situational characteristics of risky encounters were a cozy ambiance, partner's attractiveness inducing desire, asking directly for the encounter, and encounters occurring after celebration or partying. CONCLUSIONS: Many characteristics were similar for both risk groups. However, practitioners can tailor interventions to make clients aware of factors that were more prevalent in risky sexual encounters, such as having the encounter with a boyfriend or girlfriend and having an encounter after drinking alcohol and partying. PMID- 14565742 TI - Resources for evidence-based practice, September-October 2003. PMID- 14565741 TI - The influence of perinatal loss on anxiety in multigravidas. AB - OBJECTIVE: To compare multigravid women with and without a history of perinatal loss on state anxiety, pregnancy anxiety, and optimism. DESIGN: Comparative descriptive; cross-sectional. SETTING: Private obstetric offices in a small northeastern city in the United States. PARTICIPANTS: The sample included 160 women who were between 17 and 28 weeks gestation: 96 multigravidas with no history of loss and 74 women with a history of one or two losses. MAIN OUTCOME MEASURES: State anxiety, pregnancy anxiety, optimism, and perinatal loss history. RESULTS: No group differences were found on demographic variables, state anxiety, or optimism. However, pregnancy anxiety was higher in women with a history of perinatal loss. Pregnancy anxiety was also correlated with desire to see care provider more often and number of phone calls between visits, and was not correlated with the number of living children. CONCLUSION: Women experiencing pregnancy subsequent to perinatal loss have greater pregnancy anxiety: That is, they are more concerned about their pregnancies and their babies than women without a history of perinatal loss. State anxiety and optimism do not differentiate these two groups. This heightened anxiety should be acknowledged and more frequent contact with the care provider should be offered. PMID- 14565743 TI - Preterm birth. PMID- 14565744 TI - Preterm labor and birth: what have we learned in the past two decades? AB - Preterm births in the United States reached a 20-year high of 11.9% in 2001. Preterm and low-birth-weight births are the end result of multiple pathways. This article examines two decades of multidisciplinary research related to preterm birth from both individual and ecologic perspectives. The difficulties in identifying women who will have preterm birth, risk factors amenable to change in the preconception and prenatal periods, and strategies for intervention are described, along with maternal treatment to improve infant outcomes. Future directions for nursing practice and research are suggested. PMID- 14565745 TI - Psychosocial pathways to prematurity: changing our thinking toward a lifecourse and community approach. AB - The purpose of this article is to explore the psychosocial antecedents of prematurity. Emphasis is on conceptual areas and supporting literature for (a) the contexts in which prematurity occurs and the diversity of women's experiences; (b) a lifecourse approach to prematurity that highlights allostatic load and the accumulation of trauma and loss in possible prematurity pathways; and (c) diverse psychosocial/biological pathways and mechanisms of prematurity processes. Pathways examining psychosocial and prematurity connections will be explicated, including antecedents and outcomes other than stress proneness and vulnerability. Implications for research are logically derived from a focus on the impact of social context on individual outcomes through multilevel models and methods. Clinical implications are derived from the social contexts, lifecourse, and multiple pathways focus of the article and include increasing social cohesion in communities, population health strategies, particular psychosocial interventions, and attentive listening. PMID- 14565746 TI - Nursing's contribution to the literature on preterm labor and birth. AB - Although preterm labor and birth continues to plague us as an unsolved mystery in maternal/child health, much has actually been learned over the past few decades. Most reviews of the preterm labor literature examine the research in an interdisciplinary manner; this article instead looks at the active role that nurses have taken in the research concerning preterm labor and birth. This article will categorize research conducted by nurses over the past 30 years on preterm labor and birth, emphasizing some of the changes in nursing practice engendered by nursing's important research effort. PMID- 14565747 TI - Psychological and physiological stress: impact on preterm birth. AB - Stress increases corticotropin-releasing hormone and may ultimately result in increased uterine contractility. Stress also increases cytokine production, which independently may lead to preterm birth or increase susceptibility to infection, thereby increasing the risk of preterm birth. Finally, stress may change health behaviors that lead to preterm birth. Research findings on the relationship between stress and preterm birth have been contradictory. In this article, the authors propose a model of the relationship between stress and preterm birth, evaluate the research on stress and pregnancy outcomes, and discuss the implications for nursing practice and research. PMID- 14565749 TI - State of the science: achievements and challenges across the spectrum of care for preterm infants. AB - The spectrum of care for preterm infants includes the perinatal and immediate neonatal periods, the initial hospitalization period including neonatal intensive care, transition to discharge, and from discharge through the first year of life. Care issues are sometimes lifelong. Advances and achievements of the past 20 years, particularly during the perinatal period and in neonatal intensive care, have resulted in significant increases in survivability of even the smallest and least mature infants. Challenges remain, particularly in establishing evidence based standards of nursing practice in areas such as transition to oral feedings, breastfeeding in the intensive-care nursery, and developmentally based care, and in reducing short- and long-term morbidities in children born prematurely. This article illustrates achievements and challenges across the spectrum of care for preterm infants in the 1st year of life. PMID- 14565748 TI - Implementing evidence-based practice: reducing risk for low birth weight through pregnancy smoking cessation. AB - In 1989, the Association of Women's Health, Obstetric and Neonatal Nurses (AWHONN) developed a research utilization program to integrate evidence into practice areas where there were large discrepancies between research evidence and clinical practice. The current program, renamed Research-Based Practice (RBP), uses translational research methods to build from evidence such as that in the Cochrane database and to create protocols for integration of research directly into clinical practice. This article describes the development of the sixth project (RBP6), in which an evidence-based protocol to address smoking in pregnancy was integrated into clinical practice. The protocol includes screening women using descriptive statements and integrates the 5 As (ask, advise, assess, assist, arrange) into prenatal and postpartum care at every visit. By integrating smoking cessation counseling into care, nurses may reduce the risk of low birth weight among pregnant women in both the United States and Canada. PMID- 14565750 TI - Botulinum neurotoxin type A. PMID- 14565751 TI - New-generation implant arthroplasties of the finger joints. AB - Prosthetic replacement in the hand must address such unique challenges as preservation of the collateral ligaments, tendon balancing, and stability. Some recently developed implant arthroplasties of the metacarpophalangeal and proximal interphalangeal joints have anatomically designed articular components; others have non-cemented, press-fit, carefully contoured intramedullary stems. The rationale behind developing the unlinked or semiconstrained prosthesis with anatomic geometry is that it would create balanced forces across the joint. Low profile, anatomically designed implants limit the amount of bone removed and preserve the integrity of the collateral ligaments. A metacarpophalangeal joint implant with an elliptical metacarpal head and a nonfixed center of rotation can enhance stability in flexion through greater articular contact. A proximal interphalangeal joint implant that preserves the collateral ligaments also can achieve improved stability. Component loosening is not an early complication with these recent designs, and arc of motion is satisfactory. PMID- 14565752 TI - Lower extremity angular malunion: evaluation and surgical correction. AB - The lower extremity has a mechanical axis with joint orientation that allows joint longevity and efficiency in bipedal gait. When normal alignment is lost because of trauma or other conditions, deviations from this anatomic norm may be deleterious to long-term joint function. In fractures that have healed with angular malunion, all facets of the deformity must be carefully considered, including alteration in length, rotation, alignment, and translation. Once all elements are fully defined, the effects of the malunion on mechanical axis and joint orientation can be understood. Techniques for surgical correction include wedge, dome, and oblique osteotomies and distraction osteogenesis. Each method possesses characteristics appropriate for certain clinical situations. Judicious patient selection and thoughtful preoperative planning may allow restoration of normal mechanics. PMID- 14565753 TI - Common rotational variations in children. AB - Most rotational variations in young children, such as in-toeing, out-toeing, and torticollis, are benign and resolve spontaneously. Understanding the normal variations in otherwise healthy children is vital to identifying true structural abnormalities that require intervention. A deliberate assessment of the rotational profile is necessary when evaluating children who in-toe or out-toe. In-toeing is usually attributable to metatarsus adductus in the infant, internal tibial torsion in the toddler, and femoral anteversion in children younger than 10 years. Out-toeing patterns largely result from external rotation hip contracture, external tibial torsion, and external femoral torsion. Although congenital muscular torticollis is the most common explanation for the atypical head posture in children, more serious disorders, including osseous malformations, inflammation, and neurogenic disorders, should be excluded. PMID- 14565754 TI - Chronic massive rotator cuff tears: evaluation and management. AB - Most studies of rotator cuff repairs report high success rates. However, the majority of these studies combine the results of surgical management of rotator cuff tears of various sizes; few published reports specifically evaluate the management of chronic massive tears. Chronic massive rotator cuff tears may be acute traumatic, chronic atraumatic, or acute-on-chronic. A detailed history and thorough physical examination often are sufficient to establish the diagnosis. Radiographic evaluation can reveal osseous changes suggestive of pathology. Magnetic resonance imaging can determine the size of rotator cuff tears and status of the muscles but generally is not necessary for patients who are not candidates for surgery. Chronic massive rotator cuff tears without glenohumeral arthritis can be managed nonsurgically or with sub-acromial debridement, rotator cuff repair, or rotator cuff reconstruction. However, treatment of these patients is challenging, and results are comparatively inferior to those of treating patients with smaller rotator cuff tears. PMID- 14565755 TI - Pediatric soft-tissue tumors. AB - Soft-tissue tumors in children (<18 years) are a heterogeneous group of lesions. Masses may be asymptomatic or associated with pain or discomfort. Although most lesions are benign, developing an appropriate differential diagnosis requires knowledge of the clinical and radiographic characteristics of tumors and tumorlike conditions in children. A thorough history and physical examination, followed by appropriate imaging studies, when indicated, can establish a correct diagnosis and help determine appropriate treatment recommendations. PMID- 14565756 TI - Use of physical forces in bone healing. AB - During the past two decades, a number of physical modalities have been approved for the management of nonunions and delayed unions. Implantable direct current stimulation is effective in managing established nonunions of the extremities and as an adjuvant in achieving spinal fusion. Pulsed electromagnetic fields and capacitive coupling induce fields through the soft tissue, resulting in low magnitude voltage and currents at the fracture site. Pulsed electromagnetic fields may be as effective as surgery in managing extremity nonunions. Capacitive coupling appears to be effective both in extremity nonunions and lumbar fusions. Low-intensity ultrasound has been used to speed normal fracture healing and manage delayed unions. It has recently been approved for the management of nonunions. Despite the different mechanisms for stimulating bone healing, all signals result in increased intracellular calcium, thereby leading to bone formation. PMID- 14565757 TI - Thoracolumbar fracture management: anterior approach. AB - The surgeon who treats patients with spine trauma must be able to apply a variety of management techniques to achieve optimal care of the patient. The anterior surgical approach is appropriate for some thoracolumbar burst fractures in patients with neurologic deficit and without posterior ligamentous injury. Surgery is most often indicated for patients with incomplete deficit, especially those with a large retropulsed fragment, marked canal compromise, severe anterior comminution, or kyphosis <30 degrees. This approach provides excellent visualization of the anterior aspect of the dura mater for decompression. Reconstruction of the anterior body defect can be done with autograft, allograft, or a cage. Supplementation of the graft with anterior internal fixation helps prevent kyphosis. Clinical results demonstrate improved neurologic function in most patients as well as low pseudarthrosis rates. In patients with incomplete deficit, improvement in neurologic function usually can be expected with few complications. PMID- 14565758 TI - The unstable patella after total knee arthroplasty: etiology, prevention, and management. AB - In total knee arthroplasty, most complications related to the extensor mechanism are caused by patellar maltracking or instability. Patellar maltracking may result from component malpositioning and limb malalignment, prosthetic design, improper patellar preparation, or soft-tissue imbalance. Patellofemoral instability likely results most frequently from internal malrotation of the femoral or tibial components. Although a patellofemoral radiograph may display the lateral subluxation of the patella, only computed tomography can quantify rotational malalignment of the femoral or tibial component. Nonsurgical treatment is generally unsuccessful; major malposition of components is best managed by implant revision. In the absence of component malposition, proximal realignments (lateral patellar retinacular release with lateral advancement of the vastus medialis obliquus muscle) or tibial tubercle transfers have been used. Surgical procedures on the patellar tendon itself may risk rupture of the extensor mechanism. PMID- 14565759 TI - Protein-protein interactions involving DNA glycosylases. PMID- 14565760 TI - Reactivity with Tris(hydroxymethyl)aminomethane confounds immunodetection of acrolein-adducted proteins. AB - The toxic alpha,beta-unsaturated aldehyde acrolein readily attacks proteins, generating adducts at cysteine, histidine, and lysine residues. In this study, rabbit antiserum was raised against acrolein-modified keyhole limpet hemocyanin in the expectation that it would allow immunodetection of adducted proteins in biological samples. Using slot-blot and enzyme-linked immunosorbent assays, the antiserum detected acrolein-modified protein with high sensitivity and specificity. Adduct immunodetection was strongly inhibited by acrolein-modified polylysine but not polyhistidine. Efforts to develop a Western blotting method for detecting adducted proteins in cell lysates were hampered by irreproducible outcomes, evidently due to adduct instability during SDS-PAGE. Indeed, adducts generated via brief exposure of a model protein to acrolein displayed pH- and concentration-dependent instability to tris(hydroxymethyl)aminomethane (Tris), a nucleophilic buffer used in protein electrophoresis. The effect was most striking when Tris solutions were buffered to pH 8.0 and higher. In contrast, adducts formed during extended exposure to acrolein (> or =60 min) were completely stable to Tris. The time dependence of susceptibility raised the possibility that Tris interfered with specific steps in lysine modification, which involves stepwise Michael addition of two molecules of acrolein to the same residue, followed by condensation and dehydration to form a heterocyclic adduct, N(epsilon)-(3-formyl 3,4-dehydropiperidino)lysine. We hypothesize that carbonyl-retaining Michael adducts may react with Tris by forming imines with the primary amine of the buffer. Consistent with this idea, triethanolamine, a tertiary amine buffer unable to form imines, had no effect on acrolein-adducted protein. These effects of Tris may explain difficulties in the detection of acrolein-adducted proteins during conventional Western blotting procedures. PMID- 14565762 TI - 1H NMR pattern recognition and 31P NMR studies with d-Serine in rat urine and kidney, time- and dose-related metabolic effects. AB - Proton NMR spectroscopy of urine has previously been used to gain insight into the site and mechanism of toxic injury to the kidney. d-Serine injures the rat kidney, causing selective necrosis of the proximal straight tubules. Damage is accompanied by proteinuria, glucosuria, and amino aciduria, the latter preceding the onset of necrosis. This study has employed (1)H NMR spectroscopy of urine and (1)H NMR and (31)P NMR spectroscopy of kidney extracts to examine the nephrotoxic action of d-serine. Urine was collected 0-8 h (all doses) and 8-24, 24-48, 48-72, 72-96, and 96-120 h (500 mg/kg only) postdosing from Alderley Park rats given d serine (62.5, 125, 250, and 500 mg/kg ip). (1)H NMR spectra were monitored for markers of tubular damage. Additionally, ATP and ADP were quantitated in kidney perchloric acid extracts, prepared after 0.5, 1, 2, 4, and 8 h (500 mg/kg) to assess energy status; serine was also measured in these samples. At 500 mg/kg, glucosuria, amino aciduria, and reduced citrate, alpha-ketoglutarate, and succinate were observed in urine at 0-8 h. Furthermore, serine and pyruvate levels were elevated at this time. After 8-24 h, similar changes were observed; however, they were more severe reflecting the development of the lesion prior to recovery. These perturbations were dose-related, in particular, for serine and pyruvate, with no alterations seen at 62.5 mg/kg. Kidney serine concentration rapidly increased, where it was maximal after 30 min and cleared by 8 h. A decline in ATP, to approximately 60-70% of control, was observed within the kidney at 2-4 h postdosing, when necrosis first becomes evident suggesting that mitochondrial function might be impaired in the early stages of d-serine-induced nephrotoxicity. The use of NMR spectroscopy has given a comprehensive overview of the effects of d-serine in vivo. Information on the excretion of serine and its effect on renal energy metabolism provides insight into the possible mechanism of renal tubule injury. PMID- 14565761 TI - Probing the possible molecular origin of the highly selective toxicity of antimalarial peroxide qinghaosu (artemisinin). AB - The radicals derived from qinghaosu (artemisinin) showed strong tendency for intramolecular reactions and thus are unlikely to cause so much damage to cells as simple alkyl radicals, giving a clue to the extraordinarily low toxicity of this radical precursor at therapeutic doses. PMID- 14565763 TI - Structural basis of binding and inhibition of novel tarantula toxins in mammalian voltage-dependent potassium channels. AB - Voltage-dependent potassium channel Kv2.1 is widely expressed in mammalian neurons and was suggested responsible for mediating the delayed rectifier (I(K)) currents. Further investigation of the central role of this channel requires the development of specific pharmacology, for instance, the utilization of spider venom toxins. Most of these toxins belong to the same structural family with a short peptide reticulated by disulfide bridges and share a similar mode of action. Hanatoxin 1 (HaTx1) from a Chilean tarantula was one of the earliest discussed tools regarding this and has been intensively applied to characterize the channel blocking not through the pore domain. Recently, more related novel toxins from African tarantulas such as heteroscordratoxins (HmTx) and stromatoxin 1 (ScTx1) were isolated and shown to act as gating modifiers such as HaTx on Kv2.1 channels with electrophysiological recordings. However, further interaction details are unavailable due to the lack of high-resolution structures of voltage sensing domains in such mammalian Kv channels. Therefore, in the present study, we explored structural observation via molecular docking simulation between toxins and Kv2.1 channels based upon the solution structures of HaTx1 and a theoretical basis of an individual S3(C) helical channel fragment in combination with homology modeling for other novel toxins. Our results provide precise chemical details for the interactions between these tarantula toxins and channel, reasonably correlating the previously reported pharmacological properties to the three-dimensional structural interpretation. In addition, it is suggested that certain subtle structural variations on the interaction surface of toxins may discriminate between the related toxins with different affinities for Kv channels. Evolutionary links between spider peptide toxins and a "voltage sensor paddles" mechanism most recently found in the crystal structure of an archaebacterial K(+) channel, KvAP, are also delineated in this paper. PMID- 14565764 TI - Computer-aided knowledge generation for understanding skin sensitization mechanisms: the TOPS-MODE approach. AB - The TOPS-MODE (topological substructural molecular descriptors) approach is used to derive models for understanding the molecular structural contribution to skin sensitization. A data set of 93 compounds was used in the development of the models; 29 new skin sensitization values (EC3) are reported here for the first time. The models developed possess high predictivity and have been validated through the use of cross-validation and external validation sets. The models have enabled the formulation of potential new structural alerts far faster and using less data than typically required by traditional approaches. Structural contributions to skin sensitization for various classes of chemicals are presented on the basis of bond contributions. The models have also been able to identify potential structural alerts for chemicals requiring metabolic activation. PMID- 14565765 TI - Structural differentiation of diastereomeric benzo[ghi]fluoranthene adducts of deoxyadenosine by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and postsource decay. AB - The product ion formation characteristics of four diastereomeric deoxyadenosine adducts formed by the reaction of the syn and anti diastereomers of trans-3,4 dihydroxy-5,5a-epoxy-3,4,5,5a-tetrahydrobenzo[ghi]fluoranthene are studied by matrix-assisted laser desorption ionization and postsource decay (PSD) to determine fragmentation pathways that may permit differentiation of their structures. The two adducts derived from each diol-epoxide with DNA differ in structure based on the cis/trans arrangement of the 3'-hydroxyl group on the benzo[ghi]fluoranthene (B[ghi]F) and the adenine base bound to the B[ghi]F 5a carbon. The two adduct diastereomers with the cis adenine-3'-hydroxyl configuration produce product ions at m/z 394 and m/z 510 formed by the loss of water that are not observed in the PSD spectra of the two trans isomers. The data suggest a mechanism of water loss that is initiated by a hydrogen-bonding interaction between the charge-bearing proton on the N1 atom and the 3'-hydroxyl oxygen on the polycyclic aromatic hydrocarbon (PAH). Fragmentation is initiated by the transfer of the adenine N1 proton from the nitrogen to the PAH 3'-hydroxyl oxygen and inductive cleavage of the C3-O(3) bond to form a benzylic carbocation on B[ghi]F. The proposed mechanism is supported by semiempirical molecular modeling calculations. PMID- 14565766 TI - Formation of vinylogous compounds in model Maillard reaction systems. AB - The thermal degradation over temperature and time of selected amino acids (Asp, Gln, and Glu) in the presence of reducing sugars was investigated in low moisture model systems. Copyrolysis of glucose-Asp mixtures led to the release of acrylic acid, attaining >5 mmol/mol Asp at 230 degrees C after 5 min. Spurious amounts of 3-butenamide were detected upon heating Gln together with a carbonyl source. Apparently, intramolecular cyclization is favored to procure 2-pyrrolidinone, reaching levels >3 mmol/mol above 230 degrees C. 2-Pyrrolidinone was also formed in comparable amounts in pyrolyzed sugar-Glu mixtures, indicating that the Maillard reaction may be an important contributor to the formation of 2 pyrrolidinone in certain cooked foods. The chemical route to acrylic acid and 3 butenamide is probably analogous to that described for acrylamide recently. Evidence is also presented that acrylic acid may be an intermediate in the formation of acrylamide, and yields could be augmented by coincubation of fructose-Asp with certain amino acids such as Gln, reaching approximately 5% of the yield obtained by the Asn route. A computational study to determine the reactivity of the vinylogous products indicated a reduced ability of 3-butenamide as compared to acrylamide to form stable intermediates by Michael nucleophilic addition. Acrylamide and acrylic acid exhibited a similar theoretical reactivity potential toward nucleophiles. No information is as yet available on the occurrence of acrylic acid in cooked foods. Extensive toxicological evaluation indicates that acrylic acid is of no concern at the amounts to be expected in foods. PMID- 14565767 TI - Identification of DNA adducts using HPLC/MS/MS following in vitro and in vivo experiments with arylamines and nitroarenes. AB - Arylamines and nitroarenes are suspected of playing a key role in chemical carcinogenesis. Therefore, the study of DNA adduct formation is an important step to determine the genotoxic potential of these compounds. Calf thymus DNA was modified in vitro by reaction with activated N-hydroxyarylamines: 2-chloroaniline (2CA), 4-chloroaniline (4CA), 2-methylaniline (2MA), 4-methylaniline (4MA), 2,4 dimethylaniline (24DMA), 2,6-dimethylaniline (26DMA), 2-aminobiphenyl (2ABP), 3 aminobiphenyl (3ABP), and 4-aminobiphenyl (4ABP). Female Wistar rats (n = 2) were given a single dose of the above arylamines and their analogous nitro derivatives by oral gavage and sacrificed after 24 h. Hepatic DNA and in vitro modified DNA were hydrolyzed enzymatically to individual 2'-deoxyribonucleosides. Adducts were determined using HPLC/MS/MS by comparison to synthesized standards. The hydrolysis efficiency was monitored by HPLC with UV detection. Each arylamine described above formed adducts to 2'-deoxyguanosine and 2'-deoxyadenosine after in vitro reaction with DNA. DNA adducts were found in rats dosed with 4ABP or with 4-nitrobiphenyl. DNA adducts were not detected in rats dosed with 2CA, 4CA, 2MA, 4MA, 24DMA, 26DMA, 2ABP, 3ABP, 2-chloronitrobenzene, 4-chloronitrobenzene, 2 nitrotoluene, and 4-nitrotoluene. All compounds formed hydrolyzable hemoglobin adducts. Therefore, biologically available N-hydroxyarylamines yielded hemoglobin adducts but not hepatic DNA adducts, except for 4ABP. PMID- 14565768 TI - Nitrosation, nitration, and autoxidation of the selective estrogen receptor modulator raloxifene by nitric oxide, peroxynitrite, and reactive nitrogen/oxygen species. AB - The regulation of estrogenic and antiestrogenic effects by selective estrogen receptor modulators (SERMs) provides the basis for use in long-term therapy in cancer chemoprevention and postmenopausal osteoporosis. However, the evidence for carcinogenic properties within this class requires study of potential pathways of toxicity. There is strong evidence for the elevation of cellular levels of NO in tissue treated with SERMs, including the benzothiophene derivative, raloxifene, in part via up-regulation of nitric oxide synthases. Therefore, the reactions of 17beta-estradiol (E(2)), raloxifene, and an isomer with NO, peroxynitrite, and reactive nitrogen/oxygen species (RNOS) generated from NO(2)(-)/H(2)O(2) systems were examined. Peroxynitrite from bolus injection or slow release from higher concentrations of 3-morpholinosydnonimine (SIN-1) reacted with the benzothiophenes and E(2) to give aromatic ring nitration, whereas peroxynitrite, produced from the slow decomposition of lower concentrations of SIN-1, was relatively unreactive toward E(2) and yielded oxidation and nitrosation products with raloxifene and its isomer. The oxidation and nitrosation products formed were characterized as a dimer and quinone oxime derivative. Interestingly, the reaction of the benzothiophenes with NO in aerobic solution efficiently generated the same oxidation products. Stable quinone oximes are not unprecedented but have not been previously reported as products of RNOS-mediated metabolism. The reaction of glutathione (GSH) with the quinone oxime gave both GSH adducts from Michael addition and reduction to the corresponding o-aminophenol. The ready autoxidation of raloxifene, observed in the presence of NO, is the first such observation on the reactivity of SERMs and is potentially a general phenomenon of significance to SERM chemical toxicology. PMID- 14565769 TI - Inhibition of peroxynitrite-mediated LDL oxidation by prenylated flavonoids: the alpha,beta-unsaturated keto functionality of 2'-hydroxychalcones as a novel antioxidant pharmacophore. AB - Prenylated 2'-hydroxychalcones and flavanones from the inflorescences of the female hop plant (Humulus lupulus) were shown to inhibit peroxynitrite-mediated oxidation of low-density lipoproteins (LDL) at low micromolar concentrations. LDL oxidation was induced by the peroxynitrite generator, 3-morpholinosydnonimine (SIN-1), and measured by the formation of conjugated dienes and thiobarbituric reactive substances. Human intake of prenylated chalcones and flavanones is mainly through beer, which contains up to 4 mg/L of these polyphenols. The two main oxidation products obtained by SIN-1 and peroxynitrite treatment of xanthohumol (XN), the principal prenylflavonoid of hops, were the aurone, auroxanthohumol (AUXN), and an endoperoxy derivative of XN, named endoperoxyxanthohumol (EPOX). In addition, the reaction produced smaller amounts of the nitro and nitroso derivatives of XN and EPOX. The formation of these nitrated products was enhanced in the presence of sodium bicarbonate (25 mM). SIN 1-induced formation of AUXN is considered to be a superoxide-mediated reaction, while the structure of EPOX points to a two electron oxidation reaction involving a Michael type addition with peroxynitrite as the nucleophile, followed by cyclization yielding a (1,2)-dioxepin-5-one ring structure. The flavanone isomer of XN, isoxanthohumol (IsoXN), unexpectedly showed a slight prooxidant effect instead of an inhibitory effect on LDL oxidation. Except for the formation of minor nitrated products, IsoXN remained largely unmodified upon treatment with SIN-1/peroxynitrite. Taken together, our results suggest that the alpha,beta unsaturated keto functionality of chalcones is most reactive toward superoxide and peroxynitrite anions. PMID- 14565770 TI - Detoxication pathways involving glutathione and epoxide hydrolase in the in vitro metabolism of chloroprene. AB - Chloroprene (2-chloro-1,3-butadiene, 1) is an important industrial chemical, which is carcinogenic in experimental animals and possibly in humans. It is metabolized to the monoepoxides, 2-chloro-2-ethenyloxirane (2a,b) and (1 chloroethenyl)oxirane (3a,b), together with electrophilic chlorinated aldehydes and ketones. This study has investigated the detoxication of these chloroprene metabolites in vitro by glutathione (GSH) and epoxide hydrolase (EH) in liver microsomes from Sprague-Dawley rats, B6C3F1 mice, and humans. In incubations of chloroprene with liver microsomes containing GSH, several GSH conjugates were identified. These were 1-hydroxy-4-(S-glutathionyl)butan-2-one (13), 1,4-bis-(S glutathionyl)butan-2-one (15), and (Z)-2-(S-glutathionyl)but-2-en-1-al (16). A fourth GSH conjugate was identified as either 2-chloro-3-hydroxy-4-(S glutathionyl)butene (12a,b) or 1-chloro-4-(S-glutathionyl)-butan-2-one (14), which were indistinguishable by LC/MS. Structural assignments of metabolites were based on chromatographic and spectroscopic comparisons with synthetic standards. There were significant differences between species in the amounts of 3a,b formed in microsomal incubations, the order being mouse > rat > human. Hydrolysis by microsomal EHs showed a distinct selectivity for S-(1-chloroethenyl)oxirane (3b) resulting in an accumulation of the R-enantiomer; the ratio of the amounts between species was 20:4:1 for mouse:rat:human, respectively. PMID- 14565771 TI - Preferential metabolic activation of N-nitrosopiperidine as compared to its structural homologue N-nitrosopyrrolidine by rat nasal mucosal microsomes. AB - N-Nitrosopiperidine (NPIP) is a potent rat nasal carcinogen whereas N nitrosopyrrolidine (NPYR), a hepatic carcinogen, is weakly carcinogenic in the nose. NPIP and NPYR may be causative agents in human cancer. P450-catalyzed alpha hydroxylation is the key activation pathway by which these nitrosamines elicit their carcinogenic effects. We hypothesize that the differences in NPIP and NPYR metabolic activation in the nasal cavity contribute to their differing carcinogenic activities. In this study, the kinetics of tritium-labeled NPIP or NPYR alpha-hydroxylation mediated by Sprague-Dawley rat nasal olfactory or respiratory microsomes were investigated. To compare alpha-hydroxylation rates of the two nitrosamines, tritiated 2-hydroxytetrahydro-2H-pyran and 2-hydroxy-5 methyltetrahydrofuran, the major NPIP alpha-hydroxylation products, and tritiated 2-hydroxytetrahydrofuran, the major NPYR alpha-hydroxylation product, were quantitated by HPLC with UV absorbance and radioflow detection. These microsomes catalyzed the alpha-hydroxylation of NPIP more efficiently than that of NPYR. K(M) values for NPIP were lower as compared to those for NPYR (13.9-34.7 vs 484 7660 muM). Furthermore, catalytic efficiencies (V(max)/K(M)) of NPIP were 20-37 fold higher than those of NPYR. Previous studies showed that P450 2A3, present in the rat nose, also exhibited this difference in catalytic efficiency. For both types of nasal microsomes, coumarin (100 muM), a P450 2A inhibitor, inhibited NPIP and NPYR alpha-hydroxylation from 63.8 to 98.5%. Furthermore, antibodies toward P450 2A6 inhibited nitrosamine alpha-hydroxylation in these microsomes from 68.8 to 78.4% whereas antibodies toward P450 2E1 did not inhibit these reactions. Further immunoinhibition studies suggest some role for P450 2G1 in NPIP metabolism by olfactory microsomes. In conclusion, olfactory and respiratory microsomes from rat nasal mucosa preferentially activate NPIP over NPYR with P450 2A3 likely playing a key role. These results are consistent with local metabolic activation of nitrosamines as a contributing factor in their tissue-specific carcinogenicity. PMID- 14565772 TI - Proteomic characterization of metabolites, protein adducts, and biliary proteins in rats exposed to 1,1-dichloroethylene or diclofenac. AB - A proteome profiling approach was used to compare effects of two toxicants, 1,1 dicloroethylene (DCE) and diclofenac, which covalently adduct hepatic proteins. Bile was examined as a potential source of protein alterations since both toxicants target the hepatic biliary canaliculus. Bile was collected before and after toxicant treatment. Biliary proteins were separated by one-dimensional SDS PAGE and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS-MS) with data-dependent scanning. Comprehensive analysis of biliary proteins was performed by using SEQUEST and BLAST database searching, in combination with de novo interpretation. Bile not subjected to tryptic digestion was analyzed for DCE metabolites. DCE treatment resulted in a marked increase in the overall number of biliary proteins, whereas few changes in the proteomic profile were apparent in bile after diclofenac treatment. This is consonant with prior observations of more profound effects of DCE on canalicular membrane integrity. LC-MS-MS analyses for DCE metabolites revealed the presence of S-carboxymethyl glutathione, S (cysteinylacetyl)glutathione, and a product of the intramolecular rearrangement of the DCE metabolite, ClCH(2)COSG, not previously described in vivo. In addition, several S-carboxymethylated proteins were identified in bile from DCE treated animals. This investigation has produced the first comprehensive baseline characterization of the content of the rat biliary proteome and the first documentation of alterations in the proteome of bile by toxicant treatment. In addition, the results provide direct in vivo evidence for DCE metabolic routes proposed in the formation of covalent adducts. PMID- 14565773 TI - 3D-MEDNEs: an alternative "in silico" technique for chemical research in toxicology. 1. prediction of chemically induced agranulocytosis. AB - A novel approach to molecular negentropy from the point of view of Markov models is introduced. Stochastic negentropies (MEDNEs) are used to develop a linear discriminant analysis. The discriminant analysis produced a set of two discriminant functions, which gave rise to a very good separation of 93.38% of 151 chemicals (training series) into two groups. The total predictability (86.67%, i.e., 52 compounds out of 60) was tested by means of an external validation set. Randic's orthogonalization procedures allowed interpretation of the model while avoiding collinearity descriptors. On the other hand, factor analysis was used to suggest the relation of MEDNEs with other molecular descriptors and properties into a property space. Three principal factors (related to three orthogonal MEDNEs) can be used to explain approximately 90% of the variance of different molecular parameters of halobenzenes including bulk, energetic, dipolar, molecular surface-related, and hydrophobic parameters. Finally, preliminary experimental results coincide with a theoretical prediction when agranulocytosis induction by G-1, a novel microcidal that presents Z/E isomerism, is not detected. PMID- 14565774 TI - DNA adduct formation from acrylamide via conversion to glycidamide in adult and neonatal mice. AB - Acrylamide (AA) is a high production volume chemical with many industrial uses; however, recent findings of ppm levels in starchy foods cooked at high temperature have refocused worldwide attention on the neurotoxicity, germ cell mutagenicity, and carcinogenicity of AA. Oxidative metabolism of AA to its epoxide metabolite, glycidamide (GA), has been observed in experimental animals and humans and may be associated with many of the toxic effects of AA exposure, including formation of N7-(2-carbamoyl-2-hydroxyethyl)guanine (N7-GA-Gua) in vivo. This paper describes the characterization of two new GA-derived DNA adducts formed in vitro, N3-(2-carbamoyl-2-hydroxyethyl)adenine (N3-GA-Ade) and N1-(2 carboxy-2-hydroxyethyl)-2'-deoxyadenosine. A sensitive method for quantification of N7-GA-Gua and N3-GA-Ade, based on LC with tandem mass spectrometry and isotope dilution, was developed and validated for use in measuring DNA adduct formation in selected tissues of adult and whole body DNA of 3 day old neonatal mice treated with AA and GA. In adult mice, DNA adduct formation was observed in liver, lung, and kidney with levels of N7-GA-Gua around 2000 adducts/10(8) nucleotides and N3-GA-Ade around 20 adducts/10(8) nucleotides. Adduct levels were modestly higher in adult mice dosed with GA as opposed to AA; however, treatment of neonatal mice with GA produced 5-7-fold higher whole body DNA adduct levels than with AA, presumably reflective of lower oxidative enzyme activity in newborn mice. DNA adduct formation from AA treatment in adult mice showed a supralinear dose-response relationship, consistent with saturation of oxidative metabolism at higher doses. These results increase our understanding of the mutagenic potential of GA and provide further evidence for a genotoxic mechanism in AA carcinogenesis. PMID- 14565776 TI - Bioactivation of clozapine by murine cardiac tissue in vivo and in vitro. AB - Clozapine, an atypical neuroleptic, undergoes bioactivation to a chemically reactive nitrenium ion. This has been implicated in the pathogenesis of clozapine induced agranulocytosis. Clozapine also causes myocarditis and cardiomyopathy, the mechanisms of which are unknown. To investigate this, we have evaluated whether clozapine undergoes bioactivation by murine cardiac tissue, in comparison to hepatic tissue. Mice were administered clozapine (5 and 50 mg/kg i.p.), and the extent of covalent binding was assessed by Western blotting. There was an increase in irreversible binding of clozapine to several proteins, ranging in mass from 30 to 250 kDa in both hepatic and cardiac tissue. Bioactivation by hepatic and cardiac microsomes was assessed by LC/MS using glutathione to trap the intermediate. Metabolism of radiolabeled clozapine to a glutathionyl conjugate by liver and cardiac microsomes was 30.5 +/- 3.3 and 3.6 +/- 0.3% of the initial incubation concentration, respectively. Ketoconazole (20 muM), a P450 inhibitor, significantly reduced binding in both hepatic and cardiac microsomes to 6.2 +/- 0.2 and 0.5 +/- 0.06%, respectively. These data indicate that clozapine undergoes bioactivation in the heart to a chemically reactive nitrenium metabolite that may be important in the pathogenesis of myocarditis and cardiomyopathy observed in man. PMID- 14565775 TI - Study of 202 natural, synthetic, and environmental chemicals for binding to the androgen receptor. AB - A number of environmental and industrial chemicals are reported to possess androgenic or antiandrogenic activities. These androgenic endocrine disrupting chemicals may disrupt the endocrine system of humans and wildlife by mimicking or antagonizing the functions of natural hormones. The present study developed a low cost recombinant androgen receptor (AR) competitive binding assay that uses no animals. We validated the assay by comparing the protocols and results from other similar assays, such as the binding assay using prostate cytosol. We tested 202 natural, synthetic, and environmental chemicals that encompass a broad range of structural classes, including steroids, diethylstilbestrol and related chemicals, antiestrogens, flutamide derivatives, bisphenol A derivatives, alkylphenols, parabens, alkyloxyphenols, phthalates, siloxanes, phytoestrogens, DDTs, PCBs, pesticides, organophosphate insecticides, and other chemicals. Some of these chemicals are environmentally persistent and/or commercially important, but their AR binding affinities have not been previously reported. To the best of our knowledge, these results represent the largest and most diverse data set publicly available for chemical binding to the AR. Through a careful structure-activity relationship (SAR) examination of the data set in conjunction with knowledge of the recently reported ligand-AR crystal structures, we are able to define the general structural requirements for chemical binding to AR. Hydrophobic interactions are important for AR binding. The interaction between ligand and AR at the 3- and 17-positions of testosterone and R1881 found in other chemical classes are discussed in depth. The SAR studies of ligand binding characteristics for AR are compared to our previously reported results for estrogen receptor binding. PMID- 14565777 TI - Development of QSARs for predicting the joint effects between cyanogenic toxicants and aldehydes. AB - Quantitative structure-activity relationship (QSAR) approaches are proposed in this study to predict the joint effects of mixture toxicity. The initial investigation studies the joint effects between cyanogenic toxicants and aldehydes to Photobacterium phosphoreum. Joint effects are found to result from the formation of a carbanion intermediate produced through the chemical interactions between cyanogenic toxicants and aldehydes. Further research indicates that the formation of carbanion intermediate is highly correlated with not only the charge of the carbon atom in the -CHO of aldehydes but also the charge of the carbon atom (C) in the carbochain of cyanogenic toxicants. The charge of the carbon atom in the -CHO of aldehydes is quantified by using the Hammett constant (sigma(p)), and then, sigma(p)-based QSAR models are proposed to describe the relationships between the joint effects and the chemical structures of the aldehydes. By using the charge of carbon atom (C) in the carbochain of cyanogenic toxicants, another QSAR model is proposed to describe the relationship between the joint effects and the chemical structures of cyanogenic toxicants. PMID- 14565778 TI - The parkinsonian neurotoxin 1-methyl-4-phenylpyridinium (MPP(+)) mediates release of l-3,4-dihydroxyphenylalanine (l-DOPA) and inhibition of l-DOPA decarboxylase in the rat striatum: a microdialysis study. AB - Reactive oxygen species (ROS) and reactive nitrogen species (RNS), particularly peroxynitrite, have been implicated as key participants in the dopaminergic neurotoxicity of 1-methyl-4-phenylpyridinium (MPP(+)). However, on the basis of available information, it is not clear whether the MPP(+)-induced overproduction of ROS and RNS occurs in the intraneuronal and/or extracellular compartment. Early steps in the neurotoxic mechanism evoked by MPP(+) include a profound dopaminergic energy impairment, which mediates a massive release of dopamine (DA), glutathione (GSH), and cysteine (CySH). In the event that MPP(+) mediates extracellular generation of ROS (such as superoxide and/or hydroxyl radicals) and/or peroxynitrite, released DA, GSH, and CySH should be oxidized forming thioethers of DA and disulfides. Using microdialysis experiments in which MPP(+) was perfused into the striatum of awake rats, the present study was unable to detect the presence of such biomarkers of extracellular ROS and/or RNS generation. However, MPP(+) induced a transient, concentration-dependent rise of extracellular l-3,4-dihydroxyphenylalanine (l-DOPA), identified on the basis of dialysate analysis using several HPLC methods and its conversion to DA by purified l-DOPA decarboxylase (DDC). Methamphetamine (30 mg/kg, i.p.) similarly caused a significant but transient rise of l-DOPA in the rat striatum. Antioxidants such as salicylate and mannitol had no effect on the MPP(+)-mediated elevation of extracellular l-DOPA, suggesting that it is not formed by nonenzymatic hydroxylation of l-tyrosine by ROS or RNS. Rather, in vivo, but not in vitro, MPP(+) caused rapid inhibition of DDC, which appears to result in intraneuronal accumulation and subsequent release of l-DOPA. Because l-DOPA can mediate l-glutamate release, as well as be an excitotoxin, the possibility is raised that l-DOPA may play a role in the dopaminergic neurotoxicity of MPP(+). PMID- 14565779 TI - Synthesis, properties, and NMR studies of a C8-phenylguanine modified oligonucleotide that preferentially adopts the Z DNA conformation. AB - Carcinogenic aryl hydrazines produce C8-arylated purine adducts. The effect of these adducts on DNA conformation and their role in hydrazine carcinogenesis are unknown. Here, we describe a new synthetic route to produce these adducts that is also compatible with the synthesis of the corresponding phosphoramidites needed for oligonucleotide synthesis. Two oligonucleotides were prepared, an unmodified oligonucleotide, d((5)(')CGCGCGCGCG(3)(')), and a C8-phenylguanine modified oligonucleotide, d((5)(')CGCGCGCGCG(3)(')) (G = 8-phenylguanine). These oligonucleotides were compared using thermal denaturation, circular dichroism, NMR, and molecular modeling. The phenyl modification destabilizes the B DNA form and stabilizes the Z DNA form such that the B:Z ratio is near one under physiological conditions. In light of recent studies that show a role for Z DNA in gene expression and cell transformation, Z DNA stabilization by C8-arylguanine formation from aryl hydrazines may be relevant to their role in carcinogenesis. PMID- 14565780 TI - Inhaled corticosteroids in the long-term management of patients with chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a major problem in the elderly population, with approximately 10% of the population affected. Since COPD is an inflammatory disorder of the pulmonary system, corticosteroids might be expected to improve clinical outcomes of the disease. Data from large, well designed randomised clinical trials in which approximately one third of patients were > or =65 years of age indicate that inhaled corticosteroids do not modify the natural history of COPD, as measured by the rate of decline in forced expiratory volume in 1 second (FEV1). However, these same studies also suggest that corticosteroids reduce the frequency of clinical exacerbations by nearly a third (compared with placebo). This beneficial effect is particularly pronounced among those with an FEV1 less than 50% of the predicted value. Withdrawal of inhaled corticosteroids, on the other hand, leads to increased symptoms and elevates the risk of exacerbations by 50% above baseline levels. Patients' health-related quality of life is also improved by the use of inhaled corticosteroids. It is clear that inhaled corticosteroids elevate the risk of thrush, dysphonia and skin bruising by 2-fold compared with placebo. In addition, the sum of evidence suggests a modest deleterious effect for inhaled corticosteroids on bone mineral density, especially for formulations that have an increased rate of systemic absorption. However, the clinical evidence of this observation is uncertain. The effect of inhaled corticosteroids on fracture risk is controversial with some observational studies suggesting a possible association. Whether inhaled corticosteroids increase the risk of ophthalmic complications (cataracts and glaucoma) is also uncertain. In conclusion, the current evidence indicates that inhaled corticosteroid therapy produces short- and long-term clinical benefits in COPD patients with moderate-to-severe disease and should be used as adjunctive therapy for elderly patients with COPD who experience frequent exacerbations or have moderately reduced lung function. PMID- 14565781 TI - Diagnosis and management of panic disorder in older patients. AB - Panic disorder occurs less frequently in the elderly than in younger adults and rarely starts for the first time in old age. Panic attacks that begin in late life should prompt the clinician to conduct a careful search for a depressive disorder, physical illness or drugs that could be contributing to their presence. When panic attacks do occur in the elderly, the symptoms are qualitatively similar to those experienced by younger people. The elderly, however, may have fewer and less severe symptoms and exhibit less avoidant behaviour. As panic disorder is typically a chronic or recurrent condition, its management requires a long-term approach. With the exception of one descriptive pilot study, there have been no randomised controlled trials of the treatment of panic disorder in later life. Therefore, recommendations regarding the management of this disorder in the elderly must be extrapolated from research pertaining to younger patients. Selective serotonin reuptake inhibitors (SSRIs), tricyclic antidepressants, monoamine oxidase inhibitors, benzodiazepines and cognitive behavioural therapy are efficacious treatments for panic disorder. There are no consistent differences in efficacy between classes of medications or between pharmacotherapy and cognitive behavioural therapy. Furthermore, there are no reliable predictors of response to one type of treatment compared with another. Treatment selection, therefore, depends on an individual assessment of the risks and benefits of each type of treatment (taking into account comorbid psychiatric and physical conditions), patient preference, cost and the availability of therapists skilled in cognitive behavioural techniques. As a general rule, antidepressant medication is preferable to a benzodiazepine as a first-line treatment for panic disorder in the elderly, especially given the high level of comorbidity between panic disorder and depressive disorders. Of the antidepressants, an SSRI is recommended as the initial choice of treatment in older patients. Anxious patients frequently misattribute somatic symptoms of anxiety to adverse effects of medication. Adherence with treatment, therefore, can be enhanced by starting antidepressant medication at a low dosage so as to avoid initial exacerbation of anxiety (but then gradually increasing the dosage to the therapeutic range), frequent follow up during the first few weeks of treatment, discussion about potential adverse effects and addressing any other concerns the patient may have about taking medication. Given the delayed onset of action of antidepressant medication, the short-term use of adjunctive lorazepam in the first few weeks of treatment may be helpful in selected patients. PMID- 14565782 TI - Effects of olanzapine on lipid abnormalities in elderly psychotic patients. AB - INTRODUCTION: Recently concern regarding the cause or worsening of diabetes mellitus by some of the second-generation antipsychotics and their adverse affects on lipid metabolism has caused growing concern amongst physicians and patients. This study aims to assess these effects in elderly patients with schizophrenia. METHODS: In a prospective 6-month follow-up study of elderly inpatients experiencing an acute psychotic exacerbation and exposed to olanzapine for the first time, patients underwent physical and psychiatric assessments including: routine laboratory tests (including serum cholesterol and triglycerides levels), and bodyweight and clinical rating scale measurement. All tests and evaluations were performed at baseline and at the end of study. RESULTS: Twenty-one elderly patients with schizophrenia (15 women and six men) mean age 71.7 +/- 8.2 years were included. All were diagnosed according to Diagnostic and Statistical Manual of Mental Disorders (4th edition) as patients with schizophrenia or schizoaffective disorder. Mean duration of olanzapine treatment was 289 days (SD +/- 139) and the mean olanzapine dosage at the end of the study was 12.9 mg/day. At the end of the study, no significant change from baseline serum lipid levels were found for triglycerides (paired differences = 12.8 [SD +/- 38.5], 95% CI -30.3 to +4.7, t = -1.5, df = 20, p = 0.143) or cholesterol (paired differences = -9.0 [SD +/- 43.5], 95% CI = -28.8 to +10.8, t = -0.95, df = 20, p = 0.355). CONCLUSION: The association between olanzapine exposure and lipid abnormalities may not hold true for older patients. Larger studies with elderly patients are needed to support the present report. PMID- 14565784 TI - Dutasteride. AB - Dutasteride, a potent inhibitor of type 1 and 2 5alpha-reductase, reduced dihydrotestosterone levels by >90% in 85% of patients following 1 years' administration of oral dutasteride 0.5 mg/day. A combined analysis of three placebo-controlled clinical studies conducted in patients with benign prostatic hyperplasia (BPH) found sustained improvements in American Urological Association Symptom Index scores and urinary flow rate and a 57% decrease in the risk of acute urinary retention throughout the 2-year treatment period (all p < 0.001 vs placebo). Total prostate and transition zone volume were also reduced (both p < 0.001), as was the risk of BPH-related surgery (by 48%). A nonblind extension study found that dutasteride maintains efficacy for up to 4 years. Dutasteride monotherapy maintained symptom relief following combination treatment with dutasteride and tamsulosin in all patients but those with severe symptoms. Dutasteride was generally well tolerated. Impotence, reduced libido, gynaecomastia and ejaculation disorder occurred significantly more often in dutasteride than placebo recipients, but incidence was generally low. With the exception of gynaecomastia, incidence consistently decreased over time. PMID- 14565783 TI - Rationale for a trial of low-dose aspirin for the primary prevention of major adverse cardiovascular events and vascular dementia in the elderly: Aspirin in Reducing Events in the Elderly (ASPREE). AB - Low-dose aspirin (acetylsalicylic acid) therapy has been shown to reduce the risk of vascular events and there is increasing evidence of its potential to reduce the rate of cognitive decline in the elderly. Adverse effects including gastrointestinal and intracranial haemorrhage may offset these benefits. The balance of risks versus benefits of aspirin for the primary prevention of cardiovascular disease and vascular dementia has not been established in the elderly. There is clearly a need to conduct a study in family practice to investigate whether routine use of low-dose aspirin for the primary prevention of cardiovascular disease and vascular dementia in the elderly is beneficial or harmful. Aspirin in reducing events in the elderly (ASPREE) is a placebo controlled trial of low-dose aspirin for the primary prevention of major adverse cardiovascular events and vascular dementia. It will follow 15,000 subjects aged 70 years or more for an average of 5 years. This sample size has a power of 87% to detect a 15% reduction in primary events in the aspirin group, with an anticipated combined primary event rate of 20 per 1000 patient years. PMID- 14565788 TI - Gefitinib therapy for advanced non-small-cell lung cancer. AB - OBJECTIVE: To review the pharmacology, pharmacokinetics, clinical efficacy, and toxicity of gefitinib in non-small-cell lung cancer (NSCLC). DATA SOURCES: Primary literature search through MEDLINE and CANCERLIT, and abstract presentations (1966-May 2003). STUDY SELECTION AND DATA EXTRACTION: All published trials and abstracts citing gefitinib were evaluated, and all information deemed relevant was included in this article. DATA SYNTHESIS: NSCLC is known to overexpress epidermal growth factor receptor (EGFR). Gefitinib is a selective EGFR tyrosine kinase inhibitor. Based on the Phase I/II trial results, the optimal dose is 250 mg/d orally. It is well tolerated, with minimal and reversible toxicity. Skin rash and diarrhea are the most common adverse effects. Recent trials have shown that gefitinib provided a 10% tumor response rate and improved disease-related symptoms in patients with refractory, advanced NSCLC. CONCLUSIONS: Gefitinib, with a unique mechanism of action and favorable toxicity profile, has demonstrated clinical activity in NSCLC patients with chemotherapy refractory disease. It provides a valuable addition to the treatment options as monotherapy in patients with advanced NSCLC after failure of both platinum-based and docetaxel chemotherapies. Further research is required to evaluate the use of gefitinib in different clinical settings. PMID- 14565787 TI - Brinzolamide : a review of its use in the management of primary open-angle glaucoma and ocular hypertension. AB - Brinzolamide is a highly specific carbonic anhydrase (CA) inhibitor which lowers intraocular pressure (IOP) by reducing the rate of aqueous humour formation. Formulated as a 1% ophthalmic suspension (Azopt) and administered twice or three times daily, brinzolamide is indicated for the topical management of primary open angle glaucoma (POAG) and ocular hypertension (OH) as either monotherapy or adjunctive therapy with topical beta-blockers. As monotherapy in patients with POAG or OH, brinzolamide 1% demonstrated IOP-lowering efficacy that was significantly greater than placebo, equivalent to three-times-daily dorzolamide 2% but significantly lower than twice-daily timolol 0.5%. Brinzolamide 1% was equally effective in twice- and three-times-daily regimens producing diurnal mean IOP reductions from baseline in the range of 13.2-21.8%. When used adjunctively twice daily with timolol 0.5%, brinzolamide 1% was as effective as dorzolamide 2% and superior to placebo in lowering IOP in patients with POAG or OH. In clinical trials, brinzolamide 1% was well tolerated causing only nonserious adverse effects that were generally local, transient and mild to moderate in severity. The incidence of the most common adverse events associated with the use of brinzolamide 1% was either similar to (blurred vision and abnormal taste) or significantly lower than (ocular discomfort) with dorzolamide 2%. Topical brinzolamide 1% does not appear to produce the acid-base or electrolyte disturbances and severe systemic adverse effects characteristic of oral CA inhibitors. It can be used in patients unresponsive to beta-blockers or in whom beta-blockers are contraindicated. Brinzolamide 1% administered twice daily is among the least costly alternatives and adjuncts to beta-blocker therapy for glaucoma and is generally associated with less direct medical cost than dorzolamide. CONCLUSION: Brinzolamide 1% ophthalmic suspension administered twice or three times daily, as monotherapy or adjunctive therapy with topical beta blockers, has good IOP-lowering efficacy in patients with POAG or OH that is equivalent to that of dorzolamide 2% (three times daily as monotherapy, twice daily as adjunctive therapy). Brinzolamide is generally well tolerated and does not produce the systemic adverse effects associated with oral CA inhibitors. It can be used in patients who are unresponsive to, intolerant of, or unable to receive, ophthalmic beta-blockers. Thus, brinzolamide, either as monotherapy or adjunctive therapy with topical beta-blockers, should be regarded as a good second-line option in the pharmacological management of POAG and OH, and may be preferred over dorzolamide because of significantly less ocular discomfort. PMID- 14565792 TI - Venlafaxine treatment of fibromyalgia. AB - BACKGROUND: Although the pathophysiology of fibromyalgia is unknown, central monoaminergic transmission may play a role. Antidepressants have proved to be successful in alleviating symptoms of fibromyalgia. Medications that act on multiple neurotransmitters may be more effective in symptom management. OBJECTIVE: To assess the efficacy of venlafaxine, a potent inhibitor of both norepinephrine and serotonin reuptake, in the treatment of patients with fibromyalgia. METHODS: Fifteen patients with fibromyalgia were assessed prior to and after treatment with fixed-dose venlafaxine 75 mg/d. Before initiation of pharmacotherapy, patients were interviewed with the Structured Clinical Interview for Axis I disorders in the Diagnostic and Statistical Manual of Mental Disorders, 4th edition. The study lasted for 12 weeks, and patients were evaluated in weeks 6 and 12. The primary outcome measures were the Fibromyalgia Impact Questionnaire (FIQ) total score and pain score. The anxiety and depression levels of the patients were measured with the Beck Depression, the Beck Anxiety, the Hamilton Anxiety, and the Hamilton Depression scales. RESULTS: There was a significant improvement in the mean intensity of pain (F = 14.3; p = 0.0001) and in the disability caused by fibromyalgia (F = 42.7; p = 0.0001) from baseline to week 12 of treatment. The depression and anxiety scores also decreased significantly from baseline to week 12. The improvement in the FIQ scores did not correlate with the decrease of scores in both patient- and physician-rated depression and anxiety inventories. Change in pain scores also was not correlated with the change in depression and anxiety scores. CONCLUSIONS: Venlafaxine was quite promising in alleviating the pain and disability associated with fibromyalgia. This effect seems to be independent of its anxiolytic and antidepressant properties. Blockade of both norepinephrine and serotonin reuptake might be more effective than blockade of either neurotransmitter alone in the treatment of fibromyalgia. PMID- 14565793 TI - Treatment of impending tumor lysis with single-dose rasburicase. AB - OBJECTIVE: To report the experience of using rasburicase as a single-dose treatment for childhood leukemia presenting with hyperuricemia. CASE SUMMARIES: Three children with acute lymphoblastic leukemia presenting with hyperuricemia received rasburicase as a single intravenous dose just prior to the start of chemotherapy. This was followed by rapid reduction of serum uric acid levels within 24 hours, which remained low throughout induction therapy while allopurinol and hydration therapy without urinary alkalinization ensued. Subclinical tumor lysis was evidenced by the appearance of hyperphosphatemia and hypocalcemia in all cases and hyperkalemia in 1 patient. These abnormalities were transient, and each patient's renal function gradually improved from pretreatment baseline without requiring dialysis. DISCUSSION: Our experience suggests that hyperuricemia in children at risk for tumor lysis can be managed with a briefer regimen of rasburicase than the recommended 5- to 7-day course. CONCLUSIONS: A shorter course of rasburicase treatment, including single-dose injection, is feasible and will improve the cost-effectiveness profile of the otherwise expensive compound. PMID- 14565794 TI - Causes and management of hyponatremia. AB - OBJECTIVE: To review clinical information on the incidence and causes of hyponatremia (defined as a serum sodium level <130 mEq/L), the most common electrolyte abnormality seen in general hospital patients, and to discuss the diagnosis and treatment of hyponatremia in relation to these factors. DATA SOURCES: Primary sources and review articles were identified via MEDLINE (1981 July 2003) for entries on hyponatremia. We limited the search to specific topics including incidence, risk factors, diagnosis, treatment, and clinical disorders and medications associated with hypotonic hyponatremia. STUDY SELECTION AND DATA EXTRACTION: All of the articles identified were evaluated, and relevant and representative information was included in this review. DATA SYNTHESIS: Hyponatremia can result from several disease states, injury, surgery, physical exercise, or the administration of certain drugs (e.g., antidepressants, antiepileptics) and is associated with advanced age. Drug-induced hyponatremia is often asymptomatic and usually resolves following water restriction and monitoring of medication. Symptoms of hyponatremia are primarily neurologic; the principal danger of hyponatremia relates to effects on central nervous system function due to changes in brain size. CONCLUSIONS: Although hyponatremia can be a serious condition, appropriate measures for the management of at-risk and affected patients will lead to full recovery in most cases. PMID- 14565795 TI - Multivitamin supplements may affect warfarin anticoagulation in susceptible patients. AB - OBJECTIVE: To report an interaction of a multivitamin preparation containing small amounts of vitamin K(1) (25 microg) with warfarin in a case series and to assess the prevalence of vitamin K(1) deficiency in ambulatory anticoagulated patients. CASE SUMMARIES: We describe 3 patients whose anticoagulation was stabilized with warfarin in whom initiation or cessation of a self-prescribed multivitamin supplement delivering 25 microg of vitamin K(1) daily was associated with an otherwise unexplained significant fall or rise in international normalized ratio (INR), respectively, with major thrombosis or hemorrhage in 2. This interaction was rated probable on the Naranjo probability scale. Suspecting vitamin K(1) deficiency as an explanation for this oversensitivity, we assessed the prevalence of vitamin K(1) deficiency in our clinic by determining plasma vitamin K(1) levels in 179 stable consecutive patients, finding very low levels (<0.1 ng/mL) in 22 of 179 (12%). DISCUSSION: Vitamin K(1) supplements of 25 microg daily are far below the dose thought to affect anticoagulant control. We hypothesize that, in our patients, unsuspected vitamin K(1) deficiency caused an oversensitivity to small vitamin K(1) supplements. In patients with low vitamin K(1) status, even such low doses represent a significant increment in daily intake, thus lowering the sensitivity to warfarin. Our analysis suggests that low vitamin K(1) status exists in a small, but important, minority of ambulatory patients undergoing anticoagulation. CONCLUSIONS: Clinicians should instruct anticoagulated patients to report the use of multivitamin supplements and inquire about it in cases of unexplained INR changes. PMID- 14565796 TI - Prolonged thrombocytopenia associated with isotretinoin. AB - OBJECTIVE: To report a case of severe prolonged thrombocytopenia possibly associated with isotretinoin. CASE SUMMARY: A 27-year-old white woman developed severe thrombocytopenia and elevated transaminases after 3(1/2) months of treatment with isotretinoin. Prior to the onset of thrombocytopenia, the patient had also received a 10-day course of cephalexin. Rectal bleeding was reported by the patient, who was otherwise asymptomatic. Liver enzyme values returned to normal approximately 1 week after discontinuation of isotretinoin; however, platelet counts required approximately 2 months to normalize. Based on the Naranjo probability scale, possible causality exists between isotretinoin and thrombocytopenia. DISCUSSION: The exact mechanism by which isotretinoin caused thrombocytopenia in this patient is not clearly understood. To our knowledge, only 3 previous cases of isotretinoin-associated thrombocytopenia have been reported. The long recovery process that occurred in our patient is possibly a direct result of the long elimination half-life of both the parent compound and active metabolites of isotretinoin. CONCLUSIONS: Clinicians prescribing isotretinoin should be aware of the potential life-threatening consequence of thrombocytopenia, and a complete blood cell count with platelets should be part of the routine monthly monitoring in all patients receiving isotretinoin therapy. PMID- 14565798 TI - Functional health literacy and medication use: the pharmacist's role. PMID- 14565797 TI - Estradiol in severe asthma with premenstrual worsening. AB - OBJECTIVE: To describe the beneficial effects of estradiol in a severely asthmatic woman with premenstrual worsening of asthma. CASE SUMMARY: A 50-year old white woman, with a 14-year history of severe steroid-dependent asthma and monthly premenstrual worsening of asthma, was randomized to receive estradiol 2 mg or placebo for 6 days during the late luteal phase (days 23-28) of 2 successive menstrual cycles. Despite greater prednisone and inhaled beta-agonist use during the late luteal phase of the placebo cycle, the patient exhibited improved asthma symptoms, pulmonary function, and peak expiratory flows, as well as lower values for biomarkers of airway inflammation during the same time period of the estradiol cycle. DISCUSSION: This subject's response to estradiol was in contrast to results of a randomized, controlled trial involving patients with predominantly mild asthma under excellent control. The discrepant observations for this case versus the majority of subjects in the randomized, controlled study suggest that premenstrual asthma (PMA) and a beneficial response to estradiol may be more likely in women with severe asthma. CONCLUSIONS: The administration of estradiol during the late luteal phase of the menstrual cycle to a woman with severe asthma with PMA was associated with improved asthma symptoms, pulmonary function, and peak expiratory flows, and lower serum eosinophil protein X and urinary leukotriene E(4) biomarker concentrations. Thus, estradiol's potential role in women with severe asthma and PMA may warrant further exploration in this subgroup. PMID- 14565799 TI - Enteric-coated mycophenolate sodium. AB - OBJECTIVE: To review the pharmacology, pharmacokinetics, efficacy, and safety of mycophenolate sodium. DATA SOURCES: Primary literature was obtained via a MEDLINE search (1966-June 2003). Abstracts were obtained from the manufacturer and included in the analysis. STUDY SELECTION AND DATA EXTRACTION: All studies and abstracts evaluating mycophenolate sodium in solid organ transplantation were considered for inclusion. English-language studies and abstracts were selected for inclusion, but were limited to those consisting of human subjects. DATA SYNTHESIS: Mycophenolate sodium, a mycophenolic acid prodrug, is an inhibitor of T-lymphocyte proliferation. Mycophenolic acid reduces the incidence of acute rejection in renal transplantation. Mycophenolate sodium is enteric coated and has been suggested as a potential method to reduce the gastrointestinal adverse events seen with mycophenolate mofetil. Both mycophenolate mofetil and mycophenolate sodium have been shown to be therapeutically equivalent at decreasing the incidence of allograft rejection and loss. The frequency of adverse events is similar between both compounds, with the most common events being diarrhea and leukopenia. CONCLUSIONS: Mycophenolate sodium is effective in preventing acute rejection in renal transplant recipients. At doses of 720 mg twice daily, the efficacy and safety profiles are similar to those of mycophenolate mofetil 1000 mg twice daily. Mycophenolate sodium has been approved in Switzerland; approval in the US is pending. PMID- 14565800 TI - Drug-induced esophageal injuries and dysphagia. AB - OBJECTIVE: To review and analyze medical literature documenting drug-induced esophageal injury and dysphagia and to formulate strategies to enhance pharmacists' prevention, detection, and treatment of these iatrogenic complications. DATA SOURCES: A MEDLINE search (1966-April 2002) was conducted to identify primary and secondary literature using variable combinations of the following search terms: pill-induced, drug-induced, or iatrogenic with esophageal injury, esophageal damage, or dysphagia. Bibliographies were also reviewed to identify additional relevant references. STUDY SELECTION AND DATA EXTRACTION: All case reports, reviews, and clinical studies relating to drug-induced esophageal injury or swallowing dysfunction were evaluated. DATA SYNTHESIS: Drug-induced esophageal injury may be under-recognized. Several drugs have been associated with physical or chemically mediated injuries. Risk factors for injury have been identified and preventive and treatment strategies have been successful in limiting esophageal injury. Drug-induced dysphagia can have serious complications and is most often associated with typical neuroleptics such as haloperidol. CONCLUSIONS: Pharmacists can play a pivotal role in proactively identifying situations where there is a higher likelihood of drug-induced esophageal injury or dysphagia. They can recommend preventive strategies to promote safe medication use, help identify iatrogenic complications when they occur, and assist in formulation of appropriate treatment strategies. PMID- 14565802 TI - Evaluation of HIV drug interaction web sites. AB - BACKGROUND: Clinicians frequently consult HIV drug interaction Web sites of unknown quality. OBJECTIVE: To systematically review and identify HIV drug interaction Web sites of high quality and usefulness for healthcare professionals. METHODS: Relevant Web sites were identified through a structured search on commonly used search engines. An assessment tool containing 4 domains (content, reliability, access restrictions, ease of navigation) was developed. English and French Web sites were selected for review if they included HIV drug interaction information directed to healthcare professionals. Web sites were excluded if antiretroviral interaction data were not available or were out of date. Commercial online databases and sites that required payment were not included. Seventeen HIV pharmacists from across Canada participated in the review. The Web sites were ranked with total mean scores. Mean scores for each domain were then analyzed. Interrater agreement and ANOVA using the rater as a covariate were determined. RESULTS: Nine Web sites met the criteria for review. Web sites from Toronto General Hospital (Canada), HIVinSite (beta version) (US), and the University of Liverpool (UK) ranked highest for total mean scores and for content. Other Web sites were found to be reliable, accessible, and easy to navigate; however, they did not consistently include unpublished data or data on herbal preparations, recreational drugs, or multiple interactions. CONCLUSIONS: Three HIV interaction Web sites of high quality were identified that can be valuable tools for HIV and non-HIV health-care professionals. Regular reviews are necessary in order to keep pace with the growing body of HIV interaction data and the constant evolution of Web sites. PMID- 14565801 TI - Treatment of pediatric idiopathic pulmonary hemosiderosis with low-dose cyclophosphamide. AB - OBJECTIVE: To report the safety and efficacy of long-term, low-dose cyclophosphamide therapy in a child with idiopathic pulmonary hemosiderosis (IPH). CASE SUMMARY: A 7-year-old boy diagnosed with IPH 4 years previously was initially prescribed prednisolone. Because he only had a transient response to prednisolone, oral cyclophosphamide 2 mg/kg/d was later added. A dramatic improvement was noted during the subsequent follow-up. One year after cyclophosphamide therapy, the patient suddenly developed thrombocytopenia (platelet count 75 x 10(3)/mm(3)), with the platelet count decreasing to 10 x 10(3)/mm(3) over the following 10 months. Cyclophosphamide was tapered to an alternating daily dosage of 1 mg/kg. The tapering resulted in a subsequent increase in the platelet count, which was maintained between 20 and 50 x 10(3)/mm(3) without occurrence of petechiae or spontaneous bleeding. Under this reduced dosing regimen, the disease has remained in remission for >1 year. DISCUSSION: Due to the low prevalence of IPH, only limited data document the safety and efficacy of immunosuppressive therapy in treating this disease. Although our patient showed a good response to low-dose cyclophosphamide, he developed thrombocytopenia with its use. The mechanism is unclear, but it may be similar to that of high-dose cyclophosphamide-induced myelosuppression. Due to the development of thrombocytopenia, the use of cyclophosphamide was maintained under a reduced dosing regimen. The benefit of long-term immunosuppressive therapy is controversial, and more clinical evidence is required to support its continued usage. CONCLUSIONS: Long-term, low-dose cyclophosphamide is effective in treating childhood IPH, but caution should be exercised due to the possible development of thrombocytopenia. Periodic monitoring of the platelet count in long-term treatment is recommended. PMID- 14565803 TI - Leukocytoclastic vasculitis related to rofecoxib. PMID- 14565804 TI - C-reactive protein, antiinflammatory drugs, and quality of life in diabetes. AB - BACKGROUND: C-reactive protein (CRP) is an important biological marker of inflammation that has been linked to cardiovascular disease. The extent to which the inflammatory processes associated with elevated CRP concentrations impair physical functioning and quality of life, and whether this is modulated by hydroxymethylglutaryl coenzyme A reductase inhibitors (statins) and aspirin, which have been shown to lower CRP concentrations, is unclear. OBJECTIVE: To determine whether an association exists between CRP concentrations and health related quality of life among patients with diabetes, and to determine whether the association is affected by use of statins and nonsteroidal antiinflammatory drugs (NSAIDs). METHODS: A trained interviewer collected self-reported information regarding demographics, health-related quality of life (SF-12), and medication use, and a nurse collected a blood sample from 86 adult subjects with type 2 diabetes mellitus who were part of a larger population-based survey. The serum was analyzed for CRP using a highly sensitive assay. RESULTS: In simple bivariate analysis, there was a significant inverse relationship between CRP and the physical health component score of health-related quality of life (Spearman correlation coefficient [r] = -0.26; p = 0.025). For subjects on statins (r = 0.44; p = 0.02; n = 27), this relationship persisted, while for patients on NSAIDs or aspirin (r = -0.21; p = 0.17; n = 44), no relationship was observed. Similar findings were observed with self-rated health alone as an outcome variable. However, stepwise linear regression revealed no consistent relationship between CRP and health-related quality of life (i.e., standardized SF-12 physical component score) when demographic variables, disease duration, glucose control, serum creatinine, smoking, and medication use were controlled for. CONCLUSIONS: In multivariate regression modeling, the initial inverse relationship between CRP and health-related quality of life was lost, suggesting that other covariates are responsible for this association. While statins and NSAIDs may impact CRP or health-related quality of life independently, they do not appear to modulate a relationship between these factors. PMID- 14565805 TI - Primary prevention of coronary heart disease in the elderly. AB - OBJECTIVE: To review relevant literature supporting the use of antihypertensive agents, lipid-lowering agents (i.e., statins), and aspirin therapy for the primary prevention of coronary heart disease (CHD) in an elderly patient population (age >or=65 y). DATA SOURCES: A MEDLINE search (1988-January 2003) was conducted. STUDY SELECTION AND DATA EXTRACTION: Primary and tertiary literature involving the uses of antihypertensives, statins, and aspirin therapy in the elderly were reviewed. DATA SYNTHESIS: Mortality due to CHD in the US population has decreased 40-50% over the last 30 years; however, CHD remains the leading cause of morbidity and mortality in elderly persons. As the population continues to age, the number of older adults eligible for primary prevention will rise. The American Heart Association clinical practice guidelines for the primary prevention of CHD were updated in 2002; however, they are based on findings from clinical trials that enrolled predominantly middle-aged white men. The recommendations for elderly individuals are predominantly extrapolated from subgroup analyses of randomized clinical trials or cohort studies. This literature suggests that elderly persons are candidates for primary prevention measures and experience reductions in coronary events when treated with appropriate therapies. CONCLUSIONS: Data suggest that use of antihypertensives, statins, and aspirin therapy in the elderly appears effective to an extent similar to, and often greater than, that observed in younger patients. We believe these agents should be prescribed to all appropriate high-risk elderly patients. Ongoing and future studies will more clearly elucidate the benefits of primary prevention therapy, particularly in persons >or=75 years of age. PMID- 14565806 TI - Allopurinol as a cardioprotectant during coronary artery bypass graft surgery. AB - OBJECTIVE: To examine the role of allopurinol as a cardioprotectant during coronary artery bypass graft (CABG) surgery. DATA SOURCES: A search of MEDLINE (1966-October 2002) was performed using the following terms: allopurinol, xanthine oxidase, oxygen free radical, and coronary artery bypass. References evaluated were limited to English-language and human studies, yielding 41 citations, 13 of which were found suitable. The 5 largest studies are discussed. DATA SYNTHESIS: Multiple studies with various doses have evaluated the effects of allopurinol on outcomes in CABG patients. These studies found that allopurinol can reduce in hospital mortality, improve cardiac performance, reduce incidence of arrhythmias, reduce markers of ischemia and free-radical generation, and reduce the need for inotropic support. However, these findings were not consistent between all studies. CONCLUSIONS: Allopurinol may reduce the incidence of CABG complications. Although the optimal dose has not been determined, reviewed literature suggests that patients should receive at least 600 mg one day prior to surgery, as well as at least 600 mg on the day of surgery. PMID- 14565807 TI - Oral terbinafine-induced bullous pemphigoid. AB - OBJECTIVE: To report a case of drug-induced bullous pemphigoid (BP) in an otherwise healthy elderly patient after taking terbinafine for 20 days. CASE SUMMARY: A 78-year-old man presented with distal subungual onychomycosis of the toenails. Oral terbinafine 250 mg/d was started for treatment. Twenty days after the beginning of the treatment, the patient presented with widespread pruritic blistering eruption, more severe on the extremities. He had not received terbinafine before and was not receiving any other drug. The diagnosis of BP was confirmed with histopathologic examination. The patient was treated successfully with oral steroids. DISCUSSION: Drug-induced BP is a well-known complication of many drugs and represents a spectrum from an acute and self-limited condition to a chronic disease. Terbinafine is frequently used in the elderly. It is known to have less drug interactions and adverse effects compared with the other antifungal agents. Use of the Naranjo probability scale indicated a probable relationship between BP and terbinafine treatment in this patient. CONCLUSIONS: To the best of our knowledge, this is the first report in the English-language literature of BP in a patient taking terbinafine. As with any other medication, oral terbinafine has the potential for severe adverse reactions. Healthcare professionals should be aware of this possibility. PMID- 14565808 TI - Safety and efficacy of influenza vaccine in children. AB - OBJECTIVE: To describe the safety and efficacy of influenza vaccines in asthmatic children. DATA SOURCES: Literature was identified by a MEDLINE search (2002-March 2003). Key search terms included asthma, exacerbation, children, vaccine, and influenza. DATA SYNTHESIS: Concerns that the influenza vaccine may exacerbate asthma attacks have kept many asthmatic children from receiving this immunization. Researchers have conducted studies to determine the burden of influenza on asthmatic children, the safety of influenza vaccines, and their benefit in the presence of glucocorticoid burst therapy in the same population. CONCLUSIONS: Influenza vaccines tested are safe and efficacious in asthmatic children. PMID- 14565809 TI - Cardiorespiratory effects of naloxone in children. AB - BACKGROUND: Data on the cardiorespiratory changes and complications following administration of naloxone in children are limited. OBJECTIVE: To evaluate the cardiorespiratory changes and complications following naloxone treatment in children. METHODS: The maximal changes in respiratory rate (RR), heart rate (HR), systolic (SBP) and diastolic (DBP) blood pressure, and any complications within 1 and 2 hours following naloxone were tabulated. RESULTS: One hundred ninety-five children received naloxone over 3 years. The mean +/- SD age was 9.7 +/- 6 years. The total doses of naloxone ranged from 0.01 to 7 mg (0.001-0.5 mg/kg body weight), with a median dose of 0.1 mg. Group 1 patients consisted of 116 (60%) children who were postoperative and had been given naloxone by an anesthesiologist; group 2 patients consisted of 79 (40%) children who received naloxone in the emergency department or pediatric intensive care unit. Patients in group 1 were older: 10.6 +/- 5.3 versus 8.2 +/- 6.7 years (p < 0.006), but received significantly lower doses of naloxone (0.09 +/- 0.2 vs. 1.1 +/- 0.76 mg; p < 0.001). When the entire cohort was evaluated, a significant increase in RR (15 +/- 7 vs. 21 +/- 8 breaths/min; p < 0.001), HR (102 +/- 29 vs.107 +/- 29 beats/min; p < 0.001), SBP (109 +/- 17 vs. 115 +/- 15 mm Hg; p < 0.001), and DBP (56 +/- 10 vs. 60 +/- 13 mm Hg; p < 0.001) within 1 hour following naloxone was noted. When the 2 groups were compared, only the changes in RR were greater in group 2 patients (6.8 +/- 7.9 vs. 4.7 +/- 5 breaths/min; p < 0.001) following naloxone. Systolic hypertension occurred in 33 of 195 (16.9%) of all patients, while diastolic hypertension occurred in 13 (6.6%) of all patients after naloxone. Only the incidence of diastolic hypertension was higher in group 2 compared with group 1 patients following naloxone (16% vs. 2%; p < 0.001). Hypertension resolved spontaneously. One child developed pulmonary edema and required positive pressure ventilation for 22 hours. CONCLUSIONS: Moderate increases in RR, HR, and BP occur after naloxone administration to children, but development of more serious complications is rare. PMID- 14565810 TI - Efficacy of sulfonylureas with insulin in type 2 diabetes mellitus. AB - BACKGROUND: In subjects with type 2 diabetes mellitus, glycemic control deteroriates while patients use sulfonylurea drugs during the course of the disease. Adjunctive therapy with insulin at this stage requires a lesser daily insulin dose in comparison with insulin monotherapy while restoring desirable glycemic control. However, data regarding direct comparison between various sulfonylureas in this regard are lacking. OBJECTIVE: To examine comparative efficacies of adjunctive therapy with insulin in subjects with type 2 diabetes manifesting lapse of glycemic control while receiving various individual sulfonylurea drugs. METHODS: Four groups of 10 subjects, each presenting with glycosylated hemoglobin (HbA(1C)) >8.0% while using either tolazamide, glyburide, glipizide Gastrointestinal Therapeutic System (GITS), or glimepiride, were recruited. Two from each group were randomized to receive placebo; the others continued the same drug. Pre-supper subcutaneous 70 NPH/30 regular insulin was initiated at 10 units and gradually increased and adjusted as necessary to attain fasting blood glucose levels between 80 and 120 mg/dL and maintain the same range for 6 months. Fasting plasma glucose, plasma C-peptide, and HbA(1C) concentrations were determined prior to the addition of insulin and at the end of the study. Daily insulin dose and changes in body weight (BW) were noted at the end of the study, and the number of hypoglycemic events during the last 4 weeks of the study was determined. RESULTS: Daily insulin dose (units/kg BW), weight gain, and number of hypoglycemic events were significantly lower (p < 0.01) in subjects receiving sulfonylureas in comparison with placebo. However, the daily insulin dose alone was significantly lower (p < 0.05) with glimepiride (0.49 +/- 0.10; mean +/- SE) than with other sulfonylureas (tolazamide 0.58 +/- 0.12, glyburide 0.59 +/- 0.12, glipizide GITS 0.59 +/- 0.14). Finally, a significant correlation (r = 0.68; p < 0.001) was noted between suppression of plasma C peptide level and the daily insulin dose among all participants. CONCLUSIONS: By lowering the daily insulin dose, sulfonylurea drugs appear to improve the sensitivity of exogenous insulin in subjects with type 2 diabetes mellitus manifesting lapse of glycemic control. Moreover, glimepiride appears to possess a greater insulin-sparing property than other sulfonylureas. PMID- 14565811 TI - Use of NSAIDs for the chemoprevention of colorectal cancer. AB - OBJECTIVE: To discuss the role of nonsteroidal antiinflammatory drugs (NSAIDs) in the chemoprevention of colorectal cancer. DATA SOURCES: A MEDLINE search (1966 May 2003) was performed to identify key literature. Search items included, but were not limited to, NSAIDs, colorectal cancer, chemoprevention, cyclooxygenase-2 (COX-2)-specific inhibitors, and familial adenomatous polyposis (FAP). STUDY SELECTION AND DATA EXTRACTION: The search included experimental (in vitro and animal models) and clinical studies evaluating the use of NSAIDs for the chemoprevention of colorectal cancer. The MEDLINE search was supplemented by references from selected articles. DATA SYNTHESIS: Numerous experimental, epidemiologic, and clinical studies suggest that NSAIDs have promise as anticancer agents. The mechanism by which NSAIDs lead to decreased colon carcinogenesis is not fully understood, but may involve restoration of apoptosis and inhibition of prostaglandin-mediated angiogenesis. Compelling evidence from many observational studies has consistently documented a 40-50% reduction in the risk of adenomatous polyps, colorectal cancer incidence, and mortality in patients using NSAIDs. Recent randomized, controlled trials have demonstrated a benefit with aspirin in reducing the rate of development of new or recurrent adenomas in high-risk patients. In addition, randomized studies using sulindac and celecoxib in patients with FAP have documented significant regression of existing adenomatous polyps. CONCLUSIONS: Inhibition of COX-2 is an example of a targeted approach to the chemoprevention of colorectal cancer. However, controversy exists about the safety, efficacy, and optimal treatment regimen of NSAIDs as long-term chemopreventive agents in the general population. Ongoing studies in high-risk patients with both selective and nonselective COX inhibitors will provide important information in the area of colorectal chemoprevention, but clinical trials' use of adenomas as surrogate markers for chemoprevention trials makes their application to the general population limited. PMID- 14565812 TI - Venlafaxine hydrochloride for the treatment of hot flashes. AB - OBJECTIVE: To review the literature evaluating venlafaxine for the treatment of hot flashes. DATA SOURCES: Clinical literature accessed through MEDLINE (1966 August 2002), PubMed, Harrison's Online, and references of reviewed articles. Key terms used were venlafaxine, Effexor, hot flashes, and vasomotor symptoms. DATA SYNTHESIS: Not all patients experiencing hot flashes are candidates for traditional hormonal therapy. Nonhormonal alternatives have long been explored, but conflicting evidence of efficacy exists. CONCLUSIONS: Venlafaxine is an effective nonhormonal alternative for relief from uncontrolled hot flashes. PMID- 14565813 TI - Gender differences in NSAID use among older adults with osteoarthritis. AB - BACKGROUND: Nonsteroidal antiinflammatory drugs (NSAIDs) are the most commonly prescribed medications for the treatment of osteoarthritis (OA). Little is known about whether there are important gender differences in NSAID use among patients with OA. OBJECTIVE: To examine gender differences in patterns of NSAID use among older adults (>or=65 y) with OA. METHODS: Subjects (n = 11298) were members of a statewide prescription drug plan who responded to a health-related quality-of life (HRQOL) survey in 1997 and had a physician diagnosis of OA. Gender differences in patterns of NSAID use were examined over a 2-year period. RESULTS: Approximately one-third of the participants filled at least 1 NSAID prescription during the study. Women were significantly more likely to be prescribed an NSAID than men (37% vs. 30%), had a greater total days' supply of NSAIDs, and were more frequently prescribed NSAIDs with greater degrees of cyclooxygenase-2 selectivity. These gender differences persisted in statistical analyses controlling for demographic factors, HRQOL, and gastrointestinal (GI) risk factors. CONCLUSIONS: Results of this study showed significant gender differences in patterns of NSAID use, and these differences were independent of the risk for GI adverse effects and self-reported symptoms. Further research is needed to examine reasons for these gender variations, as well as their impact on the quality of symptom management. PMID- 14565814 TI - Voluntary medication error reporting program in a Japanese national university hospital. AB - BACKGROUND: In Japan, as in other countries, medical accidents arising from human error can seriously damage public confidence in medical services, as well as being intrinsically undesirable. OBJECTIVE: Errors voluntarily reported by the healthcare practitioners in our institution (Kanazawa University Hospital) were considered to assess the contributory factors by using the accumulated error database in the hospital information system. METHODS: Medical errors in our institution during the period from July 1, 2000, to June 30, 2002, were counted using the error reporting system database and were classified. RESULTS: The number of errors reported during the investigation period was 1378, of which 78% were reported by nursing staff. Medication errors involving administration of injectable or oral drugs to inpatients, dispensing, and prescription accounted for about 50% of that number. Among dispensing errors, 53% were detected by patients or their families and 36% by nurses. CONCLUSIONS: The best method of error prevention is to learn from previous errors. For this purpose, the error reporting program is effective. In patient safety management, it is important to take into account the potential risks of future errors, as well as to capture information about errors that have already happened. For safety management, adoption of appropriate information technology (e.g., implementation of a prescription order entry system) is effective in reducing medication errors. However, it is important to note that serious errors can also arise in computer based systems. PMID- 14565815 TI - Fondaparinux for prevention of venous thromboembolism in major orthopedic surgery. AB - OBJECTIVE: To review clinical information related to fondaparinux, a synthetic pentasaccharide recently approved for the prevention of deep-vein thrombosis (DVT) in patients undergoing major orthopedic surgeries and for extended DVT prophylaxis after hip fracture surgery. DATA SOURCES: Primary and review articles were identified by MEDLINE (1983-June 2003) using the key words pentasaccharide, Org31540, SR90107A, DVT prophylaxis, and fondaparinux. Additional sources were found listed in articles, abstracts, and unpublished data on file from the manufacturer. Articles selected were based on their coverage of the pharmacology, pharmacokinetics, safety, and efficacy of fondaparinux. STUDY SELECTION AND DATA EXTRACTION: All of the articles identified were evaluated and all information deemed relevant was included. DATA SYNTHESIS: Fondaparinux is a selective antithrombin-dependent, indirect inhibitor of activated factor Xa. It has a favorable and predictable pharmacokinetic profile when administered subcutaneously, and has a long half-life, allowing once-daily dosing. Fondaparinux lacks in vitro cross-reactivity with heparin-induced antibodies. Major Phase III studies have demonstrated that subcutaneous fondaparinux sodium 2.5 mg given at least 6 hours postoperatively resulted in a 55% reduction in the risk of venous thromboembolism (VTE) in patients undergoing hip fracture surgery, total hip replacement surgery, or knee replacement surgery compared with standard enoxaparin therapy. It has a safety profile similar to that of enoxaparin with respect to clinically relevant major bleeding, including fatal bleeding, nonfatal bleeding, and bleeding requiring repeat surgery. The use of fondaparinux for prolonged prophylaxis after hip fracture has demonstrated further reduction in VTE events without increasing the risk of bleeding. CONCLUSIONS: Fondaparinux is the first of a new class of synthetic factor Xa inhibitors that demonstrated greater efficacy compared with enoxaparin for the prevention of VTE in major orthopedic surgery without an increase in clinically relevant bleeding. Given the favorable cost-effectiveness analysis and improved efficacy profile, fondaparinux should be considered for formulary addition for DVT prophylaxis in patients undergoing hip and knee replacement surgery. In patients undergoing hip fracture surgery, fondaparinux should be considered the DVT prophylaxis of choice. Extended thromboprophylaxis up to 28 days resulted in additional reduction in VTE (both symptomatic and venography-proven DVT) in patients with hip fracture surgery. PMID- 14565817 TI - Comments on the responses of Utteridge et al. (Radiat. Res. 159, 277-278, 2003) to letters about their paper (Radiat. Res. 158, 357-364, 2002). PMID- 14565819 TI - Comments on the paper by Wickliffe et al. (Radiat. Res. 159, 458-464, 2003). PMID- 14565820 TI - Inborn errors in metabolism and 4-boronophenylalanine-fructose-based boron neutron capture therapy. AB - Infusions of boronophenylalanine-fructose complex (BPA-F), at doses up to 900 mg/kg of BPA and 860 mg/kg of fructose, have been used to deliver boron to cancer tissue for boron neutron capture therapy (BNCT). In patients with phenylketonuria (PKU), phenylalanine accumulates, which is harmful in the long run. PKU has been an exclusion criterion for BPA-F-mediated BNCT. Fructose is harmful to individuals with hereditary fructose intolerance (HFI) in amounts currently used in BNCT. The harmful effects are mediated through induction of hypoglycemia and acidosis, which may lead to irreversible organ damage or even death. Consequently, HFI should be added as an exclusion criterion for BNCT if fructose containing solutions are used in boron carriers. Non-HFI subjects may also develop symptoms, such as gastrointestinal pain, if the fructose infusion rate is high. We therefore recommend monitoring of glucose levels and correcting possible hypoglycemia promptly. Except for some populations with extremely low PKU prevalence, HFI and PKU prevalences are similar, approximately 1 or 2 per 20,000. PMID- 14565821 TI - Effects of in vivo exposure to GSM-modulated 900 MHz radiation on mouse peripheral lymphocytes. AB - The aim of this study was to evaluate whether daily whole-body exposure to 900 MHz GSM-modulated radiation could affect spleen lymphocytes. C57BL/6 mice were exposed 2 h/day for 1, 2 or 4 weeks in a TEM cell to an SAR of 1 or 2 W/kg. Untreated and sham-exposed groups were also examined. At the end of the exposure, mice were killed humanely and spleen cells were collected. The number of spleen cells, the percentages of B and T cells, and the distribution of T-cell subpopulations (CD4 and CD8) were not altered by the exposure. T and B cells were also stimulated ex vivo using specific monoclonal antibodies or LPS to induce cell proliferation, cytokine production and expression of activation markers. The results did not show relevant differences in either T or B lymphocytes from mice exposed to an SAR of 1 or 2 W/kg and sham-exposed mice with few exceptions. After 1 week of exposure to 1 or 2 W/kg, an increase in IFN-gamma (Ifng) production was observed that was not evident when the exposure was prolonged to 2 or 4 weeks. This suggests that the immune system might have adapted to RF radiation as it does with other stressing agents. All together, our in vivo data indicate that the T- and B-cell compartments were not substantially affected by exposure to RF radiation and that a clinically relevant effect of RF radiation on the immune system is unlikely to occur. PMID- 14565822 TI - Irradiation increases the interactions of platelets with the endothelium in vivo: analysis by intravital microscopy. AB - Adhesion of platelets to the endothelium is believed to be a major factor contributing to thrombosis and vascular occlusion after radiotherapy or endovascular irradiation. In the present study, platelet-endothelium interactions were analyzed in vivo by intravital microscopy in mesenteric venules of mice according to three parameters: (1) platelet rolling, (2) platelet adhesion, and (3) the presence of platelet clusters. A 10-Gy total-body irradiation of mice resulted in an increase in the frequency of appearance of these three types of platelet-endothelium interactions in postcapillary venules 6 and 24 h after exposure, whereas only minor alterations were seen in large venules. In addition, the duration of platelet adhesion was increased 24 h after irradiation in both postcapillary and large venules. However, P-selectin was not up-regulated on the platelet membrane and platelet-leukocytes were not seen rolling together, suggesting that changes in platelet-endothelial cell interaction result from endothelial cell activation rather than platelet activation. Our data suggest that irradiation transforms resting endothelial cells to a pro-adhesive surface for platelets, which could ultimately lead to thrombosis. PMID- 14565823 TI - Influence of ginger rhizome (Zingiber officinale Rosc) on survival, glutathione and lipid peroxidation in mice after whole-body exposure to gamma radiation. AB - The radioprotective effect of the hydroalcoholic extract of ginger rhizome, Zingiber officinale (ZOE), was studied. Mice were given 10 mg/kg ZOE intraperitoneally once daily for five consecutive days before exposure to 6-12 Gy of gamma radiation and were monitored daily up to 30 days postirradiation for the development of symptoms of radiation sickness and mortality. Pretreatment of mice with ZOE reduced the severity of radiation sickness and the mortality at all doses. The ZOE treatment protected mice from GI syndrome as well as bone marrow syndrome. The dose reduction factor for ZOE was found to be 1.15. The optimum protective dose of 10 mg/kg ZOE was 1/50 of the LD50 (500 mg/kg). Irradiation of the animals resulted in a dose-dependent elevation in the lipid peroxidation and depletion of GSH on day 31 postirradiation; both effects were lessened by pretreatment with ZOE. ZOE also had a dose-dependent antimicrobial activity against Pseudomonas aeruginosa, Salmonella typhimurium, Escherichia coli and Candida albicans. PMID- 14565824 TI - The "pro-drug" RibCys decreases the mutagenicity of high-LET radiation in cultured mammalian cells. AB - We are carrying out studies aimed at reducing the mutagenic effects of high-LET 56Fe ions and 12C ions (56Fe ions, 143 keV/microm; 12C ions, 100 keV/microm) with certain drugs, including RibCys [2-(R,S)-D-ribo-(1',2',3',4'-tetrahydroxybutyl) thiazolidine-4(R)-carboxylic acid]. RibCys, formed by condensation of L-cysteine with D-ribose, is designed so that the sulfhydryl amino acid L-cysteine is released intracellularly through nonenzymatic ring opening and hydrolysis leading to increased levels of glutathione (GSH). RibCys (4 or 10 mM), which was present during irradiation and for a few hours after, significantly decreased the yield of CD59- mutants induced by radiation in AL human-hamster hybrid cells. RibCys did not affect the clonogenic survival of irradiated cells, nor was it mutagenic itself. These results, together with the minimal side effects reported in mice and pigs, indicate that RibCys may be useful, perhaps even when used prophylactically, in reducing the mutation load created by high-LET radiation in astronauts or other exposed individuals. PMID- 14565825 TI - Mitochondrial localization of superoxide dismutase is required for decreasing radiation-induced cellular damage. AB - We investigated the importance of mitochondrial localization of the SOD2 (MnSOD) transgene product for protection of 32D cl 3 hematopoietic cells from radiation induced killing. Four plasmids containing (1) the native human copper/zinc superoxide dismutase (Cu/ZnSOD, SOD1) transgene, (2) the native SOD2 transgene, (3), the SOD2 transgene minus the mitochondrial localization leader sequence (MnSOD-ML), and (4) the SOD2 mitochondrial leader sequence attached to the active portion of the SOD1 transgene (ML-Cu/ZnSOD) were transfected into 32D cl 3 cells and subclonal lines selected by kanamycin resistance. Clonogenic in vitro radiation survival curves derived for each cell clone showed that Cu/ZnSOD- and MnSOD-ML-expressing clones had no increase in cellular radiation resistance (D0=0.89 +/- 0.01 and 1.08 +/- 0.02 Gy, respectively) compared to parent line 32D cl 3 (D0=1.15 +/- 0.11 Gy). In contrast, cell clones expressing either SOD2 or ML Cu/ZnSOD were significantly radioresistant (D0=2.1 +/- 0.1 and 1.97 +/- 0.17 Gy, respectively). Mice injected intraesophageally with SOD2-plasmid/liposome (MnSOD PL) complex demonstrated significantly less esophagitis after 35 Gy compared to control irradiated mice or mice injected intraesophageally with Cu/ZnSOD-PL or MnSOD-ML-PL. Mice injected with intraesophageal ML-Cu/ZnSOD-PL showed significant radioprotection in one experiment. The data demonstrate the importance of mitochondrial localization of SOD in the in vitro and in vivo protection of cells from radiation-induced cellular damage. PMID- 14565826 TI - Bcl2-independent chromatin cleavage is a very early event during induction of apoptosis in mouse thymocytes after treatment with either dexamethasone or ionizing radiation. AB - We have quantified the emergence of early chromatin breaks during the signal transduction phase of apoptosis in mouse thymocytes after treatment with either ionizing radiation or dexamethasone. Dexamethasone at 1 microM can induce significant levels of DNA breaks (equivalent to the amount induced directly by 7.5 Gy ionizing radiation) within 0.5 h of treatment. The execution phase of apoptosis was not observed until 4-6 h after the same treatment. The presence of the Bcl2 transgene under the control of the p56lck promoter almost completely inhibited apoptosis up to 24 h after treatment, but it had virtually no effect on the early chromatin cleavage occurring in the first 6 h. Ionizing radiation induced chromatin cleavage both directly by damaging DNA and indirectly with kinetics similar to the induction of chromatin cleavage by dexamethasone. The presence of the Bcl2 transgene had no effect on the direct or indirect radiation induced cleavage in the first 6 h, but after the first 6 h, the Bcl2 gene inhibited further radiation-induced chromatin cleavage. These results suggest that endonucleases are activated within minutes of treatment with either dexamethasone or ionizing radiation as part of the very early signal transduction phase of apoptosis, and prior to the irreversible commitment to cell death. PMID- 14565827 TI - Effect of Atm disruption on spontaneously arising and radiation-induced deletion mutations in mouse liver. AB - Deletion mutations were efficiently recovered in mouse liver after total-body irradiation with X rays by using a transgenic mouse "gpt-delta" system that harbored a lambda EG10 shuttle vector with the red and gam genes for Spi- (sensitive to P2 lysogen interference) selection. We incorporated this system into homozygous Atm-knockout mice as a model of the radiosensitive hereditary disease ataxia telangiectasia (AT). Lambda phages recovered from the livers of X irradiated mice with the Atm+/+ genotype showed a dose-dependent increase in the Spi- mutant frequency up to sixfold at 50 Gy over the unirradiated control of 2.8x10(-6). The livers from Atm-/- mice yielded a virtually identical dose response curve for X rays with a background fraction of 2.4x10(-6). Structural analyses revealed no significant difference in the proportion of -1 frameshifts and larger deletions between Atm+/+ and Atm-/- mice, although larger deletions prevailed in X-ray-induced Spi- mutants irrespective of Atm status. While a possible defect in DNA repair after irradiation has been strongly indicated in the literature for nondividing cultured cells in vitro from AT patients, the Atm disruption does not significantly affect radiation mutagenesis in the stationary mouse liver in vivo. PMID- 14565828 TI - Quantitative analysis of biological responses to ionizing radiation, including dose, irradiation time, and dose rate. AB - Because biological responses to radiation are complex processes that depend on both irradiation time and total dose, consideration of both dose and dose rate is necessary to predict the risk from long-term irradiations at low dose rates. Here we mathematically and statistically analyzed the quantitative relationships between dose, dose rate and irradiation time using micronucleus formation and inhibition of proliferation of human osteosarcoma cells as indicators of biological response. While the dose-response curves did not change with exposure times of less than 20 h, at a given dose, both biological responses clearly were reduced as exposure time increased to more than 8 days. These responses became dependent on dose rate rather than on total dose when cells were irradiated for 20 to 27 days. Mathematical analysis demonstrates that the relationship between effective dose and dose rate is well described by an exponential function when the logarithm of effective dose is plotted as a function of the logarithm of dose rate. These results suggest that our model, the modified exponential (ME) model, can be applied to predict the risk from exposure to low-dose/low-dose-rate radiation. PMID- 14565829 TI - Radiation tolerance of the rat spinal cord after single and split doses of photons and carbon ions. AB - The sensitivity of the rat spinal cord to single and split doses of radiation and the resulting relative biological effectiveness (RBE) were determined for carbon ion irradiations (12C) in the plateau and Bragg-peak regions. The cranial part of the cervical and thoracic spinal cords of 180 rats were irradiated with one or two fractions of 12C ions or photons, respectively. Dose-response curves for the end point symptomatic myelopathy were established, and the resulting values for the ED50 (dose for 50% complication probability) were used to determine the RBEs. A median latency for myelopathy of 167 days (range, 121-288 days) was found. The ED50 values were 17.1 +/- 0.8 Gy, 24.9 +/- 0.7 Gy (one and two fractions, 12C plateau) and 13.9 +/- 0.8, 15.8 +/- 0.7 Gy (one and two fractions, 12C Bragg peak), respectively. For photons we obtained ED50 values of 24.5 +/- 0.8 Gy for single doses and 34.2 +/- 0.7 Gy when two fractions were applied. The corresponding RBEs were 1.43 +/- 0.08, 1.37 +/- 0.12 (one and two fractions, 12C plateau) and 1.76 +/- 0.05, 2.16 +/- 0.11 (one and two fractions, 12C Bragg peak), respectively. Hematoxylin and eosin staining revealed necrosis of the white matter in the spinal cord in all symptomatic animals. In summary, from one- and two-fraction photon, 12C plateau and Bragg-peak irradiation of the rat spinal cord, we have established RBEs as well as the individual ED50's. From the latter there is a clear indication of repair processes for fractionated photons and 12C plateau ions which are significantly reduced by using Bragg-peak ions. Additional studies are being carried with 6 and 18 fractions to further refine and define the RBE and ED50 values and estimate the alpha/beta ratios. PMID- 14565830 TI - Up-regulation and activation of proteinase-activated receptor 2 in early and delayed radiation injury in the rat intestine: influence of biological activators of proteinase-activated receptor 2. AB - Proteinase-activated receptor 2 (Par2, F2rl1, also designated PAR-2 or PAR2) is prominently expressed in the intestine and has been suggested as a mediator of inflammatory, mitogenic and fibrogenic responses to injury. Mast cell proteinases and pancreatic trypsin, both of which have been shown to affect the intestinal radiation response, are the major biological activators of Par2. Conventional Sprague-Dawley rats, mast cell-deficient rats, and rats in which pancreatic exocrine secretion was blocked pharmacologically by octreotide underwent localized irradiation of a 4-cm loop of small bowel. Radiation injury was assessed 2 weeks after irradiation (early, inflammatory phase) and 26 weeks after irradiation (chronic, fibrotic phase). Par2 expression and activation were assessed by in situ hybridization and immunohistochemistry, using antibodies that distinguished between total (preactivated and activated) Par2 and preactivated Par2. Compared to unirradiated intestine, irradiated intestine exhibited increased Par2 expression, particularly in areas of myofibroblast proliferation and collagen accumulation, after both single-dose and fractionated irradiation. The majority of Par2 expressed in fibrotic areas was activated. Postirradiation Par2 overexpression was greatly attenuated in both mast cell-deficient and octreotide-treated rats. The severity of acute mucosal injury did not affect postirradiation Par2 expression. Mast cells and pancreatic proteinases may exert their fibro-proliferative effects partly through activation of Par2. Par2 may be a potential target for modulating the intestinal radiation response, particularly delayed intestinal wall fibrosis. PMID- 14565831 TI - Comparisons of the skeletal locations of putative plutonium-induced osteosarcomas in humans with those in beagle dogs and with naturally occurring tumors in both species. AB - Osteosarcomas occur from exposures to bone-seeking, alpha-particle-emitting isotopes, particularly plutonium. The skeletal distribution of putative 239Pu induced osteosarcomas reported in Mayak Metallurgical and Radiochemical Plutonium Plant workers is compared with those observed in canine studies, and these are compared with distributions of naturally occurring osteosarcomas in both species. In the Mayak workers, 29% and 71% of the osteosarcomas were in the peripheral and central skeleton, respectively, with the spine having the most tumors (36%). An almost identical distribution of plutonium-induced osteosarcomas was reported for dogs injected with 239Pu as young adults. This distribution of osteosarcomas is quite different from the distributions of naturally occurring osteosarcomas for both species. In the Cooperative Osteosarcoma Study Group in humans (1,736 osteosarcomas from all ages), over 91% of the tumors occurred in the peripheral skeleton. In the Mayo Clinic group of older individuals (>40 years old), over 60% of the osteosarcomas appeared in the peripheral skeleton. The distribution of naturally occurring osteosarcomas in the canine is similar to that in the adult human. The similarities of the distributions of plutonium-associated osteosarcomas in the Mayak workers with those found in experimental studies suggest that many of the reported osteosarcomas may have been associated with plutonium exposures. These results also support the experimental paradigm that plutonium osteosarcomas have a preference for well vascularized cancellous bone sites. These sites have a greater initial deposition of plutonium, but also greater turnover due to elevated bone remodeling rates. PMID- 14565833 TI - Low-dose studies of bystander cell killing with targeted soft X rays. AB - The Gray Cancer Institute ultrasoft X-ray microprobe was used to quantify the bystander response of individual V79 cells exposed to a focused carbon K-shell (278 eV) X-ray beam. The ultrasoft X-ray microprobe is designed to precisely assess the biological response of individual cells irradiated in vitro with a very fine beam of low-energy photons. Characteristic CK X rays are generated by a focused beam of 10 keV electrons striking a graphite target. Circular diffraction gratings (i.e. zone plates) are then employed to focus the X-ray beam into a spot with a radius of 0.25 microm at the sample position. Using this microbeam technology, the correlation between the irradiated cells and their nonirradiated neighbors can be examined critically. The survival response of V79 cells irradiated with a CK X-ray beam was measured in the 0-2-Gy dose range. The response when all cells were irradiated was compared to that obtained when only a single cell was exposed. The cell survival data exhibit a linear-quadratic response when all cells were targeted (with evidence for hypersensitivity at low doses). When only a single cell was targeted within the population, 10% cell killing was measured. In contrast to the binary bystander behavior reported by many other investigations, the effect detected was initially dependent on dose (<200 mGy) and then reached a plateau (>200 mGy). In the low-dose region (<200 mGy), the response after irradiation of a single cell was not significantly different from that when all cells were exposed to radiation. Damaged cells were distributed uniformly over the area of the dish scanned (approximately 25 mm2). However, critical analysis of the distance of the damaged, unirradiated cells from other damaged cells revealed the presence of clusters of damaged cells produced under bystander conditions. PMID- 14565832 TI - Interaction between radiation-induced adaptive response and bystander mutagenesis in mammalian cells. AB - Two conflicting phenomena, the bystander effect and the adaptive response, are important in determining biological responses at low doses of radiation and have the potential to have an impact on the shape of the dose-response relationship. Using the Columbia University charged-particle microbeam and the highly sensitive AL cell mutagenic assay, we reported previously that nonirradiated cells acquired mutagenesis through direct contact with cells whose nuclei had previously been traversed with either a single or 20 alpha particles each. Here we show that pretreatment of cells with a low dose of X rays 4 h before alpha-particle irradiation significantly decreased this bystander mutagenic response. Furthermore, bystander cells showed an increase in sensitivity after a subsequent challenging dose of X rays. Results from the present study address some of the pressing issues regarding both the actual target size and the radiation dose response and can improve on our current understanding of radiation risk assessment. PMID- 14565834 TI - The maximum low-dose RBE of 17.4 and 40 keV monochromatic X rays for the induction of dicentric chromosomes in human peripheral lymphocytes. AB - Schmid et al. recently reported on the maximum low-dose RBE for mammography X rays (29 kV) for the induction of dicentrics in human lymphocytes. To obtain additional information on the RBE for this radiation quality, experiments with monochromatized synchrotron radiation were performed. Monochromatic 17.4 keV X rays were chosen for comparison with the diagnostic mammography X-ray spectrum to evaluate the spectral influence, while monochromatic 40 keV X rays represent a higher-energy reference radiation, within the experiment. The induction of dicentric chromosomes in human lymphocytes from one blood donor irradiated in vitro with 17.4 keV and 40 keV monochromatic X rays resulted in alpha coefficients of (3.44 +/- 0.87) x 10(-2) Gy(-1) and (2.37 +/- 0.93) x 10(-2) Gy( 1), respectively. These biological effects are only about half of the alpha coefficients reported earlier for exposure of blood from the same donor with the broad energy spectra of 29 kV X rays (mean energy of 17.4 keV) and 60 kV X rays (mean energy of 48 keV). A similar behavior is evident in terms of RBEM. Relative to weakly filtered 220 kV X rays, the RBEM for 17.4 and 40 keV monochromatic X rays is 0.86 +/- 0.23 and 0.59 +/- 0.24, respectively, which is in contrast to the RBEM of 1.64 +/- 0.27 for 29 kV X rays and 1.10 +/- 0.19 for 60 kV X rays. It is evident that the monochromatic radiations are less effective in inducing dicentric chromosomes than broad-spectrum X rays with the corresponding mean energy value. Therefore, it can be assumed that, for these X-ray qualities with broad energy spectra, a large fraction of the effects should be attributed predominantly to photons with energies well below the mean energy. PMID- 14565839 TI - Vitamin B(12) deficiency associated with chronic acid suppression therapy. PMID- 14565840 TI - Omeprazole-induced intractable cough. AB - OBJECTIVE: To report a case of chronic, persistent cough induced by omeprazole therapy. CASE SUMMARY: A 42-year-old white woman presented with chronic, persistent cough after omeprazole initiation for treatment of postoperative heartburn. The cough was permanent, dry, and exhausting and worsened at night. Omeprazole therapy was continued for 4 months because the persistent cough was thought to be related to gastroesophageal reflux disease (GERD). However, no cause of persistent, chronic cough was identified. After omeprazole discontinuation, the cough resolved. DISCUSSION: The most common causes of chronic cough in nonsmokers of all ages are postnasal drip syndrome, asthma, and GERD. However, persistent cough without bronchospasm or other pulmonary involvement may occur as a drug adverse effect. According to the US omeprazole package insert, cough is observed as an adverse reaction in 1.1% of patients, although this has not been mentioned in international drug information sources or medical literature. A MEDLINE search (1966-June 2003) using the terms cough, drug related, adverse effects, and omeprazole failed to find any data. In our patient, there was a temporal relationship between cough and medication use, suggesting a causal relationship. An objective causality assessment revealed that the adverse drug reaction was probable. The mechanism is unclear. CONCLUSIONS: Chronic, persistent cough may occur as an adverse effect of omeprazole therapy. Clinicians must be aware of this adverse effect to avoid useless and costly tests. PMID- 14565841 TI - Pharmacist intervention enhances adherence to orlistat therapy. AB - BACKGROUND: Pharmacists, especially those in community practice, should increase their level of intervention in dealing with the nationwide epidemic of obesity since they interact with large numbers of the public on a regular basis. We hypothesized that patients who receive medication for weight loss may have an improved therapeutic outcome if they received additional support from their community pharmacist. OBJECTIVE: To evaluate the impact of pharmacist support on patient persistence with orlistat. METHODS: Pharmacists were trained in basic obesity management skills. Patients who were prescribed orlistat and attending an outpatient nutrition program were invited to participate in the study. All patients agreed to receive pharmaceutical care. Those who lived where the service was available were assigned to the intervention (I) group and those who did not were assigned to the control (C) group. All patients received usual care provided by the outpatient clinic. RESULTS: Thirty patients, 15 in the I group and 15 in the C group, were recruited. Both groups were predominantly women (87%) with a mean +/- SD age of 43.8 +/- 9.7 years. Patients in the I group had significantly greater persistence with orlistat therapy as assessed by duration of therapy (p = 0.006) and number of patients completing the 26-week study (7 I, 2 C; p = 0.046). There was no significant difference in percent of weight loss between groups (p > 0.05). CONCLUSIONS: In this pilot study, patients receiving pharmaceutical care took orlistat longer than the controls and had improved outcome with orlistat therapy. PMID- 14565842 TI - Aripiprazole in an adult with Asperger disorder. AB - OBJECTIVE: To report a case of aripiprazole-responsive adult Asperger disorder. CASE SUMMARY: A 34-year-old white man with lifelong, disabling Asperger disorder and a 20-year history of failed psychotherapeutic and pharmacologic interventions was prescribed aripiprazole, with dramatic symptomatic improvement. DISCUSSION: Multiple prior pharmacologic efforts over several years aimed predominantly at altering serotonin and dopamine neurotransmission resulted in treatment failure due to intolerable adverse effects, exacerbation of underlying symptoms, or nonresponse. Aripiprazole, with a complex profile of neurotransmitter affinities, has produced significant core symptom changes. Improved sociability; increased self-awareness; reduced rigidity, anxiety, and irritability; and reduced preoccupation with circumscribed esoteric interests are among the effects noted. CONCLUSIONS: Previously intractable Asperger disorder symptoms in a 34-year-old man were ameliorated with aripiprazole. PMID- 14565843 TI - Molecular cloning and expression in Escherichia coli of a Trichoderma viride endo beta-(1-->6)-galactanase gene. AB - The nucleotide sequence depicted in Figure 1 has been submitted to the DDBJ nucleotide sequence database under the accession no. AB104898. A gene encoding endo-beta-(1-->6)-galactanase from Trichoderma viride was cloned by reverse transcriptase-PCR and expressed in Escherichia coli. The gene contained an open reading frame consisting of 1437 bp (479 amino acids). The deduced amino acid sequence of the protein showed little similarity with other known glycoside hydrolases. A signal sequence (20 amino acids) was found at the N-terminal region of the protein and the molecular mass of the mature form was calculated to be 50.488 kDa. The gene product expressed in E. coli as a recombinant protein fused with thioredoxin and His(6) tags had almost the same substrate specificity and mode of action as native enzyme purified from a commercial cellulase preparation of T. viride, i.e. recombinant enzyme endo-hydrolysed beta-(1-->6)-galacto oligomers with a DP (degree of polymerization) higher than 3, and it could also hydrolyse alpha-L-arabinofuranosidase-treated arabinogalactan protein from radish. It produced beta-(1-->6)-galacto-oligomers ranging from DP 2 to at least 8 at the initial hydrolysis stage and galactose and beta-(1-->6)-galactobiose as the major products at the final reaction stage. These results indicate that the cloned gene encodes an endo-beta-(1-->6)-galactanase. As far as we know, this is the first time an endo-beta-(1-->6)-galactanase has been cloned. PMID- 14565844 TI - Activator protein-2gamma (AP-2gamma) expression is specifically induced by oestrogens through binding of the oestrogen receptor to a canonical element within the 5'-untranslated region. AB - The activator protein 2 (AP-2) transcription factors are essential proteins for oestrogenic repression of the ERBB2 proto-oncogene in breast cancer cells. In the present study, we have examined the possible oestrogenic regulation of AP-2 genes themselves in breast-tumour-derived lines. As early as 1 h after oestrogen treatment, AP-2gamma mRNA was markedly increased, whereas AP-2alpha was down regulated, but with slower kinetics, and AP-2beta was not affected at all. Addition of anti-oestrogens ablated these effects. Modulation of the protein levels corresponded to changes in the transcript levels, thus suggesting that in oestrogen-treated cells, an inversion of the balance between AP-2alpha and AP 2gamma isoforms occurs. The 5'-untranslated region (5'-UTR) of the human AP 2gamma gene contains one consensus and one degenerate oestrogen-responsive element (ERE). Reporter constructs carrying the AP-2gamma promoter and the 5'-UTR were up-regulated by oestrogens in transient transfection assays. Deletion of the most conserved (but not of the degenerate) ERE from reporter constructs abrogated the oestrogenic response, although both ERE-containing segments were footprinted in DNaseI protection assays. In vitro binding assays demonstrated the ability of oestrogen receptor alpha (ERalpha) to bind to this site, and chromatin immunoprecipitation analysis of the endogenous gene showed that ERalpha occupies this region in response to oestrogens. We conclude that AP-2gamma is a primary oestrogen-responsive gene and suggest that AP-2 proteins may mediate some oestrogenic responses. PMID- 14565845 TI - Ganglioside glycosyltransferases and newly synthesized gangliosides are excluded from detergent-insoluble complexes of Golgi membranes. AB - GEM (glycosphingolipid-enriched microdomains) are specialized detergent-resistant domains of the plasma membrane in which some gangliosides concentrate. Although genesis of GEM is considered to occur in the Golgi complex, where the synthesis of gangliosides also occurs, the issue concerning the incorporation of ganglioside species into GEM is still poorly understood. In this work, using Chinese hamster ovary K1 cell clones with different glycolipid compositions, we compared the behaviour with cold Triton X-100 solubilization of plasma membrane ganglioside species with the same species newly synthesized in Golgi membranes. We also investigated whether three ganglioside glycosyltransferases (a sialyl-, a N-acetylgalactosaminyl- and a galactosyl-transferase) are included or excluded from GEM in Golgi membranes. Our data show that an important fraction of plasma membrane G(M3), and most G(D3) and G(T3), reside in GEM. Immunocytochemical examination of G(D3)-expressing cells showed G(D3) to be distributed as cold detergent-resistant patches in the plasma membrane. These patches did not co localize with a glycosylphosphatidylinositol-anchored protein used as GEM marker, indicating a heterogeneous composition of plasma membrane GEM. In Golgi membranes we were unable to find evidence for GEM localization of either ganglioside glycosyltransferases or newly synthesized gangliosides. Since the same ganglioside species appear in plasma membrane GEM, it was concluded that in vivo nascent G(D3), G(T3) and G(M3) segregate from their synthesizing transferases and then enter GEM. This latter event could have taken place shortly after synthesis in the Golgi cisternae, along the secretory pathway and/or at the cell surface. PMID- 14565846 TI - Prolactin stimulates cell proliferation through a long form of prolactin receptor and K+ channel activation. AB - PRL (prolactin) has been implicated in the proliferation and differentiation of numerous tissues, including the prostate gland. However, the PRL-R (PRL receptor) signal transduction pathway, leading to the stimulation of cell proliferation, remains unclear and has yet to be mapped. The present study was undertaken to develop a clear understanding of the mechanisms involved in this pathway and, in particular, to determine the role of K(+) channels. We used androgen-sensitive prostate cancer (LNCaP) cells whose proliferation is known to be stimulated by PRL. Reverse transcriptase PCR analysis showed that LNCaP cells express a long form of PRL-R, but do not produce its intermediate isoform. Patch-clamp techniques showed that the application of 5 nM PRL increased both the macroscopic K(+) current amplitude and the single K(+)-channel open probability. This single channel activity increase was reduced by the tyrosine kinase inhibitors genistein, herbimycin A and lavandustine A, thereby indicating that tyrosine kinase phosphorylation is required in PRL-induced K(+) channel stimulation. PRL enhances p59( fyn ) phosphorylation by a factor of 2 after a 10 min application in culture. In addition, where an antip59( fyn ) antibody is present in the patch pipette, PRL no longer increases K(+) current amplitude. Furthermore, the PRL stimulated proliferation is inhibited by the K(+) channel inhibitors alpha dendrotoxin and tetraethylammonium. Thus, as K(+) channels are known to be involved in LNCaP cell proliferation, we suggest that K(+) channel modulation by PRL, via p59( fyn ) pathway, is the primary ionic event in PRL signal transduction, triggering cell proliferation. PMID- 14565847 TI - Pancreatic beta-cell granule peptides form heteromolecular complexes which inhibit islet amyloid polypeptide fibril formation. AB - Islet amyloid polypeptide (IAPP), or 'amylin', is co-stored with insulin in secretory granules of pancreatic islet beta-cells. In Type 2 diabetes, IAPP converts into a beta-sheet conformation and oligomerizes to form amyloid fibrils and islet deposits. Granule components, including insulin, inhibit spontaneous IAPP fibril formation in vitro. To determine the mechanism of this inhibition, molecular interactions of insulin with human IAPP (hIAPP), rat IAPP (rIAPP) and other peptides were examined using surface plasmon resonance (BIAcore), CD and transmission electron microscopy (EM). hIAPP and rIAPP complexed with insulin, and this reaction was concentration-dependent. rIAPP and insulin, but not pro insulin, bound to hIAPP. Insulin with a truncated B-chain, to prevent dimerization, also bound hIAPP. In the presence of insulin, hIAPP did not spontaneously develop beta-sheet secondary structure or form fibrils. Insulin interacted with pre-formed IAPP fibrils in a regular repeating pattern, as demonstrated by immunoEM, suggesting that the binding sites for insulin remain exposed in hIAPP fibrils. Since rIAPP and hIAPP form complexes with insulin (and each other), this could explain the lack of amyloid fibrils in transgenic mice expressing hIAPP. It is likely that IAPP fibrillogenesis is inhibited in secretory granules (where the hIAPP concentration is in the millimolar range) by heteromolecular complex formation with insulin. Alterations in the proportions of insulin and IAPP in granules could disrupt the stability of the peptide. The increase in the proportion of unprocessed pro-insulin produced in Type 2 diabetes could be a major factor in destabilization of hIAPP and induction of fibril formation. PMID- 14565849 TI - Eval: a software package for analysis of genome annotations. AB - SUMMARY: Eval is a flexible tool for analyzing the performance of gene annotation systems. It provides summaries and graphical distributions for many descriptive statistics about any set of annotations, regardless of their source. It also compares sets of predictions to standard annotations and to one another. Input is in the standard Gene Transfer Format (GTF). Eval can be run interactively or via the command line, in which case output options include easily parsable tab delimited files. AVAILABILITY: To obtain the module package with documentation, go to http://genes.cse.wustl.edu/ and follow links for Resources, then Software. Please contact brent@cse.wustl.edu PMID- 14565848 TI - Type III secretion proteins PcrV and PcrG from Pseudomonas aeruginosa form a 1:1 complex through high affinity interactions. AB - BACKGROUND: Pseudomonas aeruginosa, an increasingly prevalent opportunistic pathogen, utilizes a type III secretion system for injection of toxins into host cells in order to initiate infection. A crucial component of this system is PcrV, which is essential for cytotoxicity and is found both within the bacterial cytoplasm and localized extracellularly, suggesting that it may play more than one role in Pseudomonas infectivity. LcrV, the homolog of PcrV in Yersinia, has been proposed to participate in effector secretion regulation by interacting with LcrG, which may act as a secretion blocker. Although PcrV also recognizes PcrG within the bacterial cytoplasm, the roles played by the two proteins in type III secretion in Pseudomonas may be different from the ones suggested for their Yersinia counterparts. RESULTS: In this work, we demonstrate by native mass spectrometry that PcrV and PcrG expressed and purified from E. coli form a 1:1 complex in vitro. Circular dichroism results indicate that PcrG is highly unstable in the absence of PcrV; in contrast, both PcrV alone and the PcrV:PcrG complex have high structural integrity. Surface plasmon resonance measurements show that PcrV interacts with PcrG with nanomolar affinity (15.6 nM) and rapid kinetics, an observation which is valid both for the full-length form of PcrG (residues 1-98) as well as a form which lacks the C-terminal 24 residues, which are predicted to have low secondary structure content. CONCLUSIONS: PcrV is a crucial component of the type III secretion system of Pseudomonas, but the way in which it participates in toxin secretion is not understood. Here we have characterized the interaction between PcrV and PcrG in vitro, and shown that PcrG is highly unstable. However, it associates readily with PcrV through a region located within its first 74 amino acids to form a high affinity complex. The fact that PcrV associates and dissociates quickly from an unstable molecule points to the transient nature of a PcrV:PcrG complex. These results are in agreement with analyses from pcrV deletion mutants which suggest that PcrV:PcrG may play a different role in effector secretion than the one described for the LcrV:LcrG complex in Yersinia. PMID- 14565850 TI - Zooprophylaxis or zoopotentiation: the outcome of introducing animals on vector transmission is highly dependent on the mosquito mortality while searching. AB - BACKGROUND: Zooprophylaxis, the diversion of disease carrying insects from humans to animals, may reduce transmission of diseases such as malaria. However, as the number of animals increases, improved availability of blood meals may increase mosquito survival, thereby countering the impact of diverting feeds. METHODS: Computer simulation was used to examine the effects of animals on the transmission of human diseases by mosquitoes. Three scenarios were modelled: (1) endemic transmission, where the animals cannot be infected, eg. malaria; (2) epidemic transmission, where the animals cannot be infected but humans remain susceptible, e.g. malaria; (3) epidemic disease, where both humans and animals can be infected, but develop sterile immunity, eg. Japanese encephalitis B. For each, the passive impact of animals as well as the use of animals as bait to attract mosquitoes to insecticide was examined. The computer programmes are available from the author. A teaching model accompanies this article. RESULTS: For endemic and epidemic malaria with significant searching-associated vector mortality, changing animal numbers and accessibility had little impact. Changing the accessibility of the humans had a much greater effect. For diseases with an animal amplification cycle, the most critical factor was the proximity of the animals to the mosquito breeding sites. CONCLUSION: Estimates of searching associated vector mortality are essential before the effects of changing animal husbandry practices can be predicted. With realistic values of searching associated vector mortality rates, zooprophylaxis may be ineffective. However, use of animals as bait to attract mosquitoes to insecticide is predicted to be a promising strategy. PMID- 14565851 TI - Infection of malaria (Anopheles gambiae s.s.) and filariasis (Culex quinquefasciatus) vectors with the entomopathogenic fungus Metarhizium anisopliae. AB - BACKGROUND: Current intra-domiciliary vector control depends on the application of residual insecticides and/or repellents. Although biological control agents have been developed against aquatic mosquito stages, none are available for adults. Following successful use of an entomopathogenic fungus against tsetse flies (Diptera: Glossinidae) we investigated the potency of this fungus as a biological control agent for adult malaria and filariasis vector mosquitoes. METHODS: In the laboratory, both sexes of Anopheles gambiae sensu stricto and Culex quinquefasciatus were passively contaminated with dry conidia of Metarhizium anisopliae. Pathogenicity of this fungus for An. gambiae was further tested for varying exposure times and different doses of oil-formulated conidia. RESULTS: Comparison of Gompertz survival curves and LT50 values for treated and untreated specimens showed that, for both species, infected mosquitoes died significantly earlier (p < 0.0001) than uninfected control groups. No differences in LT50 values were found for different exposure times (24, 48 hrs or continuous exposure) of An. gambiae to dry conidia. Exposure to oil-formulated conidia (doses ranging from 1.6 x 10(7) to 1.6 x 10(10) conidia/m2) gave LT50 values of 9.69 +/- 1.24 (lowest dose) to 5.89 +/- 0.35 days (highest dose), with infection percentages ranging from 4.4-83.7%. CONCLUSION: Our study marks the first to use an entomopathogenic fungus against adult Afrotropical disease vectors. Given its high pathogenicity for both adult Anopheles and Culex mosquitoes we recommend development of novel targeted indoor application methods for the control of endophagic host-seeking females. PMID- 14565852 TI - Sub-grouping of Plasmodium falciparum 3D7 var genes based on sequence analysis of coding and non-coding regions. AB - BACKGROUND: The variant surface antigen family Plasmodium falciparum erythrocyte membrane protein-1 (PfEMP1) is an important target for protective immunity and is implicated in the pathology of malaria through its ability to adhere to host endothelial receptors. The sequence diversity and organization of the 3D7 PfEMP1 repertoire was investigated on the basis of the complete genome sequence. METHODS: Using two tree-building methods we analysed the coding and non-coding sequences of 3D7 var and rif genes as well as var genes of other parasite strains. RESULTS: var genes can be sub-grouped into three major groups (group A, B and C) and two intermediate groups B/A and B/C representing transitions between the three major groups. The best defined var group, group A, comprises telomeric genes transcribed towards the telomere encoding PfEMP1s with complex domain structures different from the 4-domain type dominant of groups B and C. Two sequences belonging to the var1 and var2 subfamilies formed independent groups. A rif subgroup transcribed towards the centromere was found neighbouring var genes of group A such that the rif and var 5' regions merged. This organization appeared to be unique for the group A var genes CONCLUSION: The grouping of var genes implies that var gene recombination preferentially occurs within var gene groups and it is speculated that the groups reflect a functional diversification evolved to cope with the varying conditions of transmission and host immune response met by the parasite. PMID- 14565853 TI - Giant hydronephrosis mimicking progressive malignancy. AB - BACKGROUND: Cases of giant hydronephroses are rare and usually contain no more than 1-2 litres of fluid in the collecting system. We report a remarkable case of giant hydronephrosis mimicking a progressive malignant abdominal tumour. CASE PRESENTATION: A 78-year-old cachectic woman presented with an enormous abdominal tumour, which, according to the patient, had slowly increased in diameter. Medical history was unremarkable except for a hysterectomy >30 years before. A CT scan revealed a giant cystic tumour filling almost the entire abdominal cavity. It was analysed by two independent radiologists who suspected a tumour originating from the right kidney and additionally a cystic ovarian neoplasm. Subsequently, a diagnostic and therapeutic laparotomy was performed: the tumour presented as a cystic, 35 x 30 x 25 cm expansive structure adhesive to adjacent organs without definite signs of invasive growth. The right renal hilar vessels could finally be identified at its basis. After extirpation another tumourous structure emerged in the pelvis originating from the genital organs and was also resected. The histopathological examination revealed a >15 kg hydronephrotic right kidney, lacking hardly any residual renal cortex parenchyma. The second specimen was identified as an ovary with regressive changes and a large partially calcified cyst. There was no evidence of malignant growth. CONCLUSION: Although both clinical symptoms and the enormous size of the tumour indicated malignant growth, it turned out to be a giant hydronephrosis. Presumably, a chronic obstruction of the distal ureter had caused this extraordinary hydronephrosis. As demonstrated in our case, an accurate diagnosis of giant hydronephrosis remains challenging due to the atrophy of the renal parenchyma associated with chronic obstruction. Therefore, any abdominal cystic mass even in the absence of other evident pathologies should include the differential diagnosis of a possible hydronephrosis. Diagnostic accuracy might be increased by a combination of endourological techniques such as retrograde pyelography and modern imaging modalities. PMID- 14565854 TI - Postural stability is altered by the stimulation of pain but not warm receptors in humans. AB - BACKGROUND: It is now recognized that large diameter myelinated afferents provide the primary source of lower limb proprioceptive information for maintaining an upright standing position. Small diameter afferents transmitting noxious stimuli, however, can also influence motor behaviors. Despite the possible influence of pain on motor behaviors, the effects of pain on the postural control system have not been well documented. METHODS: Two cutaneous heat stimulations (experiment 1: non-noxious 40 degrees C; experiment 2: noxious 45 degrees C) were applied bilaterally on the calves of the subject with two thermal grills to stimulate A delta and C warm receptors and nociceptors in order to examine their effects on postural stability. The non-noxious stimulation induced a gentle sensation of warmth and the noxious stimulation induced a perception of heat pain (visual analogue scores of 0 and 46 mm, respectively). For both experiments, ten healthy young adults were tested with and without heat stimulations of the lower limbs while standing upright on a force platform with eyes open, eyes closed and eyes closed with tendon co-vibration of tibialis anterior and triceps surae muscles. The center of pressure displacements were analyzed to examine how both stimulations affected the regulation of quiet standing and if the effects were exacerbated when vision was removed or ankle proprioception perturbed. RESULTS: The stimulation of the warm receptors (40 degrees C) did not induce any postural deterioration. With pain (45 degrees C), subjects showed a significant increase in standard deviation, range and mean velocity of postural oscillations as well as standard deviation of the center of pressure velocity. The effects of heat pain were exacerbated when subjects had both their eyes closed and ankle tendons vibrated (increased standard deviation of the center of pressure velocity and mean velocity of the center of pressure). CONCLUSIONS: A non-noxious stimulation (40 degrees C) of the small diameter afferents is not a sufficiently intense sensory stimulation to alter the control of posture. A painful stimulation (45 degrees C) of the skin thermoreceptors, however, yielded a deterioration of the postural control system. The observed deteriorating effects of the combined stimulation of nociceptors and Ia afferents (when ankle tendons were vibrated) could result from the convergence of these afferents at the spinal level. This could certainly lead to the hypothesis that individuals suffering from lower limb pain present alterations of the postural control mechanisms; especially populations already at risk of falling (for example, frail elderly) or populations suffering from concomitant lower limb pain and sensory deficits (for example, diabetic polyneuropathy). PMID- 14565856 TI - Interdisciplinary research: putting the methods under the microscope. AB - BACKGROUND: While the desirability of interdisciplinary inquiry has been widely acknowledged, indeed has become 'the mantra of science policy', the methods of interdisciplinary collaboration are opaque to outsiders and generally remain undescribed. DISCUSSION: Many have analysed interdisciplinarity, especially in relation to the creation of new disciplines and institutions. These analyses are briefly outlined. Still, there currently persists a silence about the methods of interdisciplinary collaboration itself, and the core of this paper proposes a template for such methods. SUMMARY: Breaking this silence--by making the methods of interdisciplinary projects transparent--could further invigorate interdisciplinary research. PMID- 14565855 TI - Adenoviral expression of a transforming growth factor-beta1 antisense mRNA is effective in preventing liver fibrosis in bile-duct ligated rats. AB - BACKGROUND: Transforming growth factor-beta (TGF-beta) is a key mediator in establishing liver fibrosis. Therefore, TGF-beta as a causative agent may serve as a primary target for antifibrotic gene therapy approaches. We have previously shown that the adenoviral delivery of a transgene constitutively expressing a TGF beta1 antisense mRNA blocks TGF-beta synthesis in culture-activated hepatic stellate cells and effectively abolishes ongoing fibrogenesis in vitro. METHODS: Ligature of the common bile duct was used to induce liver fibrosis in rats. The effect of the TGF-beta1 antisense on fibrogenesis was analyzed in this model of liver injury. RESULTS: In the present study, we demonstrate that the adenoviral vector directs the synthesis of mRNA quantities that are approximately 8000-fold more abundant than endogenous TGF-beta1 mRNA. In experimentally injured rat livers induced by ligature of the common bile duct, a model for persistent fibrogenesis and cirrhosis, administration of the adenoviral vector abrogates TGF beta-enhanced production of collagen and alpha-smooth muscle actin. Furthermore, the number of cells positive for alpha-smooth muscle actin resulting from active recruitment of activated hepatic stellate cells around the bile ductular structures was significantly reduced in animals after application of Ad5-CMV-AS TGF-beta1. However, the observed elevated serum levels of aspartate aminotransferase, alanine aminotransferase, and bilirubin induced in this obstructive liver injury model were not significantly altered in the presence of the TGF-beta antagonist. CONCLUSION: Taken together, our data provides in vivo evidence that the delivery of TGF-beta1 antisense mRNA specifically abolishes the diverse effects of direct TGF-beta function in ongoing liver fibrogenesis. Therefore, we conclude that the expressed transgene is therapeutically useful for inhibition of TGF-beta effects in diverse applications, ranging from clarification of TGF-beta function in the course of liver injury to the development of novel gene therapeutic approaches. PMID- 14565857 TI - Inhibition of HIV-1 in the central nervous system by IFN-alpha2 delivered by an SV40 vector. AB - In human immunodeficiency virus type 1 (HIV-1)-infected individuals, virus induced production of interferon alpha (IFN-alpha) is impaired. In order to obtain regulated expression of IFN-alpha that responds to HIV-1 infection, a recombinant SV40 vector was designed that carries the human IFN-alpha2 cDNA under the control of the HIV-1 long terminal repeat (LTR) (SV[HIVLTR]IFN). Thus, the IFN-alpha2 gene would be trans-activated on infection with HIV-1. This vector was tested to determine if central nervous system (CNS) cell types that may be potential HIV-1 targets could be transduced and protected from HIV. SV[HIVLTR]IFN transduced NT2 cells, a human neuronal precursor cell line, mature neurons derived from NT2 precursor cells, and human primary monocyte-derived macrophages. IFN-alpha2 expression was retained in mature neurons after SV[HIVLTR]IFN transduced NT2 precursor cells were induced to differentiate using retinoic acid. IFN-alpha expression was detected only after exposing transduced cells to HIV. Furthermore, SV[HIVLTR]IFN-delivered IFN-alpha2 expression significantly inhibited replication of multiple strains of HIV in both NT2 and NT2-derived mature neurons. SV[HIVLTR]IFN transduction also inhibited HIV-1(BaL) replication in human primary monocyte-derived macrophages. Therefore, we have demonstrated the effectiveness of IFN-alpha2, delivered by an SV40 vector driven by HIV-1 LTR as a promoter, to protect several CNS-based, potentially HIV-susceptible cell types. These findings may have implications for therapy of HIV-1 infection in the CNS. PMID- 14565858 TI - Regulation of IL-2 gene expression and nuclear factor-90 translocation in vaccinia virus-infected cells. AB - Nuclear factor-90 (NF-90) has been described as a regulatory subunit of a complex containing DNA-dependent protein kinase (DNA-PK), Ku, and NF-45, which are capable of binding the interleukin-2 (IL-2) enhancer region and stimulating IL-2 gene expression. Vaccinia virus (VV) infection of Jurkat cells induced a nuclear factor that bound specifically to the IL-2 promoter sequence and led to the expression of the IL-2 transcript. Induction of this IL-2 promoter binding factor occurred concomitantly with the induction of NF-90 and translocation of NF-90 to the nucleus. Electrophoretic mobility supershift analysis using specific anti-NF 90 serum suggested the presence of NF-90 in the IL-2 promoter binding complex. As NF-90 can bind to double-stranded RNA (dsRNA) and be phosphorylated by the dsRNA dependent protein kinase, PKR, we investigated whether accumulation of dsRNA in VV-infected cells could regulate IL-2 gene expression. Infection of Jurkat cells with a VV mutant that produces free dsRNA led to similar levels of induced NF-90 within the cell, but the protein remained localized within the cytosol. This mutant did not lead to the accumulation of an IL-2 promoter binding complex or to the synthesis of IL-2 mRNA. Other VV mutants that produced excess dsRNA also inhibited protein binding to the IL-2 enhancer, suggesting that the presence of viral dsRNA has a role in retaining NF-90 in the cytosol and regulating IL-2 gene expression. PMID- 14565859 TI - The role of interferon regulatory factor-1 and interferon regulatory factor-2 in IFN-gamma growth inhibition of human breast carcinoma cell lines. AB - Interferon (IFN) regulatory factor-1 (IRF-1) and IRF-2 play opposing roles in the regulation of many IFN-gamma-inducible genes. To investigate the signal transduction pathway in response to IFN-gamma in light of differences in growth effects, we selected four human breast carcinoma cell lines based on a spectrum of growth inhibition by IFN-gamma. MDA468 growth was markedly inhibited by IFN gamma, and it showed substantial induction of IRF-1 mRNA but little IRF-2 induction. SKBR3 showed little growth inhibition and little induction of IRF-1 mRNA but significant induction of IRF-2 mRNA. HS578T and MDA436 growth inhibition and IRF-1/IRF-2 induction were intermediate. All four cell lines showed intact receptor at the cell surface and Stat1 translocation to the nucleus by immunostaining. By EMSA, there were marked differences in the induced ratio of IRF-1 and IRF-2 binding activity between the cell lines that correlated with growth inhibition. Finally, antisense oligonucleotides specific for IRF-1 attenuated IFN-gamma growth inhibition in MDA436 and MDA468, confirming the direct role of IRF-1 in IFN-gamma growth inhibition. Induction of IRF-1 causes growth inhibition in human breast cancer cell lines, and induction of IRF-2 can oppose this. The relative induction of IRF-1 to IRF-2 is a critical control point in IFN-gamma response. PMID- 14565860 TI - IL-27 and IFN-alpha signal via Stat1 and Stat3 and induce T-Bet and IL-12Rbeta2 in naive T cells. AB - Interleukin-27 (IL-27) supports proliferation of naive CD4(+) T cells and enhances interferon-gamma (IFN-gamma) production by activated T cells and natural killer (NK) cells. We report here that IL-27 induces Stat1 and Stat3 phosphorylation and activation in human and murine cell lines and primary human T cells. IL-27 also induces T-Bet, a Stat1-dependent gene crucial to Th1 cell commitment. Similarly, IFN-alpha activates Stat1 and Stat3 and T-Bet expression in naive T cells. Induction of T-Bet results in upregulation of IL-12Rbeta2 on naive T cells, which is essential for responsiveness to IL-12 and differentiation to a Th1 phenotype. Both IL-27 and IFN-alpha induce expression of IL-12Rbeta2 in T cells. In contrast, IFN-gamma, which activates Stat1 but not Stat3, induces expression of T-Bet but not IL-12Rbeta2 in naive T cells. We propose that IL-27 and IFN-alpha are important for early Th1 commitment and act upstream of IL-12 and IFN-gamma in this pathway. PMID- 14565861 TI - CD8(+)CD28(-) T lymphocytes from HIV-1-infected patients secrete factors that induce endothelial cell proliferation and acquisition of Kaposi's sarcoma cell features. AB - Kaposi's sarcoma (KS) develops more frequently in human immunodeficiency virus type 1 (HIV-1)-infected patients. In this study, we report that molecules released by CD8(+)CD28(-) T lymphocytes from HIV-1-infected patients promote endothelial-cell (EC) growth and induce ECs to acquire spindle cell morphology and upregulation of intercellular adhesion molecule-1 (ICAM-1), E-selectin, and vascular endothelial cell growth factor receptor-3 (VEGFR-3) (a typical feature of the KS cell phenotype). The effects observed on ECs cocultured with in vivo activated CD28(-) cells were partly reproduced when ECs were grown in medium containing interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF alpha). At concentrations similar to those found in the supernatant of in vivo activated CD28(-) cells, the two proinflammatory cytokines sustained EC growth and survival only when combined. We, therefore, conclude that CD28(-) T lymphocytes from HIV-1-infected patients exert their effect on ECs through a mechanism involving both IFN-gamma and TNF-alpha. This finding may have wide implications for our basic understanding of the immunopathology of KS. PMID- 14565862 TI - Microencapsulation of tumor necrosis factor oligomers: a new approach to proinflammatory cytokine inhibition. AB - Antisense oligonucleotides offer great therapeutic potential provided adequate intracellular penetration can be achieved. In this study, we evaluated the effectiveness of microencapsulating antisense oligonucleotides to tumor necrosis factor (TNF) in suppressing TNF release in vitro and in vivo. Microencapsulation of TNF oligomers was performed using albumin to produce microcapsules 0.6-1.0 mum in size that target phagocytic cells. Albumin microcapsules containing fluoresceinated TNF oligomers were incubated with U-937 cells to observe uptake. Microcapsules were added to whole blood and stimulated with Escherichia coli endotoxin. Endotoxin was given intravenously (i.v.) to rats along with 100 mug microencapsulated TNF oligomers to determine TNF inhibition and animal survival. E. coli was given intraperitoneally (i.p.) along with gentamicin and microencapsulated TNF oligomers to assess TNF inhibition and animal survival. The duration of microencapsulated antisense TNF oligomers was also determined in vivo. The results demonstrated rapid uptake of the microcapsules by macrophages after 2 h and 4 h incubation. There was improvement in TNF inhibition in vitro and improved animal survival by microencapsulated antisense in both endotoxin (100% survival) and peritonitis models (70% survival) compared with free antisense oligomers in solution. Microencapsulation extended the duration of action of the oligomers to 72 h. Intracellular targeting of macrophages with antisense oligomers to TNF by microencapsules as a delivery system improves TNF inhibition using the models of whole blood endotoxin stimulation and endotoxic shock and peritonitis in rats. PMID- 14565864 TI - Nrf2 regulates thromboxane synthase gene expression in human lung cells. AB - Thromboxane A(2) synthase (TXAS) converts prostaglandin H(2) to thromboxane A(2), a potent inducer of vaso-constriction and platelet aggregation. TXAS expression level is cell type preferential; high in hematopoietic cells and low in nonhematopoietic cells. We previously showed that p45 NF-E2 activated the TXAS promoter in hematopoietic cells via binding to the nucleotides -86/-77 from the transcriptional start site [Yaekashiwa and Wang (2002) J. Biol. Chem. 277, 22497 22508]. We reported here that, by transient transfection analysis, this region was also critical for TXAS trans-activation in the A549 and WI-38 lung cells. Mutation of the NF-E2 site greatly reduced TXAS promoter activity in these two types of cells. Using stably transfected A549 cells, we showed that an NF-E2 mutation retained only 0.25% of the wild-type promoter activity. Ecotopic expression of NF-E2 related factors showed that Nrf2, but not Nrf1, Nrf3, or Bach1, activated TXAS promoter in a dose-dependent manner. Furthermore, chromatin immunoprecipitation assay using the stably transfected A549 cells demonstrated that Nrf2 bound the TXAS NF-E2 site in vivo. TXAS gene thus utilizes the same cis acting element but different trans-acting factors to confer cell-preferential expression. We also showed that forced expression of p300 upregulated TXAS gene in a dose-dependent manner. Mutation of NF-E2 site, but not TATA or initiator site, abolished the p300-mediated activation of TXAS gene. PMID- 14565865 TI - MZF6D, a novel KRAB zinc-finger gene expressed exclusively in meiotic male germ cells. AB - Spermatogenesis takes place in the seminiferous tubule in the testes and culminates in the production of spermatozoa (male gametes). Here we report the identification of a novel mouse zinc-finger gene, MZF6D, which is selectively expressed in meiotic spermatocytes. The MZF6D protein contains an N-terminally located repressor domain, a KRAB domain, followed by at least seven successive Kruppel zinc-finger motifs. The KRAB domain of MZF6D, which consists of a KRAB A box and the newly identified KRAB C box, has previously been shown to interact with TIF1beta, which is the common corepressor of all KRAB zinc-finger proteins. Northern blot analysis shows that the expression of MZF6D is restricted to testes. This was confirmed by RT-PCR analysis of a panel of mouse tissues. In situ hybridization of sections from adult mouse testes localizes the expression to meiotic spermatocytes, suggesting a specific role for MZF6D in the regulation of spermatogenesis. PMID- 14565866 TI - Expression of a conserved mouse stress-modulating gene, Bre: comparison with the human ortholog. AB - Mouse Bre, an evolutionarily conserved stress-modulating gene, like its human counterpart, is expressed in multiple alternative transcripts. The main transcript, which is ubiquitously expressed, encodes a protein that binds tumor necrosis factor receptor 1 (TNF-R1) and downregulates TNF-induced activation of NF-kappaB. Alternative splicing of mouse Bre occurs only at the 5' region of the gene, generating either nonfunctional transcripts or transcripts that can encode putative protein isoforms differ at the N-terminal sequence. In contrast, alternative splicing of human BRE occurs at either or both ends of the gene; only the 3' alternative splicing can generate functional transcripts that encode putative protein isoforms differ at the C-terminus, occurrence of the 5' alternative splicing only results in forming nonfunctional transcripts. Unlike the human BRE alternative transcripts which are coexpressed at considerable levels with the main transcript, the mouse counterparts are expressed in a restricted pattern and generally in low abundance except in the heart. Both species, however, share a type of Bre alternative transcripts generated by cryptic splicing at a nonstandard, noncanonical acceptor site. Thus, a highly conserved gene in two species can generate alternative transcripts different in both of the sequence structure and expression pattern, as well as a similar class of transcripts resulting from unconventional transcript processing. PMID- 14565867 TI - Promoter activity and gene structure of rabbit FKBP52. AB - A 0.9-kb fragment situated directly upstream of the first ATG of rabbit FKBP52 is rich in acceptor sites for transcription factors, contains a CAAT box at -197 and could represent the proximal promoter of this immunophilin. Transvection analysis of this fragment showed strong promoter activity on the expression of a reporter gene. Deletions at the 5' end of this fragment showed that a basic sequence of 155 base pairs upstream of the CAAT box was sufficient to enhance luciferase expression an average 220-fold compared to the empty vector. This sequence, which contains acceptor sites for transcription factors of the EGR family and heat shock factors, is closely homologous to 110 base pairs situated directly 5' of FKBP52 exon 1 in human chromosome 12p13.3, suggesting that these transcription factors could be involved in the regulation of the gene in both species. Furthermore, the upstream region of RbFKBP52 contains a large proportion of SINEs (C-repeats, Alu analogs), some of which include the A and B boxes required for transcription of RNA polymerase III, and poly A tracts. RbFKBP52, like HuFKBP52, is made up of 10 exons and 9 introns, a feature shared with other large immunophilins such as FKBP65 and Cyclophilin 40, and which appears widely conserved. PMID- 14565868 TI - Clustering of the B cell receptor is not required for the apoptotic response. AB - We examined the role of BCR cell membrane redistribution in anti-IgM-induced apoptosis in three human B cell lines, RA#1, 2G6, and MC116, that differ in their relative levels of sIgM expression. The apoptotic response was found to be dependent on the nature of the anti-IgM and the cell line. In the cell lines, RA#1 and MC116, sIgM aggregated into patches that were insensitive to the disruption of cholesterol-rich membrane microdomains by nystatin or beta-MCD. The B cell line 2G6 was able to reorganize sIgM into a tight coalescent cap upon anti IgM treatment. However, in this case, the lipid raft inhibitors nystatin and beta MCD disrupted the patching. In 2G6 cells, BCR-mediated apoptosis was not affected by nystatin treatment, whereas it increased in beta-MCD pretreated cells. Thus, no evident correlation was found between apoptosis and BCR cell membrane redistribution or lipid raft formation in either of the three cell lines. The data indicate that the apoptotic signal transduction pathway is independent of BCR translocation into lipid rafts and/or aggregation. PMID- 14565869 TI - Kinetics of the apoptotic response induced by anti-IgM engagement of the B cell receptor is dependent on the density of cell surface immunoglobulin M expression. AB - The effect of B cell receptor (BCR) density on anti-BCR-induced apoptosis was assessed in Ramos cell lines, expressing low, medium, or high levels of surface IgM (sIgM(LO), sIgM(MED), sIgM(HI)). All cells required a 6-mug/ml threshold of anti-IgM to elicit apoptosis. Anti-IgM treatment of sIgM(LO) cells induced growth inhibition and limited dose-independent apoptosis. Anti-IgM treatment of sIgM(MED) cells induced dose-independent death with a 32-h lag. Ligation of the BCR in the sIgM(HI) cells induced rapid apoptosis beginning by 6 h, which was dose-dependent. Secondary crosslinking reagents did not affect apoptosis, and this effect was independent of anti-IgM concentration, time, or sIgM density. These results suggest that the response to BCR engagement strongly depends on the cell surface receptor density. PMID- 14565870 TI - Expression of the proto-oncogene Axl in renal cell carcinoma. AB - In this investigation, we examined the role of the Axl proto-oncogene in renal cell carcinoma (RCC). Axl is a tyrosine kinase receptor implicated in myeloid leukogenesis, and has been found to be overexpressed in lung cancers and breast cancers. Axl has been described to act as a mitogenic factor along with its ligand Gas-6. Axl has also shown to have a role in apoptosis, cell adhesion, and chemotaxis. The differential expression of the Axl RNA transcript was examined in 20 pairs of matched normal kidney and clear cell RCC patient samples. We found that there was a significant increase in the steady-state levels of Axl mRNA in the RCC compared with the normal kidney pair (Student's paired t-test P < 0.001). There was also a significant increase in Axl expression overall in RCC compared to normal kidney (P < 0.03). Western blotting was utilized to determine Axl protein levels in six out of the 20 pairs of the normal/RCC matched pairs. Overall, the level of expression was not significantly different between the paired normal kidneys and kidney tumors, but the detected Axl protein appeared to be at slightly different molecular weights. Primers were constructed for the two known Axl variant, RT-PCR performed, but no differences were observed in the expression of each variant. Next, we performed a gene silencing experiment utilizing double-stranded RNA constructed to silence the Axl gene in the 293 transformed kidney cell line. There was a 50% decrease in Axl gene expression in the RNAi transfected over control cells. In addition, flow cytometry performed to determine DNA content showed a 30% increase in G1/G0 cells, which were transfected with axl RNAi compared to control. Altogether, these findings suggest an overexpression of Axl as part of a proliferative phenotype in RCC. PMID- 14565871 TI - Endoscopy and endourology coding update for 2003. PMID- 14565873 TI - Measurement of renal anatomy for prediction of lower-pole caliceal stone clearance: reproducibility of different parameters. AB - BACKGROUND AND PURPOSE: The architecture of the lower renal pole seems to have a considerable influence on caliceal stone clearance after different therapeutic modalities. The published data are partially inconsistent, and publications on reproducibility are completely lacking. The aim of this study was to evaluate the intraobserver and interobserver reproducibility of different measures of lower calix anatomy. MATERIALS AND METHODS: We studied the intraobserver and interobserver reproducibility of parameters describing the lower-pole anatomy that are significant for treatment outcome. Forty renal units without urologic disease were analyzed by five independent urologists. Infundibular length (IL), infundibular width (IW), and lower infundibulopelvic angles (LIP) were measured by the Elbahnasy (LIP I), Keeley (LIP II), and Gupta (LIP III and LIP IV) methods. Statistical analysis of each parameter and investigator was performed. RESULTS: All LIP angles showed low interobserver correspondence: correlation coefficients (CC) did not exceed 0.44 (P < 0.05). Even the relatively clearly defined parameters IW and IL achieved CCs of only 0.63 and 0.49, respectively. The intraobserver correlation achieved better results: 0.73 (LIP I), 0.84 (LIP II), 0.73 (LIP III), 0.65 (LIP IV), 0.88 (IL), and 0.82 (IW). With the Elbahnasy method, almost all renal units were classified as favorable for stone persistence after shockwave lithotripsy. With the Keeley and Gupta methods, more than 50% of the kidneys were defined as having low clearance probability. CONCLUSIONS: Assessment of the chosen parameters is difficult and shows high interobserver variation. Inexperience in measuring the specific angles and low imaging quality can limit correct evaluation. The large number of kidneys with anatomy inappropriate for clearance of lower-pole stones may explain the poor outcome of shockwave treatment for stones in a lower calix. Prospective studies will determine the clinical value of anatomic assessments. PMID- 14565872 TI - Cavitation bubble cluster activity in the breakage of kidney stones by lithotripter shockwaves. AB - BACKGROUND AND PURPOSE: There is strong evidence that cavitation bubble activity contributes to stone breakage and that shockwave-bubble interactions are involved in the tissue trauma associated with shockwave lithotripsy. Cavitation control may thus be a way to improve lithotripsy. MATERIALS AND METHODS: High-speed photography was used to analyze cavitation bubble activity at the surface of artificial and natural kidney stones during exposure to lithotripter shockwaves in vitro. RESULTS: Numerous individual bubbles formed on the surfaces of stones, but these bubbles did not remain independent but rather combined to form clusters. Bubble clusters formed at the proximal and distal ends and at the sides of stones. Each cluster collapsed to a narrow point of impact. Collapse of the proximal cluster eroded the leading face of the stone, and the collapse of clusters at the sides of stones appeared to contribute to the growth of cracks. Collapse of the distal cluster caused minimal damage. CONCLUSION: Cavitation mediated damage to stones is attributable, not to the action of solitary bubbles, but to the growth and collapse of bubble clusters. PMID- 14565874 TI - Update on minimally invasive management of ureteral strictures. AB - PURPOSE: To review the role of minimally invasive management in ureteral stricture disease. MATERIALS AND METHODS: A literature search was performed on the MEDLINE database through 2002 concerning endoscopic treatment of patients with ureteral strictures. RESULTS: Many endourologic methods are available for ureteral strictures. Ureteral dilation may be accomplished in most cases, with various rates of success depending on stricture etiology, location, and length. Endoureterotomy also leads to long-term patency in properly selected cases and appears to be superior to dilation alone. CONCLUSIONS: Significant advances in technique and technology have improved our ability to treat ureteral strictures without the need for open surgery in most patients. PMID- 14565875 TI - Video-assisted minilaparotomy in urology. AB - Minimally invasive surgery has gained wide acceptance as a method of reducing postoperative pain and curtailing the convalescence period. We have devised a modified surgical technique of video-assisted surgery through minilaparotomy (VAMS). This technique is a hybrid of conventional open and laparoscopic surgery that combines the benefits of both techniques by reducing postoperative pain and scarring, as in laparoscopy, but at the same time maintaining the safety of conventional open surgery. Video-assisted procedures have become standard as a result of our experience with 245 consecutive patients operated on between January 1993 and January 2001. The VAMS is a minimally invasive technique that is safe, feasible, standardized, and reproducible with a short learning curve. This technique can be an alternative to open and laparoscopic surgery in daily urologic practice. PMID- 14565877 TI - Laparoscopic radical excision of urachal sinus. AB - Persistent urachus is a rare congenital anomaly. Various types of remnants have been described including cyst, alternating sinus, patent urachus, diverticulum, and sinus. The most common presenting symptom of urachal sinus is umbilical discharge. Radical excision of the remnant, with or without a bladder cuff, is essential to prevent future malignant degeneration or recurrence of the remnant. Although open surgical excision has been the treatment of choice for many years, the laparoscopic approach has become an attractive alternative because of its association with less postoperative pain, better cosmesis, and rapid convalescence. Laparoscopic radical excision of a urachal sinus was performed in a 16-year-old female patient who presented with umbilical discharge. PMID- 14565878 TI - Simple model for training in the laparoscopic vesicourethral running anastomosis. AB - PURPOSE: To create a simple model for training in the complex technique of laparoscopic vesicourethral anastomosis after radical prostatectomy. MATERIALS AND METHODS: A model simulating the performance of a laparoscopic running urethrovesical anastomosis was fashioned using skin of a supermarket chicken set in a laparoscopic training box. A circular continuous anastomosis was performed, following exactly the technique we use in vivo for performing a vesicourethral anastomosis after laparoscopic radical prostatectomy. Details regarding the use of the right or the left needle holder, needle positioning, precise stitch position, and use of forehand or backhand stitch were strictly respected in order to reproduce the continuous stitch currently done in vivo in our department. The learning curve was analyzed in terms of the time necessary to perform the task and the quality of the anastomosis. RESULTS: The model was created and used by the junior authors during their year of fellowship in advanced laparoscopic urology. The authors had no previous experience with hands-on laparoscopic suturing but have assisted in a great number of laparoscopic radical prostatectomies. The time required for performing the anastomosis on the model declined from 75 minutes initially to 20 minutes after the trainees had performed 20 anastomoses each. After this training, it was possible to create a watertight running laparoscopic vesicourethral anastomosis in patients in a mean time of 40 minutes (range 30-55 minutes). CONCLUSION: This simple model allows the trainee in laparoscopic urology to acquire the skills necessary to perform a laparoscopic vesicourethral anastomosis, one of the most complex steps in laparoscopic radical prostatectomy, as well as to develop dexterity and facility in laparoscopic manipulation of needles, sutures, and fragile tissues. PMID- 14565876 TI - Laparoscopic upper-pole heminephrectomy for ectopic ureter: surgical technique. AB - A duplex kidney associated with a poorly functioning upper-pole segment is commonly associated with incontinence, voiding dysfunction, and urinary tract infections. A standard treatment option for this condition is upper-pole heminephrectomy. With the continued development of minimally invasive urology, this technique can now be safely performed laparoscopically. This report details step by step our technique of laparoscopic upper-pole heminephrectomy. Key points include placement of a catheter in the normal ureter at the start of the case, full mobilization of the upper-pole ureter away from the renal hilum, and precise identification of the vasculature supplying the upper pole. Laparoscopic upper pole heminephrectomy for ectopic ureter is safe and reproducible and offers the patient the typical postoperative benefits of laparoscopic surgery. PMID- 14565879 TI - Laser-assisted fluoroscopic puncture: a new technique for accessing the kidney. AB - PURPOSE: We developed a laser pointer device that is mounted on the image intensifier of a C-arm fluoroscope. The laser-assisted fluoroscopic puncture technique (LAFPT) is an alternative method of accessing the kidney. MATERIALS AND METHODS: The LAFPT was assessed both in an in vitro model and in a series of cases. The in vitro model was composed of Foley balloons filled with contrast medium of different volumes (2-6 mL) embedded in silicone pads. The total fluoroscopy times and attempts required for a successful puncture with LAFPT were compared with those of conventional fluoroscopic access. RESULTS: The number of attempts required to puncture the 2-mL balloons was significantly greater with the conventional fluoroscopic technique than with LAFPT. The fluoroscopy time in vitro was reduced 50% with LAFPT. However, there was no statistically significant difference in the time required for the two techniques in the clinical series. CONCLUSION: The laser-assisted fluoroscopic puncture technique allows accurate positioning of a needle with reduced radiation exposure. PMID- 14565880 TI - Symptomatic port-site hernia associated with a non-bladed trocar after laparoscopic live-donor nephrectomy. AB - We describe what appears to be the first case of a port-site hernia at the site of insertion of a 10-mm non-bladed trocar, which was discovered 4 days after laparoscopic live-donor nephrectomy. We now close all 10-mm port sites in adults and all 5-mm port sites in children; this can be done easily using the Carter Thomason device. PMID- 14565881 TI - Baskets in the kidney: an old problem in a new situation. AB - A potential complication of using a zero-tip basket to remove stones from a lower lower pole calix of the kidney is impaction in this position. We report an endourologic technique involving a hydrogel-coated ureteral dilator to facilitate closure of the wires and removal of the basket following lithotripsy to fragment the stone. PMID- 14565882 TI - Ureteral catheterization controversies. PMID- 14565883 TI - Management of lower-pole caliceal stones. AB - A series of 205 urologists answered questions about their choice of treatment for lower-caliceal stones. The preferred approaches were extracorporeal shockwave lithotripsy (SWL) for stones <1 cm and percutaneous nephrolithotomy (PCNL) for those >2 cm. For stones of 1 to 2 cm, 65% preferred SWL and 30% would advise PCNL. Thus, SWL is recommended for lower-caliceal stones more frequently than is justified by published success rates. Continued efforts need to be made to inform practicing urologists regarding the most appropriate therapy for patients with lower-pole stones >1 cm. PMID- 14565884 TI - Transurethral resection of prostrate and suprapubic ballistic vesicolithotrity for benign prostatic hyperplasia with vesical calculi. AB - Bladder calculi associated with benign prostatic hyperplasia are fairly common, and a popular treatment is transurethral resection of the prostate (TURP) with endoscopic lithotripsy as a combined procedure. We have been routinely using a Reuter's suprapubic trocar and cannula for continuous irrigation during TURP for medium-size and large glands. Rather than performing the vesicolithotripsy per urethra, we have found it easier and faster to use the suprapubic route, especially because the suprapubic tract was already partly made by the Reuter's trocar and half sheath. We describe the technique in detail and explain why we find it easier to extract the stone by the suprapubic rather than the urethral route. PMID- 14565885 TI - Per-urethral endoscopic management of bladder stones: does size matter? AB - Large urinary bladder stones occupying the whole lumen are still encountered. Conventionally, an open suprapubic cystolithotomy (SPCL) has been the accepted treatment of choice. The other method described is suprapubic percutaneous lithotripsy. The present paper describes complete stone clearance in adults by the urethral route using a conventional nephroscope, a pneumatic lithotripter, and an evacuator. An additional procedure to treat outlet obstruction is performed when required. Although the procedure takes a long time, stone clearance is complete, and the procedure is free from any complications. This technique should be offered to all adults, as it has negligible morbidity. PMID- 14565886 TI - Externally readjustable device to regulate sling tension in stress urinary incontinence: preliminary results. AB - BACKGROUND AND PURPOSE: Slings, especially those called "tension free," have low complication rates and good long-term success rates. However, they still have problems with either an excess or a lack of tension, which produces voiding difficulties or urinary leakage persistence, respectively. The effectiveness of a new adjustable sling for the treatment of stress urinary incontinence (SUI) has been evaluated. PATIENTS AND METHODS: Between November 1999 and May 2002, 113 consecutive patients were operated for SUI by means of a new adjustable sling. The results were analyzed retrospectively. The average follow-up time was 22 months (range 6-36 months). RESULTS: Objective success was achieved in 108 of 113 patients (95.5%), but subjectively, only 90.3% of the patients were completely satisfied with the procedure because of persistent urge incontinence in 6 cases (5.3%). Morbidity was minimal. CONCLUSIONS: This new minimally invasive treatment allows readjustment of sling tension at the immediate or mid-term postoperative period. It is applicable to primary and recurrent SUI and has shown encouraging results. PMID- 14565887 TI - Swarming of Proteus mirabilis over ureteral stents: a comparative assessment. AB - BACKGROUND AND PURPOSE: Encrustation on indwelling ureteral stents is commonly related to the presence of urease-producing bacteria that elevate the pH of the urine through the hydrolysis of urea, resulting in the precipitation of calcium and magnesium salts. Using a model previously shown to measure accurately the ability of Proteus mirabilis to swarm over catheter surfaces (Eur J Clin Microbiol Infect Dis 1999;18:206), we investigated the ability of this organism to swarm over three ureteral stents with potential encrustation-resistance properties. MATERIALS AND METHODS: Three commercially available ureteral stents were selected for evaluation: a low surface-energy stent, a hydrogel-coated stent, and a silicone stent. Ten-microliter aliquots of a 4-hour culture of P. mirabilis 296 in Trypticase soya (TSA) broth was inoculated 5 mm from a 1-cm channel cut out from TSA plates. Ten-millimeter stent sections were placed as bridges across the central channel adjacent to the inocula. Time to pathogen crossing was measured. RESULTS: The mean time (+/- SD) to pathogen migration across the three test materials was 15.9 +/- 6.1, 19.8 +/- 9.5, and 29.7 +/- 14.3 hours for the low surface-energy, hydrogel-coated, and silicone stents, respectively. Statistical analysis revealed significant differences between the crossing times of the low surface-energy (P = 0.001) and hydrogel-coated (P = 0.034) stents compared with silicone but not between the low surface-energy and hydrogel-coated stents (P = 0.387). CONCLUSION: Migration of P. mirabilis 296 across silicone stents was significantly reduced compared with low surface-energy and hydrogel-coated stents. These findings suggest that P. mirabilis may have a lower affinity for silicone stents, which may translate into a reduced risk of infection with P. mirabilis and associated stent encrustation. PMID- 14565888 TI - Comment on "Supracostal approach in percutaneous nephrolithotomy: experience of 102 cases". PMID- 14565889 TI - Selective processing of negative emotional information in children and adolescents with Generalized Anxiety Disorder. AB - OBJECTIVES: Research investigating attentional bias for emotional information using the modified Stroop task in younger anxious populations has produced equivocal results. The present data investigated the replicability in younger participants of the prototypical adult finding of Mathews and MacLeod (1985) with patients with Generalized Anxiety Disorder (GAD). METHOD: A sample of 19 child and adolescent patients with GAD and 19 controls completed the modified Stroop paradigm with threat, depression-related, positive and neutral words. RESULTS: The data revealed a selective Stroop interference effect for negative emotional information in the GAD patients, relative to the performance of the controls. CONCLUSIONS: The results provide evidence of a modified Stroop effect for negative emotional material in children and adolescents with GAD, suggesting that modified Stroop processing in younger generally anxious populations broadly mirrors the profile of results in adults. PMID- 14565890 TI - Dissociation as a mediator of the relationship between recalled parenting and the clinical correlates of auditory hallucinations. AB - BACKGROUND: This study of patients who experience auditory hallucinations examined the potential role of dissociation as a mediator in the relationship between memories of parental behaviours during childhood and psychopathology in adulthood. METHOD: Thirty-six patients who experienced auditory hallucinations completed standardized measures of recalled parental behaviours, beliefs about voices, depression and dissociation. Regression analyses were used to test for mediating effects. RESULTS: The findings support a model where dissociation mediates the relationship between paternal overprotection and depression. There were no significant findings regarding the relationship between recalled parenting and beliefs about voices. DISCUSSION: The clinical and research implications of the study are outlined, emphasizing dissociation as a potential treatment target. PMID- 14565891 TI - The effects of the psychiatric label 'borderline personality disorder' on nursing staff's perceptions and causal attributions for challenging behaviours. AB - OBJECTIVES: The aim of the study was to investigate how the psychiatric label 'borderline personality disorder' (BPD) affected staff's perceptions and causal attributions about patients' behaviour. METHODS AND DESIGN: The study utilized a within-participants questionnaire methodology and participants comprised qualified mental health nursing staff. The questionnaire contained descriptions of challenging behaviour in which the patient was described with a diagnosis of depression, schizophrenia or BPD. Participants were asked to identify a likely cause of the behaviour and then on a Likert-type scale rate attributions of internality, stability, globality and controllability. In addition they recorded their level of sympathy with the patient and their optimism for change. RESULTS: Patients with a label of BPD attracted more negative responses from staff than those with a label of schizophrenia or depression. Causes of their negative behaviour were rated as more stable and they were thought to be more in control of the causes of the behaviour and the behaviour itself. Staff reported less sympathy and optimism towards patients with a diagnosis of BPD and rated their personal experiences as more negative than their experiences of working with patients with a diagnosis of depression or schizophrenia. CONCLUSIONS: Staff regard patients with a BPD label to be more in control of negative behaviour than patients with a label of schizophrenia or depression. In accord with Weiner's (1985) model, attributions of control were inversely related to staff sympathy. Addressing attributions of control may provide a means to modify staff sympathy towards patients with a diagnosis of BPD and counteract their negative experiences. PMID- 14565892 TI - Pure and guided self-help for full and sub-threshold bulimia nervosa and binge eating disorder. AB - OBJECTIVE: The study compared the efficacy of self-help without and with guidance (referred to as 'pure' and 'guided' self-help), using a cognitive behavioral self help manual (Fairburn, 1995) for binge eating. DESIGN AND METHODS: A sample of 31 participants with bulimia nervosa, subthreshold bulimia nervosa and binge-eating disorder were assigned randomly to one of the self-help levels for 16 weeks, after four weeks of baseline observations. RESULTS: The results indicated that both forms of self-help treatments had a modestly positive and sustained effect on the participants' eating problems. Intention-to-treat analyses showed that participants reduced their mean number of objective bulimic episodes and purging behaviour by 33% and 17% over the course of the treatment. The corresponding reduction levels for the treatment completers (N = 18) were 58% and 61%, respectively. Furthermore, there were no significant differences between the pure and guided self-help groups in terms of outcome, reflecting the probable insignificance of guidance for the broader group of individuals with binge-eating problems. At follow-up, no further significant improvement or deterioration was observed in the ensuing six months compared with the post-treatment data. CONCLUSION: Given the heterogeneity of the diagnostic groups in the present study, resulting in high external validity, and the conservative nature of the analyses, self-help is discussed as a viable means of initial treatment for binge eating. PMID- 14565893 TI - An exploration of pre-sleep cognitive activity in insomnia: imagery and verbal thought. AB - OVERVIEW: Patients with insomnia commonly report that unpleasant intrusive thoughts and images prevent them falling asleep. Previous research has documented the topics people with insomnia think about during the pre-sleep period, but has not yet distinguished between different types of cognitive activity. Given that research relating to the anxiety disorders suggests that different types of cognitive activity, in particular images and verbal worry, are functionally independent, the present study aimed to provide a detailed phenomenological investigation of imagery and verbal thought in insomnia. DESIGN AND METHOD: A semi-structured interview, designed to assess the content and management of pre sleep images and verbal thought, was administered to individuals with sleep-onset insomnia (N = 34) and good sleepers (N = 38) immediately following an afternoon nap. During the nap, heart rate and sleep-onset latency were measured objectively. RESULTS: The insomnia group was more likely than the good sleeper group to report negative images than positive images. The start, direction and stopping of pre-sleep images was rated as less controllable than pre-sleep verbal thoughts. CONCLUSION: Participants disengaged more from images compared with verbal thoughts, and the insomnia patients reported feeling less calm and relaxed at the end of the salient verbal thought compared with the good sleepers. PMID- 14565894 TI - Horizontal rhythmical eye movements consistently diminish the arousal provoked by auditory stimuli. AB - OBJECTIVES: Theoretical models implicating the orienting reflex as an explanatory mechanism in the eye-movement desensitization and reprocessing (EMDR) treatment protocol are contrasted and tested empirically. We also test whether EMDR effects are due to a distraction effect. DESIGN: A repeated measure design is used in two experiments. The first experiment employed two independent variables, eye condition (moving vs. stationary) and tone (a pseudo-randomized series of low and high intensity tones). In Expt 2, eye condition was replaced by attentional demand conditions (low or high). In both cases, electrodermal responses served as the dependent variable. METHOD: Participants were recruited from the Psychology Department at Cardiff University. In Expt 1, participants were required to either pursue a moving stimulus following auditory challenge or engage in an eyes stationary task. In Expt 2, the task following auditory challenge required participants to identify specific items from letter strings in low and high attentional demand conditions. RESULTS: Lower levels of electrodermal arousal were identified in tasks eliciting eye movements, compared to no eye movements. This effect was not due to the attentional requirements of the task. CONCLUSIONS: Eye movements following auditory challenge result in an effect of psychophysiological de-arousal. This supports the reassurance reflex model of EMDR proposed by MacCulloch and Feldman (1996). PMID- 14565895 TI - Feigned intellectual deficits on the Wechsler Adult Intelligence Scale-Revised. AB - OBJECTIVES: To examine whether individuals instructed to fake intellectual impairment on the WAIS-R employed common faking tactics and to assess the accuracy of detection of faking tactics employed. METHOD: Two groups of participants were instructed to fake 'mental handicap' on the WAIS-R. The first group included incarcerated young offenders and the second group included postgraduate students and research fellows from a university psychology department. RESULTS: Both groups produced faked mean IQ scores within the 'mental retardation' range. There were, however, unusual features in the answers of both groups, including unusual error types. The faking tactics employed were readily detected. CONCLUSIONS: The results suggest that an individual's WAIS-R performance should be interpreted with due consideration given to both the quantitative and the qualitative aspects of their presentation. PMID- 14565896 TI - How robust is performance on the National Adult Reading Test following traumatic brain injury? AB - OBJECTIVE: To investigate whether National Adult Reading Test (NART) performance may be impaired by severe traumatic brain injury (TBI). METHOD: A sample of 26 people who had been given a NART within 12 months of a severe TBI was given a second NART at least 12 months after the first NART. RESULTS: Mean performance on the second NART was significantly better than performance on the first NART. Of the participants, 11 (42%) showed an improvement of more than 5 IQ points in respect of the verbal IQ estimates based on their NART scores, with three participants showing an improvement of 20 points. In applying the NART to determine the presence of an acquired intellectual impairment, use of the first NART scores alone would have resulted in such impairments being missed in at least 25% of a subsample of 16 participants for whom an actual verbal IQ had been obtained at the time of the first NART. CONCLUSION: An NART given within 12 months of a severe TBI runs the risk of significantly underestimating pre-morbid IQ. If applied in this context, it is recommended that the NART score is used in conjunction with other methods of estimation, such as those based on demographic data. PMID- 14565897 TI - Functional specificity of memory function associated with different subregions of the medial temporal lobe. PMID- 14565898 TI - Pleconaril for infantile enterovirus meningitis. PMID- 14565899 TI - Bacterial meningitis: diagnosis and treatment. AB - Bacterial meningitis, once almost universally fatal, remains a cause of serious neurologic illness and subsequent disability. The initial diagnosis, although sometimes clear-cut, can be subtle and difficult. Appropriate intervention may have a favorable influence on outcome. The advent of corticosteroids and brain imaging has improved management but created controversy concerning the sequence at which various interventions should be introduced. Empiric treatment with antibiotics is important in the initial stages. Complications can arise, such as seizures, increased intracranial pressure, and ischemia. The prognosis remains unfavorable for some affected individuals in spite of advances in antibiotic management and supportive care. PMID- 14565900 TI - Whipple disease. AB - Whipple disease is a rare disease caused by infection with the bacterium Tropheryma whippelii. Humans are the only known host for the infection. The signs of systemic infection include gastrointestinal problems, weight loss, and arthritis. Signs of central nervous system infection include cognitive changes, supranuclear gaze palsy, altered level of consciousness, and movement disorders. The diagnosis is based on clinical findings as well as microscopic examination of biopsy specimens and, more recently, polymerase chain reaction (PCR) analysis, which has high sensitivity and specificity. Although the organism historically has been difficult to culture, several recent attempts have been successful. Antibiotic treatment is recommended for 1 year while monitoring the clinical signs and cerebrospinal fluid PCR results. PMID- 14565902 TI - Memantine in moderate to severe Alzheimer's disease. PMID- 14565901 TI - Bioterrorism and the nervous system. AB - Recent events of war, terrorist attacks, and mail-borne anthrax exposure have produced increasing awareness of potential bioterrorism attacks in the United States and other parts of the world. Physicians and healthcare personnel play a key role in identifying potential bioterrorist attacks. Early recognition and preparedness for bioterrorism-associated illnesses is especially important for neurologists because most bioterrorism agents can directly or indirectly affect the nervous system. This article reviews the neurologic manifestations, diagnosis, and treatments of syndromes caused by potential bioterrorism agents, as well as the potential side effects of vaccines against some of these agents. PMID- 14565903 TI - Executive dyscontrol in normal aging: normative data, factor structure, and clinical correlates. AB - Although there is ample evidence of frontal system dysfunction in old age, the clinical significance of these impairments has not been well studied. In this article, we examine the factor structure of putative executive measures in a sample of well elderly subjects. Three stable factors emerged, but only one was associated with measures of functional status. This factor was most strongly associated with relatively simple bedside executive measures that might prove useful in dementia screening. In contrast, a second factor, dominated by the Wisconsin Card Sorting Task, was weakly associated with disability. These data suggest that certain executive measures may be more relevant to functional outcomes, and hence dementia case finding, than others. Moreover, associations with functional status should not be casually extrapolated from regional frontal clinical correlations, particularly with the dorsolateral prefrontal cortex. PMID- 14565904 TI - Face memory and its disorders. AB - Face recognition is an essential biologic and social skill. Accurate recognition depends on the ability to encode, store, and retrieve distinct memory representations for the faces of countless individuals encountered in everyday life. In addition, face memory records must be integrated with specific biographic and name information in order to allow the recognition of each person's unique identity. Converging evidence from functional imaging, cortical electrical recording, and neuropsychologic studies suggests that face memory operations in the human brain are mediated by a distributed neural system. Components of this network include specialized memory storage sites within temporal neocortex that interact with medial temporal lobe and prefrontal cortical areas during face memory encoding and retrieval. Selective damage to these neuroanatomic regions gives rise to face recognition disorders characterized by memory loss or memory distortion. PMID- 14565905 TI - Amusia and musicogenic epilepsy. AB - Amusia and musicogenic epilepsy are clinical disorders that provide a window into the localization of music in the brain. Classic clinical studies of patients with these disorders, coupled with more recent studies employing modern neuroimaging and sophisticated neuropsychologic paradigms, have converged in helping to elucidate the complex neural systems that are utilized in decoding music. The notion of cerebral dominance for music has been replaced by a concept of modular but interconnected networks that have wide bilateral localization in the brain and that are molded both by genetics and experience. These disorders also provide insight into the important interface between music and emotion. PMID- 14565906 TI - Visual agnosia. AB - The visual agnosias are an intriguing class of clinical phenomena that have important implications for current theories of high-level vision. Visual agnosia is defined as impaired object recognition that cannot be attributed to visual loss, language impairment, or a general mental decline. At least in some instances, agnostic patients generate an adequate internal representation of the stimulus but fail to recognize it. In this review, we begin by describing the classic works related to the visual agnosias, followed by a description of the major clinical variants and their occurrence in degenerative disorders. In keeping with the theme of this issue, we then discuss recent contributions to this domain. Finally, we present evidence from functional imaging studies to support the clinical distinction between the various types of visual agnosias. PMID- 14565907 TI - Intravascular cooling for fever control. PMID- 14565908 TI - Neuromonitoring: brain oxygenation and microdialysis. AB - Patients with cerebral lesions run a high risk of developing cerebral hypoxic and ischemic damage due to secondary insults. To minimize the risk of secondary cerebral hypoxia and ischemia, new monitoring techniques of cerebral oxygenation and metabolism have been developed and may help to understand the pathophysiology of secondary brain damage for a better treatment and outcome in critical patients. Cerebral microdialysis is a relatively new technique for measuring brain molecules of the extracellular space. The technical aspects, the interpretation of the commonly measured parameters, the use of the two commonly used oxygenation parameters (jugular venous oxygen saturation and monitoring of brain tissue PO(2) and the microdialysis technique to monitor cerebral metabolism in patients with head injury), subarachnoid hemorrhage, and ischemic stroke are considered. Pitfalls of the techniques and their future potential are discussed. PMID- 14565910 TI - [Cytokines value as a sepsis and mortality predictor in elderly patients with fever]. AB - BACKGROUND AND OBJECTIVE: Proinflammatory cytokines (IL-1beta, IL-6, TNF-alpha) are excellent predictive factors of tissue damage, inflammation and infection. However, there is not sufficient data about their usefulness in elderly patients with acute septic pathology. Our objective was to identify the cytokines related to bacteremia and those that predicted a bad prognosis in elderly patients. PATIENTS AND METHOD: Prospective study carried out during 1999. Patients aged >= 60 years with temperature >= 38 C admitted to the emergency ward. We determined IL-1beta, IL-6, TNF-alpha and C-reactive protein (CRP); cultures were done according to the infectious source. On the 4th day, cytokines and CRP were recorded again. The follow-up was completed until cure or death. RESULTS: 50 patients were included (29 males). Median age was 75.6 (SD: 8.98). The etiology was infectious in 44 (88%): respiratory in 29 (66%), urinary in 8 (18%) and other sources in 7. Thirteen patients had bacteremia (32%): Escherichia coli (4), Streptococcus pneumoniae (5) and others (4). Ten patients died (20%). Median values on admission were CRP : 6.67 mg/dl (NV 0.8), TNF-alpha: 29 pg/ml (NV 0 20), IL-1beta: 7 pg/ml (NV 15) and IL-6: 121 pg/ml (NV 5). 4th day values were: 4.23 mg/dl, 22 pg/ml, 1 pg/ml and 41 pg/ml, respectively. The levels of IL-1b in the 2nd determination were significantly lower in females (p = 0.019). Initial TNF-alpha (p = 0.033), IL-1beta (p = 0.013) and IL-6 (p = 0.031) values were significantly higher in bacteremia patients. IL-6 values on the 4th day were higher in patients who died (p = 0.06). In patients who died, IL-6 levels were higher in the 2nd determination (p = 0.09). CONCLUSIONS: Median values of all cytokines were higher in the bacteremia population. Patients who died showed higher levels of IL-6 on the 4th day. PMID- 14565909 TI - Hypertonic saline for cerebral edema. AB - Raised intracranial pressure (ICP) is a major contributor to the mortality of many conditions encountered in a neurologic intensive care unit. Achieving a sustained reduction in ICP in patients with intracranial hypertension remains a challenge. Treatment with hyperosmolar agents is one of the few options that are available, and mannitol is currently the most commonly used agent. However, hypertonic saline solutions have recently emerged as a potentially safer and more efficacious alternative to mannitol. PMID- 14565911 TI - [A validation study of the Spanish versions of the Clinical Anxiety Scale and the Physician Questionnaire for the assessment of anxiety disorders]. AB - BACKGROUND AND OBJECTIVE: To validate the Spanish versions of both the Clinical Anxiety Scale (CAS) and the Physician Questionnaire (PQ) to assess the severity of symptoms in patients with Anxiety Disorders (AD) treated in ambulatory psychiatric care facilities. PATIENTS AND METHOD: Observational, prospective and multi-centre study (15 psychiatric facilities included) with a follow-up of 1 to 6 weeks in clinically stable or unstable AD patients. We assessed their convergent validity (with the Hamilton Anxiety Rating Scale as reference), discriminant validity (as compared with the Clinical Global Impression), reliability (internal consistency, test-retest and inter-raters) and sensitivity to change. RESULTS: One hundred and sixty one AD patients were included (48 stable and 133 unstable patients). Both the CAS and the PQ showed appropriate convergent (r >= 0.70) and discriminant validity (p 0.001), internal consistency (Cronbach's a > 0.75), test-retest reliability (intraclass correlation coefficient [ICC] > 0,90), inter-raters reliability (ICC > 0.90) and sensitivity to change (p < 0.0001; effect size >= 1.5). CONCLUSIONS: The Spanish versions of both rating scales show appropriate validity and reliability and thus can be used to assess the severity of anxiety symptoms and their change in AD patients treated in ambulatory care. PMID- 14565912 TI - [Effectiveness of Spanish tuberculosis control programmes]. AB - BACKGROUND AND OBJECTIVE: An assessment of tuberculosis programmes was carried out in Spain in 1996. The present article looks at trends in tuberculosis control since then. MATERIAL AND METHOD: A questionnaire asking about indicators, referred to year 2000, was sent to Spain's 17 autonomous regions, as well as to the autonomous cities of Ceuta and Melilla. Results were compared with those obtained in 1996. RESULTS: Improvements, i.e. implementation of more activities, were observed in 77.8% of operative programmes. However, 16.7% of regions have no program yet, while in 27.8% of them, control activities remain scarce. Castilla Leon, Catalunya, Ceuta, Madrid, Murcia, Valencia and Galicia were the regions exhibiting more control activities. CONCLUSIONS: Although the trend is positive, many programmes still have a limited effectiveness. PMID- 14565913 TI - [Indicators of sepsis severity]. PMID- 14565914 TI - [B-type ventricular peptide]. PMID- 14565916 TI - [Clinical comments on the new classification of diffuse interstitial lung diseases]. PMID- 14565915 TI - [A critical review of the pragmatic clinical trial]. AB - Schwartz and Lellouch proposed in 1967 to differentiate between pragmatic and explanatory clinical trials. The pragmatic approach has been understood in different ways and has also been the object of some criticism. Yet different authors have asked to perform pragmatic clinical trials, especially in issues requiring a therapeutical decision. In order to clarify the present situation of the pragmatic trial, a bibliographic review has been performed. Between 1976 and 2002, 95 original articles on clinical trials conducted following a pragmatic design have been found. Only four followed strictly the approach developed by Schwartz et al. In a pragmatic clinical trial, it is usually not possible to blind the interventions and for this reason randomization is imperfect. This limitation casts doubts about the validity of the frequentistic methods in the intention-to-treat evaluation of the results of a pragmatic trial. This experimental approach demands an evaluation according to the decision-making theory. Following the Bayes theorem, the credibility and the previous probability of a hypothesis conditions its posterior probability. It has been agreed that Bayesian statistics is a suitable instrument for the evaluation of a pragmatic clinical trial, but the lack of adequate informatics' programs has limited seriously its application. Recently,some new programs (WinBUGS, TreeAge) have been developed and applied to the decision analysis in some primary care therapeutic questions. It seems possible to predict that, thanks to the new informatics'programs on Bayesian statistics, the pragmatic clinical trial will experience, in the short term, an important revival. PMID- 14565917 TI - [Plasmapheresis? It probably means plasma exchange]. PMID- 14565918 TI - [Hepatocellular carcinoma producing syndrome of inappropriate antidiuretic hormone secretion]. PMID- 14565919 TI - [Type A aortic intramural hemorrhage and Horner syndrome]. PMID- 14565920 TI - [Strongyloides stercoralis superinfection more than 10 years after leaving endemic area]. PMID- 14565921 TI - [Different approach in high-cardiovascular-risk women, compared to men]. PMID- 14565922 TI - [Adverse reactions to drugs among hospitalized elderly patients]. PMID- 14565925 TI - Dominant-negative effect of the c-fos family gene products on inducible NO synthase expression in macrophages. AB - Activation of murine peritoneal macrophages or the macrophage cell line RAW264 with IFN-gamma and bacterial lipopolysaccharide promotes a transient up regulation of c-fos family gene expression following inducible NO synthase (iNOS) production. Since introduction of a double mutation into the two AP-1-binding sites in the iNOS promoter region reduced the promoter activity to 25% of the authentic one in activated RAW264 cells, the induced c-Fos/AP-1 may promote iNOS expression in activated macrophages. Surprisingly, overexpression of c-fos in activated macrophages completely suppressed the production of iNOS, but not that of IL-6 and IL-1beta. The regulatory effect was also observed by overexpression of c-fos, c-jun or fosB on the promoter activity as deduced from transfection experiments. However, the mutation of AP-1-binding sites in the promoter region did not abrogate the regulatory effect of c-fos and the effect of c-fos was diminished by co-transfection with c-jun, but not with fosB, suggesting no relation between the regulatory effect and a c-Fos/AP-1 complex. Expression of NF IL6 (C/EBPbeta), whose gene product can make a non-functional heterodimer with c Fos family proteins, was transiently induced in activated macrophages. Overexpression of NF-IL6 in activated RAW264 cells augmented iNOS promoter activity and reduced the regulatory effect of c-fos overexpression. Thus, overproduction of c-Fos family proteins acts as a dominant-negative-type regulator on iNOS expression in activated macrophages. PMID- 14565926 TI - IL-5-deficient mice are susceptible to experimental autoimmune encephalomyelitis. AB - Experimental autoimmune encephalomyelitis (EAE) is an animal model commonly used to investigate mechanisms involved in the activation of self-reactive T cells. Whereas auto-reactive T(h)1 cells are believed to be involved in the generation of EAE, T(h)2 cells can induce EAE in immunocompromised hosts. Since the T(h)2 cytokine IL-5 can influence the nature and severity of disease, we investigated the role of IL-5 in the EAE model. Wild-type C57BL/6J and IL-5(-/-) mice were immunized with myelin oligodendrocyte glycoprotein (MOG)(35-55) peptide and the development of EAE observed. Our results show that IL-5(-/-) mice developed EAE with a similar day of onset and comparable severity to wild-type mice. Primed T cells isolated from IL-5(-/-) mice proliferated equally to wild-type cells in response to antigen challenge with MOG(35-55). Antigen-specific T cells from IL 5(-/-) mice produced IFN-gamma and tumor necrosis factor-alpha, but no IL-4 or IL 10, indicating that a predominant T(h)1 environment was induced following immunization. No differences in the types of cells infiltrating into the central nervous system were observed between IL-5(-/-) and wild-type mice. Our results suggest that IL-5 is not directly involved in the initiation or effector phase of MOG(35-55)-induced EAE in immunocompetent C57BL/6J mice. PMID- 14565927 TI - MHC class II loading of high or low affinity peptides directed by Ii/peptide fusion constructs: implications for T cell activation. AB - CD4(+) T cells recognize peptides presented on the cell surface of antigen presenting cells in the MHC class II context. The biosynthesis and transport of MHC class II molecules depend on the type II transmembrane invariant chain (Ii) and are tightly regulated processes. Ii is known to bind to the MHC class II peptide-binding groove via its class II-associated Ii peptide (CLIP) region early in the biosynthetic pathway to prevent premature peptide binding. In this study we have genetically exchanged CLIP with peptides of either high or low affinity for the class II peptide binding groove and utilized the properties of Ii to manipulate MHC class II loading. An inducible promoter controlled expression of the Ii/peptide fusion constructs, and presentation at different expression levels was studied. Both peptides were excised from Ii and presented on MHC class II molecules as shown by liquid chromatography-tandem mass spectrometry, but the high affinity peptide was presented more efficiently than the low affinity peptide. Both peptides were efficient in eliciting T cell responses at high Ii/peptide concentration independent of the duration of T cell stimulation. The peptides were also able to elicit an IL-2 response at low expression levels; however, the kinetic differed as the T cells required longer duration of T cell contact to reach a significant T cell response. This probably reflects the number of class II/peptide complexes at the cell surface and is discussed. PMID- 14565928 TI - Recognition mechanism of non-peptide antigens by human gammadelta T cells. AB - The majority of gammadelta T cells in adult human blood exhibit Vgamma2/Vdelta2 TCR and specifically respond to various kinds of non-peptide antigens. In this study, we comparatively analyzed the CDR3 repertoires of Vgamma2-gamma and Vdelta2-delta chain genes in the adult and cord blood. It was confirmed that the vast majority of adult gammadelta T cells exhibited Vgamma2-gamma chains bearing a Jgamma1.2 segment with no or short N-region and Vdelta2-delta chains with a conserved hydrophobic residue (leucine, valine or isoleucine) at position 97 encoded by N-region of Vdelta/Jdelta junction (deltaL97). The cord blood cells stimulated with pyrophosphomonoester antigen in vitro showed preferential expansion of the gammadelta T cells expressing Vgamma2- and Vdelta2-TCR chains with these structural features as compared with those stimulated with a polyclonal mitogen phytohemagglutinin. TCR gene transfer studies indicated that alanine substitution of lysine at position 108 in Jgamma1.2 (gammaK108) or deltaL97 abrogated the responsiveness of Vgamma2/Vdelta2-TCR to all kinds of the non-peptide antigens without affecting the response to anti-CD3 antibody. Furthermore, alanine substitution of arginine at position 51 in Vdelta2 segment (deltaR51) adjacent to gammaK108 in the Vgamma2/Vdelta2-gammadelta TCR also abolished the antigen responsiveness. These results strongly suggested that a hydrophobic and two cationic residues (deltaL97, gammaK108 and deltaR51) clustered in a particular topology at the surface edge of the pocket structure of Vgamma2/Vdelta2-gammadelta TCR played essential roles in the recognition of non peptide antigens. PMID- 14565930 TI - Thymus leukemia antigen (TL)-specific cytotoxic T lymphocytes recognize the alpha1/alpha2 domain of TL free from antigenic peptides. AB - The thymus leukemia antigens (TL) belong to the MHC class Ib family and can be recognized by CD8-dependent or -independent cytotoxic T lymphocytes (CTL) showing TL, but not H-2, restriction. We previously reported that the CTL epitope is TAP independent and in the present study we further characterize the recognition mechanism of CD8-dependent TL-specific TCRalphabeta CTL. We first prepared empty TL tetramers by way of peptide-independent folding with recombinant proteins produced in an Escherichia coli expression system, and showed that TL-specific CTL recognized TL without putative TL-associated peptide and/or post translational modifications of TL by mammalian and insect cells. We next prepared transfectants expressing various chimeric TL molecules with mouse or human MHC class I as well as chimeric TL tetramers with recombinant proteins produced by insect cells, and demonstrated that chimeric TL whose alpha3 domain was replaced by that of H-2K(b), but not of HLA-A2, was sufficient for binding and activation of TL-specific CTL. These results indicate that TL-specific CTL recognize predominantly their alpha1/alpha2 domain as an epitope(s) and that the binding activity to the murine CD8 of the alpha3 domain of H-2K(b) is sufficient to induce their CTL activity, although it is known to be weaker than that of TL, but stronger than that of HLA. The results taken together indicate that CD8-dependent TL-specific TCRalphabeta CTL recognize an epitope(s) of the alpha1/alpha2 domain of TL free from antigenic molecules, and that CD8 plays an important role in stable interactions between TL and their corresponding TCR. PMID- 14565929 TI - Differential activation of signal transducer and activator of transcription (STAT)3 and STAT5 and induction of suppressors of cytokine signalling in T(h)1 and T(h)2 cells. AB - Cytokines direct the differentiation of naive CD4(+) T cells into either IFN gamma-producing T(h)1 cells or IL-4-producing T(h)2 cells. In this study, we analyzed the activation of signal transducer and activator of transcription (STAT)1, STAT3 and STAT5 (together with STAT4 and STAT6), and the expression of the recently identified suppressor of cytokine signalling (SOCS) proteins, in differentiated T(h)1 and T(h)2 cells, both before and after re-stimulation with anti-CD3 and anti-CD28. In addition to the polarized activation of STAT4 in T(h)1 cells and STAT6 in T(h)2 cells, we found that STAT3 and STAT5 are selectively activated in T(h)1 cells after differentiation. This activation of STAT3 and STAT5 was maintained after TCR re-stimulation. The selective activation of STAT3 and STAT5 in T(h)1 cells was associated with differential induction of SOCS molecules. After re-stimulation, SOCS1 expression was significantly increased in T(h)2 cells, but not in T(h)1 and non-polarized 'T(h)' cells. Additionally, the level of CIS was higher in T(h)2 cells compared with T(h)1 and T(h) cells. In contrast, the expression of SOCS3 was higher in T(h)1 cells. The differential induction of SOCS proteins was paralleled by the differential expression of cytokines in re-stimulated T(h)1 and T(h)2 cells (IFN-gamma and IL-4/IL-13 respectively). Our results suggests that STAT3 and STAT5, possibly regulated by the SOCS proteins, may play a role in the differentiation of T(h) cells, and in the maintenance of the T(h)1 and T(h)2 phenotype. PMID- 14565931 TI - Combined deficiency in CD44 and Fas leads to exacerbation of lymphoproliferative and autoimmune disease. AB - Patients with mutations in Fas develop autoimmune lymphoproliferative disease (ALPS), while their family members with similar mutations are often normal, thereby suggesting that additional factors may play a role in the development of ALPS. In the current study, we tested the role of CD44 in the development of lymphoproliferative disease by generating CD44(-/-)/Fas(-/-) mice, which failed to express CD44 and Fas, and compared them to CD44(+/+)/Fas(-/-) mice that expressed CD44, but not Fas. The results showed that CD44(-/-)/Fas(-/-) mice developed a more severe lymphoproliferative and autoimmune disease when compared to CD44(+/+)/Fas(-/-) mice. This was indicated by increased numbers of cells in their lymph nodes, and a greater proportion of B220(+)CD4(-)CD8(-) (double negative) T cells as well as antibodies against single-stranded DNA and chromatin. The heightened severity of lymphoproliferative disease seen in CD44(-/ )/Fas(-/-) mice correlated with increased resistance of T cells, but not B cells, to undergo activation-induced cell death (AICD). The current study suggests that deficiency in CD44 in combination with a defect in one of the molecules involved in the death receptor family such as Fas can further down-regulate AICD, and exacerbate the lymphoproliferative and autoimmune disease. PMID- 14565932 TI - Differential role for IL-7 in inducing lung Kruppel-like factor (Kruppel-like factor 2) expression by naive versus activated T cells. AB - Kruppel-like Factor 2 [KLF2, also called lung Kruppel-like factor (LKLF)] is a transcription factor shown to be necessary for the maintenance of naive T cells. KLF2 is expressed in both naive and memory cells, and is proposed to promote quiescence in these populations. During T cell stimulation, both KLF2 protein and mRNA are down-regulated, and loss of KLF2 appears to be critical for full T cell activation. It is unclear, however, how KLF2 expression is maintained in naive T cells. Recently it was proposed that IL-7, which is known to promote KLF2 re expression in antigen-stimulated T cells, may also induce KLF2 expression in naive T cells. Here we address this issue by comparing the impact of IL-7 on KLF2 expression in naive and activated T cells. Use of bcl-2 transgenic T cells allowed us to uncouple the requirements for IL-7 in preserving naive T cell survival from its role in maintaining KLF2 expression. Our data demonstrates that IL-7 signals are not required for KLF2 maintenance in naive T cells, suggesting that this cytokine has distinct effects on KLF2 expression in naive versus activated T cells. PMID- 14565933 TI - Differential contribution of the immunoreceptor tyrosine-based inhibitory motifs of human leukocyte-associated Ig-like receptor-1 to inhibitory function and phosphatase recruitment. AB - Leukocyte-associated Ig-like receptor (LAIR)-1 is an inhibitory receptor expressed on most human leukocytes. It contains two immunoreceptor tyrosine-based inhibitory motifs (ITIM) in its cytoplasmic tail and recruits phosphatases upon phosphorylation. Here we show that both ITIM are required for full inhibition of cellular responses and optimal phosphatase recruitment. Mutation of the C terminal ITIM still allows partial inhibition of the cytotoxic activity of the NK like YT.2C2 cells, while mutation of the N-terminal ITIM completely abolishes this inhibitory activity. In contrast, in rat basophilic leukemia (RBL) cells, both mutants of LAIR-1 are partially effective. This is reflected in phosphorylation of these mutants in the different cell types upon pervanadate treatment. However, in both YT.2C2 cells and RBL cells, only the mutant containing the N-terminal ITIM recruits Src homology 2 domain-containing tyrosine phosphatase-2 (SHP-2), while the mutant containing the C-terminal ITIM does not. In RBL cells the mutant containing only the N-terminal ITIM also binds SHP-1, although to a lesser extent than wild-type LAIR-1. We find that in Jurkat T cells Lck is required for the association of SHP-1 with LAIR-1. Co-expression with Lck in 293T cells leads to phosphorylation of both wild-type LAIR-1 and the mutant containing only the N-terminal ITIM, while the mutant lacking this ITIM is not phosphorylated. These results indicate that Lck, or another Src family kinase, is essential for the consecutive phosphorylation of the N- and C-terminal ITIM. Our data imply that the N-terminal ITIM is dominant in LAIR-1 signaling, but that both ITIM contribute to an optimal inhibitory function. PMID- 14565934 TI - TCR-mediated hyper-responsiveness of autoimmune Galphai2(-/-) mice is an intrinsic naive CD4(+) T cell disorder selective for the Galphai2 subunit. AB - Heterotrimeric Gi signaling regulates immune homeostasis, since autoimmunity occurs upon disruption of this pathway. However, the role of the lymphocyte expressed Galphai subunits (Galphai2 and 3) on T cell activation and cytokine production is poorly understood. To examine this role, we studied T lymphocytes from mice deficient in the Galphai2 or Galphai3 subunits. Galphai2(-/-) but not Galphai3(-/-) splenocytes were hyper-responsive for IFN-gamma and IL-4 production following activation through the TCR. Galphai2(-/-) T cells had a relaxed costimulatory requirement for IL-2 secretion and proliferation compared to wild type cells. Purified naive Galphai2(-/-) T cells produced more IL-2 than naive wild-type T cells following TCR activation, indicating that the hyper-responsive cytokine profile was not due to the expanded Galphai2(-/-) memory T cells, but involved an intrinsic T cell alteration. Cytokine hyper-responsiveness was not seen when purified Galphai2(-/-) T cells were stimulated with phorbol myristic acetate/ionomycin, localizing the alteration to a proximal TCR-specific signaling pathway. Galphai2(-/-) CD4(+) T cells were distinguished from wild-type or Galphai3(-/-) T cells by a globally augmented TCR-induced calcium response. These findings indicate that Galphai2(-/-) mice have an intrinsic CD4(+) T cell abnormality in TCR signaling which may be one cause of augmented T cell effector function and Galphai2(-/-) autoimmune susceptibility. PMID- 14565935 TI - Rapamycin antagonizes cyclosporin A- and tacrolimus (FK506)-mediated augmentation of linker for activation of T cell expression in T cells. AB - The discovery of new immunosuppressive drugs such as rapamycin, cyclosporin A (CsA) and tacrolimus (FK506) has been very useful for preventing graft rejection and autoimmune disease. However, these drugs are not specific, and are associated with side-effects and toxicities. Therefore, understanding the molecular mechanisms of these drugs is important for designing specific immunosuppressants. Here, we show that in contrast to CsA and FK506, rapamycin blocks activation induced expression of the linker for activation of T cells (LAT), a signaling molecule critical for initiating TCR signaling. Thus, whereas CsA and FK506 strongly enhanced TCR- and phorbol myristate acetate-induced LAT expression in T cells, rapamycin effectively inhibited activation-induced LAT expression. Importantly, these opposite effects were mutually antagonistic, as rapamycin acted as a potent antagonist for both CsA and FK506. Because CsA, unlike FK506 and rapamycin, does not bind to the intracellular immunophilin FK-binding protein (FKBP), the antagonism between these drugs is not simply due to competition for intracellular FKBP. Accordingly, RNA and protein stability analyses suggest inhibition by rapamycin at the translational level. Given the important role of LAT in initiating T cell activation, our data suggests that the effects of rapamycin, CsA and FK506 on T cell activation involve regulating early T cell signaling. These findings refine our understanding of the manifold effects of these immunosuppressants, thus providing insight into the drastic physiological contrasts observed between these drugs. PMID- 14565936 TI - Chimeric beta2 microglobulin/CD3zeta polypeptides expressed in T cells convert MHC class I peptide ligands into T cell activation receptors: a potential tool for specific targeting of pathogenic CD8(+) T cells. AB - CD8(+) T cells are key mediators of transplant rejection and graft-versus-host disease and contribute to the pathogenesis of autoimmune diseases. We tested whether TCR ligands can be converted into T cell activation receptors, redirecting genetically modified T cells at pathogenic CD8(+) T cells. For this purpose we exploited the ability of the non-polymorphic beta(2) microglobulin light chain to pair with all MHC class I heavy chains. In this report we describe the design and expression in a T cell hybridoma of two modalities of beta(2) microglobulin polypeptides, fused with the transmembrane and intracellular portion of CD3zeta chain. In the absence of a particular antigenic peptide, the chimeric product associates with different endogenous MHC class I heavy chains and triggers T cell activation upon heavy chain cross-linking. When an antigenic peptide is covalently attached to the N-terminus of the chimeric polypeptide, transfectants express high level of surface peptide-class I complexes and respond to antibodies and target T cells in a peptide-specific manner. Our results provide the basis for a universal genetic approach aimed at antigen-specific immunotargeting of pathogenic CD8(+) T cells. PMID- 14565938 TI - Root : shoot ratios, optimization and nitrogen productivity. AB - Plants respond to nitrogen availability by changing their root : shoot ratios. One hypothesis used to explain this allocation is that plants optimize their behaviour by maximizing their relative growth rate. The consequences of this hypothesis were investigated by formulating two models for root : shoot allocation, with and without explicit inclusion of maintenance respiration. The models also took into account that relative growth rate is a linear function of plant nitrogen concentration. The model without respiration gave qualitatively reasonable results when predictions were compared with observed results from growth experiments with birch and tomato. The explicit inclusion of maintenance respiration improved considerably the agreement between prediction and observation, and for birch was within the experimental accuracy. Further improvements will require additional details in the description of respiratory processes and the nitrogen uptake function. Plants growing under extreme nutrient stress may also optimize their behaviour with respect to other variables in addition to relative growth rate. PMID- 14565937 TI - CD75s is a marker of murine CD8(+) suppressor T cells. AB - We have previously described a monoclonal antibody, 984, which specifically recognizes murine CD8(+) suppressor T cells (Ts) but not CD8(+) cytolytic T lymphocytes (CTLs). Removal of 984(+) cells abrogates the suppressive effect of CD8(+) Ts generated either in vivo or in vitro while having no effect upon CTL. In this report, the molecules recognized by 984 are identified as 2-6 sialylated neolacto series gangliosides, which are members of the newly defined CD75s cluster. We proceed to demonstrate that like 984, a separate anti-CD75s antibody (CRIS-4), recognizes primary CD8(+) Ts cells. In addition, the 2,6 sialyltransferase responsible for the synthesis of the 984 epitope is identified, allowing the manipulation and study of the regulation of this epitope. This is the first report of CD75s on murine cells and the first report that delineates lymphocyte function based upon CD75s expression. In addition to contributing to the growing body of evidence that lineage dependent gangliosides are expressed by T lymphocytes, these findings suggest that CD8(+) CD75s(+) T lymphocytes represent a functionally distinct subset of CD8(+) T cells with negative regulatory function. PMID- 14565939 TI - Effects of external iron concentration upon seedling growth and uptake of Fe and phosphate by the common reed, Phragmites australis (Cav.) Trin ex. Steudel. AB - The objectives of this study were to determine whether, and to what degree, the aqueous iron concentration in the growing medium affects the growth of, and Fe uptake by, Phragmites australis, and whether the presence of iron in the growing environment affects the uptake of the essential element phosphate. The wetland macrophyte P. australis was grown under laboratory conditions in nutrient solution (0.31 mg L(-1) phosphate) containing a range of iron concentrations (0 50 mg L(-1) Fe). A threshold of iron concentration (1 mg L(-1)) was found, above which growth of P. australis was significantly inhibited. No direct causal relationship between iron content in aerial tissues and growth inhibition was found, which strongly suggests that iron toxicity cannot explain these results. Phosphate concentrations in aerial tissues were consistently sufficient for growth and development (2-3 % d. wt) despite significant variation in concentration of phosphate associated with roots. External Fe concentration had a significant effect on the growth of P. australis and on both Fe and phosphate concentrations associated with roots. However, neither direct toxicity nor phosphate deficiency could explain the reduction in growth above 1 mg L(-1) external Fe concentration PMID- 14565940 TI - Deficiency in neutrophil elastase does not impair neutrophil recruitment to inflamed sites. AB - To reach the sites of inflammation, neutrophils traverse the endothelium, its underlying basement membrane, and other barriers depending on the localization of the insulting agent. Whether neutrophil elastase (NE) plays a role in neutrophil recruitment to inflamed sites is still debatable. By exploiting mice deficient in NE (NE(-/-)), we sought to address this dilemma. We recruited neutrophils to the lungs or the peritoneum of wild-type (WT) or NE(-/-) mice by intranasal or intraperitoneal challenge with Pseudomonas aeruginosa or its lipopolysaccharide. At designated times post-inoculation (0, 4, 24, and 48 h), groups of mice were killed to assess changes in leukocyte counts and inflammatory responses. NE(-/-) and WT mice had normal circulating leukocyte numbers including neutrophils and changes in the hemograms in the setting of acute inflammation were indistinguishable. Analyses of lung tissues or fluids from the lungs and peritoneum found that regardless of the inflammatory model, the leukocyte counts including neutrophils and the inflammatory response were similar in NE(-/-) and WT mice at all time points. In vitro, neutrophils isolated from the lungs or the peritoneum of NE(-/-) and WT mice had comparable chemotactic and respiratory burst functions and migrated normally through Matrigel in response to various stimuli. Interestingly, preincubation of human peripheral blood neutrophils with NE physiologic inhibitors did not alter the migration of the cells through Matrigel. In sum, our findings present the first in vivo description that the absence of NE does not impair neutrophil recruitment to inflamed sites and that NE is not required for basement membrane transmigration of neutrophils. PMID- 14565941 TI - Human lung fibroblasts express interleukin-6 in response to signaling after mast cell contact. AB - Asthma is a chronic inflammatory disease of the airways. Mast cell-derived cytokines may mediate both airway inflammation and remodeling. It has also been shown that fibroblasts can be the source of proinflammatory cytokines. In the human airways, mast cell-fibroblast interactions may have pivotal effects on modulating inflammation. To study this further, we cocultured normal human lung fibroblasts (NHLF) with a human mast cell line (HMC-1) and assayed for production of interleukin (IL)-6, an important proinflammatory cytokine. When cultured together, NHLF/HMC-1 contact induced IL-6 secretion. Separation of HMC-1 and NHLF cells by a porous membrane inhibited this induction. HMC-1-derived cellular membranes caused an increase in IL-6 production in NHLF. Activation of p38 MAPK was also seen in cocultures by Western blot, whereas IL-6 production in cocultures was significantly inhibited by the p38 inhibitor SB203580. IL-6 production in cocultures was minimally inhibited by a chemical inhibitor of nuclear factor-kappaB (Bay11), indicating that nuclear factor-kappaB may have a minimal role in signaling IL-6 production in mast cell/fibroblasts cocultures. Blockade of inter-cellular adhesion molecule-1, tumor necrosis factor-RI, and surface IL-1beta with neutralizing antibodies failed to significantly decrease IL 6 production in our coculture, indicating that other receptor-ligand associations may be responsible for this activation. These novel studies reveal the importance of cell-cell interactions in the complex milieu of airway inflammation. PMID- 14565942 TI - Depletion of pulmonary intravascular macrophages inhibits acute lung inflammation. AB - Pulmonary intravascular macrophages (PIMs) are present in ruminants and horses. These species are highly sensitive to acute lung inflammation compared with non PIM-containing species such as rats and humans. There is evidence that rats and humans may also recruit PIMs under certain conditions. We investigated precise contributions of PIMs to acute lung inflammation in a calf model. First, PIMs were recognized with a combination of in vivo phagocytic tracer Monastral blue and postembedding immunohistology with anti-CD68 monoclonal antibody. Second, gadolinium chloride depleted PIMs within 48 h of treatment (P < 0.05). Finally, PIMs contain TNF-alpha, and their depletion reduces cells positive for IL-8 (P < 0.05) and TNF-alpha (P < 0.05) and histopathological signs of acute lung inflammation in calves infected with Mannheimia hemolytica. The majority of IL-8 positive inflammatory cells in lung septa of infected calves were platelets. Platelets from normal cattle contained preformed IL-8 that was released upon in vitro exposure to thrombin (P < 0.05). These novel data show that PIMs, as the source of TNF-alpha, promote recruitment of inflammatory cells including IL-8 containing platelets to stimulate acute inflammation and pathology in lungs. These data may also be relevant to humans due to our ability to recruit PIMs. PMID- 14565943 TI - DEP-induced fra-1 expression correlates with a distinct activation of AP-1 dependent gene transcription in the lung. AB - Recent studies indicate a potential role for Fra-1, a heterodimeric partner of activator protein (AP)-1, in toxicant-induced epithelial injury, repair, and cellular transformation. Here we have investigated the effects of diesel exhaust particles (DEP) on fra-1 expression in C10 cells, a murine lung epithelial cell line. DEP markedly upregulated fra-1, but not fra-2, expression. The increase in fra-1 mRNA expression correlated well with its protein- and DNA-binding activity. DNA-binding assays also revealed a predominant presence of Jun-B and Jun-D in the AP-1 complex. Interestingly, DEP did not alter Jun-B and Jun-D protein levels. Transcriptional analysis revealed that fra-1 induction is regulated in part at the transcriptional level. The -379 to +32 bp 5'-flanking region mediated this induction. Furthermore, inhibitors of ERK1/2, JNK1, and p38 mitogen-activated protein kinases (MAPKs) significantly suppressed DEP-stimulated fra-1 transcription, suggesting their involvement in the induction process. Consistent with this finding, DEP stimulated phosphorylation of ERK1/2, JNK1, and p38 MAPKs with a distinct activation pattern. Overexpression of Fra-1 downregulated c-Jun and Nrf2 enhanced AP-1- and ARE-mediated reporter gene expression, respectively. In contrast, Fra-1 had the opposite effect on matrix metalloproteinase (MMP)-9 promoter activity. In particular, it bound to the functional AP-1 site of the MMP 9 promoter after DEP stimulation. Consistent with this result, DEP also markedly upregulated MMP-9 promoter activity. Collectively, these findings suggest that fra-1 induction by DEP may play a role in selectively regulating gene expression involved in alveolar epithelial cell injury and repair. PMID- 14565944 TI - G protein-coupled receptor kinase 5 regulates airway responses induced by muscarinic receptor activation. AB - G protein-coupled receptors (GPCRs) transduce extracellular signals into intracellular events. The waning responsiveness of GPCRs in the face of persistent agonist stimulation, or desensitization, is a necessary event that ensures physiological homeostasis. GPCR kinases (GRKs) are important regulators of GPCR desensitization. GRK5, one member of the GRK family, desensitizes central M(2) muscarinic receptors in mice. We questioned whether GRK5 might also be an important regulator of peripheral muscarinic receptor responsiveness in the cardiopulmonary system. Specifically, we wanted to determine the role of GRK5 in regulating muscarinic receptor-mediated control of airway smooth muscle tone or regulation of cholinergic-induced bradycardia. Tracheal pressure, heart rate, and tracheal smooth muscle tension were measured in mice having a targeted deletion of the GRK5 gene (GRK5(-/-)) and littermate wild-type (WT) control mice. Both in vivo and in vitro results showed that the airway contractile response to a muscarinic receptor agonist was not different between GRK5(-/-) and WT mice. However, the relaxation component of bilateral vagal stimulation and the airway smooth muscle relaxation resulting from beta(2)-adrenergic receptor activation were diminished in GRK5(-/-) mice. These data suggest that M(2) muscarinic receptor-mediated opposition of airway smooth muscle relaxation is regulated by GRK5 and is, therefore, excessive in GRK5(-/-) mice. In addition, this study shows that GRK5 regulates pulmonary responses in a tissue- and receptor-specific manner but does not regulate peripheral cardiac muscarinic receptors. GRK5 regulation of airway responses may have implications in obstructive airway diseases such as asthma or chronic obstructive pulmonary disease. PMID- 14565945 TI - Pulmonary surfactant secretion in briefly cultured mouse type II cells. AB - There is little information on the regulation of surfactant secretion in mouse type II cells. We isolated type II cells from C57BL/6 and FVB mice, cultured them overnight, and then examined their response to known surfactant secretagogues. Secretion of phosphatidylcholine, surfactant protein (SP)-B and SP-C was stimulated by terbutaline, 5'-N-ethylcarboxyamidoadenosine (NECA), ATP, UTP, TPA, and ionomycin. Phosphatidylcholine secretion was increased approximately twofold by all agonists in both strains of mice. The response to terbutaline and NECA is the same as in rat type II cells, whereas the response to ATP, UTP, TPA, and ionomycin is considerably less. Secretion of SP-B and SP-C was increased sevenfold by terbutaline and threefold by ATP, effects similar to those in rat type II cells. The response to terbutaline was significantly decreased in type II cells from beta(2)-adrenergic receptor null mice. These data establish that briefly cultured type II cells provide a suitable model for investigation of surfactant secretion in normal and genetically altered mice. PMID- 14565946 TI - November--looking forward. PMID- 14565947 TI - Ground-based measurements of leaf area index: a review of methods, instruments and current controversies. AB - Leaf area index (LAI) is the total one-sided area of leaf tissue per unit ground surface area. It is a key parameter in ecophysiology, especially for scaling up the gas exchange from leaf to canopy level. It characterizes the canopy atmosphere interface, where most of the energy fluxes exchange. It is also one of the most difficult to quantify properly, owing to large spatial and temporal variability. Many methods have been developed to quantify LAI from the ground and some of them are also suitable for describing other structural parameters of the canopy. This paper reviews the direct and indirect methods, the required instruments, their advantages, disadvantages and accuracy of the results. Analysis of the literature shows that most cross-validations between direct and indirect methods have pointed to a significant underestimation of LAI with the latter techniques, especially in forest stands. The two main causes for the discrepancy, clumping and contribution of stem and branches, are discussed and some recent theoretical or technical solutions are presented as potential improvements to reduce bias or discrepancies. The accuracy, sampling strategy and spatial validity of the LAI measurements have to be assessed for quality assurance of both the measurement and the modelling purposes of all the LAI dependent ecophysiological and biophysical processes of canopies. PMID- 14565948 TI - Chlorophyll thermoluminescence of leaf discs: simple instruments and progress in signal interpretation open the way to new ecophysiological indicators. AB - Luminescence from photosynthetic material observed in darkness following illumination is a delayed fluorescence produced by a recombination of charge pairs stored in photosystem II, i.e. the back-reaction of photosynthetic charge separation. Thermoluminescence (TL) is a technique consisting of a rapid cooling followed by the progressive warming of a preilluminated sample to reveal the different types of charge pairs as successive emission bands, which are resolved better than the corresponding decay phases recorded at constant temperatures. Progress in thermoelectric Peltier elements and in compact light detectors made the development of simple, affordable and transportable instruments possible. These instruments take advantage of multifurcated light guides for combined TL, fluorescence and absorbance/reflectance measurements. Meanwhile, experiments on unfrozen leaf discs, with excitation by single turn-over flashes or far red light and infiltration by specific inhibitors/uncouplers, have led to a better understanding of in vivo TL signals. Much like chlorophyll fluorescence and in a complementary way, TL in the 0-60 degrees C temperature range not only informs on the state of photosystem II in leaf tissues and its possible alterations, but also gives a broader insight into the energetic state inside the chloroplast by probing (1) the light-induced or dark-stable thylakoid proton gradient through the protonation of the Mn oxygen-evolving complex, (2) the induction of cyclic/chlororespiratory electron flow towards the plastoquinone pool, (3) the [NADPH+ATP] assimilatory potential. By a different mechanism, warming above 60 degrees C without preillumination reveals chemiluminescence high temperature thermoluminescence (HTL) bands due to the radiative thermolysis of peroxides, which are indicators of oxidative stress in leaves. PMID- 14565949 TI - The role of ethylene during the infection of Nicotiana tabacum by Colletotrichum destructivum. AB - Two periods of increased ethylene production were observed after inoculation of Nicotiana tabacum by Colletotrichum destructivum. This pathogen exhibits an intracellular hemibiotrophic infection process, with a biotrophic phase followed by a necrotrophic phase. Ethylene production first increased during the biotrophic phase with a peak at 24 h before the necrotrophic phase. A second increase in ethylene occurred late in the necrotrophic phase when the lesions were expanding. Two different 1-aminocyclopropane-1-carboxylic acid synthase genes showed increased expression after the first ethylene peak with a maximum at 24 h before the second ethylene increase. Expression of an 1-aminocyclopropane-1 carboxylic acid oxidase (ACO) gene increased during the first ethylene peak and then declined at the beginning of the second ethylene increase. A second ACO gene showed relatively little change in expression during infection with slightly higher expression at 24 h before the second ethylene increase, and a third ACO gene showed a progressive decline in expression with a major decrease occurring before the second ethylene increase. Inoculation of ethylene-insensitive tobacco with C. destructivum revealed that it was more susceptible than the wild type. The changes in ethylene production and associated gene expression as well as the increased disease susceptibility of ethylene-insensitive tobacco indicate that ethylene plays a role in this interaction, perhaps as a signalling molecule to trigger defense mechanisms. PMID- 14565950 TI - Effect of P-deficiency on photoassimilate partitioning and rhythmic changes in fruit and stem diameter of tomato (Lycopersicon esculentum) during fruit growth. AB - Tomato (Lycopersicon esculentum) plants were grown in liquid culture inside the greenhouse of Hiroshima University, Japan. At the first fruiting stage, P was withdrawn from the rooting medium for a period of 19 d and its effect was studied on photosynthesis, stomatal conductance, transpiration, partitioning of 13C and 15N, P contents of various organs, and changes in stem and fruit diameter of the plant in order to identify the mechanism of resource management on the part of the plant at low P. Compared to the control, P-deficiency treatment decreased biomass growth of all organs except the roots. The treatment also depressed leaf photosynthesis, stomatal conductance and diameter of fruit and stem after a lag period of about 1 week. The stem diameter of the plant shrank during daytime and expanded during the night; the adverse effect of P-deficiency on stem diameter change was more evident during the night than the day. The circadian rhythm in fluctuations of diameter was less manifested in the fruit compared with the stem. P-deficiency induced daytime shrinkage and reduced night expansion of fruit. However, within the plant, P-deficiency encouraged partitioning of 13C, 15N and P into the fruit at the cost of autotrophic organs such as leaves and the upper parts of the stem. The results were discussed in the light of a plausible effect of P-deficiency on water relations of the plant. It is concluded that, in spite of the preference in partitioning of C and N received within the plant parts, assimilate flow into the fruit is limited at low-P compared with the control, owing to the restriction in fruit expansion. PMID- 14565951 TI - Screening of inbred lines to develop a thermotolerant sunflower hybrid using the temperature induction response (TIR) technique: a novel approach by exploiting residual variability. AB - Plants, when exposed to sub-lethal stress (induction stress), develop the ability to withstand severe temperatures and this phenomenon is often referred to as acquired thermotolerance. Earlier it was reported that induction stress alters gene expression and brings greater adaptation to heat stress and that the genetic variability in thermotolerance is only seen upon induction stress. Based on this concept, the temperature induction response (TIR) technique has been developed to identify thermotolerant lines. By following the TIR technique, sunflower hybrid KBSH-1 parents were screened for high temperature tolerance. Seedlings of parental lines including CMS 234 A, CMS 234 B and 6 D-1 showed considerable genetic variability for thermotolerance and it was attributed to the expression of existing residual variability for stress responses. Thus, the existing variability forms the basis for identifying thermotolerant lines. The identified parental inbred lines were selected and established in the field and crossed to get F1 hybrid seeds. The KBSH-1 hybrid developed from selected variants of parental lines was compared with the original KBSH-1 for thermotolerance. The selected KBSH-1 was more tolerant compared with the original hybrid both at the seedling as well as at the plant level. The physiological and molecular basis of thermotolerance was studied in the KBSH-1 original and the hybrid developed from selected variants of parental lines. The selected hybrid exhibited high tolerance to Menadione (naphthoquinone)-induced oxidative stress. Even the methyl viologen induced oxidative stress damage was relatively less in the selected hybrid population. The selected hybrid also showed enhanced expression of the heat shock proteins HSP 90 and HSP 104 and also accumulated higher levels of the heat shock transcription factor HSFA. PMID- 14565952 TI - Megakaryoblastic leukemia-1/2, a transcriptional co-activator of serum response factor, is required for skeletal myogenic differentiation. AB - Serum response factor (SRF) is required for the expression of a wide variety of muscle-specific genes that are expressed upon differentiation and is thus required for both striated and smooth muscle differentiation in addition to its role in regulating growth factor-inducible genes. A heart and smooth muscle specific SRF co-activator, myocardin, has been shown to be required for cardiac development and smooth muscle differentiation. However, no such co-factors of SRF have been identified in the skeletal myogenic differentiation program. Myocardin and the related transcription factor megakaryoblastic leukemia-1 (MKL1/MAL/MRTF A) can strongly potentiate the activity of SRF. Here we report the cloning of the third member of the myocardin/MKL family in humans, MKL2. MKL2 binds to and activates SRF similar to myocardin and MKL1. To determine the role of these factors in skeletal myogenic differentiation we used a dominant negative MKL2 to show that the MKL family of proteins is required for skeletal myogenic differentiation. Expression of the dominant negative protein in C2C12 skeletal myoblasts blocked the differentiation-induced expression of the SRF target genes skeletal alpha-actin and alpha-myosin heavy chain and blocked differentiation of the myoblasts to myotubes in vitro. C2C12 cells express both MKL1 and MKL2, but not myocardin, implicating MKL1 and/or MKL2 in the requirement for skeletal myogenic differentiation. MKL1 was predominantly cytoplasmic in C2C12 cells, with a small amount in the nucleus, however, no movement of MKL1 to the nucleus was observed upon differentiation. PMID- 14565954 TI - Janus kinase 2 and calcium are required for angiotensin II-dependent activation of steroidogenic acute regulatory protein transcription in H295R human adrenocortical cells. AB - Angiotensin II- and K+-stimulated aldosterone production in the adrenocortical glomerulosa cells requires induction of the steroidogenic acute regulatory protein (StAR). While both agents activate Ca2+ signaling, the mechanisms leading to aldosterone synthesis are distinct, and the angiotensin II response cannot be mimicked by K+. We previously reported that StAR mRNA levels and promoter reporter gene activity in transiently transfected H295R human adrenocortical cells were stimulated by angiotensin II but not by K+ treatment. The current study focused on identifying signaling pathways activated by angiotensin II that contribute to StAR transcriptional activation. We show that the angiotensin II stimulated transcriptional activation of StAR was dependent upon influx of external calcium and requires protein kinase C activation. Furthermore we describe for the first time that the Janus tyrosine kinase family member, JAK2, was activated by angiotensin II treatment of H295R cells. Treatment of the cells with AG490, a selective inhibitor of JAK2, blocked JAK2 activation and StAR reporter gene activity and inhibited steroid production. Taken together these studies describe a novel pathway controlling StAR expression and steroidogenesis in adrenocortical cells. PMID- 14565953 TI - Molecular cloning, genomic structure, and expression analysis of MUC20, a novel mucin protein, up-regulated in injured kidney. AB - Immunoglobulin A nephropathy (IgAN) is the most common primary glomerulonephritis in the world. Here, we identify a cDNA encoding a novel mucin protein, shown previously to be up-regulated in IgAN patients, from a human kidney cDNA library. This protein contains a mucin tandem repeat of 19 amino acids consisting of many threonine, serine, and proline residues and likely to be extensively O glycosylated; thus, this gene was classified in the mucin family and named MUC20. The human MUC20 gene contains at least four exons and is localized close to MUC4 on chromosome 3q29. We found variations in repeat numbers in the mucin tandem domain, suggesting polymorphism of this region. Northern blot and reverse transcription-PCR analyses revealed that human MUC20 mRNA was expressed most highly in kidney and moderately in placenta, colon, lung, prostate, and liver. Immunohistochemical analysis of human kidney revealed that MUC20 protein was localized in the proximal tubules. Immunoblotting analysis of MUC20 proteins produced in Madin-Darby canine kidney and HEK293 cells indicated the localization of MUC20 protein in a membrane fraction and extensive posttranslational modification. Immunoelectron microscopy of MUC20-producing Madin-Darby canine kidney cells demonstrated that MUC20 protein was localized on the plasma membrane. Expression of MUC20 mRNA in a human kidney cell line was up-regulated by tumor necrosis factor-alpha, phorbol 12-myristate 13-acetate, or lipopolysaccharide. Two species of MUC20 mRNA (hMUC20-L and hMUC20-S), resulting from alternative transcription, were identified in human tissue, whereas only one variant was observed in mouse tissues. Mouse MUC20 mRNA was expressed in the epithelial cells of proximal tubules, and the expression increased dramatically with the progression of lupus nephritis in the kidney of MRL/MpJ-lpr/lpr mice. Moreover, the expression of mouse MUC20 was augmented in renal tissues acutely injured by cisplatin or unilateral ureteral obstruction. These characteristics suggest that the production of MUC20 is correlated with development and progression of IgAN and other renal injuries. PMID- 14565955 TI - A novel AP-2 adaptor interaction motif initially identified in the long-splice isoform of synaptojanin 1, SJ170. AB - Phosphoinositides play a fundamental role in clathrin-coat assembly at the cell surface. Several endocytic components and accessory factors contain independently folded phosphoinositide-binding modules that facilitate, in part, membrane placement at the bud site. As the clathrin-coat assembly process progresses toward deeply invaginated buds, focally synthesized phosphoinositides are dephosphorylated, principally through the action of the phosphoinositide polyphosphatase synaptojanin 1. Failure to catabolize polyphosphoinositides retards the fission process and endocytic activity. The long-splice isoform of synaptojanin 1, termed SJ170, contains a carboxyl-terminal extension that harbors interaction motifs for engaging several components of the endocytic machinery. Here, we demonstrate that in addition to DPF and FXDXF sequences, the SJ170 carboxyl terminus contains a novel AP-2 binding sequence, the WXXF motif. The WXXF sequence engages the independently folded alpha-subunit appendage that projects off the heterotetrameric AP-2 adaptor core. The endocytic protein kinases AAK1 and GAK also contain functional WXX(FW) motifs in addition to two DPF repeats, whereas stonin 2 harbors three tandem WXXF repeats. Each of the discrete SJ170 adaptor-interaction motifs bind to appendages relatively weakly but, as tandemly arrayed within the SJ170 extension, can cooperate to bind bivalent AP-2 with good apparent affinity. These interactions likely contribute to the appropriate targeting of certain endocytic components to clathrin bud sites assembling at the cell surface. PMID- 14565956 TI - Critical role of the transcriptional repressor neuron-restrictive silencer factor in the specific control of connexin36 in insulin-producing cell lines. AB - Connexin36 (Cx36) is specifically expressed in neurons and in pancreatic beta cells. Cx36 functions as a critical regulator of insulin secretion and content in beta-cells. In order to identify the molecular mechanisms that control the beta cell expression of Cx36, we initiated the characterization of the human 5' regulatory region of the CX36 gene. A 2043-bp fragment of the human CX36 promoter was identified from a human BAC library and fused to a luciferase reporter gene. This promoter region was sufficient to confer specific expression to the reporter gene in insulin-secreting cell lines. Within this 5' regulatory region, a putative neuron-restrictive silencer element conserved between rodent and human species was recognized and binds the neuron-restrictive silencing factor (NRSF/REST). This factor is not expressed in insulin-secreting cells and neurons; it functions as a potent repressor through the recruitment of histone deacetylase to the promoter of neuronal genes. The NRSF-mediated repression of Cx36 in HeLa cells was abolished by trichostatin A, confirming the functional importance of histone deacetylase activity. Ectopic expression, by viral gene transfer, of NRSF/REST in different insulin-secreting beta-cell lines induced a marked reduction in Cx36 mRNA and protein content. Moreover, mutations in the Cx36 neuron-restrictive silencer element relieved the low transcriptional activity of the human CX36 promoter observed in HeLa cells and in INS-1 cells expressing NRSF/REST. The data showed that cx36 gene expression in insulin-producing beta cell lines is strictly controlled by the transcriptional repressor NRSF/REST indicating that Cx36 participates to the neuronal phenotype of the pancreatic beta-cells. PMID- 14565957 TI - Antagonism of rat beta-cell voltage-dependent K+ currents by exendin 4 requires dual activation of the cAMP/protein kinase A and phosphatidylinositol 3-kinase signaling pathways. AB - Antagonism of voltage-dependent K+ (Kv) currents in pancreatic beta-cells may contribute to the ability of glucagon-like peptide-1 (GLP-1) to stimulate insulin secretion. The mechanism and signaling pathway regulating these currents in rat beta-cells were investigated using the GLP-1 receptor agonist exendin 4. Inhibition of Kv currents resulted from a 20-mV leftward shift in the voltage dependence of steady-state inactivation. Blocking cAMP or protein kinase A (PKA) signaling (Rp-cAMP and H-89, respectively) prevented the inhibition of currents by exendin 4. However, direct activation of this pathway alone by intracellular dialysis of cAMP or the PKA catalytic subunit (cPKA) could not inhibit currents, implicating a role for alternative signaling pathways. A number of phosphorylation sites associated with phosphatidylinositol 3 (PI3)-kinase activation were up-regulated in GLP-1-treated MIN6 insulinoma cells, and the PI3 kinase inhibitor wortmannin could prevent antagonism of beta-cell currents by exendin 4. Antagonists of Src family kinases (PP1) and the epidermal growth factor (EGF) receptor (AG1478) also prevented current inhibition by exendin 4, demonstrating a role for Src kinase-mediated trans-activation of the EGF tyrosine kinase receptor. Accordingly, the EGF receptor agonist betacellulin could replicate the effects of exendin 4 in the presence of elevated intracellular cAMP. Downstream, the PKCzeta pseudosubstrate inhibitor could prevent current inhibition by exendin 4. Therefore, antagonism of beta-cell Kv currents by GLP-1 receptor activation requires both cAMP/PKA and PI3 kinase/PKCzeta signaling via trans-activation of the EGF receptor. This represents a novel dual pathway for the control of Kv currents by G protein-coupled receptors. PMID- 14565958 TI - Expression and function of gonadotropin-releasing hormone (GnRH) receptor in human olfactory GnRH-secreting neurons: an autocrine GnRH loop underlies neuronal migration. AB - Olfactory neurons and gonadotropin-releasing hormone (GnRH) neurons share a common origin during organogenesis. Kallmann's syndrome, clinically characterized by anosmia and hypogonadotropic hypogonadism, is due to an abnormality in the migration of olfactory and GnRH neurons. We recently characterized the human FNC B4 cell line, which retains properties present in vivo in both olfactory and GnRH neurons. In this study, we found that FNC-B4 neurons expressed GnRH receptor and responded to GnRH with time- and dose-dependent increases in GnRH gene expression and protein release (up to 5-fold). In addition, GnRH and its analogs stimulated cAMP production and calcium mobilization, although at different biological thresholds (nanomolar for cAMP and micromolar concentrations for calcium). We also observed that GnRH triggered axon growth, actin cytoskeleton remodeling, and a dose-dependent increase in migration (up to 3-4-fold), whereas it down regulated nestin expression. All these effects were blocked by a specific GnRH receptor antagonist, cetrorelix. We suggest that GnRH, secreted by olfactory neuroblasts, acts in an autocrine pattern to promote differentiation and migration of those cells that diverge from the olfactory sensory lineage and are committed to becoming GnRH neurons. PMID- 14565959 TI - Respiratory syncytial virus up-regulates TLR4 and sensitizes airway epithelial cells to endotoxin. AB - Airway epithelial cells are unresponsive to endotoxin (lipopolysaccharide (LPS)) exposure under normal conditions. This study demonstrates that respiratory syncytial virus (RSV) infection results in increased sensitivity to this environmental exposure. Infection with RSV results in increased expression of Toll-like receptor (TLR) 4 mRNA, protein, and increased TLR4 membrane localization. This permits significantly enhanced LPS binding to the epithelial monolayer that is blocked by disruption of the Golgi. The increased TLR4 results in an LPS-induced inflammatory response as demonstrated by increased mitogen activated protein (MAP) kinase activity, IL-8 production, and tumor necrosis factor alpha production. RSV infection also allowed for tumor necrosis factor alpha production subsequent to TLR4 cross-linking with an immobilized antibody. These data suggest that RSV infection sensitizes airway epithelium to a subsequent environmental exposure (LPS) by altered expression and membrane localization of TLR4. The increased interaction between airway epithelial cells and LPS has the potential to profoundly alter airway inflammation. PMID- 14565960 TI - SH2-B is a positive regulator of nerve growth factor-mediated activation of the Akt/Forkhead pathway in PC12 cells. AB - To gain insight into the mechanism by which the adapter protein SH2-B promotes nerve growth factor (NGF)-mediated neuronal differentiation and survival, the effect of SH2-B on the serine/threonine kinase Akt/protein kinase B and downstream effector proteins was examined. PC12 cells stably overexpressing SH2 Bbeta, which exhibit enhanced NGF-induced neuronal differentiation compared with control cells, showed enhanced and prolonged NGF-induced phosphorylation of Akt on Ser473 and Akt enzymatic activity. Surprisingly, NGF-induced phosphorylation of Akt on Ser473 and Akt activity were not altered in cells overexpressing SH2 Bbeta(R555E) with a defective SH2 domain, despite the ability of the overexpressed SH2-Bbeta(R555E) to block NGF-induced differentiation. Consistent with SH2-Bbeta enhancing the activity of Akt, cells overexpressing SH2-Bbeta but not SH2-Bbeta(R555E) exhibited increased and/or prolonged phosphorylation of the pro-apoptotic Akt effector proteins, glycogen synthase kinase-3, and forkhead transcription factors, FKHRL1/FOXO3 and FKHR/FOXO1. Immunolocalization studies indicated that, although ectopically expressed FKHR was primarily concentrated in the cytoplasm of control cells and cells transiently overexpressing SH2-Bbeta, it was concentrated in the nucleus of cells transiently overexpressing SH2 Bbeta(R555E). Similarly, SH2-Bbeta stimulated the accumulation of FKHR in the cytoplasm of 293T and COS-7 cells, whereas SH2-Bbeta(R555E) enhanced its accumulation in the nucleus. In PC12 cells stably expressing forms of SH2-Bbeta, SH2-Bbeta mimicked the ability of NGF to promote redistribution of FKHR to the cytoplasm whereas SH2-Bbeta(R555E) blocked this effect of NGF. Taken together, these data indicate that SH2-B is a positive regulator of NGF-mediated activation of the Akt/Forkhead pathway. PMID- 14565961 TI - Human telomerase reverse transcriptase motifs required for elongation of a telomeric substrate. AB - The reverse transcriptase telomerase copies an internal RNA template to synthesize telomeric simple-sequence repeats. In the cellular context, telomerase must elongate its few intended substrates (authentic chromosome ends) without spurious activity on other potential substrates (chromosome ends created by damage, repair, or recombination). Many mechanisms have been proposed to account for the biological substrate specificity of telomerase, with most models focusing on protein-protein interactions between telomerase and telomeric chromatin. Telomerase activity assays testing the elongation of model oligonucleotide substrates have revealed that in addition to hybridization with the RNA template, optimal DNA substrates also engage telomerase protein-based interaction sites. The physiological significance of these non-template interaction sites has not been established. We used in vivo reconstitution to assemble telomerase enzymes with variant telomerase reverse transcriptase proteins. Several telomerase enzyme variants retained a wild-type level of catalytic function in vitro when assayed using an artificial sequence substrate but exhibited reduced activity on a more physiological telomeric-sequence substrate. Telomerases that demonstrated this defect in telomeric substrate usage in vitro also failed to support telomere length maintenance in vivo. Our findings suggest that non-template interactions of the telomerase ribonucleoprotein with telomeric DNA play a critical role in supporting telomerase function on its appropriate cellular substrates. PMID- 14565962 TI - Adaptations in metabolic capacity of rat soleus after paralysis. AB - To determine whether long-term reductions in neuromuscular activity result in alterations in metabolic capacity, the activities of oxidative, i.e., succinate dehydrogenase (SDH) and citrate synthase (CS), and glycolytic, i.e., alpha glycerophosphate dehydrogenase (GPD), enzyme markers were quantified in rat soleus muscles 1, 3, and 6 mo after a complete spinal cord transection (ST). In addition, the proportional content of lactate dehydrogenase (LDH) isozymes was used as a marker for oxidative and glycolytic capacities. The myosin heavy chain (MHC) isoform content of a fiber served as a marker of phenotype. In general, MHC isoforms shifted from MHC1 toward MHC2, particularly MHC2x, after ST. Mean SDH and CS activities were higher in ST than control at all time points. The elevated SDH and CS activities were indicative of an enhanced oxidative capacity. GPD activities were higher in ST than control rats at all time points. The increase in activity of SDH was larger than GPD. Thus the GPD-to-SDH (glycolytic-to oxidative) ratio was decreased after ST. Compared with controls, total LDH activity increased transiently, and the LDH isozyme profile shifted from LDH-1 toward LDH-5, indicative of an enhanced glycolytic capacity. Combined, these results indicate that 1) the metabolic capacities of soleus fibers were not compromised, but the interrelationships among oxidative and glycolytic capacity and MHC content were apparently dissociated after ST; 2) enhancements in oxidative and glycolytic enzyme activities are not mutually exclusive; and 3) chronic reductions in skeletal muscle activity do not necessarily result in a reduced oxidative capacity. PMID- 14565964 TI - Chronic O2 exposure in the newborn rat results in decreased pulmonary arterial nitric oxide release and altered smooth muscle response to isoprostane. AB - Chronic oxygen exposure in the newborn rat results in lung isoprostane formation, which may contribute to the pulmonary hypertension evident in this animal model. The purpose of this study was to investigate the pulmonary arterial smooth muscle responses to 8-iso-prostaglandin F(2alpha) (8-iso-PGF(2a)) in newborn rats exposed to 60% O2 for 14 days. Because, in the adult rat, 8-iso-PGF(2alpha) may have a relaxant effect, mediated by nitric oxide (NO), we also sought to evaluate the pulmonary arterial NO synthase (NOS) protein content and NO release in the newborn exposed to chronic hyperoxia. Compared with air-exposed control animals, 8-iso-PGF(2a) induced a significantly greater force (P < 0.01) and reduced (P < 0.01) relaxation of precontracted pulmonary arteries in the 60% O2-treated animals. These changes were reproduced in control pulmonary arteries by NOS blockade by using NG-nitro-L-arginine methyl ester. Pulmonary arterial endothelial NOS was unaltered, but the inducible NOS protein content was significantly decreased (P < 0.01) in the experimental group. Pulmonary (P < 0.05) and aortic (P < 0.01) tissue ex vivo NO accumulation was significantly reduced in the 60% O2-treated animals. We speculate that impaired pulmonary vascular tissue NO metabolism after chronic O2 exposure potentiates 8-iso PGF(2alpha)-induced vasoconstriction in the newborn rat, thus contributing to pulmonary hypertension. PMID- 14565963 TI - Keratinocyte growth factor transiently alters pulmonary function in rats. AB - Keratinocyte growth factor (KGF) is a mitogen for pulmonary epithelial cells. Intratracheal administration of KGF to adult rats results in alveolar epithelial type II and bronchiolar epithelial cell proliferation. While cellular responses to KGF have been intensively studied, functional consequences regarding lung function are unknown. Therefore, in this study, we sought to investigate whether KGF alters pulmonary function variables. Rats received either recombinant human KGF (rHuKGF) (5 mg/kg) or vehicle intratracheally. Before and on days 3 and 7 after treatment, pulmonary function was determined by body plethysmography. Subsequently, lung histological changes were quantified. rHuKGF induced a transient proliferation of alveolar and bronchiolar epithelial cells. The extent of type II cell hyperplasia was significantly correlated with a transient reduction in tidal volume and an increase in breathing frequency. In addition, quasi-static compliance, total lung capacity, and vital capacity were reduced after rHuKGF instillation, suggesting the development of a transitory restrictive lung disorder. Moreover, reduced expiratory flow rates and forced expiratory volumes, as well as increased functional residual capacity after rHuKGF but not vehicle, suggest obstructive lung function changes. In conclusion, the induction of alveolar and bronchiolar epithelial cell proliferation by KGF is paralleled by moderate functional consequences that should be taken into account when the therapeutic potential of KGF is tested. PMID- 14565965 TI - Functional magnetic resonance signal changes in neural structures to baroreceptor reflex activation. AB - The sequence of neural responses to exogenous arterial pressure manipulation remains unclear, especially for extramedullary sites. We used functional magnetic resonance imaging procedures to visualize neural responses during pressor (phenylephrine) and depressor (sodium nitroprusside) challenges in seven isoflurane-anesthetized adult cats. Depressor challenges produced signal intensity declines in multiple cardiovascular-related sites in the medulla, including the nucleus tractus solitarius, and caudal and rostral ventrolateral medulla. Signal decreases also emerged in the cerebellar vermis, inferior olive, dorsolateral pons, and right insula. Rostral sites, such as the amygdala and hypothalamus, increased signal intensity as arterial pressure declined. In contrast, arterial pressure elevation elicited smaller signal increases in medullary regions, the dorsolateral pons, and the right insula and signal declines in regions of the hypothalamus, with no change in deep cerebellar areas. Responses to both pressor and depressor challenges were typically lateralized. In a subset of animals, barodenervation resulted in rises and falls of blood pressure that were comparable to these resulting from the pharmacological challenges but different regional neural responses, indicating that the regional signal intensity responses did not derive from global perfusion effects but from baroreceptor mediation of central mechanisms. The findings demonstrate widespread lateralized distribution of neural sites responsive to blood pressure manipulation. The distribution and time course of neural responses follow patterns associated with early and late compensatory reactions. PMID- 14565966 TI - Strength training reduces force fluctuations during anisometric contractions of the quadriceps femoris muscles in old adults. AB - The greater fluctuations in motor output that are often exhibited by old adults can be reduced with strength training. The purpose of the study was to determine the effect of strength and steadiness training by old adults on fluctuations in force and position during voluntary contractions with the quadriceps femoris muscle. Healthy old adults (65-80 yr) completed 16 wk of heavy-load (80% of maximum, n = 11) strength training, heavy-load steadiness training (n = 6), or no training (n = 9). Steadiness training required subjects to match the angular displacement about the knee joint to a constant-velocity template. The Heavy-Load group experienced a 5.5% increase in muscle volume, a 25% increase in maximal voluntary contraction force, and a 26% increase in the one-repetition (1-RM) load. The Heavy-Load Steady group experienced increases of 11.5, 31, and 36%, respectively. The maximal electromyogram signal of quadriceps femoris increased by 51% in the two training groups. The coefficient of variation (CV) for force during submaximal isometric contractions did not change with training for any group. Although both training groups also experienced a reduction in CV for force during anisometric contractions with a 50% 1-RM load, the standard deviation of position did not change with time for any group. The Heavy-Load Steady group also experienced a reduction in CV for force during the training contractions performed with the 80% 1-RM load. Thus strength training reduced the force fluctuations of the quadriceps femoris muscles during anisometric contractions but not during isometric contractions. PMID- 14565967 TI - Alterations in the expression of mRNAs and proteins that code for species relevant to eIF2B activity after an acute bout of resistance exercise. AB - The focus of the study described herein was to examine the relative expression levels of mRNAs and proteins relevant to the regulation of translational initiation, and hence protein synthesis, in the time course after an acute bout of resistance exercise in male Sprague-Dawley rats. Significant increases in the relative abundance of the mRNAs coding for the epsilon (33%) and gamma (26%) subunits of eukaryotic initiation factor (eIF) 2B were observed 48 h after the exercise bout. Furthermore, the mRNA coding for the delta subunit of eIF2B was also significantly increased, both 24 h (46%) and 48 h (44%) postexercise. There was a relative decrease in three eIF2Bepsilon kinase mRNAs, namely sequences coding for glycogen synthase kinase 3beta (49%), casein kinase I (48%), and casein kinase II (42%) 48 h into the recovery period. Additionally, there was a significant decrease in expression of the mRNAs coding for eIF2alpha (28% 24 h postexercise) and one of its regulatory kinases, double-stranded RNA-activated protein kinase (33% 48 h postexercise). Finally, an increase in eIF2B total protein (124%) was observed within 3 h postexercise. These results suggest that there may be rapid translational regulation of mRNAs coding for species relevant to translational initiation after an acute bout of resistance exercise. Furthermore, transcription of these mRNAs is altered further into the recovery period, and this might play a role in protein synthetic capacity on subsequent bouts of resistance exercise. PMID- 14565968 TI - Transcriptional adaptations of lipid metabolism in tibialis anterior muscle of endurance-trained athletes. AB - It was hypothesized that transcriptional reprogramming is involved in the structural and functional adaptations of lipid metabolism in human tibialis anterior muscle (TA) from endurance-trained male subjects. RT-PCR experiments demonstrated a significant upregulation of the mRNA level of key enzymes involved in 1) lipolytic mobilization of fatty acids (FA) from intramyocellular lipid (IMCL) stores via hormone-sensitive lipase (LIPE), 2) intramyocellular FA transport via muscle fatty acid binding protein (FABP3), and 3) oxidative phosphorylation (cytochrome c oxidase I, COI), in TA of endurance-trained vs. untrained subjects. In contrast, mRNAs for factors involved in glycolysis (muscle 6-phosphofructokinase, PFKM), intramyocellular storage of FA (diacylglycerol O acyltransferase 1, DGAT), and beta-oxidation (long-chain acyl-coenzyme A dehydrogenase, ACADL) were invariant between TA of trained and untrained subjects. Correlation analysis identified an association of LIPE with FABP3 and LPL (lipoprotein lipase) mRNA levels and indicated coregulation of the transcript level for LIPE, FABP3, and COI with the level of mRNA encoding peroxisome proliferator-activated receptor-alpha (PPAR-alpha), the master regulator of lipid metabolism. Moreover, a significant correlation existed between LPL mRNA and the absolute rate of IMCL repletion determined by magnetic resonance spectroscopy after exhaustive exercise. Additionally, the LIPE mRNA level correlated with ultrastructurally determined IMCL content and mitochondrial volume density. The present data point to a training-induced, selective increase in mRNA levels of enzymes which are involved in metabolization of intramuscular FA, and these data confirm the well-established phenomenon of enhanced lipid utilization during exercise at moderate intensity in muscles of endurance-trained subjects. PMID- 14565970 TI - rsly1 binding to syntaxin 5 is required for endoplasmic reticulum-to-Golgi transport but does not promote SNARE motif accessibility. AB - Although some of the principles of N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) function are well understood, remarkably little detail is known about sec1/munc18 (SM) protein function and its relationship to SNAREs. Popular models of SM protein function hold that these proteins promote or maintain an open and/or monomeric pool of syntaxin molecules available for SNARE complex formation. To address the functional relationship of the mammalian endoplasmic reticulum/Golgi SM protein rsly1 and its SNARE binding partner syntaxin 5, we produced a conformation-specific monoclonal antibody that binds only the available, but not the cis-SNARE-complexed nor intramolecularly closed form of syntaxin 5. Immunostaining experiments demonstrated that syntaxin 5 SNARE motif availability is nonuniformly distributed and focally regulated. In vitro endoplasmic reticulum-to-Golgi transport assays revealed that rsly1 was acutely required for transport, and that binding to syntaxin 5 was absolutely required for its function. Finally, manipulation of rsly1-syntaxin 5 interactions in vivo revealed that they had remarkably little impact on the pool of available syntaxin 5 SNARE motif. Our results argue that although rsly1 does not seem to regulate the availability of syntaxin 5, its function is intimately associated with syntaxin binding, perhaps promoting a later step in SNARE complex formation or function. PMID- 14565969 TI - Disease-related myotubularins function in endocytic traffic in Caenorhabditis elegans. AB - MTM1, MTMR2, and SBF2 belong to a family of proteins called the myotubularins. X linked myotubular myopathy, a severe congenital disorder characterized by hypotonia and generalized muscle weakness in newborn males, is caused by mutations in MTM1 (Laporte et al., 1996). Charcot-Marie-Tooth types 4B1 and 4B2 are severe demyelinating neuropathies caused by mutations in MTMR2 (Bolino et al., 2000) and SBF2/MTMR13 (Senderek et al., 2003), respectively. Although several myotubularins are known to regulate phosphoinositide-phosphate levels in cells, little is known about the actual cellular process that is defective in patients with these diseases. Mutations in worm MTM-6 and MTM-9, myotubularins belonging to two subgroups, disorganize phosphoinositide 3-phosphate localization and block endocytosis in the coelomocytes of Caenorhabditis elegans. We demonstrate that MTM-6 and MTM-9 function as part of a complex to regulate an endocytic pathway that involves the Arf6 GTPase, and we define protein domains required for MTM-6 activity. PMID- 14565971 TI - Mhr1p-dependent concatemeric mitochondrial DNA formation for generating yeast mitochondrial homoplasmic cells. AB - Mitochondria carry many copies of mitochondrial DNA (mtDNA), but mt-alleles quickly segregate during mitotic growth through unknown mechanisms. Consequently, all mtDNA copies are often genetically homogeneous within each individual ("homoplasmic"). Our previous study suggested that tandem multimers ("concatemers") formed mainly by the Mhr1p (a yeast nuclear gene-encoded mtDNA recombination protein)-dependent pathway are required for mtDNA partitioning into buds with concomitant monomerization. The transmission of a few randomly selected clones (as concatemers) of mtDNA into buds is a possible mechanism to establish homoplasmy. The current study provides evidence for this hypothesis as follows: the overexpression of MHR1 accelerates mt-allele-segregation in growing heteroplasmic zygotes, and mhr1-1 (recombination-deficient) causes its delay. The mt-allele-segregation rate correlates with the abundance of concatemers, which depends on Mhr1p. In G1-arrested cells, concatemeric mtDNA was labeled by [14C]thymidine at a much higher density than monomers, indicating concatemers as the immediate products of mtDNA replication, most likely in a rolling circle mode. After releasing the G1 arrest in the absence of [14C]thymidine, the monomers as the major species in growing buds of dividing cells bear a similar density of 14C as the concatemers in the mother cells, indicating that the concatemers in mother cells are the precursors of the monomers in buds. PMID- 14565972 TI - Myosin-Va binds to and mechanochemically couples microtubules to actin filaments. AB - Myosin-Va was identified as a microtubule binding protein by cosedimentation analysis in the presence of microtubules. Native myosin-Va purified from chick brain, as well as the expressed globular tail domain of this myosin, but not head domain bound to microtubule-associated protein-free microtubules. Binding of myosin-Va to microtubules was saturable and of moderately high affinity (approximately 1:24 Myosin-Va:tubulin; Kd = 70 nM). Myosin-Va may bind to microtubules via its tail domain because microtubule-bound myosin-Va retained the ability to bind actin filaments resulting in the formation of cross-linked gels of microtubules and actin, as assessed by fluorescence and electron microscopy. In low Ca2+, ATP addition induced dissolution of these gels, but not release of myosin-Va from MTs. However, in 10 microM Ca2+, ATP addition resulted in the contraction of the gels into aster-like arrays. These results demonstrate that myosin-Va is a microtubule binding protein that cross-links and mechanochemically couples microtubules to actin filaments. PMID- 14565973 TI - Capacity of the golgi apparatus for biogenesis from the endoplasmic reticulum. AB - It is unclear whether the mammalian Golgi apparatus can form de novo from the ER or whether it requires a preassembled Golgi matrix. As a test, we assayed Golgi reassembly after forced redistribution of Golgi matrix proteins into the ER. Two conditions were used. In one, ER redistribution was achieved using a combination of brefeldin A (BFA) to cause Golgi collapse and H89 to block ER export. Unlike brefeldin A alone, which leaves matrix proteins in relatively large remnant structures outside the ER, the addition of H89 to BFA-treated cells caused ER accumulation of all Golgi markers tested. In the other, clofibrate treatment induced ER redistribution of matrix and nonmatrix proteins. Significantly, Golgi reassembly after either treatment was robust, implying that the Golgi has the capacity to form de novo from the ER. Furthermore, matrix proteins reemerged from the ER with faster ER exit rates. This, together with the sensitivity of BFA remnants to ER export blockade, suggests that presence of matrix proteins in BFA remnants is due to cycling via the ER and preferential ER export rather than their stable assembly in a matrix outside the ER. In summary, the Golgi apparatus appears capable of efficient self-assembly. PMID- 14565974 TI - The eps8 family of proteins links growth factor stimulation to actin reorganization generating functional redundancy in the Ras/Rac pathway. AB - Sos-1, a guanine nucleotide exchange factor (GEF), eps8 and Abi1, two signaling proteins, and the lipid kinase phosphoinositide 3-kinase (PI3-K), assemble in a multimolecular complex required for Rac activation leading to actin cytoskeletal remodeling. Consistently, eps8 -/- fibroblasts fail to form membrane ruffles in response to growth factor stimulation. Surprisingly, eps8 null mice are healthy, fertile, and display no overt phenotype, suggesting the existence of functional redundancy within this pathway. Here, we describe the identification and characterization of a family of eps8-related proteins, comprising three novel gene products, named eps8L1, eps8L2, and eps8L3. Eps8Ls display collinear topology and 27-42% identity to eps8. Similarly to eps8, eps8Ls interact with Abi1 and Sos-1; however, only eps8L1 and eps8L2 activate the Rac-GEF activity of Sos-1, and bind to actin in vivo. Consistently, eps8L1 and eps8L2, but not eps8L3, localize to PDGF-induced, F-actin-rich ruffles and restore receptor tyrosine kinase (RTK)-mediated actin remodeling when expressed in eps8 -/- fibroblasts. Thus, the eps8Ls define a novel family of proteins responsible for functional redundancy in the RTK-activated signaling pathway leading to actin remodeling. Finally, the patterns of expression of eps8 and eps8L2 in mice are remarkably overlapping, thus providing a likely explanation for the lack of overt phenotype in eps8 null mice. PMID- 14565976 TI - The Caenorhabditis elegans microtubule-severing complex MEI-1/MEI-2 katanin interacts differently with two superficially redundant beta-tubulin isotypes. AB - The microtubule-severing protein complex katanin is required for a variety of important microtubule-base morphological changes in both animals and plants. Caenorhabditis elegans katanin is encoded by the mei-1 and mei-2 genes and is required for oocyte meiotic spindle formation and must be inactivated before the first mitotic cleavage. We identified a mutation, sb26, in the tbb-2 beta-tubulin gene that partially inhibits MEI-1/MEI-2 activity: sb26 rescues lethality caused by ectopic MEI-1/MEI-2 expression during mitosis, and sb26 increases meiotic defects in a genetic background where MEI-1/MEI-2 activity is lower than normal. sb26 does not interfere with MEI-1/MEI-2 microtubule localization, suggesting that this mutation likely interferes with severing. Tubulin deletion alleles and RNA-mediated interference revealed that TBB-2 and the other germline enriched beta-tubulin isotype, TBB-1, are redundant for embryonic viability. However, limiting MEI-1/MEI-2 activity in these experiments revealed that MEI-1/MEI-2 preferentially interacts with TBB-2-containing microtubules. Our results demonstrate that these two superficially redundant beta-tubulin isotypes have functionally distinct roles in vivo. PMID- 14565975 TI - Interactions between centromere complexes in Saccharomyces cerevisiae. AB - We have purified two new complexes from Saccharomyces cerevisiae, one containing the centromere component Mtw1p together with Nnf1p, Nsl1p, and Dsn1p, which we call the Mtw1p complex, and the other containing Spc105p and Ydr532p, which we call the Spc105p complex. Further purifications using Dsn1p tagged with protein A show, in addition to the other components of the Mtw1p complex, the two components of the Spc105p complex and the four components of the previously described Ndc80p complex, suggesting that all three complexes are closely associated. Fluorescence microscopy and immunoelectron microscopy show that Nnf1p, Nsl1p, Dsn1p, Spc105p, and Ydr532p all localize to the nuclear side of the spindle pole body and along short spindles. Chromatin immunoprecipitation assays show that all five proteins are associated with centromere DNA. Homologues of Nsl1p and Spc105p in Schizosaccharomyces pombe also localize to the centromere. Temperature-sensitive mutations of Nsl1p, Dsn1p, and Spc105p all cause defects in chromosome segregation. Synthetic-lethal interactions are found between temperature-sensitive mutations in proteins from all three complexes, in agreement with their close physical association. These results show an increasingly complex structure for the S. cerevisiae centromere and a probable conservation of structure between parts of the centromeres of S. cerevisiae and S. pombe. PMID- 14565977 TI - ARNO and ARF6 regulate axonal elongation and branching through downstream activation of phosphatidylinositol 4-phosphate 5-kinase alpha. AB - In the developing nervous system, controlled neurite extension and branching are critical for the establishment of connections between neurons and their targets. Although much is known about the regulation of axonal development, many of the molecular events that regulate axonal extension remain unknown. ADP-ribosylation factor nucleotide-binding site opener (ARNO) and ADP-ribosylation factor (ARF)6 have important roles in the regulation of the cytoskeleton as well as membrane trafficking. To investigate the role of these molecules in axonogenesis, we expressed ARNO and ARF6 in cultured rat hippocampal neurons. Expression of catalytically inactive ARNO or dominant negative ARF6 resulted in enhanced axonal extension and branching and this effect was abrogated by coexpression of constitutively active ARF6. We sought to identify the downstream effectors of ARF6 during neurite extension by coexpressing phosphatidyl-inositol-4-phosphate 5 Kinase alpha [PI(4)P 5-Kinase alpha] with catalytically inactive ARNO and dominant negative ARF6. We found that PI(4)P 5-Kinase alpha plays a role in neurite extension and branching downstream of ARF6. Also, expression of inactive ARNO/ARF6 depleted the actin binding protein mammalian ena (Mena) from the growth cone leading edge, indicating that these effects on axonogenesis may be mediated by changes in cytoskeletal dynamics. These results suggest that ARNO and ARF6, through PI(4)P 5-Kinase alpha, regulate axonal elongation and branching during neuronal development. PMID- 14565978 TI - The dynamic association of RCC1 with chromatin is modulated by Ran-dependent nuclear transport. AB - Regulator of chromosome condensation (RCC1) binding to chromatin is highly dynamic, as determined by fluorescence recovery after photobleaching analysis of GFP-RCC1 in stably transfected tsBN2 cells. Microinjection of wild-type or Q69L Ran markedly slowed the mobility of GFP-RCC1, whereas T24N Ran (defective in nucleotide loading) decreased it further still. We found significant alterations in the mobility of intranuclear GFP-RCC1 after treatment with agents that disrupt different Ran-dependent nuclear export pathways. Leptomycin B, which inhibits Crm1/RanGTP-dependent nuclear export, significantly increased the mobility of RCC1 as did high levels of actinomycin D (to inhibit RNA polymerases I, II, and III) or alpha-amanitin (to inhibit RNA polymerases II and III) as well as energy depletion. Inhibition of just mRNA transcription, however, had no affect on GFP RCC1 mobility consistent with mRNA export being a Ran-independent process. In permeabilized cells, cytosol and GTP were required for the efficient release of GFP-RCC1 from chromatin. Recombinant Ran would not substitute for cytosol, and high levels of supplemental Ran inhibited the cytosol-stimulated release. Thus, RCC1 release from chromatin in vitro requires a factor(s) distinct from, or in addition to, Ran and seems linked in vivo to the availability of Ran-dependent transport cargo. PMID- 14565979 TI - Rap1 affects the length and heterogeneity of human telomeres. AB - Telomere length is controlled in part by cis-acting negative regulators that limit telomere extension by telomerase. In budding yeast, the major telomere length regulator scRap1 binds to telomeric DNA and acts to inhibit telomere elongation in cis. Because the human Rap1 ortholog hRap1 does not bind to telomeric DNA directly but is recruited to telomeres by TRF2, we examined its role in telomere length control. The data are consistent with hRap1 being a negative regulator of telomere length, indicating functional conservation. Deletion mapping confirmed that hRap1 is tethered to telomeres through interaction of its C terminus with TRF2. The telomere length phenotypes of hRap1 deletion mutants implicated both the BRCT and Myb domain as protein interaction domains involved in telomere length regulation. By contrast, scRap1 binds to telomeres with its Myb domains and uses its C terminus to recruit the telomere length regulators Rif1 and Rif2. Together, our data show that although the role of Rap1 at telomeres has been largely conserved, the domains of Rap1 have undergone extensive functional changes during eukaryotic evolution. Surprisingly, hRap1 alleles lacking the BRCT domain diminished the heterogeneity of human telomeres, indicating that hRap1 also plays a role in the regulation of telomere length distribution. PMID- 14565980 TI - Induction of S. cerevisiae filamentous differentiation by slowed DNA synthesis involves Mec1, Rad53 and Swe1 checkpoint proteins. AB - A key question in eukaryotic differentiation is whether there are common regulators or biochemical events that are required for diverse types of differentiation or whether there is a core mechanism for differentiation. The unicellular model organism Saccharomyces cerevisiae undergoes filamentous differentiation in response to environmental cues. Because conserved cell cycle regulators, the mitotic cyclin-dependent kinase Clb2/Cdc28, and its inhibitor Swe1 were found to be involved in both nitrogen starvation- and short chain alcohol-induced filamentous differentiation, they were identified as components of the core mechanism for filamentous differentiation. We report here that slowed DNA synthesis also induces yeast filamentous differentiation through conserved checkpoint proteins Mec1 and Rad53. Swe1 and Clb2 are also involved in this form of differentiation, and the core status of Swe1/Clb2/Cdc28 in the mechanism of filamentous differentiation has therefore been confirmed. Because the cAMP and filamentous growth mitogen-activated protein kinase pathways that mediate nitrogen starvation-induced filamentous differentiation are not required for slowed DNA synthesis-induced filamentous growth, they can therefore be excluded from the core mechanism. More significantly, slowed DNA synthesis also induces differentiation in mammalian cancer cells, and such stimulus conservation may indicate that the core mechanism for yeast filamentous differentiation is conserved in mammalian differentiation. PMID- 14565982 TI - The adhesin Hwp1 and the first daughter cell localize to the a/a portion of the conjugation bridge during Candida albicans mating. AB - The cell wall protein Hwp1 was originally demonstrated to be expressed exclusively in hyphae of Candida albicans and cross-linked to human epithelium by mammalian transglutaminase. Hwp1 is expressed on the walls of hyphae formed by a/alpha, a/a, and alpha/alpha cells. Hence, it is expressed on hyphae independently of mating type. However, Hwp1 is selectively expressed on the wall of conjugation tubes formed by a/a cells, but not alpha/alpha cells, in the mating process. This was demonstrated in all possible crosses between four unrelated natural a/a strains and four unrelated alpha/alpha strains. In zygotes, Hwp1 is restricted to that portion of the wall of the conjugation bridge contributed by the a/a parent cell. Hwp1 staining further revealed that the first daughter bud that emerges from the conjugation bridge does so from the a/a contributed portion. Hwp1 expression and localization during the mating process is, therefore, mating type specific, opaque phase specific, and alpha-pheromone induced. These results indicate that the mating type-specific contributions to the conjugation bridge during the mating process in C. albicans are qualitatively and functionally distinct and that the a/a portion of the bridge, which selectively contains Hwp1, bears the first daughter cell in the mating process. PMID- 14565981 TI - Components of U3 snoRNA-containing complexes shuttle between nuclei and the cytoplasm and differentially localize in nucleoli: implications for assembly and function. AB - U3 small nucleolar RNA (snoRNA) and associated proteins are required for the processing of preribosomal RNA (pre-rRNA) and assembly of preribosomes. There are two major U3 snoRNA-containing complexes. The monoparticle contains U3 snoRNA and the core Box C/D snoRNA-associated proteins and an early preribosome-associated complex contains the monoparticle and additional factors that we refer to as preribosome-associated proteins. To address how and where the U3 snoRNA containing preribosome assembles and how these processes are temporally and spatially regulated, we have examined the dynamics and distribution of human U3 complex-associated components in cells with active or inactive transcription of rDNA. We found that U3 complex-associated proteins shuttle between the nucleus and the cytoplasm independent of the synthesis and export of preribosomal particles, suggesting that the shuttling of these proteins may either provide opportunities for their regulation, or contribute to or modulate ribosome export. In addition, monoparticle and preribosome associated components predominantly localize to different nucleolar substructures, fibrillar components, and granular components, respectively, in active nucleoli, and partition separately into the two components during nucleolar segregation induced by inhibition of pol I transcription. Although the predominant localizations of these two sets of factors differ, there are significant areas of overlap that may represent the sites where they reside as a single complex. These results are consistent with a model in which U3 monoparticles associate with the fibrillar components of nucleoli and bind pre-rRNA during transcription, triggering recruitment of preribosome-associated proteins to assemble the complex necessary for pre-rRNA processing. PMID- 14565983 TI - Dissection of the Ascaris sperm motility machinery identifies key proteins involved in major sperm protein-based amoeboid locomotion. AB - Although Ascaris sperm motility closely resembles that seen in many other types of crawling cells, the lamellipodial dynamics that drive movement result from modulation of a cytoskeleton based on the major sperm protein (MSP) rather than actin. The dynamics of the Ascaris sperm cytoskeleton can be studied in a cell free in vitro system based on the movement of plasma membrane vesicles by fibers constructed from bundles of MSP filaments. In addition to ATP, MSP, and a plasma membrane protein, reconstitution of MSP motility in this cell-free extract requires cytosolic proteins that orchestrate the site-specific assembly and bundling of MSP filaments that generates locomotion. Here, we identify a fraction of cytosol that is comprised of a small number of proteins but contains all of the soluble components required to assemble fibers. We have purified two of these proteins, designated MSP fiber proteins (MFPs) 1 and 2 and demonstrated by immunolabeling that both are located in the MSP cytoskeleton in cells and in fibers. These proteins had reciprocal effects on fiber assembly in vitro: MFP1 decreased the rate of fiber growth, whereas MFP2 increased the growth rate. PMID- 14565984 TI - Lipid-dependent bidirectional traffic of apolipoprotein B in polarized enterocytes. AB - Enterocytes are highly polarized cells that transfer nutrients across the intestinal epithelium from the apical to the basolateral pole. Apolipoprotein B (apoB) is a secretory protein that plays a key role in the transepithelial transport of dietary fatty acids as triacylglycerol. The evaluation of the control of apoB traffic by lipids is therefore of particular interest. To get a dynamic insight into this process, we used the enterocytic Caco-2 cells cultured on microporous filters, a system in which the apical and basal compartments can be delimited. Combining biochemical and morphological approaches, our results showed that, besides their role in protection from degradation, lipids control the intracellular traffic of apoB in enterocytes. A supply of fatty acids and cholesterol is sufficient for the export of apoB from the endoplasmic reticulum and its post-Golgi traffic up to the apical brush-border domain, where it remains until an apical supply of complex lipid micelles signals its chase down to the basolateral secretory domain. This downward traffic of apoB involves a microtubule-dependent process. Our results demonstrate an enterocyte-specific bidirectional process for the lipid-dependent traffic of a secretory protein. PMID- 14565985 TI - Characterization of a Drosophila centrosome protein CP309 that shares homology with Kendrin and CG-NAP. AB - The centrosome in animal cells provides a major microtubule-nucleating site that regulates the microtubule cytoskeleton temporally and spatially throughout the cell cycle. We report the identification in Drosophila melanogaster of a large coiled-coil centrosome protein that can bind to calmodulin. Biochemical studies reveal that this novel Drosophila centrosome protein, centrosome protein of 309 kDa (CP309), cofractionates with the gamma-tubulin ring complex and the centrosome-complementing activity. We show that CP309 is required for microtubule nucleation mediated by centrosomes and that it interacts with the gamma-tubulin small complex. These findings suggest that the microtubule-nucleating activity of the centrosome requires the function of CP309. PMID- 14565986 TI - Analysis of the dynein-dynactin interaction in vitro and in vivo. AB - Cytoplasmic dynein and dynactin are megadalton-sized multisubunit molecules that function together as a cytoskeletal motor. In the present study, we explore the mechanism of dynein-dynactin binding in vitro and then extend our findings to an in vivo context. Solution binding assays were used to define binding domains in the dynein intermediate chain (IC) and dynactin p150Glued subunit. Transient overexpression of a series of fragments of the dynein IC was used to determine the importance of this subunit for dynein function in mammalian tissue culture cells. Our results suggest that a functional dynein-dynactin interaction is required for proper microtubule organization and for the transport and localization of centrosomal components and endomembrane compartments. The dynein IC fragments have different effects on endomembrane localization, suggesting that different endomembranes may bind dynein via distinct mechanisms. PMID- 14565987 TI - Platelet-activating factor: a previously unrecognized mediator of fever. AB - Lipopolysaccharide (LPS)-induced systemic inflammation is accompanied by either hypothermia (prevails when the ambient temperature (Ta) is subneutral) or fever (prevails when Ta is neutral or higher). Because platelet-activating factor (PAF) is a proximal mediator of LPS inflammation, it should mediate both thermoregulatory responses to LPS. That PAF possesses hypothermic activity and mediates LPS-induced hypothermia is known. We asked whether PAF possesses pyrogenic activity (Expt 1) and mediates LPS fever (Expt 2). The study was conducted in Long-Evans rats implanted with jugular catheters. A complex with bovine serum albumin (BSA) was infused as a physiologically relevant form of PAF; free (aggregated) PAF was used as a control. In Expt 1, either form of PAF caused hypothermia when infused (83 pmol kg-1 min-1, 60 min, i.v.) at a subneutral Ta of 20 degrees C, but the response to the PAF-BSA complex (-4.5 +/- 0.5 degrees C, nadir) was ~4 times larger than that to free PAF. At a neutral Ta of 30 degrees C, both forms caused fever preceded by tail skin vasoconstriction, but the febrile response to PAF-BSA (1.0 +/- 0.1 degrees C, peak) was > 2 times higher than that to free PAF. Both the hypothermic (at 20 degrees C) and febrile (at 30 degrees C) responses to PAF-BSA started when the total amount of PAF infused was extremely small, < 830 pmol kg-1. In Expt 2 (conducted at 30 degrees C), the PAF receptor antagonist BN 52021 (29 micromol kg-1, i.v.) had no thermal effect of itself. However, it strongly (~2 times) attenuated the febrile response to PAF (5 nmol kg-1, i.v.), implying that this response involves the PAF receptor and is not due to a detergent-like effect of PAF on cell membranes. BN 52021 (but not its vehicle) was similarly effective in attenuating LPS (10 microg kg-1, i.v.) fever. It is concluded that PAF is a highly potent endogenous pyrogenic substance and a mediator of LPS fever. PMID- 14565989 TI - Exercise increases Ca2+-calmodulin-dependent protein kinase II activity in human skeletal muscle. AB - There is evidence in rodents that Ca2+-calmodulin-dependent protein kinase II (CaMKII) activity is higher in contracting skeletal muscle, and this kinase may regulate skeletal muscle function and metabolism during exercise. To investigate the effect of exercise on CaMKII in human skeletal muscle, healthy men (n = 8) performed cycle ergometer exercise for 40 min at 76 +/- 1% peak pulmonary O2 uptake (VO2peak), with skeletal muscle samples taken at rest and after 5 and 40 min of exercise. CaMKII expression and activities were examined by immunoblotting and in vitro kinase assays, respectively. There were no differences in maximal (+ Ca2+, CaM) CaMKII activity during exercise compared with rest. Autonomous (- Ca2+, CaM) CaMKII activity was 9 +/- 1% of maximal at rest, remained unchanged at 5 min, and increased to 17 +/- 1% (P < 0.01) at 40 min. CaMKII autophosphorylation at Thr287 was 50-70% higher during exercise, with no differences in CaMKII expression. The effect of maximal aerobic exercise on CaMKII was also examined (n = 9), with 0.7- to 1.5-fold increases in autonomous CaMKII activity, but no change in maximal CaMKII activity. CaMKIV was not detected in human skeletal muscle. In summary, exercise increases the activity of CaMKII in skeletal muscle, suggesting that it may have a role in regulating skeletal muscle function and metabolism during exercise in humans. PMID- 14565990 TI - A circadian clock in the fish retina regulates dopamine release via activation of melatonin receptors. AB - Although many biochemical, morphological and physiological processes in the vertebrate retina are controlled by a circadian (24 h) clock, the location of the clock and how the clock alters retinal function are unclear. For instance, several observations have suggested that dopamine, a retinal neuromodulator, may play an important role in retinal rhythmicity but the link between dopamine and a clock located within or outside the retina remains to be established. We found that endogenous dopamine release from isolated goldfish retinae cultured in continuous darkness for 56 h clearly exhibited a circadian rhythm with high values during the subjective day. The continuous presence of melatonin (1 nM) in the culture medium abolished the circadian rhythm of dopamine release and kept values constantly low and equal to the night-time values. The selective melatonin antagonist luzindole (1 microM) also abolished the dopamine rhythm but the values were high and equal to the daytime values. Melatonin application during the late subjective day introduced rod input and reduced cone input to fish cone horizontal cells, a state usually observed during the subjective night. In contrast, luzindole application during the subjective night decreased rod input and increased cone input. Prior application of dopamine or spiperone, a selective dopamine D(2)-like antagonist, blocked the above effects of melatonin and luzindole, respectively. These findings indicate that a circadian clock in the vertebrate retina regulates dopamine release by the activation of melatonin receptors and that endogenous melatonin modulates rod and cone pathways through dopamine-mediated D(2)-like receptor activation. PMID- 14565991 TI - Pacing-induced calcineurin activation controls cardiac Ca2+ signalling and gene expression. AB - Calcineurin, a Ca(2+)-calmodulin-dependent protein phosphatase (PP2B) is one of the links between Ca(2+) signals and regulation of gene transcription in cardiac muscle. We studied the Ca(2+) signal specificity of calcineurin activation experimentally and with modelling. In the rat atrial preparation, an increase in pacing frequency increased nuclear activity of the calcineurin-sensitive transcription factor, nuclear factor of activated T-cells (NFAT), 2-fold in a cyclosporin A (CsA)-sensitive manner. In line with this, modelling results predicted that the frequency of cardiac Ca(2+) transients encodes the stimulus for calcineurin activation. We further observed experimentally that calcineurin inhibition by CsA modulated Ca(2+) release in a Ca(2+)-dependent manner. CsA had no effect on [Ca(2+)](i) at a pacing frequency of 1 Hz but it significantly suppressed the amplitude of Ca(2+) transients, systolic [Ca(2+)](i) and time averaged [Ca(2+)](i) at 6 Hz. Calcineurin had a differential role in the expression of immediate-early genes B-type natriuretic peptide (BNP) and c-fos. CsA inhibited the pacing-induced BNP gene expression, whereas pacing alone had no effect on the expression of c-fos. However, in the presence of CsA, c-fos mRNA levels were significantly augmented by increased pacing frequency. These results show that frequency-dependent calcineurin activation has a specific role in [Ca(2+)](i) regulation and gene expression, constantly recruited by varying cardiac Ca(2+) signals. PMID- 14565992 TI - Alternative splicing of N- and C-termini of a C. elegans ClC channel alters gating and sensitivity to external Cl- and H+. AB - CLH-3 is a meiotic cell cycle-regulated ClC Cl- channel that is functionally expressed in oocytes of the nematode Caenorhabditis elegans. CLH-3a and CLH-3b are alternatively spliced variants that have identical intramembrane regions, but which exhibit striking differences in their N- and C-termini. Structural and functional studies indicate that N- and C-terminal domains modulate ClC channel activity. We therefore postulated that alternative splicing of CLH-3 would alter channel gating and physiological functions. To begin testing this hypothesis, we characterized the biophysical properties of CLH-3a and CLH-3b expressed heterologously in HEK293 cells. CLH-3a activates more slowly and requires stronger hyperpolarization for activation than CLH-3b. Depolarizing conditioning voltages dramatically increase CLH-3a current amplitude and induce a slow inactivation process at hyperpolarized voltages, but have no significant effect on CLH-3b activity. CLH-3a also differs significantly in its extracellular Cl- and pH sensitivity compared to CLH-3b. Immunofluorescence microscopy demonstrated that CLH-3b is translationally expressed during all stages of oocyte development, and furthermore, the biophysical properties of the native oocyte Cl- current are indistinguishable from those of heterologously expressed CLH-3b. We conclude that CLH-3b carries the oocyte Cl- current and that the channel probably functions in nonexcitable cells to depolarize membrane potential and/or mediate net Cl- transport. The unique voltage-dependent properties of CLH-3a suggest that the channel may function in muscle cells and neurones to regulate membrane excitability. We suggest that alternative splicing of CLH-3 N- and C-termini modifies the functional properties of the channel by altering the accessibility and/or function of pore-associated ion-binding sites. PMID- 14565995 TI - Properties of pacemaker potentials recorded from myenteric interstitial cells of Cajal distributed in the mouse small intestine. AB - Recording of electrical responses from isolated small intestine of mice using conventional microelectrodes revealed two types of potential, a pacemaker potential and a slow wave, both with rapid rising primary components and following plateau components. The rate of rise and peak amplitude were greater for pacemaker potentials than for slow waves, and the plateau component was smaller in slow waves than in pacemaker potentials. Both potentials oscillated at a similar frequency (20-30 min-1). Unitary potentials often discharged during the interval between pacemaker potentials. Infusion of Lucifer Yellow allowed visualization of the recorded cells; pacemaker potentials were recorded from myenteric interstitial cells of Cajal (ICC-MY) while slow waves were recorded from circular smooth muscle cells. Pacemaker potentials were characterized as follows: the primary component was inhibited by Ni2+, Ca2+-free solution or depolarization with high-K+ solution, the plateau component was inhibited by 4,4' diisothiocyanostilbene-2,2'-disulphonic acid (DIDS), an inhibitor of Ca2+ activated Cl- channels, low [Cl-]o solution or Ca2+-free solution, and the generation of potentials was abolished by co-application of Ni2+and DIDS or by chelating intracellular Ca2+ with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N' tetraacetic acid acetoxymethyl ester (BAPTA-AM). These results indicate that in the mouse small intestine ICC-MY generate pacemaker potentials with two components in situ; the primary and plateau components may be generated by activation of voltage-dependent Ca2+-permeable channels and Ca2+-activated Cl- channels, respectively. Slow waves are generated in circular smooth muscles via electrotonic spread of pacemaker potentials. These properties of intestinal pacemaker potentials are considered essentially similar to those of gastric pacemaker potentials. PMID- 14565994 TI - The effect of recombinant human growth hormone and resistance training on IGF-I mRNA expression in the muscles of elderly men. AB - The expression of two isoforms of insulin-like growth factor-I (IGF-I): mechano growth factor (MGF) and IGF-IEa were studied in muscle in response to growth hormone (GH) administration with and without resistance training in healthy elderly men. A third isoform, IGF-IEb was also investigated in response to resistance training only. The subjects (age 74 +/- 1 years, mean +/- S.E.M) were assigned to either resistance training with placebo, resistance training combined with GH administration or GH administration alone. Real-time quantitative RT-PCR was used to determine mRNA levels in biopsies from the vastus lateralis muscle at baseline, after 5 and 12 weeks in the three groups. GH administration did not change MGF mRNA at 5 weeks, but significantly increased IGF-IEa mRNA (237%). After 12 weeks, MGF mRNA was significantly increased (80%) compared to baseline. Five weeks of resistance training significantly increased the mRNA expression of MGF (163%), IGF-IEa (68%) and IGF-IEb (75%). No further changes were observed after 12 weeks. However, after 5 weeks of training combined with GH treatment, MGF mRNA increased significantly (456%) and IGF-IEa mRNA by (167%). No further significant changes were noted at 12 weeks. The data suggest that when mechanical loading in the form of resistance training is combined with GH, MGF mRNA levels are enhanced. This may reflect an overall up-regulation of transcription of the IGF-I gene prior to splicing. PMID- 14566000 TI - Hyperosmolar solution effects in guinea pig airways. II. Epithelial bioelectric responses to relative changes in osmolarity. AB - Osmotic challenge of airways alters the bioelectric properties of the airway epithelium and induces the release of factors that modulate smooth muscle tone. Recent studies in our laboratory suggested that methacholine-contracted airways relax in response to incremental increases in osmolarity, rather than from cell shrinkage or absolute solute concentration. In the present study, guinea pig tracheae were mounted in Ussing chambers to elucidate the bioelectric effects of challenge of the epithelium with hyperosmolar and isosmolar solutions. Transepithelial short-circuit current (Isc) across tracheae stimulated with basolateral methacholine was inhibited by apical amiloride, apical 5-nitro-2-(3 phenylpropylamino)benzoic acid, basolateral bumetanide, basolateral ouabain, and Cl(-)-free solution, but not by basolateral iberiotoxin. Apical hyperosmolar challenge with NaCl variably decreased or increased Isc, but D-mannitol (D-M) always inhibited Isc; bumetanide attenuated decreases in Isc. The effects of the transport blockers depended upon whether Isc was initially decreased or increased. Unique concentration-dependent changes in Isc and transepithelial resistance (Rt) were observed when ionic (NaCl and KCl), nonionic impermeant (D-M and sucrose), and nonionic permeant (urea) osmolytes were added to the apical and basolateral baths. At concentrations that doubled the osmolarity of the apical bath, D-M, urea, and N-methyl-D-glucamine-gluconate (NMDG-Glu) decreased Isc. Apical isosmolar NMDG-Glu solution decreased Isc, and additional NMDG-Glu caused a further decrease in Isc. Inclusion of one permeant ion, either Na+,K+, or Cl-, reversed the response to apical isosmolar and hyperosmolar solutions. Thus, bioelectric responses of the airway epithelium to hyperosmolar solution are induced by incremental increases in osmolarity. PMID- 14566001 TI - Hyperosmolar solution effects in guinea pig airways. III. Studies on the identity of epithelium-derived relaxing factor in isolated perfused trachea using pharmacological agents. AB - Hyperosmolar challenge of airway epithelium stimulates the release of epithelium derived relaxing factor (EpDRF), but the identity of EpDRF is not known. We examined the effects of pharmacological agents on relaxant responses of methacholine (3 x 10(-7) M)-contracted guinea pig perfused trachea to mucosal hyperosmolar challenge using D-mannitol. Responses were inhibited by gossypol (5 x 10(-6) M), an agent with diverse actions, by the carbon monoxide (CO) scavenger hemoglobin (10(-6) M), and by the heme oxygenase (HO) inhibitor zinc (II) protoporphyrin IX (10(-4) M). The HO inhibitor chromium (III) mesoporphyrin IX (10(-4) M) was not inhibitory, and the HO activator heme-L-lysinate (3 x 10(-4) M) did not evoke relaxant responses. The CO donor tricarbonyldichlororuthenium (II) dimer (2.2 x 10(-4) M) elicited small relaxation responses. Other agents without an effect on responses included: apyrase, adenosine, 6-anilino-5,8 quinolinequinone (LY83583), proadifen, (E)-3-[[[3-[2-(7-chloro-2 quinolinyl)ethenyl]phenyl][[3-(dimethylamino)-3-oxopropyl]thio]methyl]thio] propanoic acid (MK 571), diphenhydramine, glibenclamide, HgCl2, tetrodotoxin, nystatin, alpha-hemolysin, 8-bromoguanosine 3',5'-cyclic monophosphothioate, Rp isomer, 12-O-tetradecanoylphorbol-13-acetate, cholera toxin, pertussis toxin, thapsigargin, nifedipine, Ca(2+)-free mucosal solution, hydrocortisone, and epidermal growth factor. Cytoskeleton inhibitors, includingerythro-9-(2-hydroxyl 3-nonyl)adenine, colchicine, nocodazole, latrunculin B, and cytochalasins B and D, had no effect on relaxation responses. The results suggest provisionally that a portion of EpDRF activity may be due to CO and that the release of EpDRF does not involve cytoskeletal reorganization. PMID- 14566002 TI - Hyperosmolar solution effects in guinea pig airways. IV. Lipopolysaccharide induced alterations in airway reactivity and epithelial bioelectric responses to methacholine and hyperosmolarity. AB - We investigated the in vivo and in vitro effects of lipopolysaccharide (LPS) treatment (4 mg/kg i.p.) on guinea pig airway smooth muscle reactivity and epithelial bioelectric responses to methacholine (MCh) and hyperosmolarity. Hyperosmolar challenge of the epithelium releases epithelium-derived relaxing factor (EpDRF). Using a two-chamber, whole body plethysmograph 18 h post treatment, animals treated with LPS were hyporeactive to inhaled MCh aerosol. This could involve an increase in the release and/or actions of EpDRF, because LPS treatment enhanced EpDRF-induced smooth muscle relaxation in vitro in the isolated perfused trachea apparatus. In isolated perfused tracheas the basal transepithelial potential difference (Vt) was increased after LPS treatment. The increase in Vt was inhibited by amiloride and indomethacin. Concentration response curves for changes in Vt in response to serosally and mucosally applied MCh were biphasic (hyperpolarization, <3 x 10(-7)M; depolarization, >3 x 10( 7)M); MCh was more potent when applied serosally. The hyperpolarization response to MCh, but not the depolarization response, was potentiated after LPS treatment. In both treatment groups, mucosally applied hyperosmolar solution (using added NaCl) depolarized the epithelium; this response was greater in tracheas from LPS treated animals. The results of this study indicate that airway hyporeactivity in vivo after LPS treatment is accompanied by an increase in the release and/or actions of EpDRF in vitro. These changes may involve LPS-induced bioelectric alterations in the epithelium. PMID- 14565999 TI - Actomyosin energy turnover declines while force remains constant during isometric muscle contraction. AB - Energy turnover was measured during isometric contractions of intact and Triton permeabilized white fibres from dogfish (Scyliorhinus canicula) at 12 degrees C. Heat + work from actomyosin in intact fibres was determined from the dependence of heat + work output on filament overlap. Inorganic phosphate (Pi) release by permeabilized fibres was recorded using the fluorescent protein MDCC-PBP, N-(2-[1 maleimidyl]ethyl)-7-diethylamino-coumarin-3 carboxamide phosphate binding protein. The steady-state ADP release rate was measured using a linked enzyme assay. The rates decreased five-fold during contraction in both intact and permeabilized fibres. In intact fibres the rate of heat + work output by actomyosin decreased from 134 +/-s.e.m. 28 microW mg(-1) (n = 17) at 0.055 s to 42% of this value at 0.25 s, and to 20% at 3.5 s. The force remained constant between 0.25 and 3.5 s. Similarly in permeabilized fibres the Pi release rate decreased from 5.00 +/- 0.39 mmol l(-1) s(-1) at 0.055 s to 39% of this value at 0.25 s and to 19% at 0.5 s. The steady-state ADP release rate at 15 s was 21% of the Pi rate at 0.055 s. Using a single set of rate constants, the time courses of force, heat + work and Pi release were described by an actomyosin model that took account of the transition from the initial state (rest or rigor) to the contracting state, shortening and the consequent work against series elasticity, and reaction heats. The model suggests that increasing Pi concentration slows the cycle in intact fibres, and that changes in ATP and ADP slow the cycle in permeabilized fibres. PMID- 14566003 TI - Extended in vivo pharmacodynamic activity of E5564 in normal volunteers with experimental endotoxemia [corrected]. AB - E5564 (alpha-D-glucopyranose) is a synthetic antagonist of bacterial endotoxin that has been shown to completely block human endotoxin response. Low doses of E5564 (0.35-3.5 mg) have a long pharmacokinetic half-life, but a surprisingly short ex vivo and in vivo pharmacodynamic half-life (generally less than several hours). To determine whether extended antagonistic activity can be achieved in vivo, this study assesses the pharmacodynamic activity of 4- and 72-h infusions of E5564 into normal volunteers. Administration of 3.5 mg of E5564/h x 72 h completely blocked effects of endotoxin challenge at the end of dosing (72 h), and at 48 and 72 h postdosing. Similarly, a 4-h infusion of E5564, 3 mg/h completely blocked endotoxin administered 8 h postdosing. A lower dose of E5564, 0.5 mg/h x 4 h, ameliorated but did not block most effects of endotoxin 8 h postdosing (p <0.05). Finally, the effect of varying plasma lipoprotein content on E5564 activity was studied in subjects having high or low cholesterol levels (>180 or <140 mg/dl) after 72-h infusion of 252 mg of E5564. No differences were observed. These results demonstrate that E5564 blocks the effects of endotoxin in a human model of clinical sepsis and indicate its potential in the treatment and/or prevention of clinical sepsis. PMID- 14566004 TI - Saxitoxin blocks L-type ICa. AB - Saxitoxin (STX) and tetrodotoxin (TTX) are frequently used to selectively block sodium channels. In this study, we provide evidence that commercial STX also inhibits L-type Ca2+ currents (I(Ca,L)) in adult mouse ventricular myocytes (VMs) and tsA-201 cells that were transiently cotransfected with three calcium channel subunits. We measured inhibition of sodium currents (INa) in mouse VMs, of I(Ca,L) in mouse VM and tsA-201 cells, and intracellular calcium concentration ([Ca2+]i) transients in single mouse VMs. STX or TTX was abruptly applied before the test voltage pulse using a rapid solution switcher device. STX (10 microM; Calbiochem) and TTX (60 microM; Sigma-Aldrich) completely blocked INa in mouse VMs. However, STX at 10 microM also reduced I(Ca,L) in mouse VM by 39% (P < 0.0001; n = 14), whereas TTX at 60 microM had no effect on I(Ca,L). STX (10 microM; Calbiochem) reduced the amplitude of the [Ca2+]i transients in mouse VMs by 36% (P < 0.0001; n = 10). In contrast, TTX (60 microM; Sigma-Aldrich) only reduced the amplitude of the [Ca2+]i transients by 9% (P = 0.003; n = 5). STX (10 microM) obtained from Sigma-Aldrich showed a similar inhibitory effect on I(Ca,L) (33%) (P < 0.0001; n = 5) in mouse VMs. STX (Calbiochem) inhibited the calcium currents of tsA-201 cells in a dose-dependent manner. This inhibition was voltage independent. The current-voltage relationship of calcium currents in tsA-201 cells was not altered by STX. These results indicate that STX partially blocks L type Ca2+ channels and thus provide further evidence that its effects are not specific for Na+ channels. PMID- 14566005 TI - Ion-trapping, microsomal binding, and unbound drug distribution in the hepatic retention of basic drugs. AB - This study investigated the relative contribution of ion-trapping, microsomal binding, and distribution of unbound drug as determinants in the hepatic retention of basic drugs in the isolated perfused rat liver. The ionophore monensin was used to abolish the vesicular proton gradient and thus allow an estimation of ion-trapping by acidic hepatic vesicles of cationic drugs. In vitro microsomal studies were used to independently estimate microsomal binding and metabolism. Hepatic vesicular ion-trapping, intrinsic elimination clearance, permeability-surface area product, and intracellular binding were derived using a physiologically based pharmacokinetic model. Modeling showed that the ion trapping was significantly lower after monensin treatment for atenolol and propranolol, but not for antipyrine. However, no changes induced by monensin treatment were observed in intrinsic clearance, permeability, or binding for the three model drugs. Monensin did not affect binding or metabolic activity in vitro for the drugs. The observed ion-trapping was similar to theoretical values estimated using the pHs and fractional volumes of the acidic vesicles and the pKa values of drugs. Lipophilicity and pKa determined hepatic drug retention: a drug with low pKa and low lipophilicity (e.g., antipyrine) distributes as unbound drug, a drug with high pKa and low lipophilicity (e.g., atenolol) by ion trapping, and a drug with a high pKa and high lipophilicity (e.g., propranolol) is retained by ion-trapping and intracellular binding. In conclusion, monensin inhibits the ion-trapping of high pKa basic drugs, leading to a reduction in hepatic retention but with no effect on hepatic drug extraction. PMID- 14566006 TI - Identification of a potent and selective noncovalent cathepsin S inhibitor. AB - Cathepsin S is considered crucial for normal presentation of major histocompatibility complex (MHC) class II-restricted antigens by antigen presenting cells to CD4+ T cells. It is a key enzyme for the degradation of the class II-associated invariant chain, a process that is required for effective antigen loading of class II molecules. Here, we report a selective, orally available, high-affinity cathepsin S inhibitor, 1-[3-[4-(6-Chloro-2,3-dihydro-3 methyl-2-oxo-1H-benzimidazol-1-yl)-1-piperidinyl]propyl]-4,5,6,7-tetrahydro-5 (methylsulfonyl)-3-[4-(trifluoromethyl)phenyl]-1H-pyrazolo[4,3-c]pyridine. (JNJ 10329670), that represents a novel class of immunosuppressive compounds. JNJ 10329670 is a highly potent (Ki of approximately 30 nM), nonpeptidic, noncovalent inhibitor of human cathepsin S, but it is much less active against the mouse, dog, monkey, and bovine enzymes. The compound is inactive against other proteases, including the closely related cathepsins L, F, and K. This selectivity makes JNJ 10329670 an excellent tool for exploring the role of cathepsin S in human systems. Treatment of human B cell lines and primary human dendritic cells with JNJ 10329670 resulted in the accumulation of the p10 fragment of the invariant chain (IC50 of approximately 1 microM). In contrast, inhibition of invariant chain proteolysis was much less effective in a human monocytic cell line, suggesting that other enzymes may degrade the invariant chain in this cell type. JNJ 10329670 was shown to block the proteolysis of the invariant chain in vivo by using immunocompromised mice injected with human peripheral blood mononuclear cells (PBMCs). Furthermore, this inhibitor blocks the presentation of tetanus toxoid and giant ragweed by human PBMCs. The properties of JNJ 10329670 make it a candidate for immunosuppressive therapy of allergies and autoimmune diseases. PMID- 14566007 TI - An electrophysiological model of spinal transmission deficits in mouse experimental autoimmune encephalomyelitis. AB - Chronic relapsing/remitting experimental autoimmune encephalomyelitis (EAE) can be induced in 8-week-old female SJL/J(H-2) mice via inoculation with the p139-151 peptide of myelin proteolipid protein (PLP), Mycobacterium tuberculosis (MT), complete Freund's adjuvant (CFA), and Bordatella pertussis. EAE is a relevant preclinical model of MS that incorporates several aspects of the clinical disease. Chief among these are the inflammatory mediated neurological deficits. While the impact of localized spinal cord demyelination on neurotransmission has been modeled successfully, relatively little work has been done with spinal cord from animals with EAE. The goal of this study was to assess the utility of a grease-gap tissue bath methodology in the detection of transmission deficits in EAE spinal cord tissue. Spinal cords removed from EAE mice at different phases of the neurological deficit were assessed for their response to both lumbar and sacral application of one of several depolarizing agents (veratridine, potassium chloride [KCl], (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid [AMPA]). The main finding of this study is that transmission deficits were detected in EAE mice at the onset of the neurological deficits. They were sustained for a period of approximately 2 to 3 weeks post disease onset followed by a gradual recovery of group function. The other finding is that there is a decrease in the latency to achieve AMPA-mediated depolarization in sacral spinal cord that is independent of the magnitude of the depolarization response. These results suggest that this methodology can be utilized to assess sensory and motor deficits in spinal cord from EAE animals. PMID- 14566008 TI - Sensitization of neuronal A2A adenosine receptors after persistent D2 dopamine receptor activation. AB - Acute activation of Galpha(i/o)-coupled D2 dopamine receptors inhibits A2A adenosine receptor stimulation of adenylate cyclase. This antagonistic interaction between D2 dopamine and A2A adenosine receptors has been well documented; however, the effects of persistent activation of D2 dopamine receptors on subsequent A2A adenosine receptor signaling have not been explored. The present study investigated the effects of short-term (3-h) and long-term (18 h) activation of D2L dopamine receptors on subsequent A2A adenosine receptor stimulation of adenylate cyclase in CAD-D2L and NS20Y-D2L neuroblastoma cells. Short- and long-term activation of D2L dopamine receptors markedly increased 5'-N methylcarboxamidoadenosine (MECA)-stimulated cyclic AMP accumulation 1.4-fold and 1.7-fold, respectively. D2L receptor-induced sensitization of A2A-stimulated cyclic AMP accumulation was blocked by the D2 antagonist spiperone and pertussis toxin pretreatment. In addition, persistent activation of A2A adenosine receptors resulted in 50% desensitization of subsequent MECA-stimulated cyclic AMP accumulation; however, MECA-induced desensitization of A2A adenosine receptors did not prevent completely quinpirole-induced sensitization of adenylate cyclase. These studies revealed a novel mode of regulation between D2L dopamine and A2A adenosine receptors and suggest a cooperative interaction in the regulation of cyclic AMP signaling. PMID- 14566009 TI - Synthetic high-density lipoproteins exert cardioprotective effects in myocardial ischemia/reperfusion injury. AB - Human high-density lipoproteins (HDLs) protect the heart against ischemia/reperfusion injury. In the present study, the cardioprotective effects of synthetic high-density lipoproteins (sHDLs) made of phosphatidylcholine and apolipoprotein A-I were investigated in isolated rat hearts, which underwent a 20 min low-flow ischemia followed by a 30-min reperfusion. The administration of sHDL during the 10 min immediately before ischemia caused a rapid, dose-dependent improvement of postischemic cardiac function: at the maximum dose (2.0 mg of sHDL protein/ml), left ventricular developed pressure (LVDP) recovered to 71.0 +/- 3.2 versus 40.5 +/- 3.8 mm Hg in saline-treated hearts, and coronary perfusion pressure (CPP) increased to 100.3 +/- 6.2 versus 132.0 +/- 9.0 mm Hg. The preservation of postischemic cardiac function was associated with a dose dependent reduction of creatine kinase release into the coronary effluent. sHDLs administered in the first 10 min postischemia also exerted a significant, dose dependent improvement of postischemic LVDP, CPP, and creatine kinase release, but the cardioprotective effect was less than when sHDLs were given preischemia. The preservation of postischemic cardiac function by sHDL was mediated through a reduction of cardiac tumor necrosis factor-alpha content and an enhanced cardiac production of prostaglandin E2 and I2. The present experimental data indicate that sHDLs may provide a novel therapeutic approach to clinical conditions in which myocardial ischemia/reperfusion occurs, such as acute coronary syndromes, cardiac surgery, or revascularization procedures. PMID- 14566010 TI - Suppression of cytochrome P450 2E1 promoter activity by interferon-gamma and loss of response due to the -71G>T nucleotide polymorphism of the CYP2E1*7B allele. AB - The CYP2E1*7B allele is defined by two nucleotide sequence polymorphisms, -71G>T and -333T>A. The CYP2E1 promoter sequence flanking the -71G nucleotide is consistent with a gamma-interferon activated sequence. Inflammation and interferon (IFN)-gamma suppress expression of CYP2E1 in vivo; however, the exact mechanism is not known. The objectives of this study were to determine whether the CYP2E1 promoter is regulated by IFN-gamma and to examine the influence of the nucleotide substitutions on this function. Treatment of HepG2 cells with IFN gamma, after transient transfection with a luciferase reporter gene bearing the native CYP2E1 (-71G) promoter sequence resulted, in a dose-dependent reduction of luciferase activity. In contrast, no suppression was observed in cells transfected with the *7B allele promoter (-333A and -71T) nor a CYP2E1 plasmid containing only the -71T polymorphism. These data indicate that IFN-gamma suppresses native CYP2E1 promoter activity and that the -71G is critical for this response. PMID- 14566011 TI - Ovarian steroid hormone modulation of the acute effects of cocaine on luteinizing hormone and prolactin levels in ovariectomized rhesus monkeys. AB - Cocaine stimulates significant increases in luteinizing hormone (LH) and decreases prolactin levels in gonadally intact rhesus monkeys, but cocaine did not alter plasma levels of these anterior pituitary hormones in ovariectomized females. These findings suggested that ovarian steroid hormones may contribute to the endocrine effects of acute cocaine administration. To test this hypothesis, the acute effects of cocaine and placebo-cocaine on plasma LH and prolactin levels were examined in five ovariectomized rhesus females during three chronic hormone replacement conditions: 1) estradiol (E2beta) treatment (0.0015-0.006 mg/kg/day i.m.), 2) progesterone treatment (0.32 mg/kg/day i.m.), and 3) combinations of progesterone (0.32 mg/kg/day i.m.) and E2beta (0.002 and 0.004 mg/kg/day i.m.). Cocaine (0.8 mg/kg i.v.) did not alter prolactin or LH in ovariectomized monkeys without ovarian steroid replacement. During chronic estradiol treatment, cocaine produced an estradiol dose-dependent decrease in prolactin. Cocaine also decreased prolactin during treatment with progesterone alone and progesterone + E2beta (0.004 mg/kg/day i.m.). Cocaine stimulated a significant increase in LH during treatment with progesterone alone, but not during treatment with progesterone + E2beta, or three of four estradiol treatment doses. Cocaine pharmacokinetics did not differ as a function of hormone replacement conditions. Together, these data suggest that both E2beta and progesterone modulate cocaine's effects on prolactin, whereas E2beta alone and in combination with progesterone, do not facilitate LH release in response to cocaine in ovariectomized rhesus females. PMID- 14566012 TI - Cardiovascular effects of nicotine, chlorisondamine, and mecamylamine in the pigeon. AB - Chlorisondamine and mecamylamine are nicotinic antagonists that produce both ganglionic and central blockade. Chlorisondamine, when administered as a large systemic dose, produces a persistent central block, despite being charged. The present study evaluated the cardiovascular effects of chlorisondamine. Shortly after administration, chlorisondamine (0.10, 1, and 10 mg/kg i.m.) lowered blood pressure significantly and decreased heart rate at the low dose (0.1 mg/kg i.m.) and increased heart rate at the high dose (10 mg/kg i.m.). Mecamylamine (1 and 10 mg/kg i.m.) also lowered blood pressure and heart rate. After both antagonists, heart rate returned to baseline values within 90 min and blood pressure within 24 h. Low doses of nicotine (0.01-0.03 mg/kg i.m.) lowered blood pressure but did not affect heart rate. Higher doses (0.10-3.2 mg/kg i.m.) transiently increased blood pressure and heart rate. Subsequent to antagonist administration, nicotine was administered to determine whether either drug blocked the cardiovascular effects of nicotine. Chlorisondamine (0.1, 1, and 10 mg/kg i.m.) administered 30 min before nicotine blocked the increases in blood pressure and heart rate. Only the high dose (10 mg/kg i.m.) of chlorisondamine administered 24 h before nicotine produced a blockade of nicotine's pressor effect. This block diminished within 3 days. Mecamylamine (1 mg/kg i.m.) antagonized only nicotine's tachycardic effect. Longer pretreatment with mecamylamine (10 mg/kg, 24 h before nicotine challenge) did not antagonize the cardiovascular effects of nicotine. Thus, chlorisondamine produces a longer lasting blockade of nicotine's cardiovascular effects than mecamylamine. PMID- 14566014 TI - Three-dimensional organization of pKi-67: a comparative fluorescence and electron tomography study using FluoroNanogold. AB - The monoclonal antibody (MAb) Ki-67 is routinely used in clinical studies to estimate the growth fraction of tumors. However, the role of pKi-67, the protein detected by the Ki-67 MAb, remains elusive, although some biochemical data strongly suggest that it might organize chromatin. To better understand the functional organization of pKi-67, we studied its three-dimensional distribution in interphase cells by confocal microscopy and electron tomography. FluoroNanogold, a single probe combining a dense marker with a fluorescent dye, was used to investigate pKi-67 organization at the optical and ultrastructural levels. Observation by confocal microscopy followed by 3D reconstruction showed that pKi-67 forms a shell around the nucleoli. Double labeling experiments revealed that pKi-67 co-localizes with perinucleolar heterochromatin. Electron microscopy studies confirmed this close association and demonstrated that pKi-67 is located neither in the fibrillar nor in the granular components of the nucleolus. Finally, spatial analyses by electron tomography showed that pKi-67 forms cords 250-300 nm in diameter, which are themselves composed of 30-50-nm thick fibers. These detailed comparative in situ analyses strongly suggest the involvement of pKi-67 in the higher-order organization of perinucleolar chromatin. PMID- 14566015 TI - Sex-related expression of 20alpha-hydroxysteroid dehydrogenase mRNA in the adult mouse. AB - The enzyme 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD) catalyzes the conversion of progesterone into its inactive form, 20alpha-hydroxyprogesterone. To gain information about the exact sites of 20alpha-HSD mRNA expression, we performed in situ hybridization using a (35)S-labeled cRNA probe in tissues of adult mice of both sexes. 20alpha-HSD mRNA was expressed in both male and female gonads. In the ovary, high expression was found in luteal cells of corpora lutea, while much lower expression could be detected in granulosa cells of growing follicles. In the testis, a specific hybridization signal was detected only in Leydig cells. In the female reproductive tract, 20alpha-HSD mRNA was found in the epithelial cells of the uterine cervix. In the adrenal cortex, only the zona reticularis exhibited specific radiolabeling, the expression being very high in the female and very low in the male. In the skin, specific labeling was restricted to sebaceous glands, the hybridization signal being much higher in the female than in the male. In the liver, 20alpha-HSD mRNA was found in hepatocytes, with a higher degree of expression in the female. In the kidney, specific labeling was observed in the epithelial cells of distal convoluted tubules, the signal being also much more striking in the female than in the male. In non reproductive tissues, it clearly appears that the expression of 20alpha-HSD mRNA is higher in the female than in the male, suggesting that 20alpha-HSD may play an important role in reducing the intracellular concentration of progesterone originating from the circulation at a much higher level in the female. PMID- 14566016 TI - C-Met expression and mechanical activation of satellite cells on cultured muscle fibers. AB - Single-fiber cultures can be used to model satellite cell activation in vivo. Although technical deficiencies previously prevented study of stretch-induced events, here we describe a method developed to study satellite cell gene expression by in situ hybridization (ISH) using protocol modifications for fiber adhesion and fixation. The hypothesis that mechanical stretching activates satellite cells was tested. Fiber cultures were established from normal flexor digitorum brevis muscles and plated on FlexCell dishes with a layer of Vitrogen. After 2 hr of stretch in the presence of BrdU, satellite cells on fibers attached to Vitrogen were activated above control levels. In the absence of activating treatments or mechanical stretch, ISH studies showed 0-6 c-Met+ satellite cells per fiber. Time course experiments demonstrated stable quiescence in the absence of stretch and significant peaks in activation after 30 min and 2 hr of stretch. Frequency distributions for unstretched fiber cultures showed a significantly greater number of quiescent c-Met+ satellite cells than were activated by stretching, suggesting that typical activation stimuli did not trigger cycling in the entire c-Met+ population of satellite cells. These methods have a strong potential to further dissect the nature of stretch-induced activation and gene expression among characterized populations of individual quiescent and activated satellite cells. PMID- 14566013 TI - Perlecan and tumor angiogenesis. AB - Perlecan is a major heparan sulfate proteoglycan (HSPG) of basement membranes (BMs) and connective tissues. The core protein of perlecan is divided into five domains based on sequence homology to other known proteins. Commonly, the N terminal domain I of mammalian perlecan is substituted with three HS chains that can bind a number of matrix molecules, cytokines, and growth factors. Perlecan is essential for metazoan life, as shown by genetic manipulations of nematodes, insects, and mice. There are also known human mutations that can be lethal. In vertebrates, new functions of perlecan emerged with the acquisition of a closed vascular system and skeletal connective tissues. Many of perlecan's functions may be related to the binding and presentation of growth factors to high-affinity tyrosine kinase (TK) receptors. Data are accumulating, as discussed here, that similar growth factor-mediated processes may have unwanted promoting effects on tumor cell proliferation and tumor angiogenesis. Understanding of these attributes at the molecular level may offer opportunities for therapeutic intervention. PMID- 14566017 TI - Follicular expression of c-Kit/SCF and inhibin-alpha in mouse ovary during development. AB - The mechanism of development of the ovarian follicles has been largely unknown. We performed an immunohistochemical (IHC) study to determine the follicular expressions of c-kit, SCF, and inhibin-alpha at different developmental stages in mouse ovary. Ovaries were obtained from 14 and 16 days post coitum and 2, 7, and 21 days post partum (dpp) mice. IHC for c-kit, SCF, and inhibin-alpha was carried out. c-Kit and SCF were expressed on oogonia regardless of the developmental stage. Immunoreactive c-kit and SCF antigens were expressed on oocytes of primordial and primary follicles of neonate mouse ovaries. In 21 dpp mouse ovary, the expression of c-kit/SCF in oocytes gradually decreased as the follicles developed. c-Kit/SCF was expressed strongly in oocytes of preantral follicles and weakly in granulosa and thecal cells. Inhibin-alpha was mainly expressed on granulosa cells of preantral and early antral follicles of the 21 dpp mouse ovaries. These findings suggest that the IHC expression of c-kit/SCF proteins is specific in all developmental stages of ovarian follicles and is decreased after the follicle starts to grow. The expression of inhibin-alpha is negatively correlated with the expression of c-kit/SCF in the ovarian follicles in mice. PMID- 14566018 TI - Expression and localization of prohormone convertase PC1 in the calcitonin producing cells of the bullfrog ultimobranchial gland. AB - We examined the expression and localization of the prohormone convertases, PC1 and PC2, in the ultimobranchial gland of the adult bullfrog using immunohistochemical (IHC) and in situ hybridization (ISH) techniques. In the ultimobranchial gland, PC1-immunoreactive cells were columnar, and were present in the follicular epithelium. When serial sections were immunostained with anti calcitonin, anti-CGRP, anti-PC1, and anti-PC2 sera, PC1 was found only in the calcitonin/CGRP-producing cells. No PC2-immunopositive cells were detected. In the ISH, PC1 mRNA-positive cells were detected in the follicle cells in the ultimobranchial gland. No PC2 mRNA-positive cells were detected. RT-PCR revealed expression of the mRNAs of PC1 and the PC2 in the ultimobranchial gland. However, very little of the PC2 mRNA is probably translated because no PC2 protein was detected either by IHC staining or by Western blotting analysis. We conclude that the main prohormone convertase that is involved in the proteolytic cleavage of procalcitonin in the bullfrog is PC1. PMID- 14566019 TI - Morphofunctional studies of the glomerular wall in mice lacking entactin-1. AB - The architecture of the basement membranes is essential for proper function. This architecture is based on interactions among its components, which assemble in a complex network. Entactin-1 appears to be the mastermind of this assembling. In entactin-1-null transgenic mice, immunocytochemistry established the absence of entactin-1 in the glomerular basement membrane, and morphological thickening of this membrane was demonstrated. This prompted us to investigate the organization of other components of the glomerular basement membrane in the transgenic animals. The distribution of type IV collagen and laminin remained unchanged, whereas that of anionic charges was significantly altered. We also evaluated the impact of the absence of entactin-1 on cell relays by studying the alpha(3)- and the alpha(v)-integrins along the endothelial and epithelial glomerular cell plasma membranes. Only the density of alpha(v) was found to be increased. Finally, the filtration properties of the glomerular wall were evaluated by revealing endogenous albumin distribution across the basement membrane. This was altered in transgenic animals, suggesting changes in permselectivity properties. Entactin-1 appears to be an essential component in basement membranes because its absence appears to modify the molecular organization leading to alterations in functional properties. PMID- 14566020 TI - Expression of Myosin heavy chain isoforms in rat soleus muscle spindles after 19 days of hypergravity. AB - The aim of this study was to determine whether a period of 19 days in hypergravity was long enough to induce changes in the expression of myosin heavy chain (MyHC) isoforms in the muscle spindles. The soleus muscle of 10 male Wistar rats (control: CONT, n=5; hypergravity: HG, n=5) was frozen, cut into serial sections, and labeled with antibodies against MyHCs: I, IIA, IIA + IIX + IIB, slow-tonic, and alpha-cardiac. Forty CONT and 45 HG spindles were analyzed. The results from HG spindles compared to CONT showed that there was no change in the cross-sectional area of intrafusal fibers. However, along the entire length of B1 fibers, the expression of both MyHC I and alpha-cardiac was increased significantly, whereas the labeling against MyHC IIA and MyHC slow-tonic was decreased. In B2 fibers, the labeling against MyHC IIA (region A), slow-tonic (region A), and fast myosins (regions A-C) was statistically decreased. In chain fibers, the labeling against both MyHC IIA and fast MyHC was reduced significantly. We conclude that hypergravity has a real impact on the MyHC content in the muscle spindles and induces some inverse changes of those observed in hypogravity for MyHCs I, alpha-cardiac, and slow-tonic. PMID- 14566021 TI - Network organization of interstitial connective tissue cells in the human endolymphatic duct. AB - The human endolymphatic duct (ED) and sac of the inner ear have been suggested to control endolymph volume and pressure. However, the physiological mechanisms for these processes remain obscure. We investigated the organization of the periductal interstitial connective tissue cells and extracellular matrix (ECM) in four freshly fixed human EDs by transmission electron microscopy and by immunohistochemistry. The unique surgical material allowed a greatly improved structural and epitopic preservation of tissue. Periductal connective tissue cells formed frequent intercellular contacts and focally occurring electron-dense contacts to ECM structures, creating a complex tissue network. The connective tissue cells also formed contacts with the basal lamina of the ED epithelium and the bone matrix, connecting the ED with the surrounding bone of the vestibular aqueduct. The interstitial connective tissue cells were non-endothelial and non smooth muscle fibroblastoid cells. We suggest that the ED tissue network forms a functional mechanical entity that takes part in the control of inner ear fluid pressure and endolymph resorption. PMID- 14566022 TI - Localization of cholecystokinin receptor subtypes in the endocine pancreas. AB - This study was undertaken to clarify the controversy in the literature about pancreatic localization of the cholecystokinin (CCK) CCK(A) and CCK(B) receptors. With antibodies used by other investigators, we first established their specificity by Western blotting, indirect immunofluorescence, and confocal microscopy with each antibody's peptide antigen. Co-localization assays between the CCK receptors and the pancreatic hormones insulin, glucagon, and somatostatin revealed that the CCK(A) RAbs 1122 and R1-2 recognized insulin and glucagon cells in rat, pig, and human pancreas but not in the somatostatin cells. Conversely, the three CCK(B) RAbs tested, 9262, 9491, and GR4, identified the somatostatin cells. Abs 9491 and GR4 occasionally co-localized with glucagon, a feature that never occurred with Ab 9262. Finally, the specificity of Ab 9262 for the pancreatic CCK(B) R was confirmed in six different species. It co-localized with somatostatin but never with glucagon in these species. Our data suggest the use of Abs 1122 and 9262 to specifically identify and localize pancreatic CCK(A) and CCK(B) receptors, respectively. Confusion in the literature may result from the lack of specificity of most antibodies used, as established in this study. PMID- 14566023 TI - Cellular distribution of NDRG1 protein in the rat kidney and brain during normal postnatal development. AB - N-myc downregulated gene 1 (NDRG1) is a 43-kD protein whose mRNA is induced by DNA damage, hypoxia, or prolonged elevation of intracellular calcium. Although NDRG1 is also upregulated during cell differentiation, there are few studies on NDRG1 expression during postnatal development. Here we investigated the expression and cellular distribution of NDRG1 protein in rat kidney and brain during postnatal development. Immunohistochemical analysis revealed that the cellular localization of NDRG1 protein in the kidney changed from the proximal convoluted tubules to the collecting ducts between postnatal days 10 and 20. In the brain, a change in cellular expression was also found from the hippocampal pyramidal neurons to the astrocytes in the gray matter during the same postnatal period. These alterations in the cellular distribution of NDRG1 were associated with shifts in the molecular assembly on Western blots. Under non-reduced conditions, the main NDRG1 band was found only around 215 kD in both kidney and brain during the early postnatal stage. After postnatal day 10, the immunoreactive bands shifted to 43 kD in the kidney and 129 kD in the brain. These changes in the cellular distribution and state of assembly may correlate with the functional maturation of both organs. PMID- 14566024 TI - Cytokeratin immunoreactivity in lobular intraepithelial neoplasia. AB - Eighteen commercially available antibodies reactive against different cytokeratin proteins were tested on classic examples of lobular intraepithelial neoplasia (LIN) and of ductal intraepithelial neoplasia (DIN) of the breast. About 90% of higher-grade DIN (AIDH and DCIS) show no or substantially diminished reaction with clone 34betaE12 (specified as reactive against keratins 1, 5, 10, and 14 as determined by the manufacturer), while the cells of LIN were found to express the antigen reactive with this antibody. To determine which of these four keratins are present in the cells of LIN, antibodies reactive against these individual four keratins were tested. None of the four antibodies to keratins 1, 5, 10, or 14 reacted with the cells of LIN. To investigate this further, 13 additional monoclonal antibodies to various other keratin proteins were tested on the cells of LIN. Those that successfully reacted with the cells of LIN were further tested on the cells of DIN. All of the individual antibodies reactive with the cells of LIN were also reactive with the cells of DIN to a degree, with clone RCK108 (reactive against keratin 19) coming the closest to demonstrating the reactivity seen with 34betaE12. We conclude that the reactivity seen in the cells of LIN with 34betaE12 is due to either (a) a crossreaction with keratin 19 that is slightly less prominent than the reaction of the individual clone RCK108, (b) a crossreaction with a keratin protein that was not tested (3, 11, 12), (c) a crossreaction with a protein closely resembling keratin in formalin-fixed, paraffin-embedded tissue, or (d) the detection of a mutated or truncated form of keratin 1, 5, 10, or 14 that cannot be detected by the individual monoclonal antibody. PMID- 14566025 TI - Vimentin-positive cells in the epithelium of rabbit ileal villi represent cup cells but not M-cells. AB - Membranous (M)-cells are specialized epithelial cells of the Peyer's patch domes that transport antigens from the intestinal lumen to the lymphoid tissue. Vimentin is a reliable marker for M-cells in rabbits. Using immunohistochemistry (IHC), a subpopulation of epithelial cells has recently been identified in ordinary rabbit ileal villi, which are vimentin-positive and share morphological characteristics with the M-cells of the domes. To test the hypothesis that these cells represent M-cells outside the organized lymphoid tissue, lectin labeling and tracer uptake experiments were performed. Lectins specific for N-acetyl glucosamine oligomers selectively bound to the vimentin-positive villous cells but not to M-cells in the domes. Microbeads instilled into the ileal lumen were taken up by M-cells within 45 min but not by the vimentin-positive cells in the villi. Lectin-gold labeling on ultrathin sections revealed that the lectin binding sites were located in the brush border and in vesicles in the apical cytoplasm. The vimentin/lectin-positive cells shared ultrastructural characteristics with the so-called "cup cells." We conclude (a) that the vimentin positive cells in ordinary villi represent cup cells but not M-cells, (b) that they are readily detectable by (GlucNAc)(N)-specific lectins, and (c) that they do not transcytose experimental tracers. Although the specific function of cup cells is still obscure, they most probably represent a cell type distinct from M cells of the domes with respect to both function and expression of the two new markers. PMID- 14566026 TI - Cell-specific expression of CYP2A5 in the mouse respiratory tract: effects of olfactory toxicants. AB - We performed a detailed analysis of mouse cytochrome P450 2A5 (CYP2A5) expression by in situ hybridization (ISH) and immunohistochemistry (IHC) in the respiratory tissues of mice. The CYP2A5 mRNA and the corresponding protein co-localized at most sites and were predominantly detected in the olfactory region, with an expression in sustentacular cells, Bowman's gland, and duct cells. In the respiratory and transitional epithelium there was no or only weak expression. The nasolacrimal duct and the excretory ducts of nasal and salivary glands displayed expression, whereas no expression occurred in the acini. There was decreasing expression along the epithelial linings of the trachea and lower respiratory tract, whereas no expression occurred in the alveoli. The hepatic CYP2A5 inducers pyrazole and phenobarbital neither changed the CYP2A5 expression pattern nor damaged the olfactory mucosa. In contrast, the olfactory toxicants dichlobenil and methimazole induced characteristic changes. The damaged Bowman's glands displayed no expression, whereas the damaged epithelium expressed the enzyme. The CYP2A5 expression pattern is in accordance with previously reported localization of protein and DNA adducts and the toxicity of some CYP2A5 substrates. This suggests that CYP2A5 is an important determinant for the susceptibility of the nasal and respiratory epithelia to protoxicants and procarcinogens. PMID- 14566027 TI - Expression of hedgehog proteins in the human thymus. AB - The Hedgehog (Hh) family of secreted proteins includes intercellular signaling molecules that specify cell fate and patterning during the development of many tissues. In this study we show that the different components of the Hh signaling pathway are expressed in human thymus. The three mammalian Hh proteins, Sonic (Shh), Indian (Ihh), and Desert (Dhh) hedgehog, are produced by thymic epithelial cells. Shh-expressing epithelial cells are restricted to the thymic subcapsula and medulla, whereas Ihh- and Dhh-producing epithelial cells are distributed throughout the thymus. The requisite Hh receptors, Patched 1(Ptc1) and Smoothened (Smo), and the Gli transcription factors are expressed by thymocytes and also by epithelial cells. Ptc1 is expressed in most thymocyte subsets, whereas Smo expression is mainly associated with immature thymocytes. The isoform of the Ptc receptor, Ptc2, is expressed only by intrathymic progenitor cells and epithelial cells. Other Hh-binding proteins with modulating functions, such as Hedgehog interacting protein (Hip) and growth arrest-specific gene-1 (Gas-1), are also expressed in human thymus. Our study shows that the intrathymic expression pattern of the Hh signaling pathway components is complex and suggests that Hh proteins may regulate human thymocyte differentiation from the earliest developmental stages, as well as thymic epithelial cell function. PMID- 14566028 TI - Immunocytochemical detection of GLUT2 at the rat intestinal brush-border membrane. AB - We have proposed a new model of intestinal sugar absorption in which high sugar concentrations promote rapid insertion of the facilitative transporter GLUT2 into the brush-border membrane so that absorptive capacity is precisely regulated to match dietary intake during the assimilation of a meal. However, location of GLUT2 at the brush border by immunocytochemistry has been problematical. We report that control of rapid GLUT2 trafficking and the use of an antibody to a sequence within the large extracellular loop of GLUT2 permits localization of GLUT2 at the brush border. To reveal brush-border GLUT2 fully, it is necessary to digest the sugar chain at the glycosylation site close to the antigenic site. In this way, we have demonstrated by immunocytochemistry PKC-dependent changes in the regulation of brush-border GLUT2 in rat jejunum that correspond to those seen by Western blotting. The functional and immunocytochemical data are now reconciled. PMID- 14566029 TI - Relevance of thiopurine methyltransferase status in rheumatology patients receiving azathioprine. AB - Azathioprine (AZA) is widely used in the management of rheumatological diseases. Despite its efficacy, AZA can often cause bone marrow suppression, notably leucopenia, which has been recorded in up to 17% of patients taking AZA for rheumatoid arthritis, though this can be considered clinically significant in about 3% overall. Severe myelosuppression, associated with abnormal AZA metabolism, is linked to the thiopurine methyltransferase (TPMT) genetic polymorphism. TPMT status can be assessed prior to AZA treatment by measuring enzyme activity or genotyping techniques. Analysis of recent data suggests that by optimizing the AZA dose on the basis of TPMT status testing (with a substantial reduction in dose for patients homozygous for mutant TPMT alleles), a reduction in drug-induced morbidity and cost savings can be made by avoiding hospitalization and rescue therapy for leucopenic events. In this article we review the pharmacogenetic and clinical implications of the TPMT polymorphism, emphasizing its relevance to rheumatologists managing diseases with AZA. PMID- 14566030 TI - Post-onset inhibition of murine arthritis using combined chemokine antagonist therapy. AB - OBJECTIVE: To investigate the effect of targeting the chemotaxis of monocytes and polymorphonuclear monocytes (PMNs) in situ in MRL-Faslpr arthritis. METHODS: MRL Faslpr mice were injected intradermally with complete Freund's adjuvant and cellular infiltration into the joint was monitored. Once clinical disease developed, the animals received one of three treatments: MCP-1(9-76); MCP-1(9-76) plus Gro-alpha(8-73); or control peptide, MCP-1 Ala. The bimalleolar ankle width was measured for 11 days and histological examination of the joints was then assessed. RESULTS: Cellular infiltration started after the onset of ankle swelling, and increased progressively. The incidence of swelling and the histopathology was reduced after day 6 of treatment in the MCP-1(9-76)-treated mice. Mice treated with the two antagonists MCP-1(9-76) and Gro-alpha(8-73) displayed a further significant reduction in disease parameters. CONCLUSION: Treatment after disease onset with chemotactic antagonists for monocytes and PMNs significantly alleviated both the swelling and the histopathology seen in arthritis, suggesting that chemokine antagonists are an effective anti inflammatory therapy. PMID- 14566031 TI - The relationship between health-related quality of life, pain and coping strategies in juvenile idiopathic arthritis. AB - OBJECTIVES: To investigate the relationship between health-related quality of life (HRQL), experience of pain and pain coping strategies in children with juvenile idiopathic arthritis (JIA). To compare reports describing these variables obtained from children and their parents. METHODS: Participants were 59 children aged 8 to 18 yr with JIA and their parents. Parents and children completed the PedsQL generic core scales and arthritis module, the visual analogue scale of the Varni-Thompson Pediatric Pain Questionnaire, and the Waldron/Varni Pediatric Pain Coping Inventory. Parents rated children's functional disability using the Childhood Health Assessment Questionnaire. RESULTS: Parents reported significantly lower scores (indicating worse HRQL) than children on five of the eight PedsQL scales rating children's HRQL. Parents and children reported a significant negative relationship between pain levels and the PedsQL scores assessing children's physical, emotional and social functioning. They also reported a significant negative relationship between scores on several pain coping scales and scores on the PedsQL scales. However, the pattern of these relationships varied for reports from parents and children. CONCLUSIONS: Pain intensity and pain coping strategies have a significant and independent relationship with several domains that comprise the HRQL of children with JIA. However, parents and children have differing perceptions of the nature of these relationships. The differences emphasize the importance of clinicians obtaining information about children's HRQL, pain levels and pain coping strategies from both parents and children. PMID- 14566032 TI - Does the application of tape influence quadriceps sensorimotor function in knee osteoarthritis? AB - OBJECTIVES: Using additional data from two previously reported studies, the aim was to determine the effects of immediate and short-term continuous (3 weeks) application of knee tape on quadriceps sensorimotor function in individuals with symptomatic knee osteoarthritis (OA). METHODS: A within-subjects study (n = 18) and a randomized controlled trial (n = 87) were performed using community volunteers. Therapeutic tape (patellar realignment and soft-tissue unloading) was compared with control (placebo) tape and no tape. Outcome measures included knee joint position sense, isometric quadriceps strength and electromyographic quadriceps onset during stair descent. RESULTS: Application of therapeutic tape worsened joint position sense (variable error) at a knee angle of 40 degrees (P < 0.01), but did not immediately alter any other sensorimotor parameter. Even after 3 weeks of wearing tape continuously, sensorimotor function was not altered. Furthermore, no differential effect of tape was noted when participants were dichotomized into those with poor and good sensorimotor scores at baseline. CONCLUSIONS: Neither immediate application nor continuous use of tape (for 3 weeks) appears to influence sensorimotor function in people with knee OA. Alterations in sensorimotor function cannot explain the pain-relieving effects of therapeutic tape observed in this population. PMID- 14566033 TI - Temporary cardiac pacing in district general hospitals--sustainable resource or training liability? PMID- 14566034 TI - Hepatocellular damage from non-steroidal anti-inflammatory drugs. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used for the management of rheumatological disorders, and as analgesics and antipyretics. Hepatotoxicity is an uncommon, but potentially lethal complication, which usually occurs within 12 weeks of starting therapy. It can occur with all NSAIDs, but appears to be more common with diclofenac and particularly sulindac. Female patients aged >50 years, with autoimmune disease, and those on other potentially hepatotoxic drugs, appear to be particularly susceptible. Liver function test abnormalities generally settle within 4-6 weeks of stopping the causative drug. However, some patients may develop acute liver failure and successful orthotopic liver transplantation may be undertaken in such patients. Recent in vitro animal studies have shown that the mechanism of diclofenac toxicity relates both to impairment of ATP synthesis by mitochondria, and to production of active metabolites, particularly n,5-dihydroxydiclofenac, which causes direct cytotoxicity. Mitochondrial permeability transition (MPT) has also been shown to be important in diclofenac-induced liver injury, resulting in generation of reactive oxygen species, mitochondrial swelling and oxidation of NADP and protein thiols. Physicians and hepatologists must be vigilant to the hepatotoxic potential of any NSAID, as increased awareness, surveillance and reporting of these events will lead to a better understanding of the risk factors and the pathophysiology of NSAID-related hepatotoxicity. PMID- 14566035 TI - C-reactive protein and cardiovascular disease: new insights from an old molecule. AB - The classical acute-phase protein, C-reactive protein (CRP), is an exquisitely sensitive systemic marker of disease with broad clinical utility for monitoring and differential diagnosis. Inflammation, the key regulator of CRP synthesis, plays a pivotal role in atherothrombotic cardiovascular disease. There is a powerful predictive association between raised serum CRP values and the outcome of acute coronary syndromes, and, remarkably, between even modestly increased CRP production and future atherothrombotic events in otherwise healthy individuals. Baseline CRP values also reflect metabolic states associated with atherothrombotic events. The presence of CRP within most atherosclerotic plaques and all acute myocardial infarction lesions, coupled with binding of CRP to lipoproteins and its capacity for pro-inflammatory complement activation, suggests that CRP may contribute to the pathogenesis and complications of cardiovascular disease. We review the biological properties of CRP, the association between CRP and cardiovascular disease, and the possibility that CRP may be a novel therapeutic target. PMID- 14566037 TI - Endothelial function in HIV-infected patients receiving protease inhibitor therapy: does immune competence affect cardiovascular risk? AB - BACKGROUND: The use of HIV protease inhibitors (PIs) as a component of combination antiretroviral therapy in HIV-infected patients has been associated with dyslipidaemia, but its significance as a risk factor for cardiovascular disease is unclear. Endothelial dysfunction is an early phase of atherogenesis that may be assessed non-invasively with ultrasonography in vivo. AIM: To evaluate vascular function and investigate potential determinants of endothelial dysfunction of the peripheral circulation in PI-treated, HIV-infected men with dyslipidaemia. DESIGN: Observational, case-control study. METHODS: We studied 24 HIV-infected, PI-treated men with dyslipidaemia and 24 normolipidaemic, healthy male controls matched for age and body mass index. Brachial artery endothelial function was studied using high-resolution ultrasound and computerized edge detection software. This non-invasive technique measured post-ischaemic flow mediated dilatation (FMD), and the endothelium-independent vasodilatory response to glyceryl trinitrate (GTN). RESULTS: Within the HIV patient group, FMD was significantly associated with percentage of 'naive' CD4 + 45RA + T cells (p = 0.03), while plasma lipid/lipoprotein and insulin levels, body mass, and smoking status did not correlate with endothelial function. FMD was not significantly different between the study group and the controls. CONCLUSIONS: The atherogenic potential of PI-associated dyslipidaemia may be attenuated in HIV-infected patients with decreased immune competence, reflecting a possible contribution of cell-mediated immune responses to the pathogenesis of atherosclerosis. PMID- 14566036 TI - Prospects for treatment of paraquat-induced lung fibrosis with immunosuppressive drugs and the need for better prediction of outcome: a systematic review. AB - BACKGROUND: Acute paraquat self-poisoning is a significant problem in parts of Asia, the Pacific and the Caribbean. Ingestion of large amounts of paraquat results in rapid death, but smaller doses often cause a delayed lung fibrosis that is usually fatal. Anti-neutrophil ('immunosuppressive') treatment has been recommended to prevent lung fibrosis, but there is no consensus on efficacy. AIM: To review the evidence for the use of immunosuppression in paraquat poisoning, and to identify validated prognostic systems that would allow the use of data from historical control studies and the future identification of patients who might benefit from immunosuppression. DESIGN: Systematic review. METHODS: We searched PubMed, Embase and Cochrane databases for 'paraquat' together with 'poisoning' or 'overdose'. We cross-checked references and contacted experts, and searched on [www.google.com] and [www.yahoo.com] using 'paraquat', 'cyclophosphamide', 'methylprednisolone' and 'prognosis'. RESULTS: We found ten clinical studies of immunosuppression in paraquat poisoning. One was a randomized controlled trial (RCT). Seven used historical controls only; the other two were small (n = 1 and n = 4). Mortality in controls and patients varied markedly between studies. Three of the seven non-RCT controlled studies measured plasma paraquat; analysis using Proudfoot's or Hart's nomograms did not suggest that immunosuppression increased survival in these studies. Of 16 prognostic systems for paraquat poisoning, none has been independently validated in a large cohort. DISCUSSION: The authors of the RCT have performed valuable and difficult research, but their results are hypothesis-forming rather than conclusive; elsewhere, the use of historical controls is problematic. In the absence of a validated prognostic marker, a large RCT of immunosuppression using death as the primary outcome is required. This RCT should also prospectively test and validate the available prognostic methods, so that future patients can be selected for this and other therapies on admission. PMID- 14566038 TI - Intensified treatment of type 2 diabetes--positive effects on blood pressure, but not glycaemic control. AB - BACKGROUND: Since publication of the UK Prospective Diabetes Study (UKPDS) in 1998, there has been a clear evidence base for tight glycaemic (HBA(1c) < 7.0%) and blood pressure (BP < 140/85 mmHg) control. AIM: To determine the effect of UKPDS-based intensified glycaemic and BP targets on the care of type 2 diabetic patients attending a routine diabetes clinic. DESIGN: Two surveys, each of 500 consecutively attending type 2 diabetic patients. METHODS: The first survey was in a 3-month period in 1999, shortly after publication of the UKPDS study. The second was identical, but 2 years later. Glycaemic control (by DCCT-aligned HBA(1c)), BP and treatment details were recorded in both. RESULTS: BP control was significantly improved in the second survey (mean +/- SD systolic BP from 151 +/- 25 to 146 +/- 26 mmHg, p = 0.001; diastolic from 77 +/- 13 to 72 +/- 12 mmHg, p < 0.0001) and the proportion of patients on anti-hypertensive treatment increased from 33% to 60% (p < 0.0001). Mean HbA(1c) however remained unchanged (8.7 +/- 1.8% in 1999 vs. 8.5 +/- 1.8% in 2001), although there was evidence of more intensive treatment patterns, with declining numbers on diet alone and more on oral agents and/or insulin. DISCUSSION: Intensified BP control may be achievable within the confines of routine diabetes care, but achievement of optimal glycaemic targets remains problematic. PMID- 14566039 TI - Pregnancy in renal transplant recipients: the Royal Free Hospital experience. AB - BACKGROUND: For women with end-stage renal failure of child-bearing age, renal transplantation offers a chance to start a family. Pregnancies in renal transplant recipients involve risks for graft and fetus, and need to be carefully managed. AIM: To identify graft, fetal and maternal outcomes in our patients, and compare our results with those of the large national transplant registries. DESIGN: Retrospective case-note review. METHODS: We assessed the outcomes of 48 pregnancies in 24 renal transplant recipients. Obstetric data and renal parameters were examined in 27-30 pregnancies that progressed to delivery. RESULTS: Mean time from transplantation to pregnancy was 6.5 years, with an unfavourable outcome in patients who conceived within 1 year. There was a 41% incidence of fetal growth restriction (FGR), and 33% of infants were small for gestational age. FGR was associated with maternal hypertension, a pre-pregnancy serum creatinine (SCr) >/= 133 micro mol/l (1.5 mg/dl), calcineurin inhibitors and the use of cardioselective beta blockers. Two patients with pre-pregnancy SCr > 200 micro mol/l lost their grafts within 3 years of delivery. A permanent significant decline in graft function occurred in 20%, by 6 months post delivery. DISCUSSION: FGR with SGA infants occurs frequently. Atenolol should be avoided in pregnancy and Metoprolol should not be combined with calcineurin inhibitors. Pregnancy appeared to have a deleterious effect on graft function in patients with SCr > 155 micro mol (1.75 mg/dl). Patients with pre-pregnancy SCr 200 micro mol/l are at greatest risk. PMID- 14566040 TI - Severe acute respiratory syndrome: global initiatives for disease diagnosis. AB - We present a retrospective analysis of the available articles on severe acute respiratory syndrome (SARS) published since the outbreak of the disease. SARS is a new infectious disease caused by a novel coronavirus. Originating in Guangdong, Southern China, at the end of 2002, it has spread to regions all over the world, affecting more than 8000 people. With high morbidity and mortality, SARS is an important respiratory disease which may be encountered world-wide. The causative virus was identified by a WHO-led network of laboratories, which identified the genome sequence and developed the first molecular assays for diagnosis. For the respiratory physician, detecting SARS in its earliest stages, identifying pathways of transmission, and implementing preventive and therapeutic strategies are all important. The WHO and the CDC have published helpful definitions of 'suspected' and 'probable' cases. However, the symptoms of the disease may change, and laboratory tests and definitions are still limited. Even in a situation of no new cases of infection, SARS remains a major respiratory health hazard. As with influenza virus outbreaks, new epidemics may arise at the end of each year. PMID- 14566042 TI - Familial juvenile hyperuricaemic nephropathy. PMID- 14566043 TI - Acute pancreatitis and exacerbation of hepatitis B following reduced dose of prednisolone. PMID- 14566044 TI - Plasma cholecystokinin concentrations are elevated in acute upper gastrointestinal infections. PMID- 14566045 TI - Misplaced. PMID- 14566046 TI - Folk illness and medical models. PMID- 14566048 TI - Helical Levy walks: adjusting searching statistics to resource availability in microzooplankton. AB - The searching trajectories of different animals can be described with a broad class of flight length (lj) distributions with P(lj) = lj-mu. Theoretical studies have shown that changes in these distributions (i.e., different mu values) are key to optimizing the long-term encounter statistics under certain searcher resource scenarios. In particular, they predict the advantage of Levy searching (mu approximately 2) over Brownian motion (mu > or = 3) for low-prey-density scenarios. Here, we present experimental evidence of predicted optimal changes in the flight-time distribution of a predator's walk in response to gradual density changes of its moving prey. Flight times of the dinoflagellate Oxyrrhis marina switched from an exponential to an inverse square power-law distribution when the prey (Rhodomonas sp.) decreased in abundance. Concomitantly, amplitude and frequency of the short-term helical path increased. The specific biological mechanisms involved in these searching behavioral changes are discussed. We suggest that, in a three-dimensional environment, a stronger helical component combined with a Levy walk searching strategy enhances predator's encounter rates. Our results support the idea of universality of the statistical laws in optimal searching processes despite variations in the biological details of the organisms. PMID- 14566047 TI - The development of diabetes in E2f1/E2f2 mutant mice reveals important roles for bone marrow-derived cells in preventing islet cell loss. AB - Our studies of mice deficient for the E2F1 and E2F2 transcription factors have revealed essential roles for these proteins in the cell cycle control of pancreatic exocrine cells and the regulation of pancreatic beta cell maintenance. Pancreatic exocrine cells in E2f1-/-E2f2 mutant mice become increasingly polyploid with age, coinciding with severe exocrine atrophy. Furthermore, mice deficient for both E2F1 and E2F2 develop nonautoimmune, insulin-dependent diabetes with high penetrance. Surprisingly, transplantation of wild-type bone marrow can prevent or rescue diabetes in E2f1-/-E2f2-/-mice. We hypothesize that exocrine degeneration results in a destructive environment for beta cells, which can be alleviated by restoration of the hematopoietic system that is also defective in E2f1-/-E2f2-/-mice The demonstration that beta cell maintenance under conditions of stress is influenced by bone marrow-derived cells may provide important insight into the design of therapies to boost islet mass and function in diabetic patients. PMID- 14566049 TI - Structure of tRNA pseudouridine synthase TruB and its RNA complex: RNA recognition through a combination of rigid docking and induced fit. AB - RNA pseudouridine synthase, TruB, catalyzes pseudouridine formation at U55 in tRNA. This posttranscriptional modification is almost universally conserved and occurs in the T arm of most tRNAs. We determined the crystal structure of Escherichia coli TruB apo enzyme, as well as the structure of Thermotoga maritima TruB in complex with RNA. Comparison of the RNA-free and -bound forms of TruB reveals that this enzyme undergoes significant conformational changes on binding to its substrate. These conformational changes include the ordering of the "thumb loop," which binds right into the RNA hairpin loop, and a 10 degree hinge movement of the C-terminal domain. Along with the result of docking experiments performed on apo TruB, we conclude that TruB recognizes its RNA substrate through a combination of rigid docking and induced fit, with TruB first rigidly binding to its target and then maximizing the interaction by induced fit. PMID- 14566050 TI - Endogenous DNA double-strand breaks: production, fidelity of repair, and induction of cancer. AB - This article extends our previous quantitative analysis of the relationship between the dynamics of the primary structure of DNA and mutagenesis associated with single-strand lesions to an analysis of the production and processing of endogenous double-strand breaks (EDSBs) and to their implications for oncogenesis. We estimate that in normal human cells approximately 1% of single strand lesions are converted to approximately 50 EDSBs per cell per cell cycle. This number is similar to that for EDSBs produced by 1.5-2.0 Gy of sparsely ionizing radiation. Although EDSBs are usually repaired with high fidelity, errors in their repair contribute significantly to the rate of cancer in humans. The doubling dose for induced DSBs is similar to doubling doses for mutation and for the induction of carcinomas by ionizing radiation. We conclude that rates of production of EDSBs and of ensuing spontaneous mitotic recombination events can account for a substantial fraction of the earliest oncogenic events in human carcinomas. PMID- 14566052 TI - Nonlinear elasticity, proteinquakes, and the energy landscapes of functional transitions in proteins. AB - Large-scale motions of biomolecules involve linear elastic deformations along low frequency normal modes, but for function nonlinearity is essential. In addition, unlike macroscopic machines, biological machines can locally break and then reassemble during function. We present a model for global structural transformations, such as allostery, that involve large-scale motion and possible partial unfolding, illustrating the method with the conformational transition of adenylate kinase. Structural deformation between open and closed states occurs via low-frequency modes on separate reactant and product surfaces, switching from one state to the other when energetically favorable. The switching model is the most straightforward anharmonic interpolation, which allows the barrier for a process to be estimated from a linear normal mode calculation, which by itself cannot be used for activated events. Local unfolding, or cracking, occurs in regions where the elastic stress becomes too high during the transition. Cracking leads to a counterintuitive catalytic effect of added denaturant on allosteric enzyme function. It also leads to unusual relationships between equilibrium constant and rate like those seen recently in single-molecule experiments of motor proteins. PMID- 14566051 TI - Mutations in human complement regulator, membrane cofactor protein (CD46), predispose to development of familial hemolytic uremic syndrome. AB - Membrane cofactor protein (MCP; CD46) is a widely expressed transmembrane complement regulator. Like factor H it inhibits complement activation by regulating C3b deposition on targets. Factor H mutations occur in 10-20% of patients with hemolytic uremic syndrome (HUS). We hypothesized that MCP mutations could predispose to HUS, and we sequenced MCP coding exons in affected individuals from 30 families. MCP mutations were detected in affected individuals of three families: a deletion of two amino acids (D237/S238) in family 1 (heterozygous) and a substitution, S206P, in families 2 (heterozygous) and 3 (homozygous). We evaluated protein expression and function in peripheral blood mononuclear cells from these individuals. An individual with the D237/S238 deletion had reduced MCP levels and approximately 50% C3b binding compared with normal controls. Individuals with the S206P change expressed normal quantities of protein, but demonstrated approximately 50% reduction in C3b binding in heterozygotes and complete lack of C3b binding in homozygotes. MCP expression and function was evaluated in transfectants reproducing these mutations. The deletion mutant was retained intracellularly. S206P protein was expressed on the cell surface but had a reduced ability to prevent complement activation, consistent with its reduced C3b binding and cofactor activity. This study presents further evidence that complement dysregulation predisposes to development of thrombotic microangiopathy and that screening patients for such defects could provide informed treatment strategies. PMID- 14566053 TI - Nitric oxide-donating aspirin inhibits beta-catenin/T cell factor (TCF) signaling in SW480 colon cancer cells by disrupting the nuclear beta-catenin-TCF association. AB - Dysregulation of the Wnt pathway and altered Beta-catenin expression are central early events in colorectal carcinogenesis. We studied the ortho, meta, and para (o-, m-, and p-) positional isomers of NO-donating aspirin (NO-ASA), a chemopreventive agent against colon cancer, for their effect on Beta-catenin/T cell factor (TCF) signaling. In human SW480 colon carcinoma cells, cell-growth inhibition by NO-ASA [IC50 values for p-, o-, and m- were 48.1 +/- 4.3 (mean +/ SEM), 60.4 +/- 2.1, and 900 +/-50 microM, respectively] was accompanied by significant inhibition of Beta-catenin signaling. We determined Beta-catenin dependent TCF-4 transcriptional activity by measuring the activity of the luciferase gene placed under the control of TCF-4 regulatory sequences. The IC50 values for Beta-catenin/TCF-4-signaling inhibition by NO-ASA were: o-, 2.6 +/- 0.4; m-, 15 +/- 5; p-, 1.1 +/- 0.1 microM; and for ASA, >5,000 microM. Total or nuclear levels of Beta-catenin and its distribution in the cell were not altered by NO-ASA, as judged by protein expression levels and semiquantitative immunofluorescence analysis. NO-ASA disrupted the association of Beta-catenin and TCF-4 in the nucleus, whereas ASA did not affect it. NO-ASA reduced the expression of cyclin D1, a downstream target gene that plays an important role in colon carcinogenesis. In contrast, a structural analog of NO-ASA lacking the -NO2 moiety did not affect TCF-4 transcriptional activity. Thus, NO-ASA inhibits Beta catenin-mediated TCF activity by preventing the formation of the Beta-catenin/TCF 4 complex. This effect, occurring at NO-ASA concentrations far below those required to inhibit cell growth, may be a critical early event in the chemopreventive activity of NO-ASA against colon cancer. PMID- 14566054 TI - Cardiomyocyte-restricted knockout of STAT3 results in higher sensitivity to inflammation, cardiac fibrosis, and heart failure with advanced age. AB - Cytokines and inflammation have been implicated in the pathogenesis of heart failure. For example, IL-6 family cytokines and the gp130 receptor play important roles in cardiac myocyte survival and hypertrophy. Signal transducer and activator of transcription 3 (STAT3) is a major signaling protein that is activated through gp130. We have created mice with a cardiomyocyte-restricted deletion of STAT3. As measured by serial echocardiograms, mice with cardiac specific deletion of STAT3 are significantly more susceptible to cardiac injury after doxorubicin treatment than age-matched controls. Intriguingly, STAT3 appears to have a critical role in protection of inflammation-induced heart damage. STAT3-deficient mice treated with lipopolysaccharide demonstrated significantly more apoptosis than their WT counterparts. At the cellular level, cardiomyocytes with STAT3 deleted secrete significantly more tumor necrosis factor in response to lipopolysaccharide than those with WT STAT3. Furthermore, histologic examination of the cardiomyocyte-restricted STAT3-deficient mice reveals a dramatic increase in cardiac fibrosis in aged mice. Although no overt signs of heart failure are present in young STAT3-deficient mice, they spontaneously develop heart dysfunction with advancing age. These results indicate the crucial functions of STAT3 in cardiomyocyte resistance to inflammation and other acute injury and in pathogenesis of age-related heart failure. PMID- 14566055 TI - Extracellular signal-regulated kinase 2 is necessary for mesoderm differentiation. AB - The extracellular signal-regulated kinase (ERK) is a component of the mitogen activated protein kinase cascade. Exon 2 of erk2 was deleted by homologous recombination and resulted in embryonic lethality at embryonic day 6.5. erk2 mutant embryos did not form mesoderm and showed increased apoptosis but comparable levels of BrdUrd incorporation, indicating a defect in differentiation. erk2 null embryonic stem (ES) cells exhibited reduced total ERK activity upon serum stimulation, augmented ERK1 phosphorylation, and decreased downstream p90Rsk phosphorylation and activity; yet ES cell proliferation was unaffected. Mutant ES cells were capable of forming mesoderm; however, treatment of mutant ES cells with the mitogen-activated protein kinase kinase inhibitor PD184352 decreased total ERK activity and expression of the mesodermal marker brachyury, suggesting that ERK1 can compensate for ERK2 in vitro. Normal embryos at embryonic day 6.5 expressed activated ERK1/2 in the extraembryonic ectoderm, whereas erk2 mutant embryos had no detectable activated ERK1/2 in this region, suggesting that activated ERK1 was not expressed, and therefore cannot compensate for loss of ERK2 in vivo. These data indicate that ERK2 plays an essential role in mesoderm differentiation during embryonic development. PMID- 14566056 TI - Proteorhodopsin genes are distributed among divergent marine bacterial taxa. AB - Proteorhodopsin (PR) is a retinal-binding bacterial integral membrane protein that functions as a light-driven proton pump. The gene encoding this photoprotein was originally discovered on a large genome fragment derived from an uncultured marine gamma-proteobacterium of the SAR86 group. Subsequently, many variants of the PR gene have been detected in marine plankton, via PCR-based gene surveys. It has not been clear, however, whether these different PR genes are widely distributed among different bacterial groups, or whether they have a restricted taxonomic distribution. We report here comparative analyses of PR-bearing genomic fragments recovered directly from planktonic bacteria inhabiting the California coast, the central Pacific Ocean, and waters offshore the Antarctica Peninsula. Sequence analysis of an Antarctic genome fragment harboring PR (ANT32C12) revealed moderate conservation in gene order and identity, compared with a previously reported PR-containing genome fragment from a Monterey Bay gamma proteobacterium (EBAC31A08). Outside the limited region of synteny shared between these clones, however, no significant DNA or protein identity was evident. Analysis of a third PR-containing genome fragment (HOT2C01) from the North Pacific subtropical gyre showed even more divergence from the gamma proteobacterial PR-flanking region. Subsequent phylogenetic and comparative genomic analyses revealed that the Central North Pacific PR-containing genome fragment (HOT2C01) originated from a planktonic alpha-proteobacterium. These data indicate that PR genes are distributed among a variety of divergent marine bacterial taxa, including both alpha- and gamma-proteobacteria. Our analyses also demonstrate the utility of cultivation-independent comparative genomic approaches for assessing gene content and distribution in naturally occurring microbes. PMID- 14566057 TI - Predicting protein functions from redundancies in large-scale protein interaction networks. AB - Interpreting data from large-scale protein interaction experiments has been a challenging task because of the widespread presence of random false positives. Here, we present a network-based statistical algorithm that overcomes this difficulty and allows us to derive functions of unannotated proteins from large scale interaction data. Our algorithm uses the insight that if two proteins share significantly larger number of common interaction partners than random, they have close functional associations. Analysis of publicly available data from Saccharomyces cerevisiae reveals >2,800 reliable functional associations, 29% of which involve at least one unannotated protein. By further analyzing these associations, we derive tentative functions for 81 unannotated proteins with high certainty. Our method is not overly sensitive to the false positives present in the data. Even after adding 50% randomly generated interactions to the measured data set, we are able to recover almost all (approximately 89%) of the original associations. PMID- 14566060 TI - Probable extirpation of a breeding colony of Short-tailed Albatross (Phoebastria albatrus) on Bermuda by Pleistocene sea-level rise. AB - Albatrosses (Diomedeidae) do not occur in the North Atlantic Ocean today except as vagrants, although five species were present in the early Pliocene. No fossil breeding sites of albatrosses were known previously. The timing of extinction of albatrosses in the North Atlantic was likewise unknown. Deposits that formed near present-day sea level along the southeastern shore of Bermuda contain remains of a former breeding colony and include intact eggshells and bones of embryos, juveniles, and adults of Short-tailed Albatross (Phoebastria albatrus), a critically endangered species now confined to a few islets in the northwestern Pacific Ocean. These deposits are correlated with the middle Pleistocene Lower Town Hill Formation, which at other sites have a radiometric age of 405,000 years ago. This equates with the marine isotope stage 11 interglacial, which culminated in a rise in sea-level to >+20 m. Bones of a juvenile Short-tailed Albatross were also found in beach deposits at +21.3 m from this same interglacial. We interpret the extirpation of albatrosses on Bermuda as probably resulting from lack of nesting sites protected from storm surges over the little emergent land that remained at the height of the marine isotope stage 11 sea level rise. PMID- 14566058 TI - A subset of familial colorectal neoplasia kindreds linked to chromosome 9q22.2 31.2. AB - Colorectal cancer is the second most leading cause of cancer death among adult Americans. Two autosomal dominant hereditary forms of the disease, familial adenomatous polyposis and hereditary nonpolyposis colorectal cancer, together account for perhaps 5% of all cases. However, in approximately 20% of additional colon cancer cases, the affected individuals report a family history of colon cancer in a first-degree relative. Similar familial clusters of colon cancer and early-onset colon adenomas have also been reported. To determine whether such familial aggregations arise by chance or reflect a hereditary colon cancer susceptibility, we conducted a whole genome scan to test for genetic linkage in 53 kindreds in which two or more siblings were affected by age 65 or younger with colon cancer or with advanced colon adenomas that were >1 cm in size or that showed high-grade dysplasia. In this cohort we found genetic linkage of disease (P = 0.00045) to chromosomal region 9q22.2-31.2 in a pattern consistent with autosomal dominant disease alleles. These data suggest that a single locus can contribute to disease susceptibility in a subset of patients with nonsyndromic forms of familial colorectal neoplasia. PMID- 14566059 TI - Immunodominance among herpes simplex virus-specific CD8 T cells expressing a tissue-specific homing receptor. AB - The study of immunodominance within microbe-specific CD8 T cell responses has been challenging. We used a previously undescribed approach to create unbiased panels of CD8 cytotoxic T lymphocyte clones specific for herpes simplex virus type 2, a pathogen with a complex genome encoding at least 85 polypeptides. Circulating herpes simplex virus type 2-specific cells were enriched and cloned after sorting for expression of the skin homing-associated receptor, cutaneous lymphocyte-associated antigen, bypassing restimulation with antigen. The specificity of the resultant cytotoxic clones was determined. Clonal frequencies were compared with each other and with the total number of cytotoxic clones. For each subject within the homing receptor-positive compartment, the CD8 cytotoxic response was dominated by T cells specific for only a few peptides. Previously undescribed antigens and epitopes in viral tegument, capsid, or scaffold proteins were immunodominant in some subjects. Clone enumeration analyses were confirmed in some subjects with dominance studies by using herpes simplex mutants, vaccinia recombinants, and/or enzyme-linked immune spots. We conclude that among circulating cells expressing a homing-associated receptor, during chronic herpes type 2 infection, the CD8 T cell response becomes quite focused despite the presence of many potential antigenic peptides. PMID- 14566061 TI - Exploiting luminescence spectroscopy to elucidate the interaction between sugar and a tryptophan residue in the lactose permease of Escherichia coli. AB - The crystal structure of the Escherichia coli lactose permease at 3.5 A with a bound substrate has been reported recently. The structure reveals the sugar protein contacts, which include hydrophobic stacking between the galactopyranosyl ring of substrate and the indole side chain of Trp-151, as proposed previously. The nature of this interaction is studied here by exploiting the luminescence properties of Trp-151 in a mutant devoid of other tryptophan residues. The following phenomena are observed. (i) The fluorescence emission spectrum of Trp 151 and fluorescence-quenching experiments with water-soluble quenchers demonstrate that Trp-151 is in a hydrophilic environment. (ii) Substrate binding leads to a blue shift in the emission spectrum and reduction in accessibility to polar quenchers, indicating that Trp-151 becomes less exposed to aqueous solvent. (iii) The phosphorescence spectrum of Trp-151 is red-shifted in the presence of substrate, indicating charge separation of the triplet state due to a direct stacking interaction between the galactopyranosyl and indole rings. The spectroscopic data fully complement the x-ray structure and demonstrate the feasibility of fluorescence spectroscopy for studying sugar-protein interactions. PMID- 14566062 TI - The genome of Nanoarchaeum equitans: insights into early archaeal evolution and derived parasitism. AB - The hyperthermophile Nanoarchaeum equitans is an obligate symbiont growing in coculture with the crenarchaeon Ignicoccus. Ribosomal protein and rRNA-based phylogenies place its branching point early in the archaeal lineage, representing the new archaeal kingdom Nanoarchaeota. The N. equitans genome (490,885 base pairs) encodes the machinery for information processing and repair, but lacks genes for lipid, cofactor, amino acid, or nucleotide biosyntheses. It is the smallest microbial genome sequenced to date, and also one of the most compact, with 95% of the DNA predicted to encode proteins or stable RNAs. Its limited biosynthetic and catabolic capacity indicates that N. equitans' symbiotic relationship to Ignicoccus is parasitic, making it the only known archaeal parasite. Unlike the small genomes of bacterial parasites that are undergoing reductive evolution, N. equitans has few pseudogenes or extensive regions of noncoding DNA. This organism represents a basal archaeal lineage and has a highly reduced genome. PMID- 14566064 TI - Evolutionary specialization of a tryptophan indole group for transition-state stabilization by eukaryotic transglutaminases. AB - Covalent posttranslational protein modifications by eukaryotic transglutaminases proceed by a kinetic pathway of acylation and deacylation. Ammonia is released as the acylenzyme is formed, whereas the cross-linked product is released later in the deacylation step. Superposition of the active sites of transglutaminase type 2 (TG2) and the structurally related cysteine protease, papain, indicates that in the formation of tetrahedral intermediates, the backbone nitrogen of the catalytic Cys-277 and the N1 nitrogen of Trp-241 of TG2 could contribute to transition-state stabilization. The importance of this Trp-241 side chain was demonstrated by examining the kinetics of dansylcadaverine incorporation into a model peptide. Although substitution of the Trp-241 side chain with Ala or Gly had only a small effect on the Michaelis constant Km (1.5-fold increase), it caused a >300-fold lowering of the catalytic rate constant kcat. The wild-type and mutant TG2-catalyzed release of ammonia showed kinetics similar to the kinetics for the formation of cross-linked product, indicating that transition state stabilization in the acylation step was rate-limiting. In papain, a Gln residue is at the position of TG2-Trp-241. The conservation of Trp-241 in all eukaryotic transglutaminases and the finding that W241Q-TG2 had a much lower kcat than wild-type enzyme suggest evolutionary specialization in the use of the indole group. This notion is further supported by the observation that transition state-stabilizing side chains of Tyr and His that operate in some serine and metalloproteases only partially substituted for Trp. PMID- 14566063 TI - A presenilin dimer at the core of the gamma-secretase enzyme: insights from parallel analysis of Notch 1 and APP proteolysis. AB - Notch receptors and the amyloid precursor protein are type I membrane proteins that are proteolytically cleaved within their transmembrane domains by a presenilin (PS)-dependent gamma-secretase activity. In both proteins, two peptide bonds are hydrolyzed: one near the inner leaflet and the other in the middle of the transmembrane domain. Under saturating conditions the substrates compete with each other for proteolysis, but not for binding to PS. At least some Alzheimer's disease-causing PS mutations reside in proteins possessing low catalytic activity. We demonstrate (i) that differentially tagged PS molecules coimmunoprecipitate, and (ii) that PS N-terminal fragment dimers exist by using a photoaffinity probe based on a transition state analog gamma-secretase inhibitor. We propose that gamma-secretase contains a PS dimer in its catalytic core, that binding of substrate is at a site separate from the active site, and that substrate is cleaved at the interface of two PS molecules. PMID- 14566065 TI - Effect of some process parameters in enzymatic dyeing of wool. AB - This article reports on the dyeing of wool using an enzymatic system comprising laccase; dye precursor, 2,5-diaminobenzenesulfonic acid; and dye modifiers, catechol and resorcinol. Enzymatic dyeing was performed as a batchwise process at the temperature and pH of maximum enzyme activity. The effects of the process variables reaction time, enzyme, and modifier concentration on fabric color were studied, according to an appropriate experimental design. Different hues and depths of shades could be achieved by varying the concentration of the modifiers and the time of laccase treatment. The duration of the enzymatic reaction appeared to be the most important factor in the dyeing process. Thus, the dyeing process, performed at low temperature and mild pH, was advantageous in terms of reduced enzyme and chemical dosage. PMID- 14566066 TI - Growth and pectinase production by Aspergillus Mexican strain protoplast regenerated under acidic stress. AB - Protoplasts from Aspergillus sp. FP-180 and Aspergillus awamori NRRL- 3112 were released and regenerated at extreme acidic conditions. The best conditions for protoplast release were 0.8 M KCI, pH 5.8, and 3 h of digestion using mycelia from 12- to 16-h cultures from either Aspergillus sp. FP-180 or A. awamori NRRL 3112. The addition of fresh mycelia to an ongoing digestion after 1 h increased protoplast 4.5-5 times. A regeneration efficiency of 90% was attained at pH 6.0, and it was possible to regenerate protoplasts at pH 1.7 with a regeneration efficiency of 0.5% for Aspergillus sp. FP-180. The LpH-10 strain, derived from protoplast from Aspergillus sp FP-180, was able to regenerate at pH 1.7 and grow at pH values as low as 1.5, values at which the original strain is unable to grow. Regeneration at extreme pH improved the performance of LpH-10 strain. It showed a twofold increase in cell growth at pH 2.0 in liquid culture and a higher pectinolytic activity in relation to that produced by the original strain. PMID- 14566067 TI - Production, purification, and biochemical characterization of two beta glucosidases from Sclerotinia sclerotiorum. AB - The filamentous fungus Sclerotinia sclerotiorum produces beta-glucosidases in liquid culture with a variety of carbon sources, including cellulose (filter paper), xylan, barley straw, oat meal, and xylose. Analysis by native polyacrylamide gel electrophoresis (PAGE) followed by an activity staining with the specific chromogenic substrate, 5-bromo 4-chloro 3-indolyl beta-1,4 glucoside (X-glu) showed that two extracellular beta-glucosidases, designated as beta-glu1 and beta-glu2, were in the filter paper culture filtrate. Only one enzyme designated as beta-glu x was revealed by the same method in the xylose culture filtrate. Beta-glu1 and beta-glu2 were purified to homogeneity. The purification procedure consist of a common step of anion-exchange chromatography on DEAE Sepharose CL6B, both high-performance liquid chromatography (HPLC) anion-exchange and gel filtration columns for beta-glu1 and only HPLC gel filtration for beta glu2. Beta-glu1 has a molecular mass of 196 kDa and 96.5 kDa, as estimated by gel filtration and sodium dodecyl sulfate (SDS)-PAGE, respectively, suggesting that the native enzyme may consist of two identical subunits. The same analysis showed that beta-glu2 is a monomeric protein with an apparent molecular mass of about 76.5 kDa. Beta-glu1 and beta-glu2 hydrolyses PNPGlc and cellobiose, with apparent Km values respectively for PNPGlc and cellobiose of 0.1 and 1.9 mM for beta-glu1 and 2.8 and 8 mM for beta-glu2. Both enzymes exhibit the same temperature and pH optima for PNPGlc hydrolysis (60 degrees C and pH 5.0). beta-glu1 was stable over a pH range of 3-8 and kept 50% of its activity after 30 min of heating at 60 degrees C without substrate. It was further characterized by studying the effect of some cations and various reagents on its activity. PMID- 14566068 TI - Bioelectrocatalysts: engineered oxidoreductase system for utilization of fumarate reductase in chemical synthesis, detection, and fuel cells. AB - Fumarate reductase was used as a model oxidoreductase to demonstrate continuous electrical cofactor reduction-oxidation during the bioelectrochemical synthesis and detection of chemicals. The enzyme preparation was immobilized onto a graphite felt electrode that was modified with carboxymethylcellulose (CMC). Nicotinamide adenine dinucleotide (NAD), neutral red, and fumarate reductase (which contained menaquinone) were covalently linked by peptide bonds to the CMC. The electron mediator neutral red allowed NAD and menaquinone to be recycled electrically during enzymatic chemical synthesis. Succinate detection by the bioelectrocatalyst was linear from 5 microM to 10 mM succinate. Fumarate synthesis using this bioelectrode was dependent on succinate utilization and resulted in proportional production of electricity and fumarate. Succinate synthesis using this bioelectrocatalyst was dependent on current and fumarate concentration. This bioelectrocatalyst system may enhance the utility of menaquinone- and/or pyridine nucleotide-linked oxidoreductases in diverse enzymatic fuel cells and sensors. It may also enhance the utility of oxidoreductase-based chemical synthesis systems because it eliminates the problem of cofactor recycling. PMID- 14566069 TI - An integrated process for purification of lysozyme, ovalbumin, and ovomucoid from hen egg white. AB - This article describes an integrated process for simultaneous purification of lysozyme, ovalbumin, and ovomucoid from hen egg white. The crude egg white extract was passed through a cation exchanger Streamline trade mark SP and the bound lysozyme was eluted with 5% ammonium carbonate, pH 9.0, containing 1 M NaCl after elution of avidin. This partially purified lysozyme was further purified 639-fold on dye-linked cellulose beads. Ovalbumin and ovomucoid did not bind to Streamline SP. Ovalbumin could be precipitated from this unbound fraction by 5% trichloroacetic acid, and ovomucoid was removed from the supernatant by precipitation with ethanol. The yields of lysozyme, ovomucoid, and ovalbumin were 77, 94, and 98%, respectively. All the purified proteins showed single bands on sodium dodecyl sulfate polyacrylamide gel electrophoresis. All the steps are easily scalable, and the process described here can be used for large-scale simultaneous purification of these proteins in the pure form. PMID- 14566070 TI - Brain-gut axis in pancreatic secretion and appetite control. AB - The stimulation of exocrine pancreatic secretion that has been attributed by Pavlov exclusively to various reflexes (nervism), was then found that it depend also on numerous enterohormones, especially cholecystokinin (CCK) and secretin, released by duodeno-jejunal mucosa and originally believed to act via an endocrine pathway. Recently, CCK and other enterohormones were found to stimulate the pancreas by excitation of sensory nerves and triggering vago-vagal and entero pancreatic reflexes. Numerous neurotransmitters and neuropeptides released by enteric nervous system (ENS) of gut and pancreas have been also implicated in the regulation of exocrine pancreas. This article was designed to review the contribution of vagal nerves and entero-hormones, especially CCK and other enterohormones, involved in the control of appetitive behavior such as leptin and ghrelin and pancreatic polypeptide family (peptide YY and neuropeptide Y). Basal secretion shows periodic fluctuations with peals controlled by ENS and by motilin and Ach. Plasma ghrelin, that is considered as hunger hormone, increases under basal conditions, while plasma leptin falls to the lowest level. Postprandial pancreatic secretion, classically divided into cephalic, gastric and intestinal phases, involves predominantly CCK, which under physiological conditions acts almost entirely by activation of vago-vagal reflexes to stimulate the exocrine pancreas, being accompanied by the fall in plasma ghrelin and increase of plasma leptin, reflecting feeding behavior. We conclude that the major role in postprandial pancreatic secretion is played by vagus and gastrin in cephalic and gastric phases and by vagus in conjunction with CCK and secretin in intestinal phase. PP, PYY somatostatin, leptin and ghrelin that affect food intake appear to participate in the feedback control of postprandial pancreatic secretion via hypothalamic centers. PMID- 14566071 TI - Nitric oxide synthase activity in blood vessels of spontaneously hypertensive rats: antioxidant protection by gamma-tocotrienol. AB - Involvement of free radicals and nitric oxide (NO) has long been implicated to the pathogenesis of essential hypertension. Several studies using antioxidants as the radical scavenger have shown to confer protection against free radical mediated diseases. This study is designed to investigate the role of antioxidant gamma-tocotrienol on endothelial nitric oxide synthase (NOS) activity in spontaneously hypertensive rats (SHR). SHR's were divided into four groups namely untreated SHR (HC), treatment with 15 mg gamma-tocotrienol/kg diet (gammal), 30 mg gamma-tocotrienol/kg diet (gamma2) and 150 mg gamma-tocotrienol/kg diet (gamma3) and studied for three months. Wister Kyoto (WKY) rats were used as the control (C). Blood pressure was recorded every fortnightly by tail plethysmography. Animals were sacrificed and NOS activity in blood vessels was measured by [3H]arginine radioactive assay. Nitrite concentration in plasma was determined by Greis assay and lipid peroxides in the blood vessels by spectrofluorometry. This study showed that gamma-tocotrienol significantly reduced systolic blood pressure (SBP) in SHRs with a maximum reduction in group treated with gamma-tocotrienol 15 mg/kg diet (HC: 210 +/- 9 mmHg, gammal:123 +/- 19 mmHg). Blood vessels from untreated SHR showed a reduced NOS activity compare to that of WKY rats (C: 1.54 +/- 0.26 pmol/mg protein, HC: 0.87 +/- 0.23 pmol/mg protein; p<0.001). Gamma-tocotrienol improves NOS activity in all the groups with more significance in group gamma2 (p<0.001) and gamma3 (p<0.05). Plasma level of nitrite was reduced in SHR from 55 +/- 3 microM/ml in WKY to 26+/-2 muM/ml (p<0.001). Plasma nitrite level was reversed by treatment with gamma-tocotrienol. (gammal: p<0.001, gamma2: p<0.005, gamma3: p<0.001, respectively). In all the treatment groups, NOS activity showed significant negative correlation with blood pressure (gammal: r=-0.716, p<0.05; gamma2: r=-0.709, p<0.05; gamma3: r=-0.789, p<0.05). For plasma nitrite, although it shows a negative correlation with blood pressure it was significant only in gammal (r=-0.676, p<0.05) and gamma2 (r= 0.721, p<0.05). From this study we found that compared to WKY rats, SHR has lower NOS activity in blood vessels, which upon treatment with antioxidant gamma tocotrienol increased the NO activity and concomitantly reduced the blood pressure. These findings further strengthen the hypothesis that free radicals and NO play critical role in pathogenesis of essential hypertension. PMID- 14566072 TI - Stress-induced rise in endothelaemia, von Willebrand factor and hypothalamic pituitary-adrenocortical axis activation is reduced by pretreatment with pentoxifylline. AB - Stress is considered to be a risk factor of several diseases. The following hypotheses were tested: (1) single exposure to an intensive stressor is followed by endothelial stimulation and/or damage to endothelial cells, (2) potential stress-induced endothelial cell damage is reduced by repeated pretreatment with pentoxifylline and (3) pentoxifylline treatment modifies neuroendocrine activation during stress reflected by changes in hypothalamic-pituitary adrenocortical (HPA) axis function. Rats were treated with saline or pentoxifylline (20 mg/kg, s.c.) once daily for 7 days and then exposed to single immobilization stress for 20 or 120 min. In saline pretreated rats, stress exposure was followed by a rise in endothelaemia, von Willebrand factor concentrations, adrenocorticotropic hormone (ACTH) and corticosterone release, as well as by enhanced gene expression of hypothalamic corticotropin releasing factor (CRH). Stress-induced changes were reduced by pretreatment with pentoxifylline. Significant inhibition was observed in endothelaemia, plasma ACTH and corticosterone concentration in the adrenals. Thus, signs of endothelial injury as well as stress-induced hormone levels were reduced by pretreatment with pentoxifylline, although there is no evidence for a causal relationship. This protective action of pentoxifylline might be of benefit in the prevention and therapy of some stress-related disorders. PMID- 14566073 TI - EPR investigation of in vivo inhibitory effect of guanidine compounds on nitric oxide production in rat tissues. AB - The aim of the present study was to evaluate in vivo effects on NO production of pharmacologically widely used, commercially available NOS inhibitors, structurally related to guanidine. We compared the NO inhibitory potency and selectivity of L-NAME, aminoguanidine and guanabenz in tissues of normal and LPS stimulated rats using ex vivo EPR measurements of the NO radical in its complex with dithiocarbamate-Fe(II). The tissues studied were the brain cortex, kidney, liver, heart and testis. Differential inhibitory effects were seen for L-NAME, aminoguanidine and guanabenz when applied during basal or LPS-stimulated conditions. Aminoguanidine exerted inhibition of NO only after stimulation with LPS. Guanabenz had little effect on NO in liver, kidney, testis and heart under normal conditions, while it reduced the basal NO in brain cortex. After stimulation with LPS guanabenz afforded a partial inhibition of the NO formation in all tissues studied. L-NAME was a potent inhibitor of NO synthesis in all tested tissues, both during basal and LPS stimulated conditions. Our results show that compounds containing a guanidine moiety might possess different NOS inhibitory profiles in vivo. PMID- 14566074 TI - Characterization of "unhealed gastric ulcers" produced with chronic exposure of acetic acid ulcers to indomethacin in rats. AB - We previously discovered that a 4-wk course of indomethacin delivered to rats with acetic acid ulcers resulted in production of "unhealed gastric ulcers" that persisted for up to 12 wks after treatment cessation. The present study examined the mechanism underlying such "unhealed gastric ulcers" with biochemical and histological procedures. "Unhealed gastric ulcers" were induced with a 4-wk indomethacin treatment (1 mg/kg, twice daily) in rats with acetic acid ulcers. Two and 4 wks after treatment cessation, ulcer size was significantly larger in rats receiving indomethacin compared with control animals. Ulcerated tissue prostaglandin E2 levels were significantly lower during indomethacin treatment, but the levels tended to increase after treatment cessation compared with levels measure in the group receiving vehicle. Myeloperoxidase activity levels were significantly higher during indomethacin treatment; such levels persisted after treatment cessation. Histologically, greater degrees of fibrosis and neutrophil accumulation, as well as a lesser degree of angiogenesis were observed in the "unhealed gastric ulcers" compared to ulcers that healed in a normal fashion. It was concluded that severe fibrosis, persistent neutrophil infiltration, and poor angiogenesis in the ulcer base might represent factors involved in the mechanism underlying production of "unhealed gastric ulcers". PMID- 14566075 TI - Angiogenic activity of porcine granulosa cells co-cultured with endothelial cells in a microcarrier-based three-dimensional fibrin gel. AB - To verify the possible role played by pig granulosa cells in the ovarian angiogenic process, we have developed a reliable in vitro system which allows the evaluation of endothelial sprouting and capillary growth in three-dimensional matrices. Granulosa cells collected from porcine follicles of different size were co-cultured with porcine aortic endothelial cells (PAEC) in a microcarrier-based fibrin gel system; after 2 and 5 days of co-culture, we determined the number and length of all endothelial sprouts; moreover, these parameters were quantified only in capillary-like structures, which were defined as continuous multicellular sprouts at least 200 microm long. In granulosa cells- PAEC co-cultures we observed an increase of angiogenic activity as compared to controls (PAEC alone). Granulosa cells from follicles of different size regulate angiogenesis differently: cells from the small follicle group significantly enhanced endothelial sprouting, while those from the large follicle group favoured mainly capillary elongation. Our observations seem therefore to suggest that the development and growth of thecal vascular bed is controlled by paracrine factors of granulosa cell origin that may induce the formation of a primitive capillary plexus during the early phases of antral follicle growth, which will be remodelled in more advanced phases of follicular development. PMID- 14566076 TI - Hypotensive function of the brain angiotensin-(1-7) in Sprague Dawley and renin transgenic rats. AB - Angiotensin-(1-7) (Ang-[1-7]) is present in the brain of normotensive Sprague Dawley (SD) rats, and its hypothalamic content is elevated in TGRmRen2(27) rats (TGR) with renin dependent transgenic hypertension. The purpose of the present study was to determine the role of intrabrain Ang-(1-7) in the regulation of cardiovascular functions in SD and TGR rats under resting conditions and during haemodynamic challenge produced by rapid bleeding. Two groups of experiments were performed on conscious SD and TGR rats that were chronically instrumented with a lateral cerebral ventricle (LCV) cannula and an intraarterial catheter. Blood pressure (MAP) and heart rate period (Hp=distance between two systolic peaks) were continuously monitored: 1) under resting conditions during an LCV infusion of either artificial cerebrospinal fluid (aCSF, 5 microl/hr) or Ang-(1-7) in aCSF (100 pmol/5 microl/hr), and 2) before and after haemorrhage performed during LCV infusion of either aCSF or Ang-(1-7) antagonist (A-779, 4 nmol/5 microl/hr). Cerebroventricular infusion of Ang-(1-7) did not affect baseline MAP in the SD rats but it caused a significant decrease in blood pressure in the TGR rats. In the control experiments, haemorrhage significantly reduced MAP in the SD and TGR rats and heart rate in the TGR rats. Cerebroventricular infusion of Ang-(1-7) antagonist eliminated posthaemorrhagic hypotension in both strains and bradycardia in the TGR rats. The results indicate that intrabrain Ang-(1-7) may contribute to posthaemorrhagic hypotension and bradycardia. Moreover, the manner in which it centrally regulates the cardiovascular functions in the SD and TGR rats may be considerably different. PMID- 14566077 TI - Cardiac and regional haemodynamic effects of endothelin-1 in rats subjected to critical haemorrhagic hypotension. AB - In the present study, we examined cardiac and regional haemodynamic effects of endothelin-1 (ET-1), a potent vasoconstrictive factor, in a rat model of pressure controlled irreversible haemorrhagic shock resulting in the death of all control animals within 30 min. Experiments were carried out in male ethylurethane anaesthetised Wistar rats subjected to hypotension of 20-25 mmHg, which resulted in bradycardia, an extreme decrease in cardiac index (CI) and an increase in total peripheral resistance index (TPRI), with reductions in renal (RBF), hindquarters (HBF) and mesenteric blood flow (MBF). ET-1 (50, 200 pmol/kg) administered intravenously at 5 min of critical hypotension produced increases in mean arterial pressure (MAP) and heart rate (HR), which were significantly higher than those in normotensive animals, and a 100% survival at 2 h after treatment. The effects were accompanied by a rise in CI, a decrease in TPRI, with increases in RBF and HBF and persistently lowered MBF, and an increase in circulating blood volume 20 min after treatment. The cardiovascular effects of ET-1 were inhibited by the ETA receptor antagonist BQ-123 (1 mg/kg), while the ETB receptor antagonist BQ-788 (3 mg/kg) had no effect. In conclusion, ET-1 acting via ETA receptors produces reversal of haemorrhagic hypotension in rats due to the mobilisation of blood from venous reservoirs, with the improvements in cardiac function and the perfusion of peripheral tissues. PMID- 14566078 TI - Secretory dysfunction of vascular endothelium limits the effect of angiotensin converting enzyme inhibitor quinapril on aggregation of erythrocytes in experimental hypertension. AB - Using automatic erythrocyte aggregometer type MA-1 (Myrenne gmbh, Germany), we investigated the hypothesis that therapeutic effectiveness of quinapril- angiotensin converting enzyme inhibitor (ACEI)--in the treatment of hypertension would correlate with improvement of red blood cell (RBC) aggregability. Experiments were performed on commercially available inbred strain of spontaneously hypertensive male rats (SHR) aged 19-21 weeks. Age-matched normotensive Wistar-Kyoto (WKY) rats genetically related to SHR were used as a control. Aggregability of RBC in hypertensive rats was significantly higher than in control WKY animals. Quinapril (100 microg/kg) administered i.p. for 8 days improved RBC aggregability in normotensive rats but surprisingly not in SHR animals. Beneficial effect of quinapril on RBC aggregation observed in normotensive animals did not occur when this drug was injected in combination with aspirin (1 or 50 mg/kg) or with indomethacin (20 mg/kg) or with L-NAME (10 mg/kg). However, much the same damaging effects on RBC aggregability were observed when aspirin, indomethacin or L-NAME were each administered into normotensive animals without quinapril. In contrast with normotensive rats, aggregability of RBC in SHR was not affected either by quinapril or by indomethacin and by L-NAME, given separately or in combination. The only compound significantly worsening RBC aggregability in SHR was aspirin but this effect was not dose-dependent. Quinapril-induced improvement of RBC aggregability in normotensive rats (but not in SHR) was completely abolished by simultaneous administration of B2 receptor antagonist icatibant and successfully mimicked by 8 days of treatment with bradykinin. In vitro aggregability of RBC isolated from WKY was not affected by previous incubation (30 min at 37 degrees C) with quinapril, indomethacin or L-NAME. Only aspirin (3 mM) significantly increased RBC aggregability as compared to placebo. It is concluded that under physiological conditions quinapril efficiently inhibits RBC aggregability and this effect is modulated by secretion of endothelial mediators, mainly prostacyclin and nitric oxide. In hypertension quinapril, in spite of lowering of arterial blood pressure, is unable to display its beneficial effects on RBC aggregability possibly due to the hypertension-induced/accompanied dysfunction of vascular endothelium. Aspirin revealed unique erythrocyte damaging properties, presumably independent of inhibition of cyclooxygenase but related to a direct membrane protein acetylation. PMID- 14566079 TI - Voltage dependent activation of tonic contraction in cardiac myocytes. AB - Contractions of isolated single myocytes of guinea pig heart stimulated by rectangular depolarizing pulses consist of a phasic component and a voltage dependent tonic component. In this study we analyzed the mechanism of activation of the graded, sustained contractions elicited by slow ramp depolarization and their relation to the components of contractions elicited by rectangular depolarizing pulses. Experiments were performed at 37 degrees C in ventricular myocytes of guinea pig heart. Voltage-clamped myocytes were stimulated by the pulses from the holding potential of -40 to +5 mV or by ramp depolarization shifting voltage within this range within 6 s. [Ca2+]i was monitored as fluorescence of Indo 1-AM and contractions were recorded with the TV edge tracking system. Myocytes responded to the ramp depolarization between -25 and -6 mV by the slow, sustained increase in [Ca2+]i and shortening, the maximal amplitude of which was in each cell similar to that of the tonic component of Ca2+ transient and contraction. The contractile responses to ramp depolarization were blocked by 200 microM ryanodine and Ca2+-free solution, but were not blocked by 20 microM nifedipine or 100-200 microM Cd2+ and potentiated by 5 mM Ni2+. The responses to ramp depolarization were with this respect similar to the tonic but not to the phasic component of contraction: both components were blocked by 200 microM ryanodine, and were not blocked by Cd2+ or Ni2+ despite complete inhibition of the phasic Ca2+ current. However, the phasic component but not the tonic component of contraction in cells superfused with Ni2+ was inhibited by nifedipine. Both components of contraction were inhibited by Ca2+-free solution superfused 15 s prior to stimulation. CONCLUSIONS: In myocytes of guinea pig heart the contractile response to ramp depolarization is equivalent to the tonic component of contraction. It is activated by Ca2+ released from the sarcoplasmic reticulum by the ryanodine receptors. Their activation and inactivation is voltage dependent and it does not depend on the Ca2+ influx by the Ca2+ channels or reverse mode Na+/Ca2+ exchange, however, it may depend on Ca2+ influx by some other, not yet defined route. PMID- 14566080 TI - The role of reactive oxygen species and capsaicin-sensitive sensory nerves in the pathomechanisms of gastric ulcers induced by stress. AB - Gastric microcirculation plays an important role in the maintenance of the gastric mucosal barrier and mucosal integrity. Sensory nerves are involved in the regulation of mucosal blood circulation and mucosal defense. Therefore, the ablation of these nerves by neurotoxic doses of capsaicin provides the possibility of determination of their role in gastric mucosal integrity. Stress ulceration represents a serious gastric lesions. Results of our previous experiments have indicated that water immersion and restraint stress (WRS) led to increased oxidative metabolism. Ablation of sensory nerves by high doses of capsaicin retards healing of gastric ulcers, but the role of reactive oxygen species (ROS) in the healing process has been little studied. Therefore, the aim of our present investigations was to determine the participation of ROS in sensory nerve activity during WRS. Experiments were carried out on 90 male Wistar rats and the area of gastric lesions was measured by planimetry. Colorimetric assays were used to determine gastric mucosal levels of malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE), as well as superoxide dismutase (SOD) activity. We demonstrated that inactivation of sensory nerves resulted in magnification of gastric mucosal damage induced by the WRS. In this process, oxidative stress, as reflected by an increase of MDA and 4-HNE tissue concentrations (an index of lipid peroxidation), as well as decrease of SOD activity, could play an important role. Aspirin, applied in a low dose, exerts a protective activity, possibly due to its metabolites, which possess the anti-oxidant and ROS scavanging properties. Pentoxyfilline-induced gastroprotection and hyperemia depends upon attenuation of the oxidative stress. This protection and hyperemia were, at least in part, attenuated by ASA. PMID- 14566081 TI - Poly-C specific ribonuclease activity correlates with increased concentrations of IL-6, IL-8 and sTNFR55/sTNFR75 in plasma of patients with acute pancreatitis. AB - Plasma pancreatic-type Poly-C specific ribonuclease (P-RNase)-enzyme activity increases in patients with acute pancreatitis (AP) who develop pancreatic necrosis and severe disease course. It is considered as a marker of pancreatic tissue destruction. The aim of this study was to estimate interrelations between major inflammatory cytokines such as: interleukin 6 (IL-6), interleukin 8 (IL-8) and tumor necrosis factor soluble receptors: sTNFR55 and sTNFR75 output, and plasma P-RNase activity. The study was carried out in a group of 56 patients with AP, where 20 developed pancreatic necrosis. It was found that serum P-RNase concentration and levels of all studied inflammatory cytokines significantly increase already in the first day from diagnose of the disease (2.5 folds for P RNase, 20 for IL-8, about 200 for IL-6 and 1.5 for receptors, respectively). In the first day from admission to hospital, P-RNase activity significantly correlated with plasma concentration of studied inflammatory cytokines. The most pronounced correlation was found for P-RNase and IL-6 in days 1-4 from diagnose, manifested by Pearson correlation r coefficients amounting to 0.86, 0.79, 0.60 and 0.57 respectively (p<0.001). Dividing the studied AP patients into two groups, varying in severity of disease a significant differences in P-RNase and IL-6, IL-8 and sTNFR55/sTNFR75 were found. In patients with acute necrotizing pancreatitis P-RNase significantly correlate with levels of major inflammatory cytokines. Carried out studies suggest that activity of P-RNase reflects severity of inflammatory reaction, which is dependent on development of pancreatic injury and tissue necrosis in AP. PMID- 14566082 TI - Repeated handling, restraint, or chronic crowding impair the hypothalamic pituitary-adrenocortical response to acute restraint stress. AB - The purpose of the present study was to assess whether, and to what extent prior handling, restraint or social crowding stress during 3-10 days affects the hypothalamic-pituitary-adrenocortical (HPA) response to an acute short-lasting restraint stress. Also the effect of a feedback inhibitory mechanism of corticosterone in the impairment of HPA axis by these stressors was investigated. Male Wistar rats were pretreated with handling 1 min/day for 3-10 days, restraint 2 times daily for 3-7 days and crowding stress for 7 days before exposure to acute restraint stress in metal tubes for 10 min. Some group of rats received exogenous s.c. corticosterone either once 25 mg/kg or 2 times daily 10 mg/kg for 3-10 days before restraint stress. After the last restraint the rats were decapitated and their trunk blood was collected for the measurement of plasma ACTH and serum corticosterone levels. Handling for 3-7 days, restraint for 3-7 days, and crowding for 7 days and a single pretreatment with corticosterone--all significantly and to a similar extent inhibited the restraint stress-induced increase in ACTH and corticosterone secretion. Chronic pretreatment with corticosterone blunted the restraint stress-induced increase in HPA axis activity. These results indicate that repeated short-lasting stress induced by handling, restraint, or crowding potently attenuates the acute restraint stress induced stimulatory action of the HPA axis. They also indicate adaptive action of moderate stress on the HPA axis response to acute stress. The results also suggest that a short-lasting hypersecretion of corticosterone during psychological stress may induce a prolonged feedback inhibition of the HPA axis activity. The attenuation of HPA axis response by prior handling has also obvious methodological implications. PMID- 14566083 TI - Bone marrow-derived vascular cells in response to injury. AB - Intimal hyperplasia is a key lesion for various vascular disorders such as atherosclerosis, postangioplasty restenosis and transplant arteriopathy. It has widely been accepted that intimal smooth muscle cells (SMC) originate from the medial layer in the same artery. However, recent studies suggest that bone marrow can also provide circulating progenitors for vascular SMC. Bone marrow-derived SMC participate in neointimal formation in animal models of allotransplantation, severe mechanical injury and hyperlipidemia-induced atherosclerosis. In human, transplantation arteriopathy also seems to involve circulating SMC, but their role in atherosclerosis and restenosis remains to be elucidated. Mobilization, differentiation and proliferation steps of SMC progenitors will provide promising targets for novel therapeutic approaches against proliferative vascular diseases. PMID- 14566084 TI - Role of protease-activated receptors in the vascular system. AB - Thrombin is one of the key molecules involved in the development of vascular diseases. Thrombin does not only serve as a coagulation factor, but it also exerts cellular effects by activating protease (proteinase)-activated receptors (PARs), a family of seven-transmembrane G protein-coupled receptors. This study focused on the role of PARs in the vascular system. Among the four members so far identified, PAR-1 and PAR-2 were found to play an important role in the vascular system, while the functional roles of PAR-3 and PAR-4 appear to be mostly limited to platelets. The endothelial cells play a primary role in mediating the vascular effects of PARs under physiological conditions, while PARs of the smooth muscle cells can be induced under pathological conditions, and therefore play a more pathophysiological role. PAR-1 and PAR-2 mediate various vascular effects including regulation of vascular tone, proliferation and hypertrophy of smooth muscle and angiogenesis. Since proteases are activated under pathological conditions such as hemorrhage, tissue damage, and inflammation, PARs are suggested to play a critical role in the development of functional and structural abnormality in the vascular lesion. Understanding the functional role of PARs in the vascular system can thus help in the development of new strategies for the prevention and therapy of vascular diseases. PMID- 14566085 TI - Mechanism of an inhibitory effect of nipradilol on rat vascular smooth muscle cell growth. AB - The aim of this study was to clarify the mechanism of an inhibitory effect of nipradilol on cultured rat vascular smooth muscle cell (VSMC) growth. After being starved, cultured VSMCs were stimulated by 5% fetal bovine serum with various concentrations of nipradilol. Nipradilol dose-dependently decreased the values of [(3)H]-thymidine incorporation, cell numbers and total cellular protein content, and the levels of phosphorylated extracellular signal-regulated protein kinase 1/2 and p38. It also suppressed the level of proliferative cell nuclear antigen in a dose-dependent manner. In contrast, nipradilol did not change the level of the phosphorylated value of c-jun NH(2)-terminal protein kinase or cytoplasmic histone-associated DNA fragments in VSMCs. These results indicate that nipradilol suppresses cell growth without apoptosis in rat VSMCs, suggesting that it could be effective for preventing the progression of restenosis after angioplasty. PMID- 14566086 TI - Decreased plasma adiponectin levels in young obese males. AB - Plasma adiponectin levels are reduced in middle-aged obesity and in patients with type 2 diabetes and coronary artery disease. The purpose of this study was to investigate the effects of early-aged obesity on plasma adiponectin level. Twenty six male college students (19.2 +/- 1.1 years, obese group: n = 15, [body mass index > 25, percent body fat > 25%], non-obese group: n = 11) participated in the present study. We measured anthropometric parameters and plasma adiponectin and leptin level. Plasma adiponectin levels in the obese group were significantly lower than those in the non-obese group (obese: 4.7 +/- 2.0 micro g/ml, non obese: 6.8 +/- 2.2 micro g/ml, p < 0.05). On the other hand, plasma leptin levels in the obese group were significantly higher than those in the non-obese group (obese: 8.4 +/- 3.2 ng/ml, non-obese: 2.6 +/- 2.1 ng/ml, p < 0.001). Plasma adiponectin levels were significantly correlated with body weight (r = -0.415, p < 0.05) and percent body fat (r = -0.412, p < 0.05). Stepwise multiple regression analysis revealed that percent body fat was a significant independent predictor of plasma adiponectin level (r = 0.406, p < 0.05). These results show that obesity is associated with reduced plasma adiponectin even in young subjects. PMID- 14566087 TI - Overexpression of p21Waf1 induces apoptosis in immortalized human vascular smooth muscle cells. AB - To understand the role of the cell cycle regulatory protein in the control of smooth muscle cell (SMC) proliferation, we tested the overexpression of p21Waf1, a cyclin-dependent kinase inhibitor, in human normal (MS9) and immortalized SMCs (ISS10) transfected with ori-minus simian virus 40 DNA, using an adenovirus mediated system. In MS9, overexpression of p21Waf1 resulted in the inhibition of cell cycle progression at the G1/S boundary without apoptosis. On the other hand, in ISS10, overexpression of p21Waf1 induced marked apoptosis. In these cells, immunohistochemistry revealed that overexpressed p21Waf1 was localized in the nucleus. No differential expression pattern of either p53 or SV40T was observed in p21Waf1- and control gene (beta-galactosidase)-infected cells. Old-passaged ISS10 cells eventually showed growth arrest and a senescent-like phenotype. Immunohistochemistry revealed that p21Waf1 was localized in the cytoplasm of the early-passaged cells, but was found in the nucleus of the old-passaged cells. Our data suggested that nuclear accumulation of p21Waf1 plays a role in the cell death of immortalized SMC, which carries dysfunction of the cell cycle regulatory proteins such as p53. This culture model may be useful for studying the process of SMC proliferation, cell death, senescence, and cell cycle regulation. PMID- 14566088 TI - Arterial wall thickness is associated with insulin resistance in type 2 diabetic patients. AB - The aim of the present study was to investigate the independent association of the intimal-medial thickness of carotid and femoral arteries (CA-IMT and FA-IMT), a marker of atheroscelosis, with insulin resistance in type 2 diabetic patients. We evaluated CA-IMT and FA-IMT by high-resolution ultrasonography and insulin resistance determined by euglycemic hyperinsulinemic clamp in 119 type 2 diabetic subjects, 71 males and 48 females (age, 54 +/- 12 (SD) years). In simple regression analyses, CA-IMT and FA-IMT were significantly inversely correlated with insulin sensitivity index (CA-IMT, r = -0.225, p = 0.010; FA-IMT, r = 0.186, p = 0.043, respectively). Multiple regression analysis was performed with the logarithm of CA-IMT or FA-IMT as a dependent variable and insulin sensitivity index as an independent variable along with known clinical risk factors. Insulin sensitivity index exhibited a significant independent contribution to log (CA IMT) (beta = -0.204, p = 0.033) and to log (FA-IMT) (beta = -0.237, p = 0.010) in these models (CA-IMT, R(2) = 0.347, p < 0.0001; FA-IMT, R(2) = 0.398, p < 0.0001, respectively). In conclusion, insulin resistance is associated with both CA-IMT and FA-IMT in type 2 diabetic patients, suggesting that it is an independent risk factor for the development of atherosclerosis in type 2 diabetes. PMID- 14566089 TI - Pulse wave velocity in lower-limb arteries among diabetic patients with peripheral arterial disease. AB - OBJECTIVE: Patients with type 2 diabetes mellitus are at an increased risk of atherosclerosis including peripheral arterial disease (PAD). The purpose of this study was to examine the possible alteration in pulse wave velocity (PWV) in lower-limb arteries among diabetic patients with PAD. METHODS: We measured brachial-ankle PWV (baPWV) using an automatic device in 101 healthy control subjects and 102 type 2 diabetic patients including those with PAD. RESULTS: Diabetic patients without PAD showed a higher baPWV than the healthy control subjects. There was no significant difference in baPWV between the right and left legs in these groups. In contrast, among diabetic patients with PAD, baPWV was significantly lower in the affected legs than in the non-affected legs, and the reduction in baPWV was greater in those with lower ankle-brachial pressure index (ABI). In the patients with PAD who received percutaneous transluminal angioplasty, both baPWV and ABI were increased following successful vessel dilatation. CONCLUSIONS: These results suggest that baPWV is increased in diabetic patients, whereas it is decreased in the affected legs in diabetic patients with PAD. Widening of the right-left difference in baPWV may be a novel marker of PAD. PMID- 14566090 TI - Association of heart rate with coronary risk factors and increased white blood cell counts in healthy Japanese people. AB - Elevated resting heart rate (HR) is associated with hypertension in addition to or similar to increased incidence of cardiovascular morbidity and mortality. Subjects with high HR exhibit the characteristic features of insulin resistance syndrome. Sympathetic overactivity underlies this clinical condition. However, the true mechanism of high HR is unclear. We examined the relation between resting HR and age, gender, traditional coronary risk factors, and white blood cell (WBC) count in urban Japanese. The study population comprised 2,736 people aged 40, 50, and 60 years old. Multiple regression analysis of the total population showed that gender (p < 0.001), systolic and diastolic blood pressure (SBP, DBP; p < 0.001), serum triglycerides (TG; p < 0.001), fasting plasma glucose (FPG; p < 0.001) and WBC count (p < 0.001) had significant correlations with HR in the total population. Exercise was negatively correlated with HR (p < 0.001). In the age-adjusted profiles, gender (p < 0.001), SBP (p < 0.001), FPG (p < 0.001, p < 0.01 at age 40 years) and WBC count (p < 0.001) showed a significant correlation with HR among all three age groups. A negative association between HDL cholesterol (HDLc) and HR was seen at age 60 years (p < 0.01). A negative correlation between exercise and HR was seen at age 40 years (p < 0.001) and age 50 years (p < 0.01). It is suggested that the WBC count is one of the important determinants of resting HR regardless of age. Further studies are necessary to clarify the link between HR, coronary risk factors, and chronic infections. PMID- 14566091 TI - Quantitative assessment of angiogenesis in murine antigen-induced arthritis by intravital fluorescence microscopy. AB - Inhibition of angiogenesis might be a therapeutic approach to prevent joint destruction caused by the overgrowing synovial tissue during chronic joint inflammation. The aim of this study was to investigate angiogenesis in the knee joint of mice with antigen-induced arthritis (AIA) by means of intravital microscopy. In 14 mice (C57BL6/129Sv) intravital microscopic assessment was performed on day 8 after AIA induction in two groups (controls, AIA). Synovial tissue was investigated by intravital fluorescence microscopy using FITC-dextran (150 kD). Quantitative assessment of vessel density was performed according to the following categories: functional capillary density (FCD, vessels <10 microm in diameter), functional vessel density (FVD, vessels >10 microm) and FVD of vessels with angiogenic criteria (convoluted vessels, abrupt changes of diameter, vessels which are generated by sprouting and progressively pruned and remodelled). Microvessel count was performed using immunohistochemistry. There was no significant difference in FCD between the control group (337 +/- 9 cm/cm2; mean +/- SEM) and the AIA group (359 +/- 13 cm/cm2). The density of vessels larger than 10 microm diameter was significantly increased in animals with AIA (135 +/- 10 vs. 61 +/- 5 cm/cm2 in control). The density of blood vessels with angiogenic criteria was enhanced in arthritic animals (79 +/- 17 vs. 12 +/- 2 cm/cm2 in control). There was a significant increase in the microvessel count in arthritic animals (297 +/- 25 vs. 133 +/- 16 mm(-2) in control). These findings demonstrate that angiogenesis in murine AIA can be assessed quantitatively using intravital microscopy. Further studies will address antiangiogenic strategies in AIA. PMID- 14566093 TI - Statistical properties of affected sib-pair linkage tests. AB - Genetic linkage analysis is a powerful tool for the identification of disease susceptibility loci. Among the most commonly applied genetic linkage strategies are affected sib-pair tests, but the statistical properties of these tests have not been well characterized. Here, we present a study of the distribution of affected sib-pair tests comparing the type I error rate and the power of the mean test and the proportion test, which are the most commonly used, along with a novel exact test. In contrast to existing literature, our findings showed that the mean and proportion tests have inflated type I error rates, especially when used with small samples. We developed and applied corrections to the tests which provide an excellent adjustment to the type I error rate for both small and large samples. We also developed a novel approach to identify the areas of higher power for the mean test versus the proportion test, providing a wider and simpler comparison with fewer assumptions about parameter values than existing approaches require. PMID- 14566092 TI - CD31 regulates direction and rate of neutrophil migration over and under endothelial cells. AB - Mechanisms guiding migration of neutrophils through endothelium are poorly understood. We showed previously that CD31-CD31 binding acted as an 'accelerator' for neutrophils migrating on platelets, while neutrophil alpha(v)beta3-integrin acted as a sensor to align migration with the direction of imposed flow. Here, we perfused neutrophils over human umbilical vein endothelial cells (HUVEC) treated with tumour necrosis factor-alpha, and characterised the kinetics of migration over, through and underneath the HUVEC. Before penetrating the monolayer, activated neutrophils migrated relatively slowly over the surface (approximately 6 microm/min), preferentially in the direction of flow. Once transmigrated, neutrophils moved more rapidly (approximately 14 microm/min) without preferred direction. Treatment of HUVEC and/or neutrophils with function-blocking antibodies against CD31 reduced directionality but not velocity of migration on top of HUVEC, and reduced velocity of migration underneath the monolayer. If neutrophils were pre-activated with formyl peptide, they did not migrate through the HUVEC, but migrated with increased velocity and directionality on top. Under these circumstances, both velocity and directionality were reduced by blocking CD31. alpha(v)beta3-integrin did not regulate migration under any conditions. We conclude that CD31-CD31 bonds act as robust sensors which can guide neutrophil migration, and also modify its velocity. Thus mechanical and adhesive signals can regulate neutrophil migration driven by locally-acting chemotactic agents. PMID- 14566094 TI - Polymorphisms of the HDL receptor gene associated with HDL cholesterol levels in diabetic kindred from three populations. AB - OBJECTIVE: We examined polymorphisms in the HDL receptor, SR-BI, for association with plasma HDL cholesterol levels. METHODS: Study subjects, including 847 women and 725 men, were from families originally ascertained for type 2 diabetes from Finland, Sweden and Israel. Four common polymorphisms were examined in linear regression analysis: an exon 1 missense (EX1), exon 8 silent (EX8), intron 5 (IVS5) and intron 10 (IVS10) variants. RESULTS: Genotype combinations for the three polymorphisms in linkage disequilibrium (IVS5, EX8 and IVS10) were found to be associated with HDL-C among women from the Israeli (p = 0.01) and Swedish (p = 0.06) populations. In Finnish women, the association was only apparent after taking into account effect modification by triglyceride levels (p = 0.04). One specific pattern of genotypes, denoted by presence of the IVS5_T and EX8_C alleles, and absence of the IVS10_G allele, was consistently associated with the lowest mean levels of HDL-C in women from all three populations. These same associations were not found in men. CONCLUSIONS: Polymorphic variation of the SR BI gene may influence HDL-C levels and act in a sex-dependent manner. PMID- 14566096 TI - Modeling and E-M estimation of haplotype-specific relative risks from genotype data for a case-control study of unrelated individuals. AB - The US National Cancer Institute has recently sponsored the formation of a Cohort Consortium (http://2002.cancer.gov/scpgenes.htm) to facilitate the pooling of data on very large numbers of people, concerning the effects of genes and environment on cancer incidence. One likely goal of these efforts will be generate a large population-based case-control series for which a number of candidate genes will be investigated using SNP haplotype as well as genotype analysis. The goal of this paper is to outline the issues involved in choosing a method of estimating haplotype-specific risk estimates for such data that is technically appropriate and yet attractive to epidemiologists who are already comfortable with odds ratios and logistic regression. Our interest is to develop and evaluate extensions of methods, based on haplotype imputation, that have been recently described (Schaid et al., Am J Hum Genet, 2002, and Zaykin et al., Hum Hered, 2002) as providing score tests of the null hypothesis of no effect of SNP haplotypes upon risk, which may be used for more complex tasks, such as providing confidence intervals, and tests of equivalence of haplotype-specific risks in two or more separate populations. In order to do so we (1) develop a cohort approach towards odds ratio analysis by expanding the E-M algorithm to provide maximum likelihood estimates of haplotype-specific odds ratios as well as genotype frequencies; (2) show how to correct the cohort approach, to give essentially unbiased estimates for population-based or nested case-control studies by incorporating the probability of selection as a case or control into the likelihood, based on a simplified model of case and control selection, and (3) finally, in an example data set (CYP17 and breast cancer, from the Multiethnic Cohort Study) we compare likelihood-based confidence interval estimates from the two methods with each other, and with the use of the single-imputation approach of Zaykin et al. applied under both null and alternative hypotheses. We conclude that so long as haplotypes are well predicted by SNP genotypes (we use the Rh2 criteria of Stram et al. [1]) the differences between the three methods are very small and in particular that the single imputation method may be expected to work extremely well. PMID- 14566095 TI - Population frequencies of single nucleotide polymorphisms (SNPs) in immuno modulatory genes. AB - Inherited polymorphisms in immuno-modulatory genes may contribute to variations in immune function and genetic susceptibility for complex diseases, including cancer. We report results from a comprehensive study to discover novel single nucleotide polymorphisms (SNPs) and to estimate allelic frequency for both novel and known coding and regulatory region SNPs in genes encoding proteins that have been implicated in the immune response to tumors. We identified 12 novel nucleotide substitution variants and one deletion variant in 17 genes analyzed (TGFBETA;R, BETA;2M, IFNGAMMA;, TNFALPHA;, TNFALPHA;R, LTALPHA;, IL-6, IL-12, IL 2, IL-1ALPHA;, IL-1BETA;, IL-1RN, IL-10, CTLA4, CD40L, FAS and FASL). We determined the frequency of these novel polymorphisms, as well as 17 previously identified polymorphisms, in a control sample of 158 individuals, approximately half of which were Caucasian (n = 74) and half of which were African American (n = 84). Significant differences in allele frequencies were observed between the two racial groups for 13/17 genes tested. These allelic variations maybe associated with alterations in immune function and thus susceptibility to a number of complex disease states such as cancer. PMID- 14566097 TI - A genome-wide scan for loci affecting normal adult height in the Framingham Heart Study. AB - OBJECTIVE: To map loci influencing normal adult height in 335 families from the Framingham Heart Study. METHODS: We analyzed data consisting of 1,702 genotyped individuals who have been followed over time. The first height measurement for individuals between the ages 20-55 years was analyzed in a genome-wide scan using variance component linkage analysis. Sex, age, and cohort effects were removed before analysis. RESULTS: Two regions (18pter-p11, 22q11.2) with multipoint LOD scores >1.0 (-log p values >2.0) were detected: we obtained LOD scores of 1.38 at D18S1364, and of 1.10 at D22S345. Analysis of height as a sex-limited phenotype revealed a peak in the 9p21 region near D9S319 with a maximum LOD score of 1.65 ( log p value >3.0) when only male height phenotypes were used. When only female phenotypes were used, a peak with a maximum LOD score of 1.85 (-log p value of 2.70) was observed in the 11q25-qter region near D11S2359. CONCLUSIONS: Our region of interest on chromosome 9 has been implicated by two prior studies. Variance components analysis appeared to be sensitive to pedigree structures as well as the method of IBD computation used. PMID- 14566098 TI - How to model a complex trait. 1. General considerations and suggestions. AB - Usually, when complex traits are at issue, not only are the loci of the responsible genes a priori unknown; the same also holds for the mode of inheritance of the trait, and sometimes even for the phenotype definition. The term mode of inheritance relates to both the genetic mechanism, i.e., the number of loci implicated in the etiology of the disease, and the genotype-phenotype relation, which describes the influence of these loci on the trait. Having an idea of the genetic model can crucially facilitate the mapping process. This holds especially in the context of linkage analysis, where an appropriate parametric model or a suitable nonparametric allele sharing statistic may accordingly be selected. Here, we review the difficulties with parametric and nonparametric linkage analysis when applied to multifactorial diseases. We address the question why it is necessary to adequately model a genetically complex trait in a linkage study, and elucidate the steps to do so. Furthermore, we discuss the value of including unaffected individuals into the analysis, as well as of looking at larger pedigrees, both with parametric and nonparametric methods. Our considerations and suggestions aim at guiding researchers to genotyping individuals at a trait locus as accurately as possible. PMID- 14566099 TI - Undifferentiated carcinoma with osteoclast-like multinucleated giant cells arising in an ovarian mature cystic teratoma. AB - Malignant transformation is an unusual complication of mature cystic teratomas. Squamous cell carcinomas and adenocarcinomas are predominant among these malignant tumors. We present a 70-year-old Japanese woman with a giant ovarian tumor (30 x 23 x 9 cm) and multiple peritoneal dissemination. The huge tumor was filled with necrotic and fatty tissue and coagula accompanied with hair balls histologically diagnosed as undifferentiated carcinoma with multinucleated giant cells arising in an ovarian mature cystic teratoma. The residual disseminated tumors grew very rapidly and occupied the whole abdominal cavity again 3 weeks after the surgery. The patient died 5 weeks after the operation. PMID- 14566100 TI - Psychosis and antipsychotic medications in Alzheimer's disease: clinical management and research perspectives. AB - Psychosis is common in patients with Alzheimer's disease (AD) and contributes substantially to patient morbidity and caregiver distress. Antipsychotic medications are used to treat psychosis and other psychiatric or behavioral symptoms in AD, although optimal treatment guidelines have been elusive. Choosing the most advantageous medication for an individual patient is challenging. This article provides an overview of clinical management principles and medication treatment strategies for patients with AD and psychosis. Effects of individual medications are also described. Medications in the conventional neuroleptic, atypical antipsychotic, cholinesterase inhibitor, and serotonergic classes have been shown to ameliorate psychosis and behavioral symptoms in patients with AD, although the evidence is not conclusive for many medications. Side effects vary substantially across medication classes and modestly among individual patients. Improvement in agitation, aggression, or other behaviors with antipsychotic medication treatment may not depend on distinct antipsychotic effects. In contrast, there is preliminary evidence that delusions and hallucinations may respond to treatment with medications outside the antipsychotic class. Many important clinical questions warrant further research study. In particular, studies to compare how individual symptoms respond to different medications, and to examine how to best manage overlapping symptoms or incomplete treatment response are needed. PMID- 14566101 TI - Cisplatin ototoxicity involves organ of Corti, stria vascularis and spiral ganglion: modulation by alphaMSH and ORG 2766. AB - It has been shown that alphaMSH and the nonmelanotropic ACTH/MSH(4-9) analog ORG 2766 can ameliorate cisplatin-induced neurotoxicity and ototoxicity. Here, we investigated whether these peptides delay the occurrence of the cisplatin-induced shift in auditory threshold, and whether they affect the subsequent recovery of cochlear potentials. Chronically implanted round window electrodes were used to obtain daily recordings of auditory nerve compound action potentials (CAP) and cochlear microphonics at frequencies ranging from 2 to 16 kHz. Cisplatin (1.5 mg/kg i.p.) plus alphaMSH, ORG 2766 (75 mug/kg s.c.), or saline were injected daily until the 40-dB CAP threshold shift at 8 kHz was reached. Endocochlear potential (EP) was measured either 1-2 days or 28 days later, followed by morphometric analysis of the cochlea. Peptide cotreatment did not consistently delay the threshold shift; however, the CAP threshold recovered faster and to a greater extent, with the potency order being alphaMSH > ORG 2766 > saline. Significant recovery at the 2 highest frequencies was seen in the alphaMSH treated animals only. CAP amplitude at high sound pressures, which depends more on nerve function than on outer hair cell (OHC) function, decreased severely in all groups but recovered significantly in the alphaMSH- and completely in the ORG 2766-cotreated group. EP was significantly lower in the first days after the threshold shift but had completely recovered at 28 days. Morphometric analysis of the spiral ganglion also indicated involvement of ganglion cells. OHC loss was most severe in the basal turn of saline-cotreated animals. These data suggest that the cisplatin-induced acute threshold shift might be due to reversible strial failure, whereas subsequent OHC survival determines the final degree of functional recovery. Both OHC loss and neuronal function were ameliorated by peptide cotreatment. PMID- 14566102 TI - Cortisol levels in the human perilymph after intravenous administration of prednisolone. AB - Cortisone is used in various inner ear disorders such as sudden hearing loss. However, it is not known if the doses of prednisolone employed in therapy increase the cortisol level in the inner ear. To evaluate the level of cortisol within the perilymph after intravenous administration of 125 and 250 mg of prednisolone, serum and perilymph samples of 29 consecutive patients with clinical otosclerosis subjected to stapedectomy were collected. Cortisol levels were determined by RIA. The perilymphatic cortisol level was significantly increased in the group with 250 mg of prednisolone while the perilymphatic cortisol levels were not significantly different between the control group and the patients treated with 125 mg. Although the therapeutic dose of prednisolone is not known, we conclude that the application of 250 mg has a greater impact on the inner ear than 125 mg. PMID- 14566103 TI - Sensitivity of the neuromagnetic N100m deflection to spectral bandwidth: a function of the auditory periphery? AB - The amplitude of the auditory evoked field (AEF) component N100m in response to tonal stimuli of varying spectral bandwidth and periodicity was compared with simulated peripheral activity patterns of the auditory nerve (AN). The AEF of ten subjects was recorded with a 37-channel axial gradiometer system (four independent measurement sessions per hemisphere). The simulated peripheral activity was characterized using measures derived from spike probabilities of the AN. Stimuli were pure tones, narrow-band harmonic complex tones (spectrum: 4-4.8 kHz), and broad-band harmonic complex tones (spectrum: 800 Hz-4.8 kHz) with periodicities of 100, 200, and 400 Hz. The intensity of all stimuli was set to 65 dB above the absolute thresholds. Both the simulated AN activity and measured cortical response amplitudes increased consistently with spectral bandwidth. This suggests that the enhanced sensitivity of the N100m amplitude to broad-band complex tones is to some extent a function of the auditory periphery. PMID- 14566104 TI - Preattentive memory-based comparison of sound intensity. AB - Changes in the intensity of repeated, ignored sounds elicit the mismatch negativity (MMN) brain response which reflects preattentive detection of the change. It is generally assumed that the MMN in response to intensity changes reflects a memory-based comparison mechanism rather than being due to differential states of refractoriness of intensity-specific cortical neurons. In the present study, an experimental protocol consisting of 4 oddball blocks and 1 control block was used in order to separate memory-comparison-related effects from refractoriness-related ones. This design allowed an assessment of intensity MMN using physically identical stimuli with equal probability of occurrence in separate blocks, while avoiding contamination by refractoriness. Results were consistent with an MMN in response to intensity change that reflects genuine memory-based comparison. PMID- 14566105 TI - Intelligibility of sentences recorded from the uterus of a pregnant ewe and from the fetal inner ear. AB - The intelligibility of sentences recorded from the uterus of a pregnant ewe and from the near-term fetal sheep inner ear was judged by 30 listeners. Sentences were presented to the ewe at 95 and 105 dB SPL while sequential recordings of sound with a hydrophone and a cochlear microphonic (CM) with electrodes were made. Recordings were randomized and presented to listeners to judge the intelligibility of sentences processed through the ewe and fetal inner ear. Intelligibility scores were nearly 99% for air and uterus conditions, dropped to 73% for CM ex utero and to 41% for CM in utero. Results indicated that filtering provided by the tissues and fluids of the maternal abdomen did not affect sentence intelligibility significantly, but the filtering effects as the signal passed into the fetal inner ear resulted in much poorer intelligibility. PMID- 14566106 TI - Vascular access surveillance: evaluation of combining dynamic venous pressure and vascular access blood flow measurements. AB - BACKGROUND/AIMS: Vascular access thrombosis is one of the most morbid problems encountered by hemodialysis patients. Surveillance protocols utilizing venous pressure (Vp) and vascular access blood flow (VABF) measurements have been employed to preserve vascular access. We undertook a study to evaluate combined dynamic Vp and VABF measurements in the identification of vascular access impairment. We also assessed the effect of preventive repair on thrombosis rates in impaired vascular accesses identified by surveillance. METHODS: Eighty-six chronic hemodialysis patients with a functioning vascular access were enrolled into the surveillance protocol. All vascular accesses with greater than 50% of monthly Vp readings >120 mm Hg or VABF <500 ml/min in arteriovenous fistulas (AVFs) and VABF <650 ml/min in arteriovenous grafts (AVGs), or a decrease in VABF >25% compared to the highest previously measured value, were considered positive. Stenosis >50% on fistulography or a thrombotic event were defined as a 'vascular access impairment episode' while a stenosis <50% or the absence of a thrombotic event was defined as 'no vascular access impairment episode'. Thrombosis rates and intervention rates were calculated per access year at risk. RESULTS: The sensitivity and specificity of the combined surveillance protocol for AVFs were 73.3 and 91%, respectively. In AVGs, they were 68.8 and 87.5%, respectively. The rate of thrombotic events was lower in patients who underwent early repair. The addition of dynamic Vp did not reduce the thrombosis rate any further than surveillance based on VABF alone. CONCLUSION: Combined monitoring for surveillance of AVFs improved sensitivity but had little benefit in AVGs over VABF monitoring alone. Raising VABF cutoff levels might increase and improve identification of vascular access risk for thrombosis, but at the expense of lower specificity. PMID- 14566107 TI - Are CD34+ cells found in renal interstitial fibrosis? AB - BACKGROUND: The CD34 antigen is a marker of precursor myeloid cells and endothelial cells, but also of a subset of spindle cells. METHODS: To establish the possible role of CD34+ cells in the progression of glomerulonephritis, 30 renal biopsies were investigated. Immunohistochemistry for CD34 and smooth muscle actin (SMA) were performed by standard methods. The results were assessed semiquantitatively. The relative interstitial volume (RIV) was evaluated by the point counting method. Clinical data such as sex, age, serum creatinine and urea levels were taken into account. RESULTS: CD34+ spindle cells and SMA+ myofibroblasts were present in the renal interstitium in all but one case. In membranous, membranoproliferative and segmental-proliferative glomerulonephritis, CD34+ cells were seen in clusters, while in IgA nephropathy and in segmental and focal glomerulosclerosis, single, scattered cells were seen. The density of CD34+ cells showed a positive correlation with the myofibroblast density and the RIV. Serum creatinine and urea levels were associated with the RIV of the kidney and to a smaller degree with the number of myofibroblasts and CD34+ cells. CONCLUSION: These results show that in glomerulonephritis, CD34+ spindle cells appear in the interstitium. At present, their possible role can only be speculated. PMID- 14566108 TI - The journey of a lifetime-from general surgeon to surgeon general. PMID- 14566109 TI - The worst injury predicts mortality outcome the best: rethinking the role of multiple injuries in trauma outcome scoring. AB - BACKGROUND: The prediction of outcome after injury must incorporate measures of injury severity, but there is no consensus on how many injuries should be used in calculating these measures. Initially, the single worst injury was used to predict outcome, but the introduction of the Injury Severity Score allowed up to three injuries to contribute to outcome prediction. Subsequently, other outcome prediction approaches used many (New Injury Severity Score [NISS]) or all (ICISS and Trauma Registry Abbreviated Injury Scale Score [TRAIS], which use International Classification of Diseases, Ninth Revision [ICD-9] and Abbreviated Injury Scale [AIS] survival risk ratios [SRRs], respectively) of a patient's injuries. The ability of only the most severe injury in predicting mortality has never been studied. Our objective was to determine the ability of a patient's worst injury to predict mortality. METHODS: A 10-fold cross-validation design was used to compute six scores for each of 160,208 patients from a large trauma database (the National Trauma Data Bank [NTDB]). The scores were ICISS, TRAIS, ICISS1 (only a patient's worst ICD-9 SRR), TRAIS1 (only a patient's worst AIS SRR), NISS (sum of squares of worst three AIS severity measures), and MAXAIS (worst AIS severity measure). Discrimination was assessed using the area under the receiver operating characteristic curve. Logistic regression R2 gauged the proportion of variance each score explained. The Akaike information criterion, a deviance statistic (lower is better), assessed model fit. RESULTS: The receiver operating characteristic curve, R2, and Akaike information criterion statistics (NC_ICISS and NC_ICDSRR1 represents scores derived from the original North Carolina Hospital Discharge Database SRRs) are summarized in tabular form in the Results section. CONCLUSION: Regardless of scoring type (ICD/AIS SRRs or AIS severity), a patient's worst injury discriminates survival better, fits better, and explains more variance than currently used multiple injury scores. PMID- 14566110 TI - Multiple organ failure in trauma patients. AB - SUMMARY: BACKGROUND As care of the critically ill patient has improved and definitions of organ failure have changed, it has been observed that the incidence of organ failure and the mortality associated with organ failure appear to be decreasing. In addition, many early studies included large heterogeneous populations of both medical and surgical patients that may have influenced the incidence and outcome of organ failure. The purpose of this study is to establish the current incidence and mortality of organ failure in a homogenous population of critically ill trauma patients. METHODS All trauma patients admitted to the intensive care unit (ICU) at an urban Level I trauma center were prospectively studied. Patients were evaluated for the presence of organ failure using definitions proposed by Knaus and by Fry. Newer definitions of organ failure incorporating organ dysfunction and severity-of-illness scores were also obtained in all patients in an attempt to predict outcome. These included lung injury scores (acute respiratory distress syndrome scores), Acute Physiology and Chronic Health Evaluation (APACHE) II and III scores, Injury Severity Score (ISS), and multiple organ dysfunction scores. Primary outcomes assessed were death and the occurrence of organ failure by the various definitions. RESULTS Eight hundred sixty-nine trauma patients were admitted to the ICU and survived longer than 48 hours. Mean APACHE II and APACHE III scores at admission to the ICU and ISS were 12.2 +/- 22, 30.5 +/- 22.7, and 19 +/- 10, respectively. Single organ failure (SOF) occurred in 163 patients (18.7%) and multiple organ failure occurred in 44 patients (5.1%). All SOF was caused by respiratory failure. Respiratory failure occurred first in the majority of patients with multiple organ failure. Mortality was 4.3% with one organ system failure, 32% with two, 67% with three, and 90% when four organ systems failed. None of the patients with SOF died secondary to respiratory failure. Multiple stepwise regression analysis was performed to determine which of the following risk factors are associated with the occurrence of organ failure: mechanism of injury, lactate at 24 hours, ISS, APACHE II, APACHE III, acute respiratory distress syndrome score at admission, multiple organ dysfunction score at admission and total blood products transfused in 24 hours. Of these factors, APACHE III, lactate at 24 hours, and total blood products transfused in 24 hours were associated with the occurrence of organ failure. CONCLUSION The overall incidence of organ failure in a homogeneous trauma population appears to be lower than that reported in studies performed in heterogeneous patient populations in the 1980s. Mortality for SOF is low and appears to be related primarily to the patient's underlying injuries and not to organ failure. Mortality for two or three organ system failures is lower than reported 15 to 20 years ago. Mortality for patients with four or more organ system failures remains high, approaching 100%. PMID- 14566111 TI - Traumatic brain injury in U.S. Army paratroopers: prevalence and character. AB - BACKGROUND: This study presents self-reported traumatic brain injury (TBI) prevalence rates for 2,337 active duty U.S. Army soldiers who underwent baseline testing as part of a larger study of military TBI. METHODS: A computerized self report questionnaire was administered to a convenience sample of 2,337 highly functioning active-duty soldiers at Fort Bragg, North Carolina, who underwent baseline testing during a 13-month period in 1999 and 2000 as part of a larger ongoing Institutional Review Board-approved study examining the consequences of brain injuries among paratroopers. RESULTS: Approximately 23% of all of the soldiers surveyed reported sustaining a TBI after joining the Army. More than twice as many paratroopers reported sustaining TBI after joining the Army than did nonparatroopers (p < 0.001). Parachute-related TBI accounted for this difference. Nearly all of these injuries were mild. Less than 2% of paratroopers and no nonparatroopers reported loss of consciousness lasting more than 20 minutes. It was also shown that paratroopers with a history of TBI before joining the Army had a higher prevalence of TBI while serving in the Army (35%) than paratroopers without prior TBI (27.2%) (p = 0.002). CONCLUSION: This study demonstrates that parachuting appears to be a risk factor for mild TBI in the U.S. Army and that paratroopers with a history of TBI before joining the Army might be at somewhat increased risk of sustaining additional TBI while serving in the Army. PMID- 14566113 TI - Increased damage from rifle wounds of the chest caused by bullets striking commonly carried military equipment. AB - BACKGROUND: The purpose of this study was to investigate the fragmentation effect of the bullet that passes through the radio or ammunition magazine, which are essential pieces of equipment for a soldier, in thoracic gunshot injuries. METHODS: Twelve adult pigs were used. The pigs were separated into three groups (each group contained four pigs). The first group was shot without any obstacle between muzzle and subject. The second group was shot through a cartridge magazine placed in front of the subject's thoracic cage. The third group was shot with a radio in front of the subject's thoracic cage. RESULTS: It was observed that there was a large bullet entrance and fragmentation in the pigs that were shot with a radio or magazine placed over them, and there was no fragmentation in those that were shot without any obstacle. CONCLUSION: Metal equipment that soldiers carry causes bullet fragmentation. Fragmented bullets may cause excessive tissue disruption of intrathoracic vital organs. PMID- 14566114 TI - The pattern of gunshot injuries in a communal clash as seen in two Nigerian teaching hospitals. AB - BACKGROUND: The pattern of gunshot injury in the Ife-Modakeke communal clash between March 3 and May 23, 1999 was analyzed in this study. METHODS: The history was recorded for each patient presenting at the hospital with gunshot injuries from the clash. Each was examined and followed up through subsequent treatment in the wards and clinics. RESULTS: The study included 185 patients. The male-to female ratio was 22.1 to 1, and 45.9% of the patients ranged in age from 21 to 30 years. The mean age was 28.3 +/- 11.2 years, and the mortality rate was 8.6%. There were 120 warriors and 65 noncombatants. Government workers and students made up most of the noncombatants (42.5%), whereas drivers and farmers constituted the vast majority of the warriors (23.3%). Injury to the lower limbs was the most commonly seen injury (54.6%). Sustained fractures and femoral fractures were the most common fracture (n = 100, 54.1%). Head and colonic injuries were the greatest cause of mortality. CONCLUSION: For civilian gunshot injuries, morbidity and mortality rates are high in the less developed countries because of inadequate prehospital and hospital care. Efforts should be made to improve these facilities. PMID- 14566115 TI - Nonintracranial fatal firearm injuries in children: implications for treatment. AB - BACKGROUND: Understanding the injury characteristics of nonintracranial fatal (NIF) gunshot wounds in children treated in a statewide trauma system will help guide effective treatment strategies. METHODS: This study was a retrospective analysis of children fatally injured with firearms. The review included demographic information, firearm injury characteristics, and outcome. The setting included trauma centers participating in a statewide trauma registry. Patients were all children (age < 18 years) treated in trauma centers for NIF gunshot wounds from January 1988 through December 2000. The main outcome measures were characteristics of fatal firearm injuries in children. RESULTS: Over the 13-year period, there were 1,954 children with firearm injuries including 368 (18.8%) children with fatal wounds. Of the fatally wounded children, 177 (48.1%) had no intracranial injury. The NIF injury population was 90.4% male, with a mean age of 14.9 years (range, <1-17 years) and an Injury Severity Score of 38.2 (range, 9 75). Over 95% of deaths in this group occurred within 24 hours of admission. Although injuries to the thorax were most common (78.5%), 48.6% of the NIF cohort had injuries to multiple body regions, including 31% with injuries in both the abdomen and thorax. Compared with all children wounded by firearms, NIF firearm injury patients had, on average, more body regions injured (1.6 vs. 1.1, p < 0.001) and a greater total number of injuries (6.0 vs. 3.5, p < 0.001). Patients with an NIF injury were more likely to suffer a major vascular injury (54.8% vs. 13.8%, p < 0.001), lung injury (56.5% vs. 12.9%, p < 0.001), or cardiac injury (44.6% vs. 4.6%, p < 0.001) than all children with a firearm injury. CONCLUSION: Most children who arrive at trauma centers alive and subsequently die from NIF firearm injuries do so rapidly from major vascular and thoracic injury. Almost half of these children have injuries to multiple body regions, further complicating management. Innovative, aggressive treatment approaches should be sought to improve survival in this extremely injured cohort of children. PMID- 14566117 TI - Occult diaphragmatic injuries caused by stab wounds. AB - BACKGROUND: Missed diaphragmatic perforation caused by penetrating trauma can lead to subsequent strangulation of a hollow viscus, which has prompted the use of invasive diagnostic procedures to exclude occult diaphragmatic injuries in asymptomatic, high-risk patients. The objective of this study was to determine the incidence of occult diaphragmatic injuries caused by stab wounds of the lower chest and upper abdomen, and to examine the natural history and consequences of missed diaphragmatic injuries. METHODS: On the basis of patient data from two previous randomized studies from our institution, a retrospective analysis was performed on 97 patients treated for anterior stab wounds located between the nipple line, the umbilical level, and the posterior axillary lines not having indications for immediate surgical exploration. The patients were divided into two groups on the basis of their initial randomized management (open or laparoscopic exploration vs. expectant observation). RESULTS: In the exploration group (n = 47), four diaphragmatic injuries (9%) were detected (three left-sided and one right-sided). Excluding patients with associated injuries requiring surgical repair, the incidence of occult diaphragmatic injuries was 3 of 43 (7%). In the observation group (n = 50), there were two patients (4%) with delayed presentation of missed left-sided diaphragmatic injury 2 and 23 months later, respectively. Both injuries resulted from stab wounds of the left flank and presented with herniation of the stomach or small bowel and colon. The overall incidence of occult diaphragmatic injuries in left-sided thoracoabdominal stab wounds was 4 of 24 (17%), and was much lower after stab wounds of left epigastrium (0%), right lower chest (0%), and right epigastrium (4%). CONCLUSION: In asymptomatic patients with anterior or flank stab wounds of the lower chest or upper abdominal area, the risk of an occult diaphragmatic injury is approximately 7% which, if undetected, is associated with a high risk of subsequent hollow viscus herniation. Exclusion of an occult diaphragmatic injury with invasive diagnostic methods, such as laparoscopy or thoracoscopy, should be considered at least in left-sided stab wounds of the lower chest. PMID- 14566116 TI - Diagnostic laparoscopy in abdominal stab wounds: a prospective, randomized study. AB - BACKGROUND: The optimal strategy for identifying patients with abdominal stab wounds requiring surgical repair has not been defined. The potential benefits of diagnostic laparoscopy by incorporating it into the routine diagnostic workup of patients with anterior abdominal stab wounds was evaluated in a two-layer, randomized study. METHODS: From May 1997 through January 2002, stable patients without peritonitis but with demonstrated peritoneal violation were randomized (A) to exploratory laparotomy (AEL) (n = 23) or diagnostic laparoscopy (ADL) (n = 20). Simultaneously, patients with equivocal peritoneal violation on local wound exploration were randomized (B) to diagnostic laparoscopy (BDL) (n = 28) or expectant nonoperative management (BNOM) (n = 31). Hospital morbidity, length of stay, and costs were primary endpoints, with postdischarge disability being a secondary endpoint. RESULTS: In patients with peritoneal penetration (AEL vs. ADL), there were minimal differences in the therapeutic operation rate (8 of 23 [AEL] vs. 8 of 20 [ADL], p = 0.761), mortality (none), morbidity (3 of 23 vs. 2 of 20, p = 0.999), hospital stay (mean +/- SD) (5.7 +/- 2.5 vs. 5.1 +/- 4.0 days, p = 0.049), hospital costs (4.6 +/- 1.3 vs. 4.8 +/- 1.9 x 1,000 EUR, p = 0.576), and length of sick leave (34 +/- 12 vs. 29 +/- 11 days, p = 0.305). In patients with equivocal peritoneal penetration (BDL vs. BNOM), laparoscopy found more mostly minor organ injuries (7 of 28 [BDL] vs. 1 of 31 [BNOM], p = 0.022) with no significant difference in therapeutic operations (3 of 28 vs. 1 of 31, p = 0.337) or morbidity (3 of 28 vs. 0 of 31, p = 0.101), but was associated with increased length of stay (2.6 +/- 2.1 vs. 1.9 +/- 1.8 days, p = 0.022), hospital costs (4.2 +/- 1.3 vs. 1.5 +/- 1.1 x 1,000 EUR, p = 0.000), and sick leave requirements (18 of 23 vs. 8 of 28 of eligible patients, p = 0.001). CONCLUSION: In patients with demonstrated peritoneal violation, laparoscopy offers little benefit over exploratory laparotomy. In patients with equivocal peritoneal penetration on local wound exploration, laparoscopy detects more mostly minor organ injuries than expectant nonoperative management but is associated with increased hospital stay, costs, and sick leave requirements. Overall, diagnostic laparoscopy cannot be recommended as a routine diagnostic tool in anterolateral abdominal and thoracoabdominal stab wounds. PMID- 14566118 TI - Trauma assessment training with a patient simulator: a prospective, randomized study. AB - BACKGROUND: Patient simulators are computer-controlled mannequins that may increase realism during trauma training by providing real-time changes in vital signs and physical findings during trauma scenarios. We hypothesized that trauma assessment training on a patient simulator would be as effective as training with a more traditional moulage patient/actor. METHODS: This study was conducted during a surgery intern orientation at two academic trauma centers. Interns (n = 60) attended a basic trauma course, and were then randomized to trauma assessment practice sessions with either the patient simulator (n = 30) or a moulage patient (n = 30). After practice sessions, interns were randomized a second time to an individual trauma assessment test on either the simulator or the moulage patient. Two surgeon-judges rated each intern live and on video for completion of 50 predetermined assessment objectives (total score) divided into sections (primary and secondary survey, general performance, diagnostic studies/procedures, and plan) and the identification and management of an acute neurologic deterioration in the test patient (event score). Multiple linear regression with random student effects was used to estimate the independent effects of all study variables. RESULTS: Within randomized groups, mean trauma assessment test scores for all simulator-trained interns were higher when compared with all moulage-trained interns (71 +/- 8 vs. 66 +/- 8, respectively; p = 0.02). Simulator training independently showed a small but statistically significant improvement in both the total score and the event score (+4.6 and +8.6, respectively; p < 0.05). CONCLUSION: Use of a patient simulator to introduce trauma assessment training is feasible and compares favorably to training in a moulage setting. Continued research in this area of physician education is warranted. PMID- 14566119 TI - Are five-view plain films of the cervical spine unreliable? A prospective evaluation in blunt trauma patients with altered mental status. AB - OBJECTIVES: Clearing the cervical spine in a time-sensitive fashion is difficult. We hypothesized that admission computed tomographic scan of the occiput to T1 (CTS) with multiplanar reformatted images will replace five-view (odontoid, anteroposterior, lateral, and oblique) plain films of the cervical spine (CSX) in the initial evaluation of blunt trauma patients with altered mental status. METHODS: Between January and July 2001, all patients aged 16 years or older with altered mental status undergoing both CTS and CSX were prospectively entered into the study group. Attending physician interpretation defined the presence of cervical spine injury. Unstable fractures were defined as requiring surgical or halo stabilization. RESULTS: One thousand six patients met study criteria. One hundred sixteen patients had 172 cervical spine injuries (CSIs) (fracture and subluxation). CSX missed 90 of 172 (52.3%) CSIs in 65 of 172 (56.0%) patients. Anatomically, CSX failed to identify 14 of 15 occipital fractures (93.3%), 17 of 36 (47.2%) C1-3 fractures, and 59 of 121 (48.8%) C4-T1 CSIs. CSX failed to identify 5 of 29 (17.2%) patients with unstable CSIs. CTS failed to diagnose 3 of 172 (1.7%) CSIs that were stable (spinous process fractures at C6-7). Two patients exhibited spinal cord injury without radiologic abnormality missed by both modalities. CTS had a sensitivity of 97.4%, a specificity of 100%, a prevalence of 11.5%, a positive predictive value of 100%, and a negative predictive value of 99.7%. CSX had a sensitivity of 44.0%, a specificity of 100%, a prevalence of 11.5%, a positive predictive value of 100%, and a negative predictive value of 93.2%. CONCLUSION: CTS outperformed five-view CSX in a group of patients with altered mental status or distracting injuries. Five-view CSX failed to diagnose 52.3% of cervical spine fractures identified by CTS. Five-view CSX failed to diagnose five patients with unstable cervical fractures and failed to identify 93.3% of patients with occipital condyle fractures. PMID- 14566120 TI - Reformatted visceral protocol helical computed tomographic scanning allows conventional radiographs of the thoracic and lumbar spine to be eliminated in the evaluation of blunt trauma patients. AB - BACKGROUND: Patients suffering high-energy injuries are at risk for occult thoracic and lumbar spine fractures, and the standard of care includes radiographic spine screening. Most such patients require computed tomographic (CT) scanning to screen for chest and/or abdominal visceral injury. Helical CT (HCT) scanning represents a major technologic change that allows data to be reformatted after the patient has left the radiology suite. We explored the possibility of using reformatted visceral protocol HCT scanning to replace radiographs of the thoracic and lumbar spine in the evaluation of seriously injured patients. METHODS: A prospective evaluation of consecutive patients with thoracic and lumbar spine fractures admitted over a 12-month period to an urban Level I trauma center was completed. The ability of conventional radiography and reformatted HCT scanning to detect spine fractures was compared. RESULTS: Of 1,915 trauma patients admitted, 78 (4.1%), with an average Injury Severity Score of 21.3 +/- 1.2, sustained one or more thoracic (n = 35 patients) or lumbar (n = 43 patients) spine fractures. The sensitivity of reformatted HCT scanning as a screening test for spine fractures was 97% for thoracic and 95% for lumbar spine fractures, compared with a sensitivity of 62% for thoracic and 86% for lumbar conventional radiographs. CONCLUSION: Data obtained from HCT scanning performed to evaluate seriously injured multiple trauma patients for thoracic and abdominal visceral injury can be reformatted to screen for thoracic and lumbar spine fractures, providing accurate screening while eliminating the time, expense, and radiation exposure associated with conventional film radiography. PMID- 14566121 TI - Clinical predictors for the selective use of chest radiographs in pediatric blunt trauma evaluations. AB - BACKGROUND: Chest radiographs continue to be a routine part of the evaluation of children sustaining blunt trauma. This study sought to determine those clinical markers associated with an abnormal chest radiograph in nonintubated, pediatric, blunt trauma victims. METHODS: A retrospective case-control study was performed for severely injured pediatric trauma patients presenting to our emergency department between January 1, 1996, and December 31, 1997. Abnormal chest radiographs were identified through the trauma registry and four controls were matched to each case. Radiographs were reevaluated by our study radiologist. Variables associated with an abnormal chest radiograph were grouped to develop a set of clinical markers that could predict an abnormal chest radiograph with a high degree of sensitivity. RESULTS: An initial chest radiograph was obtained in 457 of 587 trauma patients. Thirty study patients with an abnormal radiograph that met inclusion criteria were analyzed with 133 controls. The presence of either an abnormal respiratory rate for age, chest tenderness, or back abrasions had a sensitivity of 1.0 (95% confidence interval, 0.86-1.0) and a specificity of 0.38 (95% confidence interval, 0.30-0.47). CONCLUSION: In pediatric trauma patients, the presence of chest tenderness, back abrasions, or an abnormal respiratory rate identified all abnormal chest radiographs. PMID- 14566122 TI - The roles of cyclic adenosine monophosphate- and cyclic guanosine monophosphate dependent protein kinase pathways in hydrogen peroxide-induced contractility of microvascular lung pericytes. AB - BACKGROUND: Sepsis and posttraumatic inflammatory processes are accompanied by definite changes in microvascular permeability, particularly in the lung. These permeability changes may occur because of damaged regulatory mechanisms at the level of the capillary wall. Pericytes are adventitial cells located within the basement membrane of capillaries. These cells contain multiple cytoplasmic processes that envelope endothelial cells, and are consequently thought to stabilize capillary walls and participate in microcirculation and endothelial cell permeability. Data from this laboratory and other laboratories have confirmed that pericytes are contractile cells, adding to the evidence that pericytes may influence or help regulate capillary permeability. We have already determined that hydrogen peroxide (H2O2) causes dose-dependent relaxation in microvascular lung pericytes (MLPs) at 10 minutes and, conversely, dose-dependent contraction at 30 minutes. It is the aim of this study to determine the mechanism of this biphasic contractile response. Specifically, we will determine whether cyclic adenosine monophosphate (cAMP)- or cyclic guanosine monophosphate (cGMP) dependent protein kinase intracellular pathways are responsible for the hydrogen peroxide-induced contractility of MLPs. METHODS: Rat MLPs were isolated by previously published protocol and cultured on collagen gel matrices. MLPs were pretreated with either ODQ, a soluble guanylate cyclase inhibitor (100 mumol/L), for 15 minutes; GKIP, a protein kinase G inhibitor (100 mumol/L), for 1 hour; SQ22536, an adenylate cyclase inhibitor (100 mumol/L), for 15 minutes; or H89, a protein kinase A inhibitor (10 mumol/L), for 1 hour. Hydrogen peroxide was then introduced to each MLP culture at 10 mumol/L, 100 mumol/L, and 1 mmol/L. After each of these treatments, the surface area of the collagen gels was digitally quantified at 10 and 30 minutes. RESULTS: SQ22536 attenuated both relaxation at 10 minutes and the contraction seen at 30 minutes for all concentrations of H2O2. H89 caused a marked basal relaxation and prevented the cells from contracting at 30-minute exposures to all concentrations of H2O2. Both ODQ and GKIP attenuated the relaxation at 10 minutes but had no affect on the later contraction. CONCLUSION: The cGMP-dependent protein kinase pathway is a mechanism for H2O2 induced relaxation of MLPs. Up-regulation of cAMP and cGMP is responsible for early H2O2-induced relaxation and late contraction. Protein kinase A (cAMP dependent protein kinase pathway) may be an important intracellular signaling protein in the H2O2-induced contraction of MLPs or may be unable to down-regulate cAMP once inhibited. This evidence further supports the concept that there are separate intracellular pathways that regulate divergent cellular responses. This idea parallels the clinical concept of reversible and irreversible dysfunction of cellular processes in shock, and that the cellular dysfunction is initiated by separate intracellular pathways. PMID- 14566123 TI - Fusion of the septic ankle: experience with 15 cases using hybrid external fixation. AB - BACKGROUND: In cases of septic joint destruction, an unfavorable situation of soft tissues and chronic osteomyelitis are responsible for high failure rates of ankle fusions. We wanted to evaluate the control of infection and the fusion rate using hybrid external fixators for the fusion of the septic ankle in a prospective study. METHODS: From 1996 to 1998, 15 arthrodeses were performed using hybrid external fixators. All patients had a combination of bone and soft tissue infections. Fourteen patients suffered from sequelae of posttraumatic osteoarthritis, and one patient suffered from rheumatoid arthritis. In 14 patients, pathogens could be identified; in 87%, Staphylococcus aureus was found. Eight patients had relevant concomitant diseases. RESULTS: The preservation of limbs by solid tibiotalar fusion was achieved in 14 patients (93%). One patient maintained an infected pseudarthrosis. During the 12-month follow-up, three patients had a fistula that persisted, with two patients having a solid arthrodesis. Full weight-bearing was possible for all the patients with a successful fusion. Seventy-five percent of the patients that had not retired at the time of the study regained their fitness for work. CONCLUSION: The hybrid external fixator presents a successful alternative for those arthrodeses of ankle joints where complications such as bone/joint infections or poor soft tissue conditions occur. PMID- 14566124 TI - Identification of the optimal intercondylar starting point for retrograde femoral nailing: an anatomic study. AB - BACKGROUND: Retrograde nailing of femoral shaft fractures is an effective and increasingly more popular method of fracture fixation. However, concern remains regarding the effect of the intercondylar entry-portal location on knee function. METHODS: The optimal entry-portal location was identified in cadaver femurs. Approximating the clinical intraoperative situation, a threaded guidewire was inserted into each of 26 distal femur specimens and positioned in the center of the femoral shaft as determined by anteroposterior and lateral fluoroscopic imaging. Each guidewire was then overdrilled with a 12-mm cannulated drill bit. All entry-portal locations were recorded relative to the posterior cruciate ligament attachment and the intercondylar groove and mapped relative to the known patellofemoral contact area. RESULTS: The starting holes averaged 6.21 mm anterior to the posterior cruciate ligament attachment and 2.67 mm medial to the intercondylar groove. Overall, 100% of starting portals were located in safe areas relative to the patellofemoral contact area. CONCLUSION: In the vast majority of femurs, the optimal entry portal for retrograde femoral nailing (in line with the long axis of the femur) is located in the expected safe position, anterior to the posterior cruciate ligament insertion and slightly medial to center of the intercondylar groove. However, because of anatomic variability, the ideal starting position occasionally may be located in a patellofemoral contact area. Potential compromise of the patellofemoral contact area by the retrograde nail entry portal can and should be recognized before nailing, allowing the surgeon the option of altering the surgical technique. PMID- 14566125 TI - Predictors of death in patients with life-threatening pelvic hemorrhage after successful transcatheter arterial embolization. AB - OBJECTIVE: The purpose of this study was to determine predictors of death in patients with pelvic fracture whose pelvic arterial hemorrhage is controlled successfully by transcatheter arterial embolization (TAE). METHODS: From January 1996 to December 2000, 61 patients with a pelvic fracture who had pelvic arterial hemorrhage were treated at our Level I trauma center according to a protocol that assigns a high priority to diagnostic and therapeutic angiography within the algorithm. Angiography is performed before laparotomy in patients with hemoperitoneum, who can be stabilized by fluid resuscitation, and otherwise afterward. External fixation was performed immediately after TAE in the angiography suite. Predictors of outcome were determined retrospectively by univariate and multivariate analysis using anatomic and physiologic parameters. RESULTS: Forty-eight patients survived and 13 died. TAE successfully controlled pelvic arterial hemorrhage in all patients. Predictors of death included posterior pelvic arterial injury and an elevated Acute Physiology and Chronic Health Evaluation II score (odds ratio, 15.6 and 23.9, respectively). Need for fluid requirements to achieve hemodynamic stability were higher in nonsurvivors than in survivors. Outcome did not correlate with the type of fracture or the Injury Severity Score. CONCLUSION: Application of angiography as a therapeutic intervention in patients with pelvic arterial bleeding may reduce the need for surgery, thereby avoiding or minimizing this additional trauma. PMID- 14566127 TI - The use of free tissue transfers in acute thermal and electrical extremity injuries. AB - BACKGROUND: This article presents a retrospective analysis of a case series of patients requiring free tissue transfers for acute thermal and electrical injuries in a single burn center. METHODS: Eight patients, in the period between 1997 and 2001, were referred for evaluation of extensive thermal injury to one or more extremities, which required free tissue transfer to achieve coverage. A total of 12 flaps were performed on eight patients. Data including flap type, flap viability, complications, hospital stays, length of rehabilitation, and time until return to work were collected. RESULTS: Twelve flaps were performed on eight patients. One patient died. We report a flap failure rate of 9% (1 of 12) and two postoperative infections. Follow-up ranged from 3 months to 4 years. Flap types included lateral arm, radial forearm, rectus abdominis, posterior lateral thigh, and omentum and temporoparietal fascia. Mean hospital stay was 22 days, with a mean rehabilitation time of 9.7 months. Sixty-two percent of patients returned to work. CONCLUSION: Free tissue transfer is a safe and efficacious method of early surgical management of thermal and electrical burns. Electrical burns account for a disproportionate number of injuries requiring free tissue transfers. These injuries frequently necessitate the use of multiple free flaps. Postoperative complications and length of stay can be minimized by the use of split donor flaps during one operative session. PMID- 14566126 TI - The clinical correlation of temporal bone fractures and spiral computed tomographic scan: a prospective and consecutive study at a level I trauma center. AB - BACKGROUND: Undetected temporal bone fractures (TBFs) can lead to complications such as hearing loss, facial nerve paralysis, otorrhea, or otorhinorrhea, and can be the cause of life-threatening bacterial meningitis and can easily be missed. METHODS: We prospectively studied patients with head trauma between January 2000 and January 2001. All patients were examined by the attending trauma physician and then underwent cranial helical computed tomographic (CT) scan independent of clinical findings and Glasgow Coma Scale (GCS) score to determine the proportion of TBFs that would have been missed if diagnosis had been based only on clinical evaluation. RESULTS: Three-hundred fifty consecutive patients with head trauma were studied during the 12-month period (GCS score: range, 3-15; median, 13). In 34 (9.7%) of these patients, 38 TBF (30 unilateral and 4 bilateral) were diagnosed by CT scan. However, clinical signs of TBF were present in only 22 of the 34 patients (65%). All missed TBFs were unilateral. Of the patients with missed TBFs, 8 of 20 (40%) had GCS scores of 14 to 15, 1 of 8 (12.5%) had a GCS score of 9 to 13, and 3 of 6 (50%) had GCS scores < 9. Four of 34 (12%) patients developed clinical complications. CONCLUSION: TBFs are common injuries in patients with head trauma. More than one third of these fractures may be missed by clinical diagnosis alone. Although the clinical importance of the missed TBF is debatable, 12% of our patients developed complications. Therefore, to rule out these lesions, routine cranial helical CT scan should be recommended in all patients presenting with head trauma, independent of clinical findings and GCS score. PMID- 14566128 TI - Paramedic-administered neuromuscular blockade improves prehospital intubation success in severely head-injured patients. AB - BACKGROUND: The purpose of this study was to evaluate the effect of paramedic administered neuromuscular blocking agents as part of a rapid-sequence intubation (RSI) protocol on successful intubation of severely head-injured patients in a large, urban prehospital system. METHODS: Adult head-injured patients were prospectively enrolled over 1 year using these inclusion criteria: Glasgow Coma Scale (GCS) score of 3 to 8, transport time > 10 minutes, and inability to intubate without RSI. Midazolam and succinylcholine were administered before laryngoscopy; rocuronium was given after tube placement was confirmed using capnometry, syringe aspiration, and pulse oximetry. The Combitube was used as a salvage airway device. All adult trauma victims with a GCS score of 3 to 8 were identified during the first 12 months of the study as the trial cohort and from the preceding 12 months as the control cohort. The trial and control cohorts were compared with regard to demographic data, mechanism of injury, initial vital signs, and GCS scores. The primary outcome measure was intubation success, defined as insertion of either an endotracheal tube or a Combitube, with patients stratified by GCS score. RESULTS: The trial cohort (n = 249) and control cohort (n = 189) were similar with regard to demographic data, mechanism of injury, and initial vital signs and GCS scores. Intubation success rates increased significantly during the trial period for all patients and when stratified into GCS score of 3 and GCS score of 4 to 8. The percentage of patients intubated without neuromuscular blocking agents actually increased during the trial period. Although the number of intubations by helicopter flight crews decreased during the trial, the overall use of aeromedical resources did not change. CONCLUSION: Paramedic-administered neuromuscular blockade as part of an RSI protocol improves intubation success in a large, urban prehospital system. PMID- 14566129 TI - The esophageal Doppler monitor in mechanically ventilated surgical patients: does it work? AB - BACKGROUND: Assessment of cardiac volumes and cardiac output (CO) using a pulmonary artery catheter (PAC) in mechanically ventilated patients can be inconsistent and difficult. The esophageal Doppler monitor (EDM) is emerging as a potential alternative to the PAC. This prospective study evaluated the comparative accuracy between the PAC and EDM for preload assessment and CO in mechanically ventilated surgical patients. METHODS The EDM was placed in 15 patients with PACs in place. A total of 187 simultaneously measured EDM and PAC comparative data sets were obtained. The Pearson correlation (r) was used to compare measurements, with significance defined as a value of p < 0.05. RESULTS: CO measured by EDM and PAC correlated closely (r = 0.97, p < 0.0001). Corrected flow time (FTc), a measure of left ventricular filling, correlated with PAC CO to the same degree as pulmonary capillary wedge pressure (PCWP) when positive end expiratory pressure (PEEP) was < 10 cm H2O (FTc, r = 0.51; PCWP, r = 0.56). When PEEP was > or = 10 cm H2O, FTc correlated with PAC CO better than PCWP (FTc, r = 0.85; PCWP, r = 0.29). CONCLUSION: FTc correlates with EDM and PAC CO better than PCWP. On the basis of the current study, it is reasonable to conclude that the EDM is a valuable adjunct technology for CO and preload assessment in surgical patients on mechanical ventilation, regardless of the level of mechanical ventilatory support. PMID- 14566130 TI - Factors in intestinal lymph after shock increase neutrophil adhesion molecule expression and pulmonary leukosequestration. AB - BACKGROUND: Because the ischemic gut may produce factors that initiate systemic inflammation, we tested the hypothesis that factors released from the gut into the mesenteric lymphatics increase neutrophil (PMN) adhesion molecule expression after trauma and shock. METHODS: At 1 and 4 hours after hemorrhagic shock (30 mm Hg x 90 minutes) plus trauma (laparotomy) (T/HS) or sham-shock (T/SS), with or without mesenteric lymph duct ligation, PMN CD11b and CD18 expression was assessed in male rats. In additional rats, mesenteric lymph samples were tested for their ability to increase PMN CD11b expression in vitro. Lastly, at 4 hours after T/SS or T/HS with or without lymph duct ligation, pulmonary PMN sequestration was measured. RESULTS: Compared with T/SS rats, T/HS was associated with up-regulation of PMN CD11b and CD18 expression, which was largely prevented by ligation of the mesenteric lymph duct (p < 0.01). Lymph duct ligation also prevented T/HS-induced pulmonary leukocyte sequestration (p < 0.01). In addition, mesenteric lymph from rats subjected to T/HS but not T/SS increased CD11b expression (p < 0.01). CONCLUSION: Factors produced or released by the postischemic intestine and carried in the mesenteric lymph appear responsible for PMN activation and pulmonary PMN sequestration after an episode of T/HS. PMID- 14566131 TI - Intravascular volume depletion in a 24-hour porcine model of intra-abdominal hypertension. AB - BACKGROUND: The purpose of the study was to examine hemodynamic parameters and intravascular volume in a porcine model in the presence of intra-abdominal hypertension (IAH) lasting for 24 hours. METHODS: Twelve pigs (52.5 +/- 4.9 kg) were studied over a period of 24 hours. In six animals, the intra-abdominal pressure was increased to 30 mm Hg via carbon dioxide-pneumoperitoneum. The others served as controls. Using the double-indicator dilution technique, intrathoracic blood volume (ITBV), total circulating blood volume, and cardiac output (CO) were measured. Standard parameters (e.g., central venous pressure [CVP]), were also recorded. RESULTS: In the presence of IAH, ITBV and total circulating blood volume were significantly reduced to 55% and 67% of control values. CO decreased to 27% and CVP increased fourfold. CONCLUSION: IAH leads to significant intravascular volume depletion that is not reflected by the CVP. Assessment of CO and ITBV in the presence of a critically increased intra abdominal pressure is therefore recommended. PMID- 14566132 TI - A randomized, controlled trial comparing arteriovenous to venovenous rewarming of severe hypothermia in a porcine model. AB - BACKGROUND: The purpose of this study was to evaluate active rewarming using continuous arteriovenous rewarming (CAVR) and continuous venovenous rewarming (CVVR) methods during severe hypothermia using an electromagnetic fluid warmer. Rapid rewarming using these techniques is superior to passive rewarming and is possible with commercially available equipment. METHODS: Eighteen swine (55-65 kg) were assigned to CAVR, CVVR, or control. Vascular access was obtained via central lines (8.5-French) in all subjects. Subjects were cooled to 27 degrees C (80.6 degrees F) in an ice bath, and then dried, covered, and connected to the rewarming device. The carotid artery (CAVR) or internal jugular vein (CVVR) was used for circuit inflow. Warmed 39 degrees C (102.2 degrees F) blood was returned via the femoral vein. Hemodynamic parameters and temperatures (pulmonary artery and rectal) were recorded until reaching an endpoint of a pulmonary artery temperature of 37 degrees C (98.6 degrees F). RESULTS: Mean rewarming time in the CAVR group was 2 hours 14 minutes, with a mean rewarming rate of 4.5 degrees C/h (8.1 degrees F/h, 0.034 degrees C/kg/h). Total circulating volume averaged 65 L. CVVR averaged 3 hours 8 minutes, with a mean rewarming rate of 3.2 degrees C/h (5.8 degrees F/h, 0.024 degrees C/kg/h). Total circulating volume averaged 67 L. Controls averaged 10 hours 42 minutes, with a mean rate of 0.9 degrees C/h (1.7 degrees F/h, 0.007 degrees C/kg/h). The CAVR group was faster than the CVVR group in both the rewarming rate and total time to rewarming (p = 0.034 and p = 0.040, respectively). Both experimental groups were significantly different from controls in rewarming rate and total time to rewarming (p < 0.001). CONCLUSION: CAVR offers the most rapid rate of rewarming. CVVR offers a rapid rate using less invasive procedures. Both techniques are markedly superior to passive rewarming methods typically used during early resuscitation. PMID- 14566134 TI - The effect of the bovine hemoglobin oxygen therapeutic HBOC-201 on human neutrophil activation in vitro. AB - BACKGROUND: We previously demonstrated that HBOC-201 is an efficient resuscitation fluid. However, little is known about its immunomodulatory effects. The goal of this study was to investigate human neutrophil activation after exposure to HBOC-201 and other low-volume resuscitation fluids. METHODS: Whole blood from 10 healthy volunteers was serially diluted with HBOC-201, 7.5% hypertonic saline (HTS), 7.5% hypertonic saline-6% dextran 70 (HSD), Hextend, or PentaLyte. After an incubation period, samples were analyzed for oxidative burst and CD11b expression using flow cytometry. RESULTS: Increases in both CD11b and oxidative burst activity were noted in samples diluted with HBOC-201, Hextend, PentaLyte, and lactated Ringer's solution. Samples maximally diluted with HBOC 201 demonstrated increased CD11b expression when compared with lactated Ringer's solution and Hextend (196.11% vs. 22.68% and 33.97%; p = 0.018 and p = 0.033). CONCLUSION: Although differences were noted only at the highest dilutions, further studies investigating the effects of HBOC-201 are warranted. PMID- 14566133 TI - A comparison of the hemoglobin-based oxygen carrier HBOC-201 to other low-volume resuscitation fluids in a model of controlled hemorrhagic shock. AB - BACKGROUND: The ideal resuscitation fluid for military applications would be effective at low volumes, thereby reducing logistical constraints. We have previously shown that the bovine hemoglobin-based oxygen carrier HBOC-201 is an effective low-volume resuscitation fluid. The goal of this experiment was to evaluate the effectiveness of HBOC-201 in comparison with other low-volume resuscitation fluids in a swine model of controlled hemorrhagic shock. METHODS: Forty-two immature female Yorkshire swine (55-70 kg) were divided into seven groups of six. Animals were hemorrhaged to a mean arterial pressure of 30 mm Hg. After 45 minutes, animals were resuscitated to a mean arterial pressure of 60 mm Hg with one of the following agents: hypertonic saline 7.5% (HTS), hypertonic saline 7.5%/Dextran-70 6% (HSD), pentastarch 6%, hetastarch 6%, or HBOC-201. Lactated Ringer's (LR) solution was used as a standard resuscitation control. Another group of animals received no resuscitation. Resuscitation was continued for 4 hours. Hemodynamic variables and oxygen consumption were measured continuously. Arterial and mixed venous blood gases and serum lactate levels were measured at intervals throughout the experiment. Data were analyzed using analysis of variance with Tukey's post hoc test when appropriate. Significance was defined as p < 0.05. RESULTS: Five of six animals in the no-resuscitation control group, six of six in the HTS group, and one animal in the HSD group died before completion of the study. All other animals survived to completion. Animals receiving resuscitation with HBOC-201 had significantly lower cardiac output, mixed venous oxygen saturation levels, and urinary output throughout the resuscitation period; however, there were no differences with regard to lactate, base excess, or oxygen consumption. Animals receiving HBOC-201 required significantly less fluid than any other group. CONCLUSION: In this model, hypotensive resuscitation with HBOC-201 restores tissue oxygenation and reverses anaerobic metabolism at significantly lower volumes when compared with HTS, HSD, pentastarch, or hetastarch solutions. These data suggest that HBOC-201 would be an effective primary resuscitation fluid for far-forward military or rural trauma settings where logistic constraints and prolonged transport times are common. However, when HBOC-201 is administered as a primary resuscitation fluid in hypotensive protocols, common clinical markers for determining adequacy of resuscitation may not be useful. PMID- 14566135 TI - Reducing trauma payment denials with computerized collaborative billing. AB - BACKGROUND: Trauma care demands constant physician availability, resulting in rotational coverage systems. Third-party payors consider separate trauma surgeon bills as originating from the same individual. Trauma surgeons may be unaware of their colleagues' billing history on jointly managed patients. Not all postoperative procedures and evaluation and management services are denied by global surgical package rules. We investigated whether a networked billing program designed to crosscheck for global package coding concerns would reduce payment denials. METHODS: A networked relational database was created for trauma surgeons to enter billable encounters, displaying global periods and operative diagnoses while prompting for postoperative modifiers. Denials were compared for equivalent time periods before and after program initiation. RESULTS: Payment denials fell from 361 to 16 for "bundled" evaluation and management services and from 55 to 13 for bundled postoperative procedures. Time spent on billing decreased and legibility improved. Overall savings totaled $183,404. CONCLUSION: Collaborative billing can improve payments for professional trauma care. PMID- 14566136 TI - Clostridium difficile pericarditis complicating pseudomembranous colitis in a trauma patient. PMID- 14566137 TI - Functional reconstruction of a massive defect in the two forearm bones. PMID- 14566138 TI - Endovascular repair of a traumatic pseudoaneurysm of the thoracic aorta in a patient with concomitant intracranial and intra-abdominal injuries. PMID- 14566139 TI - A case of in utero fetal brain trauma after motor vehicle collision. PMID- 14566140 TI - Tibial fracture as a complication of intraosseous infusion during pediatric resuscitation. PMID- 14566141 TI - Leg amputation after intraosseous infusion in a 7-month-old infant: a case report. PMID- 14566142 TI - Removal of unusual neglected foreign bodies from the urinary bladder: case report. PMID- 14566143 TI - Penile refracture: case report. PMID- 14566144 TI - Complete occlusion after blunt injury to the abdominal aorta. AB - BACKGROUND: Injury to the abdominal aorta after blunt trauma is uncommon. When this injury results in complete vessel occlusion, the presentation is dramatic. Timely intervention is essential. METHODS: After a case report, we examined all reported cases of complete occlusion after blunt injury to the abdominal aorta and reviewed the cause, presentation, and management of this injury. RESULTS: Complete vessel occlusion arises from intimal injury. The most frequent mechanism is compression from a seat belt or steering wheel during a motor vehicle crash. Patients present with absent femoral and distal pulses in association with lower extremity neuropathy. Intervention commonly involves bypass grafting of the abdominal aorta. CONCLUSION: Complete occlusion after blunt trauma to the abdominal aorta is rare. Neurologic deficits most commonly arise from peripheral nerve ischemia. Reperfusion within 6 hours confers a greater chance of limb salvage and neurologic recovery. PMID- 14566145 TI - The effect of arm traction on the adequacy of cross-table lateral cervical spine radiographs. PMID- 14566146 TI - Duodenal trauma. PMID- 14566147 TI - Small bowel in hypovolemic shock appearing calamari-like. PMID- 14566148 TI - Reorganization of gait after limb-saving surgery of the lower limb. AB - OBJECTIVE: In this study, the concept of a cognitive dual-task performance and visual restriction during walking has been used to study the recovery of gait after limb-saving surgery in ten patients. DESIGN: All patients were recovering from some form of treatment to tumors of the lower limbs. Patients had to walk on a treadmill at their preferred speed. During the course of recovery, we measured normal walking, walking while performing an attention-demanding dual task, and walking during restricted vision, starting 5 mo postoperatively. RESULTS: Patients are able to reach an acceptable level of gait within 15 mo, especially when the basic locomotor activity (i.e., step-cycle duration, walking speed, gait symmetry) is taken into account. Nevertheless, the results showed that during the recovery, the patients were still hindered by the dual task and visual restriction while walking because they exhibited a decrease in step-cycle duration under these conditions. CONCLUSIONS: In general, an improvement in walking speed and a decrease in asymmetry was seen. On the other hand, patients still had a basically reduced control of gait after the 15-mo recovery period. This can be attributed to a lack of gait automatism caused by an irreversible loss of somatosensory input. PMID- 14566149 TI - Temporospatial parameters of gait after obturator neurolysis in patients with spasticity. AB - OBJECTIVE: The management of spasticity should be implemented with the most appropriate pharmacologic agents. Ideally, these agents should provide functional improvement with minimal adverse effects. The aim of this study was to evaluate the width of the base of support and the velocity of gait before and after obturator nerve block. Blocks were performed with aqueous phenol solution in patients with unilateral hip adductor muscle overactivity that resulted in scissoring gait. The goal was to analyze functional improvement and quantify outcomes and to attempt to document adverse effects. DESIGN: This retrospective study analyzed data from 24 patients' files. Inclusion criteria included subjects whose main functional complaint was an adducted gait pattern. All subjects were able to consent for and to undergo unilateral obturator neurolysis with 7% phenol solution. Temporospatial parameters of gait were obtained using the Gait Mat II before and after obturator nerve injection. RESULTS: The analysis showed no statistical change in the walking velocity or step length. However, the width of the base of support was significantly increased after injection. No postblock complications were reported. CONCLUSION: Obturator neurolysis with 7% phenol solutions is an effective procedure to decrease hip adductor muscle overactivity without reported complications. In the studied population, improvement was found in the width of the base of support without immediate change in walking velocity or step length. PMID- 14566150 TI - Placement of a plate under the forefoot in stance: decreasing the excitability of the soleus motoneuron pool. AB - OBJECTIVE: To investigate the effect of the placement of a plate under the sole of the forefoot on the excitability of the soleus motoneuron pool in stance. DESIGN: While eight neurologically intact subjects remained in stance, a square plate (30 x 30 mm), either 3 mm or 6 mm in thickness, was placed under the left medial plantar eminence, under the left lateral plantar eminence, or under the left forefoot sole between the eminences, or a rectangular plate (120 x 30 mm), either 3 mm or 6 mm in thickness, was placed under the left metatarsal heads. To investigate the excitability of the soleus motoneuron pool, the Hmax/Mmax ratio was estimated. RESULTS: Analysis of variance revealed that the region under which the plate was placed had a significant effect and that the thickness of the plate had a nonsignificant effect on the Hmax/Mmax ratio, without significant interaction between the two factors. Post hoc analysis revealed a significant decrease in the Hmax/Mmax ratio when the square plate was placed under the lateral plantar eminence. CONCLUSIONS: A square plate placed under the lateral plantar eminence inhibited the excitability of the ipsilateral soleus motoneuron pool in stance. This intervention may be useful for reducing spasticity. PMID- 14566151 TI - Measurement of forefoot varus angle by laser technology in people with flexible flatfoot. AB - OBJECTIVE: The purpose of this study was to measure the forefoot varus angles in subjects with and without flexible flatfoot (FF) by using laser foot-scanning technology. DESIGN: In the rehabilitation laboratory of a tertiary medical center, 70 subjects with FF, ranging in age from 10 to 50 yr, were recruited. The control group consisted of 30 volunteers without clinical evidence of FF. A total of 100 positive casts were obtained by having their subtalar joints kept in a neutral position. The plantar surface of the positive cast was scanned by a Yeti 3D Foot Scanner. A straight line was drawn between the first and fifth metatarsophalangeal joints. The forefoot varus angle was measured from this line in relation to the line parallel to the ground. RESULTS: The mean forefoot varus angle was calculated to be 5.01 +/- 4.51 degrees in our FF subjects, and 1.23 +/- 1.96 degrees in the control group. Significant statistical difference in forefoot varus angle was noted between these two groups. CONCLUSIONS: The laser foot scanning technique offers fast and accurate measurement of the forefoot varus angles. An average forefoot varus angle of approximately 5 degrees was observed in subjects with FF, which was significantly greater than the subjects without FF. Therefore, we recommend the concept of incorporating adequate forefoot posting at the medial forefoot area of an insole in treating people with FF. PMID- 14566152 TI - Effectiveness of the five-handle position grip strength test in detecting sincerity of effort in men and women. AB - OBJECTIVE: The five-handle position (five-rung) test is used to determine sincerity of effort of grip strength. However, there is a controversy in the literature concerning its validity and effectiveness. The purpose of the study was to investigate whether test results are affected by the amount of strength exerted by the gripping hand and to determine the test's effectiveness. DESIGN: A total of 30 hand therapy patients performed the five-rung grip test both maximally and submaximally with both the injured and uninjured hands. The standard deviation across the five strength trials was used to measure the shape of the curve. Sensitivity and specificity values were calculated for each sex. RESULTS: The repeated measures analyses of variance revealed that average strength and the standard deviation were greater for men than for women, for maximal effort than for submaximal effort, and for the injured hand in comparison with the healthy hand. The most optimal standard deviation cutoff value was 8.5 (sensitivity, 0.70; specificity, 0.83), and the proportional area under the receiver operator characteristic curve was greater for the men (89%) than for women (80%). CONCLUSIONS: The shape of the curve generated by the five-rung test was strength dependent; thus, the test may yield biased results when assessing sincerity of effort in people with weakened hands. The five-rung test was less effective for women than for men. Thus, we recommend that the five-rung test not be used to detect sincerity of effort. PMID- 14566153 TI - Electromyographic biofeedback in the treatment of the hemiplegic hand: a placebo controlled study. AB - OBJECTIVE: To evaluate the efficacy of electromyographic (EMG) biofeedback treatment in the functional recovery of the hemiplegic hand. DESIGN: A total of 27 patients were randomly assigned to EMG biofeedback or placebo EMG biofeedback groups. Both treatments were applied five times a week for a period of 20 days. In addition, the patients in both groups received an exercise program according to the Brunnstrom's neurophysiologic approach. Goniometric measurements for wrist extension, scale for judging the performance of drinking from a glass, Brunnstrom's stages of recovery for hand, and surface EMG potentials were used for the clinical assessments. All patients were assessed before treatment and after 20 treatment sessions. RESULTS: The results showed that there were statistically significant improvements in all variables in both groups, but the improvements in active range of motion and surface EMG potentials were significantly greater in the EMG biofeedback group at the end of the treatment. CONCLUSION: Our study demonstrates the potential benefits of EMG biofeedback in conjunction with neurophysiologic rehabilitation technique to maximize the hand function in hemiplegic patients. PMID- 14566154 TI - Muscular atrophy in the hemiplegic thigh in patients after stroke. AB - OBJECTIVES: This study evaluated muscular atrophy in the hemiplegic limbs by assessing the muscle volume of the thighs in stroke patients. DESIGN: Muscle volume of the bilateral thighs was determined by computed tomographic scanning in 50 hemiplegic patients after stroke. RESULTS: The average muscle volume in the hemiplegic side was significantly lower than that in the nonhemiplegic side. When the patients were divided into the two groups aged <65 yr old and >/=65 yr, age dependent reduction in the muscle volume was significant only in the nonhemiplegic side. In addition, the ratio of the muscle volume in the hemiplegic side to that in the nonhemiplegic side was significantly lower in the older group than in the younger group. Muscle volume in both hemiplegic and nonhemiplegic sides correlated positively with Barthel index and negatively with patient age. CONCLUSION: Muscle volume decreases in the hemiplegic side in stroke patients. PMID- 14566155 TI - Nocturnal oximetry: utility in the respiratory management of amyotrophic lateral sclerosis. AB - OBJECTIVE: Current recommendations are to institute nocturnal nasal ventilation for amyotrophic lateral sclerosis patients with a forced vital capacity (FVC) of <50% of predicted normal. The purpose of this study was to determine whether this is appropriate. DESIGN: A total of 87 nocturnal oximetry evaluations were performed on 78 consecutive amyotrophic lateral sclerosis patients symptomatic for sleep-disordered breathing. Nocturnal oximetry measurements were compared for those with FVC >50% vs. those with FVC of <50% of normal. FVC was measured sitting and supine. RESULTS: A considerable number of these symptomatic patients manifested evidence of nocturnal hypoxemia as measured by oximetry. However, there was no significant difference between patients with sitting percentage predicted FVC above and below 50% predicted in minimum oxygen saturation, mean oxygen saturation, percentage of time spent with oxygen saturation of <88%, and number of events per hour. There was no significant difference between patients with supine percentage-predicted FVC above and below 50% predicted in minimum oxygen saturation, mean oxygen saturation, percentage of time spent with oxygen saturation of <88%, and number of events per hour. CONCLUSION: The recommendation that FVC be <50% of normal is inappropriate for justifying introduction of nocturnal nasal ventilation. Many patients are symptomatic at higher FVC and manifest evidence of nocturnal hypoxemia. Nocturnal oximetry adds additional practical information for justifying earlier respiratory intervention for symptomatic patients. PMID- 14566156 TI - Haloperidol, but not olanzapine, impairs cognitive performance after traumatic brain injury in rats. AB - OBJECTIVE: Traumatic brain injury can cause a variety of impairments, including persistent alterations in personality, mood, and cognition. Antipsychotic agents are frequently used to treat pathologic behaviors in traumatic brain injury patients, but the influence of prolonged administration of such drugs on cognition after injury is unknown. The effects of two antipsychotic drugs on cognitive recovery after traumatic brain injury were assessed using the fluid percussion model in rats. DESIGN: The typical antipsychotic, haloperidol, and the third-generation antipsychotic, olanzapine, were administered via intraperitoneal injection beginning 24 hr after injury and continuing daily for the duration of the study. Morris water maze performance was assessed on days 11-15 postinjury. RESULTS: Haloperidol, an antagonist acting on D2-like dopamine receptors, exacerbated the cognitive deficits induced by injury, as injured rats treated with 0.30 mg/kg haloperidol performed worse in the Morris water maze than injured rats treated with vehicle. CONCLUSIONS: Our results demonstrate the importance of the D2 receptor in cognitive recovery after traumatic brain injury. Also, the data illustrate that some classes of antipsychotic drugs may influence cognitive recovery, and further research is needed to determine the optimal pharmacologic treatment of aggression, agitation, and other pathologic behaviors in patients with traumatic brain injury. PMID- 14566157 TI - Assessment of wheelchair drag resistance using a coasting deceleration technique. AB - OBJECTIVE: To apply a recently developed coasting deceleration method to measure rolling and aerodynamic resistances opposing wheelchair propulsion on a variety of different wheelchairs and wheel combinations and on two different ground surfaces. DESIGN: For each condition, 20-25 trials were performed across a speed range of approximately 70-300 m/min. The least-squares method was then used to arrive at values for the coefficient of rolling resistance (CR) and effective frontal area of the wheelchair and occupant. RESULTS: Wheelchair rolling resistance was found to be velocity dependent under some circumstances. CR values on linoleum differed among folding lightweight wheelchairs and when compared with a rigid ultralight and racing wheelchair. Changing rear wheels and tires on one wheelchair resulted in a 14% difference in CR. Carpet increased CR values by an average of 0.0118 over the values determined on linoleum. As expected, effective frontal area of the wheelchair and occupant values were lower for the racer than for the folding lightweight wheelchair. CONCLUSIONS: Wheelchair rolling resistance is not always independent of velocity, and CR on linoleum can vary among wheelchairs by as much as seven-fold, and carpet can more than double CR. PMID- 14566158 TI - Sacral stress fracture in a female field hockey player. AB - We report a collegiate field hockey player who sustained a sacral fatigue-type stress fracture that manifested as persistent low back and leg pain. The diagnosis of sacral stress fracture was suggested by history and physical examination and confirmed by magnetic resonance imaging. Our patient experienced complete resolution of symptoms after a 3-mo interval of activity restriction. This article describes the first reported case of a sacral stress fracture in a field hockey player. PMID- 14566159 TI - Bone overgrowth in the adult traumatic amputee. AB - Bone overgrowth of the residual limb after an amputation is a well documented complication in the pediatric amputee population. Bone overgrowth can cause pain, problems with skin breakdown, and poor prosthetic fit. There have been few reports of bone overgrowth in the adult amputee. Two cases of traumatic transfemoral amputations after extensive tissue damage are presented. Both patients successfully completed an in-patient amputee rehabilitation program and achieved functional ambulation with their prostheses. However, each developed distal residual limb pain within a year after their amputations that significantly limited the amount of time they could wear their prostheses and the distance they could walk. Radiographs demonstrated additional bone growth from the residual femur into adjacent soft tissues in both patients. These case examples demonstrate that bone overgrowth should be considered in the differential diagnosis of residual limb pain in the adult amputee. PMID- 14566160 TI - Functional mobility measures in older adults after hip fracture. PMID- 14566161 TI - Caretaker bias in the study of young children. PMID- 14566162 TI - Aripiprazole and extrapyramidal symptoms. PMID- 14566163 TI - Cognitive-behavioral therapy for children with anxiety disorders in a clinical setting: no additional effect of a cognitive parent training. AB - OBJECTIVE: To evaluate a 12-week cognitive-behavioral treatment program for children with anxiety disorders and the additional value of a seven-session cognitive parent training program. METHOD: Seventy-nine children with an anxiety disorder (aged 7-18 years) were randomly assigned to a cognitive behavioral treatment condition or a wait-list control condition. Families in the active treatment condition were randomly assigned to an additional seven-session cognitive parent training program. Semistructured diagnostic interviews were conducted with parents and children separately, before and after treatment and at 3 months follow-up. Questionnaires included child self-reports on anxiety and depression and parent reports on child's anxiety and behavioral problems. RESULTS: Children with anxiety disorders showed more treatment gains from cognitive-behavioral therapy than from a wait-list control condition. These results were substantial and significant in parent measures and with regard to diagnostic status, but not in child self-reports. In the active treatment condition, children improved on self-reported anxiety and depression, as well as on parent reports on their child's anxiety problems. These results were equal for clinically referred and recruited children. Child self-reports decreased to the normal mean, whereas parents reported scores that were lower than before treatment but were still elevated from the normal means. No significant outcome differences were found between families with or without additional parent training. CONCLUSIONS: Children with anxiety disorders profited from cognitive behavioral therapy. Children improved equally whether or not additional parent training was offered. PMID- 14566164 TI - Neurochemical analyses in pediatric obsessive-compulsive disorder in patients treated with cognitive-behavioral therapy. AB - OBJECTIVE: To investigate neurochemical changes in the caudate nucleus of pediatric obsessive-compulsive disorder (OCD) patients before and after cognitive behavioral therapy (CBT), and to examine corresponding changes in symptom severity. METHOD: Single-voxel proton magnetic resonance spectroscopic (1H-MRS) examination of the left caudate was conducted in 21 treatment-naive children, aged 6 to 16 years, before and after 12 weeks of CBT. Subjects were measured at baseline and posttreatment by the Yale-Brown Obsessive Compulsive Scale for Children, Hamilton Depression Rating Scale, and Hamilton Anxiety Rating Scale. RESULTS: No significant changes in caudate neurochemistry were observed in OCD patients before and after CBT despite unambiguous improvement in OCD symptoms, depression, and anxiety. CONCLUSIONS: Findings suggest that reduction in caudate Glx may be specific to SSRI treatment and not due to a more generalized treatment response or spontaneous improvement of symptoms. Differential sets of pathophysiologic and treatment response markers may moderate/mediate the effects of particular treatments on outcome. PMID- 14566165 TI - Melatonin improves health status and sleep in children with idiopathic chronic sleep-onset insomnia: a randomized placebo-controlled trial. AB - OBJECTIVE: To investigate the effect of melatonin treatment on health status and sleep in children with idiopathic sleep-onset insomnia. METHOD: A randomized, double-blind, placebo-controlled trial was conducted in a Dutch sleep center, involving 62 children, 6 to 12 years of age, who suffered more than 1 year from idiopathic chronic sleep-onset insomnia. Patients received either 5 mg melatonin or placebo at 7 pm. The study consisted of a 1-week baseline period, followed by a 4-week treatment. Health status was measured with the RAND General Health Rating Index (RAND-GHRI) and Functional Status II (FS-II) questionnaires. Lights off time, sleep onset, and wake-up time were recorded in a diary, and endogenous dim light melatonin onset was measured in saliva. RESULTS: The total scores of the RAND-GHRI and FS-II improved significantly more during melatonin treatment compared to placebo. The magnitude of change was much higher in the melatonin group than in the placebo group, with standardized response means for the RAND GHRI of 0.69 versus 0.07 and for the FS-II of 1.61 versus 0.64. Melatonin treatment also significantly advanced sleep onset by 57 minutes, sleep offset by 9 minutes, and melatonin onset by 82 minutes, and decreased sleep latency by 17 minutes. Lights-off time and total sleep time did not change. CONCLUSIONS: Melatonin improves health status and advances the sleep-wake rhythm in children with idiopathic chronic sleep-onset insomnia. PMID- 14566166 TI - Major depressive disorder in adolescents exposed to a friend's suicide. AB - OBJECTIVE: To evaluate whether the risk of DSM-III major depressive disorder (MDD) is uniform across the 6 months after adolescent exposure to a friend's suicide, and to examine potential moderating or mediating processes that may influence the risk of new-onset MDD. METHOD: One-month incidence rates of MDD were compared between 129 adolescents who were exposed to a friend's suicide between December 1988 and March 1991 and 145 similar-aged, unexposed community controls participating in the Youth Exposed to Suicide study. Clinical, family, and social factors that antedated the exposure were examined as predictors of new onset MDD within 1 month of the suicide. RESULTS: Exposed adolescents had a markedly increased risk of developing new-onset MDD that was restricted to a narrow period of time, within 1 month after exposure. In contrast, there were no differences in the incidence of MDD between the groups in months 2 to 6. Past history of alcohol abuse increased the risk of exposure to suicide, which subsequently increased the risk of new-onset MDD within 1 month of exposure. Exposed youths who had both a family history of MDD and feelings of accountability for the death were at considerably increased risk of new-onset MDD. CONCLUSIONS: For adolescents exposed to a friend's suicide, events surrounding the death interact with family history of MDD to greatly increase the risk for new-onset MDD. PMID- 14566167 TI - Sexual abuse and suicidal behavior: a model constructed from a large community sample of adolescents. AB - OBJECTIVE: To investigate relationships between self-reported sexual abuse, depression, hopelessness, and suicidality in a community sample of adolescents. METHOD: In 1995, students (mean age 13 years) from 27 high schools in Australia (n = 2,603) completed a questionnaire including measures of depressive symptoms (Center for Epidemiologic Studies Depression Scale), hopelessness, sexual abuse, and suicidality. Data analysis included logistic regression. RESULTS: Sexual abuse is associated with suicidality, both directly and indirectly through hopelessness and depressive symptoms in the model developed. High suicide risk (behavior exceeding thoughts such as plans and threats, or deliberate self injury) is strongly correlated with suicide attempts (odds ratio 28.8, 95% CI 16 52, p <.001). Hopelessness is associated with high suicide risk only, whereas depressive symptoms are associated with high suicide risk and attempts. Hopelessness is more strongly associated with sexual abuse in boys than girls. Depressive symptoms are more strongly associated with high suicide risk in girls than boys. Controlling for other variables, sexual abuse is independently associated with suicide attempts in girls but not boys. CONCLUSIONS: Clinical assessment should consider gender differences in relationships between sexual abuse, depressive symptoms, hopelessness, and suicidality. Sexually abused girls may be at increased risk of attempting suicide, independent of other psychopathology. PMID- 14566168 TI - Salivary cortisol responses to dexamethasone in adolescents with posttraumatic stress disorder. AB - OBJECTIVE: Previous studies of adults with posttraumatic stress disorder (PTSD) have found various abnormalities in the regulation of the hypothalamic-pituitary adrenal axis, including enhanced suppression of cortisol following low-dose dexamethasone. The purpose of the present study was to investigate salivary cortisol responses to low-dose dexamethasone in adolescents with PTSD. METHOD: Forty-eight adolescents (20 with current PTSD, 9 trauma controls without PTSD, and 19 healthy nontraumatized controls) were enrolled in the study. On day 1, baseline saliva samples were obtained at 8 a.m. and 0.5 mg of dexamethasone was administered at 11 p.m. Cortisol and dexamethasone levels were assessed at 8 a.m. the following day. RESULTS: Adolescents with current PTSD showed no difference in the suppression of salivary cortisol in response to low-dose (0.5 mg) dexamethasone compared to trauma controls without PTSD and nontraumatized controls. More severely affected PTSD subjects with co-occurring major depression showed higher pre- and post-dexamethasone salivary cortisol levels compared to controls. CONCLUSIONS: The present study did not find evidence for enhanced suppression of salivary cortisol at 8 a.m. following low-dose dexamethasone in multiply traumatized adolescents with PTSD. This result differs from findings in adults with PTSD. Further investigations of hypothalamic-pituitary-adrenal axis abnormalities in traumatized children and adolescents are needed. PMID- 14566169 TI - Detection and outcomes of substance use disorders in adolescents seeking mental health treatment. AB - OBJECTIVE: To compare detection rates, service use, and outcomes of substance use disorder (SUD) in adolescents seeking mental health treatment. METHOD: Adolescents (n = 237) and their parents or caregivers completed parallel, self administered versions of the Adolescent Treatment Outcomes Module (ATOM) at intake and 6-month follow-up. SUD was assessed using the Diagnostic Interview Schedule for Children (DISC). RESULTS: Although 42 (16.6%) adolescents seeking mental health treatment met DISC criteria for any SUD at baseline, clinicians detected only 19 of these (kappa = 0.51). Sensitivity and specificity were high for a screening tool for adolescent SUD (95.2 and 82.0, respectively). Adolescents with undetected SUD were less likely to receive SUD services and to have more legal problems at 6-month follow-up compared to adolescents with detected SUD. CONCLUSIONS: SUD rates may be high in adolescents seeking mental health treatment, confirming the need for routine screening in this population. Results confirm that a large gap exists between the need for and access to SUD treatments, which may contribute to poorer outcomes for all adolescents with SUD. PMID- 14566170 TI - Reported maltreatment among clinic-referred children: implications for presenting problems, treatment attrition, and long-term outcomes. AB - OBJECTIVE: To examine the treatment implications of a reported history of maltreatment in a sample of 343 children referred to Los Angeles area mental health clinics for emotional and behavioral problems. METHOD: Child Protective Service records identified 161 of the 343 families as having a documented history of maltreatment. Parent reports of child behavior problems were obtained following clinic intake and at 2-year follow-up, and attrition was assessed via medical records. The authors examined the associations between a history of maltreatment and severity of child presenting problems, treatment attrition, and long-term mental health outcomes. RESULTS: Compared to other clinic-referred children, youngsters with a history of maltreatment entered treatment with more externalizing behavior problems, were more likely to exit treatment early and without therapist consent, and continued to show greater externalizing problems 2 years later. CONCLUSIONS: Maltreated children have significant mental health needs that may not be well addressed currently in community mental health clinics. PMID- 14566171 TI - Do executive deficits and delay aversion make independent contributions to preschool attention-deficit/hyperactivity disorder symptoms? AB - OBJECTIVE: To test whether deficits in executive function and delay aversion make independent contributions to levels of attention-deficit/hyperactivity disorder (ADHD) symptoms exhibited by preschool children. METHOD: One hundred fifty-six children between 3 and 5.5 years old (78 girls and 78 boys) selected from the community completed an age-appropriate battery of tests measuring working memory, set shifting, planning, delay of gratification, and preference for delayed rewards. Parents completed a clinical interview about their children's ADHD symptoms. RESULTS: Analysis of test performance revealed two factors: executive dysfunction and delay aversion. Multivariate analysis demonstrated that when other factors (i.e., age, IQ, and conduct problems) were controlled, executive dysfunction and delay aversion each made significant independent contributions to predictions of ADHD symptoms. CONCLUSIONS: Preschool ADHD symptoms are psychologically heterogeneous. Executive dysfunction and delay aversion may represent two distinct and early appearing neurodevelopmental bases for ADHD symptoms. PMID- 14566172 TI - Three informants' reports of child behavior: parents, teachers, and foster parents. AB - OBJECTIVE: To evaluate whether disagreements in reporting child behavior problems between biological parents and foster parents and teachers are related to depressive symptoms in the biological parent. METHOD: Child Behavior Checklists and Teacher's Report Forms were completed by parents, foster parents, and teachers on 95 foster children between ages 5 and 16 during 1990-1991. Depressive symptoms in biological parents were assessed using the Center for Epidemiologic Studies Depression Scale. Structural equation modeling was used to estimate the relation between parental depressive symptoms and the discrepancies between their reports and those of two other informants. RESULTS: In both the internalizing and externalizing behavior models, a significant relation was found between parents' depressive symptoms and their discrepancies in reporting, but not with child behavior. This relation was stronger for internalizing than externalizing models. CONCLUSIONS: Using foster parents (and teachers) as informants offered a unique opportunity to view the reports of biological parents in comparison to two independent reports. Foster parents have a similar observational situation as biological parents but bring a less emotionally invested perspective to behavior observation, even when they are relatives. It is likely that depressive symptoms in parents distort their reports of their children's behavior. Clinicians would be wise to supplement parental with other informant reports. PMID- 14566173 TI - A study of parent ratings of internalizing and externalizing problem behavior in 12-year-old twins. AB - OBJECTIVE: Studies on 3-, 7-, and 10-year-old twins' internalizing and externalizing problems have emphasized the importance of understanding sources of agreement and disagreement between maternal and paternal ratings. A psychometric model that assumes that each parent assesses rater-specific aspects of the child's behavior provided the best explanation for parental disagreement. This study investigates two models that have been used to explain the agreement and disagreement between mothers and fathers in the ratings of their children. METHOD: Child Behavior Checklists filled in by mothers and fathers were collected for a sample of 1,481 twelve-year-old twin pairs. Genetic and environmental influences on internalizing and externalizing problems were estimated using models that corrected for rater bias, rater-specific effects, and unreliability. RESULTS: The psychometric model fitted the data significantly better than a rater bias model. Significant influences of genetic, shared, and nonshared environmental factors were found for internalizing and externalizing behavior. Parent-specific views, rater bias, and unreliability were significant. CONCLUSIONS: The best-fitting model implies that disagreement between parents is due to the fact that mothers and fathers provide information from their own perspective. This information should be seen as important and adding to the diagnostic formulation rather than as a point of disagreement. The finding that internalizing and externalizing problems are influenced by genetic and environmental factors fosters the understanding that it is the interaction of nature and nurture that puts children at risk for common behavioral disorders. PMID- 14566174 TI - Persistence of psychiatric disorders in pediatric settings. AB - OBJECTIVE: To examine the longitudinal course of subthreshold and full-criteria child psychiatric disorders. METHOD: Nine hundred ninety-six children, aged 4 to 9 years, from a representative sample of pediatric primary care practices participated in a 1-year follow-up in 1989-1990. Parent interviews included the Diagnostic Interview Schedule for Children, measures of psychosocial problems, stress, social support, possible child abuse, and sociodemographics. RESULTS: The initial prevalence of full and subthreshold disorders was 18.8% and 14.0%, respectively. Full and subthreshold disorders were associated with impairment. Approximately 37% of children with full disorders had persistent disorders. Persistence was more common among boys than girls, particularly when comorbidity was present. Children with initial subthreshold disorders only were five times more likely than those without disorder to develop a full disorder. Persistence of full disorders was associated with high initial symptoms. Among boys, persistence was linked to possible child abuse, whereas among girls it was associated with full externalizing disorders that were accompanied by subthreshold internalizing disorders. CONCLUSIONS: Intervention may be particularly crucial when a high number of symptoms and/or both internalizing and externalizing problems are present. Underscoring the importance of family centered rather than child-centered interventions, boys whose parents reported possible child abuse were more likely to have persistent disorders. PMID- 14566175 TI - Psychological morbidity and caregiver burden in parents of children with Tourette's disorder and psychiatric comorbidity. AB - OBJECTIVE: To investigate the mental health and caregiver burden in parents of children with Tourette's disorder (TD) compared with parents of children with asthma. METHOD: A cross-sectional cohort survey was conducted at TD and pediatric asthma hospital outpatient clinics over a 6-month period. Main outcome measures were parent mental health (General Health Questionnaire [GHQ]-28) and caregiver burden (Child and Adolescent Impact Assessment) scores. RESULTS: The response rate achieved was 89.7%. Of the parents of children with TD, 76.9% achieved caseness on the GHQ-28 compared with 34.6% of the parents of children with asthma; this effect remained significant after controlling for demographic variables. Parents of children with TD also experienced greater caregiver burden, and this burden was significantly correlated with GHQ caseness. CONCLUSIONS: Parents of children with TD are at risk of psychiatric morbidity; an intervention targeting caregiver burden might be helpful in reducing this. PMID- 14566176 TI - Meta-analysis: formulation and interpretation. PMID- 14566177 TI - Potential of magnetic resonance lymphography with intrapulmonary injection of gadopentetate dimeglumine for visualization of the pulmonary lymphatic basin in dogs: preliminary results. AB - PURPOSE: To evaluate a new approach of magnetic resonance (MR) lymphography with intraalveolar injection of a conventional extracellular contrast agent (gadopentetate dimeglumine) for imaging lymphatic basin draining from specific portions of the lung. METHODS: Three-dimensional T1-weighted spoiled gradient recalled echo MR sequence images were acquired serially before and for 40 minutes after intraalveolar injection of gadopentetate dimeglumine in a total of 14 anesthetized beagle dogs. Six of these dogs received 1 mL undiluted and low concentration (75%) contrast agent into the same portion of the right caudal lobe during a 7-day interval. In all dogs, including these 6 dogs, MR lymphography was repeated with injection of the low-concentration contrast agent into different lung regions at 7-day intervals to evaluate the differences of the visualized draining lymphatic station. Lymphatic enhancement was quantified by percent increases of signal intensity against precontrast. Postmortem examination of the lymphatic anatomy was performed in 7 of these animals. RESULTS: In all dogs, the lymphatic station draining from the injection sites was visualized within 5 minutes after contrast injection. The maximum percent increase of signal intensity of the same middle tracheobronchial lymph nodes was significantly greater with a low-concentration (75%) contrast agent than with an undiluted one in the same 6 dogs (n = 6, 247.6 +/- 30.5% vs. 204.2 +/- 33.8%; P < 0.01). Different lymphatic stations draining from the different injection sites were visualized in all dogs. In a total of 12 MR studies that showed extended nodal enhancement after injection of the low-concentration contrast agent, the enhancement peak of the most proximal nodes (n = 12) from the injection sites appeared earlier than that of their distant nodes (n = 12), with a maximum percent increase of signal intensity of 249.8 +/- 42.4%. The visualized lymph nodes were found in the appropriate locations postmortem, with significant correlation for nodal sizes (r = 0.965; P < 0.0001). CONCLUSION: MR lymphography with low-concentration gadopentetate dimeglumine can quickly and sufficiently visualize the drainage lymphatic station from specific lung portions, and may have the potential of sentinel node mapping in lung cancer. PMID- 14566178 TI - Optimal contrast application for cardiac 4-detector-row computed tomography. AB - RATIONALE AND OBJECTIVES: This study was designed to determine the optimal contrast protocol for 4-detector-row computed tomography angiography of the heart. METHODS: Sixty patients were randomly assigned to 1 of 4 groups with 300 and 400 mg/mL iodine concentrations and 2.5 and 3.5 mL/s flow rates. Contrast density was measured in the left ventricular cavity and coronary arteries. RESULTS: Low iodine concentration injected at slow flow rate (0.75 g iodine/s) resulted in acceptable contrast enhancement in only 53.8% of the patients. There was no significant difference between low contrast concentration injected at high flow rate and high contrast concentration injected at slow flow rate ( approximately 1 g iodine/s). High contrast concentration administered with high flow rates (1.4 g iodine/s) may result in an enhancement above 350 Hounsfield units (HU) and interfere with coronary calcifications. CONCLUSIONS: The injection of approximately 1 g iodine/s resulted in an optimal (250-300 HU) contrast enhancement for cardiac 4-detector-row computed tomography. PMID- 14566179 TI - Imaging findings after fat graft interposition in an injured growth plate: an experimental study in rabbits. AB - RATIONALE AND OBJECTIVES: To assess the magnetic resonance (MR) features of fat graft interposition in experimentally induced growth plate injury, to correlate these features with histologic findings, and to compare the growth disturbances of fat-grafted and nonfat-grafted growth plate injuries. METHODS: The growth plate injury model was created in 20 skeletally immature rabbits. In 14 rabbits, a bilateral growth plate defect was made in the medial half of the proximal tibia. The defect of the right tibia was filled with autologous fat; the left tibia was left empty. In another 6 rabbits, a unilateral growth plate defect was created with (n = 3) or without (n = 3) fat graft interposition in the right tibia. MR imaging was performed at 2, 4, and 12 weeks postoperatively using a 1.5 T MR unit. T1-weighted images, T2-weighted images, gradient echo images, and contrast-enhanced T1-weighted images were obtained in the coronal plane. The authors evaluated the MR features of the growth plate defects with or without fat graft, and the remaining growth plate. The length and angulation of the grafted tibia were compared with those of the nonfat-grafted tibia. RESULTS At 2 weeks postoperatively, the grafted area showed low signal intensity mixed with high signal intensity (71%) on T1-weighted imaging. At 2 weeks, most of the growth plate defect in the control animals showed low signal intensity in all image sequences. At 4 weeks, most of the grafted area showed low signal intensity on T2 weighted imaging. A bone bridge was revealed in 15% of the grafted tibia and in 62% of the control animals at 4 weeks. On gradient echo images, high signal intensity of the remaining growth plate was preserved in grafted tibias, but in the control animals this became invisible 4 weeks postoperatively. At 12 weeks, most of growth plate defect area became isointense on all image sequences in grafted tibia and in the control animals. Histologically, the fat-grafted area was replaced by fibrous connective tissues with thin, trabecular bone formation. Growth plate defects of the control animals were filled with mature fatty marrow with trabecular bone formation. The varus angle and medial length of the tibia were significantly different between fat-grafted tibias at 4 weeks and 12 weeks postoperatively. CONCLUSION: MR imaging was helpful for evaluating growth plate modifications and subsequent changes after fat graft interposition. Fat graft interposition had a role in reducing growth disturbance in growth plate injury. PMID- 14566180 TI - A new animal model for testing renal tolerance of contrast agents. AB - RATIONALE AND OBJECTIVES: To develop a new animal model for the evaluation of the renal tolerance of contrast agent and its application to diatrizoate and iotrolan injections in comparison to saline and mannitol solution. MATERIALS AND METHODS: The nephrotoxic potential of intravenously injected contrast agents in Beagle dogs was determined from the effects on the pharmacokinetics of creatinine injected immediately after these agents in comparison with the kinetics of creatinine injected alone. The kinetic parameters evaluated included plasma half life, volume of distribution, and clearance. In a series of studies, the experimental conditions were optimized in terms of creatinine dose and the effects of diatrizoate, iotrolan, saline, and mannitol solution on the pharmacokinetic parameters of creatinine were determined. RESULTS: Bolus injection of creatinine into dogs followed a dose-linear pharmacokinetic behavior with a terminal half-life of 90 minutes (10 mg/kg) and 108 minutes (20 mg/kg), respectively, and a clearance of 5 mL/min/kg. Injection of diatrizoate decreased the clearance and the half-life of creatinine significantly. A mannitol solution with the same osmolality as diatrizoate showed the same effect whereas physiological saline did not affect pharmacokinetic parameters of creatinine. The isotonic iotrolan was intermediate between saline and diatrizoate in its effects on creatinine clearance. CONCLUSION: The effect of contrast agents on pharmacokinetic parameters of creatinine injected immediately after the administration of the agents in dogs might be a useful model for the evaluation of renal tolerance. In a cross-over study, iotrolan was not statistically separable from diatrizoate. PMID- 14566181 TI - Comparison of arterial spin-labeling techniques and dynamic susceptibility weighted contrast-enhanced MRI in perfusion imaging of normal brain tissue. AB - OBJECTIVES: To evaluate relative cerebral blood flow (rCBF) in normal brain tissue using arterial spin-labeling (ASL) methods and first-pass dynamic susceptibility-weighted contrast-enhanced (DSC) magnetic resonance imaging (MRI). METHODS: Sixty-two patients with brain metastases were examined on a 1.5 T-system up to 6 times during routine follow-up after stereotactic radiosurgery. Perfusion values in normal gray and white matter were measured using the ASL techniques ITS FAIR in 38 patients, Q2TIPS in 62 patients, and the first-pass DSC echo-planar (EPI) MRI after bolus administration of gadopentetate dimeglumine in 42 patients. Precision of the ASL sequences was tested in follow-up examinations in 10 healthy volunteers. RESULTS: Perfusion values in normal brain tissue obtained by all sequences correlated well by calculating Pearson's correlation coefficients (P < 0.0001) and remained unchanged after stereotactic radiosurgery as shown by analysis of variance (P > 0.05). CONCLUSION: Both ASL and DSC EPI MRI yield highly comparable perfusion values in normal brain tissue. PMID- 14566182 TI - Comparison of the detectability of high- and low-contrast details on a TFT screen and a CRT screen designed for radiologic diagnosis. AB - RATIONALE AND OBJECTIVES: To evaluate the detection rate of fine details of a new thin-film transistor (TFT) grayscale monitor designed for radiologic diagnosis, compared with a type of cathode ray tube (CRT) screen used routinely for diagnostic radiology. METHODS: Fifteen radiographs of a statistical phantom presenting low- and high-contrast details were obtained and read out with an Agfa ADC compact storage phosphor system. Each radiograph presented 60 high-density (high-contrast) and 60 low-density (low-contrast) test bodies. Approximately half the test bodies contained holes with different diameters. Observers were asked to detect the presence or absence of a hole in the test body on a 5-point confidence range. The total of 1800 test bodies was reviewed by 5 radiologists on the TFT monitor (20.8 inches; 1536 x 2048 pixels; maximum luminance, 650 cd/m2; contrast, 600:1) and the CRT monitor (21 inches; P45 Phosphor; 2048 x 2560 pixels operated at 1728 x 2304 pixels; maximum luminance, 600 cd/m2; contrast, 300:1). The data were analyzed by receiver-operator characteristic analysis. RESULTS: For high contrast details, the mean area under the curve rated 0.9336 for the TFT monitor and 0.9312 for the CRT monitor. For low-contrast details, the mean area under the curve rated 0.9189 for the TFT monitor and 0.9224 for the CRT monitor. At P .05. Image quality of the FFS anatomic structures were, thus, not statistically different from the HHS. Although the color cosmetic was different for the HHS directional (nonvelocity) map, only 9% of 150 total findings (including structural abnormalities and flow features, none of which were critical) were missed, whereas the other 91% regurgitant, shunt, stenosis flow features or heart structure were imaged adequately by the HHS in this population. CONCLUSIONS: Implementing high-frequency transducers and programs optimized for tissue and flow imaging on the HHS should provide images of sufficient quality for targeted echocardiography examinations to determine the presence, absence, or status of congenital heart disease in newborns and young children. PMID- 14566292 TI - Time requirements of the standard echocardiogram: implications regarding limited studies. AB - BACKGROUND: Although no formal detailed definitions exist, a standard echocardiogram should easily be differentiated from a limited examination by its comprehensive nature and time requirement. OBJECTIVE: We evaluated the time required to perform each specific component of a comprehensive echocardiography examination and applied these data to compare the time required to complete 3 limited imaging protocols. METHODS: In all, 7 experienced sonographers were timed in their performance of a comprehensive echocardiographic examination on a healthy patient. A total of 3 limited imaging protocols were then constructed to evaluate for pericardial effusion, wall motion, and left ventricle size. The time necessary to complete each protocol was calculated by adding the average time required to perform all of its components and was compared with the comprehensive examination. RESULTS: The imaging time required to complete a standard study was 26 +/- 5.4 minutes (range: 20-32). The percentage of total study time spent on 2 dimensional imaging, M-mode, color Doppler, and spectral components was 31 +/- 2.9%, 11 +/- 3.6%, 17 +/- 2.9%, and 39 +/- 5.0%, respectively. The calculated time needed to perform a limited protocol was less than 5 minutes, resulting in a greater than 80% reduction in time compared with the standard examination. CONCLUSION: Time analysis of components of the standard echocardiography examination can be used to construct appropriate, time-efficient, limited, cardiac ultrasound imaging examinations and differentiate limited from comprehensive studies. PMID- 14566293 TI - Myocardial global performance index as a predictor of in-hospital cardiac events in patients with first myocardial infarction. AB - OBJECTIVE: We sought to assess the ability of a Doppler index of global myocardial performance (MPI), measured at entry, to predict inhospital cardiac events in a series of patients with first acute myocardial infarction (AMI). METHODS: A complete 2-dimensional and Doppler echocardiographic examination was performed within 24 hours of arrival at the coronary care department in 96 patients (81 men and 15 women; mean age 58 +/- 9 years) with first AMI. Patients were divided a posteriori into 2 groups according to their inhospital course: group 1 comprised 75 patients with an uneventful course and group 2 comprised 21 patients with a complicated inhospital course (death, heart failure, arrhythmias, or post-AMI angina). RESULTS: There were no significant differences between the 2 groups with regard to history of hypertension, diabetes mellitus, hypercholesterolemia, site and size of infarction, and conventional parameters of diastolic function. However, patients with complications were significantly older (63 +/- 10 vs 55 +/- 8 years, P =.005) and had higher wall-motion score index and left ventricular end-systolic volume compared with patients without events (1.84 +/- 0.27 vs 1.52 +/- 0.30, P =.001; and 66 +/- 29 vs 47 +/- 21 mL, P =.009, respectively), whereas the ejection fraction was reduced (40 +/- 10% vs 52 +/- 10%, P =.0001). The mean value of the MPI was significantly higher in patients with cardiac events than in those without events (0.65 +/- 0.20 vs 0.43 +/- 0.16, P =.0001). A MPI >/= 0.47 showed a sensitivity of 90% and specificity of 68% for identifying patients with events, on the basis of the receiver operator curve. In a multivariable model, the MPI at admission remained independently predictive of inhospital cardiac events (odds ratio 15.6, 95% confidence interval 2.4-99, P =.003). CONCLUSION: These data suggest that in the acute phase of AMI, the MPI measured at entry may be useful to predict which patients are at high risk for inhospital cardiac events. PMID- 14566294 TI - Assessment of left ventricular systolic and diastolic function by Doppler tissue imaging in patients with preinfarction angina. AB - BACKGROUND: The aim of this study was assessment of left ventricular (LV) systolic and diastolic function by pulsed wave Doppler tissue imaging (DTI) in patients with or without preinfarction angina in acute myocardial infarction. METHODS: We prospectively evaluated 31 consecutive patients (4 women, 27 men; age 58 +/- 10 years) with a first acute myocardial infarction. LV systolic and diastolic function was assessed by classic methods and DTI on the third day during acute myocardial infarction. Patients were divided into 2 groups according to the presence (group 1; n = 10) or absence (group 2; n = 21) of preinfarction angina. Mitral inflow velocities and early diastolic mitral annular velocity (Em), late diastolic mitral annular velocity (Am), peak systolic mitral annular velocity, Em/Am, the ratio of early diastolic mitral inflow velocity (E) to Em, and myocardial performance index were calculated by DTI. RESULTS: Group 1 had significantly higher Em and Em/Am than group 2 (11.3 +/- 3.34 cm/s vs 7.4 +/- 2.07 cm/s, P <.0001; 1.01 +/- 0.38 cm/s vs 0.6 +/- 0.2 cm/s, P =.001, respectively). The E/Em ratio and myocardial performance index were significantly lower in group 1 than in group 2 (5.1 +/- 2.92 vs 8.10 +/- 3.15, P=.018; 0.49 +/- 0.15 vs 0.65 +/- 0.24, P =.042, respectively). Wall-motion score index was lower in those with preinfarction angina than in those without (1.6 +/- 0.36 vs 1.9 +/- 0.39; P =.04, respectively). Peak systolic mitral annular velocity and Am were not statistically different between groups (9.4 +/- 1.84 vs 8.3 +/- 2.03, P =.172; 11.7 +/- 3.07 vs 12.1 +/- 3.34, P =.72, respectively). There were no significant differences between the 2 groups regarding transmitral E velocity, atrial contraction mitral inflow velocity (A), E/A ratio, isovolumetric relaxation time, and deceleration time of the mitral E wave (P =.91, P =.08, P =.58, P =.81, and P =.71, respectively). CONCLUSION: LV diastolic function was better in patients with preinfarction angina than in patients without. This condition could not be detected by conventional mitral inflow Doppler velocities, but could be detected by DTI. This preliminary evidence shows that DTI is better than conventional mitral Doppler indices in the assessment of a favorable LV diastolic function in patients with preinfarction angina. PMID- 14566295 TI - Doppler tissue analysis of mitral annular velocities: evidence for systolic abnormalities in patients with diastolic heart failure. AB - BACKGROUND: The presence of signs and symptoms of heart failure (HF), abnormal diastolic function and an ejection fraction > 45%, have been defined as diastolic HF (DHF). However, a cut-off value of 45% for ejection fraction seems arbitrary as mild systolic dysfunction may be overlooked. It was the goal of this study to assess the additive information derived from Doppler tissue imaging for patients with DHF. METHODS: As a measure of left ventricular (LV) long-axis function, systolic and diastolic velocities of the mitral annulus (peak, peak early, and peak late) derived from pulsed Doppler tissue imaging were assessed in 36 asymptomatic control subjects, 36 patients with DHF, and 35 patients with systolic HF (SHF). As a measure of overall LV performance, the Tei index (isovolumic contraction time and isovolumic relaxation time divided by ejection time) was assessed. RESULTS: In the DHF group, peak systolic annular velocity was reduced (7.1 +/- 1.2 cm/s) as compared with the control group (9.0 +/- 1.2 cm/s, P <.05), and was even lower in the SHF group (5.0 +/- 0.7 cm/s, P <.01 SHF group vs DHF/control groups). The Tei index was increased in the DHF group (0.53 +/- 0.14) in comparison with the control group (0.39 +/- 0.07, P <.05), and was highest in the SHF group (0.94 +/- 0.43, P <.01 SHF group vs control/DHF groups). Using peak systolic annular velocity < 7.95 cm/s as a cut-off value (derived from receiver operating characteristic curve analysis), patients with DHF were separated from control subjects with a sensitivity of 83% and a specificity of 83%. A Tei index > 0.43 separated patients with DHF and control subjects with a sensitivity of 79% and a specificity of 72%. CONCLUSION: Systolic long-axis LV function is also impaired in patients with DHF, resulting in feasible diagnosis of DHF by Doppler tissue imaging analysis of LV long-axis function and overall LV function with the Tei index. PMID- 14566296 TI - Myocardial contrast echocardiography (MCE) with triggered ultrasound does not cause premature ventricular complexes: evidence from PB127 MCE studies. AB - Previous studies suggest that myocardial contrast echocardiography using high mechanical index triggered ultrasound can be associated with increased frequency of the premature ventricular complex (PVC). However, this association has not been systematically examined. PB127 (Point Biomedical Corp, San Carlos, Calif) is a novel microsphere designed for evaluation of myocardial perfusion with ultrasound. PB127 myocardial contrast echocardiography was performed with triggered harmonic power Doppler in early/mid diastole (mechanical index .999) and was lower than untriggered intervals (P =.001) in B, suggesting that triggers do not cause PVC. PB127 does not cause increase PVC frequency during or after imaging with triggered ultrasound at mechanical index of 1. PMID- 14566297 TI - Increase in atrial size in long-term survivors of heart transplant. AB - Atrial size in recipients of orthotopic heart transplant (OHTX) who have long term survival is not well described in the literature. We reviewed 2-dimensional echocardiograms of 14 recipients of OHTX who survived at least 5 years. Apical 4 chamber images were used for measurements starting at 1-year post-OHTX. The recipients of OHTX were 8 (57%) men and 6 (43%) women with a mean age of 43.6 +/- 12.1 (20-60) years and mean survival of 9.5 +/- 2.6 (5.8-14.4) years. All chambers increased with time post-OHTX (r > 0.83, P <.002). The changes in left atrium (LA) and right atrium (RA), remnants of donor LA and RA, and remnants of recipients' LA areas correlated inversely with patient survival post-OHTX (r > 0.83, P <.002). The change in RA recipient remnant area did not correlate with survival (r = 0.58, P =.06). In recipients of OHTX with long-term survival, there is a significant correlation between the echocardiographic size of LA and RA, and donor and recipient remnants with time and survival. PMID- 14566298 TI - Association of newer diastolic function parameters with age in healthy subjects: a population-based study. AB - OBJECTIVES: We sought to determine if newer Doppler diastolic function parameters are associated with age and to define age-stratified reference ranges for these parameters in a population-based setting. BACKGROUND: Although newer Doppler parameters aid in the evaluation of diastolic function and filling pressures, age specific reference ranges are poorly described. METHODS: Randomly selected residents of Olmsted County, Minn, age >/= 45 years were studied (n = 2042) prospectively. Those without a history of cardiovascular disease or abnormal 2 dimensional echocardiograms were selected. Measured parameters included the transmitral early (E) and late (A) filling velocities at rest and with Valsalva, A duration, pulmonary venous atrial reversal duration, lateral and septal early diastolic mitral annular velocities, E/lateral early diastolic mitral annular velocity, E/septal early diastolic mitral annular velocity, and the index of left ventricular myocardial performance. RESULTS: A total of 1012 subjects met the inclusion criteria. Both A duration (r = 0.18, P <.001) and pulmonary venous atrial reversal duration (r = 0.28, P <.001) were longer in older subjects, whereas their difference showed a marginal correlation with age (r = 0.07, P =.036). The E/A ratio at peak Valsalva correlated with age (r = -0.54, P <.001) whereas the change in E/A with Valsalva (corrected for E-A fusion) showed a marginal correlation with age (r = 0.08, P =.032). Both E/lateral early diastolic mitral annular velocity (r = 0.39, P <.001) and E/septal early diastolic mitral annular velocity (r = 0.27, P <.001) increased with age. Left ventricular myocardial performance showed a marginal correlation with age (r = 0.08, P =.014). Age specific reference ranges (median, 5th and 95th percentiles) for all parameters are provided. CONCLUSIONS: All newer Doppler diastolic function parameters showed a variable but significant correlation with age. Age-specific reference values will aid in the interpretation of diastolic function parameters. PMID- 14566299 TI - Pressure half-time predicts hemodynamically significant pulmonary regurgitation in adult patients with repaired tetralogy of fallot. AB - Pulmonary regurgitation (PR) is associated with adverse outcomes late after tetralogy of Fallot repair. Accurate assessment of PR in these patients is, therefore, fundamental to their clinical treatment; however, accurate ultrasound markers of severity are as yet poorly defined. This is a prospective study of 34 adult patients with repaired tetralogy of Fallot. Cardiac magnetic resonance imaging was used to assess the PR fraction and its hemodynamic significance on the right ventricular volumes. Regurgitant fractions >/= 20% were associated with significant increases in right ventricular end-diastolic volumes. Echocardiographic continuous wave Doppler profiles of the PR jet were used to calculate pressure half-time. Pulmonary pressure half-time < 100 milliseconds was found to be a good indicator of hemodynamically significant regurgitation. This measure is highly reproducible and easily accessible. PMID- 14566300 TI - Echocardiographic predictors of failure in patients undergoing coil occlusion of patent ductus arteriosus. AB - In this study, we sought to determine whether the risk of failure of coil occlusion of patent ductus arteriosus (PDA) could be predicted on the basis of echocardiographic variables. The echocardiographic characteristics of patients in whom PDA coil occlusion failed were compared with those in whom the procedure was successful. A total of 5 variables were evaluated: PDA diameter (PDAd); indexed PDAd; PDA shape; left ventricular end-diastolic diameter; and the presence of flow reversal in the descending aorta. We found that 2 variables related to the size of the duct (PDAd and PDAd/body surface area), and 2 related to the magnitude of the shunt (left ventricular end-diastolic diameter and flow reversal) were positively associated with failure (P <.05). PDAd and flow reversal had the greater effect in each group and remained significant when they were put into a logistic regression model to predict failure (P =.004 and.053, respectively). In conclusion, echocardiographic variables can predict risk of failure in patients undergoing PDA coil occlusion. PMID- 14566301 TI - Is cardiac catheterization a prerequisite in all patients undergoing bidirectional cavopulmonary anastomosis? AB - Traditionally, all patients undergo cardiac catheterization before bidirectional cavopulmonary anastomosis (BCPA). The purpose of this study was to determine if preoperative catheterization is necessary when echocardiographic parameters appear favorable. A retrospective review was performed of all patients who underwent BCPA (n = 142) between February 1996 and May 2001. Echocardiographic criteria defining a favorable BCPA candidate included good ventricular function, moderate or less atrioventricular and semilunar valve regurgitation, absence of ventricular outflow tract obstruction, normal proximal branch pulmonary artery (PA) size, and low PA pressures estimated by PA band gradient or systemic PA shunt velocity. The median age at operation was 7 months (range: 2-11) and weight was 6.2 kg (range: 2.7-7.1). There were 73 unfavorable candidates. Patients with hypoplastic left heart syndrome (n = 23) and pulmonary atresia with intact septum (n = 15) predominated among the unfavorable group. All patients were catheterized. This provided additional information on PA pressures in 3 patients in the favorable group but did not defer operation or influence outcome (no mortality, prolonged pleural drainage, or longer intensive care department stay). All 3 patients are alive at a mean follow-up of 51 months. The 30-day mortality was 2% (4 of 151 patients), all in the unfavorable group. Overall, 20 patients (13%) required arterioplasty of PAs at the time of BCPA. BCPA can be performed with a low risk of morbidity and mortality in a wide range of patients. By using commonly acquired echocardiographic parameters, a low-risk subgroup of patients can be identified who can safely avoid preoperative cardiac catheterization. PMID- 14566302 TI - Novel uses of intracardiac echocardiography with a phased-array imaging catheter. AB - A newer phased-array ultrasound imaging catheter (AcuNav, Siemens, Moutainview, Calif) provides comprehensive anatomic and physiologic data during cardiac interventions. The role of this catheter in percutaneous closure procedures, transseptal ablative procedures, and valvular interventions has been reported. We describe an expanded role of intracardiac echocardiography using AcuNav imaging catheter (Siemens) in 2 clinical situations. PMID- 14566303 TI - The a-dip of aortic regurgitation. AB - Echocardiography has become the diagnostic technique of choice for delineating the intracardiac hemodynamics in a host of pathophysiologic states. Pressures and flows can be estimated or measured with enough accuracy to allow for clinical decision-making. We present a case with an unusual Doppler echocardiographic finding and discuss its derivation. PMID- 14566304 TI - The enigma of occult mitral regurgitation in a patient with cardiogenic shock. AB - In patients with mitral regurgitation (MR), there is a high pressure gradient between the left ventricle (LV) and left atrium that Doppler echocardiography can easily detect. We present a case where transthoracic echocardiography (TTE) failed to provide an accurate evaluation of a patient who presented in cardiogenic shock. A transesophageal echocardiogram diagnosed MR because of a flail leaflet caused by a ruptured papillary muscle in the setting of normal ventricular function. PMID- 14566305 TI - Chronic mitral regurgitation secondary to a perivalvular leak from remote trauma. AB - We report a rare case of mitral regurgitation as a result of perivalvular leak in a 49-year-old man with a history of blunt trauma. He presented with a 2-month history of progressive exertional dyspnea, angina, and heart failure. Preoperative transesophageal echocardiography demonstrated severe mitral regurgitation as a result of a perivalvular leak of the mitral valve that was not evident on transthoracic echocardiography and cardiac catheterization. PMID- 14566306 TI - Delayed presentation of left ventricular outflow tract aneurysm after penetrating cardiac trauma. AB - We report a case of posttraumatic left ventricular outflow tract aneurysm in a patient who had a stab injury to the chest requiring emergency operation 40 years previously. After apparent decades without symptoms, the patient presented with exertional dyspnea. Clinical and echocardiographic assessment revealed aortic regurgitation and left ventricular outflow tract aneurysm. Injuries to the chest wall that penetrate the heart and great vessels are life-threatening and require emergency operative intervention. However, these injuries rarely, as in this case, result in chronic cardiac aneurysm and aortic valvular incompetence. PMID- 14566307 TI - Restrictive left ventricular filling in hypereosinophilic syndrome as a result of partial cavity obliteration by an apical mass: a strain/strain rate study. AB - We present a case of hypereosinophilic syndrome with a large apical mass, which markedly reduced end-systolic left ventricular cavity volume. Standard blood pool Doppler findings showed a restrictive filling pattern. However, ultrasound derived regional strain rate and strain parameters suggested that wall deformation was not compromised, inferring that the restrictive filling pattern in this case was solely as a result of the reduction in end-systolic cavity size. PMID- 14566308 TI - ACC/AHA/ASE 2003 Guideline Update for the Clinical Application of Echocardiography: summary article. A report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines (ACC/AHA/ASE Committee to Update the 1997 Guidelines for the Clinical Application of Echocardiography). PMID- 14566309 TI - The fall and rise of chloramphenicol. PMID- 14566310 TI - The development of pediatric ophthalmology and strabismus in Egypt. PMID- 14566311 TI - Pediatric ophthalmology in Australia. PMID- 14566312 TI - Preschool vision screening: what should we be detecting and how should we report it? Uniform guidelines for reporting results of preschool vision screening studies. PMID- 14566313 TI - Efficacy and safety of 0.5% levofloxacin ophthalmic solution for the treatment of bacterial conjunctivitis in pediatric patients. AB - PURPOSE: To compare the efficacy and safety of 0.5% levofloxacin ophthalmic solution (Quixin; Santen, Napa, CA) with 0.3% ofloxacin (Ocuflox; Allergan Inc., Irvine, CA) and placebo for the treatment of pediatric bacterial conjunctivitis. METHODS: This study was a subset analysis of 167 pediatric patients (age range, 1 to 16 years) from two randomized, double-masked, multicenter, parallel group studies. Eye drops were instilled every 2 hours on days 1 and 2 and every 4 hours on days 3 through 5. Ocular signs and symptoms were noted, and conjunctival cultures were obtained on day 1 (baseline), days 3 to 5 (interim), and days 6 to 10 (final). Endpoint was defined as the last evaluable observation. Microbial and clinical outcomes were based on culture results and cardinal signs, respectively. RESULTS: At endpoint (mean of 6.5 days for 118 evaluable patients), 0.5% levofloxacin treatment demonstrated greater microbial eradication rates (percentage of patients with absence of causative organisms cultured at baseline) compared with 0.3% ofloxacin or placebo. In children age 2 to 11 years, this finding was statistically significant in favor of 0.5% levofloxacin (87% vs 62% with 0.3% ofloxacin [P < or =.032] and 88% vs 24% with placebo [P <.001]). No significant differences between treatment groups in microbial eradication rates were noted in other age subgroups. CONCLUSIONS: After 5 days of therapy, 0.5% levofloxacin ophthalmic solution was found to be safe and effective in treating pediatric bacterial conjunctivitis. Treatment with 0.5% levofloxacin achieved microbial eradication rates in children that were statistically superior to those attained with 0.3% ofloxacin or placebo. PMID- 14566314 TI - Partial rectus muscle-augmented transpositions in abduction deficiency. AB - PURPOSE: Lateral posterior fixation sutures increase the effect of full rectus extraocular muscle transpositions. Partial rectus muscle transposition may be indicated to minimize the risk of anterior ischemia when multiple rectus muscles require surgery to achieve ocular alignment. PURPOSE: To report a modification of full vertical rectus muscle transposition with lateral posterior fixation sutures for use in patients at risk for anterior segment ischemia. METHODS: Ten cases of unilateral split rectus muscle transposition augmented with lateral posterior fixation sutures were analyzed. Five patients had Duane's syndrome with esotropia in primary position, and five patients had sixth-nerve palsy. RESULTS: Seven patients had a history of ipsilateral rectus muscle surgery, and three patients underwent simultaneous surgery on ipsilateral horizontal rectus muscles. In Duane's syndrome patients, the preoperative angle of deviation at distance was 15.8 +/- 5.8 prism diopters (PD) (range, 10 to 25) compared with 3.2 +/- 4.4 PD (range, 0 to 8) postoperatively (P =.005). In patients with sixth-nerve palsy, the preoperative angle of deviation at distance was 45.2 +/- 23.9 PD (range, 16 to 80) compared with -5 +/- 14.1 PD (range, -30 to 5) postoperatively (P =.004). Postoperative binocular single visual fields enlarged in seven of seven patients. CONCLUSION: Partial rectus muscle-augmented transposition allows surgery on multiple ipsilateral rectus muscles in (1) Duane's syndrome patients with esotropia, marked cocontraction, and/or limitation to both horizontal rotations and in (2) sixth-nerve palsy patients with ipsilateral medial rectus tightness. Augmented partial rectus muscle transpositions improve ocular alignment and may enlarge binocular single fields in patients with persistent deviations despite previous muscle surgery. PMID- 14566315 TI - Validity of the Titmus and Randot circles tasks in children with known binocular vision disorders. AB - PURPOSE: The Titmus and Randot (version 2) circles tests contain monocular form cues that may enable patients with binocular vision disorders to pass without any measurable stereopsis. The purpose of this study was to evaluate the validity of the Randot and Titmus circles tests for quantifying stereoacuity in children with known binocular vision disorders. METHODS: Participants included 170 consecutive children with diverse binocular sensory function ranging from fine to no measurable stereopsis. Stereoacuity was measured using the Randot circles, the Titmus circles, and the Randot Preschool Stereoacuity tests. RESULTS: Discrepancies between stereoacuity scores derived using either the Titmus or Randot circles tests and the Randot Preschool Stereoacuity test increased as a function of random-dot stereoacuity. Stereoacuity scores derived using the circles tests showed good agreement with random-dot stereoacuity when stereoacuity was 2.2 log seconds of arc (160 seconds of arc) or better, but they progressively overestimated stereoacuity for poorer random-dot stereoacuity scores. CONCLUSION: When measuring stereoacuity using either the Titmus or Randot circles tests in patients with known binocular vision disorders, stereoacuity scores > 2.2 log seconds of arc should be interpreted with caution because it is above this level of stereoacuity that the monocular form cues of each of the tests may invalidate the results. PMID- 14566316 TI - Congenital absence of the inferior rectus muscle--diagnosis and management. AB - BACKGROUND: Congenital absence of the inferior rectus muscle is a rare cause of apparent inferior rectus palsy especially in the absence of associated cranial facial anomalies. METHODS: We report three cases of isolated congenital absence of the inferior rectus muscle and its successful surgical management. RESULTS: Failure of the normal embryologic development of the mesodermal complex around the eye can lead to agenesis of the extraocular muscles. In apparent palsies of the inferior rectus muscle and no definite cause, a high index of suspicion and orbital imaging can confirm the diagnosis of congenitally absent inferior rectus preoperatively. Surgical correction may involve inferior transposition of the horizontal rectus muscles. CONCLUSIONS: Although rare, congenital absence of the inferior rectus muscle is a possible cause of apparent inferior rectus muscle palsy particularly in the absence of another identifiable cause. Strabismus surgery in conjunction with intramuscular botulinum toxin injection can offer significant improvement in function and cosmesis of these patients. PMID- 14566317 TI - Hydroxyapatite orbital implant exposure in children. AB - PURPOSE: Hydroxyapatite orbital implants are buried implants that may be integrated into the overlying prosthesis after enucleation. We report problems encountered using these implants during a 14-year period in a pediatric population. METHODS: Retrospective analysis of a pediatric population from 1987 through 2001. RESULTS: Indications for enucleation (N = 19) included retinoblastoma (n =17), persistent hyperplastic primary vitreous (n =1), and painful blind eye (n =1). Conjunctival erosion (36.84%) and consequent implant exposure (15.70%) were the main problems identified in this study. There were no cases of orbital infection. Management included resuturing and scleral patching. CONCLUSION: Conjunctival erosion of hydroxyapatite implants contributed to significant morbidity in 19 children who underwent enucleation and hydroxyapatite orbital implant. PMID- 14566318 TI - Progressive increase in the angle of deviation in acquired nonaccommodative esotropia of childhood. AB - PURPOSE: Although the angle of deviation in patients with congenital esotropia has been shown to increase during the early preoperative period, few data exist regarding this change in other forms of childhood esotropia. The primary objective of this study was to investigate whether the angle of deviation increases with time in patients with acquired nonaccommodative esotropia (ANAET). METHODS: The medical records of all children whose ANAET was diagnosed at East Tennessee State University College of Medicine from August 1, 1995, through October 1, 2001, were retrospectively reviewed. All patients were assessed for any change in the amount of their strabismic angle and associated risk factors. RESULTS: Eighty-two patients with ANAET were identified, and all except 3 underwent more than one examination. Of the 79 children who underwent more than one examination, 26 (32.9%) displayed an angle increase of > or =10 prism diopters (PD) during a median follow-up period of 2.9 months. The ANAET of those patients whose angle increased by > or =10 PD was diagnosed (28.4 months of age vs 45.5 months of age, P =.003), and they underwent surgery (35.9 months of age vs 53.3 months of age, P =.003) at a significantly younger median age than those whose angle increased <10 PD. CONCLUSIONS: The angle of deviation of approximately one third of patients with ANAET in this study increased at least 10 PD in the early preoperative period. This increase occurs more often in younger patients and underscores the importance of performing serial measurements of the deviating angle before surgical intervention is undertaken. PMID- 14566319 TI - Mutation of the FOXC2 gene in familial distichiasis. AB - OBJECTIVE: To examine the FOXC2 gene in a family with hereditary distichiasis. BACKGROUND: Distichiasis, ie, a second row of eyelashes arising from the meibomian glands of the eyelids, can be inherited either alone (Online Mendelian Inheritance in Man [OMIM] no. 126300) or, more commonly, as part of the lymphedema-distichiasis (LD) syndrome (OMIM no. 153400). More than 45 families with mutations in the FOXC2 gene and LD have been described. Both lymphedema and distichiasis are highly penetrant. Distichiasis without lymphedema is not commonly seen. METHODS: We examined three generations of a family (N = nine members) with hereditary distichiasis but without lymphedema or other features of LD syndrome. The FOXC2 gene was polymerase chain reaction--amplified from genomic DNA from all family members and examined for mutations. RESULTS: Clinical examination showed distichiasis of all four lids in two affected family members across two generations. There were no other consistent ophthalmologic abnormalities in the family. A cytosine-to-adenine transversion was identified in DNA from affected study participants at nucleotide position 1076, which would be predicted to cause truncation of the protein at codon 359. This change was not observed in any of the nine unaffected family members participating. CONCLUSIONS: This finding suggests that hereditary distichiasis and LD may not be separate genetic disorders but different phenotypic expressions of the same underlying disorder. Ophthalmologists should be aware that LD may present as distichiasis alone and counsel and refer their patients appropriately. PMID- 14566321 TI - Wide cavum septum pellucidum and visual impairment. PMID- 14566320 TI - Single-stage adjustable strabismus surgery for restrictive strabismus. AB - PURPOSE: To evaluate the feasibility and stability of ocular alignment of single stage adjustable strabismus surgery (SSASS) in restrictive strabismus. METHODS: This was an observational case series comprising 12 patients with restrictive strabismus (mean age, 54.8 years) who were treated with SSASS using intravenous midazolam, fentanyl, and topical anesthesia. All were studied in a retrospective institutional manner. The refractive strabismus in 7 patients was caused by dysthyroid orbitopathy. Five patients had undergone previous ocular surgery, and 4 had undergone previous strabismus surgery. SSASS typically involved the vertical rectus muscles. Horizontal rectus muscles were adjusted when necessary. Silicon-treated polyester suture material (Ti-cron; United States Surgical, Norwalk, CT no longer available), 6-0, were used for inferior rectus recessions. Ocular alignment was set at ortho at the end of surgery and evaluated at 2 days, 6 weeks, and 3 months after surgery. The typical hang-back procedure was to lock the suture at the middle and edges of the tendon or muscle at the intended disinsertion point. The tendon was then disinserted and hung back from the original insertion with adjustments until the desired position (ortho) and single vision were attained. RESULTS: All patients remained comfortable throughout surgery and had no significant postoperative discomfort. All patients except 2 (16.6%) maintained satisfactory vertical alignment (<2 prism diopters). These 2 patients with dysthyroid orbitopathy had progressive overcorrection after inferior rectus recession. CONCLUSIONS: SSASS, using intravenous midazolam, fentanyl, and topical anesthesia, is a safe and precise alternative treatment for patients with restrictive strabismus including those with dysthyroid orbitopathy. PMID- 14566322 TI - Bilateral extraocular muscle epithelial inclusion cysts as a complication of strabismus surgery. PMID- 14566323 TI - Loss of retinoblastoma but not p16 function allows bypass of replicative senescence in human fibroblasts. AB - Current models envision replicative senescence to be under dual control by the p53 and retinoblastoma (RB) tumour-suppressor pathways. The role of the p16(INK4a)-RB pathway is controversial, and the function of RB in human cells has not been tested directly. We used targeted homologous recombination to knock out one copy of RB in presenescent human fibroblasts. During entry into senescence, RB+/- cells underwent spontaneous loss of heterozygosity and the resultant RB-/- clones bypassed senescence. The extended lifespan phase was eventually terminated by a crisis-like state. The same phenotype was documented for p21(CIP1/WAF1) and p53 heterozygous cells, indicating that loss of function of all three genes results in failure to establish senescence. By contrast, the abolition of p16 function by the expression of a p16-insensitive cyclin-dependent kinase 4 protein or siRNA-mediated knockdown provided only minimal lifespan extension that was terminated by senescence. We propose that p53, p21 and RB act in a linear genetic pathway to regulate cell entry into replicative senescence. PMID- 14566324 TI - Efficient transgenesis in farm animals by lentiviral vectors. AB - Microinjection of DNA is now the most widespread method for generating transgenic animals, but transgenesis rates achieved this way in higher mammals are extremely low. To address this longstanding problem, we used lentiviral vectors carrying a ubiquitously active promoter (phosphoglycerate kinase, LV-PGK) to deliver transgenes to porcine embryos. Of the 46 piglets born, 32 (70%) carried the transgene DNA and 30 (94%) of these pigs expressed the transgene (green fluorescent protein, GFP). Direct fluorescence imaging and immunohistochemistry showed that GFP was expressed in all tissues of LV-PGK transgenic pigs, including germ cells. Importantly, the transgene was transmitted through the germ-line. Tissue-specific transgene expression was achieved by infecting porcine embryos with lentiviral vectors containing the human keratin K14 promoter (LV-K14). LV K14 transgenic animals expressed GFP specifically in basal keratinocytes of the skin. Finally, infection of bovine oocytes after and before in vitro fertilization with LV-PGK resulted in transgene expression in 45% and 92% of the infected embryos, respectively. PMID- 14566325 TI - Genome-wide distribution of DNA replication origins at A+T-rich islands in Schizosaccharomyces pombe. AB - Genome-wide analysis of replication dynamics requires the previous identification of DNA replication origins (ORIs). However, variability among the ORIs makes it difficult to predict their distribution across the genome on the basis of their sequence. We report here that ORIs in Schizosaccharomyces pombe coincide with discrete chromosomal A+T-rich islands of up to 1 kb long that are characterized by a distinctive A+T content that clearly differentiates them from the rest of the genome. Genome-wide analysis has enabled us to identify 384 of these regions, which predicts the position of most ORIs in the genome, as shown by functional replication analyses. A+T-rich islands occur at the mating locus, centromeres and subtelomeric regions at a density that is approximately fourfold higher than elsewhere in the genome, which suggests a link between the origin recognition complex (ORC) and transcriptional silencing in these regions. The absence of consensus elements in A+T-rich islands implies that different sequences can target the ORC to different ORIs. PMID- 14566326 TI - Hexameric ring structure of the full-length archaeal MCM protein complex. AB - In eukaryotes, a family of six homologous minichromosome maintenance (MCM) proteins has a key function in ensuring that DNA replication occurs only once before cell division. Whereas all eukaryotes have six paralogues, in some Archaea a single protein forms a homomeric assembly. The complex is likely to function as a helicase during DNA replication. We have used electron microscopy to obtain a three-dimensional reconstruction of the full-length MCM from Methanobacterium thermoautotrophicum. Six monomers are arranged around a sixfold axis, generating a ring-shaped molecule with a large central cavity and lateral holes. The channel running through the molecule can easily accommodate double-stranded DNA. The crystal structure of the amino-terminal fragment of MCM and a model for the AAA+ hexamer have been docked into the map, whereas additional electron density suggests that the carboxy-terminal domain is located at the interface between the two domains. The structure suggests that the MCM complex is likely to act in a different manner to traditional hexameric helicases and is likely to bear more similarity to the SV40 large T antigen or to double-stranded DNA translocases. PMID- 14566327 TI - Pre-TCR-triggered ERK signalling-dependent downregulation of E2A activity in Notch3-induced T-cell lymphoma. AB - Notch and basic helix-loop-helix E2A pathways specify cell fate and regulate neoplastic transformation in a variety of cell types. Whereas Notch enhances tumorigenesis, E2A suppresses it. However, whether and how Notch and E2A interact functionally in an integrative mechanism for regulating neoplastic transformation remains to be understood. It has been shown that Notch3-induced T-cell leukaemia is abrogated by the inactivation of pTalpha/pre-T-cell antigen receptor (pre TCR). We report here that Notch3-induced transcriptional activation of pTalpha/pre-TCR is responsible for the downregulation of E2A DNA binding and transcriptional activity. Further, the E2A messenger RNA and protein levels remain unaltered but the E2A inhibitor Id1 expression is augmented in thymocytes and T lymphoma cells derived from Notch3 transgenic mice. The increase in Id1 expression is achieved by pre-TCR-induced extracellular-signalling-regulated kinase 1/2. These observations support a model in which the upregulation of pre TCR signalling seems to be the prerequi-site for Notch3-induced inhibition of E2A, thus leading to the development of lymphoma in Notch3 transgenic mice. PMID- 14566328 TI - West Nile virus encephalitis causing fatal CNS toxicity after hematopoietic stem cell transplantation. AB - We describe here a patient who died of progressive CNS deterioration following allogeneic stem cell transplant with West Nile virus as the sole pathogen on the cerebrospinal fluid and brain tissue analysis. A 50-year-old male with Philadelphia chromosome-positive acute lymphocytic leukemia (ALL) underwent allogeneic PBSCT from his HLA identical sister. After engraftment, the patient developed fever with progressive and ultimately fatal neurological deterioration. Imaging studies of the brain including CT and MRI scans were remarkable for mild low attenuation lesions of the white matter. CSF analysis was negative for neoplastic cells, bacteria, AFB, CMV, HSV, fungal infections and leukemic relapse. However, serological analysis of both the serum and CSF was positive for West Nile virus-specific IgM antibodies. At autopsy, West Nile virus PCR and cultures were positive in the mid-brain tissue. Electron micrographs showed evidence of viral particles. Given the recent increase in the spread of West Nile virus infections and the increased susceptibility of BMT patients to infectious complications, West Nile virus encephalitis should be considered in patients undergoing transplantation. PMID- 14566329 TI - Allogeneic bone marrow transplantation vs chemotherapy for children with Philadelphia chromosome-positive acute lymphoblastic leukemia. AB - Allogeneic bone marrow transplant (BMT) with an MRD in complete remission (CR)1 is the preferred treatment for children with Philadelphia-positive (Ph(+)) ALL. The role of MUD BMT in CR1 is still controversial. We compared the outcomes of two treatment strategies: BMT using an MRD or MUD vs chemotherapy in children with Ph(+) ALL in CR1. In total, 21 children were treated from 1985 to 2001. In all, 10 received chemotherapy and 11 received allogeneic BMT: four MRD, seven MUD. In the MRD group, one relapsed 12 months after BMT and died; the remaining three are long-term event-free survivors (median follow-up, 6.1 years). In the MUD group four died; the remaining three are long-term event-free survivors (median follow-up, 7.2 years). The 4-year event-free survival (EFS) for the BMT group was 53+/-15%. In the chemotherapy group, seven relapsed after a median period of 12.5 months and three remain in continuous CR (median follow-up, 2.4 years). Four chemotherapy patients received CR2 transplants; all died. The 4-year EFS for the chemotherapy and MUD groups was 33+/-17 and 35.7+/-20%, respectively. This difference was not statistically significant. We continue to support treating children with Ph(+) ALL with MRD BMT in CR1. The effectiveness of MUD BMT vs chemotherapy merits further study. PMID- 14566330 TI - Alemtuzumab can successfully treat steroid-refractory acute graft-versus-host disease (aGVHD). PMID- 14566331 TI - Incorporation of aminoacyl-tRNA into the ribosome as seen by cryo-electron microscopy. AB - Aminoacyl-tRNAs (aa-tRNAs) are delivered to the ribosome as part of the ternary complex of aa-tRNA, elongation factor Tu (EF-Tu) and GTP. Here, we present a cryo electron microscopy (cryo-EM) study, at a resolution of approximately 9 A, showing that during the incorporation of the aa-tRNA into the 70S ribosome of Escherichia coli, the flexibility of aa-tRNA allows the initial codon recognition and its accommodation into the ribosomal A site. In addition, a conformational change observed in the GTPase-associated center (GAC) of the ribosomal 50S subunit may provide the mechanism by which the ribosome promotes a relative movement of the aa-tRNA with respect to EF-Tu. This relative rearrangement seems to facilitate codon recognition by the incoming aa-tRNA, and to provide the codon anticodon recognition-dependent signal for the GTPase activity of EF-Tu. From these new findings we propose a mechanism that can explain the sequence of events during the decoding of mRNA on the ribosome. PMID- 14566332 TI - Nanoscale effects leading to non-Einstein-like decrease in viscosity. AB - Nanoparticles have been shown to influence mechanical properties; however, transport properties such as viscosity have not been adequately studied. This might be due to the common observation that particle addition to liquids produces an increase in viscosity, even in polymeric liquids, as predicted by Einstein nearly a century ago. But confinement and surface effects provided by nanoparticles have been shown to produce conformational changes to polymer molecules, so it is expected that nanoparticles will affect the macroscopic viscosity. To minimize extraneous enthalpic or other effects, we blended organic nanoparticles, synthesized by intramolecular crosslinking of single polystyrene chains, with linear polystyrene macromolecules. Remarkably, the blend viscosity was found to decrease and scale with the change in free volume introduced by the nanoparticles and not with the decrease in entanglement. Indeed, the entanglements did not seem to be affected at all, suggesting unusual polymer dynamics. PMID- 14566333 TI - PML-RARA-targeted DNA vaccine induces protective immunity in a mouse model of leukemia. AB - Despite improved molecular characterization of malignancies and development of targeted therapies, acute leukemia is not curable and few patients survive more than 10 years after diagnosis. Recently, combinations of different therapeutic strategies (based on mechanisms of apoptosis, differentiation and cytotoxicity) have significantly increased survival. To further improve outcome, we studied the potential efficacy of boosting the patient's immune response using specific immunotherapy. In an animal model of acute promyelocytic leukemia, we developed a DNA-based vaccine by fusing the human promyelocytic leukemia-retinoic acid receptor-alpha (PML-RARA) oncogene to tetanus fragment C (FrC) sequences. We show for the first time that a DNA vaccine specifically targeted to an oncoprotein can have a pronounced effect on survival, both alone and when combined with all-trans retinoic acid (ATRA). The survival advantage is concomitant with time-dependent antibody production and an increase in interferon-gamma (IFN-gamma). We also show that ATRA therapy on its own triggers an immune response in this model. When DNA vaccination and conventional ATRA therapy are combined, they induce protective immune responses against leukemia progression in mice and may provide a new approach to improve clinical outcome in human leukemia. PMID- 14566334 TI - An animal model of age-related macular degeneration in senescent Ccl-2- or Ccr-2 deficient mice. AB - The study and treatment of age-related macular degeneration (AMD), a leading cause of blindness, has been hampered by a lack of animal models. Here we report that mice deficient either in monocyte chemoattractant protein-1 (Ccl-2; also known as MCP-1) or its cognate C-C chemokine receptor-2 (Ccr-2) develop cardinal features of AMD, including accumulation of lipofuscin in and drusen beneath the retinal pigmented epithelium (RPE), photoreceptor atrophy and choroidal neovascularization (CNV). Complement and IgG deposition in RPE and choroid accompanies senescence in this model, as in human AMD. RPE or choroidal endothelial production of Ccl-2 induced by complement C5a and IgG may mediate choroidal macrophage infiltration into aged wild-type choroids. Wild-type choroidal macrophages degrade C5 and IgG in eye sections of Ccl2(-/-) or Ccr2(-/ ) mice. Impaired macrophage recruitment may allow accumulation of C5a and IgG, which induces vascular endothelial growth factor (VEGF) production by RPE, possibly mediating development of CNV. These models implicate macrophage dysfunction in AMD pathogenesis and may be useful as a platform for validating therapies. PMID- 14566335 TI - CD46 is a cellular receptor for group B adenoviruses. AB - Group B adenoviruses, a subgenus of human Adenoviridae, are associated with a variety of often-fatal illnesses in immunocompromised individuals, including bone marrow transplant recipients and cancer and AIDS patients. Recently, group B adenovirus derivatives have gained interest as attractive gene therapy vectors because they can transduce target tissues, such as hematopoietic stem cells, dendritic cells and malignant tumor cells, that are refractory to infection by commonly used adenoviral vectors. Whereas many adenoviruses infect cells through the coxsackievirus and adenovirus receptor (CAR), group B adenoviruses use an alternate, as-yet-unidentified cellular attachment receptor. Using mass spectrometric analysis of proteins interacting with a group B fiber, we identified human CD46 as a cellular attachment receptor for most group B adenoviruses. We show that ectopic expression of human CD46 rendered nonhuman cells susceptible to infection with group B viruses in vitro and in vivo. In addition, both siRNA-mediated knockdown of CD46 and a soluble form of CD46 blocked infection of human cell lines and primary human cells. The discovery that group B adenoviruses use CD46, a ubiquitously expressed complement regulatory protein, as a cellular attachment receptor elucidates the diverse clinical manifestation of group B virus infections, and bears directly on the application of these vectors for gene therapy. PMID- 14566336 TI - Adaptor protein 3-dependent microtubule-mediated movement of lytic granules to the immunological synapse. AB - Hermansky-Pudlak syndrome (HPS) is a rare autosomal recessive disease characterized by platelet defects and oculocutaneous albinism. Individuals with HPS type 2 (HPS2) lack the cytosolic adaptor protein 3 (AP-3) involved in lysosomal sorting, and are also immunodeficient. Here we characterize an HPS2 mutation and demonstrate that AP-3 deficiency leads to a loss of cytotoxic T lymphocyte (CTL)-mediated cytotoxicity. Although the lysosomal protein CD63 was mislocalized to the plasma membrane, perforin and granzymes were correctly localized to the lytic granules in AP-3-deficient CTLs. However, the lytic granules of AP-3-deficient CTLs were enlarged and were unable to move along microtubules and dock within the secretory domain of the immunological synapse. These data show that AP-3 is essential for polarized secretion from CTLs. PMID- 14566337 TI - Control of dendritic cell cross-presentation by the major histocompatibility complex class I cytoplasmic domain. AB - Dendritic cells (DCs) can present extracellularly derived antigens in the context of major histocompatibility complex (MHC) class I molecules, a process called cross-presentation. Although recognized to be important for priming of T cell responses to many viral, bacterial and tumor antigens, the mechanistic details of this alternative antigen-presentation pathway are poorly understood. We demonstrate here the existence of an endolysosomal compartment in DCs where exogenously derived peptides can be acquired for presentation to T cells, and show that the MHC class I cytoplasmic domain contains a tyrosine-based targeting signal required for routing MHC class I molecules through these compartments. We also report that transgenic mice expressing H-2K(b) with a tyrosine mutation mount inferior H-2K(b)-restricted cytotoxic T lymphocyte responses against two immunodominant viral epitopes, providing evidence of a crucial function for cross priming in antiviral immunity. PMID- 14566339 TI - DNA helicase gene interaction network defined using synthetic lethality analyzed by microarray. AB - We describe a new synthetic lethality analysis by microarray (SLAM) technique that uses approximately 4,600 Saccharomyces cerevisiae haploid deletion mutants with molecular 'bar codes' (TAGs). We used SGS1 and SRS2, two 3'-->5' DNA helicase genes, as 'queries' to identify their redundant and unique biological functions. We introduced these 'query mutations' into a haploid deletion pool by integrative transformation to disrupt the query gene in every cell, generating a double mutant pool. Optimization of integrative transformation efficiency was essential to the success of SLAM. Synthetic interactions defined a DNA helicase genetic network and predicted a role for SRS2 in processing damaged replication forks but, unlike SGS1, not in rDNA replication, DNA topology or lagging strand synthesis. SGS1 and SRS2 have synthetic defects with MRC1 but not RAD9, suggesting that SGS1 and SRS2 function in a parallel pathway with MRC1 to transduce the DNA replication stress signal to the general DNA damage checkpoint pathway. Both helicase genes have rad51-reversible synthetic defects with 5'-->3' DNA helicase RRM3, suggesting that RRM3 helps prevent formation of toxic recombination intermediates. SLAM detects synthetic lethality efficiently and ranks candidate genetic interactions, making it an especially useful method. PMID- 14566338 TI - Positional cloning of a novel gene influencing asthma from chromosome 2q14. AB - Asthma is a common disease in children and young adults. Four separate reports have linked asthma and related phenotypes to an ill-defined interval between 2q14 and 2q32 (refs. 1-4), and two mouse genome screens have linked bronchial hyper responsiveness to the region homologous to 2q14 (refs. 5,6). We found and replicated association between asthma and the D2S308 microsatellite, 800 kb distal to the IL1 cluster on 2q14. We sequenced the surrounding region and constructed a comprehensive, high-density, single-nucleotide polymorphism (SNP) linkage disequilibrium (LD) map. SNP association was limited to the initial exons of a solitary gene of 3.6 kb (DPP10), which extends over 1 Mb of genomic DNA. DPP10 encodes a homolog of dipeptidyl peptidases (DPPs) that cleave terminal dipeptides from cytokines and chemokines, and it presents a potential new target for asthma therapy. PMID- 14566340 TI - Microsomal prostaglandin E synthase-1 is the central switch during immune-induced pyresis. AB - We studied the febrile response in mice deficient in microsomal prostaglandin E synthase-1 (mPGES-1), an inducible terminal isomerase expressed in cytokine sensitive brain endothelial cells. These animals showed no fever and no central prostaglandin (PG) E2 synthesis after peripheral injection of bacterial-wall lipopolysaccharide, but their pyretic capacity in response to centrally administered PGE2 was intact. Our findings identify mPGES-1 as the central switch during immune-induced pyresis and as a target for the treatment of fever and other PGE2-dependent acute phase reactions elicited by the brain. PMID- 14566341 TI - Speed and accuracy of olfactory discrimination in the rat. AB - The sense of smell is typically thought of as a 'slow' sense, but the true temporal constraints on the accuracy of olfactory perception are not known. It has been proposed that animals make finer odor discriminations at the expense of additional processing time. To test this idea, we measured the relationship between the speed and accuracy of olfactory discrimination in rats. We found that speed of discrimination was independent of odor similarity, as measured by overlap of glomerular activity patterns. Even when pushed to psychophysical limits using mixtures of two odors, rats needed to take only one sniff (<200 ms at theta frequency) to make a decision of maximum accuracy. These results show that, for the purpose of odor quality discrimination, a fully refined olfactory sensory representation can emerge within a single sensorimotor or theta cycle, suggesting that each sniff can be considered a snapshot of the olfactory world. PMID- 14566342 TI - Regulation of gene expression and cocaine reward by CREB and DeltaFosB. AB - DeltaFosB (a truncated form of FosB) and CREB (cAMP response element binding protein) are transcription factors induced in the brain's reward pathways after chronic exposure to drugs of abuse. However, their mechanisms of action and the genes they regulate remain unclear. Using microarray analysis in the nucleus accumbens of inducible transgenic mice, we found that CREB and a dominant negative CREB have opposite effects on gene expression, as do prolonged expression of DeltaFosB and the activator protein-1 (AP-1) antagonist DeltacJun. However, unlike CREB, short-term and prolonged DeltaFosB induction had opposing effects on gene expression. Gene expression induced by short-term DeltaFosB and by CREB was strikingly similar, and both reduced the rewarding effects of cocaine, whereas prolonged DeltaFosB expression increased drug reward. Gene expression after a short cocaine treatment was more dependent on CREB, whereas gene expression after a longer cocaine treatment became increasingly DeltaFosB dependent. These findings help define the molecular functions of CREB and DeltaFosB and identify clusters of genes that contribute to cocaine addiction. PMID- 14566343 TI - Olfactomotor activity during imagery mimics that during perception. AB - Neural representations created in the absence of external sensory stimuli are referred to as imagery, and such representations may be augmented by reenactment of sensorimotor processes. We measured nasal airflow in human subjects while they imagined sights, sounds and smells, and only during olfactory imagery did subjects spontaneously enact the motor component of olfaction--that is, they sniffed. Moreover, as in perception, imagery of pleasant odors involved larger sniffs than imagery of unpleasant odors, suggesting that the act of sniffing has a functional role in creating of olfactory percepts. PMID- 14566344 TI - A functional genetic variation of the 5-HT2a receptor affects human memory. AB - Human memory capacity is highly variable across individuals and is influenced by both genetic and environmental factors. A roughly 50% heritability estimate indicates that naturally occurring genetic variations have an important impact on this cognitive ability. Therefore, we investigated a functional variation of a memory-related serotonin receptor in 349 healthy young volunteers, and found 21% poorer memory performance in subjects with the rare variant. PMID- 14566345 TI - Immunofluorescence stimulated emission depletion microscopy. AB - We report immunofluorescence imaging with a spatial resolution well beyond the diffraction limit. An axial resolution of approximately 50 nm, corresponding to 1/16 of the irradiation wavelength of 793 nm, is achieved by stimulated emission depletion through opposing lenses. We have demonstrated not only that an antibody tagged label is stable enough to be recorded in this microscopy mode, but also that subdiffraction resolution can be obtained using a standard immunofluorescence preparation. PMID- 14566346 TI - Hematologic abnormalities in severe neonatal necrotizing enterocolitis: 25 years later. AB - Necrotizing enterocolitis (NEC), the most common surgical emergency in newborns, remains a therapeutic challenge for clinicians. The hematological manifestations associated with NEC were first described 25 years ago. This review discusses current knowledge of the pathophysiology involved in disturbances in megakaryocytopoiesis, coagulation, leukopoiesis, and erythropoiesis that accompany the clinical entity NEC. The discussion includes current understanding of and potential strategies for treating the hematopoietic disturbances that occur secondary to NEC. PMID- 14566347 TI - Comparison of three outpatient regimens in the management of nausea and vomiting in pregnancy. AB - OBJECTIVE: This study compares pyridoxine-metoclopramide combination therapy to prochlorperazine and promethazine monotherapies in the outpatient treatment of nausea and vomiting in pregnancy. STUDY DESIGN: In total, 174 first trimester, singleton pregnancies were evaluated for nausea and vomiting. Patients were prospectively randomized into three treatment groups: pyridoxine-metoclopramide, prochlorperazine, or promethazine. Prior to, and on the third day, patients recorded their subjective responses to the given treatment and their number of emesis episodes. The three treatment groups were compared for therapy response. RESULTS: There were no differences in the number of emesis episodes prior to treatment. Both subjective and objective responses to treatment differed among the three groups when comparing the combination therapy to the monotherapies (p<0.05). CONCLUSION: Combination therapy with pyridoxine and metoclopramide appears to be superior to either monotherapy in the treatment of nausea and vomiting in pregnancy. PMID- 14566348 TI - Prediction of fetal anemia by middle cerebral artery peak systolic velocity in pregnancies complicated by rhesus isoimmunization. AB - OBJECTIVES: To study the correlation of peak systolic velocity in the middle cerebral artery with hemoglobin concentration in fetuses at risk of anemia due to Rhesus isoimmunization. DESIGN: Peak systolic velocity of middle cerebral artery (MCA-PSV) was measured before 66 cordocentesis procedures in 20 isoimmunized fetuses. Reference values were derived from a study of 300 control fetuses. MCA PSV values and hemoglobin concentrations were expressed as multiples of the median (MoM) for gestational age. The following hemoglobin concentration MoM thresholds defined degrees of anemia: mild, between 0.83 and 0.65; moderate, between 0.64 and 0.55; and severe, less than 0.55. Regression analysis was performed and receiver-operator-characteristic curves were constructed to determine the diagnostic accuracy of different thresholds of MCA-PSV for the prediction of moderate to severe anemia, either at the initial or repeat cordocentesis procedures. RESULTS: The mean (+/-SD) gestational age at cordocentesis was 28.5+/-4.6 weeks. Moderate to severe anemia was observed on 29 (44%) and hydrops on 27 (41%) occasions. MCA-PSV correlated weakly with hemoglobin concentrations. At threshold values 1.50 MoM, the sensitivity, specificity, and negative predictive value for moderate to severe anemia were 9.0, 100, and 48.0% at the initial cordocentesis procedures, and 44.0, 96.0, and 73.0% at repeat cordocentesis procedures, respectively. CONCLUSIONS: Although MCA PSV is highly specific, negative values do not rule out fetal anemia. Further research is required before it can be recommended in clinical practice. PMID- 14566349 TI - The value of Ureaplasma urealyticum tracheal culture and treatment in premature infants following an acute respiratory deterioration. AB - OBJECTIVE: To determine if treatment of Ureaplasma urealyticum (Uu), found at the time of an acute respiratory deterioration, decreases the incidence of chronic lung disease (CLD) in very low birth weight infants (VLBW). STUDY DESIGN: Between 1996 and 1999, medical records of all mechanically ventilated VLBW infants, who had an acute respiratory deterioration, were reviewed for gestational age (GA), birth weight (BW), gender, presence of CLD, Uu tracheal cultures, and erythromycin treatment. RESULTS: A total of 100 patients met our inclusion criteria (GA: 26.2+/-1.7 weeks, BW: 737+/-167.1 g (mean+/-SD)). Uu was present in 46.3% (38/82) of patients with CLD versus 50% (9/18) of patients without CLD (odds ratio 0.86 (CI: 0.31 to 2.39); p=0.77). Erythromycin treatment was not found to be protective against the development of CLD (odds ratio: 1.46 (CI: 0.25 to 8.31); p=0.66). CONCLUSION: Following an acute respiratory deterioration, tracheal isolation, and treatment of Uu may not decrease the incidence of CLD in VLBW infants. PMID- 14566350 TI - Mean arterial pressure in very low birth weight (801 to 1500 g) concordant and discordant twins during the first day of life. AB - OBJECTIVE: To determine retrospectively mean arterial pressure (MAP) for stable and unstable concordant and discordant very low birth weight (VLBW: 801 to 1500 g) twins during the first 24 hours of life. BACKGROUND: Morbidity and mortality are much higher for extremely low birth weight (ELBW < or =800 g) than for VLBW twins. Recently, we reported MAP trends and reference values in concordant and discordant ELBW twins. No comparable information is available for VLBW infants. DESIGN: Retrospective cohort study. METHODS: We studied 48 sets of concordant and 40 sets of discordant (birth weight difference > or =20%) consecutively born VLBW twins. Stable patients were defined as having umbilical cord hemoglobin > or =14 g/dl, nonacidotic blood gases, never treated for hypotension and survived at least 7 days. MAPs (Torr) were measured by oscillometry in 3163 and by transducer via umbilical artery in 2028 instances. RESULTS: Concordant and discordant twins were similar in demographics, history of twin-twin transfusion (TTTX), antenatal steroids, chorioamnionitis, pre-eclampsia, cesarean delivery, cord hemoglobin, normal head ultrasounds or I to II intracranial hemorrhage (97 and 99%) and neonatal mortality (4 and 5%), but were different in incidence of preterm labor (83 and 58%), birth weight (1227 and 1509 g) and gestational age (GA) (30 and 32 weeks). In all, 66 (69%) concordant twins and 61 (76%) discordant twins were stable. Stable concordant twins, whether small or large, had comparable MAP on admission that increased to 24 hours. Twins of < or =32 weeks GA had lower MAP throughout than those of > or =33 weeks GA. Although their mean birth weights were similar (1262 and 1274 g), 23 stable concordant males had significantly higher MAP than 43 concordant females. Stable discordant twins were divided into 31 small (1241 g) and 30 large (1845 g); their MAPs were different (p<0.05): 35 and 39 (admission), 35 and 39 (1 hour), 36 and 46 (6 hours), 38 and 41 (12 hours), 40 and 41 (18 hours) and 42 and 42 (24 hours) Torr. In all, 88% of small discordant twins were IUGR and 91% of large discordant twins had normal growth. TTTX syndrome occurred in 12 monochorionic sets. Nine of 12 donors were IUGR while 10 of 12 recipients had normal growth. Four of 12 donors had grades III to IV intracranial hemorrhage, eight donors and all 12 recipients had normal ultrasounds. Although their cord hemoglobin levels were similar, donor and recipient MAPs were higher than in any other group and, opposite to concordant and discordant twins, their values decreased from birth to 24 hours. CONCLUSION: In stable concordant, stable discordant, and small and large discordant twins, MAP correlates with birth weight, GA and postnatal age, and increases during the first 24 hours. In recipient and donor twin-twin transfusion infants, MAP is higher throughout and declines over time. PMID- 14566351 TI - Effect of dexamethasone therapy on serum vitamin E concentrations in premature infants with bronchopulmonary dysplasia. AB - OBJECTIVE: To investigate the effect of dexamethasone therapy on serum vitamin E concentrations in premature infants with bronchopulmonary dysplasia. STUDY DESIGN: A total of 10, 24 to 29 weeks' gestational age, infants enrolled in a prospective study designed to evaluate the effect of dexamethasone on lipid intolerance were eligible for the study. Eight of these 10 infants had serum vitamin E concentrations measured simultaneously with serum triglyceride concentrations before the start of dexamethasone therapy (baseline) and within 5 days of the initiation of dexamethasone therapy. Charts were reviewed for vitamin E intake at baseline and on dexamethasone therapy for each of these eight infants. RESULTS: All eight infants had physiological serum vitamin E concentrations (1 to 3 mg/dl) at baseline, while six of eight infants had pharmacological serum vitamin E concentrations (> or =3 mg/dl) on dexamethasone therapy. All infants with an increase in serum vitamin E concentration also had a simultaneous increase in serum triglyceride concentrations with a significant correlation between vitamin E and triglyceride concentrations (Spearman's rho=0.92). There was a significant difference in mean serum vitamin E concentration between baseline and post-dexamethasone therapy (P=0.01, Wilcoxon's signed-rank test). There was no significant difference in vitamin E intake between baseline and post-dexamethasone therapy. CONCLUSION: Dexamethasone therapy in premature infants induces significant increase in serum vitamin E concentrations to pharmacological levels independent of vitamin E intake. PMID- 14566352 TI - Withdrawal of inhaled nitric oxide from nonresponders after short exposure. AB - INTRODUCTION: Inhaled nitric oxide has been approved by FDA for treatment of hypoxic respiratory failure in term and near-term neonates. The safety of withdrawing NO in patients who fail to respond is a major concern. OBJECTIVE: To obtain further insight into the safety of withdrawing NO in patients who do not initially respond to this therapy, the charts of patients enrolled at our center in the Neonatal Nitric Oxide Study (NINOS) were reviewed. Oxygenation indices (OI) before, during and after exposure to NO or placebo were compared. METHODS: The charts of 110 neonates who received NO or placebo within the NINOS trial guidelines from 1995 to 1999 were reviewed. Arterial blood gases of nonresponders before, during and after a 30-minute exposure to different doses of NO or placebo were analyzed. RESULTS: For both high-dose and 20 ppm nonresponders who received NO, there was no significant change in OI from baseline after withdrawing NO. For patients receiving placebo, there was a significant increase in OI following a 30 minute exposure to placebo. CONCLUSIONS: NO has been shown to be effective in the treatment of neonatal respiratory failure; however, not all recipients demonstrate clinical improvement. This study showed that withdrawing NO in nonresponders did not cause increased respiratory morbidity when NO exposure was limited to 30 min. Differences between our results and others showing refractory hypoxia after withdrawing NO may be due to different durations of NO exposure and, possibly, its effect on endogenous NO metabolism. PMID- 14566353 TI - Gentamicin pharmacokinetics in term newborn infants receiving high-frequency oscillatory ventilation or conventional mechanical ventilation: a case-controlled study. AB - OBJECTIVE: To compare the pharmacokinetics of gentamicin in infants receiving high-frequency oscillatory ventilation (HFOV) with infants receiving conventional mechanical ventilation. DESIGN: A case-controlled study design was used to compare the pharmacokinetics of gentamicin in critically ill infants receiving HFOV and conventional mechanical ventilation. Medical records of all full-term newborn infants (> or =37 weeks gestational age) who received either high frequency mechanical ventilation or conventional mechanical ventilation between 1991 and 2001 were reviewed and relevant patient demographics, renal function tests and gentamicin administration and plasma concentration data collected. Elimination rate constant, half-life, volume of distribution and clearance for both groups were calculated using standard kinetics equations. SETTING: A tertiary care children's hospital. PATIENTS: Newborn infants, > or =37 weeks gestational age, receiving gentamicin and high-frequency mechanical ventilation or conventional mechanical ventilation. MEASUREMENTS AND MAIN RESULTS: In total, 18 patients were included in the conventional mechanical ventilation group and 15 in the HFOV group. The mean gentamicin dose for conventional mechanical ventilation and HFOV groups infants were 2.52+/-0.07 and 2.5+/-0.07 mg/kg/dose, respectively. Initial dosing interval was 12 hours in all of the conventional mechanical ventilation infants and 13 of the 15 HFOV infants. The dosing interval for the remaining two HFOV infants was 18 hours. No patient in either group demonstrated oliguria. Statistical analysis using the Student t-test for unequal variances yielded significant differences between the two groups with regard to elimination rate constant, half-life, volume of distribution and clearance, with a p value of <0.05 for all the observations. The mean of the highest P(aw) received by each patient in the HFOV group (19.2+/-4.05) was considerably higher than in the conventional mechanical ventilation group (13.4+2.23) (p>0.05). CONCLUSION: Infants receiving HFOV had reduced gentamicin clearance. Full-term infants receiving HFOV should be initiated at gentamicin dosing intervals of 18 hours rather than the traditional 12 hours recommended for this age group. PMID- 14566354 TI - Treatment of chylothorax in a premature infant using somatostatin. AB - We describe a case of spontaneous chylothorax in a premature infant successfully treated by octreotide after other forms of conservative therapy were not effective. PMID- 14566355 TI - Bilateral cataracts, retinal detachment and vitreous hemorrhage in a newborn with congenital diaphragmatic hernia. AB - Congenital diaphragmatic hernia (CDH) is associated with a wide range of other malformations. We describe a patient with CDH who also had significant eye findings noted at birth. PMID- 14566358 TI - Gene therapy progress and prospects: gene therapy for severe combined immunodeficiency. AB - Severe combined immunodeficiencies have long been targeted as a group of disorders amenable to gene therapy because of their defined molecular biology and pathophysiology, and the prediction that corrected cells would have profound growth and survival advantage. Recently, several clinical studies have shown that conventional gene transfer technology can produce major beneficial therapeutic effects in these patients, but, as for all cellular and pharmacological treatment approaches, with a finite potential for toxicity. PMID- 14566356 TI - Laryngeal lymphatic malformation in a newborn. AB - Stridor in a neonate evokes a series of prompt evaluations to determine the etiology. Common etiologies include inherent structural defects (such as laryngomalacia), vocal fold paralysis, acquired infectious etiologies, or extrinsic compressions. We report a neonate with a lymphatic malformation (lymphangioma) initially confined to the larynx that presented with stridor and progressive respiratory failure. Only seven other cases have been reported to present with lymphatic malformation confined solely to the larynx. There are several different modalities available to treat this condition, but recurrences are common. The presentation and management are discussed below. Physicians should be aware that stridor in a neonate can result from this unusual intrinsic obstruction of the larynx. PMID- 14566359 TI - Enhanced intracellular availability and survival of hammerhead ribozymes increases target ablation in a cellular model of osteogenesis imperfecta. AB - Antisense hammerhead ribozymes have the capability to cleave complementary RNA in a sequence-dependent manner. In osteogenesis imperfecta, a genetic disorder of connective tissue, mutant collagen type I has been shown to participate in but not sustain formation of the triple helix. Selective ablation of mutant collagen gene transcript could potentially remove the mutant gene product and reverse the dominant-negative effect exerted by the abnormal protein. In earlier studies we showed that the hammerhead ribozyme Col1A1Rz547 selectively cleaved a mutant Col1A1 gene transcript in a murine calvarial osteoblast cell line. In order to test the possible therapeutic efficacy of this approach, a dramatic downregulation of the mutant transcript must be achieved, a function directly related to high steady-state level of intracellular ribozyme. We report significantly enhanced expression of Col1A1Rz547 by vaccinia T7 polymerase following infection with an attenuated T7-pol vaccinia virus as shown both by the intracellular level of the ribozyme and the cleavage of the mutant Col1A1 gene transcript. We also describe the engineering of a multimeric ribozyme construct comprising eight subunits, which can self-cleave to monomers. These studies suggest the potential use of multimeric ribozymes expressed by a vaccinia-based system in the therapy of a variety of disorders. PMID- 14566360 TI - Ex vivo gene therapy in autologous bone marrow stromal stem cells for tissue engineered maxillofacial bone regeneration. AB - This study examines the clinical relevance of tissue engineering integrating gene therapy and polymer science to bone regeneration. Bilateral maxillary defects (3 x 1.2 cm(2)) in 20 miniature swine were bridged with a bioresorbable internal splint. Constructs were created using ex vivo adenovirus bone morphogenetic protein (BMP)-2-mediated gene transfer to the expanded bone marrow mesenchymal stem cells (MSCs) 7 days before implantation. Controls were performed using adenovirus beta-galactosidase. The BMP-2 cell/construct displayed white solid bone formation after 3 months. Meanwhile, the hematoxylin and eosin and Von Kossa stains demonstrated exhibited mature woven bone with good mineralization. Additionally, three-dimensional computer tomography imaging revealed a nearly complete infraorbital rim repair. Quantitative analysis demonstrated a significant difference (P<0.001) in bone formation. Finally, biomechanical testing revealed no statistically significant difference in the maximal compressive strength of new bone formed by BMP-2 cell constructs and the normal maxilla. The data evidenced de novo bone formation capable of sustaining axial compressive loads. The measurement results showed that ex vivo replication defective adenovirus-mediated human BMP-2 gene transfer to MSCs enhances autologous bone formation in the repair of maxillary defects. PMID- 14566361 TI - Creation of immune 'stealth' genes for gene therapy through fusion with the Gly Ala repeat of EBNA-1. AB - A major obstacle in gene-therapy protocols is T-cell-mediated destruction of transgene-expressing cells. Therefore new approaches are needed to prevent rapid clearance of transduced cells. We exploited the Gly-Ala repeat (GAr) domain of the Epstein-Barr virus nuclear antigen-1, since the GAr prevents cytotoxic T lymphocyte-epitope generation. Here we show that three different enzymes (viz. the E. coli LacZ gene encoded beta-galactosidase, firefly luciferase, and HSV1 thymidine kinase) fused with the GAr retained their function. Moreover, linking GAr with beta-galactosidase successfully prevented recognition of GAr-LacZ expressing cells by beta-galactosidase-specific CTL. Nonetheless, vaccination with a GAr-LacZ adenovirus or with an allogeneic cell line expressing GAr-LacZ resulted in the induction of beta-gal-specific CTL. This demonstrates that the GAr domain does not inhibit cross presentation of antigens, but only affects breakdown of endogenously synthesized proteins. These data demonstrate how the GAr domain can be exploited to create immuno'stealth' genes by hiding transgene products from CTL-mediated immune attack. PMID- 14566362 TI - The contralateral effect conferred by intra-articular adenovirus-mediated gene transfer of viral IL-10 is specific to the immunizing antigen. AB - We have demonstrated previously that local, adenoviral-mediated gene transfer of vIL-10 to a single joint of rabbits and mice with experimental arthritis can suppress disease in both the treated and untreated contralateral joints. These therapeutic effects observed in distant untreated joints following local intra articular gene delivery have been termed the 'contralateral effect'. To begin to understand the underlying immunologic mechanism that confers this effect, a dual antigen model of antigen-induced arthritis (AIA) in rabbit knee joints was utilized. Rabbits were immunized against two antigens, ovalbumin and keyhole limpet hemocyanin, and AIA generated by intra-articular injection of each antigen into contralateral knees. Intra-articular adenovirus-mediated gene transfer of vIL-10 significantly reduced intra-articular leukocytosis and cartilage matrix degradation, while preserving near normal levels of cartilage matrix synthesis within treated joints. However, no antiarthritic effect was conferred in the contralateral control joints that received only a marker gene, in contrast to the results seen in a single-antigen AIA model. These results suggest that the distant antiarthritic effects associated with local gene delivery to joints are antigen-specific, and not due to vIL-10-induced generalized immunosuppression of the animal. PMID- 14566363 TI - Bactofection of mammalian cells by Listeria monocytogenes: improvement and mechanism of DNA delivery. AB - Bacteria-mediated transfer of plasmid DNA into mammalian cells (bactofection) is a potent approach to express plasmid-encoded heterologous proteins (protein antigens, toxins or enzymes) in a large set of different cell types including phagocytic and nonphagocytic mammalian cells. Previously, we have described a Listeria monocytogenes-mediated DNA delivery system, which releases plasmid DNA directly into the cytosol of mammalian cells by partial self-destruction of the carrier bacteria. Here we report on a second generation of this phage lysin supported bactofection system, which is greatly improved with respect to plasmid stability, transfer efficacy and biosafety. In this case, DNA release is initiated by spontaneous bacterial lysis in the infected cells cytosol which is subsequently enhanced by the simultaneously released phage lysin produced by the intracellular carrier bacteria. Bacteria that are capable of cell-to-cell spread are found to be much more efficient in bactofection than their non spreading counterparts. PMID- 14566364 TI - Specific HIV-1 env gene silencing by small interfering RNAs in human peripheral blood mononuclear cells. AB - RNA interference (RNAi) is triggered by the presence of a double-stranded RNA (dsRNA) in the cell, and results in the silencing of homologous gene expression by the specific degradation of an mRNA containing the same sequence. dsRNA mediated RNAi can be used in a wide variety of eucaryotes to induce the sequence specific inhibition of gene expression. Synthetic 21-23 nucleotide (nt) small interfering RNAs (siRNAs) with 2-nt 3' overhangs were recently found to mediate efficient sequence-specific mRNA degradation in mammalian cells. Here, we show that synthetic siRNAs targeted against the viral structural Env proteins encoded by HIV-1 can specifically suppress the expression of HIV-1 genes. The siRNA mediated RNAi also had advantages over antisense RNA-mediated inhibition, in terms of both the ease of designing effective antiviral agents and their potency. Especially, our best env-specific siRNAs, E7145 targeted to the central region of the V3 loop and E7490 targeted to the CD4 binding site of conserved regions on gp120, significantly inhibited the HIV-1 gene expression. Furthermore, E7145 and E7490 were effective against HIV-1(NL4-3) replication in PBMCs for a relatively long time (14 days). Therefore, the use of synthetic siRNAs provides a simple, rapid, and cost-effective tool for new anti-HIV-1 gene therapeutics. PMID- 14566365 TI - The JMLA readership survey. PMID- 14566366 TI - Professional boundaries and medical records management. PMID- 14566367 TI - Formation of Canadian health sciences library associations. PMID- 14566368 TI - Business as usual? PMID- 14566369 TI - Delivering Internet health resources to an underserved health care profession: school nurses. AB - PURPOSE: This paper reports on a course developed for school nurses. The course focused on locating reliable and high-quality medical information resources on the Internet. SETTING/PARTICIPANTS/RESOURCES: The Health Sciences Library System (HSLS) of the University of Pittsburgh formed a partnership with the Pennsylvania Association of School Nurses and Practitioners (PASNAP). Through this partnership, a hands-on course was offered at the PASNAP annual conference. BRIEF DESCRIPTION: As one component of the Health Information for the Public Project, a subcontract of the National Library of Medicine, HSLS collaborated with PASNAP. This collaboration resulted in HSLS librarians' offering a course titled "Access to Electronic Health Information for School Nurses" at PASNAP's annual conference. This paper describes the school nurse population, their professional information needs, and the development of the course curriculum. RESULTS/OUTCOME: This course provided participants with the skills to effectively utilize the Internet to locate high-quality medical information. EVALUATION METHOD: A course evaluation and impact survey were used to assess the effectiveness of the instruction. PMID- 14566370 TI - Woman to Woman: Community Health Information Project. AB - The Woman to Woman: Community Health Information Project facilitates information access at selected community-based women's health agencies in Houston, Texas. The community partners were selected based on objectives outlined in Healthy People 2010. A variety of institutions and disciplines contributed to the planning and implementation of the project. Internet-connected workstations were placed at each participating agency site for staff use. A comprehensive compendium of local health resources for women was developed and posted on a dedicated Website. In addition, training in accessing electronic health information resources was provided for staff at each participating site. Trainees provided evaluative data at the end of the training session as well as one month later so that project members could determine the need for additional training. Site visits and structured interviews were also conducted at each participating agency to monitor progress and further evaluate the program. A final focus group was conducted with representatives from each participating agency to assess the project's success. Evaluative findings indicate that academic and community-based organizations can collaborate effectively to increase women's access to electronic health information. PMID- 14566371 TI - HolisticKids.org--evolution of information resources in pediatric complementary and alternative medicine projects: from monographs to Web learning. AB - Use of complementary and alternative medicine (CAM) is growing in the United States. Children are a part of this trend, with adolescent self-care exceeding adult use. As a result, the necessity of educating pediatricians on CAM practices has become clear. This paper describes the Pediatric Integrative Medicine Education (PIME) project with a focus on the creation of HolisticKids.org, a Website designed to educate pediatric residents. HolisticKids.org also addresses the needs and interests of medical students, fellows, faculty, and community based care providers who work with children and CAM. An outstanding aspect of this effort is its interdisciplinary nature, involving physicians, pharmacists, dieticians, nurses, medical librarians, and Web specialists. Collaboration is mirrored on an institutional level, where the participating institutions include Children's Hospital Boston, Harvard Medical School, Boston Medical Center, the Massachusetts College of Pharmacy and Health Sciences, and Dana Farber Cancer Institute. Now in the third year of development, this Web-based education project demonstrates how integrative medical education can contribute to pediatric care for diverse families and complement cultural competency efforts. Ongoing efforts focus on adding content, developing evaluation techniques, and disseminating this resource to the health sciences community. PMID- 14566372 TI - Increasing access to Latin American social medicine resources: a preliminary report. AB - PURPOSE: This preliminary report describes the development and implementation of a project to improve access to literature in Latin American social medicine (LASM). METHODS: The University of New Mexico project team collaborated with participants from Argentina, Brazil, Chile, and Ecuador to identify approximately 400 articles and books in Latin American social medicine. Structured abstracts were prepared, translated into English, Spanish, and Portuguese, assigned Medical Subject Headings (MeSH), and loaded into a Web-based database for public searching. The project has initiated Web-based publication for two LASM journals. Evaluation included measures of use and content. RESULTS: The LASM Website (http://hsc.unm.edu/lasm) and database create access to formerly little-known literature that addresses problems relevant to current medicine and public health. This Website offers a unique resource for researchers, practitioners, and teachers who seek to understand the links between socioeconomic conditions and health. The project provides a model for collaboration between librarians and health care providers. Challenges included procurement of primary material; preparation of concise abstracts; working with trilingual translations of abstracts, metadata, and indexing; and the work processes of the multidisciplinary team. CONCLUSIONS: The literature of Latin American social medicine has become more readily available to researchers worldwide. The LASM project serves as a collaborative model for the creation of sustainable solutions for disseminating information that is difficult to access through traditional methods. PMID- 14566373 TI - The management of health library outreach services: evaluation and reflection on lessons learned on the VIVOS project. AB - PURPOSE: The aim of the VIVOS project was to develop and evaluate methodologies, i.e., sets of methods, for determining the value and impact of "virtual outreach" information services in the health sector in the UK. METHODS: Five different projects were recruited initially, with another two added later. Methods were largely qualitative, with over 130 interviews conducted among health professionals, complemented by postal questionnaire surveys. RESULTS: Identified factors that affect the successful roll-out and continued development of the projects included the need for help-desk type services to provide sustained support for new users to the services. CONCLUSIONS: Follow-up of the projects eighteen months after the end of the VIVOS project revealed that the long-term impacts for the participating library managers included the benefits of using evidence on service outcomes, enhanced recognition locally, and greater confidence in evaluation. PMID- 14566374 TI - Bibliometric study of grey literature in core veterinary medical journals. AB - OBJECTIVES: Grey literature has been perceived by many as belonging to the primary sources of information and has become an accepted method of nonconventional communication in the sciences and medicine. Since little is known about the use and nature of grey literature in veterinary medicine, a systematic study was done to analyze and characterize the bibliographic citations appearing in twelve core veterinary journals. METHODS: Citations from 2,159 articles published in twelve core veterinary journals in 2000 were analyzed to determine the portion of citations from grey literature. Those citations were further analyzed and categorized according to the type of publication. RESULTS: Citation analysis yielded 55,823 citations, of which 3,564 (6.38%) were considered to be grey literature. Four veterinary specialties, internal medicine, pathology, theriogenology, and microbiology, accounted for 70% of the total number of articles. Three small-animal clinical practice journals cited about 2.5-3% grey literature, less than half that of journals with basic research orientations, where results ranged from almost 6% to approximately 10% grey literature. Nearly 90% of the grey literature appeared as conferences, government publications, and corporate organization literature. CONCLUSIONS: The results corroborate other reported research that the incidence of grey literature is lower in medicine and biology than in some other fields, such as aeronautics and agriculture. As in other fields, use of the Internet and the Web has greatly expanded the communication process among veterinary professionals. The appearance of closed community email forums and specialized discussion groups within the veterinary profession is an example of what could become a new kind of grey literature. PMID- 14566375 TI - Emerging ethical issues in instructions to authors of high-impact biomedical journals. AB - Public interest in issues concerning the maintenance of high ethical standards in the conduct of scientific research and its publication has been increasing. Some of the developments in these issues as reflected in the publication of the medical literature are traced here. This paper attempts to determine whether public interest is reflected in the specific requirements for authors for manuscript preparation as stated in the "Instructions to Authors" for articles being prepared for submission to 124 "high-impact" journals. The instructions to authors of these journals were read on the Web for references to ethical standards or requirements. The ethical issues that the instructions most often covered were specifically related to the individual journal's publication requirements. The results suggest that while the editors and publishers of the biomedical literature are concerned with promoting and protecting the rights of the subjects of the experiments in the articles they publish, and while these concerns are not yet paramount, they are evolving and growing. PMID- 14566376 TI - Influence of information sources on the adoption of uterine fibroid embolization by interventional radiologists. AB - OBJECTIVES: The purpose of the research was to (1) understand the influence of information sources on the awareness and adoption of uterine fibroid embolization (UFE) by interventional radiologists in Michigan and (2) to decipher communication relations in the social network of interventional radiologists that were most conducive to the flow of information about UFE. METHODS: Diffusion of innovations theory and constructs in social network analysis formed the basis for the development of an interview guide. Thirty-two interventional radiologists in Michigan were interviewed over the phone. Chi-square statistics were employed to analyze the awareness and adoption of UFE. Factor analysis was applied to decipher important communication relations in the social network of interventional radiologists. RESULTS: Conferences were found to be an initial source of information, creating awareness among early adopters (P < 0.05), but other individuals were found to be influential sources in the adoption of UFE by later adopters (P < 0.05). Radiologists rarely browsed Websites for information. Work relations in everyday clinical practice were the communication relations most conducive to the flow of information about UFE. Preliminary qualitative data indicated that opinion leaders in the diffusion of UFE in Michigan were located in hospitals primarily dedicated to practice rather than in hospitals affiliated with universities. CONCLUSIONS: Journals are important information sources for creating awareness and stimulating adoption of innovation among both early and late adopters of new procedures in interventional radiology. Conferences, however, are significantly more important for creating early awareness, while interactions with colleagues is the most important factor in stimulating use of the innovation among later adopters. Among colleagues, opinion leaders in nonacademic hospitals may be more influential than individuals in the academic community. PMID- 14566377 TI - MEDLINE SDI services: how do they compare? AB - INTRODUCTION: Selective dissemination of information (SDI) services regularly alert users to new information on their chosen topics. This type of service can increase a user's ability to keep current and may have a positive impact on efficiency and productivity. Currently, there are many venues available where users can establish, store, and automatically run MEDLINE searches. PURPOSE: To describe, evaluate, and compare SDI services for MEDLINE. RESOURCES: The following SDI services were selected for this study: PubMed Cubby, BioMail, JADE, PubCrawler, OVID, and ScienceDirect. METHODOLOGY: Identical searches were established in four of the six selected SDI services and were run on a weekly basis over a period of two months. Eight search strategies were used in each system to test performance under various search conditions. The PubMed Cubby system was used as the baseline against which the other systems were compared. Other aspects were evaluated in all six services and include ease of use, frequency of results, ability to use MeSH, ability to access and edit existing search strategies, and ability to download to a bibliographic management program. RESULTS: Not all MEDLINE SDI services retrieve identical results, even when identical search strategies are used. This study also showed that the services vary in terms of features and functions offered. PMID- 14566378 TI - Information literacy skills of occupational therapy graduates: a survey of learning outcomes. AB - OBJECTIVES: The purpose of this study is to assess whether recent graduates of the Ohio State University's Occupational Therapy division are applying information-seeking skills they learned as undergraduates, and to seek their advice on ways to improve information-literacy instruction for current and future occupational therapy students. METHOD: A survey was sent to a sample of graduates from 1995-2000. The results were entered into an SPSS database, and descriptive and inferential results were calculated to determine the information-seeking patterns of these recent graduates. RESULTS: A majority of the occupational therapy graduates who responded to the survey prefer to use information resources that are readily available to them, such as advice from their colleagues or supervisors (79%) and the Internet (69%), rather than the evidence available in the journal literature. Twenty-six percent (26%) of the graduates have searched MEDLINE or CINAHL at least once since they graduated. Formal library instruction sessions were considered useful by 42% of the graduates, and 22% of the graduates found informal contacts with librarians to be useful. CONCLUSIONS: Librarians and occupational therapy faculty must intensify their efforts to convey the importance of applying research information to patient care and inform students of ways to access this information after they graduate. In addition to teaching searching skills for MEDLINE and CINAHL, they must provide instruction on how to assess the quality of information they find on the Internet. Other findings suggest that occupational therapy practitioners need access to information systems in the clinical setting that synthesize the research in a way that is readily applicable to patient-care issues. PMID- 14566379 TI - Testing the Database of International Rehabilitation Research: using rehabilitation researchers to determine the usability of a bibliographic database. AB - OBJECTIVES: This study tested the usability of the Database of International Rehabilitation Research, a bibliographic database developed by the Center for International Rehabilitation Research Information and Exchange (CIRRIE). METHODS: Potential users, i.e., rehabilitation researchers, were asked to participate in a usability study. Test questions were designed to represent common tasks performed in a bibliographic database. Participants were asked to think aloud during the test so that both their actions and comments could be recorded. RESULTS: This study identified common problems that participants had while searching the database and aspects of the database that needed improvement. CONCLUSIONS: Usability testing proved to be an effective method for evaluating database effectiveness and user satisfaction. The method used provided valuable information about how the database searchers approached their searches as well as how they performed them. PMID- 14566380 TI - Research methodology search filters: are they effective for locating research for evidence-based veterinary medicine in PubMed? AB - OBJECTIVE: The study examined the effectiveness of research methodology search filters developed by Haynes and colleagues and utilized by the Clinical Query feature of PubMed for locating literature for evidence-based veterinary medicine (EBVM). METHODS: A manual review of articles published in 6 commonly read veterinary journals was conducted. Articles were classified by format (original study, review, general article, conference report, decision analysis, and case report) and purpose category (etiology, prognosis, diagnosis, and treatment). Search strategies listed in PubMed's Clinical Query feature were then tested and compared to the manually reviewed data to calculate sensitivity, specificity, and precision. RESULTS: The author manually reviewed 914 articles to identify 702 original studies. Search #1 included terms determined to have the highest sensitivity and returned acceptable sensitivities over 75% for diagnosis and treatment. Search #2 included terms identified as providing the highest specificity and returned results with specificities over 75% for etiology, prognosis, and treatment. DISCUSSION: The low precision for each search prompts the question: Are research methodology search filters practical for locating literature for the practice of EBVM? A study examining terms related to appropriate research methodologies for advanced clinical veterinary research is necessary to develop filters designed to locate literature for EBVM. PMID- 14566381 TI - Teaching health informatics: designing a course for a new graduate informatics program. PMID- 14566382 TI - Probiotics: Isolated bacteria strain or mixtures of different strains? Two different approaches in the use of probiotics as therapeutics. AB - Probiotics are cultures of beneficial bacteria from the healthy gut microflora that improve the balance of the intestinal milieu by modifying the intestinal microflora and suppressing enhanced inflammatory responses. Probiotics are currently the subject of intense and widespread research as functional foods since they are known to induce health benefits, may be used as pharmaceutical preparations, and have achieved a "generally recognized as safe" (GRAS) status. Lactobacillus strains can also be genetically engineered for use in oral immunotherapeutic applications, such as vaccination and delivery of immunoregulatory substances. In the present review we evaluate the two different approaches to the therapeutic use of probiotics. We also focus on recent findings in the field of molecular biology and genetics of the intestinal immune response related to the microflora and intestinal ecology, in order to understand the mechanisms of action of probiotics and their present indications in gastrointestinal diseases. Finally, with a view to future perspectives we provide some examples of probiotics that are being assessed and have great potential in improving the health of animals and man. PMID- 14566383 TI - Role of 5-HT receptors in treatment of overactive bladder. AB - Normal bladder function requires coordinated detrusor relaxation and urethral sphincter contraction during the filling phase and the reverse during micturition. This is achieved by the integration of excitatory, inhibitory, and sensory nerve activity in control centers in the spinal cord, pons, and forebrain. It is possible that much of bladder pathology is related to disturbances in the vesical ganglia, sensory reflex loops, and central control of micturition. Experimental studies in animals have revealed that spinal reflex circuits involved in voiding function exhibit a dense serotonergic innervation, multiple 5-HT receptors, and sensitivity to 5-HT receptor agonists and antagonists, and 5-HT reuptake inhibitors. Activity in the serotonergic pathway generally enhances urine storage by facilitating the vesical sympathetic reflex pathway and inhibiting the parasympathetic voiding pathway. Thus, 5-HT receptor antagonists and reuptake inhibitors represent important targets for the development of new treatments of detrusor overactivity and urinary incontinence. Among the drugs modulating the bladder function through 5HT receptor, duloxetine, a potent and selective inhibitor of 5-HT and NE reuptake, is presently undergoing clinical trials for stress urinary incontinence after being shown to be effective in animal models of this condition. PMID- 14566385 TI - Present and future pharmacotherapy for osteoporosis. AB - Although neither calcium nor vitamin D has been shown to prevent osteoporosis in postmenopausal women alone, the combination does. Both calcium and vitamin D are commonly used in the treatment of osteoporosis. The estrogens and raloxifene both prevent bone loss in postmenopausal women, and the estrogens probably also decrease the risk of first fracture. There is good evidence that raloxifene prevents further fractures in postmenopausal women who have already had fractures and some evidence that estrogen does as well. Calcitonin increases bone mineral density in early postmenopausal women and men with idiopathic osteoporosis, and also reduces the risk of new fractures in osteoporotic women. The bisphosphonate alendronate prevents bone loss and reduces fractures in healthy and osteoporotic postmenopausal women, and in osteoporotic men. Risedronate is more potent and has fewer upper gastrointestinal side effects than alendronate, and reduces the incidence of fractures in osteoporotic women. Intermittent use of the potent bisphosphonate zoledronate also increases bone mineral density and may become an alternative in the prevention and treatment of osteoporosis. All of the agents discussed above prevent bone resorption, whereas teriparatide increases bone formation and is effective in the treatment of osteoporotic women and men. In the treatment of secondary osteoporosis associated with the use of glucocorticoids to treat inflammation or prevent rejection after transplantation, the bisphosphonates are effective. The agents that have undergone some clinical trialing as new or alternative drugs for the treatment of osteoporosis include tibolone, new SERMs, androgens, growth hormone, insulin-like growth factor-1 and stontium ranelate. The targets/drugs that are being developed to inhibit bone resorption include the OPG/ RANKL/RANK system, cathepsin K inhibitors, vitronectin receptor antagonists, estren, the interleukin-6 and gp130 system, cytokines and growth factors. New drugs/targets to promote bone formation include the commonly used lipid-lowering statins and the calcilytic release of PTH. PMID- 14566384 TI - Therapeutic significance of peroxisome proliferator-activated receptor modulators in diabetes. AB - Peroxisome proliferator-activated receptors (PPARs) are a group of three nuclear receptor isoforms, identified and encoded by different genes: PPARalpha, PPARdelta and PPARgamma. Each subtype of PPAR appears to be differently expressed in a tissue-specific manner due to its binding to a specific consensus DNA sequence of peroxisome proliferator response elements (PPREs). PPARalpha plays a significant role in the regulation of nutrient metabolism, including fatty acid oxidation, gluconeogenesis and amino acid metabolism. PPARdelta is expressed ubiquitously and has been found to be effective in controlling dyslipidemia and cardiovascular diseases, while PPARgamma isotype is mainly expressed in adipose tissue where it stimulates adipogenesis and lipogenesis. Thus PPARs have emerged as potential molecular targets for the design and synthesis of a different class of compounds, considering the conformation of receptors for the treatment of human metabolic disorders. This review concerns the therapeutic importance of PPARs in diabetes drug development. PMID- 14566386 TI - Substances which inhibit ice nucleation: a review. AB - There are a number of substances described in the published literature which inhibit ice nucleation. Certain bacterial strains, mostly found among the nonfluorescent pseudomonade species, release material into the growth medium which reduces the nucleation temperature of water droplets to below that of distilled water. Extracts from the seeds of food crops including apricot, peach and plum can reduce the nucleation temperature of water droplets and dispersions of silver iodide. Antifreeze glycoproteins can reduce the nucleation temperature of saline solutions. Antifreeze proteins can inhibit the activity of some biological ice nucleators but not others. Certain novel polymers have been shown to inhibit the nucleation activity of dispersions of silver iodide and ice nucleating bacteria. PMID- 14566387 TI - Antioxidative properties of lactoferrin from bovine colostrum before and after its lyophilization. AB - The effect of lactoferrin (LF) derived from native, frozen and lyophilized bovine colostrum on the intensity of free-radical processes in model systems has been investigated. It was shown that LF, not depending on the source of its obtaining, is an efficient iron chelator and decreases intensity of peroxidative processes. It was established, that antioxidative properties of LF from lyophilized colostrum have remained unchanged within 12 months of dry colostrum storage under proper conditions. PMID- 14566388 TI - Desiccation and freezing tolerance of embryonic axes from Citrus sinensis [L.] osb. pretreated with sucrose. AB - Embryonic axes of Citrus sinensis L. were successfully cryopreserved. While fully hydrated unfrozen axes germinated 100%, survival decreased as axes water content dropped, and total loss of viability was observed when the water content dropped to 0.04 and 0.10 mg H2O/mg dry mass, for axes without and with sucrose preculture, respectively. Fully hydrated axes did not survive exposure to liquid nitrogen. Highest seedling recovery (93-100%) for untreated axes was observed at 0.26 to 0.15 mg H2O/mg dry mass. Differential scanning calorimetry revealed the presence of broad melting peaks in fully hydrated embryonic axes. The size of the melting peak diminished as water was removed by desiccation. Minimum melting of water was observed at the point axes survived cryopreservation. Occurrence of a glass transition upon warming was not a condition for axes to survive liquid nitrogen exposure. In untreated axes, glucose, increased with desiccation to 0.2 mg H2O/mg dry mass, and decreased as the axes were desiccated to lower water contents. Fructose and sucrose levels did not increase when untreated samples were desiccated for the same periods of time. Raffinose and stachyose levels decreased as untreated and precultured embryonic axes were desiccated. In sucrose precultured axes, sucrose and fructose levels increased when they were dehydrated, reaching maximum levels at 0.2 mg H2O/mg dry mass. Tissue glucose did not change significantly with desiccation. Raffinose and stachyose levels dropped as precultured embryonic axes were dried. PMID- 14566389 TI - Effect of benzyladenine on recovery of cryopreserved shoot tips of grapevine and citrus cultured in vitro. AB - The effect of N6-benzyladenine (BA) on the recovery of cryopreserved shoot tips of the LN33 hybrid (Vitis L.) and Troyer citrange [Poncirus trifoliata (L.) Raf. x Citrus sinensis [L.] Osbeck.] cultured in vitro was examined. For the LN33 hybrid, the presence of BA in the recovery medium was essential for survival of control and cryopreserved shoot tips, although the BA concentration did not influence the survival percentage. BA at 5, 2, and 5 microM or higher induced callus formation in control, and shoot tips cryopreserved by vitrification, and by encapsulation-dehydration, respectively. While a BA concentration of 4 microM was found optimal for recovery of control shoot tips, 1 and 2-4 microM produced the best recovery of shoot tips cryopreserved by vitrification and encapsulation dehydration, respectively. A similar pattern of effect of BA on recovery was found for 'Troyer' citrange. Low survival of control and cryopreserved shoot tips was observed with a BA-free recovery medium. The addition of BA to the recovery medium significantly increased survival. The BA concentration that induced callus formation in shoot tips cryopreserved by encapsulation-vitrification was higher than that which induced it in those cryopreserved by encapsulation-dehydration. Recovery of control shoot tips was best with an addition of 6-10 microM BA to the medium. Optimal recovery of shoot tips cryopreserved by encapsulation vitrification and encapsulation-dehydration was achieved with 3-4 and 2 microM BA, respectively. Results from the present study suggest that an optimal BA concentration for recovery of control shoot tips may be different from that for cryopreserved shoot tips; furthermore, the optimal BA concentration for recovery of cryopreserved shoot tips may also differ among different cryogenic procedures. PMID- 14566390 TI - Cryopreservation of mango (Mangifera indica L.) embryogenic cultures. AB - Three techniques were compared for cryopreserving embryogenic masses (EMs) sampled from mango (Mangifera indica L.) cv. Zihua embryogenic cultures: (i) encapsulation/dehydration; (ii) pregrowth/dehydration; and (iii) vitrification. In all experiments, EMs were sampled from embryogenic cultures during their exponential growth phase and pretreated for 24 h on solid medium containing 0.5 M sucrose before freezing. No recovery was achieved after cryopreservation using the encapsulation/dehydration technique, whatever the moisture content (fresh weight basis) of EMs, which ranged from 78.3% without dehydration to 40.8% after 6 h dehydration. With the pregrowth plus dehydration technique, limited recovery (8.3%) was achieved after desiccation of EMs for 1 h, to 58.5% MC. Using the vitrification technique, recovery ranged from 94.3% after treatment of EMs with the PVS3 vitrification solution for 20 min (EM moisture content of 34.7%) to 10.9% after a 120 min treatment with the vitrification solution (EM moisture content of 26.0%). PMID- 14566391 TI - Genetic stability assessments of plantlets regenerated from cryopreserved in vitro cultured grape and kiwi shoot-tips using RAPD. AB - In vitro cultured shoot-tips of four grape (Vitis vinifera) cultivars and one kiwi (Actinidia deliciosa) cultivar were cryopreserved using the encapsulation dehydration method. Genomic DNA of plantlets regenerated directly from cryopreserved shoot-tips was extracted and analyzed using the RAPD (Random Amplified Polymorphic DNA) technique. The RAPD profiles obtained were highly reproducible and no differences were found between the DNA patterns obtained with plantlets regenerated from control and cryopreserved plantlets. The RAPD technique therefore appears to be a fast, simple and efficient method for evaluating genetic stability of cryopreserved material, which can be used rapidly after the completion of a freezing experiment and will efficiently complement other genetic stability evaluation methods. PMID- 14566392 TI - The effect of antifreeze proteins and poly(vinyl alcohol) on the nucleation of ice: a preliminary study. AB - Three substances have been tested for ice nucleation inhibition. These were an antifreeze protein AFP III from the fish Macrozoarces americanus, an antifreeze glycoprotein AFGP from the fish Dissostichus mawsoni, and an 80% hydrolysed poly(vinyl alcohol) with a molecular weight of 9 to 10 kD. A nucleation spectrometer was used to test nucleation inhibition at a range of concentrations against two types of ice nuclei: those present in tap water and a bacterial nucleator from Pseudomonas syringae. The PVA reduced the nucleation temperature of tap water and the bacterial dispersions at all the concentrations which were tested. The AFGP reduced the nucleation temperature of tap water but enhanced the nucleation activity of the bacterial nucleators. At low concentrations the AFP III reduced the nucleation temperature of both tap water and the bacterial nucleator. At high concentrations the AFP III enhanced the nucleation temperature of the bacterial nucleator and broadened the nucleation spectrum of the tap water to encompass the nucleation spread of the control. The possible mechanisms of nucleation suppression and enhancement are discussed. PMID- 14566393 TI - Rapid cold hardening in young hoppers of the migratory locust Locusta migratoria L. (Orthoptera: Acridiidae). AB - This paper describes a rapid cold hardening process for first instar hoppers of the migratory locust Locusta migratoria L. First instar hoppers of this species are often subjected to subzero temperatures or frosts in early April or May after their emergence from the soil. The mean supercooling point of hoppers is -13.0 +/ 1.4 degrees C; the fact that none could survive this temperature suggests they are freezing intolerant. When young hoppers were transferred directly from 30 degrees C to -7 degrees C for 2 h, there was only 35.8% survival. However, exposure to 0 degrees C for 2 h prior to transfer to -7 degrees C increased the apparent survival to 75%. A similar rapid cold hardening response can also be induced by gradual cooling at rates of between 0.05 and 0.1 degreess C min(-1). Rapid cold hardening also elevates the Ltime50 of first instar hoppers at -7 degrees C by approximately 3 fold, and reduces the lethal temperature from -10 degrees C to -12 degrees C. However, the protection from cold shock gained through rapid cold hardening was transient and easily lost within 2 h of hoppers being returned to 30 degrees C. The rapid cold hardening response is possibly advantageous to first instar hoppers that are often exposed to large temperature fluctuations in spring or early summer. PMID- 14566394 TI - Cardiac application of embryonic stem cells. AB - Cardiomyocytes deceased during myocardial infarction (MI) are replaced with non contractile scar tissue, which has a great chance to cause heart failure. Repair of dead or injured myocardium and improvement of cardiac function remain a serious challenge for the therapy of heart failure. Recently, stem cells have been transplanted in experimental settings to replace lost myocardium. This article summarizes the recent experimental findings on transplantation of embryonic stem cells (ESCs) and their derived cells in animals with myocardial injury and highlights the progresses in research of these particular cell types. Beneficial effects of cell transplantation with other cell types in injured hearts have been detailed in other reviews. ESCs are pluripotent cells derived from early mammalian embryos at the blasto-stage. These cells have the capacity for prolonged undifferentiated proliferation or differentiation into all of specialized somatic cell types of the body in culture, including cardiomyocytes. Because of the great ability of proliferation and differentiation to mature tissues, ESCs are a potential valuable resource for cell therapy targeting regeneration of functional myocardium in diseased hearts. In recent animal studies intramyocardial transplantation of ESCs or their differentiated cardiac like cells regenerated injured myocardium and improved heart function in infarcted animal models. In addition, intravenous infusion of ESCs significantly increased the survival rate and attenuated myocardial injury in viral myocarditic mice. Development and characterization of cardiomyocytes in vitro from human ESCs have been reported recently. However, many ethical, political, and scientific barriers have to be overcome before clinical utilization of human ESCs and their differentiated cells for treating end-stage cardiac diseases. PMID- 14566395 TI - Intrarenal artery injection of capsaicin activates spontaneous activity of renal afferent nerve fibers. AB - The effects of intrarenal artery injection of capsaicin on multi- and single-unit spontaneous discharges of renal afferent nerve fibers were investigated in anesthetized rabbits. The results obtained are as follows: (1) intrarenal artery injection of capsaicin (20, 40, and 60 nmol/kg) increased the renal afferent nerve activity (ARNA) in a dose-dependent manner with unchanged arterial pressure; (2) pretreatment with ruthenium red (40 mmol/kg), a capsaicin receptor antagonist, completely abolished the effect of capsaicin; and (3) pretreatment with a nitric oxide synthase inhibitor L-NAME (N(6)-nitro-L-arginine methylester, 0.1 mmol/kg), significantly enhanced the ARNA response to capsaicin. The results suggest that intrarenal artery injection of capsaicin can activate ARNA via capsaicin receptors in anesthetized rabbits and that nitric oxide may be involved in regulating the activity of renal sensory nerve fibers as an inhibitory neurotransmitter. PMID- 14566396 TI - Electrophysiological effects of capsaicin on guinea pig papillary muscles. AB - The cardiac electrophysiological effects of capsaicin (CAP) were examined in guinea pig papillary muscles using intracellular microelectrode technique. The results obtained are as follows: (1) the duration of action potential (APD) in normal papillary muscles was decreased by CAP (30, 60, 120 micromol/L) in a concentration-dependent manner; (2) in partially depolarized papillary muscles, 60 micromol/L CAP not only reduced APD, but also decreased the amplitude of action potential, overshoot and maximal velocity of phase 0 depolarization; (3) pretreatment with L-type Ca(2+) channel agonist Bay K8644 (0.5 micromol/L) could completely block the effects of CAP (60 micromol/L); (4) pretreatment with ruthenium red (20 micromol/L), a vanilloid receptor (VR) blocker, did not affect the actions of capsaicin on papillary muscles. All these results suggest that the effects of CAP on papillary muscles are likely due to a decrease in calcium influx which is not mediated by VR. PMID- 14566397 TI - Effects of intravenous Injections Paederiae and Stauntonia on spontaneous pain, hyperalgesia and inflammation induced by cutaneous chemical tissue injury in the rat. AB - To study whether commercial traditional Chinese medicinal preparations Injection Paederiae (IP) or Injection Stauntonia (IS) has anti-nociceptive and/or anti inflammatory effects, we used two persistent pain models (bee venom and formalin test) to evaluate the systemic effects of IP or IS on the chemical tissue injury induced persistent spontaneous pain-related responses (PSPR), primary thermal/mechanical hyperalgesia and inflammation in conscious rats. Injection of bee venom (BV, 0.1 mg, 50 microl) into the plantar surface of one hind paw resulted in not only a 1-h monophasic PSPR such as flinching reflex in the injected paw and a subsequent period of 3-4 days primary heat and mechanical hyperalgesia, but also a marked sign of inflammation, including redness and swelling of the plantar surface in the injected paw. Intraplantar injection of formalin produced two phases of PSPR as reported previously. Systemic pre treatment with three doses of IP (0.32, 1.6 and 9.0 ml/kg, 500%) or IS (0.32, 1.6 and 9.0 ml/kg, 250%) produced a dose-dependent suppression of the BV- or formalin induced flinching reflex of 1 h time course as compared with the saline control group. Post-treatment with IP or IS 5 min after BV injection also produced a significant suppression of the flinching reflex in both BV test and formalin test respectively, as compared with the control group. However, neither pre- nor post treatment with IP or IS produced any significantly suppressive effect on the BV induced primary heat and mechanical hyperalgesia and inflammation. The analgesia produced by IP or IS was not mediated by the endogenous opioid receptors since naloxone, a non-selective opioid receptor antagonist, had no reversal effect on the IP and IS-produced analgesia in the BV-induced PSPR. Our present results suggest that IP or IS might prevent and relieve clinical persistent spontaneous pain, but without any anti-nociceptive and anti-inflammatory effects on the primary heat hyperalgesia, mechanical hyperalgesia, as well as inflammatory responses. The BV test might be a useful model of pain to evaluate and screen anti-nociceptive and anti-inflammatory effects of certain compounds of the Chinese medicinal herbs on the pathological origins of pain. PMID- 14566398 TI - Synergistic regulation of the acute phase protein SIP24/24p3 by glucocorticoid and pro-inflammatory cytokines. AB - SIP24/24p3 is a secreted murine acute phase protein which has been speculated to play an anti-inflammatory role in vivo. Recently SIP24/24p3 has been found to be able to specifically induce apoptosis in leukocytes. By using (35)S metabolic labeling method, we studied the regulation of SIP24/24p3 by glucocorticoid and pro-inflammatory cytokines IL-6 and TNF-alpha in cultured Balb/c 3T3 and BNL cells. The following results were observed: (1) dexamethasone induced the expression of SIP24/24p3 in both Balb/c 3T3 and BNL cells, the induction was more significant in BNL cells; (2) dexamethasone and IL-6 synergistically induced the expression of SIP24/24p3 in both Balb/c 3T3 and BNL cells; (3) in Balb/c 3T3 cells dexamethasone and TNF-alpha acted synergistically to induce the expression of SIP24/24p3, whereas in BNL cells dexamethasone and TNF-alpha induced the expression of SIP24/24p3 in an additive manner; (4) dexamethasone and IL-6/TNF alpha acted synergistically in Balb/c 3T3 cells and additively in BNL cells to induce the expression of SIP24/24p3. The inducibility of SIP24/24p3 by multiple factors will help to explain its highly specific expression in vivo. The difference in the expression patterns of SIP24/24p3 in different cell types is also suggestive to its expression and regulation in hepatic and extrahepatic tissues. Finally, the fact that SIP24/24p3 protein can be induced by both pro inflammatory as well as anti-inflammatory factors is indicative of the important role of SIP24/24p3 in the entire acute phase response process. PMID- 14566399 TI - BmK I, an alpha-like scorpion neurotoxin, specifically modulates isolated rat cardiac mechanical and electrical activity. AB - In this study, cardiotonic and cardiotoxic effects of Buthus martensi Karsch (BmK) I, a modulator of voltage-gated sodium channels, were investigated on the isolated rat hearts. The results showed that BmK I evoked complex effects characterized by a change in both cardiac mechanical and electrical activity. Langendorff perfusion showed that: (1) maximal left ventricular developed pressure (LVDP(max)) and dp/dt(max) were markedly increased by BmK I (0.5-10 micromol/L) in a dose-dependent manner (n=6, P<0.05), positive chronotropic effects were also induced by BmK I (n=6, P<0.05); (2) negative inotropic action and bradycardia could be elicited at a larger dose of BmK I (20 micromol/L); (3) the coronary flow varied inversely with the positive inotropic effects, coronary flow reduced during positive inotropic effects from 14.5 to 8.6 ml/min after administration of 500 nmol/L BmK I (n=6, P<0.05). In addition, tachycardia and complex cardiac arrhythmias were induced by BmK I (0.5-10 micromol/L). The modulating of BmK I on the heart mechanical, electrical activity could be partially recovered after washing. As propranolol was applied to block the release of catecholamines before administration of BmK I, suggesting that the changes in cardiac mechanical and electrical activity induced by BmK I might not due to catecholamine release from the nerve terminal and subsequent stimulation of the beta-adrenoceptor but attributable to the modulation of BmK I on cardiac voltage-gated sodium channels. PMID- 14566400 TI - Action of NO and TNF-alpha release of rats with cadmium loading in malfunctiion of multiple system organ. AB - Thirty-six healthy Sprague-Dawley male rats were used and divided randomly into a control group (group C), a medium-dose cadmium loading group (group M) and a high dose cadmium loading group (group H). Cadmium chloride was diluted with saline to contain 0.4 mg/ml of autoclaved cadmium solution. Groups M and H were injected in the abdominal cavity with 0.5 and 1.0 mg of cadmium per kg body weight respectively, and group C with saline of the same dosage as in group H over 7 d. Six rats of each group were killed on the 4th day and 7th day after cadmium loading, respectively, and blood, testis, liver and heart were collected. Cadmium content, changes in nitric oxide and tumor necrosis factor-alpha were studied in blood and the tissues of testis, liver and heart. Results showed that during the entire experimental period, body weight in groups M and H decreased significantly as compared with that in group C; cadmium concentration increased significantly in testis, heart and liver of groups M and H, and rose with increased dosage and time of cadmium loading; there was no obvious difference in plasma nitric oxide between groups M and C, though nitric oxide was higher in group M than in group C. Nitric oxide in group H was significantly superior to that in group C. Plasma tumor necrosis factor-alpha was markedly higher in groups M and H than in group C. Nitric oxide contents in the rat s testis, liver and heart homogenates with cadmium loading were higher than those in group C or significantly superior to group C. The same changes in tumor necrosis factor-alpha in the tissue homogenates of the testis and heart were found, but no obvious difference in tumor necrosis factor-alpha in the liver between the three groups was observed. It is suggested that the massive release of nitric oxide and tumor necrosis factor-alpha induced by cadmium loading may play an important role in the induction of malfunction of multiple systems or organs in rats. PMID- 14566401 TI - [Accurate establishment of the retinotopic topography of area 17 in cats by intrinsic signal optical imaging]. AB - The retinotopic topography of area 17 in cats was measured by optical imaging based on intrinsic signals. When stimulated with two neighboring gratings oriented orthogonally each other, which were positioned respectively in the upper and lower visual fields, one piece of cortex that had the retinal projection corresponding to the area around the border of the two stimulus gratings became blurred in the resultant function orientation map, because the neurons in this site received excitatory signals from both the horizontal and the vertical gratings via indirect ways. This functional map of the same cortex was compared with that elicited only by a horizontal or vertical grating stimulation in the whole visual field. Accordingly, the accurate position of the retinotopic eccentricity of the cortex in visual field can be demarcated by calculating the cross correlation coefficient of the two functional maps. Furthermore, compared with the electrophysiological measure of receptive fields of single cortical neurons, the retinotopic eccentricities revealed by optical imaging were identical. This experiment provides a fast and relatively accurate method to calculate the retinotopic eccentricities in a large cortical area of the visual cortex. PMID- 14566402 TI - Changes in diurnal rhythms of free cortisol secretion during different phases of menstrual cycle. AB - The effect of the menstrual cycle on the diurnal cortisol rhythm was investigated in 15 normally cyclic healthy women during reproductive life. Salivary cortisol was measured by radioimmunoassay in samples collected every 2 h for 24 h during the four phases of the menstrual cycle: menstrual phase, late follicular/peri ovulation phase, early to mid luteal phase and late luteal phase, respectively. Distinct diurnal rhythms of free cortisol were found throughout the menstrual cycle by using a nonlinear periodic regression model. The model was characterized by an asymmetrically peaked diurnal cycle and ultradian harmonics. There was a trend to higher troughs and significantly shorter peak-width in phase II and phase IV compared to phase I. The ultradian amplitude in phase IV was significantly lower compared with phase I and showed a trend of decrease compared with phase II. The results suggest that the daily cortisol secretion is modulated by the phase of the menstrual cycle. PMID- 14566403 TI - Effects of endothelial cell growth states on the proliferation and migration of vascular smooth muscle cells in vitro. AB - Endothelial injury, smooth muscle cells (SMCs) proliferation and migration are the same common pathophysiological processes of many cardiovascular diseases, such as atherosclerosis, hypertension, diabetes and restenosis. It is important to determine the functional interactions between endothelial cells (ECs) and SMCs under pathologic conditions. This work was to study the effects of ECs growth states on the proliferation and migration of vascular SMCs in cell coculture system. (3)H-TdR incorporation and flow cytometry were used to determine the effects of ECs growth states on the proliferation of SMCs. The number of migrating SMCs was counted. RT-PCR was used to analyze the expression of alpha-SM actin mRNA. The results showed that (3)H-TdR incorporation decreased significantly from 14,900+/-1035 cpm/well in the control group to 8,575+/-749 cpm/well in the confluent ECs group (n=6, P<0.01), and increased to 27,268+/-2310 cpm/well in the proliferative ECs group ( n=6, P<0.01). The transition of SMCs from G(0)/G(1) phase to G(2)/M and S phases was blocked in the confluent ECs group but promoted in the proliferative ECs group. Compared with the control group, the number of migrating cells was about 4 times higher in the proliferative ECs group (n=6, P<0.01), while it in the confluent ECs group was only the half of the number of the control(n=6, P<0.05). The expression of alpha SM-actin mRNA was increased significantly in the confluent ECs group(2.3+/-0.11 vs 1.4+/-0.12, P<0.05), but decreased significantly in the proliferative ECs group(0.92+/-0.08 vs 1.4+/-0.12, P<0.05). The results suggest that the biologic features of SMCs are influenced by ECs growth states. The proliferative ECs promote SMCs proliferation, migration and downregulate alpha-SM-actin mRNA expression significantly. PMID- 14566404 TI - [Nitric oxide is involved in the modulation of central respiratory rhythm]. AB - This experiment was expected to test whether nitric oxide (NO) exerted significant effect on the central respiratory rhythm. Experiments were performed on in vitro brainstem slice preparations from neonatal rats. These preparations include the medial region of the nucleus retrofacialis (mNRF); a part of pre Botinger complex, ventral respiratory group (VRG) and dorsal respiratory group (DRG). Respiratory-related burst activities were recorded from hypoglossal nerve rootlets before and during superfusion of the slice preparation with L-Arginine (L-Arg), sodium nitroprusside (SNP) or 7-nitro indazole (7-NI, an inhibitor of NO synthase). After perfusion with L-Arg and SNP, there was no significant change in respiratory rhythmical discharge activity (RRDA), but 7-NI decreased the integral amplitude of burst and inspiratory time. These results indicate that NO may take part in the inspiratory off-switching mechanism and that it also modulates the amplitude of respiratory-related bursts. PMID- 14566405 TI - [Effects of D1 and D2 dopamine receptor agonists and antagonists on cerebral ischemia/reperfusion injury]. AB - Gerbil forebrain ischemia/reperfusion(I/R) injury model was used to study the effects of D(1) and D(2) receptor agonists and antagonists on neuronal apoptosis of hippocampal CA1 area. All animals were tested for habituation deficits in an open field test on the 1st, 3rd and 7th days after reperfusion. The animals were then killed, and brains underwent paraffin embedding for hematoxylin-eosin staining, in situ terminal deoxynucleotidyl transferase-mediated deoxy-UTP nick end labeling (TUNEL) staining and immunohistochemistry (bax, bcl-2). The result of open field test showed that the I/R group was significantly impaired (higher activity scores) when compared with the control group. Pretreatment with pergolide significantly reduced this habituation impairment. Forebrain ischemia for 5 min resulted in extensive CA1 apoptosis on the 3rd and 7th days after I/R injury. About 95% neurons in hippocampal CA1 area entered apoptosis and only 2% 7% pyramidal neurons stayed alive due to an inhibition of bcl-2 expression and an increase in bax expression. Pretreatment of pergolide attenuated neuronal damage caused by transient ischemia. Infusion of pergolide could induce the expression of bcl-2 and reduce the expression of bax. Pretreatment with SKF38393, SCH23390 and spiperone had no effects on these changes in this transient I/R injury model. All these results indicate that pergolide plays an important role in the protection of hippocampal neurons from apotosis through upregulating the expression of bcl-2 protein and reducing the expression of bax protein. PMID- 14566406 TI - [Increased DMT1 expression and iron content in MPTP-treated C57BL/6 mice]. AB - Iron plays a key role in Parkinson s disease (PD). To illustrate the mechanism underlying the increase of iron in substantia nigra (SN) in PD, changes of the expression of divalent metal transporter 1 (DMT1) and iron content were examined in SN in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) treated mice using immunohistochemistry and histochemistry respectively. Following MPTP treatment for 3 d, elevated iron staining was found in SN. A further increase in iron content was observed after 7 d. In these lesioned animals, tyrosine hydroxylase immunoreactive DA neurons exhibited a decrease in number and morphological changes as well. There were two isoforms of DMT1 expressed in SN of mice. After MPTP treatment, the expression of DMT1 without IRE form increased in either group, whereas DMT1 with IRE form increased only after 7 d of MPTP treatment. These observations suggest that DMT1 is possibly involved in the process of iron accumulation in SN of MPTP-treated mice, which might be responsible for the subsequent death of DA neurons. PMID- 14566407 TI - [Microinjection of L-NAME into dorsal raphe nucleus inhibits nociceptive response in sigmoid pain model of rats]. AB - By means of Fos immunocytochemistry, nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry and microinjection methods, the role of nitric oxide synthase (NOS) of dorsal raphe (DR) neurons in the modulation of rats sigmoid pain was studied. The results showed: (1) Rats exhibited aversive behavioral responses related to visceral pain after injecting formalin into the sigmoid wall. NOS neurons in DR were up-regulated, in addition, about 8% of NOS labeled neurons were Fos positive. By contrast, there were no Fos/NOS double labeled neurons in the control group. (2) Formalin-induced sigmoid pain scores and the expression of Fos in the spinal cord at S1 segment were decreased after microinjecting L-NAME into the DR. These findings suggest that NOS neurons are involved in the modulation of formalin-induced sigmoid pain and that NO may play an important role in the transmission of visceral nociceptive message in the midbrain. PMID- 14566408 TI - [Expression and assessment of double genes of tyrosine hydroxylase gene and aromatic L-amino acid decarboxylase gene in vitro]. AB - The characteristic pathological changes of Parkinson s disease (PD) include a severe loss of dopamine neurons in the substantia nigra and a severe decrease in dopamine in the striatum. Since the expression of tyrosine hydroxylase (TH) and aromatic L-amino acid decarboxylase (AADC) in the biosynthetic pathway for dopamine are low, a promising approach to the gene therapy of PD is to augment the gene expression of the enzymes in the biosynthetic pathway for dopamine. In the present study, human TH and AADC genes were reconstructed into retrovirous vectors pLHCX and pLNCX(2) respectively. Then pLHCX/TH and pLNCX(2)/AADC were transfected into packaging cell line PA317 with liposome. PA317/TH and PA317/AADC were selected by different antibiotics. Gene expression was examined by methods of immunohistochemistry and in situ hybridization. The catalytic activity of two cloned gene enzymes was assessed in vitro by HPLC-EC. Immunocytochemical staining showed that TH and AADC were expressed efficiently in vitro. Both TH and AADC mRNA were transcripted in PA317 cell lines by using in situ hybridazation. HPLC EC experiments revealed that the transfected cells produced a significantly higher level of dopamine and L-dopa than the untransfected cells. The two genetically modified cells could improve the production of L-dopa and dopamine in response to suitable substrate. The present results suggest that not only recombinant TH and AADC genes are successfully expressed in vitro, but also the enzymes have respective functional activities. These results have set up a way for in vivo gene therapy of PD with TH and AADC genes. PMID- 14566409 TI - [Regulation of estrogen and phytoestrogen on the dopaminergic systems of amygdala in rats]. AB - In vivo fast cyclic voltammetry (FCV) was used to investigate dopamine (DA) release from amygdala (Amy) of female rats in different phases of estrus cycle, ovaricectomized (OVX) rats and male rats. Tyrosine hydroxylase (TH) immunohistochemistry was employed to measure the numbers of immunoreactive neurons in ventral tegmental area (VTA) of midbrain in the rats. We also observed the effect of intracerebroventricular injection of phytoestrogen-soy isoflavones on DA release from the Amy. The results are as follows: DA release from the Amy of proestrus female rats was apparently higher than that in estrus, metaestrus, diestrus female rats and OVX rats. Amy DA release and the numbers of the TH immunoreactive neurons in VTA showed a significant sex difference. DA release from Amy of female and OVX rats increased significantly within 5 min after i.c.v injection of soy isoflavones, which elicited no effects in male rats. The above mentioned results suggest that endogenous estrogen may play an important role in regulating the activity of DA neurons in mid-limbic systems, and that soy isoflavones exert an estrogen-like effect on the dopaminergic systems in the Amy. PMID- 14566410 TI - [Role of calcineurin in the progression of cardiac hypertrophy in renovascular hypertensive rats]. AB - The present study was to investigate the mRNA, protein expression and the activity of calcineurin in the hypertrophic heart, and to determine the effect of calcineurin inhibitor--cyclosporine A (CsA) on the regression of cardiac hypertrophy in renovascular hypertensive rats. Renovascular hypertension was induced by two kidney-one clip methods. Two months after the operation, cardiac hypertrophy was determined by histological analysis performed in some rats (2K1C 2M), then the rats were subdivided into 2 groups: (1) 3-month old two kidney-one clip group (2K1C-3M) with rats receiving 0.9% NaCl per day for one month, and (2) CsA-treated group with rats treated with CsA for one month. Sham-operated rats were used as control. The ratio of the left ventricular weight to tibial length (LVW/TL), the area of cardiac myocyte, mRNA and protein expression and the activity of calcineurin were determined. Both the LVW/TL and the cardiomyocyte area were significantly larger in 2K1C-2M and 2K1C-3M rats than in age-matched sham-operated rats. Treatment with CsA significantly attenuated the increase in the LVW/TL as well as the cardiomyocyte area. The mRNA, protein expression and the activity of calcineurin were significantly higher in 2K1C-2M and 2K1C-3M rats than those in the age-matched sham-operated rats, while the elevation of mRNA, protein expression and activity of calcineurin were significantly suppressed in the CsA-treated rats. In conclusion, calcineurin plays a role in the progression of cardiac hypertrophy in renovascular hypertensive rats. The inhibition of calcineurin can reverse cardiac hypertrophy. PMID- 14566411 TI - [Modulation of extracellular calcium on miniature inhibitory postsynaptic currents of Xenopus' optic tectal neurons]. AB - Experiments were performed to study the relationship between presynaptic calcium and miniature inhibitory postsynaptic currents (mIPSCs) using voltage-clamp technique with whole cell mode in Xenopus optic tectal slices. The results are as follows: The frequency of mIPSCs decreased from 1.91+/-0.59 Hz to 0.34+/-0.09 Hz in calcium-free solution (paired t test, P=0.019, n=8), the value of mIPSCs frequency being (25.5+/-4.4)% of control. In order to rule out the effect of remaining calcium in perfusing solution we further applied calcium-free solution containing egtazic acid, ethyleneglycol-bis (beta-aminoethylether)-N,N tetraacetic acid (EGTA) (200 nmol/L-2 mmol/L). The mIPSCs frequency changed from 1.57+/-0.57 in control to 0.89+/-0.41 Hz in calcium-free solution containing EGTA (paired t test, P=0.002, n=12), decreasing to (40.0+/-5.9)% of control. There is no statistical difference in the results between Ca(2+)-free perfusion and Ca(2+) free solution containing EGTA (paired t test, P=0.74, n=9). When cadmium chloride (CdCl2, 100 micromol/L), a non-specific Ca(2+) channel blocker, was applied to the bath solution, the mIPSCs frequency decreased from 1.15+/-0.34 Hz in control to 0.25+/-0.09 Hz in CdCl2-containing solution (paired t test, P=0.008, n=11), reaching (29.25+/-6.1)% of control. However, the amplitude did not change much. An endoplasmic reticulum pump inhibitor thapsigargin increased the mIPSCs frequency from 0.93+/-0.19 Hz to 1.58+/-0.28 Hz (paired t test, P=0.002, n=11). The value in the latter is (214.6 +/-49.1)% of that in the former. In order to exclude the remaining calcium from the bathing solution, the mIPSCs frequency was first recorded from calcium-free solution as control (0.41+/-0.08 Hz) and then from calcium-free containing TG solution (8-16 micromol/L)(0.71+/-0.15 Hz)(paired t test, P=0.026, n=5), increasing to (175.0+/-14.6)% of control. The endoplasmic reticulum Ca(2+) store RyR agonist (ryanodine, 10-100 nmol/L) enhanced mIPSCs frequency from 1.18+/-0.40 Hz to 1.80+/-0.44 Hz with increment of (261.8+/-89.5)% (paired t test, P=0.004, n=6). However, the endoplasmic reticulum RyR antagonist (procaine, 2 mmol/L) could inhibit mIPSCs from 1.26+/-0.35 Hz to 0.43+/-0.15 Hz (paired t test, P=0.027, n=6). U73122 (40 micromol/L), a phosphalipase C inhibitor, decreased also mIPSCs frequency from 2.01+/-0.58 Hz in control to 0.92+/-0.40 Hz in U73122-containing solution (paired t test, P=0.002, n=10). Caffeine (10 mmol/L) markedly diminished mIPSCs frequency from 3.22+/-0.64 Hz to 0.15+/-0.30 Hz (paired t test, P=0.003, n=7), which is (4.6+/-2.9)% compared to control. Furthermore, in some cases the caffeine could abolish mIPSCs. Taken together, our results demonstrated that cytosolic calcium might be important for mediating the generation of mIPSCs. The cytosolic calcium could be increased by calcium influx through membrane calcium channel on presynaptic membrane, and/or by calcium released through RyR and IP(3)R in presynaptical internal store. The increased cytosolic Ca(2+) both from external solution or internal Ca(2+) stores might increase the transmitter vesicles at the presynaptic terminal, which in turn results in the increase of the mIPSCs frequency in the postsynaptic neurons. PMID- 14566412 TI - [Time domain and power spectrum of wide frequency band electrocardiogram in pigeons]. AB - The wide frequency band ECG (WFB-ECG) was recorded in 33 (anesthetized) normal pigeons by the microprocessor ECG system (made in Nanjing University) with a wide frequency response (0-1000 Hz), a high-speed sweep (up to 1401 mm/s) and a high sensitivity (up to 28 mm/mV). The recording methods for limb leads in the pigeon were the same as those in man, except that the needle electrodes (made by No.5 needles) were subcutaneously inserted in the bases of the wings and in the legs. We studied the features of time domain and power spectrum of pigeons WFB-ECG. It presents P, R, S and T waves, but no Q wave, basically similar to the results from Aves described by Sturkie. But there are still many characters that were not be recorded on the conventional ECG: (1) the main QRS complex is inverted and forms the type of rS or rSr , no Q wave in leads II, III, aVF, and the S-T segment is absent, which is different from that of humans. The T wave is upright in leads II, III, and aVF (except one), in agreement with that of man. But in lead aVR, the main QRS complex is upright and forms the type of Rs, and the T wave is inverted without any exception. There is a large notch on the upstroke of S wave without any exception. The amplitude of the notch is 0.413+/-0.133 mV and the duration is 9.733+/-1.291 ms in lead II. (2) The ratio of duration of P wave to P-R segment is about 0.8, lower than that of humans (1.0-1.6), but higher than that of mice (0.4). (3) The low frequency signals (0-80 Hz) are prominent. The relative power content of high frequency range of QRS in lead II is: 100-1000 Hz: (10.181+/-7.443)%; 80-300 Hz: (15.418+/-10.579)%. (4)The QRS vector loop in the frontal plane lies between -90 degrees and -180 degrees. The electrical axis of QRS complex averages -118 +/-10 (ranges from -96 degrees to -136 degrees). The reason that position of vector loop and the direction of main wave of QRS in the pigeon are different from human s and rodent s is probably that the Purkinje fibers cross the whole ventricular wall and terminate in the subepicardium in Aves including pigeons . After the impulses coming from the sinoatrial node reach the ventricular muscles, the subepicardium is depolarized before the endocardium. However in human s and rodent s, the Purkinje fibres only reach one-forth to one second of the whole thickness from the endocardium to the epicardium, the subendocardium is depolarized before the subepicardium. PMID- 14566413 TI - [Application of combination of capillary electrophoresis with laser-induced fluorescence: measurement of glutamate and arginine in PAG microdialytes of conscions morphine-withdrawal rats]. AB - Microdialysis technique in free-moving animals can be used to monitor continuously the changes of many extracellular neurotransmitters in certain brain areas and study the relationship between neurotransmitter and functions. Using detection of capillary electrophoresis combined with laser-induced fluorescence (CE-LIF) further improves the above-mentioned technique. In the present study, fluorescein isothiocyanate (FITC) was used to derivatizate amino acid in very low concentration. We found that increasing derivatization temperature could shorten derivatization time and that the derivatizative efficiency was not different from that when experiment was performed under the traditional derivatization condition (room temperature for 16 h). We also got an optimized condition of amino acid derivatization with FITC at 30 degrees C water bath for 5 h. Using the optimized condition of amino acid derivatization, we investigated the changes in L-arginine (L-Arg) and L-glutamate (L-Glu) concentration in periaqueductal gray matter (PAG) microdialytes of free-moving morphine-withdrawal rats. The results indicated that there was no significant difference in the concentration of L-Arg and L-Glu in PAG between non-dependent and dependent rats. The concentration of L-Arg and L Glu in PAG increased by 63% and 105%, respectively, in the first 10 min after naloxone-precipitated withdrawal and then declined gradually. These changes were in correspondence with the scores of morphine withdrawal symptom. PMID- 14566414 TI - Lissencephaly with agenesis of corpus callosum and rudimentary dysplastic cerebellum: a subtype of lissencephaly with cerebellar hypoplasia. AB - Lissencephaly with agenesis of the corpus callosum and rudimentary dysplastic cerebellum may represent a subset of lissencephaly with cerebellar hypoplasia (LCH) of unknown etiology, one that is distinct from other types of LCH. We present a detailed neuropathological description of an autopsy brain from a 7-day old neonate born at 38-gestational weeks, presenting with this malformation. The brain was severely hydrocephalic and totally agyric. The corpus callosum was absent and deep gray matter structures indistinct. A rudimentary dysplastic cerebellum, dysplastic olivary nuclei and nearly complete absence of corticospinal tracts were also noted. Microscopic examination revealed various types of dysplastic and malformative features throughout the brain in addition to the classic four-layered neocortical structure characteristic of type I lissencephaly. Unique features in the present case were (1) bilateral periventricular undulating cortical ribbon-like structures mimicking fused gyri and sulci, associated with aberrant reelin expression, (2) large dysplastic neocortical neurons positive for phosphorylated neurofilament, calbindin-D28K, tuberin, hamartin, doublecortin, LIS1, reelin and Dab1, (3) derangement of radial glial fibers, and (4) disorganized cerebellar cortex and heterotopic gray matter composed exclusively of granule cells in the cerebellar deep white matter. The clinicopathological features in the present case are suggestive of a distinct category of lissencephaly with cerebellar involvement. We suggest a possible classification of this unique case among the LCH syndromes. PMID- 14566415 TI - Absence of allelic loss in cytomegalic neurons of cortical tuber in the Eker rat model of tuberous sclerosis. AB - The Eker rat is an animal model of tuberous sclerosis caused by a mutation in the Tsc2 gene encoding a tumor suppressor protein, tuberin. According to Knudson's two-hit theory, renal carcinomas and other tumors develop in various organs. Although the incidence of brain lesions is lower in the Eker rat than in human tuberous sclerosis, a cortical tuber was recently found in the cerebrum of an Eker carrier. In this study, we examined whether neuronal cytomegaly in the Eker rat tuber is caused by deletion of the normal Tsc2 allele and resultant loss of tuberin, as is the case with the majority of renal carcinomas. A combination of laser capture microdissection and semi-nested polymerase chain reaction demonstrated the presence of the wild-type Tsc2 allele in the cytomegalic neurons isolated individually. Immunohistochemistry also detected positive tuberin immunoreactivity in many of these giant neurons. These findings were in sharp contrast to those of renal carcinoma cells deriving from allelic loss. Our results provide evidence that many if not all cytomegalic neurons of a cortical tuber occur in the absence of allelic loss. PMID- 14566416 TI - Interstitial cells of Cajal are normally distributed in both ganglionated and aganglionic bowel in Hirschsprung's disease. AB - Surgery for Hirschsprung's disease is often complicated by post-operative bowel motility disorders. The impact of intestinal neural histology on the surgical outcome has been previously studied, but no information is available concerning the influence of the distribution of interstitial cells of Cajal (ICC) on these complications. These cells are considered to be pacemakers in the gastrointestinal tract. The aim of this study was to assess the distribution of ICC in the proximal segment of resected bowel in Hirschsprung's disease and confront these results with the clinical outcome. Using immunohistochemistry for light microscopy, we compared the pattern of distribution of ICC in the proximal segment of resected bowel in Hirschsprung's disease with that in normal colon. We correlated these results with the corresponding neural intestinal histology determined by CD56 and the protein gene product 9.5 immunohistochemistry. The distribution of ICC in the proximal segment of resected bowel is identical to that of normal colon, regardless of normal or abnormal colon innervation. ICC distribution does not seem to contribute to post-operative bowel motility disorders in patients operated for Hirschsprung's disease PMID- 14566417 TI - A new technique for esophagoplasty in relatively long esophageal strictures. AB - A 3-year-old girl with severe dysphagia secondary to caustic stricture of the cervical esophagus (about 4 cm in length) underwent esophagoplasty by a new technique following failed attempts at esophagoscopy and dilatation. This procedure results in a neoesophagus in the region with a lumen approximately twice its previous diameter, which will further increase in size gradually after a number of dilatations. The postoperative course was uneventful. She had almost no dysphagia following four successive dilatations 1, 2, 3 and 5 months after surgery. Barium swallow revealed an acceptable lumen. She was asymptomatic 2 years after surgery. PMID- 14566418 TI - The effect of hyperbaric oxygen treatment on the inflammatory changes caused by intraperitoneal meconium. AB - Exposure of the peritoneal cavity to meconium causes a marked inflammatory response. The effect of intraperitoneal meconium on intestinal morphology and plasma nitrite and nitrate (NO2(-) + NO3(-)) levels and how this inflammatory process is influenced by hyperbaric oxygen (HBO) treatment were investigated in this study. The purpose was to determine whether HBO treatment could be considered a useful adjunct in the resuscitative treatment of severely ill patients admitted with meconium peritonitis (MP). Rats were divided into three groups. Human meconium (MP group, n=10) and sterile saline (control group, n=10) were injected intraperitoneally for 3 days. The procedure for meconium injection was combined with HBO treatment for the HBO group (n=10). HBO was administered for 7 days. In all groups, peritoneal swap cultures, plasma NO2(-) + NO3(-) levels, intestinal diameters, and macroscopic and microscopic changes in the intestine were determined on the 8th day. Bacterial growth was not detected in the peritoneal swap cultures. There was a significant difference in NO2(-) + NO3( ) levels between the MP and HBO groups ( P<0.05), between the MP and control groups ( P<0.01), and between the HBO and control groups ( P<0.05). Thin fibrinous adhesions in both the MP and HBO groups, and thickened and dilated intestinal loops in the MP group were observed macroscopically. The intestinal diameter in the MP group was significantly greater than in the HBO and control groups. The only microscopic difference was seen in the serosal layer. Compared with the animals in the control and HBO groups, the intestine of the rats in the MP group showed prominent serosal thickening, edema, capillary proliferation and cellular infiltration. The ameliorated inflammatory changes and decreased dilatation of the intestine accompanied by a significant decrease in NO2(-) + NO3(-) levels suggest that as an adjunctive treatment, HBO may have a beneficial effect in the resuscitative treatment of meconium peritonitis. PMID- 14566419 TI - Glioma after cerebral hydatid disease. AB - BACKGROUND: The authors present the case of a 31-year-old man with a malignant glioma. He had been treated for cerebral hydatid as a child, and 22 years later he developed a glioma at the site of his previous disease. DISCUSSION: Could chronic inflammatory change following intracranial hydatid disease have induced neoplastic transformation of glial cells? PMID- 14566420 TI - The scientific establishment of a new therapeutic intervention for developmental conditions: practical and ethical principles. AB - BACKGROUND: Although the randomized controlled trial (RCT) is a major methodological breakthrough extending the limits of objectivity in clinical medical science, clinical trials of surgery have seldom included placebo surgery as a control, for ethical reasons. Especially in clinical studies intended eventually to establish a new intervention for developmental conditions, it has been recognized that there is huge examiner bias. In addition, the many miraculous cases that have been reported in nonsurgical open trials for developmental conditions and have eventually been evaluated as nonspecific positive outcomes in RCTs suggest that empirically promising interventions must be subjected to scientific scrutiny as soon as possible in the field of developmental conditions. APPLICATION TO PEDIATRIC NEUROSURGERY: Therefore, in childhood neurosurgery, clinical studies to establish a new therapeutic measure for developmental conditions should be designed as rigorously as possible using optimized scientific methods. The worldwide ethical guideline, the Declaration of Helsinki issued by the World Medical Association, can provide principles for the establishment of a new intervention in the treatment of a patient when proven therapeutic methods do not exist or methods used thus far have been ineffective. Physicians' discretion to use unproven or new therapeutic measures for such patients is approved in the presence of efforts of an ethical and scientific approach. Even if the measure is a very promising intervention, the research aspects must completely be demonstrated for informed consent and review by the ethical committee and the trial must be regarded as a clinical research. Especially when an RCT is not possible for ethical reasons, appropriate epidemiological data or animal experiments should suggest that the new measure is effective before a clinical trial. In a clinical setting, where neither epidemiological studies nor animal experimentation can be introduced, if necessary the researcher should collaborate with experts to obtain multidisciplinary justification for clinical testing. PMID- 14566421 TI - Effects of fatigue on the catchlike property in a turtle hindlimb muscle. AB - The purpose of this report was to test for the possibility that a catchlike, force-enhancing property, attributable to a particular stimulation pattern, could be evoked in non-mammalian turtle muscle, just as it has been shown in mammalian muscle. We tested for the presence of this property in dynamic lengthening and shortening contractions, as well as in the more commonly studied isometric contractions. A second aim was to note the effects of fatigue on the catchlike property, if the latter was present. The force response of the external gastrocnemius muscle in the adult turtle, Pseudemys ( Trachemys) scripta elegans, was compared for a control, constant-frequency 10 Hz, 1-s duration stimulation pattern using 0.1-ms pulses vs. the same pattern, but with two additional pulses within the first 100-ms interspike interval of the control stimulus train. This latter train produced a pronounced and prolonged enhancement of muscle force, which was attributed to a catchlike effect. It was greatly increased when the muscle was in a fatigued state. The extent of this force enhancement was significantly different for the three contraction types, and generally in the order: isometric>lengthening>shortening contraction. These differences were greater in fatigued vs. fresh muscle. Comparative aspects and potential mechanisms underlying the catchlike effect are discussed. PMID- 14566422 TI - Wind spectra and the response of the cercal system in the cockroach. AB - Experiments on the cercal wind-sensing system of the American cockroach, Periplaneta americana, showed that the firing rate of the interneurons coding wind information depends on the bandwidth of random noise wind stimuli. The firing rate was shown to increase with decreases in the stimulus bandwidth, and be independent of changes in the total power of the stimulus with constant spectral composition. A detailed analysis of ethologically relevant stimulus parameters is presented. A phenomenological model of these relationships and their relevance to wind-mediated cockroach behavior is proposed. PMID- 14566423 TI - Androgens and male fertility. AB - Androgens play a crucial role in the development of male reproductive organs such as the epididymis, vas deferens, seminal vesicle, prostate and the penis. Furthermore, androgens are needed for puberty, male fertility and male sexual function. High levels of intratesticular testosterone, secreted by the leydig cells, are necessary for spermatogenesis. Intratesticular testosterone is mainly bound to androgen binding protein and secreted into the seminiferous tubules. Inside the sertoli cells, testosterone is selectively bound to the androgen receptor and activation of the receptor will result in initiation and maintenance of the spermatogenic process and inhibition of germ cell apoptosis. The androgen receptor is found in all male reproductive organs and can be stimulated by either testosterone or its more potential metabolite dihydrotestosterone. Severe defects of the androgen receptor may result in abnormal male sexual development. More subtle modulations can be a potential cause of male infertility. Treatment of an infertile man with testosterone does improve spermatogenesis, since exogenous administrated testosterone and its metabolite estrogen will suppress both GnRH production by the hypothalamus and Luteinising hormone production by the pituitary gland and subsequently suppress testicular testosterone production. Also, high levels of testosterone are needed inside the testis and this can never be accomplished by oral or parenteral administration of androgens. Suppression of testosterone production by the leydig cells will result in a deficient spermatogenesis, as can be seen in men taking anabolic-androgenic steroids. Suppression of spermatogenesis by testosterone administration is also the basis for the development of a male contraceptive. During cytotoxic treatment or irradiation suppression of intratesticular testosterone production cells may prevent irreversible damage to the spermotogonial stem cells. PMID- 14566424 TI - Calcified metastatic mediastinal lymph nodes from mucinous breast adenocarcinoma. PMID- 14566425 TI - Improvement of parenchymal and vascular enhancement using saline flush and power injection for multiple-detector-row abdominal CT. AB - The aim of this study was to determine if a saline solution flush following low dose contrast material bolus improves parenchymal and vascular enhancement during abdominal multiple detector-row computed tomography (MDCT). Forty-one patients (24 men and 17 women; mean age 49 years, age range 27-86 years) underwent abdominal MDCT (collimation 4x5 mm, 15-mm table increment, reconstruction interval 5 mm, gantry rotation period 0.8 s) with a single- as well as with a double syringe power injector. Indication for examination were benign and malignant tumors and inflammatory diseases. Patients received 100 ml nonionic contrast material (300 mgI/ml) alone or pushed with 20 ml saline solution. Mean enhancement values for both protocols were measured in the liver, the spleen, the pancreas, the renal cortex, the portal vein, the inferior vena cava and the abdominal aorta. Double syringe power-injector protocol led to significantly higher parenchymal and vascular enhancement than single syringe power-injector protocol (p<0.05). The improvement in mean enhancement of the liver was 9 +/- 9 HU, of the spleen 8 +/- 10 HU, of the pancreas 7 +/- 9 HU, and of the renal cortex 8 +/- 20 HU. The improvement in mean enhancement of the portal vein was 10 +/- 17 HU of the inferior vena cava 8 +/- 13 HU and of the abdominal aorta 10 +/- 17 HU. The use of a double syringe power injector with saline flush following contrast material bolus significantly improves parenchymal and vascular enhancement during contrast-enhanced abdominal MDCT with low iodine doses. PMID- 14566426 TI - Drug smuggling by body packing: what radiologists should know about it. AB - Body packing is a distinct method for smuggling drugs. What radiologists need to know is discussed in this pictorial review. Radiologists are confronted with diagnostic imaging of body packers because of two main reasons: complications of body packing and identifying drug packets within the gastrointestinal tract. The standard examination used is plain X-ray of the abdomen in an upright and a supine position. Computed tomography is occasionally used but nevertheless described as a very accurate diagnostic tool. Ultrasound and MR imaging do not play an important role in that field. Depending on the purity of the drug, three different forms of attenuation have been described: hashish is denser than stool; cocaine appears similar to stool; and heroin has a gaseous transparence. The packets are of a round to oval form, usually of a particular uniformity and rarely confused with scybala if arranged like a pearl chain; therefore, plain X ray is the method of choice to detect drug-filled packets within the gastrointestinal tract of body packers. PMID- 14566427 TI - T2-weighted MR imaging of prostate cancer: multishot echo-planar imaging vs fast spin-echo imaging. AB - The aim of the present study was to assess the performance of pre-biopsy T2 weighted MR imaging using multishot echo-planar imaging (EPI) sequence for visualization of prostate cancer and to compare image quality with that of fast spin-echo (FSE) sequence. Thirty-nine patients with suspected prostate cancer and one healthy male volunteer were examined on a 1.5-T MR scanner equipped with a pelvic phased-array coil. Axial MR images were obtained using multishot EPI sequence with a multishot number of 16 and FSE sequence without fat suppression. Paired EPI and FSE images were independently evaluated by three radiologists. Furthermore, signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were compared between EPI and FSE images of 12 pathologically proven lesions of prostate cancer. Delineation of the periprostatic venous plexus, prostate zonal anatomy, and seminal vesicle on EPI was graded to be superior/inferior to FSE in 15.8/0, 14.6/0, and 21.5/4.3% of cases, respectively. On the other hand, delineation of the neurovascular bundle was superior/inferior to FSE in 2.6/13.2% of cases. The SNR and CNR of prostate cancer on EPI were significantly higher than those on FSE (7.99 +/- 2.51 vs 3.36 +/- 0.58, p<0.0001, and 5.51 +/- 2.02 vs 2.21 +/- 0.79, p<0.0001, respectively). In conclusion, multishot EPI has higher quality of contrast resolution for imaging of prostate cancer compared with FSE and would have the potential usefulness in the detection of prostate cancer, although these results obtained with a phased-array coil cannot be extrapolated to examinations performed with an endorectal coil. PMID- 14566428 TI - Immunization with a plant-produced colorectal cancer antigen. AB - Cancer vaccination has become an important focus of oncology in recent years. Active immunization with tumor-associated antigens such as colorectal cancer antigen GA733-2 is thought to potentially overcome the reoccurrence of metastasis. As recombinant protein production in bioreactors is costly and subject to growing safety concerns, we tested plants as an alternative for the expression of a potential colorectal cancer vaccine. Comparing colorectal cancer antigen GA733-2 produced in tobacco plants with the same antigen produced in insect cell culture, we found a similar humoral immune response to injection of either of the two antigen preparations into mice. Some minor differences were observed in the cellular response that might be due to impurities. Our studies compare for the first time, immunization with the same antigen expressed in either plants or insect cell culture. This will provide important data for use of plants as production systems of therapeutics. PMID- 14566429 TI - Complete lab-scale detoxification of groundwater containing 1,2-dichloroethane. AB - The suspected carcinogenic solvent 1,2-dichloroethane (1,2-DCA) is the most abundant chlorinated C(2) groundwater pollutant on earth. However, an efficient reductive in situ detoxification technology for this compound is not known. Detoxification results of 1,2-DCA with the recently isolated anaerobic bacterium Desulfitobacterium dichloroeliminans strain DCA1 are presented. First, it was verified that strain DCA1 could compete for nutrients in the presence of fast growing Enterococcus faecalis; the latter was observed in the enrichment culture from which strain DCA1 was isolated. Subsequently, lab-scale bioaugmentation of the strain to groundwater containing 40 mg 1,2-DCA/l indicated that the bacterium has strong metabolic activity under prevailing environmental conditions, converting the pollutant into ethene. During exponential growth, the maximum 1,2 DCA dechlorination rate exceeded 350 nmol chloride released per min per mg total bacterial protein. Growth and dechlorination within the community with autochthonous bacteria indicated a high competitive strength of strain DCA1. Interestingly this dechlorination process does not produce any toxic byproducts, such as vinyl chloride. Furthermore, complete groundwater detoxification happens within a short time-frame (days) and is robust in terms of bacterial competition, oxygen tolerance, high ionic strength, and pH range. PMID- 14566430 TI - Bioleaching review part B: progress in bioleaching: applications of microbial processes by the minerals industries. AB - This review describes the historical development and current state of metals leaching and sulfide mineral biooxidation by the minerals industries. During the past 20 years commercial processes employing microorganisms for mineral recovery have progressed from rather uncontrolled copper dump leaching to mineral oxidation and leaching in designed bioheaps for oxidation of refractory gold ores and for copper recovery. Also during this period of time, stirred tank bioleaching has been commercialized for cobalt recovery and for biooxidation of refractory gold ores. Chalcopyrite bioleaching in stirred tanks is on the verge of commercialization. Commercial applications of biohydrometallurgy have advanced due to favorable process economics and, in some cases, reduced environmental problems compared to conventional metal recovery processes such as smelting. Process development has included recognition of the importance of aeration of bioheaps, and improvements in stirred tank reactor design and operation. Concurrently, knowledge of the key microorganisms involved in these processes has advanced, aided by advances in molecular biology to characterize microbial populations. PMID- 14566431 TI - Environmental and cultural stimulants in the production of carotenoids from microorganisms. AB - Commercial production of carotenoids from microorganisms competes mainly with synthetic manufacture by chemical procedures. Efficient stimulation of carotenoid biosynthesis is expected to promote accumulation of carotenoid by microbes. This review describes the variety of environmental and cultural stimulants studied during the last few decades which enhance volumetric production and cellular accumulation of commercially important carotenoids from microalgae, fungi and bacteria. Stimulation of carotenoid production by white-light illumination and temperature fluctuation is discussed along with supplementation of metal ions, salts, organic solvents, preformed precursors and several other chemicals in the culture broth. Reports on the improvements in yield are reviewed and assessed from a biotechnology point of view. PMID- 14566432 TI - Bioleaching review part A: progress in bioleaching: fundamentals and mechanisms of bacterial metal sulfide oxidation. AB - Bioleaching of metal sulfides is caused by astonishingly diverse groups of bacteria. Today, at least 11 putative prokaryotic divisions can be related to this phenomenon. In contrast, the dissolution (bio)chemistry of metal sulfides follows only two pathways, which are determined by the acid-solubility of the sulfides: the thiosulfate and the polysulfide pathway. The bacterial cell can effect this sulfide dissolution by "contact" and "non-contact" mechanisms. The non-contact mechanism assumes that the bacteria oxidize only dissolved iron(II) ions to iron(III) ions. The latter can then attack metal sulfides and be reduced to iron(II) ions. The contact mechanism requires attachment of bacteria to the sulfide surface. The primary mechanism for attachment to pyrite is electrostatic in nature. In the case of Acidithiobacillus ferrooxidans, bacterial exopolymers contain iron(III) ions, each complexed by two uronic acid residues. The resulting positive charge allows attachment to the negatively charged pyrite. Thus, the first function of complexed iron(III) ions in the contact mechanism is mediation of cell attachment, while their second function is oxidative dissolution of the metal sulfide, similar to the role of free iron(III) ions in the non-contact mechanism. In both cases, the electrons extracted from the metal sulfide reduce molecular oxygen via a complex redox chain located below the outer membrane, the periplasmic space, and the cytoplasmic membrane of leaching bacteria. The dominance of either At. ferrooxidans or Leptospirillum ferrooxidans in mesophilic leaching habitats is highly likely to result from differences in their biochemical iron(II) oxidation pathways, especially the involvement of rusticyanin. PMID- 14566433 TI - SulA-independent filamentation of Escherichia coli during growth after release from high hydrostatic pressure treatment. AB - To improve the efficiency of sterilization by high hydrostatic pressure treatment (HPT), it is desirable to know the biochemical process of bacteria most sensitive to the treatment. We investigated growth properties after release from HPT of exponentially growing Escherichia coli K-12 cells. We observed growth retardation after treatment (30 min at 37 degrees C) above 75 MPa. Long filamentous cells of about eight times normal cell length were observed at 90 min growth after treatment at 75 MPa. In the subsequent period the filamentous cells divided into normal-sized cells. recA and sulA mutant strains also formed filamentous cells, indicating that filamentation was SulA-independent. Nucleoids segregated normally in the filamentous cells. Only one FtsZ ring (or none) was detected at possible division sites in the elongated cells. Western blotting analysis demonstrated that the amount of FtsZ protein was not affected by the treatment. GTP-dependent in vitro polymerization of either FtsZ protein in E. coli crude extract or purified FtsZ protein, however, was sensitive to HPT. These facts suggest that HPT at 75 MPa denatures a fraction of FtsZ molecules, and that these denatured molecules interfere with the polymerization of functional FtsZ, resulting in the significantly reduced number of FtsZ rings. PMID- 14566434 TI - Evolutionary stability of MHC class II haplotypes in diverse rhesus macaque populations. AB - A thoroughly characterized breeding colony of 172 pedigreed rhesus macaques was used to analyze exon 2 of the polymorphic Mamu- DPB1, -DQA1, -DQB1, and - DRB loci. Most of the monkeys or their ancestors originated in India, though the panel also included animals from Burma and China, as well as some of unknown origin and mixed breeds. In these animals, mtDNA appears to correlate with the aforementioned geographic origin, and a large number of Mamu class II alleles were observed. The different Mamu- DPB1 alleles were largely shared between monkeys of different origin, whereas in humans particular alleles appear to be unique for ethnic populations. In contrast to Mamu-DPB1, the highly polymorphic - DQA1/DQB1 alleles form tightly linked pairs that appear to be about two-thirds population specific. For most of the DQA1/DQB1 pairs, Mamu- DRB region configurations present on the same chromosome have been ascertained, resulting in 41 different -DQ/DRB haplotypes. These distinct DQ/DRB haplotypes seem to be specific for monkeys of a determined origin. Thus, in evolutionary terms, the Mamu-DP, -DQ, and -DR regions show increasing instability with regard to allelic polymorphism, such as for -DP/DQ, or gene content and allelic polymorphism, such as for -DR, resulting in population-specific class II haplotypes. Furthermore, novel haplotypes are generated by recombination-like events. The results imply that mtDNA analysis in combination with Mhc typing is a helpful tool for selecting animals for biomedical experiments. PMID- 14566435 TI - The IDDM-associated solitary retroviral promoters DQ-LTR3 and DQ-LTR13 have a distinct impact on the expression of selected DQB1 genes in different cell lines in vitro. AB - The HLA complex on chromosome 6, especially of the HLA class II genes, plays an important role in the pathogenesis of insulin-dependent diabetes mellitus (IDDM). Three solitary long terminal repeats (LTRs) have been described in the vicinity of HLA DQ, two of them modifying the genetic susceptibility to type 1 diabetes on high-risk HLA DQ haplotypes. Therefore, we have investigated the putative regulatory properties of these retroviral promoters in different cell lines, including a number of B- and T-lymphoblastoid cell lines bearing different DQ haplotypes, employing a transient reporter gene assay. The transcriptional activity of appropriate constructs harboring an LTR linked to the luciferase reporter gene revealed different expression patterns which varied considerably between the investigated cell lines. DQ-LTR3 showed clear activities, whereby the levels of luciferase gene expression were also increased approximately 200-fold in the teratocarcinoma cell line GH. For the different B- and T-cell lines, no significant activity was detected for any of the investigated LTRs. Furthermore, we have analyzed the effect of DQ-LTR13 on distinct DQB1 promoters and could show an increased activity of the DQB1*0302 promoter under the influence of DQ-LTR13. It varied from 1.5- to 6-fold in different cell lines depending on the transcriptional orientation and position of the LTR and was independent of DQ LTR3. Moreover, the impact of the LTR was different for the DQB1*0201 promoter demonstrating a decreasing effect. These data indicate that a DQ-LTR13-mediated impact on the DQB1 promoter activity exists which differs clearly between selected promoters. PMID- 14566436 TI - Physical and genetic mapping of the rainbow trout major histocompatibility regions: evidence for duplication of the class I region. AB - One of the most unexpected discoveries in MHC genetics came from studies dealing with the teleost MHC. Initially discovered in zebrafish, the MHC class I and II regions of all bony fish are not linked. Previous segregation analysis in trout suggested that the class I and II regions reside on completely different chromosomes. To learn more about MHC genomics in trout, we have isolated BAC clones harboring class Ia and Ib loci, a single BAC clone containing an MH class II gene ( DAB), as well as BAC clones containing the ABCB2 gene. Upon PCR and sequence confirmation, BAC clones were labeled and used as probes for in situ hybridization on rainbow trout metaphase chromosomes for determination of the physical locations of the trout MH regions. Finally, SNPs, RFLPs, and microsatellites found within the BAC clones allowed for these regions to be assigned to specific linkage groups on the OSU x Hotcreek (HC) and OSU x Arlee (ARL) genetic linkage maps. Our data demonstrate that the trout MH regions are located on at least four different chromosomes and the corresponding linkage groups, while also providing direct evidence for the partial duplication of the MH class I region in trout. PMID- 14566437 TI - Radiological appearance of diaphragmatic mesothelial cysts. AB - BACKGROUND: Diaphragmatic mesothelial cysts are congenital lesions that derive from coelomic remnants. OBJECTIVE: To describe their radiological appearance on US, CT and MRI. MATERIALS AND METHODS: Four children (aged 7-14 years) who underwent surgery and for whom there was pathological confirmation of the diagnosis. Five more children (0-12 months old) who were managed conservatively, but for whom there was a high degree of suspicion for this diagnosis. RESULTS: All lesions presented as a cystic mass between the thoracic wall and the liver, in the right posterolateral costophrenic angle. Some lesions showed subtle evidence of their extrahepatic location. CONCLUSIONS: The radiological findings identified are highly suggestive of diaphragmatic mesothelial cyst. PMID- 14566438 TI - Metatropic dysplasia lethal variants. AB - BACKGROUND: The metatropic dysplasia group includes fibrochondrogenesis, Schneckenbecken dysplasia and metatropic dysplasia (various forms). The overlapping features of this group with other dysplasias may cause diagnostic confusion, particularly in perinatal lethal cases. OBJECTIVE: To attempt to classify the radiological findings of the presented eight sporadic cases based on a broad review of the perinatally lethal metatropic group of conditions and to discuss some overlapping features in the light of current knowledge. RESULTS: The first four cases are of recognised conditions, namely lethal metatropic dysplasia (Type 2) or hyperchondrogenesis, lethal hyperplastic metatropic dysplasia (Type 1) and fibrochondrogenesis. The remaining four cases cannot be categorised accurately and are different from each other but with some features of the metatropic group of dysplasias. CONCLUSIONS: The dysplasias within the metatropic dysplasia group are phenotypically distinct from many forms of chondrodysplasia but the pathogenesis still remains poorly understood from the morphological and molecular perspectives. Chondro-osseous morphology might be helpful in all lethal cases especially in our last four cases. PMID- 14566439 TI - Recessive omodysplasia: five new cases and review of the literature. AB - BACKGROUND: Autosomal recessive omodysplasia (MIM 258315) is a rare skeletal dysplasia characterized by severe congenital micromelia with shortening and distal tapering of the humeri and femora to give a club-like appearance. Fewer than 20 cases have been reported in the literature so far. OBJECTIVE: The purpose of this study was to more clearly describe the clinical and radiographic phenotypes and their changes with age. MATERIALS AND METHODS: Five new patients, including two sibs, with autosomal recessive omodysplasia are presented. RESULTS: Clinical features are rhizomelic dwarfism with limited extension of elbows and knees and a distinct face with a short nose, depressed nasal bridge, long philtrum, midline haemangiomas in infants and cryptorchidism in males. Radiological findings are distal hypoplasia of the short humerus and femur with characteristic radial dislocation and radioulnar diastasis. CONCLUSIONS: Based on a review of these and 16 previously reported patients, the regressive nature of the humerofemoral changes and the obvious male predominance are stressed. Phenotypic similarities with the atelosteogenesis group of disorders and with diastrophic dysplasia suggest common pathogenetic mechanisms. PMID- 14566440 TI - Population pharmacokinetics of gentamicin in South African newborns. AB - OBJECTIVE: Gentamicin population pharmacokinetics in newborns were studied with special reference to possible gender effects. METHODS: Steady-state serum levels ( n=139) were obtained from 79 neonates with a mean birth weight of 2.1 kg, mean gestational age of 35.1 weeks and mean age at the time of sampling of 4.2 days. The data were analysed using the non-linear mixed effects model (NONMEM). A one compartment model was used to fit the data. RESULTS: The final models for clearance (CL) and volume of distribution (V) were: CL(l/h)=0.001xWGTxGAxP and V(l)=0.472xWGT, where WGT=birth weight (kg), GA=gestational age (weeks) and P=1.2 for girls and 1.0 for boys. The values of inter-individual variability in CL and V were 34% and 35%, respectively. Intra-individual variability was 5% (proportional) and 7.2% (additive). Mean (95% confidence interval) values of CL and half-life were 0.042 l h(-1) kg(-1) (0.041, 0.043 l h(-1) kg(-1)) and 8.0 h (7.7, 8.3 h), while V was 0.472 (0.428, 0.516) l/kg for all patients. CONCLUSION: Mean population pharmacokinetic values were similar to those obtained with NONMEM for gentamicin in other neonates of similar age. Gender was found to be a determinant of CL, with girls clearing faster than boys. PMID- 14566441 TI - The delusion of reducing sample size. PMID- 14566442 TI - Effect of the novel anxiolytic drug deramciclane on the pharmacokinetics and pharmacodynamics of the CYP3A4 probe drug buspirone. AB - RATIONALE: Preliminary in vitro findings indicated that the novel anxiolytic drug, deramciclane is a substrate for the cytochrome P(450) (CYP) 3A4 isoenzyme. Moreover, its co-administration with buspirone, another anxiolytic drug, is likely in clinical practice. OBJECTIVES: The primary objective of the present study was to evaluate the in vivo effects of deramciclane on CYP3A4 activity as measured by buspirone pharmacokinetics. The secondary objective was to study the possible pharmacodynamic interaction between these two anxiolytic drugs. METHODS: Sixteen healthy subjects received 60 mg deramciclane or matched placebo for 8 days in this randomized, double-blind, cross-over study. On day 8 of both phases, the subjects received a 20-mg single dose of buspirone. Buspirone and its active metabolite, 1-pyrimidylpiperazine (1-PP), concentrations were measured for 24 h. Pharmacodynamic testing and measurement of plasma prolactin concentrations were carried out on day 7 and day 8 to assess the pharmacodynamic consequences of deramciclane and buspirone co-administration. RESULTS: Repeated administration of deramciclane had no effect on CYP3A4 activity as measured by buspirone pharmacokinetics. However, deramciclane administration caused an inhibition of the further, not CYP3A4-dependent, metabolism of 1-PP as evidenced by 84% increase in the AUC ( P<0.001) and 20% increase in the elimination half-life ( P=0.0012) of 1-PP. Deramciclane did not potentiate the buspirone-induced increase in prolactin secretion. No significant differences were found in the psychomotoric testing or the subjective maximum sedation between the deramciclane phase and the placebo phase, either before or after buspirone administration. Of 16 subjects, 5 experienced dizziness during both study phases. CONCLUSION: Deramciclane does not inhibit CYP3A4 activity as measured by buspirone pharmacokinetics, and there were no indications of relevant pharmacodynamic interaction after multiple doses of deramciclane and a single dose of buspirone. PMID- 14566443 TI - Error reduction: academic detailing as a method to reduce incorrect prescriptions. AB - OBJECTIVES: Clinical decision making can be influenced by academic detailing, and it was hypothesised that this technique may be used to reduce simple errors when prescribing drugs of addiction. These prescriptions require specific details to be included, otherwise the prescription has to be returned to the prescriber and re-written, wasting the time of the dispenser and prescriber alike. METHODS: The legal requirements for the prescription of addictive drugs were used to define prescription errors. Prescription error rates at six hospitals were assessed, including a control and an intervention hospital where academic detailing was carried out. Prescription error rates were documented before and after an academic detailing visit to junior doctors, including the provision of a bookmark containing the requirements for these drugs. These errors were expressed as a percentage of the total written for a 4-week period. RESULTS: At the intervention hospital, there was a significant decrease in error rate (from 41% to 24%, P<0.0001) with an improvement in all the requirements stated on the bookmark. At this hospital, the confidence of the junior doctors on a self-rating 5-point scale in writing prescriptions for these drugs increased from a mean of 3.25 (95% CI 2.92-3.58) to 4.14 (95%CI 3.90-4.38) after the intervention (P=0.03). The baseline error rates at the other hospitals ranged from 25% to 46%, but the control hospital did not show a significant change in error rate over the same study period ( P=0.66). A post-hoc review suggested that liquid preparations were more difficult to prescribe correctly, which in part accounted for the differences in error rate between hospitals. CONCLUSIONS: Academic detailing appears to be a useful method of reducing erroneous hospital prescriptions; and, to our knowledge, this is a novel finding. PMID- 14566444 TI - The neural control of single degree-of-freedom elbow movements. Effect of starting joint position. AB - It is known that muscle activation patterns are changed when the force requirements of a task are increased (e.g., moving a heavier inertial load) or when the available muscle force is reduced (e.g., by inducing muscle fatigue). It is not known whether this is true when the torque-producing capability of a muscle is altered. Eight neurologically healthy subjects performed flexion and extension maximum voluntary isometric contractions (MVC) at five different joint positions (10 degrees, 40 degrees, 70 degrees, 100 degrees, and 130 degrees, where 0 degrees is full elbow extension). Flexion MVC increased by 138% and extension MVC increased by 74% as the elbow joint position changed from the most extended to the most flexed position tested. The same subjects then made rapid, 30 degrees elbow flexion movements from each of four starting elbow positions (10 degrees, 40 degrees, 70 degrees, and 100 degrees). Muscle activation patterns for movements made from the more extended positions showed an increased first agonist burst duration and increased latency of the antagonist burst. There was no change in the initial rate of rise of the agonist burst across starting joint positions. Movements made from the most extended starting position were significantly slower and had longer acceleration and deceleration times than did movements made from the more flexed starting positions. The changes in muscle activation patterns were consistent with those seen when the force requirements of a task are increased or the available muscle force is reduced. We hypothesize that a fall in the ratio of available to required muscle forces causes the nervous system to change muscle activation patterns, to increase the ratio. Our results are consistent with this hypothesis. PMID- 14566446 TI - Task-dependent motor learning. AB - We examined whether task-dependent modulation was evident in a motor learning paradigm. Subjects performed reaching movements before, during, and after exposure to a novel force perturbation while adopting either a spatial goal, "continue towards the target", or an effort goal, "keep your effort profile the same". Before the perturbation, the hand trajectories were moderately straight and accurate regardless of the task. However, during and immediately after the perturbation, the reaches exhibited unambiguous task-dependent differences in both the initial and terminal periods of the reach. With the spatial goal, subjects showed terminal compensations to the force-induced displacements indicative of feedback control. In addition, feedforward control was evident in the smaller path deviations with continued exposure and the initial path aftereffects when the perturbation was removed. In contrast, when adopting an effort goal, subjects showed large and chronically deviated endpoints from the perturbation indicating an absence of feedback compensation. They also showed no feedforward adaptation during repeated exposure or visible aftereffects when the perturbation was removed. Therefore, both feedforward and feedback control mechanisms show task-dependent modulation in a motor learning paradigm. PMID- 14566445 TI - Neuromaturation of human locomotion revealed by non-dimensional scaling. AB - When two fundamental gait parameters--step length and step frequency--are scaled non-dimensionally, thereby accounting for increases in a child's physical size, ontogenetic changes in the locomotor control strategy are revealed. We believe dimensionless velocity beta, the product of dimensionless step length and frequency, serves as a measure of neural development. It increases from the age of 18 months and reaches a plateau between 50 and 90 months, attaining the adult value of 0.45. Based on a study of 200 children, our findings lend support to a theory that posits a neuromaturation growth curve with the form: beta (t)=0.45 (1 e(-0.05t)) where t is the age in months and 0.05 is the growth coefficient. PMID- 14566447 TI - Reward potentiation or behavioral activation? A comment on Donny et al. PMID- 14566449 TI - Interleukin-1beta mediates endotoxin- and tumor necrosis factor alpha-induced RGS16 protein expression in cultured cardiac myocytes. AB - Endotoxin (LPS)-induced cardiac failure is associated with an up-regulation of RGS16 protein expression and repression of phospholipase C activity in vivo. Since the release of pro-inflammatory cytokines plays an important role in mediating LPS-induced myocardial dysfunction, we examined the effect of recombinant cytokines on the expression of RGS16 protein in neonatal cardiac myocytes. Myocytes in culture were treated with 50 ng/ml recombinant tumor necrosis factor alpha (TNFalpha), 2 ng/ml interleukin 1beta (IL-1beta), interleukin 6 (IL-6), interferon gamma (IFNgamma) or diluent (NaCl) for 24 h. Before stimulation with LPS (4 micro g/ml for 24 h) cells were treated with 200 ng/ml interleukin 1-receptor antagonist (IL-1ra), 500 ng/ml soluble TNF receptor (sTNFr), or NaCl for 1 h. Isolated membrane proteins were used for Western blot analysis. Cell-associated and secreted IL-1beta and TNFalpha protein content were determined in myocyte protein homogenates and cell culture supernatants by ELISA immunoblotting 3, 6, 24, 48 and 72 h after treatment with LPS. IL-1beta (1.75 fold) and TNFalpha (1.62-fold) but not IL-6 and IFNgamma induced RGS16 protein expression. LPS stimulated intracellular IL-1beta expression within 6 h (847.1+/ 172.9 pg/3x10(6) cells) followed by an increase in extracellular secretion up to 70.8+/-8.1 pg/3x10(6) cells after 48 h. In contrast, intracellular protein concentrations of TNFalpha were almost not detectable (0.03+/-0.01 pg/3x10(6) cells), but extracellular secretion was induced by LPS with a maximum at 6 h (653.9+/-36.3 pg/3x10(6) cells). The LPS-induced increase in RGS16 (1.6-fold) was blunted by IL-1ra but not by TNFalpha scavenging. Interestingly, both, the IL 1beta- and TNFalpha-effect could be blocked by IL-1ra, indicating that also the TNFalpha-induced RGS16 expression is mediated by IL-1. We therefore conclude that LPS induces RGS16 protein expression by activation of the cytokine IL-1beta in cardiac myocytes. Our data substantiate the role of IL-1beta as an important mediator in LPS-induced cardiac failure. PMID- 14566450 TI - Quantitative measurement of depolarization-induced anandamide release in human and rat neocortex. AB - It has been suggested that the endocannabinoid anandamide is released from central neurons upon depolarization of the cell membrane. In order to determine whether anandamide levels were increased after K(+) depolarization of fresh human and rat brain slices, we developed a rapid and sensitive method for the isolation and quantitation of anandamide. This included solvent extraction, solid phase separation, and reversed phase high performance liquid chromatography (HPLC) with fluorometric detection. Compared with basal levels, K(+) stimulation enhanced the neocortical anandamide concentration in both species (70.5 vs. 21.1 pmol/g tissue in humans, 14.3 vs. 3.2 pmol/g tissue in rats). Basal anandamide levels in the rat hippocampus (11.1 pmol/g) were significantly higher than in the neocortex. Anandamide was also detected in the human amygdala (67.8 pmol/g). In conclusion, our data provide evidence for the depolarization-induced synthesis of anandamide, supporting the hypothesis of a neuromodulatory action of this endocannabinoid. Furthermore, the presence of anandamide in the limbic system suggests participation in cognition, behavior or reward. PMID- 14566451 TI - The alpha 2-adrenoceptor antagonist atipamezole potentiates anti-Parkinsonian effects and can reduce the adverse cardiovascular effects of dopaminergic drugs in rats. AB - The present experiments investigated the effects of the specific alpha(2) adrenoceptor antagonist atipamezole, alone and in combination with a dopamine agonist, on motor function in rats with a unilateral 6-hydroxydopamine lesion of the nigro-striatal pathway and on exploratory behaviour and cardiovascular function in rats equipped with telemetry transmitters. Dexmedetomidine, an alpha(2)-adrenoceptor agonist and the alpha(2)-adrenoceptor antagonists idazoxan and yohimbine were used as reference compounds. In the unilaterally lesioned animals, direct dopamine agonists, such as apomorphine, induce contralateral turning behaviour. Indirect agonists, such as amphetamine, induce ipsilateral circling in the animals. Atipamezole (0.3 mg/kg s.c) potentiated and dexmedetomidine (10 micro g/kg s.c.) decreased contralateral circling evoked by apomorphine (50 micro g/kg s.c.) and by l-3,4-dihydroxyphenylalanine (L-DOPA, 5 mg/kg i.p.). Atipamezole also prolonged the duration of action of L-DOPA. Atipamezole dose-dependently induced ipsilateral turning behaviour and potentiated turning induced by amphetamine (1 mg/kg i.p.). The alpha(1) adrenoceptor antagonist prazosin (0.1 mg/kg i.p.) partially antagonised the effect of amphetamine and had a strong inhibitory effect on the atipamezole induced potentiation of the amphetamine response. Prazosin did not have any major effect on either the apomorphine response itself or on the potentiation of the apomorphine response by atipamezole. This suggests that atipamezole can modulate motor function both indirectly, by stimulating the release of noradrenaline and directly, by blocking postsynaptic alpha(2)-adrenoceptors in neurones other than noradrenergic nerves. The alpha(2)-adrenoceptor antagonists, when tested at comparably effective central alpha(2)-adrenoceptor antagonising doses in a rat mydriasis model: atipamezole 0.3 mg/kg s.c., idazoxan 1 mg/kg s.c. and yohimbine 3 mg/kg s.c., all induced ipsilateral turning behaviour and potentiated apomorphine-induced contralateral circling. The effects of the alpha(2) adrenoceptor antagonists were in general similar in these experiments. In habituated non-lesioned rats equipped with telemetry transmitters, apomorphine (50 micro g/kg s.c.) decreased blood pressure in the home cage and in an open field test. It also decreased spontaneous motor activity in the open field. Neither atipamezole (0.3 mg/kg s.c.) nor idazoxan (1 mg/kg s.c.) had any effect on blood pressure when given alone, but reversed the apomorphine-induced decrease in blood pressure. Atipamezole also diminished apomorphine-induced sedation in the open-field test. In conclusion, atipamezole improved the efficacy of L-DOPA and apomorphine in an animal model of Parkinson's disease and also reduced adverse dopaminergic effects on vigilance and on cardiovascular function. These results suggest that an investigation of the effects of specific alpha(2) adrenoceptor antagonists in Parkinson's disease patients is warranted. PMID- 14566453 TI - Isolated complete popliteal artery rupture associated with knee dislocation. Case reports. AB - Complete dislocation of the knee is a rare injury, and is frequently associated with injuries to other structures in the popliteal fossa. Prompt recognition of associated popliteal artery disruption and early revascularization is paramount for successful and functional results. The necessity for emergency vascular reconstruction markedly complicates an already difficult orthopaedic problem. Arteriography is recommended in all cases of complete dislocation of the knee. PMID- 14566452 TI - CB1- and CB2-cannabinoid receptor-independent lipolysis induced by WIN 55,212-2 in male rat adipocytes. AB - The expression of genes encoding the cannabinoid CB(1) and CB(2) receptors and fatty acid amide hydrolase (FAAH) and the lipolytic activity of cannabinoid agonists were investigated in rat adipose tissue.RT-PCR studies indicated that the genes encoding CB(1) and CB(2) receptors and FAAH are not expressed in epididymal adipocytes. In functional studies, the non-selective cannabinoid receptor agonist WIN 55,212-2 concentration-dependently (0.01-30 micro M) induced glycerol release above baseline ( E(max) 96.1+/-6.2% of isoprenaline-induced lipolytic response). The selective CB(2) agonist JWH-015 (0.01-30 micro M) had no lipolytic activity while the endocannabinoid 2-arachidonoylglycerol and the stable anandamide derivative, R(+)-methanandamide had, only a weak lipolytic effect at the highest concentrations employed (10 and 30 micro M). The concentration/response relationship for WIN 55,212-2-mediated lipolytic activity, mimicked by the S(-)-enantiomer WIN 55,212-3, was shifted significantly to the right by the CB(1) antagonist AM 251 only at 10 micro M, but was not modified by the beta-adrenoceptor antagonist propranolol (1 micro M). The protein kinase inhibitor H-89, but not the two adenylyl cyclase inhibitors (+/-) N(6)- R phenylisopropyladenosine (R-PIA, 1 micro M, a selective A(1) adenosine receptor agonist) or SQ 22,536 (50 micro M) significantly reduced the glycerol efflux induced by WIN 55,212-2. Our data suggest that the cannabinoid drug WIN 55,212-2 may exert lipolytic activity in male rat adipocytes via an intracellular mechanism, not activated by CB(1) or CB(2) receptor stimulation, significantly reversed by H-89 but not clearly linked to stimulation of adenylyl cyclase. PMID- 14566454 TI - Searching for evidence: don't forget the foundations. PMID- 14566455 TI - Clinical pulmonary infection score for ventilator-associated pneumonia: accuracy and inter-observer variability. AB - OBJECTIVE: Although quantitative microbiological cultures of samples obtained by bronchoscopy are considered the most specific tool for diagnosing ventilator associated pneumonia, this labor-intensive invasive technique is not widely used. The Clinical Pulmonary Infection Score (CPIS), a diagnostic algorithm that relies on easily available clinical, radiographic, and microbiological criteria, could be an attractive alternative for diagnosing ventilator-associated pneumonia. Initially, the CPIS scoring system was validated upon 40 quantitative cultures of bronchoalveolar lavage fluid from 28 patients, and only few other studies have evaluated this scoring system since then. Therefore, little is known about the accuracy of this score. DESIGN: We compared the scores of a slightly adjusted CPIS with results from quantitative cultures of bronchoalveolar lavage fluid in 99 consecutive patients with suspicion of ventilator-associated pneumonia, using growth of > or =10(4) cfu/ml in bronchoalveolar lavage fluid as a cut-off for diagnosing ventilator-associated pneumonia. In addition, the CPIS were calculated for 52 patients by two different intensivists to determine the inter-observer variability. RESULTS: Ventilator-associated pneumonia was diagnosed in 69 (69.6%) patients. When using a CPIS >5 as diagnostic cutoff, the sensitivity of the score was 83% and its specificity was 17%. The area under the Receiver Operating Characteristic curve was 0.55. The level of agreement for prospectively measured Clinical Pulmonary Infection Score (< or =6 and >6) was poor (kappa =0.16). CONCLUSIONS: When compared to quantitative cultures of bronchoalveolar lavage fluid, the CPIS has a low sensitivity and specificity for diagnosing ventilator associated pneumonia with considerable inter-observer variability. PMID- 14566456 TI - HDL-cholesterol level and cortisol response to synacthen in critically ill patients. AB - OBJECTIVE: To explore the relationship between cholesterol levels and the adrenal cortisol response to synacthen in critically ill patients. DESIGN: Prospective observational study. PATIENTS: Critically ill patients with multiple organ dysfunction syndrome (MODS) with possible adrenal dysfunction defined as unexplained hypotension, ongoing inotropic support, unexplained fever, unexplained hyponatraemia or a combination of these symptoms. MEASUREMENTS: HDL cholesterol levels (HDL), total cholesterol levels (TC), and triglycerides (TG) before administration of synacthen. LDL-cholesterol was calculated using the Friedewald formula. Basal cortisol and response to 250 microg synacthen intravenously was measured. A cortisol rise of 0.25 micromol/l in a 30-min or 60 min blood sample after synacthen infusion was defined as a proper adrenal response. RESULTS: Patients with a proper response to synacthen showed higher HDL cholesterol levels than patients without that response ( P=0.02). Severity of disease as measured by APACHE II or SOFA was not a confounder. LDL-cholesterol levels were extremely low in both responders and non-responders and were not associated with the absolute rise in cortisol. In linear and logistic regression analysis HDL-cholesterol was the sole predictor of cortisol response. CONCLUSIONS: Adrenal cortisol response to a "classic" 250-microg synacthen test relates in critically ill patients to HDL-cholesterol levels. LDL and TC levels did not show such a relation. These findings are in concordance with known biochemical pathways of cortisol production. PMID- 14566457 TI - Electrical impedance tomography: a method for monitoring regional lung aeration and tidal volume distribution? AB - OBJECTIVE: To demonstrate the monitoring capacity of modern electrical impedance tomography (EIT) as an indicator of regional lung aeration and tidal volume distribution. DESIGN AND SETTING: Short-term ventilation experiment in an animal research laboratory. PATIENTS AND PARTICIPANTS: One newborn piglet (body weight: 2 kg). INTERVENTIONS: Surfactant depletion by repeated bronchoalveolar lavage, surfactant administration. MEASUREMENTS AND RESULTS: EIT scanning was performed at an acquisition rate of 13 images/s during two ventilatory manoeuvres performed before and after surfactant administration. During the scanning periods of 120 s the piglet was ventilated with a tidal volume of 10 ml/kg at positive end expiratory pressures (PEEP) in the range of 0-30 cmH(2)O, increasing and decreasing in 5 cmH(2)O steps. Local changes in aeration and ventilation with PEEP were visualised by EIT scans showing the regional shifts in end-expiratory lung volume and distribution of tidal volume, respectively. In selected regions of interest EIT clearly identified the changes in local aeration and tidal volume distribution over time and after surfactant treatment as well as the differences between stepwise inflation and deflation. CONCLUSIONS: Our data indicate that modern EIT devices provide an assessment of regional lung aeration and tidal volume and allow evaluation of immediate effects of a change in ventilation or other therapeutic intervention. Future use of EIT in a clinical setting is expected to optimise the selection of appropriate ventilation strategies. PMID- 14566458 TI - Temporal changes in muscle glutathione in ICU patients. AB - OBJECTIVE: This study investigated the changes over time in glutathione and its constituent amino acids in skeletal muscle of ICU patients with multiple organ failure. DESIGN AND SETTING: Prospective and descriptive pilot study in two medium-sized ICUs with ten beds. PATIENTS: Critically ill patients ( n=10) with multiple organ failure and with an expected ICU stay longer than 6 days were included during their initial 3 days after admission to the ICU. MEASUREMENTS AND RESULTS: Muscle biopsy and blood samples were taken on days 0, 3, and 6 after inclusion and total, reduced, and oxidized glutathione and the related amino acids were determined. During the study period both total and reduced glutathione increased and was in the normal range on day 6. The constituent amino acids normalized during the study period as well. CONCLUSIONS: This pilot study demonstrates a recovery of muscle glutathione concentrations in critically ill patients with ongoing multiple organ failure within 1 week. Restoration of muscle glutathione seems to be a biological process of high priority in this group of patients. PMID- 14566459 TI - Validation of the multiple organ dysfunction (MOD) score in critically ill medical and surgical patients. AB - OBJECTIVE: To validate the Multiple Organ Dysfunction (MOD) score externally. DESIGN: Prospective observational cohort study. SETTING: Mixed medical/surgical ICU in a tertiary referral university hospital. PATIENTS AND PARTICIPANTS: Thousand eight hundred and nine patients admitted to ICU for more than 24 h over a 3-year period. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: The MOD score was calculated daily for all patients. The criterion validity of the individual organ scores, the maximal MOD score and the change in MOD score were assessed by examining the relationship between increasing scores and ICU mortality. Increased maximal MOD scores and each of the six individual organ scores, and change in MOD scores were associated with increased mortality. CONCLUSIONS: Maximal and individual organ scores have criterion validity when tested in a different ICU from that in which the scores were derived, indicating that the scoring systems are reproducible. The association of change in MOD score with mortality indicates that the score is responsive. These data, combined with previous data establishing concept and content validity, indicate that the MOD score is a valid measure of multi-organ dysfunction. PMID- 14566460 TI - [Pathogenetic aspectics of nephrotic syndrome]. AB - Nephrotic syndrome is characterized by protein loss in the urine, hypoalbuminemia, hyperlipidemia and edema. Several diseases cause a nephrotic syndrome, as they damage the glomerular podocytes. These specialized epithelial cells, together with endothelial cells of the glomerular capillaries and the basal membrane, form a filter that retains plasma proteins in the circulation. A disturbance of this filter causes proteinuria. The three most common primary glomerular diseases are minimal change, membranous glomerulonephritis, and the primary focal segmental glomerulosclerosis. The familiar forms are rare; however, the identification of the relevant gene defects has greatly advanced our understanding of podocyte function as well as the pathogenesis of nephrotic syndrome. PMID- 14566461 TI - [Treatment of glomerulonephritis]. AB - The treatment of primary glomerulonephritis is a complex matter because of an unclear clinical picture. Glomerulonephritis may emerge as acute nephritis, nephrotic syndrome or minor proteinuria or hematuria. The symptomatic treatment may be derived from the clinical status; immunosuppressive therapy has to be substantiated by renal biopsy in order to offer the best choice to the patient.Rapid-progressive glomerulonephritis must be treated aggressively, as early as possible, to prevent chronic renal failure. Nephrotic syndrome should be treated symptomatically. Immunosuppressants are indicated according to the histological picture and accompanying clinical risk factors for progressive renal disease, which have to be evaluated before treatment. This paper gives the current strategies for treating primary glomerulonephritis. PMID- 14566462 TI - [Minimal Change Glomerulonephritis]. AB - We report two cases of minimal change glomerulonephritis (synonyms: idiopathic nephrotic syndrom, minimal change disease). A 47-year old female patient was admitted to our unit with a relapsing nephrotic syndrome since childhood. Another patient, a 22-year old female, presented with moderately swollen legs that developed over several months and a complaint of frequent upper respiratory tract infections during the last year. In both cases we suspected a minimal change glomerulonephritis which can only be proven by renal biopsy. Therapeutic options comprise steroids, cyclosporin, tacrolimus or even cyclophosphamide, depending on the clinical presentation of the disease in the individual case. PMID- 14566463 TI - [Virus associated glomerulonephritis]. AB - Virus associated glomerulonephritis is considered to be a para- or post infectious autoimmune phenomena. The disease is mediated by immune complexes which usually contain the viral antigen. Virus associated glomerulonephritis due to chronic viral infection with hepatitis B or C virus, or with human immunodeficiency virus (HIV) shows a typical histomorphological picture for each virus. Hepatitis B virus usually leads to a membranous glomerulonephritis, while hepatitis C virus is associated with a membranoproliferative glomerulonephritis due to cryoglobulins, and HIV is associated with a focal segmental sclerosing glomerulonephritis. Knowledge of the relationship between the primary viral infection and secondary glomerulonephritis is important, as a primary immunosuppressive therapy might lead to more severe viral disease. On the other hand, a primary reduction in the viral load due to antiviral therapy with immunostimulants or inhibitors of viral replication could lead to an amelioration of the secondary glomerulonephritis. PMID- 14566464 TI - [Rapidly progressive glomerulonephritis:classification, pathogenesis and clinical management]. AB - Rapidly progressive glomerulonephritides (RPGN) belong to a heterogeneous group of inflammatory kidney diseases which are commonly associated with systemic vasculitic syndromes. Renal histology is characterized by necrotizing lesions within the glomerual tuft and extracapillary proliferation, in most cases leading rapidly to renal failure. The etiology and pathogenesis are only partly elucidated. Since irreversible renal scaring develops within days to weeks, RPGN represent a nephrological emergency necessitating urgent diagnostic evaluation and rapid institution of effective therapy. New onset nephritic sediment combined with concomitantly deteriorating excretory renal function should lead to immediate nephrological consultation. Autoimmune serology and particularly renal biopsy are of the utmost importance for rapid diagnosis. Most forms of RPGN are treated with immunosuppressive regimens which generally consist of high dose steroids in combination with the alkylating agent cyclophosphamide. Some forms also require the use of plasma exchange therapy. Rapid diagnosis and early therapy improves both renal and overall outcome in the affected patients. PMID- 14566465 TI - [Membranous glomerulonephritis]. AB - Membranous nephropathy remains the most common cause of nephrotic syndrome in adults. The common variant is idiopathic membranous nephropathy with no evidence of any known precipitating factors. Membranous nephropathy also occurs as a secondary form in association with inflammatory or neoplastic diseases. Prognosis is mostly favorable as shown by the frequency of spontaneous remissions which averages 30%, although about one-third of patients progress to end-stage renal failure. Risk factors for a poor prognosis include severe proteinuria, hypertension, older age, male gender and impaired renal function. Therapy should include an ACE-Inhibitor and/or angiotensin-II receptor blocker to lower proteinuria (blood pressure < or =130/80 mmHg). The majority of patients should be observed for six months whilst receiving conservative treatment before deciding about an immunosuppressive approach. The debate over its management continues today. Steroids alone are ineffective. Evidence-based medicine supports the use of cyclosporine or the Ponticelli regimen (monthly cycling routine of chlorambucil or cyclophosphamide alternating with prednisone). PMID- 14566467 TI - [Pulmonary renal syndrome]. AB - Pulmonary-renal syndrome is a potentially life-threatening disorder, characterised by diffuse alveolar hemorrhage on the basis of pulmonary capillaritis in association with rapidly progressive glomerulonephritis. It can originate from various systemic autoimmune diseases. ANCA-associated vasculitides account for approximately 60% and Goodpasture's syndrome for approximately 20% of the cases. Fulminant pulmonary capillaritis can result in acute respiratory failure with the more subtle forms only being detectable by bronchoalveolar lavage. Kidney biopsy displays extracapillary proliferating glomerulonephritis and renal immunohistology facilitates detection of the underlying systemic disease. By accelerating the diagnosis of the specific underlying disease, auto antibody testing fosters rapid initiation of treatment and thereby strongly improves the prognosis of pulmonary-renal syndrome. Intense immunosuppression with cyclophosphamide and glucocorticoids, augmented by plasmapheresis in the event of Goodpastures's syndrome, is the mainstay of therapy. Supportive measures such as temporary ventilation and hemodialysis have further reduced mortality. PMID- 14566466 TI - [IgA nephropathy: frequent, but rarely diagnosed]. AB - IgA nephropathy (IgAN) is the most common type of glomerulonephritis in the western world. In the majority of cases, it manifests in adolescence or early adulthood as recurrent macrohematuria, frequently triggered by infections, or persistent microhematuria as well as mild proteinuria, hypertension and/or renal insufficiency. In view of the later, it is not surprising that IgAN is often a chance finding. The majority of affected persons probably never come to medical attention, since in autopsies a prevalence of up to 1% of the population has been reported. About 20-30% of patients with a diagnosis of IgAN suffer from chronic, slowly progressive renal failure. Predictors include the degree of proteinuria and arterial hypertension as well as the established renal impairment at the time of diagnosis. Early identification of this risk group is of particular importance, since adequate therapy can stop or at least retard the progression of renal failure. When end stage renal failure has developed and a renal transplant is performed, about 25% of the patients will experience a clinically relevant recurrence of IgAN with progressive graft dysfunction. PMID- 14566468 TI - [Atypical resting ecg pattern in a patient with a positive family history of sudden cardiac death]]. AB - A 42 year old male patient presented with atypical ECG pattern at rest and reported that his brother died suddenly of unknown reasons at the age of 40. A performed pharmacological testing with ajmalin discovered strong ST-segment elevations in lead V(1) and V(2) in combination with an incomplete right bundle branch block. As a result, the diagnosis Brugada syndrome was established. The Brugada syndrome is a primary cardiac conduction disturbance without structural heart disease and is associated with an increased risk of sudden cardiac death caused by life threatening ventricular tachyarrhythmia. It is an inherited disease displaying an autosomal dominant mode of transmission with an incomplete penetrance, especially prevalent in females. In some regions like the southeast of Asia and Japan it is endemic. The mutation is linked to the sodium channel gene SCN5A. The annual mortality rate is estimated 30 per 100.000 persons. Today, exact diagnostic criteria do not exist. The diagnosis is based on the typical ECG pattern, combined with clinical symptoms, and a family history of sudden cardiac death. The currently suggested therapy consists of the implantation of an internal defibrillator. PMID- 14566469 TI - [A 41 year-old male patient with fever of unknown origin and bacteremia with actinobacillus actinomycetemcomitans]. AB - A 41-year-old male patient presented with fever of unknown origin which had been present for 5 months. The primary diagnostic procedures did not identify an infectious focus. After a lag phase of 13 days, blood cultures became positive for Actinobacillus actinomycetemcomitans, which belongs to the HACEK group of microorganisms. According to the DUKE criteria, infective endocarditis was diagnosed despite negative transesophageal echocardiography (TEE). The sensitivity of TEE is estimated between 86 and 94%. An infected tooth was extracted as a possible focus, and the patient was treated with i.v. antibiotics for 5 weeks according to the guidelines of the AHA and ACC. The fever was permanently terminated, and the further course of the patient was uneventful. Despite negative TEE, the diagnosis "infective endocarditis" should not be rejected, and blood cultures should be grown for up to 30 days. PMID- 14566470 TI - [Drug treatment of sleep disorders in the elderly]. AB - Changes in the pattern of sleep, e.g. more frequent nocturnal awakenings, are normal in the elderly. There is also a greater incidence of medical and psychiatric sleep disorders (depression, dementia). Initially, a thorough diagnostic assessment should be performed in order to identify disorders which can be treated specifically. For the symptomatic drug treatment of insomnias, the specific metabolic and pharmacokinetic, as well as possible interactions, should be considered. The new benzodiazepine receptor agonists (zopiclone, zolpidem and zaleplone), with their favourable risk-benefit profile, can be considered as first-choice treatments in elderly patients; in general, they should be preferred to the classical benzodiazepines. When a longer treatment is necessary, certain (non-anticholinergic) antidepressants and neuroleptics can be considered (the latter especially in cases of abnormal nocturnal behaviour). Herbal drugs and other hypnotically active compounds play a secondary role. Drug treatment of insomnia should always be carried out in the context of a general treatment plan which also includes non-pharmacological elements. In elderly patients, "chronotherapeutic" methods, which accentuate the sleep-wake rhythm, are of crucial importance. PMID- 14566471 TI - [The ALLHAT study (antihypertensive and lipid-lowering treatment to prevent heart attack trial): primary antihypertensive agents - diuretics vs calcium channel blockers vs angiotensin-converting enzyme inhibitors]. PMID- 14566473 TI - Can the addition of regional radiotherapy counterbalance important risk factors in breast cancer patients with extracapsular invasion of axillary lymph node metastases? AB - PURPOSE: To evaluate if locoregional radiotherapy (RT) versus local irradiation only can alter the pattern of failure in breast cancer patients with extranodal invasion. PATIENTS AND METHODS: From 08/1988 to 06/1998, 81 patients with extranodal invasion were treated with adjuvant RT (median total dose: 50.4 Gy), 46/81 only locally, 35/81 loco regionally due to presumed adverse parameters. The mean number of resected (positive) lymph nodes was 17 (seven). 78 patients received adjuvant systemic treatment(s). RESULTS: Patients treated with locoregional RT had significantly more often lymphatic vessel invasion (LVI; 63% vs. 28%; p = 0.003), T3/T4 tumors (43% vs. 17%; p = 0.014), and four or more positive lymph nodes (91% vs. 46%; p < 0.001) than patients irradiated only locally. Disease progression occurred in 24/81 patients (locoregional RT: 26% vs. local RT: 33%). The above risk factors were highly significant of worse outcome. Despite their overrepresentation in the locoregional RT group, no difference was found between both groups in regard to disease-free survival (DFS; p = 0.83) and overall survival (OS; p = 0.56), suggesting that regional RT was able to counterbalance the increased risk. There was even a trend toward a better 3-year DFS, 61% in locoregional RT and 37% in local RT, in the subgroup of patients with four or more positive lymph nodes. In a Cox regression model, higher T-stage, four or more positive lymph nodes, and LVI remained significant. For DFS and distant metastasis-free survival (DMFS), the absence of estrogen receptors and the omission of regional RT were also significant. CONCLUSION: Our data suggest that the addition of regional RT might be beneficial in selected subgroups of patients with extranodal invasion and other poor prognostic factors. PMID- 14566474 TI - [Radiation-induced mucositis and neutrophil granulocytes in oral mucosa]. AB - BACKGROUND: Chemotherapy-induced mucositis can be related to a decrease in oral neutrophils. We tested the relationship between radiation-induced mucositis and oral neutrophil counts. PATIENTS AND METHODS: Oral neutrophil counts were obtained for ten patients with head and neck cancer who received radiotherapy of the pharynx and oral cavity. Four patients received additional chemotherapy (5 FU, Mitomycin). Counts were obtained before and during treatment; four healthy volunteers were included in the study as well. For evaluation, a quantitative mouth rinse assay, including neutrophil-staining with acridin-orange, was applied. RESULTS: We observed large inter-individual variations with respect to neutrophil counts for patients and control persons (Table 1). During treatment (irradiation or chemoirradiation), large intra-individual variations were seen additionally (Figure 1). We found a correlation between neutrophil counts and clinical reaction grade. Neutrophil counts increased with increasing mucositis (Figure 2). This increase was more pronounced for patients treated with chemoirradiation compared to radiation alone. Treatment breaks at weekends had no clear influence on neutrophil counts. CONCLUSIONS: We observed a weak correlation between neutrophil counts and clinical reaction grade. However, the variations in neutrophil counts are too large to utilize this parameter as a surrogate for clinical mucositis grading. The assumption that a decrease in oral neutrophils is associated with radiation-induced mucositis was clearly negated. PMID- 14566475 TI - Aggressive simultaneous radiochemotherapy with cisplatin and paclitaxel in combination with accelerated hyperfractionated radiotherapy in locally advanced head and neck tumors. Results of a phase I-II trial. AB - BACKGROUND: Simultaneous radiochemotherapy (sRCT) is the treatment of first choice in locally advanced head and neck cancers. We have tested a very aggressive combination protocol with cisplatin and escalated paclitaxel in combination with accelerated hyperfractionated radiotherapy to assess the maximum tolerated dose (MTD), dose-limiting toxicity (DLT), overall toxicity, and response rate. PATIENTS AND METHODS: The trial recruited 24 patients (21 males, three females, mean age 57 years) treated at our department from 1998 through 2001. Irradiation was administered in daily doses of 2 Gy up to 30 Gy followed by 1.4 Gy twice daily up to 70.6 Gy to the primary tumor and involved nodes and 51 Gy to the clinically negative regional nodes. The chemotherapy schedule included cisplatin in a fixed dose of 20 mg/m(2) on days 1-5 and 29-33 and paclitaxel at increasing dose levels of 20, 25, 30 mg/m(2) twice weekly over the whole treatment time. Patients were recruited in cohorts of three to six, and the MTD was reached if two out of six patients in one cohort developed DLT. DLT was defined as any grade 4 toxicity or any grade 3 toxicity requiring treatment interruption or unplanned hospitalization or any grade 3 neurotoxicity. We recruited mainly patients with large tumors for this protocol; all patients were stage IV, and the mean tumor volume (primary + metastases) amounted to 72 +/- 61 cm(3). The mean follow-up was 30 months (range 4-39 months). RESULTS: One early death (peritonitis and sepsis at day 10) occurred, and 23 patients were evaluable for acute toxicity and response. The MTD of paclitaxel was reached at the third dose level (30 mg/m(2) paclitaxel twice weekly). The DLT was severe mucositis grade 3 (n = 1) and skin erythema grade 4 (n = 2). After determining the MTD, another 14 patients were treated at the recommended dose level of paclitaxel with 25 mg/m(2) twice weekly. In summary, 13/23 patients (57%) developed grade 3 and 10/23 (43%) grade 2 mucositis. Two patients (9%) had grade 4, five (22%) grade 3, and 16 (69%) grade 2 dermatitis. One patient died at day 30 of neutropenic infection. In one patient, a grade 2 nephrotoxicity appeared requiring cessation of cisplatin chemotherapy. 18/23 patients (78%) required blood transfusion (1-3 units) and 16/23 (70%) i.v. antibiotics. 14 patients (61%) achieved a complete and nine (39%) a partial remission, yielding an overall response rate of 100%. In summary, six patients died of local tumor progression (n = 2), distant metastases (n = 2), or therapy-related complications (n = 2) during follow-up. The 3-year overall survival was 71%. Tumor volume was not a risk factor for failure in this protocol (mean tumor volume in relapse-free vs. progressive patients 71 +/- 65 cm(3) vs. 64 +/- 38 cm(3)). All patients have, so far, developed only slight late effects (fibrosis, lymphedema) with no grade 3-4 late sequelae. CONCLUSIONS: This very aggressive sRCT protocol yielded excellent response and survival figures but was associated with a very high rate of acute toxicity (8% therapy-related deaths). A maximal supportive treatment is therefore required. PMID- 14566476 TI - [Evaluation of quality of life of patients with oral squamous cell carcinoma. Comparison of two treatment protocols in a prospective study-first results]. AB - BACKGROUND: The treatment of oral cancer has a strong impact on the quality of life. In recent years different therapeutic concepts have been developed, these include preoperative simultaneous "neoadjuvant" radiochemotherapy (RCT) and one stage surgery with tumor ablation and reconstruction. When considering long-term survival, there is substantial evidence that evidenced modality treatment including neoadjuvant RCT is superior to the primary surgical approach with postoperative radiation. PATIENTS AND METHODS: This longitudinal study prospectively evaluates quality of life in two groups consisting of 53 neoadjuvant and primarily surgically treated patients with oral cancer, using the quality-of-life core questionnaire (QLQ-C30) and the head and neck cancer module (H and N 35) of the European Organization for Research and Treatment of Cancer (EORTC). RESULTS: Postoperatively both groups showed a marked reduction in quality of life. 1 year later quality of life had equalized between the two groups to such an extent that the quality of life scores had almost reached the preoperative level. Both groups showed specific impairments in the symptom scales. In the neoadjuvant therapy group however, global health and the emotional status were reduced to a greater degree than in the other group. CONCLUSION: Temporary limitations in quality of life can be expected after tumor treatment of oral cancer as presented here. Neoadjuvant therapy concept is more aggressive and might result in a longer disease-free survival, but the restriction in quality of life is more severe. Primary goal is the eradication of the tumor. Nevertheless preservation or reconstruction of a maximum of function is essential for a high level of quality of life. PMID- 14566477 TI - Increased metabolic activity in the spinal cord of patients with long-standing Lhermitte's sign. AB - PURPOSE: To investigate the pathophysiology of the radiation-induced, chronic Lhermitte's sign (LS) on the basis of long-standing case histories with partial functional recovery. PATIENTS AND METHODS: As radiotherapy in two nasopharyngeal cancer patients, a biologically effective dose (BED) of 103.8 Gy(2) (case 1) and 94.8 Gy(2) (case 2) was delivered to the cervical spinal cord. Neurologic signs relating to the irradiated spinal cord segments developed after 2 months (case 1) and 5 years (case 2), with radiation-induced damage equivalent to grade 3 (case 1) and grade 2 (case 2) toxicity (Common Toxicity Criteria, Version 2.0). The clinical status improved to grade 2 (case 1) and grade 1 (case 2). Positron emission tomography (PET) and fibroblast clonogen assay were applied 25 and 7 years postirradiation, respectively, to characterize this rare clinical picture. RESULTS: PET demonstrated increased [(18)F]fluorodeoxyglucose (FDG) accumulation and [(15)O]butanol perfusion, but negligible [(11)C]methionine uptake in the irradiated spinal cord segments in both patients. In clonogenic assays, fibroblasts from case 1 displayed much higher radiation sensitivity than in healthy controls, while in case 2 the fibroblasts sensitivity was normal. CONCLUSIONS: These data suggests a close direct relationship between regional perfusion and metabolism of the spinal cord, similarly as in the brain. The postirradiation recovery may be related to energy-demanding conduction, explaining the increased metabolism and perfusion. The increased radiosensitivity and higher spinal cord BED may have contributed to the more severe sequelae in case 1. PMID- 14566478 TI - Radiotherapy for stage I-III nodal low-grade non-Hodgkin's lymphoma. AB - BACKGROUND: To evaluate retrospectively long-term results and patterns of recurrence in patients with low-grade non-Hodgkin's lymphoma (NHL) Ann Arbor stage I-II and limited stage III. PATIENTS AND METHODS: 58 patients, who had been treated between 1980 and 1996, were analyzed. Median follow-up period was 8.75 years. 48 patients (83%) presented with follicular lymphoma (stage I: 23 patients, stage II and III: 15 and ten patients, respectively). Median age was 51 years. Irradiation was applied with a median total dose of 40 Gy. 13 patients (22%) additionally received chemotherapy. End points of the investigation were remission rate, overall- and disease-free survival, and patterns of recurrence, as well as the prognostic impact of age, B-symptoms, chemotherapy, irradiation dose, treatment volume, and Ann Arbor stage. RESULTS: 6 weeks after treatment 91% of the patients had complete, 7% partial response. One patient (2%) was classified as progressive disease. Overall survival rate was 86% and 69% at 5 and 10 years, respectively. Corresponding disease-free survival rates were 73% and 63%. Regarding overall survival, multivariate analysis identified age (p = 0.001) as independent prognostic factor. In the subgroup of patients with follicular lymphoma 92% were found in complete, 6% in partial remission, one patient (2%) with progressive disease. Overall survival rates at 5 und 10 years were 87% and 70%, disease-free survival rates 75% and 64%, respectively. Out-of-field recurrence rate for all 58 patients was 34% and the proportion of relapses at nodal or lymphatic sites outside the treated areas in relation to all registered recurrences was 77%. CONCLUSIONS: Our results maintain external radiotherapy as a curative concept in the treatment of limited stage low-grade lymphoma, especially in younger patients. Patterns of recurrence would favor total nodal irradiation (TNI) as an appropriate approach for these patients. PMID- 14566479 TI - Enhanced renal toxicity of total body irradiation combined with radioimmunotherapy. AB - PURPOSE: Total body irradiation (TBI) with and without additional radioimmunotherapy (RIT) was examined for renal toxicity after stem cell transplantation. PATIENTS AND METHODS: Serum creatinine levels of 35 patients (15 female, 20 male, median age 40.5 years, range 17-60 years) after TBI alone and of 23 patients (eight female, 15 male, median age 47, range 16-58 years) after TBI with additional RIT were determined between 10/1997 and 11/1999. TBI was performed by external-beam radiotherapy in six fractions over 3 days with renal doses of 12 Gy in the TBI-alone group and 6 Gy in the group with additional RIT. The mean kidney dose due to the (188)Re-radiolabeled antibody was estimated to be 8.3 Gy (2.3-11.6 Gy). RESULTS: Within 12 months after treatment, creatinine levels increased from 77 mmol/l (SD +/- 11) to 89 mmol/l (SD +/- 20) for TBI alone and from 78 mmol/l (SD +/- 13) to 144 mmol/l (SD +/- 52) for combined TBI and RIT. CONCLUSION: Despite a 50% reduction of the external-beam contribution to the kidney dose, the application of approximately 10 GBq (188)Re-labeled anti CD66 monoclonal antibody with a calculated renal dose of 8.3 Gy (range 2.3-11.5 Gy) led to renal toxicity, as reported previously. In the absence of a positive dose-response relationship for the (188)Re-labeled antibody, the observation may be explained by an underestimation of the biologically effective dose and the inaccuracy of the dose determination at the glomerular level. PMID- 14566480 TI - Moist skin care can diminish acute radiation-induced skin toxicity. AB - BACKGROUND: Radiation treatment may induce acute skin reactions. There are several methods of managing them. Validity of these methods, however, is not sufficiently studied. We therefore investigated, whether moist skin care with 3% urea lotion will reduce acute radiation skin toxicity. PATIENTS AND METHODS: 88 patients with carcinomas of the head and neck undergoing radiotherapy with curative intent (mean total dose 60 Gy, range: 50-74 Gy) were evaluated weekly for acute skin reactions according to the RTOG-CTC score. In 63 patients, moist skin care with 3% urea lotion was performed. The control group consisted of 25 patients receiving conventional dry skin care. The incidence of grade I, II, and III reactions and the radiation dose at occurrence of a particular reaction were determined and statistically analyzed using the log-rank test. The dose-time relations of individual skin reactions are described. RESULTS: At some point of time during radiotherapy, all patients suffered from acute skin reactions grade I, > 90% from grade II reactions. 50% of patients receiving moist skin care experienced grade I reactions at 26 Gy as compared to 22 Gy in control patients (p = 0.03). Grade II reactions occurred at 51 Gy versus 34 Gy (p = 0.006). Further, 22% of the patients treated with moist skin care suffered from acute skin toxicity grade III as compared to 56% of the controls (p = 0.0007). CONCLUSION: Moist skin care with 3% urea lotion delays the occurrence and reduces the grade of acute skin reactions in percutaneously irradiated patients with head and neck tumors. PMID- 14566481 TI - [Staffing levels in medical radiation physics in radiation therapy in Germany. Summary of a questionnaire]. AB - To get a general idea of the actual staffing level situation in medical radiation physics in 1999 a survey was carried out by the task-group "Personalbedarf" of Deutsche Gesellschaft fur Medizinische Physik (DGMP) among all DGMP-members who are active in this field. Main components for equipment and activities are defined in Report 8 and 10 of DGMP for staffing requirements in medical radiation physics. 322 forms were sent out, 173 of them have been evaluated. From the answers regarding equipment and activities numbers for staff are calculated by the methods given in Report 8 and 10 for this spot check target and compared with effective staffing levels. The data of the spot check are then extrapolated on total Germany. The result is a calculated deficit of 865 medical physicists for the whole physics staff, 166 of them in radiation therapy. From the age distribution of DGMP-members and the calculated deficit resulted a training capacity of about 100 medical physicists at all per year (19 in radiation therapy) if the deficit shall be cut back in 10 years. PMID- 14566482 TI - A family with spinal anaplastic ependymoma: evidence of loss of chromosome 22q in tumor. AB - Familial ependymal tumors are a very rare disease, the pathogenesis of which is unknown. Previous studies indicate an involvement of tumor suppressor genes localized within chromosomal region 22q, whereas details are still unclear. Here we report a non-neurofibromatosis type-2 (non-NF2) Japanese family in which two of the four members are affected with cervical spinal cord ependymoma, and one of the four is affected with schwannoma. Loss of heterozygosity (LOH) studies were carried out searching for common allelic loss at chromosomal region 22q11.2-qtel in two of the affected patients. Our findings support a prediction for existence of a tumor suppressor gene on chromosome 22 especially related to the tumorigenesis of familial ependymal tumors. PMID- 14566483 TI - Different attenuated phenotypes of GM2 gangliosidosis variant B in Japanese patients with HEXA mutations at codon 499, and five novel mutations responsible for infantile acute form. AB - Eight mutations of the alpha subunit of beta-hexosaminidase A gene ( HEXA) were identified in eight patients with GM2 gangliosidosis variant B. They were five missense mutations, two splice-site mutations, and one two-base deletion. Five of them, R252L (CGT-->CTT), N295S (AAT-->AAC), W420C (TGG-->TGT), IVS 13, +2A-->C, and del 265-266AC (exon 2), were novel mutations responsible for infantile acute form of GM2 gangliosidosis. Two missense mutations, R499H and R499C, were found in one allele of two patients with attenuated phenotypes. The patient with R499C showed a late infantile form, and the other patient with R499H showed a juvenile form. These two mutations have been reported previously in the patients of other ethnic groups, and they have been known to cause attenuated phenotypes. The milder phenotypes of GM2 gangliosidosis variant B, different from the infantile acute form, have not been reported so far in Japan, and this is the first report of Japanese patients with attenuated phenotypes and their molecular analysis. PMID- 14566484 TI - Identification and characterization of a novel splice-site mutation in a patient with Wiskott-Aldrich syndrome. AB - Wiskott-Aldrich syndrome (WAS) is an X-linked recessive disorder characterized by immunodeficiency, eczema, and thrombocytopenia with small platelets. A wide spectrum of mutations in the Wiskott-Aldrich syndrome protein ( WASP) gene have been identified as causative of the disease. In the present paper, we report on a family with a boy affected by WAS, with a splice-site mutation caused by a T to G substitution in the +2 position of intron 6 (IVS6+2T>G). Expression studies performed in COS-7 and U-937 cells showed that the mutation affected the normal splicing process. As a consequence, an abnormally long transcript of 38 nucleotides is generated. Such missplicing is probably due to the activation of a cryptic splice donor site located 38 nt downstream of exon 6. The translation of such aberrant mRNA will produce a truncated protein with a premature stop at codon 190. Thus, a novel splice-site mutation is reported in a patient with a mild WAS phenotype. PMID- 14566487 TI - The effect of thoracic sympathectomy on baroreflex control of circulation. PMID- 14566485 TI - Relationships between soil heavy metal concentration and mycorrhizal colonisation in Thymus polytrichus in northern England. AB - A study was conducted to establish whether the wild thyme [ Thymus polytrichus A. Kerner ex Borbas ssp. britannicus (Ronn.) Kerguelen (Lamiaceae)] growing in the metal-contaminated soils along the River South Tyne, United Kingdom, is colonised by arbuscular mycorrhizal (AM) fungi, and whether the degree of colonisation increases (perhaps suggesting increasing mycorrhizal dependence) or decreases (indicating possible inhibition of AM growth) with increasing degree of soil contamination. Seasonal changes in AM colonisation were also assessed. The AM fungal communities colonising T. polytrichus were also investigated, using the polymerase chain reaction with restriction fragment length polymorphism and sequencing of fungal DNA to establish whether AM species richness varied between sites, and whether fungal ecotypes specific to sites with different amounts of metal contamination could be identified. All plants examined were heavily colonised by AM fungi, and mean percentage root length colonised did not increase significantly with increasing soil metal contamination. However, AM vesicle abundance (percentage of mycorrhizal root length containing vesicles) at the most contaminated site was significantly greater than at the other sites. No significant seasonal variation in degree of colonisation or vesicle abundance was found. Glomus was the predominant AM genus detected at all sites. The number of AM genotypes colonising T. polytrichus roots was similar at all sites but, although some were common to all sites, certain strains appeared to be specific to either the most- or the least-contaminated site. This variation in species may account for the difference in vesicle abundance between sites. The consistently heavy AM colonisation of T. polytrichus found suggests that these fungi are not inhibited by soil heavy metals at these sites, and that the host derives some benefit from its AM symbiont. PMID- 14566488 TI - Endoscopic thoracic sympathectomy attenuates reflex tachycardia during head-up tilt in lightly anesthetized patients with essential plamar hyperhidrosis. AB - PURPOSE: Our purpose was to examine perioperative alterations in hemodynamic changes with head-up tilt (HUT) in patients undergoing endoscopic thoracic sympathectomy (ETS). METHODS: The subjects were 11 patients with essential hyperhidrosis scheduled to undergo ETS (ETS group) and 9 age-matched volunteers undergoing minor surgery (control group). HUT was performed (40 degrees; 5 min) before and after the surgery, under nitrous oxide anesthesia. Orthostatic hypertension and hypotension in response to HUT were defined as changes of 10% or greater in systolic blood pressure. RESULTS: The increase in heart rate in response to HUT was significantly reduced after surgery in the ETS group (from 34 +/- 18 to 14 +/- 11 beats.min(-1); P < 0.001), but not in the control group (from 23 +/- 18 to 22 +/- 12 beats.min(-1); P = 0.911). Orthostatic hypertension disappeared completely after ETS (from 5 of 11 to none of 11 patients; P = 0.035), whereas the prevalence of orthostatic hypotension increased significantly after ETS (from 3 of 11 to 9 of 11 patients; P = 0.030). In the control group, the prevalence of neither orthostatic hypertension nor orthostatic hypotension changed after surgery. CONCLUSIONS: ETS attenuates autonomic circulatory response under nitrous oxide anesthesia. PMID- 14566489 TI - Flumazenil reduces the hypnotic dose of propofol in male patients under spinal anesthesia. AB - PURPOSE: Flumazenil has been reported to produce a partial benzodiazepine-agonist like effect in some psychopharmacological examinations. This study investigated the effect of flumazenil on the hypnotic activity of propofol in 60 men scheduled for minor surgical procedures done under spinal anesthesia. METHODS: After a steady state of spinal anesthesia had been reached, patients were pretreated with saline or flumazenil, 5 microg.kg(-1), followed by the administration of saline or midazolam, 10 microg.kg(-1). Then, 250 microg.kg(-1).min(-1) of propofol was infused until hypnosis was achieved. Loss of response to a simple command with a slight stimulus, served as the end-point for hypnosis. Immediately after achievement of the end-point, propofol infusion was discontinued, and a 2-ml venous blood sample was obtained from the dorsal pedis vein to determine plasma propofol concentration. RESULTS: Flumazenil significantly decreased the dose of propofol required and the time required to achieve hypnosis compared with values in the control group (55 +/- 10 [mean +/- SD] vs 71 +/- 14 mg and 212 +/- 42 vs 268 +/- 48 s, respectively; P < 0.05), whereas flumazenil attenuated the effect of midazolam in reducing the plasma concentration of propofol at hypnosis (2.9 +/ 0.5 and 2.5 +/- 0.6 microg.ml(-1), respectively; P < 0.05). CONCLUSION: These results suggested that flumazenil may potentiate the hypnotic properties of propofol, despite flumazenil having an antagonistic effect on the enhanced hypnotic activity of propofol induced by the coadministration of midazolam. PMID- 14566490 TI - Thoracoscopic sympathectomy: endobronchial anesthesia vs endotracheal anesthesia with intrathoracic CO2 insufflation. AB - PURPOSE: To compare clinical advantages and hemodynamic and respiratory changes during one lung-collapsed ventilation (OLCV) using a double-lumen tube (DLT) or a single-lumen tube (SLT) with intrathoracic CO(2) insufflation, in patients undergoing thoracic sympathectomy (TS) under general anesthesia. METHODS: One hundred and twenty-five patients (94 men and 31 women) undergoing TS for the treatment of palmar hyperhidrosis (PH) were randomly allocated to two groups: group A (68 patients; age, 29 +/- 6 years) in whom DLT was used, and group B (57 patients; age, 32 +/- 3 years) in whom SLT with intrathoracic CO(2) insufflation at a rate of 0.5-1 l.min(-1) and sustained intrathoracic pressure at 6 mmHg insufflation were used. Anesthesia was maintained with 1 minimum alveolar concentration (MAC) isoflurane in 50% nitrous oxide in oxygen with incremental doses of sufentanil and atracurium when required. Arterial blood gases were measured in 10 patients in group B. Hemodynamic and respiratory parameters were obtained perioperatively. RESULTS: There were no significant differences in hemodynamic and respiratory parameters between the two groups during the study phases, except for the arterial oxygen saturation (SpO(2)). The times required for anesthesia and surgery were significantly shorter in the SLT group than in the DLT group. SpO(2) during OLCV was 95 +/- 1% with DLT and 98 +/- 1% with SLT, with a significant difference. Three patients had an SpO(2) of less than 90% in the recovery room, where the chest tube position was readjusted, with no further sequelae. CONCLUSION: General anesthesia with SLT and intrathoracic CO(2) insufflation provides optimal operating conditions, adequate oxygenation, and perfect hemodynamic stability during TS. PMID- 14566491 TI - Renal function in surgical patients after administration of low-flow sevoflurane and amikacin. AB - PURPOSE: Compound A, a degradation product of sevoflurane, is nephrotoxic in rats, while aminoglycosides induce nephrotoxic injury in humans. Combining an aminoglycoside with a known nephrotoxin can enhance nephrotoxicity. We investigated the effects of aminoglycosides on renal function in surgical patients anesthetized with low-flow sevoflurane. METHODS: We compared the urinary excretion of several biochemical markers (such as total protein, albumin, beta(2) microglobulin, glucose, and N-acetyl-beta-glucosaminidase [NAG]) in an amikacin group ( n = 18) and a control group ( n = 19) of surgical patients anesthetized with low-flow anesthesia (1 l.min(-1)) with sevoflurane. All patients received cefotiam as an antibiotic perioperatively. In addition, the amikacin group received amikacin, an aminoglycoside, given intravenously twice a day (400 mg per day) from immediately after the induction of anesthesia to day 2 after anesthesia. RESULTS: Duration of anesthesia and mean compound A concentration were 5.2 +/- 1.4 h and 27.2 +/- 8.7 ppm (mean +/- SD) in the amikacin group, and 5.1 +/- 1.7 h and 27.1 +/- 7.8 ppm in the control group respectively ( P > 0.05). The two groups did not differ in clinical laboratory baseline values (blood urea nitrogen and serum creatinine concentration). There were no significant differences between the groups in either the maximum or the average values for the urinary excretion of biochemical markers after anesthesia. CONCLUSION: Our study demonstrates that there is no synergic effect of compound A and amikacin on nephrotoxicity in humans. PMID- 14566492 TI - Comparison of heart rate changes after neostigmine-atropine administration during recovery from propofol-N2O and isoflurane-N2O anesthesia. AB - PURPOSE: Propofol augments the reduction of heart rate (HR) in combination with cholinergic agents and attenuates the HR response to atropine. We examined whether propofol anesthesia was associated with an increased incidence and extent of bradycardia after neostigmine-atropine administration compared with the effects of isoflurane anesthesia. METHODS: Thirty-six adult patients were randomly assigned to two groups ( n = 18 each): the propofol group patients were anesthetized with propofol (5-10 mg.kg(-1).h(-1))-(2)O-fentanyl, and the isoflurane group patients were anesthetized with isoflurane (0.5%-1.0%)-(2)O fentanyl. When surgery was completed, anesthetics were discontinued, and then a mixture of neostigmine 0.05 mg.kg(-1) and atropine 0.02 mg.kg(-1) was injected intravenously over 20 s. Blood pressure (BP) and HR were measured noninvasively at 1-min intervals for 10 min. RESULTS: At the completion of the surgery, the average infusion rate of propofol was 6.2 +/- 1.7 mg.kg(-1).h(-1), and the average inspired concentration of isoflurane was 0.73 +/- 0.15%. Immediately before the neostigmine-atropine injections, HR and mean BP were similar in the two groups. The maximum increase in HR after the neostigmine-atropine injections was significantly less in the propofol group than in the isoflurane group (16 +/- 9 and 34 +/- 6 beats.min(-1), respectively, P < 0.01). The subsequent maximum decrease in HR was greater in the propofol group than in the isoflurane group (-9 +/- 4 and -5 +/- 4 beats.min(-1), respectively; P < 0.01). The incidence of bradycardia (HR < 50 beats.min(-1)) after neostigmine-atropine injection was greater in the propofol group than in the isoflurane group (61% and 28%, respectively; P < 0.01). CONCLUSION: We conclude that propofol anesthesia attenuates the initial increases in HR, enhances the subsequent decreases in HR, and increases the incidence of bradycardia after neostigmine-atropine injections compared with the effects of isoflurane anesthesia. PMID- 14566493 TI - The effect of propofol infusion on minimum alveolar concentration of sevoflurane for smooth tracheal intubation. AB - PURPOSE: This study was conducted to determine the effect of propofol infusion on the minimum alveolar concentration necessary for smooth tracheal intubation (MACEI) of sevoflurane. METHODS: Sixty-nine patients, American Society of Anesthesiologists (ASA) status I, aged 30-49 years, were randomly assigned to one of three groups according to the agents used for tracheal intubation (n = 23 for each group): the SP group, in whom the intubation was attempted under sevoflurane plus propofol infusion; the S group, tracheal intubation under sevoflurane alone; and the P group, tracheal intubation under propofol infusion alone. Anesthesia was induced with propofol 2.5 mg.kg(-1) i.v. bolus. Prior to the tracheal intubation attempt, propofol infusion, 10 mg.kg(-1).h(-1), was given for 15 min in the SP and P groups, and sevoflurane equilibration was established in the SP and S groups. All tracheal intubation attempts were made 15 min after anesthetic induction. The end-tidal sevoflurane concentration at which tracheal intubation was attempted was predetermined by the up-and-down method (with 0.5% as a step size). MACEI was determined using a logistic regression test. RESULTS: The MACEI of sevoflurane was 1.73% in the SP group, and 2.99% in the S group. Laryngoscopy was not possible in the P group patients. CONCLUSION: Propofol infusion reduced sevoflurane MACEI. This finding suggests that propofol would be an excellent adjuvant to use with sevoflurane for tracheal intubation. PMID- 14566494 TI - Anticonvulsant effects of sevoflurane on amygdaloid kindling and bicuculline induced seizures in cats: comparison with isoflurane and halothane. AB - PURPOSE: We compared the anticonvulsant effects of sevoflurane with those of isoflurane and halothane in amygdaloid kindling and bicuculline-induced seizures in cats. METHODS: In a crossover design, the effects of 70% nitrous oxide, and 0.3, 0.6, and 1.5 minimum alveolar concentration (MAC) of volatile anesthetics were studied in five cats in which the amygdala was electrically stimulated at the current used for establishing the kindled state. The effects of 0.6 and 1.5 MAC of volatile anesthetics were studied in another five cats, in which 0.2 mg.kg(-1) of bicuculline was administered i.v.. RESULTS: In the amygdaloid kindling model, all four anesthetics decreased the duration of after-discharge (AD), the rise of multiunit activity in midbrain reticular formation (R-MUA), and the behavior scores compared with findings without anesthetics. Halothane, at 1.5 MAC, significantly decreased the number of cats showing AD ( P < 0.05). In the bicuculline-induced seizure model, all five cats showed repetitive spikes during 1.5 MAC of sevoflurane, whereas only two and three cats, respectively, showed the repetitive spikes during 1.5 MAC of isoflurane and halothane. All three volatile anesthetics decreased the rise of R-MUA, the duration of the repetitive spikes, and the behavior scores. The suppression of the rise in R-MUA and the behavior scores with 1.5 MAC of sevoflurane was significantly less than that with 1.5 MAC of isoflurane. CONCLUSION: The anticonvulsant effects of sevoflurane were less potent than those of halothane in the amygdaloid kindling model and less potent than those of isoflurane in the bicuculline-induced seizure model. PMID- 14566495 TI - Comparison between sevoflurane and isoflurane anesthesia in pig hepatic ischemia reperfusion injury. AB - PURPOSE: Sevoflurane and isoflurane have been reported to exert protective effects against ischemia-reperfusion injury (IRI) in various organs. To compare the effect of sevoflurane anesthesia on liver IRI with that of isoflurane anesthesia, we performed the present study in pigs. METHODS: Nineteen pigs were assigned to either the sevoflurane ( n = 9) or the isoflurane group ( n = 10). Hepatic warm ischemia was produced by 30-min hepatic artery and portal vein clamping beginning 90 min after the start of the inhalation anesthesia; this was followed by a 240-min reperfusion. To extend our evaluation, we evaluated the degree of IRI using various parameters (plasma alpha-glutathione-S-transferase [alpha-GST], lipid peroxide, and lactate concentrations), in addition to the conventionally used liver damage markers. RESULTS: The lactate level was significantly higher under isoflurane than under sevoflurane at 120 min after reperfusion (4.0 +/- 0.4 mmol.l(-1) vs 2.5 +/- 0.3 mmol.l(-1); P < 0.05). How ever, this difference had disappeared after 240 min of reperfusion. No significant differences between the two groups were observed in values for alpha GST, lipid peroxides, aspartate aminotransferase, alanine aminotransferase, or lactic dehydrogenase. CONCLUSION: The extent of the hepatic IRI seen under sevoflurane anesthesia in pigs did not differ significantly from that seen under isoflurane, as judged from measurements of a number of parameters over a 240-min reperfusion period. PMID- 14566496 TI - Anesthesia and the gastrointestinal tract. PMID- 14566497 TI - Education in anesthesiology for the twenty-first century. AB - The growth and development of anesthesiology in the twenty-first century will likely depend on two major factors: our vision for the specialty in the future and our ability to implement an anesthesia education plan that will foster the achievement of that vision. The foundation of effective anesthesia education must be built on an understanding of the past and an analysis of the present but, most importantly, it must be shaped by our vision for the future. Focus on the future is essential, for it is remarkably easy to teach others as we were taught, or as we practice today. Unfortunately, the easy path will not foster the advancement of the specialty or develop the leaders for the future. The comments that follow are not a prescription for success. Rather, they are intended to stimulate discussion and planning regarding the future of anesthesiology, leading to a course of action that will enhance the development of the specialty. Long-term success for the specialty will depend on our efforts in undergraduate and graduate medical education, whereas short-term success will depend on our efforts in the continuing medical education of current practitioners. PMID- 14566498 TI - Malignant hyperthermia with normal calcium-induced calcium release rate of sarcoplasmic reticulum in skeletal muscle. PMID- 14566499 TI - Accidental subarachnoid injection of atracurium: a case report. PMID- 14566500 TI - An adult with ARDS managed with high-frequency oscillatory ventilation and prone position. PMID- 14566501 TI - Blood component therapy guided by celite-activated thromboelastography for perioperative coagulopathy. PMID- 14566502 TI - Post-herpetic neuralgia in a patient with congenital insensitivity to pain and anhidrosis. PMID- 14566503 TI - Simple high-performance liquid chromatographic assay of propofol in human and rat plasma and various rat tissues. PMID- 14566504 TI - Unexpectedly severe hypoxia during sprint swimming. PMID- 14566505 TI - Perioperative coronary spasm reported in Japanese journals. PMID- 14566507 TI - Is prophylactic mechanical ventilation really necessary after esophageal surgery? PMID- 14566508 TI - Impairment of chest wall mechanics and increased chest wall work of breathing cause postoperative respiratory failure in patients who have undergone radical esophagectomy. AB - PURPOSE: We verified the hypothesis that impairment of chest wall mechanics would be related to the cause of postoperative respiratory failure in patients undergoing radical esophagectomy. METHODS: A total of 21 patients were studied. After management with mechanical ventilation to prevent respiratory failure for several days, trial weaning from the ventilator was performed. The patients were divided into a successful weaning group (S group) and an unsuccessful weaning group (US group), depending on the results of the weaning trial. We compared respiratory mechanics and the respiratory work of breathing during weaning from the ventilator between the two groups. RESULTS: In the US group, lung and chest wall compliance was significantly lower and the development of intrinsic positive end-expiratory pressure (PEEPi) was observed. The work of breathing and the oxygen cost of breathing were significantly higher in the US group. The increased respiratory energy work was due to a moderate increase in lung work and a significant increase in chest wall work. CONCLUSION: Our results suggested that postoperative respiratory failure was related to increased respiratory energy expenditure, significantly deteriorated chest wall mechanics, and increased chest wall work, all of which are involved in the development of postoperative respiratory failure after radical esophagectomy. PMID- 14566509 TI - The combination of a fiberoptic stylet and a McCoy laryngoscope facilitates tracheal intubation in difficult airway cases. AB - PURPOSE: Fiberoptic stylets are considered useful for difficult airway management. In the present study, we assessed the usefulness of a fiberoptic stylet when the stylet was used with a Macintosh or a McCoy laryngoscope. METHODS: Twenty-four patients, whose airways were graded as Cormack grade III, were studied. We compared the times required for tracheal intubation when the fiberoptic stylet was used with a Macintosh direct laryngoscope and when it was used with a McCoy laryngoscope. Cormack grade III was subdivided into IIIa (with distance between the epiglottis and the posterior wall of the pharynx) and IIIb (with no distance between the epiglottis and the posterior wall of the pharynx), according to the view of the vocal cords by the laryngoscope. RESULTS: The intubation time in grade IIIb patients, who were intubated by the concurrent use of the fiberoptic stylet and the McCoy laryngoscope (28 +/- 4 s), was not significantly different from that in grade IIIa patients (28 +/- 10 s). The intubation time in grade IIIb patients, who were intubated by the concurrent use of the fiberoptic stylet and the Macintosh laryngoscope (52 +/- 8 s), was significantly longer than that in grades IIIa (28 +/- 10 s; P < 0.01) or IIIb with the McCoy laryngoscope (28 +/- 4 s; P < 0.01). CONCLUSION: The combination of a new handy fiberoptic stylet and a McCoy laryngoscope facilitated tracheal intubation of patients whose airway had no distance between the epiglottis and the posterior wall of the pharynx in laryngoscopic vocal cord view. PMID- 14566510 TI - Effect of infraorbital nerve block under general anesthesia on consumption of isoflurane and postoperative pain in endoscopic endonasal maxillary sinus surgery. AB - PURPOSE: The efficacy of infraorbital nerve block in reducing isoflurane consumption and postoperative pain was evaluated in patients undergoing endoscopic endonasal maxillary sinus surgery (ESS) under general anesthesia. METHODS: Fifty patients were randomly allocated to either the block group (n =15) or the nonblock group (n = 25). After the establishment of general anesthesia with isoflurane, nitrous oxide, and oxygen, the patients received infraorbital nerve block with 1.0 ml of either 0.5% bupivacaine (block group) or normal saline (nonblock group) administered into the soft tissue in front of the infraorbital foramen. Systolic blood pressure during anesthesia and surgery was maintained at 85-90 mmHg by adjusting the inspiratory concentration of isoflurane, and its consumption was evaluated in both groups. Pain intensity at 15 min after the end of anesthesia was also evaluated on a five-point pain scale. RESULTS: The consumption of isoflurane under a fresh gas flow of 6 l.min(-1) was 17.3 +/- 6.5 ml.kg(-1).h(-1) (mean +/- SD) in the block group and 27.4 +/- 9.4 ml.kg(-1).h(-1) in the nonblock group during surgery ( P < 0.001). Nicardipine was required during surgery less frequently in the block group than in the nonblock group ( P < 0.01). Postoperative pain intensity was lower in the block group than in the nonblock group ( P < 0.01). CONCLUSION: General anesthesia combined with infraorbital nerve block is effective in reducing the consumption of isoflurane and postoperative pain intensity in ESS. PMID- 14566511 TI - Respiratory system compliance and postoperative ventilator dependence in neonates with left-sided congenital diaphragmatic hernia. AB - PURPOSE: The purpose of this prospective consecutive patient study was to test if perioperative respiratory system compliance of patients undergoing surgical repair of congenital diaphragmatic hernia (CDH) can predict the need for prolonged postoperative mechanical ventilation. METHODS: All neonates over 35 weeks of gestation who had surgical repair of left-sided CDH between July 1994 and December 1996 ( n = 10) were included in this study. Static respiratory system compliance (C(rs)) was measured by the passive occlusion method with muscle relaxation before (C(pre)) and after (C(post)) surgical repair of left sided CDH. We examined the relationship between respiratory system compliance and postoperative ventilator-dependent duration. RESULTS: The mean birthweight corrected C(post) was lower than that of birthweight-corrected C(pre) (0.41 +/- 0.18 vs 0.54 +/- 0.18 ml.cm H(2)O(-1).kg(-1), P = 0.04). One neonate died on postoperative day 31 from intraventricular hemorrhage, and the other nine neonates survived. The patient who died was excluded from the rest of our calculations. The ventilator-dependent duration was 16.7 +/- 12.3 days. The postoperative ventilator-dependent duration was longer when the birthweight corrected C(post) was smaller, with a significant correlation between these parameters ( P = 0.006). CONCLUSION: C(post), but not C(pre), may be useful to predict the need for prolonged postoperative mechanical ventilation in neonates with left-sided CDH. PMID- 14566512 TI - Do age-based formulae predict the appropriate endotracheal tube sizes in Japanese children? AB - PURPOSE: Age-based formulae have been widely used to predict the appropriate size of the endotracheal tube (ETT) in children. These formulae are based on old data from Western countries. The current study was undertaken to assess the effectiveness of these formulae in Japanese children. METHODS: We reviewed data for the past 5 years from children (0-8 years of age) intubated orally with the regular uncuffed ETT or RAE type. The data included the child's age in years and months and the internal diameter (ID) of the ETT. RESULTS: Data from 1301 children were reviewed. The IDs of the most frequently used ETT were 3.0 mm for patients less than 1 month old, 3.5 mm for those 1 to 4 months old, 4.0 mm for those 5 to 17 months old, and 4.5 mm for those 18 to 23 months old. In children 2 to 8 years old, the ID of the most frequently used ETT was in accordance with the ID predicted by the formula ID (mm) = [age in years + 16]/4. However, the ID predicted by this formula applied to only 53.5% of our patients. CONCLUSION: The age-based formula, ID (mm) = [age in years + 16]/4, is applicable to Japanese children. However, we recommend that three sizes be available before endotracheal intubation. PMID- 14566513 TI - Effects of ketamine on voltage-dependent calcium currents and membrane potentials in single bullfrog atrial cells. AB - PURPOSE: This study was undertaken to assess the effect of ketamine on L-type calcium channel current (I(Ca)) and membrane action potential in the bullfrog single atrial myocyte. METHODS: Bullfrog single atrial myocytes were prepared by enzymatic dispersion. Whole-cell voltage-clamp technique and current clamp technique were used to monitor I(Ca), membrane resting potential, and action potential. RESULTS: Ketamine (10(-5)-10(-3) M) showed dose-dependent inhibition of I(Ca) in a reversible manner. The 50% inhibitory concentration (IC(50)) of ketamine on I(Ca) was estimated to be 0.92 x 10(-5) M. Use-dependent block of I(Ca) was not observed. The resting membrane potential was depolarized at a high concentration (10(-4) M) of ketamine. Reduction of the plateau phase and prolonged duration of the action potential were observed in the presence of a high concentration of ketamine (10(-4) M). CONCLUSION: Ketamine has an inhibitory effect on I(Ca) in the bullfrog single atrial myocyte, and a high dose (10(-4) M) of ketamine prolonges the duration of the action potential. The mechanism of inhibition of I(Ca) seems to be a direct effect on the L-type calcium channel, not like an open channel blocker. PMID- 14566514 TI - Antinociceptive efficacy of antidepressants: assessment of five antidepressants and four monoamine receptors in rats. AB - PURPOSE: For assessment of the antinociceptive potency of antidepressants, we compared the antinociceptive effects of serotonin selective reuptake inhibitors (SSRIs) and classical tricyclic antidepressants (TCAs) in rats. We also attempted to elucidate the monoamine receptor subtypes predominantly involved in the antinociceptive effect of antidepressants. METHODS: Male Wistar rats received SSRIs (sertraline, fluvoxamine, and citalopram) or TCAs (imipramine and desipramine) intraperitoneally, and the reaction time until pain response in the hot plate test and licking time in the formalin test were measured 60 min later. We also observed the effects of prazosin (an alpha(1) antagonist), WB-4101 (a selective alpha(1A) antagonist), yohimbine (an alpha(2) antagonist), WAY-100635 (a selective 5-HT(1A) antagonist), and ketanserin (a 5-HT(2) antagonist), which were simultaneously administered with imipramine or desipramine, on the antidepressant-induced antinociceptive effect in the formalin test. RESULTS: In the hot plate test, desipramine, 20 mg.kg(-1), but not imipramine or sertraline, produced a significant increase in reaction time. In the formalin test, desipramine and imipramine produced significant reductions in the licking time at over 5 mg.kg(-1) and at over 10 mg.kg(-1), respectively. These reductions were nearly complete at 20 mg.kg(-1). On the other hand, both SSRIs induced significant reductions in the licking time only at 20 mg.kg(-1). Prazosin, WB 4101, and ketanserin significantly antagonized the antinociceptive effect of 10 mg.kg(-1) of imipramine. However, imipramine-induced antinociception was not affected by yohimbine and WAY-100635. Prazosin and ketanserin also significantly suppressed antinociception by 5 mg.kg(-1) of desipramine. CONCLUSION: These findings suggest that classical TCAs are likely to have a therapeutic advantage over SSRIs for pain control. In addition, it is likely that central alpha(1) adrenoceptors and 5-HT(2) receptors are predominantly involved in imipramine- and desipramine-induced antinociception. PMID- 14566515 TI - Effects of propofol and thiopental on the central nervous system during nociceptive stimulation in cats. AB - PURPOSE: Nociceptive stimulation may increase central nervous system (CNS) activity during anesthesia. However, it is not known whether propofol and thiopental have a similar inhibitory effect on the CNS during nociceptive stimulation. Therefore, we compared the antinociceptive effects of propofol and thiopental in cats. METHODS: In 12 cats, anesthesia was induced with 4% halothane in oxygen and maintained with 0.5% halothane in oxygen. The cortical electroencephalogram (EEG) and the electrical activity from the midbrain reticular neurons (R-MUA) were measured before and after sciatic nerve stimulation. The cats were then allocated to receive cumulative doses of either propofol (n = 6) or thiopental (n = 6) i.v. at 5-min intervals. Two minutes after each dose, the cortical EEG and the R-MUA were compared before and after sciatic nerve stimulation. RESULTS: Propofol and thiopental depressed the basal R-MUA to a similar degree at each dose. Sciatic nerve stimulation increased the R-MUA, and there were no differences in the maximum R-MUA values between propofol and thiopental. The cortical EEGs after each dose of anesthetic without stimulation showed similar patterns, and the patterns of change with stimulation were also similar for these two anesthetics. CONCLUSION: Propofol and thiopental have similar antinociceptive effects in cats. PMID- 14566516 TI - A case of tetralogy of Fallot with no neurological deficit after prolonged hypoxia. PMID- 14566517 TI - Pneumothorax of undetermined etiology occurring during general anesthesia. PMID- 14566518 TI - Spinal cord infarction in a patient undergoing left pyeloplasty in the right lateral kidney position. PMID- 14566519 TI - Successful relief of hip joint pain by percutaneous radiofrequency nerve thermocoagulation in a patient with contraindications for hip arthroplasty. PMID- 14566520 TI - Pyogenic spondylitis and epidural abscess distant from the site of continuous epidural block in a patient with postherpetic neuralgia. PMID- 14566521 TI - Clinical application of amantadine, an NMDA antagonist, for neuropathic pain. PMID- 14566522 TI - Severe sinoatrial dysfunction after esophageal surgery: a case report. PMID- 14566524 TI - Femoral arterial hypotension secondary to the placement of a pediatric intraaortic balloon. PMID- 14566523 TI - Epidural injection with saline for treatment of postspinal headache: comparison with epidural blood patch. PMID- 14566525 TI - Effects of epidural fentanyl and intravenous flurbiprofen for visceral pain during cesarean section under spinal anesthesia. AB - PURPOSE: Despite adequate levels of sensory blockade, patients sometimes complain of abdominal pain during cesarean section performed under spinal anesthesia. The aim of this study was to evaluate the effects of epidural fentanyl and intravenous flurbiprofen on visceral pain during cesarean section in patients having spinal anesthesia. METHODS: Thirty ASA physical status I and II patients undergoing elective cesarean section were studied. Spinal-epidural anesthesia was performed in all groups. Group A received no additional analgesics, group B received epidural fentanyl 100 mug, and group C received flurbiprofen 50 mg i.v. immediately after the delivery. Postdelivery, intraoperative visceral pain was evaluated by using the visual analog scale. Incidence and visual analog scale scores of visceral pain and incidence of intraoperative nausea and vomiting were obtained from each patient. RESULTS: Visual analog scale scores of pain were significantly lower in group B than in the other groups (P < 0.05). The incidence of nausea was comparable in all groups. The incidence of intraoperative vomiting was lower in group C than in the other groups (P < 0.05). CONCLUSION: Epidural fentanyl, but not intravenous flurbiprofen, decreases the incidence and severity of visceral pain during cesarean section. PMID- 14566526 TI - QT intervals and QT dispersion in patients with subarachnoid hemorrhage. AB - PURPOSE: To clarify the clinical significance of QT dispersion and the longest QT intervals (L-QTc) in patients with subarachnoid hemorrhage (SAH). METHODS: ECGs, clinical features, and laboratory data were analyzed in 38 patients with SAH (R) and 30 with unruptured cerebral aneurysms (U). Standard 12-lead ECGs obtained on admission were analyzed manually, and the longest QT interval (L-QTc) and the QT dispersion (difference between longest and shortest QTc) were compared between groups. RESULTS: There were no differences between groups R and U in age, sex, or location of aneurysms. The QT dispersion and L-QTc were greater in R than in U (109 +/- 49 vs 64 +/- 21 ms and 503 +/- 63 vs 435 +/- 38 ms, respectively; P < 0.01). The QT dispersion and L-QTc were longer in patients with premature ventricular contractions (PVCs) than in patients without PVCs (185 +/- 30 vs 85 +/- 41 ms and 586 +/- 47 vs 467 +/- 59 ms, respectively; P < 0.01). There were positive correlations between QT dispersion or L-QTc and preoperative Hunt and Hess grade (rs = 0.560 and rs = 0.615, respectively; P < 0.01). QT dispersion and L-QTc tended to correlate negatively with serum K(+) (r = -0.365 and r = -0.376, respectively). CONCLUSION: QT dispersion in patients with SAH is prolonged, especially in high-grade cases. PMID- 14566527 TI - Clonidine premedication prevents sympathetic hyperactivity but does not prevent hypothalamo-pituitary-adrenocortical responses in patients undergoing laparoscopic cholecystectomy. AB - PURPOSE: The effects of oral administration of clonidine on sympathetic and endocrinological responses were investigated in patients undergoing elective laparoscopic cholecystectomy. METHODS: Twenty adult patients were allocated randomly to the clonidine group (n = 10) or the control group (n = 10). The control and clonidine groups received placebo on clonidine 4 microg.kg(-1) orally 2 h before the induction of anesthesia. All patients underwent laparoscopic cholecystectomy under isoflurane anesthesia. The hemodynamic variables were observed perioperatively. Plasma concentrations of cortisol, ACTH, noradrenaline, adrenaline, and dopamine were determined before administration of clonidine or placebo, 2 h after the beginning of the operation, and 3 h after the end of the operation. RESULTS: Systolic and diastolic blood pressures were lower in the clonidine group than in the control group immediately after endotracheal intubation and extubation (P < 0.05). Patients in the clonidine group showed lower plasma concentrations of noradrenaline 2 h after the beginning of the operation than patients in the control group (P < 0.01). However, the plasma concentrations of the other hormones did not differ between groups. CONCLUSION: Clonidine premedication prevents sympathetic hyperactivity but does not suppress hypothalamo-pituitary-adrenocortical responses in patients undergoing laparoscopic cholecystectomy. PMID- 14566528 TI - Power spectral analysis of the electroencephalogram during induced total spinal block. AB - PURPOSE: To evaluate the effects of total spinal block (TSB) on brain function, TSB-induced changes in cortical electrical activities were analyzed using power spectral analysis of an electroencephalogram (EEG). METHODS: Six patients suffering from chronic pain who were undergoing TSB therapy were studied. TSB was established with intrathecal 1% lidocaine (0.3 ml.kg(-1)) injected through the C1 2 lateral intervertebral space. Mechanical ventilation was continued via a laryngeal mask until the recovery of respiration. The EEG recording was started before TSB induction and continued until 10 min after extubation. The following processed EEG parameters were monitored: spectral edge frequency-90% (SEF90), spectral median frequency (SMF), and relative power in the frequency bands of delta, theta, alpha, beta, and the delta ratio [(alpha + beta)/delta]. RESULTS: TSB induced an unconscious state more than 40 min in all patients. During TSB, 12 13 Hz in SEF90 and 6-7 Hz in SMF were observed. These values are consistent with the previously reported prearousal threshold from general anesthesia. The other EEG descriptors did not change during the TSB-induced unconscious state. CONCLUSION: The dissociation of cortical electrical activities and the clinical coma-like condition may be characteristic of the TSB-induced unconscious state. PMID- 14566529 TI - The relationship of brain catecholamine levels to enflurane requirements among three strains of mice with different anesthetic sensitivities. AB - PURPOSE: . It has been reported that brain catecholamines alter the minimum alveolar concentration (MAC) of anesthetics. The extent of the relation between the levels of brain catecholamine and anesthetic sensitivity should be evaluated by excluding several factors. METHODS: Anesthetic sensitivity was measured by using loss of the righting reflex in three strains of mice with different sensitivities. The mice were decapitated without any anesthesia, adding on ddN and C57BL/6J mice in 2% enflurane, their brains were divided into three parts, and dopamine and norepinephrine levels were analyzed by high-performance liquid chromatography (HPLC). Results. The values of enflurane requirement (%) were 1.30 +/- 0.05 in ddN, 1.10 +/- 0.02 in C57BL/6J, and 1.05 +/- 0.02 in MSM mice. The values of dopamine (microg.g(-1)) in the mesencephalon were 0.23 +/- 0.02 in ddN, 0.15 +/- 0.02 in C57BL/6J, and 0.12 +/- 0.02 in MSM (mean +/- SE). No statistical significance in the values in 2% enflurane could be obtained between ddN and C57BL/6J. The stepwise regression line showed a significant correlation: enflurane requirement (%) = -0.89 + 1.60 x (dopamine levels of mesencephalon) (r(2) = 0.571, P < 0.0001). CONCLUSION: Dopamine in the mesencephalon seems to play an important role in the production of different anesthetic sensitivities, and the anesthetic mechanism might be related to the regulation of dopamine levels that promote arousal. PMID- 14566530 TI - Clinical characteristics of perioperative coronary spasm: reviews of 115 case reports in Japan. AB - PURPOSE: Factors affecting perioperative development of coronary spasm have not been elucidated. A number of case reports describing perioperative coronary spasm have appeared in Japanese anesthesia journals, mostly published in Japanese. The purpose of this study was to investigate the contributing factors affecting perioperative coronary artery spasm by reviewing the published articles. METHODS: Reports were identified by using Medline database (1968-1998) or by manually searching nonindexed Japanese journals. The clinical characteristics of perioperative coronary spasm were analyzed in 115 patients who developed coronary artery spasm during the perioperative period. RESULTS: The mean age of the patients was 64 +/- 9 years (range, 36 to 87 years). There were 97 men (84%) and 18 women (16%). Preoperative risk factors included hypertension (27%), angina pectoris (27%), cigarette smoking (13%), and diabetes mellitus (11%). The attack was related to inadequate depth of general anesthesia (23%), use of vasopressors (22%), vagal reflex (19%), administration of drugs other than vasopressors (17%), and epidural block (15%). About 85% of patients showed no ischemic abnormality on the preoperative electrocardiogram, whereas 56% had significant coronary stenosis on postoperative coronary arteriography. Coronary spasm tended to occur in patients under inhalation anesthesia combined with epidural block. Nitrates alleviated the episode in the majority of cases, whereas defibrillation and cardiac massage were required in 19% of patients. No deaths were reported. CONCLUSION: Perioperative coronary spasm is prevalent in elderly male patients with coronary risk factors who undergo abdominal or thoracic surgery under inhalational anesthesia combined with epidural anesthesia. Instability of the autonomic nervous system and/or vascular hyperreactivity may be the underlying pathogenic mechanisms of perioperative coronary spasm. PMID- 14566531 TI - Use of a processed endotracheal tube in general anesthesia for palatoplasty in a patient with subglottic stenosis. PMID- 14566532 TI - Growth causes difficult tracheal intubation in a patient with Kniest dysplasia. PMID- 14566533 TI - A case of cauda equina syndrome following spinal anesthesia with hyperbaric dibucaine. PMID- 14566534 TI - Anesthetic management of a patient with McCune-Albright syndrome accompanied by polyostotic fibrous osteodysplasia. PMID- 14566535 TI - Two cases of chemical meningitis following spinal anesthesia. PMID- 14566536 TI - The Ombredanne inhaler in Japan. PMID- 14566538 TI - The incidence of "coring" during aspiration of propofol from a 50-ml vial. PMID- 14566537 TI - Usefulness of midazolam premedication for volatile induction of anesthesia in adults. PMID- 14566539 TI - Self-reported recovery time of daily activity after bone marrow harvesting from healthy donors. AB - PURPOSE: The study was planned to describe and compare the resumption of daily activity after bone marrow harvesting (BMH) under general anesthesia with different anesthetic agents, isoflurane, sevoflurane and propofol. METHODS: Sixty five adult healthy allogenic donors (26 women, 39 men; 37.0 +/- 9.5 years of age; ASA class I or II) were studied. BMH was performed under general anesthesia with isoflurane, sevoflurane, or propofol as the main anesthetic agent. On day 1, donors were asked how soon they could resume five daily activities: talking, walking, drinking, eating, and reading. RESULTS: Although there was considerable variation in resumption time within and among these five activities, 77% of donors resumed all of these basic daily activities within 12 h after BMH. Donors who received propofol resumed talking significantly faster than those who received sevoflurane or isoflurane, but the choice of main anesthetic agent did not affect the time of resumption of other activities. Lower preoperative hemoglobin concentration and greater decline of hemoglobin concentration on day 1 were significantly associated with slower resumption of walking and reading. CONCLUSION: Although the choice of main anesthetic agent and anemia affected postoperative recovery, 77% of donors could resume these five daily activities within 12 h. PMID- 14566540 TI - The effect of midazolam-thiopental coinduction on recovery in minor surgery. AB - PURPOSE: The effects of midazolam-thiopental coinduction on recovery were investigated and compared with thiopental induction. METHODS: Fifty patients, ASA 1 or 2, undergoing minor orthopedic surgery, were randomly divided into coinduction and thiopental groups. During preoxygenation, the patients received midazolam 0.1 mg.kg(-1) (coinduction group) or saline (thiopental group) 1 min before induction of anesthesia with thiopental. Isoflurane and nitrous oxide were used to maintain anesthesia. Isoflurane concentration was adjusted to keep blood pressure within +/-20% of the preoperative value. The time to awaken (open eyes, give name and birth-date) and the time to discharge readiness were recorded. Psychomotor tests, including simple light reaction time (SLRT), sedation analogue scale (SAS), and digit span test, were performed pre- and postoperatively. RESULTS: The induction dose of thiopental was significantly lower in the coinduction group. End-tidal isoflurane concentration during surgery was also lower in the coinduction group. There were no significant differences in awakening times and discharge readiness between the two groups. Although SAS values were lower in the coinduction group than in the thiopental group 8 and 24 h after anesthesia, other test results were similar in both groups. The frequency of nausea and vomiting in the recovery period was lower in the coinduction group. CONCLUSION: We conclude that midazolam-thiopental coinduction is a suitable technique when used in conjunction with isoflurane in day-case surgery. PMID- 14566541 TI - Age-corrected intraoperative tachycardia correlates with postoperative electrocardiographic alterations. AB - PURPOSE: The intrinsic heart rate (IHR) has been calculated by an age-dependent formula; 118.1 - 0.57 x Age (years). The aim of this study was to examine whether intraoperative tachycardia of different criteria correlates with the incidence of alterations of postoperative electrocardiograms (ECG). METHODS: One hundred and twenty-two patients who underwent elective laparotomy in general surgery were studied. Tachycardia was defined as a heart rate of either 100, 110, 120, or 130 beats per minute or more, and heart rates of 1.1, 1.2, 1.3, or 1.4 times or more of the IHR. Postoperative ECG alterations were defined as positive when (1) one or more of the first letters of Minnesota Code I, IV, and V were newly added, or (2) Q-waves extended to an adjacent lead, or (3) new ST-segment depression or elevation of 0.1 mV or more was observed at 80 ms beyond the ST-junction. Correlations between intraoperative tachycardia of different criteria and alterations in 12-h postoperative ECG were investigated. RESULTS: The incidence of postoperative ECG alterations was significantly greater when the intraoperative heart rate exceeded 1.2 IHR (143 - 0.7 x Age) for 30 min and 1.3 IHR (151 - 0.7 x Age) for 5 min (P = 0.04 and P = 0.003, respectively). CONCLUSION: Age-corrected intraoperative tachycardia showed a good correlation with the incidence of postoperative ECG alterations. PMID- 14566542 TI - Celite-activated viscometer Sonoclot can measure the suppressive effect of tranexamic acid on hyperfibrinolysis in cardiac surgery. AB - PURPOSE: To investigate the usefulness of the celite-activated viscometer Sonoclot for monitoring fibrinolytic status in cardiac surgery, we demonstrated the effectiveness of high doses of tranexamic acid, an antifibrinolytic agent, in reducing postoperative bleeding. METHODS: Thirty-two American Society of Anesthesiologists (ASA) physical status III patients who required cardiac surgery with cardiopulmonary bypass (CPB) were studied. Anesthesia was induced by a high dose of fentanyl and midazolam with oxygen and was maintained by the intermittent administration of these agents. Patients were divided into two groups: the control group (n = 15) and patients receiving tranexamic acid (TA; n = 17). The TA group received a high dose (50 mg/kg) of TA twice, once before and once after CPB. The percentage diminishing rate of the Sonoclot tracing 15 min after maximum clot signal (DR(15)) and the amount of postoperative bleeding were measured. RESULTS: After CPB, DR(15) in the control group (mean 28.3%) increased significantly by 45%, and the DR(15) in the TA group (16.1%) was significantly lower than that in the control group. The amount of postoperative bleeding in the TA group (546 ml) was significantly less, by 34%, than that in the control group (829 ml). CONCLUSION: Prophylactic administration of high-dose TA in cardiac surgery reduces postoperative bleeding, and this effect is consistent with changes in the diminishing rate using Sonoclot. The celite-activated viscometer Sonoclot is recommended for use in cardiac surgery for rapid assessment of fibrinolytic status. PMID- 14566543 TI - Sevoflurane reduces dysrhythmias during reperfusion in the working rat heart. AB - PURPOSE: The effects of sevoflurane on myocardial reperfusion injury have not been well studied. The purpose of this study was to determine the effects of sevoflurane on myocardial function, arrhythmia, and metabolism during reperfusion in an isolated working rat heart model. METHODS: Thirty-two hearts were divided into four groups according to the timing of 2.5% sevoflurane administration: group I, control, no sevoflurane; group II, sevoflurane administered only before ischemia; group III, sevoflurane only during reperfusion; group IV, sevoflurane during the whole study period. Myocardial contractility, myocardial ATP, lactate, and glycogen levels were assessed in the reperfusion period following global heart ischemia of 15 min duration. The incidence and duration of ventricular fibrillation were also observed in the reperfusion period. RESULTS: There was no difference in cardiac output and left ventricular dP/ dt max among the four groups at 10, 15, and 20 min after reperfusion. There was no difference in myocardial ATP, lactate and glycogen contents between the groups. The incidences of ventricular fibrillation during reperfusion were 100%, 63%, 100%, and 25% (P < 0.05 vs control), and the durations of ventricular fibrillation during reperfusion were 375 +/- 269, 104 +/- 98 (P < 0.05 vs control), 303 +/- 189, and 93 +/- 245 (P < 0.05 vs control) in groups I, II, III, and IV, respectively (mean +/- SD). CONCLUSION: The administration of sevoflurane prior to reperfusion appears to provide myocardial protection, as assessed by reduced dysrhythmias during reperfusion. PMID- 14566544 TI - Hyperbaric hyperoxia suppresses growth of Staphylococcus aureus, including methicillin-resistant strains. AB - PURPOSE: We investigated the effects of increased oxygen tension on the in vitro growth of Staphylococcus aureus (MRSA), methicillin-sensitive Staphylococcus aureus (MSSA), and Escherichia coli (E. coli). METHODS: The effects of oxygen tension [normobaric normoxia (21% O(2) at 1 atm), normobaric hyperoxia (100% O(2) at 1 atm), hyperbaric normoxia (21% O(2) at 2 atm), and hyperbaric hyperoxia (100% O(2) at 2 atm) on the in vitro growth of MRSA, MSSA, and E. coli were investigated by population analysis. RESULTS: Compared with normobaric normoxia, a 90-min exposure to hyperbaric hyperoxia significantly inhibited growth of both MRSA (by 25.0 +/- 3.0%, mean +/- SEM; P < 0.01) and MSSA (by 24.0 +/- 3.3%; P < 0.01). Normobaric hyperoxia and hyperbaric normoxia were without effect. In contrast, the growth of E. coli was not affected by any of the above treatments. CONCLUSION: Our results indicate that the bacterium S. aureus, including resistant strains, is susceptible to oxygen stress. The observation that relatively brief (90-min) treatment with hyperbaric hyperoxia is sufficient to produce significant growth inhibition suggests that hyperbaric hyperoxia may be useful in the treatment of serious staphylococcal infections. PMID- 14566545 TI - Effects of ulinastatin (urinary trypsin inhibitor) on ATP, intracellular pH, and intracellular sodium transients during ischemia and reperfusion in the rat kidney in vivo. AB - PURPOSE: To investigate the effects of ulinastatin on renal ischemia-reperfusion injury, we monitored the dynamic changes in ATP, intracellular pH (pHi), and intracellular sodium (Nai) in rats in vivo. METHODS: Renal ischemia was induced by clamping the abdominal aorta for 30 min followed by reperfusion for 60 min. Ulinastatin, 50,000 U.kg(-1) (UTI group), or normal saline (NS group) was infused for 30 min before ischemia. (31)P- and double quantum (23)Na-NMR were used to monitor ATP, pHi, and Nai. RESULTS: During ischemia, ATP was rapidly depleted and Nai increased to the same extent in both groups. After 60 min reperfusion, Nai in the NS group was almost restored to the preischemic baseline level (117.2 +/- 7.4% of the baseline value), but the recovery of ATP was incomplete (60.9 +/- 7.7%). The recovery of Nai in the UTI group began earlier than in the NS group with better recovery of ATP. The pHi values showed severe acidosis in the NS group compared with the UTI group during ischemia and reperfusion. As for ultrastructural findings, after 60 min reperfusion, the mitochondria were less swollen and less disorganized with respect to the membrane and the cristae in the UTI group. CONCLUSION: The transcellular sodium gradient is restored before the ATP level is normalized during postischemic reperfusion. Ulinastatin might protect mitochondrial conformation during ischemia, and facilitate functional recovery of the ionic pump after reperfusion. PMID- 14566547 TI - Transient renal tubular dysfunction in a patient with severe asthmatic attack treated with sevoflurane. PMID- 14566546 TI - A possible role for glutamate receptor-mediated excitotoxicity in chronic pain. PMID- 14566548 TI - Anesthetic management of a patient with Klippel-Feil syndrome. PMID- 14566549 TI - One-lung ventilation with the Univent tube in a pediatric patient undergoing video-assisted thoracoscopic surgery. PMID- 14566550 TI - Laryngeal mask airway is useful for fiberoptic bronchoscopic evaluation of subglottic stenosis in children: a report of eleven cases. PMID- 14566551 TI - Combined propofol and thoracic epidural anesthesia for pulmonary resection in a morbidly obese patient with sleep apnea syndrome. PMID- 14566552 TI - Anesthetic management of the patient with systemic lupus erythematosus and severe granulocytopenia. PMID- 14566553 TI - Performance prediction of industrial centrifuges using scale-down models. AB - Computational fluid dynamics was used to model the high flow forces found in the feed zone of a multichamber-bowl centrifuge and reproduce these in a small, high speed rotating disc device. Linking the device to scale-down centrifugation, permitted good estimation of the performance of various continuous-flow centrifuges (disc stack, multichamber bowl, CARR Powerfuge) for shear-sensitive protein precipitates. Critically, the ultra scale-down centrifugation process proved to be a much more accurate predictor of production multichamber-bowl performance than was the pilot centrifuge. PMID- 14566554 TI - Analysis of the behaviour of Dictyostelium discoideum in immobilised state by means of continuous cultivation. AB - The social amoeba Dictyostelium discoideum is amenable to cultivation in the immobilised state most simply by colonisation of porous supports. An analysis of the growth behaviour of D. discoideum in the immobilised state is reported. For this purpose, D. discoideum was cultivated in continuously operated reactors in a suspension culture (homogeneous system) and immobilised on a porous support (heterogeneous system). Thus, it is possible to compare homogeneous and heterogeneous systems under steady-state conditions. Immobilisation was achieved by the colonisation of porous glass beads (SIRAN). Simple models are applied in order to describe the growth behaviour of fractions of both the cells in free suspension and the cells inside the porous carrier. This analysis shows that D. discoideum inside the pores grows at a rate of only about 10% compared with that in free solution. The consequence of this behaviour is discussed in terms of reactor performance. PMID- 14566555 TI - Impact of herbivores on nitrogen cycling: contrasting effects of small and large species. AB - Herbivores are reported to slow down as well as enhance nutrient cycling in grasslands. These conflicting results may be explained by differences in herbivore type. In this study we focus on herbivore body size as a factor that causes differences in herbivore effects on N cycling. We used an exclosure set-up in a floodplain grassland grazed by cattle, rabbits and common voles, where we subsequently excluded cattle and rabbits. Exclusion of cattle lead to an increase in vole numbers and a 1.5-fold increase in net annual N mineralization at similar herbivore densities (corrected to metabolic weight). Timing and height of the mineralization peak in spring was the same in all treatments, but mineralization in the vole-grazed treatment showed a peak in autumn, when mineralization had already declined under cattle grazing. This mineralization peak in autumn coincides with a peak in vole density and high levels of N input through vole faeces at a fine-scale distribution, whereas under cattle grazing only a few patches receive all N and most experience net nutrient removal. The other parameters that we measured, which include potential N mineralization rates measured under standardized laboratory conditions and soil parameters, plant biomass and plant nutrient content measured in the field, were the same for all three grazing treatments and could therefore not cause the observed difference. When cows were excluded, more litter accumulated in the vegetation. The formation of this litter layer may have added to the higher mineralization rates under vole grazing, through enhanced nutrient return through litter or through modification of microclimate. We conclude that different-sized herbivores have different effects on N cycling within the same habitat. Exclusion of large herbivores resulted in increased N annual mineralization under small herbivore grazing. PMID- 14566556 TI - Female survival, lifetime reproductive success and mating status in a passerine bird. AB - In facultatively polygynous birds, secondary females of polygynously mated males typically have reduced annual reproductive success, because polygynous males provide less paternal care than monogamous males. Life history theory predicts that, as a result of increased reproductive investment, secondary females should suffer from reduced survival and lifetime reproductive success, but previous studies provided only weak support for this hypothesis. We used 7 years of data to study the fitness of female collared flycatchers Ficedula albicollis in relation to mating status by estimating survival and lifetime reproductive success. Taking differences in recapture probability into account, a mark recapture analysis revealed that females observed at least once to breed as secondary female had higher survival than other females. This relationship was not confounded by laying date, because when we assessed the impact of laying date on survival, we found similar survival patterns. Females of polygynous males had reduced breeding success in terms of number of young fledged during the current reproductive event. However, during their lifetime females found at least once in primary or secondary mating status produced significantly more eggs, and at least the same number of fledglings and recruits as monogamous females. Thus, in the collared flycatcher, females of polygynously mated males seem to suffer from mating status during the most recent reproductive event, but considering survival and lifetime reproductive success, the apparently disadvantageous mating event is not necessarily associated with reduced residual reproductive value. PMID- 14566557 TI - Colony-structure variation and interspecific competitive ability in the invasive Argentine ant. AB - The success of some invasive species may depend on phenotypic changes that occur following introduction. In Argentine ants (Linepithema humile) introduced populations typically lack intraspecific aggression, but native populations display such behavior commonly. We employ three approaches to examine how this behavioral shift might influence interspecific competitive ability. In a laboratory experiment, we reared colonies of Forelius mccooki with pairs of Argentine ant colonies that either did or did not exhibit intraspecific aggression. F. mccooki reared with intraspecifically non-aggressive pairs of Argentine ants produced fewer eggs, foraged less actively, and supported fewer living workers than those reared with intraspecifically aggressive pairs. At natural contact zones between competing colonies of L. humile and F. mccooki, the introduction of experimental Argentine ant colonies that fought with conspecific field colonies caused L. humile to abandon baits in the presence of F. mccooki, whereas the introduction of colonies that did not fight with field colonies of Argentine ants resulted in L. humile retaining possession of baits. Additional evidence for the potential importance of colony- structure variation comes from the Argentine ant's native range. At a site along the Rio de la Plata in Argentina, we found an inverse relationship between ant richness and density of L. humile (apparently a function of local differences in colony structure) in two different years of sampling. PMID- 14566558 TI - Secondary succession is influenced by belowground insect herbivory on a productive site. AB - We investigated the effects of insect herbivory on a plant community of a productive old-field community by applying foliar and soil insecticides in a full factorial design. During the first 3 years of succession, insecticide treatments had only minor effects on total cover abundance and species richness. However, species ranking within the plant community was strongly affected by soil insecticide but not by foliar insecticide. Creeping thistle, Cirsium arvense, dominated the experimental plots with reduced root herbivory, while square stemmed willow-herb, Epilobium adnatum, dominated the control and the plots with foliar insecticide. When soil insecticide was applied, cover abundance of monocarpic forbs increased and cover abundance of polycarpic herbs decreased compared to the control. However, this effect was due to a few abundant plant species and is not based on a consistent difference between life history groups. Instead, application of soil insecticide promoted persistence of species that established at the start of succession, and suppressed species that established in the following years. We conclude that below-ground herbivory reduces competitive ability of resident species and, thus, facilitates colonization by late-successional species. Hence, soil insects can exert strong top-down effects on the vegetation of productive sites by affecting dominant plant species and altering competitive balances. PMID- 14566559 TI - Molecular analysis of congenital central hypoventilation syndrome. AB - Congenital central hypoventilation syndrome (CCHS or Ondine's curse; OMIM 209880) is a disorder characterized by an idiopathic failure of the automatic control of breathing. CCHS is frequently complicated with neurocristopathies such as Hirschsprung's disease (HSCR). The genes involved in the RET-GDNF signaling and/or EDN3-EDNRB signaling pathways have been analyzed as candidates for CCHS; however, only a few patients have mutations of the RET, EDN3, and GDNF genes. Recently, mutations of the PHOX2B gene, especially polyalanine expansions, have been detected in two thirds of patients. We studied the RET, GDNF, GFRA1, PHOX2A, PHOX2B, HASH-1, EDN1, EDN3, EDNRB, and BDNF genes in seven patients with isolated CCHS and three patients with HSCR. We detected polyalanine expansions and a novel frameshift mutation of the PHOX2B gene in four patients and one patient, respectively. We also found several mutations of the RET, GFRA1, PHOX2A, and HASH 1 genes in patients with or without mutations of the PHOX2B gene. Our study confirmed the prominent role of mutations in the PHOX2B gene in the pathogenesis of CCHS. Mutations of the RET, GFRA1, PHOX2A, and HASH-1 genes may also be involved in the pathogenesis of CCHS. To make clear the pathogenesis of CCHS, the analysis of more cases and further candidates concerned with the development of the autonomic nervous system is required. PMID- 14566560 TI - Protein N-glycosylation is similar in the moss Physcomitrella patens and in higher plants. AB - We have investigated the structure of glycans N-linked to the proteins of the moss Physcomitrella patens. The structural elucidation was carried out by western blotting using antibodies specific for N-glycan epitopes and by analysis of N linked glycans enzymatically released from a total protein extract by combination of MALDI-TOF and MALDI-PSD mass spectrometry analysis. Nineteen N-linked oligosaccharides were characterised ranging from high-mannose-type and truncated paucimannosidic-type to complex-type N-glycans harbouring core-xylose, core alpha(1,3)-fucose and Lewis(a), as previously described for proteins from higher plants. This demonstrates that the processing of N-linked glycans, as well as the specificity of glycosidases and glycosyltransferases involved in this processing, are highly conserved between P. patens and higher plants. As a consequence, P. patens appears to be a new promising model organism for the investigation of the biological significance of protein N-glycosylation in the plant kingdom, taking advantage of the potential for gene targeting in this moss. PMID- 14566561 TI - Maltose is the major form of carbon exported from the chloroplast at night. AB - Transitory starch is formed in chloroplasts during the day and broken down at night. We investigated carbon export from chloroplasts resulting from transitory starch breakdown. Starch-filled chloroplasts from spinach ( Spinacia oleracea L. cv. Nordic IV) were isolated 1 h after the beginning of the dark period and incubated for 2.5 h, followed by centrifugation through silicone oil. Exported products were measured in the incubation medium to avoid measuring compounds retained inside the chloroplasts. Maltose and glucose made up 85% of the total exported products and were exported at rates of 626 and 309 nmol C mg(-1) chlorophyll h(-1), respectively. Net export of phosphorylated products was less than 5% and higher maltodextrins were not detected. Maltose levels in leaves of bean ( Phaseolus vulgaris L. cv. Linden), spinach, and Arabidopsis thaliana (L.) Heynh. were low in the light and high in the dark. Maltose levels remained low and unchanged during the light/dark cycle in two starch-deficient Arabidopsis mutants, stf1, deficient in plastid phosphoglucomutase, and pgi, deficient in plastid phosphoglucoisomerase. Through the use of nonaqueous fractionation, we determined that maltose was distributed equally between the chloroplast and cytosolic fractions during darkness. In the light there was approximately 24% more maltose in the cytosol than the chloroplast. Taken together these data indicate that maltose is the major form of carbon exported from the chloroplast at night as a result of starch breakdown. We hypothesize that the hydrolytic pathway for transitory-starch degradation is the primary pathway used when starch is being converted to sucrose and that the phosphorolytic pathway provides carbon for other purposes. PMID- 14566562 TI - Length of time in tissue culture can affect the selected glyphosate resistance mechanism. AB - Usually, stepwise selection of plant suspension cultures with gradually increasing concentrations of the herbicide glyphosate results in the amplification of the target enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS; EC 2.5.1.19) gene that leads to resistance by increasing EPSPS mRNA and enzyme activity. We show that glyphosate selection with newly initiated suspension cultures can produce resistant lines with resistance mechanisms other than gene amplification and that usually as the cultures age gene amplification becomes the predominant mechanism. Gene amplification did not occur in 3 lines selected from 5-month-old Datura innoxia Mill. cultures but did occur in all 10 lines selected after 52 months. Selection with Nicotiana tabacum L. (tobacco) less than 5 months old produced 2 lines out of 24 with no EPSPS amplification while all 17 lines selected from older cultures contained amplified genes. Lines selected from the oldest culture (35 years) also exhibited amplification of several different genes, indicating the expression of different EPSPS genes or an enhanced gene amplification incidence. None of the 15 lines selected from 2 different 5-month-old Daucus carota L. (carrot) lines exhibited amplification while amplification led to the resistance of all 7 lines selected from one of the original carrot lines (DHL) after 3 years. However, the other line (Car4) was exceptional and produced only non-amplified lines (9 of 9) after 8 years in culture. These results show that plant tissue cultures change with time in culture and that several different new mechanisms can result in glyphosate resistance. PMID- 14566564 TI - Modulation of H reflexes in the forearm during voluntary teeth clenching in humans. AB - We investigated whether there is any modulation of the H reflex in the forearm during teeth clenching and how any correlation that may be found is modulated. The H reflexes of the flexor carpi radialis (FCR) and the extensor carpi radialis (ECR) muscles were evoked on the right side in five healthy adult volunteers. The H reflexes of the FCR and ECR muscles were facilitated in association with voluntary teeth clenching in a force-dependent manner ( r=0.46-0.663, P<0.05). The increase in amplitude of the H reflex of the FCR muscle associated with teeth clenching started before the onset of the EMG activity of the masseter muscle. The results of the present study demonstrate that oral motor activity exerts strong influences on the motor activity of the forearm. PMID- 14566565 TI - Relationship between double product break point, lactate threshold, and ventilatory threshold in cardiac patients. AB - A double product break point (DPBP) occurs simultaneously with both ventilatory threshold (VT) and lactate threshold (LT) in normal subjects. We sought to determine whether a DPBP also occurs in cardiac patients and to investigate correlations between DPBP, VT and LT (study 1). We also evaluated a non-invasive DPBP measurement system that determined blood pressure automatically by the cuff method (study 2). Study 1 comprised 15 patients [8 men and 7 women, mean (SD) age 47.7 (11.1) years] who performed cardiopulmonary exercise tests on a cycle ergometer. The double product was defined as the product of heart rate and direct systolic blood pressure. Arterial blood samples were obtained for measurement of lactate, pyruvate, pH, and norepinephrine levels. VT was determined by gas analysis, and LT was determined as the lactate/pyruvate ratio. DPBPs were detected in all 15 patients. Double product slopes above the DPBP were significantly greater than those below the DPBP (286.2 vs 98.5/W, P<0.001). The lactate/pyruvate ratio increased, arterial pH decreased, and plasma norepinephrine concentration increased above the DPBP. DPBP had strong positive correlations with VT ( r=0.93) and LT ( r=0.95). Study 2 comprised 65 cardiac patients. The DPBP was detected in 89.2% of patients and correlated closely with VT. We conclude that DPBP occurs near VT and LT in cardiac patients during incremental exercise, that the noninvasive DPBP measurement method is comparable to the invasive method, and that DPBP may be as useful an index of exercise intensity in patients with cardiac disease as VT or LT. PMID- 14566566 TI - Limitation of muscle deoxygenation in the triceps during incremental arm cranking in women. AB - The present study investigated the difference in oxygen kinetics in the exercising muscle between arm cranking and leg cycling in women. Twenty-seven females completed incremental arm cranking and leg cycling tests on separate days. During each exercise, spatially resolved near-infrared spectroscopy was used to measure changes in the tissue oxygen saturation ( SO(2)), oxygenated (oxy ) hemoglobin and/or myoglobin (Hb/Mb), deoxygenated (deoxy-) Hb/Mb, and total Hb/Mb in the triceps during arm cranking and in the vastus lateralis during leg cycling. During arm cranking, there was a rapid increase in the respiratory exchange ratio and a lower ventilatory threshold compared to leg cycling, which confirmed accelerated anaerobic glycolysis in this mode of exercise. During leg cycling, SO(2) remained decreased near to or until approaching peak oxygen uptake (VO(2peak)). During arm cranking, however, the decrease in oxy-Hb/Mb and increase in deoxy-Hb/Mb stopped at the middle of VO(2peak) (mean 51.4%), consequently resulting in a leveling off in the SO(2 )decrease, although total Hb/Mb continued to increase. These results might suggest that the oxygen demand in the triceps attained the maximum at that intensity, despite an adequate oxygen supply during arm cranking. PMID- 14566567 TI - Influence of muscle fibre type and pedal rate on the VO2-work rate slope during ramp exercise. AB - We hypothesised that the ratio between the increase in oxygen uptake and the increase in work rate (Delta VO(2)/DeltaWR) during ramp cycle exercise would be significantly related to the percentage type II muscle fibres at work rates above the gas exchange threshold (GET) where type II fibres are presumed to be active. We further hypothesised that ramp exercise at higher pedal rates, which would be expected to increase the proportional contribution of type II fibres to the total power delivered, would increase the Delta VO(2)/DeltaWR slope at work rates above the GET. Fourteen healthy subjects [four female; mean (SD): age 25 (3) years, body mass 74.3 (15.1) kg] performed a ramp exercise test to exhaustion (25 W min( 1)) at a pedal rate of 75 rev min(-1), and consented to a muscle biopsy of the vastus lateralis. Eleven of the subjects also performed two further ramp tests at pedal rates of 35 and 115 rev min(-1). The Delta VO(2)/DeltaWR slope for exercise GET ( S(2)) was significantly correlated to percentage type II fibres ( r=0.54; P=0.05). The ratio between the Delta VO(2)/DeltaWR slopes for exercise above and below the GET ( S(2)/ S(1)) was significantly greater at the pedal rate of 115 rev min(-1) [1.22 (0.09)] compared to pedal rates of 35 rev min(-1) [0.96 (0.02)] and 75 rev min(-1) [1.09 (0.05), ( P<0.05)]. The greater increase in S(2) relative to S(1) in subjects (1) with a high percentage type II fibres, and (2) at a high pedal rate, suggests that a greater recruitment of type II fibres contributes in some manner to the "xs" VO(2) observed during ramp exercise. PMID- 14566568 TI - Oxidation of [13C]glucose ingested before and/or during prolonged exercise. AB - Ingestion of glucose before exercise results in a transient increase in plasma insulin concentrations. We hypothesized that if glucose was also ingested during the exercise period the elevated plasma insulin concentration could increase exogenous glucose oxidation. The oxidation rate of glucose ingested 30 min before (50 g) and/or during (110 or 160 g in fractionated doses) exercise [120 min; 67.3 (1.2)% maximal O(2) uptake] was studied on six young male subjects, using (13)C labelling. Ingestion of glucose before exercise significantly increased plasma insulin concentration [from 196 (45) to 415 (57) pmol l(-1)] but the value returned to pre-exercise level within the first 30 min of exercise in spite of a continuous increase in plasma glucose concentration. Ingestion of glucose 30 min before exercise did not increase the oxidation of exogenous glucose between minutes 30 and 60 during the exercise period [0.36 (0.03) vs 0.30 (0.02) g min( 1), when placebo or unlabelled glucose was ingested respectively]. Over the last 90 min of exercise, when glucose was ingested only during exercise, 49.2 (3.1) g [0.55 (0.04) g min(-1)) was oxidized, while when it was ingested both before and during exercise, 65.7 (4.6) g [0.73 (0.05) g min(-1)] was oxidized [26.7 (2.1) g of the 50 g ingested before exercise but only 39.0 (2.4) g of the 110 g ingested during the exercise period]. Thus, ingestion of glucose 30 min before the beginning of exercise did not enhance the oxidation rate of exogenous glucose ingested during the exercise period, although the total amount of exogenous glucose oxidized was larger than when ingested only during the exercise period. PMID- 14566570 TI - Ballistic CTLA4 and IL-4 gene transfer into the lower lid prolongs orthotopic corneal graft survival in mice. AB - PURPOSE: To explore outflow from the eye and to determine and modulate the influence of lymphatic drainage on corneal graft survival in mice. METHODS: Tracer experiments were conducted in BALB/c mice using the (99m)Tc colloidal albumin Nanocoll. Count rates were determined in the eyes, submandibular lymph nodes, spleen, liver and blood 24 h after subconjunctival, intracorneal, intracameral (anterior chamber), intravenous and subcutaneous lower-lid or upper lid injections ( n=6 each). Four groups of BALB/c mice ( n=8) received corneal transplants from C3H mice; two of them were treated ballistically with vector CTLA4+IL-4 onto the leg or the lower lid, one group was untreated and the other control group was treated with an empty minimalistic, immunologically defined, gene expression (MIDGE) vector. RESULTS: Radioactivity was detected in the liver, spleen and ipsilateral submandibular lymph node after intracameral injection as follows: 91.9%, 6.6% and 1.2% respectively. Radioactivity uptake of the ipsilateral submandibular lymph node was also low after intravenous injection (0.1%) but high after intracorneal (33.8%), lower-lid (62.0%) and subconjunctival (71.2%) injection. Vector CTLA4+IL-4 treatment of the lower lid but not of the leg prolonged graft survival ( P=0.004). CONCLUSION: These tracer studies confirmed for the first time identical lymphatic drainage from the cornea and the lower lid. Logically, lymphatic drainage could be manipulated and graft survival improved by gene transfer to the lower lid. PMID- 14566571 TI - Longstanding symptomatic choroidal hemangioma managed with limited PDT as initial or salvage therapy. AB - PURPOSE: To describe the use of limited, low-irradiance, single-spot photodynamic therapy (PDT) with verteporfin for the treatment of symptomatic choroidal hemangiomas. METHODS: Thirteen consecutive patients with a circumscribed choroidal hemangioma were treated with limited PDT 6 min following a 1-min infusion with verteporfin (6 mg/m(2) body surface area), using a diode laser (692 nm) and a single spot large enough to cover only the most prominent part of the tumor. Exposure time was 166 s in the first three patients and 83 s in the last ten patients, resulting in a radiance exposure of respectively 100 and 50 J/cm(2). RESULTS: In all 11 not previously conventionally treated patients, visual acuity improved following PDT treatment. The two remaining patients with prior radiation treatment both reported widening of the visual field and sharper vision, but did not show an increase in ETDRS vision. In all but four patients the tumor became ultrasonographically undetectable by the first follow-up visit at 6 weeks, with only a slight irregularity at the level of the previous tumor remaining. In four patients the residual tumor was still detectable, and they received a second treatment, following which the tumor flattened completely. In all patients the retinal detachment disappeared. CONCLUSION: The present series demonstrates that even limited, low-irradiance, single-spot photodynamic therapy with verteporfin is an effective first-line treatment for choroidal hemangiomas. PMID- 14566572 TI - PET imaging of Rosai-Dorfman disease: correlation with histopathology and ex-vivo beta-imaging. AB - We report the case of a woman who developed an early relapse of a squamous cell carcinoma (SCC) and was thus restaged twice within a year using [(18)F]fluorodeoxyglucose (FDG) positron emission tomography (PET). While there was no evidence of metastatic tumor outspread, focally increased FDG uptake was visible in numerous nodes but showed no change during the period between the two PET scans. These nodes, predominantly located at the proximal extremities, ranged in size from about 1 cm to over 6 cm. They were located subcutaneously, showed a red/bluish livid color and were of stout consistency. These nodes occurred first after radiochemotherapy for a non-Hodgkin's lymphoma (NHL) about 6 years earlier and slowly increased in size and number. One node of the right forearm was resected and ex-vivo beta-imaging, directly measuring the positron emission of the intranodal FDG distribution, was done and showed an overall increased glucose utilization with distinct spots of high metabolism. Histopathological work-up of the tumor showed widespread granulomatous tissue with lymphocyte follicles. Immunostaining showed the tumor to be positive for S100, CD68 and vimentin. Rosai Dorfman disease (RDD) was diagnosed and no evidence of a potential relapse of the previous NHL was detected. RDD is a rare disease that is associated with the multifocal growth of benign tumors. The lesions are metabolically highly active. The correlation of the beta-imaging and histopathological results showed a high metabolism within granulomatous tissue with more intense metabolism within lymphocyte follicles. PMID- 14566573 TI - New observations of ant-dipping techniques in wild chimpanzees at Bossou, Guinea. AB - Ant-dipping behavior is often cited as a clear example of chimpanzee culture, since different populations have apparently different dipping techniques (the one handed method used by chimpanzees in Bossou, Guinea and Tai, Cote d'Ivoire, and the two-handed method used in Gombe, Tanzania). Here we report a new observation of ant-dipping behavior from Bossou using the two-handed method, in addition to the first detailed description of the one-handed method. Although the main dipping pattern was the one-handed method in Bossou, one adult male was observed dipping for ants using the two-handed method, while other chimpanzees employed the conventional one-handed method in the same episode. The two-handed method was also sporadically observed in a juvenile and in adolescents, who were still immature in dipping techniques and hence prone to suffer from ant attacks. Cross population comparisons of dipping techniques suggest that there are two sub-types of the one-handed method, and the Bossou one-handed technique may be substantially different from that of Tai. In terms of overall behavioral repertoire in ant dipping, the Bossou pattern appears more similar to that of Gombe than Tai. This may be explained by the difference in target ant species. PMID- 14566574 TI - Pulmonary aspergillosis in a patient with chronic granulomatous disease: confirmation by polymerase chain reaction and serological tests, and successful treatment with voriconazole. AB - A 25-year-old male with chronic granulomatous disease developed severe bilateral pulmonary aspergillosis confirmed by a positive result of polymerase chain reaction for Aspergillus species from sputum and positive serological tests for Aspergillus. He was successfully treated with voriconazole. The diagnostic difficulties and the new therapeutic options in the era of new antifungal agents are discussed. PMID- 14566575 TI - Nosocomial cluster of Candida guillermondii fungemia in surgical patients. AB - A nosocomial cluster of Candida guillermondii fungemia ( n=5 episodes) occurred in a surgical unit over a 2-week period. The five infected patients had received parenteral nutrition through central lines and three of them had catheter-related candidemia. All of the isolates were resistant to 5-flucytosine (MIC >32 microg/ml) and they had strictly related fingerprints, as generated by randomly amplified polymorphic DNA analysis. Although no isolate of Candida guillermondii was recovered from other clinical, surveillance or environmental samples, nosocomial spread of this yeast stopped following the reinforcement of infection control measures. Candida guillermondii may require an intravascular foreign body to cause fungemia, but the outbreak reported here shows that it can be transmitted nosocomially and cause epidemics. PMID- 14566576 TI - Brucella endocarditis: clinical, diagnostic, and therapeutic approach. AB - Brucella endocarditis is an uncommon focal complication of brucellosis. Presented here are 11 cases of Brucella endocarditis, all managed uniformly. The median duration of symptoms prior to diagnosis was 3 months. Five patients (45%) had underlying valvular damage, and in six (55%) endocarditis involved a normal valve. There was a predominance of aortic involvement (82%) and a high incidence of left ventricular failure (91%). Diagnostic suspicion was essential in order to test blood cultures correctly, which in this series were positive in 63% of the patients. Surgical treatment was undertaken in eight patients (72%), all with aortic involvement and left ventricular failure impossible to control with medication. One patient died during the immediate postoperative period. All the other patients received antibiotic therapy for 3 months, with no signs of relapse of the infection or malfunction of the prosthesis during a minimum follow-up period of 24 months. PMID- 14566577 TI - The Ai project: historical and ecological contexts. AB - This paper aims to review a long-term research project exploring the chimpanzee mind within historical and ecological contexts. The Ai project began in 1978 and was directly inspired by preceding ape-language studies conducted in Western countries. However, in contrast with the latter, it has focused on the perceptual and cognitive capabilities of chimpanzees rather than communicative skills between humans and chimpanzees. In the original setting, a single chimpanzee faced a computer-controlled apparatus and performed various kinds of matching-to sample discrimination tasks. Questions regarding the chimpanzee mind can be traced back to Wolfgang Koehler's work in the early part of the 20th century. Yet, Japan has its unique natural and cultural background: it is home to an indigenous primate species, the Japanese snow monkey. This fact has contributed to the emergence of two previous projects in the wild led by the late Kinji Imanishi and his students. First, the Koshima monkey project began in 1948 and became famous for its discovery of the cultural propagation of sweet-potato washing behavior. Second, pioneering work in Africa, starting in 1958, aimed to study great apes in their natural habitat. Thanks to the influence of these intellectual ancestors, the present author also undertook the field study of chimpanzees in the wild, focusing on tool manufacture and use. This work has demonstrated the importance of social and ecological perspectives even for the study of the mind. Combining experimental approaches with a field setting, the Ai project continues to explore cognition and behavior in chimpanzees, while its focus has shifted from the study of a single subject toward that of the community as a whole. PMID- 14566578 TI - Intellectual differences between schizophrenic patients and normal controls across the adult lifespan. AB - A debate persists about whether IQ declines during the duration of schizophrenia or whether an early deficit remains static across the lifespan. To examine this, we measured estimated current IQ (Quick Test Revised: QTR) and estimated premorbid IQ (National Adult Reading Test: NART) in schizophrenic patients (n=110) and matched healthy controls (n=71) across a wide age range (20-88). Age correlated negatively with NART and QTR IQ for schizophrenic patients, but not for controls. A subset of 23 schizophrenic patients was also retested on the NART after 4 years to determine NART stability and they showed no significant change. We propose that the lower NART IQ in older patients reflects a lower 'starting point' and that this may be related to lower educational opportunities in older patients. PMID- 14566579 TI - Effects of age, education and gender on verbal fluency. AB - The objective was to study the effects of age, education and gender on verbal fluency in cognitively unimpaired, older individuals. The methods used were as follows: cognitively unimpaired elderly (55-84 years) subjects (n=153), were administered category (animal) (CF) and letter (/pa/) (LF) fluency tasks, in their native language of Malayalam. Results and conclusions were (1) Level of education, but not age or gender, significantly influence LF. (2) Level of education (directly) and in the elderly subjects, age (inversely) affect CF. (3) Age, but not education, has a differential effect on the tasks of verbal fluency, influencing CF more than LF. PMID- 14566580 TI - RBANS performance: influences of sex and education. AB - A sample of 278 men and 353 elderly women who resided in the community received the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) as part of a study of health in mature adult Oklahomans. Females attained significantly higher scores on the Immediate and Delayed Memory and the Language Indexes. Males performed better on the Visuospatial/Construction Index, but there was no sex difference on the Attention or Total Indexes. Differences related to sex were modest (2-6 Index points), but combined with the larger effects of education could lead to an adjustment of up to 11 Index points. PMID- 14566581 TI - Semantic relations and repetition of items enhance the free recall of words by multiple sclerosis patients. AB - We compared 25 patients with multiple sclerosis (MS) and 24 normal controls on a test of free recall of words. Some lists contained words that were all unrelated, while in others the intermediary words were semantically related. In another set, the mid-list words were repeated across the lists, or, in addition to the repetition, were semantically associated. Immediate recall was assessed using these lists. Delayed recall was assessed using different lists (delay-unrelated and delay-related) after distractor tasks. Recency was not affected in MS patients, but the primacy effect was lower than in controls, this effect being interpreted as due to a deficiency in articulatory rehearsal. The delay interval after each list abolished recency in both groups and resulted in impaired recall in MS patients. However the patients, like the controls, benefited from semantic relations in the middle of the lists and from spaced repetition of words across the lists, in either immediate and delayed recall. The enhancing effects of word relatedness and of spaced repetition are seen as being due to automatic processes preserved in MS patients. PMID- 14566582 TI - Temporal preparation strategy may inflate RT deficit in patients with Parkinson's disease. AB - Twelve patients with Parkinson's disease (PD) and 12 age-matched controls completed a visual reaction time (RT) task to assess the effect of temporal parameters on response preparation. Simple and choice RT conditions were presented in separate blocks. In both conditions, preparatory intervals of various durations (1, 3 and 5 s) were introduced between an auditory warning signal and the visual target. Within a block of trials, intervals varied randomly. The results indicated that PD patients responded slower than controls in both task conditions. Also, there was evidence for preparation in both groups, as RT decreased with increasing intervals. A three-way interaction indicated that PD patients' RT was longer than that of controls at the shortest interval in simple RT. This suggests that PD patients show a different pattern of temporal response preparation and that this may contribute to their deficit on RT tasks. PMID- 14566583 TI - Social support moderates caregiver life satisfaction following traumatic brain injury. AB - Social support is an important determinant of adjustment following traumatic brain injury (TBI) sustained by a family member. The present study examined the extent to which social support moderates the influence of characteristics of the person with injury on caregiver subjective well-being. Sixty pairs of individuals who had sustained a moderate to severe TBI and their caregivers (N=120) participated. Years postinjury ranged from 0.3 to 9.9 ( M=4.8, SD=2.6). Cognitive, functional, and neurobehavioral functioning of participants with TBI were assessed using neuropsychological tests and rating scales. Caregiver life satisfaction and perceived social support were assessed using self-report questionnaires. Results indicated that time since injury was unrelated to life satisfaction. Neurobehavioral disturbances showed an inverse relation with life satisfaction. Social support emerged as an important moderator of life satisfaction. Only among caregivers with low social support was cognitive dysfunction adversely related to life satisfaction. Similarly, a trend suggested that patient unawareness of deficit was associated with caregiver life dissatisfaction only among caregivers with low social support. In contrast, these characteristics were unrelated to life satisfaction among caregivers with adequate social support. PMID- 14566584 TI - Environmental influences on primary memory development: a cross-cultural study of memory span in Lao and American children. AB - Although research examining Auditory-Verbal and Visual Memory Span in adults has failed to provide compelling evidence for the modality specificity of such tasks, Fastenau, Conant, and Lauer (1998) suggested that these processes may be dissociable in children until around the age of 8 years when visuospatial memory span appears to become more verbally mediated perhaps as a function of formal reading training. A subsequent cross-cultural study in Zaire (now Democratic Republic of Congo) by Conant and colleagues (1999) supported that hypothesis. However, other differences in the sociocultural environments of American and rural Zairian children such as nutritional and health care factors could potentially account for the findings. The present study examines the relationships among Auditory-Verbal, Visual, and Cross-Modal Memory Span tasks with 46 Lao and 56 American children. Both samples were divided into younger and older groups, and separate partial correlational analyses were performed on Kaufman Assessment Battery for Children (K-ABC) data from each group. In younger Lao and American children, Auditory-Verbal Memory Span was not significantly correlated with Visual Memory Span or Cross-Modal Memory Span, whereas significant correlations were found between Auditory-Verbal Memory Span and these other tasks in older children from both countries. The similar pattern in both cultures, together with findings from Zaire, supports the hypothesis that the relationship between Auditory-Verbal and Visual Memory Span may change with reading training. PMID- 14566585 TI - A controlled quantitative MRI volumetric investigation of hippocampal contributions to immediate and delayed memory performance. AB - MRI volumetric TLE studies show inconsistent evidence of hippocampal involvement in memory. Prior studies have not dissociated hippocampal and temporal lobe contributions to memory. We measured hippocampal and temporal lobe volumes and immediate/delayed memory performances in 64 TLE patients. Regression was used to dissociate hippocampal from temporal lobe contributions to memory. Results revealed reliable evidence for dominant hippocampal involvement in delayed verbal recall across three separate measures and less consistent evidence for nondominant hippocampal involvement. The findings point to a consistent relationship of dominant hippocampal volumes to delayed verbal recall but no involvement of the temporal lobe or nondominant hippocampus in memory. PMID- 14566586 TI - Quantifying dissociations in neuropsychological research. AB - Double dissociations play an important role in neuropsychology, but they are often identified through subjective estimates of "high" versus "low" performance, without considering the probability that such an outcome might have occurred by chance. To determine whether two measures "come apart" in an interesting way in brain-damaged patients, it is important to know the degree to which variance in one measure can be predicted by variance in the other. This study introduces a statistical procedure to determine the probability of a double dissociation when the correlation between measures is taken into account. Different quantitative definitions of dissociations were compared in two large samples of neurological patients, and applied to four pairs of measures (two for language, two for hemispatial neglect) with different degrees of intercorrelation (ranging from +.21 to +.84). If the correlation between measures is not taken into account, large numbers of dissociated cases may be missed, especially for measures that are highly correlated. There are also qualitative differences between methods in the identity of those individuals who meet each definition. PMID- 14566587 TI - Neglect and hemianopia superimposed. AB - In patients with posterior-parietal brain damage it is often difficult to decide whether left-sided omissions in perimetry are due to primary visual loss or due to visual neglect. We investigated 11 patients with combined neglect/hemianopia and 11 patients with pure hemianopia using a visual search task with single or double stimulation conditions. The second stimulus was either the fixation point itself (like in perimetry) or a distractor appearing in the hemifield opposite to the target. The fixation point did not worsen left-sided perception, but its disappearance led to a bias of exploration towards the right side in neglect patients but not in pure hemianopics. A distractor in the intact hemifield worsened the performance to left-sided stimuli, that is, neglect patients behaved as if they were completely hemianopic, even in intact parts of the visual field (VF). Three of the neglect patients showed unconscious processing of the distractor in the left VF, suggesting that the visual field defect was produced by neglect mechanisms rather than primary visual loss. This visual search paradigm appears to be helpful in understanding of the nature of hemianopia versus neglect deficits in individual patients. PMID- 14566588 TI - A 3D virtual environment rod and frame test: the reliability and validity of four traditional scoring methods for older adults. AB - This study introduced a 3D rod-and-frame test in a virtual environment (VERF) and evaluated its reliability and concurrent validity according to four scoring methods previously used with 2D tests. Thirty nonimpaired older adults (15 men; 15 women) aged 64-86 (M=73.6) manipulated a rod in a semi-immersive 3D virtual environment. Results suggested that the conventional 2D scoring method of absolute deviation from vertical was the most reliable method (alpha=.79; gamma=.89). This approach also had higher correlations with traditional neuropsychological measures of spatial ability than three other methods. Given the availability of a reliable and valid scoring method, the VERF may result in previously unavailable insights through the measurement of visuospatial abilities in 3D. PMID- 14566589 TI - Rated familiarity, visual complexity, and image agreement and their relation to naming difficulty for items from the Boston naming test. AB - The purpose of this experiment was to obtain data on the name agreement, familiarity, visual complexity, and imageability of the 60 line drawings of objects that comprise the Boston Naming Test (BNT). One group (N=64) named and then rated each item for visual complexity. Another group (N=98) rated the familiarity of each depicted concept and a third group (N=41) rated each drawing for how closely it matched their mental image of the item's name. Significant correlations between several measures were found and the results were consistent with those obtained by Snodgrass and Vanderwart (1980) for 21 overlapping concepts depicted in both the BNT and their set of items. PMID- 14566593 TI - VXII International Symposium on Morphological Sciences. Proceedings of the minisymposium on salivary glands. September 11-15, 2002. Timisoara, Romania. PMID- 14566590 TI - Neurocognitive functioning is associated with employment status: a quantitative review. AB - Neurocognitive assessment is frequently used as a basis for making determinations regarding a person's ability to work; yet, to our knowledge, a review of the association between neurocognition and employment status has not been conducted. For this review, we utilized meta-analysis to quantify objectively the association between eight neurocognitive domains and employment status. The meta analysis revealed that performance in each domain was significantly associated with employment status, and that the associations were greatest for the following domains: intellectual functioning, executive functioning, and memory. These findings support the ecological validity of neurocognitive assessment. PMID- 14566596 TI - Relationship of fluid and mucin secretion to morphological changes in the perfused rat submandibular gland. AB - Transport of electrolytes/water and exocytosis are activated by elevation of the cytosolic Ca(2+) concentration and are potentiated by elevation of cytosolic cyclic AMP. To correlate mucin and fluid secretion with morphological changes, rat submandibular glands were vascularly perfused and the fluid secretion and N acetylgalactosamine in the saliva were measured during stimulation with various concentrations of carbachol (CCh) and/or isoproterenol (ISP). Single stimulation with 1 microM CCh induced a transient increase of N-acetyl galactosamine followed by a decline to a low level during sustained stimulation. The overload of 1 microM ISP increased secretion of N-acetyl galactosamine to a higher sustained level of 40-50 microg/g-min. However, at 1 microM CCh, fluid secretion was maintained at the same level during stimulation and even overload of 1 microM ISP did not significantly affect its level, whereas addition of 0.5 microM ISP to the gland stimulated with 0.1 microM CCh increased fluid secretion. Morphological observation was carried out by HRSEM and TEM. Combination of CCh and ISP in different concentrations resulted in distinctive morphological changes which reflect fluid secretion and mucin secretion. The kinetics of ATP and creatine phosphate (PCr) were measured using P-31 NMR, which indicated that the potentiation of fluid secretion is limited under a higher level of CCh stimulation due to a limited energy supply. PMID- 14566597 TI - Expression and distribution of osteopontin and matrix metalloproteinase (MMP)-3 and -7 in mouse salivary glands. AB - We investigated the expression and distribution of osteopontin in mouse salivary glands. Western blot analysis showed intense positive bands at the predicted molecular mass (about 60 kDa) in mouse parotid and sublingual glands. However, a cross-reacted band around 30 kDa was strongly detected in submandibular glands. Indirect immunofluorescent analysis showed that osteopontin was localized at the luminal (apical) membranes of the acinar cells in parotid and sublingual glands. However, it was not detected in acinar cells of submandibular glands. No expression was found in ductal cells of any glands. We also examined the expression of matrix metalloproteinase (MMP)-3 and -7. In parotid gland, MMP-3 was observed at 57 kDa, indicating a latent form, but MMP-7 was not detected. In contrast, MMP-7 definitely was observed at 28 kDa area in submandibular gland, whereas MMP-3 was not detected. These results suggest that osteopontin localizes at luminal sites of acinar cells and may be associated with saliva secretion in mouse salivary gland. It is also suggested that osteopontin may be cleaved by MMP 7 in mouse submandibular gland. PMID- 14566598 TI - Ultrastructural localization of epidermal growth factor receptor in human parotid gland. AB - The epidermal growth factor receptor (EGFR) is widely distributed in several organs in which, following interaction with its ligand, it can affect development and differentiation. The aim of this study was to define the distribution of EGFR in human parotid gland by means of a post-embedding immunogold staining method. Normal human parotid glands obtained at surgery were routinely prepared for electron microscopy. Semithin and ultrathin sections were treated for immunocytochemistry using a mouse monoclonal antibody specific for EGFR and a goat anti-mouse gold conjugated secondary antiserum. At the light microscope level, EGFR reactivity was revealed by a specific dark staining in both acinar and ductal cells. At the electron microscope level, EGFR was strongly stained in the cytoplasmic compartments and occasionally labeled on cell surfaces. In acinar cells, it appeared to be associated with small vesicles of uncertain nature that were scattered among the secretory granules. EGFR-positive vesicles were also observed in the ductal cells, with the most intense labeling being localized in striated ducts. Since cytoplasmic vesicles were previously found to be EGF positive, these results may be due to the presence of the EGF-EGFR complex that is internalized after binding of EGF to the surface EGFR. PMID- 14566599 TI - Cyclic AMP-receptor proteins in human salivary glands. AB - In mammalian species, cyclic AMP receptor proteins (cARP) are the regulatory (R) subunits of cyclic AMP-dependent protein kinase (PKA), the cellular effector of cyclic AMP-mediated signal transduction. An isoform of the PKA type II R subunit (RII), cARP, is a polyfunctional protein, present in most tissues and cells. It is expressed in salivary and other glands of rodents, and secreted into the saliva of rats and Man. The aim of the present study was to determine the expression of cARP in human salivary glands using immunoelectron microscopy. Thin sections of normal salivary glands embedded in LR Gold resin were labeled with anti-cARP primary antibody, then with gold-conjugated secondary antibody. Labeling was present in the secretory granules and cytoplasm of parotid, submandibular (SMG) and sublingual gland serous cells. Quantitative analysis showed considerable variability in granule labeling from sample to sample, indicating shifts in expression and cellular location of cARP. Unlike rodent salivary glands, the granules of intercalated and striated duct cells also were labeled. The cytoplasm and granules of mucous cells of the SMG and sublingual glands were unlabeled, while the Golgi complex and filamentous bodies in these cells showed moderate reactivity. Mitochondria and nuclei of both serous and mucous cells were unlabeled. Labeling also was present in the connective tissue adjacent to the epithelial cells. The results indicate that serous cells of the parotid and SMG are the major source of salivary cARP. They also reveal significant species differences in the glandular distribution of RII. RII binds to cytoskeletal and nuclear proteins, and may function to regulate extracellular cyclic AMP levels. Thus, the tissue and cellular distribution of RII may serve as an index of regulation of gene expression and cell differentiation. PMID- 14566601 TI - Three-dimensional structure of apical vesicles of tuft cells in the main excretory duct of the rat submandibular gland. AB - Tuft cells are present in the mucosal epithelium of a number of hollow organs including the main excretory duct (MED). Despite their distinctive features such as the long, thick, blunt microvilli with prominent rootlets and the large number of vesicles in the apical cytoplasm, the hypolemmal terminal-tuft cell relationship and the true form of the various vesicles and tubules are still controversial. The present study investigated the above mentioned features of tuft cells in the MED of rat submandibular gland by computer three-dimensional (3 D) reconstruction with focus on their function. Computer 3-D reconstruction revealed that nerve endings are present at both sides of the basal portion of the lateral cytoplasmic branch of tufts cells and that the apical tubulovesicular system of these cells consists of two separate components: the complex and coherent vesicles and the small network of tubules. We suggest that such a system may be involved in the rapid changes of surface area observed in tuft cells. Furthermore, our findings demonstrate that the images seen in thin sections and formerly regarded as evidence for the presence of variations in the shape of the tubules and of the vesicles are in reality the product of the different angles at which the tubulovesicular system was sectioned. Finally, a few vesicles and tubules that were not part either of the complex or of the network, also were found. PMID- 14566600 TI - Morphofunctional studies on human labial salivary glands. AB - In this study, the first experimental investigation carried out at the ultrastructural level on mucous cells of human salivary glands, we have examined by light microscopy (LM), transmission electron microscopy (TEM), high resolution scanning electron microscopy (HRSEM), the secretory response of labial glands stimulated in vitro by the beta-adrenergic agent, D,L isoproterenol, and by the muscarinic agent carbachol. For comparison we have used identical methods to study samples of mixed portions of human submandibular glands. Morphological findings obtained here on both submandibular and labial glands mucous cells demonstrate that mucous droplets are released solely by muscarinic stimulation, and that cytological events occurring during secretory discharge are similar to those described by others, using TEM, on stimulated mucous cells of rat sublingual glands. Despite the fact that human labial glands are said to have a prominent cholinergic innervation with scanty adrenergic nerves, the response of seromucous cells in these organs to stimulation with carbachol and with isoproterenol was similar to that observed by us, (using LM, TEM and HRSEM), in serous cells of human major salivary glands. PMID- 14566602 TI - Visualization of 'water secretion' by confocal microscopy in rat salivary glands: possible distinction of para- and transcellular pathway. AB - Visualization of water transport in cells, tissues and organs is an important, yet still difficult, task in morphological science. By using confocal microscopy and the fluid-phase fluorescent tracer technique, we visualized water secretion and estimated the routes of water transport across the acinar epithelia in rat parotid and submandibular glands. Confocal microscopy of whole glands perfused arterially with Lucifer yellow revealed a bright fluorescence at the basolateral space of acini. Luminal space was devoid of fluorescence, but revealed it after isoproterenol pretreatment, ductal infusion of fluorescent dextrans into the lumen, or tissue dissociation by collagenase. Under these conditions, stimulation of fluid secretion with carbachol caused a rapid decline of the luminal fluorescence intensity, indicating that the secreted water washed out the fluorescent probes in the acinar lumen. In the stimulated dissociated acini, the luminal fluorescence disappeared by 15 sec, but reappeared at 30-45 sec to maintain a low plateau level. By assuming that the tight junction was 'paralyzed' by the collagenase digestion and that the paracellular fluid transport could not influence the dilution of Lucifer yellow, we estimated that the initial water secretion by CCh occurs via the transcellular pathway, while later than 30-45 sec the additional water permeates through the paracellular pathway. PMID- 14566603 TI - Buds of the Golgi apparatus in parotid acinar cells. AB - The Golgi apparatus (GA) is a membranous organelle composed of stacked cisterns with associated vesicles. This study was undertaken to determine its origin in rat parotid acinar cells. The morphogenesis of the GA could be recognized in the developmental process as well as in mitotic division of cells. EM studies depicted an aggregation of small vesicles in the early stage of postnatal development or mitosis, that appeared to be the rudimental element of GA. Brefeldin A induced rapid degradation of the cisternal structure to vesicular aggregates. Reconstruction of the GA structure based on these remnant vesicles was observed upon removal of the drug. Similar membranous assembly could be observed after destruction of microtubules. These membranous aggregates presumably corresponded to 'buds of the GA' in parotid acinar cells. However, conventional cytochemical markers for GA were not detected on such immature form of GA. We found that the GA matrix protein GM130 and osmium reductivity (a classical marker for cis-Golgi elements) were consistently localized in the GA elements. Therefore, immunohistochemical distribution of GM130 and osmium impregnation of parotid acinar cells were studied under various dynamic conditions that produced structural modification of the GA. PMID- 14566604 TI - Ultrastructure of the submandibular gland of the rare white-winged vampire bat, Diaemus youngi. AB - The submandibular gland of the white-winged vampire bat, Diaemus youngi, was examined by electron microscopy. Unlike typical submandibular glands, those in Diaemus have only one type of secretory cell in their endpieces, namely, serous cells. These serous cells are conventional in structure, with an extensive rough endoplasmic reticulum, scattered dictyosomes, and numerous secretory granules. The endpiece lumina, as well as intercellular canaliculi, are fitted with numerous microvilli, which also are present on the otherwise unremarkable intercalated duct cells. Striated ducts are of conventional morphology, but have a brush border-like array of microvilli on their luminal surface. These cells resemble those in the submandibular gland of the common vampire bat, Desmodus rotundus. The presence of an abundance of microvilli in the salivary glands in the two vampire bat species (and their absence from chiropteran species that consume other types of diets) is a strong indication that these structures play a significant role in dealing with the problems posed by a sanguivorous diet. PMID- 14566606 TI - Morphological and morphometrical characteristics of the esophageal intrinsic nervous system in the golden hamster. AB - Very little is known about esophageal innervation in the hamster. In the present study, we used protein gene product 9.5 (PGP 9.5) to determine immunohistochemically the architectural features of the enteric nervous system in the hamster esophagus. The myenteric plexus consisted of a loose and irregular network of ganglia and interganglionic nerve bundles. The density of the neurons in the myenteric plexus was relatively low (479 +/- 75/cm(2), n = 5), with a preferentially higher density in the upper cervical portion than other parts of the esophagus. Regional differences in the number of PGP 9.5-positive neurons and ganglia were observed. PGP 9.5-immunoreactive fibers in the ganglia often branched, giving rise to expanding nerve endings of laminar morphology resembling intraganglionic laminar endings described in rats and cats. Fine varicose fibers originating from the secondary plexus were occasionally observed near the motor endplates, suggested a dual innervation of the striated muscle. The submucosal plexus was free from ganglionated plexus. A regional difference in the submucosal nervous network was observed. The number of motor endplates in the inner muscle layer was higher than that in the outer muscle layer. PMID- 14566605 TI - Gene expression, cell localization, and evolution of rodent submandibular gland androgen-binding protein. AB - A small dimeric androgen-binding protein (Abp) secreted by mouse (Mus) submandibular salivary glands has been hypothesized to function in mate selection. The alpha-subunit (Abpa) evolves rapidly under natural selection. However, cellular site(s) of synthesis, mode of function, and patterns of evolution of this biologically important protein are otherwise unknown. We used a radiolabeled riboprobe and in situ hybridization to mouse (Mus) Abpa mRNA to localize Abpa synthesis to submandibular gland acinar cells. We next used a quantitative fluorescent thermal cycler (real-time PCR) to determine relative expression levels (normalized with a constitutively expressed ribosomal gene [S15/rig; rat insulinoma gene]) in male, female, and sexually immature European striped field mice (Apodemus agrarius). We hypothesized that gender or age related differences might occur in production of a salivary protein related to sexual selection, but found no significant differences within our sample. Finally, we also used reverse transcription-PCR of mRNAs isolated from submandibular salivary glands to determine the Abpa allele in the striped wood mouse and compare it to published information on the homologue in Mus musculus domesticus. PMID- 14566607 TI - Experimental analysis of error sources in fibre type counts of biopsies in horses. AB - The contribution to total variance of different error sources in fibre type counts of equine gluteus medius muscle biopsies was determined to quantify and possibly improve the resolution of the method. Fibre types were defined on the basis of myosin heavy chain immunostaining. Errors were determined at levels: (1) positioning the insertion channel, (2) positioning the needle tip (3) biopsy heterogeneity (4) observer interpretation. Errors at levels 1 and 2 were considerable. Confidence intervals for individual observations were +/- 10-15%. In longitudinal studies a group size of 4 animals is necessary to resolve fibre composition changes of 10%. Comparison with multiple counts on post mortem specimens showed that local muscle fibre heterogeneity is responsible to a considerable extent for the error variance. Variance is effectively reduced by processing 3-4 shavings from the same insertion channel. PMID- 14566608 TI - Absolute numbers versus proportions in the assessment of differentiation of beta cells in the embryonic avian pancreas. AB - Environmental factors may influence the proliferation and differentiation of embryonic pancreatic endocrine cells, creating a need for the quantification of such effects. The explanted dorsal pancreatic bud (DPB) of the 5-day chick embryo is a useful in vitro model. Since all explants cannot be assumed to have the same number of endocrine cells at the start of culture, the proportion of beta-cells with respect to alpha-cells may be a more meaningful measure than absolute numbers. This study aimed to establish baseline values for the proportion of beta cells in both intact and mesoderm-depleted DPBs before culture. Buds were excised from 12 chick embryos and the surrounding mesoderm was removed from 6 buds following collagenase treatment. All the buds were freeze-dried, fixed in parabenzoquinone vapour, embedded in resin and sectioned at 1 micro m. alpha- and beta-cells were detected by an indirect immunoenzyme method. alpha-cells outnumbered beta-cells in 9 of the 12 buds. The proportion of beta-cells in the intact buds varied from 16% to 64% (mean 39.5%) and in the mesoderm-depleted buds from 17% to 66% (mean 39%). There was no significant difference between the absolute numbers or the proportions of cells in either case. The proportions of beta-cells in the 5-day DPBs were higher than those in buds cultured in previous studies for 7 days under various conditions. This result may reflect the role of apoptosis in response to the culture conditions. PMID- 14566609 TI - Scanning electron microscopic study of the structures associated with lips of an Indian hill stream fish Garra lamta (Cyprinidae, Cypriniformes). AB - The surface architecture of the structures associated with the lips of a hill stream fish Garra lamta was examined by scanning electron microscopy. In this teleost, the lips are inconspicuous and associated with prominent horny jaw sheaths. Furthermore, the upper and lower lips are associated with a greatly enlarged rostral cap and an adhesive pad, respectively. The rostral cap has a proximal mucogenic region and a distal keratinized region. The adhesive pad is differentiated into central mucogenic and peripheral keratinized regions. At the mucogenic regions of the rostral cap and the adhesive pad, the surface of the epithelial cells is characterised by well developed microridges, which reflect their high secretory activity. The mucus may lubricate the surface and protect the epithelia from abrasions at these regions, which are likely to be subjected to wear and tear during frequent friction and adhesion of the fish to the substrate. Taste buds may help the fish to locate food and trigger a 'pick-up' reflex. The epithelial cells at the surface of the keratinized regions of the rostral cap and the adhesive pad are modified as clusters of spine like unculi, which may assist the fish in its firm anchorage to the substrate. The epithelial cells at the surface of the horny jaw sheaths are modified as polygonal unculi, each appears much like a tooth that has a characteristic sharp edge at the margin. These may be regarded as an adaptation to browsing or scraping food materials from the substrate. PMID- 14566610 TI - The incidence of basal sphenoid bony bridges in dried crania and cadavers: their anthropological and clinical relevance. AB - The aim of this study was to investigate the incidence of the pterygospinous and pterygoalar bony bridges and the variations in these bony bridges among Anatolians. A total of 452 adult dry skulls (258 males and 194 females) of the Anatolian population were investigated for both the pterygospinous and the pterygoalar bony bridges. In 80 of the 452 dry skulls (37 male and 43 female), it was possible to inspect the cranial cavity. In these skulls, sellar and sphenopetrous bridges were also investigated. In addition to this, the mandibular nerve of 9 fixed cadavers was carefully dissected and the distribution of its branches was determined on both sides. Complete pterygospinous osseous bridges were found in 5.5% of the samples and complete pterygoalar bridges in 4.9%. In the dry skulls with removed calvaria, complete sellar osseous bridges were found on both sides in 34.2% of specimens, complete pterygospinous bridges in 8.8% and complete pterygoalar bridges in 7.5%. No complete sphenopetrous bridges were found. In the cadaveric study, nerve entrapment due to a pterygoalar ligament on the left side was found in one cadaver. Such variations should be kept in mind in clinical complaints such as mandibular neuralgia, especially during chewing. PMID- 14566611 TI - New aspects of the topographical anatomy of the mammary gland regarding its neurovascular supply along a regular ligamentous suspension. AB - The exact location of the main nerves and vessels to the breast and the nipple areola complex has always been obscure. We found that the course of the rich neurovascular supply to the nipple runs along a regularly-located, suspensory apparatus and can therefore be predicted exactly. It consists of a horizontal fibrous septum originating at the pectoral fascia along the 5th rib, merging into vertical ligaments along the sternum medially and along the lateral border of pectoralis minor laterally. Cranially, and in an anterior direction, the vertical ligaments are connected by the superficial fascia. In the current anatomical study, we seek to demonstrate the vascular supply provided by these structures more impressively. For this purpose we dissected the ligamentous suspension after intraarterial injection with colored latex in both breasts of 10 female cadavers. The large vessels, guided by this circle of fibrous attachments could then be seen clearly. In a further 4 female cadavers, a similar procedure was performed after intraarterial injection of surgical ink. This stained the vascular layers even more intensely. This topographical knowledge has clinical relevance. The rich and constant neurovascular supply to the nipple areola complex may be maintained in a new breast-reduction technique, which allows safe postoperative viability and sensibility of the nipple. The clinical results act as a striking evidence of our anatomical findings. Further procedures taking advantage of the easy determination and access to the neurovascular supply may be seen in future. PMID- 14566623 TI - Correlation between morphological and functional retinal impairment in patients affected by ocular hypertension, glaucoma, demyelinating optic neuritis and Alzheimer's disease. AB - In this article the correlations between the morphological evaluation of the nerve fiber layer (NFL) thickness (by OCT) and retinal functional assessment (by Pattern ERG recordings) performed in patients affected by ocular hypertension (OHT), glaucoma (OAG), demyelinating optic neuritis (MSON), and Alzheimer's disease (AD) are reported. In OHT eyes with ocular hypertension we observed that the inter-individual variation in NFL thickness is correlated with the variability of the PERG responses (the thinner the layer, the worse the visual function). In our OAG, MSON and AD eyes we observed a significant reduction in NFL thickness when compared with controls. In OHT, OAG, MSON and AD eyes abnormal PERG responses with delayed implicit times and reduced amplitudes were found. The impairment in the PERG parameters was significantly correlated to the reduction in NFL thickness. Our results suggest that in patients affected by ocular hypertension, glaucoma, demyelinating optic neuritis, and Alzheimer's Disease there is a reduction of NFL thickness evaluated "in vivo" by OCT, and this morphological involvement is correlated with electrophysiological responses assumed to be originating from the innermost retinal layers. PMID- 14566624 TI - Macular hole: improved understanding of pathogenesis, staging, and management based on optical coherence tomography. AB - Optical coherence tomography (OCT) has improved the understanding and management of idiopathic macular hole. Images of precursor lesions and their progression demonstrate that antero-posterior as well as tangential traction are involved in macular hole formation. The staging of macular holes with biomicroscopic examination can be paralleled by optical coherence tomography staging with some modification. OCT is useful in differentiating simulating lesions and in allowing better counseling of patients regarding their disorder. Finally, OCT findings are prognostic of surgical success and can assist in evaluating the results of surgery. PMID- 14566625 TI - Treatment of central retinal vein occlusion with triamcinolone acetonide: an optical coherence tomography study. AB - Central retinal vein occlusion is one of the most common retinal vascular disorders. Many patients have decreased visual acuity as a result of macular edema. We report a retrospective review of 8 patients at the University of Wisconsin with macular edema from CRVO who were treated with an intravitreal injection of triamcinolone acetonide. Optical coherence tomography (OCT) was used to help assess the effect of this intervention. Mean baseline visual acuity was 20/500. Mean visual acuity at the 3-month follow up was 20/220. The average gain in visual acuity was 3.3 lines (range -1 to +10). Four of 8 patients experienced a visual acuity gain of 2 or more lines at the 3-month follow up. Four of 8 patients were unchanged (within 2 lines of baseline) at the 3-month follow up. No patient had a decrease in visual acuity (2 or more line decrease from baseline). Seven of 8 patients had complete resolution of macular edema on clinical examination at the 3-month follow up. No adverse effects such as cataract, glaucoma, retinal detachment or endophthalmitis were noted. We conclude that intravitreal injection of triamcinolone acetonide may be a safe and effective treatment in some patients with macular edema due to CRVO. Optical coherence tomography demonstrated significant anatomic improvement in the majority of patients with macular edema due to CRVO treated with intravitreal injection of triamcinolone acetonide. PMID- 14566626 TI - Role of OCT in the diagnosis and follow up of diabetic macular edema. AB - The aim is to present, along with a brief literature review, the results of OCT scan in eyes with diabetic macular edema (DME), as well as examples of the utility of OCT for different therapeutic approaches. One-hundred and thirty-six eyes with diabetic retinopathy were analyzed with OCT to explore the different patterns of DME. Some eyes with DME were studied with OCT pre and postoperatively to determine the efficacy of photocoagulation and vitrectomy to restore a normal macular profile. Sixty-eight eyes with a central foveal thickness of 200 mu or more were considered "edematous". Three different patterns of DME were recognized and analyzed: macular thickening, cystoid macular edema and shallow retinal detachment. The change in macular profile and internal retinal structure after laser or surgical treatment are well visible with OCT. OCT contributes in understanding the anatomy of DME and the intraretinal damage and seems to be the technique of choice for the follow-up of macular edema. We think that this tool should always be used in monitoring the effect of therapies in future studies. PMID- 14566627 TI - OCT in the monitoring of visual recovery after uneventful retinal detachment surgery. AB - The aim is to describe 5 cases of unexplained visual impairment after uneventful episcleral surgery associated with submacular lesions occurring post-operatively (bleb-like syndrome). The 5 cases were followed with biomicroscopy and OCT for up to 6 months. At biomicroscopy the 5 eyes presented no visible abnormalities or a yellow subfoveal dot. OCT demonstrated the presence of isolated or multiple subfoveal or submacular blebs. Some cases experienced partial or total slow spontaneous resolution, while other blebs remained stable for months or slightly enlarged. In conclusion, OCT can play an important role in the follow up of patients with unexplained visual impairment after episcleral surgery and diagnose the presence of small subfoveal shallow retinal detachments persistent for months after surgery. We think that OCT should be routinely considered in the follow-up of these patients. PMID- 14566628 TI - Optical coherence tomography in the evaluation of vitreoretinal disorders of the macula in highly myopic eyes. AB - A higher incidence of vitreoretinal disorders is found in highly myopic eyes; however, in the same eyes, several factors contribute to make visualization of the macular area most difficult. As a consequence, vitreoretinal macular disorders are often misdiagnosed in these eyes. Ocular coherence tomography (OCT) has proven to be an invaluable technique for the evaluation of the microscopic features of the macula in highly myopic eyes, allowing us to identify which patients are at higher risk of complications, before surgical or parasurgical procedures are embarked upon. Several macular disorders, characteristic of highly myopic eyes, are described. PMID- 14566630 TI - Phenotypical characteristics, lifestyle, social class and uveal melanoma. AB - PURPOSE: To investigate potential risk factors of uveal melanoma, including phenotypical characteristics, eye burns, social class, smoking and alcohol consumption. METHODS: A hospital-based and population-based case-control study of uveal melanoma was carried out from 1995 through 1998 and the results pooled. A total of 118 patients (59 men and 59 women) with uveal melanoma and 475 controls matching on sex, age and study regions were interviewed. Conditional logistic regression was used to calculate odds ratios (OR) and 95% confidence intervals (95% CI). RESULTS: There was an elevated risk for blue or grey iris color (OR = 3.0, 95% CI 1.5-6.0). Red or blond hair color at age 20 was slightly associated with an increased risk for uveal melanoma (OR = 1.5, 95% CI 0.9-2.4). There was no elevated risk for a history of eye burns (OR = 1.1, 95% CI 0.5-2.4). CONCLUSIONS: Among the potential risk factors studied, only the phenotypical characteristics showed an association with the risk of uveal melanoma. PMID- 14566631 TI - The impact of cataract surgery on depression among older adults. AB - CONTEXT: Aged-related cataract is the leading cause of vision impairment in the elderly. Elderly individuals with cataract not only suffer from the difficulties in daily activities, but also are more prone to depression. OBJECTIVE: To examine the impact of cataract surgery on depression among older adults. DESIGN: Longitudinal follow-up study. SETTING: Outpatient ophthalmology and optometry clinics in Birmingham, Alabama. PATIENTS: Potential subjects were identified through consecutive chart review of patients seen in 10 ophthalmology and 2 optometry clinics. Three groups of individuals were identified: cataract patients who underwent surgery, cataract patients who did not undergo surgery, and patients without cataract. MAIN OUTCOME MEASURE: Depressive symptoms as measured by the Center for Epidemiological Studies-Depression Scale (CES-D). RESULTS: There was no significant difference between the baseline and follow-up CES-D scores within each group. The unadjusted CES-D score changes did not differ significantly among the three groups. Adjustment for visual acuity and contrast sensitivity in the better and worse eyes, co-morbid conditions, age, gender, and education did not alter this pattern of results. CONCLUSIONS: Cataract surgery does not appear to have an effect on reducing depressive symptoms in elderly people. PMID- 14566632 TI - Twin study of age-related macular degeneration. AB - PURPOSE: To evaluate the genetic contribution in age-related macular degeneration (ARMD) by a disease-ascertained twin study. METHODS: Concordance rates for ARMD in 25 twins were obtained by using four masked graders to confirm the diagnosis of ARMD and place subjects in one of three categories; concordant, intermediate, or discordant. Demographic features and known risk factors for ARMD were compared between monozygotic and dizygotic twin pairs. RESULTS: Of the 25 twin pairs, 15 were monzygotic and 10 were dizygotic. All 15 monozygotic twins were concordant or intermediate for ARMD. Of the dizygotic twin pairs, only one was concordant and five were discordant. In the demographic and risk factor analysis no unusual contributing or confounding variables were detected. CONCLUSIONS: The association between zygosity and concordance for ARMD suggests a major importance for genetics in the etiology of ARMD. Our data further support a multi-factorial, primarily polygenic etiology for the condition. PMID- 14566633 TI - Prevalence and causes of blindness in Otibhor Okhae Teaching Hospital, Irrua, Edo State, Nigeria. AB - PURPOSE: This hospital-based retrospective study was aimed at providing baseline information on the causes of blindness in the locality. METHODS: The case notes of all new patients attending the Eye Clinic of Otibhor Okhae Teaching Hospital, Irrua, Edo State, Nigeria, over a six-year period (January 1995-December 2000) were retrieved and analyzed. RESULTS: Over 6% (555) of new patients seen during this period were uniocularly blind while 3.9% (354) were binocularly blind. The leading causes of uniocular blindness were cataract, open-angle glaucoma and corneal ulceration/leucoma. Binocular blindness was mainly due to cataract, open angle glaucoma and aphakia. CONCLUSIONS: The prevalence of blindness in the study population is high. Cataract, as the main cause of blindness, will require surgical relief, either in the teaching hospital or preferably in the patient's locality. Appropriate interventions need to be evolved (in the form of either mobile clinics or a series of surgical eye camps) to stem the present trend toward high prevalence of avoidable blindness. Prevention of ocular trauma is an essential factor in the reduction of blindness in children. Health education and bringing ophthalmological care to the doorstep of underprivileged rural dwellers will improve their level of awareness. PMID- 14566634 TI - Asteroid hyalosis in an older population: the Blue Mountains Eye Study. AB - PURPOSE: To assess the prevalence of and associations with asteroid hyalosis in an older population. METHODS: The Blue Mountains Eye Study was a cross-sectional study of an older community (aged 49-97 years). Subjects included were those attending the baseline (n = 3654) and 5-year examinations (n = 2335) of this cohort. Asteroid hyalosis was diagnosed clinically by the presence of cream-white spherical bodies within the vitreous or from grading of stereo retinal photographs of both eyes. Logistic regression assessed age-sex adjusted associations with relevant characteristics, including diabetes and cardiovascular variables. RESULTS: Asteroid hyalosis was found in 36 subjects (1.0%), 95% confidence interval (CI) 0.7% to 1.3%, and was bilateral in three affected subjects (8.3%). An age-related increase in prevalence was observed, increasing from 0% of persons aged less than 55 years to 2.1% of persons aged 75 years or older. The prevalence of this sign was significantly higher in men (1.4%) than in women (0.6%), the age-adjusted odds ratio (OR) was 2.54 (CI 1.25-5.16). No statistically significant associations were found between asteroid hyalosis and a history of heart disease, gout, current smoking, the highest level of alcohol consumption or with presence of diabetes (diagnosed from history or fasting blood glucose tests). CONCLUSIONS: Asteroid hyalosis was detected in 1% of participants in this Australian older population. No significant associations were found, apart from age and male gender. Our study provides similar age-specific prevalence data to a recent report from the Beaver Dam Eye Study for asteroid hyalosis. PMID- 14566635 TI - Methods and design of the Chennai Glaucoma Study. AB - PURPOSE: To describe the methodology of a population-based study to estimate the prevalence of glaucoma in a rural and urban South Indian population and to study the genetics of glaucoma in this population. METHODS: A sample size of 4758 each for rural and urban populations in the Indian state of Tamil Nadu was calculated. Eligible subjects aged 40 years and above from the rural study area covering 32 contiguous villages and the urban area comprising five random clusters in Chennai city are enumerated. Demographic data are collected in the field. A detailed clinical examination, including glaucoma diagnostic procedures, is conducted at the examination centre. Pedigree ascertainment and genetic studies are performed for subjects with occludable angles or glaucoma. Data are recorded in a computerised database. CONCLUSIONS: This study is expected to result in an estimation of the prevalence and a better understanding of the genetics of glaucoma in this region. PMID- 14566636 TI - Cataract blindness in Paraguay--results of a national survey. AB - PURPOSE: To estimate the burden of visual loss and blindness due to cataract in people aged 50 years and over in Paraguay. METHODS: Forty clusters of 60 persons each who were 50 years and older (2400 eligible persons) were selected by systematic random sampling from the entire population of Paraguay. A total of 2136 persons were examined (89% coverage). RESULTS: For the population 50 years and over, the age- and gender-adjusted prevalence of bilateral blindness (VA < 3/60 with available correction) was 3.14% (95% CI: 2.2-4.4). The adjusted prevalence of bilateral cataract blindness (VA < 3/60) was 2.01% (95% CI: 1.3 3.0), making cataract the major cause of bilateral blindness in this age group (64%). The adjusted prevalence of bilateral severe visual impairment (VA < 6/60 with available correction) was 5.17% (95% CI: 3.9-6.7) and the adjusted prevalence of severe visual impairment due to bilateral cataract (VA < 6/60) was 3.09% (95% CI: 2.2-4.3). The cataract surgical coverage (persons) was 44% for bilaterally blind persons with VA < 3/60; 36% for persons with bilateral VA < 6/60; and 28% for any eye with VA < 6/60 due to cataract. With IOL implantation, 77% of the operated eyes could see 6/18, against 46% of the non-IOLs (p < 0.005), a significant better outcome. CONCLUSION: There is a need to increase the cataract surgical coverage in Paraguay. The number of eye surgeons is adequate but the accessibility of cataract surgical services in rural areas and the affordability of surgery to large sections of society are major constraints. PMID- 14566641 TI - Who should pay for the new medical treatments in uveitis? PMID- 14566642 TI - Value-based medicine: evidence-based medicine and beyond. AB - Value-based medicine is the practice of medicine emphasizing the value received from an intervention. Value is measured by objectively quantifying: 1) the improvement in quality of life and/or 2) the improvement in length of life conferred by an intervention. Evidence-based medicine often measures the improvement gained in length of life, but generally ignores the importance of quality of life improvement or loss. Value-based medicine incorporates the best features of evidence-based medicine and takes evidence-based data to a higher level by incorporating the quality of life perceptions of patients with a disease in concerning the value of an intervention. Inherent in value-based medicine are the costs associated with an intervention. The resources expended for the value gained in value-based medicine is measured with cost-utility analysis in terms of the US dollars/QALY (money spent per quality-adjusted life-year gained). A review of the current status and the likely future of value-based medicine is addressed herein. PMID- 14566643 TI - Host defense against bacterial keratitis. AB - PURPOSE: To define factors that protect the eye from Staphylococcus aureus keratitis and limit tissue damage once keratitis occurs. METHODS: Rabbit tears were analyzed for bactericidal and phospholipase A(2) (PLA(2)) activities on S. aureus. Inhibition by spermidine of PLA(2) anti-staphylococcal activity in tears was tested in vitro and in vivo. Rabbits immunized with heat-inactivated alpha toxin were challenged with intrastromal injection of S. aureus. RESULTS: Arachidonic acid was cleaved from S. aureus by purified PLA( 2) or rabbit tears. Spermidine inhibited these reactions in vitro and facilitated keratitis in vivo. PLA(2) activity decreased with advanced age and shortly following sleep, but increased with keratitis. Antibody to alpha-toxin significantly reduced corneal damage and epithelial cell sloughing during keratitis. CONCLUSIONS: PLA(2) is a major host-defense component of rabbit tears. Alpha-toxin is a major mediator of corneal damage, and antibody to alpha-toxin reduces pathologic changes during keratitis. PMID- 14566645 TI - DNA microarray analysis of the uninoculated eye following anterior chamber inoculation of HSV-1. AB - PURPOSE: To use DNA microarray to analyze the expression patterns of genes in the uninoculated eye following uniocular anterior chamber inoculation of HSV-1. METHODS: On Day 9 following inoculation of 2 x 10( 4) PFU of HSV-1 (KOS strain) or an equivalent volume of tissue culture medium into one anterior chamber of BALB/c mice, the uninoculated eyes were enucleated, pooled, and total RNA was isolated. cDNA was synthesized from the total RNA. The gene expression patterns were inferred based on the hybridization intensities of the probes on the cDNA array. The hybridization signals were globally normalized and filtered. The data were analyzed using hierarchical and gene tree clustering algorithms. Additional uninoculated eyes collected on Day 9 p.i. were stained for F4/80 and CD19. RESULTS: Compared with the uninoculated eye of control mice, 3800 genes were upregulated at least twofold in the contralateral eye of HSV-1-infected mice. Among the 10 most upregulated genes, T cell-specific protein, MHC II antigen A, and MHC II k region locus 2 were upregulated 179-, 164-, and 162-fold, respectively. Ten T-cell receptor-related genes, 61 cytokine and chemokine genes, and 16 MHC genes were upregulated. Furthermore, 11 immunoglobulin and B cell genes and 11 macrophage-related genes were also upregulated. F4/80+ and CD19+ cells were observed on Day 9 p.i. CONCLUSIONS: The DNA microarray results support the idea that T cells and immunomodulatory factors (cytokines, chemokines) are likely to be involved in HSV-1 retinitis. These results also suggest that B cells and/or macrophages play a role in the pathogenesis of HSV-1 retinitis. PMID- 14566646 TI - Systemic and ocular antibody responses to inactivated acute hemorrhagic conjunctivitis (AHC) virus; enterovirus 70 (EV70). AB - PURPOSE: Evaluate the immune response in rabbits injected with EV70, the agent of acute hemorrhagic conjunctivitis (AHC) and AHC associated neuropathy. METHODS: Rabbits were injected intramuscularly with uv-light inactivated EV70 isolate J670/71. Neutralizing activity against EV70 was quantified in serum and tear samples and the immunoglobulin (Ig) classes of the neutralizing activity in serum identified by sucrose gradient ultra-centrifugation. Adjuvant muramyl dipeptide (MDP) was applied topically to assess the role of ocular inflammation on levels of neutralizing antibody, proteins and Ig in tears. The protective effects of human and rabbit sera and interferon-alpha (IFN-alpha) against EV70 were compared in human conjunctival and lens cells. RESULTS: Sera collected at 6 and 13 d contained 19S IgM anti-EV70 neutralizing antibody, while serum collected 21 d post injection contained 19S IgM and 7S IgG anti-EV70 neutralizing antibody. Low titers of anti-EV70 activity (< or =30 U/ml) were detected in tears of seropositive rabbits. MDP induction of conjunctivitis in seropositive rabbits increased tear IgG concentration (3-fold) and anti-EV70 neutralizing antibody titers (> or =10-fold). The protective effect of the rabbit and human sera against EV70 infection in conjunctival, but not lens epithelial cells, was enhanced by the addition of IFN-alpha. CONCLUSIONS: Immunization with uv-light inactivated EV70 elicits a classical humoral immune response in rabbits. The protective activity of serum in EV70-infected human conjunctival cells, but not lens cells, was increased by IFN-alpha. Adjuvant MDP-induced conjunctivitis, increased blood-conjunctival barrier (BCB) permeability and anti-EV70 neutralizing activity in tear of seropositive rabbits. The results suggest immunization with inactivated EV70 could provide systemic as well as ocular protection during natural EV70 infection. PMID- 14566647 TI - Human mast cell subtypes in conjunctiva of patients with atopic keratoconjunctivitis, ocular cicatricial pemphigoid and Stevens-Johnson syndrome. AB - PURPOSE: Investigate mast cell (MC(S)) subtypes in atopic keratoconjunctivitis (AKC), ocular cicatrical pemphigoid (OCP), and Stevens-Johnson Syndrome (SJS). METHODS: MC(S) subtypes were determined by immunohistochemistry of conjunctiva (obtained from 34 patients--9 AKC, 9 OCP, 9 SJS and 7 normal) using monoclonal antibodies directed against chymase (MC(C)) and tryptase (MC(T)). Double staining was used to distinguish MC(S) as positive for both chymase and tryptase (MC(TC)). RESULTS: The number of MC(S) was significantly increased in AKC, OCP and SJS patients, compared to normal subjects. MC( C) were especially high in AKC, and moderately high in OCP. MC(T ) and MC(TC) were similar in each disease group. CONCLUSIONS: While the AKC findings were not surprising, the result in OCP and SJS suggests that MC(S) play an underappreciated role in the inflammatory process of these disorders. Disparate proportions of MC(S) subtypes in these diseases may imply differential functions of MC(S) in these disorders. PMID- 14566648 TI - Tear eotaxin levels in giant papillary conjunctivitis associated with ocular prosthesis. AB - PURPOSE: To examine the role of eotaxin and eosinophil recruitment in the immunopathogenesis of ocular prosthesis-associated giant papillary conjunctivitis (GPC). METHODS: The tear eotaxin level was measured in 68 eyes with GPC, the fellow eyes of the GPC patients, and 22 normal subjects, using an ELISA method. Upper tarsal conjunctival specimens harvested from 18 patients with GPC were examined by light microscopy. RESULTS: There was no significant difference in tear eotaxin levels between patients with GPC and healthy subjects. In patients with chronic GPC, the tear eotaxin levels were significantly lower. These eyes also had decreased conjunctival cellularity and increased fibrosis in the substantia propria. Biopsy specimens showed infiltration of lymphocytes and mast cells, but no eosinophils were found. CONCLUSION: Eotaxin and eotaxin-mediated eosinophil recruitment do not seem to have a major role in the immunopathology of chronic GPC associated with an ocular prosthesis. PMID- 14566649 TI - Optic nerve dysplasia and renal insufficiency in a family with a novel PAX2 mutation, Arg115X: further ophthalmologic delineation of the renal-coloboma syndrome. AB - Renal-coloboma syndrome, an autosomal dominant disorder associated with mutations in PAX2, is characterized by colobomatous eye defects, renal hypoplasia, vesicoureteral reflux, high-frequency hearing loss, and rarely central nervous system abnormalities. We identified a three-generation family with optic nerve colobomatous dysplasia and renal disease. We report the identification of a novel mutation in PAX2 in this family with renal-coloboma syndrome, Arg115X. We also report on the ocular and extraocular manifestations of PAX2 mutations for all cases of renal-coloboma syndrome reported to date. PMID- 14566650 TI - Phenotypic variability in three carriers from a family with choroideremia and a frameshift mutation 1388delCCinsG in the REP-1 gene. AB - PURPOSE: To perform genotype-phenotype correlations in a family with choroideremia. METHODS: A three-generation family with two affected males and five carriers was the subject of the study. Molecular genetic analysis using single-strand conformation polymorphism analysis (SSCP) was conducted in all subjects, while electroretinography (ERG), multifocal ERG (mfERG), scanning laser ophthalmoscope microperimetry (SLO perimetry), fluorescein angiography, and Arden contrast color testing were performed in one male and three carriers. RESULTS: The findings in the affected male were typical for advanced choroideremia. The three carriers demonstrated a variable clinical phenotype including reduction of visual acuity and ERG and angiographic changes in one. Molecular genetic analysis revealed a functional null mutation (1388delCCinsG) in the REP-1 gene. CONCLUSIONS: A severe retinal pathology was found in the affected male, indicating that the 1388delCCinsG is a severe mutation. Varying phenotypes were present in the three carriers examined. The phenotype in carriers has been explained by random X-inactivation with varying expression of the inactivated and activated gene copy inside the same cell of both the retinal pigment epithelium and the rods. This thesis is in agreement with the clinical data obtained here. PMID- 14566652 TI - A rare homozygous rhodopsin splice-site mutation: the issue of when and whether to offer presymptomatic testing. AB - Having identified a disease-associated rhodopsin mutation in a patient with retinitis pigmentosa (RP), the issue is to address the question of whether to offer genetic testing to at-risk family members. Two members of a South African (SA) family, one of whom suffers from RP, as well as 54 unrelated SA RP patients from the same population group were investigated using single-stranded conformational polymorphism analysis followed by DNA sequencing. A rare homozygous mutation at the intron-exon boundary of exon 4 in the rhodopsin gene was identified in the proband. One of his siblings was found to be heterozygous for the same mutation. The mutation was not detected in the 54 unrelated SA RP patients examined, 11 of whom were sporadic cases. A low incidence of RP amongst heterozygous carriers of this mutation has been reported; however, in the past it has been unclear whether the mutation has an effect in single copy or dual copy. To the best of our knowledge, this is the first time that this mutation has been reported as homozygous in an affected individual, thereby resolving the issue and confirming that it is a recessive disease-associated mutation. This is also the first autosomal recessive RP disease-causing rhodopsin mutation that has been identified in Southern Africa. Analysis of the extended pedigree indicated obligate heterozygous carriers of the mutation, without obvious signs of visual impairment in early adulthood. The extent to which potential heterozygous carriers should be pursued and clinically examined is discussed and the question is addressed as to whether to inform the family of these molecular findings. PMID- 14566651 TI - Clinical studies of X-linked retinitis pigmentosa in three Swedish families with newly identified mutations in the RP2 and RPGR-ORF15 genes. AB - PURPOSE: To describe new disease-causing RP2 and RPGR-ORF15 mutations and their corresponding clinical phenotypes in Swedish families with X-linked retinitis pigmentosa (XLRP) and to establish genotype-phenotype correlations by studying the clinical spectrum of disease in families with a known molecular defect. METHODS: Seventeen unrelated families with RP and an apparent X-linked pattern of disease inheritance were identified from the Swedish RP registry and screened for mutations in the RP2 and RPGR (for the RP3 disease) genes. These families had been previously screened for the RPGR exons 1-19, and disease-causing mutations were identified in four of them. In the remaining 13 families, we sequenced the RP2 gene and the newly discovered RPGR-ORF exon. Detailed clinical evaluations were then obtained from individuals in the three families with identified mutations. RESULTS: Mutations in RP2 and RPGR-ORF15 were identified in three of the 13 families. Clinical evaluations of affected males and carrier females demonstrated varying degrees of retinal dysfunction and visual handicap, with early onset and severe disease in the families with mutations in the ORF15 exon of the RPGR gene. CONCLUSIONS: A total of seven mutations in the RP2 and RPGR genes have been discovered so far in Swedish XLRP families. All affected individuals express a severe form of retinal degeneration with visual handicap early in life, although the degree of retinal dysfunction varies both in hemizygous male patients and in heterozygous carrier females. Retinal disease phenotypes in patients with mutations in the RPGR-ORF15 were more severe than in patients with mutations in RP2 or other regions of the RPGR. PMID- 14566653 TI - Electrophysiology and ocular blood flow in a family with dominant optic nerve atrophy and a mutation in the OPA1 gene. AB - OBJECTIVE: To characterize the clinical phenotype, with emphasis on electrophysiology and blood flow measurements, of a family with dominant optic nerve atrophy and an identified mutation in the OPA1 gene. METHODS: Seven family members were examined. Ophthalmological evaluation included testing of visual acuity, ophthalmolscopy, kinetic perimetry, color vision testing, full-field electroretinography (ERG), multifocal electroretinography (MERG), and multifocal visual evoked potential (MVEP). Retrobulbar arterial blood flow and retinal capillary perfusion was measured in three patients using scanning laser Doppler flowmetry (SLDF) and color Doppler imaging techniques. PCR-SSCP and DNA sequencing determined the presence of a mutation in exon 18 of the OPA1 gene. RESULTS: The clinical characteristics varied considerably in the family. The ERG and the MERG demonstrated normal retinal function, while the MVEP was abnormal in all examined patients. Retinal and optic nerve head capillary perfusion was significantly decreased in the three patients examined with SLDF. Retrobulbar blood flow velocities were significantly decreased in the central retinal and ophthalmic arteries. In all seven examined subjects, a microdeletion (1756 1767del(12 bp)) in the OPA1 gene was identified. CONCLUSION: Patients with a mutation in the OPA1 gene have a very variable phenotype. MVEP and blood flow measurements are two new objective methods for an easier detection of this specific genetic optic nerve atrophy. PMID- 14566654 TI - Retinopathy of incontinentia pigmenti: a case report with thirteen years follow up. AB - A spectrum of retinal features has been recognised in association with incontinentia pigmenti. The majority of reported cases describe a rapidly progressive proliferative retinopathy, often leading to retinal detachment, emphasizing the potentially blinding nature of the disease. In some instances, the retinopathy has been noted to be less severe but follow-up has been short. We describe a young girl with incontinentia pigmenti in whom the retinopathy has shown a fluctuating but stable course over 13 years without treatment. PMID- 14566658 TI - [Minimally-invasive therapy of urinary stones]. AB - Open surgery was the standard therapy for urinary calculi up to about 30 years ago. This changed upon introduction of extracorporeal shockwave lithotripsy (ESWL) in 1980, a procedure that is now the primary therapy for 70 % of the patients in western countries. Simultaneously, endourological procedures like ureterorenoscopy (URS) and percutaneous nephrolithotripsy (PCNL) have been improved, and now, modern small diameter and highly efficient instruments offer an ideal alternative to shockwave lithotripsy. Today, minimally-invasive stone treatment has replaced open stone surgery almost completely. This article introduces ESWL, URS and PCNL and discusses indications, outcomes and limitations. PMID- 14566659 TI - [Perioperative immune modulation in renal cell carcinoma--review of the current situation]. AB - Surgery and immunotherapy are the mainstay in the treatment of renal cell carcinoma. Surgery, however, is associated with considerable perioperative immunodysfunction. This immunodysfunction can be modulated by pretreatment with Interleukin-2 (IL-2), which appears to prolong tumour-specific survival. Perioperative immunomodulation could help close the therapeutic gap between neoadjuvant and adjuvant immunotherapy. This article reviews recent literature and presents original data of 63 patients. PMID- 14566660 TI - [Gap junctions in the human urinary bladder]. AB - PURPOSE: Gap junctions are intercellular contacts important for the synchronization of muscle cell activity through electrical coupling. Since the role of gap junctions for the function of smooth bladder muscle is still a matter of debate, we investigated the occurrence of gap junctions and the gap junctional protein connexin (Cx) 45 in the detrusor of the nonobstructed stable human bladder. MATERIALS AND METHODS: Detrusor biopsies from 6 patients aged 64 (55-72) years with stable nonobstructed bladders were investigated for the occurrence of gap junctions by electron microscopy, molecular biological techniques and immunohistochemistry. RESULTS: Transmission electron microscopy and freeze fracture showed the presence of gap junction at plasma membranes of detrusor smooth muscle cells. By reverse transcriptase (RT) polymerase chain reaction (PCR) and in situ hybridization, we found an expression of Cx45 in the detrusor. These data were confirmed by immunolocalization of Cx45 on smooth muscle cells. CONCLUSIONS: This study provides morphological as well as molecular biological and immunohistochemical evidence that bladder smooth muscle cells are electrically coupled. PMID- 14566661 TI - [Transitional cell carcinoma of the prostate in cystoprostatectomy specimens]. AB - PURPOSE: Transitional cell carcinoma (TCC) of the prostate/prostatic urethra is a risk factor for urethral recurrence after radical cystoprostatectomy for TCC. Using conventional sectioning techniques, prostate involvement (prostatic urethra, acini, ducts and/or stroma) has been detected in a range of 10-20% of the patients, whereas transversal whole mount sectioning has revealed 43 % prostate involvement in two reported series. Due to different mechanisms of prostate involvement (intraurethral, extravesical and direct overgrowth into the prostatic stroma), preoperative transurethral biopsies of the prostate might not accurately determine such involvement. In this study we examine the prostate using a longitudinal whole mount sectioning technique, correlate TCC of the prostate with the characteristics of the bladder tumour and, thus, validate the preoperative transurethral resection biopsies. MATERIAL AND METHODS: Patients scheduled for cystoprostatectomy or cystoprostatourethrectomy were investigated by preoperative resection biopsies from the prostatic urethra and mapping of the bladder. The cystectomy specimen was fixated with the bladder filled with formalin, and the prostate and bladder neck examined using longitudinal whole mount sectioning. RESULTS: In 13 of the 43 (30%), patients TCC was identified in the prostate. Of these 13 patients, 9 had been identified in the preoperative resection biopsies from the prostatic urethra. Of the patients with prostatic involvement, 46% had carcinoma in situ (Cis) in the bladder neck/trigone and 38% had multifocal Cis in the bladder. Comparing this to the group of patients without prostatic involvement, the respectively figures are 20% and 23%. A tumour in the trigone, either invasive or Cis, was detected in 5/13 patients with prostatic involvement as compared to one patient (3%) without TCC of the prostate. Multiple bladder tumours were more common in patients with prostatic involvement and were larger (3.2 cm compared to 2.2 cm). CONCLUSIONS: Preoperative resection biopsies from the prostatic urethra do not always detect TCC in the prostate. Cis in the bladder neck/trigone or multifocal and multiple bladder tumours could be risk factors for prostate involvement of TCC. PMID- 14566662 TI - [Quality of life following radical prostatectomy]. AB - PURPOSE: To investigate quality of life in prostate cancer patients after radical retropubic prostatectomy. MATERIALS AND METHODS: 401 men who underwent radical prostatectomy between January 1995 and September 1999 at our clinic were asked for information concerning daily life activities, overall satisfaction, voiding ability and sexual activity in standardized questionnaire. RESULTS: The average follow-up was 36 months, 72 % (n = 289) of our patients replied. In 53 (18%) of the men who replied to our questionnaire, the nerve-sparing technique had been used. At the time our patients were questioned, 202 (70%) did not use any pads, and 211 (73%) were satisfied or very satisfied with the outcome. Of these patients 98% would undergo a radical prostatectomy again. 13% (n = 37) of our patients reported limited satisfaction, 14% (n = 41) were not satisfied with their postoperative status. However, 69% of this latter group would nevertheless decide to have a radical prostatectomy again. Of the patients who underwent the nerve-sparing technique, 68% (n = 36) reported sufficient erections for sexual intercourse, four of them under medication with sildenafil. Patients who underwent the nerve-sparing technique reported better results in daily life activities, general health status, IPSS-score and continence. While age at the time of radical prostatectomy did not influence life quality, we found substantial correlation with the postoperative tumor stage. CONCLUSIONS: Our results underline the importance of early prostate cancer detection, not only regarding a lower recurrence rate, but also because of a better postoperative quality of life. PMID- 14566663 TI - [The effect of enterocystoplasty in childhood on linear growth]. AB - OBJECTIVE: The assumption that enterocystoplasty in children has a detrimental effect on linear growth has been based almost exclusively upon a chance finding in a retrospective study 10 years ago. We re-evaluated the same research question in a larger cohort and with a longer follow-up. PATIENTS AND METHODS: Between 1982 and 1997, 242 children and adolescents underwent enterocystoplasty. Patients with conditions involving organ systems apart from the urinary tract, and those with myelomeningocele, malignant diseases, reduced glomerular filtration rate and incomplete notes were excluded. In the definitive study cohort (123; mean age at operation 8.6 years; mean age at investigation 16.8 years), enterocystoplasty had been undertaken using colon in 70, ileum in 37, a combination of both in 11, ileocaecal segments in three and stomach in two patients. RESULTS: In all, 1215 height and weight measurements had been recorded. The distribution of percentile positions before and after enterocystoplasty showed a normal configuration, with 83 % and 80 % of patients growing within two standard deviations of the 50th percentile. After surgery, 85 % either remained the same or reached a higher percentile. Nineteen (15.5 %) were in a lower position, with a similar tendency in the weight percentile. A clinically relevant growth disorder was recognized in four patients with a complete endocrinological evaluation; in none of these was enterocystoplasty thought to be a causal factor. CONCLUSIONS: It is very unlikely that loss of the preoperative percentile position on the growth curve in 15 % of children after enterocystoplasty is a consequence of that particular surgery. Rather, it is a non-specific phenomenon that has to be considered in any clinical population of the same size and age distribution after the same length of time. PMID- 14566665 TI - [Sarcoidosis of the spermatic cord and epididymus]. AB - INTRODUCTION: Sarcoidosis (Boeck's disease) is of unknown etiology with an incidence of 10 - 40/10(5) people. In 75% of the cases, pulmonary manifestations are found, with histological findings of noncaseating epithelial granulomas. Bilateral enlargement of the hilus lymph nodes are common and divided into three grades of enlargement. The initial symptoms are cough, fever and dyspnea. CASE REPORT: We report a man, 43 years old, first admitted to our hospital in 1988 with a tumor at the right spermatic-cord. Histological examination revealed a granulomatous inflammation, fitting to sarcoidosis, which was verified in the same year by mediastinoscopy. Partial resection of the left epididymal testis was performed in 2001 due to marked rigidity and histological fibrosis. CONCLUSIONS: Urological manifestation of Boeck's disease is rare. Approximately 30 cases have been described in literature with various urological associations. In most cases, urological sarcoidosis is found accidentally during examination of the most common form, pulmonary sarcoidosis. PMID- 14566664 TI - [Rupture of a renal artery aneurysm during delivery]. AB - INTRODUCTION: Renal artery aneurysms represent a rare entity with an incidence of 0.3 - 1.3% in angiographies. Rupture of such aneurysms are rare. During pregnancy there is an increased risk of rupture with a high mortality for mother and child. We report the first case of rupture of a left renal artery aneurysm during delivery with survival of mother and child. CASE REPORT: Due to arrest of labor during a home birth, the patient was admitted to the hospital, where an immediate forceps extraction on account of fetal asphyxia was performed. Immediately after delivery a hypovolemic shock developed. Ultrasound examination showed a huge subsplenic hematoma. Emergency laparotomy revealed a ruptured renal artery aneurysm. The renal artery was ligated on the assumption of a lower pole vessel. Despite hemodynamic stability a drop in hemoglobin occurred the following day. Angiography showed no perfusion of the left kidney. A left nephrectomy was performed without complications. The further course was uneventful. Mother and child survived. CONCLUSIONS: Rupture of aneurysms of the splenic and renal artery during later stages of pregnancy, during delivery or after childbirth are possible etiologies in cases of unexplained shock and abdominal pain. An emergency angiography in such cases helps to reveal the underlying cause and renders possible a targeted operative approach. PMID- 14566667 TI - Extensive surgery in metastatic testicular cancer. AB - Surgery remains an important component in the multimodal treatment of patients with advanced testicular cancer. Recently, however, the indications for post chemotherapy residual tumor resection have changed. Patients with advanced seminoma very rarely need surgical resection of the residual disease after standard chemotherapy. In contrast, patients with high stage non-seminomatous testis cancer must undergo post-chemotherapy surgery in most cases. Surgical resection in metastatic disease may also be necessary in patients with recurrent tumors, patients with persisting marker elevation during chemotherapy and patients with late relapses. Post-chemotherapy residual tumor resections, "redo" retroperitoneal tumor operations and other salvage resections are often technically demanding procedures with unusual surgical approaches that require individualized perioperative planning in order to reduce morbidity. This paper summarizes the current indications for post-chemotherapy surgery and discusses various surgical approaches and techniques, perioperative management recommendations, as well as complications of these extensive resection procedures. PMID- 14566668 TI - Perineal complications following radical perineal prostatectomy. AB - PURPOSE: Radical perineal prostatectomy has recently attracted renewed interest and wider acceptance as an alternative route to the retropubic approach. While presumed lower morbidity is one reason for perineal prostatectomy we evaluated our patients for complications that are specific for the perineal approach. PATIENTS AND METHODS: We have retrospectively analyzed 412 patients who underwent perineal prostatectomy from 10/1996 to 12/2000. Patients for the perineal approach were selected on the base of preoperative PSA (10 ng/ml) and biopsy Gleason score (< 7) without the need for simultaneous lymphadenectomy. A cystogram was performed routinely on day 7 p. o. and the catheter removed when patent anastomosis was confirmed. Intra-, peri- and postoperative complications were recorded and evaluated. RESULTS: Intraoperative complications encompassed 22 cases of rectal injury (5.5 %) with standard primary 2-layer closure, however 4 patients subsequently developed a stool fistula and 3 required a colostomy. Hematoma in the prostatic fossa was diagnosed in 21 patients (5.2 %) and was removed surgically due to infection or increasing size in 4 patients. In 6.5 % of the patients acute urinary retention occurred after catheter removal (91 % after 7 days), while 4.2 % showed urinary extravasation via the perineal wound. Both incidents healed uneventfully with prolonged catheterization in all except 2 cases with concomitant hematoma in whom open fistula closure with a tunica vaginalis graft was performed. The rate of perioperative transfusion was 6.4 %, transient paresthesia and weakness of the leg were observed in 3 patients. CONCLUSIONS: Radical perineal prostatectomy seems to be a procedure with a low rate of complications and surgical reinterventions. However, the spectrum of observed complications is different to that of retropubic prostatectomy and requires specific management. PMID- 14566669 TI - Simplified orthotopic ileocecal pouch (Mainz pouch) for bladder substitution. AB - OBJECTIVE: To report operative modifications that render construction of the orthotopic Mainz pouch more simple, namely formation of the pouch using absorbable staples and utilization of the ileocecal valve as an antireflux mechanism. PATIENTS AND METHODS: Between January 2001 and May 2002 out of 92 cystectomy cases, 21 patients (20 males and 1 female) underwent a simplified orthotopic Mainz pouch. The ureters were implanted via Wallace or Nesbit technique in the supravalvular portion of the ileocecal valve. The mean (range) age was 58 (43 - 74) years and the mean follow-up is 11.6 months (range 1 - 27). RESULTS: No staple-related complications were encountered. Four renal units (8.8 %) were dilated, one secondary to an impacted ureteral stone and three secondary to ureteral stenosis requiring reimplantation. At 6 months follow-up, 5 renal units showed reflux (11.1 %) but without any clinical symptoms. 86.5 % of patients void spontaneously without significant residual urine. Continence was achieved in 74 % of cases during day and in 74 % at night. CONCLUSION: The simplified Mainz pouch is a versatile form of continent urinary diversion. The use of absorbable staples has simplified the pouch creation and yields functional results which favorably compare with other pouch forms. The ileocecal valve serves as an anti-reflux-mechanism. PMID- 14566670 TI - Laparoscopic radical retropubic prostatectomy combined with approaches through a small open incision. AB - PURPOSE: The laparoscopic technique is now applied to radical prostatectomy. However, even in laparoscopic prostatectomy, we need a small open wound to remove the prostate from the abdomen. We have developed a modified technique of extraperitoneal laparoscopic radical prostatectomy, exploiting this small open wound as a route for surgical manipulations as well. Here, we described our technique and its initial outcomes. PATIENTS AND METHODS: An extraperitoneal retropubic space was developed with finger manipulations through a 3- to 5-cm long suprapubic incision. Three or four trocar ports were set up. A specially designed abdominal wall-lifter was applied to create an endoscopic working space. The surgeons worked using open or endoscopic manipulations through the suprapubic incision or the trocar ports. From October 2000 to August 2001, 11 patients with prostate cancer underwent this surgery. RESULTS: We completed surgery endoscopically without major complications except in one case in which we could not identify a bleeding source. Surgical time ranged from 229 to 469 min. Blood loss ranged from 550 to 3797 ml including urine spilled in the surgical field. Urinary continence returned in 10 cases at 1 to 8 months after surgery. One patient still needed pads at 3 months after surgery. CONCLUSION: Our technique allowed us to avoid insufflation of the abdomen with gas and intraperitoneal surgical intervention that are disadvantages of conventional laparoscopic prostatectomy, offering the same advantages as conventional laparoscopic prostatectomy. PMID- 14566671 TI - Significance of early biopsy in pediatric kidney transplantation. AB - Long-term renal allograft survival is limited mainly by the progressive process termed chronic allograft nephropathy (CAN) or chronic rejection. A pathological feature of CAN is characterized by progressive interstitial fibrosis. Transforming growth factor (TGF)-beta(1) plays an important role in fibrogenesis. We investigated whether the degree of TGF-beta(1) expression in early biopsy specimens routinely obtained from stable allografts at 100 days could predict fibrosis and graft dysfunction in the late phase by immunohistochemistry. Patients were children with a graft from related donors. We immunohistochemically determined intracellular and extracellular expression of TGF-beta(1) in the graft at 100 days using LC antibody (LC) for intracellular TGF-beta(1) and CC antibody (CC) for extracellular TGF-beta(1). We used the change in creatinine clearance between 100 days and 3 years after transplantation (Delta Ccr) as an index of long-term graft function. Image analysis was used to calculate the relative area involved by interstitial fibrosis in trichrome-stained sections of graft biopsy specimens at 100 days and 3 years, designating the change as Delta FI. Delta Ccr was - 4.2 +/- 9.4 mL/min in subjects with minimal early immunoreactivity for CC and - 20.5 +/- 5.9 mL/min in subjects with strong reactivity (p < 0.05). Delta Ccr was - 14.5 +/- 18.6 mL/min in subjects with minimal early immunoreactivity for LC and - 11.7 +/- 12.8 mL/min in those with strong reactivity. Delta FI in subjects with minimal CC reactivity (1.28 +/- 4.11 %) tended to be lower than in subjects with strong reactivity (8.45 +/- 15.47 %). Neither fibrosis at 100 days nor Delta FI differed between subjects with minimal and strong LC reactivity. Thus, extracellular TGF-beta(1) expression in grafts at 100 days after transplantation has an influence on long-term graft function and tends to be associated with increased graft fibrosis at 3 years. PMID- 14566672 TI - Rectosigmoidal bladder utilizing an intussuscepted ileal segment: a surgical technique for urinary diversion. AB - A novel technique of urinary diversion was designed by interposition of an intussuscepted ileal segment between the ureters and the rectosigmoidal pouch, thus preventing ureteral reflux as well as stenosis at the uretero-enteric anastomosis, possible occurrence of urocolonic carcinoma, and frequent evacuation or incontinence. Since all the procedures were limited to the lower abdominal cavity, the surgical invasiveness was compatible with that of an ileal conduit. Our early experience in 15 patients showed that this technique can be considered for those in whom the urethra is not available. PMID- 14566674 TI - Laparoscopic radical nephrectomy for renal cell carcinoma. AB - PURPOSE: To estimate the efficacy of the laparoscopic radical nephrectomy we analyzed the clinical data of our series. PATIENTS AND METHODS: One hundred eighty five patients were enrolled in our laparoscopic radical nephrectomy program between July, 1992 and July, 2001. Of the 185 patients, 146 had small renal tumors (smaller than 5 cm in diameter) and 39 had large tumors (equal to or more than, 5 cm in diameter). Under a laparoscope the kidney, adrenal gland, and perirenal fatty tissue were dissected in an en bloc fashion. In case of taking out a small tumor, the specimen was fractionated within the sack to avoid an additional skin incision after entrapping in the laparoscopy sack in the working space. In case of a large tumor, regional lymph nodes dissection was done and the specimen was taken out intact in the sack through an enlarged incision. RESULTS: Our laparoscopic procedure was successful in 171 of the 185 cases; 14 patients required open surgery because of bleeding from an injured vessels or treatment for other injured organs. The mean operative time was 4.7 hours for both small and large tumors. Estimated blood loss was between 237 and 380 ml on average for small and large tumors, respectively. Full convalescence was achieved around 3 weeks after operation in both groups. Only one patient who had large tumor was found to have micrometastasis in 1 of 5 regional lymph nodes. Recurrences were observed in 4 cases of the small tumor group and in 2 cases of the large tumor group during 1 to 108 months of follow-up. CONCLUSION: Laparoscopic radical nephrectomy is a very useful and safe surgical procedure for renal cell carcinoma. PMID- 14566673 TI - First year of mass screening for prostatic cancer in Yaizu city. AB - Three thousand one hundred and twenty men out of the target population of 22,195 men underwent PSA (prostatic specific antigen) determinations as the mass screening test for CAP (prostatic cancer). Three hundred and sixty men showed high (> or = 4.0 ng/ml) PSA values and proceeded to the secondary examinations. Prostatic biopsies were done on two hundred and twelve men, 35 % (74) of them had CAP. The detection rate was 2.4 % of the total examinees. PMID- 14566675 TI - The role of chromosome 8p22 deletion for predicting disease progression and pathological staging in prostate cancer. AB - The natural course of human prostate cancer is highly variable, and we still lack reliable tools to predict the patient's outcome. Recent publications suggest that the deletion of chromosome 8p22 has an important role for tumor progression in prostate cancer. Totally, 97 patients (41 Japanese and 56 Swedish) were studied to detect the status of chromosome 8p22 deletion by the fluorescence in situ hybridization (FISH) technique. Seventy-seven underwent surgery (59 radical prostatectomies or 18 lymph node dissections), and the specimens were prepared by touch biopsy. Fine-needle aspiration biopsies (FNAB) were obtained from another non-operative 20 cases. Disease progression was evaluated in 57 patients with a median follow-up of 59 months. 8p22 deletions were detected in 58 (60 %) of all cases. The frequency of 8p22 deletion did not significantly differ between different preparations of specimens (touch biopsy vs. FNAB) as well as between different races (Japanese vs. Swedish). Cases with more than pT3 tumors had a significantly higher frequency of 8p22 deletion than those with pT2 (p < 0.01). Multivariate analysis demonstrated that 8p22 deletion was the strongest parameter to predict disease progression (hazard ratio = 5.75; p = 0.0001). Studies on chromosomal deletions of 8p22 by the FISH technique may serve as a universal genetic marker to optimize the treatment strategy in patients with prostate cancer. PMID- 14566676 TI - Taxol resistance and its reversal by synthetic isoprenoids in human bladder cancer cell line. AB - PURPOSE: We investigated the mechanism of action of reversal agents for taxol resistance in bladder cancers. MATERIALS AND METHODS: We isolated a taxol resistant cell line (KK47/TX30) from a human KK47 bladder cancer cell line (KK47/WT). We characterized KK47/TX30 cells and screened reversal agents for taxol-resistance. RESULTS: KK47/TX30 cells exhibited approximately 700-fold resistance to taxol and cross-resistance to Vinca alkaloids and topoisomerase II inhibitors. Western blot analysis demonstrated P-glycoprotein (P-gp) overexpression in taxol-resistant cells. Drug accumulation and efflux studies showed that the decreased taxol accumulation in the resistant cell line was due to enhanced taxol efflux. We synthesized 31 isoprenoids based on the structure of N-solanesyl-N,N'-bis(3,4-dimethoxybenzyl)ethylenediamine (SDB), which could completely reverse multidrug resistance (MDR) as shown previously. Among those examined, trans-N,N'-bis(3,4-dimethoxybenzyl)-N-solanesyl-1,2-diaminocyclohexane (N-5228) could completely reverse taxol-resistance in KK47/TX30 cells. Results of a structure-activity relationship study of isoprenoids suggested that the following structural features were important for overcoming taxol-resistance: (1) a basic structure of 8 to 10 isoprene units, (2) a cyclohexane ring or benzene ring within the framework, (3) two cationic sites in close proximity to each other, and (4) a benzyl group with 3,4-dimethoxy functionalities with moderate electron-donation. CONCLUSIONS: Taxol-resistance was primarily mediated by P-gp overexpression in KK47/TX30 cells. One of the synthetic isoprenoids, N-5228 could completely reverse taxol-resistance in KK47/TX30 cells. PMID- 14566677 TI - A new method using a bacterium for dissolution of urinary stones. AB - PURPOSE: A bacterium, Thiobacillus thiooxidans, oxidizes inorganic sulfur compounds to sulfuric acid to obtain energy for growth. We examined dissolution of urinary stones by the bacteria and studied optimal conditions for the dissolution in human urine. METHODS: Thiobacillus thiooxidans (IFO No. 13 701) was purchased from Hakko Kenkyujyo, Co., Osaka, Japan. The bacteria were grown stationary for 7 days at 30 degrees C in medium. The pH of the medium was changed from pH 4.82 to 1.95 for 28 days. Growth of the bacteria was also examined in human urine by addition of different amounts of sodium thiosulfite. The dissolution of human urinary stones was examined in the bladder of Wistar rat containing the bacteria (1 x 10(8)) for 7 days. 20 ml of 5 % sodium thiosulfite were injected intraperitoneally once a day. Histological changes of urinary tracts and blood test were also examined. As a control, two rats were examined in the same methods without the bacteria. RESULTS: The number of bacteria was increased in human urine depending on the concentration of sodium thiosulfite. Among different kinds of urinary stones, calcium phosphate stone was most dissolved, 78 % reduction of dry weight in the rat bladder. Calcium oxalate, uric acid and magnesium phosphate stones were not well dissolved. But the stone weights in the control rats were increased twice. Slight inflammation was found in the rat bladder. There were no abnormal findings in the blood test. CONCLUSIONS: Thiobacillus thiooxidans can be used for dissolution of urinary stones. This new technique may be useful for dissolution of fragments after ESWL and crystals covered by a urinary stent. Optimal conditions and safety methods should be developed for clinical use. PMID- 14566678 TI - Expression of cyclooxygenase-2 in primary superficial bladder cancer tissue may predict risk of its recurrence after complete transurethral resection. AB - A new modality is necessary to prevent recurrence of superficial bladder cancer after complete transurethral resection (TUR) because of the high recurrence rate even with current prophylaxis protocols. Prostaglandins (PGs) are known to be produced more in transitional cell carcinoma, and etiologically bladder cancer risk is negatively associated with the intake of non-steroidal anti-inflammatory drugs (NSAIDs), which inhibit cyclooxygenase (COX), the rate-limiting enzyme of the PG production. We have shown the chemopreventive effect of piroxicam, an NSAID, on the N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN)-induced rat bladder cancer model. To avoid gastrointestinal side effects of regular NSAIDs, we also showed the chemopreventive effect of nimesulide, a selective inhibitor of the second isoform of COX, COX-2, which does not affect COX-l house-keeping activity in gastrointestinal mucosa on the same model. We also observed induction of COX-2 protein in the rat bladder tumor. In this study, we screened COX-2 protein expression in primary superficial bladder cancer tissues, to elucidate if COX-2 selective inhibitors can be a candidate chemopreventive agent for bladder cancer recurrence. Five and 6 samples of superficial bladder cancer cases with and without recurrence after complete TUR were examined by immunohistochemical analysis. We found more COX-2 protein positive samples in the cases with recurrence than in cases without recurrence. Even though the number of cases examined is small, this result supports our hypothesis that COX-2 contributes to superficial bladder cancer recurrence, thus, selective COX-2 inhibitors can be a candidate chemopreventive agent for the recurrence. PMID- 14566680 TI - Quantitative analysis of testicular interstitial fibrosis after vasectomy in humans. AB - PURPOSE: Germ cell differentiation, DNA synthesis, and apoptosis can be evaluated quantitatively. On the other hand, an interstitial lesion is difficult to examine. We have focused on the quantitative analysis of testicular interstitial fibrosis after vasectomy. MATERIALS AND METHODS: Forty testicular biopsy specimens from twenty consecutive men were obtained at vasovasostomy. Johnsen's mean score was calculated from testicular biopsy specimens. Percent of interstitial fibrosis was determined quantitatively by the NIH-Image after Masson trichrome staining. RESULTS: A significant increase in interstitial fibrosis was observed along with the obstructive interval (p < 0.001). Johnsen's mean score count did not associate with the obstructive interval. CONCLUSION: Interstitial lesions of testicular physiology and pathophysiology can be evaluated using the NIH-Image. Interstitial fibrosis, but not the intraseminiferous status, reflects the irreversible damage of vasectomized testes. PMID- 14566679 TI - Total cystoprostatectomy in the treatment of locally advanced prostate carcinoma. AB - OBJECTIVE: Locally advanced prostate carcinoma frequently causes lower urinary tract symptoms and is a clinical challenge when radiation and/or hormone therapy fail. We investigated whether cystoprostatectomy with urinary diversion benefits patients with locally advanced prostate carcinoma in terms of quality of life and prognostic outcome. PATIENTS AND METHODS: Between 1989 and 2001, we performed 15 cystoprostatectomies for stage C-D1 prostate carcinoma with bladder neck involvement. Of these patients, 5 underwent ileal conduit, 8 rectal bladder, 1 Koch pouch, and 1 ureterocutaneostomy. All the patients received neoadjuvant and/or adjuvant hormonal therapy. In the same period, 28 patients underwent retropubic prostatectomies and 15 patients received hormone therapy alone for stage C-Dl disease. These patients were included as references. RESULTS: Lower urinary tract symptoms caused by bladder involvement were controlled well until the end of follow-up for all the patients in the cystoprostatectomy group. There was no statistically significant difference in QOL score assessed with the EORTC QLQ-C30 questionnaire between the prostatectomy group and the cystoprostatectomy group, while that in the hormone therapy group was lower than those in the surgery groups. There was no statistically significant difference in 5-year PSA relapse-free survival among cystoprostatectomy, prostatectomy, and hormone therapy groups. Patients in the hormone therapy group died earlier than those in the prostatectomy group (p = 0.02), while those in the cystoprostatectomy group did not. CONCLUSION: These results suggest that total cystoprostatectomy with urinary diversion is a valid option, in terms of disease control and QOL, for prostate cancer patients whose tumor is infiltrating into the bladder. PMID- 14566681 TI - Clinical study of urine NMP 22 (nuclear matrix protein 22) as a tumor marker in urinary epithelial cancer. AB - The prognosis of urinary epithelial cancer is still poor, and early detection of this cancer is strongly desirable. The sensitivity of conventional urinary cytology is not satisfactory enough. It is hoped that a specific tumor marker will be established. In recent years, it has been reported that urine NMP 22 is very useful and that urine BFP is also relatively useful. We have now determined urine NMP22 and BFP and studied their clinical usefulness as a tumor marker. Using patients diagnosed with histologically confirmed urinary epithelial cancer as the subjects, we retrospectively studied the usefulness of NMP 22, BFP and cytology mainly with regard to the sensitivity (positivity rate), and also in relation to atypia, degree of infiltration and clinical course. PMID- 14566682 TI - Hand-assisted laparoscopic nephrectomy. AB - PURPOSE: We report our experience in performing a hand-assisted laparoscopic radical nephrectomy (HALRN) for renal cell carcinoma (RCC) and a hand-assisted laparoscopic live donor nephrectomy (HALDN) for renal transplantation. PATIENTS AND METHODS: The clinical data on 21 patients with HALRN for stage T1N0M0 RCC and 5 living kidney donors with HALDN for renal transplantation were reviewed. We compared the results of 21 HALRNs with those of 15 conventional open radical nephrectomies for stage T1N0M0 RCC. RESULTS: The HALRN for RCC was successfully performed without any major complications in all 21 patients. The mean operation time for the HALRN group was significantly longer than that for the open surgery group (262 minutes versus 219 minutes). However, the mean estimated blood loss in the HALRN group was less than that in the open surgery group (250 ml versus 388 ml), although there was no statistically significant difference. The frequency in usage of analgesia postoperatively in the HALRN group was significantly less than that in the open surgery group (2.5 times versus 7.3 times). The HALDN was also successfully performed without any major complications in all 5 cases. The mean operation time, blood loss and warm ischemic time were 248 minutes, 322 ml and 9.8 minutes, respectively. All donors resumed oral intake of food within 48 hours after the surgery and returned to normal, non-strenuous activity by postoperative day 8. CONCLUSIONS: Our results indicate that a hand-assisted laparoscopic nephrectomy is an effective and safe surgical procedure, and it is less invasive than an open nephrectomy. PMID- 14566683 TI - Detection of circulating MN/CA9 positive renal cell carcinoma cells during operation. AB - Our previous study demonstrated the clear detection of MN/CA9 mRNA in peripheral blood samples from RCC patients. However, approximately 30 % of control blood samples from healthy volunteers showed MN/CA9 expression. We have developed a new primer set and optimized the PCR conditions, now resulting in a specificity of 100 %. In tissue samples, all clear cell type carcinomas but none of the spindle cell type and pleomorphic cell type tumors expressed the MN/CA9 message. Analysis of MN/CA9 messages in peripheral blood samples from RCC patients gave positive results for 0/2, 1/9, 0/4 and 4/12 of stage I, II, III and IV cases, respectively. RT-PCR analysis using preoperative renal venous blood samples resulted in clear detection of MN/CA9 positive cells in 2/4, 3/13, 2/6 and 1/1 of stage I, II, III and IV cases, respectively. Our results suggest that assessment of MN/CA9 expression by RT-PCR is a promising method for detecting cancer cells in the circulation of patients with RCC. PMID- 14566684 TI - An artificial testis for production of rat haploid cells. AB - PURPOSE: We attempted to apply the microgravity cell culture system for rat testicular germ cell maturation in vitro. METHODS: Primary spermatocytes were isolated from immature male rat by sedimentation velocity. Sertoli cells were isolated from another immature male by enzyme digestions. Sertoli cell aggregates were plated into conventional tissue culture flasks and incubated at 37 degrees C for 48 hours. These pretreated Sertoli-enriched monocultures were used in preparing Sertoli cell-primary spermatocyte cocultures. And then, primary spermatocytes and Sertoli cells were cocultured in a microgravity cell culture device for 28 days. RESULTS: Cell viability rate is more than 50 % after a 28-day long period of incubation. Furthermore, about 23 % haploid germ cells are observed. CONCLUSIONS: These results using primary spermatocyte coculture with Sertoli cell aggregates under microgravity show that it is possible to mature these cells up to the round spermatid and even to elongating/elongated steps. It may be possible to overcome the male sterility due to maturation arrest at the primary spermatocyte stage. PMID- 14566685 TI - Experimental and clinical study of a holmium: YAG laser with adjustable pulse duration. AB - BACKGROUND: The holmium:YAG laser is used for treatment of urolithiasis, transurethral laser ablation for benign prostate hypertrophy and bladder tumor. The pulse duration was fixed in the previous Ho:YAG laser systems. We have evaluated more practical pulse durations to disintegrate the stone. The SPHINX Ho40 (Heraeus Corporation) can change the pulse duration freely in the range from 150 microsec. to 800 microsec. MATERIALS AND METHODS: 1) We measured the total energy to perforate through stone models at three different pulse durations (150, 300, 600 microsec.). 2) We experimented with the energy of each single pulse and the power of the shock wave. 3) We observed thermogenesis during the lithotripsy for each pulse duration. 4) Disintegration effects of Ho:YAG are compared with other lithotripsy systems in clinical cases. RESULTS: 1) It was possible that the smallest amount of the total energy went through the fragment at the pulse duration 150 microsec. 2) The maximum amount of energy of the wave is higher when the pulse duration is short. Although the amount of energy was 12.6 V, the amounts of energy at 800 microsec decreased with a pulse duration 150 microsec. to 6.46 V which was about 40 % 3) The highest temperatures achieved when the irradiation of laser was started and finished were compared. The shorter the pulse duration was, the higher was the peak power. The shock wave was also more effective to disintegrate stones on using short pulse durations. 4) The individual clinical success rates are Ho:YAG (85.1 %), Alexandrite (80.6 %) and Lithoclast (74.5 %). PMID- 14566686 TI - [Sacral neuromodulation in patients with nonobstructive, chronic urinary retention: relevance of the carbachol test and influence of associated nerve lession]. AB - OBJECTIVE: Aim of this study was to evaluate whether the carbachol test and the nerve lesions responsible for bladder dysfunction possess predictive value in patients with chronic urinary retention who undergo temporary sacral neuromodulation (PNE test). MATERIAL AND METHODS: In 1999 and 2000, PNE tests were performed in 24 patients with chronic urinary retention and acontractile detrusor previously assessed by urodynamics. In 18 patients, a carbachol test was performed during urodynamics. The diagnosis related to the acontractile detrusor was additionally assessed and compared to the successful outcome of the PNE test. RESULTS: The PNE test was successful in 8 of the 24 patients (33.3 %). The bladder was completely emptied during the PNE test in 3 of the 10 patients with negative carbachol test and in 3 of the 8 patients with positive carbachol test. The highest success rate (80 %) was observed in patients after hysterectomy, whereas after lumbosacral pulposal prolapse or with a CNS tumor, it was only 20 - 33.3 %. CONCLUSIONS: We conclude that sacral neuromodulation is an effective treatment option in patients with nonobstructive urinary retention. The carbachol test does not possess any definitive predictive value with respect to the success rate of sacral neuromodulation in patients with chronic urinary retention. The success rate more likely depends on the localisation of the nerve lesion. PNE tests should be performed in all patients with therapy resistent nonobstructive urinary retention, as no other predictive factors exist. PMID- 14566687 TI - [Clinical Results of PNE Tests in 70 Patients with Different Bladder Dysfunctions]. AB - PURPOSE: Prior to implantation of a sacral neuromodulator we carried out PNE tests (peripheral nerve evaluation test) with bilateral test stimulation to establish which patients might profit from this kind of therapy. In contrast to the original unilateral technique used by Tanagho and Schmidt, we performed bilateral PNE test stimulation. Moreover, we analysed the diagnostic characteristics of those patients who had positive PNE test results and could thus receive a chronic sacral neuromodulator. MATERIALS AND METHODS: We performed bilateral PNE test stimulation in 70 patients (mean age: 53.6 yrs; 41 with retention symptoms and 29 with a hyperactive detrusor) over a minimum of 3 days. Retrospectively, we analysed the distribution of diagnostic characteristics (retention vs. overactive bladder and neurogenic vs. idiopathic) in the implant recipients. RESULTS: All patients received bilateral PNE test stimulation, during which the stimulation amplitudes were adjusted individually for each side. 8 patients were treated with the original PNE-electrode (model 0 041 830 - 002, Medtronic Inc., USA) without success. Of the remaining 62 patients, who were treated with an improved electrode, the PNE test was successful in 32 cases (51.6 %). Twenty-seven of these patients suffered from a neurogenic bladder dysfunction, and, in 5 cases, the causes were idiopathic. CONCLUSIONS: Bilateral PNE-test stimulation and the use of advanced PNE electrodes (model 3057, Medtronic Inc., USA) led to a positive PNE result in 51.6 % of the tested patients. Of these, the group with neurogenic bladder dysfunctions showed the highest response rate. Compared with the success rates in the multicenter study, we were able to increase the overall PNE response rate significantly. For this reason, we prefer a bilateral PNE-test with side-specific stimulation. PMID- 14566688 TI - [Does a Combined Fascial Sling - Burch Colposuspension Display Advantages over a Fascial Sling alone for Treatment of Urinary Stress Incontinence in Females?]. AB - PURPOSE: Despite the development of new surgical techniques, the fascial sling procedure remains an important surgical technique for the treatment of female urinary stress incontinence. An advantage of combining it with an additional Burch colposuspension has been suggested. The objective of our study was to evaluate retrospectively selected patients who had undergone a fascial sling procedure with and without Burch colposuspension. MATERIALS AND METHODS: Of a total of 390 females who underwent an incontinence operation at our department between 1990 and 1999, 56 patients had had a fascial sling plasty. A total of 50 patients (89 %) were followed for a median of 59.5 months. The median age was 60 years. 56 % of the patients displayed recurrent stress incontinence. The previous operations had been performed via a vaginal approach in 42.9 % and an abdominal approach in 57.1 %. The sling procedure used was that of Narik and Palmrich. Of the 50 patients, 14 had an additional Burch colposuspension. RESULTS: The continence rates (no pads) were for patients with a fascial sling procedure alone 63.9 % and for the combination of both operations 64.4 %. An improvement (1-3 pads) was seen in 27.8 % and 21.4 %, respectively. No changes were seen in 5.6 % and 7.1 % and impairment was seen in 2.7 % and 7.1 %, respectively. After a five year follow-up, the total patient satisfaction rate was 78 %. CONCLUSIONS: The fascial sling is effective operative technique for treating female urinary stress incontinence, especially in severe and type III incontinence and in patients who had undergone previous operations for incontinence. The operation is safe and is the only technique that offers controlled overcorrection in desperate cases. An advantage of adding a Burch colposuspension to the fascial sling procedure was not detected in our patient group. PMID- 14566689 TI - [A New Ex-Vivo 3D Bladder Matrix Model for Standardised Evaluation of Urothelial Transfection Methods]. AB - PURPOSE: Gene therapy strategies are a promising new alternative option in the treatment of cancer diseases and great effort is dedicated to the development of new gene transfer methods. At present, in vitro cell culture experiments or in vivo animal trials are the only available alternatives in the search for new gene transfer methods. We attempted to develop and evaluate a new 3D matrix model as a step between in vitro experiments and animal trials. MATERIALS AND METHODS: We generated a convenient model with an agarose gel as a basis for small intestinal submucosa or a polyethylene membrane. The produced model consisting of human smooth muscle cells and human bladder carcinoma cells was transfected with a modified standard Lipofectamine trade mark 2000 transfection procedure and visualised by fluorescence microscopy after cryo-sectioning. RESULTS: With the help of this new technique it is possible to generate three dimensional tissues consisting of different types of cells in which the cells are adherent on the polyethylene and the SIS-membrane during the entire treatment. The resulting model was successfully transfected with the pEGFP-N1 plasmid. CONCLUSIONS: This new three dimensional model allows the standardised evaluation of new transfection methods on multilayered ex-vivo generated tissues consisting of different cells. PMID- 14566690 TI - [Blunt testicular injury - conservative or surgical treatment?]. AB - INTRODUCTION: Testicular trauma can be classified into blunt or penetrating injury. While every case of penetrating trauma is subjected to surgical treatment, the decision between surgery and conservative treatment must be made for each case of blunt injury. Inspection, palpation and duplex sonography may be helpful in finding the correct decision. Herein we present a case report and review of the current literature. CASE REPORT: A 14-year-old boy was admitted to an urological department after blunt scrotal trauma. Ultrasound examination revealed a peritesticular hematoma. After 9 days of conservative treatment, surgical exploration and orchidectomy for rupture of the tunica albuginea was performed. CONCLUSIONS: The following recommendations can be made for treatment of blunt testicular injury: if rupture of the testis is detected sonographically, immediate surgical exploration is indicated. Every change in testicular structure must be considered a possible sign of testicular rupture. Sonographically detected hematoceles without visible signs of rupture are not given clear recommendations in the literature. However, early surgical intervention (within 72 hours) seems to increase the likelihood of preserving the testis. In every case of unclear clinical or sonographic findings, surgical exploration of the scrotum should be done. If offers a safe and quick diaganosis with a very low complication rate. PMID- 14566691 TI - [Intratesticular cysts in infancy - case report and review of the literature]. AB - INTRODUCTION: Intratesticular cysts in neonates and infants are rare findings compared to other cystic lesions of the testes and can be diagnosed by high-power ultrasonography. In contrast to simple epithelial cysts seen in adults, which are usually small and incidental findings, intratesticular cysts in infants are often diagnosed because of an increase in scrotal size. CASE REPORT: We report the case of a 6-month-old child with painless swelling of the right scrotum and the ultrasonographic finding of an intratesticular cyst. Together with surgical enucleation of the cyst, biopsy of the macroscopically normal testicular parenchyma was performed and showed tubular atrophy in the vicinity of the cyst. CONCLUSIONS: In view of possible increase in size with resulting atrophy of testicular parenchyma, intratesticular cysts in neonates and infants should be treated surgically by simple cyst resection. PMID- 14566692 TI - [Cystinuria - Cystine Stones: Recommendations for Diagnosis, Therapy and Follow up]. AB - Cystinuria, an autosomal-recessive disorder, is the cause of 1 - 2 % of all kidney stones observed in adults and about 10 % of those observed in infants. Despite increasing understanding of underlying pathomechanisms, patients still form recurrent stones and have to undergo repeated interventions with increasing risk of renal insufficiency. Dietary and medical metaphylaxis may lower the frequency of recurrent stones but are often not practiced. Regular follow-up examinations and optimal therapy significantly enlarge stone-free intervals. This review offers an overview of the underlying pathogenetic mechanisms as well as guidance for diagnosis, monitoring, metaphylaxis and therapy of cystinuria following the recommendations of the Deutsche Gesellschaft fur Urologie (DGU) and the European Association of Urology (EAU). PMID- 14566693 TI - [The elderly patient with urge incontinence or urge-stress incontinence - efficacy and cardiac safety of propiverine]. AB - PURPOSE: The efficacy of propiverine in elderly patients suffering from urge incontinence or urge-stress incontinence was intended to be investigated. Especially in elderly patients a cardiac influence of propiverine is possible due to its dual mode of action. That is why besides the efficacy especially the cardiac safety was intended to be investigated. MATERIAL AND METHODS: Ninety eight patients (21 male, 77 female; 67.7 +/- 6.3 years of age) suffering from urgency, urge incontinence or mixed urge-stress incontinence were included in the double-blind, multicentre, placebo-controlled, randomized study. After a two-week placebo run-in period, the patients received propiverine (15 mg t. i. d.) or placebo (t. i. d.) for four weeks. Before (U 1, U 2) and during the treatment period (U 3, U 4), standard ECG's and 24 h long-term ECG's were recorded. RESULTS: Propiverine caused a significant reduction in the micturition frequency (U 2 : 8.7 +/- 4.2, U 4 : 6.5 +/- 3.2 ml; p < 0.01) reflected in a significant increase in the average micturition volume (U 2 : 163.5 +/- 65.9, U 4 : 216.3 +/- 101.5 ml; p < 0.01) and a significant reduction in the episodes of incontinence ( 54 %; p < or = 0.05). These findings were confirmed by the overall assessment after four weeks in which approximately 90 % of patients under propiverine were either free from urge incontinence and urge symptoms or improved. The efficacy parameters demonstrated a better efficacy for urge incontinence than for mixed urge-stress incontinence. Resting and ambulatory electrocardiograms evidenced no significant changes. Neither QTc interval nor other cardiac parameters were relevantly altered. The frequency of cardiac events (Lown classes IV a/b) was fortuitous, revealing no difference between placebo and propiverine. The incidence of adverse events was very low (2 % dryness of the mouth under propiverine) and confirmed by the findings from the quality of life questionnaires. CONCLUSIONS: A favourable benefit-risk ratio in the treatment of elderly patients suffering from urgency, urge incontinence or combined urge stress incontinence is therefore proven for propiverine. Cardiac arrhythmia were not induced. PMID- 14566694 TI - [Quality of life after surgical correction of penile deviation with the Schroeder Essed plication]. AB - PURPOSE: For the correction of congenital and acquired penile deviations, there are two established operative methods: the Nesbit procedure and the Schroeder Essed technique. However, which operative method is optimal with respect to the specific underlying pathology, is still controversial. The aim of the study was to evaluate patient satisfaction retrospectively after modified Essed-Schroeder plication using quality of life questionnaires. MATERIAL AND METHODS: Between June 1997 and June 2000, 59 patients underwent Essed-Schroeder at our institution for correction of a penile deviation. Mean patient age was 40 years (18 - 71) and mean follow-up 30 months (19 - 36). A standardised questionnaire was sent to all patients. Of these, the questionnaires completed by 50 patients could be assessed as valid. RESULTS: Of these 50 patients, 22 suffered from a congenital penis deviation, 28 from Peyronie's disease. The percentage of patients who were able to perform sexual intercourse pre- and postoperatively was 62 % and 90 %, respectively. The proportion of patients who felt hampered during intercourse was reduced from 68 % to 48 %. Frequency of pain occurring during intercourse was diminished by half. Sixty percent of the patients would choose to have the same operation again, while 22 % were dissatisfied with the results. None of the patients showed complete erectile dysfunction after the operation. Seventy-four percent of the patients observed a decrease in penile length. CONCLUSIONS: In patients with Peyronie's disease, the Schroeder-Essed technique with tunica plicature leds to significant improvement in relatively few cases. Alternative methods, such as the Nesbit corporoplasty, appear to be superior in these patients. In cases of congenital penile deviation, however, where the aim is cosmetic correction under maximum protection of erectile function, the Schroeder Essed tunica plicature has shown good results with regard to penile function and quality of life. PMID- 14566695 TI - [Bladder cancer and the sentinel node concept]. AB - PURPOSE: Lymph node status is one of the most important prognostic factors in muscle-invasive bladder cancer. The extent of lymphadenectomy performed in conjunction with cystectomy and the question as to whether this is a staging or therapeutic intervention are matters of discussion. The aim of this study was to evaluate the sentinel node (SN) concept and to correlate findings with tumour status in excised regional lymph nodes. MATERIAL AND METHOD: 26 patients scheduled for cystectomy were investigated with preoperative lymphoscintigraphy, peroperative dye detection (Patent Blue) and dynamic lymphoscintigraphy (Nanocoll or Albures 50 MBq/ml). The substances were injected adjacent to the tumour in the detrusor muscle. RESULTS: Sentinel nodes were detected in 21 of the 26 of the investigated patients. 7/21 SN were located outside the obturator fossa. Of the eight patients with lymph node metastasis, five displayed metastasis in lymph nodes outside the obturator fossa. There was one false negative SN in a patient with multifocal tumour, while in the other seven patients with lymph node metastasis, these were detected in the SN. CONCLUSION: Sentinel node detection is possible in most cases of bladder cancer scheduled for cystectomy. The significance of utilizing this method to detect lymph node metastasis outside the obturator fossa warrants further investigation. PMID- 14566696 TI - [Traumatic renal pelvis rupture associated with previously unknown stenosis at the renal pelvis junction]. AB - Traumatic rupture of the renal pelvis junction has been reported only rarely in the literature. Surgical exploration of the right kidney in a young man with blunt abdominal trauma showed a formerly unknown stenosis at the renal pelvis junction with hydronephrosis. Rupture of the renal pelvis by blunt abdominal trauma was related to obstruction of the ureteropelvic junction. Routine ultrasound examinations could show this pathological findings and help the patient to undergo early urological treatment. PMID- 14566697 TI - [Laparoscopic pyeloplasty]. PMID- 14566698 TI - [Penile fractures: controversy of surgical vs. conservative treatment]. AB - This paper reviews the current status of information on diagnostic and therapeutic management of penile fractures, focusing on the controversy between surgical vs. conservative treatment of this uncommon injury. Penile fracture, commonly defined as a rupture of the tunica albuginea and corpus cavernosum, is a serious urological disorder demanding surgical management. The diagnostic procedure classically consists of history taking, physical examination and urinary status. Determination of the extent of severity and location of the rupture in the tunica albuginea takes foremost priority and requires additional imaging procedures, such as ultrasound or cavernosography, which will impart enough diagnostic information for the correct choice of treatment. In some cases, the urethra is injured as well. Immediate surgical repair offers complete recovery for patients with penile fracture in most cases, even in the presence of urethral injury. Although penile fracture is easily recognized and can therefore be classified as a "first-look diagnosis", therapy remains controversial to date. The correct therapeutical approach is a vital factor in management, as the associated significant morbidity can result in complete loss of erectile function, painful erections or penis deviation. Early surgical treatment is strongly recommended because of the excellent results, shorter hospitalization, less morbidity and an early return to sexual activity. PMID- 14566699 TI - [Reconstruction of the urethral plate or ventral penile skin with foreskin or penile skin island flaps]. AB - PURPOSE: Reconstruction of urethral plate with hairless skin. Reconstruction of ventral penile skin in patients with hypospadia. MATERIAL AND METHODS: Utilization of foreskin or a circular penile island flap to form the urethral plate. Second stage urethral reconstruction with buccal mucosa. Resurfacing of the ventral surface of the penis with a foreskin island flap using the buttonhole technique. An innovation in contrast to previously described procedures is the resection of a transversal strip of dorsal penile skin to avoid bulky wedges of skin on the sides of the penis. MATERIAL AND RESULTS: The urethral plate was reconstructed in 2 hypospadia-scripples and in one patient with penoscrotal hypospadia. Urethra reconstruction with buccal mucosa was accomplished without complications in 2 patients in a second stage. Partial necrosis of the most distal port of penile skin was observed as a complication after the first operation in one patient. Functional and cosmetic results were good after treatment with split skin thickness grafts. The penis was straightened in 2 patients with hypospadis sine hypospadis and the urethra reconstructed according to Mathieu in 3 patients with distal urethral defects. The ventral skin defect was covered with an outer leaflet of foreskin island flap using the buttonhole technique. Complications were not observed postoperatively or in later follow ups. Functional and cosmetic results were good in all patients. CONCLUSION: The foreskin island flap is excellent for reconstruction of missing skin on the ventral shaft of the penis. PMID- 14566700 TI - [Bladder wall tension with physiological bladder emptying in patients with detrusor hyperactivity and infravesical obstruction]. AB - PURPOSE: The range of maximum bladder smooth muscle tension in the published literature varies between 1.3 and 59 N/cm (2) depending on the experimental setup. Based on own animal trials, we attempted to determine bladder wall tension during detrusor contraction in physiological and obstructed voiding and during detrusor instabilities in our patient collective. MATERIALS AND METHODS: In 24 patients (mean age 56 +/- 8.4 years), bladder wall tension as detrusor force per cross sectional area of bladder tissue [in N/cm (2)] was calculated following urodynamic evaluation and ultrasound determination of bladder wall thickness. The patients were divided into 3 groups with 8 patients per group as follows: group I: patients with bladder outlet obstruction in accordance with the Abrams Griffiths nomogramme; group II: patients with detrusor instabilities; group III: patients with normal bladder emptying. RESULTS: Maximum bladder wall tension in group I was 9.8 +/- 3.9 N/cm (2). During bladder instabilities (group II), maximum bladder wall tension was 11.7 +/- 2.6 N/cm (2). Maximum bladder wall tension was 2.8 +/- 0.5 N/cm (2) in group III, which was significantly lower (p < 0.05, Mann-Whitney U-Test) than that of groups I and II. CONCLUSIONS: We conclude that bladder wall tension is distinctly lower in patients with normal voiding than in those with obstructed micturition or detrusor instabilities. Furthermore, our results suggest that the detrusor is not fully activated during normal voiding and is thus capable of compensating for to a greater stress situation with more effective contractions. PMID- 14566701 TI - [Long-term follow-up of interstitial laser coagulation in the treatment of benign prostatic hyperplasia]. AB - INTRODUCTION: Besides standard open or transurethreal adenoma resection, less morbid interstitial laser coagulation (ILC) is an alternative therapeutical option that could reduce lower urinary tract symptoms, especially in high-risk patients. Although short-term results indicate effectiveness, reliable long-term statistics are still lacking. Therefore, to assess long-term satisfaction and overall success rate, we re-evaluated patients with a mean follow-up of 7 years after laser treatment. METHODS: A total of 72 patients had been included in our ILC programme between 1993 and 1995. Mean age was 74 years. About 45 % of the patients had since died of other causes. In all, 23 patients were evaluated by telephone questionnaire, International Prostate Symptom Score (IPSS), Quality of Life (QoL), second surgical interventions or medical therapy. Our patient group was treated with interstitial Nd:YAG laser coagulation (mediLas fibertom). A perineal (34 %), transurethral (23 %) or combined (43 %) approach was chosen, depending on the preoperative volume of the prostate (range 40 - 100 ml; mean 59.3 ml). RESULTS: 68.4 % of the patients were satisfied with their current urological situation. Mean IPSS was 8.8 vs. 18.8 preoperatively mean QoL 1.5 vs. 3.3. 15.8 % had undergone conventional transurethral prostatic resection in the interim; one patient uses a urine catheter. 15.8 % receive medical treatment for lower urinary tract symptoms. CONCLUSIONS: Although the results of standard TUR or open surgery imply higher success, the long-term results of ILC demonstrate effectiveness. Further follow-up studies on a larger number of patients are advisable. The low morbidity of ILC makes this procedure an interesting alternative option in the treatment of high risk patients. PMID- 14566702 TI - [Ovarian torsion--an unusual cause of childhood urinary retention]. AB - Acute urinary retention in children may be associated with a broad spectrum of different causes. Acute appendicitis or acute ovarian torsion are rare and complicate the diagnosis. The case of a 6-year-old girl with urinary retention caused by ovarian torsion is reported, in which clinical presentation showed nonspecific complaints and ultrasound demonstrated a retrovesical mass. The long standing ovarian torsion could only be diagnosed by laparotomy. This case once again demonstrates possible difficulties in obtaining a correct diagnosis, especially when imaging techniques reach their limits. Here, the anatomical structures had become so altered by adhesions of the omentum that they were no longer identifiable, rendering the advantageous, non-invasive laparoscopy ineffective. As a result, laparotomy became the most accurate and reliable method for obtaining a correct diagnosis. The occurrence of acute urinary retention with ovarian torsion can be explained by an earlier disturbance of the vegetative nervous system. PMID- 14566703 TI - [Abnormalities of the urachus]. PMID- 14566705 TI - eIF2B-related disorders: antenatal onset and involvement of multiple organs. AB - Leukoencephalopathy with vanishing white matter, also called "childhood ataxia with central nervous system hypomyelination," is the first human disease related to mutations in any of the five genes encoding subunits of eukaryotic initiation factor eIF2B or any translation factor at all. eIF2B is essential in all cells of the body for protein synthesis and the regulation of this protein synthesis under different stress conditions. It is surprising that mutations in the eIF2B genes have been reported to lead to abnormalities of the white matter of the brain only, although it has been shown recently that ovarian failure may accompany the leukoencephalopathy. Another surprising observation is that the onset of the disease varies from early childhood to adulthood, with the exception of Cree leukoencephalopathy, a disease related to a particular mutation in one of the eIF2B genes, which invariably has its onset within the first year of life. We analyzed the eIF2B genes of nine patients with an antenatal- or early-infantile onset encephalopathy and an early demise and found mutations in eight of the patients. In addition to signs of a serious encephalopathy, we found oligohydramnios, intrauterine growth retardation, cataracts, pancreatitis, hepatosplenomegaly, hypoplasia of the kidneys, and ovarian dysgenesis. Until now, no evidence had been found for a genotype-phenotype correlation, but the consistently severe phenotype in affected siblings among our patients and in Cree encephalopathy patients suggests an influence of the genotype on the phenotype. PMID- 14566707 TI - Evaluation of Web sites on management of pain in children. AB - Increasingly, consumers access the Internet for information about their health problems and treatments and to learn more about their health care. Although Web sites can be valuable resources, the information may not be accurate or current. The purpose of this project was to evaluate the quality of Web sites for parents on the management of children's pain. The Health Information Technology Institute criteria were used to evaluate 40 Web sites identified from two search engines: MSN and Google. After the evaluation process was completed, the readability of the sites was determined. Of the 40 sites, 29 (72.5%) provided useful information for parents searching to educate themselves about pain management. Other sites advertised the pain management services of their facility or were not relevant for patient education. The reading levels of the Web sites ranged from grade 7.7 to 12; the mean reading grade level of the 40 sites was 10.8, too high for many consumers. This article discusses the role of the nurse in evaluating health Web sites and teaching patients how best to use the Web for their health information. PMID- 14566704 TI - Germline inactivation of PTEN and dysregulation of the phosphoinositol-3 kinase/Akt pathway cause human Lhermitte-Duclos disease in adults. AB - Lhermitte-Duclos disease (LDD), or dysplastic gangliocytoma of the cerebellum, is an unusual hamartomatous overgrowth disorder. LDD can be familial or, more commonly, sporadic. It has been only recently recognized that LDD may be associated with Cowden syndrome (CS). Over 80% of patients with CS carry germline mutations in PTEN. It remains unclear whether all cases of LDD, even without features of CS, are caused by germline PTEN mutation and whether somatic PTEN mutation occurs in sporadic LDD. We obtained paraffin-embedded LDD lesions from 18 unselected, unrelated patients and performed mutational analysis of PTEN. Overall, 15 (83%) of 18 samples were found to carry a PTEN mutation. All individuals with mutations were adult-onset patients, but the three without mutations were diagnosed at the ages of 1, 3, and 11 years. Germline DNA was available from six adult-onset cases, and all had germline PTEN mutations. Of these six, two had CS features, one did not have CS features, and three were of unknown CS status. Immunohistochemistry revealed that 75% of the LDD samples had complete or partial loss of PTEN expression accompanied by elevated phosphorylated Akt, specifically in the dysplastic gangliocytoma cells. These data suggest that the loss of PTEN function is sufficient to cause LDD. The high frequency and spectrum of germline PTEN mutations in patients ascertaining by LDD alone confirm that LDD is an important defining feature of CS. Individuals with LDD, even without apparent CS features, should be counseled as in CS. PMID- 14566708 TI - Temperature changes, temperature extremes, and their relationship to emergency department visits and hospitalizations for sickle cell crisis. AB - Weather changes are among the proposed precursors of painful sickle cell crises. However, epidemiologic data are mixed regarding the relationship between ambient temperature and crisis frequency. To study this relationship among a local sickle cell disease population, emergency department (ED) visits and admissions were evaluated in adults with sickle cell crisis as the primary diagnosis at a major teaching hospital in a temperate climate. Official daily ambient temperatures (average for that day) were obtained from the National Climate Data Center for the days patients visited the ED or were hospitalized, and for 24 or 48 hours prior. Daily ED visit counts and admission counts were correlated with the visit/admission day's ambient temperature, with the ambient temperature 24 hours before admission, and with the magnitude of change in daily ambient temperature over the prior 24 or 48 hours. For all correlations, statistical significance was defined as a p value of <0.01 and clinical significance was defined as a moderate or greater correlation, absolute value of r >/= 0.30. ED visits or admissions correlated statistically, but not clinically, with daily temperatures. On days when temperatures were <32 degrees F or >80 degrees F, these correlations were statistically significant, but clinical significance was variable. ED visits or admissions correlated only statistically with temperatures 24 hours prior, even on days when temperatures were <32 degrees F. When temperatures were >80 degrees F, the correlations were statistically significant, but there was a reverse, clinically significant correlation between admissions and temperatures. Finally, only statistically significant correlations were found between ED visits or admissions and change in temperature over the prior 24 or 48 hours. Weak or inconsistent confirmation of a relationship was found between daily ambient temperatures and ED visits or hospital admissions for sickle cell crises. PMID- 14566710 TI - Prevalence of sleep disturbance and its relationship to pain in adults with chronic pain. AB - The study examined the prevalence and relationship between sleep disturbance and chronic pain. Research questions were: (1) What is the prevalence of sleep disturbance in adults with chronic pain, and how does this prevalence compare with healthy and insomniac adults? (2) What is the relationship between sleep disturbance and chronic pain? (3) What is the relationship of patient characteristics to sleep? This descriptive, correlational field study was done at an interdisciplinary pain clinic, sampling 99 adults, and using an 11-point pain scale and a visual analog sleep scale. For every disturbance item, more than 47% of subjects reported a score of 50 or higher, twice as high as those for healthy adults, indicating disrupted sleep. For every effectiveness item, more than 54% of subjects reported a score of 50 or less, significantly lower than for healthy adults, indicating poor sleep quality. For every supplementation item, more than 60% reported mean scores of 10 or less, indicating minimal napping, yet scores were higher than for healthy adults. For all three scales, scores were similar to the mean scores for insomniacs. Soundness of sleep showed a small but significant positive (r <.30) correlation with years of pain. Supplementation scale items were not correlated with either years of pain or pain intensity. Fragmentation was significant on the basis of gender, with men having higher scores than women. Age was a negative predictor of sleep latency. Education and age were negative predictors of the quality of sleep. PMID- 14566709 TI - A pilot study to predict success with guided imagery for cancer pain. AB - Guided imagery, as other nonpharmacologic strategies, has been demonstrated to be useful for some patients. However, no tested method exists to identify which patients are likely to benefit from this pain management strategy. This pilot study tested a model to predict success with guided imagery. Major concepts tested included imaging ability, outcome expectancy, history of imagery use, match with preferred coping style, and perceived credibility of the imagery provider. A one-group pretest-posttest design was used. A sample of 62 hospitalized cancer patients currently experiencing pain rated >/=3 on a 0 to 10 scale completed questionnaires and used an audiotaped imagery intervention. Pain outcomes examined included mean pain intensity and distress, positive and negative affect, and perceived control over pain. A path analysis was conducted using multiple regression to evaluate relationships proposed in the model. Previous history with imagery predicted outcome expectancy. Imaging ability predicted mean pain intensity, positive affect, and perceived control over pain. Outcome expectancy was not a significant predictor of any pain outcomes. Baseline status and concurrent symptoms, measured as covariates, also played a significant role in predicting outcomes. Variance explained in pain outcomes ranged from 10% to 52% (adjusted R(2) = 3% to 48%). Further exploration of model variables is warranted. Findings suggest that after considering current symptom experience, imaging ability may be a useful variable to assess in order to determine whether guided imagery is an appropriate intervention for individual patients. PMID- 14566711 TI - Intimacy and family relationships of women with chronic pain. AB - This study explored family relationships and intimacy of women experiencing chronic pain from the perspective of the women and their family members. A narrative ethnographic qualitative research design was used. Narrative and field data were analyzed using the coding procedure outlined by Strauss aided by the QSR NUD*IST 4 qualitative computer software program. Twenty-five women self identified as experiencing chronic pain, and 13 family members including 7 husbands, 1 father, 1 stepmother, and 4 children living in the home were interviewed. Three patterns of behavior emerged regarding family relationships: cycle of close involvement compensated with isolation, focus directed toward others, and loss of physical sexual intimacy. Findings of this study indicate that family members' perception is that chronic pain affects important basic relationships and both emotional and physical intimacy. A need exists to relieve some of the suffering that accompanies pain in these women and their families. Based on the study's findings, several treatment strategies are recommended. PMID- 14566715 TI - The beauty of grey. PMID- 14566717 TI - Evaluation of the treatment of idiopathic clubfoot by using the Ponseti method. AB - The initial treatment of congenital idiopathic talipes equinovarus (clubfoot) is most often nonsurgical. However, surgical treatment in the form of posteromedial release is often undertaken after failure of conservative measures. The prevalence of both immediate and long-term complications in surgically treated clubfeet has cultivated a renewed interest in nonsurgical treatment. The Ponseti method for treating clubfoot has seen a revived interest among those caring for infantile clubfeet. We report on our first 34 infants (57 clubfeet) treated by using the techniques and principles described by Ponseti. Using a standard scoring system, 54 of 57 clubfeet were successfully corrected without requiring posteromedial release. Only 2 patients (3 clubfeet) required extensive surgical correction. There were 6 relapses. In all recurrent cases, there was a lack of compliance with the straight-last shoe and foot abduction bar regimen. Based on this level of initial success, we believe that posteromedial release is no longer necessary for the majority of cases of congenital clubfeet. PMID- 14566716 TI - Peroneal tendon tears: a retrospective review. AB - Tears of the peroneal tendons are not uncommon but remain an underappreciated source of chronic lateral ankle pain. The purpose of this study was to identify the typical patient profile and nature of the injury, to analyze the course of treatment, and to determine the prevalence of complications seen with surgical repair. Forty patients with chronic pain over the peroneal tendons from the Foot and Ankle Institute at the Western Pennsylvania Hospital underwent peroneal tendon repair. During a 3-year period, a retrospective review was performed by evaluating medical records, surgical reports, and radiographs. The average patient age was 42 years (range, 13 to 64 years). The most common cause was an ankle sprain or other traumatic injury (58%). Peroneus brevis tears (35 patients; 88%), peroneus longus tears (5 patients; 13%), combined peroneus brevis and longus tears (15 patients; 37%), low-lying peroneus muscle belly (13 patients; 33%), lateral ankle ligamentous disruptions (13 patients; 33%), and peroneal subluxation (8 patients; 20%) were identified during surgery. The average follow up was 13 months (range, 9 to 40 months). Ninety-eight percent of the patients were able to return to full activities without pain at final follow-up. The minor complication rate (transient symptoms) was 20%. Clinically significant (major) complications (continued symptoms or revisionary surgery) occurred in 10% of patients. This study indicates that lateral ankle ligamentous incompetence, combined peroneal brevis and longus tears, and low-lying peroneus muscle belly commonly coexist in patients with peroneal tendon injuries. Appropriate surgical intervention of peroneal tendon tears and their coexisting pathology yields successful and predictable results with few clinically significant complications. PMID- 14566718 TI - Decompression of peripheral nerves for diabetic neuropathy in the lower extremity. AB - Consideration of the presence of a compressive neuropathy overlying an existing diabetic neuropathy in symptomatic patients and surgical decompression of these compressed nerves in selected patients is presented. The short-term results of 33 lower extremities treated with external neurolysis of the common peroneal, deep peroneal, and tarsal tunnel nerves are presented with a mean follow-up time of 3 months (range, 1 to 6 months). The surgery was performed in an attempt to relieve pain and to restore normal sensation in the foot. All procedures were performed under spinal or general anesthesia. Candidates for the procedure were type 1 or type 2 diabetics with symptomatic somatosensory neuropathy (pain, burning, tingling, and/or numbness) and preoperative computer-assisted neurosensory testing that confirmed the presence of elevated nerve threshold levels and axonal degeneration in the foot and leg. External neurolysis of the involved nerves provided good to excellent results in 90.0% of those patients with preoperative neuropathic pain, and restored sensation at good to excellent levels in 66.7% of those patients with preoperative neuropathic numbness. The mean visual analog score for pain assessment was 9.0 preoperatively and 3.2 postoperatively for those patients with pain as a symptomatic complaint (n = 30). There were 4 complications (12%) and all were early cases consisting of a noninfected wound dehiscence of the tarsal tunnel incision, which went on to heal without consequence. These initial short-term results suggest that external neurolysis of the common peroneal, deep peroneal, and tarsal tunnel nerves in selected patients with symptomatic diabetic neuropathy and an overlying compression neuropathy as determined by using computer-assisted neurosensory testing appears to be an effective treatment for providing pain relief and restoration of sensation in the foot. PMID- 14566719 TI - Radial forearm free flap for coverage of postoperative lateral heel wounds after open reduction and internal fixation of the calcaneus. AB - For intraarticular calcaneus fractures, open reduction and internal fixation has become commonplace for the reduction of morbidity of postinjury arthritis. Despite adequate surgical results, there are often associated postoperative wound complications. The purpose of this study was to describe a unique application of the radial forearm free flap for coverage of lateral postoperative heel defects seen after calcaneal fixation. Seven lateral heel wounds after open reduction of calcaneal fractures in 6 patients were covered with radial forearm free flaps. The technique used involved passage of the pedicle of the laterally placed flap anteriorly to the Achilles tendon so that it can be anastomosed to the posterior tibial artery. Flap application was 100% successful, with good functional and cosmetic results in all patients. The radial forearm free flap provides a quick, reliable, and easily harvested source of coverage for lateral heel wounds seen after open reduction and internal fixation of the calcaneus. Tunneling of the flap pedicle anterior to the Achilles tendon is simple and provides the additional advantages of access to reliable vessels, reduced scarring, and avoidance of further wound problems often seen with incisions used to access the anterior tibial artery. PMID- 14566720 TI - The oblique proximal phalangeal osteotomy in the correction of hallux valgus. AB - An oblique proximal phalangeal osteotomy is introduced as an alternative to the Akin procedure for the treatment of hallux valgus. It consists of a single oblique osteotomy directed from proximal-dorsal to plantar-distal. A retrospective review of 32 patients who underwent 36 oblique proximal osteotomies is presented. The mean follow-up was 11 months (range, 3 to 21 months). Pre- and postoperative radiographic measurements of the distal articular set angles showed an average correction angle of 12 degrees (range, 7 degrees to 22 degrees; SD = 2.99). Mean range of first metatarsophalangeal joint motion was 82% of the preoperative value when combined with distal metatarsal osteotomy, and 87% of the preoperative value without a distal metatarsal osteotomy. Time to clinical and radiographic healing averaged 5 weeks (range, 4 to 8 weeks). The average amount of phalangeal shortening was 1 mm (range, 0 to 2 mm). There were no delayed or nonunions. Subjective assessment conducted by retrospective review of 19 patients at 6 months postoperatively showed that 17 were completely satisfied (90%), 1 was satisfied (5%), and 1 fairly satisfied (5%). Nineteen would have the surgery again (1 with reservations). Eighteen were very satisfied with appearance and 1 was improved. Fourteen returned to shoe gear at weeks 6 to 8, and 5 returned at weeks 8 to 12. Eighteen were satisfied with pain relief and 1 was improved. The study indicates that the oblique proximal phalangeal osteotomy is an effective, reliable, and technically simple procedure for correction of deformities of the proximal phalanx. The advantages include minimal shortening, ease of adjustability, and a construct conducive to rigid fixation. PMID- 14566721 TI - Chondrosarcoma of the bones of the feet. AB - Twelve chondrosarcomas of the bones of the feet from 11 patients in the Scottish Bone Tumor Registry were reviewed. One patient with diaphyseal aclasis (osteochondromatosis) developed 2 chondrosarcomas. The mean age of patients was 52.3 years (range, 17 to 83 years). Men were predominantly affected. Four tumors affected the tarsal bones; the rest involved the short tubular bones. The usual clinical presentation was a painful, progressively enlarging swelling. Radiologically, most showed some bone expansion, cortical destruction with indistinct margins, and soft-tissue extension. Histologically, the majority were middle-grade tumors. Treatment included curettage or local excision for 4 tumors and amputation or ray resection for 8 tumors. Follow-up varied from 6 months to 18 years (average, 5.8 years). Local recurrence after surgery was seen in 3 patients. All 3 died because of metastases to the lungs or brain. PMID- 14566723 TI - What is the shelf life of physician-mixed antibiotic-impregnated calcium sulfate pellets? AB - This pilot study was undertaken to evaluate the short-term in vitro antimicrobial stability of both vancomycin- and tobramycin-impregnated calcium sulfate pellets mixed and stored in a clinical setting. Powdered tobramycin sulfate (500 mg) and vancomycin hydrochloride (500 mg) were blended into separate basins containing 25 g of surgical-grade calcium sulfate powder, then mixed with 8 mL of sterile saline. From this admixture, 6.0-mm pellets were produced. These were removed from the sterile container (stored at room temperature) at 1, 7, 30, 60, 90, and 120 days and tested against a variety of pathogenic bacterial isolates by using a modification of the standardized Kirby-Bauer test. Control pellets containing no antibiotic were also evaluated. There was no inhibition of bacterial growth by the non-antibiotic-impregnated (control) pellets. There was no appreciable difference in the zones of inhibition for any of the organisms with pellets stored for 1, 7, 30, 60, 90, or 120 days. Zones of inhibition for the various antibiotics to the strain of organism tested ranged from 17 mm to 30 mm, depending on the pathogenic isolate and the antibiotic evaluated. The zones of inhibition observed were similar to those designating antibiotic susceptibility in the Kirby-Bauer test. The results of this preliminary study suggest that clinician-mixed calcium sulfate pellets containing either vancomycin or tobramycin, when stored under normal room temperature and ambient humidity, appear to maintain their antimicrobial characteristics for at least 120 days. PMID- 14566722 TI - Percutaneous drilling for chronic heel pain. AB - The authors report a retrospective study involving 25 feet in 21 patients who underwent percutaneous drilling for chronic heel pain. Patients with increased activity of the heel were considered for surgical treatment if there was increased uptake on the delayed bone scans. The average follow-up was 21 months (range, 6 to 30 months). All patients were treated in day surgery with local anesthesia. Three small holes were bored in the medial cortex of the calcaneus. Clinical evaluation of the parameters of pain, walking distance, fascial tenderness, paresthesias, and ankle and subtalar joint motion were evaluated preoperatively and at final follow-up. In 7 patients, repeat bone scans were performed and 6 patients had resolution of the abnormal uptake. In 81% of feet treated, there was a favorable outcome based on a subjective scoring scale. Using a visual analog pain scale, the preoperative pain level was 8.8 (range, 4 to 10), and at latest follow-up, it was 2.4 (range, 0 to 10). These results are comparable to other available surgical methods for the treatment of recalcitrant heel pain. Less predictable results were seen in patients with rheumatic and systemic pathologies and in those diagnosed with Haglund deformity. This technique appears to be effective in the relief of intraosseous congestion and bone-marrow edema. PMID- 14566725 TI - The floating metatarsal: first metatarsophalangeal joint dislocation with associated Lisfranc dislocation. AB - The authors report a case of an irreducible dorsal dislocation of the first metatarsophalangeal joint, with concomitant Lisfranc dislocation and fractures of the second, third, and fourth metatarsals. This combination has been reported only once in the literature. This extremely rare combined injury results in a floating metatarsal. Open reduction of the metatarsophalangeal joint dislocation and fixation of Lisfranc joint and metatarsal fractures with Kirschner wires was performed. One year after surgery, the patient is active and the first metatarsophalangeal joint is asymptomatic, but there is mild pain in the Lisfranc joint. PMID- 14566724 TI - Aeromonas hydrophilia infections after penetrating foot trauma. AB - The bacterium Aeromonas hydrophila is an anaerobic gram-negative bacillus commonly found in natural bodies of water and can cause infection in patients who suffer water-associated trauma or in immunocompromised hosts. The authors present 5 cases of penetrating wound trauma that did not involve any aquatic environment and developed rapidly forming infections. All patients presented with severe pain, cellulitis, ascending lymphangitis, fever, and pain on range of motion of the joint near the traumatic site. Presentation of clinical symptoms mimicked that of a septic joint or of severe streptococcal infection. All patients required surgical incision and drainage, intravenous and oral antibiotics using levofloxacin or bactrim, and local wound care. Results from cultures taken intraoperatively showed only A hydrophilia in every case. Resolution of symptoms occurred rapidly after surgery, and clinical resolution was seen within 72 hours. Each patient healed uneventfully and returned to preinjury status. PMID- 14566726 TI - Use of the medial arch as a donor site for split-thickness skin grafts. PMID- 14566727 TI - Using a hook plate as alternate fixation for fifth metatarsal base fracture. PMID- 14566733 TI - Total joint arthroplasty in the extremely elderly: hip and knee arthroplasty after entering the 89th year of life. AB - The goal of this study was to evaluate the complications and efficacy of total joint arthroplasty in the extremely elderly and compare the survival with the normal age-matched population. One hundred one joint arthroplasties (45 total knee arthroplasties [TKAs], 56 total hip arthroplasties [THAs]) were performed in 83 patients 89 years old and older. Over an average follow-up period of 2.5 years, 26 (31%) of the patients died. Three patients (3.6%) died within the first 2 months' postoperatively. The perioperative medical complication rate (excluding deaths) was 14%. Significant improvements were noted in pain scores, Harris Hip Scores, and Knee Society Scores. The survival of patients in their nineties who undergo total joint arthroplasty is at least equal to the survival of an age matched population for 2.5 years following surgery. With careful patient selection and patient care to minimize medical complications, total joint arthroplasty can be an excellent option for patients who are age 89 and older. PMID- 14566734 TI - Surface analysis of posterior stabilized femoral components used in total knee arthroplasty. AB - The purpose of this study was to evaluate the preimplantation surface finish of the femoral component used in total knee arthroplasty. Nine femoral components from various manufacturers were obtained for analysis. All were fixed-bearing, posterior-stabilized implants intended for cemented arthroplasty. Surface characterization involved light profilometry over multiple surface points from the condylar surface, trochlear groove, and the posterior stabilizing cam. Results revealed that the condylar surface roughness was not significantly different when comparing designs with a mean roughness average ranging from 43.6 to 116 nm. The trochlear surface was generally similar to that of the condyles in most implants, although significant differences were seen in 2 implants (P <.05). Of note, the articular surface of the stabilizing cam was uniformly rougher than the condylar surface in all implants and significantly rougher (P <.05) in 6 of the 9 components. PMID- 14566735 TI - Early failure mechanisms of constrained tripolar acetabular sockets used in revision total hip arthroplasty. AB - Fifty-eight patients received an Osteonics constrained acetabular implant for recurrent instability (46), girdlestone reimplant (8), correction of leg lengthening (3), and periprosthetic fracture (1). The constrained liner was inserted into a cementless shell (49), cemented into a pre-existing cementless shell (6), cemented into a cage (2), and cemented directly into the acetabular bone (1). Eight patients (13.8%) required reoperation for failure of the constrained implant. Type I failure (bone-prosthesis interface) occurred in 3 cases. Two cementless shells became loose, and in 1 patient, the constrained liner was cemented into an acetabular cage, which then failed by pivoting laterally about the superior fixation screws. Type II failure (liner locking mechanism) occurred in 2 cases. Type III failure (femoral head locking mechanism) occurred in 3 patients. Seven of the 8 failures occurred in patients with recurrent instability. Constrained liners are an effective method for treatment during revision total hip arthroplasty but should be used in select cases only. PMID- 14566736 TI - Operative stabilization of supracondylar femur fractures above total knee arthroplasty: a comparison of four treatment methods. AB - Periprosthetic fractures of the distal femur above a total knee arthroplasty present a challenging surgical problem for orthopedic surgeons. Numerous operative and nonoperative treatment options exist including casting, Rush rods, supracondylar nails, and plate fixation. Potentially significant complications are associated with all current treatment alternatives. Plate or nail constructs frequently achieve limited distal fixation, leading to loss of fixation and varus angulation. This complication was not observed with the Less Invasive Stabilization System (LISS). In addition, our early results demonstrate the superiority of LISS treatment with low infection rates, no requirement for acute bone grafting, and secure fixation allowing for immediate postoperative mobilization. LISS fixation may offer a superior surgical treatment option for periprosthetic distal femur fractures. PMID- 14566737 TI - Brake response time after total knee arthroplasty: when is it safe for patients to drive? AB - Total knee arthroplasty (TKA) dramatically decreases pain and improves mobility and quality of life. However, little has been reported on its effect on driving ability in the early postoperative period. This prospective study was conducted to compare preoperative and postoperative brake response times (BRTs) in patients undergoing TKA for osteoarthritis. The results showed that patients returned to preoperative BRT as early as 3 weeks after surgery, and at 9 weeks after surgery, BRTs were significantly improved over baseline. Based on BRT analysis patients undergoing TKA may be allowed to return to driving 6 weeks after surgery. PMID- 14566738 TI - Hydroxyapatite-coated femoral components in total knee arthroplasty: medium term results. AB - Thirty-six consecutive total knee arthroplasties with hydroxyapatite (HA)-coated femoral components were prospectively followed up according to a standardised protocol for a mean period of 10 years (range, 7-11 years). Clinical and radiologic examination was performed at each follow-up evaluation. One patient (1 knee) was lost to follow up at 7 years, and 4 other patients (6 knees) died. Radiographs were studied for signs of loosening and presence of lucencies. Survival analysis was performed using a life table with revision as the criterion for failure. Two knees (1 patient) were revised because of aseptic loosening. The tibial component of one other knee was revised because of osteolysis in the medial tibial condyle. In the remaining knees, no radiologic evidence of loosening was seen. The survival of the femoral component at a mean follow-up time of 7 years, when 31.5 knees were at risk, was 94% with revision for femoral aseptic loosening as the end point. At 10 years, when 24 knees were at risk, the survival remained 94%. PMID- 14566739 TI - Impaction bone grafting in revision hip surgery. AB - We retrospectively reviewed the use of impaction bone grafting with cement for the fixation of femoral and acetabular components in revision hip arthroplasty. Seventy hips formed the basis of the study, with a mean follow-up time of 37 months. Sixty-eight percent of the femurs showed severe bone loss (Endoklinik grades 3 and 4). The mean Harris hip and Merle D'Aubigne Postel scores were 84 and 15.4, respectively. Massive subsidence occurred in only one femoral revision (>10 mm) and cup migration >5 mm in 6 cases. Loosening was seen in 1 revision for sepsis but none for aseptic loosening. Five complications were identified that were related to the surgical technique. We therefore support the use of this technique in revision surgery in patients with extensive bone loss. PMID- 14566740 TI - Clinical assessment of tibial polyethylene thickness: comparison of radiographic measurements with as-implanted and as-retrieved thicknesses. AB - Sixty-six posterior cruciate-retaining inserts of one fixed-bearing design were revised because of wear or osteolysis. Tibial polyethylene thickness was estimated from standard anteroposterior radiographs and compared with the original or final minimum thicknesses. The most accurate method involved measuring the shortest distance from each femoral condyle to a transverse line through the middle of the baseplate's superior surface. On radiographs acquired 6 weeks after arthroplasty, the mean error was -0.1 mm (underestimate) +/- 0.6 mm (standard deviation), with 72% of the measurements within 0.5 mm of the original minimum thickness and 87% within 1.0 mm. The method was less accurate for radiographs made before revision, for which the mean error was 0.6 mm (overestimate) +/- 1.0 mm. In this study, 41% and 70% of the prerevision measurements were accurate to within 0.5 and 1.0 mm, respectively. PMID- 14566741 TI - Knee scores change with length of follow-up after total knee arthroplasty. AB - Patients undergoing primary total knee arthroplasty (TKA) between 1989 and 1994 were evaluated preoperatively and postoperatively with the Knee Society scoring system at 3 time intervals: 1 to 2 years, 3 to 6 years, and more than 7 years. Knee prosthesis and functional scores improved statistically after TKA. At last follow-up evaluation, a statistically significant decline was found in functional knee scores that was related to an increase in numbers of patients in category C. Progression of arthritis at other sites, especially lumbar spine and hips, and cardiopulmonary problems were the most common cause of limited functional capacity. Women were found to have lower preoperative and postoperative functional scores. Patient weight, body mass index (BMI), and age at surgery showed no correlation with postoperative functional scores. The inclusion of functional categories would appear to be warranted when reporting long-term results of TKA. PMID- 14566742 TI - Patients with poor preoperative functional status have high expectations of total hip arthroplasty. AB - Recent reports indicate that patients' expectations should be assessed as part of routine practice before total hip arthroplasty (THA). A total of 1,103 THA patients preoperatively completed the Hospital for Special Surgery Total Hip Replacement Expectations Survey, an 18-item scale tested for validity and reliability. Patients also completed measures of hip-specific function (American Academy of Orthopaedic Surgeons Hip/Knee Module), and overall physical function (SF-36). Patients with worse preoperative function had higher expectations of THA and were more likely to rate their expectations as very important compared with patients with better function. In multivariate analysis, older patients, men, and those with worse functional status (both hip-specific and overall physical function) had more expectations of THA. Identifying and addressing expectations, particularly possibly unrealistic expectations, are important elements in discussions with patients before THA. PMID- 14566743 TI - Clinical outcome and incidence of pregnancy after bipolar and total hip arthroplasty in young women. AB - Female patients under 45 years of age (average, 32.5; range, 14-44) at the time of primary bipolar or total hip arthroplasty (THA) were surveyed an average 10.5 years (range, 1.7 to 20.3 years) after the index procedure. Most patients were satisfied with the pain relief and functional abilities. Of the 109 respondents (137 hips), 37 (34%) had undergone 50 additional hip procedures. Thirteen women (12%) had 19 children at an average of 3 years after hip arthroplasty. Seven women had 1 child each; 6 had 2 children; and 1 had dizygotic twins. Eleven infants were delivered vaginally and 8 by cesarean section (5 planned, 3 unplanned). Two pregnancies were considered high risk unrelated to the hip arthroplasty. Pregnancy-related complications for these patients did not differ from those in the general population. PMID- 14566744 TI - Ninety-day mortality after shoulder arthroplasty. AB - Although mortality associated with knee and hip arthroplasty has been reported, there is no information about shoulder arthroplasty. The purpose of this study was to determine the incidence and risk factors associated with perioperative mortality after shoulder arthroplasty. Between 1970 and 2000, 2,953 patients underwent shoulder arthroplasty at our institution. A retrospective review identified all patients who died within 90 days of the procedure. The 90-day mortality incidence was 0.58% (17 of 2,953). Twelve hemiarthroplasties were performed for a pathologic fracture and one for rotator cuff arthropathy. Total shoulder arthroplasties were performed for the sequelae of trauma (2), rheumatoid arthritis (1), and avascular necrosis (1). Ninety-day mortality was closely associated with the underlying diagnosis. Perioperative mortality after shoulder arthroplasty for non-neoplastic conditions is low. PMID- 14566745 TI - Early failure of a proximally cemented, distally uncemented total hip arthroplasty. AB - This investigation prospectively studied a consecutive series of 22 proximally cemented, distal cementless, femoral components with a minimum follow-up time of 24 months. The prosthesis was a titanium, proximally cemented, femoral component with a distal press-fit fluted design. Five hips (23%) developed aseptic mechanical loosening at an average of 46.2 months and required revision. The technique required excessive removal of cancellous bone leading to a suboptimal bone-cement interface. The early failure rates of this prosthesis are unacceptably high for a contemporary hip design. To the best of our knowledge, no clinical studies have reported on the stability and longevity of this new proximal cementing technique incorporated with a distal press-fit femoral design. PMID- 14566746 TI - Influence of the infrapatellar fat pad resection in a synovectomy during total knee arthroplasty in patients with rheumatoid arthritis. AB - Influence of the infrapatellar fat pad resection in a synovectomy during total knee arthroplasty (TKA) was evaluated in patients with rheumatoid arthritis (RA). Our findings for 120 RA patients at 28 to 38 months after surgery showed that (i) a significant decrease in the number of patients with anterior aching discomfort despite a lower-lying patella was seen in patients with infrapatellar synovectomy compared with patients without infrapatellar synovectomy, and (ii) an increase in the number of patients with anterior aching discomfort, significant limited motion, slight quadriceps weakness, and significant shortening of patellar tendon length and patella height were noted among patients with infrapatellar synovectomy, including fat pad resection, than in patients without infrapatellar synovectomy. PMID- 14566747 TI - Management of periprosthetic femoral fractures. AB - We followed up 37 periprosthetic fractures to assess the factors related to the outcomes and the loosening of prostheses. The average union time is 3.1 months (range, 2-6.2 months). The most common types were fractures around the stem tip and displaced (89.1%). In our study, the loosening rate (32.4%) was low compared with other reports. These might be due to the cementless stem, but the difference was not significant (P>.05). After management of the fracture with a plate, we noted 5 complications: 2 nonunion and 3 refractures. The causes of these complications were related to screw in the level of the stem tip. Therefore, in the management of fractures with plates, we do not recommend using a screw in the area of the stem tip but rather using a cerclage proximally and bone graft. If stem was unstable or stability was uncertain, and the arthroplasty did not function well previously, revision with a distal fluted stem showed an early bony union. PMID- 14566748 TI - Cortical and cancellous morselized allograft in acetabular revision total hip replacement: minimum 5-year follow-up. AB - Sixty patients who had had revision acetabular components with impaction morselized bone graft were studied in a retrospective, matched-pair analysis. In 30 patients, the morselized graft was derived from cortical bone (group A), and in 30 patients, the graft was derived from cancellous bone (group B). The patients were matched for age, gender, weight, comorbid conditions, Charnley category, severity of bone loss, and allograft preparation. At the time of follow up, a mean of 7.4 years (range, 5-9 years) after surgery, the mean Harris Hip Scores in groups A and B were 84 and 77, respectively. Cup migration of 5 to 10 mm were noted in two patients in each group. Cup migration >10 mm was noted in 2 patients from group B and none in group A. Two patients in group A and 5 patients in group B underwent cup rerevision as a result of aseptic loosening. There was a correlation between progressive radiolucent lines extending 2 mm, cup migration, and clinical outcome. The current study showed that early and midterm clinical and radiologic outcomes such as cup migration >5 mm and radiolucent lines were significantly better using cortical morselized allograft. However, further mechanical and histologic evaluations should be performed that demonstrate the difference between these 2 types of morselized allograft. PMID- 14566749 TI - Four-angle radiographic assessment of cement mantle thickness in cemented total hip arthroplasty. AB - In this study, the cement mantle thickness of 57 hips undergoing primary cemented total hip arthroplasty (THA) were investigated with 4 angle radiographs. All surgeries were performed with a standardized posterolateral surgical approach. In addition to conventional anteroposterior and lateral radiographs, obturator oblique and iliac oblique radiographs were taken. In the evaluation of the cementing grade on the anteroposterior radiograph, the incidence of hips with thin cement mantles was 25%. In evaluating the cementing grade on the anteroposterior and lateral radiographs, we found an incidence of hips with thin cement mantles of 37%. From the evaluations with 4 angle radiographs, 10 hips were additionally assigned as being of a poor cementing grade (Mulroy's grade C 2). Thin cement mantles were mainly seen on the iliac oblique radiographs. PMID- 14566750 TI - Early head penetration into cemented Hylamer Ogee socket. AB - The authors reviewed 18 consecutive cemented total hip arthroplasties with the Hylamer Ogee socket. The average follow-up period was 47 months. All Hylamer Ogee sockets were sterilized by gamma irradiation in air, and the average period from production to surgery was 12 months. Two-dimensional penetration of the femoral head into the Hylamer Ogee socket was determined from anteroposterior radiographs of the pelvis using a digitizer. The average head penetration rate of the present series was 0.36 mm/y. Osteolysis around both components was found in 6 cases (30%). Of 4 cases defined as aseptic loosening, 2 were revised during the follow up period. To detect component loosening because of catastrophic high wear of the cemented Hylamer Ogee socket as early as possible, precise radiographic follow-up is essential. PMID- 14566751 TI - Sixteen-year follow-up of the cemented spectron femoral stem for hip arthroplasty. AB - Clinical and radiographic follow-up was performed on a consecutive series of 105 patients who underwent 120 total hip arthroplasties at the authors' institution from 1983 to 1988 with a straight, cobalt-chrome femoral stem implanted using a second-generation cementing technique. The mean age at the time of surgery was 68.5 years, and the mean follow-up was 16 years. At 16 years' follow-up, the prevalence of revision for aseptic loosening of the Spectron femoral component was only 4.2%; 5 stems were revised for aseptic loosening at a mean of 10.2 years after implantation. Sixteen-year survivorship of the component was 93.9% +/- 2.7% when revision for aseptic loosening was taken as the endpoint or 90.3% +/- 4.4% when either revision for aseptic loosening or radiographic evidence of loosening was taken as the endpoint. PMID- 14566752 TI - Dissemination of metals from a failed patellar component made of titanium-base alloy. AB - Complications related to the patella are the most common problems in total knee arthroplasty and major reasons for revision surgery. Among patellofemoral problems, metal-to-metal contact between patellar and femoral components is the worst. We measured levels of titanium, aluminum, and vanadium in serum and urine samples and compared these measures from cases in which metal-to-metal contact of the patellar component occurred with those in cases in which only polyethylene wear occurred. Serum levels of the metals in cases with metal-to-metal contact were over 10 times higher than those in cases without metal-to-metal contact. This suggests that different ranges of serum metal ion levels could eventually be correlated with a variety of mechanisms of patellar failure. Therefore, cases with metal-to-metal contact should be distinguished from cases without metal-to metal contact. These should not be considered as a single group in patellar component failure. PMID- 14566753 TI - Mechanical performance of ceramic acetabular liners under impact conditions. AB - Although new generation alumina ceramics have exhibited a reduced incidence of fracture, concern still persists about the behavior of ceramic acetabular liners under impact conditions. The objective of this study was to explore whether fracture of a new generation alumina ceramic liner was likely to occur in vivo. Ceramic liners were impacted with forces of 23, 21, 15, and 12 kN (n = 3 at each force). At 23 kN, all 3 ceramic liners fractured on the first impact; at 12 kN none of the ceramic liners fractured after 20 impacts. The threshold force of 12 kN is large in comparison with estimated physiologic forces on the hip during falls or stumbling, suggesting that ceramic liner fracture is not a definite consequence of liner impact. PMID- 14566754 TI - Fracture of the polyethylene tibial post in a posterior cruciate-substituting total knee arthroplasty mimicking patellar clunk syndrome: a report of 5 cases. AB - Fracture of the tibial post in posterior cruciate-substituting total knee arthroplasty has been described in several reports. Additionally, wear of the cam and post mechanism, including polyethylene debris generation and osteolysis of the peri-implant bone stock as been described. Users of a cam and post type posterior stabilized total knee arthroplasty are also familiar with the rare occurrence of the "patellar clunk syndrome," in which suprapatellar fibrous nodular scar tissue becomes entrapped in the intercondylar box of the femoral component as the knee extends from the flexed position and produces a palpable and sometimes audible "clunk." This condition is easily managed with arthroscopic excision of the scar tissue. A small series of 5 patients who presented with symptoms of a patellar clunk syndrome but who in fact had a fracture of the tibial post causing subluxation of the femur on the tibia is presented. The diagnostic characteristics that will help differentiate between the 2 problems is highlighted. PMID- 14566755 TI - Ender nailing for supracondylar fracture of the femur after total knee arthroplasty: five case reports. AB - We studied 5 women with supracondylar fractures after total knee arthroplasty (TKA). The mean age at fracture was 67.4 years. Four patients had chronic rheumatoid arthritis, and 1 had osteoarthritis. The range of motion, knee score, femorotibial angle, and component alignment were investigated. Flexion was slightly decreased after Ender nailing, but extension only changed in 1 patient. The knee score decreased, but all patients were ambulatory. The femorotibial angle changed in all patients. Measurement of femoral component alignment showed a postoperative change of angle alpha (1 degrees to 11 degrees ) and angle gamma (0 degrees to 9 degrees ). Union was achieved in all patients. Ender nailing is an acceptable method of treatment for supracondylar fractures of the femur after TKA if the nails can be inserted deeply into the femoral condyles. PMID- 14566761 TI - Uremic malnutrition-inflammation syndrome in chronic renal disease: a pathobiologic entity. PMID- 14566762 TI - The extent of uremic malnutrition at the time of initiation of maintenance hemodialysis is associated with subsequent hospitalization. AB - OBJECTIVE: End-stage renal disease (ESRD) patients with signs of uremic malnutrition at the time of initiation of maintenance hemodialysis (MHD) are likely to remain malnourished over the subsequent year. Because poor nutritional status is associated with worse clinical outcomes in MHD patients, we hypothesized that ESRD patients with evidence of uremic malnutrition at the time of initiation of MHD would have more hospitalization events compared with patients initiating MHD without signs of malnutrition during the first year of therapy. DESIGN/INTERVENTION: This was an observational cohort of incident MHD patients, with no specific nutritional intervention. SETTING: Vanderbilt University Outpatient Dialysis Unit. PATIENTS: All newly initiated MHD patients at Vanderbilt University Outpatient Dialysis Unit were recruited for study purposes. A total of 149 patients were included in the study. MAIN OUTCOME MEASURE: The following parameters were recorded at the time of initiation of MHD: age; race; gender; serum concentrations of albumin, creatinine, cholesterol, and transferrin; and whether the patient had insulin-dependent diabetes mellitus. The number of hospital admissions and length of stay in the hospital were recorded for all study patients during the first year of MHD. Associated hospital charges were obtained for a subgroup of 52 patients. RESULTS: Study variables were associated with hospitalization in the subsequent year, the number of hospital admissions, and the length of stay in the hospital. Patients who initiated MHD in the lowest quartile of serum albumin had a significantly greater average of admissions compared with patients who initiated in the highest quartile (1.77 +/- 1.82 versus 0.72 +/- 0.96 admissions, P =.002). The length of stay in the hospital was also higher in the lowest quartile of serum albumin (8.96 +/- 9.96 versus 3.83 +/- 5.68 days, P =.006). Serum creatinine was also inversely associated with greater average number of admissions (2.27 +/- 2.41 versus 0.83 +/- 1.68 admissions, P =.004) and longer length of stay (12.43 +/- 15.15 versus 4.72 +/- 11.57 days, P =.017) in lowest compared with the highest quartile. In addition, the costs associated with hospitalizations were higher in the group of patients initiating MHD with lower concentrations of serum albumin and serum creatinine. CONCLUSIONS: In this study of incident MHD patients, the concentrations of 2 nutritional parameters, serum albumin and serum creatinine at the time of initiation of MHD, were significantly and negatively associated with hospitalization events. There was also a trend for greater hospital charges in patients with lower concentrations of serum albumin and creatinine. PMID- 14566763 TI - A randomized controlled trial of an educational intervention to improve phosphate levels in hemodialysis patients. AB - OBJECTIVE: To determine the effect of a dietetic educational intervention on phosphate and calcium levels of hemodialysis patients. DESIGN: Parallel-group randomized controlled trial. SETTING: Teaching hospital hemodialysis unit in London, England. PATIENTS: Fifty-six stable adult hemodialysis patients with hyperphosphatemia. INTERVENTION: An educational intervention and one-to-one teaching session given by a renal dietitian, attempting to improve patients' knowledge of phosphate management and their compliance with diet and medication. OUTCOME MEASUREMENT: Patients' serum phosphate, calcium, and calcium x phosphate products in the 3 months after the intervention, compared with those before the intervention. Results were also compared with a control group that had not undergone the intervention. RESULTS: In the intervention group, serum phosphate was significantly reduced after the education session, as compared with the results previously. In the control group, there was no significant change in serum phosphate level. The improved results were sustained over a period of 3 months. Serum calcium increased in the intervention group, but this result was not significant. There was an improvement in the calcium-phosphate product in the intervention group, but again this was not significant. CONCLUSION: Dietetic educational intervention can favorably alter patients' serum phosphate levels, with potential impact on morbidity and mortality. PMID- 14566764 TI - Differences in perceptions by stage of fluid adherence. AB - OBJECTIVE: The purpose of this study was to determine in a group of hemodialysis patients whether perceptions of barriers to and benefits of adherence to fluid limitations, perceived seriousness of and susceptibility to the consequences of nonadherence, self-efficacy, and thirst differed by stage of fluid adherence. DESIGN: Cross-sectional descriptive design. SETTING: There were 147 participants from rural, suburban, and urban outpatient hemodialysis units. RESULTS: Significant differences were found in perceived benefits, barriers, seriousness, susceptibility, and thirst intensity by stage of fluid adherence. There were no significant differences in self-efficacy by stage of fluid adherence. CONCLUSION: Stage of fluid adherence may be important to consider when designing interventions to reduce fluid intake. PMID- 14566765 TI - Body composition of patients on a very low-protein diet: a two-year survey with DEXA. AB - BACKGROUND: It has been reported that patients on a very-low-protein diet (VLPD) maintain a satisfactory nutritional status because of a conserved adaptive metabolic response. However, only few studies have examined the course of nutritional status and body composition in the long term (2 years). METHODS: Thirteen stable patients (8 men; age, 55 +/- 12 years; glomerular filtration rate (GFR), 15 +/- 5 mL/min) receiving a VLPD (0.3 g/kg/day protein) supplemented with amino acids and ketoanalogues (SVLPD) were studied for 2 years. A joint visit with a physician and a dietitian and routine blood and urine analyses were performed every month. Dual-energy x-ray absorptiometry (DEXA), which was used to assess modification of body composition, and GFR (urinary 51Cr-EDTA) and urinary urea and creatinine excretion, which were used to assess nutritional status and compliance to the diet, were assessed every 3 months. RESULTS: GFR, albumin, and prealbumin levels remained stable. Urea urinary excretion decreased at 3 months and then slightly increased at 2 years, but the calculated protein intake remained low at 0.38 +/- 0.1 g/kg/day. Energy intake remained close to 30 kcal/kg/day. No significant change was observed for total fat mass or percent fat mass. After an initial decrease, lean body mass stabilized at 6 months and then increased significantly from 6 to 24 months (P =.02, paired t-test); the mean increase during this period was of 2 kg, that is, 4.6%. Urinary creatinine excretion showed the same profile. Total bone mass, lumbar or hip site bone mass, and Z-score significantly decreased from T0 to 1 and 2 years (P <.05). CONCLUSION: This study confirms that a supplemented VLPD is nutritionally safe for a long period, but attention must be paid to bone mass. PMID- 14566766 TI - Calcium-containing phosphate binder use associated with accelerated atherosclerotic coronary calcification. AB - OBJECTIVE: To report parameters and outcomes of phosphorus management in a maintenance hemodialysis patient who experienced atherosclerotic coronary calcification leading to myocardial infarction; to evaluate the role that high calcium intake over 6 years may have played in his coronary artery disease before the dangers of excess calcium intake in phosphorus management were recognized; and to describe an optimized therapeutic approach that provided improved mineral control. DESIGN: Case study. SETTING: A large outpatient in-center hemodialysis treatment unit. MAIN OUTCOME MEASURES: Serum calcium, serum phosphorus, Ca x P product. RESULTS: The year before his cardiac event, the patient's mean serum calcium was 9.6 mg/dL, mean serum phosphorus was 5.9 mg/dL, and mean Ca x P product was 57 mg2/dL2. Serum calcium peaked at 10.8 mg/dL shortly before his cardiac event. Treatment phases included high-dose calcium acetate (10 g Ca/day), low-dose calcium acetate (4 g Ca/day), low-dose calcium acetate plus low-dose sevelamer hydrochloride (4 g Ca/day plus 2.4 to 4.8 g sevelamer/day), and low dose calcium acetate plus higher-dose sevelamer hydrochloride (4 g Ca/day plus 4.0 to 12.0 g sevelamer/day). With calcium acetate and sevelamer doses optimized, serum phosphorus levels and Ca x P products continued decreasing, with mean values of 5.99 mg/dL and 50.7 mg2/dL2, respectively, and serum calcium remained stable at a mean of 8.5 mg/dL. The patient has had no further myocardial infarctions. CONCLUSIONS: This case illustrates how our growing understanding of phosphorus management and the addition of a calcium-free binder to our therapeutic armamentarium have improved phosphorus and calcium balance, reducing the risk of cardiovascular calcification. PMID- 14566767 TI - Do we need a nutrition-specific quality of life questionnaire for dialysis patients? AB - Food, eating, and quality of life are intimately related. Available generic and disease-specific health-related quality of life (HRQOL) instruments address nutrition only with respect to the ability to eat and omit more dynamic components of nutrition such as sensory enjoyment of food and food intake. For improved assessment and monitoring of nutrition-related quality of life in hemodialysis patients, a nutrition-specific questionnaire is recommended, consisting of two tools: the Appetite and Dietary Assessment Tool (ADAT), already used in many dialysis clinics, and the nutrition-specific Food Enjoyment in Dialysis (FED) questionnaire which asks 10 additional questions on sensory changes, thirst, gastrointestinal symptoms, side effects from medications and/or dialysis, and feelings of satisfaction and control. This nutrition-specific information should permit comprehensive evaluation and monitoring of changes in nutrition related quality of life. When coupled with generic and disease specific tools, it will provide a complete picture of quality of life in hemodialysis patients. PMID- 14566768 TI - Patient education. Tips for cholesterol control in patients on hemodialysis. PMID- 14566778 TI - How to build an adapted and bioactive cell microenvironment? A chemical interaction study of the structure of Ca-alginate matrices and their repercussion on confined cells. AB - Alginates are increasingly being used as medical materials (matrices for tissue regeneration, surgical sponges, hemostatic bandages, microbial and cell encapsulation, artificial bacterial biofilms, etc.). The constitution of alginate gel networks is a complex phenomenon. A great number of different kinds of polysaccharidic frameworks can come to existence depending on the conditions used for their attainment. For instance, the degree of heterogeneity and porosity of Ca-alginate beads rely on this molecular organization. The formation of structural irregularities (superficial crust, cavities, shafts, dense or light gel frameworks, ordered or chaotic domains, etc.) within the alginate gel beads are inherent to this skeletal design. Several specific staining molecules (e.g. calcon carboxylic acid, murexide, methylene blue) that are negatively or positively charged interact with the gel network. These molecules allowed us to reveal a great variety of chemical interactions shown by the pattern coloration of the internal structure of the gel. The results observed are very different for the several matrices analyzed, which could explain to a great extent the singular behavior that cells confined in these kind of matrices exhibit. PMID- 14566779 TI - A biodegradable polyurethane-ascorbic acid scaffold for bone tissue engineering. AB - A novel, nontoxic, biodegradable, sponge-like polyurethane scaffold was synthesized from lysine-di-isocyanate (LDI) and glycerol. Ascorbic acid (AA) was copolymerized with LDI-glycerol. Our hypothesis was that the AA-containing polymer foam would enhance the biological activity of the osteoblastic precursor cell (OPCs). The LDI-glycerol-AA matrix degraded in aqueous solution to the nontoxic products of lysine, glycerol, and AA. The degradation products did not significantly affect the solution pH. The physical properties of the polymer network supported the cell growth in vitro. Mouse OPCs attached to the polymer matrix and remained viable. OPCs produced multilayered confluent cultures, a characteristic typical of bone cells. Furthermore, AA release stimulated cell proliferation, type I collagen, and alkaline phosphatase synthesis. Cells grown on the LDI-glycerol-AA matrix also showed an enhancement of mRNA expression for pro-alpha1 (I) collagen and transforming growth factor-alpha1 after 1 week. Data were tested for significance with an analysis of variance model and multiple comparison test (Fisher's Protected Least Significant Difference) at p < or = 0.05. The observations suggest that AA-containing polyurethane may be useful in bone tissue engineering applications. PMID- 14566781 TI - Electrical polarization of bioactive glass and assessment of their in vitro apatite deposition. AB - We evaluated the electrical polarizability of a bioactive glass of the 45S5 type (BG) by thermally stimulated depolarization current (TSDC) measurement based on the effects of the surface charges generated by an electrical polarization treatment on the growth of calcium phosphate crystals in simulated body fluid (SBF). From the results of the TSDC measurements, the polarized BG showed broad peaks in the depolarization current density spectra. The stored electrical charges calculated from the TSDC measurements of the BG polarized with an electrical dc field of 10 V x cm(-1) at 500 degrees C for 1 h was 8.5 mC x cm(-2) and depended on the electrical dc field intensity. The depolarization of the BG was assumed to be due to a migration of sodium ions and influenced by transitions of the BG structure. The immersion tests with the SBF showed that the effects of the electrical polarization produced morphologic changes in the calcium phosphate deposit grown on the BG after the short period of 2 h. We expected the effects to be caused by a change in the ion concentrations in the surrounding SBF due to the surface charges of the polarized BG. The change in the ion concentrations induced the difference in the calcium phosphate deposition. Consequently, we confirmed that BG had an electrical polarizability, a possibility of storing large charges, and differences in the growth of the calcium phosphate deposit due to the effects of the surface charges. PMID- 14566780 TI - Effect of increasing silver content in phosphate-based glasses on biofilms of Streptococcus sanguis. AB - Silver is a powerful antibacterial ion that may be useful for dealing with localized infections, such as periodontitis. However, the use of silver in this role could be significantly improved by the development of an effective means of delivery. Phosphate-based glasses may provide a means of delivering the ions in a controlled manner. In our study, we have examined the effect of increasing silver content in phosphate-based glasses on biofilms of Streptococcus sanguis. Glasses of nominally the same dissolution rate were doped with silver at 1, 5,10, and 15 mol % and the number of colony-forming units (CFUs) determined after 6 and 24 h in a constant depth film fermenter (CDFF). The 1 mol % silver showed little change between 6 and 24 h. However, only 0.5 log CFUs were present on the glass containing 5 mol % at 6 h, and this reduced to virtually zero at 24 h. Few viable bacteria were found on the 10 mol % glass at both 6 and 24 h. The 15 mol % glass was investigated in this experiment, but no viable counts were detected. In a second set of experiments, glasses with 10 and 15 mol % silver were tested in the CDFF for up to 192 h. For the 10 mol % silver glass, there was approximately 0.8 log CFUs on this glass, which dropped to almost zero at 50 h. This was approximately 1.5-2.0 log reduction in CFUs compared to controls, and this difference was maintained for the first 50 h. After 50 h, there was a slow increase in the CFUs on all samples. However, CFUs on the 10 mol % silver glass were still suppressed up to 192 h compared to the controls. However, both controls also exhibited a decrease in viable counts at 50 h; this may have been due to carryover of silver into the control sample holders. However, this was minimized by the specimen layout in the CDFF and by having gaps between specimen sets. For the 15 mol % silver glass, counts for both this glass and the controls decreased to virtually zero between 24 and 48 h, but the numbers slowly increased up to 170 h, but the number of CFUs was suppressed compared to the 10 mol % glass at the same time point. The decrease seen is clearly the effect of the silver ions; however, the slow increase in CFUs may be accounted for by the biofilms forming thick layers on top of the glass discs inhibiting the release of ions from the glass by forming a "sacrificial layer" through which further ions have to diffuse. PMID- 14566782 TI - Modification of Ti6Al4V surfaces using collagen I, III, and fibronectin. I. Biochemical and morphological characteristics of the adsorbed matrix. AB - Studies in developmental and cell biology have established the fact that responses of cells are influenced to a large degree by morphology and composition of the extracellular matrix. Goal of this work is to use this basic principle to improve the biological acceptance of implants by modifying the surfaces with components of the extracellular matrix (ECM). Aiming at load-bearing applications in bone contact, in this study the modification of titanium surfaces with the collagen types I and III in combination with fibronectin was undertaken; fibrillogenesis, fibril morphology and adsorption of type I, III and I/III cofibrils onto titanium were assessed. Increasing the collagen type III amount resulted in a decrease of fibril diameter, while no significant changes in adsorption could be detected. The amount of fibronectin bound to the heterotypic fibrils depended on fibrillogenesis parameters such as ionic strength or concentration of phosphate, and varied with the percentage of integrated type III collagen. PMID- 14566783 TI - Modification of Ti6AL4V surfaces using collagen I, III, and fibronectin. II. Influence on osteoblast responses. AB - Responses of osteoblastic cells are influenced by morphology and composition of the extracellular matrix, and this fact has been used to improve the biological acceptance of implants by modifying the surfaces with components of the extracellular matrix (ECM). In this study, the effect of the collagen types I and III on adhesion, proliferation, and differentiation was studied, using primary osteoblastic cells from rat calvariae. Differences in alkaline phosphatase activity (ALP) and collagen synthesis were observed between differently composed collagen coatings. An increase in collagen type III resulted in an increase in collagen synthesis and a concomitant decrease in ALP activity and Ca deposition. Initial adhesion mechanism of the cells depended on the substrate (titanium, collagen, fibronectin). PMID- 14566784 TI - Oxygen radical production in neutrophils interacting with platelets and surface immobilized plasma proteins: role of tyrosine phosphorylation. AB - The interaction between neutrophil granulocytes and platelets is considered to play an important role in the inflammatory process induced by an implanted foreign material. However, the cellular mechanisms involved remain incompletely understood. We used a luminol-dependent chemiluminescence (CL) technique to analyze the generation of reactive oxygen species (ROS) in human neutrophils interacting with different plasma protein-coated surfaces in the presence or absence of unstimulated or stimulated platelets. The role of tyrosine phosphorylation in the regulation of NADPH oxidase activity was evaluated with quantitative fluorescence microscopy and the specific tyrosine kinase inhibitor genistein. We found that the ROS-production is 2 to 3 times higher in neutrophils on immunoglobulin G (IgG)-coated surfaces than in cells interacting with albumin- or fibrinogen-coated surfaces. Incubation with superoxide dismutase and catalase revealed that about 45% of the ROS was released extracellularly on IgG surfaces whereas corresponding values were 90% and 85% in neutrophils interacting with albumin and fibrinogen, respectively. The presence of platelets markedly increased the extracellular generation of ROS, mainly in neutrophils interacting with IgG- or fibrinogen-coated surfaces whereas the intracellular production was only modestly affected. Quantitative fluorescence microscopy of neutrophils stained with FITC-conjugated anti-phosphotyrosine antibodies showed a correlation between tyrosine phosphorylation, cell spreading, and ROS production. Platelets markedly amplified the anti-phosphotyrosine staining on both fibrinogen- and IgG coated surfaces whereas the low level of tyrosine phosphorylation in neutrophils on albumin-coated surfaces was not further elevated by platelets. Furthermore, the tyrosine kinase inhibitor genistein inhibited both extra- and intracellular ROS production in neutrophils regardless of the presence of platelets. We demonstrate that plasma protein coating and the presence of platelets are crucial for the inflammatory response of adhering neutrophils and that the oxidative response correlates with the extent of tyrosine phosphorylation of proteins in focal contacts. PMID- 14566785 TI - Protein adsorption and smooth muscle cell adhesion on biodegradable agmatine modified poly(propylene fumarate-co-ethylene glycol) hydrogels. AB - We synthesized positively charged biodegradable hydrogels from poly(propylene fumarate-co-ethylene glycol) block copolymer and agmatine-modified poly(ethylene glycol)-tethered fumarate by radical crosslinking, and investigated the effect of the guanidino group of agmatine on vascular smooth muscle cell adhesion and protein adsorption to the hydrogels. In the presence of serum, the number of adherent smooth muscle cells per unit surface area increased dose-dependently from 15 to 75% of the initial seeding density at 20 h as the initial agmatine modified monomer content increased from 0 to 200 mg/g. Cell spreading also depended on the initial monomer content. In the absence of serum, the number of adherent cells per unit surface area increased slightly from 10 to 17% of the initial seeding density as the initial monomer content increased from 0 to 200 mg/g. Cell adhesion increased significantly by adding exogenous vitronectin to serum-free medium, whereas exogenous fibronectin addition did not enhance cell adhesion. The enzyme-linked immunosorbent assay of fibronectin and vitronectin adsorbed onto the hydrogels revealed that the incorporation of positive charges into the hydrogels enhanced vitronectin, but not fibronectin, adsorption significantly. These results suggest that the guanidino group of agmatine enhanced cell adhesion by promoting the adsorption of serum components, and vitronectin may be one of the components. PMID- 14566786 TI - Optimal heparin surface concentration and antithrombin binding capacity as evaluated with human non-anticoagulated blood in vitro. AB - Contact between blood and a biomaterial surface takes place in many applications and is known to activate the coagulation and complement systems. Heparin surface coatings have been shown to reduce blood activation upon contact with artificial surfaces. To establish the optimal heparin surface concentration, blood was incubated in a tubing loop model at 37 degrees C. The tubing was coated with different surface concentrations of heparin and rotated at three different velocities. We demonstrate that the blood compatibility of a surface with regard to coagulation, complement, and platelet activation can be improved by increasing the heparin surface concentration in the 6-12 pmol antithrombin/cm2 concentration interval. The binding of factor H is not influenced by the increased heparin surface concentration, suggesting that this factor is not the primary regulator of complement on heparin surfaces. In addition, the heparin coating has no effect on the complement activation that occurs on gas surfaces in extracorporeal circuits. PMID- 14566787 TI - Bridging a 30-mm nerve defect using collagen filaments. AB - This article describes a 30-mm regeneration of severed peripheral nerve axons along collagen filaments. Two thousand or 4000 31-mm-long collagen filaments were grafted to bridge a 30-mm defect of the rat sciatic nerve. A collagen tube was grafted as a control. The mean number and mean fiber diameter of regenerated myelinated axons were 330 +/- 227 and 2.7 +/- 0.9 microm in the distal end of the 2000 collagen-filaments nerve guide, and 564 +/- 275 and 2.5 +/- 1.1 microm in the distal end of the 4000 collagen-filaments nerve guide at 12 weeks postoperatively, whereas in the distal end of the collagen tube, no regenerated axon was found. These results suggest that the collagen filaments guide axons of the rat's sciatic nerve to regenerate for 30 mm and act as a scaffold for axonal regeneration. Thirty-millimeter nerve regeneration of a 1-mm-diameter rat sciatic nerve by an artificial nerve guarantees a clinical application of the implant which should be very important for patients and surgeons. PMID- 14566788 TI - Interactions of antithrombin and proteins in the plasma contact activation system with immobilized functional heparin. AB - The interactions of antithrombin (AT) and the contact phase clotting factors with two commercially available heparinized surfaces are reported. The Carmeda (CBAS) and Corline surfaces along with controls (a sulfonated polyethylene surface and a CBAS analog in which the heparin used was devoid of specific AT-binding sequences) were exposed to human plasma. Adsorbed proteins were eluted and examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. The CBAS and Corline surfaces adsorbed large amounts of AT, whereas adsorption on the controls was negligible. Immunoblots for the four contact phase clotting factors indicated less contact activation on the CBAS and Corline surfaces than on the controls. Determination of adsorbed functional AT using a FXa inhibition assay showed that the CBAS surface adsorbed about 4 times as much AT as the Corline surface. Adsorption of AT to the control surfaces was minimal. Assays for adsorbed FXII and FXIIa based on kallikrein generation showed that all four surfaces adsorbed similar amounts of FXII. However, on the controls, most of the FXII was in activated form, whereas on the CBAS and Corline surfaces very little activation occurred. PMID- 14566789 TI - On the fracture of human dentin: is it stress- or strain-controlled? AB - Despite substantial clinical interest in the fracture resistance of human dentin, there is little mechanistic information in archival literature that can be usefully used to model such fracture. In fact, although the fracture event in dentin, akin to other mineralized tissues like bone, is widely believed to be locally strain-controlled, there has never been any scientific proof to support this belief. The present study seeks to address this issue through the use of a novel set of in vitro experiments in Hanks' balanced salt solution involving a double-notched bend test geometry, which is designed to discern whether the critical failure events involved in the onset of fracture are locally stress- or strain-controlled. Such experiments are further used to characterize the notion of "plasticity" in dentin and the interaction of cracks with the salient microstructural features. It is observed that fracture in dentin is indeed locally strain-controlled and that the presence of dentinal tubules does not substantially affect this process of crack initiation and growth. The results presented are believed to be critical steps in the development of a micromechanical model for the fracture of human dentin that takes into consideration the influence of both the microstructure and the local failure mode. PMID- 14566790 TI - Biologic significance of surface microroughing in bone incorporation of porous bioactive glass implants. AB - A novel chemical etching method was recently developed to create a controlled microrough surface on porous bioactive glass implants. Our earlier in vitro studies showed enhanced attachment of osteoblast-like MG63 cells on a microrough bioactive glass surface. The purpose of our current study was to confirm the in vivo significance of surface microroughening for bone bonding of bioactive glass. Porous bioactive glass cones made of sintered microspheres were surgically implanted in the anterior cortex of rabbit femurs. Peripheral quantitative computed tomography (pQCT), biomechanical push-out testing, histomorphometry, and electron microscopy (BEI-SEM) were used to analyze bone ingrowth and osseointegration at 7, 10, 14, 28, 56, and 84 days after implantation. The results showed that microroughening of the bioactive glass surface significantly enhanced the bone-bonding response of the biomaterial. The positive response was seen in one of the three bioactive glass compositions studied. The affinity index of new bone on the glass surface was significantly (p = 0.02) increased with a trend (p = 0.10) toward improved mechanical incorporation. New bone formation was dependent on the glass composition, and it was found to occur not only through the mechanism of bone ingrowth but also based on in situ osteogenesis within implant interstices. Based on these results, the procedure of microroughening could enhance the osteopromotive properties of certain bioactive glass compositions. PMID- 14566791 TI - Biocompatibility of collagen membranes crosslinked with glutaraldehyde or diphenylphosphoryl azide: an in vitro study. AB - Crosslinking of collagen biomaterials increases their resistance to degradation in vivo. Glutaraldehyde (GA) is normally used to crosslink collagen biomaterial, but is often cytotoxic. Diphenylphosphoryl azide (DPPA) has recently been proposed as reagent, but little is known about its effects on cell behavior. In this study, we determined which collagen membrane was the most biocompatible: Paroguide which is crosslinked with DPPA and contains chondroitin sulfate; Opocrin which is crosslinked with DPPA; Biomed Extend which is crosslinked with GA; and Bio-Gide which is left untreated. Cell proliferation and extracellular matrix macromolecule deposition were evaluated in human fibroblasts cultured on the membranes. The GA-crosslinked Biomed Extend membrane and the not-crosslinked Bio-Gide membrane reduced cell growth and collagen secretion compared with DPPA crosslinked biomembranes. When Paroguide and Opocrin were compared, better results were obtained with Paroguide. The greatest amount of transforming growth factor beta1, a growth factor involved in extracellular matrix macromolecule accumulation and in tissue regeneration, was produced by cells cultured on Paroguide, with Opocrin second. Our data suggest that the DPPA method is more biocompatible than the GA for crosslinking collagen biomaterials and that membranes made of collagen plus chondroitin sulfate are better than membranes made of pure collagen. PMID- 14566792 TI - Local carboplatin delivery and tissue distribution in livers after radiofrequency ablation. AB - This study investigated the local drug pharmacokinetics of intralesional drug delivery after radiofrequency ablation of the liver. We hypothesized that the tissue architecture damaged by the ablation process facilitates the drug penetration in the liver and potentially enlarges the therapeutic margin in the local treatment of cancer. The delivery rate and tissue distribution of carboplatin, an anticancer agent, released from poly(D,L-lactide-co-glycolide) implants into rat livers after radiofrequency ablation were quantified by atomic absorption spectroscopy. Results showed that carboplatin clearance through blood perfusion was significantly slower in the ablated livers, leading to a more extensive tissue retention and distribution of the drug. The concentration of Pt at the implant-tissue interface ranged from 234 to 1440 microg Pt/(g liver) in the ablated livers over 144 h versus 56 to 177 microg Pt/(g liver) in the normal tissue. The maximum penetration distance at which Pt level reached above 6 microg/g (calculated based on a reported IC90 value for carboplatin) was 8-10 mm and 4-6 mm in ablated and normal liver, respectively. Histological analysis of the necrotic lesions showed widespread destruction of tissue structure and vasculature, supporting the initial hypothesis. This study demonstrated that intralesional drug delivery could provide a sustained, elevated concentration of anticancer drug at the ablation boundary that has the potential to eliminate residual cancer cells surviving radiofrequency ablation. PMID- 14566793 TI - Interactions of heparin with human skin cells: binding, location, and transdermal penetration. AB - The development of new materials for tissue engineering of skin substitutes requires an increasing knowledge of their interactions with human skin cells. Since carbohydrate recognition is involved in numerous biologic processes, including skin regeneration, the aim of this study was to identify sugar receptors expressed at the surface of human dermic and epidermic cells. Binding of fluorescent sugar-polyhydroxyethylacrylamide derivatives was analyzed by flow cytofluorimetry on cultured human skin fibroblasts, keratinocytes, and melanocytes. We observed that these three cell types express a membrane receptor specific for GlcNAc6S. Since the polysaccharide heparin contains this sugar moiety, we further investigated the interactions of heparin with skin cells. We analyzed the in vitro cell binding and ex vivo diffusion with the Franz cell of heparin and of two other polysaccharides of similar molecular weight, dextran and chondroitin sulfate. We found evidence of the preferential binding of heparin on keratinocytes and its high transcutaneous penetration of skin. Altogether, our results describe the affinity of heparin for human skin cells and suggest it may be an excellent candidate for use in the skin delivery of drugs or cosmetics and also as an active component in engineered skin. PMID- 14566794 TI - A study on the effect of dual blasting with TiO2 on titanium implant surfaces on functional attachment in bone. AB - In the present study, the effect of a dual treatment of titanium implants and the subsequent bone response after implantation were investigated. Coin-shaped c.p. titanium implants were placed into the tibias of 12 rabbits. The implant, which was dually blasted with TiO2 particles of two different sizes, was compared with implants that were blasted with only one of these particle sizes. Implants in group 1 were grit blasted with small particles, 22-28 microm in size, and group 2 with coarser particles, 180-220 microm size. These two treatments gave different surface micro textures. To test the effect of a combination of two different treatments, group 3 implants were blasted first with the 180- to 220-microm and subsequently with the 22- to 28-microm particles. The surface topography of the implant specimens was examined by scanning electron microscopy and by a confocal laser scanner and a numeric evaluation of S(a), S(t), and S(dr) was recorded. Group 2 implants, which were blasted with only the coarse particles, showed a significantly better functional attachment (p < 0.001) than the other two groups. Group 1, which was blasted with only small particles, showed the lowest retention in bone. There was a positive correlation between the topographical and mechanical evaluation of the surfaces. PMID- 14566795 TI - Nano-fibrous scaffolding architecture selectively enhances protein adsorption contributing to cell attachment. AB - Tissue engineering aims at resolving problems such as donor shortage and immune rejection faced by transplantation. Scaffolds (artificial extracellular matrices) have critical roles in tissue engineering. Recently, we developed nano-fibrous poly(L-lactic acid) scaffolds under the hypothesis that synthetic nano-fibrous scaffolding, mimicking the structure of natural collagen fibers, could create a more favorable microenvironment for cells. This is the first report that the nano fibrous architecture built in three-dimensional scaffolds improved the features of protein adsorption, which mediates cell interactions with scaffolds. Scaffolds with nano-fibrous pore walls adsorbed four times more serum proteins than scaffolds with solid pore walls. More interestingly, the nano-fibrous architecture selectively enhanced protein adsorption including fibronectin and vitronectin, even though both scaffolds were made from the same poly(L-lactic acid) material. Furthermore, nano-fibrous scaffolds also allowed >1.7 times of osteoblastic cell attachment than scaffolds with solid pore walls. These results demonstrate that the biomimetic nano-fibrous architecture serves as superior scaffolding for tissue engineering. PMID- 14566796 TI - Adhesive protein interactions with chitosan: consequences for valve endothelial cell growth on tissue-engineering materials. AB - Stable endothelialization of a tissue-engineered heart valve is essential for proper valve function, although adhesive characteristics of the native valve endothelial cell (VEC) have rarely been explored. This research evaluated VEC adhesive qualities and attempted to enhance VEC growth on the biopolymer chitosan, a novel tissue-engineering scaffold material with promising biological and chemical properties. Aortic VEC cultures were isolated and found to preferentially adhere to fibronectin, collagen types IV and I over laminin and osteopontin in a dose-dependent manner. Seeding of VEC onto comparison substrates revealed VEC growth and morphology to be preferential in the order: tissue culture polystyrene > gelatin, poly(DL-lactide-co-glycolide), chitosan > poly(hydroxy alkanoate). Adhesive protein precoating of chitosan did not significantly enhance VEC growth, despite equivalent protein adsorption as to polystyrene. Initial cell adhesion to protein-precoated chitosan, however, was higher than for polystyrene. Composite chitosan/collagen type IV films were investigated as an alternative to simple protein precoatings, and were shown to improve VEC growth and morphology over chitosan alone. These findings suggest potential manipulation of chitosan properties to improve amenability to valve tissue-engineering applications. PMID- 14566797 TI - Use of AFM to probe the adsorption strength and time-dependent changes of albumin on self-assembled monolayers. AB - The adsorption kinetics of human serum albumin (HSA) on CH3- and COOH-terminated self-assembled monolayers (SAMs) has been investigated using radioassays and atomic force microscopy (AFM). On both surfaces, the amount of HSA adsorbed reached a plateau after 30 min. The plateau level was higher on the CH3 compared to the COOH surface. The adhesion force (Fadh), measured using Si3N4 AFM tips in water, decreased with time of contact with the HSA solution on the CH3 surface. This time-dependent change in the adhesiveness of the adsorbed protein is best explained by a change in the conformation or orientation. In contrast, Fadh was independent of the time of contact with the HSA solution on the COOH surface, indicating that once adsorbed, the HSA molecules do not undergo further conformation or orientation changes. The perturbation induced by scanning with the AFM in water on the adsorbed HSA layers was greater on CH3 surfaces than on COOH surfaces, suggesting a weaker protein-substratum interaction on the CH3 terminated SAMs. This was further confirmed by a stronger desorption of HSA following sodium dodecyl sulfate (SDS) treatment on the CH3 surface compared to the COOH surface. Taken together, these data suggest that for COOH SAMs, (1) there is a strong interaction between HSA and the substratum; (2) there is an absence of reorientation with time; and (3) there is a smaller amount of adsorbed protein at 24 h, possibly due to increased but rapid spreading/denaturation of the protein. On the CH3 surface, less deformation of HSA occurs and the molecules maintain a higher mobility at short adsorption times. AFM measurements performed after aging of an adsorbed HSA layer in buffer suggests the role played by HSA in solution in determining the time-dependent conformation and/or orientation changes. PMID- 14566798 TI - Human bone marrow stromal cell and ligament fibroblast responses on RGD-modified silk fibers. AB - Adhesion, spreading, proliferation, and collagen matrix production of human bone marrow stromal cells (BMSCs) on an RGD-modified silk matrix was studied. Anterior cruciate ligament fibroblasts (ACLFs) were used as a control cell source. Scanning electron microscopy (SEM) and MTT analyses demonstrated that the modified silk matrices support improved BMSC and ACLF attachment and show higher cell density over 14 days in culture when compared with the non-RGD-modified matrices. Collagen type I transcript levels (at day 7) and content (at day 14) was significantly higher on the RGD-modified substrate than on the nonmodified group. The ability of RGD-coupled silk matrices to support BMSC attachment, which leads to higher cell density and collagen matrix production in vitro, combined with mechanical, fatigue, and biocompatibility properties of the silk protein matrix, suggest potential for use of this biomaterial for tissue engineering. PMID- 14566799 TI - Fatigue and fracture toughness of acrylic bone cements modified with long-chain amine activators. AB - The composition of acrylic bone cement has been identified as one of the important parameters affecting its mechanical properties and may, in turn, ultimately influence the longevity of a cemented arthroplasty. Our aim in this study was to determine the influence of change of one compositional variable, the activator, on the fatigue performance and fracture toughness of specimens of the fully cured cement. To that end, three sets of cements were prepared, containing either the conventional activator, 4-N,N dimethyl p-toluidine (DMPT), or novel ones that are tertiary amines based on long-chain fatty acids, that is, 4-N,N dimethylaminobenzyl oleate (DMAO) and 4-N,N dimethylaminobenzyl laurate (DMAL). In the fatigue tests, the specimens were subjected to tension-tension loading, and the results (number of cycles to failure, Nf) were analyzed using the linearized form of the three-parameter Weibull equation. The fracture toughness (KIc) tests were conducted with rectangular compact tension specimens. All fracture surfaces were subsequently examined with scanning electron microscopy. We found that the Weibull mean fatigue lives for specimens fabricated using the DMPT, DMAL, and DMAO containing cements were 272,823, 453,551, and 583,396 cycles, respectively. The corresponding values for KIc were 1.94 +/- 0.05, 2.06 +/- 0.09, and 2.00 +/- 0.07 MPa radical m, respectively. Statistical analyses showed that for both the DMAL- and DMAO-containing cements, the mean values of Nf were significantly higher compared to the corresponding value for the DMPT containing cement (Mann-Whitney test; alpha < 0.10). This result is attributed to the higher molecular weights of the former cements compared to the latter. The same trend was found for the mean KIc values (Mann-Whitney test; alpha < 0.05), with the trend being explained in terms of the differences seen in the crack morphologies. These results thus demonstrate that these novel amines are viable alternatives to DMPT for incorporation into acrylic bone cement formulations in the future. PMID- 14566800 TI - Structure and properties of the precipitates formed from condensed solutions of the revised simulated body fluid. AB - Precipitation experiments with aqueous solutions of the Kokubo's revised simulated body fluid (rSBF) equal to 2, 4, 8, and 12 times the ionic concentration of human blood plasma were performed. Instead of Hepes, solution pH was adjusted to the desired value of 7.40 +/- 0.02 by either bubbling of CO2 or addition of HCl. The experiments were performed in tightly closed plastic vessels kept at 37.0 +/- 0.2 degrees C for 72 h under permanent shaking. Afterward, the suspensions were filtrated, and the precipitates were collected and analyzed. The results revealed that increasing the concentration of rSBF resulted in great changes in both the structure and the chemical composition of the precipitates. Phosphate substitution for carbonate (although the amounts of calcium and magnesium remained unchanged) and crystallinity decreasing were the most important modifications found in the precipitates formed from the highly condensed solutions of rSBF. PMID- 14566801 TI - Mathematical model of static platelet adhesion on a solid surface. AB - The scheme of platelet/surface interaction and a kinetic model of platelet adhesion on a solid surface are suggested. The elaborated approach takes into account the platelet activation by the surface and accumulation of free activated cells in the bulk of the liquid phase. This effect has an especially important role in static experimental conditions. The suggested model explains three types of adhesion kinetic curves, obtained in experiments in vitro: sigmoid curves with or without saturation and an exponential curve with saturation. According to the model, the curve shape is determined by material surface properties, platelet functionality, and experimental conditions of the platelet/surface interaction. The data of static platelet adhesion from platelet rich plasma on glass, siliconized glass, hexadecyltrichlorosilane monolayers, and low-density polyethylene are described mathematically with the proposed model. Numerical parameters are calculated from approximation of experimental data by the model. These parameters allow quantitative characterization of platelet interaction with the material surface. PMID- 14566802 TI - Development of a porous poly(L-lactic acid)/hydroxyapatite/collagen scaffold as a BMP delivery system and its use in healing canine segmental bone defect. AB - A hydroxyapatite/collagen (HAC) composite was produced to mimic the natural extracellular matrix of bone, with the collagen serving as a template for apatite formation. A three-dimensional highly porous scaffold was developed by mixing HAC with poly(L-lactic acid) (PLA) using a thermally induced phase separation technique. Naturally derived bovine bone morphogenetic protein (bBMP) was incorporated into the porous HAC-PLA scaffolds, and the composite then was implanted in diaphyseal defects (2 cm in radius) of adult beagle dogs. Controls were implanted with scaffolds without BMP. The dogs were sacrificed at 6 months, at which time biocompatibility, biodegradability, and osteoinduction were evaluated by histologic and radiologic examination and by bone mineral density (BMD) measurements. All defects healed after treatment with BMP combined with HAC PLA, and BMD at the site of the defect was higher than the BMD of the intact radius. Fibrous union developed in the control group animals. Histologic observation indicated that the presence of BMP not only promoted osteogenesis but that it also accelerated degradation of the biomaterials. Optimized design parameters of a three-dimensional porous biomaterial would give full scope to the role of BMP as an osteoinductive growth factor. PMID- 14566803 TI - Ca/P ratio effects on the degradation of hydroxyapatite in vitro. AB - Phase purity is a well-recognized but not well-understood variable affecting the biological integration of hydroxyapatite (HA)-based biomaterials. Minor amounts of specific, relevant impurities--calcium oxide (CaO) and tricalcium phosphate (TCP)--may often be present either as deliberate additions or as a result of decomposition during sintering. We investigated the influence of these two impurities in terms of their effects on surface morphology, weight loss/gain, and microstructural-level degradation. Phase purity variations were deliberately introduced into an otherwise-standardized HA matrix--the parent HA grain size and bulk density were relatively constant--produced using identical fabrication conditions. Stability varied markedly during exposure to mildly acidic, neutral, and pH 7.4 phosphate-buffered saline. Equivalent molar variations in the Ca/P ratio (1.62 vs 1.72) on either side of the stoichiometric ratio produce relatively small volumetric amounts of CaO (1.6 vol%) versus TCP (27 vol%) in HA. However, the relatively small amounts of CaO render the bulk more susceptible to degradation and more likely to have negative effects on a biological milieu. Interestingly, the presence of CaO is also a potent nucleating agent for the precipitation of new surface phases and detectable weight gain. The TCP containing ceramic, in contrast, paradoxically exhibited slightly greater resistance to degradation than HA. PMID- 14566805 TI - Biologically inspired synthesis of bone-like composite: self-assembled collagen fibers/hydroxyapatite nanocrystals. AB - Replacement of bone tissue by graft materials and products of tissue engineering having composition, structure, and biological features that mimic natural tissue is a goal to be pursued. A biomimetic synthesis was performed to prepare new bone like composites constituted of hydroxyapatite nanocrystals and self-assembled type I collagen fibers. We used a biological inspired approach that proved that the biological systems stored and processed information at the molecular level. Two different methodologies were used: dispersion of synthetic hydroxyapatite in telopeptides free collagen molecules solution and direct nucleation of hydroxyapatite into reconstituted collagen fibers during their assembling. The different preparation techniques were experimented then the composites thoroughly characterized and compared. Composite obtained by direct nucleation showed an intimated interaction of the inorganic and proteic components, which modified the apatitic phase and made its composition, morphology and structure similar to the mineral component of natural bone. PMID- 14566804 TI - Novel complexation hydrogels for oral peptide delivery: in vitro evaluation of their cytocompatibility and insulin-transport enhancing effects using Caco-2 cell monolayers. AB - Poly[methacrylic acid-grafted-poly(ethylene glycol)] [P(MAA-g-EG)] is a complexation hydrogel molecularly designed for oral peptide delivery. In this work, the cytotoxicity and insulin-transport enhancing effect of P(MAA-g-EG) microparticles on intestinal epithelial cells were evaluated using Caco-2 cell monolayers. A series of P(MAA-g-EG) microparticles with different polymer compositions were prepared by a photo-initiated free radical solution polymerization and subsequent pulverization. The hydrogel microparticles were preswollen in either Ca2+-containing (CM+) or Ca2+-free medium (CM-; pH 7.4) and applied to the apical side of the Caco-2 monolayers. No significant cytotoxic effects, as determined by a calorimetric assay with P(MAA-g-EG) microparticles preswollen in the CM+, were observed at doses ranging from 3 to 31 mg/cm2 of cell monolayer. Transepithelial electrical resistance (TEER) measurements showed that the P(MAA-g-EG) microparticles induced a Ca2+ concentration-dependent lowering in TEER values. The reduction effect in CM- media was greater than that in CM+ media (17 +/- 2% reduction in CM+ and 45 +/- 3% reduction in CM-, respectively). Insulin transport in the presence of the preswollen P(MAA-g-EG) microparticles was also strongly depended on the Ca2+ concentration in the medium. The respective estimated permeability for insulin alone and the insulin with hydrogels in CM+ were 0.77 and 1.16 x 10(-8) cm/s, whereas those in CM- were 1.18 and 24.78 x 10(-8) cm/s. The results demonstrate that the P(MAA-g-EG) hydrogel microparticles could be used as a cytocompatible carrier possessing the transport enhancing effect of insulin on the intestinal epithelial cells. PMID- 14566806 TI - Immunocamouflage: prevention of transfusion-induced graft-versus-host disease via polymer grafting of donor cells. AB - Graft-versus-host disease (GVHD) can occur following the transfer of allogeneic lymphocytes into immunosuppressed and, in rare cases, immunocompetent recipients. The initiation of GVHD requires the allorecognition of the recipient's disparate MHC molecules by the donor T lymphocytes (T cell). Currently, GVHD is controlled by cyclosporine administration--a potent, but toxic, T-cell suppressing agent. To determine if the nontoxic grafting of methoxypoly(ethylene glycol) (mPEG) to immunologically foreign lymphocytes could prevent allorecognition and GVHD, in vitro and in vivo murine studies were performed. In vitro studies utilizing mixed lymphocyte reactions (MLRs) demonstrate that mPEG modification effectively prevented allorecognition and subsequent T-cell proliferation. The loss of cellular proliferation was not due to mPEG cytotoxicity but rather to the inhibition of cell-cell interactions. Flow cytometric studies showed that T-cell and antigen-presenting cell adhesion molecules (CD2, CD11a), signaling (CD3epsilon, T-cell receptor), and costimulatory molecules (CD28, CD80) were efficiently immunocamouflaged by mPEG derivatization. Interestingly, upon antigenic stimulation mPEG-modified cells demonstrate enhanced apoptosis as evidenced by DNA laddering. In vivo studies using immunocompetent and immunosuppressed mice established that mPEG modification of donor lymphocytes effectively attenuated the in vivo proliferation of donor cells and the initiation of GVHD. PMID- 14566807 TI - Transforming growth factor-beta1 in a sterilized tissue derived from the pig small intestine submucosa. AB - The extracellular matrices of connective tissues contain growth factors such as transforming growth factor (TGF)-beta1. The possibility arises, therefore, that animal connective tissues that have been excised and rendered acellular in the sterilization, lyophilization, and other preparative processes for human use may still retain active growth factors that could contribute to the clinical efficacy of the product. We therefore analyzed 4M guanidine HCl extracts of a sterilized, acellular matrix, Oasis Wound Matrix, for the presence of TGF-beta1 by a sandwich enzyme-linked immunosorbent assay using the soluble type II receptor for TGF-beta to capture the growth factor, and for biological activity by testing the capacity of the extracts to inhibit 3[H]thymidine incorporation into Mv1Lu cells (mink lung epithelial cells). The enzyme-linked immunosorbent assay determined that TGF beta1 was present at a concentration of 710.9 +/- 157.7 pg/g dry weight of tissue in Oasis and 768.1 +/- 182.1 pg/g dry weight of tissue in SIS (porcine small intestinal submucosa), the disinfected precursor of Oasis. The growth inhibition assays demonstrated that the Oasis extracts inhibited the proliferation of Mv1Lu cells in culture, consistent with the TGF-beta1 in the material having biological activity. Most of the TGF-beta1 survives the sterilization and lyophilization processes in the preparation of the Oasis Wound Matrix, and is functional in its ability to bind to its receptor and, apparently, in its capacity to inhibit growth. PMID- 14566808 TI - In vitro cytotoxicity evaluation of a 50.8% NiTi single crystal. AB - To our knowledge, the biocompatibility of nickel-titanium (NiTi) single crystals has not been reported. Yet certain orientations of single crystals present several advantages over the polycrystalline form in terms of maximal strain, fatigue resistance, and temperature range of superelasticity. Therefore we tested the in vitro biocompatibility of 50.8% NiTi single crystals in the orientation <001> after four different heat treatments in a helium atmosphere followed by mechanical polishing. The study was performed on the material extracts after immersion of the specimens in cell culture medium (DMEM) for 7 days at 37 degrees C. Cytotoxicity studies were performed on L-929 mouse fibroblasts using the MTT assay. J-774 macrophages were used to assess the potential inflammatory effect of the extracts by IL1-beta and TNF-alpha dosages (sandwich ELISA method). Exposure of L-929 to material extracts did not affect cell viability. In addition, IL1 beta and TNF-alpha secretion was not stimulated after incubation with NiTi extracts compared to the negative controls. These results were predictable since atomic absorption spectroscopy did not detect nickel ions in the extracts with a resolution of 1 ppm. Within the limits of in vitro testing, our results demonstrate that the TiNi(50.8%) single crystals do not trigger a cytotoxic reaction. PMID- 14566809 TI - Fibronectin anchorage to polymer substrates controls the initial phase of endothelial cell adhesion. AB - Early stages of the adhesion of human endothelial cells onto a set of smooth polymer films were analyzed to reveal the modulation of cell-matrix interactions by the physicochemical constraints of predeposited fibronectin (FN). Hydrophobic and hydrophilic polymer substrates, consisting of poly(octadecene-alt-maleic anhydride) and poly(propene-alt-maleic anhydride) films, were coated with similar amounts of FN at conditions of either covalent or noncovalent immobilization. The well-defined substrates permit variation of the anchorage of FN at invariant topography, pliability, and molecular composition. Although all of the compared FN coatings were effective in stimulating attachment of endothelial cells, the initial formation of cell-matrix adhesions was found to be controlled by the type of interaction between predeposited FN and the underlying substrate. Covalent linkage and hydrophobic interactions of the predeposited FN with the polymer films interfered with the rapid generation of focal and fibrillar adhesions. It was demonstrated that this was caused by the fact that only weakly bound FN could become readily reorganized by the adherent cells. Upon prolonged culture periods at standard cell culture conditions, secretion and deposition of organized extracellular matrix by the attached cells was found to balance out the differences of the substrates. PMID- 14566810 TI - Examination of membrane rupture as a mechanism for mammalian cell detachment from fibronectin-coated biomaterials. AB - Synthetic biomaterials intended for the reconstruction of tissues and organs must be capable of sustaining adhesive contact with adjacent cells and tissues under mechanical and hydrodynamic stresses. To facilitate this adhesion, extracellular matrix proteins or peptide sequences are frequently immobilized to the biomaterial interface. These ligands enhance cell attachment by raising the number of cell receptor/ligand interactions, but consequently they may alter the mechanism of cell detachment. In particular, as the cell membrane is more strongly immobilized to the substratum, the tendency for cell detachment to involve membrane rupture may increase. To test this hypothesis, cells were fluorescent stained with a membrane dye, allowed to attach to fibronectin-coated model substrates for 30 min, and then subjected to a spatially dependent range of shear stress for 5 min (28-220 dyn/cm2) using a radial-flow chamber. Phase contrast and fluorescent images were analyzed to determine the probability for cell detachment and the area of fluorescent debris left by detaching cells as a function of fibronectin concentration, magnitude of shear stress, and time. It was found at all concentrations of fibronectin that the majority of detaching cells left membrane fragments, the mean size of these fragments was independent of shear stress, and the shape independent of the direction of flow. However, mean fragment area increased with concentration of fibronectin and decreased with duration of shearing flow. We postulate that the area of debris reflects the extent of cell attachment prior to the application of shear and that adhesive complexes can disassemble at the onset of flow. PMID- 14566811 TI - Extended delivery of the antimitotic agent colchicine from thermoresponsive N isopropylacrylamide-based copolymer films to human vascular smooth muscle cells. AB - The aim of this study was to establish the capacity of thermoresponsive poly(N isopropylacrylamide) copolymer films to deliver bioactive concentrations of an antimitotic agent to human vascular smooth muscle cells (HASMC) over an extended period of time. Copolymer films were prepared using a 50:50 (w/w) ratio of N isopropylacrylamide (NiPAAm) monomer to the more hydrophobic N-tert butylacrylamide (NtBAAm) and loaded with the antimitotic agent colchicine (0.1 micromol per film) at room temperature. Colchicine release from films was sustained over a 14-day period. At 24 h postloading, the concentration of colchicine in the medium overlying films was 2.12 +/- 0.16 microM; this fell to 0.20 +/- 0.01 microM at 7 days and decreased further to 0.12 +/- 0.01 microM after 14 days. Colchicine released from copolymer films inhibited proliferation when subsequently placed on HASMC: at 0.1 microM, released colchicine reduced proliferation to 18.5 +/- 0.8% of control cells (p < 0.001, n = 9). The antiproliferative effect of released colchicine was comparable to that of native colchicine, as observed in separate experiments. Furthermore, colchicine released from 50:50 polymer films inhibited the proliferation of cells grown in the same environment as the copolymer. Inhibition of cell proliferation was not due to the release of cytotoxic particles from the copolymer because medium incubated with copolymer for 5 days and then applied to HASMC did not alter cell viability. In conclusion, this study demonstrates that 50:50 NiPAAm:NtBAAm copolymers can deliver bioactive concentrations of the antimitotic agent colchicine to human vascular cells over an extended period of time. PMID- 14566812 TI - Localized corrosion of 316L stainless steel with SiO2-CaO films obtained by means of sol-gel treatment. AB - Sol-gel films on austenitic stainless steel (AISI 316L) polished wafer were prepared from sono-sols obtained from tetraethylorthosilane and hydrated calcium nitrate. However, pitting was observed in different places on the stainless steel surfaces. The corrosion resistance was evaluated by the polarization resistance in simulated body fluid environment at 37 degrees C. The critical current density, the passive current density, the corrosion potential, and the critical pitting potential were studied. The austenitic stainless steel 316L treated presents important electrochemical corrosion and consequently its application as endosseous implants is not possible. PMID- 14566813 TI - Effective removal of alginate-poly-L-lysine microcapsules from pancreatic islets by use of trypsin-EDTA. AB - Although the transplantation of alginate-poly-L-lysine-alginate encapsulated islets of Langerhans usually is successful, graft survival is still limited. Molecular analysis by RT-PCR of the encapsulated islets may provide insight into the mechanisms that affect islets during graft failure. However, RT-PCR on encapsulated islets is not possible because the poly-L-lysine of the capsule interferes with both cDNA synthesis and PCR amplification. We applied a method that mechanically removes the microcapsules from the islets after a short trypsin EDTA treatment (decapsulation), thereby enabling RT-PCR analysis. The results of this study show that the decapsulation procedure does not affect islet vitality and has only minor effects on islet function and morphology. The decapsulation does not affect GAPDH, beta-actin, Bcl-2, or Bax gene expression. This method is an improvement over the time-consuming manual dissection of microcapsules because it allows for the rapid and relatively harmless removal of capsules on a larger scale. Decapsulation offers the possibility of applying RT-PCR, as well as other methods, which cannot be performed on encapsulated islets. PMID- 14566814 TI - Direct attachment of fibronectin to tresyl chloride-activated titanium. AB - The aim of this study was to bind fibronectin directly to a titanium surface treated with tresyl chloride (2,2,2-trifluoroethanesulfonyl chloride) for the development of a strong connection of a dental implant to subepithelial connective tissues and/or peri-implant epithelia. Basic terminal OH groups of mirror polished titanium were allowed to react with tresyl chloride at 37 degrees C for 2 days. The tresylated titanium disk was then immersed into a fibronectin/phosphate-buffered saline solution for 24 h at 37 degrees C. The activation reaction of the basic OH of titanium with tresyl chloride was confirmed by S2p, F1s, and O1s spectra using X-ray photoelectron spectroscopy and -O-S-O2- bonds using Fourier transform infrared reflection-absorption spectroscopy. After the reaction of fibronectin with titanium, the X-ray photoelectron spectroscopy revealed the remarkable effect of the activation of terminal OH groups with the tresyl chloride treatment. The N1s peak derived from the attached fibronectin still remained after 60 s of argon-ion sputtering after tresyl chloride treatment. In contrast, the N1s peak of the specimen not treated with tresyl chloride almost disappeared after only 10 s of argon-ion etching. Fibronectin, a well-known cell-adhesive protein, could easily be attached to the titanium surface by use of the tresyl chloride activation technique. PMID- 14566815 TI - SV40 in human cancers--an endless tale? PMID- 14566817 TI - Detection of point mutations of the Axin1 gene in colorectal cancers. AB - Axin is a recently identified tumor suppressor that plays an important role in liver and colon cancers. To gain further insights into the structure and function of Axin in controlling cell growth, we analyzed 54 colorectal cancer tissues for mutations in AXIN1 gene. We employed PCR amplification with 23 sets of primers against introns that encompassed the whole coding region of AXIN1 followed by single-strand conformation polymorphism (SSCP) analysis. After subcloning and sequencing analysis of the reamplified DNA from the aberrant bands, we found, in addition to 3 silent mutations, 6 missense point mutations in different functionally important regions. The missense mutation rate is hence 11%, suggesting that Axin deficiency may contribute to the onset of colorectal tumorigenesis. PMID- 14566816 TI - Tumor reversion: correction of malignant behavior by microenvironmental cues. AB - Cancer is characterized by unrestrained proliferation and loss of organization, a process that is intimately linked to, and controlled by, reciprocal signaling between the genetically altered tumor epithelium, the stroma, the components of the basement membrane and inflammatory mediators. Much work has been done to characterize the genetics of cancer cells. In this review, we describe the experiments that have been performed, which point to the significant role of the tissue microenvironment in the developmental regulation of normal and neoplastic cells. Using a variety of model systems, the works of a number of laboratories have converged on a hypothesis where the correction of 1 or 2 signaling defects can revert tumor cells to a normal phenotype, both in vivo and in culture, even when the tumor cells possess multiple genetic and epigenetic lesions. This paradigm has been successfully used to treat acute promyelocytic leukemia, and it remains the task of biomedical researchers to identify additional targets for the reversion of other human malignancies. PMID- 14566818 TI - Reporter gene assay demonstrates functional differences in estrogen receptor activity in purified breast cancer cells: a pilot study. AB - Tamoxifen has contributed to a dramatic reduction in breast cancer mortality and recent results indicate that aromatase inhibitors may further improve survival in some patients. Nevertheless, a substantial proportion of patients become resistant to treatment. To date, with the exception of estrogen receptor (ER) determination by ligand binding or immunohistochemical techniques, there has been no way of predicting which of several therapies is indicated in particular patients. We describe a novel assay using the adenoviral gene delivery system to assess ER function in breast cancer cells derived directly from patients. The purification and short-term culture of these cells has been recently described by our laboratory. Adenovirus containing an estrogen-regulated beta-galactosidase reporter gene (ERE-lacZ) was constructed and used to test ER activity in breast cancer cells derived from 18 patients with primary and 16 patients with metastatic cancer, under varying treatment schedules. The adenoviral assay enabled ER activity to be readily determined in purified cells from primary breast cancers and secondary sites. Breast cancers cells could be categorized on the basis of ER activity in the absence of ligand, the presence of estrogen or anti-estrogens. In primary breast cancers, our results correlated with ER determination by immunohistochemistry in 78% of cases. In patients who had become resistant to tamoxifen, however, we found some in whom reporter activity was stimulated by tamoxifen and others whose tumors were either still estrogen responsive or completely unresponsive, irrespective of the original ER content. Our findings indicate that this reporter assay could be useful in decisions regarding use of adjuvant endocrine therapies in breast cancer. PMID- 14566819 TI - Autocrine motility factor signaling induces tumor apoptotic resistance by regulations Apaf-1 and Caspase-9 apoptosome expression. AB - Autocrine motility factor (AMF) is a cytokine that regulates locomotion and metastasis of tumor cells. It is well known that expression levels of AMF secretion and its receptor (AMF R) are closely related to tumor malignancy and rheumatoid arthritis. We have established that AMF signaling induced anti apoptotic activity and that human fibrosarcoma HT-1080 line that secreted high levels of AMF were resistant to drug-induced apoptosis. These cells did not express the apoptotic protease activating factor-1 (Apaf-1) and Caspase-9 genes that encode for the proteins that form the "apoptosome" complex. The disappearance of the Apaf-1 and Caspase-9 gene was recovered by a cellular signaling inhibitor of protein kinase C, phosphatidylinositol 3-phosphate kinase and mitogen-activated protein kinase of the in vitro cultured human fibrosarcoma HT-1080 line. Treatment with these inhibitors favored apoptotic cell death induced by anti-cancer drugs of the murine ascites Ehrlich line. Apoptotic resistance of tumor cells allows them to escape death from cancer chemotherapy, so an understanding of malignant anti-apoptotic activities is important. Antibodies against AMF induced Ehrlich ascites apoptosis in vitro, and effectively aided in vivo apoptosis induced by anti-cancer drugs. The results might indicate a novel route by which tumor cells protect themselves with products, such as AMF, and proliferate despite various stresses and chemical insults; AMF regulates expression of Apaf-1 and caspase-9 genes via a complex signaling pathway and indirectly regulates formation of the apoptosome. PMID- 14566820 TI - Cyclic AMP inhibits TGFbeta1-induced cell-scattering and invasiveness in murine transformed keratinocytes. AB - Mouse-transformed keratinocytes cultured in the presence of transforming growth factor beta1 (TGF-beta1) acquire an array of morphologic and functional properties that give rise to a migratory phenotype that expresses mesenchymal molecular markers. This cellular conversion involves activation of the Ras-ERK pathway, enhancement of urokinase (uPA) and matrix metalloproteinase-9 (MMP-9) expression and induction of invasiveness. In our present work, we demonstrate that cAMP and forskolin are able to prevent the expression of these mesenchymal properties, probably due to blockade of the Ras-ERK pathway. Our results also show that cAMP and forskolin are able to abolish the TGF-beta1-induced reorganization of the actin cytoskeleton that is characteristic of the mesenchymal phenotype and also inhibits the disruption of the E-cadherin cell to cell interactions. The latter responses seem to depend on the activity of protein kinase A, as demonstrated by the activation of the Ras-ERK pathway by specific protein kinase A inhibitors. PMID- 14566821 TI - Reversal of breast cancer resistance protein-mediated drug resistance by tryprostatin A. AB - MDR in human cancers is one of the major causes of failure of chemotherapy. A member of the superfamily of ABC transporters, BCRP, was demonstrated to confer an atypical MDR phenotype to tumor cells. To overcome the BCRP-mediated drug resistance, the fungal secondary metabolite TPS-A, a diketopiperazine, was analyzed with regard to its potency to reverse the BCRP-mediated drug-resistant phenotype. At concentrations of 10-50 microM, TPS-A reversed a mitoxantrone resistant phenotype and inhibited the cellular BCRP-dependent mitoxantrone accumulation in the human gastric carcinoma cell line EPG85-257RNOV, the human breast cancer cell line MCF7/AdrVp (both exhibiting acquired BCRP-mediated MDR) and the BCRP cDNA-transfected breast cancer cell line MCF-7/BCRP clone 8. No cytotoxicity was seen at effective concentrations. These data indicate that TPS-A is a novel BCRP inhibitor. PMID- 14566822 TI - The effect of nitric oxide on cyclooxygenase-2 (COX-2) overexpression in head and neck cancer cell lines. AB - The overexpression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) has been previously reported in head and neck squamous cell carcinoma (HNSCC), as well as in many cancers. We hypothesized that endogenous nitric oxide (NO) might increase the expression of COX-2 in cancer cells. Therefore, we investigated the cross-talk between NO and the prostaglandin (PG) pathways in HNSCC cell lines. We found that COX-2 and iNOS expressions were elevated simultaneously. On adding the NO donor, SNAP, the PGE2 level was increased 2-20 times due to increased COX-2 expression. This increase of COX-2 expression by SNAP or PMA (potent inducer of both iNOS and COX-2) was blocked to various degrees by NO scavengers and NOS inhibitors (L-NAME and 1400W). Also, the expression of COX-2 in resting cells was inhibited by NOS inhibitors. Moreover, COX-2 expression, induced by SNAP, was inhibited by ODQ, a soluble guanylate cyclase (sGC) inhibitor. The effect of dibutyryl-cGMP on COX-2 expression was similar to that of SNAP. These results imply that endogenous or exogenous NO activates sGC and that the resulting increase of cGMP induces a signaling that upregulates the expression of COX-2 in HNSCC cell lines. We also observed that NO increased COX-2 expression in different cancer cell lines, including cervic and gastric cancer cell lines. These findings further support the notion that NO can be associated with carcinogenesis through the upregulation of COX-2, and that NOS inhibitor may be also useful for cancer prevention. PMID- 14566823 TI - Hyaluronan synthase-3 is upregulated in metastatic colon carcinoma cells and manipulation of expression alters matrix retention and cellular growth. AB - HA is a glycosaminoglycan that is synthesized on the inner surface of the plasma membrane and secreted into the pericellular matrix. HA and its biosynthetic enzymes (HAS1, HAS2 and HAS3) are thought to participate in tumor growth and cancer progression. In our study, colon carcinoma cells isolated from a lymph node metastasis (SW620) produced more pericellular HA and expressed higher levels of HAS3 mRNA compared to cells isolated from a primary colon carcinoma (SW480). To assess functionality, HAS3 expression in SW620 cells was inhibited by transfection with an asHAS3 construct. Decreased HA secretion and cell-surface retention by asHAS3 transfectants were confirmed using competitive binding and particle exclusion assays. Anchorage-independent growth, a correlate of tumor growth in vivo, was assessed by colony formation in soft agar. SW620 cells stably transfected with asHAS3 demonstrated significant growth inhibition, as evidenced by fewer colonies and smaller colony area than either SW620 cells or cells transfected with vector alone. Addition of exogenous HA restored growth in asHAS3 transfectants. Thus, we demonstrate that pericellular HA secretion and retention and HAS3 expression are increased in metastatic colon carcinoma cells relative to cells derived from a primary tumor. Inhibition of HAS3 expression in these cells decreased the pericellular HA matrix and inhibited anchorage-independent growth. These data suggest that HA and HAS3 function in the growth and progression of colon carcinoma. PMID- 14566824 TI - Freshly isolated bone marrow cells induce death of various carcinoma cell lines. AB - In some carcinomas such as digestive tract carcinomas, bone marrow infiltration by tumor cells is a frequent event but usually remains a micrometastatic disease and rarely induces overt bone lesions. The mechanisms responsible for the control of these metastases in the bone marrow remain poorly known. We show that freshly isolated bone marrow cells from human, murine and rat origin rapidly kill a wide range of syngeneic or xenogeneic carcinoma cell lines in culture. Further analysis of this cytotoxic process in the rat indicated that neither resident bone marrow macrophages nor NK cells were responsible for this cytotoxic effect that was restricted to a subpopulation of bone marrow cells expressing CD90 (Thy 1), a marker of hemopoietic precursors. The tumoricidal activity of these cells did not require long-term culture nor addition of exogenous cytokines or growth factors. A subset of CD90+ cells that rapidly differentiates into CD163(ED2) expressing macrophages was observed to be responsible for tumor cell killing. These macrophages induced a non-apoptotic death of tumor cells, a process that required both a direct interaction with the tumor cell and nitric oxide (NO) production through the activation of inducible nitric oxide-synthase (iNOS). This ability of pluripotent hemopoietic stem cells to rapidly differentiate into macrophages capable of killing invasive tumor cells may account for the limited expansion of micrometastases of some carcinomas in the bone marrow. PMID- 14566825 TI - Single amino acid substitutions in the transmembrane domains of breast cancer resistance protein (BCRP) alter cross resistance patterns in transfectants. AB - Breast cancer resistance protein (BCRP) is a member of ATP-binding cassette transporters that has an N-terminal ATP binding domain and a C-terminal transmembrane domain (TM). Expression of wild-type BCRP confers resistance to multiple chemotherapeutic agents such as mitoxantrone, SN-38 and topotecan, but not to doxorubicin. We made 32 BCRP mutants with an amino acid substitution in the TMs (7 E446-mutants in TM2, 15 R482-mutants in TM3, 4 N557-mutants in TM5 and 6 H630-mutants in TM6) and examined the effect of the substitutions on cellular drug resistance. PA317 cells transfected with any one of the 7 E446-mutant BCRP cDNAs did not show drug resistance. Cells transfected with any one of the 13 R482X2-BCRP cDNAs (X2 = N, C, M, S, T, V, A, G, E, W, D, Q and H, but not Y and K) showed higher resistance to mitoxantrone and doxorubicin than the wild-type BCRP-transfected cells. Cells transfected with N557D-BCRP cDNA showed similar resistance to mitoxantrone but lower resistance to SN-38 than the wild-type BCRP transfected cells. Cells transfected with N557E-, H630E- or H630L-BCRP cDNA showed similar degrees of resistance to mitoxantrone and SN-38. Estrone and fumitremorgin C reversed the drug resistance of cells transfected with R482-, N557- or H630-mutant BCRP cDNA. Cells transfected with R482G- or R482S-BCRP cDNA showed less intracellular accumulation of [3H]mitoxantrone than the wild-type BCRP-transfected cells. These results suggest that E446 in TM2, R482 in TM3, N557 in TM5 and H630 in TM6 play important roles in drug recognition of BCRP. PMID- 14566826 TI - HER-2 and TOP2A coamplification in urinary bladder cancer. AB - HER-2/NEU is one of the most frequently amplified oncogenes and a potential therapeutic target in bladder cancer. In breast cancer, the adjacent TOP2A gene, the molecular target for several anticancer drugs, is frequently coamplified together with HER-2. To study the amplification and expression of TOP2A and HER-2 and associations with tumor phenotype and clinical outcome in bladder cancer, a tissue microarray containing 2,317 bladder tumor samples was analyzed by FISH and immunohistochemistry. Overall amplification frequencies were 6.3% for HER-2 and 1.5% for TOP2A. Amplifications were most frequently seen in advanced-stage (pT2 4) tumors (HER-2 13.8%, TOP2A 3.4%). Of HER-2-amplified tumors, 56% also had alterations of TOP2A, including 14.7% coamplifications, 33.3% gains and 8% deletions. Only 17.6% of TOP2A amplifications occurred independently of HER-2 alterations. Both HER-2 and TOP2A amplifications were significantly associated with advanced tumor stage (HER-2 p < 0.0001, TOP2A p = 0.0218), high grade (p < 0.0001 for both) and protein overexpression (p < 0.0001 for both). TOP2A amplification and overexpression were linked to shortened survival in muscle invasive tumors (p = 0.0042 and 0.0077, respectively). In summary, our data suggest that HER-2 amplifications are frequently linked to alterations of the TOP2A gene in bladder cancer. The anatomy of the 17q12-q21 amplicon may be important for response to therapies targeting HER-2 or TOP2A. PMID- 14566827 TI - Orthotopic treatment model of prostate cancer and metastasis in the immunocompetent mouse: efficacy of flt3 ligand immunotherapy. AB - We established an orthotopic treatment model of prostate cancer to generate reproducible primary and metastatic carcinoma in immunocompetent C57BL/6 mice. Using an in vivo selection scheme of intraprostatic implantation of TRAMP-C1 cells, primary prostate tumors were cultured and recycled three times by intraprostatic injection resulting in the selection and establishment of the recycled cell line TRAMP-C1P3. Prostate tumors were detected approximately 30 days post-implantation with periaortic lymph node metastasis in 19/20 (95%) of mice. Tissue culture amplification, DNA ploidy and PCR amplification of the SV40 transgene were used to detect metastatic TRAMP-C1P3 in lymph node specimens. Tissue culture amplification and DNA ploidy were as sensitive as SV40 transgene amplification by PCR in detection of early metastatic disease in draining lymph nodes. To establish the use of the orthotopic model of prostate cancer for immunotherapy, mice were injected orthotopically with TRAMP-C1P3 cells and 7 days post-implantation treated daily for 28 days with either flt3L or carrier control. Carrier-treated mice had clinically detectable prostate tumors, lymph node metastasis and were moribund at 29-35 days, whereas flt3L therapy markedly suppressed primary TRAMP-C1P3 growth and lymph node metastasis, and prolonged survival. In summary, we have established a reproducible and clinically relevant orthotopic treatment model of prostate cancer in immunocompetent mice with application to a variety of therapeutic strategies. We demonstrate that flt3L treatment suppressed orthotopic prostate tumor growth and lymph node metastasis reinforcing a role for flt3L as an immunotherapeutic strategy for prostate cancer. PMID- 14566829 TI - Long-term risk of second malignant neoplasms after neuroblastoma in childhood: role of treatment. AB - The aim of our study was to quantify the risk of second malignant neoplasms (SMNs) among long-term survivors of neuroblastoma and to study the influence of treatment on this risk. We studied data from 544 5-year survival patients diagnosed with neuroblastoma before age 16 years at 8 French and British treatment centres from 1948 to 1986. After an average follow-up of 15 years (range, 5-38 years), 12 children developed a total of 13 SMNs, whereas 1.19 were expected from general population rates. Among these SMNs, there were 5 thyroid and 3 breast cancers. Increases of the risks of SMN were observed with time since neuroblastoma diagnosis and attained age. In a multivariate analysis, the relative risk of SMN associated with radiotherapy was 4.3 (95% CI 0.8-78), whereas no increased risk of SMN was associated with the administration of chemotherapy as a whole (RR = 0.4, 95% CI 0.1-1.9). Young children treated for a neuroblastoma have significantly increased risks of SMN over 3 decades of follow up. Radiotherapy treatment was found to be an important risk factor for developing SMNs, whereas no effect of chemotherapy was evidenced. Although our findings reflect the late effects of past therapeutic modalities, they underscore the importance of long-term surveillance of young children treated for a neuroblastoma. For these patients, many more years of follow-up are required to appreciate their overall risks of treatment-related SMNs. PMID- 14566828 TI - Clinical value of p53, c-erbB-2, CEA and CA125 regarding relapse, metastasis and death in resectable non-small cell lung cancer. AB - The prognostic value of p53 and c-erbB-2 immunostaining and preoperative serum levels of CEA and CA125 was investigated in a prospective multicentric study including 465 consecutive non-small cell lung cancer (NSCLC) patients with resectable tumors. Four end-points were used: lung cancer death, first relapse (either locoregional or metastasis), loco-regional recurrence and metastasis development. Standard statistical survival methods (Kaplan-Meier and Cox regression) were used. The specificity of the prognostic effect across different types of tumors was also explored, as had been planned in advance. Our results showed, once again, that pathological T and N classifications continue to be the strongest predictors regarding either relapse or mortality. Three of the studied markers seemed to add further useful information, however, but in a more specific context. For example, increased CEA concentration defined a higher risk population among adenocarcinomas but not among people with squamous tumors; and p53 overexpression implied a worse prognosis mainly in patients with well differentiated tumors. The analysis of type of relapse proved to be very informative. Thus, CA125 level was associated with a worse prognosis mainly related with metastasis development. Another interesting result was the influence of smoking, which showed a clear dose-response relationship with the probability of metastasis. For future studies, we recommend the inclusion of different endpoints, namely considering the relationship of markers with the type of relapse involved in lung-cancer recurrence. They can add useful information regarding the complex nature of prognosis. PMID- 14566830 TI - Relative risk of prostate cancer for men with affected relatives: systematic review and meta-analysis. AB - An increased risk of prostate cancer associated with a family history of prostate cancer has been documented in multiple published reports. Risk has been shown to vary by degree of relationship and age of onset of disease in the affected relative. Several studies, using various designs, have estimated the relative risk (RR) for these associations. The purpose of our study was to identify and summarize published reports on the relationship between risk of prostate cancer and family history, which is defined as having a father, brother, any first- or second-degree relative or other relative affected with prostate cancer. A Medline and manual search from 1982 to 2000 identified 24 studies that reported RR and confidence intervals (CI) and satisfied inclusion criteria. Pooled RR estimates based upon a weighted average model were as follows: any affected family member RR = 1.93, CI 1.65-2.26; affected first-degree relative RR = 2.22, CI 2.06-2.40; affected second-degree relative RR = 1.88, CI 1.54-2.30; father with prostate cancer RR = 2.12, CI 1.82-2.51; and brother with prostate cancer RR = 2.87, CI 2.21-3.73). Statistical comparison of pooled data demonstrated that the RR is significantly higher for affected brother than for affected father (p < 0.03). A sensitivity analysis demonstrated that these results are robust with respect to population bias. This meta-analysis confirms that risk of prostate cancer is associated with family history of disease and improves the quantification of this risk. PMID- 14566831 TI - Condom use promotes regression of human papillomavirus-associated penile lesions in male sexual partners of women with cervical intraepithelial neoplasia. AB - Penile HPV-associated lesions are frequently seen in male sexual partners of women with CIN. The natural course and clinical significance of these lesions are unclear. Women with CIN and their male sexual partners were randomized for condom use (condom group n = 68, noncondom group n = 68). Males were screened for the presence of penile lesions, i.e., flat lesions, papular lesions and condylomata acuminata, and of HPV in their penile swabs by PCR testing. Median follow-up time was 13.1 months (range 2.9-57.4). The outcome of our study was clinical regression of penile lesions defined as disappearance of lesions at penoscopy. Potentially prognostic factors, i.e., HPV status, lesion type and age, were studied as well. Outcomes were assessed in 57 men of the condom group and in 43 men of the noncondom group. Condom use shortened the median time to regression of flat penile lesions (7.4 months condom group vs. 13.9 months noncondom group; HR = 2.1, 95% CI 1.2-3.7). This effect was not found for papular lesions (HR = 0.5, 95% CI 0.1-2.8). HPV-negative men showed a significantly shorter median time to regression of flat lesions (3.8 months) compared to men with either HPV-positive status (8.5 months; HR = 0.4, 95% CI 0.2-0.9) or inconsistent HPV status (13.1 months; HR = 0.2, 95% CI 0.1-0.6). Regression of flat penile lesions is HPV dependent and accelerated by condom use. This effect is probably the result of blocking viral transmission between sexual partners. PMID- 14566832 TI - Condom use promotes regression of cervical intraepithelial neoplasia and clearance of human papillomavirus: a randomized clinical trial. AB - Women with persistent HPV infections have increased risk of progressive CIN lesions. Transmission of HPV between sexual partners might maintain viral infection and, consequently, may influence the clinical course of CIN. We investigated the effect of condom use on regression of CIN lesions and on clearance of HPV. Women with CIN and their male sexual partners were randomized for condom use (condom group n = 72 and noncondom group n = 76). They were conservatively managed and followed every 3-6 months by colposcopy, cytology and HPV testing by GP5+/6+ PCR. Baseline cervical biopsy specimens were taken. Median follow-up time for women was 15.2 months (range 3.0-85.4). Outcomes of interest were clinical regression of CIN at colposcopy and clearance of HPV. Outcomes were assessed in 64 women of the condom group and 61 women of the noncondom group. Women in the condom group showed a 2-year cumulative regression rate of 53% vs. 35% in the noncondom group (p = 0.03). The 2-year cumulative rates of HPV clearance were 23% vs. 4%, respectively (p = 0.02). Although lower regression rates were found if women were HPV-positive and had > or =CIN2 lesions at baseline, effects of condom use were found both in women with CIN1 and in women with > or =CIN2 lesions. Condom use promotes regression of CIN lesions and clearance of HPV. PMID- 14566833 TI - Comparison of age at first full-term pregnancy between women with breast cancer and women with benign breast diseases. AB - Benign breast diseases have a broadly similar risk profile to that of breast cancer, possibly reflecting a similar underlying endocrine milieu. We have hypothesized that a crucial distinction between breast cancer and benign breast diseases is that mammary gland terminal differentiation has not been successfully accomplished among women who tend to develop breast cancer. From October 2001 to December 2002, information concerning breast cancer risk factors and sociodemographic characteristics was collected from 174 women with breast cancer and 116 women with benign breast diseases, all 30 years old or older, who were histologically diagnosed at a major prevention center in Athens, Greece. Among the examined breast cancer risk factors, only age at first full-term pregnancy was significantly associated with the odds of having breast cancer rather than benign breast disease, and the association was evident among premenopausal [odds ratio (OR) per 5 years = 1.76, 95% confidence interval (CI) 1.10-2.93] and postmenopausal (OR = 2.10, 95% CI 1.16-3.71) women, as well as among all women (OR = 1.93, 95% CI 1.34-2.70). There was no evidence that any of the remaining breast cancer risk factors could discriminate between breast cancer and benign breast diseases. We conclude that early age at first pregnancy may convey substantial protection against breast cancer risk among women with benign breast diseases, probably operating through induction of terminal differentiation of mammary gland cells. The finding is accentuated by the fact that women with benign breast diseases are already at a relatively high risk for breast cancer. PMID- 14566834 TI - Generation of a highly stable, internalizing anti-CD22 single-chain Fv fragment for targeting non-Hodgkin's lymphoma. AB - The generation of a single chain Fv (scFv) fragment derived from the anti-CD22 monoclonal antibody LL2 resulted in a molecule with good antigen binding but very poor stability properties, thus hampering its clinical applicability. Here we report on the construction of an engineered LL2 scFv fragment by rational mutagenesis. The contribution of uncommon wild-type sequence residues for providing stability to the conserved common core structure of immunoglobulins was examined. Aided by computer homology modeling, 3 destabilizing residues within the core of the wild-type VH domain were identified. Owing to the conserved nature of the buried core structure, mutagenesis of these sites to respective consensus residues markedly stabilized the molecule but did not influence its antigen binding properties: the engineered scFv MJ-7 exhibited exceptional biophysical stability with a half-life not reached after 6 days of incubation in human serum at 37 degrees C, while fully retaining the epitope specificity of the monoclonal antibody, and antigen binding affinity of the wild-type scFv. Furthermore, both the monoclonal antibody LL2 and the engineered scFv fragment became fully internalized after only 30 min of incubation at 37 degrees C with CD22+ tumor cells. These properties predict scFv MJ-7 could become a novel powerful tool to selectively deliver cytotoxic agents to malignant CD22+ cells. PMID- 14566835 TI - Piroxicam selectively inhibits the growth of premalignant and malignant human oral cell lines by limiting their progression through the S phase and reducing the levels of cyclins and AP-1. AB - Studies have shown that nonsteroidal antiinflammatory drugs (NSAIDs) reduce the risk of and mortality from a variety of cancers. Although cyclooxygenase (COX) dependent and -independent pathways may be involved, the mechanisms responsible for these effects remain unknown. In our study, we found that piroxicam inhibited cell growth in premalignant and malignant, but not normal, human oral epithelial cell lines in a concentration- and time-dependent manner. After 6 days of exposure, the concentration that inhibited growth by 50% was 181 and 211 microM for premalignant and malignant cells, respectively. Piroxicam did not induce apoptosis. The growth inhibitory effect was COX and PGE2 independent. Adding PGE2 or infecting cells with a COX-1 transgene did not abrogate piroxicam-induced growth inhibition. After treatment of the premalignant and malignant cell lines with piroxicam, cells accumulated in the S phase of the cell cycle. Upon removal of piroxicam, cells entered the G2 phase. The S phase block was accompanied by a reduction in the protein levels of cyclin A, cyclin B1, cyclin D1, cdc2, PCNA and the c-jun AP-1 component. Therefore, piroxicam may exert its growth inhibitory effects selectively on the premalignant and malignant human oral epithelial cells lines via signaling pathways regulating the progression of cells through the S phase of the cell cycle. PMID- 14566836 TI - 15-hydroxy-eicosatetraenoic acid arrests growth of colorectal cancer cells via a peroxisome proliferator-activated receptor gamma-dependent pathway. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) inhibits cell growth via promoting apoptosis. Human colorectal cancer tissues had abundant PPARgamma but the incidence of apoptosis was very low, suggesting a defect in the PPARgamma pathway. Here, we found that 15-hydroxy-eicosatetraenoic acid (15S-HETE), an endogenous ligand for PPARgamma, was significantly decreased in the serum of patients with colorectal cancer. Treatment of colon cancer cells with 15S-HETE inhibited cell proliferation and induced apoptosis, which was preceded by an increase in TGF-beta-inducible early gene (TIEG) and a decrease in Bcl-2. The action of 15S-HETE could be blocked when PPARgamma was suppressed. Overexpression of Bcl-2 prevented the apoptosis. The levels of TIEG and 15-lipoxygenase (15 LOX), the enzyme responsible for 15S-HETE production, was decreased in colorectal cancer. Therefore, colorectal cancer is associated with decreased 15S-HETE. Treatment of colon cancer cells with 15S-HETE inhibits cell proliferation and induces apoptosis in a PPARgamma-dependent pathway involving augmentation of TIEG and reduction of Bcl-2 expression. PMID- 14566837 TI - Nonsteroidal anti-inflammatory drugs inhibit growth of human neuroendocrine tumor cells via G1 cell-cycle arrest. AB - Therapeutic options to inhibit growth of human NETs of the GEP system are limited. Since NSAIDs might provide an antiproliferative treatment alternative with acceptable toxicity, we examined the effects of different NSAIDs on growth and survival in a representative set of human GEP NET cell lines. Growth and apoptosis were determined based on cell numbers, cell-cycle analyses, kinase assays, DNA fragmentation and PARP cleavage. Expression of COX and cell cycle regulatory molecules was examined by immunoblotting and reporter gene assays. Depending on the drug and cell line investigated, NSAID treatment resulted in profound growth inhibition of GEP NET cells. Growth-inhibitory effects were achieved with either COX-2 selective (NS398) or unselective (indomethacin, sulindac) compounds. Cell-cycle analyses documented a G1 arrest in NSAID-treated GEP NET populations. In addition, 100 microM sulindac or indomethacin induced apoptosis. All 3 COX inhibitors prevented CDK-2 activation. In parallel to the NSAID-mediated reduction of CDK-2 activity, p21(cip-1) promoter activity and cellular p21(cip-1) levels increased and p21(cip-1) was sequestered into CDK-2 complexes. Thus, the G1 arrest likely resulted from p21(cip-1)-dependent inhibition of CDK-2 activity. At therapeutically relevant concentrations, sulindac significantly reduced GEP NET cell numbers, whereas IFN-alpha and octreotide remained ineffective. The extent of growth inhibition in GEP NETs was comparable to the antiproliferative effects of sulindac in established NSAID sensitive cell models. NSAIDs acted as potent antiproliferative agents in GEP NET cells via G1 cell-cycle arrest and might therefore offer a therapeutic alternative to current treatment modalities. PMID- 14566838 TI - Loss of FHIT protein expression is a marker of adverse evolution in good prognosis localized breast cancer. AB - The FHIT tumor suppressor gene, which encompasses the fragile site FRA3B at 3p14.2, is altered frequently in many types of human cancers. To determine its importance as a prognostic marker in breast cancer, the expression of the FHIT protein was studied in a series of 452 breast carcinomas by using immunohistochemistry on sections of tissue microarrays. Three distinct levels of FHIT expression were observed: in 154 cases (34.1%) expression was unchanged as compared to normal level; in 78 (17.2%) no expression was found; in the remaining 220 cases (48.7%), expression was intermediate. Overall, two-thirds of the cases had abnormal levels of the protein. Absence of FHIT was significantly associated with a higher grade (p < 0.01) and absence of hormone receptors (p < 0.001). The patients were separated into Group I (153 node-negative good prognosis patients who did not receive adjuvant chemotherapy) and Group II (226 high-risk patients treated by adjuvant chemotherapy) according to the St.-Gallen conference consensus. The median follow-up was 48 months. Among Group I but not Group II patients, a multivariate analysis showed that FHIT expression was significantly associated with disease-free survival. The relative risk of recurrence for FHIT negative Group I patients was 2.37 (1.21-4.64; p = 0.03). Thus, among the patients who present with tumors of apparent good prognosis, FHIT is an independent prognostic factor that distinguishes a subgroup of patients who could benefit from adjuvant treatment. PMID- 14566839 TI - Identification of a sperm protein 17 CTL epitope restricted by HLA-A1. AB - Sperm protein 17 (Sp17) is a novel cancer-testis antigen. We previously reported the successful generation of Sp17-specific HLA-A1-restricted cytotoxic T lymphocytes (CTLs) from the peripheral blood of a healthy donor using a recombinant Sp17 protein. These CTLs were able to kill not only target cells pulsed with the recombinant protein but also fresh Sp17+ tumor cells. In the present study, we have identified a nonapeptide sequence within the Sp17 protein that is predicted to have a high binding affinity for the HLA-A1 molecules. We generated the synthetic nonapeptide and successfully propagated a peptide specific CTL line. We confirmed that peptide Sp17(103-111) (ILDSSEEDK) contains an Sp17 CTL epitope restricted by HLA-A1 and identified amino acid residues 104, 107, 109 and 110 as crucial for the CTL lysis. Our findings therefore provide the tool for the characterization of CD8 T-cell function in vivo and generation of epitope-specific treatment strategies. PMID- 14566840 TI - IL-1B promoter polymorphism and Epstein-Barr virus in Dutch patients with gastric carcinoma. PMID- 14566841 TI - Second primary cancers after thymoma. PMID- 14566843 TI - Tumor suppressor genes and the two-hit model of recessive oncogenesis: celebrating Alfred Knudson's 80th birthday. PMID- 14566844 TI - Cancer genetics through a personal retrospectroscope. PMID- 14566845 TI - A tribute to Alfred G. Knudson. PMID- 14566846 TI - The two-hit model for Wilms' tumor: where are we 30 years later? PMID- 14566847 TI - A personal and professional memoir of Alfred Knudson. PMID- 14566848 TI - Multistage carcinogenesis and the incidence of human cancer. AB - We consider the implications of multistage carcinogenesis for the incidence of cancer in human populations. When clonal expansion of partially altered cells is properly accounted for, we find it unnecessary to invoke genomic instability as an early event in malignant transformation. Environmental agents that increase the rate of clonal expansion of partially altered cells are efficient carcinogens. As a corollary, intervention strategies that decrease this rate are to be preferred to strategies that decrease the rate of early mutational events in carcinogenesis. PMID- 14566849 TI - Search for unknown tumor-antagonizing genes. AB - Following the ingenious prediction of Alfred Knudson in 1971, the first tumor suppressor gene, RB1, has been isolated. Its product, the RB1 protein, was found to play a major role in the control of the cell cycle. The loss of heterozygosity (LOH) technique, introduced by Cavenee and colleagues, was an important milestone toward the confirmation of Knudson's hypothesis and the identification of the gene. Subsequently, the LOH technique has provided important clues that have led to the discovery of other tumor suppressor genes. Most of them play important roles in the regulation of the cell cycle and/or of apoptosis. Circumstantial evidence suggests that still other and perhaps many unknown genes may participate in the protection of the organism against malignant growth. The numerous genome losses in tumors, detected by LOH, comparative genomic hybridization, and by cytogenetic techniques, support this possibility. The early work of one of us (G.K.), together with Henry Harris and Francis Wiener, had shown that the malignant phenotype can be suppressed by hybridizing malignant with low- or non tumorigenic cells. However, analysis of this phenomenon failed to assign the inhibition of tumorigenicity to any particular gene. We have pursued the search for new tumor-antagonizing genes with two unconventional approaches, focusing on human chromosomal subband 3p21.3, a region frequently targeted by cytogenetically detectable deletions. We have detected four clusters of candidate tumor suppressor genes at 3p21.3 by a combination of deletion mapping and the "elimination test." These findings raise the question whether the number and variety of genes that may contribute to the defense against uncontrolled proliferation may have been underestimated. PMID- 14566850 TI - The recessive nature of dominance. PMID- 14566851 TI - Alfred Knudson: the importance of a visionary who enables scientists. PMID- 14566852 TI - Knudson's hypothesis and the TP53 revolution. PMID- 14566853 TI - Targeting tumor cells by enhancing radiation sensitivity. AB - The work of Al Knudson created the paradigm in which we see cancer as a result of the accumulation of multiple mutations. Our goal has been to exploit these mutations to develop strategies to enhance therapy for cancer by targeting the malignant cell while sparing the normal tissue. In studying the RAS oncogene, we observed that its expression when activated resulted in enhanced radioresistance. Conversely, inhibition of RAS made cells with activated RAS more radiosensitive. Hence, we postulated that it would be possible to sensitize tumors with RAS mutations to radiation without affecting the sensitivity of the normal tissue in patients with such tumors. This proved to be the case in animal models and has led to current clinical trials. These studies raised the question of identifying the downstream effectors of RAS that are responsible for altering the radiosensitivity of cells. We have found that phosphoinositide-3-kinase (PI3 kinase) is a critical component of this pathway. Blocking PI3 kinase enhanced the radiation response in vitro or in vivo of cells actively signaling through that pathway, but did not affect cells not actively signaling through PI3 kinase at the time of irradiation. Identification of tumors with active signaling in this pathway by immunohistochemical staining for phosphorylated AKT, the downstream target of PI3 kinase correlated with those patients for which radiation failed to achieve local control. Thus, characterization of the active signaling pathways in a given tumor might enable the selection of patients likely to respond to radiation. Pathways upstream from RAS may also be useful targets to consider for enhancing radiation therapy. Epidermal growth factor receptor (EGFR), which is upstream of PI3 kinase, may also mediate resistance through a common pathway. In addition to EGFR and RAS, PTEN can also regulate the PI3 kinase pathway. Identifying a common signal for EGFR, RAS, and PTEN that results in radiation resistance may uncover targets for developing molecular-based radiosensitization protocols for tumors resistant to radiation and thus lead to improvement of local control. PMID- 14566854 TI - Poly(ADP-ribose) and carcinogenesis. AB - Poly(ADP-ribose) and poly(ADP-ribose) polymerase (PARP) were discovered about 40 years ago, but their significance was not well elucidated until recently. In the early stage of the history of PARP, the presence of antibodies in the sera of human patients with lupus erythematosus indicated its natural occurrence. PARP, as well as the degrading enzyme, poly(ADP-ribose) glycohydrolase (PARG), are present in most eukaryotes except for yeasts. Studies that used inhibitors of PARP indicated the involvement of PARP and poly(ADP-ribose) in DNA damage repair, and eventually PARP was purified and the gene was cloned. Molecular analysis then revealed various functional domains, such as the one for binding to strand breaks of DNA. Parp-1-deficient and Parg-deficient cells showed, in general, enhanced sensitivity to the lethal effects of ionizing radiation and alkylating agents. Parp-1 knockout mouse embryonic stem cells developed into teratocarcinoma-like tumors when injected subcutaneously into nude mice, these tumors featuring giant cells similar to syncytiotrophoblastic giant cells with hyperploidy. Parp-1 was also found in centrosomes, suggesting that poly(ADP-ribose) and PARP-1 are functionally involved in the maintenance of chromatin structure and the equal distribution of chromosomes into daughter cells. Intriguing findings on the real biological significance continue to be generated, with new light shed on mechanisms of carcinogenesis and pointing to novel cancer treatments. Highlights during the last four decades of studies by laboratories focusing on poly(ADP ribose)/PARP, including our own, are condensed and summarized in this review. PMID- 14566855 TI - Uterine leiomyoma in the Eker rat: a unique model for important diseases of women. AB - Eker rats carry a defect in the Tsc-2 tumor suppressor gene and female Eker rats develop uterine leiomyoma with a high frequency. The presentation, response to hormones and molecular alterations in these mesenchymal smooth muscle tumors, closely resembles their cognate human disease. Female rats and tumor-derived cell lines from Eker rat leiomyomas (ELT lines) have been developed as an in vivo/in vitro model system for preclinical studies to identify novel therapeutic agents for this disease and for studying disease pathogenesis. In addition to serving as a model for uterine leiomyoma, Eker rats have proven valuable for studying lymphangioleiomyomatosis, a related proliferative smooth muscle disease of women. PMID- 14566856 TI - Hereditary renal carcinogenesis fitting Knudson's two-hit model: genotype, environment, and phenotype. AB - Cancer is a heritable disorder of somatic cells. Environment and heredity both operate in the origin of human cancer. The Eker (Tsc2 gene mutant) rat model of hereditary renal carcinoma (RC) is an example of Mendelian dominantly inherited predisposition to a specific cancer in an experimental animal. To the best of our knowledge, this was the first isolation of a Mendelian dominantly predisposing cancer gene in a naturally occurring animal model. Recently, we discovered a new hereditary renal carcinoma in the rat in Japan, and the rat was named the "Nihon" rat, and its predisposing (Nihon) gene could be a novel renal tumor suppressor gene. We present these unique models, comparing these two predisposing genes (both are located on rat chromosome 10), for the study of problems in carcinogenesis, for instance, species-specific difference in tumorigenesis, cell stage and tissue/cell-type specific tumorigenesis, multistep carcinogenesis, modifier gene(s) in renal carcinogenesis, cancer prevention, and the development of the therapeutic treatments that can be translated to human patients, as well as how environmental factors interact with cancer susceptibility gene(s). PMID- 14566857 TI - Multiple roles of the tuberous sclerosis complex genes. AB - Soon after proposing the "two-hit" hypothesis for tumorigenesis, Knudson pursued further experimental validation of the concept by using a rat model of dominantly inherited renal tumor. Today, the Eker rat is one of the best characterized models of tuberous sclerosis complex (TSC) and has been used extensively for study of the function of the TSC2 tumor suppressor gene. Along with TSC1, these two genes behave as expected for tumor suppressor genes with evidence for loss of heterozygosity in tumors and suppression of growth when expressed in proliferating cells. Despite much experimental work, the mechanisms of these genes have remained elusive until recently. This review summarizes some of the current concepts in our understanding of the biological and biochemical function of the TSC genes. PMID- 14566858 TI - Metastasis of benign tumor cells in tuberous sclerosis complex. AB - Lymphangiomyomatosis (LAM) is a life-threatening lung disease affecting almost exclusively young women. Histologically, LAM is characterized by the diffuse, bilateral proliferation of abnormal smooth muscle cells and cystic degeneration of the lung parenchyma. LAM can occur as an isolated disorder (sporadic LAM), or in women with tuberous sclerosis complex (TSC-LAM). Patients with both sporadic LAM and TSC-LAM often have benign renal angiomyolipomas. The smooth muscle cells within the angiomyolipomas are very similar to the smooth muscle cells in pulmonary LAM. Genetic data suggest that pulmonary LAM is the result of a highly unusual disease mechanism: the metastasis of benign cells. If LAM is the result of metastasis, it is remarkable that the metastasis occurs in women, but not in men. In this review, I discuss the genetic data supporting this metastatic model for LAM. The implications of the model for the functions of the TSC1 and TSC2 gene products, hamartin and tuberin, respectively, will also be considered. Hamartin and tuberin may play functional roles in the suppression of cell migration and/or metastasis, possibly through their regulation of the small GTPase Rho. PMID- 14566859 TI - Genetic hits and mutation rate in colorectal tumorigenesis: versatility of Knudson's theory and implications for cancer prevention. AB - The multistep model of carcinogenesis is now widely accepted for colorectal cancer and other common epithelial tumors of the adult. The challenges ahead are to define the number of genetic hits necessary for conversion of a normal cell into a cancer cell and to determine whether the observed increase in the mutation rate (mutator phenotype) is required. The beauty of Knudson's two-hit theory is its ability to explain diverse experimental situations and guide specific predictions, including some directly relevant to cancer prevention. PMID- 14566860 TI - NF2: the wizardry of merlin. AB - Neurofibromatosis type II (NF2) is an autosomal dominant cancer syndrome characterized by the formation of tumors of the nervous system, particularly schwannomas and meningiomas. The NF2 gene is also implicated in the development of sporadic schwannomas and meningiomas, as well as tumor types seemingly unrelated to the NF2 disorder, such as malignant mesotheliomas. Inactivation of NF2 occurs by a "two-hit" mechanism, as proposed by Al Knudson, and the NF2 gene behaves as a classical tumor suppressor gene. The NF2 gene product, merlin, exhibits homology with the ezrin-radixin-moesin family of membrane-cytoskeleton linking proteins. During the past several years, there has been intensive investigation aimed at elucidating the mechanisms underlying merlin's functions. In this review, we summarize the involvement of NF2 inactivation in tumorigenesis. We also discuss observations implicating merlin in cell motility and cell proliferation, with a focus on recent findings linking merlin to Rac signaling. PMID- 14566863 TI - Benzotriazole: an ideal synthetic auxiliary. AB - Benzotriazole is a synthetic auxiliary that offers many advantages. It is inexpensive, odorless, and stable. A benzotriazole group is easily introduced, activates molecules towards numerous transformations, and can be removed easily at the end of the reaction sequence. This Concept provides some recent examples of the synthetic application of benzotriazole methodology and is intended to draw attention to the versatile applications of benzotriazole in organic chemistry. PMID- 14566864 TI - Folding versus self-assembling. AB - Model foldable polymers with sequences of rigid hydrophobic chromophores and flexible hydrophilic tetra(ethylene glycol) were synthesized and used as a paradigm for studying molecular-folding and self-assembly phenomena. Our results demonstrate that intramolecular association or folding prevails over intermolecular interaction or self-assembling in the concentration region from 1 microM to 0.1 M. Importantly, folded polymeric nanostructures have absorption and fluorescence properties that are distinct from those of unfolded polymers or free monomers. We hypothesize that the origins of folding and self-assembly come from interactions between molecular units, and that the key parameter that regulates the on-and-off of such interactions is the distance R separating the two molecular units. Each molecular unit produces a characteristic force field, and when another molecular unit enters this field, the probability that the two units will interact increases significantly. A preliminary estimate of the radius of such a force field for the perylene tetracarboxylic diimide chromophore is about 90-120 A. As a result, phenomena associated with folding or self-assembly of molecular species are observed when these conditions are met in solution. PMID- 14566865 TI - Rigid, conjugated, fluoresceinated thymidine triphosphates: syntheses and polymerase mediated incorporation into DNA analogues. AB - Syntheses of a unique set of energy transfer dye labeled nucleoside triphosphates, compounds 1-3, are described. Attempts to prepare these compounds were only successful if the triphosphorylation reaction was performed before coupling the dye to the nucleobase, and not the other way around. Compounds were prepared as both the 2'-deoxy (a) and 2',3'-dideoxy- (b) forms. They feature progressively longer rigid conjugated linkers connecting the nucleobase and the hydroxyxanthone moiety. UV spectra of the parent nucleosides 12-14 show that as the length of the linker increases so does the absorption of the donor in the 320 330 nm region, but with relatively little red-shift of the maxima. Fluorescence spectra of the same compounds show that radiation in the 320-330 nm region results in predominant emission from the fluorescein. When the linker is irradiated at 320 nm, the only significant emission observed corresponds to the hydroxyxanthone part of the molecules at 520 nm; this corresponds to an effective Stokes' shift of 200 nm. As the absorption at 320-330 nm by the linker increases with length, so does the intensity of the fluorescein emission. A gel assay was used to gauge relative incorporation efficiencies of compounds 1-3, dTTP, ddTTP, and 6-TAMRA-ddTTP. Throughout, the thermostable polymerase TaqFS was used, as it is the one most widely applied in high throughput DNA sequencing. This assay showed that only compounds 3 were incorporated efficiently; these have the longest linkers. Of these, the 2'-deoxy nucleoside 3 a was incorporated and did not prevent the polymerase from extending the chain further. The 2',3'-dideoxy nucleoside 3 b was incorporated only about 430 times less efficiently than ddTTP under the same conditions, and caused chain termination. The implications of these studies on modified sequencing protocols are discussed. PMID- 14566866 TI - In the twilight zone between [2]pseudorotaxanes and [2]rotaxanes. AB - A [2]pseudorotaxane, based on a semi-dumbbell-shaped component containing asymmetrically substituted monopyrrolotetrathiafulvalene and 1,5-dioxynaphthalene recognition sites for encirclement by cyclobis(paraquat-p-phenylene) and with a "speed bump" in the form of a thiomethyl group situated between the two recognition sites, has been self-assembled. This supramolecular entity is a mixture in solution of two slowly interconverting [2]pseudorotaxanes, one of which is on the verge of being a [2]rotaxane at room temperature, allowing it to be isolated by employing flash column chromatography. These two [2]pseudorotaxanes were both characterized in solution by UV/Vis and (1)H NMR spectroscopies (1D and 2D) and also by differential pulse voltammetry. The spectroscopic and electrochemical data reveal that one of the complexes behaves wholly as a [2]pseudorotaxane, while the other has some [2]rotaxane character to it. The kinetics of the shuttling of cyclobis(paraquat-p-phenylene) between the monopyrrolotetrathiafulvalene and the 1,5-dioxynaphthalene recognition sites have been investigated at different temperatures. The shuttling processes, which are accompanied by detectable color changes, can be monitored using UV/Vis and (1)H NMR spectroscopies; the spectroscopic data have been employed in the determination of the rate constants, free energies of activation, enthalpies of activation, and the entropies of activation for the translation of cyclobis(paraquat-p-phenylene) between the two recognition sites. PMID- 14566867 TI - Molecular energy and electron transfer assemblies made of self-organized lipid porphyrin bilayer vesicles. AB - Novel molecular energy and electron transfer assemblies in vesicular form, which are made of self-organized amphiphilic porphyrins bearing phospholipid-like substituents (lipid-porphyrins), have been photochemically characterized. Tetraphenylporphyrin (TPP) derivatives with four dialkylphosphocholine groups [free-base (1 a), Zn(2+) complex (1 b), and Fe(3+) complex (1 c)] are spontaneously associated in water to form spherical unilamellar vesicles with a diameter of 100-150 nm. Exciton calculations based on the bilayered sheet model of 1 b, which has a porphyrin packing similar to that seen in the triclinic unit cell of the Zn(2+)TPP crystals, reproduced the Soret band bathochromic shift appearing in the aqueous solution of 1 b well. The UV/Vis absorption spectrum of the 1 a/1 b hybrid vesicles (molar ratio: 1/1) showed no electronic interaction between the two porphyrin chromophores in the ground state, but efficient intermolecular singlet-singlet energy transfer took place from the excited 1 b donors to the 1 a acceptor within the vesicle. Near-field scanning optical microspectroscopy of the 1 a/1 b vesicles on a graphite surface also showed only free-base porphyrin fluorescence. The efficiency of the energy transfer was 0.81 and the rate constant was 3.1 x 10(9) s(-1). On the other hand, protoporphyrin IX bearing two alkylphosphocholine propionates (2) was incorporated into the 1 a or 1 c bilayer vesicles (ca. 100 nm phi, molar ratio: 1 a/2 or 1 c/2=10). The UV/Vis absorption spectrum showed that 2 was successfully anchored into the fluid alkylene region of the membrane without stacking. Photoirradiation (lambda(ex): 390 nm) of the 1 c/2 vesicles in the presence of triethanolamine led a vectorial electron transfer from the outer aqueous phase to the membrane center, which allowed reduction of the ferric ion of the Fe(3+)TPP platform. PMID- 14566869 TI - Computational assessment of the electronic structure of 1-azacyclohexa-2,3,5 triene (3 delta 2-1H-pyridine) and its benzo derivative (3 delta 2-1H-quinoline) as well as generation and interception of 1-methyl-3 delta 2-1H-quinoline. AB - Treatment of a solution of 3-bromo-1-methyl-1,2-dihydroquinoline (9) and [18]crown-6 in furan or styrene with KOtBu followed by hydrolysis afforded a mixture of 1-methyl-1,2-dihydroquinoline (10) and 1-methyl-2-quinolone (11). If the reaction was performed in [D(8)]THF and the mixture was immediately analysed by NMR spectroscopy, 2-tert-butoxy-1-methyl-1,2-dihydroquinoline (17) was shown to be the precursor of 10 and 11. The structure of 17 is evidence for the title cycloallene 7, which arises from 9 by beta elimination of hydrogen bromide and is trapped by KOtBu to give 17 so fast that cycloadditions of 7 with furan or styrene cannot compete. Since this reactivity is unusual compared to the large majority of the known six-membered cyclic allenes, we performed quantum-chemical calculations on 8, which is the parent compound of 7, and the corresponding isopyridine 6 to assess the electronic nature of these species. The ground state of 6 was no longer an allene (6 a) but the zwitterion 6 b. In the case of 8, the allene structure 8 a is more stable than the zwitterionic form 8 b by only approximately 1 kcal mol(-1). These results suggest a high reactivity of 6 and 8 towards nucleophiles and explains the behaviour of 7. In addition to the ground states, the low-lying excited states of 6 and 8 were considered, which are represented by the diradicals 6 c and 8 c and, as singlets, lie above 6 b and 8 a by 19.1-24.8 and 14.4-17.7 kcal mol(-1), respectively. PMID- 14566870 TI - Pyrrole-appended derivatives of O-confused oxaporphyrins and their complexes with nickel(II), palladium(II), and silver(III). AB - Condensation of 2,4-bis(phenylhydroxymethyl)furan with pyrrole and p toluylaldehyde formed, instead of the expected 5,20-diphenyl-10,15-di(p-tolyl)-2 oxa-21-carbaporphyrin, a pyrrole addition product [(H,pyr)OCPH]H(2); this product can formally be considered as an effect of hydrogenation of 3-(2'-pyrrolyl)-5,20 diphenyl-10,15-di(p-tolyl)-2-oxa-21-carbaporphyrin ([(pyr)OCPH]H). The new oxacarbaporphyrinoid presents the (1)H NMR spectroscopy features of an aromatic molecule, including the upfield shift of the inner H21 atom. Insertion of NiCl(2) or PdCl(2) into [(H,pyr)OCPH]H(2) gave two structurally related organometallic complexes, [(pyr)OCP]Ni(II)] and [(pyr)OCP]Pd(II)], in which the metal ions are bound by three pyrrolic nitrogens and the trigonally hybridized C21 atom of the inverted furan. The reaction of [(H,pyr)OCPH]H(2) with silver(I) acetate yields a stable Ag(III) complex [(C(2)H(5)O,pyr)OCP]Ag(III)] substituted at the C3 position by the ethoxy and pyrrole moieties. The macrocyclic frame of [(H,pyr)OCPH]H(2) is conserved. Addition of trifluoroacetic acid to [(C(2)H(5)O,pyr)OCP]Ag(III)] yielded a new aromatic complex [(pyr)OCP]Ag(III)](+). The structures of [(pyr)OCP]Ni(II)] and [(C(2)H(5)O,pyr)OCP]Ag(III)] have been determined by X-ray crystallography. In both molecules the macrocycles are only slightly distorted from planarity and the nickel(II) and silver(III) are located in the NNNC plane. The dihedral angle between the macrocyclic and appended-pyrrole planes of [(pyr)OCP]Ni(II)] reflects the biphenyl-like arrangement with the NH group pointing out toward the adjacent phenyl ring on the C5 position. Tetrahedral geometry around the C3 atom was detected for [(C(2)H(5)O,pyr)OCP]Ag(III)]. The Ni[bond]C and Ag[bond]C bond lengths are similar to other nickel(II) or silver(III) carbaporphyrinoids where the trigonal carbon atom coordinates the metal ion. The trend detected in the (13)C chemical shifts for the appended-pyrrole resonances has been rationalized by the extent of effective conjugation between the macrocycle and the appended pyrrole moiety controlled by the hybridization of the C3 atom and the metal ion oxidation state. The dianionic or trianionic macrocyclic core of the pyrrole appended derivatives is favored to match the oxidation state of nickel(II), palladium(II), or silver(III), respectively. PMID- 14566868 TI - pH-sensitive modulation of the second hydration sphere in lanthanide(III) tetraamide-DOTA complexes: a novel approach to smart MR contrast media. AB - The lanthanide(III) complexes of three tetraamide DOTA bearing pyridyl, phenolic and hydroxypyridyl substituents have been studied by NMR, luminescence and cyclic voltammetry. The relaxivity profiles of the gadolinium complexes of the pyridyl and phenolic ligands were flat and essentially the same between pH 2 and 8. The hydroxypyridyl ligand, however, exhibited two regions of enhanced relaxivity. The small relaxivity enhancement (25 %) at lower pH (pH 2-4) has been attributed to an increase in the prototropic exchange of the coordinated water molecule while the slightly larger enhancement (84 %) at higher pH (pH 6-9) reflects deprotonation of the ligand amide protons. Deprotonation of the amides results in the formation of an intramolecular acid-base pair interaction with the phenolic protons and this, in turn, causes a highly organized second hydration sphere to come into effect, thereby increasing the relaxivity. The water relaxivity of the Gd(3+)-hydroxypyridyl complex is further enhanced upon binding to serum albumin. PMID- 14566871 TI - Synthesis, structure, and spectroscopy of phenylacetylenylene rods incorporating meso-substituted dipyrrin ligands. AB - The synthesis, structure, and spectroscopic characterization of a series of phenylacetylenylene rodlike molecules containing dipyrromethene (dipyrrin) ligands are described. The combination of the phenylacetylenylene groups with the porphyrinogenic dipyrrin moieties results in a rich absorption spectroscopy for these compounds, although the fluorescence of the phenylacetylenylene moiety is quenched by presence of the dipyrrin chelator. The Cu(2+) and Fe(3+) complexes of these ligands have been prepared and three of these compounds have been structurally characterized by using single-crystal X-ray diffraction. Unlike other octahedral metal-dipyrrin complexes described to date, one of the iron complexes demonstrates ideal threefold symmetry in the solid-state. The elongated structure and high symmetry of these complexes suggests the use of these meso substituted phenylacetylenylene ligands as an interesting class of extended, branched molecules for the construction of supramolecular architectures. PMID- 14566872 TI - Theoretical investigation of the photochemical C2-C6 cyclisation of enyne heteroallenes. AB - Herein we discuss computations that explain experimental results regarding a highly efficient triplet analogue of the C(2)-C(6) cyclisation of enyne heteroallenes recently discovered by Schmittel and co-workers.1 To shed some light on the reasons for the differences found between enyne-carbodiimides, enyne ketenimines and enyne-allenes, we have computed the reaction profiles of the C(2) C(6) and of the C(2)-C(7) cyclisations for various model compounds, assuming that the reactions take place on the lowest-lying triplet surfaces. Our results nicely explain the differences and the unexpected high efficiency found for the enyne carbodiimides. The differences between enyne-carbodiimides and enyne-ketenimines prove to be due to differences in the shapes of the corresponding triplet surfaces. In contrast to the enyne-carbodiimides, for which our calculations predict that a direct cyclisation to the biradical intermediates should occur after the vertical excitation, the enyne-ketenimines relax into a local minimum on the triplet surface. As a consequence, further reaction channels are opened. Our computations indicate that enyne-allene compounds do not react because the necessary excitation energy lies outside the range of the employed triplet photosensitizer. Finally, the close agreement between our results and the experimental findings indicates that the underlying reasons for the differences in the photochemical behaviour of enyne-carbodiimides, enyne-ketenimines and enyne-allenes are related to differences in the electronic structures of the parent systems, while substituent effects are less important. PMID- 14566873 TI - Absolute asymmetric synthesis of "chiral-at-metal" Grignard reagents and transfer of the chirality to carbon. AB - Two new six-coordinate Grignard reagents, cis-[(p-CH(3)C(6)H(4))MgBr(dme)(2)] (1) and cis-[MgCH(3)(thf)(dme)(2)]I (2), have been synthesized and their crystal structures have been determined. Both reagents are cis-octahedral and therefore chiral. They crystallize as conglomerates and racemize rapidly in solution. By utilizing these properties, the absolute asymmetric synthesis of specifically the Delta or the Lambda enantiomer was achieved for both Grignard reagents. Enantiopure 1 and 2 were then reacted with butyraldehyde or benzaldehyde to give the corresponding alcohol in up to 22 % enantiomeric excess. At -60 degrees C, the Grignard reagents crystallize as racemic phases instead of conglomerates. Consequently, the crystal structures of rac-cis-[(p CH(3)C(6)H(4))MgBr(dme)(2)].DME (3) and rac-cis-[MgCH(3)(thf)(dme)(2)]I (4) could be determined. PMID- 14566874 TI - Orthoesters versus 2-O-acyl glycosides as glycosyl donors: theorectical and experimental studies. AB - n-Pentenyl orthoesters (NPOEs) undergo routine acid catalyzed rearrangement into 2-O-acyl n-pentenyl glycosides (NPGs). The reactant and product can both function as glycosyl donors affording 1,2-trans linked glycosides predominantly. However, both donors differ in their rates of reactions, the yields they produce, and the nature of their byproducts, indicating that the NPOE/NPG pair may not be reacting through the same intermediates. We have therefore applied quantum chemical calculations using DFT methods and MP second order perturbation theory to learn more about orthoesters and their 2-O-acyl glycosidic counterparts. The calculations show that in the case of a manno NPG and NPOE pair, each donor goes initially to a different cationic intermediate. Thus, the former goes to a high energy oxocarbenium ion before descending to a trioxolenium ion in which the charge is distributed over the pyrano ring oxygen, as well as the carbonyl and ether oxygen atoms of the putative C2 ester. On the other hand, ionization of the NPOE produces a dioxolenium ion lying slightly above the more stable trioxolenium counterpart. For the gluco pair, the NPG also goes to a very high-energy oxocarbenium ion, which also descends to a trioxolenium ion. However, unlike the manno analogue, the gluco NPOE does not give a dioxolenium ion; indeed, the dioxolenium is not energetically distinguishable from the trioxolenium counterpart. The theoretical observations have been tested experimentally. Thus, it was found that with manno derivatives, the orthoester is a more reactive donor than the corresponding NPG donor, whereas, for gluco derivatives, there is no advantage to using one over the other, unless one resorts to carefully selected promoters. PMID- 14566875 TI - Radicaloid-type oxidative decomposition of beer bittering agents revealed. AB - trans-Isohumulones, dihydroisohumulones, tetrahydroisohumulones, and humulinones, which are important hop-derived flavor components of beer, were found, by using electrolysis of organic solutions, to be stable against oxidation, like weak acids; however, they are readily oxidized in their anionic forms as present in beer. Oxygen- and carbon-centered radicals were formed by oxidation and identified by using spin trapping under aerobic and anaerobic conditions, followed by EPR (electron paramagnetic resonance) spectroscopy. Generated radicals were reactive, most likely degrading into products lacking the tricarbonyl chromophore; this is typical of five-membered-ring hop derivatives. Thus, flavor-active beer constituents may degrade oxidatively in the absence of oxygen, thereby leading to reaction products that escape UV detection. PMID- 14566876 TI - Catalysis of oxo transfer to prochiral sulfides by oxovanadium(v) compounds that model the active center of haloperoxidases. AB - The catalytic properties of a new class of chiral vanadium compounds--[(S,S,S) VO(OMe)L1] (5), [(S,S)-VO(OMe)L2] (6), [(S,S)-VO(OMe)L3] (7), and [(R,R,R) VO(OMe)L4] (8), as well as the system VO(OiPr)(3)/(R,R,R)-H(2)L4 [H(2)L1=(S,S) bis(2-hydroxypropyl)-(S)-1-phenylethylamine, 1; H(2)L2=(S,S)-bis(2 hydroxypropyl)benzylamine, 2; H(2)L3=(S,S)-bis(2-hydroxypropyl)isopropylamine), 3; (H(2)L4)=(R,R)-bis(2-phenylethanol)-(R)-1-phenylethylamine, 4]--in the asymmetric oxidation of prochiral sulfides by organic hydroperoxides have been investigated. Particular attention has been paid to the factors that guide the discrimination between the two prochiral faces of the sulfides (methyl p-tolyl sulfide and benzyl phenyl sulfide), to steric implications stemming from the oxidant (cumyl hydroperoxide and tert-butyl hydroperoxide), and to the specific complex used. As an example, (S)-methyl p-tolyl sulfoxide was obtained in a 31 % enantiomeric excess by use of cumyl hydroperoxide as oxidant and complex 5 as the catalyst, after 150 min at 0 degrees C and with 100 % conversion of the sulfide. The crystal and molecular structures of 5 and 6 reveal the close relationship between these complexes and the active center of vanadate-dependent haloperoxidases: the vanadium is in a slightly distorted trigonal-bipyramidal environment with the nitrogen and the methoxy group in the axial positions, and the oxo and alkoxide functions of L2 and L3 are the plane. The presence and equilibrium situation of isomers of the catalysts in solution has been investigated by (51)V EXSY and variable-temperature multinuclear NMR spectroscopy. An intermediately formed peroxo (ROO(-)) vanadium complex was detected by (51)V NMR spectroscopy. PMID- 14566877 TI - Selective recognition of configurational substates of zinc cyclam by carboxylates: implications for the design and mechanism of action of anti-HIV agents. AB - The interaction of metal cyclams with carboxylate groups is thought to play an important role in their binding to the CXCR4 chemokine receptor and in their anti HIV activity. Here we report the synthesis of acetate, phthalate, perchlorate and chloride complexes of Zn(II) cyclam (1,4,8,11-tetraazacyclotetradecane). The X ray crystal structures of [Zn(cyclam)(phthalate)](n)(CH(3)OH)(2n) and [Zn(cyclam)(H(2)O)(2)](OAc)(2) contain octahedral Zn(II) centres. Phthalate acts as a bridging ligand in the former complex, binding through monodentate carboxylate groups, and giving rise to infinite chains in the lattice together with extensive hydrogen bonding between carboxylate donor oxygen atoms and amine and methanol acceptor atoms. The uncoordinated acetate groups and the aqua ligand in the acetate complex are also involved in a rich network of hydrogen bonds and this may account for the unusually long Zn[bond]O distance (2.27 A). In both crystalline complexes, the macrocycle adopts the trans-III (S,S,R,R) configuration. 1D (1)H NMR spectra of all four complexes have been fully assigned by a combination of 2D [(1)H, (1)H] COSY and TOCSY, and [(1)H, (13)C] and [(1 )H, (15)N] HSQC NMR data. In aqueous solution, the stable trans-III configuration found in the solid-state equilibrates slowly (hours at 298 K) with trans-I (R,S,R,S) and cis-V (R,R,R,R) configurations. The trans-III configuration is predominant in aqueous solution for both the chloride and perchlorate complexes, but for the acetate and phthalate complexes, the cis-V configuration dominates. Carboxylate groups appear to stabilize the cis-V configuration in solution through Zn(II) coordination and hydrogen bonding. Titration of the chloride Zn(II)-cyclam complex with acetate confirmed that carboxylates strongly induce formation of the cis-V configuration. This implies that carboxylates can exert a strong influence over configurational selectivity. Cyclam NH hydrogen bonding is prevalent both in the solid state and in solution, and is relevant to the anti HIV activity of Zn(II) and other metal cyclam complexes and to their ability to recognize the CXCR4 transmembrane co-receptor. PMID- 14566878 TI - Electrostatic calculation of the substituent effect: an efficient test on isolated molecules. AB - The energy of a disubstituted molecule has often been approximated by simple electrostatic formulas that represent the substituents as poles or dipoles. Herein, we test this approach on a new model system that is more direct and more efficient than testing on acid-base properties. The energies of 27 1,4 derivatives of bicyclo[2.2.2]octane were calculated within the framework of the density functional theory at the B3LYP/6-311+G(d,p) level; interaction of the two substituents was evaluated in terms of isodesmic homodesmotic reactions. This interaction energy, checked previously on some experimental gas-phase acidities, was considered to be accurate and served as reference to test the electrostatic approximation. This approximation works well in the qualitative sense as far as the sign and the order of magnitude are concerned: beginning with the strongest interaction between two poles, a weaker interaction between pole and dipole, and the weakest between two dipoles. However, all the electrostatic calculations yield energies that are too small, particularly for weak interaction, and this fundamental defect is not remedied by some possible improvements. In particular, variation of the effective permittivity would require a physically impossible value less than unity. The explanation must lie in a more complex distribution of electron density than anticipated in the electrostatic model. It also follows that possible conclusions about the transmission of substituent effects "through space" have little validity. PMID- 14566879 TI - Discrete and infinite cage-like frameworks with inclusion of anionic and neutral species and with interpenetration phenomena. AB - Complex [Ag(tpba)N(3)] (1) was obtained by reaction of novel tripodal ligand N,N',N"-tris(pyrid-3-ylmethyl)-1,3,5-benzenetricarboxamide (TPBA) with [Ag(NH(3))(2)]N(3). While the reactions between 1,3,5-tris(imidazol-1-ylmethyl) 2,4,6-trimethylbenzene (TITMB) and silver(I) salts with different anions and solvent systems give six complexes: [Ag(3)(titmb)(2)](N(3))(3).CH(3)OH.4 H(2)O (2), [Ag(3)(titmb)(2)](CF(3)SO(3))(2)(OH).5 H(2)O (3), [Ag(3)(titmb)(2)][Ag(NO(3))(3)]NO(3).H(2)O (4), [Ag(3)(titmb)(2)(py)](NO(3))(3).H(2)O (py=pyridine) (5), [Ag(3)(titmb)(2)(py)](ClO(4))(3) (6), and [Ag(3)(titmb)(2)](ClO(4))(3).CHCl(3) (7). The structures of these complexes were determined by X-ray crystallography. The results of structural analysis of complexes 1 and 2, with the same azide anion but different ligands, revealed that 1 is a twofold interpenetrated 3D framework with interlocked cage-like moieties, while 2 is a M(3)L(2) type cage like complex with a methanol molecule inside the cage. Entirely different structure and topology between 1 and 2 indicates that the nature of organic ligands affected the structures of assemblies greatly. While in the cases of complexes 2-7 with flexible tripodal ligand TITMB, they are all discrete M(3)L(2) type cages. The results indicate that the framework of these complexes is predominated by the nature of the organic ligand and geometric need of the metal ions, but not influenced greatly by the anions and solvents. It is interesting that there is a divalent anion [Ag(NO(3))(3)](2-) inside the cage 4 and an anion of ClO(4)(-) or NO(3)(-) spontaneously encapsulated within the cage of complexes 5, 6 and 7. PMID- 14566880 TI - Reaction of alkyn-1-yl(diorganyl)silanes with 1-boraadamantane: Si-H-B bridges confirmed by the molecular structure in the solid state and in solution. AB - 1-Boraadamantane 1 was treated with alkyn-1-ylsilanes 2 containing one or two Si[bond]H functions. Under mild conditions, the reaction gave 4-methylene-3 borahomoadamantane derivatives 4 quantitatively and selectively by 1,1 organoboration. An electron deficient Si-H-B bridge was present in the product. The analogous reaction of 1 with an alkyn-1-yl-disilane 3 gave the corresponding alkene derivative 5, however, without the Si-H-B bridge. Evidence for the Si-H-B bridge in 4 was given by IR data, an extensive set of NMR spectroscopical data ((1)H, (11)B, (13)C, (29)Si NMR) including various unusual isotope effects on chemical shifts and coupling constants, as well as from the molecular structure of one example, 4 e, in the solid state. The precursor of 4 e, alkyne 2 e, Ph(2)Si(H)C[triple bond]CSi(H)Ph(2), was also studied by X-ray analysis. PMID- 14566881 TI - GA and AG sequences of DNA react with cisplatin at comparable rates. AB - The sequence selectivity of the antitumor drug cisplatin (cis-[PtCl(2)(NH(3))(2)] (1)) between the 5'-AG-3' and 5'-GA-3' sites of DNA has been a matter of discussion for more than twenty years. In this work, we compared the reactivity of GA and AG sequences of DNA towards the aquated forms of cisplatin (cis [PtCl(NH(3))(2)(H(2)O)](+) (2), cis-[Pt(NH(3))(2)(H(2)O)(2)](2+) (3), and cis [Pt(OH)(NH(3))(2)(H(2)O)](+) (4)) using two sets of experiments. In the first, we investigated a DNA hairpin, whose duplex stem contained a TGAT sequence as the single reactive site, and determined the individual rate constants of platination with 2 and 3 for G and A in acidic solution. The rate constants at 20 degrees C in 0.1M NaClO(4) at pH 4.5+/-0.1 were 0.09(4) M(-1)s(-1) (G) and 0.11(3) M(-1)s( 1) (A) for 2, and 9.6(1) M(-1)s(-1) (G) and 1.7(1) M(-1)s(-1) (A) for 3. These values are similar to those obtained previously for an analogous hairpin that contained a TAGT sequence. The monoadducts formed with 2 by both GA purines are extremely long-lived, partly as a result of the slow hydrolysis of the chloro monoadduct at A, and partly because of the very low chelation rate (1.4 x 10( 5)s(-1) at 20 degrees C) of the aqua monoadduct on the guanine. In the second set of experiments, we incubated pure or enriched samples of 1, 2, 3, or 4 for 18-64 h at 25 degrees C with a 19 base pair (bp) DNA duplex, whose radiolabeled top strand contained one GA and one AG sequence as the only reactive sites. Quantification of the number of GA and AG cross-links afforded a ratio of about two in favor of AG, irrespective of the nature of the leaving ligands. These results disagree with a previous NMR spectroscopy study, and indicate that GA sequences of DNA are substantially more susceptible to attack by cisplatin than previously thought. PMID- 14566882 TI - Electronically tuned chiral ruthenium porphyrins: extremely stable and selective catalysts for asymmetric epoxidation and cyclopropanation. AB - We report the use of three enantiomerically pure and electronically tuned ruthenium carbonyl porphyrin catalysts for the asymmetric cyclopropanation and epoxidation of a variety of olefinic substrates. The D(4)-symmetric ligands carry a methoxy, a methyl or a trifluoromethyl group at the 10-position of each of the 9-[anti-(1,2,3,4,5,6,7,8-octahydro-1,4:5,8-dimethanoanthracene)]-substituents at the meso-positions of the porphyrin. Introduction of a CF(3)-substituent in this remote position resulted in greatly improved catalyst stability, and turnover numbers of up to 7500 were achieved for cyclopropanation, and up to 14,200 for epoxidation, with ee values typically >90 % and approximately equal to 80 %, respectively. In one example, the axial CO ligand at the ruthenium was exchanged for PF(3), resulting in the first chiral ruthenium porphyrin with a PF(3) ligand reported to date. In cyclopropanations with ethyl diazoacetate, the latter catalyst performed exceedingly well, and gave a 95 % ee in the case of 1,1 diphenylethylene as substrate. PMID- 14566883 TI - Unusual iron(III) ate complexes stabilized by Li-pi interactions. AB - Several iron(III) complexes incorporating diamidoether ligands are described. The reaction between [Li(2)[RN(SiMe(2))](2)O] and FeX(3) (X=Cl or Br; R=2,4,6-Me(3)Ph or 2,6-iPr(2)Ph) form unusual ate complexes, [FeX(2)Li[RN(SiMe(2))](2)O](2) (2, X=Cl, R=2,4,6-Me(3)Ph; 3, X=Br, R=2,4,6-Me(3)Ph; 4, X=Cl, R=2,6-iPr(2)Ph) which are stabilized by Li-pi interactions. These dimeric iron(III)-diamido complexes exhibit magnetic behaviour characteristic of uncoupled high spin (S= 5/2 ) iron(III) centres. They also undergo halide metathesis resulting in reduced iron(II) species. Thus, reaction of 2 with alkyllithium reagents leads to the formation of iron(II) dimer [Fe[Me(3)PhN(SiMe(2))](2)O](2) (6). Similarly, the previously reported iron(III)-diamido complex [FeCl[tBuN(SiMe(2))](2)O](2) (1) reacts with LiPPh(2) to yield the iron(II) dimer [Fe[tBuN(SiMe(2))](2)O](2) but reaction with LiNPh(2) gives the iron(II) product [Fe(2)(NPh(2))(2)[tBuN(SiMe(2))](2)O] (5). Some redox chemistry is also observed as side reactions in the syntheses of 2-4, yielding THF adducts of FeX(2): the one-dimensional chain [FeBr(2)(THF)(2)](n) (7) and the cluster [Fe(4)Cl(8)(THF)(6)]. The X-ray crystal structures of 3, 5 and 7 are described. PMID- 14566884 TI - Hydrogen adsorption in carbon nanostructures: comparison of nanotubes, fibers, and coals. AB - Single-walled carbon nanotubes (SWNT) were reported to have record high hydrogen storage capacities at room temperature, indicating an interaction between hydrogen and carbon matrix that is stronger than known before. Here we present a study of the interaction of hydrogen with activated charcoal, carbon nanofibers, and SWNT that disproves these earlier reports. The hydrogen storage capacity of these materials correlates with the surface area of the material, the activated charcoal having the largest. The SWNT appear to have a relatively low accessible surface area due to bundling of the tubes; the hydrogen does not enter the voids between the tubes in the bundles. Pressure-temperature curves were used to estimate the interaction potential, which was found to be 580+/-60 K. Hydrogen gas was adsorbed in amounts up to 2 wt % only at low temperatures. Molecular rotations observed with neutron scattering indicate that molecular hydrogen is present, and no significant difference was found between the hydrogen molecules adsorbed in the different investigated materials. Results from density functional calculations show molecular hydrogen bonding to an aromatic C[bond]C that is present in the materials investigated. The claims of high storage capacities of SWNT related to their characteristic morphology are unjustified. PMID- 14566885 TI - Photoresponsive supramolecular systems: self-assembly of azodibenzoic acid linear tapes and cyclic tetramers. AB - A new strategy to effect photoinduced control over molecular self-assembly is reported. This strategy uses the reversible trans-cis photoisomerization of a novel azobenzene system, where the trans- and cis-forms self-assemble into dramatically different higher-order structures. The trans-azobenzene form of this molecule associates into infinite hydrogen-bonded linear tapes, while the cis azobenzene form undergoes hydrogen-bonded self-assembly into cyclic tetramers. This results in a second level of association, where the cis-hydrogen-bonded supramolecular cycles ultimately form long, rod-like aggregates through stacking interactions. PMID- 14566886 TI - Bonding of multiple noble-gas atoms to CUO in solid neon: CUO(Ng)n (Ng=Ar, Kr, Xe; n=1, 2, 3, 4) complexes and the singlet-triplet crossover point. AB - Laser-ablated U atoms co-deposited with CO in excess neon produce the novel CUO molecule, which forms distinct Ng complexes (Ng=Ar, Kr, Xe) with the heavier noble gases. The CUO(Ng) complexes are identified through CO isotopic and Ng reagent substitution and comparison to results of DFT frequency calculations. The U[bond]C and U[bond]O stretching frequencies of CUO(Ng) complexes are slightly red-shifted from neon matrix (1)Sigma(+) CUO values, which indicates a (1)A' ground state for the CUO(Ng) complexes. The CUO(Ng)(2) complexes in excess neon are likewise singlet molecules. However, the CUO(Ng)(3) and CUO(Ng)(4) complexes exhibit very different stretching frequencies and isotopic behaviors that are similar to those of CUO(Ar)(n) in a pure argon matrix, which has a (3)A" ground state based on DFT vibrational frequency calculations. This work suggests a coordination sphere model in which CUO in solid neon is initially solvated by four or more Ne atoms. Up to four heavier Ng atoms successively displace the Ne atoms leading ultimately to CUO(Ng)(4) complexes. The major changes in the CUO stretching frequencies from CUO(Ng)(2) to CUO(Ng)(3) provides evidence for the crossover from a singlet ground state to a triplet ground state. PMID- 14566887 TI - Alpha-gamma hybrid peptides that contain the conformationally constrained gabapentin residue: characterization of mimetics of chain reversals. AB - The crystal structures of four dipeptides that contain the stereochemically constrained gamma-amino acid residue gabapentin (1-(aminomethyl)cyclohexaneacetic acid Gpn) are described. The molecular conformation of Piv-Pro-Gpn-OH (1), reveals a beta-turn mimetic conformation, stabilized by a ten atom C[bond]H...O hydrogen bond between the Piv CO group and the pro S hydrogen of the Gpn CH(2)[bond]CO group. The peptides Boc-Gly-Gpn-OH (2), Boc-Aib-Gpn-OH (3), and Boc Aib-Gpn-OMe (4) form compact, folded structures, in which a distinct reversal of polypeptide chain direction is observed. In all cases, the Gpn residue adopts a gauche,gauche (g,g) conformation about the C(gamma)[bond]C(beta) (theta(1)) and C(beta)[bond]C(alpha) (theta(2)) bonds. Two distinct Gpn conformational families are observed. In peptides 1 and 3, the average backbone torsion angle values for the Gpn residue are phi=98 degrees, theta(1)=-62 degrees, theta(2)=-73 degrees, and psi=79 degrees, while in peptide 2 and 4 the average values are phi=-103 degrees, theta(1)=-46 degrees, theta(2)=-49 degrees, and psi=-92 degrees. In the case of 1 and 3, an intramolecular nine-membered O[bond]H...O hydrogen bond is formed between the C[double bond]O of the preceding residue and the terminal carboxylic acid OH group. All four alpha-gamma dipeptide sequences yield compact folded backbone conformations; this suggests that the Gpn residue may be employed successfully in the design of novel folded structures. PMID- 14566888 TI - Synthesis and characterization of new biphenolate and binaphtholate rare-Earth metal amido complexes: catalysts for asymmetric olefin hydroamination/cyclization. AB - Monomeric diolate amido yttrium complexes [Y[diolate][N(SiHMe(2))(2)](thf)(2)] can be prepared in good yield by treating [Y[N(SiHMe(2))(2)](3)(thf)(2)] with either 3,3'-di-tert-butyl-5,5',6,6'-tetramethyl-1,1'-biphenyl-2,2'-diol (H(2)(Biphen)), 3,3'-bis(2,4,6-triisopropylphenyl)-2,2'-dihydroxy-1,1'-dinaphthyl (H(2)(Trip(2)BINO)) or 3,3'-bis(2,6-diisopropylphenyl)-2,2'-dihydroxy-1,1' dinaphthyl (H(2)(Dip(2)BINO)) in racemic and enantiopure form. The racemic complex [Y(biphen)[N(SiHMe(2))(2)](thf)(2)] dimerizes upon heating to give the heterochiral complex (R,S)-[Y(biphen)[N(SiHMe(2))(2)](thf)](2). The corresponding dimeric heterochiral lanthanum complex was the sole product in the reaction of H(2)(Biphen) with [La[N(SiHMe(2))(2)](3)(thf)(2)]. Single-crystal X-ray diffraction of both dimeric complexes revealed that the two Ln(biphen)[N(SiHMe(2))(2)](thf) fragments are connected through bridging phenolate groups of the biphenolate ligands. The two different phenolate groups undergo an intramolecular exchange process in solution leading to their equivalence on the NMR timescale. All complexes were active catalysts for the hydroamination/cyclization of aminoalkynes and aminoalkenes at elevated temperature, with [Y((R)-dip(2)bino)[N(SiHMe(2))(2)](thf)(2)] being the most active one giving enantioselectivities of up to 57 % ee. Kinetic resolution of 2 aminohex-5-ene proceeded with this catalyst with 6.4:1 trans selectivity to give 2,5-dimethylpyrrolidine with a k(rel) of 2.6. PMID- 14566889 TI - Evidence for the formation of singly bonded dimers during the reductive electrochemistry of methanofullerenes. AB - Four methanofullerene derivatives, with phosphonate or sulfone groups attached to a C(60) core through a Bingel procedure, were synthesized to probe their stability upon electrolytic reduction. Derivatives 1 and 2 are the most stable upon electroreduction and do not exhibit retro-cyclopropanation reactions until more than three electrons per C(60) derivative are transferred. The cyclopropane ring is then removed and C(60)(>CH(2))(n) (n=1-3) products result from reactions of the trianion of C(60) with the solvent, CH(2)Cl(2). The situation with diphosphonate 3 or phosphonatecarboxylate 4 is dramatically different. For 3, quantitative retro-cyclopropanation occurs when 2.8 e(-) per molecule are transferred. In the case of 4, when more than two electrons per molecule are transferred, there is evidence of the reversible formation of a very stable intermediate, which is oxidized at a potential 500 mV more positive than the first fullerene-based reduction of the parent compound. Electrolysis of a simple C(70)-Bingel monoadduct (5) also exhibits the formation of a similar intermediate. On the basis of cyclic voltammetry, ESR spectroscopy, and MALDI analysis of products, the intermediate observed during the electrolysis of compounds 4 and 5 is assigned to a dimeric structure. PMID- 14566890 TI - The first structural characterisation of a group 2 metal alkylperoxide complex: comments on the cleavage of dioxygen by magnesium alkyl complexes. AB - A new high-yield synthesis of [(PhCH(2))(2)Mg(thf)(2)] and [[(PhCH(2))CH(3)Mg(thf)](2)] via benzylpotassium has allowed a simple entry into benzylmagnesium coordination chemistry. The syntheses and X-ray crystal structures of both [(eta(2)-Me(2)NCH(2)CH(2)NMe(2))Mg(CH(2)Ph)(2)] and [eta(2) HC[C(CH(3))NAr'](2)Mg(CH(2)Ph)(thf)] (Ar'=2,6-diisopropylphenyl) are reported. The latter beta-diketiminate complex reacts with dioxygen to provide a 1:2 mixture of dimeric benzylperoxo and benzyloxo complexes. The benzylperoxo complex [[eta(2)-HC[C(CH(3))NAr'](2)Mg(mu-eta(2):eta(1)-OOCH(2)Ph)](2)] is the first example of a structurally characterised Group 2 metal-alkylperoxo complex and contains the benzylperoxo ligands in an unusual mu-eta(2):eta(1)-coordination mode, linking the two five-coordinate magnesium centres. The O[bond]O separation in the benzylperoxo ligands is 1.44(2) A. Reaction of the benzylperoxo/benzyloxo complex mixture with further [eta(2)-HC[C(CH(3))NAr'](2)Mg(CH(2)Ph)(thf)] results in complete conversion of the benzylperoxo species into the benzyloxo complex. This reaction, therefore, establishes the cleavage of dioxygen by this system as a two-step process that involves initial oxygen insertion into the Mg[bond]CH(2)Ph bond followed by O[bond]O/Mg[bond]C sigma-bond metathesis of the resulting benzylperoxo ligand with a second Mg[bond]CH(2)Ph bond. The formation of a 1:2 mixture of the benzylperoxo and benzyloxo species indicates that the rate of the insertion is faster than that of the metathesis, and this is shown to be consistent with a radical mechanism for the insertion process. PMID- 14566894 TI - Clinical anatomy of ligamentum nuchae. AB - In view of the disparities in the anatomical, clinical, and biomechanical literature this study was undertaken to reappraise the gross anatomy of ligamentum nuchae with the objective of providing an accurate anatomical basis for biomechanical studies and for clinical theories and practices. Ligamentum nuchae was studied by gross dissection in ten embalmed human adult cadavers and was found to be comprised of two structures: a dorsal raphe and a midline fascial septum. The dorsal raphe was formed by the interweaving of the right and left upper trapezius, splenius capitis, and rhomboid minor. It spanned the cervical spine and was firmly attached to the external occipital protuberance and to the spinous process of C7. The fascial septum consisted of dense connective tissue and ran ventrally from the midline raphe to be confluent with the interspinous ligaments and atlanto-axial and atlanto-occipital membranes. These findings agree with those anatomical descriptions that portray ligamentum nuchae as a two-part structure but not with those studies in which it is characterized as a strong ligament attaching to all cervical spinous processes. Importantly, the findings of this study emphasize the need for clinical and biomechanical literature to portray the gross anatomy of ligamentum nuchae accurately. PMID- 14566891 TI - Complementary nucleobase interaction enhances peptide-peptide recognition and self-replicating catalysis. AB - The availability of the complementary interaction of nucleobases for influencing the formation of peptide architectures was explored. Nucleobases were incorporated as additional recognition elements in coiled-coil peptides by employing nucleobase amino acids (NBAs), which are artificial L-alpha-amino gamma nucleobase-butyric acids. The effect of the base-pair interaction on intermolecular recognition between peptides was evaluated through a self replication reaction. The self-replication reactions of the peptides with complementary base pairs such as thymine-adenine or guanine-cytosine at the g-g' heptad positions were accelerated in comparison with those of the peptides with mismatched base pairs or without nucleobases. However, thymine-adenine pairs at the e-e' positions did not enhance the self-replication. In the presence of a denaturant, the enhancement effects of complementary base pairs on the reaction disappeared. Thermal denaturation studies showed that the thymine-adenine pairs contributed to stabilization of the coiled-coil structure and that the pairs at the g-g' positions were more effective than those at the e-e' positions. The peptide-peptide interaction was reinforced by complementary nucleobase interactions appropriately arranged in the peptide structure; these led to acceleration of the self-replication reactions. PMID- 14566895 TI - Comparison of mandibular landmarks from computed tomography and 3D digitizer data. AB - We recorded 3D coordinates for 28 mandibular landmarks from three-dimensional reconstructions of CT axial slices using the image analysis program eTDIPS. The images were acquired from a pediatric series of human mandibles (neonate to 13 years of age) from the Bosma collection (Shapiro and Richtsmeier, 1997, Am. J. Phys. Anthropol. 103:415-416). To test the accuracy of these coordinate data, we recorded the same 28 landmarks directly on the Bosma mandibles using a Polhemus 3Space digitizer. The directly digitized landmarks serve as a gold standard upon which to evaluate the eTDIPS data. Standard deviations of landmark placement using eTDIPS show a greater degree of variation compared to the data gathered using the digitizer, although this error is more heavily concentrated in certain types of landmarks. All possible linear distances between unique pairs of landmarks were calculated, and like linear distances were compared between the two data collection methods. The absolute difference for all like linear distances ranged from 0.001-3.9 mm (mean = 0.377 mm; SD = 1.136), with the eTDIPS data being consistently larger than the digitizer coordinates. This study demonstrates that landmark coordinate data can be reliably collected from digital CT images of the human mandible. We define a set of mandibular landmarks useful in evaluating the effects of craniofacial disorders, growth and other biological processes. PMID- 14566896 TI - Relationship of the common peroneal nerve and its branches to the head and neck of the fibula. AB - The common peroneal nerve (CPN) lies on the neck of the fibula, which forms the floor of the so-called 'fibular tunnel.' The tunnel entrance is a musculo aponeurotic arch derived from the soleus and peroneus longus muscles and it is here that the CPN is commonly compressed in cases of peroneal nerve palsy. This study aims to define the relationship of the CPN and its branches to the apex of the head of the fibula and to the tunnel, with special regard to possible sites of entrapment. The distances from the apex of the fibula to the opening of the fibular tunnel, the CPN bifurcation, and the exit point of the deep peroneal nerve (DPN) from the tunnel, were measured in 30 legs to ascertain possible sites of entrapment. The angle that the CPN subtended with the long axis of the fibula was measured to gauge the range of positions of the CPN at the neck of the fibula. An unyielding musculo-aponeurotic fibular arch at the entrance to the fibular tunnel was confirmed in all specimens. The DPN exited through a crescentic opening in the anterior intermuscular septum in all cases and no DPN branches were found in the lateral compartment in any specimen. The mean (+/-SD) distance from the apex of the head of the fibula to the opening of the fibular tunnel was 3.2 +/- 1.0 cm, to the CPN bifurcation was 3.8 +/- 0.9 cm, and to the DPN exit point was 7.0 +/- 1.5 cm. The mean angle subtended anteriorly from the long axis of the fibula by the CPN was 18.9 +/- 9.0 degrees. We recommend further study of the mean distances and reference angle in relation to fibular landmarks, for use in possible minimally invasive surgical procedures to decompress the fibular tunnel. PMID- 14566897 TI - Prevalence of osteophytes associated with the acromion and acromioclavicular joint. AB - Osteophytes as a stigma for shoulder impingement syndrome and osteoarthritis of the acromioclavicular (AC) joint were studied on the acromions and AC joints in 346 skeletons (230 males; 116 females), ranging in age from 15-100 years. Osteophytes were found on 28.9% (200/692) of the acromions, mostly on the anteroinferior aspect (54%; 108/200). Of these 28.9%, 87% (174/200) were of the traction type and 13% (26/200) of the claw type. Regarding the clavicles, 11.6% (80/692) had osteophytes on the articular facets of their lateral ends; all were of the traction type. There were statistically more osteophytes on the right than on the left side of both acromion and clavicle (P < 0.05). The occurrence of osteophytes and increasing age were significantly correlated (r = 0.65, P < 0.001). No gender differences were noted in the frequency of osteophytes on acromions or clavicles. These data should prove beneficial to clinicians in planning a proper course of treatment for patients suffering from painful conditions of the shoulder. PMID- 14566898 TI - The spinoglenoid ligament: an anatomic study. AB - The spinoglenoid ligament and its anatomic variations are described in 27 shoulders from 15 cadavers. In each shoulder one or two distinct spinoglenoid ligaments originated from the base of the spine of the scapula; they inserted on the neck of the scapula or the shoulder joint capsule. In the 19 shoulders in which only one spinoglenoid ligament was present, it inserted into the neck of the scapula in 14 cases and into the shoulder joint capsule in five instances. In the eight shoulders in which there were two ligaments, one inserted into the neck of the scapula and the other into the shoulder joint capsule. We did not observe any hypertrophic spinoglenoid ligaments that may have compressed the suprascapular nerve. PMID- 14566899 TI - Axillary arch: potential cause of neurovascular compression syndrome. AB - During routine bilateral dissection of 64 upper limbs in 32 adult human cadavers, four cases of an axillary arch, a muscular anomaly, were found. In one case, the axillary arch presented on both sides and in two cases it presented unilaterally on the right side. The innervation and relationships of the axillary arches are reported and the surgical significance of the anomaly is discussed. PMID- 14566900 TI - Interphalangeal sesamoid bones of the great toe: an anatomic variant demanding careful scrutiny of radiographs. AB - We describe a patient who was found to have two sesamoid bones at the interphalangeal joint of the right great toe after radiographs for dislocation of the joint. Recognition of the sesamoids required careful scrutiny of the films. Interphalangeal sesamoids may cause painful callosities plantar to the joint or may become incarcerated in a dislocated joint. Our patient made a good recovery. The presence of sesamoid bones on the medial and lateral sides of the joint was unusual. PMID- 14566901 TI - Muscular ventricular septal defect in an 89-year-old woman that was undetected during life. PMID- 14566902 TI - Outcomes of the gross and developmental anatomy teaching assistant experience. AB - During the first-year Gross and Developmental Anatomy Course at Mayo Medical School, third-year medical students volunteer as teaching assistants (TAs). Their responsibilities include preparing for dissection, instructing students during dissection, writing examination questions, and giving a lecture. To evaluate the academic and professional impact of this experience on former TAs, a survey instrument was developed, and was sent to former TAs from the past 17 years. Seventy-two percent of the surveys were returned. Most respondents (84%) indicated that the TA experience was beneficial during their third-year surgical rotation. Over 60% identified benefits during Neurology, Obstetrics and Gynecology, and Internal Medicine rotations. The majority (62%) indicated they regularly use the knowledge gained from the TA experience. Communication was the most highly ranked area of skill development, with 86% of respondents indicating gains in lecture effectiveness and 97% in one-on-one teaching. Among respondents, 32% entered surgical or radiological fields, roughly paralleling the non-TA medical student population. All respondents indicated that they would repeat the experience and recommend it to other medical students. Although benefits for long term career development have not yet been established, the Gross and Developmental Anatomy TA experience had perceived short-term benefits with respect to clinical rotations and teaching skill development. PMID- 14566903 TI - Unusual origin of the flexor digiti minimi brevis muscle. AB - An unusual origin of the flexor digiti minimi brevis muscle of the right hand was discovered during cadaveric dissection. The muscle originated from the anterior aspect of the transverse fibers of the distal antebrachial fascia and inserted onto the base of the fifth proximal phalanx. When traction was applied to the proximal portion of the muscle, flexion was produced at the fifth metacarpophalangeal joint. Other intrinsic muscles of the hand and the muscles of the flexor forearm compartment had normal morphology. Although muscular anomalies in the hypothenar region have been described, the muscular variant described here was distinct in its origin and size from those previously reported. Knowledge of anomalous muscles in the hand has important clinical significance in routine surgery and in determining associated pathology. PMID- 14566904 TI - Absence of the left inferior thyroid artery: clinical implications. AB - Dissection of an adult male cadaver revealed an absence of the left inferior thyroid artery; its usual area of distribution to the thyroid gland was supplied by the right inferior thyroid artery. Absence of the left inferior thyroid artery occurs in 1-6% of cases. The inferior thyroid artery arises commonly from the thyrocervical trunk, passes posterior to the carotid sheath and supplies the inferior pole of the corresponding lobe of the thyroid gland; its branches can course anterior or posterior to or between branches of the recurrent laryngeal nerve. During thyroid surgery it is imperative to identify the relationship of the inferior thyroid artery to the recurrent laryngeal nerve or to establish its absence because injury to the nerve can be a major complication; awareness of significant variations of the surgical anatomy of the thyroid gland is vital for preserving the integrity of important structures. PMID- 14566905 TI - Thoracoscopy: a new era for surgical anatomy. AB - In this new era of minimal access surgery, advances in optics and illumination have established thoracoscopic sympathectomy as a pre-eminent procedure, including a safe and efficient technique for upper limb sympathectomy. The success of thoracoscopy will doubtless ensure that a greater number of these procedures will be carried out and will put some of the daunting technical challenges posed by traditional open surgical procedures to rest. The thoracoscopic era affords the surgical anatomist a new challenge: to move the teaching of living anatomy to a higher level. PMID- 14566908 TI - Perspectives on the biostatistical sciences: A symposium in memory of Samuel W. Greenhouse. June 11, 2001, Bethesda, Maryland, USA. Proceedings. PMID- 14566909 TI - A tribute to Samuel W. Greenhouse. PMID- 14566910 TI - Sam Greenhouse's years at the Census Bureau and the UNRRA. AB - Sam Greenhouse joined the Census Bureau as a clerk at an interesting time period for the agency. The first use of sampling in the decennial census occurred in 1940. There was a major expansion of the amount of data collected. The organization of the Census Bureau underwent radical changes, including the growth of the Population Division of which Greenhouse was a part, and the Office of Statistical Assistant was established. After the 1940 census Greenhouse was also part of a new intercensal program which became the forerunner of today's Current Population Survey, and was on hand for the transition of the physical location of the agency from Washington D.C. to Suitland, Maryland. After America's entry into World War II, Sam Greenhouse left the Census Bureau and worked with the United Nations Relief and Rehabilitation Administration (UNRRA), designed to manage the support and eventual repatriation of refugees who would come under Allied control at the war's end. Published in 2003 by John Wiley & Sons, Ltd. PMID- 14566912 TI - Sam Greenhouse: his contributions as a consultant to the Food and Drug Administration. AB - This paper recounts contributions made by Sam Greenhouse to the Food and Drug Administration during his tenure as an advisory committee member and as chair of the committee. The events and topics are taken from available recollections and minutes and selectively describe a range of topic areas and issues about which Sam Greenhouse played a substantial leadership role. Published in 2003 by John Wiley & Sons, Ltd. PMID- 14566911 TI - Sam Greenhouse's scholarly contributions to human aging studies. PMID- 14566913 TI - The contributions of Sam Greenhouse to research on psychiatric diagnosis and psychopharmacology. AB - Sam Greenhouse began his involvement in mental disorders research in 1954 when appointed chief of the Theoretical Statistics and Mathematics section at the National Institute of Mental Health. He remained with the NIMH until 1966. Despite moving on to several other positions at the NIH and at the university during the ensuing years, he continued as a consultant to NIMH investigators. He also participated actively as an advisor and co-investigator on a number of important collaborative research programmes launched by the Institute in the 1970s and 1980s. His contributions to the design and methodology of the first clinical trials of drugs for the treatment of schizophrenia, to research aimed at revising the national and international classification systems for the mental disorders, and his participation in the planning of the first attempt to use the collaborative research model to test hypotheses about the genesis of a specific mental disorder (depression), are described. Finally, the signal importance of the 'profile analysis of variance' method that he and Seymour Geisser developed, to research on personality and mental disorders, is examined in detail. The authors describe applications of the method in their own research on the classification of the mental disorders, predicting response to drug treatment and the variations in the expression of mental illness across different cultures. Sam worked in mental health during an era of revolutionary changes in the diagnosis and treatment of mental disorders. The field was acutely aware of his many contributions to the progress of research and his colleagues are very grateful to have had the opportunity to work with him. PMID- 14566914 TI - Some of Sam Greenhouse's contributions to statistical methods. AB - I briefly survey some areas of Sam Greenhouse's contributions to statistical methods before focusing on three examples. These examples illustrate Sam's ability to identify problems of practical importance and to make valuable contributions to their solutions. Many of his papers continue to provide important guidance and insight because he dealt with issues of enduring practical importance. PMID- 14566916 TI - On psychiatry, epidemiology, and statistics: a view from the 1950s and 1960s. PMID- 14566917 TI - The growth and future of biostatistics: (a view from the 1980s). PMID- 14566918 TI - The analysis of profile data--revisited. AB - We revisit a former problem that led to much commentary regarding a proposed use of testing whether profiles for several groups in a repeated measurement scenario were parallel. Here we review previous approaches and propose Bayesian model selection to indicate whether the profiles can be considered parallel and if so support an estimation procedure for the profile differences. PMID- 14566919 TI - Over-ruling a group sequential boundary--a stopping rule versus a guideline. AB - We evaluate the properties of group sequential procedures where the trial is continued even though the boundary for statistical significance (stopping) to demonstrate effectiveness has been crossed. In this case, one may buy-back the previously spent alpha probability to be re-spent or re-distributed at future looks. We show that such plans using an O'Brien-Fleming-like spending function have a negligible effect on the final type I error probability and on the ultimate power of the study. With a Pocock-like bound, however, there is a small additional loss in power. We also show that this approach can be simplified by using a fixed-sample size Z critical value for future looks after buying-back previously spent alpha, such as using a critical Z value of 1.96 for alpha=0.025. We show that this procedure preserves the type I error probability while incurring a minimal loss in power. In this sense, one still has a stopping boundary rather than simply a guideline. This concept is discussed relative to monitoring procedures for inferiority or futility, and cases where both an upper and lower boundary are employed. PMID- 14566920 TI - Group sequential large sample T2-like chi2 tests for multivariate observations. AB - In many studies, a K degree of freedom large sample chi2 test is used to assess the effect of treatment on a multivariate response, such as an omnibus T2-like test of a difference between two treatment groups in any of K repeated measures. Alternately, a K df chi2 test may be used to test the equality of K+1 groups in a single outcome measure. Jennison and Turnbull (Biometrika 1991; 78: 133-141) describe group sequential chi2 and F-tests for normal errors linear models, and Proschan, Follmann and Geller (Statist. Med. 1994; 13: 1441-1452) describe group sequential tests for K+1 group comparisons. These methods apply to sequences of statistics that can be characterized as having an independent increments variance covariance structure, thus simplifying the computation of the sequential variance covariance matrix and the resulting sequential test boundaries. However, many commonly used statistics do not share this structure, including a Liang-Zeger (Biometrika 1986; 73: 13-22) GEE longitudinal analysis with an independence working correlation structure and a Wei-Lachin (J. Amer. Statist. Assoc. 1984; 79: 653-661) multivariate Wilcoxon rank test, among others. For such analyses, this paper describes the computation of group sequential boundaries for the interim analysis of emerging results using K df tests that are expressed as quadratic forms in a statistics vector that is distributed as multivariate normal, at least asymptotically. We derive the elements of the covariance matrix of multiple successive K df chi2 statistics based on established theorems on the distribution of quadratic forms. This covariance matrix is estimated by augmenting the data from the successive interim analyses into a single analysis from which the component sequential tests and their variance-covariance matrix can then be extracted. Boundary values for the sequential statistics can then be computed using the method of Slud and Wei (J. Amer. Statist. Assoc. 1982; 77: 862 868) or using the alpha-spending function of Lan and DeMets (Biometrika 1983; 70: 659-663) with a surrogate measure of information. An example is presented using the analysis of repeated cholesterol measurements in a clinical trial. PMID- 14566921 TI - Quantifying epidemiologic risk factors using non-parametric regression: model selection remains the greatest challenge. AB - Logistic regression is widely used to estimate relative risks (odds ratios) from case-control studies, but when the study exposure is continuous, standard parametric models may not accurately characterize the exposure-response curve. Semi-parametric generalized linear models provide a useful extension. In these models, the exposure of interest is modelled flexibly using a regression spline or a smoothing spline, while other variables are modelled using conventional methods. When coupled with a model-selection procedure based on minimizing a cross-validation score, this approach provides a non-parametric, objective, and reproducible method to characterize the exposure-response curve by one or several models with a favourable bias-variance trade-off. We applied this approach to case-control data to estimate the dose-response relationship between alcohol consumption and risk of oral cancer among African Americans. We did not find a uniquely 'best' model, but results using linear, cubic, and smoothing splines were consistent: there does not appear to be a risk-free threshold for alcohol consumption vis-a-vis the development of oral cancer. This finding was not apparent using a standard step-function model. In our analysis, the cross validation curve had a global minimum and also a local minimum. In general, the phenomenon of multiple local minima makes it more difficult to interpret the results, and may present a computational roadblock to non-parametric generalized additive models of multiple continuous exposures. Nonetheless, the semi parametric approach appears to be a practical advance. PMID- 14566922 TI - The use of the 'reverse Cornfield inequality' to assess the sensitivity of a non significant association to an omitted variable. AB - Unlike randomized experimental studies, investigators do not have control over the treatment assignment in observational studies. Hence, the treated and control (non-treated) groups may have widely different distributions of unobserved covariates. Thus, if observational data are analysed as if they had arisen from a controlled study, the analyses are subject to potential bias. Sensitivity analysis is a technique for assessing whether the inference drawn from a study could be altered by a moderate 'imbalance', between the distribution of the covariates in different groups. In this paper, we examine the sensitivity analysis of the test of proportions in 2 x 2 tables from a new perspective: 'could a non-significant result have occurred because the treated group has a higher prevalence of an unobserved risk factor?'. The study was motivated by an analysis of the studies concerning with the possible effect of spermicide use on birth defects that were cited in a legal decision. PMID- 14566923 TI - Estimating population size and duplication rates when records cannot be linked. AB - The capture-recapture approach to estimating the size of a population is a well studied area of statistics. The number of distinct individuals, N(A) and N(B), on each of two lists, A and B, and the number common to both lists, N(AB), are used to form an estimate of the binomial probability of being on one of the lists, which then allows an estimate to be made of the size of the population. Critical to the method is an accurate count of N(AB). We consider situations in which this count is not available. Such problems arise in a variety of behavioural health contexts in which the need for protection of privacy may prevent sharing identifying information, so it is not possible to specifically match an individual who appears on one list with an individual on the other. Suppose that the birth dates and/or other demographics of individuals on each list are known. We introduce two methods for estimating the duplication rates and the size of the population. Conditioning on the set beta of birth dates of those on list B, N(A) and N(B), the maximum likelihood estimators (MLEs) and their variance are derived. The MLEs are based on the proportion of individuals on list A whose birth dates fall in beta. This approach is particularly useful if list B itself contains duplicates. The second model utilizes the full sample distribution of the birth dates. We generalize this approach to accommodate multiple demographic characteristics. The approaches are applied to the problem of estimating duplication rates and the population size of veterans who have mental illness in Kings County, NY. The data are lists of those receiving service from the Veterans Administration system and from providers funded or certified by the New York State Office of Mental Health. PMID- 14566924 TI - Clinical trials, epidemiology, and public confidence. AB - Critics in the media have become wary of exaggerated research claims from clinical trials and epidemiological studies. Closer to home, reviews of published studies find a high frequency of poor quality in research methods, including those used for statistical analysis. The statistical literature has long recognized that questionable research findings can occur when investigators fail to set aside their own outcome preferences as they analyse and interpret data. These preferences can be related to financial interests, a concern for patients, peer recognition, and commitment to a hypothesis. Several analyses of published papers provide evidence of an association between financial conflicts of interest and reported results. If we are to regain professional and lay confidence in research findings some changes are required. Clinical journals need to develop more competence in the review of analytic methods and provide space for thorough discussion of published papers whose results are challenged. Graduate schools need to prepare students for the conflicting interests that surround the practice of statistics. Above all, each of us must recognize our responsibility to use analytic procedures that illuminate the research issues rather than those serving special interests. PMID- 14566925 TI - The training of biostatistical scientists. PMID- 14566926 TI - Review of the prevalence and incidence of eating disorders. AB - OBJECTIVE: To review the literature on the incidence and prevalence of eating disorders. METHODS: We searched Medline using several key terms relating to epidemiology and eating disorders and we checked the reference lists of the articles that we found. Special attention has been paid to methodologic problems affecting the selection of populations under study and the identification of cases. RESULTS: An average prevalence rate for anorexia nervosa of 0.3% was found for young females. The prevalence rates for bulimia nervosa were 1% and 0.1% for young women and young men, respectively. The estimated prevalence of binge eating disorder is at least 1%. The incidence of anorexia nervosa is 8 cases per 100,000 population per year and the incidence of bulimia nervosa is 12 cases per 100,000 population per year. The incidence of anorexia nervosa increased over the past century, until the 1970s. DISCUSSION: Only a minority of people who meet stringent diagnostic criteria for eating disorders are seen in mental health care. PMID- 14566928 TI - The place of inpatient care in the treatment of anorexia nervosa: questions to be answered. AB - BACKGROUND: The chance that an anorexia nervosa patient will be hospitalized depends more on circumstantial rather than on scientifically based factors. Although there is a lot of information on the treatment of anorexia nervosa patients in a residential setting, answers to questions relating to the "when," "where," and "how" of treatment are subjective. There is no clinical consensus and the paucity of controlled research is hampering the development of an evidence-based practice. RESULTS: Increasing economic restraints through managed care policies limit the length of inpatient treatment, which leads to early discharge at a lower body weight, which leads to a higher likelihood of readmissions, which leads to increasing costs. DISCUSSION: We will highlight important issues in the ongoing debate between economic demands and clinical challenges. Our goal is to stimulate critical reflections and systematic research. PMID- 14566927 TI - Risk factors for anorexia nervosa: a national cohort study. AB - OBJECTIVE: To identify possible risk factors for anorexia nervosa through national registers. METHOD: The study includes the entire Swedish population of 989,871 individuals born between 1973 and 1982. Patients with anorexia nervosa were identified through the Swedish Hospital Discharge Register from November 1990 to December 1999. Information about sociodemographic, perinatal, and psychosocial variables was obtained from different national registers. Attributable risk (AR) was calculated for potential risk factors. RESULTS: Females had the highest AR for hospital admission because of anorexia nervosa (89.2%). Another important risk factor was having parents born in northern, central, or eastern Europe (AR: 49.3%). Psychosocial risk factors also were associated with an increased risk for anorexia nervosa (AR: 7.6%), whereas perinatal complications had an AR of only 3.6%. DISCUSSION: The most important risk factors were related to the sociocultural context of the individual, thus supporting hypotheses of a sociocultural etiology of anorexia nervosa. PMID- 14566929 TI - Binge eating and binge eating disorder in a small-scale, indigenous society: the view from Fiji. AB - OBJECTIVE: Although the cross-cultural prevalence of anorexia and bulimia nervosa has been investigated in multiple studies, little is known about the prevalence and correlates of binge eating and binge eating disorder (BED) cross-culturally. No published studies to date have explored BED in small-scale, indigenous, or developing societies. The current study investigated the prevalence and correlates of binge eating in a community sample of Fijian women living in rural Fiji. METHODS: Fifty ethnic Fijian women completed a self-report measure developed for this study on dieting and attitudes toward body shape and change, a Nadroga-language questionnaire on body image, and the Questionnaire on Eating and Weight Patterns-Revised (QEWP-R). Their height and weight were also measured. Patterns of dieting, high body mass index (BMI), and attitudes toward eating and body image were compared between women with and without a history of binge eating. RESULTS: Ten percent of respondents reported at least weekly episodes of binge eating during the past 6 months and 4% endorsed symptoms consistent with BED. Binge eating in this sample was associated significantly with a BMI value above 35, a history of dieting, and a high concern with body shape. Binge eating was not associated with several markers of acculturation in this sample, although it was associated with a key, nontraditionally Fijian (i.e., acculturated) attitude toward the body. DISCUSSION: Binge eating occurred in a social context with traditions concerning weight and diet widely disparate from Western populations. However, correlates of binge eating in this sample suggest that nontraditional Fijian attitudes toward weight and body shape play a contributory role. PMID- 14566930 TI - Circulating levels of neuroactive steroids in patients with binge eating disorder: a comparison with nonobese healthy controls and non-binge eating obese subjects. AB - OBJECTIVES: Increased plasma levels of allopregnenolone (3alpha,5alpha tetrahydroprogesterone [3alpha,5alpha-THP]), dehydroepiandrosterone (DHEA), and DHEA sulphate (DHEA-S) have been reported in patients with anorexia nervosa or bulimia nervosa. To assess whether those changes are related to malnutrition, we investigated plasma levels of neuroactive steroids in women with binge eating disorder (BED) who compulsively binge as bulimic patients, but do not incur malnutrition. METHODS: Sixty-eight women participated in the study (31 nonobese healthy controls, 9 nonobese patients with BED, 16 obese patients with BED, and 12 obese non-binge eating women). Blood samples were collected in the morning for determination of plasma levels of 3alpha,5alpha-THP, DHEA, DHEA-S, and cortisol. RESULTS: Nonobese BED women had significantly higher plasma levels of DHEA, DHEA S, and 3alpha,5alpha-THP than nonobese healthy women. Similarly, obese individuals with BED exhibited significantly higher neurosteroid plasma levels than non-binge eating obese subjects. No significant differences in plasma cortisol levels were observed among the groups. DISCUSSION: This study shows increased plasma levels of neuroactive steroids in BED patients. These findings could have been influenced by methodologic limitations (e.g., the absence of diurnal sampling). However, they suggest that if malnutrition is involved in the determination of increased plasma levels of neuroactive steroids in people with anorexia or bulimia nervosa, then different factors may induce similar effects in people with BED. Alternatively, common unknown factors could be responsible for neurosteroid changes in anorexia nervosa, bulimia nervosa, and BED. PMID- 14566931 TI - Relative efficacy of a 4-day versus a 5-day day hospital program. AB - OBJECTIVE: Day hospital treatment has demonstrated effectiveness in the treatment of eating disorders, but no information regarding the required intensity of treatment is currently available. The purpose of this study was to compare the short-term effectiveness of a 4-day versus a 5-day day hospital program. METHODS: The study followed a sequential cohort design. Four hundred sixty-eight patients attended a 5-day program between 1985 and 1994 and 288 patients attended a 4-day program between 1995 and 2000. Weight, binging, and vomiting frequencies and psychological functioning were assessed at the beginning and at the end of treatment. RESULTS: The two programs were equally effective in helping underweight patients to gain weight. However, the 5-day program was associated with higher abstinence rates for binging and vomiting and with better psychological functioning at the end of treatment. The 4-day program was more cost-effective when symptom change was considered as a percentage reduction per treatment day. DISCUSSION: Overall, the 5-day program was both more effective and more costly. The relative long-term outcome of the two programs is a critical question and these data are currently being collected. PMID- 14566932 TI - Family interactions predict intraindividual symptom variation for adolescents with bulimia. AB - OBJECTIVE: Following family systems and stress theories, this study considered predictors of symptom variation experienced by adolescents with bulimia. METHODS: Using experience sampling methodology, 20 girls clinically diagnosed with bulimia nervosa completed questionnaires about hassles and bulimic symptoms for 1 week, eight times daily, whenever they were contacted by pager. Perceived family environment was defined by the Family Environment Scale and the Conflict Behavior Questionnaire. RESULTS: Regression analyses found that potent family hassles were positive predictors of bulimic symptoms later that day for girls who perceived their family as having high levels of conflict or low levels of emotional expressiveness. This was not found when girls perceived their family environment as more normal. DISCUSSION: Within the context of a perceived dysfunctional family environment, potent family hassles can predict interindividual and intraindividual bulimic symptom variation for adolescent girls. PMID- 14566933 TI - Improved DNA flow cytometric, DNA ploidy, and S-phase reproducibility between 15 laboratories in analysis of breast cancer using generalized guidelines. AB - BACKGROUND: Lack of generalized guidelines for DNA flow cytometric analysis (FCM) may be the main reason for its limited use in the clinical management of breast cancer. METHODS: After an initial interlaboratory reproducibility study (Round I), we concluded that it was the evaluation of the DNA histograms rather than the technical performance of the analysis that was the main reason for discordant results between laboratories. Guidelines for the interpretation of DNA histograms were therefore drawn up. We present here data from a new reproducibility study (Round II) using these guidelines. RESULTS: For 10 laboratories also participating in Round I, use of the guidelines increased the concordance in DNA ploidy status from 89% to 100% for the 46 samples used in both rounds. The concordance rate for SPF also increased; mean r(s)-value increased from 0.81 to 0.88, and mean kappa value (lower two-thirds versus upper third versus not reported) increased from 0.55 to 0.71. Five new laboratories, participating only in Round II, also agreed with the 10 original laboratories regarding DNA ploidy status. With the inclusion of all 15 laboratories, we obtained a mean r(s)-value of 0.81 and a mean kappa value of 0.72 for SPF. CONCLUSIONS: Generalized guidelines for DNA FCM increase interlaboratory agreement, which is highly important in clinical routines and in multicenter studies. Furthermore, inexperienced FCM laboratories using generalized guidelines can produce and interpret DNA FCM data equally as well as experienced laboratories. PMID- 14566934 TI - Monitoring cell cycle distributions in living cells by videomicrofluorometry and discriminant factorial analysis. AB - BACKGROUND: The study of the cell cycle of living cells is often based on quantification of nuclear DNA. These studies may be improved by multifactorial analysis evaluating several parameters for each cell. METHODS: Single lymphoblastoid living cells were labeled with three fluorescent markers: Hoechst 33342 for nuclear DNA, Rhodamine 123 for mitochondria, and Nile Red for plasma membrane. Numerical image analysis allowed us to obtain, for each cell, morphological parameters (e.g., cell size, nuclear size, and shape) and functional information (e.g., nuclear DNA content, level of mitochondria energetic state, and the amount and properties of the plasma membrane) by fluorescence intensity. These parameters were used in a typological analysis that separated control cells into four groups. RESULTS: A discriminant factorial analysis (DFA) confirmed the four groups: G0-G1, S, G2+M, and polyploid cells called Gn. These groups were significantly different, with a classification probability of 0.9999; these control cells defined a learning population. Different populations of untreated and adriamycin-treated cells were analyzed as additional individuals within a DFA and were classified into the G0-G1, S, G2+M, and Gn groups by their probability of belonging to each of the groups. CONCLUSIONS: This approach is particularly efficient when it is used to determine variations in cellular properties and to objectively study cellular populations. PMID- 14566935 TI - Flow cytometry and GFP: a novel assay for measuring the import and turnover of nuclear-encoded mitochondrial proteins in live PC12 cells. AB - BACKGROUND: Mitochondrial protein import is typically measured by adding radiolabeled precursor proteins to isolated mitochondria. We have developed a novel, high-throughput method for measuring protein import in live differentiated PC12 cells using a tetracycline (Tet) regulated, nuclear encoded, mitochondrially targeted GFP fusion protein and flow cytometry. METHODS: We generated a PC12 cell line stably transfected with an inducible GFP fusion protein (GFPmt) targeted to mitochondria. GFPmt PC12 cells were treated with NGF for one week to induce neuronal differentiation in the presence of Tet to silence GFP expression. On day seven GFPmt expression was induced by removal of Tet and these "GFP-on" cells were exposed to sublethal levels of CCCP (2 microM) for 24 h. At 24 h, the cells were harvested in Ca(++)-free PBS and the GFPmt signal in live intact cells was measured using flow cytometry. Since GFPmt is not fluorescent prior to being imported into mitochondria, the GFPmt signal reflected only GFPmt imported to mitochondria. PI was used to gate out contributions from dead cells. Turnover of GFPmt in mitochondria was also assessed; in this case, Tet was added to arrest GFPmt expression in GFP-on cells, and the subsequent decline of the fluorescent signal, in the absence of any new GFP synthesis, was measured by flow cytometry. RESULTS: Exposure to 2 microM CCCP for 24 h caused a 61% +/- 0.4 decline in GFPmt fluorescence compared to controls. This decline corresponded to a 30% +/- 7 decrease in GFPmt protein levels measured by Western blot of mitochondrial fractions, and a 72% +/- 5 decline in the import of newly synthesized GFPmt to mitochondria over a 1 h period 24-h after addition of 2 microM CCCP measured by autoradiography. CCCP partially depolarized mitochondria but was not lethal for up to five days. CONCLUSIONS: This novel GFP-based flow cytometry assay is a rapid and sensitive technique for quantifying protein import to mitochondria in live neuronal cells. PMID- 14566936 TI - A hybrid 3D watershed algorithm incorporating gradient cues and object models for automatic segmentation of nuclei in confocal image stacks. AB - BACKGROUND: Automated segmentation of fluorescently-labeled cell nuclei in 3D confocal microscope images is essential to many studies involving morphological and functional analysis. A common source of segmentation error is tight clustering of nuclei. There is a compelling need to minimize these errors for constructing highly automated scoring systems. METHODS: A combination of two approaches is presented. First, an improved distance transform combining intensity gradients and geometric distance is used for the watershed step. Second, an explicit mathematical model for the anatomic characteristics of cell nuclei such as size and shape measures is incorporated. This model is constructed automatically from the data. Deliberate initial over-segmentation of the image data is performed, followed by statistical model-based merging. A confidence score is computed for each detected nucleus, measuring how well the nucleus fits the model. This is used in combination with the intensity gradient to control the merge decisions. RESULTS: Experimental validation on a set of rodent brain cell images showed 97% concordance with the human observer and significant improvement over prior methods. CONCLUSIONS: Combining a gradient-weighted distance transform with a richer morphometric model significantly improves the accuracy of automated segmentation and FISH analysis. PMID- 14566937 TI - A simple way of quantifying immunostained cell nuclei on the whole histologic section. AB - BACKGROUND: Quantification of nuclei, immunostained for proliferation or differentiation markers, is widely recommended for prognostication and treatment of numerous solid tumors. However, many problems concerning reproducibility of the results obtained still remain, partially due to the lack of standardization of the method used and to the difficulty of taking into account tumor tissue heterogeneity. Technical solutions exist but they are expensive and time consuming, and their complexity limits their diffusion in routine pathology departments. METHODS: In the present study, we use a simple, fast and inexpensive system of acquisition, a 4,000 dots-per-inch slide scanner, to get a single true color numerical image of the whole histologic section. The analysis is done thanks to fully automatic image processing, allowing computation of the stain proportion on the whole histologic section, as well as in "hot spots." RESULTS: After having precisely fixed settings of the slide scanner, one can easily detect counterstained and immunostained structures, by image analysis. This simple and stable processing can be done under 6 min. CONCLUSIONS: The association of immunohistochemistry, stable acquisition device and computer-assisted image analysis provides an objective, reproducible, and easy to standardize quantification of the nuclear markers, in relation to the total tumor tissue architecture. PMID- 14566938 TI - Genetic heterogeneity and ploidy level analysis among different gynogenetic clones of the polyploid gibel carp. AB - BACKGROUND: Some triploid and tetraploid clones have been identified in the gynogenetic gibel carp, Carassius auratus gibelio Bloch, by karyotypic and cytologic analyses over many years. Further, 5-20% males and karyotypic diversity have been found among their natural and artificial populations. However, the DNA contents and the relation to their ploidy level and chromosome numbers have not been ascertained, and whether normal meiosis occurs in spermatogenesis needs to be determined in the different clones. METHODS: The sampled blood cells or sperms were mixed with blood cells from chicken or individual gibel carp and fixed in 70% pre-cooled ethanol overnight at 4 degrees C. The mixed cell pellets were washed 2-3 times in 1x phosphate buffered saline and then resuspended in the solution containing 0.5% pepsin and 0.1 M HCl. DNA was stained with propidium iodide solution (40 microg/mL) containing 4 kU/ml RNase. The measurements of DNA contents were performed with Phoenix Flow Systems. RESULTS: Triploid clones A, E, F, and P had almost equal DNA content, but triploid clone D had greater DNA content than did the other four triploid clones. DNA content of clone M (7.01 +/- 0.15 pg/nucleus) was almost equal to the DNA content of clone D (5.38 +/- 0.06 pg/nucleus) plus the DNA content of common carp sperm (1.64 +/- 0.02 pg/nucleus). The DNA contents of sperms from clones A, P, and D were half of their blood cells, suggesting that normal meiosis occurs in spermatogenesis. CONCLUSIONS: Flow cytometry is a powerful method to analyze genetic heterogeneity and ploidy level among different gynogenetic clones of polyploid gibel carp. Through this study, four questions have been answered. (a) The DNA content correlation among the five triploid clones and one multiple tetraploid clone was revealed in the gibel carp, and the contents increased with not only the ploidy level but also the chromosome number. (b) Mean DNA content was 0.052 pg in six extra chromosomes of clone D, which was higher than that of each chromosome in clones A, E, F, and P (about 0.032 pg/chromosome). This means that the six extra chromosomes are larger chromosomes. (c) Normal meiosis occurred during spermatogenesis of the gibel carp, because DNA contents of the sperms from clones A, P, and D were almost half of that in their blood cells. (d) Multiple tetraploid clone M (7.01 +/- 0.15 pg/nucleus) contained the complete genome of clone D (5.38 +/- 0.06 pg/nucleus) and the genome of common carp sperm (1.64 +/- 0.02 pg/nucleus). PMID- 14566939 TI - Cell-mediated cytotoxicity evaluation using monoclonal antibody staining for target or effector cells with annexinV/propidium iodide colabeling by fluorosphere-adjusted counts on three-color flow cytometry. AB - BACKGROUND: In addition to (51)chromium release assay, flow cytometric methods have been described to assess in vitro cell-mediated cytotoxicity. In this report, we describe a new flow cytometric approach for determination of in vitro cell-mediated cytotoxicity utilizing three-color flow cytometric assay. METHODS: This method is based on monoclonal antibody staining of either effector or target cells to evaluate cytotoxicity with increased accuracy by utilizing fluorospheres for calibration. The basic strategy involves labeling effector or target cells with a specific fluorescent-conjugated monoclonal antibody, in addition to staining with annexinV-FITC and propidium iodide to identify apoptotic/dead cells. The effector and target cell populations as well as conjugates were clearly and easily identified by this approach. RESULTS: We obtained significant correlation between cytotoxicity calculated by this technique and (51)chromium release assay results. The integration of fluorospheres allowed us to determine the absolute number of events reflective of the cumulative cell death rather than a cross-sectional, percentage-based cytotoxicity assessment in the target cell population at the time of analysis. CONCLUSIONS: This method provides additional advantages to other methods and enables the study of target cell fate in more detail, as well as providing a potential contribution to understanding the mechanisms of cell elimination. PMID- 14566941 TI - Cellular interactions between axon terminals containing endogenous opioid peptides or corticotropin-releasing factor in the rat locus coeruleus and surrounding dorsal pontine tegmentum. AB - Recent evidence suggests that certain stressors release both endogenous opioids and corticotropin-releasing factor (CRF) to modulate activity of the locus coeruleus (LC)-norepinephrine (NE) system. In ultrastructural studies, axon terminals containing methionine(5)-enkephalin (ENK) or CRF have been shown to target LC dendrites. These findings suggested the hypothesis that both neuropeptides may coexist in common axon terminals that are positioned to have an impact on the LC. This possibility was examined by using immunofluorescence and immunoelectron microscopic analysis of the rat LC and neighboring dorsal pontine tegmentum. Ultrastructural analysis indicated that CRF- and ENK-containing axon terminals were abundant in similar portions of the neuropil and that approximately 16% of the axon terminals containing ENK were also immunoreactive for CRF. Dually labeled terminals were more frequently encountered in the "core" of the LC vs. its extranuclear dendritic zone, which included the medial parabrachial nucleus (mPB). Triple labeling for ENK, CRF, and tyrosine hydroxylase (TH) showed convergence of opioid and CRF axon terminals with noradrenergic dendrites as well as evidence for inputs to TH-labeled dendrites from dually labeled opioid/CRF axon terminals. One potential source of ENK and CRF in the dorsal pons is the central nucleus of the amygdala (CNA). To determine the relative contribution of ENK and CRF terminals from the CNA, the CNA was electrolytically lesioned. Light-level densitometry revealed robust decreases in CRF immunoreactivity in the LC and mPB on the side ipsilateral to the lesion but little or no change in ENK immunoreactivity, confirming previous studies of the mPB. Degenerating terminals from the CNA in lesioned rats were found to be in direct contact with TH-labeled dendrites. Together, these data indicate that ENK and CRF may be colocalized to a subset of individual axon terminals in the LC "core." The finding that the CNA provides, to dendrites in the area examined, a robust CRF innervation, but little or no opioid innervation, suggests that ENK and CRF axon terminals impacting LC neurons originate from distinct sources and that terminals that colocalize ENK and CRF are not from the CNA. PMID- 14566942 TI - Fine structural localization of connexin-36 immunoreactivity in mouse cerebral cortex and thalamus. AB - The mounting physiological evidence for low-resistance electrical coupling between thalamic and cortical neurons contrasts with a lack of morphological data on gap junctions in thalamus and cortex. Connexin-36 is a neuronally specific protein associated with low-resistance gap junctions in the central nervous system. Connexin-36 localization was studied in the mouse somatosensory cortex and thalamus by using immunocytochemistry and immunoelectron microscopy. Expression of connexin-36 immunoreactivity is widespread in the forebrain and significantly enhanced in the barrel cortex and thalamic reticular nucleus during the second postnatal week, but it extends to other thalamic nuclei as well. At the electron microscopic level, pre- and postembedding immunogold labeling revealed that 70-76% of connexin-36-immunolabeled particles were localized at focal sites on apposed plasma membranes of cortical and thalamic dendrites; approximately 5% of the particles were associated with parasynaptic membranes; but on no occasion could overt, morphologically identifiable gap junctions be demonstrated in association with connexin-36 immunoreactivity. The widespread distribution of focal concentrations of connexin-36 subunits could provide a basis for the electrical coupling that exists between cortical and reticular thalamic neurons, but morphologically definable gap junctions may be too small to be adequately visualized by conventional immunoelectron microscopy. PMID- 14566943 TI - Contribution of connexin26 to electrical feedback inhibition in the turtle retina. AB - The first synaptic integration in the neuronal cascade of vision in vertebrates includes a feedback from horizontal cells to cones by a mechanism yet not fully understood. Recent observations in teleosts suggested an electrical feedback mechanism mediated by connexin26 (Cx26) hemichannels at the terminal dendrites of horizontal cells. By using reverse transcription-polymerase chain reaction and immunoblotting from retinal homogenate, we detected Cx26 mRNA transcripts in the turtle retina and demonstrated that they were translated into protein. Cx26 immunoreactivity was especially prominent in the outer plexiform layer. Subcellularly, immunoreactivity was located mainly between horizontal cell axon terminals and in horizontal cell dendrites forming the lateral elements at the ribbon synaptic complex of the cone pedicle. The label was confined to the horizontal cell membrane flanking the ribbon and was not found on the opposing photoreceptor membrane. No gap junctions at this location are known, so immunosignaling suggested the presence of hemichannels. Their relevance to the feedback mechanism was investigated by intracellular recordings from horizontal cells during application of the hemichannel blocker carbenoxolone. Carbenoxolone hyperpolarized the dark membrane potential by about 25 mV, decreased the amplitudes of responses to full-field light flashes, and suppressed the feedback induced depolarizing inflexion in the response profile. These physiological findings are compatible with the involvement of hemichannels in the feedback between horizontal cells and cones and support the anatomical findings. Together, these data suggest the presence of an electrical feedback mechanism in the turtle retina, which therefore might be a common mechanism at the first visual synapse in vertebrates. PMID- 14566944 TI - Ultrastructural localization of the norepinephrine transporter in superficial and deep layers of the rat prelimbic prefrontal cortex and its spatial relationship to probable dopamine terminals. AB - The prefrontal cortex (PFC) is a likely site of action for the therapeutic efficacy of antidepressants that inhibit norepinephrine (NE) reuptake. Moreover, drugs that block the NE transporter (NET) increase extracellular levels of both NE and dopamine (DA), an interaction that may contribute to their therapeutic properties. To examine the subcellular localization of NET and to investigate the spatial relationships between presumed NE and DA axons within the rat prelimbic PFC, we combined immunogold-silver localization of NET with immunoperoxidase staining for the catecholamine synthetic enzyme tyrosine hydroxylase (TH). An additional aim was to quantify the proportion of profiles dually labeled for NET and TH to test the common observation that TH immunolabeling is relatively selective for DA axons. NET-immunoreactive (NET-ir) axonal profiles were typically unmyelinated and occasionally were observed to form symmetric axodendritic synapses. The majority of immunogold NET labeling was unexpectedly observed in the cytoplasm rather than on the plasma membrane. Furthermore, in tissue dually labeled for both NET and TH, only 8-10% of profiles contained both markers. Unlike observations for singly labeled profiles, gold-silver particles for NET in dually labeled axons were localized primarily to the plasmalemma. A systematic survey of terminals labeled only for TH revealed that they were typically separated by at least 1.2 mum from NET-ir varicosities, and the two profile types were not seen to contact common targets. These results suggest that, in the rat PFC, NE axons (1) contain predominantly cytoplasmic NET, (2) infrequently contain TH immunolabeling, and (3) may interact with probable DA afferents by means of extrasynaptic mechanisms. PMID- 14566945 TI - Differential expression of nitric oxide in serotonergic projection neurons: neurochemical identification of dorsal raphe inputs to rodent trigeminal somatosensory targets. AB - The dorsal raphe (DR) is invested with nitric oxide synthase (NOS)-expressing profiles. To characterize the connections of NO-containing cells and further assess neurochemical relationships maintained by DR, the transmitter identity of the raphe projection to the trigeminal somatosensory system was examined. Rats were injected with retrograde tracer into vibrissae-related target areas or with anterograde tracer into DR. NADPH-diaphorase (NADPHd) histochemistry or NOS immunostaining was combined with serotonin (5HT)- or serotonin transporter (SERT) immunolabeling to examine: 1) the presence of NO in 5HT-containing axons from DR; 2) the distribution of NO-containing fibers with respect to other nitrergic profiles in the somatosensory system; and 3) the propensity for individual projection neurons in specific subdivisions of DR to colocalize 5HT and NO. Results confirm that "barrel-like" patches can be identified in several adult trigeminal relay nuclei by NADPHd histochemistry and demonstrate that fibers from DR contain 5HT and NO. Observations include a high percentage of cortical midline projection neurons which contained NADPHd (70-80%) and coexpressed 5HT. In contrast, approximately 40% of retrogradely labeled DR-thalamus cells in the lateral wing demonstrated NADPHd or 5HT expression, but not both in the same neuron. Colocalization of NADPHd and 5HT within individual DR projection neurons indicates that: i) DR is a source of nitrergic input to trigeminal structures, and ii) NO and 5HT may be simultaneously released to influence information processing within somatosensory targets. Disparities in NADPHd expression between retrogradely labeled DR neuronal subpopulations further suggest functional differences in the impact of NO on cortical and subcortical targets. PMID- 14566946 TI - Crossed and uncrossed retinal projections to the hamster circadian system. AB - The hamster suprachiasmatic nucleus (SCN), site of the circadian clock, has been thought to be equally and completely innervated by each retina. This issue was studied in animals that had received an injection of the tracer cholera toxin subunit B (CTb) conjugated to Alexa 488 into the vitreous of one eye, with CTb Alexa 594 injected into the other. Retinal projections to the SCN and other nuclei of the circadian system were simultaneously evaluated by using confocal laser microscopy. Each retina provides completely overlapping terminal fields throughout each SCN. Although SCN innervation by the contralateral retina is slightly denser than that from the ipsilateral retina, there are distinct SCN regions where input from one side is predominant, but not exclusive. A dense terminal field from the contralateral retina encompasses, and extends dorsally beyond, the central SCN subnucleus identified by calbindin-immunoreactive neurons. Surrounding the dense terminal field, innervation is largely derived from the ipsilateral retina. The densest terminal field in the intergeniculate leaflet is from the contralateral retina, which completely overlaps the ipsilateral projection. Most nuclei of the pretectum receive innervation largely, but not solely, from the contralateral retina, although the olivary pretectal nucleus has very dense patches of innervation derived exclusively from one retina or the other. Retina-dependent variation in terminal field density within the three closely examined nuclei may indicate areas of specialized function not previously appreciated. This issue is discussed in the context of the melanopsin containing retinal ganglion cell projections to several nuclei in the circadian visual system. PMID- 14566947 TI - Sensorimotor corticocortical projections from rat barrel cortex have an anisotropic organization that facilitates integration of inputs from whiskers in the same row. AB - We used a dual anterograde-tracing paradigm to characterize the organization of corticocortical projections from primary somatosensory (SI) barrel cortex. In one group of rats, biotinylated dextran amine (BDA) and Fluoro-Ruby (FR) were injected into separate barrel columns that occupied the same row of barrel cortex; in the other group, the tracers were deposited into barrel columns residing in different rows. The labeled corticocortical terminals in the primary motor (MI) and secondary somatosensory (SII) cortices were plotted, and digital reconstructions of these plots were quantitatively analyzed. In all cases, labeled projections from focal tracer deposits in SI barrel cortex terminated in elongated, row-like strips of cortex that corresponded to the whisker representations of the MI or SII cortical areas. When both tracers were injected into separate parts of the same SI barrel row, FR- and BDA-labeled terminals tended to merge into a single strip of labeled MI or SII cortex. By comparison, when the tracers were placed in different SI barrel rows, both MI and SII contained at least two row-like FR- and BDA-labeled strips that formed mirror image representations of the SI injection sites. Quantitative analysis of these labeling patterns revealed three major findings. First, labeled overlap in SII was significantly greater for projections from the same barrel row than for projections from different barrel rows. Second, in the infragranular layers of MI but not in the supragranular layers, labeled overlap was significantly higher for projections from the same SI barrel row. Finally, in all layers of SII and in the infragranular layers of MI, the amount of labeled overlap was proportional to the proximity of the tracer injection sites. These results indicate that SI projections to MI and SII have an anisotropic organization that facilitates the integration of sensory information received from neighboring barrels that represent whiskers in the same row. PMID- 14566948 TI - Paleocortex is specified in mice in which dorsal telencephalic patterning is severely disrupted. AB - The patterning of the telencephalon is regulated by the concerted action of distinct mechanisms operating in different portions of this structure. Although much progress has been made in understanding the mechanisms underlying the specification of dorsal and ventral structures, little is known about the specification of the paleocortex, the olfactory cortex located at the interface of the dorsal and ventral telencephalon. The paleocortex is thought to be a dorsal, cortical structure, derived from the lateral extreme of the dorsal telencephalon. We examined mutant mice in which dorsal telencephalic patterning is severely disrupted, to ask how these perturbations affect the paleocortex. In the Lhx2-/- telencephalon, where the cortex is greatly shrunken such that medial and dorsal cortical tissue is undetectable, normal expression of several paleocortical markers is observed. The Gli3-/- telencephalon, where the dorsal telencephalon is ventralized, also displays paleocortical markers. In contrast, when the ventral telencephalon is almost completely deleted, such as in the BF1-/ brain, paleocortical markers are undetectable. These results indicate that the specification of the paleocortex can occur in spite of drastic perturbations of dorsal patterning. Furthermore, in the Lhx2 mutant, the paleocortex is juxtaposed to an expanded and mislocated source of Wnt and Bmp signaling, the cortical hem, whereas, in the Gli3 mutant, paleocortical markers arise even though the cortical hem is missing. This indicates that an increase or decrease in cues from this dorsal signaling center does not disrupt the specification of the paleocortex. Finally, by using an in vitro assay, we found that isolated explants of lateral telencephalon up-regulate normal expression of paleocortical markers when maintained in vitro, from as early as embryonic day (E) 10.5. Together, the results reveal that, although the paleocortex is considered to be a cortical structure, it is specified even when dorsal telencephalic patterning is grossly perturbed. Furthermore, our in vitro data reveal that, if mechanisms outside the lateral telencephalon are involved in the specification of the paleocortex, they must act extremely early, prior to E10.5. PMID- 14566949 TI - Stereotyped neuropil branching of an identified stomatogastric motor neuron. AB - Anatomical studies of the crab stomatogastric ganglion (STG) have suggested only minimal organization within the neuropil of this structure. Here, we present evidence that, for at least one intrinsic neuron type, the ventricular dilator (VD) neuron, a highly organized and stereotyped branching structure exists within the stomatogastric neuropil. Specifically, we show the morphology of the VD neuron consists of a single primary neurite that projects from the soma into the neuropil and bifurcates into a pair of subprimary neurites, which in turn exit the neuropilar region, one entering the left and the other the right medial ventricular nerve. Nearly all secondary neurite branching of the VD neuron is from the subprimary neurites. There are approximately 22 secondary branches/neuron (range 14-28), with no significant difference between the number of secondary branches off the right vs. the left subprimary neurite, although the ratio of secondary branches between subprimaries varies (range 0.4-1.6). The fine neurites that branch from the secondary processes segregate hemispherically within the neuropil, based on the subprimary neurite of origin. Within this hemispherical organization, another level of fine neurite segregation is present, namely, the fine neurites derived from each secondary branch are restricted to discrete regions of the hemisphere with only minimal overlap with those derived from other secondary branches. Monte Carlo simulations show that this segregation differs significantly from a random distribution. The organization of branching seen in the VD neuron may play a critical role in the electrotonic and local computational organization of this neuron and sets the stage for physiological experimentation addressing these issues. PMID- 14566950 TI - Extraordinary diversity in vasopressin (V1a) receptor distributions among wild prairie voles (Microtus ochrogaster): patterns of variation and covariation. AB - The vasopressin V1a receptor is a gene known to be central to species differences in social behavior, including differences between the monogamous prairie vole and its promiscuous congeners. To examine how individual differences compare with species differences, we characterize variability in the expression of the vasopressin V1a receptor (V1aR) in a large sample of wild prairie voles. We find a surprising degree of intraspecific variation in V1aR binding that does not seem attributable to experimental sources. Most brain regions exhibit differences between upper and lower quartiles that are comparable to differences between species in this genus. Regions that are less variable have been implicated previously in regulating monogamous behaviors, suggesting that the lack of variation at these sites could reflect natural selection on mating system. Many brain regions covary strongly. The overall pattern of covariation reflects the developmental origins of brain regions. This finding suggests that shared mechanisms of transcriptional regulation may limit the patterns of gene expression. Such biases may shape both the efficacy of selection and the pattern of individual and species differences. Overall, our data indicate that the prairie vole would be a useful model for exploring how individual differences in gene expression influence complex social behaviors. PMID- 14566952 TI - Adolescent psychotherapy: an introduction. AB - Adolescent psychotherapy has only recently taken hold as distinct from either child or adult psychotherapy. This new recognition was born of necessity as practitioners saw that theories and techniques developed for treating children and adults were far less effective for treating adolescents. As necessity is often the mother of invention, new models for treating adolescents have emerged. While these models differ in many respects, they share three characteristics: the unique features of adolescents, psychotherapists requiring specialized knowledge and skills, and the need for adolescent psychotherapy to involve the broader systemic context of the adolescent's life. The articles in this issue of In Session: Journal of Clinical Psychology insist that practitioners recognize and challenge their assumptions about how psychotherapy with adolescents should be conducted. The articles highlight the need for specialized training, the scarcity of evidenced-based research, and the ethical dilemmas that frequently occur when working with this population. PMID- 14566951 TI - Timing and topography of nucleus magnocellularis innervation by the cochlear ganglion. AB - This series of experiments examined the arrival and organization of cochlear nerve axons in the primary auditory brainstem nucleus, nucleus magnocellularis (NM), of the chick. DiI and DiD were injected into the cochlear nerve, cochlear ganglion, and basilar papilla (i.e., avian cochlea) in fixed tissue and labeled axons were studied in NM and its vicinity. Cochlear nerve axons first penetrate NM between stages 29 (E6) and 36 (E10). Axons penetrate NM in a middle-to posterior-to-anterior developmental sequence; the anterior, high-frequency region of NM receives axons last. When cochlear nerve axons arrive in the NM, they are already organized in a topographic map related to the position of their cell bodies along the basilar papilla, foreshadowing the tonotopic mapping observed between NM and the basilar papilla later in development. Evidence of a topographic map was also observed in the other primary auditory brainstem nucleus, nucleus angularis. These results indicate that topographic mapping of position (and ultimately characteristic frequency) between the basilar papilla and NM is established as cochlear nerve axons arrive in the NM prior to the onset of synaptic activity. . PMID- 14566953 TI - The outcomes of psychotherapy with adolescents: a practitioner-friendly research review. AB - This article summarizes the outcome findings of psychotherapy with adolescents through an examination of the research literature. In this problem-focused review of the research literature, we consider adolescent treatment of anxiety disorders, depression, eating disorders, disruptive behavior disorders, multiple problems of adolescence, and health risk behaviors. The evidence base for the effectiveness of various psychotherapies for adolescents is mixed. Psychotherapy in general has been a ripe field in which unsupported approaches have grown, and this is certainly true for psychotherapeutic interventions with adolescents. Nonetheless, there is a growing body of evidence to support various types of interventions as effective for different presenting problems. Researchers and clinicians need to recognize the complexity of the diverse characteristics of adolescents, including variations of culture, race, ethnicity, gender, class, physical disability, family definitions and constellation, and sexual orientation. These require sensitivity, responsiveness, and competence by the therapist, but clinical research has not fully examined these considerations. PMID- 14566954 TI - On treating adolescent girls: focus on strengths and resiliency in psychotherapy. AB - This strength-based psychotherapy with adolescent girls and their families is derived from feminist psychology, positive psychology, and strength-based interventions with teens. Research reviewed by the American Psychological Association's Presidential Task Force on Adolescent Girls formed the basis of specific interventions within this approach. Research findings that contributed are the effects on teen girls of positive parental relationships; utilizing strengths of their race, ethnicity, class, and gender; positive body images; being outspoken in relationships; problem-solving skills that foster independence; and family support for independence. The strength-based approach is illustrated by a case example of a 13-year-old European American girl with acting out behaviors, depression, and subclinical attention-deficit/hyperactivity disorder. The case illustrates how to empower adolescent girls within therapy, when and how to include parents, how to change the focus to strengths, and how to help the parents assess and build upon their daughters' strengths. PMID- 14566955 TI - Relational psychoanalytic treatment for young adult males. AB - The clinical treatment of an adolescent young adult male is presented to both illustrate and operationalize the author's hypothesis that a new model of relational psychoanalytic treatment is likely to have beneficial effects, particularly among this population. The contention that males as young boys undergo a normative trauma of premature separation in our society, which is likely to give rise to underdiagnosed abandonment depressive syndromes in teenage boys, is argued. It is represented that psychotherapists attempting to diagnose and treat a wide rage of disorders in young males would do well to rethink their approach from this new perspective. PMID- 14566956 TI - Empowerment psychotherapy with adolescent females of color. AB - We present an empowerment model that fosters strengths and promotes positive identity development among adolescent females of color. Race, ethnicity, gender, and socioeconomic status are integrated in an African American context. A case study illustrates a strength-based approach to conceptualizing and treating a young, Black adolescent female in her cultural context. We conclude with clinical implications for using an empowerment-based approach in psychotherapy with culturally diverse adolescent females. PMID- 14566957 TI - Transforming psychotherapy in order to succeed with adolescent boys: male friendly practices. AB - The purpose of this article is to describe a "male-friendly" therapeutic process with troubled adolescent boys. A male-friendly process is one in which the therapist employs a wide variety of strategies and activities that are likely to appeal to male youth and that facilitate the establishment and maintenance of rapport. The nuances of practicing a male-friendly approach are highlighted and then applied in a case study of psychotherapy with a depressed adolescent boy. The implications of this approach for redefining professional conceptions about boundaries in the client-therapist relationship and reexamining inaccurate stereotypes about the emotional lives of boys are discussed. PMID- 14566958 TI - Clinical application of the gender role strain paradigm: group treatment for adolescent boys. AB - The aim of this article is to demonstrate the clinical application of the gender role strain paradigm with a group of adolescent boys. Adolescent boys experience overwhelming pressure to adhere to gender norms. As they attempt to conform to the expectations associated with the male role, boys suffer from gender role strain. We illustrate how the protective environment of the group enables boys to engage and discuss problems with male role norms. The shift in the boys' adherence to traditional notions of masculinity supports the utility of the gender role strain paradigm in clinical work. PMID- 14566959 TI - Ethical issues in psychotherapy with adolescents. AB - Treating adolescents in psychotherapy presents a number of unique ethical challenges. Because many adolescents who enter treatment have not yet attained the age of majority, reside in families that include other people with emotional difficulties, attend school, become involved with community agencies (e.g., the courts), and must generally take direction from adult authority figures, the role of the therapist becomes particularly complex. The unique dilemmas include developing specialized clinical competence, treatment contracting, choice of treatment modality, direction of the therapeutic process, and confidentiality. This article discusses the range of ethical concerns involved in treating adolescents and suggests strategies for optimal ethical care. PMID- 14566960 TI - Grb10 adapter protein as regulator of insulin-like growth factor receptor signaling. AB - Grb10 is a member of a superfamily of adapter proteins that includes Grb10, 7, 14, and a protein of Caenorhabditis elegans called Mig10. Grb10 proteins are binding partners for several trans-membrane tyrosine-kinase receptors, including the insulin-like growth factor receptor (IGF-IR) and the insulin receptor (IR). Many recent reports have suggested a very important role of Grb10 in regulating IGF-IR signaling. In this review, we will focus on the role of Grb10 in IGF-I induced mitogenesis and we will discuss the recent findings that show the involvement of Grb10 in the regulation of ligand-induced ubiquitination, internalization, and stability of the IGF-IR. PMID- 14566961 TI - How to overcome (and exploit) tumor hypoxia for targeted gene therapy. AB - Tumor hypoxia has long been recognized as a critical issue in oncology. Resistance of hypoxic areas has been shown to affect treatment outcome after radiation, chemotherapy, and surgery in a number of tumor sites. Two main strategies to overcome tumor hypoxia are to increase the delivery of oxygen (or oxygen-mimetic drugs), and exploiting this unique environmental condition of solid tumors for targeted therapy. The first strategy includes hyperbaric oxygen breathing, the administration of carbogen and nicotinamide, and the delivery of chemical radiosensitizers. In contrast, bioreductive drugs and hypoxia-targeted suicide gene therapy aim at activating cytotoxic agents at the tumor site, while sparing normal tissue from damage. The cellular machinery responds to hypoxia by activating the expression of genes involved in angiogenesis, anaerobic metabolism, vascular permeability, and inflammation. In most cases, transcription is initiated by the binding of the transcription factor hypoxia-inducible factor (HIF) to hypoxia responsive elements (HREs). Hypoxia-targeting for gene therapy has been achieved by utilizing promoters containing HREs, to induce selective and efficient transgene activation at the tumor site. Hypoxia-targeted delivery and prodrug activation may add additional levels of selectivity to the treatment. In this article, the latest developments of cancer gene therapy of the hypoxic environment are discussed, with particular attention to combined protocols with ionizing radiation. Ultimately, it is proposed that by adopting specific transgene activation and molecular amplification systems, resistant hypoxic tumor tissues may be effectively targeted with gene therapy. PMID- 14566962 TI - Modulation of vascular smooth muscle cell growth by magnesium-role of mitogen activated protein kinases. AB - We tested the hypothesis that Mg(2+) influences growth of vascular smooth muscle cells (VSMCs) by modulating cell cycle activation through mitogen-activated protein (MAP) kinase-dependent pathways. Rat VSMCs were grown in culture medium containing normal Mg(2+) (1.02 mmol/L, control) and increasing concentrations of Mg(2+) (2-4 mmol/L) for 1-8 days. Effects of varying extracellular Mg(2+) concentration ([Mg(2+)](e)) on intracellular free Mg(2+) concentration ([Mg(2+)](i)) were assessed using mag-fura. Growth actions of Mg(2+) were evaluated by measuring cell cycle activation, DNA synthesis, and protein synthesis. Expression of cell cycle promoters, cyclin D1, cyclin E, Cdk2, and Cdk4 was assessed by immunoblotting. Phosphorylation of cell cycle inhibitors p21(cip1) and p27(kip1) and MAP kinases, ERK1/2, p38MAP kinase, and JNK was evaluated using phospho-specific antibodies. [Mg(2+)](i) increased in a dose dependent manner in response to increasing [Mg(2+)](e). These effects were evident within 2 days and maximal responses were obtained after 6 days. High [Mg(2+)](e) induced cell cycle activation with a lower proportion of cells in G(1) phase (75 +/- 1.0%) and a higher fraction of cells in S phase (12 +/- 0.7%) versus control (G(1), 88.5 +/- 1.4%; S, 6.8 +/- 1.2%; P < 0.05). This was associated with increased protein content of cyclin D1 and Cdk4 and decreased activation of p21(cip1) and p27(kip1). In cells exposed to 2 mmol/L Mg(2+), DNA and protein synthesis was increased approximately threefold. Phosphorylation of MEK1/2 and ERK1/2 was enhanced two to threefold in cells grown in 2 mmol/L Mg(2+). These effects were rapid, occurring within 2 days. Phosphorylation of MEK3/6, p38 MAP kinase, and JNK was unaltered by increasing [Mg2](e). PD98059 (10(-5) mol/L), specific MEK1/2 inhibitor, but not SB202190 (10(-5) mol/L) (specific p38 MAP kinase inhibitor), attenuated Mg(2+)-induced growth actions. These data demonstrate the novel findings that cell cycle activation and growth regulation by Mg(2+) occurs via ERK1/2-dependent, p38 MAP kinase-independent pathways. PMID- 14566963 TI - Butyrate-treated colonic Caco-2 cells exhibit defective integrin-mediated signaling together with increased apoptosis and differentiation. AB - We previously reported that the enterocytic differentiation of human colonic Caco 2 cells correlated with alterations in integrin signaling. We now investigated whether differentiation and apoptosis of Caco-2 cells induced by the short-chain fatty acid butyrate (NaBT) was associated with alterations in the integrin mediated signaling pathway with special interest in the expression and activity of focal adhesion kinase (FAK), of the downstream phosphatidylinositol 3'-kinase (PI 3-kinase)-Akt pathway and in the role of the nuclear factor kappaB (NF kappaB). NaBT increased the level of sucrase. It induced apoptosis as shown by: (1) decreased Bcl-2 and Bcl-X(L) proteins and increased Bax protein; (2) activation of caspase-3; and (3) increased shedding of apoptotic cells in the medium. This effect was associated with defective integrin-mediated signaling as shown by: (1) down-regulation of beta1 integrin expression; 2) decreased FAK expression and tyrosine phosphorylation; (3) concerted alterations in cytoskeletal and structural focal adhesions proteins (talin, ezrin); and (4) decreased FAK ability to associate with PI 3-kinase. However, in Caco-2 cells, beta1-mediated signaling failed to be activated downstream of FAK and PI 3-kinase at the level of Akt. Transfection studies show that NaBT treatment of Caco-2 cells promoted a significant activation of the NF-kappaB which was probably involved in the NaBT-induced apoptosis. Our results indicate that the prodifferentiating agent NaBT induced apoptosis of Caco-2 cells probably through NF-kappaB activation together with a defective beta1 integrin-FAK-PI 3-kinase pathways signaling. PMID- 14566964 TI - Contribution of membrane-associated prostaglandin E2 synthase to bone resorption. AB - This study initially confirmed that, among prostaglandins (PGs) produced in bone, only PGE(2) has the potency to stimulate osteoclastogenesis and bone resorption in the mouse coculture system of osteoblasts and bone marrow cells. For the PGE(2) biosynthesis two isoforms of the terminal and specific enzymes, membrane associated PGE(2) synthase (mPGES) and cytosolic PGES (cPGES) have recently been identified. In cultured mouse primary osteoblasts, both mPGES and cyclooxygenase 2 were induced by the bone resorptive cytokines interleukin-1, tumor necrosis factor-alpha, and fibroblast growth factor-2. Induction of mPGES was also seen in the mouse long bone and bone marrow in vivo by intraperitoneal injection of lipopolysaccharide. In contrast, cPGES was expressed constitutively both in vitro and in vivo without being affected by these stimuli. An antisense oligonucleotide blocking mPGES expression inhibited not only PGE(2) production, but also osteoclastogenesis and bone resorption stimulated by the cytokines, which was reversed by addition of exogenous PGE(2). We therefore conclude that mPGES, which is induced by and mediates the effects of bone resorptive stimuli, may make a target molecule for the treatment of bone resorptive disorders. PMID- 14566965 TI - TNFalpha suppresses link protein and type II collagen expression in chondrocytes: Role of MEK1/2 and NF-kappaB signaling pathways. AB - Tumor necrosis factor alpha (TNFalpha) inhibits matrix synthesis by chondrocytes in rheumatoid arthritis and osteoarthritis; however, the underlying signaling pathways are poorly characterized. This study investigated the TNFalpha-activated pathways regulating expression of two key components of the cartilage matrix-link protein and type II collagen. In rat articular chondrocytes, TNFalpha decreased link protein and type II collagen mRNA to undetectable levels within 48 h. Levels of link protein mRNA recovered more readily than type II collagen mRNA following removal of the cytokine. TNFalpha-mediated reduction in mRNA of both matrix molecules occurred at the level of transcription and, for link protein, mRNA stability. Turnover of type II collagen and link protein mRNA was dependent on new protein synthesis. In both prechondrocytes and articular chondrocytes, TNFalpha induced concentration-dependent activation of MEK1/2 and NF-kappaB, but not p38 or JNK. Sustained activation of NF-kappaB was observed for up to 72 h following continuous or transient exposure to TNFalpha. Using pharmacological and molecular approaches, the MEK1/2 and NF-kappaB pathways were found to mediate inhibition of type II collagen and link protein gene expression by TNFalpha. Both prechondrocytes and articular chondrocytes are targets of TNFalpha. This study identifies pathways through which TNFalpha perturbs the synthesis and organization of articular cartilage matrix during inflammation. PMID- 14566966 TI - Control of endothelial cell proliferation by calcium influx and arachidonic acid metabolism: a pharmacological approach. AB - In physiological conditions, endothelial cell proliferation is strictly controlled by several growth factors, among which bFGF and VEGF are the most effective. Both bind to specific tyrosine kinase receptors and trigger intracellular signal cascades. In particular, bFGF stimulates the release of arachidonic acid (AA) and its metabolites in many types of endothelial cells in culture. In bovine aortic endothelial cells, it has been suggested that AA is released by the recruitment of cytosolic phospholipase A2 (cPLA2). AA metabolites are involved in the control of both endothelial cell motility (mostly via the cyclooxygenase pathway) and proliferation (via the lipoxygenase (LOX) cascade). On the other hand, evidence has been provided for a proliferative role of AA induced calcium influx. By using a pharmacological approach, we have tried to elucidate the contribution to bovine aortic endothelial proliferation of the different pathways leading to production of AA and its metabolites. Two main informations were obtained by our experiments: first, AA release is not entirely due to cPLA2 involvement, but also to DAG lipase recruitment; second, cyclooxygenase derivatives play a role in the control of cell proliferation, and not only of motility. Moreover, by combining proliferation assays and single cell calcium measurements, we show that the blocking effect of carboxyamido-triazole (CAI), an inhibitor of tumor growth and angiogenesis acting on calcium influx dependent pathways, including AA metabolism, is at least in part due to a direct effect on AA-induced calcium influx. PMID- 14566967 TI - Role of nitric oxide, reactive oxygen species, and p38 MAP kinase in the regulation of human chondrocyte apoptosis. AB - This study addresses mechanisms by which interleukin-1beta (IL-1beta) regulates human chondrocyte apoptosis induced by a combination of the anti-CD95 antibody CH 11 and the proteasome inhibitor (PSI). The effect of IL-1beta on apoptosis varied among tissue samples. IL-1beta either enhanced (16/22 samples) or inhibited (6/22 samples) DNA fragmentation and caspase-3 processing. The protective effect of IL 1beta was abrogated by the nitric oxide (NO) synthesis inhibitor N-monomethyl-l arginine (L-NMMA) while apoptosis stimulation was not affected. The NO-donors sodium nitroprusside (SNP) and S-nitroso-N-acetyl penicillamine (SNAP) blocked DNA fragmentation, and this was associated with partial inhibition of caspase-3 processing. Pyrrolidine dithiocarbamate (PDTC), a scavenger of reactive oxygen species (ROS) blocked apoptosis induction by CH-11/PSI as well as the enhancement by IL-1beta. The pro-apoptotic effects of IL-1beta were also abrogated by the p38 inhibitor SB 202190. In conclusion, IL-1beta augments CH-11/PSI induced apoptosis in the majority of chondrocyte samples. The pro-apoptotic effect of IL-1beta is not dependent on NO. In contrast, the anti-apoptotic effect of IL-1beta observed in a minority of samples is partially NO-dependent. PMID- 14566968 TI - Zinc partitions insulin-like growth factors (IGFs) from soluble IGF binding protein (IGFBP)-5 to the cell surface receptors of BC3H-1 muscle cells. AB - Zinc (Zn(2+)) is a multifunctional micronutrient. The list of functions for this micronutrient expanded with the recent discovery that Zn(2+) retains insulin-like growth factors binding proteins (IGFBPs) on the surface of cultured cells, lowers the affinity of cell-associated IGFBPs, and increases the affinity of the cell surface insulin-like growth factor (IGF)-type 1 receptor (IGF-1R). However, currently there is no information concerning the effect of Zn(2+) on soluble IGFBPs. In the current study, the soluble IGFBP-5 secreted by BC(3)H-1 cells is shown to bind approximately 50% more [(125)I]-IGF-II than [(125)I]-IGF-I at pH 7.4. Zn(2+) is shown to depress the binding of both IGF-I and IGF-II to soluble secreted IGFBP-5; [(125)I]-IGF-I binding is affected more so than [(125)I]-IGF-II binding. Zn(2+) acts by lowering the affinity (K(a)) of IGFBP-5 for the IGFs. Scatchard plots are non-linear indicating the presence of high and low affinity binding sites; Zn(2+) affects only binding to the high affinity site. In contrast, Zn(2+) increases the affinity by which either [(125)I]-IGF-I or [(125)I]-R(3)-IGF-I binds to the IGF-1R, but depresses [(125)I]-IGF-II binding to the IGF-type 2 receptor (IGF-2R) on BC(3)H-1 cells. By depressing the association of the IGFs with soluble IGFBPs, Zn(2+) is shown to repartition either [(125)I] IGF-I or [(125)I]-IGF-II from soluble IGFBP-5 onto cell surface IGF receptors. Zn(2+) was active at physiological doses depressing IGF binding to IGFBP-5 and the IGF-2R at 15-20 microM. Hence, a novel mechanism is further characterized by which the trace micronutrient Zn(2+) could regulate IGF activity. PMID- 14566969 TI - Rab3D redistribution and function in rat parotid acini. AB - Rab3D is a low molecular weight GTP-binding protein believed to be involved with regulated secretion in many cell types. In parotid, Rab3D is localized to secretory granule membranes or present in the cytosol as a complex with Rab escort protein. In the present study, we examined the redistribution of membrane associated Rab3D during secretion in permeabilized parotid acini. When permeabilized acini were stimulated with calcium and cAMP, amylase release increased greater than twofold over basal. Quantitative immunoblotting of subcellular fractions revealed that Rab3D did not dissociate from parotid membranes during secretion. Immunohistochemical staining demonstrated that Rab3D co-localizes with amylase containing granules that are found in the apical pole of the cell. Upon stimulation with calcium and cAMP, Rab3D and amylase immunostaining of granules appeared to be more dispersed. However, Rab3D immunostaining was not observed on the plasma membrane and appeared to reside in the apical cytoplasm. To examine the role of Rab3D in amylase release, cytosolic extracts containing myc-tagged Rab3D and Rab3DQ81L, a GTP-binding mutant, were prepared and incubated with streptolysin O-permeabilized acini. Rab3D, but not Rab3DQ81L, bound to parotid membranes suggesting that Rab3D-binding to parotid membranes is guanine nucleotide-dependent. Moreover, wild-type and mutant Rab3D inhibited agonist-induced amylase release from permeabilized parotid acini. These observations indicate that in parotid acini, Rab3D does not dissociate from parotid membranes or redistribute to the plasma membrane during secretion, and may play an inhibitory role in regulated secretion. The fact that both wild-type Rab3D and the GTP-binding mutant inhibit amylase release suggests that binding of Rab3D to the membrane is not essential for secretory inhibition. PMID- 14566970 TI - Role of Rho in rabbit parietal cell. AB - Rho is known as an important regulator of actin microfilament formation. We were led to study it because a dynamic rearrangement of actin filaments occurs during activation of gastric acid secretion. In order to use specific probes, the rabbit gastric gland culture system was employed and the various genes were expressed using adenovirus vector. When the constitutive active mutant of Rho (RhoAV14) was expressed, histamine- or carbachol-stimulated acid secretion monitored by (14)C aminopyrine accumulation was inhibited. Conversely, expression of C3 toxin, the specific inhibitor of Rho, and expression of G(12/13)-specific regulator of G protein signaling domain, the specific inhibitor of G(12/13) which is considered to be an upstream mediator of Rho, both potentiated acid secretion stimulated by the agonists. F-actin staining of parietal cell expressing RhoAV14 revealed that the microfilament supporting the intracellular canaliculi (not on the basolateral membrane) almost disappeared. No clear changes in the intracellular localization of Rho were observed during stimulation of parietal cell. In resting glands, the endogenous active form of Rho was relatively high, and it decreased during histamine stimulation. The finding that any treatment which inhibit Rho augment acid secretion whereas those that activate Rho inhibit secretion strongly suggests that the Rho-pathway conducts a negatively regulating signal in parietal cell activation, possibly via site-specific regulation of actin microfilaments. PMID- 14566971 TI - Age-related differences in MAP kinase activity in VSMC in response to glucose or TNF-alpha. AB - Aortic vascular smooth muscle cells (VSMC) were used to study the effect of age on responses to high glucose concentrations or the cytokine, tumor necrosis factor-alpha (TNF-alpha). Activator protein-1 (AP-1) binding to DNA increased more in VSMC from old versus young rats (P < 0.02) and was related to increased expression of its components, c-Fos, Fra-1, and JunD. The relationship to upstream signals, i.e., activities of mitogen-activated protein kinases (MAPK), was studied using antibodies to total and phosphorylated forms of extracellular signal-regulated kinases (ERK), c-Jun N-terminal kinases (JNK) and p38. High glucose and TNF-alpha increased ERK phosphorylation more in old (P < 0.05); whereas only TNF-alpha induced JNK activation in young (P < 0.04). PD98059, a MEK inhibitor, attenuated AP-1 activation, lowered c-Fos and Fra-1 protein levels and reduced cell number and cells positive for proliferating cell nuclear antigen in old. We concluded that age differentially influenced activation of signaling pathways in VSMC exposed to high glucose or TNF-alpha. This may contribute to the increased risk for vascular disease associated with aging and diabetes mellitus (DM). PMID- 14566972 TI - JS-K, a novel non-ionic diazeniumdiolate derivative, inhibits Hep 3B hepatoma cell growth and induces c-Jun phosphorylation via multiple MAP kinase pathways. AB - JS-K, a non-ionic diazeniumdiolate derivative, is capable of arylating nucleophiles and spontaneously generating nitric oxide (NO) at physiological pH. This recently synthesized low molecular weight compound is shown here to be an inhibitor of cell growth with concomitant activation of mitogen-activated protein kinase (MAPK) members ERK, JNK, and p38 and their downstream effectors c-Jun and AP-1. Inhibitors of these MAPK pathways abrogated the growth inhibitory actions of JS-K. In addition to the well-described actions of JNK as a kinase for c-Jun, we show that c-Jun is also an ERK target. Furthermore, JS-K generated NO in culture and NO inhibitors antagonized both MAPK induction and the growth inhibitory effects of JS-K. These results suggest two possible mechanisms for the mediation of JS-K growth inhibitory actions, namely NO-induction of MAPK pathway constituents as well as possible arylation reactions. The data support the idea that prolonged MAPK activation by JS-K action is important in mediating its growth-inhibitory actions. JS-K thus represents a promising platform for novel growth inhibitory analog synthesis. PMID- 14566973 TI - Defective P2Y purinergic receptor function: A possible novel mechanism for impaired glucose transport. AB - Extracellular ATP is an ubiquitous mediator that regulates several cellular functions via specific P2 plasma membrane receptors (P2Rs), for which a role in modulating intracellular glucose metabolism has been recently suggested. We have investigated glucose uptake in response to P2Rs stimulation in fibroblasts from type 2 diabetic (T2D) patients and control subjects. P2Rs expression was evaluated by RT-PCR; intracellular calcium release by fluorometry; glucose transporter (GLUT1) translocation by immunoblotting and chemiluminescence; glucose uptake was measured with 2-deoxy-D-[1-(3)H]glucose (2-DOG) and ATP by luminometry. Cells from T2D patients, in contrast to those from healthy controls, showed no increase in glucose uptake after ATP stimulation; extracellular ATP caused, however, a similar GLUT1 recruitment to the plasma membrane in both groups. P2Rs expression did not differ between fibroblasts from diabetic and healthy subjects, but while plasma membrane depolarization, a P2X-mediated response was similar in both groups, no evident intracellular calcium increase was detectable in the cells from the former group. The calcium response in fibroblasts from diabetics was restored by co-incubation with apyrase or hexokinase, suggesting that P2YRs in those cells were normally expressed but chronically desensitised. In support to this finding, fibroblasts from T2D subjects secreted a two-fold larger amount of ATP compared to controls. Pre treatment with apyrase or hexokinase also restored ATP stimulated glucose uptake in fibroblasts from diabetic subjects. These results suggest that extracellular ATP plays a role in the modulation of glucose transport via GLUT1, and that the P2Y-dependent GLUT1 activation is deficient in fibroblasts from T2D individuals. Our observations may point to additional therapeutic targets for improving glucose utilization in diabetes. PMID- 14566974 TI - Stable overexpression of specific segments of the P2P-R protein in human MCF-7 cells promotes camptothecin-induced apoptosis. AB - The stable overexpression of near full-length P2P-R protein in human Saos 2 cells restricts cell cycle progression by inducing mitotic arrest at prometaphase and mitotic apoptosis (Gao and Scott, 2002). Those effects of P2P-R were observed in Saos-2 cells that lack p53 and employ a caspase-3-dependent apoptotic signaling pathway. The current studies were performed to evaluate if overexpression of specific segments of the P2P-R protein promote apoptosis in human MCF-7 cells that contain p53 and employ a different apoptotic signaling pathway. Since segments of P2P-R were found not to induce apoptosis independently, the ability of three different P2P-R segments to promote camptothecin-induced apoptosis was evaluated following their stable transfection and expression in MCF-7 cells. Relative to full-length P2P-R (1-1560 aa), the three P2P-R segments used in these studies included: P2P-R-2 (761-1560 aa), P2P-R-3 (1156-1560 aa), and P2P-R-4 (1314-1560 aa). The results document that overexpression of P2P-R-2 and P2P-R-3 promotes camptothecin-induced apoptosis by three to fivefold when assayed by flow cytometric analysis of apoptotic sub 2n cell populations or by TUNEL assays. In contrast, P2P-R-4 had no effect on apoptosis. These results suggest that the ability of P2P-R to promote camptothecin-induced apoptosis in MCF-7 cells involves a specific region (1156-1314 aa) that exists within P2P-R. The data presented also show that the p53 binding domain of P2P-R overlaps with the apoptosis-associated region and previous studies documented that this region of P2P-R also binds single-strand nucleotides (Witte and Scott, 1997). Therefore, P2P-R-promoted apoptosis induced by camptothecin may be influenced by such interactions. PMID- 14566975 TI - Transforming growth factor-beta induces expression of vascular endothelial growth factor in human retinal pigment epithelial cells: involvement of mitogen activated protein kinases. AB - Vascular endothelial growth factor (VEGF) is a major agent in choroidal and retinal neovascularization, events associated with age-related macular degeneration (AMD) and diabetic retinopathy. Retinal pigment epithelium (RPE), strategically located between retina and choroid, plays a critical role in retinal disorders. We have examined the effects of various growth factors on the expression and secretion of VEGF by human retinal pigment epithelial cell cultures (HRPE). RT-PCR analyses revealed the presence of three isoforms of mRNA corresponding to VEGF 121, 165, and 189 that were up regulated by TGF-beta1. TGF beta1, beta2, and beta3 were the potent inducers of VEGF secretion by HRPE cells whereas bFGF, PDGF, TGF-alpha, and GM-CSF had no effects. TGF-beta receptor type II antibody significantly reversed induction of VEGF secretion by TGF-beta. In contrast activin, inhibin and BMP, members of TGF-beta super family, had no effects on VEGF expression in HRPE. VEGF mRNA levels and protein secretion induced by TGF-beta were significantly inhibited by SB203580 and U0126, inhibitors of MAP kinases, but not by staurosporine and PDTC, protein kinase C and NF-kappaB pathway inhibitors, respectively. TGF-beta also induced VEGF expression by fibroblasts derived from human choroid of eye. TGF-beta induction of VEGF secretion by RPE and choroid cells may play a significant role in choroidal neovascularization (CNV) in AMD. Since the secretion of VEGF by HRPE is regulated by MAP kinase pathways, MAP kinase inhibitors may have potential use as therapeutic agents for CNV in AMD. PMID- 14566976 TI - GFFD: Generalized free-form deformation with scalar fields. AB - The novel free-form deformation(FFD) technique presented in the paper uses scalar fields defined by skeletons with arbitrary topology. The technique embeds objects into the scalar field by assigning a field value to each point of the objects. When the space of the skeleton is changed, the distribution of the scalar field changes accordingly, which implicitly defines a deformation of the space. The generality of skeletons assures that the technique can freely define deformable regions to produce a broader range of shape deformations. PMID- 14566977 TI - LoColms: an innovative approach of enhancing traditional classroom form of education by promoting web-based distance learning in the poorer countries. AB - There have been numerous attempts recently to promote technology based education (Shrestha, 1997) in the poorer third world countries, but so far all these have not provided a sustainable solution as they are either centered and controlled from abroad and relying solely on foreign donors for their sustenance or they are not web-based, which make distribution problematic, and some are not affordable by most of the local population in these places. In this paper we discuss an application, the Local College Learning Management System (LoColms), which we are developing, that is both sustainable and economical to suit the situation in these countries. The application is a web-based system, and aims at improving the traditional form of education by empowering the local universities. Its economy comes from the fact that it is supported by traditional communication technology, the public switching telephone network system, PSTN, which eliminates the need for packet switched or dedicated private virtual networks (PVN) usually required in similar situations. At a later stage, we shall incorporate ontology and paging tools to improve resource sharing and storage optimization in the Proxy Caches (ProCa) and LoColms servers. The system is based on the client/server paradigm and its infrastructure consists of the PSTN, ProCa, with the learning centers accessing the universities by means of point-to-point protocol (PPP). PMID- 14566979 TI - Approach for workflow modeling using pi-calculus. AB - As a variant of process algebra, Pi-calculus can describe the interactions between evolving processes. By modeling activity as a process interacting with other processes through ports, this paper presents a new approach: representing workflow models using Pi-calculus. As a result, the model can characterize the dynamic behaviors of the workflow process in terms of the LTS (Labeled Transition Semantics) semantics of Pi-calculus. The main advantage of the workflow model's formal semantic is that it allows for verification of the model's properties, such as deadlock-free and normal termination. Moreover, the equivalence of workflow models can be checked through weak bisimulation theorem in the Pi calculus, thus facilitating the optimization of business processes. PMID- 14566978 TI - Efficient volume preserving approach for skeleton-based implicit surfaces. AB - This paper presents an efficient way to preserve the volume of implicit surfaces generated by skeletons. Recursive subdivision is used to efficiently calculate the volume. The criterion for subdivision is obtained by using the property of density functions and treating different types of skeletons respectively to get accurate minimum and maximum distances from a cube to a skeleton. Compared with the criterion generated by other ways such as using traditional Interval Analysis, Affine Arithmetic, or Lipschitz condition, our approach is much better both in speed and accuracy. PMID- 14566980 TI - Optimization of block-floating-point realizations for digital controllers with finite-word-length considerations. AB - The closed-loop stability issue of finite-precision realizations was investigated for digital controllers implemented in block-floating-point format. The controller coefficient perturbation was analyzed resulting from using finite word length (FWL) block-floating-point representation scheme. A block-floating-point FWL closed-loop stability measure was derived which considers both the dynamic range and precision. To facilitate the design of optimal finite-precision controller realizations, a computationally tractable block-floating-point FWL closed-loop stability measure was then introduced and the method of computing the value of this measure for a given controller realization was developed. The optimal controller realization is defined as the solution that maximizes the corresponding measure, and a numerical optimization approach was adopted to solve the resulting optimal realization problem. A numerical example was used to illustrate the design procedure and to compare the optimal controller realization with the initial realization. PMID- 14566981 TI - Applications of cascade multilevel inverters. AB - Cascade multilevel inverters have been developed for electric utility applications. A cascade M-level inverter consists of (M-1)/2 H-bridges in which each bridge's dc voltage is supported by its own dc capacitor. The new inverter can: (1) generate almost sinusoidal waveform voltage while only switching one time per fundamental cycle; (2) dispense with multi-pulse inverters' transformers used in conventional utility interfaces and static var compensators; (3) enables direct parallel or series transformer-less connection to medium- and high-voltage power systems. In short, the cascade inverter is much more efficient and suitable for utility applications than traditional multi-pulse and pulse width modulation (PWM) inverters. The authors have experimentally demonstrated the superiority of the new inverter for power supply, (hybrid) electric vehicle (EV) motor drive, reactive power (var) and harmonic compensation. This paper summarizes the features, feasibility, and control schemes of the cascade inverter for utility applications including utility interface of renewable energy, voltage regulation, var compensation, and harmonic filtering in power systems. Analytical, simulated, and experimental results demonstrated the superiority of the new inverters. PMID- 14566982 TI - An electricity price model with consideration to load and gas price effects. AB - Some characteristics of the electricity load and prices are studied, and the relationship between electricity prices and gas (fuel) prices is analyzed in this paper. Because electricity prices are strongly dependent on load and gas prices, the authors constructed a model for electricity prices based on the effects of these two factors; and used the Geometric Mean Reversion Brownian Motion (GMRBM) model to describe the electricity load process, and a Geometric Brownian Motion(GBM) model to describe the gas prices; deduced the price stochastic process model based on the above load model and gas price model. This paper also presents methods for parameters estimation, and proposes some methods to solve the model. PMID- 14566983 TI - Analysis of tensioned membrane structures considering cable sliding. AB - In routine design of tensioned membrane structures, the membrane is generally modeled using space membrane elements and the cables by space cable elements, with no sliding allowed between the membrane and the cables. On the other hand, large deflections are expected and sliding between the membrane and the cables is inevitable. In the present paper, the general finite element code ABAQUS was employed to investigate the influence of cable sliding on membrane surface on the structural behavior. Three analysis models were devised to fulfill this purpose: (1) The membrane element shares nodes with the cable element; (2) The cable can slide on the membrane surface freely (without friction) and (3) The cable can slide on the membrane surface, but with friction between the cable and the membrane. The sliding problem is modeled using a surface-based contact algorithm. The results from three analysis models are compared, showing that cable sliding has only little influence on the structure shape and on the stress distributions in the membrane. The main influence of cable sliding may be its effect on the dynamic behavior of tensioned membrane structures. PMID- 14566984 TI - Study on pile drivability with one dimensional wave propagation theory. AB - Pile drivability is a key problem during the stage of design and construction installation of pile foundations. The solution to the one dimensional wave equation was used to determine the impact force at the top of a concrete pile for a given ram mass, cushion stiffness, and pile impedance. The kinematic equation of pile toe was established and solved based on wave equation theory. The movements of the pile top and pile toe were presented, which clearly showed the dynamic displacement, including rebound and penetration of pile top and toe. A parametric study was made with a full range of practical values of ram weight, cushion stiffness, drop eight, and pile impedance. Suggestions for optimizing the parameters were also presented. Comparisons between the results obtained by the present solution and in-situ measurements indicated the reliability and validity of the method. PMID- 14566986 TI - Life cycle assessment in the environmental impact evaluation of urban development -a case study of land readjustment project, Hyogo District, Japan. AB - In this paper, the Life Cycle of Urban Development was firstly analyzed, and the phases of Life Cycle Assessment applied to Urban Development (ULCA) were described. As a case study, ULCA was applied in the environmental impact assessment of the land readjustment project of Hyogo District of Saga, Japan. In addition, mitigation proposals for reducing CO(2) were also presented and the relevant environmental effects were simulated. PMID- 14566985 TI - Influence of water saturation on propagation of elastic waves in transversely isotropic nearly saturated soil. AB - Biot's two-phase theory for fluid-saturated porous media was applied in a study carried out to investigate the influence of water saturation on propagation of elastic wave in transversely isotropic nearly saturated soil. The characteristic equations for wave propagation were derived and solved analytically. The results showed that there are four waves: the first and second quasi-longitudinal waves (QP1 and QP2), the quasi-transverse wave (QSV) and the anti-plane transverse wave (SH). Numerical results are given to illustrate the influence of saturation on the velocity, dispersion and attenuation of the four body waves. Some typical numerical results are discussed and plotted. The results can be meaningful for soil dynamics and earthquake engineering. PMID- 14566987 TI - A method for predicting critical load evaluating adhesion of coatings in scratch testing. AB - In this paper based on the experiment principle of evaluating adhesion property by scratch testing, the peeling mechanism of thin films is discussed by applying contact theory and surface physics theory. A mathematical model predicting the critical load is proposed for calculating critical load as determined by scratch testing. The factors for correctly evaluating adhesion of coatings according to the experimental data are discussed. PMID- 14566988 TI - A novel long-chain acyl-derivative of epigallocatechin-3-O-gallate prepared and purified from green tea polyphenols. AB - Lipophilic tea polyphenols (LTP) were prepared by catalytic esterification of green tea polyphenols (GTP) with hexadecanoyl chloride. A novel long-chain acyl derivative of epigallocatechin-3-O-gallate (EGCG) was first isolated from purification of LTP by high-speed countercurrent chromatography (HSCCC) using a solvent system composed of n-hexane-ethyl acetate-methanol-water (1:1:1:1, v/v). The molecular structure of the acyl-derivative, Epigallocatechin-3-O-gallate-4'-O hexadecanate, was elucidated by means of elemental analysis, IR, (1)H-NMR and MS spectra PMID- 14566989 TI - Optimization of cultural conditions for thermostable beta-1,3-1,4-glucanase production by Bacillus subtilis ZJF-1A5. AB - The optimization of cultural conditions for Beta-glucanase production by Bacillus subtilis ZJF-1A5 was investigated in flask trials. Temperature had great effect on Beta-glucanase production which maximized at optimal temperature of 37 degrees C and decreased significantly when temperature was over 37 degrees C. Charge quantity affected Beta-glucanase production significantly. Adding oxygen vector N dodecane or acetic ether benefited Beta-glucanase production, but it depended on the concentration and charge quantity. The results of fractional factorial design showed that age and size of inoculum and shaking speed were the key factors affecting Beta-glucanase production and the cultivation time span to reach the highest Beta-glucanase activity. The optimal cultural conditions for Beta glucanase production obtained with CCD were as follows: inoculum age and size (16 h, 3.82%(v/v)), shaking speed 210 r/min, charge quantity of 30 mL in 250 mL flask and initial pH 7.0, cultured at 37 degrees C for 50 h. Repeated experimental results accorded with those predicted by a second-order polynomial model. The amount of Beta-glucanase, Alpha-amylase and neutral protease produced by B subtilis ZJF-1A5 was associated partially with cell growth. Those three enzymes' activities increased following the cell growth and increased significantly when cells entered the stationary phase. PMID- 14566990 TI - In situ hyperspectral data analysis for pigment content estimation of rice leaves. AB - Analyses of the correlation between hyperspectral reflectance and pigment content including chlorophyll-a, chlorophyll-b and carotenoid of leaves in different sites of rice were reported in this paper. The hyperspectral reflectance of late rice during the whole growing season was measured using a Spectroradiometer with spectral range of 350-1050 nm and resolution of 3 nm. The chlorophyll-a, chlorophyll-b and carotenoid contents in rice leaves in rice fields to which different levels of nitrogen were applied were measured. The chlorophyll-a content of upper leaves was well correlated with the spectral variables. However, the correlation between both chlorophyll-b and caroteniod and the spectral variables was far from that of chlorophyll-a. The potential of hyperspectral reflectance measurement for estimating chlorophyll-a of upper leaves was evaluated using univariate correlation and multivariate regression analysis methods with different types of predictors. This study showed that the most suitable estimated model of chlorophyll-a of upper leaves was obtained by using some hyperspectral variables such as SD(r), SD(b) and their integration. PMID- 14566991 TI - Study on pig growth hormone gene polymorphisms in western meat-type breeds and Chinese local breeds. AB - Chinese Meishan and Jiangquhai pigs are two of the most prolific pigs in the world, but their growth rate is lower than that of Duroc, Landrace and Pietrain pigs. It is suggested that growth rate is regulated by growth hormone. The objective of the current study was to analyze the porcine growth hormone (pGH) gene polymorphisms based on the polymerase chain reaction restriction fragment length polymorphism method (PCR-RFLP) for three western meat-type breeds (Duroc, Landrace and Pietrain) and two local Chinese pigs (Meishan and Jiangquhai). Five polymorphic restriction sites were detected with the ApaI, MspI, BspI and HhaI restriction enzymes in two amplified fragments (605 bp, -119 to +486; 506 bp, +206 to +711). Breed difference was found only in the 506 bp fragment. There was no difference in allelic frequencies of BspI and HhaI restriction sites among the five breeds (P>0.05). Landrace and Meishan pigs lacked allele G3 of MspI site. The allele G3 frequency of restriction MspI site of the 506 bp fragment in Pietrain pigs was higher than that in Duroc and Jianquhai pigs (P<0.001). For ApaI site, the Meishan pigs lacked allele G1; no difference was found in allelic frequencies among Pietrain, Duroc, Landrace and Jiangquhai pigs (P>0.05). This new and rapid PCR-RFLP typing method is an attractive tool for analysis of porcine growth hormone gene restriction sites. The differences in MspI and ApaI restriction sites may explain the growth difference between the foreign meat-type breeds above mentioned and local Chinese pigs. PMID- 14566992 TI - On the p-norm joint spectral radius. AB - The p-norm joint spectral radius is defined by a bounded collection of square matrices with complex entries and of the same size. In the present paper the author investigates the p-norm joint spectral radius for integers. The method introduced in this paper yields some basic formulas for these spectral radii. The approach used in this paper provides a simple proof of Berger-Wang's relation concerning the infinity -norm joint spectral radius. PMID- 14566993 TI - Ultrasound ablation of pulmonary veins for treatment of paroxysmal atrial fibrillation. AB - OBJECTIVE: to evaluate the efficacy and safety of ultrasound ablation of pulmonary veins for treatment of paroxysmal atrial fibrillation. METHODS: The study population consisted of 9 patients with 5 males and 4 females enrolled consecutively who had idiopathic paroxysmal atrial fibrillation(AF). The ultrasound balloon was positioned through a special sheath to the orifice of the target vein by a transseptal procedure. The balloon was inflated with contrast mixed saline (contrast:saline = 1:4) whose volume was decided by the diameter of the target pulmonary vein. The ablation energy was usually set up at 35 to 40 watts with temperature controlled at 60 degrees C. The duration of each ablation was about 120 seconds which was repeated not over 10 times. RESULTS: The average duration of the total procedure was 132+/-68 min for our patients. The average fluoroscopy time was 33+/-17 min. With a mean follow-up of 16+/-8 months after the procedure, AF was completely eliminated in 4 patients without antiarrhythmic drugs. The episodes of atrial fibrillation were eliminated in 2 patients with low dosage of oral amiodarone (0.1, once daily) which was ineffective before the procedure. The frequency of episodes was similar to that before the procedure in 3 patients. There were no complications such as hemopericardium, air embolism and stenosis of the pulmonary veins by angiography, related to the procedure. CONCLUSION: Ultrasound ablation of the pulmonary veins is a new approach to treat paroxysmal atrial fibrillation. Before we determined its value, we need to do more researches with bigger sample, randomization and comparison design. PMID- 14566994 TI - Study of apolipoprotein E genetic polymorphism in patients with atherosclerotic cerebral infarction. AB - OBJECTIVE: To explore the frequency and significance of ApoE gene polymorphisms in Chinese patients with atherosclerotic cerebral infarction (ACI). METHODS: Polymerase chain reaction and gene sequencing, single nucleotide polymorphisms of ApoE gene were used to analyze 33 cases of patients with ACI and 35 controls. RESULTS: The frequencies of ApoE gene single nucleotide polymorphisms 465C/G, 462C/G and 451delC in the ACI group were significantly higher than those in the control group (P<0.05). The prevalence of polymorphism 486G/T in the control group was significantly higher than that in the ACI group (P=0.011). CONCLUSIONS: 465C/G,462C/G and 451delC polymorphisms might be associated with ACI.486GT allele might have protective effect on the pathogenesis of ACI. PMID- 14566995 TI - Effects of breakfast with different calorigenic amounts on blood glucose, insulin and glucagon levels. AB - This study was aimed to investigate the relationship between breakfast and serum glucose, insulin and glucagon concentrations in order to establish a model breakfast appropriate for Chinese. Twenty-four volunteers were randomly assigned to four study groups: high carbohydrate breakfast, high fat and protein breakfast, the typical breakfast and fasting. Each subject had serum and urine samples collected while fasting and at 1,2 and 3.5 hours following the meal. The concentration of serum glucose, insulin and glucagon was measured. The levels of serum glucose in group A, B and C differed significantly at 1 and 2 hour after meal compared to those at fasting (P<0.05). The serum glucose in group A increased insignificantly after meal. The serum insulin levels were in group A, B and C significant different compared with control group(P<0.05). Those peaked at 1 hour after meal, with group C rising the furthest. Compared with the fasting group, the serum glucagons rose and maintained the increase after breakfast in group A, B and C (P<0.05). The data suggested that various diets with different calorigenic amounts increased hormone concentration to various extents. We found that a breakfast rich in carbohydrates could maintain proper blood glucose level. PMID- 14566996 TI - Pediatric respiratory tract infections: etiology, microbiology and resistance trends. Introduction. PMID- 14566997 TI - Worldwide trends in antimicrobial resistance among common respiratory tract pathogens in children. AB - BACKGROUND: Respiratory tract infections among children are a common reason for health care provider visits and the primary reason for antimicrobial prescribing in this population. The increased prevalence of resistance among Streptococcus pneumoniae and Haemophilus influenzae pathogens poses a serious challenge in the successful treatment of respiratory tract infections caused by these pathogens. METHODS: This paper reviews worldwide trends in antimicrobial resistance among common respiratory tract pathogens, highlighting data obtained from the pediatric population where available. RESULTS: S. pneumoniae resistance to beta-lactams is mediated through alterations in the penicillin-binding proteins and macrolide resistance to acquisition of efflux or methylation genes. The mechanisms of resistance to the fluoroquinolones include target enzyme alterations via genetic mutations and transport out of the bacterial cell via an efflux pump. Beta lactamase production is the primary mechanism of resistance to penicillins among H. influenzae isolates. Although S. pneumoniae with reduced susceptibility to penicillin was first documented > 30 years ago, resistance has increased at an alarming rate worldwide in the past decade. According to recent surveillance data, the worldwide prevalence of S. pneumoniae with reduced susceptibility to penicillin is 18.2%. Beta-lactamase production among H. influenzae ranges from approximately 4% in Russia to 26% in the United States and to 31% in France. The prevalence of beta-lactamase-negative, ampicillin-resistant H. influenzae remains very low (< 1%) worldwide, except in Japan, where the incidence is higher. In general, the highest rates of resistance are observed in isolates obtained from children, and risk factors for infection with a resistant pathogen include young age, the site of infection, day-care center attendance and recent antimicrobial use. CONCLUSIONS: Increased prevalence of antimicrobial resistance among respiratory tract pathogens isolated from children and adults is evident worldwide. Treatment of infections caused by S. pneumoniae and H. influenzae with older agents or ineffective dosing regimens may not eradicate infections and may contribute to the spread of resistance. These observations confirm the need for appropriate antimicrobial use to halt or at least limit the spread of resistance.